PMID- 12527357 TI - Photoreceptor synaptic protein HRG4 (UNC119) interacts with ARL2 via a putative conserved domain. AB - Human retinal gene 4 (HRG4) (UNC119) is a photoreceptor synaptic protein of unknown function, shown when mutated to cause retinal degeneration in a patient and in a confirmatory transgenic model. ADP-ribosylation factor-like protein 2 (ARL2) was identified as an interactor of HRG4 by the yeast two-hybrid strategy. The presence of ARL2 in the retina and co-localization with HRG4 was confirmed by Western blot and double immunofluorescence analysis, respectively. The interaction of ARL2 with HRG4 was further confirmed by co-immunoprecipitation and direct binding analysis. Phosphodiesterase delta (PDEdelta) is an ARL2-binding protein homologous to HRG4. Amino acid residues of PDEdelta involved in binding ARL2 and forming a hydrophobic pocket were shown to be highly conserved in HRG4, suggesting similarity in binding mechanism and function. PMID- 12527358 TI - Induction of neuron-like tubes and liposome networks by cooperative effect of gangliosides and phospholipids. AB - Although there is a rather large abundance of gangliosides in neurons, their functional role is still unclear. We focused on a physicochemical role of gangliosides in the formation of tubular structures, such as axons or dendrites in neurons. When a ganglioside, GM3, was added to cell-size liposomes that consisted of dioleoylphosphatidyl-choline, tubular structures were induced and liposome networks connected by the tubes were observed by differential interference microscopy and fluorescence microscopy. The potential for various gangliosides to induce tubes was dependent on the structures of their hydrophilic head group. With a large excess of gangliosides, the tubes are destabilized and small fragments, or micelles, are generated. The phenomenon was suggested by physical model calculation. Gangliosides may play a role as building material in neural unique tubular structures. PMID- 12527359 TI - The complete mitochondrial genome sequence of the pathogenic yeast Candida (Torulopsis) glabrata. AB - We report here the complete sequence of the mitochondrial (mt) genome of the pathogenic yeast Candida glabrata. This 20 kb mt genome is the smallest among sequenced hemiascomycetous yeasts. Despite its compaction, the mt genome contains the genes encoding the apocytochrome b (COB), three subunits of ATP synthetase (ATP6, 8 and 9), three subunits of cytochrome oxidase (COX1, 2 and 3), the ribosomal protein VAR1, 23 tRNAs, small and large ribosomal RNAs and the RNA subunit of RNase P. Three group I introns each with an intronic open reading frame are present in the COX1 gene. This sequence is available under accession number AJ511533. PMID- 12527360 TI - The enzymatic activity of phosphoglucose isomerase is not required for its cytokine function. AB - PGI is a housekeeping gene encoding phosphoglucose isomerase (PGI) a glycolytic enzyme that also functions as a cytokine (autocrine motility factor (AMF)/neuroleukin/maturation factor) upon secretion from the cell and binding to its 78 kDa seven-transmembrane domain receptor (gp78/AMF-R). PGI contains a CXXC motif, characteristic of redox proteins and possibly evolutionarily related to the CC and CXC motif of the chemokine gene family. Using site-directed mutagenesis, single- and double-deletion (CXC, CC) mutants were created by deleting amino acids 331 and 332 of human PGI, respectively. The mutant proteins lost their enzymatic activity; however, neither of the deletions augmented the proteins' binding affinity to the receptor and all maintained cytokine function. The results demonstrate that the enzymatic activity of PGI is not essential for either receptor binding or cytokine function of human PGI. PMID- 12527361 TI - Glucosamine-induced insulin resistance is coupled to O-linked glycosylation of Munc18c. AB - Evidence suggests that glucosamine inhibits distal components regulating insulin stimulated GLUT4 translocation to the plasma membrane. Here we assessed whether key membrane docking and fusion events were targeted. Consistent with a plasma membrane-localized effect, 3T3-L1 adipocytes exposed to glucosamine displayed an increase in cell-surface O-linked glycosylation and a simultaneously impaired mobilization of GLUT4 by insulin. Analysis of syntaxin 4 and SNAP23, plasma membrane-localized target receptor proteins (t-SNAREs) for the GLUT4 vesicle, showed that they were not cell-surface targets of O-linked glycosylation. However, the syntaxin 4 binding protein, Munc18c, was targeted by O-linked glycosylation. This occurred concomitantly with a block in insulin-stimulated association of syntaxin 4 with its cognate GLUT4 vesicle receptor protein (v SNARE), VAMP2. In conclusion, our data suggest that the mechanism by which glucosamine inhibits insulin-stimulated GLUT4 translocation involves modification of Munc18c. PMID- 12527362 TI - Bcl-rambo beta, a special splicing variant with an insertion of an Alu-like cassette, promotes etoposide- and Taxol-induced cell death. AB - The exonization of an Alu-like element into a coding sequence is unique to primates and this phenomenon distinguishes our genome from other mammals. Here, we report the presence of a special splicing variant of a proapoptotic protein Bcl-rambo in human lymph node, designated as Bcl-rambo beta. This variant contains a 98 bp Alu-like sequence which acts as an exon. There exists an in frame stop codon within this inserted Alu-like cassette, resulting in generation of a premature protein of 104 amino acid residues. Unlike the Bcl-rambo, Bcl rambo beta is lacking of the BH1, BH2 and BH3 motifs and becomes a BH4-only protein. Bcl-rambo beta is detected in several adult human tissues such as heart, lymph node and cervix but is absent in human brain tissue. In addition, Bcl-rambo beta is found not to be associated with mitochondria due to the absence of its C terminal membrane anchor region. Nevertheless, this cytosol-localized protein is capable of promoting etoposide- and Taxol-induced cell death. Although the exact function of the Alu sequence is not fully characterized, the Alu element within the Bcl-rambo beta appeared to contribute to the proapoptotic capability, since removing of the Alu sequence from Bcl-rambo beta abrogates its ability to induce cell death. Our data support the speculation that the Alu element insertion during the splicing process may play an important role in the generation of protein diversity in primate cells by a yet uncharacterized mechanism. PMID- 12527363 TI - Differential phosphorylation activities of CDK-activating kinases in Arabidopsis thaliana. AB - Activation of cyclin-dependent kinases (CDKs) requires phosphorylation of a threonine residue within the T-loop by a CDK-activating kinase (CAK). Here we isolated an Arabidopsis cDNA (CAK4At) whose predicted product shows a high similarity to vertebrate CDK7/p40(MO15). Northern blot analysis showed that expressions of the four Arabidopsis CAKs (CAK1At-CAK4At) were not dependent on cell division. CAK2At- and CAK4At-immunoprecipitates of Arabidopsis crude extract phosphorylated CDK and the carboxy-terminal domain (CTD) of the largest subunit of RNA polymerase II with different preferences. These results suggest the existence of differential mechanisms in Arabidopsis that control CDK and CTD phosphorylation by multiple CAKs. PMID- 12527364 TI - Isolation of a new dual-functional caffeine synthase gene encoding an enzyme for the conversion of 7-methylxanthine to caffeine from coffee (Coffea arabica L.). AB - In coffee and tea plants, caffeine is synthesized from xanthosine via a pathway that includes three methylation steps. We report the isolation of a bifunctional coffee caffeine synthase (CCS1) clone from coffee endosperm by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) technique using previously reported sequence information for theobromine synthases (CTSs). The predicted amino acid sequences of CCS1 are more than 80% identical to CTSs and are about 40% similar to those of tea caffeine synthase (TCS1). Interestingly, CCS1 has dual methylation activity like tea TCS1. PMID- 12527365 TI - A tomato xylem sap protein represents a new family of small cysteine-rich proteins with structural similarity to lipid transfer proteins. AB - The coding sequence of a major xylem sap protein of tomato was identified with the aid of mass spectrometry. The protein, XSP10, represents a novel family of extracellular plant proteins with structural similarity to plant lipid transfer proteins. The XSP10 gene is constitutively expressed in roots and lower stems. The decline of XSP10 protein levels in tomato infected with a fungal vascular pathogen may reflect breakdown or modification by the pathogen. PMID- 12527366 TI - Identification of an Arabidopsis mitochondrial succinate-fumarate translocator. AB - Complementation of a yeast acr1 mutant carrying a deletion of the succinate/fumarate carrier gene enabled functional identification of a mitochondrial succinate translocator from Arabidopsis thaliana (AtmSFC1). Thus complementation of yeast mutants is applicable also for identification and characterization of organellar transporters. Reverse transcription polymerase chain reaction and promoter-GUS fusion showed expression of AtmSFC1 in 2 day old dark grown seedlings, which declined in cotyledons during further development, consistent with a role in export of fumarate for gluconeogenesis during lipid mobilization at early germination of Arabidopsis seeds. In mature plants, expression was found in developing and germinating pollen, suggesting a role in ethanolic fermentation. PMID- 12527367 TI - A novel zinc finger transcriptional repressor, ZNF224, interacts with the negative regulatory element (AldA-NRE) and inhibits gene expression. AB - The interaction between the negative cis-element (AldA-NRE) and p97 repressor nuclear protein is a key step in modulating transcription of the human and mouse aldolase A (AldA) gene during the cell cycle and differentiation. In an attempt to clarify the role of transcriptional repression in regulating gene expression, we purified, from HeLa cells, the nuclear protein that specifically binds to the AldA negative regulatory element (NRE). Matrix-assisted laser desorption ionization-time of flight analysis and examination of protein profiles from the SwissProt database revealed that the previously defined p97 repressor is ZNF224, a zinc finger protein. We demonstrate that ZNF224, a Kruppel-like zinc finger transcription factor, is the repressor protein that specifically binds to the negative cis-element AldA-NRE and affects the AldA-NRE-mediated transcription. PMID- 12527368 TI - Ca2+ oscillations in hepatocytes do not require the modulation of InsP3 3-kinase activity by Ca2+. AB - Receptor-mediated production of inositol 1,4,5-trisphosphate (InsP(3)) initiates Ca(2+) release and is responsible for cytosolic Ca(2+) oscillations. InsP(3) oscillations have also been observed in some cells. One of the enzymes controlling InsP(3) catabolism, the InsP(3) 3-kinase, is stimulated by Ca(2+); this regulation is presumably part of the reason for InsP(3) oscillations that have been observed in some cells. Here, we investigate the possible role of Ca(2+)-activated InsP(3) catabolism on the characteristics of the InsP(3)-induced Ca(2+) oscillations. Numerical simulations show that if it is assumed that the Ca(2+)-independent InsP(3) catabolism is predominant, Ca(2+) oscillations remain qualitatively unchanged although the relative amplitude of the oscillations in InsP(3) concentrations becomes minimal. We tested this prediction in hepatocytes by masking the Ca(2+)-dependent InsP(3) catabolism by 3-kinase through the injection of massive amounts of InsP(3) 5-phosphatase, which is not stimulated by Ca(2+). We find that in such injected hepatocytes, Ca(2+) oscillations generated by modest agonist levels are suppressed, presumably because of the decreased dose in InsP(3), but that at higher doses of agonist, oscillations reappear, with characteristics similar to those of untreated cells at low agonist doses. Altogether, these results suggest that oscillations in InsP(3) concentration due to Ca(2+)-stimulated InsP(3) catabolism do not play a major role for the oscillations in Ca(2+) concentration. PMID- 12527369 TI - Self-aggregation of legume seed storage proteins inside the protein storage vacuoles is electrostatic in nature, rather than lectin-mediated. AB - Conglutins are multisubunit, glycosylated, major storage proteins present in Lupinus seeds that self-aggregate in a calcium/magnesium-dependent manner. Two of these globulins exhibit lectin activity. The 210 kDa globulin derived from beta conglutin that accumulates in Lupinus cotyledons during germination was used as a model protein to establish whether the self-aggregation process is electrostatic in nature or lectin-mediated. This protein binds in a very strong manner to chitin and recognizes a variety of glycoproteins including immunoglobulins G. Several compounds were tested for their inhibitory effect on the cation-dependent self-aggregation process. Sialic acid and phytin were the most effective whereas chitin and mucin were totally ineffective. The inability of the oligosaccharidic side chains of the 210 kDa protein, beta-conglutin and immunoglobulin G to interfere with the aggregation strongly supports the view that Ca/Mg are electrostatically involved in the in vitro self-aggregation of Lupinus globulins. The results suggest that calcium and magnesium ions are also electrostatically involved in vivo in the macromolecular aggregation of legume seed storage proteins, ensuring their efficient packing inside the protein storage vacuoles. This mechanism is responsible for the typical insolubility of legume globulins in water. PMID- 12527370 TI - Halving the selenophosphate synthetase gene dose confers hypersensitivity to oxidative stress in Drosophila melanogaster. AB - Several lines of evidence indicate that selenoproteins mainly act as cellular antioxidants. Here, we test this idea comparing the sensitivity to oxidative stress (paraquat and hydrogen peroxide) between wild type and heterozygous flies for the selenophosphate synthetase selD(ptuf) mutation. Whereas under normal laboratory conditions no difference in life span is observed, a significant decrease is seen in heterozygous flies treated with oxidant agents. In contrast, overexpression of the selD gene in motoneurons did not extend longevity. Our results strongly suggest that selD haploinsufficiency makes heterozygous flies more sensitive to oxidative stress and add further evidence to the role of selenoproteins as cellular antioxidants. PMID- 12527371 TI - Involvement of acetylated tubulin in the regulation of Na+,K+ -ATPase activity in cultured astrocytes. AB - The results presented support the view that the modulation of Na(+),K(+)-ATPase activity in living cells involves the association/dissociation of acetylated tubulin with the enzyme. We found that the stimulation of Na(+),K(+)-ATPase activity by L-glutamate correlates with decreased acetylated tubulin quantity associated with the enzyme. The effect of L-glutamate was abolished by the glutamate transporter inhibitor DL-threo-beta-hydroxyaspartate but was not affected by either specific agonists or antagonists. The effect of L-glutamate seems to be mediated by Na(+) entry resulting from glutamate transport, since the Na(+) ionophore monensin produced stimulation of Na(+),K(+)-ATPase activity with concomitant decrease of acetylated tubulin quantity associated with the enzyme. PMID- 12527372 TI - Myb induced myeloid protein 1 (Mim-1) is an acetyltransferase. AB - We have screened protein extracts from chicken blood cells for acetyltransferases. An in gel acetyltransferase assay revealed that a 32 kDa protein, which is more prevalent in whole blood when compared with erythrocyte cells, possessed an auto-acetylation activity. This protein was purified by a series of chromatographic steps, sequenced by Edman degradation and subsequently identified as Myb induced myeloid protein (Mim-1). Mim-1 has similarities to the conserved acetyltransferase motifs found in the GNAT superfamily of proteins and also contains three minimal GK acetylation motifs. These data identify Mim-1 as an acetyltransferase. PMID- 12527373 TI - Normal function of HERG K+ channels expressed in HEK293 cells requires basal protein kinase B activity. AB - The potential role of protein kinase B (PKB), a serine/threonine protein kinase, in regulating HERG (human ether-a-go-go related gene) K(+) channel function was investigated. Wortmannin (a phosphoinositide 3-kinase (PI3K) inhibitor) caused approximately 30% reduction of HERG current (I(HERG)) stably expressed in HEK293 cells. Transient transfection with the constitutively active PI3K in HERG expressing HEK293 cells slightly increased ( approximately 7%) I(HERG) while a dominant negative PI3K significantly reduced I(HERG) ( approximately 25%) relative to results in vehicle-transfected cells. I(HERG) was approximately 35% greater in cells transfected with the constitutively activated PKB (caPKB), whereas it was approximately 47% smaller in cells transfected with dominant negative PKB (dnPKB). Basal activation of PKB was detected by immunocytochemistry. PKB activity was significantly enhanced in caPKB-transfected cells and nearly abolished in dnPKB-transfected cells. We conclude that normal HERG function in HEK293 cells requires basal activity of PKB. Our data represent the first evidence that PKB phosphorylation regulates K(+) channels. PMID- 12527374 TI - Distinct roles of the Src family kinases, SRC-1 and KIN-22, that are negatively regulated by CSK-1 in C. elegans. AB - To elucidate the primitive roles of the Src family kinases (SFKs), here we characterized Caenorhabditis elegans orthologues of SFKs (src-1 and kin-22) and their regulator kinase Csk (csk-1). SRC-1 and KIN-22 possess the C-terminal regulatory tyrosines characteristic of SFKs, and their activities are negatively regulated by CSK-1 in a yeast expression system. The src-1 and csk-1 genes are co expressed in some head neurons, the anchor cell and the tail region, while kin-22 and csk-1 genes are co-expressed in pharyngeal muscles and tail region. Expression of KIN-22 induced morphological defects in the pharynx, whereas expression of SRC-1 did not show any overt phenotype in adult. RNA interference of src-1, but not that of kin-22, caused a developmental arrest in early development. These results suggest that SRC-1 and KIN-22 play distinct roles under the control of CSK-1. PMID- 12527375 TI - The processing of human mitochondrial leucyl-tRNA synthetase in the insect cells. AB - A His-tagged full-length cDNA of human mitochondrial leucyl-tRNA synthetase was expressed in a baculovirus system. The N-terminal sequence of the enzyme isolated from the mitochondria of insect cells was found to be IYSATGKWTKEYTL, indicating that the mitochondrial targeting signal peptide was cleaved between Ser39 and Ile40 after the enzyme precursor was translocated into mitochondria. The enzyme purified from mitochondria catalyzed the leucylation of Escherichia coli tRNA(1)(Leu)(CAG) and Aquifex aeolicus tRNA(Leu)(GAG) with higher catalytic activity in the leucylation of E. coli tRNA(Leu) than that previously expressed in E. coli without the N-terminal 21 residues. PMID- 12527376 TI - Pseudomonas stutzeri soluble nitrate reductase alphabeta-subunit is a soluble enzyme with a similar electronic structure at the active site as the inner membrane-bound alphabetagamma holoenzyme. AB - A two-subunit (alphabeta) form of dissimilatory nitrate reductase from Pseudomonas stutzeri strain ZoBell was separated from the membrane-residing gamma subunit by a heat solubilization step. Here we present an optimized purification protocol leading to a soluble alphabeta form with high specific activity (70 U/mg). The soluble form has the stoichiometry alpha(1)beta(1) consisting of the 130 kDa alpha-subunit and the 58 kDa beta-subunit. We did not observe any proteolytic cleavage in the course of the heat solubilization. The enzyme is competively inhibited by azide, but not by chlorate. It exhibits a K(M) value of 3.2 mM for nitrate. We compare the enzymatic and electron paramagnetic resonance (EPR) spectroscopic properties of the alphabeta form with the alphabetagamma holoenzyme which resides in the membrane and can be prepared by detergent extraction. The nearly identical EPR spectra for the Mo(V) signal of both enzyme preparations show that the active site is unaffected by the heat step. The factors influencing the binding of the alpha- and beta-subunit to the gamma subunit are discussed. PMID- 12527377 TI - A motif rich in charged residues determines product specificity in isomaltulose synthase. AB - Isomaltulose synthase (PalI) catalyzes hydrolysis of sucrose and formation of alpha-1,6 and alpha-1,1 bonds to produce isomaltulose (alpha-D-glucosylpyranosyl 1,6-D-fructofranose) and small amount of trehalulose (alpha-D-glucosylpyranosyl 1,1-D-fructofranose). A potential isomaltulose synthase-specific motif ((325)RLDRD(329)), that contains a 'DxD' motif conserved in many glycosyltransferases, was identified based on sequence comparison with reference to the secondary structural features of PalI and homologs. Site-directed mutagenesis analysis of the motif showed that the four charged amino acid residues (Arg(325), Arg(328), Asp(327) and Asp(329)) influence the enzyme kinetics and determine the product specificity. Mutation of these four residues increased trehalulose formation by 17-61% and decreased isomaltulose by 26-67%. We conclude that the 'RLDRD' motif controls the product specificity of PalI. PMID- 12527378 TI - DmsD is required for the biogenesis of DMSO reductase in Escherichia coli but not for the interaction of the DmsA signal peptide with the Tat apparatus. AB - The DmsD protein is essential for the biogenesis of DMSO reductase in Escherichia coli, and binds the signal peptide of the DmsA subunit, a Tat substrate. This suggests a role as a guidance factor to target pre-DmsA to the translocase. Here, we have analysed the export of fusion proteins in which the DmsA and TorA signal peptides are fused to green fluorescent protein. Both chimeras are efficiently exported to the periplasm in wild-type E. coli cells and we show that their export efficiencies are essentially identical in a mutant lacking DmsD. An authentic Tat substrate, TMAO reductase, is also efficiently exported in the dmsD mutant. The data indicate that DmsD carries out a critical role in DMSO reductase biogenesis/assembly but is not required for the functioning of the DmsA signal peptide. PMID- 12527379 TI - Identification of a MAP65 isoform involved in directional expansion of plant cells. AB - MAP65 comprises a multigene family specific to plants. To see which isoform is utilised for the unique mechanism of cell expansion, uncomplicated by division structures, carrot cells were deprived of auxin whereupon they stopped dividing and elongated instead. During elongation, a MAP65 protein triplet reduced to a single band. Mass spectrometric analysis demonstrated that this corresponded to a single carrot cDNA; it also corresponded to the major protein previously shown to form filamentous cross-bridges between microtubules in vitro. This MAP65 isoform is concluded to have a major role in establishing the parallel microtubule arrays characteristic of cells undergoing directional expansion. PMID- 12527380 TI - Modulation of phosphoinositide 3-kinase activation by cholesterol level suggests a novel positive role for lipid rafts in lysophosphatidic acid signalling. AB - Methyl-beta-cyclodextrin (MbetaCD) was used to explore a role for cholesterol enriched plasma membrane microdomains in coupling lysophosphatidic acid (LPA) stimulation to phosphoinositide 3-kinase (PI3K) activation. Cholesterol depletion strongly inhibited the production of phosphatidylinositol 3,4-bisphosphate and phosphatidylinositol 3,4,5-trisphosphate in Vero cells stimulated with LPA. In agreement, the phosphorylation of Akt/protein kinase B, but not of Erk kinases, was suppressed by MbetaCD. MbetaCD did not interfere with the overall phospholipid metabolism, and its effects were reversed in cholesterol add-back experiments. Finally, PI3K was detected in lipid rafts prepared from control but not MbetaCD-treated cells, suggesting that these microdomains contribute to LPA signalling by compartmentalising component(s) of the PI3K pathway. PMID- 12527381 TI - The transmembranous domain of CD40 determines CD40 partitioning into lipid rafts. AB - Stimulation of CD40 has been previously shown to result in a release of ceramide in small sphingolipid-enriched rafts in the cell membrane [Grassme et al., J. Immunol. 168 (2002) 298-307]. Those rafts fused to larger signaling platforms that served to cluster CD40. Here, we defined molecular mechanisms of CD40 clustering in membrane platforms. To this end, we replaced the transmembranous domain of CD40 with that of CD45, a molecule known to be excluded from lipid rafts. Murine T cells were stably transfected with wild-type CD40 or chimeric CD40/CD45. Flow cytometry confirmed normal binding properties of the mutant to CD40 ligand. Stimulation with CD40 ligand resulted in clustering of wild-type CD40, while the chimeric CD40/45 receptor failed to cluster. This correlated with a deficiency of the chimeric receptor to activate JNK, p38 MAP kinase and SAPK, known signaling molecules of the intracellular pathway initiated by CD40. Forced crosslinking of the CD40/45 chimeric receptor restored, at least partially, these signaling events. The results suggest that the transmembranous domain of CD40 is central for the recruitment to and clustering of CD40 in membrane platforms. PMID- 12527382 TI - Protein kinase C-dependent phosphorylation and mitochondrial translocation of aldose reductase. AB - Although aldose reductase (AR) is a critical participant in osmoregulation, and the metabolism of glucose and aldehydes derived from lipid peroxidation, post translational mechanisms regulating its activity have not been identified. In this paper, we report that stimulation of protein kinase C (PKC) in several cell types induces phosphorylation of AR and translocation of the phosphorylated protein to the mitochondria. In vitro, recombinant AR was directly phosphorylated by activated PKC, suggesting that AR may be an in vivo PKC substrate. Together, these observations reveal a novel link between PKC activation and the regulation of glucose and aldehyde metabolism. PMID- 12527383 TI - Differential incorporation of biotinylated polyamines by transglutaminase 2. AB - Polyamine incorporation or cross-linking of proteins, post-translational modifications mediated by transglutaminase 2 (TGase 2), have been implicated in a variety of physiological functions including cell adhesion, extracellular matrix formation and apoptosis. To better understand the intracellular regulation mechanism of TGase 2, the properties of biotinylated polyamines as substrates for determining in situ TGase activity were analyzed. We synthesized biotinylated spermine (BS), and compared the in vitro and in situ incorporation of BS with that of biotinylated pentylamine (BP), which is an artificial polyamine derivative. When measured in vitro, BP showed a significantly higher incorporation rate than BS. In contrast, in situ incorporation of both BS and BP was not detected even in TGase 2-overexpressed 293 cells. Cells exposed to high calcium showed a marked increase of BP incorporation but not of BS. These data indicate that the in situ activity of TGase 2 gives different results with different substrates, and suggest the possibility of overrepresentation of in situ TGase 2 activity when assayed with BP. Therefore, careful interpretation or evaluation of in situ TGase 2 activity may be required. PMID- 12527384 TI - Diversity of the human erythrocyte membrane sialic acids in relation with blood groups. AB - The composition of the human erythrocyte membrane (RBC) glycoprotein- and glycolipid-bound sialic acids of A, B, AB and O type donors was studied using a new method (Zanetta et al., Glycobiology 11 (2001) 663-676). In addition to Neu5Ac as the major compound, Kdn, Neu5,9Ac(2), Neu5,7Ac(2), Neu (de-N-acetylated Neu5Ac), Neu5Ac8Me, Neu5Ac9Lt, Neu4,5Ac(2), Neu5,8Ac(2)9Lt and Neu5Ac8S were characterised. Among these different compounds, Neu5Ac8Me, Neu5Ac9Lt, Neu4,5Ac(2), Neu5,8Ac(2)9Lt and Neu5Ac8S have never been described and quantitatively determined before in human tissues or cells. Neu5Gc and its O alkylated or O-acylated derivatives were not detected. PMID- 12527385 TI - Control mechanism of JAK/STAT signal transduction pathway. AB - Suppressor of cytokine signaling-1 (SOCS1) was identified as the negative regulator of Janus kinase (JAK) and signal transducer and activator of transcription (STAT) signal transduction pathway. However, the kinetics and control mechanism of the pathway have not yet been fully understood. We have developed the computer simulation of the JAK/STAT pathway. Without nuclear phosphatase, SOCS1's binding to JAK did not cause the decrease in nuclear phosphorylated STAT1. However, without SH2 domain-containing tyrosine phosphatase 2 (SHP-2) or cytoplasmic phosphatase, it did. So nuclear phosphatase is considered to be the most important in this system. By changing parameters of the model, dynamical characteristics and control mechanism were investigated. PMID- 12527386 TI - Tautomeric state of alpha-sarcin histidines. Ndelta tautomers are a common feature in the active site of extracellular microbial ribonucleases. AB - Extracellular fungal RNases, including ribotoxins such as alpha-sarcin, constitute a family of structurally related proteins represented by RNase T1. The tautomeric preferences of the alpha-sarcin imidazole side chains have been determined by nuclear magnetic resonance and electrostatic calculations. Histidine residues at the active site, H50 and H137, adopt the Ndelta tautomer, which is less common in short peptides, as has been found for RNase T1. Comparison with tautomers predicted from crystal structures of other ribonucleases suggests that two active site histidine residues with the Ndelta tautomer are a conserved feature of microbial ribonucleases and that this is related to their ribonucleolytic function. PMID- 12527387 TI - Antibody response to a viral transcriptional regulator. AB - The E2 transcriptional activator of the human papillomavirus regulates the expression of most viral transcripts. Its binding to specific target DNA sequences involves large conformational changes in the interacting macromolecules. The high stability of the E2:DNA complex prompted us to analyze the role of macromolecular interactions and adjuvant emulsions in the appearance of conformation-specific antibodies. We demonstrate that immunization with free or DNA-complexed E2 emulsified in an oil-in-water adjuvant elicits a humoral response shifted to the recognition of discontinuous epitopes. Epitope mapping and functional analysis of the generated anti-E2 mAbs reveals that two separate antibodies populations can be obtained: those able to form a stable ternary complex with protein and DNA, and those which recognize the DNA-binding surface of the transcription factor, interfering with E2 binding to DNA. PMID- 12527388 TI - Kinetic properties of an inulosucrase from Lactobacillus reuteri 121. AB - Inulosucrases catalyze transfer of a fructose moiety from sucrose to a water molecule (hydrolysis) or to an acceptor molecule (transferase), yielding inulin. Bacterial inulin production is rare and a biochemical analysis of inulosucrase enzymes has not been reported. Here we report biochemical characteristics of a purified recombinant inulosucrase enzyme from Lactobacillus reuteri. It displayed Michaelis-Menten type of kinetics with substrate inhibition for the hydrolysis reaction. Kinetics of the transferase reaction is best described by the Hill equation, not reported before for these enzymes. A C-terminal deletion of 100 amino acids did not appear to affect enzyme activity or product formation. This truncated form of the enzyme was used for biochemical characterization. PMID- 12527389 TI - Sex-related differences in neuronal cell survival and signaling in rats. AB - While substantial and compelling evidence exists for sex-related differences in gross and microscopic brain structure, our understanding of the cellular mechanisms that give rise to these sexual dimorphisms is in its infancy. To investigate possible underlying mechanisms for sex-related differences in neuronal development, we examined neuronal survival and transductional signaling in male and female rat primary cortical neuronal cultures. We found that following 14 days in culture, the total number of surviving neurons was significantly higher in female cultures derived from either cortical plate (cp) or ventricular zone (vz), the regions where differentiating (cp) and proliferating (vz) cells are located. In addition, sex-related differences in the levels of phospho-ERK1 and Akt were also observed. Female cortical cultures had significantly higher levels of ERK1 in both cp and vz and higher levels of Akt in cp. No sex-related differences in Bcl-2 were observed. These data suggest that dimorphisms in cell survival may underlie enhanced neuronal survival (or decreased apoptosis) in female brain. Further, the appearance of sex-related differences at cellular and signaling levels in cortical neuronal cultures demonstrates that the effects of gender are not limited to parts of the brain mediating reproduction. PMID- 12527390 TI - Functional organization of the primary motor cortex characterized by event related fMRI during movement preparation and execution. AB - Neuronal recording and neuroimaging studies have shown that the primary motor area (M1) not only participates in motor execution, but is also engaged during movement preparation. The purpose of the present study was to map the distribution of the preparation- and execution-related activity within the contralateral M1 using functional magnetic resonance imaging. Eleven subjects performed a delayed sequential finger movement task, in which a CUE signal indicated a movement sequence in advance of an imperative GO signal. The hemodynamic response related to the CUE and GO signals decreased in a linear fashion across the central sulcus, with activity greater along the lateral extent compared to the medial extent. This decrease was especially evident in the epoch following the CUE. Our data reveal a pattern of functional organization within M1 related to the preparation and execution of movement sequences. PMID- 12527391 TI - Expression of Fc receptor for immunoglobulin M in oligodendrocytes and myelin of mouse central nervous system. AB - Recent advances in neuro-immunology are beginning to elucidate several essential roles of the humoral immunity in both repair and pathogenesis of central nervous system diseases. Immunoglobulin M (IgM) was reported to accelerate the rate of remyelination in a demyelinating disease model, while intrathecal IgM synthesis was shown to predict a more worse disease course in a human demyelinating disease, multiple sclerosis. Molecular mechanisms for either of these IgM functions remain to be studied. Here we report that a recently-found Fc receptor for IgM, namely Fcalpha/muR, is expressed in oligodendrocytes, their precursor cells, and myelin. These expressions were confirmed by immunocytochemistry of cultured cells, Western blot analysis of myelin fractions, and also by immunohistochemistry of mouse forebrains at different ages. Our findings suggest a possible direct interaction between IgM and Fcalpha/muR expressed in oligodendrocytes and myelin. PMID- 12527392 TI - Pyruvic acid cytoprotection against 1-methyl-4-phenylpyridinium, 6 hydroxydopamine and hydrogen peroxide toxicities in vitro. AB - The neuropathology of Parkinson's disease (PD) involves a reduction of endogenous antioxidant enzyme systems, heightened oxidative stress and mitochondrial aberrations in the region of the substantia nigra. Similarly, neurotoxins commonly used to investigate PD pathology include 6-hydroxydopamine (6-OHDA), a powerful hydrogen peroxide (H(2)0(2)) pro-oxidant and 1-methyl-4-phenylpyridinium ion (MPP+), a mitochondrial complex I inhibitor that exerts detrimental effects on cellular energy production. Pyruvic acid is a neuronal metabolic energy fuel that can also rapidly undergo decarboxylation to diffuse H(2)0(2) into H(2)0. In this study, we investigated the effect of pyruvic acid against 6-OHDA, MPP+ and H(2)0(2) toxicity in murine brain neuroblastoma cells. The results obtained indicated that the toxicity of 6-OHDA was inversely related to the autoxidative formation of H(2)0(2). Pyruvic acid exhibited powerful non-enzymatic stoichiometric H(2)0(2) trapping properties, and protected against both 6-OHDA and H(2)0(2) toxicity. While both sodium pyruvate and pyruvate were highly protective against oxidative stress, pyruvate in its free acid form only was protective against MPP+, indicating a requirement for effective transport in order to fuel glycolysis. The protective properties of glucose were compared to pyruvic acid, and the data indicated that glucose did not exhibit antioxidant properties and was effective in blocking MPP+, but not 6-OHDA or H(2)0(2) toxicity. On the other hand, pyruvic acid was protective against all three toxins, and unlike glucose, completely blocked MPP+ toxicity in a combination insult model with up to 500 microM of H(2)0(2). Moreover, the data obtained indicate that pyruvic acid exerts powerful neuroprotective properties by providing simultaneous resistance to oxidative stress and mitochondrial insult. These protective effects are the result of a unique dual property of pyruvic acid with concurrent ability to serve as an effective neuronal energy substrate for glycolysis and to act as an exceptionally powerful antioxidant. PMID- 12527393 TI - Changes in the gene expression of six subtypes of P2X receptors in rat dorsal root ganglion after spinal nerve ligation. AB - Increased purinergic sensitivity of injured sensory neurons suggests the possible involvement of purinoceptors for the generation of pain after nerve injury. To identify the purinoceptors that are involved, the changes in mRNA levels of 6 subtype purinoceptors were examined in the dorsal root ganglia (DRG) of the normal rat and after spinal nerve ligation, using RNase protection assay (RPA). In addition, the P2X(2) containing neurons were examined in the L5 DRG, using an immunohistochemical method. The relative amounts of mRNAs for the six purinoceptor subtypes were in the order of P2X(3)>>P2X(4)>P2X(6)>P2X(5) approximately P2X(2)>P2X(1) in the normal lumbar DRG. After nerve injury, the mRNA of P2X(5) was increased, those of P2X(3) and P2X(6) were decreased, and those of P2X(2) and P2X(4) were unchanged. Immunohistochemical studies, however, showed 23% of the total DRG neurons are P2X(2) positive in the normal L5 DRG, but that increased to 73% after nerve ligation. These data suggest that not only transcriptional but also posttranscriptional changes of multiple purinoceptors might be involved in the enhancement of purinergic sensitivity in injured sensory neurons. PMID- 12527394 TI - In vitro expression of major histocompatibility class I and class II antigens by conditionally immortalized murine neural stem cells. AB - The expression of major histocompatibility complex (MHC) antigens on the surface of cells is intimately linked to in vivo graft survival. It has been previously shown that the conditionally immortalized temperature-sensitive Maudsley hippocampal clone 36 (MHP36) neural stem cells show good long-term graft survival and do not elicit an acute immunological response following transplantation. Here we report that MHP36 cells express both MHC class I and class II antigens when grown in culture under proliferative conditions (33 degrees C), whereas cells with a differentiated morphology in the non-proliferative (37-39 degrees C) condition express low to undetectable levels of either MHC molecules. However, morphologically undifferentiated cells persisting under non-proliferating conditions continued to express both MHC antigens. The downregulation of MHC antigens upon differentiation following cell transplantation could therefore contribute to the graft survival of MHP36 cells. PMID- 12527395 TI - Two distinct inhibitory actions of steroids on cholinoceptor-mediated secretion of catecholamine from guinea-pig adrenal medullary cells. AB - We investigated the effects of dexamethasone, cortisol and aldosterone on responses to nicotine and muscarine in guinea-pig isolated adrenal medullary cells. Nicotine-induced inward currents were reversibly inhibited by these steroids in a dose-dependent and non-competitive manner. These steroids inhibited an increase in [Ca2+](i) in response to nicotine but not muscarine. Muscarine induced catecholamine secretion was inhibited by cortisol and aldosterone but not dexamethasone. Phorbol 12-myristate 13-acetate, a protein kinase C activator, caused catecholamine secretion which was inhibited by cortisol and aldosterone but not dexamethasone. These results suggest that catecholamine secretion induced by cholinoceptor stimulation is inhibited by steroids via two distinct mechanisms; one is the inhibition of nicotinic receptors, another is the inhibition of protein kinase C activation in guinea-pig adrenal medullary cells. PMID- 12527396 TI - Pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid can antagonize the purinoceptor-mediated inhibition of M-current in bullfrog sympathetic neurons. AB - Whole-cell recordings of an M-type potassium current (I(M)) were made from dissociated bullfrog sympathetic neurons. Purinoceptor agonists inhibited I(M) with UTP>ADP>adenosine triphosphate=UDP>ATPgammaS=guanosine triphosphate (GTP)>>amyloid precursor protein (APP)(NH)P as the rank order of potency. The IC(50) was 35 nM for UTP, and 2.6 microM for GTP. Under conditions in which I(M) was abolished by UTP (1 microM), a sulfonic acid derivative, pyridoxalphosphate-6 azophenyl-2',4'-disulfonic acid (PPADS) (30-300 microM) recruited I(M) to 15 to 90% of its control in a reversible and concentration-dependent manner. These results indicate that PPADS can be useful as an antagonist for the purinoceptors presumably P2Y subtypes in amphibian autonomic neurons. PMID- 12527397 TI - Pituitary adenylate cyclase-activating polypeptide modifies the electrical activity of CA1 hippocampal neurons in the rat. AB - The effects of pituitary adenylate cyclase-activating polypeptide (PACAP) on neuronal excitability in the CA1 region of rat hippocampus were studied using in vivo and in vitro electrophysiological techniques. Extracellularly recorded spontaneous firing of CA1 neurons was transiently (2-7 min) increased by PACAP (106+/-32% enhancement, mean+/-SEM, n=11). Using whole-cell patch clamp, PACAP was tested on the resting membrane current of CA1 pyramidal neurons: PACAP activated a slow-onset (20-30 s) and long-lasting (over 20 min) inward current with a mean amplitude of 99+/-34 pA (mean+/-SD, n=8). These results indicate that PACAP induces depolarizing effects on CA1 hippocampal neurons. PACAP-induced long lasting facilitation in the CA1 region might modify neuronal excitability and/or modulate the effect of other neurotransmitters. PMID- 12527398 TI - Non-linear asymmetric interdependencies in the electroencephalogram of healthy term neonates during sleep. AB - The multichannel electroencephalograph (EEG) of six healthy term neonates was recorded during awake as well as during active and quiet sleep. The existence and nature of the interdependencies among the different brain areas were studied by means of a multivariate variant of the surrogate data method. Such interdependencies were then quantified by using the coherence function and a newly developed non-linear index. The results showed that during quiet sleep these interdependencies were mostly non-linear, asymmetric and greater than those found during both awake and active sleep. We conclude that the index might be useful to define patterns of EEG interdependencies in healthy neonates, thereby allowing the early detection of brain dysfunctions. PMID- 12527399 TI - CCAAT/enhancer binding proteins are expressed in the gerbil hippocampus after transient forebrain ischemia. AB - We analyzed CCAAT/enhancer binding protein (C/EBP) family protein levels during reperfusion after a single episode of sublethal forebrain ischemia in the gerbil hippocampus to investigate their expression after ischemia and correlation with neuronal cell death. The common carotid arteries were surgically exposed bilaterally and occluded for 10 min to induce forebrain ischemia in adult Mongolian gerbils. C/EBPalpha, beta, delta, epsilon, zeta protein immunoreactivity was expressed in the hippocampal layer of the CA1 region at 72 h after ischemia and peaked at 96 h. These results appear to correlate with neuronal degeneration as shown by hematoxylin and eosin staining and DNA fragmentation in the terminal transferase biotinylated-UTP nick end labeled method. The present results demonstrate that C/EBP family proteins appear in the selectively vulnerable CA1 pyramidal cell layer in gerbils during neuronal degeneration, and may serve as a signal that neurons are progressing to neuronal cell death and DNA fragmentation. PMID- 12527400 TI - No association between polymorphisms in the neprilysin promoter region and Swedish Alzheimer's disease patients. AB - Amyloid beta-peptide (Abeta) deposition in brain is important in the development of Alzheimer's disease (AD). Neprilysin (NEP) appears to be the major Abeta degrading enzyme in vivo and reduced mRNA levels of NEP correlates with increased plaque density. We hypothesized that alterations in the NEP promoter region may alter NEP expression and thus be involved in the AD process. We investigated three putative important regions in the NEP promoter region; two dinucleotide repeats (CA and GT) and a 480 base pair fragment. With fragment analysis and sequencing, 164 early-onset and 152 late-onset Swedish AD cases and 109 non demented controls were investigated. No significant difference in the distribution of promoter polymorphisms between AD cases and controls were found in this study. PMID- 12527401 TI - Expression of occ1 mRNA in the visual cortex during postnatal development in macaques. AB - We previously reported that the occ1 gene is specifically expressed in the primary visual cortex of adult monkeys in an activity-dependent manner (Tochitani et al., Eur. J. Neurosci., 3, 297-307, 2001). In this report, we compared occ1 mRNA expression in the primary visual cortex during the development of newborn, 3 month-old and adult monkeys. occ1 mRNA was already expressed preferentially in the primary visual cortex of newborn monkeys, but the laminar pattern of occ1 expression in the visual cortex changed as development proceeded. This suggests the possible importance of experience-dependent developmental regulations of occ1 in the developing primary visual cortex. PMID- 12527402 TI - Genetic background of cholesterol gallstone disease. AB - Cholesterol gallstone formation is a multifactorial process involving a multitude of metabolic pathways. The primary pathogenic factor is hypersecretion of free cholesterol into bile. For people living in the Western Hemisphere, this is almost a normal condition, certainly in the elderly, which explains the very high incidence of gallstone disease. It is probably because the multifactorial background genes responsible for the high incidence have not yet been identified, despite the fact that genetic factors clearly play a role. Analysis of the many pathways involved in biliary cholesterol secretion reveals many potential candidates and considering the progress in unraveling the regulatory mechanisms of the responsible genes, identification of the primary gallstone genes will be successful in the near future. PMID- 12527403 TI - Hyposmotic shock: effects on rubidium/potassium efflux in normal and ischemic rat hearts, assessed by 87Rb and 31P NMR. AB - The study evaluated effects of hyposmotic shock on the rate of Rb(+)/K(+) efflux, intracellular pH and energetics in Langendorff-perfused rat hearts with the help of 87Rb- and 31P-NMR. Two models of hyposmotic shock were compared: (1) normosmotic hearts perfused with low [NaCl] (70 mM) buffer, (2) hyperosmotic hearts equilibrated with additional methyl alpha-D-glucopyranoside (Me-GPD, 90 or 33 mM) or urea (90 mM) perfused with normosmotic buffer. Four minutes after hyposmotic shock, Rb(+) efflux rate constant transiently increased approximately two-fold, while pH transiently decreased by 0.08 and 0.06 units, in the first and the second models, respectively, without significant changes in phosphocreatine and ATP. Hyposmotic shock (second model) did not change the rate of Rb(+)/K(+) uptake, indicating that the activity of Na(+)/K(+) ATPase was not affected. Dimethylamiloride (DMA) (10 microM) abolished activation of the Rb(+)/K(+) efflux in the second model; however, Na(+)/H(+) exchanger was not involved, because intracellular acidosis induced by the hyposmotic shock was not enhanced by DMA treatment. After 12 or 20 min of global ischemia, the rate of Rb(+)/K(+) efflux increased by 120%. Inhibitor of the ATP-sensitive potassium channels, glibenclamide (5 microM), partially (40%) decreased the rate constant; however, reperfusion with hyperosmolar buffer (90 mM Me-GPD) did not. We concluded that the shock-induced stimulation of Rb(+)/K(+) efflux occurred, at least partially, through the DMA-sensitive cation/H(+) exchanger and swelling-induced mechanisms did not considerably contribute to the ischemia-reperfusion-induced activation of Rb(+)/K(+) efflux. PMID- 12527404 TI - Inhibition of free radical-induced peroxidation of rat liver microsomes by resveratrol and its analogues. AB - Resveratrol (3,5,4'-trans-trihydroxystilbene) is a natural phytoalexin present in grapes and red wine, which possesses a variety of biological activities including antioxidative activity. To find more efficient antioxidants by structural modification, resveratrol analogues, that is, 3,4-dihydroxy-trans-stilbene (3,4 DHS), 4,4'-dihydroxy-trans-stilbene (4,4'-DHS), 4-hydroxy-trans-stilbene (4-HS) and 3,5-dihydroxy-trans-stilbene (3,5-DHS), were synthesized and their antioxidant activity studied for the free radical-induced peroxidation of rat liver microsomes in vitro. The peroxidation was initiated by either a water soluble azo compound 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH) or Fe(2+)/ascorbate, and monitored by oxygen uptake and formation of thiobarbituric acid reactive substances (TBARS). It was found that all of these trans-stilbene derivatives are effective antioxidants against both AAPH- and iron-induced peroxidation of rat liver microsomes with an activity sequence of 3,4-DHS>4,4' DHS>resveratrol>4-HS>3,5-DHS. The remarkably higher antioxidant activity of 3,4 DHS is discussed. PMID- 12527405 TI - Reduction in cardiac mitochondrial calcium loading capacity is observable during alpha-naphthylisothiocyanate-induced acute cholestasis: a clue for hepatic derived cardiomyopathies? AB - Cardiovascular changes of still obscure origin are sometimes correlated with co existing liver diseases, as cholestasis. The aim of this work was to examine and compare cardiac mitochondrial bioenergetics and calcium loading capacity from rats injected with a single dose of alpha-naphthylisothiocyanate (ANIT), a cholestasis-inducing compound. Forty-eight hours after ANIT administration, blood samples were collected and markers for hepatic disease were determined. Heart mitochondria from both control and ANIT-injected rats were isolated and subjected to biochemical characterization, including the susceptibility to the calcium dependent permeability transition. The results showed that cardiac mitochondria from cholestatic animals did not have significant changes in respiratory parameters or in the basal levels of adenine nucleotide. The most impressive result from this work was that cardiac mitochondria from ANIT-injected animals had a lower calcium loading capacity. The prevention of this property by cyclosporin-A, a specific inhibitor of the mitochondrial permeability transition, showed that this phenomenon was reason for the reduced calcium loading capacity in ANIT-injected animals. The results suggest that, during the development of ANIT-induced cholestasis, heart mitochondria loose their default ability to buffer calcium. Our results may contribute to explain the occurrence of cardiomyopathies sometimes associated with cholestatic disease. PMID- 12527406 TI - Characterization of hydrogen peroxide removal reaction by hemoglobin in the presence of reduced pyridine nucleotides. AB - Hydrogen peroxide removal rates by hemoglobin were enhanced in the presence of reduced pyridine nucleotides. The species which had the activity to oxidize pyridine nucleotides was purified from human blood and identified as hemoglobin A. Hydrogen peroxide removal rates by hemoglobin A without reduced pyridine nucleotides at 0.2 mM hydrogen peroxide were 0.87+/-0.11 micromol/s/g hemoglobin, and the removal rates using 0.2 mM NADH and NADPH were 2.02+/-0.20 and 1.96+/ 0.31 micromol/s/g hemoglobin, respectively. We deduced that the removal reaction by hemoglobin included formations of methemoglobin and the ferryl radical and reduction of the latter with pyridine nucleotides. The hydrogen peroxide removal ability by hemoglobin was less than that by catalase but was larger than that by glutathione peroxidase-glutathione reductase system at 0.2 mM hydrogen peroxide. Under acatalasemic conditions, it was suggested that NAD(P)H were important factors to prevent the oxidative degradation of hemoglobin. PMID- 12527407 TI - Antibody production in early life supported by maternal lymphocyte factors. AB - To examine the influence of maternal lymphocyte factors on the immune responses in offspring in early life, antibody production in neonates born to either normal or lymphocyte-deficient mothers was analyzed. Recombination activating gene (Rag) 2(+/-) mouse neonates born to Rag-2(+/+), Rag-2(+/-)or Rag-2(-/-)mothers were injected with goat anti-mouse IgD antiserum, and IgE and IgG(1) production was evaluated. The levels of IgE and IgG(1) were higher in the pups born to Rag 2(+/+)and Rag-2(+/-) dams than to lymphocyte-deficient Rag-2(-/-) dams. The enhanced antibody production in the former compared with the latter neonates was also found following immunization with ovalbumin or TNP-Ficoll. Thus, the presence of maternal lymphocyte factors was suggested in neonates that augmented antigen-specific antibody production in both T cell-dependent and -independent pathways. A reduction in antibody production was observed in normal neonates when they were foster-nursed by Rag-2(-/-) mothers. Thus, the maternal lymphocyte factors enhancing the immune responses in newborns were shown to be present in breast-milk. PMID- 12527409 TI - Spermine induces cataract and 43-kDa protein that binds spermine possibly participates in the cataract formation. AB - Among polyamines (putrescine, spermidine, and spermine), spermine specifically induces cataract in an organ cultured lens. Spermine uptake nearly paralleled the cataract formation. When polyamines were added to lens soluble proteins, spermine specifically induced turbidity. When lens soluble proteins were separated by gel chromatography, heavy-molecular-weight protein (HMW, high molecular form of alpha crystallin) and proteins between betaH- and betaL-crystallin fractions reacted with spermine and aggregated. SDS-polyacrylamide gel electrophoresis of the aggregated proteins showed that 43-kDa lens protein was commonly observed in both aggregates. Spermine-affinity chromatography of the total soluble proteins showed the binding of HMW protein to the gel and the chromatogram of the second turbidity peak in the gel chromatography showed the binding of 43-kDa protein. These results indicated that 43-kDa protein, which is present as a subunit in HMW and also in free form, binds spermine and induces turbidity of lens soluble proteins and produces cataract in a cultured lens. PMID- 12527408 TI - Inhibition of protein kinase C by resveratrol. AB - Evidence is emerging that resveratrol (RV), a polyphenolic phytoaxelin present in dietary sources including red wine, may protect against atherosclerosis and cardiovascular disease by enhancing the integrity of the endothelium. In this study, the possibility that such beneficial effects of RV may arise from a modulation of protein kinase C (PKC)-mediated signaling was investigated by determining the effects of RV on the in vitro activities of PKC isozymes. It was found that the Ca(2+)-dependent activities of membrane-associated PKCalpha induced by either phorbol ester or diacylglycerol were potently inhibited by RV, each with an IC(50) of approximately 2 microM. The inhibitory effect of RV was also observed for conventional PKCbetaI, whereas the activities of novel PKC epsilon and atypical PKCzeta were each unaffected. The inhibition of PKCalpha activity was found to be competitive with respect to phorbol ester concentration but noncompetitive with respect to Ca(2+) and phosphatidylserine concentrations, suggesting that the RV may compete for phorbol ester-binding to the C1 domains. Supporting this, it was found that RV bound to a fusion peptide containing the C1A and C1B domains of PKCalpha. Similar to the effects of diacylglycerol and phorbol ester, the interaction of RV with the C1 domains induced the association of PKCalpha with membrane lipid vesicles, although this did not result in activation. Overall, the results suggest that the inhibitory effect of RV on PKC activity, and therefore on the associated signaling networks, may, in part, underlie the mechanism(s) by which this agent exerts its beneficial effects on endothelial and cardiovascular function. Furthermore, the effects of RV on these signaling networks are predicted to differ according to the cellular localization and the regulating PKC isozyme. PMID- 12527410 TI - High in comparison with low tidal volume ventilation aggravates oxidative stress induced lung injury. AB - Ventilator settings influence the development and outcome of acute lung injury. This study investigates the influence of low versus high tidal volume (V(t)) on oxidative stress-induced lung injury. Isolated rabbit lungs were subjected to one of three ventilation patterns (V(t)-positive end-expiratory pressure, PEEP): LVZP (6 ml/kg-0 cm H(2)O), HVZP (12 ml/kg-0 cm H(2)O), LV5P (6 ml/kg-5 cm H(2)O). These ventilation patterns allowed a comparison between low and high V(t) without dependence on peak inspiratory pressure (PIP). Infusion of hypochlorite (1000 nmol/min) or buffer (control) was started at t=0 min. Pulmonary artery pressure (PAP), PIP and weight were continuously recorded. Capillary filtration coefficient [K(f,c) (10(-4) ml s(-1) cm H(2)O(-1) g(-1))] was gravimetrically determined (-15/30/60/90/120 min).PIP averaged 5.8+/-0.6/13.9+/-0.6/13.9+/-0.4 cm H(2)O in the LVZP, HVZP and LV5P groups. PIP, K(f,c) or PAP did not change in control groups, indicating that none of the ventilation patterns caused lung injury by themselves. Hypochlorite-induced increase in K(f,c) but not hypochlorite-induced increase in PAP, was significantly attenuated in the LVZP /LV5P- versus the HVZP-group (K(f,c,max.) 1.0+/-0.23/1.4+/-0.40 versus 3.2+/ 1.0*). Experiments with hypochlorite were terminated due to excessive edema (>50 g) at 97+/-2.2/94.5+/-4.5 min in the LVZP-/LV5P-group versus 82+/-3.8* min in the HVZP-group (*: P<0.05). Low V(t) attenuated oxidative stress-induced increase in vascular permeability independently from PIP and PEEP. PMID- 12527411 TI - Augmented lipopolysaccharide-induction of the histamine-forming enzyme in streptozotocin-induced diabetic mice. AB - Disorders of the microcirculation and reduced resistance to infection are major complications in diabetes. Histamine enhances capillary permeability, and may also reduce cellular immunity. Here we demonstrate that streptozotocin (STZ) induced diabetes in mice not only enhances the activity of the histamine-forming enzyme, histidine decarboxylase (HDC), but also augments the lipopolysaccharide (LPS)-induced elevation of HDC activity in various tissues, resulting in a production of histamine. The augmentation of HDC activity occurred as early as 2 days after STZ injection, but was not seen in nondiabetic mice. When given to STZ treated mice, nicotinamide, an inhibitor of poly(ADP-ribose) synthetase, reduced both the elevation of blood glucose and the elevations of HDC activity and histamine production. These results suggest that hyperglycemia may initiate a sequence of events leading not only to an enhancement of basal HDC activity, but also to a sensitization of mice to the HDC-inducing action of LPS. We hypothesize that bacterial infections and diabetic complications may mutually exacerbate one another because both involved an induction of HDC. PMID- 12527412 TI - Effect and possible role of Zn treatment in LEC rats, an animal model of Wilson's disease. AB - The effect of oral zinc (Zn) treatment was studied in the liver, kidneys and intestine of Long-Evans Cinnamon (LEC) rats in relation to metals interaction and concentration of metallothionein (MT) and glutathione (GSH). We also investigated the change in the activity of antioxidant enzymes and determined the biochemical profile in the blood and metal levels in urine. We showed that the Zn-treated group had higher levels of MT in the hepatic and intestinal cells compared to both untreated and basal groups. Tissue Zn concentrations were significantly higher in the Zn-treated group compared to those untreated and basal, whereas Cu and Fe concentrations decreased. The antioxidant enzyme activities in the Zn treated group did not change significantly with respect to those in the basal group, except for hepatic glutathione peroxidase activity. Moreover, the biochemical data in the blood of Zn-treated group clearly ascertain no liver damage. These observations suggest an important role for Zn in relation not only to its ability to compete with other metals at the level of absorption in the gastrointestinal tract producing a decrease in the hepatic and renal Cu and Fe deposits, but also to MT induction as free radical scavenger. PMID- 12527413 TI - Glyceraldehyde-3-phosphate dehydrogenase activity as an independent modifier of methylglyoxal levels in diabetes. AB - Methylglyoxal (MG) may be an important cause of diabetic complications. Its primary source is dihydroxyacetone phosphate (DHAP) whose levels are partially controlled by glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Using a human red blood cell (RBC) culture, we examined the effect of modifying GAPDH activity on MG production. With the inhibitor koningic acid (KA), we showed a linear, concentration-dependent GAPDH inhibition, with 5 microM KA leading to a 79% reduction of GAPDH activity and a sixfold increase in MG. Changes in redox state produced by elevated pH also resulted in a 2.4-fold increase in MG production at pH 7.5 and a 13.4-fold increase at pH 7.8. We found substantial inter-individual variation in DHAP and MG levels and an inverse relationship between GAPDH activity and MG production (R=0.57, P=0.005) in type 2 diabetes. A similar relationship between GAPDH activity and MG was observed in vivo in type 1 diabetes (R=0.29, P=0.0018). Widely varying rates of progression of diabetic complications are seen among individuals. We postulate that modification of GAPDH by environmental factors or genetic dysregulation and the resultant differences in MG production could at least partially account for this observation. PMID- 12527414 TI - Inhibition of succinic semialdehyde dehydrogenase activity by alkenal products of lipid peroxidation. AB - Lipid peroxidation causes the generation of the neurotoxic aldehydes acrolein and 4-hydroxy-trans-2-nonenal (HNE). These products are elevated in neurodegenerative diseases and acute CNS trauma. Previous studies demonstrate that mitochondrial class 2 aldehyde dehydrogenase (ALDH2) is susceptible to inactivation by these alkenals. In the liver and brain another mitochondrial aldehyde dehydrogenase, succinic semialdehyde dehydrogenase (SSADH/ALDH5A1), is present. In this study, we tested the hypothesis that aldehyde products of lipid peroxidation inhibit SSADH activity using the endogenous substrate, succinic semialdehyde (SSA, 50 microM). Acrolein potently inhibited SSADH activity (IC(50)=15 microM) in rat brain mitochondrial preparations. This inhibition was of an irreversible and noncompetitive nature. HNE inhibited activity with an IC(50) of 110 microM. Trans 2-hexenal (HEX) and crotonaldehyde (100 microM each) did not inhibit activity. These data suggest that acrolein and HNE disrupt SSA metabolism and may have subsequent effects on CNS neurochemistry. PMID- 12527415 TI - A single site in human beta-hexosaminidase A binds both 6-sulfate-groups on hexosamines and the sialic acid moiety of GM2 ganglioside. AB - Human beta-hexosaminidase A (Hex A) (alphabeta) is composed of two subunits whose primary structures are approximately 60% identical. Deficiency of either subunit results in severe neurological disease due to the storage of GM2 ganglioside; Tay Sachs disease, alpha deficiency, and Sandhoff disease, beta deficiency. Whereas both subunits contain active sites only the alpha-site can efficiently bind negatively charged 6-sulfated hexosamine substrates and GM2 ganglioside. We have recently identified the alphaArg(424) as playing a critical role in the binding of 6-sulfate-containing substrates, and betaAsp(452) as actively inhibiting their binding. To determine if these same residues affect the binding of the sialic acid moiety of GM2 ganglioside, an alphaArg(424)Gln form of Hex A was expressed and its kinetics analyzed using the GM2 activator protein:[3H]-GM2 ganglioside complex as a substrate. The mutant showed a approximately 3-fold increase in its K(m) for the complex. Next a form of Hex B (betabeta) containing a double mutation, betaAspLeu(453)AsnArg (duplicating the alpha-aligning sequences), was expressed. As compared to the wild type (WT), the mutant exhibited a >30-fold increase in its ability to hydrolyze a 6-sulfated substrate and was now able to hydrolyze GM2 ganglioside when the GM2 activator protein was replaced by sodium taurocholate. Thus, this alpha-site is critical for binding both types of negatively charge substrates. PMID- 12527416 TI - Inhibition of the alpha-ketoglutarate dehydrogenase complex alters mitochondrial function and cellular calcium regulation. AB - Mitochondrial dysfunction occurs in many neurodegenerative diseases. The alpha ketoglutarate dehydrogenase complex (KGDHC) catalyzes a key and arguably rate limiting step of the tricarboxylic acid cycle (TCA). A reduction in the activity of the KGDHC occurs in brains and cells of patients with many of these disorders and may underlie the abnormal mitochondrial function. Abnormalities in calcium homeostasis also occur in fibroblasts from Alzheimer's disease (AD) patients and in cells bearing mutations that lead to AD. Thus, the present studies test whether the reduction of KGDHC activity can lead to the alterations in mitochondrial function and calcium homeostasis. alpha-Keto-beta-methyl-n-valeric acid (KMV) inhibits KGDHC activity in living N2a cells in a dose- and time dependent manner. Surprisingly, concentration of KMV that inhibit in situ KGDHC by 80% does not alter the mitochondrial membrane potential (MMP). However, similar concentrations of KMV induce the release of cytochrome c from mitochondria into the cytosol, reduce basal [Ca(2+)](i) by 23% (P<0.005), and diminish the bradykinin (BK)-induced calcium release from the endoplasmic reticulum (ER) by 46% (P<0.005). This result suggests that diminished KGDHC activities do not lead to the Ca(2+) abnormalities in fibroblasts from AD patients or cells bearing PS-1 mutations. The increased release of cytochrome c with diminished KGDHC activities will be expected to activate other pathways including cell death cascades. Reductions in this key mitochondrial enzyme will likely make the cells more vulnerable to metabolic insults that promote cell death. PMID- 12527417 TI - Mitochondrially mediated synergistic cell killing by bile acids. AB - The accumulation of endogenous bile acids contributes to hepatocellular damage during cholestatic liver disease. To examine the controversy regarding the therapeutic use of ursodeoxycholate (UDCA) in cholestatic patients, we investigated the possible cytoprotection or synergistic effects of UDCA against chenodeoxycholate (CDCA)-induced injury to isolated rat hepatocytes. Our aim was to investigate the role of the mitochondrial permeability transition (MPT) in the mechanism of cytotoxicity caused by UDCA plus CDCA. Although not toxic by itself, UDCA potentiated the mitochondrial depolarization, ATP depletion and cell killing caused by CDCA. Fructose maintained ATP levels and prevented bile acid-induced cell killing. Cyclosporine A (CyA), a potent inhibitor of the MPT, substantially reduced mitochondrial depolarization, ATP depletion and cell killing caused by CDCA. Our results demonstrate that the synergistic cytotoxicity by UDCA plus CDCA is mediated by impairment of mitochondrial function, an event that is expressed via induction of the MPT. PMID- 12527419 TI - Genomic and molecular characterization of CL-43 and its proximal promoter. AB - Collectins are part of the innate immune system as they bind nonself glycoconjugates on the surface of microorganisms and inhibit infection by direct neutralization, agglutination or opsonization of the invaders. Conglutinin and CL 43 are serum proteins that have only been found and characterized in Bovidae. We have studied molecular and genomic characteristics of CL-43 to identify polymorphisms that might be associated with disease-susceptible phenotypes or other traits in cattle, and to elucidate how the Bovidae may benefit from possessing additional collectins. Screening a bovine cDNA library resulted in the isolation of two plasmid clones that encoded the entire translated sequence of CL 43. The 5'-untranslated end and start point of transcription were identified by 5'-RACE and showed that the mRNA transcript comprises either 1326 or 1241 nucleotides because of alternative splicing. Both transcripts encode a protein of 321 amino acids including a signal peptide of 20 residues. Characterization of two overlapping genomic lambda phage clones showed that the gene comprised seven exons spanning 8.5 kbp. The CL-43 gene, like the conglutinin gene, was mapped to Bos taurus chromosome 28 at q1.8. The CL-43 promoter has 96% identity with the conglutinin promoter recently described by us, and the assignment of potential cis-regulatory elements shows that several hepatic transcription factors may regulate transcription in the acute phase response and in response to metabolic changes. PMID- 12527420 TI - Isolation and characterisation of tilapia beta-actin promoter and comparison of its activity with carp beta-actin promoter. AB - The regulatory sequence including proximal promoter, untranslated exon 1 and intron 1 of the beta-actin gene from tilapia (Oreochromis niloticus) has been isolated and spliced to a beta-galactosidase reporter gene to test its activity. Comparisons of promoter activity have been carried out with three different constructs: (1) 1.6 kb tilapia beta-actin regulatory sequence, (2) 1.5 kb carp beta-actin regulatory sequence, and (3) 4.7 kb carp beta-actin regulatory sequence. Although the 1.6 kb tilapia beta-actin regulatory sequence gave slightly different expression patterns in tilapia embryos assayed by in situ X gal staining, no difference was observed in expression level when the tilapia sequence was compared with the 4.7 kb carp beta-actin regulatory sequence by quantitative assay. In comparison with the 1.5 kb carp beta-actin regulatory sequence, the 1.6 kb tilapia beta-actin regulatory sequence gave higher expression levels in tilapia embryos, while a reverse result was observed in zebrafish embryos. In cell transfection experiments, the 1.6 kb tilapia beta actin regulatory sequence showed three to four times better activity in blue gill cells than either the 4.7 kb carp beta-actin or the 1.5 kb carp beta-actin regulatory sequences. The 1.6 kb tilapia beta-actin regulatory sequence also drove higher reporter gene activity in somatic cells of tilapia than did the 4.7 kb carp beta-actin regulatory sequence following direct injection of constructs into muscle. Therefore, taken together, the data demonstrate that the tilapia beta-actin promoter can be used as an efficient regulatory sequence to produce autotransgenic tilapia. PMID- 12527421 TI - Sequence of the gene for Clostridium botulinum type B neurotoxin associated with infant botulism, expression of the C-terminal half of heavy chain and its binding activity. AB - Previously, we demonstrated that the neurotoxin of strain 111 (111/NT) associated with type B infant botulism showed antigenic and biological properties different from that (Okra/NT) produced by a foodborne botulism-related strain, Okra. In this study, the neurotoxin genes of 111/NT and Okra/NT were amplified and the sequences determined. The nucleotide sequences of the genes for both neurotoxins possessed an open reading frame of 3873 bp that encoded 1291 amino acids. The identities of nucleotide sequences and amino acid sequences were 97.6% and 95.7%, respectively. The ratio of nonsynonymous to synonymous substitutions was 0.47. The amino acid substitutions between 111/NT and Okra/NT occurred mainly in the domain of the C-terminal half of heavy chain (H(C)) responsible for binding to its receptor complex of protein and ganglioside. To characterize the binding capability of the H(C), recombinant genes for the H(C) and two hybrid H(C) in which one half of Okra/NT was replaced by the homologous half of 111/NT were constructed and expressed in Escherichia coli. The binding activity of the recombinant H(C) of 111/NT to the protein receptor synaptotagmin II, in the presence of ganglioside GT1b, was 4.2-fold less than Okra/NT, consistent with the corresponding two NTs. The use of hybrid H(C) revealed that mutation of 23 residues in carboxy terminal half of H(C) (1029-1291) of Okra/NT could be attributed to the lower binding activity of 111/NT and thus the differences in binding affinity between the two BoNT/B. PMID- 12527422 TI - Crystallization and preliminary X-ray analysis of anti-cancer agent 3-(9 acridinylamino)-5-(hydroxymethyl)aniline complexed with the DNA hexamer d(CGTACG)2. AB - 3-(9-Acridinylamino)-5-(hydroxymethyl)aniline (AHMA) is an anti-cancer agent with significant efficacy against murine leukemia and solid tumors. As a DNA topoisomerase inhibitor, AHMA is proposed to form a ternary complex with DNA and topoisomerase and bind to DNA in an intercalative manner. In order to understand the interactions between AHMA and DNA and study the structure-function relationship of amsacrine analogue, the AHMA-d(CGTACG)(2) complex was crystallized using the sitting-drop vapor-diffusion method. The native crystals diffract to 2.9-A resolution and belong to space group P3(1)21 or P3(2)21 with unit-cell parameters a=b=57.52, c=122.17 A when analyzed using Cu Kalpha radiation. Patterson map indicates that in the crystal, the directions of the DNA base stacking are nearly perpendicular to the c-axis of the crystal unit cell. PMID- 12527423 TI - Characterization of the 5'-flanking fragment of the human GM3-synthase gene. AB - To investigate the transcriptional regulation of human GM3-synthase, a 5' flanking fragment of 1379 bp was cloned by a PCR-based procedure. Analysis of the human genomic sequence showed that the gene consists of seven exons, locates at chromosome 2, and spans over 62 kb. There are a number of potential consensus binding sites in the cloned promoter region, but TATA and CCAAT boxes were not found in the promoter. Primer extension analysis identified two transcription start sites approximately 11 and 57 bp upstream of the exon 1. The transcription activity of the promoter was assessed in human HeLa cells by transient transfection. Of the fragments assayed, the proximal 409 bp fragment exhibits the highest transcription activity. Transcription factors that bound to the 409 bp fragment were pulled down by DNA-coupled magnetic beads. Identities of the pull down proteins were determined by array analysis. Eight transcription factors were identified, which might either bind to the proximal region or be recruited as co activators of the transcription factor complexes. PMID- 12527424 TI - Differential expression of plasminogen activator inhibitor-1, tumor necrosis factor-alpha, TNF-alpha converting enzyme and ADAMTS family members in murine fat territories. AB - Our objective was to investigate expression of A disintegrin and metalloproteinase (ADAM) and ADAM proteins with a thrombospondin (TS) motif (ADAMTS) family members in adipose tissue of lean and obese mice. Five-week-old male mice were kept on standard chow (SFD) or on high fat diet (HFD) for 15 weeks, and subcutaneous (SC) and gonadal (GON) adipose tissue, as well as mature adipocytes and stromal-vascular (S-V) cells were harvested. mRNA levels of plasminogen activator inhibitor-1 (PAI-1), tumor necrosis factor-alpha (TNF alpha), ADAM-17 (TACE or TNF-alpha converting enzyme), ADAMTS-1 and ADAMTS-8 were quantified in isolated adipose tissues and cell fractions, and during differentiation of murine preadipocytes. The HFD resulted in a significantly enhanced weight of isolated SC and GON fat pads, and in enhanced blood levels of glucose, cholesterol and PAI-1. ADAM-17, TNF-alpha, PAI-1, ADAMTS-1 and ADAMTS-8 mRNA were detected in both SC and GON adipose tissue of lean mice (SFD). In SC adipose tissue of obese mice (HFD), the expression of ADAM-17 and PAI-1 was enhanced and that of ADAMTS-1 reduced, whereas in GON adipose tissue expression of TNF-alpha was enhanced and that of ADAMTS-8 reduced. In lean and obese mice, expression of ADAM-17, ADAMTS-1 and ADAMTS-8 was higher in the S-V cell fraction than in mature adipocytes. During differentiation of murine 3T3-F442A preadipocytes, expression of ADAM-17 and ADAMTS-1 remained virtually unaltered, whereas that of ADAMTS-8 decreased as adipocytes matured. Several ADAM and ADAMTS family members are expressed in adipose tissue and during differentiation of preadipocytes. Modulation of their expression upon development of obesity is adipose tissue-dependent. PMID- 12527425 TI - Sequence analysis, transcriptional and posttranscriptional regulation of the rice vdac family. AB - The voltage-dependent anion-selective channel (VDAC) is a mitochondrial outer membrane ion channel. Different isoforms exist in plants but information about their specific role remains to be established. Our purpose is to find out the structural features common to three rice VDAC isoforms and to investigate their (post)transcriptional regulation in response to an osmotic stress. Two new cDNAs encoding mitochondrial VDAC from rice (Oryza sativa) were isolated, sequenced and characterized: a phylogenetic reconstruction permitted identification of orthologues in Poaceae and computer-based analyses predicted 18 transmembrane beta-strands, one amphipathic alpha-helix and two different phosphorylation motifs. The expression of three rice vdac genes was investigated. Northern blot analyses indicated that they were expressed in all plant tissues. There was a differential expression of osvdac1 and osvdac3, whereas osvdac2 was homogeneously expressed in all tissues. No change in vdac expression was observed under an osmotic stress. However, a fast-enhanced expression of vdac was observed in roots during the recovery period after stress release. This enhanced expression is not correlated to the amount of VDAC protein detected in roots suggesting a posttranscriptional regulation. PMID- 12527426 TI - Cloning and characterization of the rat Hsf2 promoter: a critical role of proximal E-box element and USF protein in Hsf2 regulation in different compartments of the brain. AB - The complex patterns of tissue-, cell type- and developmental stage-specific expression of heat shock factor 2 (Hsf2) raise a question of how this can be achieved for this ubiquitous transcription factor. To explore molecular mechanisms responsible for the regulated expression of Hsf2, a 2638-bp 5' flanking region of the rat Hsf2 gene was cloned and characterized. Since the brain represents one of the most complicated organs composed of several regions with different cell types, differential regulation of Hsf2 in various brain regions was investigated in detail. Results show that the major transcription initiation site of the Hsf2 gene is located at cytosine-155 relative to the translation initiation site. The E-box element located immediate upstream of the transcription initiation site was demonstrated to be critical for Hsf2 promoter activity, and the upstream stimulatory factor (USF) protein was identified as the major E-box binding protein. That the only two base exchange of the E-box core sequences from CACGTG to CACGGT severely impaired Hsf2 promoter activity and completely eliminated USF binding clearly demonstrated that the specific binding of USF to E-box is critical for Hsf2 promoter activity. Here we demonstrated that the Hsf2 expression levels varied significantly in different brain regions. We also demonstrated that Hsf2 expression levels in various brain regions relatively correlated with the E-box binding activity of USF. Based on these results, we suggest that E-box binding activity of USF protein may act as one of the major regulators of Hsf2 expression in situ although a possible involvement of other transcription factors cannot be ruled out. The presence of several transcription factor binding sites of biological importance in the Hsf2 promoter suggests that identifying the interplay of USF and these factors should help further elucidate the molecular mechanisms of tissue-, cell type- and developmental stage-specific expression of Hsf2. PMID- 12527427 TI - Seabream growth hormone receptor: molecular cloning and functional studies of the full-length cDNA, and tissue expression of two alternatively spliced forms. AB - A full-length clone of the growth hormone receptor (GHR) was isolated from a cDNA library constructed from the liver of black seabream (Acanthopagrus schlegeli). The seabream GHR (sbGHR) cDNA sequence encodes a transmembrane protein of 640 amino acids (aa) possessing the characteristic motifs and architectural design of GHRs of other species. When compared to the other fish GHRs, it is most homologous to another marine fish species, the turbot, where the aa identity is 79.3%. But the sbGHR sequence is more remotely related to the goldfish GHR (51.6% aa identity) and the salmonid GHRs (approximately 46-48% aa identities). Phylogenetic comparison with other known GHRs indicates that the fish GHRs constitute a distinct group among the different vertebrate classes. The aa identities between sbGHR and other GHRs are low, being around 40% with mammalian GHRs, around 45% with avian and reptilian GHRs, and less than 35% with Xenopus GHR. CHO cells transfected with the sbGHR cDNA can be stimulated to proliferate by recombinant seabream growth hormone (sbGH). In addition, the transfected cells can transactivate a co-expressed mammalian serine protease inhibitor (Spi) 2.1 promoter upon stimulation by sbGH. These functional assays indicated that the fish receptor can interact with its homologous ligand to evoke the downstream post-receptor events. Reverse transcription-polymerase chain reaction (RT-PCR) and genomic PCR using a pair of gene-specific primers revealed the expression of two alternatively spliced forms of sbGHR in various tissues of the fish. A 93-bp intron, unique to the sbGHR gene and not found in any other known GHR genes, is alternatively spliced to give rise to two forms of receptor mRNA transcripts. The two forms of the receptor are differentially expressed among the different tissues of the fish. PMID- 12527428 TI - Structure of gene flanking regions and functional analysis of sequences upstream of the rat hsp70.1 stress gene. AB - We present structural and comparative analysis of the flanking regions of the rat hsp70.1 stress gene. Several repetitive sequences, microsatellites and short interspersed repetitive elements (SINEs) were found, as well as a significant gap in the 3' UTR, as compared to the orthologous mouse gene. We also show that the complex microsatellite region composed of partially overlapping inverted repeat and long homopurine-homopyrimidine sequence, which is localized 1.8 kbp upstream of the transcription start site, is capable to adopt non-B DNA structures (an H DNA and a cruciform structure) in vitro. Functional analysis performed with the use of various fragments of the 5'end flanking regions ligated to the chloramphenicol acetyltransferase (CAT) reporter gene revealed a crucial role of cooperation between heat shock element (HSE) regulatory sequences, while none of the three HSEs alone is able to drive efficient heat induced transcription of the reporter gene. We also found that the microsatellite region does not influence transcription by itself, however, it abolishes the effect of the adjacent putative silencing element. To our knowledge, this is a first extensive structural and functional analysis of the promoter region of the mammalian heat inducible hsp70i gene localized distally to the hsp70-related spermatid-specific gene in the major histocompatibility complex III. PMID- 12527429 TI - Expression of human TFIIA subunits in Saccharomyces cerevisiae identifies regions with conserved and species-specific functions. AB - The transcription factor TFIIA stabilizes the interaction between the TATA binding protein (TBP) and promoter DNA and facilitates activator function. In yeast, TFIIA is composed of large (TOA1) and small (TOA2) subunits that interact to form a beta-barrel domain and a helix bundle domain. Here we report plasmid shuffle experiments showing that the human subunits (TFIIAalpha/beta, ALF, and TFIIAgamma) are not able to support growth in yeast and that the failure is associated with morphological abnormalities related to cell division. To determine the regions responsible for species specificity, we examined a series of chimeric yeast-human subunits. The results showed that yeast-human hybrids that contained the N-termini of TFIIAgamma or TFIIAalpha/beta were viable, presumably because they could form a functional interspecies alpha-helical bundle. Likewise, a TOA1 hybrid that contained the nonconserved internal region from TFIIAalpha/beta also had no effect on TFIIA function. However, hybrids that contained the acidic region III or C-terminal region IV from TFIIAalpha/beta grew more slowly than the wild-type TOA1 subunit, and if both regions were exchanged, this effect was far more severe. Although these hybrids exchanged sequences which are involved in beta-barrel formation and interactions with TBP, they were all active in a TBP-dependent mobility shift assay. The results suggest that the growth phenotypes of these hybrids might be due to a failure to interact with components of the yeast transcription machinery other than TBP. Finally, we show that sequences from region III of TFIIA large subunits fall into classes that are either highly acidic or that are divergent and nonacidic, and provide the first evidence to suggest that, at least in yeast, this region is important for TFIIA function. PMID- 12527430 TI - Expression analysis and mitochondrial targeting properties of the chicken manganese-containing superoxide dismutase. AB - Manganese-containing superoxide dismutase (MnSOD) is a major detoxifying enzyme that functions in cellular oxygen metabolism by converting O(2)(-) to H(2)O(2). A cDNA encoding the chicken MnSOD (cMnSOD) has been isolated from a chicken embryo fibroblast (CEF) cell cDNA library. The cloned cMnSOD is 1102 bp in length with an open reading frame (ORF) of 224 amino acids that includes a 26-amino-acid 5' proximal mitochondrial targeting sequence (MTS). The mature 198-amino-acid region of the cMnSOD is highly conserved among various mammalian species. Two cMnSOD mRNA species (1.2 and 1.0 kb) were expressed in most of the tissues and organs analyzed, with the highest expression levels found in brain, kidney, and heart tissues. Compared to earlier stages of development, expression of cMnSOD was highest in day 13 embryonic heart tissue, and was maintained until post-hatch. Exogenously introduced cMnSOD-GFP fusion constructs (which included the MTS) clearly accumulated in the mitochondria of chicken cells, as expected. Surprisingly, the cMnSOD MTS signal, which displays little similarity to mammalian MTS sequences, enabled cMnSOD-GFP fusion proteins to target mitochondria not only from different cell types (fibroblastic and epithelial), but from a number of mammalian species (human, mouse, and pig). This suggests that specific amino acid motifs within the MTS domain may be more important than the overall sequence similarities for mitochondrial targeting. PMID- 12527431 TI - Mutation of a putative nuclear receptor binding site abolishes activity of the nestin midbrain enhancer. AB - Regional differences in gene expression are critical to the proper development of specialized cell types in the nervous system. The ventral midbrain is the prominent source of dopaminergic neurons, which are affected in Parkinson's disease. We have recently identified a gene regulatory element that is specifically active in ventral midbrain neuroepithelium of developing embryos. This 204-bp transcriptional enhancer is conserved within the second intron of mammalian nestin genes and contains a putative binding site for a protein of the nuclear receptor family. Our present study shows, by mutagenesis and reporter gene assay in transgenic mice, that this site is essential for enhancer function in the developing midbrain. The characterization of regulatory sites and transcription factors with specific activity in the ventral midbrain provides insight into the molecular mechanisms by which neural progenitor cells become specified towards particular neuronal differentiation pathways. PMID- 12527432 TI - Characterization of a novel human breast cancer associated gene (BCA3) encoding an alternatively spliced proline-rich protein. AB - As part of an integrated study of breast cancer gene expression, partial cDNAs were cloned from normal and tumor breast cells by subtractive-hybridization and differential display cloning. The DNA sequence for one of these breast cancer associated genes was used to construct the larger 1319 bp BCA3 cDNA sequence using ESTs without assigned names or functions. High-level BCA3 mRNA expression was found in breast and prostate tumor cell lines whereas normal breast and prostate tissues have low-level expression. Further analysis revealed possible functional domains and alternative splicing of BCA3 that we confirmed by RT-PCR analysis. Immunohistochemistry revealed that the protein is expressed in breast tumor cells in vivo, and not in surrounding stromal tissue. PMID- 12527434 TI - The history of research in foot-and-mouth disease. AB - The history of research in foot-and-mouth disease falls into several distinct areas. In this short chapter I have highlighted what I consider to be the significant advances in our knowledge of the disease and its causal agent. 1. Loeffler and Frosch's landmark description in 1898 that the disease is caused by a filterable agent, the first observation that an animal disease could be caused by a virus. 2. The search for experimental laboratory animals, culminating in the demonstration by Waldmann and Pape of the susceptibility of the guinea pig in 1920 and the suckling mouse by Skinner in 1951. 3. The discovery of three distinct serotypes O, A and C in the 1920s by Vallee and Carre in France and by Waldmann in Germany, and the subsequent recognition in the 1940s and 1950s by the Pirbright group of the three Southern African Territory Types SAT 1-3, and Asia 1. 4. The development of in vitro techniques for the growth of the virus which have been crucial for the large-scale production of vaccines and for the accurate assay of virus infectivity. Early work by Hecke and the Maitlands in the early 1930s, followed by the crucial demonstration by Frenkel in 1947 that large amounts of the virus could be produced in surviving tongue epithelium, formed the basis for the vaccination programmes initiated in Europe in the 1950s. The subsequent development of cell lines has brought a remarkable degree of sophistication to the study of virus growth. 5. The impact of molecular studies on the structure of the virus and its mode of replication which have led to practical applications such as an in vitro test for vaccine potency, rapid diagnosis methods, and international epidemiological surveys. In addition, they have provided the means to design molecular vaccines. PMID- 12527435 TI - Molecular basis of pathogenesis of FMDV. AB - Current understanding of the molecular basis of pathogenesis of foot-and-mouth disease (FMD) has been achieved through over 100 years of study into the biology of the etiologic agent, FMDV. Over the last 40 years, classical biochemical and physical analyses of FMDV grown in cell culture have helped to reveal the structure and function of the viral proteins, while knowledge gained by the study of the virus' genetic diversity has helped define structures that are essential for replication and production of disease. More recently, the availability of genetic engineering methodology has permitted the direct testing of hypotheses formulated concerning the role of individual RNA structures, coding regions and polypeptides in viral replication and disease. All of these approaches have been aided by the simultaneous study of other picornavirus pathogens of animals and man, most notably poliovirus. Although many questions of how FMDV causes its devastating disease remain, the following review provides a summary of the current state of knowledge into the molecular basis of the virus' interaction with its host that produces one of the most contagious and frightening diseases of animals or man. PMID- 12527436 TI - Structure and receptor binding. PMID- 12527437 TI - Evolution of foot-and-mouth disease virus. AB - Foot-and-mouth disease virus evolution is strongly influenced by high mutation rates and a quasispecies dynamics. Mutant swarms are subjected to positive selection, negative selection and random drift of genomes. Adaptation is the result of selective amplification of subpopulations of genomes. The extent of adaptation to a given environment is quantified by a relative fitness value. Fitness values depend on the virus and its physical and biological environment. Generally, infections involving large population passages result in fitness gain and population bottlenecks lead to fitness loss. Very different types of mutations tend to accumulate in the foot-and-mouth disease virus (FMDV) genome depending on the virus population size during replication. Quasispecies dynamics predict higher probability of success of antiviral strategies based on multivalent vaccines and combination therapy, and this has been supported by clinical and veterinary practice. Quasispecies suggest also new antiviral strategies based on virus entry into error catastrophe, and such procedures are under investigation. Studies with FMDV have contributed to the understanding of quasispecies dynamics and some of its biological implications. PMID- 12527438 TI - Molecular epidemiology of foot-and-mouth disease virus. AB - Foot-and-mouth disease (FMD) is the most economically important veterinary pathogen due to its highly infectious nature, ability to cause persistent infections and long term effects on the condition and productivity of the many animal species it affects. Countries which have the disease have many trade restrictions placed upon them. In the last 15 years there have been significant advances in the understanding of FMD epidemiology. These have largely been due to the application of the molecular biological techniques of polymerase chain reaction amplification and nucleotide sequencing. In the World Reference Laboratory for FMD (Pirbright, UK), a large sequence database has been built up. This database has been used to aid in the global tracing of virus movements. It has been possible to genetically group many FMDV's based on their geographic origin and this has led to their being referred to as topotypes. The implications of this are that inter-regional spread of viruses can often be easily recognised and any evolutionary changes which subsequently occur can be monitored. Using these techniques, for the first time, we have been able to unequivocally show the recent pandemic spread of a FMDV type O strain through the whole of Asia and into Africa and Europe. This type of surveillance will become increasingly important as further globalisation of markets occurs. An increased understanding of how FMDV strains move between geographic regions will play a pivotal role in the development of future disease control strategies. PMID- 12527439 TI - FMD vaccines. PMID- 12527440 TI - Control and eradication of foot-and-mouth disease. PMID- 12527441 TI - Foot and mouth disease in wildlife. PMID- 12527442 TI - On the origins of autobiographical memory. AB - Tolving argues that one form of explicit memory, autobiographical memory is uniquely human and has no nonhuman animal antecedents. We suggest that a form of memory used by humans and nonhuman animals, dead reckoning, shares a common limbic structure, including the cingulate cortex and hippocampus, and involves similar processes in recognition of self-action. Thus, it may be homologous to, and an antecedent of, autobiographical memory in humans. PMID- 12527443 TI - Behavioral comparison of 4 and 6 month-old Ts65Dn mice: age-related impairments in working and reference memory. AB - Ts65Dn mice are partially trisomic for a segment of murine chromosome 16 similar to the gene segment on human chromosome 21 affected in Down's syndrome (DS). These animals display cognitive deficits, neurochemical imbalances, and cholinergic degeneration resembling alterations in DS and early onset Alzheimer's disease. The loss of basal forebrain cholinergic phenotype in Ts65Dn mice begins at approximately 6 months of age and may be due to an improperly functioning neurotrophic system. We compared 4 and 6 month-old Ts65Dn mice in a water-escape radial-arm maze task to investigate working and reference memory before and after the reported onset of cholinergic decline. Both 4 and 6 month-old Ts65Dn mice exhibited impaired performance compared to age-matched controls. However, the younger Ts65Dn mice displayed the capability to learn all working and reference memory measures, while the older Ts65Dn mice did not. Ts65Dn mice failed to maintain performance as working memory load increased, and the ability to handle an increasing working memory load also diminished with age. Collectively, these data suggest that major alterations in cognitive function occur in Ts65Dn mice between the ages of 4 and 6 months. PMID- 12527444 TI - Overexpression of an epitope-tagged serotonin transporter in serotonin neurons of the dorsal raphe nucleus using a defective HSV-1 vector. AB - The serotonin transporter (5HTT) plays a central role in serotonin neurotransmission. Abnormalities of 5HTT function have been implicated in depression, anxiety and alcohol intake. To better understand the functional role of this important molecule, we have utilized a viral vector approach to overexpress the 5HTT in regions of the rat brain. We have constructed a bicistronic defective herpes virus (HSV-1) vector that expresses both an epitope tagged 5HTT as well as beta-galactosidase (beta-GAL) as a marker for infected cells. The vector was capable of conferring serotonin uptake activity to Vero cells in culture, indicating transfer of a functional 5HTT. Injection of the 5HTT virus into the rat brain resulted in a dense focus of specific 125I RTI-55 binding at the injection site, indicating that the virally expressed 5HTT can also bind ligand when expressed in the brain. We were also able to overexpress an epitope tagged 5HTT in serotonergic neurons in the dorsal raphe nucleus (DRN) using this approach. These data demonstrate that the levels of the 5HTT in 5HT neurons can be manipulated in the adult rodent brain using an HSV-1 vector. PMID- 12527445 TI - S22153, a melatonin antagonist, dissociates different aspects of photoperiodic responses in Syrian hamsters. AB - In the Syrian hamster, short photoperiod (SP) induces changes in several physiological functions (body mass, reproduction, hibernation), and these responses involve the pineal hormone melatonin. The present study investigated the effects of a melatonin antagonist, S22153, on photoperiodic adaptation of male Syrian hamster. When constantly released from subcutaneous implants, S22153 had no effect on body or testes masses of animals kept in long photoperiod. S22153 decreased the total hibernation duration observed in animals exposed to SP and low temperature. The decrease in hibernation duration was due to a marked reduction in the number and duration of hypothermic bouts. Moreover, S22153 significantly inhibited the increase of interscapular brown adipose tissue (BAT) mass induced by SP. However, neither the gonadal atrophy nor the body mass increase induced by SP were affected by S22153. These results show that S22153 affects only part of the physiological changes controlled by SP and cold. Whether the decreases in BAT mass and hibernation duration are linked still remains an open question. PMID- 12527446 TI - Effects of intra-accumbens focal administrations of glutamate antagonists on object recognition memory in mice. AB - Generally recognition memory is distinguished into spatial and object memories that have been suggested to relay at a cortical level on different neural substrates. Recent studies point to a possible involvement of the nucleus accumbens (Nac) in spatial memory, demonstrating that blockade of glutamate antagonists within this structure impairs acquisition and consolidation of spatial information, while not many data are available on the potential role of this structure in object recognition. Thus in this study we wanted to investigate the effects of intra-accumbens focal administrations of NMDA antagonist, AP-5 (0.05, 0.1, 0.15 or 0.2 microg per side), and AMPA antagonist, DNQX (0.0005 or 0.001 microg per side), in object recognition memory. The spontaneous preference displayed by mice for novel objects was taken as an index for measuring object recognition. Pre-training focal administrations of both antagonists impaired the ability of mice to selectively explore the novel object in test session. However, the AMPA antagonist induced also a decrease in exploration and locomotion. In order to assess whether glutamate receptors located within the Nac were also involved in subsequent steps of object information processing, we performed additional experiments injecting AP-5 and DNQX immediately after training and testing the animals 24-h later. In this case, AP-5 but not the AMPA antagonist impaired exploration of the novel object. These results demonstrate that the Nac is involved in object recognition, and confirm that the different glutamate receptors mediate different component of information processing within the accumbens. PMID- 12527447 TI - Effects of light deprivation on recovery from neglect and extinction induced by unilateral lesions of the medial agranular cortex and dorsocentral striatum. AB - A number of previous studies have indicated that an environmental manipulation, 48 h of light deprivation (LD), produces virtually complete and permanent behavioral recovery of function from neglect induced by medial agranular cortex (AGm) lesions. LD-induced behavioral recovery from neglect is correlated with physiological changes in the dorsolateral striatum, an area that contains the projection zone of AGm efferents in the dorsocentral striatum (DCS). In this study, the behavioral effects of 48 h of LD on subjects with either unilateral DCS, AGm, or combined AGm/DCS lesions were investigated to examine whether the integrity of the DCS is crucial for behavioral recovery from neglect and whether LD will have a therapeutic effect on extinction deficits. Subjects were tested for extinction to bilateral simultaneous stimulation of the forepaws, and visual, auditory and tactile neglect. Forty-eight hours of LD failed to produce behavioral recovery from neglect in rats with DCS lesions, or a therapeutic affect on extinction deficits in any of the groups. The results of this study further support the crucial role of the DCS in recovery from neglect induced by AGm lesions and suggests that the DCS may be the crucial site for the mechanisms leading to LD-induced recovery. Further, the ineffectiveness of LD on extinction suggests that components of the neglect syndrome are dissociable and may require different therapeutic interventions. PMID- 12527448 TI - Dorsal raphe nucleus modulates neuronal activity in rat intergeniculate leaflet. AB - Serotonergic input from midbrain raphe nuclei is believed to have a significant effect on mammalian circadian timing system. The suprachiasmatic nucleus (SCN) receives its serotonergic input from the median raphe nucleus, while the intergeniculate leaflet (IGL) receives serotonergic innervation from the dorsal raphe nucleus (DRN). The present paper was aimed at determining whether projection from the DRN affected rhythmic neuronal oscillations in the IGL of rats. We investigated the impact of electrolytic lesions and electric stimulation of the DRN on spontaneous isoperiodic (i.e. burst firing with a constant interburst interval) neuronal activity recorded in the IGL. In all our experiments a complete lesion of the DRN always caused a significant increase (ca. 100%) of spontaneous activity of IGL neurons, their oscillatory character having been maintained, though. On the other hand, electric stimulation of the DRN produced a transient decrease in firing rate oscillations of the IGL neurons. The obtained results indicate that the neuronal projection from the DRN has a substantial modulating effect on IGL activity-an important element of the mechanism of the circadian time-keeping system that mediates the transfer of non photic information to the SCN by modulating its activity. The observed increase of isoperiodic activity in the IGL after DRN lesion and a transient decrease in this activity after electric stimulation indicate an inhibitory character of this effect. The present findings corroborate the hypothesis that the DRN is a one of the major and extremely important source of the modulatory inputs to the mammalian circadian time-keeping system. PMID- 12527449 TI - Behavioral deficits and recovery following transient focal cerebral ischemia in rats: glutamatergic and GABAergic receptor densities. AB - The neurobiologic mechanisms underlying the recovery process following stroke are poorly understood. The present study investigated glutamatergic and gamma-amino butyric acid (GABA)-ergic receptor densities following experimental stroke in rats exposed to different environmental housing or pharmacologic interventions. About 2 days after transient (120 min) middle cerebral artery (MCA) occlusion, the rats were singly housed in standard cages or were moved to an enriched environment and treated for 10 days with either 0.9% NaCl or with the alpha(2) adrenoceptor antagonist, atipamezole (1.0 mg/kg, s.c.). The limb-placing, foot slip, and water-maze tests were used to assess behavioral deficits and recovery following ischemia. The rats were decapitated on day 25 after the operation and their brains were processed for [3H]MK-801, [3H]D,L,-alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid (AMPA), [3H]kainate, and [3H]muscimol autoradiography. Receptor binding site densities were different between sham operated rats and ischemic rats only in the lesion core and lateral ventroposterior thalamic nucleus. Ischemic rats housed in an enriched environment and treated with atipamezole had better performance in the limb-placing test. The deficit in the water-maze test was most pronounced in ischemic rats housed in standard cages. There were a number of correlations between the behavioral data and receptor binding densities in ischemic rats. For example, recovery in the limb-placing test correlated with [3H]AMPA receptor binding sites in the contralateral frontal cortex (r=0.616, P<0.05), hindlimb cortex (r=0.649, P<0.05), and parietal cortex (r=0.674, P<0.05) in ischemic rats housed in an enriched environment. There were similar correlations between limb-placing recovery and [3H]kainate binding sites in the contralateral cortices in ischemic rats housed in standard cages. In addition, there were particularly strong clustered correlations between swimming speed in the water-maze test and [3H]AMPA receptor binding sites in the hippocampal subregions in the ischemic rats housed in an enriched environment. The present results suggest that transient focal cerebral ischemia does not induce significant long-term changes in glutamatergic and GABAergic receptors in areas remote from the infarct area. The correlational data, however, suggest an important role for the contralateral cortex in the behavioral outcome and maintenance of the recovered state of ischemic rats, depending on housing conditions. In addition, attenuation of spatial learning deficits observed in ischemic rats housed in an enriched environment might be due to an increase in the swimming speed through hippocampal AMPA receptor-mediated mechanisms. PMID- 12527450 TI - Concanavalin A application to the olfactory epithelium reveals different sensory neuron populations for the odour pair D- and L-carvone. AB - Carvone enantiomers (D and L optical isomers) have been shown to be discriminable by humans even though the odor qualities are quite similar. Our experiment is based on a finding (J. Steroid Biochem. Molec. Biol. 1991;39(4B):621) that Concanavalin A (ConA) applied to a frog olfactory epithelium preparation blocks cAMP transduction induced by D- but not by L-carvone. We used standard operant conditioning methods to train animals to discriminate low odor concentrations of D-carvone from clean air, to discriminate L-carvone from clean air; or to discriminate between clean air and the odors of D-carvone, L-carvone, ethyl acetate and methacrylic acid. After perfusion of the nasal cavity with ConA, rats did not respond to D-carvone above or near chance level, while the L-carvone response was not affected at the same or higher ConA doses. However, for rats trained on both enantiomers and the two other unrelated odorants, the D-carvone response remained unaffected by ConA. These results suggest to us that: (1) ConA blocks at least one chiral receptor selective for D-carvone; (2) D-carvone odor quality is modified by ConA so that it is no longer recognized by rats trained on D-carvone only, while rats trained to generalize odors still respond to D carvone. PMID- 12527451 TI - The corticotropin-releasing factor receptor 1 antagonist CP-154,526 reverses stress-induced learning deficits in mice. AB - The neuropeptide corticotropin-releasing factor (CRF) coordinates the endocrine responses to stress as a major physiological regulator of the hypothalamic pituitary-adrenal axis. We assessed the effect of the non-peptidergic CRF receptor 1 antagonist CP-154,526 on stress-induced changes in context-dependent fear conditioning and hippocampal synaptic plasticity. The learning impairment of mice trained immediately after 1 h immobilization could be overcome by preinjection of CP-154,526 before exposure to immobilization. Exposure to acute stress reduced the amount of autophosphorylated Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) in the hippocampal CA1 area. When animals were pretreated with CP-154,526 before immobilization, the amount of hippocampal autophosphorylated CaMKII was elevated. Electrophysiological studies in the hippocampal CA1 region of stressed animals revealed no significant effects of the CP-154,526 pretreatment on long-term potentiation but a significant elevation of paired-pulse facilitation (PPF) was observed. The CP-154,526-induced enhancements in fear conditioning and PPF could be prevented by the selective CaMKII inhibitor KN-62. Our results demonstrated that learning impairment after acute stress was antagonized by CP-154,526 pretreatment. PMID- 12527453 TI - An editorial welcome to the new year 2003. PMID- 12527452 TI - Spatial, contextual and working memory are not affected by the absence of mossy fiber long-term potentiation and depression. AB - The mossy fibers of the hippocampus display NMDA-receptor independent long-term plasticity. A number of studies addressed the role of mossy fiber long-term plasticity in memory, but have provided contrasting results. Here, we have exploited a genetic model, the rab3A null-mutant, which is characterized by the absence of both mossy fiber long-term potentiation and long-term depression. This mutant was backcrossed to 129S3/SvImJ and C57Bl/6J to obtain standardized genetic backgrounds. Spatial working memory, assessed in the eight-arm radial maze, was unchanged in rab3A null-mutants. Moreover, one-trial cued and contextual fear conditioning was normal. Long-term spatial memory was tested in the Morris water maze. Two different versions of this task were used, an 'easy' version and a 'difficult' one. On both versions, no differences in search time and quadrant preferences were observed. Thus, despite the elimination of mossy fiber long-term plasticity, these tests revealed no impairments in mnemonic capabilities. We conclude that spatial, contextual and working memory do not depend on mossy fiber plasticity. PMID- 12527454 TI - Maternal and perinatal outcome in rheumatic heart disease. AB - OBJECTIVES: To study the maternal and perinatal outcome of pregnancies complicated by rheumatic heart disease. METHODS: A retrospective study was carried out in the cardio-obstetric clinic of the Postgraduate Institute of Medical Education and Research, Chandigarh (India) over a period of 13 years (1987-1999) involving 486 pregnant patients with rheumatic heart disease. Maternal and perinatal outcome was reviewed. RESULTS: Three hundred and four patients (63.3%) had single valve involvement and mitral stenosis was the most predominant lesion (89.2%). One hundred and seventy one (38.6%) patients had undergone surgical correction prior to the onset of pregnancy. One hundred and thirteen patients (22.6%) were identified as NYHA class III-IV. Mitral valvotomy was performed during pregnancy in 48 patients. The incidence of preterm birth and small for gestational age newborns was 12% and 18.2%, respectively. There were 10 maternal deaths, of which eight patients were NYHA III and IV. CONCLUSIONS: Rheumatic heart disease in pregnancy is associated with significant maternal and perinatal morbidity in NYHA class III-IV patients. PMID- 12527455 TI - The outcome of macrosomic fetuses in a low risk primigravid population. AB - OBJECTIVES: To ascertain whether fetal macrosomia is associated with increased maternal and neonatal morbidity in uncomplicated, singleton, vertex deliveries at term in primigravid women. METHODS: This was a retrospective population based survey of 8617 deliveries over an 11-year period. These were stratified into three birthweight categories: 2500-3999 g (n=7854), 4000-4499 g (n=666) and > or =4500 g (n=97). Outcome variables included maternal characteristics, delivery details, maternal and perinatal morbidity data. RESULTS: Increased BMI and incidence of Caucasian ethnicity and non-smoking were significantly greater in macrosomic compared with non-macrosomic infants (P<0.001). Increasing birthweight (especially > or =4.5 kg) was associated with significantly (P<0.001) lower rates of spontaneous onset of labor, spontaneous vertex deliveries and significantly higher rates (P<0.001) of maternal and neonatal morbidity. CONCLUSIONS: This study supports the notion of expectant management in suspected fetal macrosomia in low risk primigravid women until 40 weeks gestation. Thereafter, the safest mode of delivery is controversial, with some evidence pointing to elective cesarean section as a viable alternative in these women. A prospective RCT is needed to evaluate the best management option in terms of fetal and maternal outcome in cases of suspected macrosomia after 40 weeks' gestation. PMID- 12527456 TI - Intracervical misoprostol and prostaglandin E2 for labor induction. AB - OBJECTIVES: To compare the safety and efficacy of misoprostol with PGE(2) for induction of labor by intracervical administration. METHODS: Eighty-six women with indications for labor induction at term were randomly assigned to two groups. Each woman received either 50 microg of misoprostol or 0.5 mg of prostaglandin E(2) intracervically. If labor was not initiated after 4 h, the same dose was repeated every 4 h to a maximum of 200 microg of misoprostol or 1.5 mg of PGE(2) until adequate labor was achieved. RESULTS: Forty-three women were allocated to the misoprostol group and 43 to the prostaglandin E(2) group. Misoprostol was more effective than PGE(2) in producing cervical changes (P<0.025). Delivery within 12 h after the first administration occurred more often in the misoprostol group than in the PGE(2) one (85% vs. 56%, P<0.05). Less patients in the misoprostol group required oxytocin augmentation than in the PGE(2) one (16.3% vs. 39.5%, P<0.05). Uterine tachysystole and hyperstimulation occurred more frequently in the misoprostol group (44.1%) than in the PGE(2) group (18.7%) (P<0.05). Nevertheless, no statistically significant differences were noted between the two groups including mode of delivery and neonatal or maternal adverse outcome. The interval from induction to vaginal delivery was significantly shorter in the misoprostol group (480+/-172 min vs. 657+/-436 min, P<0.01). CONCLUSIONS: Compared with prostaglandin E(2), intracervical misoprostol is more effective in cervical ripening and labor induction at term. The higher frequency of uterine hypercontractility associated with the use of misoprostol did not increase the risk of adverse intrapartum and neonatal outcomes. PMID- 12527457 TI - Myocardial hypertrophy of the recipient twins in twin-to-twin transfusion syndrome and cerebral palsy. AB - OBJECTIVES: This study was performed to determine risk factors for cerebral palsy (CP) in monochorionic twins, especially with twin-to-twin transfusion syndrome (TTTS). METHODS: In 33 pathologically confirmed monochorionic pregnancies, we analyzed the incidence of cardiovascular and neurological complications. RESULTS: Seventeen cases were diagnosed as TTTS. Myocardial hypertrophy was detected in seven recipient twins of TTTS, but not found in the donor twins of TTTS or in non TTTS. CP developed in six out of 29 in TTTS and one out of 32 in non-TTTS. In TTTS, all the cases with CP were seen in the recipient twins. The development of CP was significantly associated with cardiovascular complications such as myocardial hypertrophy and hydropic changes. CONCLUSIONS: Myocardial hypertrophy found in the recipient twins in TTTS seems to be a risk for developing CP later on, which may have implications in understanding the pathogenesis of CP. PMID- 12527458 TI - Estrogen levels and estrogen receptors in patients with stress urinary incontinence and pelvic organ prolapse. AB - OBJECTIVES: To investigate the histologic characteristics of tissues presumed to be the cause of urinary stress incontinence and pelvic organ prolapse. METHODS: Cardinal ligament and uterosacral ligament samples were obtained from 73 women undergoing hysterectomy. The evaluation of estrogen receptors (ERs) by immunohistochemical staining was semi-quantitative. Serum estrogen was determined by ELISA. Statistical analyses were performed by the independent-sample t-test and one-way ANOVA. RESULTS: Serum estradiol levels and ER values in the premenopausal women with pelvic organ prolapse were significantly lower than in the control group (P<0.01). A positive correlation was found between ERs and the number of postmenopausal years (P<0.01). ER values were similar in the cardinal and uterosacral ligaments. CONCLUSIONS: Serum estrogen levels and ER values are significantly lower in the uterine ligaments of premenopausal women with pelvic organ prolapse, and there was a positive correlation between ER values in the uterine ligaments and the duration of postmenopausal years. Serum estrogen levels and ER values were similar in the cardinal ligament and the uterosacral ligament. PMID- 12527459 TI - Risk factors as the basis for triage in low-resource cervical cancer screening programs. AB - OBJECTIVES: Several studies have shown impressive sensitivity and specificity of visual inspection using acetic acid wash (VIA) in detecting cervical dysplasia. This study examines how risk-factor based triage, using clinical history-taking to limit the testing to those at highest risk of disease, improves the clinical utility of VIA. METHODS: The study population included 2206 woman aged 25-56 from peri-urban primary care clinics in Zimbabwe. Three risk factors variables [age, lifetime number of sexual partners and history of a sexually transmitted infection (STI)] were selected as the basis for computer-simulated patient triage. Criteria for selecting risk factors were biological relevance, programmatic feasibility, historical evidence in the scientific literature as a risk factor for cervical cancer and a significant (P<0.05) predictor of dysplasia in this study population. Predictive values for VIA were calculated on subgroups of women with combinations of the three risk factors and then compared with the predictive value of VIA calculated on the full study population. RESULTS: The positive predictive value (PPV) among women with all three risk factors (30.2%) was 1.6 times higher than that of all study women and 4.5 times higher than women with none of the three risk factors (4.1%). Additionally, the PPV was slightly (30%) higher than that obtained from computer-simulated serial testing using VIA as the primary test followed by HPV testing (27.4%). CONCLUSION: Clinical history taking could provide the basis for patient triage to increase the clinical utility of VIA. Such an approach could also serve to prioritize who gets tested first in low-resource countries with high disease prevalence that are struggling to strengthen national cervical cancer prevention programs. PMID- 12527460 TI - Effect of clomiphene citrate on endometrium thickness and echogenic patterns. AB - OBJECTIVE: To study the effect of clomiphene citrate (CC) on the thickness and echogenic pattern of the endometrium using vaginal sonography. METHOD: In this experimental prospective study, CC was given to 31 women with unexplained infertility. Thickness and echo patterns of the endometrium, as well as estradiol (E2) and progesterone (P) levels, were compared in women taking CC and in control patients during both the late proliferative and midsecretory phases of the menstrual cycle. RESULTS: Endometrial thickness was significantly thinner in women taking CC (P<0.03) than in controls during the late proliferative phase but there was no significant difference during the midsecretory phase. The endometrial echogenic patterns of women taking CC who had conceived revealed endometrial thicknesses of grade III and grade IV on midsecretory days, but these findings were not significantly different from those of women who had not conceived (P=0.3). Serum E2 level was higher in women taking CC than in the controls on both late proliferative and midsecretory days (P<0.05). CONCLUSION: CC affects endometrium thickness on late proliferative days but not on midsecretory days, and does not alter the echogenic pattern of the endometrium. PMID- 12527461 TI - Successful pregnancy after conservative management of bilateral ovarian abscesses. PMID- 12527462 TI - Intravenous leiomyomatosis extending to the heart. PMID- 12527463 TI - Misoprostol for second trimester termination of pregnancies with prior low transverse cesarean section. PMID- 12527464 TI - Risk factors for cervical cancer in Mozambican women. PMID- 12527465 TI - Intrauterine misoprostol for refractory postpartum hemorrhage. PMID- 12527466 TI - Polymorphism of codon 72 of p53 and environmental factors in the development of cervical cancer. PMID- 12527467 TI - Birthweight-specific neonatal mortality in developing countries and obstetric practices. AB - OBJECTIVES: To evaluate birthweight-specific neonatal mortality and perinatal interventions in major medical centers in developed and developing countries. METHODS: A survey was developed and electronically mailed to 13 medical centers participating in the Global Network for Perinatal and Reproductive Health (GNPRH). The ability of a center to provide requested data was assessed. The mortality rates and use of specific perinatal interventions in centers in developing countries were compared with developed countries. RESULTS: Nine centers in developing countries responded to the survey, and three centers in developed countries were used for comparison. Data collection was highly variable. Most developing country centers were able to provide data by birthweight but not by gestational age. The differences in mortality rates between developing and developed countries were more pronounced at lower gestational ages and birthweights. A difference was found in perinatal interventions between developing and developed countries. In the former, viability was generally considered 28 weeks, and the gestational age at which cesarean sections were usually performed for the sake of the fetus at preterm gestations varied from 26 to 37 weeks. Most centers did not routinely induce for pPROM; only five out of nine centers used antibiotics to prolong latency. Most centers used tocolysis beginning at 26-28 weeks through 32-37 weeks, and a variety of tocolytic agents were used. Most centers routinely used corticosteroids for preterm infants, and all centers employed repeat weekly steroid dosing if undelivered. CONCLUSIONS: Despite the fact that the GNPRH centers included in this study represent some of the best health care available in these countries, they lag far behind centers in developed countries in neonatal mortality rates and their use of various obstetric practices. Furthermore, incomplete and inconsistent data collection complicates the evaluation of the factors contributing to high neonatal mortality rates. PMID- 12527468 TI - Challenges of ethical research in resource-poor settings. AB - The disparity in resources between economically developed and developing countries presents ethical concerns when commercial sponsors of studies and investigators propose to conduct them with subjects in developing countries. Vulnerable people may be recruited into studies of little health benefit to themselves or their communities and, under undue inducements, may accept disproportionate risks. Reproductive health studies may present women with undue risks. Guidelines have been developed to protect exploitable populations in resource-poor settings, although guarding their right to make informed and voluntary choices poses special challenges. Guidelines pay special attention to pregnant women as research subjects, and may approve and even require their enrollment in studies of products not known to be harmful. Placebo-controlled studies are addressed in contexts where no recognized treatments are routinely accessible. The structure and functions of research ethics review committees present difficult challenges, but they may be mitigated by enlightened international collaboration. PMID- 12527469 TI - The mGluR5 antagonist MPEP, but not the mGluR2/3 agonist LY314582, augments PCP effects on prepulse inhibition and locomotor activity. AB - Phencyclidine (PCP), a non-competitive antagonist of ionotropic N-methyl-D aspartate (NMDA) receptors, produces psychotomimetic effects, such as a disruption in prepulse inhibition (PPI) of the startle response. NMDA antagonists also induce locomotor hyperactivity in rodents. We hypothesized that, like NMDA receptors, metabotropic glutamate receptors (mGluRs) modulate PPI and locomotor activity either alone or, in the case of mGluR5, via interaction with NMDA receptors. Rats treated with the mGluR5 antagonist MPEP (2-methyl-6 phenylethynylpyridine) or the mGluR2/3 agonist LY314582, either alone or in combination with PCP, were tested in PPI and locomotor activity paradigms. Neither MPEP nor LY314582 altered PPI. MPEP, but not LY314582, potentiated the PPI-disruptive effects of PCP. MPEP alone did not alter locomotor or exploratory behavior, but augmented the complex, time-dependent locomotor-stimulating effects of PCP. LY314582 dose-dependently decreased locomotor activity and exploratory holepokes. LY314582 did not alter the PCP-induced increases in locomotor activity, but further decreased the number of holepokes. The effects of MPEP on the response to PCP may reflect the cooperation and co-localization of NMDA and mGlu5 receptors. PMID- 12527470 TI - Morphine conditioned reward is inhibited by MPEP, the mGluR5 antagonist. AB - In the present study we examined the effect of MPEP [2-methyl-6-(phenylethynyl) pyridine] a potent, selective and systemically active metabotropic glutamate receptor (mGluR) type I (subtype mGluR5) antagonist on conditioned morphine reward in mice. In an unbiased version of conditioned place preference (CPP) paradigm, single conditioning with 10 mg/kg of morphine produced reliable place preference. MPEP at 30, but not 10 mg/kg significantly inhibited the acquisition as well as expression of morphine-induced CPP, but it neither produced place preference or aversion, nor affected locomotor activity of mice. Effects of MPEP on learning and memory were studied in the elevated plus maze model of spatial learning. In contrast to 0.1 mg/kg of MK-801, which inhibited the acquisition of this task, 30 mg/kg of MPEP affected neither learning nor memory retrieval. These data suggest that mGluR5 may be involved in conditioned morphine reward. PMID- 12527471 TI - Extracellular glutamate levels in prefrontal cortex during the expression of associative responses to cocaine related stimuli. AB - We conducted studies to examine the potential role of glutamate in the prefrontal cortex (PFC) during conditioned responses to stimuli (flashing light and metronome) previously associated with cocaine administration. During training, PAIRED subjects received cocaine injections (15 mg/kg) during stimuli sessions while UNPAIRED subjects received saline injections (but received cocaine in the home cage an hour later). We showed previously that PAIRED subjects exhibit conditioned locomotion when tested with the stimuli alone. In this study, we further demonstrated that the expression of behavioral sensitization in response to cocaine challenge is under conditioned control in PAIRED subjects. Then, we used microdialysis to examine extracellular levels of glutamate in the PFC in response to presentation of the conditioned stimuli alone and challenge with cocaine in the presence of the conditioned stimuli. Although PAIRED subjects demonstrated conditioned locomotion and conditioned control of sensitization during the microdialysis experiment, PFC glutamate levels were unaltered during the tests and did not differ between the PAIRED and UNPAIRED subjects. PMID- 12527472 TI - Chronic antidepressant treatment increases the membrane expression of AMPA receptors in rat hippocampus. AB - It has been proposed that potentiation of AMPA receptor (AMPAR) function may be useful in the treatment of depression. Here we studied the acute and chronic effect of the antidepressants desipramine and paroxetine, which differentially affect monoamine reuptake, on the expression of the AMPAR subunits GluR1 and GluR2/3, analyzed by Western blot, both in total and in membrane-enriched extracts from rat hippocampus. Acute antidepressant treatment did not produce any change in the expression of AMPAR subunits. In chronic treatments, rats were daily treated with the antidepressants (10 mg/kg/day) for 7, 14, or 21 days. In rats receiving either of the two antidepressant treatments for 21 consecutive days and killed 24 h after the last injection, an increase in GluR1 and GluR2/3 levels was found in the membrane fraction, with no significant change in the total extract, suggesting a trafficking of the AMPAR subunits from intracellular pools to synaptic sites in the hippocampus. This appeared to be a region-specific effect since no change in AMPAR subunit expression was found in the frontal cortex. Previously reported modifications in phosphorylating enzymes by chronic antidepressants could perhaps play a role in hippocampal membrane insertion of AMPAR subunits. When the survival time after the 21-day-treatment was longer - 72 instead of 24 h - the hippocampal membrane expression of GluR1, but not of GluR2/3 subunits, was still increased, as could be expected from the distinct mechanisms operating in synaptic delivery of GluR1 and GluR2/3 subunits. The antidepressant-induced increase in the number of GluR1- and GluR2/3-containing AMPARs at the synapses may indicate an enhanced AMPAR-mediated synaptic transmission which could help to counteract the alterations in neuronal connectivity which appear to underlie the pathophysiology of mood disorders. PMID- 12527473 TI - Neuroanatomical substrates involved in the anxiogenic-like effect of acute fluoxetine treatment. AB - An initial exacerbation of anxiety can be observed in animals and humans treated with selective serotonin reuptake inhibitors (SSRIs). The neurobiological substrates and mechanism(s) underlying this effect are not clear. We used Fos expression as a marker of neuronal activation to investigate effects of acute fluoxetine treatment in rats submitted to two different models of emotional stress, airjet and immobilization. Exposure to both stressors induced Fos expression in various brain regions implicated in fear/anxiety mechanisms. Acute treatment with 5 mg/kg fluoxetine facilitated airjet-induced escape responses and enhanced the airjet-, as well as immobilization-induced Fos expression exclusively in the locus coeruleus (LC), but not in other areas including the amygdala, hypothalamus or septum. Fluoxetine also facilitated airjet-induced noradrenaline efflux in the medial prefrontal cortex, a projection area of LC noradrenergic neurons. A higher dose of fluoxetine (10 mg/kg) did not change escape responses and had no effect on stress-induced Fos expression in the LC, but decreased airjet-induced Fos expression in the medial amygdala. The results indicate that anxiogenic effects of acute fluoxetine treatment occur in a specific dose range and can be mimicked by exacerbation of escape responses in the airjet model. Furthermore, facilitation of escape responses by fluoxetine is linked to enhanced activity in the LC/noradrenaline system. PMID- 12527474 TI - Acute treatment with the cyclic antidepressant desipramine, but not fluoxetine, increases membrane-associated G protein-coupled receptor kinases 2/3 in rat brain. AB - The homologous regulation of receptors is mediated by G protein-coupled receptor kinases (GRKs) which phosphorylate the agonist-activated receptor. This study was designed to assess the in vivo indirect activation of adrenoceptors or 5-HT receptors by the reuptake blocker desipramine or fluoxetine on the cellular distribution of GRK 2/3 in rat brain. Immunoblot analysis (frontal cortex) with a GRK 2 antibody revealed a unique 80 kDa protein (mixed GRK 2/3) in total homogenate (H) and in membrane (P2) and cytosolic (S2) fractions. The proportion of GRK 2/3 in each fraction, relative to that of H, was: P2/H=0.11 and S2/H=0.45. Acute desipramine (noradrenaline reuptake blocker) increased in a dose- (1-30 mg/kg, i.p.) and time- (1-6 h) dependent manner the content of GRK 2/3 in the membrane (P2/H ratios increased by 37-164%). This effect vanished with a prolonged desipramine (30 mg/kg) exposure (24 h). Desipramine did not alter the content of GRK 2/3 in the cytosol (S2/H ratios). Chronic desipramine (10 mg/kg every 24 h) for 14 days did not change significantly the immunodensity of GRK 2/3 in the membrane or the cytosol. The acute administration (2 h) of fluoxetine (5 HT reuptake blocker; 3-30 mg/kg) did not induce significant changes in the content of GRK 2/3 in the membrane (P2/H ratio) or cytosol (S2/H ratio). The results indicate that the in vivo activation of adrenoceptors by desipramine is associated with a time-dependent modulation of membrane-associated GRK 2/3 (i.e. an acute increase in the kinase content which is followed by a return to the basal expression upon repeated treatment). In contrast, the acute in vivo activation of 5-HT receptors induced by fluoxetine does not result in modulation of GRK 2/3. PMID- 12527475 TI - Lack of self-administration and behavioural sensitisation to morphine, but not cocaine, in mice lacking NK1 receptors. AB - Mice lacking the NK1 receptor, the preferred receptor for substance P, demonstrate normal analgesic responses to morphine on the hot plate assay, but have been predicted, on the basis of conditioned place preference studies, to be insensitive to the rewarding properties of opiates. In this study, self administration and the development and maintenance of locomotor sensitisation of both morphine and cocaine were investigated in mice that lacked the NK1 gene (NK1 knockout mice, NK1(-/-)). Both wildtype and NK1(-/-) mice learned an operant lever-press response to obtain food. When intravenous infusions of morphine (0.2 mg/kg/infusion) were substituted for the food reward, the wildtype mice initially reduced rates of lever pressing, but then increased them on the rewarded lever to obtain approx. 10 infusions per 90 min session; in contrast, NK1(-/-) mice continued to operate both the rewarded, and non-rewarded levers at low rates. Additionally, NK1(-/-) mice failed, following repeated administration, to sensitise to the locomotor stimulant effects of morphine (15 mg/kg, i.p.). These deficits were specific to opiates, since NK1(-/-) mice responding for food or cocaine self-administration (0.65 mg/kg/infusion) did not differ from wildtypes, and they showed normal behavioural sensitisation to repeated cocaine administration (10 mg/kg, i.p.). These results demonstrate that NK1 receptors are critical for the reinforcing properties of morphine, and for adaptive responses elicited by repeated opiate administration. It is postulated that substance P and the NK1 receptor may be necessary for the development of opiate, but not cocaine addiction. PMID- 12527476 TI - Antidepressants reduce phosphoinositide-specific phospholipase C (PI-PLC) activity and the mRNA and protein expression of selective PLC beta 1 isozyme in rat brain. AB - In order to examine whether antidepressants mediate their action by interacting with one of the key components of the phosphoinositide (PI) signaling pathway, i.e. PI-specific phospholipase C (PLC), and whether this represents a common mechanism of action of antidepressants, we determined the effects of antidepressants of various classes on PI-PLC activity and on the expression of PLC isozymes in rat brain. It was observed that chronic (21-day) but not acute (1 day) administration with desipramine (DMI), fluoxetine (FLX) and phenelzine (PHLZ), decreased PI-PLC activity in membrane and cytosol fractions of cortex and hippocampus. Similar changes were observed with alprazolam (ALP) and buspirone (BUS), who possess anxiolytic and antidepressant properties. On the other hand, an anxiogenic drug, metachlorophenylpiperazine (MCPP), increased PI-PLC activity in both membrane and cytosol fractions of cortex and hippocampus. The immunolabeling studies showed that all the antidepressants and anxiolytics that caused a decrease in PI-PLC activity also selectively decreased the protein levels of a specific isozyme of PLC, i.e. PLCbeta(1), in membrane and cytosol fractions of cortex and hippocampus, whereas MCPP increased the levels of this particular isozyme. These changes were accompained with changes in the mRNA levels of PLCbeta(1), as determined by quantitative RT-PCR. These antidepressants and anxiolytics did not cause significant changes in the expression of PLC delta(1) or gamma(1) isozyme. Our results thus suggest that modulation of PI-PLC may be common to all classes of antidepressants, which in turn, may be associated with their mechanisms of action. PMID- 12527477 TI - The age-related discrepancy in the effect of neuropeptide Y on select catecholamine biosynthetic enzymes in the adrenal medulla and hypothalamus in rats. AB - The elevated levels of circulating catecholamines (CAs) with age may be related to the increased expression of CA biosynthetic enzymes, tyrosine hydroxylase (TH) and dopamine beta hydroxylase (DbetaH) in the adrenal medulla of senescent compared with younger animals. Neuropeptide Y (NPY) is co-synthesized and co released with CAs in the adrenal medulla. NPY inhibits the stimulated secretion of CAs, however, its role in regulation of the genes encoding CA biosynthetic enzymes is not clear. We hypothesized that NPY up-regulates TH, DbetaH and NPY expression in the adrenal medullae of young and old Fischer-344 rats. NPY increased mRNA expression of TH, DbetaH, NPY and also enhanced TH protein level in the adrenal medullae of young rats by 50%, 35%, 45% and by 20%, respectively. We also examined the effect of NPY on TH and NPY mRNA in the hypothalamus. Basal expression of TH mRNA was decreased in the hypothalamus with age. DNA binding activities of activator protein-1 and cAMP response element binding protein were also augmented only in the young by 140% and 125%, respectively. We conclude that NPY stimulates the CA biosynthetic pathway in the adrenal medulla and positive auto-regulation of NPY might be involved in this process. The stimulatory effect of NPY on adrenomedullary CA biosynthetic pathway is blunted with age. PMID- 12527478 TI - Gas6 rescues cortical neurons from amyloid beta protein-induced apoptosis. AB - Gas6, a product of the growth-arrest-specific gene 6, protects neurons from serum deprivation-induced apoptosis. Neuronal apoptosis is also caused by amyloid beta protein (Abeta), whose accumulation in the brain is a characteristic feature of Alzheimer's disease. Abeta induces Ca(2+) influx via L-type voltage-dependent calcium channels (L-VSCCs), leading to its neurotoxicity. In the present study, we investigated effects of Gas6 on Abeta-induced cell death in primary cultures of rat cortical neurons. Abeta caused neuronal cell death in a concentration- and time-dependent manner. Gas6 significantly prevented neurons from Abeta-induced cell death. Gas6 ameliorated Abeta-induced apoptotic features such as the condensation of chromatin and the fragmentation of DNA. Prior to cell death, Abeta increased influx of Ca(2+) into neurons through L-VSCCs. Gas6 significantly inhibited the Abeta-induced Ca(2+) influx. The inhibitor of L-VSCCs also suppressed Abeta-induced neuronal cell death. The present cortical cultures contained few non-neuronal cells, indicating that Gas6 affected the survival of neurons directly, but not indirectly via non-neuronal cells. In conclusion, we demonstrate that Gas6 rescues cortical neurons from Abeta-induced apoptosis. Furthermore, the present study indicates that inhibition of L-VSCC contributes to the neuroprotective effect of Gas6. PMID- 12527479 TI - Protective effects of estradiol against amyloid beta protein-induced inhibition of neuronal Cl(-)-ATPase activity. AB - Low concentrations of amyloid beta proteins (Abetas, 1-10 nM) were recently demonstrated to reduce Cl(-)-ATPase activity in parallel with an increase in the intracellular Cl(-) concentration ([Cl(-)]i) and decreases in plasma membrane phosphorylated phosphatidylinositol (PIP and PIP2) levels in cultured rat hippocampal neurons. In this study, 17 beta-estradiol (estradiol) at a therapeutic concentration (1.8 nM) for Alzheimer's disease was found to block these Abeta (Abeta25-35)-induced changes. This protective effect of estradiol on Cl(-)-ATPase activity was antagonized by a pure estrogen receptor antagonist, ICI182780 and inhibitors for cyclic GMP-dependent protein kinase (PKG) (KT5823), Ca(2+)-calmodulin-dependent protein kinase II (CaMKII) (KN62) and phosphatidylinositol (PI) 4-kinase (wortmannin and quercetin). Estradiol recovered Abeta-induced decreases in plasma membrane phosphoinositide (PIP and PIP2) levels, this effect being inhibited by KT5823 and KN62. Glutamate toxicity was augmented in neurons with elevated [Cl(-)]i either by Abeta-treatment or carbachol+KCl+LiCl-treatment. The increased glutamate toxicity in the Abeta treated neurons was attenuated by estradiol. Thus, a therapeutic concentration of estradiol protected Abeta-treated neurons against inhibition of Cl(-)-ATPase activity and an increase in [Cl(-)]i through its receptor, probably via PKG- and CaMKII(-)mediated recovery of PI4P formation. Elevated [Cl(-)]i may be related to enhancement of glutamate toxicity. PMID- 12527480 TI - Alpha 2C-adrenoceptor mediated regulation of cortical EEG arousal. AB - Alpha2-adrenergic drugs modulate cortical arousal and EEG. However, the role of individual alpha2-adrenoceptor (alpha(2)-AR) subtypes in these functions is not clear. We investigated the role of alpha(2C)-ARs in the modulation of baseline cortical EEG activity and EEG responses to the alpha(2)-AR selective agonist, dexmedetomidine (3-300 microg/kg, s.c.), and antagonist, atipamezole (3-1000 microg/kg, s.c.), by using alpha(2C)-AR knockout (KO) and wildtype (WT) mice. The overall amplitude (1-30 Hz) was not significantly altered in KO mice although the activity of theta band (4-8 Hz) was increased in these mice. The main finding was that dexmedetomidine (30-300 microg/kg) more effectively slowed and atipamezole (30-1000 microg/kg) less effectively increased cortical EEG arousal in KO mice compared to WT controls. Importantly, autoradiographical results showed no compensatory increase in other alpha(2)-AR subtypes in cortical, thalamic or other brain structures of KO mice. Furthermore, there were no differences between the genotypes in the levels of hippocampal choline acetyltransferase, monoamines or their metabolites. Altered baseline cortical EEG activity and EEG responses to alpha(2)-AR selective drugs in KO mice indicate that alpha(2C)-ARs are involved in regulation of cortical arousal. These results suggest that alpha(2C)-ARs may antagonize the sedative effect of alpha(2)-AR agonists mediated by activation of alpha(2A)-ARs. PMID- 12527481 TI - P2 receptors in the murine gastrointestinal tract. AB - The actions of adenosine, adenosine 5'-triphosphate (ATP), 2-methylthio adenosine diphosphate ADP (2-MeSADP), 2-methylthio ATP (2-MeSATP), alpha,beta-methylene ATP (alpha,beta-meATP) and uridine triphosphate (UTP) on isolated segments of mouse stomach (fundus), duodenum, ileum and colon were investigated. The localization of P2Y(1), P2Y(2), P2Y(4), P2X(1) and P2X(2) receptors and neuronal nitric oxide synthase (NOS) were examined immunohistochemically, and P2Y(1) mRNA was examined with in situ hybridization. The order of potency for relaxation of longitudinal muscle of all regions was: 2-MeSADP>/=2-MeSATP>alpha,beta meATP>ATP=UTP=adenosine. This is suggestive of P2Y(1)-mediated relaxation and perhaps a further P2Y receptor subtype sensitive to alpha,beta-meATP. As ATP and UTP are equipotent, the presence of a P2Y(2) receptor is indicated. ATP responses were inhibited by the P2Y(1)-selective antagonist MRS 2179, and suramin. P2Y(1) receptors were visualized immunohistochemically in the smooth muscle of the ileum and in a subpopulation for myenteric neurones, which also stained for NOS. P2Y(1) mRNA was localized in neurones in both myenteric and submucosal ganglia in the ileum. Taken together, these results suggest that ATP was acting on non adrenergic, non-cholinergic inhibitory neurons, which release both nitric oxide (NO) and ATP. Reduced relaxations to 2-MeSADP by tetrodotoxin and N(omega)-nitro L-arginine methyl ester, are consistent with this possibility. Adenosine acts via P1 receptors to relax smooth muscle of the mouse gut. Segments of mouse colon (in contrast to the stomach and small intestine) were contracted by nucleotides with the potency order: 2-MeSATP>alpha,betameATP>ATP; the contractions showed no desensitization and were antagonized by suramin and PPADS, consistent with responses mediated by P2X(2) receptors. Immunoreactivity to P2X(2) receptors was demonstrated on both longitudinal and circular muscle of the colon, but not in the other regions of the gut, except for a small subpopulation of myenteric neurones. In summary, neuronal P2Y(1) receptors appear to mediate relaxation, largely through NO in all regions of the mouse gut, and to a lesser extent by P2Y(1), P2Y(2) and a novel P2Y receptor subtype responsive to alpha,beta-meATP in smooth muscle, while P2X(2) receptors mediate contraction of colonic smooth muscle. PMID- 12527482 TI - Interleukin-2 regulates membrane potentials and calcium channels via mu opioid receptors in rat dorsal root ganglion neurons. AB - Our previous studies revealed that interleukin-2 (IL-2) exerted peripheral antinociception that was partially mediated by mu opioid receptors. No ionic explanations of this effect have yet been reported. The present study was designed to investigate effects of IL-2 on the physiological properties of capsaicin-sensitive small dorsal root ganglion (DRG) neurons, which are predominantly responsible for nociceptive transmission from the periphery to the spinal cord. Intracellualr recordings of DRG neurons were made in DRG/peripheral nerve preparation in vitro. IL-2 (10(3) U/ml) produced membrane hyperpolarization of -9.4 +/- 3.0 mV and this effect was blocked by beta-FNA (5 microM), a mu opioid receptor antagonist. Under whole-cell patch clamp recordings, transient high-threshold Ca(2+) currents were inhibited by -56.6 +/- 11.3% by IL-2. Simultaneous calcium imaging showed that this cytokine also inhibited depolarization-evoked increase in intracellular calcium concentration. All the effects of IL-2 were blocked by naloxone (1 microM). Consistent with previous studies, DAMGO, a selective mu opioid agonist, exerted similar inhibitory effects on membrane potentials and Ca(2+) currents. The present results indicated that mu opioid receptors were involved in the regulatory effects of IL-2 on membrane potentials and calcium channels in DRG neurons, which may contribute to IL-2 induced peripheral analgesia. PMID- 12527483 TI - Nitric oxide discriminates the sites and mechanisms of action of centrally acting anti-hypertensive drugs in rabbits. AB - The aim of the present study was to further investigate the mechanisms of the central hypotensive action of catecholamines and imidazolines, in particular the role of nitric oxide (NO). Microinjections into the nucleus reticularis lateralis (NRL/RVLM) located in the rostroventrolateral part of the medulla (RVLM) and/or into the nucleus tractus solitarii (NTS) were performed in pentobarbital anesthetized rabbits. Microinjections of brimonidine (1 ng/kg), which binds both alpha(2)-adrenergic receptors (alpha(2)-ARs) and I(1) imidazoline receptors (I(1)Rs), into the NRL/RVLM induced hypotension (69+/-2 vs. 88+/-2 mm Hg) (p<0.05). Microinjections of S23757 (1 microg/kg), an antagonist highly selective for I(1)Rs, into the same site, prevented the hypotensive effect of brimonidine. These data show that the hypotensive effects of low doses of brimonidine involve the I(1)Rs of the NRL/RVLM. Alpha-methylnoradrenaline (alpha-MNA) (0.5 microg/kg) microinjected into the NTS induced hypotension (76+/-4 vs. 91+/-4 mm Hg) (p<0.05). Microinjections of a low dose of brimonidine (1 ng/kg) into the NTS had no blood pressure (BP) effect at all. In contrast, a higher dose (10 ng/kg) acting on alpha(2)-ARs induced hypotension (72+/-3 vs. 96+/-2 mm Hg) (p<0.05). Nomega-Nitro-L-arginine (L-NNA) (1.5 microg/kg) injected into the NRL/RVLM prevented the hypotensive effect of both alpha-MNA and the higher dose of brimonidine injected into the NTS. Bicuculline (1.5 microg/kg) injected into the NRL/RVLM prevented the hypotensive effect of alpha-MNA injected into the NTS. It is demonstrated that (i) the activation of alpha(2)-ARs of NTS triggers a neuronal GABAergic pathway projecting to the NRL/RVLM region which is NO dependent (ii) both alpha(2)-adrenergic (NTS) and non-adrenergic I(1)R (NRL/RVLM) mechanisms account for the very powerful hypotensive effect of brimonidine, a compound with high affinities at both types of receptors. PMID- 12527484 TI - Behavioral action of ethanol in Porsolt's forced swim test: modulation by 3 alpha hydroxy-5 alpha-pregnan-20-one. AB - Ethanol is known to increase cortical and plasma content of GABAergic neurosteroid 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha,5alpha-THP) which is responsible for some of its behavioral and electrophysiological effects. We have previously demonstrated the antidepressant like effect of 3alpha,5alpha-THP in mice. This study investigated the role of 3alpha,5alpha-THP in acute, chronic and withdrawal effects of ethanol using mouse forced swim test (FST) paradigm. While acute systemic ethanol (2 or 2.5 g/kg) administration exhibited an antidepressant like effect, its prolonged consumption produced tolerance to this effect and its withdrawal, on the other hand, elicited enhanced behavioral despair (depression). The antidepressant like effect of ethanol was potentiated by GABA(A) receptor agonist, muscimol (0.5 mg/kg, i.p.), 3alpha,5alpha-THP (0.5, 1 or 2 microg/mouse, i.c.v.) and by neurosteroidogenic drugs viz. selective serotonin reuptake inhibitor (SSRI), fluoxetine (5 or 20 mg/kg, i.p.), agonist at mitochondrial diazepam binding inhibitor receptor, FGIN 1-27 (0.5 or 1 microg/mouse, i.c.v.), or 11beta-hydroxylase inhibitor, metyrapone (0.5 or 1 microg/mouse, i.c.v.) which are known to increase endogenous 3alpha,5alpha-THP content. Furthermore, inhibition of the endogenous neurosteroid biosynthesis by drugs like 5alpha reductase inhibitor, finasteride (50 mg/kg, s.c.), 3beta-hydroxysteroid dehydrogenase inhibitor, trilostane (30 mg/kg i.p.) or 3alpha-hydroxysteroid dehydrogenase inhibitor, indomethacin (5 mg/kg, i.p.) and GABA(A) receptor antagonist, bicuculline (1 mg/kg, i.p.) blocked the antidepressant like effect of ethanol. Withdrawal of ethanol from mice consuming it chronically displayed enhanced behavioral despair and elicited tolerance to antidepressant like action of acute ethanol (2.5, 3 or 3.5 g/kg). Moreover, sub-antidepressant doses (0.25 or 0.5 microg/mouse, i.c.v.) of 3alpha,5alpha-THP and fluoxetine (5 mg/kg, i.p.) but not imipramine (1 mg/kg, i.p.) reversed the depression associated with ethanol withdrawal indicating sensitization to their antidepressant action. Thus, 3alpha,5alpha-THP plays a pivotal role in the actions of ethanol and in the depression associated with ethanol withdrawal. These findings may be of potential ramification to contribute to the depression associated with alcoholism and its treatment using neurosteroids. PMID- 12527485 TI - Early gastric cancer: endoscopy, laparoscopy and surgery. PMID- 12527486 TI - Gastro-oesophageal reflux disease: a decade of changes. PMID- 12527487 TI - Subclavian artery aneurysms. AB - We report the management of 14 subclavian artery aneurysms (13 true, one false) occurring in seven male and seven female patients (average age, 48 years). The aetiology of the aneurysms included thoracic outlet syndrome in eight, atherosclerosis in five and infection in one patient. Twelve aneurysms were of extrathoracic location, while two aneurysms were intrathoracic. Symptoms related to subclavian artery aneurysms were present in 11 patients (compression in four, haemorrhage in one, and ischaemia in six patients), whereas three aneurysms were asymptomatic. All aneurysms were treated surgically. The supraclavicular approach was used in 11 cases, and the combined transsternal and supraclavicular approach was used in two cases. After aneurysm resection, the reconstruction was performed with end-to-end anastomosis in five cases and with saphenous vein or synthetic grafts in eight cases. One infected subclavian artery aneurysm was treated with carotid to axillary saphenous vein bypass after exclusion of the aneurysm. Five associated brachial embolectomies and one bypass from the axillary to the distal brachial artery were performed. In all thoracic outlet syndrome cases, decompression at the thoracic outlet was also performed. There was no operative mortality, and the early patency rate was 100%. The follow-up period was from 6 months to 10 years (mean, 3.92 years). During this period, one patient died of malignancy and one patient required reoperation due to aneurysmal degeneration of the saphenous vein graft. Surgical treatment is recommended for all patients with subclavian artery aneurysms to prevent potential complications. PMID- 12527488 TI - Clinical application of ultrasonic probing for preoperative staging of colorectal carcinoma. AB - OBJECTIVE: The aim of this study was to assess the value of ultrasonic probing (USP) on colonoscopy for the preoperative staging of colorectal carcinoma. METHODS: Seventy-five patients with colorectal carcinomas proven pathologically underwent USP (Olympus UM-2R, 12 MHz; UM-3R, 20 MHz) on colonoscopy before surgery for colorectal cancer. The results were compared with pathological findings of resected specimens. RESULTS: Colorectal carcinoma appeared as a hypoechoic mass on USP. USP had an overall accuracy rate of 82.7% for diagnostic T staging of colorectal carcinoma. In determining lymph node metastasis, the sensitivity and specificity were 53.2% and 61.5%, respectively. The positive and negative predictive values were 0.87 and 0.22, respectively. CONCLUSIONS: USP is valuable for the staging of colorectal carcinoma and has a high accuracy rate for determining the depth of tumour invasion. The preoperative information obtained by this tool may influence the choice of therapy. PMID- 12527489 TI - Endovascular repair for abdominal aortic aneurysms: initial experience of an endograft programme. AB - OBJECTIVE: To evaluate early clinical results of elective endovascular repair of abdominal aortic aneurysms during the initial phase of an aortic endograft programme and to compare them with conventional open surgery. METHODS: Between July 1999 and September 2001, all patients with infrarenal abdominal aortic aneurysms undergoing elective repair were studied. The results of endovascular repair were compared with those of conventional repair. RESULTS: Twenty-seven endovascular repairs (24 men and three women; mean age, 74 yr) and 25 conventional repairs (19 men and six women; mean age, 73 yr) for infrarenal abdominal aortic aneurysms were evaluated. The aneurysm diameters in the two groups were similar (mean, 6.1 cm in the endovascular repair group and 6.6 cm in the conventional repair group). The comorbidities of the two groups were also comparable. The duration of operation was longer in the endograft group (249 +/- 86 min vs. 206 +/- 56 min), while the blood loss was significantly less (600 +/- 486 mL vs. 1074 +/- 1220 mL). The length of stay in the Intensive Care Unit (ICU) and the overall duration of hospitalization was also significantly less in the endograft group (1 +/- 1 d vs. 3 +/- 2 d in ICU; 9 +/- 5 d vs. 13 +/- 6 d of hospitalization). There was one hospital death in each group (4%), and the complications were similar between the two groups. During a mean follow-up period of 11.6 +/- 7.5 months, there was no rupture or open conversion in the endograft group. CONCLUSIONS: In the initial phase of the aortic endograft programme, the mortality and morbidity were acceptable and comparable to that of open surgery. PMID- 12527490 TI - Surgical treatment and follow-up results of pituitary ACTH microadenoma: 18 years' experience. AB - OBJECTIVE: To review retrospectively our clinical experience with surgical treatment and follow-up of pituitary adrenocorticotropic hormone (ACTH) microadenomas at Rui Jin Hospital of Shanghai Second Medical University. METHODS: Eighty-seven patients with ACTH microadenomas underwent surgery via the transsphenoidal approach. Preoperative computed tomography and/or magnetic resonance imaging revealed microadenomas in only 46 patients and was negative in the remaining 41 patients, which were classified as microadenoma grade 0. High dose dexamethasone (Dx) suppression testing was positive in 86% of patients and low-dose Dx suppression testing was negative in all patients. RESULTS: Histological confirmation of ACTH microadenoma was made in all 87 cases, with a minimum tumour size of only 1 to 2 mm in diameter. The remission rate was 82% for grade I and 92% for grade 0 microadenomas during 2 to 18 years of follow-up. CONCLUSIONS: Neuroimaging studies have limited value in diagnosing ACTH grade 0 microadenoma. Clinical manifestation combined with positive Dx suppression testing is decisive in these circumstances. Surgical intervention could achieve a high relief rate for ACTH microadenomas, especially in grade 0 group. PMID- 12527491 TI - Double-blind, randomized, placebo-controlled trial to determine the efficacy of eutectic lidocaine/prilocaine (EMLA) cream for decreasing pain during local anaesthetic infiltration for out-patient haemorrhoidectomy. AB - OBJECTIVE: The study was undertaken to evaluate the efficacy of eutectic lidocaine/prilocaine (EMLA) cream for decreasing pain during local anaesthetic infiltration for outpatient haemorrhoidectomy. METHODS: Ninety-eight patients were randomly assigned either to receive EMLA or placebo. The creams were applied 45 minutes prior to injection of a lidocaine/bupivacaine mixture using a diamond shaped perianal block. All participants were blinded to the specific medication received. They were asked to rate pain and levels of acceptability using a pre validated pain scale and questionnaire. RESULTS: There were 49 patients in each group. The baseline characteristics between the two groups were similar. Forty patients (82%) in the EMLA group and 42 patients (86%) in the placebo group reported only mild pain during injection and infiltration of the lidocaine/bupivacaine mixture. The mean rank pain scores were 49.11 and 48.89, respectively (p = 0.886, not significant). CONCLUSION: While outpatient haemorrhoidectomy under local anaesthesia was generally well tolerated, there was no statistically significant difference between EMLA cream and placebo for decreasing pain during anaesthetic infiltration. PMID- 12527492 TI - P53 protein expression in transitional cell carcinoma of the bladder - experience of the University of Malaya Medical Centre. AB - OBJECTIVES: To study the incidence of p53 oncoprotein overexpression and its relationship to tumour grade, stage and clinical prognosis in a cohort of local Malaysian patients. METHODS: All cases of transitional cell carcinoma (TCC) of the bladder diagnosed and treated at the University of Malaya Medical Centre from January 1995 to December 2000 were retrieved from the hospital records. Sections from paraffin-embedded tissues were retrieved and stained for p53 oncoprotein using immunohistochemistry techniques. P53 oncoprotein results were analyzed in relation to tumour grade, stage and clinical prognosis. Fisher's exact test was used to evaluate the relationship between categorical variables and the Kaplan Meier procedure was used to assess survival outcomes. The Cox regression model was used for multivariate analysis. RESULTS: A total of 64 cases were studied. The mean follow-up period was 23.7 months. The number of p53 positive cases was significantly higher in high-grade (G3) (p = 0.006) and muscle-invasive tumours ( summation operator T2, p = 0.035). The status of p53 expression had no significant association with recurrence-free (p = 0.594) or overall survival (p = 0.955). In multivariate analysis, a multiplicity of tumours at presentation (p = 0.004) and a history of cigarette smoking (p = 0.016) were independent predictors of recurrence. Tumour stage (p = 0.024) was the single independent predictor for poor overall survival. CONCLUSIONS: Overexpression of p53 is associated with TCC of higher grade and tumour stage. It had no significant impact on prognosis in this cohort of TCC cases. PMID- 12527493 TI - Atypical ductal hyperplasia of the breast in young women: two case reports. AB - Atypical ductal hyperplasia of the breast is a benign proliferative condition that is associated with an increased risk of development of breast cancer in either the ipsilateral or contralateral breast. Following diagnosis at biopsy, respective management options range from observation to chemoprophylaxis to prophylactic surgery. We present two cases in young women, facing prolonged follow-up, one managed with observation only, and the other managed with ipsilateral mastectomy and reconstruction. PMID- 12527494 TI - Large adrenocortical carcinoma extending into the inferior vena cava and right atrium. AB - A case of large adrenocortical carcinoma extending into the inferior vena cava and right atrium is reported. Computed tomography showed a large mass displacing the left kidney inferiorly with an intravascular tumour thrombus extending into the inferior vena cava and right atrium. Radical surgery under hypothermia and cardiopulmonary bypass was performed and the tumour mass, together with the tumour thrombus, was successfully removed. The presence of intravascular tumour extension alone should not be a contraindication to radical surgical therapy, as it is the best hope for prolonged survival. PMID- 12527495 TI - Laparoscopic pancreatic resection of an insulinoma in a child. AB - Insulinoma is a rare neuroendocrine tumour infrequently reported in children. Operative excision is the treatment of choice, but requires a disproportionately large incision to remove a small and benign tumour located deep in the retroperitoneum. Laparoscopic resection of benign tumours of the pancreas has been increasingly reported in adults and seems to be ideal for pancreatic insulinomas. We present the case of pancreatic insulinoma in a 13-year-old boy treated by laparoscopic distal pancreatectomy. PMID- 12527496 TI - Synchronous triple early cancers occurring in the stomach, colon and gallbladder. AB - We report an unusual case of synchronous triple early cancers of the stomach, gallbladder and sigmoid colon. The patient was a 70-year-old man. Gastrointestinal endoscopy at a medical check-up initially revealed early gastric cancer. A thorough preoperative investigation showed a gallbladder tumour and early colon cancer. Endoscopic mucosal resection was performed for the colon cancer. Microscopically, the specimen showed well-differentiated adenocarcinoma involving the submucosal layer. Distal partial gastrectomy (D1) and simple cholecystectomy with lymph node dissection were also performed. The histopathological diagnosis was well-differentiated adenocarcinoma in the stomach (T1, N0, M0; Stage IA) and poorly differentiated papillary adenocarcinoma (T1, N0, M0; Stage I) in the gallbladder. The patient was doing well with no evidence of recurrence 6 months after surgery. PMID- 12527497 TI - Portal vein thrombosis after radiofrequency ablation for recurrent hepatocellular carcinoma. AB - Recurrent hepatocellular carcinoma (HCC) deserves multidisciplinary treatment in addition to surgical resection. Radiofrequency ablation (RFA) is an evolving, localized, thermal ablative treatment for unresectable hepatocellular carcinoma (HCC). Though the preliminary results of RFA in clinical studies are encouraging, its serious complications should not be underestimated. Portal vein thrombosis as a result of direct blood vessel injury by RFA is rarely reported and is potentially fatal in patients with limited liver reserve due to underlying liver cirrhosis. We present a case of portal vein thrombosis as a complication of RFA treatment for recurrent HCC and illustrate its underlying possible mechanism. PMID- 12527498 TI - Mesenteric ischaemia. PMID- 12527499 TI - Enteric dysfunction in patients with asymptomatic HIV-infection versus patients with hypoproteinemia/hypalbuminemia and advanced HIV-infection. AB - BACKGROUND: It is unclear whether enteric dysfunction and protein losses contribute to hypoproteinemia, which heralds poor survival in HIV infection. METHODS: We investigated alpha-1-antitrypsin-clearance (AAT-CL), D-xylose resorption and total gut transition time in 14 HIV+ patients with hypoproteinemia (serum protein < 6 g/dl, albumin < 3 g/dl, median CD4-cell count 58/microl; (group I)), in 10 asymptomatic HIV+ patients (median CD4-cell count 290/microl, (group II)) and in 15 healthy volunteers (group III). RESULTS: AAT-CL in group I (16.5 (2.9 278.2) ml/d; median (range)) was higher than in groups II (9.5 (1.7 23.1) ml/d) and III (10.6 (0.8 19.5) ml/d; p = 0.0114). Likewise, D-xylose recovery was on average threefold lower in group I than in groups II (p = 0.0009) and III ( p < 0.0001), whereas total gut transition time was significantly shorter in both HIV-infected groups (group I: 49.8 h (23.5-72.7), p=0.0431; group II: 32.6 h (23-54.6), p=0.0104) than in the healthy controls (group III 61.6 h (39.1-87.7)). CONCLUSIONS: Thus, impaired intestinal resorption and enteral protein losses may contribute to hypoproteinemia in advanced HIV infection, whereas accelerated intestinal motility may be present already in asymptomatic stages of HIV infection. PMID- 12527500 TI - IgM-enriched immunoglobulin preparation for immunoprophylaxis in cardiac surgery. AB - OBJECTIVE: Evaluating the effects of prophylactic administration of IgM-enriched immunoglobulins (IVIG) on immunological- and clinical parameters in cardiac surgical patients. PATIENTS AND METHODS: 41 patients were randomized to receive either an IgM-enriched immunoglobulin (Pentaglobin(R)) preparation (1,300 ml immunoglobulin, equivalent to 65 g protein) combined with routine antibiotic prophylaxis (Group A; n = 20, 1 drop-out), or routine antibiotic prophylaxis plus placebo (Group B; n = 20). Patients were comparable with respect to their APACHE II score, comorbidity, coronary risk, operating time, clamp, and ischemic time. Endotoxin and endotoxin neutralizing capacity (ENC) were determined by a kinetic turbidimetric Limulus amebocyte lysate (LAL) assay with internal standardization. Serum levels of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF)-alpha, soluble TNF-Receptor I (sTNF-R1), and interleukin-10 (IL-10) were determined by ELISA. Data analysis was performed by area under the curve (AUC) calculation and ANOVA for endotoxin neutralizing capacity and by ANOVA for all other cases. RESULTS: All patients survived. Endotoxin plasma levels were generally but not significantly higher in group A than in controls, while the difference in endotoxin neutralizing capacity (ENC) reached significance. IL-6, TNF-alpha, IL 10 and TNF-R1 were not different between both groups, however. There were significantly less patients with signs of inflammation (fever, leukocytosis, hypotension) in group A (group A n = 2; group B n = 9; p<0.05). This was paralleled by a slightly reduced hospitalization period in group A patients compared to group B patients (A:12.05 +/- 3.66 vs. B:13.45 +/- 3.72 days; n.s.). All data are given as mean +/- standard deviation (SD). CONCLUSION: The results of this study support that IgM-enriched IVIG preparation are effective when used prophylactically in patients undergoing procedures with cardiopulmonary bypass. The mechanisms of endotoxin neutralization and the effect of the host immune status on the efficacy of IVIG treatment remain to be elucidated. PMID- 12527501 TI - Dental health of elderly in retirement homes of two cities in south Croatia--a cross-sectional study. AB - OBJECTIVE: A survey of dental health status was conducted in institutionalized elderly in South Croatia in order to assess the oral health of the elderly population. PARTICIPANTS: In 274 institutionalised elderly from five retirement homes in Split and Korcula we registered dental status according to World Health Organization criteria. The mean age of the patients was 81 +/- 8; there were 220 female and 54 male subjects. RESULTS: A total of 192 (70.0%) were totally edentulous, and additional 39 (14.2%) were edentulous in one jaw. A significantly higher prevalence of edentulousness with the increasing age was recorded. The mean number of remaining sound teeth, decayed teeth and radices relictae in the elderly decreased with increasing age. CONCLUSIONS: The elderly population in retirement homes in South Croatia has poor dental health. This calls for action to improve dental health and implement some special features in the dental health service programs for the institutionalised elderly. PMID- 12527502 TI - Radioimmunotherapy with 153Sm-CEA monoclonal antibody in nude mice bearing human colon carcinoma: an experimental study. AB - OBJECTIVE: To observe the therapeutic effect of 153Sm-labeled CEA monodonal antibody(mAb) in nude mice bearing human colon carcinoma. METHODS: Fifteen nude mice were subjected to subcutaneous inoculation of human colon carcinoma cells, and 3 days later, they were divided into 3 groups with equal numbers to receive single high-dose injection of 11.1 MBq 153Sm-CEA mAb (therapy group), 11.1MBq 153SmCl3 (therapy control group), or 100 microl normal saline (non-treatment control group). The tumor-inhibiting effect of 153Sm-labeled CEA mAb was evaluated in terms of body weight changes and tumor volume variation 4 weeks after the treatment. Histological analysis of tumors were performed in all the groups after the all other observations were completed. RESULTS: 153Sm-CEA mAb had a significant anti-tumor effect, with a tumor inhibition rate of 74.29% at 4 weeks after treatment, while for 153SmCl3, the inhibition rate was only 15.90%. Rapid tumor growth was observed in non-treatment control group. No significant difference in the body weight changes was noted between the 3 groups. Histopathological examination revealed tumor necrosis as the evidence for radioactive damage in therapy group, which was not observed in non-treatment control group. CONCLUSION: 153Sm-CEA mAb has a strong selective inhibitory effect against colon carcinoma and may be potentially used as an agent in radioimmunotherapy. PMID- 12527503 TI - [Role of phorbol 12-myristate 13-acetate on cytokine expression in JAR trophoblast cell line]. AB - OBJECTIVE: To investigate whether phorbol 12-myristate 13-acetate (PMA) modifies the invasive ability of trophoblast cells by regulating their cytokine productions. METHODS: Reverse transcriptase-polymerase chain reaction was used to examine the effect of PMA on the expression of cytokines which regulated the invasive ability of trophoblast cells. RESULTS: Prior to PMA treatment, expressions of the cytokins including hepatocyte growth factor (HGF), interleukin (IL)-1beta, insulin-like growth factor (IGF)-II, transforming growth factor (TGF) beta and vascular endothelial growth factor (VEGF) were all detected in JAR cells, only with the exception of IGF-I. After incubation with 100 nmol/L PMA for 24 h, the cells showed strong expression of IL-1beta, HGF and IGF-II, with reduced expression of TGF-beta2 and TGF-beta3. CONCLUSION: By regulating the autocrine of these cytokines, PMA exercises its effect to enhance the invasive ability of trophoblast or choriocarcinoma cells. PMID- 12527504 TI - [Effects of advanced glycation end-products on cell cycle distribution and apoptosis in neonatal rat cardiac myocytes]. AB - OBJECTIVE: To investigate the effects of advanced glycation end-products (AGEs) on cell cycle distribution and apoptosis in neonatal rat cardiac myocytes (CMs) cultured in vitro. METHODS: CMs already cultured for 3 to 5 d in vitro were continuously cultured with DMEM supplemented with 1% or 10% fetal bovine serum for 24 h, prior to exposure to AGE-modified human serum albumin (AGE-HSA, at 50 and 100 microg/ml) for 12, 24 and 48 h. Cells cultured in the same manner but without AGE-HSA treatment served as the control group. All cells were collected and analyzed by flow cytometry for cell cycle distribution and cell apoptosis, and the number of apoptotic body/nucleus of CMs was determined by in situ cell death detection kit (fluorescein). RESULTS: AGEs had no obvious effects on cell cycle distribution in CMs, but could increase the number of apoptotic body/nucleus of CMs in a time-dependent manner to up to 0.55 and 1.23 times at 24 and 48 h respectively, as compared with the number at 12 h in control cells. AGEs at 50 and 100 microg/ml increased the number of apoptotic body/nucleus of CMs to various degrees at different time points: 0.74 and 1.21 times respectively at 12 h; 1.15 and 1.78 times at 24 h; 0.83 and 1.19 times at 48 h. The average number of apoptotic body/nucleus of the cells treated with 100 microg/ml AGEs at 12, 24, 48 h were 0.27, 0.29, 0.20 times respectively that of the cells with 50 microg/ml AGEs treatment. CONCLUSIONS: AGEs do not affect the distribution of cell cycle but can impair neonatal rat CMs by increasing their apoptosis rate. PMID- 12527505 TI - [Screening of short peptides that bind specifically to osteosarcoma cells by phage-displayed peptide library]. AB - OBJECTIVE: To obtain short peptides which bind specifically to osteosarcoma cells os-732 by means of screening from 12 peptide libraries. METHODS: Osteosarcoma cells os-732 were used as the target cells and osteoblasts as the absorber cells for subtraction biopanning from a 12-mer peptide phage-display library. After 3 rounds of screening, the positive phage clones were identified by cell enzyme linked immunosorbent assay (ELISA) and immunohistochemistry, and the amino acid sequences were deduced by DNA sequencing. RESULTS: Nine positive phage clones screened out of 20 clones showed specific binding with osteosarcoma os-732, but no conserved motif was found in these peptides. CONCLUSION: The specific peptides screened from the phage library may be used as potential candidates as ligands for tumor-targeting therapy. The results also suggested that there are different epitopes on the surface of tumor cells. PMID- 12527506 TI - [HBsAg uptake into human trophoblasts cultured in vitro]. AB - OBJECTIVE: To detect the entry of HBsAg into human trophoblasts cultured in vitro, to provide some clues for the mechanism responsible for HBV intrauterine transmission. METHODS: Digestion with both trypsin (0.25%) and Dnase I(0.15 U/ml) followed by repeated purification was performed to obtain a homogeneous population of trophoblast cells. The well-developed cells during longitudinal phase were randomly divided in 6 groups, which were co-incubated respectively with HBsAg-anti-HBs complex (I), co-culture of heat-inactivated sera positive for HBsAg, HBeAg, and anti-HBc and sera with high titer anti-HBs(II), heat inactivated sera positive for HBsAg, HBeAg, and anti-HBc(III), HBsAg (IV), heat inactivated normal sera (V), and normal medium(VI). The coverglasses mounted with confluent cells were harvested for anti-HBs immunohistochemical staining. RESULTS: The isolated cells comprised a highly purified villous trophoblast population and could meet the demands of further experiments. After co-incubation of the cells with different agents, HBsAg test yielded positive results in the cells co-incubated with the HBsAg-anti-HBs complex and in the co-cultures of heat inactivated sera positive for HBsAg, HBeAg and anti-HBc with sera containing high titer anti-HBs. HBsAg test was negative in all other cell groups. CONCLUSION: Human trophoblast cells could take in HBsAg in the form of HBsAg-anti-HBs complex instead of single HBsAg molecule. PMID- 12527507 TI - [Effect of hyperthermia on tight junctions between endothelial cells of the blood brain barrier model in vitro]. AB - OBJECTIVE: To investigate the effect of hyperthermia in vitro on tight junctions between the endothelial cells of the blood-brain barrier. METHODS: An in vitro blood-brain barrier model was established by coculture of ECV304 cells with astrocytes. Transendothelial resistance (TER) of in vitro blood-brain barrier was determined by Millicell-ERS system, and the morphological change of tight junctions examined by silver staining. The expression level of zonula occluden 1(ZO-1) was analyzed by semi-quantitative reverse transcriptase-PCR. RESULTS: The integrity of tight junctions was disrupted and the expression level of ZO-1 decreased after treatment with hyperthermia. CONCLUSIONS: Tight junctions between the endothelial cells of the blood-brain barrier can be destroyed by hyperthermia, and decreased expression level of ZO-1 induced by hyperthermia is one of the most important molecular mechanisms. PMID- 12527508 TI - [Cloning and sequence analysis of tomato fruit-specific E8 promoter from Lycopersicon esculentum (Zhongshu No.5)]. AB - OBJECTIVE: To obtain the gene encoding tomato fruit-specific E8 promoter therefore to prepare for exogenous gene transcription and expression in transgenic tomato fruit. METHODS: The cotyledons of tomato Lycopersicon esculentum (Zhongshu No.5) were collected for extracting the genomic DNA of this plant. The fruit-specific E81.1 and E82.2 promoter DNA were then amplified by PCR, the product of which was subcloned into pGEM-T vector. After identification by restriction enzymes, the recombinant T-vectors were subjected to sequence analysis. RESULTS: The fragments of the promoter as amplified by PCR were of predicted length. Digestion with Xba I and Hind III /BamH I proved correct insertion of the target fragments with expected length into the recombinant T vectors. As indicated by homology analysis, the resultant tomato fruit-specific E8 promoter was highly conservative, and E82.2 promoter of Zhongshu No.5, with GenBank submission number of AF515784, proved to share 99% homology with E82.2 promoter of Zhongshu No.5 Cherry as reported by Deikman J. CONCLUSION: Tomato fruit-specific E8 promoter of Zhongshu No.5 has been successfully cloned, thus making possible the subsequent research in oral vaccine of transgenic tomato. PMID- 12527509 TI - [Irradiated porcine skin impregnated with fluorine and silver covered by autogenous granule skin graft for treating severe burn injury]. AB - OBJECTIVE: To study the feasibility of using irradiated porcine skin impregnated with fluorine and silver covered by autogenous skin granule graft in the treatment of severe burn injury. METHODS: Autogenous split-thick skin grafts were cut into small pieces (<1 mm3), which were transferred onto the sheet of porcine skin mounted with a layer of Xinnaxing ointment (2 mm in thickness). After eschar removal, the burn wound was covered with the prepared porcine skin, sutured and fixed with small holes at the lower site for drainage. The growth of autogenous granule skin was then observed and the criteria for therapeutic effect evaluation drafted. RESULTS: The layer of Xinnaxing ointment on the the porcine skin improved the adhesion of the autogenous small skin grafts, therefore full use could be made of the donor skin source. In the 15 patients receiving this treatment, the graft-covered area was (12.3+/-3.5) % that of the wound, with the donor-recipient area ratio of 1: (11.0+/-2.7) and subsequent healing time of 27.5+/-5.5 d. Primary healing was achieved in 9 cases, and secondary healing in 6, demonstrating similar clinical effect to that of allo-skin grafting. CONCLUSION: The method is easy and economic, which saves autogenous skin source and produces satisfactory clinical effect. PMID- 12527510 TI - [Inhibitory effect of recombinant human endostatin on angiogenesis and lung metastasis of mouse lung adenocarcinoma LA795]. AB - OBJECTIVE: To study the inhibitory effect of recombinant human endostatin (rhES) on the angiogenesis and lung metastasis of mouse lung adenocarcinoma LA795. METHODS: The recombinant yeast strain containing the gene sequence encoding highly soluble rhES was induced by methanol for rhES production, which was purified with heparin affinity chromatography. T739 mice with subcutaneous inoculation of LA795 cells were randomized into 2 groups (10 in each group) to receive injection of either rhES (20 mg/kg x b x w x per day) or PBS in the same volume for 14 consecutive days starting from the sixth day after the inoculation. The angiogenesis and lung metastasis of the implanted tumors were subsequently observed. RESULTS: Purified rhES was successfully obtained. As shown by immunohistochemistry, the tumors in the mice receiving rhES treatment exhibited less density of the microvessels than those in the PBS-treated mice did (P<0.01). Pathological examination of the lung tissue of the mice in rhES group found no visible signs of tumor metastasis, which, in contrast, was widespread in PBS group. The weight of the lungs was also significantly different (P<0.01). CONCLUSION: rhES possesses good biological properties and can potently inhibit the angiogenesis and lung metastasis of mouse lung adenocarcinoma LA795. PMID- 12527511 TI - [Biological analysis of a novel gene related with interleukin-6]. AB - OBJECTIVE: To understand the action mechanism of interleukin (IL)-6 through investigation of its related genes. METHODS: Using electronic cloning with IL-6 related expressed sequence tags (ESTs), a novel gene at its full length of 924 bp was acquired. Reverse transcriptase-PCR method was subsequently employed to amplify the cDNA of this gene from the total RNA of human U937 cells, which were previously activated with 100 ng/ml IL-6 for 8 h. The fragment was then ligated into the pGEM-Teasy vector and sequence analysis was conducted. RESULT: The whole cDNA gene was successfully fished. CONCLUSION: Electronic cloning technology can be instrumental in identifying novel genes. PMID- 12527512 TI - [Preparation and identification of monoclonal antibodies against Penicillium marneffei]. AB - OBJECTIVE: To prepare and identify monoclonal antibodies (mAbs) against Penicillium marneffei. METHODS: Recombinant mannoprotein1 (MP1) of Penicillium marneffei was used to immunize BALB/c mice, and anti-MP1 mAbs were obtained by means of hybridoma. Screening and identification of the mAbs were subsequently performed with indirect enzyme-linked immunosorbent assay and Western blotting, respectively. RESULTS: Four hybridomas producing antibodies against Penicillium marneffei were obtained, and the IgG isotypes of the 4 mAbs were identified as IgG1 with affinity constants (K) of 8.2x10(-9), 4.7x10(-9), 6.5x10(-9) and 2.7x10(-9), respectively. Western blotting demonstrated specific recognition of Aspergillus fumigatus MP1 by the obtained mAbs. CONCLUSION: The 4 hybridomas producing anti-MP1 mAbs with high specificity and affinity can be of significant value in the diagnosis of Penicilliosis marneffei infections. PMID- 12527513 TI - [Morphological observation of mesenchymal stem cells cultured with allogenic decalcified bone matrix]. AB - OBJECTIVE: To investigate the feasibility of using decalcified bone matrix loaded with mesenchymal stem cells (MSCs) derived from adult human bone marrow as scaffolds for bone tissue engineering. METHODS: Allogenic decalcified bone matrix was prepared. MSCs were isolated from adult human bone marrow with monocyte separating medium and then cultured in basic medium of mesenchymal stem cells (MSC). The third generation of the cultured cells were identified by FITC antiCD105, seeded onto allogenic decalcified bone matrix followed by observation under scanning electron microscope and invert microscope a week later. RESULTS: Rapid proliferation of both primary and subcultured MSCs was observed, and CD105 positive cells reached 64.1% among the third generation of the cells. MSCs also grew well and proliferated rapidly after being cultured with decalcified bone matrix. CONCLUSION: Naive MSCs are ideal seeding cells for bone tissue engineering, and the allogenic decalcified bone matrix loaded with MSCs may be used as a good scaffold. PMID- 12527514 TI - [Pharmacokinetics of Nolaterex in tumor cells]. AB - OBJECTIVE: To investigate the relationship of the transportation characteristics of Nolaterex, a new anti-cancer drug, with its sensitivity in tumor cells. METHODS: The sensitivities of 3 tumor cell lines (C6, SRS-82 and LoVo) to nolaterex were determined by growth inhibition test. After Nolaterex exposure, the intracellular drug concentration was measured by high-performance liquid chromatography. RESULTS: C6 was the most sensitive among the 3 cell lines, and the IC50 values of SRS-82 and LoVo cells were 6.8-fold and 13.8-fold higher, respectively, than that of C6 cells. In all the 3 cell lines, linear relationship between intracellular and extracellular drug concentrations was noted. The intracellular steady-state level achieved in C6 was significantly higher than the levels in the other two cell lines. CONCLUSIONS: The results indicate that nolaterex quickly enters the cells, and different cell lines may have different nolaterex-transporting capacities, thus partially accounting for different states of sensitivity of the tumor cells to nolaterex. PMID- 12527515 TI - [Therapeutic effect of L-arginine on traumatic shock in rats]. AB - OBJECTIVE: To evaluate the therapeutic effect of L-arginine (L-Arg) on traumatic shock in rats and explore the possible mechanisms. METHODS: Rat models of traumatic shock were established in Sprague-Daulay rats, which were randomly divided into two groups either to receive L-Arg treatment or not. The plasma concentration of endothelin (ET) and oxygen partial pressure in the tissues from the skeletal muscles, liver and small intestine were measured before and after the shock and 1, 3, and 5 h after resuscitation. The hemodynamics of the rats and their survival rates at 12 and 24 h were recorded. RESULTS: The changes of plasma ET levels and oxygen partial pressure in tissues of both groups were statistically significant after traumatic shock (P<0.05). Plasma ET concentration at 5 h after resuscitation was significantly lower in the treatment group than in the non-treatment shock group (P<0.05), while oxygen partial pressure in the liver and small intestine after resuscitation were significantly higher in the treatment group (P<0.05). The survival rates at 12 and 24 h were also significantly different between the 2 groups. CONCLUSION: Adscititious L-Arg can decrease plasma ET levels, improve oxygen partial pressure of internal organs and significantly increase the survival rate of the rats with traumatic shock. PMID- 12527516 TI - [Construction of online medical parasites database]. AB - We aim to construct a online medical parasites database from which information can be retrieved through key words, ASP program and SQL language. The connection between the web pages and the database was established to enable relevant web page searching and parasitological information retrieval on the basis of the database files. A method for online searching through the database is elaborated in this paper by introducing the construction of the medical parasites database. PMID- 12527517 TI - [A method for transfering physiological data through GSM wireless communication network]. AB - This article describes a method for transfering physiological data through GSM wireless communication network. The data of heart rate and body temperature are collected by a microcontroller and transmitted through serial ports to the GSM data module, which then transmits the data to a remote terminal through GSM network. This method ensures cheap and stable long-distance data transfer within the coverage of the GSM network. PMID- 12527518 TI - [Value of conformal radio therapy for middle stage or advanced primary hepatocellular carcinoma]. AB - OBJECTIVE: To evaluate the value of conformal radio therapy in the treatment of primary hepatocellular carcinoma (HCC) in advanced stages. METHODS: Altogether 198 patients with advanced HCC underwent three-dimensional conformal radio therapy (3D-CRT) in a dose range 48 to 63 Gy in 6 to 9 fractions completed in a course of 12 to 18 d. The total clinical tumor volume (CTV) received more than 90% of the prescribed irradiation dose delivered through 6MV X ray. CT examinations were performed before 3D-CRT and in the three-year follow-up period to verify the therapeutic effects. RESULTS: Partial tumor remission was achieved in 81 cases (41%) 1 month after the treatment and 95 cases (48%) 3 months later. Ten cases (5%) failed to respond to the treatment, while the majority (186, 94%) showed tumor remission 6 months after the treatment, with only 2 cases (1%) had progression of the tumors. In the 48 patients with cancer embolus in the portal vein, 31 (65.6%) showed evidence of diminished embolus. The alpha-fetoprotein levels in the HCC patients were reduced significantly after the treatment (P=0.015). Among the stage-III patients 1-year survival rate was 68%, 2-year survival rate was 41%, and 3-year survival rate was 35%. CONCLUSIONS: 3D-CRT is an effective method for treating advanced HCC. PMID- 12527519 TI - [HLA genotyping by automatic semi-quantitative polymerase chain reaction-reverse sequence specific oligonucleotide]. AB - OBJECTIVE: To explore a HLA genotyping method that can be used for organ transplantation tissue typing, and especially for establishing hemopoietic stem cell bank and the cord blood stem cell bank by analyzing the PCR-DNA. METHODS: Modified automatic method based on semi-quantitative amplification system of HLA I, I allele genotyping was established using reverse sequence-specific oligonucleotide with polymerase chain reaction (PCR-RSSO), and was compared with PCR (using sequence specific primer, PCR-SSR) and manual PCR-RSSO in terms of the accuracy, resolution, and the quantity of DNA consumption. A total of 635 blood samples were genotyped with HLA-A, B, C, DR and DQ alleles using auto semi quantitative PCR-RSSO, 166 of which were also examined using PCR-SSP and manual PCR-RSSO simultaneously. RDSULTS: The success rate of automatic semi-quantitative PCR-RSSO, PCR-SSP and manual PCR-RSSO was 98.4% (3 124/3 175), 98.8% (656/664) and 88.3% (732/830) respectively, with no significant difference between the former 2 as indicated by X2 test (P>0.05). Auto semi-quantitative PCR-RSSO, however, yielded significantly higher success rate than manual PCR-RSSO (P<0.05). CONCLUSION: PCR-RSSO is capable of identifying 706 alleles of HLA-I, II antigens which amounts to 75.43% of the 936 alleles published by WHO in 2000, with intermediate to high resolution, even in the case of the homozygote. The hybridization results documenting the original data can be conveniently preserved. This method therefore possesses the merits of low expenses, low labor intensity, and rapid processing of large number of samples. PMID- 12527520 TI - [Magnetic resonance imaging enhanced by superparamagnetic iron oxide particles in detecting lymph node metastasis of tumors: an experimental study]. AB - OBJECTIVE: To investigate the value of superparamagnetic iron oxide particles (SPIO) in magnetic resonance (MR) imaging for detecting lymph node metastasis of tumors. METHODS: Twelve New Zealand rabbits were used in this study, 6 of which received unilateral intra-muscular inoculation of VX2 carcinoma cells to induce lymph node metastasis of the tumor, and the other 6 served as normal control group. MR images of the lymph nodes of the rabbits were obtained before and 12 h after subcutaneous injection of SPIO, followed by image analysis in correlation with pathological examinations. RESULTS: On plain MR images, normal and metastatic lymph nodes showed similar signal characteristics. After administration of SPIO, the signal intensity of both normal lymph nodes and metastatic ones remained unchanged in spin echo (SE) T1-weighted images. On SE T2 weighted images, the signal intensity of normal lymph nodes significantly decreased heterogeneously, while that of all metastatic lymph nodes remained unchanged. In gradient recalled echo (GRE) T2-weighted images, the signal intensity of normal lymph nodes decreased significantly and homogeneously, while that of 4 rabbits in metastasis group remained unchanged, with the signal intensity in the other 2 rabbits decreased heterogeneously. CONCLUSION: SPIO enhanced MR imaging can be applied to detect lymph node metastasis of the tumors. PMID- 12527521 TI - [Protective effects of preconditioning phlebotomy against myocardial ischemia reperfusion injury in rat]. AB - OBJECTIVE: To investigate the protective effect of preconditioning phlebotomy against ischemia-reperfusion injury and the role of calcitonin gene-related peptide (CGRP) in this protection mechanism. METHODS: Sixty SD rats were randomly assigned in equal number into control, phlebotomy, and CGRP polyclonal antibody (PcAb) groups, all of which were subjected to 30 min regional ischemic followed by two-hour reperfusion, initiated 15 min after phlebotomy in the latter 2 groups. The rats in CGRP PcAb group were given the antibody (2.5 mg/kg) before phlebotomy. After the operations, plasma CGRP level was determined in 8 rats from each group by radioimmunoassay, and the infarct size (IS) by nitro blue tetrazolium, represented as percentage of the area at risk (AAR). RESULTS: The mean plasma CGRP level was significantly increased in phlebotomy group compared with the levels in control and PcAb groups (F=12.4, P=0.032-0.005), and a significant reduction in IS (F=52.4, P<0.001) and IS/AAR (F=43.7, P<0.001) occurred in phlebotomy group. CONCLUSION: Preconditioning phlebotomy has obvious cardioprotective effect against ischemia-reperfusion injury, in the mechanism of which CGRP may play a role. PMID- 12527522 TI - [Preparation of famotidine microemulsion and its quality evaluation]. AB - OBJECTIVE: To study the preparation of famotidine microemulsion (FM) and its quality evaluation, thereby to establish the quality-control criteria for this preparation. METHODS: The formulations of the FM were optimized by studying the pseudoternary phase diagrams and its absorption in rat jejunum using in situ perfusion method. The morphology, particle size distribution, famotidine concentration and stability of the FM were studied. RESULTS: The optimization of formulation was completed successfully. Electron microscopy presented the FM as small spherical drops, with a mean diameter of 65 nm. The average recovery as determined by high-performance liquid chromatography was 97.63%, its mean RSD being 0.72%. CONCLUSION: The FM is easy to prepare with consistent quality, whose reliable determination method may be easily reproduced. PMID- 12527523 TI - Clinical significance of positron emission tomography for lymphoma patients. AB - OBJECTIVE: To explore the characteristics and clinical significance of positron emission tomography (PET) examination for malignant lymphoma. METHODS: PET examination was performed in 20 patients with malignant lymphoma, among whom 16 also received examination by computed tomography (CT) or magnetic resonance imaging (MRI). RESULTS: PET was found to have better diagnostic efficiency than CT or MRI examination and the former provided referential information in all the cases for estimation of the clinical stage, while CT and MRI were helpful in only 66.7% of the cases. FDG uptake as demonstrated by PET helped the assessment of the malignancies and prognosis, according to which treatment adjustment was implemented in 20 cases, resulting in a efficacy rate of 85.7%. CONCLUSION: By integrating morphological and metabolic observations, PET is a superior diagnostic modality in terms of systemic identification, early detection, quantitative analysis, evaluating metabolic function of lesion monitoring therapeutic effect and making prognostic assessment of malignant lymphoma. In spite of the safety of PET examination, false positive results were not eliminated and blood sugar in patients with diabetes might be transiently affected by the tracing agents. PMID- 12527524 TI - [Treatment and etiological analysis of biliary fistula associated with liver hydatidosis: report of 13 cases]. AB - OBJECTIVE: To explore the cause and the treatment of liver hydatidosis complicated by biliary fistula, to reduce or prevent the incidence of the complication. METHODS: From 1982 to 2002, 1 666 patients with liver hydatidosis were treated surgically in our hospital, and a retrospective analysis was conducted in 13 from the cohort, who developed biliary fisltula after the surgeries. RESULTS: The incidence of liver hydatidosis-associated biliary fistula was 0.78% (13/1666), and 8 (61.5%) out of such cases were due to the primary liver hydatidosis, 3 (15.38%) were caused by damage to the small accessory hepatic duct or parahepatic byliary duct, and 2 (7.69%) by the puncture of the needle. CONCLUSION: Biliary fistula often arises from liver hydatidosis itself, and can be partly attributed to erroneous operation on the part of the operators, and prevention of this complication, therefore, is possible through careful operation. PMID- 12527525 TI - [Histological study of the dental implant-bone interface remodeling of submerged and nonsubmerged implants in dogs]. AB - OBJECTIVE: To compare the characteristics of histological reaction at the implant bone interface after submerged and nonsubmerged implant remodeling in dogs. METHODS: Eight adult mongrel dogs were used to establish the animal model for submerged and nonsubmerged implants into the bilateral mandibular sites. At 1, 2, 4, and 12 weeks postoperatively, 2 dogs were sacrificed at each time point for histological observation of the healing and remodeling process at the implant bone interface by HE staining. RESULTS: No distinct difference in the rate of new bone generation and degree of osseointegration was observed between dogs with submerged and nonsubmerged implant at each time point. CONCLUSION: In spite of the bite force imposed on the abutment to some extent during the healing process after nonsubmerged implant plantation, favorable osseointegration similar to that after submerged implants can be achieved. PMID- 12527526 TI - [Construction and application of Access database of colorectal carcinoma cases]. AB - OBJECTIVE: To construct a database using Access software in which clinical information of colorectal cancer cases can be loaded, to facilitate relevant large-sample clinical studies. METHODS: A retrospective study was conducted in 1 374 cases of colorectal carcinoma with surgical treatment between 1975 to 1999 in Nanfang Hospital. According to the National Standards for Pathological Study of Colorectal Carcinoma, an Access2000 database consisting of 1 145 pathologically confirmed colorectal carcinoma cases was established, designated as The Specialized Access Database of Colorectal Carcinoma. RESULTS AND CONCLUSION: The database system has been successfully constructed and operates smoothly, which possesses powerful capacity for information processing of colorectal cancer cases. PMID- 12527527 TI - [Determination of EC50 of ropivacaine and bupivacaine for postoperative epidural analgesia by up-down sequential allocation]. AB - OBJECTIVE: To determine the median effective concentration (EC50) of ropivacaine and bupivacaine for postoperative epidural analgesia, and compare the efficacy of the 2 analgesics. METHODS: Sixty-five patients (ASA I to II) scheduled for elective gynaecological surgeries were randomly assigned into ropivacaine and bupivacaine groups to receive postoperative epidural analgesia with the indicated analgesics (20 ml). The concentration was determined by the response of a previous patient to a higher or lower concentration using double-blinded, up-down sequential allocation. The analgesic efficacy was assessed with 100 mm visual analog pain scores (VAS), and effectiveness was defined as having a VAS 50%) to the antibiotics erythromycin, imipenem, amikacin, SXT, ciprofloxacin, which fell into 4 categories according to their different antibiotic mechanisms. CONCLUSIONS: The prevalence and high MDR rate of MRS isolated form the samples suggest that in cases of infection, identification of the pathogenic bacterium should be routinely performed. Besides controlling MRS infection, importance should also be given to its prevention. PMID- 12527529 TI - [Assessment of variables relative to prethrombotic state after operation in patients with gynecological malignancies]. AB - OBJECTIVE: To study the changes in the variables in patients with gynecological malignancies after operation. METHOD: Platelet alphagranule membrane protein (GMP 140), von Willebrand Factor (vWF), antithrombin III activity (AT-III), protein C dependent partial thromboplastin activation time, plasminogen activity (PLG), the activity of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI), D-Dimer were examined in 20 normal non-pregnant women and 38 patients after operation with gynecological malignancies. RESULTS: vWF, GMP-140, PLG, D-Dimer, and PAI of patients with malignancies before operation were obviously higher than those of the healthy women (P<0.01). After operation the parameters were obviously elevated in the patients (P<0.01). AT-III and partial thromboplastin activation time were significantly reduced in comparison with the healthy subjects (P<0.01). t-PA showed no significant difference between the groups (P>0.05). CONCLUSIONS: Obvious prethrombotic state characterizes the patients with gynecological malignancies after operation. PMID- 12527530 TI - [Treatment of 48 cases of frozen shoulder with manual therapy under brachial plexus anesthesia through a retained tube]. AB - Manual therapy was adopted for treatment of 48 patients with frozen shoulder, under brachial plexus block through a tube that was not withdrawn until 1 to 2 weeks after the initiation of the treatment course. Satisfactory results were achieved in most of the patients after the treatment, indicating the safety and feasibility of brachial plexus block for pain relief through long-term retention of the tube. PMID- 12527531 TI - [Application of nitrocellulose membrane in cell culture and pathological techniques]. AB - OBJECTIVE: To provide a new supporting medium for carrying out cell culture and histological staining. METHOD: LoVo cell line was cultured using nitrocellulose membrane as the culture medium, in parallel with the same cell culture on the coverglass which served as control. RESULTS: The cells cultured on nitrocellulose membrane showed no visible difference from those cultured on coverglasses in terms of cell growth, morphology, and the structure displayed after staining. No toxic effect was observed due to the application of nitrocellulose membrane. CONCLUSIONS: Nitrocellulose membrane after treatment has such merit as good lucidity and stability without toxicity on the cells. Having also good affinity with the cells, nitrocellulose membrane can offer an promising alternative for glass as cell culture medium. PMID- 12527532 TI - [Usage of participles, compound words and nouns as attributives in English meclical papers]. AB - In this paper, the usage of participles and compound words as attributives in English medical papers is discussed with also regard to the application of nouns in groups, specifically for the benefit of Chinese authors. PMID- 12527533 TI - Advances in psychotherapy for mental illness in late life. PMID- 12527534 TI - Psychotherapy research with older populations. PMID- 12527535 TI - Guidelines for conducting geropsychotherapy research. AB - Geropsychotherapy researchers have established specific methods that improve the reliability and generalizability of the data from this research. To date, there has been little formal dissemination of these methods. The authors present guidelines for the optimal conduct of psychotherapy research in older adults, which include selection of age-appropriate psychotherapies and control conditions, use of consumer-based methods for recruitment, evaluation of age related treatment processes and outcomes, and adjusting the research design to accommodate age-specific life events and provide examples of how each guideline was used in their psychotherapy studies. Psychotherapy research with older adults has benefited from methodological advances that improve our ability to ascertain the impact of psychotherapy on late-life disorders. However, the field is still in need of better outcome and process measures, methods for measuring the therapeutic content of non-psychotherapy encounters, and methods for determining the impact of choice of treatment on outcome. PMID- 12527536 TI - Functional adaptation skills training (FAST): a pilot psychosocial intervention study in middle-aged and older patients with chronic psychotic disorders. AB - OBJECTIVE: Developing behavioral interventions to improve functioning of older patients with schizophrenia and other chronic psychoses has the potential to significantly increase the patients' independence and quality of life. METHODS: The authors evaluated a psychosocial intervention designed to improve everyday living skills of middle-aged and older outpatients with very chronic psychotic disorders (mean duration of illness: 21 years). Forty patients who resided in board-and-care facilities were randomly assigned to either a 24-session functional adaptation skills training (FAST) group therapy program targeting problem areas identified in previous work as being problematic for this population (e.g., using public transportation) or treatment-as-usual. Almost all the participants also received antipsychotics. RESULTS: Compared with the patients randomized to the treatment-as-usual condition, FAST-treated patients' performance on everyday living skills improved significantly immediately post intervention and was still significantly better at a 3-month maintenance follow up period. There was no significant change in psychopathology. CONCLUSION: Results suggest that older patients with longstanding psychotic disorders may benefit from participation in this skills-training program. PMID- 12527537 TI - Standard and enhanced cognitive-behavior therapy for late-life generalized anxiety disorder: two pilot investigations. AB - OBJECTIVE: Although cognitive-behavior therapy (CBT) is an efficacious treatment for generalized anxiety disorder (GAD) in younger adults, little is known about its efficacy in older patients. Investigations to date have tested group-format or otherwise nonstandard versions of CBT. The studies described here are, to our knowledge, the first to test the efficacy of individual-format CBT administered in a mental health clinic for treatment of late-life GAD. METHODS: Study 1 tested a standard version, and Study 2 tested an enhanced version (ECBT) that included learning and memory aids designed to make the therapy more effective with elderly patients (e.g., homework reminder and troubleshooting calls, weekly review of all concepts and techniques). RESULTS: Study 1 CBT participants showed significant improvement on GAD severity ratings. Study 2 ECBT participants showed significant improvement on two self-report measures, rates of posttreatment GAD, and GAD severity ratings. ECBT resulted in improvement on more measures and yielded larger effect sizes than standard CBT, when each was compared against a wait-list control group. CONCLUSION: Results of these pilot studies suggest that standard individual-format CBT may not be optimally effective for treating late-life GAD; thus, ECBT should be tested further in randomized trials. PMID- 12527538 TI - Dialectical behavior therapy for depressed older adults: a randomized pilot study. AB - OBJECTIVE: Although there is evidence for the efficacy of antidepressants and for some individual and group psychotherapy interventions for depressed older adults, a significant number of these do not respond to treatment. Authors assessed the benefits of augmenting medication with group psychotherapy. METHODS: They randomly assigned 34 (largely chronically) depressed individuals age 60 and older to receive 28 weeks of antidepressant medication plus clinical management, either alone (MED) or with the addition of dialectical behavior therapy skills-training and scheduled telephone coaching sessions (MED+DBT). RESULTS: Only MED+DBT showed significant decreases on mean self-rated depression scores, and both treatment groups demonstrated significant and roughly equivalent decreases on interviewer rated depression scores. However, on interviewer-rated depression, 71% of MED+DBT patients were in remission at post-treatment, in contrast to 47% of MED patients. At a 6-month follow-up, 75% of MED+DBT patients were in remission, compared with only 31% of MED patients, a significant difference. Only patients receiving MED+DBT showed significant improvements from pre- to post-treatment on dependency and adaptive coping that are proposed to create vulnerability to depression. CONCLUSION: Results from this pilot study suggest that DBT skills training and telephone coaching may offer promise to effectively augment the effects of antidepressant medication in depressed older adults. PMID- 12527539 TI - Problem-solving therapy versus supportive therapy in geriatric major depression with executive dysfunction. AB - OBJECTIVE: The authors compared the efficacy of problem-solving therapy (PST) and supportive therapy (ST) in a group of elderly subjects with impairment in executive functions. This group was targeted because it has been shown to be at the risk for poor response to pharmacotherapy. METHODS: A total of 25 elderly subjects with major depression and abnormal scores in initiation/perseveration and response inhibition tasks were randomly assigned to receive weekly sessions of PST or ST for 12 weeks. The subjects were systematically evaluated by raters blind to the study hypotheses. RESULTS: PST was more effective than ST in leading to remission of depression, fewer post-treatment depressive symptoms, and less disability. A substantial part of the change in depression and disability was explained by the subjects' improvement of skills in generating alternatives and in decision-making. CONCLUSION: This preliminary study suggests that PST is effective in reducing depressive symptoms and disability in elderly patients with major depression and executive dysfunction. If these findings are confirmed, PST may become an important therapeutic alternative for a patient population who may otherwise remain symptomatic and disabled. PMID- 12527540 TI - The relationship between homework compliance and treatment outcomes among older adult outpatients with mild-to-moderate depression. AB - OBJECTIVE: The authors extend previous research on homework in psychotherapy by examining the relationship between homework compliance and therapeutic outcome among depressed older adult outpatients (N = 63), addressing previous limitations by using session-by-session therapist ratings of homework compliance and including both interviewer ratings and patient self-reports of outcomes. METHODS: Patients were participants in a randomized clinical trial evaluating the efficacy of desipramine versus cognitive/behavioral therapy-alone (C/B-Alone) versus a combination of the two (Combined). Given the current study's focus on homework compliance, only patients assigned to conditions with assigned homework in the clinical trial (i.e., C/B-Alone and Combined conditions) were included. RESULTS: Results of hierarchical regression analyses indicated that homework compliance contributed significantly to posttreatment outcome as measured by both interviewer-administered and patient self-report measures of depression. A separate series of ANOVAs also found significant differences in pre-posttreatment change between patients scoring above and below the median of reported homework compliance. Findings were similar for patients in the C/B-Alone and Combined conditions. CONCLUSION: The study's results call for additional research on issues related to homework compliance with older adult patients. PMID- 12527541 TI - In-home respite intervention reduces plasma epinephrine in stressed Alzheimer caregivers. AB - OBJECTIVE: Some excess morbidity and mortality in Alzheimer caregivers (CGs) may be related to chronic activation of the sympatho-adrenal-medullary (SAM) system. Authors tested the efficacy of an in-home respite intervention to reduce peripheral markers of SAM activation and psychological distress in spousal caregivers of patients with Alzheimer disease. METHODS: Caregivers were classified as Vulnerable (n = 27) or Non-Vulnerable (n = 28). Vulnerable CGs were those with a severe mismatch between caregiving demand and help received in the preceding 6 months. CGs had plasma catecholamine levels sampled at rest and in response to a stressor (speech task) before and 1 month after a 2-week in-home respite intervention. Self-reported symptoms of anxiety and depression were also obtained. RESULTS: ANOVA revealed a group x treatment interaction: At the 1-month follow-up, plasma epinephrine declined significantly in the vulnerable caregivers who received respite, but rose in those who were wait-listed. No effect was found for norepinephrine, heart rate, blood pressure, or psychological symptoms. CONCLUSION: Findings suggest that an in-home respite program may reduce SAM activation independent of psychological symptoms. To the extent that sympathetic activation mediates pathophysiological events, these results suggest an approach that may reduce morbidity and mortality in certain caregivers. PMID- 12527542 TI - Is the APOE epsilon4 genotype associated with higher hospital costs among elderly patients? AB - OBJECTIVE: The apolipoprotein epsilon4 (APOE epsilon4) genotype is associated with a number of adverse health outcomes. The authors assessed whether the epsilon4 genotype was associated with higher hospital costs on the basis of data from 1,999 white or black respondents to the Duke Established Population for Epidemiological Studies of the Elderly who consented to be genotyped in 1992 1993. METHODS: They measured hospital costs, using the amount paid by Medicare for hospitalizations from 1992 to 1997. RESULTS: Persons with the epsilon4 genotype did not have higher costs than those who were epsilon4-negative. The highest costs were observed for those who had missing epsilon4 genotype. CONCLUSION: The epsilon4 genotype is not a significant predictor of hospital costs, and so would not be a good risk adjustor for purposes such as setting reimbursement rates for Medicare risk plans. PMID- 12527543 TI - The epidemiology of psychosis in dementia. AB - OBJECTIVE: Authors compared delusions, hallucinations, and misidentification delusions in Alzheimer disease (AD) and vascular dementia (VaD) patients. METHODS: The authors report data on the prevalence, severity, clinical, and demographic associations of these symptoms in a population sample of 260 persons with dementia, examined with the Neuropsychiatric Inventory. RESULTS: The primary finding was that there was no difference in psychosis as a whole, or in delusions and hallucinations, between AD and VaD. Also, in AD, female gender appeared to be a risk factor for delusions; subjects in an earlier stage of dementia showed fewer delusions. CONCLUSION: The profile of delusions and hallucinations seen is different from that seen in schizophrenia, further supporting the hypothesis that AD-associated psychosis is a distinct phenomenological syndrome. PMID- 12527544 TI - Cognitive-behavior therapy for late-life generalized anxiety disorder in primary care: preliminary findings. AB - OBJECTIVE: The authors present data from a pilot research program initiated to develop, refine, and test the outcomes of CBT-GAD/PC, a version of cognitive behavior therapy (CBT) that targets the needs of older adults with generalized anxiety disorder (GAD) in primary care (PC). METHODS: The study involved a small, randomized clinical trial comparing the impact of CBT-GAD/PC to usual care (UC) in a sample of 12 older medical patients with GAD. RESULTS: Outcome data suggested significant improvements in worry and depression after CBT-GAD/PC, relative to UC. CONCLUSION: Authors discuss results in terms of the "real-world" applicability of this treatment for late-life GAD, and present implications for future research. PMID- 12527545 TI - The value of maintenance interpersonal psychotherapy (IPT) in older adults with different IPT foci. AB - OBJECTIVE/METHODS: The authors examined recurrence rates of major depression in elderly subjects with different foci of interpersonal psychotherapy (IPT), who were treated for up to 3 years with either monthly maintenance IPT and pill placebo or with monthly clinical management and pill placebo. RESULTS: Among subjects with an IPT focus on role conflict, a greater proportion of those treated with maintenance-IPT survived for 3 years without recurrence than those treated with placebo/clinical management. Median time to recurrence was 68.9 weeks in IPT-treated patients versus 16.3 weeks for patients in clinical management. Subjects with an IPT focus on abnormal grief or role transition demonstrated no effect differential for maintenance IPT and pill-placebo on recurrence prevention over supportive clinical management and pill-placebo. CONCLUSION: If replicated in a larger sample, these findings have important implications for ongoing case-management decisions. PMID- 12527546 TI - Cognitive and neurologic predictors of functional impairment in vascular dementia. AB - OBJECTIVE: The purpose of this study is to investigate associations between executive dysfunction, neuroimaging findings, and functional impairment in patients with vascular dementia (VaD). METHODS: Twenty-nine VaD patients completed the Dementia Rating Scale and underwent MRI scanning to generate quantitative ratings of subcortical hyperintensities (SH) and cortical volume. Patients' caregivers completed items from the Lawton and Brody Activities of Daily Living Questionnaire, designed to measure instrumental activities of daily living (IADLs). The authors hypothesized that performance on the Initiation/Perseveration (IP) subscale, a measure of executive abilities, and SH would significantly predict levels of IADLs. RESULTS: A hierarchical multiple regression analysis revealed that IP and SH accounted for 42% of the variance in IADLs; IP alone accounted for 28%, and SH accounted for 14% beyond the contribution made by IP. CONCLUSION: Findings indicate that specific cognitive and neuropathological factors are associated with functional impairment in VaD. PMID- 12527547 TI - The pharmaceutical industry in the nursing home: no such thing as a free lunch. PMID- 12527549 TI - Coma with accidental single dose of an atypical neuroleptic in a patient with Lewy Body dementia. PMID- 12527550 TI - Patient choice and end-of-life decisions. PMID- 12527551 TI - Impaired ability of the Na+/Ca2+ exchanger from the Dahl/Rapp salt-sensitive rat to regulate cytosolic calcium. AB - We previously cloned Na(+)/Ca(2+) exchanger (NCX1) from mesangial cells of salt sensitive (SNCX = NCX1.7) and salt-resistant (RNCX = NCX1.3) Dahl/Rapp rats. The abilities of these isoforms to regulate cytosolic Ca(2+) concentration ([Ca(2+)](i)) were assessed in fura 2-loaded OK cells expressing the vector (VOK), RNCX (ROK), and SNCX (SOK). Baseline [Ca(2+)](i) was 98 +/- 20 nM (n = 12) in VOK and was significantly lower in ROK (44 +/- 5 nM; n = 12) and SOK (47 +/- 13 nM; n = 12) cells. ATP at 100 microM increased [Ca(2+)](i) by 189 +/- 55 nM (n = 12), 21 +/- 9 nM (n = 12), and 69 +/- 18 nM (n = 12) in VOK, ROK, and SOK cells, respectively. ATP (1 mM) or bradykinin (0.1 mM) caused large increases in [Ca(2+)](i) and ROK but not SOK cells were much more efficient in reducing [Ca(2+)](i) back to baseline levels. Parental Sprague-Dawley rat mesangial cells express both RNCX (SDRNCX) and SNCX (SDSNCX). SDRNCX and RNCX are identical at every amino acid residue, but SDSNCX and SNCX differ at amino acid 218 where it is isoleucine in SDSNCX and not phenylalanine. OK cells expressing SDSNCX (SDSOK) reduced ATP (1 mM)-induced [Ca(2+)](i) increase back to baseline at a rate equivalent to that for ROK cells. PKC downregulation significantly attenuated the rate at which ROK and SDSOK cells reduced ATP-induced [Ca(2+)](i) increase but had no effect in SOK cells. The reduced efficiency of SNCX to regulate [Ca(2+)](i) is attributed, in part, to the isoleucine-to-phenylalanine mutation at amino acid 218. PMID- 12527552 TI - Localization of human equilibrative nucleoside transporters, hENT1 and hENT2, in renal epithelial cells. AB - Nucleoside transporters are important in the disposition of nucleosides and nucleoside analogs in the kidney. Two human equilibrative nucleoside transporters have been cloned and characterized, hENT1 and hENT2. The primary goal of this study was to localize these transporters in polarized renal epithelia. hENT1 and hENT2 were tagged with green fluorescence protein, stably expressed in renal epithelial cells, and localized by immunofluorescence and functional analysis. Our data demonstrated that both transporters are expressed on the basolateral membrane. hENT1 is also present on the apical membrane. Additionally, we examined the importance to basolateral targeting of two COOH-terminal targeting motifs: a RXXV motif for hENT1 and a dileucine repeat for hENT2. Neither motif appeared to affect targeting, but the dileucine repeat was implicated in surface expression of hENT2. In addition, a splice variant of hENT2 was identified that is predicted to result in a 156-residue COOH-terminal truncation. This variant had a tissue distribution similar to wild-type hENT2 but was retained intracellularly. These data suggest that hENT1 and hENT2 on the basolateral membrane function with concentrative nucleoside transporters on the apical membrane to mediate active reabsorption of nucleosides within the kidney. PMID- 12527553 TI - Angiotensin II induces apoptosis in renal proximal tubular cells. AB - ANG II has been demonstrated to play a role in the progression of tubulointerstial injury. We studied the direct effect of ANG II on apoptosis of cultured rat renal proximal tubular epithelial cells (RPTECs). ANG II promoted RPTEC apoptosis in a dose- and time-dependent manner. This effect of ANG II was attenuated by anti-transforming growth factor (TGF)-beta antibody. Moreover, TGF beta triggered RPTEC apoptosis in a dose-dependent manner. ANG II also enhanced RPTEC expression of Fas and Fas ligand (FasL); furthermore, anti-FasL antibody attenuated ANG II-induced RPTEC apoptosis. In addition, ANG II increased RPTEC expression of Bax, a cell death protein. Both ANG II type 1 (AT(1)) and type 2 (AT(2)) receptor blockers inhibited ANG II-induced RPTEC apoptosis. SB-202190, an inhibitor of p38 MAPK phosphorylation, and caspase-3 inhibitor also attenuated ANG II-induced RPTEC apoptosis. ANG II enhanced RPTEC heme oxygenase (HO)-1 expression. Interestingly, pretreatment with hemin as well as curcumin (inducers of HO-1) inhibited the ANG II-induced tubular cell apoptosis; conversely, pretreatment with zinc protoporphyrin, an inhibitor of HO-1 expression, promoted the effect of ANG II. These results suggest that ANG II-induced apoptosis is mediated via both AT(1) and AT(2) receptors through the generation of TGF-beta, followed by the transcription of cell death genes such as Fas, FasL, and Bax. Modulation of tubular cell expression of HO-1 has an inverse relationship with the ANG II-induced tubular cell apoptosis. PMID- 12527554 TI - Role of PKC and calcium in modulation of effects of angiotensin II on sodium transport in proximal tubule. AB - It has been well documented that low concentrations of ANG II (10(-11) to 10(-10) M) stimulate, whereas high concentrations of ANG II (10(-8) to 10(-5) M) inhibit Na(+) transport in proximal tubules of rat and rabbit kidneys. Measured ANG II concentration in proximal tubular fluid is in the nanomolar range. In the present study, we investigated the role of PKC, intracellular Ca(2+), and cAMP in modulating the effects of luminal ANG II on Na(+) absorption by microperfusion techniques in rabbit superficial segment of proximal tubules in vitro. We confirmed that ANG II (10(-9) M) had no change on fluid absorption (J(v)); however, fluid absorption increased significantly when 10(-9) M ANG II and 3,4,5 trimethoxybenzoic acid-8-(diethylamino)octyl ester (TMB-8), a blocker of intracellular calcium mobilization, were added together. In contrast, ANG II significantly decreased J(v) when PKC was inhibited. When 10(-9) M ANG II was present together with 1-(5-isoquinolinesulfonyl)-2-mehtylpiperazine and TMB-8, no significant change of J(v) occurred. Inhibition of endogenous cAMP activity by a PKA inhibitor did not change either basal or ANG II-stimulated fluid absorption. Our results indicate that ANG II regulates Na(+) absorption by a cAMP-independent mechanism and that PKC and intracellular calcium both play a critical role in modulating the effects of physiological concentration of ANG II on proximal tubule transport. Balance between these two cytosolic messengers modulates the effects of ANG II on fluid absorption in the proximal tubule. PMID- 12527555 TI - Determinants of GFR depression in early membranous nephropathy. AB - We evaluated the glomerular filtration rate (GFR) in 34 subjects with membranous nephropathy (MN) of new onset. We used physiological techniques to measure GFR, renal plasma flow, and oncotic pressure and computed a value for the two-kidney ultrafiltration coefficient (K(f)). A morphometric analysis of glomeruli in the diagnostic biopsy permitted computation of the single-nephron ultrafiltration coefficient (SNK(f)). MN subjects were divided into two groups: moderate or severe, according to whether GFR was depressed by less or more than 50%. SNK(f) was subnormal but similar in moderate and severe MN. In contrast, two-kidney K(f) was significantly more depressed in severe than in moderate MN. We estimated the total number of functioning glomeruli (N(g)) by dividing two-kidney K(f) by SNK(f). Whereas mean N(g) was similar in controls and moderate MN (1.5 and 1.4 1.7 x 10(6), respectively), it was significantly lower in severe MN (0.5 x 10(6)). This degree of glomerulopenia was not reflected in the rate of global sclerosis. We conclude that a combination of depressed SNK(f) (due to foot process broadening) and profound glomerulopenia accounts for GFR depression of >50% early in the course of MN. The cause of the glomerulopenia remains to be elucidated. PMID- 12527556 TI - Disruption of F-actin stimulates hypertonic activation of the BGT1 transporter in MDCK cells. AB - Many membrane transport systems are altered by changes in the state of the actin cytoskeleton. Although an intact microtubule network is required for hypertonic activation of the betaine transporter (BGT1), the possible role of the actin cytoskeleton is unknown. BGT1 function in Madin-Darby canine kidney cell monolayers was assessed as Na(+)-dependent uptake of GABA, following disassembly of F-actin by cytochalasin D (1.0 microM) or latrunculin A (0.6 microM). Both drugs significantly increased (P < 0.001) the activation of BGT1 transport by 24 h hypertonicity (500 mosmol/kgH(2)O). In contrast, the hypertonic upregulation of Na(+)-dependent alanine uptake remained unaltered by cytochalasin D. Disruption of F-actin did not interfere with downregulation of BGT1 transport when cells were transferred from hypertonic to isotonic medium. Immunofluorescence staining revealed colocalization of BGT1 and F-actin at the plasma membrane of hypertonic cells. Surface biotinylation revealed no major change in BGT1 protein abundance after cytochalasin D action, suggesting that stimulation of hypertonic activation of BGT1 transport is due to increased activity of existing BGT1 transporters. PMID- 12527557 TI - Disruption of bladder epithelium barrier function after spinal cord injury. AB - Neural-epithelial interactions are hypothesized to play an important role in bladder function. We determined whether spinal cord injury (SCI) altered several indicators of urinary bladder epithelium barrier function, including continuity of the surface umbrella cell layer, transepithelial resistance (TER), and urea and water permeability. Within 2 h of SCI, significant changes in uroepithelium were noted, including disruption of the surface umbrella cells and an approximately 50% decrease in TER. By 24 h, TER reached a minimum and was accompanied by significant increases in water and urea permeability. Regeneration of the surface uroepithelium was accomplished by 14 days after SCI and was accompanied by a return to normal TER and urea and water permeabilities. This early disruption of the uroepithelial permeability and accompanying changes in uroepithelial morphology were prevented by pretreatment with hexamethonium (a blocker of ganglion transmission), indicating involvement of sympathetic or parasympathetic input to the urinary bladder. In addition, prior treatment with capsaicin worsened the effect of SCI on uroepithelial permeability, suggesting that capsaicin-sensitive afferents may play a protective role in the process. These results demonstrate that SCI results in a significant disruption of the urinary bladder uroepithelium and that these changes may be mediated in part by an interaction with bladder nerves. PMID- 12527558 TI - Arginine, ornithine, and proline interconversion is dependent on small intestinal metabolism in neonatal pigs. AB - We have previously shown that arginine deficiency is exacerbated by the removal of dietary proline in orally, but not parenterally, fed piglets. Therefore, we hypothesized that the net interconversions of proline, ornithine, and arginine primarily occur in the small intestine of neonatal piglets. Ten intragastrically fed piglets received either intraportal (IP) or intragastric (IG) primed, constant infusions of [guanido-(14)C]arginine and [U-(14)C]ornithine + [2,3 (3)H]proline. By infusing amino acid isotopes via the stomach compared with the portal vein, we isolated small intestinal first-pass metabolism in vivo. During IP infusion, fractional net conversions (%) from proline to ornithine (0), ornithine to arginine (11 +/- 6), and ornithine to proline (5 +/- 1) were lower (P < 0.05) than during IG infusion (39 +/- 8, 18 +/- 6, and 42 +/- 12, respectively); we speculate that these data are due to the localization of ornithine aminotransferase to the gut. The balance of these conversions indicated a large synthesis of arginine (70.0 micromol. kg(-1). h(-1)) by the gut, with a corresponding degradation of ornithine (70.8 micromol. kg(-1). h(-1)) and no change in proline balance. Gut synthesis of arginine from proline (48.1 micromol. kg(-1). h(-1)) was 50% of its requirement, whereas proline synthesis from arginine (33.0 micromol. kg(-1). h(-1)) amounted to 10% of its requirement. Overall, arginine synthesis is more dependent on the gut than proline synthesis. In situations in which gut metabolism is compromised, such as during parenteral nutrition or gastrointestinal disease, arginine and proline are individually indispensable because their biosyntheses are negligible. PMID- 12527559 TI - BMP-2 inhibits proliferation of human aortic smooth muscle cells via p21Cip1/Waf1. AB - Bone-morphogenetic proteins (BMP)-2 and -7, multifunctional members of the transforming growth factor (TGF)-beta superfamily with powerful osteoinductive effects, cause cell cycle arrest in a variety of transformed cell lines by activating signaling cascades that involve several cyclin-dependent kinase inhibitors (CDKIs). CDKIs in the cip/kip family, p21(Cip1/Waf1) and p27(Kip1), have been shown to negatively regulate the G1 cyclins and their partner cyclin dependent kinase proteins, resulting in BMP-mediated growth arrest. Bone morphogens have also been associated with antiproliferative effects in vascular tissue by unknown mechanisms. We now show that BMP-2-mediated inhibition of platelet-derived growth factor (PDGF)-stimulated human aortic smooth muscle cell (HASMC) proliferation is accompanied by increased levels of p21 protein. Antisense oligodeoxynucleotides specific for p21 attenuate BMP-2-induced inhibition of proliferation when transfected into HASMCs, demonstrating that BMP 2 inhibits PDGF-stimulated proliferation of HASMCs through induction of p21. Whether p21-mediated induction of cell cycle arrest by BMP-2 sets the stage for osteogenic differentiation of vascular smooth muscle cells, ultimately leading to vascular mineralization, remains to be investigated. PMID- 12527560 TI - Diagnosis and management of lung cancer: ACCP evidence-based guidelines. American College of Chest Physicians. PMID- 12527561 TI - Overview of methodology for lung cancer evidence review and guideline development. AB - A multidisciplinary panel was convened by the American College of Chest Physicians (ACCP) to develop clinical practice guidelines for lung cancer prevention, diagnosis, and treatment. The ACCP Expert Panel on Lung Cancer Guidelines produced 20 guidelines, each related to a distinct set of management decisions. This article describes the approach used to develop the guidelines, including identifying, evaluating, and synthesizing evidence, assessing the strength of evidence pertinent to individual guidelines, and grading guideline recommendations. PMID- 12527562 TI - Assessment of the scope and quality of clinical practice guidelines in lung cancer. AB - STUDY OBJECTIVES: To provide an evidence-based background for developing the American College of Chest Physicians (ACCP) lung cancer guidelines, a systematic review of the literature was performed to identify published lung cancer guidelines and evaluate their quality. DESIGN, SETTING, AND PARTICIPANTS: A systematic search was performed for relevant literature from MEDLINE, Cancerlit, CINAHL, HealthStar, the Cochrane Library, and the National Guidelines Clearinghouse published from January 1989 to July 2001. MEASUREMENT AND RESULTS: From 369 citations, 51 relevant guidelines were identified. Each guideline was evaluated by at least four reviewers using the Appraisal of Guidelines for Research and Evaluation (AGREE) instrument and was coded for clinical topics covered. The recommendations included in each guideline also were abstracted. Of the 51 guidelines evaluated, 27 (53%) were evidence-based. Clinical topics identified by the ACCP for their guideline effort each were represented by at least one existing guideline. Of the 880 clinical recommendations abstracted from the guidelines, only 253 (29%) were evidence-based. The AGREE instrument rates guidelines along six domains. As a group, the guidelines performed well in the scope and purpose domain, with only six guidelines (12%) scoring < 50%. For the remaining domains, however, the guidelines did not perform as well, as follows: for stakeholder involvement, 41 guidelines (80%) scored < 50%; for rigor of development, 29 guidelines (57%) scored < 50%; for clarity and presentation, 17 guidelines (33%) scored < 50%; for applicability, 46 guidelines (90%) scored < 50%; and for editorial independence, 47 guidelines (92%) scored < 50%. After considering the domain scores, the reviewers recommended only 19 of the guidelines (37%). CONCLUSIONS: All major clinical lung cancer topics are covered by at least one guideline, but no single guideline addresses all areas. Furthermore, although existing guidelines may accurately reflect clinical practice, most performed poorly when evaluated for quality. Future guideline efforts that address each item of the AGREE instrument would add substantially to the literature. PMID- 12527563 TI - Epidemiology of lung cancer. AB - In the United States, lung cancer remains the leading cause of cancer death in both men and women even though an extensive list of risk factors has been well characterized. Far and away the most important cause of lung cancer is exposure to tobacco smoke through active or passive smoking. The reductions in smoking prevalence in men that occurred in the late 1960s through the 1980s will continue to drive the lung cancer mortality rates downward in men during the first portion of this century. This favorable trend will not persist unless further reductions in smoking prevalence are achieved. PMID- 12527564 TI - Prevention of lung cancer: summary of published evidence. AB - STUDY OBJECTIVES: To describe empiric research related to lung cancer prevention strategies, including chemoprevention aimed at reducing lung cancer incidence and various smoking avoidance and cessation interventions aimed at reducing smoking rates. DESIGN, SETTING, AND PARTICIPANTS: Systematic searches of MEDLINE, HealthStar, and Cochrane Library databases to July 2001 and print bibliographies. For chemoprevention studies, we considered only randomized controlled trials (RCTs) with lung cancer incidence as an end point. For studies of smoking avoidance or cessation, we selected systematic reviews and meta-analyses, and searched for individual RCTs only where high-quality and current reviews and meta analyses were not available. MEASUREMENT AND RESULTS: Chemoprevention of lung cancer has been studied in five RCTs of primary prevention, no RCTs of secondary prevention, and five RCTs of tertiary prevention. None of these trials has shown evidence for efficacy of any agents tested, including retinol (vitamin A), beta carotene, N-acetylcysteine, and selenium. There is a great deal of evidence about a wide variety of clinician-based and community-based efforts at smoking avoidance or cessation. Certain approaches have been shown to be effective (eg, mass media public education campaigns, direct restrictions on smoking, clinician based approaches ranging from brief clinician advice to more in-depth sessions, and "life-skills training" in schools). Some approaches have intermediate or short-term effectiveness (ie, youth access restrictions and school-based interventions), and others have been shown to be ineffective (ie, acupuncture and provider education) or have been insufficiently studied (ie, provider feedback). CONCLUSIONS: There are no agents that have been proven to be effective for preventing lung cancer. Several clinician-based and community-based interventions show promise for reducing lung cancer incidence through smoking avoidance and prevention. PMID- 12527565 TI - Lung cancer prevention: the guidelines. AB - Lung carcinogenesis is a chronic and multi-step process resulting in malignant lung tumors. This progression from normal to neoplastic pulmonary cells or tissues could be arrested or reversed through pharmacologic treatments, which are known as cancer chemoprevention. These therapeutic interventions should reduce or avoid the clinical consequences of lung cancer by treating early neoplastic lesions before the development of clinically evident signs or symptoms of malignancy. Preclinical, clinical, and epidemiologic findings relating to different classes of candidate chemopreventive agents provide strong support for lung cancer prevention as an attractive therapeutic strategy. Smoking prevention and smoking cessation represent an essential approach to reduce the societal impact of tobacco carcinogenesis. However, even if all the goals of the national antismoking efforts were met, there still would be a large population of former smokers who would be at increased risk for lung cancers. Lung cancer also can occur in those persons who never have smoked. This article focuses on what is now known about pharmacologic strategies for lung cancer prevention. Randomized clinical trials using beta-carotene, retinol, isotretinoin or N-acetyl-cysteine did not show benefit for primary and tertiary lung cancer prevention. There is also evidence that the use of beta-carotene and isotretinoin for lung cancer chemoprevention in high-risk individuals may increase the risk for lung cancer, especially in individuals who continue to smoke. There is a need for relevant in vitro models to identify pathways that activate chemopreventive effects in the lung. An improved understanding of cancer prevention mechanisms should aid in the design of clinical trials and in the validation of candidate chemopreventive targets as well as the discovery of new targets. Until such studies are completed, no agent or combination of agents should be used for lung cancer prevention outside of a clinical trial. PMID- 12527566 TI - Screening for lung cancer: a review of the current literature. AB - STUDY OBJECTIVES: To review the available data on the early detection of lung cancer, with a focus on three technologies: chest x-ray (CXR), sputum cytology, and low-dose CT (LDCT) scanning. DESIGN, SETTING, PARTICIPANTS: Review of published clinical studies of early detection technologies. The best available evidence on each topic was selected for analysis. Randomized trials were used to evaluate CXR and sputum cytology. Cohort studies, as well as studies providing evidence regarding rates of overdiagnosis and efficacy of initial treatment, were considered in evaluation of LDCT. Study design and results were summarized in evidence tables. Statistical analyses of combined data were not performed. MEASUREMENT AND RESULTS: Five randomized trials of CXR with or without sputum cytology have been conducted, each which reports disease-specific mortality as well as other end points. None of these studies provide support for the use of either CXR or sputum cytology for the early detection of lung cancer in asymptomatic individuals. Eight completed and ongoing trials of LDCT were identified. All of these studies report the frequency and stage distribution of lung cancers found during initial ("prevalence") screening, and several studies also report rates of detection at the time of annual follow-up. No outcome data on survival or treatment are available. A number of studies support the hypothesis of "overdiagnosis"--that some lung cancers detected by LDCT may behave in an indolent manner. CONCLUSIONS: The use of either CXR or sputum cytology for the early detection of lung cancer is not supported by the published evidence. The evidence for LDCT appears promising, in that the technology typically identifies lung cancer at an early stage, although corollary studies suggest that these findings in isolation may be misleading. Further high-quality research is needed to better define the role of LDCT in the evaluation of asymptomatic high risk individuals. PMID- 12527567 TI - Screening for lung cancer: the guidelines. AB - Although virtually all individuals with advanced lung cancer succumb to the disease, a substantial portion of individuals diagnosed at an earlier stage can be cured. This dichotomy has provoked interest in lung cancer screening. To date, randomized controlled trials of chest x-ray and sputum cytology have failed to demonstrate that screening with either modality decreases lung cancer mortality; neither of these technologies can be recommended. Early studies of lung cancer screening with low-dose CT (LDCT) appear promising; however, only data from observational studies are available. We recommend that individuals should only be screened with LDCT in the context of well-designed clinical trials. PMID- 12527568 TI - The solitary pulmonary nodule. AB - More than 150,00 patients a year present to their physicians with the diagnostic dilemma of a solitary pulmonary nodule (SPN) found either on chest radiography or chest CT. A thoughtful and timely workup of this finding is essential if lung cancer is to be recognized early and the chance for cure optimized. Based on the literature to date, recommendations are made for appropriate imaging modalities and diagnostic testing, as well as indications for obtaining preoperative tissue diagnosis for the patient with an SPN. PMID- 12527569 TI - Initial evaluation of the patient with lung cancer: symptoms, signs, laboratory tests, and paraneoplastic syndromes. AB - This chapter describes the components of the initial evaluation for a patient either suspected or known to have lung cancer. The components of the initial evaluation are based on the recognized manifestations of localized lung cancer, ie, symptoms referable to the primary tumor, intrathoracic spread of lung cancer, and patterns of metastatic dissemination. Features of the history and physical signs may be useful indicators of the extent of disease. A standardized evaluation, relying on symptoms, signs, and routinely available laboratory tests, can serve as a useful screen for metastatic disease. Also described are the common features of the various paraneoplastic syndromes associated with lung cancer. PMID- 12527570 TI - The physiologic evaluation of patients with lung cancer being considered for resectional surgery. AB - The preoperative physiologic assessment of a patient being considered for surgical resection of lung cancer must consider the immediate perioperative risks from comorbid cardiopulmonary disease, the long-term risks of pulmonary disability, and the threat to survival due to inadequately treated lung cancer. As with any planned major operation, especially in a population predisposed to atherosclerotic cardiovascular disease by cigarette smoking, a cardiovascular evaluation is an important component in assessing perioperative risks. Measuring the FEV(1) and the diffusing capacity of the lung for carbon monoxide (DLCO) measurements should be viewed as complementary physiologic tests for assessing risk related to pulmonary function. If there is evidence of interstitial lung disease on radiographic studies or undue dyspnea on exertion, even though the FEV(1) may be adequate, a DLCO should be obtained. In patients with abnormalities in FEV(1) or DLCO identified preoperatively, it is essential to estimate the likely postresection pulmonary reserve. The amount of lung function lost in lung cancer resection can be estimated by using either a perfusion scan or the number of segments removed. A predicted postoperative FEV(1) or DLCO < 40% indicates an increased risk for perioperative complications, including death, from lung cancer resection. Exercise testing should be performed in these patients to further define the perioperative risks prior to surgery. Formal cardiopulmonary exercise testing is a sophisticated physiologic testing technique that includes recording the exercise ECG, heart rate response to exercise, minute ventilation, and oxygen uptake per minute, and allows calculation of maximal oxygen consumption (.VO(2)max). Risk for perioperative complications can generally be stratified by .VO(2)max. Patients with preoperative .VO(2)max > 20 mL/kg/min are not at increased risk of complications or death; .VO(2)max< 15 mL/kg/min indicates an increased risk of perioperative complications; and patients with .VO(2)max < 10 mL/kg/min have a very high risk for postoperative complications. Alternative types of exercise testing include stair climbing, the shuttle walk, and the 6-min walk. Although often not performed in a standardized manner, stair climbing can predict .VO(2)max. In general terms, patients who can climb five flights of stairs have O(2)max > 20 mL/kg/min. Conversely, patients who cannot climb one flight of stairs have .VO(2)max < 10 mL/kg/min. Data on the shuttle walk and 6 min walk are limited, but patients who cannot complete 25 shuttles on two occasions will have .VO(2)max < 10 mL/kg/min. Desaturation during an exercise test has been associated with an increased risk for perioperative complications. Lung volume reduction surgery (LVRS) for patients with severe emphysema is a controversial procedure. Some reports document substantial improvements in lung function, exercise capability, and quality of life in highly selected patients with emphysema following LVRS. Case series of patients referred for LVRS indicate that perhaps 3 to 6% of these patients may have coexisting lung cancer. Anecdotal experience from these case series suggest that patients with extremely poor lung function can tolerate combined LVRS and resection of the lung cancer with an acceptable mortality rate and good postoperative outcomes. Combining LVRS and lung cancer resection should probably be limited to those patients with heterogeneous emphysema, particularly emphysema limited to the lobe containing the tumor. PMID- 12527571 TI - Performance characteristics of different modalities for diagnosis of suspected lung cancer: summary of published evidence. AB - STUDY OBJECTIVES: To determine the test performance characteristics of various modalities for the diagnosis of suspected lung cancer. DESIGN, SETTING, AND PARTICIPANTS: A systematic search of MEDLINE, HealthStar, and Cochrane Library databases to July 2001 and print bibliographies was performed to identify studies comparing the results of sputum cytology, bronchoscopy, transthoracic needle aspirate (TTNA), or biopsy with histologic reference standard diagnoses among at least 50 patients with suspected lung cancer. MEASUREMENT AND RESULTS: For sputum cytology, the pooled specificity was 0.99 and the pooled sensitivity was 0.66, but sensitivity was higher for central lesions than for peripheral lesions (0.71 vs 0.49, respectively). Studies on bronchoscopic procedures provided data only on diagnostic yield (sensitivity). The diagnosis of endobronchial disease by bronchoscopy in 30 studies showed the highest sensitivity for endobronchial biopsy (0.74), followed by cytobrushing (0.59) and washing (0.48). The sensitivity for all modalities combined was 0.88. Thirty studies reported on peripheral lesions. Cytobrushing demonstrated the highest sensitivity (0.52), followed by transbronchial biopsy (0.46) and BAL/washing (0.43). The overall sensitivity for all modalities was 0.69. Peripheral lesions < 2 cm or > 2 cm in diameter showed sensitivities of 0.33 and 0.62, respectively. Updating a previous meta-analysis with 19 studies revealed a pooled sensitivity of 0.90 for TTNA. A trend toward lower sensitivity was noted for lesions that were < 2 cm in diameter. The accuracy in differentiating between small cell and non-small cell cytology for the various diagnostic modalities was 0.98, with individual studies ranging from 0.94 to 1.0. The average false-positive and false-negative rates were 0.09 and 0.02, respectively. CONCLUSIONS: The sensitivity of bronchoscopy is high for endobronchial disease and poor for peripheral lesions that are < 2 cm in diameter. The sensitivity of TTNA is excellent for malignant disease. The distinction between small cell lung cancer and non-small cell lung cancer by cytology appears to be accurate. PMID- 12527572 TI - Diagnosis of lung cancer: the guidelines. AB - Lung cancer is usually suspected in individuals who have abnormal chest radiograph findings or have symptoms caused by either local or systemic effects of the tumor. The method of diagnosis of suspected lung cancer depends on the type of lung cancer (ie, small cell lung cancer or non-small cell lung cancer), the size and location of the primary tumor, the presence of metastasis, and the overall clinical status of the patient. Achieving a diagnosis and staging are usually done in concert because the most efficient way to make a diagnosis often is dictated by the stage of the cancer. The best sequence of studies and interventions in a particular patient involves careful judgment of the probable reliability of a number of presumptive diagnostic issues, so as to maximize the sensitivity and to avoid performing multiple or unnecessary invasive procedures. In this article, we consider all manner of clinical presentations of lung cancer in light of currently available diagnostic procedures. Published data supporting a particular diagnostic approach is weighed based on the quality of the benefit as well as the estimated net benefit. Recommendations are graded in terms of strength to provide clinicians with guidance as to the most efficient and approach to the diagnosis of lung cancer in individual patients. PMID- 12527573 TI - Noninvasive staging of non-small cell lung cancer: a review of the current evidence. AB - STUDY OBJECTIVES: To determine the test performance characteristics of CT scanning, positron emission tomography (PET) scanning, MRI, and endoscopic ultrasound (EUS) for staging the mediastinum, and to evaluate the accuracy of the clinical evaluation (ie, symptoms, physical findings, or routine blood test results) for predicting metastatic disease in patients in whom non-small cell lung cancer or small cell lung cancer is diagnosed. DESIGN, SETTING, AND PARTICIPANTS: Systematic searches of MEDLINE, HealthStar, and Cochrane Library databases to July 2001, and of print bibliographies. Studies evaluating the staging results of CT scanning, PET scanning, MRI, or EUS, with either tissue histologic confirmation or long-term clinical follow-up, were included. The performance of the clinical evaluation was compared against the results of brain and abdominal CT scans and radionuclide bone scans. MEASUREMENT AND RESULTS: Pooled sensitivities and specificities for staging the mediastinum were as follows: for CT scanning: sensitivity, 0.57 (95% confidence interval [CI], 0.49 to 0.66); specificity, 0.82 (95% CI, 0.77 to 0.86); for PET scanning: sensitivity, 0.84 (95% CI, 0.78 to 0.89); specificity, 0.89 (95% CI, 0.83 to 0.93); and for EUS: sensitivity, 0.78 (95% CI, 0.61 to 0.89); specificity, 0.71 (95% CI, 0.56 to 0.82). For the evaluation of brain metastases, the summary estimate of the negative predictive value (NPV) of the clinical neurologic evaluation was 0.94 (95% CI, 0.91 to 0.96). For detecting adrenal and/or liver metastases, the summary NPV of the clinical evaluation was 0.95 (95% CI, 0.93 to 0.96), and for detecting bone metastases, it was 0.90 (95% CI, 0.86 to 0.93). CONCLUSIONS: PET scanning is more accurate than CT scanning or EUS for detecting mediastinal metastases. The NPVs of the clinical evaluations for brain, abdominal, and bone metastases are > or = 90%, suggesting that routinely imaging asymptomatic lung cancer patients may not be necessary. However, more definitive prospective studies that better define the patient population and improved reference standards are necessary to more accurately assess the true NPV of the clinical evaluation. PMID- 12527574 TI - The noninvasive staging of non-small cell lung cancer: the guidelines. AB - Correctly staging lung cancer is extremely important because the treatment options and the prognosis differ significantly by stage. Several noninvasive imaging studies are available to aid in identifying disease both within and outside of the chest. Chest CT scanning is useful in providing anatomic detail that better identifies the location of the tumor, its proximity to local structures, and whether or not lymph nodes in the mediastinum are enlarged. Unfortunately, the accuracy of chest CT scanning in differentiating benign from malignant lymph nodes in the mediastinum is unacceptably low. Whole-body positron emission tomography (PET) scanning provides functional information on tissue activity and has much better sensitivity and specificity than chest CT scanning for staging lung cancer in the mediastinum. In addition, metastatic disease can be detected by PET scan. Still, positive findings of PET scans can occur from nonmalignant etiologies (eg, infections), so that tissue sampling to confirm the suspected malignancy must be performed. The clinical evaluation tool, which is composed of a thorough history and physical examination, remains the best predictor of metastatic disease. If the findings from the clinical evaluation are negative, then imaging studies such as a CT scan of the head, a bone scan, or an abdominal CT scan are unnecessary, and the search for metastatic disease is complete. If signs, symptoms, or findings from the physical examination suggest the presence of malignancy, then sequential imaging, starting with the most appropriate study based on the clues obtained by the clinical evaluation, should be performed. Abnormalities detected by all of the aforementioned imaging studies are not always cancer. Unless overwhelming evidence of metastatic disease is present on an imaging study, in situations in which it will make a difference in treatment, all abnormal scan findings require tissue confirmation of malignancy so that patients are not precluded from having potentially curative surgery. PMID- 12527575 TI - Invasive staging of non-small cell lung cancer: a review of the current evidence. AB - STUDY OBJECTIVES: To determine the test performance characteristics of transbronchial needle aspiration (TBNA), transthoracic needle aspiration (TTNA), endoscopic ultrasound-guided needle aspiration (EUS-NA), and mediastinoscopy in staging non-small cell lung cancer (NSCLC). DESIGN, SETTING, AND PARTICIPANTS: Systematic search of MEDLINE, HealthStar, and Cochrane Library databases to July 2001 and print bibliographies. Included were studies comparing staging results of TBNA, TTNA, EUS-NA, or mediastinoscopy against either tissue histologic confirmation or long-term clinical follow-up (> or = 1 year). Patients included were those with NSCLC or small cell lung cancer. MEASUREMENT AND RESULTS: For patients with lung cancer, the pooled sensitivity for TBNA was 0.76, the pooled specificity was 0.96, and the negative predictive value (NPV) was 0.71. For TTNA, the pooled sensitivity was 0.91, with an NPV of 0.78. EUS-NA had a pooled sensitivity of 0.88, a pooled specificity of 0.91, and an NPV of 0.77. For standard cervical mediastinoscopy, the pooled sensitivity was 0.81, with an NPV of 0.91. The addition of either extended cervical mediastinoscopy or anterior mediastinotomy to standard cervical mediastinoscopy appeared to improve the sensitivity of any of the procedures alone. CONCLUSIONS: Invasive clinical staging of NSCLC can be performed effectively by TBNA, TTNA, EUS-NA, or mediastinoscopy. Selection of the appropriate study is dependent on the degree of suspicion for metastatic disease, the patient's comorbid illnesses, and the availability and performance characteristics of procedural options. PMID- 12527576 TI - Lung cancer. Invasive staging: the guidelines. AB - A variety of invasive staging tests are available, including mediastinoscopy, thoracoscopy (video-assisted thoracoscopic surgery), transbronchial needle aspiration (TBNA), transthoracic needle aspiration (TTNA), and endoscopic ultrasound with fine needle aspiration (EUS-NA). Each of these tests requires specific skills, has particular risks, and has technical considerations making it more or less suitable for masses in particular locations. Therefore, direct comparisons among the tests are not possible, and the issue is to define which procedure is most useful for a particular situation. Invasive staging procedures are sometimes used to confirm the stage of a lung cancer, ie, when radiographic staging is not reliable. However, invasive staging procedures are also often used to confirm the diagnosis (ie, when the radiographic stage is reliable). The first situation requires a test with a low false-negative rate; the latter requires a test with high sensitivity. Clinicians must be clear about the question at hand and how to assess the value of a test when selecting an invasive staging procedure. When confirmation of the diagnosis is the primary issue, TBNA (or EUS NA, if available) are good choices because of high sensitivity and low morbidity. When the primary issue is to confirm that there is no involvement of mediastinal lymph nodes, mediastinoscopy appears to be best suited to most situations. When the primary goal is to confirm malignant involvement of mediastinal nodes, mediastinoscopy also appears to be best in general, although TBNA, TTNA, and EUS NA may be reasonable alternatives in certain situations. However, selection of a test will also depend on the local availability of expertise, and patient specific anatomic and physiologic considerations. Selection of the optimal approach is best achieved through a multidisciplinary discussion so that all aspects can be weighed appropriately. PMID- 12527577 TI - Treatment of early stage non-small cell lung cancer. AB - Photodynamic therapy (PDT), brachytherapy, electrocautery, cryotherapy, and Nd YAG laser therapy are therapeutic options available for management of endobronchial malignancies. All of these treatment modalities have been used for both palliation of late obstructing cancers, and more recently have been used as primary treatment of early radiographically occult cancers. We reviewed the evidence for the use of these treatment options in the management of early lung cancer. PMID- 12527578 TI - Treatment of stage I non-small cell lung carcinoma. AB - The American Joint Committee on Cancer defines stage I non-small cell lung carcinoma (NSCLC) as consisting of patients with a T1 or T2 primary tumor designation and no evidence of hilar or mediastinal nodal disease (N0) or metastatic spread (M0). Medically fit patients in this clinical stage category based on conventional staging techniques should be considered for aggressive local therapy, and curative treatment is possible. Surgical resection is the accepted treatment for patients with this stage grouping, and full lobar or greater (lobectomy, pneumonectomy) rather than sublobar (wedge resection, segmentectomy) resection is strongly suggested. There is insufficient data to suggest that one method of resection (open thoracotomy, minimally invasive techniques) is superior to another. The performance of a systematic sampling or full mediastinal lymph node dissection may improve pathologic staging but is unproven therapeutically. There are no data supporting the routine use of chemotherapy in an adjuvant or neoadjuvant setting; however, recent phase II data suggest that neoadjuvant chemotherapy is feasible and safe, and larger phase III trials are now evaluating this modality. Primary radiation therapy should be considered for inoperable patients. The use of neoadjuvant or adjuvant radiation therapy in patients with stage I NSCLC is of unproven benefit. PMID- 12527579 TI - Treatment of stage II non-small cell lung cancer. AB - Based on clinical assessment alone, patients with stage II non-small cell lung cancer (NSCLC) comprise only 5% of all patients with NSCLC. In addition, patients with stage II NSCLC represent a heterogeneous group, since stage II consists of patients with T1-2N1 or T3N0 tumors. By definition, patients with tumor invading the chest wall apex, mediastinum, diaphragm, or even the mainstem bronchus may all have T3 tumors. The extent of the data available regarding treatment of each of these different groups is therefore limited. The quality of the data is limited as well, because information often comes from small series of patients. Studies of adjuvant therapy after complete resection of stage II NSCLC are an important exception to this generalization, since data from large, randomized studies of adjuvant radiation therapy, chemotherapy, or a combination of the two are available for analysis. Superior sulcus tumors are discussed elsewhere in these guidelines. PMID- 12527580 TI - Treatment of stage IIIA non-small cell lung cancer. AB - Stage IIIA non-small cell lung cancer represents a relatively heterogeneous group of patients with metastatic disease to the ipsilateral mediastinal (N2) lymph nodes and also includes T3N1 patients. Presentations of disease range from apparently resectable tumors with occult microscopic nodal metastases to unresectable, bulky multistation nodal disease. Controversy abounds as to the optimal treatment of the various stage IIIA subsets, which is fueled by a lack of meaningful, large randomized trials. Multimodality therapy of some type appears to be preferable in stage IIIA patients. PMID- 12527581 TI - Guidelines on treatment of stage IIIB non-small cell lung cancer. AB - Stage IIIB includes patients with T4, any N, M0, and any T, N3, M0. Surgery may be indicated only for carefully selected T4N0M0 patients with or without neoadjuvant chemotherapy or chemoradiotherapy. Patients with N3 lymph node involvement are not considered as surgical candidates. For patients with unresectable disease, good performance score, and minimal weight loss, treatment with combined chemotherapy and radiotherapy has resulted in better survival than treatment with radiotherapy alone. Multiple daily fractions of radiotherapy have not resulted in improved survival compared with standard fractionation once daily. Concurrent chemoradiotherapy appears to be associated with improved survival compared with sequential chemotherapy and radiotherapy. Treatment of stage IIIB due to malignant pleural effusion is addressed in the section that deals with stage IV disease. PMID- 12527582 TI - Chemotherapeutic management of stage IV non-small cell lung cancer. AB - Stage IV non-small cell lung cancer (NSCLC) denotes the presence of metastatic disease and is largely incurable using present-day therapies. Chemotherapy remains a therapeutic option in this patient population, and there are many pertinent issues surrounding its use in patients with stage IV NSCLC. Eleven questions were framed by the American College of Chest Physicians Lung Cancer Guidelines Committee, and these were addressed by a systematic search of the available literature. The issues addressed included the identification of prognostic factors in selecting patients for chemotherapy and a critical analysis of the survival benefit provided by chemotherapy. Given the development of several new chemotherapy agents over the past decade, the impact that these agents have made was addressed as well as the definition of a standard of care regarding chemotherapeutic regimens. Given the fact that chemotherapy does not represent a curative option, other issues addressed were the optimal duration of treatment as well as its impact on symptom relief and quality of life, the role of second-line therapy, and the outcomes expectations from both first-line and second-line chemotherapy. The question of what specialty delivered the chemotherapy also was addressed. Once the data were identified, a critical analysis was undertaken attempting to objectively portray the data in support of answers for each of the questions posed. We believe the data support the fact that properly selected patients benefit from chemotherapy with regard to survival and palliation in both first-line and second-line settings. It appears that in trials addressing the duration of first-line therapy, this survival and palliative benefit occurs early, and prolonged therapy is not indicated. Therapy in this setting is cost-effective, and there are several regimens that can be considered to be "standard-of-care" options. Physicians involved in the diagnosis of these patients should be aware of the potential benefits of chemotherapy, allowing them to give recommendations to patients that are based on data derived from clinical trials. In addition, this awareness will allow them to make referrals, when appropriate, to physicians who are trained in the administration of chemotherapy and the management of patients undergoing such therapy. PMID- 12527583 TI - Lung cancer. Special treatment issues. AB - This chapter of the Lung Cancer Guidelines addresses patients with particular forms of non-small cell lung cancer that require special considerations. This includes patients with Pancoast tumors, T4N0,1M0 tumors, satellite nodules in the same lobe, synchronous and metachronous multiple primary lung cancers (MPLC), and solitary metastases. For patients with a Pancoast tumor, a multimodality approach, involving chemoradiotherapy and surgical resection, appears optimal provided appropriate staging has been carried out. Patients with central T4 tumors that do not have mediastinal node involvement are uncommon. When carefully staged and selected, however, such patients appear to benefit from resection as part of the treatment as opposed to chemoradiotherapy alone. Patients with a satellite lesion in the same lobe as the primary tumor have a good prognosis and require no modification of the approach to evaluation and treatment from what would be dictated by the primary tumor alone. On the other hand, it is difficult to know how best to treat patients with a focus of the same type of cancer in a different lobe. Although MPLC do occur, the survival results after resection for either a synchronous presentation or a metachronous presentation with an interval of < 4 years between tumors are variable and generally poor, suggesting that many of these patients may have had a pulmonary metastasis rather than a second primary lung cancer. A thorough and careful evaluation of these patients is warranted to try to differentiate between patients with a metastasis and those with a second primary lung cancer, although criteria to distinguish them have not been defined. Finally, some patients with a solitary focus of metastatic disease in the brain or adrenal gland appear to benefit substantially from resection. PMID- 12527584 TI - Small cell lung cancer. AB - Among patients with lung cancers, the proportion of those with small cell lung cancer (SCLC) has decreased over the last decade. SCLC is staged as limited-stage disease and extensive-stage disease. Standard staging procedures for SCLC include CT scans of the chest and abdomen, bone scan, and CT scan or MRI of the brain. The role for positron emission tomography scanning in the staging of SCLC has yet to be defined. Limited-stage disease is treated with curative intent with chemotherapy and radiation therapy, with approximately 20% of patients achieving a cure. The median survival time for patients with limited-stage disease is approximately 18 months. Extensive-stage disease is treated primarily with chemotherapy, with a high initial response rate of 60 to 70% and a complete response rate of 20 to 30%, but with a median survival time of approximately 9 months. Patients achieving a complete remission should be offered prophylactic cranial irradiation. Currently, there is no role for maintenance treatment or bone marrow transplantation in the treatment of patients with SCLC. Relapsed or refractory SCLC has a uniformly poor prognosis. In this section, evidence-based guidelines for the staging and treatment of SCLC are outlined. PMID- 12527585 TI - Follow-up and surveillance of the lung cancer patient following curative-intent therapy. AB - The following two distinctly different issues should be taken into account when planning patient care following curative-intent therapy for lung cancer: adequate follow-up to manage complications related to the curative-intent therapy; and surveillance to detect recurrences of the primary lung cancer and/or development of a new primary lung cancer early enough to allow potentially curative retreatment. Follow-up for complications should be performed by the specialist responsible for the curative-intent therapy and should last 3 to 6 months. Recurrences of the original lung cancer will be more likely during the first 2 years after curative-intent therapy, but there will be an increased lifelong risk of approximately 1 to 2% per year of developing a metachronous, or new primary, lung cancer. A standard surveillance program for these patients is recommended based on periodic visits, with chest-imaging studies and counseling patients on symptom recognition. Whether subgroups of patients with a higher risk of developing a metachronous lung cancer (eg, those patients whose primary lung cancer was radiographically occult or central and those patients surviving for > 2 years after treatment for small cell lung cancer) should have a more intensive surveillance program is presently unclear. The surveillance program should be coordinated by a multidisciplinary tumor board and overseen by the physician who diagnosed and initiated therapy for the original lung cancer. Smoking cessation is recommended for all patients following curative-intent therapy for lung cancer. PMID- 12527586 TI - Lung cancer. Palliative care. AB - The majority of patients who acquire lung cancer will have troublesome symptoms at some time during the course of their disease. Some of the symptoms are common to many types of cancers, while others are more often encountered with lung cancer than other primary sites. The most common symptoms are pain, dyspnea, and cough. This document will address the management of these symptoms, and it will also address the palliation of specific problems that are commonly seen in lung cancer: metastases to the brain, spinal cord, and bones; hemoptysis; tracheoesophageal fistula; and obstruction of the superior vena cava. PMID- 12527587 TI - End-of-life care in patients with lung cancer. AB - Evidence-based practice guidelines for end-of-life care for patients with lung cancer have been previously available only from the British health-care system. Currently in this setting, there has been increasing concern in attaining control of the physical, psychological, social, and spiritual distress of the patient and family. This American College of Chest Physicians'-sponsored multidisciplinary panel has generated recommendations for improving quality of life after examining the English-language literature for answers to some of the most important questions in end-of-life care. Communication between the doctor, patient, and family is central to the active total care of patients with disease that is not responsive to curative treatment. The advance care directive, which has been slowly evolving and is presently limited in application and often circumstantially ineffective, better protects patient autonomy. The problem solving capability of the hospital ethics committee has been poorly utilized, often due to a lack of understanding of its composition and function. Cost considerations and a sense of futility have confused caregivers as to the potentially important role of the critical care specialist in this scenario. Symptomatic and supportive care provided in a timely and consistent fashion in the hospice environment, which treats the patient and family at home, has been increasingly used, and at this time is the best model for end-of-life care in the United States. PMID- 12527588 TI - Lung cancer. Practice organization. AB - The management of patients with suspected or known lung cancer is becoming increasingly complex. State-of-the-art care often requires input from many sources, including pulmonology, thoracic surgery, medical oncology, radiation oncology, pathology, and radiology. Valuable contributions to care also come from nursing, social work, psychology, psychiatry, pastoral care, and palliative care, among others. As a result, multidisciplinary input into care is vital. Patients with suspected lung cancer should be expeditiously evaluated and referred for management. Clear and understandable information on the diagnosis, treatment options, and possible outcomes should be provided. Treatment recommendations should be based on locally agreed-on adaptations of clinical practice guidelines. Provisions for ongoing care should be apparent to all concerned PMID- 12527589 TI - Early aggressive intervention in cystic fibrosis: is it time to redefine our "best practice" strategies? PMID- 12527590 TI - So many drugs, so little time: the future challenge of cystic fibrosis care. PMID- 12527591 TI - Altitude pulmonary edema below 8,000 feet: what are we missing? PMID- 12527592 TI - Cheyne-stokes respiration and congestive heart failure: are oxygen stores the critical factor? PMID- 12527593 TI - Entropy isn't what it used to be: applying thermodynamics to respiration in sleep. PMID- 12527594 TI - Upper airway resistance syndrome, insomnia, and functional somatic syndromes. PMID- 12527595 TI - Respiratory failure early after lung transplantation: now that we know the extent of the problem, what are the solutions? PMID- 12527596 TI - End-of-life care and eudaemonia. PMID- 12527597 TI - Diagnosis of cardiac sarcoidosis: an imperfect science, a hesitant art. PMID- 12527598 TI - Factors influencing outcomes in cystic fibrosis: a center-based analysis. AB - CONTEXT: Guidelines for managing cystic fibrosis (CF) patients have been widely circulated, but little is known about the variations in practice between sites and their association with outcomes. OBJECTIVE: To determine whether differences in lung health existed between groups of patients attending different CF care sites and to determine whether these differences are associated with differences in monitoring and intervention. DESIGN: The analysis was conducted using data from the Epidemiologic Study of Cystic Fibrosis from 1995 through 1996. SETTING: This was an observational database collecting prospective information from a large number of CF patients undergoing routine care in North America. PARTICIPANTS: Participating sites that had at least 50 CF patients who had each made at least one visit to a center during the 2-year study period were ranked on the basis of median values for FEV(1) within each of three age groups (6 to 12 years, 13 to 17 years, and >or= 18 years). INTERVENTIONS: There were no prespecified interventions in this observational study. MAIN OUTCOME MEASURES: The frequency of patient monitoring and the use of therapeutic interventions were compared between sites in the upper and lower quartiles after stratification within the site for disease severity. RESULTS: Within-site rankings tended to be consistent across the three age groups. Patients who were treated at higher ranking sites had more frequent monitoring of their clinical status, measurements of lung function, and cultures for respiratory pathogens. These patients also received more interventions, particularly IV antibiotics for pulmonary exacerbations. CONCLUSION: We found substantial differences in lung health across different CF care sites. We found that frequent monitoring and increased use of appropriate medications in the management of CF are associated with improved outcomes. PMID- 12527600 TI - GI complications after lung transplantation in patients with cystic fibrosis. AB - STUDY OBJECTIVE: Lung transplantation is now available for patients with cystic fibrosis (CF) and end-stage lung disease. While pulmonary graft function is often considered the major priority following transplantation, the nonpulmonary complications of this systemic disease also continue. We examined the GI complications in a cohort of patients who underwent transplantation. DESIGN: This was a retrospective study of all patients with CF who underwent transplantation between March 1988 and December 1998 in Toronto. Medical records were reviewed, and a short questionnaire was mailed to patients who were alive as of December 1998. RESULTS: There were 80 bilateral lung transplants performed in 75 patients. The questionnaire was distributed to 43 patients, of whom 27 patients (63%) responded. Pancreatic insufficiency requiring enzyme intake was evident in 72 of 75 patients (96%) at the time of surgery. Of three pancreatic-sufficient patients (4%), pancreatic insufficiency was diagnosed in two patients later. Biliary cirrhosis was diagnosed in three patients prior to transplantation. Distal intestinal obstruction syndrome (DIOS) was recorded for 15 patients (20%). Ten patients had a single episode, of which eight episodes occurred early in the postoperative period. Five patients had recurrent episodes. All were medically treated, except for two patients who underwent surgery. Other complications included cholecystitis (n = 3), mucocele of the appendix (n = 1), peptic ulcer disease (n = 3), and colonic carcinoma (n = 1). CONCLUSION: GI complications after lung transplantation are common in patients with CF, and attention should be paid to the risk for DIOS in the early postoperative period. Prevention and early medical treatment are important in order to avoid acute surgery. Close collaboration with the CF clinic, in order to diagnose and treat CF-related complications, is recommended. PMID- 12527599 TI - Efficiency of pulmonary administration of tobramycin solution for inhalation in cystic fibrosis using an improved drug delivery system. AB - STUDY OBJECTIVES: To determine whether tobramycin solution for inhalation (TSI) can be administered safely and more efficiently with a new-generation aerosol device, the AeroDose 5.5 RP inhaler (Aerogen; Mountain View, CA) than with the approved PARI LC PLUS nebulizer (PARI Respiratory Equipment; Monterey, CA) with Pulmo-Aide compressor (DeVilbiss Corp; Somerset, PA). Second, we wanted to ascertain which AeroDose-delivered tobramycin dose is equivalent to the standard 300-mg dose administered with the PARI LC PLUS. DESIGN: Open-label, randomized, multicenter, single-dose, three-period, four-treatment, active- control, crossover trial. SETTING: Nine US cystic fibrosis (CF) centers. PATIENTS: Fifty three patients >or= 12 years of age with a confirmed diagnosis of CF, the ability to expectorate sputum, and FEV(1) of >or= 40% of predicted. METHODS: Subjects inhaled three single doses of TSI at 1-week intervals, as follows: conventional control treatment, 300 mg via the PARI LC PLUS; and two of three experimental treatments, 30, 60, or 90 mg via the AeroDose. FEV(1) was measured before and after dosing. After each dose, sputum and serum samples were collected at various intervals for 8 h, and urine was collected for 24 h to estimate lung and systemic tobramycin delivery. RESULTS: There were no significant differences between treatments in the change in FEV(1) 30 min after dosing or in the frequency of adverse events. Sputum and serum levels of tobramycin produced by the AeroDose 90 mg dose treatment approximated those achieved with the PARI LC PLUS 300-mg dose treatment. Nebulization times using the AeroDose inhaler were < 50% those of the PARI LC PLUS. CONCLUSIONS: Compared with the standard nebulizer, the AeroDose safely achieved an approximately threefold greater efficiency in the delivery of TSI to the lungs in less than half the time. PMID- 12527601 TI - Questions on life satisfaction for adolescents and adults with cystic fibrosis: development of a disease-specific questionnaire. AB - STUDY OBJECTIVES: The development and psychometric properties of an additional disease-specific module of the Questions on Life Satisfaction for adolescents and adults with cystic fibrosis (FLZ(M)-CF) are described. The Questions on Life Satisfaction (FLZ(M)) instrument allows the respondent to rate the subjective importance and her/his satisfaction with different domains of life. METHODS: A preliminary 16-item version of the scale was employed in a study with 243 German patients with cystic fibrosis (CF) [16 to 58 years of age; FEV(1), 15 to 121% of predicted], together with the "general life satisfaction" module and the "satisfaction with health" module of the FLZ(M). Item elimination according to the principle of least common variance resulted in the final nine-item version. RESULTS: Cronbach alpha for the FLZ(M)-CF was 0.80; the split-half reliability was 0.72. Convergent validity of the scale was indicated by Pearson correlations of r = 0.75 with the generic satisfaction with health scale of the FLZ(M), r = 0.30 with FEV(1)%, and r = - 0.26 with daily time for home therapy. The scale discriminated significantly between patients with and without need for assistance in daily life and between patients with and without a partner. CONCLUSIONS: The FLZ(M)-CF is a reliable and valid instrument, and it is short enough to be used as screening instrument. The combination of generic and disease-specific scales within the FLZ(M) allows comprehensive measurement of life satisfaction for patients with CF who are > 15 years old. PMID- 12527602 TI - High-altitude pulmonary edema at moderate altitude (< 2,400 m; 7,870 feet): a series of 52 patients. AB - STUDY OBJECTIVES: To describe a group of patients who acquired pulmonary edema at a moderate altitude of 1,400 to 2,400 m. DESIGN: Observational, retrospective chart review (1992-2000) of a series of 52 consecutive patients admitted for high altitude pulmonary edema (HAPE) that occurred at 1,400 to 2,400 m. SETTING: Emergency department of a community hospital in the French Alps (altitude, 500 m). PATIENTS: Vacationing skiers who met criteria for altitude-related pulmonary edema, and in whom other causes (infectious, cardiogenic, neurogenic, and toxic) were excluded. MEASUREMENTS AND RESULTS: All patients presented with signs of pulmonary edema. Diagnoses of infectious, cardiogenic, neurogenic, or toxic edema were ruled out in each patient. All patients were hypoxemic and had radiographic signs of pulmonary edema. Virtually all patients (96%) had dyspnea, and most (77%) had moist rales. All patients were treated with supplemental oxygen (3 to 12 L/min), bed rest, moderate fluid restriction, and continuous positive airway pressure. All recovered fully and were discharged after 4 +/- 2 days (mean +/- SD). CONCLUSION: This study suggests that HAPE at moderate altitudes is more frequent than usually reported. Patients are likely to be young, vacationing men, with no history of prior disease. The disease has a favorable prognosis, and requires simple treatment and a short hospital stay. PMID- 12527603 TI - Polymorphisms of the tyrosine hydroxylase gene in subjects susceptible to high altitude pulmonary edema. AB - STUDY OBJECTIVES: A blunted hypoxic ventilatory response (HVR) has been observed in some sufferers of high-altitude pulmonary edema (HAPE), and was proposed as a potential mechanism in its pathogenesis. Tyrosine hydroxylase (TH) is a rate limiting enzyme in the carotid body responding to hypoxia to synthesize dopamine neurotransmitter to heighten ventilation. The association of constitutional susceptibility to HAPE regarding the blunted HVR aspect with polymorphisms of the TH gene was examined. DESIGN: A cross-sectional case control study. SETTING: Shinshu University Hospital, Matsumoto, Japan. PARTICIPANTS: Forty-three subjects with a history of HAPE (HAPE group) and 51 healthy climbers without a history of HAPE (control group). MEASUREMENTS: The (TCAT)n tetranucleotide microsatellite repeats within intron 1 and Met81Val variant in exon 2 of the TH gene were investigated by polymerase chain reaction following either direct sequencing or restriction fragment length polymorphism. The HVR in 21 subjects among the HAPE group was also measured. RESULTS: No significant frequency differences could be found in terms of either of the two polymorphisms between the HAPE and control groups. Meanwhile, no relationships were observed between the HVR values of HAPE subjects and the individual alleles in both polymorphisms of the TH gene. CONCLUSION: The genetic susceptibility of HAPE, specifically the blunted HVR in HAPE, is probably not associated with the mutations of the TH gene, implying that these two polymorphisms may not be a sufficient genetic marker for predicting a predisposition to the susceptibility to HAPE. PMID- 12527604 TI - Effects of nasal continuous positive airway pressure on oxygen body stores in patients with Cheyne-Stokes respiration and congestive heart failure. AB - STUDY OBJECTIVES: The mechanism(s) by which nasal continuous positive airway pressure (CPAP) is effective in the treatment of Cheyne-Stokes respiration (CSR) in patients with congestive heart failure (CHF) remains uncertain, and may involve an increase in total oxygen body stores (dampening), changes in central and peripheral controller gain, and/or improvement in cardiac function. The purpose of this study was to evaluate the effects of nasal CPAP on total oxygen stores, as measured by the rate of fall of arterial oxyhemoglobin saturation (dSaO(2)/dt), to determine if dampening may play a role in the attenuation of CSR in patients with CHF. DESIGN: Prospective controlled trial. SETTING: University hospital. PATIENTS: Nine male patients (mean +/- SD age, 59 +/- 8 years) with CHF and a mean left ventricular ejection fraction (LVEF) of 16 +/- 4%. INTERVENTIONS AND MEASUREMENTS: All patients had known CSR, as identified on a baseline polysomnographic study. Patients then underwent repeat polysomnography while receiving nasal CPAP (9 +/- 0.3 cm H(2)O). The polysomnography consisted of recording of breathing pattern, pulse oximetry, and EEG. dSaO(2)/dt was measured as the slope of a line drawn adjacent to the falling linear portion of the arterial oxygen saturation (SaO(2)) curve associated with a central apnea. All patients underwent echocardiography and right-heart catheterization within 1 month of the study to measure LVEF and cardiac hemodynamics, respectively. RESULTS: There was a significant decrease in the apnea-hypopnea index (AHI) with nasal CPAP, from 44 +/- 27 events per hour at baseline to 15 +/- 24 events per hour with nasal CPAP (p = 0.004). When compared to baseline, dSaO(2)/dt significantly decreased with nasal CPAP from 0.42 +/- 0.15% to 0.20 +/- 0.07%/s (p < 0.001). The postapneic SaO(2), when compared to baseline, significantly increased with nasal CPAP, from 87 +/- 5% to 91 +/- 4% (p < 0.05). The preapneic SaO(2) did not significantly change, from a baseline of 96 +/- 2% to 96 +/- 3% with nasal CPAP (p = 0.8). When compared to baseline, the apnea duration and heart rate did not change with nasal CPAP. While there was a significant correlation noted between baseline postapneic SaO(2) and dSaO(2)/dt (r = 0.8, p = 0.02), no correlation was seen between baseline preapneic SaO(2) and dSaO(2)/dt (r = 0.1, p = 0.7). A significant correlation was noted between baseline dSaO(2)/dt and the AHI (r = 0.7, p = 0.02). With CPAP, there was a significant correlation noted between dSaO(2)/dt and the AHI (R = 0.7, p = 0.04), but no correlation was noted between dSaO(2)/dt and postapneic SaO(2) (R = 0.1, p = 0.8). CONCLUSION: Nasal CPAP significantly decreases dSaO(2)/dt and thus increases total body oxygen stores in patients with CSR and CHF. By increasing oxygen body stores, dampening may be one of the mechanisms responsible for the attenuation of CSR seen with nasal CPAP. PMID- 12527605 TI - Changing patterns in long-term noninvasive ventilation: a 7-year prospective study in the Geneva Lake area. AB - STUDY OBJECTIVES: To describe a 7-year follow-up (1992 to 2000) of patients who were treated by home nasal positive-pressure ventilation (NPPV) for chronic hypercapnic respiratory failure. DESIGN: Prospective descriptive study. SETTING: Two university hospitals and a pulmonary rehabilitation center. PATIENTS: Two hundred eleven patients with obstructive pulmonary disorders (58 patients) or restrictive pulmonary disorders (post-tuberculosis, 23 patients; neuromuscular diseases [NM], 28 patients; post-poliomyelitis syndrome, 12 patients; kyphoscoliosis [KYPH], 19 patients; obesity-hypoventilation syndrome [OHS], 71 patients) who were treated by long-term NPPV. INTERVENTION: Annual, elective, standardized medical evaluations. MEASUREMENTS: Pulmonary function tests, arterial blood gas levels, health status, compliance, survival and probability of pursuing NPPV, and hospitalization rates. RESULTS: Patients with OHS, NM, and KYPH had the highest probability of pursuing NPPV, while patients with COPD had the lowest values. Overall, the compliance rate was high (noncompliance rate, 15%). As of 1994, COPD and OHS became the most frequent indications for NPPV, increasing regularly, while other indications remained stable. The use of pressure-cycled ventilators progressively replaced that of volume-cycled ventilators in most indications. Hospitalization rates decreased in all groups after initiating NPPV, when compared with the year before NPPV, for up to 2 years in COPD patients, and 5 years in non-COPD patients. CONCLUSION: Major changes in patient selection for NPPV occurred during the study period with a marked increase in COPD and OHS. The shift toward less expensive pressure-cycled ventilators and the decrease in hospitalizations after initiating NPPV have had positive impacts on the cost-effectiveness of NPPV in patients with chronic respiratory failure. PMID- 12527606 TI - Approximate entropy of human respiratory movement during eye-closed waking and different sleep stages. AB - STUDY OBJECTIVE: The breath-to-breath variability of respiratory parameters changes with sleep stage. This study investigates any alteration in the approximate entropy (ApEn) of respiratory movement as a gauge of complexity in respiration, by stage of consciousness, in the light of putative brain interactions. PARTICIPANTS: Eight healthy men, who were between the ages of 23 and 29 years, were investigated. MEASUREMENTS AND RESULTS: The signals of chest wall movement and EEG were recorded from 10:30 PM to 6:00 AM. After analog-to digital conversion, the ApEn of respiratory movement (3 min) and EEG (20 s) were computed. Surrogate data were tested for nonlinearity in the original time series. The most impressive reduction in the ApEn of respiratory movement was associated with stage IV sleep, when the ApEn of the EEG was also statistically significantly decreased. A statistically significant linear relation is found between the ApEn of both variables. Surrogate data indicated that respiratory movement had nonlinear properties during all stages of consciousness that were investigated. CONCLUSION: Respiratory movement and EEG signals are more regular during stage IV sleep than during other stages of consciousness. The change in complexity described by the ApEn of respiration depends in part on the ApEn of the EEG, suggesting the involvement of nonlinear dynamic processes in the coordination between brain and lungs. PMID- 12527607 TI - The symptoms and signs of upper airway resistance syndrome: a link to the functional somatic syndromes. AB - STUDY OBJECTIVES: The functional somatic syndromes are associated with a variety of symptoms/signs of uncertain etiology. We determined the prevalence of several of those symptoms/signs in patients with sleep-disordered breathing and examined the relationship between the prevalence of the symptoms/signs and the severity of sleep-disordered breathing. DESIGN: A descriptive study without intervention. SETTING: A university sleep-disorders center located in a suburban setting. PATIENTS OR PARTICIPANTS: Three groups of 25 consecutively collected patients with sleep-disordered breathing. Groups varied in their apnea hypopnea indexes (AHIs) as follows: upper airway resistance syndrome (UARS) [AHI < 10/h), mild-to moderate obstructive sleep apnea/hypopnea (OSA/H) [AHI >or= 10 to < 40/h), and moderate-to-severe OSA/H (AHI >or= 40/h). MEASUREMENTS AND RESULTS: Patients underwent comprehensive medical histories, physical examinations, and full-night polysomnography. The diagnosis of UARS included quantitative measurement of inspiratory airflow and inspiratory effort with demonstration of inspiratory flow limitation. The percentage of women among the patients with sleep-disordered breathing (p = 0.001) and the prevalence of sleep-onset insomnia (p = 0.04), headaches (p = 0.01), irritable bowel syndrome (p = 0.01), and alpha-delta sleep (p = 0.01) was correlated with decreasing severity of AHI group. CONCLUSIONS: We conclude that patients with UARS, mild-to-moderate OSA/H and moderate-to-severe OSA/H differ in their presenting symptoms/signs. The symptoms/signs of UARS closely resemble those of the functional somatic syndromes. PMID- 12527608 TI - Reference values for nocturnal home pulse oximetry during sleep in primary school children. AB - OBJECTIVE: To provide reference values for pulse oximeter saturation (SpO(2)) in primary school children, measured at home during sleep. METHODS: Recordings of SpO(2) and signal quality from 100 children were randomly selected from a larger population-based sample intended to study the prevalence of sleep-disordered breathing. Recordings were analyzed for the duration of artifact-free recording time (AFRT), minimum SpO(2) (SATmin) and median SpO(2) (SAT(50)), the SpO(2) below which the child spent 5% of AFRT (SAT(5)), and the SpO(2) below which the child spent 10% of AFRT (SAT(10)). In addition, the time in seconds with SpO(2) or= 4% per hour of AFRT (DI(4)), the number of falls in SpO(2) to 160 U/L) was documented in 13% (14 of 110 patients) in the pyrazinamide/rifampin group and in 4% (5 of 114 patients) in the isoniazid group (p = 0.03). Severe hepatotoxicity (ALT > 1,600 U/L) occurred in 2 of 43 patients (5%) receiving pyrazinamide/rifampin prior to instituting intensive monitoring. Once more intensive monitoring of liver enzymes was implemented, severe hepatotoxicity occurred in none of 67 patients. CONCLUSION: The risk of hepatitis in patients receiving pyrazinamide/rifampin for prevention of latent tuberculosis is increased threefold as compared to patients receiving isoniazid. When patients were monitored more intensively, severe hepatotoxicity did not develop, but the difference did not reach statistical significance (p = 0.15). PMID- 12527610 TI - Serologic diagnosis of tuberculosis using a simple commercial multiantigen assay. AB - SETTING: Seven primary health clinics and a pulmonary disease specialty clinic in Rio de Janeiro City, Brazil. OBJECTIVE: To evaluate a commercial immunochromatographic test kit (ICT Tuberculosis; AMRAD-ICT; Sidney, Australia) employing five recombinant Mycobacterium tuberculosis proteins for the detection of pulmonary tuberculosis (TB). DESIGN: Serology test results were compared with duplicate sputum microscopy and culture in 277 patients with symptomatic pulmonary disease (243 with pulmonary TB and 34 with nontuberculous disease). An additional 110 healthy control subjects were also tested. RESULTS: The serology test was simple and rapid to perform and detected 64.2% of smear-positive and 46.3% of smear-negative TB patients overall. HIV co-infection was present in 15.3% of TB patients, and serology was much less sensitive (overall 27.6%) in this small group, as was microscopy (13.8%). Specificity of the serology test was 100% in healthy control subjects and 85.2% in the small number of control patients with pulmonary disease, including those with prior TB. Combined with microscopy, serology detected 72.8% of TB patients. CONCLUSION: Depending on the population studied, multiantigen serologic tests for TB may be as sensitive as microscopy, but detect a different and overlapping subset of patients. The use of multiple antigens in this kit increased test sensitivity without significant loss of specificity. Bacille Calmette-Guerin vaccination and tuberculin sensitivity did not affect serology results. Estimating specificity in clinical use will require testing a much larger cohort of symptomatic patients with nontuberculous disease. The TB diagnostic performance of this group of antigens in HIV co infected individuals was poor. PMID- 12527611 TI - Reduced intracellular Mg concentrations in patients with acute asthma. AB - STUDY OBJECTIVES: To determine the intracellular and extracellular Mg concentrations in patients with acute asthma and their correlation with parameters expressing the disease severity. PATIENTS: Thirty patients with acute asthma (FEV(1), 56% predicted [SD, 14.5]), 20 patients with stable asthma (FEV(1), 97% predicted [SD, 10]), and 20 healthy subjects (FEV(1), 97% predicted [SD, 8]). METHODS: Mg concentrations in erythrocytes and plasma were measured four times: at hospital admission, after 2 days, after 5 days, and at hospital discharge. Percentage of predicted FEV(1) and peak expiratory flow rate variability were recorded simultaneously. Similar measurements were carried in all study groups. RESULTS: Mg concentrations of healthy subjects and patients with stable asthma remained unchanged in both plasma and erythrocytes. Initial Mg content in erythrocytes was significantly lower in patients with acute asthma (1.77 fmmol per cell; 95% confidence interval [CI], 1.71 to 1.83) compared to normal subjects (1.94 fmmol per cell; 95% CI, 1.82 to 2.00) and patients with stable asthma (1.92 fmmol per cell; 95% CI, 1.87 to 1.96) [p < 0.0001], and it increased significantly after the resolution of the exacerbation (from 1.77 fmmol per cell [95% CI, 1.71 to 1.83] at hospital admission to 1.90 fmmol per cell [95% CI, 1.83 to 1.98] at hospital discharge; p < 0.0001). No correlation was observed between parameters of disease severity and the initial values of Mg concentrations in erythrocytes and plasma. CONCLUSIONS: Acute asthma is associated with lower erythrocyte Mg content while plasma levels remain unchanged. This decrease in intracellular Mg content occurs regardless of the severity of the exacerbation and returns to normal values after control has been achieved. PMID- 12527612 TI - Self-reported asthma and health-related quality of life: findings from the behavioral risk factor surveillance system. AB - STUDY OBJECTIVES: Few population-based data regarding the impact of asthma on health-related quality of life in the US adult population are available. DESIGN: Cross-sectional study of participants in 50 states in the United States. SETTING: Using data from 163,773 adult respondents in the 2000 Behavioral Risk Factor Surveillance System, we examined how self-reported asthma is associated with general self-reported health and four health-related quality-of-life measures. RESULTS: Participants with self-reported current asthma reported significantly more age-adjusted physically unhealthy days (6.5 days vs 2.9 days, p < 0.001), mentally unhealthy days (5.2 days vs 3.0 days, p < 0.001), days with activity limitation (3.7 days vs 1.6 days, p < 0.001), and unhealthy physical or mental days (10.0 days vs 5.4 days, p < 0.001) in the last 30 days than participants who never had asthma. After adjusting for age, sex, race or ethnicity, educational attainment, employment status, smoking status, physical activity status, and body mass index, the odds ratios among persons with asthma compared with persons who never had asthma, were 2.41 (95% confidence interval [CI], 2.21 to 2.63) for reporting poor or fair self-rated health, 2.26 (95% CI, 2.06 to 2.49) for reporting >or= 14 days of impaired physical health during the previous 30 days, 1.55 (95% CI, 1.40 to 1.72) for reporting >or= 14 days of poor mental health during the previous 30 days, 1.96 (95% CI, 1.73 to 2.21) for reporting >or= 14 activity limitation days, and 1.99 (95% CI, 1.84 to 2.15) for reporting >or= 14 days of physically or mentally unhealthy days during the previous 30 days. Results were consistent for all age groups, for both sexes, and for all race or ethnic groups. Participants who did not currently have asthma, but had it previously, reported having more unhealthy days with all four measures than participants who never had asthma, but fewer than participants who currently had asthma. CONCLUSIONS: These results provide additional measures to evaluate and monitor the impact of asthma on the health of the US adult population. PMID- 12527613 TI - Levalbuterol compared to racemic albuterol: efficacy and outcomes in patients hospitalized with COPD or asthma. AB - STUDY OBJECTIVES: To compare clinical efficacy, patient outcomes, and medical costs in hospitalized patients treated with levalbuterol to those treated with racemic albuterol. DESIGN: Retrospective chart review. SETTING: A 180-bed community hospital. PATIENTS: Patients admitted to Halifax Regional Hospital with a diagnosis code for COPD or asthma from July 1 to December 31, 1998, and from July 1 to December 31, 1999, were eligible. In 1998, 125 patients were treated with nebulized racemic albuterol (2.5 mg q4h). In 1999, 109 patients were treated with levalbuterol (1.25 mg q8h). MEASUREMENTS AND RESULTS: Clinical efficacy was evaluated by the number of nebulizer treatments, improvement in symptoms and objective clinical findings, the length of hospital stay, and hospital discharge disposition. Medication and total hospital costs were calculated based on Red Book listings and Medicare reimbursement rates. Levalbuterol-treated patients required significantly fewer treatments with beta-agonists (mean [+/- SD] number of treatments, 19.0 +/- 12.7 vs 30.8 +/- 24.0; p < 0.001) and ipratropium bromide (mean number of treatments, 9.4 +/- 11.5 vs 23.2 +/- 25.1; p < 0.001) than did racemic albuterol-treated patients. The mean length of hospital stay in the levalbuterol group was almost 1 day less than that in the racemic albuterol group (4.7 +/- 2.9 vs 5.6 +/- 4.2 days, respectively; p < 0.058). Significantly more patients were readmitted to the hospital within 30 days in the racemic albuterol group compared with the levalbuterol group (16.4% vs 5.7%, respectively; p = 0.01). The mean total cost of nebulizer therapy was significantly greater for patients receiving racemic albuterol than for those receiving for levalbuterol ($112 +/- 101 vs $61 +/- 43, respectively; p < 0.001). The mean total hospital costs per patient were less for levalbuterol compared with racemic albuterol ($2756 +/- 2079 vs $3225 +/- 2714, respectively; p = 0.11). Regression analysis controlling for diagnosis, baseline FEV(1), and ipratropium use indicated that levalbuterol was associated with a length-of-stay savings of 0.91 days (p = 0.015), a total cost savings of $556 (p = 0.013), and a decrease in the likelihood of hospital readmission of 67% (p = 0.056). CONCLUSION: Compared with patients treated with racemic albuterol, those treated with levalbuterol required less medication, had shorter lengths of hospital stay, had decreased costs for nebulizer therapy and hospitalization, and appeared to have a more prolonged therapeutic benefit. These findings support using levalbuterol as first-line therapy for hospitalized adults with COPD or asthma. PMID- 12527614 TI - Evaluation of health-related quality of life in low-income patients with COPD receiving long-term oxygen therapy. AB - STUDY OBJECTIVE: To assess health-related quality of life (HRQL) in a low-income population of patients with hypoxemia and COPD receiving long-term oxygen therapy (LTOT). DESIGN: Cross-sectional study. SETTING: Large, tertiary care, university teaching hospital. PATIENTS OR PARTICIPANTS: Thirty-six patients with COPD requiring LTOT (mean age, 63.5 years; mean FEV(1), 32.1% of predicted; PaO(2), 50.2 mm Hg) and 33 control subjects with COPD but no severe hypoxemia (mean age, 63.1 years; FEV(1), 35.7%; PaO(2), 66.5 mm Hg). INTERVENTIONS: Patients underwent pulmonary function testing to assess physiologic function and the degree of respiratory impairment. A baseline dyspnea index (BDI) was used to determine levels of dyspnea, and a 6-min walk test was performed to evaluate physical performance and exercise capacity. The St. George Respiratory Questionnaire (SGRQ) and the Medical Outcomes Study Short-Form 36-item questionnaire (SF-36) were used to assess health status and HRQL. MEASUREMENTS AND RESULTS: The scores on the SGRQ and SF-36 indicated severe impairment. Patients receiving LTOT showed a trend toward worse scores on most dimensions of the SGRQ and SF-36, but differences between groups were only statistically significant for the physical functioning and social functioning dimensions of the SF-36. Dyspnea, as measured by the BDI, significantly correlated with all questionnaire domains except the SF 36 pain index. CONCLUSIONS: The HRQL of these low-income patients with COPD was markedly impaired, with more pronounced impairment in those receiving LTOT. The severity of dyspnea was a significant predictor of various components of quality of life in these patients. PMID- 12527615 TI - Feasibility of high-intensity, interval-based respiratory muscle training in COPD. AB - BACKGROUND: Specific respiratory muscle training can improve respiratory muscle function in patients with COPD, but the magnitude of improvement appears dependent on the magnitude of the training load. High training loads are difficult to achieve using conventional, constant loading techniques, but may be possible using interval-based training techniques. METHODS: To assess the feasibility of high-intensity respiratory muscle training, nine subjects with moderate-to-severe COPD (FEV(1) 34 +/- 12% predicted [mean +/- SD]) completed 8 weeks of interval-based respiratory muscle training combined with a general exercise program. This involved three 20-min sessions per week, each session comprising seven 2-min bouts of breathing against a constant inspiratory threshold load, each bout separated by 1 min of unloaded recovery. Inspiratory load was progressively incremented. Respiratory muscle strength (maximum inspiratory pressure generated against an occluded airway [PImax]) and endurance (maximum pressure generated against a progressively increasing inspiratory threshold load [Pthmax]) were measured before and immediately after the 8-week training period. RESULTS: By the third training session (week 1), subjects breathed against a threshold that required generation of pressures equivalent to 68 +/- 5% of the pretraining PImax. By week 8, this had increased to 95 +/- 12% of the pretraining PImax. On completion of training, PImax had increased by 32 +/ 27% (p < 0.05), Pthmax had increased by 56 +/- 33% (p < 0.05), and Pthmax/PImax had increased by 20 +/- 20% (p < 0.05). CONCLUSIONS: This study has demonstrated that high-intensity, interval-based respiratory muscle training is feasible in patients with moderate-to-severe COPD, resulting in significant improvements in respiratory muscle strength and endurance when performed three times a week for 8 weeks. PMID- 12527616 TI - Complications of tracheostomy performed in the ICU: subthyroid tracheostomy vs surgical cricothyroidotomy. AB - BACKGROUND: The morbidity of surgical tracheostomy performed in critically ill patients is not well-known. Accordingly, the aim of this prospective study was to determine the incidence and severity of complications associated with subthyroid tracheostomy and cricothyroidotomy when performed in the ICU. METHODS: Over a 2 year period, individual consecutive patients who were undergoing an elective tracheostomy were studied. Attending physicians elected the timing and technique of the tracheostomy. All procedures were performed at the bedside. A complete laryngeal examination was performed before ICU discharge, prior to decannulation, and 6 months after the tracheostomy. RESULTS: A tracheostomy (subthyroid, 86 patients; cricothyroidotomy, 32 patients) was performed in 118 of 1,574 patients (mean [+/- SD] age, 54 +/- 18 years; 79 men, 39 women; mean APACHE [acute physiology and chronic health evaluation] II score, 19 +/- 2). No deaths could be attributed to the tracheostomy procedure, and 40 complications occurred in 36 patients (30%), with a similar incidence in both groups (subthyroid group, 30 of 86 patients; cricothyroidotomy, 10 of 32 patients; p = 0.9). The severity and timing of complications were comparable between groups. CONCLUSIONS: In the present series, the incidence and severity of complications associated with conventional subthyroid tracheostomy and surgical cricothyroidotomy performed in the ICU were similar. The bedside cricothyroidotomy, which is technically easier to perform, represents a valuable alternative to conventional tracheostomy in the management of critically ill patients. PMID- 12527617 TI - Intrahospital transport of critically ill patients using ventilator with patient triggering function. AB - OBJECTIVE: To compare a new transport ventilator to manual ventilation in terms of maintaining the respiratory and hemodynamic levels of critically ill patients. DESIGN: Prospective, randomized, single-center study. SETTING: ICU in a university hospital. PATIENTS: A total of 16 patients (22 transports) who were spontaneously breathing and required ventilatory assistance on excursions from the ICU. METHODS: For each transport, the patient was randomly assigned to receive either manual ventilation (group M) or mechanical ventilation (group V). For transports in group V, the ventilators were set the same as in the ICU. Respiratory and hemodynamic variables were measured 30 min before transport (T(0)), on arrival at the site of procedure (T(1)), on return to the ICU (T(2)), and 30 min after return the ICU (T(3)). RESULTS: After transport, five patients in group M showed a significant deterioration in PaO(2)/fraction of inspired oxygen ratio, while one patient in group V showed deterioration (p = 0.056). The mean (+/- SD) respiratory rate in group M at T(2) (32 +/- 9 breaths/min) was significantly higher (p < 0.001) than at T(0) (19 +/- 6 breaths/min) and also was higher (p < 0.01) than in group V at T(2) (19 +/- 6 breaths/min). The mean tidal volume and positive end-expiratory pressure in group M at T(2) showed significantly larger variation (p < 0.05 and p < 0.001, respectively) than in group V. CONCLUSIONS: The transport ventilator that was recently approved by the US Food and Drug Administration reliably provides more stable ventilatory support than does manual ventilation. Generally, the use of this transport ventilator for intrahospital transport is preferable to manual ventilation. PMID- 12527618 TI - Respiratory failure after lung transplantation. AB - STUDY OBJECTIVES: To characterize patients who acquired postoperative respiratory failure after lung transplantation (LT), and to identify risks associated with postoperative respiratory failure and poor surgical outcome. STUDY DESIGN: Retrospective clinical analysis in a tertiary care transplantation center. METHODS: We reviewed the records of 80 consecutive patients who underwent LT from April 1994 to May 1999, analyzing their records for a number of preoperative and perioperative variables and complications. RESULTS: Forty-four patients (55%) acquired postoperative respiratory failure and had a mortality rate of 45%. No difference was noted between patients with respiratory failure and those without in terms of age (mean +/- SD, 56 +/- 9 years vs 53 +/- 11 years), gender, baseline pretransplant arterial blood gas analysis (PaCO(2), 46 +/- 9 mm Hg vs 44 +/- 10 mm Hg), and cardiopulmonary exercise testing (maximum oxygen uptake, 0.76 +/- 0.44 L/min/m(2) vs 0.82 +/- 0.20 L/min/m(2)). Ischemic reperfusion lung injury (IRLI) [55%] and perioperative cardiovascular/hemorrhagic events (36%) were the major contributors to the development of respiratory failure. Preoperative pulmonary hypertension, right ventricular (RV) dysfunction, ischemic times, and need for bilateral LT and cardiopulmonary bypass (CPB) were higher in patients with respiratory failure (p < 0.05) compared to recipients without respiratory failure. However, the presence of preoperative moderate-to-severe RV dysfunction was the only independent factor (odds ratio, 21.9; 95% confidence interval, 1.6 to 309.0). CONCLUSION: Respiratory failure after LT is common and is associated with high morbidity and mortality. Respiratory failure often occurred in patients with operative technical complications, cardiovascular events, and postoperative IRLI, which were observed most in patients requiring CPB because of RV dysfunction. PMID- 12527619 TI - Microbiological testing and outcome of patients with severe community-acquired pneumonia. AB - STUDY OBJECTIVES: The study documents the impact of microbiological investigations on therapeutic decisions and outcome in patients with severe community-acquired pneumonia (SCAP). DESIGN: Retrospective analysis of prospectively collected data. SETTING: ICUs in two teaching Spanish hospitals. PATIENTS: Two hundred four consecutive patients admitted to intensive care with SCAP. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: One hundred six patients required intubation, while 98 other patients did not (81 of these patients were managed with noninvasive mechanical ventilation). The microbiologic diagnosis was established in 57.3% of patients. The most common pathogens were Streptococcus pneumoniae, Legionella pneumophila, and Haemophilus influenzae. Pseudomonas (6.6.% vs 1.0%, p < 0.05) and Legionella (15.1% vs 7.1%, p < 0.05) were more frequently documented in intubated patients. Overall mortality was 23.5% (44.3% in intubated patients), with S pneumoniae (n = 7), Pseudomonas aeruginosa (n = 7), and L pneumophila (n = 5) being the most common lethal pathogens. Bacteriological investigation led to changes in antibiotic prescription in 41.6% of patients, including 11 patients (5%) in whom initial treatment was ineffective against the microbial isolates. The most frequent reason for changes was simplification of therapy in 65 episodes (31.8%). CONCLUSIONS: We conclude that microbiological testing is fully justified in patients with SCAP, because identifying the causative agent and adjusting treatment both impact on patient outcome. Our findings suggest that intubated patients should be empirically treated for Pseudomonas and Legionella while awaiting bacteriology results. PMID- 12527620 TI - Nuclear accumulation of p53 is a potential marker for the development of squamous cell lung cancer in smokers. AB - STUDY OBJECTIVES: To determine whether the nuclear accumulation of p53 in patients with early bronchial neoplasia represents an altered susceptibility for the development of lung cancer. PATIENTS AND MEASUREMENTS: We evaluated the percentage of cells accumulating nuclear p53 immunohistochemically in squamous cell carcinoma (SCC) of the lung, the associated uninvolved bronchial mucosa, and epithelial hyperplasia in 60 archival lung specimens of smokers and in the normal bronchial epithelium and hyperplastic lesions of 60 smokers who had not developed lung cancer. RESULTS: The percentage of cells accumulating p53 was significantly higher in SCC-associated uninvolved bronchial epithelia of (mean [+/- SD], 4 +/- 0.9%) and in specimens from patients with epithelial hyperplasia (mean, 9 +/- 2%) compared to the percentage of cells from the bronchial epithelia of (mean, 0.5 +/ 0.2%) and in specimens from patients with epithelial hyperplasia (mean, 1.5 +/- 0.5%) who were smokers who had not developed lung cancer (p = 0.0002 and p = 0.0004, respectively). We also observed a statistically significant stepwise increase in the percentage of cells accumulating p53 from SCC-associated uninvolved bronchial epithelium to those from a patient with epithelial hyperplasia to those from a patient with SCC (mean, 35 +/- 4%), suggesting the involvement of p53 accumulation in the development of SCC (p or= 357 pmol/l) identified patients with an LVEF of 40% was more than 97%. This probability rapidly decreased to 70% as the measured NT-proBNP increased to 150% of the predicted value. CONCLUSIONS: A single measurement of NT-proBNP at the time of hospital admission provides important information about LVEF in unselected patients. PMID- 12527665 TI - Comprehensive cardiac rehabilitation programme for implantable cardioverter defibrillator patients: a randomised controlled trial. AB - OBJECTIVE: To investigate the effects of a 12 week comprehensive cardiac rehabilitation (CCR) programme on patients who have undergone implantation of an implantable cardioverter-defibrillator (ICD). DESIGN: Sixteen patients with ICDs (14 (88%) male, mean (SD) age 58 (10) years, range 34-74 years) were randomised to either attend an individually tailored CCR programme or receive usual care. They then changed to the alternative regimen for a further 12 weeks. Exercise capacity was assessed using a treadmill exercise test at baseline, after usual care, after CCR and 12 weeks after CCR to assess maintenance effects. Hospital anxiety and depression (HAD) scores were recorded at each stage. RESULTS: Exercise times (min:s; mean (SD)) increased by 16% from a baseline mean of 9:55 (2:33) to 11:11 (2:17) following attendance at CCR (95% confidence interval (CI) 0:34 to 1:58; p = 0.001). This improvement was maintained 12 weeks after attendance at CCR, at 11:20 (2:17) (p = 1.00). HAD scores for anxiety and depression decreased during CCR from a baseline of 13.4 (3.6) to 8.1 (3.6), 95% CI 3.5 to 7.0 (p < 0.001) and 9.9 (3.4) to 6.7 (2.9), 95% CI 1.9 to 4.4 (p = 0.002), respectively. These improvements were maintained at 12 weeks after CCR. No ventricular arrhythmias or ICD discharges occurred during the exercise components of the CCR. The total number of ventricular arrhythmias and ICD discharges was similar 12 weeks before, during, and 12 weeks after CCR. CONCLUSIONS: CCR appears to be safe for patients with ICDs. It can improve exercising ability and lower the levels of psychological distress. A larger multicentre study is recommended to confirm these findings. PMID- 12527666 TI - Mitral annular calcification: a marker of severe coronary artery disease in patients under 65 years old. AB - BACKGROUND: Mitral annular calcification has been associated with various systemic and cardiac diseases, with a higher prevalence in women and patients over 70. A possible association between mitral annular calcification and coronary artery disease has recently been suggested. OBJECTIVE: To determine the prevalence of severe coronary artery disease in younger patients with mitral annular calcification. METHODS: Consecutive patients aged or= 70% stenosis of at least one major epicardial coronary artery. PATIENTS: 17 735 patients were screened. Of these, 6207 (35%) had mitral annular calcification and 885 (5%) were also 65 years, and chronic heart failure were considered to be clinical risk factors for thromboembolism. SETTING: Tertiary cardiac care centre. PATIENTS: 301 consecutive patients with non-rheumatic atrial fibrillation scheduled for TOE. RESULTS: 255 patients presented with clinical risk factors. 158 patients had reduced left atrial blood flow velocities, dense spontaneous echo contrast, or both. Logistic regression analysis showed that a reduced left ventricular ejection fraction and age > 65 years were the only independent predictors of a thrombogenic milieu (both p < 0.0001). The probability of having a thrombogenic milieu increased with the number of clinical risk factors present (p < 0.0001). 17.4% of the patients without clinical risk factors had a thrombogenic milieu whereas 41.2% of the patients presenting one or more clinical risk factors had none. CONCLUSION: There is a close relation between clinical risk factors and TOE markers of a thrombogenic milieu. In addition, TOE examination allows for the identification of patients with a thrombogenic milieu without clinical risk factors. PMID- 12527670 TI - Abnormal response to sodium channel blockers in patients with Brugada syndrome: augmented localised wall motion abnormalities in the right ventricular outflow tract region detected by electron beam computed tomography. AB - OBJECTIVE: To investigate the relation between the wall motion abnormalities and sodium channel abnormalities in cases of the Brugada syndrome. DESIGN: Consecutive prospective case-control study in a single hospital. SETTING: Tertiary referral centre. PATIENTS: 13 consecutive patients with Brugada syndrome and 13 age and sex matched control subjects. INTERVENTIONS: Each subject underwent electron beam computed tomography (EBT) and a 12 lead ECG before and after disopyramide injection. MAIN OUTCOME MEASURES: QRS width and the magnitude of ST segment elevation in the 12 lead ECG; wall motion by EBT. RESULTS: After disopyramide, EBT revealed deterioration of focal wall motion abnormalities in the right ventricular outflow tract region in eight of the 13 patients (62%). Prolongation of the QRS width after disopyramide injection in lead V2, which usually reflects the electrical activity in right ventricular outflow tract region, was greater in these eight patients (p < 0.01) than in the other five patients, in whom wall motion did not change after disopyramide. The degree of augmentation of ST segment elevation did not differ significantly between the two groups CONCLUSIONS: The deterioration of wall motion abnormalities in the right ventricular outflow tract region after disopyramide suggests the presence of functional abnormalities of the sodium channel. Some patients with Brugada syndrome may have arrhythmogenic substrates with abnormal responses to sodium channel blockers. PMID- 12527671 TI - Rarefaction of skin capillaries in normotensive offspring of individuals with essential hypertension. AB - BACKGROUND: Rarefaction of skin capillaries in people with intermittent borderline essential hypertension suggests a primary or an early abnormality that may antedate the onset of sustained hypertension. OBJECTIVE: To compare skin capillary density in subjects with and without a family history of essential hypertension. SUBJECTS: 21 normotensive individuals, one or both of whose parents had essential hypertension (mean age 39.3 years; blood pressure 124/79 mm Hg); 21 normotensive controls with no family history of hypertension (age 46.3 years; blood pressure 124/78 mm Hg). METHODS: The skin of the dorsum of the fingers was examined by intravital capillary microscopy before and after venous congestion at 60 mm Hg for two minutes. RESULTS: By analysis of variance, both baseline and maximum skin capillary density were lower in subjects with a family history of essential hypertension than in those with no family history (baseline: 67 v 79 capillaries per field, p = 0.008; maximum: 74 v 93 capillaries per field, p < 0.0005). CONCLUSIONS: Capillary rarefaction in essential hypertension may occur before the increase in blood pressure and could, at least in part, reflect a primary rather than a secondary abnormality. PMID- 12527672 TI - Sudden cardiac death caused by hypertrophic cardiomyopathy associated with midventricular obstruction and apical aneurysm. PMID- 12527673 TI - Myocardial viability, coronary flow reserve, and in-hospital predictors of late recovery of contractility following successful primary stenting for acute myocardial infarction. AB - OBJECTIVE: To assess the relation between myocardial viability, coronary flow reserve, and recovery of myocardial contractility after stenting for acute myocardial infarction. DESIGN: Consecutive sample prospective study. SETTING: University hospital. PATIENTS: 41 patients with single vessel disease and successful primary stenting for a first acute myocardial infarction. INTERVENTIONS: (201)Tl single photon emission computed tomography, contrast ventriculography, and intracoronary Doppler performed 7 (1) days after primary stenting. MAIN OUTCOME MEASURES: Regional contractility recovery assessed by contrast ventriculography at 6 (1) months' follow up. RESULTS: On univariate analysis, contractility recovery was correlated to prereperfusion anterograde and collateral flow grades (r = 0.41, p = 0.03 and r = 0.55, p = 0.0004), viability index (r = 0.55, p = 0.04), peak creatine kinase concentrations (r = -0.55, p = 0.0005), left ventricular ejection fraction (r = 0.45, p = 0.005), end diastolic pressure (r = -0.62, p < 0.0001), end systolic volume index (r = -0.47, p = 0.01), and the extent of hypokinetic area (r = -0.48, p = 0.003), but not the coronary flow reserve. On multivariate analysis, independent predictors of late contractility recovery were prereperfusion anterograde and collateral flow grades and viability index. Relative coronary flow reserve, reflecting the culprit vessel's microvascular function, was correlated only to the extent of the infarct risk area (r = -0.45, p = 0.003). CONCLUSIONS: Independent predictors of contractility recovery between the seventh day and the sixth month after successful stenting for acute myocardial infarction are prereperfusion anterograde and collateral flows and myocardial viability. The culprit vessel's microvascular dysfunction is independent of myocardial viability and contractility and correlated to the extent of "jeopardised microvasculature". PMID- 12527674 TI - Coronary stenting versus coronary bypass surgery in patients with multiple vessel disease and significant proximal LAD stenosis: results from the ERACI II study. AB - PURPOSE: To compare percutaneous coronary intervention (PCI) using stent implantation versus coronary artery bypass graft (CABG) in patients with multiple vessel disease with involvement of the proximal left anterior descending coronary artery (LAD). METHODS: 230 patients with multiple vessel disease and severe stenosis of the proximal LAD (113 with PCI, 117 with CABG). They were a cohort of patients from the randomised ERACI (Argentine randomized trial of percutaneous transluminal coronary angioplasty versus coronary artery bypass surgery in multivessel disease) II study. RESULTS: Both groups had similar baseline characteristics. There were no significant differences in 30 day major adverse cardiac events (death, myocardial infarction, stroke, and repeat procedures) between the strategies (PCI 2.7% v CABG 7.6%, p = 0.18). There were no significant differences in survival (PCI 96.4% v CABG 95%, p = 0.98) and survival with freedom from myocardial infarction (PCI 92% v CABG 89%, p = 0.94) at 41.5 (6) months' follow up. However, freedom from new revascularisation procedures (CABG 96.6% v PCI 73%, p = 0.0002) and frequency of angina (CABG 9.4% v PCI 22%, p = 0.025) were superior in the CABG group. CONCLUSION: Patients with multivessel disease and significant disease of the proximal LAD randomly assigned in the ERACI II trial to PCI or CABG had similar survival and survival with freedom from myocardial infarction at long term follow up. Repeat revascularisation procedures were higher in the PCI group. PMID- 12527675 TI - Exercise systolic blood pressures are of questionable value in the assessment of the adult with a previous coarctation repair. AB - BACKGROUND: The role of exercise testing in the follow up of adults with a coarctation repair is unclear. Exercise induced systolic hypertension has been advocated as an indication for further investigation; however, the value of exercise testing in this role has been questioned, especially in paediatric populations. OBJECTIVE: To assess the value of resting and exercise blood pressure measurements in a cohort of adults with repaired coarctation. SETTING: Tertiary referral centre for adult congenital cardiac disease. PATIENTS: 56 patients (33 male, 23 female) with a previous coarctation repair, and 33 age matched controls. MAIN OUTCOME MEASURES: Resting and exercise blood pressures, including arm-leg systolic blood pressure gradients; standard echocardiographic measurements of left ventricular mass, aortic root diameter, and repair site gradient. RESULTS: The coarctation cohort had higher resting upper limb blood pressures than the controls (systolic: 129.7 v 120.7 mm Hg, p = 0.014; diastolic: 76.8 v 72.2 mm Hg, p = 0.02). Mean resting arm-leg systolic blood pressure gradient was also higher, at 3.6 v -2.2 mm Hg, p = 0.027. However, there were no differences between the peak exercise systolic blood pressures of the two groups. Peak exercise systolic blood pressure did not correlate with resting arm-leg blood pressure gradient (r = 0.24, p = 0.13) or with repair site gradient (r = 0.14, p = 0.39). Resting upper limb systolic blood pressure and resting arm-leg systolic blood pressure gradient were related to repair site gradient (r = 0.33, p = 0.03, and r = 0.47, p = 0.002). CONCLUSIONS: Measurements of upper limb blood pressure during exercise are of limited value in the assessment of the post repair coarctation patient. If routine exercise testing is to be advocated in this population it must be for another indication. PMID- 12527676 TI - Fontan conversion to total cavopulmonary connection and arrhythmia ablation: clinical and functional results. AB - OBJECTIVES: To assess the early results of conversion from atriopulmonary to total cavopulmonary connection in patients with failing Fontan operation. DESIGN: Early clinical and instrumental evaluation of patients undergoing conversion from atriopulmonary to total cavopulmonary connection from April 1999 to November 2000. SETTING: Tertiary referral centre for congenital heart disease. PATIENTS: 11 Fontan patients (mean (SD) age 20.9 (6.7) years) with refractory arrhythmias or ventricular dysfunction. INTERVENTIONS: Total cavopulmonary connection, intraoperative ablation, and AAIR pacemaker implantation. MAIN OUTCOME MEASURES: Holter monitoring, transoesophageal atrial stimulation, ergometric test, and myocardial scintigraphy at a mean (SD) follow up of 16.8 (5.6) months. RESULTS: One early postoperative death occurred. During follow up three patients had relapse of atrial tachycardia, controlled by medical treatment, and two were pacemaker dependent. Transoesophageal stimulation did not induce atrial tachycardia in any patient. Ergometric test showed a diminished exercise tolerance in all but one patient. Mean minute ventilation and maximum oxygen consumption were 62% and 40% of their respective predicted values. Myocardial scintigraphy showed reversal of rest or exercise dysfunction in five patients and improved systemic ventricular function in seven. Mean basal ejection fraction increased from 39.4% (95% confidence interval (CI) 32% to 46%) to 46.5% (95% CI 41.7% to 51.2%) and ejection fraction on effort from 42.3% (95% CI 33.9% to 50.7%) to 50.2% (95% CI 44.5% to 55.9%). CONCLUSIONS: Our data show that total cavopulmonary connection associated with intraoperative ablation and pacemaker implantation allows for better control of arrhythmias and improves ventricular function in the majority of patients with failing Fontan. PMID- 12527677 TI - Vasospastic angina. PMID- 12527678 TI - Experience with transcatheter closure of secundum atrial septal defects using the Amplatzer septal occluder: a single centre study in 236 consecutive patients. AB - AIM: To evaluate the safety and efficacy of transcatheter closure of secundum atrial septal defects (ASD) with the Amplatzer septal occluder. METHODS: 236 consecutive patients with a significant ASD (age 6 months to 46 years, median 5 years; body weight 6.5-79 kg, median 18 kg) were considered for transcatheter closure with the Amplatzer septal occluder; 18 patients with defects that were too large or with a deficient inferior margin were excluded from attempted transcatheter closure after initial transthoracic (4) or transoesophageal echocardiography (14). RESULTS: At cardiac catheterisation, devices were not implanted in 18 patients because the stretched diameter of the ASD was too large (4), the device was unstable (4), compromised the mitral valve (1), or obstructed the upper right pulmonary vein (1); eight patients with additional systemic or pulmonary vein anomalies (5) or a Qp:Qs less than 1.5 (3) were excluded after angiographic and haemodynamic assessment. Thus ASD closure was done successfully in 200 patients (procedure time 25-210 minutes, median 66 minutes; fluoroscopy time 2.5-60 minutes, median 12 minutes), among whom 22 had multiple ASDs (14) or a septal aneurysm (8). The diameter of the devices ranged between 6-34 mm. Severe procedure related complications (retroperitoneal bleeding, air embolism) occurred in two cases. At follow up (33 days to 4.3 years, median 2.3 years) complete closure was documented in 94%, with a trivial residual shunt in 12 patients. CONCLUSIONS: The Amplatzer septal occluder is very efficient and offered interventional ASD closure in 84.7% of our group of consecutive patients, with excellent intermediate results. PMID- 12527679 TI - Heart rate variability in children with hypertrophic cardiomyopathy. PMID- 12527680 TI - Decreased serum vascular endothelial growth factor concentrations in patients with congestive heart failure. PMID- 12527682 TI - Prolonged temporary cardiac pacing using subcutaneous tunnelled active-fixation permanent pacing leads. PMID- 12527684 TI - Enhanced endothelin-1 degradation by intravenous morphine in patients with congestive heart failure: role of neutral endopeptidase 24.11. PMID- 12527685 TI - Deletion polymorphism of the angiotensin I converting enzyme gene is a potent risk factor for coronary artery ectasia. PMID- 12527686 TI - Asymptomatic ventricular pre-excitation in children and adolescents: a 15 year follow up study. PMID- 12527687 TI - In-stent stenosis: pathology and implications for the development of drug eluting stents. PMID- 12527688 TI - Chronic cor pulmonale. PMID- 12527689 TI - Clinical assessment of left ventricular diastolic function. PMID- 12527691 TI - Acute thrombosis of the sinus node artery: arrhythmological implications. AB - A 53 year old woman was referred for percutaneous coronary intervention because of a recent inferior myocardial infarction. During right coronary artery stent implantation, intermittent occlusion of the coronary side branch for the sinus node occurred, associated with intermittent sinus arrest and junctional escape rhythm. This led to speculation about the potential mechanisms for sinus node dysfunction. Degenerative fibrosis of nodal tissue is actually considered the most common cause of bradyarrhythmias. Yet, in everyday practice, no particular attention is usually paid to other potential pathogenic mechanisms such as coronary artery disease. This may be particularly true for elderly patients or patients with multiple risk factors. Thus, sinus node dysfunction may be an unrecognised marker of coronary artery disease. PMID- 12527692 TI - Intrastent sonotherapy in pulmonary vein restenosis: a new treatment for a recalcitrant problem. AB - A 2 year old boy developed recurrent pulmonary vein stenosis after surgical repair of infradiaphragmatic pulmonary venous connection. He had required implantation of stents in the left and right sided pulmonary veins at 7 and 13 months of age, respectively. By 2 years of age he had undergone three catheterisation procedures and two surgical procedures to treat recurrent pulmonary vein stenosis. His right ventricular pressure was suprasystemic and catheterisation showed severe neointimal proliferation of both left and right sided stents. At this time the stents were dilated by balloon with simultaneous intrastent sonotherapy. Three months later the patient's clinical improvement was significant, his right ventricular pressure had decreased, and Doppler velocity had decreased across both left and right sided stents. PMID- 12527693 TI - Protein C deficiency manifesting as an acute myocardial infarction and ischaemic stroke. AB - Protein C deficiency is a disorder in the coagulation cascade that results in predominantly venous thromboembolism. However, recent studies have implicated this disorder as a possible contributor to arterial thrombosis, especially myocardial infarction. There are six reported cases of myocardial infarction secondary to protein C deficiency in the literature. This is the first report of myocardial infarction and ischaemic stroke in the same patient as a manifestation of protein C deficiency. The investigation of hypercoagulable state is an essential component of the investigation of young patients with myocardial infarction. PMID- 12527694 TI - Obstructive intramural coronary amyloidosis and papillary muscle rupture. AB - Mitral papillary muscle rupture is usually caused by ischaemia as a complication of myocardial infarction. In a 76 year old patient with no significant disease or major cardiovascular risk factors, papillary muscle rupture was caused by obstructive intramural coronary amyloidosis, an unusual cause. PMID- 12527695 TI - Pharmacokinetics and metabolism of the reactive oxygen scavenger alpha-phenyl-N tert-butylnitrone in the male Sprague-Dawley rat. AB - The pharmacokinetics of the spin-trap alpha-phenyl-N-tert-butylnitrone (PBN) was investigated in male Sprague-Dawley rats. Plasma concentrations after i.v. administration (10 mg/kg) declined monoexponentially with a terminal half-life of 2.01 +/- 0.35 h and total plasma clearance (CL(p)) and volume of distribution at steady state (Vd(ss)) averaged 12.37 +/- 3.82 ml/min/kg and 1.74 +/- 0.5 l/kg, respectively. The observed CL(p) was in close agreement with the blood clearance (CL(b)) value (11.5 ml/min/kg) predicted from in vitro liver microsomal incubations suggesting that PBN CL(p) in rats is predominantly due to hepatic metabolism. Peak plasma concentration (C(max)) following p.o. (20 mg/kg) and s.c. (30 mg/kg) PBN administration was 7.35 +/- 1.92 and 3.56 +/- 0.66 microg/ml, whereas the area under the concentration-time curve from 0 to infinity was 23.89 +/- 5.84 and 15.96 +/- 3.10 microg-h/ml, respectively. The mean oral bioavailability of PBN was 85.63 +/- 20.93%. Biotransformation studies indicated the P450 2C11-catalyzed hydroxylation of PBN to M1. Potential sites of hydroxylation included the benzylic carbon resulting in phenyl-N-tert butylhydroxamic acid or the phenyl ring that would afford alpha-hydroxyphenyl-N tert-butylnitrone (HOPBN). The structure of M1 was established as alpha-4 Hydroxyphenyl-N-tert-butylnitrone (4-HOPBN) on the basis of: 1) obvious LC R(t) differences between M1 and the authentic hydroxamate standard, 2) P450 catalyzed hydroxylation of [(2)H]PBN that contained a deuterium instead of a hydrogen atom on its benzylic position and which afforded [(2)H]M1, and 3) comparison of the liquid chromatography-tandem mass spectrometry properties with a synthetic 4 HOPBN standard. We speculate that 4-HOPBN is an "active" PBN metabolite that provides an additive effect to the pharmacological action of PBN in vivo. PMID- 12527696 TI - The presence of xenobiotic transporters in rat placenta. AB - Understanding the role of transporters in placental handling of xenobiotics across the maternal-fetal interface is essential to evaluate the pharmacological and toxicological potential of therapeutic agents, drugs of abuse, and other xenobiotics to which the mother is exposed during pregnancy. Therefore, the purpose of this study was to assess mRNA levels of various transporters in placenta and to compare these to levels in maternal liver and kidney, predominant organs of excretion, to determine which transporters are likely to have a role in xenobiotic transfer within the placenta. During late stage pregnancy, relative amounts of mRNA levels of 40 genes representing 11 families/group of transporters were assessed in placenta with respect to relative maternal liver and kidney mRNA levels. Members of the following transporter families were assessed: three multidrug resistance (Mdr), six multidrug resistance-associated protein (Mrp), eight organic anion-transporting polypeptide (Oatp), three organic anion transporters (Oat), five organic cation transporters (Oct), two bile acid transporters (Na(+)/taurocholate-cotransporting polypeptide and bile salt export protein), four metal (ZnT1, divalent metal transporter 1, Menkes and Wilsons), a prostaglandin, two peptide, two sterolin, and four nucleoside transporters. Of the 40 genes evaluated, 16 [Mdr1a and 1b, Mrp1 and 5, Oct3 and Octn1, Oatp3 and 12, four metal, a prostaglandin, AbcG8, equilibrative nucleoside transporter 1 (ENT1), and ENT2] were expressed in placenta at concentrations similar to or higher than in maternal liver and kidney. The abundance of these mRNA transcripts in placenta suggests a role for these transporters in placental transport of xenobiotics and supports their role in the transport of endogenous substances. PMID- 12527697 TI - In vivo metabolic fate of the xeno-estrogen 4-n-nonylphenol in Wistar rats. AB - The distribution and the metabolic fate of 4-n-nonylphenol were investigated in male and female Wistar rats dosed orally with 1 microg/kg ("low-dose") or 10 mg/kg ("high-dose") labeled 4-n-nonylphenol. Following a 4-day metabolic balance study, neither the distribution pattern nor the residual levels of 4-n nonylphenol were found to be different between groups, and no unexpected tissue specific accumulation of 4-n-nonylphenol was detected. Most of the radioactivity was eliminated in urine, and consisted of hydrophilic metabolites very likely resulting from extensive beta-oxidation of the nonyl side chain and from the conjugation of the phenol to sulfate or to glucuronic acid. Traces of ring hydroxylated nonylphenol were also characterized. Fecal excretion was mainly associated with unchanged 4-n-nonylphenol and with side chain hydroxylated 4-n nonylphenol. Experiments carried out in pregnant rats exposed to a low-dose of 4 n-nonylphenol from day 3 to day 19 of gestation demonstrated similar metabolic pathways for this xeno-estrogen. Very limited amounts, if any, of non metabolized 4-n-nonylphenol did reach fetuses. The oxidative metabolism of 4-n-nonylphenol leads to the formation of both ring-hydroxylated and side chain hydroxylated metabolites. The latter metabolic pathway may be a major metabolic pathway for branched 4-nonyl-phenols and may be a clue to understand their biological activity. PMID- 12527698 TI - In vitro metabolism of fenthion and fenthion sulfoxide by liver preparations of sea bream, goldfish, and rats. AB - The in vitro metabolism of fenthion and its sulfoxide (fenthion sulfoxide) in sea bream (Pagrus major) and goldfish (Carassius auratus) was investigated and compared with that in rats. Fenthion was oxidized to fenthion sulfoxide and the oxon derivative, but not to its sulfone, in the presence of NADPH by liver microsomes of sea bream, goldfish, and rats. These liver microsomal activities of the fish were lower than those of rats but were of the same order of magnitude. The NADPH-linked oxon- and sulfoxide-forming activities of liver microsomes of the fish and rats were inhibited by SKF 525-A, metyrapone, alpha-naphthoflavone, and carbon monoxide. The oxidizing activity to fenthion sulfoxide was also inhibited by alpha-naphthylthiourea. Several cytochrome P450 isoforms and flavin containing monooxygenase 1 exhibited these oxidase activities. Fenthion sulfoxide was reduced to fenthion with liver cytosol of the fish and rats upon addition of 2-hydroxypyrimidine, N(1)-methylnicotinamide, or butyraldehyde, each of which is an electron donor of aldehyde oxidase, under anaerobic conditions. The activity was inhibited by menadione, beta-estradiol, and chlorpromazine, which are inhibitors of aldehyde oxidase. The activities in the fish livers were similar to those of rat liver. Aldehyde oxidase purified from the livers of sea bream and rats exhibited the reducing activity. Thus, fenthion and fenthion sulfoxide are interconvertible in fish and rats through the activities of cytochrome P450, flavin-containing monooxygenase, and aldehyde oxidase. PMID- 12527699 TI - Identification of novel variants of the flavin-containing monooxygenase gene family in African Americans. AB - Sequence polymorphisms in enzymes involved in drug metabolism have been widely implicated in the differences observed in the sensitivity to various xenobiotics. The flavin-containing monooxygenase (FMO) gene family in humans catalyzes the monooxygenation of numerous N-, P- and S-containing drugs, pesticides, and environmental toxicants. Six genes (FMO1-6) have been identified so far, but the major alleles of FMO2 and FMO6 encode nonfunctional proteins due to a nonsense mutation and splice-site abnormalities, respectively. Data on structural variants exist for human FMO2 and 3, whereas very little is known about the other FMO genes. FMO1-6 were scanned in 50 individuals of African-American descent using the method, detection of virtually all mutations-single-strand conformational polymorphism. A total of 49 sequence variants were identified in a total 1.35 megabases of scanned sequence, of which 29 were variants affecting protein structure or expression. Some of these are expected to affect the activity of the protein, including a nonsense mutation in FMO1 (R502X) and missense mutations in FMO1 (I303T), FMO4 (E339Q), and FMO5 (P457L) that occur in highly conserved amino acids. Additional deleterious substitutions in FMO2 (del337G) and FMO6 (Q105X) were also identified. Multiple structural variants in the FMO gene family were observed in this African-American sample. Some of the substitutions identified in this study might be useful markers in future association studies assessing sensitivity to environmental toxicants and common disease. PMID- 12527700 TI - Pharmacokinetics and pharmacodynamics of intravenous azosemide in mutant Nagase analbuminemic rats. AB - This paper reports 1) the increase in expression of CYP1A2 in mutant Nagase analbuminemic rats (NARs), 2) the role of globulin binding of azosemide in circulating blood in its urinary excretion and hence its diuretic effects in NARs, and 3) the significantly faster renal (CL(R)) and nonrenal (CL(NR)) clearances of azosemide in NARs. Azosemide (mainly metabolized via CYP1A2 in rats), 10 mg/kg, was intravenously administered to control rats and NARs. Northern and Western blot analyses revealed that the expression of CYP1A2 increased approximately 3.5-fold in NARs as compared with control. The plasma protein binding of azosemide in control rats and NARs was 97.9 and 84.6%, respectively. In NARs, plasma protein binding (84.6%) was due to binding to alpha (82.6%) and beta- (68.9%) globulins. In NARs, the amount of unchanged azosemide excreted in 8-h urine was significantly greater (37.7 versus 21.0% of intravenous dose) than that in control rats due to an increase in intrinsic renal active secretion of azosemide. Accordingly, the 8-h urine output was significantly greater in NARs. The area under the plasma concentration-time curve of azosemide was significantly smaller (505 versus 2790 microg. min/ml) in NARs because of markedly faster CL(R) (7.36 versus 0.772 ml/min/kg, secondary to a significant increase in urinary excretion of azosemide and intrinsic renal active secretion). Additionally, CL(NR) was significantly faster (12.4 versus 3.05 ml/min/kg, because of approximately 3.5 fold increase in CYP1A2) in NARs compared with control. Based on in vitro hepatic microsomal studies, the intrinsic M1 [a metabolite of azosemide; 5-(2-amino-4-chloro-5-sulfamoylphenyl)-tetrazole] formation clearance was significantly faster (67.0% increase) in NARs than that in control rats, and this supports significantly faster CL(NR) in NARs. Renal sensitivity to azosemide was significantly greater in NARs than in control rats with respect to 8-h urine output (385 versus 221 ml/kg) and 8-h urinary excretions of sodium, potassium, and chloride. This study supports that in NARs, binding of azosemide to alpha- and beta-globulins in circulating blood play an important role in its diuretic effects. PMID- 12527701 TI - Comparison of the formation of N-alkylprotoporphyrin IX after interaction of porphyrinogenic xenobiotics with single cDNA-expressed human P450 enzymes in microsomes prepared from baculovirus-infected insect cells and human lymphoblastoid cell lines. AB - In a previous study using microsomes from human lymphoblastoid cell lines (HLCL) containing single cDNA-expressed human cytochrome P450 (P450) enzymes, human P450 enzymes were identified that are susceptible to mechanism-based inactivation by the porphyrinogenic xenobiotics, 3-[(arylthio)ethyl]sydnone (TTMS), 3,5 diethoxycarbonyl-1,4-dihydro-2,6-dimethyl-4-ethylpyridine (4-ethylDDC) and allylisopropylacetamide (AIA). In this study, we tested the hypothesis that N alkylprotoporphyrin IX (N-alkylPP) formation following interaction of porphyrinogenic xenobiotics with single cDNA-expressed human P450 enzymes in microsomes from HLCL would occur only with P450 enzymes that had undergone mechanism-based inactivation. In a previous study, when 4-ethylDDC and NADPH interacted with human liver microsomes possessing elevated levels of CYP1A2 and 2C9, N-ethylprotoporphyrin IX (N-ethylPP) was not formed despite the fact that it was formed in microsomes from baculovirus-infected insect cell lines (BIICL) containing either CYP1A2 or 2C9. In this study, we tested the hypothesis that 4 ethylDDC underwent biotransformation by CYP3A4 present in human liver microsomes, diverting the xenobiotic from CYP1A2 and 2C9. Fluorometry was used to measure N alkylPP formation following interaction of porphyrinogenic xenobiotics and NADPH with cDNA-expressed human P450 enzymes in microsomes from HLCL or BIICL. With TTMS and 4-ethylDDC but not with AIA, N-alkylPP formation was observed only with human P450 enzymes CYP2D6, 1A2, 3A4, or 2C9 in microsomes from HLCL, which had undergone mechanism-based inactivation. Microsomes from BIICL containing CYP3A4 were added to a mixture of NADPH, 4-ethylDDC, and microsomes from BIICL containing CYP1A2 and 2C9. The addition of CYP3A4 to CYP1A2 and 2C9 did not decrease N-ethylPP formation, providing no support for the hypothesis. PMID- 12527702 TI - Metabolites of hexamethyldisiloxane and decamethylcyclopentasiloxane in Fischer 344 rat urine--a comparison of a linear and a cyclic siloxane. AB - Hexamethyldisiloxane (MM or HMDS) and decamethylcylclopentasiloxane (D(5)) are examples of a linear and a cyclic siloxane, respectively. These volatile low molecular weight siloxanes are of significant commercial importance. To aid in the pharmacokinetic investigations, major metabolites of MM and D(5) were identified in urine collected from Fischer (F-344) rats administered [(14)C]MM and [(14)C]D(5) orally and via intravenous injection. The metabolite profiles were obtained using a high-pressure liquid chromatography (HPLC) system equipped with a radioisotope detector. The metabolite elution was carried out on a C(18) column using an acetonitrile/water mobile phase. The structural assignments were based on GC-MS analysis of the tetrahydrofuran extract of urine containing the metabolites. Some of the metabolites in the extracts were first protected with trimethylsilyl groups prior to GC-MS analysis using bis(trimethylsiloxy)trifluoroacetamide or highly purified hexamethyldisiloxane. The structures were also confirmed by comparisons with synthetic (14)C-labeled metabolite standards. The following are among the major metabolites identified in the case of MM: Me(2)Si(OH)(2), HOMe(2)SiCH(2)OH, HOCH(2)Me(2)SiOSiMe(2)CH(2)OH, HOMe(2)SiOSiMe(2)CH(2)-OH, HOCH(2)Me(2)SiOSiMe(3), and Me(3)SiOH. The metabolites of D(5) are as follows: Me(2)Si(OH)(2), MeSi(OH)(3), MeSi(OH)(2)OSi(OH)(3), MeSi(OH)(2)OSi(OH)(2)Me, MeSi(OH)(2)OSi(OH)Me(2), Me(2)Si(OH)OSi(OH)Me(2), Me(2)Si(OH)OSiMe(2)OSi(OH)Me(2), nonamethylcyclopentasiloxanol, and hydroxymethylnonamethylcyclopentasiloxane. No parent MM or D(5) was present in urine The presence of certain metabolites such as HOMe(2)SiCH(2)OH and Me(2)Si(OH)(2) in MM and D(5), respectively, clearly established the occurrence of demethylation at the silicon-methyl bonds. Metabolites of the linear siloxane are structurally different from that obtained for cyclic siloxane except for the commonly present Me(2)Si(OH)(2). Mechanistic pathways for the formation of the metabolites were proposed. PMID- 12527703 TI - A novel P450-catalyzed transformation of the 2,2,6,6-tetramethyl piperidine moiety to a 2,2-dimethyl pyrrolidine in human liver microsomes: characterization by high resolution quadrupole-time-of-flight mass spectrometry and 1H-NMR. AB - We describe herein a novel metabolic fate of the 2,2,6,6-tetramethyl-piperidine (2,2,6,6-TMPi) moiety to a ring-contracted 2,2-dimethyl pyrrolidine (2,2-DMPy) in human liver microsomal incubations. The existence of this pathway was demonstrated for three compounds (I-III) of varied structures suggesting that this may be a general biotransformation reaction for the 2,2,6,6-TMPi moiety. The 2,2-DMPy metabolites formed in incubations of the three compounds with human liver microsomes were characterized by online high performance liquid chromatography coupled to a high resolution hybrid quadrupole-time-of-flight mass spectrometer. Suggested elemental composition obtained from accurate mass measurements of the molecular ions and fragment ions of the metabolites clearly indicated the loss of a mass equivalent to C(3)H(6) from the parent 2,2,6,6-TMPi functionality. Additional accurate tandem mass spectrometry data indicated that one of the original two gem-dimethyl groups was intact in the metabolite structure. Proof of a ring-contracted 2,2-DMPy structure was obtained using (1)H NMR experiments on a metabolite purified from liver microsomal incubations, which showed only two geminal methyl groups, instead of four in the parent compound. Two-dimensional correlation spectroscopy and decoupling experiments established aliphatic protons arranged in a pyrrolidine ring pattern. The fact that the formation of 2,2-DMPy metabolites in human liver microsomes was NADPH-dependent suggested that this novel metabolic reaction was catalyzed by the cytochrome P450 (P450) enzyme(s). Immunoinhibition studies in human liver microsomal incubations using anti-P450 monoclonal antibodies and experiments with insect cell microsomes containing individually expressed recombinant human P450 isozymes indicated that multiple P450 isozymes were capable of catalyzing this novel metabolic transformation. PMID- 12527704 TI - Absorption, metabolism, and excretion of etoricoxib, a potent and selective cyclooxygenase-2 inhibitor, in healthy male volunteers. AB - [(14)C]Etoricoxib (100 microCi/dose) was administered to six healthy male subjects (i.v., 25 mg; p.o., 100 mg). Following the i.v. dose, the plasma clearance was 57 ml/min, and the harmonic mean half-life was 24.8 h. Etoricoxib accounted for the majority of the radioactivity (approximately 75%) present in plasma following both i.v. and p.o. doses. The oral dose, administered as a solution in polyethylene glycol-400, was well absorbed (absolute bioavailability of approximately 83%). Total recovery of radioactivity in the excreta was 90% (i.v.) and 80% (p.o.), with 70% (i.v.) and 60% (p.o.) excreted in urine and 20% in feces after either route of administration. Radiochromatographic analysis of the excreta revealed that etoricoxib was metabolized extensively, and only a minor fraction of the dose (<1%) was excreted unchanged. Radiochromatograms of urine and feces showed that the 6'-carboxylic acid derivative of etoricoxib was the major metabolite observed (> or =65% of the total radioactivity). 6' Hydroxymethyl-etoricoxib and etoricoxib-1'-N-oxide, as well as the O-beta-D glucuronide conjugate and the 1'-N-oxide derivative of 6'-hydroxymethyl etoricoxib, were present in the excreta also (individually, < or =10% of the total radioactivity). In healthy male subjects, therefore, etoricoxib is well absorbed, is metabolized extensively via oxidation (6'-methyl oxidation >1'-N oxidation), and the metabolites are excreted largely in the urine. PMID- 12527705 TI - Characterization of in vitro metabolites of the aryl hydrocarbon receptor ligand 6-formylindolo[3,2-b]carbazole by liquid chromatography-mass spectrometry and NMR. AB - The tryptophan photoproduct 6-formylindolo[3,2-b]carbazole (FICZ) exhibits the highest aryl hydrocarbon receptor (AhR) binding affinity reported so far. In different cells, in vitro, both extracts of UV-irradiated tryptophan and the synthesized pure compound FICZ induce a rapid and transient expression of AhR regulated genes. The transient induction suggests that the biotransformation gene battery induced by AhR activation takes part in a metabolic degradation of the ligand, whereby a low steady-state level is regained. The down-regulation of AhR regulated gene expression was previously shown to be dependent on cytochrome P450 1A1 (CYP1A1). Metabolism of FICZ generates five major metabolites, which appeared as three peaks (M1-M3) in the high performance liquid chromatography. The aim of the present study was to use rat liver S9 from Aroclor-pretreated rats to produce large enough quantities of FICZ metabolites for structure characterization and to determine their product precursor relationship. NMR analysis of large combined fractions of the metabolites indicated that M3 and M2 contained 2 isomers, respectively. By means of liquid chromatography-mass spectrometry (negative ion electrospray mode) and NMR spectroscopy (by (1)H-NMR, correlation spectroscopy, and nuclear Overhauser effect spectroscopy techniques) five metabolites of FICZ were identified, and their structures were elucidated. The molecular weights of the two M3 isomers were 300 and both M2 and M1 compounds demonstrated molecular weights of 316, corresponding to addition of one (M3) and of two oxygen (M2 and M1), respectively. The structures were assigned as 2- and 8-hydroxy (M3), 2,10- and 4,8-dihydroxy (M2) and 2,8-dihydroxy derivatives of indolo[3,2-b]carbazole-6 carboxaldehyde (6-formylindolo[3,2-b]carbazole). PMID- 12527706 TI - Morphological quantification of emphysema in small human lung specimens: comparison of methods and relation with clinical data. AB - Small human lung specimens are frequently used for cell biological studies of the pathogenesis of emphysema. In general, lung function and other clinical parameters are used to establish the presence and severity of emphysema/chronic obstructive pulmonary disease without morphological analysis of the specimens under investigation. In this study we compared three morphological methods to analyze emphysema, and evaluated whether clinical data correlate with the morphological data of individual lung samples. A total of 306 lung specimens from resected lung(lobes) from 221 patients were inflated and characterized using three morphological parameters: the Destructive Index, the Mean Linear Intercept, and Section Assessment. Morphological data were related to each other, to lung function data, and to smoking behavior. Significant correlations (P < .001) were observed between Section Assessment and Destructive Index (r = 0.92), Mean Linear Intercept with Destructive Index (r = 0.69) and Mean Linear Intercept with Section Assessment (r = 0.65). Section Assessment, being much less time consuming than Mean Linear Intercept and Destructive Index, is the parameter of choice for initial analysis. Destructive Index is the most sensitive parameter. There was a significant (P < .001), but weak correlation for all three parameters with the diffusion capacity for CO (K(CO)) (Destructive Index: r = -0.28; Mean Linear Intercept: r = -0.34; Section Assessment: r = -0.32), and with FEV(1)/IVC (Destructive Index: r = -0.29; Mean Linear Intercept: r = -0.33; Section Assessment: r = -0.28), but not with other lung function parameters. A significant difference (P < .05) between (ex-) smokers and never-smokers was observed for Destructive Index and Section Assessment. It is concluded that the application of the three morphological parameters represents a useful method to characterize emphysematous lesions in a (semi-)quantitative manner in small human lung specimens, and that Section Assessment is a suitable and fast method for initial screening. The extent of emphysema of individual lung specimens should be established by means of morphometry, rather than lung function data. PMID- 12527707 TI - VCAM (CD-106) and ICAM (CD-54) adhesion molecules distinguish keratoacanthomas from cutaneous squamous cell carcinomas. AB - Keratoacanthomas are rapidly growing benign epithelial derived neoplasms that may evolve into squamous cell carcinomas, or represent a variant of squamous cell carcinoma. ICAM (CD-54) is a ligand for the cell adhesion receptor LFA-1, shown to be important in immune stimulation that is upgraded in inflammatory cutaneous disorders. VCAM (CD-106) is an adhesion molecule normally found in stimulated endothelium, that plays a critical role in the migration of leukocytes. We examined the immunohistochemical expression of ICAM (CD-54) and VCAM (CD-106) in a series of 50 evolving, fully developed, resolving keratoacanthoma and well differentiated and poorly differentiated squamous cell carcinoma to evaluate the possible temporal and pathogenic relation of these immune recognition markers and epithelial derived tumors. ICAM (CD-54) showed an increase in expression in the fully developed keratoacanthoma and was absent in the evolving and resolved keratoacanthoma. In the squamous cell carcinomas, expression was focally observed in the well-differentiated squamous cell carcinomas with a dramatic increase seen in the poorly differentiated squamous cell carcinomas. Similarly, VCAM (CD-106) was expressed in the fully developed keratoacanthoma and was absent in the evolving and resolved keratoacanthoma. Moderate expression for VCAM (CD-106) was seen in the well-differentiated squamous cell carcinoma, and intense expression was seen in the fully developed keratoacanthoma and poorly differentiated squamous cell carcinoma. As a group, keratoacanthoma and squamous cell carcinoma are immunophenotypically distinct. There is a temporal related increase in expression of VCAM (CD-106) in conjunction with the evolution of keratoacanthoma. Increased expression of both markers is seen with squamous cell carcinoma dedifferentiation. Application of these markers might be an important adjunct in predicting the biologic behavior and pathogenesis of these lesions. PMID- 12527708 TI - HGF, MET, and matrix-related proteases in hepatocellular carcinoma, fibrolamellar variant, cirrhotic and normal liver. AB - Fibrolamellar variant is an uncommon subcategory of hepatocellular carcinoma with a better prognostic outcome. Proteinases and growth factors that are involved in the remodeling of extracellular matrix may influence the behavior of cancers. To determine whether these factors contribute to the distinct etiologies of fibrolamellar hepatocellular carcinoma and traditional hepatocellular carcinoma, we assayed hepatocyte growth factor, the hepatocyte growth factor receptor, and two hepatocyte growth factor activators, hepatocyte growth factor activator and urokinase-type plasminogen activator, in hepatocellular carcinoma, fibrolamellar hepatocellular carcinoma, cirrhotic liver and normal liver. In addition, we examined the urokinase-type plasminogen activator receptor, the type 1 plasminogen activator inhibitor, plasmin, fibrinogen, and the type IV matrix metalloproteinases. Eighteen hepatocellular carcinomas and 11 fibrolamellar hepatocellular carcinomas were obtained as paraffin embedded sections from the University of Pittsburgh Department of Pathology. Frozen tissues from a subset of cases (9 hepatocellular carcinomas, 4 fibrolamellar hepatocellular carcinomas, 12 cirrhotic livers and 2 normal livers) were also available for analysis. Antibodies against urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor, hepatocyte growth factor and hepatocyte growth factor receptor were used to analyze immunoperoxidase stained slides from the paraffin blocks. Western blot analyses using antibodies against hepatocyte growth factor, hepatocyte growth factor receptor, phosphotyrosine, hepatocyte growth factor activator, urokinase-type plasminogen activator receptor, urokinase-type plasminogen activator, plasminogen activator inhibitor-1, fibrinogen and plasmin were performed on membrane-enriched fractions from the frozen tissue, as was collagen zymography for matrix metalloproteinase-2 and matrix metalloproteinase 9. The most notable findings are as follows: hepatocyte growth factor activator was only detected in malignant tissue but not cirrhotic liver or normal liver. Although hepatocyte growth factor was detected in most samples, it was significantly elevated in 5/9 hepatocellular carcinomas. Furthermore, 8/9 fibrolamellar hepatocellular carcinomas demonstrated hepatocyte growth factor receptor levels similar to normal, whereas 8/9 hepatocellular carcinomas and 11/12 cirrhotic livers exhibited either an increase or decrease. In contrast, active matrix metalloproteinase-2, which was absent in normal liver, was elevated in fibrolamellar hepatocellular carcinoma as compared to cirrhotic liver and conventional hepatocellular carcinoma. Surprisingly, 10/12 cirrhotic livers and 2/4 fibrolamellar hepatocellular carcinomas but only 1/9 hepatocellular carcinomas were enriched for plasmin. The combined data suggest that the hepatocyte growth factor and plasmin systems tend to be operative in hepatocellular carcinoma and cirrhotic liver, more than fibrolamellar hepatocellular carcinoma. Furthermore, matrix turnover appears to be a more prominent feature of fibrolamellar hepatocellular carcinoma. These findings provide insight into the behavioral differences between hepatocellular carcinoma and the fibrolamellar variant. PMID- 12527709 TI - The role of CD10 staining in distinguishing invasive endometrial adenocarcinoma from adenocarcinoma involving adenomyosis. AB - Adenomyosis may be involved by endometrial adenocarcinoma, but in contrast to true myometrial invasion, the depth of an adenomyotic focus involved by carcinoma does not alter pathologic tumor staging. Therefore, distinction from carcinoma invading myometrium is clinically relevant. We hypothesized that CD10, a marker of non-neoplastic and neoplastic endometrial stroma, would highlight the stromal component of adenomyotic foci and be useful in this distinction. Thirty-nine cases of endometrial adenocarcinoma were analyzed and divided into three groups: I, invasive endometrial adenocarcinoma (n = 14); II, endometrial adenocarcinoma involving adenomyosis but without myometrial invasion (n = 18); and III, adenomyosis involved by endometrial adenocarcinoma with concomitant invasive component (n = 7). All cases of adenomyosis involved by endometrial adenocarcinoma demonstrated CD10 expression in the stromal cells of adenomyotic foci. Eleven of 21 cases (52%) of invasive adenocarcinoma also showed CD10 expression, at least focally, in cells immediately surrounding the infiltrating glands. Of these, two cases (from Group III) also had associated adenomyotic involvement by carcinoma. The remaining cases of invasive carcinoma were negative for CD10. Therefore, presence of CD10 staining immediately surrounding neoplastic glands does not equate with involvement of adenomyosis by endometrial adenocarcinoma. In contrast, absence of CD10 expression excludes involvement of adenomyosis by adenocarcinoma. PMID- 12527710 TI - Widespread molecular alterations present in stage I non-small cell lung carcinoma fail to predict tumor recurrence. AB - Stage I non-small cell carcinoma (NSCLC) of the lung is typically treated with surgery alone, but with a 30 to 40% recurrence rate. Prognostic factors to stratify these patients into high- and low-risk groups would be of significant clinical value, but published data are conflicting. We studied 39 Stage I NSCLC treated with resection alone, followed for a minimum of 5 years, and divided into recurrent (RC) and non-recurrent (NRC) groups (n = 12 and 27, respectively). Allelic imbalance (loss of heterozygosity, LOH) involving genomic regions containing L-myc (1p32), hOGG1 (3p26), APC/MCC (5q21), c-fms (5q33.3), p53 (17p13), and DCC (18q21), and point mutational change in K-ras-2 (12p12) were studied by PCR-based microsatellite analysis and DNA sequencing. Mutations in k ras-2 were seen in 25% and 19% of RC and NRC tumors, respectively, most frequently in adenocarcinomas. LOH in the RC and NRC respectively were 50% and 37% for L-myc, 60% and 33% for hOGG1, 60% and 50% for APC, 38% and 35% for c-fms, 78% and 75% for p53, and 17% and 45% for DCC. No statistical significance was seen comparing any of the allelic alterations with recurrence. LOH for hOGG1 and L-myc were more commonly seen in squamous cell carcinomas. Stage I NSCLC are genetically heterogeneous with respect to mutation acquisition. The approach of investigating a panel of genes for alterations can be applied to any given tumor type, and provides information on patterns of mutations/LOH that can help us better understand the molecular biology of tumorigenesis. PMID- 12527711 TI - Lytic JC virus infection in the kidneys of AIDS subjects. AB - Our objective was to investigate the role of the human polyomavirus JC virus as a possible cause of renal damage in AIDS subjects. Histology, immunohistochemistry, and molecular biology were used to evaluate the frequency of viral infection, genotypes, viral status, and the presence of rearrangements or point mutations in specific genomic regions of strains isolated from renal tissue. Formalin-fixed, paraffin-embedded sections of postmortem renal specimens obtained from 111 unselected AIDS patients were stained for routine histology and with anti-SV40 antibody. The immunohistochemically positive specimens were further investigated by means of nested polymerase chain reaction for different polyomavirus genomic regions (large T, transcriptional control region, and viral protein 1). Furthermore, the sequences of transcriptional control region and viral protein 1 were also analyzed. Immunohistochemistry was positive in seven cases (6.3%), four of which showed morphological evidence of viral replication (intranuclear inclusion bodies and/or intratubular cellular casts): in all seven cases, only epithelial tubular cells (with and without inclusion bodies) and cellular casts were stained. The JC virus genome was identified by polymerase chain reaction in five of the seven immunohistochemically positive cases; transcriptional control region and viral protein 1 were amplified in, respectively, three and four cases. Transcriptional control region sequence analysis revealed major rearrangements in all three cases, with duplications of all the transcriptional factor-binding sites, whereas no point mutations were found in the viral protein 1 region, which was characterized as Type 1A in all cases. For the first time in AIDS subjects, this study shows that although rarely, JC virus can replicate in renal tissue. Molecular biology revealed major rearrangements in the transcriptional control region that, together with other unknown factors, could justify the increased pathogenicity of this human polyomavirus. PMID- 12527712 TI - p63 expression in solid cell nests of the thyroid: further evidence for a stem cell origin. AB - Solid cell nests of the thyroid are embryonic remnants of endodermal origin that may be difficult to distinguish from squamous metaplasia, metastatic squamous carcinoma, papillary microcarcinoma, medullary carcinoma, and C-cell hyperplasia. These embryonic structures are composed of main cells and C-cells; cystic structures and mixed follicles are sometimes observed intermingled with solid cell nests. Recently, p63, a p53 homologue that is consistently expressed in basal/stem cells of stratified epithelia and plays a major role in triggering the differentiation of some specific cell lineages, has been characterized. We evaluated the immunohistochemical expression of p63, cytokeratins (CAM 5.2, AE1/AE3, 34betaE12, 7, and 20), carcinoembryonic antigen, thyroid transcription factor 1 (TTF-1), thyroglobulin, and calcitonin using the streptavidin-biotin peroxidase complex technique in 6 bona fide solid cell nests. We observed that main cells of solid cell nests are strongly decorated by p63, while C-cells and all other thyroid structures were consistently negative. Moreover, main cells expressed carcinoembryonic antigen and all cytokeratins but cytokeratin 20 and lacked TTF-1, thyroglobulin and calcitonin. In contrast to this, C-cells of solid cell nests were immunoreactive for calcitonin, CAM 5.2, AE1/AE3, and cytokeratin 7; focal immunoreactivity for TTF-1 was also observed in some C-cells. We conclude that main cells of the solid cell nests display a basal/stem cell phenotype (p63 and basal cytokeratin positivity), whereas C-cells show features of parafollicular differentiation. We conclude, furthermore, that p63 antibodies may help in distinguishing solid cell nests from their mimics. PMID- 12527713 TI - Concurrence of histologic features of steatohepatitis with other forms of chronic liver disease. AB - Steatohepatitis, of either alcoholic or nonalcoholic etiologies, is ultimately diagnosed by clinical-pathologic correlation and is characterized histologically by lesions that differ from the portal-based chronic inflammation and fibrosis of most other forms of chronic liver disease. With the increasing prevalence of steatohepatitis in our society, it is likely that some patients will have coexistent clinical and/or histopathologic findings of steatohepatitis concurrently with another form of liver disease. The aim of this study was to document clinical and histologic findings in biopsies in an academic referral center. Ninety-three non-allograft liver biopsies with lesions of both steatohepatitis and another liver disease were retrospectively identified in 85 patients. The finding of coexisting disease represented 5.5% of all hepatitis C biopsies and 4.0% of other forms of chronic liver disease in the 34 month time period. Clinical chart review of patients with concurrent disease showed the following: Group 1, patients with hepatitis C (n = 54); Group 2, patients with hepatitis C and prior or current history of more than 80 g/d alcohol consumption (n = 20); Group 3, patients with other forms of chronic liver disease (n = 11). Groups 1 and 3 had <10 g/d alcohol use. Obesity (body mass index >30) was noted in 75%, 60%, and 33% respectively, while 94%, 87% and 100% of patients were considered overweight (body mass index > or = 25). Diabetes was reported in 35%, 25%, and 9%. The concurrence of clinical and histologic features of steatohepatitis with another chronic liver disease may be a reflection of the frequency of steatohepatitis in the population at large. PMID- 12527714 TI - Germline APC mutation on the beta-catenin binding site is associated with a decreased apoptotic level in colorectal adenomas. AB - Germline mutations in APC tumor suppressor gene are responsible for familial adenomatous polyposis (FAP). A major role of these genetic changes is the constitutive activation of beta-catenin-Tcf-4 mediated transcription of nuclear target genes, but other cellular functions can be misregulated. To assess how different APC mutations can drive the early steps of colonic tumorigenesis, we studied the effect of 10 different germline-truncating alterations on the phenotype of the corresponding adenomas. A significant reduction of apoptosis, uncoupled with an increased c-myc and cyclin-D1 expression, was seen with a frameshift mutation on codon 1383, in the 20-aa repeats of the beta-catenin degradation domain, independent of a somatic alteration on the wild-type allele. The decreased apoptotic level was associated with a higher incidence of cancerization. No other APC mutation was linked with a similar effect, even in presence of a somatic allelic loss. These findings suggest that mutations in critical sites of the beta-catenin degradation domain of APC gene can convey a selective advantage to the colonic neoplastic clones by altering the apoptotic surveillance rather than enhancing the beta-catenin-Tcf-4 transcription of growth promoting genes. PMID- 12527715 TI - Expression of platelet-derived growth factor-AA is associated with tumor progression in osteosarcoma. AB - Platelet-derived growth factors are secreted by mesenchymal cells. The homodimer platelet-derived growth factor-AA especially stimulates bone cells through interaction with the platelet-derived growth factor-alpha receptor homodimer. In this study we wanted to determine the expression of the receptor and its ligand in human osteosarcomas and to correlate the expression of platelet-derived growth factor-AA and -alpha receptor with clinicopathological parameters. Fifty-seven osteosarcomas were immunohistochemically analyzed for expression of platelet derived growth factor-AA and platelet-derived growth factor-alpha receptor. Spearman's correlation coefficient revealed a strong correlation between the expression of platelet-derived growth factor-AA and platelet-derived growth factor-alpha receptor (r = 0.867). No differences were observed relative to gender, age, tumor stage, tumor location, and response to neoadjuvant chemotherapy between high or low platelet-derived growth factor-AA and platelet derived growth factor-alpha receptor expression. High platelet-derived growth factor-AA expression correlated with tumor progression in univariate analysis (P = .0415; log-rank test), whereas platelet-derived growth factor-alpha receptor expression showed a trend toward a shorter disease-free survival, which failed to reach significance (P = .0627, log-rank test). In multivariate analysis, platelet derived growth factor-AA expression remained a significant independent predictor of tumor progression (P = .021, Cox regression). Immunohistochemical analysis of platelet-derived growth factor-AA expression in osteosarcoma may be a useful marker of prognosis and may be considered as a possible target for novel therapeutic strategies. PMID- 12527716 TI - Histidine decarboxylase expression as a new sensitive and specific marker for small cell lung carcinoma. AB - Histidine decarboxylase is one of the enzymes of the amine precursor uptake and decarboxylation system and is known to be distributed in mast cells and enterochromaffin-like cells. With the hypothesis that histidine decarboxylase expression is a marker for neuroendocrine differentiation, we studied the immunoreactivity of histidine decarboxylase in neuroendocrine cells and tumors of the thyroid gland, adrenal medulla, lung, and gastrointestinal tract. Formalin fixed paraffin sections were subjected to immunohistochemistry using anti histidine decarboxylase antibody, and the sensitivity and specificity were compared with those of conventional neuroendocrine markers (CD56, chromogranin A, synaptophysin, and neuron-specific enolase). Enterochromaffin or enterochromaffin like cells, adrenal chromaffin cells, and thyroid C-cells were positive for histidine decarboxylase, and related tumors (carcinoid tumor, pheochromocytoma, medullary carcinoma) showed a high percentage of positive staining. Furthermore, we used the antibody to distinguish small cell lung carcinoma from non neuroendocrine lung carcinoma and also to detect neuroendocrine differentiation in large-cell neuroendocrine carcinoma and gastrointestinal small-cell carcinoma. The anti-histidine decarboxylase antibody stained most small cell lung carcinoma (18 of 23, sensitivity 0.78), and was rarely reactive with non-neuroendocrine lung tumors (2 of 44; specificity, 0.95). These values were close to those obtained from CD56 staining (sensitivity/specificity, 0.87/0.98). Histidine decarboxylase was also positive for 6 of 12 large cell neuroendocrine carcinomas and 4 of 7 gastrointestinal small cell carcinomas. In conclusion, we demonstrated that histidine decarboxylase is useful to distinguish between small cell lung carcinoma and non-neuroendocrine carcinoma and to demonstrate neuroendocrine differentiation. PMID- 12527717 TI - Reliability of tissue microarrays in detecting protein expression and gene amplification in breast cancer. AB - Tissue microarrays allow high throughput molecular profiling of diagnostic or predictive markers in cancer specimens and rapid validation of novel potential candidates identified from genomic and proteomic analyses in a large number of tumor samples. To validate the use of tissue microarray technology for all the main biomarkers routinely used to decide breast cancer prognostication and postsurgical adjuvant therapy, we constructed a tissue microarray from 97 breast tumors, with a single 0.6 mm core per specimen. Immunostaining of tissue microarray sections and conventional full sections of each tumor were performed using well-characterized prognostic markers (estrogen receptor ER, progesterone receptor PR and c-erbB2). The full section versus tissue microarray concordance for these stains was 97% for ER, 98% for PR, and 97% for c-erbB2, respectively, with a strong statistical association (kappa value more than 0.90). Fluorescence in situ hybridization analysis for HER-2/neu gene amplification from the single core tissue microarray was technically successful in about 90% (87/97) of the cases, with a concordance of 95% compared with parallel analyses with the full sections. The correlation with other pathological parameters was not significantly different between full-section and array-based results. It is concluded that the constructed tissue microarray with a single core per specimen ensures full biological representativeness to identify the associations between biomarkers and clinicopathological parameters, with no significant associated sampling bias. PMID- 12527718 TI - Expression of transforming growth factor-beta1 and transforming growth factor beta receptors in hepatocellular carcinoma and dysplastic nodules. AB - In this study we analyzed by immunohistochemistry the expression of TGF-beta1 protein and TGF-beta receptors I and II in 4 low-grade dysplastic nodules, 2 high grade dysplastic nodules, 6 early, 22 small, and 62 advanced hepatocellular carcinomas. The expression of TGF-beta1 protein by hepatocytes was decreased in advanced hepatocellular carcinoma compared with small or early hepatocellular carcinoma(P < .05). Frequent and intense staining of TGF-beta1 protein was noted in the sinusoidal endothelium of advanced hepatocellular carcinomas despite of its decreased staining in hepatocellular carcinoma cells. Reduced expression of TGF-beta receptors I and II compared with surrounding nontumorous tissue were noted from the early hepatocellular carcinoma stage suggesting that down regulation of TGF-beta receptors is correlated with progression from premalignant to malignant phenotype. Reduced expression of both TGF-beta1 and TGF-beta receptor II in neoplastic hepatocytes were also significantly correlated with increased tumor size and increased proliferative activity(P < .05). These findings suggest that during hepatocarcinogenesis, the inhibitory effects of TGF beta1 protein on hepatocellular carcinoma cells is outweighed by its effects on stromal elements, which, overall, contributes indirectly to a tumor growth stimulatory environment. Also, the growth-inhibitory effects of TGF-beta1 may have been further negated by reduced TGF-beta receptors on hepatocellular carcinoma cells. PMID- 12527720 TI - How to balance the brain energy budget while spending glucose differently. PMID- 12527721 TI - Metabolic and mitogenic signal transduction in human skeletal muscle after intense cycling exercise. AB - We determined whether mitogen-activated protein kinase (MAPK) and 5'-AMP activated protein kinase (AMPK) signalling cascades are activated in response to intense exercise in skeletal muscle from six highly trained cyclists (peak O(2) uptake (.V(O2,peak)) 5.14 +/- 0.1 l min(-1)) and four control subjects (Vdot;(O(2))(,peak) 3.8 +/- 0.1 l min(-1)) matched for age and body mass. Trained subjects completed eight 5 min bouts of cycling at approximately 85% of .V(O2,peak) with 60 s recovery between work bouts. Control subjects performed four 5 min work bouts commencing at the same relative, but a lower absolute intensity, with a comparable rest interval. Vastus lateralis muscle biopsies were taken at rest and immediately after exercise. Extracellular regulated kinase (ERK1/2), p38 MAPK, histone H3, AMPK and acetyl CoA-carboxylase (ACC) phosphorylation was determined by immunoblot analysis using phosphospecific antibodies. Activity of mitogen and stress-activated kinase 1 (MSK1; a substrate of ERK1/2 and p38 MAPK) and alpha(1) and alpha(2) subunits of AMPK were determined by immune complex assay. ERK1/2 and p38 MAPK phosphorylation and MSK1 activity increased (P < 0.05) after exercise 2.6-, 2.1- and 2.0-fold, respectively, in control subjects and 1.5-, 1.6- and 1.4-fold, respectively, in trained subjects. Phosphorylation of histone H3, a substrate of MSK1, increased (P < 0.05) approximately 1.8-fold in both control and trained subject. AMPKalpha(2) activity increased (P < 0.05) after exercise 4.2- and 2.3-fold in control and trained subjects, respectively, whereas AMPKalpha(1) activity was not altered. Exercise increased ACC phosphorylation (P < 0.05) 1.9- and 2.8-fold in control and trained subjects. In conclusion, intense cycling exercise in subjects with a prolonged history of endurance training increases MAPK signalling to the downstream targets MSK1 and histone H3 and isoform-specific AMPK signalling to ACC. Importantly, exercise-induced signalling responses were greater in untrained men, even at the same relative exercise intensity, suggesting muscle from previously well-trained individuals requires a greater stimulus to activate signal transduction via these pathways. PMID- 12527722 TI - The familial hemiplegic migraine mutation R192Q reduces G-protein-mediated inhibition of P/Q-type (Ca(V)2.1) calcium channels expressed in human embryonic kidney cells. AB - Familial hemiplegic migraine is associated with at least 13 different missense mutations in the alpha1A Ca(2+) channel subunit. Some of these mutations have been shown to affect the biophysical properties of alpha1A currents. To date, no study has examined the influence of such mutations on the G-protein regulation of channel function. Because G-proteins inhibit movement of the voltage sensor, we examined the effects of the R192Q mutation, which neutralizes a positive charge in the first S4 segment. Human wild-type (WT) or R192Q mutant channels were expressed in human embryonic kidney tsA-201 cells along with dopamine D2 receptors. Application of quinpirole induced fast (approximately 1 s), pertussis toxin-sensitive inhibition of alpha1A(WT) and alpha1A(R192Q) Ca(2+) currents, consistent with the activation of a membrane-delimited pathway. alpha1A(WT) Ca(2+) currents were inhibited by 62.9 +/- 0.9 % (n = 27), whereas alpha1A(R192Q) Ca(2+) currents were inhibited by only 47.9 +/- 1.8 % (n = 35; P < 0.001). Concentration-response analysis showed that only the extent of inhibition was affected, with no change in agonist potency (EC(50) = 1 nM). Prepulse facilitation, which is a characteristic of voltage-dependent inhibition, was also reduced by the R192Q mutation. However, the kinetics of facilitation and slow activation were not affected, suggesting that G-protein-Ca(2+) channel affinity was unchanged. These results show that the R192Q mutation reduces the G-protein inhibition of P/Q-type Ca(2+) channels, probably by altering mechanisms by which Gbetagamma subunit binding induces a change in channel gating. Altered G-protein modulation and the consequent reduced presynaptic inhibition may contribute to migraine attacks by favouring a persistent state of hyperexcitability. PMID- 12527723 TI - Structure, function and immunolocalization of a proton-coupled amino acid transporter (hPAT1) in the human intestinal cell line Caco-2. AB - The human orthologue of the H(+)-coupled amino acid transporter (hPAT1) was cloned from the human intestinal cell line Caco-2 and its functional characteristics evaluated in a mammalian cell heterologous expression system. The cloned hPAT1 consists of 476 amino acids and exhibits 85 % identity with rat PAT1. Among the various human tissues examined by Northern blot, PAT1 mRNA was expressed most predominantly in the intestinal tract. When expressed heterologously in mammalian cells, hPAT1 mediated the transport of alpha (methylamino)isobutyric acid (MeAIB). The cDNA-induced transport was Na(+) independent, but was energized by an inwardly directed H(+) gradient. hPAT1 interacted with glycine, L-alanine, L-proline, alpha-aminoisobutyrate (AIB) and gamma-aminobutyrate (GABA), as evidenced from direct transport measurements and from competition experiments with MeAIB as a transport substrate. hPAT1 also recognized the D-isomers of alanine and proline. With serine and cysteine, though the L-isomers did not interact with hPAT1 to any significant extent, the corresponding D-isomers were recognized as substrates. With proline and alanine, the affinity was similar for L- and D-isomers. However, with cysteine and serine, the D-isomers showed 6- to 8-fold higher affinity for hPAT1 than the corresponding L-isomers. These functional characteristics of hPAT1 closely resemble those that have been described previously for the H(+)-coupled amino acid transport system in Caco-2 cells. Furthermore, there was a high degree of correlation (r(2) = 0.93) between the relative potencies of various amino acids to inhibit the H(+)-coupled MeAIB transport measured with native Caco-2 cells and with hPAT1 in the heterologous expression system. Immunolocalization studies showed that PAT1 was expressed exclusively in the apical membrane of Caco-2 cells. These data suggest that hPAT1 is responsible for the H(+)-coupled amino acid transport expressed in the apical membrane of Caco-2 cells. PMID- 12527724 TI - Theta oscillation coupled spike latencies yield computational vigour in a mammalian sensory system. AB - Theoretical work carried out almost a decade ago proposed that subthreshold oscillations in membrane potential could be used to convert synaptic current strength into a code reliant on action potential (AP) latencies. Using whole-cell recordings we present experimental evidence for the occurrence of prominent network-driven subthreshold theta oscillations in mitral cells of the mouse olfactory bulb. Activity induced by both injected current and sensory input was accurately reflected in initial AP latency from the beginning of each oscillation cycle. In a network model we found that an AP latency code rather than AP number or instantaneous firing rate provided computational speed and high resolution, and was easily implemented. This coding strategy was also found to be invariant to the total input current as long as the relative input intensities to glomeruli remained constant. However, it was highly sensitive to changes in the ratios of the input currents and improved by lateral inhibitory mechanisms. Since the AP latency-based coding scheme was dependent on the subthreshold oscillation we conclude that the theta rhythm serves a functional role in temporally reformatting the strengths and patterns of synaptic input in this sensory system. PMID- 12527725 TI - Multidrug resistance transporters in the olfactory receptor neurons of Xenopus laevis tadpoles. AB - Olfactory receptor neurons (ORNs) are the only class of neurons that is directly exposed to the environment. Therefore, they need to deal with xenobiotic and potentially cytotoxic substances. Here we show for the first time that ORNs possess transporter systems that expel xenobiotics across the plasma membrane. Using calcein and calcium-indicator dyes as xenobiotics, we demonstrate that ORNs appear to express the multidrug resistance P-glycoprotein (MDR1) and multidrug resistance-associated proteins (MRP). This endows ORNs with the ability to transport a large number of substrates including calcium-indicator dyes and calcein across their plasma membranes. Conversely, blocking P-glycoprotein and MRP increases the net uptake of these dyes. PMID- 12527726 TI - Kv1.4 channel block by quinidine: evidence for a drug-induced allosteric effect. AB - We studied quinidine block of Kv1.4DeltaN, a K(+) channel lacking N-type inactivation, expressed in Xenopus ooctyes. Initially, quinidine intracellularly blocked the open channel so rapidly it overlapped with activation. This rapid open channel block was reduced (non-additively) by interventions that slow C-type inactivation: [K(+)](o) elevation and an extracellular lysine to tyrosine mutation (K532Y). These manipulations reduced the affinity of rapid open channel block ~10-fold, but left the effective electrical distance unchanged at ~0.15. Following rapid open channel block, there were time-dependent quinidine effects: the rate of inactivation during a single depolarisation was increased, and repetitive pulsing showed use dependence. The rate of recovery from the time dependent aspect of quinidine block was similar to recovery from normal C-type inactivation. Manipulations that prevented the channel from entering the C-type inactivated state (i.e. high [K(+)](o) or the K532Y mutation) prevented the development of the time-dependent quinidine-induced inactivation. The concentration dependence of the rapid block and the time-dependent quinidine induced inactivation were similar, but the time-dependent component was strongly voltage sensitive, with an effective electrical distance of 2. Clearly, this cannot reflect the permeation of quinidine through the electric field, but must be the result of some other voltage-sensitive change in the channel. We propose that quinidine promotes the entry of the channel into a C-type inactivated state in a time- and voltage-dependent manner. We developed a mathematical model based on these results to test the hypothesis that, following rapid open channel block, quinidine promotes development of the C-type inactivated state through a voltage dependent conformational change. PMID- 12527727 TI - Effect of androgen deficiency on mouse ventricular repolarization. AB - We previously demonstrated that female mouse ventricles have longer action potential durations (APDs) than males. This delayed repolarization results from a lower current density of the ultrarapid delayed rectifier K(+) current (I(K,ur)) and a lower expression level of its underlying K(+) channel (Kv1.5). To evaluate whether this sex difference could be attributable to the action of male sex hormones, we studied the effect of androgen deficiency on ventricular repolarization. We compared cardiac electrophysiological properties in castrated (orchiectomized; ORC) and control (CTL) male mice. Q-Tc intervals as well as APDs measured at 20 %, 50 % and 90 % of repolarization were all significantly longer in ORC than in CTL. The current density of I(K,ur) was significantly lower in ORC than in CTL (at +50 mV, ORC: 29 +/- 4 pA pF(-1), n = 25; CTL: 48 +/- 5 pA pF(-1), n = 17; P = 0.006). In contrast, all the other K(+) currents present in mouse ventricular myocytes were comparable between ORC and CTL. Moreover, results of Western blot analysis showed a lower expression level of Kv1.5 protein in ORC but no difference between the two groups for the other K(+) channels studied. This study demonstrates that androgen deficiency leads to a reduction in the density of I(K,ur) and Kv1.5 in mouse ventricle, and consequently, to prolongation of APD and Q-Tc interval. In conclusion, these findings strongly suggest that male sex hormones contribute to the sex difference that we previously reported in cardiac repolarization in adult mouse heart. PMID- 12527728 TI - Muscarinic regulation of pacemaker frequency in murine gastric interstitial cells of Cajal. AB - Peristaltic contractions in the stomach are regulated by the spread of electrical slow waves from the corpus to the pylorus. Gastric slow waves are generated and propagated by the interstitial cells of Cajal (ICC). All regions distal to the dominant pacemaker area in the corpus are capable of generating slow waves, but orderly gastric peristalsis depends upon a frequency gradient in which the corpus pacemaker frequency exceeds the antral frequency. Cholinergic, muscarinic stimulation enhances pacemaker frequency. We investigated this phenomenon using intact murine gastric muscles and cultured ICC. Acetylcholine (ACh) increased the frequency of slow waves in antrum and corpus muscles. The increase was significantly greater in the antrum. ACh and carbachol (CCh) increased the pacemaker currents in cultured ICC. At high doses of CCh, transient pacemaker currents fused into sustained inward currents that persisted for the duration of stimulation. The effects of CCh were blocked by low doses of the M(3) receptor antagonist 1-dimethyl-4-diphenylacetoxypiperidinium. Frequency enhancement by CCh was not affected by forskolin, but the phospholipase C inhibitor U-73122 inhibited both the increase in frequency and the development of tonic inward currents. 2-Aminoethyldiphenyl borate also blocked the chronotropic responses to CCh. Inhibitors of protein kinase C did not block responses to CCh. These studies show that mice are an excellent model for studying mechanisms that regulate gastric slow-wave frequency. CCh, apparently via production of inositol 1,4,5 trisphosphate, accelerates the frequency of pacemaker activity. High concentrations of CCh may block the entrainment of pacemaker currents, resulting in a tonic inward current. PMID- 12527729 TI - Differences in conductance of M2 proton channels of two influenza viruses at low and high pH. AB - The M2 protein of influenza A viruses forms a proton channel involved in modifying virion and trans Golgi pH during infection. Previous studies of the proton current using whole-cell patch clamp of mouse erythroleukaemia (MEL) cells expressing the M2 protein of the "Weybridge" strain provided evidence for two protonation sites, one involved in permeation, the other in activation by acid pH. The present report compares the M2 channels of two different strains of influenza virus, "Weybridge" (WM2) and "Rostock" (RM2). Whereas with external acid pH the current-voltage relations showed similar small degrees of inward rectification, a similar apparent K(d) of approximately 10 microM for proton permeation and a high selectivity for protons over Na(+), the two M2 proteins differed in whole-cell conductance at low and high pH. The proton conductance of unit membrane area was on average 7-fold greater in RM2- than WM2-expressing MEL cells. At high external pH WM2 was shown previously to have small conductance for outward current at positive driving potential. In contrast, RM2 shows high conductance for outward current with high external pH, but shows small conductance for inward current with high internal pH, conditions in which WM2 shows high conductance for inward current. The different properties of the conductances due to the two channels at high pH were determined by three amino acids in their transmembrane domains. All intermediate mutants possessed one or other property and transformation of the WM2 phenotype into that of RM2 required substitution in all three residues V27I, F38L and D44N; single substitutions in RM2 effected the opposite phenotypic change. The significance of this difference for virus replication is not clear and it may be that the higher proton flux associated with RM2 is the main factor determining its increased ability to dissipate pH gradients. It is apparent, however, from the specific differences in the sidedness of the pH-induced changes in voltage dependence of the whole-cell current that this is an intrinsic property of the virus proton channel which may have parallels with regulation of other proton channels. PMID- 12527730 TI - GABA B receptor modulation of excitatory and inhibitory synaptic transmission onto rat CA3 hippocampal interneurons. AB - Hippocampal stratum radiatum inhibitory interneurons receive glutamatergic excitatory innervation via the recurrent collateral fibers of CA3 pyramidal neurons and GABAergic inhibition from other interneurons. We examined both presynaptic- and postsynaptic-GABA(B) receptor-mediated responses at both synapse types. Postsynaptic GABA(B) receptor-mediated responses were absent in recordings from young (P16-18) but present in recordings from older animals (> or =P30) suggesting developmental regulation. In young animals, the GABA(B) receptor agonist, baclofen, inhibited the amplitude of evoked EPSCs and IPSCs, an effect blocked by prior application of the selective antagonist CGP55845. Baclofen enhanced the paired-pulse ratio and coefficient of variation of evoked EPSCs and IPSCs, consistent with a presynaptic mechanism of regulation. In addition, baclofen reduced the frequency of miniature IPSCs but not mEPSCs. However, baclofen reduced the frequency of KCl-induced mEPSCs; an effect blocked by Cd(2+), implicating presynaptic voltage-gated Ca(2+) channels as a target for baclofen modulation. In contrast, although Cd(2+) prevented the KCl-induced increase in mIPSC frequency, it failed to block baclofen's reduction of mIPSC frequency. Whereas N- and P/Q-types of Ca(2+) channels contributed equally to GABA(B) receptor-mediated inhibition of EPSCs, more P/Q-type Ca(2+) channels were involved in GABA(B) receptor-mediated inhibition of IPSCs. Finally, baclofen blocked the frequency-dependent depression of EPSCs and IPSCs, but was less effective at blocking frequency-dependent facilitation of EPSCs. Our results demonstrate that presynaptic GABA(B) receptors are expressed on the terminals of both excitatory and inhibitory synapses onto CA3 interneurons and that their activation modulates essential components of the release process underlying transmission at these two synapse types. PMID- 12527731 TI - Activation of mGlu receptors induces LTD without affecting postsynaptic sensitivity of CA1 neurons in rat hippocampal slices. AB - Two forms of long-term depression (LTD) of excitatory synaptic transmission have been identified in the mammalian CNS, which are induced by the synaptic activation of N-methyl-D-aspartate (NMDA) and metabotropic glutamate (mGlu) receptors, respectively. The mGlu receptor-dependent form of LTD can be activated by application of 3,5-dihydroxyphenylglycine (DHPG), a group I selective mGlu receptor agonist. DHPG-induced LTD is increasingly being used to investigate the mechanisms of mGlu receptor-dependent LTD. However, recent experiments have argued for both a pre- and postsynaptic locus of expression of DHPG-induced LTD. In the present study we report that DHPG-induced LTD is not associated with changes in the sensitivity of CA1 neurons to bath applied AMPA. Furthermore, in contrast to homosynaptic LTD, DHPG-induced LTD is also not associated with changes in sensitivity to focally uncaged L-glutamate. These data do not support the notion that DHPG-induced LTD requires a modification of AMPA receptors, such as their internalisation, but are compatible with a presynaptic mechanism of expression. PMID- 12527732 TI - Angiotensin II AT1 receptor stimulates Na+ -K+ATPase activity through a pathway involving PKC-zeta in rat thyroid cells. AB - Angiotensin II (Ang II) receptor subtype 1, AT1, is expressed by the rat thyroid. A relationship between thyroid function and several components of the renin angiotensin system has also been established, but the Ang II cellular effects in thyrocytes and its transduction signalling remain undefined. The aim of the present paper was to investigate the modulation of the activity of the Na(+) K(+)ATPase by Ang II and its intracellular transduction pathway in PC-Cl3 cells, an established epithelial cell line derived from rat thyroid. Here we have demonstrated, by RT-PCR analysis, the expression of mRNA for the Ang II AT1 receptor in PC-Cl3 cells; mRNA for the Ang II AT2 receptor was not detected. Ang II was not able to affect the intracellular Ca(2+) concentration in fura-2-loaded cells, but it stimulated the translocation from the cytosol to the plasma membrane of atypical protein kinase C-zeta (PKC-zeta) and -iota (PKC-) isoforms with subsequent phosphorylation of the extracellular signal-regulated kinases 1 and 2 (ERK1 and 2). Translocated atypical PKCs displayed temporally different activations, the activation of PKC-zeta being the fastest. PC-Cl3 cells stimulated with increasing Ang II concentrations showed dose- and time-dependent activation of the Na(+)-K(+)ATPase activity, which paralleled the PKC-zeta translocation time course. Na(+)-K(+)ATPase activity modulation was dependent on PKC activation since the PKC antagonist staurosporine abolished the stimulatory effect of Ang II. The inhibition of the ERK kinases 1 and 2 (MEK1 and 2) by PD098059 (2'-amino-3'-methoxyflavone) failed to block the effect of Ang II on the Na(+)-K(+)ATPase activity. In conclusion, our results suggest that Ang II modulates Na(+)-K(+)ATPase activity in PC-Cl3 cells through the AT1 receptor via activation of atypical PKC-zeta while the Ang II-activated PKC- appears to have other as yet unknown functions. PMID- 12527733 TI - Effects of osmotic changes on the chemoreceptor cell of rat carotid body. AB - The carotid body plays a crucial role in cardiorespiratory regulation. In the present study we investigated the effect of osmotic changes on cytoplasmic calcium concentration ([Ca(2+)](c)) and pH (pH(i)) of isolated chemoreceptor cells of the rat carotid body. In CO(2)/HCO(3)(-)-buffered medium, reduction of osmolality from the control level of 300 mosmol kg(-1) to 250-285 mosmol kg(-1) resulted in a rise in [Ca(2+)](c), as measured with Indo-1, whereas elevation of osmolality to 350 mosmol kg(-1) had no effect. The Ca(2+) response required extracellular Ca(2+) and was reduced by application of the L-type Ca(2+) channel antagonist nifedipine (10 microM). The hyposmosis-induced Ca(2+) response could be prevented by application of niflumic acid (300 microM), an inhibitor of the swelling-activated Cl(-) channel. In whole-cell patch-clamp experiments niflumic acid abolished the swelling-activated Cl(-) current but only slightly depressed the Ca(2+) current. The inhibition of Ca(2+) current by niflumic acid does not account for its action in preventing of hyposmosis-induced Ca(2+) response, which seems to be initiated by Cl(-)-mediated depolarisation. Withdrawal of CO(2)/HCO(3)(-) also prevented the Ca(2+) response. Reduction of the osmotic concentration by 50 mosmol kg(-1) induced a small but sustained decrease in pH(i), while elevation by 50 mosmol kg(-1) had an inverse effect, as measured fluorimetrically with carboxy SNARF-1. Our conclusion is that in the rat chemoreceptor cell the activation of Cl(-) channels, e.g. by hyposmotic challenge, induces depolarisation, which, in turn, activates voltage-gated Ca(2+) channels. PMID- 12527734 TI - Parallel decrease in omega-conotoxin-sensitive transmission and dopamine-induced inhibition at the striatal synapse of developing rats. AB - Whole-cell patch-clamp recordings of GABAergic IPSCs were made from cholinergic interneurones in slices of striatum from developing rats aged 21-60 days postnatal. In addition, the Ca(2+) channel subtypes involved in synaptic transmission, as well as dopamine (DA)-induced presynaptic inhibition, were investigated pharmacologically with development by bath application of Ca(2+) channel blockers and DA receptor agonists. The IPSC amplitude was reduced by omega-conotoxin GVIA (omega-CgTX) or omega-agatoxin TK (omega-Aga-TK) across the whole age range, suggesting that multiple types of Ca(2+) channels mediate transmission of the synapse. The IPSC fraction reduced by omega-CgTX significantly decreased, whereas that reduced by omega-Aga-TK remained unchanged with development. DA or quinpirole, a D(2)-like receptor agonist, presynaptically reduced the IPSC amplitude throughout development. The DA-induced inhibition decreased with age in parallel with the decrease in N-type Ca(2+) channels. DA showed no further inhibition of IPSCs after the inhibitory effect of omega-CgTX had reached steady state throughout development. These results demonstrate that there is a functional link between presynaptic N-type Ca(2+) channels and D(2) like DA receptors at inhibitory synapses in the striatum. They also demonstrate that the suppression of GABAergic transmission by D(2)-like receptors is mediated by modulation of N-type Ca(2+) channels and decreases in parallel with the developmental decline in the contribution of N-type Ca(2+) channels to exocytosis. PMID- 12527735 TI - Acute exposure to thyroid hormone increases Na+ current and intracellular Ca2+ in cat atrial myocytes. AB - Whole-cell recording methods and fluorescence microscopy were used to study the effects of acute exposure to thyroid hormone (T(3)) on cat atrial myocytes. Acute exposure ( approximately 5 min) to 10 nM T(3) significantly increased tetrodotoxin (TTX)-sensitive inward Na(+) current (I(Na)) at voltages between -40 and +20 mV. At maximal I(Na) activation (-40 mV) T(3) increased peak I(Na) by 32 %. T(3) had no effect on the time course of I(Na) decay, voltage dependence of activation, inactivation, or recovery from inactivation. Comparable exposures to reverse T(3) (rT(3)) or T(4) had no effect on I(Na). L-type Ca2+ current was unaffected by acute exposure to T(3). T(3)-induced increases in I(Na) were unaffected by 50 microM nickel, a blocker of T-type Ca2+ current. T(3) significantly increased cell shortening (+62 %) and could elicit spontaneous action potentials arising from Ca2+ -mediated after-depolarizations. T(3) (but not rT(3)) significantly increased baseline intracellular Ca2+, release of Ca2+ from sarcoplasmic reticulum (SR) and caffeine (10 mM)-induced release of SR Ca2+. We conclude that acute T(3) exposure increases Na(+) influx via I(Na) and thereby stimulates reverse-mode Na(+)-Ca2+ exchange to increase intracellular Ca2+ content and release. As a result, T(3) increases contraction strength, and can initiate Ca2+ -mediated arrhythmic activity. Acute non-genomic effects of T(3) can contribute to the positive inotropy and sinus (atrial) tachycardia traditionally attributed to chronic, genomic effects of elevated thyroid hormone on atrial muscle. PMID- 12527736 TI - Streptomycin and intracellular calcium modulate the response of single guinea-pig ventricular myocytes to axial stretch. AB - We tested the hypothesis that both stretch-activated channels (SACs) and intracellular calcium ([Ca(2+)](i)) are important in the electrical response of single guinea-pig ventricular myocytes to axial stretch. Myocytes were attached to carbon fibre transducers and stretched, sarcomere length increased by approximately 9 %, and there was a prolongation of the action potential duration. Streptomycin, a blocker of SACs, had no effect upon the shortening, [Ca(2+)](i) transients or action potentials of electrically stimulated, unstretched myocytes, at a concentration of 50 microM, but at 40 microM, prevented any stretch-induced increase in action potential duration. Under action potential clamp, stretch elicited a current with a linear current-voltage relationship that was inward at membrane potentials negative to its reversal potential of -30 mV, in 10 of 24 cells tested, and was consistent with the activation of non-specific, cationic SACs. This current was not seen in any stretched cells that were exposed to 40 microM streptomycin. However, exposure of cells to 5 microM BAPTA-AM, in order to reduce [Ca(2+)](i) transients, also abolished stretch-induced prolongation of the action potential. We conclude that both SACs and [Ca(2+)](i) are important in the electrical response of cardiac myocytes to stretch, and propose that stretch induced changes in electrical activity and [Ca(2+)](i) may be linked by inter dependent mechanisms. PMID- 12527737 TI - Chick hair cells do not exhibit voltage-dependent somatic motility. AB - It is generally believed that mechanical amplification by cochlear hair cells is necessary to enhance the sensitivity and frequency selectivity of hearing. In the mammalian ear, the basis of cochlear amplification is believed to be the voltage dependent electromotility of outer hair cells (OHCs). The avian basilar papilla contains tall and short hair cells, with the former being comparable to inner hair cells, and the latter comparable to OHCs, based on their innervation patterns. In this study, we sought evidence for somatic electromotility by direct measurements of voltage-dependent length changes in both tall and short hair cells at nanometre resolution. Microchamber and whole-cell voltage-clamp techniques were used. Motility was measured with a photodiode-based measurement system. Non-linear capacitance, an electrical signature of somatic motility, was also measured to complement motility measurement. Significantly, chick hair cells did not exhibit somatic motility nor express non-linear capacitance. The lack of somatic motility suggests that in avian hair cells the active process resides elsewhere, most likely in the hair cell stereocilia. PMID- 12527738 TI - Does nitric oxide allow endothelial cells to sense hypoxia and mediate hypoxic vasodilatation? In vivo and in vitro studies. AB - Hypoxia-evoked vasodilatation is a fundamental regulatory mechanism that is often attributed to adenosine. The identity of the O(2) sensor is unknown. Nitric oxide (NO) inhibits endothelial mitochondrial respiration and ATP generation by competing with O(2) for its binding site on cytochrome oxidase. We proposed that in vivo this interaction allows endothelial cells to release adenosine when O(2) tension falls or NO concentration increases. Using anaesthetised rats, we confirmed that the increase in femoral vascular conductance (FVC, hindlimb vasodilatation) evoked by systemic hypoxia is attenuated by NO synthesis blockade with L-NAME, but restored when baseline FVC is restored by infusion of NO donor. This "restored" hypoxic response, like the control hypoxic response, is inhibited by the adenosine A(1) receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine. Similarly, the FVC increase evoked by adenosine infusion was attenuated by L-NAME but restored by infusion of NO donor. However, when baseline FVC was restored after L-NAME with 8-bromo-cGMP, the FVC increase evoked by adenosine infusion was restored, but not in response to systemic hypoxia, suggesting that adenosine was no longer released by hypoxia. Infusion of NO donor at a given rate after treatment with L-NAME evoked a greater FVC increase during systemic hypoxia than during normoxia, both responses being reduced by 8-cyclopentyl-1,3 dipropylxanthine. Finally, both bradykinin and NO donor released adenosine from superfused endothelial cells in vitro; L-NAME attenuated only the former response. We propose that in vivo, shear-released NO increases the apparent K(m) of endothelial cytochrome oxidase for O(2), allowing the endothelium to act as an O(2) sensor, releasing adenosine in response to moderate falls in O(2). PMID- 12527739 TI - Muscle fractal vascular branching pattern and microvascular perfusion heterogeneity in endurance-trained and untrained men. AB - Less heterogeneous skeletal muscle perfusion has recently been reported in endurance-trained compared to untrained men at macrovascular level. The causes of this difference in perfusion heterogeneity are unknown as is whether the same difference is observed in microvasculature. We hypothesised that the difference could be caused by changes in muscle vascular branching pattern. Perfusion was measured in resting and exercising muscle in 14 endurance-trained and seven untrained men using [(15)O]water and positron emission tomography. Fractal dimension (D) of perfusion distribution was calculated as a measure of fractal characteristics of muscle vascular branching pattern. Perfusion heterogeneity in microvascular units (1 mm(3) samples) was estimated using the measured heterogeneity in voxels of positron emission tomography (PET) images (relative dispersion, RD = S.D./mean) and corresponding D values. D was similar between the groups (exercising muscle 1.11 +/- 0.07 and 1.14 +/- 0.06, resting muscle 1.12 +/ 0.06 and 1.14 +/- 0.03, trained and untrained, respectively). Trained men had lower perfusion (151 +/- 44 vs. 218 +/- 87 ml min(-1) kg(-1), P < 0.05) and macrovascular perfusion heterogeneity (relative dispersion 21 +/- 5 vs. 25 +/- 5 %, P < 0.05) in exercising muscle than untrained men. Furthermore, estimated perfusion heterogeneity in microvascular units in exercising muscle was also lower in trained men (33 +/- 7 vs.48 +/- 19 %, P < 0.05). These results show that fractal vascular branching pattern is similar in endurance-trained and untrained men but perfusion is less heterogeneous at both the macro- and the microvascular level in endurance-trained men. Thus, changes in fractal branching pattern do not explain the differences in perfusion heterogeneity between endurance-trained and untrained men. PMID- 12527740 TI - Reduced contraction strength with increased intracellular [Ca2+] in left ventricular trabeculae from failing rat hearts. AB - Intracellular calcium ([Ca2+](i)) and isometric force were measured in left ventricular (LV) trabeculae from spontaneously hypertensive rats (SHR) with failing hearts and normotensive Wistar-Kyoto (WKY) controls. At a physiological stimulation frequency (5 Hz), and at 37 degrees C, the peak stress of SHR trabeculae was significantly (P < or = 0.05) reduced compared to WKY (8 +/- 1 mN mm(-2) (n = 8) vs. 21 +/- 5 mN mm(-2) (n = 8), respectively). No differences between strains in either the time-to-peak stress, or the time from peak to 50 % relaxation were detected. Measurements using fura-2 showed that in the SHR both the peak of the Ca2+ transient and the resting [Ca2+](i) were increased compared to WKY (peak: 0.69 +/- 0.08 vs. 0.51 +/- 0.08 microM(P < or = 0.1) and resting: 0.19 +/- 0.02 vs. 0.09 +/- 0.02 microM(P < or = 0.05), SHR vs. WKY, respectively). The decay of the Ca2+ transient was prolonged in SHR, with time constants of: 0.063 +/- 0.002 vs. 0.052 +/- 0.003 s (SHR vs. WKY, respectively). Similar results were obtained at 1 Hz stimulation, and for [Ca2+ ](o) between 0.5 and 5 mM. The decay of the caffeine-evoked Ca2+ transient was slower in SHR (9.8 +/- 0.7 s (n = 8) vs. 7.7 +/- 0.2 s (n = 8) in WKY), but this difference was removed by use of the SL Ca2+ -ATPase inhibitor carboxyeosin. Histological examination of transverse sections showed that the fractional content of perimysial collagen was increased in SHR compared to WKY (18.0 +/- 4.6 % (n = 10) vs. 2.9 +/- 0.9 % (n = 11) SHR vs. WKY, respectively). Our results show that differences in the amplitude and the time course of the Ca2+ transient between SHR and WKY do not explain the reduced contractile performance of SHR myocardium per se. Rather, we suggest that, in this animal model of heart failure, contractile function is compromised by increased collagen, and its three dimensional organisation, and not by reduced availability of intracellular Ca2+. PMID- 12527741 TI - Uptake of locally applied deoxyglucose, glucose and lactate by axons and Schwann cells of rat vagus nerve. AB - We asked whether, in a steady state, neurons and glial cells both take up glucose sufficient for their energy requirements, or whether glial cells take up a disproportionate amount and transfer metabolic substrate to neurons. A desheathed rat vagus nerve was held crossways in a laminar flow perfusion chamber and stimulated at 2 Hz. (14)C-labelled substrate was applied from a micropipette for 5 min over a < 0.6 mm band of the surface of the nerve. After 10-55 min incubation, the nerve was lyophilized and the longitudinal distribution of radioactivity measured. When the weakly metabolizable analogue of glucose, 2 deoxy-[U-(14)C]D-glucose (*DG), was applied, the profiles of the radioactivity broadened with time, reaching distances several times the mean length of the Schwann cells (0.32 mm; most of the Schwann cells are non-myelinating). The profiles were well fitted by curves calculated for diffusion in a single compartment, the mean diffusion coefficient being 463 +/- 34 microm(2) s(-1) (+/- S.E.M., n = 16). Applications of *DG were repeated in the presence of the gap junction blocker, carbenoxolone (100 microM). The profiles were now narrower and better fitted with two compartments. One compartment had a coefficient not significantly different from that in the absence of the gap junction blocker (axons), the other compartment had a coefficient of 204 +/- 24 microm(2) s(-1), n = 4. Addition of the gap junction blocker 18-alpha-glycyrrhetinic acid, or blocking electrical activity with TTX, also reduced longitudinal diffusion. Ascribing the compartment in which diffusion was reduced by these treatments to non-myelinating Schwann cells, we conclude that 78.0 +/- 3.6 % (n = 9) of the uptake of *DG was into Schwann cells. This suggests that there was transfer of metabolic substrate from Schwann cells to axons. Local application of [(14)C]glucose or [(14)C]lactate led to variable labelling along the length of the nerve, but with both substrates narrow peaks were often present at the application site; these were greatly reduced by subsequent treatment with amylase, a glycogen-degrading enzyme. PMID- 12527742 TI - Sensory and electrophysiological properties of guinea-pig sensory neurones expressing Nav 1.7 (PN1) Na+ channel alpha subunit protein. AB - The TTX-sensitive Na(v)1.7 (PN1) Na(+) channel alpha subunit protein is expressed mainly in small dorsal root ganglion (DRG) neurones. This study examines immunocytochemically whether it is expressed exclusively or preferentially in nociceptive primary afferent DRG neurones, and determines the electrophysiological properties of neurones that express it. Intracellular somatic action potentials (APs) evoked by dorsal root stimulation were recorded in L6/S1 DRG neurones at 30 +/- 2 degrees C in vivo in deeply anaesthetised young guinea-pigs. Each neurone was classified, from its dorsal root conduction velocity (CV) as a C-, Adelta- or Aalpha/beta-fibre unit and from its response to mechanical and thermal stimuli, as a nociceptive, low threshold mechanoreceptive (LTM) or unresponsive unit. Fluorescent dye was injected into the soma and Na(v)1.7-like immunoreactivity (Na(v)1.7-LI) was examined on sections of dye injected neurones. All C-, 90 % of Adelta- and 40 % of Aalpha/beta-fibre units, including both nociceptive and LTM units, showed Na(v)1.7-LI. Positive units included 1/1 C-LTM, 6/6 C-nociceptive, 4/4 C-unresponsive (possible silent nociceptive) units, 5/6 Adelta-LTM (D hair), 13/14 Adelta-nociceptive, 2/9 Aalpha/beta-nociceptive, 10/18 Aalpha/beta-LTM cutaneous and 0/9 Aalpha/beta muscle spindle afferent units. Overall, a higher proportion of nociceptive than of LTM neurones was positive, and the median relative staining intensity was greater in nociceptive than LTM units. Na(v)1.7-LI intensity was clearly positively correlated with AP duration and (less strongly) negatively correlated with CV and soma size. Since nociceptive units tend overall to have longer duration APs, slower CVs and smaller somata, these correlations may be related to the generally greater expression of Na(v)1.7 in nociceptive units. PMID- 12527743 TI - NOS inhibition increases bubble formation and reduces survival in sedentary but not exercised rats. AB - Previously we have shown that chronic as well as a single bout of exercise 20 h prior to a simulated dive protects rats from severe decompression illness (DCI) and death. However, the mechanism behind this protection is still not known. The present study determines the effect of inhibiting nitric oxide synthase (NOS) on bubble formation in acutely exercised and sedentary rats exposed to hyperbaric pressure. A total of 45 adult female Sprague-Dawley rats (270-320 g) were randomly assigned into exercise or sedentary control groups, with and without NOS inhibition, using L-NAME (0.05 or 1 mg ml(-1)) (a nonselective NOS inhibitor). Exercising rats ran intervals on a treadmill for 1.5 h, 20 h prior to the simulated dive. Intervals alternated between 8 min at 85-90 % of maximal oxygen uptake, and 2 min at 50-60 %. Rats were compressed (simulated dive) in a pressure chamber, at a rate of 200 kPa min(-1) to a pressure of 700 kPa, and maintained for 45 min breathing air. At the end of the exposure period, rats were decompressed linearly to the "surface" (100 kPa) at a rate of 50 kPa min(-1). Immediately after reaching the surface the animals were anaesthetised and the right ventricle was insonated using ultrasound. The study demonstrated that sedentary rats weighing more than 300 g produced a large amount of bubbles, while those weighing less than 300 g produced few bubbles and most survived the protocol. Prior exercise reduced bubble formation and increased survival in rats weighing more than 300 g, confirming the results from the previous study. During NOS inhibition, the simulated dive induced significantly more bubbles in all sedentary rats weighing less than 300 g. However, this effect could be attenuated by a single bout of exercise 20 h before exposure. The present study demonstrates two previously unreported findings: that administration of L-NAME allows substantial bubble formation and decreased survival in sedentary rats, and that a single bout of exercise protects NOS-inhibited rats from severe bubble formation and death. This is the first report to indicate that biochemical processes are involved in bubble formation, and this information may be important in the search for preventive measures for and treatment of DCI. PMID- 12527744 TI - Beta(2)-adrenergic receptor polymorphism and nitric oxide-dependent forearm blood flow responses to isoproterenol in humans. AB - Polymorphisms in the gene encoding the beta(2)-adrenoceptor have been associated with interindividual differences in blood pressure and the diagnosis of hypertension. A common polymorphism resulting in a change from arginine to glycine at amino acid 16 (Arg16-->Gly) enhances agonist-promoted downregulation of receptor expression in vitro. It is unknown whether genotype-dependent differences in nitric oxide generation contribute to differences in vasodilator responses to beta(2)-agonists in vivo. To address this question, venous occlusion plethysmography was used to measure forearm blood flow responses to graded brachial artery infusions of the beta-agonist isoproterenol in 41 healthy normotensive Caucasian adults (mean age (+/- S.D.) = 29 +/- 6 years), who were either Arg16 (n = 18) or Gly16 (n = 23) homozygotes. Compared to Arg16 homozygotes, Gly16 homozygotes demonstrated significantly greater blood flow responses to isoproterenol (P = 0.02). After inhibition of nitric oxide synthase by N(gamma)-monomethyl-L-arginine, blood flow responses did not differ significantly between genotype groups (P = 0.27). Consequently, effects of the Arg16 Gly polymorphism on forearm blood flow responses to isoproterenol appear to be dependent on differences in endothelial generation of nitric oxide. In contrast to previous reports based on systemic infusions of beta(2)-agonists, our findings indicate that regional blood flow responses to locally infused isoproterenol are significantly greater in Gly16 than in Arg16 homozygotes. PMID- 12527745 TI - Mechanisms of acute natriuresis in normal humans on low sodium diet. AB - This study evaluates the relative importance of several mechanisms possibly involved in the natriuresis elicited by slow sodium loading, i.e. the renin angiotensin-aldosterone system (RAAS), mean arterial blood pressure (MAP), glomerular filtration rate (GFR), atrial natriuretic peptide (ANP), oxytocin and nitric oxide (NO). Eight seated subjects on standardised sodium intake (30 mmol NaCl day(-1)) received isotonic saline intravenously (NaLoading: 20 micromol Na(+) kg(-1) min(-1) or approximately 11 ml min(-1) for 240 min). NaLoading did not change MAP or GFR (by clearance of (51)Cr-EDTA). Significant natriuresis occurred within 1 h (from 9 +/- 3 to 13 +/- 2 micromol min(-1)). A 6-fold increase was found during the last hour of infusion as plasma renin activity, angiotensin II (ANGII) and aldosterone decreased markedly. Sodium excretion continued to increase after NaLoading. During NaLoading, plasma renin activity and ANGII were linearly related (R = 0.997) as were ANGII and aldosterone (R = 0.999). The slopes were 0.40 pM ANGII (mi.u. renin activity)(-1) and 22 pM aldosterone (pM ANGII)(-1). Plasma ANP and oxytocin remained unchanged, as did the urinary excretion rates of cGMP and NO metabolites (NO(x)). In conclusion, sodium excretion may increase 7-fold without changes in MAP, GFR, plasma ANP, plasma oxytocin, and cGMP- and NO(x) excretion, but concomitant with marked decreases in circulating RAAS components. The immediate renal response to sodium excess appears to be fading of ANGII-mediated tubular sodium reabsorption. Subsequently the decrease in aldosterone may become important. PMID- 12527746 TI - Age and sex differences in human motor cortex input-output characteristics. AB - Stimulus-response curves for motor evoked potentials (MEPs) induced in a hand muscle by transcranial magnetic stimulation (TMS) were constructed for 42 subjects with the aim of identifying differences related to age and sex. There was no effect of age on the resting threshold to TMS, the maximal amplitude of the MEP that could be evoked (MEP(max)) or the maximal slope of the stimulus response curve. However, higher stimulus intensities were required to achieve both MEP(max) and the maximal slope in the older subjects. The trial-to-trial variability of MEPs was greater in the older subjects, particularly at intensities near threshold. There was a significant interaction between age, threshold and trial-to-trial variability of MEP amplitude. Overall, MEP variability fell markedly as stimulus intensity increased above threshold but less rapidly in older than in younger subjects. Females tended to have larger MEP variability than males, but age and threshold were much stronger modulators than sex. These differences in input-output characteristics are likely to be due either to a decreased number of spinal motoneurones being activated synchronously in older subjects, or to the activation of the same number of motoneurones in a less synchronous manner, leading to phase cancellation in the surface electromyogram. PMID- 12527747 TI - Vestibular actions on back and lower limb muscles during postural tasks in man. AB - The vestibular system was activated by galvanic electrical stimulation in 19 normal subjects. With the head turned to one side so that the stimulating anode was on the posterior mastoid process, stimulation caused standing subjects to sway backwards in the sagittal plane. Electromyography showed bilateral activation of erector spinae, gluteus maximus, biceps femoris, soleus and intrinsic foot (toe flexor) muscles. When head direction or electrode polarity was reversed so that the anode was anterior, all those muscles became less active and the subjects swayed forwards. With the head facing forward, stimulation caused sideways sway in the coronal plane, towards the anode, with excitation of the erector spinae on the anode side and reduced activity on the cathode side. The limb muscles were activated on the side opposite the anode and showed complex responses on the anode side. Responses were detectable in the erectores spinae muscles in sitting subjects. No responses in limb muscles were detected in the sitting posture. Subject responses in erector spinae recorded at L3/L4 had latencies from 59 to 110 ms, using a 2 mA stimulus. Latencies in lower limb muscles were longer. The results suggest a role for the vestibular system and descending brain stem motor pathways to the erectores spinae muscles in the control of postural orientation of the back when sitting and standing. The conduction velocity in the motor pathway was estimated to be 13 +/- 10 m s(-1) (mean +/- S.D., n = 12 subjects). PMID- 12527748 TI - Tenascin-C signaling through induction of 14-3-3 tau. AB - We searched by a cDNA subtraction screen for differentially expressed transcripts in MCF-7 mammary carcinoma cells grown on tenascin-C versus fibronectin. On tenascin-C, cells had irregular shapes with many processes, whereas on fibronectin they were flat with a cobble stone-like appearance. We found elevated levels of 14-3-3 tau transcripts and protein in cells grown on tenascin-C. To investigate the consequences of an increased level of this phospho serine/threonine-binding adaptor protein, we transfected MCF-7 cells with a construct encoding full-length 14-3-3 tau protein and selected clones with the highest expression levels. The morphology of these cells on tenascin-C was flat, resembling that of cells on fibronectin. This was reflected by a similar pattern of F-actin staining on either substratum. Furthermore, the growth rate on tenascin-C was increased compared with the parental cells. After transient transfection of HT1080 fibrosarcoma and T98G glioblastoma cells with 14-3-3 tau, only the 14-3-3 tau-expressing cells were able to adhere and survive on tenascin C, whereas all cells adhered well on fibronectin. Therefore, we postulate that tenascin-C promotes the growth of tumor cells by causing an increase in the expression of 14-3-3 tau, which in turn has a positive effect on tumor cell adhesion and growth. PMID- 12527749 TI - Spatio-temporal activation of caspase revealed by indicator that is insensitive to environmental effects. AB - Indicator molecules for caspase-3 activation have been reported that use fluorescence resonance energy transfer (FRET) between an enhanced cyan fluorescent protein (the donor) and enhanced yellow fluorescent protein (EYFP; the acceptor). Because EYFP is highly sensitive to proton (H+) and chloride ion (Cl-) levels, which can change during apoptosis, this indicator's ability to trace the precise dynamics of caspase activation is limited, especially in vivo. Here, we generated an H+- and Cl--insensitive indicator for caspase activation, SCAT, in which EYFP was replaced with Venus, and monitored the spatio-temporal activation of caspases in living cells. Caspase-3 activation was initiated first in the cytosol and then in the nucleus, and rapidly reached maximum activation in 10 min or less. Furthermore, the nuclear activation of caspase-3 preceded the nuclear apoptotic morphological changes. In contrast, the completion of caspase-9 activation took much longer and its activation was attenuated in the nucleus. However, the time between the initiation of caspase-9 activation and the morphological changes was quite similar to that seen for caspase-3, indicating the activation of both caspases occurred essentially simultaneously during the initiation of apoptosis. PMID- 12527750 TI - Binding of an ankyrin-1 isoform to obscurin suggests a molecular link between the sarcoplasmic reticulum and myofibrils in striated muscles. AB - Assembly of specialized membrane domains, both of the plasma membrane and of the ER, is necessary for the physiological activity of striated muscle cells. The mechanisms that mediate the structural organization of the sarcoplasmic reticulum with respect to the myofibrils are, however, not known. We report here that ank1.5, a small splice variant of the ank1 gene localized on the sarcoplasmic reticulum membrane, is capable of interacting with a sequence of 25 aa located at the COOH terminus of obscurin. Obscurin is a giant sarcomeric protein of approximately 800 kD that binds to titin and has been proposed to mediate interactions between myofibrils and other cellular structures. The binding sites and the critical aa required in the interaction between ank1.5 and obscurin were characterized using the yeast two-hybrid system, in in vitro pull-down assays and in experiments in heterologous cells. In differentiated skeletal muscle cells, a transfected myc-tagged ank1.5 was found to be selectively restricted near the M line region where it colocalized with endogenous obscurin. The M line localization of ank1.5 required a functional obscurin-binding site, because mutations of this domain resulted in a diffused distribution of the mutant ank1.5 protein in skeletal muscle cells. The interaction between ank1.5 and obscurin represents the first direct evidence of two proteins that may provide a direct link between the sarcoplasmic reticulum and myofibrils. In keeping with the proposed role of obscurin in mediating an interaction with ankyrins and sarcoplasmic reticulum, we have also found that a sequence with homology to the obscurin-binding site of ank1.5 is present in the ank2.2 isoform, which in striated muscles has been also shown to associate with the sarcoplasmic reticulum. Accordingly, a peptide containing the COOH terminus of ank2.2 fused with GST was found to bind to obscurin. Based on reported evidence showing that the COOH terminus of ank2.2 is necessary for the localization of ryanodine receptors and InsP3 receptors in the sarcoplasmic reticulum, we propose that obscurin, through multiple interactions with ank1.5 and ank2.2 isoforms, may assemble a large protein complex that, in addition to a structural function, may play a role in the organization of specific subdomains in the sarcoplasmic reticulum. PMID- 12527751 TI - Compensation mechanism in tumor cell migration: mesenchymal-amoeboid transition after blocking of pericellular proteolysis. AB - Invasive tumor dissemination in vitro and in vivo involves the proteolytic degradation of ECM barriers. This process, however, is only incompletely attenuated by protease inhibitor-based treatment, suggesting the existence of migratory compensation strategies. In three-dimensional collagen matrices, spindle-shaped proteolytically potent HT-1080 fibrosarcoma and MDA-MB-231 carcinoma cells exhibited a constitutive mesenchymal-type movement including the coclustering of beta 1 integrins and MT1-matrix metalloproteinase (MMP) at fiber bindings sites and the generation of tube-like proteolytic degradation tracks. Near-total inhibition of MMPs, serine proteases, cathepsins, and other proteases, however, induced a conversion toward spherical morphology at near undiminished migration rates. Sustained protease-independent migration resulted from a flexible amoeba-like shape change, i.e., propulsive squeezing through preexisting matrix gaps and formation of constriction rings in the absence of matrix degradation, concomitant loss of clustered beta 1 integrins and MT1-MMP from fiber binding sites, and a diffuse cortical distribution of the actin cytoskeleton. Acquisition of protease-independent amoeboid dissemination was confirmed for HT-1080 cells injected into the mouse dermis monitored by intravital multiphoton microscopy. In conclusion, the transition from proteolytic mesenchymal toward nonproteolytic amoeboid movement highlights a supramolecular plasticity mechanism in cell migration and further represents a putative escape mechanism in tumor cell dissemination after abrogation of pericellular proteolysis. PMID- 12527752 TI - Direct visualization of Ras proteins in spatially distinct cell surface microdomains. AB - Localization of signaling complexes to specific microdomains coordinates signal transduction at the plasma membrane. Using immunogold electron microscopy of plasma membrane sheets coupled with spatial point pattern analysis, we have visualized morphologically featureless microdomains, including lipid rafts, in situ and at high resolution. We find that an inner-plasma membrane lipid raft marker displays cholesterol-dependent clustering in microdomains with a mean diameter of 44 nm that occupy 35% of the cell surface. Cross-linking an outer leaflet raft protein results in the redistribution of inner leaflet rafts, but they retain their modular structure. Analysis of Ras microlocalization shows that inactive H-ras is distributed between lipid rafts and a cholesterol-independent microdomain. Conversely, activated H-ras and K-ras reside predominantly in nonoverlapping, cholesterol-independent microdomains. Galectin-1 stabilizes the association of activated H-ras with these nonraft microdomains, whereas K-ras clustering is supported by farnesylation, but not geranylgeranylation. These results illustrate that the inner plasma membrane comprises a complex mosaic of discrete microdomains. Differential spatial localization within this framework can likely account for the distinct signal outputs from the highly homologous Ras proteins. PMID- 12527753 TI - Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development. AB - Mitochondrial morphology is determined by a dynamic equilibrium between organelle fusion and fission, but the significance of these processes in vertebrates is unknown. The mitofusins, Mfn1 and Mfn2, have been shown to affect mitochondrial morphology when overexpressed. We find that mice deficient in either Mfn1 or Mfn2 die in midgestation. However, whereas Mfn2 mutant embryos have a specific and severe disruption of the placental trophoblast giant cell layer, Mfn1-deficient giant cells are normal. Embryonic fibroblasts lacking Mfn1 or Mfn2 display distinct types of fragmented mitochondria, a phenotype we determine to be due to a severe reduction in mitochondrial fusion. Moreover, we find that Mfn1 and Mfn2 form homotypic and heterotypic complexes and show, by rescue of mutant cells, that the homotypic complexes are functional for fusion. We conclude that Mfn1 and Mfn2 have both redundant and distinct functions and act in three separate molecular complexes to promote mitochondrial fusion. Strikingly, a subset of mitochondria in mutant cells lose membrane potential. Therefore, mitochondrial fusion is essential for embryonic development, and by enabling cooperation between mitochondria, has protective effects on the mitochondrial population. PMID- 12527754 TI - Domain motions and quaternary packing of phosphofructokinase-2 from Escherichia coli studied by small angle x-ray scattering and homology modeling. AB - The binding of MgATP and fructose-6-phosphate to phosphofructokinase-2 from Escherichia coli induces conformational changes that result in significant differences in the x-ray-scattering profiles compared with the unligated form of the enzyme. When fructose- 6-phosphate binds to the active site of the enzyme, the pair distribution function exhibits lower values at higher distances, indicating a more compact structure. Upon binding of MgATP to the allosteric site of the enzyme, the intensity at lower angles increases as a consequence of tetramer formation, but differences along higher angles also suggest changes at the tertiary structure level. We have used homology modeling to build the native dimeric form of phosphofructokinase-2 and fitted the experimental scattering curves by using rigid body movements of the domains in the model, similar to those observed in known homologous structures. The best fit with the experimental data of the unbound protein was achieved with open conformations of the domains in the model, whereas domain closure improves the agreement with the scattering of the enzyme-fructose-6-phosphate complex. Using the same approach, we utilized the scattering curve of the phosphofructokinase-2-MgATP complex to model the arrangement and conformation of dimers in the tetramer. We observed that, along with tetramerization, binding of MgATP to the allosteric site induces domain closure. Additionally, we used the scattering data to restore the low resolution structure of phosphofructokinase-2 (free and bound forms) by an ab initio procedure. Based on these findings, a proposal is made to account for the inhibitory effect of MgATP on the enzymatic activity. PMID- 12527755 TI - Nod2 is a general sensor of peptidoglycan through muramyl dipeptide (MDP) detection. AB - Nod2 activates the NF-kappaB pathway following intracellular stimulation by bacterial products. Recently, mutations in Nod2 have been shown to be associated with Crohn's disease, suggesting a role for bacteria-host interactions in the etiology of this disorder. We show here that Nod2 is a general sensor of peptidoglycan through the recognition of muramyl dipeptide (MDP), the minimal bioactive peptidoglycan motif common to all bacteria. Moreover, the 3020insC frameshift mutation, the most frequent Nod2 variant associated with Crohn's disease patients, fully abrogates Nod2-dependent detection of peptidoglycan and MDP. Together, these results impact on the understanding of Crohn's disease development. Additionally, the characterization of Nod2 as the first pathogen recognition molecule that detects MDP will help to unravel the well known biological activities of this immunomodulatory compound. PMID- 12527756 TI - MTA1 interacts with MAT1, a cyclin-dependent kinase-activating kinase complex ring finger factor, and regulates estrogen receptor transactivation functions. AB - The transcriptional activity of estrogen receptor-alpha is controlled by coregulators. MTA1 (metastasis-associated protein 1) represses estrogen receptor alpha-driven transcription by recruiting histone deacetylases (HDACs) to the estrogen response element containing target gene chromatin in breast cancer cells. Using a yeast two-hybrid screen with the MTA1 C-terminal domain as bait, we identified MAT1 (menage a trois 1) as an MTA1-binding protein. MAT1 is an assembly/targeting factor for cyclin-dependent kinase-activating kinase (CAK), which has been shown to functionally interact with general transcriptional factor TFIIH, a known inducer of ER transactivation. We show that estrogen signaling promotes nuclear translocation of MAT1 and that MTA1 interacts with MAT1 both in vitro and in vivo. MAT1 binds to the C-terminal 389-441 amino acids GATA domain and N-terminal 1-164 amino acids bromo-domain of MTA1, whereas MTA1 binds to the N-terminal ring finger domain of the MAT1. In addition, MAT1 interacts with the activation function 2 domain of ER and colocalizes with ER in activated cells. MTA1 deregulation in breast cancer cells led to its interactions with the CAK complex components, ER, and HDAC2. Accordingly, MTA1 inhibited CAK stimulation of ER transactivation that was partially relieved by HDAC inhibitor trichostatin A, suggesting that MTA1 might inhibit CAK-induced transactivation function of ER by recruiting HDAC. Furthermore, MTA1 overexpression inhibited the ability of CAK complex to phosphorylate ER. Together, these findings identified MAT1 as a target of MTA1 and provided new evidence to suggest that the transactivation functions of ER might be influenced by the regulatory interactions between CAK and MTA1 in breast cancer cells. PMID- 12527757 TI - Promoter escape by RNA polymerase II. Downstream promoter DNA is required during multiple steps of early transcription. AB - Recent evidence, obtained in a reconstituted RNA polymerase II transcription system, indicated that the promoter escape stage of transcription requires template DNA located downstream of the elongating polymerase. In the absence of downstream DNA, very early elongation complexes are unable to synthesize transcripts longer than approximately 10-14 nucleotides. In contrast, once transcripts longer than approximately 15 nucleotides have been synthesized, an extended region of downstream DNA is no longer required (Dvir, A., Tan, S., Conaway, J. W., and Conaway, R. C. (1997) J. Biol. Chem. 272, 28175-28178). In this work, we sought to define precisely when, during the synthesis of the first 10-15 phosphodiester bonds, downstream DNA is required. We report that, for complete promoter escape, downstream DNA extending to position 40/42 is required. The polymerase can be forced to arrest at several points prior to the completion of promoter escape by removing downstream DNA proximally to positions 40/42. The positions at which the polymerase arrests appear to be determined by the length of available downstream DNA, with arrest occurring at a relatively fixed position of approximately 28 nucleotides to the distal end of the template. A similar requirement is observed for transcription initiation, i.e. the formation of the first phosphodiester bond of nascent transcripts. In addition, we show that the requirement for a downstream region is independent of downstream DNA sequence, suggesting that the requirement reflects a general mechanism. Taken together, our results indicate (i) that downstream DNA is required continuously through the synthesis of the first 14-15 phosphodiester bonds of nascent transcripts, and (ii) that a major conformational change in the transcription complex likely occurs only after the completion of promoter escape. PMID- 12527758 TI - Glucose-mediated phosphorylation converts the transcription factor Rgt1 from a repressor to an activator. AB - Glucose, the most abundant carbon and energy source, regulates the expression of genes required for its own efficient metabolism. In the yeast Saccharomyces cerevisiae, glucose induces the expression of the hexose transporter (HXT) genes by modulating the activity of the transcription factor Rgt1 that functions as a repressor when glucose is absent. However, in the presence of high concentrations of glucose, Rgt1 is converted from a repressor to an activator and is required for maximal induction of HXT1 gene expression. We report that Rgt1 binds to the HXT1 promoter only in the absence of glucose, suggesting that Rgt1 increases HXT1 gene expression at high levels of glucose by an indirect mechanism. It is likely that Rgt1 stimulates the expression of an activator of the HXT1 gene at high concentrations of glucose. In addition, we demonstrate that Rgt1 becomes hyperphosphorylated in response to high glucose levels and that this phosphorylation event is required for Rgt1 to activate transcription. Furthermore, Rgt1 lacks the glucose-mediated phosphorylation in the snf3 rgt2 and grr1 mutants, which are defective in glucose induction of HXT gene expression. In these mutants, Rgt1 behaves as a constitutive repressor independent of the carbon source. We conclude that phosphorylation of Rgt1 in response to glucose is required to abolish the Rgt1-mediated repression of the HXT genes and to convert Rgt1 from a transcriptional repressor to an activator. PMID- 12527760 TI - Structural classification of zinc fingers: survey and summary. AB - Zinc fingers are small protein domains in which zinc plays a structural role contributing to the stability of the domain. Zinc fingers are structurally diverse and are present among proteins that perform a broad range of functions in various cellular processes, such as replication and repair, transcription and translation, metabolism and signaling, cell proliferation and apoptosis. Zinc fingers typically function as interaction modules and bind to a wide variety of compounds, such as nucleic acids, proteins and small molecules. Here we present a comprehensive classification of zinc finger spatial structures. We find that each available zinc finger structure can be placed into one of eight fold groups that we define based on the structural properties in the vicinity of the zinc-binding site. Three of these fold groups comprise the majority of zinc fingers, namely, C2H2-like finger, treble clef finger and the zinc ribbon. Evolutionary relatedness of proteins within fold groups is not implied, but each group is divided into families of potential homologs. We compare our classification to existing groupings of zinc fingers and find that we define more encompassing fold groups, which bring together proteins whose similarities have previously remained unappreciated. We analyze functional properties of different zinc fingers and overlay them onto our classification. The classification helps in understanding the relationship between the structure, function and evolutionary history of these domains. The results are available as an online database of zinc finger structures. PMID- 12527759 TI - Mutagenic potentials of damaged nucleic acids produced by reactive oxygen/nitrogen species: approaches using synthetic oligonucleotides and nucleotides: survey and summary. AB - DNA and DNA precursors (deoxyribonucleotides) suffer damage by reactive oxygen/nitrogen species. They are important mutagens for organisms, due to their endogenous formation. Damaged DNA and nucleotides cause alterations of the genetic information by the mispairing properties of the damaged bases, such as 8 hydroxyguanine (7,8-dihydro-8-oxoguanine) and 2-hydroxyadenine. Here, the author reviews the mutagenic potentials of damaged bases in DNA and of damaged DNA precursors formed by reactive oxygen/nitrogen species, focusing on the results obtained with synthetic oligonucleotides and 2'-deoxyribonucleoside 5' triphosphates. PMID- 12527761 TI - Mutational analysis of the Chlamydia trachomatis dnaK promoter defines the optimal -35 promoter element. AB - A long-standing question in the biology of the intracellular bacterium, Chlamydia, has been the structure of the promoter recognized by its RNA polymerase. The 'RNA polymerase sigma subunit paradox' refers to the difficulty reconciling the conservation between the RNA polymerases of Chlamydia and Escherichia coli, especially at the level of the promoter-recognition sigma subunit, with the general lack of homology between chlamydial promoters and the E.coli sigma(70) consensus promoter. While the -10 promoter element appears to be conserved between Chlamydia and E.coli, the structure of the chlamydial -35 promoter element has not been defined. We have investigated the structure of the 35 element of the Chlamydia trachomatis dnaK promoter by measuring the effects of single base pair substitutions on in vitro promoter activity. Most substitutions produced large decreases in promoter activity, which allowed us to define the optimal -35 sequence in the context of the dnaK promoter. We found that the optimal chlamydial -35 promoter sequence is identical to the E.coli sigma(70) consensus -35 promoter element (TTGACA). These results indicate that the optimal promoter specificities of the major form of chlamydial RNA polymerase and E.coli sigma(70) RNA polymerase are in fact highly conserved. A further implication of our results is that many chlamydial promoters have a suboptimal promoter structure. We hypothesize that these chlamydial promoters are intrinsically weak promoters that can be regulated during the chlamydial developmental cycle by additional transcription factors. PMID- 12527762 TI - Cellular uptake of Antennapedia Penetratin peptides is a two-step process in which phase transfer precedes a tryptophan-dependent translocation. AB - Several homeodomains and homeodomain-containing proteins enter live cells through a receptor- and energy-independent mechanism. Translocation through biological membranes is conferred by the third alpha-helix of the homeodomain, also known as Penetratin. Biophysical studies demonstrate that entry of Penetratin into cells requires its binding to surface lipids but that binding and translocation are differentially affected by modifications of some physico-chemical properties of the peptide, like helical amphipathicity or net charge. This suggests that the plasma membrane lipid composition affects the internalization of Penetratin and that internalization requires both lipid binding and other specific properties. Using a phase transfer assay, it is shown that negatively charged lipids promote the transfer of Penetratin from a hydrophilic into a hydrophobic environment, probably through charge neutralization. Accordingly, transfer into a hydrophobic milieu can also be obtained in the absence of negatively charged lipids, by the addition of DNA oligonucleotides. Strikingly, phase transfer by charge neutralization was also observed with a variant peptide of same charge and hydrophobicity in which the tryptophan at position 6 was replaced by a phenylalanine. However, Penetratin, but not its mutant version, is internalized by live cells. This underscores that charge neutralization and phase transfer represent only a first step in the internalization process and that further crossing of a biological membrane necessitates the critical tryptophan residue at position 6. PMID- 12527763 TI - Basal transcriptional regulation of human damage-specific DNA-binding protein genes DDB1 and DDB2 by Sp1, E2F, N-myc and NF1 elements. AB - The human DDB1 and DDB2 genes encode the 127 and 48 kDa subunits, respectively, of the damage-specific DNA-binding protein (DDB). Mutations in the DDB2 gene have been correlated with the hereditary disease xeroderma pigmentosum group E. We have investigated the proximal promoters of the DDB genes, both of which are G/C rich and do not contain a TATA box. Transient expression analysis in HeLa cells using a luciferase reporter system indicated the presence of core promoters located within 292 bp (DDB1) and 220 bp (DDB2) upstream of the putative transcription initiation sites. Both core promoters contain multiple active Sp1 sites, with those of DDB1 at -123 to -115 and of DDB2 at -29 to -22 being critical determinants of promoter activity. In addition, an N-myc site at -56 to 51 for DDB1 is an essential transcription element, and mutations in a DDB1 NF-1 site at -104 to -92, a DDB2 NF-1 site at -68 to -56 and a DDB2 E2F site at +36 to +43 also reduce promoter activity. Taken together, these results suggest a regulation of basal transcription typical of cell cycle-regulated genes, and therefore support conjectures that the DDB heterodimer and/or its subunits have functions other than direct involvement in DNA repair. PMID- 12527764 TI - Identification and functional characterization of a new member of the human Mcm protein family: hMcm8. AB - The six minichromosome maintenance proteins (Mcm2-7) are required for both the initiation and elongation of chromosomal DNA, ensuring that DNA replication takes place once, and only once, during the S phase. Here we report on the cloning of a new human Mcm gene (hMcm8) and on characterisation of its protein product. The hMcm8 gene contains the central Mcm domain conserved in the Mcm2-7 gene family, and is expressed in a range of cell lines and human tissues. hMcm8 mRNA accumulates during G(1)/S phase, while hMcm8 protein is detectable throughout the cell cycle. Immunoprecipitation-based studies did not reveal any participation of hMcm8 in the Mcm3/5 and Mcm2/4/6/7 subcomplexes. hMcm8 localises to the nucleus, although it is devoid of a nuclear localisation signal, suggesting that it binds to a nuclear protein. In the nucleus, the hMcm8 structure-bound fraction is detectable in S, but not in G(2)/M, phase, as for hMcm3. However, unlike hMcm3, the hMcm8 structure-bound fraction is not detectable in G(1) phase. Overall, our data identify a new Mcm protein, which does not form part of the Mcm2-7 complex and which is only structure-bound during S phase, thus suggesting its specific role in DNA replication. PMID- 12527765 TI - Site-directed mutagenesis analysis of the structural interaction of the single strand-break repair protein, X-ray cross-complementing group 1, with DNA polymerase beta. AB - Human X-ray cross-complementing group 1 (XRCC1) is a single-strand DNA break repair protein which forms a base excision repair (BER) complex with DNA polymerase beta (beta-Pol). Here we report a site- directed mutational analysis in which 16 mutated versions of the XRCC1 N-terminal domain (XRCC1-NTD) were constructed on the basis of previous NMR results that had implicated the proximity of various surface residues to beta-Pol. Mutant proteins defective in XRCC1-NTD interaction with beta-Pol and with a beta-Pol-gapped DNA complex were determined by gel filtration chromatography and a gel mobility shift assay. The interaction surface determined from the mutated residues was found to encompass beta-strand D and E of the five-stranded beta-sheet (betaABGDE) and the protruding alpha2 helix of the XRCC1-NTD. Mutations that included F67A (betaD), E69K (betaD), V86R (betaE) on the five-stranded beta-sheet and deletion of the alpha2 helix, but not mutations within alpha2, abolished binding of the XRCC1-NTD to beta-Pol. A Y136A mutant abolished beta-Pol binding, and a R109S mutant reduced beta-Pol binding. E98K, E98A, N104A, Y136A, R109S, K129E, F142A, R31A/K32A/R34A and delta-helix-2 mutants displayed temperature dependent solubility. These findings confirm the importance of the alpha2 helix and the betaD and betaE strands of XRCC1-NTD to the energetics of beta-Pol binding. Establishing the direct contacts in the beta-Pol XRCC1 complex is a critical step in understanding how XRCC1 fulfills its numerous functions in DNA BER. PMID- 12527766 TI - Tolerance for mutations and chemical modifications in a siRNA. AB - Short interfering RNA (siRNA), the active agent of RNA interference, shows promise of becoming a valuable tool in both basic and clinical research. We explore the tolerance to mutations and chemical modifications in various parts of the two 21-nt strands of a siRNA targeting the blood clotting initiator Tissue Factor. The mutations were G/C transversions. The chemical modifications were 2' O-methylation, 2'-O-allylation and phosphorothioates. We found that siRNA generally tolerated mutations in the 5' end, while the 3' end exhibited low tolerance. This observation may facilitate the design of siRNA for specific targeting of transcripts containing single nucleotide polymorphisms. We further demonstrate that in our system the single antisense strand of the wild-type siRNA is almost as effective as the siRNA duplex, while the corresponding methylated M2+4 version of the antisense had reduced activity. Most of the chemically modified versions tested had near-wild-type initial activity, while the long-term activity was increased for certain siRNA species. Our results may improve the design of siRNAs for in vivo experiments. PMID- 12527767 TI - The pathogenic U3271C human mitochondrial tRNA(Leu(UUR)) mutation disrupts a fragile anticodon stem. AB - The U3271C mutation affecting the human mitochondrial transfer RNA(Leu(UUR)) (hs mt tRNA) is correlated with diabetes and mitochondrial encephalopathies. We have explored the relationship between the structural effects of this mutation and its impact on function using chemical probing experiments and in vitro aminoacylation assays to investigate a series of tRNA constructs. Chemical probing experiments indicate that the U3271C substitution, which replaces an AU pair with a CA mispair, significantly destabilizes the anticodon stem. The introduction of a compensatory A3261G mutation reintroduces base pairing at this site and restores the structure of this domain. In fact, the anticodon stem of the A3261G/U3271C mutant appears more structured than wild-type (WT) hs mt tRNA(Leu(UUR)), indicating that the entirely AU stem of the native tRNA is intrinsically weak. The results of the chemical probing experiments are mirrored in the aminoacylation activities of the mutants. The U3271C substitution decreases aminoacylation reactivity relative to the WT tRNA due to an increase in K(m) for the pathogenic mutant. The binding defect is a direct result of the structural disruption caused by the pathogenic mutation, as the introduction of the stabilizing compensatory mutation restores aminoacylation activity. Other examples of functional defects associated with the disruption of weak domains in hs mt tRNAs have been reported, indicating that the effects of pathogenic mutations may be amplified by the fragile structures that are characteristic of this class of tRNAs. PMID- 12527768 TI - Human RNase 7: a new cationic ribonuclease of the RNase A superfamily. AB - Here we report on the expression and function of RNase 7, one of the final RNase A superfamily ribonucleases identified in the human genome sequence. The human RNase 7 gene is expressed in various somatic tissues including the liver, kidney, skeletal muscle and heart. Recombinant RNase 7 is ribonucleolytically active against yeast tRNA, as expected from the presence of eight conserved cysteines and the catalytic histidine-lysine- histidine triad which are signature motifs of this superfamily. The protein is atypically cationic with an isoelectric point (pI) of 10.5. Expression of recombinant RNase 7 in Escherichia coli completely inhibits the growth of the host bacteria, similar to what has been observed for the cationic RNase, eosinophil cationic protein (ECP/RNase 3, pI 11.4). An in vitro assay demonstrates dose-dependent cytotoxicity of RNase 7 against bacteria E.coli, Pseudomonas aeruginosa and Staphylococcus aureus. While RNase 7 and ECP/RNase 3 are both cationic and share this particular aspect of functional similarity, their protein sequence identity is only 40%. Of particular interest, ECP/RNase 3's cationicity is based on an (over)abundance of arginine residues, whereas RNase 7 includes an excess of lysine. This difference, in conjunction with the independent origins and different expression patterns, suggests that RNase 7 and ECP/RNase 3 may have been recruited to target different pathogens in vivo, if their physiological functions are indeed host defenses. PMID- 12527769 TI - Identification of a cis-acting element required for shunt-mediated translational initiation of the Sendai virus Y proteins. AB - Shunting is a mechanism that permits translational initiation at internal codons positioned in proximity to a ribosome acceptor sequence. Sendai virus exploits shunting to express a series of proteins that initiate at the fourth and fifth start sites on the P/C mRNA (namely, the Y1 and Y2 proteins, respectively). Shunt mediated initiation at these sites is codon independent. In an attempt to characterise the acceptor site, an extensive deletion analysis was performed spanning the entire C ORF. Only mutants flanking the Y1/Y2 start sites exhibited altered shunt phenotypes. Some of these significantly enhanced shunting efficiency to the point where the Y1/Y2 proteins were the major translational products of the mRNA. Additionally, removal of a short region just downstream of the Y2 start codon (referred to as Delta10) ablated all Y protein initiation via shunting but had no effect on Y expression when the AUG codons were viewed by a scanning ribosome. Point mutations introduced into this Delta10 sequence severely perturbed shunt-mediated initiation. We also provide evidence that changes in this region of the P/C mRNA may be used to modulate Y protein expression levels in different viral strains. PMID- 12527770 TI - Putative subunits of the maize origin of replication recognition complex ZmORC1 ZmORC5. AB - The finding in animal species of complexes homologous to the products of six Saccharomyces cerevisiae genes, origin of replication recognition complex (ORC), has suggested that ORC-related mechanisms have been conserved in all eukaryotes. In plants, however, the only cloned putative homologs of ORC subunits are the Arabidopsis ORC2 and the rice ORC1. Homologs of other subunits of plant origin have not been cloned and characterized. A striking observation was the absence from the Arabidopsis genome of an obvious candidate gene-homolog of ORC4. This fact raised compelling questions of whether plants, in general, and Arabidopsis, in particular, may have lost the ORC4 gene, whether ORC-homologous subunits function within a complex in plants, whether an ORC complex may form and function without an ORC4 subunit, whether a functional (but not sequence) protein homolog may have taken up the role of ORC4 in Arabidopsis, and whether lack of ORC4 is a plant feature, in general. Here, we report the first cloned and molecularly characterized five genes coding for the maize putative homologs of ORC subunits ZmORC1, ZmORC2, ZmORC3, ZmORC4 and ZmORC5. Their expression profiles in tissues with different cell-dividing activities are compatible with a role in DNA replication. Based on the potential of ORC-homologous maize proteins to bind each other in yeast, we propose a model for their possible assembly within a maize ORC. The isolation and molecular characterization of an ORC4-homologous gene from maize argues that, in its evolution, Arabidopsis may have lost the homologous ORC4 gene. PMID- 12527771 TI - Binding interactions between the core central domain of 16S rRNA and the ribosomal protein S15 determined by molecular dynamics simulations. AB - The goal of the current study is to utilize molecular dynamic (MD) simulations to investigate the dynamic behavior of 16S rRNA in the presence and absence of S15 and to identify the binding interactions between these two molecules. The simulations show that: (i) 16S rRNA remains in a highly folded structure when it is bound to S15; (ii) in the absence of S15, 16S rRNA significantly alters its conformation and transiently forms conformations that are similar to the bound structure that make it available for binding with S15; (iii) the unbound rRNA spends the majority of its time in extended conformations. The formation of the extended conformations is a result of the molecule reaching a lower electrostatic energy and the formation of the highly folded, crystal-like conformation is a result of achieving a lower solvation energy. In addition, our MD simulations show that 16S rRNA and S15 bind across the major groove of helix 22 (H22) via electrostatic interactions. The negatively charged phosphate groups of G658, U740, G741 and G742 bind to the positively charged S15 residues Lys7, Arg34 and Arg37. The current study provides a dynamic view of the binding of 16S rRNA with S15. PMID- 12527772 TI - Polypyrimidine tract binding protein and poly r(C) binding protein 1 interact with the BAG-1 IRES and stimulate its activity in vitro and in vivo. AB - The 5'-untranslated region of Bag-1 mRNA contains an internal ribosome entry segment (IRES) and the translation of Bag-1 protein can be initiated by both cap dependent and cap-independent mechanisms. In general, cellular IRESs require non canonical trans-acting factors for their activity, however, very few of the proteins that act on cellular IRESs have been identified. Proteins that interact with viral IRESs have also been shown to stimulate the activity of cellular IRESs and therefore the ability of a range of known viral trans-acting factors to stimulate the Bag-1 IRES was tested. Two proteins, poly r(C) binding protein 1 (PCBP1) and polypyrimidine tract binding protein (PTB), were found to increase the activity of the Bag-1 IRES in vitro and in vivo. The regions of the Bag-1 IRES RNA to which they bind have been determined, and it was shown that PCBP1 binds to a short 66 nt section of RNA, whilst PTB interacts with a number of sites over a larger area. The minimum section of the RNA that still retained activity was determined and both PCBP1 and PTB interacted with this region suggesting that these proteins are essential for Bag-1 IRES function. PMID- 12527773 TI - Putative proteins related to group II intron reverse transcriptase/maturases are encoded by nuclear genes in higher plants. AB - The Arabidopsis thaliana nuclear genome sequence revealed several open reading frames encoding proteins related to group II intron-encoded reverse transcriptase/maturases. Here, we show via sequence alignments that at least four such open reading frames are conserved in the nuclear genomes of A.thaliana and Oryza sativa (rice) and that they encode putative proteins belonging to two different classes (nMat-1 and nMat-2), neither of which is associated with a group II intron RNA structure. The two nMat-1 proteins have reverse transcriptase, maturase and DNA endonuclease domains characteristic of canonical group II intron-encoded proteins, while the two nMat-2 proteins have reverse transcriptase and maturase domains linked to a novel C-terminal domain. Although some nMat proteins have mutations expected to inactivate intron mobility functions, all could potentially retain the RNA splicing function. These nuclear maturase-like proteins may be imported into organelles to function in group II intron splicing and/or they may have assumed other cellular functions. Nuclear encoded maturases could regulate organellar gene expression and may reflect a step in the evolution of mobile group II introns into spliceosomal introns. PMID- 12527774 TI - The phylogenetic diversity of eukaryotic transcription. AB - Eukaryotic transcription is a highly regulated process involving interactions between large numbers of proteins. To analyse the phylogenetic distribution of the components of this process, six crown eukaryote group genomes were queried with a reference set of transcription-associated (TA) proteins. On average, one in 10 proteins encoded by these genomes were found to be homologous to sequences in the reference set. Analysis of families identified using an accurate sequence clustering algorithm and containing both TA proteins and eukaryotic sequences showed that in two-thirds of the families the homologues originate from a single kingdom. Furthermore, in only 15% of the fungal-specific clusters are the homologues present in both budding and fission yeast, as compared with the metazoan-specific clusters where 53% of the homologues originate from two or more species. Families whose members comprise general transcription factor or RNA polymerase subunits exhibit a low degree of taxon specificity, suggesting that the transcription initiation complex is highly conserved. This contrasts with transcriptional regulator families, that are primarily taxon-specific, indicating proteins controlling gene activation exhibit considerable sequence diversity across the eukaryotic domain. PMID- 12527775 TI - Selections for constituting new RNA-protein interactions in catalytic RNP. AB - In vitro and in vivo selection techniques are developed to constitute new RNA peptide interactions. The selection strategy is designed by employing a catalytic RNP consisting of a derivative of the Tetrahymena ribozyme and an artificial RNA binding protein. An arginine-rich RNA-binding motif and its target RNA motif in the RNP are substituted with randomized sequences and used for the selection experiments. Previously unknown binding motifs are obtained and the newly established interactions have been indispensable for assembling a catalytically active RNP. The method employed in this study is useful for making customized self-splicing intron RNAs whose activity is regulated by protein cofactors. PMID- 12527777 TI - Finding weak similarities between proteins by sequence profile comparison. AB - To improve the recognition of weak similarities between proteins a method of aligning two sequence profiles is proposed. It is shown that exploring the sequence space in the vicinity of the sequence with unknown properties significantly improves the performance of sequence alignment methods. Consistent with the previous observations the recognition sensitivity and alignment accuracy obtained by a profile-profile alignment method can be as much as 30% higher compared to the sequence-profile alignment method. It is demonstrated that the choice of score function and the diversity of the test profile are very important factors for achieving the maximum performance of the method, whereas the optimum range of these parameters depends on the level of similarity to be recognized. PMID- 12527776 TI - Functional studies of the PI(3)-kinase signalling pathway employing synthetic and expressed siRNA. AB - RNA interference (RNAi) is a RNA-mediated sequence-specific gene silencing mechanism. Recently, this mechanism has been used to down-regulate protein expression in mammalian cells by applying synthetic- or vector-generated small interfering RNAs (siRNAs). However, for the evaluation of this new knockdown technology, it is crucial to demonstrate biological consequences beyond protein level reduction. Here, we demonstrate that this new siRNA-based technology is suitable to analyse protein functions using the phosphatidylinositol (PI) 3 kinase signal transduction pathway as a model system. We demonstrate stable and transient siRNA-mediated knockdown of one of the PI 3-kinase catalytic subunits, p110beta, which leads to inhibition of invasive cell growth in vitro as well as in a tumour model system. Importantly, this result is consistent with loss-of function phenotypes induced by conventional RNase H-dependent antisense molecules or treatment with the PI 3-kinase inhibitor LY294002. RNAi knockdown of the downstream kinases Akt1 and Akt2 does not reduce cell growth on extracellular matrix. Our data show that synthetic siRNAs, as well as vector-based expression of siRNAs, are a powerful new tool to interfere with signal transduction processes for the elucidation of gene function in mammalian cells. PMID- 12527778 TI - Enp1, a yeast protein associated with U3 and U14 snoRNAs, is required for pre rRNA processing and 40S subunit synthesis. AB - ENP1 is an essential Saccharomyces cerevisiae gene encoding a 483 amino acid polypeptide. Enp1 protein is localized in the nucleus and concentrated in the nucleolus. An enp1-1 temperature-sensitive mutant inhibited 35S pre-rRNA early processing at sites A(0), A(1) and A(2) as shown by northern analysis of steady state levels of rRNA precursors. Pulse-chase analysis further revealed that the enp1-1 strain was defective in the synthesis of 20S pre-rRNA and hence 18S rRNA, which led to reduced formation of 40S ribosomal subunits. Co-precipitation analysis revealed that Enp1 was associated with Nop1 protein, as well as with U3 and U14 RNAs, two snoRNAs implicated in early pre-rRNA processing steps. These results suggest a direct role for Enp1 in the early steps of rRNA processing. PMID- 12527779 TI - Short hairpin type of dsRNAs that are controlled by tRNA(Val) promoter significantly induce RNAi-mediated gene silencing in the cytoplasm of human cells. AB - The post-transcriptional gene silencing in animals and plants is called RNA interference (RNAi). Guides for the sequence-specific degradation of mRNA are 21 nt small interfering RNAs (siRNAs) that are generated by Dicer-dependent cleavage from longer double-stranded RNAs (dsRNAs). To examine the relationship between the localization of dsRNA and the target cleavage of RNAi in human cells, we constructed five kinds of dsRNA expression vector that were controlled by tRNA(Val) or U6 promoter. Transcripts of tRNA-dsRNA were consistently localized in the cytoplasm and were efficiently processed by Dicer. In contrast, transcripts of tRNA-dsRNA were not processed in cells that expressed Dicer directed ribozymes. In addition, transcripts of U6-dsRNA were basically localized in the nucleus and were not significantly processed, unless the transcripts of U6 dsRNAs possessed a microRNA-based loop motif: in the latter case, U6-dsRNAs with a microRNA-based loop were transported to the cytoplasm and were effectively processed. More over, tRNA-dsRNA directed against a mutant k-ras transcript cleaved its target mRNA efficiently in assays of RNAi not only in vitro with a cytoplasmic extract but also in vivo. Therefore, it appears that RNAi in human cells occur in the cytoplasm. Importantly, the same tRNA-dsRNA did not affect the degradation of the normal k-ras mRNA in vitro and in vivo. Our tRNA-dsRNA technology should be a powerful tool for studies of the mechanism of RNAi and the functions of various genes in mammalian cells with potential utility as a therapeutic agent. PMID- 12527780 TI - Fluoride-cleavable biotinylation phosphoramidite for 5'-end-labeling and affinity purification of synthetic oligonucleotides. AB - A fluoride-cleavable phosphoramidite for biotinylation was designed, synthesized and coupled efficiently to the 5'-end of DNA on an automatic synthesizer. The diisopropylsilyl acetal functionality was used to link the biotin moiety through a tertiary hydroxide group to the 5'-end of DNA. This linkage proved to be completely stable under certain post-synthetic DNA cleavage/deprotection conditions [0.05 M K(2)CO(3) in MeOH, room temperature, 24 h and MeNH(2) (approximately 40%)/NH(4)OH (approximately 29%), 1:1 v/v, 65 degrees C, 30 min] while it can be readily broken by fluoride ion, releasing unmodified DNA. To demonstrate the use of this DNA biotinylation method, we applied this method in affinity purification of synthetic DNA. As revealed by HPLC analysis, biotinylated full-length DNA can be efficiently attached to NeutrAvidin coated microspheres, and failure sequences can be readily removed. Subsequent treatment of the microspheres with pyridine/HF released high quality full-length unmodified DNA in good yield. PMID- 12527781 TI - The complete nucleotide sequence of the hornwort (Anthoceros formosae) chloroplast genome: insight into the earliest land plants. AB - It is generally believed that bryophytes are the earliest land plants. However, the phylogenetic relationships among bryophytes, including mosses, liverworts and hornworts, are not clearly resolved. To obtain more information on the earliest land plants, we determined the complete nucleotide sequence of the chloroplast genome from the hornwort Anthoceros formosae. The circular double-stranded DNA of 161 162 bp is the largest genome ever reported among land plant chloroplasts. It contains 76 protein, 32 tRNA and 4 rRNA genes and 10 open reading frames (ORFs), which are identical with the chloroplast genome of the other green plants analyzed. The major difference is a larger inverted repeat than that of the liverwort Marchantia, Anthoceros contains an excess of ndhB and rps7 genes and the 3' exon of rps12. The genes matK and rps15, commonly found in the chloroplast genomes of land plants, are pseudogenes. The intron of rrn23 is the first finding in the known chloroplast genomes of land plants. A striking feature of the hornwort chloroplast is that more than half of the protein-coding genes have nonsense codons, which are converted into sense codons by RNA editing. Maximum likelihood (ML) analysis, based on 11 518 amino acid sites of 52 proteins encoded in the chloroplast genomes of the green plants, placed liverworts as the sister to all other land plants. PMID- 12527782 TI - Comparison of the capacity of different viral internal ribosome entry segments to direct translation initiation in poly(A)-dependent reticulocyte lysates. AB - Polyadenylation stimulates translation of capped eukaryotic mRNAs and those carrying picornaviral internal ribosome entry segments (IRESes) in vivo. Rabbit reticulocyte lysates (RRL) reproduce poly(A)-mediated translation stimulation in vitro after partial depletion of ribosomes and ribosome-associated factors. Here, we have evaluated the effects of varying different parameters (extent of extract depletion, cleavage of eIF4G, concentrations of KCl, MgCl(2) and programming mRNA) on IRES-driven translation efficiency and poly(A)-dependency in ribosome depleted RRL. For comparison, the study included a standard capped, polyadenylated mRNA. Dramatic differences were observed in the abilities of the different IRESes to direct translation in ribosome-depleted extracts. While the hepatitis A virus IRES was incapable of driving translation in physiological conditions in depleted RRL, mRNAs carrying the foot-and-mouth disease virus and hepatitis C virus IRESes were translated significantly better than a standard cellular mRNA in the same conditions. Indeed, the capacities of these IRESes to direct translation in ribosome-depleted RRL were similar to those reported previously in certain cell lines. Both the abilities of the IRESes to drive translation and their individual salt optima in ribosome-depleted extracts suggest that these elements have dramatically different affinities for some component(s) of the canonical translation machinery. Finally, using poliovirus as an example, we show that the ribosome-depleted system is well suited to the study of the translational capacity of naturally occurring IRES variants. PMID- 12527783 TI - Antisense oligonucleotides targeted to the domain IIId of the hepatitis C virus IRES compete with 40S ribosomal subunit binding and prevent in vitro translation. AB - Initiation of protein synthesis on the hepatitis C virus (HCV) mRNA involves a structured element corresponding to the 5' untranslated region and constituting an internal ribosome entry site (IRES). The domain IIId of the HCV IRES, an imperfect RNA hairpin extending from nucleotides 253 to 279 of the viral mRNA, has been shown to be essential for translation and for the binding of the 40S ribosomal subunit. We investigated the properties of a series of antisense 2'-O methyloligoribonucleotides targeted to various portions of the domain IIId. Several oligomers, 14-17 nt in length, selectively inhibited in vitro translation of a bicistronic RNA construct in rabbit reticulocyte lysate with IC(50)s <10 nM. The effect was restricted to the second cistron (the Renilla luciferase) located downstream of the HCV IRES; no effect was observed on the expression of the first cistron (the firefly luciferase) which was translated in a cap-dependent manner. Moreover, antisense 2'-O-methyloligoribonucleotides specifically competed with the 40S ribosomal subunit for binding to the IRES RNA in a filter- retention assay. The antisense efficiency of the oligonucleotides was nicely correlated to their affinity for the IIId subdomain and to their ability to displace 40S ribosomal subunit, making this process a likely explanation for in vitro inhibition of HCV-IRES-dependent translation. PMID- 12527784 TI - Esp1396I restriction-modification system: structural organization and mode of regulation. AB - Esp1396I restriction-modification (RM) system recognizes an interrupted palindromic DNA sequence 5'-CCA(N)(5)TGG-3'. The Esp1396I RM system was found to reside on pEsp1396, a 5.6 kb plasmid naturally occurring in Enterobacter sp. strain RFL1396. The nucleotide sequence of the entire 5622 bp pEsp1396 plasmid was determined on both strands. Identified genes for DNA methyltransferase (esp1396IM) and restriction endonuclease (esp1396IR) are transcribed convergently. The restriction endonuclease gene is preceded by the small ORF (esp1396IC) that possesses a strong helix-turn-helix motif and resembles regulatory proteins found in PvuII, BamHI and few other RM systems. Gene regulation studies revealed that C.Esp1396I acts as both a repressor of methylase expression and an activator of regulatory protein and restriction endonuclease expression. Our data indicate that C protein from Esp1396I RM system activates the expression of the Enase gene, which is co-transcribed from the promoter of regulatory gene, by the mechanism of coupled translation. PMID- 12527785 TI - Prioritized selection of oligodeoxyribonucleotide probes for efficient hybridization to RNA transcripts. AB - Only a small fraction of short oligonucleotide probes bind efficiently to complementary segments in long RNA transcripts. Technologies such as array-based transcript profiling and antisense control of gene expression would benefit greatly from a method for predicting probes that bind well to a given target RNA. To develop an algorithm for prioritizing selection of probes, we have analyzed predicted thermodynamic parameters for the binding of several large sets of probes to complementary RNA transcripts. The binding of five of these sets of probes to their RNA targets has been reported by others. In addition, we have used a method for light-directed synthesis of oligonucleotide arrays that we developed to generate two new arrays of surface-bound probes and measured the binding of these probes to their RNA targets. We considered predicted free energies for intramolecular base pairing of the oligonucleotide and its RNA target as well as the predicted free energy of intermolecular hybridization of probe and target. We find that a reliable predictor of probes that will hybridize significantly with their targeted transcripts is the predicted free energy of hybridization minus the predicted free energy for intramolecular folding of the probe. PMID- 12527786 TI - A comparison of three fission yeast mitochondrial genomes. AB - The fission yeasts are members of the fungal order Schizosaccharomycetales, a candidate deep-diverging group within Ascomycota. Although a great deal of molecular information is available from Schizosaccharomyces pombe, a model eukaryote, very little is available from other members of this group. In order to better characterize mitochondrial genome evolution in this fungal lineage, the mitochondrial DNA (mtDNA) of two additional fission yeasts, Schizosaccharomyces octosporus and Schizosaccharomyces japonicus var. japonicus, was sequenced. Whereas the mtDNA of S.pombe is only 19 431 bp, the mtDNA of S.octosporus is 44 227 bp, and that of S.japonicus var. japonicus is over 80 kb. The size variation of these mtDNAs is due largely to non-coding regions. The gene content in the latter two mtDNAs is almost identical to that of the completely sequenced S.pombe mtDNA, which encodes 25 tRNA species, the large and small mitochondrial ribosomal RNAs (rnl and rns), the RNA component of mitochondrial RNaseP (rnpB), mitochondrial small subunit ribosomal protein 3 (rps3), cytochrome oxidase subunits 1, 2 and 3 (cox1, cox2 and cox3) and ATP-synthase subunits 6, 8 and 9 (atp6, atp8 and atp9). However, trnI2(cau) (C modified to lysidine) is absent in the S.octosporus mtDNA, as are corresponding ATA codons in its protein-coding genes, and rps3 and rnpB are not found in the mtDNA of S.japonicus var. japonicus. The mtDNA of S.octosporus contains five double hairpin elements, the first report of these elements in an ascomycete. This study provides further evidence in favor of the mobility of these elements, and supports their role in mitochondrial genome rearrangement. The results of our phylogenetic analysis support the monophyly of the Schizosaccharomycetales, but question their grouping within the Archiascomycota. PMID- 12527787 TI - Characterization of fragmented mitochondrial ribosomal RNAs of the colorless green alga Polytomella parva. AB - We have identified previously in mitochondrial DNA of the colorless, chlorophycean, green algal taxon, Polytomella parva, potential coding regions for four small subunit (SSU) and eight large subunit (LSU) rRNA fragments. In this study with P.parva, we isolated RNA from a mitochondrial-enriched preparation, characterized the 12 mitochondrial rRNA transcripts by either northern blot analysis or chemical sequencing and performed secondary structure modeling of the SSU and LSU rRNA sequences. The results show the following features about the mitochondrial SSU and LSU rRNAs of P.parva: (i) they are considerably shorter than their homologs from other green algae, although the main domains typical of conventional rRNAs are conserved; (ii) the rRNA fragmentation pattern is most similar to that of Chlamydomonas reinhardtii among green algae that have been characterized; (iii) three nucleotides are missing from the normally highly conserved GTPase center of the LSU rRNA; and (iv) post-transcriptional modification of the 3'-terminal region of the SSU rRNA is unusual in that it has the 'eubacterial' 3-methyluridine (corresponding to m(3)U at Escherichia coli 16S rRNA position 1498) but lacks the more highly conserved modifications at two adjacent A residues (corresponding to N(6),N(6)-dimethyladenosine at E.coli 16S rRNA positions 1518 and 1519). This is the first report of the characterization by direct sequencing of fragmented mitochondrial rRNAs from a green alga. PMID- 12527788 TI - Fingerprinting of prokaryotic 16S rRNA genes using oligodeoxyribonucleotide microarrays and virtual hybridization. AB - An oligonucleotide microarray hybridization system to differentiate microbial species was designed and tested. Seven microbial species were studied, including one Bacillus and six Pseudomonas strains. DNA sequences near the 5' end of 16S rRNA genes were aligned and two contiguous regions of high variability, flanked by highly conserved sequences, were found. The conserved sequences were used to design PCR primers which efficiently amplified these polymorphic regions from all seven species. The amplicon sequences were used to design 88 9mer hybridization probes which were arrayed onto glass slides. Single-stranded, fluorescence-tagged PCR products were hybridized to the microarrays at 15 degrees C. The experimental results were compared with the DeltaG(0) values for all matched and mismatched duplexes possible between the synthetic probes and the 16S target sequences of the seven test species, calculated using a 'virtual hybridization' software program. Although the observed hybridization patterns differed significantly from patterns predicted solely on the basis of perfect sequence matches, a unique hybridization fingerprint was obtained for each of the species, including closely related Pseudomonas species, and there was a reasonable correlation between the intensity of observed hybridization signals and the calculated DeltaG(0) values. The results suggest that both perfect and mismatched pairings can contribute to microbial identification by hybridization fingerprinting. PMID- 12527789 TI - Inferring functional constraints and divergence in protein families using 3D mapping of phylogenetic information. AB - Comparative sequence analysis has been used to study specific questions about the structure and function of proteins for many years. Here we propose a knowledge based framework in which the maximum likelihood rate of evolution is used to quantify the level of constraint on the identity of a site. We demonstrate that site-rate mapping on 3D structures using datasets of rhodopsin-like G-protein receptors and alpha- and beta-tubulins provides an excellent tool for pinpointing the functional features shared between orthologous and paralogous proteins. In addition, functional divergence within protein families can be inferred by examining the differences in the site rates, the differences in the chemical properties of the side chains or amino acid usage between aligned sites. Two novel analytical methods are introduced to characterize rate- independent functional divergence. These are tested using a dataset of two classes of HMG-CoA reductases for which only one class can perform both the forward and reverse reaction. We show that functionally divergent sites occur in a cluster of sites interacting with the catalytic residues and that this information should facilitate the design of experimental strategies to directly test functional properties of residues. PMID- 12527790 TI - Specificity assessment from fractionation experiments (SAFE): a novel method to evaluate microarray probe specificity based on hybridisation stringencies. AB - The cDNA-chip technology is a highly versatile tool for the comprehensive analysis of gene expression at the transcript level. Although it has been applied successfully in expression profiling projects, there is an ongoing dispute concerning the quality of such expression data. The latter critically depends on the specificity of hybridisation. SAFE (specificity assessment from fractionation experiments) is a novel method to discriminate between non- specific cross hybridisation and specific signals. We applied in situ fractionation of hybridised target on DNA-chips by means of repeated washes with increasing stringencies. Different fractions of hybridised target are washed off at defined stringencies and the collected fluorescence intensity data at each step comprise the fractionation curve. Based on characteristic features of the fractionation curve, unreliable data can be filtered and eliminated from subsequent analyses. The approach described here provides a novel experimental tool to identify probes that produce specific hybridisation signals in DNA-chip expression profiling approaches. The iterative use of the SAFE procedure will result in increasingly reliable sets of probes for microarray experiments and significantly improve the overall efficiency and reliability of RNA expression profiling data from DNA-chip experiments. PMID- 12527791 TI - Capillary electrophoretic analysis of genomic DNA methylation levels. AB - Changes in DNA methylation have been found in the large majority of tumors. This phenomenon includes both genome-wide hypomethylation and gene- specific hypermethylation. However, the clinical relevance of either mechanism has remained contentious. In order to determine DNA methylation levels from a large number of clinical samples, we have established a method for accurate high throughput quantification of 5-methylcytosine in genomic DNA. Our protocol requires a small amount (<1 micro g) of DNA that is enzymatically hydrolyzed to single nucleotides. Single nucleotides are then derivatized with a fluorescent marker and separated by capillary electrophoresis. After calibration of the method, we have determined cytosine methylation levels from tumor samples of 81 patients that had been diagnosed with chronic lymphocytic leukemia (CLL). These patients showed a high variability in their methylation levels with a general trend towards hypomethylation. Because of its high accuracy and throughput our method will be useful in determining the role of genomic DNA methylation levels in tumorigenesis. PMID- 12527792 TI - SYBR Green real-time telomeric repeat amplification protocol for the rapid quantification of telomerase activity. AB - The sensitive telomeric repeat amplification protocol (TRAP) permits telomerase detection in mammalian cell and tissue extracts with very low telomerase activity levels. Unfortunately, conventional TRAP assays require complex post amplification procedures, such as polyacrylamide gel electrophoresis and densitometry, to measure telomerase products. Therefore, a real-time quantitative TRAP assay (RQ-TRAP) was optimized in the present study and evaluated in comparison with a commercially available quantitative TRAP kit and by monitoring telomerase activity in human hepatocyte cultures, human hepatoma cell lines and telomerase reconstitution experiments. The novel real-time telomerase detection method has many advantages. Other than sample extraction and real-time cycling, no additional time-consuming steps have to be performed for telomerase quantification; reliable and linear telomerase quantification is possible down to single-cell dilutions without the interference of primer-dimer artifacts, and the costs are less. Moreover, the precision is similar to other amplification-based telomerase quantification assays and the results are comparable to data obtained with two commercially available assays. The closed-tube system reduces the risk of carryover contamination and supports high throughput. In conclusion, RQ-TRAP provides a new tool for the rapid and reliable quantification of telomerase activity. PMID- 12527793 TI - A rapid biosensor chip assay for measuring of telomerase activity using surface plasmon resonance. AB - Considerable interest has been focused on telomerase because of its potential use in assays for cancer diagnosis, and for anti-telomerase drugs as a strategy for cancer chemotherapy. A number of assays based on the polymerase chain reaction (PCR) have been developed for evaluation of telomerase activity. To overcome the disadvantages of the conventional telomerase assay [telomeric repeat amplification protocol (TRAP)] related to PCR artifacts and troublesome post-PCR procedures, we have developed a telomeric repeat elongation (TRE) assay which directly measures telomerase activity as the telomeric elongation rate by biosensor technology using surface plasmon resonance (SPR). 5'-Biotinylated oligomers containing telomeric repeats were immobilized on streptavidin pretreated dextran sensor surfaces in situ using the BIACORE apparatus. Subsequently, the oligomers associated with the telomerase extracts were elongated in the BIACORE apparatus. The rate of TRE was calculated by measuring the SPR signals. We examined elongation rates by the TRE assay in 18 cancer and three normal human fibroblast cell lines, and 12 human primary carcinomas and matching normal tissues. The elongation rates increased in a concentration- and time-dependent manner. Those of cancer cells were two to 10 times higher than fibroblast cell lines and normal tissues. Telomerase activities and its inhibitory effects of anti-telomerase agents as measured by both the TRE and TRAP assays showed a good correlation. Our assay allows precise quantitative comparison of a wide range of human cells from somatic cells to carcinoma cells. TRE assay is suitable for practical use in the assessment of telomerase activity in preclinical and clinical trials of telomerase-based therapies, because of its reproducibility, rapidity and simplicity. PMID- 12527794 TI - NotI passporting to identify species composition of complex microbial systems. AB - We describe here a new method for large-scale scanning of microbial genomes on a quantitative and qualitative basis. To achieve this aim we propose to create NotI passports: databases containing NotI tags. We demonstrated that these tags comprising 19 bp of sequence information could be successfully generated using DNA isolated from intestinal or fecal samples. Such NotI passports allow the discrimination between closely related bacterial species and even strains. This procedure for generating restriction site tagged sequences (RSTS) is called passporting and can be adapted to any other rare cutting restriction enzyme. A comparison of 1312 tags from available sequenced Escherichia coli genomes, generated with the NotI, PmeI and SbfI restriction enzymes, revealed only 219 tags that were not unique. None of these tags matched human or rodent sequences. Therefore the approach allows analysis of complex microbial mixtures such as in human gut and identification with high accuracy of a particular bacterial strain on a quantitative and qualitative basis. PMID- 12527795 TI - Improving baculovirus recombination. AB - Recombinant baculoviruses have established themselves as a favoured technology for the high-level expression of recombinant proteins. The construction of recombinant viruses, however, is a time consuming step that restricts consideration of the technology for high throughput developments. Here we use a targeted gene knockout technology to inactivate an essential viral gene that lies adjacent to the locus used for recombination. Viral DNA prepared from the knockout fails to initiate an infection unless rescued by recombination with a baculovirus transfer vector. Modified viral DNA allows 100% recombinant virus formation, obviates the need for further virus purification and offers an efficient means of mass parallel recombinant formation. PMID- 12527797 TI - Expression and characterization of functional dog flavin-containing monooxygenase 1. AB - A full-length dog (beagle) flavin-containing monooxygenase 1 (FMO1) cDNA (dFMO1) was obtained from liver by reverse transcription-polymerase chain reaction. The amino acid sequence of dFMO1 was 89% homologous to human FMO1. Using a baculovirus expression system in Sf-9 insect cells, dFMO1 was expressed to protein levels of 0.4 nmol/mg, as determined by immunoquantitation. The flavin content of the expressed enzyme was consistent with immunodetectable dFMO1 protein levels. Expressed dFMO1 catalyzed NADPH-dependent methyl p-tolyl sulfide oxidation, with K(m) and V(max) values of 98.6 microM and 63.8 nmol of S-oxide formed/min/mg of protein, respectively. By comparison, human FMO1 showed similar values of 87.1 microM (K(m)) and 51.0 nmol/min/mg (V(max)). Activity for dFMO1 showed characteristic pH dependence, with a 4.5-fold increase in S-oxidase activity as the incubation pH increased from 7.6 to 9.0. Human FMO1 also showed an increase in reaction rate with pH but a somewhat lower optimum of 8.0 to 8.4. dFMO1 also catalyzed imipramine N-oxidation, with a K(m) of 4.7 microM and a V(max) of 82.1 nmol/min/mg of protein. This enzyme displayed other characteristics of FMO enzymes, with rapid depletion of enzyme activity upon heating in the absence of NADPH. Protein levels of 74 pmol of dFMO1/mg of microsomal protein were determined for a pooled liver microsome sample, suggesting that this enzyme is a major canine hepatic monooxygenase. In conclusion, the expression and characterization of catalytically active dFMO1 will allow the role of this enzyme in the metabolism of xenobiotics to be determined. PMID- 12527796 TI - Non-nucleoside inhibitors of mitochondrial thymidine kinase (TK-2) differentially inhibit the closely related herpes simplex virus type 1 TK and Drosophila melanogaster multifunctional deoxynucleoside kinase. AB - 5'-O-Trityl derivatives of thymidine (dThd), (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU), and their acyclic analogs 1-[(Z)-4-triphenylmethoxy-2-butenyl]thymine (KIN-12) and (E)-5-(2-bromovinyl)-1-[(Z)-4-triphenylmethoxy-2-butenyl]uracil (KIN 52) have been synthesized and evaluated for their inhibitory activity against the amino acid sequence related mitochondrial dThd kinase (TK-2), herpes simplex virus type 1 (HSV-1) TK, and Drosophila melanogaster multifunctional 2' deoxynucleoside kinase (Dm-dNK). Several compounds proved markedly inhibitory to these enzymes and represent a new generation of nucleoside kinase inhibitors. KIN 52 was the most potent and selective inhibitor of TK-2 (IC(50), 1.3 microM; K(i), 0.50 microM; K(i)/K(m), 0.37) but was not inhibitory against HSV-1 TK and Dm-dNK at 100 microM. As found for the alternative substrate BVDU, the tritylated compounds competitively inhibited the three enzymes with respect to dThd. However, whereas BVDU behaved as a noncompetitive inhibitor (alternative substrate) of TK-2 and HSV-1 TK with respect to ATP as the varying substrate, the novel tritylated enzyme inhibitors emerged as reversible purely uncompetitive inhibitors of these enzymes. Computer-assisted modeling studies are in agreement with these findings. The tritylated compounds do not act as alternative substrates and they showed a type of kinetics against the nucleoside kinases different from that of BVDU. KIN-12, and particularly KIN-52, are the very first non-nucleoside specific inhibitors of TK-2 reported and may be useful for studying the physiological role of the mitochondrial TK-2 enzyme. PMID- 12527798 TI - Abrogation of the cell death response to oxidative stress by the c-Abl tyrosine kinase inhibitor STI571. AB - Normal aerobic metabolism is associated with the production of reactive oxygen species (ROS) and, consequently, the induction of apoptosis and necrosis. The cell death response to oxidative stress is thought to contribute to aging, neurological degeneration, and other disorders. ROS-induced apoptosis and necrosis involves activation of the cytoplasmic c-Abl tyrosine kinase and thereby signaling to mitochondria. Herein, we show that STI571, an inhibitor of Bcr-Abl in chronic myelogenous leukemia, blocks activation of c-Abl in the response of mouse embryo fibroblasts and human U-937 myeloid leukemia cells to hydrogen peroxide (H(2)O(2)). Immunofluorescence microscopy and subcellular fractionation studies demonstrate that STI571 decreases H(2)O(2)-induced targeting of c-Abl to mitochondria in the two cell types by 59 to 85%. The results also show that STI571 attenuates H(2)O(2)-induced loss of the mitochondrial transmembrane potential. In concert with these effects, STI571 inhibits the death response to H(2)O(2) exposure by 40 to 80% depending on the cell type. These findings indicate that inhibition of c-Abl signaling by STI571 attenuates mitochondrial dysfunction and cell death in the cellular response to oxidative stress. PMID- 12527799 TI - Opioids bind to the amino acids 84 to 118 of UDP-glucuronosyltransferase UGT2B7. AB - The UDP-glucuronosyltransferase UGT2B7 is an important human UGT isoform that catalyzes the conjugation of many endogenous and exogenous compounds, among them opioids, resulting in the formation of D-glucuronides. The binding site of the aglycone is located in the N-terminal half of the protein. Using NMR analysis, we demonstrate that the opioid binding site in UGT2B7 is within the 84 to 118 N terminal amino acids. Three maltose binding protein-UGT2B7 fusion proteins, 2B7F3 and 2B7F4 incorporating the amino acids 24 to 118 and 24 to 96 of UGT2B7, respectively, and 2B7F5 incorporating amino acids 84 to 118 of UGT2B7 were expressed in Escherichia coli and purified by affinity chromatography. NMR analysis showed that morphine was bound to the fusion protein 2B7F3 with a K(D) value similar to the K(D) values obtained for the previously produced fusion proteins, which included amino acids 24 to 180. Morphine did not bind to 2B7F4, but it did bind to 2B7F5. Both NMR 1-D spectra and NOESY experiments indicated that the 2B7F5 protein was mediating magnetization transfer within the morphine. These results allowed us to predict and model a binding site within the amino acids 96 to 101 of UGT2B7. A mutant fusion protein 2B7F3 with the substitution D99A was produced, and the NMR spectroscopy analysis of the protein supported the model. A marked reduction of morphine binding was observed when the charged aspartate was substituted with alanine. PMID- 12527800 TI - Structural requirements for imidazobenzodiazepine binding to GABA(A) receptors. AB - Several structural subclasses of ligands bind to the benzodiazepine (BZD) binding site of the GABA(A) receptor. Previous studies from this laboratory have suggested that imidazobenzodiazepines (i-BZDs, e.g., Ro 15-1788) require domains in the BZD binding site for high-affinity binding that are distinct from the requirements of classic BZDs (e.g., flunitrazepam). Here, we used systematic mutagenesis and the substituted cysteine accessibility method to map the recognition domain of i-BZDs near two residues implicated in BZD binding, gamma(2)A79 and gamma(2)T81. Both classic BZDs and i-BZDs protect cysteines substituted at gamma(2)A79 and gamma(2)T81 from covalent modification, suggesting that these ligands may occupy common volumetric spaces during binding. However, the binding of i-BZDs is more sensitive to mutations at gamma(2)A79 than classic BZDs or BZDs that lack a 3'-imidazo substituent (e.g., midazolam). The effect that gamma(2)A79 mutagenesis has on the binding affinities of a series of structurally rigid i-BZDs is related to the volume of the 3'-imidazo substituents. Furthermore, larger amino acid side chains introduced at gamma(2)A79 cause correspondingly larger decreases in the binding affinities of i BZDs with bulky 3' substituents. These data are consistent with a model in which gamma(2)A79 lines a subsite within the BZD binding pocket that accommodates the 3' substituent of i-BZDs. In agreement with our experimental data, computer assisted docking of Ro 15-4513 into a molecular model of the BZD binding site positions the 3'-imidazo substituent of Ro 15-4513 near gamma(2)A79. PMID- 12527801 TI - Basic fibroblast growth factor activates endothelial nitric-oxide synthase in CHO K1 cells via the activation of ceramide synthesis. AB - In this study, we analyzed the intracellular mechanisms leading to basic fibroblast growth factor (bFGF)-dependent production of NO in Chinese hamster ovary (CHO)-K1 cells and a possible physiological role for such an effect. bFGF induces NO production through the activation of the endothelial form of NO synthase (eNOS), causing a subsequent increase in the cGMP levels. In these cells, the activation of eNOS by bFGF is Ca(2+)- and mitogen-activated protein kinase-independent. The translocation of the enzyme from the plasma membrane, where it is located in caveolae bound to caveolin 1, to the cytosol is the crucial step for the synthesis of NO through the eNOS isoform. We demonstrate that bFGF activates a sphingomyelinase to synthesize ceramide, which, in turn, allows the dissociation of eNOS from caveolin 1 and its translocation to the cytosol in the active form, where it catalyzes the synthesis of NO. In fact, drugs interfering with sphingomyelinase activity blocked bFGF activation of eNOS, and an increase in ceramide content was detected after bFGF treatment. Moreover, in fibroblasts derived from patients with Niemann-Pick disease, in which the enzyme is genetically inactive, bFGF is unable to elicit eNOS activation. The NO produced after bFGF treatment, through the activation of guanylyl cyclase and protein kinase G, mediates a mitogen-activated protein kinase-independent cell proliferation. In conclusion, our data show that, in CHO-K1 cells, bFGF regulates the activity of eNOS through a novel intracellular pathway, involving the induction of ceramide synthesis and that the NO released participates in bFGF proliferative activity. PMID- 12527802 TI - Relating neuronal nicotinic acetylcholine receptor subtypes defined by subunit composition and channel function. AB - Neuronal nicotinic acetylcholine receptors (nAChRs) are widespread, diverse ion channels involved in synaptic signaling, addiction, and disease. Despite their importance, the relationship between native nAChR subunit composition and function remains poorly defined. Chick ciliary ganglion neurons express two major nAChR types: those recognized by alpha-bungarotoxin (alphaBgt), nearly all of which contain only alpha7 subunits (alpha7-nAChRs) and those insensitive to alphaBgt, which contain alpha3, alpha5, beta4, and, in some cases, beta2 subunits (alpha3*-nAChRs). We explored the relationship between nAChR composition and channel function using toxins recognizing alpha7 subunits (alphaBgt), and alpha3/beta4 (alpha-conotoxin-AuIB), or alpha3/beta2 (alpha-conotoxin-MII) subunit interfaces to perturb responses induced by nicotine, alpha7-, or alpha3 selective agonists (GTS-21 or epibatidine, respectively). Using these reagents, fast-decaying whole-cell current components were attributed solely to alpha7 nAChRs, and slow-decaying components mostly to alpha3*-nAChRs. In outside-out patches, nicotine activated brief 60- and 80-pS single nAChR channel events, and mixed-duration 25- and 40-pS nAChR events. Subsequently, 60- and 80-pS nAChR events and most brief 25- and 40-pS events were attributed to alpha7-nAChRs, and long 25- and 40-pS events to alpha3*-nAChRs. alpha3*-nAChRs lacking beta2 subunits seemed responsible for long 25 pS nAChR events, whereas those containing beta2 subunits mediated the long 40 pS nAChR events that dominate single-channel records. These results reveal greater functional heterogeneity for alpha7-nAChRs than previously expected and indicate that beta2 subunits contribute importantly to alpha3*-nAChR function. By linking structural to functional nAChR subtypes, the findings also illustrate a useful pharmacological strategy for selectively targeting nAChRs. PMID- 12527803 TI - Akt-dependent phosphorylation of serine 1179 and mitogen-activated protein kinase kinase/extracellular signal-regulated kinase 1/2 cooperatively mediate activation of the endothelial nitric-oxide synthase by hydrogen peroxide. AB - Hydrogen peroxide mediates vasodilation, but the mechanisms responsible for this process remain undefined. We examined the effect of H(2)O(2) on nitric oxide (NO*) production and the signaling events involved. NO* release from bovine aortic endothelial cells was detected with an NO*-specific microelectrode. The addition of H(2)O(2) caused a potent dose-dependent increase in NO* production. This was partially Ca(2+)-dependent because BAPTA/AM reduced NO* production at low (<50 microM) but not high (>100 microM) concentrations of H(2)O(2). Phosphatidylinositol (PI) 3-kinase inhibition [with wortmannin or 2-(4 morpholinyl)-8-phenyl-1(4H)-benzopyran-4-one hydrochloride], infection with a dominant-negative mutant of Akt, or mitogen-activated protein kinase kinase/extracellular signal-regulated kinase 1/2 (MEK/ERK1/2) inhibition (with PD98059 or U0126) partially attenuated, whereas inhibition of both PI 3-kinase and MEK1/2 abolished H(2)O(2)-dependent NO* production. ERK1/2 seemed necessary for NO* production early (<5 min) after H(2)O(2) addition, whereas PI 3 kinase/Akt was more important at later time points. Phosphorylation of endothelial nitric-oxide synthase (eNOS) at serine 1179 was observed >10 min after the addition of H(2)O(2), and this was prevented by wortmannin but not by PD98059. c-Src family tyrosine kinase(s) was found to be upstream of H(2)O(2) dependent Akt and eNOS serine 1179 phosphorylation and subsequent NO* production. In summary, H(2)O(2) causes endothelial NO* release mediated by cooperative effects between PI 3-kinase/Akt-dependent eNOS serine 1179 phosphorylation and activation of MEK/ERK1/2. This may represent an acute cellular adaptation to an increase in oxidant stress. PMID- 12527804 TI - Alternate stoichiometries of alpha4beta2 nicotinic acetylcholine receptors. AB - Two functional types of nicotinic acetylcholine receptors (nAChRs) are expressed when human embryonic kidney cells are permanently transfected with equal amounts of human alpha4 and beta2 subunit cDNAs. Most (82%) of these nAChRs exhibit an EC(50) of 74 +/- 6 microM for ACh, a much lower sensitivity than the remaining fraction (EC(50) of 0.7 +/- 0.4 microM) or than expected from expression of equal amounts of alpha4 and beta2 mRNAs in Xenopus laevis oocytes. We have found three conditions that can increase the number of nAChRs with high sensitivity to activation. These are: 1) transient transfection with additional beta2 subunits, 2) overnight incubation in nicotine, or 3) overnight culture at 29 degrees C. Using metabolic labeling with [(35)S]methionine to measure subunit stoichiometry, we found that the majority of nAChRs had a stoichiometry of (alpha4)(3)(beta2)(2). Overnight treatment with nicotine increased the number of nAChRs and increased the proportion of the (alpha4)(2)(beta2)(3) stoichiometry. Alternate alpha4beta2 nAChR stoichiometries with distinct functional properties raise the possibility for an interesting mode of synaptic regulation for nicotinic signaling in the mammalian brain. PMID- 12527805 TI - Pharmacological analysis of calcium responses mediated by the human A3 adenosine receptor in monocyte-derived dendritic cells and recombinant cells. AB - Extensive characterization of adenosine receptors expressed by human monocyte derived dendritic cells (MDDCs) was performed with quantitative polymerase chain reaction, radioligand binding, and calcium signaling. Transcript for the A3 adenosine receptor was elevated more than 100-fold in immature MDDCs compared with monocyte precursors. A3 receptor transcript was substantially diminished, and A2A receptor transcript increased, by lipopolysaccharide maturation of MDDCs. Saturation binding of N(6)-(3-[(125)I]iodo-4-aminobenzyl)-adenosine-5'-N methyluronamide ([(125)I]AB-MECA) to membranes from immature MDDCs yielded B(max) of 298 fmol/mg of protein and K(D) of 0.7 nM. Competition against [(125)I]AB-MECA binding confirmed the site to be the A3 receptor. Adenosine elicited pertussis toxin-sensitive calcium responses with EC(50) values ranging as low as 2 nM. The order of potency for related agonists was N(6)-(3-iodobenzyl)-adenosine-5'-N methylcarboxamide (IB-MECA) >/= I-AB-MECA > 2Cl-IB-MECA >/= adenosine > 2-[p-(2 carboxyethyl)phenylethylamino]-5'-N-ethylcarboxyamidoadenosine (CGS21680). The order of efficacy was adenosine >/= CGS21680 > IB-MECA >/= I-AB-MECA > 2Cl-IB MECA. Calcium responses to 2Cl-IB-MECA and CGS21680, and the lower range of adenosine concentrations, were completely blocked by 10 nM N-(2-methoxyphenyl)-N [2-(3-pyridyl)quinazolin-4-yl]urea (VUF5574) but not by 7-(2-phenylethyl)-5-amino 2-(2-furyl)-pyrazolo-[4,3-e]-1,2,4-triazolo[1,5-c]pyrimidine (SCH58261) or 8 cyclopentyl-1,3-dipropylxanthine. Pretreatment with 100 nM 2Cl-IB-MECA eliminated responses to CGS21680 but not to monocyte inhibitory protein-1alpha. For comparison, dose-response functions were obtained from double-recombinant human embryonic kidney 293 cells expressing the human A3 receptor and a chimeric Galphaq-i3 protein, which was required to establish A3-mediated calcium signaling. The pharmacological profile of calcium signaling elicited by adenosine related agonists in the double-recombinant cells was essentially identical to that obtained from immature MDDCs. Our results provide an extensive analysis of A3-mediated calcium signaling and unequivocally identify immature MDDCs as native expressers of the human A3 receptor. PMID- 12527806 TI - Characterization of the transport properties of human multidrug resistance protein 7 (MRP7, ABCC10). AB - Human multidrug resistance protein 7 (MRP7, ABCC10) is a recently described member of the C family of ATP binding cassette proteins (Cancer Lett 162:181-191, 2001). However, neither its biochemical activity nor physiological functions have been determined. Here we report the results of investigations of the in vitro transport properties of MRP7 using membrane vesicles prepared from human embryonic kidney 293 cells transfected with MRP7 expression vector. It is shown that expression of MRP7 is specifically associated with the MgATP-dependent transport of 17beta-estradiol-(17-beta-D-glucuronide) (E(2)17betaG). E(2)17betaG transport was saturable, with K(m) and V(max) values of 57.8 +/- 15 microM and 53.1 +/- 20 pmol/mg/min. By contrast, with E(2)17betaG, only modest enhancement of LTC(4) transport was observed and transport of several other established substrates of MRP family transporters was not detectable to any extent. In accord with the notion that MRP7 has a bipartite substrate binding pocket composed of sites for anionic and lipophilic moieties, transport of E(2)17betaG was susceptible to competitive inhibition by both amphiphiles, such as leukotriene C(4) (K(i(app)), 1.5 microM), glycolithocholate 3-sulfate (K(i(app)), 34.2 microM) and MK571 (K(i(app)), 28.5 microM), and lipophilic agents such as cyclosporine A (K(i(app)), 14.4 microM). Of the inhibitors tested, LTC(4) was the most potent, in agreement with the possibility that it is a substrate of the pump. The determination that MRP7 has the facility for mediating the transport of conjugates such as E(2)17betaG indicates that it is a lipophilic anion transporter involved in phase III (cellular extrusion) of detoxification. PMID- 12527807 TI - Nuclear factor-Y binding to the topoisomerase IIalpha promoter is inhibited by both the p53 tumor suppressor and anticancer drugs. AB - Expression of the human DNA topoisomerase IIalpha (topo IIalpha) gene is positively regulated by the binding of the nuclear factor Y (NF-Y) transcription factor to four of five inverted CCAAT boxes (ICBs) located in its promoter. We have demonstrated previously that expression of the p53 tumor suppressor inhibits human topo IIalpha promoter activity in murine (10)1 cells. In this report, we demonstrate that the inhibition of topo IIalpha gene expression by wild-type p53 correlates with the decreased binding of the transcription factor NF-Y to the first four ICBs of the topo IIalpha promoter. The expression of mutant p53 does not affect the binding of NF-Y. In NIH3T3 cells, we show that topo II-targeted drugs inhibit the binding of NF-Y to ICB sites in the topo IIalpha promoter. This effect is seen not only with drugs that result in DNA strand breaks but also with drugs that inhibit the catalytic activity of topo II, and even with the mitotic spindle inhibitor, vinblastine. Further experiments with p53-null (10)1 cells treated with these same drugs also demonstrate decreased NF-Y binding to the topo IIalpha ICBs. The data presented points to the existence of both p53-dependent and -independent mechanisms for regulating NF-Y binding to ICBs in the topo IIalpha promoter and thus the modulation of topo IIalpha gene expression. PMID- 12527808 TI - Anthracycline-induced suppression of GATA-4 transcription factor: implication in the regulation of cardiac myocyte apoptosis. AB - Anthracyclines are effective cancer chemotherapeutic agents but can induce serious cardiotoxicity. Understanding the mechanism of cardiac damage by these agents will help in development of better therapeutic strategies against cancer. The GATA-4 transcription factor is an important regulator of cardiac muscle cells. The present study demonstrates that anthracyclines can down-regulate GATA 4 activity. Treatment of HL-1 cardiac muscle cells or isolated adult rat ventricular myocytes with anthracyclines such as daunorubicin and doxorubicin decreased the level of GATA-4 DNA-binding activity. The mechanism of decreased GATA-4 activity acts at the level of the GATA-4 gene, because anthracyclines caused significantly decreased levels of GATA-4 protein and mRNA. The rate of decline in GATA-4 transcript levels in the presence of actinomycin D was unaltered by anthracyclines, indicating that these agents may affect directly GATA-4 gene transcription. To determine whether decreased GATA-4 levels are functionally related to cardiac muscle cell death that can be induced by anthracyclines, the ability of ectopic GATA factors to rescue anthracycline induced apoptosis was tested. Adenovirus-mediated expression of either GATA-4 or GATA-6 was sufficient to attenuate the incidence of apoptosis. Furthermore, suppression of GATA-4 DNA-binding activity by a dominant negative mutant of GATA 4 induced the apoptosis. These results suggest that the mechanism of anthracycline-induced cardiotoxicity may involve the down-regulation of GATA-4 and the induction of apoptosis. PMID- 12527809 TI - Norepinephrine release from the ischemic heart is greatly enhanced in mice lacking histamine H3 receptors. AB - We previously reported that histamine H(3) receptors (H(3)Rs) are present in cardiac sympathetic nerve endings (cSNE) of animals and humans, where they attenuate norepinephrine (NE) release in normal and hyperadrenergic states, such as myocardial ischemia. The recent creation of a transgenic line of mice lacking H(3)R provided us with the opportunity to assess the relevance of H(3)R in the ischemic heart. We isolated SNE from hearts of wild-type (H(3)R(+/+)) and knockout (H(3)R(-/-)) mice and found that basal NE release from H(3)R(-/-) cSNE was approximately 60% greater than that from H(3)R(+/+) cSNE. NE exocytosis evoked by K(+)-induced depolarization of cSNE from H(3)R(+/+) mice was attenuated by activation of either H(3)R or adenosine A(1) receptors (A(1)R). In contrast, NE release from cSNE of H(3)R(-/-) was unaffected by H(3)R agonists, but it was still attenuated by A(1)R activation. When isolated mouse hearts were subjected to ischemia for 20 min, NE overflow into the coronaries was 2-fold greater in the H(3)R(-/-) hearts than in those from H(3)R(+/+) mice. Furthermore, whereas stimulation of H(3)R or A(1)R reduced ischemic NE overflow from H(3)R(+/+) hearts by 50%, only A(1)R, but not H(3)R activation, reduced NE release in H(3)R(-/-). Our data demonstrate that NE release from cSNE can be modulated by various heteroinhibitory receptors (e.g., H(3)R and A(1)R) and that H(3)Rs are particularly important in modulating NE release in myocardial ischemia. Inasmuch as excessive NE release is clinically recognized as a major cause of arrhythmic cardiac dysfunction, our findings reveal a significant cardioprotective role of H(3)R on cSNE. PMID- 12527810 TI - Sporogen, S14-95, and S-curvularin, three inhibitors of human inducible nitric oxide synthase expression isolated from fungi. AB - The induction of human inducible nitric-oxide synthase (iNOS) expression depends (among other factors) on activation of the signal transducer and activator of transcription 1 (STAT1) pathway. Therefore, the STAT1 pathway may be an appropriate target for the development of inhibitors of iNOS expression. HeLa S3 cells transiently transfected with a gamma-activated site (GAS)/interferon stimulated response element-driven reporter gene construct were used as the primary screening system. Using this system, three novel inhibitors of interferon gamma-dependent gene expression, namely, sporogen, S14-95, and S-curvularin, were isolated from different Penicillium species. These three compounds also inhibited cytokine-induced, GAS-dependent reporter gene expression in stably transfected human A549/8-pGASLuc cells, confirming the data obtained with the above-mentioned screening system. Furthermore, in A549/8 cells, sporogen, S14-95, and S curvularin inhibited cytokine-induced activity of the human iNOS promoter [a 16 kilobase (kb) fragment in stably transfected A549/8-pNOS2(16)Luc cells], cytokine induced iNOS mRNA expression, and cytokine-induced nitric oxide (NO) production in a concentration-dependent manner. The proliferation of A549/8 cells, and the activity of the human eNOS promoter (a 3.5-kb fragment in stably transfected ECV pNOS III-Hu-3500-Luc cells), were only influenced marginally by the three compounds. Sporogen, S14-95, and S-curvularin also inhibited cytokine-induced activation of STAT1alpha in A549/8 cells. In conclusion, sporogen, S14-95, and S curvularin represent new transcriptionally based inhibitors of iNOS-dependent NO production, acting on the Janus tyrosine kinase-STAT pathway. These compounds may represent lead structures for the development of drugs inhibiting iNOS-dependent overproduction of NO in pathophysiological situations. PMID- 12527811 TI - Agonist but not antagonist ligands induce conformational change in the mouse aryl hydrocarbon receptor as detected by partial proteolysis. AB - The cytosolic transcription factor known as the aryl hydrocarbon receptor (AhR) undergoes transformation to a DNA-binding form by a series of processes initiated by binding of ligand. Subsequent steps include dissociation of several proteins that are complexed with the inactive receptor, nuclear translocation, and dimerization with Arnt. We have used limited proteolysis of the in vitro translated mouse AhR to determine whether this technique can detect conformational change(s) associated with AhR transformation and whether the effect of agonist and antagonist ligands can be distinguished by this assay. Limited digestion of [(35)S]AhR/AhR nuclear translocator (Arnt) by trypsin produced a peptide of approximately 40 kDa that was more resistant to proteolysis in the presence of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) than vehicle and was also Arnt-dependent. This trypsin-resistant peptide was also elicited in the presence of other agonist ligands, but not with antagonist ligands that do not form the DNA-binding AhR/Arnt complex. Immunoblot of trypsin-treated AhR/Arnt +/- TCDD indicated that the trypsin-resistant peptide did not include the N-terminal portion of the AhR against which the antibody was made. Truncated AhRs were also subjected to limited trypsinization. From AhR(1-399), a TCDD-dependent peptide of approximately 35 kDa was observed; from the constitutively active AhR(1-348), a band of approximately 30 kDa was produced from vehicle- and TCDD-treated protein. From these observations, we hypothesize that the trypsin-resistant peptide from full-length AhR spans approximately from amino acid 80 to 440. We conclude that agonist ligands initiate structural alteration in AhR that is Arnt-dependent and at least partially involves the ligand-binding/Per-Arnt-Sim domain. PMID- 12527812 TI - Critical role of c-Jun N-terminal protein kinase activation in troglitazone induced apoptosis of human HepG2 hepatoma cells. AB - The peroxisome proliferator-activated receptor agonist troglitazone (TRO) was used for treatment of non-insulin-dependent diabetes until its removal from the market because of its severe hepatotoxicity. However, the mechanism for its hepatotoxicity is still poorly understood. In this study, we investigated whether TRO caused cell death by altering signaling pathways associated with cell damage and survival in human hepatoma cells. Our data reveal that TRO caused time- and concentration-dependent apoptosis of HepG2 and Chang liver human hepatoma cells, as evidenced by DNA fragmentation and staining with Hoechst 33342. In contrast, 50 or 100 microM rosiglitazone, a structural analog of TRO, did not cause apoptosis in these hepatoma cells. TRO activated both c-Jun N-terminal protein kinase (JNK) and p38 kinase about 5-fold between 0.5 and 8 h before they returned to control levels at 16 h in HepG2 cells. In contrast, TRO failed to activate the extracellular signal-regulated kinase. Furthermore, TRO increased the levels of proapoptotic proteins, Bad, Bax, release of cytochrome c, and cleavage of Bid in a time-dependent manner. The antiapoptotic Bcl-2 protein level decreased in hepatoma cells treated with TRO. Pretreatment of hepatoma cells with a selective JNK inhibitor, anthra[1,9-cd]pyrazol-6(2H)-one (SP600125), significantly reduced the rate of TRO-induced cell death, whereas 4-(4-fluorophenyl)-2-(4 methylsulfinylphenyl)-5-(4-pyridyl)-1H-imidazole (SB203580), an inhibitor of p38 kinase, had little effect on apoptosis. Pretreatment with SP600125 also prevented JNK activation and c-Jun phosphorylation. In addition, rosiglitazone, which is not as toxic to hepatoma cells as TRO, did not stimulate JNK activity. Transfection of cDNA for the dominant-negative mutant JNK-KR (Lys-->Arg) or SEK1 KR (Lys-->Arg), an immediate upstream kinase of JNK, significantly reduced TRO induced JNK activation and cell death rate. Furthermore, SP600125 pretreatment effectively prevented the TRO-mediated changes in Bad, Bax, Bid cleavage, and cytochrome c release. These data strongly suggest that hepatotoxic TRO causes apoptosis by activating the JNK-dependent cell death pathway accompanied by increased Bid cleavage and elevation of proapoptotic proteins. PMID- 12527813 TI - Maurotoxin: a potent inhibitor of intermediate conductance Ca2+-activated potassium channels. AB - Maurotoxin, a 34-amino acid toxin from Scorpio maurus scorpion venom, was examined for its ability to inhibit cloned human SK (SK1, SK2, and SK3), IK1, and Slo1 calcium-activated potassium (K(Ca)) channels. Maurotoxin was found to produce a potent inhibition of Ca(2+)-activated (86)Rb efflux (IC(50), 1.4 nM) and inwardly rectifying potassium currents (IC(50), 1 nM) in CHO cells stably expressing IK1. In contrast, maurotoxin produced no inhibition of SK1, SK2, and SK3 small-conductance or Slo1 large-conductance K(Ca) channels at up to 1 microM in physiologically relevant ionic strength buffers. Maurotoxin did inhibit (86)Rb efflux (IC(50), 45 nM) through, and (125)I-apamin binding (K(i), 10 nM) to SK channels in low ionic strength buffers (i.e., 18 mM sodium, 250 mM sucrose), which is consistent with previous reports of inhibition of apamin binding to brain synaptosomes. Under similar low ionic strength conditions, the potency for maurotoxin inhibition of IK1 increased by approximately 100-fold (IC(50), 14 pM). In agreement with its ability to inhibit recombinant IK1 potassium channels, maurotoxin was found to potently inhibit the Gardos channel in human red blood cells and to inhibit the K(Ca) in activated human T lymphocytes without affecting the voltage-gated potassium current encoded by Kv1.3. Maurotoxin also did not inhibit Kv1.1 potassium channels but potently blocked Kv1.2 (IC(50), 0.1 nM). Mutation analysis indicates that similar amino acid residues contribute to the blocking activity of both IK1 and Kv1.2. The results from this study show that maurotoxin is a potent inhibitor of the IK1 subclass of K(Ca) potassium channels and may serve as a useful tool for further defining the physiological role of this channel subtype. PMID- 12527814 TI - Potentiation of alpha7-containing nicotinic acetylcholine receptors by select albumins. AB - Nicotinic receptors containing alpha7 subunits are ligand-gated ion channels widely distributed in the nervous system; they influence a diverse array of events because of their high relative calcium permeability. We show here that nicotine-induced whole-cell responses generated by such receptors can be dramatically potentiated in a rapidly reversible manner by some but not all albumins. The potentiation involves increases both in potency and efficacy with no obvious differences in rise and fall times of the response. The potentiation is not reduced by removing absorbed components; it is abolished by proteolysis, suggesting that the albumin protein backbone is essential. The fact that some albumins are ineffective indicates that minor differences in amino acid sequence may be critical. Experiments with open channel blockers indicate that the potentiation involves increased responses from active receptors rather than recruitment of receptors from a previously silent pool. Single channel recordings reveal that the potentiation correlates with increased single channel opening probability, reflected in increased frequency of channel opening and increased mean channel open time. The potentiation can be exploited to overcome blockade by noncompetitive inhibitors such as beta-amyloid peptide. The results raise the possibility that endogenous compounds use the site to modulate receptor function in vivo, and suggest that the receptors may represent useful targets for therapeutic intervention in cases where they have been implicated in neuropathologies such as Alzheimer's disease. PMID- 12527815 TI - Anandamide inhibits nuclear factor-kappaB activation through a cannabinoid receptor-independent pathway. AB - Anandamide (arachidonoylethanolamine, AEA), an endogenous agonist for both the cannabinoid CB(1) receptor and the vanilloid VR1 receptor, elicits neurobehavioral, anti-inflammatory, immunomodulatory, and proapoptotic effects. Because of the central role of nuclear factor-kappaB (NF-kappaB) in the inflammatory process and the immune response, we postulated that AEA might owe some of its effects to the suppression of NF-kappaB. This study shows that AEA inhibits tumor necrosis factor-alpha (TNFalpha)-induced NF-kappaB activation by direct inhibition of the IkappaB kinase (IKK)beta and, to a lesser extent, the IKKalpha subunits of kappaB inhibitor (IkappaB) kinase complex, and that IKKs inhibition by AEA correlates with inhibition of IkappaBalpha degradation, NF kappaB binding to DNA, and NF-kappaB-dependent transcription in TNFalpha stimulated cells. AEA also prevents NF-kappaB-dependent reporter gene expression induced by mitogen-activated protein kinase kinase kinase and NF-kappaB-inducing kinase. The NF-kappaB inhibitory activity of AEA was independent of CB(1) and CB(2) activation in TNFalpha-stimulated 5.1 and A549 cell lines, which do not express vanilloid receptor 1, and was not mediated by hydrolytic products formed through the activity of the enzyme fatty acid amide hydrolase. Chemical modification markedly affected AEA inhibitory activity on NF-kappaB, suggesting rather narrow structure-activity relationships and the specific interaction with a molecular target. Substitution of the alkyl moiety with less saturated fatty acids generally reduced or abolished activity. However, replacement of the ethanolamine "head" with a vanillyl group led to potent inhibition of TNFalpha induced NF-kappaB-dependent transcription. These findings provide new mechanistic insights into the anti-inflammatory and proapoptotic activities of AEA, and should foster the synthesis of improved analogs amenable to pharmaceutical development as anti-inflammatory agents. PMID- 12527816 TI - Apoptosis induced by (E)-5-(2-bromovinyl)-2'-deoxyuridine in varicella zoster virus thymidine kinase-expressing cells is driven by activation of c Jun/activator protein-1 and Fas ligand/caspase-8. AB - The molecular mode of cell killing by the antiviral drug (E)-5-(2-bromovinyl-2' deoxyuridine (BVDU) was studied in Chinese hamster ovary (CHO) cells stably transfected with the thymidine kinase gene (tk) of varicella zoster virus (CHO VZVtk). The colony-forming ability of the cells was reduced to <1% at a concentration of approximately 1 microM BVDU, whereas for nontransfected cells or cells transfected with tk gene of herpes simplex virus type 1 (CHO-HSVtk), a 1000 fold higher dose was required to achieve the same response. BVDU inhibited thymidylate synthase in CHO-VZVtk but not in CHO-HSVtk and control cells. On the other hand, the drug was incorporated into DNA of VZVtk- and HSVtk-expressing cells to nearly equal amounts. Because coexposure of CHO-VZVtk cells to exogenous thymidine protected them from BVDU-induced cell killing, the cells obviously die because of thymidine depletion. At highly cytotoxic BVDU doses (50 microM) and longer exposure times (24-48 h), VZVtk cells were blocked to some extent in S and G2/M phase and underwent apoptosis (48-72 h). Not only apoptosis but also necrosis was induced. The findings also show that the drug causes the induction of c-Jun and the activation of activator protein-1 resulting in increased level of Fas ligand (FasL) and caspase-8/-3 activation. Bid and poly(ADP-ribose) polymerase were cleaved by caspases. Expression of Bax increased, whereas Bcl 2/Bcl-x(L) remained unchanged. Transfection of dominant-negative Fas-associated death domain and inhibition of caspase-8 by N-benzyloxycarbonyl-IETD-fluoromethyl ketone strongly abrogated BVDU-induced apoptosis, indicating Fas/FasL to be crucially involved. Thus, BVDU-triggered apoptosis differs significantly from that induced by ganciclovir, which induces in the same cellular background the mitochondrial damage pathway. PMID- 12527817 TI - A novel family of hydroxamate-based acylating inhibitors of cyclooxygenase. AB - Aspirin irreversibly inhibits cyclooxygenase (COX) by acetylating a serine residue in the active site. We synthesized a series of novel acylating agents based on our previously reported acetylating compound, O-acetylsalicylhydroxamic acid. One of these, triacetylsalicylhydroxamic acid (TriAcSHA) was more effective than aspirin and O-acetylsalicylhydroxamic acid in inactivating both COX-1 and COX-2. Preincubation of COX-1 with inhibitor for 5 min yielded IC(50) values of 18 microM for TriAcSHA and 60 microM for acetylsalicylic acid. Inhibition was time-dependent, with complete inhibition within 10 min at a concentration of 50 microM. As with aspirin, mutation of the serine 530 of COX-1 to alanine abolished the activity of the TriAcSHA. Mutation of the alanine 119 to a glutamine markedly reduced the sensitivity to TriAcSHA, suggesting that this residue was necessary for the interaction with the enzyme. TriAcSHA was also more effective than aspirin as an inhibitor of platelet aggregation induced by arachidonic acid. The diacetylated phenylhydroxamates N-methyl-O,O-diacetylsalicylhydroxamic acid, N,O diacetylbenzohydroxamic acid, and 2-methyl-O,N-diacetylbenzohydroxamic acid showed reduced or absent activity against COX-1. In addition, we synthesized a series of triacylsalicylhydroxamic acids with progressively longer acyl groups (three to six carbons). All of the compounds inhibited COX-1 and demonstrated progressively greater COX-1 selectivity with increasing number of carbons. Hence, salicylhydroxamic acid provides a versatile backbone for the generation of a family of acylating inhibitors of cyclooxygenase. PMID- 12527818 TI - In vivo evidence for dopamine-mediated internalization of D2-receptors after amphetamine: differential findings with [3H]raclopride versus [3H]spiperone. AB - Competition with endogenous dopamine (DA) is usually invoked to explain changes in [(11)C]raclopride binding observed after amphetamine administration in animals and humans. This account has recently been questioned because a number of inconsistencies have been reported that contradict it. In the present study, we investigated whether the decrease in [(3)H]raclopride binding observed in the rat striatum after an amphetamine challenge reflects true competition with endogenous DA or agonist-mediated internalization of D(2)-receptors. We found that the amphetamine-induced decrease in [(3)H]raclopride binding is caused by a decrease in D(2)-receptor density (B(max)) with no change in affinity (K(d)). In contrast, in the same tissue, neither the B(max) nor the K(d) were affected when measured with [(3)H]spiperone. Challenge with amphetamine not only decreased the number of D(2)-receptors but also eliminated the proportion (22%) of receptors usually in the high-affinity state. The addition of Gpp(NH)p had no effect on B(max), suggesting that these receptors were not just noncompetitively bound with dopamine at the cell-surface. Subcellular fractionation studies showed that amphetamine treatment led to a decrease in radioligand binding in the cell surface fraction for both [(3)H]raclopride and [(3)H]spiperone; however, in the case of [(3)H]spiperone, this was accompanied by a compensatory increase in binding in the intracellular compartment, whereas no increase was seen with [(3)H]raclopride. These data suggest that amphetamine releases dopamine, which binds to the high-affinity state of the D(2)-receptor, leading to its sequestration in some intracellular compartment; in this compartment, sequestered receptors are inaccessible to [(3)H]raclopride binding but can still be bound by [(3)H]spiperone. PMID- 12527819 TI - DARPP chocolate: a caffeinated morsel of striatal signaling. AB - The psychomotor stimulant effects of caffeine, the most widely consumed psychoactive substance, are mediated through its antagonism of extracellular adenosine receptors in the basal ganglia. In the absence of caffeine, adenosine stimulates inhibitory striatopallidal neurons that suppress motor activity by binding to A2A receptors, thereby activating a cyclic adenosine 3',5' monophosphate (cAMP) and protein kinase A signaling pathway. Bastia and Schwarzschild discuss recent research implicating DARRP-32 (dopamine- and cAMP regulated phosphoprotein of 32 kilodaltons) as an attractive mediator of the sustained psychomotor stimulant effect seen with low doses of caffeine. They highlight the role of postsynaptic A2A receptor blockade, but leave open the possibility that antagonism of presynaptic or postsynaptic A1 receptors also contributes to DARPP-32-dependent psychomotor stimulation by caffeine. PMID- 12527820 TI - Shedding light on cell signaling: interpretation of FRET biosensors. AB - This Perspective compares recently developed approaches for studying live-cell signaling dynamics. It is now possible not only to study the changing localizations of proteins within living cells but to correlate these dynamics with quantitation of protein activities, including ligand interactions and phosphorylation. Initial applications of an increasingly varied toolchest of techniques have revealed strengths and weaknesses in each approach, clarifying which tool is best for which job. PMID- 12527821 TI - [N acetyl-aspartyl glutamic acid (NAAGA) inhibits the adhesion of leukocytes to activated endothelial cells and down-modulates the cytokine-induced expression of adhesion molecules]. AB - BACKGROUND: Cell adhesion plays a pivotal role in most ocular surface inflammatory diseases. Adhesion molecules mediate cell-to-cell and cell-to-matrix adhesion. Their expression is up-regulated by pro-inflammatory stimuli such as cytokines, histamine or complement-derived anaphylatoxins. The dipeptide N acetyl aspartyl glutamic acid (NAAGA) is used as unpreserved topical eyedrops in the treatment of allergic conjunctivitis. NAAGA is known to inhibit leukotriene synthesis, histamine release by mast cells, and complement-derived anaphylatoxin production. PURPOSE: To investigate the potential capability of NAAGA to interfere with leukocyte adhesion to endothelial cells and modulate cytokine induced expression of adhesion molecules. METHODS: Human blood-derived leukocytes were co-cultured with human umbilical vein endothelial cells (HUVECs) in the absence or the presence of 1000 UI/mL human recombinant TNFalpha, 10(-4) M histamine di-hydrochloride or 5x10(-6) M human recombinant C5a, and in the absence or presence of NAAGA (final concentration 2.45%). Adhesion of leukocytes to HUVECs was calculated by subtracting the number of nonadherent leukocytes from the total number of leukocytes. Expression of adhesion molecules was assessed by flow cytometry using anti-CD11b, anti-CD49d, anti-ICAM-1 (CD54), anti-ICAM-2 (CD102), anti-VCAM-1 (CD106) and anti-ELAM-1 (CD62E) monoclonal antibodies. RESULTS: NAAGA was found to totally inhibit adhesion of unstimulated leukocytes, or leukocytes activated with C5a, TNFalpha, or histamine, to TNFalpha-stimulated HUVECs (P=0.0001). Adhesion of leukocytes to unstimulated HUVECs was not modified by NAAGA. Similar results were obtained with endothelial cells stimulated by histamine or C5a. Taken together, these data indicate that NAAGA totally abrogates cell adhesion under inflammatory conditions, without interfering with the physiological adhesion of leukocytes to normal endothelium. At the molecular level, NAAGA inhibited histamine-induced expression of CD11b (P=0.0004) and CD49d (P=0.0045) on granulocytes. On TNFalpha-activated HUVECs, NAAGA induced a significant decrease in the VCAM-1 expression level (P<0.0001) and totally reversed TNFalpha-induced overexpression of ICAM-1 (P=0.0069), ICAM-2 and ELAM-1 (P<0.0001), without interfering with baseline expression of these molecules. CONCLUSION: These results show that the antiallergic compound NAAGA directly inhibits leukocyte adhesion to endothelial cells induced by pro-inflammatory stimuli, and abrogates TNFalpha-induced expression of adhesion molecules on granulocytes and endothelial cells. This capacity to block overexpression of selectins and integrins induced by pro-inflammatory stimuli confers to NAAGA a potential as an anti-inflammatory drug, since interfering with adhesion molecule expression is probably one of the most efficient ways to curb leukocyte recruitment to inflammatory sites. PMID- 12527822 TI - [Episcleral Miragel buckling elements removal. A clinical case-series]. AB - INTRODUCTION: Due to a long-term alteration in their chemical composition, resulting in excessive swelling, Miragel scleral buckles are now commonly removed. The aim of this study was to determine the characteristics and the outcome of patients needing Miragel buckling removal. METHODS: Retrospective analysis between 1997 and 1999 of the charts of patients successfully treated for retinal detachment and in whom a Miragelepiscleral buckle was removed after surgery with a minimal follow-up of 6 months after removal. RESULTS: The study group consisted of 24 eyes of 13 women and 11 men. Their mean age at removal was 47.1+/-18.4 years. The removed buckle was encircling in nine cases (37.5%), circumferential (<180 degrees ) in 11 cases (45.8%) and radial in four cases (16.7%). An exposed buckle was observed in two cases (8.3%). Reasons for removal were excessive swelling of the buckle inducing ocular motility disturbance in 23 cases and infection of an extruded buckle in only one case. The mean time between implantation and removal was 91.9+/-28.7 months (range 20-120). One case of scleral perforation during removal and one case of retinal redetachment were observed. Three cases of persistent ocular motility disturbance after removal were also noted. CONCLUSION: Late swelling of the buckle was the main reason for removal. Patients undergoing Miragel buckling removal should be informed of scleral perforation during removal, retinal redetachment and persistence of ocular motility disturbance as possible complications. PMID- 12527823 TI - [Clinical management and prognosis in Acanthamoeba keratitis: a retrospective study of 25 cases]. AB - PURPOSE: To assess risk factors, clinical characteristics, diagnosis and outcome in patients with Acanthamoeba keratitis. METHODS: We reviewed 23 patients (25 eyes) treated for Acanthamoeba keratitis in the XV-XX Hospital between November 1994 and October 2000. Acanthamoeba were found either in corneal scrapings and/or contact lenses and/or storage solutions. RESULTS: A predisposing factor was found in 22/23 patients: most of them (20/22) were contact lens (CL) wearers, two patients had a recent history of corneal trauma, and 15/23 patients had been treated for presumed herpetic keratitis before examination in our hospital. The average delay between first symptoms and diagnosis was 2 months. All patients were treated with two or three topical antiparasitic eye drops for 2 to 6 months. Five patients received systemic treatment (ketoconazole or itraconazole). Nine eyes received surgical treatment: penetrating keratoplasty in eight cases, conjunctival flap in one case, enucleation in one case. Visual outcome was poor in ten eyes (less than 20/60) because of delayed diagnosis. CONCLUSION: Acanthamoeba keratitis is a serious vision-threatening disease. Early diagnosis and treatment are essential for improving visual outcome. Methods such as confocal microscopy and Acanthamoeba-PCR, allowing earlier diagnosis and treatment, will improve the management of Acanthamoeba keratitis. PMID- 12527824 TI - [Etiologic causes of ptosis about a serie of 484 cases. To a new classification?]. AB - We have analyzed the etiological causes of ptosis in a retrospective study of 484 cases undergoing operation. This study has confirmed how difficult it is to classify ptosis despite the different existing classifications. Some authors divide ptosis into two groups: congenital and acquired. These classifications seem to have been abandoned now for classifications based on the mechanism that instigates ptosis rather than the moment when the deficit developed. We chose to classify ptosis into five subgroups: myogenic (42% of the cases studied), aponeurotic (35.3%), neurogenic (6.8%), mixed (15.9%), and pseudoptosis (enophthalmos, eyelid tumor, hypotropia, etc.), the latter of which were removed from this series. This classification is based on clinical and surgical criteria. It has the advantages of unity, simplicity, and practicality in terms of establishing a treatment plan for a given ptosis patient. Indeed, each subgroup requires a particular clinical examination and a more stereotyped surgical treatment. PMID- 12527825 TI - [Helicobacter pylori in central serous chorioretinopathy and diffuse retinal epitheliopathy. Results of the first prospective pilot study]. AB - PURPOSE: Helicobacter pylori has been implicated in focal occlusive arterial diseases in young people. Central serous chorioretinopathy (CSC) and diffuse retinal epitheliopathy (DRE) being suspected vascular occlusive diseases of choriocapillaris, the purpose of this study was to determine the prevalence of H. pylori infection in patients with long-lasting (>6 months) CSC and/or DRE. METHODS: Sixteen consecutive patients living in southeastern France were included in this prospective pilot study. H. pylori infection was assessed by the (13)C urea breath test, serology, as well as histology of gastric biopsy specimens in some cases. RESULTS: Evidence for H. pylori infection was detected in nine out of the 16 patients with active long-lasting CSC/DRE (56.3%). H. pylori-infected patients more frequently presented gastric pain (four of nine H. pylori-positive patients: 44%) than H. pylori-negative patients (one out of seven: 14%). Men were more frequently H. pylori-positive (seven men out of ten: 70%) than women (two women out of a total of six: 33%). DISCUSSION: When the prevalence of H. pylori infection (56.3%) was compared to the prevalence in a historical control population from southeastern France (27.5%), it was found to be significantly higher (P<0.05). CONCLUSION: These results may indicate a possible association between H. pylori infection and CSC/DRE manifestations. This new etiopathogenic hypothesis deserves to be confirmed in a national or international multicenter study because it could lead to a new therapeutic approach in CSC/DRE, i.e., Helicobacter pylori eradication. PMID- 12527826 TI - [Study of internal limiting membrane peeling in stage-3 and -4 idiopathic macular hole surgery]. AB - PURPOSE: To evaluate the advantages of peeling the internal limiting membrane (ILM) in macular hole surgery. PATIENTS AND METHODS: Retrospective analysis of the results of macular hole surgery, comparing two surgical protocols performed by the same surgeon. Both protocols included a standard pars plana vitrectomy with intravitreous nonexpansile gas and adjunction of autologous plasma over the macular hole. In addition, in the second group, the ILM of the eyes was systematically peeled after its coloring by indocyanine green (infracyanine) in the latter patients. RESULTS: We studied 39 eyes of 36 patients with stage-3 or 4 macular hole surgery. The duration of symptoms before surgery was on average 9 months. Twenty-one eyes (53.8%) underwent ILM peeling. Macular hole closure after one procedure was significantly more frequent postoperatively in the group of eyes with ILM peeling than in the other group (90% and 50%, respectively, p<0.01); after two procedures, macular hole closure reached 61% in the second group. The macular hole seemed to have disappeared on angiographic examination and on OCT in 90% of eyes with anatomical success in the group with ILM peeling and in only 22% of eyes with anatomical success in the group without. Visual acuity improved by two lines or more in 62% of eyes with ILM peeling and in 44% of eyes without. The difference in improvement in the macular threshold was statistically significant (p<0.001). CONCLUSION: This study would suggest that peeling of the ILM is an important adjuvant for successful closure of macular holes. Its possible mechanism of action is discussed. Further studies are needed to confirm these data by controlled randomized trial. PMID- 12527827 TI - [Bilateral diffuse uveal melanocytic proliferation associated with systemic carcinoma: two case reports]. AB - Diffuse uveal melanocytic proliferation is a rare paraneoplastic syndrome resulting in rapid bilateral visual loss in patients with systemic carcinoma, caused by proliferation of benign melanocytes within the choroid and the ciliary body. More often visual impairment is due to retinal detachment and cataract. The authors report two cases of presumed diffuse uveal melanocytic proliferation. The first patient was a 74-year-old man with a history of colic carcinoma and hemangioendothelioma of the liver who presented with bilateral multiple nevi of the choroid and extrascleral melanic nodule. The second patient was a 59-year-old woman who presented bilateral multiple nevi of the choroid and a history of carcinoma of the thyroid treated by thyroidectomy 2 years before. There was no evidence of systemic melanoma in either patient. Our two patients showed slow progression with no visual impairment and a longer survival than those described in the literature. PMID- 12527828 TI - [Malignant fibrous histiocytoma of the eyelid]. AB - Fibrous histiocytomas, mostly benign, have often been reported in the orbit, but only four lesions have been described on the eyelids, with only one malignant. The authors present a case of malignant fibrous histiocytoma of the eyelid in a 76-year-old woman with a difficult histological diagnosis. Characteristics of this pathology are explained from data in the literature. PMID- 12527829 TI - [Bilateral choroidal metastasis revealing disseminated cancer: a case report]. AB - We report the case of a 46-year-old woman, with no prior medical history, who developed a metamorphopsia of the right eye, followed by visual impairment. Ophthalmological examination revealed bilateral choroidal metastasis. Further investigations led to the conclusion of multifocal metastasis of bronchial adenocarcinoma. PMID- 12527830 TI - [Intraocular pressure elevation after subtenon injection of triamcinolone acetonide during uveitis]. AB - INTRODUCTION: New therapeutic concepts in the management of ocular inflammation have led to the development of periocular and intravitreal injections. Such treatment modalities can induce intraocular pressure elevation. PATIENTS AND METHODS: Periocular injections have been given to patients suffering from strictly unilateral or bilateral but asymmetrical and noninfectious posterior uveitis. A history of corticosteroid-induced glaucoma was a contraindication to such treatment. A retrospective review of cases who were given subtenon triamcinolone acetonide injection between May and October 2001 was undertaken to evaluate the efficacy of the treatment and the risk of intraocular pressure elevation. Ocular pressure was measured before and after the injection and the efficacy of the treatment was evaluated by measurements of visual acuity and fluorescein angiography. RESULTS: One or several injections were given to 61 patients. Intraocular pressure rose in 13 patients (21.3%). Medical treatment was unsuccessful in three cases and surgical excision of periocular corticosteroid deposit was required. Therefore, intraocular pressure was controlled with no other medication. Treatment was considered effective in 32 patients (52.45%): improvement of visual acuity (more than two lines) or control of inflammation on fluorescein angiography. DISCUSSION: and conclusions: Periocular subtenon injection of triamcinolone acetonide in posterior noninfectious uveitis is a safe procedure. Intraocular pressure elevation is not frequent and can be controlled through medical treatment or surgical excision of a residual deposit, in which pharmacologically active triamcinolone can be present several months after the injection. PMID- 12527831 TI - [Value and practice of in-home "low-vision" rehabilitation in Rhone-Alp region of France]. AB - INTRODUCTION: To demonstrate the interest of patient's home orthoptist deplacement to realize an in-home low-vision rehabilitation and encourage orthoptists to integrate this task in their usual work methods. PATIENTS AND METHODS: Eighty-five consecutive patients coming from a private ophthalmic rehabilitation Center and needing low-vision rehabilitation, were studied. The exercises used at patient's home were usually the same as those used in the rehabilitation Center. The choice in the organization of rehabilitation was always function of patient general health, people vicinity, and social context. RESULTS: Among the 85 patients, 60 (70.6%) were rehabilitated only in the ophthalmic Center, 11 (12.9%) could have a low-vision rehabilitation only thanks to the organization of the home's patient rehabilitation and 14 (16.5%) were both rehabilitated at the ophthalmic Center and, in the patient's home, some training was necessary too. The reasons for in-home low-vision rehabilitation were mainly represented by isolation problems, mobilisation physical capacities, but also, home installation conditions making difficult to realize the orthoptist recommendations. DISCUSSION: Besides some disadvantages: time-consuming, badly paid, decrease of efficiency in familiar environment, advantages are easily objected: ergonomy, training on optic systems, and psychological comfort. CONCLUSION: In-home low-vision rehabilitation is represented by the necessity of correctly taking care of each patient. It is an answer to a social demand and must be an integral part of low-vision orthoptist work. PMID- 12527832 TI - [Limbal stem cell deficiency associated with KID syndrome, about a case]. AB - We report the case of a 30-year-old woman with KID (keratitis ichthyosis deafness) syndrome consulting for massive corneal neovascularization. She had already undergone two unsuccessful penetrating keratoplasties in her left eye. Visual acuity was limited to no more than light perception because of the opacity and neovascularization of the graft. In the light of this case of limbal cell deficiency, we discuss the ophthalmologic manifestations of KID syndrome as well as its clinical characteristics, physiopathology, and mode of transmission. PMID- 12527834 TI - [Morphological imagery of the occipital lobe]. AB - After reviewing the anatomical and functional aspects of the occipital lobe, the authors detail its gyral and sulcal anatomy using MRI in the three planes, with particular attention given to the localization of the calcarine fissure. PMID- 12527835 TI - [Vision disorders of central origin]. AB - Cortical blindness (or more generally, cerebral blindness) consists of bilateral visual loss with normal ocular examination and normal pupillary function, related to bilateral occipital lesions. Its most frequent cause in adults is hypoperfusion in the vertebral-basilar arteries, although other curable causes are described. Other syndromes, related to lesions of associative cortical areas or occipital-fugal connections may cause poorly defined visual complaints. Their recognition by the ophthalmologist is mandatory, as they may concern a potentially curable underlying disease. PMID- 12527836 TI - [Methylene blue and tear duct replacement]. PMID- 12527837 TI - [Actinic keratosis seborrheic wart and nevus: three distinct cutaneous eyelid skin lesions in a single patient]. AB - A 67-year-old woman had three different tumors of the eyelids. The most prominent one was a nevus of the free margin of the inferior left eyelid. The nevus was followed for 18 months without change while two epidermal papillomatous tumors grew and were surgically removed. Histopathology disclosed that one tumor was a seborrheic keratosis and the other was a pre-epitheliomatous keratosis. Histologic evaluation of the margins showed that surgical removal was complete. A regular examination of the scars was recommended to the patient. PMID- 12527838 TI - [Should open-angle glaucoma be treated?]. PMID- 12527839 TI - [Scintigraphy by l'Octreoscan in the management of head and neck paragangliomas]. AB - OBJECTIVE: To assess the usefulness of somatostatin receptor scintigraphy [Octreoscan] in a series of 18 patients referred for a suspicion of paraganglioma of the head and neck between July 2001 and February 2002. PATIENTS AND METHODS: Sixteen patients had one or several paragangliomas of the head and neck diagnosed on conclusive conventional imaging including CT and MR scan. In two patients, radiological data were not conclusive. Planar images were obtained 4 and 24 hr after the iv injection of 148-185 MBq [Octreoscan]. RESULTS: Twenty-two hot spot lesions were detected. Twenty of these lesions corresponded to the twenty known paragangliomas. The volume of the smallest tumor was 0.2 cm(3). In one patient, intense thyroid nodule uptake led to the surgical diagnosis of oncocytoma. In two lesions, where conventional imaging was not conclusive, arteriography showed a typical aspect of meningioma; one patient was operated on and histology confirmed this diagnosis. No evidence of abnormal uptake was seen in site previously operated on (3 patients). CONCLUSION: Octreotide scintigraphy is a very sensitive method for detection of paraganglioma of head and neck. It provides information on potential tumor sites in the whole body after one single injection. It could be used as a screening test in patients at risk (familial or known paraganglioma) in order to detect paraganglioma at an early stage and, thus to reduce the surgical morbidity, as well as in the follow-up after surgery to detect recurrences. PMID- 12527840 TI - [Analysis of eosinophilia and ECP levels in blood and nasal secretions of 119 nasal polyposis patients]. AB - OBJECTIVE: The aim of this prospective study was to determine eosinophilia and ECP (Eosinophilic Cationic Protein) levels in blood and in nasal secretions of patients with nasal polyposis (NP). PATIENTS: 119 patients with NP were prospectively studied. The control group included 25 patients. METHODS: They included: questionnaire about asthma and intolerance to aspirin; nasal endosopic grading; nasal symptoms scoring; allergy testing; measurements of serum and nasal values of eosinophilia expressed as a percentage; measurements of serum and nasal values of ECP expressed in ng/ml; the Spearman correlation test, the T series and Khideux tests were used in the statistical analysis. RESULTS: In NP group, 66 were asthmatic (As) and 53 non asthmatics (NAs), 40 were atopic (ATo), and 79 were non atopic (NAto). Values of nasal eosinophilia and nasal ECP in NP patients were significantly higher (respectively 40.7+/-35.6% and 22.8+/-48.0 ng/ml) than the control group (respectively 3.0+/-14% and 2.1+/-3.6 ng/ml). Nasal and serum eosinophilia values in asthmatic patients (As) (respectively 47+/-36% and 7.6+/ 6.3%) were significantly higher (p<0.05) than in non asthmatic patients (NAs) (respectively 32.6+/-33.8% and 4.6+/-3.1%). No difference was found in nasal and serum ECP values between asthmatic and non asthmatic patients. Nasal eosinophilia values were significantly correlated (p=0.02) with nasal scoring in NP patients. No correlation was found between nasal ECP and serum ECP values and clinical scoring. CONCLUSION: Nasal eosinophilia and nasal ECP values are significantly increased in NP. Nasal eosinophilia seems to be a relevant biological marker of clinical severity (association with asthma and a high clinical score) in NP patients. By contrast, nasal ECP cannot be consider as a marker of clinical severity. PMID- 12527841 TI - [Early otorhinolaryngological manifestations of Wegener's granulomatosis. Analysis of 21 patients]. AB - Wegener's Granulomatosis (WG) is a necrotizing granulomatous vasculitis that has a strong affinity for the upper respiratory tract. OBJECTIVE: To retrospectively study the clinical features of otorhinolaryngological manifestations from 21 WG patients. PATIENTS: Eleven men and 10 women were studied with respectively a mean age of 62.7 (23-79) and 63.9 (53-73). METHODS: Otorhinolaryngological manifestations were recorded before and during the course of WG. RESULTS: Upper respiratory tract involvement occurred in 81% of cases (17/21 patients) and was isolated in 42.8% of cases (9/21 patients). Nasosinusal manifestations occurred in 71.4% of cases (15/21) before and at the time of GW diagnosis. They included bilateral sinusitis (7 cases), nasal crusting (6 cases), purulent rhinorrhea (5 cases), epistaxis (4 cases), nasal ulcers (2 cases), nasal congestion with obstruction (3 cases) and 4 cases of nose deformity (saddle nose or oedema). Otologic manifestations occurred in 28.5% of cases and were never isolated. They included otitis media (3 cases), sudden hearing loss (3 cases), tinnitus (1 case), facial palsy (1 case) and 2 cases of chondritis. Pharyngolaryngotracheal manifestations occurred in 33.3% of cases (7/21). Diagnosis of GW was based on positive ANCA test (95.2% of cases), presence of biologic inflammatory parameters (76% of cases) and histological features. 29 biopsies from nasosinusal lesions on 17 patients were made. 44.8% of the biopsies were contributive with at least one histologic feature of the combination including vasculitis, necrosis and granulomatous inflammation. The best contributive site of biopsy was the paranasal sinus. CONCLUSION: We report that otorhinolaryngological manifestations occurred in 81% of cases before and at the time of GW diagnosis. These findings indicate that otorhinolaryngologists have a central role to play in early diagnosis of the disease. PMID- 12527842 TI - [Middle ear adenoma: report of two cases and review of the literature]. AB - INTRODUCTION: Middle ear adenoma is an uncommon tumor. We report two cases and review the relevant literature concerning this rare entity. MATERIAL AND METHODS: Two patients with a middle ear adenoma were operated on in our department. Pre- and postoperative data concerning clinical examination, audiometry, and radiology were obtained. Information concerning intraoperative observations and the pathological examination were also gathered. RESULTS AND DISCUSSION: The diagnosis of middle ear adenomas is based on clinical, radiological, and pathological confrontation. The microscopic examination should be combined with histochemical and immuno-histochemical methods in order to evidence the glandular and neuroendocrin components of these lesions. CONCLUSION: Today, classified as a benign epithelial tumor, the middle ear adenoma is associated with an excellent prognosis provided total excision. PMID- 12527843 TI - [Attical fixation of the malleus head associated with obstruction of the round window niche by a high jugular bulb]. AB - OBJECTIVE: To report the case of a 52 year-old-woman presenting with a left mixed hearing loss. Surgical exploration revealed an attical fixation of the malleus head associated with obliteration of the round window by a high jugular bulb. METHOD: Surgical exploration revealed this rare association. Attical fixation of the malleus head was treated by a partial prosthesis. The mobility of the chain was normal at the end of the surgical exploration but it was not possible to search for the round window sign because the niche of the round window was obliterated by an high Jugular bulb. RESULTS: Audiogram was unchanged even if CT scan confirm the good position of the partial prosthesis. CONCLUSION: Surgical treatment of an attical fixation of the malleus head do not seem to be necessary if it is associated with a simultaneous obliteration of the round window. Indeed, obstruction of the round window by a high Jugular bulb can caused a conductive hearing loss. PMID- 12527844 TI - [Intrapetrous aneurysms of internal carotid artery: therapeutic considerations: two cases]. AB - Two cases of internal carotid artery aneurysm arising within the petrous bone are described, bringing up to 56 the number of such cases reported to date in the medical literature scanned through Medline. The first case presented as a moderately bleeding tumor of the petrous apex, while the second was revealed by a massive otorrhage. In both cases a pulsatile nasopharyngeal mass was associated with otorrhage. The treatment required in both cases the aneurysm to be embolized in extreme emergency. One patient suffered transient hemiplegia and aphasia that completely recovered, while the second eventually died, although the carotid occlusion had been well tolerated in the first two weeks that followed this procedure. From their own experience and a literature review, we can emphasize the need for arterial extra-intracranial bypass before an aneurysm embolization be proposed, since the neurological issue following internal carotid occlusion cannot be predicted. PMID- 12527845 TI - [Middle ear metastasis. One case report]. AB - Metastatic carcinoma involving the middle ear space is a rare entity that is little described in the literature. The number of cases seems to be increasing. It is usually recognized that palliative treatment as the most reasonable treatment for this tumors. CASE REPORT: A 59-year-old female patient presented with a history of a metastatic carcinoma involving the middle ear space and mastoid. This metastase occurred one year late in the course of a bladder cancer. Presentation of the lesion included otorrhea and facial palsy complicated by headache and bilateral visual loss in relation with lateral sinus thrombophlebitis and intracranial hypertension. Diffuses metastases in the skeletal system were demonstrated by pretreatment investigations. Treatment was only palliative chemotherapy. Outcome was fatal two months after onset of symptoms. DISCUSSION: This case illustrates the clinical and pathological features of this tumor and the pitfalls of diagnosis and treatment. We will discuss the current accepted management and the difference after diagnosis modalities for this type of metastatic tumor. PMID- 12527846 TI - [Venous malformations of the parotid in adults. Report of two cases and review of the literature]. AB - Parotid vascular malformation in the adult is a very rare and benign tumor; only 47 cases were described in the world literature. This vascular malformation, mostly of venous origin is, on the opposite of the clinical, histological and evolution features of parotid hemangiomas in children, which are more frequent. Some clinical and radiological elements are pathognomonic allowing preoperative diagnosis. We present 2 cases of intraparotid venous malformation in adults, and based on an exhaustive study of the world literature discuss frequency, diagnosis and treatment of this disease. PMID- 12527847 TI - [Ewing's sarcoma of the head and neck: a case report]. AB - We report a case of Ewing's sarcoma located in the ethmoid sinus. The patient complained of facial pain, anosmia and visual defect. Diagnosis was established at microscopic examination with histoimmunochemistry and molecular biology. Treatment combined chemotherapy and surgical resection with skull base reconstruction and post-operative ratio and chemotherapy. PMID- 12527848 TI - [Relapsing polychondritis with laryngotracheal extension: therapeutic approach in one patient]. AB - OBJECTIVE: We report the case of a 32-years-old woman suffering from relapsing polychondritis in a laryngeal localization leading to dyspnea due to stenosis or collapse. METHODS: Incomplete efficiency of medical treatment required endoscopic insertion of a laryngotracheal silicone prosthesis (Trachobronxane ST, Novatech) which avoided tracheotomy and allowed better upper airway patency. Clinical and endoscopic follow-up was continued for several months. RESULTS: The prosthesis restored laryngotracheal patency and allowed moderate physical activity. The follow-up endoscopic explorations showed excellent prosthesis tolerance eight months after implantation. No deglutition problems, due to the supraglottic position of the upper extremity of the prosthesis, was noted. CONCLUSION: Silicone prostheses provide an interesting alternative for the management of laryngotracheal collapse or stenosis. Withdrawal can be proposed if the disease processes is controlled. PMID- 12527850 TI - [Circadian rhythm of cortisol and its responsiveness to ACTH during Ramadan]. AB - Changes in the nyctemeral pattern observed during Ramadan include diurnal fasting from sunrise to sunset and delayed and shortened periods of sleep. The purpose of the present study was to investigate the possible effect of these changes on the circadian rhythm of cortisol and its response to exogenous ACTH. SUBJECTS AND METHODS: Eleven healthy male volunteers aged 20 to 35 years were studied for two weeks prior to Ramadan. Two short ACTH stimulation tests (250 microg ACTH 1-24 intravenously) at 8 a.m. and 8 p.m. An average 17 days after the beginning of the month of Ramadan, basal cortisol was measured at 8 a.m. and a short stimulation test was performed at 8 p.m. RESULTS: Before Ramadan, cortisol level was higher at 8 a.m. (749.5 +/- 207 nmol/l) than at 8 p.m. (195.18 +/- 79 nmol/l, p<0.001) and the incremental cortisol response to exo genous ACTH was higher at 8 a.m. (peak at 1.167 +/- 46 nmol/l) than at 8 p.m. (peak at 950 nmol/l) (NS). During Ramadan, morning cortisol level (646.3 81 nmol/l) was lower than before Ramadan (NS). The 8 p.m. cortisol level (319 193 nmol/l) increased slightly during Ramadan (p=0.08) but remained lower than the morning level during Ramadan (p=0.001) and before Ramadan (p<0.001). There was not significant difference between cortisol responsiveness to exogenous ACTH at 8 p.m. during Ramadan (peak at 1.102 51 nmol) and at 8 a.m. and 8 p.m. during Ramadan. DISCUSSION: Changes in eating and sleeping schedule was found to reduce morning cortisol level and raise the evening cortisol level. Adrenal sensitiveness to corticotropin stimulation did not appear to be impaired during Ramadan. PMID- 12527849 TI - [Incidence of factors favoring recurrent foot ulcers in diabetic patients]. AB - BACKGROUND: Patients with foot ulcers have a high risk of relapse and amputation. Several studies have reported that 28 to 51% of amputated diabetics will have a second amputation of the lower limb within five years of the first amputation. The purpose of this study was to assess the incidence of factors favoring relapse within two years. MATERIAL AND METHODS: One hundred ten diabetic patients were treated for foot lesions in the Casablanca Ibn Rochd university hospital between 1997 and 2000. Ninety of these patients attended review consultations. RESULTS: There were 42 cases of relapse (46.6%). Male gender predominated in the relapse patients with a sex ratio of 3.2. Mean age at relapse was 55 years; 71.5% of the patients had type 2 diabetes. Lesions observed were neuropathic ulcer (n=23), arterial ulcer (n=6), infected wounds (n=13). Revealing factors were burns and wounds. The main risk factors were neuropathy (n=23, 52%), neuroarteriopathy (n=12, 31%), peur arteriopathy (n=6, 12%). Patients who relapsed (n=42) were significantly different from patients who did not relapse (n=48) for gender, presence or absence of neuropathy, and presence or absence of arteriopathy. DISCUSSION: Secondary preventive measures against these risk factors, medical care, and specialized follow-up were satisfactory in these patients. our findings illustrate the importance of specialized management of diabetic patients with foot lesions. Adequate care of the lesions and preventive measures against risk factors are needed. PMID- 12527851 TI - [Impetigo herpetiformis, a rare manifestation of severe hypocalcemia]. AB - A case of impetigo herpetiformis associated with profound hypocalcemia secondary to idiopathic hypoparathyroidism is reported. This clinical observation and the review of similar cases in the literature recall that this rare and severe skin disease related to psoriasis can be the first manifestation of hypoparathyroidism and responds very well to the calcium reloading. PMID- 12527852 TI - [Three-year follow-up of infraclinical hypothyroidism: course in a cohort of 97 women aged 45 to 70 years attending healthcare screening centers]. AB - Infraclinical hypothyroidism is a recognized entity defined in terms of laboratory results which occurs more readily in women. The appropriateness of thyroid hormone multicentric prospective study initiated in 1997 has established that infraclinical hypothyroidism occurred in about 3% of women over 45 years of age undergoing routine check-ups at eleven health screening centers. Associated clinical signs and laboratory findings were reported. The purpose of the present study was to describe findings in a 3-year follow-up of these women with infraclinical hypothyroidism in order to assess natural history and appropriate care. Ninety-seven women of the 151 women with a TSH level between 4 and 12 mU/l in the 1997 survey were reviewed during the last three months of 2000. Forty three of them had taken thyroid hormone replacement drugs since 1997. About 44% of the women examined still had infraclinical hypothyroidism or developing hypothyroidism. There was not significant difference between treated (33%) and non-treated (55%) patients. Clinical and biological signs associated with infraclinical hypothyroidism at the initial examination (elevated serum cholesterol, abnormal ECG, eyelid edema, recent weight gain) were unchanged at the second examination, both in treated and not treated women. A logistic model including all these factors demonstrated that the probability of prescription of thyroid hormone replacement therapy increased 5.4-fold when the starting TSH level was above 6 mU/l and 3.2-fold if ECG abnormalities were present. Using a statistical model with anti TPO antibodies, a level above 64 U/l increased the probability of treatment 7-fold, with ECG findings being related to probability of treatment. Age, cholesterol level, use to lipid lowering drugs, presence of eyelid edema or weight gain were not associated with replacement therapy in this model. No consistent conclusion concerning the appropriateness of screening for infraclinical hypothyroidism can be drawn from these three-year follow-up findings. Another examination scheduled for 2003 should provide further precision concerning the natural history of infraclinical hypothyroidism as well as long term medical practices and therapeutic impact. PMID- 12527853 TI - [Thiazolidinediones in type 2 diabetes. Role of peroxisome proliferator-activated receptor gamma (PPARgamma)]. AB - Thiazolidinediones (TZDs) form a new class of oral antidiabetic agents. They improve insulin sensitivity and reduce glycemia, lipidemia and insulinemia in patients with type 2 diabetes. Their mechanism is original, since they activate the nuclear receptor Peroxisome Proliferator-Activated Receptor gamma (PPARgamma), altering the expression of genes involved in glucose and lipid homeostasis. Stimulating PPARgamma improves insulin sensitivity via several mechanisms: 1) it raises the expression of GLUT4 glucose transporter; 2) it regulates release of adipocyte-derived signaling factors that affect insulin sensitivity in muscle, and 3) it contributes to a turn-over in adipose tissue, inducing the production of smaller, more insulin sensitive adipocytes. TZDs also affect free fatty acids (FFA) lipotoxicity on islets, improving pancreatic B-cell function. In addition, triglycerides and FFA levels are lowered by TZDs. Two TZDs, rosiglitazone and pioglitazone, have recently obtained the European commercial licence, but their use is restricted to the association with metformin or sulfonylureas. At the moment, they are indicated in type 2 diabetes but could be of interest in a broader array of diseases related to insulin resistance. As for side effects, rosiglitazone and pioglitazone may cause increased plasma volume, edema and dose-related weight gain. TZDs offer an attractive option in the treatment of type 2 diabetes, though it may be too soon to determine if they prevent vascular complications, as do other oral antidiabetic agents. An important issue for the future will be to assess the influence of weight gain in the long time. PMID- 12527854 TI - Efficacy and safety of bromocriptine in the treatment of macroprolactinomas. AB - To assess the effectiveness of bromocriptine (BRC) as primary therapy in reducing the size of PRL-secreting macroadenomas with extra-sellar extension, we conducted a multicenter study in 29 patients without prior radiotherapy. Clinical presentation, response to medical treatment and long term follow-up of 29 patients with macroprolactinoma (pituitary mass more than 10 mm in diameter) were analysed. There were 19 women for 10 men. Mean age was 33 years. An hypogonadism was present in 94% of women vs 57% for men. Headaches and or visual abnormalities were present in 68% of women vs 90% of men. The mean basal serum prolactin level before treatment was 1 501 ng/ml: 202 ng/ml for women (range: 70-478 ng/ml) and 3 870 ng/ml for men (range: 100-20 476 ng/ml) and was correlated to tumoral size. 25 patients were treated with BRC as primary therapy with a mean dose of 9,7 mg/day (13 mg/day in men and 8 mg/day in women). BRC no malized serum PRL levels in all men and in 17/19 women over a mean period of 6 months, ranging from 3 to 72 months in both sexes. Tumor size was reduced by more than 50% in 18/29 patients (62%) with secondary empty sella in 5 patients and by less than 50% in 11 patients. Visual field improved in most of the patients in whom it was initially abnormal. Reduction in size was quite fast in most of patients but slower in some of them. The cumulative radiological response to BRC increased with time. Therefore, it appears that prolonged medical therapy is effective and safe in macroprolactinomas. PMID- 12527855 TI - [Effect of lanreotide on prolactin level in patients with pituitary mixed tumors]. AB - Acromegaly is a disease caused by a pituitary tumor (somatotropinoma) or by ectopic secretion of GH or IGF-1. About 15% of tumors secrete not only GH but PRL as well. Last time a lanreotide and an octreotide (the somatostatine analogues) are useful in the therapy of acromegaly. We observed the influence of the lanreotide on GH and prolactin. We noticed that the lanreotide caused not only serum level reduction of a growth hormone but also prolactine in patients with mixed pituitary tumors. PMID- 12527856 TI - [Acromegaly due to ectopic GH RH secretion by a bronchial carcinoid tumor: a case report]. AB - Ectopic GH RH is a very uncommon cause of acromegaly (<1% of acromegaly). We report the case of an 41 year-old woman with acromegaly due to a GH-RH-secreting bronchial carcinoid tumor. Elevated serum GH (14 ng/ml) was paradoxically stimulated after glucose loading. Magnetic resonance imaging revealed a normal pituitary gland. The thoracic CT demonstrated a voluminous (8 cm) bronchial tumor visualised with octreotide scintigraphy. Acromegaly due to ectopic GH RH secretion was confirmed by very high plasma immunoreactive GH RH level. After removal of the carcinoid tumor, plasmat GH, IGF1 and GH RH levels were normalised. PMID- 12527857 TI - Vitamin D deficiency and severe hyperparathyroidism. AB - The typical manifestations of severe hypercalcemia with osteitis fibrosa cystica have become exceedingly rare. We describe the case of a woman hospitalized for a tibial tumor with functional impotence, leading to a diagnosis of primary hyperparathyroidism (HPT I) associated with profound vitamin D deficiency. This 31-year-old woman was admitted, after two pregnancies complicated by the HELLP syndrome. Preoperative laboratory values were as follows: calcemia 4.05 mmol/l (2.2-2.6); urinary calcium 30 mmol/24 h (1.25-7.5); parathormone (PTH) 1 195 pg/ml (10-60); and 25 OH-vitamin D 13 nmol/l (22-120). Specific MIBI uptake by the tibial lesion oriented the diagnosis towards a brown tumor. After surgical excision of a parathyroid adenoma and the brown tumor (associated with tibial fracture), calcemia fell to 1.55 mmol/l and normalized after three months. Urinary calcium fell to 0.1 mmol/24 h and remained low during the 2 years following surgery. Vitamin D levels rapidly normalized on supplementation (87 nmol/l). PTH levels fell markedly after surgery but remained higher than normal till 2 years after surgery despite normalization of calcemia three months after. Bone repair, estimated by means of bone densitometry, improved from preoperative Z-score values of - 6.54, - 5.20 and - 3.50 in the left femoral neck, right femoral neck and lumbar spine, respectively, to - 0.20, - 1.55 and - 0.28, respectively, one year after surgery. In conclusion, this case illustrates: 1) the severe osseous expression of HPT probably related to vitamin D deficiency; 2) specific MIBI uptake by the bone lesion, orientating the diagnosis towards a brown tumor; 3) the consequences of vitamin D deficiency on postoperative outcome, with transient severe initial hypocalcemia related to bone calcium avidity; 4) a possible link between HPT and the HELLP syndrome. PMID- 12527858 TI - [Report from fact-finding committee aquisition of dosimetry in x-ray CT equipment]. PMID- 12527859 TI - [Evaluation of imaging properties of recent screen-film systems]. PMID- 12527862 TI - [Apparatus engineering of X ray CT equipment (1) : generation and detection of X rays--]. PMID- 12527863 TI - [What I learned by visiting Toshiba]. PMID- 12527865 TI - [An effective period of the use of medical appliances]. PMID- 12527866 TI - [Utility of fixed-quantity analysis in evaluating image noise and resolution in multi-slice CT]. AB - We analyzed the spatial frequency of images generated with multi-slice CT (Z axis) and estimated the definition and noise characteristics. The noise characteristics were calculated with Wiener spectra from multi planar reconstruction (MPR) images, and, in the evaluation of definition characteristics, the modulation transfer function (MTF) was calculated using section sensitivity profile (SSP) with the base method. The Wiener spectra showed a tendency for noise to decrease from a low frequency domain as the effective slice thickness increased. In addition, Wiener spectra were not much different in helical pitch because effective slice thickness became thicker with increases in helical pitch. Examination results suggest that quantitative analysis of the characteristics of image noise and resolution in multi-slice CT images may provide an optical parameter for improving the quality of images in clinical data. PMID- 12527867 TI - [Comparison of contrast resolution between dynamic MRI and dynamic CT in liver scanning]. AB - Dynamic studies employing contrast agents have been used in the detection of hepatocellular carcinoma (HCC). Some studies have indicated that the detectability of small nodules in liver is significantly better with MRI dynamic study than with CT dynamic study. However, the reasons for this are unclear. In an attempt to clarify this difference, we compared contrast resolution using two contrast phantoms made for MRI and CT. Both phantoms were constructed using the same ratios of dilution of the contrast materials Gd-DTPA and iodine. The signal intensity of different dilution ratios of contrast medium were measured in order to compare the signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), and contrast. The results showed that the contrast of phantoms in MRI and CT dynamic studies was almost the same except for certain MRI scan techniques. In terms of SNR and CNR, MRI dynamic study was superior to CT dynamic study. In addition, T(1) relaxation of the contrast agent should be considered for suitable scanning, because the contrast of MRI was affected by scan parameters. PMID- 12527868 TI - [Examination of the validity of RF power determination by auto prescan in T(1) weighted images]. AB - In the sequence of spin echo MRI systems, RF power selects the output value from which the signal intensity of the whole photography volume changes to the maximum as a 90-degree pulse in auto tuning during prescanning by the SIGNA HORIZON LX 1.0 Tesla ver. 8.3. When it consists of two or more organizations like a human body, although the RF power to which signal intensity serves as the maximum is in agreement with the RF power of a 90-degree pulse if it is a uniform phantom, the RF power from which signal intensity reaches the maximum differs for every organization. When we determined RF power by the signal intensity of two or more whole organizations in this experiment, it was found that RF power was strongly influenced by a high signal organization. Moreover, if CNR in two or more organizations is considered, T(1) contrast will be emphasized for the direction of an angle less than 90 degrees. It was possible for the same effect to be acquired by lowering the power of the RF pulse and emphasizing T(1) contrast. PMID- 12527869 TI - [Comparison of the depiction of pancreaticoduodenal arcades and dorsal pancreatic artery, using three-point scale with volume rendering (VR), maximum intensity projection (MIP), and shaded surface display (SSD)]. AB - Preoperative visualization of pancreaticoduodenal arcades and the dorsal pancreatic artery, which supply the pancreatic head and body, is crucial to surgeons, who must guard against ligation or injury during pancreatic surgery. The peripancreatic arteries now can be noninvasively displayed using three dimensional CT angiographic techniques (3D CTA) so that peripancreaticoarterial anatomy can be defined before pancreatic surgery, obviating the need for conventional catheter angiography. We compared the depiction of the pancreaticoduodenal arcades and dorsal pancreatic artery, using volume rendering (VR), maximum intensity projection (MIP), and shaded surface display (SSD) as post-processing techniques. The results of visual grading (using a 3-point scale) indicated that VR was superior to MIP and SSD in the depiction of pancreaticoduodenal arcades and the dorsal pancreatic artery. Unlike the other rendering techniques, VR can also depict relationships between these vessels and the pancreatic parenchyma and adjacent structures. PMID- 12527870 TI - [Report on the 86th Scientific Assembly and Annual Meeting of the Radiological Society of North America: motion artifact caused by heart pulsation observed on an electron beam and a multidetector-row CT]. AB - Recently developed technologies in CT imaging have allowed higher temporal resolution and less motion artifacts caused by heart pulsation. However, complete deletion of motion artifacts has not altogether been accomplished. In interpreting images, differentiation of motion artifacts and true lesion is essential. In this exhibition, the ascending aorta, outer contour of the heart, valve, lung adjacent to the heart, and other structures in the mediastinum are demonstrated focusing on the characteristics of motion artifact. Images obtained with an electron beam CT (Imatron 150XL) and a MDCT (LightSpeed QX/i, GEMS) are comparatively demonstrated. In electron beam CT, the temporal resolution ranged from 50 msec to 2,000 msec. In multidetector-row CT, half reconstruction algorithm (500 msec) and newly developed algorithm (130 msec) can be applied to reduce temporal resolution, although temporal resolution (800msec) of a full scan is fixed. Degree, shape, and site at which motion artifacts arise are analyzed with respect to the temporal resolution on two different CT systems. LEARNING OBJECTIVES: Motion artifacts are correlated with the temporal resolution of the CT scanner. Electron beam and multidetector row CT provides different kinds of solutions to reduce motion artifacts. PMID- 12527871 TI - [Report on the 13th European Congress of Radiology (ECR): Comparison of primary proton beams and secondary charged-particles]. AB - PURPOSE: In the proton radiotherapy, primary proton beams contribute to the absorbed dose and the share of secondary charged-particles is small. The purpose is to discuss about the ratio of average dose of primary proton beams and secondary charged-particles. METHODS: We performed the dosimetry of 70 MeV proton beams in water using ionization chamber. The ratio of average dose for secondary charged-particles in some range shifter was calculated by the dose ratio of primary and scatter. To exclude the influence of lateral secondary charged particles, the ratio of average dose for secondary charged-particles was extrapolated to zero field size of each. RESULTS: The ratio of average dose for secondary charged-particles was extrapolated to zero field size by the bi exponential fit. The ratio of average dose for longitudinal secondary charged particles for zero fields at each depth is almost the same; the different at the shallow depth is decreased. CONCLUSIONS: The secondary charged-particles from collimator is affected by the dose of shallow depth. The influence of lateral secondary charged-particles in water phantom was excluded with the extrapolation method for zero field size in each depth. PMID- 12527872 TI - [Investigation report on causes of radiation overexposure incident at a hospital in Tokyo city area and prevention of similar accidents in radiotherapy by the Japan Association on Radiological Physics]. PMID- 12527873 TI - [Measurement of cerebral hemodynamics using DSC-MRI]. PMID- 12527874 TI - [History of X ray CT equipment - past, present, and the future -]. PMID- 12527875 TI - [Woek on "terminology" in JIRA]. PMID- 12527876 TI - [Request for approval of manufacture (import) of standalone software]. PMID- 12527877 TI - [Method of preventing aliasing artifacts of arms using electromagnetic-wave shield fabric on coronal images of the thorax or abdomen by fast breath-hold MR imaging]. AB - The purpose of this study is to examine an antialiasing method in the breath-hold coronal imaging of the thorax or abdomen. This paper reports a new method of preventing the aliasing artifacts of arms using "Hertz II" as a electromagnetic wave shield fabric (ESF),which is a very thin and flexible special synthetic fabric. In this study, the ESF method was compared with various conventional preventive measures. Temperature changed around the ESF was also measured by using a fiber optic temperature measurement system during the MR scanning procedure. In addition, the ESF method was tried to use for scanning with some volunteers and some patients in clinical examinations. The results of this study showed that the ESF method could efficiently prevent the aliasing of arms in the MR images. Furthermore, safety of the subjects was hardly affected by the ESF method during MR imaging. Therefore, we concluded that the ESF method was an effectual antialiasing method in the imaging of the thorax or abdomen by using the fast breath-hold scanning. PMID- 12527878 TI - [Effects of the volume and shape of voxels on the measurement of phantom volume using three-dimensional magnetic resonance imaging]. AB - Recently, an increasing number of volumetric studies of the human brain have been reported, using three-dimensional magnetic resonance imaging (3D-MRI). To our knowledge, however, there are few investigations on the relation of the volume and shape of voxels which constitute an MR image to the accuracy in volume measurement of an imaged object. The purpose of this study was to evaluate the effect of a different shape of voxel, that is, isotropic or anisotropic, as well as the volume of a voxel on the volume measurement based on the original image data and multiplanar reconstruction (MPR) data, respectively. In the experiment, we repeatedly acquired contiguous sagittal images of a single globe phantom with a known volume under the condition in which the volume and shape of voxels varied, on a 1.5T MR scanner. We used a gradient echo sequence (3D FLASH). The volume of the globe phantom from both original images and MPR ones was measured on workstations employing a semi-automated local thresholding technique. As a result, the smaller volume of voxels tended to give us the more correct measurement, and an isotropic voxel reduced measurement errors as compared to an anisotropic one. Therefore, it is concluded that the setting of voxel with both an isotropic shape and small volume, e.g., a voxel of 1 mm x 1 mm x 1 mm at present, is recommended in order to get a precise volume measurement using 3D MRI. PMID- 12527879 TI - [Evaluation of the retrospective ECG-gated helical scan using half-second multi slice CT: motion phantom study for volumetry]. AB - ECG synchronized technique on multi-slice CT provide the thinner (less 2 mm slice thickness) and faster (0.5sec/rotation) scan than that of the single detector CT and can acquire the coverage of the entire heart volume within one breath-hold. However, temporal resolution of multi-slice CT is insufficient on practical range of heart rate. The purpose of this study was to evaluate the accuracy of volumetry on cardiac function measurement in retrospective ECG-gated helical scan. We discussed the influence of the degradation of image quality and limitation of the heart rate in cardiac function measurement (volumetry) using motion phantom. PMID- 12527880 TI - [The radiation protective devices for interventional procedures using computed tomography]. AB - A scattered dose and a surface dose from phantom measurements during interventional procedures with computed tomography (IVR-CT) were evaluated. To reduce the personnel exposure in IVR-CT, the new protective devices were developed and its effect evaluated. Two radiation protection devices were experimentally made using a lead vinyl sheet with lead equivalent 0.125mmPb. The first device is a lead curtain which shields the space of CT-gantry and phantom for the CT examination. The second device is a lead drape which shields on the phantom surface adjacent to the scanning plane for the CT-fluoroscopy. Scattered dose and phantom surface dose were measured with an abdominal phantom during Cine CT (130 kV, 150 mA, 5 seconds, 10 mm section thickness). They were measured by using ionization chamber dosimeter. They were measured with and without a lead curtain and a lead drape. Scattered dose rate was measured at distance of 50-150 cm from the scanning plane. And, surface dose was measured at distance of 4-21 cm from the scanning plane on the phantom. On operator's standing position, scattered dose rates were from 8.4 to 11.6 micro Gy/sec at CT examination. The lead curtain and the lead drape reduced scattered dose rate at distance of 50 cm from the scanning plane by 66% and 58.3% respectively. Surface dose rate were 118 micro Gy/sec at distance of 5 cm from the scanning plane at CT-fluoroscopy. The lead drape reduced the surface dose by 60.5%. High scattered exposure to personnel may occur during interventional procedures using CT. They were considerably reduced during CT-arteriography by attaching the lead curtain in CT equipment. And they were substantially reduced during CT-fluoroscopy by placing the lead drape adjacent to the scanning plane, in addition, operator's hand would be protected from unnecessary radiation scattered by phantom. It was suggested that the scattered exposure to personnel could be sufficiently reduced by using radiation protection devices in IVR-CT. The radiation protection devices and the CT equipment should be improved or developed based on the radiation protection. PMID- 12527881 TI - [Study of image quality (contrast) and reduction of patient dose by using heavy metal filters]. AB - We studied image quality (contrast) and patient dose reduction using heavy metal filters in lumbar spine and abdomen x-ray examination. Heavy metal filters used in this study are gadolinium, holmium and ytterbium and these combinations. These filters have k-absorption edge in the range from 50 to 70 keV. Image quality and patient dose in 70-90 kV tube voltage with heavy metal filters were compared with 80 kV tube voltage without filter. Image quality was improved in four percent and patient dose could be reduced by 30%. However, tube loading increased from 1.6 to 2.2 times. It was found that the best filter choices gave better image and reduced patient dose compared to without filter. PMID- 12527882 TI - [Depth dose characteristics of electron beams released from a scanning type Racetrack Microtron treatment machine]. AB - The Racetrack Microtron MM50 capable of taking out x-rays and electron beams having a high energy of up to 50 MeV was evaluated by a dosimetry of electron beams in comparison with Microtron MM22. The MM50 flattens the intensity of electron beams by using the beam scanning method while the MM22 utilizes the flattening-filter method. A percentage depth dose (PDD) curve was obtained through the dosimetry of electron beams using a water phantom. As compared with the MM22, the MM50 emits an electron beam that has an energy much closer to the nominal one, that is less contaminated by x-rays, and whose intensity decreases steeply down to near zero on the PDD curve. The MM50 has an electron beam dose distribution that is practically useful since the dose tends to be concentrated on the target volume. PMID- 12527883 TI - [Evaluation of compensation filter in I.I.-DR gastrointestinal examination using image pick-up tube]. AB - We have especially the thing which experiences daily and which digital value does saturate by the difference of patient, and the air content of internal organs, at the time of I.I.-DR gastrointestinal examination. In order to compensate digital value of those parts, the compensation filter by the remote operation formula was examined. The quality of the material of the created filter determined thickness and form from change of digital value when changing thickness, and a dose using aluminum. The image which carried out saturate as a result of analyzing this 500 images decreased to 15% to 3%. Moreover, the total skin dose decreased on the same photography conditions, and was almost equivalent in the time of phototimer use. The part not conventionally materialized as an image for saturation has been improved by using a compensation filter for gastrointestinal examination, and it was enabled to obtain a useful picture clinically. PMID- 12527884 TI - [Management of bio waste by the area monitor system]. PMID- 12527885 TI - ATM is a target for positive regulation by E2F-1. AB - The E2F-1 transcription factor is a critical downstream target of the tumor suppressor, RB. When activated, E2F-1 induces cell proliferation. In addition, deregulation of E2F-1 constitutes an oncogenic stress that can induce apoptosis. The protein kinase ATM is a pivotal mediator of the response to another type of stress, genotoxic stress. In response to ionizing radiation, ATM activates the tumor suppressor p53, a key player in the control of cell growth and viability. We show here that E2F-1 elevates ATM promoter activity and induces an increase in ATM mRNA and protein levels. This is accompanied by an E2F-induced increase in p53 phosphorylation. Expression of the E7 protein of HPV16, which dissociates RB/E2F complexes, also induces the elevation of ATM levels and p53 phosphorylation, implicating endogenous E2F in these phenomena. These data demonstrate that ATM is transcriptionally regulated by E2F-1 and suggest that ATM serves as a novel, ARF-independent functional link between the RB/E2F pathway and p53. PMID- 12527886 TI - Transcription-independent triggering of the extrinsic pathway of apoptosis by human papillomavirus 18 E2 protein. AB - Cervical carcinomas are most frequently associated with human papillomaviruses (HPV), whose E6 and E7 oncogenes products induce cellular immortalization. The papillomavirus E2 protein is a transcription factor, which represses the expression of the viral oncogenes, and activates viral DNA replication during the vegetative viral cycle. This protein is specifically inactivated in HPV18 associated carcinoma cells, suggesting that E2 functions prevent carcinogenic progression. Indeed, ectopic expression of E2 in cervical carcinoma cells strongly inhibits cell proliferation. Here we show that above a threshold level of expression, the E2 protein induces apoptosis, independently of other viral functions. The amino-terminal domain is responsible for this apoptotic activity, but surprisingly with no involvement of its transcriptional functions. The death pathway triggered by E2 relies on activation of the initiator caspase 8, specific of the extrinsic pathway of apoptosis. E2 itself is cleaved by caspases during cell death, providing an example of an apoptotic inducer that is itself a target for caspase processing. The autonomous proapoptotic activity of HPV18 E2 described here may counteract the proliferative functions of viral oncogenes, and renders the inactivation of E2 crucial for carcinogenic progression. PMID- 12527887 TI - The tyrosine kinase Lck is involved in regulation of mitochondrial apoptosis pathways. AB - The induction of apoptosis requires the activation of a highly coordinated signaling network ultimately leading to the activation of caspases. In previous experiments we and others have shown that the tyrosine kinase Lck is required for adequate apoptosis induction in response to ionizing radiation, ceramide incubation and overexpression of the HIV-TAT protein. However, the position of Lck within given apoptotic signaling cascades remains unclear. We therefore aimed to define the role of Lck during radiation-induced apoptosis. Apoptosis induction in response to ionizing radiation, CD95 or TRAIL receptor stimulation was determined in Jurkat T-cells, the Lck-deficient Jurkat clone JCaM1.6- and Lck retransfected JCaM1.6/Lck. No apoptosis, release of cytochrome c, breakdown of the mitochondrial potential were detectable during the first 48 h after irradiation of JCaM1.6 cells. In parallel, no activation of caspase-9, -8 and -3 was detectable. Since mitochondrial apoptosis pathways act within a feedback mechanism during death-receptor-mediated apoptosis, the influence of the Lck defect on CD95/Fas/Apo-1-L or TRAIL-induced apoptosis was also tested. Both stimuli induced apoptosis in Lck-deficient cells. However, the kinetics of apoptosis induction determined by caspase-8, -9 and -3 activation as well as deltapsi(m) breakdown was slowed. We conclude that the Lck deficiency influences early steps during radiation-induced mitochondrial alterations. PMID- 12527888 TI - Altered expression of the RON receptor tyrosine kinase in primary human colorectal adenocarcinomas: generation of different splicing RON variants and their oncogenic potential. AB - The RON receptor tyrosine kinase is a member of the MET proto-oncogene family that has been implicated in regulating motile-invasive phenotypes in certain types of epithelial cancers. The purpose of this study was to determine if RON expression is altered in primary human colorectal adenocarcinomas. Results from immunohistochemical staining showed that RON is highly expressed in the majority of colorectal adenocarcinomas (29/49 cases). Accumulated RON is also constitutively active with autophosphorylation in tyrosine residues. Moreover, three splicing variants of RON, namely RONdelta165, RONdelta160, and RONdelta155 were detected and cloned from two primary colon cancer samples. These RON variants were generated by deletions in different regions in extracellular domains of the RON beta chain. Functional studies showed that expression of RONdelta160 or RONdelta155 in Martin-Darby canine kidney cells resulted in increased cell dissociation (scatter-like activity). RON variants, RONdelta160 and RONdelta155, also exerted the ability to induce multiple focus formation and sustain anchorage-independent growth of transfected NIH3T3 cells. Moreover, NIH3T3 cells expressing RONdelta160 or RONdelta155 formed tumors in athymic nude mice and colonized in the lungs. These data suggest that RON expression is altered in certain primary colon cancers. Abnormal accumulation of RON variants may play a role in the progression of certain colorectal cancers in vivo. PMID- 12527890 TI - Differential requirement of EGF receptor and its tyrosine kinase for AP-1 transactivation induced by EGF and TPA. AB - The transcription factor activator protein-1 (AP-1) has been implicated in a large variety of biological processes including cell differentiation, proliferation, apoptosis and oncogenic transformation. It is thought that the 12 O-tetradecanoylphorbol-13-acetate (TPA)-induced AP-1 activity is because of the activation of the PKC/MAPK/AP-1 pathway, although the detailed molecular mechanism has not been fully characterized. The tyrosine kinases of epidermal growth factor receptor (EGFR) lie at the head of a complex of signal transduction cascade that modulates cell proliferation, survival, adhesion, migration and differentiation. Currently, little is known about whether EGFR or its tyrosine kinase is necessary for TPA-induced AP-1 activation. In the present study, we investigated this issue using a well-characterized mouse fibroblast B82 cell line, which is devoid of the EGFR, and its stable transfectants with either wild type EGFR (B82L) or tyrosine kinase-deficient EGFR (mutation at Lys-721) (B82M721). We demonstrated that the TPA or epidermal growth factor (EGF) induced AP-1 activation in the B82L cells that express wild-type EGFR, but not in the B82 cell, whereas autophosphorylation at tyrosine(1173) of EGFR in B82L cells was only induced by EGF, but not TPA. The expression of tyrosine kinase-deficient EGFR (mutation at Lys-721) (B82M721) resulted in deficiency of AP-1 induction in cellular response to EGF, while TPA treatment led to fully AP-1 activation. Furthermore, the mutation at Lys-721 of EGFR resulted in impairing of EGFR autophosphorylation at tyrosine(1173) induced by EGF. Based on these results, we conclude that TPA-induced AP-1 activation requires the basal level-EGFR protein, but not EGFR tyrosine kinase and EGFR autophosphorylation at tyrosine(1173), whereas both EGFR tyrosine kinase and EGFR autophosphorylation at Y(1173) play a critical role in EGF-induced AP-1 activation. PMID- 12527889 TI - Fusion proteins of retinoid receptors antagonize TGF-beta-induced growth inhibition of lung epithelial cells. AB - Transforming growth factor-beta1 (TGF-beta) is a growth factor that has multiple functions including potent inhibition of cell growth. TGF-beta signals by binding to its cell surface serine/threonine kinase receptors, which in turn phosphorylate downstream signal transducers, Smad2 and Smad3. Phosphorylated Smad2 and Smad3, together with Smad4, enter the nucleus and associate with various transcription factors. This complex of transcription factors regulates transcription of a diverse group of genes, leading to growth arrest at G1 phase. Through a functional expression cloning approach, a gag-retinoid X receptor beta (gag-RXRbeta) fusion protein was found to antagonize TGF-beta-induced growth inhibition of mink lung epithelial cells and the fusion between gag and RXRbeta is essential for resistance to the growth inhibition. Like gag-RXRbeta, the oncogenic PLZF-RARalpha fusion protein also antagonizes TGF-beta-induced growth inhibition, and the fusion between PLZF and RARalpha is essential for resistance to TGF-beta. Moreover, TGF-beta and retinoic acid (RA) cooperatively induce growth inhibition as well as transcription of the p15(ink4b) gene, while PLZF RARalpha represses TGF-beta-induced expression of the p15(ink4b) gene. Together, these results suggest that the TGF-beta and RA pathways cooperate to inhibit cell growth and that PLZF-RARalpha -mediated resistance to TGF-beta may facilitate the development of the PLZF-RARalpha-induced leukemia. PMID- 12527891 TI - Protein kinase CK2 regulates CDC25B phosphatase activity. AB - Human dual-specificity phosphatases CDC25 (A, B and C) play an important role in the control of cell cycle progression by activating the cyclin-dependent kinases (CDKs). Regulation of these phosphatases during the cell cycle involves post translational modifications such as phosphorylation and protein-protein interactions. Given the suspected involvement of the protein kinase CK2 at the G2/M transition, we have investigated its effects on the CDC25B phosphatase. We show that in vitro CK2 phosphorylates CDC25B, but not CDC25C. Mass spectrometry analysis demonstrates that at least two serine residues, Ser-186 and Ser-187, are phosphorylated in vivo. We also report that CDC25B interacts with CK2, and this interaction, mediated by the CK2beta regulatory subunit, involves domains that are located within the first 55 amino acids of CK2beta and between amino acids 122 and 200 on CDC25B. This association was confirmed in vivo, in Sf9 insect cells and in U(2)OS human cells expressing an HA epitope-tagged CDC25B. Finally, we demonstrate that phosphorylation of CDC25B by protein kinase CK2 increases the catalytic activity of the phosphatase in vitro as well as in vivo. We discuss the possibility that CDC25B phosphorylation by CK2 could play a role in the regulation of the activity of CDC25B as a starter of mitosis. PMID- 12527892 TI - p53-associated 3'-->5' exonuclease activity in nuclear and cytoplasmic compartments of cells. AB - The tumor suppressor protein p53 plays an important role in maintenance of the genomic integrity of cells. p53 possesses an intrinsic 3'-->5' exonuclease activity. p53 was found in the nucleus and in the cytoplasm of the cell. In order to evaluate the subcellular location and extent of p53-associated 3'--> 5' exonuclease activity, we established an in vitro experimental system of cell lines with different nuclear/cytoplasmic distribution of p53. Nuclear and cytoplasmic extracts obtained from LCC2 cells (expressing a high level of cytoplasmic wild-type p53), MCF-7 cells (expressing a high level of wild-type nuclear p53), MDA cells (expressing mutant p53) and H1299 cells (p53-null) were subjected to the analysis of exonuclease activity. Interestingly, 3'-->5' exonuclease was predominantly cytoplasmic; the nuclear extracts derived from all cell lines tested, exerted a low level of exonuclease activity. Cytoplasmic extracts of LCC2 cells, with a high level of wild-type p53, showed an enhanced exonuclease activity in comparison to those expressing either a low level of wild type p53 (in MCF-7 cells) or the mutant p53 (in MDA cells). Evidence that exonuclease function detected in cytoplasmic extracts is attributed to the p53 is supported by several facts: First, this activity closely parallels with levels and status of endogenous cytoplasmic p53. Second, immunoprecipitation of p53 from cytoplasmic extracts of LCC2 cells markedly reduced the exonuclease activity. Third, the observed 3'-->5' exonuclease in cytoplasmic fraction of LCC2 cells displays identical biochemical properties characteristic of recombinant wild-type p53. The biochemical functions include: (a) substrate specificity; exonuclease hydrolyzes single-stranded DNA in preference to double-stranded DNA and RNA/DNA template-primers, (b) efficient excision of 3'-terminal mispairs from DNA/DNA and RNA/DNA substrates, (c) the preferential excision of purine-purine mispairs over purine-pyrimidine mispairs and (d) functional interaction with exonuclease deficient DNA polymerase, for example, murine leukemia virus reverse transcriptase (representing a relatively low fidelity enzyme), thus enhancing the fidelity of DNA synthesis by excision of mismatched nucleotides from the nascent DNA strand. Taken together, the data demonstrate that wild-type p53 in cytoplasm, in its noninduced state, is functional; it displays intrinsic 3'-->5' exonuclease activity. The possible role of p53-associated 3'-->5' exonuclease activity in DNA repair in nucleus and cytoplasm is discussed. PMID- 12527893 TI - Ras-mediated apoptosis of PC CL 3 rat thyroid cells induced by RET/PTC oncogenes. AB - RET gene rearrangements, which generate chimeric RET/PTC oncogenes, are early events in the evolution of thyroid papillary carcinomas. Expression of RET/PTC oncogenes promotes neoplastic transformation of cultured thyroid cells and of thyroid glands in transgenic mice. Notwithstanding these oncogenic effects, we have found that the expression of two RET/PTC oncogenes (H4-RET and RFG-RET) induces apoptosis of rat thyroid PC CL 3 cells. Promotion of thyroid cell death depends on the kinase activity of RET/PTC and on the phosphorylation of a tyrosine residue (tyrosine 1062) that maps in the carboxy-terminus of the RET protein. Tyrosine 1062 is essential for RET/PTC-mediated activation of the Ras/ERK pathway. Inhibition of Ras/ERK by a dominant negative Ras or by the MEKI inhibitor, PD98059, obstructed RET/PTC-mediated apoptosis. We also show that signals transmitted by tyrosine 1062 mediate proapoptotic events like Bcl-2 down regulation and Bax upregulation, and that adoptive overexpression of Bcl-2 overcomes RET/PTC-induced apoptosis. Thus, gene rearrangements that generate RET/PTC oncogenes subvert RET function by converting it into a chronically active kinase that is constitutively phosphorylated on tyrosine 1062. In turn, Y1062 phosphorylation transmits not only mitogenic but also proapoptotic signals to thyroid cells. PMID- 12527894 TI - IL-8 expression and its possible relationship with estrogen-receptor-negative status of breast cancer cells. AB - Estrogen-receptor (ER) status is an important parameter in breast cancer management as ER-positive breast cancers have a better prognosis than ER-negative tumors. This difference comes essentially from the lower aggressiveness and invasiveness of ER-positive tumors. Here, we demonstrate, that interleukin-8 (IL 8) was clearly overexpressed in most ER-negative breast, ovary cell lines and breast tumor samples tested, whereas no significant IL-8 level could be detected in ER-positive breast or ovarian cell lines. We have also cloned human IL-8 from ER-negative MDA-MB-231 cells, and we show that IL-8 produced by breast cancer cells is identical to monocyte-derived IL-8. Interestingly, the invasion potential of ER-negative breast cancer cells is associated at least in part with expression of IL-8, but not with IL-8 receptor levels. Moreover, IL-8 increases the invasiveness of ER-positive breast cancer cells by two fold, thus confirming the invasion-promoting role of IL-8. On the other hand, exogenous expression of estrogen receptors in ER-negative cells led to a decrease of IL-8 levels. In summary, our data show that IL-8 expression is negatively linked to ER status of breast and ovarian cancer cells. We also support the idea that IL-8 expression is associated with a higher invasiveness potential of cancer cells in vitro, which suggests that IL-8 could be a novel marker of tumor aggressiveness. PMID- 12527895 TI - Cloning and characterization of human Src-like adaptor protein 2 and a novel splice isoform, SLAP-2-v. AB - Src-like adaptor protein 2 (SLAP-2) is a recently characterized adaptor protein bearing sequence and structural similarity to the Src-like adaptor protein (SLAP). SLAP-2 expression is hematopoietic-specific, and it has been demonstrated to function as a negative regulator of T-cell antigen receptor (TCR)-mediated signalling by virtue of its interaction with c-Cbl. Here we report the cloning of a cDNA encoding the human homologue of SLAP-2, as well as the genomic structure of the human SLAP-2 gene. Similar to its murine counterpart, two human SLAP-2 protein isoforms exist because of alternative translation initiation, and SLAP-2 protein expression is observed in a variety of hematopoietic cell lines of both lymphoid and myeloid lineages. The human SLAP-2 gene is located on chromosome 20q, and the SLAP-2 coding region consists of seven exons. Concurrent with the cloning of the full-length SLAP-2 cDNA, a unique cDNA encoding an alternatively spliced SLAP-2 isoform has been identified, and designated as SLAP-2-v. The SLAP 2-v transcript encodes a putative protein of 210 amino acids that lacks the c-Cbl interaction region, and consequently is impaired in its ability to both bind to c Cbl, and inhibit TCR signalling relative to SLAP-2. PMID- 12527896 TI - Methylation-associated silencing of heparan sulfate D-glucosaminyl 3-O sulfotransferase-2 (3-OST-2) in human breast, colon, lung and pancreatic cancers. AB - Aberrant CpG methylations play important roles in cancer development and progression. In this study, aberrant methylations in human breast cancer were searched for using methylation-sensitive representational difference analysis (MS RDA). A CpG island (CGI) in the 5' region of the heparan sulfate D-glucosaminyl 3 O-sulfotransferase-2 (3-OST-2) gene was found to be hypermethylated, while its exon 2 was hypomethylated. In seven breast cancer cell lines, hypermethylation of the 5' region and loss of 3-OST-2 expression were observed. Treatment with a demethylating agent, 5-aza-2'-deoxycytidine, removed the methylation of the CGI in the 5' region and restored its expression, demonstrating silencing of the 3 OST-2 gene. Methylation-specific PCR (MSP) analysis in 85 primary breast cancers showed that the hypermethylation of the CGI in the 5' region was present in 75 (88%) of them. Quantitative reverse transcriptase-PCR (RT-PCR) analysis in 37 primary breast cancers showed that the average expression level was decreased in them. Further, MSP analysis in primary colon, lung and pancreatic cancers showed that hypermethylation of the CGI in the 5' region was present in the colon (8/10, 80%), lung (7/10, 70%) and pancreatic (10/10, 100%) cancers. These results showed that silencing of 3-OST-2 was present in a wide range of human cancers. The 3-OST 2 gene encodes an enzyme involved in the final modification step of heparan sulfate proteoglycans (HSPGs), and its silencing is expected to result in abnormal modification of HSPGs and abnormal signal transduction. From the high incidence, silencing of the 3-OST-2 gene is expected to have high diagnostic, and potentially therapeutic, values. PMID- 12527897 TI - Identification of a 0.05). However, the CD15 positive group appeared to show a minor trend toward less positivity for EMA (44% versus 72%), ALK protein (22% versus 51%), and CD45RO (33.3% versus 83.3%, p=0.07), when compared to the typical CD15-negative neoplasms. In summary, although the co-expression of CD30 and CD15 is typical for classical HL, it may be also present in a subset of peripheral T-cell neoplasms including ALK-positive anaplastic large cell lymphoma. Combined and sensible use of morphology and a broad immunophenotypic panel in cases with limited material and/or those with overlapping histologic patterns will best discriminate between HL and ALCL. It is incumbent upon the pathologist to distinguish between these two clinicopathologic entities, since treatment options and clinical outcomes differ. PMID- 12527930 TI - High-throughput genotyping by DHPLC of the dihydropyrimidine dehydrogenase gene implicated in (fluoro)pyrimidine catabolism. AB - Dihydropyrimidine dehydrogenase (DPD) is the first and rate-limiting enzyme in the degradation of pyrimidines and pyrimidine base analogs including the anticancer drugs 5-fluorouracil (5-FU) and Xeloda. A decreased DPD enzyme activity has been described in cancer patients experiencing severe and life threatening toxicity after 5-FU treatment and distinct sequence variants in the DPD gene (DPYD) have been associated with impaired enzyme function. The most prominent mutation in the DPD deficient patient group, a mutation in the splicing donor consensus sequence of intron 14, IVS14+1g>a, resulting in a truncated protein, has been observed in the Caucasian population at frequencies as high as 0.91%-0.94%. This underlines the need for a test system for DPYD mutations in patients undergoing chemotherapy with 5-FU or with Xeloda. To set up a fast and sensitive method to identify variant DPYD alleles, we analyzed 50 healthy individuals by denaturing high performance liquid chromatography (DHPLC). A primer set spanning the whole coding region and the exon-intron boundaries of DPYD was used. In addition, a cDNA-based assay was developed to rapidly identify the 165 base pair deletion in the corresponding RNA of IVS14+1g>a mutation carriers. The optimal mutation detection was elaborated for each of the PCR fragments. DHPLC analysis detected 5 different genetic alterations occurring in the coding region of the gene, as well as 10 intronic sequence variants, respectively. In conclusion, high-throughput screening for DPYD variants by DHPLC may be a reliable tool in the investigation of the molecular basis of DPD deficiency. Furthermore, it will help to identify patients at risk for toxic side effects upon chemotherapy using 5-FU and will facilitate individual treatment of patients. PMID- 12527931 TI - Isolation and characterisation of vascular endothelial growth factor-165 specific scFv fragments by phage display. AB - Vascular endothelial growth factor (VEGF) is a multifunctional cytokine which plays a major role in angiogenesis. Alternative splicing causes the production of several different isoforms (VEGF-A 121, 145, 165, 189, 206). VEGF is essential for tumor angiogenesis and several studies have correlated elevated VEGF levels with tumor stage, metastases and progression. Antibody phage display was employed to isolate two scFv antibody fragments, D8 and F10, with specificity for the VEGF165 isoform. It was shown by ELISA and competitive immunohistochemistry that each clone bound to VEGF165 but not VEGF121. Immunohistochemistry with D8 and F10 on colorectal carcinoma and adenoma sections revealed positive staining similar to that shown by a polyclonal VEGF antibody. The scFv antibody fragments, D8 and F10, will be useful in specifically detecting circulating VEGF165 in cancer patients as most studies to date have quantified the total level of circulating VEGF (121 and 165). These reagents will allow further elucidation of the role of VEGF in tumor angiogenesis. PMID- 12527932 TI - Immunodetection and molecular forms of plasma vascular endothelial growth factor C. AB - Vascular endothelial growth factor (VEGF)-C is a member of the VEGF family. VEGF C is involved in developmental lymphangiogenesis and may be important in pathological lymphangiogenesis, lymphatic invasion and metastasis in carcinoma. We describe the development of an indirect enzyme-linked immunosorbent (ELISA) assay for the quantification of VEGF-C in plasma. Capture of VEGF-C was achieved using goat anti-human VEGF-C antibody, followed by detection with rabbit anti human VEGF-C antibody. The sensitivity of the assay was amplified using the biotin-avidin and enhanced chemiluminescence (ECL) systems. The assay was highly sensitive and reproducible with a detection range of 0.4-100 U/ml and the intra- and inter-assay variations were less than 8%. Substitutional tests demonstrated that the assay was specific for VEGF-C with no cross-reaction with VEGF-A or VEGF D. Practical application of the assay was evaluated in 41 colorectal cancer patients and 31 controls. Median plasma levels of VEGF-C were 35.0 U/ml (range: 17.4-75.9 U/ml) in colorectal cancer patients in contrast to 11.5 U/ml (range: 5.4-21.5 U/ml) in controls (p<0.001). Moreover, VEGF-C levels tended to be elevated in patients with advanced disease compared to early disease, but this was not statistically significant owing to a relatively small number of patients in each group. Immunoprecipitation and immunoblotting confirmed detection of VEGF C in plasma and revealed that two forms of VEGF-C were present in the plasma corresponding to approximately 40 and approximately 80 kDa. The measurement of plasma VEGF-C offers opportunities to explore clinical applications in the management of malignancy, in particular in the prediction of lymphatic spread and in other lymphangiogenesis-related diseases. PMID- 12527933 TI - Immuno-gene therapy with adenoviruses expressing fms-like tyrosine kinase 3 ligand and CD40 ligand for mouse hepatoma cells in vivo. AB - Introduction of genes encoding immuno-stimulatory cytokines into cancer cells is known to enhance antitumor immunity. CD40 ligand (CD40L, CD154) and fms-like tyrosine kinase 3 ligand (Flt3L) are recently identified cytokines, which have been demonstrated to stimulate antitumor immunity in several cancer models. However little is known about antitumor activity of Ftl3L and CD40L against hepatocellular carcinoma (HCC). In the present study, we constructed replication defective adenoviruses expressing Flt3L and CD40L and examined their therapeutic efficacy on mouse HCC, MH134 cells. Subcutaneous injection of MH134 cells genetically engineered to express Flt3L and/or CD40L developed tumors in all the syngeneic immunocompetent mice, but tumor growth was significantly delayed as compared to control mice. Partial inhibition of this antitumor effect in athymic nude mice suggests that both innate and adaptive immunity appear to play a role. It was shown by immunodepletion of NK cells with anti-asialo-GM1 antibody that the effector cells involved in innate immunity are NK cells. In a therapeutic setting, however, injection of adenovirus expressing Flt3L or CD40L into pre established MH134 tumors exhibited no efficacy. These data demonstrate that Flt3L and CD40L induce significant, but only weak, antitumor immunity against MH134 cells presumably through both innate and adaptive immunity. Our results suggest that immuno-gene therapy with Flt3L and CD40L may need adjuvant modalities to achieve strong immune response. PMID- 12527934 TI - The usefulness of digital subtraction imaging with Levovist in the diagnosis of focal hepatic tumors. AB - This study was designed to evaluate digital subtraction imaging (DSI) with Levovist in the diagnosis of hepatocellular carcinoma (HCC), metastatic hepatocellular carcinoma, and hepatic hemangioma. The subjects in this study were 70 patients with 76 nodules of hepatic tumors (48 nodules in 46 cases of hepatocellular carcinoma, 20 nodules in 16 cases of metastatic hepatocellular carcinoma, and 8 nodules in 8 cases of hepatic hemangioma). Contrast enhancement of tumors acquired in the early, portal, and late phases with DSI were compared to classify the tumors. DSI of HCC showed contrast enhancement of 40 nodules (82.2%). High contrast enhancement in the early phase, which was maintained in the portal phase, changed to images with no contrast enhancement with partial persistence of contrast enhancement in the late phase. DSI of metastatic hepatic carcinomas demonstrated contrast enhancement of tumor of 18 nodules (90%) to a high degree in the early and portal phases, which changed to images with no contrast enhancement in the late phases. DSI of hepatic hemangioma maintained high contrast enhancement on tumor margins of 5 nodules (62.5%) and on the entire tumor of 3 nodules (37.5%) in the early, portal, and late phases. DSI of hepatic tumors (hepatocellular carcinoma, metastatic hepatocellular carcinoma, and hepatic hemangioma) provided characteristic findings of contrast enhancement in the early, portal, and late phases, and contribute to differential diagnosis. PMID- 12527935 TI - Regulation of dihydropyrimidine dehydrogenase gene expression in regenerating mouse liver. AB - 5-fluorouracil (5-FU) is one of the most commonly used agents in treatment of the cancer. The administered 5-FU is degraded mainly in the liver by dihydropyrimidine dehydrogenase (DPD), which is the initial rate-limiting enzyme in the catabolic pathway of pyrimidine. This enzyme activity in the tumors has been shown to correlate with the effectiveness of 5-FU against tumors. Therefore, to understand the regulation of DPD gene expression is critical in cancer chemotherapy. Since several studies suggested correlation of DPD activity with the cell proliferation rate we studied the DPD gene expression during liver regeneration. DPD enzyme activity decreased rapidly [24 h after partial hepatectomy (PH): 57% of the control] and remained low until 72 h after PH. Protein content also decreased after PH, however, the lowest level (43.2+/-6.3% of control) was reached only 48 h after PH. The DPD mRNA decreased rapidly to about 20% of control 24 h after PH and then recovered to the control level 72 h after PH. The present results indicate that the DPD gene expression is regulated first at the mRNA level when the hepatocytes enter the cell cycle. The protein content of DPD changed in proportion to the level of DPD mRNA with a 24-h lag, suggesting very little regulation at the translational level. There was a discrepancy between the DPD enzyme activity and the protein content, 24 and 72 h after PH, suggesting that the enzyme activity is modulated also by modification of the protein, and the cell proliferation rate is one of the factors that influences the modification. PMID- 12527936 TI - Role of RLIP76 in lung cancer doxorubicin resistance: I. The ATPase activity of RLIP76 correlates with doxorubicin and 4-hydroxynonenal resistance in lung cancer cells. AB - RLIP76 functions as an ATP-dependent transporter of amphiphilic chemotherapeutic drugs such as doxorubicin (DOX, adriamycin), as well as of glutathione-conjugates of endogenous electrophilic toxins such as 4-hydroxynonenal (4HNE). RLIP76 couples transport and ATP-hydrolysis with a 1:1 stoichiometry, making the ATPase activity of RLIP76 an excellent surrogate for its transport activity. Present studies were performed to determine the relationship of the RLIP76 ATPase activity with DOX and 4HNE resistance in a panel of 13 native human lung cancer cell lines. RLIP76 was purified from each cell line and homogeneity demonstrated by SDS-PAGE and amino acid composition analysis. Anti-RLIP76 antibodies were shown by Ouchterlony double immunodiffusion tests to be non-cross-reactive with any other proteins including P-glycoprotein (Pgp) or multidrug resistance associated protein (MRP). These antibodies completely immunoprecipitated ATPase activity of purified RLIP76 fractions, further confirming homogeneity of purified RLIP76. RLIP76 ATPase purified from NSCLC cell lines was about 2-fold more active than that from SCLC in the absence of the stimulator dinitrophenyl S-glutathione (206+/-47, n=7 vs. 94+/-22, n=6, nmol/min/mg protein, respectively), or in its presence (340+/-60, n=7 vs. 186+/-32, n=6, nmol/min/mg; p<0.01). Partial tryptic digest revealed a 44 kDa internal fragment of RLIP76 beginning at Thr-294 in NSCLC cell lines. This fragment was absent from all SCLC, suggesting the possibility that the activity of RLIP76 in SCLC and NSCLC is differentially regulated through post-translational modifications. Taken together, these findings suggest that RLIP76 activity is a general determinant of 4HNE and DOX resistance, and that its activity contributes to the drug-resistant phenotype of NSCLC. PMID- 12527937 TI - Monensin-mediated growth inhibition of SNU-C1 colon cancer cells via cell cycle arrest and apoptosis. AB - Previously, we showed that monensin, Na+ ionophore, potently inhibited the growth of acute myelogenous leukemia and lymphoma cells. Here, we demonstrate that monensin inhibited the proliferation of solid tumor cells with IC50 of about 2.5 micro M. Monensin induced a G1 or a G2-M phase arrest in these cells. When we examined the effects of this drug on SNU-C1 cells, monensin decreased the levels of CDK2, CDK4, CDK6, cyclin D1 and cyclin A proteins. While p27 was increased by monensin, p21 was not. In addition, monensin markedly enhanced the binding of p27 with CDK2, CDK4 and CDK6. Furthermore, the activities of CDK2-, CDK4- and CDK6 associated kinase were reduced in association with hypophosphorylation of Rb protein. Monensin also induced apoptosis in solid tumor cells. Apoptotic process of SNU-C1 cells was associated with the changes of Bax, caspase-3 and mitochondria transmembrane potential (deltapsim). Taken together, these results demonstrated for the first time that monensin inhibited the growth of solid tumor cells, especially SNU-C1 cells, via cell cycle arrest and apoptosis. PMID- 12527938 TI - Evaluation of the chemosensitivity of head and neck cancer cells based on the diverse function of mutated-p53. AB - Pre-therapeutic evaluation of p53 gene is very important for treating patients with head and neck cancer. However, the analysis for p53 gene has generally been done by immunohistochemistry, polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) and direct sequencing. Functional analysis system for p53 transcriptional activity in mammalian cells is now required. We developed a functional analysis system for p53 transcriptional activity in cancer cells. We used two human head and neck cancer cell lines harboring mutated p53 gene, HSG (Asn30Ser) and TYS (Asp281His), and a human osteosarcoma cell line, Saos-2 as a control. We transfected these cells with luciferase reporter plasmids containing promoter sequence of p53 target genes (p21waf1, BAX, MDM2, p53AIP1 or PUMA). After treating the cells with chemotherapeutic drugs, alteration of the luciferase activity was measured. In HSG cells, none of the target gene promoters was activated by treatment with chemotherapeutic drugs. In TYS cells, p21waf1 promoter was markedly activated by treatment with chemotherapeutic drugs, but Bax and p53AIP1 promoter was not activated. This type of mutated-p53 in TYS cells prevents cell death from DNA damage, and probably accumulates genetic alterations and accelerates the malignant progression of the cells by DNA damaging therapy. Thus, analysis for the diverse function of mutated-p53 may help to determine the therapeutic strategy, especially for chemotherapy and radiation in the individual patients with head and neck cancer. PMID- 12527939 TI - Tie2/Tek expression in breast carcinoma: correlations of immunohistochemical assays and long-term follow-up in a series of 909 patients. AB - The degree of angiogenesis in breast cancer has previously been shown to be an indicator of prognosis, and tumor microvasculature is at present a candidate target for new antiangiogenic therapies. Tie2/tek receptor tyrosine kinase is a novel marker of microvasculature of solid tumors that appears to play a key role in the angiogenesis process in breast cancer. However the prognostic significance of Tie2 has never been demonstrated in this neoplasm. In order to establish the prognostic value of Tie2 in breast carcinoma, we investigated Tie2 expression in a large series of patients and correlated it with long-term follow-up. Tie2 expression was investigated using immunohistochemical assays with a polyclonal antibody on frozen sections in a series of 909 patients, and was correlated with long-term (median, 11.3 years) follow-up. Univariate (Kaplan-Meier) analysis showed that a large Tie2 positive tumor surface (cut off = 7%) was significantly correlated with poor overall survival (p=0.025). Tie2 expression correlated with high metastasis risk among all patients (p=0.00067) and among node negative ones as well (p=0.01). Tie2 immuno-expression was also significantly predictive of relapse in all patients (p=0.003) and in the node negative subgroup (p=0.02). In multivariate analysis (Cox model) Tie2 immunodetection was identified as an independent prognostic indicator. Our results suggest that Tie2 immunohistochemical expression exhibits practical clinical relevance in terms of prognostic prediction. Tie2 expression permits identification of poor outcome patients, in particular node negative ones with high risk of metastasis and relapse. Tie2 immunodetection may further be considered as a potential tool for selecting patients who could benefit in the future from specific antiangiogenic therapy interfering with Tie2 pathway. PMID- 12527940 TI - Further aspects on cellular and cordless telephones and brain tumours. AB - We included in a case-control study on brain tumours and mobile and cordless telephones 1,617 patients aged 20-80 years of both sexes diagnosed during January 1, 1997 to June 30, 2000. They were alive at the study time and had histopathology verified brain tumour. One matched control to each case was selected from the Swedish Population Register. The study area was the Uppsala Orebro, Stockholm, Linkoping and Goteborg medical regions of Sweden. Exposure was assessed by a questionnaire that was answered by 1,429 (88%) cases and 1,470 (91%) controls. In total use of analogue cellular telephones gave an increased risk with odds ratio (OR)=1.3, 95% confidence interval (CI)=1.04-1.6, whereas digital and cordless phones did not overall increase the risk significantly. Ipsilateral use of analogue phones gave OR=1.7, 95% CI=1.2-2.3, digital phones OR=1.3, 95% CI=1.02-1.8 and cordless phones OR=1.2, 95% CI=0.9-1.6. The risk for ipsilateral use was significantly increased for astrocytoma for all studied phone types, analogue phones OR=1.8,95% CI=1.1-3.2, digital phones OR=1.8, 95% CI=1.1 2.8, cordless phones OR=1.8, 95% CI=1.1-2.9. Use of a telephone on the opposite side of the brain was not associated with a significantly increased risk for brain tumours. Regarding anatomical area of the tumour and exposure to microwaves, the risk was increased for tumours located in the temporal area on the same side of the brain that was used during phone calls, significantly so for analogue cellular telephones OR=2.3, 95% CI=1.2-4.1. For acoustic neurinoma OR=4.4, 95% CI=2.1-9.2 was calculated among analogue cellular telephone users. When duration of use was analysed as a continuous variable in the total material, the risk increased per year for analogue phones with OR=1.04, 95% CI=1.01-1.08. For astrocytoma and ipsilateral use the trend was for analogue phones OR=1.10, 95% CI=1.02-1.19, digital phones OR=1.11, 95% CI=1.01-1.22, and cordless phones OR=1.09, 95% CI=1.01-1.19. There was a tendency of a shorter tumour induction period for ipsilateral exposure to microwaves than for contralateral, which may indicate a tumour promotor effect. PMID- 12527941 TI - Differential expressions of cyclin-dependent kinase inhibitors (p27 and p21) and their relation to p53 and Ki-67 in oral squamous tumorigenesis. AB - The cyclin-dependent kinase inhibitors p27Kip1 and p21WAF1/Cip1 play important roles in cell-cycle regulation. Although alterations of these genes have been linked to tumorigenesis of several human carcinomas, their involvement in head and neck squamous tumorigenesis is rarely investigated. To determine the role of these genes in the evolution of squamous carcinoma of the head and neck we evaluated their protein expression by immunohistochemistry in non-dysplastic squamous epithelium, premalignant lesions and oral squamous carcinomas. The p53 gene and Ki-67 expressions were correlated with traditional clinicopathologic variables. Our study shows that in histologically non-dysplastic squamous epithelium, p27 expression was noted mainly in superficial differentiated cells, whereas p21, p53 and Ki-67 staining was observed in basal and suprabasal cells. In dysplasia, divergent expression between p27 and p21 was observed: p27 precipitously decreased and p21, p53, and Ki-67 increased with histologic progression. In squamous carcinomas, p27 was mainly expressed in well differentiated tumor cell nests, while the expressions of p21, p53, and Ki-67 were variable in the poorly differentiated tumor areas. A significant inverse relationship between p27 expression and those of p21, p53, and Ki-67 was observed, but no significant association between any of these markers and clinicopathologic factors was noted in this cohort. Our study indicates that: i) down-regulation of p27 and up-regulation of p21 are associated with early progression of HNSC, ii) p21 expression correlates positively with proliferation while p27 correlates positively with cell differentiation and iii) concurrent p27 and p21 expression analysis may allow for better assessment of HNSC progression. PMID- 12527942 TI - Induction of caspase-dependent and -independent apoptosis in response to methionine restriction. AB - Tumor cells are more sensitive to methionine restriction than normal tissues, a phenomenon known as methionine auxotrophy. Previous studies have demonstrated that methionine restriction causes tumor cell growth arrest and eventually apoptosis. The current studies were undertaken to elucidate the molecular pathways leading to apoptosis induced by methionine restriction. We found that methionine restriction induced formation of oligonucleosomal DNA fragment and cytochrome c release from mitochondria in methionine-dependent PC3 and Hela cells. Methionine restriction also led to cleavage and activation of initiator and effector caspases in Hela cells but not PC3 cells. Furthermore, methionine restriction resulted in cleavage of BID and reduction in Bcl-2 levels in both cell lines. These data suggest that apoptosis induced by methionine restriction is mitochondria-dependent. Methionine restriction induced caspase-independent cell death in PC3 cells, whereas it stimulated caspase-dependent cell death in Hela cells. Cleavage of BID and decreased expression of Bcl-2 upon methionine deprivation may be the underlying mechanism to stimulate release of cytochrome c from mitochondria. PMID- 12527943 TI - A phase II protection study of BB-10010 in patients with high grade non-Hodgkin's lymphoma undergoing intensive chemotherapy. AB - The aim of this study was to determine whether administration of BB-10010, a synthetic stem cell inhibitor, would allow more intensive chemotherapy to be administered to patients with newly diagnosed high grade NHL. Thirteen patients were randomised to receive BB-10010 concurrently with dose-intensified BEMOP/CA chemotherapy (7 patients) or chemotherapy alone (6 patients). Although the mean neutrophil count of BB-10010 treated patients was higher following cycles 1, 2 and 3 of chemotherapy compared with those receiving chemotherapy alone, there was no difference in the mean number of cycles tolerated, blood component usage and hospital admissions due to infections. No specific toxicity of BB-10010 was identified. Whilst BB-10010 can be administered safely, it does not improve the ability of patients to tolerate intensive chemotherapy for high grade NHL. PMID- 12527944 TI - Epidermal growth factor receptor mRNA in peripheral blood of patients with pancreatic, lung, and colon carcinomas detected by RT-PCR. AB - The epidermal growth factor receptor (EGFR) is overexpressed in many solid tumors. An RT-PCR assay was used to detect EGFR mRNA in the blood of 43 patients with pancreatic, lung (NSCLC), colon, and renal carcinomas. After an initial serum tube was drawn to clear the line of detached epithelial cells, a 3 ml sample of EDTA-anticoagulated blood was collected from each cancer patient and healthy control. Total RNA was isolated from each sample and from cancer cell lines. After reverse transcription using specific priming, outer and nested primers for EGFR were employed for cDNA amplification. RNA integrity was confirmed with RT-PCR amplification using beta2-microglobulin primers. PCR products were electrophoresed on agarose gels containing ethidium bromide and visualized. The assay was validated using Southern blotting and was capable of detecting a lower limit of 100 fg of total RNA from the A431 cell line. EGFR positive cells were detected in 3/10 (30%) non-small cell lung cancer patients, 2/11 (18%) pancreatic cancer patients, and 2/16 (12.5%) colon cancer patients, but in 0/6 (0%) patients with renal carcinoma (16% of solid tumor patients overall). All 23 healthy controls were negative. This study is the first to apply RT-PCR for the detection of EGFR mRNA in the peripheral blood of patients with pancreatic and renal carcinomas, and it lends further support for the use of EGFR mRNA as a marker of CTCs in the blood of patients with certain types of solid tumors. PMID- 12527945 TI - Immunosensitization of resistant human tumor cells to cytotoxicity by tumor infiltrating lymphocytes. AB - Most anti-cancer therapies induce apoptotic cell death, but a major barrier to long-term cancer treatments is the generation of apoptosis-resistant tumor cells. Tumor cells that become resistant to one therapy are usually cross-resistant to subsequent therapies, including those with different cellular/molecular targets, suggesting that resistant tumor cells acquire modifications of the general apoptotic pathway. Most solid tumors are characterized by infiltration of lymphocytes (tumor infiltrating lymphocytes, TIL), which may serve as a basis for new strategies to generate tumor specific lymphocytes. However, TIL frequently are unable to kill autologous tumor cells suggesting that they are anergic/tolerant. It is possible that the TIL are functional but the tumor cells are resistant to TIL-mediated apoptotic pathways. Previous findings revealed that resistant tumor cells can be sensitized with cytokines or subtoxic concentrations of chemotherapeutic drugs and restore killing by cytotoxic lymphocytes. In this study, we examined whether TIL can kill autologous and allogeneic tumor cells following sensitization with chemotherapeutic drugs. Renal and prostate cancer derived TIL were cytotoxic to chemosensitized resistant tumor cells. Killing by TIL was found to be perforin-dependent and perforin-independent. These findings demonstrate that combination drug and immunotherapy may be able to overcome tumor cell resistance to killing by TIL. Further, in vivo sensitization of drug resistant tumor cells by subtoxic doses of sensitizing chemotherapeutic drugs may result in tumor regression by the host immune system. PMID- 12527946 TI - IL-11-induced reduction of C/EBP transcription factor binding may contribute to the IL-12 downregulation in tumor-bearing mice. AB - Using a mammary tumor model syngeneic to BALB/c mice, we have characterized several tumor-derived factors. We now report that the DA-3 cell line derived from this tumor, as well as the in vivo tumor itself, express IL-11. The expression of IL-11 in the tumor is detectable at the transcriptional and translational levels, as evidenced by RT-PCR and Western blots. Using a murine IL-11 ELISA, we observed no differences in IL-11 production between normal and tumor-bearer's macrophages or T cells, with or without activation. Interestingly, elevated levels of IL-11 were found in the sera of tumor-bearers, when compared to normal animals and even higher levels of IL-11 were detected in the tumor cystic fluid. Macrophages from mice bearing large mammary tumors show an impaired production of IL-12 and NO, whereas T cells from the same animals display a deficient production of IFN gamma. Pretreatment of normal macrophages with IL-11 resulted in no decrease in NO production, nor an impaired production of IFN-gamma was observed in normal T cells upon pretreatment with IL-11. However, pretreatment of normal macrophages with IL-11 resulted in a decreased production of IL-12, as revealed by ELISA and RT-PCR. Electromobility shift assays showed decreased binding of the transcription factor C/EBP to the IL-12p40 promoter of LPS-activated macrophages from normal animals, upon pretreatment with IL-11. In contrast, no differences were observed in the levels of NFkappaB binding under the same experimental conditions. Our results suggest that tumor-derived IL-11 may play a role in the depressed IL-12 production by macrophages, leading to the impaired immune functions observed during mammary tumorigenesis. PMID- 12527947 TI - Spontaneous mammary tumors in aging Noble rats. AB - Noble (Nb) rat strain has been used for the study of hormonal carcinogenesis of mammary and prostate glands, for its susceptibility to develop premalignant lesions as well as carcinomas in these organs by sex hormones. However, background information on the spontaneously developed mammary tumors in this rat strain is scarce. We report on the incidence rate, latency period and histopathology of mammary tumors spontaneously developed in the senile intact and untreated Nb rats compared with those induced by either combined treatments with sex steroids or 7,12-dimethylbenz[a]anthracene (DMBA) in the same rat strain. We observed that the incidence rate of spontaneous mammary tumors was 45% in female Nb rats and 3% in the males. The average age of the female Nb rats to develop palpable tumors was 14 months and rarely detected in animals less than 12 months old. It was also noted that the incidence rate of the spontaneous mammary tumors was similar to those induced by combined treatments with sex steroids for 8-10 months (46.7% for T+E2 and 55.6% for T+DES) but less than those by DMBA treatment in 8 months (over 80%). Histologically, majority of the spontaneous mammary tumors were fibroadenomas, which comprised 70% of all collected tumors and about 20% were carcinomas whereas tumors induced by steroid hormones and DMBA were all carcinomas. Distant metastases of spontaneous mammary carcinomas to lung, liver and lymph nodes were also noted, but rarely. PMID- 12527949 TI - Eating-related disorders in patients with advanced cancer. AB - Patients with advanced incurable illnesses and their families are frequently concerned about not eating enough and a decline in physical functioning. Eating related disorders were reviewed for a workshop at the 4th San Salvatore Meeting on Palliative Care in Switzerland, with the purpose of discussing new findings relating to the understanding and treatment of such problems. The topics discussed were patient and family concerns, practical nutritional advice, mechanism and causes of anorexia/cachexia, and assessment and management of eating-related disorders. PMID- 12527948 TI - Curative and palliative aspects of regional chemotherapy in combination with surgery. AB - Many attempts have been made in the last two decades to improve the outcome of patients with advanced or metastasised solid tumours. In particular, combined modality treatment strategies combining surgery with more localised therapies, e.g. radiotherapy, or systemic therapies such as chemotherapy have yielded promising data. The aim of regional chemotherapy is to improve locoregional cytostatic drug concentrations by achieving greater local efficacy and to diminish systemic side effects by reducing plasma drug levels. Highly qualified and experienced exponents of regional chemotherapy can complement surgical measures by applying multimodal strategies, because of their high efficacy in terms of tumour mass reduction without permanent tissue injuries, such as fibrosis or the damage to the vascular bed familiar from radiotherapeutic interventions. During the last 15 years, several new and very effective methods of administration, such as isolated pelvic perfusion or isolated thoracic perfusion, have extended the therapeutic arsenal of regional chemotherapy. The techniques needed for such transcutaneous and minimally invasive approaches as angiographically administered intra-arterial chemotherapy have been improved and side effects and the complication rate, dramatically reduced. Pharmacokinetic evaluations have demonstrated the high efficacy of one of the new regional modes of administration, isolated abdominal perfusion. With this technique, it is possible to attain cytostatic drug concentrations twice as high as those attained with systemic high-dose therapy with the same drug (treosulfan), but with only a quarter of the dosage and without bone marrow transplantation. Such techniques are now also available for the pelvic area, the thoracic region, the chest wall, the liver and the limbs. Regional chemotherapy is a very effective tool for induction therapy when tumours are apparently inoperable, as it can lead to sufficient shrinkage to make such tumours resectable. of all cases In an unselected series of 131 patients with colorectal liver metastases liver surgery with curative intent was possible after two cycles of therapy in 21.4% when immuno-chemoembolisation plus intra-arterial infusion was used as an inductive treatment. In 57.4% of these patients systemic chemotherapy had preceded surgery. After a median follow-up of more than 3 years, median survival has not been reached in the resected group and was 30 months for the group treated by regional chemotherapy alone. The aforementioned abdominal perfusion technique gave response rates in the range of 60-70% in patients with peritoneally metastasised and recurrent ovarian cancer. In about 55% of these cases, a second debulking procedure was possible, leading to a reduction in pain and symptoms and prolongation in survival, with a 1-year survival rate of 67%. Regional chemotherapy also seems to be very effective as an induction treatment in advanced non-small-cell lung cancer (NSCLC) patients in stages III A (bulky disease) and III B. After two cycles of isolated thoracic perfusion, the rate of remission was 86.3%. Removal of the remaining tumour structures was possible in 72.6% of all cases. Despite the high efficacy, the rate of side effects was low and acceptable. A steep increase in lung function parameters was observed in responding patients. This technique paves the way for a more effective induction therapy in advanced NSCLC, followed by resection and adjuvant radiotherapy of the mediastinum. PMID- 12527950 TI - Communication, information and support for adults with malignant cerebral glioma : a systematic literature review. AB - The goal of this work was a systematic review of communication, information and support for adults with malignant cerebral glioma. Medical, nursing and social science computerised databases up to spring 2000 were searched. Key organisations were contacted, and specialist journals were searched. Inclusion criteria were publication in English, inclusion of patients with malignant cerebral glioma, measurement of patients' and relatives' awareness of the prognosis, distress, satisfaction with information or care received, uptake of service or professional communication skills. Studies of patients with other cancers were excluded. Qualitative and quantitative studies were assessed, graded for methodological quality and combined. Twelve observational studies were found, although many were limited by sample selection, description and setting. Patient awareness of the prognosis varied, and relatives appeared more aware. There was no direct evidence about what patients and relatives wanted to know, but qualitative studies suggested that an individual approach to disclosure and maintaining hope were important. Most patients and relatives valued specialist nurse support highly. No specific studies of interventions to break bad news, giving information or training staff were found for these patients. Evidence from observational studies suggests that these patients need individually tailored communication and information, and specialist support. Existing intervention studies of patients with other cancers may suggest effective strategies. PMID- 12527951 TI - Clinical estimation of survival and impact of other prognostic factors on terminally ill cancer patients in Oman. AB - A prospective study was carried out in the only tertiary oncology department in Oman to analyse the pattern of various prognostic factors in terminally ill cancer patients and their impact on these patients. Between September 1999 and February 2001, terminally ill cancer patients with solid tumours who fulfilled the eligibility criteria were included. All of them were coded Do Not Resuscitate (DNR). Clinical estimation of survival as well as recording of different symptoms and signs was carried out for each patient. Survival was calculated from the date of the DNR coding to the date of death. A total of 162 patients were included. Mean survival time was 41.5 days (median 10 days). The gastrointestinal tract (GIT) was the most common site of malignant disease, followed by the breast. Univariate analysis of evaluable patients showed that performance status (PS), dry mouth, presence of delirium, anorexia, peripheral oedema, absence of bone metastasis, low lymphocyte count and low albumin level had significant effects on survival. Multiple regression analysis showed that PS and oedema were the only independent predictors of survival. Clinical prediction of survival was correlated with observed survival. Patient's PS, presence of peripheral oedema and clinical estimate of survival are good predictors of survival in terminally ill cancer patients. PMID- 12527952 TI - Pain and pain alleviation in hospital-based home care: demographic, biological and treatment factors. AB - The aim of this study was to contrast two opposed groups, namely palliative cancer patients who were suffering significant pain (VAS> or =4) and palliative cancer patients with no pain (VAS = 0) in hospital-based home care and, retrospectively, to study possible differences in relation to demographic, biological and treatment factors. The ESAS (Edmonton Symptom Assessment Scale) was used to assess 191 palliative cancer patients on admission and after 1 week of home care. Fifty-two (27%) had pain (mean 5.5+/-1.7) and 72 (38%) had no pain on admission [the middle group (n=67) had VAS 1-3]. Activity was more severely affected (5.4 vs 4.2, p<0.01) and nausea less well controlled in patients with pain (2.3 vs 0.7, P<0.0001). Pain was associated with the diagnosis of prostate cancer (P<0.01) and the presence of skeletal metastases (P<0.001), whereas pain free patients, with or without analgesics, more often had colorectal cancer (P<0.01) or melanoma (P<0.05). The medication profiles differed between the two groups: 22 (42%) of the 52 patients with pain were on step 3 of the WHO analgesic ladder and 24 of 51 (47%) were receiving antiemetics, whereas 42 (58%) of the 72 patients with no current pain had no analgesic prescribed and only 25% of them had antiemetics prescribed, indicating biological differences. If pain was present on admission a pain analysis was formally documented in 23 (44%) of the 52 cases and the medication was changed in 27 of the 52 (52%). The patients improved after 1 week (5.4+/-1.6 vs 3.9+/-2.3, P<0.001), and the improvement was significant even when a pain analysis was not documented or when medication was not changed. In conclusion, the results of this study indicate biological differences in pain alleviation and the need for a more structured way of working. PMID- 12527953 TI - Sucralfate mouthwash for prevention and treatment of 5-fluorouracil-induced mucositis: a randomized, placebo-controlled trial. AB - A randomized, double-blind, placebo-controlled trial was conducted to evaluate the effectiveness of a sucralfate mouthwash in preventing and alleviating oral mucositis induced by 5-fluorouracil (5FU). A total of 81 patients with colorectal cancer were enrolled. Patients were studied during their first cycle of chemotherapy with 5FU and leucovorin (LV) daily for 5 days every 4 weeks (Mayo Clinic schedule). Patients were randomly allocated to receive either a sucralfate suspension or a placebo suspension that was identical in appearance. Patients were instructed to use the suspension as a mouthwash four times daily from the beginning of the chemotherapy cycle. All patients received oral cryotherapy. Patients graded the severity of their own symptoms on a daily basis, and this was the primary outcome measure. There was no difference in the frequency or severity of oral mucositis between the sucralfate- and the placebo-treated group. Some mucositis was reported by 79% of the patient group. Assessment of mucositis by trial staff underestimated the incidence of this problem. Results of this trial do not support the hypothesis that a sucralfate mouthwash can prevent or alleviate oral mucositis induced by 5FU. Patient reporting of mucositis is a more sensitive instrument for assessment of mucositis than review by medical staff. PMID- 12527954 TI - Oral complications in Chinese cancer patients undergoing chemotherapy. AB - In this study the oral status and mucositis-related symptoms in 94 Chinese cancer patients undergoing chemotherapy for with curative intent were examined. Data were collected by a research nurse at three stages over a period of 16 days: at the start, on day 8, and on day 16 of chemotherapy. The majority of subjects experienced a mild to moderate degree of mucositis with the highest score reported on day 8. Patients with head and neck cancer had consistently higher scores for mucositis and symptoms in most phases. Oral mucositis and symptoms were significantly and positively correlated with consumption of spicy and/or hot (temperature) food, and frequency of use of mouthwashes. Significantly higher scores were found in patients who: ate hot (temperature) food, had low tolerance for hot food and spicy foods, and were taking Chinese medicinal herbs. Careful monitoring of patient diet preferences, cancer site, and use of Chinese medicine is recommended for comprehensive oral assessment. The alleviation of oral mucositis and the associated symptoms through promotion of an appropriate food/fluid intake and more frequent use of mouthwashes should be emphasized in oral care. PMID- 12527955 TI - Noninvasive ventilation: application to the cancer patient admitted in the intensive care unit. AB - The objective of this retrospective study of prospectively registered patients was to determine the usefulness and efficacy of noninvasive ventilation (NIV) in cancer patients admitted to the medical intensive care unit of an European cancer hospital for a medical complication, as reflected in discharges from the intensive care unit (ICU) and from hospital. The subjects were a total of 40 consecutive cancer patients (28 with solid tumours and 12 with haematological malignancies) who required immediate or delayed NIV. Variables relating to demographic parameters, severity scores, cancer characteristics, intensive care data and hospital discharge were recorded. The complications making NIV necessary were hypoxaemic pneumonia in 32.5%, hypercapnic ventilatory failure in 30%, multifactorial respiratory failure in 17.5%, acute haemodynamic oedema in 10%, acute respiratory distress syndrome in 2.5%, alveolar haemorrhage in 2.5%, pulmonary embolism in 2.5% and lysis pneumopathy in 2.5%. Most of the patients, 57.5% and 42.5%, respectively, were discharged from the ICU and from the hospital. Among the 10 patients (25%) who required salvage invasive mechanical ventilation, only 1 was discharged from hospital. Sixty-four per cent of the solid tumour patients and 42% of those with haematological malignancies were discharged from the ICU and 50% and 25%, respectively, from the hospital. NIV thus appears to be an effective form of ventilatory support for cancer patients, including those with solid tumours. PMID- 12527956 TI - Assessment of nutritional status and prognosis in advanced cancer: interleukin-6, C-reactive protein, and the prognostic and inflammatory nutritional index. AB - The Prognostic Inflammatory Nutritional Index (PINI) is a simple scoring system that has been used to evaluate nutritional status and prognosis in critically ill patients. The PINI has never been evaluated in advanced cancer. Fifty consecutive patients with advanced cancer, weight loss, and anorexia were studied. C-reactive protein (CRP), albumin, pre-albumin, interleukin-6 (IL-6), and alpha 1-acid glycoprotein (AAG) were evaluated. The individual values for AAG, CRP, and IL-6 were markedly elevated. In contrast to albumin and prealbumin, CRP levels were very high. The PINI was significantly elevated, and higher than reported in critically ill intensive care patients. Elevated IL-6 levels correlated with high PINI and CRP values. CRP, IL-6, and PINI should be considered in future research on nutritional status and prediction of prognosis in advanced cancer. PMID- 12527957 TI - Unrecognized thyroid dysfunction in patients with cancer. AB - Abnormal thyroid function can cause subtle medical symptoms in "healthy" individuals. Because cancer can cause similar symptoms, physicians may fail to suspect and test for thyroid abnormalities. We measured thyroid function in 158 patients with various types of cancers being followed at a comprehensive cancer center and found that 16% of them had a thyroid abnormality, which had not been diagnosed and treated in any of them. We conclude that thyroid dysfunction is usually unrecognized in cancer patients and may possibly contribute to their morbidity. PMID- 12527958 TI - Diazepam as an adjuvant analgesic to morphine for pain due to skeletal muscle spasm. AB - Side effects of morphine are common when it is given in titrated doses to control severe pain in advanced cancer. We describe a case of severe back pain resistant to parenteral morphine accompanied by muscle spasm, in which the addition of diazepam both had an opioid-sparing effect and provided superior symptomatic relief. Diazepam appears to have a specific role as an adjuvant analgesic for pain due to skeletal muscle spasm associated with painful vertebral metastases. PMID- 12527959 TI - Biomechanical and physiological aspects of legged locomotion in humans. AB - Walking and running, the two basic gaits used by man, are very complex movements. They can, however, be described using two simple models: an inverted pendulum and a spring. Muscles must contract at each step to move the body segments in the proper sequence but the work done is, in part, relieved by the interplay of mechanical energies, potential and kinetic in walking, and elastic in running. This explains why there is an optimal speed of walking (minimal metabolic cost of about 2 J.kg(-1).m(-1) at about 1.11 m.s(-1)) and why the cost of running is constant and independent of speed (about 4 J.kg(-1).m(-1)). Historically, the mechanical work of locomotion has been divided into external and internal work. The former is the work done to raise and accelerate the body centre of mass (m) within the environment, the latter is the work done to accelerate the body segments with respect to the centre of m. The total work has been calculated, somewhat arbitrarily, as the sum of the two. While the changes of potential and kinetic energies can be accurately measured, the contribution of the elastic energy cannot easily be assessed, nor can the true work performed by the muscles. Many factors can affect the work of locomotion--the gradient of the terrain, body size (height and body m), and gravity. The partitioning of positive and negative work and their different efficiencies explain why the most economical gradient is about -10% (1.1 J.kg(-1).m(-1) at 1.3 m.s(-1) for walking, and 3.1 J.kg(-1).m(-1) at between 3 and 4 m.s(-1) for running). The mechanics of walking of children, pigmies and dwarfs, in particular the recovery of energy at each step, is not different from that of taller (normal sized) individuals when the speed is expressed in dynamically equivalent terms (Froude number). An extra load, external or internal (obesity) affects internal and external work according to the distribution of the added m. Different gravitational environments determine the optimal speed of walking and the speed of transition from walking to running: at more than 1 g it is easier to walk than to run, and it is the opposite at less than 1 g. Passive aids, such as skis or skates, allow an increase in the speed of progression, but the mechanics of the locomotion cannot be simply described using the models for walking and running because step frequency, the proportion of step duration during which the foot is in contact with the ground, the position of the limbs, the force exerted on the ground and the time of its application are all different. PMID- 12527960 TI - Glycolysis and oxidative phosphorylation as a function of cytosolic phosphorylation state and power output of the muscle cell. AB - A mathematical description of the regulation of ATP production in muscle cells is presented whereby the activity of OxP can be calculated as a function of (1) free [ADP] as the substrate and (2) a second driving force PhiDelta G (kilojoules per mole) resulting from the difference of free energy Delta G(ox,ap) (kilojoules per mole)-Delta G(ATP,cyt) (kilojoules per mole). In turn, the term Delta G(ox,ap) results from the proton motive force and the generation of ATP in the matrix space including the ATP-ADP exchange, whereas the phosphorylation state of the CHEP-sytem is described by Delta G(ATP,cyt). Regulation of glycolysis is calculated as a function of free [ADP] and [AMP] at the level of PFK. The PFK is inhibited by a decreasing pH resulting from lactate accumulation. The ATP/PCr equilibrium of the CHEP-system is calculated by algebraic equations. The dynamic behaviour of the metabolic control of ATP production as a function of ATP consumption is calculated by a system of two 1st-order non-linear differential equations, including a time delay considering oxygen transport. Lactate distribution and elimination is calculated using a two-compartment model with an active lactate producing, and a passive, space including lactate elimination by combustion. The simulation of the dynamics of energy metabolism of muscle cells is performed by the stepwise solution of the differential equations with a 5th order Runge-Kutta-Fehlberg-routine. Examples of various applications of the simulation of the dynamics of energy supply demonstrate the qualitative and quantitative congruence to the behaviour of metabolic processes in experiments during rest, exercise and recovery. PMID- 12527961 TI - Parathyroid hormone concentrations during and after two periods of high intensity exercise with and without an intervening recovery period. AB - The purpose of this study was to examine the effect of a recovery period between two periods of exercise on bone metabolism and related hormones. Concentrations of serum parathyroid hormone ([PTH]), plasma ionized calcium ([Ca(2+)]) and total calcium were measured. A group of 12 healthy men aged 20-27 years participated in this study. They took part in two exercise protocols (P(1) and P(2)) on two separate weeks. The exercise in P(1) comprised two successive periods of 21 min each at 70% and 85% of maximal oxygen uptake; P(2) comprised two periods of exercise at the same intensities but separated by 40 min of recovery. Venous blood samples were collected 1 day before the sessions (control), before each protocol, during (7th and 21st min), at the end (42nd min in P(1) and 82nd min in P(2)) and after 24 h of recovery. The [PTH] was significantly elevated during the two protocols ( P<0.01), remained raised in P(1) after 24 h of recovery ( P<0.05) and was significantly lower ( P<0.01) at the end of P(2) when compared to P(1). The [Ca(2+)] decreased significantly during and at the end of the two protocols ( P<0.01) and had returned to control values after 24 h of recovery. Plasma lactate concentration increased during the two protocols ( P<0.01) and returned to control values after recovery. These results indicate firstly that [Ca(2+)] decreases during continuous exercise as [PTH] increases and remains raised after 24 h of recovery, secondly that a recovery period between two periods of exercise attenuates the variations in [Ca(2+)] and [PTH], and thirdly that recovery may have anabolic effects on bone. However, the small physiological changes observed prevent us from forming any firm conclusion about this. PMID- 12527962 TI - Effect of the menstrual cycle on performance of intermittent, high-intensity shuttle running in a hot environment. AB - The present study examined the impact of the menstrual cycle and oral contraceptive use on performance of high-intensity intermittent running in the heat [31.0 (0.2) degrees C; 23.1 (0.9)% relative humidity]. Seven normally menstruating women (NM) and eight oral contraceptive (OC) users participated in the study. Two trials were undertaken near the predicted mid-point of the follicular (FT) and luteal (LT) phases of the menstrual cycle and the equivalent days for the OC users. Basal serum progesterone concentrations were higher during the LT for the NM group [FT: 2.42 (0.28) nmol l(-1) vs LT: 25.96 (11.28) nmol l( 1); P<0.05], but were not different for the OC users [days 1-14: 2.79 (0.38) nmol l(-1) vs days 15-28: 2.61 (0.32) nmol l(-1)]. There were no differences in distance run between menstrual cycle phases or between the normally menstruating and OC groups [NM FT: 6257 (1401) m vs LT: 5861 (1035) m]. However, the OC ran further in days 15-28 compared to days 1-14 [OC 1-14: 5481 (612) m vs 15-28: 6615 (893) m, P<0.05]. For the NM, rectal temperature, perceived exertion, estimated SR, serum growth hormone, plasma lactate, ammonia and glucose did not differ between phases of the menstrual cycle. For the OC group, heart rate, perceived exertion, sweat rate, plasma lactate and ammonia did not differ between days 1-14 of OC use and days 15-28. However, rectal temperature was higher ( P<0.05) and growth hormone tended to be higher ( P=0.05) during days 15-28, while plasma glucose was lower ( P<0.05). These results demonstrate that for unacclimatised games players the performance of intermittent, high-intensity shuttle running in the heat is unaffected by menstrual cycle phase but is influenced by OC use. PMID- 12527963 TI - The 5 min running field test: test and retest reliability on trained men and women. AB - The repeatability of the maximal aerobic velocity ( v(amax)) estimated using the 5 min running field test (5(RFT)) has been examined in an heterogeneous population of 132 subjects distributed in five groups considering their sporting activities, their competition levels and their physical fitness levels: among them were national and local runners, rugby players, and multi-sport women and men. To establish the test and retest reliability, all the subjects took part in the 5(RFT) twice within 3 weeks. After the normality of distributions had been assessed using a Kolmogorov-Smirnov's test, a Student's paired t-test showed no difference between the two trials in all groups except that of the national runners. A heterogeneous group was then constituted from the other subjects, and this took part in the reliability study. Intraclass correlation coefficients calculated from a one-way ANOVA on the performances achieved by each group in both tests ranged from 0.94 to 0.98. The standard errors of measurement (SEM) of the 5(RFT) ranged from 0.95% (13 m) to 1.89% (20 m), which correspond to errors of 0.15 km.h(-1) and 0.34 km.h(-1) in the v(amax), respectively. These results indicate that the 5(RFT) is reliable when used in homogeneous groups with various characteristics as well as in a heterogeneous population. Moreover, the results of this study have shown that the 5(RFT) is reliable for estimating v(amax) from only one trial, since the intraclass correlation coefficients for one trial ranged from 0.88 to 0.96, which is of particular interest to coaches. Nevertheless, further studies would be necessary to evaluate the repeatability of this test in other populations such as school children and adults of both sexes having different characteristics. PMID- 12527964 TI - Maximal lactate steady state concentration (MLSS): experimental and modelling approaches. AB - It is assumed that the maximal lactate steady state (MLSS) can be used to establish the highest workload that can be maintained over time without continual blood lactate accumulation. In untrained subjects, and in both elite and junior athletes, MLSS occurs at different blood lactate concentrations (BLC) for different exercise modes. This suggests that MLSS depends on the motor pattern of exercise and may be a function of the relationship between power output per unit muscle mass and the mass of the muscle primarily engaged in the activity. A computer model has been developed that takes account of current theories relating to the effect of exercise on BLC and to the factors that limit oxygen transport to the muscle cell. Simulations using this model support the suggestion that load per unit of engaged muscle mass accounts for task-specific levels of MLSS. Simulated differences in MLSS appear because the MLSS does not necessarily reflect the real maximal equilibrium between lactate formation and utilization, the LLSS. The higher difference between MLSS and LLSS measured in rowing ergometry compared to cycle ergometry seems to indicate a greater task sensitivity of the BLC response to given changes of exercise intensity during rowing. Whether such a difference may be relevant for a deeper understanding of task-specific training strategies remains a matter for further investigation. PMID- 12527965 TI - Increased sensations of intensity of breathlessness impairs maintenance of intense intermittent exercise. AB - To identify the reserve of an individual's tolerance of the sensation of breathlessness and metabolic stress in maintaining intense intermittent exercise at exhaustion under conditions of normal breathing, the contribution of the effect of modest inspiratory load on these two responses to the change in the exercise sustainability (Ex(sus)) were examined. Seven men repeatedly performed 12 s exercise at 160% maximal aerobic power output followed by passive recovery for 18 s under normal and ventilatory muscle loaded (VML) breathing conditions until exhaustion. In the VML trial, ventilatory muscle work at exhaustion was double that of the normal control. The control Ex(sus) was reduced [mean (SEM)] [31.7 (6.6)%] while the slope of the time course for the rating of the perceived magnitude of breathing effort (RPMBE/Time), which reflected the intensity of breathlessness, was increased [164.8 (32.2)%] from control and the RPMBE at exhaustion was higher than corresponding control value [144.4 (21.8)%]. Moreover, increases in plasma ammonia and uric acid concentrations, which indicated metabolic stress, were increased [168.1 (28.0)% and 251.7 (57.4)%, respectively], with no change in total oxygen uptake from control when the control exercise was repeated with an identical duration of VML exercise. It was found that the reduction in Ex(sus) in the VML trial was correlated to the increase in their sensations of the intensity of breathlessness (RPMBE/Time: r=0.81; RPMBE at exhaustion: r=0.97, P<0.05). The reduction in Ex(sus), however, was not correlated to the increase in metabolite concentrations. These findings implied that there was no substantial reserve of tolerance of the sensation of breathlessness relative to that of metabolic stress in subjects maintaining intense intermittent exercise at exhaustion under normal conditions of breathing. PMID- 12527966 TI - Prolonged muscle vibration reduces maximal voluntary knee extension performance in both the ipsilateral and the contralateral limb in man. AB - Previous studies have shown that prolonged vibration of the rectus femoris decreases maximal voluntary knee extension performance in the ipsilateral leg. In the present study, measurements of maximal voluntary isometric knee extension contractions with the ipsilateral (right) leg and the contralateral (left) leg were made immediately before and after vibration treatment. Significant reductions in maximal force and maximum rate of force generation occurred in both the ipsilateral and contralateral legs following 30 minutes of continuous vibration at both 30 Hz and 120 Hz, with 30 Hz causing the greatest ipsilateral effects. However, although the level of neural activation (iEMG) of the vibrated muscle (right rectus femoris) was reduced following 30 Hz vibration ( P=0.026), there were no significant changes occurring in a synergistic muscle (right vastus lateralis) or in either contralateral muscle. It was concluded that muscle vibration may act through spinal reflex pathways to influence the homonymous motoneuron pool. The effects on contralateral force but not specific muscle iEMG suggest an effect on heteronymous motoneuron pools or an effect acting on central descending drive to contralateral muscles. These findings may have implications for the rehabilitation of patients with an immobilised limb. PMID- 12527967 TI - Maximal oxygen uptake during field running does not exceed that measured during treadmill exercise. AB - Modern ergometric equipment enables the simulation of laboratory maximal oxygen uptake (.VO(2max)) testing in the field. Therefore, it was investigated whether the improved event specificity on the track might lead to higher .VO(2max) measurements in running. Identical protocols were used on the treadmill and on the track (speed was indicated by a computer-driven flashing light system). Ambulatory measurements of gas exchange were carried out throughout both tests, which were executed in randomized order. There were no significant differences ( P=0.71) in .VO(2max) between treadmill [4.65 (0.51) ml.min(-1)] and field tests [4.63 (0.55) ml.min(-1)]. However, the test duration differed significantly ( P<0.001) by approximately 5%: treadmill 691 (39) s; field test 727 (42) s. With the exception of maximum heart rate (HR(max); significantly higher in the field with P=0.02) all criteria for the degree of effort were similar between the two tests. However, the difference in HR(max) at less than 2 beats.min(-1), was practically negligible. Submaximal measurements of oxygen uptake and minute ventilation were significantly higher on the treadmill ( P<0.001 for both parameters). In summary, field tests with incremental running protocols do not result in higher .VO(2max) measurements compared to laboratory treadmill exercise. A better running economy on the track results in higher maximal velocities and longer exercise durations being sustained. The determination of .VO(2max) is not a reasonable application for ambulatory gas exchange measurements because laboratory values are not surpassed. PMID- 12527968 TI - Changes in performance, maximal oxygen uptake and maximal accumulated oxygen deficit after 5, 10 and 15 days of live high:train low altitude exposure. AB - Nineteen well-trained cyclists (14 males and 5 females, mean initial .VO(2max) 62.3 ml kg(-1 )min(-1)) completed a multistage cycle ergometer test to determine maximal mean power output in 4 min (MMPO(4min)), maximal oxygen uptake (.VO(2max)) and maximal accumulated oxygen deficit (MAOD). The athletes were divided into three groups, each of which completed 5, 10 or 15 days of both a control condition (C) and live high:train low altitude exposure (LHTL). The C groups lived and trained at the ambient altitude of 610 m. The LHTL groups spent 8-10 h night(-1) in normobaric hypoxia at a simulated altitude of 2,650 m, and trained at the ambient altitude of 610 m. The changes to MMPO(4min), .VO(2max) and MAOD in response to LHTL altitude exposure were not significantly different for the 5-, 10- and 15-day treatment periods. For the pooled data from all three treatment periods, there were significant increases in MMPO(4min) [mean (SD) 5.15 (0.83) W kg(-1) vs 5.34 (0.78) W kg(-1)] and MAOD [50.1 (14.2) ml kg(-1) vs 54.9 (13.1) ml kg(-1)] in the LHTL athletes between pre- and post-altitude exposure. There were no significant changes in MMPO(4min) [5.09 (0.76) W kg(-1) vs 5.16 (0.86) W kg(-1)] or MAOD [50.5 (14.1) ml kg(-1) vs 49.1 (13.0) ml kg(-1)] in the C athletes over the corresponding period. There were significant increases in .VO(2max) in the athletes during both the LHTL [63.2 (9.0) ml kg(-1 )min(-1) vs 64.1 (9.0) ml kg(-1 )min(-1)] and C [62.0 (8.6) ml kg(-1 )min(-1) vs 63.4 (9.2) ml kg(-1 )min(-1)] conditions. In these athletes, there was no difference in the impact of 5, 10 or 15 days of LHTL on the increases observed in MMPO(4min), .VO(2max) or MAOD; and LHTL increased MMPO(4min) and MAOD more than training at low altitude alone. PMID- 12527969 TI - The effect of intermittent training in hypobaric hypoxia on sea-level exercise: a cross-over study in humans. AB - The purpose of this study was to examine the effect of intermittent training in a hypobaric chamber on physical exercise at sea level. Over a 10 day period, 16 male triathletes trained for 2 h each day on a cycle ergometer placed in a hypobaric chamber. Training intensity was at 60%-70% of the heart rate reserve. There were 8 subjects who trained at a simulated altitude of 2,500 m, the other 8 trained at sea level. A year later, a cross-over study took place. Baseline measurements were made on a cycle ergometer at sea level, which included an incremental test until exhaustion and a Wingate Anaerobic Test. Altogether, 12 subjects completed the cross-over study. At 9 days after training in hypoxia, significant increases were seen in maximal power output (.W(max))(5.2%), anaerobic mean power (4.1%), and anaerobic peak power (3.8%). A non-significant increase in maximal oxygen uptake (.VO(2max)) of 1.9% was observed. At 9 days after training at sea level, no significant changes were seen in .W(max)(2.1%), .VO(2max) (2.0%), anaerobic mean power (0.2%) and anaerobic peak power (0.2%). When comparing the results of the two training regimes, the anaerobic mean power was the only variable that showed a significantly larger increase as a result of training at altitude. And, although the differences in percentage change between the two training protocols were not significant, they were substantial for as well as for anaerobic peak power. The results of this study indicate that intermittent hypobaric training can improve the anaerobic energy supplying system, and also, to a lesser extent, the aerobic system. It can be concluded that the overall results of the cross-over study showed predominantly improvements in the anaerobic metabolism at variance with the previous study of our own group, where the relative .VO(2max) and .W(max) increased by 7%. PMID- 12527970 TI - Effect of magnitopuncture on sympathetic and parasympathetic nerve activities in healthy drivers--assessment by power spectrum analysis of heart rate variability. AB - The object of this study was to assess the effects of magnitopuncture applied to Dazhui (DU14) point and Neiguan (PC6) points on sympathetic and parasympathetic nerve activities by power spectrum analysis of heart rate variability in healthy drivers during simulated driving. Using power spectrum analysis, the low frequency (LF) and high frequency (HF) components of heart rate variability can be calculated reflecting the sympathetic and parasympathetic activity. The 40 healthy male subjects were randomly divided into two groups: A (study group) and B (control group). All subjects were required to be well rested before the experiment. The subjects of both groups were required to perform a simulated driving task for 3 h. During the driving, magnitopunctures were applied to the DU14 and PC6 points for A while the subject performed the task for 2.5 h, and for B magnitopunctures were applied to non-acupuncture points which were 1.5 cm away from the two acupuncture points respectively over the same time. Subjective response to a questionnaire was obtained after the simulated task in the two groups. At the end of the driving task the LF component in normalized units (NU) had decreased significantly ( P<0.05) indicating a reduced sympathetic nerve activity and the HF component (NU) increased significantly ( P<0.05) indicating a increased parasympathetic nerve activity for A compared with pre-stimulation while for B no significant differences were observed. There were significant group differences in LF (NU), HF (NU) and LF:HF at the end of the driving task ( P<0.05). It was concluded that a modulating effect of magnitopuncture on sympathetic and parasympathetic nerve activities in healthy subjects was associated with the acupuncture points. The findings represent physiological evidence that magnitopuncture may reduce mental fatigue in healthy drivers. PMID- 12527971 TI - A comparison of two time-domain analysis procedures in the determination of .VO(2) kinetics by pseudorandom binary sequence exercise testing. AB - The purpose of this study was to apply and compare two time-domain analysis procedures in the determination of oxygen uptake (.VO(2)) kinetics in response to a pseudorandom binary sequence (PRBS) exercise test. PRBS exercise tests have typically been analysed in the frequency domain. However, the complex interpretation of frequency responses may have limited the application of this procedure in both sporting and clinical contexts, where a single time measurement would facilitate subject comparison. The relative potential of both a mean response time (MRT) and a peak cross-correlation time (PCCT) was investigated. This study was divided into two parts: a test-retest reliability study (part A), in which 10 healthy male subjects completed two identical PRBS exercise tests, and a comparison of the .VO(2) kinetics of 12 elite endurance runners (ER) and 12 elite sprinters (SR; part B). In part A, 95% limits of agreement were calculated for comparison between MRT and PCCT. The results of part A showed no significant difference between test and retest as assessed by MRT [mean (SD) 42.2 (4.2) s and 43.8 (6.9) s] or by PCCT [21.8 (3.7) s and 22.7 (4.5) s]. Measurement error (%) was lower for MRT in comparison with PCCT (16% and 25%, respectively). In part B of the study, the .VO(2) kinetics of ER were significantly faster than those of SR, as assessed by MRT [33.4 (3.4) s and 39.9 (7.1) s, respectively; P<0.01] and PCCT [20.9 (3.8) s and 24.8 (4.5) s; P<0.05]. It is possible that either analysis procedure could provide a single test measurement of .VO(2) kinetics; however, the greater reliability of the MRT data suggests that this method has more potential for development in the assessment of .VO(2) kinetics by PRBS exercise testing. PMID- 12527972 TI - Role of nitric oxide in isometric contraction properties of rat diaphragm during hypoxia. AB - Hypoxia disturbs Ca(2+) regulation and increases the intracellular Ca(2+) concentration ([Ca(2+)](i)), which may in turn activate the nitric oxide synthase (NOS) regulated by [Ca(2+)](i). Since nitric oxide (NO) reduces the isometric contractility of rat diaphragm in vitro, we hypothesized that NO contributes to the impaired force generation of an hypoxic diaphragm. The effects of different concentrations of the NOS inhibitor, N(G)-monomethyl-L-arginine (L-NMMA), the NO scavenger haemoglobin (150 micro mol.l(-1)) and the NO donor spermine NONOate (Sp NO; 1 mmol.l(-1)) were determined on isometric contractility during hypoxia [partial pressure of oxygen, PO(2), about 7 kPa (about 54 mmHg)] and hyperoxia [ PO(2) about 83 kPa (about 639 mmHg)]. Hypoxia significantly reduced maximal twitch force ( F(t)), and submaximal tetanic force (30 Hz, F(30)) in all L-NMMA groups. A low concentration of L-NMMA (30 micromol.l(-1)) increased F(30) but a high concentration (1,000 micromol.l(-1)) reduced F(30) during hypoxia. The effects of L-NMMA on force generation were more pronounced during hypoxia compared to hyperoxia. Peak increases in F(30) and F(t) were observed at a concentration of 30 micromol.l(-1) L-NMMA during hypoxia, but with 10 micromol.l( 1) L-NMMA during hyperoxia. The same concentration of haemoglobin increased F(30) and F(t) less during hypoxia compared to hyperoxia. The Sp-NO reduced F(t), F(30) and maximal tetanic force (F(0)) during hypoxia; these effects were abolished in the presence of haemoglobin. The Sp-NO did not alter F(t), F(30) and F(0)during hyperoxia. We conclude that NO plays a more prominent role during hypoxia and that NO contributes to the depression of force generation in the hypoxic rat diaphragm in vitro. This change may be related to an elevated NO generation within the hypoxic diaphragm. PMID- 12527973 TI - Physiological characteristics of elite short- and long-distance triathletes. AB - The purpose of this study was to compare the physiological responses in cycling and running of elite short-distance (ShD) and long-distance (LD) triathletes. Fifteen elite male triathletes participating in the World Championships were divided into two groups (ShD and LD) and performed a laboratory trial that comprised submaximal treadmill running, maximal then submaximal ergometry cycling and then an additional submaximal run. "In situ" best ShD triathlon performances were also analysed for each athlete. ShD demonstrated a significantly faster swim time than LD whereas .VO(2max) (ml kg(-1) min(-1)), cycling economy (W l(-1) min( 1)), peak power output (.W(peak),W) and ventilatory threshold (%.VO(2max)) were all similar between ShD and LD. Moreover, there were no differences between the two groups in the change (%) in running economy from the first to the second running bout. Swimming time was correlated to .W(peak)(r=-0.76; P<0.05) and economy ( r=-0.89; P<0.01) in the ShD athletes. Also, cycling time in the triathlon was correlated to .W(peak)(r=-0.83; P<0.05) in LD. In conclusion, ShD triathletes had a faster swimming time but did not exhibit different maximal or submaximal physiological characteristics measured in cycling and running than LD triathletes. PMID- 12527974 TI - Metabolic profile of 4 h cycling in the field with varying amounts of carbohydrate supply. AB - Several laboratory studies have demonstrated a performance-enhancing effect of carbohydrate (CHO) supplementations during endurance sessions of long duration. However, the transferability of these results to real training and competition circumstances has not been conclusively shown. Therefore, we tried to test the influence of graded CHO substitution on substrate utilization and selected physiological parameters under standardized but practically orientated field conditions. Fourteen endurance-trained male subjects [mean (SD): 25 (5) years, 72 (9) kg, .VO(2max) 67 (6) ml.min(-1).kg(-1), individual anaerobic threshold (IAT) 269 (30) W] after a stepwise increasing pre-test had to perform three 4-h endurance rides on their own bicycles with simultaneous spiroergometry: constant workload 70% IAT (monitoring by SRM-System). Before and during exercise, solutions without (0%), with 6% or 12% CHO were administered double-blindly and in randomized order (total volume: 50 ml.kg(-1)). After cessation of exercise, significant differences between 0% and both CHO concentrations were detected for blood glucose (GLU; 75 mg dl(-1) for 0% vs 101 mg dl(-1) for 6% vs 115 mg dl(-1) for 12%; P<0.001) and respiratory exchange ratio (0.84 vs 0.88 vs 0.90; P<0.01; correlation to GLU: r=0.46, P<0.05). Free fatty acids (0.19 vs 0.16 vs 0.10 mmol l(-1)) and glycerol (0.41 vs 0.22 vs 0.12 mmol l(-1)) were significantly different between the endurance trials in a dose-dependent manner (both P<0.001). Lactate concentration ( P=0.42) and heart rate ( P=0.12) had no significant influence from CHO substitution. We conclude that CHO substitution during 4-h endurance training inhibits lipolysis in a dose-dependent manner and enhances aerobic glycolysis. This proves that earlier laboratory findings can be replicated under field conditions using modern portable equipment. PMID- 12527975 TI - The effect of even, positive and negative pacing on metabolic, kinematic and temporal variables during breaststroke swimming. AB - The aim of this investigation was to determine the effect of even, negative and positive pacing on metabolic, kinematic and temporal variables during breaststroke swimming. Nine male swimmers [mean (SD): age 21 (3) years, height 178 (5) cm, body mass 77.2 (6.7) kg, 200 m-time 158.6 (13.6) s] completed a 200-m breaststroke time trial and then 72 h later three paced (even, positive, negative) 175-m breaststroke swims in random order, 48 h apart. The swimmers paced accurately in all trials. The evenly paced trial produced lower post exercise peak blood lactate and rating of perceived exertion values compared to the positively paced trial ( P<0.05). Peak oxygen uptake was not significantly different between trials ( P>0.05). Heart rate immediately following exercise was lower in the negatively paced trial ( P<0.05) than the other paced trials. Stroke rates were lower during the first half of the evenly and negatively paced trials compared to the positively paced trial ( P<0.01), but no differences were observed between the second half of the trials. Across all trials the stroke count increased as the trials progressed ( P<0.01). Turning times were observed to be shorter during the first half of the positively paced trial compared to the other paced trials ( P<0.01). Even paced swimming appears to be less physically stressful than positively paced swimming during high intensity exercise, as indicated by a lower post-exercise blood lactate concentration, perceived exertion and variability within turning times. PMID- 12527976 TI - Effects of pre-exercise ingestion of differing amounts of carbohydrate on subsequent metabolism and cycling performance. AB - Studies on the effect of the pre-exercise ingestion of carbohydrate on metabolism and performance have produced conflicting results, perhaps because of differences in the designs of the studies. The purpose of the present study was to examine the effects of ingesting differing amounts of glucose pre-exercise on the glucose and insulin responses during exercise and on time-trial (TT) performance. Nine well-trained male cyclists completed four exercise trials separated by at least 3 days. At 45 min before the start of exercise subjects consumed 500 ml of a beverage containing either 0 g (PLAC), 25 g (LOW), 75 g (MED) or 200 g (HIGH) of glucose. The exercise trials consisted of 20 min of submaximal steady-state exercise (SS) at 65% of maximal power output immediately followed by a [mean (SEM)] 691 (12) kJ TT. Plasma insulin concentrations at the onset of exercise were significantly higher ( P<0.05) in MED and HIGH compared with LOW and PLAC. Plasma glucose concentration fell rapidly ( P<0.05) during SS exercise in all glucose trials, but remained steady in PLAC. No difference in plasma glucose concentration was observed between the glucose trials at any time. Hypoglycaemia (less than 3.5 mmol.l(-1)) was observed in six subjects during SS but only after ingesting glucose pre-exercise. However, there was no difference in TT performance between the four trials. The ingestion of 0, 25, 75 or 200 g of glucose 45 min before a 20 min submaximal exercise bout did not affect subsequent TT performance. In addition, mild rebound hypoglycaemia following pre-exercise glucose ingestion did not negatively affect performance. PMID- 12527977 TI - Effects of timing of pre-exercise ingestion of carbohydrate on subsequent metabolism and cycling performance. AB - The occurrence of rebound hypoglycaemia may depend on the timing of carbohydrate intake. The aim of the present study was to investigate the metabolic and performance responses to the ingestion of carbohydrate at differing times before exercise. Eight subjects [mean (SEM)] [28 (3) years, 74.5 (2.6) kg, maximal oxygen uptake 63.1 (3.1) ml.kg(-1).min(-1)] performed three experiments. They ingested 75 g glucose dissolved in 500 ml water, thereafter resting for either 15, 45 or 75 min (15-Pre, 45-Pre and 75-Pre) before exercising for 20 min at 65% maximal power output followed by a time trial [total work 685 (18) kJ]. There were no differences in performance between conditions [mean powers 268 (10), 269 (7) and 276 (12) W for 15-Pre, 45-Pre and 75-Pre, respectively]. There were significant differences in plasma glucose concentration between 15-Pre [6.6 (0.6) mmol.l(-1); P<0.05] and both 45-Pre [4.5 (0.2) mmol.l(-1)] and 75-Pre [3.7 (0.2) mmol.l(-1)] immediately before exercise. Insulin concentrations immediately before exercise were higher ( P<0.05) during 15-Pre [72.6 (10.4) microU.ml(-1)] than during 45-Pre [50.8 (9.9) microU.ml(-1)], which was higher ( P<0.05) than during 75-Pre [33.9 (5.5) microU.ml(-1)]. These differences disappeared within 10 min of exercise. Two subjects became hypoglycaemic (plasma glucose concentration of less than 3.5 mmol.l(-1)) in the 15-Pre while three and five subjects were transiently hypoglycaemic in the 45-Pre and 75-Pre, respectively. Performance and rating of perceived exertion did not seem to be related to hypoglycaemia. Altering the timing of the ingestion of carbohydrate before exercise resulted in differences in plasma glucose/insulin responses which disappeared within 10 min of exercise and which had no effect on performance. Hypoglycaemia was observed in some subjects during the first 10 min but this did not affect performance. PMID- 12527978 TI - Effects of pre-exercise ingestion of trehalose, galactose and glucose on subsequent metabolism and cycling performance. AB - The glycaemic and insulinaemic responses to different carbohydrates vary and these have been suggested to affect performance. The purpose of the present study was to determine the effects of pre-exercise ingestion of glucose (GLU), galactose (GAL) and trehalose (TRE) on metabolic responses at rest and during exercise and on subsequent time-trial (TT) performance. Eight well-trained male cyclists completed three exercise trials separated by at least 3 days. At 45 min before the start of exercise subjects consumed 500 ml of a beverage containing 75 g of either glucose, galactose or trehalose. The exercise trials consisted of 20 min of submaximal steady-state exercise (SS) at 65% of maximal power output immediately followed by a [mean (SEM)] 702 (25) kJ TT. Plasma glucose concentration 15 min postprandial was significantly higher in GLU compared to GAL and TRE ( P<0.05). This was accompanied by a more than twofold greater rise in plasma insulin concentration in GLU compared to GAL and TRE (118% and 145%, respectively). During SS exercise four subjects in GLU and one subject in TRE developed a rebound hypoglycaemia (plasma glucose concentration less than 3.5 mmol.l(-1)). No differences were observed in TT performance between the three trials. Pre-exercise ingestion of trehalose and galactose resulted in lower plasma glucose and insulin responses prior to exercise and reduced the prevalence of rebound hypoglycaemia. Despite the attenuated insulin and glucose responses at rest and during exercise following pre-exercise ingestion of galactose and trehalose, there was no difference in TT performance compared with pre-exercise ingestion of glucose. PMID- 12527979 TI - Effects of pre-exercise ingestion of carbohydrate on glycaemic and insulinaemic responses during subsequent exercise at differing intensities. AB - The development of rebound hypoglycaemia has been reported after pre-exercise carbohydrate (CHO) ingestion in some studies but not in others. Differences in the experimental design and factors such as the exercise intensity are likely to be responsible for the discrepancies between these studies. Exercise intensity might be a crucial factor since it affects both insulinaemia and glucose uptake. Therefore the aim of the present study was to compare the glycaemic and insulinaemic responses to exercise at different intensities after ingestion of a standardized pre-exercise CHO load. Eight moderately trained subjects consumed 75 g of glucose 45 min prior to 20 min of exercise at 40%, 65% or 80% maximal power output. Blood samples were collected before glucose ingestion, at 15 min intervals at rest and 5 min intervals during exercise. During exercise, measurements of heart rate and breath-by-breath analysis of expired gas were performed continuously. The trials were performed at [mean (SEM)] 55 (1), 77 (1) and 90 (1) percentages maximal oxygen uptake. At the onset of exercise, plasma glucose concentration returned to pre-ingestion levels, while the insulin concentration was more than three times higher than at rest [on average 57 (7) compared to 16 (1) microU.ml(-1)]. During exercise, plasma glucose concentrations decreased during the first 5 min of exercise and then stabilized in all trials at concentrations that would not be considered to be hypoglycaemic. There were no significant differences in glucose or insulin concentrations between the three trials during exercise. These data suggest that the glycaemic response to ingestion of 75 g of CHO 45 min pre-exercise is similar during exercise of different intensities. PMID- 12527980 TI - Short-term effects of whole-body vibration on maximal voluntary isometric knee extensor force and rate of force rise. AB - Whole-Body vibration (WBV) may lead to muscle contractions via reflex activation of the primary muscle spindle (Ia) fibres. WBV has been reported to increase muscle power in the short term by improved muscle activation. The present study set out to investigate the acute effects of a standard WBV training session on voluntary activation during maximal isometric force production (MVC) and maximal rate of force rise (MRFR) of the knee extensors. Twelve students underwent a single standard WBV training session: 5x1 min vibration (frequency 30 Hz, amplitude 8 mm) with 2 min rest in between. During vibration, subjects stood barefoot on the vibration platform with their knees at an angle of 110 degrees. At 90 s following vibration, maximal voluntary knee extensor force was reduced to 93 (5)% [mean (SD), P<0.05] of baseline value and recovered within the next 3 h. Voluntary activation remained significantly depressed (2-4%). Neither the electrically induced MRFR nor voluntary MRFR were significantly affected by WBV. In addition, six WBV training sessions in 2 weeks ( n=10) did not enhance either voluntary muscle activation during MVC [99 (2)% of the baseline value] or voluntary MRFR [98 (9)% of the baseline value]. It is concluded that in the short term, WBV training does not improve muscle activation during maximal isometric knee extensor force production and maximal rate of force rise in healthy untrained students. PMID- 12527981 TI - Influence of cycling cadence on neuromuscular activity of the knee extensors in humans. AB - The aim of the present study was to investigate the influence of pedalling rate and power output in cycling on the neuromuscular activity of the knee extensor muscles. Ten subjects took part in 15 randomised trials, which consisted of three levels of power outputs (60%, 80% and 100% maximal aerobic power) and five cadences (70%, 85%, 100%, 115% and 130% of the freely chosen cadence, FCC). Root mean square (rms) was utilized to quantify electromyographic activity of the vastus lateralis (VL), vastus medialis (VM) and rectus femoris (RF) muscles. The mean (SD) FCC did not change with power output, ranging from 85.0 (11.9) to 88.0 (11.1) rpm. A significant power effect ( P<0.01) for the rms of VL, VM and RF muscles was observed. Results showed no significant cadence effect on neuromuscular activity of the VL and VM muscles, while the rms of the RF muscle was significantly greater ( P<0.05) at 70% FCC when compared to other cadences. In conclusion, the neuromuscular activity of the knee extensor muscles was not significantly influenced by cadence manipulations. Thus, minimisation of the neuromuscular activity of these muscles would not seem to lead to the choice of a cadence in cycling. PMID- 12527982 TI - Resting thyroid and leptin hormone changes in women following intense, prolonged exercise training. AB - This study examined whether free (f) triidothyronine (T3), f thyroxine (T4), thyroid stimulating hormone (TSH), and leptin concentrations at rest changed in response to 20 weeks of exercise-training. Two groups of women were recruited for participation in the study, collegiate athletes ( n=17) and sedentary controls (n=4). Exercise training consisted of daily athletic activity such as rowing, running, and weight lifting. Subjects were initially grouped into rowers and controls. However, earlier suggested criteria were further used to categorize hormone changes (percentages) in the subjects into (+) responders (increases), ( ) responders (decreases), or non-responders (no changes). The fT3 results of the rowers revealed two distinct categories of responses, (-) responder (all decreases; n=10) and non-responder (no change; n=7) rowers. In the responders fT3 concentration decreased (P<0.05) from baseline (BL) during an intense training period [(mean SEM) at 5 weeks by -28.2 (6.2)% and at 10 weeks by -24.9 (7.9)%], then returned towards BL levels (20 weeks compared to BL, P>0.05). Similar changes (P<0.05), at comparable times, were noted for leptin and TSH concentrations in the (-) responder rowers. The non-responder rowers and control subjects displayed no significant (P>0.05) hormone changes over the 20 weeks. The hormone changes observed in the (-) responder rowers were not significantly (P>0.05) correlated with changes in body composition or hydration status during the study. The mechanism for the hormone changes in the (-) responder rowers is unclear. We speculate the decrease in concentrations of TSH and fT3 could be attributable to a lower hypothalamic-pituitary signaling action, and this is related to the decreased leptin concentrations, and could represent a possible means of energy conservation in these exercising women. PMID- 12527983 TI - Position sense acuity is diminished following repetitive low-intensity work to fatigue in a simulated occupational setting: a critical comment. PMID- 12527984 TI - Frontal syndrome and disorders of executive functions. AB - The study of executive functions began with the early description of behavioural disorders induced by frontal damage. The development of experimental neuropsychology has led to the description of a large variety of cognitive disorders. Such approach has generated numerous tests that are used in clinical practice. More recently, theoretical approaches have proposed an organisation of executive processes and have documented the diversity of executive functions and related anatomy. These studies have deeply influenced the clinical approach, the assessment and the diagnosis of planning and executive disorders. For clinical practice, these data have favoured specific assessment of some key behavioural and cognitive deficits based on a battery of tests. PMID- 12527985 TI - Chorea Sancti Viti in Lexicon medicum anno 1696. PMID- 12527986 TI - Influence of prestroke dementia on early and delayed mortality in stroke patients. AB - Causes of early and delayed death after stroke differ. It has been suggested that delayed mortality rate was increased in patients with post-stroke dementia. Prestroke dementia is frequent: its influence on survival in stroke patients has never been evaluated. The aim of this study was to evaluate the influence of prestroke dementia on early and delayed mortality rate after stroke. In a cohort of 202 consecutive stroke patients aged >or= 40 years admitted between November 1995 and May 1996 in a primary care center, the prevalence of prestroke dementia was determined using the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE) with a cut-off of 104. Patients were followed-up for 3 years. Statistics were performed using life-table methods. Of 202 patients, 33 had prestroke dementia. Of 142 survivors at month-6, 44 were demented, of them 15 having prestroke and 29 new-onset post-stroke dementia. No patient was lost to follow-up. The risk of death at month-6 was higher in patients with prestroke dementia (RR 2.7; 95 % CI: 1.6-4.8). However, independent predictors of early death were age, severity of the deficit at admission, type and etiology of stroke. The risk of delayed death was higher in patients with prestroke dementia (RR 4.97; 95 % CI: 1.76-13.98) as in patients with new-onset post-stroke dementia (RR 6.24; 95 % CI: 2.67-14.57), compared with non-demented patients. The mortality rate did not differ between patients with prestroke and new-onset post stroke dementia. Dementia at month-6 was an independent predictor of delayed death (RR 5.7; 95 % CI: 2.4-13.4), with age and stroke recurrence. Causes of death did not differ between demented and non-demented patients. Dementia adversely influences vital outcome in stroke patients, perhaps partly because the therapeutic approach differs between demented and non-demented patients. PMID- 12527987 TI - GM2 gangliosidosis variant B1 neuroradiological findings. AB - Variant B1 is a rare type of GM2 gangliosidosis. Clinically, it shows a wide spectrum of forms ranging from infantile to juvenile. We report the first magnetic resonance imaging (MRI) findings from three patients affected by GM2 gangliosidosis variant B1, two presenting with the infantile form and one with the juvenile form. The MRI appearances of the two patients with the infantile form disease are congruent with those reported for the early-onset type of both Tay-Sachs and Sandhoff diseases, and are characterized by early involvement of the basal ganglia and thalamus with cortical atrophy appearing later. In contrast, the patient with the juvenile form of variant B1 showed progressive cortical and white-matter atrophy of the supratentorial structures and, to a lesser extent, the infratentorial structures. No basal ganglia or thalamic anomalies were observed. Because in the adult forms of both Tay-Sachs and Sandhoff diseases a progressive cerebellar atrophy represents the only abnormality detectable, it appears that an MRI pattern peculiar to GM2 gangliosidosis can be defined. This pattern ranges from the basal ganglia injury associated with the early and severe demyelination process noted in the infantile form of the disease, to cerebellar atrophy with no supratentorial anomalies in the adult form. An "intermediate" MRI picture, with cortical atrophy and mild cerebellar atrophy, but without basal ganglia impairment, can be observed in the juvenile form. In addition, our investigations suggest that MRI abnormalities in GM2 gangliosidosis correlate with the clinical form of the disease rather than with the biochemical variant of the enzymatic defect. PMID- 12527988 TI - Evidence of Wallerian degeneration in normal appearing white matter in the early stages of relapsing-remitting multiple sclerosis: a HMRS study. AB - OBJECTIVE: Wallerian degeneration in normal appearing white matter in early relapsing-remitting multiple sclerosis (RRMS), and its correlation with the number of relapses and disease duration. Background Recent pathological studies have demonstrated Wallerian degeneration in normal appearing white matter (NAWM) in multiple sclerosis (MS), in established RRMS, and in chronic MS. However, the presence of Wallerian degeneration early in the disease and its correlation with relapse and with disease duration has not been studied. METHODS: We performed proton magnetic resonance spectroscopic imaging in 21 MS patients, and 4 healthy controls, age and gender matched, aged under 45 years, with a maximum of 4 years since first bout, and an EDSS score of less than 3.0. N-acetyl-aspartate (NAA) (an index of axonal integrity) was measured in the NAWM from the pons and the cerebellar peduncles. RESULTS: We observed that the NAA levels were abnormally low in the NAWM in the early RRMS patients (p = 0.04, Student's t-test). The decrease in the NAA concentration correlated with disease duration in the two areas studied (p = 0.03 for pons and p = 0.04 for cerebellar peduncle); and with the number of previous relapses (Pearson's correlation = -0.582, p < 0.002). CONCLUSION: Wallerian degeneration measured by the NAA concentration at pons and cerebellar peduncles is present early in the disease and correlates with the number of relapses and disease duration. PMID- 12527989 TI - Cerebral venous thrombosis 3-year clinical outcome in 55 consecutive patients. AB - An early diagnosis and heparin therapy have contributed to a decreased mortality in cerebral venous thrombosis (CVT). However, predictors of outcome are difficult to identify, because most studies suffered heterogeneity in diagnostic findings and treatments, retrospective design, and recruitment bias. The aim of this study was to evaluate the clinical outcome in 55 consecutive patients with CVT admitted over a 4-year period. The study population consisted of 42 women and 13 men, with a median age of 39 years (range 16-68). The diagnosis was performed with MRI in 53 patients, and angiography in 2. The outcome was assessed with the modified Rankin scale (mRs). After a median follow-up of 36 months (range: 12-60), 45 patients were independent (mRS 0-2), and 10 were dependent or dead (mRS 3-6). Of 48 survivors, 7 had seizures, 6 motor deficits, 5 visual field defects, 29 headache (migraine in 14, tension headache in 13, other in 2). The logistic regression analysis found focal deficits and cancer at time of diagnosis, as independent predictors of dependence or death at year 3, and isolated intra cranial hypertension as an independent predictor of survival and independence. Mortality rates are low in the absence of cancer and focal deficits, and more than 80 % of survivors are independent after 3 years. However, 3/4 of survivors have residual symptoms. Therefore, despite a low mortality rate, CVT remains a serious disorder. PMID- 12527990 TI - Verbal laterality and handedness in patients with intracranial arachnoid cysts. AB - Left-handedness is most often genetic, but may also follow early, localised damage to the developing brain. This so-called "pathological" left-handedness syndrome is often associated with right hemisphere speech dominance. To find out whether verbal laterality or handedness were affected by congenital, intracranial arachnoid cysts, 51 consecutive patients with temporal or frontal arachnoid cysts were tested for handedness and verbal laterality, as measured with the dichotic listening (DL) technique. Handedness was normal in all subgroups of patients. In the preoperative DL test, only 51 % of the patients showed the normal superiority of the right ear (Right Ear Advantage - REA), significantly different from the REA frequency (74 %) in a normal reference group. Patients with a left temporal, or a frontal cyst had significantly lower preoperative REA frequencies than the reference group, whereas patients with a right temporal cyst did not differ from the reference group. Three to six months after decompressive surgery, the REA frequency (73 %) in the cyst patients was no longer different from that of the reference group. This postoperative normalization was seen both in patients with a left temporal or a frontal cyst. It is concluded that arachnoid cysts may suppress cognitive, cortical functions, and that this suppression can be reversed by surgical decompression of the cyst, even in adults. In our opinion, this cognitive "normalization" may in itself be an indication for decompressive surgery. It is further concluded that, although such cysts are congenital, the pressure from the cyst on the adjacent brain is not strong, nor persistent enough to cause a pathological left-handedness. PMID- 12527991 TI - Abnormalities of cortical excitability and cortical inhibition in cervical dystonia Evidence from somatosensory evoked potentials and paired transcranial magnetic stimulation recordings. AB - Cortical excitability and cortico-cortical inhibition were examined in twenty-one patients suffering from idiopathic rotational cervical dystonia. Polymyography of cervical muscles, somatosensory evoked potential recordings, and paired transcranial magnetic stimulation were used to assess the dystonic disorder. The results were compared with those obtained in a group of sixteen healthy age matched volunteers. Statistically significant differences between the patient group and the control group were found when the amplitude values of the mean P22/N30 component measured at F [3, 4] and C[3, 4]' electrode positions were compared. The mean amplitude of P22/N30 in both of these electrode positions contralaterally to the direction of head deviation was significantly higher in the patient group (p 60 months) were included. Patients were categorized as asymptomatic if they were free of cerebrovascular incidents for at least 24 months (n = 18). Symptomatic patients had suffered ischemic strokes or transient ischemic attacks within the previous 3 days (n = 35). Platelet function was assessed using the PFA-100 system that allows for quantitative assessment of platelet function, reporting platelet aggregatability as the time required to close a small aperture in a biologically active membrane. RESULTS: Collagen/epinephrine closure times were significantly shorter in symptomatic patients than in asymptomatic patients (p < 0.01). Individual closing times were normal in 12 of 35 symptomatic patients (34 % non-responders) whereas all asymptomatic patients had prolonged closure times. CONCLUSIONS: Aspirin non responder status may contribute to failure of aspirin therapy in the secondary prevention of cerebrovascular incidents in as much as 30-40 % of patients. Quantitative assessment of platelet functions may provide a means to predict aspirin treatment failure in individual patients and to re-direct therapeutic strategies. PMID- 12527995 TI - The normal appearing grey matter in primary progressive multiple sclerosis: a magnetisation transfer imaging study. AB - BACKGROUND: In 10-15 % of patients with multiple sclerosis (MS), the clinical course is characterized by slow progression in disability without relapses (primary progressive (PP) MS). The mechanism of disability in this form of MS is poorly understood. Using magnetization transfer ratio (MTR) imaging, we investigated normal appearing white matter (NAWM) and normal appearing grey matter (NAGM) in PPMS and explored the relationship of MTR measures with disability. METHODS: Thirty patients with PPMS and 30 age matched controls had spin echo based MTR imaging to study lesions and normal appearing tissues. The brain was segmented into NAWM and NAGM using SPM99 with lesions segmented using a semiautomated local thresholding technique. A 75% probability threshold for classification of NAWM and NAGM was used to diminish partial volume effects. From normalized histograms of MTR intensity values, six MTR parameters were measured. Mean lesion MTR and T2 lesion volume were also measured. Disability was assessed using Kurtzke's expanded disability status scale (EDSS). RESULTS: Compared with controls, patients exhibited a significant reduction in mean NAWM (p = 0.001) and NAGM (p = 0.004) MTR. Spearman's rank correlation of EDSS with the six MTR parameters in NAWM and NAGM, mean lesion MTR, and T2 lesion volume, was only significant with mean NAGM MTR (r = -0.41, p = 0.02), the 25th percentile of NAGM MTR intensity (r = -0.37, p = 0.05), and T2 lesion volume (r = 0.39, p = 0.04). Multiple regression analysis of the relationship between EDSS and 4 MR parameters representing each tissue type (mean NAWM MTR, mean NAGM MTR, mean lesion MTR, T2 lesion volume) showed that the association of EDSS with mean NAGM MTR remained significant. CONCLUSIONS: There appear to be significant abnormalities in the NAGM in PP MS. Further investigation of the pathological basis and functional significance of grey matter abnormality in PPMS is warranted. PMID- 12527996 TI - Effect of myasthenic immunoglobulin G on motor end-plate morphology. AB - This study was undertaken to clarify the role of complement in acetylcholine receptor loss and degeneration of the postsynaptic membrane in myasthenia gravis (MG). We examined the end-plate morphology in rats with passively transferred immunoglobulin G (IgG) from myasthenic patients and the effect of complement by treatment of the rats with cobra venom factor. We injected peroxidase-labeled alpha-BuTx (P-BuTx) into the extensor digitorum longus (EDL) muscle to label the motor end-plates. Three hours later, 100 mg of IgG from MG patients or healthy controls was injected into the tail vein. The EDL was removed 48 hours after the injection of IgG. The presence of macrophages and degeneration of the postsynaptic membrane were seen in 4 of 6 IgG samples from MG patients and a decrease in AChRs in the other 2 samples. These changes were reversed completely by treatment with cobra venom factor in all but one case in which the end-plates were severely degenerated. Injection of MG IgG only never induced end-plate morphology changes. The results suggest that complement has a critical role in degeneration of the postsynaptic membrane and AChR loss at the motor end-plates in the passively transferred model and probably in human MG. PMID- 12527997 TI - Periodic leg movements in patients with Parkinson's disease are associated with reduced striatal dopamine transporter binding. AB - We used single photon emission computed tomography (SPECT) to study striatal [(123)I]beta-CIT binding and polysomnography to study periodic leg movements during sleep (PLMS) in eleven patients with idiopathic Parkinson's disease (PD). The reduced striatal [(123)I]beta-CIT binding was significantly correlated with the number of PLMS. We propose that striatal dopaminergic nerve cell loss is involved in the increased number of PLMS in PD patients. PMID- 12527998 TI - Leber's hereditary optic neuropathy mutations in ethambutol-induced optic neuropathy. AB - Primary mitochondrial DNA (mtDNA) mutation at the nt 11778 site in Leber's hereditary optic neuropathy (LHON) has been reported to be present in patients with ethambutol-induced optic neuropathy. To study further this association between LHON and ethambutol-induced optic neuropathy, we tested ethambutol induced optic neuropathy patients for the presence of the mtDNA mutations at nucleotides (nt)-11778, nt-14484, nt-3460, nt-15257, nt-9438, nt-9804, nt-13730, and nt-14459 in 24, 15, 8, 6, 5, 5, 5, and 5 patients respectively. However, none of the ethambutol-induced optic neuropathy patients was found to exhibit any pathogenic LHON mtDNA mutation. In conclusion, we found no evidence of any association between ethambutol-induced optic neuropathy and the LHON mutations. PMID- 12527999 TI - Ropinirole as an adjunct to levodopa in the treatment of Parkinson's disease: a 16-week bromocriptine controlled study. AB - BACKGROUND: and objectives Ropinirole is a non-ergoline, selective dopamine D(2) agonist. The aim of this study was to evaluate the efficacy and safety of ropinirole as an adjunct to levodopa in the treatment of Parkinson's disease (PD) complicated by motor fluctuations. METHODS: A total of 76 patients with PD (Hoehn and Yahr stage II to IV) were included in this trial. Each patient was randomly allocated to receive either ropinirole (n = 37) or bromocriptine (n = 39) as an adjunct to levodopa over a 16-week period. Ropinirole and bromocriptine were titrated for optimal efficacy and tolerability. This optimal dose was then maintained for the rest of the study. Response rate was defined as the percentage of patients who achieved at least a 20 % reduction in levodopa dose. Clinical status was also assessed using the Unified Parkinson's Disease Rating Scale (UPDRS), Clinical Global Impression (CGI), and reduction in time spent 'off'. RESULTS: Ropinirole produced a significantly greater response rate than bromocriptine (odds ratio 2.995, 95 % C. I. (1.157, 7.751) p < 0.05). There was also a statistically significant difference between the groups in the proportion of patients who were 'improved' on the CGI improvement scale (91.9 % for ropinirole, 74.3 % for bromocriptine, p = 0.046). Other measures, including at least a 20 % improvement in the UPDRS motor score (70 % for ropinirole and 63.3 % for bromocriptine), and a 20 % reduction in 'off' duration (81 % for ropinirole and 52.4 % for bromocriptine) showed a trend in favour of ropinirole. There was no significant difference between the two groups in the overall incidence of adverse effects (ropinirole, 59.5 %; bromocriptine, 59 %). In each group, the most common side-effects were dizziness, dyskinesia and nausea/vomiting. No patients were withdrawn from the study because of side-effects. CONCLUSION: Ropinirole was found to be safe and well-tolerated. Ropinirole as an adjunct to levodopa in the treatment of PD with motor fluctuation was associated with more significant reduction of levodopa dose and, on one form of analysis, with significantly greater improvement in CGI ratings than bromocriptine. On the other efficacy measures the two drugs were comparable. PMID- 12528000 TI - Ropinirole as an adjunct to levodopa in the treatment of Parkinson's disease A short commentary. PMID- 12528001 TI - Focal hypertrophic cranial pachymeningitis associated with temporal arteritis: a new case report. PMID- 12528002 TI - Loss of color vision during long-term treatment with pramipexole. PMID- 12528003 TI - Catatonia and Parkinsonism due to extrapontine myelinolysis following rapid correction of hyponatremia: a case report. PMID- 12528004 TI - Subacute syphilitic meningomyelitis with characteristic spinal MRI findings. PMID- 12528005 TI - HIV-1 related ischaemic trochlear nerve palsy. PMID- 12528006 TI - Cataplexy in association with Moebius syndrome. PMID- 12528007 TI - Hodgkins disease presenting with granulomatous angiitis of the central nervous system. PMID- 12528008 TI - Hypokalemic myopathy due to ingestion of earth. PMID- 12528009 TI - Reversible angiopathy and encephalopathy after blood transfusion. PMID- 12528010 TI - Woldag H, Hummelsheim H: evidence-based physiotherapeutic concepts for improving arm and hand function in stroke patients: a review J Neurol (2002) 249:518-528. PMID- 12528011 TI - Paul Nayrac (1899-1973). PMID- 12528012 TI - Effects of cannabinoids on lithium-induced conditioned rejection reactions in a rat model of nausea. AB - RATIONALE: Marijuana has been reported to suppress nausea produced by chemotherapy treatment in human cancer patients. Although there is abundant evidence that cannabinoid agonists attenuate vomiting in emetic species, there has been little experimental evidence of their anti-nausea potential. Considerable evidence suggests that conditioned rejection reactions in rats reflect nausea. The present experiments evaluated the potential of low doses of the cannabinoid agonists, delta-9-tetrahydrocannabinol (THC; 0.5 mg/kg, i.p.), and HU-210 (0.001 mg/kg and 0.01 mg/kg, i.p.), and the CB(1) antagonist SR 141716A in modulating the establishment and the expression of lithium-induced conditioned rejection reactions in rats. OBJECTIVES: To evaluate the effect of cannabinoids on conditioned rejection reactions, a rat model of nausea. METHODS: In experiments 1 and 2, respectively, rats were injected with cannabinoid agonists, THC (0.5 mg/kg, i.p.) and HU-210 (0.001, 0.005 or 0.01 mg/kg), 30 min prior to exposure to 0.1% saccharin solution by intraoral infusion. Immediately following saccharin exposure, they were injected with 20 ml/kg 0.15 M lithium chloride or saline. On each of two test trials, the rats were injected with the cannabinoid or vehicle 30 min prior to exposure to saccharin. In experiment 3, rats were injected with the CB(1) antagonist, SR-141716A (2.5 mg/kg) or a combination of SR-141716A and HU-210 (0.01 mg/kg) 30 min prior to an infusion of saccharin followed by injection of lithium or saline. They were given a single drug-free test trial. Experiment 4 replicated and extended the findings of experiment 3. RESULTS: delta-9-THC and HU-210 interfered with the establishment and the expression of lithium-induced conditioned rejection reactions. The suppressive effect of HU-210 on rejection reactions was reversed by pretreatment with SR-141716A. Administration of SR-141716A prior to conditioning potentiated lithium-induced conditioned rejection reactions. CONCLUSIONS: These results indicate that the establishment and the expression of lithium-induced conditioned rejection reactions are suppressed by pretreatment with cannabinoid agents. These effects appear to be mediated by their action on the CB(1) receptor, because they are reversed by pretreatment with SR-141716A. Finally, our results suggest that endogenous cannabinoids play a role in modulation of nausea, because the antagonist potentiated lithium-induced nausea. PMID- 12528014 TI - Legal implications of clinical practice guidelines. PMID- 12528013 TI - The hidden faces of the epidemiology of antibiotic resistance. PMID- 12528016 TI - Adenosine triphosphate-magnesium chloride: relevance for intensive care. AB - BACKGROUND: Despite aggressive resuscitation shock often results in multiple organ failure characterized by increased energy demands of organs and decreased ability of effective energy production. The administration of ATP-MgCl(2) as a supportive measure has been investigated in various animal models of ischemia/reperfusion injury and hemorrhagic, endotoxic, and septic shock. INVESTIGATIONS: These studies showed improvement in organ blood flow, microcirculation, energy balance, cellular and mitochondrial, functions and restoration of immune competence, ultimately leading to increased survival. Originally these effects were attributed to direct energy provision by the ATP-Mg complex, but the minute amount of ATP infused compared to the body's ATP formation rate suggests that other mechanisms must be responsible for its beneficial properties such as stabilization of the cell membrane, phosphorylation of membrane proteins, decreased cell swelling, and improved microcirculatory perfusion. CONCLUSIONS: The experimental evidence currently available suggests the use of ATP-MgCl(2) as a therapeutic adjunct in patients with multiple-organ dysfunction. In addition, given the extremely short half-life which allows both rapid titration and control of the systemic hemodynamic response, for example, reduction in mean arterial pressure, ATP-MgCl(2) may be suitable as an alternative to other fast-acting vasodilators used for the management of acute pulmonary hypertensive crises and/or for the maintenance blood pressure during aortic cross-clamping. PMID- 12528017 TI - Pulmonary artery occlusion pressure. PMID- 12528018 TI - Nosocomial pneumonia. AB - Nosocomial pneumonia, or terminal pneumonia as it was formerly called, results from the repetitive microaspiration of contaminated oropharyngeal secretions into the lungs in the presence of impaired host defenses. This pathophysiologic sequence was suggested by the observations of Osler but clarified by the seminal work of Rouby and colleagues. The enormous impact of antimicrobial agents on the organisms responsible for nosocomial pneumonias was first identified by Kneeland and Price who found that organisms of the normal pharyngeal flora virtually disappeared in terminal pneumonias following administration of these drugs, being replaced by gram-negative bacilli. The remarkable susceptibility of seriously ill patients to becoming colonized by exogenous organisms, even in the absence of antimicrobial therapy, was shown by Johanson et al. These factors, antibiotics and the change in bacterial binding receptors in the airways associated with illness, lead to infections caused by exogenous organisms that are frequently resistant to antimicrobial agents. Clinical findings that usually identify patients with respiratory infections are unreliable for the diagnosis of nosocomial pneumonias as shown by Andrews et al. Invasive techniques, especially the protected specimen brush (PSB) technique, avoid contamination of the specimen by proximal secretions and accurately reflect the bacterial burden of the lung, as first shown by Chastre et al. Quantitation of such specimens serves as an excellent proxy for direct cultures of the lung and are the current gold standard for diagnosis. PMID- 12528019 TI - Intra-abdominal pressure monitoring in liver transplant recipients: a prospective study. AB - OBJECTIVE: No data are available on the incidence and clinical relevance of increased intra-abdominal pressure after orthotopic liver transplantation. This study assessed abdominal hypertension in a population of transplanted patients as this may be an important cofactor in early postoperative complications. DESIGN AND SETTING: Prospective clinical study in an intensive care unit of a national health system teaching hospital PATIENTS AND MEASUREMENTS: Abdominal pressure was measured every 6 h using the urinary bladder method and was considered elevated when it was 25 mmHg or higher. Hemodynamic status was evaluated at the same times. Renal function was assessed on the basis of hourly urinary output by calculating serum creatinine on postoperative days 2 and 4 and the filtration gradient; patient outcomes were also considered. RESULTS: Intra-abdominal hypertension was observed in 32% of cases; the subjects with high abdominal pressure had significantly lower mean artery pressure values but did not differ in terms of central venous pressure or cardiac output. High intra-abdominal pressure was more frequently associated with renal failure, a lower filtration gradient, delayed postsurgical weaning from ventilation, and a worse outcome. CONCLUSIONS: Abdominal hypertension is frequent after liver transplantation and may be associated with a complicated post-operative course. PMID- 12528020 TI - End-expiratory lung impedance change enables bedside monitoring of end-expiratory lung volume change. AB - OBJECTIVE: The aim of the study was to investigate the effect of lung volume changes on end-expiratory lung impedance change (ELIC) in mechanically ventilated patients, since we hypothesized that ELIC may be a suitable parameter to monitor lung volume change at the bedside. DESIGN: Clinical trial on patients requiring mechanical ventilation. SETTINGS: Intensive care units of a university hospital. PATIENTS: Ten mechanically ventilated patients were included in the study. INTERVENTION: Patients were ventilated in volume-controlled mode with constant flow and respiratory rate. In order to induce changes in the end-expiratory lung volume (EELV), PEEP levels were increased from 0 mbar to 5 mbar, 10 mbar, and 15 mbar. At each PEEP level EELV was measured by an open-circuit nitrogen washout manoeuvre and ELIC was measured simultaneously using Electrical Impedance Tomography (EIT) with sixteen electrodes placed on the circumference of the thorax and connected with an EIT device. Cross-sectional electro-tomographic measurements of the thorax were performed at each PEEP level, and a modified Sheffield back-projection was used to reconstruct images of the lung impedance. ELIC was calculated as the average of the end-expiratory lung impedance change. RESULTS. Increasing PEEP stepwise from 0 mbar to 15 mbar resulted in an linear increase of EELV and ELIC according to the equation: y =0.98 x -0.68, r(2)=0.95. CONCLUSION: EIT is a simple bedside technique which enables monitor lung volume changes during ventilatory manoeuvres such as PEEP changes. PMID- 12528021 TI - Thyroid hormone response to moderate hypothermia in severe brain injury. AB - OBJECTIVE: To examine the effect of controlled moderate hypothermia on thyroid response in head-injured patients. DESIGN: Prospective, controlled, randomized study. SETTING: University hospital intensive care unit (ICU). PATIENTS: Twenty eight patients with severe blunt head injury (Glasgow Coma Scale < or =9). INTERVENTION: Patients were randomly assigned to a hypothermia or a normothermia group. Hypothermia (32-33 degrees C) was induced within 8 h after trauma and maintained for a mean of 36 h. All patients were sedated and mechanically ventilated. MEASUREMENTS AND RESULTS: Thyroid-stimulating hormone( TSH), free and total triiodothyronine (FT3/TT3), reverse triiodothyronine (RT3) and thyroxine (FT4/TT4) were measured during the hypothermia or corresponding normothermia period, after regaining normothermia and 4-6 days later. Of 28 patients included in the study, 11 subjects were treated with hypothermia and 13 patients with normothermia. Four patients had to be excluded. In both groups, serum concentrations of TT3 and FT3 were just below the lower normal range whereas RT3 serum concentrations were near the upper limit of the normal range. TSH serum concentrations were not increased. No statistically significant intra- or inter group differences were observed. CONCLUSIONS: Thyroid hormone patterns during moderate hypothermia in head-injured patients did not differ from the well known "low T3 state" which is observed in other forms of severe illness. PMID- 12528022 TI - Rational use of antibiotics in the intensive care unit: impact on microbial resistance and costs. AB - OBJECTIVE: To evaluate the impact of an intensive care unit (ICU) antibiotic-use policy on the microbial resistance in nosocomial infections and costs. DESIGN: Comparative study before and after policy implementation. SETTINGS: An eleven-bed ICU in a general hospital. PATIENTS: All patients admitted for at least 48 h during a 5year period (1994-1998). INTERVENTIONS: In 1995, implementation of an antibiotic-use policy. MEASUREMENTS AND MAIN RESULTS: Patients' general characteristics, incidence of nosocomial infections, antibiotic-selective pressure (the number of days of antibiotic treatment for 1,000 days of presence in the ICU), presence and types of multi-resistant micro-organisms and costs linked to antibiotic use were recorded before (1994) and after implementation of the policy (1995-1998). For each year, patients' general characteristics and the incidence of nosocomial infections were the same. Costs linked to antibiotics use showed a progressive reduction (100% for 1994, 81% for 1995, 65% for 1998). Antibiotic-selective pressure diminished (from 940 days of antibiotic use per 1,000 days (1994) to 610 (1998), p<10(-5)). A statistically significant reduction in nosocomial infections due to antimicrobial resistant micro-organisms was observed (from 37% (1994) to 15% (1998) of nosocomial infections, p<10(-5)) after 3 years of implementation of the policy, essentially due to a reduction in methicillin-resistant Staphylococcus aureus and ceftriaxone-resistant Enterobacteriaceae. Nosocomial infections due to ceftazidime-resistant Pseudomonas species or extended-spectrum ss-lactamase Enterobacteriaceae showed no reduction. CONCLUSIONS: Antibiotic-use policy allowed a reduction in antibiotic-selective pressure, costs linked to antibiotics and selective reduction of nosocomial infections due to antimicrobial resistant micro organisms. PMID- 12528023 TI - Early identification of sepsis and mortality risks through simple, rapid clot waveform analysis. Implications of lipoprotein-complexed C reactive protein formation. AB - OBJECTIVE: To determine if the rapid waveform profile of the activated partial thromboplastin time (aPTT) assay, which detects lipoprotein-complexed C reactive protein (LCCRP) formation, predicts sepsis and mortality in critically ill patients. DESIGN: Observational, cohort study. SETTING: General intensive therapy unit (ITU) of a tertiary care hospital. PATIENTS AND PARTICIPANTS: A total of 1187 consecutive patients admitted to the ITU. INTERVENTION: Activated partial thromboplastin time transmittance waveform analysis was performed within the first hour of admission to the ITU. The degree of change causing a biphasic waveform was quantified through the drop in light transmittance level. MEASUREMENTS AND RESULTS: Three hundred forty-six patients had a biphasic waveform on admission to the ITU with a mortality rate of 44% compared with 26% for those with normal waveforms. Logistic regression models showed direct correlation between the likelihood for sepsis and in-patient mortality with increasing waveform abnormalities. The mortality fraction was 0.3 with normal waveforms versus 0.6 when the light transmittance decreased by 30%. The odds ratio (OR) for mortality and sepsis were 4.5 and 11, respectively, from the most abnormal to normal aPTT waveforms. These were comparable with APACHE II scores and superior to those estimated by CRP for mortality (OR 2.3) / sepsis (OR 6.4) prediction. CONCLUSION: Waveform analysis within the first hour of ITU admission is a single, simple and rapid method of identifying the risks of mortality and sepsis. Its measure of LCCRP formation shows superior prediction over CRP alone and it warrants further assessment as a tool to triage and target prompt, appropriate treatment in the ITU. PMID- 12528024 TI - Hemoglobin-based oxygen carriers do not alter platelet functions: study of three chemically modified hemoglobin solutions. AB - OBJECTIVE: Chemically modified hemoglobins are being developed as potential oxygen-carrying blood substitutes (HBOCs). Clinical and preclinical data demonstrate the vasoactive properties of HBOCs by trapping of nitric oxide, which is also known to have platelet inhibitory activities properties. This study evaluated the effects of three structurally different HBOCs (Hb-Dex-BTC, alphaalpha-Hb, and o-raffinose-poly-Hb) on platelet functions in vitro to compare to those elicited by plasma substitutes, such as hydroxyethylstarch. DESIGN: Platelet activation state was assessed using platelet-rich plasma diluted to 20% (v/v) with the different solutions, by main measuring glycoproteins (GPIb, GPIIb/IIIa, and P-selectin) using flow cytometry. Aggregation was assessed by impedance aggregometry on whole blood hemodiluted to 20% (v/v) with the solutions. SETTING: Biological hematology department of the university hospital of Nancy-Brabois. PATIENTS AND PARTICIPANTS: Ten healthy volunteers consent and informed of the study who denied taking any drugs at the time of the experiment. MEASUREMENTS AND RESULTS: None of these solutions induced activation nor modified reactivity of platelets as measured by the surface expression of glycoproteins GPIb, GPIIb/IIIa, and P-selectin. Moreover, none of these solutions induced platelet aggregation when added alone, nor modified the aggregation patterns of platelets induced by collagen (0.5 microg/ml) and thrombin receptor agonist peptide (12.5 microM). CONCLUSIONS: The three tested structurally different HBOCs, as with hydroxyethylstarch, did not alter platelet functions in vitro. PMID- 12528025 TI - Post-extubation stridor in intensive care unit patients. Risk factors evaluation and importance of the cuff-leak test. AB - OBJECTIVE: To evaluate the incidence and identify factors associated with the occurrence of post-extubation stridor and to evaluate the performance of the cuff leak test in detecting this complication. DESIGN: Prospective, clinical investigation. SETTING: Intensive care unit of a university hospital. PATIENTS: Hundred twelve extubations were analyzed in 112 patients during a 14-month period. INTERVENTION: A cuff-leak test before each extubation. MEASUREMENTS AND RESULTS: The incidence of stridor was 12%. When we chose the thresholds of 130 ml and 12% to quantify the cuff-leak volume, the sensitivity and the specificity of the test were, respectively, 85% and 95%. The patients who developed stridor had a cuff leak significantly lower than the others, expressed in absolute values (372+/-170 vs 59+/-92 ml, p<0.001) or in relative values (56+/-20 vs 9+/-13%, p<0.001). Stridor was associated with an elevated Simplified Acute Physiology Score (SAPS II), a medical reason for admission, a traumatic or difficult intubation, a history of self-extubation, an over-inflated balloon cuff at admission to ICU and a prolonged period of intubation. These results provide a framework with which to identify patients at risk of developing a stridor after extubation. CONCLUSION: A low cuff-leak volume (<130 ml or 12%) around the endotracheal tube prior to extubation is useful in identifying patients at risk for post-extubation stridor. PMID- 12528026 TI - Decision-making in the ICU: perspectives of the substitute decision-maker. AB - OBJECTIVE: To describe the substitute decision-makers' perspectives related to decision-making in the intensive care unit (ICU) and to determine those variables associated with their overall satisfaction with decision-making. DESIGN: Prospective, multicenter, cohort study. SETTING: Six Canadian university affiliated ICUs. PATIENTS AND PARTICIPANTS: We distributed a validated, self administered questionnaire assessing 21 key aspects of communication and decision making to substitute decision-makers of ICU patients who were mechanically ventilated for more than 48 h. INTERVENTION: None. MEASUREMENTS AND RESULTS: A group consisting of 1,123 substitute decision-makers received questionnaires; 789 were returned (70.3% response rate). Respondents were most satisfied with the frequency of communication with nurses and least satisfied with the frequency of communication with physicians. In terms of overall satisfaction with decision making, 560 (70.9%) of the respondents were either completely or very satisfied. The majority (81.2%) of respondents preferred some form of shared decision-making process. Factors contributing the most to satisfaction with decision-making included: complete satisfaction with level of health care the patient received, completeness of information received, and feeling supported through the decision making process. Satisfaction with decision-making varied significantly across sites. CONCLUSIONS: In this multicenter observational study, we found that most substitute decision-makers for ICU patients wanted to share decision-making responsibility with physicians and that, overall, they were satisfied with their decision-making experience. Adequate communication, feeling supported, and achieving the appropriate level of care for their family member were key determinants of satisfaction with decision-making in the ICU. PMID- 12528027 TI - Intensive care information system reduces documentation time of the nurses after cardiothoracic surgery. AB - OBJECTIVE: Nowadays, registration of patient data on paper is gradually being replaced by registration using an intensive care information system (ICIS). The aim of this study was to evaluate the effect of the use of an ICIS on nursing activity. DESIGN: Randomized controlled trial with a crossover design. SETTING: An 18-bed medical-surgical ICU in a teaching hospital. PATIENTS, NURSES AND INTERVENTIONS: During a 6week period 145 consecutive adult patients admitted to the ICU after uncomplicated cardiothoracic surgery were randomized into two groups: for one group the documentation was carried out using a paper-based registration (Paper), in the second group an ICIS was used for documentation. MEASUREMENTS AND RESULTS: The nursing activities for these patients were studied during two separate periods: the admission period and the registration phase (the period directly following the admission procedure). The duration of the admission procedure was measured by time-motion analysis and the nursing activities in the registration phase were studied by work sampling methodology. All nursing activities during the registration phase were grouped in four main categories: patient care, documentation, unit-related and personal time. The duration of the admission procedure was longer in the ICIS group (18.1+/-4.1 versus 16.8+/-3.1 min, p<0.05). In the registration phase, a 30% reduction in documentation time (Paper 20.5% of total nursing time versus ICIS 14.4%, p<0.001), corresponding to 29 min (per 8h nursing shift) was achieved. This time was completely re-allocated to patient care. CONCLUSIONS: The use of the present ICIS in patients after cardiothoracic surgery alters nursing activity; it reduces the time for documentation and increases the time devoted to patient care. ELECTRONIC SUPPLEMENTARY MATERIAL: is available if you access this article at http://dx.org/10.1007/s00134-002-1542-9. On that page (frame on the left side), a link takes you directly to the supplementary material. PMID- 12528028 TI - Prognosis of child recipients of hematopoietic stem cell transplantation requiring intensive care. AB - OBJECTIVE: To determine prognostic factors in child recipients of hematopoietic stem cell transplantation from blood or bone marrow (BMT) requiring critical care. DESIGN: Retrospective study of a cohort of patients. SETTING: Pediatric Intensive Care Unit (PICU) in a university tertiary care center. PATIENTS AND PARTICIPANTS: Child recipients of BMT requiring PICU admission. MEASUREMENTS AND RESULTS: Of the 151 children receiving transplants in our institution, 44 (29.1%) had 49 admissions to the PICU. Mechanical ventilation (MV) was required in 34 patients (69.4% of all admissions). Overall mortality was 31/44 (70.4%). Mortality in patients requiring MV and not requiring MV was 26/34 (76.5%) and 5/10 (50%), respectively. The following variables were significantly associated with mortality in the univariate analysis: male gender (P=0.02), older age (P=0.03), acute graft versus host disease (aGVHD) grades III or IV (P=0.01), severe hemorrhagic cystitis (P=0.01), the diagnosis of lung injury (P=0.04), the need for MV (P=0.03) or for renal replacement therapy (P=0.02), the presence of respiratory (P=0.003), cardiovascular (P=0.009) or gastrointestinal (P=0.01) failures, and the failure of > or =3 organs (P=0.01). In the multivariate analysis, the presence of aGVHD grades III or IV, male gender, severe hemorrhagic cystitis, and the failure of > or =3 organs were found to be independent predictors of mortality. CONCLUSIONS: The need for intensive care is common among child recipients of a BMT. These patients have a high mortality rate but some complications are reversible with critical care support. Certain clinical parameters are useful to establish a realistic prognosis and to optimize the use of the available resources. PMID- 12528029 TI - Outcome of children with different accessibility to tertiary pediatric intensive care in a developing country--a prospective cohort study. AB - OBJECTIVE: Lack of direct access to tertiary pediatric intensive care services in rural hospitals may be associated with poorer outcome among critically ill children. Inter-hospital transport by non-specialized teams may also lead to increased morbidity and even mortality. We therefore studied the outcome of children with different accessibility to tertiary pediatric care in Malaysia. METHODS: We prospectively compared the Pediatric Risk of Mortality (PRISM II) adjusted standardized mortality ratio (SMR), unanticipated deaths and length of stay of 131 patients transported from rural hospitals (limited access) with 215 transferred from the casualty wards or other in-hospital wards (direct access) to a tertiary pediatric ICU. RESULTS: The transported patients were younger than the in-hospital patients (median age 1.0 versus 6.0 months, p=0.000) and were more likely to have respiratory diseases. Other baseline characteristics did not differ significantly. Differences in access to tertiary intensive care from community hospitals was associated with an extended median length of stay (4.0 versus 2.0 days, p=0.000) but did not affect SMR (0.92 versus 0.84, rate ratio 1.09, 95% CI 0.57-2.01; p=0.348) or percentage of unexpected deaths (4.8% versus 2.8%, p=0.485). The adjusted odds ratio for mortality (1.7, 95% CI 0.7-4.3) associated with transfer was not statistically significant (p=0.248). CONCLUSIONS: The outcome of critically ill children transferred from community hospitals did not differ from that of those who develop ICU needs in the wards of a tertiary center, despite being transported by non-specialized teams. Outcome was not affected by initial inaccessibility to intensive care if the children finally received care in a tertiary center. PMID- 12528030 TI - Effects of controlled mechanical ventilation on respiratory muscle contractile properties in rabbits. AB - OBJECTIVE: We examined in rabbits the effects of more than 48 h of mechanical ventilation on the contractile properties and fiber type adaptations of the respiratory muscles. DESIGN AND SETTING: Experimental prospective study in a university laboratory. ANIMALS AND INTERVENTIONS: Nineteen rabbits were randomly allocated to two groups: control (n=10) or mechanically ventilated (MV; n=9) for 51+/-3 h. MEASUREMENTS AND RESULTS: Respiratory muscles contractile properties were analyzed before and after a fatigue protocol using in vivo isometric 1-s tetanic contraction characteristics in both muscles: peak tetanic force, contraction time, relaxation time, and total contraction time. Both muscle fiber type proportions, diameter, and cross-sectional areas were measured using ATPase staining. The MV rabbits showed significant weight loss in both muscles, accompanied by a reduced peak tetanic force (9.96+/-3.2 vs. 7.44+/-2.2 N for diaphragm of control and MV animals respectively), fatigue resistance index, and increased relaxation time (57.5+/-8.7 vs. 85.8+/-9.4 ms for diaphragm of control and MV animals) and contraction time. These impairments in the MV group worsened after the fatigue runs. Both muscle showed a significant atrophy of type IIa and IIb fibers but a stability in type I fibers cross-sectional area. CONCLUSIONS: Mechanical ventilation in rabbits produces alterations in contractile properties of the diaphragm and 5th external intercostal muscle, increases both muscles fatigue, and promotes atrophy of type II fibers. PMID- 12528031 TI - Effects of prolonged mechanical ventilation on respiratory muscle ultrastructure and mitochondrial respiration in rabbits. AB - OBJECTIVE: To investigate in rabbits whether prolonged mechanical ventilation (PMV) leads to ultrastructural changes in respiratory muscles and alters diaphragm mitochondrial respiration. DESIGN AND SETTING: Experimental prospective study in a university laboratory. ANIMALS AND INTERVENTIONS: We studied respiratory muscles of seven rabbits after 49+/-1 h of controlled mechanical ventilation. Ten nonventilated rabbits were used as a control group. MEASUREMENTS AND RESULTS: After mechanical ventilation electron-microscopic observations of the diaphragm and the external intercostal muscles revealed disrupted myofibrils, increased number of lipid vacuoles in the sarcoplasm, and smaller mitochondria with focal membrane disruptions. Volumetric and numerical densities of the mitochondria were significantly lower in the PMV group than the control group. Mitochondrial respiration was quantified in isolated diaphragm muscle-cell mitochondria using two respiratory substrates. There was no difference in oxygen consumption values in the three states of mitochondrial respiration between the two groups except for state 2 (basal state) with pyruvate/malate parameter (53.5+/-20 for the ventilated group vs. 33.8+/-10.2 nmol atom O/mg per minute for the control group). There was no significant difference between groups in ADP/O ratio or respiratory control ratio. CONCLUSIONS: PMV leads to respiratory muscle cell degeneration and minor changes in oxidative phosphorylation coupling in diaphragmatic mitochondria. These phenomena may mediate part of damage of respiratory muscles after inactivity related to PMV. PMID- 12528032 TI - An improved porcine model of stable methacholine-induced bronchospasm. AB - OBJECTIVE: To validate an animal model replicating the pathophysiological characteristics of severe induced bronchospasm observed in humans, with a high level of stability permitting measurements such as the assessment of ventilation perfusion relationships with the multiple inert gas elimination technique. DESIGN AND SETTING: Experimental study in an animal research laboratory. SUBJECTS: 13 piglets (age 3-4 months) were studied and 7 underwent the complete protocol INTERVENTIONS: The animals were anesthetized and paralyzed. Mechanical ventilation was initiated in a volume-controlled mode. Ventilatory parameters were adjusted to obtain normocapnia and were maintained constant during the bronchospasm. Methacholine was administered via a synchronized nebulizer and progressively adjusted to obtain a stable twofold increase in peak inspiratory pressure. MEASUREMENTS AND RESULTS: Cardiopulmonary physiological data including assessment of lung mechanics and measurement of ventilation-perfusion relationships were obtained before and during the bronchospasm. Peak inspiratory pressure increased from 19.7+/-2.9 to 44.4+/-7.1 cmH(2)O during the bronchospasm. The latter remained stable over 2 h. Respiratory mechanics, gas exchange, and ventilation-perfusion distribution changes typical of those observed in severe bronchospasm in humans were observed in all animals. CONCLUSIONS: The present experimental model replicates some of the physiopathological characteristics of severe human bronchospasm, and its stability should facilitate studies of the effects of different ventilatory modes in the setting of acute severe asthma. PMID- 12528033 TI - Fiberoptic bronchoscopy during noninvasive positive pressure ventilation delivered by helmet. AB - OBJECTIVE: To evaluate the feasibility and safety of fiberoptic bronchoscopy (FOB) with bronchoalveolar lavage (BAL) during noninvasive positive pressure ventilation (NPPV) delivered by helmet in patients with acute respiratory failure (ARF) and suspected pneumonia. DESIGN AND SETTING: Prospective, clinical investigation in a general intensive care unit (ICU) of a university hospital. PATIENTS AND PARTICIPANTS: Four adult patients with ARF who underwent NPPV via the helmet and required fiberoptic BAL for suspected pneumonia. INTERVENTIONS: NPPV was delivered through the helmet in the pressure support ventilation mode. The specific seal connector placed in the plastic ring of the helmet allowed the passage of the bronchoscope, maintaining assisted ventilation. Arterial blood gas levels, pH, oxygen saturation, respiratory rate, heart rate, and mean arterial blood pressure were monitored during the study. RESULTS: Helmet NPPV avoided gas exchanges deterioration during FOB and BAL, with good tolerance. During the procedure heart rate increased by 5% and mean arterial blood pressure by 7% over baseline; these levels returned to prebronchoscopic values immediately after the withdrawal of the bronchoscope. Endotracheal intubation was never required during the 24 h after the procedure. BAL yielded diagnostic information in three of four patients. CONCLUSIONS: NPPV through the helmet allows a safe diagnostic FOB with BAL in patients with hypoxemic ARF, avoiding gas exchange deterioration, and endotracheal intubation. ELECTRONIC SUPPLEMENTARY MATERIAL: is available if you access this article at http://dx.doi.org/10.1007/s00134-002-1554-5. On that page (frame on the left side), a link takes you directly to the supplementary materials. PMID- 12528034 TI - Effect of end-inspiratory pause duration on plateau pressure in mechanically ventilated patients. AB - OBJECTIVE: To compare the values of plateau pressure (Pplat) recorded at different times after end-inspiratory occlusion and those of static elastance (Est,rs) and total resistance (Rrs) of the respiratory system. DESIGN: Physiological study. SETTING: Medical intensive care unit of a university hospital. PATIENTS: Eleven patients with ARDS and ten patients with COPD requiring tracheal intubation and mechanical ventilation were investigated. COPD patients were investigated on zero end-expiratory pressure (ZEEP) and ARDS patients on both ZEEP and positive end-expiratory pressure (PEEP). MEASUREMENTS AND RESULTS: Respiratory mechanics were assessed using the rapid airway occlusion technique. Tracheal pressure (Ptr) was measured downstream the endotracheal tube. Ptr was recorded 0.5 s, 1 s, 2 s, 3 s, and 5 s after a 5-s end-inspiratory occlusion. Est,rs and Rrs were computed at the same times using standard formula. In ARDS patients on ZEEP, Pplat amounted to 20+/-5, 20+/-5, 19+/-5, 19+/-5, and 18+/-5 cmH(2)O at 0.5, 1, 2, 3 and 5 s, respectively (P <0.001). In COPD patients, these values were 18+/-4 cmH(2)O, 17+/-4 cmH(2)O, 17+/-4 cmH(2)O, 16+/ 4 cmH(2)O, and 16+/-4 cmH(2)O (P <0.001). Except for one ARDS patient on PEEP, Pplat was always less than 35 cmH(2)O, regardless of the time of measurement. As compared to 5 s, measurements at 0.5 s resulted in overestimation of Est,rs by 14% and 29% and in underestimation of Rrs by 34% and 24%, in ARDS and COPD patients, respectively. CONCLUSIONS: Very early post-occlusion values of Pplat were statistically greater than at 3 s or 5 s. This probably has no major impact on the occurrence of volutrauma. Clinicians must be aware, however, that Est,rs and Rrs are greatly modified by the time of recording of Pplat. PMID- 12528035 TI - Neutropenia and the development of the systemic inflammatory response syndrome. AB - OBJECTIVE: To determine outcome and natural course of systemic inflammatory response syndrome (SIRS) stages in adult febrile neutropenic patients. DESIGN AND SETTING: Retrospective cohort study in the medicine department and intensive care unit of a university hospital. PATIENT: Adults with cancer-related neutropenia and community-acquired fever. MEASUREMENTS AND RESULTS: Patients were classified on admission according to SIRS parameters, tumor type, and degree of neutropenia. Records of clinical and laboratory data during hospitalization were reviewed. Univariate and logistic regression analyses were performed. Seventy-nine events in 62 patients were analyzed. Overall mortality rate was 20.2% (16/79). Mortality increased as SIRS stage worsened on admission. No patients with stage 2 SIRS died (neutropenia and fever alone) but 11.1% of patients with SIRS 3, 43.4% with SIRS 4, 66.6% with sepsis induced hypotension, and 90% with septic shock. SIRS stage on admission was an independent predictor of death and was related directly to rate of progression to shock, i.e., none of the patients with SIRS 2, 2.7%(1/36) of those with SIRS 3, and 30.4% (7/23) of those admitted with SIRS 4. CONCLUSIONS: Mortality and progression to septic shock increased as more SIRS criteria were met on admission. SIRS stages could serve as a risk-assessing model in febrile neutropenic patients. PMID- 12528036 TI - Cost-effectiveness of an antiseptic-impregnated central venous catheter in the ICU. PMID- 12528037 TI - Respiratory failure due to pharyngeal dysfunction: a singular presentation of brainstem listeriosis. PMID- 12528038 TI - Tube thoracostomy--abandon the trocar. PMID- 12528040 TI - First manifestation of a hereditary homozygous factor VII deficiency in a 3-year old girl suspected as purpura fulminans. PMID- 12528041 TI - Ascorbic acid is decreased in induced sputum of mild asthmatics. AB - Asthma is primarily an airways inflammatory disease, and the bronchial airways have been shown to be particularly susceptible to oxidant-induced tissue damage. The antioxidant ascorbic acid (AA) plays an essential role in defending against oxidant attack in the airways. Decreased levels of AA have been reported in the plasma and BAL fluid of asthmatics, but not at the site directly proximal to asthma pathology, the bronchial airways. We investigated whether asthmatics have deficient levels of AA in the airways compared to healthy subjects. We performed induced sputum (IS) in a group of mild asthmatics (n = 16) and healthy controls (n = 18) in order to compare constitutive levels of antioxidants in the airways of these two groups. We report that asthmatics had significantly decreased AA in both the cellular (17 +/- 3 ng/10(6) cells vs. 40 +/- 4 ng/10(6) cells) and fluid phase fraction (616 +/- 152 ng/ml vs. 937 +/- 161 ng/ml) of the IS sample compared to normals. No differences were found with glutathione (GSH) and alpha tocopherol. These results suggest that AA deficiency may be either an underlying factor in the pathophysiology of asthma or a response to asthmatic airways inflammation. PMID- 12528042 TI - Development of a high-volume concentrated ambient particles system (CAPS) for human and animal inhalation toxicological studies. AB - A two-stage, high-volume, ambient particle concentrator was developed and characterized. This versatile system, depending on its operational parameters, can be used to fractionate and concentrate particles in three size ranges (PM(10 2.5), PM(10-1), PM(2.5-1)). The performance of this concentrated ambient particle system (CAPS), as well as its individual virtual impaction stages, was investigated as a function of several parameters, including minor-to-total flow ratios and acceleration nozzle Reynolds number. During these laboratory tests, performance parameters such as concentration enrichment factor (CF), particle losses, collection efficiency curves, cutpoint, and pressure drop were measured. The main objective of these investigations was to optimize the ability of the system to concentrate ambient PM(2.5-10) and PM(1-10) particles. PM(2.5-10) particles were concentrated by a factor of 70 to 150. The flow rate of the concentrated aerosol can range between 12.5 and 50 LPM (L/min). Other features of the system include relatively low-pressure drops in the major and minor flows, low particle losses, and a compact design. Performance evaluation of the system also confirmed that separation and concentration of the PM(2.5-10) particles occurred without any significant distortion of the size distribution, during the concentration process. Similar results were obtained for the PM(1-10) size range. For this size range, concentration enrichment was 70 times, and again, no particle size distribution distortion was observed. The overall performance of this versatile system makes it suitable for inhalation toxicological studies. PMID- 12528043 TI - Effects of inhaled ambient particulate matter on pulmonary antimicrobial immune defense. AB - Respiratory-tract infection, specifically pneumonia, contributes substantially to the increased morbidity and mortality among elderly individuals exposed to airborne particulate matter of <10 microm diameter (PM(10)). These epidemiological findings suggest that PM(10) may act as an immunosuppressive factor that can undermine normal pulmonary antimicrobial defense mechanisms. To investigate whether, and how, compromised pulmonary immunocompetence might contribute to increased mortality, two sets of laboratory studies were performed. The first examined the effects of a single inhalation exposure to concentrated ambient PM(2.5) (CAPS) from New York City air on pulmonary/systemic immunity and on the susceptibility of exposed aged rats to subsequent infection with Streptococcus pneumoniae. The second set of studies determined whether CAPS exposure, at a concentration approximating or somewhat greater than the promulgated 24-h NAAQS of 65 microg/m(3), could exacerbate an ongoing infection. Taken together, results demonstrated that a single exposure of healthy animals to CAPS had little effect on pulmonary immune function or bacterial clearance during subsequent challenge with S. pneumoniae. Alternatively, CAPS exposure of previously infected rats significantly increased bacterial burdens and decreased percentages of lavageable neutrophils and proinflammatory cytokine levels compared to those in infected filtered-air-exposed controls. These studies demonstrate that a single exposure to ambient PM(2.5) compromises a host's ability to handle ongoing pneumococcal infections and support the epidemiological findings of increased pneumonia-related deaths in ambient PM-exposed elderly individuals. PMID- 12528044 TI - Effect of inhaled ultrafine carbon particles on the allergic airway response in ragweed-sensitized dogs. AB - Episodic increases in air pollution have been associated with the exacerbation of asthma symptoms. Ultrafine particles are a component of air pollution and may be involved in causing the adverse health effects associated with high air pollution. We evaluated the effects of ultrafine particle inhalation on immune and airway responses in a beagle dog model of allergic asthma. Six allergic (ragweed sensitive) and six nonallergic dogs were exposed to ultrafine carbon particles (232.3 +/- 2.5 microg/m(3), 35.2 +/- 0.3 nm) for 1 h, followed by a challenge with vehicle (water) as a negative control. Airway resistance was measured during particle exposure and after vehicle challenge. Immune responses 3 days before and after (1 h and 1, 4, 7, and 11 days) particle exposure were assessed by measuring total immunoglobulin E (IgE) and ragweed-specific IgE and IgG in serum and bronchoalveolar lavage fluid (BALF), and cell differentials in BALF. Each dog was exposed a second time to ultrafine carbon particles (251.4 +/- 5.3 microg/m(3), 34.9 +/- 0.5 nm) for 1 h followed by a challenge with ragweed and the same measurements. Airway resistance did not change during particle exposure in any of the dogs, and ragweed-induced airway reactivity was not altered by particle exposure. Total and ragweed-specific serum IgE and total IgE in BALF were higher in allergic dogs at all time points. Particle exposure did not affect antibody levels in serum or BALF in allergic dogs. Nonallergic dogs developed specific IgG in response to multiple inhalation exposures to ragweed, but this was not associated with particle exposure. Neutrophils were elevated in BALF for all groups 1 day after particle exposure. In conclusion, despite the induction of low level inflammation in the lungs of allergic and nonallergic dogs, exposure to ultrafine carbon particles did not alter airway reactivity or immune responses. PMID- 12528045 TI - A comparison of mathematical methods for the determination of in vitro dissolution constants for glass fibers. AB - Biopersistence plays a significant role in determining the potential bioactivity of respirable fibers. In vivo biopersistence in the lung is frequently assessed by in vitro fiber dissolution studies using simulated biological solutions and flow-through techniques. The dissolution rate (k) of a fiber is typically determined by elemental analysis of the flow-through solution to measure the mass of material leached from the fibers over a given time. Various methods may be used to estimate the value of k from these results. The present study compared the in vitro dissolution characteristics of seven experimental glass fiber compositions to those obtained for four recognized fiber compositions (MMVF 10 glass fiber; MMVF 11-glass fiber; MMVF 21-rockwool fiber; crocidolite fiber). Fiber dissolution was examined over a 17-wk period using a flow-through system designed to simulate the conditions encountered by fibers in the extracellular environment of the lung. Mass loss and changes in fiber diameter were determined over time and were then used to calculate k using five different methods. Although the selected methodologies did not produce identical estimations of k for each fiber, the resulting ranking of fiber solubility for each method was consistent. The seven experimental glass fibers were found to have k values intermediate between those of MMVF 11 and MMVF 21. PMID- 12528046 TI - Oxidative damage of sulfur dioxide on various organs of mice: sulfur dioxide is a systemic oxidative damage agent. AB - Cu,Zn-superoxide dismutase (SOD), Se-dependent glutathione peroxidase (GSH-Px), catalase (CAT), and glutathione (GSH) play an important role in attenuating free radical-induced oxidative damage. The purpose of this research was to determine (1) whether sulfur dioxide (SO(2)) increases levels of lipid peroxidation and alters intracellular redox status in multiple organs of mice, and (2) whether SO(2) is a systemic toxic agent. The effect of SO(2) on levels of thiobarbituric acid-reactive substances (TBARS) and GSH and activities of SOD, GSH-Px, and CAT were investigated in nine organs (brain, lung, heart, liver, stomach, intestine, spleen, kidney, and testis) of Kunming albino mice of both sexes. SO(2) at 20 ppm (56 mg/m(3)) was administrated to the animals of SO(2) groups in an exposure chamber for 6 h/day for 7 days while control groups were exposed to filtered air in the same condition. Results show that SO(2) inhalation decreased significantly activities of SOD and GSH-Px in all organs tested in all SO(2) groups, with respect to their corresponding control groups; CAT activities in all organs tested of both sexual mice were significantly unaltered, except CAT activities in livers were significantly lowered by SO(2); SO(2) exposure decreased significantly GSH contents and significantly increased TBARS levels of all organs tested, in comparison with their respective control groups. These results lead to two conclusions: (1) SO(2) is a systemic oxidative damage agent. It results in a significant increase in the lipid peroxidation process in all organs tested of mice of both sexes, which is accompanied by changes of antioxidant status in these organs. (2) SO(2) may cause toxicological damage to multiple organs of animals, and it is suggested that the oxidative damage produced by SO(2) inhalation may influence or promote the progression or occurrence of some disease states of various organs, not only to respiratory system. Further work is required to understand the toxicological role of SO(2) on multiple or even all organs in mammals. PMID- 12528049 TI - Classifying seizures and epilepsy: a synopsis. AB - Seizures can have a number of clinical presentations. Epilepsy, the condition of recurrent unprovoked seizure, can also be categorized in a variety of ways and is associated with several conditions. Seizures and the epilepsies are classified most commonly by a system devised by the International League Against Epilepsy (ILAE) that employs clinical, electroencephalographic, and imaging data. The system has two broad divisions: focal and generalized for seizures and localization related and generalized for the epilepsies. This article surveys all seizure and epilepsy types according to the ILAE scheme. Limitations of the current categorization are discussed, and a recent new proposal for classifying seizures and epilepsy is demonstrated. PMID- 12528050 TI - Antiepileptic medications for the treatment of epilepsy. AB - Epilepsy is one of the most common neurological conditions encountered worldwide. The development of newer antiepileptic drugs (AEDs) has expanded over recent years, and the use of such drugs for indications other than epilepsy has also broadened. These factors insure that the majority of health care providers will be using an increasing number of AEDs in the care of their patients. Contained herein is a review of the mechanism of action of AEDs, individual drugs and their potential drug interactions, and general principles to guide the clinician in selection, implementation, and long-term monitoring of AEDs. PMID- 12528051 TI - Devices in the treatment of epilepsy. AB - The goal of the treatment of epilepsy is to eliminate seizures while causing no side effects. For persons whose seizures are refractory, epilepsy surgery may be an option. In addition, these patients may benefit from the vagus nerve stimulator (VNS), the first device approved for the treatment of refractory epilepsy. Although VNS was the first to be approved, investigators have been interested in the effectiveness of stimulating other brain regions: the cerebellum, thalamus, subthalamic nucleus, and locus coeruleus are a few examples. These studies have produced mixed results. As our understanding of the underlying mechanisms of epilepsy grows, it is likely that we will design better and more effective devices for the treatment of epilepsy. PMID- 12528052 TI - Epilepsy surgery: indications, approaches, and results. AB - The surgical treatment of epilepsy is divided into procedures with curative or palliative goals. Curative procedures are highly effective in rendering the majority of patients seizure free, and palliative procedures often result in marked improvement in seizure frequency, quality of life, or both. This brief overview of epilepsy surgery outlines the goals of surgery, criteria used to determine patient eligibility, the various types of epilepsy surgery, and anticipated outcomes of these approaches. Newer surgical techniques including vagus nerve and deep brain stimulation and gamma knife radiosurgery are also discussed. PMID- 12528053 TI - Neuroimaging of epilepsy. AB - Neuroimaging has an important role in the investigation and treatment of patients with epilepsy. Diagnosis of the underlying substrate in a given patient with epilepsy determines prognosis with higher accuracy than electroencephalography. Neuroimaging techniques include computed tomography (CT) and magnetic resonance imaging (MRI), although CT has a diminished role for diagnosis. MRI is the most appropriate imaging technique in the initial investigation of patients with epilepsy. MRI is the most sensitive technique for the diagnosis of hippocampal sclerosis, tumors, and malformations of cortical development. MRI is also critical for neurosurgical planning. Other imaging techniques such as positron emission tomography (PET) and single photon emission computed tomography are reserved for patients with intractable epilepsy when surgery is contemplated. New developments such as MR spectroscopy, receptor PET, and magnetic source imaging are becoming clinical tools and have the promise of improving diagnosis. PMID- 12528054 TI - Treatment of women with epilepsy. AB - Epilepsy is equally prevalent in men and women. However, for women there are unique concerns related to hormone effects on seizures and the effects of seizures and antiepileptic drugs (AEDs) on reproductive health. Steroid hormones affect neuronal excitability and seizure frequency. Some AEDs reduce the efficacy of oral contraceptive agents, increasing the probability of unplanned pregnancies. AEDs affect bone density. AEDs may alter reproductive hormones resulting in polycystic-appearing ovaries, anovulatory cycles, and infertility. Seizure frequency may change during pregnancy, seizures may cause pregnancy complications, some AEDs are teratogenic, and many cross into breast milk. The treatment of a woman with epilepsy must consider all these issues. PMID- 12528055 TI - Pregnancy in the woman with epilepsy: maternal and fetal outcomes. AB - Pregnancy in women with epilepsy is associated with increased obstetric risks and increased adverse neonatal outcomes. Prior to conception, folic acid should be administered and the antiepileptic drug (AED) regimen should be optimized. Effective control of maternal seizures with the least risk to the fetus is the goal, preferably using AED monotherapy. Periodic monitoring of total and free AED levels is recommended. The "fetal anticonvulsant syndrome" has been described with all of the AEDs and includes major malformations, minor anomalies, microcephaly, cognitive impairment, intrauterine growth retardation, and infant mortality. The most common major malformations are cleft lip/palate, heart defects, and neural tube defects. Prenatal screening should be offered. Supplemental vitamin K(1) should be given to the mother and newborn to prevent neonatal hemorrhagic disorder. Careful planning and management of any pregnancy in women with epilepsy are essential to increase the likelihood of a healthy outcome for the mother and infant. PMID- 12528056 TI - Epilepsy in the elderly. AB - The elderly are the most rapidly growing segment of our population, and onset of epilepsy is higher in this age group than in any other. With advancing age, there are changes in drug absorption, disposition, and binding. In addition, within the elderly are subgroups: (1) elderly healthy except for epilepsy (EH) and (2) elderly with multiple medical problems (EMMP). This review examines the factors that need to be considered when selecting the appropriate antiepileptic drug (AED) for an elderly person with epilepsy. Both the older and newer AEDs are reviewed. PMID- 12528057 TI - Sleep and epilepsy. AB - Quality sleep is essential for patients with epilepsy, but this aspect of their treatment is frequently overlooked. Sleep disturbance can result in daytime drowsiness, worsening memory, and deteriorating seizure control. In a few patients, recognition and treatment of a coexisting sleep disorder can make the difference between complete seizure control and refractory epilepsy. Sleep has effects on interictal epileptiform discharges, with particular implications for diagnostic studies. Sleep disorders can coexist with epilepsy, leading to errors in diagnosis and worsening of seizures. Some specific syndromes demonstrate unique properties related to sleep. Seizures themselves disrupt sleep, even when occurring during the day, with the potential for persistent drowsiness and, perhaps, further memory dysfunction. Finally, most patients with epilepsy are treated with anticonvulsant drugs, some of which can have adverse (or beneficial) effects on sleep. PMID- 12528059 TI - [To our readers: conclusions from inquiries with our readers]. PMID- 12528060 TI - [Do we need newly defined occupational diseases of the respiratory tract?]. PMID- 12528061 TI - [Interstitial pulmonary siderofibrosis: requirements for acceptance as new occupational disease]. AB - BACKGROUND: Pulmonary siderosis is a well established disorder in welders. Internationally more than 150 cases of interstitial pulmonary siderofibrosis are associated with long-standing and heavy exposure to welding fumes at poorly ventilated working places. PATIENTS: Characteristic job histories, lung function analyses and histological examinations as well as elemental microanalysis by energy dispersive X-ray analysis (EDX) are demonstrated from 3 welders with pulmonary siderofibrosis. RESULTS: Histological examinations show a patchy interstitial fibrosis with accumulations of particulate material typical for welding fumes. EDX disclose an increase of iron-load in activated macrophages as well as in lung tissue and a close topographical relationship of welding fume particles and interstitial fibrotic reactions. Lung function analysis showed predominantly loss of pulmonary performance during spiroergometry. CONCLUSIONS: Regarding the actual knowledge about the pathomechanisms of ultrafine particles on lung tissue, the evidence from animal experiments, the histological and electron microscopical results, our own clinical examinations of welders and some epidemiological evidence, we assume a causal relationship of interstitial pulmonary siderofibrosis in welders with long-standing exposure to high concentrations of welding fumes under poor working conditions. PMID- 12528062 TI - [Siderofibrosis, MOTT, and lung transplantation in a worker welding in confined spaces]. PMID- 12528063 TI - [Catheter embolisation into the pulmonary artery. Interventional therapy of a rare form of pulmonary embolism]. AB - Embolisation of a foreign body is a rare cause of pulmonary embolism. We report on a 67 year old female suffering from colorectal cancer. To perform adjuvant chemotherapy a port system was inserted into the right subclavian vein. Routine chest radiography revealed a rupture of the venous port-system, which was embolised into the right pulmonary artery without clinical symptoms. The foreign body was removed without complications using a nitinol goose neck snare via a transfemoral approach and retrieved percutaneously. The reported procedure is safe, decreases operation and hospitalisation time and should therefore be performed in preference to surgical management whenever possible. PMID- 12528064 TI - [Diagnosis and treatment of primary mediastinal liposarcoma]. AB - Mediastinal liposarcoma are of rare entity; there are less than one hundred cases published worldwide. Because of the equivocal and inapparent symptomatology clinical signs often are falsely estimated, and in time of surgery there is generally seen a wide spread tumor growth, so that a radical resection often is impossible. Radiotherapy and chemotherapy are of limited value and mean no chance for curative therapy. The case of a 46 years old patient with an extensive primary liposarcoma of the mediastinum and the results of the review of the literature will be discussed. PMID- 12528065 TI - [Diagnosis, therapy, and prevention of influenza]. PMID- 12528066 TI - [CAPNet--network of competence for community acquired pneumonia: structures and aims]. AB - Community Acquired Pneumonia (CAP) is a disease with high morbidity and mortality in Germany. Apart from the importance of CAP, no reliable data is available about infective germs, resistance of germs, course of the disease, or evidence based treatment. Also basic research should be improved in this field. The German State Ministry for Research (BMBF) has recognised these deficits and supports CAPNet, the network of competence for Community Acquired Pneumonia since 2002. PMID- 12528068 TI - Introduction. New directions in anti-inflammatory therapy and pain management. PMID- 12528069 TI - New directions in symptomatic therapy for patients with osteoarthritis and rheumatoid arthritis. AB - The cyclooxygenase (COX)-2-selective nonsteroidal anti-inflammatory drugs (NSAIDs; coxibs) were developed in order to reduce upper gastrointestinal (GI) side effects associated with traditional nonselective NSAIDs. This article presents an overview of clinical trials showing the efficacy of coxibs for the treatment of patients with arthritis. In osteoarthritis trials, coxibs were more effective than placebo and similarly effective compared with standard doses of traditional NSAIDs. Some studies lasted up to a year and showed effectiveness of coxibs for long-term treatment of patients with osteoarthritis. There are currently few adequately powered trials comparing the efficacy of the 2 first generation coxibs, celecoxib and rofecoxib. Of 2 head-to-head studies comparing the 2 agents, 1 indicated similar efficacy, whereas the other showed the superiority of rofecoxib at a dose of 25 mg daily compared with celecoxib at a dose of 200 mg daily. In studies enrolling patients with rheumatoid arthritis, coxibs also have shown efficacy superior to that of placebo and similar to that of traditional NSAIDs. There are no clinical trials comparing the efficacy of different coxibs for treatment of patients with rheumatoid arthritis. In endoscopic studies, the GI safety and tolerability profile of coxibs has been consistently superior to that of traditional NSAIDs. In large clinical outcome trials, at least 1 of the coxibs, rofecoxib, significantly reduced the risk of confirmed complicated upper GI events compared with the conventional NSAID naproxen. Both rofecoxib and celecoxib reduced the risk of confirmed clinical upper GI events compared with the nonselective NSAIDs naproxen and ibuprofen in patients with osteoarthritis and rheumatoid arthritis not taking low-dose aspirin. Therefore, coxibs provide effective relief of pain from osteoarthritis and rheumatoid arthritis, with efficacy that is comparable to traditional NSAIDs, but with a significantly lower incidence of GI complications. PMID- 12528070 TI - Coxibs: Evolving role in pain management. AB - Traditional pain management strategies have relied on the use of opioids, nonsteroidal anti-inflammatory drugs (NSAIDs), and acetaminophen, as well as other adjuvant analgesics. However, the limited activity of these drugs and the substantial adverse effects associated with their use has left many patients without dependable options for effective treatment. Recent advances in the understanding of pain and its pathophysiologic mechanisms have led to the development of novel therapeutic options. Cyclooxygenase (COX)-2-specific inhibitors (coxibs) have an established efficacy in the treatment of chronic arthritic pain comparable to that of traditional NSAIDs, without the degree of gastrointestinal (GI) complications commonly attributed to NSAID use. Recent trials also have shown the effectiveness of 1 of the coxibs for relief of chronic lower back pain. Numerous studies have shown that coxibs are efficacious for the management of acute pain in various clinical settings, including orthopedic surgery, dental surgery, and dysmenorrhea. The superior safety profile of coxibs in conjunction with a comparable efficacy to nonselective NSAIDs supports the use of coxibs in balanced analgesic regimens. Decreased GI and antiplatelet effects of coxibs compared to traditional NSAIDs provide the potential to incorporate coxibs into the pain management algorithm used to treat cancer pain. PMID- 12528071 TI - The gastrointestinal effects of nonselective NSAIDs and COX-2-selective inhibitors. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs), which include aspirin, are among the most frequently prescribed medications worldwide. The main factor limiting use of NSAIDs is concern about their gastrointestinal (GI) side effects. The purpose of this article is to review the incidence, pathophysiology, and risk factors of GI side effects associated with NSAID therapy. Upper GI symptoms, such as dyspepsia, occur in 15% to 60% of NSAID users, twice as often as in individuals not taking NSAIDs. The prevalence of gastric or duodenal ulcers in patients taking NSAIDs regularly is approximately 15% to 30%. The annual incidence of NSAID-related clinical upper GI events (complicated and symptomatic ulcers) is approximately 2.5% to 4.5%, with the annual incidence of serious complications (severe bleeding, perforation, and obstruction) about 1% to 1.5%. A history of ulcer or GI complications, advanced age, concomitant anticoagulation therapy or corticosteroid use, and high-dose or multiple NSAID therapy are associated with an increased risk of GI events during NSAID therapy. The cyclooxygenase (COX)-2 specific inhibitors (coxibs) have been developed in order to improve the GI safety and tolerability profile of therapy with NSAIDs. In numerous clinical trials, coxibs have been shown to have efficacy similar to that of nonselective NSAIDs, but are associated with significantly fewer endoscopic ulcers. In addition, 2 large outcome trials indicated that coxibs can also reduce the incidence of clinically important GI events. PMID- 12528072 TI - Anti-inflammatory agents and renal function. AB - The purpose of this review is to provide an overview of the effect of traditional nonsteroidal anti-inflammatory drugs (NSAIDs) and cyclooxygenase (COX)-specific-2 inhibitors (coxibs) on renal function. Both COX isoforms (COX-1 and COX-2) constitutively are expressed in the adult mammalian kidney and contribute to the biosynthesis of prostaglandins (PGs). Inhibition of COX activity in the kidney by NSAIDs has relatively mild consequences in healthy individuals, but can lead to serious adverse events in patients whose renal function is PG dependent. Most studies have reported transient decreases in sodium excretion upon initiation of therapy with either traditional NSAIDs or coxibs. In patients whose renal function is dependent on prostanoids, both nonselective NSAIDs and coxibs can affect the glomerular filtration rate. Changes in renal function may result in hypertension and edema. Several studies have compared the effects of traditional NSAIDs, celecoxib, and rofecoxib on blood pressure and incidence of edema, but the results have not been consistent, as the trials used different clinical models and study designs. No trials to date have been performed with rigorous study designs that allow meaningful drug comparisons at comparably effective doses. In consequence, patients who are at risk for adverse renal events should be monitored with the same caution when receiving coxibs as when receiving treatment with nonselective NSAIDs. They include patients with congestive heart failure or renal or hepatic disease, as well as those of advanced age receiving therapy with diuretics or angiotensin-converting enzyme (ACE) inhibitors. PMID- 12528073 TI - Challenges of managing chronic pain in the elderly. AB - Effective management of chronic pain in the elderly has posed a formidable challenge to the medical community. Practicing physicians are confronted with the complexity of controlling chronic pain in a patient population that often presents with multiple health problems and side effects from long-term use of pain medications. This study presents an overview of commonly encountered barriers to effective management of pain in the elderly and suggests strategies to optimize treatment in this patient population. A high incidence of comorbidities in the elderly frequently necessitates the use of multiple therapies, the combination of which potentially increases the toxicity experienced by this population, which is already particularly susceptible to medication side effects. When determining optimal strategy for managing chronic pain in the elderly, particular attention needs to be paid to past medical history and use of over-the-counter medications. In addition, careful monitoring of age-related changes in physiologic functions is needed, as they may impact drug plasma concentrations and increase susceptibility to adverse events. When traditional pharmacologic agents (such as acetaminophen and nonselective nonsteroidal anti-inflammatory drugs) are deemed inappropriate in high-risk patients because of ineffectiveness or side effects, physicians must consider the use of alternative drugs, such as coxibs, which have comparable efficacy to traditional analgesics, with a superior side effects profile. PMID- 12528076 TI - Grading autonomic dysfunction in chronic fatigue syndrome. PMID- 12528077 TI - Henoch Schonlein purpura in children and adults: is it one entity? PMID- 12528078 TI - Hemodynamics instability score in chronic fatigue syndrome and in non-chronic fatigue syndrome. AB - OBJECTIVE: In studying patients with chronic fatigue syndrome (CFS) we developed a method that confers numerical expression to the degree of blood pressure and heart rate lability, ie, the 'hemodynamic instability score' (HIS). The HIS in CFS patients differed significantly from healthy subjects. The present investigation compares the HIS in CFS, non-CFS chronic fatigue and patients with recurrent syncope. METHODS: Patients with CFS (n = 21), non-CFS chronic fatigue (n = 24), syncope of unknown cause (n = 44), and their age and sex-matched healthy controls (n = 21) were evaluated with a standardized head-up tilt test (HUTT). Abnormal reactions (endpoints) on HUTT were classified 'clinical outcomes' (cardioinhibitory or vasodepressor reaction, orthostatic hypotension, postural tachycardia syndrome) and 'HIS endpoint', i.e. HIS >-0.98. RESULTS: The highest incidence of endpoints was noted in patients with CFS (79%), followed by patients with syncope of unknown cause (46%), non-CFS chronic fatigue (35%), and healthy subjects (14%). Presyncope or syncope during tilt occurred in 38% of CFS patients, 21% of patients with non-CFS chronic fatigue, and 43% of patients with recurrent syncope. The average HIS values were: CFS = +2.02 (SD 4.07), non-CFS chronic fatigue = -2.89 (SD 3.64), syncope = -3.2 (SD 3.0), healthy = -2.48 (4.07). The odds ratios for CFS patients to have HIS >-0.98 was 8.8 compared with non-CFS chronic fatigue patients, 14.6 compared with recurrent syncope patients, and 34.8 compared with healthy subjects. CONCLUSION: The cardiovascular reactivity in patients with CFS has certain features in common with the reactivity in patients with recurrent syncope or non-CFS chronic fatigue, such as the frequent occurrence of vasodepressor reaction, cardioinhibitory reaction, and postural tachycardia syndrome. Apart from to these shared responses, the large majority of CFS patients exhibit a particular abnormality which is characterized by HIS values >-0.98. Thus, HIS >-0.98 lends objective criteria to the assessment of CFS. PMID- 12528079 TI - Henoch-Schonlein purpura in children and adults: clinical differences in a defined population. AB - OBJECTIVE: To examine epidemiologic, clinical, and outcome differences between children and adults with Henoch-Schonlein purpura (HSP) in a well-defined population. PATIENTS AND METHODS: Retrospective study of unselected patients with HSP seen at the only referral hospital for the Lugo region of Northwest Spain between 1980 and 2000. Patients were classified according to the criteria proposed by Michel et al. Two well-differentiated age groups were established for comparison: children (under 14 years of age) and adults (over 20 years of age). Also, to assess possible differences in the outcome, only those patients with at least 1 year of follow-up were included in the study. RESULTS: Seventy-three children and 31 adults fulfilled the inclusion criteria described above. Unlike in children, HSP in adults was more common in males. While in children, HSP manifested more commonly in fall and winter, summer and winter were the most common seasons of onset in adults. The frequency of gastrointestinal manifestations was similar in both groups. However, during the course of the disease, 6 of the 31 adults (19%) had severe renal manifestations and another 4 (13%) renal insufficiency. In children, by contrast, the frequency of severe renal manifestations or renal insufficiency during the course of the disease was significantly reduced compared with adults. After 6 years' median follow-up in children, complete recovery was observed in most cases. However, after 5 years' median follow-up, almost 40% of adults had persistent hematuria and 3 of them (10%) renal insufficiency that required hemodialysis in 2 cases. CONCLUSIONS: HSP is generally benign and self-limited in children and more severe in adults. PMID- 12528080 TI - Interferon-induced Raynaud's syndrome. AB - OBJECTIVE: To review the clinical features, diagnosis, treatment, and outcome of interferon-induced Raynaud's phenomenon. METHODS: The medical literature was reviewed from 1967 to November 2001 with the assistance of a MEDLINE search using the key words: Raynaud, Interferon, ischemia, thrombosis and necrosis. A qualitative review was performed after the articles were abstracted and the relevant information was summarized. RESULTS: Twenty-four cases of interferon induced Raynaud's phenomenon (including our patient) are described. Interpheron alpha was the most common causative agent (14 cases). The symptoms appeared weeks to years after beginning treatment and varied from mild vasospasm to occlusion of digital arteries and tissue necrosis (14 cases), sometimes necessitating finger amputation (6 patients). Digital plethysmography, arteriography and capillaroscopy were valuable diagnostic tools. In 4 cases, cardiac, ophthalmic, or central nervous system drug-induced ischemia accompanied the peripheral Raynaud's phenomenon. Of the 15 cases with a documented outcome, withdrawal of the drug alone resulted in complete (6 patients) or partial (1 patient) recovery. In the others, supportive therapy was needed. The recovery period lasted from 2 weeks to 3 months. In 2 patients, continuation of treatment was possible. CONCLUSIONS: Raynaud's phenomenon and related complications must be recognized as possible side effects of interferon therapy. Early diagnosis and withdrawal of the drug may prevent unnecessary morbidity and disability. PMID- 12528081 TI - Autoimmune disease complicating antiviral therapy for hepatitis C virus infection. AB - OBJECTIVE: To review autoimmune disease complicating therapy with type I interferons (IFNs), specifically in the setting of hepatitis C virus (HCV) infection. METHODS: This study describes 13 reported cases of drug-induced systemic lupus erythematosus (SLE) associated with IFN therapy for the period reported during 1990-2002 by searching MEDLINE. In addition, 2 additional patients are presented, 1 with SLE and 1 with an antineutrophil cytoplasmic antibody (ANCA)-positive nephritis, with long-term follow-up. RESULTS: Of 13 cases of SLE-like syndromes caused by IFN, 2 occurred in patients being treated for HCV infection. Two occurred in patients with rheumatoid arthritis (RA); 1 had Sjogren's syndrome (SS), and 1 laryngeal papillomatosis. The rest were receiving IFN for hematologic malignancies. Symptoms developed between 2 weeks and 7 years after initiation of therapy. Most developed fever and arthralgias/arthritis. Other findings included serositis manifested by tachycardia, dyspnea and pleural effusions, headaches, and hair loss. All had a positive antinuclear antibody (ANA), and the majority had double stranded (ds) DNA antibodies. Two additional patients with chronic HCV infection developed autoimmune disease after combination treatment with IFN-alpha and ribavirin. In each patient, autoimmune disease manifested as severe joint pains, myalgias, fever, rash, and proteinuria. Skin and renal biopsy specimens showed vasculitis and crescentic glomerulonephritis (GN) in the first case, and typical histologic findings of lupus nephritis in the second; clinical and laboratory features were consistent with Wegener's granulomatosis and SLE, respectively. Although both patients had mixed polyclonal cryoglobulins, they were HCV RNA and HCVAb negative. Both received corticosteroids, with gradual clinical and biochemical improvement and without recurrence of viremia. CONCLUSIONS: Autoimmune disorders occur in 4% to 19% of patients receiving IFN-alpha, though SLE-like syndromes are only seen in 0.15% to 0.7%. Clinical and laboratory features of SLE in this setting resemble idiopathic disease, with a generally good outcome after discontinuance of the drug. RELEVANCE: Type I IFNs may cause autoimmune disease such as SLE. As the armamentarium of drugs expands to include other biologics, such as the tumor necrosis factor (TNF)-alpha-inhibiting drugs, the development of autoimmune diseases induced by these drugs is an important consideration for diagnosis and appropriate treatment. Semin Arthritis Rheum 32:163-173. PMID- 12528082 TI - Importance of planning ovulation induction therapy in systemic lupus erythematosus and antiphospholipid syndrome: a single center retrospective study of 21 cases and 114 cycles. AB - OBJECTIVE: To analyze the results and complications of ovulation induction therapy (OIT) in women with systemic lupus erythematosus (SLE) and/or the antiphospholipid syndrome (APS). METHODS: A retrospective study of 21 women followed in a single tertiary-referral French center who underwent 114 OIT cycles with or without in vitro fertilization and embryo transfer (IVFET). RESULTS: Before OIT, SLE was present in 6 women, APS in 3, SLE-related APS in 3, and discoid lupus in 1. Eight women had no identified disease and underwent 36 cycles of OIT. Diagnosis (SLE, n = 3; primary APS, n = 5) was made after OIT complication: spontaneous abortion (n = 5), SLE flare (n = 2), and thrombophlebitis (n = 1). Five women with known disease intentionally concealed their history from their gynecologists and underwent 34 cycles. Forty-four cycles were planned in 11 women, in 3 of them after complications of prior OIT performed without particular therapy and monitoring. Eighteen pregnancies occurred, which ended in 9 live births, 4 fetal deaths, and 5 embryonic losses. The pregnancy rate was higher with gonadotropin and/or gonadotropin-releasing hormone analog (GnRHa) (25% of cycles) than with clomiphene (4% of cycles, P <.0001). When the gynecologists did not know the underlying disease, three-quarters of pregnancies induced by OIT with IVFET ended in embryonic losses or fetal deaths. In contrast, 6 of 7 pregnancies induced by planned OIT with IVFET ended in live births (P <.0001). Phlebothromboses were observed only with gonadotropin treatment. The SLE flare rate was higher with gonadotropin and/or GnRHa (27% of cycle) than with clomiphene (6%, NS). It also was higher (30%) when the gynecologists did not know the underlying disease than in the planned procedures (10%, NS). CONCLUSIONS: The OIT may precipitate SLE or APS. A careful review of the patient's history and appropriate laboratory tests should be undertaken before OIT. Clomiphene complications are rare. When gonadotropins are prescribed, preventive anti inflammatory therapy should be considered in women with SLE, in addition to heparin and/or anti-aggregant therapy in patients with asymptomatic anti phospholipid antibodies or prior thrombotic events. PMID- 12528083 TI - Sternoclavicular infectious arthritis in previously healthy adults. AB - OBJECTIVE: To define characteristics of sternoclavicular infection (SCI) in previously healthy patients. METHODS: SCI in a previously healthy man is reported along with 4 similar cases found by surveying the hospital's database; 22 previously reported cases were culled from the literature and summarized. RESULTS: The frequency of SCI in healthy adults was 0.5% of all bone and joint infections admitted to the hospital. The clinical and bacteriologic features were similar to previous reports in nonselected SCI patients. Computerized tomography (CT) scan, arthrocenthesis, and biopsy were required for diagnosis. Complications included abscess formation, mediastinitis, and sepsis. The majority of patients were treated by surgical drainage and antibiotics. The final outcome was good, without mortality or long-term morbidity. CONCLUSIONS: Although SCI is a rare infection in healthy adults, it should be considered in the differential diagnosis of a painful sternoclavicular joint. Prompt diagnosis and appropriate treatment of SCI results in excellent outcome in most cases. PMID- 12528084 TI - Vocational rehabilitation in patients with chronic rheumatic diseases: a systematic literature review. AB - OBJECTIVE: To describe the effectiveness of vocational rehabilitation programs for patients with chronic rheumatic diseases by means of a systematic review of the literature. METHODS: Data were obtained by a computer-aided and manual search of the literature from 1980 until May 2001. Vocational rehabilitation programs had to be clearly defined interventions specifically aimed at having patients with rheumatic diseases reenter or remain in the work force. The vocational rehabilitation programs had to be executed by one or more health professionals. Outcome of the intervention had to be described in terms of vocational status (work disability, sick leave, job modification, paid occupation, retraining). RESULTS: Six studies were identified. All were uncontrolled studies. Follow-up periods ranged 2-84 months. Five of six vocational rehabilitation programs consisted of multidisciplinary intervention and 15% to 69% of the patients successfully returned to work. CONCLUSIONS: Although 5 of 6 studies showed a marked positive effect of vocational rehabilitation on work status, proof of the benefit of these interventions is limited, mainly due to methodologic differences and shortcomings. RELEVANCE: Work disability is a major consequence of the disease in patients with rheumatic conditions. More and more attention is being paid to preventing disability and promoting return to work. Knowledge regarding the effectiveness of vocational rehabilitation programs is insufficient. Semin Arthritis Rheum 32:196-203. PMID- 12528085 TI - Ileal pouchitis-related arthritis. PMID- 12528086 TI - What is the significance of a journal's impact factor? PMID- 12528087 TI - Tumor necrosis factor-alpha blockade as a therapeutic strategy in heart failure (RENEWAL and ATTACH): unsuccessful, to be specific. PMID- 12528088 TI - Effect of selective and nonselective beta-blockers on resting energy production rate and total body substrate utilization in chronic heart failure. AB - BACKGROUND: In chronic heart failure (CHF) beta-blockers reduce myocardial oxygen consumption and improve myocardial efficiency by shifting myocardial substrate utilization from increased free fatty acid oxidation to increased glucose oxidation. The effect of selective and nonselective beta-blockers on total body resting energy production rate (EPR) and substrate utilization is not known. METHODS: Twenty-six noncachectic patients with moderately severe heart failure (New York Heart Association class II or III, left ventricular ejection fraction < 0.40) were treated with carvedilol (37.5 +/- 13.5 mg/12 h) or bisoprolol (5.4 +/- 3.0 mg/d) for 6 months. Indirect calorimetry was performed before and after 6 months of treatment. RESULTS: Resting EPR was decreased in carvedilol (5.021 +/- 0.803 to 4.552 +/- 0.615 kJ/min, P <.001) and bisoprolol group (5.230 +/- 0.828 to 4.978 +/- 0.640 kJ/min, P <.05; nonsignificant difference between groups). Lipid oxidation rate decreased in carvedilol and remained unchanged in bisoprolol group (2.4 +/- 1.4 to 1.5 +/- 0.9 mg m(2)/kg min versus 2.7 +/- 1.1 to 2.5 +/- 1.1 mg m(2)/kg min, P <.05). Glucose oxidation rate was increased only in carvedilol (2.6 +/- 1.4 to 4.4 +/- 1.6 mg m(2)/kg min, P <.05), but did not change in bisoprolol group. CONCLUSIONS: Both selective and nonselective beta blockers reduce total body resting EPR in noncachectic CHF patients. Carvedilol compared to bisoprolol shifts total body substrate utilization from lipid to glucose oxidation. PMID- 12528089 TI - beta-blockers in heart failure--much more than heart rate reduction. PMID- 12528091 TI - Race, ethnicity, and heart failure. PMID- 12528090 TI - Racial differences in treatment-seeking delays among heart failure patients. AB - BACKGROUND: Treatment-seeking delays for heart failure (HF) symptoms are significantly high. However, earlier studies did not closely examine race as a characteristic that could potentially influence delay times. The purpose of this study was (1) to describe racial differences in treatment-seeking delays for HF symptoms and (2) to identify racial differences in hospital readmission rates, functional status, and total length of stay. METHODS AND RESULTS: A retrospective chart review of all patients admitted with HF at a Veterans Administration facility was conducted. The study sample consisted of 753 patients: 456 Caucasians (60.6%), 220 African Americans (29.2%), 41 Asians (5.4%), and 36 Hispanics (4.8%). The average prehospital delay time was 2.9 +/- 0.7 days. Mean delay times were significantly longer for African Americans than for Caucasians, Asians, and Hispanics (P =.019). African Americans also had significantly higher readmission rates (P =.001) and lower functional status (higher New York Heart Association functional class) (P =.034). There were no significant racial differences in total length of stay for HF admissions. CONCLUSION: The current study supports that racial differences exist in treatment-seeking behaviors for HF symptoms, hospital readmission rates, and functional status. A better understanding of treatment-seeking behaviors of HF patients with different racial characteristics may be key to early recognition and prevention of complications in this high-risk population; it may be beneficial in identifying patients at risk for treatment delays and potentially poorer outcomes. PMID- 12528092 TI - Plasma matrix metalloproteinase and inhibitor profiles in patients with heart failure. AB - BACKGROUND: Changes in myocardial matrix metalloproteinases (MMPs) and the inhibitors of MMPs (TIMPs) have been demonstrated in congestive heart failure (CHF). The first objective of this study was to measure plasma profiles of MMPs and TIMPs in CHF patients (n = 24; 62 +/- 3 years; left ventricular ejection fraction [LVEF] = 24 +/- 2%) and age-matched nonfailing patients (n = 48; 63 +/- 2 years; LVEF >/= 55%). Cytokines such as tumor necrosis factor (TNF)-alpha can induce MMP expression in vitro. The second objective of this study was to determine the relationship between soluble TNF-alpha receptors (TNFR1; TNFR2) and MMP plasma profiles. METHODS AND RESULTS: Plasma levels of MMP-2, MMP-9, MMP-8, TIMP-1, TIMP-2, TNF-alpha, TNFR1, and TNFR2 were measured by enzyme-linked immunosorbent assay kits. Plasma MMP-9 levels were increased in CHF patients (25 +/- 6 versus 72 +/- 15 ng/mL, P <.05). Interestingly, plasma levels of MMP-8 were decreased in CHF patients (16 +/- 2 versus 9 +/- 2 ng/mL, P <.05). The MMP-9/TIMP 1 ratio was increased by 3-fold, whereas the MMP-9/TIMP-2 ratio was increased by 16-fold in CHF patients (both P <.05). With a 48-week follow-up in CHF patients, an absolute reduction in plasma TNFR1 from baseline was accompanied by reduced MMP-9 levels (-30 +/- 16 ng/mL; P =.058), whereas stable or increased plasma TNFR1 resulted in persistently elevated MMP-9 levels. CONCLUSIONS: The unique findings of this study were 2-fold. First, a discordant change in plasma MMP and TIMP levels occurred in CHF patients. Second, changes in cytokine activity were related to changes in plasma MMP levels. These changes in MMP/TIMP levels likely reflect the progression and/or acceleration of the LV remodeling process in CHF. Thus serial measurements of plasma MMP/TIMP levels may hold diagnostic/prognostic significance in CHF patients. PMID- 12528093 TI - Acupuncture inhibits sympathetic activation during mental stress in advanced heart failure patients. AB - BACKGROUND: In heart failure (HF) patients, muscle sympathetic nerve activity is increased, and HF patients with the greatest sympathetic activation have the poorest prognosis. In animals, acupuncture is sympathoinhibitory, and the most profound sympathoinhibition occurs in animals with the highest resting sympathetic nerve activity. The purpose of this study was to test the hypothesis that acupuncture is sympathoinhibitory in humans with HF. METHODS AND RESULTS: Fifteen advanced HF patients underwent acute mental stress testing before and during (1) "real" acupuncture (n = 10), (2) non-acupoint acupuncture (n = 10), and (3) no-needle acupuncture control (n = 10). Muscle sympathetic nerve activity (MSNA) was recorded using peroneal microneurography. Resting MSNA was not different before and after acupuncture (52 +/- 22 versus 50 +/- 21 bursts/min, P = NS). During mental stress, SNA increased significantly. This increase was eliminated following real acupuncture (mean delta MSNA pre-acupuncture versus post-acupuncture: 149 +/- 171 versus -169 +/- 130, P =.03), but not after non acupoint or no-needle acupuncture controls. The changes in blood pressure and heart rate during mental stress were not attenuated by real or control acupuncture. CONCLUSION: Acute acupuncture attenuates sympathoexcitation during mental stress in advanced HF patients. PMID- 12528094 TI - Exaggerated cardiovascular effects of cocaine in conscious dogs with pacing induced dilated cardiomyopathy. AB - BACKGROUND: The aim of this study was to explore the characteristics and mechanisms of the cardiovascular effects of cocaine in dilated cardiomyopathy. METHODS AND RESULTS: We studied the cardiovascular responses to acute intravenous cocaine (1 mg/kg) in 8 conscious, chronically instrumented dogs before and after the development of dilated cardiomyopathy induced by rapid ventricular pacing. To help elucidate the role of altered baroreflex function in mediating the cardiovascular effects of cocaine, we also studied responses in 3 conscious, chronically instrumented dogs that had undergone surgical sinoaortic baroreceptor denervation. Cocaine produced greater increases in heart rate (+57 +/- 8% from 112 +/- 5 beats/min versus +28 +/- 3% from 100 +/- 4 beats/min; P <.01), first derivative of left ventricular pressure (+30 +/- 5% from 1,714 +/- 147 mm Hg/sec versus +15 +/- 3% from 3,032 +/- 199 mm Hg/sec; P <.01), coronary vascular resistance (+28 +/- 5% from 2.3 +/- 0.3 mm Hg/mL/min versus +11 +/- 5% from 2.2 +/- 0.3 mm Hg/mL/min; P <.05) and plasma norepinephrine concentration (+130 +/- 31% from 462 +/- 102 pg/mL versus +86 +/- 32% from 286 +/- 77 pg/mL; P <.05) in dogs with dilated cardiomyopathy as compared to controls. In addition, responses were much more rapid in onset following the development of dilated cardiomyopathy. Chronotropic and inotropic responses to cocaine were similarly rapid and exaggerated in dogs after baroreceptor denervation. CONCLUSIONS: Cocaine produces rapid and exaggerated chronotropic, inotropic, and coronary vasoconstrictor responses in conscious dogs with pacing-induced dilated cardiomyopathy. Alterations in arterial baroreflex function may play a role in these observations, which in turn may underlie the clinically observed association between cocaine and heart failure. PMID- 12528095 TI - Ranolazine, a partial fatty acid oxidation (pFOX) inhibitor, improves left ventricular function in dogs with chronic heart failure. AB - BACKGROUND: Abnormalities of energy metabolism are often cited as key elements in the progressive worsening of left ventricular (LV) dysfunction that characterizes the heart failure (HF) state. The present study tested the hypothesis that partial inhibition of fatty acids will ameliorate the hemodynamic abnormalities associated with HF. METHODS AND RESULTS: Chronic HF (LV ejection fraction 27 +/- 1%) was produced in 13 dogs by intracoronary microembolizations. Hemodynamic and angiographic measurements were made before and 40 minutes after intravenous administration of ranolazine, a partial fatty acid oxidation (pFOX) inhibitor. Ranolazine was administered as an intravenous bolus dose of 0.5 mg/kg followed by a continuous infusion for 40 minutes at a constant rate of 1.0 mg / kg / hr. Ranolazine significantly increased LV ejection fraction (27 +/- 1 versus 36 +/- 2, P =.0001), peak LV +dP/dt (1712 +/- 122 versus 1900 +/- 112 mm Hg/sec, P =.001), and stroke volume (20 +/- 1 versus 26 +/- 1 mL). These improvements occurred in the absence of any effects on heart rate or systemic pressure. In 8 normal healthy dogs, ranolazine had no effect on LV ejection fraction or any other index of LV function. CONCLUSIONS: In dogs with HF, acute intravenous administration of the pFOX inhibitor ranolazine improves LV systolic function. The absence of any hemodynamic effects of ranolazine in normal dogs suggests that the drug is devoid of any positive inotropic effects and acts primarily by optimizing cardiac metabolism in the setting of chronic HF. PMID- 12528096 TI - Disparate force-frequency effects of pimobendan and dobutamine in conscious dogs with tachycardia-induced cardiomyopathy. AB - BACKGROUND: This study was designed to examine how a calcium sensitizer, pimobendan, affected a force-frequency response (FFR) as compared to the beta adrenergic agonist dobutamine. METHODS AND RESULTS: Left ventricular (LV) contractility and relaxation were evaluated by the slope (Ees) of the LV end systolic pressure-volume relation and the time constant (Tau) of LV pressure decay. Using 6 conscious dogs with tachycardia-induced heart failure, the FFR was examined before and after administration of dobutamine (6 microg/kg/min) or pimobendan (0.5 mg/kg). Despite the similar inotropic and lusitropic action at the baseline heart rate, pimobendan and dobutamine showed different FFR and relaxation-frequency responses. Before administration of these drugs, there was no significant increase in LV contractility and relaxation by increasing heart rate. However, dobutamine amplified FFR (Ees: +3.1 +/- 1.4, P <.05) as compared with Ees for a comparable increase in heart rate before administration of the drug. On the other hand, pimobendan showed relatively mild amplification of FFR compared with dobutamine (Ees: +1.9 +/- 1.1, P <.05). The relaxation-frequency response tended to increase with dobutamine but not with pimobendan. CONCLUSIONS: Mild amplification of FFR observed in pimobendan suggests that this agent could be used more safely than beta-adrenergic agent when heart rate is increased, as seen with exercise. PMID- 12528097 TI - The benefits of early treatment in rheumatoid arthritis: confounding by indication, and the issue of timing. PMID- 12528098 TI - Disposal of dying cells: a balancing act between infection and autoimmunity. PMID- 12528099 TI - Selective cyclooxygenase 2 inhibitors, aspirin, and cardiovascular disease: a reappraisal. PMID- 12528100 TI - Radiographic data from recent randomized controlled trials in rheumatoid arthritis: what have we learned? PMID- 12528101 TI - Adalimumab, a fully human anti-tumor necrosis factor alpha monoclonal antibody, for the treatment of rheumatoid arthritis in patients taking concomitant methotrexate: the ARMADA trial. AB - OBJECTIVE: To evaluate the efficacy and safety of adalimumab (D2E7), a fully human monoclonal tumor necrosis factor alpha antibody, in combination with methotrexate (MTX) in patients with active rheumatoid arthritis (RA) despite treatment with MTX. METHODS: In a 24-week, randomized, double-blind, placebo controlled study, 271 patients with active RA were randomly assigned to receive injections of adalimumab (20 mg, 40 mg, or 80 mg subcutaneously) or placebo every other week while continuing to take their long-term stable dosage of MTX. The primary efficacy end point was the American College of Rheumatology criteria for 20% improvement (ACR20) at 24 weeks. RESULTS: An ACR20 response at week 24 was achieved by a significantly greater proportion of patients in the 20-mg, 40-mg, and 80-mg adalimumab plus MTX groups (47.8%, 67.2%, and 65.8%, respectively) than in the placebo plus MTX group (14.5%) (P < 0.001). ACR50 response rates with the 20-mg, 40-mg, and 80-mg adalimumab dosages (31.9%, 55.2%, and 42.5%, respectively) were significantly greater than that with placebo (8.1%) (P = 0.003, P < 0.001, and P < 0.001, respectively). The 40-mg and 80-mg doses of adalimumab were associated with an ACR70 response (26.9% and 19.2%, respectively) that was statistically significantly greater than that with placebo (4.8%) (P < 0.001 and P = 0.020). Responses were rapid, with the greatest proportion of adalimumab-treated patients achieving an ACR20 response at the first scheduled visit (week 1). Adalimumab was safe and well tolerated; comparable numbers of adalimumab-treated patients and placebo-treated patients reported adverse events. CONCLUSION: The addition of adalimumab at a dosage of 20 mg, 40 mg, or 80 mg administered subcutaneously every other week to long-term MTX therapy in patients with active RA provided significant, rapid, and sustained improvement in disease activity over 24 weeks compared with MTX plus placebo. PMID- 12528102 TI - Influence of disease-modifying therapy on radiographic outcome in inflammatory polyarthritis at five years: results from a large observational inception study. AB - OBJECTIVE: To determine the effect of early treatment with disease-modifying antirheumatic drugs (DMARDs) in reducing radiographic progression over a 5-year period in patients with new-onset inflammatory polyarthritis. METHODS: Three hundred thirty-five consecutive patients with paired radiographs obtained 1 year and 5 years after enrollment in a population-based arthritis register were studied. Logistic regression was used to model differences in baseline factors associated with the start of DMARDs. The time from symptom onset to first use of DMARDs was stratified to represent 4 groups: no DMARD use, <6 months, 6-12 months, and >12 months. Radiographs of the hands and feet were scored using the Larsen method. Progression in the Larsen score was evaluated as a 5-year score adjusted for the first film score. Negative binomial regression was used to compare Larsen score progression for each of the 3 treatment groups with that for patients not receiving DMARDs. Results were then adjusted for severity, based on propensity modeling. RESULTS: Patients who received treatment had more radiographic progression than did patients who were untreated. Coefficients (95% confidence intervals), expressed as a multiple of the Larsen score in DMARD treated patients compared with untreated patients, were as follows: 1.6 (1.1-2.3) for <6 months, 2.4 (1.5-3.6) for 6-12 months, and 2.0 (1.4-2.8) for >12 months. As expected, patients receiving treatment had more severe disease at baseline. Using the propensity score as a method of adjusting for disease severity, the influence of treatment on outcome became attenuated as follows: 1.1 (0.8-1.7) for <6 months, 1.6 (1.0-2.6) for 6-12 months, and 1.5 (1.0-2.2) for >12 months. This effect was also seen in the crude Larsen score at year 5. CONCLUSION: In this observational study, DMARD treatment was a marker not only of worse disease at presentation but also of the radiographic state and radiographic progression at 5 years. After adjustments were made for baseline disease severity, earlier therapy was shown to have a beneficial effect on outcome. PMID- 12528103 TI - Survival in rheumatoid arthritis: a population-based analysis of trends over 40 years. AB - OBJECTIVE: To evaluate trends in and risk factors for mortality among patients with rheumatoid arthritis (RA) over a 40-year period. METHODS: A population-based inception cohort was assembled from among all Rochester, Minnesota residents ages > or =18 years who were first diagnosed with RA (fulfilling the 1987 American College of Rheumatology criteria for RA) between January 1, 1955 and December 31, 1994. Patients were followed up longitudinally through their entire medical records (including all inpatient and outpatient care by any provider) until death or migration from the county. Survival was described using the Kaplan-Meier method. Observed and expected survival were compared using the log-rank test, and standardized mortality ratios (SMRs) with expected survival were based on the sex and age of the study population and death rates from the Minnesota life tables. Cox proportional hazards models were used to estimate the influence of extraarticular manifestations and comorbidities, controlling for age, sex, body mass index (BMI), smoking, and rheumatoid factor positivity. RESULTS: Survival in this RA cohort was significantly lower than that expected in the population (P < 0.001) over the entire time period. Patients with RA were at significantly higher risk of death, with an SMR of 1.27 (95% confidence interval 1.13-1.41). Excess mortality among women was more pronounced than among men, with SMRs of 1.41 and 1.08, respectively. Presence of > or =1 extraarticular manifestation was the strongest predictor of mortality after adjusting for age, sex, BMI, smoking, and rheumatoid factor positivity. CONCLUSION: Survival in RA patients is significantly lower than expected. The strongest predictors of survival appear to be those related to RA disease complications, specifically, extraarticular manifestations of the disease and comorbidities. More attention should be paid to mortality as an outcome measure in RA. PMID- 12528104 TI - Functional disability in rheumatoid arthritis patients compared with a community population in Finland. AB - OBJECTIVE: To compare Health Assessment Questionnaire (HAQ) scores of patients with rheumatoid arthritis (RA) with HAQ scores from a sex- and age-adjusted population. METHODS: Patients with RA (n = 1,095) and control subjects (n = 1,530) completed a mailed questionnaire that comprised the HAQ, pain and global health scores, education level, and comorbidities, as well as height, weight, and lifestyle attitudes, including smoking and exercise habits. RESULTS: The HAQ scores increased (indicating declining function) with older age in patients and controls. The HAQ scores were above the reference values (>95th percentile of the HAQ scores of the age- and sex-matched population) in 17-45% of women with RA and in 7-32% of men with RA ages 30-79 years, while the HAQ scores of the patients > or =80 years were similar to those of the age- and sex-matched population. In a logistic regression model, the odds ratio for disability (HAQ score > or =1; at least some difficulties in most activities of daily living) was 7.7 (95% confidence interval 5.3-11.1; P < 0.001) among patients with RA compared with community controls, when adjusted for age, sex, education, smoking, exercise, body mass index, number of comorbidities, and pain. CONCLUSION: RA is associated with a >7-fold risk of disability compared with that in a general population of adults in the same community. The impact of disability due to RA appears to be greater in younger and middle-age people than in elderly patients. PMID- 12528105 TI - Elucidation of the relationship between synovitis and bone damage: a randomized magnetic resonance imaging study of individual joints in patients with early rheumatoid arthritis. AB - OBJECTIVE: To simultaneously image bone and synovium in the individual joints characteristically involved in early rheumatoid arthritis (RA). METHODS: Forty patients with early, untreated RA underwent gadolinium-enhanced magnetic resonance imaging (MRI) of the second through fifth metacarpophalangeal joints of the dominant hand at presentation, 3 months, and 12 months. In the first phase (0 3 months), patients were randomized to receive either methotrexate alone (MTX) or MTX and intraarticular corticosteroids (MTX + IAST) into all joints with clinically active RA. The MTX-alone group received no further corticosteroids until the second phase (3-12 months), when both groups received standard therapy. RESULTS: In the first phase, MTX + IAST reduced synovitis scores more than MTX alone. There were significantly fewer joints with new erosions on MRI in the former group compared with the latter. During the second phase, the synovitis scores were equivalent and a similar number of joints in each group showed new erosions on MRI. In both phases, there was a close correlation between the degree of synovitis and the number of new erosions, with the area under the curve for MRI synovitis the only significant predictor of bone damage progression. In individual joints, there was a threshold effect on new bone damage related to the level of synovitis; no erosions occurred in joints without synovitis. CONCLUSION: In early RA, synovitis appears to be the primary abnormality, and bone damage occurs in proportion to the level of synovitis but not in its absence. In the treatment of patients with RA, outcome measures and therapies should focus on synovitis. PMID- 12528106 TI - Screening for atherosclerosis in patients with rheumatoid arthritis: comparison of two in vivo tests of vascular function. AB - OBJECTIVE: Inflammation appears to play a central role in atherosclerosis, and endothelial damage mediated by systemic inflammation may contribute to the increased cardiovascular mortality in rheumatoid arthritis (RA). Brachial artery flow-mediated dilatation (FMD) and pulse wave analysis (PWA) are measures of vascular function. The aim of this study was to determine if FMD and PWA are abnormal in patients with RA. METHODS: Twenty-five RA patients and 25 matched healthy controls were studied. All were free of traditional cardiovascular risk factors. FMD was measured in all subjects. PWA was performed in 18 RA patients and 18 controls, with results expressed as large and small artery compliance (C1 and C2). Modified Sharp scores were calculated in 13 RA patients. RESULTS: Results (mean +/- SD) in RA patients and controls, respectively, were as follows: FMD 107.6 +/- 4.6% versus 108.5 +/- 4.1% (P = 0.49), C1 14.8 +/- 2.8 ml/mm Hg x 10 versus 17.9 +/- 3.1 ml/mm Hg x 10 (P = 0.0033), C2 4.5 +/- 2.3 ml/mm Hg x 100 versus 7.7 +/- 3.7 ml/mm Hg x 100 (P = 0.0039). There was an inverse correlation between C2 and modified Sharp scores in the RA patients (Spearman's rho -0.69, P = 0.0085). CONCLUSION: FMD was normal in these RA patients, whereas arterial compliance was markedly reduced. PWA appears to be a more sensitive measure of vascular dysfunction than FMD in RA and may be the preferred surrogate marker of vascular dysfunction in longitudinal studies of RA patients. The inverse correlation between C2 and the modified Sharp score, a measure that reflects disease activity over time, supports the notion that chronic inflammation plays a role in RA-associated atherosclerosis. PMID- 12528107 TI - Reduced arterial elasticity in rheumatoid arthritis and the relationship to vascular disease risk factors and inflammation. AB - OBJECTIVE: Rheumatoid arthritis (RA) is associated with increased rates of cardiovascular disease. Reduced small artery elasticity (SAE) and large artery elasticity (LAE) and increased systemic vascular resistance (SVR) have been found in other high-risk groups. In the present study, we sought to determine whether arterial elasticity was reduced and SVR was increased in RA patients compared with controls matched for coronary artery disease (CAD) status, and to relate the results to vascular disease risk factors, including measures of inflammation. METHODS: Arterial elasticity was assessed by pulse wave analysis in RA patients with (n = 15) and without (n = 38) CAD, and in controls matched 1:1 for age, sex, and CAD status. Vascular risk factors, including high-sensitivity C-reactive protein (hsCRP), soluble vascular cell adhesion molecule 1 (sVCAM-1), and serum amyloid A (SAA) levels, were assessed. RESULTS: SAE and LAE were significantly lower and SVR was significantly higher in RA patients than in controls. RA patients also had higher levels of hsCRP, SAA, and sVCAM-1. SAE and LAE values were inversely correlated with markers of inflammation. Associations of SAE and LAE with RA were independent of conventional risk factors, but were dependent on markers of inflammation. CONCLUSION: Vascular function is abnormal in RA, with reduced SAE and LAE and increased SVR relative to controls. Arterial elasticity is inversely associated with measures of inflammation. These measures may be clinically useful in the detection and monitoring of vascular disease in RA. PMID- 12528108 TI - The effect of HLA-DR on susceptibility to rheumatoid arthritis is influenced by the associated lymphotoxin alpha-tumor necrosis factor haplotype. AB - OBJECTIVE: HLA-DRB1, a major genetic determinant of susceptibility to rheumatoid arthritis (RA), is located within 1,000 kb of the gene encoding tumor necrosis factor (TNF). Because certain HLA-DRB1*04 subtypes increase susceptibility to RA, investigation of the role of the TNF gene is complicated by linkage disequilibrium (LD) between TNF and DRB1 alleles. By adequately controlling for this LD, we aimed to investigate the presence of additional major histocompatibility complex (MHC) susceptibility genes. METHODS: We identified 274 HLA-DRB1*04-positive cases of RA and 271 HLA-DRB1*04-positive population controls. Each subject was typed for 6 single-nucleotide polymorphisms within a 4.5-kb region encompassing TNF and lymphotoxin alpha (LTA). LTA-TNF haplotypes in these unrelated individuals were determined using a combination of family data and the PHASE software program. RESULTS: Significant differences in LTA-TNF haplotype frequencies were observed between different subtypes of HLA-DRB1*04. The LTA-TNF haplotypes observed were very restricted, with only 4 haplotypes constituting 81% of all haplotypes present. Among individuals carrying DRB1*0401, the LTA-TNF 2 haplotype was significantly underrepresented in cases compared with controls (odds ratio 0.5 [95% confidence interval 0.3-0.8], P = 0.007), while in those with DRB1*0404, the opposite effect was observed (P = 0.007). CONCLUSION: These findings suggest that the MHC contains genetic elements outside the LTA-TNF region that modify the effect of HLA-DRB1 on susceptibility to RA. PMID- 12528109 TI - Laser-mediated microdissection facilitates analysis of area-specific gene expression in rheumatoid synovium. AB - OBJECTIVE: Current approaches to analyzing gene expression in rheumatoid arthritis (RA) synovium are based on RNA isolated either from cultured synovial cells or from synovial biopsy specimens. This strategy does not, in general, allow distinction of specific gene expression between cells originating from different synovial areas, due to potential mixture of expression profiles. Therefore, we established the combination of laser-mediated microdissection (LMM) and differential display to analyze profiles of gene expression in histologically defined areas of rheumatoid synovium. The present study was undertaken to establish parameters for this technique and assess its usefulness for gene expression analysis. METHODS: Cryosections derived from RA synovial tissues were used to obtain cell samples from synovial lining versus sublining, using a microbeam laser microscope. RNA was isolated and analyzed by nested RNA arbitrarily primed-polymerase chain reaction (RAP-PCR) for differential display fingerprinting. Differentially expressed bands were cut out, and PCR products were eluted, cloned, and sequenced. Differential expression of identified sequences was confirmed by in situ hybridization and immunohistochemistry analysis. RESULTS: Microdissected sections of RA synovial tissue containing approximately 600 cells yielded enough RNA to produce a reproducible RNA fingerprint pattern. Several genes could be identified as being expressed differentially between the synovial lining and the sublining, and their expression could be confirmed at the messenger RNA and protein levels. CONCLUSION: The combination of LMM and RAP-PCR presents a valuable tool to obtain novel insights into the area-dependent differential regulation of gene expression in RA synovium. Both known and previously unknown genes were revealed with this technique. This study is the first to demonstrate the potential of this analytic strategy in the investigation of a nonmalignant, multifactorial, inflammatory disease. PMID- 12528110 TI - Control of fibroblast-like synoviocyte proliferation by macrophage migration inhibitory factor. AB - OBJECTIVE: The hyperplasia of fibroblast-like synoviocytes (FLS) is considered essential to the evolution of joint destruction in rheumatoid arthritis (RA), but the mechanisms underlying FLS proliferation remain poorly understood. Macrophage migration inhibitory factor (MIF) is a cytokine that has recently been shown to exert proinflammatory effects on RA FLS. This study sought to identify the mechanisms of activation of FLS by MIF, and to assess the effects of MIF on synovial cell proliferation. METHODS: Human RA FLS were treated with recombinant MIF, interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha (TNF alpha), and/or anti-MIF monoclonal antibodies (mAb). Proliferation was measured with tritiated thymidine incorporation. Nuclear factor kappa B (NF-kappa B) and mitogen-activated protein (MAP) kinase activation were measured with immunohistochemistry and Western blotting, respectively. RESULTS: FLS proliferation was significantly increased by MIF. IL-1 beta and TNFalpha also induced proliferation, but these effects were prevented by neutralization with anti-MIF mAb. Activation of NF-kappa B was induced by IL-1 beta, but not by MIF. Anti-MIF mAb had no effect on IL-1 beta-induced NF-kappa B nuclear translocation. By contrast, MIF induced phosphorylation of extracellular signal-regulated kinase (ERK) MAP kinase. ERK antagonism, but not NF-kappa B antagonism, prevented the effect of MIF on FLS proliferation. CONCLUSION: These data suggest that MIF may regulate RA synovial hyperplasia by acting directly and via involvement in the effects of IL-1 beta and TNFalpha. In addition, the effects of MIF on FLS activation are independent of NF-kappa B, and dependent on ERK MAP kinase. These data suggest an important therapeutic potential for MIF antagonism in RA. PMID- 12528111 TI - Integrin-mediated adhesion of human articular chondrocytes to cartilage. AB - OBJECTIVE: To determine 1) the kinetics and strength of adhesion of human articular chondrocytes to a cut cartilage surface, and 2) the role of specific integrins in mediating such adhesion, using an in vitro model. METHODS: Human articular chondrocytes isolated from cadaveric donors (mean +/- SD age 38 +/- 13 years) were cultured in high-density or low-density monolayer. Following release from culture with trypsin and a 2-2.5-hour recovery period, chondrocytes were analyzed either for adhesion to cartilage or for integrin expression by flow cytometry. RESULTS: Following culture in monolayer, adhesion of chondrocytes to cartilage increased with time, from 6-16% at 10 minutes to a maximum of 59-82% at 80-320 minutes. After 80 minutes of adhesion, the resistance of cells to flow induced shear stress (50% detachment) was approximately 21 Pa. Chondrocyte adhesion to cartilage decreased with pretreatment of cells with monoclonal antibodies that bound to and blocked certain integrins. After an 80-minute incubation time, adhesion of chondrocytes cultured in high-density monolayer decreased from the value of IgG1-treated controls (55%) with blocking of the beta1 integrin subunit (to 23%) or with blocking of alpha 5 beta 1 (to 36%). Following expansion of chondrocytes in low-density monolayer, the mechanisms of adhesion to cartilage were generally similar. After an 80-minute incubation time, adhesion of chondrocytes cultured in low-density monolayer decreased from the value of IgG1-treated controls (62%) with blocking of the beta1 integrin subunit (to 30%) or with blocking of alpha 5 beta 1 (to 44%). Additionally, adhesion of these cells decreased to 46% by blocking of alpha v beta 5, with a similar trend in effect for chondrocytes cultured in high-density monolayer. Blocking of the alpha 1 or alpha 3 integrin subunits or alpha v beta 3 had no detectable effect on adhesion, even though these receptors were detected by flow cytometry. CONCLUSION: Under the culture and seeding conditions studied, beta1, alpha 5 beta 1, and alpha v beta 5 integrins mediate human chondrocyte adhesion to cartilage. These chondrocyte integrins have a potential role in the initial adhesion and retention of chondrocytes at a cartilage defect site following clinical procedures of chondrocyte transplantation. PMID- 12528112 TI - Induction of aggrecanase 1 (ADAM-TS4) by interleukin-1 occurs through activation of constitutively produced protein. AB - OBJECTIVE: To study the production of aggrecanase 1 (ADAM-TS4) in monolayer chondrocytes, capsular fibroblasts, and cartilage. METHODS: Bovine nasal and articular cartilage, monolayer chondrocytes, and capsular fibroblasts were incubated in the absence and presence of interleukin-1 (IL-1). ADAM-TS4 production was evaluated by immunofluorescence or by Western blot analysis. Aggrecanase activity was measured in cells grown on an immobilized peptide substrate, and peptide cleavage was monitored by enzyme-linked immunosorbent assay. RESULTS: There was constitutive production of ADAM-TS4 in both cells and tissue. The protein was associated with the extracellular matrix based on the observation that the staining could be reduced following treatment of chondrocytes with heparin or exposure to chondroitinase ABC. Interestingly, there was no detectable change in the abundance of ADAM-TS4 in response to IL-1. Western blot analysis of cell lysates from IL-1-stimulated chondrocytes showed no evidence of increased ADAM-TS4 production, but resulted in activation of ADAM TS4. The activation was associated with an increased generation in the aggrecanase neoepitope NITEGE in nasal cartilage in response to IL-1. These data suggest that induction of aggrecanase activity both in cells and in cartilage by IL-1 may involve the stimulation of an activator of ADAM-TS4. Consistent with this observation, culture of chondrocytes on a solid support containing a peptide substrate resulted in the generation of aggrecanase-mediated cleavage that could be blocked by selective inhibitors of ADAM-TS4. CONCLUSION: These data support the hypothesis that ADAM-TS4 is constitutively produced in these cells and tissue, and that stimulation by IL-1 results in aggrecanase activation. Thus, the activator could be a potential target by which to control aggrecanase-mediated degradation in arthritic diseases. PMID- 12528113 TI - Induction of matrix metalloproteinase 1 gene expression is regulated by inflammation-responsive transcription factor SAF-1 in osteoarthritis. AB - OBJECTIVE: To identify specific transcription factors that are activated in the cartilage tissue of osteoarthritis (OA) patients and are involved in the expression of matrix metalloproteinase 1 (MMP-1). METHODS: DNase I footprint and electrophoretic mobility shift assays were performed to identify active elements of the MMP1 promoter and the transcription factors that interact with it. The relative abundance of the involved transcription factor in the cartilage was determined by immunohistochemical analysis. The role of serum amyloid A activating factor 1 (SAF-1) in MMP-1 expression was assessed by transient transfection assay with plasmids containing either a functional or an antisense SAF-1 complementary DNA. RESULTS: A novel promoter element was detected in the human MMP1 gene, and the inflammation-responsive transcription factor SAF-1 was found to interact with it. SAF-1 activity was detected in the chondrocytes of OA cartilage, where most of the cells stained positive for SAF-1. Overexpression of SAF-1 in OA chondrocytes increased the expression of the MMP1 promoter, and interference of endogenous SAF-1 activity by antisense SAF-1 messenger RNA inhibited interleukin-1-mediated MMP1 promoter activity. CONCLUSION: The results indicate that SAF-1 is involved in the regulation of MMP1 gene expression and it is highly abundant in the articular cartilage chondrocytes of OA patients. The data also demonstrate that control of SAF-1 activity can suppress induced expression of MMP-1. Conceivably, SAF-1 could be a potential therapeutic target to control the overexpression of MMP-1 associated with the pathogenesis of OA. PMID- 12528114 TI - Tachykinin expression in cartilage and function in human articular chondrocyte mechanotransduction. AB - OBJECTIVE: To assess whether substance P and the corresponding neurokinin 1 (NK1) receptor are expressed in human articular cartilage, and whether these molecules have a role in chondrocyte mechanotransduction. METHODS: Transgenic studies, immunohistochemistry, Western blotting, and reverse transcriptase-polymerase chain reaction were used to assess the expression of the preprotachykinin (PPT) gene, substance P, and NK1 in developing mice, in adult human articular cartilage, and in human chondrocytes in culture. Chondrocytes obtained from PPT knockout mice and human articular chondrocytes were mechanically stimulated in the presence or absence of inhibitors of substance P signaling, and cell membrane potentials or relative levels of aggrecan messenger RNA (mRNA) were measured. RESULTS: Replacing a region of the PPT gene transcriptional site that contains a dominant repressor of the proximal promoter activity with the constitutive minimal promoter of the human beta-globin promoter allowed expression of a marker gene in areas of chondrogenesis during mouse development and in adult chondrocytes grown in culture. Adult human articular chondrocytes expressed endogenous PPT mRNA, substance P, and the corresponding NK1 receptor in vivo and in vitro. Blockade of substance P signaling by a chemical antagonist to the NK1 receptor inhibited chondrocyte responses to mechanical stimulation. CONCLUSION: Substance P is expressed in human articular cartilage and is involved in chondrocyte mechanotransduction via the NK1 receptor in an autocrine and paracrine manner. This suggests that substance P and the NK1 receptor have roles in the maintenance of articular cartilage structure and function that were previously unrecognized. PMID- 12528115 TI - Jaccoud's arthropathy in systemic lupus erythematosus: differentiation of deforming and erosive patterns by magnetic resonance imaging. AB - OBJECTIVE: To evaluate alterations of the soft tissues, tendons, and bones as detected by magnetic resonance imaging (MRI) in patients with systemic lupus erythematosus (SLE)-associated arthritis of the finger joints. METHODS: Both hands of 14 patients with SLE of various activities and durations and with arthritis and/or deformities of the finger joints were examined by MRI and conventional radiography. Coronal T1-weighted spin-echo (with and without gadolinium contrast), axial T2-weighted turbo spin-echo, coronal fat-suppressed short tau inversion recovery, and 3-dimensional double-echo steady-state sequences were acquired and analyzed, and the findings were compared with those from conventional radiographs. RESULTS: MRI detected periarticular capsular swelling in all 14 patients, joint effusion in 7, edematous tenosynovitis in 6, proliferative tenosynovitis (flexor and/or extensor tendons) in 4, and intraarticular signs of synovial membrane hypertrophy in 10 patients, 9 of whom showed enhancement after administration of contrast medium. Bony erosions were identified in 8 patients by MRI; conventional radiography missed these erosions in 2 of the 8 patients. Four of the 14 patients were designated as Jaccoud's arthropathy index-positive, and all 4 showed severe edematous tenosynovitis and capsular swelling, but no signs of bony erosions despite longstanding disease (mean 21.5 years). Four of the 10 patients with mild deformity exhibited prominent soft tissue pathology, with minimal destruction of bone; the other 6 patients had bony alterations that resembled rheumatoid arthritis. CONCLUSION: In SLE patients with arthritis of the finger joints, MRI detects characteristic signs of soft tissue pathology (e.g., capsular swelling, edematous and proliferative tenosynovitis, synovial hypertrophy) and bony alterations (e.g., erosions, some of which are missed by conventional radiography). MRI thus helps to distinguish different types of lupus arthritis/Jaccoud's arthropathy, which allows more differentiated treatment strategies and monitoring. PMID- 12528116 TI - High-dose cyclophosphamide without stem cell transplantation in systemic lupus erythematosus. AB - OBJECTIVE: High-dose chemotherapy followed by hematopoietic stem cell transplantation is increasingly being studied as a treatment for severe autoimmune disorders, such as systemic lupus erythematosus (SLE). High-dose cyclophosphamide, the foundation of virtually all conditioning regimens for stem cell transplantation, is not myeloablative; therefore, when high-dose cyclophosphamide is used alone, autografting, with its potential for reinfusing autoreactive effector cells, is not required. We undertook this study to investigate the safety and efficacy of high-dose cyclophosphamide without stem cell transplantation in refractory SLE. METHODS: We treated 14 patients with moderate-to-severe SLE that had been refractory to corticosteroids and one or more additional immunosuppressive drugs. All patients received 50 mg/kg of cyclophosphamide for 4 consecutive days followed by 5 microg/kg granulocyte colony-stimulating factor until the neutrophil count was 1 x 10(9)/liter for 2 consecutive days. Patients were followed up monthly for disease activity using the physician's global assessment, SLE Disease Activity Index, and assessment of functioning of involved organs. The Responder Index for Lupus Erythematosus was used to define partial or complete response. RESULTS: The median time to achieve a neutrophil count of 0.5 x 10(9)/liter was 14 days (range 11-22 days) after the last dose of cyclophosphamide. Patients required a median of 2 units (range 2-5) of packed red blood cells, and a median of 16 days (range 0-23 days) elapsed from the last dose of cyclophosphamide to the last platelet transfusion. There were no deaths or fungal infections. Significant improvements in physician's global assessment (mean difference 1.4; P < 0.0001), SLE Disease Activity Index (mean difference 4.1; P = 0.0039), and prednisone dosage (mean difference 14.9 mg/day; P = 0.002) were observed. Responses, including 5 durable complete responses, were observed in all organ systems (renal, central nervous system, and skin) with involvement that had led to patient enrollment. CONCLUSION: High-dose cyclophosphamide without stem cell transplantation leads to rapid hematopoietic reconstitution and has significant clinical benefit in patients with refractory SLE. Therefore, this approach deserves further study. PMID- 12528117 TI - Increased salivary gland tissue expression of Fas, Fas ligand, cytotoxic T lymphocyte-associated antigen 4, and programmed cell death 1 in primary Sjogren's syndrome. AB - OBJECTIVE: To assess salivary gland tissues obtained from patients with primary Sjogren's syndrome (SS) for the gene expression profile of the candidate genes TNFRSF6 (Fas), TNFSF6 (FasL), SSA1 (Ro52 alpha and the splice variant Ro52 beta), SSB (La), CTLA4, PDCD1 (PD-1), and ORM2, which were selected on the basis of their putative participation in salivary gland inflammation. METHODS: Quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was used to examine the expression of messenger RNA (mRNA). Tissue localization of the expressed proteins was detected by immunohistochemistry. RESULTS: Expression of mRNA was increased for Fas (5.1-fold; P < 0.001), FasL (8.8-fold; P < 0.05), cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) (11.2-fold; P < 0.01), programmed cell death 1 (PD-1) (15.2-fold; P < 0.05), Ro52 alpha (3.0 fold; P < 0.01), La (2.3-fold; P < 0.05), and orosomucoid 2 (ORM2) (4.4-fold; P < 0.05) in patients compared with controls when GAPDH was used as endogenous standard in duplex runs. In single runs using 2 other endogenous standards (18S and beta-actin), statistically significant differences between patients and controls were confirmed for expression of Fas, FasL, CTLA-4, and PD-1, but this difference was not observed for Ro52 alpha, La, and ORM2. Expression of Ro52 beta mRNA was similar in patients and controls. CONCLUSION: The present study demonstrates a substantial increase in expression of the negative regulator molecules PD-1 and CTLA-4 and the apoptotic signal molecules Fas and FasL in patients with primary SS compared with controls, which corresponded to the immunomorphologic pattern. The results strongly indicate that these molecules have central roles in the inflammatory process in the salivary glands of patients with primary SS. PMID- 12528118 TI - Association of IL1A gene polymorphisms with susceptibility to and severity of systemic sclerosis in the Japanese population. AB - OBJECTIVE: To analyze the frequencies of haplotypes of single-nucleotide polymorphisms (SNPs) of the IL1A gene (at -889, +4729, and +4845) in patients with systemic sclerosis (SSc) and in healthy control subjects, and to determine whether the IL1A gene haplotype is associated with SSc susceptibility or disease severity. METHODS: We studied 60 patients with SSc (34 with diffuse cutaneous SSc and 26 with limited cutaneous SSc) and 70 healthy control subjects. Polymorphisms of the IL1A gene were genotyped by direct sequencing using the ABI Prism 377 Sequence Detection System. The LDSupport program, which was recently developed in our laboratory, was used to estimate the haplotype frequencies of SNPs in the study population. RESULTS: We confirmed the presence of 2 SNPs at positions -889 (C/T) and +4845 (G/T) of the IL1A gene, as previously reported. We also identified a novel SNP at position +4729 (T/C). Six haplotypes, CTG (49.7%), TCT (14.7%), CCT (20.3%), TTG (13.2%), CCG (1.4%), and TTT (0.7%), were found in the healthy controls. In contrast, only 2 haplotypes, CTG (95%) and TCT (5%), were detected in the SSc patients. Notably, the CTG haplotype was present at a significantly higher frequency in the SSc patients than in the healthy controls (P < 0.0001). We also examined the relationship between the CTG/CTG diplotype frequencies and interstitial lung disease (ILD), a major complication of SSc, as an indicator of disease severity. All SSc patients with ILD had the CTG/CTG diplotype, whereas the frequency of this diplotype was only 67% in patients without ILD. CONCLUSION: Our observations suggest that the CTG haplotype of the IL1A gene may be an important marker for the susceptibility to, and the severity of, SSc. PMID- 12528119 TI - Identification of an IL1A gene segment that determines aberrant constitutive expression of interleukin-1 alpha in systemic sclerosis. AB - OBJECTIVE: To further explore the mechanisms of transcriptional regulation of the IL1A gene in fibroblasts from the affected skin of patients with systemic sclerosis (SSc). METHODS: The transcription activity of IL1A was estimated by nuclear run-on assay. A reporter construct (pLuc.-1437) was prepared that contained a 1.5-kb genomic fragment extending from -1437 to +39 relative to the transcription start site of the IL1A gene inserted upstream of the luciferase gene. The DNA protein binding activity for the cis-element of the IL1A gene was determined by electrophoretic mobility shift assay (EMSA). RESULTS: Affected fibroblasts exhibited a strong nuclear run-on signal, indicative of constitutive transcription activity of the IL1A gene. Luciferase assay indicated high levels of transcription activity of pLuc.-1437 in transiently transfected fibroblasts from affected skin; this was absent in transiently transfected control fibroblasts. Analysis of constructs containing a series of 5' deletions of pLuc. 1437 revealed that the sequence between -96 and -82 (nuclear factor-interleukin-1 alpha [NF-IL-1 alpha]) was the most important for transcription of the IL1A gene in affected SSc fibroblasts. NF-IL-1 alpha contains a 5'-GGAA-3' sequence that is essential for specific binding of members of the Ets nuclear protein family. Using EMSA, a DNA protein complex was detected in samples containing nuclear extracts from affected fibroblasts and the NF-IL-1 alpha probe, whereas no specific band was detected in samples from normal fibroblasts. The DNA protein complex disappeared with the use of NF-IL-1 alpha probes that had mutations of the Ets binding sites. CONCLUSION: These findings suggest that a DNA binding protein containing the Ets domain is constitutively expressed only in fibroblasts from the affected skin of SSc patients and regulates the transcription of the IL1A gene, which contributes to the fibrogenic phenotype of fibroblasts in SSc. PMID- 12528120 TI - A comparison between anti-Th/To- and anticentromere antibody-positive systemic sclerosis patients with limited cutaneous involvement. AB - OBJECTIVE: To compare anti-Th/To-positive and anticentromere antibody (ACA) positive patients with limited cutaneous systemic sclerosis (lcSSc). METHODS: We reviewed the medical records of 107 anti-Th/To-positive patients and 365 ACA positive patients who were first evaluated during 1985-2000. ACA was detected by indirect immunofluorescence on HEp-2 cell substrate, and anti-Th/To was detected by RNA immunoprecipitation with K562 cell extracts. Patients were included if they had a clinical diagnosis of lcSSc, and excluded if they had >1 SSc associated serum autoantibody. RESULTS: The final study groups comprised 87 lcSSc patients with anti-Th/To antibodies and 306 with ACAs. Anti-Th/To-positive patients were younger (P < 0.04) and had a shorter disease duration at their first evaluation (P < 0.003). Patients with anti-Th/To antibodies had more subtle skin changes, less severe vascular involvement, and less frequent distal esophageal hypomotility. Both groups had a higher frequency of "intrinsic" pulmonary hypertension than has been previously reported in the literature (28% and 19% of anti-Th/To-positive and ACA-positive patients, respectively), perhaps due to referral bias. Patients in the anti-Th/To group more often had radiographic evidence of interstitial lung disease (48% versus 13% of the ACA group; P < 0.0001). Scleroderma renal crisis was uncommon (4 cases), but occurred exclusively in the anti-Th/To group. Survival among the anti-Th/To-positive patients was reduced compared with that in the ACA group (P < 0.02). CONCLUSION: Patients with anti-Th/To and those with ACA most often develop lcSSc and have a high frequency of "intrinsic" pulmonary hypertension. Compared with the ACA patients, anti-Th/To lcSSc patients have more subtle cutaneous, vascular, and gastrointestinal involvement, but more often have certain features typically seen in diffuse scleroderma, such as pulmonary fibrosis and scleroderma renal crisis, as well as reduced survival. PMID- 12528121 TI - Human scleroderma sera contain autoantibodies to protein components specific to the U3 small nucleolar RNP complex. AB - OBJECTIVE: To determine whether antifibrillarin autoantibodies in scleroderma patients are associated with autoantibodies to protein components specific for U3 small nucleolar RNP (U3 snoRNP). METHODS: Sera from 220 scleroderma patients were examined for antinucleolar autoantibodies (ANoA) and for antibodies to fibrillarin and the U3 snoRNP-specific proteins Mpp10 and hU3-55K. Clinical correlates were determined for the different autoantibody specificities. RESULTS: Fifty-nine of the 220 patients were positive for ANoA, and 31 of these patients were antifibrillarin positive. Anti-hU3-55K was found in 10 patients, all of whom were antifibrillarin positive. Twenty-nine patients had anti-Mpp10 antibodies; 23 of these were antifibrillarin positive and 6 were antifibrillarin negative. ANoA, including antifibrillarin, anti-hU3-55K, and anti-Mpp10, were associated with diffuse, rather than limited, systemic or localized scleroderma. Esophageal and lung involvement were more common in patients with antifibrillarin and anti-Mpp10 antibodies, and the highest frequency was in patients with anti-Mpp10 alone. CONCLUSION: Antifibrillarin autoantibodies are associated with autoantibodies to protein components specific to U3 snoRNP, particularly Mpp10. The prevalence of anti-Mpp10 antibodies in antifibrillarin-positive patients suggests that the U3 snoRNP particle is a source of immunogenic/antigenic material for the anti-snoRNP response in scleroderma. Autoantibodies to snoRNP components were more frequent in patients with diffuse scleroderma than in those with either the limited systemic or localized forms. The increased expression of these antibodies in patients with the more severe form of scleroderma, coupled with the observations that fibrillarin expression is positively linked to collagen expression in fibroblasts and that fibrillarin is overexpressed in scleroderma fibroblasts, suggests a source of snoRNP to initiate and maintain these autoantibody responses. PMID- 12528122 TI - Long-term efficacy and safety of etanercept in children with polyarticular-course juvenile rheumatoid arthritis: interim results from an ongoing multicenter, open label, extended-treatment trial. AB - OBJECTIVE: To evaluate the long-term efficacy and safety of etanercept in children with juvenile rheumatoid arthritis (JRA) participating in an ongoing multicenter, open-label, extended-treatment trial. All patients had been participants in an initial randomized efficacy and safety trial of etanercept. METHODS: Etanercept was administered at a dosage of 0.4 mg/kg (maximum 25 mg) subcutaneously twice each week. Safety and efficacy evaluations were performed every 3-4 months. The JRA 30% definition of improvement (DOI) was defined as improvement of > or =30% in at least 3 of 6 response variables used to assess disease activity, with no more than 1 variable worsening by more than 30%. RESULTS: At the time of analysis, 48 of the 58 patients (83%) were still enrolled in the study; 43 of them (74%) had completed 2 years of treatment. Of these 43 patients, 81% met the JRA 30% DOI, 79% met the JRA 50% DOI, and 67% met the JRA 70% DOI. Ten children started low-dose methotrexate after year 1. Of the 32 children taking prednisone, the dosage was decreased to <5 mg/day in 26 (81%). Two children had serious infections (varicella with aseptic meningitis in one and complicated sepsis in the other). In general, adverse events were of the types seen in a general pediatric patient population. CONCLUSION: Children with severe, longstanding, methotrexate-resistant polyarticular JRA demonstrated sustained clinical improvement with >2 years of continuous etanercept treatment. Etanercept was generally well-tolerated. There were no increases in the rates of adverse events over time. However, children taking etanercept should be monitored closely for infections. PMID- 12528123 TI - Arthropathy in juvenile hemochromatosis. AB - OBJECTIVE: To evaluate whether arthropathy is associated with juvenile hemochromatosis and, if so, to assess the relationship between this feature and other clinical features of the disease. METHODS: Clinical, laboratory, and radiologic evidence of arthropathy was studied in 8 Greek patients with genetically proven juvenile hemochromatosis. Osteopenia and osteoporosis were assessed by bone mineral density measurement. RESULTS: Seven of the 8 patients had articular manifestations. The main affected joint was the metacarpophalangeal joint, and the arthritis was progressive independent of phlebotomy therapy. Osteopenia was observed in 2 patients, and osteoporosis in 2 patients. CONCLUSION: Arthropathy may be present in patients with juvenile hemochromatosis, with features similar to those found in patients with hemochromatosis type 1. PMID- 12528124 TI - Rapid induction of peroxisome proliferator-activated receptor gamma expression in human monocytes by monosodium urate monohydrate crystals. AB - OBJECTIVE: Peroxisome proliferator-activated receptor gamma (PPAR gamma) is a member of the nuclear hormone receptor superfamily and functions as a key regulator of lipid and glucose metabolism, atherosclerosis, and inflammatory responses. This study was undertaken to evaluate the biologic role of PPAR gamma in self-limiting episodes of acute gouty arthritis. To do this, we investigated PPAR gamma expression by monosodium urate monohydrate (MSU) crystal-stimulated monocytes, and we studied the effects of PPAR gamma ligands on crystal-induced acute inflammation. METHODS: PPAR gamma expression by MSU crystal-stimulated human peripheral blood mononuclear cells was determined by reverse transcription polymerase chain reaction and immunostaining. Expression of CD36 on monocytes was detected by flow cytometric analysis. The effects of PPAR gamma ligands on in vitro crystal-induced cytokine production and on in vivo cellular infiltration during crystal-induced acute inflammation were also investigated. RESULTS: MSU crystals rapidly and selectively induced PPAR gamma expression by monocytes. Gene expression was detected as early as 2 hours, and maximum expression was observed at 4 hours after stimulation. The induced PPAR gamma was functional, since a PPAR gamma ligand was able to up-regulate CD36 expression on monocytes. A natural ligand of PPAR gamma, 15-deoxy-Delta(12,14)-prostaglandin J(2) (15deoxy-PGJ(2)), significantly reduced the crystal-induced production of cytokines by monocytes. Indomethacin inhibited cytokine production only at high concentrations, and an antidiabetic thiazolidinedione (troglitazone) failed to exert significant effects. Administration of troglitazone and 15deoxy-PGJ(2) significantly prevented cellular accumulation in a mouse air-pouch model of MSU crystal-induced acute inflammation. CONCLUSION: Rapid induction of PPAR gamma expression on monocytes by MSU crystals may contribute, at least in part, to the spontaneous resolution of acute attacks of gout. PMID- 12528125 TI - Adenosine A2A or A3 receptors are required for inhibition of inflammation by methotrexate and its analog MX-68. AB - OBJECTIVE: Low-dose weekly methotrexate therapy remains a mainstay in the treatment of inflammatory arthritis. Results of previous studies demonstrated that adenosine, acting at one or more of its receptors, mediates the antiinflammatory effects of methotrexate in animal models of both acute and chronic inflammation. We therefore sought to establish which receptor(s) is involved in the modulation of acute inflammation by methotrexate and its nonpolyglutamated analog MX-68 (N-[[4-[(2,4-diaminopteridin-6-yl)methyl]-3,4 dihydro-2H-1,4-benzothiazin-7-yl]-carbonyl]-L-homoglutamic acid). METHODS: We studied the effects of low-dose methotrexate (0.75 mg/kg intraperitoneally [IP] every week for 5 weeks), MX-68 (2 mg/kg IP 2 days and 1 hour before induction of inflammation), dexamethasone (1.5 mg/kg IP 1 hour before induction of inflammation), or vehicle control on acute inflammation in an air-pouch model in A(2A) and A(3) receptor knockout mice. RESULTS: Low-dose weekly methotrexate treatment increased the adenosine concentration in the exudates of all mice studied and reduced leukocyte and tumor necrosis factor alpha accumulation in the exudates of wild-type mice, but not in those of A(2A) or A(3) receptor knockout mice. Dexamethasone, an agent that suppresses inflammation by a different mechanism, was equally effective at suppressing leukocyte accumulation in A(2A) knockout, A(3) knockout, and wild-type mice, indicating that the lack of response was specific for methotrexate and MX-68. CONCLUSION: These findings confirm that adenosine, acting at A(2A) and A(3) receptors, is a potent regulator of inflammation. Moreover, these results provide strong evidence that adenosine, acting at either or both of these receptors, mediates the antiinflammatory effects of methotrexate and its analog MX-68. PMID- 12528126 TI - Serum amyloid P component binds to late apoptotic cells and mediates their uptake by monocyte-derived macrophages. AB - OBJECTIVE: Some pentraxins, such as C-reactive protein, bind to apoptotic cells and are involved in the clearance of these cells. We undertook this study to determine whether serum amyloid P component (SAP; a pentraxin that, when deficient in mice, results in lupus-like disease) binds to apoptotic cells and to assess the functional consequences of SAP binding for their phagocytosis by macrophages. METHODS: Human peripheral blood monocytes were isolated and cultured for 7 days to obtain monocyte-derived macrophages. Jurkat cells were irradiated with ultraviolet B to induce apoptosis. After 4 hours, a mean +/- SEM of 54.0 +/- 5.1% of these cells stained with annexin V and were propidium iodide negative (early apoptotic [EA] cells). After 24 hours, 77.3 +/- 2.7% of cells stained positive with both annexin V and propidium iodide (late apoptotic [LA] cells or secondary necrotic cells). EA and LA cells were incubated with fluorescein isothiocyanate-labeled SAP in the presence or absence of Ca(2+), and binding was measured by flow cytometry. Phagocytosis was tested by incubation of macrophages with EA or LA cells in the presence of normal human serum (NHS) and quantified as a phagocytosis index (PI; number of Jurkat cells internalized by 100 macrophages). Experiments were repeated with SAP-depleted serum and after reconstitution with increasing concentrations of SAP. RESULTS: The majority of LA cells did bind SAP in the presence of Ca(2+), whereas EA cells did not. SAP depletion of NHS resulted in a 50% decrease in the PI for LA cells, and complete restoration of the PI could be demonstrated with SAP reconstitution up to 100 microg/ml. SAP depletion had no effect on phagocytosis of EA cells. CONCLUSION: SAP binds to LA cells and is involved in the phagocytosis of these cells by human monocyte-derived macrophages. This may have consequences for diseases such as systemic lupus erythematosus, in which phagocytosis of apoptotic cells is decreased. PMID- 12528127 TI - Coordinate expression of activating Fc gamma receptors I and III and inhibiting Fc gamma receptor type II in the determination of joint inflammation and cartilage destruction during immune complex-mediated arthritis. AB - OBJECTIVE: To study the role of the activating Fc gamma receptor types I and III (Fc gamma RI and Fc gamma RIII, respectively) and the inhibiting Fc gamma receptor II (Fc gamma RII) in inflammation and in various aspects of cartilage destruction during arthritis that is solely induced by immune complexes. METHODS: Immune complex-mediated arthritis (ICA) was passively induced by lysozyme antilysozyme complexes in Fc gamma RI-, Fc gamma RIII-, and Fc gamma RII-knockout mice and their wild-type controls. Total knee joints were isolated to study inflammation and cartilage destruction (loss of proteoglycans [PGs], chondrocyte death, matrix metalloproteinase [MMP]-mediated neoepitope [VDIPEN] expression, and erosion). The presence of an active phenotype of macrophages was studied by detection of myeloid-related proteins 8 and 14 (MRP8 and MRP14, respectively). RESULTS: Influx and activation of inflammatory cells (MRP expression) during ICA was decreased in Fc gamma RIII-deficient mice and enhanced in mice lacking Fc gamma RII. Mild cartilage destruction reflected by loss of PGs was consistent with the degree of inflammation. Mice lacking Fc gamma RIII showed almost no PG depletion, whereas in Fc gamma RII(-/-) mice, PG depletion was increased 3-7-fold in various cartilage areas. Initiation of erosive cartilage destruction, as reflected by MMP-mediated VDIPEN expression, was reduced in Fc gamma RIII(-/-) and Fc gamma RI(-/-) mice, directing the two different critical steps of cellular influx and subsequent activation. These aspects were enhanced in Fc gamma RII(-/ ) mice. In Fc gamma RI(-/-) and Fc gamma RIII(-/-) mice, VDIPEN expression was 90 99% lower, whereas in Fc gamma RII(-/-) mice, VDIPEN expression was increased 4 fold. Chondrocyte death was reduced in Fc gamma RIII(-/-) mice (68% lower) and enhanced in Fc gamma RII(-/-) mice (6-12-fold higher). Progression of arthritis and erosion of the cartilage surface were markedly elevated in Fc gamma RII(-/-) arthritic joints. CONCLUSION: During ICA, Fc gamma RIII is the dominant activating receptor mediating joint inflammation, whereas both Fc gamma RI and Fc gamma RIII are involved in cartilage destruction. Fc gamma RII inhibits both joint inflammation and severe cartilage destruction during ICA. PMID- 12528128 TI - Concomitant disappearance of electrocardiographic abnormalities and of acquired maternal autoantibodies during the first year of life in infants who had QT interval prolongation and anti-SSA/Ro positivity without congenital heart block at birth. PMID- 12528129 TI - Clinical images: Hepatitis C-associated osteosclerosis. PMID- 12528130 TI - Paraplegia in a patient receiving anti-tumor necrosis factor therapy for rheumatoid arthritis: comment on the article by Mohan et al. PMID- 12528131 TI - Unproven hypothesis that leflunomide is better than methotrexate as measured by magnetic resonance imaging: comment on the article by Reece et al. PMID- 12528133 TI - Modified disease activity score versus radiologic progression as an indicator of rheumatoid arthritis disease status: comment on the article by Landewe et al. PMID- 12528135 TI - TNFR2 position 196 polymorphism in Japanese patients with rheumatoid arthritis: comment on the article by Dieude et al. PMID- 12528136 TI - Study of stromal cell-derived factor 1 alpha in synovial fluid in early rheumatoid arthritis: comment on the article by Kanbe et al. PMID- 12528138 TI - Fibromyalgia is not a diagnosis: comment on the editorial by Crofford and Clauw. PMID- 12528139 TI - Pyridostigmine for fibromyalgia: comment on the article by Paiva and a historical vignette. PMID- 12528141 TI - Production of the chemokine RANTES by articular chondrocytes and its role in cartilage degradation: comment on the article by Alaaeddine et al. PMID- 12528142 TI - Willow bark extract, a useful alternative for the treatment of osteoarthritis: comment on the editorial by Marcus and Suarez-Almazor. PMID- 12528144 TI - Prevention of recurrence of congenital heart block with intravenous immunoglobulin and corticosteroid therapy: comment on the editorial by Buyon et al. PMID- 12528146 TI - Use of sildenafil citrate in Raynaud's phenomenon: comment on the article by Thompson et al. PMID- 12528147 TI - A further note on testing for a birth order effect in ankylosing spondylitis. PMID- 12528152 TI - Integrated fetal testing in growth restriction: combining multivessel Doppler and biophysical parameters. PMID- 12528153 TI - Predicting pregnancy failure in 'empty' gestational sacs. PMID- 12528154 TI - Staying the course. PMID- 12528155 TI - Nasal bone hypoplasia in trisomy 21 at 15-22 weeks' gestation. AB - OBJECTIVE: To investigate the potential value of ultrasound examination of the fetal profile for present/hypoplastic fetal nasal bone at 15-22 weeks' gestation as a marker for trisomy 21. METHODS: This was an observational ultrasound study in 1046 singleton pregnancies undergoing amniocentesis for fetal karyotyping at 15-22 (median, 17) weeks' gestation. Immediately before amniocentesis the fetal profile was examined to determine if the nasal bone was present or hypoplastic (absent or shorter than 2.5 mm). The incidence of nasal hypoplasia in the trisomy 21 and the chromosomally normal fetuses was determined and the likelihood ratio for trisomy 21 for nasal hypoplasia was calculated. RESULTS: All fetuses were successfully examined for the presence of the nasal bone. The nasal bone was hypoplastic in 21/34 (61.8%) fetuses with trisomy 21, in 12/982 (1.2%) chromosomally normal fetuses and in 1/30 (3.3%) fetuses with other chromosomal defects. In 3/21 (14.3%) trisomy 21 fetuses with nasal hypoplasia there were no other abnormal ultrasound findings. In the chromosomally normal group hypoplastic nasal bone was found in 0.5% of Caucasians and in 8.8% of Afro-Caribbeans. The likelihood ratio for trisomy 21 for hypoplastic nasal bone was 50.5 (95% CI 27.1 92.7) and for present nasal bone it was 0.38 (95% CI 0.24-0.56). CONCLUSION: Nasal bone hypoplasia at the 15-22-week scan is associated with a high risk for trisomy 21 and it is a highly sensitive and specific marker for this chromosomal abnormality. PMID- 12528156 TI - Factors influencing the prenatal detection of structural congenital heart diseases. AB - OBJECTIVE: To assess the factors influencing the prenatal detection rate of structural congenital heart diseases (CHDs). METHODS: A retrospective study was conducted at a major obstetric hospital in Australia between 1 January 1996 and 30 June 1999. The medical records of all fetuses and infants born with CHD, except those with isolated patent ductus arteriosus or secundum atrial septal defect, were reviewed. Only pregnancies that had prenatal ultrasound scan assessments for morphological surveys were included. The following factors that may influence the detection rate were assessed: complexity of the lesions; experience of the sonographers (performance in tertiary versus non-tertiary institutions); presence of other structural or chromosomal anomalies; and maternal body mass index (BMI). RESULTS: The incidence of structural CHD in this series, excluding cases referred from other hospitals, was 7.0 per 1000 (179/25 529). Of the 179 pregnancies with CHD, 151 had prenatal ultrasound scans and were included in the study. The overall detection rate for CHDs in this series was 40.4%. The detection rate for isolated septal defects was poor (13.7%). The detection rates were significantly higher for complex lesions (54%), for lesions with concomitant septal defects (66.7%), and for lesions with abnormal four chamber views (62.9%). The detection rate was also higher if the scan was performed in the tertiary institution, and if there were other chromosomal or structural anomalies. Maternal BMI did not affect the detection rate in the current series. Stepwise logistic regression analysis showed that three independent variables affecting the detection rate were complexity of the cardiac lesion, experience of the operator, and the detection of chromosomal anomalies. CONCLUSION: A high detection rate for major CHDs can be achieved in a screening setting but there is still room for improvement in scanning skills in the four chamber view and great-artery analysis in both tertiary and local centers. PMID- 12528157 TI - Fetal tissue Doppler echocardiography: detection rates of cardiac structures and quantitative assessment of the fetal heart. AB - OBJECTIVES: Tissue Doppler echocardiography (TDE) has been developed in adult cardiology, but only recently has it been applied to fetal heart studies. We implemented TDE on a high-resolution ultrasound system used for prenatal scanning by changing the Doppler settings, but without specific TDE equipment, to study cardiac tissue motion of normal fetuses in the second and third trimesters. METHODS: Using color Doppler settings optimized for low velocity motion assessment, detection rates of cardiac structures were measured in 47 fetuses. With pulsed Doppler, axial diastolic and systolic cardiac wall velocities were recorded in another 30 fetuses. RESULTS: In all fetuses TDE could detect cardiac wall motion. The right ventricular wall and the level of the atrioventricular valves were most easily seen. Axial contraction velocities of the fetal heart increased with gestational age. CONCLUSIONS: TDE can be used to study normal fetal cardiac function throughout gestation. A regular ultrasound system without additional hardware or software is suitable for screening studies using TDE. PMID- 12528158 TI - Fetal cardiac function assessed by Doppler myocardial performance index (Tei Index). AB - OBJECTIVES: The Tei Index (TI) is a useful, non-invasive, Doppler-derived myocardial performance tool which can be used to assess aspects of systolic and diastolic function. The aim of this study was to determine normal values of fetal left ventricular (LV) TI in second- and third- trimester fetuses and to compare these to other values reported in the literature. METHODS: Doppler waveforms of the LV outflow tracts were obtained in 74 second- and early third-trimester fetuses. The LV isovolumic contraction time (ICT), isovolumic relaxation time (IRT) and ejection time (ET) were measured and the TI calculated using the formula (ICT + IRT)/ET. The literature on LV myocardial function in the fetus was also reviewed. RESULTS: The normal TI in second- and early third-trimester fetuses (18-31 weeks' gestation) was 0.53 +/- 0.13. The ICT was 43 +/- 14 ms, the ET was 173 +/- 16 ms and the IRT was 48 +/- 13 ms. CONCLUSION: The TI can be easily obtained in the fetus without the need for precise anatomic imaging. The TI may be a useful tool to explore fetal myocardial function in different clinical situations. PMID- 12528159 TI - Echocardiographic evaluation of fetal hydrothorax: the effusion ratio as a diagnostic tool. AB - OBJECTIVE: Fetal hydrothorax may lead to hydrops and is associated with mortality as high as 50%. The objective of this study was to define the pathophysiology of fetal hydrothorax and its relation to hydrops. METHODS: Measurements from echocardiograms of 33 fetuses diagnosed with hydrothorax were made, and included diameters of the thorax, heart, inferior vena cava, right ventricle (RV), left ventricle (LV) and aortic and pulmonary valves. Doppler-derived velocities were measured in the aorta and pulmonary artery just above the aortic and pulmonary valves. The ratio of the area of the effusion to the area of the thorax (effusion ratio) was calculated. Variables were converted into Z-scores from regression equations based on normal data. Features of fetuses with and without hydrops were compared. RESULTS: Higher effusion ratios were noted in hydropic versus non hydropic fetuses. Compared to a normal population, study subjects had smaller dimensions of LV, RV and aortic and pulmonary valves. They also had higher pulmonary artery peak velocities. The comparison between non-hydropic and hydropic fetuses revealed lower values for LV and pulmonary valve dimensions, and peak aortic velocity in hydropic fetuses. The severity of LV compression correlated significantly with effusion ratio. CONCLUSION: Fetal hydrothorax is accompanied by compression of the cardiac structures, resulting in altered cardiac hemodynamics. Echocardiographic assessment, including the measurement of effusion ratio, may be a useful tool in guiding fetal therapy. PMID- 12528160 TI - Outcome of children with prenatally diagnosed central nervous system malformations. AB - OBJECTIVE: To study the outcome of pregnancies with a prenatally diagnosed central nervous system (CNS) malformation. METHODS: Leiden University Medical Centre is a tertiary referral center for fetal ultrasound and invasive prenatal diagnosis. Maternal and neonatal records of prenatally diagnosed CNS malformations were retrospectively reviewed over a 6-year period (1993-1998). Information on current development of surviving children was obtained by contacting the care-giving pediatric neurologist. RESULTS: During the study period 124 fetuses were diagnosed with a CNS malformation. Data on pregnancy and delivery were available for 118 pregnancies. Additional malformations were present in 47% of fetuses (55/118). A total of 46% of pregnancies (54/118) were terminated, and 15% (18/118) ended in spontaneous intrauterine death. A total of 39% of pregnancies (46/118) resulted in live birth, and 29 of the infants were still alive at the age of 3 months. One child was lost to follow-up, one infant died at the age of 4 months, and two children died at the age of 3 years. Psychomotor development of the remaining 25 children was normal for five, slightly disabled for seven, moderately disabled for five and severely disabled for eight. CONCLUSION: Due to the high rate of termination of pregnancy and to the frequent association with other anomalies, the survival rate of pregnancies in which a CNS defect had been diagnosed prenatally was only 25%. More than 50% of surviving children were moderately or severely disabled. PMID- 12528161 TI - Audit of prenatal and postnatal diagnosis of isolated open spina bifida in three university hospitals in The Netherlands. AB - OBJECTIVE: To audit the current Dutch policy of prenatal detection of isolated open spina bifida based on offering detailed ultrasound examination only on indication. METHODS: A retrospective analysis of prenatally diagnosed isolated spina bifida cases and of newborns diagnosed with this condition was carried out in three university hospitals. The data were collected from databases and clinical records of the departments of prenatal diagnosis, obstetrics, neonatology, child neurology and neurosurgery of the three centers. RESULTS: Between January 1996 and December 1999, 88 cases of isolated open spina bifida were diagnosed prenatally by ultrasound investigation. Thirty-eight cases (43%) were diagnosed before the 24th week of gestation. Of these, 35 (92%) ended in termination of the pregnancy at the parents' request. Of the remaining 50 cases (57%) diagnosed after the 24th week of gestation, eight (16%) pregnancies were terminated beyond the legal limit for termination due to the severity of the condition. Of the 88 cases of isolated spina bifida, 25 infants (28%) were still alive at the age of 4 years. In the same audit period 112 newborn infants with isolated open spina bifida were admitted to the neonatology, child neurology, or neurosurgery ward of the three centers. Of these cases, 47 (42%) had been diagnosed prenatally and 65 (58%) were an unexpected finding at birth. In 24 infants (21%) surgical treatment was withheld because of the severity of the condition and predicted poor outcome, whereas the remaining 88 infants (79%) underwent surgical repair. CONCLUSION: The current practice in The Netherlands of offering ultrasound screening to high-risk patients only leads to the early detection of a minority of cases of spina bifida. Most cases are diagnosed either after the 24th week of gestation or they remain undiagnosed until after birth. When spina bifida is diagnosed before the 24th week of gestation the vast majority of parents opt for termination. In order to reduce the birth prevalence of spina bifida in The Netherlands the introduction of a policy of routine ultrasound screening should be considered. PMID- 12528162 TI - Changes in flow velocity patterns of the superior and inferior venae cavae during placental circulatory insufficiency. AB - OBJECTIVES: Placental circulatory insufficiency, expressed by the disappearance of the diastolic component of the umbilical artery Doppler velocity waveforms, causes blood flow redistribution that could disturb, to different extents, the systemic venous returns to the heart. The purpose of this study was to investigate the effect of an absence of diastolic blood flow in the umbilical artery on the relationship between the Doppler flow velocities of the venae cavae. METHODS: Fifteen normal fetuses (normal group) were matched for gestational age with 11 fetuses with absent diastolic flow in the umbilical artery (abnormal group). In the venae cavae, the following Doppler variables were measured and compared between groups: (a) during ventricular systole: maximum (S(max)) and minimum velocities (S(min)) and velocity integrals (SI); (b) during ventricular diastole: peak velocity of the E-wave and its integral (EI), the A wave and its integral (AI). A venous velocity index (VVI) was defined as (S(max) + S(min))/S(max). RESULTS: In the normal group, S(min) and VVI were significantly higher in the inferior vena cava (IVC) than in the superior vena cava (SVC). The ratio SVC-VVI/IVC-VVI was therefore always less than one. In the abnormal group, S(min), SI, E, EI and VVI were higher in the SVC compared to those of the IVC. The ratio SVC-VVI/IVC-VVI was always greater than one. CONCLUSION: In the absence of umbilical artery diastolic flow, a reciprocal shift is observed between the IVC and SVC velocity waveforms characterized by a flow profile in the IVC which resembles that of a normal SVC profile and vice versa. These changes are another manifestation of blood flow redistribution towards the brain in the presence of placental circulatory insufficiency. They should be taken into account on Doppler assessment of ventricular diastolic function based on venous flow patterns during placental circulatory impairment. PMID- 12528163 TI - Prediction of early pregnancy viability in the absence of an ultrasonically detectable embryo. AB - OBJECTIVE: To identify clinical, ultrasound and biochemical parameters that may allow prediction of pregnancy viability in women without a detectable embryo on ultrasound examination. METHODS: This was a prospective observational study of pregnant women with an ultrasound finding of a gestational sac measuring < 20 mm mean diameter without a visible embryo. Women's age, menstrual dates, clinical symptoms (pain and bleeding), mean gestational sac diameter and measurements of serum beta-human chorionic gonadotropin and progesterone were recorded in all cases. All women were managed expectantly until the pregnancy viability was established conclusively based on clinical and ultrasound findings. All parameters were tested by univariate analysis and then analyzed in a stepwise procedure to form a logistic regression model for predicting pregnancy viability. RESULTS: One hundred and eighteen (59%) women had a normal intrauterine pregnancy and 82 (41%) had a miscarriage. Stepwise analysis showed that three diagnostic parameters (maternal age, gestational sac diameter and serum progesterone) contributed significantly to the predictive power of the logistic model. With this model, at a cut-off value of 10% probability, the diagnosis of viable pregnancy was made with a sensitivity of 99.2% (95% CI, 95.8-99.97) and specificity of 70.7% (95% CI, 61.3-78.9). CONCLUSION: The use of a logistic regression model allows prediction of pregnancy viability when an embryo cannot be visualized on ultrasound scan. PMID- 12528164 TI - Sonography of pregnancies with first-trimester bleeding and a small intrauterine gestational sac without a demonstrable embryo. AB - OBJECTIVE: This was a prospective observational cohort study to evaluate the outcome and prognostic criteria of pregnancies with first-trimester bleeding and a gestational sac or = 100 ng/mL) and 100 normoprolactinemic patients (PRL 0-25 ng/mL), delta max. PRL (the difference between maximal prolactin (PRL) after thyrotropin releasing hormone (TRH) injection and basal value) was compared with basal PRL and computed tomography (CT) of the sellar region. In 122 hyperprolactinemic patients delta max. PRL was < 100%, while tumor was found in 106 of them. In the remainder seven hyperprolactinemic patients delta max. PRL was > or = 100% and CT showed no tumor. A significant difference in delta max. PRL between hyperprolactinemic patients without and those with verified adenoma was found and showed a significant negative correlation with basal PRL. Between 122 hyperprolactinemic patients with delta max. PRL < 100%, mean basal PRL and duration of clinical symptoms were significantly lower in 16 patients with normal CT compared to 106 patients with tumor. All normoprolactinemic patients showed delta max. PRL > or = 100% and no tumor on CT. PRL stimulation disturbance precedes tumor visualization and represents a decisive diagnostic parameter in hyperprolactinemic patients with no tumor signs. PMID- 12528294 TI - Lymphocytic adenohypophysitis simulating a pituitary adenoma in a pregnant woman. AB - The lymphocytic hypophysitis, appearing in women during the third trimester of pregnancy or early post-partum period, is a rare cause of hypopituitarism and pituitary enlargement. A 39 year-old woman presented in the 37th week of pregnancy with bilateral heteronymous quadrantanopsia, CT indicative of tumorous mass and symptoms of hypopituitarism with decreased thyroid hormone and thyrotrophin levels, and low normal level of cortisol. After the birth of a healthy male child the patient breastfed for 10 days, sight disturbances disappeared, but amenorrhea persisted. Upon admittance the visual field showed no abnormalities. MR of the sellar region confirmed previous CT findings. Endocrinological testing confirmed secondary hypothyroidism and cortisol deficiency, normal levels of prolactin with satisfactory reaction to thyroliberin. Histology showed mononuclear infiltration, and immunohistochemistry revealed T-cells (CD3) at the borders, and B-cells (CD20) in the follicular center. Due to enlargement of the pituitary associated with hypopituitarism, an incorrect diagnosis of a tumor could be made. PMID- 12528295 TI - Tooth morphology in function of selfprotective mechanism. AB - The degree of curvature of vestibular and oral tooth surfaces is determined by the shape sequence from the tooth crown to the epithelial attachment and the alveolar bone, and directly influences the health of gingiva and the entire tooth support system. The goal of this research was to determine vestibulo-oral planes of upper and lower permanent premolars and molars and the thickness of the associated alveolar osseous wall, and 2,727 measurings were processed using pertinent statistical procedures. The results have shown the degree of curvature of vestibular and oral tooth surfaces to be higher in upper teeth than in lower ones (p > 0.05); vestibular convexities higher than oral ones and especially noticeable at the junction from the meandle to the lower third of the crown. Higher vestibular curvature also entailed thicker osseous wall. All results were higher than the ones found in literature. We consider our results to be relevant for our population. The research on the relationship of teeth and the alveolar bone should be continued by using even more test points and more sophisticated research procedures. PMID- 12528296 TI - A method to evaluate and compare two different intraoral radiographs of the same patient. AB - Objective of this study was to determine the accuracy of the method of the clinical intraoral densitometry, to compare differences in the calculation with or without subtraction of the background adjacent soft-tissues from the stepwedge (SW) and to verify which regression model best fitted the experimental data in order to express the measured values in equivalents of SW thickness. Two intraoral radiographs, one after another, were made for each of 6 patients. A copper SW (6 steps, thickness 0.05-0.3 mm) was attached to each radiograph, trying to avoid the superimposition of the bony structures. Films were processed and digitized. Grey levels were measured on each step of the SW, on the background of the SW and on the same 3 randomly chosen regions of interest (ROIs) on each digitized image. The measurement with and without the subtraction of optical densities of the background around the SW from the optical densities of the SW was performed. For the calculation of the SW thickness equivalents, the regression analysis was performed by using different regression models. The best fitting regression model was the 3rd degree polynomial. The results were more precise when using the subtraction of the background overlapping the SW. PMID- 12528297 TI - Double tooth. AB - The form of primary and permanent teeth can differ morphologically from that which is considered normal, completely or in some parts. The changes in tooth form can be hereditary or caused by some disease or trauma. Fusion is a union of one or more teeth during development. Gemination means that two separate morphological units were created by division of the tooth germ. The intention of this study was to state the prevalence of double teeth (fusion and gemination) among the persons tested, as to gender, distribution in the maxilla or mandible, and whether the anomaly occurred bilaterally or unilaterally. The results of this investigation have shown that in a total of examined 3,517 plaster models, a prevalence of double teeth was 0.2%. 57.2% of them were fusioned and 42.9% geminated. PMID- 12528298 TI - Screening of fixed prosthodontic dentures after five years of use in relation to material and construction. AB - The aim of this study was to screen patients with fixed prosthodontic appliances that were in oral cavity for a period of 5 years or more and to assess clinically and radiologically root caries, gingival recession, pocket formation, alveolar ridge resorption, as well as gingival (GI) and plaque index (PI). The aim also was to find out the differences between materials and constructions, between abutment and non-abutment teeth, and to find out the need for replacement. A total of 260 patients and their orthopantomograms were examined, with a total of 2,265 teeth, 610 being bridge abutments and 246 being crowns. The most frequent were metal+ acrylic veneer crowns or bridges. Root caries was found under the abutments in 10-20%; however abutments with ceramic crowns had the lowest percentage of caries (p < 0.01). Alveolar ridge resorption, pocket formation deeper than 3 mm and gingival recession of various degree was found in 50% of the cases, again with the lowest percentage of ceramic-fused-to-metal appliances (p < 0.01). Pocket depth was registered in significantly higher percentage in metal acrylic veneer appliances compared to natural teeth (p < 0.01), while there was no significant difference between metal-ceramic appliances and natural teeth (p > 0.05). Although the worst findings were recorded for metal- + acrylic veneer crowns for PI, no significant difference existed between crowns of different material or non-abutment teeth (p > 0.05). There was statistically significant difference between abutments with metal + acrylic veneer crowns, full metal crowns, metal ceramic crowns and non-abutments for GI scores. Higher percentage of scores 0 and 1 was recorded for metal ceramic crowns and non-abutments and significantly higher percentage of scores 2 and 3 was recorded for metal + acrylic veneer crowns and full metallic crowns. Almost 50% of metal-ceramic abutments had no pathologic findings. Almost 30% of the patients needed replacement, or even some abutments to be extracted and therefore a new prosthodontic appliance. PMID- 12528299 TI - Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry detecting NOS in healthy and chronically inflamed pulp. AB - The aim of this study was to examine the expression of nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) activity in human dental pulps and determine whether there are changes of the activity in chronically inflamed pulp tissue. Nineteen pulps with clinical diagnosis of chronic pulpitis were collected during endodontic treatment. The healthy controls were obtained from teeth extracted for orthodontic therapy. The clinical diagnosis was confirmed by histological analysis. Healthy pulps showed stratified odontoblasts in peripheral parts, while in central area there was normal connective tissue. Chronically inflamed pulps showed less expressed stratification of odontoblasts and infiltration of lymphocytes, polymorphonuclear leukocytes, plasma cells and mastocytes. NADPH-d granular reactivity was assessed semi quantitatively under the light microscope by a single observer and scored on an intensity scale from negative reaction to very strong reaction. In healthy human pulps, NADPH-d activity was strong to very strong in odontoblastic layer. Endothelial cells and Schwann cells showed strong NADPH-d reactivity, while the other parts of central area were weakly positive. Similar distribution of reactivity was expressed also in chronically inflamed pulp; moderate to strong reaction was observed in stromal area as result of positive reaction in inflammatory cells and endothelial cells of abundant newly formed capillaries. PMID- 12528300 TI - Oral status, aesthetic materials and frequency of crowns and bridges in patients with fixed prosthetic appliances living in the Metkovic region. AB - The aim of the study was to examine oral status in patients of the Metkovic region with fixed prosthodontic appliances. The aim was also to evaluate aesthetic materials used, and difference between the frequency of crowns and bridges dependent on patient's age and frequency of visits to dentist. The examination was performed on 212 dental patients with fixed prosthetic appliances, from the Public Health Center "Metkovic". A questionnaire was created for the purpose of this study and was filled in by the patients. The dentist registered oral status. Descriptive statistics and chi 2 tests were used for the data analysis. Following conclusions were made: 1. The examined patients had relatively high caries (tooth decay), gangrene and residual roots prevalence, the oral hygiene was not appropriate, calculus and gingivitis were diagnosed in all patients (100%) and these facts point out to the insufficient preventive oral health care in the Metkovic region, as well as to the insufficient self perception on the proper oral hygiene maintenance. 2. Patients who visit their dentist regularly (once a year or more) have significantly more crowns than bridges than the patients who visit their dentist irregularly or when in pain (p < 0.01). 3. Almost all fixed prosthetic appliances not older than 10 years were made of porcelain (98%), while acrylic veneer crowns (or chromasite) were more frequent in appliances older than 10 or 15 years (p < 0.01). There was no gender difference dependent on the material used (p > 0.05). High frequency of porcelain was attributed to the high economic status of this region, as well as to a dentist's preference. PMID- 12528301 TI - The ethical structure of Homeric society. AB - According to Plato, the society as reflected in Homer's epics is one without state power. In a society without state power, the act of revenge which the offended party prosecutes on the offender is the only disciplinary force, which should be endorsed by the ethical concepts of the society. Though the ethics of Homeric society has been analyzed before, there has been no theory on the ethical structure of Homeric society analyzed from the viewpoint of a society without state power. This study attempts to address this issue. Six concepts, "oath", "honor", "guest", "blood", "food" and "revenge" have been extracted from Homeric epics in comparison with the ethical structure of the Albanian Kanun. The ethical structure of Homeric society appears to belong to the category of the ethics of a society without state power, as represented by the ethical structure of the Kanun. Plato explicitly criticizes the ethical value system of Homeric society, which he thinks is alienated from the idea of Good. PMID- 12528302 TI - Antibacterial agents: patent highlights January to June 2002. AB - A total of 48 patents dealing with disclosures on the different classes of antibacterial agents, including the beta-lactams, oxazolidinones, macrolides, quinolones, tetracyclines and miscellaneous antibacterial agents reported between January and June 2002 are selected for review. The miscellaneous agents section focused on the significant discovery of potential lead compounds as inhibitors of bacterial fatty acid synthase and peptide deformylase, and also included examples of novel peptidic antibiotics and pleuromutilin derivatives along with their antibacterial activities. Only a few patents disclosed novel agents in the quinolone and carbapenem areas. There are several patents disclosing improved formulation of old agents to increase their effectiveness and stability upon storage. Two patents disclosed effectiveness of antibiotic combinations with respect to the newer antibiotics linezolid and quinupristin/dalfopristin. PMID- 12528303 TI - Novel targets for antibiotic drug design. AB - The urgent need for new antibiotics has led to an explosion in the number and diversity of antibiotic drug targets under investigation. The majority of such targets are enzymes required for essential cellular functions. Often, such novel targets are completely unexploited for antibiotic therapy and therefore have the advantage of avoiding current resistance mechanisms. In general, the most advanced novel targets are drawn from processes where an existing antibacterial compound has validated that process for antibiotic therapy. This review describes a number of the more promising targets under development. PMID- 12528304 TI - Heat shock proteins in stroke and neurodegenerative diseases. PMID- 12528305 TI - Neuro-inflammation as a therapeutic target in amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a rapidly progressive neuromuscular disease that destroys both upper and lower motor neurons, resulting in spasticity, diffuse muscular atrophy, weakness, and ultimately death from respiratory failure. It is presumed that in the vast majority of cases, ALS is acquired and occurs sporadically, although the exact etiology is unknown. Recent, emerging evidence suggests that neuro-inflammation may be a pathological characteristic of this disease; this could represent a potential therapeutic target for a pharmacological agent to help treat this severe disease. This article reviews the current research in this area and discusses theoretical and clinical ramifications of these recent findings. PMID- 12528306 TI - Anti-inflammatory strategies in coronary atherosclerosis. AB - Recent evidence has demonstrated that inflammation plays an important role in the progression of coronary atherosclerosis. This review focuses on acute coronary syndromes and examines some novel therapeutic strategies aimed at manipulating the inflammatory environment in these patients in order to reduce the subsequent major adverse coronary event rate following myocardial infarction. PMID- 12528307 TI - Third generation thrombolytics for the treatment of ischemic stroke. AB - The field of intravenous and intra-arterial thrombolysis for the treatment of acute ischemic stroke is rapidly advancing. Limitations of existing thrombolytic agents have prompted the development of new thrombolytic agents over the last decade. These new agents are broadly classified as third generation thrombolytics. Two of the several third generation thrombolytic agents have been investigated for the treatment of acute ischemic stroke and include tenecteplase (Genentech Inc) and reteplase (Roche Holding AG). By virtue of structural modifications, third generation thrombolytics have longer half-lives and greater penetration into the thrombus matrix. Tenecteplase has been evaluated in experimental models of ischemic stroke. These experimental studies have observed faster and more complete recanalization of occluded arteries compared with second generation thrombolytics. The first prospective human clinical trial evaluated the safety and efficacy of intra-arterial reteplase in 16 patients with ischemic stroke who were poor candidates for intravenous alteplase therapy. Near complete or complete recanalization was observed after treatment in 88% of the patients. The development and use of third generation thrombolytics is expected to increase the rate of recanalization and clinical recovery in patients with ischemic stroke. Clinical trials are required to determine the appropriate dose and patient selection for these emerging pharmacological agents. PMID- 12528308 TI - BIII-890-CL. Boehringer Ingelheim. AB - BIII-890-CL is a non-orally active sodium channel blocker under development by Boehringer Ingelheim Corp for the potential treatment of thromboembolic stroke. By April 2001, the compound was in phase II clinical trials for this indication, with trials ongoing in June 2002. The compound is also under investigation for its potential use in the treatment of pain. PMID- 12528309 TI - ZK-807834. Berlex. AB - Berlex (a Schering subsidiary) and Pfizer (formerly Parke-Davis) are developing ZK-807834 (CI-1031), one of a series of Factor Xa inhibitors, for the potential parenteral treatment of unstable angina pectoris. PMID- 12528310 TI - Translation of developmental biology into therapeutics: the hedgehog signaling pathway and neurodegenerative diseases. PMID- 12528311 TI - Caspases and neuroprotection. AB - Apoptotic cell death has been implicated in the pathogenesis of both acute and chronic neurodegenerative disorders. The caspase family of cysteine proteases are involved both in the initiation and final execution of apoptosis. Inhibition of the caspase family prevents cell death in a number of models of neurodegenerative cell death in vivo and in vitro. This sparing of neurons does not always correlate with long-term functional recovery, possibly due to the limitations of the available inhibitors. In this review, the evidence for a neuroprotective role of caspase inhibition in models of Parkinson's disease and cerebral ischemia is critically evaluated. PMID- 12528312 TI - Neurotrophins, neuroprotection and the blood-brain barrier. AB - Small molecule drugs have not been effective neuroprotective agents in either the acute treatment of stroke or the chronic treatment of neurodegeneration. Thus, it is time to consider large molecule drugs such as recombinant neurotrophins. However, like many of the small molecules, neurotrophins do not cross the brain capillary endothelial wall, which forms the blood-brain barrier (BBB). Neurotrophins can be made transportable across the BBB by using chimeric peptide brain drug targeting technology, in which a neurotrophin is reformulated by fusion to a transport vector. The latter is a peptide or peptidomimetic monoclonal antibody that undergoes receptor-mediated transcytosis through the BBB, and acts as a 'molecular Trojan horse'. Neurotrophin chimeric peptides are highly neuroprotective following delayed intravenous administration, in both global and focal brain ischemia. PMID- 12528313 TI - NXY-059. Centaur. AB - Centaur and AstraZeneca are developing NXY-059, an alpha-phenyl-N-tert-butyl nitrone derivative, free radical trapping agent, for the potential therapy of reperfusion injury following stroke. Phase II clinical trials are ongoing, with phase III trials in acute stroke planned for initiation in early 2003. PMID- 12528314 TI - CPI-1189. Centaur. AB - The efficacy of the proapoptotic cytokine tumor necrosis factor (TNF) alpha inhibiting compound CPI-1189 has been demonstrated in various cell culture and animal models of chronic neurodegenerative and inflammatory diseases. CPI-1189 intracellularly inhibits the p38 mitogen-activated protein kinase phosphoactivation, thereby protecting against TNF alpha-induced neurodegeneration. Clinical proof-of-concept phase IIa trials in patients with Parkinson's disease and AIDS dementia complex were successful. These studies demonstrated clinical relevance for treatment with CPI-1189 (50 to 100 mg/day), which attenuated the deterioration in cognitive and/or motor function without any relevant side effects. Since the importance of neuroprotection is emerging, in particular in neurodegenerative diseases with concomitant observed immunological pro-apoptotic alterations in the central nervous system, long-term application of CPI-1189 could represent a promising future therapeutic alternative, in addition to neuroprotective compounds such as selegiline. PMID- 12528315 TI - Selectively replicating adenoviruses for cancer therapy: an update on clinical development. AB - Adenoviruses can be engineered to replicate selectively in tumor cells but inefficiently in normal cells. ONYX-015 (CI-1042, dl1520; Onyx Pharmaceuticals Inc), which replicates selectively in cells deficient in the p53 pathway, was the first such adenovirus to reach clinical testing. Multiple trials of ONYX-015 in over 300 cancer patients, and trials with other selectively replicating adenoviruses, have established the safety of this approach. Evidence of anticancer activity in patients is encouraging. Recently, the first clinical trial of a selectively replicating adenovirus carrying an inserted transgene was reported. Adenoviruses with improved efficiency of replication, technologies for use of the viruses as vectors for anticancer gene therapy, and various other approaches, provide promising directions to develop selectively replicating adenoviruses into systemic therapy for metastatic cancer. PMID- 12528316 TI - INGN-241. Introgen. AB - Introgen, under license from Corixa (formerly GenQuest), is developing INGN-241, a gene therapy based on the mda-7 gene in combination with the company's adenoviral delivery system, for the potential treatment of cancer. A phase I trial for the treatment of solid tumors was initiated at the end of November 2000. PMID- 12528318 TI - Side effects of statins--prevalence, relevance, consequences. PMID- 12528317 TI - How to induce severe electrolyte disturbances in the elderly? PMID- 12528319 TI - [National Health Care Plan for Austria: development and critical considerations]. AB - Compared to health services in other countries in the world, the Austrian health care system works relatively well. At the moment it is shaped and dominated by various structures, decision makers and financial backers, which means that it does not work as well as it could under ideal circumstances. The government now wants to develop a national health care plan which, in a concerted action by all those involved in the health service, shall guarantee superior medical care while at the same time taking into account economic aspects. This paper takes a critical look at the idea of an Austrian health care plan from a clinical point of view. PMID- 12528320 TI - Giant platelets in idiopathic thrombocytopenic purpura. PMID- 12528321 TI - Outcome of conservative medical treatment of patients with infrarenal abdominal aortic aneurysms. AB - OBJECTIVE: Since non-invasive diagnostic methods have become available and screening programs have become popular, abdominal aortic aneurysms are more frequently being detected at an early stage of the disease. We analyzed the course of conservatively treated patients with infrarenal abdominal aortic aneurysms (AAA), and determined independent risk factors for aneurysm expansion. METHODS: The study was designed as a retrospective-cohort study including 110 consecutive patients with AAA. Cardiovascular risk factors, comorbidities, current medication, and the findings of color coded duplex sonography and computed tomography were recorded. Ninety-two conservatively treated patients were re-investigated every 6 or 12 months (depending on an initial aneurysm size of > 45 mm or < 45 mm respectively) after initial detection by color coded duplex sonography. We performed a multivariate Cox regression analysis to determine independent predictors of diameter progression (diameter increase > or = 5 mm). RESULTS: We found AAA expansion in 46 conservatively treated patients (50%) during the median follow up period of 23 months (IQR 13 to 33), but no rupture occurred. Baseline diameter > 45 mm (HR 2.3, 95% CI 1.0 to 5.3, P = .04) and signs of aortic dissection in duplex ultrasound (HR 2.2, 1.0 to 4.6, P = .04) were independently associated with aneurysm expansion. The presence of an intraluminal thrombus showed a trend towards higher rates of disease progression (HR 2.6, 95% CI 0.9 to 7.6, P = .08). CONCLUSION: Patients with an aneurysm diameter > 45 mm or ultrasound signs of aortic dissection have an increased risk for AAA progression and need careful evaluation, optimization of risk factors and close (six-month) follow-up intervals. For patients with an aneurysm diameter below 45 mm and without additional risk factors, follow-up intervals of 12 months seem to be safe. PMID- 12528322 TI - [Slow-release morphine hydrochloride for maintenance therapy of opioid dependence]. AB - INTRODUCTION: In Austria, methadone, buprenorphine as well as oral slow-release morphine are used for the treatment of opioid dependence. This controlled examination marks the first time that oral slow-release morphine hydrochloride is applied for maintenance therapy in opioid dependent subjects. METHODS: In order to evaluate the effectiveness of this psychopharmacological medication, we examined patients over a three-week period. Outcome measures were retention rate, additional consumption and the evaluation of opioid withdrawal 24 hours after the last oral medication. RESULTS: Sixty-seven patients were included; sixty-four patients completed the study, representing a retention rate of 94%. During the three-week period, a significant improvement in well-being and a significant reduction in heroin, cocaine and benzodiazepine craving (p < 0.0001) was evaluated. Furthermore, there was a significant reduction of additional consumption of benzodiazepines in supervised urinalysis. Additional consumption of cocaine remained unchanged. Laboratory results showed a significant reduction of CK over the course of investigation. DISCUSSION: The high retention rate of 94% implies a good acceptance and efficacy of the substance. The reduced CK is consistent with a reduction in intravenous application of illegal substances. However, randomized double-blind, double-dummy studies with oral slow-release morphine are needed in order to meet criteria for evidence based medicine. PMID- 12528323 TI - [The frequency of seizures in patients with primary brain tumors or cerebral metastases. An evaluation from the Ludwig Boltzmann Institute of Neuro-Oncology and the Department of Neurology, Kaiser Franz Josef Hospital, Vienna]. AB - Epileptic seizures are common in patients with cerebral metastases as well as in patients with primary brain tumors. In cancer patients without primary brain tumors or brain metastasis, epileptic seizures may occur due to metabolic or toxic causes, or due to infections. We performed a retrospective analysis from our neurooncological database concerning the occurrence of seizures in patients with primary brain tumors, patients with cerebral metastases and in cancer patients without brain tumors. Patients with low grade gliomas, such as astrocytoma WHO I + II (69%), oligodendroglioma WHO II (50%), and mixed glioma WHO II-III (56%) were more likely to have seizures than patients with anaplastic glioma WHO III (44%), glioblastoma WHO IV (48%) or meningeoma (45%). In patients with brain metastasis, melanoma (67%), cancer of the lung (29%), and gastrointestinal tumors (21%) were the primaries with the highest frequency of seizures. In cancer patients without brain metastases or primary brain tumors, seizures occurred in 4%. In conclusion, the occurrence of epileptic seizures in patients suffering from primary brain tumors, as well as in patients with cerebral metastases, varied within the tumor entity. Therefore, especially in brain tumors where a higher probability of epileptic seizures is expected, they should be taken into account in the care of cancer patients. PMID- 12528324 TI - Clinical signs of pulmonary congestion predict outcome in patients with acute chest pain. AB - BACKGROUND: Pulmonary congestion is associated with poor outcome in patients with acute coronary syndromes. In consecutive patients presenting with acute unexplained chest pain to a primary care facility, the prognostic impact of pulmonary congestion is indeterminate. Therefore, we assessed the predictive value of clinical signs of pulmonary congestion in patients presenting with acute chest pain to an emergency department with regard to the origin of the symptoms. METHODS: 1288 consecutive patients with acute chest pain were prospectively assessed for clinical signs of pulmonary congestion. The diagnosis was confirmed by chest radiography. The association of pulmonary congestion and short- and intermediate-term mortality in patients with coronary (n = 381) and non-coronary (n = 907) causes of chest pain was determined using multivariate Cox regression analysis. RESULTS: 108 (8%) patients had clinical signs of pulmonary congestion. Within the mean follow-up period of 23 months (SD 4) 67 patients died, mainly within the first 6 months. Of 108 patients with pulmonary congestion, 82 (76%) had coronary and 26 (24%) had non-coronary chest pain. Pulmonary congestion was independently associated with mortality in patients with coronary chest pain (hazard ratio 6.4, 95% confidence interval 2.5 to 16.1, p < 0.0001), both in patients with acute coronary syndromes or angina pectoris. However, in patients with non-coronary chest pain we observed no independent association of pulmonary congestion with outcome. CONCLUSION: Clinical signs of pulmonary congestion indicate an increased risk for poor outcome in patients with chest pain due to myocardial ischemia. Mortality of these patients is high, particularly in the first months after presentation. Therefore, hospital admission is warranted, including patients with angina pectoris, who otherwise may be candidates for early discharge. PMID- 12528325 TI - Enteral application of an immunoglobulin-enriched colostrum milk preparation for reducing endotoxin translocation and acute phase response in patients undergoing coronary bypass surgery--a randomized placebo-controlled pilot trial. AB - OBJECTIVE: To evaluate the influence of enteral application of an immunoglobulin enriched bovine milk preparation on endotoxin plasma levels, endotoxin neutralizing capacity of plasma (ENC) and the acute phase response (IL-6, CRP) during and after cardiac surgery, in a pilot study. DESIGN, PATIENTS AND METHODS: 60 patients who underwent coronary bypass operations, were evenly enrolled in a placebo-controlled randomized study. The patients were treated by enteral application of either 42 g of a bovine colostrum milk preparation per day or placebo, for 2 days preoperatively. Endotoxin and ENC were sequentially determined intra- and postoperatively by a chromogenic modification of the limulus amebocyte lysate test. Interleukin-6, CRP, transferrin, alpha-2 macroglobulin, albumin, apo-A, apo-B, IgG, IgA, IgM were determined by ELISA and nephelometrically. The clinical course was followed up by daily evaluation of the Apache-II-score. MAIN RESULTS: Demographic data were comparable in both groups. No differences of the Apache-II-score (verum group: 6.5 +/- 1.9 vs. controls: 6.8 +/- 1.8 on admission) were observed. Endotoxin and ENC levels were elevated at the end of the operation and seemed to have a trigger function for the acute phase response. However, there was no reduction (calculated as the area under the curve) in patients receiving the colostrum milk preparation throughout the observation period. Plasma levels of endotoxin binding proteins did not differ. Plasma IL-6 concentrations increased to maximal median values of 655 pg/ml in the verum and 786 pg/ml in the control group, respectively 2 and 6 h after surgery. In the colostrum group, there was a tendency to reduced IL-6 levels throughout the observation period. CRP-levels of all patients peaked 48 h after the operation but were lower (p = 0.034) in the verum group. CONCLUSION: This study revealed that endotoxemia occurs early in an elective non-intestinal surgical intervention, and is followed by a subsequent increase in mediators of the acute phase reaction. The prophylactic enteral application of a bovine milk preparation for two days in cardiac patients did reduce postoperative CRP-plasma levels but, contrary to a former double-blind study in abdominal surgery, failed to curtail perioperative endotoxemia. One reason could be the amount of colostrum preparation administered was too small. PMID- 12528326 TI - Measurement of IgE antibodies using liquid allergens--an inter-method and inter laboratory quality assessment. AB - BACKGROUND: The determination of IgE antibodies is important for the in vitro diagnosis of allergic diseases. However, not all systems currently available in the market fulfill essential quality criteria, e.g. regarding characteristics such as sensitivity and specificity, and the data do not always reflect true clinical relevance in the required fashion. Recent innovations may reduce the workload for the technician, and thus help save time and money. More importantly, they might reduce potential sources of error. OBJECTIVE: Two allergy systems, the well established Pharmacia CAP system that uses the allergens conventionally in a solid phase and the ALLERgen system that employs liquid allergens, were compared with regard to quality criteria and practicability. METHODS: Defined serum pools were checked for within-run and between-days imprecision of IgE antibody detection in two independent laboratories. Serum specimens from allergic patients and controls were tested in parallel using both methods for total and antigen specific IgE antibody detection under standardized conditions. In addition, one laboratory working exclusively with the ALLERgen system participated in the Austrian inter-laboratory quality assessment program. RESULTS: The two systems were comparable in terms of sensitivity and specificity, and also showed good correlation. Within-run evaluations were excellent for total IgE and antigen specific IgE, and the between-days imprecision was satisfactory. Coefficients of variation were within an acceptable range for the different groups of allergens. In the external quality control program the data obtained with the ALLERgen system showed good concordance with other systems in use; up to 94% of the results were identical when considering clinically relevant sensitizations. Regarding practicability, both systems were most satisfactory for the operator. The ALLERgen system offered a certain advantage in terms of automated operation, which resulted in shorter fixed and variable phases of personnel time. CONCLUSION: Both the Pharmacia CAP system and the ALLERgen system belong to an advanced generation of allergy test systems and are easy to handle. The reproducibility of results is good with both methods, and the imprecision data fall within an acceptable range. Thus, the ALLERgen system is a reliable in vitro system for evaluating specific and total IgE in serum, providing data equivalent to those obtained with the CAP system. PMID- 12528327 TI - Severe electrolyte disturbances and renal failure in elderly patients with combined diuretic therapy including xipamid. AB - Diuretics are among the most frequently prescribed substances in elderly patients, but they are also associated with the highest incidence of adverse effects in this group of patients. Xipamide is a sulfonamide-like diuretic whose action does not depend on transtubular secretion. This characteristic makes it suitable for situations in which the kidney is highly sodium avid. Because of the potency of this substance the risk of adverse reactions like electrolyte disorders or hypovolemia is increased as well. We report seven patients (age 65 85) admitted to the emergency room of the University Hospital of Innsbruck between 1998 and 2002 who had developed serious adverse reactions upon initiation of treatment with xipamide as an additional diuretic. Six of these patients had received combinations with loop diuretics. The disturbances observed were hyponatremia (lowest value 108 mmol/l), hypokalemia (lowest value 1.5 mmol/l) and prerenal azotemia (highest serum urea 269 mg/dl, highest serum creatinine 5.13). CONCLUSION: With the exception of diuretic resistance in severe heart failure or renal insufficiency a combination therapy of xipamide with a second diuretic appears to be associated with an unnecessarily high risk of serious adverse reactions and thus should be avoided. This is especially true for elderly patients. PMID- 12528328 TI - Statin-induced myositis migrans. AB - Muscle related side effects of statins might be more frequent than once considered. Without affecting CK, they appear in various forms. Here, for the first time, we describe a new appearance we call "Statin-induced myositis migrans" in a 74-year-old female. Symptoms appeared more during the day and were not related to exercise. They were seen after simvastatin, pravastatin and--most intensively--after atorvastatin, but not on lovastatin, which the patient took for about 7 years without any problem. The clinical consequence (to withdraw the drug or not) of these milder statin side effects are discussed. PMID- 12528329 TI - A case of severe C4d-positive kidney allograft dysfunction in the absence of histomorphologic features of rejection. AB - Acute antibody-mediated (humoral) renal allograft rejection has emerged as a clinicopathological entity that carries a poor prognosis. Its diagnosis is based on typical pathohistologic features, serologic detection of donor-specific alloantibodies and the immunohistochemical finding of endothelial deposits of the complement split product C4d. We herein report a case of severe antibody-mediated graft injury after spousal-donor kidney transplantation. Despite an increased risk for humoral presensitization in the female recipient (three previous pregnancies), donor-specific alloantibodies were not detectable before transplantation. After initial graft function, severe graft dysfunction occurred one week after transplantation. A renal biopsy revealed no histomorphologic features of rejection. However, immunohistochemical detection of diffuse C4d deposits along peritubular capillaries suggested acute humoral rejection. The diagnosis of antibody-mediated rejection was confirmed by the detection of de novo production of anti-donor alloantibodies. Graft dysfunction was resistant to high dose steroids or antilymphocyte antibody therapy. However, a recovery of graft function could be achieved by antibody elimination using immunoadsorption therapy. This case reinforces the high diagnostic value of C4d staining. In severe graft dysfunction humoral immune mechanisms should be considered, even when histopathologic features of humoral rejection are completely absent. PMID- 12528330 TI - [Euthanasia and the paradox in delegation of life assessment]. AB - Active euthanasia, a question of individual existence, fundamentally concerns the concept of human society. Although legalizing voluntary euthanasia seems to be consequent in liberal democracy, delegation of the act of homicide leads to a paradox in liberalism: Generalization in the assessment of individual life--as it is necessary in such a context--undermines the fundamental right of the individual for social existence irrespective of any assessment whatsoever. This is reflected by the humanistic principle of human dignity as it has been described by Kant. Conventional jurisdiction, which permits passive and indirect euthanasia but prosecutes active homicide, avoids this fundamental problem. Nevertheless it facilitates a humane demise within the framework of adequate hospice care. PMID- 12528331 TI - Pulsed magnetic field therapy for osteoarthritis of the knee--a double-blind sham controlled trial. PMID- 12528332 TI - [Does the decree 25/2002 still guarantee the health care of workers exposed to lead?]. PMID- 12528333 TI - [Selection criteria of mobile lifters in the hospital setting]. AB - The manual handling of patients with limited mobility represents the major cause of musculoskeletal injury to the spine in paramedical health care workers. Within the hospital, the more complex procedures of patient transfer often require the use of mobile hoists. The aim of this paper is to describe the basic criteria for the selection of such hoists. The main characteristics of a hoist are its stability, the sling attachment, the speed of operation, range of movement of the spreader bar, safety of the operation being performed, patient comfort, the physical effort required on the part of the health care worker, manoeuvrability and simplicity of use. Important organizational-structural features to evaluate include: the type of patient normally present in the unit concerned, the specific movement to be performed, the structural characteristics of the environment, and the work organization of the personnel. PMID- 12528334 TI - [Cost assessment of one-day stay in the Day Hospital system for motor rehabilitation]. AB - This study develops a model for determining the standard cost of a day's admission to a Day Hospital in a rehabilitation unit. The task is carried out by analysing the costs in a single hospital. The method chosen to determine the standard cost of a day's admission to the Day Hospital was to evaluate the patient's management course. The analysis was limited to the period between the moment the patient is contacted for Day Hospital treatment up to end of the day spent in the Day Hospital. The work has four stages: 1. definition of the patient's management course; 2. identification of all the activities carried out and all the resources involved in the different stages of the management course; 3. evaluation of the resources used by the activities identified, 4. analysis of the clinical records to verify that the management actually provided did not differ from the defined one in order to determine the standard cost. PMID- 12528335 TI - [Promoter effect of mercury chloride and methyl-mercury on human keratinocytes in culture]. AB - Mercury has received considerable media focus because it is present in dental amalgams and seafood. There is potential exposure in gas meters, thermometers and fluorescent lamps workers. To evaluate its possible epigenetic carcinogen effect, cultures of human keratinocytes were treated with increasing doses of HgCl2 for 30 min, 24 h and of CH3HgCl for 24 h, respectively. The red neutral method was used to evaluate the doses of HgCl2 and CH3HgCl which had no cytotoxic effect. Then, the dye transfer method was used to investigate the gap junctions-mediated intercellular communication (GJIC). Cells were microinjected with Lucifer Yellow CH by using the Eppendorf Apparatus and the Leica inverted microscope. After 30 min incubation at the concentration of 10 microM, HgCl2 did not exert inhibition of GJIC. Conversely, after 24 h at the concentration of 10 nM, HgCl2 inhibited GJIC. Incubation with CH3HgCl at the concentration of 250 nM for 24 h reduced the number of fluorescent cells, thus denoting a inhibition of GJIC. Taken together our data demonstrated that: i) HgCl2 and CH3HgCl exerted an inhibitory effect upon GJIC; ii) HgCl2 resulted to inhibit GJIC at concentrations 25 folds lower than CH3HgCl. Further studies will be addressed to evaluate whether the reversal of GJIC inhibition could be obtained by withdrawal of toxic substance, or by the addition of a GJIC activator like the retinoic acid. Finally to shed light on the possible effect of mercury derivates at the transcriptional or translational levels, the expression profile of the connexin 43 gene after HgCl2 and CH3HgCl exposure of cultured human keratinocytes will be investigated. PMID- 12528337 TI - [Rationale for the building of a new hospital]. AB - The "Azienda Ospedaliera Ospedali Riuniti di Bergamo" has completed the preliminary planning of the building of a new, great hospital, after a long and deep debate about the role and the significance of a modern hospital, thought to support, during the next century, the activity of a "hospital of national relevance and high specialization". The main aspects about the number of beds in the new hospital are debated and considered in the new conditions following the new programming acts of health planning in Italy and Lombardy and the needs of demography and emerging health demands The choice between new building or restructuring the existing hospital is discussed and the debate is referred to international scientific bibliography. Risk management is also considered as a must in a new hospital building. PMID- 12528336 TI - [Carbuncle in humans and animals: modern aspects of an ancient disease]. AB - Carbuncle, very remarkable disease in the past for its spreading, recently seems to be forgotten as risk for general population and exposed workers too, among them we remember especially veterinary physicians, breeders, tanners, etc. World epidemiological data aren't exact, but a lot of researchers in vary countries confirm that this is a present problem in many areas. Therefore it's indispensable to know correct diffusion of this disease, to fight it and other zoonosis in some environments and mostly in working ones, hoping phycians identify and declare new cases. PMID- 12528338 TI - [The hygiene-sanitary report]. AB - The main contents of a Hygienic-Sanitary report are listed below, once a few premises on the definition and objectives of hygiene, and on the normative pertaining Hygienic-Sanitary reports in the context of sanitary constructions planning are set. A hospital is a complex organism, and it must be planned as an "organized system" in which the various activities are disposed in the functional spaces according to a well-defined layout. In a Hygienic-Sanitary report, one should always remember that the correct approach to a hospital construction or restructuring project must be first urbanistic, with a precise diagram of fluxes and functions, and only then architectonic, with the consequent hospital typology. Different organizational models of the functional zones and of the relative traffic fluxes correspond in fact to different architectonical typologies. PMID- 12528339 TI - [Risk assessment as a tool for planning reconstruction or construction of hospitals]. AB - The risk assessment procedure enables the Hospital General Manager to set up and plan the measures and programs for the health protection of workers, patients and visitors. This crucial procedure on the preventive scenario could also provide useful informations about strategic choices to which the general manager is often recalled such those about old building restructuration or a new building construction. Specific models for transferring and elaborating data to this new field for applicating the results of risk assessment appear therefore to be necessary and the authors suggest a discussion about these theoretical models. PMID- 12528340 TI - [Diseases in hospital workers]. AB - Occupational diseases and labour accidents in health care workers (Azienda Ospedaliera di Padova) were evaluated from 1990 to 2000; the information collected is compared with the same data of Regione Veneto and INAIL. 262 occupational diseases were found; nurses are more affected than other health care workers. In both reports (Azienda Ospedaliera di Padova and Osservatorio epidemiologico della Regione Veneto) half of the occupational diseases and injuries are represented by allergic disease. Low back pain is also equally represented in Azienda Ospedaliera and in Osservatorio epidemiologico results (5.3% and 6.6% respectively). Among the labour accidents reported to INAIL from 1994 to 2000, 25% of them were represented by falls and slips, 23% by car accidents, 17% were caused by tools, 17% by movement of goods and persons, 8% by contact with splinters, liquids, dusts, 5% by assaults and 5% by contact with blood or body fluid. The latter result is widely underestimated in the INAIL report, because these events usually produce a temporary inability of less than 3 days; in fact, they represent more than half of the labour accidents treated at the Azienda Ospedaliera di Padova. Medical surveillance (in exposure to aldehydes, anaesthetic gases, chemioterapic drugs) has shown many other occupational health problems that often need specific structural intervention (restructuration and restoration of workplaces in health care environments), and work organization changes. PMID- 12528342 TI - [Ergonomics in planning and reconstruction]. AB - The state of well being of people is not represented only by any illness, but also by the satisfaction of all environmental components around him. For this reason, the hospitals demand new and more project attentions and in particular they demand specific attentions for all the environmental qualities (light, colours, indoor air qualities, and temperature), the equipment, the furniture and the privacy. This new attention to the requirements of people, during the phase of planning, needs the ergonomic studies. This kind of discipline, in fact, has got the objective of good relationships between men and context where they live. The study of ergonomic qualities needs different competence of multiple areas of acquaintance, that collaborate to analyse the requirements of sanitary structure, and in particular they analyse needs of different categories of people that work or stay in this place (the patients, visitors and sanitary staff). Moreover ergonomic studies concurs to analyse the various factors of complexity, of the buildings, that are very important for the project of hospital. From the scientific literature, we say that some physical environmental characteristics (light, colour, noise,) can influence the psychological conditions of people, and in particular they can generate three different categories of reactions: behavioural, neuropsychological, neuroendocrinal. For this reason the study of all the environmental factors, simple and composed, is a primary necessity in the hospital planning. In this way we can guarantee to the patient, a better sanitary service and a better comfort, and we can guarantee to all sanitary staff adequate conditions of security and functionality. PMID- 12528341 TI - [Planning and building a new hospital: how to reach adequate preventive standards]. AB - Communication deals with the complexity of planning a new hospital. We considered basic requirements needed to build this kind of settlement according to regulations in force and standards, with particular care to patients and workers safety and health, both during normal activity periods and in case of ordinary and extraordinary maintenance. PMID- 12528343 TI - [Hospital maintenance: management, risks, and responsibilities]. AB - Principal activities of maintenance carried out in hospital, staff required, various type of organization (inside team or global service), management aspects are described. Subjects responsible of maintenance are characterized and the relationships between Service of Maintenance and Service of Prevention and Protection in hospital are specified. Responsibility aspects concerning safety of maintenance workers and main risks which are exposed are defined. The importance of disponibility of a good maintenance handbook and of projects and programs of practice for new hospitals are emphasized. PMID- 12528344 TI - [New operating rooms: problems related with ventilation and air quality]. AB - Realizing the project of the new operating theatres, We bring out the importance of creating bright environments, rational and wide ways, a good indoor air quality, thermal comfort and occupational hygiene. Therefore for the first time We've installed an automatic system for permanent monitoring of gases, vapours and particles, associated with the study of ventilation, to evaluate the real condition of the mantenance and of staff behaviour. PMID- 12528345 TI - [Manual lifting of patients: experience at the Riuniti di Bergamo Hospital]. AB - The manual lifting of patients is a risk factor in the working activity of hospital nurses and it causes a high prevalence of low-back pain in these workers. The evaluation of risk has been performed in various Occupational Units of our hospital in accordance with art. 4 of Legislative Decree 626/94. The results of this evaluation, have led to implement the following preventive measures: health surveillance, professional training, purchase of special furniture and ergonomic supports. PMID- 12528346 TI - [Safety in planning]. AB - Our communication wants to stress the importance of choices which must be carried out during the planning stage in order that building works (and especially building hospitals) are realized in a safe environment. It is particularly emphasized the relationship between the regulation in force in matter of public building contracts and that in matter of safety in building process, both in case of new buildings and in case of ordinary and extraordinary maintenance. PMID- 12528347 TI - [Health surveillance in construction]. AB - In the Italian and international literature there are very few studies about work diseases and their prevention in the construction's compartment. The analysis of the most recent articles about this theme, however, suggests the presence of an high professional risk in the compartment and indicates as the most relevant risk factors noise, dusts, fibres, manual lifting, prolonged fixed postures, cumulative trauma disorders, vibrations, chemical substances and climatic conditions. The role that extra-lavorative risk factors can play on health is also relevant; those risks are due to the workers' peculiar life-style: hyper caloric diet, smoke and use of alcohol. Interventions of medical hygienic prevention, formation and information are therefore necessary. Lombardy Region, which from many years is careful to the problems about the prevention in building yards, has created a Working party who has elaborated the "Guide Lines for the sanitary surveillance in the building industry". The indications reported in the Guide Lines could represent, even for the occupational-doctors of the building contractors of the New Hospital, a good base for the working out of the workers' health care activity, in the respect of the dues of the law in force. PMID- 12528348 TI - [Theory of training and quality of safety in health]. AB - This article aims to define the concept of security, particularly in relation to concepts of risk, error, danger and adverse events. The authors go on to analyse the latest theory of adult training, applying it to healthcare safety whilst recognising that this is a complex facility. By using tools such as guidelines and risk management, the authors propose a new kind of corporate culture that enables certain initiatives, such as clinical governance and therefore learning organisation, to develop. This strategic organisation the company allows vital knowledge and experience, which lead to problem-solving and an increased guarantee of security for both internal and external clients, to become the common knowledge and property of the company as a whole and of all who operate in and around it. PMID- 12528349 TI - [Authorization and accreditation: management of the conversion of hospital buildings]. AB - The Emilia Romagna Region decided own accreditation and procedures pattern following decisions, by different levels and graduated stages: authorization, institutional accreditation and finally contractual agreements. For Emilia Romagna Health Administration, authorization step represented "the necessary minimum level of security to exercise health activity", carried out by self evaluation of general and specific requirements of health structures, outlined by specific check-list, to bring about the maximum time allowed five years realization of structure adjustment plans. Beginning from this context the Local Health Unit of Modena, introduced by an audit test of conformity levels caught up, the guide line of computer management of technical assistance, change to use or amplification of health permission of structure and installations take a census by the business administration. The method allows by operative guide lines standardization to certify the validity of innovations realized, in all Departements/Services, for own conformity of structures, plant design and installation, sanitary regulations and security for wards services, auxiliary rooms. The development of this method allow to link activity and professionality for different duties of supervision and inspection, to guarantee the maintenance of levels assurance of health structures. PMID- 12528350 TI - [Injuries with biological risk in a major hospital: analysis of a caseload]. AB - This article report preliminary data on injuries involving risks from biological agents occurred in healthcare workers of a major hospital over a 5-years period ('96-'00). The role of Occupational Competent Physician is underlined. Injuries' database, archives of occupational diseases, discharge diagnosis and notification of transmissible infectious diseases were consulted. 200 injuries (3/4 percutaneous) were on average registered yearly, with a decreasing trend. Subjects had a mean age of 34 years and 7 years of length of employment at the hospital. 60% of exposures occurred in nurses and 43% workers belonged to surgical areas. Serological data of the "source patients" were available for 1/3 of the cases and 35% of them were bloodborne viruses infected; 1 seroconversion for HCV was registered. On the whole, our results are similar to data from national and international literature. However a few factors limited the data collection and interpretation, mainly the lack of computer formats and different ways of coding and filing the information. The periodical analysis of injuries is useful to the Occupational Physician, particularly for educational intervention and sanitary surveillance of workers. Moreover, the Occupational Physician plays an important role in injuries prevention, especially in spreading the information on Standard Precautions, evaluating the human factor, reducing the "under reporting" of accidents, counselling, evaluating prognosis and resuming work. PMID- 12528351 TI - [Occupational injuries with biological risk at a hospital]. AB - A ten years retrospective study was carried out among the personnel of Padua Hospital to determine the rate of occupational exposure to blood and body fluids. 4738 injuries occurred between 1990 and 1999: 78% were percutaneous injuries 22% blood contacts. No case of operating room transmission of blood-borne infection was documented but it is clear that the risk is not zero and the number of injuries indicate the need for further training and reinforcement of universal precautions to reduce occupational exposures. PMID- 12528353 TI - [Occupational exposure to electromagnetic fields in physiotherapy departments]. AB - An assessment of the electromagnetic fields emitted from short and ultrashort wave diathermy sources and from magneto therapy operating at a frequency of 50 Hz was made to evaluate the occupational exposure to the operators. Operators were exposed to electromagnetic fields which rarely exceed the recommended limits of International Commission on Non-Ionizing Radiation Protection (ICNIRP) and American Conference of Governmental Industrial Hygienists (ACGIH). Only a measurement of electromagnetic fields near short wave diathermy equipment operating at a frequency of 27.12 MHz exposed operators to levels above those recommended by ICNIRP at a distance of 1 metre. Magnetic fields of magneto therapy exceeded residential exposition of 1 microtesla, at a distance of 4 metres. PMID- 12528352 TI - [Protein content of latex gloves used in hospitals]. AB - In this study the total protein content and the latex allergenic protein content in different types of medical gloves commonly used in our hospital were evaluated. The correlation between these two parameters and the correctness of the data provided by the glove manufacturers have been also verified. The main purpose of this study was to acquire information useful for preventing latex allergy in our hospital personnel. 29 different types of medical gloves were examined. The total protein contents were evaluated with Lowry modified method and the allergenic latex protein contents were evaluated with RAST-inhibition assay, both on glove samples and glove extracts. The correlation between inhibition percentages found in glove samples and in glove extracts and the correlation between total protein content and inhibition percentages determined in samples of the same gloves was verified. The highest concentrations of total proteins and allergenic latex proteins were found in examination powdered latex gloves and in surgical powdered latex gloves. A good correlation was observed between the total protein content and the inhibition percentages determined in samples of the same gloves, and between inhibition percentages found in glove samples and glove extracts. A significant amount of latex proteins was found in some brands of nitryl gloves. The clear association between the total protein levels and the allergenic latex protein levels suggests that the gloves with highest total protein content have the greatest allergenic potential. We believe that the total protein content is relevant from a preventive point of view for a proper gloves selection in the workplaces. Therefore, manufacturing companies should provide package inserts including at least the total protein contents and possibly allergenic latex protein levels, with specific measurement methods. Since the highest total protein and allergenic latex protein contents were found in latex powdered gloves, powder-free gloves must be of first choice. RAST inhibition assays directly on glove samples instead of glove extract seems to be a good reliable and faster alternative for the evaluation of the allergenic potential of latex gloves. For latex allergic subjects it is necessary to pay attention in choosing nitryl gloves, especially as an alternative to natural rubber gloves since some brands may contain significant amounts of allergenic latex proteins. PMID- 12528354 TI - [Occupational exposure to anesthetic gases at several hospitals]. AB - We considered data of samples collected in the period 1994-2001 in 83 operating rooms of 13 different public and private hospitals in Veneto Region. The anaestetic gases more used in operating rooms were nitrous oxide, isoflurane and, more recently, sevflurane. The mostly polluted positions were those in proximity of anaesthesiologic devices; the gases average concentrations were low in all hospitals examined, although some operating rooms demonstrated concentrations of anasthetic gases exceeding limit values. Lastly, the professional figures had different exposure to the various anaesthetic gases. PMID- 12528355 TI - High-density DNA arrays for comparative genomics and epidemiological studies in clinical microbiology. PMID- 12528356 TI - Scientists discover gene 'signature' for tumor's tendency to spread. PMID- 12528357 TI - FDA approves ViroSeq HIV-1 genotyping system. PMID- 12528358 TI - Response Biomedical releases positive preclinical results of RAMP cardiac marker tests. PMID- 12528359 TI - DNAPrint launches pan-genome screening platform ADMIXMAP. PMID- 12528360 TI - Molecular diagnostics: a healthcare perspective. AB - Recent progress in the development of molecular diagnostics in medicine has been rapid and the hope has been expressed by some people that it will soon be possible to have a detailed 'genetic readout' to assist in the diagnosis of treatment of a variety of diseases. However, such an outcome may be neither achievable nor desirable when viewed from the perspective of suppliers of healthcare. PMID- 12528361 TI - Single-nucleotide polymorphism detection using peptide nucleic acids. AB - Variations in the human DNA sequence between individuals can be an indication of predisposition to disease, affect the response to drug treatment, or more directly, be the fingerprint of an inheritable trait or defect. Significant efforts at improving the speed, accuracy and sensitivity of detecting such polymorphisms have led to the development of a number of powerful approaches. Sequence-specific base pairing between the strands of DNA, according to the Watson-Crick model, forms the basis of many detection systems. The crucial specificity of this hybridization reaction in discriminating between single base variations may be enhanced by using synthetic peptide nucleic acids as probes. The remarkable properties of these DNA analogs have been successfully exploited in several ways and the use of peptide nucleic acids has become an accepted addition to the collection of procedures available for genetic analysis. PMID- 12528362 TI - Single nucleotide polymorphism genotyping using locked nucleic acid (LNA). AB - Locked nucleic acid (LNA) is a new class of bicyclic high affinity DNA analogs. LNA-containing oligonucleotides confer significantly increased affinity against their complementary DNA targets, increased mismatch discrimination (delta Tm) and allow full control of the melting point of the hybridization reaction. LNA chemistry is completely compatible with the traditional DNA phosphoramidite chemistry and therefore LNA-DNA mixmer oligonucleotides can be designed with complete freedom for optimal performance. These properties render LNA oligonucleotides very well suited for SNP genotyping and have enabled several approaches for enzyme-independent SNP genotyping based on allele-specific hybridization. In addition, allele-specific PCR assays relying on enzymatically enhanced discrimination can be improved using LNA-modified oligonucleotides. The use of LNA transforms enzyme-independent genotyping approaches into experimentally simple, robust and cost-effective assays, which are highly suited for genotyping in clinical and industrial settings. PMID- 12528363 TI - Genomic medicine and neuroblastoma. AB - Neuroblastic tumors are a broad biological and clinical spectrum of neoplastic disease that has long captured the attention of clinicians and scientists alike. It is the most common solid extracranial tumor in children and accounts for 8-10% of all childhood tumors. Tumors are derived from neural crest cells and neural differentiation is common. Neuroblastoma is unique in that it presents with at least three distinct patterns of disease. Locoregional disease (Stage 1, 2, 3) does not metastasize to bone or bone marrow. Stage 4 is a systemic disease with widespread metastasis that responds to chemotherapy but many develop resistance. Stage 4s presents in infancy, is widespread and can spontaneously regress with no intervention, leaving a focus of fibrosis or calcification. Prognosis correlates with age, stage and tumor biological profile. The goal of this review is to provide an overview of the disease and highlight diagnostic, prognostic and therapeutic advances in neuroblastoma. Recommendations and resources for the evaluation and treatment of this disease are outlined. PMID- 12528364 TI - New approaches for biomarker discovery in lung cancer. AB - Lung cancer remains the most common cause of cancer death in the US and worldwide. Currently, there is no implemented population-based screening for lung cancer. Of all the markers identified, none have achieved sufficient diagnostic significance to reach clinical application. Here we discuss the status of lung cancer early diagnostics, and the genomic and proteomic approaches currently undertaken for biomarker discovery. We then introduce the ANTIBIOMIX approach that enables high-throughput target discovery by interrogating biological samples using a collection of thousands of polyclonal antibodies. The development of specific and sensitive diagnostic assays using patient's biological fluids, such as sputum and serum, will improve screening, monitoring of disease progression and treatment response, and surveillance for recurrence. PMID- 12528365 TI - Application of G-proteins in the molecular diagnosis of psychiatric disorders. AB - Mental disorders are highly prevalent and often difficult to diagnose. Although significant advances have been achieved in medication for mental disorders, the diagnosis and treatment monitoring of these disorders remain in a static situation. The absence of objective diagnostic 'gold standards', derives from the special complexity of diagnosis in psychiatry. Heterotrimeric G-proteins are crucial elements in post-receptor information transduction. These proteins have been implicated in the biochemical mechanism of action of drugs used to treat psychiatric disorders. G-protein measurements have unravelled a differential pattern characteristic of each of the major mental disorders. The accumulated data supports the potential use of G-protein measures as state-dependent markers for the biochemical diagnosis of mental disorders and as aid in the biochemical monitoring of the response to a specific treatment. PMID- 12528366 TI - Molecular diagnosis of Epstein-Barr virus. AB - Molecular techniques have become an important tool in Epstein-Barr virus diagnostics. In recent years novel real-time PCR formats and in situ techniques have been developed that offer increased time efficiency, reduced cross contamination, high reproducibility, high sensitivity and allow determination of viral loads. In the near future, widespread clinical application of these diagnostic modalities may provide increased knowledge of the pathophysiology of Epstein-Barr virus infection and may optimize treatment or even explore novel Epstein-Barr virus-related diseases. The monitoring of Epstein-Barr virus viral loads in different tissue compartments is currently being effectively used to assess the treatment response or prognosis in patients with oncological diseases or immunosuppression. This may also gain increasing importance in the nononcological environment. However, the general acceptance of molecular techniques will largely depend on improved standardization. PMID- 12528367 TI - Use of molecular techniques for the detection of antibiotic resistance in bacteria. AB - The introduction of antibiotics in the 1930s revolutionized the treatment of previously fatal infections. Unfortunately, bacteria adapted quickly to this threat and antibiotic resistance soon developed. Both penicillin and methicillin resistance in Staphylococcus aureus were noted almost immediately following the introduction of penicillin and the penicillinase-resistant penicillins respectively. Antibiotic susceptibility testing rapidly became an integral part of the management of patients with infectious disease and remains the basis for clinical decision-making and antibiotic selection. However, the fundamental principles underlying antibiotic susceptibility testing methods have remained largely unchanged over the last few decades. Due to the inherent time delay imposed by bacterial growth rates, culture-based systems have traditionally provided results several hours to days after initial isolation. Better understanding of the genetic basis of resistance has resulted in the development of molecular methods to detect the genetic changes or mutations present in resistant bacterial phenotypes. Microbiologists and clinicians are faced with increasing options for the use of genotypic or phenotypic methods. This review provides an insight into the current molecular methods available for the detection of antibiotic resistance and to provide a basis for evaluating the choice of testing method to be used. PMID- 12528368 TI - Detection of Shiga toxin-producing Escherichia coli in food. AB - Shiga toxin-producing Escherichia coli are emerging as a significant source of food-borne infectious disease all over the world. Illness caused by Shiga toxin producing E. coli can range from self limited, watery diarrhea to life threatening manifestations such as hemorrhagic colitis, hemolytic uremic syndrome or thrombotic thrombocytopenic purpura and death. Shiga toxin-producing E. coli can potentially enter the human food chain from a number of animal sources, most commonly by contamination of meat with feces or intestinal contents after slaughter or cross-contamination of unpasteurized milk products. Because of the low infectious dose of the O157:H7 Shiga toxin-producing E. coli strain, laboratory diagnosis of Shiga toxin-producing E. coli in food samples has developed a great importance. This review will focus on the microorganism, giving priority to illness prevention and Shiga toxin-producing E. coli detection in food. PMID- 12528369 TI - [Perceptual binding of sensory events: the hypothesis of inclusive characteristics]. AB - A solution to the problem of perceptual binding on a new conceptual basis is proposed. It introduces the concept of inclusive sensory characteristics, which is a response of a given perceptual level to the same sensory attributes or characteristics of a lower level, whose spatial organization or specific temporal succession constitutes an adaptively meaningful entirety. The sequence of inclusive characteristics forms a hierarchy: from sensory attributes to higher inclusive characteristics binding sensory data into unified images and scenes. Specific patterns of electric activity, which map inclusive characteristics, are relayed by back projections from upper to lower neuronal levels. This forms a cascade of top-down transfer of excitation, which stimulates neuronal structures whose signals correspond to the higher inclusive characteristic of a given act of perception. It is demonstrated that such mechanisms well agree with the psychological and neurophysiological experimental data and can ensure binding of sensory events at all perceptual levels. PMID- 12528370 TI - [Neurobiological problems of brain ischemia and post stroke epilepsy]. AB - A review. Current literature data concerning pathophysiological mechanisms of cerebral ischemia and poststroke epilepsy are presented. Results of investigations of structural-functional changes in activity of inhibitory inter neurons, mechanisms of brain plasticity, and main components of neuroprotection in ischemia are also discussed. The presented data confirm the current concept of apoptosis and necrosis as a consequence of injuring action of ischemia, the role of excitotoxicity in pathogenesis of ischemia and poststroke epilepsy, and different kinds of plasticity in recovery of brain functions. PMID- 12528371 TI - [Hemispheric asymmetry of visual evoked potentials during human recognition of facial emotional expressions]. AB - Visual evoked potentials (VEP) in standard 16 EEG derivations were recorded in 26 young men and 20 women during recognition of facial emotional expressions and geometric figures. The stimuli were presented on a computer screen in the center of the visual field or randomly in the right or left vision hemifields. Peak VEP latency and mean amplitude in 50-ms epochs were measured; spatiotemporal VEP dynamics was analyzed in a series of topographic maps. The right hemisphere was shown to be more important in processing emotional faces. The character of the asymmetry was dynamic: at earlier stages of emotion processing the electrical activity was higher in the right inferior temporal region compared to the left symmetrical site. Later on the activity was higher in the right frontal and central areas. The dynamic mapping of "face-selective" component N180 of VEPs revealed the onset of activation over the right frontal areas that was followed by the fast activation of symmetrical left zones. Notably, this dynamics didn't correlate with the hemifield of stimuli exposition. The degree of asymmetry was lower during presentation of figures, especially in the inferior temporal and frontal regions. The prominent asymmetry of information processes in the inferior temporal and frontal areas was suggested to be specific for recognition of facial expression. PMID- 12528372 TI - [Changes in characteristics of sensory evoked potentials in dynamics of monotonous activity]. AB - Visual and auditory evoked potentials were recorded in 19 volunteers during their monotonous activity. Three groups of subjects different in performance quality were set aside. Characteristics of evoked potential components were subjected to analysis of variance. The latency of P1 component of visual and auditory evoked potentials recorded in respective projection cortical areas significantly increased over the course of the long-term monotonous activity. These changes were associated with performance deterioration. The observed changes in performance quality and brain functional state were determined by individual features of subjects. PMID- 12528373 TI - [Long-term potentiation and unit evoked responses in the cingulate cortex of freely moving rats]. AB - Long-term potentiation (LTP) of synaptic efficacy is considered to be the most probable physiological mechanism of long-term memory. However, lack of understanding of cellular and subcellular mechanisms of LTP induction in freely behaving animals does not correspond to the importance of the problem. It was tested whether the characteristics of potentiation in the cingulate cortex after tetanization of the subiculocingulate tract (SCT) meet the criteria of true LTP (that passes all known stages in its development and lasts for more than a day in freely-behaving animals). Additionally, characteristics of spike responses to SCT stimulation and the effects of application of different glutamate receptor blockers were studied. Without application of GABA receptor blockers, the LTP lasted for more than 24 hours. Application of NMDA glutamate receptor blockers significantly inhibited field potentials evoke by testing stimulation. Short latency spike responses to SCT stimulation were recorded with low probability that increased with stimulation intensity. The obtained data reveal the possibility to compare the involvement of cingulate neurons in acquisition of adaptive behavior and changes in their spike responses during the LTP development in freely-moving rats. PMID- 12528374 TI - [Characteristic features of adaptation to experimental conditions in primates with various initial keeping conditions]. AB - Ethological study of emotional state and behavior was carried out in male macaques of two species (Macaca mulatta and Macaca fascicularis), which were divided in three groups according to their initial keeping: isolated, family, and gregarious. During instrumental conditioning, the initial keeping was changed for identical conditions (individual caging or sitting in a primatological chair). It was shown that adaptation to stress associated with a sharp enviromental change depended on initial keeping conditions. Monkeys brought up in family with experience of both intra- and interspecies interactions (for example, monkey- human) had the highest adaptation and learning abilities. Individual and gregarious keeping of monkeys impoverished their individual experience, and their adaptation to experimental conditions required additional efforts. The learning of individually caged monkeys was successful, if their keeping conditions were not changed. Initially gregarious monkeys were successfully trained after their habituation to contact with humans. PMID- 12528375 TI - [Activity of neurons in the pedunculopontine nucleus in conditioned instrumental appetitive reflex]. AB - Activity of 91 neurons in the compact and diffuse parts of the pedunculopontine tegmental nucleus (PPTg) was recorded in freely moving rabbits during execution of instrumental appetitive conditioning. Of the recorded neurons, 37.4% changed their activity in response to a conditioned stimulus, which is the evidence for the PPTg involvement in instrumental conditioning. Excitatory conditioned neuronal responses to the conditioned stimulus and food reinforcement significantly prevailed over inhibitory reactions. Neuronal responses to the conditioned stimulus were classified in several basic patterns reflecting stimulus effects, structure of the behavioral act, and reinforcement properties. These reactions indicate the involvement of the PPTg in attention, motor learning, and reinforcement. The revealed differences in associative reactive properties of the compact and diffuse parts of the PPTg to the conditioned stimulus and reinforcement point to the functional heterogeneity of this structure and suggest the leading role of the cholinergic part of the PPTg in instrumental appetitive conditioning and reinforcement as well as the essential contribution of the diffuse part into classical appetitive conditioning. PMID- 12528376 TI - [Dependence of different spike sequences on the mean neuronal firing rate in the neocortex in vivo and in vitro]. AB - Neuronal spikes were recorded extracellularly in rabbit visual cortex in vivo (88 cells) and in surviving slices of guinea pig sensorimotor cortex in vitro (50 cells). Spike sequences (SS) with monotonically increasing (SS+) and decreasing (SS-) interspike intervals were detected. Relative number of spikes of SS in the recording was closely associated with SS generation. The relative number of spikes was plotted against the average firing rate, this function had a biphasic character with the critical point around 7 Hz. The rate of change in interspike duration (the slope) was virtually independent of the firing rate, but was significantly different in vivo and in vitro conditions for both SS+ (325 and 180 ms/s, respectively) and SS- (270 and 160 ms/s, respectively). By and large, in vivo and in vitro the spike sequence parameters depended in the average firing rate in the same manner. The role of the spike sequences in rhythmic and information processes in neocortex is discussed. PMID- 12528377 TI - [Reorganization of the pattern of the brain somatosensory evoked potentials after spinal cord injury]. AB - Recordings of somatosensory evoked potentials (SEPs) and SEP brain maps of 23 patients with vertebral injury were analyzed. All patients were operated at the Clinic of Military Traumatology and Orthopedics, Military Medical Academy. SEPs (19 sites) were recorded in real time and mapped with "Brain surveyor" neuromapper. Brain lesions were localized before surgery by of MR imaging and CT standard techniques. The results demonstrate that electrical stimulation of peripheral nerves produces a substantially variable SEP pattern. It was shown that SEP formation in more than 60% of recordings suggests a favorable prognosis for rehabilitation, whereas if more than 50% of recording sites are unresponsive, the rehabilitation prognosis is unfavorable. The amplitude of early components (below 60 ms) increases, and that of the late waves reduces. PMID- 12528378 TI - [Depression-like changes in behavior and c-fos gene expression in dopaminergic brain structures in WAG/Rij rats]. AB - In WAG/Rij rats with genetic absence epilepsy, inborn changes in behavior were observed such as decreased level of locomotion, exploratory activity, and grooming reactions in the open-field test, increased immobility in the forced swimming test, and decreased sucrose consumption (anhedonia) as compared to Wistar rats completely lacking in seizure pathology. These behavioral alterations in WAG/Rij rats resemble the symptoms of human depression (psychomotor retardation, depressed mood, and anhedonia). No significant behavioral changes were found in the light-dark choice, social interaction, and elevated plus-maze tests. This suggests the absence of increased anxiety in WAG/Rij rats. In contrast to Wistar, WAG/Rij rats were sensitive only to chronic treatment with antidepressant imipramine like depressive patients. Behavioral "despair" induced by forced swimming led to C-fos gene expression in three brain structures (frontal cortex, nucleus accumbens, and striatum), which are, respectively, terminal regions of three dopaminergic brain systems (mesocortical, mesolimbc, and nigrostriatal). c-fos gene expression in the brain of WAG/Rij rats was substantially different from that in the brain of Wistar rats in both intensity (in WAG/Rij the c-fos gene expression was higher than in Wistar rats in all involved brain structures) and its distribution between the structures. The results suggest that WAG/Rij strain is a new experimental (genetic) model of absence epilepsy-related depression unassociated with increased anxiety. PMID- 12528379 TI - [Behavioral consequences of isolation in early ontogeny in rats: selectivity of anxiety conditions]. AB - Multiparameter scale for evaluation of anxiety-phobic state in rats reveals significant enhancement of anxiety in rat pups after 6-week isolation (beginning from the 21st day from birth) as compared to grouped controls of the same litter: the locomotion and exploration that appear in test areas are suppressed, and species-specific fear reactions are enhanced. These changes considered as signs of situational anxiety are not eliminated by 2.5-month keeping in groups. Nevertheless, they are not correlated with parameters of the acoustic startle reflex that (by the data of literature) is thought to be related with fear and anxiety. On the basis of the discrepancy it is proposed that state of anxiety is selective. This suggestion is confirmed by individual behavioral variations characterized by a combination of a low level of situational anxiety and a high level of acoustic anxiety observed in both experimental and control groups. These variations may explain the existence of atypical "emotional resonance"-like behavior according to P.V. Simonov. Attention is given to selectively enhanced acoustic startle reflex in the group of active control as an evidence for critical importance of any manipulations with social context in early ontogeny. PMID- 12528380 TI - [Influence of neurotensin on behavior of rats with lesions in serotonergic neurons]. AB - After serotonergic lesion by administration of 5,7-dihydroxytryptamine into the dorsalis raphe nucleus, effects of neurotensin microinjections into the caudate nucleus and substantia nigra on rat behavior were compared. Serotonergic lesions resulted in motivated excitement of rats manifested as an increase in the number of intersignal motor reactions during realization and, particularly, extinction of thirst conditioned reflex. Neurotensin microinjections into the caudate nucleus facilitated extinction of the conditioned reflex both in operated and control rats, but such microinjection into the substantia nigra facilitated this process only in operated animals. Neurotensin did not change conditioned reflex realization in both groups of animals but decreased emotional excitement of rats in the "open field". The behavioral effects of neurotensin in operated rats are connected with normalization of motivational and emotional states of animals and may be explained by recovery of interaction between the dopamine- and serotonergic systems. It is suggested that the mechanisms of this normalizing effects of neurotensin at the levels of the caudate nucleus and substantia nigra are different and are associated preferentially with its action either on dopamine- or serotonergic structures. PMID- 12528382 TI - [A method of minimization of ECG artifacts in digital EEG]. AB - An off-line algorithm of automatic detection and minimization of ECG artifacts in digital EEG without using the ECG channel is proposed. A possibility of application of this technique for processing another high-frequency artifacts and different waveforms in biosignals is discussed. PMID- 12528381 TI - [Regulation of posttetanic increase in choline sensitivity in Helix neurons by Na,K-pump: the role of Na/Ca-exchange and of mobilized calcium]. AB - We studied the role of Na/Ca-exchange and intracellular mobilized calcium in ouabain-mediated suppression of potentiation of cholinosensitivity of somatic membrane in Helix LPa3 and RPa3 command neurons of defensive behaviour after electrical orthodromic tetanisation of n. intestinalis. Cholinosensitivity of neurons was assessed by the amplitude of the inward current evoked by acetylcholine. Inhibitor of a Na/Ca-exchange benzamil and specific inhibitor of Ca-ATPase in endoplasmic reticulum thapsigargin prevented the development of the posttetanic potentiation (PTP). PTP did not arise and at joint action of ouabain with benzamil or thapsigargin. It was concluded that Na/Ca-exchange and mobilized calcium are involved in development of PTP of cholinosensitivity in somatic neuronal membrane and its regulation by Na,K-pump. PMID- 12528383 TI - [Simulation of local cerebral hemorrhage in different brain structures of experimental animals]. AB - A standardized experimental model of intracerebral hemorrhagic stroke in small laboratory animals is developed and advanced for chronic neurobiological studies of normal and pathological higher nervous activity as well as disorders developed after acute hemorrhages. A device is advanced which allows a researcher to destroy appropriate brain structures (tissues and local blood vessels) with necessary precision by four-six rotations of curved stereotaxically inserted mandrel-wire knife and, subsequently, to inject autoblood into the area of the lesion. The advanced model is convenient for the reproduction of lesions in different brain regions (for the purpose of experimental knockouts) in neurophysiological, neuropharmacological, and clinical investigations. PMID- 12528384 TI - The role of socioeconomic circumstances in differences in height of pre-school children within and between the Czech Republic and southern Brazil. AB - Children's height is an important indicator of nutritional status and health of populations. Little is known about the role of individual socioeconomic factors and whether socioeconomic differences within countries can help explaining differences in children's height between countries. This paper examines the effect of socioeconomic factors on children's height in two population with different social environments. Two separate cross-sectional studies of children 36-59 months old from Southern Brazil and the Czech Republic collected data on height and socioeconomic and demographic variables. Height was converted into height-for-age Z-scores (HAZ) based on one growth standard, and the effect of socioeconomic and demographic factors on children's heights was studied by multiple regression. First, these effects were estimated separately for each country, and secondly, it was assessed whether some of the difference in height between the two countries could be explained by these factors. Czech children were 0.61 HAZ (approximately 2.6 cm) taller than Brazilian children. In Brazil, income, dwelling conditions, birth order, birth weight and maternal age were independently and significantly associated with height (multiple R-squared 0.27). In the Czech Republic, maternal education, birth order, birth weight and maternal age predicted height in a multivariate analysis (multiple R-squared 0.11). Maternal education alone explained 30% of the difference in children's height between Brazil and the Czech Republic; further adjustment for education of the father, birth weight, number of siblings and birth order slightly increased the proportion of explained difference to 41%. Our results suggest that although socioeconomic and demographic factors predicted children's growth in both countries, the role of individual factors differed and the associations were stronger in South Brazil. The finding that these factors explained a large part of the between-population difference further illustrates the importance of social environment for children's growth. PMID- 12528386 TI - WHO launches the first global strategy on traditional and alternative medicine. PMID- 12528385 TI - Tobacco-related mortality following the Peto-Lopez epidemiological model used in international public health comparison. AB - In estimating the overall scale of death from tobacco in developed countries, there is a range of uncertainty between the lower limits and the upper limits of what is scientifically plausible. Following a method by Peto, Lopez et al. to base the calculations chiefly on the lower limits, we have compared the tobacco related mortality for females and males separately in various ages groups between Poland and Sweden over a period of three decades. This method is built on the following steps: ignore deaths under age 35, ignore cirrhosis and non-medical causes, compare lung cancer at ages 35-79 with US non-smokers and concerning other diseases at ages 35-79 make a conservative halving of apparent excess attributed to tobacco. Concerning mortality all causes a tobacco-related mortality among Polish men has increased more than 50% from the 5-year period 1965-1969 to the period 1990-1994. The analyse for women gives a strongly other picture compared with men showing much lower rates. When comparing the middle age group with the overall population it is interesting to find a faster increase in tobacco-related mortality rates for Polish men and for Swedish women in the age group 45-64 years than in the overall group. Swedish men have reduced their smoking habit strongly and far more than Polish men (17 and 42% respectively 1999). Concerning the pattern of tobacco use it is obvious that Swedish and Polish women during the last descades have had the same and slowly decreasing smoking prevalence (21 and 23% respectively 1999). It is important to use not only lung cancer mortality but a total picture of tobacco-related disorders as indicators of the success in tobacco control in a country as well for women as for men, especially in the middle age groups. PMID- 12528387 TI - Cost effectiveness and cost benefit of viral hepatitis B vaccination in the Slovak Republic. AB - In our work we tried to evaluate the cost effectiveness (CEA) and cost benefit (CBA) of the vaccination strategy of viral hepatitis B (VHB) vaccination in Slovakia. Retrospectively we analysed the incidence of VHB before and after the vaccination against VHB. From the calculated yearly cost (direct and indirect)- treatment costs and vaccination costs--we tried to calculate the estimated financial costs of vaccination strategy in 2000 (89.4 mil. SK), the saved costs (92 mil. SK), cost effectiveness (201 642 SK) and cost benefit (2.70 SK). First a decrease of the incidence (after introduction of measures to prevent nosocomial transmission) had been achieved by vaccination of selected risk groups of the population. A significant positive impact on the incidence of VHB is expected from mass preventive vaccination of infants, introduced in Slovakia in 1998. PMID- 12528388 TI - The effect of heat exposure on cortisol and catecholamine excretion rates in workers in glass manufacturing unit. AB - The aim of the investigation was to study the effect of long term repeated heat exposure on the excretion rates of stress hormones of workers in glass manufacturing unit. Sixteen operators, exposed to heat, were studied during the hot period and compared to a control group of 16 subjects, working in the same manufacturing unit. Both groups had moderate work load. The microclimate components and the Wet Bulb Globe Temperature were used for defining the heat exposure. The excretion rates of cortisol, adrenaline and noradrenaline were followed during the early morning shifts on three hour intervals using RIA and fluoriphotometric methods. Heart rate was followed, too. The psychosocial factors were measured by the "My job" questionnaire. Highly significantly higher cortisol, noradrenaline and adrenaline values were measured in the heat exposed operators compared to the control group, while significant differences of the psychosocial factors between the two groups lacked. Even if the heart rate was in the safe limits, the found alterations in the stress system are considerable and indicate heat stress. The work in conditions of overheat is associated with considerable heat stress and the possible health implications need to be clarified. PMID- 12528389 TI - Infectious complications in 135 Turkish renal transplant patients. AB - Infections are the emerging causes of mortality and morbidity due to lifelong immunosuppressive therapy in renal transplant patients (1, 4). Here, we report infectious complications of 135 renal allograft recipients who were followed up in the last 20 years in Gulhane Military Medical Academy, Ankara, Turkey. Of them, 83 (61.4%) had a transplant from living related donors, 18 (13.3%) from living non-related HLA matched donors and 34 (25.1%) from cadaveric matched donors. Immunosuppression was achieved in 42 (31.1%) recipients by azathioprine plus corticosteroid (AZA + CS) and in 93 (68.8%) by AZA + CS + cyclosporin A (CsA). Encountered infections were classified according to three different periods of the transplantation procedure [early (first month), intermediate (2 6th months) and late (after the 6th month)]. Bacterial infections were the leading infections in all three periods and the most affected system was the urinary tract. Each recipient had at least one episode of urinary tract infection (UTI) and E. coli was the most common urinary pathogen. On the other hand, HCV was the leading viral pathogen (14.3%). The total mortality rate was 7.4%, and septic shock was the most common cause of death (80%). PMID- 12528390 TI - Magnesium in drinking water and liver cancer morbidity--a possible relation? AB - The paper presents results of a research on liver cancer morbidity in Opole province, Poland, in relation to magnesium exposure in drinking water. Based on the extensive empirical materials of cancer registry information and water quality, the well known statistical approach using BUGS software was applied in the study. The results support a hypothesis of a possible association between the deficiency of magnesium in drinking water and the increase of liver cancer morbidity in the population exposed. The outcomes were presented in a table and graphically in histograms, scatterplots and maps. PMID- 12528391 TI - Gender-related differences in self-rating of health and functional status of elderly people with ischaemic heart disease. Comparative study in 12-year period. AB - PURPOSE OF THE STUDY: The study aim is to present changes in the self-rating of health and functional status in performing activities of daily living, reported by the elderly persons with ischaemic heart disease (IHD), at the time interval of 12 years. Gender-related differences as well as socio-medical characteristics have been involved in the analysis changes. DESIGN AND METHODS: The study was carried out in 607 elderly respondents who participated in two surveys at the time interval of 12 years. The analysis of the functional ability in performing daily activities in both surveys was based on the ADL and IADL scales, self-rated health was based on the scale ranging from very poor to excellent health. RESULTS: It has been observed in the baseline survey that the respondents with IHD reported lower mobility, functional activity and scored their health lower than other respondents. Though females with IHD scored higher their functional ability than males with the same disease, decrease of their functional ability at the time interval between both surveys was three-fold higher compared to males with the same disease. The results of the second study demonstrated age-related level of self-coping with the activities of daily living (ADL scale) in females with IHD, while none of the factors from the baseline survey had influence on their functional independence (irrespective of IHD). Functional status in the second study of males with IHD has been influenced by age and presented in baseline study willingness to life, while in males who have not reported IHD, any factor coming from the 1st survey have not determined the level of independence in activities of ADL in survey II. Functional ability in males with IHD was affected in the IInd study, besides age, by the attitude denying weariness of life demonstrated in the baseline survey. None of the factors from the baseline survey determined self-coping with daily activities by the males without IHD as it was observed in the IInd study. PMID- 12528392 TI - Bodily characteristics and lifestyle of Czech children aged 7.00 to 10.99 years, incidence of childhood obesity. AB - In 1991-2000 in the Czech Republic a survey of the prevalence of obesity was made. The assessment was made by the transversal method in elementary schools in all regions of the Czech Republic. Children aged 7.00 to 10.99 years were investigated. The survey comprised a total of 3362 children (1668 girls and 1694 boys). A total of 12 anthropometric dimensions were recorded and questionnaires addressed to the children and parents were analyzed. From the results ensued that the ratio of obese children of both sexes in the CR (i.e. children with BMI values about the 97th percentile of the reference population) increased. This zone comprised 6.0% boys and 5.6% girls of the investigated group. An important finding was also the increase of mean values of the abdominal circumference of the children in all age groups in both sexes. The ratio of obese children in different communities is related unequivocally to the size of the community., In smaller communities there are more obese children (according to our data 6.9%) and in large towns the ratio of obese children is 2.3%. In large towns there is also a greater proportion of thin children - 12.1%, in small communities 8.6%. It was found that there is a higher percentage of obese children in families where the mother has elementary education as compared with children of mothers with secondary and university education. Mothers with elementary education have 9.6% obese children while university educated mothers have only 3.6% obese children. Analysis of the dietary questionnaires revealed that only 62.9% children in large towns eat breakfast, the position in villages is similar - 63.9%. In the group of children with excessive body weight fewer children have breakfast (only 54.1%), in the group of children with a low body weigh the majority eat breakfast (75.3%). The majority of children have a mid-morning snack at school which they bring from home. As regards school lunch the majority of children from large towns replied that they have lunch at school. the number of rural children who had school lunch was smaller. An afternoon snack is taken by less than half the children in different groups. PMID- 12528393 TI - Safe and sound. Tackling the wandering problem with products and strategies. PMID- 12528394 TI - Simply strategy. It can be a real problem insuring long term care facilities--but not if you use some practical solutions. PMID- 12528395 TI - Postmortem procedures. Services for the deceased need the same respect as when the person was alive. PMID- 12528396 TI - When it's time to say 'good-bye'. PMID- 12528397 TI - Get 'em while they're young. The boomers can really impact senior living--but can we wait until 2021? PMID- 12528398 TI - In$ide information. Two new lender and investor surveys give insight into the financing environment. PMID- 12528399 TI - A new world. Handling diversity among staff and residents means going back to [everyone's] values. PMID- 12528400 TI - Skin signs of systemic disease: an update. PMID- 12528402 TI - Treatment of inflammatory dermatoses with novel biologic agents: a primer. PMID- 12528401 TI - Cytokines and other biologic agents as immunotherapeutics for cutaneous T-cell lymphoma. AB - Cutaneous T-cell lymphomas (CTCLs) are a group of skin-invasive malignancies of clonally derived T lymphocytes. Mycosis fungoides and Sezary syndrome, characterized by the proliferation of CD4+ T cells, are the most common forms of CTCL. Among these latter disorders, the host antitumor response appears to play an important role in disease control. Thus, systemic therapeutic agents are used in an effort to augment the host antitumor response while selectively targeting the malignant cells. Both new and old biologic response-modifying treatment options currently used to treat CTCL are reviewed. PMID- 12528403 TI - The cutaneous neurosensory system in skin disease. AB - The cutaneous neurosensory system appears to be involved in a number of skin diseases. Ongoing studies continue to uncover potential new roles for the components of the neurosensory system in skin homeostasis and disease states. There is new evidence that neuropeptides may play a role in melanogenesis with effects on vitiligo. An increase of intraepidermal nerve fibers with a possible pathophysiologic role in photodamaged facial skin has been proposed. As our understanding of the interactions between the cutaneous neurosensory system and the various components of the skin and the immune system in times of health and disease increases, specific treatments modulating the neurocutaneous system will find their way into the armamentarium of daily dermatologic therapy. PMID- 12528404 TI - Molecular genetics of the inherited disorders of cornification: an update. PMID- 12528405 TI - Stevens-Johnson syndrome and toxic epidermal necrolysis. PMID- 12528406 TI - Epidermal nevi. PMID- 12528407 TI - Recent advances in intraepidermal blistering diseases. PMID- 12528408 TI - Effects of ultraviolet irradiation on inflammation in the skin. PMID- 12528409 TI - Apoptosis and cutaneous biology. PMID- 12528410 TI - Advances in wound closure materials. PMID- 12528411 TI - Actinic keratoses: new treatment options. PMID- 12528413 TI - Clinical negligence claims: specialist practitioners and legal aid--and the new rules. PMID- 12528412 TI - Access to justice--let them eat cake. PMID- 12528414 TI - The impact of the new civil practice rules on clinical negligence claims. PMID- 12528415 TI - Removing the moral sting from medical malpractice litigation: a modest proposal based on the Canadian experience. PMID- 12528416 TI - The doctor and the lawyer: medico-legal problems today. PMID- 12528417 TI - Recovered memories: some aspects of the controversy. PMID- 12528418 TI - Human rights in the new millennium. PMID- 12528419 TI - Nurse vacancy rate. PMID- 12528420 TI - Revisioning the nursing shortage: a call to caring for healing the healthcare system. AB - The nurse shortage is one of the most serious threats to quality care facing the healthcare system. This article examines the situation through multiple lenses and suggests that an ecological model of resource conservation and the creation of habitats for healing might be better solutions than traditional economic and business frameworks can offer. These habitats for healing would be the central elements of a renewed, healing healthcare system, allowing the reemergence of the caring and healing environment necessary for attracting and keeping more nurses. PMID- 12528421 TI - Why nurses stay. PMID- 12528422 TI - The nursing shortage: insights and perceptions. PMID- 12528423 TI - New models of nursing care. PMID- 12528424 TI - Child emotional security and interparental conflict. AB - Guided by the emotional security hypothesis developed by Davies & Cummings (1994), studies were conducted to test a conceptual refinement of children's adjustment to parental conflict in relation to hypotheses of other prominent theories. Study 1 examined whether the pattern of child responses to simulations of adult conflict tactics and topics was consistent with the emotional security hypothesis and social learning theory in a sample of 327 Welsh children. Supporting the emotional security hypothesis, child reports of fear, avoidance, and involvement were especially prominent responses to destructive conflict. Study 2 examined the relative roles of child emotional insecurity and social cognitive appraisals in accounting for associations between parental conflict and child psychological symptoms in a sample of 285 Welsh children and parents. Findings indicated that child emotional insecurity was a robust intervening process in the prospective links between parental conflict and child maladjustment even when intervening processes proposed in the social-cognitive models were included in the analyses. Studies 3 and 4 explored pathways among parental conflict, child emotional insecurity, and psychological adjustment in the broader family context with a sample of 174 children and mothers. Supporting the emotional security hypothesis, Study 3 findings indicated that child insecurity continued to mediate the link between parental conflict and child maladjustment even after specifying the effects of other parenting processes. Parenting difficulties accompanying interparental conflict were related to child maladjustment through their association with insecure parent-child attachment. In support of the emotional security hypothesis, Study 4 findings indicated that family instability, parenting difficulties, and parent-child attachment insecurity potentiated mediational pathways among parental conflict, child insecurity, and maladjustment. Family cohesiveness, interparental satisfaction, and interparental expressiveness appeared to be protective factors in these mediational paths. No support was found for the social learning theory prediction that parent-child warmth would amplify associations between parental conflict and child disruptive behaviors. PMID- 12528425 TI - Mechanisms in the development of emotional organization. PMID- 12528426 TI - Why a "word order difference" is not always a "word order" difference: a reply to Weyerts, Penke, Munte, Heinze, and Clahsen. AB - We present evidence that the supposed processing advantage for an SVfinO word order over an SOVfin word order in German argued for by Weyerts, Penke, Munte, Heinze, and Clahsen (2002) is supported by neither experimental nor theoretical evidence. Specifically, we show (a) that the frontocentral negativity for an SOVfin in comparison to an SVfinO word order in Weyerts et al.'s Experiments 2 and 3 is reducible to more general differences in the electrophysiological responses elicited by nouns versus verbs in a sentence context, and (b) that the P600 difference between the two word orders in Experiment 2, as well as the reading time differences in Experiment 1, result from the fact that the two supposedly ungrammatical conditions actually differ in their degree of ill formedness. We conclude that there is no evidence for a processing disadvantage for SOVfin, thus reconciling Weyerts et al.'s results on German sentence processing with the grammatical regularities of German. PMID- 12528427 TI - The role of language in memory for actions. AB - Languages differ with respect to how aspects of motion events tend to be lexicalized. English typically conflates MOTION with MANNER, but Japanese and Spanish typically do not. We report a set of experiments that assessed the effect of this cross-linguistic difference on participants' decisions in a similarity judgment task about scenes containing novel animations as stimuli. In Experiment 1, which required participants to encode the stimuli briefly into memory, we observed a language effect; in Experiment 2, which required participants to analyze the same stimuli, but not remember them, the language effect disappeared. Hence, these experiments reveal a task-dependent effect, which, we argue, points to working memory as the source of the language effect observed in Experiment 1 and, potentially, other experiments that have shown a linguistic relativity effect. PMID- 12528428 TI - The nature of regularity and irregularity: evidence from Hebrew nominal inflection. AB - Most evidence for the role of regular inflection as a default operation comes from languages that confound the morphological properties of regular and irregular forms with their phonological characteristics. For instance, regular plurals tend to faithfully preserve the base's phonology (e.g., rat-rats), whereas irregular nouns tend to alter it (e.g., mouse-mice). The distinction between regular and irregular inflection may thus be an epiphenomenon of phonological faithfulness. In Hebrew noun inflection, however, morphological regularity and phonological faithfulness can be distinguished: Nouns whose stems change in the plural may take either a regular or an irregular suffix, and nouns whose stems are preserved in the plural may take either a regular or an irregular suffix. We use this dissociation to examine two hallmarks of default inflection: its lack of dependence on analogies from similar regular nouns, and its application to nonroots such as names. We show that these hallmarks of regularity may be found whether or not the plural form preserves the stem faithfully: People apply the regular suffix to novel nouns that do not resemble existing nouns and to names that sound like irregular nouns, regardless of whether the stem is ordinarily preserved in the plural of that family of nouns. Moreover, when they pluralize names (e.g., the Barak-Barakim), they do not apply the stem changes that are found in their homophonous nouns (e.g., barak-brakim "lightning"), replicating an effect found in English and German. These findings show that the distinction between regular and irregular phenomena cannot be reduced to differences in the kinds of phonological changes associated with those phenomena in English. Instead, regularity and irregularity must be distinguished in terms of the kinds of mental computations that effect them: symbolic operations versus memorized idiosyncrasies. A corollary is that complex words are not generally dichotomizable as "regular" or "irregular"; different aspects of a word may be regular or irregular depending on whether they violate the rule for that aspect and hence must be stored in memory. PMID- 12528429 TI - On the categorization of aphasic typologies: the SOAP (a test of syntactic complexity). AB - This paper presents a new measure of syntactic comprehension abilities in brain damaged populations known as the SOAP (Subject-relative, Object-relative, Active, and Passive), along with data supporting its sensitivity and specificity. This assessment tool examines comprehension of sentences (matched for length) of four syntactic construction types: active, passive, subject-relative, and object relative. Data are presented that indicate that the SOAP provides a sensitive and reliable differentiation of aphasia subgroups. The SOAP's sensitivity in differentiating broad behavioral (anterior/posterior-lesioned) groups is compared to the auditory comprehension component of the Boston Diagnostic Aphasia Examination (BDAE), supporting its sensitivity differentiating between anterior- and posterior-lesioned groups. It is argued that this tool can be an important accompaniment to standard aphasia assessment batteries in more sensitively defining syntactic comprehension deficits. PMID- 12528430 TI - Antecedent priming at trace positions in Japanese long-distance scrambling. AB - We report the results from three cross-modal lexical decision experiments investigating antecedent priming effects in Japanese. In the first two experiments we examined antecedent reactivation at the preverbal trace position in long-distance scrambling sentences. We found an interaction between the participants' working memory (WM) span and antecedent priming. For the high span group, the magnitude of antecedent priming at the trace position was significantly larger than at the earlier control position; for the low span group, on the other hand, there was no such difference. In a third experiment, we examined whether similar reactivation effects could be observed for argument expressions that are not base-generated adjacent to the verb. Contrary to scrambled objects, subject noun phrases (NPs) in canonically ordered sentences were not reactivated at the preverbal test position in either of the two participant groups. We argue that the priming effect observed in the high span group supports a trace-based account of long-distance scrambling. The degree of complexity of the experimental sentences was such that they exceeded the memory span of the low span group. We conclude that argument traces access their antecedents irrespective of the position of their subcategorizers. PMID- 12528431 TI - Use of released pigs as sentinels for Mycobacterium bovis. AB - Identifying the presence of bovine tuberculosis (TB; Mycobacterium bovis) in wildlife is crucial in guiding management aimed at eradicating the disease from New Zealand. Unfortunately, surveys of the principal wildlife host, the introduced brushtail possum (Trichosurus vulpecula), require large samples (> 95% of the population) before they can provide reasonable confidence that the disease is absent. In this study, we tested the feasibility of using a more wide-ranging species, feral pig (Sus scrofa), as an alternative sentinel capable of indicating TB presence. In January 2000, 17 pigs in four groups were released into a forested area with a low density of possums in which TB was known to be present. The pigs were radiotracked at 2 wk intervals from February to October 2000, and some of them were killed and necropsied at various intervals after release. Of the 15 pigs successfully recovered and necropsied, one killed 2 mo after release had no gross lesions typical of TB, and the only other pig killed at that time had greatly enlarged mandibular lymph nodes. The remainder were killed at longer intervals after release and all had gross lesions typical of TB. Mycobacterium bovis was isolated from all 15 pigs by mycobacterial culture. Home range sizes of pigs varied widely and increased with the length of time the pigs were in the forest, with minimum convex polygon range-size estimates averaging 10.7 km2 (range 4.7-20.3 km2) for the pigs killed after 6 mo. A 6 km radius around the kill site of each pig would have encompassed 95% of all of their previous locations at which they could have become infected. However, one pig shifted 35 km, highlighting the main limitation of using unmarked feral pigs as sentinels. This trial indicates use of resident and/or released free-ranging pigs is a feasible alternative to direct prevalence surveys of possums for detecting TB presence. PMID- 12528432 TI - Epizootiologic studies of avian vacuolar myelinopathy in waterbirds. AB - Epizootic avian vacuolar myelinopathy (AVM) was first recognized as a neurologic disease in bald eagles (Haliaeetus leucocephalus) and American coots (Fulica americana) in Arkansas, USA in 1994 and 1996, respectively, but attempts to identify the etiology of the disease have been unsuccessful to date. Between 1998 and 2001, wing clipped sentinel birds (wild American coots and game farm mallards [Anas platyrhynchos]) were released at Lake Surf, North Carolina, a lake with recurrent outbreaks of AVM, in order to gain a better understanding of the epizootiology of the disease. As early as 5-7 days post-release, sentinel coots and mallards showed neurologic signs of disease and were confirmed with AVM upon histologic examination of their brains. Serial releases of sentinel mallards during the summer, fall, and winter of 2000-01 demonstrated that exposure to the causative agent at a threshold sufficient to manifest disease was seasonal and occurred over about a 2 mo period, during November and December. Our findings that disease onset can be very rapid (5-7 days) and that exposure to the causative agent of AVM is site-specific, seasonal (late fall to early winter), and occurs over a relatively short duration (several months) supports the hypothesis that the disease is caused by a chemical substance, most likely of natural origin. PMID- 12528433 TI - Morbidity and mortality factors in key deer (Odocoileus virginianus clavium). AB - The population health of endangered Key deer (Odocoileus virginianus clavium) was monitored from 10 February 1986 to 28 September 2000 by necropsy of animals that were killed by vehicles, euthanized because of terminal injuries or disease conditions, or found dead. The predominant mortality factor during the period was collision with motor vehicles; however, several infectious diseases were diagnosed, including infections with Arcanobacterium pyogenes, Haemonchus contortus, Salmonella spp., and Mycobacterium avium subsp. paratuberculosis. During the period monitored, the only infectious disease that was thought to have affected population dynamics was haemonchosis. Nevertheless, several of the observed diseases have potential to impact viability of the Key deer population under appropriate environmental conditions. PMID- 12528434 TI - Cryptorchidism in Florida panthers: prevalence, features, and influence of genetic restoration. AB - The overall prevalence of cryptorchidism in Florida panthers (Puma concolor coryi) from 1972-2001 was 49% (24/49), with a significant increase over time. The earliest age at which descent of both testicles was known to occur was 2 mo and the latest was 10-13 mo. Delayed testicular descent was documented in 23% (8/35) of juveniles examined. Most retained testicles were in the inguinal canal. There was no apparent difference in reproductive success between cryptorchid and normal panthers, although no bilaterally cryptorchid panthers were known to have sired litters. Cryptorchidism was thought to be a manifestation of inbreeding and was one of several factors that led to the development of a genetic restoration plan whereby eight female puma from Texas were released into the panther population in 1995. None of the progeny resulting from genetic restoration efforts has been cryptorchid. This report provides evidence that cryptorchidism in panthers is genetically rather than environmentally based, and demonstrates the utility of genetic restoration for eliminating certain deleterious traits that result from inbreeding. PMID- 12528436 TI - Mercury in bald eagle nestlings from South Carolina, USA. AB - Bald eagles (Haliaeetus leucocephalus) may be at risk from contaminants in their diet and young birds may be particularly sensitive to contaminant exposure. To evaluate potential risks from dietary mercury exposure to eagle nestlings in South Carolina (USA), we surveyed mercury concentrations in 34 nestlings over two breeding seasons (1998 and 1999). Samples were also obtained from several post fledging eagles in the region. Nestling feather mercury ranged from 0.61-6.67 micrograms Hg/g dry weight, nestling down mercury from 0.50-5.05 micrograms Hg/g dry weight, and nestling blood mercury from 0.02-0.25 microgram Hg/g wet weight. We did not detect significant differences in tissue mercury between nestlings from coastal and inland regions in contrast to some other studies of piscivorous birds. Mercury concentrations were much higher in the post fledging birds we sampled. Our data show that nestling eagles in South Carolina are accumulating mercury, and that concentrations in older birds may exceed regulatory guidelines. PMID- 12528435 TI - Morbidity and mortality of reptiles admitted to the Wildlife Center of Virginia, 1991 to 2000. AB - Medical records from 694 reptiles admitted to the Wildlife Center of Virginia (WCV; Waynesboro, Virginia, USA) from 1991 to 2000 were reviewed to determine causes of morbidity and mortality. Eighteen species were represented but the majority of cases were four species; eastern box turtle (Terrapene carolina) (66%), eastern painted turtle (Chrysemys picta) (11%), common snapping turtle (Chelydra serpentina) (10%), and rat snake (Elaphe sp.) (6%). There was a significant increase in reptile cases during the study period both in absolute number and in proportion to the total caseload. Trauma (74%) was the most frequent cause of morbidity and mortality followed by unknown or undetermined (13%), aural abscessation (7%), infectious diseases (2%), and one nutritional disorder (0.1%). In addition, 3% of the cases were healthy animals that had been removed from the wild and consequently brought to the WCV. Causes of morbidity and mortality differed between the four most numerous species. Impact with a motor vehicle was the most frequent cause of trauma for eastern box turtles, eastern painted turtles, and common snapping turtles; however, garden-equipment related trauma was the most frequent cause for rat snakes. Aural abscessation was only seen in eastern box turtles. Eighty percent of cases occurred between May and September and 65% occurred within the five counties closest to the WCV. The majority of morbidity and mortality was the result of human activities. The expanding human population in Virginia likely will continue to have an impact on the health of wild reptiles. PMID- 12528437 TI - Innate resistance to epizootic hemorrhagic disease in white-tailed deer. AB - Differences in innate disease resistance at the sub-species level have major implications for wildlife management. Two subspecies of white-tailed deer, Odocoileus virginianus borealis and O. virginianus texanus were infected with epizootic hemorrhagic disease (EHD) viruses. These viruses are highly virulent pathogens of white-tailed deer and are endemic within the range of O. virginianus texanus but not within the range of O. virginianus borealis. Two experimental infections were performed. Five O. virginianus texanus fawns and five O. virginianus borealis fawns were infected with 10(7.1) median tissue culture infective doses (TCID50) of EHD virus, serotype 1 and five of each subspecies were infected with 10(7.1) TCID50 of EHD virus, serotype 2. Infections with both EHD virus serotypes caused severe clinical disease and mortality in O. virginianus borealis fawns, whereas disease was mild or nondetectable in O. virginianus texanus fawns. Virus titers and humoral immune response were similar in both subspecies suggesting that differences in innate disease resistance explain the differences seen in clinical disease severity. In white-tailed deer, innate disease resistance may vary at the subspecies level. Should this phenomenon occur in other species, these findings have major implications for managing wildlife populations, both endangered and non-endangered, using tools such as translocation and captive propagation. PMID- 12528438 TI - Cross-protection between epizootic hemorrhagic disease virus serotypes 1 and 2 in white-tailed deer. AB - Viruses in the epizootic hemorrhagic disease (EHD) serogroup are the most frequent cause of hemorrhagic disease in the southeastern United States, but nothing is known about cross-protection between the two EHD serotypes (EHDV-1 and EHDV-2) present in this region. We experimentally tested whether deer surviving EHDV-2 infection would be protected against subsequent infection with EHDV-1, and used field data to examine the possibility of reciprocal cross-protection. Eleven white-tailed deer fawns (Odocoileus virginianus) were experimentally infected with EHDV-2 and later challenged with EHDV-1. Two EHDV-2-naive fawns also were infected with EHDV-1. Deer were monitored via physical examination, complete blood counts, clotting profiles, viral isolation, and serology, and each animal was assigned a quantitative clinical disease severity score based on presence of certain physical and clinical parameters. Infection of naive controls with EHDV-1 caused severe clinical disease and death of both fawns, whereas deer previously infected with EHDV-2 exhibited no or minimal signs of disease. Thus, infection with EHDV-2 conferred protection against disease caused by subsequent EHDV-1 infection. Although prior EHDV-2 exposure protected deer from severe clinical disease, it did not prevent infection nor viremia indicating they could still act as virus amplifying hosts. These experimental infections suggest that EHDV-1 and 2 may exist in a state of mutual permissiveness. PMID- 12528439 TI - Paratuberculosis in key deer (Odocoileus virginianus clavium). AB - Paratuberculosis was diagnosed in an endangered Key deer (Odocoileus virginianus clavium) in November 1996. Between 10 April 1997 and 28 September 2000, the Key deer population was monitored for infection with Mycobacterium avium subsp. paratuberculosis by necropsy of available carcasses (n = 170), fecal cultures, and serology. One additional clinically affected Key deer was discovered in July 1998, and M. avium subsp. paratuberculosis was cultured from the feces of one live, asymptomatic deer. The results of this study provided sufficient evidence to consider the Key deer herd infected with M. avium subsp. paratuberculosis at very low prevalence. PMID- 12528440 TI - Immune responses of bison to ballistic or hand vaccination with Brucella abortus strain RB51. AB - From January through July of 2000, a study was conducted to evaluate clearance, immunologic responses, and potential shedding of Brucella abortus strain RB51 (SRB51) following ballistic or subcutaneous (SQ) vaccination of 7 mo old bison (Bison bison) calves. Ten bison calves were vaccinated SQ with 1.4 x 10(10) colony-forming units (CFU) of SRB51 and five calves were inoculated SQ with sterile 0.15 M sodium chloride. An additional 10 bison calves were ballistically inoculated in the rear leg musculature with 1 x 10(10) CFU of SRB51 and five calves were ballistically inoculated with an empty Biobullet. Serologic responses were monitored at 0, 2, 4, 6, 8, 12, 18, and 24 wk using the standard tube agglutination test and a dot-blot assay. Swabs from rectal, vaginal, nasal, and ocular mucosal surfaces, and blood were obtained for culture from all bison at 2, 4, 6, and 8 weeks post-inoculation to evaluate potential shedding by vaccinated bison or persistent septicemia. The superficial cervical lymph node was biopsied in eight ballistic and eight hand vaccinated bison at 6 or 12 wk to evaluate clearance of the vaccine strain from lymphatic tissues. Lymphocyte proliferative responses to irradiated SRB51 bacteria were evaluated in peripheral blood mononuclear cells (PBMC) at 4, 6, 8, 12, 18, and 24 wk after inoculation. Serum obtained from hand or ballistically vaccinated bison demonstrated antibody responses on the dot-blot assay that were greater than control bison (saline or empty Biobullet) at 2, 4, 6, and 8 wk after vaccination. Antibody titers of ballistically vaccinated bison did not differ (P > 0.05) from hand vaccinated bison at any sampling time. Blood samples obtained from all bison at 2, 4, 6 and 8 wk after vaccination were negative for SRB51. One colony of SRB51 was recovered from the vaginal swab of one ballistically vaccinated bison at 2 wk after vaccination. All other ocular, vaginal, nasal, and rectal swabs were culture negative for SRB51. Strain RB51 was recovered from superficial cervical lymph nodes of hand and ballistic vaccinated bison at 6 (two of four and two of four bison, respectively) and 12 wk (three of four and one of four bison, respectively). Serologic tests and bacterial culture techniques failed to demonstrate infection of nonvaccinated bison. Peripheral blood mononuclear cells obtained from hand vaccinated bison had greater (P < 0.05) proliferative responses to strain RB51 bacteria when compared to PBMC from nonvaccinated and ballistically vaccinated bison. Proliferative responses of PBMC from ballistically vaccinated bison did not differ (P > 0.05) at any sampling time from proliferative responses of PBMC from control bison. Serum alpha 1-acid glycoprotein concentrations, plasma fibrinogen, and total protein concentrations were not influenced by treatments. Ballistic delivery of SRB51 did not induce adverse effects or influence clearance of the vaccine strain. There were no proliferative responses of PBMC to SRB51 in bison ballistically vaccinated with SRB51; whereas bison inoculated with SRB51 by hand injection had greater proliferative responses than control or ballistically vaccinated bison. Our study suggests that ballistic delivery may require a greater dose of SRB51 to induce cell-mediated immune responses in bison that are comparable to those induced by hand injection, and that ballistic or hand delivery of 1 x 10(10) CFU of SRB51 is safe in bison calves. PMID- 12528441 TI - Immune responses of elk to vaccination with Brucella abortus strain RB51. AB - In a study conducted from January to August 2000, elk (Cervus elaphus) were vaccinated with Brucella abortus strain RB51 (SRB51, n = 6) or injected with 0.15 M NaCl solution (n = 3) at approximately 6 mo of age. Beginning at 2 wk and continuing to 25 wk after vaccination, SRB51-vaccinated elk had greater antibody responses (P < 0.05) to SRB51 when compared to nonvaccinated elk. Peripheral blood mononuclear cells (PBMC) from SRB51-vaccinated elk had greater (P < 0.05) proliferative responses to SRB51 at 18 wk after vaccination when compared to responses of nonvaccinated elk. Strain RB51 was recovered from blood samples of all vaccinates at 2 wk, and three of six vaccinates at 4 wk after vaccination. The SRB51 vaccine strain was recovered from the superficial cervical lymph node of all vaccinates sampled at 6 wk after vaccination. but not from lymph node samples obtained from vaccinates at 12 or 18 wk after vaccination. At 34 wk after vaccination, SRB51 was recovered from the bronchial lymph node of one of five vaccinates but not from other tissues. Strain RB51 was not recovered at any time from samples obtained from nonvaccinated elk. This study suggests that following vaccination with SRB51, elk remain bacteremic for a prolonged period of time, rapidly develop high antibody titers, and are slower to develop detectable proliferative responses in PBMC when compared to responses of cattle or bison (Bison bison). PMID- 12528442 TI - An indirect ELISA to detect the serologic response of elk (Cervus elaphus nelsoni) inoculated with Brucella abortus strain RB51. AB - An indirect enzyme-linked immunosorbent assay (ELISA) was developed to identify elk (Cervus elaphus nelsoni) with Brucella abortus strain RB51 (RB51)-specific antibodies using a mouse monoclonal antibody specific for bovine IgG1. This test was relatively easy to perform, accurate, and easily reproducible; therefore it could be standardized for use between laboratories. In addition, we attempted to compensate for inherent variabilities encountered when comparing ELISA readings from multiple samples taken from many animals over time. Optical density (OD) readings for each sample were converted into a percent positivity value for analysis. A negative cutoff value was determined above which a sample was considered to have a significantly elevated anti-RB51 antibody level. Pre- and postvaccination sera from 64 6-8 mo old elk, divided into four groups (females subcutaneously inoculated with saline (control animals), females ballistically inoculated with RB51, females subcutaneously inoculated with RB51, and males subcutaneously inoculated with RB51) were used. All serum samples were collected between 27 April and 15 November 1995. Values for all saline controls were appropriately below the negative cutoff value. All subcutaneously and ballistically inoculated elk were serologically positive to RB51 for at least two sampling periods during the study. The difference in percent positivity values for the ballistically compared to the subcutaneously inoculated groups was not statistically significant at 8, 10, 14, or 18 wk postvaccination. This suggests that processing RB51 into lactose based pellets and ballistically inoculating elk with these pellets does not alter the detectable elk antibody response. Also, inoculated and control animals can be accurately identified with ELISA at 4-8 weeks postvaccination. PMID- 12528443 TI - Antibodies against Mycoplasma bovigenitalium in free-living European bison (Bison bonasus) with balanoposthitis. AB - Since 1980 severe chronic balanoposthitis has been observed in free-living European bison (Bison bonasus) in the Bialowieza Primeval Forest (Poland). Sera of 50 bison with balanoposthitis and 48 clinically healthy male and 49 female bison were investigated for antibodies against Mycoplasma bovis and M. bovigenitalium by western blot analysis. Prevalence of antibodies against M. bovigenitalium was significantly higher in bison with balanoposthitis than in unaffected male bison. Mycoplasma bovigenitalium may play a role in the pathogenesis of balanoposthitis in European bison. PMID- 12528444 TI - Immobilization of swift foxes with ketamine hydrochloride-xylazine hydrochloride. AB - There is an increasing need to develop field immobilization techniques that allow researchers to handle safely swift foxes (Vulpes velox) with minimal risk of stress or injury. We immobilized captive swift foxes to determine the safety and effectiveness of ketamine hydrochloride and xylazine hydrochloride at different dosages. We attempted to determine appropriate dosages to immobilize swift foxes for an adequate field-handling period based on three anesthesia intervals (induction period, immobilization period, and recovery period) and physiologic responses (rectal temperature, respiration rate, and heart rate). Between October 1998-July 1999, we conducted four trials, evaluating three different dosage ratios of ketamine and xylazine (2.27:1.2, 5.68:1.2, and 11.4:1.2 mg/kg ketamine:mg/kg xylazine, respectively), followed by a fourth trial with a higher dosage at the median ratio (11.4 mg/kg ketamine: 2.4 mg/kg xylazine). We found little difference in induction and recovery periods among trials 1-3, but immobilization time increased with increasing dosage (P < 0.08). Both the immobilization period and recovery period increased in trial 4 compared with trials 1-3 (P < or = 0.03). There was a high variation in responses of individual foxes across trials, making it difficult to identify an appropriate dosage for field handling. Heart rate and respiration rates were depressed but all physiologic measures remained within normal parameters established for domestic canids. We recommend a dosage ratio of 10 mg/kg ketamine to 1 mg/kg xylazine to immobilize swift foxes for field handling. PMID- 12528445 TI - Molecular identification and prevalence of Dictyocaulus spp. (Trichostrongyloidea: Dictyocaulidae) in Swedish semi-domestic and free-living cervids. AB - Lungs of 102 roe deer (Capreolus capreolus), 136 moose (Alces alces), 68 fallow deer (Dama dama), and six red deer (Cervus elaphus) were examined during hunting seasons from 16 September 1997 to 1 March 2000. The aim was to determine the species composition and prevalence of Dictyocaulus lungworms in these hosts in Sweden. Worms were identified following polymerase chain reaction (PCR) amplification of the internal transcribed spacer of ribosomal DNA (ITS2), followed by hybridization with four species-specific oligonucleotides. In addition, 50 lungworms from five reindeer (Rangifer tarandus) from Norway were similarly analyzed. A total of 399 worms were recovered and analyzed representing a range of 29-128 worms per host species. All specimens from roe deer were identified as Dictyocaulus capreolus, whereas those from red deer and reindeer were identical with D. eckerti. From moose, 73 (81.1%) of the worms were identified as D. capreolus whereas 17 (18.9%) were D. eckerti. The ITS2 sequence of fallow deer lungworms differed significantly when compared with the ITS2 of D. viviparus, D. capreolus, and D. eckerti. This indicated that fallow deer in Sweden may be infected with a new genotype of Dictyocaulus spp. Consequently, a specific probe designed for the ITS2 from this Dictyocaulus sp. hybridized exclusively with samples from lungworms of fallow deer. Interestingly, no D. viviparus were found in any of these hosts. The prevalence of infection in each host was as follows: D. capreolus in roe deer (14.7%) and moose (10.6%); D. eckerti in moose (0.7%) and red deer (33.3%); and Dictyocaulus sp. in fallow deer (10.3%). Regardless of lungworm species, the overall prevalence of Dictyocaulus spp. in these hosts was 12.2%. Prevalence between male and female animals and among the different age groups did not differ significantly. Finally an enzyme linked immunosorbent assay (ELISA) specific for patent D. viviparus infection in cattle was utilized to analyze lung tissue fluids from infected animals. All samples from roe deer, red deer, and fallow deer were negative in the ELISA. However, three out of twelve (25%) samples from moose and 17 of 40 (43%) samples from cattle were positive. This indicated that moose anti-D. capreolus antibodies recognized the D. viviparus antigen and that anti-cattle immunoglobulin cross reacted with moose antibodies. PMID- 12528446 TI - Burrowing fly larvae (Philornis porteri) associated with mortality of eastern bluebirds in Florida. AB - We investigated mortality among nestling eastern bluebirds (Sialia sialis) in Polk and Highlands counties, Florida (USA) in 1999-2001. At least six species of maggots from three families of muscoid flies, Calliphoridae, Sarcophagidae, and Muscidae were found associated with the nestlings. Philornis porteri, the only species of obligate bird parasite collected, was found in the contents of two nests, in the ear canal and the musculature of the jaw of one nestling, and in the abdominal subcutis of another. This is the first record of bluebird parasitism by P. porteri. Although some nestlings were infested by tissue invading fly larvae antemortem, the role of these maggots in the overall mortality was not clear. PMID- 12528447 TI - Population dynamics of Tylodelphys destructor and Diplostomum mordax (Digenea: Diplostomidae) co-occurring in the brain of Patagonian silversides from Lake Pellegrini, Patagonia, Argentina. AB - Studies on infection patterns of diplostomid parasites in commercially exploited fishes have not been done in Patagonia (Argentina). The aim of this work was to study the population dynamics of two diplostomid species in the brain of patagonian silversides (Odontesthes hatchery), the interaction between them, and effect on health and physical condition of the hosts. Tylodelphys destructor and Diplostomum mordax metacercariae in the brain of Patagonian silversides in Lake Pellegrini were studied between January 1991 and February 1992. Tylodelphys destructor parasitized all silversides examined; prevalence of D. mordax varied between 7% and 100%. Mean intensity for T. destructor was 35-140 and for D. mordax was 3-49. Highest mean intensities of T. destructor coincided with the lowest mean intensities of D. mordax. Recruitment seems to occur from July November for T. destructor and from April-June for D. mordax, revealing a temporal segregation with inverse patterns of infection and recruitment. Tylodelphys destructor has higher intensities in the brain of the older fish, whereas D. mordax did not, suggesting another type of segregation. There were no evidences of gross pathology. No covariation between abundance of larvae and condition factor, gonadosomatic index, and gut fullness was detected. PMID- 12528448 TI - Low-dose meningeal worm (Parelaphostrongylus tenuis) infections in moose (Alces alces). AB - Parelaphostrongylosis has a rapid onset and is lethal in neonatal moose (Alces alces) when large numbers of third-stage Parelaphostrongylus tenuis larvae (L3) are given experimentally. Little is known, however, about the severity and prognosis of infections acquired naturally by accidentally ingesting terrestrial gastropods which are rarely infected and have few larvae. To investigate the relationship between infecting dose, age of moose, and severity of disease, five calves were given low doses of three to 10 L3 when five (n = 2) or 9.5 mo old (n = 3). Each of two animals initially given low doses were later challenged with a dose of 15 L3. As positive controls, two calves were given doses of 15 and 30 L3, considered to be high. All five calves given low doses showed abnormal locomotory signs at 20-28 days postinoculation (DPI) that progressively became more pronounced with hind quarter weakness and front lameness. However, after 77-130 DPI, signs diminished markedly in two of these animals and disappeared in another two. Challenge infections of 15 L3 given 199 days after initial infections had no noticeable effects although an immature worm, probably resulting from the challenge, was found in the spinal cord of one animal killed 51 days later. Two positive control animals given the high doses of 15 and 30 L3 showed moderate to severe, non-resolving, locomotory signs and had to be euthanized. Results demonstrate that single, low doses of three to 10 P. tenuis L3 cause moderate disease in moose calves but over time, some worms die and animals can recover. A degree of protection may develop against future infection. PMID- 12528449 TI - Detection of anti-parelaphostrongylus tenuis antibodies in experimentally infected and free-ranging moose (Alces alces). AB - Confirming Parelaphostrongylus tenuis infection in moose (Alces alces) and other susceptible hosts is difficult. An enzyme-linked immunosorbent assay (ELISA) was developed using the excretory-secretory (ES) products of third-stage P. tenuis larvae (ES-ELISA) and the test applied to serum samples obtained from seven moose calves (5-9.5 mo old) given infective larvae (L3) in doses approximating those likely to be received in nature (3-30 L3). Anti-P. tenuis immunoglobulin G antibodies were detected in all seven inoculated moose during the course of infection until the termination of experiment 61-243 days post-inoculation (DPI). Five animals tested between 16-25 DPI had significant antibody levels, while a sixth animal did not test positive until 46 DPI. The seventh animal was not tested until 199 DPI. Antibody levels remained elevated in all five animals that harbored adult worms at the termination of the experiment. Whereas, antibody levels showed a gradual decline in the two remaining animals, presumably because of death of worms, and antibodies were undetected in one animal at the time of necropsy. The other animal displayed an anamnestic increase in antibody level following a challenge inoculation of infective larvae. Terminal and peak optical density (OD) values detected by ES-ELISA strongly correlated with inoculation dose (r = 0.98, P = 0.02 and r = 0.95, P = 0.04, respectively) among animals harboring adult worms (n = 4) but not significantly with the number of worms recovered postmortem (peak OD, r = 0.82, P = 0.18; terminal OD, r = 0.93, P = 0.07). Unlike the ES products, use of somatic antigens of the adult worm in ELISA did not provide satisfactory results. Antibodies to P. tenuis were detectable by ES-ELISA in two of 21 free-ranging moose from an enzootic area but not from any of 23 animals from a non-enzootic area. The ES-ELISA appears to be a useful test for assessing exposure of moose to P. tenuis. PMID- 12528450 TI - Age-related hematologic changes in captive-reared houbara, white-bellied, and rufous-crested bustards. AB - Blood samples were obtained at monthly intervals between April 1994 and March 1996 from captive-bred houbara (Chlamydotis undulata macqueenii), rufous-crested (Eupodotis ruficrista gindiana), and white-bellied (Eupodotis senegalensis) bustards from 4-24 wk of age. Hematology investigations were conducted to determine age-related changes and to establish reference values for growing chicks of these species. There were significant age-related changes in hematocrit, hemoglobin, and red cell count in young birds compared with those of adults. White cell counts (lymphocytes and monocytes) were higher in juvenile birds, compared with adult values. PMID- 12528451 TI - Cerebrospinal nematodiasis in a moose in Norway. AB - A case of cerebrospinal nematodiasis in a young adult moose (Alces alces) from Telemark county, southeastern Norway, is described. The moose was found by bird hunters during January, displaying signs of severe posterior paresis. It was killed and submitted for autopsy. The carcass was emaciated, and there were skin excoriations and subcutaneous edema over both metacarpi. Histopathologic examination revealed traumatic malacia throughout the spinal cord and meningeal accumulations of mononuclear inflammatory cells and eosinophils in brain and spinal cord. Two adult female nematodes were found in sections, respectively, of the subarachnoid and subdural spaces of the thoracic spinal cord. The nematode cross sections were similar with those of the two neurotropic Elaphostrongylus species, E. rangiferi and E. cervi. The moose originated from an area overlapping the grazing area of a large population of wild reindeer (Rangifer tarandus tarandus) living on the mountain plateau of Hardangervidda, suggesting the moose was infected with E. rangiferi from reindeer. PMID- 12528452 TI - Parelaphostrongylus tenuis in captive pronghorn antelope (Antilocapra americana) in Nebraska. AB - Lesions in four captive pronghorn antelope (Antilocapra americana) naturally infected with Parelaphostrongylus tenuis in eastern Nebraska (USA) are described in this report. Animals were bright and alert with hind limb ataxia that progressed to sternal or lateral recumbency between July 28 and October 17, 1998. Animals were euthanized due to disease progression despite therapy. Multifocal decubital ulcers over bony prominences occurred in two animals and chronic unilateral otitis media was present in one animal. Histopathologic examination revealed severe Wallerian degeneration randomly scattered throughout the spinal cords of all four animals. Spinal cord sections from two animals contained adult nematode parasites consistent with P. tenuis. This is the first report of naturally occurring P. tenuis infection in pronghorn antelope. Pronghorn antelope should be considered susceptible to P. tenuis infection and contact with infected white-tailed deer as well as intermediate gastropod hosts of P. tenuis should be prevented in endemic areas. PMID- 12528453 TI - Infectious disease survey of Rio Grande wild turkeys in the Edwards Plateau of Texas. AB - State wildlife agencies have translocated thousands of wild turkeys (Meleagris gallopavo) since the 1930s to reestablish this species. Because of threats to the domestic poultry industry and wild birds, screening for selected infectious agents has become routine since the early 1980s. One of the principal sources for Rio Grande wild turkeys (M. gallopavo intermedia) for translocation purposes was the Edwards Plateau of Texas (USA). Unfortunately, turkey abundance has declined in the southern Edwards Plateau since the late 1970s. Surprisingly few studies have addressed wild turkeys in this region, perhaps reflecting its status as the heart of Rio Grande turkey range. We surveyed 70 free-living Rio Grande wild turkeys from Bandera and Kerr counties, Texas, for evidence of exposure to Salmonella typhimurium, S. pullorum, Mycoplasma gallisepticum, M. meleagridis, M. synoviae, Chlamydophila psittaci, and the avian influenza, Newcastle disease, turkey corona, and reticuloendotheliosis viruses. Of these, 80% (56) were seropositive for both M. gallisepticum and M. synoviae on the serum plate antigen test. Ten of these individuals (14% of total) were positive for M. synoviae by hemagglutination inhibition testing. All other serologic tests were negative. Two adult females sampled in Kerr County, whose body mass was significantly less than that of other adult females trapped in the area, tested positive for reticuloendotheliosis virus (REV) proviral DNA on polymerase chain reaction. Reticuloendotheliosis virus was isolated from one of these individuals. The pathogenesis, transmission, and/or population-level influences of M. gallisepticum, M. synoviae, and REV in Rio Grande wild turkeys deserves further study. PMID- 12528454 TI - Infectious disease survey of lesser prairie chickens in north Texas. AB - Lesser prairie chicken (Tympanuchus pallidicinctus) abundance, like that of most grassland birds, has declined rangewide for decades. Although habitat loss and degradation are likely ultimate causes for this decline, infectious agents, particularly microparasites, could be proximate contributors. No surveys of pathogenic bacteria or viruses have been published for this species. We surveyed 24 free-living lesser prairie chickens from Hemphill County, Texas (USA), for evidence of exposure to Salmonella typhimurium, S. pullorum, Mycoplasma gallisepticum, M. synoviae, Chlamydophila psittaci, and the avian influenza, Newcastle disease, infectious bronchitis, and reticuloendotheliosis viruses. Two of 18, and eight of 17 samples were seropositive for the Massachusetts and Arkansas serotypes of infectious bronchitis virus, respectively. Five of the eight positive individuals were juveniles, two of which were seropositive for both serotypes. All other serologic and genetic tests were negative. Because the ecological significance of these results is unknown, the pathogenesis, transmission, and/or population-level influences of infectious bronchitis and related avian coronaviruses for lesser prairie chickens deserves further study. PMID- 12528455 TI - Serologic survey for viral and bacterial infections in western populations of Canada lynx (Lynx canadensis). AB - A serologic survey for exposure to pathogens in Canada lynx (Lynx canadensis) in western North America was conducted. Samples from 215 lynx from six study areas were tested for antibodies to feline parvovirus (FPV), feline coronavirus, canine distemper virus, feline calicivirus, feline herpesvirus, Yersinia pestis, and Francisella tularensis. A subset of samples was tested for feline immunodeficiency virus; all were negative. For all other pathogens, evidence for exposure was found in at least one location. Serologic evidence for FPV was found in all six areas but was more common in southern populations. Also, more males than females showed evidence of exposure to FPV. Overall, prevalences were low and did not exceed 8% for any of the pathogens tested. This suggests that free ranging lynx rarely encounter common feline pathogens. PMID- 12528456 TI - Response of wild subantarctic fur seal (Arctocephalus tropicalis) females to ketamine and tiletamine-zolazepam anesthesia. AB - This study is the first to compare the anesthetic effects of two cyclohexamines on free-ranging subantarctic fur seal (Arctocephalus tropicalis) females. From April to July 1999, 107 females were immobilized for tooth extraction and blood sampling, using either ketamine (Ketalar, n = 58) alone or tiletamine-zolazepam (Zoletil 100, n = 49) mixture. Animals were injected intramuscularly at mean doses of 2.1 mg/kg for ketamine and 1.1 mg/kg for tiletamine-zolazepam mixture. Individual response to both drugs was highly variable. The dosage required to achieve a satisfactory level of anesthesia was smaller for subantarctic fur seals than for most other species of seals and was less for animals in better body condition. Few side effects were observed during the trials, aside from mild tremors caused by ketamine, and respiratory depression or prolonged apnea caused by tiletamine-zolazepam. We recommend use of ketamine, especially by those with little experience in anesthesia of fur seals. However, precautionary measures should be taken, such as using low doses for animals in good body condition and being prepared for anesthetic emergencies to avoid any casualties. PMID- 12528457 TI - Normal conjunctival flora in the North American opossum (Didelphis virginiana) and raccoon (Procyon lotor). AB - We documented the normal conjunctival bacterial flora from 17 opossums (Didelphis virginiana) and 10 raccoons (Procyon lotor) trapped in Manhattan, Kansas (USA) from November 1999 to January 2000. Both raccoons and opossums were free of apparent ocular disease. The inferior conjunctival sacs of each animal were swabbed for aerobic bacterial and Mycoplasma culture and polymerase chain reaction (PCR) for Mycoplasma and Chlamydia detection. All conjunctival samples were positive for one or more species of aerobic bacteria. The most common isolate from opossums was Staphylococcus spp. Other isolates included Streptococcus spp., Bacillus spp., Corynebacterium spp., and Enterococcus faecalis. The most common isolates in raccoons was Bacillus spp. Other isolates included Streptococcus spp., Staphylococcus spp., non-hemolytic Escherichia coli, and Enterococcus faecalis. Mycoplasma culture was negative in samples from opossums and raccoons. Evidence of Mycoplasma and Chlamydia presence was detected by PCR. PMID- 12528458 TI - Prevalence of antibodies against canine distemper virus among red foxes in Luxembourg. AB - Canine distemper virus (CDV) has a wide host spectrum, and during the past years, distemper has been observed in species that were previously not considered to be susceptible. In this study, we investigated the prevalence of CDV-specific antibodies in red foxes (Vulpes vulpes) sampled between May and November 1997. About 9 to 13% of the Luxembourg red fox population is positive for antibodies against CDV. Thus a sizeable proportion of red foxes has been exposed to CDV in the wild. The significance of CDV in red foxes is discussed. PMID- 12528459 TI - Atlantoaxial instability in a white-tailed deer fawn (Odocoileus virginianus). AB - On 14 March 2001, an 8 mo old, male white-tailed deer (Odocoileus virginianus) was found in lateral recumbency exhibiting neurologic signs including inability to rise, opisthotonus, paddling, and respiratory distress. There was evidence of minor cranial trauma. Postmortem examination revealed atlantoaxial instability with ventral deviation of the axis due to malformation of the caudal atlas and cranial axis. Given the age of the fawn, the instability was assumed to be congenital with minor trauma inducing severe, acute neurologic signs. PMID- 12528460 TI - [A non-statistical approach to the prediction of regular structures in proteins by the example of alpha-helices]. AB - A new approach to the analysis of regular structures in proteins that is based on the method of molecular mechanics is proposed. The method uses only the information about the amino acid sequence. The alpha-helical conformation was simulated using the ICM program of molecular mechanics. Energy profiles of the sequences in the alpha-helical conformation, spanning the entire polypeptide chain, were plotted for eight proteins from the Protein Data Bank. The regions of each profile that exhibit energy minima were found to correspond to the alpha helical regions of the real spatial structure of the protein. Twenty-four out of 25 helices were distinctly pronounced, which indicates a rather high accuracy of the prediction. The energy profiles also help reveal the short regions that correspond to 3/10-helices and the turns that include local alpha-helical conformations. Unlike the known statistical methods of prediction, this method makes it possible to establish the physical principles of the formation of alpha helical conformations. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2002, vol. 28, no. 6; see also http://www.maik.ru. PMID- 12528461 TI - [Stereochemistry of the reductive amination of 4-oxoproline derivatives with glycine esters]. AB - An amination of 4-oxoproline derivatives with glycine methyl or benzyl ester and sodium cyanoborohydride led to the mixtures of corresponding diastereomeric 4-cis and 4-trans-glycinoproline derivatives. We found that the ratio of diastereomers mainly depends on the structure of 4-oxoproline ester groups and, to a lesser extent, on the structure of N-acyl substituents. The best results were achieved with tert-butyl ester group; it ensured good yields of the amination products and the greatest prevalence of 4-cis-isomers. The structure of ester group in glycine molecule only scarcely affected the resulting ratio of N-(N benzyloxycarbonylglycyl)-4-glycinoprolines. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2002, vol. 28, no. 6; see also http://www.maik.ru. PMID- 12528462 TI - [The effect of pyrophosphate analogues on the inorganic pyrophosphatase from Escherichia coli]. AB - The effect of inorganic pyrophosphate analogues on the enzymic activity of inorganic pyrophosphatase from E. coli was studied. Hypophosphoric and diphosphonic acids were shown to inhibit inorganic pyrophosphatase, whereas pyrophosphorous acid exerts almost no effect on the hydrolysis of inorganic pyrophosphate. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2002, vol. 28, no. 6; see also http://www.maik.ru. PMID- 12528463 TI - [Ligands with specific affinity to a specific sequence of DNA base pairs. XI. The synthesis and binding to DNA of bis-netropsins with the C-ends of their netropsin fragments tethered by tetra- or pentamethylene linkers]. AB - Bis-Netropsins with the C-ends of their netropsin fragments tethered via tetra- or pentamethylene linkers and with Gly or L-Lys-Gly residues on their N-ends were synthesized. The footprinting technique was used to study the specificity of bis netropsin binding to the specially constructed DNA fragments containing various clusters of A.T pairs. It was found that the linker length affects the binding of bis-netropsins, with the tetramethylene linker providing better protection than the pentamethylene linker. It was shown that the newly synthesized bis-netropsins bind tighter to the 5'-A4T(4)-3' sequence, whereas the bis-netropsin with a linker between the netropsin N-ends binds better to 5'-T4A(4)-3' sequences. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2002, vol. 28, no. 6; see also http://www.maik.ru. PMID- 12528464 TI - [Neoglycoconjugates based on dendrimeric poly(aminoamides)]. AB - Neoglycoconjugates containing 4, 8, 16, 32, and 64 terminal residues of B disaccharide (BDI) or N-acetylneuraminic acid (Neu5Ac) attached to poly(aminoamide)-type dendrimers (PAMAMs) were synthesized. The ability of BDI conjugates to bind natural xenoantibodies (anti-BDI antibodies) and the ability of Neu5Ac conjugates to inhibit the hemagglutinin-mediated adhesion of influenza virus were studied. The biological activity of PAMAM conjugates turned out to be higher than that of free carbohydrate ligands, but less than that of multivalent glycoconjugates based on other types of synthetic polymeric carriers. A conformational analysis of PAMAM matrices and resulting conjugates was performed to determine the statistical distances between carbohydrate ligands. The computations revealed the tendency of the PAMAM chains toward compaction and formation of dense globules. The process results in a decrease in the distances between the carbohydrate ligands in the conjugates and, hence, could affect the ability of glycoconjugates to efficiently bind the polyvalent carbohydrate recognizing proteins. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2002, vol. 28, no. 6; see also http://www.maik.ru. PMID- 12528465 TI - [A convenient synthesis of retinal derivatives with a modified trimethylcyclohexene ring]. AB - A method of simultaneous one-stage synthesis of three retinal derivatives (5,6 dioxo-5,6-seco-, 5,6-dihydro-5,6-epoxy-, and 4-oxoretinal) was proposed, with the yield of the first derivative being approximately 50%. These compounds are useful tools for studying the antitumor activity of retinoids, the reconstituted bacteriorhodopsin analogues with changed parameters of photocycle, and the reactivity of retinal derivatives in the processes of oxidation by molecular oxygen. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2002, vol. 28, no. 6; see also http://www.maik.ru. PMID- 12528467 TI - [The determination of total concentration and activity of antioxidants in foodstuffs]. AB - We developed a new method for the analysis of active antioxidants that is based on their reactions with the ABTS+. cation radical obtained by oxidation of ABTS, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt. The feasibility of this method was confirmed by electrochemical and kinetic studies of model antioxidants. ABTS+. was shown to react rapidly with active and slowly with weak antioxidants, which allows it to be used as a model radical for the quantitative determination of the total content of natural antioxidants (antioxidant equivalent) in natural extracts and wines. Another analytical method based on the competitive oxidation of Pyrogallol Red (a detecting molecule) and the examined antioxidants by radicals derived from peroxynitrite was used for measuring the relative activity of antioxidants. A combination of both methods helped measure the total concentration of antioxidants and their average specific activities (per molecule of active compound) in extracts from grape, olive, and tomato and concentrates of various popular beverages (wines, beers, and juices), as well as in the commercial concentrated food product Kreto-A, made from grape, red wine, tomato, and olive. Red wine and red grape juice were shown to be the most rich in antioxidants (up to 20 mM), with their activity being similar to that of polyphenols. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2002, vol. 28, no. 6; see also http://www.maik.ru. PMID- 12528468 TI - [Prostaglandin E2 synthases]. AB - Prostaglandin E2 (PGE2) is widely distributed in various tissues, and exhibits various biologically important activities. PGE2 synthase (PGES) catalyzes conversion of COX-derived PGH2 to PGE2. It now appears that there are at least three distinct types of PGES in mammals. We identified two distinct glutathione dependent PGESs. Cytosolic PGES (cPGES), known as p23, is constitutively and ubiquitously expressed and predominantly converts COX-1-derived PGH2 to PGE2. We find that the regulation of cPGES/p23 activity in cells depends on its association with hsp90. Microsomal PGES-1 (mPGES-1), identical to MGST1-L1, is an inducible perinuclear enzyme that is functionally linked with COX-2 in marked preference to COX-1. COX-2 and mPGES-1 are essential components for delayed PGE2 synthesis, which may be linked to inflammation, fever, osteogenesis, and even cancer. Most recently, glutathione-nonspecific mPGES-2, homologous to glutaredoxin and thioredoxin, was identified. These PGESs seem to be a potential novel target for drug development. PMID- 12528466 TI - [Synthesis and pharmacological activity of betulin dinicotinate]. AB - The assignment of NMR resonances of lupane triterpenoids was refined by the example of 3,28-dinicotinoylbetulin, obtained by acylation of betulin. Hepatoprotective, untiulcer, antiinflammatory, reparative, and anti-HIV activities were found for the compound. In addition, it was demonstrated to have immunomodulatory activity, for the first time detected among lupane triterpenoids. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2002, vol. 28, no. 6; see also http://www.maik.ru. PMID- 12528469 TI - [Pharmacotherapy of osteoporosis]. AB - Osteoporosis with increased risk of bone fracture is a disabling syndrome that naturally occurs as long as one ages and moves on two legs. Recent progress in bone cell biology has shed light on the mechanisms underlying the anti osteoporotic properties of drugs that have been in use for a long time, providing a fresh stage for novel pharmacotherapies. In addition, large scale clinical trials developed in the past decade appear not only to rationalize the clinical utilities of these drugs but also to provide new concepts for the development of new therapeutic modalities. Progress in the fields of basic and clinical research field is briefly reviewed herein. PMID- 12528470 TI - [Meloxicam (Mobic): a review of its pharmacological and clinical profile]. AB - Meloxicam (Mobic) is a new nonsteroidal anti-inflammatory drug (NSAID) derived from enolic acid, exhibiting selectivity for cyclooxygenase (COX)-2 over COX-1. Meloxicam has shown potent anti-inflammatory and analgesic activity together with low gastrointestinal toxicity in animal models. It is a potent inhibitor not only of acute exudation in adjuvant arthritis in the rat, but also of bone and cartilage destruction. The therapeutic range of meloxicam in the rat, with regard to inhibition of adjuvant arthritis, was several times greater than that of other NSAIDs. Meloxicam in therapeutic doses was found to have no effect on bleeding time or platelet aggregation in healthy volunteers. In clinical studies, meloxicam has shown reliable efficacy against rheumatoid arthritis, osteoarthritis, lumbago (low back pain), scapulohumeral periarthritis, and neck shoulder-arm syndrome with low gastrointestinal toxicity. PMID- 12528471 TI - [Novel mode of action of ribavirin (Rebetol), a drug for the treatment of chronic hepatitis C: inducting the mutation of RNA viruses]. AB - Ribavirin (Rebetol) is an antiviral agent used in combination with interferon alpha-2b (IFN alpha-2b) for the treatment of chronic hepatitis C. Ribavirin has been reported to have a broad-spectrum antiviral activity mainly against RNA viruses. The effect of ribavirin was potentiated when administered in combination with IFN alpha-2b in an antiviral assay using bovine viral diarrhea virus as a surrogate for hepatitis C virus (HCV). Inhibition of host inosine monophosphate dehydrogenase and inhibition of RNA-dependent RNA polymerase (RdRp) of RNA viruses have been reported as the modes of action of ribavirin. Recently, ribavirin has been shown to induce mutation as template for newly generated RNA after uptake in RNA by RdRp of poliovirus, which is an RNA virus as in the case of HCV. It has also been shown that the infectivity of viruses is drastically reduced by the very slight increase in mutations induced by ribavirin. This effect as a mutagen on RNA viruses is a novel mode of ribavirin, and it is thought necessary to classify ribavirin into a new antiviral drug class. PMID- 12528472 TI - [Pharmacological and clinical properties of alendronate sodium hydrate]. AB - Alendronate (alendronate sodium hydrate; Bonalon Tablet, 5 mg) is a nitrogen containing bisphosphonate, which combines with the bone surface and reduces osteoclast-mediated bone resorption. It is a third-generation bisphosphonate compound, specifically distributed on the surface of bone resorption and taken into osteoclasts. Under the closed circumstances which is formed with osteoclast and the bone surface, alendronate becomes detached from the bone surface and taken into osteoclast since acid released from osteoclast leads to pH decrease (acidified). The uptaken alendronate blocks the pathway of mevalonic acid synthesis, which is cholesteric synthesis, inhibits the prenylation of GTP binding protein, and decreases the osteoclast's function by influencing the cytoskeleton. This restraint of alendronate in bone resorption against osteoclasts is reversible, showing no cytotoxicity at more than hundredfold concentration level at which action occurs. Alendronate is an agent for the treatment of osteoporosis that has established safety with regards to bone quality since it neither inhibits bone calcification nor influences fracture healing in chronic administration. The most serious morbidity in osteoporosis is developing fractures. The efficacy of alendronate on restraining fracture, as well as on increase in BMD, is evidenced in Japan. Recently, in addition to senile or postmenopausal osteoporosis, drug-induced osteoporosis, such as steroid induced osteoporosis, has attracted attention. In this regard, alendronate has been found to be an effective agent for the treatment of osteoporosis overseas, being approved in over 90 countries and used by more than 4.5 million patients. This review will give an outline of alendronate, the preparation to have introduced a concept of Evidence Based Medicine earlier, from pharmacodynamic action to clinical efficacy. PMID- 12528473 TI - [Interferon alfacon-1 (Advaferon): a novel synthetic interferon for the treatment of hepatitis C, its pharmacological and clinical profile]. AB - Interferon alfacon-1 (Advaferon, also referred to as consensus interferon) is a novel synthetic recombinant type-I interferon (IFN) developed by comparing the amino acid sequences of several natural IFN-alpha subtypes and assigning the most frequently observed amino acid in each corresponding position to generate a consensus molecule. Interferon alfacon-1 binds with high affinity to type-I IFN receptors and has greater biological activity than naturally occurring IFN-alpha as assessed by its increased antiviral, anti-proliferative, and natural killer cell activities, as well as its stronger IFN-stimulated gene induction. In a multicenter, randomized, controlled study, the safety and efficacy of interferon alfacon-1 in comparison with lymphoblastoid interferon-alpha [IFN-alpha (NAMALWA)] was evaluated in patients infected with high-titer chronic hepatitis C virus (HCV). Interferon alfacon-1 (18 MIU) was superior in efficacy without additional toxicity to IFN-alpha (NAMALWA) (9 MIU) in high-titer chronic HCV patients, particularly those infected with genotype 1b. A multicenter open-label study showed that treatment with interferon alfacon-1 (12 MIU) was an effective and tolerable therapy in chronic HCV patients with low viral titers. Another multicenter open-label study showed that re-treatment with interferon alfacon-1 (18 MIU) was an effective and tolerable therapy in chronic HCV patients who relapsed after traditional IFN therapy. Collectively, these clinical studies indicate that interferon alfacon-1, as a novel synthetic interferon, may be a useful therapeutic alternative for the effective treatment of hepatitis C. PMID- 12528474 TI - [Pharmacological action and clinical effects of falecalcitriol, a highly potent derivative of active vitamin D3]. AB - Much effort has been made to create highly potentiated active vitamin D for better clinical applications and falecalcitriol was successfully synthesized as one of such candidates with highly potent and long-lasting effects. Its chemical structure has a calcitriol side chain modification in which both methyls at positions C-26 and C-27 are substituted by tri-fluoromethyls. The mechanism for its strong and long-lasting effects is probably due to altered side chain metabolism and decreased inactivation. Although C-24 position hydroxylation catalyzed by Cyp24 inactivates calcitriol, falecarcitriol is metabolized to C-23S hydroxylated metabolite by the same enzyme Cyp24 and this metabolite still has strong activity. Stronger action of falecalcitriol has been shown in target organs or cells of active vitamin D such as bone, parathyroid cells, and keratinocytes, when compared with calcitriol, the endogenous active form of vitamin D. Daily oral administration of falecalcitriol at doses lower than those required for calcitriol has been shown to have clinical effects for the treatment of diseases such as hyperparathyroidism due to chronic renal failure (2 degrees HPT), rickets, osteomalacia and hypoparathyroidism. The comparative study with alfacalcidol showed its specific action on parathyroid hormone suppression and better improvement of bone metabolism markers in 2 degrees HPT patients. PMID- 12528475 TI - From skeptic to convert: a physical therapist's story. Improved quality of life is the only acceptable outcome for patients. PMID- 12528476 TI - Developing a telehealth business plan. Anticipating challenges and devising ways to meet them leads to successful services. PMID- 12528477 TI - Telepharmacy offers hope for rural hospitals. National pharmacist shortage prompts innovative approach to hospital-based services in Washington state. PMID- 12528478 TI - Pioneering corrections service offers tips for success. UTMB's success tied to close alignment between information technology and clinical services groups. PMID- 12528479 TI - Interactive home telehealth: moving from cost savings to reimbursement. Creative, proactive strategies help agencies turn telehealth into a revenue generator. PMID- 12528480 TI - Risk management for telemedicine providers: how to stay out of jail. Six standards define whether arrangements fall within safe harbors. PMID- 12528481 TI - India telemedicine venture seeks to improve care, increase access. Spurred by government initiatives, private industry invests in linked hospital network. PMID- 12528482 TI - Iris scanning: the eye of the storm. Several industries have hastened development of authentication technologies-including healthcare. PMID- 12528483 TI - CA Blue Cross using quality to reward physician performance. PMID- 12528484 TI - Use risk adjustment to make sense of clinical trends. PMID- 12528485 TI - Integrated software system makes coding easier for Missouri hospital. PMID- 12528486 TI - The good, the bad, the ugly--Texas ranks hospital care. PMID- 12528487 TI - Treating varicose veins with transilluminated powered phlebectomy. AB - In the past, patients endured hours of surgery, many incisions, and multiple scars to manually remove painful, unsightly, swollen varicose veins. Patients now have a new treatment option. Endoscopic resection and ablation of superficial varicosities is possible using a powered vein resector, irrigated illuminator, and tumescent anesthesia. This article describes this new technique, called transilluminated powered phlebectomy, and explains how the procedure reduces the number of incisions and provides direct visualization of the veins as they are resected. PMID- 12528488 TI - Anterior cervical discectomy and fusion for cervical disc disease. AB - Anterior cervical discectomy and fusion (ACDF) is a surgical procedure proven successful in the treatment of symptoms caused by cervical degenerative disc disease that is unresponsive to conservative therapy. Retrospective studies of patients who have undergone ACDF indicate that this procedure has a high rate of success for relieving symptoms and a low rate of associated complications. This article discusses normal cervical spine anatomy, the pathology of degenerative cervical disc disease, and perioperative nursing care for patients undergoing ACDF. Cortical ring allograft bone grafting and internal fixation with cervical locking plates also is discussed. PMID- 12528489 TI - Complementary and alternative medicine use by surgical patients. AB - This national multisite study examined complementary and alternative medicine (CAM) use by surgical patients during the two weeks preceding their surgery. A simple descriptive design consisting of anonymous self-report questionnaires was used to measure CAM use and determine participant demographics. A purposive sample of adult, English-speaking, elective outpatients from 16 hospitals representing seven US regions was used. Data were collected randomly at each site. Sixty-seven percent of participants disclosed use of all types of CAM; 27% consumed herbs, 39% used dietary supplements, 54% took vitamins, and 1% reported use of homeopathics. Thirty-four percent of participants reported using CAMs that potentially can interact with anesthetics and inhibit coagulation. The results of this study suggest that perioperative nurses should encourage patients to disclose CAM use and should educate patients about anesthetic interactions that may contribute to bleeding. PMID- 12528490 TI - Nursing school curricula and hospital-based training programs. AB - Perioperative nursing is at the threshold of an unprecedented shortage that may affect the profession for years to come as the existing workforce ages and nursing schools continue to marginalize perioperative curricula. The purpose of this study is to investigate perioperative staffing issues, preceptorship training programs, and nursing school curricula in the metropolitan area surrounding the city of San Jose, Calif. Results show that 77% of the ORs surveyed had vacant positions and experienced moderate to severe difficulty in filling these vacancies. Results for the schools of nursing show that 83% do not offer preceptorships or clinical internships in the perioperative setting. Recommendations regarding the development of professional, community, and academic partnerships to help address the perioperative nursing shortage are presented. PMID- 12528491 TI - A precepted perioperative elective for baccalaureate nursing students. AB - A major factor contributing to the recruitment crisis in perioperative nursing is the lack of perioperative experiences offered to nursing students. A clinical elective designed to provide interested baccalaureate nursing students with OR experience was implemented at one university. A faculty member collaborated with perioperative nurses from area hospitals to provide a 36-hour, precepted perioperative experience. The course allows students to learn about the perioperative nursing role and helps them develop nursing skills. Students acknowledged that this brief experience was adequate to meet their learning needs and help them decide whether they wanted to pursue further a career in perioperative nursing. PMID- 12528492 TI - Increased efficiency through OR redesign and process simplification. AB - Short turnover times and efficient use of time and personnel are hallmarks of an efficiently run OR. When delays occur, time can be lost and the workday prolonged considerably. This article describes changes made in neurosurgical services at Utah Valley Regional Medical Center, Provo. These changes involved redesign of the neurosurgical suite and simplification of processes, which led to a 33% to 55% decrease in turnover times and an increase in team efficiency and the number of nurses proficient in the specialty. PMID- 12528493 TI - Ethics in perioperative practice--values, integrity, and social policy. AB - Though often difficult, ethical decision making is necessary when caring for surgical patients. Perioperative nurses have to recognize ethical dilemmas and should be prepared to take action based on the ethical code outlined in the American Nurses Association's (ANA's) Code of Ethics for Nurses with Interpretive Statements. In this final article of a nine-part series that is designed to help perioperative nurses relate the ANA code to their own area of practice, the author looks at the ninth provision, which emphasizes the responsibility of professional nursing associations to maintain the value and integrity of the profession. PMID- 12528494 TI - Recommended practices for environmental cleaning in the surgical practice setting. PMID- 12528495 TI - Healing patients on a spiritual, as well as physical level. PMID- 12528496 TI - How to use medical devices safely. AB - Perioperative nurses play an important role in the prevention of medical device problems. Be proactive and prevent the harm device problems can cause. Nurses can have a profound effect on eliminating this type of patient harm in the perioperative setting, and through reporting, they can prevent reoccurrence and harm to other patients. PMID- 12528498 TI - Consultants: managing them means measuring them. PMID- 12528499 TI - Hospital pantomime. Break a leg. PMID- 12528500 TI - NHS Christmas. It's a family affair. AB - How strong an influence is childhood on subsequent choice of career? What is the power of values absorbed at an early age? Joanna Lyall spoke to individuals who have achieved eminence in the field about their early influences and memories of Christmases past. PMID- 12528502 TI - [Studies on investigation and utilization of medicinal plant resources for sustainable development in Guizhou, China]. AB - OBJECTIVE: To gain a clear idea of the resources of medicinal plants and their pharmaceutical identification in Guizhou, China, and to conserve, develop, and uilize them effectively. METHOD: Investigation on the spot with a statistical analysis. The technical route is "Investigation and identification-->Conservation and development-->Method, technology and ways for protection, propagation and utilization-->GAP cultivated pilot and assessment of medicinal materials and their products-->Comprehensive evaluation and results". RESULT: According to the incomplete statistics, there are 3924 varieties of medicinal materials from plants, accounting for 91.47% of the total in Guizhou. Among them there are 275 families, 1384 genera and 2987 species of vascular bundle plants, accounting for 43.1% of all species. There are 200 species of pteridophyta and 2802 spermatophyta. CONCLUSION: Investigation of wild medicine species has been proceeded, as well as introduction and research of their propagation techniques and production. Guizhou is rich in medicinal plant resources. However, their species and population are reducing progressively. Effective measures must be taken to protect the germ plasma resources and put them into rational uses for the utilization and sustainable development. Rare and endemic medicine species must be introduced for conservation, propagation and realization of GAP pilot cultivation. Resources must be used in an economical way and products must be made with a high technological content. PMID- 12528501 TI - [Pharmacological study on Tianxiong (tuber of Aconitum carmichaeli Debx.), a Chinese drug for reinforcing the kidney yang retail in Hong Kong market]. AB - OBJECTIVE: To explore the pharmacodynamic mechanism of Tianxiong (Aconitum carmichaeli) in tonifying the kidney and supporting Yang, so as to provide evidences for further development of new drugs treating Yang-eficiency of the kidney. METHOD: Observing parameters such as visceral index, survival time of low temperature swimming for hydrocortisone-induced Yang-deficiency model mouse and testis-removed kidney-deficiency model rat. RESULT: The decoction of processed Tianxiong could strengthen the antifatigual ability and prolong the survival time of low-temperature swimming for mice, and promote immunization in rats. CONCLUSION: Tianxiong is able to reinforce the kidney Yang, which reconforms the conclusion of "replenishing the fire of vital gate and the Qi of kidney" recorded in Chinese historical literature and proved by overseas clinical practice. PMID- 12528503 TI - [Studies on tissue culture of Dendrobium chrysotoxum Lindl in vitro]. AB - OBJECTIVE: To set up a system for the culture of Dendrobium chrysotoxum in vitro. METHOD: Tissue culture, fire fly luminescence and phenol-H2SO4 method. RESULT: The embryo could germinate with or without light, the MS, 1/2MS, B5, N6 mediums are suitable to the growth and the differentiation of sprout with light, 0.5 mg.L 1 NAA and 1 mg.L-1 6-BA, and ATP have regular changes, the content of polysaccharide was 2.833% in plant and 7.254% in sprout. CONCLUSION: The light has no effects on the embryo germination, but the phytohormone, nitrogen source and organized elements are important to the growth and differentiation of the sprout which should be transferred to the MS, 1/2MS, B5, N6 mediums in time supplemented with NAA [symbol: see text] 6-BA, ATP may be served as the dynamic indication of nourishment demand in the plant. The content of polysaccharide in the sprout is higher and can be utilized. PMID- 12528504 TI - [Studies on polysaccharides in different development stages of Armillaria mellea (Vahl.: Fr.) Quel]. AB - OBJECTIVE: To study the polysaccharides in different development stages of Armillaria mellea. METHOD: Polysaccharides in rhizomorph, fruit-body, mycelia and its fermenting liquor of A. mellea were extracted, isolated and purified. The properties contents, molar ratio and molecular weight of the polysaccharides were determined by IR spectra, HPLC, GPC and gel chromatography. RESULT: The polysaccharides in mycelia and its fermenting liquor contained only glucose, the sugar contents were 9.00% in mycelia and 0.87 g.(100 ml)-1 in fermenting liquor respectively. The polysaccharides in both rhizomorph and fruit-body consisted of glucose and xylose, and the molar ratio was 1:14 in rhizomorph and 1:10 in fruit body respectively; the polysaccharide contents were 1.12% in rhizomorph and 2.27% in fruit-body. The molecular weight of these polysaccharides was about 10,000 70,000. CONCLUSION: The experimental results supply important scientific data for developing A. mellea as a medicine. PMID- 12528505 TI - [Researches on morphology and anatomy of root system of Dendrobium nobile Lindl]. AB - OBJECTIVE: To understand the morphological and anatomical characteristics of Dendrobium nobile so as to provide scientific basis for its domestication and cultivation. METHOD: The root's morphological development and interior structure were investigated. RESULT AND CONCLUSION: Dendrobium has peculiar structure which determines that this plant requires a growing environment with high water and air ventilation properties. PMID- 12528506 TI - [Effects of processing on antioxidation of Radix et Rhizoma Rhei and Rhizoma Polygoni Cuspidati]. AB - OBJECTIVE: To investigate the effects of Processing on antioxidation of Radix et Rhizoma Rhei and Rhizoma Polygoni Cuspidati. METHOD: The oxygen free radicals generation system and mouse liver homogenate lipid peroxidation in vitro were used. RESULT: The processed products of Radix et Rhizoma Rhei and Rhizoma Polygoni Cuspidati could scavenge superoxide radical (O2-.) generated through hypoxanthine-oxidase system and (.OH) generated through Fenton action, and inhibit lipid peroxidation induced by hydroxyl radical generation system, respectively. There existed significant differences among the different processed products. CONCLUSION: After Radix et Rhizoma Rhei and Rhizoma Polygoni Cuspidati were processed, their effects became weaker. PMID- 12528507 TI - [Studies on chemical constituents of Maackia amurensis Rupr. et. Maxim]. AB - OBJECTIVE: To study the chemical constituents of EtOH extract from heartwood of Maackia amurensis. METHOD: Silica gel column chromatography and Sephadex LH-20 were employed to separate, and the compounds obtainedere elucidated by means of UV, IR, MS, 1H NMR, 13C NMR. RESULT: 4', 8-dihydroxyl-7-methoxylisoflavone (I), tectorigenin(II), glycitein(III) and resveratrol (IV) were isolated and identified. CONCLUSION: Among them, Compound I was a new isoflavone and compound III was obtained from M. amurensis for the first time. PMID- 12528508 TI - [Studies on chemical constituents from Chinese medicinal plant Hemistepta lyrata Bunge]. AB - OBJECTIVE: To separate and identify the chemical constituents of the whole plant. METHOD: The compounds were extracted with solvents, isolated by column chromatography and identified by spectroscopic methods, such as IR, MS and 1H NMR. RESULT: Five compounds were identified as hentriantane I, taraxasteryl acetate II, taraxasterol III, beta-sitosterol IV, and stigmasterol V. CONCLUSION: The compounds III and V were obtained from the plant for the first time. PMID- 12528509 TI - [Isolation and elucidation of chemical constituents of Shandanshaoyao Decoction(I)]. AB - OBJECTIVE: To study the chemical constituents of Shandanshaoyao Decoction. METHOD: The sample was prepared using boiling water, then precipitated by 75% ethanol. The ethanol soluble fraction was extracted by chloroform, ethyl acetate and n-butanol respectively. The extracts were isolated using silica gel and polyamide column chromatography, and the chemical constituents were identified by chemical and spectroscopic methods. RESULT: Six compounds were isolated from the chloroform and ethyl acetate extracts, and elucidated as tanshinone IIA, benzoic acid, ferulic acid, cryptotanshinone, beta-stitosterol and ursolic acid respectively. CONCLUSION: A preliminary study on the chemical constituents in traditional Chinese compound recipe Shandanshaoyao Decoction is reported. PMID- 12528510 TI - [Studies on polysaccharide of Cynomorium songaricum Rupr]. AB - OBJECTIVE: To study the acidic heteropolysac charides of Cynomorium songaricum. METHOD: The polysaccharides were purified by Sephadex G-100 and G-150 gel column chromatography. Purity and molecular weight of the polysaccharides were determined by high performance gel permeation chromatography; neutral sugars composition were identified by PC, TLC and GC. Uronic acids were determined by carbazole method. RESULT: The molecular weight of SYP-A and SYP-B were estimated to be 3.1 x 10(5) and 2.8 x 10(5) respectively, and neutral sugars were composed of galactose, glucose, arabinose, rhamnose, mannose and ribose. The molar ratio for SYP-A and SYP-B were 5.1:4.1:1.6:1.0:0.5:0.3 and 5.2:4.2:1.5:1.0:0.5:0.2 respectively, The contents of uronic acids were 10.7% and 10.5% respectively. CONCLUSION: SYP-A and SYP-B are homogeneous acidic heteropolysaccharides. PMID- 12528511 TI - [Contents of total flavonoids in Rhizoma Arisaematis]. AB - OBJECTIVE: Comparing the contents of total flavonoides of Rhizoma Arisaematis, which collected in different time, regions, different varieties and processed. METHOD: Determining the contents by ultraviolet spectro-photometry. RESULT AND CONCLUSION: The contents were found in the following sequence: 1. the end of July, the begin of July, August, September; 2. Beijing, Shanxi, Sichuan, Anhui; 3. Arisaema erubenscens, A. heterophyllum, A. amurense; 4. unprocessed product, processed product. PMID- 12528512 TI - [Protective effect of tanshinone on injured cultured PC12 cells in vitro]. AB - OBJECTIVE: To investigate the protective effects of tanshinone on ischemia-like injury models. METHOD: Six ischemia models including hypoxia, hypoglucose, oxidant injury, calcium overload, nitric oxide neurotoxicity and glutamic acid injury were used to assay the anti-ischemic roles of tanshinone in cultured PC12 cells, by using morphological examination, MTT staining and LDH measurement. RESULT: It was found that 3. 125-200 microliters.ml-1 tanshinone possessed obvious protective effects on PC12 cells from six ischemia-like injury models. Tanshinone could increase the number of life cells and decrease LDH activity significantly, particularly in hypoxia and caffeine injured model. CONCLUSION: Tanshinone protected PC12 cells from all injury models effectively in vitro. Its action may mainly focus on inhibiting development of the primary period of ischemia injury and calcium overloading injury. PMID- 12528513 TI - [Protective effect of Panax quinquefolium 20s-proto-panaxdiolsaponins on acute myocardial infarction in dogs]. AB - OBJECTIVE: To study the protective effects of Panax quinquefolium 20s protopanaxdiolsaponins extracted from leaves of P. quinquefolium (PQDS) on acute myocardial infarction(AMI) in dogs. METHOD: The parameters of myocardial infart size, the serum CK and LDH activity, myocardial metabolism, free radicals and coronary circulation etc were determined by using the model of ligation of LAD in the anaesthetized open-chest dogs. RESULT: In dogs treated with PQDS(in a dosage of 10 and 20 mg.kg-1 i.v. infusion), the myocardial infarct size, the activity of serum CK, LDH and the contents of serum FFA and LPO were decreased, whereas the activity of serum SOD and GSH-Px increased markedly. At the same time, myocardial blood flow was increased and coronary vascular resistance decreased significantly. CONCLUSION: PQDS has protective effect on myocardial ischemia by modifying metabolic dysfunction of FFA, inhibiting oxygen free radical mediated peroxidation of membrane lipids, enhancing endogenous antioxidase activity and increasing myocardial blood supply. PMID- 12528514 TI - [Comprehensive evaluation of cuanhuning injection]. PMID- 12528515 TI - [Advances in study on pharmacological effects of Epimedium]. AB - OBJECTIVE: To review the progress in the research of the pharmacological activities of active ingredients and extracts of Epimedium. METHOD: Effects of Epimedium on cardiovascular system, circulatory system, immune system, genital system and bone marrow system, etc were reviewed, based on the recent 10 years pharmacological experimental studies. CONCLUSION: The results provided a rational foundation for the further development and utilization of Epimedium. PMID- 12528516 TI - [Comparative study on three types of chemicals extracted from tubers of populations of Pinellia ternata (Thunb.) Breit]. AB - OBJECTIVE: To study the genetic diversity of Pinellia ternata. on the chemical composition. METHOD: Fifteen populations of Pinellia ternata. originated mostly from middle and lower reaches of Yangtze River valley were collected and cultivated in Weigang Experimental Farm of Nanjing Agricultural University with same cultural conditions, and contents in three types of index chemicals extracted from tubers of the populations have been compared continuously. RESULT AND CONCLUSION: The difference in contents of beta-sitosterol and Val and Arg extracted from tubers of the populations has been observed, and those contents are also different with harvesting time. But the mean contents of each of the three types of chemicals in autumn are all higher than those in summer respectively. PMID- 12528517 TI - [Studies on investigation and utilization of medicinal plant resources for sustainable development in Guizhou, China (to be continued)]. PMID- 12528518 TI - [Determination of jujubaside A in semen ziziphi spinosae by RP-HPLC]. AB - OBJECTIVE: To develop a new method for the determination of Jujubaside A in Semen Ziziphi Spinosae. METHOD: A RP-HPLC method was set up, using Hypersil-18 column, acetonitrile: water(30:70) as mobile phase with a flow rate of 0.8 ml.min-1. RESULT: The calibration curve was linear in the range of 0.076-0.456 mg. The average recovery was 93.5%. CONCLUSION: The method is appropriate for the determination of Jujubaside A. PMID- 12528519 TI - [A study on lyophilization injection of cucurbitacin polylactic acid nanoparticles]. AB - OBJECTIVE: To prepare stable and redispersable Lyophilization injection of cucurbitacin polylactic acid nanoparticles(Cu-PLA-NP). METHOD: An optimal supporting agent was chosen to prepare the Cu-PLA-NP lyophilization injection. The physical and chemical properties of the injection were evaluated. RESULTS: The shape, diameters, association ratio, drug loading and pH of Cu-PLA-NP colloidal solution and lyophilization injection were not changed. The content of water consisted with the requirement of lyophilization injection. The critical relative humidity was 69.52%. The stability was good. CONCLUSION: It is practicable to prepare Cu-PLA-NP lyophilization injection with proper formulas and preparation processes. PMID- 12528520 TI - [Studies on antiviral flavonoids in yinqiaosan powder]. AB - OBJECTIVE: To study the chemical constituents in Yinqiaosan Powder that have antiviral action on influenza. METHOD: Isolation of the constituents by different kinds of column chromatography and elucidation of their structures by chemical and spectral methods. RESULTS: Seven flavonoids were obtained, and six of them were identified as linarin, acacetin, hesperidin, isoquiritigenin, isoquiritin and hyperoside respectively. CONCLUSION: Linarin and acacetin were isolated from Yinqiaosan Powder for the first time. PMID- 12528521 TI - [Studies on chemical constituents in fruit of Lycium barbarum L]. AB - OBJECTIVE: To study the chemical constituents in the fruit of Lycium barbarum. METHOD: The chemical constituents were isolated by column chromatography and identified by spectral data. RESULT: The compounds obtained were identified as scopoletin(I), beta-sitosterol(II), p-coumaric acid(III), glucose(IV), daucosterol(V) and betaine(VI). CONCLUSION: Compounds III, IV and V were isolated from Lycium barbarum for the first time. PMID- 12528522 TI - [Secondary metabolites of Gliocladium sp., a growth accelerating fungus for Anoectochilus roxburghii (Wall.) Lindl]. AB - OBJECTIVE: To study the secondary metabolites of fungus Gliocladium sp. that helps accelerate the growth of A. roxburghii. METHOD: Compoud isolation by chromatography and structure elucidation by chemical and spectral analyses. RESULTS: Five compounds were obtained and elucidated as: 8(E)-N-(2' hydroxypalmityl)-1-O-beta-gly-copyranosyl-3-hydroxyl-9-methyl-2- octodecanine-4, 8-diene (I), N-(2'-hydroxytetracosanoyl)-1,3,4-trihydroxy-2-octodecanine(II), 7, 22-diene-3-hydroxy-6,9-epidioxyergosta(III), ergostol(IV) and alpha-palmitin(V). CONCLUSION: I, II, III were obtained from Gliocladium sp. for the first time. PMID- 12528523 TI - [Studies on difference between sporoderm-broken and nonbroken spores of Ganoderma lucidum (Leyss. ex Fr.) Karst. by polysaccharide analysis]. AB - OBJECTIVE: To compare the release ability of water-soluble polysaccharides in sporoderm-broken and nonbroken spores of Ganoderma lucidum, and establish a comparatively correct method for the determination and analysis of polysaccharide contents in Chinese herbs. METHOD: The release ability of water-soluble polysaccharides was determined on the basis of phenol-sulfuric acid modification in different conditions. RESULT: The release ability of polysaccharides of sporoderm-broken spores was much greater than that of nonbroken spores; and the phenol-sulfuric acid modified cation method proved excellent in accuracy and reproducibility, with a relative error less than 1.5%. CONCLUSION: The spores should be wall-wracked if used as a nutriment, or for extraction and analysis of their effective components. The method can be successfully used for the determination of polysaccharide contents in Chinese herbs or nutriments. PMID- 12528524 TI - [Effects of ginkgo biloba extract on somatosensory evoked potential and nitric oxide after subarachnoid hemorrhage]. AB - OBJECTIVE: To observe the changes of somatosensory evoked potential(SEP) and nitric oxide (NO) after subarachnoid hemorrhage(SAH), and the influence of Ginkgo biloba extract (EGb). METHOD: Rats in sham-operated group, SAH model group and EGb-treated group underwent measurement of dynamic changes of regional cerebral blood flow(rCBF), SEP and NO levels both in serum and in brain tissue within 24 h after operation. RESULT: In SAH group, rCBF decreased immediately after operation, with no tendency to recover within 24 h. The latency of SEP delayed progressively from 1 h to 24 h after SAH. NO levels in serum and in brain tissue decreased and increased respectively from 1 h to 24 h after SAH. EGb effectively antagonized the changes of above parameters. CONCLUSION: SEP is helpful in the judgement on brain ischemic damage after SAH. Decrease of NO in serum and increase of that in brain tissue may lead to cerebral vasospasm and ischemic brain damage respectively after SAH. EGb relieves SAH-induced brain ischemic damage by reversing the pathological alterations of NO. PMID- 12528525 TI - [Effect of Curcuma zedoaria (Berg.) Bosc on the myoelectric activity of uterus in rats and study of its mechanisms]. AB - OBJECTIVE: To investigate the effect of Curcuma zedoaria on the myoelectric activity of uterus in virgin rats and study its mech anisms. METHOD: A pair of bipolar Ag-AgCl electrodes were implanted on the serosal surface of uterus in rats to observe the effect of C. zedoaria on the myoelectric activity of uterus before and after the five agonists were injected intraperitoneally. RESULT: Decoction of C. zedoaria significantly increases the spike area, the duration and the number of bursts of action potentials of the uterine smooth muscle and its effect is related dosage. Atropine and phentolamine decreased the exciting effect of C. zedoaria, whereas verapamil, diphenhydramine and indomethacin have no effect on the excitation of C. zedoaria. CONCLUSION: C. zedoaria has obvious exciting effect on the smooth muscle of uterus in rats, and its mechanisms may be associated with M-receptor and alpha-receptor. PMID- 12528526 TI - [Systematic study on the authentic and superior medicinal herba and GAP of herba asari]. AB - OBJECTIVE: To investigate the methodology of the systematic study on the Authentic and Superior Medicinal Herba and GAP of Herba Asari. METHOD: The study was made by textual criticism of Herbology, Botany, Palynology, Cytology, Chemistry, Pharmacology, the analytic methods of allozyme and RAPD. Many subjects on the herb were investigated, such as history, botanical origin, routine examination, morphology of pollen, chromosome and karyotype, the content of oil and its pharmacological effects and so on. RESULT AND CONCLUSION: The methodology of systematic study on the Authentic and Superior Medicinal Herba was provided. According to the results of the systematic study, the GAP of the herb can be made. PMID- 12528527 TI - [The expression of tyrosine-kinase receptor flk-1 and hypoxic pulmonary vascular remodeling in the lung of rats exposed to hypoxia]. AB - OBJECTIVE: To assess the role of tyrosine-kinase receptor flk-1 in the development of hypoxic pulmonary vascular remodeling. METHODS: We divided 20 male Wistar rats into two groups(control vs hypoxia) and exposed them to normoxic condition and isobaric hypoxia for 3 weeks respectively. The pulmonary artery pressure was measured by right cardiac catheterization. The expression of flk-1 in lung tissues was measured by immunohistochemical staining. Histologic sections of the lungs were examined by a computerized image analyser. RESULTS: In hypoxic rats, the pulmonary artery pressure was significantly raised to a higher level, P < 0.01; the cell number of vascular wall was significantly increased. The results also demonstrated that chronic hypoxia brought about the significant increment in thickness of wall with narrowing of lumen of pulmonary arterioles, and the increment in the percent of vascular wall thickness/vascular external diameter (WT%) and the percent of vascular wall area/total vascular area (WA%), P < 0.01. The positive staining of flk-1 in the wall of pulmonary arteriole of rats treated with hypoxia was significantly stronger than that of normal rats, P < 0.01. Positive correlations were seen between the increment of expression of flk-1 with WT% and WA% (r = 0.714, 0.738, P < 0.01). CONCLUSION: Chronic hypoxia can induce an increasing expression of flk-1, and the flk-1 may play an important role in the pathogenesis of hypoxic pulmonary vascular remodeling. PMID- 12528528 TI - [Study of deletion and mutation of p16 gene in primary hepatocellular carcinoma]. AB - OBJECTIVE: To investigate the role the deletion and mutation of p16 gene plays in the pathogenesis of human primary hepatocarcinoma. METHODS: Thirty-one cases of human hepatocarcinoma, 31 cases of adjacent noncancerous liver cirrhosis and the leukocytes of 8 normal human subjects were analyzed for deletion and mutation in p16 gene exons 1, 2 and introns 1, 2 with comparative multiple PCR and PCR-SSCP. RESULTS: Deletion of p16 gene exon 1 and partial intron 1 was found in 4 of 31 cases (13/%). No deletion of exon 2 or intron 2 was found. Three patterns of p16 gene intron 1 and 18 bp-flanking sequence in exon 2 at SSCP analysis were observed in hepatocellular carcinoma and corresponding adjacent noncancerous cirrhosis, and two patterns were found in human normal leukocyte DNA. No aberrant single strand at SSCP in p16 gene exon 1 or most part of exon 2 or intron 2 was detected. CONCLUSION: Low frequency of deletion and rare mutation of p16 suppressor gene occurred in hepatocellular carcinoma. PMID- 12528529 TI - [Expression of HLA-I antigens in primary hepatocellular carcinoma]. AB - OBJECTIVE: To find out whether hepatocellular carcinoma (HCC) cells express HLA-I antigens. METHODS: LSAB assay of immunohistochemistry was used to detect six frozen HCC specimens. RESULTS: All the HCC cells have strong expression of HLA-I antigens. CONCLUSION: In view of these data on the expression of HLA-I antigens, the authors deem it practical to develop HCC CTL vaccine. PMID- 12528530 TI - [Differential expression of cyclins D1, E and A in human breast diseases]. AB - OBJECTIVE: To understand with greater clearness the effect of overexpression of cyclins gene and the potential implications of it for tumorgenesis in breast. METHODS: We assayed the expression of cyclins D1, E and A in different types of human breast diseases by immunohistochemical staining. RESULTS: Significant difference was seen among the malignant tumor, benign tumor and dysplasis(P < 0.001). The expressive intensity of above three kind of cyclins in malignant tumor was the highest, that in benign tumor was higher, and that in dysplasis was low. CONCLUSION: These data suggest that the expressive intensity of cyclins gene may serve as an indicator for malignant intensity of tumors. The expressive ratio difference of cyclins E and A between infiltrating ductal carcinoma and infiltrating lobular carcinoma may imply that there are different mechanisms involving the occurrence of different histological types of breast tumor. PMID- 12528531 TI - [Study of quinolones resistance in Escherichia coli, Staphylococcus aureus and Pseudomonas areuginosa]. AB - OBJECTIVE: To investigate the factors of quinolone-resistance mediating in Escherichia coli(E. coli), Staphylococcus aureus (S. aureus) and Pseudomonas areuginosa(P. areuginosa) and examine the characteristic changes and quinolones resistance of these bacteria after exposing them to quinolones, dye-materials, and ultraviolet respectively. METHODS: The agar plate inoculating on surface and tube broth two-fold dilution methods were adopted. RESULTS: Quinolone-resistance was mainly mediated by repeated exposure of the bacteria to low level concentration of quinolone. No quinolone-resistance was found after exposure of the bacteria to ultra-violet and dye materials such as Ethidium bromide and Acry orange. Among the three kinds of bacteria, E. coli tended to have stable high level of quinolone- resistance(MICs > or = 256 mg/L). S. aureus acquired mediate level of quinolone-resistance(MICs < or = 32 mg/L). P. aeruginosa acquired a high but not stable level(MICs > or = 256 mg/L) of resistance. These bacteria were cultured in the concentration of 1/2 x MIC, 1 x MIC, 2 x MIC, 4 x MIC, 5 x MIC ciprofloxacin, and were compared with those cultured in the same concentration of sparfloxacin and cefotaxime. After being affected by ciprofloxacin, the E. coli underwent apparent morphologic changes, such as becoming wider(2-3 times), longer(10-30 times), thread-like, and fewer in number. These changes were consistent with the increase of quinolone concentrations. The morphologic changes in P. aeruginosa and S. aureus were not so obvious as those in E. coli. Sparfloxacin and cefotaxime mainly led to the decrease in the numbers of bacteria. When the concentration of Sparfloxacin and cefotaxime were 4 x MICs, the changes of P. aeruginosa and S. aureus were less obvious. CONCLUSION: The bacteria's quinolone-resistance is mainly induced by the quinolone itself. The quinolone-resistance is more likely occur in E. coli than in P. aeruginosa and S. aureus. After exposure of the bacteria to ciprofloxacin, E. coli might have most apparent morphologic changes and occurrence resistance. PMID- 12528533 TI - [Study on the content of serum HDL subclasses in obese subjects]. AB - OBJECTIVE: To investigate the change of the contents of serum HDL subclasses in obese subjects. METHODS: Contents of serum HDL subclasses in healthy controls(n = 64) and obese subjects(n = 58) were determined by two-dimensional gel electrophoresis associated with immunodetection method. RESULTS: The pre-beta 1 HDL, HDL3c and HDL3b contents increased significantly (P < 0.001) and the HDL2b content decreased significantly (P < 0.001) in obese subjects as compared with those in healthy controls. In male obese subjects, the pre-beta 1 HDL and HDL3c levels were significantly higher(P < 0.01 and P < 0.05, respectively) and HDL2b level was significantly lower(P < 0.05), compared with those in female obese subjects. In male control subjects, the level of pre-beta 1 HDL was significantly higher than that in female subjects. In obese subjects, the concentrations of BMI and serum TG had positive correlation with those of pre-beta 1 HDL(r = 0.382, P < 0.001), HDL3c(r = 0.232, P < 0.05) and HDL3b(r = 0.294, P < 0.01) and negative correlation with that of HDL2b(r = -0.304, P < 0.01). CONCLUSION: The particle size of HDL in obese subjects showed a general shift toward smaller size, which indicated that the maturation of HDL could be abnormal in obese subjects. The particle size of HDL was found to be smaller in both obese and control male subjects than in female subjects, suggesting that sex seems to be related to the contents of serum HDL subclasses. PMID- 12528532 TI - [Effects of IL-8 on neutrophils under fluid shear]. AB - OBJECTIVE: In order to elucidate the effect of IL-8 on neutrophils in a systematic and all-round way, we introduced an important physiological environment of neutrophils, fluid flow, into this study on the changes in some responses of neutrophils to IL-8 and the inter relationship of those responses under the conditions of different patterns of flow field. METHODS: After using IL 8 and steady flow or sinusoidal-oscillate fluid shear rate simultaneously acting on separated neutrophils for one minute, we measured the expression of CD18, CD62L and the F-actin content with flow cytometry and measured the free calcium concentration inside cells ([Ca2+]i) with the fluorescent stain Fura2/AM using the fluorescent spectrophotometer. RESULTS: It was found that flow shear stress could influence the expression of surface adherence molecule of neutrophils under the action of IL-8. CD18 surface expression increased, and CD62L shed under different flow shear rates, but no significant relationship of these changes with the intensity and pattern of shear stress was noted. On the other hand, the F actin content in neutrophils under the action of shear stress and IL-8 decreased significantly at low shear rate and then gradually increased at high shear rate (600s -1) to the control value (without shear). The [Ca2+]i concentration under the action of shear stress and IL-8 decreased significantly at low shear rate followed by a quick increase at high shear rate. CONCLUSION: The flow shear stress can influence the activation effects of IL-8 on neutrophils and regulate the magnitude of neutrophil reactions to a great extent, and among these reactions, the change of [Ca2+]i concentration is the most sensitive one and the greatest one. This is in consistence with its second messenger role in the signal transduction pathway. Our results could serve as a basis on which to understand better the mechanism of inflammatory reaction wherein the neutrophils under flow condition of blood are involved. PMID- 12528534 TI - [Expression of Fas and Fas ligand in lesions of patients with discoid lupus erythematosus]. AB - OBJECTIVE: To study the relation of apoptosis and pathogenesis of lupus erythematosus. METHODS: Expressions of Fas and Fas ligand(FasL) were detected in 30 lesions of discoid lupus erythematosus(DLE) and 5 skin specimens from healthy controls by immunohistochemical staining. RESULTS: Fas and FasL were positively expressed in most epidermal keratinocytes, infiltrating lymphocytes around blood vessels and appendages, and Fas was positively expressed in a few sweat ducts and hair follicles. Fas and FasL expressions were weakly positive or negative in healthy controls. CONCLUSION: Apoptosis induced by Fas, FasL pathway may be related to the pathogenesis of lupus erythematosus. PMID- 12528536 TI - [The studies on microtensile bond strength measurements of two dentin adhesives]. AB - OBJECTIVE: The purpose of this study was to investigate the feasibility of a modified microtensile method used to test bond strengths of two current one bottle dentin bond systems (Prime & Bond NT, PBNT; Prime one Mirage, P-One) with a parallel match design. METHODS: 15 extracted, caries-free human molars were cut to expose occlusal dentin. A 5 mm deep slot was prepared in each crown to divide the crown into nearly equal halves for accepting treatment of the two dentin bond systems, respectively. After 24 h storage in distilled water at 37 degrees C, the bonded teeth were subjected to two treatments: 5 teeth were tested without further treatment and 10 teeth were thermocycled (2400 cycles, between 5 degrees C and 55 degrees C) prior to bond strength testing. Hour-glass shaped specimens with a distance of approximately 1.0 mm at the narrowest portion were cut from each tooth and tested in tensile mode. RESULTS: Bond strengths (mean MPa) were: for PBNT: 42 & 31, and for P-One 64 & 38 without and with thermocycling, respectively. Two-way ANOVA revealed a significant difference in bond strengths(P < 0.001) between the two systems and when thermocycled. However, a pairwise multiple comparison (Tukey test) showed that after thermocycling the difference between the two systems was not significant (P > 0.05). Regression analysis showed that a correlation existed between the two systems' tensile bond strength values grouped by tooth (correlation coefficient r = 0.575, P < 0.05). CONCLUSION: The modified microtensile method with a parallel match design is feasible and suitable for evaluating two different bonding systems or dentin treatments. PMID- 12528535 TI - [Human toll like receptor gene expression in human pulmonary granule epithelial cells and umbilical vein endothelial cells]. AB - OBJECTIVE: To determine whether human Toll like receptors (TLRs) genes are expressed in human lung granule epithelial cells and umbilical vein endothelial cells (UVECs). METHODS: Specific primers for TLR2 and TLR4 were designed and total RNAs were isolated from human monocyte cell line THP-1, human lung granule epithelial cell line SPC-A-1 and the primary culture of human UVECs. RT-PCR and Northern blotting with digoxin labeling were used for the detection of TLR2 and TLR4 mRNA expression. RESULTS: TLR4 mRNA expressed in the three types of cells mentioned above. TLR2 mRNA was detected in THP-1 and UVECs, but not in SPC-A-1 cells. By using DNA sequencing, the RT-PCR products were confirmed to be TLR2 and TLR4 cDNAs. CONCLUSION: This study provided evidence indicating that Toll like receptor genes constitutively express not only in human monocytes but also in human larger vascular endothelial cells and pulmonary granule epithelial cells. PMID- 12528537 TI - [Determination of shrinkage and bending strength of GI-II infiltrated ceramic]. AB - OBJECTIVE: To determine the shrinkage and strength of GI-II Infiltrated Ceramic for providing scientific basis of its clinical use. METHODS: The adobes by slip casting alumina powder of GI-II Infiltrated Ceramic were sintered at 1125 degrees C for 2 hours. The sintered alumina bodies were then divided into two groups and infiltrated by melt glass at 1125 degrees C and 1100 degrees C respectively for 6 hours. The length of the infiltrated ceramic specimens before and after fabrication was determined to calculate shrinkage. Their strength values were obtained by 3-point bending test. RESULTS: The infiltrated ceramic infiltrated at 1125 degrees C was 0.289 +/- 0.155% in shrinkage, 367.7 +/- 76.1 MPa in strength, and 4.79 in Weibull modulus; that infiltrated at 1100 degrees C was 0.253 +/- 0.043%, 377.5 +/- 64.4 MPa, and 5.52, respectively. There was no statistically significant difference between those two groups. But their strength values were statistically high, compared with 300 MPa. CONCLUSION: The shrinkage of GI-II Infiltrated Ceramic could be compensated by brushing spacer on dies and expansion of die stone. Its strength satisfied the requirement for clinical use of all ceramic bridge materials. This kind of high-strength and low-shrinkage ceramic could be used for fabricating all ceramic crown and bridge core. PMID- 12528538 TI - [An experimental study on the implantation of a biomaterial with electro-activity for replacement of hard tissue in bone]. AB - OBJECTIVE: To assess the effects of a piezoelectric biological ceramic on osteogenesis. METHODS: Hydroxyapatite (HA) and piezoelectric biological ceramics (hydroxyapatite and barium titanate, HABT) were implanted in the jawbones of 5 dogs, and for sample collection, the dogs were killed separately at 1 week, 2 weeks, 1 month, 2 months and 3 months after implantation. The samples from a rhesus monkey and a blank control were collected 34 months after implantation. The implanted samples and surrounding tissues were subjected to histological observations using light microscopy (LM) and scanning electronmicroscopy (SEM) were made. RESULTS: Compared with the control groups, the HABTs promoted osteogenesis significantly. One week after implantation, new bone tissues were found on the surface vertical to the longitudinal direction of HABT; more bone tissues were found after 2 weeks. HABTs induced the bone tissues to arrange orderly. After two years and ten months of implantation, the piezoelectric bioceramic and bone became monolithic, and the structure of bone was normal. CONCLUSION: HABTs could promote osteogenesis. PMID- 12528539 TI - [A study of bone development and metabolism in childhood]. AB - OBJECTIVE: To explore the regularity of bone development and metabolism in childhood. METHODS: A survey was made to collect and analyze the data on bone biochemistry, the density of lumbar vertebrae and the X-ray examination of calcaneus in 224 healthy children aged 3-15 years in the urban area of Chengdu. RESULTS: Comparative analysis of serum and urine calcium and phosphate concentrations showed no significant difference between the age groups. The serum alkaline phosphatase (ALP) and bone alkaline phosphatase (BALP) were found to increase with age and reach their peak values in children aged 7-12 years, and their values were significantly higher than that of adults (P < 0.05). The density of lumbar vertebrae and the trabecular bone of calcanesus also developed with age. CONCLUSION: The measurements of the bone biochemistry, bone density and the trabecular bone of calcaneus which reflect the bone development and metabolism have their special regularity in childhood. The criteria in use for adulthood is not fit for the evaluation in childhood. The data obtained can be used in clinical diagnosis and treatment of the bone metabolic diseases of children. PMID- 12528540 TI - [Study of peak expiratory flow in normal young, middle-aged and old persons in Chengdu area]. AB - OBJECTIVE: To build the normal ranges and regression equations of peak expiratory flow (PEF) in normal young, middle-aged and old persons. METHODS: PEF, FVC, V75, V50, V25 were measured in 393 normal young, middle-aged and old persons with the youngest aged 20 years and the eldest 84 years. Regression equations of PEF were built for younger group (20 yr-49 yr) and elder group (50 yr-84 yr) respectively. RESULTS: PEF, FVC, V75, V50, V25 were decreased with increase of age. The regression equations showed that both in younger and elder groups, PEF was negatively correlated with age in male (P < 0.0001-0.002, r = 0.32-0.39) and positively correlated with FVC in female (P < 0.0001-0.0005, r = 0.32-0.59). PEF was highly correlated to V75 with r = 0.87-0.94 in male and r = 0.93-0.96 in female. The coefficient of variation (CV) of PEF was in normal range (16%-24%) for all age groups except male and female above 70 years old with CV of 30% and 27% respectively. This confirmed the data was reliable. CONCLUSION: The normal ranges and regression equations of PEF constructed as a result of this study for normal humans of 20-84 years old can be put to use in clinical work and public health. PMID- 12528541 TI - [A comparison of the kinetic characters of NT-3, NT-4 and BDNF retrogradely transported in facial nerve]. AB - OBJECTIVE: To compare the kinetic characters of retrograde transport of neurotrophin-3 (NT-3), neurotrophin-4 (NT-4) and brain-derived neurotrophin factor (BDNF) in facial nerve. METHODS: Radioactive tracer technique was used. After one lateral facial nerve trunk of adult rabbit was transected, a silicone chamber was inserted between the stumps, and 3.7 MBq of 125I-NT-3 or 125I-NT-4 or 125I-BDNF or 125I-HSA was administered into the chamber. At different time-points after injection, the facial nerve trunk and facial nerve motor neurone of brain stem were collected and the uptake rates were measured. The kinetic parameters of each labeled compound were calculated using 3P87 program of kinetics. RESULTS: The transport amount of neurotrophin retrogradely transported by facial nerve is NT-3 > BDNF > NT-4 (P < 0.05), the transport rate is NT-4 > NT-3 > BDNF (P < 0.05). CONCLUSION: The findings could serve as the kinetic characters of retrograde transport of neurotrophins. PMID- 12528542 TI - [The binding characteristics of insulin-MTX to insulin receptor]. AB - OBJECTIVE: It has been reported that several kinds of tumors express increased insulin receptor and the molecules of insulin can be internalized in cells and may thence enter into the nuclei mediated by insulin receptor. In this study, we investigated the receptor binding characteristics of insulin-MTX for the possibility of using insulin as a carrier for carcinoma targeted therapy by receptor mediation. METHODS: MTX(methotrexate) was covalently linked to insulin directly. The insulin-MTX conjugate was purified by polyacrylamine agarose gel electrophoresis and analysed by high performance liquid chromatography and SDS- polyacrylamine agarose gel electrophoresis. Histologically confirmed human hepatocellular carcinoma specimens were obtained from patients at surgery and immediately frozen under -80 degrees C. Cell membrane fractions were isolated by sucrose density gradient centrifugation. Competitive displacement of 125I-insulin with insulin and insulin-MTX binding to insulin receptor were carried out and the values of IC50 and Ki were calculated so as to observe the characteristics of insulin-MTX binding to insulin receptor. RESULTS: Insulin-MTX competed as effectively as insulin with 125I-insulin for insulin receptor. The values of IC50 and Ki for insulin-MTX were 93.82 +/- 19.32 nmol/L and 91.88 +/- 16.86 nmol/L respectively, while the values of IC50 and Ki for insulin were 5.01 +/- 1.24 nmol/L and 4.85 +/- 1.12 nmol/L respectively. CONCLUSION: Insulin-MTX could bind with insulin receptor with high affinity. The result demonstrates us that there is a possibility of using insulin as a carrier for carcinoma targeted therapy by receptor mediation. PMID- 12528543 TI - [The preparation of norA gene probe in Staphylococcus aureus]. AB - OBJECTIVE: To prepare norA gene probe for studies on the mechanism of resistance to fluoroquinolones mediated by norA gene in S. aureus. METHODS: The Dig-labeled norA gene probe was made by PCR. RESULTS: This method is simple and feasible. We have got a lot of probes in a short time. The sensitivity of norA gene probe is high; the probe is secure, easy to use and can be stored for a long time. CONCLUSION: The probe is applicable to further studies on the mechanism of resistance to fluoroquinolones mediated by norA in S. aureus. PMID- 12528544 TI - [The effect of leukotriene antagonist on the parameters of dose-response curve obtained after methacholine challenge test]. AB - OBJECTIVE: To investigate the effect of leukotriene antagonist (zafirlucast) on the parameters of dose-response curve (DRC) obtained after methacholine challenge test. METHODS: A therapy trial was conducted with the use of zafirlucast in 20 cases of cough variant asthma (CVA) and 20 cases of asthma. Placebo was used in 4 cases of CVA and 10 cases of asthma for comparison with the two therapy groups respectively. Before and after the zafirlucast treatment a series of parameters of DRC were obtained after the methacholine challenge test and were calculated. These included the response threshold (PC35sGaw), the dose-response slope (DRS), the ratio of the area under the curve to the logarithm of the maximal concentration (AUC/1 g[Cmax]), and the maximal response. RESULTS: Zafirlucast significantly increased the PC35sGaw in the CVA and asthma groups. It conspicuously decreased all the parameters of DRS and AUC/1 g[Cmax] except for maximal response in the cases of CVA, and it had no significant effects on all the parameters of DRS and AUC/1 g[Cmax] except for PC35sGaw in the cases of asthma. CONCLUSION: Short-term use of Zafirlucast in asthmatic patients could improve the parameters of dose-response curve obtained after the methacholine challenge test. PMID- 12528545 TI - [Effect of total flavones of Hippophae rhamnoides L. on sympathetic activity in hypertension]. AB - OBJECTIVE: To investigate the effect of total flavones of hippophae rhamnoides L (TFH) on sympathetic nerve activity of essential hypertensive patients and to determine whether TFH possesses inhibitory effect on sympathetic activity after supine isometric exercise. METHODS: Heart rate, blood pressure and plasma catecholamines of the patients were measured before and after treatment. 88 patients were randomly divided into TFH group (n = 35), calcium antagonist nifedipine (n = 33) and verapamil ER (n = 20) group. RESULTS: It was shown after 8 weeks of treatment that TFH did not alter the rest heart rate and plasma catecholamine concentration in the patients (P > 0.05). In contrast, nifedipine produced an increase of noradrenaline and adrenaline level(P < 0.01). Isometric exercise may significantly increase the heart rate, blood pressure and plasma catecholamine concentration in hypertensive patients. In TFH group, exercise did not increase the heart rate, blood pressure and plasma catecholamine concentration after treatment. In nifedipine group, however, the same isometric exercise significantly increased the heart rate, blood pressure and noradrenaline concentration after treatment (P < 0.01). In verapamil group, the plasma catecholamine concentration did not change after treatment, but it was significantly increased after isometric exercise. CONCLUSIONS: The results suggest that TFH does not alter the sympathetic activity in treatment of hypertension and the inhibitory effect of TFH on sympathetic activity after supine isometric exercise may provide clinical benefits. PMID- 12528546 TI - [A comparison between the effects of albuterol and isoproterenol in bronchodilation test]. AB - OBJECTIVE: To compare albuterol with isoproterenol as a bronchodilator in pulmonary function test and their value in the diagnosis and differential diagnosis of asthma. METHODS: Two groups (59 patients with asthma and 37 patients with COPD) were included in this study, and the patients of each group were randomly divided into 2 subgroups. The patients in the 2 subgroups inspired the two drugs in a cross-over by odd or even days. The pulmonary functions were evaluated. RESULTS: The changing rates of pulmonary functions induced by the two drugs in the two groups were positively correlated (P < 0.005), The changing rates of FEV1, MMEF, Raw, sGaw induced by albuterol were higher than the ones induced by isoproterenol in patients with asthma(P < 0.02), however the changing rates of FVC and PEF in asthma cases and all the above parameters in COPD cases were not significantly different between the two drugs(P > 0.10). The comparison of positive case number between asthma and COPD cases induced by both drugs showed significant difference(P < 0.005); especially, when the positive rate of sGaw induced by albuterol in asthma cases reached 98%, the positive rates in COPD cases was generally much lower. CONCLUSION: Bronchodilation test is a valuable diagnostic method for asthma. In this study, albuterol was superior to isoproterenol. The most sensitive index in bronchodilation test was sGaw, then in order, FEV1(92%), MMEF and Raw; the FVC and PEF were relatively insensitive. PMID- 12528547 TI - [Studies on liver cancer induced by non-necrotizing dose of diethylnitrosamine in rats]. AB - OBJECTIVE: To provide an appropriate animal model of liver cancer and sensitive indicator for carcinogen risk assessment. METHODS: The developmental processes of liver cancers in rats initiated by non-necrotizing dose of diethylnitrosamine and promoted by phenobarbital were studied. RESULTS: Most of the preneoplastic lesions and liver cancers were made up of eosinophilic cells. The placental glutathione S-transferase (P-GST) staining could allow a lot of positive single cells, foci and nodules to be seen. But the lesions showed by HE staining expressed higher proliferating cell nuclear antigen (PCNA) than those showed by P GST staining expressed, indicating a greater possibility for these lesions to persist and develop further. The cellular morphologic analysis demonstrated that the ratio of nuclear radii to cytoplasm radii decreased in the cell in PCNA positive expressing nodules. In addition, the nodules showed by P-GST expressed higher PCNA than those the foci expressed, indicating an increased possibility to develop further in the nodules. CONCLUSION: PCNA expression may be used as a relatively ideal marker for predicting the potential development of precancerous lesions in the experimental model of rat liver cancer induction. PMID- 12528548 TI - [A study on the relationship between respiratory tract viruses antigen in peripheral blood mononuclear cells and their antibody from children with steroid sensitive and simple nephrotic syndrome]. AB - OBJECTIVE: To explore whether respiratory tract viruses infection was involved in the pathogenesis of sterioid-senstive and simple nephrotic syndrome(SSSNS). METHODS: Using APAAP and ELISA, the authors assayed the expression of respiratory tract viruses antigen in peripheral blood mononuclear cells (PBMC) and the production of viruses antibody in serum from 20 cases of children with steroid sensitive and simple nephrotic syndrome. RESULTS: The positive rate of viruses in the active stage group of SSSNS children increased (P < 0.05), compared with that in the remission group and the healthy control group. The positive rate of virus detection in SSSNS children was closely related with the quantity of urinary protein(P < 0.05). The positive rate of virus detection in the group with urinary protein > or = 50 mg/(kg.d) was higher than that in the group with urinary protein < 50 mg/(kg.d). CONCLUSION: The onset of SSSNS may be related to the infection of respiratory tract viruses. PMID- 12528549 TI - [Inhibitory effect of antisense c-myb cDNA on the growth of asynchronous glioma cells in vivo]. AB - OBJECTIVE: To examine the therapeutic effect of oncogene c-myb antisense oligodeoxynucleotide (AON) on rat C6 glioma in vivo. METHODS: C6 glioma cells were implanted into the subcutis of nude mice, the animals were randomly divided into three groups (12 mice respectively) when the tumor grew about 10 mm in size, and the mice with subcutaneous tumor foci were treated with sense oligodeoxynucleotide (SON), AON and normal saline. Three animals of each group were killed after treatment at 4, 8, 12 and 16 days respectively. The dynamic growth manifestations, features, histopathological changes and apoptosis of the glioma in each group were observed. RESULTS: AON suppressed the growth of C6 glioma cells, the suppression rates being 66.7%, 71.4%, 72% and 73% at 4, 8, 12, 16 days respectively (P < 0.05). The changes in weight were concurrent with the variations of tumor volume, the tumor weight of AON groups was significantly decreased (P < 0.05). The immunohistochemical assay showed the expression of c myb proto-oncogenes was significantly decreased in AON groups (P < 0.05). The flow cytometry analysis found the apoptosis cell percentage of AON groups to be 13.4%, 27.1%, 46.1% and 48.4% at 4, 8, 12 and 16 days respectively, and this percentage of AON groups was about 3.5 times the apoptosis cell percentage of SON and control groups at the corresponding time. In addition, noticeable inflammatory infiltration and calcification were observed after the treatment with AON. CONCLUSION: The above findings indicate that c-myb gene might be associated with the suppression of carcinogenesis in brain glioma, and oncogene c myb might be chosen as a target for antisense gene therapy of gliomas. PMID- 12528550 TI - [The prognostic value of vascular endothelial growth factor in breast cancer]. AB - OBJECTIVE: To define the relevance of VEGF status to the prognosis of breast cancer. METHODS: The tissues of 70 invasive ductal breast carcinomas were evaluated for VEGF protein expression by immunohistochemistry LSAB method, and the prognostic value of VEGF status was assessed in combination with the follow up data on those breast cancer patients. RESULTS: VEGF status was inversely correlated with relapse-free survival and overall survival. Kaplan-Merier survival curve showed that VEGF-rich tumor group had higher relapse rate and mortality rate. Multivariate regression analysis confirmed that the VEGF status had certain prognostic value, but it could not be considered as an independent prognostic indicator. CONCLUSION: VEGF status is one of the significant prognostic indicators of breast cancer. PMID- 12528551 TI - [Changes serum level of tumor specific growth factor and their clinical significance in mechanical trauma patients]. AB - OBJECTIVE: To investigate the post-traumatic changes in the serum level of tumor specific growth factor (TSGF) and their relationship with Injury Severity Score and to inquire into the possible mechanism and clinical significance. METHODS: Fifty patients with mechanical trauma were eligible and included. According to the standard of Injury Severity Score (ISS), the patients were divided into there groups: mild degree group, (ISS < or = 16, n = 31) severe degree group (16 < ISS < 25, n = 13) extreme severe degree group (ISS > or = 25, n = 6). The blood samples of all patients were obtained within 24 hours, and those of 25 dynamically observed patients were obtained on days 3, 5, 7, 9 and 12. RESULTS: The serum level of TSGF was significantly higher in patients with trauma in different groups (P < 0.05). The dynamical observation showed that the index demonstrated curve changes with increase in early stage and decrease later. the climax of TSGF appeared on 7 day in all 3 groups. The TSGF serum levels in extremely severe degree and severe degree groups > mild degree group on days 5, 7, 9 and 12 day (P < 0.05). CONCLUSION: TSGF has relationship with the inflammatory response and recovery of trauma. PMID- 12528552 TI - [The effects of thrombopoietin and interleukin-11 on bone marrow megakaryocytic progenitors in patients with chronic idiopathic thrombocytopenic purpura in vitro]. AB - OBJECTIVE: To observe the effects of recombinant human thrombopoietin (rhTPO) and rhTPO in combination with recombinant human interleukin-11(rhIL-11) on the megakaryocyte (MK) colony growth and maturation in patients with chronic idiopathic thrombocytopenic purpura (CITP) in vitro. METHODS: Bone marrow (BM) mononuclear cells of 21 patients with CITP were cultured in plasma clot culture systems, to which were added rhTPO alone or rhTPO plus rhIL-11 over a range of concentrations. After 14 days, the MK colonies were stained by GP III a Mc Ab(SZ 21) and ABC-Kit and counted. The diameters and areas of the positive cells of GP III a were measured by BM-cell analysis system of MCDS-2010. RESULTS: The diameters and areas of MK in CITP group were significantly lower than that in control group, P < 0.05. The addition of rhTPO to culture systems increased CFU MK, total colonies and diameters and areas of MK in CITP patients. And this promoting effect was not dose-dependent. The optimal concentration of rhTPO was 10 ng/ml; in the group where 10 ng/ml rhTPO was used in combination with 20 ng/ml rhIL-11, the counts of CFU-MK, total colonies and the measurements of areas and diameters of MK increased significantly, compared with those in the group where rhTPO was used alone. CONCLUSION: There are maturation dysfunctions of MK progenitors in patients with CITP. rhTPO alone or in combination with rhIL-11 can promote the proliferation and maturation of MK progenitors in patients with CITP and, moreover, the combination of the two cytokines has more significant promoting effects, compared to rhTPO alone. PMID- 12528553 TI - [Study on psychoprophylaxis and monoamines neurotransmitter of patients with burning mouth syndrome]. AB - OBJECTIVE: Burning mouth syndrome (BMS) is a chronic ache disease, usually occurring in middle aged and old women. This study sought to understand the psychopathologic aspect and monoamines neurotransmitters in the plasma of the patients with BMS. METHODS: Thirty cases were selected (26 females, 4 males); 30 normal control subjects were similar to the BMS cases on age and sex. All subjects were required to complete the Eysenck personality questionnaire (EPQ), and the Self-report Symptom Inventory, Symptom Check List-90 (SCL-90) questionnaire. In case a subject's L (lie) score exceeded 50, she (he) would be removed from the test. 2 ml of blood was drawn from the subject under restine conditions with a fast in the morning to examine norepinephrine and epinephrine contents by high efficient liquid chromatography. Chi-square test, analysis of variance and t'-test were performed. RESULTS: The BMS group had higher scores of nervousness (N) and poikilergasia (P) and lower score of extro/introversion (E) as compared with the control (P < 0.05). The personality types in BMS group were focused on introversion and instability, but in the control group the types were focused on extroversion and stability (P < 0.05). The scores of 9 emotional factors of BMS group were significantly higher than those of the control group (P < 0.05), which indicated that the BMS patients had suffered from serial psychic disorders. The level of plasma norepinephrine in the BMS patients was higher than that of the control (P < 0.01). CONCLUSION: The personality of BMS patients raised body response to harmful stimulations, and obvious psychic disorders in the patient may cause the functional disorders in central and sympathetic nervous systems, which may be associated with BMS' occurrence. PMID- 12528554 TI - [Purification of conjugated bilirubin from human urine]. AB - OBJECTIVES: To develop a simple, practical method with fine yield and purity for separating conjugated bilirubin(Bc) from a sustainable and facile material in order that it could serve as a calibrator for total and direct bilirubin determination. METHODS: Isolating Bc from urine of patients with obstructive jaundice by CHCl3: CH3CH2OH(1:1, V/V) followed by CNBr activated Sepharose 4B-HAS affinity chromatography, we analyzed the products by the use of Doumas J-G diazo method and HPLC, respectively. RESULTS: Determined by J-G diazo method, the crude product extracted by CHCl3: CH3CH2OH(1:1, V/V) yields 90.2 +/- 5.4%(n = 3) of urine Bc, the ratio of Bc to total dry weight being 67.9 +/- 6.4%(n = 3). The final product purified by affinity chromatography with the ratio of separation of 1.5, 1.737 and analyzed by HPLC has the yield of 46.9% and purity of 94.7%(BDG of 76.2%, BMG of 18.5%). CONCLUSION: The above data demonstrate that urine conjugated bilirubin can be successfully purified by CHCl3: CH3CH2OH extraction followed by Sephrose-HSA affinity chromatography with sufficient purity and output. PMID- 12528555 TI - [Measurement of central respiratory drive and inspiratory muscles strength in patients with uremia]. AB - OBJECTIVE: To investigate the changes of central respiratory drive and inspiratory muscles function in patients with uremia. METHODS: We performed the measurement of forced vital capacity (FVC), maximal voluntary ventilation (MBC), forced expiratory volume in one second (FEV1), peak expiratory flow (PEF), maximal midexpiratory flow (MMEF), expiratory flow of 25 per cent of FVC (V25), lung carbon monoxide diffusing capacity (DLco), maximal inspiratory mouth pressure (MIP) and airway occlusion pressure (P0.1) in 25 patients with uremia and 20 normal subjects. RESULTS: In patients with uremia, the FVC, MBC, FEV1, PEF, MMEF and V25 which reflect the lung ventilatory function and the DLco which reflects the lung diffusing function were significantly lower than those in normal subjects. The patients' MIP which reflects inspiratory muscles strength was significantly lower and their P0.1 which reflects the central respiratory drive was significantly higher, compared with the normal subjects'. CONCLUSION: Our findings suggest that on the basis of the disorder of ventilation and diffusing function of the lungs, the inspiratory muscles function of the patients with uremia is significantly decreased and the central respiratory drive of the patients is increased. PMID- 12528556 TI - [Clinical comparison between intraoperative local arterial infusion chemotherapy and systemic venous chemotherapy of gastric cancer]. AB - OBJECTIVE: To evaluate the effect of intraoperative local arterial infusion chemotherapy for gastric cancer. METHODS: The effects of intraoperative local arterial infusion chemotherapy in 40 patients with gastric cancer were assessed. Twenty-four patients with gastric cancers who underwent the intraoperative systemic chemotherapy were taken as controls. The removed specimens were examined microscopically to show the degeneration and necrosis of the tumour cell. RESULTS: After the treatment of the tumour cells, all of the cases in the study group showed various degrees of degeneration and necrosis. The immediate effective rate was 95% for the the study group but only 41.67% for the control group (P < 0.05). CONCLUSION: Intraoperative local arterial infusion chemotherapy for gastric cancer has good immediate effect and little side effect; it prevents the intraoperative iatrogenic metastases and implantation and decreases the postoperative recurrence of gastric cancer. PMID- 12528557 TI - [The effect of salvia miltiorrhiza and shengmai on inflammatory mediator and renal function of post-operative patients with obstructive jaundice]. AB - OBJECTIVE: To investigate the effect of salvia miltiorrhiza and shengmai (SS) on inflammatory mediator and renal function in patients with obstructive jaundice (OJ). METHODS: This study included three groups: the OJ patients receiving SS for 6 days after operation (the SS group, n = 15); the OJ patients not receiving SS (the OJ group, n = 15); the patients with other hepato-biliary disorders (the Non OJ group, n = 15). The levels of ET, LPs, TNF-alpha, IL-6, IL-8 and urine RBP, TFR, ALB were measured one day before operation and 1, 4, 7 days after operation. RESULTS: The SS group's LPs, TNF-alpha, IL-6, IL-8, ET, RBP, TFR and ALB levels were obviously lower on Day 7 after operation as compared with those on Day 1 after operation (P < 0.05). The urine RBP and ALB levels were significantly lower in the SS group than in the OJ group (P < 0.05). CONCLUSION: The post-operative use of salvia miltiorrhiza and shengmai for patients with obstructive jaundice may effectively decrease the postoperative levels of plasma LPs, ET, TNF-alpha, IL-6 and IL-8. At the same time, salvia miltiorrhiza and shengmai may protect renal function by inhibiting inflammatory mediator and improving blood dynamics. PMID- 12528558 TI - [Effect of Astragalus membranaceus injection on megakaryocyte hematopoiesis in anemic mice]. AB - OBJECTIVE: To assess the effect of Astragalus membranaceus injection (AMI) on megakaryocyte hematopoiesis in anemic mice and explore its mechanism. METHODS: Anemic models of mice were randomly divided into two groups: treatment group and anemic control group. Intraperitoneal doses of AMI (20 mg/(ml.20 g) q.d x 6) were given to the treatment group, and equal doses of physiological saline were given to the anemic control group. On days 8, 11 and 14 after treatment, blood platelet and bone marrow cells were determined, and the numbers of CFU-Meg (colony forming unit-megakaryocyte) and Meg-CSA (megakaryocyte colony-stimulating activity) were determined by using technique of hematopoietic progenitor cells culture in vitro. RESULTS: Serum Meg-CSA of the treatment group was significantly higher than that of the anemic control group. The abovementioned indices of the treatment group recovered to normal by day 11, which was markedly earlier than the day of recovery observed in the anemic control group. CONCLUSION: AMI can increase serum Meg-CSA of anemic mice and accelerate the recovery of megekaryocyte hematopoiesis after bone marrow suppression. PMID- 12528559 TI - [Effect of danggui buxue decoction on proliferation and expression of intercellular adhesion molecule-1 in human umbilical vein endothelial cells]. AB - OBJECTIVE: To investigate the effect of Danggui Buxue Decoction (DBD) on proliferation and expression of intercellular adhesion molecule-1 (ICAM-1) in human umbilical vein endothelial cells (HUVECs). METHODS: After treating HUVECs with various concentrations of DBD, we analyzed the proliferation phase of HUVECs and the expression of ICAM-1 using flow cytometry. RESULTS: It was found that DBD could promote the proliferation of HUVECs and the expression of ICAM-1. The expression of CD54 (ICAM-1) was significantly increased in the experiment groups as compared with that in the control group (P < 0.05). In addition, cell cycle analysis revealed more synthetic period cells in the experiment groups than in control group (P < 0.05). CONCLUSION: DBD can promote the proliferation of HUVECs and Increase the expression ICAM-1. PMID- 12528560 TI - [Mitochondrial DNA mutations in matrilineal nonsyndromic deafness pedigrees of southwest China]. AB - OBJECTIVE: To identify the incidence of the 1555A-->G mutation and explore the audiological features of pedigrees with matrilineal non-syndromic deafness in Southwest of China so as to provide the theoretical evidence for establishing the method of gene diagnosis. METHODS: Six pedigrees with 102 members were evaluated audiologically and clinically. DNA was extracted from their blood samples. All subjects were screened for mitochondrial DNA 1555A-->G mutation by Alw 26I restriction endonuclease digestion. RESULTS: Seventeen maternal relatives of aminoglycoside antibiotic induced deafness (AAID) pedigree 1 and pedigree 2, carried 1555A-->G mutation. 10 maternal relatives of Non-AAID pedigree 6 also carried 1555A-->G mutation. No mutation was found among paternal relatives and pedigrees 3, 4 and 5. CONCLUSION: The same audiological features of these pedigrees are: bilateral and symmetrical progressive sensorineural hearing loss with variable age of onset. The 1555A-->G mitochondrial mutation is one of the hereditary factors for this disorder. 4 Aminoglycoside antibiotic plays an important role in developing deafness. The incidence of the 1555A-->G mutation in AAID and matrilineal non-syndromic deafness pedigrees is fairly high. Screening for mitochondrial 1555A-->G mutation may be of great clinical use fullness. PMID- 12528561 TI - [An experimental study on bone marrow purging in vitro for patients with chronic myelogenous leukemia]. AB - OBJECTIVE: To investigate the effect of IL-2, IFN and bcr-abl AS-ODN on bone marrow purging in vitro for patients with chronic myelogenous leukemia (CML). METHODS: By amplifying in vitro clone forming culture, and purging the bone marrow of 5 CML patients through different combined use of IL-2, IFN and bcr-abl AS-ODN, we compared the effects of 4 combined regimens. RESULTS: It was found that the suppressive effects of the 4th regimen combining IL-2 (800 U/ml), IFN (800 u/ml) and bcr-abl AS-ODN (30 micrograms/ml) on leukemia cell clone forming unit (CFU) were significantly different from the suppressive effects of the other 3 regimens (P < 0.01). With the use of the 4th regimen, the GM-CFU and L-CFU survival rates were 26.91% and 3.49% respectively (P < 0.01). CONCLUSION: The combined regimen of IL-2 (800 U/ml), IFN (800 u/ml) and bcr-abl AS-ODN (30 micrograms/ml) has good effect on selective CML bone marrow purging in vitro and can be used in clinical purging of CML bone marrow. PMID- 12528562 TI - [Annexin V for flow cytometric detection of phosphatidylserine expression on lymphoma cells undergoing apoptosis]. AB - OBJECTIVE: To quantitatively analyze apoptotic and secondary necrotic cells under apoptosis conditions. METHODS: The cells of Burkitt lymphoma cell line Raji were incubated with 1.0 mumol/L dexamethasone (DEX) for 2, 4 and 8 h, then stained with Annexin V-FITC (fluorescein isothiocyanate conjugated) which was used to detect the exposure of phosphatidylserine (PS) on the out membrane resulting from a loss of phospholipid asymmetry in the early stage of apoptosis, and also stained with propidium iodide (PI) which allows the analysis of secondary necrotic cells related with cell membrane and DNA damage, then apoptotic cells was quantified by flow cytometry (FMC). Furthermore, Annexin+/PI- and Annexin+/PI+ cells were sorted by fluoresence-activated cell sorter (FACS), and identified by electron microscopy (EM) and DNA gel electrophoresis. RESULTS: The results revealed that the percentage of apoptotic cells was increased and correlated well with incubation time (r = 0.97). The sensitivity of this method was shown by its detection limit 0.02%; the method was reproducible, and the coefficient variance (CV) was 4.2%. Meanwhile, the Annexin+/PI- and Annexin+/PI+ cells were identified as apoptotic and necrotic cells under EM, and the DNA extracted from the Annexin+/PI- cells was characterized by "ladder pattern". CONCLUSION: Annexin V assay for analyzing apoptotic cells is specific, sensitive, accurate, reproducible and quantitative for apoptosis investigation. PMID- 12528563 TI - [Evaluating wall motion of dilated cardiomyopathy by pulsed-wave Doppler tissue imaging]. AB - OBJECTIVE: To evaluate the characteristics of wall motion in cases of dilated cardiomyopathy (DCM) by the use of Doppler tissue imaging (DTI). METHODS: We observed 30 DCM patients and 30 normal volunteers. For the assessment of short axis motion, the myocardial velocities (MV) and transmural myocardial velocity gradients (MVG) of the middle segments of anterior interventricular septum and left ventricular posterior wall were recorded through parasternal long axis view. We detected the MV of the middle segments of left ventricular walls to evaluate the long axis motion through apical windows(4-chamber, 2-chamber and long axis). RESULTS: Four kinds of abnormal presentations of the waves were seen in DCM cases. The patients' MV of were obviously lower as compared with the controls', especially in long axis; the patients' MVG also decreased significantly (P < 0.05). In the patient group, the heterogeneity of wall motion disappeared too. CONCLUSION: PW-DTI can quantitatively assess the above abnormalities of DCM and provide supplementary information for clinical use. PMID- 12528564 TI - [Application of Ca(2+)-ISME in the studies on pharmacokinetics of calcium supplements]. AB - OBJECTIVE: To determined the concentration of Ca2+ in serum by a self-made Ca(2+) ISME in order to indicate the absorption of calcium supplement. METHODS: We have developed a calibration curve method to measure Ca2+ activities in serum sample. The pharmacokinetics of three kinds of calcium supplements were investigated by the Ca(2+)-ISME after P.O. administration in 5 healthy rats. RESULTS: Though the Tmax (hours) of the three kinds of preparations were different, there were no significant differences among their AUC by paired t-text. CONCLUSION: The above results demonstrate that our self-made Ca(2+)-ISME is applicable to the researches on pharmacokinetics of calcium supplements. PMID- 12528565 TI - [Determination of roxithromycin in serum by high-performance liquid chromatography with ultraviolet detection]. AB - OBJECTIVE: To make better the RP-HPLC method for the determination of roxithromycin(RM) in human serum. METHODS: RM and clarithromycin (internal standard) were extracted from alkalinized serum sample (500 microliters) with methylene chloride. After evaporation of the organic layer, the residue was dissolved in 100 microliters of acetonitrile-ammonium phosphate (1:1, pH 6.0) and washed with n-hexane, then 20 microliters was injected onto a column (5 microns, 15 cm x 4.6 mm) of Penomenex luna C18. The mobile phase was acetonitrile-0.05 mol/L phosphoric acid (39:19:42, adjusted to pH 7.2 with ammonia water) pumped at 1.2 ml/min through the column. The variable wavelength UV detector operated at 0.01 aufs and the wavelength was set at 210 nm. RESULTS: The retention times for RM and clarithromycin were 4.4 min and 5.0 min, respectively. Standard curve was linear in the concentration range of 0.25 to 32 mg/L. The detection limit in serum was 0.06 mg/L; the average method recovery 97.4%; the inter-day RSD less than 3.0%. CONCLUSION: This method was found to be simple, rapid, sensitive and accurate for determination of RM in human serum. PMID- 12528566 TI - [The MRI diagnosis of aortic dissection]. AB - OBJECTIVE: To analyze and address the MRI manifestations and diagnostic importance of aortic dissection. METHODS: The MRI manifestations in 31 cases of aortic dissection were analyzed retrospectively. Sixteen cases were proved by operation and pathology. RESULTS: MRI depicted an intimal flap and a double lumen in all 31 patients. The site of intimal tear was located in 7 cases. Also demonstrated were the complications: aortic dilation in 15 cases, aortic branches involvement in 5, thrombus formation in the false lumen in 5, aortic regurgitation in 10, left ventricle hypertrophy in 10, and leak into thorax in 2. CONCLUSION: As a non-invading imaging method, MRI can demonstrate the site, range and complications of aortic dissection; it is very valuable in the diagnosis of aortic dissection. PMID- 12528567 TI - [Ultrasonic multiple measurement estimation of fetal weight in parturient]. AB - OBJECTIVE: To evaluate the ultrasonic measurements of parturient fetal cerebellar diameter (CD), liver length(LL), kidney length (KL) and femur thigh soft tissue thickness(FSTT) for the estimation of fetal weight. METHODS: Ultrasonic measurements of the parturient fetal cerebellar diameter, liver length, kidney length and femur thigh soft tissue thickness were made in 115 cases. The data on these parameters were analyzed with statistical methods, and a multiple linear regressional equation for predicting the fetal body weight was derived. The calculated value of fetal body weight and the estimated parameters of the fetal body were compared with the actually measured values immediately after birth. The error of estimation was analyzed. RESULTS: CD, LL, KL and FSTT were significantly correlated with neonatal birth weight, the coefficient of multiple correlation R = 0.9425 (P < 0.001); among the 115 cases, 101 had absolute error less than +/- 200 g, with an agreement rate of 88%. The sensitivity and accuracy for detection of normal fetus were 98.13% and 94.78% respectively. The multiple linear regression equation derived was BW = -1009.914 + 356.1303FSTT + 354.0682KL + 236.8484 LL + 179.7384 CD. CONCLUSION: The equation is a simple, accurate and valuable equation for the estimation of fetal weight and can be used in clinical practice. PMID- 12528568 TI - [Gigantism with low serum level of growth hormone: a case report]. AB - Gigantism with low or normal basal concentrations of growth hormone (GH) is a rare condition, possibly due to abnormal GH secretory patterns, enhanced tissue sensitivity to GH, or the existence of an unidentified growth promoting factor. Here we report an 11 year-old female case of gigantism with a normal pituitary gland. Her height was 181 cm, body weight 77 kg, and bone age 11.1 years. Her basal serum GH levels were lower than 1 ng/ml. The levels of T3, T4, FT3, FT4, TSH, E2, LH, FSH, PRL, PTC and ACTH were normal. Serum GH response to insulin induced hypoglycemia or arginine stimulation tests was blunted. In this case, non pulsatile GH secretion and enhanced tissue sensitivity to GH may induce hypersecretion of IGF-1 and the existence of an unidentified growth promoting factor or biologically active anti-GH receptor antibodies may cause clinical gigantism. PMID- 12528569 TI - Microsystems in health care: Part 2. Creating a rich information environment. AB - BACKGROUND: A rich information environment supports the functioning of the small, functional, frontline units--the microsystems--that provide most health care to most people. Three settings represent case examples of how clinical microsystems use data in everyday practice to provide high-quality and cost-effective care. CASES: At The Spine Center at Dartmouth, Lebanon, New Hampshire, a patient value compass, a one-page health status report, is used to determine if the provided care and services are meeting the patient's needs. In Summit, New Jersey, Overlook Hospital's emergency department (ED) uses uses real-time process monitoring on patient care cycle times, quality and productivity indicator tracking, and patient and customer satisfaction tracking. These data streams create an information pool that is actively used in this ED icrosystem--minute by minute, hourly, daily, weekly, and annually--to analyze performance patterns and spot flaws that require action. The Shock Trauma Intensive Care Unit (STRICU), Intermountain Health Care, Salt Lake City, uses a data system to monitor the "wired" patient remotely and share information at any time in real time. Staff can complete shift reports in 10 minutes. DISCUSSION: Information exchange is the interface that connects staff to patients and staff to staff within the microsystem; microsystem to microsystem; and microsystem to macro-organization. PMID- 12528570 TI - Patient Safety Leadership WalkRounds. AB - BACKGROUND: In the WalkRounds concept, a core group, which includes the senior executives and/or vice presidents, conducts weekly visits to different areas of the hospital. The group, joined by one or two nurses in the area and other available staff, asks specific questions about adverse events or near misses and about the factors or systems issues that led to these events. ANALYSIS OF EVENTS: Events in the Walkrounds are entered into a database and classified according to the contributing factors. The data are aggregated by contributing factors and priority scores to highlight the root issues. The priority scores are used to determine QI pilots and make best use of limited resources. Executives are surveyed quarterly about actions they have taken as a direct result of WalkRounds and are asked what they have learned from the rounds. RESULTS: As of September 2002, 47 Patient Safety Leadership WalkRounds visited a total of 48 different areas of the hospital, with 432 individual comments. DISCUSSION: The WalkRounds require not only knowledgeable and invested senior leadership but also a well organized support structure. Quality and safety personnel are needed to collect data and maintain a database of confidential information, evaluate the data from a systems approach, and delineate systems-based actions to improve care delivery. Comments of frontline clinicians and executives suggested that WalkRounds helps educate leadership and frontline staff in patient safety concepts and will lead to cultural changes, as manifested in more open discussion of adverse events and an improved rate of safety-based changes. PMID- 12528571 TI - Deciding not to measure performance: the case of acute otitis media. AB - BACKGROUND: No Health Plan Employer Data and Information Set (HEDIS) performance measures evaluate health plans for possible overuse or inappropriate use of antibiotics. Acute otitis media (AOM), or infection of the middle ear, is one of the most common infections in children. The antibiotic resistance of the bacteria that cause AOM and the general overuse and inappropriate use of antibiotics for this condition are taking center stage as a major public health threat. An effort was undertaken to develop a new HEDIS performance measure that evaluates appropriate antibiotic use in children with AOM. THE MEASURE DEVELOPMENT PROCESS: The measure development process has three major phases: the initial development phase, the field-testing and analysis phase, and the measure refinement phase. With AOM, the measure development process could not proceed beyond the first phase for reasons that are be discussed in detail. CONCLUSIONS: Additional difficulties beyond feasibility issues may arise in developing a quality of care performance measure. The measure development process discussed in this article failed primarily because of issues related to relevance as well as the scientific soundness of the proposed measures. If the evidence base related to the diagnosis and management of a particular condition is sparse, no standard can be developed against which to measure performance, and the criterion of scientific soundness cannot be met. This is the case with AOM. Unfortunately, the evidence base will likely remain inadequate to support the development of such a measure in the future. PMID- 12528572 TI - Right programming of pumps to prevent errors in the infusion process. AB - A problem area emerged in the programming, administration, and monitoring of infusion pumps. The authors describe their strategies for preventing general infusion pump errors and monitoring patient responses. PMID- 12528573 TI - Strategies for addressing the evolving nursing crisis. AB - BACKGROUND: The shortage of registered nurses, which is already having ill effects on the U.S. health care delivery system, is burgeoning at a time when patient acuity is high, care is complex, and demand for services often exceeds capacity. This is a prescription for danger. CREATE A CULTURE OF RETENTION: Some hospitals fare better in recruiting and retaining nurses than others. Top-level managers who provide nurses with delegated authority, adequate staffing, competitive compensation, and a collaborative culture have a built-in resistance to cyclical nursing shortages--theirs is too good a place to work to leave. BOLSTER THE NURSING EDUCATIONAL INFRASTRUCTURE: The shortage of nurses is mirrored by a corresponding shortage of nursing faculty. As nursing educator attrition continues, it is unclear where the future nursing school faculty will come from. Federal funding of nursing education is modest. Structured postgraduate training programs for nurses could provide an opportunity for skill building in real clinical settings. ESTABLISH FINANCIAL INCENTIVES FOR INVESTING IN NURSING: Although there is clearly a business case for creating a culture of nursing staff retention-based on lower turnover, lower costs, higher profitability, and better outcomes--there is just as clear a need for the investment of new dollars in hospitals to establish a new base of response capacity. For hospitals to be truly able to invest in nursing and to resolve the problems that have led to the impending nurse staffing crisis, new federal monies specifically targeted for nursing need to be made available. PMID- 12528574 TI - Making the business case for patient safety. AB - The authors explain why there appears to be a business case for health care organizations to make investments to enhance patient safety. PMID- 12528575 TI - Short-wave diathermy: current clinical and safety practices. AB - BACKGROUND AND PURPOSE: Short-wave diathermy (SWD) is widely available, yet a comprehensive examination of current clinical practice remains absent from the literature. The present paper aims to assess clinical and safety issues in continuous (CSWD) and pulsed (PSWD) short-wave diathermy application and subsequently indicate areas for future research. METHOD: A postal survey was carried out among 116 senior physiotherapists in 41 Irish hospital-based physiotherapy departments. RESULTS: The response rate to the study was 75%. Analysis found that PSWD was the preferred mode of treatment with 27% of respondents using it more than once daily. Respondents considered both modes of treatment indicated for a variety of conditions. CSWD was rated as an effective treatment for chronic osteoarthritis, polyarthritis, non-specific arthrosis and haematomas. PSWD was reported an effective modality for acute soft tissue injury, haematomas, acute osteoarthritis, sinusitis and rheumatoid arthritis. Dose selection varied greatly but tended to be based on the type, nature and duration of the condition. Analysis of safety practices uncovered concerning findings. Although a high level of agreement was found on measures for patient safety, 30% of respondents reported that no measures for operator safety were taken and only five respondents stated they remained a specified distance from SWD equipment. Measures to ensure the safety of other personnel in the physiotherapy department were also lacking. CONCLUSIONS: Given the availability of SWD equipment and its apparent efficacy in certain conditions, future research should aim to establish this by means of controlled clinical trials. The findings on safety practices underline the urgent need for comprehensive guidelines to ensure the safety of operators, patients and the general public during SWD application. PMID- 12528576 TI - Outcomes after trans-tibial amputation: the relationship between quiet stance ability, strength of hip abductor muscles and gait. AB - BACKGROUND AND PURPOSE: Previous research regarding the symmetry of trans-tibial amputees has examined weight distribution and various gait parameters between prosthetic and sound limbs. However, to date, no known research has determined if asymmetry is present in the strength of the hip abductor muscles or if correlations exist between these categories of symmetry. The purpose of the present study was, therefore, to document asymmetry present in stance, strength and gait measures, and to determine the relationship between these variables. METHOD: Twenty-three elderly, unilateral trans-tibial amputees stood on two adjacent forceplates whilst the weight distribution and standard deviation (SD) of the anterior-posterior and the medio-lateral centre of pressure excursion (COPE) under each limb was recorded during four 40 s trials: quiet stance (QS), with eyes open and eyes closed; and even stance (ES), with eyes open and eyes closed. Gait measures (velocity, cadence, step and stride lengths, stance:swing ratio and period of double support) over 10 m of fast, yet safe walking and measures of the strength of hip abductor muscles were also obtained by use of a stride analyser and a dynamometer, respectively. RESULTS: No significant differences were found between QS and ES measures. However, significantly more weight was taken on the sound limb than on the amputated limb. Notably, more anterior-posterior movement occurred under the sound limb than the amputated limb, with this becoming more apparent with the eyes closed. Movement in the medio-lateral direction was found to be the same between sides. No differences in muscle strength or gait measures between limbs were demonstrated. However, strong hip abductor muscles were correlated with increased weight-bearing on the amputated limb, improved gait parameters and reduced medio-lateral COPE under the amputated limb. CONCLUSIONS: This research confirms the asymmetrical nature of amputee stance and demonstrates symmetry of strength and gait measures between limbs. The correlations between hip abductor muscle strength, weight distribution and gait measures illustrates the importance of pre- and post-operative training of these muscles. PMID- 12528577 TI - Low back pain in individuals with chronic airflow limitation and their partners- a preliminary prevalence study. AB - BACKGROUND AND PURPOSE: The prevalence of low back pain in individuals with chronic airflow limitation, and their partners, is unknown. Individuals with chronic airflow limitation, and their respective partners, are likely to have several risk factors that may increase the predisposition for low back pain. The aim of the present study was to provide preliminary data on the prevalence of low back pain in individuals with chronic airflow limitation, and their partners. METHOD: A cross-sectional observational study design was used. Individuals with chronic airflow limitation, and their partners, were approached at South Australian metropolitan Lung Support Group meetings and invited to participate in the study. Three commonly used questionnaires: the Medical Outcomes Survey Short Form 36 (MOS SF-36); the St George's Respiratory Questionnaire (SGRQ); and the Nordic Low Back Pain Questionnaire (NLBPQ) were used to collect data on general health, respiratory health and low back pain prevalence, respectively. Odds ratios and independent Student's t-tests were used to analyse data by use of Epi Info Version 6.0 software. RESULTS: Sixty subjects participated in the study: 41 individuals with chronic airflow limitation and 19 partners of individuals with chronic airflow limitation. The lifetime, 12-month and seven-day prevalence of low back pain in individuals with chronic airflow limitation was 69%, 58% and 31%, respectively. The prevalence of low back pain in the partners of individuals with chronic airflow limitation was found to be higher, at 74%, 68% and 58%, respectively. Four significant relationships between general and respiratory health, and low back pain were demonstrated for individuals with chronic airflow limitation. CONCLUSIONS: The prevalence of low back pain in individuals with chronic airflow limitation was comparable to the lifetime prevalence for the general and older population, and greater than the 12-month prevalence reported for the older population. The prevalence of low back pain for the partners of individuals with chronic airflow limitation was consistently higher than the lifetime and 12-month prevalence reported for individuals with chronic airflow limitation, the older population and family care workers. These findings suggest a larger prevalence study of low back pain in individuals with chronic airflow limitation, and their partners, is warranted. PMID- 12528578 TI - A study of the facilitation of respiration in myotonic dystrophy. AB - BACKGROUND AND PURPOSE: Dystrophia myotonica or myotonic dystrophy is a progressive neuromuscular disorder in which patients demonstrate an irregular respiratory pattern and are particularly subject to cardiopulmonary compromise. The aim of the present study was to investigate the effects of both proprioceptive neuromuscular facilitation (PNF) and staged basal expansion (SBE) breathing exercises in subjects with myotonic dystrophy in two different positions: high support sitting and left side-lying. METHOD: A randomized, double blind study design was used. Seven non-congenital myotonic dystrophy subjects took part in the study. Six 'treatment' levels were applied to each subject: resting in high support sitting; resting in left side-lying; PNF of deep breathing in high support sitting; PNF of deep breathing in left side-lying; SBE in high support sitting and SBE in left side-lying. The outcome measures employed were arterial oxygen saturation (SpO2) and heart rate, as measured by oximetry and thoraco-abdominal motion (TAM), and respiratory rate, as measured by a pneumograph. RESULTS: The PNF technique was found to be the main contributor to improvement in SpO2 for subjects with myotonic dystrophy, where a 2.2% increase was found in the high support sitting position and a 2.6% increase was found in the left side-lying position. There was an increase of between 377% and 556% in TAM during application of both treatment techniques, in either the high support sitting or left side-lying positions. Respiratory rate declined between 15% and 30% immediately after treatment application and heart rate dropped slightly by between 0.2% and 4.1%. CONCLUSION: The present study provides objective evidence that application of these respiratory physiotherapy interventions elicits an improvement in respiratory function in subjects with myotonic dystrophy. Further research into the physiological effects of these techniques could explore the mechanisms responsible for improvement in respiratory indices. PMID- 12528579 TI - Intra-tester reproducibility of pressure biofeedback in measurement of transversus abdominis function. AB - BACKGROUND AND PURPOSE: Strength, co-ordination and timing of transversus abdominis muscle contraction may be important in stabilizing the low back, and contraction of this particular abdominal muscle may be one factor capable of preventing and reducing low back pain in sport and in activities of daily living. The function and strength of the transversus abdominis muscle is difficult to measure. The pressure biofeedback unit (PBU) is a device designed to teach and measure transversus abdominis muscle function. The purpose of the present study was to evaluate intra-tester reproducibility of the PBU. METHOD: The study used a test-retest design. Fifteen healthy subjects (mean age 22.2 years, range 19-28 years) with no history of low back pain participated in the study. After two sessions of practising transversus abdominis muscle contraction, its function was measured in prone position with the PBU device placed over the transversus abdominis muscle. Subjects were instructed to draw in the abdominal wall (for example, lifting it away from the device) after a standardized procedure. Two tests were conducted on two different days, with a seven-day interval in between. RESULTS: Coefficient of variance (CV) was 21.0% and the standard deviation (SD) of delta was 1.59. CONCLUSION: The use of the PBU should be improved for scientific purposes. However, the device may have a role in providing biofeedback to assist in the instruction of correct transversus abdominis muscle contraction. PMID- 12528580 TI - Participating in primary care research. PMID- 12528581 TI - Breast cancer prevention. PMID- 12528582 TI - Understanding laboratory testing in diagnostic uncertainty: a qualitative study in general practice. AB - BACKGROUND: Better knowledge of the professional's motives for ordering laboratory tests in the case of diagnostic uncertainty may lead to interventions directed at reducing unnecessary testing. AIM: To gain insight into the general practitioner's (GP's) motives for ordering laboratory tests for patients presenting with unexplained complaints. DESIGN OF STUDY: Semi-structured interviews based on surgery observations. SETTING: Twenty-one general practices in rural and urban areas of The Netherlands. METHOD: Investigation of the GP's perception of determinants of test-ordering behaviour in the situation of diagnostic uncertainty. The interviews were structured by evaluating the consultations and test-ordering performance of that day. RESULTS: Dutch GPs vary considerably in their motives for ordering tests. Numerous motives emerged from the data. Some examples of important themes include: personal routines; tolerance of diagnostic uncertainty; time pressure; and tactical motives for test ordering. Complying with the perceived needs of the patient for reassurance through testing is seen as an easy, cost- and time-effective strategy. A clear hierarchy in the determinants was not found. CONCLUSION: The decision to request laboratory testing is the result of a complex interaction of considerations that are often conflicting. Designers of interventions meant to improve the ordering of tests should be aware of the numerous determinants, and take contextual variables into account. PMID- 12528583 TI - Qualitative insights into practice time management: does 'patient-centred time' in practice management offer a portal to improved access? AB - BACKGROUND: Different sets of literature suggest how aspects of practice time management can limit access to general practitioner (GP) care. Researchers have not organised this knowledge into a unified framework that can enhance understanding of barriers to, and opportunities for, improved access. AIM: To suggest a framework conceptualising how differences in professional and cultural understanding of practice time management in Auckland, New Zealand, influence access to GP care for children with chronic asthma. DESIGN OF STUDY: A qualitative study involving selective sampling, semi-structured interviews on barriers to access, and a general inductive approach. SETTING: Twenty-nine key informants and ten mothers of children with chronic, moderate to severe asthma and poor access to GP care in Auckland. METHOD: Development of a framework from themes describing barriers associated with, and needs for, practice time management. The themes were independently identified by two authors from transcribed interviews and confirmed through informant checking. Themes from key informant and patient interviews were triangulated with each other and with published literature. RESULTS: The framework distinguishes 'practice-centred time' from 'patient-centred time.' A predominance of 'practice-centred time' and an unmet opportunity for 'patient-centred time' are suggested by the persistence of five barriers to accessing GP care: limited hours of opening; traditional appointment systems; practice intolerance of missed appointments; long waiting times in the practice; and inadequate consultation lengths. None of the barriers is specific to asthmatic children. CONCLUSION: A unified framework was suggested for understanding how the organisation of practice work time can influence access to GP care by groups including asthmatic children. PMID- 12528584 TI - A randomised controlled trial of clinics in secondary schools for adolescents with asthma. AB - AIM: To compare a nurse-led clinic in schools versus care in general practice for adolescents with asthma. DESIGN OF STUDY: Randomised controlled trial in four schools; parallel observational study in two schools. SETTING: Six comprehensive schools. METHOD: In the randomised trial, pupils were invited to attend asthma review at a nurse-led clinic either in school, or in general practice. The parallel observational study compared pupils invited to practice care within and outside the randomised trial. Primary outcome measures were attendance for asthma review, symptom control, and quality of life. Secondary outcomes were knowledge, attitudes, inhaler technique, use of steroids, school absence, peak flow rate, preference for future care, health service utilisation, and costs. RESULTS: School clinic pupils were more likely to attend an asthma review than those randomised to practice care (90.8% versus 51.0% overall [P < 0.001, not consistent across schools]). No differences were observed in symptom control (P = 0.42) or quality of life (P = 0.63). Pupils attending school clinics had greater knowledge of asthma (difference = +0.38, 95% CI = 0.19 to 0.56), more positive attitudes (difference = +0.21, 95% CI = 0.05 to 0.36), and better inhaler technique (P < 0.001, not consistent across all schools). No differences were observed in school absence or peak flow rate. A majority (63%) of those who had received care at school preferred this model in future. Median costs of providing care at school and at the practice were 32.10 Pounds and 19.80 Pounds, respectively. No differences were observed between the groups in the observational comparison on any outcome. CONCLUSIONS: The schools asthma clinic increased uptake of asthma reviews. There were improvements in various process measures, but not in clinical outcomes. PMID- 12528585 TI - Factors associated with the provision of anti-smoking advice in general practice consultations. AB - Guidelines urge general practitioners (GPs) to discuss smoking with patients as frequently as possible. Using data collected before and after consultations, this study confirms that GPs are more likely to discuss smoking in the context of smoking-related problems. Encouraging GPs to make greater use of problem orientated opportunities to discuss smoking may have more effect on rates of advice giving than urging them to advise all smokers. PMID- 12528586 TI - Clinically inexplicable frequent attenders in general practice. AB - This study was based in an isolated rural practice, and it identified 107 frequent attenders (1.95% of the practice population), who created five times the consultation workload of a similar group matched for age and sex. The general practitioners (GPs) classified 60 (56%) of these patients as attending for clinically inexplicable reasons. This subgroup had higher rates of long-term medication and hospital referral, and more free access to primary health care. The study also identified very high levels of kinship and relationship by marriage within this group (47%). PMID- 12528587 TI - The limited utility of the Mini-Mental State Examination in screening people over the age of 75 years for dementia in primary care. AB - The Mini-Mental State Examination (MMSE) is used worldwide to assess cognitive status and it has been recommended for use in primary care to detect dementia. In this study, the MMSE was administered during annual health checks for patients over 75 years of age in nine practices by a member of the primary care team. The mean age was 80 years. Of the 709 patients screened, 286 scored at or below the cut-off point, which was set at 26/30 on the MMSE, and they were invited to be assessed further by a researcher, using the well-validated GMS-AGECAT diagnostic system. Eighty-four of these patients refused, 173 were identified as not having dementia, and 29 (14%) were identified as having dementia. These results, with an 86% false-positive rate, raise concerns regarding the utility of the MMSE as a screening instrument for dementia in primary care. PMID- 12528588 TI - Consultation length, patient-estimated consultation length, and satisfaction with the consultation. AB - Lack of time is a frequently expressed patient concern, but actual measured consultation length is often not associated with patient satisfaction. Correlational analysis of patients from nine GP practices was used to test the hypothesis that patients' perceptions of consultation length are influenced not just by actual consultation length, but by other aspects of their experience of consultations. The consultations of 160 patients were timed, and patients in subsequent interviews gave estimates of consultation duration and completed a satisfaction questionnaire. Consultations where patients were more satisfied appeared to patients to have lasted longer (partial correlation r = 0.26), but were not actually longer. Patient concerns about time may be as much about quality time as about actual time. PMID- 12528589 TI - Supporting ethical practice in primary care research: strategies for action. AB - Researchers in primary care share the general ethical obligations of all researchers. However, these obligations may raise different issues in the unique context of primary care. Current professional ethical guidance for primary care research is complex and fragmentary. The newly introduced research governance framework does not specifically address primary care research, and recent changes in legal requirements have significant implications for research. In this paper, ethical issues arising from research in primary care are considered, current standards and resources are described, and strategies for supporting ethical practice are discussed. Four ethical issues are discussed: consent and competence; confidentiality; power relations; and procedural issues. In the final part of the paper, broad strategies to support ethical practice in primary care research are recommended. These include education and resources, greater clarity of relevant standards, financial support, a greater role for primary care networks, and greater public debate. PMID- 12528591 TI - Educational outreach to improve vaccination uptake. PMID- 12528590 TI - The relationship between consultation length, process and outcomes in general practice: a systematic review. AB - The aim of the study was to examine differences in consultation process and health outcomes between primary care physicians who consult at different rates. A systematic review of observational studies was carried out, restricted to English language journal papers reporting original research or systematic reviews. Qualitative analysis with narrative overview of methodology and key results was undertaken, using MEDLINE (1966 to 1999), EMBASE (1981 to 1999), and the NHS National Research Register. Secondary references from this search were also considered for inclusion. Main outcome measures were objectively measured process or healthcare outcomes. Thirteen papers, describing ten studies, were identified. There were consistent differences in several elements of process and outcome between general practitioners (GPs) who consult at different rates. Although average consultation length may be a marker of other doctor attributes, the evidence suggests that patients seeking help from a doctor who spends more time with them are more likely to have a consultation that includes important elements of care. PMID- 12528592 TI - Educational outreach to improve vaccination uptake. PMID- 12528593 TI - Chronic fatigue syndrome/ME. PMID- 12528594 TI - Health and safety--in general practice? PMID- 12528595 TI - Driving under the influence of insulin. PMID- 12528596 TI - Prescribing antimicrobials for UTI. PMID- 12528597 TI - Warfarin safety. PMID- 12528598 TI - The stupidity of Elizabeth Fry--was it dyslexia? PMID- 12528599 TI - A common pathway: asthma and allergic rhinitis. AB - Historically, the structural and functional differences within the respiratory tract have been the basis for separating the airway into upper and lower entities. For centuries, researchers suggested similar mechanisms in asthma and rhinitis. The implications of such a connection are significant in that asthma and allergic rhinitis (AR) are associated with a high prevalence in adults and children, substantial health care costs, and, often, serious negative effects on quality of life. The importance of the unified airway hypothesis lies in the rational approach to treatment. Management approaches that consider the association between asthma and AR may improve overall outcomes in patients with both diseases. Because the treatment of AR may affect concomitant asthma, significant improvements in health status may occur in some patients. The following review discusses the epidemiology of asthma and AR, provides evidence for common pathophysiological mechanisms, and discusses a therapeutic approach that has positive effects on both diseases, and thereby may maximize benefits and outcomes for patients with concomitant asthma and AR. PMID- 12528600 TI - Airway hyperreactivity: managing outcomes and new directions. AB - Bronchial hyperreactivity is defined as an increased sensitivity to bronchoconstrictor stimuli. It is regarded as the physiological hallmark of bronchial asthma. It is usually associated with a positive family history of asthma, atopy, and evaluated total serum immunoglobulin E. Its prevalence in the normal population ranges from 10 to 20%; it increases with age and is significantly associated with both allergic rhinitis and atopic dermatitis. The relationship between bronchial hyperreactivity and respiratory symptoms may be weak. Bronchial hyperreactivity varies in severity with allergen exposure, viral infection, exposure to pollutants, environmental avoidance measures, anti inflammatory therapy, and immunotherapy. PMID- 12528601 TI - Immunotherapy update: mechanisms of action. AB - Allergen immunotherapy (IT) results in reduction of symptoms of allergic rhinitis and asthma. There still is not satisfactory evidence as to the best marker that explains clinical responses. IT inhibits the early and late nasal, bronchial, and cutaneous responses to allergen challenge. There are increases in antiallergen immunoglobulin G (IgG; 2- to 10-fold) and IgG4 (10- to 100-fold), a gradual decline in antiallergen IgE antibodies, and reduced numbers of nasal or bronchial mast cells, eosinophils, and CD4+ T-helper 2 (TH2) lymphocytes. Cytokine changes include reductions in serum interleukin (IL)-4 and in vitro lymphocyte-derived IL 4 as compared with a lack of increases in interferon gamma or IL-2. In a study in which skin biopsy specimens were obtained 24 hours after intradermal grass injection, the IT-treated patients had sharply increased numbers of macrophages which had messenger RNA for IL-12. This cytokine supports development of TH0 into TH1 CD4+ lymphocytes. Allergen immunotherapy results in a profound IgG and IgA antibody response with down-regulation of TH2 and possibly up-regulation of TH1 responses. PMID- 12528602 TI - Update on efficacy of allergen immunotherapy for allergic rhinitis and asthma. AB - Specific allergen immunotherapy (IT) has long been used to treat allergic rhinitis and asthma. Review of the literature from the past 5 years continues to support this use of IT as the only disease-modifying treatment of allergic disease currently available. In addition, studies suggest that allergen IT may prevent the progression of allergic disease. In monosensitized patients, IT may prevent polysensitization. In younger patients with allergic rhinitis only, IT may prevent the new onset of asthma. Although these studies require further validation, the evidence is important enough to consider disease prevention as one of the indications for prescribing allergen IT. PMID- 12528603 TI - Hymenoptera sensitivity: diagnosis and treatment. AB - Hymenoptera anaphylaxis is responsible for 50 deaths annually. This may be an underestimation because not all anaphylactic episodes are recognized or reported. Unexplained deaths at the poolside or on golf courses, as well as those attributed to cardiac causes, may be caused by unrecognized anaphylaxis. Venom immunotherapy (VIT) is highly effective in reducing an individual's risk of anaphylaxis. This review will discuss the diagnosis and treatment of hymenoptera sensitivity, patient selection for IT, standard VIT, and modified allergens. PMID- 12528604 TI - Novel immunologic therapies. AB - Although current allergen immunotherapy is an effective method of treating allergic inflammation, it is time consuming and carries the risk of anaphylaxis. Novel therapies for treating allergic inflammation are of two general types. In one senario, a key factor in the allergic cascade is blocked. The other possibility is to modify current immunotherapy administration. PMID- 12528605 TI - Comprehensive evaluation of cetirizine in the management of seasonal allergic rhinitis: impact on symptoms, quality of life, productivity, and activity impairment. AB - Improvement in patient daily functioning and well-being resulting from disease related symptom relief is increasingly viewed as a clinically relevant therapeutic outcome. The objective of this study was to evaluate the health related quality of life (HRQL) effects, safety and efficacy of cetirizine in the treatment of seasonal allergic rhinitis, and impact on self-reported work/school related productivity and activity impairment (WPAI). This was a double-blind, placebo-controlled, parallel-group study. Adult patients (n = 865) were randomized to daily treatment for 2 weeks with cetirizine or placebo. Patient disease-specific HRQL and WPAI were assessed at baseline and weeks 1 and 2 of treatment using the Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ) and Allergy-Specific Work Productivity and Activity Impairment (WPAI-AS) Instrument, respectively. Treatment with cetirizine resulted in greater (p < 0.001) improvement in overall RQLQ and individual domain scores (activities, sleep, non nose/eye symptoms, practical problems, nasal symptoms, eye symptoms, emotional), as compared with placebo. Cetirizine therapy produced a 28.9% mean reduction in total symptom severity complex (TSSC) score versus 12.7% with placebo at study end. Work/school productivity and activity impairment were significantly (p < 0.001) decreased from baseline for cetirizine-treated patients compared with placebo. The incidence of treatment-related side effects was similar between groups. Cetirizine provides safe and effective symptomatic relief in adults with SAR while significantly improving HRQL and providing a positive impact on work/school-related productivity and activity impairment. These results provide a more comprehensive assessment of cetirizine, indicating that its benefits extend beyond conventional measures of clinical outcome. PMID- 12528606 TI - Effect of respiratory syncytial virus infection during early infancy on the ontogeny of cytokine immune responses. AB - Recently, childhood asthma has been associated with an alarming rise in prevalence, morbidity, and mortality. Strong epidemiological evidence now links the development of childhood asthma with the persistence of T-helper lymphocyte type 2 (Th2) cytokine immune responses during early infancy. However, the ontogeny of human cytokine immune responses and the environmental factors influencing their development have not been fully elucidated. Several lines of evidence support the hypothesis that respiratory syncytial virus (RSV) infection during early infancy induces/amplifies the persistence of Th2 cytokine immune responses. Moreover, results from recent studies suggest that diminished interleukin (IL)-12 production by dendritic cells (DCs) is a potential mechanism by which RSV infection predisposes to the persistence of Th2 cytokine responses. Studies are needed to characterize the ontogeny of DC IL-12 production and Th cytokine responses during early infancy and to investigate the effect of RSV infection during early infancy on these responses. Results from such research may provide insight into the mechanism by which RSV infection may predispose to the development of childhood asthma. Moreover, they may result in the identification of potential strategies to attenuate or prevent the development of RSV infection and/or childhood asthma. Such strategies may include administration of IL-12 or manipulation of DC maturation or function. PMID- 12528607 TI - No growth suppression in children treated with the maximum recommended dose of fluticasone propionate aqueous nasal spray for one year. AB - This randomized, double-blind, placebo-controlled study of fluticasone propionate aqueous nasal spray (at a maximum recommended dose of 200 micrograms each day) administered daily for one year was conducted to evaluate its potential effects on growth, measured by stadiometry, in prepubescent children with perennial allergic rhinitis (n = 150; age 3.5 to 9.0 years). The results demonstrate equivalent growth velocity over a one-year treatment period between children receiving fluticasone propionate aqueous nasal spray and children receiving vehicle placebo. In addition, children in both treatment groups had similar increases in height over the same period. Baseline height was 119.1 cm (SE = 0.72) in the fluticasone propionate group (n = 56) and 119.0 cm (SE = 0.71) in the vehicle placebo group (n = 52). Mean height at the end of one year of treatment was 125.5 cm (SE = 0.18) in the fluticasone propionate group (n = 44) and 125.4 cm (SE = 0.19) in the vehicle placebo group (n = 39) (least-squares mean difference -0.12; 95% confidence interval [-0.600, 0.352]). The effects of fluticasone propionate on one-year growth velocity were also comparable to those of vehicle placebo. The confidence interval for the treatment difference lay within the prospectively defined equivalence bounds (of -0.8, +0.8 cm/year) and contained zero. The incidence of adverse events considered to be at least possibly drug-related did not differ between the fluticasone propionate group and the vehicle placebo group. The results of this one-year, double-blind study demonstrate that fluticasone propionate aqueous nasal spray at the maximum recommended dose of two sprays per nostril (200 micrograms) once daily was equivalent to vehicle placebo with no effects on growth rate in prepubescent children. PMID- 12528609 TI - HIV/AIDS surveillance and global estimates. Part II. PMID- 12528608 TI - Prolonged anticonvulsant hypersensitivity syndrome related to lamotrigine in a patient with human immunodeficiency virus. AB - Anticonvulsant hypersensitivity syndrome (AHS) is a rare, potentially life threatening allergic disorder, which is well described in relation to many aromatic anticonvulsants. Lamotrigine is a relatively new aromatic anticonvulsant agent that is thought to act on voltage-dependent sodium channels. Initially, it was licensed as add-on therapy for seizures inadequately controlled by other medications. However, its use has been broadened to other indications, including stand-alone therapy for seizures as well as for bipolar disorder. There is extensive experience with hypersensitivity syndromes related to phenytoin, carbomazepine, primidone, and phenobarbital, but fewer reactions have been reported to lamotrigine because of its relatively recent release. Patients with human immunodeficiency virus (HIV) have a higher rate of adverse reactions to many medications. It is unknown if they react more commonly to anticonvulsants such as lamotrigine. It is also unknown if the syndrome lias a tendency to be more severe or prolonged in such patients. The diagnosis of AHS may be particularly elusive in patients with HIV because its common features can easily be confused with an infectious etiology. We report the occurrence of a prolonged hypersensitivity syndrome likely related to lamotrigine in a 32-year-old female with HIV and review the literature regarding this condition. PMID- 12528610 TI - Meningitis stockpile for Africa. PMID- 12528611 TI - Protecting the rights of people with mental illness: can we achieve both good legal process and good clinical outcomes? PMID- 12528612 TI - [Pjotr Andreievich Kuprijanov (1893-1962)]. PMID- 12528613 TI - [Electrostimulation therapy--a new method of preventing postoperative wound complications in abdominal surgery]. AB - Experimental investigations have shown that chemotaxic activity of microorganisms in the parietal peritoneum and abdominal organ tissues is inhibited when they are treated with impulse electric discharges preventing the process of infecting the tissues. Results of the experiments underlay the development of a new method of prophylactics of postoperative complications in abdominal surgery based on the electroimpulsive treatment of the operation wound. Clinical approbation of the method showed its effectiveness, simplicity and safety. PMID- 12528614 TI - [Diagnosis and surgical treatment of pathologic tortuosity of carotid and vertebral arteries]. AB - The article summarizes an experience with 179 surgical interventions on patients with pathological tortuosity of brachiocephalic arteries. The optimal described diagnostic algorithm for this pathology included ultrasonic dopplerography, transcranial dopplerography, angiography or magnetic resonance tomography in the angiography regimen. Clinical and angiographic indications to surgical treatment of this pathology were determined. The operative treatment was carried on in patients with neurological symptoms having kinks of brachiocephalic arteries. The intraoperative strategy depending on the type and localization of the disease are analyzed. Stabile regression of manifestations of vascular cerebral insufficiency after surgical correction of the pathological tortuosity of the brachiocephalic arteries was achieved in 76% of the patients. PMID- 12528615 TI - [Clinico-anatomic rationale for the access and technique in video-endoscopic adrenalectomy]. AB - The results of surgical treatment of 60 patients with tumors of the adrenals with the help of endovideosurgical techniques were analyzed and described. The article also presents the data of experimental investigations performed in 31 corpses of subjects of different constitution and topography of the adrenals in the retroperitoneal space and different accesses to them with the help of endovideosurgical technique. The most effective and safe accesses and the method of endovideosurgical adrenalectomy have been developed, substantiated and approbated in clinical practical work. In most cases the right laparoscopic and left retroperitoneoscopic adrenalectomies should be considered the operations of choice in treatment of patients with tumors of the adrenals. It was proved that the decision for the optimal endovideosurgical access and the corresponding adrenalectomy should be made individually for each patient with special reference to the clinicoanatomical criteria found. The endovideosurgical adrenalectomy should not be opposed to open methods of surgery which are expedient in patients with large size of the tumor, signs of malignancy and possible intraoperative complications. PMID- 12528616 TI - [Preoperative embolization of the portal vein in malignant tumors of the liver: preliminary results]. AB - Preoperative embolization of the right portal vein and following hepatic resection were performed in seven patients with primary and secondary liver malignancies. The indication for the procedure was a small amount (30%) of the future remnant liver (FRL). The ultrasound-guided percutaneous transhepatic puncture of the portal vein and its embolization with gelatin sponge or silicon polymer was used. No complications of the procedure were noted. In 19-56 (mean 33) days computed or magnetic resonance tomography showed a significantly increased FRL volume (mean 40%). Right or extended right hepatectomy was performed in 5 and 2 patients respectively without lethal outcomes. A conclusion is made that the preoperative portal vein embolization is a safe and technically simple procedure that reduces the risk of postoperative hepatic failure after major liver resection. PMID- 12528617 TI - [Carcinoma of the major duodenal papilla]. AB - An experience with treatment of 62 patients with carcinoma of the major duodenal papilla during the period of 1989-1999 is generalized. Mean age of the patients was 62.4. Most of the patients admitted (85%) had jaundice. The most informative method of examination was FGDS, its sensitivity was 84%. All the patients were operated on. Transduodenal papillectomies (TDPE) were performed in 10 patients, pancreatoduodenal resection (PDR) in different modifications--in 42 patients, in 2 of them for recurrent carcinoma after TDPE, biliodigestive anastomoses--in 13 patients. Resectability was 80%, postoperative lethality--0%. Postoperative complications after PDR developed in 21% of the patients. Recurrent tumors after TDPE appeared in half of the patients. The actuarial five year survival after PDR was 50.6%. PDR is considered to be the operation of choice for cancer of the major duodenal papilla. PMID- 12528618 TI - [Characteristics of lymph flow in patients with post-mastectomy syndrome, based on lymphoscintigraphy data]. AB - Lymphoscintigraphy allows to get intravital contrast of functionally valuable lymph collectors of the extremity. A comparative analysis was made of the state of lymph flow in healthy subjects, in patients with breast cancer and in patients with the completed cycle of treatment of breast cancer having postmastectomy edema and without it. The investigation was performed in 280 extremities. Principal variants of the normal lymph flow (collector, diffuse, nodal) were marked out. Possible compensatory mechanisms ameliorating the draining function of the extremity were followed in patients with blocked lymph outflow. Programs of prognosis and treatment of patients with preclinical stage of postmastectomy edema are outlined. PMID- 12528619 TI - [Factors influencing the remote patency of distal shunts in patients with atherosclerosis of lower extremity arteries]. AB - Under study was the prognostic value of local risk factors of the development of late reocclusion of distal shunts in 149 patients with critical ischemia of lower extremities. It was found that the greatest influence on the remote patency of transplants is exerted by the state of the arterial wall in the area of the distal and in less degree of the proximal anastomoses. In a less degree the duration of functioning of the shunt depended on its length, position in soft tissues, transplant material and the level of the proximal anastomosis. The distal anastomosis level, the history of the simultaneous or previously made aorto-femoral shunt, geometry of the proximal and distal anastomoses did not have any reliable influence on the distal shunt patency. PMID- 12528620 TI - [False aneurysms of anastomoses after aorto-femoral reconstruction]. AB - The results of clinical, microbiological and histological studies in 141 patients with complications of the aorto-femoral graft revascularization were analyzed. The main group consisted of 44 patients with false aneurysms of vascular anastomoses. The findings obtained show that in 91.3% of cases the alterations in the wall of the artery adjacent to the anastomosis with false aneurysms resulted from chronic inflammatory reaction, the most common pathogens being St. Aureus and St. epidermidis. The optimal variants of revascularization of the extremity after resection are proposed. In case of distal anastomotic pseudoaneurysm the surgical strategy depends on the primary localization of the false aneurysm. PMID- 12528621 TI - [Active surgical management of critically burned patients (40 years of the method)]. AB - The system of active surgical management of the critically burned is based (besides the desire to begin the operative management as early as possible) on the complex of interrelated medical measures giving the greatest total gain of time having the decisive role for the successful recovery of the skin on the vast surface of the granulating wounds in the utmost concise terms before the development of irreversible changes in organism leading to death. The system was successfully used since 1960 in the Russian Nizhni Novgorod Burn Center and since 1973 in the Mordovian Republic Burn Center where 3000 patients with deep burns were treated. The system was used against the background of intensive infusion therapy during all the stages of burn disease and its complex treatment. When using this highly effective method, the possibility of recovery of these critically ill patients with deep burns previously thought to be doomed was shown. Results of the treatment were considerably better, the period of treatment at the hospital 2-3 times shorter and so the expenditure of the transfusion media and medicines became less. Lethality and invalidization of the recovered patients were much less which is of great social and economic importance. PMID- 12528622 TI - [A rare complication of conduction anesthesia in the treatment of finger injury]. AB - At the present time novocain still remains the anesthetic of choice in ambulatory surgery and traumatology. In practical work the 2% and 0.5% solutions are used. The 2% solution of novocain can in some cases (in patients prone to angiodystonic reactions) cause a persistent spasm of digital arteries that results in irreversible alterations in finger tissues down to the development of gangrene. This fact was confirmed in experiments with 2% solutions of novocain. It is proposed to follow the recommendations on using the 0.5% and 1% solutions of novocain for regional anesthesia in fingers. Such anesthetics as Lidocain and Trimecain should be used more widely since they possess more pronounced and continuous anesthetizing properties. PMID- 12528623 TI - [Extracorporeal methods of hemocorrection in the combined treatment of patients with purulent surgical infection of soft tissues complicated by sepsis]. AB - Extracorporeal methods of hemosorption and detoxication were used in treatment of 293 patients with purulent surgical infection of soft tissues complicated by sepsis. In 35 patients with the same nosology the methods in question were not employed. An analysis of the clinical observations has shown that lethality in the main group was reduced to 23.6%. The sorption technologies were shown to be most effective for normalization of leukocytosis, lekocytic index of intoxication, middle mass molecules in this group of patients. PMID- 12528624 TI - [Significance of invasive monitoring of central hemodynamics for the timely correction of their disturbances in operations on the abdominal aorta]. AB - Central hemodynamics was investigated with the thermistor Swan-Hanz catheter in 50 patients with the Leriche syndrome in whom aorto-femoral bifurcation shunting was performed. It was found that hemodynamic profiles of arterial hypotonia appearing at different stages of the operation can be expressed as hypovolemia, lower peripheral resistance of the vessels or the syndrome of low cardiac ejection. The therapeutic strategy (inotropic maintenance, vasodilators, infusion therapy) should be differentiated depending on the hemodynamic profile of hypotonia. PMID- 12528625 TI - [Weak stress factors in the prevention of postoperative complications in elective orthopedic surgery]. AB - Based on their personal and literature data the authors propose to use weak stressogenic factors (pyrogenal and interrupting normobaric hypoxia) as an element of preoperative preparation of patients for prevention of postoperative complications. The using of this method made the frequency of postoperative complications in the area of the operation wound twice as rare, outside the wound -three times as rare. PMID- 12528626 TI - [Method of laparoscopic catheterization of the cystic duct]. AB - During videolaparoscopic operations on bile eliminating pathways when the patent cystic duct is catheterized for cholangiography or drainage of choledochus by the Halsted-Pikovsky method there appear problems with the introduction of the tube into the cystic duct. In order to facilitate the cystic duct catheterization the authors propose to use a fishing-line conductor with the diameter of 0.8-1.0 mm. The method is simple, accessible, facilitates and accelerates the catheterization of the cystic duct and prevents compression of the tube lumen when fixing it with the endoclamp. PMID- 12528627 TI - [Surgical creation of a reservoir-colostomy with reflex evacuation]. AB - The authors consider that the problem of social rehabilitation of "colostomized" patients consists in an involuntary and sudden character of evacuation from the ostomy due to the absence of normal imperative urge to defecation. For this purpose a method of reservoir-retaining colostomy modulating the appearance of urge to defecation was worked out in experiments and used in clinic in 17 patients. Preconditions for the appearance of the urge were regular sensations due to mediated distention of the parietal peritoneum after filling the reservoir retaining volume. The subjective sensations preceding the evacuation from the colostomy appeared 10-20 minutes before defecation and were felt as discomfort and abdominal cramping towards of the external end of the colostomy in 15 (88.2%) and as elements of intestinal colic in 2 (11.8%) patients. So, 9 (53%) patients managed to sufficiently control the evacuation from the ostomy and not to use the pouch. Five patients (29.4%) demonstrated the ability to restrain defecation for 10-15 minutes by straining the anterior abdominal wall muscles when the urge appeared. Evacuation from the ostomy was reiterative (2-4 times), once or twice a day, rarely 3-4 times a day and lasted from 15 till 60 minutes. PMID- 12528628 TI - [Preliminary experience with partial ileoshunting in dyslipoproteinemia using endovideosurgical technique]. AB - The authors have proposed and fulfilled the operation of partial ileoshunting on a 45 year-old woman. The using of endovideosurgical method made the operation less traumatic and minimized the cosmetic defect. The patient was discharged on the 6th day. The authors have made 3 more operations of this kind in patients with hypercholesterinemia. PMID- 12528629 TI - [Treatment strategy in bleeding from gastroesophageal varicose veins in patients with liver cirrhosis complicated by portal hypertension]. AB - The article presents an analysis of results of different operations disuniting the gastroesophageal collector in patients with portal hypertension. Unsatisfactory long-term results of the Pazior operation widely used in CIS are known although this operation meets the requirements of emergency surgery aimed at saving the patient's life. The authors have developed an original method of disuniting the gastroesophageal collector. The method was tested in planned and emergency operations. An analysis of the preliminary results shows good perspectives of this direction. PMID- 12528630 TI - [Informative value of various methods of intraoperative biopsy of the pancreatic head]. AB - A comparative estimation of different methods of biopsy of the pancreatic head performed during operation on 192 patients with indurative diseases and cancer of the pancreatic head was made. The results of these investigations were compared with clinical and diagnostic findings before operation and with long-term results of the treatment. Two-lumen thick needle biopsy (the needle diameter 2 mm) of the pancreatic head proved to be most informative. In spite of the relatively high informative value of this method in cases of pseudotumorous pancreatitis (92%) the possibility of pancreatic head cancer is thought to be still underestimated in all diagnostic steps including the pre- and intraoperative stages. PMID- 12528631 TI - [Long-term results of pylorus-preserving gastric resections]. AB - An analysis of long-term results of pylorus-preserving resections of the stomach performed on 540 patients has shown that 44 patients (8.1%) had different postgastroresectional syndromes. Operations were made on 20 patients with severe manifestations of the postgastroresectional syndromes. An analysis of the causes of complications was made as well as the substantiation of the methods of reconstructive interventions. Indications and contraindications for pylorus preserving resections of the stomach are proposed. PMID- 12528632 TI - [Application of different textile vascular grafts in the aorto-femoral location and ways for their improvement]. AB - A comparative analysis of 220 reconstructions of the aortofemoral segment with the application of vascular prostheses of firms "Sever" and "Vascutek" was made. It was established that in patients with obvious anemia and hemodilution (hematocrit less than 25%, hemoglobin less that 80 g/l) an important condition for less total blood loss was to impart the zero surgical porosity to the graft wall at the preoperative stage. However, the biological impregnation used for these purposes reinforces the inflammatory component in the implantation of the vascular graft. The authors propose to use a more inert polytetrafluoroethylene film to impart the zero surgical porosity to the graft wall. PMID- 12528633 TI - [Successful treatment of a patient with a history of pancreatic necrosis and severe multi-organ insufficiency]. PMID- 12528634 TI - [A case of atypical course of carcinoid of the duodenum]. PMID- 12528635 TI - [Necrosis of the epiploic appendage in hernia of the line of Spigelius]. PMID- 12528636 TI - [Surgical sepsis. Part I. Immunologic markers of systemic inflammatory reaction]. AB - The aim of the work was to develop accessible methods of laboratory diagnosis of the systemic inflammatory reaction, based on an assessment of biological (inflammatory and immunosuppressive) activity of serum of surgical sepsis patients (n = 104). The investigation has shown that surgical sepsis patients are characterized by marked immune dysfunctions, immunosuppressive bioactivity of blood serum being detected in most patients (in 65.6%) beginning from the earliest stages of the development of the sepsis process. It was found that in patients with dominating suppressive activity of serum the immune disturbances were more pronounced and in every second case they were of combined type. The authors propose to use the degree of the immunosuppressive potential of serum factors as the key classification sign determining the presence of CARS-positive or CARS-negative phenotype in such patients. The results obtained suggest that it is expedient for complex therapy of surgical sepsis to include immunocorrection aimed at the weakening of immunosuppresive action of antiinflammatory mediators and shift of the balance towards the reinforcement of activity of proinflammatory ones (IL-12, IL-1) and Th-1 cytokines (IL-2, IFN-g). PMID- 12528637 TI - [Aleksei Alekseievich Troianov]. PMID- 12528638 TI - [The role of methyl methacrylate monomer in the development of bone cement implantation syndrome in endoprosthesis]. PMID- 12528639 TI - [Choice of plasma substitute in massive hemapheresis]. PMID- 12528640 TI - PDAs are willing, but will they be able? PMID- 12528641 TI - Speech recognition technology gives voice to clinical data. PMID- 12528642 TI - E-health demand may finally catch up with supply. PMID- 12528643 TI - Biometrics: can we have a show of hands? PMID- 12528644 TI - Technological advances could change the face of telemedicine. PMID- 12528645 TI - [Impact of intravenous thrombolysis prior to percutaneous coronary intervention in reperfusion therapy for acute myocardial infarction]. AB - OBJECTIVES: To elucidate the effectiveness and safety of intravenous thrombolysis (IVT) with mutant tissue plasminogen activator prior to percutaneous coronary intervention (PCI) in patients with acute myocardial infarction. METHODS: Ninety consecutive patients were recruited with the following criteria: acute myocardial infarction with ST segment elevation or bundle branch block on electrocardiography, admission within 6 hr from onset, age of < or = 80 years and without previous PCI or coronary bypass graft surgery. They were divided into two groups. Group IV consisted of 53 patients treated with IVT prior to PCI and Group D consisted of the other 37 patients with direct PCI. Mutant tissue plasminogen activator, monteplase, was administered with a dose of 27,500 U/kg in Group IV (maximum injection dose, 160 x 10(4) U). The clinical features and in-hospital outcome were compared between the two groups. RESULTS: Patients in Group IV acquired earlier reperfusion estimated by electrocardiography recovery at 60 min after admission and higher Thrombolysis in Myocardial Infaction (TIMI) flow grade on the first coronary angiogram (TIMI 2 or 3 flow rate; Group IV vs Group D = 75% vs 35%, p < 0.0001). The duration from onset to TIMI 3 flow grade was not significantly different between Group IV and Group D (230 vs 260 min, p = 0.15). The incident of ST segment re-elevation with chest pain at recanalization was lower in Group IV than in Group D (23% vs 46%, p < 0.05). The duration from TIMI 3 recognition to peak creatine kinase level was longer in Group IV (466 vs 359 min, p = 0.039). Subacute thrombotic occlusion occurred in two patients in Group IV and three in Group D (NS). One patient in each group died from pump failure (NS). No severe bleeding complication was found in any patient. CONCLUSIONS: IVT prior to PCI was considered to be a safe, effective and useful therapy in patients with acute myocardial infarction. Different patterns of reperfusion might occur, because of the low frequency of ST re-elevation and elongation of duration from reperfusion to peak creatine kinase level in patients treated with IVT prior to PCI. PMID- 12528646 TI - [Measurement of coronary flow reserve by pressure/temperature sensor guide wire based thermodilution in experimental models]. AB - OBJECTIVES: Recently, a combined 0.014 pressure/temperature sensor-mounted guide wire has been developed to simultaneously measure fractional flow reserve and coronary flow reserve (CFR) by thermodilution (CFR-thermo). The accuracy of CFR thermo was compared with CFR obtained by flow rate (CFR-flow) in experimental models. METHODS: Using an experimental model made from a straight-rigid tube (4 mm diameter) filled with 36 degrees C water, CFR-thermo and CFR-flow were measured under different conditions of sensor position and injected water temperature (0-40 degrees C). A side branch (2 mm diameter) was then placed at 4, 6, 8 and 10 cm from the injected site just proximal to the stenosis. The degree of stenosis ranged from 0 to 75% (0%, 25%, 50%, 75%). CFR-thermo and CFR-flow were calculated from the inverse ratio of the mean transit time and the flow ratio during high flow to low flow rates. RESULTS: Under the conditions without the side branch, there were good correlations between CFR-thermo and CFR-flow if the temperature of the injected water was under 28 degrees C and the sensor was not placed within 4 cm from the injection site. With the side branch, CFR-thermo was smaller than CFR-flow although there were good correlations between CFR thermo and CFR-flow. The value of CFR-thermo increased with more distal positions of the side branch to the injected site. CONCLUSIONS: Temperature of the injected water, and the position of the sensor, the side branch and the stenotic lesion may influence measurements of CFR-thermo. These effects should be considered when CFR is measured by the thermodilution method. PMID- 12528647 TI - [Usefulness of left ventricular opacification with intravenous contrast echocardiography in patients with asymptomatic negative T waves on electrocardiography]. AB - OBJECTIVES: Patients with electrocardiography (ECG) abnormalities sometimes present without obvious symptoms or abnormal physical findings. In some cases, echocardiography fails to reveal abnormalities compatible with the ECG findings because of poor echocardiographic image quality. New intravenous contrast agents now enable opacification of the left ventricular cavity in the clinical setting. The usefulness of left ventricular opacification (LVO) by ultrasonic contrast agent was examined in asymptomatic patients with negative T waves on ECG. METHODS: Thirty-four consecutive patients (23 males, 11 females, mean age 68 +/- 10 years) with negative T waves without symptoms or abnormal physical findings underwent routine transthoracic echocardiography and LVO by intravenous injection of Levovist using harmonic imaging. RESULTS: Without LVO, abnormal echocardiographic findings compatible with the negative T waves were identified in 22 (65%) of the 34 patients. With LVO, the endocardial border of the left ventricle was observed in all patients and findings compatible with negative T waves were detected in 32 (94%) of the 34 patients, which was significantly higher than that without LVO (chi 2 = 9.79, p = 0.0055). Furthermore, LVO revealed additional findings in nine patients such as mural thrombus or apical thinning. In total, LVO revealed abnormal apical findings in 22 patients (65%), which was significantly higher than that without LVO (chi 2 = 10.0, p = 0.0013). CONCLUSIONS: LVO is useful for identifying abnormal findings, especially in the apex in asymptomatic patients with negative T waves on ECG. PMID- 12528648 TI - [Clinical experience with retrievable vena cava filters for prevention of pulmonary thromboembolism]. AB - OBJECTIVES: To evaluate the feasibility, effectiveness and complications of the retrievable vena cava filter [Guther tulip vena cava filter(GTF)] for the prevention of pulmonary thromboembolism in patients with deep vein thrombosis. METHODS: Seventeen patients, 3 males and 14 females, aged 21 to 82 years (mean age 59 +/- 19 years), underwent implantation of GTFs between December 2000 and February 2002 at Mie University Hospital. All patients were treated under diagnoses of deep vein thrombosis with or without pulmonary thromboembolism based on venous ultrasonography, venography or computed tomography. Eleven patients were treated with thrombolysis. RESULTS: Significant thromboembolus was trapped within the filter in 3 of 12 patients. No acute pulmonary thromboembolism occurred during implantation or at retrieval of the GTF. Retrieval of the GTF was attempted in 9 patients, and 8 GTFs were retrieved successfully. Mean interval of the filter implantation was 13.4 +/- 6.3 days and the mean retrieval time was 4.8 +/- 3.2 min. No complications occurred except for one case of minor hemorrhage at the puncture site. CONCLUSIONS: The placement and retrieval of the retrievable vena cava filter was feasible and safe. This filter was also effective for the prevention of pulmonary thromboembolism. This retrievable vena cava filter may be a good first-choice filter for both permanent and temporary use. PMID- 12528649 TI - Idiopathic left ventricular aneurysm associated with pleomorphic ventricular tachycardia: a case report. AB - A 39-year-old Japanese woman presented with an idiopathic left ventricular aneurysm manifesting as recurrent episodes of palpitation. She was referred to our hospital for evaluation of sustained ventricular tachycardia. Echocardiography disclosed a dyskinetic well-defined wall bulge during both systole and diastole at the basal region of the interventricular septum, and reduced left ventricular wall thickness and severe hypokinesis at the anterolateral to posterolateral region. These appearances were confirmed by the angiographic findings. The sustained ventricular tachycardia was reproducibly induced by a single extrastimulus from the right ventricular apex. Subsequently, 4-type ventricular tachycardias were induced during the electrophysiological study and the mechanism of these ventricular tachycardias was considered reentry. Radiofrequency catheter ablation failed due to the changing QRS morphologies during the entrainment study. The patient was treated with cibenzoline 300 mg a day, and there has been no recurrence of tachycardia during the 18-month follow up period. PMID- 12528650 TI - [Follow-up coronary angiography with multi-slice computed tomography after stenting]. PMID- 12528651 TI - Medicine in academia in the 21st century. PMID- 12528652 TI - Differentiating managed utilization vs. care. PMID- 12528654 TI - Between medicine, technology, industry, & public health. PMID- 12528653 TI - Mobile radiography CQI: an inter-national study. AB - Mobile or bedside radiography has been and is a staple diagnostic and follow-up tool used readily by the many medical disciplines, such as cardiology, surgery, orthopedics, pediatrics, neonatology, etc. Ironically, in the past a student or the least qualified technologist was sent to perform the bedside exam. Moreover, it was almost expected that poor but acceptable film quality would result or that repeat films were almost always to be taken. Inefficiency with respect to quality of exam, the time the exam takes, or film repeats can be costly. The price of inefficiency is the cost involved in doing things incorrectly or not in the most efficient manner, i.e., incurring inefficiencies instead of operating in an ideal manner. The purpose of this study was to compare the total cost of inefficiently organized, scheduled and performed mobile radiography at three large teaching hospitals in various locations and of diverse patient loads, as a means of determining how best to increase utilization and performance. The study was performed at the 489-bed New England Deaconess Hospital (NEDH), the 644-bed Sentara Norfolk General Hospital (SNGH), and the 1500-bed Rheinische Westfalische Technische Hochschule (RWTH) in Aachen, Germany. Similar standardized study methods were utilized at all three institutions where extended observation of mobile utilization, areas of inefficiency, time wasted per episode and number of episodes per time period were determined. Data were loggedin at three standardized time periods, summated, and then multiplied by technologist hourly pay rate. This sum was extrapolated over 52 weeks to give the total annual cost of inefficiently organized mobile radiography. For NEDH the cost of total inefficiency was $75,453, for SNGH $49,586, while for RWTH it was $9,519. Eighteen areas of inefficiency were identified and grouped, such as lack of spatial cohesiveness and lack of communication leading to film duplication, etc. While inefficiencies in the delivery of hospital based health care are well known, this study attempts to quantify and determine a dollar value for each process found as inefficient. Key inefficiencies were found to be common at large hospitals no matter whether in the United States or Europe. These impairments are responsible for a disproportionate share of overall inefficiency, and their elimination (achievable by simple solutions) would result in drastic cost reductions (ranging from 40-75% at the institutions studied). Thus this study is important in view of spiralling costs, as it is a key component of total quality management (TQM) in radiology and a continuous quality improvement (CQI) tool for mobile radiology specifically. PMID- 12528655 TI - Osteoporosis: boning-up on the essentials. PMID- 12528656 TI - The effects of Medicare payment subsidies to teaching hospitals. AB - The Medicare program is the nation's largest single source of funds for graduate medical education. The program pays teaching hospitals for the direct costs of their residency programs and, since 1983, has paid for some of the indirect costs of graduate medical education. The rationale for, and extent of, the payments for indirect costs have been debated for years; recently, Congress has reduced the payments as it attempts to rein in Medicare costs. This Issue Brief reviews the status and recent history behind indirect medical education (IME) payments, and summarizes research that investigates how hospitals have responded to the incentives created by these and other supplemental payments. PMID- 12528657 TI - Effect of UV irradiation on organic matter extracted from treated Ohio River water studied through the use of electrospray mass spectrometry. AB - Ohio River water was treated by settling, sand filtration, and granular activated carbon filtration. It was then irradiated by low-pressure (monochromatic) and medium-pressure (polychromatic) UV lamps to investigate the effects of UV irradiation on the extracted organic matter (EOM). When the EOM, collected by solid phase extraction cartridges, was analyzed by conventional UV spectroscopy and size exclusion chromatography (SEC), no significant changes in the EOM were revealed for various UV doses. Positive and negative electrospray ionization mass spectrometry (ESI-MS) of the EOM produced mass spectra that vary significantly with UV dose. The UV dosage conditions also appear to affect the reactivity of the EOM to subsequent chlorination. The magnitude of the spectral changes is generally greater for medium-pressure lamps than for low pressure and increases with UV exposure. Based on the observed MS peaks, the changes may be due to the presence of lignin, resulting perhaps from photooxidation and/or photo rearrangement of macromolecules in the sample. When chlorination is used for secondary disinfection, these results suggest that it may be important to consider the effects of UV irradiation on the organic matter in the water before applying UV disinfection technology to a particular source water. PMID- 12528659 TI - Recession and anterior transposition of the inferior oblique muscle [RATIO] to treat three cases of absent inferior rectus muscle. AB - BACKGROUND AND PURPOSE: The "loss" of an extraocular muscle during a surgical procedure requires a prompt solution. Muscle agenesis produces a strabismus of difficult resolution. In these cases, alternative strategies are required to restore the binocular alignment. We present three consecutive cases of inferior rectus absence that were successfully surgically resolved. SUBJECTS AND METHODS: Three children with absent inferior rectus muscles are presented; one due to loss during strabismus surgery and two due to inferior rectus agenesia. All were treated with recession and anterior transposition of the inferior oblique muscle [RATIO]. RESULTS: All patients achieved a satisfactory result with the technique employed. CONCLUSION: RATIO is a useful procedure to restore binocular alignment in cases of inferior rectus absence. PMID- 12528660 TI - A reply to a previous editorial on the International Strabismus Association. PMID- 12528662 TI - [Comments on "Herman Simon"]. PMID- 12528664 TI - [Author reply]. PMID- 12528663 TI - [Comments on the article, "Mental disability and incompetence to make decisions and correlation with legal guardianship"]. PMID- 12528665 TI - Space life sciences: biological research and space radiation. Proceedings of the F1.2, F1.3, F2.2 and F2.6 Symposia of COSPAR Scientific Commission F which were held during the Thirty-third COSPAR Scientific Assembly, Warsaw, Poland, July, 2000. AB - This issue of Advances in Space Research contains a large number of manuscripts in the discipline of Space Life Sciences including papers from the following sessions of the Warsaw COSPAR Assembly: Gravity-related research with animals- past, present, future; The nervous system: space flight environmental factors effects--present results and new perspectives; Investigating space radiation effects at particle accelerators--biology and physics experiments; Perspectives on radiation risks on long space missions: deterministic and stochastic effects. PMID- 12528666 TI - Gravity related research with fishes--perspectives in regard to the upcoming International Space Station, ISS. AB - During the entire evolution of life on Earth, the development of all organisms took place under constant gravity conditions, against which they achieved specific countermeasures for compensation and adaptation. On this background, it is still an open question to which extent altered gravity such as hypergravity (centrifuge) or microgravity (spaceflight) affects the normal individual development, either on the systemic level of the whole organism or on the level of individual organs or even single cells. The present review provides information on these questions, comprising gravistimulated effects on invertebrates and vertebrates (with the exception of mammals, since respective biomedically oriented reviews abound), focusing on developing fish as model systems, with special emphasis on the effect of altered gravity on the developing brain and vestibular system, comprising investigations on behaviour and plastic reactivities of the brain and inner ear. Clues and insights into the possible basic causes of space motion sickness-phenomena (SMS; a kinetosis) are provided as well as perspectives in regard to future work to be done including studies on the ISS concerning the analysis of gravistimulated effects on developmental issues (imprinting phase for graviperception?). PMID- 12528667 TI - Directional filling-in. AB - The filling-in theory of brightness perception has gained much attention recently owing to the success of vision models. However, the theory and its instantiations have suffered from incorrectly dealing with transitive brightness relations. This paper describes an advance in the filling-in theory that overcomes the problem. The advance is incorporated into the BCS/FCS neural network model, which allows it, for the first time, to account for all of Arend's test stimuli for assessing brightness perception models. The theory also suggests a new teleology for parallel ON- and OFF-channels. PMID- 12528668 TI - Adaptation of the macular vestibuloocular reflex to altered gravitational conditions in a fish (Oreochromis mossambicus). AB - Young fish (Oreochromis mossambicus) were exposed to microgravity (micro g) for 9 to 10 days, or to hypergravity (hg) for 9 days. For several weeks after termination of micro g and hg, the roll-induced static vestibuloocular reflex (rVOR) was recorded. In stage 11/12-fish, the rVOR amplitude (angle between the maximal up and down movement of an eye during a complete 360 degree lateral roll) of micro g-animals increased significantly by 25% compared to 1 g-controls during the first post-flight week but decreased to the control level during the second post-flight week. Microgravity had no effect in stage 14/16 fish on the rVOR amplitude. After 3 g-exposure, the rVOR amplitude was significantly reduced for both groups compared to their 1 g-controls. Readaptation to 1 g-condition was completed during the second post-3 g week. We postulate a critical period during which the development of the macular vestibuloocular reflex depends on gravitational input, and which is limited by the first appearance of the rVOR. At this period of early development, exposure to microgravity sensitizes the vestibular system while hypergravity desensitizes it. PMID- 12528669 TI - [The Internet and its security]. AB - Internet, is the greatest world net by by means of which nowadays the planet communicates, rapidly goes forward. The last years of the university in USA the commonly develop the more progressive concept of the net (Internet 2), thanks to the constant growing technologies, with the goal to answer the needs of the scientific and the educational institutions, but also the commercial institutions and the organizations. Almost the there is no more significant institution in the world which has not developed their web pages and data bases with the most actual contents available to the wider circle of the users. In this paper we have given the section of the most actual web pages. However, Internet is not immune to those users who are not benevolent and who have developed the different tools in the goal of the destroying or unabling of the normal use of all the Internet conveniences. The authors is considering the protection problem and the data security which get distributed by Internet. PMID- 12528670 TI - Influence of hypergravity on fish inner ear otoliths: I. Developmental growth profile. AB - Inner ear stones (otoliths) of larval cichlid fish Oreochromis mossambicus were marked with the calcium-tracer alizarin-complexone (AC) at 1 g earth gravity before and after a 3, 7, 14 or 21 days stay of the animals at hypergravity conditions (hg; 3 g, centrifuge). After the experiment, the otoliths' area between the two AC-labellings was measured with regard to size and asymmetry (size difference between the left and the right stones). Both utricular and saccular otoliths (lapilli and sagittae, respectively) continued growing in a linear way at hg, but growth was significantly slowed down as compared to parallely raised 1 g-control specimens. In case of bilateral asymmetry between the corresponding otoliths its formation in hg-animals became reduced as compared to the 1 g controls. The reduction of asymmetry was much more pronounced in the sagittae than in the lapilli. The latter result supports an earlier hypothesis, according to which especially a low sagittal asymmetry has a functional advantage. In general, the results strongly suggest that otolith growth is continuously regulated in dependence of the environmental gravity vector. PMID- 12528671 TI - [Advanced Trauma Life Support (ATLS)]. PMID- 12528672 TI - Influence of hypergravity on fish inner ear otoliths: II. Incorporation of calcium and kinetotic behaviour. AB - Larval siblings of cichlid fish (Oreochromis mossambicus) were subjected to hypergravity (hg; 3 g, 14 days) during development. Following the transfer to 1 g (i.e., stopping the centrifuge) they were separated into normally and kinetotically swimming individuals (the latter performed spinning movements). During hg, the animals were maintained in aquarium water containing alizarin complexone (AC), a fluorescent calcium tracer. Densitometric measurements of AC uptake into inner ear otoliths (optical density of AC/micrometers2) revealed that the kinetotic individuals had incorporated significantly more AC/calcium than the normally behaving fish. Since the amount of otolithic calcium can be taken as an approximation for otolith weight, the present results indicate that the otoliths of kinetotically swimming samples were heavier than those of the normally behaving larvae, thus exhibiting a higher absolute weight asymmetry of the otoliths between the right vs. the left side of the body. This supports an earlier concept according to which otolith (or statolith) asymmetry is the cause for kinetoses such as human static space sickness. PMID- 12528673 TI - Swimming behavior of larval Medaka fish under microgravity. AB - Fish exhibit looping and rolling behaviors when subjected to short periods of microgravity during parabolic flight. Strain-differences in the behavioral response of adult Medaka fish (Oryzias latipes) were reported previously, however, there have been few studies of larval fish behavior under microgravity. In the present study, we investigated whether microgravity affects the swimming behavior of larvae at various ages (0 to 20 days after hatching), using different strains: HNI-II, HO5, ha strain, and variety of different strains (variety). The preliminary experiments were done in the ground laboratory: the development of eyesight was examined using optokinetic response for the different strains. The visual acuity of larvae improved drastically during 20 days after hatching. Strain differences of response were noted for the development of their visual acuity. In microgravity, the results were significantly different from those of adult Medaka. The larval fish appeared to maintain their orientation, except that a few of them exhibited looping and rolling behavior. Further, most larvae swam normally with their backs turning toward the light source (dorsal light response, DLR), and the rest of them stayed with their abdomen touching the surface of the container (ventral substrate response, VSR). For larval stages, strain differences and age-differences in behavior were observed, but less pronounced than with adult fish under microgravity. Our observations suggest that adaptability of larval fish to the gravitational change and the mechanism of their postural control in microgravity are more variable than in adult fish. PMID- 12528674 TI - Interobserver variability of magnetic resonance angiography. PMID- 12528675 TI - Hits and misses in nicotine psychopharmacology: a personal view of research over a period of 30 years. PMID- 12528676 TI - On autonomy and participation in rehabilitation. AB - PURPOSE: To explore the concept of autonomy as a basis for social participation, with particular reference to rehabilitation. METHOD: A study of relevant literature from the field of rehabilitation, building on theory developed in other fields (ethics, social sciences), and deriving important concepts and strategies for rehabilitation practice. RESULTS: The focus of rehabilitation for people with a chronic disabling condition is shifting from a biomedical to a client-centred perspective. Conceptions of autonomy vary among individuals and cultures, but a crucial distinction can be made between decisional autonomy (the ability to make decisions without external restraint) and executional autonomy (the ability to act as one wishes). The liberal-individualist account of autonomy over-emphasizes physical independence and does not sufficiently recognize the inter-dependency of all people, including those with disabilities. An ethic of care, complementary to the principle of respect for autonomy, should guide the development of rehabilitation strategies to enhance individual autonomy and participation in daily living. For rehabilitation, this entails an attentive attitude, maximizing opportunities for informed choices, taking full account of each person's preferences, needs and social contexts. CONCLUSIONS: Autonomy is central to client-centred rehabilitation since it is a pre-requisite for effective participation. It is suggested that autonomy, conceived as a basis for participation, is the ultimate aim of rehabilitation. PMID- 12528685 TI - PGD: new ethical challenges. PMID- 12528687 TI - Persistent vegetative state: let's not blow out the candle. PMID- 12528689 TI - Reactive astrocytes within the acute plaques of multiple sclerosis are PSA-NCAM positive. PMID- 12528690 TI - Temozolomide and cisplatin in the treatment of leptomeningeal metastatic involvement from melanoma: a case report. PMID- 12528691 TI - Project Cronos: multiple versions of the Mini Mental State Examination. PMID- 12528692 TI - Staging of the intracerebral inclusion body pathology associated with idiopathic Parkinson's disease (preclinical and clinical stages). AB - The synucleinopathy known as idiopathic Parkinson's disease (IPD) is a multi system disorder in the course of which only a few predisposed nerve cell types in specific regions of the human brain become progressively involved. The underlying neuropathological process (formation of proteinaceous intraneuronal inclusion bodies) intracerebrally begins in clearly defined induction sites and advances in a topographically predictable sequence. Components of the autonomic, limbic, and motor systems sustain especially heavy damage. During the presymptomatic stages 1 and 2, the IPD-related inclusion body pathology remains confined to the medulla oblongata and olfactory bulb. In stages 3 and 4, the substantia nigra and other nuclear grays of the midbrain and basal forebrain are the focus of initially subtle and, then, severe changes. The illness reaches its symptomatic phase. In end-stages 5 and 6, the pathological process encroaches upon the telencephalic cortex. IPD manifests itself in all of its dimensions, which under the influence of the supervening cortical pathology are subject to increasing complexity. PMID- 12528693 TI - On dying well: how does one live spiritually in the hope of dying well? PMID- 12528694 TI - Patient no more: why did the golden age of medicine collapse? PMID- 12528695 TI - Congenital arthrogryposis in pregnancy. PMID- 12528696 TI - Counting the cost: the use of financial incentives in managed care. PMID- 12528697 TI - Clinical features and laboratory findings for differentiating parkinsonian syndromes. AB - Most clinicians and researchers still accept idiopathic Parkinson's disease as a disease entity, although the identification of several genes or loci responsible for certain inherited forms of Parkinson's disease have provided arguments for a clinical heterogeneity of this disorder. The here proposed new classification of syndromes and diseases which show a parkinsonian syndrome in isolation or in combination with other symptoms divides patients into groups based on a set of rules mainly on a clinicopathological basis. It illustrates the current state of the art and will eventually be replaced due to an increase in the knowledge of the pathophysiological basis of these diseases. According to this classification, the crucial clinical features and laboratory findings are reviewed. The report will mainly concentrate on parkinsonian syndromes in degenerative diseases. PMID- 12528698 TI - Heroin overdose in pregnancy: an unusual case report. PMID- 12528699 TI - A curriculum for teaching clinical ethics in neonatal-perinatal medicine. AB - Ethical thought plays a role in clinical decision-making, especially in fields such as neonatal-perinatal medicine. There is limited information, however, on the best way to teach ethics to postgraduate medical trainees in this subspeciality. This article describes the development, implementation, and evolution of a structured curriculum in ethics for a program in neonatal perinatal medicine. The curriculum is facilitated by a specialist in ethics and based on hypothetical case scenarios, which are discussed using a framework that highlights the ethical values promoted or neglected in each decision option. Formal evaluation of the program has been impossible, but informal reviews indicate that there is a high degree of satisfaction. PMID- 12528701 TI - Multiple pregnancy: delayed delivery and outcome. PMID- 12528700 TI - Colonic pseudo-obstruction following elective caesarean section. PMID- 12528702 TI - Congenital heart block presenting as persistent fetal bradycardia. PMID- 12528703 TI - Difference or defect? Intersexuality and the politics of difference. AB - Under current medical practice, when a child is born with ambiguous genitalia, sex is assigned and medical/surgical intervention undertaken. This practice is criticized by the Intersex Society of North America and by feminist scholars. Together they are creating a "politics of difference" approach in which differences are not seen as "defects" to be corrected. This paper analyzes the reasons offered in support of current medical practice and of the politics of difference, and argues for a move toward the latter. Crucial to the determination that this move is warranted are justice concerns and the emergence of support groups and technologies that allow geographically separated individuals to form a relevant "group." PMID- 12528704 TI - Relationship between head orientation and torsional eye movements in goldfish during linear acceleration. AB - We analyzed torsional eye movements of normal goldfish during sinusoidal linear acceleration, altering the orientation of the fish on the linear accelerator in the yaw plane over a range of 90 degrees and in the pitch plane up to 30 degrees. We video-recorded changes of torsional eye movements associated with a body rotation in the yaw and pitch plane and analyzed them frame by frame. In normal fish, we observed clear torsional eye movements for stimuli of 0.1 G linear accelerations along the body axis in the horizontal position. Torsion occurred in the opposite direction of resultant force produced by linear acceleration and gravity. Though the amplitude of these compensatory responses increased with increasing magnitude of acceleration up to 0.5 G, the torsion angle did not fully compensate the angle calculated from gravity and linear acceleration. Furthermore, the torsion angle decreased as the longitudinal body axis deviated from the direction of linear acceleration. For the body axis perpendicular to the direction of acceleration, torsional eye movement was still observed. When we tilted the fish in the pitch plane, compensatory eye torsion occurred. The response amplitude to acceleration decreased for both head-up and head-down up to 30 degrees. These results suggested the existence of specific connections between the otolith organ and ocular muscles. PMID- 12528705 TI - Making health care decisions without a prognosis: life in a brain trauma unit. AB - When the author's daughter was hit by a car and remained unconscious for seven months, she found that there were certain factors where traditional ethical theory was not sufficiently nuanced to guide her practical decision making in regard to her daughter's health care. This article concentrates on three of those factors. They are: (1) no reliable prognosis can be offered for many brain injured individuals; (2) a patient's age and the relationship between the patient and the caregiver affect the context of caring; and (3) there are severe difficulties in obtaining and sustaining chronic care and accessing scarce resources. PMID- 12528706 TI - The forgotten IUCD--a potential problem for post-menopausal women. PMID- 12528707 TI - A cabbit in sheep's clothing: exploring the sources of our moral disquiet about cloning. AB - Emerging from the first successful cloning of a mammal, a sheep named "Dolly," is a critical but under-asked question: "Why do so many of us find this feat (and its potential application to human subjects) to be deeply disturbing?" This paper suggests that the answer rests not primarily in the theological and philosophical arguments most often heard against cloning but in the threat the act poses to our foundational "cosmological categories." Building upon theories introduced by Mary Douglas and Jeffrey Stout, the essay argues that Dolly becomes a "cabbit" in sheep's clothing--an outwardly innocuous entity which, like Stout's cat/rabbit, offends at a deep and visceral level. Like the cabbit, the cloned sheep and especially the cloned human are disturbing not because of the way they are produced nor by the physical threat they pose but because they challenge the very way we understand and organize our world. PMID- 12528708 TI - Otoliths as biomechanical gravisensors. AB - This paper analyzes experimental data related to the reaction of otolith afferents in response to acceleration (Fernandez and Goldberg, 1976). It considers the assumptions that were the basis of the interpretation of the stimulus-response characteristics of afferents proposed by Fernandez and Goldberg. Comparing these experimental data with the results of modeling the otolith structures of vertebrates indicates that some peculiarities of the neural responses may be explained by the spatial dependence of the material parameters of the otolithic membrane across its thickness and within the volume of the membrane corresponding to the terminal field. The importance of the spatial dependence of the material parameters of the otolithic membrane for otolith functioning is discussed. PMID- 12528709 TI - Rapid incarceration of a ring pessary with its safe and immediate removal. PMID- 12528710 TI - Ovarian function during chemotherapy with paclitaxel and carboplatin. PMID- 12528711 TI - Introduction: doing health care ethics today. PMID- 12528712 TI - Secondary amenorrhoea due to the use of rifampicin as an antimycobacterial agent. PMID- 12528713 TI - Persistent IGF-I overexpression in skeletal muscle transiently enhances DNA accretion and growth. AB - Adult transgenic mice with muscle-specific overexpression of insulin-like growth factor (IGF)-I have enlarged skeletal muscles. In this study, we; 1) characterized the development of muscle hypertrophy with respect to fiber type, age, and sex; 2) determined the primary anabolic process responsible for development of hypertrophy; and 3) identified secondary effects of muscle hypertrophy on body composition. Transgene expression increased with age and was present only in fibers expressing type IIB fast myosin heavy chain. Muscle masses were greater by 5 wk of age, and by 10 wk of age the differences were maximal despite continued transgene expression. Total DNA and RNA contents of the gastrocnemius muscle were greater for transgenic mice than for nontransgenic littermates. The differences were maximal by 5 wk of age and preceded the increase in protein mass. The accelerated protein deposition ceased when the protein/DNA ratio attained the same value as in nontransgenic controls. Despite localization of IGF-I expression to muscle without changes in plasma IGF-I concentrations, genotype also modified the normal age and sex effects on fat deposition and organ growth. Thus, enhanced DNA accretion by IGF-I was primarily responsible for stimulating muscle growth. In turn, secondary effects on body composition were incurred that likely reflect the impact of muscle mass on whole body metabolism. PMID- 12528714 TI - Methodology and theology in health care ethics. PMID- 12528715 TI - Medical prevention of space motion sickness--animal model of therapeutic effect of a new medicine on motion sickness. AB - Space motion sickness (MS) is one of the most important problems in the field of space medicine. In order to prevent space MS, a new medicine, PMPA, has been prepared by means of synthesizing in our laboratory. The purposes of this study were to set up animal models of PMPA against MS, and to observe its effects on anti-MS, and to prove its function of antagonism to choline. Eight cats, forty rabbits and two hundred and ten rats were selected as animal subjects. The parallel swing stimulus, a method causing the reversal syndromes and tests of anti-choline function were used in our experiments. The results are as follows: (1) The score of MS symptoms in cats with PMPA or scopolamine (SCOP) is significantly lower than that in cats with placebo (p<0.01), while the incidences of efficiency and prevention of PMPA (87.5%, 75%) are higher than those of SCOP (75.0%, 50%) in cats. (2) PMPA of 1.6 mg/kg or 0.8 mg/kg could antagonize the reversal syndromes and repress reversal rotation significantly in rabbits like SCOP in comparison with placebo (p<0.01). (3) PMPA could inhibit tremor evoked by oxotremorine or by nicotine-procaine in rats like SCOP, and play an important role in the antagonism to central M-choline and N-choline receptors. The animal experiments demonstrate that PMPA is an effective medicine against MS with antagonism function to choline. PMID- 12528716 TI - Ethics and human genetics. PMID- 12528717 TI - Pregnancy in Takayasu's arteritis with thrombophilia. PMID- 12528718 TI - The ethics of access to health care. PMID- 12528719 TI - Ethical issues at the end of life. PMID- 12528720 TI - Hypovolaemic shock secondary to epistaxis in pregnancy. PMID- 12528721 TI - Patient-provider relations--understanding the social and cultural circumstances of difficult patients. AB - Clinicians who have understanding and insight into the social and cultural background of their patients will be better prepared to foster the mutual respect required for effective chronic disease management. This paper will develop an argument linking respect in patient-physician relations to the social determinants of health. PMID- 12528722 TI - Urodelean amphibians in studies on microgravity: effects upon organ and tissue regeneration. AB - Results obtained from nine experiments performed onboard Russian biosatellites have shown that microgravity promotes tissue regeneration in the newt, Pleurodeles waltl. The effect has been reproduced in all flights and on a clinostat as well for eye tissues (lens and retina), limbs and tail. The effect was demonstrated in 1.5- to 2-fold increase in cell proliferation in the early stages of regeneration in space flight. Animals "flown" intact and operated after flight regenerated faster than control ones and showed long-lasting micro-"g" effect. The most recent experiment flew aboard the Bion-11 biosatellite. This test was performed for study on microgravity effect on neural retina regeneration after optic nerve lesioning in the newt. Obtained results confirmed our previous information about intensification of regenerative processes in detached neural retina in urodela exposed to simulated weightlessness (Grigoryan et al., 1998). In particular, we found the increase and activation of cell populations participating in neural retina restoration and maintenance of retinal structure. Our findings suggest that promoting effect of microgravity upon regeneration could be influenced by several factors, largely influenced by a response of the whole organism to changed gravity vector. We hypothesized the synthesis of the specific range of stress proteins induced by micro-"g" and their regulative role in cell proliferation. Such a hypothesis for the existence of "altered gravity stress proteins" is discussed. PMID- 12528723 TI - Re: K. Reddy et al.'s case report on a rare case of plasma cell vulvitis (Zoon's vulvitis). PMID- 12528724 TI - The impatient patient--reexamining difficult patient-provider relationships. PMID- 12528725 TI - Upholding standards of care for difficult patients. AB - Adverse encounters with patients can frustrate any physician. The dilemma arises, however, when physicians allow these frustrations to negatively affect their practice of medicine. Treating all patients respectfully should be the standard, but maintaining this standard is difficult for some physicians. PMID- 12528726 TI - Difficult relationships--interactions between family members and staff in long term care. AB - Staff of long-term care facilities and family members have a common responsibility to ensure the best course of treatment and everyday care for residents who often cannot speak for themselves. Understanding the difference between instrumental and preservative care, and who the proper agent is to provide care in each category will not only improve staff/family interactions, but residential care in general. The Resident Enrichment and Activity Program improves the family/staff relationship obliquely by involving family in social activities; the Family Involvement in Care program, and the Patterns in Caregiving program directly target the relationship and involve the facility's administration to effect policy change. PMID- 12528727 TI - Ultrastructural changes in osteocytes in microgravity conditions. AB - We examined the histology and morphometry of biosamples (biopsies) of the iliac crest of monkeys, flown 14 days aboard the "Bion-11", using electron microscopy. We found, that some young osteocytes take part in the activation of collagen protein biosynthesis in the adaptive remodeling process of the bone tissue to microgravity conditions. Osteocyte lacunae filled with collagen fibrils; this correlates with fibrotic osteoblast reorganization in such zones. The osteolytic activity in mature osteocytes is intensified. As a result of osteocyte destruction, the quantity of empty osteocytic lacunae in the bone tissue increases. PMID- 12528728 TI - The challenges of cross-cultural healthcare--diversity, ethics, and the medical encounter. AB - Difficulties in the provider-patient relationship arise from many sources, and pose various challenges to the integrity of the medical encounter. When these issues are especially sensitive or important to the patient's health and well being, a complete breakdown in the therapeutic relationship may result. The goal of the emerging field of cross-cultural healthcare is to improve providers' ability to understand, communicate with, and care for patients from diverse backgrounds. We should weave the concepts of cross-cultural care into the ethics of caring if we truly hope to have a positive impact on the health status of diverse patient populations. PMID- 12528729 TI - Guidelines for providing ethical care in difficult provider-patient relationships. PMID- 12528730 TI - Studies on clonogenic hemopoietic cells of vertebrate in space: problems and perspectives. AB - Hemopoietic tissues were studied in vertebrates launched aboard the Soviet (Russian) biosatellites ("Cosmos-1129, 1514, 1667, 1887 and 2044"; "Bion-10 and 11") between 1980 and 1996. In the bone marrow of rats exposed to spaceflight conditions, a statistically significant decrease in cell number was revealed in the progenitor cell compartment accounting for the compensatory response of granulocyte-macrophage (CFU-gm) and erythrocyte lineages (BFU-e and CFU-e) and in the compartment of multipotent hemopoietic stem cells (CFU-s), which is responsible for the permanent renewal of hemopoietic tissue. The number of stromal fibroblastic progenitors (CFC-f) in the bone marrow of these rats was also reduced. Apparently, changes in the hemopoietic stroma damage the hemopoietic microenvironment and, hence, may be responsible for changes observed in the hemopoietic tissue proper. Attempts were made to develop methods for analyzing morphologically indiscernible clonogenic hemopoietic cells of newts, and studies on the effects of spaceflight factors on these cells were performed. The results showed that the numbers of clonogenic cells in newts of the flight group newts were significantly lower than in control newts. The data obtained are used as the basis for formulating the problems to be studied, drawing up a program for further research on the effects of spaceflight factors on stem and other clonogenic hemopoietic cells, and developing new experimental models for analyzing stem cells, the state of the hemopoietic stroma, etc. PMID- 12528731 TI - The extent of written trial information: preferences among potential and actual trial subjects. AB - AIM: To investigate the preferred extent of written information in clinical trials among potential and actual trial participants. MATERIALS AND METHODS: Questionnaire survey among citizens of Copenhagen County (PUB, N=508), patients attending an out-patient clinic (OPC, N=200), and finally among participants in two clinical trials (ROC, N=32; MRCRUC, N=47--see Abbreviations). Questions concerned attitudes to and preferences towards a relatively short ("short form") and a more detailed information form ("long form") about a hypothetical, but realistic trial. RESULTS: Approximately 1/8 of the respondents in PUB were satisfied with the "short form", whereas this was the case for approximately 1/6 of outpatients and 1/5 of actual trial participants. Regarding the "long form" approximately three quarters of respondents in all groups were satisfied. Outpatients as a whole were satisfied to a larger extent that respondents from the PUB trial concerning the "short form" (p=0.04). The "long form" was preferred by a little less than 4/5 of respondents in all groups. CONCLUSION: Written information to trial subjects should be detailed, as a majority of both potential and actual research participants prefers this, given the choice between two information forms of different extent on the same trial. PMID- 12528732 TI - An ethical problem for discussion (transplantation research): a response to Stone and Fulbrook. PMID- 12528734 TI - Body shop: tighter regulations are needed if the tissue bank industry is to retain its credibility. PMID- 12528733 TI - Moral truthfulness in genetic counseling. PMID- 12528735 TI - Churches say no to gene patenting. PMID- 12528736 TI - Prenatal genetic testing in Japan. AB - OBJECTIVE: To examine the use of prenatal diagnosis and maternal serum marker tests (MSMTs) from January 1 to December 31, 1997, in Japan. METHODS: Questionnaires were sent to medical facilities, which represent about 80% of the facilities providing prenatal diagnosis. RESULTS: Prenatal diagnosis was performed in 5,748 women, of whom 5,538 (5,516 amniocenteses and 22 chorionic villus sampling; 96.3%) were screened for chromosomal abnormalities. 89.1% (221/248) of those diagnosed to have fetal abnormalities terminated the pregnancy. Written informed consent signed by both the obstetrician and the pregnant woman undergoing prenatal diagnosis was the policy in 76.5% facilities. A total of 14,682 MSMTs were performed: 8,919 were in pregnant women under 34 years of age and 5,763 were in women aged 35 years or older. Follow-up data were obtained in 10,209; 6,918 women under 34 years were tested; of those 448 had a positive result; 349 underwent amniocentesis and trisomy 21 was diagnosed in 9. 3,291 women aged 35 years or older were tested; of those 634 had a positive result; 607 underwent amniocentesis and trisomy 21 was found in 16 cases. Written informed consent before MSMTs was obtained in 34.5% of the facilities. CONCLUSION: The data showed that only 4% of pregnant women above age 35 years underwent amniocentesis in Japan, and a small proportion of pregnant women were being offered MSMTs. Prenatal testing should be carried out only on a voluntary basis. For this purpose, women must be adequately informed. PMID- 12528737 TI - Supportive housing for homeless people with severe mental illness. AB - Research suggests that as many as 110,000 single adults with severe mental illness (SMI) are homeless on any given day in the United States. The combination of mental illness and homelessness make this population especially hard to reach through either housing or mental health programs alone. Supportive housing programs, which provide independent housing along with health and social services, hold great promise for this population, but are costly to launch and maintain. This Issue Brief highlights a landmark study that examines the extent to which supportive housing costs are offset by reductions in the use of public services for health, corrections, and shelter. PMID- 12528738 TI - Papers in the programme "Assistance and people with HIV-AIDS". IV: ethical and moral aspects. PMID- 12528739 TI - Health care and society. A conflicting reality: the limits of technology. PMID- 12528740 TI - The risk of injury to children in compact pickup trucks. AB - Nearly one million compact pickup trucks were sold last year in the US. Manufacturers now produce extended-cab models of pickups such as the Ford Ranger, Chevrolet S-10, Dodge Dakota, and Toyota Tacoma that can accommodate at least two rear-seated passengers, making them attractive to families with children. However, the safety of these rear seats for children has not been determined. This Issue Brief summarizes a new study that examines and quantifies the risk of injury to children riding in compact pickup trucks. Based on these findings, parents should be advised against using these vehicles as family transportation. PMID- 12528741 TI - Health care and society. A conflicting reality: dehumanization in intramural and extramural health care, its underlying causes and expectations for the future. PMID- 12528742 TI - Health care and society. The illumination of this reality: light in inter religious dialogue. Judaism and health care. PMID- 12528743 TI - Health care and society. Action to be taken: the new frontiers. PMID- 12528744 TI - Tensions in antibiotic prescribing. AB - Since 1999, the Agency for Healthcare Research and Quality has funded seven centers across the country to provide practical guidance to physicians and other health care professionals about the drugs they prescribe. These Centers for Education and Research on Therapeutics (CERTs) develop, translate and disseminate objective information on drugs to improve practice. The University of Pennsylvania's CERTs focuses on developing evidence for optimal treatment strategies for infectious diseases, and promoting the judicious use of antibiotics to combat the problem of antibiotic resistance. This Issue Brief explores one of the fundamental challenges physicians face in optimizing antibiotic use: the potential conflict between what is best for an individual patient, and what is best for society as a whole. PMID- 12528745 TI - The human genome and the genetic supermarket. PMID- 12528746 TI - Can secular patients survive Catholic hospitals?: merger trends threaten abortion, contraception, living wills. PMID- 12528747 TI - Living with dementia: caregiver perspectives. AB - About four million Americans currently live with Alzheimer's disease (AD) or related forms of dementia. Because the disease process impairs language, insight, and judgment, family members become "caregivers." These caregivers, either in part or in full, often make decisions on patients' behalf. This Issue Brief summarizes a series of studies that describe how caregivers make decisions for AD patients, and caregiver perspectives on the quality of life for relatives with AD. PMID- 12528748 TI - Practicing medicine without a license: when church law shapes health care, patients suffer. PMID- 12528749 TI - The ex-abortionists: why they quit. PMID- 12528750 TI - Needlestick injuries to nurses, in context. AB - Injuries with used needles and other "sharps" put health care workers at risk for serious bloodborne infections, such as HIV and hepatitis B and C. To some extent, this risk can be lessened through safer techniques (such as not recapping needles) and safer devices (such as needleless and self-sheathing equipment). But these injuries occur within a context (often a hospital unit) with organizational features that may themselves contribute to an increased or decreased risk. This Issue Brief summarizes a series of studies that investigate whether workplace aspects of the hospital (such as staffing levels, and organizational structure and climate) affect the risk of needlestick injuries to nurses. PMID- 12528751 TI - The illusion of animal rights. PMID- 12528752 TI - The zygote and personhood. PMID- 12528753 TI - Under Oregon's iron shroud: real people, real deaths. PMID- 12528754 TI - Firearm injury in America. AB - In 2000, nearly 29,000 people in the U.S. died from firearm injury.The vast majority of these people died from suicide (58%) or homicide (38%). And for every person who died, at least two others were shot and survived, often with permanent disability. The Firearm Injury Center at Penn (FICAP), founded in 1997, is a unique collaboration among health professionals, researchers and communities to address the magnitude and impact of firearm injury and violence. In this Issue Brief, FICAP presents an overview of firearm violence, and discusses public health approaches to reducing the toll of violent injury. PMID- 12528755 TI - Fitness, fatness and activity as predictors of bone mineral density in older persons. AB - OBJECTIVES: To determine relationships of bone mineral density (BMD) with fitness, physical activity, and body composition and fat distribution. DESIGN: Cross-sectional. SETTING: General Clinical Research Center, Johns Hopkins Bayview Medical Center, Baltimore, Maryland. SUBJECTS: Men (n = 38) and women (n = 46), aged 55-75 years with high normal blood pressure or mild hypertension but otherwise healthy. METHODS: Aerobic fitness (oxygen uptake) on a treadmill, muscle strength by one-repetition maximum, activity by questionnaire, abdominal obesity by magnetic resonance imaging; anthropometrics, and body composition by dual energy X-ray absorptiometry (DXA) which measured total fat and lean mass, and BMD for the total skeleton, lumbar spine (L1-L4) and total hip. RESULTS: Aerobic fitness did not correlate with BMD. Using multivariate analysis to ascertain independent contributions to the variance in BMD, in women, with adjustment for hormone replacement therapy (HRT), total skeleton BMD was independently related to muscle strength and abdominal total fat; total hip BMD to body weight; lumbar spine BMD to abdominal total fat. HRT also influenced BMD in the lumbar spine. In men, lumbar spine BMD was independently related to abdominal total fat physical activity and total hip BMD related to lower body strength. P < 0.05 for all of these correlations. CONCLUSIONS: Abdominal obesity and muscle strength emerge as predominant correlates of BMD in older persons with stronger relationships seen in women. Body weight and HRT also explained portions of the variance in BMD in women. Whether abdominal obesity is simply a marker for general obesity or has independent protective effects on bone is yet to be determined. PMID- 12528756 TI - Search for occult secondary osteoporosis: impact of identified possible risk factors on bone mineral density. AB - OBJECTIVES: To determine whether the use of more elaborate diagnostic tests can identify possible risk factors for secondary osteoporosis and to evaluate the impact of these possible risk factors on the severity of bone disease in the study population. DESIGN: Cross-sectional study. SETTING AND PARTICIPANTS: We have investigated 377 subjects (285 females, 92 males) with osteoporosis (T-score less than -2.5 in dual energy X-ray absorption) or nontraumatic lumbar vertebral fractures; these patients were referred to our hospital, a secondary care centre, for evaluation and treatment of osteoporosis. RESULTS: Osteoporosis without attributable risk factor was diagnosed in 106 women (37%) and 30 men (33%). In 241 patients (179 women, 62 men) one or more possible risk factors for osteoporosis (in this paper also called subclinical disease) were revealed. The most common were lactose malabsorption, disturbed exocrine pancreatic function and renal tubular disturbances, including renal hypercalciuria, incomplete renal tubular acidosis and mild phosphate diabetes. The number of possible risk factors in the individual patient was significantly related to the severity of osteoporosis as assessed by Z-scores (Spearman correlation r = -0.43, P < 0.001, n = 172 for females; r = -0.28, P < 0.05, n = 65 for males). CONCLUSIONS: All the identified subclinical diseases would have remained undetected if the currently accepted guidelines for the investigation of patients with osteoporosis were applied. The statistically significant correlation between the number of identified possible risk factors and the severity of bone disease in the individual patient strongly suggests the pathogenetic significance of the identified subclinical diseases. It is yet to be shown, whether specific treatment of these subclinical diseases yields additional improvement of bone mass as compared with standard treatment of osteoporosis. PMID- 12528757 TI - Impact of smokeless tobacco use on smoking in northern Sweden. AB - BACKGROUND AND OBJECTIVES: For many years Swedish men have had the world's lowest rates of smoking and smoking-related mortality. Despite these facts, a thorough analysis of tobacco use patterns in Sweden has not been performed. The purpose of this study was to examine the prevalence and interaction of cigarette smoking and use of Swedish moist snuff (snus) in the population of northern Sweden. DESIGN: The study cohort of 2998 men and 3092 women aged 25-64 was derived from the northern Sweden MONICA study, consisting of population-based surveys in 1986, 1990, 1994 and 1999. Detailed information on tobacco use was used to develop prevalence data, and the prevalence ratio was used to compare rates amongst various subgroups. RESULTS: Amongst men ever-tobacco use was stable in all survey years at about 65%, but the prevalence of smoking declined from 23% in 1986 to 14% in 1999, whilst snus use increased from 22% to 30%. In women the prevalence of smoking was more stable in the first three surveys (approximately 27%) but was 22% in 1999, when snus use was 6%. In all years men showed higher prevalence of ex-smoking than women. A dominant factor was a history of snus (PR = 6.18, CI = 4.96-7.70), which was more prevalent at younger ages. CONCLUSIONS: The recent transition from smoking to snus use amongst men, and incipiently amongst women, in northern Sweden is remarkable and relevant to the global discussion on strategies to reduce smoking. PMID- 12528758 TI - The effect of streptokinase neutralizing antibodies on fibrinolytic activity and reperfusion following streptokinase treatment in acute myocardial infarction. AB - OBJECTIVES: To evaluate tissue plasminogen activator (tPA) activity as a measure of fibrinolytic response to treatment with streptokinase (SK) and to relate this to the effect of pretreatment SK antibodies and to successful reperfusion assessed by continuous computerized vectorcardiography (VCG). SETTING: Umea University Hospital. SUBJECTS: A total of 104 patients with acute myocardial infarction (AMI) treated with SK and no history of previous SK treatment were studied. The tPA activity was measured 4 h after the start of treatment. The effect of pre-existing neutralizing antibodies to SK was analysed with a functional assay in pretreatment samples. Reperfusion was evaluated with VCG. MAIN OUTCOME MEASURES: Successful reperfusion. RESULTS: Fifty-five patients (53%) were classified as successfully reperfused. The risk for failed reperfusion was calculated in logistic regression models. In a univariate model, a borderline significant increase in the risk of failed reperfusion was observed in intermediate levels of SK neutralizing antibodies, but not in the highest levels. In a multivariate model, only high tPA activity, >25 U mL(-1), at 4 h (OR 0.17: 95% CI: 0.06-0.51) was associated with a higher rate of reperfusion whilst longer time to treatment (OR 1.17; 95% CI: 1.02-1.35) was associated with a higher risk of failed reperfusion. There was no significant correlation between neutralizing antibodies to SK and tPA activity at 4 h. CONCLUSION: The SK treatment of AMI induced high levels of tPA activity which were associated with successful reperfusion. The effect of pre-existing SK antibodies had no significant influence on reperfusion and were not correlated to the fibrinolytic activity obtained. PMID- 12528759 TI - Telemedicine used for remote prehospital diagnosing in patients suspected of acute myocardial infarction. AB - INTRODUCTION: In patients with acute myocardial infarction (AMI), considerable time elapses from symptom onset until initiation of thrombolytic therapy or primary percutaneous coronary intervention. Prehospital diagnosing can reduce time delays, and remote diagnosing using telemedicine may be appropriate in areas and countries where ambulances are not staffed with physicians. OBJECTIVES: To evaluate whether it was technically feasible for physicians at a remote university hospital to diagnose ST-segment-elevation-AMI (AMI(STelev)) in patients suspected of AMI, who were transported by ambulances to a local hospital. To determine associated prehospital delays and in-hospital treatment delays. METHODS: Patients carried in telemetry equipped ambulances had 12-lead electrocardiograms (ECGs) acquired as soon as possible. En route to the local hospital the ECGs were transmitted to a remote university hospital, by use of the GSM-system. The physician on call at the university hospital interviewed the patients, who were provided with cellular phone headsets, and alerted the local hospital if signs of AMI(STelev), bundle-branch-block-AMI or malignant arrhythmia were detected. Patients transported by traditional ambulances were included in a prospective control group. RESULTS: In 214 (86%) of 250 patients prehospital diagnosing was successful. Geographically related transmission problems were the primary reason for failure. Ninety-eight per cent of transmitted electrocardiograms and obtained history takings were technically acceptable for diagnostic purposes. Door-to-needle times were shorter amongst patients with AMI(STelev) who were subjected to prehospital diagnosing (n = 13) as compared with patients transported by traditional ambulances (n = 14) (38 vs. 81 min) (P = 0.004). CONCLUSIONS: It was technically feasible to use telemedicine for remote prehospital diagnosing of patients suspected of AMI. Patients subjected to prehospital diagnosing had shorter door-to-needle times compared with a prospective control group. PMID- 12528760 TI - Enterovirus, mycoplasma and other infections as predictors for myocardial infarction. AB - OBJECTIVES: To study antibodies against five infectious agents for their prediction of major coronary events in men with and without evidence of coronary heart disease at baseline. DESIGN: A case-control study nested within a prospective population study. SUBJECTS: The study cases included 441 men 45-64 years old with nonfatal myocardial infarction or coronary death within a mean follow-up time of 10 years. A total of 165 men had already signs of heart disease at baseline, whilst 276 were apparently healthy at the beginning of the study. Two controls for each case were matched for age, heart disease status and place of residence. Antibodies against enterovirus, Mycoplasma pneumoniae, Chlamydia pneumoniae, cytomegalovirus and adenovirus were determined. RESULTS: Men without reported baseline heart disease, but not those with heart disease, showing the highest quartile of antibodies to enterovirus and mycoplasma or increased levels of immune complex-bound antibodies to chlamydia had a significantly higher risk of coronary events than men with lower level of antibodies. The increased risk demonstrated in men with high levels of antibodies to enterovirus and mycoplasma remained significant after adjustment for other antibodies, acute-phase reactant and conventional risk factors. Serological evidence of infection by multiple agents was also significantly associated with coronary events. CONCLUSIONS: Serological evidence for several infectious agents is associated with the risk of coronary heart disease, but only in men without baseline history of heart disease. PMID- 12528761 TI - Relationships between left ventricular mass and the renin-angiotensin system, catecholamines, insulin and leptin. AB - OBJECTIVES: Several neurohormonal systems have been suggested to stimulate myocardial cell growth, and thus take part in the development of left ventricular (LV) hypertrophy. We studied associations between LV mass and markers of the renin-angiotensin-aldosterone system (RAAS), the sympathetic nervous system, glucose homeostasis and leptin. DESIGN: A total of 115 hypertensive patients with LV hypertrophy and two age- and gender-matched control groups consisting of 38 hypertensive patients without LV hypertrophy and 38 normotensive subjects were included. We examined determinants of the RAAS, plasma levels and 24-h urinary excretions of noradrenaline and adrenaline, plasma proinsulin, insulin, glucose, leptin and LV mass by echocardiography. RESULTS: Plasma renin activity (PRA) and serum aldosterone were higher (both P < 0.001) in the LV hypertrophy group than in patients without LV hypertrophy and normotensive subjects (1.0 +/- 0.8, 0.2 +/ 0.2 and 0.2 +/- 0.2 ng mL(-1) h(-1), and 327 +/- 126, 269 +/- 146 and 221 +/- 80 pmol L(-1), respectively). PRA and aldosterone both related (P < 0.001) to LV mass index (r = 0.44 and 0.27, respectively). Catecholamine levels and excretions were similar in all three groups and did not relate to LV mass index. Proinsulin, insulin and leptin levels were all elevated in the hypertensive patients (P < 0.01), but proinsulin, insulin, insulin sensitivity (by the homeostasis model assessment) and leptin did not relate to LV geometry, when indexed for body size. CONCLUSIONS: Plasma renin activity and serum aldosterone levels are elevated in hypertensive LV hypertrophy and relate to LV mass index. In addition to blood pressure, activation of the RAAS may be an important nonhaemodynamic mechanism in the control of LV hypertrophy. PMID- 12528762 TI - Circulating oxidized low-density lipoprotein (LDL) is associated with risk factors of the metabolic syndrome and LDL size in clinically healthy 58-year-old men (AIR study). AB - OBJECTIVES: Hypothetically the atherogenic effect of the metabolic syndrome may be mediated through the increased occurrence of small LDL-particles which are easily modified to atherogenic oxidized LDL (ox-LDL). The aim of this study was to test this concept by examining the association between circulating ox-LDL, LDL particle size, and the metabolic syndrome. DESIGN AND RESULTS: A population-based sample of clinically healthy 58-year-old men (n = 391) was recruited. Ox-LDL was measured by ELISA (specific monoclonal antibody, mAb-4E6) and LDL-particle size by gradient gel electrophoresis. The results showed that ox-LDL significantly correlated to factors constituting the metabolic syndrome; triglycerides (r = 0.43), plasma insulin (r = 0.20), body mass index (r = 0.20), waist-to-hip ratio (r = 0.21) and HDL (r = -0.24); (P < 0.001). Ox-LDL correlated also to LDL particle size (r = -0.42), Apo-B (r = 0.70), LDL (r = 0.65); (P < 0.001) and, furthermore, with Apo A-1 (r = -0.13) and heart rate (r = 0.13); (P < 0.01). CONCLUSION: The metabolic syndrome was accompanied by high plasma ox-LDL concentrations compared with those without the syndrome. Ox-LDL levels were associated with most of the risk factors constituting the metabolic syndrome and was, in addition related to small LDL-particle size. To our knowledge the present study is the first one to demonstrate that circulating ox-LDL levels are associated with small LDL-particle size in a population representative sample of clinically healthy middle-aged men. The high degree of intercorrelation amongst several factors makes it difficult to clarify the independent role of any specific factor. PMID- 12528763 TI - The effects of exercise training on walking function and perception of health status in elderly patients with peripheral arterial occlusive disease. AB - OBJECTIVE: To determine the effects of 12-week exercise programme on ambulatory function, free-living daily physical activity and health-related quality of life in disabled older patients with intermittent claudication. DESIGN: Prospective, randomized controlled trial. SETTING: University Medical Center and Veterans Affairs Medical Center, Taipei, Taiwan. SUBJECTS: Thirty-two of 64 patients with Fontaine stage II peripheral arterial occlusive disease (PAOD) were randomized to exercise training and 32 to usual care control. Five patients from the exercise group and six patients from the control group dropped out, leaving 27 and 26 patients, respectively, completing the study in each group. INTERVENTIONS: Twelve weeks of treadmill exercise training. MAIN OUTCOME MEASURES: Treadmill walking time to onset of claudication pain and to maximal claudication pain, 6-min walk distance, self-reported ambulatory ability and perceived health-related quality of life (QOL). RESULTS: Compliance of exercise programme was 83% of the possible sessions. Exercise training increased treadmill walking time to onset of claudication pain by 88% (P < 0.001), time to maximal pain by 70% (P < 0.001), and 6-min walk distance by 21% (P < 0.001). SUBJECTS: Perception of health related QOL improved from 12% to 178% in the exercise group. These improvements were significantly better than the changes in the control group (P < 0.05). CONCLUSIONS: Significant improvements in claudication following 12-week exercise training in elderly PAOD patients were observed. Increase in treadmill walking time to maximal claudication pain in these patients translated into the improvement of perceived physical health, which enabled the patients to become more functionally independent. PMID- 12528764 TI - Homocysteine, malondialdehyde and endothelial markers in dialysis patients during low-dose folinic acid therapy. AB - OBJECTIVES: Haemodialysis patients have elevated levels of the atherogenic amino acid homocysteine. We wanted to assess the effects of small doses of intravenous folinic acid (the active form of folic acid) on some biochemical risk factors of cardiovascular disease. DESIGN: Longitudinal and open intervention study. SETTING: Two dialysis units in the County of Rogaland. SUBJECTS: All patients on maintenance haemodialysis were invited, and 32 of 35 patients gave their informed consent. INTERVENTIONS: After each dialysis session, the patients were given 1.0 mg of folinic acid intravenously thrice a week for a period of 3 months. Prior to and during the study, all patients were on maintenance supplementation with small doses of vitamins B1, B2, B3, B5, B6 and B12. MAIN OUTCOME MEASURES: Changes in the levels of (i) plasma total homocysteine (p-tHcy) and folate, (ii) circulating endothelium related proteins--markers of endothelial activation and (iii) serum malondialdehyde (S-MDA)--a marker of oxidative stress and lipid peroxidation. RESULTS: The p-tHcy levels were reduced by 37% (P < 0.0001), whilst the serum and erythrocyte folate levels increased by 95 and 104%, respectively (P < 0.0001 for both). The circulating levels of endothelium related cellular adhesion molecules and haemostatic factors remained high and unchanged, except the thrombomodulin (TM) levels increased (P = 0.0004). The high levels of S-MDA were reduced by 26% (P = 0.003). CONCLUSIONS: Low doses of folinic acid given intravenously to dialysis patients reduced their levels of p-tHcy and S-MDA and thus improved their cardiovascular risk profile. The concurrent increment in TM levels was unexpected and of unknown clinical significance. PMID- 12528765 TI - Determination of cardiac involvement in sarcoidosis by magnetic resonance imaging and Doppler echocardiography. AB - OBJECTIVES: To elucidate whether cardiac magnetic resonance imaging (MRI) could be useful in disclosing structural changes in the myocardium in sarcoidosis patients and to relate echo-Doppler derived indices of left ventricular function to electrocardiogram (ECG) findings. DESIGN: The MRI was performed in 18 consecutive patients with sarcoidosis. Left ventricular ejection fraction (LVEF), i.e. systolic function, was estimated echocardiographically by Simpson's two dimensional method (n = 16). Diastolic function was estimated by age-corrected Doppler-derived indices: isovolumetric relaxation time (IVRT), deceleration time (DT) and early filling/atrial contraction ratio (E/A ratio). RESULTS: Eleven patients had conduction defects or dysrhythmias (ECG+) whilst seven patients had a normal ECG (ECG-). In two patients, high signalling, contrast-enhanced, isolated regions, suggestive of deposits, were seen in the left ventricular myocardium on MRI. Both these patients had abnormal ECGs and signs of systolic and/or diastolic dysfunction on echocardiography. LVEF was subnormal in seven of 10 of the ECG+ patients and in two of six of the ECG-. Signs of diastolic dysfunction were found in 59% and 56% of the measurements in the ECG+ and ECG- patients, respectively. CONCLUSION: We conclude (i) that myocardial deposits on MRI in sarcoidosis patients have a high specificity for cardiac involvement but a rather low sensitivity; (ii) that a substantial proportion of sarcoidosis patients with abnormal ECGs have echocardiographic signs of systolic and/or diastolic dysfunction. PMID- 12528766 TI - Significance of tyrosine kinases in cancer: overview. PMID- 12528767 TI - Regulation and targets of receptor tyrosine kinases. AB - Ligand-mediated activation of receptor tyrosine kinases (RTKs) results in autophosphorylation of both the receptor catalytic domain and noncatalytic regions of the cytoplasmic domain. Catalytic domain phosphorylation leads to activation and potentiation of receptor kinase activity. Noncatalytic domain phosphorylation creates docking sites for downstream cytoplasmic targets, which bind to specific receptor phosphotyrosine residues. Downstream signaling pathways are constructed in a modular fashion. In addition to SH2 and PTB (phosphotyrosine binding) domains, downstream signal proteins also contain domains that recognize other protein and phospholipid motifs. The arrangement and re-arrangement of various combinations of modular domains in different signaling proteins (combinatorial use) has allowed for the creation of complex signaling networks and pathways. In addition to performing catalytic functions, signaling proteins serve as scaffolds for the assembly of multiprotein signaling complexes, as adaptors, as transcription factors and as signal pathway regulators. Recent results show that the juxtamembrane region of Eph receptors is important in receptor autoregulation. Mutations in the juxtamembrane region of several RTKs have been shown to play a role in oncogenesis. It is likely that dysregulation of other modular components of signaling pathways also plays a role in oncogenic transformation. PMID- 12528768 TI - Tyrosine kinases as targets for cancer therapy. AB - Enhanced protein tyrosine kinase (PTK) activity correlates with the development of cancer and other proliferative diseases. The hypothesis that PTK inhibitors may be of value in the treatment of cancer led to the systematic synthesis of selective tyrosine phosphorylation inhibitors (tyrphostins) that show in vitro and in vivo anticancer activity. This review will provide an overview of research efforts in the development of tyrphostins such as AG 957, AG 1112, and AG 1318. Other tyrphostins discussed are AG 1478 and RG 13022, which are both epidermal growth factor receptor kinase inhibitors; AG 490, a Jak-2 kinase inhibitor; AG 1296, a PDGFR kinase inhibitor; and STI 571 (imatinib, Glivec/Gleevec; Novartis Pharma AG, Basel, Switzerland). STI 571 is now approved for the treatment of chronic myeloid leukemia and shows activity against gastrointestinal stromal tumors. The chemistry, kinetics, biological activity, and clinical potential of these compounds will be discussed. PMID- 12528769 TI - Imatinib: a selective tyrosine kinase inhibitor. AB - The understanding of the pathophysiology of a large number of cancer types provides a strategy to target cancer cells with minimal effect on normal cells. Protein phosphorylation and dephosphorylation play a pivotal role in intracellular signaling; to regulate signal transduction pathways, there are approximately 700 protein kinases and 100 protein phosphatases encoded within the human genome. In cancer, as well as in other proliferative diseases, unregulated cell proliferation, differentiation and survival frequently results from abnormal protein phosphorylation. Although it is often possible to identify a single kinase that plays a pivotal role in a given disease, the development of drugs based upon protein kinase inhibition has been hampered by unacceptable side effects resulting from a lack of target selectivity. With the growing understanding of the molecular biology of protein tyrosine kinases and the use of structural information, the design of potential drugs directed towards the bind adenosine triphosphate (ATP)-binding site of a single target has become possible. These advances have transferred emphasis away from the identification of potent kinase inhibitors and more towards issues of target selectivity, cellular efficacy, therapeutic effectiveness and tolerability. In this paper, the relationship between molecular biology and drug discovery methods, as utilized for the identification of anticancer drugs, will be illustrated. PMID- 12528770 TI - Pharmacology of imatinib (STI571). AB - Deregulation of protein kinase activity has been shown to play a central role in the pathogenesis of human cancer. The molecular pathogenesis of chronic myelogenous leukemia (CML) in particular, depends on formation of the bcr-abl oncogene, leading to constitutive expression of the tyrosine kinase fusion protein, Bcr-Abl. Based on these observations, imatinib was developed as a specific inhibitor for the Bcr-Abl protein tyrosine kinase. The expanding understanding of the basis of imatinib-mediated tyrosine kinase inhibition has revealed a spectrum of potential new antitumor applications beyond the powerful activity already reported in the treatment of CML. Imatinib has shown activity in vivo against PDGF-driven tumor models including glioblastoma, dermatofibrosarcoma protuberans and chronic myelomonocytic leukemia. Antiangiogenic effects have been demonstrated by inhibition of PDGF-, VEGF (vascular endothelial growth factor)- and bFGF- (basic fibroblast growth factor) induced angiogenesis in vivo, and by inhibition of angiogenesis and tumor growth in an experimental bone metastasis model. Imatinib has been shown to reduce interstitial fluid pressure in an experimental colonic carcinoma model by blocking PDGF-mediated effects on tumor associated blood vessels and stromal tissue. It is also a potent inhibitor of the Kit receptor tyrosine kinase, and has demonstrated activity clinically against the Kit-driven gastrointestinal stromal tumor (GIST) and experimentally in small cell lung cancer cell lines. The pharmacology of imatinib and its activity in various tumor models is discussed. PMID- 12528771 TI - Clinical presentation of gastrointestinal stromal tumors and treatment of operable disease. AB - Gastrointestinal stromal tumors (GISTs) are generally found in the stomach or small intestine and less commonly in the colon, rectum or an intra-abdominal site. The patients symptoms on presentation are most commonly gastrointestinal bleeding. Surgery remains the standard treatment for nonmetastatic GISTs, but rates of disease recurrence are significant--5% in primary disease and 90% in locally advanced disease. Five-year survival following surgical resection varies between 35% and 65% on the basis of several published studies. Clinical knowledge of the prognosis of patients with GISTs remains rather limited--small tumor size, low-grade mitotic index and stomach location are factors associated with a more favorable prognosis. PMID- 12528772 TI - Pathology and diagnostic criteria of gastrointestinal stromal tumors (GISTs): a review. AB - Gastrointestinal stromal tumor (GIST) is the designation for the specific c-kit expressing and Kit-signaling driven mesenchymal tumors, many of which have Kit activating mutations. The specific identification of GIST has become increasingly important because a Kit-selective tyrosine kinase inhibitor, imatinib (Glivec, formerly known as STI571, Novartis Pharma AG, Basel, Switzerland), has shown promise as an effective adjuvant therapy treatment. GISTs are the most common mesenchymal tumors of the gastrointestinal (GI) tract. We estimate the frequency of malignant GISTs as 20% to 30% of the frequency of all soft-tissue sarcomas, but small benign tumors, often found incidentally during unrelated surgery or autopsy, are probably much more common. Older adults are most at risk for GIST; very rarely, GIST occurs in children and young adults (sometimes connected with Carney's triad), or on a familial basis. GISTs have been documented in all parts of the GI tract. A great majority of them occur in the stomach (60% to 70%) and small intestine (25% to 35%), with rare occurrence in the colon and rectum (5%), esophagus (<2%) and appendix. Some GISTs are primary in the omentum, mesentery or retroperitoneum, and are unrelated to the tubular GI tract. GISTs can be histologically identified as highly cellular spindle cell or epithelioid mesenchymal tumors, and morphology is somewhat site-dependent. However, common to all these tumors is expression of Kit (CD117 antigen), which is a major diagnostic criterion. Few other Kit-positive mesenchymal tumors of the GI tract are likely to be confused with GISTs; exceptions are metastatic melanoma and related tumors and malignant vascular tumors. Additional diagnostic criteria include common positivity for CD34 (70%), variable expression of smooth muscle actins (20% to 30%) and S100 protein (10%) and almost uniform negativity for desmin (only 2% to 4% of GISTs are positive). Although the prediction of malignancy in this tumor group is notoriously difficult, tumors that have mitotic activity counts exceeding 5 per 50 high power fields (HPF) or those larger than 5 cm have a high frequency of intra-abdominal recurrence and liver metastasis. In contrast, tumors smaller than 2 cm and those with mitotic activity counts <5 per 50 HPF are likely to be benign. These diagnostic criteria leave an inevitable gray area in the separation of benign and malignant tumors. Kit-activating mutations can be detected in at least 60% to 70% of GIST cases. Most of the mutations, in-frame deletions of several codons, are located in the juxtamembrane domain (exon 11) of the gene. Less commonly, mutations have been detected in the extracellular domain (exon 9), and tyrosine kinase domains (exons 13 and 17). Functional analysis of the different c-kit mutations and their impact on the response to tyrosine kinase inhibitors are under intense investigation. PMID- 12528773 TI - Identification and treatment of chemoresistant inoperable or metastatic GIST: experience with the selective tyrosine kinase inhibitor imatinib mesylate (STI571). AB - Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal neoplasms of the gastrointestinal tract. The concept of GIST and the definition of GIST pathology have evolved greatly over the past 5 years. GIST has been shown to share immunohistochemical, ultrastructural and histogenic similarities with the interstitial cells of Cajal. Both GIST and the interstitial cells of Cajal express KIT, the receptor tyrosine kinase that is the protein product of the c kit proto-oncogene. KIT is universally phosphorylated in GISTs. Sequencing of c kit complementary DNA from human GIST cells has demonstrated a high frequency of mutations that lead to constitutive activation of the KIT tyrosine kinase in the absence of stimulation by its physiologic ligand (stem cell factor). This, in turn, causes uncontrolled stimulation of downstream signaling cascades with aberrant cellular proliferation and resistance to apoptosis. Historically, malignant GIST has been highly refractory to conventional cytotoxic therapy. Signal transduction inhibition as cancer therapy was first tested successfully with imatinib mesylate (formerly known as STI571), a selective small-molecule tyrosine kinase inhibitor, with the initial target being blockade of Bcr-Abl, the oncogene with tyrosine kinase activity responsible for the pathogenesis of chronic myelogenous leukemia (CML). Imatinib was subsequently shown to block activity of the KIT tyrosine kinase as well, and in laboratory studies this led to apoptotic death of GIST cells. The first GIST patient to receive imatinib exhibited dramatic benefit despite far-advanced metastatic disease that was previously refractory to all chemotherapy. Subsequently, multicenter clinical trials have been performed to assess the safety, efficacy and biologic activity of imatinib in patients with advanced GIST. The results from these studies have established imatinib as an effective new therapeutic alternative for the majority of patients with advanced GIST, a solid tumor for which no prior chemotherapy has ever shown antitumor efficacy. This work provides proof of concept to the hypothesis that selective inhibition of aberrant signal transduction can provide important anticancer activity, if the proper signaling pathways are identified and blocked. PMID- 12528774 TI - Use of positron emission tomography in oncology and its potential role to assess response to imatinib mesylate therapy in gastrointestinal stromal tumors (GISTs). AB - The reliability of established anatomical imaging techniques, such as computed tomography (CT) and magnetic resonance imaging (MRI), is compromised in following response to certain types of treatment if metabolic improvement occurs before morphologic change is apparent. Thus, traditional imaging techniques cannot discriminate early tumor response because they are based on purely visual structural assessments. Recently, the use of positron emission tomography (PET), most commonly employing the radiotracer 18F-fluoro-2-deoxy-D-glucose (FDG), has been shown to improve the assessment of tumor behavior by highlighting early functional changes in tumor glucose metabolism that appear to correlate closely with metabolic tumor response to imatinib mesylate. Like CT and MRI, PET can identify an abnormal mass; its improvement over these techniques lies in its ability to differentiate active tumor from necrosing tissue, malignant from benign tissue, and recurrent tumor from scar tissue. Understanding and using this tool should improve our ability to accurately follow response in GIST patients treated with imatinib mesylate, and permit this new therapeutic approach to be used optimally with accurate follow-up assessments and informed therapeutic decision-making. PMID- 12528775 TI - Overview of issues related to imatinib therapy of advanced gastrointestinal stromal tumors: a discussion among the experts. AB - Information regarding the activity of imatinib in patients with metastatic gastrointestinal stromal tumors (GISTs) has accumulated rapidly. Nevertheless, several important issues about imatinib therapy as well as these tumors themselves remain to be answered. Importantly, the optimal dose and duration of imatinib therapy are unknown, with daily doses of 400 mg and 600 mg producing comparable response rates in a phase II study. Moreover, the role of surgery following maximal responses to imatinib and those of functional imaging and use of biopsies in monitoring treatment responses need to be investigated. Further understanding of the molecular and pathologic characteristics of GISTs that are responsive or resistant to imatinib is also needed. This paper summarizes a symposium that was held in Helsinki, Finland, in September 2001. PMID- 12528776 TI - Imatinib and chronic myeloid leukemia: validating the promise of molecularly targeted therapy. AB - The Bcr-Abl tyrosine kinase inhibitor imatinib (Glivec, formerly STI571, Novartis Pharma AG, Basel, Switzerland) produces complete hematologic and cytogenetic responses in a substantial percentage of chronic myeloid leukemia patients. Imatinib is effective in chronic phase, accelerated phase and blast crisis, with lower response rates in patients with more advanced disease. Although responses have been durable in chronic phase patients, relapses have been common in blast crisis. Relapse has been associated with reactivation of Bcr-Abl kinase activity. The clinical development of imatinib illustrates the effectiveness of targeting molecular pathogenetic events. Hopefully, this example can be extended to other malignancies. PMID- 12528777 TI - Imatinib: the first 3 years. AB - Imatinib (Glivec, formerly STI571, Novartis Pharma AG, Basel, Switzerland) potently inhibits several protein tyrosine kinases, including Bcr-Abl, Kit, and the platelet-derived growth factor receptor. Phase I and II studies demonstrated that orally administered imatinib is highly effective and well tolerated in all phases of chronic myeloid leukemia (CML) at doses ranging from 400 to 600 mg. Importantly, preliminary evidence suggests that patients with advanced CML achieving hematologic or major cytogenetic responses to imatinib may have longer survival than those without such responses, whereas chronic phase patients who respond to treatment may have longer times to disease progression. Ongoing and planned studies are focused on optimizing CML treatment with imatinib, evaluating imatinib-based combination therapy, defining additional therapeutic targets and exploring the use of imatinib in children. In particular, results from several combination phase I studies are expected shortly, including an evaluation of combination imatinib-interferon-alpha therapy and imatinib-cytarabine in chronic phase CML, and a phase I study of single-agent imatinib in children with Philadelphia chromosome-positive leukemia is ongoing. A large phase III trial comparing imatinib with standard inferferon alfa plus cytarabine in first-line CML treatment is also ongoing. PMID- 12528778 TI - Update of phase I study of imatinib (STI571) in advanced soft tissue sarcomas and gastrointestinal stromal tumors: a report of the EORTC Soft Tissue and Bone Sarcoma Group. AB - In a phase I study conducted by the EORTC Soft Tissue and Bone Sarcoma Group, 40 patients with advanced soft tissue sarcomas, most of whom had gastrointestinal stromal tumors (GISTs), received imatinib at doses of 400 mg q.d., 300 mg b.i.d., 400 mg b.i.d., or 500 mg b.i.d. Dose-limiting toxicities, including severe nausea, vomiting, edema and rash, were seen at the highest dose level; the maximum tolerated dose was therefore 400 mg b.i.d. Imatinib was active in the group of 35 patients with GISTs, producing partial responses in 19 (54%) patients and stable disease in 13 patients (37%). Responding patients have now been followed for a minimum of 10 months. The most common side effects seen in patients continuing on therapy have been periorbital edema (40%), peripheral edema (37.5%), fatigue (30%), skin rash (30%) and nausea/vomiting (25%). Severe late myelosuppression has also been seen occasionally. Eighteen (51%) GIST patients continue to have partial responses and 11 (31%) continue with stable disease. Thus, 82% of patients with GISTs are still obtaining clinically important benefits with continued imatinib therapy. Some patients showed accelerated progressive disease shortly after starting imatinib. On the other hand, following drug withdrawal, 2 patients had reductions in tumor burden and remain alive without drug therapy. In summary, imatinib is generally well tolerated and has significant activity during long-term treatment of patients with advanced GISTs. PMID- 12528779 TI - Epidemiology and prevention of childhood injuries. AB - Traumatic injury is the leading cause of death in children after infancy. The leading causes of childhood injury deaths are motor vehicle crashes, submersion injury, homicide, suicide, and fires. Injuries are not random events. Factors associated with injuries allow identification of high-risk populations and targeted interventions. Injury research includes development of conceptual models to include preinjury, event, and postevent features that can be modified to prevent or limit injuries. Successful prevention strategies often include multifaceted approaches such as education, incentives for safe human behavior, legislation/enforcement, and environmental changes. Preventive programs must weigh both societal and economic values and costs. Careful evaluation for effectiveness of injury prevention programs to decrease or limit injury continues to be a challenge. Generally, passive measure such as improved engineering are more effective than measures that require modification of human behaviors. Childhood injury prevention programs have reduced deaths from some causes such as motor vehicle crashes, but deaths from gun-related homicide and suicide remain high. Critical care providers can actively engage in both prevention efforts and improved acute care of the severely injured child. PMID- 12528780 TI - Critical care management of head trauma in children. AB - Trauma is the leading cause of both morbidity and mortality in the pediatric population, and traumatic injury causes > 50% of all childhood deaths. Significant mortality rates have been reported for children with traumatic brain injury. Although children have better survival rates as compared with adults with traumatic brain injury, the long-term sequelae and consequences are often more devastating in children due to their age and developmental potential. The costs involved in the care of a child with severe traumatic brain injury, extended over that child's lifetime, are significant. It is unfortunate that despite preventive measures, traumatic brain injury remains the major morbidity and mortality factor for children. PMID- 12528781 TI - Submersion and asphyxial injury. AB - Drowning and other asphyxial injuries are important causes of childhood morbidity and mortality. In this review, the epidemiology, pathophysiology, and treatments applied to near-drowning victims are discussed, with an emphasis on the difficulties encountered attempting to predict outcome using current methods. PMID- 12528782 TI - Pediatric thoracic trauma. AB - Although thoracic injuries occur less frequently in children than adults, they remain a source of substantial morbidity and mortality. Disparate problems such as rib fractures, lung injury, hemothorax, pneumothorax, mediastinal injuries, and others may present in isolation or in combination with one another. Knowledge of the manner in which pediatric anatomy, physiology, and injury patterns change with age may expedite the evaluation of the pediatric chest after trauma. Differences in pulmonary functional residual capacity, blood volume, chest wall and spinal soft-tissue mobility, and cardiac function may translate into problems or benefits of important consequence. For example, although more predisposed to hypoxemia, young children may remain well compensated hemodynamically, despite significant blood loss. Rare injuries in children, such as cardiac and great vessel trauma, may remain undiagnosed precisely because of their scarcity and protean symptoms. PMID- 12528783 TI - Abdominal and pelvic trauma in children. AB - Trauma is the leading cause of death and disability in children. More than 90% of pediatric trauma admissions are the result of a blunt mechanism. Although injury to the abdomen and pelvis account for only 10% of injuries sustained by victims of pediatric trauma, they can be potentially life threatening. Optimal evaluation of the injured child may require the use of multiple diagnostic modalities. The spleen is the most frequently injured intra-abdominal organ, followed by the liver, intestine, and pancreas. Fortunately, the majority of injuries to the spleen and liver can be treated nonoperatively. Conversely, injuries involving the intestine and pancreas often require operative intervention. PMID- 12528784 TI - Electrical injuries. AB - Electrical injury is a relatively infrequent but potentially devastating form of multisystem injury with high morbidity and mortality. Most electrical injuries in adults occur in the work-place, whereas children are exposed primarily at home. In nature, electrical injury occurs due to lightning, which also carries the highest mortality. The severity of the injury depends on the intensity of the electrical current (determined by the voltage of the source and the resistance of the victim), the pathway it follows through the victim's body, and the duration of the contact with the source of the current. Immediate death may occur either from current-induced ventricular fibrillation or asystole or from respiratory arrest secondary to paralysis of the central respiratory control system or due to paralysis of the respiratory muscles. Presence of severe burns (common in high voltage electrical injury), myocardial necrosis, the level of central nervous system injury, and the secondary multiple system organ failure determine the subsequent morbidity and long-term prognosis. There is no specific therapy for electrical injury, and the management is symptomatic. Although advances in the intensive care unit, and especially in burn care, have improved the outcome, prevention remains the best way to minimize the prevalence and severity of electrical injury. PMID- 12528785 TI - Pediatric orthopedic trauma: principles in management. AB - Optimal management of pediatric orthopedic trauma requires understanding the unique characteristics of the immature musculoskeletal system. The principles of pediatric orthopedic trauma management include anticipating future growth and bone remodeling potential, minimizing physeal injury, and aggressive treatment of open fractures and compartment syndromes. This article discusses the fundamentals of the pediatric musculoskeletal system and pediatric orthopedic trauma management in general and as they apply to specific injuries. PMID- 12528786 TI - Digital replantation in children. AB - Digital amputations in children can frequently be reconstructed using microvascular replantation techniques. An adequate preoperative assessment of the patient and amputated part is important to define associated trauma and to determine the candidacy for replantation. Preoperative care of the injured extremity and patient, as well as optimal perioperative medical and surgical management, increases the success of these microsurgical procedures. PMID- 12528787 TI - Pediatric trauma systems. AB - The origins of trauma systems in the United States date to the 1960s when physicians returning from wars abroad realized that lessons learned from managing military casualties could be applied to civilian traumatic injury. Over the next several decades, trauma centers and then trauma systems began to be developed in an attempt to improve prehospital and acute care for these patients. Although studies of trauma system effectiveness are fraught with methodologic difficulties, several types of studies (panel reviews of preventable deaths, registry studies, and population-based studies), suggest that there may be improvements in mortality when trauma systems are established. Further study needs to focus on other outcomes than mortality, such as return to function after rehabilitation. Pediatric trauma systems have by necessity developed within the "adult" systems in place. The history of pediatric system development and studies assessing outcomes are also discussed. Continued system development, assessment, and educational efforts about how childhood injuries are different are essential to combat this leading killer of children. PMID- 12528788 TI - Triage scoring systems, severity of illness measures, and mortality prediction models in pediatric trauma. AB - Trauma triage scores, severity of illness measures, and mortality prediction models quantitate severity of injury and stratify patients according to a specified outcome. Triage scoring systems are typically used to assist prehospital personnel determine which patients require trauma center care, but they are not recommended as the sole determinant of triage. Severity of illness measures and mortality prediction models are used in clinical and health services research for risk-adjusted outcomes analyses and institutional benchmarking. As clinicians and researchers, it is imperative that we be knowledgeable of the methodologies and applications of these scoring and risk prediction systems to ensure their quality and appropriate utilization. PMID- 12528789 TI - Multiple trauma in children: critical care overview. AB - Multiple trauma is more than the sum of the injuries. Management not only of the physiologic injury but also of the pathophysiologic responses, along with integration of the child's emotional and developmental needs and the child's family, forms the basis of trauma care. Multiple trauma in children also elicits profound psychological responses from the healthcare providers involved with these children. This overview will address the pathophysiology of multiple trauma in children and the general principles of trauma management by an integrated trauma team. Trauma is a systemic disease. Multiple trauma stimulates the release of multiple inflammatory mediators. A lethal triad of hypothermia, acidosis, and coagulopathy is the direct result of trauma and secondary injury from the systemic response to trauma. Controlling and responding to the secondary pathophysiologic sequelae of trauma is the cornerstone of trauma management in the multiply injured, critically ill child. Damage control surgery is a new, rational approach to the child with multiple trauma. The selection of children for damage control surgery depends on the severity of injury. Major abdominal vascular injuries and multiple visceral injuries are best considered for this approach. The effective management of childhood multiple trauma requires a combined team approach, consideration of the child and family, an organized trauma system, and an effective quality assurance and improvement mechanism. PMID- 12528790 TI - Pediatric trauma: postinjury care in the pediatric intensive care unit. AB - Traumatic injuries occur in > 20 million children each year and are the leading source of death in children over the age of 1 yr. Mechanisms of injury and subsequent therapies for critically injured children are diverse. This review will focus on resources and management strategies for caring for the severely injured child in the pediatric intensive care unit. PMID- 12528791 TI - Spinal cord injury. AB - Pediatric spinal cord injury is a relatively uncommon problem, responsible for approximately 5% of all spinal cord injuries. Anatomic and behavioral differences between adults and children lead to variation in injury type and severity. Young children are more prone to high cervical injuries, with nearly 80% of injuries in children < 2 yrs old affecting this area. As the child approaches 8-10 yrs of age, the spinal anatomy and therefore injury pattern more closely approximates adult injuries. Although the prevalence of spine injuries is lower in children, clearing the spine becomes more complex due to radiographic differences and the inability to "clinically" clear the cervical spine in young children. In this article, the types of injuries seen in children are discussed, with an emphasis on acute management and clearance of the cervical spine. Treatment options and long-term issues are also discussed. PMID- 12528792 TI - Burns. AB - During the past 20 yrs, as burn care has evolved as a specialty of surgery, survival and outcome quality have soared. Public expectations for survival and long-term outcomes are at previously unprecedented levels. These changes are the result of a number of advances in aspects of burn care that have occurred in parallel and have fostered increasing regionalization of this resource-intensive activity into fewer specialized centers. These are complex hospitalizations and can be divided into four phases: initial evaluation and resuscitation, initial wound excision and biological closure, definitive wound closure, and rehabilitation and reconstruction. PMID- 12528793 TI - Child abuse in the pediatric intensive care unit. AB - Critically ill and injured children due to abusive or inflicted injury represent a growing challenge for pediatric intensive care unit personnel in terms of the number of patients seen each year in the United States and the intellectual and emotional response required to deal with this tragic problem. We present a distillation of the current knowledge of childhood physical abuse with a focus on the child with inflicted injury who is admitted to the pediatric intensive care unit. In addition to a discussion of the epidemiology, clinical presentation, an approach to diagnosis, and treatment strategies, we also explore the legal issues that confront pediatric intensive care unit physicians in relation to determination of brain death, suitability of victims for organ donation, and the physician's role in the criminal investigation of child abuse and as a witness for court proceedings. PMID- 12528794 TI - Luminescence applications for chemotherapeutic drug development. AB - The application of in vitro and in vivo ATP bioluminescence systems as an integrated approach for preclinical research and development of new chemotherapeutic drugs is described. This approach includes both (a) the in vitro tumor response assay (TRA) system that utilizes new technologies for cell culture and ATP measurement of clinical specimens and (b) the use of human tumor cell lines transfected with Photinus pyralis luciferase (luc) gene for both in vitro and in vivo studies. Dried reagent microplates for TRA culture and counting procedures are described for a two-stage TRA method, which can be used to evaluate drug sensitivity and resistance of cells from clinical specimens after initial drug exposure in vitro. The use of dried reagent counting plates for screening and testing of agents against tumor cell lines is described, as well as an alternative method for screening and testing chemotherapeutic drugs in vitro with luc-transfected human tumor cell lines. The potential application of luc transfected reporter cell lines for in vivo studies of drug activity with photon imaging for analysis is discussed. PMID- 12528795 TI - The use of the MTT assay to study drug resistance in fresh tumour samples. AB - We have found the short-term MTT assay to be a simple, reproducible chemosensitivity technique, suitable for use throughout the time course of disease. We now have 12 years' experience of using this method in a variety of tumour types, both haematological and solid malignancies. Tumour cells are isolated from bone marrow, malignant effusions or solid biopsies and subjected to drug exposure for 48-96 h. Cell survival is measured by re-incubation in MTT for 4 h. We have found a significant correlation of in vitro results with in vivo outcome for acute myeloid leukaemia (AML) and for ovarian cancer (both p < 0.0001) with an assay sensitivity of 98% for AML and 81% for ovarian cancer. Furthermore, the 5-year survival of ovarian cancer patients treated with a drug found sensitive in vitro is significantly higher than that for patients treated with a drug found resistant in vitro (p = 0.033). We have correlated assay results with drug resistance markers. For example, expression of the newly described half transporter BCRP is related to daunorubicin resistance (p < 0.05). The MTT assay is also suitable for screening for modulation of drug resistance. We have found that the DNA polymerase inhibitor aphidicolin markedly increases in vitro sensitivity to the platinum drugs in ovarian cancer and cytosine arabinoside in AML in the majority of patients. The greatest effect was seen for patients deemed resistant in vitro to these agents. We have identified novel drug combinations which demonstrate significant synergism using this methodology and have also used it to study the emergence of drug resistance in cell line models with a view to its prevention. In conclusion, we have found the MTT assay to be a simple, repeatable, adaptable technique which produces accurate information to help the clinician select suitable treatment for individual cases. PMID- 12528796 TI - Chemosensitivity testing of human tumors using Si-sensor chips. AB - Chemotherapy is, on its own or in combination with other treatments, a very effective anticancer therapy. Introduced in the middle of the last century, chemotherapy today still faces the problem of determining which specific agent or agents are able to yield the desired clinical therapeutical effect for a particular tumor and patient. Numerous tests in vitro have been developed to detect chemosensitivity and chemoresistance and also for screening new drugs. Three groups of tests can be defined: 1, cell viability tests; 2, measurements of cell metabolism; and 3, clonogenic assays. Test time, tissue preparation, complexity of test performance, and correlation with the clinical progress of the disease are criteria used to judge how successful the tests are. The introduction of Si-sensor chips, which are able to detect metabolic changes in living cells, has opened up new possibilities in this field. Basically two sensor principles or types can be considered: (a) the light-addressed potentiometric sensor (LAPS) and (b) the multisensor array (MSA). Whereas LAPS measures one, MSA registers online many parameters (for instance, impedance, pH, O2, temperature). The aim of this chapter is to review this technology and to present recent applications using cells, tissue slices, and biopsies. PMID- 12528797 TI - Multiparametric sensor chips for chemosensitivity testing of sensitive and resistant tumor cells. AB - Many different assays have been developed for testing the chemosensitivity of tumor cells in vitro, usually based on a single biochemical or cellular parameter. A multiparametric test system has been developed that accommodates on a single chip numerous sensors for metabolic parameters, deltapH and deltapO2, as well as for morphological changes. The cells grow directly on the chips and can be continuously monitored online up to several days. The effects of various chemotherapeutic drugs on the metabolic profile of several tumor cell lines have been investigated. In colon carcinoma-derived LS174T cells, cytochalasin B markedly increased oxygen consumption while decreasing the rate of extracellular acidification. These effects, which reflect the biochemical action of cytochalasin B, were reversible on drug removal. In contrast, chloroacetaldehyde markedly reduced respiration, which recovered when the drug was removed. Primary breast cancer cells also responded to chloroacetaldehyde with a marked reduction in deltapO2, followed by a reduced rate of acidification. Comparing the metabolism of doxorubicin-sensitive and -resistant mouse sarcoma S180 cells, the rates of acidification and respiration were inhibited by doxorubicin only in the sensitive cells, whereas in the resistant cells oxygen consumption even increased. These examples demonstrate that this chip-based test system provides rapid and important information for assessing chemosensitivity of tumor cells. PMID- 12528798 TI - Development of a new in vitro chemosensitivity test using collagen gel droplet embedded culture and image analysis for clinical usefulness. AB - The results of chemotherapy are not fully satisfactory in many cases, particularly solid cancers. Therefore, it seems useful if the effective anticancer drugs can be selected for each patient using the screening methods. In such a background, we developed a new anticancer-drug sensitivity testing method that overcame several defects in the existing method, which is the collagen gel droplet embedded culture drug sensitivity test (CD-DST) and satisfies the following requirements: a high success rate, ability to assay biopsy specimens, and quantification of the anticancer effects without contamination with fibroblasts. Under the biological exposure condition, the sensitivities of the cancers were similar to their clinical response rate and there was a statistically significant correlation between clinically reported response rates and the response rates obtained by CD-DST (P < 0.01). Furthermore, the true positive rate was 79.8%, and the true negative rate was 88.8%. Sensitivity and specificity were 88.2% and 80.6%, respectively, resulting in an overall predictive accuracy of 84.1% (154/183). The CD-DST not only shows high predictive accuracy for humans, but because of the high correlation between the results of in vitro and nude mice assays, if it also is practical as a pre-clinical screening, it can easily provide these predictions. PMID- 12528799 TI - In vitro chemosensitivity testing of hematological cancer patients: detection of ornithine decarboxylase. AB - The development of reliable methods for the in vitro testing of sensitivity of cancer cells to various drugs has been a longstanding objective in cancer research and treatment Early attempts to develop individualized chemotherapy were based on clonogenic assays. These attempts failed because of low plating efficiencies. Nonclonogenic assays, such as the MMT test or ATP determinations, are based on metabolic activities and do not reflect the ability of cells to proliferate. To detect proliferation, we selected a universal marker--ornithine decarboxylase (ODC), which is expressed early in the cell cycle and has a short half-life. This marker was detected in hematological cancer cells by quantitative immunohistochemical analyses using an ODC antibody and a FITC-linked second antibody. Drug resistance was detected in five patients, who subsequently died. Lymphocytes from normal individuals were sensitive to all drugs tested, whereas 33 leukemia and lymphoma patients showed different sensitivities to certain drugs. The method also permitted testing of the effect of new drugs on the proliferation of lymphocytes from hematological cancer patients. This test is sensitive, and 100-1,000 cells are required per assay, which can be completed within 2 days. It is very likely that the assay could also be used to test solid tumor patients. PMID- 12528800 TI - The chemosensitivity profile of retinoblastoma. AB - Retinoblastoma is a rare malignant tumour of the developing retina with an incidence of 1 in 20,000 live births in all human races. Chemotherapy is used in retinoblastoma as adjuvant therapy to prevent the growth of metastases and to treat metastatic disease once this has become clinically apparent. Current regimens are based on empirical drug combinations, and few clinical trials have been conducted because of the rarity of this tumour. Chemosensitivity testing offers a way of testing a large number of agents against tumours. The ATP-based chemosensitivity assay (ATP-TCA) has already helped to design new regimens for melanoma and breast and ovarian cancer. Primary retinoblastoma tumour material was obtained from 10 eyes, 7 of which contained sufficient viable cells for ATP TCA. The results show very high sensitivity to single agents, particularly cisplatin, doxorubicin and vinca aLkaloids. Of the anti-metabolites tested, 5-FU is relatively disappointing (although still active), and gemcitabine shows considerable activity consistent with a cytotoxic effect. The shape of the inhibition curves is interesting. There is a plateau effect with the topoisomerase inhibitors and vinblastine, which is not present with the cisplatin. One tumour was much more resistant than the others tested, particularly to vinblastine but also to the topoisomerase inhibitors, which failed to achieve complete kill at any concentration tested, consistent with a multidrug resistance phenotype. Of the combinations (VAC and VEC), the VAC regimen looks marginally more active in the more resistant of the two cases tested to date. These data confirm that retinoblastoma is a rapidly growing malignancy that is very susceptible to cytotoxic drugs of all types. Chemosensitivity testing provides a practical method of testing new regimens before clinical trials in retinoblastoma patients. PMID- 12528801 TI - Chemosensitivity testing in malignant melanoma. AB - The prognosis of patients with metastatic melanoma remains poor. In patients with distant metastases only low response rates between 10% and 15% have been achieved by the most effective cytostatics in single-agent therapy leading to a mean 5 year survival rate of less than 5%. More aggressive treatment regimens using multidrug chemotherapy yielded response rates of up to 40% but failed to show a significant benefit in overall survival compared to single-agent therapy. However, complete remissions of metastatic lesions after multidrug cytostatic regimens have been reported in some cases of melanoma patients. To evaluate an in vitro test system providing information on the drug sensitivity profile of melanoma cells, we examined tumor tissue specimens from 31 metastatic melanoma patients with an ATP-based chemosensitivity assay (ATP-TCA) testing eight anticancer drugs alone or in different combinations. Chemosensitivity was assessed using a luciferin-luciferase- based luminescence assay providing individual chemosensitivity indices for each test drug. We found a heterogeneous chemosensitivity in the melanoma tissue samples tested. The highest sensitivity was detected for the combination of treosulfan and gemcitabine, with 76% of the tissue samples revealing high sensitivity and 10% resistance, followed by the combination of paclitaxel and doxorubicine (66%/0%), gemcitabine and cisplatin (55%/21%),and paclitaxel and cisplatin (46%/8%). Our data indicate that the ATP TCA can be used to select patients who might benefit from an individually adapted cytostatic therapy. On the basis of these results a multicenter trial has recently been initiated to evaluate the feasibility and predictive value of an ATP-TCA directed chemotherapy in metastatic melanoma patients. PMID- 12528802 TI - Human melanoma: drug resistance. AB - Advanced malignant melanoma has a poor prognosis since chemotherapy is mostly ineffective because, in part, of the intrinsic and/or extrinsic resistance of melanoma cells to systemic treatment with antineoplastic agents. The reasons for the chemoresistant phenotype are currently unknown. The relevance of well analyzed drug resistance mechanisms in melanoma such as intracellular and extracellular transport, drug resistance by induction of certain enzyme systems, and altered drug-target interaction is reviewed. It has been shown that most anticancer drugs kill susceptible cells through induction of apoptosis. Therefore, the significance of apoptotic deficiency caused by alteration in the apoptotic pathway is discussed in relation to specific molecules and apoptotic mechanisms like death-receptors, the Bcl-2 family, and the Hsp family of proteins. The complexity of the molecular variants involved in signal transduction along apoptotic pathways suggests that the cell may possess a variety of possibilities for regulating apoptosis and generating apoptosis deficiency. Thus apoptosis and apoptosis deficiency should be analyzed to understand the mechanisms of melanoma resistance. PMID- 12528803 TI - Cisplatin, doxorubicin and paclitaxel induce mdr1 gene transcription in ovarian cancer cell lines. AB - The clinical observation of the multidrug resistance (MDR) phenotype is often associated with overexpression of the mdrl gene, in particular with respect to ovarian cancer. However, until now the mdrl-inducing potential of commonly used antineoplastics has been only incompletely explored. We performed short-term cultures of six ovarian cancer cell lines (MZOV4, EF027, SKOV3, OAW42, OTN14, MZOV20) exposed to either blank medium or cisplatin, doxorubicin or paclitaxel at concentrations related to the clinically achievable plasma peak concentration. A highly specific quantitative real-time RT-PCR was used to detect the Mdr1 transcripts. Mdrl mRNA contents were calibrated in relation to coamplified GAPDH mRNA. Mdrl mRNA was detectable in each cell line. In 13 out of 18 assays (72%) the specific anticancer drug being tested induced mdr1 transcription. No decrease in mdr1 mRNA concentration was observed. Our data suggest that mdr1 induction by antineoplastics is one of the reasons for failure of ovarian cancer therapy but may vary individually. PMID- 12528804 TI - Chemosensitivity testing as an aid to anti-cancer drug and regimen development. AB - The ATP-based chemosensitivity assay has proved particularly useful for the evaluation of new anti-cancer agents and combinations. The majority of our publications in this area have concentrated on topoisomerase inhibitors. Comparison of mitoxantrone with doxorubicin convinced us that these two agents were not completely cross-resistant and led to the design of the mitoxantrone + paclitaxel regimen which is now in clinical practice. Re-assessment of treosulfan in uveal melanoma led to the design of a new regimen combining this alkylating agent with gemcitabine, again with rapid introduction of this combination to clinical practice. The assay has recently been used to examine the concentration activity curve to determine which tumours might benefit from liposomal preparations capable of delivering 4-16 times the standard dose without cardiotoxicity. Assay-directed use of Caelyx is producing encouraging results, and we are now examining this drug in combination with others. We recently showed that XR5000, a combined inhibitor of topoisomerase I and II, was effective against melanoma as well as ovarian cancer, but at concentrations which were unlikely to be achieved in patients. These data confirm our suggestion that use of the assay could reduce the time to introduction of new anti-cancer drugs and the cost of this process. PMID- 12528805 TI - Assay-assisted treatment selection for women with breast or ovarian cancer. AB - Although women suffering from advanced cancer of the breast or ovary are unlikely to be cured, several active agents are available that can prolong their lives. The use of these agents is based on demonstrated benefit in large randomized clinical trials, and the clinical activity of these chemotherapy regimens is initially high, with 60%-70% of patients responding. Unfortunately, their benefit in the second-line setting is often limited, with less than 30% of patients showing significant disease response. Thus some 70% of patients may undergo ineffective treatment during the course of their disease, while still suffering from significant chemotherapy-related toxicity. Having some foreknowledge of a given agent's expected result before its administration would therefore benefit the individual patient. In vitro drug response testing, first developed to assist in the selection of antibiotics for patients with bacterial infections, has recently been demonstrated to accurately predict how cancer patients will respond to chemotherapy. This review discusses the historical development of in vitro testing for cancer patients, some of the pitfalls encountered, and offers an assessment of their current utility. Results of various clinical trials that evaluated correlations between in vitro tumor response and clinical outcomes are described. These data suggest that in vitro drug response assays can accurately predict drug resistance and can identify patients who are more or less likely to benefit from a given agent. PMID- 12528806 TI - Chemosensitivity testing in gynecologic oncology--dream or reality? AB - Cell culture and animal models have played an essential role in the research of new principles of therapy. Many methods for the individualized testing of therapy sensitivity and resistance have been developed, for example, the clonogenic assay. Presently, the ATP-TCA is commercially available as a testing kit. This review gives an overview of the tumor samples that were tested in the oncologic laboratory in the Department of Obstetrics and Gynecology, Munich Grosshadern between 1993 and 2001. All target parameters show a clear trend in favor of sequential, dose-intensified Epirubicin/Paclitaxel therapy. If this trend remains valid for the total number of patients, a significant impact of this new principle of therapy can be expected. By individualized planning of therapy with ATP-TCA testing, therapy in the individual patient could already be performed by the examination of sensitivity in the preoperative biopsy specimen. PMID- 12528807 TI - Treosulfan in the treatment of metastatic melanoma: from chemosensitivity testing to clinical trials. AB - The therapy of metastatic malignant melanoma is limited by poor responses and short overall survival. Thus it remains important to identify and test potential new drugs in this disease. To examine the effects of the bifunctional alkylating cytostatic treosulfan, an in vitro microplate ATP bioluminescence assay (ATP-TCA) was used. Five highly chemoresistant melanoma cell lines and melanoma cells freshly isolated from metastases surgically resected from stage IV melanoma patients (n = 10) were incubated with treosulfan. Three cell lines and eight of ten tested tumor cells isolated from melanoma metastases showed tumor growth inhibition after incubation with treosulfan. Therefore, 14 patients with rapidly progressing stage IV malignant melanoma who were pretreated with at least one standard chemotherapy regimen received treosulfan. In this population of patients with highly refractory advanced melanoma one complete remission (7.1%), two partial remissions (14.3%), and three cases of stable disease (21.4%) were observed. Median time to progression and median overall survival for all patients measured from the beginning of treosulfan treatment were 5 months [95% confidence interval (CI) 1.98-2.57 months] and 9 months (95% CI 3.92-8.69 months), respectively. On the basis of these data a multicenter phase II trial was initiated. A total of 31 patients with stage IV melanoma were included and treated second-line with 8 g/m2 i.v. treosulfan. From this group 26 patients were evaluable. No objective remission (CR, PR) was observed, 5 of 26 patients (19%) had stable disease, and 21 patients had progressive disease. Median overall survival was 6.5 months (95% CI 3.1-10 months). Toxicity of treosulfan was moderate. Patients with treosulfan-sensitive melanoma metastases showed better response rates and prolonged survival compared with patients who were not tested before treosulfan treatment. We therefore suggest further studies with first-line treosulfane alone or in combination with gemcitabine or cytosine arabinoside together with pretherapeutic chemosensitivity testing that may help to select patients who might benefit from specific chemotherapy. PMID- 12528808 TI - Chemosensitivity testing and test-directed chemotherapy in human pancreatic cancer. AB - Human pancreatic cancer is a devastating disease with poor prognosis. In many cases it is diagnosed at stages in which a complete resection is not possible. However, even after complete resection most tumors recur. Therefore, several chemotherapeutic strategies have been developed, so far, with little impact on the clinical outcome. Because one of the hallmarks of human pancreatic cancer is its general resistance to chemotherapeutic agents, it seems important to develop strategies to individualize chemotherapy and to render cells more sensitive to chemotherapeutic agents. In this summary we describe our methods of in vitro chemosensitivity testing using the human tumor colony-forming assay for pancreatic cancer in comparison with other solid tumors and describe how the in vitro results influence chemotherapy. Furthermore, we point out new developments of mRNA quantitation of chemoresistance target enzymes based on real-time PCR, which may help in the future to individualize chemotherapy of pancreatic cancer. Finally, we present results of studies of cyclin D1 inhibition. Suppression of cyclin D1 by cyclin D1 antisense mRNA expression was associated with growth inhibition and an increase in chemosensitivity to fluoropyrimidines and platinum compounds. Because human pancreatic cancers are relatively chemoresistant and material for chemosensitivity testing with the human tumor colony-forming assay (HTCA) is in most cases difficult to obtain, future investigations should aim at the development of methods requiring only very small samples to analyzemarkers of chemosensitivity. Our results further suggest that chemotherapy in combination with strategies to increase chemosensitivity may be a reasonable regimen for the treatment of human pancreatic cancer in the future. PMID- 12528809 TI - Clinical significance of cellular drug resistance in childhood leukemia. AB - Cellular drug resistance is an important determinant of the response to chemotherapy, and its precise measurement may have clinical relevance. Potential applications are: prognostic factor for risk-group stratification, tailored chemotherapy for subgroups or individual patients with a specific cellular drug resistance profile, determination of cross-resistance patterns, study of drug interactions, study of resistance modulation or circumvention, selection of patients for phase II studies and screening for the cytotoxicity of novel compounds. The colorimetric 4-day MTT assay is a frequently used method. However, a distinction between malignant and non-malignant cells cannot be made, which should be taken into account. In the case of a relatively high percentage of contaminating non-malignant cells, the differential staining cytotoxicity (DiSC) assay can be used. The MTT assay's technical success percentage is about 80% for fresh ALL and AML samples. For methotrexate (MTX) a different assay must be used, such as the thymidylate synthase inhibition assay (TSIA). The MTT assay measures the number of living cells that survived drug exposure. Therefore, the effect of many if not most drugs to induce leukemia cell death by apoptosis is also included. This review mainly summarizes the data on cellular drug resistance in childhood leukemia, as obtained by the MTT assay and TSIA, in our laboratory in Amsterdam. These data clearly demonstrate the significant relation between in vitro cellular drug resistance and clinical and cell biological features and short- and long-term clinical outcome in childhood leukemia. In conclusion, cellular drug resistance testing provides clinically relevant information that can be available within 1 week and can be performed successfully in the vast majority of leukemia samples. The data are more and more being used and being considered for use in clinical trials in leukemia. PMID- 12528810 TI - ATP chemosensitivity testing in ovarian and breast cancer: early clinical trials. AB - After disappointing results achieved with older chemosensitivity tests such as the human tumor clonogenic assay (HTCA) during the 1980s, the last decade has seen a renaissance of the concept of individualized chemotherapy in oncology, markedly stimulated by the development of newer nonclonogenic assays. These methods appear to be able to overcome major technical limitations associated with older assays, now allowing for successful testing of most of the tumor specimens submitted. Currently, the ATP-based tumor chemosensitivity assay (ATP-TCA) can be regarded as the most sophisticated assay to investigate both solid samples and effusions derived from patients with various organ tumors. During the last 5 years, the ATP-TCA has been used successfully to screen for novel drug combinations for further clinical use in both ovarian and breast cancer such as mitoxantrone plus paclitaxel (NT) and treosulfan plus gemcitabine (TG), respectively. Clinical trials that have been set up in heavily pretreated patients with recurrent ovarian or breast cancer have convincingly confirmed the high activity of these combinations previously demonstrated in preclinical investigations using the ATP-TCA. In a recent phase II trial performed in 59 patients with relapsed ovarian carcinoma, ATP-TCA-directed therapy was able to triple the response rate and to double the survival time, compared with published empirical chemotherapy regimes. Preliminary results with ATP-TCA-directed therapy in breast cancer also evidenced promising response rates. These results have been confirmed by additional prospective clinical trials using other types of modern nonclonogenic assays. A phase III trial that is now actively recruiting patients with platinum-refractory ovarian cancer to verify the promising phase II studies will prove the further value of the ATP-TCA as a predictor applicable in routine clinical oncology. PMID- 12528811 TI - Chemosensitivity testing--present and future in Japan. AB - Radical surgery with extended lymph node dissection is the first and only curative treatment of gastrointestinal cancer. Although the combined cancer chemotherapy has achieved 30%-50% response rates, a controversy still remains over the significance of the adjuvant cancer chemotherapy after surgery. To break through this limitation, we have introduced the chemosensitivity test to evaluate the appropriate adjuvant cancer chemotherapy for advanced gastrointestinal cancer. Our plural studies indicated that the chemosensitivity test would be useful in evaluating the appropriated adjuvant chemotherapy by increasing survival in the sensitive group. Recently, the molecular targets have been clarified for the conventionally available antitumor agents, e.g., thymidylate synthetase for 5-fluorouracil, ATP-binding cassette transporters for anthracyclines, glutathione-related detoxification for platins, and topoisomerase I for CPT-11, which will be applied for dinical use in evaluating the appropriate cancer chemotherapy. The chemosensitivity test is approved as "advanced clinical medicine" by the Japanese Ministry of Health, Welfare, and Labor in five institutes at present. Because complete dissection and chemosensitivity test guided adjuvant chemotherapy will result in a survival benefit for patients with advanced gastrointestinal cancer, this test should be approved as "social insurance" for further wide clinical application. PMID- 12528812 TI - Adult blood lead epidemiology and surveillance--United States, 1998-2001. AB - PROBLEM/CONDITION: Elevated blood lead levels (BLLs) in adults can damage the cardiovascular, central nervous, reproductive, hematologic, and renal systems. The majority of cases are workplace-related. U.S. Department of Health and Human Services recommends that BLLs among all adults be reduced to < 25 microg/dL. The highest BLL acceptable by standards of the U.S. Occupational Safety and Health Administration is 40 microg/dL. The mean BLL of adults in the United States is < 3 microg/dL. REPORTING PERIOD: This report covers cases of adults (aged > or = 16 years) with BLLs > or = 25 microg/dL, as reported by 25 states during 1998-2001. DESCRIPTION OF SYSTEM: Since 1987, CDC has sponsored the state-based Adult Blood Lead Epidemiology and Surveillance (ABLES) program to track cases of elevated BLLs and provide intervention consultation and other assistance. Overall ABLES program data were last published in 1999 for the years 1994-1997. This report provides an update with data from 25 states reporting for > or = 2 years during 1998-2001. During that period, the ABLES program funded surveillance in 21 states - Alabama, Arizona, Connecticut, Iowa, Maryland, Massachusetts, Michigan, Minnesota, New Jersey, New York, North Carolina, Ohio, Oklahoma, Oregon, Pennsylvania, Rhode Island, South Carolina, Texas, Washington, Wisconsin, and Wyoming. Four additional states - California, Nebraska, New Hampshire, and Utah contributed data without CDC funding. RESULTS: During 1998-2001, the overall program's annual mean state prevalence rate for adults with BLLs > or = 25 microg/dL was 13.4/100,000 employed adults. This compares with 15.2/100,000 for 1994-1997. Yearly rates were 13.8 (1998), 12.9 (1999), 14.3 (2000), and 12.5 (2001). For adults with BLLs > or = 40 microg/dL, the overall program's annual mean state prevalence rare during 1998-2001 was 2.9/ 100,000 employed adults. This compares with 3.9/100,000 for 1994-1997. Yearly rates were 3.3 (1998), 2.5 (1999), 2.9 (2000), and 2.8 (2001). INTERPRETATION: Although certain limitations exist, the overall ABLES data indicate a declining trend in elevated BLLs among employed adults. PUBLIC HEALTH ACTIONS: ABLES-funded states increased from 21 to 35 in 2002, and more detailed reporting requirements were put into effect. These, and other improvements, will enable the ABLES program to work more effectively toward its 2010 target of eliminating all cases of BLLs > or = 25 microg/dL in adults caused by workplace exposures. PMID- 12528813 TI - The neuronal ceroid-lipofuscinoses. AB - The neuronal ceroid-lipofuscinoses (NCLs) collectively constitute the most common group of neurodegenerative diseases in childhood and usually show an autosomal recessive mode of inheritance. Despite varying ages of onset and clinical course characterized in most instances by progressive mental and motor deterioration, blindness, epileptic seizures, and premature death, all forms of NCL show unifying histopathological features. There is accumulation of autofluorescent, periodic acid-Schiff-, and Sudan black B-positive granules that are resistant to lipid solvents in the cytoplasm of most nerve cells and. to a lesser degree, of many other cell types. The storage process is associated with progressive and selective neuronal loss and gliosis with secondary white matter lesions. The ultrastructure of the storage deposits varies between different forms of NCL and, along with the age of onset, has provided the basis for the traditional classification of NCLs. Recent molecular genetic findings have established that defects in at least 7 different genes underlie the various forms of NCL. The purpose of this paper is to provide an overview of the NCLs, review recent molecular genetic and biochemical findings, and discuss their impact on our views on the classification and pathogenesis of these devastating brain disorders. PMID- 12528814 TI - Transglutaminase cross-links in intranuclear inclusions in Huntington disease. AB - Cortical and striatal perinuclear cytoplasmic aggregates and intranuclear inclusions of mutant huntingtin are neuropathological hallmarks of Huntington disease (HD). Although the mechanisms involved in the formation of these aggregates are unclear, a recent hypothesis implicates cross-linking of mutant huntingtin protein into aggregates by transglutaminase. This study explores the hypothesis that transglutaminase catalyzes cross-linking of huntingtin into intranuclear inclusions. Using immunofluorescence and confocal microscopy we demonstrate 99% colocalization of transglutaminase-catalyzed epsilon-(gamma glutamyl) lysine covalent cross-links with nuclear aggregates of huntingtin protein in the frontal cortex of postmortem HD brain tissue. Furthermore, the transglutaminase 2 isoform colocalizes with both huntingtin protein and epsilon (gamma-glutamyl) lysine covalent cross-links in HD intranuclear inclusions. Transient transfection of N-terminally truncated huntingtin with an expanded glutamine domain (htt-N63-148Q-myc) with and without and transglutaminase 2 into HEK 293T cells resulted in an increase in cross-linked huntingtin in the insoluble formic acid-treated pellet in comparison to transfection of N terminally truncated huntingtin with normal length glutamine domain (htt-N63-18Q myc). Transfection with both htt-N63-148Q-myc and transglutaminase 2 resulted in high molecular weight huntingtin in the insoluble fraction. These data support the hypothesis that transglutaminase catalyzed cross-linking of mutant huntingtin is involved in the formation and/or stabilization of huntingtin protein aggregates in HD. Based on these and other studies, modulation of transglutaminase activity could be explored as a treatment for HD. PMID- 12528815 TI - Preferential loss of myelin-associated glycoprotein reflects hypoxia-like white matter damage in stroke and inflammatory brain diseases. AB - Destruction of myelin and oligodendrocytes leading to the formation of large demyelinated plaques is the hallmark of multiple sclerosis (MS) pathology. In a subset of MS patients termed pattern III, actively demyelinating lesions show preferential loss of myelin-associated glycoprotein (MAG) and apoptotic-like oligodendrocyte destruction, whereas other myelin proteins remain well preserved. MAG is located in the most distal periaxonal oligodendrocyte processes and primary "dying back" oligodendrogliopathy may be the initial step of myelin degeneration in pattern III lesions. In the present study, various human white matter pathologies, including acute and chronic white matter stroke, virus encephalitis, metabolic encephalopathy, and MS were studied. In addition to a subset of MS cases, a similar pattern of demyelination was found in some cases of virus encephalitis as well as in all lesions of acute white matter stroke. Brain white matter lesions presenting with MAG loss and apoptotic-like oligodendrocyte destruction, irrespective of their primary disease cause, revealed a prominent nuclear expression of hypoxia inducible factor-1alpha in various cell types, including oligodendrocytes. Our data suggest that a hypoxia-like tissue injury may play a pathogenetic role in a subset of inflammatory demyelinating brain lesions. PMID- 12528816 TI - Characterization of an O-glycosylated plaque-associated protein from Alzheimer disease brain. AB - In this work we characterized a 90-kDa glycoprotein from Alzheimer disease (9OAzgp) brain extracts that is recognized by the GalNAc-specific lectin from Amaranthus leucocarpus (ALL), as determined through Western blot. The 90Azgp was purified by electro-elution, and its amino acid sequence determined from peptides obtained after trypsin digestion through MALDI-TOF (Matrix-assisted laser desorption ionization-time of flight), and compared with the relative values obtained from the NCBInr (Swiss-Prot 10/01/2001) database. The 90Azgp showed 32% and 42% homology with the KIAA0310 protein from human brain and the human gastric mucin, respectively. Presence of O-glycosidically linked glycans in the proteins recognized by ALL was confirmed by inhibition of the lectin-glycoprotein interaction through hapten-inhibition assays and also by elimination of the O glycosidically linked glycans after treatment with O-glycanase from Diplococcus pneumoniae. Electron transmission microscopy confirmed that the receptor recognized by the lectin is processed in the Golgi apparatus of AD neurons. Although the specific role of this glycoprotein has not been identified, considering that the presence of this lectin receptor co-localized with neuritic plaques and in AD sprouting neurons, it could suggest that the O-glycosyl-protein identified by the A. leucocarpus lectin participates in the pathogenesis of neurodegenerative diseases. PMID- 12528817 TI - Neuregulin-1 and erbB4 immunoreactivity is associated with neuritic plaques in Alzheimer disease brain and in a transgenic model of Alzheimer disease. AB - Neuregulin-1 (NRG-1) regulates developmental neuronal survival and synaptogenesis, astrocytic differentiation, and microglial activation. Given these NRG-1 actions, we hypothesized that the synaptic loss, gliosis, inflammation, and neuronal death occurring in Alzheimer disease (AD) is associated with altered expression of NRG-1 and its receptors (the erbB2, erbB3, and erbB4 membrane tyrosine kinases). We examined the expression and distribution of NRG-1 and the erbB kinases in the hippocampus of AD patients and cognitively normal controls and in transgenic mice that coexpress AD-associated mutations of the beta amyloid precursor protein (APP(K670N,M671L)) and presenilin-1 (PS1(M146L)). In the hippocampi of both control humans and wild type mice, NRG-1 and the 3 erbB receptors are expressed in distinct cellular compartments of hippocampal neurons. All 4 molecules are associated with neuronal cell bodies, but only NRG-1, erbB2, and erbB4 are present in synapse-rich regions. In AD and in the doubly transgenic mouse, erbB4 is expressed by reactive astrocytes and microglia surrounding neuritic plaques. In AD brains, microglia and, to a lesser extent, dystrophic neurites, also upregulate NRG-1 in neuritic plaques, suggesting that autocrine and/or paracrine interactions regulate NRG-1 action within these lesions. NRG-1 and erbB4, as well as erbB2, are similarly associated with neuritic plaques in the doubly transgenic mice. Thus, in AD the hippocampal distribution of NRG-1 and erbB4 is altered. The similarities between the alterations in the expression of NRG-1 and its receptors in human AD and in APP(K670N;M671L)/PS1(M146L) mutant mice suggests that this animal model may be very informative in deciphering the potential role of these molecules in AD. PMID- 12528818 TI - Stereologic evidence for persistence of viable neurons in layer II of the entorhinal cortex and the CA1 field in Alzheimer disease. AB - The entorhinal cortex and hippocampus are the first cortical regions to be affected by the degenerative cellular process that leads to Alzheimer disease (AD) and display a limited degree of neuronal alterations in normal aging. Several quantitative studies have reported a substantial loss of neurons in these regions and a parallel increase in the number of neurofibrillary tangles (NFTs). However, accurate quantitative data on the dynamics of NFT formation are lacking. Here, we performed a stereologic assessment of the proportions of intracellular and extracellular (ghost) NFTs (iNFTs and eNFTs, respectively) and unaffected neurons in layer II of the entorhinal cortex and in the pyramidal cell layer of the CA1 field of the hippocampus in elderly control cases compared to cases with varying degrees of cognitive dysfunction. The data revealed differential rates of formation of iNFTs and eNFTs between the 2 regions and confirmed the presence of a severe disease-associated, but not age-related, neuronal loss. They also revealed that large numbers of neurons may persist either unaffected or in a transitional stage of NFT formation until the late stages of AD progression. These neurons with viability potential constitute 73% of the total numbers of profiles in layer II of the entorhinal cortex and 77% in the CA1 field in cases with a Clinical Dementia Rating score of 3. Whereas it is not possible in the present study to assess how functional such neurons with altered physiology might be, it is nonetheless likely that these transitional neurons open new options for potential therapeutic interventions aimed at protecting neurons vulnerable to neurofibrillary degeneration. PMID- 12528819 TI - Expression patterns of retinoblastoma protein in Parkinson disease. AB - Cellular mechanisms implicated in Parkinson disease (PD) include oxidative stress, inflammatory response, excess dopamine, DNA damage, and loss of trophic support. These stimuli have been observed to induce changes in cell cycle proteins in several cell types. One of the key regulators of cell cycle progression is the retinoblastoma protein (pRb); therefore, we assessed the staining for pRb and its inactive hyperphosphorylated isoform, ppRb, in autopsy tissue from patients with PD. In PD we found abundant pRb staining in neuronal cytoplasm of the substantia nigra, mid-frontal cortex, and hippocampus by immunohistochemistry. In controls, pRb weakly stained nucleoli of neurons in the substantia nigra and exhibited no detectable staining in mid-frontal cortex and hippocampus. Staining for ppRb resulted in a shift from weak cytoplasmic staining in neurons from control cases to strong nuclear staining in PD cases, especially within the substantia nigra, mid-frontal cortex, and hippocampus. In the substantia nigra, ppRb also co-localized to Lewy bodies, which are a pathologic feature of PD. Lewy bodies are also found in diffuse Lewy body disease (DLBD) that do not consistently exhibit changes in pRb or ppRb. These results indicate that there are changes in pRb and its inactive phospho-isoform in neurons responding to neurodegenerative stimuli associated with PD. PMID- 12528820 TI - Axotomized motoneurons can be rescued from cell death by peripheral nerve grafts: the effect of donor age. AB - Injury to neonatal nerves, unlike adult nerves, results in poor regeneration and extensive motoneuron death. We examined whether exposure to a more mature nerve environment could rescue axotomized motoneurons following neonatal injury. The sciatic nerve in 1 hindlimb of 3-day-old (P3) rats was transected and the cut end sutured to a nerve graft taken from donor rats, which ranged between P3 and P21. The extent of motoneuron survival and axon regeneration was established 7 days later. Since integrins play an important role in regeneration, we also examined the effect of manipulating integrin binding in nerve grafts. Following axotomy at P3 and implantation of nerve grafts from 3-day-old rats, approximately 38% of motoneurons survived. In contrast, grafts from rats aged 5 days and older resulted in an improvement in regeneration, and over 70% of motoneurons survived. This survival-promoting effect of P5 grafts was prevented by blocking beta1 integrins. In contrast, increasing beta1-integrin levels in grafts from P3 rats dramatically increased motoneuron survival. Thus, following neonatal nerve injury, exposure to a more mature nerve environment significantly increases motoneuron survival, an effect that is dependent upon beta1-integrin signaling. Therefore, pharmacological upregulation of beta1-integrins may significantly improve the outcome of neonatal nerve injuries. PMID- 12528821 TI - PARP expression is increased in astrocytes but decreased in motor neurons in the spinal cord of sporadic ALS patients. AB - The evidence for increased oxidative stress and DNA damage in amyotrophic lateral sclerosis (ALS) prompted studies to determine if the expression of poly(ADP ribose) polymerase (PARP) is increased in ALS. Using Western analyses of postmortem tissue, we demonstrated that PARP-immunoreactivity (PARP-IR) was increased 3-fold in spinal cord tissues of sporadic ALS (sALS) patients compared with non-neurological disease controls. Despite the increased PARP-IR, PARP mRNA expression was not increased significantly. Immunohistochemical analyses revealed PARP-IR was increased in both white and gray matter of sALS spinal cord. While PARP-IR was predominantly seen in astrocytes, large motor neurons displayed reduced staining compared with controls. This result contrasts sharply to the staining of Alzheimer and MPTP-induced Parkinson diseased tissue, where poly(ADP ribose) (PAR)-IR was seen mostly in neurons, with little astrocytic staining. PARP-IR was increased in the pellet fraction of sALS homogenates compared with control homogenates, representing potential PARP binding to chromatin or membranes and suggesting a possible mechanism of PARP stabilization. The present results demonstrate glial alterations in sALS spinal cord tissue and support the role of glial alterations in sALS pathogenesis. Additionally, these results demonstrate differences in sALS spinal motor neurons and astrocytes compared to brain neurons and astrocytes in Alzheimer disease and MPTP-induced Parkinson disease despite the presence of markers for oxidative stress in all 3 diseases. PMID- 12528822 TI - Hypertensive cerebral hemorrhage. Demonstration of the source of bleeding. PMID- 12528823 TI - Associations between air emissions from sour gas processing plants and indices of cow retainment and survival in dairy herds in Alberta. AB - This paper describes the results of an investigation into the effects of air emissions from sour gas processing plants on indices of retainment or survival of adult female dairy cattle on farms in Alberta; namely, the productive lifespan of individual animals, and annual herd-level risks for culling and mortality. Using a geographical information system, 2 dispersion models--1 simple and 1 complex- were used to assess historical exposures to sour gas emissions at 1382 dairy farm sites from 1985 through to 1994. Multivariable survival models, adjusting for the dependence of survival responses within a herd over time, as well as potential confounding variables, were utilized to determine associations between sour gas exposure estimates and the time from the first calving date to either death or culling of 150210 dairy cows. Generalized linear models were used to model the relationship between herd-level risks for culling and mortality and levels of sour gas exposure. No significant (P < 0.05) associations were found with the time to culling (n = 70052). However, both dispersion model exposure estimates were significantly associated (P < 0.05) with a decreased hazard for mortality; that is, in cases where cattle had died on-farm (n = 8743). There were no significant associations (P > 0.05) between herd culling risks and the 2 dispersion model exposure estimates. There was no measurable impact of plant emissions on the annual herd risk of mortality. PMID- 12528824 TI - Mechanical transmission of porcine reproductive and respiratory syndrome virus throughout a coordinated sequence of events during warm weather. AB - Mechanical transmission of porcine reproductive and respiratory syndrome virus (PRRSV) throughout a coordinated sequence of events that replicated common farm worker behavior during warm weather (10 degrees C to 16 degrees C) was assessed using a field-based model. The model involved fomites (boots and containers), vehicle sanitation, transport, and personnel movement. In a previous study, the model successfully demonstrated mechanical transmission of PRRSV in 8 out of 10 replicates during cold weather. A field strain of PRRSV was inoculated into carriers consisting of soil samples, which were adhered to the undercarriage of a vehicle. The vehicle was driven approximately 50 km to a commercial truck washing facility where the driver's boots contacted the carriers during washing, introducing the virus to the vehicle interior. The vehicle was then driven 50 km to a simulated farm site, and the driver's boots mechanically spread virus into the farm anteroom. Types of containers frequently employed in swine farms contacted drippings from the footwear on the anteroom floor. The truck wash floor, vehicle cab floor mats, boot soles, anteroom floor, and the ventral surface of containers were sampled to track the virus throughout the model. Ten replicates were conducted, along with sham-inoculated controls, and control replicates. In 2 replicates, infectious PRRSV was detected on the anteroom floor and in 1 replicate, infectious PRRSV was detected on the surface of the container by swine bioassay. All sham-inoculated controls and protocol controls were negative. These results indicate that mechanical transmission of PRRSV throughout a coordinated sequence of events in warm weather can occur, but in contrast to data from studies conducted during cold weather, it appears to be a relatively infrequent event. PMID- 12528825 TI - Diagnostic decision rule for support in clinical assessment of the need for surgical intervention in horses with acute abdominal pain. AB - A prospective survey of horses with colic referred to a university hospital was undertaken to elaborate on a simple clinical decision support system capable of predicting whether or not horses require surgical intervention. Cases were classified as requiring surgical intervention or not on the basis of intraoperative findings or necropsy reports. Logistic regression analysis was applied to identify predictors with the strongest association with treatment needed. The classification and regression tree (CART) methodology was used to combine the variables in a simple classification system. The performance of the elaborated algorithms, as diagnostic instruments, was recorded as test sensitivity and specificity. The CART method generated 5 different classification trees with a similar basic structure consisting of: degree of pain, peritoneal fluid colour, and rectal temperature. The tree, constructed at a prevalence of 15% surgical cases, appeared to be the best proposal made by CART. In this classification tree, further discrimination of cases was obtained by including the findings of rectal examination and packed cell volume. When regarded as a test system, the sensitivity and specificity was 52% and 95%, respectively, corresponding to positive and negative predictive values of 68% and 91%. The variables examined in the present study did not provide a safe clinical decision rule. The classification tree constructed at 15% surgical cases was considered feasible, the proportion of horses incorrectly predicted to be without need of immediate surgery (false negatives) was small, whereas the proportion of horses incorrectly predicted to be in need of immediate surgery (false positives) was large. Some of the false positive horses were amenable to surgical treatment, although these cases did not conform to the strict definition of a surgical case. A less rigorous definition of a surgical case than that used in the present study would lower the percentage of false positives. PMID- 12528826 TI - Urethral pressure profile and hemodynamic effects of phenoxybenzamine and prazosin in non-sedated male beagle dogs. AB - Prazosin is a readily available alpha-adrenergic antagonist that may be useful in the management of functional urethral obstruction in companion animals. This study used urethral pressure profilometry to evaluate the urethral effects of prazosin and phenoxybenzamine in healthy, non-sedated, male Beagle dogs. Heart rate, indirect systolic, diastolic and mean arterial blood pressures were measured, and saline perfusion urethral pressure profilometry was performed at 0, 10, 20, and 40 min following intravenous administration of prazosin (0.025 mg/kg), phenoxybenzamine (0.2 mg/kg), or placebo. Maximal urethral pressure, maximal urethral closure pressure, post peak nadir, and all blood pressure parameters decreased significantly at nearly all treatment intervals following administration of prazosin compared with placebo. Less consistently significant reductions were observed following phenoxybenzamine administration. Maximal decreases in urethral pressure parameters were observed 20 min following the injection of prazosin; maximal blood pressure decreases were evident by 10 min postinjection. In this non-sedated dog model, urethral pressure profilometry was a sensitive method of detecting urethral effects of alpha antagonists. Repeatable reductions in urethral pressure measurements were observed, with prazosin effecting more consistently significant changes than phenoxybenzamine. Significant decreases in systolic, diastolic, and mean arterial blood pressures were seen with prazosin, but not phenoxybenzamine or placebo. Further study of selective alpha-1 antagonists in dogs is needed to determine appropriate oral dosing protocols that will produce maximal urethral effects with minimal hemodynamic effects, and to demonstrate clinical efficacy in dogs with functional urethral obstruction. PMID- 12528828 TI - Adherence of bovine viral diarrhea virus to bovine oocytes and embryos with a hardened zona pellucida cultured in vitro. AB - The purpose of this study was to investigate the adherence of bovine viral diarrhea virus (BVDV) to bovine mature, or immature, cumulus-free oocytes and to in vitro fertilized embryos, maintained in vitro in a ligated bovine oviduct to allow for the hardening of the zona pellucida. Incubation of the oocytes and embryos in the oviduct for 5 h caused hardening of the zona pellucida as measured by resistance to pronase digestion (which increased from approximately 3 min to 7 h; P > 0.001). However, there was no difference between the number of infected oocytes and embryos (n = 965 in 193 samples) following experimental exposure to BVDV regardless of whether or not they were previously incubated in the oviduct (P > 0.05). It was concluded that the modification of the proteolytic resistance properties of the zona pellucida during in vitro oviductal incubation did not influence the adherence of BVDV to zona pellucida of oocytes or in vitro fertilized embryos. PMID- 12528827 TI - Characterization of integron mediated antimicrobial resistance in Salmonella isolated from diseased swine. AB - Forty-two Salmonella isolates obtained from diseased swine were genetically characterized for the presence of specific antimicrobial resistance mechanisms. Twenty of these isolates were characterized as S. Typhimurium DT104 strains. Pulsed-field gel electrophoresis was used to determine genetic relatedness and revealed 20 distinct genetic patterns among the 42 isolates. However, all DT104 isolates fell within 2 closely related genetic clusters. Other Salmonella isolates were genetically grouped together according to serotype. All DT104 isolates displayed the penta-resistance phenotype to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline. Resistance to sulfamethoxazole, tetracycline, streptomycin, kanamycin, and ampicillin was most common among the non-DT104 Salmonella isolates. All DT104 strains contained 2 chromosomal integrons of 1000 and 1200 base pairs. The DNA sequencing revealed that the 2 integrons contained genes encoding a resistance to streptomycin and ampicillin, respectively. None of the non-DT104 strains showed the same pattern, although several strains possessed integrons of 1000 base pairs or larger. However, the majority of non-DT104 Salmonella strains did not possess any integrons. Two Salmonella isolates displayed tolerance to the organic solvent cyclohexane, indicating the possibility that they are overexpressing chromosomal regulatory genes marA or soxS or the associated multidrug efflux pump, acrAB. This research suggests that integrons contribute to antimicrobial resistance among specific swine Salmonella serotypes; however, they are not as widely disseminated among non-Typhimurium swine Salmonella serotypes as previously thought. PMID- 12528829 TI - Clonal distribution of an atypical MRP+, EF*, and suilysin+ phenotype of virulent Streptococcus suis serotype 2 strains in Brazil. AB - Streptococcus suis is considered one of the most important bacterial swine pathogens worldwide. The distribution of the 35 described serotypes in diseased animals may vary in different regions. Data regarding S. suis isolation from pigs in South America is not available. In the present study, 51 isolates of S. suis recovered in pure culture or as the predominant species from diseased animals in Brazil, were analyzed. These isolates were classified as serotypes 2 (58.8%), 3 (21.5%), 7 (13.7%), 1 (3.9%), and 14 (2%). Serotype 2 isolates were further studied for their production of virulence-related proteins muramidase-released protein (MRP), extracellular factor (EF), and suilysin. In addition, the genetic diversity was studied by randomly amplified polymorphic DNA. All but 1 of the serotype 2 isolates showed a clonal distribution of an atypical phenotype (MRP+, EF*, suilysin+), different from the known European (MRP+, EF+, suilysin+), and North American (MRPv, EF-, suilysin-), phenotypes. PMID- 12528830 TI - Evaluation of the effect of pH on in vitro growth of Malassezia pachydermatis. AB - The purpose of this study was to evaluate the effects of pH on the growth of canine Malassezia pachydermatis isolates in vitro. Yeast growth was monitored by measuring the optical density with a spectrophotometer. The growth of American Type Culture Collection and field strains of M. pachydermatis was optimal between the pH values of 4.0 and 8.0, and inhibited at the ranges of 1.0 to 3.0 and 9.0 to 10.0. An analysis of covariance showed no significant differences among the growth curves at pH levels 5.0 to 8.0. Although specific contrast tests showed that the growth slope at pH 4.0 was significantly different from that at pH 5.0 to 8.0, only small, random differences were found when the growth slope at pH 4.0 was compared to the individual slopes at pH 5.0, 6.0, 7.0, and 8.0. The findings of this study suggest that topical acidifying products could be beneficial therapeutic options for cutaneous yeast infections in dogs. PMID- 12528831 TI - Evaluation of the reliability and repeatability of automated milk urea nitrogen testing. AB - The purpose of this study was to evaluate the reliability (precision and accuracy), and repeatability of an infrared method of determining milk urea nitrogen. The reference method used for the reliability assessment was a wet chemistry, enzymatic determination of milk urea nitrogen. Reliability and repeatability, as measured by concordance correlation coefficients, were 0.97 and 0.99, respectively. PMID- 12528832 TI - The antagonistic effects of atipamezole and yohimbine on stress-related neurohormonal and metabolic responses induced by medetomidine in dogs. AB - This study aimed to compare the antagonistic effects of atipamezole (40,120, and 320 microg/kg, IM), yohimbine (110 microg/kg, IM), and saline on neurohormonal and metabolic responses induced by medetomidine (20 microg/kg, IM). Five beagle dogs were used in each of the 5 experimental groups in randomized order. Blood samples were taken for 6 h. Medetomidine significantly decreased norepinephrine, epinephrine, insulin, and nonesterified fatty acid levels, and increased plasma glucose levels. Both atipamezole and yohimbine antagonized these effects. The reversal effect of atipamezole was dose-dependency, except on epinephrine. Yohimbine caused prolonged increases in plasma norepinephrine and insulin levels compared to atipamezole, possibly because of its longer half-life elimination. Only yohimbine increased the cortisol levels. Neither glucagon nor lactate levels changed significantly. Based on these findings, when medetomidine-induced sedation is antagonized in dogs, we recommend using atipamezole IM, from 2- to 6 fold the dose of medetomidine, unless otherwise indicated. PMID- 12528834 TI - Temporal effects of freezing on plasma nitric oxide concentrations in ponies. AB - The purpose of this study was to compare concentrations of nitric oxide (NO) in fresh plasma versus frozen plasma, and determine the temporal effects of freezing on jugular venous plasma NO concentrations in clinically healthy ponies. Twenty eight helminth-naive ponies, aged from 4 to 6 mo, were raised and maintained under parasite-free conditions. Blood was collected from the jugular vein, centrifuged, and the plasma supernatant was analyzed fresh for NO concentrations using a chemiluminescent method. The remaining samples were aliquoted into 12 samples and stored at -70 degrees C until they were analyzed for NO concentration. Plasma NO concentration was measured at monthly intervals for 12 mo. There were significant differences in the plasma NO concentration across time compared with the baseline value at 1, 3, 4, 6, 7, and 11 mo. However, these values remained within the range for clinically healthy equids compared with concentrations of NO from horses with increased NO from diseased states. PMID- 12528833 TI - Effects of bisphenol-A on the growth of comb and testes of male chicken. AB - Bisphenol-A (BPA) has been reported to have some xenoestrogenic effects on the reproductive system of male animals. In this study, we examined the growth of combs and testes of the male chickens exposed to BPA. White leghorn male chicks were administered 200 mg BPA orally every week from 2 wk of age. The combs and testes were examined at 16 wk of age. The body growth showed no significant difference between BPA-administered and control birds. However, the weight of the combs and testes were lower in the BPA-treated birds. Histologically, testes of the control birds were well matured; the seminiferous tubuli were filled with sperm. In contrast, the testes of most of the BPA-treated birds showed an immature appearance with smaller seminiferous tubuli and limited spermatogenesis. These findings suggest that the xenoestrogenic property of BPA might disturb the growth of the comb and testes of male chickens by a possible endocrine disrupting mechanism. PMID- 12528836 TI - There's something about Medicaid. AB - In the thirty-seven years since its creation, Medicaid has grown in terms of whom it covers and what it costs. Current rates of Medicaid enrollment and cost growth are high relative to state budget capacity, but not by historical standards. The current Medicaid fiscal crisis is a result of weak state fiscal conditions and the gradual accretion of populations and services covered by Medicaid. States view Medicaid as an essential part of their current strategies to provide insurance to their low-income populations, cover the chronic care needs of people with disabilities and the elderly, and finance the health care safety net. Medicaid has accomplished much, and it can continue to do so if the underlying fiscal pressures and tensions built into it are addressed. PMID- 12528837 TI - Poor program's progress: the unanticipated politics of Medicaid policy. AB - Advocates of U.S. national health insurance tend to share an image that highlights universal standards of coverage, social insurance financing, and national administration--in short, the basic features of Medicare. Such an approach is said to be good (equitable and efficient) policy and equally good politics. Medicaid, by contrast, is often taken to exemplify poor policy and poorer politics: means-tested eligibility, general revenue financing, and federal/state administration, which encourage inequities and disparities of care. This stark juxtaposition fails, however, to address important counterintuitive elements in the political evolution of these programs. Medicare's benefits and beneficiaries have stayed disturbingly stable, but Medicaid's relatively broad benefits have held firm, and its categories of beneficiaries have expanded. Repeated alarms about "bankruptcy" have undermined confidence in Medicare's trust funding, while Medicaid's claims on the taxpayer's dollar have worn well. Medicare's national administration has avoided disparities, but at the price of sacrificing state and local flexibility that can ease such "reforms" as the introduction of managed care. That Medicaid has fared better than a "poor people's program" supposedly could has provocative implications for health reform debates. PMID- 12528838 TI - The bursting state fiscal bubble and state Medicaid budgets. AB - States make policy choices about Medicaid eligibility, benefits, and reimbursement rates in the context of the larger state fiscal environment. This paper reviews the factors that made this environment so favorable in the 1990s and examines the state budget outlook. State tax revenue is likely to grow more slowly in this decade than in the last, states will face continued substantial pressures to finance K-12 and higher education, and Medicaid spending has accelerated. States will have to curtail the growth of Medicaid, raise revenue, or constrain growth in education spending to maintain balanced budgets. PMID- 12528839 TI - The flexibility factor: finding the right balance. AB - Medicaid is financed with federal and state dollars and administered by states subject to certain federal rules. This design has strengths and weaknesses, and inherent tensions. Rising costs, state fiscal pressures, and new federal waiver policies have prompted some states to seek authority to use Medicaid dollars in ways that do not conform to current federal standards. These changes could have a sizable impact on the program and its beneficiaries. As federal and state roles and responsibilities are discussed, a range of options, including some that extend beyond Medicaid, will need to be considered to address the cost and resource issues that lie beneath much of the current debate. PMID- 12528840 TI - Medicaid and managed care: a lasting relationship? AB - The Medicaid program made a major commitment to managed care during the past decade. Following turbulent early years, the marriage matured and stabilized because managed care models responded well to a number of the states' goals and Medicaid purchasers were willing to make key trade-offs on behalf of their beneficiaries that conformed to the designs of managed care products. The relative tranquility in Medicaid managed care contrasts sharply with turmoil in both the commercial and Medicare sectors. But continuing changes in the managed care marketplace and financial distress in state budgets present new challenges to the strength and durability of this relationship. PMID- 12528841 TI - Where the action really is: Medicaid and the disabled. AB - Discussions of Medicaid tend to focus on low-income children and their mothers and the institutionalized elderly as the principal beneficiaries, but Medicaid spends more on the nonelderly disabled than on any other group. In the past two decades Medicaid has helped finance the deinstitutionalization of the mentally retarded and a growing proportion of the mentally ill, but implementation of the Olmstead decision has deflected advocates' attention from the more important issue of how managed care plans treat disabled Medicaid beneficiaries. PMID- 12528842 TI - Medicaid and mental health: be careful what you ask for. AB - Medicaid has had an enormous impact on the shape and impact of public mental health care. Medicaid mental health policy has expanded access, fostered consumerism, and created incentives for expansion of community-based providers. It also has dramatically changed the economic rules governing public mental health care, leading state governments to alter their behavior. The result has been a tilting of public mental health care toward Medicaid-covered people and services. PMID- 12528843 TI - The road to meaningful reform: a conversation with Oregon's John Kitzhaber. Interview by Jeff Goldsmith. PMID- 12528844 TI - An ethical template for pharmacy benefits. AB - We propose an ethical template for pharmacy benefits and a fair process for using it. The template delineates four levels of decisions about pharmacy coverage, connecting ethically acceptable types of rationales for limits with decisions made at each level. It provides a framework for organizing ethically relevant reasons for coverage (or the tiered copayments). The process for using the template assures accountability for the reasonableness of benefit decisions. It requires transparency and relevance of rationales for limit setting and revisability of decisions, including through fair procedures for appeals. The template and the process facilitate broader public learning about fair limit setting. PMID- 12528845 TI - Medicaid: lessons from a decade. AB - Drawing on a decade of analysis and examination of the Medicaid program, this paper assesses what we have learned about Medicaid--its role, its successes, and its setbacks. In the absence of universal coverage for health and long-term care, Medicaid is a critical component of our social safety net, taking on the highest risk, sickest, and often most expensive populations from private insurance and Medicare. Yet the substantial costs to federal and state governments incurred in filling this role, especially in lean economic times, remain its greatest challenge. PMID- 12528847 TI - Trends in U.S. health care spending, 2001. AB - U.S. health care spending grew 8.7 percent to $5,035 per capita in 2001. Total public funding continued to accelerate, increasing 9.4 percent and exceeding private funding growth by 1.2 percentage points. This acceleration was due in part to increased Medicaid spending in the midst of a recession and payment increases for Medicare providers. Prompted by sluggish economic growth and by faster-paced health spending, health spending's share of GDP spiked 0.8 percentage points in 2001 to 14.1 percent. PMID- 12528846 TI - The tenuous nature of the Medicaid entitlement. AB - Although Medicaid is regarded as a federal entitlement program, nowhere does the Medicaid statute explicitly recognize a federal right of action to enforce recipients' rights. Arguably, the Supreme Court, rather than Congress, first recognized the right of Medicaid recipients to protection of federal law. A controversial 2001 federal court decision, however, called into question the continuing existence of federally enforceable Medicaid rights. Although this decision has been reversed, it illuminates the tenuous nature of the Medicaid entitlement, as do recent Supreme Court decisions narrowing federal rights. Congress should amend the Medicaid statute to ensure the rights of Medicaid recipients. PMID- 12528848 TI - What drove private health insurance spending on mental health and substance abuse care, 1992-1999? AB - Trends in MH/SA treatment spending from 1992 to 1999 were examined using employer claims data from approximately 1.7 million covered lives in each year. The analysis finds that employer-based private insurance spending on MH/SA treatment did not keep pace with total employer-based private insurance spending or general price inflation. MH/SA spending dropped from 7.2 percent of total private insurance spending in 1992 to 5.1 percent in 1999. The decline was attributable to a dramatic decrease in inpatient MH/SA treatment--specifically, the probability of admissions and average length-of-stay. PMID- 12528849 TI - The role of public employers in a changing health care market. AB - Public employers provide health insurance coverage to nearly 16 percent of all U.S. workers. Their reactions to rapidly rising premiums can have an important effect on local markets for health insurance because of their size, their visibility, and their reflection of public policy. However, public employers are constrained in their responses by tight budgets set by elected officials and statutes regarding due process, public input, and public accountability. As insurance markets consolidate and premiums continue to increase, public employers face tough choices regarding employee benefits. PMID- 12528850 TI - How are health plans supporting physician practice? The physician perspective. AB - IPA-model HMOs are now the dominant organizational structures for delivering "managed care" in the United States. Are they taking advantage of opportunities to support physician practices in ways that arguably could improve care? In this paper we report the findings from a survey of generalist and specialist physicians in nineteen health plans. Not surprisingly, we found that generalists are much more likely than specialists are to be the target of health plans' efforts to support care delivery. However, our survey data indicate that these opportunities generally are not being fully exploited; also, efforts that plans do make to provide information to support care often are not seen as useful by physicians. PMID- 12528851 TI - The future supply of family physicians: implications for rural America. AB - Throughout the past century rural health care has been dependent upon general practitioners (GPs) and their successors, family physicians (FPs). Only FPs and GPs have practiced in rural areas in proportion to the population, then and now. As specialization occurred, numbers of GPs declined and physician shortages developed in rural areas. The creation of family practice residencies in the 1970s halted this decline, but rural shortages persist today. During the 1990s the number of allopathic and osteopathic FP residency graduates rose 54 percent. At the same time, the percentage of women enrolled in these residencies increased to 46 percent, and women have been less likely than men to select rural practice. We project that if current numbers of graduates continue, the nonmetropolitan FP/GP-to-population ratio will increase 17 percent by the year 2020. However, today, medical students' interest in primary care residencies (including family practice) is declining precipitously. If numbers of FP graduates return to 1993 levels, the density of FPs in rural America and in the nation as a whole will decline after 2010. PMID- 12528852 TI - Where's David? PMID- 12528853 TI - Medicaid managed care payment methods and capitation rates in 2001. AB - We present results from a survey of Medicaid managed care payment methods and rates in 2001 for AFDC/TANF and poverty-related Medicaid populations, updating a similar survey of 1998 rates. Rates were adjusted for differences in age-sex groupings, maternity payments, and service carve-outs. A twofold variation in Medicaid capitation rates remains, although there was a change in the composition of states at the top and bottom. The data also show that the growth in Medicaid capitation rates between 1998 and 2001 averaged 18 percent, considerably more than the increase in Medicare+Choice rates. PMID- 12528854 TI - Transitions in insurance coverage from before pregnancy through delivery in nine states, 1996-1999. AB - Efforts to extend coverage to pregnant women, along with an expanding economy, did not prevent increases in the uninsured in the latter 1990s. Welfare reform may have led to declining Medicaid enrollments and caseloads. Data representative of live births in nine states show that in some states more than one-third of all pregnant women and almost two-thirds of low-income pregnant women lacked insurance before their pregnancy in 1996 and 1999. More than one-third of all pregnant women made some change in coverage by the time they delivered their baby. Among low-income women, the largest change was from uninsured status before pregnancy to Medicaid at delivery. PMID- 12528855 TI - Participation of plans and providers in Medicaid and SCHIP managed care. State Children's Health Insurance Program. AB - For Medicaid and SCHIP managed care programs to succeed, they must attract enough and the right kinds of plans and providers to meet access and care goals. In 2001 we analyzed practices and perceptions that bear on these goals by surveying managed care plans participating in Medicaid or SCHIP, or both, in eleven states. Participating plans appear supportive of both programs and are largely able to secure providers to participate, too. To date, SCHIP has not attracted many plans not already participating in Medicaid. While perceptions were positive in 2001, maintaining current plan and provider relationships in an environment that has become much more budget constrained will be challenging. PMID- 12528856 TI - Why is there state variation in employer-sponsored insurance? AB - Using the National Survey of America's Families in 1997 and 1999, we investigate the sources of variation in employer-sponsored health insurance across states. We find that demographics and family characteristics (such as race/ethnicity and citizenship status), individual employment characteristics (such as firm size and labor-force attachment), and local labor market characteristics (such as unionization) consistently explain the relative position of all of the states with either high or low rates of employer coverage. Income plays a smaller role in explaining the state variation but is still an important determinant, especially among states whose average income is far from the national average. PMID- 12528857 TI - Radical simplification: Disaster Relief Medicaid in New York City. AB - In the four months following the 11 September 2001 attacks on the World Trade Center, nearly 350,000 people signed up for Disaster Relief Medicaid. The process was quick and simple; applicants completed a one-page form and got a decision on the spot or the next day. While the program's success stemmed in part from the unique circumstances facing New Yorkers in the fall of 2001, Disaster Relief Medicaid was an experiment in radical simplification that demonstrated a new way of thinking about how to design a simple, effective public health insurance program stripped of the vestiges of welfare. PMID- 12528858 TI - Models and momentum for insuring low-income, undocumented immigrant children in California. AB - Undocumented immigrant children are an underserved, vulnerable population that has not benefited from the recent expansion of publicly funded children's health insurance programs. The California Endowment funded a two-year demonstration project to provide subsidized health insurance coverage to more than 7,500 children through five nonprofit organizations. Sustaining and promoting coverage for this population will require continuing subsidies through a mix of private and public funding. Locally based, comprehensive initiatives are in place or emerging in a growing list of California counties. PMID- 12528859 TI - Viagra and contraceptives. PMID- 12528860 TI - Air pollution and medical care. PMID- 12528861 TI - Rational public policy. PMID- 12528862 TI - Health insurance system promotes disparities. PMID- 12528863 TI - Consumer-driven health plans. PMID- 12528864 TI - Male and female nurse dissatisfaction. PMID- 12528865 TI - Nurse staffing. PMID- 12528866 TI - Benefits of osteopathic schools. PMID- 12528867 TI - Market demand matters. PMID- 12528868 TI - Moonlighting residents. PMID- 12528869 TI - Liposuction procedures: how, not where. PMID- 12528870 TI - Burden-of-disease estimates. PMID- 12528871 TI - Henry Hallett Dale--chemical transmission of nerve impulses. PMID- 12528872 TI - Multiple myeloma: how far have we come? PMID- 12528873 TI - Assessment of T-wave morphology. PMID- 12528874 TI - Review of 1027 patients with newly diagnosed multiple myeloma. AB - OBJECTIVE: To determine the clinical and laboratory features of newly diagnosed multiple myeloma. PATIENTS AND METHODS: Records of all patients in whom multiple myeloma was initially diagnosed at the Mayo Clinic in Rochester, Minn, from January 1, 1985, to December 31, 1998, were reviewed. RESULTS: Of the 1027 study patients, 2% were younger than 40 years, and 38% were 70 years or older. The median age was 66 years. Anemia was present initially in 73% of patients, hypercalcemia (calcium level > or = 11 mg/dL) in 13%, and a serum creatinine level of 2 mg/dL or more in 19%. The beta2-microglobulin level was increased in 75%. Serum protein electrophoresis revealed a localized band in 82% of patients, and immunoelectrophoresis or immunofixation showed a monoclonal protein in 93%. A monoclonal light chain was found in the urine in 78%. Nonsecretory myeloma was recognized in 3% of patients, whereas light-chain myeloma was present in 20%. Conventional radiographs showed an abnormality in 79%. The plasma cell labeling index was 1% or more in 34% of patients. Multivariate analysis revealed that age, plasma cell labeling index, low platelet count, serum albumin value, and the log of the creatinine value were the most important prognostic factors. CONCLUSION: The median duration of survival was 33 months and did not improve from 1985 through 1998. PMID- 12528876 TI - Catecholamine-induced T-wave lability in congenital long QT syndrome: a novel phenomenon associated with syncope and cardiac arrest. AB - OBJECTIVE: To determine the effects of phenylephrine and dobutamine on repolarization lability in patients with genotyped long QT syndrome (LQTS). PATIENTS AND METHODS: Between December 1998 and August 2000, 23 patients with genotyped LQTS (13 LQT1, 7 LQT2, and 3 LQT3) and 16 controls underwent electrocardiographic stress testing at the Mayo Clinic in Rochester, Minn. Aperiodic repolarization lability was quantified from digitized electrocardiograms recorded during catecholamine stress testing with phenylephrine and dobutamine. T-wave lability was quantified as a root-mean square of the differences between corresponding signal values of subsequent beats. The magnitude of aperiodic T-wave lability was quantified by using a newly derived T-wave lability index (TWLI). RESULTS: The TWLI was significantly greater in patients with LQTS than in controls (0.0945 +/- 0.0517 vs 0.0445 +/- 0.0123; P < .003). Marked T-wave lability (TWLI > or = 0.095) was detected in all 3 LQTS genotypes (10/23) but in no controls (P < .003). There was no correlation between the TWLI and the baseline corrected QT interval. All high-risk patients having either a history of out-of-hospital cardiac arrest or syncope had a TWLI of 0.095 or greater. CONCLUSIONS: Beat-to-beat nonalternating T-wave lability occurs in LQT1, LQT2, and LQT3 patients during catecholamine provocation and is associated with a history of prior cardiac events. The quantification of this novel phenomenon may assist in identifying LQTS patients with increased risk of sudden cardiac death. PMID- 12528875 TI - Response rate, durability of response, and survival after thalidomide therapy for relapsed multiple myeloma. AB - OBJECTIVE: To assess response rate, duration of response, progression-free survival, and toxicity of thalidomide in patients with relapsed multiple myeloma. PATIENTS AND METHODS: Thirty-two patients with relapsed multiple myeloma were entered into the study between April 29, 1999, and June 20, 2000. They were given oral thalidomide at a dosage of 200 mg/d for 2 weeks, which was then increased as tolerated to a maximum of 800 mg/d. The primary end point of the study was response rate. RESULTS: The median age of the patients was 67 years (range, 36-78 years). Prior chemotherapy had failed in all patients, and stem cell transplantation had failed in 5 patients (16%). There were 10 confirmed responses, yielding a response rate of 31%. In addition, there was 1 unconfirmed partial response and 7 minimal responses with no complete responses. The median duration of response was 11.9 months (range, 3.7-203 months). Overall, 20 patients have died, and 26 patients have experienced disease progression. The median follow-up of surviving patients was 28.5 months (range, 193-34.0 months), with a median progression-free survival of 8.6 months (95% confidence interval [CI], 4.7-16 months). The median progression-free survival among the responding patients was 15.7 months (95% CI, 8.6-25.6 months). The median overall survival for the entire group was 22 months (95% CI, 10.6-35.9 months). The most common treatment-related nonhematologic toxic effects (grade >3) were neuropathy (16%), sedation (13%), febrile neutropenia (6%), and constipation (6%). CONCLUSIONS: Thalidomide is useful in the treatment of patients with relapsed multiple myeloma and produced durable response in approximately one third of patients, with median response duration of nearly 1 year. PMID- 12528877 TI - Multiple blood donations associated with iron deficiency in patients with restless legs syndrome. AB - OBJECTIVE: To describe a series of patients with restless legs syndrome (RLS) and iron deficiency with and without anemia related to repeated blood donations. PATIENTS AND METHODS: Study patients were identified by asking consecutive patients with RLS seen at the Mayo Clinic in Rochester, Minn, from February 1 to December 31, 2001, whether they donated blood. All patients who fulfilled the International Restless Legs Syndrome Study Group criteria for RLS, had donated blood a minimum of 3 times a year the preceding 3 years, and had iron deficiency (serum ferritin concentration <20 microg/L) were included in the study. RESULTS: Eight patients met the study criteria. The mean +/- SD serum ferritin concentration was 8.1 +/- 3.5 microg/L, and 4 patients had anemia. In 6 of the 8 patients, RLS began at about the same time of or after blood donation. Patients had donated blood for 4.2 +/- 13 times a year (range, 3-6 times a year) for 15.2 +/- 83 years (range, 5-25 years). Hemoglobin concentrations were 12.8 +/- 1.8 g/dL (range, 10.6-15.5 g/dL). In 2 patients, RLS essentially resolved with correction of iron stores alone, and medications for RLS were successfully discontinued in 2 other patients. CONCLUSIONS: Repeated blood donation is associated with induction or perpetuation of RLS due to iron deficiency with or without coexisting anemia. Potential blood donors should be questioned about RLS, and donation should not be allowed until the serum ferritin concentration has been measured and iron stores replenished if necessary. PMID- 12528878 TI - Primer on medical genomics part V: bioinformatics. AB - Bioinformatics is the discipline that develops and applies informatics to the field of molecular biology. Although a comprehensive review of the entire field of bioinformatics is beyond the scope of this article, I review the basic tenets of the field and provide a topical sampling of the popular technologies available to clinicians and researchers. These technologies include tools and methods for sequence analysis (nucleotide and protein sequences), rendering of secondary and tertiary structures for these molecules, and protein fold prediction that can lead to rational drug design. I then discuss signaling pathways, new standards for data representation of genes and proteins, and finally the promise of merging these molecular data with the clinical world (the new science of phenomics). PMID- 12528879 TI - The patient-computer interview: a neglected tool that can aid the clinician. AB - In this article, I (1) review the process of interviewing patients by computer, (2) summarize computer-interviewing work done in 1968, (3) address the weaknesses of collecting information with the traditional history-taking methods or paper questionnaires, (4) discuss commercial software designed for computer interviewing, and (5) focus on the strengths and weaknesses of interviewing patients with a computer. The strengths of this process compared with traditional interviewing are that computer interviewing allows the physician to gather more data; gives the patient more time to complete an interview; uncovers more sensitive information; provides more adaptability to non-English-speaking patients, patients with hearing impairment, or patients who are illiterate; and provides structured information for research. The weaknesses of computer interviewing are that it generates false-positive responses, is not accepted by a minority of patients, is unable to detect nonverbal behavior, and requires changes in work flow. With the advent of an electronic medical record and the financial rewards for comprehensive history recording, the gathering of history and documentation from patients is increasingly important and favors adaptation to computer interviewing. PMID- 12528880 TI - Evolving role of the neurologist in the diagnosis and treatment of chronic noncancer pain. AB - The neurologist has become increasingly involved in the multidisciplinary treatment of patients with chronic noncancer pain (CNP). Chronic noncancer pain affects a diverse patient population with multiple underlying diagnoses and associated therapies. Following the model of the American Board of Anesthesiology and the American Society of Anesthesiologists for practice guidelines and subspecialty requirements, neurologic pain management is now recognized as a subspecialty of neurology by the American Academy of Neurology and the American Board of Psychiatry and Neurology. Current basic and clinical research into the neuropathology, neurophysiology, neurochemistry, and neuropharmacology of chronic pain continues to expand diagnostic and therapeutic options. Informed regulatory agencies and professional organizations such as the American Academy of Neurology recognize the undertreatment of patients with CNP and provide clear recommendations to help neurologists in the ethical and effective treatment of patients with pain. Improved education of neurologists, other health care professionals, patients, and the media about evolving standards of pain care and therapy will produce a more supportive environment for the compassionate and ethical treatment of patients with CNP. PMID- 12528881 TI - Searching the medical literature using PubMed: a tutorial. AB - Staying current with advances in medicine is becoming a major challenge for clinicians. Access to updated repositories of medical information and the expertise to locate relevant information within them are becoming necessary clinical skills. PubMed (http://www.pubmedgov) provides free access to the largest biomedical resource available and is updated daily. Clinicians can use this resource to find answers to focused clinical questions quickly and efficiently. The purpose of this article is to assist clinicians in the development of the basic skills required to use PubMed to make informed clinical decisions. PMID- 12528882 TI - Economic and psychological implications of the obesity epidemic. PMID- 12528883 TI - 27-Year-old woman with numbness and weakness of the extremities. PMID- 12528884 TI - Systemic thrombolytic therapy after recent abdominal aortic aneurysm repair: an absolute contraindication? AB - Systemic thrombolysis in the early postoperative period can cause fatal hemorrhage. Systemic thrombolysis is often considered contraindicated after major vascular procedures; thus, experience with this scenario is limited. A 67-year old man experienced massive pulmonary embolization after his abdominal aortic aneurysm was repaired with a bifurcated, woven Dacron graft. Because systemic thrombolysis was the only option for our patient's survival, he underwent this procedure with recombinant tis-sue-type plasminogen activator just 2 weeks after the Dacron graft repair of his abdominal aortic aneurysm. After clinical stabilization, abdominal and pelvic computed tomography showed no periprosthetic graft hemorrhage. The successful systemic thrombolysis suggests that this therapy may prove useful in extreme situations. PMID- 12528885 TI - Paradoxical embolism in the left main coronary artery: diagnosis by transesophageal echocardiography. AB - We describe a patient with a paradoxical coronary embolism diagnosed by transesophageal echocardiography. The patient developed a stroke followed by a myocardial infarction. Coronary angiography showed an obstruction of the left main coronary artery. Transesophageal echocardiography showed the mechanism of the neurologic and cardiac events to be a paradoxical embolism. Emergency surgical retrieval of the thrombus lodged in the left main coronary ostium and of a separate thrombus traversing a patent foramen ovale was performed. To our knowledge, direct visualization of the paradoxical coronary embolism by echocardiography has not been reported previously. We discuss mechanisms responsible for paradoxical coronary embolism and review the literature pertaining to this condition. PMID- 12528886 TI - An overview of surgery for chronic seizures. AB - Intracranial surgical intervention offers the potential to reduce or eliminate seizures in a substantial number of patients with medically intractable epilepsy. Although some patients with epilepsy have a pathologic condition that can be imaged radiographically, this is not a requirement for the patient to be considered a surgical candidate. Evaluation in a comprehensive epilepsy center includes defining the characteristics of the seizures; integrating medical criteria, psychosocial factors, imaging findings, and surgical risks; and weighing these factors against the risks of continued, uncontrolled epilepsy. Several neurosurgical procedures are available to help these patients, including frontal and temporal lobectomy, lesionectomy, subpial transection, and callosotomy. PMID- 12528887 TI - Use of bisphosphonates in patients with myeloma and renal failure. PMID- 12528888 TI - Quality of adverse event assessment in randomized controlled trials and observational studies. PMID- 12528889 TI - The role of nitric oxide in cancer. AB - Nitric oxide (NO) is a pleiotropic regulator, critical to numerous biological processes, including vasodilatation, neurotransmission and macrophage-mediated immunity. The family of nitric oxide synthases (NOS) comprises inducible NOS (iNOS), endothelia (eNOS), and neuronal NOS (nNOS). Interestingly, various studies have shown that all three isoforms can be involved in promoting or inhibiting the etiology of cancer. NOS activity has been detected in tumour cells of various histogenetic origins and has been associated with tumour grade, proliferation rate and expression of important signaling components associated with cancer development such as the oestrogen receptor. It appears that high levels of NOS expression (for example, generated by activated macrophages) may be cytostatic or cytotoxic for tumor cells, whereas low level activity can have the opposite effect and promote tumour growth. Paradoxically therefore, NO (and related reactive nitrogen species) may have both genotoxic and angiogenic properties. Increased NO-generation in a cell may select mutant p53 cells and contribute to tumour angiogenesis by upregulating VEGF. In addition, NO may modulate tumour DNA repair mechanisms by upregulating p53, poly(ADP-ribose) polymerase (PARP) and the DNA-dependent protein kinase (DNA-PK). An understanding at the molecular level of the role of NO in cancer will have profound therapeutic implications for the diagnosis and treatment of disease. PMID- 12528890 TI - Stem cell pluripotency and transcription factor Oct4. AB - Mammalian cell totipotency is a subject that has fascinated scientists for generations. A long lasting question whether some of the somatic cells retains totipotency was answered by the cloning of Dolly at the end of the 20th century. The dawn of the 21st has brought forward great expectations in harnessing the power of totipotentcy in medicine. Through stem cell biology, it is possible to generate any parts of the human body by stem cell engineering. Considerable resources will be devoted to harness the untapped potentials of stem cells in the foreseeable future which may transform medicine as we know today. At the molecular level, totipotency has been linked to a singular transcription factor and its expression appears to define whether a cell should be totipotent. Named Oct4, it can activate or repress the expression of various genes. Curiously, very little is known about Oct4 beyond its ability to regulate gene expression. The mechanism by which Oct4 specifies totipotency remains entirely unresolved. In this review, we summarize the structure and function of Oct4 and address issues related to Oct4 function in maintaining totipotency or pluripotency of embryonic stem cells. PMID- 12528891 TI - cAMP elevators inhibit LPS-induced IL-12 p40 expression by interfering with phosphorylation of p38 MAPK in murine peritoneal macrophages. AB - cAMP mediated signaling may play a suppressive role in immune response. We previously found that the cAMP-elevators (CTx and 8-Br-cAMP) inhibited IL-12, IL la, IL-6 gene expression, but increased the transcriptional levels of IL-10 and IL-1Ra in LPS-treated murine peritoneal macrophages. The present study examined a possible molecular mechanism involved in cAMP elevators-induced inhibition of IL 12 p40 expression in response to LPS. Our data demonstrated that cAMP elevators downregulated IL-12 p40 mRNA expression and IL-12 p70 production in murine peritoneal macrophages. Subsequent studies revealed that cAMP-elevators blocked phosphorylation of p38 MAPK, but did not affect the activity of NF-kappaB binding to IL-12 promoter (-136/-112). This is the first report that cAMP elevators inhibit LPS-induced IL-12 production by a mechanism that is associated, at least in part, with p38-dependent inhibition by cAMP signaling pathways. PMID- 12528892 TI - The promoter analysis of the human C17orf25 gene, a novel chromosome 17p13.3 gene. AB - The human C17orf25 gene (Accession No. AF177342) is one of thirteen genes cloned from a region displaying a high score of loss of heterozygosity within chromosome 17p13.3 in human hepatocellular carcinoma in China. To unveil the underlying mechanisms for the transcription regulation of this gene and understand its implication to the hepatocellular carcinogenesis, we looked into the relevant aspects by both bioinformatic and experimental executions. We found: 1, The abundant expression of the C17orf25 gene was evident in all the cell lines and tissue samples tested, showing little hepatoma-selectivity; 2, Its transcription starts at a single site, locating at -60 from the translation initiation codon; 3, A 58 bp fragment containing the transcription start, extending from -112 to 55, represents the minimal promoter; 4, The consensus sequence within this fragment recognized by SP1 contributes predominantly to the activity of the minimal promoter; 5, The bioinformatic analysis suggests that the C17orf25 gene may encode a protein in the family of the glyoxalase. Our data has provided some deep insight into both function and regulation of the C17orf25 gene in the context of the normal liver and hepatocellular carcinoma. PMID- 12528893 TI - Cloning and characterization of a mouse liver-specific gene mfrep-1, up-regulated in liver regeneration. AB - Human fibrinogen-related protein-1/liver fibrinogen-related protein-1 (HFREP 1/LFIRE-1), a liver-specific protein, is a member of fibrinogen superfamily that exerts various biological activities. However, the function of HFREP-1/LFIRE-1 in liver remains unknown. Here we isolated its mouse ortholog gene-mouse fibrinogen related protein-1 (mfrep-1), which encoded 314 amino acids, exhibiting 80.4% similarity to HFREP-1/LFIRE-1. Northern blot analysis revealed that 1.2-kb mfrep 1 mRNA was detected selectively in mouse liver. To explore the function of MFREP 1, we examined the levels of mfrep-1 mRNA during regeneration after 70% partial hepatectomy (PHx) in mice. mfrep-1 mRNA increased in the regenerating liver and reached the first shoulder peak at 2-4 h after PHx. Cycloheximide pretreatment could suppress the induction of mfrep-1, indicating the up-regulation of this gene need de novo protein synthesis. Its mRNA continued to elevate at 6 h thereafter and reached the second peak at 24 h. The enhanced expression of mfrep 1 maintained high until 72 h and then declined slowly to the basal level. Immunohistochemistry assessment confirmed the up-regulated expression of MFREP-1 protein in parenchymal cells during liver regeneration. These data suggested that MFREP-1 might play an important role in liver regeneration and be involved in the regulation of cell growth. PMID- 12528894 TI - The associated regulators and signal pathway in rIL-16/CD4 mediated growth regulation in Jurkat cells. AB - IL-16 is a ligand and chemotactic factor for CD4+ T cells. IL-16 inhibits the CD3 mediated lymphocyte activation and proliferation. The effects of IL-16 on the target cells are dependent on the cell type, the presence of co-activators etc. To understand the regulation function and mechanism of IL-16 on target cells, we used a 130 a.a. recombinant IL-16 to study its effects on the growth of Jurkat T leukemia cells in vitro. We found that the rIL-16 stimulated the proliferation of Jurkat cells at low dose (10(-9)M), but inhibited the growth of the cells at higher concentration (10(-5)M). Results showed that 10(-5) M of rIL-16 treatment induced an enhanced apoptosis in Jurkat cells. The treatment blocked the expression of FasL, but up-regulated the c-myc and Bid expression in the cells. Pre-treatment of PKC inhibitor or MEK1 inhibitor markedly increased or decreased the rIL-16 induced growth-inhibiting effects on Jurkat cells, respectively. The results suggested that the rIL-16 might be a regulator for the growth or apoptosis of Jurkat cells at a dose-dependent manner. The growth-inhibiting effects of rIL-16 might be Fas/FasL independent, but, associated with the activation of PKC, up-regulated expression of c-Myc and Bid, and the participation of the ERK signal pathway in Jurkat cells. PMID- 12528895 TI - Regulation of cellular adhesion molecule expression in murine oocytes, peri implantation and post-implantation embryos. AB - Expression of the adhesion molecules, ICAM-1, VCAM-1, NCAM, CD44, CD49d (VLA-4, alpha chain), and CD11a (LFA-1, alpha chain) on mouse oocytes, and pre- and peri implantation stage embryos was examined by quantitative indirect immunofluorescence microscopy. ICAM-1 was most strongly expressed at the oocyte stage, gradually declining almost to undetectable levels by the expanded blastocyst stage. NCAM, also expressed maximally on the oocyte, declined to undetectable levels beyond the morula stage. On the other hand, CD44 declined from highest expression at the oocyte stage to show a second maximum at the compacted 8-cell/morula. This molecule exhibited high expression around contact areas between trophectoderm and zona pellucida during blastocyst hatching. CD49d was highly expressed in the oocyte, remained significantly expressed throughout and after blastocyst hatching was expressed on the polar trophectoderm. Like CD44, CD49d declined to undetectable levels at the blastocyst outgrowth stage. Expression of both VCAM-1 and CD11a was undetectable throughout. The diametrical temporal expression pattern of ICAM-1 and NCAM compared to CD44 and CD49d suggest that dynamic changes in expression of adhesion molecules may be important for interaction of the embryo with the maternal cellular environment as well as for continuing development and survival of the early embryo. PMID- 12528896 TI - Developmental changes in functional expression and beta-adrenergic regulation of I(f) in the heart of mouse embryo. AB - The hyperpolarization-activated current (I(f)) plays an important role in determining the spontaneous rate of cardiac pacemaker cells. The automatic rhythmicity also exists in working cells of embryonic heart, therefore we studied developmental changes in functional expression and beta-adrenergic regulation of I(f) in embryonic mouse heart. The expression of I(f) is high in early developmental stage (EDS) (10.5 d after coitus) ventricular myocytes, low in intermediate developmental stage (IDS) (13.5 d) atrial or ventricular myocytes and even lower in late developmental stage (LDS) (16.5 d) atrial or ventricular myocytes, indicating that these cells of the EDS embryonic heart have some properties of pacemaker cells. Beta-adrenergic agonist isoproterenol (ISO) stimulates I(f) in LDS but not in EDS cardiomyocytes, indicating that the beta adrenergic regulation of I(f) is not mature in EDS embryonic heart. But forskolin (a direct activator of adenylate cyclase) and 8-Br-cAMP (a membrane-permeable analogue of cAMP) increase the amplitude of I(f) in EDS cells, indicating that adenylate cyclase and cAMP function fairly well at early stage of development. Furthermore, the results demonstrate that I(f) is modulated by phosphorylation via cAMP dependent PKA both in EDS and LDS cells. PMID- 12528897 TI - Increased exchange rate of histone H1 on chromatin by exogenous myogenin expression. AB - To explore the molecular mechanism of chromatin remodeling involved in the regulation of transcriptional activation of specific genes by a myogenic regulatory factor Myogenin, we used NIH3T3 fibroblasts with a stably integrated H1.1-GFP fusion protein to monitor histone H1 movement directly by fluorescence recovery after photobleaching (FRAP) in living cells. The observation from FRAP experiments with myogenin transfected fibroblasts showed that the exchange rate of histone H1 in chromatin was obviously increased, indicating that forced expression of exogenous Myogenin can induce chromatin remodeling. The hyperacetylation of histones H3 and H4 from myogenin transfected fibroblasts was detected by triton-acid-urea (TAU)/SDS (2-D) electrophoresis and Western blot with specific antibodies against acetylated N-termini of histones H3 and H4. RT PCR analysis indicated that the nAChR alpha-subunit gene was expressed in the transfected fibroblasts. These results suggest that the expression of exogenous Myogenin can induce chromatin remodeling and activate the transcription of Myogenin-targeted gene in non-muscle cells. PMID- 12528898 TI - Preparation and analysis of spermatocyte meiotic pachytene bivalents of pigs for gene mapping. AB - Well-spread meiotic pachytene bivalents were obtained by using the prolonged hypotonic treatment combined with high chloroform Carnory's fixative solution from cells of the testes of domestic pigs. Comparison in the division index and length of pachytene bivalents with metaphase chromosomes showed that those of the former are 5 times higher and 3.42(1.87-5.98) times longer than those of the latter. Comparative studies on chromomere maps of bivalents and mitotic chromosomal G-bands were conducted by using the chromosome 12 as a example. Sex vesicle and various shapes of synaptic sex chromosomes have been observed. Two color PRimed IN Situ (PRINS) labeling has been conducted successfully on pachytene bivalents of pigs. PMID- 12528899 TI - Identification and characterization of a new member of serpin family- HongrES1 in rat epididymis. AB - A full length cDNA named HongrES1 was isolated and cloned by screening rat epididymis cDNA library using a mouse EST as a probe and 5'RACE followed. It contained 1590bp nucleotides and its predicted protein had 415 amino acid residues including a serpin (serine protease inhibitor) conserved domain. Tissue distribution pattern showed it was specifically expressed in adult rat epididymis; moreover, in situ hybridyzation indicated this gene was expressed in a limited region of the cauda epididymis near vas deference. Such kind of expression pattern sugested that HongrES1 had potential function in male reproduction. PMID- 12528901 TI - A cognitive complexity metric applied to cognitive development. AB - Two experiments tested predictions from a theory in which processing load depends on relational complexity (RC), the number of variables related in a single decision. Tasks from six domains (transitivity, hierarchical classification, class inclusion, cardinality, relative-clause sentence comprehension, and hypothesis testing) were administered to children aged 3-8 years. Complexity analyses indicated that the domains entailed ternary relations (three variables). Simpler binary-relation (two variables) items were included for each domain. Thus RC was manipulated with other factors tightly controlled. Results indicated that (i) ternary-relation items were more difficult than comparable binary-relation items, (ii) the RC manipulation was sensitive to age-related changes, (iii) ternary relations were processed at a median age of 5 years, (iv) cross-task correlations were positive, with all tasks loading on a single factor (RC), (v) RC factor scores accounted for 80% (88%) of age-related variance in fluid intelligence (compositionality of sets), (vi) binary- and ternary-relation items formed separate complexity classes, and (vii) the RC approach to defining cognitive complexity is applicable to different content domains. PMID- 12528900 TI - The interaction between the human beta-globin locus control region and nuclear matrix. AB - Our previous study showed that hydroxyurea (Hu) could induce HEL cells to express human beta-globin gene. However the molecular mechanisms by which the expression of beta-globin gene is activated and regulated are poorly understood. Here we show that the binding patterns between the core DNA sequences (HS2 core sequence 10681 approximately -10971 bp, HS3 core sequence -14991 approximately -14716 bp and HS4 core sequence -18586 approximately -18306 bp) of DNase I hypersensitive sites in the human beta-globin LCR and nuclear matrix proteins isolated from Hu induced and uninduced HEL cells are quite different. Results demonstrated that nuclear matrix proteins might play important roles in regulating the expression of human beta-like globin genes through their interaction with HSs (HS2, HS3 and HS4 core sequences) in the LCR. Moreover, the results obtained from the in vitro DNA-matrix binding assay showed that the core DNA sequences of DNase I hypersensitive sites (HS2, HS3 and HS4) were unable to bind to the nuclear matrix isolated from uninduced HEL cells; in addition, HS2 core DNA sequence was capable of binding to the nuclear matrix prepared from Hu-induced HEL cells, while both HS3 and HS4 core DNA sequences could not do so. Results indicated that the HS2 core DNA sequence may be a functional MAR (matrix attachment region). We suggest that the HS2 core DNA sequence binding to the nuclear matrix in Hu-induced HEL cells may open the structure of chromatin to make the LCR accessible to the promoter of beta-globin gene and to promote its transcription. PMID- 12528902 TI - Representation and competition in the perception of spoken words. AB - We present data from four experiments using cross-modal priming to examine the effects of competitor environment on lexical activation during the time course of the perception of a spoken word. The research is conducted from the perspective of a distributed model of speech perception and lexical representation, which focuses on activation at the level of lexical content. In this model, the strength of competition between simultaneously active lexical items depends on the degree of coherence between their distributed semantic and phonological representations. Consistent with this model, interference effects are more complete when the purely semantic aspects of these coactive representations are probed (using semantic priming) than when phonological aspects are probed as well (using repetition priming). PMID- 12528904 TI - Anatomy and biomechanics of the anterior cruciate ligament. AB - This article reviews the literature on embryology, anatomy, function, and biomechanics to define the properties of the native ACL. The discussion focuses on the ligamentous architecture and gross anatomy of the ACL including its femoral and tibial insertions as well as its midsubstance passing through the intercondylar fossa. Biomechanical modeling, mechanical and structural properties, and function of the ACL as a primary and secondary stabilizer under non-weightbearing and weightbearing conditions are described. PMID- 12528903 TI - Developments in young infants' reasoning about occluded objects. AB - Eight experiments were conducted to examine 3- and 3.5-month-old infants' responses to occlusion events. The results revealed two developments, one in infants' knowledge of when objects should and should not be occluded and the other in infants' ability to posit additional objects to make sense of events that would otherwise violate their occlusion knowledge. The first development is that, beginning at about 3 months of age, infants expect an object to become temporarily visible when passing behind an occluder with an opening extending from its lower edge. The second development is that, beginning at about 3.5 months of age, infants generate a two-object explanation when shown a violation in which an object fails to become visible when passing behind an occluder with an opening in its lower edge. Unless given information contradicting such an explanation, infants infer that two identical objects are involved in the event, one traveling to the left and one to the right of the opening. These and related findings provide the basis for a model of young infants' responses to occlusion events; alternative models are also discussed. PMID- 12528905 TI - Fate of the anterior cruciate ligament-injured knee. AB - Most patients with anterior cruciate ligament (ACL) injuries do well with activities of daily living even after follow-up in the range of 5 to 15 years. Most can participate in some sports activity if they are inclined to do so, but most will have some limitations in vigorous sports, and only a few will be entirely asymptomatic. The challenge to the clinician is to understand and predict how ACL deficiency in a given patient will affect that patients's life and activities. In counseling patients about treatment after an ACL injury, the clinician can use knee ligament arthrometry measurements and pre-injury sports activity to estimate the risk of injury over the next 5 to 10 years. Meniscus, chondral, and sub-chondral injuries are not uncommon, but rarely require surgical intervention in the early phase of ACL deficiency. The prevalence of clinically significant meniscal damage increases with time, and is associated with increasing disability, surgery, and arthrosis in high-risk patients. Ligament reconstruction has not been shown to prevent arthrosis, but in prospective studies it appears to reduce the risk of subsequent meniscal injury, improve passive anteroposterior knee motion limits, and facilitate return to high-level sporting activities. PMID- 12528906 TI - The female ACL: why is it more prone to injury? AB - Multiple factors are responsible for ACL tears. The key factor in the gender discrepancy appears to be dynamic, not static, and proximal, not distal. The factors involved in evaluating the female ACL are multiple. However, it is the dynamic movement patterns ot hip and knee position with increased flexion and a coordinated proximal muscle firing pattern to keep the body in a safe landing position that are the most critical factors. An ACL injury at an early age is a life-changing event. We can very successfully reconstruct and rehabilitate an ACL, but we cannot stop there. We must now go into the prevention arena. In the United States there is tremendous variation in the exposure and acquisition of skills of physical activities in our youth. Today, children are often playing inside, using computers and watching television-missing out on the opportunity to learn safe movement patterns. Therefore, physical movement classes should occur very early in life, teaching children to land safely and in control, similar to the cry of "get down, stay down" routinely heard during youth soccer. Similarly, specific strength training programs can address landing as well as foot movements during cutting in basketball. Coaches should issue stern warnings when athletes demonstrate a high-risk movement patterns such as one-leg landings, out-of control baseline landings, or straight-leg landings. The warnings may serve to keep the athlete from "touching the hot stove again" for fear of getting burned. No athlete feels she will be the one to get injured. Therefore, prospective analysis is likely to be received more warmly by the athletes if the program is presented with an emphasis on performance improvement rather than injury prevention. With increased participation in these programs, multiple-center analysis will have the power necessary to determine which factors significantly predispose athletes to ACL injury. The future for injury prevention is bright. We must rise to the challenge. PMID- 12528907 TI - Future direction of the treatment of ACL ruptures. AB - The future of treatment of the ACL rupture is changing as our understanding of the biology surrounding the ACL continues to increase. It is our expectation that clinically applicable treatments, including the repair of the ACL and the development of a biologically engineered ACL, will occur in the next decade. PMID- 12528908 TI - Imaging the anterior cruciate ligament. AB - MR imaging has surpassed all other imaging modalities to become the "gold standard" for imaging evaluation of ACL injury. The accuracy and sensitivity of MR imaging for evaluation of ACL tears is excellent when correlated with clinical tests and arthroscopic findings, and is improved in equivocal cases with assessment of secondary signs for ACL tear. The MR imaging assessment of partial and chronic ACL tears is less accurate but is clinically useful. MR imaging provides information about associated injuries prior to surgery. Radiologic diagnostic methods for the assessment of ACL tears on MR images are well established; recent techniques such as dedicated cartilage imaging offer new information of use to clinicians about the sequelae of ACL injuries. PMID- 12528909 TI - Graft selection in anterior cruciate ligament reconstruction. AB - Selecting the appropriate graft for ACL reconstruction depends on numerous factors including surgeon philosophy and experience, tissue availability (affected by anatomical anomalies or prior injury or surgery), and patient activity level and desires. Although the patella tendon autograft has the widest experience in the literature, and is probably the most commonly used graft source, this must be tempered with the higher reported incidences of potential morbidity and pitfalls associated with its use. The hamstring tendons are gaining increasing popularity, mostly due to reduced harvest morbidity and improved soft tissue fixation techniques, and many recent studies in the literature report equal results to BTB ACL reconstruction with respect to functional outcome and patient satisfaction. On the other hand, many of these studies report higher degrees of instrument (KT-100) tested laxity for hamstring reconstruction, and some have reported lower returns to preinjury levels of activity. One question that remains to be addressed is how closely objectively measured laxity tests correlate with subjectively assessed outcomes and ability to return to high levels of competitive sports. Allograft use, which decreased in popularity during the 1990s, appears to be undergoing a resurgence, with better sterilization processes and new graft sources (tibialis tendons), leading to increased availability and improved fixation techniques. The benefits of decreased surgical morbidity and easier rehabilitation must be weighed against the potential for greater failure of biologic incorporation, infection, and possibly slower return to activities. In our practice, for high-demand individuals (those playing cutting, pivoting, or jumping sports and skiing) BTB tends to be the graft of choice. For lower demand or older individuals, hamstring reconstructions will be performed. Allograft tissue will be used in older individuals (generally over 45 years old), those with signs of arthritis (and compelling evidence of instability), or those individuals who understand the pros and cons of allograft use fully and do not want their own tissue used. PMID- 12528910 TI - Anterior cruciate ligament graft fixation. AB - ACL reconstruction, when performed correctly, has become a reliable, reproducible surgical technique with a predictable outcome. Successful results can be achieved via anatomic reconstruction with rigid fixation of a strong graft. Fixation techniques continue to evolve, with biologic fixation of graft constructs gaining popularity over the past several years. The choice of fixation should be one that provides a firm construct where accelerated rehabilitation can be achieved on all isolated primary ACL reconstructions. The results should be reproducible and, most important, the surgeon should be comfortable and confident with the fixation device. Prior to selecting a particular fixation device, the surgeon should not only be aware of any potential problems noted on biomechanical testing but also have familiarity with the proper use of the device and any clinical follow-up studies in the literature. PMID- 12528911 TI - Tensioning of the anterior cruciate ligament graft. AB - A great deal in variability exists in the protocols for graft tensioning. Biologic studies have not been conclusive about the optimum tension for revascularization while minimizing stress relaxation. Clinically, the risk of undertensioning the graft, and thereby not correcting the laxity, must be balanced with the risk of overconstraining the knee, which may lead to pathologic stresses on the joint cartilage, graft failure, or infrapatellar contracture syndrome . Amis and Jakob reported at the European Society of Sports Traumatology Knee Surgery and Arthroscopy scientific workshop in 1998 that most surgeons chose to impose a graft tension of 20 N to 80 N at an average of 11 degrees flexion. Preloading in full or nearly full extension limits the risk of pathologic stresses that can occur at full extension when the graft is tensioned at 30 degrees, but increases the risk of laxity at that angle. For our flexion parameters, we recommend tensioning the graft at 20 degrees to 30 degrees unless the graft tightens or shortens 5 mm or more with knee extension. If shortening occurs, tensioning should be performed in extension. The magnitude of graft tensioning should be tissue specific. For bone-patellar tendon-bone autograft or allograft, 5 pounds of tension should be applied. For hamstring graft, we apply 15 pounds of tension with each bundle tensioned individually. PMID- 12528912 TI - Endoscopic anterior cruciate ligament reconstruction with patella tendon autograft. AB - The patella tendon is the most commonly used graft source for ACL reconstruction because of its biomechanical strength and stiffness, the availability of bone-to bone healing on either end, and the ability to firmly secure the graft within the tunnels. Consistently good results have been reported in the literature, with expectations to return to all activities at pre-injury levels of performance. We outline our technique for endoscopic ACL reconstruction using a patella tendon autograft. The technique is divided into the critical stages of pre-operative assessment, graft harvest, notch preparation, tunnel placement, graft passage, graft fixation, and rehabilitation. Methods for avoiding pitfalls and overcoming mishaps are described. PMID- 12528913 TI - Anatomic endoscopic anterior cruciate ligament reconstruction with patella tendon autograft. AB - Current tibial endoscopic ACL reconstruction techniques provide functional stability, but fall short of the ultimate goal of ACL reconstruction, to restore normal knee kinematics. Vertical graft placement results in restoration of normal anteroposterior stability with a negative Lachman exam, but may not produce a stable knee in rotation, noted by a positive pivot shift. The Clancy anatomic endoscopic ACL reconstruction technique utilizes flexible reamers to achieve anatomic graft placement to more closely reproduce normal knee function. The overall results of arthroscopic anatomic endoscopic ACL reconstruction are essentially the same as we have reported using our previous open and rear-entry, two-incision techniques for anatomic graft placement. The long-term benefits of a more physiologic single incision endoscopic ACL reconstruction are not yet determined; however, short-term results are encouraging. PMID- 12528914 TI - Anterior cruciate ligament reconstruction the two-incision technique. AB - Historically, the two-incision technique for anterior cruciate ligament (ACL) reconstruction was the standard of practice. It allows predictable, near-anatomic placement of the femoral tunnel and provides highly reproducible results with few complications. There are several major advantages of the two-incision technique over endoscopic methods. These include consistent femoral tunnel placement, elimination of concern for "blowing out the back wall," elimination of the problem of graft-tunnel mismatch, elimination of the problem of screw divergence, and ease of use for revision ACL reconstruction procedures. The angle of the ACL graft is also more anatomic, matching the angle of the native ACL. In this chapter we present our technique for reconstruction of the ACL using the two incision approach. Indications for its use are discussed. Advantages and disadvantages of the two-incision method compared with the all-endoscopic approach are reviewed. PMID- 12528915 TI - The Sivaganga eye survey: I. Blindness and cataract surgery. AB - PURPOSE: To assess the prevalence of visual acuity impairment, blindness, and cataract surgery among older adults in rural southern India. METHODS: Random selection of village- and urban-based clusters was used to identify a cross sectional sample of persons 50 years of age or older from the Sivaganga district of Tamil Nadu. Subjects in 25 selected clusters were enumerated through a door-to door survey and invited to examination sites for measurement of uncorrected, presenting, and best-corrected visual acuity and ocular examination in 1999. The principal cause was identified for eyes with presenting visual acuity worse than 6/18. Quality assurance monitoring of visual acuity measurements took place in five of the study clusters. RESULTS: A total of 5081 persons in 3517 households were enumerated, and 4642 (91.4%) were examined. Thirty-six percent presented with visual acuity worse than 6/18 in the better eye. The prevalence of blindness, based on visual acuity worse than 6/60 in both eyes, was 6.0% (95% confidence interval [CI]: 5.1% to 6.9%) with presenting vision, and 2.5% (95% CI: 1.8% to 3.1%) with best correction. Blindness with presenting visual acuity was associated with older age and illiteracy. Cataract was the principal cause of blindness in one or both eyes in 69.4% of those presenting blind, and uncorrected aphakia and other refractive error affected 35.6% in at least one eye. The prevalence of cataract surgery was 14.7% (95% CI: 13.0% to 16.4%); low surgical coverage among the cataract blind was associated with illiteracy. CONCLUSIONS: It appears that much has been done in the prevention of blindness in Sivaganga. Nevertheless, blindness remains an important public health problem, mainly because of cataract and refractive error. Prevention of blindness programs in the area should target these two causes, with special emphasis on the elderly and the illiterate. PMID- 12528916 TI - The Sivaganga eye survey: II. Outcomes of cataract surgery. AB - PURPOSE: To assess the clinical outcomes of cataract surgery in rural southern India. METHODS: Cluster sampling was used in randomly selecting a cross-sectional sample of persons 50 years of age or older for visual acuity measurement, refraction, and ocular examination in 1999. Subjects were queried as to the date and place of surgery for each cataract-operated eye. Surgical complications were noted and the principal cause of reduced vision was identified for eyes with presenting visual acuity worse than 6/18. RESULTS: Within the cataract-operated sample of 682 persons, 13.8% had presenting visual acuity worse than 6/60 in both eyes, 25.2% better than or equal to 6/18 in both eyes, and 37.8% were bilaterally operated on. For aphakic eyes, 50.5% presented with visual acuity better than or equal to 6/18; 82.6% with best-correction. For pseudophakic eyes, the corresponding percentages were 78.0% and 94.5%. Over one-third of all eyes were pseudophakic, and nearly three-fourths had been operated on in non-governmental facilities. Uncorrected aphakia and other refractive error were the main causes of vision impairment. In multiple logistic regression modeling, poor presenting visual acuity in aphakic eyes was associated with illiteracy, rural residence, and surgery in government facilities; gender and time period of surgery were not predictors of vision outcomes. None of these variables were associated with best corrected outcomes in aphakic eyes, nor with presenting and best-corrected outcomes in pseudophakic eyes. CONCLUSIONS: Visual acuity outcomes in pseudophakic eyes were good. More attention must be given to needless vision impairment among the cataract-operated because of inadequate aphakic correction, especially among those operated on in government facilities, the illiterate, and those living in rural villages. PMID- 12528917 TI - The prevalence of Behcet's disease above the age of 10 years. The results of a pilot study conducted at the Park Primary Health Care Center in Ankara, Turkey. AB - PURPOSE: The aim of this study was to determine the prevalence of Behcet's disease above the age of 10 years by means of a population-based study. METHODS: The epidemiological investigation (cross-sectional study) was made between May 1997 and May 1998 at the Park Primary Health Care Center, which is one of the education and research divisions of the Department of Public Health, Faculty of Medicine, Ankara University. The research aimed to cover all 17,256 (49.2% male, 50.8% female) inhabitants over 10 years of age living in this area. The screening team first surveyed and selected patients with recurrent aphthous stomatitis. These patients were further examined, free of charge, in the Preventive Ophthalmology Unit of the Public Health Center, at Ibni Sina Hospital's Behcet Center or in other clinics if necessary. In this study the International Study Group For Behcet's disease Criteria were used. RESULTS: As the final result of the screening, 11 female and 5 male patients with Behcet's disease were found (female/male = 2.2). These patients represented 9 already known and 7 newly diagnosed cases of Behcet's disease. The prevalence of Behcet's disease over 10 years of age is 0.11%. CONCLUSION: The existing regional prevalance studies conducted in Turkey have indicated that the real number of Behcet's patients in our country is markedly higher than the number of registered patients. Therefore the National Behcet's Disease Commity and Surveillance System was founded by our research group in December 1999. PMID- 12528918 TI - Bias in self-reported family history and relationship to glaucoma: the Blue Mountains Eye Study. AB - PURPOSE: To examine bias in the relationship between self-reported family history of glaucoma and its relationship to the prevalence of glaucoma and ocular hypertension. METHODS: In a cross-sectional population-based study of 3654 Australians aged 49-97, participants were asked whether any first-degree relatives had been diagnosed with glaucoma. Open-angle glaucoma was diagnosed from matching optic disc and typical visual field changes, after gonioscopy. Ocular hypertension (OH) was diagnosed from elevated intraocular pressure (IOP) in subjects without glaucoma. RESULTS: Glaucoma was present in 3.0% and ocular hypertension in 5.2% of subjects. A parent or sibling was reported to have glaucoma by 8.6%, including 10.5% of women and 5.9% of men. A positive family history was reported more frequently in parents (6.4%) than siblings (2.6%). Glaucoma was reported more frequently to affect mothers (5.0%) and sisters (1.6%) than fathers (1.5%) and brothers (1.2%). A first-degree family history was given by 15.7% of subjects with glaucoma compared to 8.3% of controls, odds ratio (OR) 3.2 (95% CI 1.8-5.6), after adjusting for glaucoma risk factors, including IOP. The association had a similar magnitude for a family history in parents and siblings. Although recall bias was evident from the finding of increased odds (OR 4.2) among previously diagnosed cases, the relationship with family history also persisted in newly-diagnosed cases (OR 2.4). A slightly stronger relationship was found between OH and glaucoma family history, OR 3.9 (95% CI 2.6-5.7), after adjusting for confounders, but was also strongly influenced by recall bias. CONCLUSIONS: Although a positive family history of glaucoma may help to identify those at risk, it is subject to recall, selection and survival bias as well as community under-diagnosis of glaucoma and will most likely substantially underestimate the genetic influence. PMID- 12528919 TI - Comment on visual acuity. PMID- 12528920 TI - Involvement by young drivers in fatal alcohol-related motor-vehicle crashes- United States, 1982-2001. AB - Motor-vehicle crashes are the leading cause of death in the United States for persons aged 16-24 years, and a substantial proportion of these crashes are alcohol-related. Alcohol-impaired driving is highest among persons aged 21-24 years, and the percentage of fatal crashes that are alcohol-related is highest for this age group. However, alcohol-related crashes are a serious problem even for the youngest drivers. Not only are drivers aged <21 years more likely than older drivers to be involved in fatal crashes, but their added risk for fatal crash involvement increases more sharply at all levels of alcohol use. To characterize the rate of alcohol-related fatal crashes among young drivers, CDC analyzed unpublished data from the Fatality Analysis Reporting System (FARS), a national database of information on fatal crashes. The findings indicate that the largest decrease in alcohol-related fatal crashes during 1982-2001 was among drivers aged <21 years, who have been the target of several interventions to reduce alcohol-impaired driving. Public health and traffic safety professionals should ensure that communities implement comprehensive and effective strategies to reduce alcohol-impaired driving. PMID- 12528921 TI - State-specific trends in U.S. live births to women born outside the 50 states and the District of Columbia--United States, 1990 and 2000. AB - Persons born outside the 50 states and the District of Columbia (DC) comprised an estimated 11.1% (31.1 million) of the U.S. population in 2000, and approximately one fifth of all U.S. births in 2000 were to women in this population. Racial/ethnic disparities in U.S. health outcomes are of public health concern, and the increasing cultural and linguistic diversity of the U.S. population poses challenges to the delivery of maternal and child health services. This report presents state-specific comparisons of live births in 1990 and 2000 to women born outside the 50 states and DC and compares maternal characteristics and live-birth outcomes for these women with those of state-born mothers (i.e., women born inside the 50 states and DC). The findings indicate that women born outside the 50 states and DC had better birth outcomes than their state-born racial/ethnic counterparts. However, a larger percentage of these women began prenatal care later and had other problems accessing health care, which might reflect economic, cultural, and language barriers. The U.S. public health system and maternal healthcare providers should understand and address the health needs of an increasingly diverse population. PMID- 12528922 TI - Update: Influenza activity--United States, 2002-03 [corrected] season. AB - This report summarizes influenza activity in the United States during the weeks ending October 5-November 23, 2002. Influenza activity was low during this period but is expected to increase during the coming weeks. Vaccine supplies are plentiful this year, and vaccination should continue throughout December and the remainder of the influenza season, particularly among persons at high risk for complications from influenza. PMID- 12528923 TI - The importance of Doppler studies in asymptomatic intracranial and extracranial arterial disease. AB - Knowledge of intracranial and carotid disease in the symptomatic and especially the asymptomatic high risk population may be useful for evaluating future treatment modalities. A group of 204 symptomatic patients and 105 asymptomatic elderly patients at high risk for stroke were tested by carotid duplex ultrasound and transcranial doppler for the presence of carotid and intracranial stenosis. Quantitative measurements of the stenosis were made directly from the hard copy of the carotid duplex and the transcranial doppler. Of the 204 symptomatic patients 168 (83%) had some degree of stenosis: 84 of the 204 (41%) in the intracranial circulation only, 59 (29%) in the internal carotid only, and 26 (13%) in both sets of vessels. Of the asymptomatic patients 85 (81%) had some degree of stenosis; 31 of 105 (30%) in the intracranial circulation only, 35 (33%) in the internal carotid only, and 19 (18%) in both sets of vessels. Statistical analysis did not reveal significant differences between the two groups. The large percentage of intracranial disease in the symptomatic as well as the asymptomatic population at high risk for stroke require further confirmation by good duplex studies of the intracranial circulation. This is important in order to create coherent treatment protocols. PMID- 12528924 TI - Arteriovenous shunting blood flow is intravitally observed in the stomach after thermal injury in rats. AB - Microcirculatory disturbance of the gastric wall is a crucial factor in the development of gastric mucosal lesions induced by Helicobacter pylori, anti inflammatory drugs and stress. Opening of the arteriovenous shunting channel after thermal injury is one of the possible mechanisms to reduce mucosal blood flow. However, no in vivo observation of arteriovenous shunting blood flow in the stomach has been reported. To assess gastric microcirculatory disturbance, especially arteriovenous shunting blood flow after thermal injury by in vivo microsciopy, male Wistar rats were anesthetized and thermal injury was inflicted on the back skin. Gastric microvascular images were observed by in vivo microscopy. Rolling of leukocytes labelled with carboxyfluorescein diacetate, succinimidyl ester were counted and blood flow dynamics were observed by flow of a micro dye, monastral blue B (MBB). The endothelial damage was assessed by deposits of MBB 5 min after the administration. Arteriovenous shunting blood flow is difficult to detect by normal methods, but it could be observed by How of MBB after thermal injury. Statistical analysis showed a significant difference in the ratio of arteriovenous shunting blood flow detection between the control (no injury) (0%; n = 15) and thermal injury (5 hrs after thermal injury) (28.6%; n = 14) groups. In the thermal injury group, the percentage of rolling leukocytes and the area of monastral blue B deposits increased, and the venular walls tended to be irregular. The total length of erosion increased time-dependently after thermal injury, and the length at 5 hrs was approximately 2 times larger than that at 2 hrs. Arteriovenous shunting blood flow is intravitally observed after thermal injury. A-V shunting blood flow can be a cause of mucosal hypoperfusion. It is suggested that the microcirculatory disturbance seen 5 hrs after thermal injury is contributed to the final step of erosion formation. PMID- 12528925 TI - In vivo measurement of superoxide in the cerebral cortex during anoxia reoxygenation and ischemia-reperfusion. AB - Abstract. The exact time profile of superoxide generation during anoxia reoxygenation and ischemia-reperfusion was assessed in the feline cerebral cortex in vivo using a chemiluminescence technique with a probe specific for superoxide, 2-methyl-6-[p-methoxyphenyl]-3,7-dihydroimidazo[1,2-alpha]pyrazin-3-one (MCLA). MCLA solution was superfused on the cortex throughout the protocol, and MCLA chemiluminescence was measured using a newly developed photon counting system. Reflectance at 398 nm was simultaneously measured to compensate for hemodynamic artifacts resulting from cerebral blood volume changes. In 19 animals, a 90 second anoxia was induced by the inhalation of 100% nitrogen followed by a 40 minute reoxygenation. The chemiluminescence decreased during the period of anoxia (p < 0.01) and then exceeded the baseline level at 15 and 20 minutes after reoxygenation (p < 0.05). In six animals, superoxide dismutase (SOD) was continuously superfused and anoxia-reoxygenation was performed in the same manner. The chemiluminescence decreased during the period of anoxia (p < 0.05) but did not exceed the baseline level during the reoxygenation period, indicating that an increase in superoxide production was the main cause of the chemiluminescence increase. In eight animals, a 15-minute forebrain ischemia was induced by the occlusion of the bilateral common carotid arteries with systemic hypotension (systolic blood pressure less than 50 mmHg) followed by a 30-minute reperfusion. The chemiluminescence decreased during the period of ischemia (p < 0.01) and then increased at 20 and 25 minutes after reperfusion (p < 0.05). These results indicate that superoxide generation decreases during anoxia and ischemia and then increases within 20 minutes after reoxygenation or reperfusion. PMID- 12528926 TI - Is cardiomyopathy an autoimmune disease? AB - Idiopathic dilated cardiomyopathy (DCM) is one of the leading causes of severe heart failure and the most common cause of heart transplantation due to its ventricular dilatations and contractile dysfuntions. Twenty percent of DCM is in the familiar form and the rest is sporadic. The clinical impact of DCM is far greater than its position in epidemiological terms. Despite recent improvements in therapy, both incidence and mortality are still very high. The main problem is its heterogeneous etiology. So far, three factors have been identified to be potentially important: enteroviral infection, immune mechanism and genetic factors. During the last 10 years there have been many investigations showing distinct autoantibodies or other immune factors in heterogeneous subsets of DCM which have contributed supportive and confounding evidence to the hypothesis that multiple autoimmune mechanisms are involved in DCM. Accumulated evidence hitherto demonstrated a variety of circulating autoantibodies in the sera of patients with DCM including antireceptor autoantibodies, myosin and ADP/ATP translocator protein, etc. Data available from both in-vitro and in-vivo studies of anti receptor autoantibodies as well as from other autoantibodies and autoreactive lymphocytes demonstrated clearly that a subgroup of DCM is autoimmunity-mediated. This is understandable because DCM is heterogeneous, implying that different subgroups of DCM may have different pathogeneses. It may be practical in the future to separate "autoimmune cardiomyopathy" from other "idiopathic" DCM. PMID- 12528927 TI - Introductory remark of Dr. Rokuzo Kobayashi's achievements. PMID- 12528928 TI - Special remark: a portrait of Dr. Rokuzo Kobayashi. PMID- 12528929 TI - Helicobacter pylori type IV secretion, host cell signalling and vaccine development. AB - Helicobacter pylori is a bacterial pathogen specialised to colonise and persist the gastric mucosa and to cause severe gastroduodenal disease. A major disease associated bacterial component is a type IV secretion system (TFSS) encoded by the cytotoxin-associated genes pathogenicity island (cagPAI). Among the multiple responses in H. pylori-infected epithelial cells, the induction of proinflammatory cytokines and chemokines, cell spreading and motility associated with the "hum mingbird" phenotype appear strictly dependent on the functional transporter complex in the cagPAI. H. pylori is also capable of occasionally entering epithelial cells and manipulates the host immune system for immune evasion. Attached bacteria actively translocate the CagA protein into epithelial cells by a TFSS-dependent process and translocated CagA undergoes tyrosine phosphorylation in the carboxy terminal EPIYA sequence repeat motif (Y-972) by kinases of the Src family. Furthermore, we have identified a novel TFSS in H. pylori involved in horizontal DNA-transfer. Host cell signalling events and cellular phenotypes provoked by the cagPAI, the investigation of mechanisms related to gastric cancer as well as the development of a Salmonella based live recombinant vaccine are in the focus of additional departmental activities. PMID- 12528930 TI - Growth cycle of Helicobacter pylori in gastric mucous layer. AB - Helicobacter pylori bacterium is characterized by its strong urease activity. Our studies on the role of H. pylori urease revealed; (i) it is essential for colonization, (ii) exogenous urea is required for acid resistance, (iii) the bacteria have the ability to move toward urea and sodium bicarbonate, (iv) urea hydrolysis accelerates chemotactic locomotion, and (v) decay of urease mRNA to accomplish the active center is pH-regulated; i.e., the mRNA is stabilized and destabilized under acidic and neutral conditions, respectively. Based on the above results, I propose the growth cycle of H. pylori in gastric mucous layer. H. pylori bacteria proliferate on the epithelial cell surface by utilizing nutrients derived from degraded cells. Proliferated bacteria leave the cell surface to pH-variable region where they encounter strong acid. Urease is activated with simultaneous opening of UreI channel so that urea is hydrolyzed to neutralize acid. Chemotaxis of H. pylori toward urea and sodium bicarbonate that are abundant on the cell surface is accelerated by urea hydrolysis so that the bacteria go back to the cell surface for the next round of proliferation. This growth cycle may allow the bacteria to infect persistently in the stomach. PMID- 12528931 TI - Protein tyrosine phosphatase beta, a receptor for Helicobacter pylori vacA toxin. AB - Helicobacter pylori is the leading bacterial cause of food-borne illness worldwide and plays a major role in the development of chronic gastritis, peptic ulcer, and gastric cancer. Strains isolated from patients contain the cagA gene (cytotoxin-associated gene A) and produce the vacuolating cytotoxin VacA. VacA binding to specific high-affinity cell surface receptors was shown by using indirect immunofluorescence and flow cytometry; high-affinity toxin binding was necessary for cell intoxication. A 250-kDa receptor protein tyrosine phosphatase (RPTP) beta served as a receptor for VacA on AZ-521 cells. The overexpression of RPTP beta conferred VacA sensitivity on BHK-21 cells transfected with the RPTP beta cDNA, consistent with RPTP gamma acting as a receptor for VacA. Increased binding of acid- or alkali-activated VacA to RPTP gamma may alter its activity and possibly accelerate or inhibit dephosphorylation of tyrosine on cytosolic proteins. Understanding the pathological responses of wild type and RPTP gamma deficient animal models may well provide valuable information regarding the mechanism of VacA toxicity. PMID- 12528932 TI - Immune response to heat shock protein of Helicobacter pylori--a candidate as a vaccine component. AB - . The reactive epitope of heat shock protein 60 (HSP60) of Helicobacter pylori to its monoclonal antibody (H9) was determined, and its synthesized peptide designated pH9 was used for ELISA. The patients with H. pylori infection had significantly lower titers of pH9 antibody than did uninfected patients. In C57BL/6 mice immunized intraperitoneally with the pH9 peptide with Freund's complete adjuvant (FCA), the number of H. pylori organisms colonizing the stomach was significantly lower than that in mice immunized with FCA only. These results suggest that HSP60 of H. pylori is effective in protection against H. pylori infection and might be a good candidate as a vaccine component. PMID- 12528933 TI - Deregulation of SHP-2 tyrosine phosphatase by the Helicobacter pylori virulence factor CagA. AB - Helicobacter pylori (H. pylori) is estimated to infect about half of the world population. It causes gastric diseases ranging from gastritis to cancer and has been classified as a class I carcinogen by WHO. However, little is known about the molecular mechanisms by which H. pylori induces pathogenesis. CagA is the product of the cagA gene carried among virulent H. pylori strains and is associated with severe clinical outcomes, most notably gastric carcinoma. CagA is injected from the attached H. pylori into gastric epithelial cells and undergoes tyrosine phosphorylation. We found that wild-type, but not phosphorylation resistant CagA, is capable of inducing a growth factor-like morphological change, termed the hummingbird phenotype, in cells. Furthermore, CagA specifically binds the SH2-containing protein tyrosine phosphatase SHP-2 in a tyrosine phosphorylation-dependent manner and stimulates phosphatase activity. Disruption of the CagA-SHP-2 complex abolishes the CagA-dependent morphological change. Conversely, constitutively active SHP-2 is capable of inducing a CagA-like morphological change when it is plasma membrane-targeted. Our results show that CagA perturbs cellular functions by deregulating SHP-2 phosphatase after translocation from H. pylori into gastric epithelial cells. Given the positive regulatory roles of SHP-2 in both cell proliferation and cell movement, the CagA SHP-2 interaction may play an important role in the oncogenic transformation that is a hallmark of cagA+ H. pylori infection. PMID- 12528934 TI - Gastric spirochaetes: 100 years of discovery before and after Kobayashi. AB - The discovery of Helicobacter pylori, by Warren and Marshall in 1982, was preceded by nearly 100 years of inconspicuous publications relating to spiral bacteria, achlorhydria, gastritis, gastric urease, and antimicrobial therapy for ulcers. Japanese investigators, notably Kasai and Kobayashi, should be acknowledged for their pioneering work showing that spiral bacteria could infect many animals, could cause haemorrhagic erosions, and would be effectively cured with various antimicrobials. PMID- 12528935 TI - Special remarks to Rokuzo Kobayashi Memorial Symposium on Helicobacter pylori. PMID- 12528936 TI - Gastric mucosal response to Helicobacter pylori. AB - Since Marshall's discovery before 20 years, Helicobacter pylori (H. pylori) infection is reportedly to be associated with a variety of clinical outcomes including peptic ulcer disease and gastric cancer. The first step of the H. pylori colonization might be its adhesion to the surface epithelial cells, which evokes gastric inflammatory events initiated by neutrophil recruitment from the microcirculation. Mongolian gerbil is one of the suitable animal models for H. pylori infection, which exerts gastric ulcer and cancer with its bacterial infection. In H. pylori-colonized gerbils, extensive levels of microvascular leukocyte adhesion and migration into the parenchymal side and significant levels of inflammatory cell infiltration are encountered. Bacterial urease not only neutralizes gastric luminal acid, but also plays as an adhesion factor to the surface epithelium. Recently, such an adhesion to the epithelium is reported to be important for bacterial type IV secretory system, which intermediates Cag A injection into the epithelial cells. Then, multiple chemokine and cytokine networks are activated and mucosal inflammatory lesion formation would be completed. In the long-term colonization of H. pylori, gastric mucosal cell turnover would be modified due to persistent inflammation and then such deregulation of cell turnover might link to the precancerous lesion formation. PMID- 12528937 TI - Effect of Helicobacter pylori-mediated inflammation on nonsteroidal anti inflammatory drugs-induced gastric mucosal injury. AB - Helicobacter pylori (H. pylori) infection and nonsteroidal anti-inflammatory drugs (NSAIDs) are two major causes of gastric ulceration, but relation between H. pylori infection and use of these drugs in gastric mucosal injury is controversial. Neutrophils have been implicated in the pathogenesis of gastric mucosal damage induced by H. pylori or NSAIDs. H. pylori itself and H. pylori extract induce neutrophil activation, such as superoxide production, expression of adhesion molecule (CD11b/CD18) and transendothelial migration, capillary plugging. Several clinical and experimental studies demonstrated that the degree of H. pylori-induced gastric mucosal injury was closely correlated with the extents of H. pylori infection and of neutrophil infiltration, suggesting implication in extravascular neutrophils for H. pylori-induced gastric mucosal injury. On the other hand, aspirin promotes neutrophil-endothelial adhesive interactions via increasing CD11b/CD18, but not neutrophil migration to extravascular space. In addition, our recent in vivo study suggests that neutrophils adhering to the blood vessels, but not neutrophils migrating to the interstitium, are implicated in aspirin-induced gastric mucosal injury. Recently, we found that administration of aspirin to gerbils three weeks after H. pylori inoculation produced severe gastric mucosal injury via marked infiltration of neutrophils. In this animal model, pretreatment with anti-neutrophil serum, elastase inhibitor or scavengers of reactive oxygen species remarkably inhibited gastric mucosal injury. These results suggest that H. pylori infection potentiates aspirin-induced gastric mucosal injury by mechanisms that include accumulation of activated neutrophils. PMID- 12528938 TI - Cytokine expressions and H. pylori-associated gastric mucosal lesion. AB - To determine the role of host genetic factors in Helicobacter pylori infection, we examined the relation between gastroduodenal diseases and IL-1B polymorphisms in patients with H. pylori infection. In addition, we also compared gastric mucosal cytokine levels in those patients. We confirmed the findings that the IL 1B-31 C-to-T base transition was inverted in association with the -511 T-to-C base transition. There was no relation regarding to IL-1B polymorphisms and clinical outcomes. The gastric mucosal IL-1B level of the body of the stomach but not the antrum was significantly different among IL-1B genotypes. Furthermore, the IL-8 levels in the body were also higher in IL-1B-511C/C/ IL-1B-31TT than H. pylori negative patients. These findings suggested that IL-1B polymorphisms enhance not only IL-1-B production but also IL-8 production in the gastric body and may play an important role in the development of atrophic gastritis. PMID- 12528939 TI - How does H. pylori infection cause gastric cancer? AB - H. pylori is now recognised to be an important co-factor in the aetiology of non cardia gastric cancer of both the diffuse and intestinal histological type. The latter type develops via a complex multistage and multifactorial process. The fist stage involves progression from superficial gastritis to atrophic pangastritis with intestinal metaplasia and associated hypochlorhydria. This gastric phenotype may then progress to dysplasia and cancer. Many co-factors are involved in this progression including the strain of H. pylori, host genetic factors, such as interleukin-1 polymorphisms and gender, plus environmental factors such as smoking and diet. Intestinal colonisation with helminthic infection may retard the progression by altering the immune and inflammatory response to H. pylori and colonisation of the achlorhydric stomach with nitrosating bacteria may promote progression to cancer. H. pylori appears to be an obligatory co-factor in the aetiology of most gastric cancers. Consequently, prevention of the infection or its eradication in early life should reduce the incidence of this common and usually fatal tumour. PMID- 12528940 TI - Molecular basis for development of gastric cancer--in vivo and in vitro studies. PMID- 12528941 TI - H. pylori infection and the development of gastric cancer. AB - BACKGROUND: Recently, many study have shown that Helicobacter pylori infection is crucial in development of atrophic gastritis, which is closely associated with gastric cancer. We conducted a long-term endoscopic prospective follow-up study to investigate the development of gastric cancer in H. pylori-positive and negative patients. METHODS: 1,603 patients who underwent endoscopy and were assessed as to the presence of H. pylori infection by histology, rapid urease test and serologic test between April 1990 and March 1993 were entered. We prospectively studied 1246 subjects with and 280 subjects without H. pylori infection for a mean follow-up of 7.8 years (range 1-10.6 years). RESULTS: Gastric cancer of both the intestinal and diffuse type developed in 36 (2.9%) infected patients but in none of the uninfected patients during follow-up. There was an increased risk for gastric cancer in infected patients with severe gastric atrophy and corpus predominant gastritis and intestinal metaplasia. Gastric cancer was detected in 21 (4.7%) of the patients with non ulcer dyspepsia, in 10 (3.4%) of those with gastric ulcer and in 5 (2.2%) of those with gastric hyperplastic polyp, at enrollment. No gastric cancer was detected in duodenal ulcer patients. CONCLUSION: These results suggest that the development of both types of gastric cancer is caused by H. pylori-associated gastritis, and the risk for development of gastric cancer in H. pylori-negative subjects is extremely low. Subjects having H. pylori-positive gastric mucosa with severe atrophy and/or corpus gastritis may be at particularly high risk for gastric cancer. PMID- 12528942 TI - Helicobacter pylori and gastric carcinoma--from the view point of animal model. AB - Many epidemiological studies have shown a strong association between chronic Helicobacter pylori infection and subsequent development of gastric carcinoma in humans. To confirm this link more clearly, it is necessary to use this bacterium in experimental studies to develop gastric carcinoma in suitable experimental animals. Persistent H. pylori infection has recently been achieved in the Japanese Monkeys and Mongolian gerbil models, with results demonstrating that the sequential histopathological changes in the gastric mucosa are closely mimic the gastric mucosal changes caused by H. pylori infection in humans. Gastric mucosa infected with H. pylori exhibited significantly higher gastritis score, reduction in glandular height, increase in the number of Ki-67 positive cells and over expression of p53 protein and p53 gene mutation in the Japanese Monkey Model. In the Mongolian gerbil model, H. pylori infection enhances gastric carcinogenesis in combination with known carcinogens such as MNU and MNNG, and also demonstrated that H. pylori infection alone can result in the development of gastric carcinoma. However, diagnostic criteria of gastric carcinoma in animal models remain in the great discussion. These important results provide a starting point for further studies to clarify the mechanism of gastric carcinogenesis as a result of H. pylori infection and assist the planning of eradication therapy to prevent gastric carcinoma. PMID- 12528943 TI - To stay sharp, use a dull point. PMID- 12528944 TI - Decision making for preventive procedure. PMID- 12528945 TI - At-risk children of teen mothers. PMID- 12528946 TI - Caries-risk factors for Hispanic children affected by early childhood caries. AB - PURPOSE: The purpose of this study was to examine factors associated with early childhood caries (ECC) and to develop a profile of risk factors for Hispanic families affected by this condition. METHODS: Sixty Hispanic families with at least 1 child affected by ECC and 60 Hispanic families without any affected children were included in the study. Data was collected using a parent-completed questionnaire which gathered information about family demographics, dental care and hygiene practices, and feeding practices of the children. RESULTS: Parents in families without ECC were significantly more likely to have visited the dentist recently and the children were less likely to sleep while feeding compared to families with an ECC-affected child. Within families affected by ECC, siblings with ECC were significantly more likely to use the bottle and to sleep while feeding compared with their siblings without ECC. Fifty-five percent of the subject families had more than 1 child affected by ECC. However, odds ratios did not indicate an increased risk of ECC among younger siblings in the presence of affected older siblings. CONCLUSIONS: The current study, while supporting the long-held belief that feeding practices influence the susceptibility and course of ECC, demonstrates the role of other nonfeeding practices. PMID- 12528947 TI - Current understanding of the epidemiology mechanisms, and prevention of dental caries in preschool children. AB - The seriousness and societal costs of dental caries in preschool children are enormous. National data shows that caries is highly prevalent in poor and near poor US preschool children, yet this disease is infrequently treated. The etiology includes elevated colonization levels of mutans streptococci, high frequency sugar consumption, and developmental defects on primary teeth. A necessary first step in preventing dental caries in preschool children is evaluating the child's caries risk factors that include socioeconomic status, previous carious experience, presence of white spot lesions, presence of visible plaque, perceived risk by dental professionals, and microbiologic testing for the presence or quantity of mutans streptococci. Based on this knowledge, different preventive strategies, as well as different intensities of preventive therapies, can be employed. Caries preventive strategies in preschool children include diet modifications to reduce high frequency sugar consumption, supervised tooth brushing with fluoridated dentifrice, systemic fluoride supplements to children living in a nonfluoridated area that are at risk for caries, professional topical fluoride with fluoride varnish, and sealants for primary molars. PMID- 12528948 TI - Effects of different pacifiers on the primary dentition and oral myofunctional strutures of preschool children. AB - PURPOSE: The aim of this study was to evaluate the characteristics of the dental arches and some oral myofunctional structures in 36- to 60-month-old children who sucked a pacifier or did not have this habit. METHODS: Sixty-one children were divided into 3 groups: (1) those who never sucked a pacifier, (2) those who exclusively sucked a physiological pacifier, and (3) those who exclusively sucked a conventional one. A clinical examination was performed on the children to observe the relationship between the arches and their width, as well as the following oral myofunctional structures: lips, tongue, cheeks, and hard palate. RESULTS: Statistical analysis showed that: (1) the use of both ytpes of pacifiers led to anterior open bite (prevalence of 50% in both groups; P = . 001), (2) posterior crossbite was present only on children who had a pacifier-sucking habit, (3) the mean oveqrjt was greater on children who sucked physiological (3.6 mm) or conventional (3.7 mm) pacifiers when compared to those with no sucking habits (1.3 mm; P = .001), (4) intercanine distance of the upper arch was significantly smalelr on children who sucked pacifiers (29.6 mm in the physiological group and 29.2 mm in the conventional pacifier group) than those who did not (31.2 mm), and (5) the children who never sucked on a pacifier showed a higher prevalence of normality of cheek mobility (74%; P = .022) and hard palate shape (78%; P = .042). CONCLUSIONS: Children who sucked pacifiers, both conventional and physiological ones, showed higher prevalence of alterations in the relationship of the dental arches and orla myofunctional structures, when compared to those who never sucked a pacifier. PMID- 12528949 TI - The distal shoe space maintainer chairside fabrication and clinical performance. AB - The chairside-fabricated distal shoe appliance, with a stainless steel crown as the retainer, is an efficacious and cost-effective appliance for guiding the unerupted permanent first molar into position after premature loss or extraction of the second primary molar. The fabrication technique is illustrated in this case report and data is presented on the success rate of the appliance. PMID- 12528950 TI - Management of fused supernumerary teeth in children using guided tissue regeneration: long-term follow up of 2 cases. AB - Surgical separation of supernumerary teeth fused to permanent incisor teethhas typically given rise to residual post surgical periodontal defects, induding loss of attachment and deep periodontal pocketing with persistent inflammation. Other complications include devitalisation of the retained tooth section, ankylosis, external and replacement resorption. A unique technique of using guided tissue regeneration has been successfully employed to promote periodontal healing, after 2 cases of surgical removal of a supernumerary tooth fused to a permanent maxillary lateral incisor tooth. In the first case, a 2-stage guided tissue regeneration technique was completed with a nonresorbable Gor-Tex membrane, and was followed up after 9 years. The second case was completed using a resorbable Vicryl membrane, in a single-stage guided tissue regenerative technique; and was followed up after 5 years. PMID- 12528951 TI - Acute onset of facial nerve palsy associated with Lyme disease in a 6 year-old child. AB - Pediatric facial nerve palsy (FNP) can result from a variety of etiologies including Lyme disease, varicella, primary gingivostomatitis, herpes zoster oticus (Ramsay Hunt syndrome), coxsackievirus, trauma, otitis media, HIV, diseases causing tumors or demyelinations, compressions, and possibly Epstein Barr virus. Lyme disease has been implicated as the cause of over 50% of the FNPs in children. The paralysis of the facial nerve disturbs motor function to the muscles of facial expression and results in a flaccid appearance of the face (unilateral or bilateral). This case report derails undiagnosed Lyme disease presenting as a facial palsy in a 6 year, 5 month-old white female. The palsy was recognized and consultation with the child's physician prompted definitive diagnosis and treatment. A review of the literature and the implications of facial nerve palsy are discussed. PMID- 12528952 TI - Neurofibromatosis type 1: a clinicopathological study of the orofacial manifestations in 6 pediatric patients. AB - Neurofibromatosis type 1 (NF1) is a relatively frequent mucocutaneous syndrome, which is transmitted as an autosomal dominant trait or which may represent neomutation. It is characterized by a variety of clinical manifestations, including multiple neurofibromas that are associated with a high risk of sarcomatous transformation. The aim of this report was to elucidate the orofacial manifestations observed in 6 pediatric patients (between 4 and 15 years of age) diagnosed with NF1. Physical, clinical, radiological, histological, and immunohistochemical studies were performed. Orofacial lesions were observed in all studied patients, located either in the soft tissues (4 cases) or centrally in the jaws (2 cases). All cases showed facial asymmetry, one of them exhibiting marked facial hemihypertrophy. All cases with soft tissue involvement were plexiform neurofibromas, while the intraosseous cases were diagnosed as solitary neurofibromas. Knowledge of the variability of presentation of orofacial soft tissue and bone manifestations of NF1 in children is necessary for prompt diagnosis. PMID- 12528953 TI - Can salivary composition and high flow rate explain the low caries rate in children with familial dysautonomia? AB - PURPOSE: Extremely low caries rate and increased major salivary gland flow rare have been previously reported in children with familial dysautonomia (FD). The purpose of this study was to explore the possibility that, in addition to increased salivary flow, children with FD have alterations in their salivary components, which may suggest an explanauon to their low caries rate. METHODS: Whole unstimulated and stimulated saliva samples were collected from 13 children with FD who were found to be caries free, and from 28 age- and ethnic-matched healthy children, 15 caries-free children and 13 caries-affected children. The electrolyte and protein content of the unstimulated saliva and the microbial count and buffering capacity of the stimulated saliva were determined. RESULTS: Children with FD had the highest salivary flow rate and the lowest levels of mutans streptococci and lactobacilli, as well as the lowest concentration of chloride, magnesium, total protein and IgA. Healthy caries-affected children displayed the highest mutans streptococci and lactobacilli levels and lysozyme concentration, concomitantly with the lowest potassium and calcium concentrations. CONCLUSIONS: The results of this investigation suggest that the caries-free state in FD may be associated with high salivary flow rate, while in healthy children, low caries rate may be associated with high salivary calcium concentration. PMID- 12528954 TI - Microleakage of adhesive resin systems in the primary and permanent dentitions. AB - PURPOSE: The purpose of this study was to compare the in vitro microleakage of fourth-generation filled and unfilled adhesive resin systems with fifth generation filled and unfilled adhesive resin systems in both primary and permanent teeth. METHODS: Eighty extracted or exfoliated human noncarious teeth (40 primary and 40 permanent) were assigned to each of 8 groups. Groups 1, 3, 5 and 7 were primary teeth, and groups 2, 4, 6 and 8 contained permanent teeth. Groups 1 and 2 were bonded with Optibond Fl (Kerr), groups 3 and 4 with Scotchbond Multipurpose (3M), groups 5 and 6 with Optibond Solo Plus (Kerr) and groups 7 and 8 with Single Bond (3M). All teeth received a Class V cavity preparation, and the cavosurface margins were placed entirely in enamel. They were then restored with TPH Spectrum Shade Al (Dentsply Caulk). All teeth were thermocycled, stained with basic fuchsin, sectioned and viewed under the microscope. Measurements were recorded in absolute millimeters and relative grades as judged by 2 evaluators. RESULTS: No significant difference in microleakage was observed between fourth- and fifth-generation adhesive resin systems, whether filled or unfilled, or applied on primary or permanent teeth. Significant differences were found in the amount of microleakage at the gingival and occlusal surfaces in all groups. One-bottle, fifth-generation adhesive resin systems permit easier application with the same effectives as the 2-bottle, fourth-generation systems. CONCLUSIONS: One-bottle, fifth-generation adhesive resin systems permit easier application with the same effectiveness as the 2 bottle, fourth-gerenration systems. PMID- 12528955 TI - Pulpal blood flow in vital and nonvital young permanent teeth measured by transmitted-light photoplethysmography: a pilot study. AB - PURPOSE: The purpose of this study was to record transmitted-light photoplethysmography (TLP) from young permanent teeth and examine its applicability to pulp vitality assessment. METHODS: Twenty-six healthy and 7 nonvital upper maxillary incisors in 17 children (ages 7 years, 3 months to 14 years, 8 months) were examined. Recordings of TLP were made with and without opaque black rubber dam application in a darkened room. Finger photoplethysmography (FPP) of the participants was simultaneously recorded. RESULTS: (1) Pulse waves synchronous with FPP were recorded from all healthy teeth, whereas no pulse signals were recognized in nonvital teeth. (2) The signal amplitude in the TLP was not significantly affected by the opaque dam application. (3) In the healthy teeth, there was a significantly negative correlation between the TLP pulse amplitude and the age of the subjects. CONCLUSIONS: TLP can detect pulpal blood flow in young permanent teeth. This technique is considered to be applicable in the assessment of pulp vitality. PMID- 12528957 TI - On the edge of faith. PMID- 12528956 TI - Chronic diseases and cancer. PMID- 12528958 TI - National stroke audit. PMID- 12528959 TI - Semi-creative tension? PMID- 12528960 TI - Medically unexplained symptoms and syndromes. PMID- 12528961 TI - Current approaches in chronic pain. PMID- 12528962 TI - Self-poisoning and self-injury in adults. PMID- 12528963 TI - Management of substance misuse problems in the general hospital. PMID- 12528964 TI - Neuropsychiatry. PMID- 12528965 TI - Consent with understanding: a movement towards informed decisions. AB - The development of consent processes over the last decade is reviewed. The need for an understanding of information is recognised, although the legal definition of competence does not require actual understanding, but rather capacity to understand. The need to move responsibility for procedures towards patients is discussed in terms of an informed decision about choices rather than an informed consent to a single option. PMID- 12528966 TI - Are preventive drugs preventive enough? A study of patients' expectation of benefit from preventive drugs. AB - OBJECTIVES: The study aimed to find the threshold of benefit for a hypothetical cholesterol-lowering drug below which the subject would not be prepared to take the drug. We also looked at whether proximity to the target event (myocardial infarction) and the subjects' views on drug taking affected this threshold. DESIGN: We studied 307 subjects using a written questionnaire and interview. Group 1 (102 subjects) had just been discharged from the coronary care unit. Group 2 (105 subjects) were taking cardio-protective drugs but had no recent history of myocardial infarction. Group 3 (100 subjects) had no history of myocardial infarction and were taking no cardio-protective drugs. RESULTS: Median values for the threshold of benefit below which the subject would not take the preventive drug were 20%, 20%, and 30% absolute risk reduction for Groups 1, 2 and 3 respectively. Median values for expectation of average prolongation of life were 12, 12 and 18 months respectively. Only 27% of subjects would take a drug offering 5% or less absolute risk reduction over five years. Subjects' views on medicinal drug taking in general and proximity to the target event were predictors of the acceptance of preventive drugs. Eighty percent of subjects wished to be told the numerical benefit of a preventive drug before starting on it. CONCLUSION: For the majority, the expectation of benefit from a preventive drug is higher than the actual benefit provided by current drug strategies. There is a tension between the patient's right to know about the chance of benefiting from a preventive drug and the likely reduction in uptake if they are so informed. PMID- 12528967 TI - Research ethics committees--time for change? AB - Research ethics committees have developed over the last 30 years in the UK. As their roles and responsibilities have grown, so too has the complexity of their work. This poses increasing problems which cannot be solved within the current structure. This article suggests that the present large number of volunteer committees be replaced by a much smaller number of fully professional bodies in order to increase speed, efficiency and consistency of working. PMID- 12528968 TI - Diastolic heart failure in older people--myth or lost tribe? AB - Left ventricular diastolic dysfunction occurs due to a variable combination of abnormal myocardial relaxation and reduced ventricular compliance. The diagnosis of diastolic congestive heart failure is controversial. Some studies suggest that up to one-third of older people with symptomatic congestive heart failure (CHF) have echocardiograph evidence of diastolic dysfunction. Other authors have suggested the comorbid diseases often found in persons with suspected diastolic CHF explain the patient's symptoms and hence diastolic CHF is a misdiagnosis in many cases. Many of the characteristic echo features of diastolic dysfunction occur in normal ageing hearts. Unlike in systolic CHF, evidence for disease modifying treatment is lacking. Clinical trials currently in progress to determine the effectiveness of ACE inhibitors and angiotensin II receptor antagonists in the management of diastolic CHF may clarify the prognosis and management of this condition. PMID- 12528969 TI - 'The second greatest benefit to mankind'? AB - In 1739 Samuel Johnson wrote an essay on the life of Dr Hermann Boerhaave, Professor of Physic at the University of Leiden, who died in 1738. Boerhaave, born 11 years after Harvey's death, could be said to have been influenced by Harvey in that he favoured experimental natural philosophy as the gateway to scientific medicine. He was denied entry into the church because he was accused wrongly of being a follower of the philosopher Baruch Spinoza, regarded as a heretic because he criticised established religious practices; this in spite of strongly supporting the love of God and humanity. Boerhaave decided to become a physician as he was, in Johnson's words, 'equally qualified for a profession, not indeed of equal dignity or importance, but which must undoubtedly claim the second place amongst those which are the greatest benefit to mankind'. It is this claim that I wish to examine. Can we still claim this regard for our profession? Is the medicine we practise, and the way we practise, of the greatest benefit to mankind, and how do we ensure that it is? PMID- 12528970 TI - New insights into hepatitis C. AB - Hepatitis C infection is characterised by three key features, which are the consequence of a complex interaction between genetic determinants of immune and other host factors and viral characteristics: 1. A high rate of viral persistence after acute infection resulting from a combination of weak T cell responsiveness and specific viral mechanisms of immune escape. 2. Marked interindividual variability in end-organ damage (fibrosis and cirrhosis), probably due to host genetic polymorphisms in genes governing the immune response and fibrosis pathways in addition to viral pathogenicity factors. 3. Significant resistance to antiviral therapies. Viral mechanisms of antiviral resistance parallel those of viral persistence, and include the intriguing possibility that hepatitis C may infect immunologically privileged sites such as the central nervous system. PMID- 12528971 TI - The impact of future technology on cancer care. AB - Globally cancer will increase greatly over the next 20 years because of ageing populations. Minimally invasive surgery will reduce the need for routine organ resection. The application of sophisticated computer systems to radiotherapy planning will allow the precise shaping of beam delivery conforming exactly to the shape of the tumour. The most promising advances will come from the rapidly increasing understanding of the molecular genetics of cancer. This will have considerable impact on prevention, screening, diagnosis and treatment and lead to a golden age of drug discovery. Individual cancer risk assessment will provide messages tailored for individual prevention and have far-reaching public health consequences. Increased consumerism in medicine will produce increasingly informed and assertive patients seeking out novel therapies, bypassing traditional referral pathways through global information networks. This will bring new ethical and moral dilemmas. The cancer future will be created by the interaction of four complex factors: technological success, society's willingness to pay, future healthcare delivery systems and the financial mechanisms that underpin them. PMID- 12528972 TI - Treatment in art. PMID- 12528973 TI - Finding a visual language for pain. PMID- 12528974 TI - Believing is seeing. PMID- 12528975 TI - Understanding pain: the enigma of pain and suffering. PMID- 12528976 TI - Neurology for the twenty-first century. PMID- 12528977 TI - William Harvey and his legacy. PMID- 12528978 TI - Harvey, clinical medicine and the College of Physicians. AB - This article deals with the problems of seeing Harvey historically, rather than as a construction seen from the viewpoint of modern medicine. It deals with his programme of work, the expectations of his audience, his intellectual training and the political and religious circumstances of seventeenth century Europe. It shows that at the time the impact of Harvey's discovery was negative on clinical medicine and its theory, but also shows ways in which that impact was favourable. PMID- 12528979 TI - Transparency, deception and trust. PMID- 12528980 TI - Managing medical emergency admissions. PMID- 12528981 TI - Shakespeare the physician. PMID- 12528982 TI - Ethics approvals and quagmires. PMID- 12528983 TI - Screening for thyroid disease in pregnancy: an audit. PMID- 12528984 TI - One stop swallowing clinic may be more effective than a new cancer clinic. PMID- 12528985 TI - Molecular aspects of drug recognition by specific T cells. AB - Adverse reactions to drugs are a major problem in pharmacotherapy. About 1/6 of all side-effects are thought to be drug-induced immune-mediated reactions. It is well established that T and B cells recognize a drug if it is bound as hapten to carrier molecules. However, the model does not explain many T cell-mediated reactions with chemically inert compounds. This review will first discuss the hapten-carrier concept of drug-presentation to T cells and the currently used methods to predict an allergenic potential of a drug. It then introduces our new model of drug-uptake- and processing-free HLA class II-restricted T-cell response termed "direct metabolism-independent T-cell stimulation". This led us to an other new concept: the pharmacological interaction of drugs with immunological receptors, namely the MHC and T-cell receptors. Additionally, we focus on certain conditions of non-covalent drug presentation by antigen presenting cells and on the molecular recognition of MHC/peptide/drug complexes by specific T-cell receptors. Finally, we discuss the clinical relevance of drug-specific T cells, namely that T cells seem to exert a certain pathology (e.g. drug-induced exanthema or pustular eruptions) depending on their function. These findings, which are based on the analysis of clinical drug allergy, have major implications for our understanding of T-cell biology and on the concept how to test and predict the allergenic potential of a drug. PMID- 12528986 TI - Emerging drug toxicities of highly active antiretroviral therapy for human immunodeficiency virus (HIV) infection. AB - To provide an overview of the epidemiologic parameters of emerging adverse effects associated with antiretroviral therapy for human immunodeficiency virus (HIV) disease. All available antiretroviral agents are associated with significant adverse drug effects. Of particular interest are newly emerging suspected adverse drug effects which were not generally noted in pre-marketing trials nor captured under current standard clinical care practices. Suspected antiretroviral toxicities meeting these criteria include: HIV-associated lipodystrophy which can include peripheral lipoatrophy, lipohypertrophy and metabolic abnormalities; hyperlactatemia and lactic acidosis; and metabolic bone abnormalities such as decreased bone mineral density, osteoporosis and osteonecrosis. Results of prospective and observational studies reported to date suggest that these abnormalities, while aetiologically complex, are likely attributable to treatment factors and may be intricately interrelated. The medical management of these symptoms remains unsatisfactory given the unexplored efficacy of traditional approaches in the HIV positive population. While the pathogenic mechanism of these disorders remains obscure, a theory of tissue specific mitochondrial toxicity has been proposed. With the continued introduction of novel therapies and standard treatment with combination therapy, new adverse events will continue to emerge among persons being treated for HIV disease. Beyond their immediate clinical implications, these events may contribute to changing patterns of antiretroviral utilisation including therapy initiation, adherence and cessation. PMID- 12528987 TI - Lipids as a target for drugs modulating multidrug resistance of cancer cells. AB - In this review we focus on the role of the membrane lipids in multidrug resistance and its modulation. Results of the research performed in recent years indicate the importance of lipid phase playing active role in many membrane processes. Along with the alterations of lipid membrane composition of cancer cells (with respect to the normal ones) the resulting changes of the biophysical membrane properties are discussed. Next we describe the general features of multidrug resistance phenomenon paying a special attention to the role of lipids and alterations of lipid membrane composition in MDR cells. Taking into account the phase separation properties of sphingolipids the importance of membrane heterogeneity (presence of caveole and lipid rafts) is emphasised. On the basis of vacuum cleaner hypothesis of drug transport proteins action we discuss the importance of lipid bilayer as medium in which diffusion of drugs takes place. Considering the membrane fluidity and its influence on the integral proteins activity, we underline the role of balance between the passive cellular influx and active efflux of the drug molecules. Finally the effects exerted on membranes by different kinds of multidrug resistance modulators (chemosensitizers) are described. In this part we discuss the influence of verapamil, phenothiazine derivatives, tamoxifen and chosen flavonoids on the biophysical properties of membrane lipids. Some further consequences of the alteration of membrane state are also considered. PMID- 12528988 TI - New multidrug resistance reversal agents. AB - Multidrug resistance (MDR) to antitumor agents represents a significant challenge to effective chemotherapy. The use of MDR modulators is a promising approach to overcome the undesired MDR phenotype. The more effective MDR modulators are urgently needed for clinical use. This review focuses on literatures published in 1998-2001. PMID- 12528989 TI - Cancer chemoprevention drug targets. AB - Cancer chemoprevention is a new approach in the management of cancer. Traditional cytotoxic chemotherapeutic approaches cannot cure most advanced solid malignancies. Chemoprevention can be defined as the use of non-cytotoxic drugs and natural agents to block the progression to invasive cancer. Chemoprevention can either prevent DNA damage that initiates the neoplastic transformation process or reverses the progression of pre-invasive lesions. Epidemiological observations, experimental evidence from animal carcinogenesis models, knock-out models, cancer cell lines and clinical trials have shown the efficacy of this approach. Recent advances in our understanding of carcinogenesis have led to the synthesis of new drugs that target specific receptors. Non-steroidal anti inflammatory drugs target the prostaglandin pathway. The identification of the role of cyclooxygenase-2 in epithelial carcinogenesis led to the synthesis of selective cyclooxygenase-2 inhibitors (Celecoxib). Celecoxib was subsequently approved for the prevention of colon polyps in familial adenomatous polyposis after the completion of a randomized clinical trial. The large chemoprevention clinical trial with the selective estrogen receptor modulator, tamoxifen, showed the benefit of tamoxifen in the prevention of breast cancer in high-risk women. Retinoids and rexinoids target the retinoid receptors and have a role in chemoprevention of aerodigestive, hepatic and cervical neoplasia. Selenium, an inhibitor of the glutathione peroxidase system, is being tested in the chemoprevention of prostate cancer and lung cancer. The different isoforms of vitamin E (tocopherols) may be chemopreventive. Recent evidence indicates that gamma-tocopherol may be a more powerful chemopreventive than the alpha tocopherol. The review details the rationale, experimental and clinical evidence and the drug targets of the chemopreventive agents that are currently in various phases of clinical development. PMID- 12528990 TI - Central oxytocinergic neurotransmission: a drug target for the therapy of psychogenic erectile dysfunction. AB - A group of oxytocinergic neurons originating in the paraventricular nucleus of the hypothalamus and projecting to extrahypothalamic brain areas (e.g. hippocampus, medulla oblongata and spinal cord) control penile erection. Activation of these neurons by dopamine and dopamine agonists, excitatory amino acids (N-methyl-D-aspartic acid) or oxytocin itself, or by electrical stimulation leads to penile erection, while their inhibition by GABA and GABA agonists or by opioid peptides and opiate-like drugs inhibits this sexual response. The activation of oxytocinergic neurons in the paraventricular nucleus by dopamine, oxytocin and excitatory amino acids is apparently secondary to the activation of nitric oxide (NO) synthase. NO in turn activates, by a mechanism that is as yet unidentified, the release of oxytocin from oxytocinergic neurons in extrahypothalamic brain areas. Several peptide analogues of hexarelin, a growth hormone releasing peptide, also induce penile erection when injected into the paraventricular nucleus and, to a lesser extent, systemically, apparently by acting on a specific receptor to activate oxytocinergic neurons as shown for the above drugs and oxytocin. Paraventricular oxytocinergic neurons and mechanisms similar to those reported above are also involved in the expression of penile erection in physiological contexts, namely when penile erection is induced in the male by the presence of an inaccessible receptive female, which is considered a model for psychogenic impotence in man, as well as during copulation. These findings show that paraventricular oxytocinergic neurons projecting to extra hypothalamic brain areas and to the spinal cord are a likely target for the treatment of erectile dysfunction of central origin. PMID- 12528991 TI - Membrane receptors for vitamin D steroid hormones: potential new drug targets. AB - There is increasing evidence that steroid hormones derived from vitamin D act through classical nuclear receptors (nVDR), as well as specific binding sites on the plasma membrane of target cells that are coupled to signal transduction systems. These sites are referred to as Membrane Associated, Rapid Response Steroid (MARRS) binding proteins or complexes. In the case of the seco-steroid 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], the two 'receptors' appear to be different proteins with distinct affinities for vitamin D analogs. These differences may be useful in a number of clinical situations. In treating leukemias, it would be useful to promote the actions of the nVDR for differentiation to macrophages while blocking the 1,25D3-MARRS in intestine, which might contribute to the undesirable side effect of hypercalcemia. In contrast, stimulation of the intestinal 1,25D3-MARRS would be desirable in the elderly, since this signalling system appears to decline with age in model systems, potentially contributing to diminished intestinal absorption of calcium and associated bone loss. Bone itself is known to have osteoblasts that respond to 1,25(OH)2D3 through both nVDR and 1,25D3-MARRS mechanisms. Both systems are required for bone-building activities. Osteoclasts lack the nVDR, but may become activated through the 1,25D3-MARRS, offering another site of drug intervention in the treatment of osteoporosis. Finally, during tooth mineralization, immunohistochemical studies reveal an absence of the nVDR and a marked appearance of the 1,25D3-MARRS. In addition to our growing knowledge of 1,25(OH)2D3, the physiological actions of a lesser studied metabolite of vitamin D, 24,25(OH)2D3, are coming to light and may offer additional targets for pharmaceutical modulation. PMID- 12528992 TI - Phytoestrogens: pharmacological and therapeutic perspectives. AB - Phytoestrogens exert different estrogen receptor-dependent and -independent pharmacological actions. They share with estrogens several structural features and show greater affinity for the newly described estrogen receptor-beta. Many hope that phytoestrogens can exert the cardioprotective, anti-osteoporotic and other beneficial effects of the estrogens used in hormone replacement therapy in postmenopausal women without adversely affecting the risk of thrombosis and the incidence of breast and uterine cancers. Although there are many positive indications that phytoestrogens can fulfil this role, it remains to be proven: controlled interventional studies are lacking, and many questions remain unanswered. This review analyzes, on the basis of available experimental and epidemiological studies, the pros and cons of phytoestrogen use and describes the potential tissue targets and mechanisms of action of phytoestrogens. PMID- 12528993 TI - The Twenty-Third Sir Harold Gillies Memorial Lecture. Keeping a head. PMID- 12528994 TI - Paediatric transcranial surgery: a review of 114 consecutive procedures. AB - Craniofacial units have a responsibility to collect data, to promote research and training and to carry out audit. We present a review of 114 consecutive transcranial procedures performed in 110 children in our unit over an 8 year period, with particular reference to complications. There were no deaths in this series. Complications included two cases of excessive intraoperative blood loss necessitating a delay in the procedure, and two postoperative infections that required aggressive antibiotic management. Minor complications, delaying hospital discharge, occurred in 13 patients. Within the range of paediatric transcranial procedures performed, the potential for complications is greater for complex osteotomies in syndromic conditions than for single sutural synostosis correction. The transcranial case mix included a relatively small number of craniofacial dysostoses, which contributes to the very low complication rate reported. This report demonstrates that multidisciplinary assessment and planning, adherence to craniofacial surgical principles, shorter operating times and avoidance of high-risk procedures contribute to a low complication rate, and confirms that paediatric transcranial procedures can be safely performed in dedicated centres where there is a multidisciplinary team with appropriate commitment and experience. PMID- 12528995 TI - Crab-pincers style facial artery flap. AB - To reconstruct defects as large as 5 cm in diameter in the region extending from the columella and anterior nasal floor to the upper lip, we use a crab-pincers style facial artery (CPFA) flap technique combining nasolabial flaps and cheek advancement flaps. In the CPFA flap, the bifurcation of the facial artery allows the angle between the nasolabial flap and the cheek advancement flap to be freely altered in the manner of crab pincers. By combining the four leaves of bilateral CPFA flaps at the centre, appropriate reconstruction of the three-dimensional structures surrounding the columella can be achieved. In addition, this method requires only one operation to complete extensive reconstruction. The method does not result in adverse scarring or scar contracture. After the procedure,appropriate moustache growth gives a natural impression and conceals philtral distortion. We consider the CPFA flap to be very useful for reconstructing large defects in the central facial region. PMID- 12528996 TI - Head and neck oncology: the UK experience. Who is publishing what? AB - Using the MEDLINE database (OVID), a retrospective review of the UK literature on head and neck oncology was performed for the period 1994-2000, each publication being categorised by department and first author. Tumours almost exclusively managed by general surgeons, cardiothoracic surgeons and neurosurgeons were excluded. In the years 1994 to 2000, there were a total of 120 UK publications, 72% of which came from non-academic NHS units; 23% of the publications were from ENT units, 23% from oral and maxillofacial (OMF) surgery units and 18% from plastic surgery units. The majority of plastic surgery publications described reconstructive techniques, whereas a wider range of topics was observed in the publications by ENT and OMP surgeons. Several irregularities in the MEDLINE database were discovered and are discussed. The findings of this study may be relevant to the future planning of head and neck oncology services. PMID- 12528998 TI - The combined posterior temporoparietal and galeal fascial flap: a new flap in the elevation of the constructed auricle (second stage of microtia correction). AB - Over the past few years, the technique of elevating the buried ear framework in the second stage of microtia correction has shifted from skin grafting to the use of flaps and cartilage blocks in the retroauricular sulcus. While the temporoparietal fascial flap should be reserved for secondary procedures and the treatment of complications, the mastoid fascial flap is inadequate by itself and needs an additional cartilage graft. Here, we describe a new flap, the combined posterior temporoparietal and galeal fascial flap, for the elevation of the buried ear cartilage. The flap is robust, with a dependable blood supply based on the posterior branches of the superficial temporal artery. In four cases the flap was rolled up and inset into the retroauricular sulcus, while in three cases an additional conchal cartilage graft was inserted into the roll. All the patients had satisfactory ear projection at follow-up 10-14 months postoperatively. We discuss the surgical technique and the advantages of this flap. We believe that this new flap, which has not been described before, has the potential to replace other flaps in the second stage of microtia correction. PMID- 12528997 TI - Fading of nipple-areolar reconstructions: the last hurdle in breast reconstruction? AB - Fading of nipple-areolar reconstructions is commonly reported, but there are few formal studies of this phenomenon. The purpose of this study was to determine whether deficiencies in nipple-areolar reconstruction and pigmentation were perceived by patients, their partners and independent observers, and whether a technique could be developed to measure nipple-areolar colour reliably. A total of 57 patients, 32 partners and four independent observers completed questionnaires about the appearance of the patients' breast reconstructions in general and specifically about their nipple-areolar reconstructions. Scores for the general attributes of the breast reconstruction were used as internal controls for the scores of the nipple-areolar reconstruction. A computer software package was developed to analyse colour in photographs of the reconstructions. Independent observers thought that nipple-areolar reconstruction improved the appearance of a breast reconstruction 81% of the time. Considerably fewer patients were happy with their nipple-areolar colour than were happy with the more general attributes of the breast reconstruction (P < 0.005). Colour analysis objectively demonstrated measurable mismatch between normal and reconstructed nipple-areolar skin, which was positively correlated with time since surgery due to fading of the nipple-areolar reconstruction. In our patients, the quality of nipple-areolar reconstruction, in particular its pigmentation, is seen as inferior to that of the rest of the breast reconstruction in the eyes of patients, their partners and independent observers. The poor colour match and fading of reconstructed nipple-areolar skin are phenomena that can be measured using colour analysis. PMID- 12528999 TI - Salvage of major joint prostheses in aggressive non-specific panniculitis. AB - Panniculitis is a relatively rare condition, which is usually treated non operatively. We present a case of panniculitis causing an acute septicaemic episode and threatening the integrity of underlying joint prostheses. The situation was salvaged by radical debridement of the affected areas, and reconstruction using split-skin grafts. Considerations in the management of patients with this condition are discussed. PMID- 12529000 TI - The recovery suture. PMID- 12529001 TI - Titanium clips: a simple technique for the excision of accessory tragi and digits. PMID- 12529002 TI - Ear reconstruction in elderly patients: a two-part helix method in a framework. PMID- 12529003 TI - A maternity bra as a dressing after breast reduction. PMID- 12529004 TI - The impact of breast augmentation on the breast-screening programme. PMID- 12529005 TI - An intraoperative DIY breast sizer. PMID- 12529006 TI - The use of image-editing software in plastic surgery teaching. PMID- 12529007 TI - The stretch test in basal cell carcinoma: a clinical indicator of tumour. PMID- 12529008 TI - If the shoe fits. PMID- 12529009 TI - Behaviour of cancellous bone graft placed in induced membranes. PMID- 12529010 TI - Gillies' forceps diathermy burns: a cautionary note. PMID- 12529012 TI - Candida glabrata, an emerging fungal pathogen, exhibits superior relative cell surface hydrophobicity and adhesion to denture acrylic surfaces compared with Candida albicans. AB - Oral candidosis is a common opportunistic infection in debilitated individuals and Candida glabrata is the second most frequently isolated species from this condition, after Candida albicans. Candidal adherence to various biological or non-biological surfaces is considered a prerequisite for colonization, and pathogenesis of candidal infections, and their relative cell surface hydrophobicity (CSH) is likely to be a possible contributory force involved in this process. Whereas a large body of data on the latter features of C. albicans is available, there is surprisingly little information on C. glabrata. As a comprehensive database on the relative adhesion and CSH of Candida spp. is instructive and useful, we investigated in vitro the latter attributes of 34 oral isolates of C. glabrata and 15 isolates of C albicans. There were remarkable intraspecies differences in both the CSH and the adhesive ability of C. glabrata strains (p < 0.001). Compared with C. albicans, C glabrata demonstrated a four fold greater CSH value (30.63 +/- 11.20% vs 7.23+/-3.56%, p < 0.0001) and a two fold greater tendency to adhere to denture acrylic surfaces (75.18 +/- 39.96 vs 30.36+/-9.21, p < 0.0001). A significant positive correlation between CSH and adhesion was also noted for both C. glabrata (r=0.674, p < 0.0001) and C. albicans ( r = 0.636, p < 0.05). When the effect of different incubation conditions on the relative CSH and adherence of C. glabrata was examined, CSH and the adherence to acrylic surfaces of four of six C. glabrata isolates were significantly affected by a reduction of the culture temperature (from 37 degrees C to 25 degrees C). A positive relationship also emerged when the temperature induced variations in the adherence values were correlated with their relative CSH. These data provide hitherto unavailable archival information on important pathogenic attributes of the two most common oral Candida species that may help explain their predominance in this milieu. PMID- 12529011 TI - Difference in neutrophil cytokine production induced by pathogenic and non pathogenic mycobacteria. AB - This study shows differences between Mycobacterium tuberculosis and Mycobacterium avium (opportunistic) and Mycobacterium smegmatis (non-pathogenic), with respect to their abilities to induce cytokine/chemokine release from human neutrophils. Neutrophils incubated with live cells of M. tuberculosis, M. avium, or M. smegmatis produced and released TNF-alpha, IL-6, and IL-8. No or very small amounts of these cytokines/chemokines were found in resting neutrophils, suggesting that they were newly synthesised. The levels of TNF-alpha, IL-6, and IL-8 produced/released from neutrophils incubated with M. tuberculosis were markedly lower than those of the opportunistic or non-pathogenic bacterial species. The production of TNF-alpha reached a maximum level at a time (4 h) when the production of IL-8 had only just started, and this was true for all three mycobacteria tested. However, the time course for IL-6 production differed between the species, reaching a peak value after 8 h for M. tuberculosis not seen with the other bacteria. It is likely that the relatively high levels of cytokines induced by opportunistic/non-pathogenic mycobacteria are of importance for the induction of an innate immune response through which these organisms are eliminated, while the low levels of cytokines released by neutrophils interfering with M. tuberculosis might help the bacteria to persist. PMID- 12529013 TI - GLUT4 expression at the plasma membrane is related to fibre volume in human skeletal muscle fibres. AB - In this study we examined the relationship between GLUT4 expression at the plasma membrane and muscle fibre size in fibre-typed human muscle fibres by immunocytochemistry and morphometry in order to gain further insight into the regulation of GLUT4 expression. At the site of the plasma membrane, GLUT4 was more abundantly expressed in slow as compared to fast fibres at the same fibre diameter (p < 0.01) and the GLUT4 expression increased with increasing fibre radius independently of fibre type (p < 0.01). The GLUT4 density at the surface of slow fibres of both diabetic and obese was reduced compared to control subjects at the same diameter (p < 0.001). Fast fibres in obese and type 2 diabetic subjects expressed a fibre-volume-dependent GLUT4 expression (p < 0.001), while this did not reach significance in slow fibres (obese p = 0.18 and diabetic p = 0.06). Our results show that increasing fibre volume is associated with increasing GLUT4 expression in both slow and fast fibres. Based on the possible dependency of GLUT4 expression on volume, we hypothesize that the reduced GLUT4 expression in obesity and type 2 diabetes may partly be compensated for by physical activity. PMID- 12529014 TI - Hepatitis B virus infection in drug addicts: no acute fatalities, no chronicity and could have benefits. AB - In a group of 159 drug addicts with acute hepatitis B in the 1970s there were no fatalities. During an observation period of about 25 years, 51 of the 159 died of various causes, but no deaths could be attributed to the hepatitis B infection. From 1998 to 2001, a follow-up examination of 53 of the 108 patients still alive, none of the 53 had a chronic hepatitis B virus infection. Fifteen out of thirty five patients who had completely stopped using narcotics claimed that hospitalisation for acute hepatitis B had been an important factor in their decision to quit drugs. PMID- 12529015 TI - Bacillarity at autopsy in pulmonary tuberculosis. Mycobacterium tuberculosis is often disseminated. AB - The aim of this investigation was to quantify dissemination of Mycobacterium tuberculosis infection in patients with pulmonary tuberculosis and to show the pattern of eradication during treatment. The study is based on 98 out of the 113 patients with pulmonary tuberculosis who died during their admission to hospital in the Municipality of Copenhagen from 1963 to 1971. These patients had cultures for M. tuberculosis performed from different organs at autopsy: 78% treated <=100 days had dissemination of bacteria, cultured with decreasing frequency in the lungs, spleen, liver, and kidneys, respectively. In comparison, 23% treated >100 days had dissemination of bacteria, among which 50% occurred in patients with records of poor treatment compliance, 14% in patients with good treatment compliance. 81% of all patients had at least one chest x-ray judged to be without a miliary pattern. This study emphasizes that M. tuberculosis is often disseminated to organs other than the lungs in severe pulmonary tuberculosis. Eradication of bacteria in these organs can take several months. This observation adds to our understanding of the natural history of tuberculosis: M. tuberculosis is a resilient organism that can adapt to a wide variety of environmental conditions. PMID- 12529016 TI - Serum tetranectin is an independent prognostic marker in colorectal cancer and weakly correlated with plasma suPAR, plasma PAI-1 and serum CEA. AB - Soluble tetranectin (TN) was measured preoperatively in serum from 567 patients with primary colorectal cancer and levels were tested for association with prognosis. The prognostic significance of TN was also compared to that of plasminogen-activator inhibitor-1 (PAI-1), urokinase plasminogen activator (uPAR) and carcinoembryonic antigen (CEA). Significantly shorter survival was found for patients with TN levels below a cut-off point of 7.5 mg/l compared to patients with levels above, as illustrated by Kaplan-Meier curves. By Cox analyses, log TN, log soluble uPAR as well as log CEA were found to have an independent prognostic value for survival (log TN: HR = 0.47, 95% CI: 0.29-0.76); log soluble uPAR: HR = 1.65, 95% CI: 1.18-2.31; log CEA: HR = 1.I1, 95% CI: 1.03-1.20). Based on the multivariate model, a patient with a combination of low levels of TN and PAI-1 and elevated levels of soluble uPAR and CEA had a 2.43 increased risk as compared to a patient with median levels of these biochemical markers. Significant correlations were found with Dukes' stages for all the biochemical markers and between the respective biochemical markers. The findings confirm that TN is a strong prognostic factor in patients with colorectal cancer. TN may be valuable as a prognostic variable in future studies evaluating new treatment strategies for colorectal cancer. PMID- 12529017 TI - Esthesioneuroblastoma: a Danish clinicopathological study of 40 consecutive cases. AB - Aims. To review all cases of esthesioneuroblastoma in Denmark from 1978 to 2000 with respect to staging, grading, histopathological and immunohistochemical evaluation, and prognostication. Methods and results. Possible cases of esthesioneuroblastoma were retrieved from Danish oncology departments. Patients were included on the basis of review of their files or pathology reports, and/or on the basis of histopathological and immunohistochemical examination. Forty-nine possible cases were retrieved. Nine cases were excluded. Esthesioneuroblastoma is a malignant neuroendocrine tumour originating in the olfactory mucosa. It is a small blue cell neoplasm with a characteristic lobular architecture. It has a neuroendocrine immunophenotype and a sustentacular S-100 staining pattern. The tumours were staged according to Kadish and graded according to Hyams. Kaplan Meier survival analysis was used to identify prognostic factors. Conclusion. The Kadish staging system was able to group the patients into prognostically relevant groups. Intracranial involvement and metastases at the time of diagnosis were found to be poor prognostic factors. Hyams grading system is difficult to work with and it was not possible to divide patients into prognostically relevant groups. Presence of necrosis, a diffuse growth pattern and a high proliferation index proved to be equally poor prognostic factors. PMID- 12529019 TI - Mutations in gyrA, gyrB, parC, and parE in quinolone-resistant strains of Neisseria gonorrhoeae. AB - Mutations in the genes for the subunits GyrA and ParC of the target enzymes DNA gyrase and topoisomerase IV are important mechanisms of resistance in quinolone resistant bacteria, including Neisseria gonorrhoeae. The target enzymes also consist of the subunits GyrB and ParE, respectively, though their role in quinolone-resistance has not been fully investigated. We sequenced the quinolone resistance-determining regions (QRDR) of gyrA, gyrB, parC, and parE in 25 ciprofloxacin-resistant strains from Bangladesh (MIC 4-->32 mg/l) and 5 susceptible strains of N. gonorrhoeae. All the resistant strains had three or four mutations. Two of these were at positions 91 and 95 of gyrA. Fourteen strains had an additional mutation in parC at position 91, and 17 strains had an additional mutation in parE in position 439. No alterations were found in gyrB. The five susceptible strains had identical DNA sequences. Data indicate that the mutations detected in the QRDR of gyrA and parC may be important in the development of quinolone resistance. According to transformation experiments we assume that the alteration in parE is not related to a high degree of quinolone resistance. There was no correlation between ciprofloxacin MICs and pattern or number of mutations in the target genes. PMID- 12529018 TI - VEGF concentrations in tumour arteries and veins from patients with rectal cancer. AB - This pilot study investigated the hypothesis that the tumour itself is the source of the elevated vascular endothelial growth factor (VEGF) concentrations which are often observed in peripheral blood from patients with rectal cancer. Twenty four consecutive patients with primary rectal cancer were included. Blood samples were drawn preoperatively from peripheral veins (I) and intraoperatively from peripheral veins (II), tumour arteries (III), and tumour veins (IV). In the four compartments, VEGF concentrations were measured in serum, EDTA plasma, and supernatants from lysed whole blood. Additionally, automated complete white cell and platelet counts were performed. In serum and EDTA plasma, no significant differences in VEGF concentrations were observed (p = 0.1 and p = 0.5), respectively) between tumour arteries and tumour veins. However, in supernatants from lysed blood, VEGF concentrations were significantly (p = 0.03) lower in venous blood than in arterial blood. Unexpectedly, a 16% reduction (p < 0.0001) in the number of neutrophils was observed during transit of the arterial blood through the rectal tumours, while none of the other types of leukocytes or platelets was significantly reduced in numbers during the same passage. These findings indicate that the tumour itself is not the only source of elevated VEGF concentrations in peripheral blood from patients with rectal cancer. A consistent finding was that a large number of neutrophils disappeared from the blood during passage through the rectal tumour. However, the significance and fate of the migrating neutrophils cells are unknown and should be investigated further. PMID- 12529020 TI - Neuroendocrine differentiation in carcinoma of the breast. Tyramide signal amplification discloses chromogranin A-positive tumour cells in more breast tumours than previously realized. AB - The aim of the study was to determine if, by means of tyramide signal amplification (TSA), the presence of chromogranin A (CgA)-positive tumour cells could be demonstrated in breast cancer cases found to be negative by conventional immunohistochemical staining. Sections from 44 cases of breast cancer (28 infiltrating ductal carcinomas, 2 lobular carcinomas, 4 ductal carcinomas in situ (DCIS), 7 lobular carcinomas in situ (LCIS), and 3 mucinous carcinomas) were stained for CgA by conventional immunohistochemical methods and by immunohistochemistry with TSA. The sections were also histologically graded and their oestrogen receptor (ER), progesterone receptor (PgR) and HER-2 oncogene status was recorded. Five of the tumours showed CgA-positive staining with the polyclonal antibody 430 with conventional methods. Thirty cases showed CgA immunoreactive tumour cells after immunohistochemical staining with the polyclonal antibody 430 with TSA. However, eight of these also showed faint staining with the negative control antibody X0936 with TSA. One case showed immunopositivity for CgA using a monoclonal antibody without tyramide amplification and only a further two cases were positive when TSA was applied. The presence of CgA appears to be associated with a lower histological grade and may be more often found in oestrogen receptor-positive tumours. PMID- 12529021 TI - Clonal clustering and colonization factors among thermolabile and porcine thermostable enterotoxin-producing Escherichia coli. AB - A considerable proportion of enterotoxigenic Escherichia coli (ETEC) do not possess identifiable colonization factors (CFs). Genetic fingerprint analyses based on repetitive sequence-based polymerase chain reaction (rep-PCR) showed that 9 of 10 such CF-negative isolates which produced the thermolabile and the porcine thermostabile enterotoxin could be divided into three clusters. Following transformation with a plasmid harbouring the gene encoding CfaR, a positive regulator for several ETEC adhesins, three of the six strains in the first cluster expressed coli surface antigen 20 (CS20). No CFs were identified on the two transformed strains in the second cluster while the transformants of the two strains in the last cluster expressed CS12, the N-terminal amino acid sequence of which was deciphered. The study illustrates the potential of using genetic fingerprinting to group ETEC into clusters of strains with genes encoding different CFs and confirms the ability of CfaR to induce the expression of several different CFs. PMID- 12529022 TI - The future of image perception in radiology: synergy between humans and computers. PMID- 12529023 TI - Perception of breast cancer: eye-position analysis of mammogram interpretation. AB - RATIONALE AND OBJECTIVE: The author performed this study to determine how image based elements are translated into decisions by radiologists with different levels of experience in the reading of mammograms. MATERIALS AND METHODS: Three full-time mammographers and four radiology residents read 40 two-view mammogram cases. The observers' eye position was tracked while they searched the mammograms for malignancies. Spatial frequency analysis was performed to relate what the observers reported with where they looked. RESULTS: Statistically significant differences were found between lesion-containing areas that attracted visual attention and were correctly interpreted and those that were visually inspected but not reported. In addition, an artificial neural network was successfully trained to map the image characteristics in the visually selected areas on a mammogram and to linkthem to a likely decision by the observer. CONCLUSION: Spatial frequency analysis can be used to derive trends for how mammographers and radiology residents will respond to mammograms. PMID- 12529025 TI - Validation of the summary ROC for diagnostic test meta-analysis: a Monte Carlo simulation. AB - RATIONALE AND OBJECTIVES: The author performed this study to test a technique for validating the logit regression method for summary receiver operating characteristic (ROC) meta-analysis, perform initial validation studies, and identify areas for further investigation. MATERIALS AND METHODS: Monte Carlo simulation was performed by using a custom macro program for a personal computer spreadsheet. The program creates simulated data sets based on user-specified parameters, performs a meta-analysis on the data sets, and logs the results so the accuracy and variability of the method can be measured. The program can also be used to measure the effects of changes in study design and meta-analysis parameters. RESULTS: For the base case of a small meta-analysis (10 studies) of small trials (mean, 50 patients), the meta-analysis results closely matched the input sensitivity and specificity when they were less than 80%. Systematic errors, if any, were small. At sensitivities and specificities greater than 80%, the true sensitivity or specificity was underestimated by up to 2% in the meta analysis. Confidence intervals calculated with the summary ROC curve were reasonably conservative, although they too fell below the true results when sensitivity or specificity was greater than 80%. The underestimation was eliminated when the simulation was repeated for a much larger trial (mean, 1,000 cases per study)--even with a sensitivity and specificity of 98%. CONCLUSION: The Littenberg-Moses method for summary ROC meta-analysis is effective for obtaining an accurate summary estimate of diagnostic test performance, although the continuity correction introduces a small downward bias in the meta-analysis of small trials. PMID- 12529024 TI - System for upper airway segmentation and measurement with MR imaging and fuzzy connectedness. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to evaluate whether a computerized system developed to help delineate the upper airway and surrounding structures with magnetic resonance (MR) imaging was effective for aiding in the diagnosis of upper airway disorders in children. MATERIALS AND METHODS: The authors performed axial T2-weighted MR imaging to gather information about different aspects of the airway and its surrounding soft-tissue structures, including the adenoid and palatine tonsils, tongue, and soft palate. Images were processed and segmented to compute the architectural parameters of the airway (eg, surface description, volume, central [medial] line, and cross-sectional areas at planes perpendicular to the central line). The authors built a software package for the visualization, segmentation, registration, prefiltering, interpolation, standardization, and quantitative analysis of the airway and tonsils. RESULTS: The system was tested with 40 patient studies. For every study, the system segmented and displayed a smooth three-dimensional rendition of the airway and its central line and a plot of the cross-sectional area of the airway orthogonal to the central line as a function of the distance from one end of the central line. The precision and accuracy for segmentation was 97%. The mean time taken per study was about 4 minutes and included the operator interaction time and processing time. CONCLUSION: This method provides a robust and fast means of assessing the airway size, shape, and level of restriction, as well as a structural data set suitable for use in modeling studies of airflow and mechanics. PMID- 12529026 TI - PACS utilization in radiologic research. AB - RATIONALE AND OBJECTIVES: The authors investigated patterns of utilization of the digital picture archiving and communication system (PACS) in radiologic publications in order to ascertain whether there may be a correlation between PACS use and demographic/cultural factors such as geographic location, radiologic specialization, and use of advanced radiologic technologies. MATERIALS AND METHODS: A total of 1,037 articles in 22 randomly selected issues of AJR: American Journal of Roentgenology and of Radiology (1999-2001) were reviewed for the rate of PACS use and the type of PACS used. Articles for which PACS use or nonuse was established were further classified according to originating continent or region, imaging modality, study design (whether retrospective or prospective), and the use of advanced radiologic technology. The use of a PACS was then correlated with these factors. The data were recorded and statistics were prepared by means of statistical software. The nonparametric (chi2) test also was run by using this software. RESULTS: PACS had been used and reported in the preparation of 225 of the 1,037 articles. The type of PACS used was mini PACS (eg, systems using digital imaging and communications in medicine [DICOM] protocols or precursors) in 55 (24%) and department- or hospital-wide PACS in 161 (72%). Most of the articles for which use of a PACS was reported had originated in North America (60%), Europe (22%), or Asia (14%). PACS were used in almost half of retrospective studies and in one-fourth of prospective studies (P < .01). A low correlation was found between utilization of PACS and use of other advanced technologies. CONCLUSION: Although PACS were utilized in the preparation of a substantial proportion of articles published in the two major radiology journals, there was a great disparity in the rate of PACS use among world regions. The proportion of studies originating in North America for which a PACS was used was nearly three times the number of similar studies originating in Europe, and more than four times the number originating in Asia. PMID- 12529027 TI - Animal-based model to investigate the minimum tumor size detectable with an electrical impedance scanning technique. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to determine the minimum tumor size detectable with electrical impedance scanning (EIS) in laboratory animals. MATERIALS AND METHODS: VX2 tumor cells (1 mm3) were implanted bilaterally into the upper leg musculature of five white New Zealand rabbits. EIS and ultrasound (US) were performed before, during, and immediately after implantation and on every 2nd day thereafter until tumors could be visualized with both modalities. This was followed by an extended follow-up regimen until a tumor size of 1 cm3 was reached. Rabbits were anesthetized subcutaneously. RESULTS: Tumors could not be implanted in one rabbit. Neither EIS nor US were performed in one rabbit due to severe skin alterations at the implantation site. No focal increase in conductance was visible before or immediately after tumor injection. The smallest tumor identified with EIS was determined with US to be approximately 8 mm3. The mean tumor size at initial detection was 52 mm3 (determined with EIS). In two cases, the tumor was first seen with US (EIS detection was delayed with a delay of 1 and 2 days, respectively). In all remaining cases, tumors were detected simultaneously with both EIS and US. All tumors were visible with EIS. CONCLUSION: The animal-based model is feasible. VX2 tumors are detectable with EIS. Tumors characterized by a focal spot at EIS could be detected starting at a tumor size of 7.5 mm3. PMID- 12529028 TI - Factors that influence radiologists' career choices. AB - RATIONALE AND OBJECTIVES: The authors performed this study to examine the factors, particularly the modifiable factors, that influence the career choices of radiologists immediately after graduation from residency and later. MATERIALS AND METHODS: A survey was sent to 119 radiologists who had graduated from a large academic training program between 1981 and 2000. The graduates were asked to classify their first job and any subsequent jobs in academic radiology or private practice and to identify the reasons for their initial job choices and any job changes. A nested cohort study was performed to evaluate the effect of research experience on career choice. RESULTS: Seventy-nine (66%) graduates responded to the survey. Forty-three (54%) of the respondents had chosen academic positions as their first jobs. Those who had published during their residency were 26.4 times more likely to choose an academic position as a first job. Twenty-four graduates had since left their academic jobs for private practice. Although the discrepancy in financial rewards between academic radiology and private practice was the main reason for the job switch in 71% of these cases, 33% of the respondents cited difficulty with research as a reason. In addition, only 25% of current academic radiologists were satisfied with their research activities. CONCLUSION: An exodus from academic radiology to private practice is evident among graduates from this large academic residency program, with greater financial reward being the primary motivation. However, a positive research experience during residency could persuade more graduates to choose and to continue in an academic career. PMID- 12529029 TI - Dynamic quiz bank: a portable tool set for authoring and managing distributed, Web-based educational programs in radiology. AB - RATIONALE AND OBJECTIVES: The authors performed this study to evaluate a portable, platform-independent software program that enables users from remote sites to transform raw materials (eg, text, images, video) into Web-ready, interactive tutorials and examinations. MATERIALS AND METHODS: The software program evaluated consists of three modules: a network-based interface developed in the Java programming language, an Oracle 8i relational database with liaison software, and shell scripts developed in the Perl programming language to automate the authoring, maintenance, and updating of content in a dynamic quiz bank (DBQ). Four faculty members, one radiology resident, and two undergraduates majoring in computer science volunteered to create questions for the DQB and to evaluate ease of authoring. RESULTS: Faculty members with various levels of computer proficiency were able to establish a comprehensive DQB of more than 1,000 interactive questions. These radiologists reported the scripts reliable and easy to use. The DQB, offered in a pathology course for 2nd-year medical students, was used by 151 students and may have played a role in improving standardized test scores. Eighty-seven percent (n = 131) of the students believed that the DQB was extremely useful as an educational tool. CONCLUSION: The DQB software program facilitated access to, and authoring and maintenance of, Web based educational materials developed in the departments of radiology and pathology. PMID- 12529030 TI - Optimization of TI values in inversion-recovery MR sequences for the depiction of fine structures within gray and white matter: separation of globus pallidus interna and externa. AB - RATIONALE AND OBJECTIVES: The authors' purpose was to search the inversion time (TI) values that enable the best differentiation of fine structures in gray matter (gray-gray differentiation). MATERIALS AND METHODS: Seven healthy adult volunteers with no history of neurologic disease or head trauma were recruited and gave their informed consent. The subjects consisted of two men and five women ranging in age from 25 to 38 years, with a mean age of 28 years +/- 5. The subjects were imaged with a turbo spin-echo inversion-recovery sequence. This sequence was performed in the axial plane at the level of the basal ganglia with the following parameters: repetition time, 3,200 msec; echo time, 15 msec; three signals acquired; echo train, seven; section thickness, 3 mm; matrix size, 256 x 256; and field of view, 180 mm. The tested values were TI = 100, 200, 300, 400, and 500 msec. Region-of interest measurements were performed at the following anatomic structures and represent gray-gray and white-white differentiations, respectively: globus pallidus externa versus globus pallidus interna, and optic radiation versus surrounding white matter. RESULTS: The maximum contrast index value occurred at TI = 400 msec for globus pallidus externa versus globus pallidus interna (P < .05) With the contrast-to-noise ratio, no significant difference in gray-gray differentiation was observed among the various TIs. The minimum signal-to-noise ratio of the gray matter occurred at TI = 400 msec (P < .05). A subjective evaluation revealed an overall superiority of gray-matter differentiation with TI = 400 msec. CONCLUSION: A TI of 400 msec was the most suitable for this purpose. PMID- 12529031 TI - Radiologist's vision check no. 2. PMID- 12529032 TI - Parametric visualization methods for the quantitative assessment of myocardial motion. AB - RATIONALE AND OBJECTIVES: The authors performed this study to evaluate three dimensional (3D) and four-dimensional (4D) techniques for quantifying and visualizing myocardial motion. MATERIALS AND METHODS: The 4D method was performed by using 3D reconstructions of the complete, in vivo, canine heart before and after acute myocardial infarction. Images were obtained with the Dynamic Spatial Reconstructor (1-3) at 15 time points throughout one cardiac cycle. The authors used 0.75-mm-thick sections to allow creation of deformable models at each time point. For the 3D method, electron-beam computed tomographic reconstructions were obtained in anesthetized pigs from eight adjacent short-axis sections of the left ventricle. Data were acquired before and after selective microembolization of the left anterior descending coronary artery at 11 time points throughout one complete cardiaccycle. The authors used 8-mm-thick sections, which did not enable the use of the volumetric 4D approach with deformable models. For the 3D method, images were processed by radially dividing the tomographic images into small circumferential sectors. Color encoding was used for the derived local magnitudes of wall dynamics. RESULTS: The 4D method provided endocardial peak velocities, excursions, and strains throughout systole and diastole. The 3D method provided regional thickness or regional rates of left ventricular wall thickening throughout the cardiac cycle. CONCLUSION: Functional parametric maps of disturbances in regional contractility and relaxation facilitate appreciation of the effect of altered structure-to-function relationships in the myocardium. PMID- 12529033 TI - Medical student attitudes toward inclusion of a research year within diagnostic radiology residency: a survey of students participating in the 2002 NRMP match. National Resident Matching Program. AB - RATIONALE AND OBJECTIVES The purpose of this study was to solicit medical student opinion regarding establishment of a research track within diagnostic radiology residency and to determine the method of implementation most favorable to candidates who self-identified as future researchers. MATERIALS AND METHODS: After the deadline for submitting rank lists (February 21, 2002), a questionnaire was mailed to 588 U.S.-resident medical students who applied to the authors' residency in the 2002 match. Returned questionnaires post-marked before match day (March 21, 2002) were included in the analysis. Students were asked to rate their level of interest in different residency curricula; to indicate whether they were most likely to pursue a position in private practice, clinical academics, or research academics; and to indicate their preferred time of choosing a training track, preferred duration of the residency to dedicate to research, and when they most wished to undertake research. RESULTS: A total of 161 students (27.4%) responded. Overall, students preferred institutions with both clinical and research tracks rather than a single track for all residents, to delay their choice of track for as long as possible, and to do research during postgraduate year 4. Students indicated greatest interest in 3-6 months of research. Among those who self-identified as future academic researchers, interest in a research residency track was greater than among those who indicated the greatest interest in private practice positions. CONCLUSION: Many students are interested in performing research during their radiology residency. Interest in a research track is positively correlated with interest in a research academic career and negatively correlated with interest in private practice. PMID- 12529034 TI - Online evaluation in radiology residency programs. PMID- 12529035 TI - Effectiveness and student perceptions of standardized radiology clerkship lectures: a comparison between resident and attending radiologist performances. AB - RATIONALE AND OBJECTIVES: The authors' objective in this study was to determine whether radiology residents can effectively substitute for attending radiologists in delivering standardized lectures to medical students, without a decline in student performance on examinations or perception of the clerkship lectures. MATERIALS AND METHODS: Between September 2001 and July 2002, 40 3rd- and 4th-year medical students in the radiology clerkship at Rhode Island Hospital were given 11 standardized PowerPoint presentations. Half of the students were lectured by attending radiologists, and the other half were given the same lectures by radiology residents. At the end of each lecture, students completed surveys regarding their perceptions of the lecture. At the end of each clerkship, a written examination was administered to the students. The results of the surveys and the written examinations were then analyzed to determine whether there were significant differences between the perceptions and performances of these two student groups. RESULTS: There was no significant difference regarding the perceived overall lecture quality and no significant difference in final examination performance between the two student groups. Lectures by residents were perceived as having an environment in which students felt more comfortable asking questions. Attending radiologist lecturers, however, were perceived to have a greater ability to answer student questions, as well as to ask more appropriate questions of students. CONCLUSION: Radiology residents were as effective overall as attending radiologists at presenting standardized lecture material to medical students, with no difference in student perception of overall lecture quality or in student performance on examinations. PMID- 12529036 TI - Anti-DNA antibodies--overview of assays and clinical correlations. AB - Many authors have studied the links between levels of anti-dsDNA antibodies and disease activity in patients with SLE. Interpretation of these studies must take into account the facts that there are a range of possible assays for anti-dsDNA and a number of indices available for assessing disease activity. A recent study compared levels of various autoantibodies with organ specific disease activity assessed during the British Isles Lupus Assessment Group (BILAG) index. Anti dsDNA and anti-heparan sulphate levels were more likely to be raised in patients with renal than non-renal disease. Some anti-DNA antibodies are actually anti nucleosome antibodies, which lose DNA reactivity when purified under dissociating conditions. Patients with SLE have significantly increased levels of nucleosomes in their sera compared with healthy controls. In patients with SLE, reduced clearance of nucleosomes released from apoptotic cells may induce the formation of anti-nucleosome antibodies. PMID- 12529037 TI - Structural considerations of autoantibodies. AB - The mechanisms responsible for peptide-induced immunosuppression lupus-prone BWF1 mice were determined to be mediated via recognition by T cells, although the response was peptide-specific, as some accelerated the autoimmune response. Furthermore, this was associated with suppression of IFN-g and IL-4 in serum and increased TGF-b. Recent isolation of peptide-specific T cells should be helpful in sorting out the mechanisms responsible for these events. In separate studies, it was demonstrated that an anti-DNA antibody that enters cells is able to transport proteins linked to it, supporting the possibility that this system can be used as a therapeutic modality to modify specific cellular activities. PMID- 12529038 TI - Anti-DNA antibodies--structure and function. AB - Expression of monoclonal anti-DNA antibodies in vitro can be used to study the relationships between molecular structure, binding properties and pathogenicity. Bacterial and yeast systems can be used to produce antibody fragments such as Fab. The yields are potentially sufficient to allow structural studies such as crystallization, but purification of the anti-DNA Fab from the bacterial periplasm may be challenging. Mammalian cell expression systems produce lower yields, but the products are whole antibodies, which can be used in assays of pathogenicity. This article describes some recent experiments in which bacterial and mammalian systems were used to study human monoclonal anti-DNA antibodies. Light chain sequence motifs were found to be important both in binding to antigens and in determining pathogenicity of the antibodies in severe combined immunodeficiency mice. The distribution of B cell subpopulations is disturbed in patients with systemic lupus erythematosus (SLE). These patients, like those with infectious mononucleosis, have an overall B cell lymphopenia but an increased frequency of plasmablasts/early plasma cells in their blood. Some of these early plasma cells belong to clones that have rearranged the V(H) gene V4-34. There is a selective rise in immunoglobulins encoded by this gene in both infectious mononucleosis and SLE. PMID- 12529039 TI - Antigenic targets--workshop report. AB - Several excellent animal models for systemic lupus erythematosus are available to study the pathogenetic mechanisms of anti-DNA antibodies, immunological mechanisms for their origin and the influence of susceptibility genes for the development of anti-DNA antibodies. Furthermore several transgenic mice have been generated to study B cell tolerance against DNA. During the meeting recent progress on animal models for systemic lupus erythematosus was discussed. PMID- 12529040 TI - Cross-reactivity and pathogenicity of anti-DNA autoantibodies in systemic lupus erythematosus. AB - Autoantibodies to DNA were discovered over 40 years ago following the discovery a few years earlier of the 'LE' cell phenomenon by Hargraves and colleagues in 1948. These investigators noted that, when leucocytes were incubated with serum from lupus patients, changes in the nucleus could be seen together with phagocytosis of nuclear remnants by polymorphonuclear leucocytes. Since that time numerous studies in many laboratories have investigated almost every aspect of anti-DNA antibodies, partly to identify what determines their pathology. Whilst a subset of anti-DNA antibodies, especially anti-native, or double-stranded DNA (dsDNA) antibodies constitutes a hallmark of lupus disease and a diagnostic criterion, it is now clear that not all anti-DNA autoantibodies are of pathogenic relevance. Moreover, anti-DNA autoantibodies may also be found in other connective tissue disorders. Here we briefly review studies presented at the fifth international workshop on anti-DNA autoantibodies held in London to highlight relevant properties of pathogenic anti-DNA antibodies. PMID- 12529041 TI - Apoptosis and nucleosomes. AB - Drs Berden and Koutouzov presented evidence that nucleosomes are antigens in lupus pathogenesis and that apoptotic cells are the source of nucleosomes. Berden's group measured persistence of circulating nucleosomes and nucleosome antibody complexes in autoimmune mice and demonstrated nucleosomal deposition in skin of SLE patients as well as in renal lesions. Koutouzov reported that anti nucleosomes are among the earliest autoantibodies in MRL+/+ mice, appearing several weeks before anti-DNA antibodies. Treatment of the mice with a pro apoptotic drug, taxotere, accelerated autoantibody production and development of lesions. Herrmann proposed that persistent immunization results from reduced dead cell clearance and reduced production of immunosuppressive cytokines by defective scavenger macrophages. He also described accumulation of apoptotic cells in germinal follicles in SLE patients and attachment of nuclear antigens that are produced in apoptosis to the surface of follicular dendritic cells. Apoptosis derived nucleosomes may be important in both the immunizing and effector arms of pathogenesis. PMID- 12529042 TI - T cell-B cell interactions. AB - This session had as its central theme the analysis of peptide epitopes and their relationship with lupus pathogenesis. New information on the role of peptides opens up the possibility of treatments based on inducing immunological tolerance although care needs to be taken since it is difficult to predict flare or remission of disease after exposure to critical antigenic peptides. The provenance of these peptides may be self or foreign antigens, or autoantibody idiotypes. PMID- 12529043 TI - Workshop report on some new ideas about the treatment of systemic lupus erythematosus. AB - Our increased understanding of the pathogenesis of systemic lupus erythematosus is leading to new ideas about its therapy. In this session of the workshop the use of LJP 394 a B cell toleragen and the use of an anti-CD20 monoclonal antibody were discussed in some detail. Their rationale and early clinical results were reviewed; both have shown encouraging clinical and serological benefit. Definitive double-blind clinical trials are still, however, awaited. In addition, the intriguing notion of using a nasal instillation of a histone peptide was described and early work in an experimental model presented. PMID- 12529044 TI - Clinical laboratory assays for measuring anti-dsDNA antibodies. Where are we now? AB - It is over 40 years since anti-dsDNA antibodies were first described in patients with lupus. A wide variety of methods for detecting these immunoglobulins have been established. In this review, we consider the utility and practical problems that the main assays present. The clinical correlations between anti-dsDNA antibodies and clinical activity in patients with lupus is discussed. PMID- 12529045 TI - Many paths lead to lupus. PMID- 12529046 TI - Systematic analysis of sequences of anti-DNA antibodies--relevance to theories of origin and pathogenicity. AB - Sequence analysis of anti-DNA antibodies is important in determining the molecular features which distinguish potentially pathogenic antibodies from those which are less likely to be pathogenic. Previous analysis of murine anti-DNA antibody sequences suggested that particular murine immunoglobulin genes are used preferentially to encode such antibodies and that somatic mutations to arginine, asparagine and lysine may be important in the creation of DNA binding sites. In this paper, a systematic analysis of published human anti-DNA sequences shows no strong evidence for preferential usage of particular human V(H) or V(L) genes in anti-DNA antibodies. Somatic mutations in IgG and IgA antibodies are clustered in the complementarity determining regions (CDRs) due to the effect of antigen drive. This process contributes to an excess of arginine, asparagine and lysine residues in these CDRs, some of which are likely to play an important role in binding to DNA. Computer modeling and in-vitro expression experiments are likely to help define the roles played by these residues in antigen binding and pathogenicity more clearly. PMID- 12529047 TI - Molecular expression systems for anti-DNA antibodies--1. AB - Molecular expression systems can be used to produce whole antibodies or antibody fragments. The properties of these expression products can be tested in assays of binding or pathogenicity. Expression systems can be used to produce large quantities of antibodies which are already well-characterized, to produce new antibodies by repertoire cloning, or to produce slight modifications in the sequences of antibodies by mutagenesis prior to expression. This paper reviews the ways in which these methods have been used to study the structure and function of human and murine anti-DNA antibodies. A consistent finding, from experiments using a range of different expression methods and antibodies, is that sequence motifs including arginine residues play a major role in binding to DNA. These motifs can be present on either the heavy or the light chain, but are particularly reported in V(H)CDR3. PMID- 12529048 TI - Molecular expression systems for anti-DNA antibodies--2. AB - Antibodies to double-stranded DNA are the best-known serological markers of systemic lupus erythematosus, and are closely associated with its renal pathogenesis. How these antibodies recognize DNA is not fully understood. An understanding of the relationship between the functional attributes of an antibody with the three-dimensional structure of its antigen-combining site would allow an insight into the rules that dictate auto-antibody-nucleic acid interaction and consequent pathogenicity of the autoantibody. Data from such studies could assist the development of novel drugs as an approach to specific therapies that can inhibit or disrupt protein-nucleic acid interactions. A full understanding of the binding specificities can be achieved only by experimental determination of detailed three-dimensional structure of these antibodies alone, and of their complexes with specific DNA antigens. A prerequisite of such a study is the ability to produce multimilligram quantities of the antibody protein. However, these antibodies are particularly difficult to express, probably due to their DNA-binding activity. This review attempts to focus on the recent developments on the over-expression of anti-DNA antibody fragments in heterologous cell expression systems and their purification to homogeneity that would in turn allow their structural studies via crystallization. PMID- 12529049 TI - Transgenic models of tolerance and autoimmunity: with special reference to systemic lupus erythematosus. AB - Transgenic and knockout mouse carrying rearranged antigen-receptor genes have been invaluable for the elucidation of basic mechanisms in autoimmunity and have contributed new models of human autoimmune diseases. Several examples of transgenic models expressing rearranged immunoglobulin genes have been described. These models have provided a window into the events involved in this process, allowing the development and fate of self-reactive lymphocytes to be followed in vivo. In the B cell lineage, as in T cells, self-reactive cells have been found to undergo several distinct fates in vivo: they can be physically eliminated, functionally inactivated, or they can persist unchanged or become activated. Nevertheless the precise understanding of the molecular events leading to lymphocyte deletion, anergy or activation remains a challenge. PMID- 12529050 TI - Tumour necrosis factor alpha in systemic lupus erythematosus and anti-DNA autoantibody production. AB - Tumour necrosis factor (TNFalpha) is a cytokine with a wide range of diverse and at times paradoxical effects. These include immunoregulatory, lymphoid organogenesis and pro-inflammatory effects. In recent years, TNFalpha has become a focus of interest more for its inflammatory effects in a number of chronic autoimmune diseases. This interest culminated in the successful treatment of patients with rheumatoid arthritis, Crohn's diseases and ankylosing spondylitis with blocking antibodies or soluble TNFalpha receptors. Paradoxically, however, TNFalpha also has immunomodulatory effects in some autoimmune conditions such as lupus in some mouse models of the disease and in diabetes in the none-obese diabetic mouse. The role TNFalpha plays in human systemic lupus erythematosus is, however, controversial. In this article we review some of the studies carried out to elucidate the effects of TNFalpha in lupus disease and likely mechanisms of action. Further, we discuss recent data on the likely effects of blocking TNFalpha on anti-DNA autoantibody production. PMID- 12529051 TI - Triggers for anti-chromatin autoantibody production in SLE. AB - The formation of autoantibodies against chromatin is the main feature of systemic lupus erythematosis (SLE), an autoimmune disease, which is T-cell dependent and autoantigen-driven. Historically, antibodies against dsDNA, one of the components of chromatin, are considered as a hallmark of SLE. However, dsDNA is poorly immunogenic. Nucleosome-specific T helper cells have been identified. These T cells propagate not only nucleosome-specific antibodies, but also anti-dsDNA antibodies. Nucleosomes are formed during apoptosis by cleavage of chromatin, and evidence of disturbed apoptosis has been found especially in certain murine models of lupus. In addition to an increased rate of apoptosis, autoimmunity against chromatin might also result from an impaired phagocytosis of apoptotic material, for which strong evidence has been provided by studies in certain knock out mice (C1q, SAP, Dnase I). The induction of an immune response to nucleosomes could be enhanced by modifications of histones or DNA during apoptosis, altered presentation by antigen presenting cells or a viral infection. The release of nucleosomes and the formation of anti-chromatin autoantibodies result in formation of complexes, which bind to the glomerular basement membrane via heparan sulfate. This deposition incites glomerulonephritis, the most serious manifestation of SLE. PMID- 12529052 TI - Antigenic triggers and molecular targets for anti-double-stranded DNA antibodies. AB - While anti-double-stranded (ds)DNA antibodies are a characteristic serologic hallmark for SLE, the triggering antigen is unknown. Using phage display libraries, we identified DWEYSVWLSN as a peptide mimic of DNA for a pathogenic anti-dsDNA antibody. Peptide immunization of non-autoimmune mice induced anti dsDNA as well as other lupus-associated antibodies. Molecular analysis of the induced anti-dsDNA antibodies revealed several similarities with anti-dsDNA antibodies that appear spontaneously in lupus mice. Furthermore, lupus-prone mice immunized with this peptide DNA mimic had higher autoantibody titers as well as more severe nephritis. Anti-DNA antibodies may contribute to lupus nephritis via cross-reactivity with renal antigen. Using western blotting of lysates of mesangial cells from a lupus mouse, we found that a pathogenic anti-DNA antibody binds to alpha-actinin. High titers of anti-alpha-actinin antibodies were present in the sera and kidney eluates of lupus mice with active disease. Binding to alpha-actinin was diminished in mesangial cells derived from BALB/c mice, suggesting that target antigen expression may play a role in determining autoantibody binding to the kidney. We conclude that a pathogenic, lupus-like autoantibody response can be induced by a peptide antigen, and that alpha-actinin is a cross-reactive renal target for the pathogenic anti-dsDNA autoantibody response in lupus mice. PMID- 12529053 TI - Disturbances in peripheral blood B cell subpopulations in autoimmune patients. AB - A variety of cell surface markers are being used to identify B cell subpopulations in peripheral blood. Currently at least eight subpopulations have been identified. Analyses of healthy individuals indicate that in general the various B cell subpopulations exist in relatively similar ratios in unrelated individuals. It has been demonstrated that B lymphocyte homeostasis is disturbed during infection and autoimmune disease. In this review we compare the distribution of B cell subpopulations in the peripheral blood of patients with systemic lupus erythematosus, rheumatoid arthritis and primary Sjogren's syndrome with each other, and with healthy individuals. The different autoimmune diseases have distinct changes in the B cell subpopulations. Understanding the nature of these B subpopulation signatures will potentially impact understanding the mechanisms of disease, diagnosis and therapy. PMID- 12529054 TI - The role of VH determinants in systemic lupus erythematosus. AB - The V-regions of anti-DNA antibodies contain determinants which can drive the autoimmune in SLE. Most of the evidence comes from murine studies where VH derived epitopes accelerate the disease process in lupus prone-mice and can elicit mild inflammatory changes reminiscent of lupus in healthy animals. T helper cells reactive with VH peptides arise spontaneously during the disease and are thought to assist production of both anti-peptide antibodies and the generation of autoantibodies that deposit in the glomeruli. In mice stimulatory epitopes may be unique to autoantibodies. As tolerogens VH peptides may delay or diminish the autoimmune response by altering the production of cytokines. An artificial VH peptide, (pCONCENSUS) has been derived and this inhibits responses to VH and other autoantigens but leaves the murine immune system intact and able to generate reponses to external antigens. Limited number of studies of V-region determinants of human anti-DNA MAbs indicate prior sensitization of lupus T cells to VH determinants and that V-region reactive T cells are not deleted in periphery of healthy individuals. PMID- 12529055 TI - Lowering anti-dsDNA antibodies--what's new? AB - Antibodies to dsDNA are specific to SLE and are pathogenic, both due to their ability to deposit in tissues through a variety of mechanisms, and to their ability, when present in immune complexes, to activate inflammatory cells. The relationship of serum anti-dsDNA antibody levels to disease activity is a complex one and the factors that determine whether or not such antibodies will be pathogenic in an individual SLE patient are incompletely understood. Although anti-dsDNA antibodies can be made by naive B cells and B cells belonging to the B1 and marginal zone subsets, pathogenic anti-dsDNA antibodies have the hallmarks of germinal center development and exposure to T cell help, including accumulation of somatic mutations and class switching to the IgG isotype. Epitope spreading may result in aquisition of cross-reactivities with multiple target organ antigens and aquisition of a memory phenotype will allow these B cells to acquire antigen presentation functions that amplify the autoreactive response. In the early stages of disease, or after remission induction protocols, autoreactive B cells may be susceptible to treatments that target T cell costimulation or that deplete or tolerize naive and mature B cells. Therapeutic approaches targeting innate immune responses or regulatory T cells are starting to be tested in pre clinical models. In later disease stages, memory and plasma cell accumulation may render patients more resistant to this type of therapeutic approach. Deposition of anti-dsDNA antibodies in target tissues can stimulate an inflammatory cascade that leads to tissue damage. A number of murine models have now been developed that show that interruption of this cascade can prevent or reverse such damage. This type of approach may be beneficial for individuals with established disease. As we learn more about the specific defects that cause SLE, it may become possible to individualize therapy based on patient specific biologic markers. PMID- 12529056 TI - Dynamical evolutionary psychology: individual decision rules and emergent social norms. AB - A new theory integrating evolutionary and dynamical approaches is proposed. Following evolutionary models, psychological mechanisms are conceived as conditional decision rules designed to address fundamental problems confronted by human ancestors, with qualitatively different decision rules serving different problem domains and individual differences in decision rules as a function of adaptive and random variation. Following dynamical models, decision mechanisms within individuals are assumed to unfold in dynamic interplay with decision mechanisms of others in social networks. Decision mechanisms in different domains have different dynamic outcomes and lead to different sociospatial geometries. Three series of simulations examining trade-offs in cooperation and mating decisions illustratehow individual decision mechanisms and group dynamics mutually constrain one another, and offer insights about gene-culture interactions. PMID- 12529057 TI - Attention, spatial representation, and visual neglect: simulating emergent attention and spatial memory in the selective attention for identification model (SAIM). AB - The selective attention for identification model (SAIM) is presented. This uses a spatial window to select visual information for recognition, binding parts to objects and generating translation-invariant recognition. The model provides a qualitative account of both normal and disordered attention. Simulations of normal attention demonstrate 2-object costs and effects of object familiarity on selection, global precedence, spatial cueing, and inhibition of return. When lesioned, SAIM demonstrated either view- or object-centered neglect or spatial extinction, depending on the type and extent of lesion. The model provides a framework to unify (a) object- and space-based theories of normal selection, (b) dissociations within the syndrome of unilateral neglect, and (c) attentional and representational accounts of neglect. PMID- 12529058 TI - Goal-referenced selection of verbal action: modeling attentional control in the Stroop task. AB - This article presents a new account of the color-word Stroop phenomenon (J. R. Stroop, 1935) based on an implemented model of word production, WEAVER++ (W. J. M. Levelt, A. Roelofs, & A. S. Meyer, 1999b; A. Roelofs, 1992, 1997c). Stroop effects are claimed to arise from processing interactions within the language production architecture and explicit goal-referenced control. WEAVER++ successfully simulates 16 classic data sets, mostly taken from the review by C. M. MacLeod (1991), including incongruency, congruency, reverse-Stroop, response set, semantic-gradient, time-course, stimulus, spatial, multiple-task, manual, bilingual, training, age, and pathological effects. Three new experiments tested the account against alternative explanations. It is shown that WEAVER++ offers a more satisfactory account of the data than other models. PMID- 12529059 TI - Interval timing as an emergent learning property. AB - Interval timing in operant conditioning is the learned covariation of a temporal dependent measure such as wait time with a temporal independent variable such as fixed-interval duration. The dominant theories of interval timing all incorporate an explicit internal clock, or "pacemaker," despite its lack of independent evidence. The authors propose an alternative, pacemaker-free view that demonstrates that temporal discrimination can be explained by using only 2 assumptions: (a) variation and selection of responses through competition between reinforced behavior and all other, elicited, behaviors and (b) modulation of the strength of response competition by the memory for recent reinforcement. The model departs radically from existing timing models: It shows that temporal learning can emerge from a simple dynamic process that lacks a periodic time reference such as a pacemaker. PMID- 12529060 TI - Core affect and the psychological construction of emotion. AB - At the heart of emotion, mood, and any other emotionally charged event are states experienced as simply feeling good or bad, energized or enervated. These states- called core affect--influence reflexes, perception, cognition, and behavior and are influenced by many causes internal and external, but people have no direct access to these causal connections. Core affect can therefore be experienced as free-floating (mood) or can be attributed to some cause (and thereby begin an emotional episode). These basic processes spawn a broad framework that includes perception of the core-affect-altering properties of stimuli, motives, empathy, emotional meta-experience, and affect versus emotion regulation; it accounts for prototypical emotional episodes, such as fear and anger, as core affect attributed to something plus various nonemotional processes. PMID- 12529061 TI - What does sexual orientation orient? A biobehavioral model distinguishing romantic love and sexual desire. AB - Although it is typically presumed that heterosexual individuals only fall in love with other-gender partners and gay-lesbian individuals only fall in love with same-gender partners, this is not always so. The author develops a biobehavioral model of love and desire to explain why. The model specifies that (a) the evolved processes underlying sexual desire and affectional bonding are functionally independent; (b) the processes underlying affectional bonding are not intrinsically oriented toward other-gender or same-gender partners: (c) the biobehavioral links between love and desire are bidirectional, particularly among women. These claims are supported by social-psychological, historical, and cross cultural research on human love and sexuality as well as by evidence regarding the evolved biobehavioral mechanisms underlying mammalian mating and social bonding. PMID- 12529062 TI - The intelligence of the moral intuitions: comment on Haidt (2001). AB - The social intuitionist model (J. Haidt, 2001) posits that fast and automatic intuitions are the primary source of moral judgments. Conscious deliberations play little causal role; they are used mostly to construct post hoc justifications for judgments that have already occurred. In this article, the authors present evidence that fast and automatic moral intuitions are actually shaped and informed by prior reasoning. More generally, there is considerable evidence from outside the laboratory that people actively engage in reasoning when faced with real-world moral dilemmas. Together, these facts limit the strong claims of the social intuitionist model concerning the irrelevance of conscious deliberation. PMID- 12529063 TI - Permission to cross the border: effective policy reduces high-risk drinking by Marines. AB - OBJECTIVE: At a Marine Corps base near the Mexican border, off-base alcohol- related incidents increased when serving alcohol to personnel under age 21 on the base was no longer permitted. The bars across the Mexican border, where the legal drinking age is 18, are an important source of alcohol for the Marine enlisted men. To reduce cross-border drinking, the base command instituted a "chit" system requiring enlisted men to receive written permission to cross the border. This study evaluates the effectiveness of that policy. METHOD: Breath test surveys of American youths returning from Tijuana to San Diego County provided counts and blood alcohol concentration (BAC) of marines returning from a night of drinking in Mexico before and after the "chit" policy. Comparison counts and breath tests for Navy personnel from a San Diego base not affected by the "chit" policy were available for the same time periods. RESULTS: The number of underage marines returning from Mexican bars was reduced by 78%, and of those the number with illegal (> or = .08%) BACs was reduced by 84%. There was no significant change for the comparison group of sailors. CONCLUSIONS: It appears that the "chit" policy was highly effective at least for the 6-month post-period covered by this study. This apparent success influenced the implementation of the "chit" policy on all naval facilities in the San Diego area. PMID- 12529064 TI - Treating alcohol problems with self-help materials: a population study. AB - OBJECTIVE: An experimental trial was used to assess the effectiveness of a self help book and a personalized assessment-feedback intervention, both separately and in combination with each other, in a general population survey. METHOD: Participants (N = 86; 66.3% male) were recruited through a random digit dialing telephone survey conducted by the Centre for Addiction and Mental Health, Toronto, Ontario, Canada. Respondents were randomly assigned to one of four conditions in a two-by-two factorial design: "no-intervention" control group, "personalized feedback only," "self-help book only" and "both personalized feedback and self-help book." Respondents were followed up in 6 months' time, and differences in drinking status were compared between experimental conditions using a multivariate analysis of covariance with baseline drinking severity as the covariate. RESULTS: Support was provided for an interaction hypothesis in which respondents who received both interventions reported significantly improved drinking outcomes at 6-month follow-up, compared with respondents who received just one of the interventions or who received no intervention. CONCLUSIONS: Because respondents were recruited from a representative sample of the general population into a randomized trial with a no-intervention control group, this research design maximized both external and internal validity in examining the effectiveness of self-help interventions. PMID- 12529065 TI - Enhancing the effectiveness of traditional interventions with drinking drivers by adding brief individual intervention components. AB - OBJECTIVE: The purpose of the research was to determine whether the effectiveness of a traditional DUI (Driving Under the Influence) group intervention program could be enhanced by the addition of two brief individual intervention sessions and a follow-up. The differential effectiveness of the individual intervention component was examined for four offender subgroups (young minorities, problem drinkers, women and depressed offenders) that had been previously identified as at high risk or vulnerable. METHOD: Adjudicated first DUI offenders (N = 4,074), of whom 776 (19%) were female, were randomly assigned to a standard first offender program or an enhanced standard program that included two short individual sessions and a brief follow-up session. The setting was a mandated first-offender program in 10 Mississippi locations. RESULTS: Depressed offenders who were assigned to the enhanced program were 35% less likely to recidivate than those assigned to the standard program. The effectiveness of the two programs did not differ significantly for offenders who self-reported low depression. No significant interaction effects were found between program type and age, minority status or gender. After depressed mood was controlled for, problem-drinker status was not related to program effectiveness; however, problem drinkers had higher depression rates. CONCLUSIONS: Results suggest that the combination of a standard first-offender program with brief individual counseling can be effective for DUI offenders who report depressed mood and who are at high risk for recidivism. A five-item screen for sad/depressed mood from a widely used DUI risk assessment instrument identified offenders who benefited from the enhanced intervention. PMID- 12529066 TI - Examining mechanisms of action in 12-step treatment: the role of 12-step cognitions. AB - OBJECTIVE: Twelve-step treatment appears to be modestly effective in treating alcohol and cocaine dependence, but little is known about its mechanisms of action. The primary aim of this study was to examine the hypothesized mechanism of action of 12-step cognitions. In addition, the psychometric properties of a measure used to assess 12-step cognitions were tested. METHOD: The psychometric properties of a measure to assess 12-step cognitions were examined in a sample of individuals (N = 370, 61% male) seeking inpatient (n = 118) or outpatient (n = 252) 12-step-oriented community treatment. Additional analyses of data from the inpatient sample examined changes in cognitions from entry to discharge and 12 month follow-up. The univariate and multivariate relationship of cognitions at discharge, and substance use outcomes across the 12-month follow-up were also examined in the inpatient sample. RESULTS: The measure of 12-step cognitions demonstrated good internal consistency. Findings supported some, but not all, of the hypothesized active ingredients of 12-step treatment. Cognitive processes common to many treatment-approaches significantly mediated outcomes, whereas processes unique to the 12-step approach (e.g., spirituality) did not. Overall, some 12-step cognitions at discharge were good predictors of outcome, explaining as much variance in outcome as illness severity factors (e.g., dependence severity). CONCLUSIONS: Results provided greater support for 12-step treatment theory than those reported in prior studies. At the same time, findings suggest that diverse treatment approaches may operate through a common set of active ingredients. PMID- 12529067 TI - Who goes to alcohol and drug treatment? Understanding utilization within the context of insurance. AB - OBJECTIVE: This study assesses the differential roles of demographic characteristics, problem severity, motivational factors and insurance in entrance into alcohol and drug treatment. METHOD: A probability sample of adult problem and dependent drinkers from a Northem California county (N = 672) and consecutive adult intakes from the county's public (n = 298) and private (n = 628) alcohol and drug treatment programs. Interviewers conducted telephone screening for individuals meeting problem-drinking criteria in the general population; in person interviews were conducted for those who screened positive, and also for individuals entering treatment. RESULTS: Insurance did not play an independent role in treatment entry. In logistic regression analysis, using a model from the medical utilization literature, black ethnicity (odds ratio [OR] = 2.98, p < .001); older age (OR = 4.67, p < .001); less education (OR = 1.81, p < .01); legal (OR = 7.46, p < .001) and work (OR = 3.57,p <.001) pressures; higher psychiatric (OR = 4.03,p <.001) and drug (OR = 3.04,p <.001) severity; and social consequences (OR = 1.35, p < .01) predicted treatment entry. Interventions from legal sources (OR = 6.22, p < .01) were related to entering treatment for the uninsured; legal (OR = 7.02, p < .001), workplace (OR = 6.01, p < .001) and family (OR = 1.62, p < .05) pressures were related for the insured. CONCLUSIONS: Social consequences, rather than severity of dependence, in both insured and uninsured problem drinkers are important predictors of treatment entry when traditional utilization measures are controlled. Public programs play a crucial part in the alcohol and drug treatment system, and their viability is important as health policy changes. PMID- 12529068 TI - Physical and sexual abuse history and addiction treatment outcomes. AB - OBJECTIVE: Prior research on patients with substance use disorders has shown that lifetime physical or sexual abuse is associated with more impaired functioning at treatment intake. The present study sought to determine whether physical or sexual abuse also predicted treatment response (posttreatment outcomes) of individuals with substance use disorders. METHOD: Male (n = 19,989) and female (n = 622) veterans with substance use disorders were assessed with the Addiction Severity Index (ASI) early in treatment and reassessed an average of 12 months later. Treatment outcomes were compared for patients who did and did not report prior physical or sexual abuse in the initial ASI interview. RESULTS: Lifetime physical or sexual abuse predicted worse outcomes in six of seven domains of functioning, after controlling for baseline functioning, psychiatric diagnoses and demographic variables. Although women were more likely than men to report being abused, the effect of abuse on treatment outcomes was similar for both genders. Psychiatric problems at baseline mediated the effect of abuse history on outcomes. Abuse history moderated the effect of treatment intensity (contacts per month) on outcomes: More frequent treatment contacts were more beneficial for abused patients than for nonabused patients. CONCLUSIONS: Individuals with substance use disorders who have a history of physical or sexual abuse may have higher risk for problematic treatment outcomes as a result of greater psychiatric problems, deficits in social support and possible difficulties in establishing treatment alliance. Clinicians may consider increasing the duration and intensity of treatment to temper the negative effects of abuse on later functioning. PMID- 12529069 TI - Alcohol's direct and indirect effects on men's self-reported sexual aggression likelihood. AB - OBJECTIVE: This study investigated pathways through which alcohol's direct and indirect expectancy effects and direct physiological effects influenced men's self-reported sexual aggression likelihood after they read a violent pornographic story. The indirect effects of participants' affective responses and cognitive judgments of story characters were also examined. METHOD: Male social drinkers (N = 135), recruited through newspaper ads in a large western city, were randomly assigned to one of three beverage conditions: alcohol, placebo or control. After completing pretest measures, subjects read a violent pornographic story and reported their sexual arousal, affect, cognitive judgments and sexual aggression likelihood. RESULTS: Pre-existing expectancies operated directly and interactively with alcohol consumption on reported sexual aggression likelihood. The influence of expectancies on sexual aggression likelihood also occurred indirectly through positive affect and cognitive judgments of assailant force and victim enjoyment. Situational consumption effects were influenced by cognitive judgments. The expectation of receiving alcohol indirectly affected sexual aggression likelihood through its effect on perception of the assailant's typicality. CONCLUSIONS: Among men who have contact with violent pornography, alcohol can have both direct and indirect effects on reported sexual aggression likelihood. In addition to the presence of situational myopia and expectancy effects, pre-existing expectancies can play a significant role both alone and interactively in affecting this tendency. PMID- 12529070 TI - Alcohol-related aggression in men and women: the influence of dispositional aggressivity. AB - OBJECTIVE: The purpose of this study was to investigate the influence of dispositional aggressivity on the alcohol-aggression relation in men and women. METHOD: Subjects were 204 healthy social drinkers (111 men) between 21 and 35 years of age. Dispositional aggressivity was measured using the Buss-Perry Aggression Questionnaire. Following the consumption of either an alcohol or placebo beverage, subjects were tested on a modified version of the Taylor Aggression Paradigm, in which electric shocks are received from and administered to a fictitious opponent during a competitive task. Aggression was operationalized as the shock intensities administered to the fictitious opponent under conditions of low and high provocation. RESULTS: Of all the variables, provocation was the strongest elicitor of aggression. Overall, persons with high dispositional aggressivity exhibited more aggression than did those with low dispositional aggressivity. Alcohol increased aggression for persons with high, but not for those with low, dispositional aggressivity. This effect was stronger under conditions of low provocation. Furthermore, men and women with low dispositional aggressivity did not differ in aggression. Men with high dispositional aggressivity, however, were more aggressive than their female counterparts. CONCLUSIONS: This is the first investigation to examine the influence of dispositional aggressivity on the alcohol-aggression relation in men and women. The results highlight the fact that alcohol consumption does not increase aggression in all persons and in all situations. An important goal for future research is to identify which individual difference and contextual factors are most important in determining who will, and will not, behave in an aggressive manner when intoxicated. PMID- 12529071 TI - Psychiatric comorbidity, continuing care and mutual help as predictors of five year remission from substance use disorders. AB - OBJECTIVE: In a cohort of 2,595 male patients in VA intensive treatment programs for substance use disorders (SUD), we tested whether psychiatric comorbidity, outpatient care and mutual help group attendance during the first two follow-up years predicted remission status at Year 5, controlling for covariates. METHOD: Logistic regression modeling of longitudinal data was used to test the hypotheses. RESULTS: Dual diagnosis patients were less likely to be in remission at Year 5 than SUD-only patients. Outpatient care was at best only weakly related to Year 5 remission status. By contrast, mutual help involvement substantially improved the chances of substance use remission at Year 5 for both SUD-only and dual diagnosis patients. Mutual help involvement did not, however, offset the poorer prognosis for dual diagnosis patients. CONCLUSIONS: Because mutual help groups specifically targeted to individuals with comorbid substance use and psychiatric disorders are currently rare, further research is recommended to investigate whether they are more effective than standard SUD mutual help groups in facilitating the recovery of persons with dual diagnoses. PMID- 12529072 TI - Predictors of contact with public service sectors among homeless adults with and without alcohol and other drug disorders. AB - OBJECTIVE: Homeless persons with alcohol and other drug (AOD) disorders face multiple problems that go beyond their AOD use. As a consequence, they commonly access services in multiple sectors in addition to the AOD treatment system. This study examined the predictors of contact with agencies in the health, mental health, social welfare and criminal justice sectors by AOD status among a probability sample of homeless adults in Houston, Texas. METHOD: Cross-sectional data were collected from a multistage random sample of 797 homeless adults (579 men), age 18 or older, who were living in shelters and on the streets of Houston in 1996. Structured face-to-face interviews provided screening diagnoses for AOD disorders, self-report data on AOD treatment use and candidate predictors of treatment use. Service use was tracked retrospectively through administrative data obtained from 10 federal, state, county and municipal agencies that provide finding for physical and mental health services and AOD treatment, as well as emergency income; we also tracked criminal justice contacts. Logistic regression analyses were stratified by AOD status. RESULTS: Adjusting for eligibility factors, key aspects of need were significant predictors of any utilization among those without an AOD problem, but not for those with an AOD problem. For those with AOD disorders, contact with one sector was not predictive of contact with other sectors. CONCLUSIONS: Our findings indicate that AOD disorders hinder utilization of public sector services by homeless persons. These disorders may be masking need or otherwise acting as a barrier to accessing treatment and support. PMID- 12529073 TI - Attendance and alcohol use at parties and bars in college: a national survey of current drinkers. AB - OBJECTIVE: This study examines attendance and alcohol use at parties and bars among college students by gender, residence, year in school and legal drinking age. METHOD: The study participants were respondents in the 1997 and 1999 Harvard School of Public Health College Alcohol Study (CAS). The combined sample consisted of 12,830 students (61% women) who reported use of alcohol in the past 30 days prior to interview. Their responses provided information on attendance and alcohol use at parties (dormitory, fraternity, off campus) and off-campus bars. Logistic regression analyses examined the influence of gender, residence, year in school and legal drinking age related to attendance, drinking/non drinking and heavy drinking (5 or more drinks) at each select setting. RESULTS: Consistent with the literature, fraternity/ sorority parties were occasions of heavy drinking (49%) among drinkers in those settings, yet they drew upon smaller proportions of students (36%) when compared to off-campus parties (75%) and off campus bars (68%). Off-campus parties (45%) and bars (37%) were also occasions for heavy drinking among drinkers in these settings. College residence was shown to relate to differential exposure to drinking settings, but residence had less impact on the decision to drink and the level of heavy drinking. Attendance at parties decreased with advance in school years, but attendance at off-campus bars increased. Although heavy drinking at off-campus bars decreased with advancing grade year in school, slightly higher proportions of under-age students (41%) compared to students of legal drinking age (35%) exhibited heavy drinking at off campus bars. CONCLUSIONS: The identification of high-risk settings and their correlates serves to better understand the development of heavy drinking on college campuses. Off-campus parties, as compared to campus parties and bars, may pose greater difficulties related to successful intervention. PMID- 12529074 TI - Recent drinking history: association with family history of alcoholism and the acute response to alcohol during a 60 mg% clamp. AB - OBJECTIVE: Family history of alcoholism (FHA) is associated with increased drinking history, which can be a confounding factor in studies of the influence of FHA on the acute response to alcohol. The objective of this analysis was to investigate the association between recent drinking history (RDH) and FHA in a sample of family history positive (FHP; n = 55, 28 women) and family history negative (FHN; n = 55, 29 women) subjects, and to explore the influence of RDH on the response to alcohol during a 60 mg% clamp. METHOD: RDH was measured using daily diary and timeline followback methods. The total number of drinks in the 4 week (TD28) and 1-week (TD07) intervals prior to the study were determined, as well as the number of drinking days in the same intervals. Dependent measures of brain function were obtained at baseline (B0), immediately after the target BrAC was achieved (B1) and 105 minutes later (B2). The alcohol response was quantified as an initial response (ira = B1-B0) and an adaptive response (ada = B2-B1). The association between RDH and the ira and ada measures was tested using multivariate regression. RESULTS: The RDH measures showed a large variance across subjects, with no significant differences between FHP and FHN groups in the study sample. The initial responses for subjective perceptions of "high" and "intoxicated," Alcohol Sensation Scale scores and scores for the grooved pegboardtask were significantly negatively associated with TD28. Acute tolerance to perceptions of "high" and "intoxication" was significantly negatively associated with TD28. CONCLUSIONS: Heavy drinking history is associated with a decreased initial response to alcohol and greater acute tolerance to alcohol, particularly for subjective measures. Although RDH was not associated with FHA in this study, it may be an important determinant of the response to alcohol. PMID- 12529075 TI - Does caffeine counteract alcohol-induced impairment? The ironic effects of expectancy. AB - OBJECTIVE: Studies have shown that expectations of alcohol-induced impairment can produce adaptive responses to alcohol that reduce the degree of behavioral impairment displayed. The present study tested psychomotor performance following combined caffeine and alcohol administration in 42 social drinkers (23 men). Subjects were led to expect either that caffeine would antagonize alcohol-induced impairment or that it would have no effect. The study tested the hypothesis that drinkers who expected an antagonist effect of caffeine would display greater alcohol impairment than those who expected no antagonist effect. METHOD: Groups practiced a pursuit rotor task and received a moderate dose of alcohol (0.65 g/kg) combined with either 4.0 mg/kg caffeine or placebo caffeine. Some groups were led to expect that caffeine would counteract the impairing effect of alcohol and others were led to expect no counteracting effect. Psychomotor performance was then tested over a 3-hour period. RESULTS: In accord with the hypothesis, groups led to expect counteracting effects of caffeine displayed greater impairment than those led to expect no counteraction. Caffeine had no significant antagonist effect on alcohol impairment. CONCLUSIONS: The findings suggest that compensation for alcohol impairment occurs when drinkers hold clear expectations that the drug will disrupt performance. When no such clear expectation exists, no compensatory response occurs and the impairing effects of alcohol are observed. PMID- 12529076 TI - Mode effects for collecting alcohol and other drug use data: Web and U.S. mail. AB - OBJECTIVE: The present study examined mode effects for collecting alcohol and other drug use data using a Web-based survey mode and a U.S. mail-based survey mode for comparison. METHOD: A survey regarding alcohol and other drugs was administered to a randomly selected sample of 7,000 undergraduate students attending a large midwestern research university in the spring of 2001. The sample was randomly assigned to either a Web-based survey mode (n = 3,500) or a U.S. mail-based mode (n = 3,500). RESULTS: The Web survey mode of administration resulted in a final sample that more closely matched the target sample in gender mix than did the U.S. mail survey mode. The response rate for the Web survey mode was significantly higher than for the U.S. mail survey mode. Chi-square results indicated there were significant differences in response propensity by several sample characteristics including sex, race, class year and academic credit hours. Multivariate logistic regression results revealed significant racial and gender differences in the response propensity between and withinmodes. After controlling for design discrepancies, there were no significant differences between modes in data quality or substantive responses to substance-use variables. CONCLUSIONS: The findings of the present study provide strong evidence that Web surveys can be used as an effective mode for collecting alcohol and other drug use data among certain populations who have access to the Internet and high rates of use. Web surveys provide promise for enhancing survey research methodology among undergraduate college students. PMID- 12529077 TI - Are retrospectively reconstructed blood alcohol concentrations accurate? Preliminary results from a field study. AB - OBJECTIVE: The purpose of this study was to evaluate the relationship between blood alcohol concentration (BAC) derived from an in vivo breath test and retrospective estimates of BAC (eBAC) for the same drinking event. The relationship was expected to be lower at higher levels of BAC as a result of inaccuracies in the recall of self-report data. METHOD: The sample consisted of 44 persons (64% male) who were walking away from a bar district adjacent to a university campus. After giving informed consent, participants provided a breath sample and contact information. The next day a research assistant telephoned the participants and conducted a brief interview, using the equation prescribed by the National Highway Traffic Safety Administration, to obtain the self-report data necessary to calculate the eBAC for the previous evening. RESULTS: The actual BACs and eBACs were not significantly different, and they were highly correlated (r = 0.84, n = 43). BAC did predict the magnitude of discrepancy between the two, and at BACs over 0.08, the relationship was nonsignificant. CONCLUSIONS: Although self-report data can be used to approximate the BAC obtained during a naturally occurring drinking event the relationship is moderated by measured level of intoxication. Retrospective calculations are less accurate when estimating higher BACs. PMID- 12529078 TI - Effects of alcohol misuse on gambling patterns in youth. AB - OBJECTIVE: Gambling and alcohol misuse are prevalent among youth and may be part of a common problem behavior syndrome. It was hypothesized that alcohol misuse would predict a pattern of increased youth gambling or a pattern of stable high gambling after controlling for key sociodemographic, socialization and individual factors. METHOD: Data were analyzed from two longitudinal studies of youth living in a western New York metropolitan area. Respondents' gambling at two times over the course of 12-18 months was classified into one of five gambling pattern groups, representing flat-low, increasing, flat-medium, flat-high and decreasing levels of gambling. RESULTS: Alcohol misuse among males predicted increasing gambling over time or a pattern of stability of high rates of gambling even after controlling for socioeconomic status, race, age, impulsivity and parental monitoring in the family study. Higher parental monitoring and lower alcohol misuse were significant in predicting a decreasing pattern of gambling among males in the male delinquency study. For females in the family study, alcohol misuse predicted an increasing pattern of gambling only when other factors such as high impulsivity or low parental monitoring were present. CONCLUSIONS: An understanding of adolescent gambling must take into account a variety of demographic, socialization and individual factors, as well as the co-occurrence of alcohol misuse. PMID- 12529079 TI - Nursing within Australia is currently the focus of attention at both state and national levels. PMID- 12529080 TI - Nursing career structures: issues and implications. PMID- 12529081 TI - The craft of teaching qualitative research: linking methodology to practice. AB - Qualitative inquiry provides a means of apprehending the personal and contextual experiences of health and health care as well as the complex social, cultural and political issues that influence services and professional practice. Despite its usefulness in informing clinical practice, it is undervalued and under-utilised in contemporary nursing research. A number of organisational, ideological and educational reasons exist for this situation. This paper addresses educational issues, specifically teaching practice. We discuss craft knowledge, an important but often obscured practice, and its use in qualitative research education, drawing on the findings of a project that investigated a group of nursing academics' experiences and perceptions about teaching qualitative research. We argue that craft knowledge offers teachers a means to inspire and engage clinician students to learn not only the content and research skills required to do good qualitative research but to learn more about the standpoint and sensibilities of being qualitative researchers. In this way craft knowledge does two things. First it helps to move beyond the content versus process polemic that often dominates educational debate. Second, by enhancing the quality of educational experience, it potentially helps clinicians to value qualitative inquiry and thus defend and use it to inform clinical practice. PMID- 12529082 TI - Discharge planning from the acute sector for people over the age of 65. AB - This paper reports the findings relating to discharge planning from a larger study (Hegney et al 2001) that aimed to investigate options to improve care co ordination between the aged, community, acute and primary care sectors for the elderly residents of a large provincial town. The data, which were collected from a wide variety of aged care stakeholders in the region, are consistent with previous Australian studies. First, it appears that inappropriate discharge from acute hospitals occurs in Toowoomba because there is a perceived lack of information regarding the available community care options and a lack of suitable models to support people's care needs within their own homes. Second, inappropriate discharge practices from acute hospitals occur due to a lack of knowledge and communication on the part of health professionals employed in these settings with regard to the on-going health care needs of elderly people. Third, there is little discharge planning occurring that includes the general practitioner in post-discharge care. Finally, inappropriate discharge can also be attributed to the lack of sub-acute/step-down/rehabilitation options and long waiting lists for community home care services in the region. PMID- 12529083 TI - Silence--not so golden? AB - In remote Australia, when the dust settles after a windstorm, nothing remains the same--everything becomes coated with the dust that penetrates all crevices and surfaces of life. So it is with remote area nursing--from the early 1980s until today, remote area nurse voices have stirred up the winds of change. This followed over almost two centuries of silence. Token support came from local community leaders and health ministers to strengthen remote area nursing voices. But why has it taken almost two centuries? This paper represents the results of my interaction with the public discourses of Australian nurses during the nineteenth century (when there were barely any nurses at all) and with remote area nurses of the twentieth century. This led to the development of a model to explore the process of agency and breaking silence. It became evident that within the ranks of nursing itself, a process of downward closure was enacted. A strategy of dual closure was enacted by nurses against their own colleagues at an intra-occupational level and was operationalised by legalistic (State registrations) and credentialist (exercising control over content and standards) tactics. PMID- 12529084 TI - Missed opportunities in the field: caring for clients with co-morbidity problems. AB - This discussion paper presents key issues associated with caring for people with concurrent alcohol or drug and mental health problems. By way of anecdotal stories and personal experiences, it draws attention to 'missed opportunities' for effective care of clients within a general hospital and community mental health service. As educators, inicians and students, we are concerned our own attempts to improve the care of these clients may have been less than successful due to various factors, and believe such information is important to share with the wider nursing profession. We hope the paper will stimulate interest amongst general, mental health and drug and alcohol nurses, and researchers and educators to better address the issue of alcohol, drug and mental health co-morbidity in our community. PMID- 12529085 TI - Making the transition from enrolled to registered nurse. AB - Each year among the university graduates who become eligible to register as nurses for the first time, there are a number of people who have previously trained and practiced as enrolled nurses. There is little research that examines the experiences and needs of this unique group of simultaneously experienced and novice nurses. In this paper, a phenomenological approach was used to examine the lived experience of nine former enrolled nurses making the transition from nursing student to graduate nurse. Data from extensive interviews with these nine nurses was examined using van Manen's (1990) steps of thematic analysis. The grouping of concepts into subthemes led to the identification of three main themes that described the nurses' experiences of transition: responsibility and accountability, change, and job satisfaction. Excerpts from interview transcripts are used as exemplars in discussion of the meaning and importance of these findings. The findings of this study will inform nurse education and graduate transition support services. PMID- 12529086 TI - Vancomycin resistant enterococci: evidence based practice to reduce transmission. PMID- 12529087 TI - Surveillance for traumatic brain injury deaths--United States, 1989-1998. AB - PROBLEM/CONDITION: Data indicate that approximately 50,000 U.S. residents die as a result of traumatic brain injury (TBI) annually. Survivors of TBI are often left with neuropsychologic impairments that result in disabilities affecting work or social activity. During 1979-1992, TBI-related death rates declined 22%, from 24.6 to 19.3 deaths/100,000 population. This report describes the epidemiology and trends in TBI-related mortality during 1989-1998. REPORTING PERIOD: January 1, 1989-December 31, 1998. DESCRIPTION OF SYSTEMS: The National Center for Health Statistics (NCHS) Multiple Cause of Death public use data were analyzed for this study. RESULTS: During 1989-1998, an annual average of 53,288 deaths (range: 51,848-54,501) among U.S. residents were associated with TBI. TBI-related death rates declined 11.4%, from 21.9 to 19.4/100,000 population. The major causes of TBI-related deaths were firearm-related (40%), motor-vehicle-related (34%), and fall-related (10%). The leading causes of TBI-related deaths differed among age groups. Among youths aged 0-19 years, motor-vehicle-related TBIs were the leading cause; among persons aged 20-74 years, firearm-related TBIs were the leading cause; and among persons aged > or = 75 years, fall-related TBIs were the leading cause. Comparing rates in 1989 with rates in 1998, motor-vehicle-related causes declined by 22%; the majority of this decline occurred during the first 5 years of the period. During 1989-1998, firearm-related TBI-related deaths declined by 14%; approximately all of this decline occurred during the last 5 years of the period. In contrast, fall-related TBI-related death rates increased by 25% during the period. CONCLUSION: This analysis of mortality data identifies recent trends in TBI-related deaths occurring during 1989-1998. Fall-related TBI death rates have increased throughout the period. Firearm-related TBI death rates, which were increasing in the early 1990s, declined. Motor-vehicle-related TBI death rates, which were decreasing until the mid-1990s, have since demonstrated only a limited change. PUBLIC HEALTH ACTION: More current population-based epidemiologic studies of TBI are needed to assess recent trends of etiologic factors, provide additional guidance for public policy, and evaluate prevention strategies. Despite the decline in fatal TBI incidence, TBI morbidity and mortality remains a public health challenge. Public health, law enforcement, and transportation safety professionals can address these challenges by implementing effective interventions based on a thorough assessment of the factors that influence health related behaviors. PMID- 12529088 TI - Risk factors and secondary prevention in women with heart disease: the Heart and Estrogen/progestin Replacement Study. AB - BACKGROUND: Risk factors for coronary heart disease events have most commonly been evaluated in healthy men. OBJECTIVE: To assess risk factors, event rates, and use of secondary prevention treatments in women with preexisting coronary disease. DESIGN: A prospective cohort of clinical trial participants. SETTING: 20 U.S. clinical centers. PARTICIPANTS: 2763 postmenopausal women with known coronary disease in the Heart and Estrogen/progestin Replacement Study (HERS). MEASUREMENTS: Myocardial infarction or death from coronary heart disease. RESULTS: On multivariable analysis, the researchers found 11 risk factors: 6 noted by history (nonwhite ethnicity, lack of exercise, treated diabetes, angina, congestive heart failure, and more than one previous myocardial infarction) and 5 that were measured (blood pressure, low-density lipoprotein cholesterol level, high-density lipoprotein cholesterol level, lipoprotein(a) level, and creatinine clearance). The annual rate of coronary events was 1.3% (95% CI, 0.7% to 2.5%) in women with no risk factors and 8.7% (CI, 7.1% to 10.8%) in women with five or more risk factors (a sixfold increase). At entry into HERS, 83% of participants were receiving aspirin or other antiplatelet agents, 33% were receiving beta blockers, 18% were receiving angiotensin-converting enzyme inhibitors, and 53% were receiving lipid-lowering drugs. Women with more risk factors were less likely to be taking aspirin (P < 0.001) and lipid-lowering drugs (P = 0.006). CONCLUSIONS: Women with coronary disease are at high risk for myocardial infarction or death from coronary heart disease even in the absence of other risk factors, and their risk increases up to sixfold when many risk factors are present. Established drugs for secondary prevention, including aspirin, beta blockers, and lipid-lowering agents, are underused in these women, especially those at highest risk. PMID- 12529090 TI - Discrepancy between consensus recommendations and actual community use of adjuvant chemotherapy in women with breast cancer. AB - BACKGROUND: Although the efficacy of adjuvant chemotherapy in prolonging survival for women with breast cancer has been well documented, limited population-based information is available on the actual use of chemotherapy. OBJECTIVE: To examine the relationship between age and chemotherapy use. DESIGN: Cohort study. SETTING: New Mexico. PATIENTS: 5101 women 20 years of age or older receiving a diagnosis of stage I, stage II, or stage IIIA breast cancer from 1991 through 1997. MEASUREMENTS: Pattern of chemotherapy use by age; logistic regression analysis to generate the odds and probabilities of receiving chemotherapy; and sensitivity analysis to estimate potential effects of unmeasured confounders. RESULTS: Overall, 29% of women received chemotherapy. The rate of chemotherapy use for women with stage I, stage II, or stage IIIA breast cancer was 11%, 47%, and 68%, respectively. Across all tumor stages, the use of chemotherapy decreased substantially with increasing age (P < 0.001). Overall, 66% of women younger than 45 years of age received chemotherapy compared with 44% of women between 50 and 54 years of age, 31% of women between 55 and 59 years of age, and 18% of women between 60 and 64 years of age. The decreasing pattern of chemotherapy use with age continued after adjustment for prognostic factors and was relatively insensitive to changes in unmeasured factors. CONCLUSIONS: There is considerable discrepancy between the 1990 National Institutes of Health Consensus Conference recommendations for chemotherapy administration in women with breast cancer and the actual use of chemotherapy in the community. The decrease in use with age may relate to the decreasing efficacy of chemotherapy with age, as reported in clinical trials. Outcomes studies should address whether the recommendations are overly aggressive or whether practicing oncologists are too conservative in their use of chemotherapy. PMID- 12529091 TI - Pravastatin for secondary prevention of cardiovascular events in persons with mild chronic renal insufficiency. AB - BACKGROUND: Cardiovascular disease is a common cause of morbidity and death in persons with renal insufficiency. Although 3-hydroxy-3methylglutaryl coenzyme A reductase inhibitors (statins) are effective for secondary prevention of cardiovascular events in the general population, they have not been specifically studied in chronic renal insufficiency. OBJECTIVE: To determine whether pravastatin is effective and safe for secondary prevention of cardiovascular events in persons with chronic renal insufficiency. DESIGN: Post hoc subgroup analysis of a randomized, double-blind, placebo-controlled trial. SETTING: The Cholesterol and Recurrent Events (CARE) study, a randomized trial of pravastatin versus placebo in 4159 participants with previous myocardial infarction and total plasma cholesterol levels less than 6.21 mmol/L (<240 mg/dL). PARTICIPANTS: 1711 participants with chronic renal insufficiency defined by creatinine clearance less than or equal to 75 mL/min, using the Cockcroft-Gault equation. MEASUREMENTS: The primary outcome was death from coronary disease or symptomatic nonfatal myocardial infarction. RESULTS: After a median follow-up of 58.9 months, the incidence of the primary end point was lower in participants receiving pravastatin than in those receiving placebo (adjusted hazard ratio, 0.72 [95% CI, 0.55 to 0.95]; P = 0.02). Pravastatin was associated with lower adjusted hazard ratios for major coronary events (0.72 [CI, 0.59 to 0.88]; P = 0.001) and coronary revascularization (0.65 [CI, 0.50 to 0.83]; P = 0.001), but not total mortality (0.81 [CI, 0.61 to 1.08]; P = 0.14) or stroke (0.62 [CI, 0.39 to 1.00]; P = 0.051). Tests for interaction suggested that the observed benefit was independent of the presence and severity of renal insufficiency. Incidence of side effects was similar in persons receiving pravastatin and those receiving placebo. CONCLUSIONS: These data suggest that pravastatin is effective and appears safe for secondary prevention of cardiovascular events in persons with mild chronic renal insufficiency. Since statins may be underused in this setting, physicians should consider prescribing them for patients with chronic renal insufficiency and known coronary disease. PMID- 12529092 TI - Remission of chronic thrombotic thrombocytopenic purpura after treatment with cyclophosphamide and rituximab. AB - BACKGROUND: Thrombotic thrombocytopenic purpura (TTP) in adults is usually caused by autoantibody inhibitors of ADAMTS13. Treatment with plasma exchange is often effective but does not address the underlying autoimmune process. OBJECTIVE: To report the efficacy of intensive immunosuppressive therapy in refractory TTP. DESIGN: Case report. SETTING: University medical center. PATIENT: 42-year-old woman with chronic relapsing TTP. INTERVENTION: Immunosuppression therapy with rituximab and cyclophosphamide. MEASUREMENTS: ADAMTS13 activity and inhibitors and hematologic variables for TTP. RESULTS: For 19 months, the patient had relapsing thrombotic microangiopathy despite plasma exchange; splenectomy; and therapy with vincristine, prednisone, and cyclosporine. ADAMTS13 activity was low, and tests detected an IgG inhibitor that recognized the metalloprotease domain of recombinant ADAMTS13. After treatment with rituximab and cyclophosphamide, the disease remitted, ADAMTS13 levels normalized, and the inhibitor was undetectable. The patient has required no treatment for 13 months. CONCLUSION: Intensive immunosuppressive therapy can lead to sustained clinical remission in patients with refractory autoimmune TTP. PMID- 12529093 TI - Testing strategies in the initial management of patients with community-acquired pneumonia. AB - The initial management of patients suspected of having community-acquired pneumonia is challenging because of the broad range of clinical presentations, potential life-threatening nature of the illness, and associated high costs of care. The initial testing strategies should accurately establish a diagnosis and prognosis in order to determine the optimal treatment strategy. The diagnosis is important in determining the need for antibiotic therapy, and the prognosis is important in determining the site of care. This paper reviews the test characteristics of the history, physical examination, and laboratory findings, individually and in combination, in diagnosing community-acquired pneumonia and predicting short-term risk for death from the infection. In addition, we consider the implications of these test characteristics from the perspective of decision thresholds. The history and physical examination cannot provide a high level of certainty in the diagnosis of community-acquired pneumonia, but the absence of vital sign abnormalities substantially reduces the probability of the infection. Chest radiography is considered the gold standard for pneumonia diagnosis; however, we do not know its sensitivity and specificity, and we have limited data on the costs of false-positive and false-negative results. In the absence of empirical evidence, the decision to order a chest radiograph needs to rely on expert opinion in seeking strategies to optimize the balance between harms and benefits. Once community-acquired pneumonia is diagnosed, a combination of history, physical examination, and laboratory items can help estimate the short term risk for death and, along with the patient's psychosocial characteristics, determine the appropriate site of treatment. PMID- 12529094 TI - Update in women's health. PMID- 12529095 TI - Congenital thrombophilic states associated with venous thrombosis: a qualitative overview and proposed classification system. AB - Congenital causes of venous thrombosis have gained increasing prominence with the description of the factor V Leiden mutation and the prothrombin gene mutation. More recently, the description of the association between increased levels of coagulation factors and venous thrombosis and the finding that patients with thrombophilia can harbor more than one prothrombotic state have further increased the clinical relevance of the congenital thrombophilic states. In this qualitative review, we summarize current knowledge of the congenital prothrombotic states and propose a simple classification system that divides the states into two broad groups: those associated with reduced levels of the inhibitors of the coagulation cascade and those associated with increased levels or function of the coagulation factors. The first group is less common than the second, but it is associated with a much higher risk for venous thrombosis. This review provides clinicians with an evidence-based, practical guide to the congenital prothrombotic states. PMID- 12529096 TI - Linezolid: the first oxazolidinone antimicrobial. AB - Linezolid is the first of a new class of antimicrobial agents, the oxazolidinones, to be approved for clinical use in the United States and elsewhere. The drug is a totally synthetic compound, which lessens the likelihood of naturally occurring resistance mechanisms. It has excellent activity against virtually all important gram-positive pathogens, including methicillin-resistant staphylococci, penicillin-resistant pneumococci, macrolide-resistant streptococci, and vancomycin-resistant enterococci. Development of resistance to the compound has been infrequent thus far. Linezolid is 100% bioavailable, so it can be given in equal doses orally or parenterally. Its elimination half-life allows dosing twice per day, and alteration of drug dosage is not required in patients with impaired renal or hepatic function. Linezolid has approved indications for skin and soft tissue infections; lower respiratory tract infections; and vancomycin-resistant Enterococcus faecium infections, including cases with concurrent bacteremia. The drug has an acceptable profile of adverse events, but reversible myelosuppression has occurred in patients receiving high doses for more than 2 weeks. PMID- 12529097 TI - Denver Health: a model for the integration of a public hospital and community health centers. AB - Two major pillars of the United States' safety net system are urban public hospitals and community health centers. Their common mission is to care for the uninsured and other vulnerable populations. However, in most communities these important components of the safety net remain organizationally and functionally separate, which inhibits the continuum of care and creates substantial inefficiencies. Denver Health is a long-standing vertically and horizontally integrated system for vulnerable populations. The integration benefits the patient and the system and serves as a model for the U.S. safety net. This paper outlines the benefits of integration to the patient, provider, and health system, using data from the National Association of Public Hospitals and Health Systems, the Bureau of Primary Health Care, and Denver Health. PMID- 12529098 TI - Secondary prevention of coronary heart disease in women: a call to action. PMID- 12529099 TI - Thrombotic thrombocytopenic purpura: from the bench to the bedside, but not yet to the community. PMID- 12529100 TI - A memorable patient. PMID- 12529101 TI - Healthy ranges for alanine aminotransferase levels. PMID- 12529102 TI - Healthy ranges for alanine aminotransferase levels. PMID- 12529104 TI - Adrenalectomy for primary aldosteronism. PMID- 12529106 TI - Did this drug cause my patient's hepatitis? PMID- 12529108 TI - Amelioration of microvascular angina with arginine supplementation. PMID- 12529109 TI - Summaries for patients. Linezolid. PMID- 12529110 TI - Summaries for patients. Congenital causes of venous thrombosis--a classification system. PMID- 12529111 TI - Summaries for patients. Treatment of refractory thrombotic thrombocytopenic purpura. PMID- 12529112 TI - Summaries for patients. Treating cholesterol in patients with mild kidney disease. PMID- 12529113 TI - Summaries for patients. Chemotherapy in women with breast cancer. PMID- 12529114 TI - Summaries for patients. Risk factors and treatment in women with coronary artery disease. PMID- 12529115 TI - Hydrophobic polymer-supported catalyst for organic reactions in water: acid catalyzed hydrolysis of thioesters and transprotection of thiols. AB - [reaction: see text] A hydrophobic polystyrene-supported sulfonic acid was found to be effective for hydrolysis of thioesters in pure water. It was revealed that the catalyst was much superior to other Bronsted acid catalysts. Transprotection of thiols from thioesters to thioethers has been successfully performed in water using this catalytic system. PMID- 12529116 TI - ROMP-generated oligomeric sulfonyl chlorides as versatile soluble scavenging agents. AB - [reaction: see text] A new method for homogeneous nucleophilic scavenging employing oligomeric sulfonyl chloride (OSC) reagents is described. The method utilizes OSC to rapidly scavenge a variety of amines that are present in excess. The OSC reagents are generated from ROM polymerization of 2-chlorosulfonyl-5 norbornene utilizing the second generation Grubbs catalyst to produce oligomers of varying size as stable, free-flowing powders. Following the scavenging event, these oligomers are precipitated with ethyl acetate leaving products in excellent yield and purity. PMID- 12529117 TI - Novel antibiotics: macrocyclic peptides designed to trap Holliday junctions. AB - [reaction: see text] Described are the syntheses of eight macrocyclic peptides designed to trap Holliday junctions in bacteria, thereby inhibiting bacterial growth. These macrocycles were designed from linear dimerized hexapeptides that bind to the C-2 symmetrical Holliday junction. They were synthesized from three monomers using a combinatorial-like strategy that permits elucidation of the monomer role in accumulation of Holliday junctions and antibiotic activity. These macrocycles are an important step in designing and synthesizing a new class of antibiotics. PMID- 12529118 TI - Zinc and copper complexes of prodigiosin: implications for copper-mediated double strand DNA cleavage. AB - [reaction: see text] Zinc(II) and copper(II) complexes of prodigiosin (1) have been characterized. All N-atoms of 1 bind Cu(II) to generate 5: the complex exhibits regiospecific oxidation of the C-pyrrole. In contrast, coordination by Zn(II) to 1 produces Zn(1)(2) (8), a 4-coordinate tetrahedral complex. The influence of these binding geometries on Cu-mediated double-strand (ds) DNA cleavage by 1 is discussed. PMID- 12529119 TI - Novel orthogonal strategy toward solid-phase synthesis of 1,3,5-substituted triazines. AB - [reaction: see text] To improve upon the previous orthogonal method for synthesis of a triazine library, an alternative strategy has been developed via oxidation activation of the thioether to the sulfone. Through a comparison between these two methods, the sulfone strategy was demonstrated as an enhanced method in the generation of highly pure triazine library compounds. PMID- 12529120 TI - Synthesis of long-chain fatty acid enol esters isolated from an environmental DNA clone. AB - [structure: see text] Long-chain fatty acid enol ester 1 is the major metabolite of a new family of small molecules isolated from the heterologous expression of environmentally derived DNA. A versatile synthesis of 1, in which an aromatic acetaldehyde is O-acylated with a long-chain acyl chloride allowed for the rapid construction of both the isolated product (1) and a number of structural analogues (including 8, 17, and 18). PMID- 12529121 TI - An efficient and practical N-methylation of amino acid derivatives. AB - [reaction: see text] An efficient and practical N-methylation of amino acid derivatives with dimethyl sulfate in the presence of sodium hydride and a catalytic amount of water is described. Reaction of water with sodium hydride generated highly reactive dry sodium hydroxide, which led to much faster reaction rates than powdered sodium hydroxide itself. PMID- 12529122 TI - Novel phosphine-catalyzed zipper cyclization of aliphatic diyne-dione and yne dione systems. AB - [reaction: see text] An unexpected tri-n-butylphosphine-catalyzed zipper cyclization of diyne[bond]diones (1a-d) or yne-diones (1e and 1f) is described. Bicyclic ketones (2a, 2b, 2c, 2e, and 2f) with five- or six-membered rings fused to the five-membered ring were obtained from both aliphatic diyne-diones (1a-c) and yne-diones (1e and 1f) having tetra- or pentamethylene spacers. The bicyclic products (2) were produced with high diastereoselectivity. PMID- 12529123 TI - Copper- and palladium-catalyzed intramolecular aryl guanidinylation: an efficient method for the synthesis of 2-aminobenzimidazoles. AB - [reaction: see text] The formation of 2-aminobenzimidazoles via intramolecular C[bond]N formation between an aryl halide and a guanidine moiety can be achieved using either copper or palladium catalysis. Inexpensive copper salts such as CuI are generally superior to the use of palladium catalysts. Regioselective cyclizations, where R(3) = H, can be achieved in high yield under CuI/1,10 phenanthroline-catalyzed conditions, whereas palladium catalysis results in the formation of regioisomeric products. PMID- 12529124 TI - First synthesis of the A/B ring of ouabain. AB - [reaction: see text] The synthesis of the fully fuctionalized A/B ring of ouabain has been accomplished efficiently from commercially available starting materials. A key Robinson annulation allows for the building of the desired carbon framework in one high-yielding step. Directed epoxidation followed by selective epoxide opening furnished the final tetraol with the desired all-cis stereochemistry. PMID- 12529125 TI - Synthesis and characterization of novel bifunctional hemithioindigo chromophores. AB - [structure: see text] General methods for the synthesis of novel bifunctional hemithioindigo (HT) compounds, e.g., omega-amino acid derivatives, are presented. The photochromic properties of the photoswitches have been characterized by UV vis and (1)H NMR spectroscopy. PMID- 12529126 TI - Copper(I)-promoted dechlorinative Surzur-Tanner rearrangement of 2,2,2 trichloroethyl carboxylates. AB - [reaction: see text] 2,2,2-Trichloroethyl carboxylates undergo highly efficient dechlorinative Surzur-Tanner rearrangement with 2 equiv of a 1:1 molar mixture of CuCl and bpy in boiling DCE to give 1-chloroethenyl carboxylates in which copper appears to play an important role, probably by coordinating the initial radical or as a Lewis acid catalyst. PMID- 12529127 TI - Calix[4]arene-porphyrin conjugates as versatile molecular receptors for anions. AB - [structure: see text] Appending tetraphenylporphyrin units to the calix[4]arene skeleton via the ureido function leads to novel anion receptors designed for anion and/or cation detection by UV-vis spectroscopy. Calixarenes in the cone or 1,3-alternate conformations bearing two ureido moieties on the upper rim represent well-preorganized cavities where the anion can be held by synchronous hydrogen bonding interaction with the NH groups. PMID- 12529128 TI - Triethyl- (or trimethyl-)silyl triflate-catalyzed reductive cleavage of triphenylmethyl (trityl) ethers with triethylsilane. AB - [reaction: see text] A triphenylmethyl (trityl) ether was reductively and instantaneously cleaved by triethylsilane in the presence of a catalytic amount of TES- (or TMS)-triflate. The reaction conditions are mild enough to achieve reduction in the presence of a variety of acid-sensitive functional groups. Upon a mild acidic treatment of the crude product, the corresponding alcohol is obtained in high yield. PMID- 12529129 TI - Enantioselective cyclization/hydrosilylation of 1,6-enynes catalyzed by a cationic rhodium bis(phosphine) complex. AB - [reaction: see text] Reaction of 4,4-dicarbomethoxy-1-octene-6-yne (1) with triethylsilane and a catalytic 1:1 mixture of [Rh(COD)(2)](+) SbF(6)(-) and (R) BIPHEMP (5 mol %) at 70 degrees C for 90 min gave (Z)-1,1-dicarbomethoxy-3-(1 triethylsilyl)ethylidene-4-methylcyclopentane (2) in 81% isolated yield with 98% de and 92% ee. PMID- 12529130 TI - Diastereoselective synthesis of a conformationally restricted dinucleotide with predefined alpha and beta torsional angles for the construction of alpha,beta constrained nucleic acids (alpha,beta-CNA). AB - [reaction: see text] The synthesis of a diastereopure 1,3,2-dioxaphosphorinane linkage in which two, alpha and beta, out of six torsion angles of the natural phosphodiester backbone are constrained with predefined values of ca. +60 degrees (g(+)) and 180 degrees (t), respectively, is described. The stable and unstrained six-membered cyclic phosphotriester structure represents the smallest possible ring allowing the conformational locking of the alpha torsion angle at a significant positive value that is typical of many bulged regions of nucleic acids. PMID- 12529131 TI - Formation of the C ring in the lanosterol biosynthesis from squalene. AB - [reaction: see text] Ab initio calculations were performed on a cyclohexane derivative to elucidate the mechanism of the formation of the five-membered C ring in the biosynthesis of lanosterol from squalene. A conformational analysis of the side chain containing the double bond indicated that the conformer that should give rise to the cyclized C ring is not a minimum on the potential surface. Consequently, it is suggested that it is very likely that C-ring formation occurs in concert with formation of the A and B rings. PMID- 12529132 TI - (-)-Axinyssene: a novel cytotoxic diterpene from a Japanese marine sponge Axinyssa sp. AB - [structure: see text] A novel diterpene, (-)-axinyssene, was isolated from the Japanese marine sponge Axinyssa sp. The structure of (-)-axinyssene was determined on the basis of spectroscopic and synthetic evidence to be 1-methyl-4 [(4E)-5',9'-dimethyl-1'-methylene-4',8'-decadienyl]-(4S)-cyclohexene. (-)- and (+)-axinyssene showed mild cytotoxicity against acute promyelocytic leukemia, HL 60 cells. PMID- 12529133 TI - Practical asymmetric enzymatic reduction through discovery of a dehydrogenase compatible biphasic reaction media. AB - [reaction: see text] An enzyme-compatible biphasic reaction media for the asymmetric biocatalytic reduction of ketones with in situ cofactor regeneration has been developed. In this biphasic reaction media, which is advantageous for reactions at higher substrate concentrations, both enzymes (alcohol dehydrogenase and FDH from Candida boidinii) remain stable. The reductions with poorly water soluble ketones were carried out at substrate concentrations of 10-200 mM, and the optically active (S)-alcohols were formed with moderate to good conversions and with up to >99% ee. PMID- 12529134 TI - Contrasting diastereofacial selectivity associated with N-phenyltriazolinedione cycloadditions to oxaspirocycloheptatrienes. AB - [reaction: see text] The cycloaddition of N-phenyltriazolinedione to a pair of spirocyclic cycloheptatrienes featuring a tetrahydrofuran or tetrahydropyran ring is shown to proceed with opposite pi-facial stereoselectivity. In addition, the furan undergoes direct [4+2] cycloaddition to the cycloheptatriene whereas the pyran product is a diazetidine. PMID- 12529135 TI - Toward amide-linked RNA mimics: total synthesis of 3'-C branched uridine azido acid via an ene-iodolactonization approach. AB - [reaction: see text] A novel synthesis of 3'-C branched uridine azido acid has been accomplished by a sequence of stereoselective ene and iodolactonization reactions. Compared to traditional routes that start from carbohydrates, the present methodology is more flexible and can be further optimized by incorporation of novel future developments of synthetic chemistry. Because the key chemistry does not involve the 3'-C substituent, our route is a general approach to 3'-C alkyl nucleoside analogues. PMID- 12529136 TI - Microencapsulation of osmium tetroxide in polyurea. AB - [reaction: see text] Osmium tetroxide has been microencapsulated in a polyurea matrix using an in situ interfacial polymerization approach. These microcapsules have been effectively used as recoverable and reusable catalysts in the dihydroxylation of olefins PMID- 12529137 TI - The reaction of cyclic carbinol amides with triflic anhydride as a method to prepare alpha-trifluoromethyl-sulfonamido furans. AB - [reaction: see text] A novel synthesis of alpha-trifluoromethyl-sulfonamido furans via the reaction of cyclic carbinol amides with triflic anhydride has been developed. The reaction proceeds under very mild conditions with a wide set of representative lactams to provide the alpha-trifluoromethyl-sulfonamido substituted furan in high yield. Rapid access to a 5-substituted indoline can be achieved by thermolysis of the N-but-3-enyl-substituted sulfonamido furan. PMID- 12529138 TI - Uncatalyzed Friedel-Crafts alkylation of aromatic compounds through reactive benzyl cations generated from N-sulfamoylcarbamates. AB - [reaction: see text] A new method for the generation of highly reactive benzyl cations by thermal decomposition of aryl-benzyl-sulfamoylcarbamates, obtained in a one-pot reaction from chlorosulfonyl isocyanate, is described. The generated cations alkylate aromatic compounds efficiently in the absence of catalysts. PMID- 12529139 TI - A concise formal total synthesis of TMC-95A/B proteasome inhibitors. AB - [reaction: see text] A formal total synthesis of proteasome inhibitors TMC-95A/B is described. The synthesis features a stereoselective modified Julia olefination and a diastereoselective dihydroxylation to construct the highly oxidized tryptophan residue. PMID- 12529140 TI - Microwave-promoted Pd-catalyzed cyanation of aryl triflates: a fast and versatile access to 3-cyano-3-desoxy-10-ketomorphinans. AB - [reaction: see text] A methodology for the microwave-promoted conversion of triflates to the corresponding nitriles by using 8% equiv of Pd(Ph(3)P)(4) as the catalyst and 2.0 equiv of Zn(CN)(2) as the cyanide source has been developed. This method is highly efficient for the preparation of 3-cyano-3-desoxy-10 ketomorphinans. The reaction was generally completed in 15 min at 200 degrees C and produced products in greater than 86% yields. PMID- 12529141 TI - Asymmetric hydrogenation of itaconic acid and enol acetate derivatives with the Rh-TangPhos catalyst. AB - [reaction: see text] The Rh-TangPhos catalyst has been used for asymmetric hydrogenation of itaconic acid and enol acetate derivatives. A variety of chiral 2-substituted succinic acids and chiral acetates have been obtained in excellent ee values (up to 99% ee). PMID- 12529142 TI - A novel and convenient chemoselective deprotection method for both silyl and acetyl groups on acidic hydroxyl groups such as phenol and carboxylic acid by using a nitrogen organic base, 1,1,3,3-tetramethylguanidine. AB - [reaction: see text] 1,1,3,3-Tetramethylguanidine (TMG)(1), a nitrogen organic base, is a convenient and useful reagent for chemoselective deprotection of both silyl and acetyl groups on acidic hydroxyl groups such as phenol and carboxylic acid without affecting aliphatic silyl and acetyl groups. The chemoselectivity is dependent on the acidity of the hydroxyl group. PMID- 12529143 TI - Synthesis and derivatization of ethynyl alpha,alpha-dibromomethyl ketones: formation of highly functionalized vinyl triflates. AB - [reaction: see text] We describe the synthesis of alpha,alpha-dibromomethyl ynones (8) and their subsequent derivatization to vinyl acetates (10). These vinyl acetates feature a 1,1-dibromo-olefin moiety, which is readily exploited in palladium-catalyzed Sonogashira, Stille, and Suzuki cross-coupling reactions with alkynes, stannanes, and boronic acids, respectively. A novel one-pot process then directly converts the resulting vinyl acetates 11-13 to the vinyl triflate derivatives 14a-j. PMID- 12529144 TI - Palladium-catalyzed asymmetric synthesis of axially chiral (allenylmethyl)silanes and chirality transfer to stereogenic carbon centers in S(E)' reactions. AB - [reaction: see text] Novel stereoselective reactions of 4-substituted-1 trimethylsilyl-2,3-butadienes ((allenylmethyl)silanes) were developed. The axially chiral (allenylmethyl)silanes were prepared from (3-bromopenta-2,4 dienyl)trimethylsilane by a Pd-catalyzed asymmetric reaction with soft nucleophiles with up to 88% enantioselectivity. The (allenylmethyl)silanes reacted with acetals in the presence of a TiCl(4) promoter to give 1,3-diene derivatives via an S(E)' pathway. The 1,3-dienyl products have (E)-geometry exclusively and up to 88%( )()chirality transfer from the axially chiral allenes to the centrally chiral 1,3-dienes was observed in the S(E)' reaction. PMID- 12529145 TI - Zirconocene-mediated route to enantiopure 9-oxabicyclononanes functionalized on both carbon bridges. AB - [reaction: see text] A zirconocene-mediated ring contraction of 4 vinylfuranosides generated either from d-arabinose or d-glucose is followed by sequential oxidation to the ketone and alkynyl Grignard addition. The resulting cis-cyclobutanediols are subjected in turn to thermal rearrangement and intramolecular oxymercuration-demercuration. The regiochemistry of the final ring closure is controlled by the nature of R. PMID- 12529146 TI - gem-Silylborylation of an sp carbon: novel synthesis of 1-boryl-1-silylallenes. AB - [reaction: see text] Novel synthesis of 1-boryl-1-silylallenes involving gem silylborylation of 3-chloro- or 3-alkoxyalkyn-1-yllithiums with (dimethylphenylsilyl)(pinacolato)borane has been established. The reaction proceeds via 1,2-migration of the silyl group from the negatively charged boron atom of an intermediary borate complex to the terminal acetylenic carbon and is accelerated by the addition of chlorotrimethylsilane in the case that methanesulfonyloxy is employed as a leaving group. Furthermore, axially enantioenriched products could be prepared from mesylates of optically active propargylic alcohols. PMID- 12529147 TI - SmI(2)-promoted radical addition of nitrones to alpha,beta-unsaturated amides and esters: synthesis of gamma-amino acids via a nitrogen equivalent to the ketyl radical. AB - [reaction: see text] Alkyl nitrones undergo radical addition reactions to a series of alpha,beta-unsaturated amides and esters when subjected to samarium diiodide via a nitrogen equivalent to a ketyl radical anion. This reaction conveniently provides access to a variety of functionalized gamma-amino acids. The methodology was extended to the asymmetric synthesis of 4-substituted gamma amino acids, via the nitrone radical addition reaction to acrylates/amides possessing a chiral auxiliary. PMID- 12529149 TI - Force field dependence of NMR-Based, restrained molecular dynamics DNA structure calculations including an analysis of the influence of residual dipolar coupling restraints. AB - Restrained molecular dynamics is widely used to calculate DNA structures from NMR data. Here, results of an in silico experiment show that the force field can be significant compared to the NMR restraints in driving the final structures to converge. Specifically, we observed that i) the influence of the force field leads to artificially tight convergence within final families of structures and ii) the precision and character of resulting structures depend on the choice of force field used in the calculations. A canonical B-DNA model was used as a target structure. Distances, dihedral angles, and simulated residual dipolar couplings were measured in the target structure and used as restraints. X-PLOR and Discover, which use force fields developed for CHARMM and AMBER programs, respectively, were tested and found to produce different final structures despite the use of identical distance and dihedral restraints. Incorporation of residual dipolar coupling restraints in X-PLOR improves convergence with the target structure and between families of structures indicating that the force field dependence can potentially be overcome if residual dipolar coupling restraints are employed. PMID- 12529150 TI - Interaction of cyclic cytosine-, guanine-, thymine-, uracil- and mixed guanine cytosine base tetrads with K+, Na+ and Li+ ions -- a density functional study. AB - We have carried out B3LYP hybrid density functional studies of complexes formed by cyclic cytosine-, guanine-, thymine-, uracil- and mixed guanine cytosine tetrads with Li+, Na+ and K+ ions to determine their structures and interaction energies. The conformations studied have been restricted to a hydrogen bond pattern closely related to the tetrads observed in experimental nucleic acid structures. A comparison of the alkali metal ion/tetrad complexes with the tetrads without cations indicates that alkali metal ions modulate the tetrad structures significantly and that even the hydrogen bond pattern may change. Guanine-tetrad cation complexes show the strongest interaction energy compared to other tetrads that occur less frequently in experimental structures. The most stable G-tetrad/metal ion structure adopts a nearly planar geometry that is especially suitable for tetraplex formation, which requires approximately parallel tetrad planes. In the cytosine-tetrad there is a very large central cavity suitable for cation recognition, but the complexes adopt a non-planar structure unsuitable for stacking, except possibly for ions with very large radii. Uracil and thymine tetrads show a significant different characteristics which may contribute to the differences between DNA and RNA PMID- 12529151 TI - Thermodynamics of DNA containing very stable chemically modified base pairs. AB - DNA chemical modifications caused by the binding of some antitumor drugs give rise to a very strong local stabilization of the double helix. These sites melt at a temperature that is well above the melting temperatures of ordinary AT and GC base pairs. In this work we have examined the melting behavior of DNA containing very stable sites. Analytical expressions were derived and used to evaluate the thermodynamic properties of homopolymer DNA with several different distributions of stable sites. The results were extended to DNA with a heterogeneous sequence of AT and GC base pairs. The results were compared to the melting properties of DNA with ordinary covalent interstrand cross-links. It was found that, as with an ordinary interstrand cross-link, a single strongly stabilized site makes a DNA's melting temperature (T(m)) independent of strand concentration. However in contrast to a DNA with an interstrand cross-link, a strongly stabilized site makes the DNA's T(m) independent of DNA length and equal to T(infinity), the melting temperature of an infinite length DNA with the same GC-content and without a stabilized site. Moreover, at a temperature where more than 80% of base pairs are melted, the number of ordinary (non-modified) helical base pairs (n) is independent of both the DNA length and the location of the stabilized sites. For this condition, n(T) = (2 omega-a)S/(1-S) and S = exp[DeltaS(T(infinity)-T)/(RT)] where omega is the number of strongly stabilized sites in the DNA chain, a is the number of DNA ends that contain a stabilized site, and DeltaS, T, and R are the base pair entropy change, the temperature, and the universal gas constant per mole. The above expression is valid for a temperature interval that corresponds to n<0.2N for omega=1, and n<0.1N for omega>1, where N is the number of ordinary base pairs in the DNA chain. PMID- 12529152 TI - Melting of cross-linked DNA v. cross-linking effect caused by local stabilization of the double helix. AB - DNA interstrand cross-links are usually formed due to bidentate covalent or coordination binding of a cross-linking agent to nucleotides of different strands. However interstrand linkages can be also caused by any type of chemical modification that gives rise to a strong local stabilization of the double helix. These stabilized sites conserve their helical structure and prevent local and total strand separation at temperatures above the melting of ordinary AT and GC base pairs. This local stabilization makes DNA melting fully reversible and independent of strand concentration like ordinary covalent interstrand cross links. The stabilization can be caused by all the types of chemical modifications (interstrand cross-links, intrastrand cross-links or monofunctional adducts) if they give rise to a strong enough local stabilization of the double helix. Our calculation demonstrates that an increase in stability by 25 to 30 kcal in the free energy of a single base pair of the double helix is sufficient for this "cross-linking effect" (i.e. conserving the helicity of this base pair and preventing strand separation after melting of ordinary base pairs). For the situation where there is more then one stabilized site in a DNA duplex (e.g., 1 stabilized site per 1000 bp), a lower stabilization per site is sufficient for the "cross-linking effect" (18 - 20 kcal). A substantial increase in DNA stability was found in various experimental studies for some metal-based anti tumor compounds. These compounds may give rise to the effect described above. If ligand induced stabilization is distributed among several neighboring base pairs, a much lower minimum increase per stabilized base pair is sufficient to produce the cross-linking effect (1 bp- 24.4 kcal; 5 bp- 5.3 kcal; 10 bp- 2.9 kcal, 25 bp 1.4 kcal; 50 bp- 1.0 kcal). The relatively weak non-covalent binding of histones or protamines that cover long regions of DNA (20- 40 bp) can also cause this effect if the salt concentration of the solution is sufficiently low to cause strong local stabilization of the double helix. Stretches of GC pairs more than 25 bp in length inserted into poly(AT) DNA also exhibit properties of stabilizing interstrand cross-links. PMID- 12529153 TI - The conformer substates of nonoriented B-type DNA in double helical poly(dG-dC). AB - A nonoriented hydrated film of poly(dG-dC) with ?20 water molecules per nucleotide (called B* by Loprete and Hartman (Biochem. 32, 4077-4082 (1993)) was studied by Fourier transform infrared (FT-IR) spectroscopy either as equilibrated sample between 290 and 270 K or, after quenching into the glassy state, as nonequilibrated film isothermally at 200 and 220 K. IR spectral changes on isothermal relaxation at 200 and 220 K, caused by interconversion of two conformer substates, are revealed by difference spectra. Comparison with difference curves obtained in the same manner from two classical B-DNA forms, namely the d(CGCGAATTCGCG)(2) dodecamer and polymeric NaDNA from salmon testes, revealed that the spectral changes on B(I)-to-B(II) interconversion in the classical B-DNA forms are very similar to those in the B*-form, and that the spectroscopic differences between the B(I) and B(II) features from classical B DNA and those from the modified B*-form are minor. Nonexponential kinetics of the B(I)-->B(II) transition in the B*-form of poly(dG-dC) at 200 K showed that the structural relaxation time is about three times of that in the classical B-DNA forms (approximately equal to 30 versus approximately equal to 10 min at 200 K). The unexpected reversal of conformer substates interconversion (that is B(II)- >B(I) transition on cooling from 290 K and B(I)-->B(II) transition on isothermal relaxation at 200 K) observed for classical B-DNA occurs also in the modified B* form. We therefore conclude that restructuring of hydration shells rules the low temperature dynamics of the B*-form via its two conformer substates in the same manner reported for classical B-DNA by Pichler et al. (J. Phys. Chem. B 106, 3263 3274 (2002)). PMID- 12529154 TI - Thallium-DNA complexes in aqueous solution. Major or minor groove binding. AB - Thallium (Tl) binds to the major and minor grooves of B-DNA in the solid state (Howerton et al., Biochemistry 40, 10023-10031, 2001). The aim of this study was to examine the binding of Tl(I) cation with calf-thymus DNA in aqueous solution at physiological pH, using constant concentration of DNA (12.5 mM) and various concentrations of metal ions (0.5 to 20 mM). UV-visible and FTIR spectroscopic methods were used to determine the cation binding site, the binding constant and DNA structural variations in aqueous solution. Direct Tl bindings to guanine and thymine were evident by major spectral changes of DNA bases with overall binding constant of K = 1.40 x 10(4) M(-1) and little perturbations of the backbone phosphate group. Both major and minor groove bindings were observed with no alteration of the B-DNA conformation. At low metal concentration (0.5 mM), the number of cations bound were 10 per 1000 nucleotides, while at higher cation concentration (10 mM), this increased to 30 cations per 1000 nucleotides. PMID- 12529155 TI - Molecular dynamics simulations of the unfolding mechanism of the catalytic domain from Aspergillus awamori var. X100 glucoamylase. AB - In this study, 200 ps molecular dynamics simulations were conducted to investigate the unfolding mechanism of the catalytic domain of glucoamylase from Aspergillus awamori var. X100. The unfolding of this domain was suggested to follow a putative hierarchical manner, in which the heavily O-glycosylated belt region from residues T440 to A471 acted as the initiation site, followed by the alpha-helix secondary structure destruction, and then the collapse of the catalytic center pocket. The O-glycosylated belt region surrounded the surface of the catalytic domain in its native state at low temperature, whereas it was extended and is more suitable to be classified as part of the subsequent linker domain at high temperatures due to its high flexibility. The inner set helices of the (alpha/alpha)(6)-barrel seemed to exhibit higher helical content than the outer set ones at all temperatures examined. The distances between the C(alpha) of the three Cys residue pairs fluctuated rapidly at higher temperatures, indicating that these disulfide bonds have little effect on the structural stabilization. The melting temperature, at which the residual total helicity of the catalytic domain is 50%, is much lower than the critical temperature, at which the catalytic center pocket has lost its structural integrity. PMID- 12529156 TI - Structural modification and aggregation of mucin by chromium(III) complexes. AB - Metal ions binding to proteins regulate the functions of proteins and may also lead to structural changes. In this communication we demonstrate the interaction and subsequent conformational changes induced in pig gastric mucin (PGM) upon binding to certain chromium(III) complexes like, [Cr(salen)(H(2)O)(2)](ClO(4)) (1), [Cr(en)(3)]Cl(3) (2) and [Cr(EDTA)(H(2)O)]Na (3) which vary in charge and ionic character. Complexes 1 and 3 have been shown to interact coordinately with PGM whereas complex 2 binds through electrostatic interaction and hydrogen bonding. Steady state fluorescence experiment reveals that at lower concentration of complex 2 there is partial quenching of the tyrosine emission, whereas at higher concentration of the complex the emission intensity is enhanced. On the other hand with complexes 1 and 3 a decrease in fluorescence intensity was observed. PGM viscosity was found to decrease in the presence of complex 1 and 3 due to the formation of flexible fibres through coordinate interaction. Complex 2 was found to facilitate metal induced intertangling of PGM fibres which tends to stabilize the interaction and leads to sol-gel transition with subsequent increase in viscosity. A significant change in CD spectrum of PGM was observed in the presence of complex 2, where random coil spectrum became typical of a alpha helical structure with 80% alpha helix content. In the case of complexes 1 and 3 only minor changes in the amplitude of the spectrum were observed. Histochemical analysis supports the contention that complex 2 favors the oligomerisation of PGM and leads to the formation of aggregated mass of macromolecules. PMID- 12529157 TI - Exploring the interaction energies for the binding of hydroxydiphenyl ethers to enoyl-acyl carrier protein reductases. AB - It is now well established that the potent anti-microbial compound, triclosan, interrupts the type II fatty acid synthesis by inhibiting the enzyme enoyl-ACP reductase in a number of organisms. Existence of a high degree of similarity between the recently discovered enoyl-ACP reductase from P. falciparum and B. napus enzyme permitted building of a satisfactory model for the former enzyme that explained some of the key aspects of the enzyme such as its specificity for binding to the cofactor and the inhibitor. We now report the interaction energies between triclosan and other hydroxydiphenyl ethers with the enzymes from B. napus, E. coli and P. falciparum. Examination of the triclosan-enzyme interactions revealed that subtle differences exist in the ligand binding sites of the enzymes from different sources i.e., B. napus, E. coli and P. falciparum. A comparison of their binding propensities thus determined should aid in the design of effective inhibitors for the respective enzymes. PMID- 12529158 TI - Secondary conformations and temperature effect on structural transformation of amyloid beta (1-28), (1-40) and (1-42) peptides. AB - Secondary structure of three amyloid b-peptides [A beta(1-28), A beta(1-40) and A beta(1-42)] in the solid state was respectively determined by Fourier transform infrared (FT-IR) microspectroscopy. Their thermal-dependent structural transformation were also investigated by FT-IR microspectroscopy equipped with a thermal analyzer. The present result demonstrates that the solid-state A beta(1 28), A beta(1-40) and A beta(1-42) peptides showed a significant IR spectral difference in the amide I and II bands. The secondary conformation of A beta(1 28) peptide was the combination of major beta-sheet and minor alpha-helix with little random coil structures, but A beta(1-40) peptide showed the co-existence of major beta-sheet and minor random coil with little alpha-helix structures. A beta(1-42) peptide mainly consisted of the predominant b-sheet structure. Although the intact A beta(1-28), A beta(1-40) or A beta(1-42) peptide exhibits a different secondary structure, a similar beta-conformation may form after thermal treatment. A thermal-dependent transition was found for solid A beta(1-28) and A beta(1-40) peptides near 40 degrees C and 45 degrees C, respectively. There was no transition temperature for solid A beta(1-42) peptide, however, due to only a very little level of alpha-helix and random coil structure containing in the solid A beta(1-42) peptide. The thermal denaturation plays an important role in the structural transformation from alpha-helix/random coil to beta-sheet. PMID- 12529159 TI - Structure and polymorphism of the principal neutralization site of Thailand HIV-1 isolate. AB - Refining the geometric parameters for the ensemble of conformers, derived earlier in terms of NMR-spectroscopy data for the immunogenic tip of Thailand HIV-1 isolate, was carried out by quantum chemical methods. As a result, (i) the energy characteristics of initial structures were significantly improved, (ii) their relative locations on the scale of formation heats were determined, and (iii) the energy barriers between conformers under study were computed. On the basis of all data obtained, the high resolution 3D structure model, describing the set of stable conformers and containing the biologically active conformation, was proposed for neutralizing epitope of Thailand HIV-1 isolate. The following major conclusions were made based on the analysis of simulated conformations: i) the Gly-Pro-Gly-Gln-Val-Phe stretch forming the immunogenic crown of Thailand HIV-1 isolate exhibits the properties characteristic for metastable oligopeptide that constitutes in solution the dominant structure with other conformations admissible; (ii) three structures out of five NMR-based starting models form the cluster of conformers which adequately describes general conformational features of this functionally important site of gp120; (iii) two structures residing in this cluster are found to be well-ground for implementing the function of immunoreactive conformation of the stretch of interest; (iv) in spite of this observation, the "global" structure which gives rise to inverse gamma-turn in the central Gly-Pro-Gly crest of Thailand HIV-1 gp120 is proposed to be the most probable conformation responsible for the formation of viral antigen-antibody complex in particular case under study. PMID- 12529160 TI - Acute sinusitis in children: current treatment strategies. AB - Acute sinusitis is a very common infection in childhood, but its management remains a controversial issue. Antibacterials may be effective in selected children, but direct evidence is limited. One randomized, placebo-controlled trial has shown that amoxicillin or amoxicillin/clavulanate are better than placebo for children with symptoms of nasal discharge and cough that are persistent (over 10 days) and not improving. However, another placebo-controlled trial of the same agents did not demonstrate any benefit from antibacterials in a patient population selected with a clinical diagnosis of sinusitis of moderate severity, based on a composite clinical symptom score. A systematic assessment of cure rates with various antibacterials shows no consistent differences between classes. Evidence on the use of ancillary measures and nasal corticosteroids is also limited. The only randomized, placebo-controlled trial of antihistamines and decongestants has shown no incremental benefit when given in addition to amoxicillin. Another placebo-controlled randomized trial showed some transient symptomatic improvement with the use of nasal corticosteroids. No randomized trials exist on the use of antral lavage in children with acute sinusitis. The current rates of antimicrobial resistance among commonly implicated pathogens should be considered in therapeutic decisions. However, there is no evidence from well-designed trials on specifically how to manage children at high risk of carrying resistant organisms. The inaccuracy of clinical signs and symptoms in documenting the diagnosis further complicates therapeutic decisions. Nevertheless, radiographic assessment does not meaningfully improve the accuracy of the diagnosis for uncomplicated cases, and it is not cost effective. In the absence of definitive evidence, treatment with amoxicillin 45 mg/kg/day in two divided doses may be used in selected patients with symptoms that are persistent and not improving. High doses (90 mg/kg in two divided doses) may also be considered, and amoxicillin/clavulanate may be a more appropriate choice when there is high risk of resistant pathogens, e.g. in a child attending a childcare center, or recent use of antibacterials. However, a considerable proportion of children, especially those with mild or improving symptoms, may not have to be treated at all. PMID- 12529162 TI - Graves disease in childhood: a review of the options for diagnosis and treatment. AB - While diagnosing Graves disease in childhood and adolescence does not usually present specific problems, the treatment of hyperthyroidism is still controversial. In particular, with regard to the use of radioiodine therapy, strategies vary between many European and North American pediatric endocrinology centers. After the diagnosis is made, antithyroid drug treatment with methimazole (thiamazole), carbimazole, or propylthiouracil should be performed with caution, in particular, because of severe adverse effects, such as agranulocytosis or hepatitis, that are found in up to 1% of patients. Antithyroid drug treatment should not be continued long-term, particularly since definitive remission cannot be expected in more than 30-40% of patients. In contrast, the risk of severe adverse effects is still present, and the risk of thyroid carcinoma increases with time and appears to be considerably higher than after radioiodine treatment. To a great extent, the success of surgery depends on the skills of a trained surgeon. The question of whether to perform total or subtotal thyroidectomy is yet to be resolved. Surgery should be considered in patients with a large thyroid gland (>80g), severe ophthalmopathy, and a lack of remission on antithyroid drug treatment. Success rates have increased to up to 97%, while severe adverse effects (laryngeal nerve palsy, hypoparathyroidism) occur in approximately 4% of patients. Mortality is below 0.1%. Radioiodine treatment in children >5 years of age does not appear to be associated with an increased risk of thyroid carcinoma; however, long-term data are lacking. Compared with the surgical approach, success rates are lower, particularly if low doses of radioiodine are used. In general, adverse effects are less prevalent than in patients who have undergone surgery. PMID- 12529161 TI - Therapy for acute rejection in pediatric organ transplant recipients. AB - Despite the availability of potent immunosuppressive drugs, rejection after organ transplantation in children remains a serious concern, and may lead to significant morbidity, graft loss, and death of the patient. Acute graft rejection in pediatric recipients is first treated with methylprednisolone pulses, followed by progressive taper of corticosteroid doses. After control of the rejection episode, baseline immunosuppression has to be adjusted and closely monitored since rejection (especially late episodes, occurring more than 6 months after transplantation) may be due to a lack of compliance or sub-therapeutic drug concentrations. The management of corticosteroid resistant rejection is not standardized, and depends on the transplanted organ and previous immunosuppressive regimen. In patients experiencing corticosteroid resistant acute rejection while on a cyclosporine-based immunosuppressive regimen, cyclosporine is generally changed to tacrolimus. In case of tacrolimus-based immunosuppression, tacrolimus blood levels may be increased, and/or mycophenolate mofetil (which nowadays tends to replace azathioprine) or sirolimus may be added, although pharmacodynamic data and clinical studies with these agents are still scarce in pediatric recipients. The use of antithymocyte globulins or monoclonal anti-CD3 antibodies, muromonab CD3 (OKT3) is hampered by numerous adverse effects, including a significant risk of over-immunosuppression. These therapies are nowadays indicated in very selected cases. Other treatments such as plasmapheresis and high dose immunoglobulins may be useful in difficult cases. In patients with refractory rejection despite therapeutic escalation, the risks of over-immunosuppression, including opportunistic infections and malignancies (especially the Epstein-Barr virus related post-transplant lymphoproliferative disease) have to be balanced with the consequences of graft loss due to rejection. Detransplantation or retransplantation may, in some instances, be preferable to severe infectious or tumoral complications. PMID- 12529164 TI - Neuraminidase inhibitors in pediatric patients: potential place in influenza therapy. AB - Influenza is responsible for annual epidemics, and is associated with significant morbidity and mortality. The development and use of antiviral agents is one of the recent strategies used to control influenza. Zanamivir and oseltamivir are members of a new class of antiviral agents, the neuraminidase inhibitors, that are effective in the treatment of influenza. These drugs have been evaluated in double-blind, placebo-controlled trials in children. Efficacy was shown by shortened duration of influenza-related symptoms in children treated with neuraminidase inhibitors. Seasonal prophylaxis in adults, and administration to household contacts of influenza-infected persons has decreased the incidence of influenza infection and illness. Adverse effects of zanamivir are minimal, while oseltamivir is associated with gastrointestinal upset, which does not usually require the medication to be ceased. No serious adverse events have been reported in children receiving these medications. Caution should be exercised in the use of zanamivir in children with asthma due to the potential development of bronchospasm and respiratory distress. Development of resistance to these medications is reported to occur less frequently than with amantadine. Resistance to zanamivir has been rarely detected during clinical trials, while up to 5.5% of influenza strains isolated from children after treatment were reported to be resistant to oseltamivir. Due to the role children play in the spread of influenza within the community, use of these medications may help control influenza epidemics. PMID- 12529165 TI - Shortened course of antibacterial therapy for acute otitis media. AB - Acute otitis media (AOM) is the most frequent bacterial disease and leading cause of antibacterial prescriptions in infants and children. Although AOM resolves spontaneously in most patients, antibacterial treatment is believed to be indicated in most industrial countries, particularly for infants, to prevent severe complications and relieve symptoms more rapidly. The classical duration of oral antibacterial therapy has been 10 days. During the last decade, the efficacy of shortened course antibacterial therapy has been demonstrated in many trials. Advantages of shorter treatment periods include less expense, better compliance, and potentially less impact on the commensal flora. However, short course antibacterial therapy may not be appropriate for children <2 years of age, particularly those attending daycare centers, those with otorrhea or a recent episode (<1 month), and otitis-prone children. Reasons for a poorer efficacy in these groups of children are still not completely understood. PMID- 12529166 TI - Detection and analysis of urinary peptides by on-line liquid chromatography and mass spectrometry: application to patients with renal Fanconi syndrome. AB - Urinary proteomics has become a topical and potentially valuable field of study in relation to normal and abnormal renal function. Filtered bioactive peptides present in high concentration in the nephron of patients with tubular proteinuria may have downstream effects on renal tubular function. In renal Fanconi syndromes, such as Dent's disease, peptides implicated in altered tubular function or injury have recently been measured in urine by immunochemical methods. However, the limited availability of antibodies means that only certain peptides can be detected in this way. We have used nanoflow liquid chromatography and tandem mass spectrometry (nanoLC-MS/MS) as a complementary technique to analyse urinary peptides. Urine was desalted by solid-phase extraction (SPE) and its peptides were then separated from neutral and acidic compounds by strong cation-exchange chromatography (SCX), which was also used to fractionate the peptide mixture. Fractions from the SCX step were separated further by reversed phase LC and analysed on-line by MS/MS. Extraction by SPE showed a good recovery of small peptides. We detected over 100 molecular species in urine samples from three individuals with Dent's disease. In addition to plasma and known urinary proteins, we identified some novel proteins and potentially bioactive peptides in urine from these patients, which were not present in normal urine. These data show that nanoLC-MS/MS complements existing techniques for the identification of polypeptides in urine. This approach is a potentially powerful tool to discover new markers and/or causative factors in renal disease; in addition, its sensitivity may also make it applicable to the direct ultramicroanalysis of renal tubule fluid. PMID- 12529163 TI - Nonsteroidal anti-inflammatory drugs for postoperative pain: a focus on children. AB - Pain is a common symptom after surgery in children, and the need for effective pain management is obvious. For example, after myringotomy, despite the brief nature of the procedure, at least one-half of children have significant pain. After more extended surgery, such as tonsillectomy, almost all children have considerable pain longer than 7 days. Nonsteroidal anti-inflammatory drugs (NSAIDs) are useful for postoperative pain management because surgery causes both pain and inflammation. Several pediatric studies indicate NSAIDs are effective analgesics in the management of mild and moderate pain. In the treatment of severe pain, NSAIDs should be given with acetaminophen (paracetamol) or opioids, and the use of an appropriate regional analgesic technique should be considered. NSAIDs are more effective in preventing pain than in the relief of established pain. Pain following surgery is best managed by providing medication on a regular basis, preventing the pain from recurring. This proactive approach should be implemented for any procedure where postoperative pain is the likely outcome. In children, the choice of formulation can be more important than the choice of drug. Intravenous administration is preferred for children with an intravenous line in place; thereafter mixtures and small tablets are feasible options. Children dislike suppositories, and intramuscular administration should not be used in nonsedated children. Ibuprofen, diclofenac, ketoprofen and ketorolac are the most extensively evaluated NSAIDs in children. Only a few trials have compared different NSAIDs, but no major differences in the analgesic action are expected when appropriate doses of each drug are used. Whether NSAIDs differ in the incidence and severity of adverse effects is open to discussion. Because NSAIDs prevent platelet aggregation they may increase bleeding. A few studies indicate that ketorolac may increase bleeding more so than other NSAIDs, but the evidence is conflicting. Severe adverse effects of NSAIDs in children are very rare, but it is important to know about adverse effects in order to recognize and treat them when they do occur. NSAIDs are contraindicated in patients in whom sensitivity reactions are precipitated by aspirin (acetylsalicylic acid) or other NSAIDs. They should be used with caution in children with liver dysfunction, impaired renal function, hypovolemia or hypotension, coagulation disorders, thrombocytopenia, or active bleeding from any cause. In contrast, it seems that most children with mild asthma may use NSAIDs. PMID- 12529167 TI - Apolipoprotein E gene polymorphism and colorectal cancer: gender-specific modulation of risk and prognosis. AB - Apolipoprotein E ( ApoE ) gene polymorphism is a major factor in lipid metabolism. It has been suggested that this polymorphism can modulate colorectal tumour risk. We tested this hypothesis for colorectal cancer (CRC). ApoE genotype was determined in 206 patients with CRC and 353 healthy controls from the East Anglia region of the U.K. Compared with individuals possessing the most common epsilon 3/epsilon 3 genotype, those with the epsilon 2/epsilon 3 genotype had an increased risk of colon cancer [odds ratio (OR) = 1.91; 95% confidence interval 1.05-3.45]. However, this association was strongly affected by gender. Separate analysis of male and female subjects revealed a highly significant association in men (OR = 2.71; 95% confidence interval 1.30-5.65), but no association in women (OR = 1.01; 95% confidence interval 0.37-2.77). Likewise, the proportion of male patients with more advanced tumours (Dukes' C&D) was significantly increased among those with the ApoE epsilon 2/epsilon 3 genotype (OR = 4.16; 95% confidence interval 1.36-12.75). No significant effect of the presence of the epsilon 4 allele on CRC risk was found; however, there were no epsilon 4/epsilon 4 homozygotes among patients with proximal colon cancers. The ApoE epsilon 3/epsilon 3 majority genotype appeared to be associated with the lowest risk of CRC. These results suggest that ApoE genotype can influence both CRC risk and prognosis of the existing disease in a gender-dependent manner. PMID- 12529168 TI - Influence of leg position and environmental temperature on segmental volume expansion during venous occlusion plethysmography. AB - Blood flow determinations by venous occlusion plethysmography applying the strain gauge technique are frequently used. A problem with the strain-gauge technique is that the relationship between venous volume and transmural pressure is not linear and, furthermore, changes with the sympathetic tone. The present study tests the hypothesis that these factors lead to a redistribution of venous blood, which may impair the accuracy of the technique. The relative volume expansion rates of four leg segments were studied with the leg in different positions and at disparate temperatures, thereby inducing varying venous pressures and sympathetic tone ( n =6). With elevated leg and relaxed veins (at 50 degrees C), the distal thigh showed a relatively low expansion rate (25.8+/-4.5 ml.min(-1).l(-1)), whereas values in the calf segments were higher (34.5-39.0 ml.min(-1).l(-1)). With lower initial transmural pressure, calf segments can increase their volume much more during occlusion compared with the distal thigh. In a higher transmural pressure region (lowered leg), the difference in compliance between limb segments is less. In this case, compliance and volume expansion rate was higher in the distal thigh (14.2, 13.5 and 22.2 ml.min(-1).l(-1) at 10, 20 and 50 degrees C respectively) than in the calf segments (for the distal calf: 6.4, 7.7 and 16.2 ml.min(-1).l( 1) respectively). There was a significant interaction ( P <0.001) between temperature and leg position, indicating a higher degree of sympathetic vasoactivity in the calf. It is concluded that blood flow determination by strain gauge plethysmography is less accurate, due to a potential redistribution of the venous blood. Therefore possible influences of variations in sympathetic tone and venous pressure must be considered even in intra-individual comparisons, especially in interventional studies. PMID- 12529169 TI - Nitrogen-source regulation of yeast gamma-glutamyl transpeptidase synthesis involves the regulatory network including the GATA zinc-finger factors Gln3, Nil1/Gat1 and Gzf3. AB - In Saccharomyces cerevisiae, the CIS2 gene encodes gamma-glutamyl transpeptidase (gamma-GT; EC 2.3.2.2), the main GSH-degrading enzyme. The promoter region of CIS2 contains one stress-response element (CCCCT) and eight GAT(T/A)A core sequences, probably involved in nitrogen-regulated transcription. We show in the present study that expression of CIS2 is indeed regulated according to the nature of the nitrogen source. Expression is highest in cells growing on a poor nitrogen source such as urea. Under these conditions, the GATA zinc-finger transcription factors Nil1 and Gln3 are both required for CIS2 expression, Nil1 appearing as the more important factor. We further show that Gzf3, another GATA zinc-finger protein, acts as a negative regulator in nitrogen-source control of CIS2 expression. During growth on a preferred nitrogen source like NH(4)(+), CIS2 expression is repressed through a mechanism involving (at least) the Gln3-binding protein Ure2/GdhCR. Induction of CIS2 expression during nitrogen starvation is dependent on Gln3 and Nil1. Furthermore, rapamycin causes similar CIS2 activation, indicating that the target of rapamycin signalling pathway controls CIS2 expression via Gln3 and Nil1 in nitrogen-starved cells. Finally, our results show that CIS2 expression is induced mainly by nitrogen starvation but apparently not by other types of stress. PMID- 12529170 TI - A pheromone-binding protein from the cockroach Leucophaea maderae: cloning, expression and pheromone binding. AB - Odorant-binding proteins (OBPs) are thought to transport volatile compounds from air to their receptors through the sensillary lymph. In this protein family, the subgroup of pheromone-binding proteins (PBPs) is specifically tuned to the perception of the sexual pheromone. To date, the description of OBPs has been restricted to Endopterygota and Paraneoptera. Their expression in Orthopteroid has been hypothesized, but no evidence of OBP has been produced in this assemblage to date. In the present study, we describe the first OBP from a Dictyopteran insect that belongs to the cockroach Leucophaea maderae. The PBP of L. maderae (PBPLma) shares all the hallmarks of the OBP family and is expressed specifically in the female adult antennae, the sex that perceives the sexual pheromone. The affinity of the recombinant PBPLma produced in the Escherichia coli periplasm for the pheromonal compounds has been tested by displacement of a fluorophore, 8-anilino-1-naphtalenesulphonic acid (ANS). Our results suggest that two chemically close compounds of the pheromonal blend (3-hydroxy-butan-2-one and butane-2,3-diol) are capable of displacing ANS, whereas two other pheromone components (E-2-octenoic acid and senecioic acid) and other alkyl volatile compounds are not capable of displacing ANS, indicating a certain filtering of binding, which can be correlated with the putative function. PMID- 12529171 TI - Alkaline transition of phytocyanins: a comparison of stellacyanin and umecyanin. AB - The effect of pH on Cu(I) and Cu(II) forms of the isolated soluble domain of the stellacyanin from Rhus vernicifera (SCu), the Japanese lacquer tree, has been studied by electronic and NMR spectroscopy and using direct electrochemical measurements. A pK(a) value of 10.1-10.4 is observed for the alkaline transition in this oxidized phytocyanin and results in a slightly altered active-site structure, as indicated by changes in the visible and paramagnetic (1)H NMR spectra. Electrochemical studies show that the pK(a) value for this transition in SCu(I) (reduced SCu) is 11.0. These results are compared with those recently obtained for other phytocyanins, and in particular umecyanin. In all cases, the alkaline transition is caused by the deprotonation of the surface lysine residue adjacent to the axial ligand. This lysine residue is completely conserved in known phytocyanin sequences. Also highlighted in these studies are the remarkable active-site similarities between stellacyanin and umecyanin. PMID- 12529172 TI - Docking sites on mitogen-activated protein kinase (MAPK) kinases, MAPK phosphatases and the Elk-1 transcription factor compete for MAPK binding and are crucial for enzymic activity. AB - Mitogen-activated protein kinase (MAPK) cascades control gene expression patterns in response to extracellular stimuli. MAPK/ERK (extracellular-signal-regulated kinase) kinases (MEKs) activate MAPKs by phosphorylating them; activated MAPKs, in turn, phosphorylate target transcription factors, and are deactivated by phosphatases. One mechanism for maintaining signal specificity and efficiency is the interaction of MAPKs with their substrates and regulators through high affinity docking sites. In the present study, we show that peptides corresponding to the MAPK-docking sites of MEK1, MEK2, Ste7, Elk-1 and MAPK phosphatase (MKP)-2 potently inhibit MEK2 phosphorylation of ERK2, ERK2 phosphorylation of Elk-1, and MKP-1 dephosphorylation of ERK2. Each peptide inhibited multiple reactions; for example, the MEK2 peptide inhibited not only MEK2, but also ERK2 and MKP-1. In addition, these docking-site peptides inhibited MEK2-ERK2 binding. The MAPK docking site of MEK1 also potently stimulated ERK2-mediated phosphorylation of a target site on the same peptide. Control peptides with mutations of conserved basic and hydrophobic residues of the MAPK-docking site consensus lacked biological activity. We conclude that MEKs, MKPs and the Elk-1 transcription factor compete for binding to the same region of ERK2 via protein-protein interactions that are crucial for kinase/phosphatase activity. PMID- 12529173 TI - The Fn14 cytoplasmic tail binds tumour-necrosis-factor-receptor-associated factors 1, 2, 3 and 5 and mediates nuclear factor-kappaB activation. AB - Fn14 is a growth-factor-inducible immediate-early-response gene encoding a 102 amino-acid type I transmembrane protein. The human Fn14 protein was recently identified as a cell-surface receptor for the tumour necrosis factor (TNF) superfamily member named TWEAK (TNF-like weak inducer of apoptosis). In the present paper, we report that the human TWEAK extracellular domain can also bind the murine Fn14 protein. Furthermore, site-specific mutagenesis and directed yeast two-hybrid interaction assays revealed that the TNFR-associated factor (TRAF) 1, 2, 3 and 5 adaptor molecules bind the murine Fn14 cytoplasmic tail at an overlapping, but non-identical, amino acid sequence motif. We also found that TWEAK treatment of quiescent NIH 3T3 cells stimulates inhibitory kappaBalpha phosphorylation and transcriptional activation of a nuclear factor-kappaB (NF kappaB) enhancer/luciferase reporter construct. Fn14 overexpression in transiently transfected NIH 3T3 cells also promotes NF-kappaB activation, and this cellular response requires an intact TRAF binding site. These results indicate that Fn14 is a functional TWEAK receptor that can associate with four distinct TRAF family members and stimulate the NF-kappaB transcription factor signalling pathway. PMID- 12529174 TI - Protein-protein interactions involved in the recognition of p27 by E3 ubiquitin ligase. AB - The p27(Kip1) protein is a potent cyclin-dependent kinase inhibitor, the level of which is decreased in many common human cancers as a result of enhanced ubiquitin dependent degradation. The multiprotein complex SCF(Skp2) has been identified as the ubiquitin ligase that targets p27, but the functional interactions within this complex are not well understood. One component, the F-box protein Skp2, binds p27 when the latter is phosphorylated on Thr(187), thus providing substrate specificity for the ligase. Recently, we and others have shown that the small cell cycle regulatory protein Cks1 plays a critical role in p27 ubiquitination by increasing the binding affinity of Skp2 for p27. Here we report the development of a homogeneous time-resolved fluorescence assay that allows the quantification of the molecular interactions between human recombinant Skp2, Cks1 and a p27 derived peptide phosphorylated on Thr(187). Using this assay, we have determined the dissociation constant of the Skp2-Cks1 complex (K(d) 140 +/- 14 nM) and have shown that Skp2 binds phosphorylated p27 peptide with high affinity only in the presence of Cks1 (K(d) 37 +/- 2 nM). Cks1 does not bind directly to the p27 phosphopeptide or to Skp1, which confirms its suggested role as an allosteric effector of Skp2. PMID- 12529175 TI - Catalytic properties and inhibition of proline-specific dipeptidyl peptidases II, IV and VII. AB - There is currently intense interest in the emerging group of proline-specific dipeptidases, and their roles in the regulation of biological processes. Dipeptidyl peptidase IV (DPP-IV) is involved in glucose metabolism by contributing to the regulation of glucagon family peptides and has emerged as a potential target for the treatment of metabolic diseases. Two other proline specific dipeptidases, DPP-VII (also known as quiescent cell proline dipeptidase) and DPP-II, have unknown functions and have recently been suggested to be identical proteases based on a sequence comparison of human DPP-VII and rat DPP II (78% identity) [Araki, Li, Yamamoto, Haneda, Nishi, Kikkawa and Ohkubo (2001) J. Biochem. 129, 279-288; Fukasawa, Fukasawa, Higaki, Shiina, Ohno, Ito, Otogoto and Ota (2001) Biochem. J. 353, 283-290]. To facilitate the identification of selective substrates and inhibitors for these enzymes, a complete biochemical profile of these enzymes was obtained. The pH profiles, substrate specificities as determined by positional scanning, Michaelis-Menten constants and inhibition profiles for DPP-VII and DPP-II were shown to be virtually identical, strongly supporting the hypothesis that they are the same protease. In addition, substrate specificities, catalytic constants and IC(50) values were shown to be markedly different from those of DPP-IV. Selective DPP-IV and DPP-VII substrates were identified and they can be used to design selective inhibitors and probe further into the biology of these enzymes. PMID- 12529176 TI - Neurogenin3 triggers beta-cell differentiation of retinoic acid-derived endoderm cells. AB - Neurogenin3 is a member of the basic helix-loop-helix ('bHLH') family of transcription factors. It plays a crucial role in the commitment of embryonic endoderm into the pancreatic differentiation programme. This factor is considered to act upstream of a cascade of other transcription factors, leading to the fully differentiated endocrine phenotype. Direct observation of the sequential activation of these factors starting from Neurogenin3 had never been demonstrated. By using retinoic acid-derived-endoderm F9 cells as a model, the present study indicates that the ectopic expression of Neurogenin3 is able to start the differentiation pathway of endocrine pancreas. Neurogenin3 triggers the expression of several pancreatic transcription factors following a well defined temporal activation sequence. By reverse transcriptase PCR, immunohistochemistry and RIA, it is shown that stable transfected cells are able to form embryod bodies that produce insulin in response to glucose stimulation. This is the first report of a differentiation event induced by the ectopic expression of a transcription factor in embryonic pluripotent stem cells. PMID- 12529177 TI - Osmotic regulation of insulin-induced mitogen-activated protein kinase phosphatase (MKP-1) expression in H4IIE rat hepatoma cells. AB - A contribution of intracellular dehydration to insulin resistance has been established in human subjects and in different experimental systems. Here the effect of hyperosmolarity (405 mosmol/l) on insulin-induced mitogen-activated protein (MAP) kinase phosphatase (MKP)-1 expression was studied in H4IIE rat hepatoma cells. Insulin induces robust MKP-1 expression which correlates with a vanadate-sensitive decay of extracellular-signal-regulated kinase (Erk-1/Erk-2) activity. Hyperosmolarity delays MKP-1 accumulation by insulin and this corresponds to impaired MKP-1 synthesis, whereas MKP-1 degradation remains unaffected by hyperosmolarity. Rapamycin, which inhibits signalling downstream from the mammalian target of rapamycin (mTOR) and a peptide inhibiting protein kinase C (PKC) zeta/lambda abolish insulin-induced MKP-1 protein but not mRNA expression, suggesting the involvement of the p70 ribosomal S6 protein kinase (p70S6-kinase) and/or the eukaryotic initiation factor 4E-binding proteins (4E BPs) as well as atypical PKCs in MKP-1 translation. Hyperosmolarity induces sustained suppression of p70S6-kinase and 4E-BP1 hyperphosphorylation by insulin, whereas insulin-induced tyrosine phosphorylation of the insulin receptor (IR) beta subunit and the IR substrates IRS1 and IRS2, recruitment of the phosphoinositide 3-kinase (PI 3-kinase) regulatory subunit p85 to the receptor substrates as well as PI 3-kinase activation, and Ser-473 phosphorylation of protein kinase B and Thr-410/403 phosphorylation of PKC zeta/lambda are largely unaffected under hyperosmotic conditions. The hyperosmotic impairment of both, MKP-1 expression and p70S6-kinase hyperphosphorylation by insulin is insensitive to K(2)CrO(4), calyculin A and vanadate, and inhibition of the Erk-1/Erk-2 and p38 pathways. The suppression of MKP-1 may further contribute to insulin resistance under dehydrating conditions by allowing unbalanced MAP kinase activation. PMID- 12529178 TI - Inhibition of oxygen evolution in Photosystem II by Cu(II) ions is associated with oxidation of cytochrome b559. AB - We have found that elevated copper concentrations, apart from the inhibition of oxygen evolution, changed the initial states distribution of the oxygen-evolving complex. Already at low concentrations, copper ions oxidized the low-potential form of cytochrome b (559) and also its high-potential form at higher concentrations at which fluorescence quenching was observed. We suggest that the primary target sites in Photosystem II for copper is tyrosine(z), both cytochrome b (559) forms and chlorophyll(z), and that these sites are the source of the copper-induced fluorescence quenching and oxygen evolution inhibition in Photosystem II. PMID- 12529179 TI - Electrostatic compared with hydrophobic interactions between bovine serum amine oxidase and its substrates. AB - A steady-state kinetic study of bovine serum amine oxidase activity was performed, over a wide range of pH values (5.4-10.2) and ionic strength (10-200 mM), using various (physiological and analogue) substrates as specific probes of the active-site binding region. Relatively small changes in k (cat) values (approx. one order of magnitude) accompanied by marked changes in K(m) and k(cat)/K(m) values (approx. six orders of magnitude) were observed. This behaviour was correlated with the presence of positively charged groups or apolar chains in the substrates. In particular, it was found that the docking of the physiological polyamines, i.e. spermidine and spermine, appears to be modulated by three amino acid residues of the active site, which we have named L(-)H(+), G( )H(+) and IH(+), characterized by p K (a) values of 6.2+/-0.2 [Di Paolo, Scarpa, Corazza, Stevanato and Rigo (2002) Biophys. J. 83, 2231-2239], 8.2+/-0.3 and 7.8+/-0.4 respectively. The electrostatic interaction between the protonated substrates and the enzyme containing the residues L(-)H(+), G(-)H(+) and IH(+) in the deprotonated form, the on/off role of the IH(+) residue and the role of hydrophobic interactions with substrates characterized by apolar chains are discussed. PMID- 12529180 TI - Purification and properties of a transglutaminase produced by a Bacillus circulans strain isolated from the Amazon environment. AB - A new microbial transglutaminase (EC 2.3.2.13) from a Bacillus circulans strain isolated from the aquatic Amazonian environment was purified and characterized. Enzyme purification started with (NH(4))(2)SO(4) 'salting out' and proceeded with liquid chromatography on Q-Sepharose FF and octyl-Sepharose 4 FF. The purification factor was approx. 150-fold with a yield of 32%. The enzyme's molecular mass was estimated as 45000 Da on SDS/PAGE. The purified transglutaminase had an optimum temperature of 47 degrees C, the optimum pH of the reaction was 7 and it presented no calcium-dependent activity. PMID- 12529181 TI - Would you be surprised if this patient died?: Preliminary exploration of first and second year residents' approach to care decisions in critically ill patients. AB - BACKGROUND: How physicians approach decision-making when caring for critically ill patients is poorly understood. This study aims to explore how residents think about prognosis and approach care decisions when caring for seriously ill, hospitalized patients. METHODS: Qualitative study where we conducted structured discussions with first and second year internal medicine residents (n = 8) caring for critically ill patients during Medical Intensive Care Unit Ethics and Discharge Planning Rounds. Residents were asked to respond to questions beginning with "Would you be surprised if this patient died?" RESULTS: An equal number of residents responded that they would (n = 4) or would not (n = 4) be surprised if their patient died. Reasons for being surprised included the rapid onset of an acute illness, reversible disease, improving clinical course and the patient's prior survival under similar circumstances. Residents reported no surprise with worsening clinical course. Based on the realization that their patient might die, residents cited potential changes in management that included clarifying treatment goals, improving communication with families, spending more time with patients and ordering fewer laboratory tests. Perceived or implied barriers to changes in management included limited time, competing clinical priorities, "not knowing" a patient, limited knowledge and experience, presence of diagnostic or prognostic uncertainty and unclear treatment goals. CONCLUSIONS: These junior level residents appear to rely on clinical course, among other factors, when assessing prognosis and the possibility for death in severely ill patients. Further investigation is needed to understand how these factors impact decision making and whether perceived barriers to changes in patient management influence approaches to care. PMID- 12529182 TI - Attitudes of developing world physicians to where medical research is performed and reported. AB - BACKGROUND: Little is known about the influence of the site of research or publication on the impact of the research findings on clinical practice, particularly in developing countries. The International Clinical Epidemiology Network (INCLEN) is dedicated to improving the quality of health research in the Developing World through institutional capacity building for evidence based medicine, and provided the opportunity to examine the likely impact of research location and journal location on physicians' practice in a number of the participating countries. METHODS: Physicians from secondary and tertiary hospitals in six cities located in China, Thailand, India, Egypt and Kenya were enrolled in a cross-sectional questionnaire survey. The primary outcome measures were scores on a Likert scale reflecting stated likelihood of changing clinical practice depending on the source of the research or its publication. RESULTS: Overall, local research and publications were most likely to effect change in clinical practice, followed by North American, European and regional research/publications respectively, although there were significant variations between countries. The impact of local and regional research would be greater if the perceived research quality improved in those settings. CONCLUSION: Conducting high quality local research is likely to be an effective way of getting research findings into practice in developing countries. PMID- 12529183 TI - Frequency of CHEK2*1100delC in New York breast cancer cases and controls. AB - BACKGROUND: The 1100delC CHEK2 allele has been associated with a 1.4-4.7 fold increased risk for breast cancer in women carrying this mutation. While the frequency of 1100delC was 1.1-1.4% in healthy Finnish controls, the frequency of this allele in a North American control population and in North American breast cancer kindreds remains unclear. METHODS: We genotyped 1665 healthy New York volunteers and 300 cases of breast cancer for the CHEK2*1100delC. RESULTS: The overall frequency of the 1100delC was 3/300 (1.0%) among all cases with either a family history of breast cancer (n = 192) or a personal history of breast cancer (n = 108, of which 46 were bilateral, 46 unilateral, and 16 were male breast cancer cases), compared to a frequency of 5/1665 (0.3%) in healthy controls (p = 0.1). There was no difference in allele frequency among Ashkenazi and non Ashkenazi controls. CONCLUSION: The relatively low breast cancer penetrance of this allele, along with the low population frequency, will limit the clinical applicability of germline testing for CHEK2*1100delC in North American kindreds. PMID- 12529184 TI - Comparative study of methyl-CpG-binding domain proteins. AB - BACKGROUND: Methylation at CpG dinucleotides in genomic DNA is a fundamental epigenetic mechanism of gene expression control in vertebrates. Proteins with a methyl-CpG-binding domain (MBD) can bind to single methylated CpGs and most of them are involved in transcription control. So far, five vertebrate MBD proteins have been described as MBD family members: MBD1, MBD2, MBD3, MBD4 and MECP2. RESULTS: We performed database searches for new proteins containing an MBD and identified six amino acid sequences which are different from the previously described ones. Here we present a comparison of their MBD sequences, additional protein motifs and the expression of the encoding genes. A calculated unrooted dendrogram indicates the existence of at least four different groups of MBDs within these proteins. Two of these polypeptides, KIAA1461 and KIAA1887, were only present as predicted amino acid sequences based on a partial human cDNA. We investigated their expression by Northern blot analysis and found transcripts of ~8 kb and ~5 kb respectively, in all eight normal tissues studied. CONCLUSIONS: Eleven polypeptides with a MBD could be identified in mouse and man. The analysis of protein domains suggests a role in transcriptional regulation for most of them. The knowledge of additional existing MBD proteins and their expression pattern is important in the context of Rett syndrome. PMID- 12529186 TI - The worldwide epidemic of obesity in adolescents. AB - There is an epidemic of obesity affecting adolescents worldwide. Both in developing and in developed countries, adolescents are increasingly becoming more obese. The number of adolescents exceeding previously identified cut-points as well as the weight and fatness of the most obese individuals is increasing at a progressive rate. Despite their benign appearance, epidemics of noncommunicable disease (or their risk factors) are no less devastating to the health of populations. The two key responses to any epidemic are to discover the causes of the epidemic disease and to characterize the epidemic. The latter needs to occur in relation to prevalence, distribution across the populations (are some population groups more likely to be affected than others?), and secular trends. This chapter reviews what is currently known about the epidemiology of overweight and obesity among adolescents throughout the world. To clarify terms of epidemiology, this chapter first identifies what are considered the most appropriate measures of adiposity and defines how much fat is too much fat. PMID- 12529185 TI - Optimal step length EM algorithm (OSLEM) for the estimation of haplotype frequency and its application in lipoprotein lipase genotyping. AB - BACKGROUND: Haplotype based linkage disequilibrium (LD) mapping has become a powerful and cost-effective method for performing genetic association studies, particularly in the search for genetic markers in linkage disequilibrium with complex disease loci. Various methods (e.g. Monte-Carlo (Gibbs sampling); EM (expectation maximization); and Clark's method) have been used to estimate haplotype frequencies from routine genotyping data. RESULTS: These algorithms can be very slow for large number of SNPs. In order to speed them up, we have developed a new algorithm using numerical analysis technology, a so-called optimal step length EM (OSLEM) that accelerates the calculation. By optimizing approximately the step length of the EM algorithm, OSLEM can run at about twice the speed of EM. This algorithm has been used for lipoprotein lipase (LPL) genotyping analysis. CONCLUSIONS: This new optimal step length EM (OSLEM) algorithm can accelerate the calculation for haplotype frequency estimation for genotyping data without pedigree information. An OSLEM on-line server is available, as well as a free downloadable version. PMID- 12529187 TI - Medical nutrition therapy of overweight adolescents. AB - The long-term goal of medical nutrition therapy for adolescents who are overweight or at risk for becoming overweight is to promote healthy lifestyle behaviors. These behaviors will, in turn, improve metabolic parameters and self esteem while helping the adolescent achieve and maintain a desirable body weight. The identification of anthropometric, metabolic, nutritional, and environmental risk factors present in the child and family will help formulate the medical nutrition intervention. A well-balanced diet that supports growth and development, aerobic exercise, and cognitive behavioral strategies are essential components of an intervention program. Frequent and long-term monitoring by a registered dietitian and pediatrician will reinforce lifestyle changes and support the adolescent and family in achieving realistic goals of weight loss or weight maintenance. This article covers the assessment and interventions necessary for successful nutrition therapy for obese and superobese adolescents. PMID- 12529188 TI - Physical activity, exercise, and sedentary activity: relationship to the causes and treatment of obesity. AB - Lack of physical activity is a known determinant of obesity. Unfortunately, children become less and less active as they reach and progress through adolescence. This behavior leads to detrimental effects on body composition and other components of health-related physical fitness. Various types of exercise and activities have different effects on fitness, body composition, and metabolic state. For a physician to recommend activities or sports and exercise type, he or she should examine and discuss the patients fitness and physique goals to maintain their enthusiasm and promote lifelong physical activity benefits. PMID- 12529189 TI - Cognitive-behavioral approaches in the management of obesity. AB - Many of the behavioral interventions designed to promote dietary change in individuals include medical assessment, initial assessment of diet history, assessing readiness, establishing dietary goals, self-monitoring, stimulus control training, training in problem solving, relapse prevention training, enlisting social support, nutrition education, dietary therapy, and ongoing contact to maintain progress. The comprehensive nature of a cognitive-behavioral weight management program is of value in modifying behaviors that are linked to adverse health effects and psychological distresses, without necessarily causing a drastic weight loss in obese individuals. The behavioral treatments for overweight and obesity directly modify behaviors that bear on health and illness, such as improving dietary choices, decreasing sedentary behaviors, and increasing habitual physical activity and exercises. Cognitive-behavioral treatment can be used to help overweight adolescents become more assertive in coping with the adverse social stigma of being overweight, enhance their self-esteem, and reduce their dissatisfaction with body image regardless of their weight loss. Cognitive behavioral treatments seem to be more effective in children when delivered before puberty than they are for adults. PMID- 12529190 TI - Eating disorders in children and young adolescents: etiology, classification, clinical features, and treatment. AB - Eating disorders in children and young adolescents are increasing in prevalence and occurring at ever-earlier ages. Eating disorders in this age group differ from the traditional eating disorders seen in older adolescents and young adults in that they frequently present atypically, they are suspected less often, they are more difficult to diagnose definitively, and there is less evidence to guide informed treatment decisions. Classification of eating disorders in this population also presents unique challenges. A high index of suspicion and aggressive screening will help to prevent delays in diagnosis. Treatment should include attention to nutritional, medical, and psychological issues. Family therapy has emerged as the treatment modality with the greatest evidence for its efficacy. Prognosis has been poorly studied, but good outcomes are common, particularly with early intervention. PMID- 12529191 TI - Psychosexual development and eating disorders. AB - The relationship among eating disorders (EDs), psychosexual and identity development, and physical maturation (puberty) is reviewed. The developmental tasks of adolescence are summarized, and research from both community studies and clinical samples on the association between the development of an ED and putative risk factors that include pubertal development and psychosexual behaviors and attitudes for children and adolescents is reviewed. Specific issues explored include the role of child and adolescent abuse and EDs in males. Overall evidence suggests the following: there are inconsistent findings regarding early pubertal development as a risk factor; there is some support for differences between the ED subtypes in sexual attitudes, behaviors, and experiences; sexual abuse is not a specific risk factor; and gender identity issues may play more of a role for adolescent males than females. However, psychosexual issues are neither sufficient nor necessary for the development of an ED in a young person. It should be considered as only one factor in the multidimensional, multifactorial framework needed to clarify this complex and still poorly understood set of disorders. PMID- 12529192 TI - The use of indirect calorimetry in the clinical management of adolescents with nutritional disorders. AB - Indirect calorimetry is a noninvasive, inexpensive technique used to determine resting energy expenditure. Its use provides the clinician with objective information that can be used to design, implement, and evaluate efficacy of treatment in the nutritional management of adolescents with anorexia nervosa, bulimia nervosa, chronic dieting behavior, and obesity. This chapter outlines the theoretical framework, interpretation of data, and clinical applications of indirect calorimetry and presents case examples to underscore its utility in adolescents with eating disorders. PMID- 12529193 TI - The female athlete triad. AB - The female athlete triad is a syndrome consisting of disordered eating, amenorrhea, and osteoporosis. The syndrome is increasing in prevalence as more women are participating in sports at a competitive level. Behaviors such as intense exercise or disordered eating patterns can lead to dysregulation of the hypothalamic-pituitarian-ovarian (HPO) axis, resulting in amenorrhea. Hypothalamic amenorrhea can lead to osteoporosis and increased fracture risk. Adolescents may particularly be at risk because it is during this crucial time that females attain their peak bone mass. Prevention of the female athlete triad through education and identification of athletes at risk may decrease the incidence of long-term deleterious consequences. Treatment of the female athlete triad is initially aimed at increasing caloric intake and decreasing physical activity until there is resumption of normal menses. Treatment of decreased bone mineral density and osteoporosis in the adolescent population, however, is controversial, with new treatment modalities currently being investigated in order to aid in the management of this disorder. PMID- 12529194 TI - Osteopenia and osteoporosis in anorexia nervosa. AB - Osteopenia is a frequent and severe complication of anorexia nervosa. Once established, it is difficult to treat and is only partially reversible. Osteoporosis is a preventable disease, and intervention should begin during childhood and adolescence. Optimizing peak bone mass accrual during adolescence is essential, and an episode of anorexia nervosa during adolescence interferes with that process. In anorexia nervosa, results with hormone replacement therapy have been disappointing. Calcium and vitamin D supplementation should be prescribed where necessary. Excessive exercise should be avoided and moderate weight-bearing exercise encouraged. Ongoing research studying newer modalities such as IGF-1, DHEA, and bisphosphonates looks promising. Until more effective treatment regimens become available, the mainstay of treatment remains weight gain, nutritional rehabilitation, and spontaneous resumption of menses. PMID- 12529195 TI - Brain structure and function in adolescents with anorexia nervosa. AB - Anorexia nervosa (AN) commonly arises during adolescence and is associated with significant medical morbidity. Abnormalities in brain structure and function are among the most common, early, and concerning physical consequences. Advances in neuroimaging technology have played an important role in delineating the structural and functional changes found in patients with AN. Studies using computed tomography and magnetic resonance imaging have demonstrated changes in brain structure in the low-weight stages of AN. In addition, functional neuroimaging techniques have demonstrated altered brain metabolism. Debate continues as to whether these brain abnormalities are fully reversible with weight restoration. Neuropsychological research has demonstrated that cognitive dysfunction is also a common feature of AN. Multiple studies have indicated deficits in various neuropsychological domains. Whether the reported cognitive deficits are reversible with weight gain remains unknown. To date, some preliminary evidence suggests that reported cognitive deficits in patients with AN may be associated with structural brain abnormalities. This chapter reviews the current literature about neuroimaging studies and cognitive function in adolescents with AN, discusses the possible underlying mechanisms causing these changes, and explores the possible association between them. PMID- 12529196 TI - Bulimia nervosa and binge-eating disorder in adolescents. AB - Bulimia nervosa (BN) and binge-eating disorder (BED) are separate entities with the common denominator of binge eating. In this chapter, Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV) criteria for BN are reviewed, including both recurrent episodes of binge eating and inappropriate compensatory behaviors to prevent weight gain in one whose self-evaluation is unduly influenced by body weight and shape. Two percent of adolescent females and 0.3% of adolescent males fulfill criteria for BN. Risk factors, medical complications of binge eating (vomiting, use of ipecac, diet pills, diuretics, and laxatives), physical and laboratory findings, and treatment options and outcome are discussed. BED is seen in 1-2% of adolescents. The DSM-IV lists BED under Eating Disorder Not Otherwise Specified. DSM-IV research criteria for BED is reviewed, including binge eating, distress over binge eating, and absence of regular extreme compensatory behaviors. The mean age of onset is 17.2 years. Up to 30% of obese patients have BED. Risk factors are discussed. Because most patients with BED are obese, medical evaluation is similar to that for obesity. Treatment goals must be geared not only toward decreased binge eating but toward weight loss. Outcome is discussed. PMID- 12529197 TI - The role of the primary care practitioner in the treatment of eating disorders. AB - This article addresses practical issues facing the primary care practitioner caring for an adolescent with an eating disorder. It is grounded in the four elements of successful treatment noted by Comerci: (1) recognizing the disorder and restoring physiologic stability early in its course, (2) establishing a trusting, therapeutic partnership with the adolescent, (3) involving the family in treatment, and (4) using an interdisciplinary team approach. Although primary care practitioners often have an established relationship with their patients, adolescents with eating disorders present special challenges. These adolescents tend to be bright, strong-willed, and wary of any recommendations to change their weight-control practices for fear that they will lose control. Their families are often distraught by the conflicts that arise as a result of the disordered eating behaviors and the fear that the condition is associated with significant morbidity and mortality. The article provides primary care clinicians with pragmatic ways to diagnose and initiate treatment and engage the patient and parents as active participants and members of the therapeutic team in the early phases of treatment. In addressing these principles, the authors combine the nurturant-authoritative approach described by Levenkron with the biopsychosocial model proposed by Engel. PMID- 12529198 TI - The course and outcome of eating disorders in adults and in adolescents: a review. AB - Over 100 studies have been published in the literature to date on the course and outcome of eating disorders. These have generally shown that approximately 50% of patients do well over time, approximately 30% do reasonably well but continue to have symptoms, and approximately 20% do poorly. In this article, the literature on the course and outcome of eating disorders is reviewed from two perspectives. The first is an analysis of studies in adults, looking in depth at a range of issues (weight, eating behaviors, menstrual function, psychosocial functioning, psychosexual function, mortality, bulimia nervosa, long-term follow-up, comorbidity, and prognostic factors) and reviewing the difficulties inherent to the performance of all eating disorders follow-up studies. The second is an analysis of whether adolescents with eating disorders have a better prognosis than adults, as has been considered in the literature. It is concluded that adolescents do, in fact, have a somewhat better prognosis than adults, with the differences seeming to be greater with a longer duration of follow-up. PMID- 12529199 TI - Obesity and eating disorder prevention: an integrated approach? AB - This article provides a rationale for interventions aimed at the prevention of eating disorders and obesity, an overview of some of the questions and controversies currently facing the fields of eating disorder and obesity prevention, and a discussion of the potential for integrated prevention approaches that address the broad spectrum of weight-related disorders. A rationale for utilizing an integrated approach, the challenges inherent to developing such an approach, and suggestions for working toward integrated approaches aimed at preventing the broad spectrum of weight-related disorders are discussed. PMID- 12529202 TI - Visual Information and Object Size in the Control of Reaching. AB - The role of vision in the control of reaching and grasping was investigated by varying the available visual information. Adults (N = 7) reached in conditions that had full visual information, visual information about the target object but not the hand or surrounding environment, and no visual information. Four different object diameters were used. The results indicated that as visual information and object size decreased, subjects used longer movement times, had slower speeds, and more asymmetrical hand-speed profiles. Subjects matched grasp aperture to object diameter, but overcompensated with larger grasp apertures when visual information was reduced. Subjects also qualitatively differed in reach kinematics when challenged with reduced visual information or smaller object size. These results emphasize the importance of vision of the target in reaching and show that subjects do not simply scale a command template with task difficulty. PMID- 12529203 TI - The Timing Effects of Accent Production in Periodic Finger-Tapping Sequences. AB - When subjects are required to produce short sequences of equally paced finger taps and to accentuate one of the taps, the interval preceding the forceful tap is shortened and the one that immediately follows the accent is lengthened. Assuming that the tapping movements are triggered by an internal clock, one explanation attributes the rnistiming of the taps to central factors: The momentary rate of the clock is accelerated or decelerated as a function of motor preparation to, respectively, increase or decrease the movement force. This hypothesis predicts that the interresponse intervals measured between either tap movement onsets or movement terminations (taps) will show the same timing pattern. A second explanation for the observed interval effects is that the tapping movements are triggered by a regular internal clock but the timing of the successive taps is altered because the forceful movement is completed in less time than the other tap movements are. This "peripheral" hypothesis predicts regular timing of movement onsets but distorted timing of movement terminations. In the present study, the trajectories of the movements performed by subjects were recorded and the interresponse intervals were measured at the beginning and the end of the tapping movements. The results of Experiment 1 showed that neither model can fully explain the interval effects: The fast forceful movements were initiated with an additional delay that took into account the small execution time of these movements. Experiment 2 reproduced this finding and showed that the timing of the onset and contact intervals did not evolve with the repetition of trial blocks. Therefore, the assumption of an internal clock that would trigger the successive movements must be rejected. The results are discussed in the framework of a modified two-stage model in which the internal clock, instead of triggering the tapping movements, provides target time points at which the movements have to produce their meaningful effects, that is, contacts with the response key. The timing distortions are likely to reflect both peripheral and central components. PMID- 12529204 TI - Should Common Optimal Movement Patterns Be Identified as the Criterion to Be Achieved? AB - Recently, a new paradigm has been proposed for the study of knowledge of performance (Schmidt & Young, 1991). In this paradigm, the experimenters identified an optimal kinematic movement pattern, based on the performance of a best subject, which was imposed on all subjects as the criterion to achieve. This approach, which assumes that this pattern is the best way for all subjects to do the task, was tested in the present experiment. In Experiment 1, a common optimal movement pattern could not be identified by an analysis of scatterplot graphs of the scores plotted as a function of various kinematic vanables or by correlating (within and across subjects [N = 14]) each kinematic variable with the score. In Experiment 2, subjects (N = 12) were retrained in 2 separate sessions, during which they tried to reproduce either a personal best or the best subject's template. Results indicated that scores were similar regardless of the template pattern being used. During training, however, subjects who used their personal template had a lower incidence of zero scores and were more consistent than those who used the template of the best subject. This provided a second line of evidence against the assumption that a common optimal movement pattern exists for this type of task. PMID- 12529205 TI - Response Timing Accuracy as a Function of Movement Velocity and Distance. AB - In two experiments, patterns of response error during a timing accuracy task were investigated. In Experiment 1, these patterns were examined across a full range of movement velocities, which provided a test of the hypothesis that as movement velocity increases, constant error (CE) shifts from a negative to a positive response bias, with the zero CE point occurring at approximately 50% of maximum movement velocity (Hancock & Newell, 1985). Additionally, by examining variable error (VE), timing error variability patterns over a full range of movement velocities were established. Subjects (N = 6) performed a series of forearm flexion movements requiring 19 different movement velocities. Results corroborated previous observations that variability of timing error primarily decreased as movement velocity increased from 6 to 42% of maximum velocity. Additionally, CE data across the velocity spectrum did not support the proposed timing error function. In Experiment 2, the effect(s) of responding at 3 movement distances with 6 movement velocities on response timing error were investigated. VE was significantly lower for the 3 high-velocity movements than for the 3 low velocity movements. Additionally, when MT was mathematically factored out, VE was less at the long movement distance than at the short distance. As in Experiment 1, CE was unaffected by distance or velocity effects and the predicted CE timing error function was not evident. PMID- 12529206 TI - The Scheduling of Knowledge of Results. AB - The experiments outlined in this article were performed so that the acquisition effects of KR scheduling on no-KR retention could be determined. In Experiment 1, the group that alternated between 12 KR and 12 no-KR responses produced better retention than both the group that alternated between 6 KR and 6 no-KR responses and an all-KR group. The partial KR group that performed the best on retention also received the least number of reversals from KR to no-KR responses, however. In Experiment 2, when acquisition KR reversals ere held constant for partial KR groups, groups that received either random KR scheduling or all KR produced similar and better retention that groups who received blocked KR scheduling. These results were reconciled with KR frequency experiments by proposing that memory processes invoked by KR protocols decrease from KR frequency, to reversal, to scheduling conditions. PMID- 12529207 TI - Limb Segment Recruitment as Function of Movement Direction, Amplitude, and Speed. AB - Coordination of limb segments in graphic motor behavior has been studied primarily in cyclic tasks. In the present study, limb segment recruitment patterns were investigated in a discrete line-drawing task. Subjects (N = 11) performed pointing movements varying in direction, amplitude, and speed. The contributions of index finger, hand, and arm to the movement were analyzed by evaluating the angular displacements in 7 joint dimensions. The results showed that amplitude and direction affected limb segment involvement in the same way they have been reported to affect it in cyclic movements. Upward left- (up-left) directed movements were primarily achieved by fingers and arm, whereas upward right- (up-right) directed movements were accomplished with the hand and the arm. Large amplitudes elicited not only an increase of proximal but also a decrease of distal limb segment involvement, especially in the up-left direction. In the present discrete pointing task, effects of speed on limb segment involvement were different from speed effects that were observed earlier in cyclic tasks: Larger limb segments became more involved in fast than in slow discrete movements. With respect to the timing of limb segment recruitment, all joints tended to move simultaneously, but small deviations from synchronous joint movement onset and offset were present. The results are discussed in the context of recent theories of limb segment coordination. PMID- 12529208 TI - Diffusive, Synaptic, and Synergetic Coupling: An Evaluation Through In-Phase and Antiphase Rhythmic Movements. AB - The in-phase and antiphase patterns of interlimb l:1 frequency locking were contrasted with respect to models of coordination dynamics in biological movement systems that are based on diffusive coupling, synaptic coupling, and synergetic principles. Predictions were made from each model concerning the stable relative phase phi between the rhythmic units, its standard deviation SDphi and the self chosen coupled frequency omegasubc;. The experimental task involved human subjects oscillating two handheld pendulums either in-phase or antiphase. The eigenfrequencies of the two hand-pendulum systems were manipulated by varying the length and mass of each pendulum individually. Relative to an eigenfrequency difference of Delta equal to zero, |Deltaomega| > 0 displaced phi from phi = 0 and phi = pi, and amplified SDphi. omegasubc; decreased with |Deltaomega|. Both the displacement of phi and SDphi were greater in the antiphase mode. Additionally, the displacement of phi increased more sharply with |Delta| for antiphase than for in-phase coordination. In contrast, omegasubc; was identical for the two coordination modes. Of the models of interlimb coordination dynamics, the synergetic model was the most successful in addressing the pattern of dependencies of phi and SDphi. The specific forms of the functions relating omegasubc; and phi to Deltaomega pose challenges for all three models, however PMID- 12529209 TI - The Three-Dimensional Curvature of Straight-Ahead Movements. AB - Three-dimensional curvature of point-to-point hand movements in the forward direction was examined. Subjects (N = 4) moved their hand from a position above the start point to a forward position above targets of different size and distance. Paths were curved as a result of an initial lateral and downward movement that was compensated for in the second half of the movement. The downward component of motion had a bell-shaped velocity profile and was temporally coupled to the forward motion. Curvature was greater for movements to near targets. Examination of the relation between kinematics and geometry revealed that velocity was related to radius of curvature by a power law with an exponent of 0.59. Simulations of the component of motion in the vertical plane reproduced the qualitative behavior of curvature and fit a power law relationship between velocity and radius of curvature PMID- 12529210 TI - Afferent Information for Motor Control: The Role of Visual Information in Different Portions of the Movement. AB - The question addressed in the present study was whether subjects (N = 24) can use visual information about their hand, in the first half of an aiming movement, to ensure optimal directional accuracy of their aiming movements. Four groups of subjects practiced an aiming task in either a complete vision condition, a no vision condition, or in a condition in which their hand was visible for the first half [initial vision condition (IV)] or the second half of the movement [final vision condition (FV)]. Following 240 trials of acquisition, all subjects were submitted to a transfer test that consisted of 40 trials performed in a no-vision condition. The results indicated that seeing the hand early in movement did not help subjects to optimize either directional or amplitude accuracy. On the other hand, when subjects viewed their hand closer to the target, movements resulted that were as accurate as those performed under a complete vision condition. In transfer, withdrawing vision did not cause any increase in aiming error for the IV or the no-vision conditions. These results replicated those of Carlton (1981) and extended those of Bard and colleagues (Bard, Hay, & Fleury, 1985) in that they indicated that the kinetic visual channel hypothesized by Paillard (1980; Paillard & Amblard, 1985) appeared to be inoperative beyond 40deg of visual angle. PMID- 12529211 TI - Minimum Muscle-Tension Change Trajectories Predicted by Using a 17-Muscle Model of the Monkey's Arm. AB - Four computational problems to be solved for visually guided reaching movements, hand path, and trajectory formations, coordinate transformation, and calculations of muscle tensions are ill-posed in redundant biological control systems. These problems are ill-posed in the sense that there exist an infinite number of possible solutions. In this article, it is shown that the nervous system can solve those problems simultaneously by imposing a single global constraint: finding the smoothest muscle- tension trajectory that satisfies the desired final hand position, velocity, and acceleration. Horizontal trajectories were simulated by using a l7-muscle model of the monkey's arm as the controlled object. The simulations predicted gently curved hand paths for lateral hand movements and for movements from the side of the body to the front, and a roughly straight hand path for anterioposterior movements. The tangential hand velocities were roughly bell shaped. The simulated results were in agreement with the actual biological movements. PMID- 12529212 TI - Transitions in Visual Proprioception: A Cross-Sectional Developmental Study of the Effect of Visual Flow on Postural Control. AB - In the present study, a moving room paradigm was used that characterized the developmental progression of the effects of visual perturbations on stance control in subjects (N = 39) from 5 months to 10 years of age. Kinematic (probability of recording sway, magnitude of sway response) and electromyographic (probability and patterns of muscle activation, muscle onset latencies) data were found that suggested that visual flow simulating sway activates organized postural muscle responses and results in subsequent sway in standing infants as young as 5 months of age, well before they are able to stand independently. In new walkers, there was an increase in the magnitude of the effect of the visual perturbation, suggesting a possible increase in reliance on visual information. The magnitude of sway decreased to very low levels in older children and adults. The large-amplitude responses observed in the youngest age groups may indicate an inability to switch from an unreliable to a reliable source of perceptual information or an inability to modulate the responses produced following the perturbations. With increasing age and experience, the ability to resolve the conflict increased, with adult subjects demonstrating little sway response. PMID- 12529213 TI - The Use of Dimensional Analysis in Movement Studies. AB - Application of the technique of dimensional analysis to problems in the study of human movements is illustrated. The method is of use in design of experiments, for presentation of data, and for investigating possible relationships between dependent and independent variables. PMID- 12529214 TI - Task-dependent changes in visual feedback control: a frequency analysis of human manual tracking. AB - Prominent components in the frequency spectrum of human manual tracking responses are thought to reflect the visual feedback control loop and have been used in estimations of the visual feedback loop delay. The frequency structure of human tracking was therefore examined here in two tasks: visually guided tracking of slow and fast pseudorandom targets. Visually related frequency components were identified by testing, in each condition, the effect of adding additional feedback delays on the frequency spectrum. The major frequency components of the responses consisted of a fundamental component and its odd harmonics. These components were related to the visual feedback loop delay and shifted in concert toward lower frequencies as the feedback delay was increased. Furthermore, there were no differences in responses between 3 normal subjects and 1 subject with peripheral sensory loss. This implies that the frequency structure is dominated by the visual feedback control loop, without significant influence from proprioceptive control loops. However, the feedback-loop delay was shown to decrease from around 341 to 264 ms as the task speed doubled. Thus the estimates of visual feedback delays are influenced by the target being followed, and this suggests that the subjects can lquot;tunerquot: their feedback system to suit the demands of the tracking task. PMID- 12529215 TI - The Organization of Multisegmental Pulls Made by Standing Humans: II. Submaximal Pulls. AB - Previously, an autonomous oscillator model with three parameters was derived that describes the relationship between anterior-posterior center of mass motions and pulling force for near-maximal, bimanual pulls made by standing subjects (Michaels, Lee, Pai, 1993). The present study evaluated the extent to which a full range of pulling forces could be fit by the model and how the model's three parameters changed with intended pulling force. How much variation in force each parameter could contribute was determined by simulating the model. Qualitative and quantitative analyses of pulls made by 6 well-practiced subjects at 5prcent;, 10prcent;, 20prcent;, 40prcent;, 60prcent;, 80prcent;, and 95prcent; of their maximum pulling force revealed that the model holds well, except for the least forceful pulls of some subjects. Two parameters appeared to be controlled; one, related to the position of the center of pressure, varied most among less forceful pulls; the second, related to the position of the center of mass at the time of handle-force onset, varied most among more forceful pulls. How these parameters might be set is discussed. PMID- 12529216 TI - Practice and Component Submovements: The Roles of Programming and Feedback in Rapid Aimed Limb Movements. AB - An experiment is reported in which subjects (N = 30) practiced rapid aimed limb movements (wrist rotations), with either full visual feedback, no visual feedback, or delayed visual feedback. Previous research has indicated that, with practice, subjects increase the distance and duration of ballistic primary submovements and decrease the distance and duration of corrective secondary submovements. The design of the present experiment permitted the determination of whether these practice-related changes are results of improved programming of the primary submovement or of more efficient processing involved in the production of secondary submovements. The results suggest that improved programming occurs with practice. Furthermore, it appears that visual feedback is not solely responsible for the production of secondary submovements. PMID- 12529217 TI - Cinematographical Analysis of Movement Pathway Constraints in Rapid Target Striking Tasks. AB - Several features of the actual movement pathway in two rapid target-striking tasks were quantified by using high-speed cinematography, and whether the movement pathway is constrained as a function of the accuracy demands imposed by the size of the subtended angle was determined. Subjects (N = 16) first hit an 8 cm-diameter target located 10 cm to the left of a start position and then, depending on the condition, moved another 10 cm to hit either a 6-cm- or 1.5-cm diameter target. Subtended angles were 17.1 and 4.3 degrees for the large and small second-target conditions, respectively. Fifty trials per condition were performed, the last 3 of which were filmed at 120 Hz. The vertical dimension of movement (peak height along the z-axis) was captured directly from the camera view, whereas the horizontal (y-axis) dimension, that is, the dimension orthogonal to the principal direction of motion, was captured through a mirror positioned above the target board. Reaction times and movement times were significantly longer in the small second-target condition, thus replicating the well-known response complexity effect. Kinematic analyses revealed that when the subtended angle was smaller, there was significantly less horizontal pathway deviation as well as significantly higher peak vertical displacement in the movement. Therefore, the accuracy demands imposed by a smaller subtended angle do constrain the actual movement pathway. PMID- 12529218 TI - One-Handed Juggling: A Dynamical Approach to a Rhythmic Movement Task. AB - The skill of rhythmically juggling a ball on a racket was investigated from the viewpoint of nonlinear dynamics. The difference equations that model the dynamical system were analyzed by means of local and nonlocal stability analyses. These analyses showed that the task dynamics offer an economical juggling pattern that is stable even for open-loop actuator motion. For this pattern, two types of predictions were extracted: (a) Stable periodic bouncing is sufficiently characterized by a negative acceleration of the racket at the moment of impact with the ball, and (b) a nonlinear scaling relation maps different juggling trajectories onto one topologically equivalent dynamical system. The relevance of these results for the human control of action was evaluated in an experiment in which subjects (N = 6) performed a comparable task of juggling a ball on a paddle. Task manipulations involved different juggling heights and gravity conditions of the ball. The following predictions were confirmed: (a) For stable rhythmic performance, the paddle's acceleration at impact is negative and fluctuations of the impact acceleration follow predictions from global stability analysis; and (b) for each subject, the realizations of juggling for the different experimental conditions are related by the scaling relation. These results permit one to conclude that humans reliably exploit the stable solutions inherent to the dynamics of the given task and do not overrule these dynamics by other control mechanisms. The dynamical scaling serves as an efficient principle for generating different movement realizations from only a few parameter changes and is discussed as a dynamical formalization of the principle of motor equivalence. PMID- 12529219 TI - Resonance Tuning in Rhythmic Arm Movements. AB - The hypothesis was tested that the preferred frequency of rhythmic movement corresponds to the resonant frequency of the muscle-limb system, as proposed by the hybrid spring-pendulum model (Kugler Turvey, 1987). In contrast to previous studies, the resonant frequency and stiffness of the system were estimated independently, which permitted quantitative predictions of the preferred frequency to be made. Human subjects (N = 5) were asked to oscillate their forearms in the vertical plane at their preferred frequency under conditions of added mass and external spring loading. Subjects also oscillated their arms at frequencies below and above the preferred frequency, which enabled the investigators to estimate the resonant frequency and stiffness of the elbow joint by using the phase transfer method (Viviani, Soechting, Terzuolo, 1976). The preferred frequency corresponded to the resonant frequency of the muscle-limb system under each condition, as predicted. The oscillation amplitude varied inversely with the preferred frequency, which was also predicted. Finally, the internal joint stiffness was modulated so that it matched the impedance of the external springs but was unaffected by added mass. The results are consistent with an autonomous oscillator model that incorporates proprioception about the dynamics of the periphery. PMID- 12529220 TI - Self-Optimization of Walking in Nondisabled Children and Children With Spastic Hemiplegic Cerebral Palsy. AB - Children voluntarily adopt a frequency and movement pattern for walking. The force-driven harmonic oscillator (FDHO) model was used in this study for accurate prediction of the preferred walking frequency of nondisabled children and children with spastic hemiplegic cerebral palsy. Four potential optimality criteria with which the preferred walking pattern was forced to comply were examined: minimization of physiological costs, maximization of mechanical energy conservation, minimization of asymmetry in lower limb movements and minimization of variability of interlimb and intralimb coordination. Age and gender-matched nondisabled children (n = 6) and children with spastic hemiplegic cerebral palsy (n = 6) were tested under six frequency conditions of walking at a constant speed on a treadmill. For the nondisabled children, the results indicated that their preferred walking frequency could be accurately predicted by the FDHO model. They freely adopted a walking pattern that minimized physiological costs, asymmetry, and variability of inter- and intralimb coordination. For the children with spastic hemiplegic cerebral palsy, the prediction of preferred overground walking frequency required that the FDHO model be modified to account for muscle mass and leg length discrepancies between limbs and increased stiffness. Most of the children achieved the same optimality goals as the nondisabled when walking at the preferred frequency. However, the children were found to use different mechanisms to attain these goals: for example, a steeper increase observed in physiological cost at higher frequencies; a lowered center of gravity of the body, which allowed for angular symmetry; and greater variability of between joint coordination in the nonaffected limb and less variability in the affected limb. PMID- 12529221 TI - Influence of Destabilization on the Temporal Characteristics of lquot;Volitionalrquot; Stepping. AB - Previous work suggests that there may be fundamental differences between compensatory stepping responses evoked by postural perturbation and visually cued "volitional" stepping (e.g., gait initiation). In contrast to visual cueing, postural destabilization evokes an array of sensory inputs that are intrinsically linked to the mobilization of rapid compensatory responses. The hypothesis examined in this study was that this fundamental difference would lead to distinct changes in the temporal characteristics of the stepping response. Six healthy young adults were instructed to step quickly in response to either visual cueing or anterioposterior platform motion. Both forward and backward stepping responses were characterized, using measures of vertical ground reaction force. A stereotypical temporal patterning of the stepping response occurred in both stimulus conditions and both directions of stepping, and anticipatory postural adjustments were evident in all trials. However, postural destabilization led to a more rapid initiation and execution of the temporal pattern, in comparison with visually cued responses. The most pronounced effect was seen in the duration of the response, which was reduced by a factor of two, with approximately proportional foreshortening of both the preparatory and swing phases. The results suggest that sensory information conveying a state of instability has a distinct influence on the characteristics of a subsequent stepping reaction. The persistence of the anticipatory postural adjustments suggests that the failure to see this element of the response in previous studies may reflect fundamental differences between volitional and unplanned compensatory stepping. PMID- 12529222 TI - Locomotor Patterns of the Leading and the Trailing Limbs as Solid and Fragile Obstacles Are Stepped Over: Some Insights Into the Role of Vision During Locomotion. AB - The issues explored in this article are the role of exproprioceptive input and the nature of exteroceptive input provided by the visual system in the control of limb elevation as obstacles are stepped over during locomotion. In the first experiment, the differences in limb trajectory of movements over solid and fragile obstacles of similar dimensions were examined. Subjects increased their toe clearance, vertical position of the hip, and the hip vertical velocity when going over a fragile obstacle with the leading limb. This suggests that in addition to visually observable properties of obstacles such as height or width, other properties, such as rigidity or fragility, which may be classified as visually inferred, also influence the limb trajectory. Part of the first and the second experiment was focused on understanding differences in leading limb and trailing limb trajectory over obstacles. The toe clearance of the trailing limb was lower for smaller obstacles. There was no consistent correlation between the toe clearance values of the leading and trailing limbs. The variability in toe clearance was higher for the trailing limb, which is attributable to lack of visual exproprioceptive input about trailing limb movements and to the shorter time available following toe-off to fine-tune the trailing limb trajectory. Because the body center of mass is moving toward the supporting foot when the trailing limb goes over obstacles and the trailing limb foot is moving up, the chances of a trip are minimized and recovery from an unexpected trip are more likely. These results highlight the role of exproprioceptive input provided by the visual system and possible cognitive influences on the limb trajectory as one travels over uneven terrains. PMID- 12529223 TI - Coordination Between Breathing and Finger Tracking in Man. AB - Arm and leg movements are known to produce temporal pattern changes of breathing. This can be interpreted as coordination, as defined by von Holst (1939). The aim of the present study was to find whether breathing exerts an influence in a reverse direction on a nonrespiratory movement as well. A pursuit tracking test was used, and test individuals (N = 19) were instructed to track a visually presented step function by flexion or extension of their right index finger. Velocity and precision of the step responses proved to be dependent on their relation to the breathing time course; the differences between inspiratory and expiratory responses were smaller than those within each half-cycle. The movements were performed more rapidly and more precisely in about the middle of each half-cycle than immediately after the respiratory phase transition or during the second half of each inspiration or expiration. Discontinuous short-lasting motor actions exerted a coordinative influence on respiration comparablewith that of periodical events: Breaths coinciding with step responses were shortened, preferably when the preset step was given early in the inspiration. It was hypothesized that the reciprocal effect between both motor actions changes periodically. In the first part of each respiratory half-cycle, the respiratory rhythm exerts only a weak influence on additional movements, but it can be altered easily by simultaneous motor processes. Toward the respiratory phase switching, the respiratory rhythm behaves more stably against coordinative influences and becomes capable of impairing an additional movement. PMID- 12529224 TI - Longitudinal Development of the Automatic Postural Response in Infants. AB - In this longitudinal study, the development of automatic postural responses elicited during stance following perturbation of the support surface was investigated. Infants (N = 9) unable to stand without support were tested initially: follow-up tests were performed until the infants were able to stand and walk independently. Surface electromyographic recordings of leg and trunk muscle activity following a postural perturbation induced by a forward or backward translation of the support surface were made for each infant. Muscle onset latencies following the perturbation and the proportion of trials in which muscle activity was recorded were determined. First, infants activated appropriate muscles either in isolation or in pairs and then combined these muscles into functional synergies. Although activation of all three postural muscles was recorded in infants before they were able to stand and walk independently, the three-muscle response was not consistent in the youngest children. The proportion of trials eliciting muscle activity continued to increase (p <.O5) after infants began to walk independently, with postural muscle activity recorded in virtually every trial by late independent walking. Thus. the automatic postural response elicited during stance was shown to begin with activity in single or paired muscles, followed by activation of the postural muscles in functional synergies. These data illustrate the progressive development of an effective sensory-motor organization. PMID- 12529225 TI - Practicing a Structured Continuous Key-Pressing Task: Motor Chunking or Rhythm Consolidation? AB - An experiment is reported on the effects of extensive practice in a task in which subjects (N = 36) pressed a succession of nine keys with nine separate fingers, each key press in response to a corresponding stimulus. The order of the key presses remained constant over practice. Key-pressing cycles followed each other without interruption. A stimulus was usually presented immediately upon depressing the previous key, but in the structured conditions a stimulus was preceded at either two (the 45 group) or three (the 333 group) positions by a 750 ms response-stimulus interval (RSI). This partitioned the sequence into three response groups for subjects in the 333 condition and into two response groups for subjects in the 45 condition. On occasion, all subjects performed in the unstructured condition, in which RSls were zero. Interkey times in this condition clearly reflected the position of the 750-ms RSls in the structured conditions. This suggests that motor chunks developed in the structured conditions were also used in the unstructured condition. Rhythm-based control, as proposed by Summers (1975), was rejected as an explanation for this effect because group-start-within group ratios exceeded 2:1 and because the theory of rhythm-based control could not predict intervals in the unstructured 45 condition. Unstructured within-group intervals were slower than structured within-group intervals, which effect was more pronounced in the 333 than in the 45 condition. Also, the initial element of the unstructured four-key group was faster than the initial element of the unstructured three-key group. These and other findings (a) accord with the notion that in unstructured sequences preparing a forthcoming response group concurs with the execution of the preceding group and (b) suggest that shorter groups are more slowed by concurrent preparation for the forthcoming group than are longer groups. PMID- 12529226 TI - Variability of Practice and Contextual Interference in Motor Skill Learning. AB - The purpose of this study was to investigate whether learning benefits in multiple-task learning situations are a result of contextual interference or of schema enhancement related to the amount of variability in the practice session. Two experiments were designed that replicated and extended the experiment reported by Wulf and Schmidt (1988). In a 2 (same vs. different relative time) x 2 (blocked vs. random practice schedule) design, 48 right-handed subjects were randomly assigned to one of four experimental conditions. A tapping task was employed that required a right-handed tap of three small brass plates arranged in a diamond pattern. Each segment had a specific time requirement. Target times and response times were provided on a computer screen directly in front of the subject. Each subject participated in two acquisition sessions (i.e., 198 practice trials) and was tested for learning on several different retention and transfer tests. In Experiment 2, a control group was added that received no acquisition phase. Results of both experiments showed a typical contextual interference effect, with depressed scores by the random groups during acquisition but significantly better scores than the blocked groups on several retention and transfer tests. Certain characteristics of the tests were found to influence the demonstration of the practice schedule effects. These results were consistent with predictions from Magill and Hall (1990) that the learning benefits of contextual interference are more likely to occur when skill variations are from different classes of movement and that the amount of variability in practice is more influential when the to-be-learned tasks are parameter modifications of the same generalized motor program. PMID- 12529227 TI - Evidence of Incomplete Motor Programming in Parkinson's Disease. AB - One of the essential questions regarding movement deficits in Parkinson's disease (PD) is whether they stem from impaired selecting and switching among movements, impaired use of predictive information to prepare movement, or impaired execution of movement. PD subjects (n equals 9) and age-matched control subjects (n equals 8) performed a cued, sequential-response RT task. The cue provided either no information, accurate information, or inaccurate information about the upcoming response. PD subjects used predictive information to prepare and to switch among movement sequences normally, but second and third key press latencies were prolonged in comparison with the first key press latency. In Experiments 2 and 3, the effects of choice set and sequence length on key press latencies were examined. These results provide evidence that PD subjects initiate movement before the entire response sequence is prepared. PD does not impair motor programming or execution processes themselves but impairs the smooth coordination of those processes. PMID- 12529228 TI - Investigating Individual Differences in a Serial Choice Reaction Time Task: Use of Auditory Feedback and Analysis of Responses Surrounding Errors. AB - Subjects (N = 78) performed a visual four-choice reaction time (RT) task, either with or without immediate trial-by-trial feedback, in which RT (but not accuracy) was indicated by the pitch of an auditory tone. For each feedback condition, half of the subjects (the high AH4 group) scored more than 50% on the AH4 test of fluid intelligence (Heim, 1968), whereas the remaining half (the low AH4 group) scored less than 50%. It was predicted that if low AH4 subjects were slow because they were poor at monitoring RT, they would benefit more from feedback than high AH4 subjects would. This was not supported by the data: There was some beneficial effect of feedback on RT, but only for the high AH4 group. A second possibility was that individual differences would be apparent in processes such as detecting errors and controlling RT from trial to trial. From analyses of error rates, RT distributions, and particularly sequences of responses before and after errors, there was no evidence of qualitative differences in performance between the high and low AH4 groups. It is concluded that individual differences in this task are largely determined by information-processing rate rather than by factors such as the ability to detect errors or to monitor and control RT. PMID- 12529229 TI - Achieving Coordination in Prehension: Joint Freezing and Postural Contributions. AB - The focus of the present study was on the intersegmental relationships that emerge when both task and oganismic constraints are imposed upon the coordination system. Seven right-handed subjects were required to reach and grasp a cup (hand transport phase) and place it on a designated target (cup transport phase), using either their preferred or nonpreferred hand. The kinematics of the movement were examined as a function of task (grasping a full cup versus grasping an empty one) and organismic (preferred or nonpreferred hand) constraints. During the hand transport phase, a task constraint effect was revealed through an increase in the low-velocity phase for the full cup condition. This constraint coexisted with a decrease in angular motion of the shoulder and elbow joints, indicating subjects reduced the number of variables to be independently controlled in the final homing-in stage of the movement. Accompanying this decrease in angular change was an increase in the displacement of the trunk. During the cup transport phase, the trunk was shown to contribute significantly more to the movement in the full cup condition and for the left hand movements, thereby increasing the stability of the movement system. These findings are in agreement with Bernstein's (1967) notion of fixating parts of the body as an initial solution to a movement problem, and they lend support to the concept of a proximodistal organization of coordination. PMID- 12529230 TI - Amplitude Scaling Compensates for Serial Delays in Correcting Eye and Arm Movements. AB - Spatial and metrical parameters of the eye and arm movements made by human subjects (N = 7) in response to visual targets that were stepped unexpectedly either once (single step) or twice (double step) were studied. For the double step, the displacement of a visual target was decreased or increased in amplitude at intervals before and during a movement. Provided the second target step occurred more than 100 ms before the onset of movement, the amplitude of the subjects' first response was altered in the direction of the new target location. But this amplitude scaling was not always sufficient to reach the new target location, and a second corrective response was required. The latency in producing this second response was greatly increased above reaction time latencies of movements to single-step targets, especially when the target change occurred 100 ms or more before movement onset. These findings suggest that even though serial processing limitations delay the production of a second corrective response, continuous parallel processing of visual information enables the amplitude of the first response to be altered with minimal delay. This enables some degree of real time continuous control by the visuomotor control system. PMID- 12529231 TI - Adaptive Dynamics of the Leg Movement Patterns of Human Infants: III. Age-Related Differences in Limb Control. AB - In this article, the development of the increasingly differentiated control of the joints necessary to transform the spontaneous leg movements of early infancy into adaptive and functional actions is described. The hypothesis-that increasing joint control requires the capability for disassociation of joint action, the active modulation of joint stiffness, and a transition from proximal to distal control of the joints-is proposed. Kinematic and kinetic analyses of the vertical kicks of infants 2 weeks, 3 months, and 7 months of age (as well as a comparative group of adults) indicated increasing joint independence as well as phase dependent and joint-dependent control modifications. The kicks of the younger infants were dominated by a proximal control strategy and minimal adjustments of the limb energetics during the flexion and extension phases of the kick. By 7 months of age, much larger modulations of the kick phases were observed as well as increasing evidence of distal control. These results revealed kinematic and kinetic patterns of emerging limb control between 2 weeks and 7 months of age. PMID- 12529232 TI - Variation of Isometric Response Force in the Rat. AB - Hungry, unrestrained rats (N = 7) were rewarded for pressing a response beam in excess of 11 different force requirements. Changes in peak force production as a function of peak force requirement were examined by analyses of the first four moments of distributions of peak response forces: constant error, the within subject standard deviation, skewness, and kurtosis. Results were similar to those previously obtained with human subjects: Constant error was positive at low and negative at high force requirements, the within-subject standard deviation increased as a negatively accelerating function of force requirement, and skewness and kurtosis were positive at low force requirements and decreased to negative values at the highest increments. Additional analyses of response kinetics indicated that rats, like humans, meet increasing force requirements by altering the rate of rise of force. The performance similarities suggest that common processes are engaged by the human and rat motor control systems to solve the problem of generating forces that are appropriate to the prevailing environmental constraints. PMID- 12529233 TI - Phase Transitions and Critical Fluctuations in Rhythmic Coordination of Ipsilateral Hand and Foot. AB - Four subjects performed rhythmic movements of the ankle and the wrist in time with an auditory metronome, in two modes of coordination, antiphase and in-phase. The forearm was placed in either a prone or a supine position. When movements were prepared in the antiphase mode, spontaneous transitions to the in-phase mode, or to phase wandering were observed as metronome frequency was increased. When prepared in the in-phase mode, transitions between in-phase modes or to phase wandering were occasionally observed. Predicted signature features of nonequilbrium phase transitions were noted, including loss of stability and critical fluctuations. The stability of the movement patterns was determined by spatial (dependent upon the direction of movement) rather than anatomical (dependent on the coupling of specific muscle groups) constraints. The position of the forearm had no consistent bearing upon the variability of the phase relations between the limbs, the frequency of phase transitions, or the time of onset of transitions. These results are discussed with reference to the coordination dynamics (e.g., multistability, loss of stability) of multijoint movements. PMID- 12529234 TI - Controlling the Ground Reaction Force During Lifting. AB - The control of the ground reaction force vector relative to the center of gravity (CoG) was examined while subjects performed a back-lifting task. Six male subjects (aged 24.0 +/- 2.5 years) repeatedly lifted a barbell. A biomechanical analysis that used a linked segment model revealed that the summed rotations of body segments during lifting yielded a specific rate of change of the angular momentum of the entire body. This equaled the external moment provided by Fsubg; relative to CoG. This implies that multisegment movements involve control of the angular momentum of the entire body through an appropriately directed Fsubg;. Thus, in dynamic tasks Fsubg; is pointed away from rather than lined up with the CoG, as is the case in static tasks. PMID- 12529235 TI - On the Problem of Two-Dimensional Error Scores: Measures and Analyses of Accuracy, Bias, and Consistency. AB - Describing and analyzing error for one-dimensional performance tasks is fairly straightforward, but suggestions for describing and analyzing error for two dimensional performance tasks (e.g., marksmanship) are quite problematic. Specifically, imposing an arbitrary axis onto the two-dimensional work space, along which traditional one-dimensional measures can be computed and analyzed, yields measures of accuracy, bias, and consistency that are entirely dependent upon the choice of axis. The present contribution offers new measures and methods for describing and analyzing data from two-dimensional performances. Unlike the resu1ts from previous suggestions, the approaches described herein yield results that are completely independent of the axes used to quantify the individual two dimensional trials. These new approaches are strongly related to well-established methods for describing and analyzing error for one-dimensional tasks. PMID- 12529236 TI - Finger Pairings in Two-Choice Reaction Time Taskscolon: Does the Between-Hands Advantage Reflect Response Preparation? AB - Two fractionated RT experiments tested whether the response-preparation or response-implementation hypothesis better accounts for the observation that two choice reaction time (RT) usually takes longer when the responses are performed by the fingers of the same hand (within-hand repertoire) than by the fingers of the two hands (between-hands repertoire). In Experiment I (n equals 8), the effect of repertoire on the premotor time and the motor time were studied. RT was divided into the two periods with respect to the onset of change in electromyographic (EMG) activity of the flexor digitorum profundus. Type of repertoire affected both time periods. In Experiment 2 (n = 16), the effects of repertoire and foreperiod duration on the premotor and motor times of the flexor digitorum profundus and flexor digitorum sublimis were studied. The results of Experiment I were confirmed, and the effects of repertoire and foreperiod duration were found to be additive on premotor time but interactive on motor time. These findings led to rejection of the response-preparation hypothesis and instead supported the view that the central command for the flexion of the right middle finger differs according to the type of repertoire. The command appears to specify a lower rate of recruitment of the prime movers in the within-hand repertoire than in the between-hands repertoire. The execution of the central commands may depend on the state of excitability of the spinal neurons. Analysis of the EMG signals revealed that speed of contraction of the prime movers depends on repertoire when the foreperiod is long but not when it is short. The additivity of the effects of repertoire and of foreperiod duration on premotor time support the view that regardless of the state of preparation of the subject the pattern of EMG activity required for flexion of the right middle finger in each repertoire is specified during the premotor time. PMID- 12529237 TI - Relative Phase Alterations During Bimanual Skill Acquisition. AB - The purposes of the research reported here were (a) to examine changes in relative phase during the acquisition of a new coordination pattern and (b) to determine the effect of learning this pattern on the ability to perform other coordination patterns. Ten subjects practiced an upper limb coordination task that required a 90 degrees phase offset and different amplitudes for each arm. A gross approximation of the mean relative phase for the intended coordination pattern occurred quickly, but the attainment of stability occurred much more gradually. These results were accompanied by changes in pattern stability across practice and on various transfer tests. Learning of the new coordination pattern also affected the stability of the antiphase mode, but this effect was only temporary. PMID- 12529238 TI - A Forthcoming Key Press Can Be Selected While Earlier Ones Are Executed. AB - The possibility that a key press can be selected during execution of earlier key presses and the resulting pattern of interferences were investigated in this study. Subjects (N equals 26) were required to press a series of keys, determined in advance, before they pressed a stimulus-dependent key. Response selection demands were manipulated by using spatially compatible and incompatible SndashR mappings because S-R compatibility is well known to not disappear with practice. The longer time needed to select an incompatible key vanished when the choice key was preceded by two or four predetermined key presses. Only early in practice did the time to press the first and the choice key in a three-key sequence exhibit part of the compatibility effect. With limited practice, concurrent preparation in the three-key sequence was relatively slow and took longer than the time required for executing the fixed keys. These findings suggest that processes involved in execution are not affected by concurrent response selection and that one of the effects of practicing movement sequences is that later movements can be selected while earlier ones are being executed. This need not affect execution rate. Therefore, different degrees of concurrent processing are not always reflected in reduced execution rates. PMID- 12529239 TI - A Method for Identifying the Initiation of Reaching Movements in Natural Prehension. AB - Identifying the start of a movement is critical for calculating variables such as movement duration, movement speed, or trajectory length and straightness. In most experiments, the problem is simplified by giving instructions to the subjects and by constraining the task. This is not possible in populations that are too young or disabled to understand and execute the task as instructed. In this article, a method is presented for reliably identifying the onset of reaches in infants, whose movements are embedded in their ongoing activities. A computer program prompts the user to choose reach initiation on the basis of comparison between hand kinematics and videotaped behavior and to code the reach as emerging from 1 of 4 movement contexts. The method is highly reliable in infants of all ages and captures expected age-related changes in reaching. It is suggested that researchers could use this or a similar method to study movements that are part of natural, ongoing activities in normal and in clinical populations PMID- 12529240 TI - Depolarization-induced slow calcium transients activate early genes in skeletal muscle cells. AB - The signaling mechanisms by which skeletal muscle electrical activity leads to changes in gene expression remain largely undefined. We have reported that myotube depolarization induces calcium signals in the cytosol and nucleus via inositol 1,4,5-trisphosphate (IP(3)) and phosphorylation of both ERK1/2 and cAMP response element-binding protein (CREB). We now describe the calcium dependence of P-CREB and P-ERK induction and of the increases in mRNA of the early genes c fos, c-jun, and egr-1. Increased phosphorylation and early gene activation were maintained in the absence of extracellular calcium, while the increase in intracellular calcium induced by caffeine could mimic the depolarization stimulus. Depolarization performed either in the presence of the IP(3) inhibitors 2-aminoethoxydiphenyl borate or xestospongin C or on cells loaded with BAPTA-AM, in which slow calcium signals were abolished, resulted in decreased activation of the early genes examined. Both early gene activation and CREB phosphorylation were inhibited by ERK phosphorylation blockade. These data suggest a role for calcium in the transcription-related events that follow membrane depolarization in muscle cells. PMID- 12529241 TI - Dual effect of nitric oxide on cytosolic Ca2+ concentration and insulin secretion in rat pancreatic beta-cells. AB - In isolated rat pancreatic beta-cells, the nitric oxide (NO) donor NOC-7 at 1 microM reduced the amplitude of the oscillations of cytosolic Ca(2+) concentration ([Ca(2+)](c)) induced by 11.1 mM glucose, and at 10 microM terminated them. In the presence of N(G)-nitro-l-arginine (l-NNA), however, NOC-7 at 0.5 and 1 microM increased the amplitude of the [Ca(2+)](c) oscillations, although the NO donor at 10 microM still suppressed them. Aqueous NO solution also had a dual effect on the [Ca(2+)](c) oscillations. The soluble guanylate cyclase inhibitor LY-83583 and the cGMP-dependent protein kinase inhibitor KT5823 inhibited the stimulatory effect of NO, and 8-bromo-cGMP increased the amplitude of the [Ca(2+)](c) oscillations. Patch-clamp analyses in the perforated configuration showed that 8-bromo-cGMP inhibited whole cell ATP-sensitive K(+) currents in the isolated rat pancreatic beta-cells, suggesting that the inhibition by cGMP of ATP-sensitive K(+) channels is, at least in part, responsible for the stimulatory effect of NO on the [Ca(2+)](c) oscillations. In the presence of l-NNA, the glucose-induced insulin secretion from isolated islets was facilitated by 0.5 microM NOC-7, whereas it was suppressed by 10 microM NOC 7. These results suggest that NO facilitates glucose-induced [Ca(2+)](c) oscillations of beta-cells and insulin secretion at low concentrations, which effects are mediated by cGMP, whereas NO inhibits them in a cGMP-independent manner at high concentrations. PMID- 12529242 TI - Glutamine delays spontaneous apoptosis in neutrophils. AB - Nuclear, mitochondrial, and plasma membrane events associated with apoptosis were investigated in rat neutrophils cultivated for 3, 24, and 48 h in the absence or presence of glutamine (0.5, 1.0, and 2.0 mM). Condensation of chromatin was reduced after 24 or 48 h of culture in the presence of glutamine compared with its absence as assessed by Hoechst 33342 staining. The level of Escherichia coli phagocytosis in the presence of glutamine was markedly increased compared with the level achieved by cells cultured in the absence of glutamine. Annexin V binding to externalized phosphatidylserine was reduced in the presence of glutamine. Sensitive fluorochrome rhodamine 123, as determined by fluorescence activated cell sorting and confocal microscopy, was used to monitor loss of the mitochondrial transmembrane potential. In the absence of glutamine, neutrophils exhibited a marked reduction in the uptake of rhodamine 123. In the presence of 1.0 or 2.0 mM glutamine, the uptake of rhodamine was 20 or 38% higher, respectively. Similar effect was found in human neutrophils by measuring DNA fragmentation and mitochondrial transmembrane potential. Therefore, glutamine protects from events associated with triggering and executing apoptosis in both rat and human neutrophils. PMID- 12529243 TI - Exogenous eosinophil activation converts PSGL-1-dependent binding to CD18 dependent stable adhesion to platelets in shear flow. AB - This study examined the binding kinetics and molecular requirements of eosinophil adhesion to surface-anchored platelets in shear flow. P-selectin glycoprotein ligand-1 (PSGL-1) binding to platelet P-selectin initiates tethering and rolling of eosinophils to platelets under flow. These primary interacting cells assist in the capture of free-flowing eosinophils through homotypic tethering (secondary interactions) mediated via L-selectin-PSGL-1 interactions. Differences between eosinophils and neutrophils in PSGL-1 and L-selectin expression levels predict the pattern and relative extent of their adhesive interactions with immobilized platelets under shear, as well as the relative magnitude of their average rolling velocities. The majority of tethered eosinophils become rapidly stationary on the platelet layer, a process that is predominantly mediated via eosinophil PSGL-1 binding to platelet P-selectin and has an absolute requirement for intact cytoskeleton. Only a small fraction of these stationary eosinophils develop shear resistant attachments mediated by CD18 integrins. However, stimulation of eosinophils with eotaxin-2 converts PSGL-1-P-selectin-dependent stationary adhesion to CD18-mediated shear-resistant stable attachment. These studies provide insights for designing strategies based on blocking of eosinophil platelet interactions to combat thrombotic disorders in hypereosinophilic patients. PMID- 12529244 TI - Transcriptional regulation of the human NaPi-IIb cotransporter by EGF in Caco-2 cells involves c-myb. AB - The type IIb sodium-phosphate (NaP(i)-IIb) cotransporter mediates intestinal phosphate absorption. Previous work in our laboratory has shown that EGF inhibited NaP(i)-IIb cotransporter expression through transcriptional regulation. To understand this regulation, progressively shorter human NaP(i)-IIb promoter constructs were used to define the EGF response region, and gel mobility shift assays (GMSAs) were used to characterize DNA-protein interactions. Promoter analysis determined that the EGF response region was located between -784 and 729 base pair (bp) of the promoter. GMSAs and overexpression studies revealed an interaction between this promoter region and c-myb transcription factor. Inhibition of EGF receptor activation restored promoter function. Further studies suggested that MAPK, PKC, and/or PKA pathways are involved in this regulation. In conclusion, these studies suggest that EGF decreases human NaP(i)-IIb gene expression by modifying the c-myb protein such that it inhibits transcriptional activation. We further conclude that this downregulation of promoter function is mediated by EGF-activated PKC/PKA and MAPK pathways. This is the first study that demonstrates involvement of c-myb in the regulation of intestinal nutrient absorption. PMID- 12529245 TI - The nonclassic secretion of thioredoxin is not sensitive to redox state. AB - Thioredoxin (Trx) is a cytosolic, redox-active protein that is secreted from many cells and has several extracellular functions. In activated lymphocytes, the pathway of secretion does not involve the Golgi apparatus. Levels of extracellular Trx are decreased by the antioxidant N-acetylcysteine. Hence, the secretion of Trx could be altered by the redox status of the cell or the protein. To study Trx mutants, we characterized the secretion of human Trx from Chinese hamster ovary cells. Secretion of human Trx is unaffected by brefeldin A, slow but efficient, and sensitive to low temperature and factors in serum. We demonstrate that N-acetylcysteine reduces the cellular level of Trx but not the proportion secreted; thus this chemical does not block the nonclassic pathway for Trx secretion. Furthermore, we find that mutations in either the active site or the dimerization site of Trx do not alter its secretion. Thus the nonclassic secretion of Trx is not dependent on the redox status of either the cell or the protein. PMID- 12529246 TI - Structure-function relationships of AE2 regulation by Ca(i)(2+)-sensitive stimulators NH(4+) and hypertonicity. AB - We showed previously that the nonerythroid anion exchanger AE2 and the erythroid anion exchanger AE1 differ greatly in their regulation by acute changes in intracellular pH (pH(i)) and extracellular pH (pH(o)). We have now examined how AE2, but not AE1, is activated by two stimuli with opposing effects on oocyte pH(i): an alkalinizing stimulus, hypertonicity, and an acidifying stimulus, NH(4)(+). We find that both NH(2)-terminal cytoplasmic and COOH-terminal transmembrane domains of AE2 are required for activation by either stimulus. Directed by initial deletion mutagenesis studies of the NH(2)-terminal cytoplasmic domain, an alanine scan of AE2 amino acids 336-347 identified residues whose individual mutation abolished or severely attenuated sensitivity to both or only one activating stimulus. Chelation of cytoplasmic Ca(2+) (Ca(i)(2+)) diminished or abolished AE2 stimulation by NH(4)(+) and by hypertonicity. Calmidazolium inhibited AE2 activity, but not that of AE1. AE2 was insensitive to many other modifiers of Ca(2+) signaling. Unlike AE2 stimulation by NH(4)(+) and by hypertonicity, AE2 inhibition by calmidazolium required only AE2's COOH-terminal transmembrane domain. PMID- 12529247 TI - ET-1- and NO-mediated signal transduction pathway in human brain capillary endothelial cells. AB - Previous studies have demonstrated that functional interaction between endothelin (ET)-1 and nitric oxide (NO) involves changes in Ca(2+) mobilization and cytoskeleton in human brain microvascular endothelial cells. The focus of this investigation was to examine the possible existence of analogous interplay between these vasoactive substances and elucidate their signal transduction pathways in human brain capillary endothelial cells. The results indicate that ET 1-stimulated Ca(2+) mobilization in these cells is dose-dependently inhibited by NOR-1 (an NO donor). This inhibition was prevented by ODQ (an inhibitor of guanylyl cyclase) or Rp-8-CPT-cGMPS (an inhibitor of protein kinase G). Treatment of endothelial cells with 8-bromo-cGMP reduced ET-1-induced Ca(2+) mobilization in a manner similar to that observed with NOR-1 treatment. In addition, NOR-1 or cGMP reduced Ca(2+) mobilization induced by mastoparan (an activator of G protein), inositol 1,4,5-trisphosphate, or thapsigargin (an inhibitor of Ca(2+) ATPase). Interestingly, alterations in endothelial cytoskeleton (actin and vimentin) were associated with these effects. The data indicate for the first time that the cGMP-dependent protein kinase colocalizes with actin. These changes were accompanied by altered levels of phosphorylated vasodilator-stimulated phosphoprotein, which were elevated in endothelial cells incubated with NOR-1 and significantly reduced by ODQ or Rp-8-CPT-cGMPS. The findings indicate a potential mechanism by which the functional interrelationship between ET-1 and NO plays a role in regulating capillary tone, microcirculation, and blood-brain barrier function. PMID- 12529248 TI - Role of calcium in metabolic signaling between cardiac sarcoplasmic reticulum and mitochondria in vitro. AB - The role of Ca(2+) as a cytosolic signaling molecule between porcine cardiac sarcoplasmic reticulum (SR) ATPase and mitochondrial ATP production was evaluated in vitro. The Ca(2+) sensitivity of these processes was determined individually and in a reconstituted system with SR and mitochondria in a 0.5:1 protein-to cytochrome aa(3) ratio. The half-maximal concentration (K(1/2)) of SR ATPase was 335 nM Ca(2+). The ATP synthesis dependence was similar with a K(1/2) of 243 nM for dehydrogenases and 114 nM for overall ATP production. In the reconstituted system, Ca(2+) increased thapsigargin-sensitive ATP production (maximum approximately 5-fold) with minimal changes in mitochondrial reduced nicotinamide adenine dinucleotide (NADH). NADH concentration remained stable despite graded increases in NADH turnover induced over a wide range of Ca(2+) concentrations (0 to approximately 500 nM). These data are consistent with a balanced activation of SR ATPase and mitochondrial ATP synthesis by Ca(2+) that contributes to a homeostasis of energy metabolism metabolites. It is suggested that this balanced activation by cytosolic Ca(2+) is partially responsible for the minimal alteration in energy metabolism intermediates that occurs with changes in cardiac workload in vivo. PMID- 12529249 TI - Effects of peroxynitrite on sarcoplasmic reticulum Ca2+ pump in pig coronary artery smooth muscle. AB - Peroxynitrite generated in arteries from superoxide and NO may damage Ca(2+) pumps. Here, we report the effects of peroxynitrite on ATP-dependent azide insensitive uptake of Ca(2+) into pig coronary artery vesicular membrane fractions F2 [enriched in plasma membrane (PM)] and F3 [enriched in sarcoplasmic reticulum (SR)]. Membranes were pretreated with peroxynitrite and then with DTT to quench this agent. This pretreatment inhibited Ca(2+) uptake in a peroxynitrite concentration-dependent manner, but the effect was more severe in F3 than in F2. The inhibition was thus not overcome by excess DTT used to quench peroxynitrite and was not affected if catalase, SOD, or mannitol was added along with peroxynitrite. Such damage to the pump protein would be difficult to repair if produced during ischemia-reperfusion. The acylphosphates formed with ATP in F3 corresponded mainly to the SR Ca(2+) pump (110 kDa), but in F2 both PM (140 kDa) and 110-kDa bands were observed. Peroxynitrite treatment of F2 inhibited only the 110-kDa band. Inhibition of Ca(2+) uptake and acylphosphate formation from ATP correlated well in peroxynitrite-treated F3 samples. However, inhibition of acylphosphates from orthophosphate (reverse reaction of the pump) was slightly poorer. Peroxynitrite treatment also covalently cross-linked the pump protein, yielding no dimers but only larger oligomers. In contrast, cross-linking of the SR Ca(2+) pump in skeletal and cardiac muscles gives dimers as the first oligomers. Therefore, we speculate that SERCA2 has a different quaternary structure in the coronary artery smooth muscle. PMID- 12529250 TI - PDGF stimulates pulmonary vascular smooth muscle cell proliferation by upregulating TRPC6 expression. AB - Capacitative Ca(2+) entry (CCE) through store-operated Ca(2+) (SOC) channels plays an important role in returning Ca(2+) to the sarcoplasmic reticulum (SR) and regulating cytosolic free Ca(2+) concentration ([Ca(2+)](cyt)). A rise in [Ca(2+)](cyt) and sufficient Ca(2+) in the SR are required for pulmonary artery smooth muscle cell (PASMC) proliferation. We tested the hypothesis that platelet derived growth factor (PDGF)-mediated PASMC growth involves upregulation of c-Jun and TRPC6, a transient receptor potential cation channel. In rat PASMC, PDGF (10 ng/ml for 0.5-48 h) phosphorylated signal transducer and activator of transcription (STAT3), increased mRNA and protein levels of c-Jun, and stimulated cell proliferation. PDGF treatment also upregulated TRPC6 expression and augmented CCE, elicited by passive depletion of Ca(2+) from the SR using cyclopiazonic acid. Furthermore, overexpression of c-Jun stimulated TRPC6 expression and CCE amplitude in PASMC. Downregulation of TRPC6 using an antisense oligonucleotide specifically for human TRPC6 decreased CCE and inhibited PDGF mediated PASMC proliferation. These results suggest that PDGF-mediated PASMC proliferation is associated with c-Jun/STAT3-induced upregulation of TRPC6 expression. The resultant increase in CCE raises [Ca(2+)](cyt), facilitates return of Ca(2+) to the SR, and enhances PASMC growth. PMID- 12529251 TI - cAMP-dependent exocytosis and vesicle traffic regulate CFTR and fluid transport in rat jejunum in vivo. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) channel is regulated by cAMP-dependent vesicle traffic and exocytosis to the apical membrane in some cell types, but this has not been demonstrated in the intestinal crypt. The distribution of CFTR, lactase (control), and fluid secretion were determined in rat jejunum after cAMP activation in the presence of nocodazole and primaquine to disrupt vesicle traffic. CFTR and lactase were localized by immunofluorescence, and surface proteins were detected by biotinylation of enterocytes. Immunoprecipitates from biotinylated and nonbiotinylated cells were analyzed by streptavidin detection and immunoblots. Immunolocalization confirmed a cAMP-dependent shift of CFTR but not lactase from a subapical compartment to the apical surface associated with fluid secretion that was reduced in the presence of primaquine and nocodazole. Analysis of immunoblots from immunoprecipitates after biotinylation revealed a 3.8 +/- 1.7-fold (P < 0.005) increase of surface-exposed CFTR after vasoactive intestinal peptide (VIP). These measurements provide independent corroboration supporting a role for vesicle traffic in regulating CFTR and cAMP-induced fluid transport in the intestine. PMID- 12529252 TI - The gamma-subunit of ENaC is more important for channel surface expression than the beta-subunit. AB - The amiloride-sensitive epithelial sodium channel (ENaC) plays a critical role in fluid and electrolyte homeostasis and is composed of three homologous subunits: alpha, beta, and gamma. Only heteromultimeric channels made of alphabetagammaENaC are efficiently expressed at the cell surface, resulting in maximally amiloride sensitive currents. To study the relative importance of various regions of the beta- and gamma-subunits for the expression of functional ENaC channels at the cell surface, we constructed hemagglutinin (HA)-tagged beta-gamma-chimeric subunits composed of beta- and gamma-subunit regions and coexpressed them with HA tagged alphabeta- and alphagamma-subunits in Xenopus laevis oocytes. The whole cell amiloride-sensitive sodium current (DeltaI(ami)) and surface expression of channels were assessed in parallel using the two-electrode voltage-clamp technique and a chemiluminescence assay. Because coexpression of alphagammaENaC resulted in larger DeltaI(ami) and surface expression compared with coexpression of alphabetaENaC, we hypothesized that the gamma-subunit is more important for ENaC trafficking than the beta-subunit. Using chimeras, we demonstrated that channel activity is largely preserved when the highly conserved second cysteine rich domains (CRD2) of the beta- and gamma-subunits are exchanged. In contrast, exchanging the whole extracellular loops of the beta- and the gamma-subunits largely reduced ENaC currents and ENaC expression in the membrane. This indicates that there is limited interchangeability between molecular regions of the two subunits. Interestingly, our chimera studies demonstrated that the intracellular termini and the two transmembrane domains of gammaENaC are more important for the expression of functional channels at the cell surface than the corresponding regions of betaENaC. PMID- 12529253 TI - Role of phosphoinositide 3-kinase in the nonselective cation channel activation by endothelin-1/endothelinB receptor. AB - We recently demonstrated that endothelin-1 (ET-1) activates two types of Ca(2+) permeable nonselective cation channel (designated NSCC-1 and NSCC-2) in Chinese hamster ovarian cells expressing endothelin(B) receptor (CHO-ET(B)R). These channels can be discriminated using the Ca(2+) channel blockers, LOE 908 and SK&F 96365. LOE 908 is a blocker of NSCC-1 and NSCC-2, whereas SK&F 96365 is a blocker of NSCC-2. In this study, we investigated the possible role of phosphoinositide 3 kinase (PI3K) in the ET-1-induced activation of NSCCs in CHO-ET(B)R using wortmannin and LY-294002, inhibitors of PI3K. ET-1-induced Ca(2+) influx was partially inhibited in CHO-ET(B)R pretreated with wortmannin or LY-294002. In contrast, addition of wortmannin or LY-294002 after stimulation with ET-1 did not suppress Ca(2+) influx. The Ca(2+) channels activated by ET-1 in wortmannin- or LY-294002-treated CHO-ET(B)R were sensitive to LOE 908 and resistant to SK&F 96365. In conclusion, NSCC-2 is stimulated by ET-1 via PI3K-dependent cascade, whereas NSCC-1 is stimulated independently of the PI3K pathway. Moreover, PI3K seems to be required for the initiation of the Ca(2+) entry through NSCC-2 but not for its maintenance. PMID- 12529254 TI - Mechanisms of P2X7 receptor-mediated ERK1/2 phosphorylation in human astrocytoma cells. AB - Astrocytes are involved in normal and pathological brain functions, where they become activated and undergo reactive gliosis. Astrocytes have been shown to respond to extracellular nucleotides via the activation of P2 receptors, either G protein-coupled P2Y receptors or P2X receptors that are ligand-gated ion channels. In this study, we have examined the manner in which activation of the P2X(7) nucleotide receptor, an extracellular ATP-gated ion channel expressed in astrocytes, can lead to the phosphorylation of ERK1/2. Results showed that the P2X(7) receptor agonist 2',3'-O-(4-benzoyl)benzoyl-ATP induced ERK1/2 phosphorylation in human astrocytoma cells overexpressing the recombinant rat P2X(7) receptor (rP2X(7)-R), a response that was inhibited by the P2X(7) receptor antagonist, oxidized ATP. Other results suggest that rP2X(7)-R-mediated ERK1/2 phosphorylation was linked to the phosphorylation of the proline-rich/Ca(2+) activated tyrosine kinase Pyk2, c-Src, phosphatidylinositol 3'-kinase, and protein kinase Cdelta activities and was dependent on the presence of extracellular Ca(2+). These results support the hypothesis that the P2X(7) receptor and its signaling pathways play a role in astrocyte-mediated inflammation and neurodegenerative disease. PMID- 12529255 TI - Reduced inotropic heart response in selenium-deficient mice relates with inducible nitric oxide synthase. AB - Atria from mice fed a selenium-deficient (Se(-)) diet have a diminished beta adrenoceptor-inotropic cardiac response to isoproterenol or norepinephrine compared with atria from mice fed the same diet supplemented with 0.2 mg/kg Se as sodium selenite (Se(+)). This diminished response could be reversed by feeding Se(-) mice the Se(+) diet for 1 wk or by pretreatment with nitric oxide synthase (NOS) inhibitors such as N(G)-monomethyl-l-arginine or aminopyridine. Elevated serum concentrations of nitrite/nitrate as well as a threefold increase in the atrial NOS activity were seen in the Se(-) versus Se(+) mice. Western blotting and indirect immunofluorescence indicated an enhanced expression of inducible NOS in hearts from Se(-) mice. Increased expression and activity of NOS and increased nitrite/nitrate levels from Se(-) mice correlated with an impaired response to beta-adrenoceptor inotropic cardiac stimulation. Elevated nitric oxide levels may account for some of the pathophysiological effects of Se deficiency on the heart. PMID- 12529256 TI - Regional upregulation of Kv2.1-encoded current, IK,slow2, in Kv1DN mice is abolished by crossbreeding with Kv2DN mice. AB - Overexpression of a truncated Kv1.1 channel transgene in the heart (Kv1DN) resulted in mice with a prolonged action potential duration due to marked attenuation of a rapidly activating, slowly inactivating potassium current (I(K,slow1)) in ventricular myocytes. Optical mapping and programmed electrical stimulation revealed inducible ventricular tachycardia due to spatial dispersion of repolarization and refractoriness. Here we show that a delayed rectifier with slower inactivation kinetics (I(K,slow2)) was selectively upregulated in Kv1DN cardiocytes. This electrical remodeling was spatially restricted to myocytes derived from the apex of the left ventricle. Biophysical and pharmacological studies of I(K,slow2) indicate that it resembles Kv2-encoded currents. Northern blot analyses and real-time PCR revealed upregulation of Kv2.1 transcript in Kv1DN mice. Crossbreeding of Kv1DN mice with mice expressing a truncated Kv2.1 polypeptide (Kv2DN) eliminated I(K,slow2). In summary, our data indicate that the spatially restrictive upregulation of Kv2.1-encoded currents underlies the increased dispersion of the repolarization observed in Kv1DN mice. PMID- 12529257 TI - OxLDL stimulates cell proliferation through a general induction of cell cycle proteins. AB - Oxidized low-density lipoprotein (oxLDL) may be involved in atherosclerosis by stimulating proliferation of cells in the vessel wall. The purpose of this study was to identify the mechanism by which oxLDL induces proliferation. Quiescent human fibroblasts and rabbit smooth muscle cells were treated with 0, 10, or 50 microg/ml oxLDL for 24-48 h. This resulted in significant increases in total cell counts at both concentrations of oxLDL, at both time points, for both types of cells. Western blot analysis revealed that oxLDL-stimulated cell proliferation was associated with significant increases in the expression of proteins that regulate entry into and progression through the cell cycle [cell division cycle 2, cyclin-dependent kinase (cdk) 2, cdk 4, cyclin B1, cyclin D1, and PCNA]. Surprisingly, the expression of cell cycle inhibitors (p21 and p27) was stimulated by oxLDL as well, but this was to a lesser extent than the effects on cell cycle-activating proteins. OxLDL also induced nuclear localization of all cell cycle proteins examined. The similar effects of oxLDL on the translocation and expression of both cell cycle-activating and -inhibiting proteins may explain the controlled proliferative phenomenon observed in atherosclerosis as opposed to the more rapid proliferative event characteristic of cancer. PMID- 12529258 TI - The novel serine protease PreR-Co promotes endothelium-independent vasorelaxation in rabbit aortic rings. AB - The effect of a novel enzyme (PreR-Co) that activates renal prorenin was studied on rabbit aortas with and without endothelium. It was tested 1) in the basal tone of nonstimulated or ANG II-sensitized rings or rings precontracted with norepinephrine (NE), PGF(2alpha), high KCl concentration, and 2) in rings pretreated with enalaprilat, losartan, PD-123319, N(omega)-nitro-l-arginine methyl ester, HOE-140, indomethacin, or serine protease inhibitors (PMSF, aprotinin, or soybean trypsin inhibitor); kallilkrein and bradykinin were also tested in ANG II-sensitized rings. PreR-Co produced a vasorelaxant effect in the basal tone and in the precontracted rabbit aorta. The effect was endothelium independent, potentiated by endothelium removal or nitric oxide (NO) synthase inhibition, and abolished by boiling the enzyme. In addition, the effect improved when basal tone was increased in ANG II-sensitized aortic rings or in precontracted vessels. No activation of the ANG II, bradykinin, prostaglandin, or NO pathway mediating the PreR-Co response could be obtained, suggesting a direct action of the enzyme. This action seems to be dependent on esterasic activity because serine protease inhibitors like PMSF and aprotinin were able to block the vasorelaxant effect of PreR-Co. PMID- 12529259 TI - Inhibition of vascular ATP-sensitive K+ channels does not affect reactive hyperemia in human forearm. AB - The extent to which ATP-sensitive K(+) channels contribute to reactive hyperemia in humans is unresolved. We examined the role of ATP-sensitive K(+) channels in regulating reactive hyperemia induced by 5 min of forearm ischemia. Thirty-one healthy subjects had forearm blood flow measured with venous occlusion plethysmography. Reactive hyperemia could be reproducibly induced (n = 9). The contribution of vascular ATP-sensitive K(+) channels to reactive hyperemia was determined by measuring forearm blood flow before and during brachial artery infusion of glibenclamide, an ATP-sensitive K(+) channel inhibitor (n = 12). To document ATP-sensitive K(+) channel inhibition with glibenclamide, coinfusion with diazoxide, an ATP-sensitive K(+) channel opener, was undertaken (n = 10). Glibenclamide did not significantly alter resting forearm blood flow or the initial and sustained phases of reactive hyperemia. However, glibenclamide attenuated the hyperemic response induced by diazoxide. These data suggest that ATP-sensitive K(+) channels do not play an important role in controlling forearm reactive hyperemia and that other mechanisms are active in this adaptive response. PMID- 12529260 TI - Intraepithelial lymphocyte-derived interferon-gamma evokes enterocyte apoptosis with parenteral nutrition in mice. AB - Total parenteral nutrition (TPN) results in an increase in intraepithelial lymphocyte (IEL)-derived interferon-gamma (IFN-gamma) expression as well as an increase in epithelial cell (EC) apoptosis. This study examined the role that IEL derived IFN-gamma has in the increase in EC apoptosis. Mice received either TPN or oral feedings for 7 days. Small bowel EC apoptosis significantly rose in mice receiving TPN. The administration of TPN also significantly increased IEL-derived IFN-gamma and Fas ligand (FasL) expression. EC apoptosis in IFN-gamma knockout (IFNKO) mice that received TPN was significantly lower than in wild-type TPN mice. Sensitivity of EC to Fas-mediated apoptosis in IFNKO mice was significantly lower than in wild-type TPN mice. Apoptosis in Fas-deficient and FasL-deficient mice that received TPN was significantly lower than in wild-type mice that received TPN. The TPN-induced increase in IFN-gamma expression appears to result in an increase in Fas-L expression and EC sensitivity to Fas, with a resultant increase in EC apoptosis. This may well be one of the mediators of increased EC apoptosis observed with TPN administration. PMID- 12529261 TI - Novel MUC1 splice variants contribute to mucin overexpression in CFTR-deficient mice. AB - A cystic fibrosis (CF) mouse expressing the human mucin MUC1 transgene (CFM) reverted the CF/Muc1(-/-) phenotype (little mucus accumulated in the intestine) to that of CF mice expressing mouse Muc1, which exhibited increased mucus accumulation. Western blots and immunohistochemical analysis showed that the MUC1 protein was markedly increased in CFM mice in which it was both membrane bound and secreted into the intestinal lumen. Studies to determine the reason for increased levels of the extracellular domain of MUC1 mucin identified mRNA and protein of two novel splice variants and the previously described secreted MUC1 lacking the cytoplasmic tail (MUC1/SEC). Novel MUC1 splice variants, CT80 and CT58, were both transmembrane proteins with cytoplasmic tails different from the normal MUC1. The MUC1-CT80 and MUC1/SEC forms are found expressed mainly in the CFM mice intestines. Thus MUC1 expression is increased, and it appears that alternate cytoplasmic tails may change its role in signaling. MUC1 could be an important contributor to the CF intestinal phenotype. PMID- 12529262 TI - Hepatic inflammatory mediators contribute to intestinal damage in necrotizing enterocolitis. AB - Necrotizing enterocolitis (NEC) is a common and devastating gastrointestinal disease of premature infants. Along with pathological effects in the ileum, severe NEC is often accompanied by multisystem organ failure, including liver failure. The aim of this study was to determine the changes in hepatic cytokines and inflammatory mediators in experimental NEC. The well-established neonatal rat model of NEC was used in this study, and changes in liver morphology, numbers of Kupffer cells (KC), gene expression, and histological localization of IL-18, TNF alpha, and inducible nitric oxide synthase were evaluated. Intestinal luminal TNF alpha levels were also measured. Production of hepatic IL-18 and TNF-alpha and numbers of KC were increased in rats with NEC and correlated with the progression of intestinal damage during NEC development. Furthermore, increased levels of TNF alpha in the intestinal lumen of rats with NEC was significantly decreased when KC were inhibited with gadolinium chloride. These results suggest an important role of the liver and the gut-liver axis in NEC pathogenesis. PMID- 12529263 TI - The water load test: observations from healthy controls and patients with functional dyspepsia. AB - Gastric sensation and accommodation are studied by barostat, but this is invasive. The drink test is noninvasive and may provide similar information. We evaluated relationships between drink test, gastric function, symptoms, and psychiatric distress. Controls (73) and functional dyspeptics (FD) (92) were studied using a 5-min water load test (WL5), gastric emptying, and electrogastrography (EGG). Symptoms, quality of life, and psychiatric distress were measured using standardized measures. Controls underwent test-retest of WL5 and comparison of WL5 with 100 ml/min water-based drink test (WL100) or nutrient drink. Controls, FD, and gastroparetics estimated drinking capacity before WL5 using a visual analog scale. WL5 correlated with WL100 (r = 0.7929) but not nutrient drink test (r = 0.1995). WL5 was significantly less in FD than controls, and abnormal WL5 was seen in 46%. In FD, volume to fullness inversely correlated with symptom severity (r =-0.29; P = 0.0154) and WL5 produced more symptoms, particularly nausea. Gastric function was not different between FD with normal or abnormal WL5. Symptoms and psychiatric distress were similar between normal and abnormal WL5 groups, but the abnormal group had significantly poorer quality of life. Controls and gastroparetics had good correlation of estimated and ingested volumes, but FD did not. Versus FD with normal WL5 capacity, FD with impaired drinking capacity have normal gastric function and similar symptoms but poorer quality of life. FD are less able to predict drinking capacity. These data suggest that WL5 identifies FD with intact gastric function but abnormal visceral perception. PMID- 12529264 TI - Relative contribution of various airway protective mechanisms to prevention of aspiration during swallowing. AB - Deglutitive airway protective mechanisms include glottal closure, epiglottal descent, and anterosuperior displacement of the larynx. Aspiration of swallowed material may occur during the pre-, intra-, or postpharyngeal phase of swallowing. Our objectives were to determine the relative contribution of the airway protective mechanisms during each phase of swallow in 14 decerebrated cats before and after suprahyoid myotomy, epiglottectomy, and unilateral cordectomy. After myotomy, superior excursions of the hyoid, thyroid, and cricoid cartilages and anteroposterior diameter of maximum upper esophageal spincter (UES) opening were significantly diminished, but the incidence of pharyngeal residue significantly increased (P < 0.05). No aspiration was observed in the predeglutitive period. After myotomy, the incidence of aspiration significantly increased in both intra- and postdeglutitive periods. Epiglottectomy did not alter aspiration incidence, but unilateral cordectomy resulted in a 100% incidence of intra- and postdeglutitive aspiration. In conclusion, glottal closure constitutes the primary mechanism for prevention of intra- and postdeglutitive aspiration, but laryngeal elevation may assist this function. Bolus pulsion without laryngeal distraction can open the UES, but at risk of aspiration due to decreased pharyngeal clearance. The epiglottis provides no apparent airway protection during any phase of swallowing. PMID- 12529265 TI - Bile acid regulation of hepatic physiology: I. Hepatocyte transport of bile acids. AB - Bile acids are cholesterol derivatives that serve as detergents in bile and the small intestine. Approximately 95% of bile acids secreted by hepatocytes into bile are absorbed from the distal ileum into the portal venous system. Extraction from the portal circulation by the hepatocyte followed by reexcretion into the bile canaliculus completes the enterohepatic circulation of these compounds. Over the past few years, candidate bile acid transport proteins of the sinusoidal and canalicular plasma membranes of the hepatocyte have been identified. The physiology of hepatocyte bile acid transport and its relationship to these transport proteins is the subject of this Themes article. PMID- 12529266 TI - Musings on the wanderer: what's new in our understanding of vago-vagal reflexes? III. Activity-dependent plasticity in vago-vagal reflexes controlling the stomach. AB - Vago-vagal reflex circuits modulate digestive functions from the oral cavity to the transverse colon. Previous articles in this series have described events at the level of the sensory receptors encoding the peripheral stimuli, the transmission of information in the afferent vagus, and the conversion of this data within the dorsal vagal complex (DVC) to impulses in the preganglionic efferents. The control by vagal efferents of the postganglionic neurons impinging on the glands and smooth muscles of the target organs has also been illustrated. Here we focus on some of the mechanisms by which these apparently static reflex circuits can be made quite plastic as a consequence of the action of modulatory inputs from other central nervous system sources. A large body of evidence has shown that the neuronal elements that constitute these brain stem circuits have nonuniform properties and function differently according to status of their target organs and the level of activity in critical modulatory inputs. We propose that DVC circuits undergo a certain amount of short-term plasticity that allows the brain stem neuronal elements to act in harmony with neural systems that control behavioral and physiological homeostasis. PMID- 12529267 TI - Selective phosphorylation of the IP3R-I in vivo by cGMP-dependent protein kinase in smooth muscle. AB - This study examined the expression of inositol 1,4,5-trisphosphate (IP(3)) receptor (IP(3)R) types and PKG isoforms in isolated gastric smooth muscle cells and determined the ability of PKG and PKA to phosphorylate IP(3)Rs and inhibit IP(3)-dependent Ca(2+) release, which mediates the initial phase of agonist induced contraction. PKG-Ialpha and PKG-Ibeta were expressed in gastric smooth muscle cells, together with IP(3)-R-associated cG-kinase substrate, a protein that couples PKG-Ibeta to IP(3)R-I. IP(3)R-I and IP(3)R-III were also expressed, but only IP(3)R-I was phosphorylated by PKA and PKG in vitro and exclusively by PKG in vivo. Sequential phosphorylation by PKA and by PKG-Ialpha in vitro showed that PKA phosphorylated the same site as PKG (presumably S(1755)) and an additional PKA-specific site (S(1589)). In intact muscle cells, agents that activated PKG or both PKG and PKA induced IP(3)R-I phosphorylation that was reversed by the PKG inhibitor (8R,9S,11s)-(-)-9-methoxy-carbamyl-8-methyl 2,3,9,10-tetrahydro-8,11-epoxy-1H,8H,1H,-2,7b,11a-trizadizo benzo9(a,g)cycloocta(c,d,e)-trinden-1-one. Agents that activated PKA induced IP(3)R-I phosphorylation in permeabilized but not intact muscle cells, implying that PKA does not gain access to IP(3)R-I in intact muscle cells. The pattern of IP(3)R-I phosphorylation in vivo and in vitro was more consistent with phosphorylation by PKG-Ialpha. Phosphorylation of IP(3)R-I in microsomes by PKG, PKA, or a combination of PKG and PKA inhibited IP(3)-induced Ca(2+) release to the same extent, implying that inhibition was mediated by phosphorylation of the PKG-specific site. We conclude that IP(3)R-I is selectively phosphorylated by PKG I in intact smooth muscle resulting in inhibition of IP(3)-dependent Ca(2+) release. PMID- 12529268 TI - Increases in biliary cholesterol-to-bile acid ratio in pregnant hamsters fed low and high levels of cholesterol. AB - Gallstones develop when the secretion of cholesterol is elevated compared with the secretion of bile acids into bile. One of the risk factors for the formation of gallstones is pregnancy. Because the pregnancy-induced increase in hepatic cholesterol synthesis rates could play a critical role in the development of cholesterol stones, the aim of the present study was to determine whether stone formation, as assessed by the ratio of cholesterol to bile acids in bile, could be ablated by blocking the pregnancy-induced increase in hepatic sterol synthesis rates. Golden Syrian hamsters were fed either ground chow or chow supplemented with 0.5% cholesterol for 3 wk and studied in the nonpregnant state or in late gestation. In chow-fed animals, a 1.6-fold increase in the ratio of cholesterol to bile acids occurred simultaneously with a sevenfold increase in hepatic sterol synthesis rate and a ninefold increase in the amount of newly synthesized cholesterol secreted into the bile in late gestation. In the cholesterol-fed dams, an increase in the ratio of cholesterol to bile acids occurred even with the lack of induction of hepatic sterol synthesis rates during pregnancy. Thus it appears that the marked induction of hepatic sterol synthesis rates during gestation is not essential for the pregnancy-induced cholesterol saturation of bile when cholesterol is fed to animals. PMID- 12529269 TI - Prepro-endothelin-1 mRNA and its mature peptide in human appendix. AB - Because the precise immunopathological events occurring in appendicitis are not completely understood, possible local production of endothelin-1 (ET-1) in human appendix was investigated. We used immunohistochemistry and in situ hybridization to detect the presence, distribution, and phenotype of ET-1-positive cells and prepro-ET-1 (pp-ET-1) mRNA-expressing cells. ET-1-positive stromal cells and pp ET-1 mRNA-expressing cells were detected with different distributions and relative frequencies in normal control appendix, histologically normal appendix, and inflamed appendix. Six of 20 histologically normal appendixes from patients with a clinical diagnosis of acute appendicitis had many ET-1-positive stromal cells and high pp-ET-1 mRNA expression, similar to inflamed appendix. Forty percent of the pp-ET-1 mRNA-expressing cells were neutrophils, and the other positive cells were mast cells and macrophages. We suggest that local production of ET-1 by neutrophils and other inflammatory cells could be a molecular sign of focal inflammation in histologically normal appendixes and that ET-1 could be implicated, with other cytokines, in the pathogenesis of appendicitis by inducing appendiceal ischemia through vasoconstriction. PMID- 12529270 TI - TGF-beta signal transduction and mesangial cell fibrogenesis. AB - Transforming growth factor-beta (TGF-beta) is closely associated with progressive renal fibrosis. Significant progress has been accomplished in determining the cellular signaling pathways that are activated by TGF-beta. This knowledge is being applied to glomerular mesangial cell models of extracellular matrix (ECM) accumulation. A central component of TGF-beta-stimulated mesangial cell fibrogenesis is the TGF-beta family-specific Smad signal transduction pathway. However, while Smads play an important role in collagen accumulation, recent findings indicate that cross talk among a variety of pathways is necessary for maximal stimulation of collagen expression. Further investigation of these multiple interactions will provide insight into possible ways to interrupt cellular mechanisms of glomerular fibrogenesis. PMID- 12529271 TI - Physiology of the renal medullary microcirculation. AB - Perfusion of the renal medulla plays an important role in salt and water balance. Pericytes are smooth muscle-like cells that impart contractile function to descending vasa recta (DVR), the arteriolar segments that supply the medulla with blood flow. DVR contraction by ANG II is mediated by depolarization resulting from an increase in plasma membrane Cl(-) conductance that secondarily gates voltage-activated Ca(2+) entry. In this respect, DVR may differ from other parts of the efferent microcirculation of the kidney. Elevation of extracellular K(+) constricts DVR to a lesser degree than ANG II or endothelin-1, implying that other events, in addition to membrane depolarization, are needed to maximize vasoconstriction. DVR endothelial cytoplasmic Ca(2+) is increased by bradykinin, a response that is inhibited by ANG II. ANG II inhibition of endothelial Ca(2+) signaling might serve to regulate the site of origin of vasodilatory paracrine agents generated in the vicinity of outer medullary vascular bundles. In the hydropenic kidney, DVR plasma equilibrates with the interstitium both by diffusion and through water efflux across aquaporin-1. That process is predicted to optimize urinary concentration by lowering blood flow to the inner medulla. To optimize urea trapping, DVR endothelia express the UT-B facilitated urea transporter. These and other features show that vasa recta have physiological mechanisms specific to their role in the renal medulla. PMID- 12529272 TI - Platelet-activating factor and solute transport processes in the kidney. AB - We examined the hemodynamic and tubular transport mechanisms by which platelet activating factor (PAF) regulates salt and water excretion. In anesthetized, renally denervated male Wistar rats, with raised systemic blood pressure and renal arterial blood pressure maintained at normal levels, intrarenal PAF infusion at 2.5 ng. min(-1) x kg(-1) resulted in a small fall in systemic blood pressure (no change in renal arterial blood pressure) and an increase in renal blood flow and urinary water, sodium, and potassium excretion rates. The PAF induced changes in cardiovascular and renal hemodynamic function were abolished and renal excretory function greatly attenuated by treating rats with a nitric oxide synthase inhibitor. To determine whether a tubular site of action was involved in the natriuretic effect of PAF, cortical proximal tubules were enzymatically dissociated from male Wistar rat kidneys, and oxygen consumption rates (Qo(2)) were used as an integrated index of transcellular sodium transport. PAF at 1 nM maximally inhibited Qo(2) in both untreated and nystatin-stimulated (sodium entry into renal cell is not rate limiting) proximal tubules by approximately 20%. Blockade of PAF receptors or Na(+)-K(+)-ATPase pump activity with BN-52021 or ouabain, respectively, abolished the effect of PAF on nystatin stimulated proximal tubule Qo(2). Inhibition of nitric oxide synthase or guanylate cyclase systems did not alter PAF-mediated inhibition of nystatin stimulated proximal tubule Qo(2), whereas phospholipase A(2) or cytochrome-P-450 monooxygenase inhibition resulted in a 40-60% reduction. These findings suggest that stimulation of PAF receptors on the proximal tubule decreases transcellular sodium transport by activating phospholipase A(2) and the cytochrome-P-450 monooxygenase pathways that lead to the inhibition of an ouabain-sensitive component of the basolateral Na(+)-K(+)-ATPase pump. Thus PAF can activate both an arachidonate pathway-mediated suppression of proximal tubule sodium transport and a nitric oxide pathway-mediated dilatory action on renal hemodynamics that likely contributes to the natriuresis and diuresis observed in vivo. PMID- 12529273 TI - Delayed administration of hepatocyte growth factor reduces renal fibrosis in obstructive nephropathy. AB - Hepatocyte growth factor (HGF) is a renotropic protein that elicits antifibrogenic activity by preventing the activation of matrix-producing myofibroblast cells in animal models of chronic renal diseases. However, whether a delayed administration of HGF can still attenuate renal fibrosis remains uncertain. In this study, we examined the therapeutic potential of exogenous HGF on an established renal interstitial fibrosis induced by unilateral ureteral obstruction (UUO). Three days after UUO, the obstructed kidneys displayed interstitial fibrotic lesions with characteristic features of an established renal fibrosis, as manifested by myofibroblast activation, fibronectin overexpression, interstitial matrix deposition, and transforming growth factor beta1 upregulation. Beginning at this time point, administration of recombinant HGF into mice by intravenous injections for 11 days markedly suppressed the progression of renal interstitial fibrosis. HGF significantly suppressed renal alpha-smooth muscle actin expression, total kidney collagen contents, interstitial matrix components, such as fibronectin, and renal expression of transforming growth factor-beta1 and its type I receptor. Compared with the starting point (3 days after UUO), HGF treatment largely blunted the progression of myofibroblast accumulation and collagen deposition but did not reverse it. Delayed administration of HGF also suppressed the myofibroblastic transdifferentiation from tubular epithelial cells in vitro, as demonstrated by a decline in alpha-smooth muscle actin and fibronectin expression. These results suggest that exogenous HGF exhibits potent therapeutic effects on retarding the progression of an established renal fibrosis. PMID- 12529274 TI - Angiotensin II and renal medullary blood flow in Lyon rats. AB - The present study evaluated the acute effects of ANG II (5-480 ng/kg iv) and phenylephrine (PE; 0.2-146 microg/kg iv) on total renal (RBF) and medullary blood flow (MBF) in anesthetized Lyon hypertensive (LH) and low-blood-pressure (LL) rats. ANG II and PE induced dose-dependent decreases in both RBF and MBF, which were greater in LH than in LL rats. Interestingly, after ANG II, but not after PE, the initial medullary vasoconstriction was followed by a long-lasting and dose-dependent vasodilation that was significantly blunted in LH compared with LL rats. The mechanisms of the MBF effects of ANG II were studied in LL rats only. Blockade of AT(1) receptors with losartan (10 mg/kg) abolished all the effects of ANG II, whereas AT(2) receptor blockade with PD-123319 (50 microg x kg(-1) x min( 1) iv) did not change these effects. Indomethacin (5 mg/kg) decreased by approximately 90% the medullary vasodilation induced by the lowest doses of ANG II (from 15 ng/kg). In contrast, N(G)-nitro-l-arginine methyl ester (10 mg/kg and 0.1 mg. kg(-1). min(-1) iv) and the bradykinin B(2)-receptor antagonist HOE-140 (20 microg/kg and 10 microg x kg(-1) x min(-1) iv) markedly lowered the medullary vasodilation at the highest doses of ANG II only. In conclusion, this study shows that LH rats exhibit an altered MBF response to ANG II compared with LL rats and indicates that the AT(1) receptor-mediated medullary vasodilator response to low doses of ANG II is mainly due to the release of PGs, whereas the dilator response to high doses of ANG II has additional nitric oxide- and kinin-dependent components. PMID- 12529275 TI - Role of luminal anion and pH in distal tubule potassium secretion. AB - Potassium secretory flux (J(K)) by the distal nephron is regulated by systemic and luminal factors. In the present investigation, J(K) was measured with a double-barreled K(+) electrode during paired microperfusion of superficial segments of the rat distal nephron. We used control solutions (100 mM NaCl, pH 7.0) and experimental solutions in which Cl(-) had been replaced with a less permeant anion and/or pH had been increased to 8.0. J(K) increased when Cl(-) was replaced by either acetate ( approximately 37%), sulfate ( approximately 32%), or bicarbonate ( approximately 62%), and also when the pH of the control perfusate was increased ( approximately 26%). The majority (80%) of acetate-stimulated J(K) was Ba(2+) sensitive, but furosemide (1 mM) further reduced secretion ( approximately 10% of total), suggesting that K(+)-Cl(-) cotransport was operative. Progressive reduction in luminal Cl(-) concentration from 100 to 20 to 2 mM caused increments in J(K) that were abolished by inhibitors of K(+)-Cl(-) cortransport, i.e., furosemide and [(dihydroindenyl)oxy]alkanoic acid. Increasing the pH of the luminal perfusion fluid also increased J(K) even in the presence of Ba(2+), suggesting that this effect cannot be accounted for only by K(+) channel modulation of K(+) secretion in the distal nephron of the rat. Collectively, these data suggest a role for K(+)-Cl(-) cotransport in distal nephron K(+) secretion. PMID- 12529276 TI - Reversibility of chronic cyclosporine nephropathy in rats after withdrawal of cyclosporine. AB - Renal interstitial inflammation is an important factor in the pathogenesis of chronic cyclosporin A (CsA) nephropathy. We studied the expression of the chemoattractant osteopontin (OPN) and the relationship between OPN expression and tubulointerstitial injury in a rat model of chronic CsA nephropathy. Chronic CsA nephropathy was induced in Sprague-Dawley rats by administering CsA (15 mg/kg sc) for 5 wk and then withdrawing it for 5 or 10 wk. Renal function, histopathology (arteriolopathy, ED-1-positive cells, and tubulointerstitial fibrosis), renin angiotensin system (RAS) activity, and OPN expression were observed during the follow-up period. Renal function deteriorated in CsA-treated rats, with the development of typical histopathology and activation of RAS. After CsA withdrawal, these parameters were significantly reversed (all P < 0.05). The upregulation of OPN mRNA and protein expression seen in CsA-treated rat kidneys was decreased 5 wk after CsA withdrawal and was further decreased after 10 wk. Of note, OPN mRNA expression correlated with the number of infiltrating macrophage (r = 0.651, P < 0.01) and tubulointerstitial fibrosis (r = 0.729, P < 0.01). These findings suggest that OPN expression and macrophage infiltration decrease after long-term CsA withdrawal in rats with established chronic CsA nephropathy, and this is closely associated with recovery from renal injury. PMID- 12529277 TI - Differential, inducible gene targeting in renal epithelia, vascular endothelium, and viscera of Mx1Cre mice. AB - The Cre/loxP transgenic system may be used to achieve temporally and/or spatially regulated gene deletion. The Mx1Cre mouse expresses Cre recombinase under control of the IFN-inducible Mx1 promoter. Mx1Cre mice were crossed with a reporter strain (ROSA26tm1Sor) in which beta-galactosidase activity is expressed only after Cre-mediated recombination to determine the cellular pattern of Cre mediated genetic recombination in the kidney and other tissues. Widespread recombination was observed in vascular endothelium as well as in the liver and spleen. Recombination was restricted to subsets of stromal cells in uterus, duodenum, colon, aorta, and kidney. In the cortex, chi-galactosidase activity was detected in a subset of tubules and all glomerular cells, including endothelium, mesangium, and podocytes. No chi-galactosidase activity was detected in proximal tubules. Costaining of kidneys with segment-specific markers demonstrated induction of chi-galactosidase activity in collecting duct, with sporadic labeling of the thick ascending limb but no significant labeling of distal convoluted tubules. We conclude that Mx1-driven gene recombination is spatially as well as temporally restricted. The Mx1Cre transgene should prove a useful reagent to achieve temporally regulated recombination in endothelial, glomerular, and distal renal epithelia in mice. PMID- 12529278 TI - Possible mechanisms underlying pregnancy-induced changes in uterine artery endothelial function. AB - The last 10 years has seen a dramatic increase in our understanding of the mechanisms underlying the pregnancy-specific adaptation in cardiovascular function in general and the dramatic changes that occur in uterine artery endothelium in particular to support the growing fetus. The importance of these changes is clear from a number of studies linking restriction of uterine blood flow (UBF) and/or endothelial dysfunction and clinical conditions such as intrauterine growth retardation (IUGR) and/or preeclampsia in both humans and animal models; these topics are covered only briefly here. The recent developments that prompts this review are twofold. The first is advances in an understanding of the cell signaling processes that regulate endothelial nitric oxide synthase (eNOS) in particular (Govers R and Rabelink TJ. Am J Physiol Renal Physiol 280: F193-F206, 2001). The second is the emerging picture that uterine artery (UA) endothelial cell production of nitric oxide (NO) as well as prostacyclin (PGI2) may be as much a consequence of cellular reprogramming at the level of cell signaling as due to tonic stimuli inducing changes in the level of expression of eNOS or the enzymes of the PGI2 biosynthetic pathway (cPLA2, COX-1, PGIS). In reviewing just how we came to this conclusion and outlining the implications of such a finding, we draw mostly on data from ovine or human studies, with reference to other species only where directly relevant. PMID- 12529279 TI - Heart failure and the brain: new perspectives. AB - Despite recent therapeutic advances, the prognosis for patients with heart failure remains dismal. Unchecked neurohumoral excitation is a critical element in the progressive clinical deterioration associated with the heart failure syndrome, and its peripheral manifestations have become the principal targets for intervention. The link between peripheral systems activated in heart failure and the central nervous system as a source of neurohumoral drive has therefore come under close scrutiny. In this context, the forebrain and particularly the paraventricular nucleus of the hypothalamus have emerged as sites that sense humoral signals generated peripherally in response to the stresses of heart failure and contribute to the altered volume regulation and augmented sympathetic drive that characterize the heart failure syndrome. This brief review summarizes recent studies from our laboratory supporting the concept that the forebrain plays a critical role in the pathogenesis of ischemia-induced heart failure and suggesting that the forebrain contribution must be considered in designing therapeutic strategies. Forebrain signaling by neuroactive products of the renin angiotensin system and the immune system are emphasized. PMID- 12529280 TI - Thermoregulation. PMID- 12529281 TI - Individual differences in subjective and objective alertness during sleep deprivation are stable and unrelated. AB - This study examines the individual reproducibility of alterations of subjective, objective, and EEG measures of alertness during 27 h of continuous wakefulness and analyzes their interrelationships. Eight subjects were studied twice under similar constant-routine conditions. Scales and performance tasks were administered at hourly intervals to define temporal changes in subjective and objective alertness. The wake EEG was recorded every 2 h, 2 min with eyes open and 2 min with eyes closed. Plasma glucose and melatonin levels were measured to estimate brain glucose utilization and individual circadian phase, respectively. Decrements of subjective alertness and performance deficits were found to be highly reproducible for a given individual. Remarkably, there was no relationship between the impairments of subjective and objective alertness. With increased duration of wakefulness, EEG activity with eyes closed increased in the delta range and decreased in the alpha range, but the magnitudes of these changes were also unrelated. These findings indicate that sleep deprivation has highly reproducible, but independent, effects on brain mechanisms controlling subjective and objective alertness. PMID- 12529282 TI - Central mineralocorticoid receptor blockade decreases plasma TNF-alpha after coronary artery ligation in rats. AB - The Randomized Aldactone Evaluation Study (RALES) demonstrated a substantial clinical benefit to blocking the effects of aldosterone (Aldo) in patients with heart failure. We recently demonstrated that the enhanced renal conservation of sodium and water in rats with heart failure can be reduced by blocking the central nervous system effects of Aldo with the mineralocorticoid receptor (MR) antagonist spironolactone (SL). Preliminary data from our laboratory suggested that central MR might contribute to another peripheral mechanism in heart failure, the release of proinflammatory cytokines. In the present study, SL (100 ng/h for 21 days) or ethanol vehicle (Veh) was administered via the 3(rd) cerebral ventricle to one group of rats after coronary ligation (CL) or sham CL (Sham) to induce congestive heart failure (CHF). In Veh-treated CHF rats, tumor necrosis factor-alpha (TNF-alpha) levels increased during day 1 and continued to increase throughout the 3-wk observation period. In CHF rats treated with SL, started 24 h after CL, TNF-alpha levels rose initially but retuned to control levels by day 5 after CL and remained low throughout the study. These findings suggest that activation of MR in the central nervous system plays a critical role in regulating TNF-alpha release in heart failure rats. Thus some of the beneficial effect of blocking MR in heart failure could be due at least in part to a reduction in TNF-alpha production. PMID- 12529283 TI - Endogenous cannabinoids: a new target in the treatment of obesity. PMID- 12529284 TI - Artificial selection for high activity favors mighty mini-muscles in house mice. AB - After 14 generations of selection for voluntary wheel running, mice from the four replicate selected lines ran, on average, twice as many revolutions per day as those from the four unselected control lines. To examine whether the selected lines followed distinct strategies in the correlated responses of the size and metabolic capacities of the hindlimb muscles, we examined mice from selected lines, housed for 8 wk in cages with access to running wheels that were either free to rotate ("wheel access" group) or locked ("sedentary"). Thirteen of twenty individuals in one selected line (line 6) and two of twenty in another (line 3) showed a marked reduction ( approximately 50%) in total hindlimb muscle mass, consistent with the previously described expression of a small-muscle phenotype. Individuals with these "mini-muscles" were not significantly smaller in total body mass compared with line-mates with normal-sized muscles. Access to free wheels did not affect the relative mass of the mini-muscles, but did result in typical mammalian training effects for mitochondrial enzyme activities. Individuals with mini-muscles showed a higher mass-specific muscle aerobic capacity as revealed by the maximal in vitro rates of citrate synthase and cytochrome c oxidase. Moreover, these mice showed the highest activities of hexokinase and carnitine palmitoyl transferase. Females with mini-muscles showed the highest levels of phosphofructokinase, and males with mini-muscles the highest levels of pyruvate dehydrogenase. As shown by total muscle enzyme contents, the increase in mass-specific aerobic capacity almost completely compensated for the reduction caused by the "loss" of muscle mass. Moreover, the mini-muscle mice exhibited the lowest contents of lactate dehydrogenase and glycogen phosphorylase. Interestingly, metabolic capacities of mini-muscled mice resemble those of muscles after endurance training. Overall, our results demonstrate that during selection for voluntary wheel running, distinct adaptive paths that differentially exploit the genetic variation in morphological and physiological traits have been followed. PMID- 12529285 TI - Influence of respiratory network drive on phrenic motor output evoked by activation of cat pre-Botzinger complex. AB - We have previously demonstrated that microinjection of dl-homocysteic acid (DLH), a glutamate analog, into the pre-Botzinger complex (pre-BotC) can produce either phasic or tonic excitation of phrenic nerve discharge during hyperoxic normocapnia. Breathing, however, is influenced by input from both central and peripheral chemoreceptor activation. This influence of increased respiratory network drive on pre-BotC-induced modulation of phrenic motor output is unclear. Therefore, these experiments were designed to examine the effects of chemical stimulation of neurons (DLH; 10 mM; 10-20 nl) in the pre-BotC during hyperoxic modulation of CO2 (i.e., hypercapnia and hypocapnia) and during normocapnic hypoxia in chloralose-anesthetized, vagotomized, mechanically ventilated cats. For these experiments, sites were selected in which unilateral microinjection of DLH into the pre-BotC during baseline conditions of hyperoxic normocapnia [arterial PCO2 (PaCO2) = 37-43 mmHg; n = 22] produced a tonic (nonphasic) excitation of phrenic nerve discharge. During hypercapnia (PaCO2 = 59.7 +/- 2.8 mmHg; n = 17), similar microinjection produced excitation in which phasic respiratory bursts were superimposed on varying levels of tonic discharge. These DLH-induced phasic respiratory bursts had an increased frequency compared with the preinjection baseline frequency (P < 0.01). In contrast, during hypocapnia (PaCO2 = 29.4 +/- 1.5 mmHg; n = 11), microinjection of DLH produced nonphasic tonic excitation of phrenic nerve discharge that was less robust than the initial (normocapnic) response (i.e., decreased amplitude). During normocapnic hypoxia (PaCO2 = 38.5 +/- 3.7; arterial Po2 = 38.4 +/- 4.4; n = 8) microinjection of DLH produced phrenic excitation similar to that seen during hypercapnia (i.e., increased frequency of phasic respiratory bursts superimposed on tonic discharge). These findings demonstrate that phrenic motor activity evoked by chemical stimulation of the pre-BotC is influenced by and integrates with modulation of respiratory network drive mediated by input from central and peripheral chemoreceptors. PMID- 12529286 TI - Deciphering the physiological roles of COX-2. PMID- 12529287 TI - Influence of oxygen partial pressures on protein synthesis in feeding crabs. AB - Many water-breathing animals have a strategy that consists of maintaining low blood PO2 values in a large range of water oxygenation level (4-40 kPa). This study examines the postprandial changes in O2 consumption, arterial blood PO2, and tissue protein synthesis in the shore crab Carcinus maenas in normoxic, O2 depleted, and O2-enriched waters to study the effects of this strategy on the O2 consumption and peptide bond formation after feeding. In normoxic water (21 kPa), the arterial PO2 was 1.1 kPa before feeding and 1.2 kPa 24 h later. In water with a PO2 of 3 kPa (arterial PO2 0.6 kPa), postprandial stimulation of protein synthesis and O2 consumption were blocked. The blockade was partial at a water PO2 of 4 kPa (arterial PO2 0.8 kPa). An increase in environmental PO2 (60 kPa, arterial PO2 10 kPa) resulted in an increase in protein synthesis compared with normoxic rates. It is concluded that the arterial PO2 spontaneously set in normoxic Carcinus limits the rates of protein synthesis. The rationale for such a strategy is discussed. PMID- 12529289 TI - The role of Id2 and apoptosis during skeletal muscle remodeling. PMID- 12529288 TI - Responses to human CGRP, ADM, and PAMP in human thymic arteries. AB - Responses to human CGRP, adrenomedullin (ADM), and proadrenomedullin NH2-terminal 20 peptide (PAMP) were studied in small human thymic arteries. CGRP, ADM, and PAMP produced concentration-dependent vasodilator responses in arteries preconstricted with the thromboxane mimic U-46619. Responses to ADM and PAMP were attenuated, whereas responses to CGRP were not altered by endothelial denudation. Inhibitors of nitric oxide synthase and guanylyl cyclase attenuated responses to ADM and PAMP but not to CGRP. The CGRP1 receptor antagonist CGRP(8-37) attenuated responses to CGRP and ADM but not to PAMP. Responses to CGRP were reduced by SQ 22536 and Rp-cAMPS, inhibitors of adenylyl cyclase and PKA. These data suggest that responses to CGRP and ADM are mediated by CGRP(8-37)-sensitive receptors and that the endothelial ADM receptor induces vasodilation by a nitric oxide-guanylyl cyclase mechanism, whereas a smooth muscle CGRP receptor signals by a cAMP dependent mechanism. A different endothelial receptor recognizes PAMP and signals by a nitric oxide-dependent mechanism. PMID- 12529290 TI - Pituitary adenylate cyclase-activating polypeptide and adrenomedullary function. PMID- 12529291 TI - Expression of smooth muscle MyHC B in blood vessels of hypertrophied heart in experimentally hypertensive rats. AB - We demonstrated recently a significantly lower fraction of cardiac precapillary arterioles that expressed smooth muscle myosin heavy chain (MyHC) B (SMB) in spontaneously hypertensive rats. To clarify whether this reduction of SMB expression is of genetic origin, we investigated SMB expression in cardiac precapillary arterioles of normotensive and experimentally hypertensive rats (one clip, one kidney or ANG II minipump). We observed similar SMB expression patterns in precapillary arterioles of experimentally hypertensive rats compared with normotensive controls. These observations suggest that the downregulation of SMB in spontaneously hypertensive rats is of genetic origin rather than an adaptive response to chronically enhanced blood pressure and cardiac hypertrophy. PMID- 12529292 TI - Dynamics of GPIIb/IIIa-mediated platelet-platelet interactions in platelet adhesion/thrombus formation on collagen in vitro as revealed by videomicroscopy. AB - The conventional description of platelet interactions with collagen-coated surfaces in vitro, based on serial static measurements, is that platelets first adhere and spread to form a monolayer and then recruit additional layers of platelets. To obtain dynamic information, we studied gravity-driven platelet deposition in vitro on purified type 1 collagen by video phase-contrast microscopy at 22 degrees C. With untreated human and wild-type mouse platelets, soon after the initial adhesion of a small number of "vanguard" platelets, "follower" platelets attached to the spread-out vanguard platelets. Follower platelets then adhered to and spread onto nearby collagen or over the vanguard platelets. Thus, thrombi formed as a concerted process rather than as sequential processes. Treatment of human platelets with monoclonal antibody (mAb) 7E3 (anti GPIIb/IIIa (alphaIIbbeta3) + alphaVbeta3) or tirofiban (anti-GPIIb/IIIa) did not prevent platelet adhesion but nearly eliminated the deposition of follower platelets onto vanguard platelets and platelet thrombi. Similar results were obtained with Glanzmann thrombasthenia platelets. Wild-type mouse platelets in the presence of mAb 1B5 (anti-GPIIb/IIIa) and platelets from beta3-null mice behaved like human platelets in the presence of 7E3 or tirofiban. Deposition patterns of untreated human and wild-type mouse platelets were consistent with random distributions under a Poisson model, but those obtained with 7E3- and tirofiban-treated human platelets, 1B5-treated mouse platelets, or beta3-null platelets demonstrated a more uniform deposition than predicted. Thus, in this model system, absence or blockade of GPIIb/IIIa receptors interferes with thrombus formation and alters the pattern of platelet deposition. PMID- 12529293 TI - Follicular lymphoma grade 3B includes 3 cytogenetically defined subgroups with primary t(14;18), 3q27, or other translocations: t(14;18) and 3q27 are mutually exclusive. AB - Chromosomal translocations involving t(14;18)(q32;q21) and the chromosome 3q27 region are common in B-cell non-Hodgkin lymphoma of germinal center cell origin. Grade 3B follicular lymphoma (FL), consisting almost exclusively of centroblasts, is a distinct subgroup of follicular lymphomas that has more in common clinically with the aggressive diffuse large B-cell lymphomas than with their indolent FL grade 1 and 2 counterparts. We studied the cytogenetic and molecular genetic aberrations by classic cytogenetics, polymerase chain reaction, Southern blot hybridization, and fluorescence in situ hybridization, with special emphasis on t(14;18), affecting bcl-2, and 3q27 rearrangement, affecting bcl-6, in 32 cases of FL grade 3B. Three distinctive subgroups were identified based upon the existence of breakpoint 3q27, a translocation t(14;18), or the absence of both. Group I involved a t(14;18) and no 3q27 aberrations (n = 13); group II was without a t(14;18) and without 3q27 aberrations (n = 9), but had other cytogenetic aberrations; and group III was without a t(14;18) but with aberrations involving 3q27 (n = 10). None of the FL grade 3B cases harbored both a t(14;18) and 3q27 aberration. These results, in particular the finding of a mutual exclusiveness of bcl-2 and bcl-6 rearrangement, indicate at least 3 different pathways of oncogenesis in FL grade 3B. FL grade 3B with bcl-2 rearrangement probably is part of the same entity as the other follicular lymphomas (1, 2, 3A), whereas the cases with 3q27 abnormalities or other unrelated translocations are more closely related to the majority of diffuse large-cell lymphomas of germinal center cell origin. PMID- 12529294 TI - Decreased phosphorylation of protein kinase B and extracellular signal-regulated kinase in neutrophils from patients with myelodysplasia. AB - Neutrophils from patients with myelodysplastic syndrome (MDS) show a disturbed differentiation pattern and are generally dysfunctional. To study these defects in more detail, we investigated reactive-oxygen species (ROS) production and F actin polymerization in neutrophils from MDS patients and healthy controls and the involvement of N-formyl-L-methionyl-L-lucyl-L-phenylaline (fMLP) and granulocyte macrophage-colony-stimulating factor (GM-CSF)-stimulated signal transduction pathways. Following fMLP stimulation, similar levels of respiratory burst, F-actin polymerization, and activation of the small GTPase Rac2 were demonstrated in MDS and normal neutrophils. However, GM-CSF and G-CSF priming of ROS production were significantly decreased in MDS patients. We subsequently investigated the signal transduction pathways involved in ROS generation and demonstrated that fMLP-stimulated ROS production was inhibited by the phosphatidylinositol 3 kinase (PI3K) inhibitor LY294002, but not by the MAPK/ERK kinase (MEK) inhibitor U0126. In contrast, ROS production induced by fMLP stimulation of GM-CSF-primed cells was inhibited by LY294002 and U0126. This coincides with enhanced protein kinase B (PKB/Akt) phosphorylation that was PI3K dependent and enhanced extracellular signal-regulated protein kinase 1 and 2 (ERK1/2) phosphorylation that was PI3K independent. We demonstrated higher protein levels of the PI3K subunit p110 in neutrophils from MDS patients and found that though the fMLP-induced phosphorylation of PKB/Akt and ERK1/2 could also be enhanced by pretreatment with GM-CSF in these patients, the degree and kinetics of PKB/Akt and ERK1/2 phosphorylation were significantly disturbed. These defects were observed despite a normal GM-CSF-induced signal transducer and activator of transcription 5 (STAT5) phosphorylation. Our results indicate that the reduced priming of neutrophil ROS production in MDS patients might be caused by a disturbed convergence of the fMLP and GM-CSF signaling routes. PMID- 12529295 TI - Molecular response to imatinib mesylate following relapse after allogeneic SCT for CML. PMID- 12529296 TI - Graft versus myeloma may overcome the unfavorable effect of deletion of chromosome 13 in multiple myeloma. PMID- 12529297 TI - Not every polymorphism close to the AUG codon can be explained by invoking context effects on initiation of translation. PMID- 12529298 TI - Polymorphic expression of CD158k/p140/KIR3DL2 in Sezary patients. PMID- 12529299 TI - Differences between the N-glycans of human serum erythropoietin and recombinant human erythropoietin. PMID- 12529300 TI - Should patients with FMF undergo BMT? PMID- 12529301 TI - Cross-species sequence comparisons: a review of methods and available resources. AB - With the availability of whole-genome sequences for an increasing number of species, we are now faced with the challenge of decoding the information contained within these DNA sequences. Comparative analysis of DNA sequences from multiple species at varying evolutionary distances is a powerful approach for identifying coding and functional noncoding sequences, as well as sequences that are unique for a given organism. In this review, we outline the strategy for choosing DNA sequences from different species for comparative analyses and describe the methods used and the resources publicly available for these studies. PMID- 12529302 TI - Covariation in frequencies of substitution, deletion, transposition, and recombination during eutherian evolution. AB - Six measures of evolutionary change in the human genome were studied, three derived from the aligned human and mouse genomes in conjunction with the Mouse Genome Sequencing Consortium, consisting of (1) nucleotide substitution per fourfold degenerate site in coding regions, (2) nucleotide substitution per site in relics of transposable elements active only before the human-mouse speciation, and (3) the nonaligning fraction of human DNA that is nonrepetitive or in ancestral repeats; and three derived from human genome data alone, consisting of (4) SNP density, (5) frequency of insertion of transposable elements, and (6) rate of recombination. Features 1 and 2 are measures of nucleotide substitutions at two classes of "neutral" sites, whereas 4 is a measure of recent mutations. Feature 3 is a measure dominated by deletions in mouse, whereas 5 represents insertions in human. It was found that all six vary significantly in megabase sized regions genome-wide, and many vary together. This indicates that some regions of a genome change slowly by all processes that alter DNA, and others change faster. Regional variation in all processes is correlated with, but not completely accounted for, by GC content in human and the difference between GC content in human and mouse. PMID- 12529304 TI - Genome rearrangements in mammalian evolution: lessons from human and mouse genomes. AB - Although analysis of genome rearrangements was pioneered by Dobzhansky and Sturtevant 65 years ago, we still know very little about the rearrangement events that produced the existing varieties of genomic architectures. The genomic sequences of human and mouse provide evidence for a larger number of rearrangements than previously thought and shed some light on previously unknown features of mammalian evolution. In particular, they reveal that a large number of microrearrangements is required to explain the differences in draft human and mouse sequences. Here we describe a new algorithm for constructing synteny blocks, study arrangements of synteny blocks in human and mouse, derive a most parsimonious human-mouse rearrangement scenario, and provide evidence that intrachromosomal rearrangements are more frequent than interchromosomal rearrangements. Our analysis is based on the human-mouse breakpoint graph, which reveals related breakpoints and allows one to find a most parsimonious scenario. Because these graphs provide important insights into rearrangement scenarios, we introduce a new visualization tool that allows one to view breakpoint graphs superimposed with genomic dot-plots. PMID- 12529305 TI - Leveraging the mouse genome for gene prediction in human: from whole-genome shotgun reads to a global synteny map. AB - The availability of draft sequences for both the mouse and human genomes makes it possible, for the first time, to annotate whole mammalian genomes using comparative methods. TWINSCAN is a gene-prediction system that combines the methods of single-genome predictors like GENSCAN with information derived from genome comparison, thereby improving accuracy. Because TWINSCAN uses genomic sequence only, it is less biased toward highly and/or ubiquitously expressed genes than GENEWISE, GENOMESCAN, and other methods based on evidence derived from transcripts. We show that TWINSCAN improves gene prediction in human using intermediate products from various stages of the sequencing and analysis of the mouse genome, from low-redundancy, whole-genome shotgun reads to the draft assembly and the synteny map. TWINSCAN improves on the prior state of the art even when alignments from only 1X coverage of the mouse genome are available. Gene prediction accuracy improves steadily from 1X through 3X, more slowly from 3X to 4X, and relatively little thereafter. The assembly and the synteny map greatly speed the computations, however. Our human annotation using the mouse assembly is conservative, predicting only 25,622 genes, and appears to be one of the best de novo annotations of the human genome to date. PMID- 12529306 TI - Pericentromeric duplications in the laboratory mouse. AB - Duplications have long been postulated to be an important mechanism by which genomes evolve. Interspecies genomic comparisons are one method by which the origin and molecular mechanism of duplications can be inferred. By comparative mapping in human, mouse, and rat, we previously found evidence for a recent chromosome-fission event that occurred in the mouse lineage. Cytogenetic mapping revealed that the genomic segments flanking the fission site appeared to be duplicated, with copies residing near the centromere of multiple mouse chromosomes. Here we report the mapping and sequencing of the regions of mouse chromosomes 5 and 6 involved in this chromosome-fission event as well as the results of comparative sequence analysis with the orthologous human and rat genomic regions. Our data indicate that the duplications associated with mouse chromosomes 5 and 6 are recent and that the resulting duplicated segments share significant sequence similarity with a series of regions near the centromeres of the mouse chromosomes previously identified by cytogenetic mapping. We also identified pericentromeric duplicated segments shared between mouse chromosomes 5 and 1. Finally, novel mouse satellite sequences as well as putative chimeric transcripts were found to be associated with the duplicated segments. Together, these findings demonstrate that pericentromeric duplications are not restricted to primates and may be a common mechanism for genome evolution in mammals. PMID- 12529307 TI - Distinguishing regulatory DNA from neutral sites. AB - We explore several computational approaches to analyzing interspecies genomic sequence alignments, aiming to distinguish regulatory regions from neutrally evolving DNA. Human-mouse genomic alignments were collected for three sets of human regions: (1) experimentally defined gene regulatory regions, (2) well characterized exons (coding sequences, as a positive control), and (3) interspersed repeats thought to have inserted before the human-mouse split (a good model for neutrally evolving DNA). Models that potentially could distinguish functional noncoding sequences from neutral DNA were evaluated on these three data sets, as well as bulk genome alignments. Our analyses show that discrimination based on frequencies of individual nucleotide pairs or gaps (i.e., of possible alignment columns) is only partially successful. In contrast, scoring procedures that include the alignment context, based on frequencies of short runs of alignment columns, dramatically improve separation between regulatory and neutral features. Such scoring functions should aid in the identification of putative regulatory regions throughout the human genome. PMID- 12529308 TI - Strategies and tools for whole-genome alignments. AB - The availability of the assembled mouse genome makes possible, for the first time, an alignment and comparison of two large vertebrate genomes. We investigated different strategies of alignment for the subsequent analysis of conservation of genomes that are effective for assemblies of different quality. These strategies were applied to the comparison of the working draft of the human genome with the Mouse Genome Sequencing Consortium assembly, as well as other intermediate mouse assemblies. Our methods are fast and the resulting alignments exhibit a high degree of sensitivity, covering more than 90% of known coding exons in the human genome. We obtained such coverage while preserving specificity. With a view towards the end user, we developed a suite of tools and Web sites for automatically aligning and subsequently browsing and working with whole-genome comparisons. We describe the use of these tools to identify conserved non-coding regions between the human and mouse genomes, some of which have not been identified by other methods. PMID- 12529303 TI - Reevaluating human gene annotation: a second-generation analysis of chromosome 22. AB - We report a second-generation gene annotation of human chromosome 22. Using expressed sequence databases, comparative sequence analysis, and experimental verification, we have extended genes, fused previously fragmented structures, and identified new genes. The total length in exons of annotation was increased by 74% over our previously published annotation and includes 546 protein-coding genes and 234 pseudogenes. Thirty-two potential protein-coding annotations are partial copies of other genes, and may represent duplications on an evolutionary path to change or loss of function. We also identified 31 non-protein-coding transcripts, including 16 possible antisense RNAs. By extrapolation, we estimate the human genome contains 29,000-36,000 protein-coding genes, 21,300 pseudogenes, and 1500 antisense RNAs. We suggest that our revised annotation criteria provide a paradigm for future annotation of the human genome. PMID- 12529309 TI - The phusion assembler. AB - The Phusion assembler has assembled the mouse genome from the whole-genome shotgun (WGS) dataset collected by the Mouse Genome Sequencing Consortium, at ~7.5x sequence coverage, producing a high-quality draft assembly 2.6 gigabases in size, of which 90% of these bases are in 479 scaffolds. For the mouse genome, which is a large and repeat-rich genome, the input dataset was designed to include a high proportion of paired end sequences of various size selected inserts, from 2-200 kbp lengths, into various host vector templates. Phusion uses sequence data, called reads, and information about reads that share common templates, called read pairs, to drive the assembly of this large genome to highly accurate results. The preassembly stage, which clusters the reads into sensible groups, is a key element of the entire assembler, because it permits a simple approach to parallelization of the assembly stage, as each cluster can be treated independent of the others. In addition to the application of Phusion to the mouse genome, we will also present results from the WGS assembly of Caenorhabditis briggsae sequenced to about 11x coverage. The C. briggsae assembly was accessioned through EMBL, http://www.ebi.ac.uk/services/index.html, using the series CAAC01000001-CAAC01000578, however, the Phusion mouse assembly described here was not accessioned. The mouse data was generated by the Mouse Genome Sequencing Consortium. The C. briggsae sequence was generated at The Wellcome Trust Sanger Institute and the Genome Sequencing Center, Washington University School of Medicine. PMID- 12529310 TI - Whole-genome sequence assembly for mammalian genomes: Arachne 2. AB - We previously described the whole-genome assembly program Arachne, presenting assemblies of simulated data for small to mid-sized genomes. Here we describe algorithmic adaptations to the program, allowing for assembly of mammalian-size genomes, and also improving the assembly of smaller genomes. Three principal changes were simultaneously made and applied to the assembly of the mouse genome, during a six-month period of development: (1) Supercontigs (scaffolds) were iteratively broken and rejoined using several criteria, yielding a 64-fold increase in length (N50), and apparent elimination of all global misjoins; (2) gaps between contigs in supercontigs were filled (partially or completely) by insertion of reads, as suggested by pairing within the supercontig, increasing the N50 contig length by 50%; (3) memory usage was reduced fourfold. The outcome of this mouse assembly and its analysis are described in (Mouse Genome Sequencing Consortium 2002). PMID- 12529311 TI - AVID: A global alignment program. AB - In this paper we describe a new global alignment method called AVID. The method is designed to be fast, memory efficient, and practical for sequence alignments of large genomic regions up to megabases long. We present numerous applications of the method, ranging from the comparison of assemblies to alignment of large syntenic genomic regions and whole genome human/mouse alignments. We have also performed a quantitative comparison of AVID with other popular alignment tools. To this end, we have established a format for the representation of alignments and methods for their comparison. These formats and methods should be useful for future studies. The tools we have developed for the alignment comparisons, as well as the AVID program, are publicly available. See Web Site References section for AVID Web address and Web addresses for other programs discussed in this paper. PMID- 12529312 TI - Human-mouse alignments with BLASTZ. AB - The Mouse Genome Analysis Consortium aligned the human and mouse genome sequences for a variety of purposes, using alignment programs that suited the various needs. For investigating issues regarding genome evolution, a particularly sensitive method was needed to permit alignment of a large proportion of the neutrally evolving regions. We selected a program called BLASTZ, an independent implementation of the Gapped BLAST algorithm specifically designed for aligning two long genomic sequences. BLASTZ was subsequently modified, both to attain efficiency adequate for aligning entire mammalian genomes and to increase its sensitivity. This work describes BLASTZ, its modifications, the hardware environment on which we run it, and several empirical studies to validate its results. PMID- 12529313 TI - Comparative gene prediction in human and mouse. AB - The completion of the sequencing of the mouse genome promises to help predict human genes with greater accuracy. While current ab initio gene prediction programs are remarkably sensitive (i.e., they predict at least a fragment of most genes), their specificity is often low, predicting a large number of false positive genes in the human genome. Sequence conservation at the protein level with the mouse genome can help eliminate some of those false positives. Here we describe SGP2, a gene prediction program that combines ab initio gene prediction with TBLASTX searches between two genome sequences to provide both sensitive and specific gene predictions. The accuracy of SGP2 when used to predict genes by comparing the human and mouse genomes is assessed on a number of data sets, including single-gene data sets, the highly curated human chromosome 22 predictions, and entire genome predictions from ENSEMBL. Results indicate that SGP2 outperforms purely ab initio gene prediction methods. Results also indicate that SGP2 works about as well with 3x shotgun data as it does with fully assembled genomes. SGP2 provides a high enough specificity that its predictions can be experimentally verified at a reasonable cost. SGP2 was used to generate a complete set of gene predictions on both the human and mouse by comparing the genomes of these two species. Our results suggest that another few thousand human and mouse genes currently not in ENSEMBL are worth verifying experimentally. PMID- 12529314 TI - Multiple cross and inbred strain haplotype mapping of complex-trait candidate genes. AB - Identifying complex-trait candidate genes after initial low-resolution mapping has proven to be a difficult and labor-intensive undertaking, usually requiring years to develop and analyze congenic strains. As a result, to date, few complex trait genes have been discovered. Recently it was suggested that SNP haplotype analysis in inbred strains might be useful for mapping of complex traits. In this study, we have combined medium-resolution haplotype mapping with multiple experimental cross-mapping experiments to reduce the number of potential candidate genes in a complex-trait candidate interval. Coincident mapping of a modifier gene in multiple experimental crosses using different inbred strains is consistent with the common inheritance of a modifier allele. A haplotype map was developed in four inbred strains of mice used in our complex-trait mapping crosses across the proximal 10 cM of proximal Chromosome 19 to identify haplotype blocks that segregate appropriately. Only ~23 out of >400 genes met this criteria. This strategy coupled with tissue and expression arrays, as well as our recently described common pathway analysis to reduce the number of high-priority candidates, may provide a rapid, efficient method to identify and prioritize complex-trait candidate genes without requiring construction of congenic mouse strains. PMID- 12529315 TI - The comprehensive mouse radiation hybrid map densely cross-referenced to the recombination map: a tool to support the sequence assemblies. AB - We have developed a unique comprehensive mouse radiation hybrid (RH) map of nearly 23,000 markers integrating data from three international genome centers and over 400 independent laboratories. We have cross-referenced this map to the 0.5-cM resolution recombination-based Jackson Laboratory (TJL) backcross panel map, building a complete set of RH framework chromosome maps based on a high density of known-ordered anchor markers. We have systematically typed markers to improve coverage and resolve discrepancies, and have reanalyzed data sets as needed. The cross-linking of the RH and recombination maps has resulted in a highly accurate genome-wide map with consistent marker order. We have compared these linked framework maps to the Ensemble mouse genome sequence assembly, and show that they are a useful medium resolution tool for both validating sequence assembly and elucidating chromosome biology. PMID- 12529316 TI - The dynamics of formation and action of the ternary complex revealed in living cells using a G-protein-gated K+ channel as a biosensor. AB - Traditionally the consequences of activation of G-protein-coupled receptors (GPCRs) by an agonist are studied using biochemical assays. In this study we use live cells and take advantage of a G-protein-gated inwardly rectifying potassium channel (Kir3.1+3.2A) that is activated by the direct binding of Gbetagamma subunit to the channel complex to report, in real-time, using the patch clamp technique the activity of the "ternary complex" of agonist/receptor/G-protein. This analysis is further facilitated by the use of pertussis toxin-resistant fluorescent and non-fluorescent Galpha(i/o) subunits and a series of HEK293 cell lines stably expressing both channel and receptors (including the adenosine A(1) receptor, the adrenergic alpha(2A) receptor, the dopamine D(2S) receptor, the M4 muscarinic receptor, and the dimeric GABA-B(1b/2) receptor). We systematically analyzed the contribution of the various inputs to the observed kinetic response of channel activation. Our studies indicate that the combination of agonist, GPCR, and G-protein isoform uniquely specify the behavior of these channels and thus support the importance of the whole ternary complex at a kinetic level. PMID- 12529317 TI - Analysis of the interaction between the global regulator Mlc and EIIBGlc of the glucose-specific phosphotransferase system in Escherichia coli. AB - Mlc is a global regulator acting as a transcriptional repressor for several genes and operons of Escherichia coli encoding sugar-metabolizing enzymes and uptake systems. The repressing activity of Mlc is inactivated by binding to the dephosphorylated form of EIICB(Glc) (PtsG), which is formed during the transport of glucose. Here, we demonstrate that EIIB(Glc), the cytoplasmic domain of PtsG, alone is sufficient to inactivate Mlc but only when EIIB(Glc) is attached to the membrane by a protein anchor, which can be unrelated to PtsG. Several EIIB(Glc) mutants, which were altered in and around the phosphorylation site (Cys-421) of EIIB(Glc), were tested for their ability to bind Mlc and to affect transcriptional repression by Mlc. The exchange of Cys-421 with serine or aspartate still allowed binding to Mlc, and in addition, derepression became constitutive, i.e. independent of phosphoenolpyruvate-dependent phosphotransferase system (PTS) phosphorylation. Mutations were made in the surface-exposed residues in the vicinity of Cys-421 and identified Arg-424 as essential for binding to Mlc. Binding of Mlc to the EIIB(Glc) constructs in membrane preparations paralleled their ability to derepress Mlc-dependent transcription in vivo. These observations demonstrate that it is not the charge change at Cys-421, produced by PTS phosphorylation, that allows Mlc binding but rather the structural change in the environment surrounding Cys-421 that the phosphorylation provokes. Native Mlc exists as a tetramer. Deleting 18 amino acids from the C-terminal removes a putative amphipathic helix and results in dimeric Mlc that is no longer able to repress. PMID- 12529318 TI - Interaction of human NAD(P)H:quinone oxidoreductase 1 (NQO1) with the tumor suppressor protein p53 in cells and cell-free systems. AB - NAD(P)H:quinone oxidoreductase 1 (NQO1) has been proposed to stabilize p53 via a redox mechanism involving oxidation of NAD(P)H as a consequence of the catalytic activity of NQO1. We report that treatment of HCT-116 human colon carcinoma cells with the NQO1 inhibitor ES936 had no effect on the levels of p53 protein. ES936 is a mechanism-based inhibitor of NQO1 that irreversibly blocks the catalytic function of the enzyme. This suggests that a redox mechanism involving NQO1 mediated NAD(P)H oxidation is not responsible for the stabilization of p53. We also examined the ability of the NQO1 protein to associate with p53 using co immunoprecipitation experiments. Results from these experiments demonstrated co immunoprecipitation of NQO1 with p53 and vice versa. The association between p53 and NQO1 was not affected by treatment of HCT-116 cells with ES936, demonstrating that the association was not dependent on the catalytic activity of NQO1. A comparison of isogenic HCT-116 p53+/+ and HCT-116 p53-/- cells demonstrated an interaction of NQO1 and p53 only in the p53+/+ cells. Experiments performed in an in vitro transcription/translation system utilizing rabbit reticulocyte lysates confirmed the interaction of NQO1 and p53. In these experiments a full-length p53 coding region was used to express p53 in the presence of recombinant NQO1 protein. An association of p53 and NQO1 was also observed in primary human keratinocytes and mammary epithelial cells. In studies where mdm-2 co immunoprecipitated with p53, no association of mdm-2 with NQO1 was observed. These data demonstrate an association between p53 and NQO1 that may represent an alternate mechanism of p53 stabilization by NQO1 in a wide variety of human cell types. PMID- 12529319 TI - Mechanism of chaperone function in small heat-shock proteins. Phosphorylation induced activation of two-mode binding in alphaB-crystallin. AB - The consequences of alphaB-crystallin phosphorylation on its chaperone activity were investigated using a detailed analysis of the recognition and binding of destabilized T4 lysozyme (T4L) mutants by alphaB-crystallin phosphorylation mimics containing combinations of serine to aspartate substitutions. The T4L site directed mutants were selected to constitute an energetic ladder of progressively destabilized proteins having similar structures in the folded state. alphaB crystallin and its variants differentially recognize the T4L mutants, binding the more destabilized ones to a larger extent. Furthermore, the aspartate substitutions result in an increase in the extent of binding to the same T4L mutant and in the appearance of biphasic binding isotherms. The latter indicates the presence of two modes of binding characterized by different affinities and different numbers of binding sites. The transition to two-mode binding can also be induced by temperature or pH activation of the second mode. The similarity between the phosphorylation, pH, and temperature effects suggests a common structural origin. The location of the phosphorylation sites in the N-terminal domain and the hypothesized burial of this domain in the core of the oligomeric structure are consistent with a critical role for the destabilization of the quaternary structure in the process of recognition and binding by small heat shock proteins. PMID- 12529320 TI - The fourth epidermal growth factor-like domain of thrombomodulin interacts with the basic exosite of protein C. AB - Thrombomodulin (TM) functions as a cofactor to enhance the rate of protein C activation by thrombin approximately 1000-fold. The molecular mechanism by which TM improves the catalytic efficiency of thrombin toward protein C is not known. Molecular modeling of the protein C activation based on the crystal structure of thrombin in complex with the epidermal growth factor-like domains 4, 5, and 6 of TM (TM456) predicts that the binding of TM56 to exosite 1 of thrombin positions TM4 so that a negatively charged region on this domain juxtaposes a positively charged region of protein C. It has been hypothesized that electrostatic interactions between these oppositely charged residues of TM4 and protein C facilitate a proper docking of the substrate into the catalytic pocket of thrombin. To test this hypothesis, we have constructed several mutants of TM456 and protein C in which charges of the putative interacting residues on both TM4 (Asp/Glu) and protein C (Lys/Arg) have been reversed. Results of TM-dependent protein C activation studies by such a compensatory mutagenesis approach support the molecular model that TM4 interacts with the basic exosite of protein C. PMID- 12529321 TI - Distinct phosphoinositide 3-kinases mediate mast cell degranulation in response to G-protein-coupled versus FcepsilonRI receptors. AB - Phosphoinositide (PI) 3-kinases are critical regulators of mast cell degranulation. The Class IA PI 3-kinases p85/p110beta and p85/p110delta but not p85/p110alpha are required for antigen-mediated calcium flux in RBL-2H3 cells (Smith, A. J., Surviladze, Z., Gaudet, E. A., Backer, J. M., Mitchell, C. A., and Wilson, B. S. et al., (2001) J. Biol. Chem. 276, 17213-17220). We now examine the role of Class IA PI 3-kinases isoforms in degranulation itself, using a single cell degranulation assay that measures the binding of fluorescently tagged annexin V to phosphatidylserine in the outer leaflet of the plasma membrane of degranulated mast cells. Consistent with previous data, antibodies against p110delta and p110beta blocked FcepsilonR1-mediated degranulation in response to FcepsilonRI ligation. However, antigen-stimulated degranulation was also inhibited by antibodies against p110alpha, despite the fact that these antibodies have no effect on antigen-induced calcium flux. These data suggest that p110alpha mediates a calcium-independent signal during degranulation. In contrast, only p110beta was required for enhancement of antigen-stimulated degranulation by adenosine, which augments mast cell-mediated airway inflammation in asthma. Finally, we examined carbachol-stimulated degranulation in RBL2H3 cells stably expressing the M1 muscarinic receptor (RBL-2H3-M1 cells). Surprisingly, carbachol stimulated degranulation was blocked by antibody-mediated inhibition of the Class III PI 3-kinase hVPS34 or by titration of its product with FYVE domains. Antibodies against Class IA PI 3-kinases had no effect. These data demonstrate: (a) a calcium-independent role for p110alpha in antigen-stimulated degranulation; (b) a requirement for p110beta in adenosine receptor signaling; and (c) a requirement for hVPS34 during M1 muscarinic receptor signaling. Elucidation of the intersections between these distinct pathways will lead to new insights into mast cell degranulation. PMID- 12529323 TI - The yeast Epsin Ent1 is recruited to membranes through multiple independent interactions. AB - In addition to its well known role in targeting proteins for proteasomal degradation, ubiquitin (Ub) is also involved in promoting internalization of cell surface proteins into the endocytic pathway. Moreover, putative Ub interaction motifs (UIMs) as well as Ub-associated (UBA) domains have been identified in key yeast endocytic proteins (the epsins Ent1 and Ent2, and the Eps15 homolog Ede1). In this study, we characterized the interaction of Ub with the Ede1 UBA domain and with the UIMs of Ent1. Our data suggest that the UIMs and the UBA are involved in binding these proteins to biological membranes. We also show that the Ent1 ENTH domain binds to phosphoinositides in vitro and that Ent1 NPF motifs interact with the EH domain-containing proteins Ede1 and Pan1. Our findings indicate that the ENTH domain interaction with membrane lipids cooperates with the binding of membrane-associated Ub moieties. These events may in turn favor the occurrence of other interactions, for instance EH-NPF recognition, thus stabilizing networks of low affinity binding partners at endocytic sites. PMID- 12529322 TI - Structural basis for alpha1 versus alpha2 isoform-distinct behavior of the Na,K ATPase. AB - We showed earlier that the kinetic behavior of the alpha2 isoform of the Na,K ATPase differs from the ubiquitous alpha1 isoform primarily by a shift in the steady-state E(1)/E(2) equilibrium of alpha2 in favor of E(1) form(s). The aim of the present study was to identify regions of the alpha chain that confer the alpha1/alpha2 distinct behavior using a mutagenesis and chimera approach. Criteria to assess shifts in conformational equilibrium included (i) K(+) sensitivity of Na-ATPase measured at micromolar ATP, under which condition E(2)(K(+)) --> E(1) + K(+) becomes rate-limiting, (ii) changes in K'(ATP) for low affinity ATP binding, (iii) vanadate sensitivity of Na,K-ATPase activity, and (iv) the rate of the partial reaction E(1)P --> E(2)P. We first confirmed that interactions between the cytoplasmic domains of alpha2 that modulate conformational shifts are fundamentally similar to those of alpha1, suggesting that the predilection of alpha2 for E(1) state(s) is due to differences in primary structure of the two isoforms. Kinetic behavior of the alpha1/alpha2 chimeras indicates that the difference in E(1)/E(2) poise of the two isoforms cannot be accounted for by their notably distinct N termini, but rather by the front segment extending from the cytoplasmic N terminus to the C-terminal end of the extracellular loop between transmembranes 3 and 4, with a lesser contribution of the alpha1/alpha2 divergent portion within the M4-M5 loop near the ATP binding domain. In addition, we show that the E(1) shift of alpha2 results primarily from differences in the conformational transition of the dephosphoenzyme, (E(2)(K(+)) -> E(1) + K(+)), rather than phosphoenzyme (E(1)P --> E(2)P). PMID- 12529324 TI - Cellular prion protein sensitizes neurons to apoptotic stimuli through Mdm2 regulated and p53-dependent caspase 3-like activation. AB - We examined the influence of cellular prion protein (PrP(c)) in the control of cell death in stably transfected TSM1 cells. PrP(c) expression enhanced staurosporine-stimulated neuronal toxicity and DNA fragmentation, caspase 3-like activity and immunoreactivity, and p53 immunoreactivity and transcriptional activities. Caspase activation was reduced by the chemical inhibitor of p53, pifithrin-alpha, as well as by PrP(c)- or p53-antisense approaches but remained insensitive to the Fyn kinase inhibitor PP2 (4-amino-5-(4-chloro-phenyl)-7-(t butyl)pyrazolo[3,4-d]pyrimidine). We establish that PrP(c) controls p53 at a post transcriptional level and is reversed by Mdm2 transfection and p38 MAPK inhibitor. We propose that endogenous cellular prion protein sensitizes neurons to apoptotic stimuli through a p53-dependent caspase 3-mediated activation controlled by Mdm2 and p38 MAPK. PMID- 12529325 TI - Supplying copper to the cuproenzyme peptidylglycine alpha-amidating monooxygenase. AB - We explored the role of known copper transporters and chaperones in delivering copper to peptidylglycine-alpha-hydroxylating monooxygenase (PHM), a copper dependent enzyme that functions in the secretory pathway lumen. We examined the roles of yeast Ccc2, a P-type ATPase related to human ATP7A (Menkes disease protein) and ATP7B (Wilson disease protein), as well as yeast Atx1, a cytosolic copper chaperone. We expressed soluble PHMcc (catalytic core) in yeast using the yeast pre-pro-alpha-mating factor leader region to target the enzyme to the secretory pathway. Although the yeast genome encodes no PHM-like enzyme, PHMcc expressed in yeast is at least as active as PHMcc produced by mammalian cells. PHMcc partially co-migrated with a Golgi marker during subcellular fractionation and partially co-localized with Ccc2 based on immunofluorescence. To determine whether production of active PHM was dependent on copper trafficking pathways involving the CCC2 or ATX1 genes, we expressed PHMcc in wild-type, ccc2, and atx1 mutant yeast. Although ccc2 and atx1 mutant yeast produce normal levels of PHMcc protein, it lacks catalytic activity. Addition of exogenous copper yields fully active PHMcc. Similarly, production of active PHM in mouse fibroblasts is impaired in the presence of a mutant ATP7A gene. Although delivery of copper to lumenal cuproproteins like PAM involves ATP7A, lumenal chaperones may not be required. PMID- 12529326 TI - A large scale genetic analysis of c-Myc-regulated gene expression patterns. AB - The myc proto-oncogenes encode transcriptional regulators whose inappropriate expression is correlated with a wide array of human malignancies. Up-regulation of Myc enforces growth, antagonizes cell cycle withdrawal and differentiation, and in some situations promotes apoptosis. How these phenotypes are elicited is not well understood, largely because we lack a clear picture of the biologically relevant downstream effectors. We created a new biological system for the optimal profiling of Myc target genes based on a set of isogenic c-myc knockout and conditional cell lines. The ability to modulate Myc activity from essentially null to supraphysiological resulted in a significantly increased and reproducible yield of targets and revealed a large subset of genes that respond optimally to Myc in its physiological range of expression. The total extent of transcriptional changes that can be triggered by Myc is remarkable and involves thousands of genes. Although the majority of these effects are not direct, many of the indirect targets are likely to have important roles in mediating the elicited cellular phenotypes. Myc-activated functions are indicative of a physiological state geared toward the rapid utilization of carbon sources, the biosynthesis of precursors for macromolecular synthesis, and the accumulation of cellular mass. In contrast, the majority of Myc-repressed genes are involved in the interaction and communication of cells with their external environment, and several are known to possess antiproliferative or antimetastatic properties. PMID- 12529327 TI - The tetrameric structure of Haemophilus influenza hybrid Prx5 reveals interactions between electron donor and acceptor proteins. AB - Cellular redox control is often mediated by oxidation and reduction of cysteine residues in the redox-sensitive proteins, where thioredoxin and glutaredoxin (Grx) play as electron donors for the oxidized proteins. Despite the importance of protein-protein interactions between the electron donor and acceptor proteins, there has been no structural information for the interaction of thioredoxin or Grx with natural target proteins. Here, we present the crystal structure of a novel Haemophilus influenza peroxiredoxin (Prx) hybrid Prx5 determined at 2.8-A resolution. The structure reveals that hybrid Prx5 forms a tightly associated tetramer where active sites of Prx and Grx domains of different monomers interact with each other. The Prx-Grx interface comprises specific charge interactions surrounded by weak interactions, providing insight into the target recognition mechanism of Grx. The tetrameric structure also exhibits a flexible active site and alternative Prx-Grx interactions, which appear to facilitate the electron transfer from Grx to Prx domain. Differences of electron donor binding surfaces in Prx proteins revealed by an analysis based on the structural information explain the electron donor specificities of various Prx proteins. PMID- 12529328 TI - MAP kinase-dependent degradation of p27Kip1 by calpains in choroidal melanoma cells. Requirement of p27Kip1 nuclear export. AB - We investigated the status and the regulation of the cyclin-dependent kinases (CDK) inhibitor p27(Kip1) in a choroidal melanoma tumor-derived cell line (OCM 1). By contrast to normal choroidal melanocytes, the expression level of p27(Kip1) was low in these cells and the mitogen-activated protein (MAP) kinase pathway was constitutively activated. Genetic or chemical inhibition of this pathway induced p27(Kip1) accumulation, whereas MAP kinase reactivation triggered a down-regulation of p27(Kip1) that could be partially reversed by calpain inhibitors. In good accordance, ectopic expression of the cellular calpain inhibitor calpastatin led to an increase of endogenous p27(Kip1) expression. In vitro, p27(Kip1) was degraded by calpains, and OCM-1 cell extracts contained a calcium-dependent p27(Kip1) degradation activity. MAP kinase inhibition partially inhibited both calpain activity and calcium-dependent p27(Kip1) degradation by cellular extracts. Immunofluorescence labeling and subcellular fractionation revealed that p27(Kip1) was in part localized in the cytoplasmic compartment of OCM-1 cells but not of melanocytes, and accumulated into the nucleus upon MAP kinase inhibition. MAP kinase activation triggered a cytoplasmic translocation of the protein, as well as a change in its phosphorylation status. This CRM-1 dependent cytoplasmic translocation was necessary for MAP kinase- and calpain dependent degradation. Taken together, these data suggest that in tumor-derived cells, p27(Kip1) could be degraded by calpains through a MAP kinase-dependent process, and that abnormal cytoplasmic localization of the protein, probably linked to modifications of its phosphorylation state, could be involved in this alternative mechanism of degradation. PMID- 12529329 TI - Crystal structure of a novel regulatory 40-kDa mammary gland protein (MGP-40) secreted during involution. AB - We have determined the crystal structure of a novel regulatory protein (MGP-40) from the mammary gland. This protein is implicated as a protective signaling factor that determines which cells are to survive the drastic tissue remodeling that occurs during involution. It has been indicated that certain cancers could surreptitiously utilize the proposed normal protective signaling by proteins of this family to extend their own survival and thereby allow them to invade the organ and metastasize. In view of this, MGP-40 could form an important target for rational structure-based drug design against breast cancer. It is a single chain, glycosylated protein with a molecular mass of 40 kDa. It was isolated from goat dry secretions and has been cloned and sequenced. It was crystallized by microdialysis from 20 mg ml(-1) solution in 0.1 m Tris-HCl, pH 8.0, and equilibrated against the same solution containing 19% ethanol. Its x-ray structure has been determined by molecular replacement and refined to a 2.9 A resolution. The protein adopts a beta/alpha domain structure with a triose phosphate isomerase barrel conformation in the core and a small alpha+beta folding domain. A single glycosylation site containing two N-acetylglucosamine units has been observed in the structure. Compared with chitinases and chitinase like proteins the most important mutation in this protein pertains to a change from Glu to Leu at position 119, which is part of the so-called active site sequence in the form of Asp(115), Leu(119), and Asp(186) and in this case resulting in the loss of chitinase activity. The orientations of two Trp residues Trp(78) and Trp(331) in the beta barrel reduces the free space, drastically impairing the binding of saccharides/polysaccharides. However, the site and mode of binding of this protein to cell surface receptors are not yet known. PMID- 12529330 TI - Arsenite-induced phosphorylation of histone H3 at serine 10 is mediated by Akt1, extracellular signal-regulated kinase 2, and p90 ribosomal S6 kinase 2 but not mitogen- and stress-activated protein kinase 1. AB - Arsenite is known to be an environmental human carcinogen. However, the mechanism of action of this compound in skin carcinogenesis is not completely clear. Here, we provide evidence that arsenite can induce phosphorylation of histone H3 at serine 10 in a time- and dose-dependent manner in JB6 Cl 41 cells. Arsenite induces phosphorylation of Akt1 at serine 473 and increases Akt1 activity. A dominant-negative mutant of Akt1 inhibits the arsenite-induced phosphorylation of histone H3 at serine 10. Additionally, active Akt1 kinase strongly phosphorylates histone H3 at serine 10 in vitro. The arsenite-induced phosphorylation of histone H3 at serine 10 was almost completely blocked by a dominant-negative mutant of extracellular signal-regulated kinase 2 and the mitogen-activated protein kinase/extracellular signal-regulated kinase inhibitor PD98059. N- or C-terminal mutant mitogen- and stress-activated protein kinase 1 or its inhibitor H89 had no effect on arsenite-induced phosphorylation of histone H3 at serine 10 in JB6 Cl 41 cells. However, cells deficient in p90 ribosomal S6 kinase 2 (Rsk2(-/-)) totally block this phosphorylation in a dose- and time-dependent manner. Taken together, these results suggested that arsenite-induced phosphorylation of histone H3 at serine 10 is mediated by Akt1, extracellular signal-regulated kinase 2 and p90 ribosomal S6 kinase 2 but not mitogen- and stress-activated protein kinase 1. PMID- 12529331 TI - A general strategy to uncover transcription factor properties identifies a new regulator of drug resistance in yeast. AB - We demonstrate a genomewide approach to determine the physiological role of a putative transcription factor, Ylr266, identified through yeast genome sequencing program. We constructed activated forms of the zinc finger (Zn(2)Cys(6)) protein Ylr266, and we analyzed the corresponding transcriptomes with DNA microarrays to characterize the up-regulated genes. The direct target genes of Ylr266 were further identified by in vivo chromatin immunoprecipitation procedure. The functions of the genes directly controlled by YLR266c are in agreement with the observed drug-resistance phenotype of the cell expressing an activated form of Ylr266. These target genes code for ATP-binding cassette or major facilitator superfamily transporters such as PDR15, YOR1, or AZR1 or for other proteins such as SNG1, YJL216c, or YLL056c which are already known to be involved in the yeast pleiotropic drug resistance (PDR) phenomenon. YLR266c could thus be named PDR8. Overlaps with the other PDR networks argue in favor of a new specific role for PDR8 in connection with the well known PDR regulators PDR1/PDR3 and YRR1. This strategy to identify the regulatory properties of an anonymous transcription factor is likely to be generalized to all the Zn(2)Cys(6) transcription factors from Saccharomyces cerevisiae and related yeasts. PMID- 12529332 TI - Ceramide-induced and age-associated increase in macrophage COX-2 expression is mediated through up-regulation of NF-kappa B activity. AB - We have shown that the age-associated increase in lipopolysaccharide (LPS) stimulated macrophages (M phi) prostaglandin E(2) (PGE(2)) production is because of ceramide-induced up-regulation of cyclooxygenase (COX)-2 transcription that leads to increased COX-2 expression and enzyme activity. To determine the mechanism of the age-related and ceramide-dependent increase in COX-2 transcription, we investigated the role of various transcription factors involved in COX-2 gene expression. The results showed that LPS-initiated activations of both consensus and COX-2-specific NF-kappa B, but not AP-1 and CREB, were significantly higher in M phi from old mice than those from young mice. We further showed that the higher NF-kappa B activation in old M phi was because of greater I kappa B degradation in the cytoplasm and p65 translocation to the nucleus. An I kappa B phosphorylation inhibitor, Bay 11-7082, inhibited NF-kappa B activation, as well as PGE(2) production, COX activity, COX-2 protein, and mRNA expression in both young and old M phi. Similar results were obtained by blocking NF-kappa B binding activity using a NF-kappa B decoy. Furthermore, NF-kappa B inhibition resulted in significantly greater reduction in PGE(2) production and COX activity in old compared with young M phi. Addition of ceramide to the young M phi, in the presence or absence of LPS, increased NF-kappa B activation in parallel with PGE(2) production. Bay 11-7082 or NF-kappa B decoy prevented this ceramide-induced increase in NF-kappa B binding activity and PGE(2) production. These findings strongly suggest that the age-associated and ceramide-induced increase in COX-2 transcription is mediated through higher NF-kappa B activation, which is, in turn, because of a greater I kappa B degradation in old M phi. PMID- 12529333 TI - Sumoylation of the progesterone receptor and of the steroid receptor coactivator SRC-1. AB - SUMO-1 (small ubiquitin-like modifier) conjugation regulates the subcellular localization, stability, and activity of a variety of proteins. We show here that SUMO-1 overexpression markedly enhances progesterone receptor (PR)-mediated gene transcription. PR undergoes a sumoylation at lysine 388 located in its N-terminal domain. However, sumoylation of the receptor is not responsible for enhanced transcription because substitution of its target lysine did not abolish the effect of SUMO-1 and even converted the receptor into a slightly more active transactivator. Furthermore estrogen receptor alpha (ERalpha)-driven transcription is also enhanced by SUMO-1 overexpression contrasting with the absence of sumoylation of this receptor. We thus analyzed SUMO-1 conjugation to the steroid receptor coactivator SRC-1. We showed that this protein contains two major sites of conjugation at Lys-732 and Lys-774. Sumoylation was shown to increase PR-SRC-1 interaction and to prolong SRC-1 retention in the nucleus. It did not prevent SRC-1 ubiquitinylation and did not exert a clear effect on the stability of the protein. Overexpression of SUMO-1 enhanced PR-mediated gene transcription even in the presence of non-sumoylated mutants of SRC-1. This observation suggests that among the many protein partners involved in steroid hormone-mediated gene regulation several are probably targets of SUMO-1 modification. PMID- 12529334 TI - Phosphorylation of p16INK4A correlates with Cdk4 association. AB - Progression through the eukaryotic cell cycle is driven by the activity of cyclin dependent kinases. The cyclin D-dependent kinase Cdk4 promotes progression through the G(1) phase of the cell cycle and is deregulated in many human tumors. The tumor suppressor protein p16(INK4A) (p16) forms a complex with Cdk4 and inhibits kinase activity. Here we report that p16 is phosphorylated, and the phosphorylated form of p16 is preferentially associated with Cdk4 in normal human fibroblasts. We mapped phosphorylation sites on exogenously overexpressed p16 to serines 7, 8, 140, and 152 and found that endogenous p16 associated with Cdk4 is phosphorylated at serine 152. All mapped phosphorylation sites lie outside of the conserved kinase-binding domain of p16 but in regions of the protein affected by mutations in familial and sporadic cancer. Our results suggest a novel regulation of p16 activity. PMID- 12529335 TI - Preventing hormone-dependent breast cancer in high-risk women. PMID- 12529336 TI - Early-stage ovarian cancer: to treat or not to treat. PMID- 12529337 TI - Keeping up with alternative medicine: researchers offer evaluation criteria. PMID- 12529339 TI - Selenium for prevention: eating your way to better DNA repair? PMID- 12529340 TI - Estrogen and DNA damage: the silent source of breast cancer? PMID- 12529341 TI - Success of vaccine offers promise of cervical cancer prevention. PMID- 12529342 TI - Stat bite: incidence of cervical cancer in white and Hispanic women, 1992-1999. PMID- 12529343 TI - International Collaborative Ovarian Neoplasm trial 1 and Adjuvant ChemoTherapy In Ovarian Neoplasm trial: two parallel randomized phase III trials of adjuvant chemotherapy in patients with early-stage ovarian carcinoma. AB - BACKGROUND: Adjuvant chemotherapy has been suggested as a possible strategy to improve survival in women with early-stage ovarian cancer; however, all randomized studies to date have been too small to answer this question reliably. METHODS: We performed a preplanned combined analysis of two parallel randomized clinical trials (International Collaborative Ovarian Neoplasm 1 [ICON1] and Adjuvant ChemoTherapy In Ovarian Neoplasm [ACTION]) in early-stage ovarian cancer that compared platinum-based adjuvant chemotherapy with observation following surgery. Between November 1990 and January 2000, 925 patients (477 in ICON1 and 448 in ACTION) who had surgery for early-stage ovarian cancer were randomly assigned to receive platinum-based adjuvant chemotherapy (n = 465) or observation (n = 460) until chemotherapy was indicated. Kaplan-Meier analysis was used to compare overall and recurrence-free survival by treatment allocation. In subgroup analyses of pretreatment age, tumor stage, histologic cell type, and differentiation grade, the differences in relative size of effect were tested using a chi-square test for interaction or a chi-square test for trend. All tests of statistical significance were two-sided. RESULTS: After a median follow-up of over 4 years, 245 patients had died or had a recurrence (ICON1: 133, ACTION: 112). Overall survival at 5 years was 82% in the chemotherapy arm and 74% in the observation arm (difference = 8% [95% confidence interval (CI) = 2% to 12%]; hazard ratio [HR] = 0.67, 95% CI = 0.50 to 0.90; P =.008). Recurrence-free survival at 5 years was also better in the adjuvant chemotherapy arm than it was in the observation arm (76% versus 65%, difference = 11% [95% CI = 5% to 16%]; HR = 0.64, 95% CI = 0.50 to 0.82; P =.001). Subgroup analyses provided no evidence of a difference in the size of effect of chemotherapy on survival in any pretreatment subcategory. CONCLUSIONS: Platinum-based adjuvant chemotherapy improved overall survival and recurrence-free survival at 5 years in this combined group of patients with early-stage ovarian cancer defined by the inclusion criteria of the ICON1 and ACTION trials. PMID- 12529344 TI - Impact of adjuvant chemotherapy and surgical staging in early-stage ovarian carcinoma: European Organisation for Research and Treatment of Cancer-Adjuvant ChemoTherapy in Ovarian Neoplasm trial. AB - BACKGROUND: All randomized trials of adjuvant chemotherapy for early-stage ovarian cancer have lacked the statistical power to show a difference in the effect on survival between adjuvant chemotherapy and no adjuvant chemotherapy. They have also not taken into account the adequacy of surgical staging. We performed a prospective unblinded, randomized phase III trial to test the efficacy of adjuvant chemotherapy in patients with early-stage ovarian cancer, with emphasis on the extent of surgical staging. METHODS: Between November 1990 and January 2000, 448 patients from 40 centers in nine European countries were randomly assigned to either adjuvant platinum-based chemotherapy (n = 224) or observation (n = 224) following surgery. Endpoints were overall survival and recurrence-free survival, and the analysis was on an intention-to-treat basis. The Kaplan-Meier method was used to perform time-to-event analysis, and the log rank test was used to compare differences between treatment arms. Statistical tests were two-sided. RESULTS: After a median follow-up of 5.5 years, the difference in overall survival between the two trial arms was not statistically significant (hazard ratio [HR] = 0.69, 95% confidence interval [CI] = 0.44 to 1.08; P =.10). Recurrence-free survival, however, was statistically significantly improved in the adjuvant chemotherapy arm (HR = 0.63, 95% CI = 0.43 to 0.92; P =.02). Approximately one-third of patients (n = 151) had been optimally staged and two-thirds (n = 297) had not. Among patients in the observation arm, optimal staging was associated with a statistically significant improvement in overall and recurrence-free survival (HR = 2.31 [95% CI = 1.08 to 4.96]; P =.03 and HR = 1.82 [95% CI = 1.02 to 3.24] P =.04, respectively). No such association was observed in the chemotherapy arm. In the non-optimally staged patients, adjuvant chemotherapy was associated with statistically significant improvements in overall and recurrence-free survival (HR = 1.75 [95% CI = 1.04 to 2.95]; P =.03 and HR = 1.78 [95% CI = 1.15 to 2.77]; P =.009, respectively). In the optimally staged patients, no benefit of adjuvant chemotherapy was seen. CONCLUSION: Adjuvant chemotherapy was associated with statistically significantly improved recurrence-free survival in patients with early-stage ovarian cancer. The benefit of adjuvant chemotherapy appeared to be limited to patients with non-optimal staging, i.e., patients with more risk of unappreciated residual disease. PMID- 12529345 TI - International Collaborative Ovarian Neoplasm trial 1: a randomized trial of adjuvant chemotherapy in women with early-stage ovarian cancer. AB - BACKGROUND: The question of whether platinum-based adjuvant chemotherapy can improve outcomes in patients with early-stage epithelial ovarian cancer is an important one. We carried out a multicenter, open randomized trial to determine whether adjuvant chemotherapy would improve overall survival and prolong recurrence-free survival in women with early-stage epithelial ovarian cancer. METHODS: Between August 1991 and January 2000, 477 patients in 84 centers in five countries were randomly assigned to receive either adjuvant chemotherapy immediately following surgery (n = 241) or no adjuvant chemotherapy until clinically indicated (n = 236). Kaplan-Meier curves of overall survival and recurrence-free survival were compared using the Mantel-Cox version of the log rank test. All statistical tests were two-sided. RESULTS: Women who received adjuvant chemotherapy had better overall survival than women who did not (hazard ratio [HR] of 0.66, 95% confidence interval [CI] = 0.45 to 0.97; P =.03). These results translate into 5-year survival figures of 70% for women who did not receive adjuvant chemotherapy and 79% for women who did receive adjuvant chemotherapy, a difference of 9% (95% CI = 1% to 15%). Adjuvant chemotherapy also improved recurrence-free survival (HR = 0.65; 95% CI = 0.46 to 0.91; P =.01). These results translate into 5-year recurrence-free survival figures of 62% for women who did not receive adjuvant chemotherapy and 73% for women who did receive adjuvant chemotherapy, a difference of 11% (95% CI = 3% to 18%). CONCLUSION: These results suggest that platinum-based adjuvant chemotherapy improves survival and delays recurrence in patients with early-stage ovarian cancer. PMID- 12529346 TI - Diet and sex hormones in girls: findings from a randomized controlled clinical trial. AB - BACKGROUND: Results of several studies have suggested that diet during adolescence may influence the risk of breast cancer in adulthood. We evaluated whether an intervention to lower fat intake among adolescent girls altered their serum concentrations of sex hormones that, in adults, are related to breast cancer development. METHODS: We conducted an ancillary hormone study among 286 of the 301 girls who participated between 1988 and 1997 in the Dietary Intervention Study in Children, in which healthy, prepubertal, 8- to 10-year-olds with elevated low-density lipoprotein cholesterol were randomly assigned to usual care or to a behavioral intervention that promoted a low-fat diet. Median time on the intervention was 7 years. Blood samples collected before randomization and at the year 1, year 3, year 5, and last visits were assayed to determine the girls' serum levels of sex hormones. All P values are two-sided. RESULTS: At the year 5 visit, girls in the intervention group had 29.8% (95% confidence interval [CI] = 5.4% to 47.9%; P =.02) lower estradiol, 30.2% (95% CI = 7.0% to 47.7%; P =.02) lower non-sex hormone binding globulin-bound estradiol, 20.7% (95% CI = 4.7% to 34.0%; P =.02) lower estrone, and 28.7% (95% CI = 5.1% to 46.5%; P =.02) lower estrone sulfate levels during the follicular phase of the menstrual cycle and 27.2% (95% CI = 5.7% to 53.1%; P =.01) higher testosterone levels during the luteal phase of the menstrual cycle than did girls in the usual care group. At the last visit, the luteal phase progesterone level was 52.9% (95% CI = 20.0% to 72.3%) lower for girls in the intervention group than for girls in the usual care group (P =.007). CONCLUSION: Modest reductions in fat intake during puberty are associated with changes in sex hormone concentrations that are consistent with alterations in the function of the hypothalamic-pituitary-ovarian axis. Whether these changes influence breast cancer risk is currently unknown. PMID- 12529347 TI - Quantitative association between HER-2/neu and steroid hormone receptors in hormone receptor-positive primary breast cancer. AB - BACKGROUND: HER-2/neu, which encodes a receptor tyrosine kinase, is amplified and overexpressed in 20%-25% of human breast cancers. Such tumors are often resistant to hormone therapy. Despite a general inverse association between HER-2/neu amplification/overexpression and estrogen receptor (ER) and/or progesterone receptor (PR) expression, a fraction of patients are both HER-2/neu- and hormone receptor (HR)-positive. The efficacy of hormone therapy in this group is currently a matter of debate. To better understand the relationship between HER 2/neu positivity and HR expression, we analyzed HER-2/neu, ER, and PR as continuous variables in breast cancer cell lines and two cohorts of primary breast cancer patients. METHODS: HER-2/neu and ER/PR expression was analyzed by enzyme-linked immunosorbent assay (ELISA) and enzyme immunoassay (EIA), respectively, in 14 human breast cancer cell lines, some of which had been transfected with the HER-2/neu gene. For the clinical study population, HER-2/neu protein levels were assessed by ELISA (cohort A, n = 665), and HER-2/neu gene copy number was determined using fluorescence in situ hybridization (cohort B, n = 894). ER/PR expression was analyzed by EIA (cohort A) or radioligand binding (cohort B). Associations between HER-2/neu and ER/PR expression were analyzed using Spearman's rho correlation and the chi-square test, and absolute levels were compared using the Mann-Whitney U test. All statistical tests were two sided. RESULTS: HR-positive human breast cancer cell lines transfected with the HER-2/neu gene expressed statistically significantly lower levels of ER and PR than parental lines. In the clinical cohorts, levels of HER-2/neu overexpression and gene amplification were inversely correlated with ER/PR levels (Cohort A [n = 112]: for ER, r = -0.34, P<.001; for PR, r = -0.24, P =.010. Cohort B [n = 188]: for ER, r = -0.39, P<.001; for PR, r = -0.26, P<.001). Among patients with HR positive tumors, HER-2/neu-positive tumors had statistically significantly lower ER/PR levels than HER-2/neu-negative ones (Cohort A: for ER, median = 25 fmol/mg [interquartile range [IQR] = 13-78] versus median = 38.5 fmol/mg [IQR = 17-99] and P =.031; for PR, median = 35 fmol/mg [IQR = 12-119] versus median = 88.5 fmol/mg [IQR = 22-236] and P<.001. Cohort B: for ER, median = 44 fmol/mg [IQR = 13-156] versus median = 92 fmol/mg [IQR = 35-235] and P<.001; for PR, median = 36 fmol/mg [IQR = 13-108] versus median = 84 fmol/mg [IQR = 24-250] and P<.001). Patients with higher levels of HER-2/neu overexpression or amplification had statistically significantly lower levels of ER/PR than patients with lower levels of HER-2/neu overexpression or amplification. CONCLUSION: Because absolute HR levels are strongly related to response to hormone therapy in primary and advanced breast cancer, reduced ER/PR expression may be one mechanism to explain the relative resistance of HER-2/neu-positive:HR-positive tumors to hormone therapy. PMID- 12529348 TI - Association between hemochromatosis (HFE) gene mutation carrier status and the risk of colon cancer. AB - BACKGROUND: Iron is a pro-oxidant that may promote carcinogenesis. Mutations in the hemochromatosis (HFE) gene are associated with increased total body iron stores in some individuals. We assessed the risk of colon cancer among individuals with and without HFE gene mutations. METHODS: We performed a population-based, case-control study in North Carolina. Case patients with colon cancer and control subjects provided information on multiple environmental exposures, including total iron intake and nonsteroidal anti-inflammatory drug (NSAID) use. They also provided a venous blood sample, from which DNA was extracted, amplified, and subjected to diagnostic restriction enzyme mapping to detect two major HFE gene mutations, C282Y and H63D. Data were analyzed with Fisher's exact test and logistic regression. All statistical tests were two sided. RESULTS: Thirteen hundred and eight subjects participated (475 case patients, 833 control subjects). The allele frequencies of the H63D and C282Y mutations were greater among case patients (0.11 and 0.046, respectively) than among control subjects (0.09 and 0.044, respectively; P =.14 and P =.85, respectively). When we controlled for age, race, sex, red meat consumption, NSAID use, and total iron intake, subjects with any HFE gene mutation were more likely to have colon cancer than subjects with no HFE gene mutations (adjusted odds ratio [OR] = 1.40, 95% confidence interval [CI] = 1.07 to 1.87). The magnitude of the effect was similar for both the H63D (adjusted OR = 1.44, 95% CI = 1.04 to 1.98) and C282Y mutations (adjusted OR = 1.39, 95% CI = 0.88 to 2.19). The risk of colon cancer associated with an HFE gene mutation was similar for those who did and did not have a family history of colon cancer. Among those with HFE mutations, cancer risk increased with increasing age and total iron intake. CONCLUSIONS: HFE gene mutations are associated with an increased risk of colon cancer. Cancer risk is greatest in mutation carriers who are older or consume high quantities of iron. PMID- 12529349 TI - Italian randomized trial among women with hysterectomy: tamoxifen and hormone dependent breast cancer in high-risk women. AB - Tamoxifen improves outcome in women with breast cancer and reduces the incidence of estrogen receptor-positive (ER+) breast tumors in prevention trials. Tamoxifen use is associated with an increased risk of potentially serious adverse events, principally endometrial cancer and venous thromboembolic events and, therefore, detailed knowledge of the effects of tamoxifen is important. With more cases of breast cancer being found as the follow-up time increases, it is now possible to perform more detailed analysis of the Italian Randomized Trial of Tamoxifen. Women with hysterectomy (N = 5408) were randomly assigned to receive 20 mg tamoxifen per day (N = 2700) or placebo (N = 2708). After a median of 81.2 months of follow-up, 79 case subjects (34 in the tamoxifen arm and 45 in the placebo arm) were diagnosed with breast cancer. We were able to identify a group of women at increased risk of ER+ breast cancers (high-risk group) on the basis of baseline as well as reproductive and hormonal characteristics (height, age at menarche, parity, age at first birth, and oophorectomy). Tamoxifen administered to women in the high-risk group showed statistically significantly reduced incidence of breast cancer (tamoxifen, 3 and placebo, 15; P =.003), but no such effect was seen in the low-risk group (tamoxifen, 31 and placebo, 30; P =.89). The positive effect of tamoxifen on breast cancer among high-risk women is most marked for ER+ tumors (tamoxifen, 1 and placebo, 11; P =.002). Chemoprevention of breast cancer with tamoxifen appears to be effective in women at high risk of ER+ tumors but not among women at low risk, who may well be protected naturally by late age at menarche or early first pregnancy, or artificially by removal of the ovaries. Tamoxifen could be offered as a preventive agent to women identified at high-risk of breast cancer because of hormone-related risk factors. Such a strategy would greatly reduce the numbers of women who would need to take tamoxifen to obtain the same absolute reduction in breast cancer. These findings are exploratory and need to be confirmed in other randomized trials. PMID- 12529350 TI - Retinoic acid receptor beta and telomerase catalytic subunit expression in bronchial epithelium of heavy smokers. AB - Expression of retinoic acid receptor beta (RAR beta) and telomerase catalytic subunit (human telomerase reverse transcriptase [hTERT]) was analyzed in 285 bronchial biopsy specimens from 53 heavy cigarette smokers and four former smokers by using in situ hybridization. Of the 191 biopsy specimens that were positive for RAR beta, 69% expressed hTERT, whereas only 54% of the 94 RAR beta negative biopsy specimens expressed hTERT (P =.014). Because hTERT expression in bronchial tissue has been previously associated with increased risk of lung cancer, the association between RAR beta and hTERT expression suggests that RAR beta expression may be an indicator of increased risk of lung cancer in heavy smokers. PMID- 12529351 TI - Re: Calcium intake and risk of colon cancer in women and men. PMID- 12529353 TI - False-positive results in studies of changes in fiber diffraction of hair from patients with breast cancer may not be false. PMID- 12529354 TI - Monomeric 14-3-3 protein is sufficient to modulate the activity of the Drosophila slowpoke calcium-dependent potassium channel. AB - Drosophila 14-3-3zeta (D14-3-3zeta) modulates the activity of the Slowpoke calcium-dependent potassium channel (dSlo) by interacting with the dSlo binding protein, Slob. We show here that D14-3-3zeta forms dimers in vitro. Site-directed mutations in its putative dimerization interface result in a dimerization deficient form of D14-3-3zeta. Both the wild-type and dimerization-deficient forms of D14-3-3zeta bind to Slob with similar affinity and form complexes with dSlo. When dSlo and Slob are expressed in mammalian cells, the dSlo channel activity is similarly modulated by co-expression of either the wild-type or the dimerization-deficient form of D14-3-3zeta. In addition, dSlo is still modulated by wild-type D14-3-3zeta in the presence of a 14-3-3 mutant, which does not itself bind to Slob but forms heterodimers with the wild-type 14-3-3. These data, taken together, suggest that monomeric D14-3-3zeta is capable of modulating dSlo channel activity in this regulatory complex. PMID- 12529355 TI - A single point mutation reverses the donor specificity of human blood group B synthesizing galactosyltransferase. AB - Blood group A and B antigens are carbohydrate structures that are synthesized by glycosyltransferase enzymes. The final step in B antigen synthesis is carried out by an alpha1-3 galactosyltransferase (GTB) that transfers galactose from UDP-Gal to type 1 or type 2, alphaFuc1-->2betaGal-R (H)-terminating acceptors. Similarly the A antigen is produced by an alpha1-3 N-acetylgalactosaminyltransferase that transfers N-acetylgalactosamine from UDP-GalNAc to H-acceptors. Human alpha1-3 N acetylgalactosaminyltransferase and GTB are highly homologous enzymes differing in only four of 354 amino acids (R176G, G235S, L266M, and G268A). Single crystal x-ray diffraction studies have shown that the latter two of these amino acids are responsible for the difference in donor specificity, while the other residues have roles in acceptor binding and turnover. Recently a novel cis-AB allele was discovered that produced A and B cell surface structures. It had codons corresponding to GTB with a single point mutation that replaced the conserved amino acid proline 234 with serine. Active enzyme expressed from a synthetic gene corresponding to GTB with a P234S mutation shows a dramatic and complete reversal of donor specificity. Although this enzyme contains all four "critical" amino acids associated with the production of blood group B antigen, it preferentially utilizes the blood group A donor UDP-GalNAc and shows only marginal transfer of UDP-Gal. The crystal structure of the mutant reveals the basis for the shift in donor specificity. PMID- 12529356 TI - Role of the Gla and first epidermal growth factor-like domains of factor X in the prothrombinase and tissue factor-factor VIIa complexes. AB - Factor X (FX) has high structure homology with other proteins of blood coagulation such as factor IX (FIX) and factor VII (FVII). These proteins present at their amino-terminal extremity a gamma-carboxyglutamic acid containing domain (Gla domain), followed by two epidermal growth factor-like (EGF1 and EGF2) domains, an activation peptide, and a serine protease domain. After vascular damage, the tissue factor-FVIIa (TF-FVIIa) complex activates both FX and FIX. FXa interacts stoichiometrically with tissue pathway inhibitor (TFPI), regulating TF FVIIa activity by forming the TF-FVIIa-TFPI-FXa quaternary complex. Conversely, FXa boosts coagulation by its association with its cofactor, factor Va (FVa). To investigate the contribution of the Gla and EGF1 domains of FX in these complexes, FX chimeras were produced in which FIX Gla and EGF1 domains substituted the corresponding domains of FX. The affinity of the two chimeras, FX/FIX(Gla) and FX/FIX(EGF1), for the TF-FVIIa complex was markedly reduced compared with that of wild-type-FX (wt-FX) independently of the presence of phospholipids. Furthermore, the association rate constants of preformed FX/FIX(Gla)-TFPI and FX/FIX(EGF1)-TFPI complexes with TF-FVIIa were, respectively, 10- and 5-fold slower than that of wt-FXa-TFPI complex. Finally, the apparent affinity of FVa was 2-fold higher for the chimeras than for wt-FX in the presence of phospholipids and equal in their absence. These data demonstrate that FX Gla and EGF1 domains contain residues, which interact with TF-FVIIa exosites contributing to the formation of the TF-FVIIa-FX and TF-FVIIa-TFPI-FXa complexes. On the opposite, FXa Gla and EGF1 domains are not directly involved in FVa binding. PMID- 12529357 TI - Anti-angiogenic activity of the recombinant kringle domain of urokinase and its specific entry into endothelial cells. AB - Urokinase plasminogen activator (uPA) belongs to a family of proteins that contains kringle domain and plays an important role in inflammation, tissue remodeling, angiogenesis, and tumor metastasis by pericellular plasminogen activation. Kringle domains of plasminogen have been shown to demonstrate anti angiogenic and anti-tumor activities. Here, we report our investigation of the kringle domain of uPA for anti-angiogenic activity and a possible cellular mechanism of action. The recombinant kringle domain of uPA (Asp(45)-Lys(135)) (UK1) inhibited endothelial cell proliferation stimulated by basic fibroblast growth factor, vascular endothelial growth factor (VEGF), or epidermal growth factor. It also inhibited migration of endothelial cells induced by VEGF or uPA, and in vivo angiogenesis on the chick chorioallantoic membrane. It did not block plasminogen activation by activated uPA in clot lysis and chromogenic substrate assays. Neither binding of UK1 to immobilized uPA receptor nor competitive inhibition of uPA binding were confirmed by real-time interaction analysis. However, internalization of UK1 followed by translocation from cytosol to nucleus was determined to be specific to endothelial cells. It also elicited a transient increase of Ca(2+) flux of more than 2-fold within 2 min of exposure in an endothelial cell-specific manner. These results suggest that the kringle domain of uPA exhibits anti-angiogenic activity and that its anti-angiogenic activity may occur through a different mechanism from inhibition of uPA-uPA receptor interaction or uPA proteolytic activity and may be associated with endothelial cell specific internalization not mediated by the uPA receptor. PMID- 12529359 TI - Regulation of the nitric oxide reduction operon (norRVW) in Escherichia coli. Role of NorR and sigma54 in the nitric oxide stress response. AB - Nitric oxide (NO) induces NO-detoxifying enzymes in Escherichia coli suggesting sensitive mechanisms for coordinate control of NO defense genes in response to NO stress. Exposure of E. coli to sub-micromolar NO levels under anaerobic conditions rapidly induced transcription of the NO reductase (NOR) structural genes, norV and norW, as monitored by lac gene fusions. Disruption of rpoN (sigma(54)) impaired the NO-mediated induction of norV and norW transcription and NOR expression, whereas disruption of the upstream regulatory gene, norR, completely ablated NOR induction. NOR inducibility was restored to NorR null mutants by expressing NorR in trans. Furthermore, an internal deletion of the N terminal domain of NorR activated NOR expression independent of NO exposure. Neither NorR nor sigma(54) was essential for NO-mediated induction of the NO dioxygenase (flavohemoglobin) encoded by hmp. However, elevated NOR activity inhibited NO dioxygenase induction, and, in the presence of dioxygen, NO dioxygenase inhibited norV induction by NO. The results demonstrate the role of NorR as a sigma(54)-dependent regulator of norVW expression. A role for the NorR N-terminal domain as a transducer or sensor for NO is suggested. PMID- 12529358 TI - The first crystal structure of archaeal aldolase. Unique tetrameric structure of 2-deoxy-d-ribose-5-phosphate aldolase from the hyperthermophilic archaea Aeropyrum pernix. AB - A gene encoding a 2-deoxy-d-ribose-5-phosphate aldolase (DERA) homolog was identified in the hyperthermophilic Archaea Aeropyrum pernix. The gene was overexpressed in Escherichia coli, and the produced enzyme was purified and characterized. The enzyme is an extremely thermostable DERA; its activity was not lost after incubation at 100 degrees C for 10 min. The enzyme has a molecular mass of approximately 93 kDa and consists of four subunits with an identical molecular mass of 24 kDa. This is the first report of the presence of tetrameric DERA. The three-dimensional structure of the enzyme was determined by x-ray analysis. The subunit folds into an alpha/beta-barrel. The asymmetric unit consists of two homologous subunits, and a crystallographic 2-fold axis generates the functional tetramer. The main chain coordinate of the monomer of the A. pernix enzyme is quite similar to that of the E. coli enzyme. There was no significant difference in hydrophobic interactions and the number of ion pairs between the monomeric structures of the two enzymes. However, a significant difference in the quaternary structure was observed. The area of the subunit subunit interface in the dimer of the A. pernix enzyme is much larger compared with the E. coli enzyme. In addition, the A. pernix enzyme is 10 amino acids longer than the E. coli enzyme in the N-terminal region and has an additional N terminal helix. The N-terminal helix produces a unique dimer-dimer interface. This promotes the formation of a functional tetramer of the A. pernix enzyme and strengthens the hydrophobic intersubunit interactions. These structural features are considered to be responsible for the extremely high stability of the A. pernix enzyme. This is the first description of the structure of hyperthermophilic DERA and of aldolase from the Archaea domain. PMID- 12529360 TI - Evidence for a monomeric intermediate in the reversible unfolding of F factor TraM. AB - F factor TraM is essential for efficient bacterial conjugation, but its molecular function is not clear. Because the physical properties of TraM may provide clues to its role in conjugation, we have characterized the TraM oligomerization equilibrium. We show that the reversible unfolding transition is non-two-state, indicating the presence of at least one intermediate. Analytical ultracentrifugation experiments indicate that the first phase of unfolding involves dissociation of the tetramer into folded monomers, which are subsequently unfolded to the denatured state in the second phase. Furthermore, we show that a C-terminal domain isolated by limited proteolysis is tetrameric in solution, like the full-length protein, and that its loss of structure correlates with dissociation of the TraM tetramer. Unfolding of the individual domains indicates that the N- and C-terminal regions act cooperatively to stabilize the full-length protein. Together, these experiments suggest structural overlap of regions important for oligomerization and DNA binding. We propose that modulating the oligomerization equilibrium of TraM may regulate its essential activity in bacterial conjugation. PMID- 12529361 TI - Congo red populates partially unfolded states of an amyloidogenic protein to enhance aggregation and amyloid fibril formation. AB - Congo red (CR) has been reported to inhibit or enhance amyloid fibril formation by several proteins. To gain insight into the mechanism(s) for these apparently paradoxical effects, we studied as a model amyloidogenic protein, a dimeric immunoglobulin light chain variable domain. With a range of molar ratios of CR, i.e. r = [CR]/[protein dimer], we investigated the aggregation kinetics, conformation, hydrogen-deuterium exchange, and thermal stability of the protein. In addition, we used isothermal titration calorimetry to characterize the thermodynamics of CR binding to the protein. During incubation at 37 degrees C or during thermal scanning, with CR at r = 0.3, 1.3, and 4.8, protein aggregation was greatly accelerated compared with that measured in the absence of the dye. In contrast, with CR at r = 8.8, protein unfolding was favored over aggregation. The aggregates formed with CR at r = 0 or 0.3 were typical amyloid fibrils, but mixtures of amyloid fibrils and amorphous aggregates were formed at r = 1.3 and 4.8. CR decreased the apparent thermal unfolding temperature of the protein. Furthermore, CR perturbed the tertiary structure of the protein without significantly altering its secondary structure. Consistent with this result, CR also increased the rate of hydrogen-deuterium exchange by the protein. Isothermal titration calorimetry showed that CR binding to the protein was enthalpically driven, indicating that binding was mainly the result of electrostatic interactions. Overall, these results demonstrate that at low concentrations, CR binding to the protein favors a structurally perturbed, aggregation-competent species, resulting in acceleration of fibril formation. At high CR concentration, protein unfolding is favored over aggregation, and fibril formation is inhibited. Because low concentrations of CR can promote amyloid fibril formation, the therapeutic utility of this compound or its analogs to inhibit amyloidoses is questionable. PMID- 12529362 TI - RNA interference reveals that endogenous Xenopus MinK-related peptides govern mammalian K+ channel function in oocyte expression studies. AB - The physiological properties of most ion channels are defined experimentally by functional expression of their pore-forming alpha subunits in Xenopus laevis oocytes. Here, we cloned a family of Xenopus KCNE genes that encode MinK-related peptide K(+) channel beta subunits (xMiRPs) and demonstrated their constitutive expression in oocytes. Electrophysiological analysis of xMiRP2 revealed that when overexpressed this gene modulates human cardiac K(+) channel alpha subunits HERG (human ether-a-go-go-related gene) and KCNQ1 by suppressing HERG currents and removing the voltage dependence of KCNQ1 activation. The ability of endogenous levels of xMiRP2 to contribute to the biophysical attributes of overexpressed mammalian K(+) channels in oocyte studies was assessed next. Injection of an xMiRP2 sequence-specific short interfering RNA (siRNA) oligo reduced endogenous xMiRP2 expression 5-fold, whereas a control siRNA oligo had no effect, indicating the effectiveness of the RNA interference technique in Xenopus oocytes. The functional effects of endogenous xMiRP2 silencing were tested using electrophysiological analysis of heterologously expressed HERG channels. The RNA interference-mediated reduction of endogenous xMiRP2 expression increased macroscopic HERG current as much as 10-fold depending on HERG cRNA concentration. The functional effects of human MiRP1 (hMiRP1)/HERG interaction were also affected by endogenous xMiRP2. At high HERG channel density, at which the effects of endogenous xMiRP2 are minimal, hMiRP1 reduced HERG current. At low HERG current density, hMiRP1 paradoxically up-regulated HERG current, a result consistent with hMiRP1 rescuing HERG from suppression by endogenous xMiRP2. Thus, endogenous Xenopus MiRP subunits contribute to the base-line properties of K(+) channels like HERG in oocyte expression studies, which could explain expression level- and expression system-dependent variation in K(+) channel function. PMID- 12529363 TI - Extended core 1 and core 2 branched O-glycans differentially modulate sialyl Lewis X-type L-selectin ligand activity. AB - It has been established that sialyl Lewis x in core 2 branched O-glycans serves as an E- and P-selectin ligand. Recently, it was discovered that 6-sulfosialyl Lewis x in extended core 1 O-glycans, NeuNAcalpha2-->3Galbeta1-->4(Fucalpha1- >3(sulfo-->6))GlcNAcbeta1--> 3Galbeta1-->3GalNAcalpha1-->Ser/Thr, functions as an L-selectin ligand in high endothelial venules. Extended core 1 O-glycans can be synthesized when a core 1 extension enzyme is present. In this study, we first show that beta1,3-N-acetylglucosaminyltransferase-3 (beta3GlcNAcT-3) is almost exclusively responsible for core 1 extension among seven different beta3GlcNAcTs and thus acts on core 1 O-glycans attached to PSGL-1. We found that transcripts encoding beta3GlcNAcT-3 were expressed in human neutrophils and lymphocytes but that their levels were lower than those of transcripts encoding core 2 beta1,6-N acetylglucosaminyltransferase I (Core2GlcNAcT-I). Neutrophils also expressed transcripts encoding fucosyltransferase VII (FucT-VII) and Core2GlcNAcT-I, whereas lymphocytes expressed only small amounts of transcripts encoding FucT VII. To determine the roles of sialyl Lewis x in extended core 1 O-glycans, Chinese hamster ovary (CHO) cells were stably transfected to express PSGL-1, FucT VII, and either beta3GlcNAcT-3 or Core2GlcNAcT-I. Glycan structural analyses disclosed that PSGL-1 expressed in these transfected cells carried comparable amounts of sialyl Lewis x in extended core 1 and core 2 branched O-glycans. In a rolling assay, CHO cells expressing sialyl Lewis x in extended core 1 O-glycans supported a significant degree of shear-dependent tethering and rolling of neutrophils and lymphocytes, although less than CHO cells expressing sialyl Lewis x in core 2 branched O-glycans. These results indicate that sialyl Lewis x in extended core 1 O-glycans can function as an L-selectin ligand and is potentially involved in neutrophil adhesion on neutrophils bound to activated endothelial cells. PMID- 12529364 TI - Characterization of a novel and specific inhibitor for the pro-apoptotic protease Omi/HtrA2. AB - Omi/HtrA2 is a mammalian serine protease with high homology to bacterial HtrA chaperones. Omi/HtrA2 is localized in mitochondria and is released to the cytoplasm in response to apoptotic stimuli. Omi/HtrA2 induces cell death in a caspase-dependent manner by interacting with the inhibitor of apoptosis protein as well as in a caspase-independent manner that relies on its protease activity. We describe the identification and characterization of a novel compound as a specific inhibitor of the proteolytic activity of Omi/HtrA2. This compound (ucf 101) was isolated in a high throughput screening of a combinatorial library using bacterially made Omi-(134-458) protease and fluorescein-casein as a generic substrate. ucf-101 showed specific activity against Omi/HtrA2 and very little activity against various other serine proteases. This compound has a natural fluorescence that was used to monitor its ability to enter mammalian cells. ucf 101, when tested in caspase-9 (-/-) null fibroblasts, was found to inhibit Omi/HtrA2-induced cell death. PMID- 12529365 TI - A mutation in the cystic fibrosis transmembrane conductance regulator generates a novel internalization sequence and enhances endocytic rates. AB - Cystic fibrosis is a common lethal genetic disease among Caucasians. The cystic fibrosis gene encodes a cyclic adenosine monophosphate-activated chloride channel (cystic fibrosis transmembrane conductance regulator (CFTR)) that mediates electrolyte transport across the luminal surfaces of a variety of epithelial cells. Mutations in CFTR fall into two broad categories; those that affect protein biosynthesis/stability and traffic to the cell surface and those that cause altered channel kinetics in proteins that reach the cell surface. Here we report a novel mechanism by which mutations in CFTR give rise to disease. N287Y, a mutation within an intracellular loop of CFTR, increases channel endocytosis from the cell surface without affecting either biosynthesis or channel gating. The sole consequence of this novel mutation is to generate a novel tyrosine-based endocytic sequence within an intracellular loop in CFTR leading to increased removal from the cell surface and a reduction in the steady-state level of CFTR at the cell surface. PMID- 12529366 TI - Mitochondria efficiently buffer subplasmalemmal Ca2+ elevation during agonist stimulation. AB - In endothelial cells, local Ca(2+) release from superficial endoplasmic reticulum (ER) activates BK(Ca) channels. The resulting hyperpolarization promotes capacitative Ca(2+) entry (CCE), which, unlike BK(Ca) channels, is inhibited by high Ca(2+). To understand how the coordinated activation of plasma membrane ion channels with opposite Ca(2+) sensitivity is orchestrated, the individual contribution of mitochondria and ER in regulation of subplasmalemmal Ca(2+) concentration ([Ca(2+)](pm)) was investigated. For organelle visualization, cells were transfected with DsRed and yellow cameleon targeted to mitochondria and ER. The patch pipette was placed far from any organelle (L1), close to ER (L3), or mitochondria (L2) and activity of BK(Ca) channels was used to estimate local [Ca(2+)](pm). Under standard patch conditions (130 mm K(+) in the bath), histamine increased [Ca(2+)](pm) at L1 and L3 to approximately 1.6 microm, whereas close to mitochondria [Ca(2+)](pm) remained unchanged. If mitochondria moved apart from the pipette or in the presence of carbonyl cyanide-4 trifluoromethoxyphenylhyrazone, [Ca(2+)](pm) at L2 increased in response to histamine. Under standard patch conditions Ca(2+) entry was negligible due to cell depolarization. Using a physiological patch approach (5.6 mm K(+) in the bath), changes in [Ca(2+)](pm) to histamine could be monitored without cell depolarization and, thus, in conditions where Ca(2+) entry occurred. Here, histamine induced an initial transient Ca(2+) elevation to > or =3.5 microm followed by a long lasting plateau at approximately 1.2 microm in L1 and L3, whereas mitochondria kept neighboring [Ca(2+)](pm) low during stimulation. Thus, superficial mitochondria and ER generate local domains of low and high Ca(2+) allowing simultaneous activation of BK(Ca) and CCE, despite their opposite Ca(2+) sensitivity. PMID- 12529367 TI - Role of peroxidoxins in Leishmania chagasi survival. Evidence of an enzymatic defense against nitrosative stress. AB - The mechanisms by which Leishmania parasites survive exposure to highly reactive oxygen (ROS) and nitrogen (RNS) species within phagosomes of macrophages are not well known. Recently it has been shown that RNS alone is sufficient and necessary to control Leishmania donovani infection in mice (Murray, H. W., and Nathan, C. F. (1999) J. Exp. Med. 189, 741-746). No enzymatic defense against RNS has been discovered in Leishmania to date. We have previously isolated two peroxidoxins (LcPxn1 and LcPxn2) from Leishmania chagasi and showed that recombinant LcPxn1 protein was capable of detoxifying hydrogen peroxide, hydroperoxide, and hydroxyl radicals (Barr, S. D., and Gedamu, L. (2001) J. Biol. Chem. 276, 34279-34287). In further characterizing the physiological role of peroxidoxins in Leishmania survival, we show here that recombinant LcPxn1 protein can detoxify RNS in addition to ROS, whereas recombinant LcPxn2 protein can only detoxify hydrogen peroxide. LcPxn1 and LcPxn2 are localized to the cytoplasm, and overexpression of LcPxn1 in L. chagasi parasites enhanced survival when exposed to exogenous ROS and RNS and enhanced survival within U937 macrophage cells. Site-directed mutagenesis studies revealed that the conserved Cys-52 residue is essential for detoxifying hydrogen peroxide, t-butyl hydroperoxide, and hydroxyl radicals, whereas the conserved Cys-173 residue is essential for detoxifying t-butyl hydroperoxide and peroxynitrite. This is the first report of an enzymatic defense against RNS in Leishmania. PMID- 12529368 TI - Structure and enzymatic properties of a stable complex of the human REV1 and REV7 proteins. AB - With yeast Saccharomyces cerevisiae, results from a variety of genetic and biochemical investigations have demonstrated that the REV genes play a major role in induction of mutations through replication processes that directly copy the damaged DNA template during DNA replication. However, in higher eucaryotes functions of homologues are poorly understood and appear somewhat different from the yeast case. It has been suggested that human REV1 interacts with human REV7, this being specific to higher eucaryotes. Here we show that purified human REV1 and REV7 proteins form a heterodimer in solution, which is stable through intensive purification steps. Results from biochemical analysis of the transferase reactions of the REV1-REV7 complex demonstrated, in contrast to the case of yeast Rev3 whose polymerase activity is stimulated by assembly with yeast Rev7, that human REV7 did not influence the stability, substrate specificity, or kinetic parameters of the transferase reactions of REV1 protein. The possible role of human REV7 is discussed. PMID- 12529369 TI - Interactions of SKIP/NCoA-62, TFIIB, and retinoid X receptor with vitamin D receptor helix H10 residues. AB - The vitamin D receptor (VDR) is a ligand-dependent transcription factor that heterodimerizes with retinoid X receptor (RXR) and interacts with the basal transcription machinery and transcriptional cofactors to regulate target gene activity. The p160 coactivator GRIP1 and the distinct coregulator Ski-interacting protein (SKIP)/NCoA-62 synergistically enhance ligand-dependent VDR transcriptional activity. Both coregulators bind directly to and form a ternary complex with VDR, with GRIP1 contacting the activation function-2 (AF-2) domain and SKIP/NCoA-62 interacting through an AF-2 independent interface. It was previously reported that SKIP/NCoA-62 interaction with VDR was independent of the heterodimerization interface (specifically, helices H10/H11). In contrast, the present study defines specific residues within a conserved and surface-exposed region of VDR helix H10 that are required for interaction with SKIP/NCoA-62 and for full ligand-dependent transactivation activity. SKIP/NCoA-62, the basal transcription factor TFIIB, and RXR all interacted with VDR helix H10 mutants at reduced levels compared with wild type in the absence of ligand and exhibited different degrees of increased interaction upon ligand addition. Thus, SKIP/NCoA 62 interacts with VDR at a highly conserved region not previously associated with coregulator binding to regulate transactivation by a molecular mechanism distinct from that of p160 coactivators. PMID- 12529370 TI - The metabotropic glutamate receptor mGluR5 is endocytosed by a clathrin independent pathway. AB - Metabotropic glutamate receptors 5 (mGluR5) are members of the growing group C G protein-coupled receptor family. Widely expressed in mammalian brain, they are involved in modulation of the glutamate transmission. By means of transfection of mGluR5 receptors in COS-7 cells and primary hippocampal neurons in culture followed by immunocytochemistry and quantitative image analysis and by a biochemical assay, we have studied the internalization of mGluR5 splice variants. mGluR5a and -5b were endocytosed in COS-7 cells as well as in axons and dendrites of cultured neurons. Endocytosis occurred even in the absence of receptor activity, because receptors mutated in the glutamate binding site were still internalized as well as receptors in which endogenous activity had been inhibited by an inverse agonist. We have measured a constitutive rate of endocytosis of 11.7%/min for mGluR5a. We report for the first time the endocytosis pathway of mGluR5. Internalization of mGluR5 is not mediated by clathrin-coated pits. Indeed, inhibition of this pathway by Eps15 dominant negative mutants did not disturb their endocytosis. However, the large GTPase dynamin 2 is implicated in the endocytosis of mGluR5 in COS-7. mGluR5 is the first shown member of the group C G-protein coupled receptor family internalized by a nonconventional pathway. PMID- 12529371 TI - p21-activated protein kinase 4 (PAK4) interacts with the keratinocyte growth factor receptor and participates in keratinocyte growth factor-mediated inhibition of oxidant-induced cell death. AB - Keratinocyte growth factor (KGF), a member of the fibroblast growth factor (FGF) family (also known as FGF-7), is an important protective factor for epithelial cells. The receptor for KGF (also called FGFR2-IIIb), which has intrinsic tyrosine kinase activity, is expressed specifically on epithelial cells and in the lung epithelium. Administration of KGF has been shown to protect the lung from various insults, but the mechanism of protection is not well understood. To understand the mechanism by which KGF exerts protective functions on epithelial cells, we used the yeast two-hybrid assay to identify proteins that interact with the KGF receptor (KGFR). Here we show that the cytoplasmic domain of KGFR interacts with p21-activated protein kinase (PAK) 4, which is a new member of the PAK family. The PAKs are regulated by the Rho-family GTPases Rac and Cdc42. PAK4 is the most divergent member of the PAK family of proteins and may have distinct functions. However, stimuli that regulate PAK4 activity are not known. Our data show that PAK4 can associate with the KGFR, which is dependent on KGFR tyrosine kinase activity. We show that a dominant negative mutant of PAK4 blocks KGF mediated inhibition of caspase-3 activation in epithelial cells subjected to oxidant stress. Our data demonstrate that PAK4 is an important mediator of the anti-apoptotic effects of KGF on epithelial cells. PMID- 12529372 TI - Increased Sp1-dependent transactivation of the LAMgamma 1 promoter in hepatic stellate cells co-cultured with HepG2 cells overexpressing cytochrome P450 2E1. AB - Laminin is a basement-membrane protein that increases in liver fibrosis. To study the role of oxidative stress on laminin expression, hepatic stellate cells (HSC) were co-cultured with HepG2 cells that do or do not express (E47 or C34 cells, respectively) CYP2E1, a potent generator of oxygen radicals. Co-incubation of HSC with E47 cells increased laminin beta1 and gamma1 proteins compared with co incubation with C34 cells; this increase was prevented by antioxidants and CYP2E1 inhibitors. Similar results were observed in co-culture with primary hepatocytes from saline- or pyrazole-treated (with high levels of CYP2E1) rats. Laminin alpha1 chain was not detectable in the HSC in any of the systems; however, laminin alpha2 chain increased in HSC co-cultured with E47 cells. Synthesis but not turnover of laminin beta1 and gamma1 proteins was increased in HSC in the E47 co-culture. Laminin beta1 and gamma1 mRNAs were up-regulated in HSC in the E47 co culture because of transcriptional activation of both genes. Transfection experiments in HSC with reporter constructs driven by the laminin gamma1 promoter showed maximal responsiveness with the -230/+106 and the -1400/+106 constructs in the E47 system. Gel-shift assays demonstrated an increase in Sp1 binding to the laminin gamma1 promoter in HSC when co-incubated with E47 cells, which was blocked by an anti-Sp1 antibody. Co-transfection of a Sp1 expression vector further increased the responsiveness of the -330LAMgamma1-CAT reporter vector in HSC in the HSC/E47 system. These results show that diffusable CYP2E1-derived oxidative-stress mediators induce synthesis of laminins by a transcriptional mechanism in HSC. Such interactions between hepatocytes and HSC may be important during liver fibrosis. PMID- 12529373 TI - Heterodimerization of alpha 2A- and beta 1-adrenergic receptors. AB - beta- and alpha(2)-adrenergic receptors are known to exhibit substantial cross talk and mutual regulation in tissues where they are expressed together. We have found that the beta(1)-adrenergic receptor (beta(1)AR) and alpha(2A)-adrenergic receptor (alpha(2A)AR) heterodimerize when coexpressed in cells. Immunoprecipitation studies with differentially tagged beta(1)AR and alpha(2A)AR expressed in HEK-293 cells revealed robust co-immunoprecipitation of the two receptors. Moreover, agonist stimulation of alpha(2A)AR was found to induce substantial internalization of coexpressed beta(1)AR, providing further evidence for a physical association between the two receptors in a cellular environment. Ligand binding assays examining displacement of [(3)H]dihydroalprenolol binding to the beta(1)AR by various ligands revealed that beta(1)AR pharmacological properties were significantly altered when the receptor was coexpressed with alpha(2A)AR. Finally, beta(1)AR/alpha(2A)AR heterodimerization was found to be markedly enhanced by a beta(1)AR point mutation (N15A) that blocks N-linked glycosylation of the beta(1)AR as well as by point mutations (N10A/N14A) that block N-linked glycosylation of the alpha(2A)AR. These data reveal an interaction between beta(1)AR and alpha(2A)AR that is regulated by glycosylation and that may play a key role in cross-talk and mutual regulation between these receptors. PMID- 12529374 TI - The proprotein convertase PC5A and a metalloprotease are involved in the proteolytic processing of the neural adhesion molecule L1. AB - The transmembrane and multidomain neural adhesion molecule L1 plays important functional roles in the developing and adult nervous system. L1 is proteolytically processed at two distinct sites within the extracellular domain, leading to the generation of different fragments. In this report, we present evidence that the proprotein convertase PC5A is the protease that cleaves L1 in the third fibronectin type III domain, whereas the proprotein convertases furin, PC1, PC2, PACE4, and PC7 are not effective in cleaving L1. Analysis of mutations revealed Arg(845) to be the site of cleavage generating the N-terminal 140-kDa fragment. This fragment was present in the hippocampus, which expresses PC5A, but was not detectable in the cerebellum, which does not express PC5A. The 140-kDa L1 fragment was found to be tightly associated with the full-length 200-kDa L1 molecule. The complex dissociated from the membrane upon cleavage by a protease acting at a more membrane-proximal site of full-length L1. This proteolytic cleavage was inhibited by the metalloprotease inhibitor GM 6001 and enhanced by a calmodulin inhibitor. L1-dependent neurite outgrowth of cerebellar neurons was inhibited by GM 6001, suggesting that proteolytic processing of L1 by a metalloprotease is involved in neurite outgrowth. PMID- 12529375 TI - The N-terminal end of Bax contains a mitochondrial-targeting signal. AB - The translocation of Bax alpha, a pro-apoptotic member of the BCL-2 family from the cytosol to mitochondria, is a central event of the apoptotic program. We report here that the N-terminal (NT) end of Bax alpha, which contains its first alpha helix (Eta alpha 1), is a functional mitochondrial-addressing signal both in mammals and in yeast. Similar results were obtained with a newly described variant of Bax called Bax psi, which lacks the first 20 amino acids of Bax alpha and is constitutively associated with mitochondria. Deletion of Eta alpha 1 impairs the binding of Bax psi to mitochondria, whereas a fusion of the N terminus of Bax alpha, which contains Eta alpha 1 with a cytosolic protein, results in the binding of the chimeric proteins to mitochondria both in a cell free assay and in vitro. More importantly, the mitochondria-bound chimeric proteins inhibit the interaction of Bax psi with mitochondria as well as Bax apoptogenic properties. The mutations of the Eta alpha 1, which inhibit Bax alpha and Bax psi translocation to mitochondria, also block the subsequent activation of the execution phase of apoptosis. Conversely, a deletion of the C terminus does not appear to influence Bax alpha and Bax psi mitochondrial addressing. Taken together, our results suggest that Bax is targeted to mitochondria by its NT and thus through a pathway that is unique for a member of the BCL-2 family. PMID- 12529376 TI - Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation. AB - Matrix metalloproteinases (MMPs) are essential for proper extracellular matrix remodeling, a process that takes place during obesity-mediated adipose tissue formation. Here, we examine expression profiles and the potential role of MMPs and their tissue inhibitors (TIMPs) in adipose tissue remodeling during obesity. Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity (ob/ob and db/db mice) and in a diet-induced model of obesity (AKR mice). Of the MMPs and TIMPs studied, mRNA levels for MMP-2, MMP-3, MMP-12, MMP-14, MMP-19, and TIMP-1 are strongly induced in obese adipose tissues compared with lean tissues. In contrast, MMP-7 and TIMP-3 mRNAs are markedly decreased in obesity. Interestingly, enzymatic activities of MMP-12 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions, demonstrating that MMP/TIMP balance is shifted toward increased matrix degradation in obesity. Finally, we analyze the modulation of MMP-2, MMP-19, and TIMP-1 during 3T3-L1 preadipocyte differentiation, and we explore the effect of inhibition of MMP activity on in vitro adipogenesis. We find that the synthetic MMP inhibitor BB-94 (Batimastat) decreases adipose conversion of 3T3-L1 and primary rat preadipocytes. BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of CCAAT/enhancer binding protein beta, a transcription factor that is thought to play a major role in the adipogenic program. Such findings support a role for the MMP/TIMP system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development. PMID- 12529377 TI - Uncleaved BAP31 in association with A4 protein at the endoplasmic reticulum is an inhibitor of Fas-initiated release of cytochrome c from mitochondria. AB - BAP31 is a polytopic integral protein of the endoplasmic reticulum membrane and, like BID, is a preferred substrate of caspase-8. Upon Fas/CD95 stimulation, BAP31 is cleaved within its cytosolic domain, generating proapoptotic p20 BAP31. In human KB epithelial cells expressing the caspase-resistant mutant crBAP31, Fas stimulation resulted in cleavage of BID and insertion of BAX into mitochondrial membrane, but subsequent oligomerization of BAX and BAK, egress of cytochrome c to the cytosol, and apoptosis were impaired. Bap31-null mouse cells expressing crBAP31 cannot generate the endogenous p20 BAP31 cleavage product, yet crBAP31 conferred resistance to cellular condensation and cytochrome c release in response to activation of ectopic FKBPcasp8 by FK1012z. Full-length BAP31, therefore, is a direct inhibitor of these caspase-8-initiated events, acting independently of its ability to sequester p20, with which it interacts. Employing a novel split ubiquitin yeast two-hybrid screen for BAP31-interacting membrane proteins, the putative ion channel protein of the endoplasmic reticulum, A4, was detected and identified as a constitutive binding partner of BAP31 in human cells. Ectopic A4 that was introduced into A4-deficient cells cooperated with crBAP31 to resist Fas-induced egress of cytochrome c from mitochondria and cytoplasmic apoptosis. PMID- 12529378 TI - A novel antioxidant and antiapoptotic role of omeprazole to block gastric ulcer through scavenging of hydroxyl radical. AB - The mechanism of the antiulcer effect of omeprazole was studied placing emphasis on its role to block oxidative damage and apoptosis during ulceration. Dose response studies on gastroprotection in stress and indomethacin-induced ulcer and inhibition of pylorus ligation-induced acid secretion indicate that omeprazole significantly blocks gastric lesions at lower dose (2.5 mg/kg) without inhibiting acid secretion, suggesting an independent mechanism for its antiulcer effect. Time course studies on gastroprotection and acid reduction also indicate that omeprazole almost completely blocks lesions at 1 h when acid inhibition is partial. The severity of lesions correlates well with the increased level of endogenous hydroxyl radical (*OH), which when scavenged by dimethyl sulfoxide causes around 90% reduction of the lesions, indicating that *OH plays a major role in gastric damage. Omeprazole blocks stress-induced increased generation of *OH and associated lipid peroxidation and protein oxidation, indicating that its antioxidant role plays a major part in preventing oxidative damage. Omeprazole also prevents stress-induced DNA fragmentation, suggesting its antiapoptotic role to block cell death during ulceration. The oxidative damage of DNA by *OH generated in vitro is also protected by omeprazole or its analogue, lansoprazole. Lansoprazole when incubated in a *OH-generating system scavenges *OH to produce four oxidation products of which the major one in mass spectroscopy shows a molecular ion peak at m/z 385, which is 16 mass units higher than that of lansoprazole (m/z 369). The product shows no additional aromatic proton signal for aromatic hydroxylation in (1)H NMR. The product absorbing at 278 nm shows no alkaline shift for phenols, thereby excluding the formation of hydroxylansoprazole. The product is assigned to lansoprazole sulfone formed by the addition of one oxygen atom at the sulfur center following attack by the *OH. Thus, omeprazole plays a significant role in gastroprotection by acting as a potent antioxidant and antiapoptotic molecule. PMID- 12529379 TI - Critical intracellular Ca2+ dependence of transient receptor potential melastatin 2 (TRPM2) cation channel activation. AB - TRPM2 is a member of the melastatin-related TRP (transient receptor potential) subfamily. It is expressed in brain and lymphocytes and forms a cation channel that is activated by intracellular ADP-ribose and associated with cell death. In this study we investigated the calcium dependence of human TRPM2 expressed under a tetracycline-dependent promoter in HEK-293 cells. TRPM2 expression was associated with enhanced hydrogen peroxide-evoked intracellular calcium signals. In whole-cell patch clamp recordings, switching from barium- to calcium containing extracellular solution markedly activated TRPM2 as long as ADP-ribose was in the patch pipette and exogenous intracellular calcium buffering was minimal. We suggest this effect reveals a critical dependence of TRPM2 channel activity on intracellular calcium. In the absence of extracellular calcium we observed concentration-dependent activation of TRPM2 channels by calcium delivered from the patch pipette (EC(50) 340 nM, slope 4.9); the maximum effect was at least as large as that evoked by extracellular calcium. Intracellular dialysis of cells with high concentrations of EGTA or 1,2-bis(o Aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) strongly reduced the amplitude of the extracellular calcium response, and the residual response was abolished by a mixture of high and low affinity calcium buffers. TRPM2 channel currents in inside-out patches showed a strong requirement for Ca(2+) at the intracellular face of the membrane. We suggest that calcium entering via TRPM2 proteins acts at an intracellular calcium sensor closely associated with the channel, providing essential positive feedback for channel activation. PMID- 12529380 TI - Overexpression of WISP-1 down-regulated motility and invasion of lung cancer cells through inhibition of Rac activation. AB - Wnt-induced-secreted-protein-1 (WISP-1) is a cysteine-rich, secreted factor belonging to the CCN family. These proteins have been implicated in the inhibition of metastasis; however, the mechanisms involved have not been described. We demonstrated that overexpression of WISP-1 in H460 lung cancer cells inhibited lung metastasis and in vitro cell invasion and motility. We investigated the possibility that WISP-1 may regulate activation of Rac, a small GTPase important for cytoskeletal reorganizations during motility. In an indirect assay, WISP-1-expressing cells exhibited marked reduction in Rac activation compared with control cells. Blocking antibodies to alpha(v)beta(5) and alpha(1) integrins restored Rac activation in WISP-1 cells, suggesting that the inhibitory effect of WISP-1 on Rac lies downstream of integrins. Constitutively activated Rac mutant (RacG12V) was transfected into WISP-1 cells to restore Rac activation and these WISP-1/RacG12V transfectants were used for further studies. We performed microarray and real-time PCR analyses to identify genes involved in invasion that may be differentially regulated by WISP-1. Here, we showed decreased expression of metalloproteinase-1 (MMP-1) in WISP-1 cells compared with controls but increased expression in WISP-1/RacG12V cells. In an invasion assay across collagen I, an MMP-1 target matrix, WISP-1 cells were significantly less invasive compared with controls, whereas WISP-1/RacG12V cells showed elevated invasion levels. This work illustrates a negatively regulated pathway by WISP-1 involving integrins and Rac in the down-regulation of invasion. PMID- 12529381 TI - Insertion of the membrane-proximal region of the neuronal SNARE coiled coil into the membrane. AB - In the neuron, soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins assemble into an alpha-helical coiled coil that bridges the synaptic vesicle to the plasma membrane and drives membrane fusion, a required process for neurotransmitter release at the nerve terminal. How does coiled coil formation drive membrane fusion? To investigate the structural and energetic coupling between the coiled coil and membrane, the recombinant SNARE complex in the phospholipid bilayer was studied using fluorescence quenching and site-directed spin labeling EPR. Fluorescence analysis revealed that two native Trp residues at the membrane-proximal region of the coiled coil are inserted into the membrane, tightly coupling the coiled coil to the membrane. The EPR results indicate that the coiled coil penetrates into the membrane with an oblique angle, providing a favorable geometry for the basic residues to interact with negatively charged lipids. The result supports the proposition that core complex formation directly leads to the apposition of two membranes, which could facilitate lipid mixing. Trp residues and basic residues are abundant at the membrane-proximal region of transmembrane SNARE proteins, suggesting the generality of the proposed mechanism for the SNARE complex-membrane coupling. PMID- 12529382 TI - Cytosolic phospholipase A2 is an effector of Jak/STAT signaling and is involved in platelet-derived growth factor BB-induced growth in vascular smooth muscle cells. AB - Platelet-derived growth factor-BB (PDGF-BB) is a potent mitogen and chemoattractant for vascular smooth muscle cells (VSMC). To understand its mitogenic and chemotactic signaling events, we studied the role of cytosolic phospholipase A(2) (cPLA(2)) and the Jak/STAT pathway. PDGF-BB induced the expression and activity of cPLA(2) in a time-dependent manner in VSMC. Arachidonyl trifluoromethyl ketone, a potent and specific inhibitor of cPLA(2), significantly reduced PDGF-BB-induced arachidonic acid release and DNA synthesis. PDGF-BB stimulated tyrosine phosphorylation of Jak-2 in a time-dependent manner. In addition, PDGF-BB activated STAT-3 as determined by its tyrosine phosphorylation, DNA-binding activity, and reporter gene expression, and these responses were suppressed by AG490, a selective inhibitor of Jak-2. AG490 and a dominant-negative mutant of STAT-3 also attenuated PDGF-BB-induced expression of cPLA(2,) arachidonic acid release, and DNA synthesis in VSMC. Together, these results suggest that induction of expression of cPLA(2) and arachidonic acid release are involved in VSMC growth in response to PDGF-BB and that these events are mediated by Jak-2-dependent STAT-3 activation. PMID- 12529383 TI - Dimer-specific potentiation of NGFI-B (Nur77) transcriptional activity by the protein kinase A pathway and AF-1-dependent coactivator recruitment. AB - The NGFI-B (Nur77) subfamily of orphan nuclear receptors (NRs), which also includes Nurr1 and NOR1, bind the NurRE regulatory element as either homo- or heterodimers formed between subfamily members. These NRs mediate the activation of pituitary proopiomelanocortin (POMC) gene transcription by the hypothalamic hormone corticotropin-releasing hormone (CRH), an important link between neuronal and endocrine components of the hypothalamo-pituitary-adrenal axis. CRH effects on POMC transcription do not require de novo protein synthesis. We now show that CRH signals activate Nur factors through the cyclic AMP/protein kinase A (PKA) pathway. CRH and PKA rapidly increase nuclear DNA binding activity of NGFI-B dimers but not monomers. Accordingly, CRH- or PKA-activated Nur factors enhance dimer (but not monomer) target response elements. We also show that p160/SRC coactivators are recruited to Nur dimers (but not to monomers) and that coactivator recruitment to the NurRE is enhanced in response to CRH. Moreover, PKA- and coactivator-induced potentiation of NGFI-B activity are primarily exerted through the N-terminal AF-1 domain of NGFI-B. The TIF2 (SRC-2) glutamine rich domain is required for this activity. Taken together, these results indicate that Nur factors behave as endpoint effectors of the PKA signaling pathway acting through dimers and AF-1-dependent recruitment of coactivators. PMID- 12529384 TI - Proteasome inhibition results in TRAIL sensitization of primary keratinocytes by removing the resistance-mediating block of effector caspase maturation. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) exerts potent cytotoxic activity against transformed keratinocytes, whereas primary keratinocytes are relatively resistant. In several cell types, inhibition of the proteasome sensitizes for TRAIL-induced apoptosis by interference with NF-kappaB activation. Here we describe a novel intracellular mechanism of TRAIL resistance in primary cells and how this resistance is removed by proteasome inhibitors independent of NF-kappaB in primary human keratinocytes. This sensitization was not mediated at the receptor-proximal level of TRAIL DISC formation or caspase 8 activation but further downstream. Activation of caspase 3 was critical, as it only occurred when mitochondrial apoptotic pathways were activated, as reflected by Smac/DIABLO, HtrA2, and cytochrome c release. Smac/DIABLO and HtrA2 are needed to release the X-linked inhibitor-of-apoptosis protein (XIAP)-mediated block of full caspase 3 maturation. XIAP can effectively block caspase 3 maturation and, intriguingly, is highly expressed in primary but not in transformed keratinocytes. Ectopic XIAP expression in transformed keratinocytes resulted in increased resistance to TRAIL. Our data suggest that breaking of this resistance via proteasome inhibitors, which are potential anticancer drugs, may sensitize certain primary cells to TRAIL-induced apoptosis and could thereby complicate the clinical applicability of a combination of TRAIL receptor agonists with proteasome inhibitors. PMID- 12529386 TI - The dihydrofolate reductase origin of replication does not contain any nonredundant genetic elements required for origin activity. AB - The Chinese hamster dihydrofolate reductase (DHFR) origin of replication consists of a broad zone of potential initiation sites scattered throughout a 55-kb intergenic spacer, with at least three sites being preferred (ori-beta, ori beta', and ori-gamma). We previously showed that deletion of the most active site or region (ori-beta) has no demonstrable effect on initiation in the remainder of the intergenic spacer nor on the time of replication of the DHFR locus as a whole. In the present study, we have now deleted ori-beta', both ori-beta and ori beta', an 11-kb region just downstream from the DHFR gene, or the central approximately 40-kb core of the spacer. The latter two deletions together encompass >95% of the initiation sites that are normally used in this locus. Two dimensional gel analysis shows that initiation still occurs in the early S phase in the remainder of the intergenic spacer in each of these deletion variants. Even removal of the 40-kb core fails to elicit a significant effect on the time of replication of the DHFR locus in the S period; indeed, in the truncated spacer that remains, the efficiency of initiation actually appears to increase relative to the corresponding region in the wild-type locus. Thus, if replicators control the positions of nascent strand start sites in this complex origin, either (i) there must be a very large number of redundant elements in the spacer, each of which regulates initiation only in its immediate environment, or (ii) they must lie outside the central core in which the vast majority of nascent strand starts occur. PMID- 12529385 TI - Critical role for mouse Hus1 in an S-phase DNA damage cell cycle checkpoint. AB - Mouse Hus1 encodes an evolutionarily conserved DNA damage response protein. In this study we examined how targeted deletion of Hus1 affects cell cycle checkpoint responses to genotoxic stress. Unlike hus1(-) fission yeast (Schizosaccharomyces pombe) cells, which are defective for the G(2)/M DNA damage checkpoint, Hus1-null mouse cells did not inappropriately enter mitosis following genotoxin treatment. However, Hus1-deficient cells displayed a striking S-phase DNA damage checkpoint defect. Whereas wild-type cells transiently repressed DNA replication in response to benzo(a)pyrene dihydrodiol epoxide (BPDE), a genotoxin that causes bulky DNA adducts, Hus1-null cells maintained relatively high levels of DNA synthesis following treatment with this agent. However, when treated with DNA strand break-inducing agents such as ionizing radiation (IR), Hus1-deficient cells showed intact S-phase checkpoint responses. Conversely, checkpoint-mediated inhibition of DNA synthesis in response to BPDE did not require NBS1, a component of the IR-responsive S-phase checkpoint pathway. Taken together, these results demonstrate that Hus1 is required specifically for one of two separable mammalian checkpoint pathways that respond to distinct forms of genome damage during S phase. PMID- 12529388 TI - Granulocyte function in grancalcin-deficient mice. AB - Grancalcin, one of the penta-EF-hand Ca(2+) binding proteins, is expressed at high levels in polymorphonuclear granulocytes (neutrophils). EF-hand proteins are implicated in the regulation of diverse processes including cell migration, apoptosis, and mobilization of neutrophil effector functions. To determine the role of grancalcin in vivo, we inactivated the gene encoding grancalcin (Gca) in embryonic stem cells and generated grancalcin-deficient mice. Homozygous Gca mutants appeared healthy and reproduced normally. Leukocyte recruitment into the peritoneal cavity upon induction of inflammation was not significantly affected by the absence of grancalcin. The mutants also resisted systemic fungal infection similarly to wild-type mice, and in vitro killing of Staphylococcus aureus by inflammatory cells was not significantly impaired. While marginally increased survival rates of mutants faced with endotoxic shock may indicate a contribution of grancalcin to immunopathogenesis, it is not essential for vital leukocyte effector functions required to control microbial infections. PMID- 12529387 TI - Recruitment of a repressosome complex at the growth hormone receptor promoter and its potential role in diabetic nephropathy. AB - The growth hormone (GH)-GH receptor (GHR) axis modulates growth and metabolism and contributes to complications of diabetes mellitus. We analyzed the promoter region of the dominant transcript (L2) of the murine GHR to determine that a cis element, L2C1, interacts with transcription factors NF-Y, BTEB1, and HMG-Y/I. These proteins individually repress GHR expression and together form a repressosome complex in conjunction with mSin3b. The histone deacetylase inhibitor trichostatin A increases expression of the murine GHR gene, enhances association of acetyl-H3 at L2C1, inhibits formation of the repressosome complex, and decreases NF-Y's association with L2C1. Our studies reveal that murine models of experimental diabetes mellitus are characterized by reduced hepatic GHR expression, decreased acetyl-H3 associated with L2C1, and increased formation of the repressosome complex. In contrast, in the kidney diabetes mellitus is associated with enhanced GHR expression and lack of alteration in the assembly of the repressosome complex, thus permitting exposure of kidneys to the effects of elevated levels of GH in diabetes mellitus. Our findings define a higher-order repressosome complex whose formation correlates with the acetylation status of chromatin histone proteins. The delineation of the role of this repressosome complex in regulating tissue-specific expression of GHR in diabetes mellitus provides a molecular model for the role of GH in the genesis of certain microvascular complications of diabetes mellitus. PMID- 12529389 TI - Identification of homeodomain proteins, PBX1 and PREP1, involved in the transcription of murine leukemia virus. AB - Cyclin-dependent kinase inhibitors (CDKIs) have been shown to block human immunodeficiency virus and herpes simplex virus. It is hypothesized that CDKIs block viral replication by inhibiting transcription of specific cellular genes. Here we find that three CDKIs, flavopiridol, purvalanol A, and methoxy roscovitine, block Moloney murine leukemia virus (MLV) transcription events. Using gene expression microarray technology to examine the inhibitory effects of CDKIs, we observed a cellular gene, the pre-B-cell leukemia transcription factor 1 (Pbx1) gene, down-regulated by CDKI treatment. The PBX consensus element (PCE), TGATTGAC, is conserved in the long terminal repeats of several murine retroviruses, including Moloney MLV. Mutations in the PCE completely inhibited viral transcription whereas overexpression of PBX1 and a PBX1-associated protein, PREP1, enhanced viral transcription. The interaction between the PCE and PBX1 PREP1 proteins was confirmed by gel shift experiments. Blocking PBX1 protein synthesis resulted in a significant decrease in viral transcription. Collectively, our results represent the first work demonstrating that the homeodomain proteins PBX1 and PREP1 are cellular factors involved in Moloney MLV transcription regulation. PMID- 12529390 TI - Nonsense-containing mRNAs that accumulate in the absence of a functional nonsense mediated mRNA decay pathway are destabilized rapidly upon its restitution. AB - Nonsense-mediated mRNA decay (NMD) is a conserved proofreading mechanism that protects eukaryotic cells from the potentially deleterious effects of truncated proteins. Studies of Saccharomyces cerevisiae imply that NMD is a predominantly cytoplasmic decay pathway, while studies of mammalian systems suggest that decay of most substrate mRNAs may occur while they are still associated with the nucleus, possibly during a round of translation that occurs during their export to the cytoplasm. Complete entry of the latter mRNAs into the cytoplasm appears to render them immune to further NMD; i.e., they escape further susceptibility to this decay pathway. To determine if yeast cytoplasmic nonsense-containing mRNAs that evade decay are subsequently immune to NMD, we examined the consequences of placing each of the three UPF/NMD genes under the control of a galactose inducible promoter. The decay kinetics of ADE2 and PGK1 nonsense-containing mRNAs were then analyzed when expression of UPF1, NMD2, or UPF3 was either repressed or subsequently induced. Results from these experiments demonstrated that activation of NMD caused rapid and immediate degradation of both substrate transcripts, with half-lives of both stable mRNA populations shortened to approximately 7 min. These findings make it unlikely that yeast nonsense-containing mRNAs can escape degradation by NMD and indicate that such mRNAs are available to this decay pathway at each round of translation. PMID- 12529391 TI - Protein kinase C phosphorylates ribosomal protein S6 kinase betaII and regulates its subcellular localization. AB - The ribosomal protein S6 kinase (S6K) belongs to the AGC family of Ser/Thr kinases and is known to be involved in the regulation of protein synthesis and the G(1)/S transition of the cell cycle. There are two forms of S6K, termed S6Kalpha and S6Kbeta, which have cytoplasmic and nuclear splice variants. Nucleocytoplasmic shuttling has been recently proposed for S6Kalpha, based on the use of the nuclear export inhibitor, leptomycin B. However, the molecular mechanisms regulating subcellular localization of S6Ks in response to mitogenic stimuli remain to be elucidated. Here we present data on the in vitro and in vivo phosphorylation of S6Kbeta, but not S6Kalpha, by protein kinase C (PKC). The site of phosphorylation was identified as S486, which is located within the C-terminal nuclear localization signal. Mutational analysis and the use of phosphospecific antibodies provided evidence that PKC-mediated phosphorylation at S486 does not affect S6K activity but eliminates the function of its nuclear localization signal and causes retention of an activated form of the kinase in the cytoplasm. Taken together, this study uncovers a novel mechanism for the regulation of nucleocytoplasmic shuttling of S6KbetaII by PKC-mediated phosphorylation. PMID- 12529392 TI - Identification of farnesoid X receptor beta as a novel mammalian nuclear receptor sensing lanosterol. AB - Nuclear receptors are ligand-modulated transcription factors. On the basis of the completed human genome sequence, this family was thought to contain 48 functional members. However, by mining human and mouse genomic sequences, we identified FXRbeta as a novel family member. It is a functional receptor in mice, rats, rabbits, and dogs but constitutes a pseudogene in humans and primates. Murine FXRbeta is widely coexpressed with FXR in embryonic and adult tissues. It heterodimerizes with RXRalpha and stimulates transcription through specific DNA response elements upon addition of 9-cis-retinoic acid. Finally, we identified lanosterol as a candidate endogenous ligand that induces coactivator recruitment and transcriptional activation by mFXRbeta. Lanosterol is an intermediate of cholesterol biosynthesis, which suggests a direct role in the control of cholesterol biosynthesis in nonprimates. The identification of FXRbeta as a novel functional receptor in nonprimate animals sheds new light on the species differences in cholesterol metabolism and has strong implications for the interpretation of genetic and pharmacological studies of FXR-directed physiologies and drug discovery programs. PMID- 12529393 TI - Systematic mutagenesis of the Saccharomyces cerevisiae MLH1 gene reveals distinct roles for Mlh1p in meiotic crossing over and in vegetative and meiotic mismatch repair. AB - In eukaryotic cells, DNA mismatch repair is initiated by a conserved family of MutS (Msh) and MutL (Mlh) homolog proteins. Mlh1 is unique among Mlh proteins because it is required in mismatch repair and for wild-type levels of crossing over during meiosis. In this study, 60 new alleles of MLH1 were examined for defects in vegetative and meiotic mismatch repair as well as in meiotic crossing over. Four alleles predicted to disrupt the Mlh1p ATPase activity conferred defects in all functions assayed. Three mutations, mlh1-2, -29, and -31, caused defects in mismatch repair during vegetative growth but allowed nearly wild-type levels of meiotic crossing over and spore viability. Surprisingly, these mutants did not accumulate high levels of postmeiotic segregation at the ARG4 recombination hotspot. In biochemical assays, Pms1p failed to copurify with mlh1 2, and two-hybrid studies indicated that this allele did not interact with Pms1p and Mlh3p but maintained wild-type interactions with Exo1p and Sgs1p. mlh1-29 and mlh1-31 did not alter the ability of Mlh1p-Pms1p to form a ternary complex with a mismatch substrate and Msh2p-Msh6p, suggesting that the region mutated in these alleles could be responsible for signaling events that take place after ternary complex formation. These results indicate that mismatches formed during genetic recombination are processed differently than during replication and that, compared to mismatch repair functions, the meiotic crossing-over role of MLH1 appears to be more resistant to mutagenesis, perhaps indicating a structural role for Mlh1p during crossing over. PMID- 12529394 TI - Nuclear factor 1 is required for both hormone-dependent chromatin remodeling and transcriptional activation of the mouse mammary tumor virus promoter. AB - The mouse mammary tumor virus (MMTV) promoter has been used as a model to study how the glucocorticoid receptor (GR) remodels chromatin to allow other transcription factors to bind and activate transcription. To dissect the precise role of nuclear factor 1 (NF1) in chromatin remodeling and transcriptional activation, we used linker-scanning mutants of transcription factor binding sites on the MMTV promoter. We compared the NF1 mutant MMTV promoter in the context of transiently transfected templates (transient transfection) and templates organized as chromatin (stable transfection) to understand the effect of chromatin on factor binding and transcription. We show that on a transiently transfected template, mutation in the NF1 binding site reduces both basal and hormone-dependent transcription. This suggests that NF1 is required for transcription in the absence of organized chromatin. We also found that binding of NF1 on a transiently transfected template is independent of mutation in hormone response elements or the octamer transcription factor (OTF) binding site. In contrast, the binding of OTF proteins to a transiently transfected template was found to be dependent on the binding of NF1, which may imply that NF1 has a stabilizing effect on OTF binding. On a chromatin template, mutation in the NF1 binding site does not affect the positioning of nucleosomes on the promoter. We also show that in the absence of NF1 binding, GR-mediated chromatin remodeling of nucleosome B is reduced and hormone-dependent activation of transcription is abolished. Further, we demonstrate that NF1 is required for both the association of BRG1 chromatin remodeling complex and the GR on the promoter in vivo. These results suggest the novel possibility that NF1 may participate in chromatin remodeling activities in addition to directly enhancing transcription and that in the absence of its binding site the GR is unable to effectively bind the promoter and recruit the remodeling complex. PMID- 12529395 TI - In vivo association of the stability control protein alphaCP with actively translating mRNAs. AB - Posttranscriptional controls play a major role in eucaryotic gene expression. These controls are mediated by sequence-specific interactions of cis-acting determinants in target mRNAs with one or more protein factors. The positioning of a subset of these mRNA-protein (RNP) complexes within the 3' untranslated region (3' UTR) may allow them to remain associated with the mRNA during active translation. Robust expression of human alpha-globin mRNA during erythroid differentiation has been linked to formation of a binary complex between a KH domain protein, alphaCP, and a 3' UTR C-rich motif. Detection of this "alpha complex" has been limited to in vitro studies, and the functional state of the alpha-globin mRNA targeted by alphaCP has not been defined. In the present study we demonstrate that a significant fraction of alphaCP is associated with polysomal mRNA. Targeted analysis of the polysomal RNP complexes revealed that alphaCP is specifically bound to actively translating alpha-globin mRNA. The bound alphaCP is restricted to the poly(C)-rich 3' UTR motif and is dislodged when ribosomes are allowed to enter this region. These data validate the general importance of the 3' UTR as a sheltered site for RNP complexes and support a specific model in which the stabilizing function of alphaCP is mediated on actively translating target mRNAs. PMID- 12529396 TI - Disruption of the regulatory beta subunit of protein kinase CK2 in mice leads to a cell-autonomous defect and early embryonic lethality. AB - Protein kinase CK2 is a ubiquitous protein kinase implicated in proliferation and cell survival. Its regulatory beta subunit, CK2beta, which is encoded by a single gene in mammals, has been suspected of regulating other protein kinases. In this work, we show that knockout of the CK2beta gene in mice leads to postimplantation lethality. Mutant embryos were reduced in size at embryonic day 6.5 (E6.5). They did not exhibit signs of apoptosis but did show reduced cell proliferation. Mutant embryos were resorbed at E7.5. In vitro, CK2beta(-/-) morula development stopped after the blastocyst stage. Attempts to generate homozygous embryonic stem (ES) cells failed. By using a conditional knockout approach, we show that lack of CK2beta is deleterious for mouse ES cells and primary embryonic fibroblasts. This is in contrast to what occurs with yeast cells, which can survive without functional CK2beta. Thus, our study demonstrates that in mammals, CK2beta is essential for viability at the cellular level, possibly because it acquired new functions during evolution. PMID- 12529397 TI - The absence of mitochondrial thioredoxin 2 causes massive apoptosis, exencephaly, and early embryonic lethality in homozygous mice. AB - Thioredoxin 2 (Trx-2) is a small redox protein containing the thioredoxin active site Trp-Cys-Gly-Pro-Cys that is localized to the mitochondria by a mitochondrial leader sequence and encoded by a nuclear gene (Trx-2). Trx-2 plays an important role in cell viability and the regulation of apoptosis in vitro. To investigate the role of Trx-2 in mouse development, we studied the phenotype of mice that have the Trx-2 gene silenced by mutational insertion. Homozygous mutant embryos do not survive to birth and die after implantation at Theiler stage 15/16. The homozygous mutant embryos display an open anterior neural tube and show massively increased apoptosis at 10.5 days postcoitus and are not present by 12.5 days postcoitus. The timing of the embryonic lethality coincides with the maturation of the mitochondria, since they begin oxidative phosphorylation during this stage of embryogenesis. In addition, embryonic fibroblasts cultured from homozygous Trx 2-null embryos were not viable. Heterozygous mice are fertile and have no discernible phenotype visible by external observation, despite having decreased Trx-2 mRNA and protein. These results show that the mitochondrial redox protein Trx-2 is required for normal development of the mouse embryo and for actively respiring cells. PMID- 12529398 TI - Liver-specific reactivation of the inactivated Hnf-1alpha gene: elimination of liver dysfunction to establish a mouse MODY3 model. AB - Mice deficient in hepatocyte nuclear factor 1 alpha (HNF-1alpha) develop dwarfism, liver dysfunction, and type 2 diabetes mellitus. Liver dysfunction in HNF-1alpha-null mice includes severe hepatic glycogen accumulation and dyslipidemia. The liver dysfunction may appear as soon as 2 weeks after birth. Since the HNF-1alpha-null mice become diabetic 2 weeks after birth, the early onset of the liver dysfunction is unlikely to be due to the diabetic status of the mice. More likely, it is due directly to the deficiency of HNF-1alpha in liver. Although the HNF-1alpha-null mice have an average life span of 1 year, the severe liver phenotype has thwarted attempts to study the pathogenesis of maturity-onset diabetes of the young type 3 (MODY3) and to examine therapeutic strategies for diabetes prevention and treatment in these mice. To circumvent this problem, we have generated a new Hnf-1alpha mutant mouse line, Hnf 1alpha(kin/kin), using gene targeting to inactivate the Hnf-1alpha gene and at the same time, to incorporate the Cre-loxP DNA recombination system into the locus for later revival of the Hnf-1alpha gene in tissues by tissue-specifically expressed Cre recombinase. The Hnf-1alpha(kin/kin) mice in which the expression of HNF-1alpha was inactivated in germ line cells were indistinguishable from the HNF-1alpha-null mice with regard to both the diabetes and liver phenotypes. Intriguingly, when the inactivated Hnf-1alpha gene was revived in liver (hepatic Hnf-1alpha revived) by the Cre recombinase driven by an albumin promoter, the Hnf 1alpha(kin/kin) mice, although severely diabetic, grew normally and did not develop any of the liver dysfunctions. In addition, we showed that the expression of numerous genes in pancreas, including a marker gene for pancreas injury, was affected by liver dysfunction but not by the deficiency of HNF-1alpha in pancreas. Thus, our hepatic-Hnf-1alpha-revived mice may serve as a useful mouse model to study the human MODY3 disorder. PMID- 12529399 TI - R-Ras promotes focal adhesion formation through focal adhesion kinase and p130(Cas) by a novel mechanism that differs from integrins. AB - R-Ras regulates integrin function, but its effects on integrin signaling pathways have not been well described. We demonstrate that activation of R-Ras promoted focal adhesion formation and altered localization of the alpha2beta1 integrin from cell-cell to cell-matrix adhesions in breast epithelial cells. Constitutively activated R-Ras(38V) dramatically enhanced focal adhesion kinase (FAK) and p130(Cas) phosphorylation upon collagen stimulation or clustering of the alpha2beta1 integrin, even in the absence of increased ligand binding. Signaling events downstream of R-Ras differed from integrins and K-Ras, since pharmacological inhibition of Src or disruption of actin inhibited integrin mediated FAK and p130(Cas) phosphorylation, focal adhesion formation, and migration in control and K-Ras(12V)-expressing cells but had minimal effect in cells expressing R-Ras(38V). Therefore, signaling from R-Ras to FAK and p130(Cas) has a component that is Src independent and not through classic integrin signaling pathways and a component that is Src dependent. R-Ras effector domain mutants and pharmacological inhibition suggest a partial role for phosphatidylinositol 3-kinase (PI3K), but not Raf, in R-Ras signaling to FAK and p130(Cas). However, PI3K cannot account for the Src-independent pathway, since simultaneous inhibition of both PI3K and Src did not completely block effects of R-Ras on FAK phosphorylation. Our results suggest that R-Ras promotes focal adhesion formation by signaling to FAK and p130(Cas) through a novel mechanism that differs from but synergizes with the alpha2beta1 integrin. PMID- 12529400 TI - Homeodomain-interacting protein kinase 1 modulates Daxx localization, phosphorylation, and transcriptional activity. AB - We describe an interaction between homeodomain-interacting protein kinase 1 (HIPK1) and Daxx, two transcriptional regulators important in transducing growth regulatory signals. We demonstrate that HIPK1 is ubiquitously expressed in mice and humans and localizes predominantly to the nucleus. Daxx normally resides within the nucleus in promyelocytic leukemia protein (PML) oncogenic domains (PODs), where it physically interacts with PML. Under certain circumstances, Daxx is relocalized from PODs to chromatin, where it then acts as a transcriptional repressor through an association with histone deacetylase (HDAC1). We propose two novel mechanisms for regulating the activity of Daxx, both mediated by HIPK1. First, HIPK1 physically interacts with Daxx in cells and consequently relocalizes Daxx from PODs. Daxx relocalization disrupts its interaction with PML and augments its interaction with HDAC1, likely influencing Daxx activity. Although the relocalization of Daxx from PODs is phosphorylation independent, an active HIPK1 kinase domain is required, suggesting that HIPK1 autophosphorylation is important in this interaction. Second, HIPK1 phosphorylates Daxx on Ser 669, and phosphorylation of this site is important in modulating the ability of Daxx to function as a transcriptional repressor. Mutation of Daxx Ser 669 to Ala results in increased repression in three of four transcriptional reporters, suggesting that phosphorylation by HIPK1 diminishes Daxx transcriptional repression of specific promoters. Taken together, our results indicate that HIPK1 and Daxx collaborate in regulating transcription. PMID- 12529402 TI - Live-cell fluorescence imaging reveals the dynamics of protein kinase CK2 individual subunits. AB - Protein kinase CK2 is a multifunctional enzyme which has long been described as a stable heterotetrameric complex resulting from the association of two catalytic (alpha or alpha') and two regulatory (beta) subunits. To track the spatiotemporal dynamics of CK2 in living cells, we fused its catalytic alpha and regulatory beta subunits with green fluorescent protein (GFP). Both CK2 subunits contain nuclear localization domains that target them independently to the nucleus. Imaging of stable cell lines expressing low levels of GFP-CK2alpha or GFP-CK2beta revealed the existence of CK2 subunit subpopulations exhibiting differential dynamics. Once in the nucleus, they diffuse randomly at different rates. Unlike CK2beta, CK2alpha can shuttle, showing the dynamic nature of the nucleocytoplasmic trafficking of the kinase. When microinjected in the cytoplasm, the isolated CK2 subunits are rapidly translocated into the nucleus, whereas the holoenzyme complex remains in this cell compartment, suggesting an intramolecular masking of the nuclear localization sequences that suppresses nuclear accumulation. However, binding of FGF-2 to the holoenzyme triggers its nuclear translocation. Since the substrate specificity of CK2alpha is dramatically changed by its association with CK2beta, the control of the nucleocytoplasmic distribution of each subunit may represent a unique potential regulatory mechanism for CK2 activity. PMID- 12529401 TI - The exocytotic trafficking of TC10 occurs through both classical and nonclassical secretory transport pathways in 3T3L1 adipocytes. AB - To examine the structural determinants necessary for TC10 trafficking, localization, and function in adipocytes, we generated a series of point mutations in the carboxyl-terminal targeting domain of TC10. Wild-type TC10 (TC10/WT) localized to secretory membrane compartments and caveolin-positive lipid raft microdomains at the plasma membrane. Expression of a TC10/C206S point mutant resulted in a trafficking and localization pattern that was indistinguishable from that of TC10/WT. In contrast, although TC10/C209S or the double TC10/C206,209S mutant was plasma membrane localized, it was excluded from both the secretory membrane system and the lipid raft compartments. Surprisingly, inhibition of Golgi membrane transport with brefeldin A did not prevent plasma membrane localization of TC10 or H-Ras. Moreover, inhibition of trans-Golgi network exit with a 19 degrees C temperature block did not prevent the trafficking of TC10 or H-Ras to the plasma membrane. These data demonstrate that TC10 and H-Ras can both traffic to the plasma membrane by at least two distinct transport mechanisms in adipocytes, one dependent upon intracellular membrane transport and another independent of the classical secretory membrane system. Moreover, the transport through the secretory pathway is necessary for the localization of TC10 to lipid raft microdomains at the plasma membrane. PMID- 12529403 TI - Transcriptional activities of the zinc finger protein Zac are differentially controlled by DNA binding. AB - Zac encodes a zinc finger protein that promotes apoptosis and cell cycle arrest and is maternally imprinted. Here, we show that Zac contains transactivation and repressor activities and that these transcriptional activities are differentially controlled by DNA binding. Zac transactivation mapped to two distinct domains. One of these contained multiple repeats of the peptide PLE, which behaved as an autonomous activation unit. More importantly, we identified two related high affinity DNA-binding sites which were differentially bound by seven Zac C(2)H(2) zinc fingers. Zac bound as a monomer through zinc fingers 6 and 7 to the palindromic DNA element to confer transactivation. In contrast, binding as a monomer to one half-site of the repeat element turned Zac into a repressor. Conversely, Zac dimerization at properly spaced direct and reverse repeat elements enabled transactivation, which strictly correlated with DNA-dependent and -independent contacts of key residues within the recognition helix of zinc finger 7. The later ones support specific functional connections between Zac DNA binding and transcriptional-regulatory surfaces. Both classes of DNA elements were identified in a new Zac target gene and confirmed that the zinc fingers communicate with the transactivation function. Together, our data demonstrate a role for Zac as a transcription factor in addition to its role as coactivator for nuclear receptors and p53. PMID- 12529404 TI - Transcriptional coactivation of bone-specific transcription factor Cbfa1 by TAZ. AB - Core-binding factor 1 (Cbfa1; also called Runx2) is a transcription factor belonging to the Runt family of transcription factors that binds to an osteoblast specific cis-acting element (OSE2) activating the expression of osteocalcin, an osteoblast-specific gene. Using the yeast two-hybrid system, we identified a transcriptional coactivator, TAZ (transcriptional coactivator with PDZ-binding motif), that binds to Cbfa1. A functional relationship between Cbfa1 and TAZ is demonstrated by the coimmunoprecipitation of TAZ by Cbfa1 and by the fact that TAZ induces a dose-dependent increase in the activity of osteocalcin promoter luciferase constructs by Cbfa1. A dominant-negative construct of TAZ in which the coactivation domains have been deleted reduces osteocalcin gene expression down to basal levels. NIH 3T3, MC 3T3, and ROS 17/2.8 cells showed the expected nuclear localization of Cbfa1, whereas TAZ was distributed throughout the cytoplasm with some nuclear localization when transfected with either Cbfa1 or TAZ. Upon cotransfection by both Cbfa1 and TAZ, the transfected TAZ shows predominant nuclear localization. The dominant-negative construct of TAZ shows minimal nuclear localization upon cotransfection with Cbfa1. These data indicate that TAZ is a transcription coactivator for Cbfa1 and may be involved in the regulation of osteoblast differentiation. PMID- 12529405 TI - The carboxy terminus of Prospero regulates its subcellular localization. AB - Subcellular localization of the transcription factor Prospero is dynamic. For example, the protein is cytoplasmic in neuroblasts, nuclear in sheath cells, and degraded in newly formed neurons. The carboxy terminus of Prospero, including the homeodomain and Prospero domain, plays roles in regulating these changes. The homeodomain has two distinct subdomains, which exclude proteins from the nucleus, while the intact homeo/Prospero domain masks this effect. One subdomain is an Exportin-dependent nuclear export signal requiring three conserved hydrophobic residues, which models onto helix 1. Another, including helices 2 and 3, requires proteasome activity to degrade nuclear protein. Finally, the Prospero domain is missing in pros(I13) embryos, thus unmasking nuclear exclusion, resulting in constitutively cytoplasmic protein. Multiple processes direct Prospero regulation of cell fate in embryonic nervous system development. PMID- 12529406 TI - Interaction of histone acetylases and deacetylases in vivo. AB - Having opposing enzymatic activities, histone acetylases (HATs) and deacetylases affect chromatin and regulate transcription. The activities of the two enzymes are thought to be balanced in the cell by an unknown mechanism that may involve their direct interaction. Using fluorescence resonance energy transfer analysis, we demonstrated that the acetylase PCAF and histone deacetylase 1 (HDAC1) are in close spatial proximity in living cells, compatible with their physical interaction. In agreement, coimmunoprecipitation assays demonstrated that endogenous HDACs are associated with PCAF and another acetylase, GCN5, in HeLa cells. We found by glycerol gradient sedimentation analysis that HATs are integrated into a large multiprotein HDAC complex that is distinct from the previously described HDAC complexes containing mSin3A, Mi-2/NRD, or CoREST. This HDAC-HAT association is partly accounted for by a direct protein-protein interaction observed in vitro. The HDAC-HAT complex may play a role in establishing a dynamic equilibrium of the two enzymes in vivo. PMID- 12529407 TI - Loss of S100A9 (MRP14) results in reduced interleukin-8-induced CD11b surface expression, a polarized microfilament system, and diminished responsiveness to chemoattractants in vitro. AB - The S100A9 (MRP14) protein is abundantly expressed in myeloid cells and has been associated with various inflammatory diseases. The S100A9-deficient mice described here were viable, fertile, and generally of healthy appearance. The myelopoietic potential of the S100A9-null bone marrow was normal. S100A8, the heterodimerization partner of S100A9 was not detectable in peripheral blood cells, suggesting that even a deficiency in both S100A8 and S100A9 proteins was compatible with viable and mature neutrophils. Surprisingly, the invasion of S100A9-deficient leukocytes into the peritoneum and into the skin in vivo was indistinguishable from that in wild-type mice. However, stimulation of S100A9 deficient neutrophils with interleukin-8 in vitro failed to provoke an up regulation of CD11b. Migration upon a chemotactic stimulus through an endothelial monolayer was markedly diminished in S100A9-deficient neutrophils. Attenuated chemokinesis of the S100A9-deficient neutrophils was observed by using a three dimensional collagen matrix migration assay. The altered migratory behavior was associated with a microfilament system that was highly polarized in unstimulated S100A9-deficient neutrophils. Our data suggest that loss of the calcium-binding S100A9 protein reduces the responsiveness of the neutrophils upon chemoattractant stimuli at least in vitro. Alternative pathways for neutrophil emigration may be responsible for the lack of any effect in the two in vivo models we have investigated so far. PMID- 12529408 TI - Conditional mutation of Rb causes cell cycle defects without apoptosis in the central nervous system. AB - Targeted disruption of the retinoblastoma gene in mice leads to embryonic lethality in midgestation accompanied by defective erythropoiesis. Rb(-/-) embryos also exhibit inappropriate cell cycle activity and apoptosis in the central nervous system (CNS), peripheral nervous system (PNS), and ocular lens. Loss of p53 can prevent the apoptosis in the CNS and lens; however, the specific signals leading to p53 activation have not been determined. Here we test the hypothesis that hypoxia caused by defective erythropoiesis in Rb-null embryos contributes to p53-dependent apoptosis. We show evidence of hypoxia in CNS tissue from Rb(-/-) embryos. The Cre-loxP system was then used to generate embryos in which Rb was deleted in the CNS, PNS and lens, in the presence of normal erythropoiesis. In contrast to the massive CNS apoptosis in Rb-null embryos at embryonic day 13.5 (E13.5), conditional mutants did not have elevated apoptosis in this tissue. There was still significant apoptosis in the PNS and lens, however. Rb(-/-) cells in the CNS, PNS, and lens underwent inappropriate S-phase entry in the conditional mutants at E13.5. By E18.5, conditional mutants had increased brain size and weight as well as defects in skeletal muscle development. These data support a model in which hypoxia is a necessary cofactor in the death of CNS neurons in the developing Rb mutant embryo. PMID- 12529409 TI - Targeted disruption of the mouse Mel(1b) melatonin receptor. AB - Two high-affinity, G protein-coupled melatonin receptor subtypes have been identified in mammals. Targeted disruption of the Mel(1a) melatonin receptor prevents some, but not all, responses to the hormone, suggesting functional redundancy among receptor subtypes (Liu et al., Neuron 19:91-102, 1997). In the present work, the mouse Mel(1b) melatonin receptor cDNA was isolated and characterized, and the gene has been disrupted. The cDNA encodes a receptor with high affinity for melatonin and a pharmacological profile consistent with its assignment as encoding a melatonin receptor. Mice with targeted disruption of the Mel(1b) receptor have no obvious circadian phenotype. Melatonin suppressed multiunit electrical activity in the suprachiasmatic nucleus (SCN) in Mel(1b) receptor-deficient mice as effectively as in wild-type controls. The neuropeptide, pituitary adenylyl cyclase activating peptide, increases the level of phosphorylated cyclic AMP response element binding protein (CREB) in SCN slices, and melatonin reduces this effect. The Mel(1a) receptor subtype mediates this inhibitory response at moderate ligand concentrations (1 nM). A residual response apparent in Mel(1a) receptor-deficient C3H mice at higher melatonin concentrations (100 nM) is absent in Mel(1a)-Mel(1b) double-mutant mice, indicating that the Mel(1b) receptor mediates this effect of melatonin. These data indicate that there is a limited functional redundancy between the receptor subtypes in the SCN. Mice with targeted disruption of melatonin receptor subtypes will allow molecular dissection of other melatonin receptor-mediated responses. PMID- 12529410 TI - Inhibition of IkappaB kinase by a new class of retinoid-related anticancer agents that induce apoptosis. AB - The transcription factor NF-kappaB is overexpressed or constitutively activated in many cancer cells, where it induces expression of antiapoptotic genes correlating with resistance to anticancer therapies. Small molecules that inhibit the NF-kappaB signaling pathway could therefore be used to induce apoptosis in NF kappaB-overexpressing tumors and potentially serve as anticancer agents. We found that retinoid antagonist MX781 inhibited the activation of NF-kappaB-dependent transcriptional activity in different tumor cell lines. MX781 was able to completely inhibit tumor necrosis factor alpha-mediated activation of IkappaB kinase (IKK), the upstream regulator of NF-kappaB. Inhibition of IKK activity resulted from direct binding of MX781 to the kinase, as demonstrated by in vitro inhibition studies. Two other molecules, MX3350-1 and CD2325, which are retinoic acid receptor gamma-selective agonists, were capable of inhibiting IKK in vitro, although they exerted variable inhibition of IKK and NF-kappaB activities in intact cells in a cell type-specific manner. However, N-(4-hydroxyphenyl) retinamide, another apoptosis-inducing retinoid, and retinoic acid as well as other nonapoptotic retinoids did not inhibit IKK. Inhibition of IKK by the retinoid-related compounds and other small molecules correlated with reduced cell proliferation and increased apoptosis. Reduced cell viability was also observed after overexpression of an IKKbeta kinase-dead mutant or the IkappaBalpha superrepressor. The induction of apoptosis by the retinoid-related molecules that inhibited IKK was dependent on caspase activity but independent of the retinoid receptors. Thus, the presence of an excess of retinoic acid or a retinoid antagonist did not prevent the inhibition of IKK activation by MX781 and CD2325, indicating a retinoid receptor-independent mechanism of action. PMID- 12529411 TI - Essential role for NFI-C/CTF transcription-replication factor in tooth root development. AB - The mammalian tooth forms by a series of reciprocal epithelial-mesenchymal interactions. Although several signaling pathways and transcription factors have been implicated in regulating molar crown development, relatively little is known about the regulation of root development. Four genes encoding nuclear factor I (NFI) transcription-replication proteins are present in the mouse genome: Nfia, Nfib, Nfic, and NFIX: In order to elucidate its physiological role(s), we disrupted the Nfic gene in mice. Heterozygous animals appear normal, whereas Nfic(-/-) mice have unique tooth pathologies: molars lacking roots, thin and brittle mandibular incisors, and weakened abnormal maxillary incisors. Feeding in Nfic(-/-) mice is impaired, resulting in severe runting and premature death of mice reared on standard laboratory chow. However, a soft-dough diet mitigates the feeding impairment and maintains viability. Although Nfic is expressed in many organ systems, including the developing tooth, the tooth root development defects were the prominent phenotype. Indeed, molar crown development is normal, and well nourished Nfic(-/-) animals are fertile and can live as long as their wild-type littermates. The Nfic mutation is the first mutation described that affects primarily tooth root formation and should greatly aid our understanding of postnatal tooth development. PMID- 12529412 TI - Mitochondrial biogenesis and remodeling during adipogenesis and in response to the insulin sensitizer rosiglitazone. AB - White adipose tissue is an important endocrine organ involved in the control of whole-body metabolism, insulin sensitivity, and food intake. To better understand these functions, 3T3-L1 cell differentiation was studied by using combined proteomic and genomic strategies. The proteomics approach developed here exploits velocity gradient centrifugation as an alternative to isoelectric focusing for protein separation in the first dimension. A 20- to 30-fold increase in the concentration of numerous mitochondrial proteins was observed during adipogenesis, as determined by mass spectrometry and database correlation analysis. Light and electron microscopy confirmed a large increase in the number of mitochondrion profiles with differentiation. Furthermore, mRNA profiles obtained by using Affymetrix GeneChips revealed statistically significant increases in the expression of many nucleus-encoded mitochondrial genes during adipogenesis. Qualitative changes in mitochondrial composition also occur during adipose differentiation, as exemplified by increases in expression of proteins involved in fatty acid metabolism and of mitochondrial chaperones. Furthermore, the insulin sensitizer rosiglitazone caused striking changes in mitochondrial shape and expression of selective mitochondrial proteins. Thus, although mitochondrial biogenesis has classically been associated with brown adipocyte differentiation and thermogenesis, our results reveal that mitochondrial biogenesis and remodeling are inherent to adipose differentiation per se and are influenced by the actions of insulin sensitizers. PMID- 12529413 TI - Heregulin induces transcriptional activation of the progesterone receptor by a mechanism that requires functional ErbB-2 and mitogen-activated protein kinase activation in breast cancer cells. AB - The present study addresses the capacity of heregulin (HRG), a ligand of type I receptor tyrosine kinases, to transactivate the progesterone receptor (PR). For this purpose, we studied, on the one hand, an experimental model of hormonal carcinogenesis in which the synthetic progestin medroxyprogesterone acetate (MPA) induced mammary adenocarcinomas in female BALB/c mice and, on the other hand, the human breast cancer cell line T47D. HRG was able to exquisitely regulate biochemical attributes of PR in a way that mimicked PR activation by progestins. Thus, HRG treatment of primary cultures of epithelial cells of the progestin dependent C4HD murine mammary tumor line and of T47D cells induced a decrease of protein levels of PRA and -B isoforms and the downregulation of progesterone binding sites. HRG also promoted a significant increase in the percentage of PR localized in the nucleus in both cell types. DNA mobility shift assay revealed that HRG was able to induce PR binding to a progesterone response element (PRE) in C4HD and T47D cells. Transient transfections of C4HD and T47D cells with a plasmid containing a PRE upstream of a chloramphenicol acetyltransferase (CAT) gene demonstrated that HRG promoted a significant increase in CAT activity. In order to assess the molecular mechanisms underlying PR transactivation by HRG, we blocked ErbB-2 expression in C4HD and T47D cells by using antisense oligodeoxynucleotides to ErbB-2 mRNA, which resulted in the abolishment of HRG's capacity to induce PR binding to a PRE, as well as CAT activity in the transient transfection assays. Although the inhibition of HRG binding to ErbB-3 by an anti ErbB-3 monoclonal antibody suppressed HRG-induced PR activation, the abolishment of HRG binding to ErbB-4 had no effect on HRG activation of PR. To investigate the role of mitogen-activated protein kinases (MAPKs), we used the selective MEK1/MAPK inhibitor PD98059. Blockage of MAPK activation resulted in complete abrogation of HRG's capacity to induce PR binding to a PRE, as well as CAT activity. Finally, we demonstrate here for the first time that HRG-activated MAPK can phosphorylate both human and mouse PR in vitro. PMID- 12529414 TI - A Ral guanine exchange factor-Ral pathway is conserved in Drosophila melanogaster and sheds new light on the connectivity of the Ral, Ras, and Rap pathways. AB - Ras GTPases are central to many physiological and pathological signaling pathways and act via a combination of effectors. In mammals, at least three Ral exchange factors (RalGEFs) contain a Ras association domain and constitute a discrete subgroup of Ras effectors. Despite their ability to bind activated Rap as well as activated Ras, they seem to act downstream of Ras but not downstream of Rap. We have revisited the Ras/Rap-Ral connections in Drosophila melanogaster by using iterative two-hybrid screens with these three GTPases as primary baits and a subsequent genetic approach. We show that (i) the Ral-centered protein network appears to be extremely conserved in human and flies, (ii) in this network, RGL is a functional Drosophila orthologue of RalGEFs, and (iii) the RGL-Ral pathway functionally interacts with both the Ras and Rap pathways. Our data do not support the paradigmatic model where Ral is in the effector pathway of Ras. They reveal a signaling circuitry where Ral is functionally downstream of the Rap GTPase, at odds with the pathways described for mammalian cell lines. Thus, in vivo data show variations in the connectivity of pathways described for cell lines which might display only a subset of the biological possibilities. PMID- 12529416 TI - Identification and characterization of a novel human testis-specific Golgi protein, NYD-SP12. AB - A novel human testis-specific gene, NYD-SP12, was identified by hybridizing human adult or fetal testes cDNA samples with a human cDNA microarray containing 9216 clones. mRNA expression level of NYD-SP12 was 30-fold higher in human adult testes than fetal testes. Similarly, semi-quantitative RT-PCR revealed a differential expression pattern of an NYD-SP12 homologous gene in mouse adult and infant testes. PCR and hybridization analysis of NYD-SP12 mRNA from multiple human tissues indicated the expression of NYD-SP12 exclusively in the testis. In situ hybridization revealed that the expression of this gene was confined to spermatogenic epithelium and was not found in interstitial cells. NYD-SP12 transcript was not detected in patients with spermatogenic arrest and Sertoli cell-only syndrome. NYD-SP12 cDNA (GenBank accession number: AF345909) consisted of 2070 bp. The predicted 1707 bp open-reading fragment encoded a 569 amino acid protein that was 77% identical to a mouse homologue. Furthermore, computerized SMART and Motif analysis revealed that the protein contained a Structural Classification Of Proteins (SCOP) domain in the C-terminus and a cluster of phosphorylation sites for PKC, CK and cAMP/cGMP-dependent protein kinase. Interestingly, the EGFP-NYD-SP12 fusion protein was localized to the Golgi apparatus. In conclusion, the results suggest that NYD-SP12 is involved in spermatogenesis, and that NYD-SP12-encoded protein might function in the Golgi apparatus. PMID- 12529417 TI - Determination of the transcript profile of human endometrium. AB - The response of the human endometrium to the ovarian hormones, estrogen and progesterone, has been the focus of decades of research. In order to understand this critical aspect of endometrial physiology, we undertook a genome-wide analysis of transcript abundance and changes in transcript level between normal endometrium in the proliferative and secretory phases of the menstrual cycle. A high-density, oligonucleotide gene array, comprising 60 000 gene targets, was used to define the gene expression profile of proliferative and secretory phase endometrium. Results from the arrays were verified using real-time PCR. The expression levels of 149 transcripts differed significantly between the two phases of the cycle determined by stringent range limits (99.99%), calculated using local variance values. These transcripts include previously documented steroidally responsive genes (such as placental protein 14 and stromelysin-3) and novel transcripts not previously linked to either endometrial physiology or steroid regulation (such as intestinal trefoil factor and a number of expressed sequence tags). Examination of the 5' promoter regions of these genes identified many putative estrogen and progesterone receptor DNA binding domains, suggesting a direct response of these genes to the ovarian hormones. PMID- 12529415 TI - Interleukin-1beta induces glycosaminoglycan synthesis via the prostaglandin E2 pathway in cultured human cervical fibroblasts. AB - The aim of this study was to identify, in cultured human cervical fibroblasts, the mechanisms by which interleukin (IL)-1beta induces the synthesis of glycosaminoglycans (GAG) and to explore the putative role of prostaglandin E(2) (PGE(2)) in this process. Exposure of the cells for 24 h to IL-1beta induced a significant (P < 0.05) dose-dependent increase in GAG synthesis. IL-1beta (1 ng/ml) induced the expression of cyclooxygenase-2 (COX-2) protein 6 h after treatment, accompanied by a 7.5-fold increase in PGE(2) production. We confirmed that NS398, a selective COX-2 inhibitor, dose-dependently blocked PGE(2) augmentation following IL-1beta treatment. AH23848, the selective EP(4) receptor antagonist, completely abolished IL-1beta-induced GAG synthesis, whereas AH6809, an EP(2) receptor antagonist, had no effect on the stimulatory effects of IL 1beta. Furthermore, we demonstrated that 6 h exposure to IL-1beta induced a notable increase in EP(4) receptor mRNA expression and a decrease in EP(1) receptor mRNA but had no effect on the expression of EP(2) and EP(3) receptor transcripts. In conclusion, these findings indicate that IL-1beta not only induced GAG synthesis by increasing COX-2 protein expression and the subsequent PGE(2) production but also enhanced the responsiveness of cervical fibroblasts to PGE(2) by selectively up-regulating EP(4) receptor mRNA expression. These results suggest that PGE(2) may regulate human cervical ripening in an autocrine/paracrine manner via EP(4) receptors. PMID- 12529418 TI - The induction of baboon glycodelin expression by progesterone is not through Sp1. AB - Glycodelin is a major secretory product of the uterine glandular epithelial cells of the human and non-human primate during the late luteal phase of the menstrual cycle and early pregnancy. Since progesterone levels are elevated during these periods we sought to determine how progesterone modulates glycodelin gene expression. Co-transfection of various deletions of the baboon glycodelin promoter with the progesterone receptor (PR) into Ishikawa cells, a human endometrial cell line, revealed that full progesterone responsiveness is retained within the region -119/+48. In COS-1 cells, a kidney cell line, progesterone failed to elevate luciferase levels when various deletion constructs and the PR were co-transfected. Mutation of the Sp1 site in the -67/+48 region lowered basal expression but did not affect the ability of progesterone to increase expression of the luciferase reporter in Ishikawa cells. These findings suggest that Sp1 sites are not involved in the progesterone regulation of the baboon glycodelin gene. We propose that progesterone induces a factor that regulates glycodelin gene expression in the uterus since we failed to obtain a similar response in a non-uterine cell line. PMID- 12529419 TI - Leukocyte density and pro-inflammatory cytokine expression in human fetal membranes, decidua, cervix and myometrium before and during labour at term. AB - Accumulating evidence suggests that human parturition represents an inflammatory process. Leukocytes are known to infiltrate uterine tissues but the exact timing, nature and quantity of these cells has not been formally characterized. We have previously demonstrated an apparent increase in pro-inflammatory cytokines within tissues of the labouring uterus. The aims of this study were to quantify and compare the leukocyte subpopulations before and during labour in fetal membranes, decidua and cervix and to quantify and compare mRNA expression of interleukin 1beta (IL-1beta), IL-6, IL-8 and tumour necrosis factor-alpha in myometrium, cervix, chorio-decidua and amnion. Biopsies of each of these tissues were obtained from pregnant women delivered by Caesarean section before and after the onset of spontaneous labour at term. Subpopulations of leukocytes were identified using immunohistochemistry and cytokine mRNA expression was quantified using Northern analysis. We found that parturition was associated with a significant increase in IL-1beta, IL-6 and IL-8 mRNA expression in cervix and myometrium, IL 6 and IL-8 mRNA expression in chorio-decidua and IL-1beta and IL-8 mRNA expression in amnion. Histological analysis demonstrated that leukocytes (predominantly neutrophils and macrophages) infiltrate the uterine cervix coincident with the onset of labour. These data lend further support to the hypothesis that labour is an inflammatory process. PMID- 12529420 TI - Intercellular adhesion molecule-1 (ICAM-1) gene polymorphisms in endometriosis. AB - Endometriosis is a gynaecological disease with a certain genetic background, but the locations of possible genomic aberrations are still poorly clarified. Intercellular adhesion molecule-1 (ICAM-1), which is a surface glycoprotein that promotes adhesion in immunological and inflammatory reactions, seems to play a role in this condition. The aim of this study was to examine the potential associations of ICAM-1 gene polymorphisms with endometriosis and its severity. Specifically, we have studied two polymorphic sites located in codons 241 (G/R241) and 469 (E/K469) of the ICAM-1 gene. Three hundred and sixty-three Italian Caucasian women of reproductive age who underwent laparoscopy for benign pelvic conditions were enrolled in the study. Endometriosis was documented and staged in 188 women while 175 subjects, in whom endometriosis was laparoscopically ruled out, served as the control group. The frequency of the R241 allele was only marginally higher in endometriosis patients than in controls [5.8 versus 2.9%, P = 0.05; odds ratio (OR), 2.1; 95% confidence interval (CI), 1 4.5]. However, a strikingly high frequency of this allele was found in patients with Stage IV endometriosis versus controls (8.6 versus 2.8%, P = 0.008; OR, 3.2; 95% CI, 1.3-7.9). In contrast, the allele and genotype frequencies of the E/K469 polymorphism did not differ significantly between endometriosis and control groups. While the functional correlate of the G/R241 polymorphism remains unclear, this finding indicates that a genetic polymorphism in the ICAM-1 gene domain may contribute to the susceptibility to endometriosis. PMID- 12529421 TI - Interrelationships of interleukin-8 with interleukin-1beta and neutrophils in vaginal fluid of healthy and bacterial vaginosis positive women. AB - Vaginal innate immunity in response to microbial perturbation is still poorly understood and could be crucial for protection from adverse outcomes. We investigated the relationship between interleukin (IL)-8, IL-1beta and neutrophils in vaginal fluid obtained from 60 healthy women and 51 women who were bacterial vaginosis (BV) positive. Concentrations of IL-8 and IL-1beta were highly correlated with counts of neutrophils in vaginal fluid of the entire population examined (111 subjects). Vaginal IL-1beta concentrations were significantly higher (P < 0.001) in BV positive women. There was no significant difference in IL-8 levels or number of neutrophils between healthy controls and BV positive women. None of the healthy controls with high neutrophil counts (> or =75th percentile, 14 average count per field) had high concentrations of IL-1beta (> or =75th percentile, 220 pg/ml), whereas 84% of BV positive women with high neutrophil counts had high IL-1beta concentrations (P < 0.001). On the contrary, no difference in the percentage of subjects with elevated concentrations of IL-8 (> or =75th percentile, 2842 pg/ml) was found between healthy and BV positive women with high numbers of neutrophils (55.5% of healthy versus 53% of BV positive women). Our findings show that BV causes a large increase in IL-1beta concentrations which is not paralleled by an increase in IL-8 concentrations in vaginal fluid, suggesting that BV-associated factors more specifically dampen IL 8 rather than IL-1beta. The lack of an increase in IL-8 may explain the absence of an increase in neutrophil numbers in most women exposed to abnormal vaginal colonization (BV). PMID- 12529422 TI - N-terminal domain of yeast telomerase reverse transcriptase: recruitment of Est3p to the telomerase complex. AB - Telomerase is a reverse transcriptase that maintains chromosome ends. The N terminal half of the catalytic protein subunit (TERT) contains three functional domains (I, II, and III) that are conserved among TERTs but not found in other reverse transcriptases. Guided by an amino acid sequence alignment of nine TERT proteins, mutations were introduced into yeast TERT (Est2p). In support of the proposed alignment, mutation of virtually all conserved residues resulted in loss of-function or temperature sensitivity, accompanied by telomere shortening. Overexpression of telomerase component Est3p led to allele-specific suppression of the temperature-sensitive mutations in region I, suggesting that Est3p interacts with this protein domain. As predicted by the genetic results, a lethal mutation in region I resulted in loss of Est3p from the telomerase complex. We conclude that Est2p region I is required for the recruitment of Est3p to yeast telomerase. Given the phylogenetic conservation of region I of TERT, this protein domain may provide the equivalent function in all telomerases. PMID- 12529423 TI - The identification of pats1, a novel gene locus required for cytokinesis in Dictyostelium discoideum. AB - Here, we describe the identification and characterization of the cytokinesis deficient mutant cell line 17HG5, which was generated in a restriction enzyme mediated integration mutagenesis screen designed to isolate genes required for cytokinesis in Dictyostelium discoideum. Phenotypic characterization of the 17HG5 cell line revealed no apparent defects in the global functionality of the actomyosin cytoskeleton except for the observed cytokinesis defect when grown in suspension culture. Plasmid rescue was used to identify the disrupted gene locus (pats1; protein associated with the transduction of signal 1). that caused the cytokinesis defect. Disruption of the pats1 locus was recreated through homologous recombination in several independent cell lines, each recapitulating the cytokinesis-defective phenotype and thereby confirming that this gene locus is important for proper cytokinesis. Sequence data obtained by analysis of the genomic region flanking the inserted restriction enzyme-mediated integration plasmid revealed an 8892-bp genomic open reading frame encoding a 2964-amino-acid protein. The putative pats1 protein contains 3 regulatory domains (RI phosphatase, RII-GTP-binding, R-III protein kinase), 13 leucine-rich repeats, and 8 WD-40 repeats. These regulatory domains coupled with the protein-protein interacting domains suggest that pats1 is involved in signal transduction during cytokinesis in Dictyostelium. PMID- 12529424 TI - A Bni4-Glc7 phosphatase complex that recruits chitin synthase to the site of bud emergence. AB - Bni4 is a scaffold protein in the yeast Saccharomyces cerevisiae that tethers chitin synthase III to the bud neck by interacting with septin neck filaments and with Chs4, a regulatory subunit of chitin synthase III. We show herein that Bni4 is also a limiting determinant for the targeting of the type 1 serine/threonine phosphatase (Glc7) to the bud neck. Yeast cells containing a Bni4 variant that fails to associate with Glc7 fail to tether Chs4 to the neck, due in part to the failure of Bni4(V831A/F833A) to localize properly. Conversely, the Glc7-129 mutant protein fails to bind Bni4 properly and glc7-129 mutants exhibit reduced levels of Bni4 at the bud neck. Bni4 is phosphorylated in a cell cycle-dependent manner and Bni4(V831A/F833A) is both hyperphosphorylated and mislocalized in vivo. Yeast cells lacking the protein kinase Hsl1 exhibit increased levels of Bni4-GFP at the bud neck. GFP-Chs4 does not accumulate at the incipient bud site in either a bni4::TRP1 or a bni4(V831A/F833A) mutant but does mobilize to the neck at cytokinesis. Together, these results indicate that the formation of the Bni4-Glc7 complex is required for localization to the site of bud emergence and for subsequent targeting of chitin synthase. PMID- 12529425 TI - STAT5a activation mediates the epithelial to mesenchymal transition induced by oncogenic RhoA. AB - The involvement of Rho GTPases in signal transduction pathways leading to transcription activation is one of the major roles of this family of GTPases. Thus, the identification of transcription factors regulated by Rho GTPases and the understanding of the mechanisms of their activation and its biological outcome are of great interest. Here, we provide evidence that Rho GTPases modulate Stat5a, a transcription factor of the family of signal transducers and activators of transcription. RhoA triggers tyrosine phosphorylation (Y696) of Stat5a via a JAK2-dependent mechanism and promotes DNA-binding activity of Stat5a. Tyrosine phosphorylation of Stat5a is also stimulated physiologically by lysophosphatidic acid (LPA) in a Rho-dependent manner. Simultaneously, RhoA reduces serine phosphorylation of Stat5a at both serine residues S726 and S780, resulting in a further increase of activity as defined by mutagenesis experiments. Furthermore, serine dephosphorylation of Stat5a by RhoA does not take place by down-modulation of either JNK1, MEK1, or p38 MAP kinases, as determined by transfection experiments or chemical inhibition of both MEK1, p38, and JNK serine kinases. Thus, RhoA regulates Stat5a via tyrosine phosphorylation and via a yet to be determined novel down-modulating pathway that involves serine dephosphorylation. Finally, we provide evidence for a role of Stat5a in RhoA induced epithelial-to-mesenchymal transition with concomitant increase in vimentin expression, E-cadherin down-regulation, and cell motility. PMID- 12529426 TI - Rapid up-regulation of alpha4 integrin-mediated leukocyte adhesion by transforming growth factor-beta1. AB - The alpha4 integrins (alpha4beta1 and alpha4beta7) are cell surface heterodimers expressed mostly on leukocytes that mediate cell-cell and cell-extracellular matrix adhesion. A characteristic feature of alpha4 integrins is that their adhesive activity can be subjected to rapid modulation during the process of cell migration. Herein, we show that transforming growth factor-beta1 (TGF-beta1) rapidly (0.5-5 min) and transiently up-regulated alpha4 integrin-dependent adhesion of different human leukocyte cell lines and human peripheral blood lymphocytes (PBLs) to their ligands vascular cell adhesion molecule-1 (VCAM-1) and connecting segment-1/fibronectin. In addition, TGF-beta1 enhanced the alpha4 integrin-mediated adhesion of PBLs to tumor necrosis factor-alpha-treated human umbilical vein endothelial cells, indicating the stimulation of alpha4beta1/VCAM 1 interaction. Although TGF-beta1 rapidly activated the small GTPase RhoA and the p38 mitogen-activated protein kinase, enhanced adhesion did not require activation of both signaling molecules. Instead, polymerization of actin cytoskeleton triggered by TGF-beta1 was necessary for alpha4 integrin-dependent up-regulated adhesion, and elevation of intracellular cAMP opposed this up regulation. Moreover, TGF-beta1 further increased cell adhesion mediated by alpha4 integrins in response to the chemokine stromal cell-derived factor-1alpha. These data suggest that TGF-beta1 can potentially contribute to cell migration by dynamically regulating cell adhesion mediated by alpha4 integrins. PMID- 12529427 TI - Caspase- and serine protease-dependent apoptosis by the death domain of FADD in normal epithelial cells. AB - The adapter protein FADD consists of two protein interaction domains: a death domain and a death effector domain. The death domain binds to activated death receptors such as Fas, whereas the death effector domain binds to procaspase 8. An FADD mutant, which consists of only the death domain (FADD-DD), inhibits death receptor-induced apoptosis. FADD-DD can also activate a mechanistically distinct, cell type-specific apoptotic pathway that kills normal but not cancerous prostate epithelial cells. Here, we show that this apoptosis occurs through activation of caspases 9, 3, 6, and 7 and a serine protease. Simultaneous inhibition of caspases and serine proteases prevents FADD-DD-induced death. Inhibition of either pathway alone does not prevent cell death but does affect the morphology of the dying cells. Normal prostate epithelial cells require both the caspase and serine protease inhibitors to efficiently prevent apoptosis in response to TRAIL. In contrast, the serine protease inhibitor does not affect TRAIL-induced death in prostate tumor cells suggesting that the FADD-DD-dependent pathway can be activated by TRAIL. This apoptosis pathway is activated in a cell type-specific manner that is defective in cancer cells, suggesting that this pathway may be targeted during cancer development. PMID- 12529428 TI - Localization, dynamics, and function of survivin revealed by expression of functional survivinDsRed fusion proteins in the living cell. AB - Survivin, a member of the inhibitor of apoptosis protein family, has attracted growing attention due to its expression in various tumors and its potential application in tumor therapy. However, its subcellular localization and function have remained controversial: Recent studies revealed that survivin is localized at the mitotic spindle, binds caspases, and could thus protect cells from apoptosis. The cell cycle-dependent expression of survivin and its antiapoptotic function led to the hypothesis that survivin connects the cell cycle with apoptosis, thus providing a death switch for the termination of defective mitosis. In other studies, survivin was detected at kinetochores, cleavage furrow, and midbody, localizations being characteristic for chromosomal passenger proteins. These proteins are involved in cytokinesis as inferred from the observation that RNA interference and expression of mutant proteins led to cytokinesis defects without an increase in apoptosis. To remedy these discrepancies, we analyzed the localizations of a survivinDsRed fusion protein in HeLa cells by using confocal laser scanning microscopy and time-lapse video imaging. SurvivinDsRed was excluded from the interphase nucleus and was detected in centrosomes and at kinetochores. It dissociated from chromosomes at the anaphase/telophase transition and accumulated at the ends of polar microtubuli where it was immediately condensed to the midbody. Overexpression of both survivinDsRed and of a phosphorylation-defective mutant conferred resistance against apoptosis-inducing reagents, but only the overexpressed mutant protein caused an aberrant cytokinesis. These data characterize in detail the dynamics of survivin in vertebrate cells and confirm that survivin represents a chromosomal passenger protein. PMID- 12529429 TI - Long-term potentiation of exocytosis and cell membrane repair in fibroblasts. AB - We previously found that a microdisruption of the plasma membrane evokes Ca(2+) regulated exocytosis near the wound site, which is essential for membrane resealing. We demonstrate herein that repeated membrane disruption reveals long term potentiation of Ca(2+)-regulated exocytosis in 3T3 fibroblasts, which is closely correlated with faster membrane resealing rates. This potentiation of exocytosis is cAMP-dependent protein kinase A dependent in the early stages (minutes), in the intermediate term (hours) requires protein synthesis, and for long term (24 h) depends on the activation of cAMP response element-binding protein (CREB). We were able to demonstrate that wounding cells activated CREB within 3.5 h. In all three phases, the increase in the amount of exocytosis was correlated with an increase in the rate of membrane resealing. However, a brief treatment with forskolin, which is effective for short-term potentiation and which could also activate CREB, was not sufficient to induce long-term potentiation of resealing. These results imply that long-term potentiation by CREB required activation by another, cAMP-independent pathway. PMID- 12529430 TI - Time-lapse imaging reveals dynamic relocalization of PP1gamma throughout the mammalian cell cycle. AB - Protein phosphatase 1 (PP1) is a ubiquitous serine/threonine phosphatase that regulates many cellular processes, including cell division. When transiently expressed as fluorescent protein (FP) fusions, the three PP1 isoforms, alpha, beta/delta, and gamma1, are active phosphatases with distinct localization patterns. We report here the establishment and characterization of HeLa cell lines stably expressing either FP-PP1gamma or FP alone. Time-lapse imaging reveals dynamic targeting of FP-PP1gamma to specific sites throughout the cell cycle, contrasting with the diffuse pattern observed for FP alone. FP-PP1gamma shows a nucleolar accumulation during interphase. On entry into mitosis, it localizes initially at kinetochores, where it exchanges rapidly with the diffuse cytoplasmic pool. A dramatic relocalization of PP1 to the chromosome-containing regions occurs at the transition from early to late anaphase, and by telophase FP PP1gamma also accumulates at the cleavage furrow and midbody. The changing spatio temporal distribution of PP1gamma revealed using the stable PP1 cell lines implicates it in multiple processes, including nucleolar function, the regulation of chromosome segregation and cytokinesis. PMID- 12529431 TI - A balance of capping protein and profilin functions is required to regulate actin polymerization in Drosophila bristle. AB - Profilin is a well-characterized protein known to be important for regulating actin filament assembly. Relatively few studies have addressed how profilin interacts with other actin-binding proteins in vivo to regulate assembly of complex actin structures. To investigate the function of profilin in the context of a differentiating cell, we have studied an instructive genetic interaction between mutations in profilin (chickadee) and capping protein (cpb). Capping protein is the principal protein in cells that caps actin filament barbed ends. When its function is reduced in the Drosophila bristle, F-actin levels increase and the actin cytoskeleton becomes disorganized, causing abnormal bristle morphology. chickadee mutations suppress the abnormal bristle phenotype and associated abnormalities of the actin cytoskeleton seen in cpb mutants. Furthermore, overexpression of profilin in the bristle mimics many features of the cpb loss-of-function phenotype. The interaction between cpb and chickadee suggests that profilin promotes actin assembly in the bristle and that a balance between capping protein and profilin activities is important for the proper regulation of F-actin levels. Furthermore, this balance of activities affects the association of actin structures with the membrane, suggesting a link between actin filament dynamics and localization of actin structures within the cell. PMID- 12529432 TI - Piecemeal microautophagy of nucleus in Saccharomyces cerevisiae. AB - Nucleus-vacuole (NV) junctions in Saccharomyces cerevisiae are formed through specific interactions between Vac8p on the vacuole membrane and Nvj1p in the nuclear envelope. Herein, we report that NV junctions in yeast promote piecemeal microautophagy of the nucleus (PMN). During PMN, teardrop-like blebs are pinched from the nucleus, released into the vacuole lumen, and degraded by soluble hydrolases. PMN occurs in rapidly dividing cells but is induced to higher levels by carbon and nitrogen starvation and is under the control of the Tor kinase nutrient-sensing pathway. Confocal and biochemical assays demonstrate that Nvj1p is degraded in a PMN-dependent manner. PMN occurs normally in apg7-delta cells and is, therefore, not dependent on macroautophagy. Transmission electron microscopy reveals that portions of the granular nucleolus are often sequestered into PMN structures. These results introduce a novel mode of selective microautophagy that targets nonessential components of the yeast nucleus for degradation and recycling in the vacuole. PMID- 12529433 TI - Visualization of TGN to endosome trafficking through fluorescently labeled MPR and AP-1 in living cells. AB - We have stably expressed in HeLa cells a chimeric protein made of the green fluorescent protein (GFP) fused to the transmembrane and cytoplasmic domains of the mannose 6-phosphate/insulin like growth factor II receptor in order to study its dynamics in living cells. At steady state, the bulk of this chimeric protein (GFP-CI-MPR) localizes to the trans-Golgi network (TGN), but significant amounts are also detected in peripheral, tubulo-vesicular structures and early endosomes as well as at the plasma membrane. Time-lapse videomicroscopy shows that the GFP CI-MPR is ubiquitously detected in tubular elements that detach from the TGN and move toward the cell periphery, sometimes breaking into smaller tubular fragments. The formation of the TGN-derived tubules is temperature dependent, requires the presence of intact microtubule and actin networks, and is regulated by the ARF-1 GTPase. The TGN-derived tubules fuse with peripheral, tubulo vesicular structures also containing the GFP-CI-MPR. These structures are highly dynamic, fusing with each other as well as with early endosomes. Time-lapse videomicroscopy performed on HeLa cells coexpressing the CFP-CI-MPR and the AP-1 complex whose gamma-subunit was fused to YFP shows that AP-1 is present not only on the TGN and peripheral CFP-CI-MPR containing structures but also on TGN derived tubules containing the CFP-CI-MPR. The data support the notion that tubular elements can mediate MPR transport from the TGN to a peripheral, tubulo vesicular network dynamically connected with the endocytic pathway and that the AP-1 coat may facilitate MPR sorting in the TGN and endosomes. PMID- 12529434 TI - A new identity for MLK3 as an NIMA-related, cell cycle-regulated kinase that is localized near centrosomes and influences microtubule organization. AB - Although conserved counterparts for most proteins involved in the G(2)/M transition of the cell cycle have been found in all eukaryotes, a notable exception is the essential but functionally enigmatic fungal kinase NIMA. While a number of vertebrate kinases have been identified with catalytic domain homology to NIMA, none of these resemble NIMA within its extensive noncatalytic region, a region critical for NIMA function in Aspergillus nidulans. We used a bioinformatics approach to search for proteins with homology to the noncatalytic region of NIMA and identified mixed lineage kinase 3 (MLK3). MLK3 has been proposed to serve as a component in MAP kinase cascades, particularly those resulting in the activation of the c-Jun N-terminal kinase (JNK). Here we describe the first in-depth study of endogenous MLK3 and report that, like NIMA, MLK3 phosphorylation and activity are enhanced during G(2)/M, whereas JNK remains inactive. Coincident with the G(2)/M transition, a period marked by dramatic reorganization of the cytoplasmic microtubule network, endogenous MLK3 transiently disperses away from the centrosome and centrosomal-proximal sites where it is localized during interphase. Furthermore, when overexpressed, MLK3, like NIMA, localizes to the centrosomal region, induces profound disruption of cytoplasmic microtubules and a nuclear distortion phenotype that differs from mitotic chromosome condensation. Cellular depletion of MLK3 protein using siRNA technology results in an increased sensitivity to the microtubule-stabilizing agent taxol. Our studies suggest a new role for MLK3, separable from its function in the JNK pathway, that may contribute to promoting microtubule instability, a hallmark of M phase entry. PMID- 12529436 TI - The Drosophila Cog5 homologue is required for cytokinesis, cell elongation, and assembly of specialized Golgi architecture during spermatogenesis. AB - The multisubunit conserved oligomeric Golgi (COG) complex has been shown previously to be involved in Golgi function in yeast and mammalian tissue culture cells. Despite this broad conservation, several subunits, including Cog5, were not essential for growth and showed only mild effects on secretion when mutated in yeast, raising questions about what functions these COG complex subunits play in the life of the cell. Here, we show that function of the gene four way stop (fws), which encodes the Drosophila Cog5 homologue, is necessary for dramatic changes in cellular and subcellular morphology during spermatogenesis. Loss-of function mutations in fws caused failure of cleavage furrow ingression in dividing spermatocytes and failure of cell elongation in differentiating spermatids and disrupted the formation and/or stability of the Golgi-based spermatid acroblast. Consistent with the lack of a growth defect in yeast lacking Cog5, animals lacking fws function were viable, although males were sterile. Fws protein localized to Golgi structures throughout spermatogenesis. We propose that Fws may directly or indirectly facilitate efficient vesicle traffic through the Golgi to support rapid and extensive increases in cell surface area during spermatocyte cytokinesis and polarized elongation of differentiating spermatids. Our study suggests that Drosophila spermatogenesis can be an effective sensitized genetic system to uncover in vivo functions for proteins involved in Golgi architecture and/or vesicle transport. PMID- 12529435 TI - Osteopontin deficiency produces osteoclast dysfunction due to reduced CD44 surface expression. AB - Osteopontin (OPN) was expressed in murine wild-type osteoclasts, localized to the basolateral, clear zone, and ruffled border membranes, and deposited in the resorption pits during bone resorption. The lack of OPN secretion into the resorption bay of avian osteoclasts may be a component of their functional resorption deficiency in vitro. Osteoclasts deficient in OPN were hypomotile and exhibited decreased capacity for bone resorption in vitro. OPN stimulated CD44 expression on the osteoclast surface, and CD44 was shown to be required for osteoclast motility and bone resorption. Exogenous addition of OPN to OPN-/- osteoclasts increased the surface expression of CD44, and it rescued osteoclast motility due to activation of the alpha(v)beta(3) integrin. Exogenous OPN only partially restored bone resorption because addition of OPN failed to produce OPN secretion into resorption bays as seen in wild-type osteoclasts. As expected with these in vitro findings of osteoclast dysfunction, a bone phenotype, heretofore unappreciated, was characterized in OPN-deficient mice. Delayed bone resorption in metaphyseal trabeculae and diminished eroded perimeters despite an increase in osteoclast number were observed in histomorphometric measurements of tibiae isolated from OPN-deficient mice. The histomorphometric findings correlated with an increase in bone rigidity and moment of inertia revealed by load-to-failure testing of femurs. These findings demonstrate the role of OPN in osteoclast function and the requirement for OPN as an osteoclast autocrine factor during bone remodeling. PMID- 12529438 TI - Global transcriptional responses of fission yeast to environmental stress. AB - We explored transcriptional responses of the fission yeast Schizosaccharomyces pombe to various environmental stresses. DNA microarrays were used to characterize changes in expression profiles of all known and predicted genes in response to five stress conditions: oxidative stress caused by hydrogen peroxide, heavy metal stress caused by cadmium, heat shock caused by temperature increase to 39 degrees C, osmotic stress caused by sorbitol, and DNA damage caused by the alkylating agent methylmethane sulfonate. We define a core environmental stress response (CESR) common to all, or most, stresses. There was a substantial overlap between CESR genes of fission yeast and the genes of budding yeast that are stereotypically regulated during stress. CESR genes were controlled primarily by the stress-activated mitogen-activated protein kinase Sty1p and the transcription factor Atf1p. S. pombe also activated gene expression programs more specialized for a given stress or a subset of stresses. In general, these "stress-specific" responses were less dependent on the Sty1p mitogen-activated protein kinase pathway and may involve specific regulatory factors. Promoter motifs associated with some of the groups of coregulated genes were identified. We compare and contrast global regulation of stress genes in fission and budding yeasts and discuss evolutionary implications. PMID- 12529437 TI - CRM1/Ran-mediated nuclear export of p27(Kip1) involves a nuclear export signal and links p27 export and proteolysis. AB - We show that p27 localization is cell cycle regulated and we suggest that active CRM1/RanGTP-mediated nuclear export of p27 may be linked to cytoplasmic p27 proteolysis in early G1. p27 is nuclear in G0 and early G1 and appears transiently in the cytoplasm at the G1/S transition. Association of p27 with the exportin CRM1 was minimal in G0 and increased markedly during G1-to-S phase progression. Proteasome inhibition in mid-G1 did not impair nuclear import of p27, but led to accumulation of p27 in the cytoplasm, suggesting that export precedes degradation for at least part of the cellular p27 pool. p27-CRM1 binding and nuclear export were inhibited by S10A mutation but not by T187A mutation. A putative nuclear export sequence in p27 is identified whose mutation reduced p27 CRM1 interaction, nuclear export, and p27 degradation. Leptomycin B (LMB) did not inhibit p27-CRM1 binding, nor did it prevent p27 export in vitro or in heterokaryon assays. Prebinding of CRM1 to the HIV-1 Rev nuclear export sequence did not inhibit p27-CRM1 interaction, suggesting that p27 binds CRM1 at a non-LMB sensitive motif. LMB increased total cellular p27 and may do so indirectly, through effects on other p27 regulatory proteins. These data suggest a model in which p27 undergoes active, CRM1-dependent nuclear export and cytoplasmic degradation in early G1. This would permit the incremental activation of cyclin E Cdk2 leading to cyclin E-Cdk2-mediated T187 phosphorylation and p27 proteolysis in late G1 and S phase. PMID- 12529439 TI - The protein tyrosine phosphatase PTP-BL associates with the midbody and is involved in the regulation of cytokinesis. AB - PTP-BL is a highly modular protein tyrosine phosphatase of unknown function. It consists of an N-terminal FERM domain, five PDZ domains, and a C-terminally located tyrosine phosphatase domain. Here we show that PTP-BL is involved in the regulation of cytokinesis. We demonstrate localization of endogenous PTP-BL at the centrosomes during inter- and metaphase and at the spindle midzone during anaphase. Finally PTP-BL is concentrated at the midbody in cytokinesis. We show that PTP-BL is targeted to the midbody and centrosome by a specific splicing variant of the N-terminus characterized by an insertion of 182 amino acids. Moreover, we demonstrate that the FERM domain of PTP-BL is associated with the contractile ring and can be cosedimented with filamentous actin, whereas the N terminus can be cosedimented with microtubules. We demonstrate that elevating the expression level of wild-type PTP-BL or expression of PTP-BL with an inactive tyrosine phosphatase domain leads to defects in cytokinesis and to the generation of multinucleate cells. We suggest that PTP-BL plays a role in the regulation of cytokinesis. PMID- 12529440 TI - Deregulation of the Egfr/Ras signaling pathway induces age-related brain degeneration in the Drosophila mutant vap. AB - Ras signaling has been shown to play an important role in promoting cell survival in many different tissues. Here we show that upregulation of Ras activity in adult Drosophila neurons induces neuronal cell death, as evident from the phenotype of vacuolar peduncle (vap) mutants defective in the Drosophila RasGAP gene, which encodes a Ras GTPase-activating protein. These mutants show age related brain degeneration that is dependent on activation of the EGF receptor signaling pathway in adult neurons, leading to autophagic cell death (cell death type 2). These results provide the first evidence for a requirement of Egf receptor activity in differentiated adult Drosophila neurons and show that a delicate balance of Ras activity is essential for the survival of adult neurons. PMID- 12529441 TI - Cloning, localization, and axonemal function of Tetrahymena centrin. AB - Centrin, an EF hand Ca(2+) binding protein, has been cloned in Tetrahymena thermophila. It is a 167 amino acid protein of 19.4 kDa with a unique N-terminal region, coded by a single gene containing an 85-base pair intron. It has > 80% homology to other centrins and high homology to Tetrahymena EF hand proteins calmodulin, TCBP23, and TCBP25. Specific cellular localizations of the closely related Tetrahymena EF hand proteins are different from centrin. Centrin is localized to basal bodies, cortical fibers in oral apparatus and ciliary rootlets, the apical filament ring and to inner arm (14S) dynein (IAD) along the ciliary axoneme. The function of centrin in Ca(2+) control of IAD activity was explored using in vitro microtubule (MT) motility assays. Ca(2+) or the Ca(2+) mimicking peptide CALP1, which binds EF hand proteins in the absence of Ca(2+), increased MT sliding velocity. Antibodies to centrin abrogated this increase. This is the first demonstration of a specific centrin function associated with axonemal dynein. It suggests that centrin is a key regulatory protein for Tetrahymena axonemal Ca(2+) responses, including ciliary reversal or chemotaxis. PMID- 12529442 TI - SVIP is a novel VCP/p97-interacting protein whose expression causes cell vacuolation. AB - VCP/p97 is involved in a variety of cellular processes, including membrane fusion and ubiquitin-dependent protein degradation. It has been suggested that adaptor proteins such as p47 and Ufd1p confer functional versatility to VCP/p97. To identify novel adaptors, we searched for proteins that interact specifically with VCP/p97 by using the yeast two-hybrid system, and discovered a novel VCP/p97 interacting protein named small VCP/p97-interacting protein (SVIP). Rat SVIP is a 76-amino acid protein that contains two putative coiled-coil regions, and potential myristoylation and palmitoylation sites at the N terminus. Binding experiments revealed that the N-terminal coiled-coil region of SVIP, and the N terminal and subsequent ATP-binding regions (ND1 domain) of VCP/p97, interact with each other. SVIP and previously identified adaptors p47 and ufd1p interact with VCP/p97 in a mutually exclusive manner. Overexpression of full-length SVIP or a truncated mutant did not markedly affect the structure of the Golgi apparatus, but caused extensive cell vacuolation reminiscent of that seen upon the expression of VCP/p97 mutants or polyglutamine proteins in neuronal cells. The vacuoles seemed to be derived from endoplasmic reticulum membranes. These results together suggest that SVIP is a novel VCP/p97 adaptor whose function is related to the integrity of the endoplasmic reticulum. PMID- 12529443 TI - Sam68 RNA binding protein is an in vivo substrate for protein arginine N methyltransferase 1. AB - RNA binding proteins often contain multiple arginine glycine repeats, a sequence that is frequently methylated by protein arginine methyltransferases. The role of this posttranslational modification in the life cycle of RNA binding proteins is not well understood. Herein, we report that Sam68, a heteronuclear ribonucleoprotein K homology domain containing RNA binding protein, associates with and is methylated in vivo by the protein arginine N-methyltransferase 1 (PRMT1). Sam68 contains asymmetrical dimethylarginines near its proline motif P3 as assessed by using a novel asymmetrical dimethylarginine-specific antibody and mass spectrometry. Deletion of the methylation sites and the use of methylase inhibitors resulted in Sam68 accumulation in the cytoplasm. Sam68 was also detected in the cytoplasm of PRMT1-deficient embryonic stem cells. Although the cellular function of Sam68 is unknown, it has been shown to export unspliced human immunodeficiency virus RNAs. Cells treated with methylase inhibitors prevented the ability of Sam68 to export unspliced human immunodeficiency virus RNAs. Other K homology domain RNA binding proteins, including SLM-1, SLM-2, QKI 5, GRP33, and heteronuclear ribonucleoprotein K were also methylated in vivo. These findings demonstrate that RNA binding proteins are in vivo substrates for PRMT1, and their methylation is essential for their proper localization and function. PMID- 12529444 TI - Cell cycle-dependent regulation of structure of endoplasmic reticulum and inositol 1,4,5-trisphosphate-induced Ca2+ release in mouse oocytes and embryos. AB - The organization of endoplasmic reticulum (ER) was examined in mouse eggs undergoing fertilization and in embryos during the first cell cycle. The ER in meiosis II (MII)-arrested mouse eggs is characterized by accumulations (clusters) that are restricted to the cortex of the vegetal hemisphere of the egg. Monitoring ER structure with DiI18 after egg activation has demonstrated that ER clusters disappear at the completion of meiosis II. The ER clusters can be maintained by inhibiting the decrease in cdk1-cyclin B activity by using the proteasome inhibitor MG132, or by microinjecting excess cyclin B. A role for cdk1 cyclin B in ER organization is further suggested by the finding that the cdk inhibitor roscovitine causes the loss of ER clusters in MII eggs. Cortical clusters are specific to meiosis as they do not return in the first mitotic division; rather, the ER aggregates around the mitotic spindle. Inositol 1,4,5 trisphosphate-induced Ca(2+) release is also regulated in a cell cycle-dependent manner where it is increased in MII and in the first mitosis. The cell cycle dependent effects on ER structure and inositol 1,4,5-trisphosphate-induced Ca(2+) release have implications for understanding meiotic and mitotic control of ER structure and inheritance, and of the mechanisms regulating mitotic Ca(2+) signaling. PMID- 12529445 TI - Yeast glycogen synthase kinase-3 activates Msn2p-dependent transcription of stress responsive genes. AB - The yeast Saccharomyces cerevisiae has four genes, MCK1, MDS1 (RIM11), MRK1, and YOL128c, that encode homologues of mammalian glycogen synthase kinase 3 (GSK-3). A gsk-3 null mutant in which these four genes are disrupted showed growth defects on galactose medium. We isolated several multicopy suppressors of this growth defect. Two of them encoded Msn2p and phosphoglucomutase (PGM). Msn2p is a transcription factor that binds to the stress-response element (STRE). PGM is an enzyme that interconverts glucose-1 phosphate and glucose-6 phosphate and is regulated by Msn2p at the transcriptional level. Expression of the mRNAs of PGM2 and DDR2, whose promoter regions possess STRE sequences, on induction by heat shock or salt stress was reduced not only in an msn2 msn4 (msn2 homologue) double mutant but also in the gsk-3 null mutant. STRE-dependent transcription was greatly inhibited in the gsk-3 null mutant or mck1 mds1 double mutant, and this phenotype was suppressed by the expression of Mck1p but not of a kinase-inactive form of Mck1p. Although Msn2p accumulated in the nucleus of the gsk-3 null mutant as well as in the wild-type strain under various stress conditions, its STRE binding activity was reduced in extracts prepared from the gsk-3 null mutant or mck1 mds1 double mutant. These results suggest that yeast GSK-3 promotes formation of a complex between Msn2p and DNA, which is required for the proper response to different forms of stress. Because neither Msn2p-GSK-3 complex formation nor GSK-3-dependent phosphorylation of Msn2p could be detected, the regulation of Msn2p by GSK-3 may be indirect. PMID- 12529446 TI - Gef1p, a new guanine nucleotide exchange factor for Cdc42p, regulates polarity in Schizosaccharomyces pombe. AB - Schizosaccharomyces pombe cdc42(+) regulates cell morphology and polarization of the actin cytoskeleton. Scd1p/Ral1p is the only described guanine nucleotide exchange factor (GEF) for Cdc42p in S. pombe. We have identified a new GEF, named Gef1p, specifically regulating Cdc42p. Gef1p binds to inactive Cdc42p but not to other Rho GTPases in two-hybrid assays. Overexpression of gef1(+) increases specifically the GTP-bound Cdc42p, and Gef1p is capable of stimulating guanine nucleotide exchange of Cdc42p in vitro. Overexpression of gef1(+) causes changes in cell morphology similar to those caused by overexpression of the constitutively active cdc42G12V allele. Gef1p localizes to the septum. gef1(+) deletion is viable but causes a mild cell elongation and defects in bipolar growth and septum formation, suggesting a role for Gef1p in the control of cell polarity and cytokinesis. The double mutant gef1delta scd1delta is not viable, indicating that they share an essential function as Cdc42p activators. However, both deletion and overexpression of either gef1(+) or scd1(+) causes different morphological phenotypes, which suggest different functions. Genetic evidence revealed a link between Gef1p and the signaling pathway of Shk1/Orb2p and Orb6p. In contrast, no genetic interaction between Gef1p and Shk2p-Mkh1p pathway was observed. PMID- 12529447 TI - Interactions among COX1, COX2, and COX3 mRNA-specific translational activator proteins on the inner surface of the mitochondrial inner membrane of Saccharomyces cerevisiae. AB - The core of the cytochrome c oxidase complex is composed of its three largest subunits, Cox1p, Cox2p, and Cox3p, which are encoded in mitochondrial DNA of Saccharomyces cerevisiae and inserted into the inner membrane from the inside. Mitochondrial translation of the COX1, COX2, and COX3 mRNAs is activated mRNA specifically by the nuclearly coded proteins Pet309p, Pet111p, and the concerted action of Pet54p, Pet122p, and Pet494p, respectively. Because the translational activators recognize sites in the 5'-untranslated leaders of these mRNAs and because untranslated mRNA sequences contain information for targeting their protein products, the activators are likely to play a role in localizing translation. Herein, we report physical associations among the mRNA-specific translational activator proteins, located on the matrix side of the inner membrane. These interactions, detected by coimmune precipitation and by two hybrid experiments, suggest that the translational activator proteins could be organized on the surface of the inner membrane such that synthesis of Cox1p, Cox2p, and Cox3p would be colocalized in a way that facilitates assembly of the core of the cytochrome c oxidase complex. In addition, we found interactions between Nam1p/Mtf2p and the translational activators, suggesting an organized delivery of mitochondrial mRNAs to the translation system. PMID- 12529449 TI - Mild cognitive impairment: a clinically useful but currently ill-defined concept? AB - The term mild cognitive impairment (MCI) has gained wide currency among clinicians, and particularly dementia researchers, to denote patients with memory deficits who do not yet fulfil the criteria for dementia, but are at high risk of conversion. MCI is therefore regarded as the prodromic or pre-dementia stage of Alzheimer's disease. The accurate categorization of these subjects has far reaching implications, both for research in this field and for those individuals who fall within this diagnostic group. Despite a wealth of studies examining the neuropsychological, neuroimaging and biological profiles of this population, the characterization of MCI remains controversial. This brief overview discusses a number of the issues related to this topic and questions the currently accepted criteria. PMID- 12529448 TI - Regulation of vascular endothelial growth factor receptor-2 activity by caveolin 1 and plasma membrane cholesterol. AB - The stimulation of vascular endothelial growth factor receptor-2 (VEGFR-2) by tumor-derived VEGF represents a key event in the initiation of angiogenesis. In this work, we report that VEGFR-2 is localized in endothelial caveolae, associated with caveolin-1, and that this complex is rapidly dissociated upon stimulation with VEGF. The kinetics of caveolin-1 dissociation correlated with those of VEGF-dependent VEGFR-2 tyrosine phosphorylation, suggesting that caveolin-1 acts as a negative regulator of VEGF R-2 activity. Interestingly, we observed that in an overexpression system in which VEGFR-2 is constitutively active, caveolin-1 overexpression inhibits VEGFR-2 activity but allows VEGFR-2 to undergo VEGF-dependent activation, suggesting that caveolin-1 can confer ligand dependency to a receptor system. Removal of caveolin and VEGFR-2 from caveolae by cholesterol depletion resulted in an increase in both basal and VEGF-induced phosphorylation of VEGFR-2, but led to the inhibition of VEGF-induced ERK activation and endothelial cell migration, suggesting that localization of VEGFR 2 to these domains is crucial for VEGF-mediated signaling. Dissociation of the VEGFR-2/caveolin-1 complex by VEGF or cyclodextrin led to a PP2-sensitive phosphorylation of caveolin-1 on tyrosine 14, suggesting the participation of Src family kinases in this process. Overall, these results suggest that caveolin-1 plays multiple roles in the VEGF-induced signaling cascade. PMID- 12529450 TI - Active tool use with the contralesional hand can reduce cross-modal extinction of touch on that hand. AB - After a unilateral brain lesion, patients may show cross-modal, visual-tactile extinction. Such patients may fail to report tactile stimuli on the contralesional hand when presented together with competing visual stimuli near the ipsilesional hand. In this work we tested the hypothesis that this cross modal extinction may be reduced when a patient has used a tool with the contralesional hand to reach for objects in the ipsilesional visual field. Consistent with previous work, we hypothesize that active use of a tool may extend cross-modal interactions between visual stimuli at the tip of the tool and tactile stimuli on the hand wielding the tool. In the new situation of a tool connecting the contralesional hand with ipsilesional visual space, competition between stimuli on these opposite sides may be reduced, so that extinction decreases. We studied patient BV, who showed reliable cross-modal, visual-tactile extinction after right-hemisphere stroke. In two separate sessions we showed that prolonged tool use (10-20 min) with the contralesional hand in ipsilesional space reduced cross-modal extinction for up to 60-90 min post-training. We propose that an actively used tool may be effective in linking cross-modal stimuli presented along its extension. This can then overcome competition between stimuli presented on opposite sides of the body midline, thus modulating extinction. PMID- 12529451 TI - Reorganization of speech production at the motor cortex and cerebellum following capsular infarction: a follow-up functional magnetic resonance imaging study. AB - Based on clinical data, Geschwind assumed left hemisphere dominance of speech production to extend to the cortical representation of articulatory and phonatory functions at the motor cortex. This author suggested, furthermore, that the clinical observation of rapid recovery from articulatory impairments after damage to the left-sided corticobulbar tracts reflects compensatory activation of an alternative pathway involving the contralateral pre-central gyrus and its efferent projections. In order to test this hypothesis, functional magnetic resonance imaging (fMRI) was performed 4 and 35 days after stroke in a 38-year old man who had experienced sudden speech deterioration ('dysarthric speech') concomitant with weakness of the right upper limb and the right side of the face. Computerized tomography demonstrated an ischaemic infarction within the left internal capsule. The patient fully recovered from dysarthria within 9 days. Activation of the right hemisphere analogues of Broca and Wernicke areas has been assumed to contribute to recovery from aphasia. As a further aspect of the reorganization of speech function, the present case study demonstrates for the first time by means of fMRI a selective 'shift' of the cortical representation of speech motor control to the right Rolandic cortex and the left cerebellum during restitution of articulation in a case of transient dysarthria following infarction of the left internal capsule. PMID- 12529452 TI - The impact of developmental visuospatial learning difficulties on British Sign Language. AB - There has been substantial research interest in recent years in the relationship between the development of language and cognition, especially where dissociations can be seen between them. Williams syndrome, a rare congenital disorder characterized by a fractionation of higher cortical functions, with relatively preserved language but marked difficulties with visuospatial constructive cognition, has been extensively studied. The case of Heather, who is remarkably similar to the characteristic phenotype of Williams syndrome in physical appearance and cognitive abilities, but who is also congenitally deaf and a user of British Sign Language, provides the first opportunity to explore the consequences of specific visuospatial learning difficulties on the linguistic system when the language used is visuospatial. Heather shows a pattern of impaired drawing ability and visual form discrimination, but preserved ability to discriminate faces. She has a large vocabulary in British Sign Language, and overall presents a picture of relative competence in British Sign Language grammar. However, she shows specific deficits in those areas of British Sign Language which directly rely on spatial representations for linguistic purposes. A number of theories as to the nature of her impairments and those found in Williams syndrome are discussed, using models of the relationship between language and visuospatial cognition based on data from this unique case. PMID- 12529453 TI - Contribution of the left dorsomedial thalamus to recognition memory: a neuropsychological case study. AB - This study reports a patient with a unilateral left thalamic lesion which was centred on the dorsomedial thalamic nucleus. Cognitive neuropsychological assessment revealed a severe impairment in verbal memory and symptoms of executive dysfunction, in the presence of relatively intact visual and facial recognition, working memory, praxis, language and IQ. Verbal and visual recognition memory were investigated using the remember-know paradigm. The results indicated a profound impairment in recollection-driven verbal recognition memory. These results are discussed in the context of the role of the dorsomedial thalamic nucleus in recognition memory, and functional models of memory. PMID- 12529454 TI - Striking a chord with amnesic patients: evidence that song facilitates memory. AB - There have been several reports in the literature, mainly anecdotal, of amnesic patients demonstrating an ability to learn and remember song. Yet there has been little systematic investigation of this phenomenon. In this paper we report findings from three experiments investigating memory for song in two amnesic patients, Joan and Nellie. In the first study we explored recognition memory for various elements of song (i.e. lyrics, melody, lyrics and melody combined) and found that amnesic patients were better than chance at distinguishing studied from novel materials when given explicit instructions. In experiment 2, the ability to generate lyrics and melody was explored in one patient. Although this patient could only generate melodies, her performance in this modality was equivalent to that of matched controls. Memory for the semantic content of song was investigated in a final study using explicit testing conditions. The results showed that both patients were significantly better than chance at identifying correct responses, indicating that they acquired and retained knowledge of the general content of studied songs. These findings not only highlight the special nature of song in facilitating memory in amnesic patients, but given the level of accuracy in memory performance observed, suggest an alternative direction in rehabilitation. PMID- 12529455 TI - A longitudinal study of category-specific agnosia. AB - We report a 12-year longitudinal case study on a 60-year-old male patient (DW) with category-specific agnosia. The extent to which DW's impairment has changed over time was evaluated using identical tests at time 1 (1988) and time 2 (2000). In particular, we assessed his ability to identify pictures and real objects, to draw from memory, and to access stored semantic information about living and non living things. The principal findings were: (i). DW was significantly better at identifying real objects in comparison with line drawings. (ii). DW presented with a category-specific impairment for living things that remained consistent over the 12-year period. (iii). He significantly improved in his ability to identify real non-living objects over the 12-year period but real living objects remained at floor. (iv). His ability to access stored visual knowledge declined over time. On the basis of these data, we suggest that visual perception is required to maintain intact visual memories over a period of time. We also suggest that integrative visual agnosia co-occurs with a category-specific impairment for living things because the recognition of these items requires more global processing than for non-living things. In addition, we suggest that degradation to stored visual knowledge can cause category-specific naming impairments for living compared with non-living things because naming living things requires access to more detailed visual knowledge. PMID- 12529456 TI - A new sign of callosal disconnection syndrome: agonistic dyspraxia. A case study. AB - We report a patient with callosal haemorrhage and no extracallosal involvement who developed a unique form of intermanual conflict. In the acute phase the patient showed a mild speech disturbance and right hemiparesis, and in her right hand, a grasp reflex and compulsive manipulation of tools, all attributable to transient frontal involvement. In the chronic phase there was intermanual conflict occasionally associated with the sensation of a second left hand. The patient also presented a sign consisting of compulsive, automatic execution of orders by one hand (the left or the right) when the patient was specifically asked to perform the movement with the other hand (the right or the left, respectively). There was no left-right confusion in this patient. We call this condition agonistic dyspraxia. In contrast with diagonistic dyspraxia, this consists of the agonistic behaviour of the other hand under conditions in which the hand that has been instructed to respond cannot execute the request. PMID- 12529457 TI - Ensuring safe and effective medical devices. PMID- 12529458 TI - Risks and benefits of gene therapy. PMID- 12529459 TI - Imaging studies after a first febrile urinary tract infection in young children. AB - BACKGROUND: Guidelines from the American Academy of Pediatrics recommend obtaining a voiding cystourethrogram and a renal ultrasonogram for young children after a first urinary tract infection; renal scanning with technetium-99m-labeled dimercaptosuccinic acid has also been endorsed by other authorities. We investigated whether imaging studies altered management or improved outcomes in young children with a first febrile urinary tract infection. METHODS: In a prospective trial involving 309 children (1 to 24 months old), an ultrasonogram and an initial renal scan were obtained within 72 hours after diagnosis, contrast voiding cystourethrography was performed one month later, and renal scanning was repeated six months later. RESULTS: The ultrasonographic results were normal in 88 percent of the children (272 of 309); the identified abnormalities did not modify management. Acute pyelonephritis was diagnosed in 61 percent of the children (190 of 309). Thirty-nine percent of the children who underwent cystourethrography (117 of 302) had vesicoureteral reflux; 96 percent of these children (112 of 117) had grade I, II, or III vesicoureteral reflux. Repeated scans were obtained for 89 percent of the children (275 of 309); renal scarring was noted in 9.5 percent of these children (26 of 275). CONCLUSIONS: An ultrasonogram performed at the time of acute illness is of limited value. A voiding cystourethrogram for the identification of reflux is useful only if antimicrobial prophylaxis is effective in reducing reinfections and renal scarring. Renal scans obtained at presentation identify children with acute pyelonephritis, and scans obtained six months later identify those with renal scarring. The routine performance of urinalysis, urine culture, or both during subsequent febrile illnesses in all children with a previous febrile urinary tract infection will probably obviate the need to obtain either early or late scans. PMID- 12529460 TI - Intratumoral T cells, recurrence, and survival in epithelial ovarian cancer. AB - BACKGROUND: Although tumor-infiltrating T cells have been documented in ovarian carcinoma, a clear association with clinical outcome has not been established. METHODS: We performed immunohistochemical analysis of 186 frozen specimens from advanced-stage ovarian carcinomas to assess the distribution of tumor infiltrating T cells and conducted outcome analyses. Molecular analyses were performed in some tumors by real-time polymerase chain reaction. RESULTS: CD3+ tumor-infiltrating T cells were detected within tumor-cell islets (intratumoral T cells) in 102 of the 186 tumors (54.8 percent); they were undetectable in 72 tumors (38.7 percent); the remaining 12 tumors (6.5 percent) could not be evaluated. There were significant differences in the distributions of progression free survival and overall survival according to the presence or absence of intratumoral T cells (P<0.001 for both comparisons). The five-year overall survival rate was 38.0 percent among patients whose tumors contained T cells and 4.5 percent among patients whose tumors contained no T cells in islets. Significant differences in the distributions of progression-free survival and overall survival according to the presence or absence of intratumoral T cells (P<0.001 for both comparisons) were also seen among 74 patients with a complete clinical response after debulking and platinum-based chemotherapy: the five-year overall survival rate was 73.9 percent among patients whose tumors contained T cells and 11.9 percent among patients whose tumors contained no T cells in islets. The presence of intratumoral T cells independently correlated with delayed recurrence or delayed death in multivariate analysis and was associated with increased expression of interferon-gamma, interleukin-2, and lymphocyte attracting chemokines within the tumor. The absence of intratumoral T cells was associated with increased levels of vascular endothelial growth factor. CONCLUSIONS: The presence of intratumoral T cells correlates with improved clinical outcome in advanced ovarian carcinoma. PMID- 12529461 TI - Pseudomonas aeruginosa and Serratia marcescens contamination associated with a manufacturing defect in bronchoscopes. AB - BACKGROUND: Several outbreaks and pseudo-outbreaks of Pseudomonas aeruginosa and Serratia marcescens infections associated with bronchoscopy have been reported. We conducted an investigation of P. aeruginosa and S. marcescens isolates related to bronchoscopy at a community hospital. METHODS: We reviewed the records of all bronchoscopic procedures at the community hospital from July to October 2001. Environmental samples were obtained. Pulsed-field gel electrophoresis (PFGE) was performed on isolates of P. aeruginosa. RESULTS: From July 1 to October 31, 2001, 66 bronchoscopic procedures were performed in 60 patients, and 43 specimens were obtained for bacterial culture; 20 of the specimens (47 percent) were positive for P. aeruginosa. Six (30 percent) of the specimens that were positive for P. aeruginosa also yielded S. marcescens. All 20 P. aeruginosa isolates were associated with procedures performed with three of four new bronchoscopes from the same manufacturer. Contrary to manufacturing specifications, the biopsy-port caps on all four bronchoscopes were easily removable, and P. aeruginosa was cultured from the biopsy ports of the three implicated bronchoscopes. The PFGE patterns of P. aeruginosa isolates from the bronchoscopes, patients, and two environmental samples were indistinguishable. One patient was hospitalized with P. aeruginosa pneumonia 11 days after bronchoscopy. The manufacturer reported a design change instituted in 1997, and production problems may have resulted in the distribution of bronchoscopes that did not meet specifications. CONCLUSIONS: We documented contamination of bronchoscopes with P. aeruginosa and S. marcescens and possible infection of patients at a community hospital as a result of the inadequate disinfection of bronchoscopes because of a manufacturing defect. PMID- 12529462 TI - An outbreak of Pseudomonas aeruginosa infections associated with flexible bronchoscopes. AB - BACKGROUND: Endoscopes, including bronchoscopes, are the medical devices most frequently associated with outbreaks of nosocomial infections. We investigated an outbreak of Pseudomonas aeruginosa infections after bronchoscopic procedures. METHODS: Microbiologic results were reviewed to determine the rates of recovery of P. aeruginosa from bronchoalveolar-lavage specimens. Environmental samples from endoscopes and the endoscopy suite were cultured. Medical records were reviewed to identify infections in the 14 days after a bronchoscopy. RESULTS: The rate of recovery of P. aeruginosa from bronchoalveolar-lavage specimens obtained with use of endoscopy-suite bronchoscopes increased from 10.4 percent at base line to 31.0 percent during the outbreak (relative risk, 2.97; 95 percent confidence interval, 2.28 to 3.90). Cultures of samples from three bronchoscopes grew P. aeruginosa, whereas cultures of samples from the environment, instrument cleaning machines, and gastrointestinal endoscopes did not. The three bronchoscopes had been part of a nationwide recall. A total of 414 patients underwent bronchoscopy during the outbreak, and there were 48 respiratory tract and bloodstream infections among 39 of these patients (9.4 percent). In 32 infections (66.7 percent), P. aeruginosa was confirmed as a potentially causative organism. Exposure to a potentially contaminated bronchoscope may have had a role in the death of three patients. The rate of recovery of P. aeruginosa returned to base line after the instruments were removed from service. CONCLUSIONS: This large outbreak of P. aeruginosa infections related to bronchoscopy was apparently caused by a loose biopsy-port cap in the bronchoscopes. Instrument safety and surveillance methods for bronchoscopy must be improved, and better recall procedures are needed for medical devices. PMID- 12529463 TI - Images in clinical medicine. Retained surgical instrument. PMID- 12529465 TI - Clinical practice. Suspected appendicitis. PMID- 12529464 TI - Risk factors for retained instruments and sponges after surgery. AB - BACKGROUND: Risk factors for medical errors remain poorly understood. We performed a case-control study of retained foreign bodies in surgical patients in order to identify risk factors for this type of error. METHODS: We reviewed the medical records associated with all claims or incident reports of a retained surgical sponge or instrument filed between 1985 and 2001 with a large malpractice insurer representing one third of the physicians in Massachusetts. For each case, we identified an average of four randomly selected controls who underwent the same type of operation during the same six-month period. RESULTS: Our study included 54 patients with a total of 61 retained foreign bodies (of which 69 percent were sponges and 31 percent instruments) and 235 control patients. Thirty-seven of the patients with retained foreign bodies (69 percent) required reoperation, and one died. Patients with retained foreign bodies were more likely than controls to have had emergency surgery (33 percent vs. 7 percent, P<0.001) or an unexpected change in surgical procedure (34 percent vs. 9 percent, P<0.001). Patients with retained foreign bodies also had a higher mean body-mass index and were less likely to have had counts of sponges and instruments performed. In multivariate analysis, factors associated with a significantly increased risk of retention of a foreign body were emergency surgery (risk ratio, 8.8 [95 percent confidence interval, 2.4 to 31.9]), unplanned change in the operation (risk ratio, 4.1 [95 percent confidence interval, 1.4 to 12.4]), and body-mass index (risk ratio for each one-unit increment, 1.1 [95 percent confidence interval, 1.0 to 1.2]). CONCLUSIONS: The risk of retention of a foreign body after surgery significantly increases in emergencies, with unplanned changes in procedure, and with higher body-mass index. Case--control analysis of medical-malpractice claims may identify and quantify risk factors for specific types of errors. PMID- 12529466 TI - Case record of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 2-2003. A 60-year-old man with mild congestive heart failure of uncertain cause. PMID- 12529467 TI - Imaging studies for childhood urinary infections. PMID- 12529468 TI - Immunosurveillance against cancer and immunotherapy--synergy or antagonism? PMID- 12529469 TI - A serious adverse event after successful gene therapy for X-linked severe combined immunodeficiency. PMID- 12529470 TI - Warfarin, aspirin, or both after myocardial infarction. PMID- 12529471 TI - Botulinum toxin for spasticity after stroke. PMID- 12529472 TI - Peginterferon alfa-2a plus ribavirin for chronic hepatitis C. PMID- 12529473 TI - Increase in nocturnal blood pressure and progression to microalbuminuria in diabetes. PMID- 12529474 TI - Nephrolithiasis, osteoporosis, and mutations in the type 2a sodium-phosphate cotransporter. PMID- 12529475 TI - Insulin-injection-site reactions associated with type I latex allergy. PMID- 12529476 TI - In for the long run: focus on "Lifelong voluntary exercise in the mouse prevents age-related alterations in gene expression in the heart". PMID- 12529478 TI - Measuring activity of the autonomic nervous system in humans. PMID- 12529480 TI - Ethnicity, insurance type, and follow-up in a pediatric weight management program. AB - OBJECTIVE: To assess the characteristics of children and adolescents who drop out of a clinical weight management program. RESEARCH METHODS AND PROCEDURES: A retrospective survey of children and adolescents attending a pediatric weight management program [n = 518; mean +/- SEM: age 10.4 +/- 0.2 years, BMI z-score 4.9 +/- 0.1, 62% female, 41% African American and 57% white] was undertaken. Characteristics of children (age, relative weight, gender, and ethnicity) who dropped out of the program were compared with characteristics of those who continued beyond the initial assessment as a function of type of insurance. RESULTS: The children who dropped out of the program after the initial assessment differed by ethnicity from those who continued in the program: 63% of white children returned for more than one visit, whereas only 35% of the African American children continued in the program. Indemnity insurance coverage was associated with more clinic visits, as compared with managed care insurance. The association between insurance type and clinic visit number was of borderline significance (p = 0.06), when ethnic group was added to this analysis. DISCUSSION: Ethnicity is associated with attendance in a Pediatric Weight Management Program. Given the epidemic of obesity in children and adolescents, issues related to program compliance must be assessed to improve overall outcome. This is especially important given the high rate of obesity among African Americans and the low rate of ongoing attendance observed among African Americans in the Weight Management Program. PMID- 12529479 TI - Adipose tissue quantification by imaging methods: a proposed classification. AB - Recent advances in imaging techniques and understanding of differences in the molecular biology of adipose tissue has rendered classical anatomy obsolete, requiring a new classification of the topography of adipose tissue. Adipose tissue is one of the largest body compartments, yet a classification that defines specific adipose tissue depots based on their anatomic location and related functions is lacking. The absence of an accepted taxonomy poses problems for investigators studying adipose tissue topography and its functional correlates. The aim of this review was to critically examine the literature on imaging of whole body and regional adipose tissue and to create the first systematic classification of adipose tissue topography. Adipose tissue terminology was examined in over 100 original publications. Our analysis revealed inconsistencies in the use of specific definitions, especially for the compartment termed "visceral" adipose tissue. This analysis leads us to propose an updated classification of total body and regional adipose tissue, providing a well defined basis for correlating imaging studies of specific adipose tissue depots with molecular processes. PMID- 12529481 TI - Absence of hypersensitivity to glucocorticoids in antiretroviral-associated lipodystrophy. AB - OBJECTIVE: The lipodystrophy syndrome, which is associated with the use of antiretroviral drugs in some human immunodeficiency virus (HIV)-infected individuals, bears a striking similarity to the fat redistribution observed in Cushing's disease. Although urinary free cortisol excretion and glucocorticoid receptor binding affinity are not elevated in subjects with lipodystrophy, glucocorticoid action at the cellular level has not been examined in affected individuals. The objective of this study was to determine whether tissue sensitivity to glucocorticoids is increased in subjects with lipodystrophy taking protease inhibitors. RESEARCH METHODS AND PROCEDURES: Subjects included 11 HIV infected men on protease inhibitor therapy with lipodystrophy and 10 control HIV infected men not on protease inhibitor therapy and without lipodystrophy. Trunk to extremity fat ratio was measured by DXA. Dexamethasone suppression of peripheral blood mononuclear cell proliferation was measured as an index of tissue sensitivity to glucocorticoid action. RESULTS: Compared with the control group, subjects with lipodystrophy had a significant elevation of the trunk to extremity fat ratio [median (interquartile range): 2.9 (1.3) vs. 1.6 (1.2); p < 0.05]. The concentration of dexamethasone resulting in 50% maximal suppression of proliferation was 11.7 nM (9.3 nM) in subjects with lipodystrophy and 19.6 nM (9.7 nM) in control subjects (p = not significant), and the percentage minimal proliferation was 4% (12%) and 17% (18%) in the two groups, respectively (p = not significant). DISCUSSION: Despite the Cushingoid appearance of affected individuals, these data suggest that body fat redistribution in antiretroviral associated lipodystrophy does not arise through an increase in postreceptor glucocorticoid signaling. PMID- 12529482 TI - Autonomic nervous system activity and the state and development of obesity in Japanese school children. AB - OBJECTIVE: The autonomic nervous system (ANS) plays an important role in regulating energy expenditure and body fat content; however, the extent to which the ANS contributes to pediatric obesity remains inconclusive. The aim of this study was to evaluate whether sympathetic and/or the parasympathetic nerve activities were altered in an obese pediatric population. We further examined a physiological association between the duration of obesity and the sympatho-vagal activities to scrutinize the nature of ANS alteration as a possible etiologic factor of childhood obesity. RESEARCH METHODS AND PROCEDURES: Forty-two obese and 42 non-obese healthy sedentary school children were carefully selected from 1080 participants initially recruited to this study. The two groups were matched in age, gender, and height. The clinical records of physical characteristics and development of the obese children were retrospectively reviewed to investigate the onset and progression of obesity. The ANS activities were assessed during a resting condition by means of heart rate variability power spectral analysis, which enables us to identify separate frequency components, i.e., total power (TP), low-frequency (LF) power, and high-frequency (HF) power. The spectral powers were then logarithmically transformed for statistical testing. RESULTS: The obese children demonstrated a significantly lower TP (6.77 +/- 0.12 vs. 7.11 +/- 0.04 ln ms(2), p < 0.05), LF power (6.16 +/- 0.12 vs. 6.42 +/- 0.05 ln ms(2), p < 0.05), and HF power (5.84 +/- 0.15 vs. 6.34 +/- 0.07 ln ms(2), p < 0.01) compared with the non-obese children. A partial correlation analysis revealed that the LF and HF powers among 42 obese children were negatively associated with the duration of obesity independent of age (LF: partial r = -0.55, p < 0.001; HF: partial r = -0.40, p < 0.01). The obese children were further subdivided into two groups based on the length of their obesity. All three spectral powers were significantly reduced in the obese group with obesity of >3 years (n = 18) compared to the group with obesity of <3 years. DISCUSSION: Our data indicate that obese children possess reduced sympathetic as well as parasympathetic nerve activities. Such autonomic depression, which is associated with the duration of obesity, could be a physiological factor promoting the state and development of obesity. These findings further imply that preventing and treating obesity beginning in the childhood years could be an urgent and crucial pediatric public health issue. PMID- 12529483 TI - Measurement of human daily physical activity. AB - OBJECTIVES: To validate a new device, Intelligent Device for Energy Expenditure and Activity (IDEEA), for the measurement of duration, frequency, and intensity of various types of human physical activity (PA). RESEARCH METHODS AND PROCEDURES: The ability of IDEEA to identify and quantify 32 types of PA, including the most common daily exercise and nonexercise PA, was tested in 76 subjects: Subjects included males (N = 33) and females (N = 43) ranging in age from 13 to 72 years with a mean body mass index (BMI) of 24.7 kg/m(2) (range: 18.4 to 41.0) [43 females: 13 to 72 years old and BMI 18.4 to approximately 41.0 kg/m(2) (mean = 24.7 kg/m(2)); 33 males: 15 to approximately 72 years old and BMI 21.0 to approximately 38.4 kg/m(2) (mean = 25.9 kg/m(2))]. Postures, limb movements, and jumping were tested using a timed protocol of specific activities. Walking and running were tested using a 60-meter track, on which subjects walked and ran at 6 self-selected speeds. Stair climbing and descending were tested by timing subjects who climbed and descended a flight of stairs at two different speeds. RESULTS: Correct identification rates averaged 98.9% for posture and limb movement type and 98.5% for gait type. Pooled correlation between predicted and actual speeds of walking and running was high (r = 0.986, p 2000) of baboons housed at the Southwest Foundation for Biomedical Research in San Antonio, Texas. The long-term goal of this project is to identify genes influencing adiposity-related phenotypes and to test hypotheses regarding their pleiotropic effects on other phenotypes related to increased risk for a variety of common diseases (e.g., coronary heart disease and type 2 diabetes). RESULTS: To date we have obtained various adipose-specific endocrine measures, adipose tissue biopsies, and estimates of body composition on a substantial portion of our pedigreed colony. The pattern of adipose tissue accumulation follows closely that seen in humans, and we have detected significant additive genetic heritabilities for these obesity-related phenotypes. DISCUSSION: Given the physiological and developmental similarities between humans and baboons, along with the ability to collect data under well-controlled situations and the extensive pedigree data available in our colony, the baboon offers an extremely valuable nonhuman primate model for the study of obesity and its comorbidities. PMID- 12529487 TI - Human mesenchymal stem cells as an in vitro model for human adipogenesis. AB - OBJECTIVE: To validate the human mesenchymal stem cells (hMSCs) as a new in vitro model for the study of human adipogenesis, to develop the optimal protocol for the differentiation of hMSCs into adipocytes, and to describe effect of mitogen activated protein kinase on hMSC differentiation into adipocytes. RESEARCH METHODS AND PROCEDURES: hMSCs, obtained commercially, were differentiated by exposure to insulin, dexamethasone, indomethacin, and 3-isobutyl-1-methylxanthine three times for 3 days each. Various differentiation conditions were examined to optimize differentiation as measured by Oil Red O staining. The gene expression during adipogenic conversion was assessed by reverse-transcription polymerase chain reaction, real-time reverse-transcription polymerase chain reaction, and Western blotting. RESULTS: hMSCs differentiated into adipocytes to a different extent depending on the experimental conditions. We have found that differentiation medium based on medium 199 and containing 170 nM insulin, 0.5 mM 3-isobutyl-1-methylxanthine, 0.2 mM indomethacin, 1 microM dexamethasone, and 5% fetal bovine serum was optimal. However, the replacement of fetal bovine serum with rabbit serum (15%) led to further enhancement of differentiation. Inhibition of mitogen-activated protein kinase activation also facilitated adipogenic conversion of hMSCs. The pattern of genes expressed during hMSC differentiation into adipocytes (adipsin, peroxisome proliferator-activated receptor-gamma, CCAAT/enhancer-binding protein-beta, GLUT4, and leptin) was similar to that observed in other in vitro adipocyte models. DISCUSSION: hMSCs are renewable sources of noncommitted precursors that are able to differentiate into mature adipocytes under the proper hormonal and pharmacological stimuli. Thus, hMSCs represent a new model for the study of human adipogenesis. PMID- 12529489 TI - Weight-loss practices and asthma: findings from the behavioral risk factor surveillance system. AB - OBJECTIVE: To describe weight-control practices and receipt of weight-loss advice among obese people with asthma. RESEARCH METHODS AND PROCEDURES: We analyzed data from the 2000 Behavioral Risk Factor Surveillance System. RESULTS: Among 13953 participants with current asthma, 27.3% had a body mass index of >or=30 kg/m2. Overall, 48.1% of participants with asthma reported trying to lose weight (64.1% among overweight or obese participants and 72.9% among obese participants). Among participants with asthma who were trying to lose or maintain weight, 74.7% reported trying to reduce their energy and/or fat intake, and 57.8% reported using physical activity. Approximately 29.7% were using the recommended combination of energy and/or fat intake reduction and physical activity of >or=150 min/wk. During the 12 months before the interview, 16.2% of overweight and 44.9% of obese participants with asthma reported receiving advice to lose weight. Among obese participants receiving weight-loss advice, 82.9% reported trying to lose weight compared to 63.8% of participants who did not receive such advice. DISCUSSION: Health professionals can play an important role in educating their patients with asthma about the importance of weight control and assisting their overweight and obese patients in setting appropriate weight goals and helping them achieve those goals. PMID- 12529490 TI - Turnover of acinar and islet cells in the pancreas of monosodium glutamate treated obese mice. AB - OBJECTIVE: Subcutaneous administrations of monosodium glutamate (MSG) to neonatal animals result in obesity and induce the toxicity on the central nervous system, and furthermore, have an effect on entero-pancreatic hormone. The effect of MSG on the cell turnover of organs, especially the pancreas, has received little attention until now. This study was designed to examine the effect of MSG on pancreatic cell turnover by immunohistochemistry and [(3)H]thymidine autoradiography. RESEARCH METHODS AND PROCEDURES: Male JcI-ICR strain mice were SC injected with MSG (2 mg/g body weight daily) for 5 days after birth, received 112 repeated injections of [(3)H]thymidine at 6-hour intervals for 28 days after birth, and then were killed immediately thereafter, or 30, 60, or 120 days after the last injection. Autoradiography was performed on sections immunostained for glucagon, insulin, and somatostatin. RESULTS: After continuous labeling, most pancreatic cells were labeled, and thereafter, labeling of cells decreased in control and MSG-treated mice. The mean grain counts of acinar cells in MSG treated mice decreased more slowly than those in control mice. On the other hand, those of islet cells, including glucagon, insulin, and somatostatin cells, decreased more rapidly in MSG-treated mice than those in control mice. DISCUSSION: Cell turnover of acinar cells was decelerated and that of islet cells including glucagon, insulin, and somatostatin cells was accelerated in MSG treated mice pancreas. MSG-induced hypothalamic lesions exert the contrary influences on the cell turnover of acinar and islet cells. PMID- 12529491 TI - Obesity, fasting plasma insulin, and C-reactive protein levels in healthy children. AB - OBJECTIVE: Obesity is associated with hyperinsulinemia and increased level of C reactive protein in older children and adults, but little is known about these relationships in very young children. We examined these relationships in healthy 2- to 3-year-old children. RESEARCH METHODS AND PROCEDURES: Analyses were performed on data from 491 healthy 2- to 3-year-old Hispanic children enrolled in a dietary study conducted in New York City, 1992 to 1995. RESULTS: Body mass index (BMI), ponderal index, and sum of four skinfolds were highly correlated (r > 0.75) in both boys and girls. Fasting insulin and glucose levels were only modestly correlated (r = 0.37 for boys and r = 0.28 for girls; p < 0.001 for both), but essentially all of the variability in a calculated index of insulin resistance was attributable to variability in fasting insulin level. The correlations of BMI with fasting insulin level were r = 0.16 (p < 0.05) in boys and r = 0.14 (p < 0.05) in girls. In separate multivariate regression analyses adjusting for age and sex, BMI and ponderal index were associated with fasting plasma insulin level (p < 0.001 for both obesity measures). In multivariate regression analyses adjusting simultaneously for age, sex, and either BMI or ponderal index, fasting insulin level, but not these obesity measures, was associated with C-reactive protein level. DISCUSSION: Obesity is associated with higher fasting insulin level, and fasting insulin is associated with C-reactive protein level, in healthy 2- to 3-year-old children. PMID- 12529492 TI - Larger thigh and hip circumferences are associated with better glucose tolerance: the Hoorn study. AB - OBJECTIVE: A higher waist-to-hip ratio, which can be due to a higher waist circumference, a lower hip circumference, or both, is associated with higher glucose levels and incident diabetes. A lower hip circumference could reflect either lower fat mass or lower muscle mass. Muscle mass might be better reflected by thigh circumference. The aim of this study was to investigate the contributions of thigh and hip circumferences, independent of waist circumference, to measures of glucose metabolism. RESEARCH METHODS AND PROCEDURES: For this cross-sectional study we used baseline data from the Hoorn Study, a population-based cohort study of glucose tolerance among 2484 men and women aged 50 to 75. Glucose tolerance was assessed by a 75-g oral glucose tolerance test; hemoglobin A(1c) and fasting insulin were also measured. Anthropometric measurements included body mass index (BMI) and waist, hip, and thigh circumferences. RESULTS: Stratified analyses and multiple linear regression showed that after adjustment for age, BMI, and waist circumference, thigh circumference was negatively associated with markers of glucose metabolism in women, but not in men. Standardized beta values in women were -0.164 for fasting, -0.206 for post-load glucose, -0.190 for hemoglobin A(1c) (all p < 0.001), and 0.065 for natural log insulin levels (p = 0.061). Hip circumference was negatively associated with markers of glucose metabolism in both sexes (standardized betas ranging from -0.093 to -0.296, p < 0.05) except for insulin in men. Waist circumference was positively associated with glucose metabolism. DISCUSSION: Thigh circumference in women and hip circumference in both sexes are negatively associated with markers of glucose metabolism independently of the waist circumference, BMI, and age. Both fat and muscle tissues may contribute to these associations. PMID- 12529493 TI - Lipoprotein oxidation and plasma vitamin E in nondiabetic normotensive obese patients. AB - OBJECTIVE: To correlate the susceptibility of low-(LDL) and very-low-density lipoprotein to oxidation in vitro and the concentrations of serum antibodies against malondialdehyde-modified LDL and plasma vitamin E with the anthropometric and laboratory characteristics of obesity. RESEARCH METHODS AND PROCEDURES: A total of 75 nondiabetic, normotensive obese patients were assigned to one of four groups according to their body mass index (BMI): moderately obese (30 50 kg/m(2), n = 15). RESULTS: The oxidation lag time for LDL from patients with a BMI >or=35 kg/m(2) was shorter than that for LDL from non-obese controls (n = 13), whereas very-low-density lipoprotein oxidation lag times were not significantly different. The serum antibodies against modified LDL were similar in all groups, whereas the plasma vitamin E concentrations of obese patients were decreased (p or= 85th percentile of age and sex-specific cutoff points from the 2000 Centers for Disease Control and Prevention/National Center for Health Statistics growth charts). In addition, we used coefficients from our logistic regression models to project the effects on overweight prevalence of equalizing the socioeconomic status (SES) differences between race/ethnic groups. RESULTS: Keeping adolescents in their same environments and changing only family income and parental education had a limited effect on the disparities in overweight prevalence. Ethnicity-SES overweight differences were greater among females than males. Given that overweight prevalence decreased with increasing SES among white females and remained elevated and even increased among higher SES African-American females, African-American/white disparity in overweight prevalence increased at the highest SES. Conversely, disparity was lessened at the highest SES for white, Hispanic, and Asian females. Among males, disparity was lowest at the average SES level. DISCUSSION: One cannot automatically assume that the benefits of increased SES found among white adults will transfer to other gender-age-ethnic groups. Our findings suggest that efforts to reduce overweight disparities between ethnic groups must look beyond income and education and focus on other factors, such as environmental, contextual, biological, and sociocultural factors. PMID- 12529495 TI - Vision and eating behavior in obese subjects. AB - OBJECTIVE: Vision is one of a number of factors influencing the amount of food consumed during a meal. The purpose of this study was to investigate the impact of vision on the microstructure of the eating behavior of obese subjects. RESEARCH METHODS AND PROCEDURES: Eighteen obese subjects with a body mass index (mean +/- SD) of 39.1 +/- 6.3 kg/m(2) twice consumed a standardized test meal in excess, once with and once without a blindfold. The microstructure of the eating behavior was registered by VIKTOR, a computerized eating monitor. Subjective motivation to eat (i.e., desire to eat, hunger, satiety, and prospective consumption) was rated by visual analogue scales (VASs) before, immediately after, and then hourly up to 3 hours after the test meals. RESULTS: The obese subjects ate 24% less food when blindfolded (359 +/- 194 g vs. 472 +/- 179 g; p < 0.01). Despite a smaller amount of food consumed when blindfolded, there were no significant differences with or without the blindfold for any of the VASs measuring subjective motivation to eat after test meals. DISCUSSION: The importance of vision in regulating our eating behavior was demonstrated in this study. The obese subjects ate 24% less food blindfolded without feeling less full. Eating blindfolded could be tested as a didactic tool to make obese subjects aware of what factors affect the termination of eating. PMID- 12529496 TI - Discrimination of health risk by combined body mass index and waist circumference. AB - OBJECTIVE: NIH Clinical Guidelines (1998) recommend the measurement of waist circumference (WC, centimeters) within body mass index (BMI, kilograms per square meter) categories as a screening tool for increased health risk. RESEARCH METHODS AND PROCEDURES: The Canada Heart Health Surveys (1986 through 1992) were used to describe the prevalence of the metabolic syndrome in Canada and to test the use of the NIH guidelines for predicting metabolic risk factors. The sample included 7981 participants ages 20 to 74 years who had complete data for WC, BMI, high density lipoprotein-cholesterol, triglycerides, diabetic status, and systolic and diastolic blood pressures. National Cholesterol Education Program Adult Treatment Panel III risk categories were used to identify the metabolic syndrome and associated risk factors. Logistic regression was used to test the hypothesis that WC improves the prediction of the metabolic syndrome, within overweight (25 to 29.9 kg/m(2)) and obese I (30 to 34.9 kg/m(2)) BMI categories. RESULTS: The prevalence of the metabolic syndrome was 17.0% in men and 13.2% in women. The odds ratios (OR) for the prediction of the metabolic syndrome were elevated in overweight [OR, 1.85; 95% confidence interval (95%CI), 1.02 to 3.35] and obese (OR, 2.35; 95%CI, 1.25 to 4.42) women with a high WC compared with overweight and obese women with a low WC, respectively. On the other hand, WC was not predictive of the metabolic syndrome or component risk factors in men, within BMI categories. DISCUSSION: In women already at increased health risk because of an elevated BMI, the additional measurement of WC may help identify cardiovascular risk. PMID- 12529497 TI - Parental weight status and girls' television viewing, snacking, and body mass indexes. AB - OBJECTIVE: The purpose of this study was to examine whether television viewing (TVV) provides a context for patterns of snacking fostering overweight in young girls from overweight and non-overweight families. RESEARCH METHODS AND PROCEDURES: Participants were 173 non-Hispanic white girls and their parents from central Pennsylvania, assessed longitudinally when girls were 5, 7, and 9 years old. Path analysis was used to test patterns of relationships among girls' TVV, snacking while watching television, snacking frequency, fat intake from energy dense snack food, and girls' increase in body mass index (BMI) from age 5 to 9. RESULTS: In both overweight and non-overweight families, girls who watched more television consumed more snacks in front of the television. In families where neither parent was overweight, television viewing was the only significant predictor of girls' increase in BMI. In families where one or both parents were overweight, girls who watched more television snacked more frequently, and girls who snacked more frequently had higher intakes of fat from energy-dense snacks, which predicted their increase in BMI from age 5 to 9. TVV did not directly predict girls' increase in BMI in girls from overweight families. DISCUSSION: The results of this study support and extend previous findings that have shown that excessive television viewing and snacking patterns are risk factors for the development of overweight in children; however, patterns of relationships may differ based on parental weight status. For overweight families, TVV may provide a context for excessive snack consumption, in addition to inactivity. PMID- 12529498 TI - Fat compartments and apolipoprotein B-100 kinetics in overweight-obese men. AB - OBJECTIVE: To examine the association between the kinetics of very-low-density lipoprotein (VLDL)-apolipoprotein B-100 (apoB) and intraperitoneal, retroperitoneal, subcutaneous abdominal, and total adipose tissue masses (IPATM, RPATM, SAATM, and TATM, respectively) in overweight/obese men. RESEARCH METHODS AND PROCEDURES: Hepatic secretion of VLDL was measured using an intravenous infusion of 1-[(13)C]-leucine in 51 men with a wide range of body mass index (25.1 to 42.2 kg/m(2)). Isotopic enrichment of VLDL-apoB was measured using gas chromatography-mass spectrometry and a multicompartmental model used to estimate VLDL-apoB metabolic parameters. IPATM, RPATM, and SAATM (kilograms) were quantified between T11 and S1 using magnetic resonance imaging; TATM (kilograms) was determined using bioelectrical impedance. Insulin resistance was estimated by homeostasis model assessment (HOMA) score. RESULTS: In stepwise regression, IPATM was the best predictor of hepatic secretion of VLDL-apoB (r = 0.390, p < 0.005) and TATM was the best predictor of VLDL-apoB fractional catabolic rate (r = 0.282, p < 0.05). IPATM remained significantly associated with VLDL-apoB secretion after adjusting for TATM or HOMA score (r = 0.360, p < 0.01 and r = 0.310, p < 0.05, respectively). This association was also independent of age, dietary intake, and body mass index. None of the fat compartments were significantly associated with the fractional catabolic rate of VLDL-apoB after adjusting for HOMA score. DISCUSSION: In overweight/obese men, the quantity of both IPATM and TATM determine the kinetics of VLDL-apoB. The effect of IPATM on VLDL-apoB secretion is independent of both total fat mass and the degree of insulin resistance. PMID- 12529499 TI - Plasma leptin and blood pressure in men: graded association independent of body mass and fat pattern. AB - OBJECTIVE: The role of leptin in the association between body mass, central adiposity, and blood pressure (BP) is controversial. This study evaluated the relationship between leptin and BP in relation to body mass index (BMI) and fat distribution in a large sample of untreated male adults. RESEARCH METHODS AND PROCEDURES: The study population was made up of 457 untreated male employees of the Olivetti factory in Naples. Plasma leptin, complete anthropometry, BP, and relevant biochemical variables were measured. RESULTS: Log-transformed plasma leptin levels were directly associated with BMI (r = 0.661, p < 0.001) and waist circumference (r = 0.630; p < 0.001). Leptin also correlated with systolic (r = 0.258) and diastolic (r = 0.277) BP (p < 0.001). The association between leptin and BP was maintained after accounting for age, BMI (or waist circumference), log insulin, and serum creatinine (p < 0.01); this association was stronger than that with BMI. Logistic regression analysis showed that an increased prevalence of hypertension (BP >or= 140 and/or 90 mm Hg) was associated with high plasma leptin levels when controlling for age and waist circumference (odds ratio, 1.99; 95%CI, 1.06 to 3.72) or for age and BMI (odds ratio, 1.92; 95%CI, 1.02 to 3.61). DISCUSSION: A graded positive relationship between plasma leptin levels and BP was observed in this sample of untreated male adults. This association was independent of age, BMI, abdominal adiposity, and fasting plasma insulin. Moreover, elevated plasma leptin concentrations were associated with greater probability of hypertension, again independently of potential confounders. PMID- 12529500 TI - Optimized expression and specific activity of IL-12 by directed molecular evolution. AB - DNA delivery of IL-12 has shown promise in reducing the toxic side effects associated with administration of recombinant human (h)IL-12 protein while maintaining the ability to inhibit tumor growth and abolish tumor metastases in animal models. We have developed a more potent version of IL-12 by using DNA shuffling and screening to improve its expression in human cells and specific activity on human T cells. The most improved evolved IL-12 (EvIL-12) derived from seven mammalian genes encoding both the p35 and p40 subunits of IL-12 showed a 128-fold improvement in human T cell proliferation compared with native hIL-12 during the initial screening of supernatants from transected cells. When purified hIL-12 and EvIL-12 proteins were compared in vitro in human T cell proliferation and Th1 differentiation assays, it was demonstrated that EvIL-12 exhibited a concomitant 10-fold increase in the specific activity of the protein compared with hIL-12. Furthermore, DNA shuffling improved the level of expression and homogeneity of the heterodimer synthesized by 293 human embryonic kidney cells transfected with EvIL-12 by at least 10-fold. Molecular analysis of the variant revealed strategic placement of amino acid substitutions that potentially may facilitate heterodimer formation and product expression. The enhanced expression and biological activity of EvIL-12 may improve the effectiveness of IL-12 gene based vaccines and therapeutics without the toxic side effects sometimes associated with hIL-12 protein administration. PMID- 12529501 TI - A segmental nearest neighbor normalization and gene identification method gives superior results for DNA-array analysis. AB - An intuitive normalization and gene identification method is proposed. After segmentation of the entire expression range into intensity intervals, the mean and standard deviation of the logarithm of expression ratios are calculated for each interval using the nearest neighbor genes. Genes with high differential expression are excluded from these calculations. For glass arrays, normalization is performed for each interval by using the mean of the logarithm of expression ratios in the interval. For nylonplastic membranes, the average of the means of the logarithm of ratios across the intervals of higher intensities is used for normalization. Compared with other normalization methods, this method delivered the smallest normalization errors for 42 nylonplastic arrays used to analyze cultured T cells and 22 Clostridium acetobutylicum glass arrays. For identifying differentially expressed genes, upper and lower boundaries are constructed for each interval by using the standard deviation of the expression ratio logarithms. When a C. acetobutylicum pSOL1 megaplasmid-deficient strain M5 was used, this method identified more "down-regulated" pSOL1 genes with fewer misidentifications in a comparative array analysis of M5 versus the parent strain. A comparison of quantitative RT-PCR results with different gene identification methods indicates that the proposed method is superior to other methods. PMID- 12529502 TI - No hypothermic response to serotonin in 5-HT7 receptor knockout mice. AB - With data from recently available selective antagonists for the 5-HT(7) receptor, it has been hypothesized that 5-hydroxytryptamine (5-HT)-induced hypothermia is mediated by the 5-HT(7) receptor, an effect previously attributed to other receptor subtypes. It has been established that the biologically active lipid oleamide allosterically interacts with the 5-HT(7) receptor to regulate its transmission. The most well characterized effects of oleamide administration are induction of sleep and hypothermia. Here, we demonstrate, by using mice lacking the 5-HT(7) receptor, that 5-HT-induced hypothermia is mediated by the 5-HT(7) receptor. Both 5-HT and 5-carboxamidotryptamine, a 5-HT(1) and 5-HT(7) receptor agonist, in physiological doses fail to induce hypothermia in 5-HT(7) knockout mice. In contrast, oleamide was equally effective in inducing hypothermia in mice lacking the 5-HT(7) receptors as in wild-type mice. When administered together, 5 HT and oleamide showed additive or greater than additive effects in reducing body temperature. Taken together, the results show that 5-HT-induced hypothermia is mediated by the 5-HT(7) receptor, and that oleamide may act through an independent mechanism as well as at an allosteric 5-HT(7) receptor site to regulate body temperature. PMID- 12529503 TI - Observation of liquid-liquid phase separation for eye lens gammaS-crystallin. AB - gammaS-crystallin (gammaS) is an important human and bovine eye lens protein involved in maintaining the transparency of the eye. By adding small amounts of polyethylene glycol (PEG) to the binary aqueous bovine gammaS solutions, we have observed liquid-liquid phase separation (LLPS) at -8 degrees C and revealed that, in the binary gammaS-water system, this phase transition would occur at -28 degrees C. We have measured both the effect of PEG concentration on the LLPS temperature and proteinPEG partitioning between the two liquid coexisting phases. We use our measurements of proteinPEG partitioning to determine the nature and the magnitude of the gammaS-PEG interactions and to quantitatively assess the effectiveness of PEG as a crystallizing agent for gammaS. We use our measurements of LLPS temperature as a function of protein and PEG concentration to successfully determine the location of the critical point for the binary gammaS water system. This phase transition cannot be observed in the absence of PEG because it is inaccessible due to the freezing of the system. Our findings indicate that the effective interactions between gammaS molecules in the binary gammaS-water system are attractive. We compare the magnitude of the attraction found for gammaS with the results obtained for the other gamma-crystallins for which the critical temperature is located above the freezing point of the system. This work suggests that PEG can be used to reveal the existence of LLPS for a much wider range of binary protein-water systems than known previously. PMID- 12529504 TI - CTXphi-independent production of the RS1 satellite phage by Vibrio cholerae. AB - The cholera toxin genes of Vibrio cholerae are encoded by the filamentous phage, CTXphi. Chromosomal CTXphi prophage DNA is often found flanked by copies of a related genetic element designated RS1, and RS1 DNA can be packaged into filamentous phage particles (designated RS1phi) by using the CTXphi morphogenesis genes. RS1phi is a satellite phage that further controls expression and dissemination of CTXphi. Here we describe a CTXphi-independent mechanism for production of RS1phi. A nontoxigenic environmental V. cholerae strain (55V71) was identified that supports production of RS1phi. However, newly infected CTX negative strains did not produce RS1phi, indicating that additional 55V71 genes were involved in production of RS1phi. Analysis of nucleic acids from phage preparations of 55V71 revealed a 7.5-kb single-stranded DNA, whose corresponding replicative form was found in plasmid preparations. This DNA likely corresponds to the genome of a new filamentous phage, which we have designated KSF-1phi. The replicative form DNA of KSF-1phi was cloned into pUC18, and the resulting construct pKSF-1.1 supported the production of RS1phi particles by CTX-negative V. cholerae strains. RS1phi particles produced in this way infect recipient V. cholerae strains by a mechanism that is independent of the CTXphi receptor, the toxin-coregulated pilus. Thus, KSF-1phi is capable of facilitating the transfer of the RS1 element to strains that do not express toxin coregulated pilus. Given that RS1phi can enhance coproduction of CTXphi particles, KSF-1phi-mediated dissemination of RS1 may indirectly promote the spread of toxin genes among V. cholerae strains. This study also shows that filamentous phages can package diverse DNA elements and thus may play a role in horizontal transfer of more genes than previously appreciated. PMID- 12529505 TI - Reduction of cholera in Bangladeshi villages by simple filtration. AB - Based on results of ecological studies demonstrating that Vibrio cholerae, the etiological agent of epidemic cholera, is commensal to zooplankton, notably copepods, a simple filtration procedure was developed whereby zooplankton, most phytoplankton, and particulates >20 microm were removed from water before use. Effective deployment of this filtration procedure, from September 1999 through July 2002 in 65 villages of rural Bangladesh, of which the total population for the entire study comprised approximately 133,000 individuals, yielded a 48% reduction in cholera (P < 0.005) compared with the control. PMID- 12529508 TI - Wallenberg Prize acceptance speech. The future of biology: reason for concern? PMID- 12529506 TI - Activation of human eosinophils through leukocyte immunoglobulin-like receptor 7. AB - Eosinophils are implicated prominently in allergic diseases and the host response to parasitic infections. Eosinophils may be activated in vitro by diverse classes of agonists such as immunoglobulins, lipid mediators, and cytokines. The leukocyte Ig-like receptors (LIRs) comprise a family of inhibitory and activating cell-surface receptors. Inhibitory LIRs down-regulate cellular responses through cytoplasmic immunoreceptor tyrosine-based inhibitory motifs. There are limited data on the action of the activating LIRs, which are thought to signal through the Fc receptor gamma chain, which contains an immunoreceptor tyrosine-based activation motif. We now demonstrate the expression of LIR1 (inhibitory), LIR2 (inhibitory), LIR3 (inhibitory), and LIR7 (activating) on eosinophils from 4, 4, 12, and 11, respectively, of 12 healthy donors. Cross-linking of LIR7 with plate bound antibody elicited the dose- and time-dependent release of eosinophil derived neurotoxin and leukotriene C(4). Eosinophils activated with antibodies to LIR7 embedded in gel-phase EliCell preparations showed leukotriene C(4) generation at the nuclear envelope and the release of IL-12 but not IL-4 by vesicular transport. Thus, LIR7 is an activating receptor for eosinophils that elicited the release of cytotoxic granule proteins, de novo lipid mediator generation, and cytokine release through vesicular transport. PMID- 12529507 TI - Identification of a gene causing human cytochrome c oxidase deficiency by integrative genomics. AB - Identifying the genes responsible for human diseases requires combining information about gene position with clues about biological function. The recent availability of whole-genome data sets of RNA and protein expression provides powerful new sources of functional insight. Here we illustrate how such data sets can expedite disease-gene discovery, by using them to identify the gene causing Leigh syndrome, French-Canadian type (LSFC, Online Mendelian Inheritance in Man no. 220111), a human cytochrome c oxidase deficiency that maps to chromosome 2p16 21. Using four public RNA expression data sets, we assigned to all human genes a "score" reflecting their similarity in RNA-expression profiles to known mitochondrial genes. Using a large survey of organellar proteomics, we similarly classified human genes according to the likelihood of their protein product being associated with the mitochondrion. By intersecting this information with the relevant genomic region, we identified a single clear candidate gene, LRPPRC. Resequencing identified two mutations on two independent haplotypes, providing definitive genetic proof that LRPPRC indeed causes LSFC. LRPPRC encodes an mRNA binding protein likely involved with mtDNA transcript processing, suggesting an additional mechanism of mitochondrial pathophysiology. Similar strategies to integrate diverse genomic information can be applied likewise to other disease pathways and will become increasingly powerful with the growing wealth of diverse, functional genomics data. PMID- 12529509 TI - Modulation of guard cell plasma membrane potassium currents by methyl jasmonate. PMID- 12529510 TI - Conservation of the cold shock domain protein family in plants. PMID- 12529511 TI - ARAMEMNON, a novel database for Arabidopsis integral membrane proteins. AB - A specialized database (DB) for Arabidopsis membrane proteins, ARAMEMNON, was designed that facilitates the interpretation of gene and protein sequence data by integrating features that are presently only available from individual sources. Using several publicly available prediction programs, putative integral membrane proteins were identified among the approximately 25,500 proteins in the Arabidopsis genome DBs. By averaging the predictions from seven programs, approximately 6,500 proteins were classified as transmembrane (TM) candidate proteins. Some 1,800 of these contain at least four TM spans and are possibly linked to transport functions. The ARAMEMNON DB enables direct comparison of the predictions of seven different TM span computation programs and the predictions of subcellular localization by eight signal peptide recognition programs. A special function displays the proteins related to the query and dynamically generates a protein family structure. As a first set of proteins from other organisms, all of the approximately 700 putative membrane proteins were extracted from the genome of the cyanobacterium Synechocystis sp. and incorporated in the ARAMEMNON DB. The ARAMEMNON DB is accessible at the URL http://aramemnon.botanik.uni-koeln.de. PMID- 12529512 TI - Identification and characterization of the ARIADNE gene family in Arabidopsis. A group of putative E3 ligases. AB - ARIADNE (ARI) proteins were recently identified in fruitfly (Drosophila melanogaster), mouse, and man because of their specific interaction with the ubiquitin-conjugating (E2) enzymes UbcD10, UbcM4, UbcH7, and UbcH8. They are characterized by specific motifs and protein structures that they share with PARKIN, and there is increasing evidence that ARI/PARKIN proteins function as E2 dependent ubiquitin-protein ligases. On the basis of homology and motif searches, 16 AtARI genes were identified in Arabidopsis. Analysis of the position of exons/introns and their chromosomal localization indicates that the AtARI gene family expanded via larger and smaller genome duplications. We present evidence that retroposition of processed mRNA may have also contributed to enlarging this gene family. Phylogenetic analyses divides the AtARI proteins into three subgroups. Two groups are absent in yeast, invertebrates, and vertebrates and may therefore represent new plant-specific subfamilies. Examination of the predicted protein sequences revealed that the ARI proteins share an additional leucine-rich region at the N terminus that is highly conserved in all phyla analyzed. Furthermore, conserved consensus signals for casein kinase II-dependent phosphorylation and for nuclear localization were identified. The in silico-based analyses were complemented with experimental data to quantify expression levels. Using real-time polymerase chain reaction, we show that the ARI genes are differentially transcribed. AtARI1 is highly expressed in all organs, whereas no transcripts could be detected for AtARI11, AtARI13, and AtARI14. AtARI12 and AtARI16 are expressed in an organ-specific manner in the roots and siliques, respectively. PMID- 12529513 TI - Pit membrane porosity and water stress-induced cavitation in four co-existing dry rainforest tree species. AB - Aspects of xylem anatomy and vulnerability to water stress-induced embolism were examined in stems of two drought-deciduous species, Brachychiton australis (Schott and Endl.) A. Terracc. and Cochlospermum gillivraei Benth., and two evergreen species, Alphitonia excelsa (Fenzal) Benth. and Austromyrtus bidwillii (Benth.) Burret., growing in a seasonally dry rainforest. The deciduous species were more vulnerable to water stress-induced xylem embolism. B. australis and C. gillivraei reached a 50% loss of hydraulic conductivity at -3.17 MPa and -1.44 MPa, respectively; a 50% loss of hydraulic conductivity occurred at -5.56 MPa in A. excelsa and -5.12 MPa in A. bidwillii. To determine whether pit membrane porosity was responsible for greater vulnerability to embolism (air seeding hypothesis), pit membrane structure was examined. Expected pore sizes were calculated from vulnerability curves; however, the predicted inter-specific variation in pore sizes was not detected using scanning electron microscopy (pores were not visible to a resolution of 20 nm). Suspensions of colloidal gold particles were then perfused through branch sections. These experiments indicated that pit membrane pores were between 5 and 20 nm in diameter in all four species. The results may be explained by three possibilities: (a) the pores of the expected size range were not present, (b) larger pores, within the size range to cause air seeding, were present but were rare enough to avoid detection, or (c) pore sizes in the expected range only develop while the membrane is under mechanical stress (during air seeding) due to stretching/flexing. PMID- 12529514 TI - The effects of polyethylene glycol on gene expression of developing white spruce somatic embryos. AB - Somatic embryogenic cultures of white spruce (Picea glauca) represent a valuable system to study molecular mechanisms regulating embryo development because many embryos of defined developmental stages can be generated. The inclusion of polyethylene glycol (PEG) in the maturation medium can improve the number and quality of embryos produced. To learn more about the mechanism of action of PEG, we analyzed transcript profiles of stage-specific embryos matured without (control) or with (PEG treated) PEG. RNA extracted from maturing spruce embryos was analyzed on DNA microarrays containing 2,178 cDNAs from loblolly pine (Pinus taeda). The efficiency of heterologous hybridization between spruce and pine species on microarrays has been documented previously (L. van Zyl, S. von Arnold, P. Bozhkov, Y. Chen, U. Egertsdotter, J. MacKay, R. Sederoff, J. Shen, L. Zelena, D. Clapham [2002] Comp Funct Genomics 3: 306-318). Several pine genes, including the apparent homologs to the Arabidopsis genes ZWILLE, FIDDLEHEAD, FUSCA, and SCARECROW, increased in expression after PEG treatments. These genes are known to be involved in the formation of the embryo body plan and in the control of the shoot and root apical meristems. The increased transcript levels of these genes in immature PEG-treated embryos suggest that PEG may improve the quality of spruce somatic embryos by promoting normal differentiation of the embryonic shoot and root. Changes in the transcript levels of many genes involved in sucrose catabolism and nitrogen assimilation and utilization were also observed between control and PEG-treated embryos. PMID- 12529515 TI - The companion cell-specific Arabidopsis disaccharide carrier AtSUC2 is expressed in nematode-induced syncytia. AB - Cyst nematodes induce a metabolically highly active syncytial cell complex in host roots. The syncytia are symplastically isolated. Because they form a strong sink, assimilates must be imported via the apoplast, thus suggesting that specific membrane-bound sugar transport proteins are expressed and activated. To identify possible candidate genes, transgenic Arabidopsis plants expressing different reporter genes under the control of different promoters from Arabidopsis sugar transporter genes were infected with the beet cyst nematode (Heterodera schachtii). With polymerase chain reaction, 13 additional sugar transporters were tested for their presence in the syncytia through the use of a syncytium-specific cDNA library. Analysis of the infected roots showed that the promoter of the sucrose (Suc) transporter AtSUC2 gene that codes for a companion cell-specific Suc transporter in noninfected plants was found to be expressed in syncytia. Its expression patterns in beta-glucuronidase and green fluorescent protein plants were monitored. Syncytium-specific gene expression was confirmed by reverse transcriptase-polymerase chain reaction. Results support the idea that AtSUC2 mediates the transmembrane transfer of Suc. AtSUC2 is the first disaccharide carrier described to be activated by pathogens. PMID- 12529516 TI - AtSTP6, a new pollen-specific H+-monosaccharide symporter from Arabidopsis. AB - This paper describes the molecular, kinetic, and physiological characterization of AtSTP6, a new member of the Arabidopsis H(+)/monosaccharide transporter family. The AtSTP6 gene (At3g05960) is interrupted by two introns and encodes a protein of 507 amino acids containing 12 putative transmembrane helices. Expression in yeast (Saccharomyces cerevisiae) shows that AtSTP6 is a high affinity (K(m) = 20 microM), broad-spectrum, and uncoupler-sensitive monosaccharide transporter that is targeted to the plasma membrane and that can complement a growth deficiency resulting from the disruption of most yeast hexose transporter genes. Analyses of AtSTP6-promoter::GUS plants and in situ hybridization experiments detected AtSTP6 expression only during the late stages of pollen development. A transposon-tagged Arabidopsis mutant was isolated and homozygous plants were analyzed for potential effects of the Atstp6 mutation on pollen viability, pollen germination, fertilization, and seed production. However, differences between wild-type and mutant plants could not be observed. PMID- 12529517 TI - Regulatory networks in seeds integrating developmental, abscisic acid, sugar, and light signaling. AB - Progression through embryogenesis and the transition to germination is subject to regulation by many transcription factors, including those encoded by the Arabidopsis LEC1 (LEAFY COTYLEDON1), FUS3 (FUSCA3), and abscisic acid-insensitive (ABI) ABI3, ABI4, and ABI5 loci. To determine whether the ABI4, ABI5, LEC1, and FUS3 loci interact or act independently, we analyzed abi fus3 and abi lec1 double mutants. Our results show that both ABI4 and ABI5 interact genetically with both LEC1 and FUS3 in controlling pigment accumulation, suppression of vivipary, germination sensitivity to abscisic acid, gene expression during mid- and late embryogenesis, sugar metabolism, sensitivity to sugar, and etiolated growth. However, the relative strengths of the observed interactions vary among responses and may even be antagonistic. Furthermore, the interactions reveal cryptic effects of individual loci that are not detectable by analyses of single mutants. Despite these strong genetic interactions, but consistent with the disparities in peak expression of these loci, none of the ABI transcription factors appear to interact directly with either FUS3 or LEC1 in a yeast (Saccharomyces cerevisiae) two-hybrid assay system. PMID- 12529518 TI - Elicitor activity of a fungal endopolygalacturonase in tobacco requires a functional catalytic site and cell wall localization. AB - CLPG1, an endopolygalacturonase (endoPG) gene of Colletotrichum lindemuthianum, was transferred to tobacco (Nicotiana tabacum) leaves by using the Agrobacterium tumefaciens transient delivery system. The following four constructs were prepared: CLPG1, with or without its signal peptide (SP; PG1, PG1deltaSP); CLPG1 with the tobacco expansin1 SP instead of its own SP (Exp::PG1deltaSP); and a mutated version of the latter on two amino acids potentially involved in the catalytic site of CLPG1 (D202N/D203N). Chlorotic and necrotic lesions appeared 5 to 7 d postinfiltration, exclusively in response to CLPG1 fused to the expansin SP. The lesions were correlated to the production of an active enzyme. Necrosis inducing activity, as well as endoPG activity, were completely abolished by site directed mutagenesis. Ultrastructural immunocytolocalization experiments indicated that the expansin SP addressed CLPG1 to the cell wall. Staining of parenchyma cells revealed the progressive degradation of pectic material in junction zones and middle lamella as a function of time after infiltration, ultimately leading to cell separation. A 30% decrease in the GalUA content of the cell walls was simultaneously recorded, thereby confirming the hydrolytic effect of CLPG1 on pectic polysaccharides, in planta. The elicitor activity of CLPG1 was further illustrated by the induction of defense responses comprising active oxygen species and beta-1,3-glucanase activity, before leaf necrosis. Altogether, the data demonstrate that an appropriate SP and a functional catalytic site are required for the proper expression and elicitor activity of the fungal endoPG CLPG1 in tobacco. PMID- 12529519 TI - The sucrose transporter StSUT1 localizes to sieve elements in potato tuber phloem and influences tuber physiology and development. AB - The sucrose (Suc) H(+)-cotransporter StSUT1 from potato (Solanum tuberosum), which is essential for long-distance transport of Suc and assumed to play a role in phloem loading in mature leaves, was found to be expressed in sink tubers. To answer the question of whether SUT1 serves a function in phloem unloading in tubers, the promoter was fused to gusA and expression was analyzed in transgenic potato. SUT1 expression was unexpectedly detected not in tuber parenchyma but in the phloem of sink tubers. Immunolocalization demonstrated that StSUT1 protein was present only in sieve elements of sink tubers, cells normally involved in export of Suc from the phloem to supply developing tubers, raising the question of the role of SUT1 in tubers. SUT1 expression was inhibited by antisense in transgenic potato plants using a class I patatin promoter B33, which is primarily expressed in the phloem of developing tubers. Reduced SUT1 expression in tubers did not affect aboveground organs but led to reduced fresh weight accumulation during early stages of tuber development, indicating that in this phase SUT1 plays an important role for sugar transport. Changes in Suc- and starch-modifying enzyme activities and metabolite profiles are consistent with the developmental switch in unloading mechanisms. Altogether, the findings may suggest a role of SUT1 in retrieval of Suc from the apoplasm, thereby regulating the osmotic potential in the extracellular space, or a direct role in phloem unloading acting as a phloem exporter transferring Suc from the sieve elements into the apoplasm. PMID- 12529520 TI - Diversity of the superfamily of phloem lectins (phloem protein 2) in angiosperms. AB - Phloem protein 2 (PP2) is one of the most abundant and enigmatic proteins in the phloem sap. Although thought to be associated with structural P-protein, PP2 is translocated in the assimilate stream where its lectin activity or RNA-binding properties can exert effects over long distances. Analyzing the diversity of these proteins in vascular plants led to the identification of PP2-like genes in species from 17 angiosperm and gymnosperm genera. This wide distribution of PP2 genes in the plant kingdom indicates that they are ancient and common in vascular plants. Their presence in cereals and gymnosperms, both of which lack structural P-protein, also supports a wider role for these proteins. Within this superfamily, PP2 proteins have considerable size polymorphism. This is attributable to variability in the length of the amino terminus that extends from a highly conserved domain. The conserved PP2 domain was identified in the proteins encoded by six genes from several cucurbits, celery (Apium graveolens), and Arabidopsis that are specifically expressed in the sieve element-companion cell complex. The acquisition of additional modular domains in the amino-terminal extensions of other PP2-like proteins could reflect divergence from its phloem function. PMID- 12529521 TI - trans-Resveratrol and grape disease resistance. A dynamical study by high resolution laser-based techniques. AB - Two modern laser-based techniques were synchronously applied to study the dynamics of the trans-resveratrol activity in Botrytis cinerea-infected grapes. Direct analysis of trans-resveratrol in both infected and noninfected grapes (Vitis vinifera, Aledo variety) was performed by using an analytical technique incorporating laser desorption coupled with laser resonant ionization and time-of flight mass spectrometry. On the other hand, one of the most sensitive on-line methods for trace gas detection, laser photoacoustic spectroscopy, was used to investigate the involvement of the plant hormone ethylene (C(2)H(4)) in the B. cinerea grapes interaction and its temporal relationship with the trans resveratrol content upon infection. The trans-resveratrol content and the ethylene released by noninfected grapes showed an opposite behavior. In this case, a high trans-resveratrol content corresponds to a low ethylene emission. For the B. cinerea-infected grapes, ethylene emission rises up after 48 h when the analogous content of trans-resveratrol started to decrease irreversibly. Moreover, the activity of trans-resveratrol as natural pesticide has been investigated by exogenous application on grapes. A short submerge (5 s) of the grapes in 1.6 x 10(-4) M solution of trans-resveratrol delays the increase of C(2)H(4) emission with about 48 h and produces a decrease of the C(2)H(4) concentration and its emission rate. The treatment has positive effects on fruit conservation during storage; it doubled the normal shelf-life of grapes at room temperature, maintaining their post-harvest quality within 10 d. PMID- 12529522 TI - Ethylmethanesulfonate saturation mutagenesis in Arabidopsis to determine frequency of herbicide resistance. AB - Plant resistance to glyphosate has been reported far less frequently than resistance to sulfonylurea and imidazolinone herbicides. However, these studies tend to be anecdotal, without side by side comparisons for a single species or natural isolate. In this study, we tested the frequencies of resistance of three herbicides in a controlled ethylmethanesulfonate (EMS) saturation mutagenesis experiment, allowing a direct comparison of the frequencies at which resistant mutant plants arise. The 100% growth inhibition dose rates of glyphosate, chlorsulfuron (a sulfonylurea herbicide), and imazethapyr (an imidazolinone herbicide) were determined for Arabidopsis. Populations of EMS-mutagenized M(2) seedlings were sprayed with twice the 100% growth inhibition dose of glyphosate, chlorsulfuron, or imazethapyr, and herbicide-resistant mutants were identified. Although there were no glyphosate-resistant mutants among M(2) progeny of 125,000 Columbia and 125,000 Landsberg erecta M(1) lines, chlorsulfuron resistance and imazethapyr resistance each appeared at frequencies of 3.2 x 10(-5). Given the observed frequency of herbicide resistance mutations, we calculate that there are at least 700 mutations in each EMS-mutagenized Arabidopsis line and that fewer than 50,000 M(1) lines are needed to have a 95% chance of finding a mutation in any given G:C base pair in the genome. As part of this study, two previously unreported Arabidopsis mutations conferring resistance to imidazolinone herbicides, csr1-5 (Ala-122-Thr) and csr1-6 (Ala-205-Val), were discovered. Neither of these mutations caused enhanced resistance to chlorsulfuron in Arabidopsis. PMID- 12529523 TI - Extracellular ATP inhibits root gravitropism at concentrations that inhibit polar auxin transport. AB - Raising the level of extracellular ATP to mM concentrations similar to those found inside cells can block gravitropism of Arabidopsis roots. When plants are grown in Murashige and Skoog medium supplied with 1 mM ATP, their roots grow horizontally instead of growing straight down. Medium with 2 mM ATP induces root curling, and 3 mM ATP stimulates lateral root growth. When plants are transferred to medium containing exogenous ATP, the gravity response is reduced or in some cases completely blocked by ATP. Equivalent concentrations of ADP or inorganic phosphate have slight but usually statistically insignificant effects, suggesting the specificity of ATP in these responses. The ATP effects may be attributable to the disturbance of auxin distribution in roots by exogenously applied ATP, because extracellular ATP can alter the pattern of auxin-induced gene expression in DR5-beta-glucuronidase transgenic plants and increase the response sensitivity of plant roots to exogenously added auxin. The presence of extracellular ATP also decreases basipetal auxin transport in a dose-dependent fashion in both maize (Zea mays) and Arabidopsis roots and increases the retention of [(3)H]indole-3 acetic acid in root tips of maize. Taken together, these results suggest that the inhibitory effects of extracellular ATP on auxin distribution may happen at the level of auxin export. The potential role of the trans-plasma membrane ATP gradient in auxin export and plant root gravitropism is discussed. PMID- 12529524 TI - Molecular cloning and biological activity of alpha-, beta-, and gamma megaspermin, three elicitins secreted by Phytophthora megasperma H20. AB - We report on the molecular cloning of the Phytophthora megasperma H20 (PmH20) glycoprotein shown previously as an inducer of the hypersensitive response, of localized acquired resistance and of systemic acquired resistance in tobacco (Nicotiana tabacum), and of the PmH20 alpha- and beta-megaspermin, two elicitins of class I-A and I-B, respectively. The structure of the glycoprotein shows a signal peptide of 20 amino acids followed by the typical elicitin 98-amino acid long domain and a 77-amino acid-long C-terminal domain carrying an O-glycosylated moiety. The molecular mass deduced from the translated cDNA sequence is 14,920 and 18,676 D as determined by mass spectrometry. This structure together with multiple sequence alignments and phylogenetic analyses indicate that the glycoprotein belongs to class III elicitins. It is the first class III elicitin protein characterized, which we named gamma-megaspermin. We compared the biological activity of the three PmH20 elicitins when applied to tobacco cv Samsun NN plants. Although alpha- and gamma-megaspermin were similarly active, beta-megaspermin was the most active in inducing the hypersensitive response and localized acquired resistance, which was assessed by measuring the levels of acidic and basic pathogenesis-related proteins and of the antioxidant phytoalexin scopoletin. The three elicitins induced similar levels of systemic acquired resistance measured as the expression of acidic PR proteins and is increased resistance to challenge tobacco mosaic virus infection. PMID- 12529525 TI - The Arabidopsis AtIPT8/PGA22 gene encodes an isopentenyl transferase that is involved in de novo cytokinin biosynthesis. AB - Cytokinin plays a critical role in plant growth and development by stimulating cell division and cell differentiation. Despite many years' research efforts, our current understanding of this hormone is still limited regarding both its biosynthesis and signaling. To genetically dissect the cytokinin pathway, we have used a functional screen to identify Arabidopsis gain-of-function mutations that enable shoot formation in the absence of exogenous cytokinins. By using a chemical-inducible activation tagging system, we have identified over 40 putative mutants, designated as pga (plant growth activators), which presumably were affected in key components of cytokinin biosynthesis and signaling pathway. Here, we report a detailed characterization of pga22, a representative mutant from this collection. A gain-of-function mutation in the PGA22 locus resulted in typical cytokinin responses. Molecular and genetic analyses indicated that PGA22 encodes an isopentenyl transferase (IPT) previously identified as AtIPT8. Plants of the pga22 mutant accumulated at remarkably higher levels of isopentenyladenosine-5' monophosphate and isopentenyladenosine when analyzed by mass spectrometry, suggesting that AtIPT8/PGA22 is a functional IPT that may direct the biosynthesis of cytokinins in planta via an isopentenyladenosine-5'-monophosphate-dependent pathway. PMID- 12529526 TI - Spatial and temporal effects of free-air CO2 enrichment (POPFACE) on leaf growth, cell expansion, and cell production in a closed canopy of poplar. AB - Leaf expansion in the fast-growing tree, Populus x euramericana was stimulated by elevated [CO(2)] in a closed-canopy forest plantation, exposed using a free air CO(2) enrichment technique enabling long-term experimentation in field conditions. The effects of elevated [CO(2)] over time were characterized and related to the leaf plastochron index (LPI), and showed that leaf expansion was stimulated at very early (LPI, 0-3) and late (LPI, 6-8) stages in development. Early and late effects of elevated [CO(2)] were largely the result of increased cell expansion and increased cell production, respectively. Spatial effects of elevated [CO(2)] were also marked and increased final leaf size resulted from an effect on leaf area, but not leaf length, demonstrating changed leaf shape in response to [CO(2)]. Leaves exhibited a basipetal gradient of leaf development, investigated by defining seven interveinal areas, with growth ceasing first at the leaf tip. Interestingly, and in contrast to other reports, no spatial differences in epidermal cell size were apparent across the lamina, whereas a clear basipetal gradient in cell production rate was found. These data suggest that the rate and timing of cell production was more important in determining leaf shape, given the constant cell size across the leaf lamina. The effect of elevated [CO(2)] imposed on this developmental gradient suggested that leaf cell production continued longer in elevated [CO(2)] and that basal increases in cell production rate were also more important than altered cell expansion for increased final leaf size and altered leaf shape in elevated [CO(2)]. PMID- 12529527 TI - Regulation of early tomato fruit development by the diageotropica gene. AB - The vegetative phenotype of the auxin-resistant diageotropica (dgt) mutant of tomato (Lycopersicon esculentum Mill.) includes reduced gravitropic response, shortened internodes, lack of lateral roots, and retarded vascular development. Here, we report that early fruit development is also dramatically altered by the single-gene dgt lesion. Fruit weight, fruit set, and numbers of locules and seeds are reduced in dgt. In addition, time to flowering and time from anthesis to the onset of fruit ripening are increased by the dgt lesion, whereas ripening is normal. The dgt mutation appears to affect only the early stages of fruit development, irrespective of allele or genetic background. Expression of members of the LeACS (1-aminocyclopropane-1-carboxylic acid synthase, a key regulatory enzyme of ethylene biosynthesis) and LeIAA (Aux/IAA, auxin-responsive) gene families were quantified via real-time reverse transcriptase-polymerase chain reaction in both dgt and wild-type fruits, providing the first analysis of Aux/IAA gene expression in fruit. The dgt lesion affects the expression of only certain members of both the LeACS and LeIAA multigene families. Different subsets of LeIAA gene family members are affected by the dgt mutation in fruits and hypocotyls, indicating that the DGT gene product functions in a developmentally specific manner. The differential expression of subsets of LeIAA and LeACS gene family members as well as the alterations in dgt fruit morphology and growth suggest that the early stages of fruit development in tomato are regulated, at least in part, by auxin- and ethylene-mediated gene expression. PMID- 12529528 TI - Proteomics of light-harvesting proteins in different plant species. Analysis and comparison by liquid chromatography-electrospray ionization mass spectrometry. Photosystem II. AB - An overview of the intact molecular masses and the hydrophobic properties of the photosystem II (PSII) light-harvesting proteins in 14 different plant species is presented. The protein separation and identification was achieved by means of reversed-phase high-performance liquid chromatography-electrospray ionization mass spectrometry. The good correspondence of the molecular masses measured by reversed-phase high-performance liquid chromatography-electrospray ionization mass spectrometry with those deduced from the DNA sequence (0.008%-0.016% relative deviation in Arabidopsis) enabled the identification of the different protein types. Utilizing this correlation, it was possible in several cases to spot a gene product for the previously cloned genes. In PSII, all antenna proteins show hydrophobic properties considerably different within the same as well as among various species, in contrast to observations made previously with PSI. These differences might reflect a tuning of protein-protein interactions that play a role in inducing different supramolecular organizations of PSII: within the same species as a consequence of short-term adaptations, and among species for seasonal species adaptation. The relative antenna stoichiometry was readily established on the basis of relative peak areas of the separated proteins in the ultraviolet chromatograms. The correspondence found between the high copy number of genes with the gene products reveals that the genes are not silent in their protein expression. Moreover, the high copy number of gene products as well as protein heterogeneity observed in PSII suggest a possible plant strategy to realize the high degree of organization and interconnection of the light harvesting systems under any environmental conditions. PMID- 12529529 TI - Alanine aminotransferase homologs catalyze the glutamate:glyoxylate aminotransferase reaction in peroxisomes of Arabidopsis. AB - Plant peroxisomal glyoxylate aminotransferases play central roles within the photorespiratory pathway. Genes encoding glyoxylate aminotransferases have been isolated from several animals and microbes, but only recently have plant homologs been identified. Three Arabidopsis homologs of alanine (Ala):glyoxylate aminotransferase 2 (AGT2) contain a putative type 1 peroxisomal targeting signal (PTS1), but the metabolic significance of these AGT2 homologs is unknown. GGT1 and GGT2 are Ala aminotransferase (AlaAT) homologs from Arabidopsis that represent another type of glyoxylate aminotransferase. These proteins are class I aminotransferases, each containing a putative PTS1. GGT1 and GGT2 are members of a small family of AlaATs in Arabidopsis. When expressed as recombinant proteins in Escherichia coli, GGT1 and GGT2 displayed biochemical characteristics very similar to one another, and to the Arabidopsis protein purified from leaves. Four aminotransferase activities were specifically associated with GGT1 and GGT2, using the substrate pairs glutamate (Glu):glyoxylate, Ala:glyoxylate, Glu:pyruvate, and Ala:2-oxoglutarate. GGT1 and GGT2 may have partially redundant functions; transcripts of both genes were detected in many of the same tissues. Although Glu:glyoxylate aminotransferase (GGT) activity has been observed in several locations in different plants and algae, including the cytoplasm and mitochondria, our subcellular fractionation data indicate that GGT activity was exclusively peroxisomal in Arabidopsis. Thus, glyoxylate aminotransferase reactions in plant peroxisomes appear to be catalyzed by at least two distinct types of aminotransferases: an AGT1 homolog with serine:glyoxylate aminotransferase activity (A.H. Liepman, L.J. Olsen [2001] Plant J 25: 487-498), and a pair of closely related, potentially redundant AlaAT homologs with GGT activity. PMID- 12529530 TI - Metabolism of sugars in the endosperm of developing seeds of oilseed rape. AB - The sugars in the endosperm of a developing seed have many potential roles, including the supply of carbon to the developing embryo and controlling gene expression in it. Our understanding of their metabolism is, however, fragmentary and is confined to a very few species (especially Vicia spp.). To develop a quantitative understanding of the regulation of sugars in seeds of oilseed rape (Brassica napus), we measured relevant enzyme activities, the sizes of the pools of sugars in the liquid endosperm, and the flux of sugars from the endosperm into the embryo. The concentrations of hexose sugars in the liquid endosperm decreased, and sucrose (Suc) increased through development. The overall osmotic potential also fell. The timing of the changes was not precise enough to determine whether they signaled the onset of rapid accumulation of storage products. Changes in endosperm invertase activity were complex and quantitatively do not explain the changes in sugars. The embryo can metabolize hexose sugars in addition to Suc, and possibly at higher rates. Therefore, in addition to invertase, the growing embryo itself has a potential to influence the balance of sugars in the endosperm. The activity of Suc synthase in the embryo was greater than that of invertase during development. This observation and a higher activity of fructokinase than glucokinase in the embryo are both consistent with the embryo using Suc as a carbon source. PMID- 12529531 TI - Metabolic origin of carbon isotope composition of leaf dark-respired CO2 in French bean. AB - The carbon isotope composition (delta(13)C) of CO(2) produced in darkness by intact French bean (Phaseolus vulgaris) leaves was investigated for different leaf temperatures and during dark periods of increasing length. The delta(13)C of CO(2) linearly decreased when temperature increased, from -19 per thousand at 10 degrees C to -24 per thousand at 35 degrees C. It also progressively decreased from -21 per thousand to -30 per thousand when leaves were maintained in continuous darkness for several days. Under normal conditions (temperature not exceeding 30 degrees C and normal dark period), the evolved CO(2) was enriched in (13)C compared with carbohydrates, the most (13)C-enriched metabolites. However, at the end of a long dark period (carbohydrate starvation), CO(2) was depleted in (13)C even when compared with the composition of total organic matter. In the two types of experiment, the variations of delta(13)C were linearly related to those of the respiratory quotient. This strongly suggests that the variation of delta(13)C is the direct consequence of a substrate switch that may occur to feed respiration; carbohydrate oxidation producing (13)C-enriched CO(2) and beta oxidation of fatty acids producing (13)C-depleted CO(2) when compared with total organic matter (-27.5 per thousand). These results are consistent with the assumption that the delta(13)C of dark respired CO(2) is determined by the relative contributions of the two major decarboxylation processes that occur in darkness: pyruvate dehydrogenase activity and the Krebs cycle. PMID- 12529532 TI - RNA target sequences promote spreading of RNA silencing. AB - It is generally recognized that a silencing-inducing locus can efficiently reduce the expression of genes that give rise to transcripts partially homologous to those produced by the silencing-inducing locus (primary targets). Interestingly, the expression of genes that produce transcripts without homology to the silencing-inducing locus (secondary targets) can also be decreased dramatically via transitive RNA silencing. This phenomenon requires primary target RNAs that contain sequences homologous to secondary target RNAs. Sequences upstream from the region homologous to the silencing inducer in the primary target transcripts give rise to approximately 22-nucleotide small RNAs, coinciding with the region homologous to the secondary target. The presence of these small RNAs corresponds with reduced expression of the secondary target whose transcripts are not homologous to the silencing inducer. The data suggest that in transgenic plants, targets of RNA silencing are involved in the expansion of the pool of functional small interfering RNAs. Furthermore, methylation of target genes in sequences without homology to the initial silencing inducer indicates not only that RNA silencing can expand across target RNAs but also that methylation can spread along target genes. PMID- 12529534 TI - Functional mitochondrial complex I is required by tobacco leaves for optimal photosynthetic performance in photorespiratory conditions and during transients. AB - The importance of the mitochondrial electron transport chain in photosynthesis was studied using the tobacco (Nicotiana sylvestris) mutant CMSII, which lacks functional complex I. Rubisco activities and oxygen evolution at saturating CO(2) showed that photosynthetic capacity in the mutant was at least as high as in wild type (WT) leaves. Despite this, steady-state photosynthesis in the mutant was reduced by 20% to 30% at atmospheric CO(2) levels. The inhibition of photosynthesis was alleviated by high CO(2) or low O(2). The mutant showed a prolonged induction of photosynthesis, which was exacerbated in conditions favoring photorespiration and which was accompanied by increased extractable NADP malate dehydrogenase activity. Feeding experiments with leaf discs demonstrated that CMSII had a lower capacity than the WT for glycine (Gly) oxidation in the dark. Analysis of the postillumination burst in CO(2) evolution showed that this was not because of insufficient Gly decarboxylase capacity. Despite the lower rate of Gly metabolism in CMSII leaves in the dark, the Gly to Ser ratio in the light displayed a similar dependence on photosynthesis to the WT. It is concluded that: (a) Mitochondrial complex I is required for optimal photosynthetic performance, despite the operation of alternative dehydrogenases in CMSII; and (b) complex I is necessary to avoid redox disruption of photosynthesis in conditions where leaf mitochondria must oxidize both respiratory and photorespiratory substrates simultaneously. PMID- 12529533 TI - Do phytotropins inhibit auxin efflux by impairing vesicle traffic? AB - Phytotropins such as 1-N-naphthylphthalamic acid (NPA) strongly inhibit auxin efflux, but the mechanism of this inhibition remains unknown. Auxin efflux is also strongly decreased by the vesicle trafficking inhibitor brefeldin A (BFA). Using suspension-cultured interphase cells of the BY-2 tobacco (Nicotiana tabacum L. cv Bright-Yellow 2) cell line, we compared the effects of NPA and BFA on auxin accumulation and on the arrangement of the cytoskeleton and endoplasmic reticulum (ER). The inhibition of auxin efflux (stimulation of net accumulation) by both NPA and BFA occurred rapidly with no measurable lag. NPA had no observable effect on the arrangement of microtubules, actin filaments, or ER. Thus, its inhibitory effect on auxin efflux was not mediated by perturbation of the cytoskeletal system and ER. BFA, however, caused substantial alterations to the arrangement of actin filaments and ER, including a characteristic accumulation of actin in the perinuclear cytoplasm. Even at saturating concentrations, NPA inhibited net auxin efflux far more effectively than did BFA. Therefore, a proportion of the NPA sensitive auxin efflux carriers may be protected from the action of BFA. Maximum inhibition of auxin efflux occurred at concentrations of NPA substantially below those previously reported to be necessary to perturb vesicle trafficking. We found no evidence to support recent suggestions that the action of auxin transport inhibitors is mediated by a general inhibition of vesicle-mediated protein traffic to the plasma membrane. PMID- 12529535 TI - Ethylene biosynthesis in detached young persimmon fruit is initiated in calyx and modulated by water loss from the fruit. AB - Persimmon (Diospyros kaki Thunb.) fruit are usually classified as climacteric fruit; however, unlike typical climacteric fruits, persimmon fruit exhibit a unique characteristic in that the younger the stage of fruit detached, the greater the level of ethylene produced. To investigate ethylene induction mechanisms in detached young persimmon fruit, we cloned three cDNAs encoding 1 aminocyclopropane-1-carboxylic acid (ACC) synthase (DK-ACS1, 2, and -3) and two encoding ACC oxidase (DK-ACO1 and -2) genes involved in ethylene biosynthesis, and we analyzed their expression in various fruit tissues. Ethylene production was induced within a few days of detachment in all fruit tissues tested, accompanied by temporally and spatially coordinated expression of all the DK-ACS and DK-ACO genes. In all tissues except the calyx, treatment with 1 methylcyclopropene, an inhibitor of ethylene action, suppressed ethylene production and ethylene biosynthesis-related gene expression. In the calyx, one ACC synthase gene (DK-ACS2) exhibited increased mRNA accumulation accompanied by a large quantity of ethylene production, and treatment of the fruit with 1 methylcyclopropene did not prevent either the accumulation of DK-ACS2 transcripts or ethylene induction. Furthermore, the alleviation of water loss from the fruit significantly delayed the onset of ethylene production and the expression of DK ACS2 in the calyx. These results indicate that ethylene biosynthesis in detached young persimmon fruit is initially induced in calyx and is modulated by water loss through transcriptional activation of DK-ACS2. The ethylene produced in the calyx subsequently diffuses to other fruit tissues and acts as a secondary signal that stimulates autocatalytic ethylene biosynthesis in these tissues, leading to a burst of ethylene production. PMID- 12529536 TI - Organ-specific expression of brassinosteroid-biosynthetic genes and distribution of endogenous brassinosteroids in Arabidopsis. AB - Brassinosteroids (BRs) are steroidal plant hormones that are essential for growth and development. There is only limited information on where BRs are synthesized and used. We studied the organ specificity of BR biosynthesis in Arabidopsis, using two different approaches: We analyzed the expression of BR-related genes using real-time quantitative reverse transcriptase-polymerase chain reaction, and analyzed endogenous BRs using gas chromatography-mass spectrometry. Before starting this study, we cloned the second BR-6-oxidase (BR6ox2) gene from Arabidopsis and found that the encoded enzyme has the same substrate specificity as the enzyme encoded by the previously isolated 6-oxidase gene (BR6ox1) of Arabidopsis. Endogenous BRs and the expression of BR-related genes were detected in all organs tested. The highest level of endogenous BRs and the highest expression of the BR6ox1, BR6ox2, and DWF4 genes were observed in apical shoots, which contain actively developing tissues. These genes are important in BR biosynthesis because they encode the rate-limiting or farthest downstream enzyme in the BR biosynthesis pathway. The second highest level of endogenous BRs and expression of BR6ox1 and DWF4 were observed in siliques, which contains actively developing embryos and seeds. These findings indicate that BRs are synthesized in all organs tested, but are most actively synthesized in young, actively developing organs. In contrast, synthesis was limited in mature organs. Our observations are consistent with the idea that BRs function as the growth promoting hormone in plants. PMID- 12529537 TI - Modulation of CYP79 genes and glucosinolate profiles in Arabidopsis by defense signaling pathways. AB - Glucosinolates are natural plant products that function in the defense toward herbivores and pathogens. Plant defense is regulated by multiple signal transduction pathways in which salicylic acid (SA), jasmonic acid, and ethylene function as signaling molecules. Glucosinolate content was analyzed in Arabidopsis wild-type plants in response to single or combinatorial treatments with methyljasmonate (MeJA), 2,6-dichloro-isonicotinic acid, ethylene, and 2,4 dichloro-phenoxyacetic acid, or by wounding. In addition, several signal transduction mutants and the SA-depleted transgenic NahG line were analyzed. In parallel, expression of glucosinolate biosynthetic genes of the CYP79 gene family and the UDPG:thiohydroximate glucosyltransferase was monitored. After MeJA treatment, the amount of indole glucosinolates increased 3- to 4-fold, and the corresponding Trp-metabolizing genes CYP79B2 and CYP79B3 were both highly induced. Specifically, the indole glucosinolate N-methoxy-indol-3 ylmethylglucosinolate accumulated 10-fold in response to MeJA treatment, whereas 4-methoxy-indol-3-ylmethylglucosinolate accumulated 1.5-fold in response to 2,6 dichloro-isonicotinic acid. In general, few changes were seen for the levels of aliphatic glucosinolates, although increases in the levels of 8-methylthiooctyl glucosinolate and 8-methylsulfinyloctyl glucosinolate were observed, particularly after MeJA treatments. The findings were supported by the composition of glucosinolates in the coronatine-insensitive mutant coi1, the ctr1 mutant displaying constitutive triple response, and the SA-overproducing mpk4 and cpr1 mutants. The present data indicate that different indole glucosinolate methoxylating enzymes are induced by the jasmonate and the SA signal transduction pathways, whereas the aliphatic glucosinolates appear to be primarily genetically and not environmentally controlled. Thus, different defense pathways activate subsets of biosynthetic enzymes, leading to the accumulation of specific glucosinolates. PMID- 12529538 TI - The binding of maize DHN1 to lipid vesicles. Gain of structure and lipid specificity. AB - Dehydrins (DHNs; late embryogenesis abundant D-11) are a family of plant proteins induced in response to abiotic stresses such as drought, low temperature, and salinity or during the late stages of embryogenesis. Spectral and thermal properties of these proteins in purified form suggest that they are "intrinsically unstructured." However, DHNs contain at least one copy of a consensus 15-amino acid sequence, the "K segment," which resembles a class A2 amphipathic alpha-helical, lipid-binding domain found in other proteins such as apolipoproteins and alpha-synuclein. The presence of the K segment raises the question of whether DHNs bind lipids, bilayers, or phospholipid vesicles. Here, we show that maize (Zea mays) DHN DHN1 can bind to lipid vesicles that contain acidic phospholipids. We also observe that DHN1 binds more favorably to vesicles of smaller diameter than to larger vesicles, and that the association of DHN1 with vesicles results in an apparent increase of alpha-helicity of the protein. Therefore, DHNs, and presumably somewhat similar plant stress proteins in the late embryogenesis abundant and cold-regulated classes may undergo function related conformational changes at the water/membrane interface, perhaps related to the stabilization of vesicles or other endomembrane structures under stress conditions. PMID- 12529539 TI - Divergent light-, ascorbate-, and oxidative stress-dependent regulation of expression of the peroxiredoxin gene family in Arabidopsis. AB - Peroxiredoxins (prxs) are peroxidases with broad substrate specificity. The seven prx genes expressed in Arabidopsis shoots were analyzed for their expressional response to changing photon fluence rates, oxidative stress, and ascorbate application. The results reveal a highly variable and gene-specific response to reducing and oxidizing conditions. The steady-state transcript amounts of the chloroplast-targeted prxs, namely the two-cysteine (2-Cys) prxs, prx Q and prx II E, decreased upon application of ascorbate. prx Q also responded to peroxides and diamide treatment. prx II B was induced by tertiary butylhydroperoxide, but rather unaffected by ascorbate. The strongest responses were observed for prx II C, which was induced with all treatments. The two Arabidopsis 2-Cys Prxs and four Prx II proteins were expressed heterologously in Escherichia coli. In an in vitro test system, they all showed peroxidase activity, but could be distinguished by their ability to accept dithiothreitol and thioredoxin as electron donor in the regeneration reaction. The midpoint redox potentials (E(m)') of Prx II B, Prx II C, and Prx II E were around -290 mV and, thus, less negative than E(m)' of Prx II F, 2-Cys Prx A, and 2-Cys Prx B (-307 to -322 mV). The data characterize expression and function of the mitochondrial Prx II F and the chloroplast Prx II E for the first time, to our knowledge. Antibodies directed against 2-Cys Prx and Prx II C showed a slight up-regulation of Prx II protein in strong light and of 2 Cys Prx upon transfer both to high and low light. The results are discussed in context with the subcellular localization of the Prx gene products. PMID- 12529540 TI - Sugar-regulated expression of a putative hexose transport gene in grape. AB - Different lengths of the promoter of grape (Vitis vinifera) VvHT1 (Hexose Transporter 1) gene, which encodes a putative hexose transporter expressed during the ripening of grape, have been transcriptionally fused to the beta glucuronidase reporter gene. In transgenic tobacco (Nicotiana tabacum) transformed with these constructs, VvHT1 promoters were clearly responsible for the sink organ preferential expression. The potential sugar effectors of VvHT1 promoter were studied in tobacco cv Bright-Yellow 2 cells transformed with chimeric constructs. Glucose (56 mM), sucrose (Suc; 58 mM), and the non transported Suc isomer palatinose doubled the beta-glucuronidase activity conferred by the VvHT1 promoter, whereas fructose did not affect it. These effects were the strongest with the 2.4-kb promoter, which contains all putative sugar-responsive elements (activating and repressing), but they were also significant with the 0.3-kb promoter, which contains only activating sugar boxes. The induction of VvHT1 expression by both Suc and palatinose was confirmed in the homologous grape berry cell culture. The data provide the first example of a putative sugar transporter, which is induced by both glucose and Suc in higher plants. Although induction of VvHT1 expression by Suc does not require transport, the presence of glucosyl moiety is necessary for Suc sensing. These results provide new insights into sugar sensing and signaling in plants. PMID- 12529541 TI - The pea gene NA encodes ent-kaurenoic acid oxidase. AB - The gibberellin (GA)-deficient dwarf na mutant in pea (Pisum sativum) has severely reduced internode elongation, reduced root growth, and decreased leaflet size. However, the seeds develop normally. Two genes, PsKAO1 and PsKAO2, encoding cytochrome P450 monooxygenases of the subfamily CYP88A were isolated. Both PsKAO1 and PsKAO2 had ent-kaurenoic acid oxidase (KAO) activity, catalyzing the three steps of the GA biosynthetic pathway from ent-kaurenoic acid to GA(12) when expressed in yeast (Saccharomyces cerevisiae). In addition to the intermediates ent-7alpha-hydroxykaurenoic acid and GA(12)-aldehyde, some additional products of the pea KAO activity were detected, including ent-6alpha,7alpha dihydroxykaurenoic acid and 7beta-hydroxykaurenolide. The NA gene encodes PsKAO1, because in two independent mutant alleles, na-1 and na-2, PsKAO1 had altered sequences and the five-base deletion in PsKAO1 associated with the na-1 allele cosegregated with the dwarf na phenotype. PsKAO1 was expressed in the stem, apical bud, leaf, pod, and root, organs in which GA levels have previously been shown to be reduced in na plants. PsKAO2 was expressed only in seeds and this may explain the normal seed development and normal GA biosynthesis in seeds of na plants. PMID- 12529542 TI - Quantitative trait loci analysis of nitrogen use efficiency in Arabidopsis. AB - Improving plant nitrogen (N) use efficiency or controlling soil N requires a better knowledge of the regulation of plant N metabolism. This could be achieved using Arabidopsis as a model genetic system, taking advantage of the natural variation available among ecotypes. Here, we describe an extensive study of N metabolism variation in the Bay-0 x Shahdara recombinant inbred line population, using quantitative trait locus (QTL) mapping. We mapped QTL for traits such as shoot growth, total N, nitrate, and free-amino acid contents, measured in two contrasting N environments (contrasting nitrate availability in the soil), in controlled conditions. Genetic variation and transgression were observed for all traits, and most of the genetic variation was identified through QTL and QTL x QTL epistatic interactions. The 48 significant QTL represent at least 18 loci that are polymorphic between parents; some may correspond to known genes from the N metabolic pathway, but others represent new genes controlling or interacting with N physiology. The correlations between traits are dissected through QTL colocalizations: The identification of the individual factors contributing to the regulation of different traits sheds new light on the relations among these characters. We also point out that the regulation of our traits is mostly specific to the N environment (N availability). Finally, we describe four interesting loci at which positional cloning is feasible. PMID- 12529543 TI - Polyamine metabolism is altered in unpollinated parthenocarpic pat-2 tomato ovaries. AB - Facultative parthenocarpy induced by the recessive mutation pat-2 in tomato (Lycopersicon esculentum Mill.) depends on gibberellins (GAs) and is associated with changes in GA content in unpollinated ovaries. Polyamines (PAs) have also been proposed to play a role in early tomato fruit development. We therefore investigated whether PAs are able to induce parthenocarpy and whether the pat-2 mutation alters the content and metabolism of PAs in unpollinated ovaries. Application of putrescine, spermidine, and spermine to wild-type unpollinated tomato ovaries (cv Madrigal [MA/wt]) induced partial parthenocarpy. Parthenocarpic growth of MA/pat-2 (a parthenocarpic near-isogenic line to MA/wt) ovaries was negated by paclobutrazol (GA biosynthesis inhibitor), and this inhibition was counteracted by spermidine. Application of alpha-difluoromethyl ornithine (-Orn) and/or alpha-difluoromethyl-arginine (-Arg), irreversible inhibitors of the putrescine biosynthesis enzymes Orn decarboxylase (ODC) and Arg decarboxylase, respectively, prevented growth of unpollinated MA/pat-2 ovaries. Alpha-difluoromethyl-Arg inhibition was counteracted by putrescine and GA(3), whereas that of alpha-difluoromethyl-Orn was counteracted by GA(3) but not by putrescine or spermidine. In unpollinated MA/pat-2 ovaries, the content of free spermine was significantly higher than in MA/wt ovaries. ODC activity was higher in pat-2 ovaries than in MA/wt. Transcript levels of genes encoding ODC and spermidine synthase were also higher in MA/pat-2. All together, these results strongly suggest that the parthenocarpic ability of pat-2 mutants depends on elevated PAs levels in unpollinated mutant ovaries, which correlate with an activation of the ODC pathway, probably as a consequence of elevated GA content in unpollinated pat-2 tomato ovaries. PMID- 12529544 TI - Exploring symbiotic nitrogen fixation and assimilation in pea root nodules by in vivo 15N nuclear magnetic resonance spectroscopy and liquid chromatography-mass spectrometry. AB - Nitrogen (N) fixation and assimilation in pea (Pisum sativum) root nodules were studied by in vivo (15)N nuclear magnetic resonance (NMR) by exposing detached nodules to (15)N(2) via a perfusion medium, while recording a time course of spectra. In vivo (31)P NMR spectroscopy was used to monitor the physiological state of the metabolically active nodules. The nodules were extracted after the NMR studies and analyzed for total soluble amino acid pools and (15)N labeling of individual amino acids by liquid chromatography-mass spectrometry. A substantial pool of free ammonium was observed by (15)N NMR to be present in metabolically active, intact nodules. The ammonium ions were located in an intracellular environment that caused a remarkable change in the in vivo (15)N chemical shift. Alkalinity of the ammonium-containing compartment may explain the unusual chemical shift; thus, the observations could indicate that ammonium is located in the bacteroids. The observed (15)N-labeled amino acids, glutamine/glutamate and asparagine (Asn), apparently reside in a different compartment, presumably the plant cytoplasm, because no changes in the expected in vivo (15)N chemical shifts were observed. Extensive (15)N labeling of Asn was observed by liquid chromatography-mass spectrometry, which is consistent with the generally accepted role of Asn as the end product of primary N assimilation in pea nodules. However, the Asn (15)N amino signal was absent in in vivo (15)N NMR spectra, which could be because of an unfavorable nuclear Overhauser effect. gamma-Aminobutyric acid accumulated in the nodules during incubation, but newly synthesized (15)N gamma aminobutyric acid seemed to be immobilized in metabolically active pea nodules, which made it NMR invisible. PMID- 12529545 TI - Stem cell molecular blueprint: "life, the universe, and everything". PMID- 12529546 TI - The unsolved enigmas of leukemia inhibitory factor. AB - Leukemia inhibitory factor (LIF) is a polyfunctional glycoprotein cytokine whose inducible production can occur in many, perhaps all, tissues. LIF acts on responding cells by binding to a heterodimeric membrane receptor composed of a low-affinity LIF-specific receptor and the gp130 receptor chain also used as the receptor for interleukin-6, oncostatin M, cardiotrophin-1, and ciliary neurotrophic factor. LIF is essential for blastocyst implantation and the normal development of hippocampal and olfactory receptor neurons. LIF is used extensively in experimental biology because of its key ability to induce embryonic stem cells to retain their totipotentiality. LIF has a wide array of actions, including acting as a stimulus for platelet formation, proliferation of some hematopoietic cells, bone formation, adipocyte lipid transport, adrenocorticotropic hormone production, neuronal survival and formation, muscle satellite cell proliferation, and acute phase production by hepatocytes. Unwanted actions of LIF can be minimized by circulating soluble LIF receptors and by intracellular suppression by suppressors of cytokine-signaling family members. However, the outstanding problems remain of how the induction of LIF is mediated in response to demands from such a heterogeneity of target tissues and why it makes design sense to use LIF in the regulation of such a diverse and unrelated series of biological processes. PMID- 12529547 TI - CD34- hematopoietic stem cells: current concepts and controversies. AB - Recent data have suggested that human CD34(-) hematopoietic stem cells (HSCs) exist, challenging the concept that HSCs necessarily and exclusively express the CD34 antigen. In mice, quiescent HSCs have been shown to be mostly CD34(-), but as a consequence of 5-fluorouracil treatment or cytokine stimulation, differentiate into CD34(+) cells. Of particular interest is a novel, specific marker to identify HSCs, namely the Hoechst dye efflux property, with which a distinct side population (SP) is identified. These SP cells are mostly CD34(-), highly enriched for long-term repopulating cells, and durably engraft in sublethally irradiated non-obese diabetic/severe combined immunodeficient mice. Using a semiquantitative reverse transcription-polymerase chain reaction, one of the ATP-binding cassette (ABC) transporters, the breast cancer resistance protein (Bcrp) or ABC transporter G2 (ABCG2), was found to be highly expressed in SP cells as well as other primitive HSCs and to sharply drop with hematopoietic differentiation. Enforced expression of the ABCG2 cDNA resulted in a robust SP phenotype and a reduction in hematopoietic maturation. These data suggest that the Bcrp/ABCG2 gene contributes importantly to the generation of the SP phenotype, which allows for the selection of immature, pluripotent HSCs. The isolation of Bcrp/ABCG2(+) cells appears to be an attractive tool to analyze and characterize HSCs, and may eventually allow for the purification of these cells for clinical purposes. In this review, current concepts on murine and human CD34( ) HSCs and their relationship with CD34(+) HSCs are discussed. PMID- 12529548 TI - Reprogramming immune responses: enabling cellular therapies and regenerative medicine. AB - Recent advances in cellular therapies have led to the emergence of a multidisciplinary scientific approach to developing therapeutics for a wide variety of diseases and genetic disorders. Although most cell-based therapies currently consist of heterogeneous cell populations, it is anticipated that the standard of care will eventually be well-characterized stem cell lines that can be modified to meet the individual needs of the patient. Many challenges have to be overcome, however, before such "designer cells" can become a clinical reality. One of the major hurdles will be to prevent immune rejection of the therapeutic cells. A patient's immune system may react to genetically modified or allogeneic cells as foreign, leading to their destruction. We propose that specific reprogramming of the immune system to accept cellular therapies can be accomplished by establishing hematopoietic chimerism. Successful engraftment of hematopoietic stem cells (HSCs), which have the same origin as those cells intended for therapeutic use, should lead to a re-education of the immune system so that the donor cells are recognized as self and will not be rejected. Developing safe, nontoxic protocols for reprogramming the immune system is critical to the success of this approach. Two major requirements exist for achieving stable HSC engraftment: (A) depletion or displacement of host stem cells, and (B) adequate immune suppression. Available data indicate that an agent such as busulfan is effective in depleting stem cells and that immune suppression can be accomplished with monoclonal antibodies that specifically target immune reactive cells in the periphery. PMID- 12529549 TI - The impact of progenitor enrichment, serum, and cytokines on the ex vivo expansion of mobilized peripheral blood stem cells: a controlled trial. AB - The aim of this study was to verify, and possibly improve, culture conditions to expand human mobilized peripheral blood stem cells (PBSCs). We investigated the role of three parameters: A) the culture medium (serum-free versus serum dependent); B) the initial cell population (Ficoll-separated mononucleated cells versus CD34(+)-selected cells), and C) the low concentration of recombinant cytokines, flt3 ligand, and thrombopoietin in association with a basic cocktail of stem cell factor, interleukin (IL)-6, IL-3, GM-CSF, and erythropoietin. Eighteen leukapheresis samples were monitored in static culture for 15 days. The expansion potential was assessed at day 10 and 15 by total nuclear cells, colony forming-units (CFUs) (burst-forming units-erythroid [BFU-E], colony-forming units granulocyte-macrophage [CFU-GM], and colony-forming units-granulocyte-erythroid macrophage-megakaryocyte [CFU-GEMM]), and flow cytometry immunophenotyping (CD34(+)/CD38(-), CD38(+), CD33(+), CD41(+), GlyA(+) progenitor cells). The results, evaluated by multivariate analysis of variance, emphasize that some variables affected the outcome of stem and progenitor cell expansion. CD34(+) enrichment increased expansion of total nuclear cells, number of CD38(+) and CD33(+) late precursors, and number of the CFU-GM compartment. Interestingly, however, quantitative expansion of GlyA(+) and the early progenitor cells (CD34(+)/CD38(-), CFU-GEMM, BFU-E) are favored by the use of unselected mononucleated cells. Regarding the role of serum, no significant difference was observed except for expansion of total nuclear cells, CFU-GM, and BFU-E. Cytokine combinations, in particular the use of flt3 ligand, stimulated expansion of almost all the cellular subsets, reaching a statistical significance for total nuclear cells and CFU-GM. Our study indicates that progenitor and late precursor multilineage cell compartments of mobilized PBSCs may be significantly expanded in short-term cultures by well-defined experimental conditions. Furthermore, these data might be useful when evaluating ex vivo expansion of hematopoietic cells for clinical purposes. PMID- 12529550 TI - A subset of ES-cell-derived neural cells marked by gene targeting. AB - Embryonic stem cells differentiate efficiently in culture into neural progenitors, neurons, oligodendrocytes, and astrocytes. An embryonic stem (ES) cell line with green fluorescent protein (GFP) inserted into the gene for Olig2, a lineage-specific transcription factor, permits visualization and physical separation of a subset of living ES-cell-derived neural cells. GFP-expressing cells have morphological and antigenic properties of the oligodendrocyte lineage. The differentiation of living GFP-expressing cells can be followed in cultures, and they can be separated by fluorescence-activated cell sorting and cultured as pure populations. This system will allow detailed biochemical and molecular analysis of a neural differentiation pathway at a level not previously feasible. The strategy may have general applicability, since other neural lineages can be marked in an analogous manner. PMID- 12529551 TI - Matrix cells from Wharton's jelly form neurons and glia. AB - We have identified an easily attainable source of primitive, potentially multipotent stem cells from Wharton's jelly, the matrix of umbilical cord. Wharton's jelly cells have been propagated in culture for more than 80 population doublings. Several markers for stem cells, including c-kit (CD117), and telomerase activity are expressed in these cells. Treatment with basic fibroblast growth factor overnight and low-serum media plus butylated hydroxyanisole and dimethylsulfoxide induced Wharton's jelly cells to express a neural phenotype. Within several hours of this treatment, Wharton's jelly cells developed rounded cell bodies with multiple neurite-like extensions, similar to the morphology of neural stem cells. Neuron-specific enolase (NSE), a neural stem cell marker, was expressed in these cells, as shown by immunocytochemistry. Immunoblot analysis showed similar levels of NSE expression in both untreated and induced Wharton's jelly cells. After 3 days, the induced Wharton's jelly cells resembled bipolar or multipolar neurons, with processes that formed networks reminiscent of primary cultures of neurons. The neuron-like cells in these cultures stained positively for several neuronal proteins, including neuron-specific class III beta-tubulin, neurofilament M, an axonal growth-cone-associated protein, and tyrosine hydroxylase. Immunoblot analysis showed increasing levels of protein markers for mature neurons over time post induction. Markers for oligodendrocytes and astrocytes were also detected in Wharton's jelly cells. These exciting findings show that cells from the matrix of umbilical cord have properties of stem cells and may, thus, be a rich source of primitive cells. This study shows their capacity to differentiate into a neural phenotype in vitro. PMID- 12529552 TI - Cytokine expansion culture of cord blood CD34+ cells induces marked and sustained changes in adhesion receptor and CXCR4 expressions. AB - Recent studies have demonstrated defective bone marrow homing of hematopoietic stem cells after cytokine expansion culture. Adhesion receptors (ARs) are essential to the homing process, and it is possible that cytokine culture modulates AR expression. We studied changes in expression of very late antigen-4 (VLA-4), VLA-5, L-selectin, leukocyte function-associated antigen-1 (LFA-1), CD44, and the stromal cell-derived factor-1 (SDF-1) receptor, CXCR4, during cytokine culture of cord blood (CB) CD34(+) cells. Expression of ARs was studied by flow cytometry on CB CD34(+) cells in whole blood, after purification and during culture for up to 10 days. Cells were cultured with stem cell factor (SCF), thrombopoietin (TPO), Flt3-ligand (Flt3), and G-CSF. Results showed that 80% or more of uncultured CD34(+) cells were positive for VLA-4, L-selectin, LFA 1, CD44, and CXCR4 while 50% were positive for VLA-5. Purification of CD34(+) cells did not affect AR expression, but cytokines increased expression three- to nine-fold throughout the 10-day culture period. In contrast, expression of CXCR4 decreased. Expression changes of ARs and CXCR4 on CD34(+)/CD38(-) cells mirrored those of the total CD34(+) population. The results indicate that cytokine culture significantly increases AR expression on CB CD34(+) cells, which may be related to the decrease in homing of cytokine-cultured hematopoietic stem cells. PMID- 12529554 TI - Hematopoietic recovery following autologous bone marrow transplantation in a nonhuman primate: effect of variation in treatment schedule with PEG-rHuMGDF. AB - Mathematical modeling of pegylated recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF) pharmacokinetics (PK) and pharmacodynamics (PD) suggest that variations in the PEG-rHuMGDF treatment schedule could reduce the severity and duration of thrombocytopenia following myeloablation and bone marrow transplant (BMT). We tested this hypothesis in a rhesus monkey model of autologous (Au) bone marrow-derived mononuclear cell (BM-MNC) transplantation following lethal myeloablation. On day 0, animals were myeloablated by total body exposure to 920 cGy, 250 kVp x-irradiation (TBI). Four cohorts of animals were infused with 1 x 10(8) AuBM-MNC/kg body weight within 2 hours of TBI. The AuBMT alone cohort received no cytokine, the daily dosage cohort received PEG-rHuMGDF (2.5 micro g/kg/day, s.c.) post TBI and AuBMT, and the pre/post-transplant cohort received PEG-rHuMGDF (2.5 micro g/kg/day, s.c.) pre (day -9 to day -5) and post TBI and AuBMT. The post-transplant PEG-rHuMGDF administration in the above cohorts was begun on day 1 post TBI and continued until platelet counts reached 200,000 micro l (range, 15-31 days). Another group received PEG-rHuMGDF (300 micro g/kg/day, s.c.) on days 1 and 3 only following TBI and AuBMT. The TBI controls received neither AuBMT nor cytokine therapy. In this model of AuBMT, with regard to the PEG-rHuMGDF administration schedule, the daily dosage of the post-transplant cohort did not significantly improve platelet recovery; the pre/post-transplant schedule and an abbreviated high-dosage, post-transplant schedule (days 1 and 3) significantly improved the duration and nadir of thrombocytopenia and platelet recovery. These data confirm predictions from PK/PD modeling of PEG-rHuMGDF that thrombocytopenia is preventable following AuBMT. PMID- 12529553 TI - Small-molecule-directed mpl signaling can complement growth factors to selectively expand genetically modified cord blood cells. AB - Efforts toward achieving gene therapy for blood disorders are plagued by low rates of gene transfer into hemopoietic stem cells. Recent studies suggest that this obstacle can be circumvented using selection. One way to achieve selection employs genes that encode receptor-bearing fusion proteins capable of inducing cell growth in response to drugs called chemical inducers of dimerization (CIDs). We have previously shown that genetically modified marrow cells from mice can proliferate for up to a year in culture in response to CID-initiated signals arising from the thrombopoietin receptor (mpl). The sustained growth observed in mouse hemopoietic cells results from an mpl-induced self-renewal of multipotential hemopoietic progenitor cells. In contrast, human hemopoietic cells proliferate only transiently in response to the mpl signal (from differentiation of transduced erythroid and megakaryocytic progenitors), while human myeloid progenitors fail to respond. Here, we show that myeloid progenitors from human cord blood can be induced to proliferate and/or differentiate in response to the mpl signal by providing additional signals via a combination of growth factors. These findings are relevant for the eventual clinical application of CID regulated cell therapy. PMID- 12529555 TI - Improved generation of germline-competent embryonic stem cell lines from inbred mouse strains. AB - Genetically altered mice may exhibit highly variable phenotypes due to the variation in genetic background, which can only be circumvented by generation of inbred, isogenic gene-targeted and control mice. Here we report that an embryonic stem (ES) cell culture medium conditioned by a rabbit fibroblast cell line transduced with genomic rabbit leukemia inhibitory factor allows efficient derivation and maintenance of ES cell lines from all of 10 inbred mouse strains tested, including some that were presumed to be nonpermissive for ES cell derivation (129/SvEv, 129/SvJ, C57BL/6N, C57BL/6JOla, CBA/CaOla, DBA/2N, DBA/1Ola, C3H/HeN, BALB/c, and FVB/N). Germline transmission was established by blastocyst injection of established ES cell lines after 10 or more passages from all of seven strains tested (129/SvJ, C57BL/6N, C57BL/6JOla, DBA/2N, DBA/1Ola, BALB/c, and FVB/N), by diploid aggregation of ES cell lines from all of four strains tested (129/SvEv, C57BL/6N, CBA/ CaOla, and FVB/N), or by tetraploid aggregation of ES cell lines from all of three strains tested (129/SvEv, C57BL/6N, and CBA/CaOla). Thus, these inbred ES cell lines may constitute useful tools to derive gene-targeted mice and isogenic controls in selected genetic backgrounds. PMID- 12529556 TI - Liver-specific gene expression in cultured human hematopoietic stem cells. AB - Hematopoietic and hepatic stem cells share characteristic markers such as CD34, c kit, and Thy1. Based on the recent observations that hepatocytes may originate from bone marrow, we investigated the potential of CD34(+) bone marrow cells to differentiate into hepatocytic cells in vitro. CD34(+) and CD34(-) human bone marrow cells were separated by magnetic cell sorting. Cells were cultured on a collagen matrix in a defined medium containing hepatocyte growth factor. Cell count and size were measured by flow cytometry, and reverse transcription polymerase chain reaction was carried out for the liver-specific markers CK-19 and albumin. During cell culture, CD34(+) cells showed an increasing cell number and proliferative activity as assessed by Ki-67 staining. Under the specified culture conditions, CD34(+) cells expressed albumin RNA and CK-19 RNA after 28 days, whereas CD34(-) cells did not show liver-specific gene expression. The results indicate that CD34(+) adult human bone marrow stem cells can differentiate into hepatocytic cells in vitro. PMID- 12529557 TI - Searching for alternative sources of postnatal human mesenchymal stem cells: candidate MSC-like cells from umbilical cord. AB - Mesenchymal stem cells (MSCs) have the capability for renewal and differentiation into various lineages of mesenchymal tissues. These features of MSCs attract a lot of attention from investigators in the context of cell-based therapies of several human diseases. Despite the fact that bone marrow represents the main available source of MSCs, the use of bone marrow-derived cells is not always acceptable due to the high degree of viral infection and the significant drop in cell number and proliferative/differentiation capacity with age. Thus, the search for possible alternative MSC sources remains to be validated. Umbilical cord blood is a rich source of hematopoietic stem/progenitor cells and does not contain mesenchymal progenitors. However, MSCs circulate in the blood of preterm fetuses and may be successfully isolated and expanded. Where these cells home at the end of gestation is not clear. In this investigation, we have made an attempt to isolate MSCs from the subendothelial layer of umbilical cord vein using two standard methodological approaches: the routine isolation of human umbilical vein endothelial cell protocol and culture of isolated cells under conditions appropriate for bone-marrow-derived MSCs. Our results suggest that cord vasculature contains a high number of MSC-like elements forming colonies of fibroblastoid cells that may be successfully expanded in culture. These MSC-like cells contain no endothelium- or leukocyte-specific antigens but express alpha smooth muscle actin and several mesenchymal cell markers. Therefore, umbilical cord/placenta stroma could be regarded as an alternative source of MSCs for experimental and clinical needs. PMID- 12529558 TI - High-level sustained transgene expression in human embryonic stem cells using lentiviral vectors. AB - Here we describe the sustained expression of transgenes introduced into human embryonic stem (ES) cells using self-inactivating lentiviral vectors. At low multiplicity of infection, vesicular stomatitis virus-pseudotyped vectors containing a green fluorescent protein (GFP) transgene under the control of a human elongation factor 1alpha promoter transduced human ES cells at high efficiency. The majority of the transduced ES cells, which harbored low numbers of integrated vectors, continued to express GFP after 60 days of culture. Incorporation of a scaffold attachment region (SAR) from the human interferon beta gene into the lentiviral vector backbone increased the average level of GFP expression, and inclusion of the SAR together with a chromatin insulator from the 5' end of the chicken beta-globin locus reduced the variability in GFP expression. When the transduced ES cells were induced to differentiate into CD34(+) hematopoietic precursors in vitro, GFP expression was maintained with minimal silencing. The ability to efficiently introduce active transgenes into human ES cells will facilitate gain-of-function studies of early developmental processes in the human system. These results also have important implications for the possible future use of gene-modified human ES cells in transplantation and tissue regeneration applications. PMID- 12529559 TI - The molecular perspective: VEGF and angiogenesis. PMID- 12529560 TI - Trends in mental health insurance benefits and out-of-pocket spending. AB - BACKGROUND: Insurance benefits can have a large effect on whether one is able to access health care services. Mental health and substance abuse (MHSA) insurance coverage has typically been less generous than that of general health services. AIMS OF THE STUDY: This paper examines trends in the generosity of private insurance benefits for mental health (MH) services in the United States from 1987 to 1996. The paper estimates the benefit-induced change in insurance payments for MH services that would have been made by typical health plans between 1987 and 1996 holding constant utilization of individuals at the 1987 level so that the changes in effective benefits could be isolated. METHODS: Trends in mental health benefits were measured using two nationally representative household surveys of the U.S. civilian non-institutionalized population, the 1987 National Medical Expenditure Survey (NMES) and the 1996 Medical Expenditure Panel Survey (MEPS). Data on utilization and expenditures from the NMES/MEPS were used to simulate what the average person would have paid out-of-pocket under typical insurance plans in 1987 and in 1996. RESULTS: The study finds that limits on MH coverage, such as limits on reimbursed days of care, became more prevalent from 1987 to 1996, but that consumer cost-sharing rates declined. The simulations indicate that private insurance would have paid for a lower proportion of total spending in 1996 (60.1 percent) as compared to 1987 (65.8 percent). DISCUSSION: Despite the fact that limits on mental health services became more prevalent over the time-period evaluated, out-of-pocket expenditures did not increase as significantly because there was a corresponding increase in coinsurance covered by health plans. IMPLICATIONS FOR HEALTH CARE PROVISION AND USE: Trends in plan design negatively affected those with high costs who are likely to surpass their limits and positively affected coverage for those with minimal use due to lower cost-sharing. These trends also indicate that persons in the most need, those with high utilization, particularly of inpatient care, experienced a decline in coverage while those with less intensive needs may have experienced a slight increase. IMPLICATIONS FOR HEALTH POLICIES: Out-of-pocket spending in both years of the study was substantial suggesting that improved health care coverage, such as that mandated in parity legislation, could improve access to care for persons needing mental health treatment. IMPLICATION FOR FURTHER RESEARCH: Additional research is needed to understand how trends in out-of-pocket spending and insurance benefits have influenced access to care. PMID- 12529562 TI - Quality standards and incentives in managed care organizations' specialty contracts for behavioral health. AB - BACKGROUND: In the US, most privately insured individuals are enrolled with managed care organizations (MCOs), and a majority of these organizations have subcontracted responsibility for behavioral health care to specialized vendors. Based on economic theory, we anticipate that MCOs should be more likely to require quality standards in contracts that transfer all financial risk to the vendor. AIMS OF THE STUDY: To test whether use of quality standards in behavioral health subcontracts differs between MCOs that transfer full financial risk and other MCOs. Similarly, to test for differences between for-profit and nonprofit MCOs. METHODS: Bivariate tests and logistic regression analysis of the use of five quality-related standards, and the use of any standard, in a nationally representative sample of commercial MCO products in 60 US market areas. Statistical controls include MCO size, chain affiliation, region and market size. RESULTS: All five standards we examined were widely used in behavioral health subcontracts (varying from 47% to 70% of products). However, contrary to our hypothesis, the standards are not more commonly used by MCO products with unlimited capitated contracts for behavioral health. In most cases the opposite is true. In addition, for-profit plans were more rather than less likely to use several of the standards. DISCUSSION: MCOs that transfer full risk may be using mechanisms other than quality standards (e.g. periodic rebidding) to prevent skimping; may be less concerned about quality anyway; or may be more skeptical about the value of existing standards. The fact that for-profit plans are equally or more likely to use these standards may reveal that their objectives are not different from those of nonprofits, or that competition is constraining them to adopt standards anyway. Limitations of this study include the lack of more detailed data on the nature of financial risk-sharing, and on the types of financial penalties associated with each standard. IMPLICATIONS FOR HEALTH POLICY: Pressure for accreditation appears to be an effective vehicle for encouraging the spread of standards. It would be useful to know how far use of these quality standards in contracts is linked to better quality of care. IMPLICATIONS FOR FUTURE RESEARCH: Further studies should examine the relationship between quality standards and quality of care PMID- 12529563 TI - Service organisation, service use and costs of community mental health care. AB - BACKGROUND: Different forms of service organisation may be expected to affect costs. I is important to understand the distribution of costs amongst providers of mental health care, so that the effect of service developments on budgets can be better anticipated and planned. AIMS: This study explored the association between different forms of mental health service organisation and costs. It tested the impact on costs of services with high degrees of integration between health and social care providers, and of services targeting at people with more severe mental health problems. METHODS: 260 service users in 4 districts in the north of England were interviewed and costs identified for each person. The districts comprised examples of four types of service configuration. Use and non use was compared by type of service, and after suitable transformations, costs were also compared between districts and between types of service. The sample was large enough to discern a difference of 50 ( 80) per week with 80% power and 5% significance. RESULTS: Costs were closely related to severity. People in targeted services had higher mean costs ( 136, SD 191 versus 92, SD 106; p=0.001). The costs of targeted services in this study were very similar to those of psychosis only services in a London study. Integrated services were predicted to have lower inpatient costs (p=0.003), lower PCG costs (p=0.003) and lower total health and social care costs (p=0.024). Differences between use and costs of specific services were largely attributable to imprecise definitions and supply-side factors. DISCUSSION: While the sample is not representative, the a priori distinctions made in selecting the districts were reflected in service use and costs. This study suggests strongly that targeting services at people with more severe mental health problems seems likely to increase mean health and social care costs per service user. It also suggests that integration maybe associated with lower costs. Since both targeting and integration are increasingly common mental health policies, it is important to recognise that they may have opposing effects on costs. PMID- 12529564 TI - Science discovery in clinician-economist collaboration: legacy and future challenges. AB - BACKGROUND: 2002 Carl Taube Lecture at the NIMH Mental Health Economics Meeting. AIMS OF THE STUDY: To analyze the contribution and process of clinician/economist collaboration. METHODS: Personal scientific autobiography, using relationships with three economists as case examples. RESULTS: In joint efforts by clinicians and economists, clinicians bring an interest in case examples and in responding to unmet need, while economists bring structured analysis methods and respect for a societal perspective. Through mutual respect and discovery, both clinicians and economists can define unmet need in clinical and economic terms and help develop models and programs to improve clinical care, while maintaining a societal evaluation perspective. Key to scientific discovery is the principle that the emotions generated by data, such as hope and despair, need to be acknowledged and utilized rather than avoided or buried, provided that such feelings are used in a balanced manner in research. According to the author, collaboration helps maintain such a balance. DISCUSSION: Collaboration requires and builds trust, and improves the depth of research by combining different personal and disciplinary perspectives and strengths. Young investigators should be encouraged to explore collaboration and to consider their feelings in response to health and economic data as an important scientific and creative resource. PMID- 12529566 TI - An analysis of the diffusion of new antidepressants: variety, quality, and marketing efforts. AB - BACKGROUND: We are not aware of any published research that quantifies and compares the importance of effectiveness and side effects for pharmaceutical sales, and that simultaneously incorporates the impacts of marketing efforts on the diffusion of new pharmaceutical agents in the U.S. The overall level and market share success of the various selective serotonin reuptake inhibitors ( SSRIs ) relative to a representative older generation tricyclic (such as amitriptyline) provides a useful focus for studying such issues. AIMS OF STUDY: To model jointly the marketing and sales relationships of the SSRIs in the U.S., to quantify the extent to which marketing efforts are responsive to the availability of new scientific information accompanying changes in quality and increases in product variety, and in turn to assess how the new FDA indication approvals and the enhanced marketing initiatives involving product quality and variety affect sales of the SSRI and other novel antidepressants. METHODS: Quarterly US sales, price, quantity and marketing data 1988Q1-1997Q4 are taken from IMS Health for the eight new antidepressants introduced into the US during this time period. Measures of physician-perceived quality attributes of the antidepressants are drawn from Market Measures, Inc., a medical survey research firm. These data are used to construct measures of product quality (effectiveness and side effect profile), and attribute variety across all antidepressants. Multivariate regression methods are used in estimating parameters of a marketing efforts model, a sales demand model encompassing the aggregate of the newer antidepressants, and a product share model. Simulation methods are employed to quantify elasticities. RESULTS: Since 1988, and relative to amitriptyline, there has been only a rather modest increase in the perceived average effectiveness of the SSRIs and related products, but the side effect profiles have improved substantially. Variety measures for effectiveness show greater increases over time than do those for side effects. Marketing efforts respond to science-based events, such as new FDA indication approvals, and to effectiveness and side effect quality improvements. Total antidepressant sales are positively and significantly related to price reductions, increased marketing efforts, and the level and variety of side effect profiles involving antidepressants. The level and variety of effectiveness does not significantly affect total antidepressant sales. Order of entry effects are important in affecting product market shares, while marketing efforts and relative quality attributes (particularly a more favorable side effect profile) have positive and significant impacts on relative market shares. IMPLICATIONS FOR HEALTH CARE PROVISION AND USE: Since patient response to SSRIs and related products is idiosyncratic, greater product variety facilitates better matching of antidepressant with patient. Much of the growth of the SSRIs and related antidepressants since 1988 can be attributed to increased product attribute variety, to improved changes in side effect quality relative to that of the tricyclics, and to the marketing of those improvements. IMPLICATIONS FOR HEALTH POLICIES: Marketing efforts play an important role in diffusing product information. Marketing efforts increase considerably following FDA approval for indications other than depression, and also increase with the average effectiveness and the average side effect rating of the products. IMPLICATIONS FOR FURTHER RESEARCH: Whether the relatively minor role that perceived effectiveness has in affecting sales relative to perceived side effect profile is unique to antidepressants, or generalizes to other therapeutic classes, merits further examination. PMID- 12529567 TI - A comparison of methods to handle skew distributed cost variables in the analysis of the resource consumption in schizophrenia treatment. AB - BACKGROUND: Transformation of the dependent cost variable is often used to solve the problems of heteroscedasticity and skewness in linear ordinary least square regression of health service cost data. However, transformation may cause difficulties in the interpretation of regression coefficients and the retransformation of predicted values. AIMS OF THE STUDY: The study compares the advantages and disadvantages of different methods to estimate regression based cost functions using data on the annual costs of schizophrenia treatment. METHODS: Annual costs of psychiatric service use and clinical and socio demographic characteristics of the patients were assessed for a sample of 254 patients with a diagnosis of schizophrenia (ICD-10 F 20.0) living in Leipzig. The clinical characteristics of the participants were assessed by means of the BPRS 4.0, the GAF, and the CAN for service needs. Quality of life was measured by WHOQOL-BREF. A linear OLS regression model with non-parametric standard errors, a log-transformed OLS model and a generalized linear model with a log-link and a gamma distribution were used to estimate service costs. For the estimation of robust non-parametric standard errors, the variance estimator by White and a bootstrap estimator based on 2000 replications were employed. Models were evaluated by the comparison of the R2 and the root mean squared error (RMSE). RMSE of the log-transformed OLS model was computed with three different methods of bias-correction. The 95% confidence intervals for the differences between the RMSE were computed by means of bootstrapping. A split-sample-cross-validation procedure was used to forecast the costs for the one half of the sample on the basis of a regression equation computed for the other half of the sample. RESULTS: All three methods showed significant positive influences of psychiatric symptoms and met psychiatric service needs on service costs. Only the log- transformed OLS model showed a significant negative impact of age, and only the GLM shows a significant negative influences of employment status and partnership on costs. All three models provided a R2 of about.31. The Residuals of the linear OLS model revealed significant deviances from normality and homoscedasticity. The residuals of the log-transformed model are normally distributed but still heteroscedastic. The linear OLS model provided the lowest prediction error and the best forecast of the dependent cost variable. The log-transformed model provided the lowest RMSE if the heteroscedastic bias correction was used. The RMSE of the GLM with a log link and a gamma distribution was higher than those of the linear OLS model and the log-transformed OLS model. The difference between the RMSE of the linear OLS model and that of the log-transformed OLS model without bias correction was significant at the 95% level. As result of the cross validation procedure, the linear OLS model provided the lowest RMSE followed by the log-transformed OLS model with a heteroscedastic bias correction. The GLM showed the weakest model fit again. None of the differences between the RMSE resulting form the cross- validation procedure were found to be significant. DISCUSSION: The comparison of the fit indices of the different regression models revealed that the linear OLS model provided a better fit than the log-transformed model and the GLM, but the differences between the models RMSE were not significant. Due to the small number of cases in the study the lack of significance does not sufficiently proof that the differences between the RSME for the different models are zero and the superiority of the linear OLS model can not be generalized. The lack of significant differences among the alternative estimators may reflect a lack of sample size adequate to detect important differences among the estimators employed. Further studies with larger case number are necessary to confirm the results. IMPLICATIONS: Specification of an adequate regression models requires a careful examination of the characteristics of the data. Estimation of standard errors and confidence intervals by nonparametric methods which are robust against deviations from the normal distribution and the homoscedasticity of residuals are suitable alternatives to the transformation of the skew distributed dependent variable. Further studies with more adequate case numbers are needed to confirm the results. PMID- 12529568 TI - Costs and effectiveness of substance abuse treatments for homeless persons. AB - BACKGROUND: Several reviews of the effectiveness of drug abuse treatment have concluded that treatment works. However, studies analyzing cost-effectiveness or cost-benefits of drug treatment have been limited. Consequently, policy decisions regarding substance abuse treatment have utilized educated guesses or consensus of experts in the absence of controlled and scientifically rigorous studies of the benefits and costs of treatment. AIMS OF THE STUDY: This study presents a cost analysis of two randomized controlled studies comparing four drug addiction interventions for homeless persons. The studies controlled for some limitations of previous research in this area including random assignment. Findings are based on treatment costs obtained from actual expenditures and treatment outcomes of drug abstinence from toxicology tests. Cost-effectiveness is considered from the viewpoint of the treatment program. Cost-effectiveness from a societal viewpoint is discussed, but not calculated. METHODS: This is a retrospective analysis of treatment and treatment outcome costs from two randomized controlled drug addiction treatment outcome studies: Homeless 1 and Homeless 2. Both studies were two-group-usual versus enhanced-care designs with similar treatment components, outcome variables and assessment points, but different research questions. Both studies investigated the efficacy of a contingency management intervention specifically designed for persons who are homeless. This costs analysis reports direct costs of treatment by service category and costs of abstinence at 2-, 6-, and 12-month points by study and study treatment group. Treatment costs and costs per week abstinent are reported for four substance abuse treatments across two studies for persons homeless and addicted primarily to crack cocaine. Treatment components for each program included counseling, housing, work, administrative, and other expenses. RESULTS: Drug abstinence rates by treatment program for each study revealed superior outcomes for the enhanced interventions with the greatest abstinence found at the earlier time points (up to 6 months) as established by previous research. Abstinence rates at 12 months failed to differentiate treatment groups. Average costs per abstinent week were generally greater for the enhanced programs compared to usual care, except early in treatment where these were similar. The incremental direct cost ratios (in year 2000 dollars) for these enhanced programs to increase abstinence by one average week were similar ($1,244 and $1,007) for the Homeless 1 and 2 projects at 12-months. These figures are compared to figures of other life saving events. DISCUSSION: When only the direct costs of programs and their abstinence rates are considered, treatments that involve abstinent contingent work and housing have incremental cost ratios that are within the range of many other common social and medical interventions. These enhanced programs are more cost effective earlier in treatment than at 12 month follow-up due to relapse common among existing drug treatment. A methodological limitation of this study is that direct program costs do not measure the societal value of reducing homelessness itself. IMPLICATIONS FOR HEALTH POLICIES: Usual and improved treatment methods offer a cost-effective approach to improving abstinence among addicted homeless persons. Policy makers might reasonably choose to implement enhanced treatment programs that also reduce homelessness because the incremental cost of these programs is within a reasonable range compared to other common societal interventions. IMPLICATIONS FOR FURTHER RESEARCH: Methods and data need to be developed to better measure the societal benefit to communities of reducing the numbers of homeless persons with addictive drug problems. PMID- 12529569 TI - Impact of primary care depression intervention on employment and workplace conflict outcomes: is value added? AB - BACKGROUND: Depression causes significant functional impairment in sufferers and often leads to adverse employment outcomes for working individuals. Recovery from depression has been associated with better employment outcomes at one year. AIMS OF THE STUDY: The study s goals were to assess a primary care depression intervention s impact on subsequent employment and workplace conflict outcomes in employed patients with depression. METHODS: In 1996-1997, the study enrolled 262 employed patients with depression from twelve primary care practices located across ten U.S. states; 219 (84%) of the patients were followed at one year. Intent-to-treat analyses assessing intervention effects on subsequent employment and workplace conflict were conducted using logistic regression models controlling for individual clinical and sociodemographic characteristics, job classification and local employment conditions. To meet criteria for subsequent employment, persons working full-time at baseline had to report they were working full-time at follow-up and persons working part-time at baseline had to report working part-/full-time at follow-up. Workplace conflict was measured by asking patients employed at follow-up whether, in the past year, they had arguments or other difficulties with people at work . RESULTS: Findings showed that 92.1% of intervention patients met criteria for subsequent employment at one year, versus 82.0% of usual care patients (c2=4.42, p=.04). Intervention patients were less likely than usual care patients to report workplace conflict in the year following baseline (8.1% vs. 18.9%, respectively; c2=4.11; p=.04). The intervention s effect on subsequent employment was not mediated by its effect on workplace conflict. DISCUSSION: The intervention significantly improved employment outcomes and reduced workplace conflict in depressed, employed persons at one year. Economic implications for employers related to reduced turnover costs, for workers related to retained earnings, and for governments related to reduced unemployment expenditures and increased tax receipts may be considerable. LIMITATIONS: Although similar primary care depression interventions have been shown to produce comparable effects on subsequent employment at one year, replications in larger samples of depressed, employed patients in different economic climates may be necessary to increase the generalizability and precision of estimates. IMPLICATIONS FOR HEALTH CARE PROVISION AND USE: Primary care interventions that enhance depression treatment and improve clinical outcomes can contribute meaningful added value to society by improving employment and workplace outcomes. IMPLICATIONS FOR HEALTH POLICIES: Federal/state governments may realize economic benefits from reduced unemployment expenditures and increased tax receipts should primary care depression interventions that improve employment outcomes be broadly disseminated. Policy initiatives to increase the dissemination of such interventions may be an innovative approach for improving labor force participation by depressed individuals. IMPLICATIONS FOR FURTHER RESEARCH: Formal cost-benefit analyses are needed to explore whether economic benefits to societal stakeholders from these and other labor outcomes equal or exceed the incremental costs of disseminating similar primary care interventions nationally. Researchers in other nations may wish to consider investigating the impact primary care depression interventions might have on employment and workplace outcomes in their countries. PMID- 12529570 TI - Auditory brain stem responses in the detection of brain death. AB - OBJECTIVES: We evaluated comatose patients by auditory brain stem responses (ABR) to determine the role of ABR in the diagnosis of impending brain death. PATIENTS AND METHODS: Sixty comatose patients in the intensive care unit were evaluated by brain stem evoked response audiometry. Correlations were sought between the absence or presence of ABRs and the presenting pathology, the Glasgow Coma Scale (GCS) scores, and ultimate diagnoses. RESULTS: The brain stem responses were totally absent in 41 patients. Presence of wave I could be obtained in only 10 patients. All the waveforms were found in nine patients; however, in eight patients the potentials disappeared as the GCS scores decreased to 3. Detection of wave I alone strongly suggested dysfunction of the brain stem. However, loss of wave I particularly in trauma patients aroused doubt as to whether the absence was associated with auditory end organ injury or brain stem dysfunction. CONCLUSION: The results suggest that evaluation of ABR may support brain death in a comatose patient (i) when wave I is present alone, (ii) the absence of wave I is accompanied by a documented auditory end organ injury, or (iii) when previously recorded potentials are no longer detectable. PMID- 12529571 TI - Carotid body tumors: challenging complexity of diagnosis and surgical treatment. AB - OBJECTIVES: To evaluate the patients treated for carotid body tumors and the problems encountered during diagnosis and treatment. PATIENTS AND METHODS: The study included eight patients (5 females, 3 males; mean age 50 years; range 36 to 68 years) who underwent surgery for carotid body tumors. Diagnosis was confirmed by power Doppler ultrasonography and computed tomography or magnetic resonance imaging in all patients. Angiography was performed in three patients to evaluate vascularity and to perform embolization before surgery. Two patients had previously undergone biopsy elsewhere. The mean follow-up was 15 months (range 2 to 36 months). RESULTS: The size of the tumors varied from 3 cm to 11 cm. In two patients the tumors were found to extend to the skull base. Ligation of the external carotid artery and the common carotid artery was performed in five patients and in one patient, respectively. Surgery-associated injuries given to the internal carotid artery in two patients were repaired by sutures. The 10th cranial nerve was dissected in two patients who had undergone a prior biopsy. They developed cord paralysis and hoarseness postoperatively. Resection of the 11th cranial nerve in one patient resulted in shoulder pain and drop shoulder. The 12th cranial nerve was repaired end-to-end by neurorrhaphy in two patients. They exhibited significant improvement in nerve functions a year after surgery. CONCLUSION: The larger the tumor is, the more difficult the resection is, and the more injuries are caused to the surrounding nerves and vessels. PMID- 12529572 TI - [Evaluation of the combined suprahyoid-transoral approach to the tongue base tumors]. AB - OBJECTIVES: To evaluate the efficacy and functional results of the combined suprahyoid-transoral approach in early-stage tongue base cancers. PATIENTS AND METHODS: Five patients (4 females, 1 male; mean age 47 years; range 12 to 65 years) with tongue base tumors underwent surgery via the combined suprahyoid transoral approach. Diagnoses were adenoid cystic carcinoma in three patients and squamous cell carcinoma in two patients. The mean follow-up period was 32.4 months (range 10 to 54 months). RESULTS: One patient with adenoid cystic carcinoma and two patients with squamous cell carcinoma required neck dissection. Postoperative radiotherapy was administered to two patients with squamous cell carcinoma. Tracheotomy was performed in all cases during surgery. All patients were safely decannulated within a mean of 11.4 days (range 9 to 14 days) and converted to oral feeding within a mean of 8.6 days (range 7 to 11 days) with no aspiration problem. All patients had negative surgical margins. All patients had satisfactory phonation. No local or regional recurrences developed during the follow-up period. CONCLUSION: Functional results obtained through the combined suprahyoid-transoral approach justify its use in the treatment of T1 or T2 tongue base carcinomas. PMID- 12529573 TI - [Histologic effects of hyperbaric oxygen therapy administered immediately after or two hours after ischemia-reperfusion injury: a rat abdominal skin flap model]. AB - OBJECTIVES: In a rat abdominal skin flap model, we investigated the histologic effects of hyperbaric oxygen (HBO) therapy on the skin flaps and the lung tissue immediately after and two hours after ischemia-reperfusion injury (IRI). STUDY DESIGN: Thirty Sprague-Dawley rats were assigned to three groups equal in number. The first group was left as controls. The left inferior abdominal skin flap based on the superficial inferior epigastric vessels was elevated. Following 10 hours of global ischemia, HBO therapy was started immediately after and two hours after reperfusion in groups 2 and 3, respectively. Our protocol included a course of 16 HBO treatments in six days at 2.5 ATA for 90 minutes, with four sessions on the first day, three sessions on the second and third days, and two sessions on the fourth, fifth, and sixth days. On the sixth day, survival rates for all flaps were calculated. Histologic examination was made on specimens taken from rat flaps and lungs. RESULTS: The mean flap survival was 25.87% in the control group. It significantly increased to 90.93% and 84.34% in HBO-treated rats in groups 2 and 3, respectively (p<0.001). There was no significant difference between HBO treated groups (p>0.05). Histologically, neither minor nor major changes were observed in HBO-treated rat lung tissues. Similarly, compared to the controls, thrombosis in flap microvasculature, intimal damage, edema, neutrophil infiltration, and necrosis were considerably less and neovascularization was significantly higher in HBO-treated flap tissues. CONCLUSION: Intense HBO treatment protocol used in this study significantly increases flap survival in rat axial skin flap model and reduces deleterious effects of IRI. PMID- 12529574 TI - [Landau-Kleffner syndrome (acquired epileptic aphasia)]. AB - Landau-Kleffner syndrome is characterized by a complex group of symptoms including deterioration in language skills, seizures, and abnormal electroencephalography findings. A six-year-old male patient had aphasia for three years and generalized tonic-clonic epileptic seizures for two years. Pure tone audiometry and auditory brain-stem response audiometry findings were normal. He had verbal auditory agnosia rather than true aphasia. Cranial computed tomography and magnetic resonance imaging studies did not show any abnormal findings. Single-photon emission computed tomography showed hypoperfusion in the right hemispheric temporal lobe. Despite treatment with corticosteroids and intravenous immunoglobulins, multiple exacerbations were interspersed in a four year follow-up period. PMID- 12529575 TI - Ectopic thyroid and Hashimoto's thyroiditis arising from a thyroglossal duct cyst: a case report. AB - A seventy-five-year-old male patient presented with a palpable smooth mass that moved upwards on swallowing, extending from the hyoid bone to the cricoid cartilage. Ultrasonography, scintigraphy, and thyroid hormone measurements showed the mass free from the thyroid gland. Following a diagnosis of infected thyroglossal duct cyst by fine needle aspiration biopsy, the mass was removed by surgery. Histopathologic diagnosis was Hashimoto's thyroiditis that developed from the ectopic thyroid tissue on the wall of thyroglossal duct cyst. PMID- 12529576 TI - [Reconstruction of the anomalous palatoglossal arch causing dysphagia: a case report]. AB - Physical examination of a 20-year-old male patient suffering from dysphagia revealed an abnormally short and medially displaced palatoglossal arch. The mucosa was released by an inverse V incision, and the palatoglossal muscle was found to be located more laterally and cephalad than its usual insertion, attached to the posterior edge of the hard palate. The muscle was transposed medially and caudally to its normal localization on the soft palate, and the palatoglossal arch was reconstructed by closure of the mucosal incision in an inverse Y manner. The patient's complaint disappeared and no complications were observed in the postoperative period. PMID- 12529577 TI - Pulmonary thromboembolism following total laryngectomy and neck dissection: a case report. AB - A fifty-five-year-old male patient underwent total laryngectomy, bilateral modified radical neck dissection, and primary voice restoration for squamous cell carcinoma of the supraglottic larynx. During surgery the left internal jugular vein was found to be thrombosed and, therefore, ligated and resected. In the early postoperative period, pulmonary thromboembolism was suspected and confirmed by lung perfusion scintigraphy which showed bilateral segmental and subsegmental perfusion defects. The patient was successfully treated by anticoagulant therapy. It may be advisable to consider thromboprophylaxis in head and neck surgery in patients with clinically suspected pulmonary thromboembolism, if no contraindication exists. PMID- 12529578 TI - [Nitric oxide and the upper airways: recent discoveries]. AB - Nitric oxide (NO) is produced from L-arginine in mammalian tissues. Nitric oxide synthase (NOS) catalyzes this reaction in human tissues. It has been shown that NO serves as an important signaling molecule in the cardiovascular system and is responsible for vasoregulation. In 1991, NO was discovered in exhaled air. More recently, it has been shown that the main production site of exhaled NO is the nose and sinuses. In the upper airways NO upregulates ciliary motility and provides a first-line defense mechanism against microorganisms by antiviral and antimicrobial activity. In the lungs it is involved in ventilation/perfusion matching. Nitric oxide is also a marker for inflammation, with increased nasal output in allergic rhinitis, and a decreased output in sinusitis, nasal polyps and Kartagener's syndrome. This report reviews some aspects of the origin, metabolism, and functions of NO in the upper airways, together with the techniques for, and implications of, nasal NO measurement. PMID- 12529579 TI - A measurement system for rhinoplasty patients. PMID- 12529580 TI - [Mild cognitive decline in the elderly: nosology and clinical status]. AB - Cognitive impairment without dementia is common in elderly persons and causes significant impairment in capacity to perform everyday activities. A number of nosological entities have been proposed for the classification of sub-clinical cognitive dysfunction. The more recent concepts assume an underlying organic condition, in particular the presence of early stage dementia. While prospective studies suggest that persons with mild cognitive disorder have a high risk of developing dementia, long-term follow-up of subjects with mild cognitive impairment suggests that dementia alone does not explain all cases. Cognitive disorder in the elderly must be construed as a common outcome for a number of interacting pathologies whose expression is also mediated by genetic, environmental and social factors. PMID- 12529581 TI - [Limits and uncertainties about the concept of MCI: contribution of neuropsychological approach]. AB - The diagnosis of Alzheimer disease is earlier with the memory clinics at a stage preceding the DSMIV criterions of dementia when the complaints are only in the field of memory without daylife impairment (stage CDR 0,5 at the Morris clinical dementia rating). This "predemential" stage is obviously more uncertain in terms of diagnosis and prognosis than the later one but some groups declare the same reliability with combining neuropsychology, biological markers in CSF and imagery. The MCI as a pure syndromic definition but the prospective studies have demonstrated that only 20 to 30p. cent of MCI will remain steady after 5 years of following, and several autopsies of MCI have showed that the neuropathological frontier of Alzheimer disease were crossed with neurofibrillary tangles and senile plaques in the hippocampus. Many trials have been conducted in the different fields of neuropsychology, biological markers and imagery to distinguish the MCI "decliners" from the MCI not related to Alzheimer. In that paper we will analyze all the clinical and neuropsychological perspectives to make that distinction with the maximum of reliability. The question of the usefulness of the MCI concept remains open, taking into account the progress in the very early stage diagnosis of Alzheimer's disease. PMID- 12529582 TI - [Mild cognitive impairment: imaging data]. AB - The diagnosis of Mild Cognitive Impairment (MCI) as defined by Petersen et al. (1997), is based uniquely on clinical observations. Cerebral imaging, both morphological and functional, may in fact facilitate diagnosis, particularly with regard to the differentiation of sub-types of MCI and the identification of prodromal AD (MCI-AD). Volumetric Magnetic Resonance Imagery (MRI) examination of structures affected early in AD such as entorhinal cortex, the temporal lobe and, above all, the hippocampus, are especially useful. Hypoactivity within these regions, especially of the temporal lobe and posterior cingulate gyrus by Positon Emission Tomography, and more recently monophotonic emission tomoscintigraphy, also appears to have diagnostic utility. Studies of cholinergic system activity by functional imaging may also be of future value in MCI-AD diagnosis. MRI used in conjunction with other techniques may be of significant value, in particular the use of spectro-MRI and functional MRI. This latter technique, leading to the development of cognitive activation paradigms, is particularly promising. PMID- 12529583 TI - [Mild Cognitive Impairment or pre-demential Alzheimer's disease?]. AB - The concept of Mild Cognitive Impairment (MCI) is proposed to define a state of cognition where the deficiency is greater than expected for a subject's age and socio-cultural background, but not sufficiently severe to satisfy the criteria of nosographic classifications of dementia. The concept of MCI cannot provide an etiological diagnosis but can be useful for recognizing and following patients with mild impairment. It raises the question of early diagnosis of Alzheimer's disease. The risk of progression to dementia is greater in patients with MCI, particularly those whose memory is the only domain involved - "amnesic MCI". Patients with amnesic MCI can be distinguished from those with MCI with multiple domains slightly impaired or non-amnesic MCI where only one domain (language, visual-spatial.) other than memory is involved. Depending on the type of deficiency, amnesic MCI may progress to different types of disease states. At the early stage of pure memory impairment, a distinction should be made between Alzheimer-related memory disorders and those related to another neurological conditions or the brain aging process. The question is whether the diagnosis of Alzheimer's disease can be made at this stage. Biological markers and brain imaging provide useful information, but these diagnostic tools remain imprecise and have not been validated for routine use. The process of diagnosis must therefore focus on an analysis of the cognitive impairment. The predementia phase of Alzheimer's disease is characterized by a progressive "hippocampal" decline in memory, sometimes associated with impaired execution. A careful analysis of the cognitive impairment helps identify "hippocampal amnesia syndrome" suggestive of the diagnosis of Alzheimer's disease. PMID- 12529584 TI - [Mild Cognitive Impairment: potential therapeutics]. AB - Mild Cognitive Impairment (MCI) is an emerging concept used to describe memory decline and probably attention disturbances in otherwise intellectually intact individuals. MCI may be considered in 12 to 15 p. 100 of the cases as announcing an Alzheimer's Disease (AD). Although still speculative, the debate concerning the drugs susceptible to normalize symptoms of MCI or to stop conversion to AD must be raised. For that purpose, several long term clinical trials are running (antioxidants, nootropics, anticholinesterasics.) and new molecules in the pipe line should be assessed in patients with the diagnosis of MCI. PMID- 12529586 TI - The crisis in nursing. PMID- 12529587 TI - From uninformed patient to CWOCN: My life with ulcerative colitis and the ileoanal reservoir. PMID- 12529588 TI - Tools of outcome measurement in WOCN practice. PMID- 12529589 TI - What interventions are effective for the prevention and treatment of cutaneous candidiasis? PMID- 12529590 TI - Skin temperature and chronic venous insufficiency. AB - OBJECTIVE: This study compared lower leg skin temperatures in adults age 50 years and older with and without chronic venous insufficiency (CVI). Norms and ranges of skin temperature were also established. DESIGN: A descriptive comparative design was used. SETTING AND SUBJECTS: Sixty-six adults recruited from hospital clinics and senior community dwellings participated in the study, which was conducted in an examination room of an outpatient clinic. To be included in the study, those with CVI showed stage 4 and 5 clinical signs. Those without CVI had no clinical signs. INSTRUMENTS: Skin temperature was measured with an infrared dermal thermometer. METHODS: Subjects were seated with legs extended, free of clothing and socks, and loosely covered with a draped sheet for 10 minutes to acclimatize to room temperature. The skin temperature of 4 sites on the lower legs was measured. Analysis of variance and independent t tests were used to compare groups. RESULTS: There was a significant increase in skin temperature at all 4 sites in the ankle area in the group with CVI compared with the comparison group (89.9 degrees F +/- 2.7 degrees F versus 88.1 degrees F +/- 2.1 degrees F, P =.004). CONCLUSIONS: Elevated skin temperature may be an important nursing assessment parameter to evaluate worsening or impending CVI complications such as venous ulcers. PMID- 12529591 TI - The views of district nurses on their level of knowledge about the treatment of leg and foot ulcers. AB - OBJECTIVE: The aim was to survey the views of a sample of district nurses on their own level of knowledge about the treatment of leg and foot ulcers and what they considered to be the most important issues in wound treatment. DESIGN: A descriptive design was used. SETTING AND SUBJECTS: The subjects were all the nurses who treated leg and foot ulcers in the community health care service in Oslo, Norway, during autumn 1999. The subjects were selected after consultation with senior nurses. INSTRUMENTS: A structured questionnaire with blank spaces for in-depth comments was used. METHODS: The head of the nursing service in all 25 urban districts received a letter about the study with a questionnaire that they passed on to the nurses treating ulcers. Out of 173 eligible nurses, 102 (59%) filled in the questionnaires. Descriptive and interpretative analyses were then made. RESULTS: Most of the participating nurses (60%) believed that their knowledge of wound treatment was insufficient. A high degree of uncertainty related to wound treatment situations was also found. The uncertainty included wound treatment, wound assessment, wound healing products, and wound causes. The main sources of the nurses' knowledge were their own nursing practice and colleagues. Twenty different factors of importance in wound treatment were reported. The most frequently mentioned factors were: wound healing, continuity in the treatment process, alleviation of pain, hygiene, compression therapy, and diagnosis. The lowest scores were awarded to documentation and economic factors. CONCLUSIONS: The nurses perceived their knowledge of leg and foot ulcer treatment to be insufficient. Their own nursing practice and colleagues were their main sources of knowledge. PMID- 12529592 TI - Meanings and practical knowledge of people with long-term urinary catheters. AB - The lived experience of people with long-term indwelling urethral catheters is described and interpreted. This hermeneutic (interpretive) phenomenologic study used Merleau-Ponty's philosophy and van Manen's methodology. Fourteen adults aged 35 to 95 years who had worn a catheter more than 4 months (range = 6 months to 18 years) comprised the purposive sample. Data were obtained through audiotaped and transcribed interviews conducted in the homes of participants. The study provided both meanings and practical knowledge related to the experience of catheter use. Despite its drawbacks, participants recognized the device's value in their daily lives and weighed its positive features against concerns associated with long term use. Individual meanings included making aesthetic harmony with a new catheter, the convenience of the catheter compared with the inconvenience of incontinence, and the catheter as a symbol and reminder of one's mortality. Practical knowledge related to participants' awareness of their catheter needs and practices, such as features of supplies, intervals for changes, catheter insertions, emptying the urine bag, and changes associated with sex. Knowledge of the lived experience with a urinary catheter can assist nurses in sensitive decision making about care and help them become better patient advocates. PMID- 12529593 TI - Pelvic floor muscle exercises and manometric biofeedback for erectile dysfunction and postmicturition dribble: three case studies. AB - Three successful case studies of men receiving treatment for erectile dysfunction and postmicturition dribble are presented to alert nurses to the possible benefits of pelvic floor muscle exercises. PMID- 12529594 TI - Patient with chronic venous stasis ulcer. PMID- 12529596 TI - More heat, less light on Lomborg. PMID- 12529595 TI - Climate come-uppance delayed. PMID- 12529597 TI - US officials urge biologists to vet publications for bioterror risk. PMID- 12529598 TI - Gravity experiment sparks spat between physicists. PMID- 12529599 TI - Safety doubts force rethink of embattled comet mission. PMID- 12529600 TI - Report backs Smithsonian research. PMID- 12529601 TI - German researchers set to receive Israeli stem-cell shipment. PMID- 12529602 TI - Prospect of human cloning poses dilemma for journals. PMID- 12529603 TI - Paris university blasted over Israel motion. PMID- 12529605 TI - Ethics panel attacks environment book. PMID- 12529604 TI - Pesticide firms ask to use human data to assess safety. PMID- 12529607 TI - Burning issues. PMID- 12529608 TI - Optical atomic clocks: The times, they are a-changin'. PMID- 12529609 TI - Journals: how to decide what's worth publishing. PMID- 12529610 TI - Journals: redundant publications are bad news. PMID- 12529611 TI - Journals: impact factors are too highly valued. PMID- 12529612 TI - Bright students enjoy correcting the textbooks. PMID- 12529613 TI - Animal research needs organized defence. PMID- 12529614 TI - DNA discrepancy. PMID- 12529619 TI - Science in culture: A mechanical mind. PMID- 12529621 TI - Microbial phylogenomics: Branching out. PMID- 12529622 TI - Sugars tied to the spot. PMID- 12529623 TI - Functional genomics: RNA sets the standard. PMID- 12529624 TI - Palaeoclimatology: Cooling a continent. PMID- 12529626 TI - Quantum cryptography: Code-breakers confounded. PMID- 12529625 TI - Immunology: Mobilizing the army. PMID- 12529628 TI - Evolutionary biology: Splitting in space. PMID- 12529629 TI - Developmental biology: Germ-cell attraction. PMID- 12529631 TI - Vulcanology: Interaction between Kilauea and Mauna Loa. PMID- 12529632 TI - Cell biology: Tubulin acetylation and cell motility. PMID- 12529633 TI - Evolution of the Archaean crust by delamination and shallow subduction. AB - The Archaean oceanic crust was probably thicker than present-day oceanic crust owing to higher heat flow and thus higher degrees of melting at mid-ocean ridges. These conditions would also have led to a different bulk composition of oceanic crust in the early Archaean, that would probably have consisted of magnesium-rich picrite (with variably differentiated portions made up of basalt, gabbro, ultramafic cumulates and picrite). It is unclear whether these differences would have influenced crustal subduction and recycling processes, as experiments that have investigated the metamorphic reactions that take place during subduction have to date considered only modern mid-ocean-ridge basalts. Here we present data from high-pressure experiments that show that metamorphism of ultramafic cumulates and picrites produces pyroxenites, which we infer would have delaminated and melted to produce basaltic rocks, rather than continental crust as has previously been thought. Instead, the formation of continental crust requires subduction and melting of garnet-amphibolite--formed only in the upper regions of oceanic crust--which is thought to have first occurred on a large scale during subduction in the late Archaean. We deduce from this that shallow subduction and recycling of oceanic crust took place in the early Archaean, and that this would have resulted in strong depletion of only a thin layer of the uppermost mantle. The misfit between geochemical depletion models and geophysical models for mantle convection (which include deep subduction) might therefore be explained by continuous deepening of this depleted layer through geological time. PMID- 12529636 TI - Quantum key distribution using gaussian-modulated coherent states. AB - Quantum continuous variables are being explored as an alternative means to implement quantum key distribution, which is usually based on single photon counting. The former approach is potentially advantageous because it should enable higher key distribution rates. Here we propose and experimentally demonstrate a quantum key distribution protocol based on the transmission of gaussian-modulated coherent states (consisting of laser pulses containing a few hundred photons) and shot-noise-limited homodyne detection; squeezed or entangled beams are not required. Complete secret key extraction is achieved using a reverse reconciliation technique followed by privacy amplification. The reverse reconciliation technique is in principle secure for any value of the line transmission, against gaussian individual attacks based on entanglement and quantum memories. Our table-top experiment yields a net key transmission rate of about 1.7 megabits per second for a loss-free line, and 75 kilobits per second for a line with losses of 3.1 dB. We anticipate that the scheme should remain effective for lines with higher losses, particularly because the present limitations are essentially technical, so that significant margin for improvement is available on both the hardware and software. PMID- 12529635 TI - Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. AB - A principal challenge currently facing biologists is how to connect the complete DNA sequence of an organism to its development and behaviour. Large-scale targeted-deletions have been successful in defining gene functions in the single celled yeast Saccharomyces cerevisiae, but comparable analyses have yet to be performed in an animal. Here we describe the use of RNA interference to inhibit the function of approximately 86% of the 19,427 predicted genes of C. elegans. We identified mutant phenotypes for 1,722 genes, about two-thirds of which were not previously associated with a phenotype. We find that genes of similar functions are clustered in distinct, multi-megabase regions of individual chromosomes; genes in these regions tend to share transcriptional profiles. Our resulting data set and reusable RNAi library of 16,757 bacterial clones will facilitate systematic analyses of the connections among gene sequence, chromosomal location and gene function in C. elegans. PMID- 12529637 TI - Single-nanowire electrically driven lasers. AB - Electrically driven semiconductor lasers are used in technologies ranging from telecommunications and information storage to medical diagnostics and therapeutics. The success of this class of lasers is due in part to well developed planar semiconductor growth and processing, which enables reproducible fabrication of integrated, electrically driven devices. Yet this approach to device fabrication is also costly and difficult to integrate directly with other technologies such as silicon microelectronics. To overcome these issues for future applications, there has been considerable interest in using organic molecules, polymers, and inorganic nanostructures for lasers, because these materials can be fashioned into devices by chemical processing. Indeed, amplified stimulated emission and lasing have been reported for optically pumped organic systems and, more recently, inorganic nanocrystals and nanowires. However, electrically driven lasing, which is required in most applications, has met with several difficulties in organic systems, and has not been addressed for assembled nanocrystals or nanowires. Here we investigate the feasibility of achieving electrically driven lasing from individual nanowires. Optical and electrical measurements made on single-crystal cadmium sulphide nanowires show that these structures can function as Fabry-Perot optical cavities with mode spacing inversely related to the nanowire length. Investigations of optical and electrical pumping further indicate a threshold for lasing as characterized by optical modes with instrument-limited linewidths. Electrically driven nanowire lasers, which might be assembled in arrays capable of emitting a wide range of colours, could improve existing applications and suggest new opportunities. PMID- 12529638 TI - Rapid Cenozoic glaciation of Antarctica induced by declining atmospheric CO2. AB - The sudden, widespread glaciation of Antarctica and the associated shift towards colder temperatures at the Eocene/Oligocene boundary (approximately 34 million years ago) (refs 1-4) is one of the most fundamental reorganizations of global climate known in the geologic record. The glaciation of Antarctica has hitherto been thought to result from the tectonic opening of Southern Ocean gateways, which enabled the formation of the Antarctic Circumpolar Current and the subsequent thermal isolation of the Antarctic continent. Here we simulate the glacial inception and early growth of the East Antarctic Ice Sheet using a general circulation model with coupled components for atmosphere, ocean, ice sheet and sediment, and which incorporates palaeogeography, greenhouse gas, changing orbital parameters, and varying ocean heat transport. In our model, declining Cenozoic CO2 first leads to the formation of small, highly dynamic ice caps on high Antarctic plateaux. At a later time, a CO2 threshold is crossed, initiating ice-sheet height/mass-balance feedbacks that cause the ice caps to expand rapidly with large orbital variations, eventually coalescing into a continental-scale East Antarctic Ice Sheet. According to our simulation the opening of Southern Ocean gateways plays a secondary role in this transition, relative to CO2 concentration. PMID- 12529639 TI - Discovery of abundant hydrothermal venting on the ultraslow-spreading Gakkel ridge in the Arctic Ocean. AB - Submarine hydrothermal venting along mid-ocean ridges is an important contributor to ridge thermal structure, and the global distribution of such vents has implications for heat and mass fluxes from the Earth's crust and mantle and for the biogeography of vent-endemic organisms. Previous studies have predicted that the incidence of hydrothermal venting would be extremely low on ultraslow spreading ridges (ridges with full spreading rates <2 cm x yr(-1)-which make up 25 per cent of the global ridge length), and that such vent systems would be hosted in ultramafic in addition to volcanic rocks. Here we present evidence for active hydrothermal venting on the Gakkel ridge, which is the slowest spreading (0.6-1.3 cm x yr(-1)) and least explored mid-ocean ridge. On the basis of water column profiles of light scattering, temperature and manganese concentration along 1,100 km of the rift valley, we identify hydrothermal plumes dispersing from at least nine to twelve discrete vent sites. Our discovery of such abundant venting, and its apparent localization near volcanic centres, requires a reassessment of the geologic conditions that control hydrothermal circulation on ultraslow-spreading ridges. PMID- 12529640 TI - Increased CO2 uncouples growth from isoprene emission in an agriforest ecosystem. AB - The emission of isoprene from the leaves of forest trees is a fundamental component of biosphere-atmosphere interactions, controlling many aspects of photochemistry in the lower atmosphere. As almost all commercial agriforest species emit high levels of isoprene, proliferation of agriforest plantations has significant potential to increase regional ozone pollution and enhance the lifetime of methane, an important determinant of global climate. Here we show that growth of an intact Populus deltoides plantation under increased CO2 (800 micromol x mol(-1) and 1,200 micromol x mol(-1)) reduced ecosystem isoprene production by 21% and 41%, while above-ground biomass accumulation was enhanced by 60% and 82%, respectively. Exposure to increased CO2 significantly reduced the cellular content of dimethylallyl diphosphate, the substrate for isoprene synthesis, in both leaves and leaf protoplasts. We identify intracellular metabolic competition for phosphoenolpyruvate as a possible control point in explaining the suppression of isoprene emission under increased CO2. Our results highlight the potential for uncoupling isoprene emission from biomass accumulation in an agriforest species, and show that negative air-quality effects of proliferating agriforests may be offset by increases in CO2. PMID- 12529641 TI - Speciation along environmental gradients. AB - Traditional discussions of speciation are based on geographical patterns of species ranges. In allopatric speciation, long-term geographical isolation generates reproductively isolated and spatially segregated descendant species. In the absence of geographical barriers, diversification is hindered by gene flow. Yet a growing body of phylogenetic and experimental data suggests that closely related species often occur in sympatry or have adjacent ranges in regions over which environmental changes are gradual and do not prevent gene flow. Theory has identified a variety of evolutionary processes that can result in speciation under sympatric conditions, with some recent advances concentrating on the phenomenon of evolutionary branching. Here we establish a link between geographical patterns and ecological processes of speciation by studying evolutionary branching in spatially structured populations. We show that along an environmental gradient, evolutionary branching can occur much more easily than in non-spatial models. This facilitation is most pronounced for gradients of intermediate slope. Moreover, spatial evolutionary branching readily generates patterns of spatial segregation and abutment between the emerging species. Our results highlight the importance of local processes of adaptive divergence for geographical patterns of speciation, and caution against pitfalls of inferring past speciation processes from present biogeographical patterns. PMID- 12529642 TI - Loss and recovery of wings in stick insects. AB - The evolution of wings was the central adaptation allowing insects to escape predators, exploit scattered resources, and disperse into new niches, resulting in radiations into vast numbers of species. Despite the presumed evolutionary advantages associated with full-sized wings (macroptery), nearly all pterygote (winged) orders have many partially winged (brachypterous) or wingless (apterous) lineages, and some entire orders are secondarily wingless (for example, fleas, lice, grylloblattids and mantophasmatids), with about 5% of extant pterygote species being flightless. Thousands of independent transitions from a winged form to winglessness have occurred during the course of insect evolution; however, an evolutionary reversal from a flightless to a volant form has never been demonstrated clearly for any pterygote lineage. Such a reversal is considered highly unlikely because complex interactions between nerves, muscles, sclerites and wing foils are required to accommodate flight. Here we show that stick insects (order Phasmatodea) diversified as wingless insects and that wings were derived secondarily, perhaps on many occasions. These results suggest that wing developmental pathways are conserved in wingless phasmids, and that 're evolution' of wings has had an unrecognized role in insect diversification. PMID- 12529643 TI - Genome-wide RNAi analysis of Caenorhabditis elegans fat regulatory genes. AB - Regulation of body fat storage involves signalling between centres that regulate feeding in the brain and sites of fat storage and use in the body. Here we describe an assay for analysing fat storage and mobilization in living Caenorhabditis elegans. By using RNA-mediated interference (RNAi) to disrupt the expression of each of the 16,757 worm genes, we have systematically screened the C. elegans genome for genes necessary for normal fat storage. We identify 305 gene inactivations that cause reduced body fat and 112 gene inactivations that cause increased fat storage. Analysis of the fat-reducing gene inactivations in insulin, serotonin and tubby signalling mutants of C. elegans, which have increased body fat, identifies a core set of fat regulatory genes as well as pathway-specific fat regulators. Many of the newly identified worm fat regulatory genes have mammalian homologues, some of which are known to function in fat regulation. Other C. elegans fat regulatory genes that are conserved across animal phylogeny, but have not previously been implicated in fat storage, may point to ancient and universal features of fat storage regulation, and identify targets for treating obesity and its associated diseases. PMID- 12529644 TI - Coupling of agonist binding to channel gating in the GABA(A) receptor. AB - Neurotransmitters such as acetylcholine and GABA (gamma-aminobutyric acid) mediate rapid synaptic transmission by activating receptors belonging to the gene superfamily of ligand-gated ion channels (LGICs). These channels are pentameric proteins that function as signal transducers, converting chemical messages into electrical signals. Neurotransmitters activate LGICs by interacting with a ligand binding site, triggering a conformational change in the protein that results in the opening of an ion channel. This process, which is known as 'gating', occurs rapidly and reversibly, but the molecular rearrangements involved are not well understood. Here we show that optimal gating in the GABA(A) receptor, a member of the LGIC superfamily, is dependent on electrostatic interactions between the negatively charged Asp 57 and Asp 149 residues in extracellular loops 2 and 7, and the positively charged Lys 279 residue in the transmembrane 2-3 linker region of the alpha1-subunit. During gating, Asp 149 and Lys 279 seem to move closer to one another, providing a potential mechanism for the coupling of ligand binding to opening of the ion channel. PMID- 12529645 TI - Periodic notch inhibition by lunatic fringe underlies the chick segmentation clock. AB - The segmented aspect of the vertebrate body plan first arises through the sequential formation of somites. The periodicity of somitogenesis is thought to be regulated by a molecular oscillator, the segmentation clock, which functions in presomitic mesoderm cells. This oscillator controls the periodic expression of 'cyclic genes', which are all related to the Notch pathway. The mechanism underlying this oscillator is not understood. Here we show that the protein product of the cyclic gene lunatic fringe (Lfng), which encodes a glycosyltransferase that can modify Notch activity, oscillates in the chick presomitic mesoderm. Overexpressing Lfng in the paraxial mesoderm abolishes the expression of cyclic genes including endogenous Lfng and leads to defects in segmentation. This effect on cyclic genes phenocopies inhibition of Notch signalling in the presomitic mesoderm. We therefore propose that Lfng establishes a negative feedback loop that implements periodic inhibition of Notch, which in turn controls the rhythmic expression of cyclic genes in the chick presomitic mesoderm. This feedback loop provides a molecular basis for the oscillator underlying the avian segmentation clock. PMID- 12529646 TI - SAP is required for generating long-term humoral immunity. AB - Long-lived plasma cells and memory B cells are the primary cellular components of long-term humoral immunity and as such are vitally important for the protection afforded by most vaccines. The SAP gene has been identified as the genetic locus responsible for X-linked lymphoproliferative disease, a fatal immunodeficiency. Mutations in SAP have also been identified in some cases of severe common variable immunodeficiency disease. The underlying cellular basis of this genetic disorder remains unclear. We have used a SAP knockout mouse model system to explore the role of SAP in immune responses. Here we report that mice lacking expression of SAP generate strong acute IgG antibody responses after viral infection, but show a near complete absence of virus-specific long-lived plasma cells and memory B cells, despite the presence of virus-specific memory CD4+ T cells. Adoptive transfer experiments show that SAP-deficient B cells are normal and the defect is in CD4+ T cells. Thus, SAP has a crucial role in CD4+ T-cell function: it is essential for late B-cell help and the development of long-term humoral immunity but is not required for early B-cell help and class switching. PMID- 12529647 TI - Responses of ferns to red light are mediated by an unconventional photoreceptor. AB - Efficient photosynthesis is essential for plant survival. To optimize photosynthesis, plants have developed several photoresponses. Stems bend towards a light source (phototropism), chloroplasts move to a place of appropriate light intensity (chloroplast photorelocation) and stomata open to absorb carbon dioxide. These responses are mediated by the blue-light receptors phototropin 1 (phot1) and phototropin 2 (phot2) in Arabidopsis (refs 1-5). In some ferns, phototropism and chloroplast photorelocation are controlled by red light as well as blue light. However, until now, the photoreceptor mediating these red-light responses has not been identified. The fern Adiantum capillus-veneris has an unconventional photoreceptor, phytochrome 3 (phy3), which is a chimaera of the red/far-red light receptor phytochrome and phototropin. We identify here a function of phy3 for red-light-induced phototropism and for red-light-induced chloroplast photorelocation, by using mutational analysis and complementation. Because phy3 greatly enhances the sensitivity to white light in orienting leaves and chloroplasts, and PHY3 homologues exist among various fern species, this chimaeric photoreceptor may have had a central role in the divergence and proliferation of fern species under low-light canopy conditions. PMID- 12529648 TI - Direct activation of RNA polymerase III transcription by c-Myc. AB - The proto-oncogene product c-Myc has a direct role in both metazoan cell growth and division. RNA polymerase III (pol III) is involved in the generation of transfer RNA and 5S ribosomal RNA, and these molecules must be produced in bulk to meet the need for protein synthesis in growing cells. We demonstrate here that c-Myc binds to TFIIIB, a pol III-specific general transcription factor, and directly activates pol III transcription. Chromatin immunoprecipitation reveals that endogenous c-Myc is present at tRNA and 5S rRNA genes in cultured mammalian cells. These results suggest that activation of pol III may have a role in the ability of c-Myc to stimulate cell growth. PMID- 12529651 TI - Chemistry's clouded view. PMID- 12529652 TI - Will industry play a bigger role in Europe's aims to promote mobility among postdocs? PMID- 12529653 TI - Possible dominant-negative mutation of the SHIP gene in acute myeloid leukemia. AB - The SH2 domain-containing inositol 5'-phosphatase (SHIP) is crucial in hematopoietic development. To evaluate the possible tumor suppressor role of the SHIP gene in myeloid leukemogenesis, we examined primary leukemia cells from 30 acute myeloid leukemia (AML) patients, together with eight myeloid leukemia cell lines. A somatic mutation at codon 684, replacing Val with Glu, was detected in one patient, lying within the signature motif 2, which is the phosphatase active site. The results of an in vitro inositol 5'-phosphatase assay revealed that the mutation reduced catalytic activity of SHIP. Leukemia cells with the mutation showed enhanced Akt phosphorylation following IL-3 stimulation. K562 cells transfected with the mutated SHIP-V684E cDNA showed a growth advantage even at lower serum concentrations and resistance to apoptosis induced by serum deprivation and exposure to etoposide. These results suggest a possible role of the mutated SHIP gene in the development of acute leukemia and chemotherapy resistance through the deregulation of the phosphatidylinositol-3,4,5 triphosphate (PI(3,4,5)P3)/Akt signaling pathway. This is the first report of a mutation in the SHIP gene in any given human cancer, and indicates the need for more attention to be paid to this gene with respect to cancer pathogenesis. PMID- 12529654 TI - New mechanisms of AML1 gene alteration in hematological malignancies. AB - The human AML1 gene (also named CBFA2 or RUNX1), located in the 21q22 chromosomal band, encodes for one of the two subunits forming a heterodimeric transcription factor, the human core binding factor (CBF). AML1 protein contains a highly evolutionary conserved domain of 128 amino acids called runt domain, responsible for both heterodimerization with the beta subunit of CBF and for DNA binding. AML1 is normally expressed in all hematopoietic lineages and acts to regulate the expression of various genes specific to hematopoiesis playing a pivotal role in myeloid differentiation. AML1 is one of the genes most frequently deregulated in leukemia through different mechanisms including translocation, mutation and amplification. Translocations lead to the formation of fusion genes encoding for chimerical proteins such as AML1-ETO which induces leukemogenesis. Recently, new mechanisms of AML1 deregulation by point mutations or amplification have been reported. To our knowledge, 51 patients (among 805 studied) with AML1 point mutations have been described. Forty of them have acute myeloid leukemia (AML) most often M0 AML. In this subtype of AML, the frequency of AML1 mutation is significantly higher; 21.5% of patients mutated (34/158). Mutations have also been found with lower frequency in other FAB subtype AML (6 cases), in myeloproliferative disorders (6 cases), in myelodysplastic syndrome (3 cases) and rarely in acute lymphoblastic leukemia (1 case). AML1 gene amplification has been found essentially in childhood ALL (12 cases) and more rarely in myeloid malignancies (4 cases). Here, we reviewed all these cases of AML1 point mutations and amplification and focused on the mechanisms of AML1 deregulation induced by these alterations. PMID- 12529655 TI - Molecular determinants of glucocorticoid sensitivity and resistance in acute lymphoblastic leukemia. AB - Glucocorticoids (GC) are probably the most important drugs in the treatment of ALL. Despite the extensive use of GC for many years, little is known about the molecular mechanisms of sensitivity and resistance. This review summarizes the knowledge on GC cytotoxicity in leukemia. The relevance of polymorphisms, splice variants and the number and regulation of the GC receptor are discussed. The role of multidrug resistance proteins, glutathione and glutathione S-transferase is evaluated, as well as the influence of the different heat-shock chaperone (hsp 90 and 70) and co-chaperone proteins (BAG-1 and others) which form a complex together with the GC receptor. Finally, the transactivation and transrepression (via NF-kappa B and AP-1 binding) of a wide range of genes (like c-myc) which initiates the final apoptosis pathway are discussed and suggestions for future directions of research in ALL patients are given. PMID- 12529656 TI - Molecular and cellular aspects of HTLV-1 associated leukemogenesis in vivo. AB - Most cancers and leukemias are preceded by a prolonged period of clinical latency during which cellular, chromosomal and molecular aberrations help move normal cell towards the malignant phenotype. The problem is that premalignant cells are usually indistinguishable from their normal counterparts, thereby ruling out the possibility to investigate the events that govern early leukemogenesis in vivo. Adult T cell leukemia/lymphoma (ATLL) is a T cell malignancy that occurs after a 40-60-year period of clinical latency in about 3-5% of HTLV-1-infected individuals. ATLL cells are monoclonally expanded and harbor an integrated provirus. A persistent oligo/polyclonal expansion of HTLV-1-bearing cells has been shown to precede ATLL, supporting the fact that in ATLL tumor cells arise from a clonally expanding non-malignant cell. It is possible to isolate infected, ie preleukemic, cells during the premalignant asymptomatic phase of the infection, thus providing an exceptional system to study the mechanisms underlying human cancers. Here we review some of the consequences of HTLV-1 on its host cell in vivo, at different stages of infection. PMID- 12529657 TI - High numbers of mobilized stem cells in patients with acute myeloid leukemia in first complete remission: good or bad? PMID- 12529658 TI - Immunomodulatory analogs of thalidomide inhibit growth of Hs Sultan cells and angiogenesis in vivo. AB - We have previously shown that thalidomide and its potent immunomodulatory derivatives (IMiDs) inhibit the in vitro growth of multiple myeloma (MM) cell lines and patient MM cells that are resistant to conventional therapy. In this study, we further characterize the effect of these drugs on growth of B cell malignancies and angiogenesis. We established a beige-nude-xid (BNX) mouse model to allow for simultaneous in vivo measurement of both anti-tumor and anti angiogenic effects of thalidomide and its analogs. Daily treatment (50 mg/kg/d) with thalidomide or IMiDs was nontoxic. The IMiDs were significantly more potent than thalidomide in vivo in suppressing tumor growth, evidenced by decreased tumor volume and prolonged survival, as well as mediating anti-angiogenic effects, as determined by decreased microvessel density. Our results therefore show that the IMiDs have more potent direct anti-tumor and anti-angiogenic effects than thalidomide in vivo, providing the framework for clinical protocols evaluating these agents in MM and other B cell neoplasms. PMID- 12529659 TI - Long-term outcome with pentostatin treatment in hairy cell leukemia patients. A French retrospective study of 238 patients. AB - With the introduction of new drugs such as alpha-interferon (IFN) and purine analogs, the management of hairy cell leukemia (HCL) patients has changed. However, deoxycoformycin (DCF) produced higher complete remission rates than IFN. The current study was undertaken to provide long-term data on duration of overall survival (OS) and disease-free survival (DFS) and incidence of subsequent malignancies. We retrospectively analyzed the data of patients treated with DCF (4 mg/m2/day, every 2 weeks) from 39 French centers. In 84 of 238 included patients, DCF was the first-line therapy. Pretreatment variables influencing the achievement of complete remission, DFS, and OS were identified by multivariate analysis. Two hundred and thirty-eight patients received a median of nine cycles (range, 1-19 cycles). A complete remission was obtained in 182 of 230 evaluable patients (79%) and a partial response was obtained in 38 patients, for an overall response rate of 95.6%. In the multivariate analysis hemoglobin level less than 100 g/l and leukocytes less than 2 x 10(9)/l were parameters adversely influencing complete remission achievement. With a median follow-up of 63.5 months (range, 0.39-138.4 months), disease recurrence was observed in 34 of 220 responding patients (15%). The estimated 5-years and 10-years DFS was 88.1% and 68.8%, respectively. Hemoglobin level less than 100 g/l and leukocytes less than 2 x 10(9)/l were the pre-treatment variables associated with a shorter DFS. The estimated 5-year and 10-year OS were 89.4% and 88.7%, respectively. Hemoglobin level less than 100 g/l, leukocytes less than 2 x 10(9)/l, and adenopathy were significant factors of reduced survival. Hematologic toxicity was the main side effect, followed by infection and emesis. During the period of follow-up, 18 patients developed second cancer, but the standardized incidence ratio was 0.95. Pentostatin is a highly effective regimen for hairy cell leukemia that produces durable complete responses. Toxicity of DCF is acceptable. Subsequent malignancies do not appear to be increased with pentostatin treatment. PMID- 12529660 TI - Aplidine, a new anticancer agent of marine origin, inhibits vascular endothelial growth factor (VEGF) secretion and blocks VEGF-VEGFR-1 (flt-1) autocrine loop in human leukemia cells MOLT-4. AB - The mechanism by which aplidine, a marine natural product in early clinical development as an anticancer agent, induces cell growth inhibition and apoptosis has been investigated in the human leukemia cell line MOLT-4. This cell line is characterized not only by the ability to secrete VEGF, but also for the presence on its surface of the VEGF receptor-1 (VEGFR-1). Previous studies from our laboratory concerned with evaluating early changes in gene expression induced by aplidine in MOLT-4 cells have shown that the drug decreases the expression of VEGFR-1 (Marchini et al. Proc Am Assoc Cancer Res 2000; 41: 833). Here, we report the ability of aplidine to block the VEGF/VEGFR-1 loop. We found that aplidine blocked VEGF secretion that was temporally followed by a decrease in both VEGF and VEGFR-1 production. Aplidine did not directly affect either VEGF transcription or stabilization of its mRNA. Transfection of MOLT-4 cells with an antisense VEGF cDNA construct, resulted in inhibition of colony formations. One clone, transfected with sense VEGF cDNA, secreting 8-10 times more VEGF than parental cells, was less sensitive to aplidine-induced cytotoxicity and apoptosis than control cells. Moreover, addition of VEGF in the medium decreased the activity of aplidine in MOLT-4 cells. These data demonstrate that aplidine inhibits the growth and induces apoptosis in MOLT-4 cells through the inhibition of VEGF secretion which blocks the VEGF/VEGFR-1 autocrine loop necessary for the growth of these cells. PMID- 12529661 TI - The stem cell mobilizing capacity of patients with acute myeloid leukemia in complete remission correlates with relapse risk: results of the EORTC-GIMEMA AML 10 trial. AB - Variable numbers of CD34+ cells can be harvested from the blood of AML patients in CR after G-CSF supported mobilization following consolidation chemotherapy. We hypothesized that a decreased ability to mobilize stem cells reflects a chemotherapy-induced reduction in the number of normal and leukemic stem cells. We therefore analyzed whether the mobilizing capacity of these patients was of prognostic significance. 342 AML-patients in first CR received daily G-CSF from day 20 of the consolidation course and underwent 1-6 aphereses to obtain a minimum dose of 2 x 10(6) CD34+ cells/kg. Afterwards they were randomized for autologous bone marrow (BM) or blood SCT. As a surrogate marker for the mobilizing capacity, the highest yield of CD34+ cells of a single apheresis was adopted. Patients could be categorized into four groups: no harvest (n = 76), low yield (<1 x 10(6) CD34+/kg; n = 50), intermediate yield (1-6.9 x 10(6) CD34+ cells/kg; n = 128) and high yield (> or = 7 x 10(6) CD34+ cells/kg; n = 88). The median follow-up was 3.4 years; 163 relapses and 16 deaths in CR were reported. Autologous blood or BM SCT was performed in 36%, 64%, 81% and 88%, respectively, of the patients assigned to the no harvest, low, intermediate and high CD34+ yield group. The 3-year disease-free survival rate was 46.7%, 65.0%, 50.4% and 26.9% (P = 0.0002) and the relapse incidence was 47.5%, 30.1%, 43.1% and 71.9% (P < 0.0001). Multivariate Cox's proportional hazards model showed that the CD34+ yield was the most important independent prognostic variable (P = 0.005) after cytogenetics. Patients with the highest mobilizing capacity have a poor prognosis due to an increased relapse incidence. PMID- 12529662 TI - High percentage of CD34-positive cells in autologous AML peripheral blood stem cell products reflects inadequate in vivo purging and low chemotherapeutic toxicity in a subgroup of patients with poor clinical outcome. AB - In this study, a high CD34% in autologous peripheral blood stem cell (PBSC) products from 71 AML patients was associated directly with a high relapse rate (P = 0.006) and inversely with disease-free survival (P = 0.003), irrespective whether patients were transplanted or not. The relapse rate at 12 months was 67% in a group with >0.8% CD34+ cells and 34% in a group with < or = 0.8% CD34+ cells. Although the percentage of malignant CD34+ cells in the CD34+ compartment in the relapses of the first group was not high (median 8%), the total number of malignant cells as a percentage of WBC was about 13 times higher than for the patients remaining >12 months in remission. When all patients evaluable were taken together, this frequency of malignant cells correlated strongly with disease-free survival (P < 0.001). Both this massive mobilization of normal CD34+ cells and high frequency of malignant cells in the subgroup of patients with CD34 >0.8% and relapse within 12 months indicate an insufficient in vivo purging, as well as low chemotherapeutic bone marrow toxicity. This was confirmed by an inverse correlation between hypoplasia period after the induction therapy and CD34% in PBSC products (P < 0.002). It is concluded that a subgroup of patients has been identified that might benefit from a more intensive chemotherapeutic treatment. PMID- 12529663 TI - Acute myeloid leukemia with MLL rearrangements: clinicobiological features, prognostic impact and value of flow cytometry in the detection of residual leukemic cells. AB - The MLL gene, located at 11q23 band, is frequently disrupted by different chromosomal rearrangements that occur in a variety of hematological malignancies. MLL rearrangements are associated with distinct clinical features and a poor prognosis. The aim of this study was to analyze the incidence and the prognostic significance of MLL rearrangements in a consecutive series of adult AML patients and to determine the immunophenotypic features of these cases. The identification of abnormal immunophenotypes could be used for the detection of minimal residual disease (MRD). Ninety-three adult patients with de novo acute myeloid leukemia (AML) were analyzed by Southern blot in order to detect MLL rearrangements (MLL+). RT-PCR and genomic long-range PCR were performed to further characterize MLL partial tandem duplication (PTD) in those patients in whom conventional karyotype did not show 11q23 chromosomal translocations. All the patients were homogeneously immunophenotyped at diagnosis. MLL rearrangements were detected in 13 (14%) patients. Four patients (5%) showed 11q23 translocations by karyotypic conventional analysis. Nine patients (10%) revealed PTD of MLL and one patient showed a MLL cleavage pattern. The MLL+ patients usually expressed myeloid and monocytic antigens CD33 (12/13 cases), CD13 (9/13), CD117 (9/13), CD64 (11/13) and in some cases CD14 (4/11). HLA-DR was also positive in (12/13). Eight out of 13 cases expressed the stem cell marker CD34. Only one patient revealed lymphoid marker reactivity (CD7) and CD56 was expressed in 5/13 cases. All the MLL+ patients showed at least one aberrant phenotype at diagnosis, which allowed us to set out a simple panel for the MRD studies. Twenty-seven samples from eight patients in morphologic complete remission (CR) were analyzed using the aberrant immunologic combinations detected at diagnosis. Phenotypically abnormal cells were detected in all the patients who subsequently relapsed, whereas only one patient with MRD+ remained in CR. Owing to the high level of residual leukemic cells, the MLL+ patients showed a short CR duration and a poor survival. In conclusion, immunophenotyping may be a suitable approach to investigating MRD status in AML patients with PTD of the MLL gene. PMID- 12529664 TI - Methylation of the hMLH1 promoter and its association with microsatellite instability in acute myeloid leukemia. AB - The hMLH1 and hMSH2 genes are involved in the DNA mismatch repair (MMR) pathway. Defects in either of these genes have been associated with genetic instability in a wide variety of malignancies. A molecular mechanism involved in aberrant MMR gene expression is the epigenetic silencing of transcription by promoter methylation. The importance of MMR promoter methylation in leukemia is presently unclear and we have therefore undertaken a detailed analysis of the promoter regions of hMLH1 and hMSH2 using the technique of bisulfite genomic sequencing. DNA from 55 patients with acute myeloid leukemia (AML) including 23 patients with therapy-related AML (t-AML) have been analyzed. Two patients with de novo AML demonstrated extensive methylation throughout the whole hMLH1 region sequenced, one of whom had previously shown widespread genetic instability, measured as microsatellite instability (MSI). However methylation of hMLH1 was not found in t AML which has previously been associated with MSI. In addition, methylation was seen at a restricted region of the hMLH1 promoter in both AML patients and healthy controls. The significance of this methylated region of the hMLH1 promoter is uncertain, however, our results confirm that in some patients with AML extensive methylation of hMLH1, but not of hMSH2 may occur, and as is the case in solid tumors this can be associated with the presence of a defective DNA mismatch repair pathway resulting in MSI. PMID- 12529665 TI - Clinicopathologic correlations of bone marrow angiogenesis in chronic myeloid leukemia: a morphometric study. AB - Various morphometric characteristics of microvessels, highlighted by means of anti-CD34 immunohistochemical staining, were evaluated in the bone marrow of 52 patients with chronic myeloid leukemia (CML) in chronic phase, in relation to several clinicopathologic parameters. Twenty control bone marrows and 15 cases of CML in blastic phase were also studied. Microvessel density (MVD), total vascular area (TVA) and several size- and shape-related parameters were quantitated in the region of most intense vascularization using image analysis. Overall, the group of chronic phase CML had higher MVD and size-related parameters and more branching microvessels than controls. Blastic phase was characterized by increased numbers of microvessels with a rounder shape and smaller caliber than chronic phase. A positive correlation emerged between marrow fibrosis and MVD as well as between white blood cell counts and rounder vessel sections. No relationship existed between microvascular parameters and Hasford or Sokal prognostic scores. In univariate analysis, overall and progression-free survival were adversely affected by MVD, size-related parameters, increased platelet count, age and spleen size. Multivariate analysis indicated that microvessel area was related to progression-free survival, whereas both MVD and area were significant prognosticators of overall survival, even when Hasford or Sokal scores are introduced into the model. Our data suggest that changes in angiogenic parameters may participate in the conversion of normal marrow to CML and ultimately to blastic transformation. More importantly, MVD and microvessel caliber are significant predictors of patient survival and progression. PMID- 12529666 TI - C-fms expression correlates with monocytic differentiation in PML-RAR alpha+ acute promyelocytic leukemia. AB - We have investigated the expression of the M-CSF receptor (c-fms) in 16 freshly isolated acute promyelocytic leukemias (APL) expressing the PML/RAR alpha fusion protein. In parallel, we evaluated the capacity of these cells to differentiate along the granulocytic and monocytic pathways. c-fms was constitutively and constantly expressed in all cases sensitive in vivo to all-trans retinoic acid (ATRA) and its expression was further potentiated following in vitro induction with ATRA. Furthermore, gel-shift analysis of APL cells showed elevated levels of PU.1 binding activity to the M-CSF receptor promoter, particularly after ATRA stimulation. Interestingly, the rise of PU.1 binding activity as well as of PU.1 levels after ATRA treatment was significantly higher in APL patients exhibiting monocytic maturation, as compared to those that did not undergo monocytic differentiation. A variable proportion of ATRA-induced APL cells exhibited monocyte-like morphology and immunophenotype: the proportion of monocytic cells was consistently increased by combined treatment with ATRA and diverse hematopoietic growth factors cocktails, which always comprised M-CSF. Monocytic cells originating from in vitro ATRA-induced maturation of APL cells derive from the leukemic clone as suggested by two lines of evidence: (1) monocytic cells harbor the 15;17 translocation; (2) monocytic cells possess Auer bodies. The c fms(bright) leukemic blasts preferentially showed the capacity for monocytic differentiation as compared to the c-fms(dim/-) subset: indeed, enforced expression of c-fms into NB4, a PML/RAR alpha+ cell line, favored the onset of monocytic maturation. Finally, low c-fms expression was observed in an APL relapsing patient resistant to ATRA, as well as in an APL case with t(11;17), PLZF/RAR alpha+. These observations indicate that PML/RAR alpha+ APL blasts are bipotent for differentiation through both neutrophilic and monocytic lineages, whereby monocytic differentiation is linked to c-fms expression and stimulation. PMID- 12529667 TI - Better detection of FLT3 internal tandem duplication using peripheral blood plasma DNA. AB - Somatic mutation of the FLT3 gene as an internal tandem duplication (ITD) of the juxtamembrane domain-coding sequence causes constitutive tyrosine phosphorylation and activation. Tumor-specific DNA has been documented in the sera of patients with solid tumors even when it is in an early stage. We compared the detection of FLT3 ITD in DNA extracted from cells of bone marrow (BM) aspirations with DNA extracted from peripheral blood (PB) plasma in patients newly diagnosed with acute myeloid leukemia (AML; 85 patients), myelodysplastic syndrome (MDS; 16 patients), and acute lymphocytic leukemia (ALL; 16 patients). FLT3 ITD was detected in 18 (21%) AML samples and in one (6%) MDS sample in both cellular and plasma DNA but in none of the ALL samples. Hemizygous/homozygous FLT3 ITD was detected in five (28%) of the FLT3 ITD-positive AML using plasma DNA, whereas only four of these cases showed hemizygous/homozygous FLT3 ITD using cellular DNA. The presence of FLT3 ITD was associated with significantly shorter survival (P = 0.02) when only patients younger than 50 years of age (48 AML+MDS patients) were considered. This finding was independent of cytogenetics in this age group. However, patients with the FLT3 ITD hemizygous/homozygous phenotype had even shorter survival (P = <0.001). As expected, the presence of FLT3 ITD correlated with higher white blood cell (WBC) counts. These data demonstrate that plasma DNA is a reliable alternative resource for detecting FLT3ITD, especially the hemizygous/homozygous genotype. Furthermore, the data derived from this study support the notion that the presence of FLT3 ITD in conjunction with the absence of the wild-type FLT3 allele predicts an especially poor prognosis for patients with AML. PMID- 12529668 TI - FLT3 mutations in acute myeloid leukemia cell lines. AB - Internal tandem duplications (ITD) and D835 point mutations of the receptor tyrosine kinase (RTK) FLT3 are found in a high proportion of cases with acute myeloid leukemia (AML). These genetic aberrations may lead to the constitutive activation of the receptor, thus providing the molecular basis for a persisting growth stimulus. We have screened 69 AML-derived cell lines for FLT3 mutations. Four of these cell lines showed ITD of the FLT3 gene, none carried a D835 point mutation. Two cell lines (MUTZ-11 and MV4-11) expressed exclusively the mutated allele, the other two cell lines (MOLM-13 and PL-21) displayed a mutated and the wild-type version of the gene. Although mutationally activated FLT3 is supposed to substitute for the stimulatory signal of a growth factor, one of these cell lines (MUTZ-11) was strictly cytokine-dependent. FLT3 transcripts were found in all four cell lines, but the constitutively phosphorylated receptor protein was clearly detectable only in cell line MV4-11, possibly explaining why MUTZ-11 cells were growth-factor dependent. Thus, not all FLT3 ITD-positive cells express high levels of the active receptor protein, a finding that might be of relevance for a possible future application of a kinase inhibitor as therapeutic agent. It had been described that STAT-5 phosphorylation was part of the FLT3 signalling chain and that STAT-5 molecules were constitutively phosphorylated in FLT3 ITD positive cells. Although we observed the constitutive phosphorylation of STAT-5 molecules in FLT3-mutant cells, FLT3 ligand (FL) did not induce STAT-5 phosphorylation in FLT3 wild-type cells. These results suggest that the signalling mechanisms of the mutated FL receptor differ at least to some extent from those conferred by wild-type FLT3. In conclusion, (1) not all cells with FLT3 ITD express significant amounts of the mutated receptor protein; (2) signals downstream from wild-type and mutant FLT3 receptors are not 100% identical; and (3) MV4-11 represents a model cell line for FLT3 ITD signalling. PMID- 12529669 TI - Diagnostic role and prognostic significance of a simplified immunophenotypic classification of mature B cell chronic lymphoid leukemias. AB - We verified the diagnostic and prognostic role of a simplified immunophenotypic classification (IC) in a series of 258 patients (M/F: 1.4; median age: 64 years; median follow-up: 64 months; 75 deaths) with mature B cell lymphoid leukemias (MBC-LL) for whom no histopathological diagnosis was available because of minimal or no lymph node involvement. The IC was based on the reactivity of three pivotal immunophenotypic markers: CD5, CD23 and SIg intensity. On the basis of different expression patterns, we identified four diagnostic clusters (C) characterized by distinct clinico-biological features and different prognoses: C1 (149 patients) identified most classical B cell chronic lymphocytic leukemias (CLL-type cluster; SIg(dim)/CD5+/CD23+); C2, 38 patients whose clinico-hematological characteristics were intermediate between C1 and C3 (CLL-variant cluster; SIg(bright)/CD5+/CD23+/ or SIg(dim)/CD5-/-/CD23 indifferent); C3 (16 patients) most situations consistent with mantle cell lymphoma in leukemic phase (MCL-type cluster; SIg(bright)/CD5+/CD23-); and C4, 55 cases, most of whom were consistent with leukemic phase lymphoplasmacytic/splenic marginal zone lymphomas (LP/S-type cluster; SIg(bright)/CD5-/+/CD23 indifferent). At univariate survival analysis, prognosis worsened from C1 to C4, C2 and C3 (P = 0.0001), and this was maintained at multivariate analysis (P = 0.006), together with CD11c expression (P = 0.0043), age at diagnosis (cut-off 70 years; P = 0.0008) and platelet count (cut off 140 x 10(9)/l; P = 0.0034). Besides recognising the two well-known situations of classic B-CLL and MCL, our IC identified situations with distinct prognostic and/or clinical behaviors. PMID- 12529670 TI - Predictive value of serum thymidine kinase level for Ig-V mutational status in B CLL. AB - In B-CLL IgV(H) genes mutational status is a major prognostic factor. Since sequencing of IgV(H) genes is not available in most laboratories, an easily performed surrogate assay is desirable. To identify the best surrogate assay, and to better discriminate prognostic subgroups we analyzed clinical and biological data from 58 typical CLL cases. A higher serum thymidine kinase level (>15 U/l) proved to be a strong predictor of mutational status, and the only independent one among the studied parameters. To further identify prognostic subgroups, cluster analysis was employed on 38 cases on which all data were available, which segregated two groups including 25 and 13 patients, respectively. These two clusters differed by their proliferative potential and appeared to discriminate patients with very different clinical course and outcome. s-TK was strikingly different among these two clusters, suggesting that s-TK level could be used routinely to identify patients at risk of progression. PMID- 12529671 TI - Flow cytometric follow-up of minimal residual disease in bone marrow gives prognostic information in children with acute lymphoblastic leukemia. AB - Using flow cytometry (FC) and live gate (LG) analysis we have followed levels of minimal residual disease (MRD) in the bone marrow (BM) of 70 consecutive patients with childhood acute lymphoblastic leukemia (59 B precursor ALL and 11 T-ALL) treated according to the Nordic (NOPHO-92) protocols. Thorough studies of B and T cell antigen expression patterns in normal BM performed during BIOMED 1 Concerted Action on MRD, made it possible to tailor individual protocols of marker combinations for follow-up in 97% of patients. In 12% of LG analyses, the numbers of cells exceeded 10(6) and in 82% exceeded 10(5), giving the sensitivity level of MRD detection 10(-5) and 10(-4), respectively. The median follow-up time was 53 months. Patients with MRD levels > or = 0.01% at follow-up time-points during and after first induction, and at the end of treatment had significantly lower disease-free survival by comparison to patients with MRD values <0.01%. Seven of nine patient with recurrence in the BM showed under treatment persisting MRD levels > or = 0.01% of BM cells. This was also observed in another two patients with infant leukemia who relapsed. In conclusion, the investigation of levels and the dynamics of MRD by sensitive and quantitative FC can provide a basis for further clinical studies for at least upgrading of therapy. PMID- 12529672 TI - Long-term study of the clinical significance of loss of heterozygosity in childhood acute lymphoblastic leukemia. AB - Acute lymphoblastic leukemia (ALL) is one of the most common malignancies in childhood, with a widely variable outcome. Differences in the behavior and prognosis of the leukemia suggest that ALL can be divided into several biologic subgroups. We analyzed the loss of heterozygosity (LOH) of 6q, 9p, 11q and 12p using 31 microsatellite sites to determine their overall frequency and clinical value. We have studied 244 primary ALL samples obtained from the Multicenter Trial ALL-BFM 90 of Childhood ALL group. These patients have now been followed clinically for over 8 years. LOH occurred in 169 (69%) individuals in the following frequencies: 6q, 49 patients (20%); 9p, 97 patients (40%); 11q, 29 patients (12%); 12p, 60 patients (25%). Clinical data showed that those with 6q LOH were younger (P = 0.01) and had lower WBC counts (P = 0.02); patients with 9p LOH more frequently had CNS involvement (P = 0.01) and T cell phenotype (P = 0.0001); individuals with 11q LOH had a good response to induction chemotherapy (P = 0.02); those with 12p LOH were younger (P = 0.005), frequently had precursor B ALL (P = 0.001), and had a longer event-free survival (P = 0.05). Taken together, these data confirm that LOH is a very frequent alteration in childhood ALL. PMID- 12529673 TI - Diphtheria toxin-interleukin-3 fusion protein (DT(388)IL3) prolongs disease-free survival of leukemic immunocompromised mice. AB - The novel fusion protein DT(388)IL3, composed of the catalytic and translocation domains of diphtheria toxin (DT(388)) fused with a Met-His linker to human interleukin 3 (IL-3), was tested for anti-leukemia efficacy in an in vivo model of differentiated human acute myeloid leukemia (AML). Six-week-old female SCID mice were irradiated with 350 cGy, inoculated 24 h later with 20 million (i.v., i.p., or s.c.) TF1 cells transfected with the v-SRC oncogene, and treated i.p., starting 24 h later, with up to five daily injections of saline, DT(388)IL3 (2 microg), DT(388)GMCSF (2 microg), DAB(389)IL2 (2 microg), or cytarabine (80 microg) or two weekly injections of anti-CD33-calicheamicin conjugate (5 microg). Animals were monitored twice daily, and moribund animals killed and necropsied. Control animals had a median disease-free survival (DFS) of 37 days (i.v., n = 45), 35 days (i.p., n = 20), and 21 days (s.c., n = 20), respectively. Only 5/49 (10%) of the DT(388)IL3 treated i.v. inoculated animals died with leukemia. Median DFS with i.v., i.p. and s.c. tumor inoculated animals was prolonged by fusion protein treatment to >120 days, 66 days and 31 days (P < 0.001, = 0.0003, and = 0.0006), respectively. Median DFS with s.c. tumor inoculated animals was also prolonged by other active anti-leukemia agents (DT(388)GMCSF, cytarabine and anti-CD33-calicheamicin) relative to controls by 67%, 172% and 47% (P < 0.0001, <0.0001, and =0.0004), respectively. In contrast, median DFS with s.c. tumor inoculated animals treated with DAB(389)IL2 non-significantly reduced by 13% relative to controls (P = 0.21). Thus, DT(388)IL3 fusion protein demonstrates in vivo anti-leukemia efficacy and warrants further preclinical development for treatment of chemo-resistant, IL-3 receptor positive AML patients. PMID- 12529674 TI - Primary murine MSC show highly efficient homing to the bone marrow but lose homing ability following culture. AB - Recent studies describe beneficial effects of bone marrow-derived mesenchymal stem cell infusion in animal models as well as in patients. However, data on the homing abilities of primary and culture-expanded MSC are lacking. In order to systematically investigate MSC homing we compared the fate of both primary and cultured MSC in a syngeneic mouse model. Twenty-four hours after transplantation of uncultured EGFP-transgenic MSC into sublethally irradiated mice, as many as 55 65% of injected CFU-F were recovered from the BM and 3.5-7% from the spleen. In the subsequent 4 weeks these donor CFU-F expanded 100-fold, which resulted in a normalization of femoral and splenic CFU-F numbers. This highly efficient homing of primary CFU-F contrasted with the defective homing of MSC following culture. Following their infusion immortalized multipotent syngeneic stromal cells were undetectable in BM, spleen, lymph nodes or thymus. Remarkably, following transplantation of primary MSC that had been cultured for only 24 h the seeding fraction in the BM was reduced to 10%, while after transplantation of 48 h cultured primary MSC no CFU-F were detected in the lymphohematopoietic organs. These data suggest that in vitro propagation of BM-derived MSC dramatically decreases their homing to BM and spleen. PMID- 12529675 TI - CD38 expression by hematopoietic stem cells of newborn and juvenile mice. AB - While investigators in a number of laboratories have documented that the hematopoietic stem cells (HSCs) of fetal and adult mice are CD38+, no information is available about CD38 expression by HSCs of newborn and juvenile mice. We used a murine transplantation model to examine HSC CD38 expression. First, we observed that all HSCs from newborn bone marrow are CD38-. Next, it was determined that the majority of HSCs in the bone marrow of 5-week-old mice are CD38-, with a minority being CD38+. These observations indicated that the CD38+ subpopulation of HSC appears before the age of 5 weeks and expands during adolescence. However, the majority of HSCs of 5-week-old mice became CD38+ following injection of 5 fluorouracil, indicating that activation of juvenile stem cells enhances CD38 expression. These observations may have implications for CD38 expression by HSCs from human umbilical cord blood and bone marrow of young children in steady state and under pathological conditions. PMID- 12529676 TI - Proteoglycans on bone marrow endothelial cells bind and present SDF-1 towards hematopoietic progenitor cells. AB - Recognition events between hematopoietic progenitor cells (HPC) and bone marrow endothelial cells (BMEC) initiate homing of HPC to the bone marrow. The chemokine SDF-1 is present on BMEC and plays a crucial role in bone marrow engraftment. We studied the role of proteoglycans (PGs) on BMEC in binding and presentation of SDF-1. SDF-1 mRNA was present in three human BMEC cell lines. Competition experiments showed that 125I-SDF-1 alpha binding to the BMEC cell line 4LHBMEC was inhibited by heparins, heparan sulfate (HS) intestinal mucosa, chondroitin and dermatan sulfate (CS/DS), but not by HS bovine kidney. Pretreatment of 4LHBMEC with glycosaminoglycan (GAG)-degrading enzymes or sodium chlorate demonstrated that SDF-1 bound to both HSPGs and CS/DSPGs in a sulfation-dependent manner, as determined with an SDF-1 antibody recognizing the CXCR4-binding site. 4LHBMEC bound four-fold more SDF-1 than HUVEC. Isolated endothelial PGs did not bind SDF-1 in a filter or microplate-binding assay, suggesting the necessity of membrane association. In flow adhesion experiments, endothelial arrest of CXCR4+ KG-1 and not of CXCR4- KG-1a cells increased significantly when SDF-1 was presented on 4LHBMEC. In conclusion, SDF-1 is produced by BMEC and binds to the BMEC cell surface via HS and CS/DS-GAGs, thereby presenting its CXCR4 binding site to HPC contributing to their arrest. PMID- 12529677 TI - Qualitative and quantitative analysis of human herpesviruses in chronic and acute B cell lymphocytic leukemia and in multiple myeloma. AB - Real-time quantitative polymerase chain reaction (qPCR) was used to quantify viral loads of human herpesviruses (HHVs) at diagnosis in 61 samples of malignant B cells: 21 chronic lymphocytic leukemia (B-CLL), 29 acute lymphoblastic leukemia (B-ALL) and 11 multiple myeloma (MM); control samples were blasts from 16 acute myeloid leukemia (AML) and 24 blood or bone marrow samples from healthy donors. The majority of samples from healthy donors and patients (B-ALL, B-CLL or AML, but not MM) was positive for EBV and contained <100 ebv copies/100 ng dna. ebv loads were occasionally high (>500 copies/100 ng DNA) in B-ALL (2/16) and in B CLL (2/21) samples. The fractions of samples positive for HHV-8 and HHV-6A, less than 10% for MM patients, were high for adults with B-ALL (18.8% HHV-8+, 43.8% HHV-6A+) or B-CLL (28.6% HHV-8+, 52.4% HHV-6A+). B-ALL, B-CLL and MM samples were rarely positive for HHV-6B and HHV-7. Lastly, 75% of B-ALL samples were positive for CMV, and CMV loads were significantly higher in B-ALL samples than in MM, B CLL or AML samples. We also used PCR with consensus-degenerate hybrid oligonucleotide primers (CODEHOP) to look for novel HHVs in B cell samples: no sequence indicative of a new HHV was detected. Altogether, the data indicate that the presence of multiple HHVs, including EBV and CMV at high loads, is not rare in B-ALL and B-CLL cell samples. Future prospective studies should determine whether patients with high EBV/CMV loads at diagnosis in tumor samples face a higher risk of delayed hematological recovery, virus-related complications or relapse. PMID- 12529678 TI - Clinical significance of nm23-H1 proteins expressed on cell surface in non Hodgkin's lymphoma. AB - The nm23 gene was isolated as a metastasis suppressor gene that exhibits low expression in high-level metastatic cancer cells. Its gene is related to the prognosis of acute myelogenous leukemia (AML) and non-Hodgkin's lymphoma (NHL). In this study, we examined the expression of nm23-H1 protein on the lymphoma cell surface of NHL. In 28 of 108 cases (25.9%), we observed > or = 20% of cell surface nm23-H1 protein expression and expression was especially high in peripheral T cell lymphomas and extranodal NK/T cell lymphomas. We also observed a significant correlation between serum nm23-H1 level and cell surface nm23-H1 expression levels. In patients with high levels of cell surface nm23-H1 expression, overall and progression-free survival rates were significantly lower than those in patients with low surface nm23-H1 expression levels. When surface nm23-H1 and serum nm23-H1 were combined, patients with high levels of both exhibited a poorer prognosis than patients with a high level of one or the other. These results indicate that in addition to serum nm23-H1, cell surface nm23-H1 may be used as a prognostic factor in planning a treatment strategy. The nm23-H1 protein appears to be intimately related to biological aggressiveness of lymphoma and, therefore, might be a molecular target of NHL treatment. PMID- 12529679 TI - Expression and function of chemokine receptors in human multiple myeloma. AB - Multiple myeloma (MM) is a B cell tumor characterized by its selective localization in the bone marrow. The mechanisms that contribute to the multiple myeloma cell recruitment to the bone marrow microenvironment are not well understood. Chemokines play a central role for lymphocyte trafficking and homing. In this study we have investigated expression and functional importance of chemokine receptors in MM-derived cell lines and primary MM cells. We found that MM cell lines express functional CCR1, CXCR3 and CXCR4 receptors, and some also CCR6. Although only a minority of the cell lines responded by calcium mobilization after agonist stimulation, a migratory response to the CCR1 ligands RANTES and MIP-1 alpha was obtained in 5/6 and 4/6, respectively, of the cell lines tested. Five out of six cell lines showed a response to the CXCR4 ligand SDF-1. In addition, 3/6 cell lines migrated in response to MIP-3 alpha and IP-10, ligands for CCR6 and CXCR3, respectively. The expression of CXCR4 and CCR1 and the migration to their ligands, SDF-1, and RANTES and MIP-1 alpha, respectively, were also demonstrated in primary MM cells. These findings suggest that chemokine receptor expression and the migratory capacity of MM cells to their ligands are relevant for the compartmentalization of MM cells in the bone marrow. PMID- 12529682 TI - Interference of dye-associated fluorescence signals with quantitative analysis of chimerism by capillary electrophoresis. PMID- 12529680 TI - Chemosensitization of myeloma plasma cells by an antisense-mediated downregulation of Bcl-2 protein. AB - An antisense oligodeoxynucleotide (ODN) complementary to the first six codons of the Bcl-2 mRNA, G3139 (oblimersen sodium; Genasense), has been shown to downregulate Bcl-2 and produce responses in a variety of malignancies including drug-resistant lymphoma. Incubation of ex vivo purified plasma cells from patients with multiple myeloma (MM) with carboxyfluorescein (FAM)-labeled antisense ODNs resulted in a time- and dose-dependent uptake in the cytoplasm and nucleus. No major differences in uptake of Bcl-2 antisense ODNs were observed among patients' samples. Incubation of purified myeloma plasma cells with G3139, but not solvent or reverse polarity control ODNs, resulted in a reduction (>75%) of Bcl-2 mRNA levels after 2 and 4 days, as measured by Real-Time PCR. Treatment with G3139 led to a sequence-specific reduction of Bcl-2 protein levels within 4 days of exposure in 10 out of 11 clinical samples from patients with chemosensitive and multidrug-resistant disease, without significant reduction of alpha-Actin, Bax, Bcl-XL, or Mcl-1 proteins. This resulted in a significantly enhanced sensitivity of the myeloma tumor cells to dexamethasone or doxorubicin induced apoptosis. G3139 can consistently enter myeloma cells, downregulate the expression of Bcl-2, and enhance the efficacy of myeloma therapy. These data support further clinical evaluation of G3139 therapy in multiple myeloma. PMID- 12529683 TI - Quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection: the Vienna experience. PMID- 12529684 TI - Quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection: the Geneva experience. PMID- 12529685 TI - Quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection: the Frankfurt experience. PMID- 12529686 TI - Quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection: the Tuebingen experience. PMID- 12529687 TI - Quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection: the Paris-Robert Debre experience. PMID- 12529688 TI - Quantitative analysis of chimerism after allogeneic bone marrow transplantation using immunomagnetic selection and fluorescent microsatellite PCR. PMID- 12529689 TI - Summary: reports on quantitative analysis of chimerism after allogeneic stem cell transplantation by PCR amplification of microsatellite markers and capillary electrophoresis with fluorescence detection. PMID- 12529690 TI - Is t(14;18)(q32;q21) a constant finding in follicular lymphoma? An interphase FISH study on 63 patients. AB - The translocation (14;18)(q32;q21) is the hallmark of follicular lymphoma (FL). However, conventional cytogenetics and PCR techniques fail to detect it in at least 10% of cases. In order to evaluate the true incidence of this translocation in FL, we analyzed 63 patients with FL, and 17 patients with diffuse large cell lymphoma (DLCL) corresponding to suspected FL transformations using interphase fluorescence in situ hybridization (FISH). Colocalized signals related to the translocation were observed in 19-92% of cells (median = 51%), corresponding to positivity over the threshold in all (63/63) cases. Similarly, 16/17 possibly secondary DLCL displayed the translocation. Although some cytogenetic changes might be missed by this FISH assay (such as rare insertion, or translocations with other chromosomal partners), our results stress t(14;18)(q32;q21) as an almost constant finding in FL. Our sensitive interphase FISH assay should be of great value to define FL more accurately, namely in patients included into therapeutic trials. Furthermore, this approach could be of interest in (re)defining some types of FL, especially the grade 3 FL which frequently lack t(14;18). PMID- 12529691 TI - IMGT databases, web resources and tools for immunoglobulin and T cell receptor sequence analysis, http://imgt.cines.fr. AB - IMGT, the international ImMunoGeneTics database((R)) (http://imgt.cines.fr), is a high-quality integrated information system specializing in immunoglobulins (IG), T cell receptors (TR) and major histocompatibility complex (MHC) of human and other vertebrates, created in 1989, by LIGM, at the Universite Montpellier II, CNRS, Montpellier, France. IMGT provides a common access to standardized data which include nucleotide and protein sequences, oligonucleotide primers, gene maps, genetic polymorphisms, specificities, 2D and 3D structures. IMGT includes several databases (IMGT/LIGM-DB, IMGT/3Dstructure-DB, IMGT/HLA-DB), Web resources ('IMGT Marie-Paule page') and interactive tools (IMGT/V-QUEST, IMGT/JunctionAnalysis). IMGT expertly annotated data and tools described in this paper are particularly useful for the analysis of the IG and TR rearrangements in leukemia, lymphoma and myeloma, and in translocations involving the antigen receptor loci. IMGT is freely available at http://imgt.cines.fr. PMID- 12529692 TI - Ubiquinone does not rescue acute myeloid leukemia cells from growth inhibition by statins. PMID- 12529693 TI - Inhibition affecting RQ-PCR-based assessment of minimal residual disease in acute lymphoblastic leukemia: reversal by addition of bovine serum albumin. PMID- 12529694 TI - Clinical activity of arsenic trioxide in Burkitt-like lymphoma. PMID- 12529696 TI - Mutation of BRAF is uncommon in AML FAB type M1 and M2. PMID- 12529695 TI - Zinc modulates c-Myc/Mad1 balance in human leukemia cells. PMID- 12529697 TI - Genetic discrimination: too few data. PMID- 12529698 TI - Discovery of old diseases: the molecular approach. PMID- 12529699 TI - Absence of association of fetal MTHFR C677T polymorphism with prenatal Down syndrome pregnancies. PMID- 12529701 TI - Linkage of aggressive prostate cancer to chromosome 7q31-33 in German prostate cancer families. AB - It has been suggested that chromosome 7q32 contains genes that influence the progression of prostate cancer from latent to invasive disease. In an attempt to confirm this linkage to prostate cancer aggressiveness, 100 German prostate cancer families were genotyped using a panel of eight polymorphic markers on chromosome 7q. We used a multipoint allele sharing method based upon a likelihood ratio test implemented in GENEHUNTERPLUS v1.2 in order to calculate the nonparametric Z(lr) and the associated LOD scores. We applied the aggressiveness of prostate cancer given by the pathological tumour grade of each individual, and the mean age of onset of a family as covariates, and constructed two weighted models. The first (weight(0-1) model) puts weights on families with at least two cases of GIII prostate cancer. The second (weight(0-2) model) also adds weights to families with early and late onset cancer respectively. The unweighted analysis gave no evidence of linkage to chromosome 7q. The Z(lr) scores increased when including the covariates, to 2.60 (P=0.005) using the weight(0-1) and to 3.02 (P=0.001) using the weight(0-2) model for late onset prostate cancer. The associated LOD scores were respectively 1.47 (P=0.009) and 1.98 (P=0.002). The markers that gave most evidence for linkage were exactly in the range of the published prostate cancer aggressiveness region. Our results support a widespread relevance of this locus and suggest that aggressive and late onset prostate cancer is linked to chromosme 7q31-33 in the German population. PMID- 12529700 TI - Haplotype structure and association to Crohn's disease of CARD15 mutations in two ethnically divergent populations. AB - Current debate focuses on the relevance of linkage disequilibrium (LD), ethnicity and underlying haplotype structure to the search for genes involved in complex disorders. The recently described association between single nucleotide polymorphisms (SNPs) of the CARD15 (NOD2) gene and Crohn's disease (CD) in populations of north-European descent provides a test case that we have subjected to detailed SNP haplotype based analyses. We examined 23 SNPs spanning 290 kb, including CARD15, in large North-European and Korean samples of patients with Crohn's disease and normal controls. In Europeans we confirmed that the three disease-associated SNPs occur independently but share a common background haplotype. This suggests a common origin and the possibility of an undiscovered more strongly predisposing mutation. Korean CD patients present a phenotype identical to the European patients and have not previously been screened for CARD15. The three disease-associated SNPs were absent and there was no evidence of association between CARD15 and CD. Consequently, the disease-associated mutations in the Europeans, which are rare, have arisen recently (after the Asian European split). Our results highlight important issues relevant to mapping the genes that predispose to complex disorders. First, although ethnically divergent populations may present identical phenotypes they do not necessarily share the same set of predisposing genes. Second, although single-locus tests of association showed consistent association with markers throughout the gene, pair wise LD between markers (r(2) and D') yielded very little information about actual disease-association. Third, a population comparative approach allowed refining of the marker set through the examination of shared polymorphisms and common LD-groups. This approach, in conjunction with the examination of the mutational steps in a haplotype network, allows unambiguous identification of the potentially causative mutations. PMID- 12529702 TI - Cystathionine beta-synthase polymorphisms and hyperhomocysteinaemia: an association study. AB - Hyperhomocysteinaemia is generally accepted as an independent and graded risk factor for both arterial occlusive disease and venous thrombosis. The only way of homocysteine degradation is conversion to cysteine in the transsulfuration pathway in which the regulating step is catalysed by cystathionine beta-synthase (CBS). Mild impairment of CBS function could therefore affect homocysteine concentration, in particular after methionine loading, and consequently cardiovascular disease (CVD) risk. We analysed two silent polymorphisms and one short tandem repeat in the CBS gene (ie 699C-->T, 1080C-->T and -5697 (GT) STR) as genetic markers potentially in linkage disequilibrium with a functional polymorphism. We assessed their association with fasting and post-methionine load homocysteine in 190 patients with arterial occlusive disease, and in 381 controls. No differences in CBS genotype frequencies between cases and controls were found, nor was a particular CBS genotype associated with an elevated risk of arterial occlusive disease. Although we did find a high rate of linkage disequilibrium between the two single nucleotide polymorphisms and the GT STR, none of the genotypes defined by the three CBS variants studied showed an association with elevated fasting, post-load or increase upon methionine loading homocysteine concentrations. In conclusion, we did not find any indication that genetic variation in the CBS gene is associated with increased homocysteine concentrations. PMID- 12529703 TI - Psychological distress in the 5-year period after predictive testing for Huntington's disease. AB - The paper reports on a 5-year longitudinal study on psychological distress after predictive testing for Huntington's disease (HD) and on correlates of post-test distress. Psychometric tests and questionnaires were used. The tested persons were invited to participate in the follow-up study; the uptake rate was 75% (24 carriers, 33 non-carriers). Three time points were included: baseline, 1 year and 5 years post-test. Five years after the test, mean distress scores of both carriers and non-carriers were within the normal range. Carriers did not differ from non-carriers with regard to mean general distress. Compared to non-carriers, however, carriers had significantly less positive feelings (P<0.001) and were more consciously avoiding HD-related situations and thoughts (P<0.01). These findings reflect the carriers' conscious and unconscious attempt to escape from pessimism and to minimise negative consequences of the test result. Psychological distress 5 years post-test was significantly associated with ego-strength (P<0.05 to P<0.001). Except for intrusion and avoidance, distress was also associated with test motivation (P<0.05 to P<0.01). Compared with baseline level, mean depression, general and specific anxiety had significantly decreased 1 year and 5 years post-test (P<0.05 to 0.01). This evolution was independent of the test result. However, based on test motivation, a subgroup of tested persons having long lasting psychological distress could be identified, also irrespective of test result. Persons who asked the test to get rid of the uncertainty, without being able to specify implications for substantial life areas, had more psychological distress before and after the test than those who wanted the test for specific reasons (P<0.001 to P<0.0001). Moreover, the pattern of post-test anxiety differed over time, depending on the test motivation (P<0.05). The findings suggest that pre- and post-test counselling should pay special attention to persons with lower ego-strength and with an unspecified test motivation, because they are at higher risk for long-term psychological distress, independently of the test result. PMID- 12529704 TI - Global genetic variation at nine short tandem repeat loci and implications on forensic genetics. AB - We have studied genetic variation at nine autosomal short tandem repeat loci in 20 globally distributed human populations defined by geographic and ethnic origins, viz., African, Caucasian, Asian, Native American and Oceanic. The purpose of this study is to evaluate the utility and applicability of these nine loci in forensic analysis in worldwide populations. The levels of genetic variation measured by number of alleles, allele size variance and heterozygosity are high in all populations irrespective of their effective sizes. Single- as well as multi-locus genotype frequencies are in conformity with the assumptions of Hardy-Weinberg equilibrium. Further, alleles across the entire set of nine loci are mutually independent in all populations. Gene diversity analysis shows that pooling of population data by major geographic groupings does not introduce substructure effects beyond the levels recommended by the National Research Council, validating the establishment of population databases based on major geographic and ethnic groupings. A network tree based on genetic distances further supports this assertion, in which populations of common ancestry cluster together. With respect to the power of discrimination and exclusion probabilities, even the relatively reduced levels of genetic variation at these nine STR loci in smaller and isolated populations provide an exclusionary power over 99%. However, in paternity testing with unknown genotype of the mother, the power of exclusion could fall below 80% in some isolated populations, and in such cases use of additional loci supplementing the battery of the nine loci is recommended. PMID- 12529705 TI - Familial Mediterranean fever is no longer a rare disease in Italy. AB - Familial Mediterranean fever (FMF) is an autosomal recessive disorder, characterised by short, recurrent attacks of fever with abdominal, chest or joint pain and erysipelas-like erythema. It is an ethnically restricted genetic disease, found commonly among Mediterranean populations, as well as Armenians, Turks, Arabs and Jews. Traditionally, Italians have been considered little affected by FMF, despite the geographical position of Italy (northern Mediterranean basin) and the migratory changes in its population. The objective was to characterise the demographic, clinical and genetic features of FMF in Italy. Patients of Italian origin were recruited from those referred to Italian French medical centres for FUO (Fever of Unknown Origin) or 'surgical' emergencies; clinical history, genealogy and physical examination were recorded; all other possible infectious, neoplastic, auto-immune and metabolic diseases were excluded. Mutational analysis of the gene responsible for FMF (MEFV on 16p13.3) was performed, after which geno-phenotypical correlations were established. Italian FMF patients, 40 women and 31 men, aged from 3 to 75 years, have shown all the clinical manifestations indicative of FMF described in the literature, but with a lower incidence of amyloidosis. The genetic tests have been contributive in 42% of cases. The frequency of each different mutation has been similar to that found in a series of 'endemic' countries. The geno phenotypical correlations have suggested the existence of genetic and/or environmental modifier-factors. Among Italians FMF seems to be more frequent than was believed in the past. The data presented are consistent with their geographical location and their history. PMID- 12529706 TI - A new web-based data mining tool for the identification of candidate genes for human genetic disorders. AB - To identify the gene underlying a human genetic disorder can be difficult and time-consuming. Typically, positional data delimit a chromosomal region that contains between 20 and 200 genes. The choice then lies between sequencing large numbers of genes, or setting priorities by combining positional data with available expression and phenotype data, contained in different internet databases. This process of examining positional candidates for possible functional clues may be performed in many different ways, depending on the investigator's knowledge and experience. Here, we report on a new tool called the GeneSeeker, which gathers and combines positional data and expression/phenotypic data in an automated way from nine different web-based databases. This results in a quick overview of interesting candidate genes in the region of interest. The GeneSeeker system is built in a modular fashion allowing for easy addition or removal of databases if required. Databases are searched directly through the web, which obviates the need for data warehousing. In order to evaluate the GeneSeeker tool, we analysed syndromes with known genesis. For each of 10 syndromes the GeneSeeker programme generated a shortlist that contained a significantly reduced number of candidate genes from the critical region, yet still contained the causative gene. On average, a list of 163 genes based on position alone was reduced to a more manageable list of 22 genes based on position and expression or phenotype information. We are currently expanding the tool by adding other databases. The GeneSeeker is available via the web-interface (http://www.cmbi.kun.nl/GeneSeeker/). PMID- 12529707 TI - PTPN11 mutations are not responsible for the Cardiofaciocutaneous (CFC) syndrome. AB - Cardiofaciocutaneous (CFC) syndrome is a multiple congenital anomalies/mental retardation syndrome characterized by congenital heart defects, characteristic facial appearance, short stature, ectodermal abnormalities and mental retardation. It was described in 1986, and to date is of unknown genetic etiology. All reported cases are sporadic, born to non-consanguineous parents and have apparently normal chromosomes. Noonan and Costello syndromes remain its main differential diagnosis. The recent finding of PTPN11 missense mutations in 45-50% of the Noonan patients studied with penetrance of almost 100% and the fact that in animals mutations of this gene cause defects of semilunar valvulogenesis, made PTPN11 mutation screening in CFC patients a matter of interest. We sequenced the entire coding region of the PTPN11 gene in ten well-characterised CFC patients and found no base changes. We also studied PTPN11 cDNA in our patients and demonstrated that there are no interstitial deletions either. The genetic cause of CFC syndrome remains unknown, and PTPN11 can be reasonably excluded as a candidate gene for the CFC syndrome, which we regard as molecular evidence that CFC and Noonan syndromes are distinct genetic entities. PMID- 12529708 TI - Arrhythmogenic right ventricular cardiomyopathy type 1 (ARVD1): confirmation of locus assignment and mutation screening of four candidate genes. AB - Arrhythmogenic right ventricular cardiomyopathy type 1 (ARVD1) is an autosomal dominant disorder characterised by progressive degeneration of right ventricular myocardium, arrhythmias and risk of sudden death. By linkage analysis, we previously mapped the involved gene to chromosome 14q24.3. In the present study we report on linkage analysis of one additional and unrelated family, which enabled to confirm previous locus assignment. Another family is reported, in which genetic and clinical data suggest linkage to the same locus. Direct sequencing of DNA from individuals belonging to established ARVD1 families failed to detect causative mutations in exonic sequences of four genes (POMT2, TGFbeta3, KIAAA1036 and KIAA0759) expressed in the heart and which defects could possibly induce plasma membrane instability or apoptosis, key features of ARVD pathogenesis. PMID- 12529709 TI - A novel autosomal recessive non-syndromic deafness locus (DFNB35) maps to 14q24.1 14q24.3 in large consanguineous kindred from Pakistan. AB - Autosomal recessive nonsyndromic deafness is one of the most frequent forms of inherited hearing impairment. Over 30 autosomal recessive nonsyndromic hearing loss loci have been mapped, and 15 genes have been isolated. Of the over 30 reported autosomal recessive nonsyndromic hearing loss (NSHL) loci, the typical phenotype is prelingual non-progressive severe to profound hearing loss with the exception of DFNB8, which displays postlingual onset and DFNB13, which is progressive. In this report we describe a large inbred kindred from a remote area of Pakistan, comprising six generations and segregating autosomal recessive nonsyndromic prelingual deafness. DNA samples from 24 individuals were used for genome wide screen and fine mapping. Linkage analysis indicates that in this family the NSHL locus, (DFNB35) maps to a 17.54 cM region on chromosome 14 flanked by markers D14S57 and D14S59. Examination of haplotypes reveals a region that is homozygous for 11.75 cM spanning between markers D14S588 and D14S59. A maximum two-point LOD score of 5.3 and multipoint LOD score of 7.6 was obtained at marker D14S53. The interval for DFNB35 does not overlap with the regions for DFNA9, DFNA23 or DFNB5. PMID- 12529710 TI - The CTLA4 region as a general autoimmunity factor: an extended pedigree provides evidence for synergy with the HLA locus in the etiology of type 1 diabetes mellitus, Hashimoto's thyroiditis and Graves' disease. AB - We have identified a large family in the northern part of Sweden with multiple cases of autoimmune diseases, namely type 1 diabetes (T1D), Graves' disease (GD) and Hashimoto's thyroiditis (HT). The family members affected by any of these diseases share a region of 2.4 Mb that comprises among others the CTLA4 gene. We determined that all affected members of the family shared the HLA susceptibility haplotype (DR4-DQA1*0301-DQB1*0302). Analysis of genetic interaction conditioning for HLA haplotype provided strong evidence that the critical region which includes the CTLA4 gene acts together with the HLA locus on the etiology of disease (lodscore 4.20 (theta=0.0). The study of this family allowed us to: (1) reinforce a number of reports on linkage and association of the CTLA4 region to T1D and AITD; (2) demonstrate that a single haplotypic variant in this region constitutes an etiological factor to disease susceptibility in T1D, GD and HT; (3) reveal a strong genetic interaction of the CTLA4 and HLA loci in the genetic architecture of autoimmune disease; (4) emphasise the value of large pedigrees drawn from isolated populations as tools to single out the effect of individual loci in the etiology of complex diseases. PMID- 12529711 TI - PTPN11 mutation in a large family with Noonan syndrome and dizygous twinning. AB - Noonan syndrome (NS, MIM 163950) is an autosomal dominant condition characterised by facial dysmorphy, congenital cardiac defects and short stature. Recently missense mutations in PTPN11, the gene encoding the nonreceptor protein tyrosine phosphatase SHP-2 on 12q24, were identified in 50% of analysed Noonan cases. A large four-generation Belgian family with NS and some features suggestive of cardio-facio-cutaneous syndrome (CFC) was previously used to fine map the Noonan syndrome candidate region to a 5 cM region in 12q24. We now report the identification of a mutation (Gln79Arg) in the PTPN11 gene in this large family. In D. melanogaster and C. elegans the PTPN11 gene has been implicated in oogenesis. In this family two affected females had dizygous twins. This suggests that PTPN11 might also be involved in oogenesis and twinning in humans. PMID- 12529712 TI - Cryptic subtelomeric 6p deletion in a girl with congenital malformations and severe language impairment. AB - Several cases with microscopically visible, terminal 6p deletions have been described, and a distinct clinical phenotype has emerged, including developmental delay, congenital heart malformations, ocular abnormalities, hearing loss and a characteristic facial appearance. We report a patient with a submicroscopic 6p deletion, detected by subtelomeric screening using fluorescence in situ hybridisation. This girl presented with typical facial dysmorphic features, hearing impairment, malformation of the anterior eye segment, an ASD and severe language impairment. However, her cognitive functions were within the normal range. Detailed FISH analysis with 20 BAC probes covering the distal 6p25 region estimated the size of the terminal deletion to 2.1 Mb, and thus this case narrows down the critical region for the 6p phenotype. The forkhead transcription factor gene FOXC1, involved in a spectrum of anterior eye chamber disorders, is deleted in this patient, together with several characterised and putative genes with yet unknown function. PMID- 12529713 TI - Cationic trypsinogen and pancreatic secretory trypsin inhibitor gene mutations in neonatal hypertrypsinaemia. AB - Neonatal hypertrypsinaemia with normal sweat chloride detected during CF screening may be related to trypsin activation. We have looked for mutations of the cationic trypsinogen (PRSS1) and pancreatic secretory trypsin inhibitor (PSTI) genes in 50 hypertrypsinaemic neonates with known CFTR genotypes and negative sweat test. No mutations were found in either gene. Two silent polymorphisms were detected in the PRSS1 gene. A polymorphism in the promoter region and an intronic polymorphism of the PSTI gene were found. No difference was observed in the frequency of PRSS1 or PSTI polymorphisms in neonates carrying or not carrying CF mutations. These results do not provide an indication for an increased frequency of mutations in the PRSS1 and PSTI genes in this group of neonates with transient hypertrypsinaemia. PMID- 12529714 TI - A novel mutation in the G4.5 (TAZ) gene in a kindred with Barth syndrome. AB - Barth syndrome is an X-linked recessive disorder characterised by dilated cardiomyopathy and a variable expression of skeletal myopathy, short statue and neutropenia. Molecular genetic analysis is currently the most reliable diagnostic method. A kindred with a novel 535delC mutation in the G4.5 (TAZ) gene responsible for Barth syndrome is presented. Beside the patient, the same mutation was detected in patient's mother and grandmother. In contrast to the so far reported patients with mutations in the same region of G4.5 (TAZ) gene, the patient described here has only a mild and transitory clinical presentation. This could be attributed to alternative splicing of G4.5 (TAZ) gene, since mRNA lacking exon 6 (with 535delC mutation) was detected. Genetic analysis of the G4.5 (TAZ) gene was helpful for establishing the precise diagnosis of Barth syndrome and for adequate genetic counselling. Predicting the phenotype on the basis of mutations is unreliable especially if mutations are localised in alternatively spliced exons of the G4.5 (TAZ) gene which may result in a milder clinical presentation than expected. PMID- 12529715 TI - Four novel thymidine phosphorylase gene mutations in mitochondrial neurogastrointestinal encephalomyopathy syndrome (MNGIE) patients. AB - Mitochondrial neurogastrointestinal encephalomyopathy syndrome (MNGIE) is a rare autosomal recessive neurologic disorder characterised by multiple mitochondrial DNA deletions. In this study, five Turkish MNGIE patients are investigated for mtDNA deletions and TP gene mutations. The probands presented all the clinical criteria of the typical MNGIE phenotype; the muscle biopsy specimens also confirmed the diagnosis with ragged red fibres and cytochrome C oxidase (COX) negative fibres. The mitochondrial DNA analysis revealed no deletions in the probands' skeletal muscle samples. We have identified four novel mutations in the TP gene while one of the patients also harboured a nucleotide change, which was previously reported as a mutation. PMID- 12529718 TI - Future changes in the GDS. PMID- 12529719 TI - Hygienists using lasers. PMID- 12529721 TI - Further statistics in dentistry. Part 6: Multiple linear regression. PMID- 12529722 TI - Ethics: How the Apothecaries Act of 1815 shaped the dental profession. Part 2. The chemist-dentists and the education of dentists. PMID- 12529723 TI - A view from the bodies corporate. 4. Boots Wellbeing Services. Interview by F. Stuart-Wilson. PMID- 12529726 TI - The impact of attendance patterns on oral health in a general dental practice. AB - OBJECTIVE: The aim of this study was to examine the impact of attendance patterns on oral health in the context of government policy on dental care and registration in the UK. METHOD: The data involved 643 consecutive patient responses to a questionnaire on dental health taken from a survey that was conducted during 1998 in an urban area of Swansea. The survey continued for a period of six months and covered patients of 18 years of age and over, responding to a questionnaire on the subjective oral health status indicators including the pattern of their attendance to dental practices. RESULTS: Regular dental care attendance has a significant positive impact on dental health while its impact on the number of teeth present is insignificant. Regular attendees also suffer significantly less from the severity, prevalence, social and psychological impacts of dental health problems. CONCLUSION: Regular dental attendance is associated with better oral health when regularity of care is defined as a visit within a two-year period. The rationality of a 15 month registration period is therefore debatable in the context of developing equitable services. PMID- 12529727 TI - Assessing attitudes in dental education: is it worthwhile? PMID- 12529733 TI - Platelet glycoprotein IIb/IIIa inhibitors and thrombocytopenia: possible link between platelet activation, autoimmunity and thrombosis. PMID- 12529734 TI - Another interaction of the FcR system with IVIG. PMID- 12529735 TI - Acute thrombocytopenia in patients treated with the oral glycoprotein IIb/IIIa inhibitors xemilofiban and orbofiban: evidence for an immune etiology. AB - Thrombocytopenic episodes occurring in 18,845 patients treated with the GPIIb/IIIa inhibitors xemilofiban and orbofiban ("fibans") were analyzed by a blinded review panel and 73 patients were classified as having "possible fiban induced thrombocytopenia". When the treatment codes were broken, a significant association between drug exposure and assignment to this group was found (p <0.001). Twenty-eight (82%) of 34 archived serum samples from these patients contained fiban-dependent antibodies specific for GPIIb/IIIa, but no such antibodies were found in 61 drug treated patients not classified as having "possible fiban-induced thrombocytopenia" (p <0.001). We conclude that fiban dependent antibodies were the major cause of acute, severe thrombocytopenia in patients judged on the basis of clinical findings to have thrombocytopenia "possibly-induced" by xemilofiban and orbofiban. Measurement of drug-dependent antibodies may be helpful in determining the basis for acute thrombocytopenia in fiban-treated patients and possibly for identification of patients at risk to develop thrombocytopenia. PMID- 12529736 TI - Intravenous immunoglobulin mediates an increase in anti-platelet antibody clearance via the FcRn receptor. AB - We have recently shown that intravenous immunoglobulin (IVIG) therapy leads to an increased rate of anti-platelet antibody clearance in an animal model of immune thrombocytopenia. The present study was performed to confirm the importance of the FcRn receptor in mediating this effect of IVIG. The pharmacokinetics of an anti-platelet antibody, 7E3, were studied in mice lacking expression of FcRn and in control mice, both in the presence and absence of IVIG. IVIG increased the clearance of 7E3 in mice with functioning FcRn receptors, with an average clearance value of 14.4 +/- 1.4 ml/d/kg in IVIG treated mice vs. 5.2 +/- 0.3 ml/d/kg in controls (P <0.001). In mice lacking expression of FcRn, IVIG treatment did not increase 7E3 clearance (61.0 +/- 3.6 ml/d/kg vs. 71.5 +/- 4.0 ml/d/kg in controls). Thus, these data support the hypothesis that IVIG increases antibody elimination via saturation of FcRn. PMID- 12529737 TI - Thrombosis in children: current perspective and distinct challenges. AB - Coincident with a true increase in the incidence of thrombosis in childhood has come an appreciation of the impact of thromboembolic events upon children. In part, the greater awareness of childhood thrombosis reflects improved diagnostic imaging, which allows more sensitive identification of clots in infants and children. At the same time, advances in supportive care have meant that more children are being exposed to, surviving and living with thromboembolic risk factors and complications than previously. Because data regarding pediatric thrombosis have been lacking, treatment strategies have been extrapolated from experience in adults. This approach, however, results in diagnostic and therapeutic pitfalls. An understanding of issues unique to pediatric thrombosis is required; recent insights and the ongoing challenges are reviewed. PMID- 12529738 TI - Diacylglycerol kinases: regulation and signaling roles. AB - Diacylglycerol kinases (DGKs) are thought to attenuate diacylglycerol signals by converting diacylglycerol to phosphatidic acid. The nine mammalian diacylglycerol kinases that have been identified are widely expressed, but each isoform has a unique tissue and subcellular distribution. The activity of DGKs is regulated by mechanisms that can modify their access to diacylglycerol, affect their activity, or alter their ability to bind to other proteins. Although little is known of the specific function of DGKs in platelets, they likely influence actin reorganization and other signaling events requiring diacylglycerol. PMID- 12529739 TI - Intracranial hemorrhage in systemic lupus erythematosus associated with an autoantibody against actor XIII. AB - Intracranial hemorrhage in a young woman with systemic lupus erythematosus necessitated two surgical evacuations. In the absence of a family history of bleeding, clot solubility in urea suggested a factor XIII (FXIII) inhibitor. The patient's IgG bound well to the virgin and the thrombin-modified zymogen ensemble (A(2)B(2) and A(2)'B(2)) and to the free rA(2) but reacted poorly with the thrombin-modified rA(2)'. Since the IgG did not block the thrombin-catalyzed proteolysis of A subunits nor the dissociation of the A(2)'B(2), its action might be to interfere with the release of activation peptides from the thrombin-cleaved zymogen, hindering the conformational change necessary for generating FXIIIa. Treatment with cryoprecipitate and cyclophosphamide arrested the hemorrhage and almost neutralized the antibody so that the patient's clot became insoluble in urea and showed a close to normally crosslinked gamma-gamma and alpha(n) fibrin chain profile. Nevertheless, she still has detectable anti-FXIII antibody and may be at risk for hemorrhage. PMID- 12529740 TI - Pretreatment with oral anticoagulants decreases platelet activation in patients before and after percutaneous coronary intervention. AB - BACKGROUND: Platelet activation plays a major role in acute vessel closure after coronary angioplasty. In the randomized Balloon Angioplasty and Anticoagulation Study (BAAS), pretreatment with oral anticoagulants in addition to aspirin resulted in a 47% reduction of acute complications as compared with aspirin alone. This result may suggest a direct effect of oral anticoagulants on platelet activation. METHODS AND RESULTS: Patients were randomized to aspirin alone (group A, n = 26) or to aspirin plus oral anticoagulants started one week before angioplasty (group B, n = 26). Platelet response tests were performed 1 hour before (baseline) and 1 hour after intervention and on day 1. Platelet activation was measured by flow cytometry, as the number of antibody-positive platelets per 10,000 counted. Platelet function was evaluated with use of the PFA-100 analyzer. In group B, the median number of P-selectin-positive platelets was lower before (28 vs. 54, P = 0.018) and after (13 vs. 24, P = 0.377) angioplasty than in group A. Also the median decrease in the number of P-selectin-positive platelets during angioplasty was lower in group B (delta = 4) than in group A (delta = 30, P = 0.022). No further significant change was observed in platelet activation on day 1 in the two groups. The ability of platelets to become stimulated as measured with the PFA-100 analyzer was not affected by oral anticoagulants. CONCLUSIONS: Pretreatment with oral anticoagulants resulted in less activated platelets before and after coronary angioplasty, which is in agreement with its clinical effect of reducing procedural complications. Platelet function was not affected by oral anticoagulants. PMID- 12529741 TI - Thrombin-induced hyperactivity of platelets of young stroke patients: involvement of thrombin receptors in the subject-dependent variability in Ca2+ signal generation. AB - Activated platelets are implicated in the development of premature arterial vascular diseases, in particular ischemic stroke. Since elevated cytosolic [Ca(2+)](i) is an integrative marker of platelet activation, we determined the generation of Ca(2+) signal in stimulated platelets from 26 young patients recuperating from stroke, 20 patients with symptomatic peripheral arterial disease, and 56 healthy volunteers. Even in the presence of aspirin, the platelets from various individuals showed highly different thrombin-induced Ca(2+) responses. On average, the thrombin-induced Ca(2+) response was increased for platelets from either patient group in comparison to the controls (P <0.04). Relatively more stroke patients had high-responsive platelets (27%, 7/26) than patients with peripheral arterial disease (10%, 2/20) or healthy subjects (4%, 2/56). The average prothrombinase activities of platelets from patients and controls were similar, but 3 out of 6 patients with increased thrombin-induced Ca(2+) responses also exhibited high prothrombinase activity. In a follow-up study, the subject-dependent thrombin-induced Ca(2+) response was found to correlate strongly with the platelet response to protease-activated receptor 1 (PAR1) agonist (r = 0.91), but was not linked to the Pl(A1/2) polymorphism. It is concluded that a significant part of young patients with stroke have platelets with hyperactivity toward thrombin, which is not normalised by aspirin treatment. Furthermore, the subject-dependent variation in thrombin-induced signalling is likely to involve PAR1-mediated platelet activation. PMID- 12529742 TI - Should patients with deep vein thrombosis alone be treated as those with concomitant asymptomatic pulmonary embolism? A prospective study. AB - BACKGROUND: The established initial treatment of patients with deep vein thrombosis (DVT) or pulmonary embolism (PE) consists of the administration of subcutaneous, weight adjusted, low-molecular weight heparin (LMWH). However, the use of the same LMWH dosages for patients with either DVT or PE is not supported by data from comparative studies. PATIENTS AND METHODS: 1,000 consecutive patients with acute, proximal DVT were prospectively evaluated. All patients underwent a ventilation-perfusion lung scan on admission, and remained in hospital for at least 7 days. Patients with silent PE received once daily 10,000 to 15,000 IU subcutaneous LMWH dalteparin according to body weight for 7 days, and then vitamin K antagonists. Patients with DVT alone received LMWH in a fixed dose of 10,000 IU once daily for at least 5 days, and then vitamin K antagonists. The rate of both, major bleeding and symptomatic PE episodes during the 7-day study period was evaluated. RESULTS: Thirteen patients (1.3%) developed recurrent PE (1 died) and 16 patients (1.6%) had major bleeding (7 died). Recurrent PE was significantly more common in patients with silent PE (9 of 258 patients, 3.5%) than in those with DVT alone (4 of 742 patients, 0.5%. Odds ratio: 6.5; p <0.001). There were no significant differences in bleeding rate between patients with silent PE and those with DVT alone. However, the use of a fixed 10,000 IU dose in patients with DVT alone led to a significantly lower bleeding rate in patients weighing over 70 kg: 1 of 349 patients (0.3%) as compared to 9 of 393 patients (2.3%) in those weighing less than 70 kg (odds ratio: 0.12; p = 0.018). CONCLUSIONS: Fixed-dose 10,000 IU of LMWH dalteparin once daily proved to be both effective and safe in patients with DVT alone. The observed recurrence rate of 0.5% in these patients compares favourably with the 3.5% rate in patients with silent PE. Furthermore, this fixed-dosage was also safe. Patients weighing over 70 kg had a significant decrease in the rate of major bleeding, and no compensatory increase in the rate of recurrent PE. PMID- 12529743 TI - Combined activation of coagulation and inflammation has an important role in multiple organ dysfunction and poor outcome after severe trauma. AB - We tested the hypothesis that activated neutrophil-endothelial cell interaction in DIC can cause endothelial injury contributing to multiple organ dysfunction syndrome (MODS) and a poor outcome after trauma. Fifty-eight severe trauma patients, 29 with DIC and 29 without DIC were studied. Serial levels of soluble L , P-, and E-selectins, ICAM-1, VCAM-1, thrombomodulin, and neutrophil elastase were measured on days 0-4 after trauma. The numbers of systemic inflammatory response syndrome (SIRS) criteria that patients met were determined, simultaneously. In the DIC patients, higher DIC scores, lower platelet counts, and a longer duration of SIRS were found compared with the non-DIC patients. The incidence of ARDS and MODS were higher in patients with DIC than in those patients without DIC, and the DIC patients had poor outcome. Soluble L-selectin (sL-selectin) level on Day 1 in the DIC patients who died was markedly lower than those in the non-DIC patients. The levels of sP- and sE-selectins, sICAM-1, and sVCAM-1 were more elevated in the patients with DIC than in those without DIC on days 2 to 4. Neutrophil elastase and sThrombomodulin levels in the DIC patients persistently increased during the study period compared to those in the non-DIC patients. Maximum DIC scores in the DIC group showed good correlations with peak levels of sICAM-1, sVCAM-1, neutrophil elastase, sThrombomodulin, and the number of dysfunctioning organs. Highly activated and sustained inflammation caused by neutrophil-endothelium interaction in DIC gives rise to MODS and poor outcome in patients with severe trauma. These results suggest a close relationship between inflammation and thrombosis in posttrauma DIC. PMID- 12529744 TI - Clustering of thrombotic factors with insulin resistance in South Asian patients with ischaemic stroke. AB - We aimed to investigate significant correlations of insulin resistance with thrombotic factors in South Asians with stroke. Correlations of Homeostasis Model Assessment (HOMA)(as a surrogate of insulin resistance) were analysed with 6 thrombotic factors in 140 South Asian patients with a history of confirmed (by computerised tomography) ischaemic stroke. Age and sex adjusted HOMA was correlated to waist-hip ratio (r = 0.38, p = 0.0001), triglycerides (r = 0.22, p = 0.03), systolic blood pressure (r = 0.21, p = 0.04), tissue plasminogen activator (t-PA) (r = 0.22, p = 0.04); plasminogen activator inhibitor 1(PAI-1) (r = 0.26, p = 0.02); fibrinogen (r = 0.25, p = 0.02); and factor VII antigen (r = 0.21, p = 0.06). On regression analysis, with HOMA as dependent variable and significant correlates as independent variables in the model, HOMA was independently associated with PAI-1 antigen. There is extensive clustering of metabolic and thrombotic factors with insulin resistance in South Asian patients with ischaemic stroke, which may contribute to high prevalence of vascular disease in this population. PMID- 12529745 TI - Coagulation factor VII activity, Arg/Gln353 polymorphism and features of insulin resistance in first-degree-relatives of South Asian patients with stroke. AB - The aim of the study was to determine associations of factor VII:C levels in 140 South Asian stroke subjects and 143 first-degree relatives versus age-sex matched 146 control subjects without a personal or a family history of stroke subjects living in UK. There were no significant differences in Factor VII:C levels or FVII Msp I gene polymorphism (Arg-Gln 353) R353Q genotype frequency between the groups. R353Q genotype determined Factor VII:C levels in all the three groups. Factor VII:C levels correlated with triglycerides (patients, r = 0.23; relatives r = 27; control subjects, r = 0.24) and plasminogen activator inhibitor activity (patients, r = 0.30; relatives r = 0.22; control subjects r = 0.20) in all the three groups, but with insulin only in patients (p = 0.19). Circulating levels of Factor VII:C are determined by R353Q genotype and cluster with other risk factors associated with insulin resistance in South Asian ischaemic stroke patients, first-degree relatives and control subjects but are not related to stroke or a family history of stroke. PMID- 12529746 TI - Arterial intima-media thickness and its relationship with cardiovascular disease and atherosclerosis: a possible contribution of medium-sized arteries. AB - The intima-media thickness (IMT) of the arterial wall, measured by B-mode ultrasonography, has been related to cardiovascular disease and atherosclerosis in many studies, most of which have considered carotid and common femoral arteries. No study has related the IMT of medium-sized arteries (e.g. brachial) to the atherosclerotic process. The aim of this study was to evaluate whether more peripheral arteries (e.g. brachial, superficial femoral and popliteal) represent a good model in studies of arterial wall IMT, as well as carotid artery. Twenty-six male patients with demonstrated coronary artery disease (median age 63 yrs; range 41-70) and twenty-four male controls (median age 62 yrs; range 53-74) were studied. The far-wall IMT of the common carotid, brachial, superficial femoral and popliteal arteries was measured by high-resolution B-mode ultrasonography. Its relationship with ischemic cardiovascular disease and the presence of atherosclerotic plaques in all the arteries was also evaluated. Mean IMT values were higher in patients than controls, more so in superficial femoral artery (IMT difference 0.08 mm [95% confidence interval 0.02 - 0.14]) and popliteal artery (0.08 [0.01-0.15]) than in common carotid artery (0.02 [-0.07 0.11]) or brachial artery (0.01[-0.01-0.03]). The difference did not change after adjustment for age. The prevalence of plaques was consistently higher in patients than controls, except for brachial artery, in which no plaque was found either in patients and controls. In all the arteries except brachial we found an association between increase in IMT and number of plaques. Age was strongly related to the presence of plaques in the carotid artery, and less markedly in superficial femoral and popliteal arteries. The brachial artery does not seem a good model to study atherosclerosis by ultrasound measurements of arterial IMT, whereas superficial femoral and popliteal arteries might be chosen for these studies besides carotid artery. PMID- 12529747 TI - Targeted inactivation of the mouse locus encoding coagulation factor XIII-A: hemostatic abnormalities in mutant mice and characterization of the coagulation deficit. AB - Blood coagulation factor XIII (FXIII) promotes cross-linking of fibrin during blood coagulation; impaired clot stabilization in human genetic deficiency is associated with marked pathologies of major clinical impact, including bleeding symptoms and deficient wound healing. To investigate the role of FXIII we employed homologous recombination to generate a targeted deletion of the inferred exon 7 of the FXIII-A gene. FXIII transglutaminase activity in plasma was reduced to about 50% in mice heterozygous for the mutant allele, and was abolished in homozygous null mice. Plasma fibrin gamma-dimerization was also indetectable in the homozygous deficient animals, confirming the absence of activatable FXIII. Homozygous mutant mice were fertile, although reproduction was impaired. Bleeding episodes, hematothorax, hematoperitoneum and subcutaneous hemorrhage in mutant mice were associated with reduced survival. Arrest of tail-tip bleeding in FXIII A deficient mice was markedly and significantly delayed; replacement of mutant mice with human plasma FXIII (Fibrogammin P) restored bleeding time to within the normal range. Thrombelastography (TEG) experiments demonstrated impaired clot stabilization in FXIII-A mutant mice, replacement with human FXIII led to dose dependent TEG normalization. The mutant mice thus reiterate some key features of the human genetic disorder: they will be valuable in assessing the role of FXIII in other associated pathologies and the development of new therapies. PMID- 12529748 TI - Multicentre evaluation of IL Test Free PS: a fully automated assay to quantify free protein S. AB - Deficiency of the anticoagulant vitamin K-dependent protein S (PS) is associated with increased risk of venous thrombosis. In human plasma, PS circulates in two forms: as free protein (free PS) and PS bound to C4b-binding protein (C4BP), a regulator of the complement system. Assays for free PS have higher sensitivity and specificity for protein S deficiency than assays for total protein S. We have extensively evaluated the analytical performance of a novel assay for free PS, the IL Test Free Protein S, which takes advantage of the affinity of C4BP for free PS, and compared its performance to existing methods. IL Test Free Protein S is a rapid, fully automated turbidimetric assay consisting of two reagents: a C4BP coated latex and an anti-PS monoclonal antibody coated latex. The test range, precision and linearity were adequate and the assay tolerated high concentrations of interfering substances of clinical significance. The reference range agreed with previously published studies. The analysis of 903 patient samples belonging to 20 different clinical categories with the new assay yielded free PS results that agreed well with those obtained using the assays established in the participating laboratories. The study demonstrated the IL Test Free Protein S to be rapid, reliable and easy to perform. PMID- 12529749 TI - Rescue of prothrombin-deficiency by transgene expression in mice. AB - Prothrombin has diverse biological functions in addition to its well established role in blood coagulation. In order to study these functions in more detail mouse model systems are needed. Since deficiency of prothrombin in mice results in partial embryonic lethality and neonatal death, alternative approaches are required to study the biology of prothrombin in the adult mouse. The liver is the major site of synthesis of prothrombin and therefore liver-specific promoters were used to express prothrombin in transgenic mice. Mice generated from crosses with these transgenic mice and mice hemizygous for the knock-out allele were used to test whether liver-specific expression is sufficient to correct the phenotype of null mice and whether liver-specific expression is sufficient for the development and survival of mice to adulthood. The mouse albumin promoter/enhancer was used initially for transgene expression without success in obtaining transgene positive, endogenous prothrombin null mice. Two lines of transgene positive, endogenous prothrombin deficient mice were obtained using the mouse transthyretin (TTR) promoter/enhancer driving expression of a human prothrombin cDNA. One line was able to rescue both the embryonic and the neonatal lethality while the other line was only able to correct the embryonic lethality. Expression of prothrombin was restricted to the liver and stomach in one line and to the liver, pancreas, stomach and kidney in the other line of mice. Thrombin activity for one line was determined to be at 5-10% of wildtype levels. These mice developed normally and did not have spontaneous bleeding events unless traumatized. Therefore, transgenic expression of human prothrombin is sufficient for the rescue of the lethality found for prothrombin deficiency in mice. PMID- 12529750 TI - European Concerted Action on Anticoagulation (ECAA): International Normalized Ratio variability of CoaguChek and TAS point-of-care testing whole blood prothrombin time monitors. AB - The object was to assess the variability in displayed International Normalised Ratio (INR) between monitors of the same manufacture using whole blood samples from the same subjects. Two brands of monitor, CoaguChek Mini and the TAS PT-NC were tested. 14 instruments of each brand were tested on the same day at the same laboratory by the same operator using identical blood samples to avoid between centre differences in samples and operator technique. Whole blood samples from two normal donors and four coumarin-treated patients were tested to assess between-instrument variability of INR. Results have been coded. There was a much wider dispersion of INR on Brand B than on Brand A. One Brand A instrument failed to give a result with one of the two whole blood samples from one patient. One Brand B monitor gave an aberrant result with one of the samples from a normal subject. On both brands of monitor, INR variability appeared to be due mainly to duplication differences rather than between-instrument variability on both normal and coumarin whole blood samples. PMID- 12529751 TI - Localization and identification of thrombin and plasminogen activator activities in model human thrombi by in situ zymography. AB - Human thrombi vary in their susceptibility to lysis and this is clinically important. Several potential contributory factors were examined in this study by using model thrombi, created under flow; these provide a robust, reproducible and easily-manipulated system. Here we identify the plasminogen activators (PA) active in model thrombi of known age and define the cellular and plasma contribution to activity in different areas. The cell-rich head of model thrombi had strong thrombin and PA activity, with coagulant activity also at the tail. Thrombin activity decreased as model thrombi were aged. PA activity in the thrombus head also decreased on ageing of thrombi but activity emerged around the thrombi, including the tail. Activity in the head of fresh model thrombi was primarily due to uPA, with some contribution from tPA. Experiments with thrombi prepared from platelet-rich plasma and added leucocytes showed that uPA activity at the head of fresh thrombi was derived from PMN. Older thrombi had tPA activity around the tail of the thrombus; this activity occurred in the absence of cells. This study highlights the importance of PMN-derived uPA activity in the lysis of fresh thrombi, with activity originating in the leucocyte-rich head. It also shows that thrombi are dynamic structures in which fibrin can be repeatedly laid down and lysed, observations that are relevant to therapeutic lysis and potential rethrombosis. PMID- 12529753 TI - Combined effects of eptifibatide and anticoagulants: differences between LMWH and UH or rH in thrombin generation inhibition but not in platelet aggregation inhibition. AB - Aim of our study was to investigate effects of eptifibatide and anticoagulants on platelet aggregation and thrombin generation under low and high coagulant challenge in tissue factor-activated platelet rich plasma using a model allowing simultaneous determination of the time course of platelet aggregation and thrombin generation. Eptifibatide exerted a dose-dependent anti-aggregating effect under both high and significantly stronger under low coagulant challenge. Combination of eptifibatide and anticoagulants resulted in significant additive prolongation of the lag phase until the onset of platelet aggregation, more pronounced under low coagulant challenge. Under high, but not under low coagulant challenge combination of eptifibatide and anticoagulants had a significant synergistic inhibitory effect on platelet aggregation. Under low coagulant challenge combination of eptifibatide with LMWH, but not with UH, or rH, resulted in significantly reduced thrombin potential, F 1+2 generation, and FXa formation compared to measurements in the absence of eptifibatide. We demonstrate a synergistic effect of eptifibatide and anticoagulants on platelet aggregation inhibition and an additional inhibitory effect of LMWH and eptifibatide on thrombin generation. Our results support the notion that combination of eptifibatide and anticoagulants might be beneficial in atherosclerotic disease to palliate the thrombogenic potency of ruptured atherosclerotic plaques. PMID- 12529752 TI - A fully recombinant partial prothrombin complex effectively bypasses fVIII in vitro and in vivo. AB - The development of inhibitory antibodies is a serious complication in hemophilic patients, severely compromising therapeutic success. Bleeding episodes in affected patients are controlled by treatment with a plasma-derived prothrombin complex concentrate (PCC), activated PCC (APCC) or recombinant activated factor VII. We hypothesized that a recombinant two-component agent consisting of recombinant prothrombin (rfII) and activated factor X (rfXa) would have substantial fVIII bypassing activity and could be a safe alternative therapeutic option. To test this hypothesis we assembled an agent in vitro solely consisting of rfII and rfXa at a molar ratio of 37,500:1. These factors are believed to be responsible for the activity of APCC preparations. Recombinant fX, used as the source for fXa generation, and rfII were purified from serum-free and protein free conditioned media of stably transfected CHO and BHK tissue culture cells, respectively. Activation of rfX to rfXa was accomplished by the plant protease ficin, obviating the need for a protease derived from a human or animal source. We found that in vitro the complex reduced the abnormally prolonged activated partial thromboplastin time (APTT) of a high-titer fVIII inhibitor plasma similar to an APCC preparation. Furthermore, addition of increasing amounts of rfII/rfXa to inhibitor plasma resulted in a linear dose-dependent increase in the rate of thrombin generation. In a rabbit fVIII inhibitor model, treatment with rfII/rfXa statistically significantly reduced the intensity of the abnormal cuticle bleeding. In the Wessler test, rfII/rfXa showed no thrombogenicity. These data show that a well-defined, particularly safe and efficacious agent with fVIII bypassing activity can be generated from recombinant fII and fXa. PMID- 12529754 TI - Thrombin activatable fibrinolysis inhibitor (TAFI) levels in patients with coronary artery disease investigated by angiography. AB - Due to its role in the balance between coagulation and fibrinolysis, thrombin activatable fibrinolysis inhibitor (TAFI) may be involved in the development of cardiovascular diseases. We studied 362 patients with coronary artery disease (CAD) and 134 control subjects free of CAD, both groups investigated by angiography. TAFI antigen levels were determined in venous and intracoronary plasma samples and were related to metabolic and hemostatic risk factors and extent of coronary atherosclerosis. Venous TAFI levels tended to be higher in CAD patients compared to controls, whereas this difference was significant in intracoronary samples. A subgroup of patients who had not experienced acute myocardial infarction or undergone previous cardiac interventions showed significantly higher TAFI levels in both venous and intracoronary plasma samples. TAFI levels correlated with acute phase reactants indicating a role for TAFI in inflammation. However, TAFI levels did not correlate with extent of coronary atherosclerosis and among the classical cardiovascular risk factors TAFI levels only correlated with total cholesterol and fibrinogen concentration. Our results suggest that TAFI might be a risk factor for the development of CAD. PMID- 12529755 TI - A novel hemizygous Bernard-Soulier Syndrome (BSS) mutation in the amino terminal domain of glycoprotein (GP)Ibbeta--platelet characterization and transfection studies. AB - Glycoprotein (GP) Ib-V-IX is a unique adhesion receptor complex on platelets. Mutations in GPIbalpha, Ibbeta, and IX can lead to the rare bleeding disorder, Bernard-Soulier Syndrome (BSS). Here, we report a novel hemizygous variant of BSS in which Pro29 in one GPIbbeta allele is substituted by a Leu (GPIbbeta:P29L). Fluoresence in situ hybridisation revealed that the 22q11 locus was deleted from the homologous chromosome. The pedigree was determined and revealed inheritance of the GPIbbeta:P29L allele from the father. Flow cytometry with a range of antibodies detected no expression of GPIb-V-IX on the surface of the patient's platelets. Western blotting revealed an absence of GPIbalpha and GPIbbeta from platelet lysates. Co-expression of GPIbbeta:P29L with normal GPIbalpha and GPIX in a heterologous cell system confirmed that the mutant subunit did not support surface expression of the complex. Interestingly, residual expression of GPIbbeta:P29L anchored in the plasma membrane alone was now seen. This novel BSS mutation expressed in heterologous cells is in agreement with recent in vitro evidence that the correct conformation of the amino terminal region of GPIbbeta is required for normal expression of the intact receptor complex. PMID- 12529756 TI - Transfer of tissue factor from platelets to monocytes: role of platelet-derived microvesicles and CD62P. AB - Tissue factor (TF) is the most important initiator of intravascular coagulation. Platelets contribute to TF exposure on monocytes, but the mechanism is not completely understood. Here we examined the possibility that platelets may release TF that can be transferred to monocytes by platelet-derived microvesicles. When human citrated platelet-rich plasma was incubated with collagen there was an increase in the plasma levels of TF and CD62P. Incubation of plasma obtained from collagen-stimulated PRP with a sediment of red and white blood cells resulted in an increase in the number of monocytes that express TF, CD62P and the platelet-specific antigen CD42a on their surface. This transfer of platelet-derived antigens to monocytes was reduced when CD62P was blocked by a specific antibody or when platelet-derived microvesicles were removed from the plasma either by high speed centrifugation (17,500 x g for 30 min) or by filtration (pore size 0.2 microm). The data indicate that platelet-derived microvesicles that are released from collagen-stimulated platelets may carry TF, CD62P and CD42a and may transfer these antigens to the surface of monocytes. The interaction of platelet-derived microvesicles with monocytes and the transfer of TF to monocytes strongly depend on CD62P. PMID- 12529757 TI - Pathogenic effects of anti-glycoprotein Ib antibodies on megakaryocytes and platelets. AB - Antibodies directed against the glycoprotein (GP) Ib have been identified as the potential cause of various platelet disorders: Immune thrombocytopenic purpura (ITP) may be caused by such autoantibodies; Anti-thrombotic drugs targeting GPIb also induce thrombocytopenia. In order to elucidate the potential mechanism(s) of the anti-GPIb effects, we have examined by electron microscopy (EM) the effect of several antibodies directed against GPIb and GPIIb-IIIa on human culture megakaryocytes (MK). Virtually all antibodies to GPIb enhanced the interaction of newly formed platelets with MK when compared to other antibodies. These effects were retrieved when antibodies were tested on platelets. We conclude that antibodies to GPIb can potentially inhibit platelet release by MK, and can also induce homotypic platelet adhesion. These results may have implications in the pathophysiology of thrombocytopenia and platelet recovery in ITP, and shed light on the pathological effect of anti-GPIb antibodies used as antithrombotic drugs. PMID- 12529758 TI - Flow conditions modulate homocysteine induced changes in the expression of endothelial cell genes associated with cell-cell interaction and cytoskeletal rearrangement. AB - In the present study we analyzed changes in the pattern of gene expression in endothelial cells (HUVEC) exposed to elevated homocysteine levels under flow conditions. RNA samples of untreated and homocysteine treated EC were analyzed using a human cDNA array containing 588 cDNAs relevant to the cardiovascular system. Exposure to homocysteine under flow resulted in altered expression of 8 genes, the alterations of 3 of which were further confirmed by RT-PCR analysis: upregulation of MCP-1 and of profilin-I, and downregulation of alpha-catenin. The increased expression of profilin-I and the decreased expression of alpha-catenin were also confirmed at the protein level by Western blot analysis. Furthermore, immunostaining for alpha-catenin demonstrated complete loss of the protein from intercellular junctions following exposure to homocysteine under flow. All these changes were not observed in cells exposed to homocysteine under static conditions. We show that under flow conditions, homocysteine modulates the expression of genes/proteins involved in actin cytoskeleton reorganization, as well as in the formation and/or maintenance of cell-cell junction, thus contributing to the morphological changes (e.g. cell retraction) reflecting endothelial cell injury. PMID- 12529759 TI - Soluble tissue actor interferes with angiostatin-mediated inhibition of endothelial cell proliferation by lysine-specific interaction with plasminogen kringle domains. AB - Experimental and clinical data suggest that tissue factor (TF), the major initiator of blood coagulation cascade, as well as proteases and components of the fibrinolytic system are involved in tumor growth at least in some solid tumors via effects on angiogenesis. Whereas the pro- and anti-angiogenic effects of the plasminogen/plasmin system and plasminogen kringle domains, respectively, are well characterized, the pathways responsible for the pro-angiogenic properties of TF remain poorly understood. To learn more about the biological significance of the recently described binding of plasminogen to the extracellular domain of TF, we examined the effects of soluble TF (sTF) on angiostatin-inhibited proliferation of endothelial cells. In solid phase binding assays, we found that sTF binds specifically to plasminogen, to the plasminogen kringle domains K1-3, K1-5, K4, as well as to mini-plasminogen. Inhibition of binding of plasminogen and its kringle domains to sTF by the lysine analog 6 aminohexanoic acid (AHA) suggests that lysine-binding sites are involved in plasminogen interaction with TF. Moreover, in the presence of sTF, the inhibitory effect of K1-5 on bFGF-mediated HUVEC proliferation was dose-dependently and saturably abolished. This suggests that TF can interfere with the antagonistic effect of K1-5 on endothelial cell proliferation. In contrast, sTF by itself had no effect on the endothelial cell proliferation. Whereas the interference of TF with K1-5-mediated effect was prevented by AHA, this lysine analog did not abolish the proliferation inhibition of K1-5. In conclusion, the binding of sTF to the plasminogen fragment K1-5 seems to antagonize the anti-angiogenic effects of this plasminogen fragment. PMID- 12529760 TI - Regulation by nitric oxide of endotoxin-induced tissue factor and plasminogen activator inhibitor-1 in endothelial cells. AB - The increase in nitric oxide (NO) production in lipopolysaccharide (LPS)-induced sepsis is thought to contribute to the development of shock. However, NO could also play an antithrombotic role. Little is known about the modulating effect of NO on the endothelial overexpression and production of tissue factor (TF) and plasminogen activator inhibitor-1 (PAI-1) occurring in endotoxemia. We analyzed the effect of N(G)-nitro-L-arginine-methyl-ester (L-NAME), an inhibitor of NO synthases, and S-nitroso-N-acetyl-D,L-penicillamine (SNAP), a NO donor, on the expression and synthesis of TF and PAI-1 by LPS-challenged human umbilical vein endothelial cells (HUVEC): L-NAME enhanced the increase in TF mRNA and antigen levels (P <0.05) observed in LPS-treated HUVEC; SNAP down-regulated the LPS induced TF increment (p <0.05). However, no effects of NO on regulation of the LPS-dependent increase in PAI-1 could be seen. Thus, NO could play an antithrombotic role in sepsis by down-regulating the endothelial overexpression and production of TF. PMID- 12529761 TI - Novel fibrinogen Bbeta gene mutation causing hypofibrinogenaemia. PMID- 12529762 TI - A study on the content of CSF vWF in patients with cerebral infarction at different location. PMID- 12529763 TI - Endothelial cell protein C receptor (EPCR) gene exon III, 23 bp insertion mutation in the Turkish pediatric thrombotic patients. PMID- 12529764 TI - Association between sensitivity for activated protein C (APC) and lipid or lipoprotein levels. PMID- 12529765 TI - Allelic discrimination of factor V Leiden using the GeneAmp 5700 Sequence Detection System. PMID- 12529766 TI - Absence of factor V Leiden mutation and low prothrombin G 20210 A mutation prevalence in a healthy Moroccan population. PMID- 12529767 TI - The impact of seasonal and meteorological conditions on the incidence of thrombotic haemodialysis shunt occlusions. PMID- 12529768 TI - Rebuttal: procoagulant activity of T lymphoblastoid cells. PMID- 12529770 TI - Comment: raised soluble P selectin and the risk of thrombosis in HIV-1 infection. PMID- 12529771 TI - [Results of C1-C2 transarticular screw fixation in a series of 20 patients]. AB - OBJECTIVE: C1-C2 transarticular fixation is an increasingly used surgical method of treating atlantoaxial instability. When properly performed, it can safely provide fusion rates near 100%, but techniques of fixation in this region allow only a small margin of error. We here report the results of C1-C2 transarticular fixation in a series of 20 patients in which different procedures were selected according to the presenting disorder in each case. METHODS: The study group included 9 men and 11 women with a mean age of 48 years (range 17 to 68 years). The causes of instabilities were rheumatoid arthritis in nine patients, type II and III Hangman's fracture of traumatic origin in nine (in association with other lesions in three cases), pseudoarthrosis after type II odontoid fracture in one, and type III complex C1 fracture in a patient with ankylosing spondylitis. Preoperative assessment included flexion and extension plain radiographs and computed tomography (CT) and magnetic resonance imaging (MRI) scans. Operations included transarticular screw fixation in all cases; in patients with rheumatoid arthritis it was associated with sublaminar fixation and bone grafting following Sonntag's technique in all but two cases. Postoperative results were evaluated in relation to the biomechanical stability and fusion was studied by flexion and extension radiographs and CT scans. Pain relief in patients with rheumatoid arthritis patients was assessed using a 0-10 visual analogic scale (V.A.S.). RESULTS: In the traumatic group, a consolidation of the fracture and radiologic stability was achieved in all cases. In patients with rheumatoid arthritis, pain improved in all but not the neurological deficit, and in all cases a C1-C2 biomechanical stability was reached despite interlaminar graft resorption in two (25%) cases. With respect to complications, a lesion of the vertebral artery occurred in one case, deviation of the screw in two cases, and breakage of the screw without clinical repercussion in two other cases. CONCLUSION: C1-C2 transarticular screw fixation was a useful technique to achieve satisfacory biomechanical stability in patients with atlatoaxial instability of both inflammatory and traumatic origin with a low rate of complications. PMID- 12529772 TI - Surgical treatment of trigonocephalies and associated hypoteleorbitism. AB - Premature closure of metopic suture is a relatively uncommon form of craniosyostosis with an estimated incidence of 0,3 per 1000 live births, comprising about 7% of surgical craniosynostosis referred to craniofacial centers. A broad phenotypical spectrum spreads from minor metopic ridges to severe trigonocephaly with pterional indentation supraorbital bar retrusion, temporal and parietal compensating bossings and hypotelorism. Most of the cases arise spontaneously although autosomal dominant inheritance has been described and association with cromosomal abnormalities and different syndromes has been widely reported. Surgical correction has been attempted with good cosmetic results using several variations of the standard frontoorbitary advance. However there is still a number of questions to be solved in relation to this entity, mainly on its pathogenesis, but also on its development, natural history and treatment. Direct surgical approach to associated hypotelorism is a matter of argument when considering the reestablishment of normal interorbitary distances. We have conducted a retrospective analysis of our serie consisting of twenty eight cases of trigonocephalies. Surgical correction of hypotelorism was attempted in eleven cases while the resting seven children remained "not treated". The objective was to review the functional outcome and cosmetic results comparing the different techniques applied to the frontal bone and to observe evolution of the hypoteleorbitism after the treatment with or without osteotomies and grafting of the nasoethmoidal area. PMID- 12529773 TI - [Surgical management of anterior communicating artery aneurysms based on computed tomographic angiography with three-dimensional reconstruction and without preoperative angiography]. AB - OBJECTIVE: To demonstrate the usefulness of threedimensional computed tomographic angiography (CT- 3D-angiography) in the microsurgical management of aneurysms of the anterior communicating artery (AComA). MATERIALS AND METHODS: A total of 28 consecutive patients with ruptured aneurysms of the AComA diagnosed by means of CT-3D-angiography and without preoperative angiography were operated on. The findings of the CT-3D-angiography, microsurgical exploration and clinical data were evaluated. RESULTS: There were no false positive findings nor false negative findings in the diagnosis of the AComA aneurysms. The global sensibility of the examination was 87.9%. The CT-3D-angiography study shows a left A1 segment dominance in 53.6% of cases, a right A1 dominance in 14.3% of cases and both A1 segments of the same diameter in 32.1%. Aneurysms growing on the traject of the AComA were associated with both A1 segments of the similar diameter and an AComA traject pararell to the transverse plane. Aneurysms implanted on the A1-A2 junction were associated with a dominant homolateral A1 segment and an oblique AComA traject. Microsurgical management of the lesions was done a mean of 3.7 days after bleeding. CONCLUSION: The study of patients with acute subarachnoid hemorrhage with CT-3D-angiography allows a reliable diagnosis of AComA aneurysms. The examination gives some anatomical data that allow the study of the hemodinamic changes involved in the development of the aneurysms. Moreover, provides usefull information for the microsurgical clipping. CT-3D-angiography allows to improve some health indicators but its impact in the final result of the patients needs more clinical data. PMID- 12529774 TI - [Anterior sacral meningocele with Currarino's syndrome: report of two cases]. AB - The anterior sacral meningocele is the congenital hernia of the thecal sac into the pelvis through a bone defect of the anterior wall of the sacrum. The association of anorectal anomaly, anterior sacral defect and presacral mass, known as Currarino's triad, can be diagnosed during the first decade of life in the eighty per cent of the cases However, in the incomplete syndrome, the symptoms appear in adults. In order to avoid late complications and due to the clear genetic origin of this uncommon pathology, the investigation of the family is of paramount importance for the early diagnosis and treatment. We review the literature and add two cases of anterior sacral meningocele with rectal fistula and incomplete Currarino's syndrome. In one of the cases the pathology is also present in another member of the same family and is complicated with meningitis. The importance of early diagnosis and treatment of these malformations and the need for fammily genetic investigation are discussed. PMID- 12529775 TI - [Spinal extradural lipomatosis. Revision of 108 cases. Case induced by exogenous contribution of ACTH]. AB - Authors present a male patient with Spinal Extradural Lipomatosis, previously treated of a cerebral astrocytoma with surgery and radiotherapy, after which he received ACTH for a long period of time. Clinical manifestations were rachialgia, paraparesia with pain and dysestesias in both lower extremities. Diagnosis was carried out by Magnetic Resonance imaging. After a progressive withdrawal of the treatment with ACTH, the patient achieved a complete recovery and neuroimaging studies showed the dissappearance of the compression caused by the lipomatosis. We carry out a revision of the literature showing data we consider of interest derived from the wide series subjected to study. PMID- 12529776 TI - Redundant ventriculomegaly associated with brain abscess. AB - OBJECTIVE: The aim of this clinical retrospective review was to emphasize the recently observed high incidence of ventriculomegaly, a wellknown complication of brain abscesses. MATERIALS AND METHODS: Among twelve brain abscess cases operated on in our institution during five years, the most common surgical procedure was burr-hole aspiration applied in ten of the cases, while in the remaining two a standard craniotomy and total excision was performed. Ventriculomegaly was assessed by the "Evans' Ratio" values calculated using the frontal ventricular horn measurements made on axial tomographic images. RESULTS: The signs, symptoms, radiological findings and clinical features of the patients were in accordance to the pertinent literature. The only finding worth commenting on was the surprisingly high incidence of ventriculomegaly (5/12) and periventricular edema (4/12). Concerning the outcome, there was only one death. CONCLUSIONS: The "Evans' Ratio" either worsened or remained the same in four out of five cases during the early postoperative period (ranging from one to three months) and it got better in one case in the late postoperative control (1.5 years). Furthermore, periventricular edema persisted just in the same manner in three of the four cases. PMID- 12529777 TI - Isolated primary Langerhans' cell histiocytosis of central nervous system. AB - Solitary eosinophilic granuloma that involves the CNS is an uncommon lesion and most frequently affects the hypothalamus. We report a new and rare case of solitary eosinophilic granuloma of the left temporal lobe in a patient without systemic disease. The diagnosis was confirmed by electron microscopy and immuohistochemical techniques. The treatment of choice is surgical resection. There is a better prognosis in comparison with cases accompanied by systemic disease. PMID- 12529778 TI - Intratumoural bleomycin as a treatment for recurrent cystic craniopharyngioma. Case report and review of the literature. AB - INTRODUCTION: The majority of craniopharyngioma (CF) have a cystic component and only 10% are completely solid. In tumors with a large cystic component, stereotactic drainage or instillation of radioctive and/or chemotherapeutic agents have been used. Only several authors have reported the use of bleomycin for the treatment of cystic CF. CASE REPORT: The authors present the case of a nineteen years old patient with a recurrent cystic CF who was treated with intratumoral injections of bleomycin. The patient had been operated on three times before because of regrowth of the tumor. This last time he had a severe disturbance of his visual acuity and a huge regrowth of the cystic CF. An intracystic catheter stereotactically placed was connected to an Ommaya reservoir and, after assuring the impermeability of the cyst, bleomycin was administered through the reservoir up to a total dose of 45mg distributed in six doses. No complications were detected during and after the procedure. A MR performed 4 months after treatment showed a clear reduction in the size of the cyst but 10 months later a new regrowth of the cyst was detected by MR with no new signs or symptoms. A total dose of 30 mg divided in six doses was administered. No complications occurred. The MR 18 months after the first treatment showed the reduction in size of the tumor. The ophtalmological study showed almost normal visual acuity in both eyes. DISCUSSION: Although there is not an stablished protocol for the indication and the form of application of intracystic bleomycin, results with this treatment for cystic CF seem good in the literature. However, the risk of local complications after the administration of intratumoural bleomycin in these patients is around 10%, and some fatal toxic reactions have been recently reported. CONCLUSION: Intracystic administration of bleomycin is a valid option as adjuvant therapy for CF in patients with recurrences that are not surgical candidates because of the high risk of complications. The role of bleomycin as a primary treatment for CF and treatment protocols remain to be stablished with additional studies. PMID- 12529779 TI - [Spontaneous resolution of an acute subdural hematoma]. AB - Acute postraumatic subdural hematoma is known to have bad prognosis. Traditionally, its treatment consisted in urgent surgical evacuation. Rapid spontaneous resolution of the hematoma has been infrequently observed. We report a case of a patient with an acute traumatic subdural hematoma which disappeared without surgery. CT-scan showed a left-sided subdural hematoma with marked mass effect. A low density layer, which implied involvement of CSF in the subdural space, was observed between the hematoma and the inner wall of the skull. We think that the spontaneous resolution of the hematoma was due to the dilution and washing-out by the CSF through an arachnoid tear. Another suggested mechanism in the literature is blood redistribution in the subdural space. Selected patients with an acute subdural hematoma can be managed with conservative treatment. PMID- 12529780 TI - [Rapid spontaneous resolution of acute subdural hematoma]. AB - Acute subdural hematomas are usually neurosurgical emergencies, although a conservative therapy is indicated in selected cases. In some of these patients a progressive resolution is appreciated. However, rapid spontaneous resolution of an acute suddural hematoma is seldom reported. A patient with antecedent of chronic alcoholism and spontaneous resolution of acute subdural hematoma in less than 18 hours after the head injury is described. The possible mechanisms of this rapid resolution are discussed. A tear in the arachnoid with spilling of CSF into the subdural space and the effect of intracranial antihypertensive measures together with cerebral atrophy, are factors that possibly contribute to dilution and redistribution of blood with hematoma disappearing in CT scan. PMID- 12529781 TI - [Cemento-ossifying fibroma of the skull]. AB - Cemento-ossifying fibroma (cementoma) is benign fibro-osseous lesion of bone that generally is first seen during childhood or young adulthood as asymptomatic or painful intraosseous mass that commonly involved the maxilla. This paper describes the case of a young boy with a cemento-ossifying fibroma of the skull. PMID- 12529782 TI - [The evaluation of the neurosurgeon]. PMID- 12529783 TI - [Our problems]. PMID- 12529784 TI - [Informed consent]. PMID- 12529785 TI - Clinical features and molecular genetics of hereditary peripheral neuropathies. AB - Hereditary peripheral neuropathies are the most common monogenetically inherited diseases of the nervous system. The prevalence of the Hereditary Motor and Sensory Neuropathy Type 1A (HMSN 1A or Charcot-Marie-Tooth Neuropathy 1A, CMT1A) alone is estimated to be as high as 1/5000. In 1991, a duplication on chromosome 17p11.2 was identified as the causative genetic defect of CMT1A. Since then causative mutations in 17 genes have been identified. This review summarises the clinical and molecular genetic features of primary inherited neuropathies. It is aimed primarily at clinicians and geneticists. Therefore less emphasis is placed on the pathology and the (often unknown) underlying biological disease mechanisms. PMID- 12529786 TI - Cerebellar volumes in newly diagnosed and chronic epilepsy. AB - Cerebellar atrophy is assumed to be a common finding in patients suffering from epilepsy. Anticonvulsants as well as seizure activity itself have been considered to be responsible for it but many studies have addressed these questions in specialised centres for epilepsy thus having a referral bias towards patients with severe epileptic syndromes. The purpose of this study was: 1. To develop a quantitative method on 3D-MRI data to achieve volume or planimetric measurements (of cerebrum, cerebellum and cerebellar substructures). 2. To investigate the prevalence of cerebellar atrophy (and substructure atrophy) in a prospectively investigated population-based cohort of patients with newly diagnosed and chronic epilepsy. 3. To quantify cerebellar atrophy in clinic-based patients, who had had atrophy previously diagnosed on routine visual MRI assessment. 4. To correlate the measures of atrophy with clinical features in both patient groups. A total of 57 patients with either newly diagnosed or chronic active epilepsy and 36 control subjects were investigated with a newly developed semiautomated method for cerebral as well as cerebellar volume measurements and substructure planimetry, corrected for intracranial volume. We did not find any significant atrophy in the population-based cohort of patients with newly diagnosed epilepsy or with chronic epilepsy. Visually diagnosed cerebellar atrophy was mostly confirmed and quantified by volumetric analysis. The clinical data suggested a correlation between cerebellar atrophy and the duration of the seizure disorder and also the total number of lifetime seizures experienced and the frequency of generalised tonic-clonic seizures per year. Volumetry on 3D-MRI yields reliable quantitative data which shows that cerebellar atrophy might be common in severe and/or longstanding epilepsy but not necessarily in unselected patient groups. The results do not support the proposition that cerebellar atrophy is a predisposing factor for epilepsy but rather are consistent with the view that cerebellar atrophy is the aftermath of epileptic seizures or anticonvulsant medication. PMID- 12529787 TI - Clinical spectrum of pontine infarction. Clinical-MRI correlations. AB - We sought clinical and radiological findings of 150 consecutive patients with acute isolated pontine infarct who were admitted to our Stroke Unit over 6 years. In all patients CT, MRI and magnetic resonance angiography (MRA) were performed during the hospitalization. On clinico-radiological analysis regarding the pontine lesion boundaries there were five main clinical patterns that depended on the constant territories of intrinsic pontine arteries: (1). anteromedial pontine syndrome (58%) presented with motor deficit with dysarthria, ataxia, and mild tegmental signs in one third of patients; (2). anterolateral pontine syndrome (17%) developed with motor and sensory deficits in half of the patients, and were associated with tegmental signs (56%) more frequently than the anteromedial infarct syndrome; (3). tegmental pontine syndrome (10%) presented with mild motor deficits and associated with sensory syndromes, eye movement disorders and vestibular system symptoms including vertigo, dizziness and ataxia; (4). bilateral pontine syndrome (11%) consisted with transient consciousness loss, tetraparesis and acute pseudobulbar palsy; (5). unilateral multiple pontine infarcts (4%) were rarely observed, and were always associated with severe sensory-motor deficits and tegmental signs. In our series, there was no infarct in the extreme dorsal and lateral tegmental pontine territories which have been mostly associated with cerebellar infarctions. The main etiology of stroke was basilar artery branch disease (BABD) in 59 patients (39%), followed by small artery disease (SAD) in 31 (21%), large-artery disease of vertebrobasilar arteries in 27 patients (18%), cardioembolism in 12 (8%) and in 16 patients (11%) no cause of stroke was found. Our findings suggest that it is possible to identify clinical subgroups of pontine infarction, in which BABD and SAD were the most common causes of stroke. After an acute onset, outcome is in general excellent except in those with bilateral pontine lesions. PMID- 12529788 TI - Four year follow-up study after unilateral pallidotomy in advanced Parkinson's disease. AB - BACKGROUND: Pallidotomy is widely accepted as a treatment for patients with Parkinson's disease (PD) who have disabling symptoms that are inadequately controlled with pharmacotherapy. There are, however, only a few studies, with a small number of patients, showing evidence after prolonged periods of time about the clinical outcome after pallidotomy. METHODS: We have conducted a four-year follow-up study of 23 patients with PD who had undergone unilateral pallidotomy. Six patients were lost to follow-up. The remaining 17 were evaluated before surgery and at three months, one year, and four years after surgery, following standard protocols. RESULTS: At four years, patients still showed significant amelioration in contralateral and axial dyskinesias and off-period contralateral parkinsonian signs. Total motor UPDRS and Activities of Daily Living scores also remained improved four years after surgery. Ipsilateral and axial parkinsonian symptoms and Hoehn & Yahr and Schwab & England scales were not significantly changed from baseline values. The initial beneficial effects on motor fluctuations were not maintained at last visit. All patients considered themselves to be improved with respect to their clinical condition before surgery in a clinical global impression estimation done at the four-year follow-up. CONCLUSION: Unilateral pallidotomy can provide sustained improvement of contralateral parkinsonism and dyskinesias during at least four years. Early benefits observed in axial symptoms and motor fluctuations wane with time. On period akinesia worsened significantly in the limbs contralateral to pallidotomy after four years. PMID- 12529789 TI - Levodopa and bromocriptine in hypoxic brain injury. AB - BACKGROUND: Postanoxic encephalopathy is frequent in patients presenting with abrupt cardiac arrest or respiratory failure. Little is known about the effectiveness of oral medications on the cognitive and motor consequences. OBJECTIVE: To present data suggesting partial improvement after administration of levodopa/benserazide. METHODS: After observing partial benefit in one case, each patient admitted to rehabilitation following brain anoxia was systematically treated with levodopa/benserazide (200/50 to 400/100 mg/day), then bromocriptine (15 mg/day). RESULTS: In the first patient, brain anoxia was severe, with secondary agitation, quadriparesis, involuntary movements, inattention and communication disorders. Introduction of levodopa/benserazide resulted in reduction of agitation and involuntary movements and improvement of communication, thus facilitating care and rehabilitation efforts. A weaning test resulted in rapid worsening. The four following patients also presented with anoxia of variable severity. Marked improvement was observed in case 2, presenting with agitation, loss of orientation, amnesia, postural disorders, involuntary movements and dysphagia, with a withdrawal test resulting in immediate re-enhancement of symptoms. Modest improvement was observed in patient 3, who had hypokinesia, rigidity, adynamia, impaired attention, and reduced verbal fluency. Patient 4 presented with memory disorders without motor difficulties: mild improvement was observed in daily life and memory tests. In patient 5 who also presented with severe memory disorders, the benefit was absent. In each case, bromocriptine was introduced 3-4 weeks following levodopa, but without additive effect. Both treatments could be interrupted after a few months, without worsening. CONCLUSIONS: Levodopa and benserazide can be of benefit in the few months following brain anoxia, especially on some of the motor disorders and apathy, but the benefit is inconstant and modest on memory disorders. Anoxia could alter dopaminergic mesencephalic systems, which activate the striatal and mediobasal frontal cortex, and these disorders could be partially reversible by medical treatment. PMID- 12529790 TI - Three-dimensional analysis of nystagmus in benign paroxysmal positional vertigo. New insights into its pathophysiology. AB - Nystagmus is a rhythmical rotation of the eyeball. Its characteristics can be defined mathematically by the axis of rotation and the angular velocity around this axis. We analysed the axis of rotation for the nystagmus in benign paroxysmal positional vertigo (BPPV) to elucidate its pathophysiology. Thirteen patients with typical unilateral BPPV participated in the present study. The axis of rotation for the nystagmus was calculated from images recorded on digital videotape with an infrared CCD camera, using an algorithm that we developed. The patients' responsiveness to Semont's liberatory maneuver was also assessed.The results showed that patients could be assigned to one of two groups based on the rotation axis of the nystagmus. In one group of patients (n = 7; Group A), the axis of rotation was almost vertical to the plane containing the posterior semicircular canal (PSC) on the presumed affected side. In the other group of patients (n = 6; Group B), it was clustered around the naso-occipital axis. These results suggest that in the patients in Group A the responsible lesion is confined to the PSC, whereas the pathogenesis underlying the conditions in those in Group B is considered to be more complicated. This speculation was reinforced by the difference in responsiveness to Semont's liveratory maneuver. All of the patients in Group A showed remission of vertigo within 10 days (mean, 2.4 days) after onset of treatment using Semont's liberatory maneuver, whereas the patients in Group B required much longer periods of time for remission to occur (mean, 25.7 days). Our findings lead to the conclusion that by considering the spatial orientation of individual semicircular canals, in the patients in Group B the pathophysiology is not confined to the PSC, but most probably all three semicircular canals are involved. PMID- 12529791 TI - Effects of subthalamic nucleus stimulation on actual and imagined movement in Parkinson's disease : a PET study. AB - BACKGROUND: PET studies in moderately affected Parkinson's disease (PD) patients reveal abnormal cerebral activation during motor execution and imagery, but the effects of subthalamic nucleus (STN) stimulation are not well established. OBJECTIVES: to assess the effect of STN stimulation on cerebral activation during actual and imagined movement in patients with advanced PD. METHODS: seven severely affected PD patients treated with bilateral STN stimulation were studied with PET and H(2)(15)O. The following conditions were investigated: (1). rest; (2). motor execution of a sequential predefined joystick movement with the right hand and (3). motor imagery of the same task. Patients were studied with and without left STN stimulation while right stimulator remained off. RESULTS: Without STN stimulation, the primary motor cortex was activated only during motor execution whereas the dorsolateral prefrontal cortex (DLPFC) was activated only during motor imagery. An activation of the supplementary motor area (SMA) was seen during both motor execution and motor imagery. Left STN stimulation during motor execution increased the regional cerebral blood flow (rCBF) bilaterally in the prefrontal cortex including DLPFC, in the left thalamus and putamen. In addition, a reduction of rCBF was noted in the right primary motor cortex, inferior parietal lobe and SMA. Under left STN stimulation, during motor imagery, rCBF increased bilaterally in the DLPFC and in the left thalamus and putamen and decreased in the left SMA and primary motor cortex. CONCLUSION: STN stimulation during both motor execution and imagery tends to improve the functioning of the frontal-striatal-thalamic pathway and to reduce the recruitment of compensatory motor circuits notably in motor, premotor and parietal cortical areas. PMID- 12529792 TI - Tactile hallucinations in Parkinson's disease. AB - Hallucinations occur in up to 40% of patients with Parkinson's disease (PD) and are mainly of a visual nature. We prospectively studied 8 patients with PD and tactile hallucinations (TH). TH occurred with a clear sensorium, and were long lasting. In most cases they involved animals, were combined with other types of hallucinations occurring simultaneously (mainly formed visual hallucinations), and predominated in the evening and/or at night. Pharmacological and disease related factors, including a disorder in rapid-eye-movement sleep mechanisms, could play a part in the pathophysiology of these hallucinations. PMID- 12529793 TI - Spontaneous supratentorial intracerebral hemorrhage. Criteria for short-term functional outcome prediction. AB - BACKGROUND AND PURPOSE: To identify the significant prognostic factors, upon admission, and construct a set of criteria to predict short-term functional outcome of patients with intracerebral hemorrhage (ICH). METHODS: The records as well as the radiological findings, of 184 consecutive cases of spontaneous supratentorial intracerebral hemorrhage, that were treated medically, were reviewed. The hemorrhage was graded according to size, mass effect and intraventricular extension. Outcome upon discharge was scored using the modified Rankin Scale. A score of four or more was considered a poor outcome. Multivariate analysis was used to identify the factors associated with a poor outcome. RESULTS: Six significant and independent prognostic variables were identified: decreased level of consciousness, severe hemiparesis, age older than sixty, large hematoma size, midline shift and intraventricular extension on CT. These variables were scored systematically to produce the ICH criteria. The sum of these criteria yields a figure between zero and six termed ICH score. Patient grouping according to ICH score identified four distinctive, prognostic groups: I - score of zero to one, II - score of two, III - score of three and IV - score of four to six in which 82%, 53.7%, 23.3% and 0% achieved a good outcome respectively (P < 0.05 for all groups). CONCLUSIONS: The present study identified six independent admission criteria predicting the short-term functional outcome of ICH patients. Their sum may serve to predict the short-term functional outcome upon admission of medically-treated patients with spontaneous supratentorial intracerebral hemorrhage. PMID- 12529794 TI - Anti-Ri antibody positive opsoclonus-myoclonus in a male patient with breast carcinoma. AB - A 65-year-old male patient developed truncal ataxia, opsoclonus and myoclonus. In the serum anti-Ri antibodies were found, which led to the detection of a small adenocarcinoma of the breast. Other prominent clinical features were an excessive startle response and behavioral disorders, such as anxiety and impatience. These features suggest an immune response against both Nova-1 and Nova-2 antigens throughout the central nervous system. PMID- 12529795 TI - Ocular manifestation of primary nervous system lymphoma: what can be expected from imaging? AB - Primary ocular lymphoma, which affects the posterior parts of the eye, is an ocular manifestation of primary central nervous system lymphoma (PCNSL). It used to be the ocular disease with the shortest time of survival, even worse than ocular melanoma. Death ensues by CNS dissemination. Unfortunately, ocular lymphoma may be the initial manifestation of PCNSL and diagnosis is frequently difficult, even if vitreal biopsy is performed. Therefore, it should be determined whether cross sectional imaging may be helpful in detection and differential diagnosis of ocular lymphoma. MRI of seven patients (female = 6, male = 1, median age 62 years) with biopsy proven ocular lymphoma were retrieved from the files of our hospital and of a multicenter PCNSL study. In four patients, ocular lymphoma was the first manifestation of PCNSL, in three a cerebral lesion had occurred in the first place. Progression to cerebral lymphoma was seen in three of the four patients with initial eye manifestation. Imaging was performed using a dedicated thin section protocol in four patients. An intraocular abnormality was found in four cases, always in T1-weighted images after contrast injection. Differential diagnosis from uveitis or ocular melanoma was not possible by imaging alone. The examination was falsely negative in the remaining three patients.Hence, imaging has a low sensitivity for ocular lymphoma and does not facilitate differential diagnosis against uveitis or ocular melanoma. PMID- 12529796 TI - Creatine monohydrate in myotonic dystrophy: a double-blind, placebo-controlled clinical study. AB - We assessed safety and efficacy of creatine monohydrate (Cr) in myotonic dystrophy (DM1) in a double-blind, cross-over trial. Thirty-four patients with defined DM1 were randomized to receive Cr and placebo for eight weeks (10.6 g day 1-10, 5.3 g day 11-56) in one of 2 treatment sequences. There was no significant improvement using manual and quantitative muscle strength, daily-life activities, and patients' own global assessment comparing verum with placebo administration. Cr supplementation was well tolerated without clinically relevant side effects, but did not result in significant improvement of muscle strength or daily-life activities. PMID- 12529797 TI - Changes in motor cortex inhibition over time in patients with amyotrophic lateral sclerosis. AB - Abnormal balance between intracortical inhibitory and excitatory mechanisms has been found to contribute to the genesis of motor cortex hyperexcitability in amyotrophic lateral sclerosis (ALS), but data are lacking on the role of these abnormalities in the pathophysiology of the disease. We evaluated the resting motor threshold (RMT), the cortical silent period (CSP) to single-pulse transcranial magnetic stimulation (TMS), early intracortical inhibition (ICI), early intracortical facilitation (ICF) and late ICI to paired-pulse TMS in 40 patients with ALS. These parameters were correlated with disease duration and clinical features. They were also monitored over time in selected patients. The main abnormal TMS findings were: (a). reduced or even absent early and late ICI; six out of 9 patients, with normal early ICI at the first recording, developed abnormal ICI after several months; (b). reduced cortical silent period duration with increasing TMS intensity. ICF and RMT were not affected. Impairment of early and late ICI correlated significantly with disease duration, the diagnostic categories and the clinical evidence of upper motor neuron involvement. The alteration of different cortical inhibitory functions seems to take place with disease progression, rather than being the primary event in the pathogenesis of ALS. The impaired inhibition is considered as being due to both depletion of specific subpopulations of intracortical GABAergic neurons and mechanisms involved in motor cortex reorganization following progressive neuronal loss. Clarification of the importance of these factors in the pathogenesis of the disease may have diagnostic and therapeutic implications. PMID- 12529798 TI - Botulinum toxin type B for treatment of axillar hyperhidrosis. AB - Recently, botulinum toxin type B (BT-B) became commercially available for treatment of cervical dystonia. It is the aim of this study to explore its use for treatment of bilateral axillar hyperhydrosis (HH). For this we directly compared the antihyperhydrotic effect of BT-B (NeuroBloc)/MyoBloc) with that of botulinum toxin type A (BT-A) (Botox). 9 patients (HD group) received BT-A 100MU unilaterally and BT-B 4000MU contralaterally. 10 patients (LD group) received BT A 100MU and BT-B 2000MU. All patients were blinded as to which preparation was used in which side. All patients except one reported excellent HH improvement in both axillae. None of the patients had residual HH on clinical examination. The duration of HH improvement until first recurrence in the HD group was 16.0 +/-4.3 weeks in the BT-A treated axillar and 16.4 +/-4.5 weeks in the BT-B treated axillae (Wilcoxon rank-sum test, p = 0.336). In the LD group it was 16.4 +/-5.3 weeks in the BT-B treated axillae and 17.1 +/-5.7 weeks in the BT-A treated axillae (Wilcoxon rank-sum test, p = 0.059). There was also no difference in the duration of HH improvement between the axillae treated with BT-B 4000MU and BT-B 2000MU (Wilcoxon rank-sum test, p = 0.712). 5 out of 9 patients in the HD group (chi-square test, p = 0.025) and 7 out of 10 patients in the LD group (chi-square test, p = 0.008) reported more application discomfort in the BT-B treated axillae. In 6 out of 9 patients in the HD group (chi-square test, p = 0.014) and in 6 out of 10 patients in the LD group (chi-square test, p = 0.014) the onset of HH improvement appeared earlier in the BT-B treated axillae. One patient in the HD group reported dryness of the mouth and eyes and accomodation difficulties.BT B is a safe and efficient treatment for axillar HH. Doses of BT-B 2000MU per axilla seem sufficient indicating a conversion factor between BT-A and BT-B in the order of 1:20. With a conversion factor for cervical dystonia in the order of 1:40 the autonomic nervous system seems to be relatively more sensitive to BT-B than to BT-A compared with the motor system. PMID- 12529799 TI - The indication for liver transplant to improve neurological symptoms in a patient with Wilson's disease. PMID- 12529800 TI - Diffusion- and perfusion-weighted MRI demonstrates synergistic lesions in acute ischemic Foix-Chavany-Marie syndrome. PMID- 12529801 TI - Dropped head as the presenting symptom of primary hyperparathyroidism. PMID- 12529802 TI - Early and persistent sensory-psychiatric symptoms in an inherited prion disease with a PrP P105L mutation. PMID- 12529803 TI - I-123-metaiodobenzylguanidine uptake of the forearm shows dysfunction in peripheral sympathetic mediated neurovascular transmission in complex regional pain syndrome type I (CRPS I). PMID- 12529804 TI - Anterior spinal artery syndrome: the diagnostic value of electromyography (EMG). PMID- 12529805 TI - Hemichorea as a presentation of recurrent non-Hodgkin's lymphoma. PMID- 12529806 TI - Edme Felix Alfred Vulpian (1826-1887). PMID- 12529807 TI - Lorazepam substitutes for the alcohol stimulus in social drinkers. AB - RATIONALE: The alcohol discriminative stimulus has been extensively studied in animals and demonstrated to be pharmacologically complex. In contrast, however, the alcohol stimulus has been less frequently studied in humans. OBJECTIVES: The aim of the experiments reported here was to characterise pharmacologically an alcohol discriminative stimulus in social drinkers. METHODS: Volunteers were first trained to discriminate a dose of 0.2 g/kg alcohol from placebo, using an established method. We then investigated the generalisation response and subjective effects following a range of doses of the gamma-amino-butyric acid (GABA)(A) benzodiazepine-receptor agonist lorazepam (0, 0.5, 1 and 2 mg, PO). RESULTS: Low doses of lorazepam (0.5 and 1 mg) did not cross-generalise with the alcohol stimulus and produced only minimal subjective effects. However, a dose of 2 mg lorazepam substituted (60.8%) for the stimulus ( P<0.02) and increased subjective ratings of "lightheaded" ( P<0.05). CONCLUSIONS: These results are consistent with the pre-clinical literature and indicate the cross-species generality of the GABA(A) component of the alcohol discriminative stimulus. PMID- 12529809 TI - Pentagastrin test for anxiety--psychophysiology and personality. AB - RATIONALE: CCK(B) receptor agonists such as pentagastrin or CCK(4) have anxiogenic panic-like effects in humans. Our previous findings are in agreement with a relationship between C-peptide plasma levels (as a measure of insulin release) and sensitivity to psychotropic activation by CCK(B) receptor stimulation. OBJECTIVES: Our present aim was to study the transient humoral, physiological and psychotropic effects of pentagastrin bolus injection and whether personality might affect the outcome. METHODS: Pentagastrin was given in bolus i.v. injections to healthy volunteers at increasing doses (0.003, 0.012, 0.05 and 0.2 microg/kg). Physiological parameters were recorded before and during each pentagastrin challenge and humoral variables described previously were included in the analyzes. Subjects rated the discomfort following pentagastrin administration on the state anxiety and discomfort scale (SADS). The Karolinska scale of personality (KSP), anxiety sensitivity index (ASI) and Hamilton anxiety scale (HAS) were used to characterize the subjects before the test. RESULTS: Galvanic skin response (GSR) and heart rate (HR) were significantly increased within 1 min following 0.2 microg/kg pentagastrin, the GSR increase correlating negatively to the C-peptide increase and positively to ratings on SADS. Even the lower dose (0.05 microg/kg pentagastrin) induced a significant increase in GSR. ASI measures correlated weakly to the increase in ratings on SADS following 0.05 microg/kg and 0.2 microg/kg pentagastrin. CONCLUSIONS: We found correlations between psychophysiology, humoral response and subjective ratings following pentagastrin administration. CCK(B) receptor stimulation might reveal phenotype properties predictive of anxiety-related traits, measurable through serum levels of C-peptide. Furthermore, our results support the predictive value of ASI for fearfulness. PMID- 12529808 TI - Effects of fluoxetine and d-fenfluramine on cocaine-seeking behavior in rats. AB - RATIONALE: Serotonin (5-HT) systems may play a role in modulating cocaine-seeking behavior. OBJECTIVES: The present study examined the effects of acute administration of the 5-HT reuptake inhibitor (SRI) fluoxetine, and the SRI/releaser d-fenfluramine, on reinstatement of extinguished cocaine-seeking behavior elicited by either response-contingent presentations of cocaine-paired cues or cocaine priming. METHODS: Separate groups of rats that had been trained to press a lever for a cocaine reinforcer (0.75 mg/kg per 0.1 ml, IV) with a light/tone stimulus complex paired with each infusion underwent daily extinction sessions during which responding had no scheduled consequences (i.e. neither cocaine nor the stimulus complex was available). Subsequently, the effects of fluoxetine (0-10.0 mg/kg, IP) on extinction and cue reinstatement of extinguished cocaine-seeking behavior were examined, as well as the effects of d-fenfluramine (0-3.0 mg/kg, IP) on cue reinstatement. Additionally, dose-dependent effects of fluoxetine (0-10.0 mg/kg, IP) and d-fenfluramine (0-1.0 mg/kg, IP) on cocaine primed (0-15.0 mg/kg, IP) reinstatement of extinguished cocaine-seeking behavior were examined. RESULTS: Fluoxetine dose-dependently attenuated cocaine-seeking behavior during extinction. Both fluoxetine and d-fenfluramine dose-dependently attenuated cue-reinstated cocaine-seeking behavior. In contrast, neither drug reliably altered cocaine-seeking behavior reinstated by cocaine priming. CONCLUSIONS: These findings suggest that 5-HT indirect agonists effectively attenuate cocaine-seeking behavior elicited by cocaine-associated stimuli, but are much less effective in attenuating cocaine-seeking behavior elicited by cocaine priming. PMID- 12529810 TI - The ascending limb of the cocaine dose-response curve for reinforcing effect in rhesus monkeys. AB - RATIONALE: To date, the literature on the intravenous self-administration of cocaine by laboratory animals lacks a compelling demonstration of an ascending limb to the dose-response function. It has been argued that previous demonstrations of an ascending limb are confounded by the extinction process. OBJECTIVE: The objective was to examine the relationship between cocaine dose and intravenous self-injection frequency at the low end of the cocaine dose range (0.03-0.00075 mg/kg per injection). METHODS: Three adult rhesus monkeys were given the opportunity to self-inject cocaine on a fixed-ratio 1 schedule of reinforcement with no timeouts between injections. Single cocaine doses were presented for between 13 and 27 consecutive 2-h sessions in the order of 0.03, 0.01, 0.003, 0.0015(a), 0.00075, and 0.0015(b) mg/kg per injection. RESULTS: An ascending limb of the cocaine dose-response curve was found to exist between the doses of 0.00075 and 0.003 mg/kg per injection. CONCLUSIONS: The fact that response rate increased from 0.00075 to 0.0015(b) mg/kg per injection, and remained stable at this intermediate level, negates the possibility that responding at 0.0015(b) mg/kg per injection is an artifact of experimental extinction. The finding that significantly less cocaine was taken at 0.0015(b) mg/kg per injection than at higher doses demonstrates that satiety was not the mechanism by which cocaine intake was regulated on the ascending limb of the dose response curve. PMID- 12529811 TI - [Surgery for morbid obesity]. PMID- 12529812 TI - [New trends in hepato-biliary surgery]. PMID- 12529813 TI - [Development, state of the art and future perspectives of surgery for morbid obesity]. AB - Obesity is the most common form of malnutrition and morbid obesity is increasingly recognized as a major health concern. Drug treatments of the morbid obese have entirely failed in the long run. These circumstances have constituted incentives for surgeons to develop techniques resulting in malabsorptive or restrictive effects on food intake. Several techniques achieve weight loss through both these mechanisms and most likely also by changing the gastrointestinal signalling system. Within the last 50 years surgery for morbid obesity has been developed stepwise, like other surgical methods. More than 50 surgical anti-obesity techniques have been described. Half a dozen techniques have been left over and are performed mainly laparoscopically now a days. The outcome of these techniques over the last 30 years is reported in this article. It is the aim to illustrate that bariatric surgery is safe and so far more long term efficient than any other technique available for the treatment of morbid obesity. PMID- 12529814 TI - [Bariatric surgery -- stereotypes and paradigms]. AB - Many physicians regard obesity as a sin and treat fat patients with disdain befitting a moral leper. Non-bariatric physicians, being a product of our culture, seem more likely to have an obesity paradigm close to that of the public. Many members of the public regard obesity surgery as dangerous. Many insurers reject morbid obese patients from bariatric surgical treatment with the paradigmal statement that obesity is totally the fault of a fat person. These medical experts do not accept obesity as a disease (which WHO does) and therefore social courts also reject applications of patients who want to undergo bariatric surgery. Morbid obesity is a multifactorial problem with genetic, biochemical, hormonal, environmental, behavioral and cultural elements. It is recognized as an extreme health hazard which is rarely the result of an aberrant moral problem or true addictive behavior. We need to change effectively the negative paradigms towards obesity and its surgery from some of our colleagues, hospital administration, medical insurers and the public. The existing prejudices are not acceptable. PMID- 12529815 TI - [Criteria for selection of patients for bariatric surgery]. AB - Because of the high prevalence of co-morbid conditions and poor life expectancy a Body Mass Index (BMI) of 40 kg/m(2) or more is an indication for surgery in a fully informed, consenting adult in optimal medical condition to tolerate general anaesthesia. Patients with BMI of 35-40 kg/m(2) and the existence of one or more serious obesity-related conditions ameliorated by weight loss, such as hypertension, pulmonary insufficiency, non-insulin-dependent diabetes mellitus etc., are also candidates for surgical treatment. The bariatric surgeon should use these international criteria as guidelines only, not strict rules. Attempts on the part of internists and more frequently insurance carriers to require documented failure of previous non-operative treatment is not meaningful. PMID- 12529817 TI - [Malabsorption methods in bariatric surgery]. AB - In Europe, the incidence of morbid obesity amounts approximately 1% of a nation's population. At the time being, a surgical procedure represents the only effective long-term therapy of morbid obesity. In this regard, the gastric bypass is the most common surgical method in the USA, also gaining relevancy in Europe. The development, the basic principle and the actual operation technique are shown. The mode of functioning, complications, surgical alternatives, risks, results and outcome are discussed. Over the past years, bariatric procedures and techniques have been improved, which resulted in a reduction of the risk and in an improvement of the results. Long- term success can be achieved regarding weight reduction and decrease of comorbidities. The gastric bypass is a safe and effective surgical procedure in the treatment of morbid obesity. PMID- 12529816 TI - [Restrictive procedures in the treatment of morbid obesity -- vertical banded gastroplasty vs. adjustable gastric banding]. AB - Vertical banded gastroplasty (VBG) has been in clinical use since 1979 and the adjustable gastric banding (AGB) since 1985. The aim of this study was to compare the outcome, short- and long-term complications of the two procedures. Within a period of 9 years (1992-2001) 1 011 gastric restrictive procedures were performed by two surgeons in the course of a prospective nonrandomized comparative trial. 563 VBG's via laparotomy and 448 laparoscopic AGB's were included into the study. The mean BMI of the patients was 46 kg/m 2 in both groups. The mean duration of follow up was 60 months (range 6 to 108 months). No statistical significant difference in outcome in terms of weight loss, reduction of co-morbidity and improvement in quality of life following ASGB or VBG was observed. The hospital mortality rate was 0.3 % (2 VBG, 1 AGB). The overall reintervention rate for long term complications was 15.6 % for the VBG and 7 % for the AGB group (p < 0.0001). The AGB is entirely reversible and the less invasive procedure preserving an intact anatomy of the stomach. A trend in favour of the AGB is observed. PMID- 12529818 TI - [Intragastric stimulation (IGS) for treatment of morbid obesity]. AB - The technique of implantable gastric stimulator (IGS(R)) placement for treatment of morbid obesity is described. There are three components of the IGS, the programming system, the lead and the electrical stimulator itself. It is placed in the anterior abdominal wall. It is connected to the bioplar lead that is positioned in the muscle wall of the stomach. The nominal parameters were: pulse width 208 micro sec 24 hours per day, pulse rate 40 Hertz and amplitude 6.2-10.0 mA. Fourty eight patients have been treated using techniques that were developed and refined around the world in a prospective open study. 32 and 7 patients, resp. were available for investigation after 6 and 15 months. Twenty-seven (56 %) intra-operative gastric penetrations occurred, as noted on operative gastroscopy, without any clinical sequelae. Six patients (12 %) showed lead dislodgements and a new procedure for lead replacement. There were no operative deaths. All procedures were successfully completed laparoscopically. Two connections required revision (4 %) because the leads were not fully inserted into the lead connector of the generator. No abscess formation or severe complication were noted. The excessive weight loss after 15 months ranged to 32 %. The operation for IGS placement is safe and simple to perform. Attention to technical details is essential for safe performance of the procedure. PMID- 12529819 TI - [Dietetic treatment after gastro-restrictive operations]. AB - Bariatric surgery patients need a very intensive and individual nutritional consulting. It is not just enough to say: "eat puree food for a longer period of time". The capacity of the stomach has changed but not the desire to eat; e. g. high caloric food and also not automatically the eating behaviour. With a guideline to nutritional therapy step by step the consultations may be structured very efficiently and the possibility to reach the goal increases. The results of a survey in 150 patients are demonstrated. PMID- 12529820 TI - [Psychological aspects of bariatric surgery]. AB - Experience with gastro-restrictive obesity showed the necessity of detailed research in three main tasks: 1. characteristics of morbidly obese people 2. psychological effects of surgery 3. interaction of these characteristics and effects of surgery. We carried out 7 studies with a total of 650 participants. Clinical interviews and tests measuring personality, eating behaviour, self esteem, addiction factors and quality of life were performed. Morbidly obese differ significantly from normal weight people with the exception of most personality scales. Effects of surgery can be summarized that patients learn to avoid overeating but they do not learn to nourish on healthy solid nutrition and they do not learn to exercise more than before. By paradox learning process 30-50 % of gastric banding patients establish vomiting behaviour or eating pulp and sweets. These who show low self esteem, high addiction score and high disinhibition behaviour are more at risk. Different psychological, nutritional as well as physiotherapeutic interventions are needed to help these patients. PMID- 12529821 TI - [The Body-Mass-Index (BMI) has no impact on the frequency of typical reflux symptoms -- results of a nationwide telephone-based informing campaign in Germany]. AB - BACKGROUND AND AIM: The effects of obesity on the gastrooesophageal reflux disease (GERD) are controversial. The aim of the study was to assess the relationship between the BMI and the frequency of reflux symptoms in a population with typical GERD symptoms. METHODS: Based upon a nationwide informing campaign up to 5,000 subjects contacted the informing calling center. Subjects were included if they had heartburn and acid regurgitation. Age, gender, height, weight and the frequency of reflux symptoms were assessed by telephone interviewing. RESULTS: 1,296 subjects (668 female) with mean age of 54 +/- 14 years and a mean BMI of 26 +/- 4 were included in the study. 41.2% of the subjects had a BMI up to 25, 41.4 % between 25 to 30, and 13 % greater than 30. 74.5 % of all subjects reported reflux episodes daily and several times a week. 74.6 % of the subjects had reflux symptoms for more than one year. Both the frequency and pattern of reflux symptoms did not differ significantly in the three BMI-classes (p > 0.05, table). CONCLUSION: In the present population with typical and frequent GERD symptoms the BMI showed no impact on the frequency of reflux symptoms. A high BMI does not appear to be a risk indicator for GERD. Interventional studies are needed to assess whether a high BMI is also no risk factor for GERD. PMID- 12529822 TI - [Is duodeno-gastro-esophageal reflux just a bystander of acid reflux?]. AB - BACKGROUND & AIM: Duodeno-gastro-esophageal reflux (DGER) as measured with bilirubin monitoring is observed in many patients with reflux disease especially in Barrett's esophagus. As acid suppression is an effective therapy of reflux disease, DGER is frequently just considered as a bystander of acid reflux. To define the importance of DGER, reflux of duodenal contents was evaluated by bilirubin monitoring in the stomach. METHODS: 100 patients with reflux disease were evaluated (62 m, 38 f, 50 (12) years). 26 patients had Barrett's esophagus, 57 had esophagitis and 17 non- erosive reflux disease (pH pos.). All patients were evaluated with simultaneous 24-hour bilirubin monitoring in the esophagus and stomach. Results were compared to 35 measurements of healthy volunteers in the esophagus and 41 measurements in the stomach. RESULTS: Normal values: DGER - Exposure time esophagus > 11.8 % using an absorbance value > 0.14, duodenogastric reflux (DGR) - Exposure time stomach >24.8 % using an absorbance value > 0.25. 56 % of the patients had DGER, 41 % had DGR. 29 of the 41 patients with DGR had DGER (71 %), while 27 of the 56 patients with DGER had physiologic duodenogastric reflux (48 %). DISCUSSION: About 30 % of the patients with reflux disease show DGER combined with excessive DGR. Therefore, DGER is not just a bystander of acid reflux. The excessive DGR in some patients adds additional potentially dangerous substances to the esophageal reflux. PMID- 12529823 TI - [Risk estimation in Barrett's esophagus: biomolecular marker and histopathologic classification]. AB - The diagnosis of Barrett's esophagus and the different degrees of intraepithelial neoplasia appear to be demanding in several aspects. Current data on genetic alterations involved in the carcinogenesis of Barrett's esophagus are discussed. Several new biomarkers are being tested to help better to determine the risk of cancer development. However, a few immunohistochemical markers have emerged which could be helpful for the differential diagnosis of low- and high-grade intraepithelial neoplasia. Markers which could predict the progression of premalignant Barrett's epithelium to carcinoma are still to be established. At present, the "gold standard" for classifying the malignant potential in Barrett's esophagus is the degree of intraepithelial neoplasia found on standard biopsy protocols. PMID- 12529824 TI - [Surgery of complicated gastroduodenal ulcers: outcome at the millennium]. AB - INTRODUCTION: Complications of gastroduodenal ulcers such as bleeding and perforation have been afflicted with a mortality of up to 40 % in the past. Altered ulcer manifestation by improvement of medical treatment as well as advances in surgical techniques and intensive care medicine might currently have improved the outcome of these severely ill patients. PATIENTS AND METHODS: A retrospective analysis of 73 patients with emergency operation for bleeding or perforated gastric (n = 50) and duodenal ulcer (n = 23) between 10/1994 and 7/2001 was performed. RESULTS: The median age was 58 years (17-90 years) with 30 % of patients exceeding the age of 70 years. Eighty-one percent of patients had perforation, 19 % had bleeding. Factors predisposing for gastroduodenal ulcers were present in 88 %. Five percent of gastric perforations were due to gastric malignancies. Diagnosis of perforation was made in 88 % by conventional X-rays, in 12 % of perforations no free abdominal gas was detectable. A local surgical procedure (excision of ulcer and sutures/pyloroplasty) was performed in 64/73 patients (88 %), partial gastroduodenal resection was necessary in 9 patients (12 %). Histologic examination for helicobacter pylori was positive in 30/39 specimens (77 %). In-hospital mortality was 14 %, both for patients with bleeding and perforated ulcers. Surgical complications were 12 %. DISCUSSION: Altered ulcer manifestation, minimized surgical therapy and improved intensive care medicine led to a recent reduction in postoperative mortality of patients suffering from perforated or bleeding gastroduodenal ulcer. A further reduction of this still high mortality might be expected by improvement of surgical training and/or restriction of therapy to a limited number of surgeons. PMID- 12529825 TI - [The medical advisory services of social health insurance]. AB - A social health insurance needs opinions on diagnostic and therapeutic procedures - for the single question regarding the individual patient and as general assessments (health technology assessments) as well. The Medical advisory services of social health insurance (their federal branches and their central board) have to do more than 9 Mio individual expert opinions and more than 100 general assessments or HTA-reports a year. Several examples are to be shown. PMID- 12529826 TI - [Intragastric penetration of a gastric band -- a severe complication after gastric banding?]. AB - In morbid obese patients the adjustable laparoscopic gastric banding is an effective procedure with a widespread acceptance. Pouch dilatation, band slipping, port-penetration and port infection are the most known complications. We report on three patients with a penetration of an adjustable gastric band. The main symptom of penetration was the increase of weight. The reasons for penetration are a lesion of the serosa intraoperatively or a overfilling of the band. In our patients the treatment was in two cases the explantation of the band. In one case the patient showed no signs of peritonitis and didn't want the explantation of the band. Our recommendation in case of penetration is the removal of the band, because every penetration can be the source of a lethal outcome. PMID- 12529829 TI - [Cognitive psychology and clinical neuroscience]. PMID- 12529827 TI - [Duodenal ulcer presenting as pneumomediastinum and pneumothorax -- case report]. AB - Haemorrhage, penetration and perforation are common complications of peptic ulcers. Free intraabdominal air is seen in 80 % after perforation. Penetration into the retroperitoneum with pneumothorax and mediastinal emphysema are rarely observed. We report the case of a 85-year-old female patient with nausea, vomiting and little appetite. During endoscopy of the upper GI-tract she complained about progressive dyspnea. Chest X-ray revealed mediastinal emphysema and pneumothorax. When performing laparotomy, we found a duodenal ulcer, that had penetrated the retroperitoneal space. The patient underwent partial gastrectomy and reconstruction with Billroth-II anastomosis. The postoperative course was uneventful. PMID- 12529830 TI - [Neglect dyslexia owing to traumatic fronto-temporal right hemisphere bleeding]. AB - Apart from unilateral disturbance of the egocentric reference system, as it is traditionally known of neglect, there are also cases with unilateral disturbances of a stimulus-centered or object-centered reference system. In these cases the spatial position of the object, from the observer's perspective, plays only a limited role in the explanation of the problems with object perception. We describe a patient who, following head-brain trauma and a fronto-temporal bleeding, showed, beside traditional neglect, also a stimulus-centered left-sided disorder in word and object perception. Our investigations showed that this disorder (a) is independent of the position of the object in the environmental space, (b) comprises words and objects, and (c) is especially sensitive for changes in the first letters of a word. Conclusively, it yields from this case that at least three different reference systems of object representation should be distinguished in neglect, which can be disturbed independently from each other and may negatively influence the behavioral potential of the patients. PMID- 12529831 TI - [Trigemino-autonomic headache syndromes]. AB - This review describes the characteristics of some rare severe primary headache syndromes without any structural lesions, which are summarized as the trigemino autonomic headaches. Pain in these syndromes is associated with autonomic symptoms such as conjunctival injection, lacrimation, rhinorrhoe, ptosis or eyelid edema. This article summarizes the diagnostics, epidemiology and therapy of these conditions. PMID- 12529832 TI - [The HPA-axis as a possible link between depression, diabetes mellitus and cognitive dysfunction]. AB - Patients suffering from diabetes mellitus and depressive patients show a hyperactivity of the hypothalamic-pituitary-adrenocortical axis (HPA-axis) with hypercortisolemia. Hypercortisolemia is associated with cognitive dysfunction. These neuroendocrinological disturbances can cause an insulin resistance syndrome which complicates the regulation of blood glucose. Cognitive and depressive disorders in patients with diabetes mellitus might be associated with a hyperactivity of the HPA-axis. By normalising the HPA-axis both disorders could be improved. In addition, one can expect that antidepressive treatment with normalization of the HPA-axis could improve the metabolic situation and cognitive dysfunction. There is need for further research to study the associations between depression, diabetes mellitus and cognitive dysfunction. PMID- 12529833 TI - [Psychiatric comorbidity in risk groups of opioid addiction: a comparison between opioid dependent and non-opioid dependent prisoners (in jail due to the German narcotics law)]. AB - Individuals who do not develop opioid dependence although they have access to opioids might differ with regard to psychiatric risk factors from opioid-exposed subjects who developed opioid dependence. To test this assumption, the present investigation compared individuals who were in jail due to the German "Dangerous Drugs Act" (i. e. particular risk group due to facilitated opioid availability) according to presence or absence of opioid dependence and psychiatric comorbidity in each group. This study design is in line with the assumption that in addition to the (postulated) environmental risk factor of facilitated availability of opioids, psychiatric risk factors enhance the likelihood for the development of opioid dependence. Opioid addicts represent a risk group, not only for other forms of substance dependence, but also psychiatric disorders like anxiety disorders, suicide attempts and specific forms of personality disorders. However the difference between opioid dependent subjects and non-opioid dependent controls was less marked than initially postulated. Alcoholism of non-opioid dependent prisoners also was associated with depressive episodes, anxiety disorders as well as cocaine dependence. Despite the high frequency of life-time psychiatric comorbidity in the opioid dependent sample, this increased comorbidity was not paralleled by psychiatric treatment. In general, the sample of prisoners investigated here, was characterized by a high frequency of psychiatric disorders including substance dependence. PMID- 12529834 TI - [Treatment of hysteria in the 19th century--in which way did German psychiatrists view castration?]. AB - Until the late 19 th century treatment of hysteria, this inhomogeneous group of somatic, neurological and psychiatric symptoms, hardly differed from the methods of the Greco-Roman epoch. Dietetics, physical applications, and smaller surgical operations made up the standard forms of intervention for this illness which was regarded as belonging to the field of gynecology. Thus, it is not astonishing that oophorectomy (ovariectomy) was included into the therapeutic spectrum for hysteria both in Germany and beyond at the end of the 19 th century. It was above all gynecologist Alfred Hegar (1830 - 1914) who tried to extend the application of this method to nervous illnesses. This therapeutic alternative did not, however, meet with an unanimously positive response among psychiatrists. Richard von Krafft-Ebing (1840 - 1902) as a follower of the degeneration theory as well as Paul Flechsig (1847 - 1929) supported it, applying it as the ultima ratio. But since these operations produced no lasting curative impact, functional neurological disorders were regarded more important. From Emil Kraepelin (1856 - 1926) then, psychiatrists began to understand the psychological component of this illness, for which Leipzig neurologist Paul Julius Mobius (1853 - 1907) had made major contributions. Thus, surgical interventions were rejected and conservative methods of treatment were developed further considering psychological aspects. It was Adolf Strumpell (1853 - 1925) who had laid the basis for this postulating a "psychic trauma" as the cause for hysteria as early as in 1884. On the other hand, Mobius had developed a psychotherapeutic concept for the treatment of these disorders long before Sigmund Freud (1856 - 1939) came out with his psychocathartic method. PMID- 12529835 TI - [Treatment of obstructive sleep apnea with a new vest preventing the supine position]. AB - BACKGROUND AND OBJECTIVE: Prevention of the supine position has for decades been a well known treatment option in positional obstructive sleep apnea. It was the aim of this study to test the efficacy of a recently patented commercially available supine position preventing vest. PATIENTS AND METHODS: The vest is made of linen tissue with a half cylindrical piece of hard foam in its dorsal part. 12 male patients slightly overweight (body mass index 26.5 +/- 2.6 kg/m(2)) and 55.8 +/- 11.6 years old were investigated polysomnographically prospectively. RESULTS: The pretherapeutical respiratory-disturbance index (RDI) was 26.7+/-11.9/h increasing to 39.3+/-16.1/h in the supine position. When using the vest, patients no longer slept on their back and the RDI dropped to 7.6 +/- 5.1/h (p < 0.005). Total sleep time at an oxygen saturation below 90 % was reduced from 11.7 +/- 11.3 % to 1.5 +/- 2.1 %. 9 patients (75 %) were cured (RDI < 10/h and RDI reduction > 50 %), 2 patients (17 %) improved (RDI reduction > 50 %) and only the oldest patient (76 years old) remained unchanged. Even though snoring decreased from 180 +/- 125 minutes to 110 +/- 52 minutes, an increase was observed in 30 % of the patients. Sleep quality and structure did not change considerably. CONCLUSION: The supine position prevention vest is a safe and simple treatment alternative of high efficacy in positional sleep apnea. As a complete remission cannot be predicted, polygraphic or polysomnographic controls are necessary. PMID- 12529836 TI - [Validity of iohexol clearance in patients with chronic renal failure and normal renal function in comparison to (99m)Tc-DTPA-clearance]. AB - BACKGROUND AND OBJECTIVE: The most common method used for testing dynamic renal function is creatinine clearance, but it has some limitations, e. g. variable muscle mass and tubular secretion of creatinine. The use of radionuclides as an exact method is limited in terms of availability, cost and time needed for examination. We compared the plasma clearance of iohexol with the established (99m)Tc-diethylenetriaminepentaacetate acid (DTPA) clearance. The aim of the present study was to validate iohexol clearance as a simple and suitable method for measuring to determine GFR with a comparable sensitivity to radioisotopic methods. METHODS: 120 patients (49 females, 71 males), mean age of 56 (range 20 to 84) years with normal renal function and different stages of renal failure, mean creatinine clearance of 61.6 +/- 44,9 (range 1.8 - 181.1) ml/min/1.73 m2 received a bolus injection of 10 ml iohexol, a non-ionic low osmolar x-ray contrast medium. Using the one-compartment model, plasma samples were taken after 150, 240 and 480 minutes. The total plasma disappearance of iohexol was measured by x-ray fluorescence analysis and the clearance was calculated. The (99m)Tc-DTPA clearance was determined in accordance with a standard protocol. RESULTS: A high correlation was found between the clearance of iohexol and (99m)Tc-DTPA (r = 0.95). The average deviation between Iohexol and (99m)Tc-DTPA clearance was 7.4 ml/min/1.73 m2. Allergic and nephrotoxic side effects were not observed. CONCLUSION: Iohexol clearance is a valid method for measuring GFR in patients at any stages of renal failure. It is easy to perform and inexpensive. PMID- 12529837 TI - [Fatty liver and increased serum lactate in a woman with HIV]. AB - HISTORY AND CLINICAL PRESENTATION: A 50-year-old woman with HIV who had undergone a 7-month course of antiretroviral treatment (stavudine, lamivudine, indinavir, ritonavir) was admitted to hospital with abdominal pain, nausea, vomiting and muscle pain. Physical examination revealed subfebrile body temperature and hepatomegaly. INVESTIGATIONS: Biochemical tests revealed moderate elevations of gamma GT, AST and ALT as well as marked hyperlactatemia. Abdominal computed tomography indicated pronounced hepatomegaly. Percutaneous liver biopsy was performed and demonstrated macro- and microvesicular hepatic steatosis on light microscopy. Electron microscopy showed mitochondrial damage. TREATMENT AND COURSE: Despite discontinuation of antiretroviral treatment serum lactate levels further increased initially and clinical symptoms were progressive. In the following weeks, a slow but steady recovery of laboratory values and physical condition was noted. CONCLUSION: Hyperlactataemia associated with abdominal pain can indicate the mitochondrial toxicity due to nucleoside reverse transcriptase inhibitors. After drug discontinuation, improvement of clinical symptoms and laboratory values is often delayed, frequently for several weeks. PMID- 12529838 TI - [Microsporidiosis in humans: parasitology, clinical features and treatment]. PMID- 12529839 TI - [Endothelial function of the human renal vasculature]. PMID- 12529840 TI - [Diagnosis of coronary heart disease in women. Peculiarities, difficulties and risk stratified management]. PMID- 12529841 TI - [Providing or withholding treatment:--factors influencing clinical decision making--the role of the patient's wish and priorities in palliative care]. PMID- 12529842 TI - ["Shared decision making": doctor or patient--who decides?]. PMID- 12529843 TI - Metabolism, swimming performance, and tissue biochemistry of high desert redband trout (Oncorhynchus mykiss ssp.): evidence for phenotypic differences in physiological function. AB - Redband trout (Oncorhynchus mykiss ssp.) in southeastern Oregon inhabit high elevation streams that exhibit extreme variability in seasonal flow and diel water temperature. Given the strong influence and potential limitations exerted by temperature on fish physiology, we were interested in how acute temperature change and thermal history influenced the physiological capabilities and biochemical characteristics of these trout. To this end, we studied wild redband trout inhabiting two streams with different thermal profiles by measuring (1) critical swimming speed (U(crit)) and oxygen consumption in the field at 12 degrees and 24 degrees C; (2) biochemical indices of energy metabolism in the heart, axial white skeletal muscle, and blood; and (3) temperature preference in a laboratory thermal gradient. Further, we also examined genetic and morphological characteristics of fish from these two streams. At 12 degrees C, maximum metabolic rate (Mo2max) and metabolic power were greater in Little Blitzen redband trout as compared with those from Bridge Creek (by 37% and 32%, respectively). Conversely, Bridge Creek and Little Blitzen trout had similar values for Mo2max and metabolic power at 24 degrees C. The U(crit) of Little Blitzen trout was similar at the two temperatures (61+/-3 vs. 57+/-4 cm s(-1)). However, the U(crit) for Bridge Creek trout increased from 62+/-3 cm s(-1) to 75+/-3 cm s(-1) when water temperature was raised from 12 degrees to 24 degrees C, and the U(crit) value at 24 degrees C was significantly greater than for Little Blitzen fish. Cost of transport was lower for Bridge Creek trout at both 12 degrees and 24 degrees C, indicating that these trout swim more efficiently than those from the Little Blitzen. Possible explanations for the greater metabolic power of Little Blitzen redband trout at 12 degrees C include increased relative ventricular mass (27%) and an elevation in epaxial white muscle citrate synthase activity (by 72%). Bridge Creek trout had 50% higher lactate dehydrogenase activity in white muscle and presumably a greater potential for anaerobic metabolism. Both populations exhibited a preferred temperature of approximately 13 degrees C and identical mitochondrial haplotypes and p53 gene allele frequencies. However, Bridge Creek trout had a more robust body form, with a relatively larger head and a deeper body and caudal peduncle. In summary, despite the short distance ( approximately 10 km) and genotypic similarity between study streams, our results indicate that phenotypic reorganization of anatomical characteristics, swimming ability at environmentally pertinent temperatures and white axial muscle ATP-producing pathways occurs in redband trout. PMID- 12529844 TI - The physiology of overwintering in a turtle that occupies multiple habitats, the common snapping turtle (Chelydra serpentina). AB - Common snapping turtles, Chelydra serpentina (Linnaeus), were submerged in anoxic and normoxic water at 3 degrees C. Periodic blood samples were taken, and PO(2), PCO(2), pH, [Na(+)], [K(+)], [Cl(-)], total Ca, total Mg, [lactate], [glucose], hematocrit, and osmolality were measured; weight gain was determined; and plasma [HCO(3)(-)] was calculated. Submergence in normoxic water caused a decrease in PCO(2) from 10.8 to 6.9 mmHg after 125 d, partially compensating a slight increase in lactate and allowing the turtles to maintain a constant pH. Submergence in anoxic water caused a rapid increase in lactate from 1.8 to 168.1 mmol/L after 100 d. Associated with the increased lactate were decreases in pH from 8.057 to 7.132 and in [HCO(3)(-)] from 51.5 to 4.9 mmol/L and increases in total Ca from 2.0 to 36.6 mmol/L, in total Mg from 1.8 to 12.1 mmol/L, and in [K(+)] from 3.08 to 8.45 mmol/L. We suggest that C. serpentina is tolerant of anoxic submergence and therefore is able to exploit habitats unavailable to some other species in northern latitudes. PMID- 12529845 TI - Metabolic and thermal physiology of pigeons and doves. AB - Pigeons and doves (Columbidae) are an interesting group to examine for physiological adaptations to climate and diet because this cosmopolitan family comprises more than 300 species that are mostly granivores, although some are specialized frugivores. We determined allometric and phylogenetic effects on body temperature (T(b)), basal metabolic rate (BMR; J h(-1)), and wet thermal conductance (C(wet); J h(-1) C(-1)), and we examined mass (M) and phylogenetically corrected residuals for further effects of climate, diet, and landmass size (mainland or island). Independent contrasts, correlograms, autoregression, and phylogenetic eigenvector regression (PVR) were used to examine phylogenetically related effects. We found a small but significant phylogenetic pattern for body mass of columbids. For T(b), there was no significant effect of mass or phylogeny. There was a significant effect of climate on T(b) and no significant effects of diet or landmass without mass or phylogenetic correction, but after mass and phylogenetic correction, there were no effects of climate, diet, or landmass. For BMR, there was a strong allometric effect, and residuals were significantly lower for arid and tropical species but not for temperate species, compared to predictions for nonpasserine birds. There was a nearly significant autoregressive phylogenetic relationship for BMR parl0;r=0.44), and the strong allometry of BMR remained for independent contrasts (slope=0.731), autoregressive residuals (0.698), and PVR (0.705). Residuals, from regression of autoregression and PVR residuals of M and BMR, were significantly associated with climate: arid pigeons had a lower BMR residual than tropical and temperate pigeons. PVR residuals were significantly affected by landmass (island columbids had a smaller residual than mainland columbids), but autoregression residuals were not. There was no association of autoregression or PVR residuals with diet. For C(wet), there was a strong allometric effect, and residuals for columbids were significantly higher compared to other birds. There was no significant relationship for C(wet) of columbids to climate, diet, or landmass. There was no significant autoregressive or PVR relationship for C(wet), and the strong allometry remained after phylogenetic analysis by independent contrasts (slope=0.501), autoregression (0.509), and PVR (0.514). Residuals from autoregression and PVR were not significantly correlated with climate, diet, or landmass (mainland/island). PMID- 12529846 TI - Factors that influence assimilation rates and fractionation of nitrogen and carbon stable isotopes in avian blood and feathers. AB - By switching great skuas Catharacta skua from one isotopically distinct diet to another, we measured diet-tissue discrimination factors and tested the assumption that dietary nitrogen and carbon isotope signatures are incorporated into blood and feathers at similar rates. We also examined the effects of metabolic rate and looked for evidence of isotopic routing. We found that blood delta(15)N and delta(13)C signatures altered after the diet switch at similar rates (14.4 d and 15.7 d, respectively). Qualitative analyses imply that the same was true with feathers. Mass balance calculations suggest that only a small amount of lipid is likely to be involved in the synthesis of blood and feathers. Differences in diet tissue discrimination factors before and after the diet switch may mean that toward the end of the experiment, some of the nutrients for blood synthesis had been coming from stores. Repeated measures mixed models provided evidence that increases in metabolic rate might accelerate fractional turnover rates in blood. There is a need for more laboratory-based experimental isotope studies in order to address further questions that this study has raised. PMID- 12529847 TI - Excretory nitrogen metabolism in the juvenile axolotl Ambystoma mexicanum: differences in aquatic and terrestrial environments. AB - The fully grown but nonmetamorphosed (juvenile) axolotl Ambystoma mexicanum was ureogenic and primarily ureotelic in water. A complete ornithine-urea cycle (OUC) was present in the liver. Aerial exposure impeded urea (but not ammonia) excretion, leading to a decrease in the percentage of nitrogen excreted as urea in the first 24 h. However, urea and not ammonia accumulated in the muscle, liver, and plasma during aerial exposure. By 48 h, the rate of urea excretion recovered fully, probably due to the greater urea concentration gradient in the kidney. It is generally accepted that an increase in carbamoyl phosphate synthetase activity is especially critical in the developmental transition from ammonotelism to ureotelism in the amphibian. Results from this study indicate that such a transition in A. mexicanum would have occurred before migration to land. Aerial exposure for 72 h exhibited no significant effect on carbamoyl phosphate synthetase-I activity or that of other OUC enzymes (with the exception of ornithine transcarbamoylase) from the liver of the juvenile A. mexicanum. This supports our hypothesis that the capacities of OUC enzymes present in the liver of the aquatic juvenile axolotl were adequate to prepare it for its invasion of the terrestrial environment. The high OUC capacity was further supported by the capability of the juvenile A. mexicanum to survive in 10 mM NH(4)Cl without accumulating amino acids in its body. The majority of the accumulating endogenous and exogenous ammonia was detoxified to urea, which led to a greater than twofold increase in urea levels in the muscle, liver, and plasma and a significant increase in urea excretion by hour 96. Hence, it can be concluded that the juvenile axolotl acquired ureotelism while submerged in water, and its hepatic capacity of urea synthesis was more than adequate to handle the toxicity of endogenous ammonia during migration to land. PMID- 12529848 TI - Bioenergetics of reproduction and pup development in a subterranean rodent (Ctenomys talarum). AB - The objective of this study was to evaluate the maternal costs of reproduction and pup development in the subterranean rodent Ctenomys talarum (Thomas 1898). Statistical differences were detected in whole-animal metabolic rates between nonreproductive and pregnant or lactating females. Whole-animal metabolic rates during pregnancy and lactation were 128% and 151% of the resting metabolic rate (RMR) observed in nonreproductive females. The total additional energy cost of reproduction (above the nonreproductive level) was similar for both the gestation and lactation periods. Mass-specific RMR revealed an upregulation of cell or tissue metabolism during lactation but not during gestation. The mass-specific metabolic rate of pups was 237% of the adults' metabolic rates. No differences were observed in body temperature among nonreproductive, pregnant, or lactating females. No differences were detected in body mass at birth among pups from litters with different numbers of nestlings. Pups increased their body temperature, reaching adult temperature at 30 d of age, when they were near weaning. Milk constituted the exclusive food for pups until they started eating solid food at 10 d old. Suckling time decreased with age of pups, and at the same time, mother chases directed toward their pups increased. These reproductive characteristics may contribute to successful existence in a subterranean habitat. PMID- 12529849 TI - Energy expenditure and water flux of Ruppell's foxes in Saudi Arabia. AB - Scattered populations of Ruppell's foxes (Vulpes rueppelli) occur across the deserts of northern Africa and Arabia. Little is known about the biology of these canids, especially the physiological mechanisms that contribute to their ability to live in such harsh environments. For individuals from Saudi Arabia, we tested the hypotheses that Ruppell's foxes have a reduced basal metabolic rate and total evaporative water loss (TEWL), parameters measured in the laboratory, and a reduced field metabolic rate (FMR) and water flux when free-living. Under basal conditions in the laboratory, males, which averaged 1,858 g in body mass, had an oxygen consumption of 914.9 mL O(2)/h, whereas females, which weighed on average 1,233 g, consumed 682.9 mL O(2)/h; rates of oxygen consumption translated to 441.4 kJ/d and 329.4 kJ/d, respectively. TEWL averaged 52.6 g H(2)O/d for males and 47.5 g H(2)O/d for females. We found no evidence that basal metabolism is reduced in Ruppell's foxes, but their TEWL was remarkably low: 50.9% of allometric prediction for males and 64.5% for females. In the wild during winter, males expended energy at a rate of 1,306.5 kJ/d, whereas females had an expenditure of 722.8 kJ/d. Analysis of covariance with FMR as the dependent variable, sex as a fixed factor, and body mass as a covariate showed no statistical difference in FMR between sexes. Water flux did not differ significantly between sexes and averaged 123 mL H(2)O/d, a value 30% lower than the kit fox from the deserts of southwestern North America. FMR was positively related to nocturnal activity levels as FMR (kJ/d) = -2,900.1+55.5 (% of time moving). The water content of prey items varied between 1.9 and 4.1 g H(2)O/g dry matter consumed. Based on these values and knowledge of their diet, we calculated that foxes captured about one rodent and a variety of anthropods per night of foraging. PMID- 12529850 TI - Noninvasive estimation of body composition in small mammals: a comparison of conductive and morphometric techniques. AB - Body fat stores may serve as an index of condition in mammals. Thus, techniques that measure fat content accurately are important for assessing the ecological correlates of condition in mammal populations. We compared the ability of two conductive techniques, bioelectrical impedance analysis (BIA) and total body electrical conductivity (TOBEC), to predict body composition with that of morphometric methods in three small mammal species: red squirrels (n=13), snowshoe hares (n=30), and yellow-bellied marmots (n=4). Animals were livetrapped in northern Idaho; BIA (all subjects) and TOBEC (squirrels only) measurements were taken following chemical immobilization in the field, and morphometric measurements were taken postmortem. Information provided by BIA and TOBEC failed to improve upon the predictive power of morphometric equations for total body water (TBW) and lean body mass (LBM) in squirrels and hares, which do not store substantial amounts of fat (<5% body mass comprised of fat). Although the same pattern held with respect to LBM in marmots, which accumulate substantial amounts of body fat (>10% body mass), a BIA-based model proved best at estimating TBW, suggesting that the usefulness of conductive techniques may be a function of fat deposition. However, regardless of the technique used to predict body composition, estimates of body fat furnished by our equations failed to approximate actual fat levels accurately in all three test species, probably because these techniques only provide indirect estimates of fat content. These results highlight the limitations inherent in contemporary methods of animal fat estimation and underscore the need for the development of direct and accurate measures of body fat in mammals. PMID- 12529851 TI - Changes in blubber distribution and morphology associated with starvation in the harbor porpoise (Phocoena phocoena): evidence for regional differences in blubber structure and function. AB - To examine patterns of blubber loss accompanying a decline in body condition, blubber thickness of juvenile harbor porpoises in normal/robust body condition (n=69) was compared with that of starved conspecifics (n=31). Blubber thickness in the thorax of starved porpoises (9-11 mm) was only 50%-60% of that of normal animals (18-20 mm); however, very little tailstock blubber was lost during starvation. Adipocytes in thorax and tailstock blubber were measured in both groups (n=5) to determine whether thickness changes were homogeneous throughout blubber depth. In the thorax of normal porpoises, adipocytes near the epidermis (outer blubber) were smaller (0.11 nL) than inner blubber adipocytes (0.17 nL). Conversely, the size of tailstock adipocytes was uniform. Starved animals had fewer, smaller adipocytes in the inner thorax blubber, suggesting a possible combination of adipocyte shrinkage and loss. Lipids were withdrawn only from the inner layer of thorax blubber during starvation, supporting a hypothesis of regional specialization of function in blubber. Blubber of the thorax serves as the site of lipid deposition and mobilization, while the tailstock is metabolically inert and likely important in locomotion and streamlining. Therefore, some proportion of the blubber of small odontocetes must be considered structural/mechanical rather than an energy reserve. PMID- 12529852 TI - Thermoregulation of weaned northern elephant seal (Mirounga angustirostris) pups in air and water. AB - Fasting weaned northern elephant seal pups (Mirounga angustirostris) experience diverse environmental conditions on land and in water on a daily basis. Each environment undoubtedly induces distinct energetic costs that may vary for pups of differing body condition. To determine the energetic costs associated with different environmental conditions and whether costs vary between individuals, body mass, surface area, volume, body composition, resting metabolic rate, and core body temperature were determined for 17 weaned northern elephant seal pups from Ano Nuevo, California. Metabolic rate and body temperature were measured for pups resting in air (20.9 degrees +/-0.8 degrees C), cold water (3.8 degrees+/ 0.4 degrees ;C), and warm water (14.5 degrees+/-0.2 degrees C). Resting metabolic rate increased with body mass (range: 62.0-108.0 kg) and was also correlated with lean mass and lipid mass. Metabolic rates ranged from 293.6 to 512.7 mL O(2) min( 1) and were lowest for pups resting in cold water. Thermal conductance, calculated from metabolic rate and core body temperature, ranged from 3.1 to 15.2 W degrees C(-1), with the highest values in air and the lowest values in cold water. Metabolic responses to the three environmental conditions did not differ with individual variation in body condition. For all elephant seal pups, a consequence of high lipid content is that thermoregulatory costs are greatest on land and lowest in cold water, a pattern that contrasts markedly with terrestrial mammals. PMID- 12529853 TI - Mutations in PRKCSH cause isolated autosomal dominant polycystic liver disease. AB - Autosomal dominant polycystic liver disease (ADPLD) is a distinct clinical and genetic entity that can occur independently from autosomal dominant polycystic kidney disease (ADPKD). We previously studied two large kindreds and reported localization of a gene for ADPLD to an approximately 8-Mb region, flanked by markers D19S586/D19S583 and D19S593/D19S579, on chromosome 19p13.2-13.1. Expansion of these kindreds and identification of an additional family allowed us to define flanking markers CA267 and CA048 in an approximately 3-Mb region containing >70 candidate genes. We used a combination of denaturing high performance liquid chromatography (DHPLC) heteroduplex analysis and direct sequencing to screen a panel of 15 unrelated affected individuals for mutations in genes from this interval. We found sequence variations in a known gene, PRKCSH, that were not observed in control individuals, that segregated with the disease haplotype, and that were predicted to be chain-terminating mutations. In contrast to PKD1, PKD2, and PKHD1, PRKCSH encodes a previously described human protein termed "protein kinase C substrate 80K-H" or "noncatalytic beta-subunit of glucosidase II." This protein is highly conserved, is expressed in all tissues tested, and contains a leader sequence, an LDLa domain, two EF-hand domains, and a conserved C-terminal HDEL sequence. Its function may be dependent on calcium binding, and its putative actions include the regulation of N-glycosylation of proteins and signal transduction via fibroblast growth-factor receptor. In light of the focal nature of liver cysts in ADPLD, the apparent loss-of-function mutations in PRKCSH, and the two-hit mechanism operational in dominant polycystic kidney disease, ADPLD may also occur by a two-hit mechanism. PMID- 12529854 TI - Expansion of the fragile X CGG repeat in females with premutation or intermediate alleles. AB - The CGG repeat in the 5' untranslated region of the fragile X mental retardation 1 gene (FMR1) exhibits remarkable instability upon transmission from mothers with premutation alleles. A collaboration of 13 laboratories in eight countries was established to examine four issues concerning FMR1 CGG-repeat instability among females with premutation (approximately 55-200 repeats) and intermediate (approximately 46-60 repeats) alleles. Our central findings were as follows: (1) The smallest premutation alleles that expanded to a full mutation (>200 repeats) in one generation contained 59 repeats; sequence analysis of the 59-repeat alleles from these two females revealed no AGG interruptions within the FMR1 CGG repeat. (2) When we corrected for ascertainment and recalculated the risks of expansion to a full mutation, we found that the risks for premutation alleles with <100 repeats were lower than those previously published. (3) When we examined the possible influence of sex of offspring on transmission of a full mutation-by analysis of 567 prenatal fragile X studies of 448 mothers with premutation and full-mutation alleles-we found no significant differences in the proportion of full-mutation alleles in male or female fetuses. (4) When we examined 136 transmissions of intermediate alleles from 92 mothers with no family history of fragile X, we found that, in contrast to the instability observed in families with fragile X, most (99/136 [72.8%]) transmissions of intermediate alleles were stable. The unstable transmissions (37/136 [27.2%]) in these families included both expansions and contractions in repeat size. The instability increased with the larger intermediate alleles (19% for 49-54 repeats, 30.9% for 55-59, and 80% for 60-65 repeats). These studies should allow improved risk assessments for genetic counseling of women with premutation or intermediate-size alleles. PMID- 12529856 TI - Protein phosphatase 2C is involved in the cAMP-dependent ciliary control in Paramecium caudatum. AB - Forward swimming of the Triton-extracted model of Paramecium is stimulated by cAMP. Backward swimming of the model induced by Ca(2+) is depressed by cAMP. Cyclic AMP and Ca(2+) act antagonistically in setting the direction of the ciliary beat. Some ciliary axonemal proteins from Paramecium caudatum are phosphorylated in a cAMP-dependent manner. In the presence of cAMP, axonemal 29- and 65-kDa polypeptides were phosphorylated by endogenous A-kinase in vitro. These phosphoproteins, however, were not dephosphorylated after in vitro phosphorylation, presumably because of the low endogenous phosphoprotein phosphatase activity associated with isolated axonemes. We purified the protein phosphatase that specifically dephosphorylated the 29- and 65-kDa phosphoproteins from Paramecium caudatum. The molecular weight of the protein phosphatase was 33 kDa. The protein phosphatase had common characteristics as protein phosphatase 2C (PP2C). The characteristics of the protein phosphatase were the same as those of the PP2C from Paramecium tetraurelia (PtPP2C) [Grothe et al., 1998: J. Biol. Chem. 273:19167-19172]. We concluded that the phosphoprotein phosphatase is the PP2C from Paramecium caudatum (PcPP2C). The PcPP2C markedly accelerated the backward swimming of the Triton-extracted model in the presence of Ca(2+). On the other hand, the PcPP2C slightly depressed the forward swimming speed. This indicates that the PP2C plays a role in the cAMP-dependent regulation of ciliary movement in Paramecium caudatum through dephosphorylation of 29- and/or 65-kDa regulatory phosphoproteins by terminating the action of cAMP. PMID- 12529855 TI - FOXL2 and BPES: mutational hotspots, phenotypic variability, and revision of the genotype-phenotype correlation. AB - Blepharophimosis syndrome (BPES), an autosomal dominant syndrome in which an eyelid malformation is associated (type I) or not (type II) with premature ovarian failure (POF), has recently been ascribed to mutations in FOXL2, a putative forkhead transcription factor gene. We previously reported 22 FOXL2 mutations and suggested a preliminary genotype-phenotype correlation. Here, we describe 21 new FOXL2 mutations (16 novel ones) through sequencing of open reading frame, 5' untranslated region, putative core promoter, and fluorescence in situ hybridization analysis. Our study shows the existence of two mutational hotspots: 30% of FOXL2 mutations lead to polyalanine (poly-Ala) expansions, and 13% are a novel out-of-frame duplication. In addition, this is the first study to demonstrate intra- and interfamilial phenotypic variability (both BPES types caused by the same mutation). Furthermore, the present study allows a revision of the current genotype-phenotype correlation, since we found exceptions to it. We assume that for predicted proteins with a truncation before the poly-Ala tract, the risk for development of POF is high. For mutations leading to a truncated or extended protein containing an intact forkhead and poly-Ala tract, no predictions are possible, since some of these mutations lead to both types of BPES, even within the same family. Poly-Ala expansions may lead to BPES type II. For missense mutations, no correlations can be made yet. Microdeletions are associated with mental retardation. We conclude that molecular testing may be carefully used as a predictor for POF risk in a limited number of mutations. PMID- 12529858 TI - Hemidesmosome protein dynamics in live epithelial cells. AB - Hemidesmosomes mediate stable anchorage of epithelial cells to laminin-5 in the basement membrane zone and have been likened to spot-welds. Indeed, it has been assumed that hemidesmosomes are not dynamic, at least when compared to other matrix adhesion sites including focal contacts. We tested this notion by monitoring the fate of green fluorescent protein (GFP)-tagged human integrin beta4 subunit (GFP-hbeta4) and GFP-tagged 180-kD human bullous pemphigoid (BP) autoantigen (GFP-BP180) in live cultures of 804G cells that assemble numerous mature hemidesmosomes. In subconfluent 804G cells, both GFP-hbeta4 and GFP-BP180 protein clusters are not stable but assemble into and disassemble out of cat paw like arrays at a relatively rapid rate. In confluent populations of 804G cells, although some cat paw-like clusters of both GFP-hbeta4 and GFP-BP180 are stable over periods of >60 min, other GFP-hbeta4 and GFP-BP180 protein arrays form and/or disappear during the same time period. Moreover, individual labeled particles show considerable motility in the plane of the membrane. Fluorescence recovery after photobleaching analyses provide a further indication of the dynamics of hemidesmosome proteins. In particular, bleached GFP-hbeta4 protein clusters in confluent cells recover signal within about 30 min, indicating that there is a relatively rapid turnover of hemidesmosome components in protein arrays clustered along the substratum attached surface of a cell. The rate of recovery is dependent on an intact microfilament system. In sharp contrast, bleached GFP-BP180 protein clusters in confluent cells fail to recover signal even when observed for longer than 60 min. To evaluate hemidesmosome protein dynamics in motile cells, we monitored GFP-hbeta4 and GFP-BP180 in 804G cells populating scrape wound sites in vitro. In these migratory cells, which lack mature hemidesmosomes, integrin beta4 subunit and BP180 protein clusters progressively assemble and disassemble into linear and cat-paw arrays. In summary, hemidesmosome protein clusters, like their counterparts in focal contacts, are dynamic. We discuss these results in relation to hemidesmosome functions. PMID- 12529857 TI - Acute effects of desmin mutations on cytoskeletal and cellular integrity in cardiac myocytes. AB - Mutations in desmin have been associated with a subset of human myopathies. Symptoms typically appear in the second to third decades of life, but in the most severe cases can manifest themselves earlier. How desmin mutations lead to aberrant muscle function, however, remains poorly defined. We created a series of four mutations in rat desmin and tested their in vitro filament assembly properties. RDM-G, a chimera between desmin and green fluorescent protein, formed protofilament-like structures in vitro. RDM-1 and RDM-2 blocked in vitro assembly at the unit-length filament stage, while RDM-3 had more subtle effects on assembly. When expressed in cultured rat neonatal cardiac myocytes via adenovirus infection, these mutant proteins disrupted the endogenous desmin filament to an extent that correlated with their defects in in vitro assembly properties. Disruption of the desmin network by RDM-1 was also associated with disruption of plectin, myosin, and alpha-actinin organization in a significant percentage of infected cells. In contrast, expression of RDM-2, which is similar to previously characterized human mutant desmins, took longer to disrupt desmin and plectin organization and had no significant effect on myosin or alpha-actinin organization over the 5-day time course of our studies. RDM-3 had the mildest effect on in vitro assembly and no discernable effect on either desmin, plectin, myosin, or alpha-actinin organization in vivo. These results indicate that mutations in desmin have both direct and indirect effects on the cytoarchitecture of cardiac myocytes. PMID- 12529859 TI - Linked regulation of motility and integrin function in activated migrating neutrophils revealed by interference in remodelling of the cytoskeleton. AB - Neutrophils migrate rapidly by co-ordinating regulation of their beta2-integrin adhesion with turnover of filamentous F-actin. The seven-protein Arp2/3 complex regulates actin polymerisation upon activation by proteins of the WASP-family. To investigate links between actin polymerisation, adhesion, and migration, we used a novel osmotic-shock method to load neutrophils with peptides: (1). WASP-WA and Scar-WA (which incorporate the actin- and Arp2/3-binding regions of WASP and Scar1), to compete with endogenous WASP-family members; (2). proline rich motifs (PRM) from the ActA protein of L. monocytogenes or from vinculin, which bind vasodilator-stimulated phosphoprotein (VASP), a regulator of cytoskeleton assembly. In a flow system, rolling-adherent neutrophils were stimulated with formyl tri-peptide. This caused rapid immobilisation, followed by migration with increasing velocity, supported by activated beta2-integrin CD11b/CD18. Loading ActA PRM (but not vinculin PRM) caused concentration-dependent reduction in migration velocity. At the highest concentration, unstimulated neutrophils had elevated F-actin and were rigid, but could not change their F-actin content or shape upon stimulation. Scar-WA also caused marked reduction in migration rate, but WASP-WA had a lesser effect. Scar-WA did not modify activation-dependent formation of F-actin or change in shape. However, a reduction in rate of downregulation of integrin adhesion appeared to contribute to impaired migration. These studies show that interference in cytoskeletal reorganisation that follows activation in neutrophils, can impair regulation of integrin function as well as motility. They also suggest a role of the Arp2/3 complex and WASP-family in co ordinating actin polymerisation and integrin function in migrating neutrophils. PMID- 12529860 TI - Centrosome and spindle pole body dynamics. Review and abstracts of the EMBO/EMBL Conference on Centrosomes and Spindle Pole Bodies, Heidelberg, September 13-17, 2002. AB - Five years after the first meeting held on Centrosomes and Spindle Pole Bodies, a second meeting was organized by Tano Gonzalez, Eric Karsenti, Kip Sluder, and Mark Winey in Heidelberg, Germany. Sponsored by the gracious European community (EMBO/EMBL), the meeting was both spectacular and exhausting. The wealth of information delivered, the plethora of model systems and unique approaches described, and the free exchange of information by a cooperative and excited community of scientists overwhelmed all participants. Even the best prepared scholars could not have anticipated the avalanche of data and insights that poured from the presentations from beginning to end. Daily posters by young and senior scientists added dimension to round out the well-planned series of presentations. The meeting began with opening remarks by Eric Karsenti and Michel Bornens who reminded participants of the historical questions of the field. Where does the centrosome come from? What are the mechanisms that control centrosome assembly and duplication? How is duplication coordinated with the cell cycle? Why do some cells have centrosomes, while others do not? What are the components of the centrosome? Does the centrosome play an important role in disease? PMID- 12529861 TI - Blocking of malaria parasite development in mosquito and fecundity reduction by midgut antibodies in Anopheles stephensi (Diptera: Culicidae). AB - Rabbits were immunized three times with extracts of Anopheles stephensi midgut. Immunized rabbits showed a high titer of antibodies when characterized by ELISA. We investigated the effect of anti-mosquito midgut antibodies on mosquito fecundity, longevity, mortality, engorgement, and the development of the malaria parasite in mosquitoes. Fecundity was reduced significantly (38%) and similarly hatchability by about 43.5%. There was no statistically significant effect on mortality, longevity, and engorgement. When the mosquito blood meal contained anti-midgut antibodies, fewer oocysts of Plasmodium vivax developed in the mosquito midgut and the proportion of mosquitoes becoming infected was significantly reduced. We also found that the midgut antibodies inhibit the development and/or translocation of the sporozoites. Antisera raised against midgut of A. stephensi recognized eight polypeptides (110, 92, 70, 45, 38, 29, 15, 13 kDa) by Western blotting. Cross-reactive antigens/epitopes present in other tissues of A. stephensi were also examined both by Western blotting and in vivo ELISA. Together, these observations open an avenue for research toward the development of a vector-based malaria parasite transmission blocking vaccine and/or anti-mosquito vaccine. PMID- 12529862 TI - The bacterium Xenorhabdus nematophilus depresses nodulation reactions to infection by inhibiting eicosanoid biosynthesis in tobacco hornworms, Manduca sexta. AB - The bacterium, Xenorhabdus nematophilus, is a virulent insect pathogen. We tested the hypothesis that this bacterium impairs insect cellular immune defense reactions by inhibiting biosynthesis of eicosanoids involved in mediating cellular defense reactions. Fifth instar tobacco hornworms, Manduca sexta, produced melanized nodules in reaction to challenge with living and heat-killed X. nematophilus. However, the nodulation reactions were much attenuated in insects challenged with living bacteria (approximately 20 nodules/larva for living bacteria vs. approximately 80 nodules/larva in insects challenged with heat-killed bacteria). The nodule-inhibiting action of living X. nematophilus was due to a factor that was present in the organic, but not aqueous, fraction of the bacterial cultural medium. The nodule-inhibiting factor in the organic fraction was labile to heat treatments. The immunodepressive influence of the factor in the organic fraction was reversed by treating challenged hornworms with arachidonic acid. The factor also depressed nodulation reactions to challenge with the plant pathogenic bacteria, Pseudomonas putida and Ralstonia solanacearum. These findings indicate that one or more factors from X. nematophilus depress nodulation reactions in tobacco hornworms by inhibiting eicosanoid biosynthesis. PMID- 12529863 TI - Nutritional plasticity of the predatory ladybeetle Harmonia axyridis (Coleoptera: Coccinellidae): comparison between natural and substitution prey. AB - The predatory coccinellid Harmonia axyridis is a polyphagous species, efficient at controlling certain aphid species and already commercialized in Europe for that purpose. The complete development of this predator can be accomplished using the aphid Acyrthosiphon pisum or Ephestia kuehniella eggs as substitution prey. Biochemical analyses were conducted on the proteins, lipids, and carbohydrates of these two different prey species. E. kuehniella eggs were 2 times richer in amino acids than A. pisum adults (12% of the fresh weight vs. 6%). E. kuehniella eggs were 3 times richer in lipids than the aphids but, on the contrary, the aphids were 1.5 times richer in glycogen. The impact of these two kinds of food on the body composition of the coccinellid was evaluated to appreciate the degree of nutritional plasticity of the coccinellid. The composition of the coccinellids feeding either on E. kuehniella eggs or on aphids was compared for amino acid, fatty acid and glycogen contents, revealing a good capability of H. axyridis to develop on foods that are very different in their biochemical composition. Nevertheless, when fed on aphids, the crude protein content of the predator was reduced and the lipid content decreased by a factor of two, with a change in amino and fatty acid patterns. Some biological parameters, such as larval mortality, adult weight, and fecundity, were modified according to the food eaten. PMID- 12529864 TI - Purification and cDNA cloning of inducible antibacterial peptides from Protaetia brevitarsis (Coleoptera). AB - Three antibacterial peptides, named protaetins 1, 2, and 3, were purified and characterized from immunized larval hemolymph of Protaetia brevitarsis, a fruit tree pest in Korea. Also, protaetin 1 was cloned. Acid extraction, gel filtration, preparative acid-urea PAGE, and reversed-phase FPLC were used for purification of peptides. Protaetins 1 and 3 had molecular masses of 7.5 and 12 kDa on Tricine SDS-PAGE, respectively, and the molecular mass of protaetin 2 was 9,283.95 Da as determined by MALDI-TOF mass spectrometry. In an antibacterial assay, protaetins showed antibacterial activities against a panel of Gram positive and -negative bacteria. For the RT-PCR (reverse transcription polymerase chain reaction) to obtain the complete primary sequence, the primer was designed according to the N-terminal amino acid sequence of protaetin 1. Amino acid sequence homology of protaetin 1 with holotricin 2, an antibacterial peptide from Holotrichia diomphalia, showed 99% identity. Northern blot analysis showed that the protaetin 1 gene was strongly expressed in the fat body after Escherichia coli injection, but not in normal fat body. Also, it was expressed in the gut, but was much weaker after immunization. PMID- 12529865 TI - Polydnavirus of Campoletis chlorideae: characterization and temporal effect on host Helicoverpa armigera cellular immune response. AB - Polydnavirus was isolated from oviduct calyx in the parasitoid wasp Campoletis chlorideae (Hymenoptera: Ichneumonidae), and termed CcIV. The virus particles consist of fusiform nucleocapsids surrounded by two unit membrane envelopes. The DNAs purified from these viruses were found diversified in molecular weight and existed in nonequimolar concentrations. At least 20 different-sized bands were present after electrophoresis, and they ranged from approximately 3 to 26 kb. Persistence and gene expression of CcIV were examined in parasitized and virus injected Helicoverpa armigera larvae. Viral DNA could be detected in the hemocyte of H. armigera at 30 min post-parasitization (p.p.), and persisted for 6 days. While no viral DNA increase was found, CcIV transcripts were first detected in host hemocytes at day 1 p.p. and continued for 5 days. Similar transcripts were observed in hemocytes from larvae that had been injected with calyx fluid or CcIV 24 h earlier. CcIV viral DNAs hybridized only with certain first-strand cDNAs from hemocytes, suggesting that only part of the CcIV genome was expressed in H. armigera. The pattern of CcIV gene expression was consistent with that of the inhibition of encapsulation for Sephadex G-10 and parasitoid eggs by host larvae. The recovery of host immune response at day 4 p.p. indicated that CcIV exhibited a partial and temporal effect on the host immune system and the developing parasitoid appeared to avoid encapsulation via different mechanisms. PMID- 12529867 TI - A parametric model for long-term follow-up data from phase III breast cancer clinical trials. AB - We propose a parametric version of a univariate gamma frailty model. The proposed model is shown to be flexible enough to model long-term follow-up survival data from breast cancer clinical trials when the treatment effect diminishes as time progresses, a case for which neither the proportional hazards nor proportional odds assumptions are satisfied. The observed information matrix is computed to evaluate the variances of parameter estimates. A simple parametric test statistic to test proportional odds assumption is also constructed. The model is applied to a data set from a phase III clinical trial on breast cancer. PMID- 12529868 TI - Incremental net benefit in randomized clinical trials with quality-adjusted survival. AB - Owing to induced dependent censoring, estimating mean costs and quality-adjusted survival in a cost-effectiveness comparison of two groups using standard life table methods leads to biased results. In this paper we propose methods for estimating the difference in mean costs and the difference in mean effectiveness, together with their respective variances and covariance in the presence of dependent censoring. We consider the situation in which the measure of effectiveness is either the probability of surviving a duration of interest or mean quality-adjusted survival time over a duration of interest. The methods are illustrated in an example using an incremental net benefit analysis. PMID- 12529869 TI - Non-linear models for the relation between cardiovascular risk factors and intake of wine, beer and spirits. AB - It is generally accepted that moderate consumption of alcohol is associated with a reduced risk of coronary heart disease (CHD). It is not clear however whether this benefit is derived through the consumption of a specific beverage type, for example, wine. In this paper the associations between known CHD risk factors and different beverage types are investigated using a novel approach with non-linear modelling. Two types of model are proposed which are designed to detect differential effects of beverage type. These may be viewed as extensions of Box and Tidwell's power-linear model. The risk factors high density lipoprotein cholesterol, fibrinogen and systolic blood pressure are considered using data from a large longitudinal study of British civil servants (Whitehall II). The results for males suggest that gram for gram of alcohol, the effect of wine differs from that of beer and spirits, particularly for systolic blood pressure. In particular increasing wine consumption is associated with slightly more favourable levels of all three risk factors studied. For females there is evidence of a differential relationship only for systolic blood pressure. These findings are tentative but suggest that further research is required to clarify the similarities and differences between the results for males and females and to establish whether either of the models is the more appropriate. However, having clarified these issues, the apparent benefit of consuming wine instead of other alcoholic beverages may be relatively small. PMID- 12529870 TI - Viral load detectability profiles for HIV infection. AB - The introduction of potent antiretroviral therapies for treatment of HIV infection typically results in a dramatic reduction in plasma HIV RNA concentration, often to levels undetectable by current measurement practices. However, although a high proportion of patients achieve 'undetectability', many then experience a return to a state of detectability at a later date. As evaluation of virologic response provides a useful measure of therapy efficacy, it is of interest to estimate the proportions of cases with undetectable viral load over time following commencement of treatment. These proportions depend on the rates of transition from detectability to undetectability and subsequent return to detectability, and may be related to covariates or risk factors, possibly differing in both transitions. We consider construction of detectability profiles as estimates of these proportions, based on parametric modelling of the component survival distributions. The method is applied to an examination of the effects of baseline CD4 T-cell lymphocyte counts on virologic response to therapy amongst patients of the Western Australian HIV Cohort Study. PMID- 12529871 TI - Marginal versus joint Box-Cox transformation with applications to percentile curve construction for IgG subclasses and blood pressures. AB - When age-specific percentile curves are constructed for several correlated variables, the marginal method of handling one variable at a time has typically been used. We address the question, frequently asked by practitioners, of whether we can achieve efficiency gains by joint estimation. We focus on a simple but common method of Box-Cox transformation and assess the statistical impact of a joint transformation to multivariate normality on the percentile curve estimation for correlated variables. We find that there is little gain from the joint transformation for estimating percentiles around the median but a noticeable reduction in variances is possible for estimating extreme percentiles that are usually of main interest in medical and biological applications. Our study is motivated by problems in constructing percentile charts for IgG subclasses of children and for blood pressures in adult populations, both of which are discussed in the paper as examples, and yet our general findings are applicable to a wide range of other problems. PMID- 12529872 TI - Inference for a linear regression model with an interval-censored covariate. AB - Interval-censored observations of a response variable are a common occurrence in medical studies, and usually result when the response is the elapsed time until some event whose occurrence is periodically monitored. In this paper we consider a multivariate regression setting in which the explanatory variable is interval censored. Use of an ad hoc method of analysis for such data, such as taking the midpoint of the interval-censored covariate and applying ordinary least-squares, is not in general valid. We develop a likelihood approach, together with a two step conditional algorithm, to jointly estimate the regression coefficients as well as the marginal distribution of the covariate. The resulting estimators are asymptotically normal. The performance of the method is assessed via simulations, and illustrated using data from a recent HIV/AIDS clinical trial to assess the association between waiting time between indinavir failure and subsequent viral load at enrolment. Extensions of the procedure to other parametric distributions are discussed. PMID- 12529873 TI - Transitions in an imperfectly observed binary variable: depressive symptomatology in adolescents. AB - We describe a methodology for analysing transitions over time in a binary outcome variable that is subject to misclassification (that is, measurement error). Logistic regression models for transition events in the true underlying state are combined with estimates of probabilities of misclassification of the underlying state. The model is based on the Markovian assumption that the probabilities of transition in the underlying state at a given time depend only on the underlying state at the previous time. Hence we estimate odds-ratio effects for transitions that are adjusted for the effect of misclassification. Comparing these adjusted estimates with estimates that are obtained without taking misclassification into account indicates that the latter can be biased either toward or away from the null. For the estimates to exist, certain restrictions on the observed data and misclassification probabilities need to be met. If these restrictions are not satisfied then the conclusion from the analysis is that all observed transition events can be explained solely by the error in outcome assessment, in which case it is likely that an aspect of the model is incorrect. The motivation for this work comes from an analysis of transitions in depression status for a cohort of Australian teenagers participating in a longitudinal study of adolescent health. PMID- 12529874 TI - Applications of continuous time hidden Markov models to the study of misclassified disease outcomes. AB - Disease progression in prospective clinical and epidemiological studies is often conceptualized in terms of transitions between disease states. Analysis of data from such studies can be complicated by a number of factors, including the presence of individuals in various prevalent disease states and with unknown prior disease history, interval censored observations of state transitions and misclassified measurements of disease states. We present an approach where the disease states are modelled as the hidden states of a continuous time hidden Markov model using the imperfect measurements of the disease state as observations. Covariate effects on transitions between disease states are incorporated using a generalized regression framework. Parameter estimation and inference are based on maximum likelihood methods and rely on an EM algorithm. In addition, techniques for model assessment are proposed. Applications to two binary disease outcomes are presented: the oral lesion hairy leukoplakia in a cohort of HIV infected men and cervical human papillomavirus (HPV) infection in a cohort of young women. Estimated transition rates and misclassification probabilities for the hairy leukoplakia data agree well with clinical observations on the persistence and diagnosis of this lesion, lending credibility to the interpretation of hidden states as representing the actual disease states. By contrast, interpretation of the results for the HPV data are more problematic, illustrating that successful application of the hidden Markov model may be highly dependent on the degree to which the assumptions of the model are satisfied. PMID- 12529875 TI - Estimating cumulative probabilities from incomplete longitudinal binary responses with application to HIV vaccine trials. AB - When describing longitudinal binary response data, it may be desirable to estimate the cumulative probability of at least one positive response by some time point. For example, in phase I and II human immunodeficiency virus (HIV) vaccine trials, investigators are often interested in the probability of at least one vaccine-induced CD8+ cytotoxic T-lymphocyte (CTL) response to HIV proteins at different times over the course of the trial. In this setting, traditional estimates of the cumulative probabilities have been based on observed proportions. We show that if the missing data mechanism is ignorable, the traditional estimator of the cumulative success probabilities is biased and tends to underestimate a candidate vaccine's ability to induce CTL responses. As an alternative, we propose applying standard optimization techniques to obtain maximum likelihood estimates of the response profiles and, in turn, the cumulative probabilities of interest. Comparisons of the empirical and maximum likelihood estimates are investigated using data from simulations and HIV vaccine trials. We conclude that maximum likelihood offers a more accurate method of estimation, which is especially important in the HIV vaccine setting as cumulative CTL responses will likely be used as a key criterion for large scale efficacy trial qualification. PMID- 12529876 TI - A statistical perspective on gene expression data analysis. AB - Rapid advances in biotechnology have resulted in an increasing interest in the use of oligonucleotide and spotted cDNA gene expression microarrays for medical research. These arrays are being widely used to understand the underlying genetic structure of various diseases, with the ultimate goal to provide better diagnosis, prevention and cure. This technology allows for measurement of expression levels from several thousands of genes simultaneously, thus resulting in an enormous amount of data. The role of the statistician is critical to the successful design of gene expression studies, and the analysis and interpretation of the resulting voluminous data. This paper discusses hypotheses common to gene expression studies, and describes some of the statistical methods suitable for addressing these hypotheses. S-plus and SAS codes to perform the statistical methods are provided. Gene expression data from an unpublished oncologic study is used to illustrate these methods. PMID- 12529877 TI - Stability of protein production from recombinant mammalian cells. AB - One of the most important criteria for successful generation of a therapeutic protein from a recombinant cell is to obtain a cell line that maintains stability of production. If this is not achieved it can generate problems for process yields, effective use of time and money, and for regulatory approval of products. However, selection of a cell line that sustains stability of production over the required time period may be difficult to achieve during development of a therapeutic protein. There are several studies in the literature that have reported on the instability of protein production from recombinant cell lines. The causes of instability of production are varied and, in many cases, the exact molecular mechanisms are unknown. The production of proteins by cells is modulated by molecular events at levels ranging from transcription, posttranscriptional processing, translation, posttranslational processing, to secretion. There is potential for regulation of stability of protein production at many or all of these stages. In this study we review published information on stability of protein production for three industrially important cell lines: hybridoma, Chinese hamster ovary (CHO), and nonsecreting (NS0) myeloma cell lines. We highlight the most likely molecular loci at which instability may be engendered and indicate other areas of protein production that may affect stability from mammalian cells. We also outline approaches that could help to overcome the problems associated with unpredictable expression levels and maximized production, and indicate the consequences these might have for stability of production. PMID- 12529878 TI - Modeling the performance of pilot-scale countercurrent chromatography: scale-up predictions and experimental verification of erythromycin separation. AB - Biosynthesis of polyketide antibiotics, such as erythromycin A (EA), can result in the formation of analogues of the main compound that are chemically and structurally extremely similar. The large-scale purification of these antibiotics by conventional high-performance liquid chromatography (HPLC) can be prohibitively expensive due to the large volume of both solvent and adsorbent required. This study examines the feasibility of using a novel pilot-scale countercurrent chromatography (CCC) machine as an alternative to HPLC. CCC is a low-pressure (typically <4000 kN m(-2)) liquid-liquid chromatographic technique that allows the separation of solutes on the basis of their partitioning between two immiscible liquid phases. The effects of mobile phase flow rate, column rotational speed, and sample injection volume on the attainable yield and purity of EA were investigated. Our results show that, at a mobile phase flow rate of 40 mL min(-1), a rotational speed of 1200 rpm, and an injection volume of 100 mL (10 g total erythromycin), EA could be satisfactorily fractionated with a purity of approximately 92% (w/w) and a recovery yield of approximately 100% (w/w). The total solute throughput was estimated to be 0.41 kg day(-1). More importantly, we demonstrated simple and predictive linear scale-up of the CCC separation based on data obtained from a single laboratory-scale CCC chromatogram, and verified this experimentally. The retention time and peak width of the target compound at the pilot scale could be predicted to within 4% for operation at a range of mobile phase flow rates and injection volumes. This predictable nature of CCC separations, unlike HPLC methods, can greatly reduce process development times and enable a complete process-scale operating scenario to be planned. PMID- 12529880 TI - High-cell-density fed-batch cultivation of the docosahexaenoic acid producing marine alga Crypthecodinium cohnii. AB - The heterotrophic marine alga Crypthecodinium cohnii is known to produce docosahexaenoic acid (DHA), a polyunsaturated fatty acid with food and pharmaceutical applications, during batch cultivation on complex media containing sea salt, yeast extract, and glucose. In the present study, fed-batch cultivation was studied as an alternative fermentation strategy for DHA production. Glucose and acetic acid were compared as carbon sources. For both substrates, the feed rate was adapted to the maximum specific consumption rate of C. cohnii. In glucose-grown cultures, this was done by maintaining a significant glucose concentration (between 5 and 20 g/L) throughout fermentation. In acetic acid grown cultures, the medium feed was automatically controlled via the culture pH. A feed consisting of acetic acid (50% w/w) resulted in a higher overall volumetric productivity of DHA (r(DHA)) than a feed consisting of 50% (w/v) glucose (38 and 14 mg/L/h, respectively). The r(DHA) was further increased to 48 mg/L/h using a feed consisting of pure acetic acid. The latter fermentation strategy resulted in final concentrations of 109 g/L dry biomass, 61 g/L lipid, and 19 g/L DHA. These are the highest biomass, lipid, and DHA concentrations reported to date for a heterotrophic alga. Vigorous mixing was required to sustain aerobic conditions during high-cell-density cultivation. This was complicated by culture viscosity, which resulted from the production of viscous extracellular polysaccharides. These may present a problem for large-scale industrial production of DHA. Addition of a commercial polysaccharide-hydrolase preparation could decrease the viscosity of the culture and the required stirring. PMID- 12529879 TI - Cell affinity separations using magnetically stabilized fluidized beds: erythrocyte subpopulation fractionation utilizing a lectin-magnetite support. AB - A magnetically stabilized fluidized bed is used to separate erythrocyte subpopulations. Binding specificity was obtained by immobilizing the lectin Helix pomatia Agglutinin (HpA) or Griffonia simplicifolia I (GSI) onto a magnetite containing support. Separation of type A and type O erythrocytes with the lectin HpA was particularly effective, leading to a 94% purity of retained type A erythrocytes. A 3.1 +/- 0.6 log removal of type A erythrocytes was also accomplished leading to a 99.7% +/- 0.4% purity and 95% +/- 7% yield of type O erythrocytes in the collected effluent. Elution of the purified cells was accomplished using fluidization in the presence of a sugar competing for the lectin-erythrocyte binding site. A mathematical model based on the depth filtration model of Putnam and Burns (Chem Eng Sci 1997;52(1):93-105) was extended to include multicomponent cell adhesion. This filtration model is the first to take into account the finite binding capacity of the chromatographic support and is used to characterize the cell binding behavior and to determine optimal parameters and conditions that lead to high capacities and selectivities. Model parameter values and observations from in situ adsorption studies suggest that the non-spherical shape of the magnetite-based support allows for a more efficient utilization of the support surface area than the spherical shape. Using a 1.5-cm diameter laboratory column and realistic parameter values, the processing rates of the system are predicted to be at least an order of magnitude greater than the 10(8)/h cells that can typically be processed in packed bed cell affinity chromatography (CAC) systems. PMID- 12529881 TI - Parameter scanning ultrafiltration: rapid optimisation of protein separation. AB - High-resolution fractionation of proteins using ultrafiltration is feasible only at highly optimised conditions. Conventional process optimisation methodology demands both time and material. Pulsed sample injection ultrafiltration has been suggested as a rapid process optimisation technique. In the present work the scope of this technique is further extended by "parameter scanning ultrafiltration," which involves continuous change of a process parameter (e.g., pH, salt concentration). The time and material consumption are thus further reduced. The technique was validated using different proteins and membranes. Sieving coefficients at different pH and salt concentration were compared to those obtained in fixed parameter ultrafiltration experiments. As fractionation case studies the separation of monoclonal antibody from bovine serum albumin and separation of human IgG from human serum albumin were examined. PMID- 12529882 TI - Use of the two-liquid phase concept to exploit kinetically controlled multistep biocatalysis. AB - The two-liquid phase concept was used to develop a whole cell biocatalytic system for the efficient multistep oxidation of pseudocumene to 3,4 dimethylbenzaldehyde. Recombinant Escherichia coli cells were employed to express the Pseudomonas putida genes encoding xylene monooxygenase, which catalyzes the multistep oxygenation of one methyl group of toluene and xylenes to corresponding alcohols, aldehydes, and acids. A fed-batch based two-liquid phase bioconversion was established with bis(2-ethylhexyl)- phthalate as organic carrier solvent and a phase ratio of 0.5; the product formation pattern, the impact of the nutrient feeding strategy, and the partitioning behavior of the reactants were studied. On the basis of the favorable conditions provided by the two-liquid phase system, engineering of the initial pseudocumene concentration allowed exploiting the complex kinetics of the multistep reaction for the exclusive production of 3,4 dimethyl- benzaldehyde. Further oxidation of the product to 3,4-dimethylbenzoic acid could be inhibited by suitable concentrations of pseudocumene or 3,4 dimethylbenzyl alcohol. The optimized biotransformation setup includes a completely defined medium with high iron content and a nutrient feeding strategy that avoids severe glucose limitation as well as high inhibitory glucose levels. Using such a system on a 2-liter scale, we were able to produce, within 14.5 h, 30 g of 3,4-dimethylbenzaldehyde as predominant reactant in the organic phase and reached a maximal productivity of 1.6 g per liter liquid volume per hour. The present study implicates that the two-liquid phase concept is an efficient tool to exploit the kinetics of multistep biotransformations in general. PMID- 12529883 TI - Preparation and characterization of reactive and stable glucose oxidase containing liposomes modulated with detergent. AB - Glucose oxidase-containing liposomes (GOL) as well as detergent-modulated glucose oxidase-containing liposomes were prepared and characterized, focusing not only on the reactivity of the liposomes upon external addition of glucose but also on the leakage of the entrapped glucose oxidase (GO) from the liposomes with the aim of developing a reactive and stable liposomal GO system. The membranes of the GOL prepared were composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and modulated with either Triton X-100 or cholate. In the absence of added detergent, no GO leakage from the GOL was observed while its enzymatic activity was very low (low glucose permeability). As detergent-modulated liposomes, mixed POPC/Triton X-100 and mixed POPC/cholate liposomes (abbreviated as TL and CL, respectively) were prepared at different effective detergent/POPC molar ratios (R(e)) ranging from R(e) = 0 to R(e) = R(e) (sat) (R(e) (sat) is the critical value of R(e) at which the liposome membrane is saturated with detergent). The reactivity of GO-loaded TL (abbreviated as GOTL) or GO-loaded CL (GOCL) increased drastically with increase in the respective detergent content in the liposomes. In the case of GOTL, at R(e) (sat) = 0.40, a high reactivity was measured with a simultaneous high extent of GO leakage, suggesting that the observed enzymatic reaction was catalyzed mainly by leaked GO, caused by the interaction of Triton X 100 with the POPC membrane. On the other hand, GOCL prepared at R(e) (sat) = 0.43 showed relatively high reactivity with only a small extent of GO leakage, suggesting that most of the enzyme reaction was limited by the glucose permeation across the bilayers of GOCL. The GO leakage from GOCL was found to occur mostly during the rearrangement of the liposomal membrane during the preparation of the GOCL (mixing the GOL and cholate). Fluorescence polarization measurements of membrane-associated DPH (1,6-diphenyl-1,3,5-hexatriene) indicated that CL prepared by modifying POPC with cholate did not lead to a drastic change in membrane fluidity, indicating that the interacting cholate molecules did not penetrate deeply into the POPC bilayers. In summary, it was clearly shown that the membrane permeability of GOL can be quite simply modulated by mixing it with a certain amount of cholate to form highly reactive and stable GOCL with minimal enzyme leakage. PMID- 12529884 TI - Immobilization of D-ribulose-1,5-bisphosphate carboxylase/oxygenase: a step toward carbon dioxide fixation bioprocess. AB - Immobilization of D-ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from spinach leaves is described. This enzyme enables the fixation of carbon dioxide on a five-carbon sugar D-ribulose-1,5-bisphosphate (RuBP). Two different immobilization methods were employed: dicyclohexylcarbodiimide coupling on nylon membrane matrix and dimethylpimelimidate immobilization on protein A agarose. The reusability of immobilized enzymes, coupling efficiency, and temperature-activity relationship of soluble and immobilized Rubisco are presented. The immobilization imparted greater thermal and storage stability. The thermal deactivation rates of the immobilized enzymes were considerably lower than those of the soluble enzyme. PMID- 12529885 TI - Application of a gratuitous induction system in Kluyveromyces lactis for the expression of intracellular and secreted proteins during fed-batch culture. AB - A gratuitous induction system in the yeast Kluyveromyces lactis was evaluated for the expression of intracellular and extracellular products during fed-batch culture. The Escherichia coli lacZ gene (beta-galactosidase; intracellular) and MFalpha1 leader-BPTI cassette (bovine pancreatic trypsin inhibitor; extracellular) were placed under the control of the inducible K. lactis LAC4 promotor, inserted into partial-pKD1 plasmids, and transformed into a ga1-209 K. lactis strain. To obtain a high level of production, culture conditions for growth and expression were initially evaluated in tube cultures. A selective medium containing 5 g/L glucose (as carbon source) and 0.5 g/L galactose (as inducer) demonstrated the maximum activity of both beta-galactosidase and secreted BPTI. This level of expression had no significant effect on the growth of the recombinant cells; growth rate dropped by approximately 11%, whereas final biomass concentrations remained the same. In shake-flask culture, biomass concentration, beta-galactosidase activity, and BPTI secreted activity were 4 g/L, 7664 U/g dry cell, and 0.32 mg/L, respectively. Fed-batch culture (with a high glucose concentration and a low galactose [inducer] concentration feed) resulted in a 6.5-fold increase in biomass, a 23-fold increase in beta galactosidase activity, and a 3-fold increase in BPTI secreted activity. The results demonstrate the success of gratuitous induction during high-cell-density fed-batch culture of K. lactis. A very low concentration of galactose feed was sufficient for a high production level. PMID- 12529886 TI - Propagation of viruses on micropatterned host cells. AB - We have developed a technique to characterize the in vitro propagation of viruses. Microcontact printing was used to generate linear arrays of alkanethiols on gold surfaces, which served as substrates for the patterned culture of baby hamster kidney (BHK-21) cells. Vesicular stomatitis virus (VSV) was added to unpatterned cell reservoirs adjacent to the patterned cells and incubated, setting in motion a continuously advancing viral infection into the patterned cells. At different incubation times, multiple arrays were chemically fixed to stop the viral propagation. Viral propagation distances into the patterned cells were determined by indirect immunofluorescent labeling and visualization of the VSV surface glycoprotein (G). The infection spread at approximately 50 microm/h in the 140-microm lines. Moreover, different temporal stages of the infection process were simultaneously visualized along individual lines. These stages included initiation of infection, based on G protein expression; cell-cell fusion, based on virus-induced clustering of cell nuclei; and cytoskeletal degradation, based on localized release of cells from the surface. This work sets a foundation for parallel, high-throughput characterization of viral and cellular processes. PMID- 12529887 TI - Enzymatic hydrolysis of water-soluble wheat arabinoxylan. 1. Synergy between alpha-L-arabinofuranosidases, endo-1,4-beta-xylanases, and beta-xylosidase activities. AB - Hydrolysis of arabinoxylan is an important prerequisite for improved utilization of wheat hemicellulose in the ethanol fermentation industry. This study investigates the individual and combined efficiencies of three commercial, cellulytic and hemicellulytic enzyme preparations, Celluclast 1.5 L, Ultraflo L, and Viscozyme L, in catalyzing the liberation of arabinose and xylose from water soluble wheat arabinoxylan. Ultraflo L was the best enzyme preparation for releasing arabinose, liberating 53 wt% of the theoretical maximum after 48 h of reaction (10 wt% enzyme/substrate ratio, 40 degrees C, pH 6). Celluclast 1.5 L was superior to the other enzyme preparations in releasing xylose, liberating 26 wt% of the theoretical maximum after 48 h of reaction (10 wt% enzyme/substrate ratio, 50 degrees C, pH 5). The 50:50 mixtures of the enzyme preparations showed no synergistic cooperation in arabinose release, but a synergistic interaction in xylose release was found between Ultraflo L and Celluclast 1.5 L. On the basis of high-performance anion exchange chromatography (HPAEC) analysis of the hydrolysates after enzymatic reaction, we propose that the observed synergism between Celluclast 1.5 L and Ultraflo L is the result of positive interaction between alpha-L-arabinofuranosidase and endo-1,4-beta-xylanase activities present in Ultraflo L that released arabinose, xylobiose and xylotriose, and beta xylosidase activities in Celluclast 1.5 L, capable of catalyzing the hydrolysis of xylobiose and xylotriose to xylose. PMID- 12529888 TI - Transglutaminase-catalyzed synthesis of trypsin-cyclodextrin conjugates: kinetics and stability properties. AB - Bovine pancreatic trypsin was modified by the mono-6-amino-6-deoxy derivatives of alpha-, beta-, and gamma-cyclodextrin through a transglutaminase-catalyzed reaction. The trypsin-cyclodextrin conjugates, containing about 3 mol of oligosaccharide per mole of protein, were tested for their catalytic and stability properties. The specific esterolytic activity and the kinetics constants of trypsin were significantly improved following the transglutaminase induced structural modifications. Trypsin-cyclodextrin conjugates were also found markedly (sixfold) more resistant to autolytic degradation at alkaline pH, and their thermal stability profile was improved by about 16 degrees C. Moreover, they were particularly resistant to heat inactivation when treated at different temperatures ranging from 45 degrees C to 70 degrees C for different periods of time. PMID- 12529889 TI - Potential inhibitors from wet oxidation of wheat straw and their effect on ethanol production of Saccharomyces cerevisiae: wet oxidation and fermentation by yeast. AB - Alkaline wet oxidation (WO) (using water, 6.5 g/L sodium carbonate and 12 bar oxygen at 195 degrees C) was used as pretreatment method for wheat straw (60 g/L), resulting in a hydrolysate and a cellulosic solid fraction. The hydrolysate consisted of soluble hemicellulose (8 g/L), low-molecular-weight carboxylic acids (3.9 g/L), phenols (0.27 g/L = 1.7 mM) and 2-furoic acid (0.007 g/L). The wet oxidized wheat straw hydrolysate caused no inhibition of ethanol production by Saccharomyces cerevisiae ATCC 96581. Nine phenols and 2-furoic acid, identified to be present in the hydrolysate, were each tested in concentrations of 50-100 times the concentration found in the hydrolysate for their effect on fermentation by yeast. At these high concentrations (10 mM), 4-hydroxybenzaldehyde, vanillin, 4-hydroxyacetophenone and acetovanillone caused a 53-67% decrease in the volumetric ethanol productivity in S. cerevisiae compared to controls with an ethanol productivity of 3.8 g/L. The phenol acids (4-hydroxy, vanillic and syringic acid), 2-furoic acid, syringaldehyde and acetosyringone were less inhibitory, causing a 5-16% decrease in ethanol productivity. By adding the same aromatic compounds to hydrolysate (10 mM), it was shown that syringaldehyde and acetovanillone interacted negatively with hydrolysate components on the ethanol productivity. Fermentation in WO hydrolysate, that had been concentrated 6 times by freeze-drying, lasted 4 hours longer than in regular hydrolysate; however, the ethanol yield was the same. The longer fermentation time could not be explained by an inhibitory action of phenols alone, but was more likely caused by inhibitory interactions of phenols with carboxylic acids, such as acetic and formic acid. PMID- 12529890 TI - Intracellular Ca2+ imaging for micropatterned cardiac myocytes. AB - The patterning of cardiac myocytes on a micron scale ( approximately 5 microm) was achieved by microcontact printing of fibronectin onto a hydrophobically pretreated glass substrate. The patterned cardiac myocytes conjugated with each other by forming a gap junction, as judged from the synchronized Ca(2+) transition over the pattern, and thus simultaneously contracted. The dynamic change of the Ca(2+) concentration within the patterned tissue was analyzed quantitatively during successive contraction and relaxation using a Nipkow-type high-speed confocal microscope. PMID- 12529891 TI - Principles of preventive care. AB - Clinicians who wish to ensure that their prevention practices are current and appropriate should follow the following steps: Identify a source of prevention recommendations that is evidence-based and which has a perspective that matches that of your practice. Periodically review current recommendations to keep up with changing science. When a new screening test is introduced, ask whether it has been proven to improve important health outcomes or simply been shown to detect more disease. Consider what potential harms might be associated with screening. Acknowledge the role of the patient in decisions in areas of uncertainty. At the same time, educate patients that additional tests carry risks as well as benefits. Approach prevention as a package of important services. Educate patients of the need to address all important health issues rather than focusing on a single issue. Examine how well you are delivering high priority services. PMID- 12529892 TI - Cultural factors in preventive care: African-Americans. AB - In summary, the implications for healthcare practitioners in caring for African American patients in a culturally sensitive manner include: Gaining trust, and understanding the historical distrust of the health care system Understanding and employing the kinship web in decisions regarding screening and treatment Involving the church in developing and delivering prevention and care messages Asking patients about the meaning of words or phrases Asking patients about the use of alternative medicines and herbs Tailoring messages about prevention to depictions of real life situations Paying attention to body language and other nonverbal communication. PMID- 12529893 TI - Cultural factors in preventive care: Asian-Americans. AB - The Asian-American population in the United States continues to grow rapidly. In 1990, there were 6.6 million Asian Americans in this country, representing 2.7% of the population. The 2000 Census showed the Asian population has grown to 10.1 million, now 3.6% of the population. A breakdown of the major ethnic groups revealed: Chinese 24%, Filipino 18%, Asian-Indians 16%, Vietnamese 11%, Koreans 10%, and Japanese 8%. According to the 1990 Census, 66% of the Asian-American population was foreign-born, 40% reported that they do not speak English 'very well', and 25% were in linguistically isolated households. Due to the differences in their ethnic background, language barriers, cultural beliefs, socioeconomic status and distrust in Western medicine, providing care to this population constitutes a formidable challenge for primary care clinicians. PMID- 12529894 TI - Cultural factors in preventive care: Latinos. AB - For many, the term "Hispanic" places undue emphasis on the European influence of Spanish colonialism and may even have negative connotations for some. "Latino" is a more encompassing term that gives recognition to the influences of the indigenous and African cultures on modern day Latin Americans. Nevertheless, recognition of typical Latino attitudes and beliefs may assist health care providers. Poverty, unemployment, and low level of education usually account for adverse health in this population. Anti-immigrant sentiment and discrimination in health care and education add adversity to the immigrant's experience. Lack of health insurance and access to quality health care typically plague the adult immigrant. For many, the nearest emergency department is their only source of medical care. PMID- 12529895 TI - Preconception and prenatal care. AB - Advances in preconception and prenatal care have been successful in reducing risk in a number of areas. Folic acid supplementation, abstinence from alcohol, tight glycemic control in pre-gestational diabetics, and the administration of rhogham all have been successful in reducing individual risks. Unfortunately, overall perinatal morbidity and mortality has not decreased in the past two decades. In light of this, clinicians must remain abreast of the latest research and technological advances, and adopt those practices that improve outcomes. Continued critical appraisal of persistent racial and ethnic disparities may be useful in understanding and reversing current trends. Additionally we must continue to creatively develop instruments of quantifying those aspects of high quality prenatal care, which are unmeasurable. Furthermore, we must advocate on a local, state, and national level for improved services for our prenatal patients not just in the office and the hospital, but in their homes and communities. PMID- 12529896 TI - Well child and preventive care. AB - In summary, primary care providers play an extremely important role in providing preventive care. In the USA, the primary care physician is the major deliverer of well childcare. Numerous resources exist as a guide to the recommended content of health supervision visits. Despite the overall improvement in the health of children, areas that continue to challenge us, include childhood poverty and persistent health disparities among different ethnic groups. Although, solutions to these problems may well lie with political and social organizations that take a population-based approach to health care, each visit into the primary care provider's office provides an opportunity to review urgent concerns, update immunization status, address developmental and behavioral concerns and promote healthy lifestyles. PMID- 12529897 TI - Adolescent care: reducing risk and promoting resilience. AB - In summary adolescence is a crucial period in the chronology of health. Most lifelong health choices are decided at this stage and clinicians can help adolescents make healthy choices to ensure a safe, secure future. They also develop skills for the rest of their lives to cope with stress, take care of their bodies, and decide whom to spend time with in meaningful relationships. Preventive health for adolescence stems from the relationship the provider cultivates with the patient and the power that comes from that dynamic relationship to respond to patient needs and promote health throughout the life cycle. PMID- 12529898 TI - Prevention in adulthood: forging a doctor-patient partnership. AB - In this ever-changing health care system, Americans still retain confidence in their individual physicians [11]. This confidence represents an impressive fact when placed in the context of today's threats to the physician-patient relationship. In this chapter, we have explored these threats, principle among which are the appearance of the hospitalist, the information and technology revolution and the preeminence of managed care. We have further attempted to describe particular physician characteristics that may affect the quality of the physician-patient relationship. Individual physician awareness and characteristics seem to form the cornerstone for the quality of the relationship. We certainly believe in and stress the importance of shared mutual obligations in the good physician-patient relationship. Descriptors such as patients' trust and physicians' empathy enable meaningful relationships to be nurtured over time. A key component worthy of the primary care physician's consideration is that of longitudinal, continuity of care. In a study that examined patients' trust in their physicians, nearly three fourths of those surveyed were confident that their physicians would "put (the patient's) needs above all other considerations" [12]. Despite the limitations inherent in the modern American medical environment, it is still possible for physicians to nurture trusting, effective relationships with their patients and ultimately enhance health outcomes. PMID- 12529899 TI - Preventive care for the menopausal woman. AB - In summary, menopause represents a stage in a woman's life for the primary care physician to have an important impact on health [43]. Decision-making must weigh individual risk factors for health disease, osteoporosis and breast cancer. Hormone replacement therapy remains an important issue for the postmenopausal woman [44]. More evidence is needed to define the specific health benefits and risks of HRT, particularly as it relates to the prevention of CHD. A patient centered approach using a shared decision making model is most likely to result in improved patient satisfaction and desired health outcome. PMID- 12529900 TI - Preventive health care for older patients. AB - This chapter addresses key components of screening and preventive care for the older population. The older population is heterogeneous, ranging from the competent, active, well individual to the frail, demented individual. Certain preventive measures are important for all individuals such as counseling on exercise and screening for high blood pressure. However, universal cancer, cholesterol, or dementia screening may not be cost effective and beneficial in all older adults. These preventive measures should be guided by the individuals' circumstances including their life expectancy, co-morbid illnesses, functional capacity, and quality of life. Clinicians may be able to individualize preventive medicine decisions by stratifying their patients into well and frail using the guidelines we have provided. The goal of prevention and screening in older patients is to improve function and quality of life. Primary care physicians should facilitate discussion of preventive measures with their older patients as part of their ongoing health care. PMID- 12529901 TI - The role of nutrition and nutrients in the prevention of chronic diseases. AB - The nutritional factors that increase risk of heart disease, many cancers, and diabetes are identical. Controlling body weight, being physically active, limiting intake of saturated fats, and eating ample amount of fruits and vegetables can all reduce the risk for many different chronic diseases. Physicians have many opportunities to assist their patients in reducing their nutritional risk of chronic diseases. PMID- 12529902 TI - Tobacco smoking as a chronic disease: notes on prevention and treatment. AB - Tobacco use represents a rare confluence of interesting circumstances. Elements of inheritable risk combine with powerful neuropharmacology and a ubiquitous environmental exposure and result in an epidemic that claims over 430,000 lives and costs us over $100 billion annually. It is the single most important remediable public health problem in the United States. Most smokers want to quit smoking and a simple advice from a physician can increase the likelihood of doing so. Moreover, there are a number of pharmacologic and behavioral therapies that are proven to be effective in smoking cessation. Yet, there is an apparent reluctance among physicians to address smoking cessation, perhaps due to a sense of frustration or low self-efficacy. Physicians play an important role in smoking cessation, and intensive interventions are necessary to improve their participation and efficacy. Teaching practical smoking cessation techniques within medical school curricula, with an opportunity for standardized practice and self-evaluation, may be an effective strategy to improve physician practice in this area. Since most smokers try their first cigarette before the age of 18, and youth smoking is on the rise, targeted interventions aimed at preventing initiation and encouraging cessation of smoking among youth are needed. For all tobacco users, a better understanding of the pharmacology and physiology of nicotine addiction may translate into targeted and individualized treatment and prevention strategies, which may improve success rates dramatically. To better control this epidemic, and to meet the nation's public health goals for the year 2010 [145], local tobacco control interventions need to be multifaceted and well integrated into regional and national efforts [146]. Because of the physician's unique societal role with respect to tobacco, doctors may indeed find it possible to impact public opinion and significantly reduce the toll of tobacco by acting at the public health and public policy levels [147]. Those interested in engaging in the public health debate can do more than relay facts about tobacco and health. Involvement in tobacco-control issues provides the opportunity to impact the environmental influences promoting smoking among patients, and is likely to be synergistic with efforts to help smokers quit within the office. Physicians who take steps to engage in local public health initiatives are likely to magnify the effects of their efforts at the bedside [148, 149]. PMID- 12529903 TI - Immunizations in adulthood. AB - Vaccine preventable diseases continue to be a sizable burden to adults in the United States. Despite availability of safe and effective vaccines, immunization rates remain low. While various patient, clinician, and system factors can be barriers to adult vaccination, physicians recommendations to their patients to receive the vaccinations for which they are indicated is strongly related to a patient's receipt of these immunizations. PMID- 12529904 TI - Cardiovascular disease prevention. AB - In this chapter, we have reviewed many of the steps necessary for effective CHD risk reduction. The first step in the office setting is to assess the individual CHD risk. This combines the evaluation of current CHD or a "secondary risk equivalent" with the counting of risk factors and in many cases, the absolute risk calculation. The next steps are to consider each of the major modifiable risk factors (hypertension, dyslipidemia, diabetes mellitus, smoking status) to set goals for each and then work to achieve those goals through lifestyle changes and medication therapy. We reviewed each of these risk factors in detail and then turned to a discussion of emerging risk factors that may help "fine-tune" the risk assessment in some borderline cases. We also discussed additional non invasive testing that is available to the clinician to help refine the assessment of current burden of disease. Finally, we discuss some of the barriers that exist on both a global and local level to effective treatment of CHD risk factors. PMID- 12529905 TI - Cancer screening. AB - Table 3 provides a summary of key recommendations for each cancer site discussed in this chapter. One of the unifying principles of cancer screening is that every clinician or group practice needs to define an explicit screening policy. Resources must then be devoted to implementing this policy, evaluating adherence, and improving performance. PMID- 12529906 TI - Prevention in the year 2002: some news, some issues. AB - As we move into the new century, we continue to focus on ways to prevent disease, promote health, and prolong life. We have made enormous strides in some arenas, including deaths from infectious diseases caused by mass immunization, improved sanitation, better nutrition, heart disease caused by risk factor modification, and some cancers caused by smoking cessation and screening for early detection. Although we have made progress, many challenges remain. PMID- 12529907 TI - Occupational injury and safety research in the AIHA Journal. PMID- 12529908 TI - 2002 William P. Yant Award Lecture. Diesel particulate control strategies at some Australian underground coal mines. PMID- 12529909 TI - A method for estimation of bias and variability of continuous gas monitor data: application to carbon monoxide monitor accuracy. AB - A method is presented for the evaluation of the bias, variability, and accuracy of gas monitors. This method is based on using the parameters for the fitted response curves of the monitors. Thereby, variability between calibrations, between dates within each calibration period, and between different units can be evaluated at several different standard concentrations. By combining variability information with bias information, accuracy can be assessed. An example using carbon monoxide monitor data is provided. Although the most general statistical software required for these tasks is not available on a spreadsheet, when the same number of dates in a calibration period are evaluated for each monitor unit, the calculations can be done on a spreadsheet. An example of such calculations, together with the formulas needed for their implementation, is provided. In addition, the methods can be extended by use of appropriate statistical models and software to evaluate monitor trends within calibration periods, as well as consider the effects of other variables, such as humidity and temperature, on monitor variability and bias. PMID- 12529911 TI - Analytical model for evaluating lateral capture efficiencies in surface treatment tanks. AB - A pilot scale installation 1.8 m long and 1.6 m wide was designed to simulate a surface treatment tank incorporating a lateral capture system and a distribution network that permits the surface emission of tracer (SF6). The capture efficiency, ET, was determined as a function of the specific capture flow qc, m3/sm2, for three tank lengths, L, (1.2, 1.53, and 1.8 m) and several temperatures, T, from ambient to 95 degrees C, keeping the surrounding air velocity low (v < or = 0.1 m/sec). The data showed a satisfactory fit to a model that related capture efficiency with two exponential functions: the first dependent on the ratio capture velocity/air velocity in the vicinity of the tank, and the second on the ratio capture velocity/velocity of rising drafts provoked by the temperature of the tank. PMID- 12529910 TI - Statistical methods for establishing equivalency of a sampling device to the OSHA standard. AB - Occupational Safety and Health Administration (OSHA) regulations allow the use of an alternative sampling device for exposure monitoring provided the device has been demonstrated to be equivalent to the standard device. For example, the OSHA standard allows the use of an alternate cotton dust sampler that is equivalent to the Lumsden-Lynch vertical elutriator (VE); also, OSHA defines the accuracy of the monitoring device for measuring airborne chemicals such as benzene and sulfur dioxide. Typically, the OSHA criterion is that 90% of the readings of the sampling device should be within +/- 25% of the readings obtained by the standard device or within +/- 25% of the actual airborne chemical concentration. This article proposes two statistical tests for establishing that an alternative measuring device of airborne chemicals or dust is equivalent to the OSHA standard. The statistical tests are illustrated using an example. PMID- 12529912 TI - Photometer response determination based on aerosol physical characteristics. AB - Aerosol photometers often are calibrated against field measurements of total or respirable aerosol concentrations. However, the response of these instruments to a particular aerosol concentration level will change if there is a change in aerosol size distribution. To determine a predictable correction factor that relates photometer to gravimetric measurements of an aerosol, the authors performed an analysis of photometer response relative to particle size distribution, density, and refractive index. A series of trials was performed at different concentration levels using both Arizona road dust and ground corn dust. The correction factor was calculated from a knowledge of the instrument response pattern relative to particle size, as well as the ratios of actual dust density and refractive index relative to that of the dust used to calibrate the instruments. Results demonstrated that the calculated correction factors were within 10% of a correction factor predicted from the slope of a linear regression relating gravimetric measurements to the average of photometer readings taken during the sample period. PMID- 12529913 TI - Bending the tool and the effect on human performance: further investigation of a simulated wire-twisting task. AB - A previous experiment by the authors investigated the effect of the design of pliers on productivity. A simulated wire-twisting task performed at five heights between 10 inches above and 10 inches below elbow height formed the basis of the comparison. Pliers designed according to the American National Standard for long nose and long reach pliers were compared to bent-handled pliers designed at the Western Electric (WE) Co. The WE pliers, designed to reduce ulnar deviation during a wire-twisting task, resulted in productivity decrements between 1.4 and 17% across the five heights studied. The current experiment was a continuation of that study, adding orientation of the workplace in the sagittal plane as an independent variable. Wrist deviation in two planes were additional dependent measures, and the experiment was performed by female subjects with a mean age of 48.1 years. The results did not indicate a significant difference between productivity levels for the two pliers designs. Work height and work orientation, as well as their interaction, tended to have the strongest effects on productivity and wrist deviation. In general, productivity was inversely related to measures of discomfort and postural deviation. PMID- 12529914 TI - Maximal acceptable torques of highly repetitive screw driving, ulnar deviation, and handgrip tasks for 7-hour workdays. AB - This study represents a continuation of a series of psychophysical studies on repetitive motions of the wrist and hand conducted at the Liberty Mutual Research Center for Safety and Health. The purpose of the study was to quantify maximum acceptable torques of six motions performed on separate days but within the context of the same experiment. The six motions were screw-driving clockwise with a 31-mm handle, a 40-mm handle, and a 39-mm yoke handle; screw-driving counterclockwise with a 31-mm handle; ulnar deviation with a power grip (similar to knife cutting), and a handgrip task (similar to a pliers task). A psychophysical methodology was used in which the subject adjusted the resistance on the handle, and the experimenter manipulated or controlled all other variables. Ten subjects performed the six tasks at repetition rates of 15, 20, and 25 motions per minute. Subjects performed the tasks for 7 hours per day, 5 days per week, for 4 weeks. The subjects were instructed to work as if they were on an incentive basis, getting paid for the amount of work they performed. Symptoms were recorded by the subjects during the last 5 min of each hour. The results revealed that mean maximum acceptable torques ranged from 0.33 to 0.65 Nm for screw driving, 1.08 to 1.13 Nm for ulnar deviation, and 4.80 to 4.85 Nm for the handgrip task. These values represent 14 to 24% (median of 17%) of maximum isometric torque depending on the frequency and motion. A table of maximum acceptable torques and forces of the six motions is presented for application in the field. PMID- 12529915 TI - Crab allergen exposures aboard five crab-processing vessels. AB - Aerosolized crab allergens are suspected etiologic agents for asthma among crab processing workers. The objectives of this study were to characterize crab allergen concentrations and respiratory symptom prevalence among processing workers aboard crab-processing vessels. A cross-sectional survey of five crab processing vessels was conducted near Dutch Harbor, Alaska. Crab allergen concentrations were quantified during specific work activities with 25 personal air samples collected on polytetrafluoroethylene filters and analyzed by a competitive IgE immunoassay technique. Two standardized respiratory questionnaires were used to assess respiratory symptoms suggestive of bronchitis or asthma in 82 workers. Aerosolized crab allergen concentrations ranged from 79 ng/m3 to 21,093 ng/m3 (mean = 2797 ng/m3, SD = 4576 ng/m3). The highest concentrations were measured at butchering/degilling work stations, which were combined on the smallest vessel. A significant percentage of workers reported development of respiratory symptoms during the crab-processing season. Cough developed in 28% of workers, phlegm in 11% of workers, and wheeze and other asthma-like symptoms developed in 4% of workers. Despite variations in crab allergen levels, respiratory symptom prevalence was similar across all job categories. Substantial concentrations of crab allergen exposure were measured, as well as the potential for wide variability in exposure during crab processing aboard vessels. The high prevalence of reported respiratory symptoms across all job categories suggests potential adverse respiratory effects that should be further characterized by prospective studies using pulmonary function and serology testing, and rigorous exposure characterization. PMID- 12529916 TI - Diesel particulate matter exposure to railroad train crews. AB - Exposure assessments were conducted aboard diesel locomotives. Results were evaluated to determine variables that affect exposure to DPM (diesel particulate matter) and to assess use of EC (elemental carbon) and OC (organic carbon) as surrogates for DPM. National Institute for Occupational Safety and Health Method 5040 was used for collection and analysis of samples in locomotives and in nonrailroad settings. The level of EC, but not OC, in locomotives was found to be significantly affected by position of exhaust stacks and windows. EC ranged from < 1 to 45 micrograms/m3 with a geometric mean (GM) of 3.7 micrograms/m3 and OC ranged from 4 to 4570 micrograms/m3 with a GM of 36.3 micrograms/m3. Background measurements of EC ranged from < 1 to 8 micrograms/m3 and OC levels were 4 to 84 micrograms/m3. This study confirms that train crew exposure to DPM is much lower than exposures for miners, is comparable to background urban exposures, and is lower than but comparable to exposures for truck drivers. It also indicates that EC levels are highly predictive of diesel exhaust exposure whereas OC levels are not, and that open windows and exhaust stack(s) in front of the locomotive cab have a significant effect on EC. PMID- 12529917 TI - Task-based assessment of occupational vibration and noise exposures in forestry workers. AB - Forty-two noise exposures and 164 whole-body (WBV) and hand-arm (HAV) vibration exposures were collected from 43 forestry workers in six trades employed by two forestry companies. Data were collected on 10 days over 8 weeks during various felling, logging, and log handling operations. Up to 5 volunteers were monitored for noise and vibration daily using datalogging noise dosimeters, which provided daily time-weighted averages (TWAs) and 1-min averages; and a precision sound level meter equipped to measure human vibration, which provided triaxial HAV and WBV event-weighted averages (AEQS). Workers completed a short questionnaire throughout the workday detailing the timing and number of tasks performed and equipment used. Substantial overexposures to noise and vibration were seen; for example, 60% of Occupational Safety and Health Administration (OSHA) TWAs and 83% of National Institute for Occupational Safety and Health (NIOSH) noise TWAs exceeded 85 dBA, 33-53% of the axis-specific HAV AEQS exceeded the 8-hour American Conference of Governmental Industrial Hygienists' HAV threshold limit value, and 34% of all summary weighted WBV AEQS exceeded the Commission of the European Communities' 8-hour exposure limit. The mean for 99 WBV summary weighted AEQ was 3.53 +/- 7.12 m/sec2, whereas the mean for 65 HAV summary weighted AEQ was 5.45 +/- 5.25 m/sec2. The mean OSHA TWA was 86.1 +/- 6.2 dBA, whereas the mean NIOSH TWA was 90.2 +/- 5.1 dBA. The task and tool with the highest exposure levels were unbelling chokers on landings and chain saws (noise), log processing and frontend loaders (WBV), and notching stumps and chain saws (HAV). An internal validation substudy indicated excellent agreement between worker-reported and researcher-documented tasks and tools. PMID- 12529918 TI - Evaluation of worker exposure to asphalt paving fumes using traditional and nontraditional techniques. AB - Forty-five workers at 11 paving sites across the United States were evaluated for exposure to paving asphalt (bitumen) fumes. Traditional measures of exposure such as total particulate matter (TPM) and benzene soluble matter (BSM) were monitored. In addition, total organic matter (TOM), which includes both the BSM residue and the more volatile components that pass through the filter and are collected on sorption material, was quantified and further characterized using a gas chromatography technique and a recently developed fluorescence test. The latter method, which indirectly estimates the content of four- to six-ring polycyclic aromatic compounds, is used as a predictor of carcinogenicity. The correlation between fluorescence emission intensity and carcinogenicity for 36 laboratory generated fume fractions, as measured in a mouse skin-painting bioassay, was then used to estimate the carcinogenic potential of worker monitoring samples. Emission levels, and therefore predicted carcinogenicity, for these samples were at least 17-fold below the value corresponding to a minimal carcinogenic effect. This result was consistent with more extensive chemical analysis (using gas chromatography/mass spectrometry) of two of the samples, which showed the predominant constituents to be alkanes, monocycloparaffins, alkyl-benzenes, alkyl-naphthalenes, and alkyl-benzothiophenes. The geometric mean exposures for all worker studies were 0.21 mg/m3 (TPM), 0.06 mg/m3 (BSM), and 1.23 mg/m3 (TOM). PMID- 12529919 TI - Should the threshold limit value for power frequency (60 Hz) magnetic fields be changed? Perceptions among scientists and other risk experts. AB - Over the past few years several studies have raised questions about childhood cancer and exposure to electric and magnetic fields (EMFs). Reports by the National Academy of Sciences and the National Cancer Institute concluded that there is no clear, convincing evidence that exposure to electric power lines and electric power appliances is a threat to human health, whereas the National Institute of Environmental Health Sciences concluded that there is weak evidence that EMF exposure may be a leukemia hazard. Based on the conflicting agency reports on EMF, what do experts in the field of bioelectromagnetics believe regarding the effectiveness of the American Conference of Governmental Industrial Hygienists (ACGIH) guidelines? Surveys were sent to 163 participants at the 1997 annual Department of Energy Contractors meeting on Electric and Magnetic Fields. Approximately one-half (49%, n = 81) of the participants returned the survey. Utility workers demonstrated greater levels of agreement with the current TLV standard in comparison to government, university, and other private sector workers. Utility workers were less likely to perceive that ACGIH guidelines should be changed to be made more strict. In addition, workers who had worked at their present job for 10 years or more indicated less agreement with the acceptability of ACGIH guidelines to protect workers' health. The scientists and other risk experts in this study illustrate the importance of how type of industry may influence perceptions of TLV effectiveness. PMID- 12529920 TI - Percutaneous absorption of inorganic lead compounds. AB - The objective of this study was to determine percutaneous absorption of lead compounds, including lead sulfate, lead oxide, lead powder, and lead stearate. The lead content on the skin surface of 10 lead-battery workers was measured by the method of skin stripping, and urinary lead content of rats was measured with epicutaneous application of four lead compounds: lead sulfate, lead oxide, lead powder, and lead stearate. There were significant amounts of lead on the 9th and 10th skin strippings of the dorsal hand and the back of lead workers. The amount of lead on the dorsal hand was significantly correlated with the amount in the blood (n = 10, r 2 = 0.66, p < 0.05, linear regression). In rats, after lead compounds were applied for 12 days, total lead amount in urine significantly increased to 146.0 +/- 6.4 ng (SD) for lead stearate, 123.1 +/- 7.2 ng for lead sulfate, 115.9 +/- 5.3 ng for lead oxide, 47.8 +/- 6.9 ng for lead powder, and 10.3 ng for the control, which indicated significant skin absorption. It was concluded that significant amounts of inorganic lead compounds can be absorbed through the skin, and skin protection in lead-working or any contaminated environment should be carefully considered. PMID- 12529921 TI - Exposure to formaldehyde among animal health students. AB - The objective of the present study was to evaluate the exposure to formaldehyde in 2 groups each with 18 students in animal health technology from two different training centers (TC) during a 3-hour weekly laboratory session in biology. Personal sampling during the session was done with passive bubblers for formaldehyde. The analysis of formaldehyde was done by visible absorption spectrometry according to NIOSH method 3500. The students in TC 1 were exposed to formaldehyde levels ranging from less than 0.11 to 0.76 mg/m3 with an average at 0.25 mg/m3 during the 3-hour biology laboratory. The students in TC 2 were exposed to higher concentrations of formaldehyde ranging from 0.26 to 1.2 mg/m3 with an average at 0.632 mg/m3. The results of the survey indicate that the students in both centers were exposed to a notable amount of formaldehyde vapor, at a level that is above the threshold limit value. PMID- 12529922 TI - Exposure of miners to diesel exhaust particulates in underground nonmetal mines. AB - A study was initiated to examine worker exposures in seven underground nonmetal mines and to examine the precision of the National Institute for Occupational Safety and Health (NIOSH) 5040 sampling and analytical method for diesel exhaust that has recently been adopted for compliance monitoring by the Mine Safety and Health Administration (MSHA). Approximately 1000 air samples using cyclones were taken on workers and in areas throughout the mines. Results indicated that worker exposures were consistently above the MSHA final limit of 160 micrograms/m3 (time weighted average; TWA) for total carbon as determined by the NIOSH 5040 method and greater than the proposed American Conference of Governmental Industrial Hygienists TLV limit of 20 micrograms/m3 (TWA) for elemental carbon. A number of difficulties were documented when sampling for diesel exhaust using organic carbon: high and variable blank values from filters, a high variability (+/- 20%) from duplicate punches from the same sampling filter, a consistent positive interference (+26%) when open-faced monitors were sampled side-by-side with cyclones, poor correlation (r 2 = 0.38) to elemental carbon levels, and an interference from limestone that could not be adequately corrected by acid washing of filters. The sampling and analytical precision (relative standard deviation) was approximately 11% for elemental carbon, 17% for organic carbon, and 11% for total carbon. An hypothesis is presented and supported with data that gaseous organic carbon constituents of diesel exhaust adsorb onto not only the submicron elemental carbon particles found in diesel exhaust, but also mining ore dusts. Such mining dusts are mostly nonrespirable and should not be considered equivalent to submicron diesel particulates in their potential for adverse pulmonary effects. It is recommended that size-selective sampling be employed, rather than open-faced monitoring, when using the NIOSH 5040 method. PMID- 12529923 TI - Determination of carbon disulfide at the workplace by sampling on charcoal tubes- problems and solutions. AB - The aim of the study was to check the reliability and comparability of different analytical methods for ambient monitoring of carbon disulfide (CS2). A stationary sampling system, consisting of a charcoal sampling tube and pump, and two personal sampling systems, consisting of a charcoal sampling tube and a portable pump and of a diffusive charcoal sampler have been compared. The limits of quantification, within-series precision, between-series precision, recovery, and comparability of the methods were investigated. For passive sampling the recovery was determined by three different techniques. For a sampling time of 6 hours the limit of quantification was 0.2 ppm for the personal sampling and 0.01 ppm for the stationary sampling system. The within-series precision was between 5 and 8%. For personal sampling the between-series precision was between 9 and 12% using a passive sampler. The recovery ranged between 45 and 85% depending on the quotient of eluent volume and charcoal mass. The comparison of the two personal sampling methods in a field study using linear regression demonstrated with statistical certainty an excellent concordance of the methods. The study shows that particularly the method for determining air levels of carbon disulfide by passive sampling is associated with high systematic errors. If these errors are unknown or ignored they will result in highly underestimated exposure data for carbon disulfide. PMID- 12529924 TI - ELPAT program report: background and current status. PMID- 12529925 TI - Circulating amyloid-beta peptide crosses the blood-brain barrier in aged monkeys and contributes to Alzheimer's disease lesions. AB - 1. We studied cerebrovascular sequestration and blood-brain barrier (BBB) permeability to [125I]- or [123I]-labeled amyloid-beta peptides (A beta) in aged rhesus and aged squirrel monkey, the nonhuman primate models of cerebral beta amyloidosis and cerebrovascular amyloid angiopathy (CAA), respectively. 2. In aged rhesus, the half-time of elimination of [125I]A beta 1-40, t1/2e, was faster by 1.34 h, the systemic clearance, Clss, increased by 4.21 ml/min/kg and the mean residence time of intact peptide in the circulation shortened by 2 h. 3. Cerebrovascular sequestration of [125I]A beta 1-40 was significant in aged squirrel monkey (20.8 ml/g x 10(2)), but undetectable in the rhesus. 4. The permeability surface area product, PS, for [14C]inulin was low in both species (0.11-0.18 ml/g/s x 10(6)) indicating an intact barrier. 5. The BBB permeability to A beta 1-40 was 34.8- and 13.7-fold higher than for [14C]inulin in aged squirrel and rhesus, respectively, suggesting a specialized A beta transport across the BBB. 6. The single photon computed emission tomography studies confirmed a saturable [123I]A beta 1-40 transport at the BBB in primates (Km = 40 nM). 7. Brain autoradiographic analysis of [125I]A beta 1-42 or [125I]A beta 1-40 after intracarotid infusions of radiotracers confirmed co-localization of the signal with A beta-immunoreactive plaques in rhesus monkeys. 8. Metabolism of [125I]A beta 1-40 in brain and plasma was slower in aged squirrel compared to aged rhesus, by 2.9- and 2.6-fold, respectively. 9. Thus, transport of circulating A beta across the BBB contributes to brain parenchymal accumulation of amyloid in aged nonhuman primates. Negligible capillary binding, rapid systemic and brain degradation, and accelerated body elimination of blood-borne A beta, may prevent the development of CAA in rhesus in contrast to squirrel monkeys. PMID- 12529926 TI - Lymphocyte migration into the central nervous system: implication of ICAM-1 signalling at the blood-brain barrier. AB - Lymphocyte recruitment to the central nervous system (CNS) is a critical step in the pathogenesis of diseases such as multiple sclerosis (MS), meningitis and posterior uveitis. The principle sequential stages that control lymphocyte emigration from the blood have been widely reported, but only recently has attention been directed towards the role of the vascular endothelium in actively supporting transvascular migration. It has now been shown that adhesion molecules, particularly those of the immunoglobulin super family (e.g. ICAM-1, VCAM-1 and PECAM-1), not only act as ligands for leucocyte receptors but can also serve as signal transducers. Engagement of these receptors initiates endothelial signalling cascades that result in downstream effector mechanisms which in turn influence the progression of neuroinflammation. In particular, it has been shown that ICAM-1-mediated signalling in brain endothelial cells is a crucial regulatory step in the process of lymphocyte migration through the blood-brain barrier and as such represents an additional phase in the multistep paradigm of leucocyte recruitment. In this article we review current understanding of endothelial cell ICAM-1 signalling and discuss the importance of these findings in relation to leucocyte trafficking to the CNS. PMID- 12529927 TI - Tight junctions of the blood-brain barrier: development, composition and regulation. AB - 1. The blood-brain barrier is essential for the maintenance and regulation of the neural microenvironment. The main characteristic features of blood-brain barrier endothelial cells are an extremely low rate of transcytotic vesicles and a restrictive paracellular diffusion barrier. 2. Endothelial blood-brain barrier tight junctions differ from epithelial tight junctions, not only by distinct morphological and molecular properties, but also by the fact that endothelial tight junctions are more sensitive to microenvironmental than epithelial factors. 3. Many ubiquitous molecular tight junction components have been identified and characterized including claudins, occludin, ZO-1, ZO-2, ZO-3, cingulin and 7H6. Signaling pathways involved in tight junction regulation include G-proteins, serine-, threonine- and tyrosine-kinases, extra and intracellular calcium levels, cAMP levels, proteases and cytokines. Common to most of these pathways is the modulation of cytoskeletal elements and the connection of tight junction transmembrane molecules to the cytoskeleton. Additionally, crosstalk between components of the tight junction- and the cadherin-catenin system of the adherens junction suggests a close functional interdependence of the two cell-cell contact systems. 4. Important new molecular aspects of tight junction regulation were recently elucidated. This review provides an integration of these new results. PMID- 12529928 TI - Drug transport to the brain: key roles for the efflux pump P-glycoprotein in the blood-brain barrier. AB - 1. The blood-brain barrier (BBB) contributes to brain homeostastis and fulfills a protective function by controlling the access of solutes and toxic substances to the central nervous system (CNS). The efflux transporter P-glycoprotein (P-gp) is a key element of the molecular machinery that confers special permeability properties to the BBB. 2. P-gp, which was initially recognized for its ability to expel anticancer drugs from multidrug-resistant cancer cells, is strongly expressed in brain capillaries. Its expression in the BBB limits the accumulation of many hydrophobic molecules and potentially toxic substances in the brain. 3. The purpose of this review is to summarize the current state of knowledge about the expression of P-gp, its cellular localization as well as its possible functions in the BBB. PMID- 12529929 TI - Drug delivery to brain via the blood-brain barrier. AB - 1. Many neurodegenerative diseases, cancer and infections of the brain become more prevalent as populations become older. Despite major advances in neuroscience, the blood-brain barrier (BBB) ensures that many potential therapeutic cannot reach the central nervous system (CNS). The BBB is formed by the complex tight junctions between the endothelial cells of the brain capillaries and their low endocytic activity. This results in the capillary wall that behaves as a continuous lipid bilayer and prevents the passage of polar and lipid-insoluble substances. It is, therefore, the major obstacle to drugs that may combat diseases affecting the CNS. 2. Several strategies for delivering drugs to the CNS have been developed. These enhance the capacity of therapeutic molecules to cross the BBB by modifying the drug itself, or by coupling it to a vector for receptor-mediated or adsorption-mediated transcytosis. 3. The current challenge is to develop drug-delivery systems that ensure that drugs cross the BBB in a safe and effective manner. This review focuses on the strategies developed to enhance drug delivery across the BBB. PMID- 12529931 TI - Molecular pharmacology of class II G protein-coupled receptors. Introduction. PMID- 12529930 TI - The use of in vitro cell culture models for mechanistic studies and as permeability screens for the blood-brain barrier in the pharmaceutical industry- background and current status in the drug discovery process. AB - Successful drug delivery to the central nervous system (CNS) is highly dependent on DMPK as well as physicochemical properties and it is therefore important to characterise these properties and take them into account when designing chemical lead series that act at CNS targets. Since the drug discovery/development process is becoming increasingly focused on reducing the time required to enter molecules into the market, industrial DMPK scientists have emerged from their traditional supportive role in drug development to provide valuable support in the drug discovery process, using novel methods to meet the demands of combinatorial chemistry and bioscience groups. PMID- 12529932 TI - VPAC receptors for VIP and PACAP. AB - VIP and PACAP are two prominent neuropeptides that share two common G protein coupled receptors, VPAC1 and VPAC2, while PACAP has an additional specific receptor, PAC1. This article reviews the present knowledge regarding various aspects of VPAC receptors including: 1) receptor specificity toward natural VIP related peptides and pharmacology of synthetic agonists or antagonists; 2) genomic organization and chromosomal localization; 3) signaling and established or putative interactions with G proteins or accessory proteins such as RAMPs or PDZ-containing proteins; 4) molecular basis of ligand-receptor interaction as determined by site-directed mutagenesis, construction of receptor chimeras, and structural modeling; 5) constitutively active receptor mutants; 6) short-term (desensitization, internalization, phosphorylation) and long-term (transcription) regulations and transgenic models; 7) receptor polymorphisms. PMID- 12529933 TI - Growth hormone releasing hormone receptor. AB - Growth hormone releasing hormone receptor (GHRH-R) is a class II G protein coupled receptor required for normal growth hormone (GH) synthesis and release from the pituitary, and for the normal growth and proliferation of somatotrophs within the pituitary. Mutations of this receptor in mouse and human are associated with GH deficiency, short stature, and pituitary hypoplasia. This signaling system plays important roles in growth and development, metabolism of muscle and fat, and, possibly, the aging process. Among topics touched on in this article are the receptor gene's organization and promoter regulation, receptor signaling pathways, and receptor domains involved in interaction with ligand. Also discussed are alternative mRNA splicing and proposed functions of these splice variants as dominant negative inhibitors of wild-type receptor and in tumor proliferation. Homology among GHRH-Rs from different species is shown and a map of receptor mutations reported to effect function is presented. PMID- 12529934 TI - Pharmacology and biology of corticotropin-releasing factor (CRF) receptors. AB - The biology of corticotropin-releasing factor (CRF) finds increasing interest in the scientific community because of the neuromodulatory actions of CRF on brain functions such as learning, anxiety, feeding, and locomotion. Additional actions on immunumodulation and apoptosis have recently been discovered. All actions of CRF are mediated by G protein-coupled receptors, which trigger different, sometimes opposite actions in different regions of the central nervous system. The CRF system exhibits considerable plasticity by the involvement of numerous different ligands, splice variants, and transductional couplings. The generation of multiple splice variants is facilitated by the intron exon structure of the CRF receptor genes. PMID- 12529935 TI - Structure-function of the glucagon receptor family of G protein-coupled receptors: the glucagon, GIP, GLP-1, and GLP-2 receptors. AB - The glucagon-like peptides include glucagon, GLP-1, and GLP-2, and exert diverse actions on nutrient intake, gastrointestinal motility, islet hormone secretion, cell proliferation and apoptosis, nutrient absorption, and nutrient assimilation. GIP, a related member of the glucagon peptide superfamily, also regulates nutrient disposal via stimulation of insulin secretion. The actions of these peptides are mediated by distinct members of the glucagon receptor superfamily of G protein-coupled receptors. These receptors exhibit unique patterns of tissue specific expression, exhibit considerable amino acid sequence identity, and share similar structural and functional properties with respect to ligand binding and signal transduction. This article provides an overview of the biology of these receptors with an emphasis on understanding the unique actions of glucagon related peptides through studies of the biology of their cognate receptors. PMID- 12529936 TI - Molecular pharmacology of the secretin receptor. AB - The secretin receptor was the first member of the Class II family of G protein coupled receptors to be cloned. It is prototypic of this family in its structure, function, and regulation. The extended amino-terminal tail domain includes a series of six conserved Cys residues that contribute three intradomain disulfide bonds. This region of the receptor has been shown by mutagenesis and photo affinity labeling to be particularly important in secretin binding and stimulation of signaling activity. There is clear evidence for the direct interaction of the natural agonist peptide with this receptor domain. Mutagenesis has also identified important contributions of extracellular loop domains, although their specific roles remain unclear. This receptor is regulated by agonist-stimulated phosphorylation and internalization, with details dependent on the cellular environment. PMID- 12529937 TI - Receptors for calcitonin gene-related peptide, adrenomedullin, and amylin: the contributions of novel receptor-activity-modifying proteins. AB - The discovery of receptor-activity-modifying proteins (RAMP) revealed a new principle for the function of G protein-coupled receptors. The initially orphan calcitonin receptor-like receptor (CRLR) was identified as a CGRP receptor when coexpressed with RAMP1. The same receptor is specific for adrenomedullin (ADM) in the presence of RAMP2. Calcitonin receptors (CTR) with 60% homology to the CRLR predominantly recognize calcitonin in the absence of RAMP. An amylin/CGRP receptor was recognized when a calcitonin receptor (CTR) was coexpressed with RAMP1. In the presence of RAMP3, the CTR only interacts with amylin. Noncovalent association of the RAMP with the CRLR or the CTR reveals heterodimeric RAMP/receptor complexes at the cell surface. Thus, two Class II G protein-coupled receptors, the CRLR and CTR, associate with three RAMP to form high affinity receptors for CGRP, ADM, or amylin. Here, the molecular composition and the functional properties of these receptors is reviewed. PMID- 12529938 TI - The parathyroid hormone 2 (PTH2) receptor. AB - The human PTH2 receptor binds and is activated at high potency by PTH and by the recently discovered peptide tuberoinfundibular peptide of 39 residues (TIP39). Rat and zebrafish PTH2 receptors are more weakly activated by PTH, suggesting that TIP39 may be the natural ligand for the PTH2 receptor. Unlike the PTH1 receptor, the PTH2 receptor interacts extremely weakly with parathyroid hormone related peptide (PTHrP). The PTH2 receptor is strongly coupled to stimulation of cAMP accumulation, and more weakly, in a cell-specific manner to increases in intracellular calcium concentration. A variety of evidence supports the general model of receptor amino terminal sequences binding ligand carboxyl terminal sequences with high affinity, and ligand amino terminal sequences activating the receptor through interaction with the "juxtamembrane" portion of the receptor. The receptor is present at greatest levels in the nervous system. It is expressed in scattered cells in the cerebral cortex and basal ganglia and at relatively high abundance in the septum, midline thalamic nuclei, several hypothalamic nuclei, and the dorsal horn of the spinal cord. Peripherally, expression in pancreatic islet somatostatin cells is most dramatic. Functional hypotheses based on the receptor's distribution are being tested. Recent data support involvement in hypothalamic releasing-factor secretion and pain. PMID- 12529939 TI - Parathyroid hormone 1 receptor: insights into structure and function. AB - The parathyroid hormone (PTH) and parathyroid hormone type 1 receptor (PTH1-Rc) are major players in regulating blood calcium homeostasis. PTH1-Rc is a G protein coupled receptor (GPCR) and it is also activated by PTH-related protein (PTHrP), which has potent effects in several target tissues including bone, via endocrine, paracrine, and autocrine mechanisms in prenatal and adult stages of life, and in several diseases. Another layer of complexity is added by the recently discovered PTH2-Rc subtype displaying unique tissue distribution and ligand specificity, and the potential presence in mammals of a third receptor subtype. Understanding of the mechanisms regulating PTH1-Rc gene expression, receptor desensitization, endocytosis, recycling, and intracellular signaling is advancing and provides insights to understand PTH1-Rc's role in both normal and pathophysiology. In addition, development of an experimentally based model of the PTH-PTH1-Rc complex by combining photoaffinity crosslinking, mutagenesis, conformational analysis, and molecular modeling sheds light, at the atomic resolution, on the molecular mechanisms involved in ligand binding, receptor activation, and coupling to intracellular effectors. Taken together, these lines of investigation offer not only a better understanding of GPCR mechanisms of action in general, but also contribute to the treatment of PTH/PTHrP-PTH1-Rc-related disease states and to the rational development of novel mechanism-based drugs to treat them. PMID- 12529940 TI - Molecular pharmacology of the calcitonin receptor. AB - The peptide hormone calcitonin is widely used therapeutically in the treatment of bone disorders such as Paget's disease, osteoporosis, and the hypercalcemia of some malignancies. However, emerging evidence suggests the actions of calcitonin via its G protein-coupled receptor, the calcitonin receptor, may not be limited to bone. Calcitonin receptors have also been identified in the central nervous system, testes, skeletal muscle, lymphocytes, and the placenta. We are now becoming aware that the complexity of the calcitonin response mediated by the calcitonin receptor can be influenced by accessory proteins, receptor isoforms, genetic polymorphisms, developmental and/or transcriptional regulation, feedback inhibition, and the specific cellular or tissue background. This article discusses what is known about the molecular and pharmacological actions of the calcitonin receptor and highlights areas of current research. PMID- 12529941 TI - [The new Ugeskrift--current, debate creating and scientific]. PMID- 12529942 TI - [Hypodynamic respiratory insufficiency. Diagnostic investigation]. AB - Patients with restrictive lung disease, owing to respiratory muscle dysfunction, have no parenchymal involvement. Their vital capacity (VC) and total lung capacity (TLC) are reduced to less than 50% and can lead to pneumonia and nocturnal hypercapnia and hypoxia. Their diffusion capacity is normal. With maximal static mouth pressure (Pimax) < 80 cm H2O and/or Pemax < 100 cm H2O, patients are referred to the national centres. Here, inspiratory muscular insufficiency is confirmed by sniff nasal inspiratory pressure and oesophageal pressure < 70 cm H2O. Expiratory muscular insufficiency is confirmed by a cough peak flow < 3-4 L/sec. and cough gastric pressure < 100 cm H2O. Sleep studies reveal nocturnal hypoventilation. Phrenic nerve stimulation is to be introduced in the diagnostic approach. Twitch mouth or oesophageal pressure < 10 cm H2O and twitch gastric pressure < 7 cm H2O are pathognomonic for neuromuscular respiratory insufficiency. PMID- 12529943 TI - [Surveillance and prevention of nosocomial Legionella pneumonia]. AB - Statens Serum Institut surveys the occurrence of nosocomial pneumonia caused by Legionella. The rate is low compared to other nosocomial infections but carries a high mortality. Verification of the diagnosis and acquisition (nosocomial or community acquired infection) is carried out in each suspected case. The criteria used for classification are described. Preventive measures include protection of susceptible patients as well as maintenance of the hot-water supply providing a minimum temperature of 50 degrees C at any hot tap. Technical solutions to reduce the concentration of Legionella in hospital hot-water supply systems are listed, and a guideline for the control and surveillance of the occurrence is suggested. PMID- 12529944 TI - [Interventions for prevention of obesity targeting mostly on non-obese children . Analysis of a Cochrane Review]. PMID- 12529945 TI - [Accelerated recovery program after hip fracture surgery]. AB - INTRODUCTION: A multimodal approach to minimise the effect of the surgical stress response can reduce complications and hospital stay after abdominal surgery and hip arthroplasty. The aim of the study was to assess the results of a well defined rehabilitation programme after hip fracture. MATERIAL AND METHODS: In an open intervention study, we entered 200 consecutive patients with hip fracture allowing full weight-bearing after operative treatment. The effect of a revised, optimised perioperative care programme with continuous epidural analgesia, early oral nutrition, oxygen supplementation, restricted volume and transfusion therapy, and intensive physiotherapy and mobilisation was assessed (n = 100) and compared with the conventional perioperative treatment programme before the intervention (n = 100). The median age was 82 (56-96) years in the control group and 82 (63-101) years in the accelerated multimodal perioperative treatment group. RESULTS: The median hospital stay was reduced from 21 (range 1-162, mean 32) to 11 (range 1-100, mean 17) days. The total use of days in hospital was reduced from 3211 to 1667. There were fewer complications, whereas the need for home care after discharge was unchanged. CONCLUSION: An accelerated clinical pathway with focus on pain relief, oral nutrition, and rehabilitation may reduce hospital stay and improve recovery after hip fracture. PMID- 12529946 TI - [Initial antibiotic therapy of purulent meningitis in adults. An investigation of practice patterns at Danish hospital department in 2000]. AB - INTRODUCTION: A previous report demonstrated profound variation in the recommended empirical antibiotic therapy for adult purulent meningitis in Denmark. At present, the only existing "official" recommendation (from the Danish Medical Association) is penicillin monotherapy for all adults, irrespective of age, as the prevalence of penicillin-resistant pneumococci is less than 3%. MATERIAL AND METHODS: A questionnaire concerning empirical antibiotic therapy and the treatment of specific microorganisms was sent to the 125 departments of internal medicine, infectious diseases, clinical microbiology, neurosurgery, otorhinolaryngology, and neurology in 2001. RESULTS: Consensus was not found in the 93% who replied, neither within or between the medical specialties nor within or between the counties. The departments of medicine and clinical microbiology were evenly distributed between two strategies: 1) a third-generation cephalosporin plus a penicillin, or 2) penicillin monotherapy, in supplemented in few departments with an aminoglycoside. For Haemophilus influenzae meningitis, 20% of the departments used ampicillin, whereas the majority preferred ceftriaxone. DISCUSSION: Consensus on empirical antibiotic treatment of purulent meningitis in adults had still not been reached in Denmark in the year 2000, and regimens other than that recommended by the Danish Medical Association were used. Complete and updated information is lacking on the resistance-patterns of bacteria inducing meningitis, and a complete national clinical microbiological database could form the basis for an evidence-based national consensus. PMID- 12529947 TI - ["The body-consciousness in school"--a back pain-school]. AB - INTRODUCTION: The aim of the present study was to demonstrate the extent of back pain in a school population and to evaluate if it was possible to minimize the number of complaints by increasing the body-consciousness in school children and education of pupils, teachers, and parents. MATERIAL AND METHODS: A prospective, controlled interventional study in two comparable schools with pupils from pre school up to and including ninth level. All pupils and their parents completed a questionnaire before and after the intervention. RESULTS: The share of pupils with back pain increased in the older age groups, whereas the frequency of physical activity decreased. Pupils who disliked going to school had more frequently back pain than pupils who liked going to school, and pupils who liked going to school were more physically active. There was a correlation between the back pain of the pupils and that of their parents. The intervention did not have any effect on the back pain of the pupils or their parents. DISCUSSION: There was no significant effect of the intervention, which might be explained by the relatively short time of implementation and unexpected practical problems at the school. The base-line values showed a correlation between age and back pain and between back pain and "do not like going to school". The frequency of physical activity decreased in the older age groups. Physically active pupils liked best going to school. There is a uniformity in the complaints of the parents and children and their reaction to the symptoms. PMID- 12529948 TI - [Causes of mental retardation in children of immigrant background. A registry study of the occurrence of consanguinity among parents of mentally retarded children at the Center for Handicapped, Glostrup hospital, county of Copenhagen]. AB - INTRODUCTION: In the County of Copenhagen the treatment of mentally retarded (MR) children is centralised in the Center for Handicapped (CFH). The aim of this study was to elucidate whether the frequency of MR is higher in children from ethnic minorities than in the background population and whether an increase could be ascribed to inbreeding. MATERIAL AND METHODS: The medical records of children with a non-Danish family name and where both parents were from countries other than Denmark were studied. RESULTS: A significant increase in the frequency of MR were found in the children from ethnic minorities: 16.6% of the children seen at CFH had an ethnic background compared to 14% in the background population, making a surplus of 18.5% more than expected. When a family was inbred other family members had Mr in 43.0%, whereas more family members with MR were found in only 14.2% of the families with no inbreeding. In the consanguineous families we found possible aetiological diagnoses in 21.5% of the children compared to 49% in the non-consanguineous families. In the entire group of children from ethnic minorities, we found Down's syndrome in 3.2% compared to 10.7% in Danish children with MR. DISCUSSION: There is a surplus of MR in children from ethnic minorities in the County of Copenhagen. The causes are not known, nor are aetiological factors for MR for a great part of the children. Consanguinity is likely to be a risk factor for MR, especially if it is already present in the family. This conforms with previously published data from other countries. PMID- 12529949 TI - [Positional plagiocephaly]. AB - INTRODUCTION: To prevent early sudden infant death syndrome, the health authorities recommend that newborn infants sleep on their back. This has led to an increase of positional plagiocephaly. The aim of this study was to describe this condition and to recommend a preventive treatment. MATERIAL AND METHODS: This is a retrospective registration of 133 children with positional plagiocephaly seen in the period from 1994 to 2000. RESULTS: The number of children with positional plagiocephaly increased from two in 1994 to a maximum of 43 in 1999. 83 were males and 50 were females. 84 were dextral and 49 were sinistral. In seven (14%) of the sinistral and three (4%) of the dextral, we found a physical explanation of the head turning. In 51 children, X-rays of the skull were performed, but no synostosis was found. DISCUSSION: The back-sleeping position of infants is a promotive factor to positional plagiocephaly which may be prevented by simple alternating head positioning. Otherwise early (< 6 months of age) corrective physiotherapy and positioning or an orthoplastic helmet must be considered. The diagnosis of positional plagiocephaly is based on clinical observations. There is a favourite head turning, an ipsilateral occipital flattening, an ipsilateral frontal bossing, and the ipsilateral ear is moved forward. PMID- 12529950 TI - [Mirizzi's syndrome--diagnosed preoperatively by MR scanning]. AB - A case of Mirizzi's syndrome in a 30-year-old woman diagnosed preoperatively by MR-scanning is presented. The MR-scanning showed an impacted stone in the gallbladder neck and surrounding soft-tissue mass compressing the hepatic duct causing a narrow stricture of the duct. There were moderate dilatation of the intrahepatic bile ducts as well as gallbladder stones but the common duct was normal. The findings were consistent with Mirizzi's syndrome, which was confirmed by surgery. Awareness of Mirizzi's syndrome because of its potential complications and MR-scanning as a potential diagnostic tool is advocated. PMID- 12529951 TI - [Spondylodiscitis as a complication to ultrasound-guided transrectal prostatic biopsy]. AB - We report a case of spondylodiscitis and psoas abscess induced by transrectal ultrasound-guided prostatic biopsy in a 70-year-old man. Multi-resistant E. coli was cultured, and the patient was cured after several trials with antibiotic treatments. PMID- 12529952 TI - [Picture of the month: recurrent cervix cancer, hydronephrosis, affected renal function, bladder hematoma]. PMID- 12529953 TI - [Mass spectrometric proteomic analysis of serum samples--a promising method for early diagnosis of cancer]. PMID- 12529954 TI - [Thrombolysis in apoplexy]. PMID- 12529955 TI - [Sex discrimination]. PMID- 12529956 TI - Numerical description of selected endo- and ectocranial dimensions in Homo sapiens and the three Homo heidelbergensis: Kabwe, Atapuerca and Petralona. AB - Investigation of hominid cranial morphology is of particular importance for those dealing with questions concerning both ontogenetic and phylogenetic aspects in the genus Homo. The present study provides a numerical description of several regions of extant human crania and, in addition, mid-Pleistocene crania, given in distances and angles. This study is seen as a basic preparatory work that is most useful for further investigation on cranial development in hominids. The sample used consisted of 60 recent adult human crania as well as stereolithographic models of three H. heidelbergensis crania: Kabwe, Atapuerca and Petralona. 12 ecto- and 20 endocranial landmarks were selected and 3D-coordinates taken on each cranium using a 3D Polhemus 3Space FASTRAK tracking system. From the resulting data set, 21 ectocranial and 17 endocranial distances were calculated, in addition to 41 angles of the ectocranium and 21 angles of the endocranium; the measurements are presented in tables, serving as a reference database. PMID- 12529957 TI - [The skull of Combe Capelle]. AB - Since the end of World War II two of the most important anthropological artefacts of the Museum fur Vor- und Fruhgeschichte in Berlin, the skulls and skeletons of Le Moustier and Combe Capelle, were believed to be missing or destroyed, respectively. The postcrania were severely damaged during a fire after the museum was bombed in February 1945, while the skulls were brought to the Soviet Union in 1945. In 1965, the skull of the Neanderthal man from Le Moustier and the chain of the grave of Combe Capelle were found amongst the art objects returned by the Soviet Union into the German Democratic Republic in 1958. However, the Combe Capelle skull was still missing. In the end of 2001 this skull could be found and identified in a store-house of the museum. Now, one the oldest known representatives of Homo sapiens sapiens is again available for scientific research and public exhibitions. PMID- 12529958 TI - [Goethe's skull and skeleton]. AB - In 1970 there was a conservation of Goethe's skeleton, housed in the Furstengruft in Weimar. This procedure was kept secret until 1999. The restoration of the sarcophagus and the conservation of the skeleton have been documented together with many photos in the so-called "Sonderakte Mazeration Goethe" (special file mazeration Goethe). In this special file there are also photos of Goethe's skull and skeleton which will be published here for the first time and analysed under anthropological and palaeopathological aspects. A short description of the skull will be given. The dentition of Goethe at the time of his death and the question of progenia in Goethe's family are discussed in detail. Data on Goethe's stature obtained from the skeleton qualify statements of Goethe's contemporaries. Among the few documented pathological changes the ankylosis of eight thoracal vertebrae (T 5-12) and five right ribs are of special interest, caused by the loss of the intervertebral discs, spondylosis deformans and Morbus Forestier. A connection between this ankylosis and the known stiff posture and stiff gait of Goethe is obviously and possibly dates back to Goethe's fortieth years of age. PMID- 12529959 TI - A possibility of systematisation of anthropometric data of girls aged 12-15. AB - The article describes an anthropometric cross-sectional study of 374 healthy schoolgirls aged 12-15 years from secondary schools of Tartu (Estonia). 29 body measurements and 9 skinfolds were measured. Mean skinfolds and 6 indices and ratios (including body mass index) were calculated. In each age group the data were systematized into 5 height-weight SD-classes according to the correspondence between body height and weight. The medium class lies between -0.5 SD and +0.5 SD of the respective age group mean (M +/- 0.5 SD); the other classes contain the respective outer values. All the subjects were assigned into one of the following five categories: three height/weight-concordant categories: I = small (small height-small weight), II = medium (medium height-medium weight), III = large (big height-big weight) and two height/weight-disconcordant categories: IV = so-called pyknomorphous, V = so-called leptomorphous. Categories IV and V thus contained three height/weight subclasses each. The body build categories differ significantly from each other within both groups (I-III and IV-V) in all age groups in all measured variables, except sternum length, abdomen length, trunk length, biacromial breadth, biiliocrystal breadth, femur breadth, ankle breadth, humerus breadth, wrist breadth, and head circumference for which no significant differences between pyknomorphous and leptomorphous categories were found. A multidimensional statistical model of body build might be used in girls going through puberty to systematize and compare morphological variables with other assessed characteristics in different applied studies, for example physiological, sociological, psychological, or nutritional. PMID- 12529960 TI - Detection of the p110 beta subunit of phosphatidylinositol 3-kinase complexed with neutral endopeptidase. AB - BACKGROUND: Neutral endopeptidase 24.11 (NEP) is a cell-surface peptidase that inactivates a variety of neuropeptide substrates. In addition to catalytic activity, NEP can exert biological effects through protein-protein interactions. We previously reported that NEP directly associated with tyrosine-phosphorylated Lyn kinase, and with the p85 subunit of the phosphatidylinositol 3-kinase (PI3 kinase) resulting in an NEP-Lyn-PI3 kinase protein complex. MATERIALS AND METHODS: In this report, we investigated the association of NEP with cytoplasmic proteins using ProteinChip Array, surface enhanced laser desorption/ionization (SELDI) technology combined with time-of-flight mass spectrometry, as well as immunoprecipitation and Western blottings. RESULTS: Using immunocapture on the ProteinChip surface, we identified a 122 kDa protein which associates with NEP derived from LNCaP cell lysates which had the identical molecular weight as the beta-subunit of p110 subunit of phosphatidylinositol 3-kinase. The identity of the p110 beta was confirmed by Western blot analysis of NEP and p110 beta immunoprecipitates using monoclonal antibodies specific for NEP and p110 beta. CONCLUSION: These data confirm the association of phosphatidylinositol 3-kinase (consisting of the p85 adaptor and p110 beta-subunit) with NEP. Furthermore, this work demonstrates the ability of mass spectrometry to identify proteins interacting with NEP and potentially other cell-surface peptidases. PMID- 12529961 TI - Specific and non-specific KGF inhibition of KGF-induced breast cancer cell motility. AB - BACKGROUND: Keratinocyte growth factor (KGF), a member of the fibroblast growth factor family, is a mesenchymally derived mediator of epithelial cell proliferation and migration. In a previous study, we reported that KGF enhanced the motility of human breast cancer cells. The objective of the present study was to examine the influence of specific and non-specific KGF inhibitors on KGF induced motility and proliferation in ER-positive MCF-7 cells. MATERIALS AND METHODS: In the present study three KGF inhibitors were employed [Heparin, Innohep, a low molecular weight heparin (LMWH) and KGFR2 beta (IIIb)/Fc, a chimeric KGFR fragment]. Heparin and LMWH bind to low affinity sites on KGF and produce non-specific inhibition, while KGFR2 beta (IIIb)/Fc, a soluble chimera of an extracellular KGFR fragment, is a more specific KGF inhibitor. Cellular motility was measured using two methods: culture wounding over a period of 48 hours; and secondly, time-lapse videomicroscopy (TLVM). RESULTS: In these experiments KGF was found to produce a dose-dependent enhancement of MCF-7 cell motility over a dosage range of 5 to 500 ng/ml. In the TLVM experiments, Heparin (30 ng/ml), LMWH (30 ng/ml) and KGFR2 beta (IIIb)/Fc (50 micrograms/ml) completely inhibited KGF-induced motility of MCF-7 cells during the initial 2 hour observation period. In the culture wounding assay, LMWH produced a greater reduction in KGF-induced motility than heparin at 48 hours post-treatment. CONCLUSION: The results of this study indicate that KGF-mediated enhancement of breast cancer cells motility and proliferation is inhibited by both specific and non-specific KGF inhibitors. LMWH appears to produce an inhibition of KGF with a much longer duration of action than Heparin. Our results suggest that KGF inhibition may be a potential new therapeutic approach for the treatment of metastatic breast cancer. PMID- 12529963 TI - Overexpression of the neutral amino acid transporter ASCT2 in human colorectal adenocarcinoma. AB - Cancer cells have increased amino acid transport and retention. Our aim was to determine whether the neutral amino acid transporter ASCT2 is expressed in colorectal adenocarcinomas (CRC), and whether its expression is of biological significance. Western blot analysis was performed on 4 cases of CRC. Immunohistochemical staining was performed on sections of 63 CRC, and the percent of positive cells was determined. Survival analysis was performed using the Kaplan-Meier method. Western blot analysis showed a 50 kDa protein, corresponding to ASCT2, in 3 of the 4 CRC samples. Immunohistostaining showed lack of ASCT2 immunoreactivity in 41% of the cases. ASCT2 immunoreactivity was present in 1-25% of the cancer cells in 24% of the cases, 26-50% of the cells in 13%, and in > 50% of the cells in 22% of the cases. Patient survival decreased with increased percentage of ASCT2-positive cancer cells (p = 0.0002). These results indicate that ASCT2 is expressed in a significant number of CRC, and that ASCT2 expression is associated with aggressive biological behavior. Larger studies with multivariate analysis are needed to determine whether ASCT2 expression is an independent prognostic marker in CRC. PMID- 12529962 TI - In vivo gene transfer of murine interleukin-4 inhibits colon-26-mediated cancer cachexia in mice. AB - Cancer cachexia has been suggested to be mediated by various cytokines derived either from tumor or host tissue. In the murine colon-26 (C-26) adenocarcinoma model IL-1, secreted by tumor-infiltrating mononuclear phagocytes, has an important role in induction of cancer cachexia. In order to suppress production of IL-1 in peritoneal macrophages we have used liposome-mediated gene transfer of the anti-inflammatory cytokine mIL-4, known as a potent inhibitor of IL-1 production. Balb/c mice were transfected by intraperitoneal inoculation of C-26 tumor cells. The mIL-4 transfected animals showed increased survival rate, delayed symptoms of cachexia and reduced anorexia in comparison with tumor bearing control groups. However, tumor growth inhibition was not seen in mIL-4 transfected animals. Peritoneal macrophages from surviving mIL-4-transfected mice, when stimulated with LPS ex vivo, showed decreased IL-1 alpha production, 1672 +/- 202 pg/2 x 10(6) cells in contrast to tumor-bearing control animals, 3975 +/- 89 pg/2 x 10(6) cells, mock-transfected tumor-bearing animals 4004 +/- 174 pg/2 x 10(6) cells and tumor-free animals, 3142 +/- 60 pg/2 x 10(6) cells (p < 0.004). The present study demonstrates that in vivo gene transfer of an anti inflammatory cytokine reduces cancer-associated cachexia by inhibition of IL-1 alpha production of tumor surrounding peritoneal macrophages, without a significant effect on tumor growth. PMID- 12529964 TI - rIL-18 triggered gene therapy based on a transduction with the IL-12 plasmid: a new option as immuno-therapy for osteosarcoma? AB - Gene therapy is a promising new method to treat tumors locally. Immuno-therapy for treatment of osteosarcomas is one option for hopefully improving the survival rate of patients with this tumor. Transduction of OS cells with the pCMV-IL-12neo plasmid induced a significant increase in IFN-gamma expression by mononuclear cells. This is known to induce antitumor effects mediated by the immune system. In combination with an administration of rIL-18, the IFN-gamma increase was multiplied in a dose-dependent manner. This study demonstrated that osteosarcoma cells can be targeted effectively in vitro by plasmids encoding the IL-12 gene. Considering the synergistic pathways it is reasonable to combine a local, gene transfer based on IL-12 with a rIL-18 administration to trigger the potentially promising immuno-effects for adjuvant treatment of osteosarcomas. PMID- 12529965 TI - Canine mammary tumor cells transfected with B7-1 or B7-2 stimulate proliferation of peripheral blood mononuclear cells. AB - Neoplastic cells are believed to evade the immune system due, in part, to their inability to successfully provide a secondary, costimulatory signal for a T lymphocyte proliferative response. This report describes the generation and investigation of genetically engineered canine mammary tumor (CMT) cells that express canine B7-1, canine B7-2, or human B7-2. These transfected cells were used as stimulators in an allogeneic, costimulation assay. CMT cells transfected with canine B7-1 induced the greatest proliferation (7-fold increase), followed by CMT cells transfected with canine B7-2 (5-fold increase). The specificity of the canine B7-2 stimulatory response was demonstrated by a 38% reduction in proliferation caused by an anti-canine B7-2 blocking antibody. These results suggest that canine mammary tumor cells transfected with canine B7-1 or canine B7 2 may be useful for immunotherapeutic purposes. PMID- 12529966 TI - Basic HGF-like peptides inhibit generation of liver metastases in murine and human tumor models. AB - PURPOSE: We have postulated that the peptide domain(s) of the heparin-binding cytokine(s) might have biological activity, which theoretically could be exploited for modulation of the biological behavior of cancer cells. MATERIALS AND METHODS: We used HGF as a model heparin-binding cytokine and synthesized two HGF beta-chain domains, HHRGK (HGP1) and RYRNKH (HGP2), as well as four variants. As target cells, we used three cancer cell lines (HT25 human colonic, HT168-M1/9 human melanoma and 3LL-HH murine lung carcinoma) all characterized by strong liver metastatic potentials. The effects of peptides on cell proliferation, tumor growth and liver metastasis were evaluated. RESULTS: All the basic penta- or hexapeptides exhibited similar antiproliferative effects in vitro in a dose range of 100-1000 ng/ml. Meanwhile, none of the HGP peptides exhibited significant antitumoral effects on the primary spleen tumors in the form of systemic treatment. However, systemic treatment with HGP1, but not with HGP2, applied at the early phase of the dissemination process, showed an inhibitory effect on liver metastatization of all the tumor lines studied. Furthermore, one out of the four hexapeptides, BP4 (KRKRKR), had similar activity. CONCLUSION: Recent data on the antiangiogenic effects of these basic peptides partially explain the in vivo antimetastatic activity. We suggest the small basic penta-hexapeptides as a new class of biological response modifiers which can modulate the metastatic process. PMID- 12529967 TI - Phenoxodiol (2H-1-benzopyran-7-0,1,3-(4-hydroxyphenyl)), a novel isoflavone derivative, inhibits DNA topoisomerase II by stabilizing the cleavable complex. AB - Cancer therapeutic drugs that inhibit DNA topoisomerase (topo) II by stabilizing the cleavable complex are collectively known as topo II poisons. Phenoxodiol is a synthetic derivative of the plant isoflavone daidzein and is currently undergoing clinical testing as a cancer therapeutic drug. The development of this agent as an antitumor drug was based to a large extent on its low toxicity in normal tissues but potent topo II inhibitory effects in rapidly dividing tumor cells. To evaluate phenoxodiol as a potential inhibitor of topoisomerases, we used the relaxation and nicking assays that can identify topo I inhibitors, and the unknotting and DNA cleavage assays that can identify topo II inhibitors. Phenoxodiol inhibited the catalytic activity of topo II in a dose-dependent manner and it stabilized the topo II-mediated cleavable complex, demonstrating that this agent is a topo II poison. Phenoxodiol's topo II inhibitory effects were comparable to those of other antitumor agents such as VP-16 and were stronger than those of genistein. Phenoxodiol did not inhibit topo I catalytic activity nor did it stabilize the topo I-mediated cleavable complex. These results demonstrate that phenoxodiol is a topo II-specific poison and suggest that this novel agent may find applications in cancer chemotherapy. PMID- 12529968 TI - Antitumor activity of palmitic acid found as a selective cytotoxic substance in a marine red alga. AB - In a previous report, we discussed an extract from a marine red alga, Amphiroa zonata, which shows selective cytotoxic activity to human leukemic cells, but no cytotoxicity to normal human dermal fibroblast (HDF) cells in vitro. In this study, we identified palmitic acid, a selective cytotoxic substance from the marine algal extract, and investigated its biological activities. At concentrations ranging from 12.5 to 50 micrograms/ml, palmitic acid shows selective cytotoxicity to human leukemic cells, but no cytotoxicity to normal HDF cells. Furthermore, palmitic acid induces apoptosis in the human leukemic cell line MOLT-4 at 50 micrograms/ml. Palmitic acid also shows in vivo antitumor activity in mice. One molecular target of palmitic acid in tumor cells is DNA topoisomerase I, however, interestingly, it does not affect DNA topoisomerase II, suggesting that palmitic acid may be a lead compound of anticancer drugs. PMID- 12529969 TI - Loss of Msh2 is not associated with FHIT deletion in breast carcinomas. AB - The FHIT gene is a candidate tumor suppressor gene that has been implicated in the development and progression of breast carcinoma. Recent studies have suggested that Fhit inactivation can be a consequence of defects in mismatch repair proteins, particularly Msh2. Fifty-three breast carcinomas were evaluated for Fhit and Msh2 expression by immunohistochemical staining. The same breast carcinomas were examined for allelic loss at three loci within or near FHIT by PCR-based microsatellite analysis. Seventeen of the 53 breast cancer cases were positive for Fhit protein, and 10 of the 43 informative cases showed FHIT loci deletion. Twenty-five of the 53 (47%) cases showed loss of Msh2 expression. No relationship between Msh2 protein loss and FHIT locus alteration or Fhit protein loss could be observed. Our results suggest that this mismatch repair protein may play little role in maintaining the integrity of the common fragile locus with the FHIT gene. PMID- 12529970 TI - Novel system for generating cytotoxic effector lymphocytes using carcinoembryonic antigen (CEA) peptide and cultured dendritic cells. AB - We have established a practical system for generating antitumor effector lymphocytes using the tumor antigen peptide CEA and cultured dendritic cells (DCs), and have also characterized effector cells. DCs were induced from the adherent cell population of autologous peripheral blood mononuclear cells (PBMCs) obtained from HLA-A0201 normal or tumor-bearing donors using IL-4 and GM-CSF. The cultured DCs were shown to express class I, class II, CD80 and CD86 molecules. The PBMCs were stimulated for 7 days with the DCs pulsed with the HLA-A0201 restricted CEA peptide CEA9 671 and then expanded in an anti-CD3 antibody (1 microgram/ml)-coated flask in the presence of a 80 U/ml IL-2 (IL-2/CD3 system). The effector cells, which were designated as CEA peptide-pulsed dendritic cell activated killer (CEA-PDAK) cells, were preferentially CD3+CD8+, and capable of killing T2 cells pulsed with CEA peptide but not T2 cells alone. The CA-PDAK cells also lysed the gastric cancer cell line KATO III (HLA-A0201, CEA (+)), but not the WiDr (HLA-A2402, CEA(+)) cells. The cytotoxicity was abrogated when the CEA-PDAK cells were treated with anti-TCR alpha beta antibody or when the target cells were treated with the anti-class I antibody prior to the cytotoxicity assay. The CEA-PDAK cells exerted their cytotoxic activity even in the presence of a high amount of CEA protein at the effector phase, which mimicked the clinical setting. The CEA-PDAK cells showed approximately a hundred-fold expansion in total cell numbers yielded without any loss of the specific lysis, when stimulated with the IL-2/CD3 system compared to those stimulated with IL-2 alone. The TCR V beta gene analysis for the CEA-PDAK cells, conducted by means of RT-PCR-Southern blotting, demonstrated oligoclonal expression of TCR beta 7 and 12, and the latter was shown to be responsible for the killing activity. SSCP analysis indicated the clonotype of the TCR V beta 12 gene, indicating a selective expansion of lymphocytes bearing a limited TCR variable region by the stimulation with CEA peptide-pulsed DCs. Taken together, the effector lymphocytes reactive with the CEA antigen can be generated from PBMCs with the antigenic CEA peptide and cultured DCs. The IL-2/CD3 system is effective and practical in activating the effector cells for the clinical use of CEA-PDAK cells. Adoptive immunotherapy using this system may be promising for treating CEA-expressing tumors. PMID- 12529971 TI - IFN gamma-induced up-regulation of PD-ECGF/TP enhances the cytotoxicity of 5 fluorouracil and 5'-deoxy-5-fluorouridine in bladder cancer cells. AB - BACKGROUND: PD-ECGF/TP is an essential enzyme in converting 5'DFUR and 5FU to their active metabolites and can be up-regulated by some cytokines. MATERIALS AND METHODS: PD-ECGF/TP mRNA and protein expressions were determined by RT-PCR and Western blot, respectively. The cytotoxicity of 5FU, 5'DFUR or MMC against RT-4 and T24 cells was evaluated by MTS assay. The PD-ECGF/TP expressions in primary bladder cancers were also analyzed. RESULTS: Levels of PD-ECGF/TP mRNA and protein were concomitantly elevated in RT-4 and T24 cells after IFN gamma treatment. IFN gamma decreased the IC50 of 5FU and 5'DFUR in both cell lines, while it did not alter the IC50 of MMC, which is not a substrate of PD-ECGF/TP. PD-ECGF/TP expression correlated with tumor stage and grade in primary bladder cancers. CONCLUSION: IFN gamma enhances the cytotoxicity of 5FU and 5'DFUR against human bladder cancer cells through induction of PD-ECGF/TP. The results imply that an IFN gamma/5FU or IFN gamma/5'DFUR combination therapy may be applicable to clinical bladder cancers. PMID- 12529972 TI - Mechanism of beta 2-microglobulin-induced apoptosis in the K562 leukemia cell line, defective in major histocompatibility class 1. AB - BACKGROUND: It has been shown that exogenous beta 2-microglobulin (beta 2m) triggers significant apoptosis in several cell lines, but the molecular mechanism of beta 2m-induced apoptosis remains to be found. MATERIALS AND METHODS: To understand the mechanism of beta 2m-induced apoptosis, we added purified human beta 2m to cultures of K562 human chronic myelocytic leukemia cells, detected apoptosis by DNA fragmentation and annexin V binding assays, measured mitochondrial membrane potential (delta psi m), and used Z-VAD-fmk, a general inhibitor of caspases, inhibitors of caspases-1 and-3, as well as Western blot analysis to detect activated caspases. RESULTS: beta 2m-induced apoptosis was associated with decreased delta psi m K562 cells. Treatment with the general caspase inhibitor Z-VAD-fmk, as well as the caspase-1 inhibitor YVAD-CHO, significantly blocked beta 2m-induced apoptosis. However, Western blot analysis revealed that caspase-1 was intrinsically activated in untreated as well as beta 2m-treated K562 cells. Furthermore, beta 2m-induced apoptosis was not associated with the cleavage of caspase-3 as revealed by Western blot analysis, but was inhibited by an inhibitor of caspase-3, Z-DEVD-CHO, suggesting that not caspase 3, but a caspase-3-like enzyme may be involved in beta 2m-induced apoptosis. Western blot analysis also revealed that caspases-4, -8 and -9 were not activated during beta 2m-induced apoptosis in these cells. CONCLUSION: These results reveal that beta 2m-induced apoptosis in K562 cells occurs by a mechanism dependent on decreased mitochondrial delta psi m. Moreover, while caspase-1 activity may be one of the factors involved in beta 2m-induced apoptosis, activation of this caspase alone does not cause apoptosis, and other proapoptotic factors including activation of a caspase-3-like enzyme, independently of caspases-4, -8 and -9 activity, may be required to trigger apoptosis. PMID- 12529973 TI - Effects of long-term administration of cancer-promoting substances on oral subepithelial mast cells in the rat. AB - The role of oral subepithelial mast cells in the defence against tumours is a matter of controversy. The effect of established and suggested carcinogens, such as the carcinogen 4-nitroquinoline-N-oxide (4-NQO) and Herpes simplex virus type 1 (HSV-1), in combination with oral snuff on lower lip subepithelial mast cells (MC) was studied in rats. The rats were exposed to prolonged use of oral snuff. The test substances were administered in a surgically created canal in the lower lip of the rats. There were 15 rats in each test group and 10 rats in the control group. The amount of countable subepithelial mast cells decreased significantly when the rat oral mucosa was exposed to the oral carcinogen 4-NQO but the effect of oral snuff and HSV-1 infection was weak. Our findings suggest that mast cells play a role in immunological cell defence against chemical carcinogens. Further studies are needed to clarify the mechanisms. PMID- 12529974 TI - Reduced expression of coxsackievirus and adenovirus receptor (CAR) in tumor tissue compared to normal epithelium in head and neck squamous cell carcinoma patients. AB - Even though adenoviral vector is widely used in gene therapy of squamous cell carcinoma of the head and neck (SCCHN), the expression level of Coxsackievirus and adenovirus receptor (CAR) in SCCNH is not clearly defined. To identify this variability, the expression of CAR was measured using SCCHN cell lines and compared with transfection efficiency. It was found by RT-PCR and Western blot analysis that CAR levels varied in SCCHN cell lines. FACS analysis and adenovirus infection assay revealed that there was a good correlation between the level of CAR expression and the transfection efficiency. To identify the actual CAR expression patterns of human SCCHN tissues in vivo, immunohistochemical staining was undertaken on frozen biopsies of six SCCHN patients. In all the patients examined, the normal tissues showed much stronger staining for CAR than the tumor tissues. These results demonstrate that the level of CAR expression of a tumor should be evaluated before clinical application of adenoviral vector for gene therapy in SCCHN. PMID- 12529975 TI - Glut-1 glucose transporter expression in esophageal squamous cell carcinoma is associated with tumor aggressiveness. AB - BACKGROUND: Glucose transporter (Glut) proteins, which are membrane proteins responsible for the transport of glucose across cellular membranes, have six forms. To further elucidate the role of Glut-1 expression in esophageal squamous cell carcinoma, we examined the expression of Glut-1 protein immunohistochemically. MATERIALS AND METHODS: Immunohistochemical expression of Glut-1 was examined in surgically resected tissues from 95 patients with esophageal squamous cell carcinoma. RESULTS: Of the 95 esophageal carcinomas, 91 (95.8%) had some Glut-1 immunostaining in the membranes of the cancer cells. Positive staining (> 30% of cancer cells showing Glut-1 expression) was observed in 49 (51.6%) of the cases. Comparison of Glut-1 expression and clinicopathological characteristics in the 95 patients with esophageal cancer revealed significant associations between Glut-1 expression and tumor status (p < 0.001), lymph node status (p < 0.05), metastatic status (p < 0.01), and pathological stage (p < 0.001). The survival rates of patients with Glut-1 positive tumors were significantly lower than those of patients with Glut-1 negative tumors (log-rank p < 0.05). CONCLUSION: In conclusion, the level of Glut 1 expression may be a useful marker that can provide information on tumor aggressiveness and prognosis in patients with esophageal squamous cell carcinoma. PMID- 12529976 TI - Differential endothelial CAM-expression after stimulation with supernatants of LPS- and cytokine-stimulated HT-29 and ST-ML-12 tumor cells growing as monolayer cultures and multicellular spheroids. AB - The mechanisms of tumor metastasis in vitro have been principally investigated using monolayer culture systems. In vivo, however, multicellular clusters of tumor cells rather than individual cells appear to penetrate the microvasculature. Thus, the multicellular tumor spheroid model represents a more suitable tool to study tumor cell-endothelial interactions. Our interest has centered on the role of inflammation on tumor intra- and extravasation. The expression of three endothelial cell adhesion molecules (CAM), ICAM-1, VCAM-1 and E-selectin, on HUVEC after incubation with supernatants of previously LPS-, TNF alpha- or IL-1 beta-stimulated HT-29 (colon adenocarcinoma) and ST-ML-12 (melanoma) tumor cells growing as monolayers and spheroids, was compared using cell enzyme immunoassay. The results indicated that important differences in the expression of CAM between monolayers and spheroids exist depending on the tumor cell line. The possible significance of LPS for colon carcinomas was underlined by the spheroid-model results. With respect to the influence of intestinal bacteria on the behaviour of colon carcinomas, the spheroid model could be a useful in vitro system for a more realistic simulation of in vivo conditions. PMID- 12529977 TI - Are critical normal gene products in cancer cells the real therapeutic targets? AB - The majority of targets for rational anti-cancer drug development available at present have been defined by studies of early carcinogenesis. In this situation, mutations resulting in altered expression or structure of gene products, in the context of a relatively normal diploid background, provide the cell with a growth advantage. In contrast to cells studied in early carcinogenesis, however, profound chromosomal abnormalities and widespread disruption of gene expression can be found in the cells of typical cancers seen in the clinic. In addition tumour heterogeneity and genetic instability, in clinical cancers, can result in variable and elusive drug targets. The severe degree of genetic damage, seen in the clinical situation, would be expected to sufficiently disrupt the function of cell-signalling pathways, such that the highly complex and co-ordinated process of cell division, which occurs in normal cells, could no longer be effectively accomplished in the clinical cancer cell. It may therefore be relevant to consider whether certain Critical Normal Genes must be retained throughout the life of the progressively evolving cancer cell in order to enable it to continue to divide successfully. The regulatory roles of the products of such Critical Normal Genes would be expected to become particularly important in the chaotic molecular environment, which finally occurs in the typical clinical cancer cell. Here remaining Critical Normal Gene Products may be required to perform novel functions to maintain cellular integrity. Because the function of such gene products would be essential for continued cancer cell proliferation, they would not be expected to be subject to tumour heterogeneity or genetic instability as loss of function of a gene product essential to cell proliferation would inevitably result in the death of that cell. Perhaps such remaining Critical Normal Gene Products, with adapted functions critical to continued cancer cell proliferation, whose disruption may only have minimal effects within the multiple signalling pathways of normal cells and which are likely to be unaffected by tumour heterogeneity and genetic instability, could provide particularly relevant targets for anti-cancer drug development. PMID- 12529978 TI - Evaluation of carcinogenic potential of two nitro-musk derivatives, musk xylene and musk tibetene in a host-mediated in vivo/in vitro assay system. AB - We have developed a host-mediated assay system for detection of the transforming activity of chemical carcinogens on peritoneal macrophages. Directly, as well as indirectly acting carcinogenic substances, administered intraperitoneally to NMRI mice, could be examined in this way. Resident macrophages were recovered by peritoneal lavage from treated and untreated mice and cultured in soft agar. After 5-6 days the normal and transformed cells could be distinguished. Statistical analysis comparing cells from musk xylene- or musk tibetene-treated animals with those from control mice proved that the test is positive. Musk xylene and musk tibetene revealed a cell-transforming potential that showed a dose-dependent response in our host-mediated assay system. We have succeeded in establishing permanent cell lines from mice treated with musk xylene, or musk tibetene. The oncogenicity of these cell lines was tested in athymic nu/nu mice. Animals injected subcutaneously with these cells (1 x 10(6) cells at each side of the neck) developed tumors at the injection sites within 3 weeks of treatment. The experimental data reported here lead to the conclusion that musk xylene, as well as musk tibetene, have carcinogenic activity. In contrast to the negative results for mutagenicity and genotoxicity, a non-genotoxic mechanism for the carcinogenicity of musk xylene and musk tibetene must be considered. PMID- 12529979 TI - Tumor sensitivity to anti-cancer drugs predicted by changes in fluorescence intensity and polarization in vitro. AB - BACKGROUND: We used the CellScan, a novel static cytometer, to monitor changes induced by anti-neoplastic drugs in the fluorescence intensity and polarization of fluorescently-labeled tumor cells. MATERIALS AND METHODS: T47D and T80 human breast cancer cell lines were exposed to navelbine and to 5-fluorouracil and the fluorescence properties of the treated cells, stained with fluorescein diacetate and rhodamine 123, were measured by the CellScan. RESULTS: A strong correlation was found between the inhibition of cell growth induced by the two drugs, as estimated from cell counts, and the resulting changes in fluorescence intensity and polarization, as monitored by the CellScan. Fluorescence hyperpolarization of the labeled cells occurred in conjunction with AnnexinV binding and propidium iodide exclusion, indicating that such hyperpolarization, resulting from drug action, reflects an early stage of apoptosis, as previously proposed. CONCLUSION: The system presented here could serve as the basis for assessing drug sensitivity or resistance of cancer cells derived from small biopsies of solid human tumors, thus eliminating prior tumor culturing and time-consuming assays. PMID- 12529980 TI - Role and prognostic significance of CD44s expression in colorectal cancer. AB - The purpose of this study was to clarify the role and the predictive strength of the adhesion molecule CD44s (standard isoform) in colorectal carcinogenesis. MATERIALS AND METHODS: CD44s immunohistochemical expression was evaluated in 100 patients with colon adenoma and 100 patients with colon adenocarcinoma and adjacent non-neoplastic mucosa (ANNM). The patients were followed-up for five years. RESULTS: CD44s immunoreactivity was expressed in low-moderate-high-grade dysplasia adenomas and associated with adenocarcinoma (p = 0.01), ANNM (p = 0.05) and pTNM stage (p = 0.00001). Univariate analysis revealed that CD44s expression was associated with overall survival (OS) in carcinomas (p = 0.01) and ANNM (p = 0.05). Bivariate analysis revealed that CD44s was associated with OS in stages I and II patients (p = 0.03). Multivariate analysis revealed that stage (p = 0.0001) and CD44s expression (p = 0.05) were independent predictors of OS. CONCLUSION: CD44s is involved in colon carcinogenesis and is associated with aggressive carcinomas. The immunohistochemical expression of CD44s may reveal cells that have lost their adhesion ability and therefore detect carcinomas with high metastatic power. PMID- 12529981 TI - Cytotoxic effects of cuphiin D1 on the growth of human cervical carcinoma and normal cells. AB - Cuphiin D1 (CD1), macrocyclic hydrolyzable tannin isolated from Cuphea hyssopifolia, has been shown to exert an antitumor effect both in vitro and in vivo. Furthermore, CD1 significantly inhibited the growth of the human cervical carcinoma, i.e. HeLa, cells and showed less cytotoxicity to normal primary cultured cervical fibroblasts. In this study, we explored the cytotoxic mechanism of CD1 on HeLa cells. The cytotoxic effects of CD1 showed dose-dependency at 3.15 100 micrograms/ml on HeLa for 12, 24, 48 and 72 hours and with an IC50 value at 14.2 micrograms/ml for 48 hours. However, the IC50 value of CD1 in primary cultured normal cervical fibroblasts was 74.5 micrograms/ml. Therefore, the selectivity shown by CD1 is ascribed to differences in growth speeds between normal and tumor cells. HeLa cells treated with 50 micrograms/ml CD1 for 24 hours exhibited chromatin condensation, indicating the occurrence of apoptosis. Flow cytometric analysis demonstrated the presence of apoptotic cells with low DNA content among HeLa cells. CD1 also caused DNA fragmentation and inhibited Bcl-2, pro-caspase 3, and inactived PARP expression in HeLa cells. These results suggest that the inhibition of Bcl-2 expression in HeLa cells might account for the mechanism of CD1-induced apoptosis. PMID- 12529982 TI - Induction of apoptosis by iron depletion in the human breast cancer MCF-7 cell line and the 13762NF rat mammary adenocarcinoma in vivo. AB - It is known that the interruption of normal iron metabolism with chelators of iron, toxic metals, toxic metals bound to transferrin, or anti-transferrin receptor antibodies leads to significant inhibition of tumor cell growth in cell culture systems and animal models. In the present study, we found that iron depletion was produced by the iron chelator deferoxamine mesylate, the free toxic metals gallium or indium, and the toxic metals gallium or indium bound to transferrin in the MCF-7 human breast cancer cell line, and this induced the condensation and fragmentation of chromatin, and the formation of DNA fragments characteristic of apoptosis. The induction of apoptosis was quantitated with acridine orange and ethidium bromide staining of apoptotic cells, separation of fragmented DNA from radiolabeled cells, and in situ terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) assays. The apoptosis, caused by deferoxamine mesylate, and gallium or indium bound to transferrin in the MCF-7 cells, can be completely inhibited by excess ferric chloride or equimolar iron-loaded transferrin. Gallium-transferrin and indium transferrin complexes induced more apoptosis than their respective salts in the MCF-7 cells. Deferoxamine mesylate induced a small increase in the endogenous expression of both the bcl-2 and bax genes in the MCF-7 cells and this can be prevented by ferric chloride. In the 13762NF rat mammary adenocarcinoma model, in situ TUNEL assays showed that the iron-deficiency following a low iron diet or intravenous injection of deferoxamine mesylate produced 5.32 +/- 3.90% and 6.46 +/- 3.58% of apoptotic cells, respectively, compared to 2.01 +/- 1.20% of apoptotic cells in the control rats maintained on a normal diet (p < 0.05 and p < 0.01, respectively, Student's t-test). This is the first report of iron depletion caused by a low iron diet or deferoxamine mesylate treatment inducing apoptosis in rats bearing the 13762NF marnmary adenocarcinoma. PMID- 12529983 TI - Characterization of a fragment containing a putative TLP cDNA sequence. AB - With the aim of isolating the Tumor Liberated Protein (TLP) gene, reverse transcription-polymerase chain reaction (RT-PCR) was used to isolate a approximately equal to 500 bp fragment containing a putative TLP cDNA sequence. Total RNAs were extracted from several cell lines with RNAzol B reagent (Tel Test, Inc) and reverse transcribed using the Reverse Transcription System (Promega). PCR was carried out for 35 cycles (1 minute at 95 degrees C, 2 minutes at 40 degrees C and 1 minute at 72 degrees C) using an upstream degenerate oligonucleotide, ACN AAY AAR GAR GCN TCN ATG TG, which corresponds to the amino acid sequence T N K E A S I, and random hexamers as the downstream primer. PCR products were electrophoresed on a 1% agarose gel containing ethidium bromide. The PCR products were cloned in the pGEM-T easy vector (PROMEGA) and the resulting plasmid clones were sequenced with the chain termination method using the Applied Biosystems model 373A DNA sequencer. A putative open reading frame was deducted. The results obtained can be considered as preliminary data that will require more investigation in order to confirm them. We propose to continue the studies to verify that TLP could be a diagnostic marker in human cancer. PMID- 12529984 TI - Hypoxia-induced changes in the expression of VEGF, HIF-1 alpha and cell cycle related molecules in ovarian cancer cells. AB - BACKGROUND: Hypoxia is important in cancer progression, and at the stage of detachment of the cancer cells from the primary lesion. This study was undertaken to analyze the effect of hypoxia on angiogenesis and cell proliferation in ovarian cancer. MATERIALS AND METHODS: We first used immunohistochemistry to examine the expression of VEGF in 42 cases of ovarian carcinoma, with relevance to the p53 expression. Then, the expression of VEGF, HIF-1 alpha, cell cycle related molecules and cell numbers were examined in 4 ovarian cancer cell lines with various p53 gene status. RESULTS: Immunohistochemistry showed that there was no significant correlation between VEGF and p53 expression. Moreover, hypoxia increased the expression of VEGF via up-regulation of HIF-1 alpha irrespective of p53 gene status. However hypoxia did not change the cell numbers, but influenced the expression of cell cycle-related molecules (increased p27 and decreased cyclin D1 and Rb). CONCLUSION: Hypoxia increased VEGF expression in ovarian cancer cells irrespective of p53 gene status. PMID- 12529985 TI - Evidence for a direct antitumor mechanism of action of bovine lactoferricin. AB - BACKGROUND: Bovine lactoferrin (LFB) and its pepsin-generated peptide lactoferricin (LfcinB) possess antitumor activities. The mechanism underlying the antitumor activities of LfcinB in vivo has not been elucidated. In this study the antitumor activities exerted by LFB, LfcinB and murine lactoferricin (LfcinM) on murine tumor cell lines and experimental tumors were investigated. MATEIALS AND METHODS: The protein and peptides were tested against Meth A fibrosarcoma, B16F10 melanoma and C26 colon carcinoma cells in vitro and their derived tumors in vivo, exploring the mechanisms of antitumor activity by way of histological and scanning electron microscopical studies. RESULTS: LfcinB exerted significant cytotoxic activity against the three tumor cell lines in vitro and significantly reduced the size of solid Meth A tumors. Scanning electron micrographs revealed tumor cell membrane disruption and eventually cell lysis, while extensive hemorrhagic necrosis was apparent in tumor sections one day after LfcinB treatment. No species-specific antitumor effect of LfcinM was observed. CONCLUSION: Our study demonstrated that LfcinB elicits an antitumor effect mediated through a direct mechanism of action not observed with LFB or LfcinM. PMID- 12529986 TI - Radical scavenging activity and cytotoxicity of ferulic acid. AB - Ferulic acid and eugenol were examined for their superoxide (O2-), hydroxyl radical (.OH) and nitric oxide (NO)-scavenging ability, using ESR spectroscopy with spin trap agents DMPO and carboxy-PTIO/NOC-7. Ferulic acid more efficiently scavenged .OH and NO than eugenol. The O2- scavenging activity of ferulic acid was comparable with that of eugenol. Ferulic acid significantly reduced the NO production by lipopolysaccharide (LPS)-stimulated mouse macrophage-like cells (Raw 264.7 cells) compared to eugenol. The cytotoxic activity of ferulic acid against Raw 264.7 cells was comparable with that against human submandibular gland carcinoma (HSG) cells and the cytotoxicity of ferulic acid was about 10 fold smaller than that of eugenol. The stoichiometric factor (n) (number of moles of peroxy radical trapped by moles of the relevant phenol) of ferulic acid and eugenol was investigated, using the induction period methods of the methyl methacrylate polymerization system. The n-value of ferulic acid (1.5) was higher than that of eugenol (1.0) and was similar to that of 2, 6-di-t-butyl-4 methylphenol (BHT). Ferulic acid as well as eugenol may produce a dimer during the induction period due to an n-value less than 2. These results suggested that ferulic acid may be useful for preventing cell damage perhaps caused by O2-, and in particular by .OH and NO, in living systems. PMID- 12529987 TI - Effect of lignins and their precursors on nitric oxide, citrulline and asparagine production by mouse macrophage-like Raw 264.7 cells. AB - Lignins, tannins and flavonoids are commonly found polyphenols. Among these polyphenols, lignins, polymers of phenylpropenoids complexed with polysaccharides, were the least cytotoxic and most potently stimulated the production of nitric oxide (NO), citrulline and asparagine by mouse macrophage like Raw 264.7 cells. The maximum production of these substances reached the level attained by lipopolysaccharide (LPS). However, epigallocatechin gallate, phenylpropenoid monomers (ferulic acid, caffeic acid) and gallic acid (component unit of tannin) were inactive. These data suggest that the macrophage-stimulation activity of polyphenols depends, at least in part, on their molecular weight or structural configuration. There was a positive relationship between the extent of asparagine production and that of NO or citrulline. Western blot analysis demonstrated that both lignins and LPS elevated the cellular level of asparagine synthetase. The present study suggests the possible link between the stimulated asparagine production and macrophage activation. PMID- 12529988 TI - Prostate-specific antigen (PSA)-mediated proliferation, androgenic regulation and inhibitory effects of LY312340 in HOS-TE85 (TE85) human osteosarcoma cells. AB - BACKGROUND: PSA mediates growth factor responses that stimulate proliferation of prostatic and other cellular types. Androgen-sensitive TE85 human osteosarcoma cells were used to study PSA as a potential mediator of prostatic cancer growth and osseous metastasis. MATERIALS AND METHODS: TE85 cells were probed for PSA mRNA and protein levels under testosterone (T)-replete and--depleted conditions. TE85 proliferative responses to PSA were evaluated in the absence and presence of LY312340, a monocyclic beta-lactam inhibitor of PSA enzymatic activity. RESULTS: A 3.1-fold increase in PSA mRNA was observed following T stimulation. Low levels of immunoreactive PSA (iPSA) were detected in media of androgen-stimulated TE85 cells while iPSA was not found in control media. Conversely, iPSA was detected in TE85 cell pellets from control but not in androgen-stimulated cell cultures. Exogenously added enzymatically active PSA stimulated TE85 proliferation in a bi phasic manner. LY312340 inhibited PSA-induced increases in TE85 cell numbers but had no effect on basal or T- stimulated cellular proliferation. CONCLUSION: While the PSA levels produced by TE-85 cells in response to androgen stimulation are too low to be biologically active, PSA produced by metastatic PCa cells may mediate paracrine stimulation of osteogenic PCa metastasis. Inhibitors of PSA enzymatic activity could be useful therapeutic agents. PMID- 12529989 TI - Oral mucosal carcinogenesis in SENCAR mice. AB - Animal models of oral carcinogenesis have been developed but most use the hamster buccal pouch or rat oral mucosa. With completion of human and murine genome sequencing, the development of a mouse model of oral carcinogenesis may prove useful for future genomic studies of oral carcinogenesis. To achieve this objective, 30 SENCAR mice were initiated by brush application of palatal, buccal and tongue mucosa with 200 nmol 7,12-dimethylbenz[a]anthracene (DMBA) using 3 treatment regimens, and promoted by brush application with 5 nmol 12-O tetradecanoylphorbol-13-acetate (TPA) for a total of 28 weeks. Alternatively, 5 mice were treated with 0.5% 4-nitroquinoline-1-oxide (4NQO) alone by brush application for 28 weeks. There were another 6 control mice treated with vehicle alone. The tumor samples were analyzed for the presence of H-ras codon 61 gene mutations using a mutant-allele-specific amplification-polymerase chain reaction (MASA-PCR) technique. The results showed that among the group of 24 mice initiated with DMBA for 2 or 6 weeks, a range of papilliferous lesions were seen on the buccal mucosa comprising papillomas, papillomas with dysplasia and 7 squamous cell carcinomas (SCC). In those 6 mice initiated with 1 week of DMBA, only papillomas developed. In the 5 mice treated with 4NQO, one developed papillomas with dysplasia and two had SCCs in the tongue mucosa but not the buccal mucosa. Both carcinogens induced codon 61 mutation of the H-ras gene at a high frequency. The results indicated that DMBA/TPA and 4NQO in SENCAR mice reliably produced preneoplastic and malignant oral cavity lesions, which resemble the multistages for human oral carcinogenesis, and targeted to site-specific zones of the oral mucosa, namely the buccal mucosa and tongue, respectively. These results show that SENCAR mice can be used as a unique model of oral carcinogenesis with the potential for detailed molecular studies of neoplastic progression to SCC. PMID- 12529990 TI - Signal transduction pathway to low-dose radiation-induced apoptosis in peripheral PNET cells. AB - We investigated the signal transduction pathway to low-dose radiation-induced apoptosis in vitro in the human peripheral primitive neuroectodermal tumor (pPNET) cell line with wild-type p53 established in our laboratory. Apoptosis was induced by 2Gy irradiation in an almost p53-dependent manner in this model except for a deficiency of the cleavage of caspase-9. It was detected 3 hours after irradiation by fragmentation assay. The expressions of p53, p21WAF-1 and Bax increased, in contrast to the gradually decreasing expression of Bcl-2, as observed by immunoblotting. Following this, cleavages of caspase-3 and PARP reached peak levels. There were no detectable increases in ERK expression and caspase-9 cleavage. In respect of the probability of other pathways to apoptosis, this cell line will provide a useful model both for investigating low-dose radiation-induced signal transduction pathway and for analyzing the biological characteristics of pPNET. PMID- 12529991 TI - Correlative polymorphism of NAD(P)H: quinone oxidoreductase (NQO1) with telomere shortening in colorectal cancer. AB - AIM: The aim of this study was to examine whether and relationships could be found among polymorphism of the NQO1 gene, telomere length and telomerase activity in colorectal cancers. MATERIALS AND METHODS: Fifty-one invasive colorectal cancers were studied. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was undergone to detect mutation of the NQO1 gene. Telomere length was examined by Southern blot analysis. Telomerase activity was assayed by telomeric repeat amplification protocol with minor modifications. RESULTS: Of the 51 tumors, 20 (39.2%) and 9 (17.6%) were heterozygous and homozygous for the mutation, respectively. Most of the cases homozygous for the mutation (88.9%) showed short telomeres and its frequency was significantly higher than in those heterozygous (p = 0.0432). However no relationship was found between the telomerase activity and mutation in the NQO1 gene. CONCLUSION: Our data suggest that oxidative stress by the lack of NQO1 activity could result in telomere shortening through colorectal cancinogenesis. PMID- 12529992 TI - 5-aza-2'-deoxycytidine induces retinoic acid receptor beta 2 demethylation, cell cycle arrest and growth inhibition in breast carcinoma cells. AB - Like all cancers, breast cancer is considered to result in part from the accumulation of multiple genetic alterations leading to oncogene overexpression and tumor suppressor loss. More recently, CpG island hypermethylation is known to be associated with gene silencing in cancer, and these silenced genes can be reactivated by 5-aza-2'-deoxycytidine (5-Aza-CdR). Retionoic acid receptor beta 2 gene is a tumor suppressor gene and the chemopreventive effects of retinoids are due to induction of RAR beta 2. In this study, the effect of 5-Aza-CdR RAR beta 2 restoration was investigated in the MRK-nu-1 human female breast cancer cell line. Changes of the RAR beta 2 methylation status were assessed by methylation specific PCR. Reverse transcription PCR was used to evaluate RARb beta 2 restoration. Cell cycling and growth inhibition were studied using flow cytometric analysis of DNA content and CellTiter 96 AQueous non-radioactive cell proliferation assay, respectively. 5-Aza-CdR treatment resulted in complete demethylation of the RAR beta 2 gene. RAR beta 2 restoration was accompanied by cell cycle arrest (increase in the G0/G1- and decrease in the S- and G2/M-phases) and time-dependent growth inhibition. In conclusion, RAR beta 2 can be activated in vitro by 5-Aza-CdR, which may be one of the mechanisms for the tumor cell growth inhibition by 5-Aza-CdR. PMID- 12529993 TI - Synthetic retinoid CD437 induces cell-dependent cycle arrest by differential regulation of cell cycle associated proteins. AB - BACKGROUND: The synthetic retinoid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2 naphthalene (CD437) exhibits a wide spectrum antitumor activity through induction of cellular apoptosis and cell cycle arrest. We investigated the effects and mechanisms of CD437 on cell cycle arrest of gastric cancer cells. MATERIALS AND METHODS: The activities of CD437 on cell growth were analyzed by measuring total cellular DNA. The effects of CD437 on cell cycle phase distribution were analyzed using flow cytometry. The levels of cell cycle associated proteins were analyzed by Western blot. The activities of cyclin-dependent kinases (cdks) were analyzed by phosphorylation of the histone H1 protein. RESULTS: CD437 at concentrations between 0.1 and 10 microM profoundly suppressed the growth of all six gastric cancer cell lines. Growth suppression associated with induction of G0/G1 or G2/M arrest was cell-line-dependent. CD437 decreased levels of cdk inhibitor p21 in G2/M-arrested SC-M1 cells. However, CD437 increased p21 levels in G0/G1-arrested AGS cells. Total and activated cyclin-dependent kinases were differentially regulated by CD437 in AGS and SC-M1 cells. CD437 (1 microM) induced activity of cdk2- and p34cdc2-associated H1 kinase by 14.6- and 1.8-fold, respectively, in SC M1 cells. In contrast, CD437 slightly increased (1.6-fold) the cdk2-associated H1 kinase activity in AGS cells. CONCLUSION: CD437 profoundly suppressed the growth of gastric cancer cells, which was associated with cell-dependent induction of G0/G1 or G2/M arrest. The differential regulation of p21 that leads to alteration in the activity of cdks may play a critical role in cell-line-dependent regulation of cell cycle arrest following treatment with CD437. PMID- 12529994 TI - Lobaplatin as an adjuvant chemotherapy to surgery in canine appendicular osteosarcoma: a phase II evaluation. AB - Canine osteosarcoma, the most common bone tumor in dogs, is a well-established, naturally-occurring animal model for human OS. The aim of this study was to evaluate the clinical and hematological side-effects and to assess the efficacy of lobaplatin chemotherapy in dogs with appendicular osteosarcoma as an adjuvant therapy to surgical resection. Twenty-eight dogs without systemic signs of disease were treated with surgical resection of the tumor and adjuvant lobaplatin chemotherapy at a dose of 35 mg/m2, i.v., once every three weeks, for a maximum of 4 doses. Clinical signs of toxicosis were uncommon and consisted mainly of vomiting and depression. Hematological signs of toxicoses were common 7 to 10 days after lobaplatin chemotherapy and consisted of thrombocytopenia, leukopenia and neutropenia. All the signs were transient and most disappeared within three weeks of lobaplatin administration. A one-year disease-free fraction of 21.8% and a one-year survival fraction of 31.8% were calculated. Multivariate Cox regression analyses showed that a high histological tumor grade and presence of metastasis in the tumor vessels were associated with significantly shorter disease-free interval and survival time. Also, an increased pretreatment plasma alkaline phosphatase level at first presentation and a high histological level of tumor necrosis were associated with a shorter survival interval. Lobaplatin was easy to administer as an i.v. bolus injection at a three-week interval in dogs without the need for pretreatment infusions. PMID- 12529995 TI - Tumour cell uptake of lactosaminated and intact N-succinyl-chitosans and antitumour effects of conjugates with mitomycin C. AB - BACKGROUND: The relationship between the uptake of lactosaminated and intact N succinyl-chitosans into hepatoma cells (MH134 and AH130) and liver metastatic tumour model cells (M5076) and the antitumour effects of their conjugates with mitomycin C (MMC) were investigated. MATERIALS AND METHODS: Fluorescently labelled carriers were administered to tumour-bearing mice. The fluorescence intensity and microscopic examinations were performed at 1 hour post-injection. The antitumour effects were examined according to several schedules: one of them was the administration of each conjugate at a dose of 4 mg eq. MMC/kg x 4 days at 3 days post-inoculation. RESULTS: A difference in uptake was found by measurement of fluorescence intensity between both carriers only in MH134 cells, but was not recognized by fluorescence microscopy. Among these cell lines, the uptake of carriers into M5076 cells tended to be the most extensive. The difference in antitumour effects of the conjugates against MH134 and M5076 was reflected by the biodistribution study. CONCLUSION: The pattern of antitumour effects was markedly different among cell lines of different origins. PMID- 12529997 TI - Cytotoxicity of paclitaxel in combination with cisplatin and a new Pt mercaptopyridine complex. AB - In vitro synergy has been reported for exposure to paclitaxel prior to cis-platin (cis-DDP), whereas the reverse sequence resulted in antagonism. There is no clear evidence for an intracellular origin of the schedule-dependent interaction, but several hypotheses have been proposed, such as effects at the level of DNA crosslinks, or binding to tubulin sites. The purpose of this study was to evaluate the cytotoxicity of these two drugs as single agents, in combination and in sequence, against a human colon adenocarcinoma cell line (LoVo). Moreover, we considered the new Pt-mercaptopyridine complex C/2, was like cis-platin in being able to alter DNA conformation. We have therefore studied the cytotoxic effects after single agent exposure, concomitant exposure (paclitaxel + cis-platin and paclitaxel + C/2) and sequential drugs exposure (paclitaxel-->cis-platin and cis platin-->paclitaxel or paclitaxel-->C/2 and C/2-->paclitaxel). Our results demonstrate that the most cytotoxic effect is induced by paclitaxel and C/2 exposure both in the case of concomitant cell treatment and sequential exposure paclitaxel-->C/2. PMID- 12529996 TI - Partial purification of cytotoxic substances from moxa extract. AB - The major cytotoxic activity of Moxa was extracted with CH2Cl2 and partially purified by three cycles of silica gel column chromatography. The active fractions showed higher cytotoxicity against six human tumor cell lines (two oral squamous cell carcinoma, one salivary gland tumor, one melanoma, two leukemia) than three normal oral human cells (gingival fibroblast, periodontal ligament fibroblast, pulp cell). All fractions failed to protect the cells from the cytopathic effect induced by HIV infection. ESR spectroscopy showed that all fractions produced little or no radical under alkaline conditions, while showing much lower O2- scavenging activity, generated by hypoxanthine-xanthine oxidase reaction, than antioxidants and polyphenols. Active fractions induced DNA fragmentation in HL-60 cells, but failed to modify the mobility and activity of mitochondrial Mn-containing superoxide dismutase (MnSOD), in contrast to Moxa smoke. These data suggest that the active principles in the Moxa extract might be different from that in Moxa smoke, which produced carbon radical and modified MnSOD mobility and activity. PMID- 12529998 TI - Immunophenotypic changes and induction of apoptosis of monocytes and tumour cells during their interactions in vitro. AB - The in vitro model of tumour infiltrating macrophages (TIM)-tumour interactions in which monocytes and monocyte-derived macrophages (MDM) are cultured with cancer cells was used to assess immunophenotypic changes of interacting cells. Following short cocultures, monocytes, MDM and tumour cells were sorted out by FACS and the expression of several determinants was evaluated. Monocytes showed the induction of CD44v6 and v7/8, and up-regulation of CD16 (Fc gamma RIII), CD54 (ICAM-1), CD68 (macrophage maturation marker) and CD86 (costimulatory molecule B7.2). The increased expression of CD11a (LFA-1) and CD58 (LFA-3) was noted on some cancer cells. Up-regulation of TNFRII and HLA-DR was observed on both types of cells. MDM shared similar changes. Contact of monocytes, but not of MDM, with tumour cells led to Fas-FasL-dependent apoptosis of both types of cells. This study suggests that the immunophenotype of monocytes/macrophages and cancer cells may be modified during their bidirectional interactions in the absence of other microenvironmental elements that are present in the tumour stroma. PMID- 12529999 TI - Effect of S. cerevisiae APN1 protein on mammalian DNA base excision repair. AB - Mammalian cells transfected with the S. cerevisiae APN1 protein acquire resistance to oxidizing agents, the damage of which are mainly repaired via DNA base excision repair (BER). We have recently hypothesized that this effect might be linked to the possible capacity of APN1 to accelerate mammalian BER by its 3' diesterase activity. We have investigated here the effect of pure APN1 protein on BER performed by mouse embryonic fibroblast extracts. No significant acceleration was observed in the repair of either a single AP site cleaved by the bifunctional glycosylase NTH of E. coli or the repair of a single 8-oxoguanine, initiated by the bifunctional glycosylase OGG1. Similarly, no significant effect was observed on the repair of a single U (initiated by the monofunctional glycosylase U DNA glycosylase) or the repair of a single natural abasic site. The inability of APN1 to increase the efficiency of BER initiated by bifunctional glycosylases indicates that removal of 3' blocking fragments is not the rate limiting step of this repair pathway. PMID- 12530000 TI - Association of thymidylate synthase gene polymorphism with its mRNA and protein expression and with prognosis in gastric cancer. AB - Thymidylate synthase (TS) is a target enzyme of 5-fluorouracil (5-FU). TS has a polymorphic repeated sequence in the 5'-UTR and the polymorphism is associated with TS protein expression. TS polymorphism has been reported to link with the efficacy of 5-FU-based chemotherapy in colorectal cancer. In this study, we examined whether the association among TS polymorphism, TS mRNA expression and TS protein expression is also observed in gastric cancer tissues and whether the TS polymorphism is a prognostic factor for patients with gastric cancer. We quantified TS mRNA isolated from 115 gastric cancer tissues by real-time reverse transcription PCR and TS protein in 72 available samples by the fluoro-dUMP binding assay. These values were compared with TS genotypes of the samples determined by a PCR assay. The tumor with the 3R/3R genotype had higher TS protein expression than that with the 2R/3R genotype, although there is no association between the TS genotype and mRNA expression. The result suggests that mRNA translation is responsible for the genotype-dependent difference of TS protein expression, as is consistent with our previous observation in colorectal cancer. This association encouraged us to further examine the link of the TS genotype with the stage of disease and the prognosis of the patients. The clinicopathological analysis showed that gastric cancers with the 3R/3R genotype are at a more advanced stage than those with the combined 2R/2R and 2R/3R genotype. We observed a longer survival in those patients with the combined 2R/2R and 2R/3R genotype, compared with the 3R/3R genotype, although it did not reach significance when the patients who had received the oral fluoropyrimidines therapy were analyzed. These results warrant further large-scale clinical study of the role of the TS genotyping for the prediction of efficacy using 5-FU-based chemotherapy and prognosis in gastric cancer. PMID- 12530001 TI - The combined effects of antibacterial peptide cecropin A and anti-cancer agents on leukemia cells. AB - Anti-microbial cecropins are humoral immune peptides originally found in insects. They possess a particular function of membrane permeabilization on both gram positive and gram-negative bacteria. Yet, they are not capable of lysing eucaryotic cells. In this experiment, we confirmed that cecropin A (CA) is a special agent able to lyse leukemia cells while exerting little toxic effect on normal lymphocytes, based on the IC50 measurements. The therapeutic index of CA ranged from 6 to 17. In contrast, chemical anti-cancer agents (ACAs) are non specific drugs capable of damaging both cancer and normal cells. Due to the functional differences between CA and ACAs, the experiments on the combined effects of CA and ACAs were conducted in vitro. Under certain conditions, synergistic effects were found with CA/S-FU and CA/Ara-C combinations. The results suggest the need for further study into the possible application of combining anti-cancer agents with antibacterial peptides for cancer treatment. PMID- 12530002 TI - Anti-prostate cancer and anti-breast cancer activities of two peptides derived from alpha-fetoprotein. AB - A chemically synthesized 34-amino-acid peptide and a select analog have been studied to determine their activities against the growth of prostate and breast cancer tumors. It was of interest to determine if the peptide has anti-prostate cancer activity. Previously, the peptide was shown to inhibit the growth of breast cancer tumor cells. The peptide inhibited the growth of both breast and prostate tumors. A novel experimental design for the peptide was in a study in which a time-release pellet was used to give daily peptide doses to mice that were subjected to a breast cancer tumor. The peptide was effective in inhibiting the growth of tumors in the mice. The 2 C-->2 A analog peptide, in which the two cysteines were replaced by alanines, was also active in inhibition of the growth of prostate and breast cancer cell lines and in an in vivo assay against breast cancer. A scrambled amino acid sequence of the peptide was used as a control in these tumor studies, and it had virtually no anti-cancer activity. PMID- 12530003 TI - Amifostine protects blood vessels from the effects of ionizing radiation. AB - Amifostine (WR-2721) is a well-known radioprotective drug, selective for normal cells. The purpose of the present study was to define whether amifostine protects the vascular network from the effects of X-rays. We used the in vivo system of chicken embryo chorioallantoic membrane (CAM) as a model of angiogenesis. Amifostine reversed the early X-rays- induced decrease in the number of CAM blood vessels and reversed the early radiation-induced apoptosis of CAM cells. It also inhibited the increase in tyrosine nitration of actin and a-tubulin, which was observed 6 hours after CAM irradiation, when there was a significant decrease in non-protein SH groups. Furthermore, C6 rat glioma cells were inoculated on CAM and tumor growth, as well as tumor-induced angiogenesis, was estimated on haematoxylin-eosin-stained paraffin sections. Amifostine inhibited the post irradiation increase of C6 tumor-induced angiogenesis. These data suggest that amifostine protects CAM cells and blood vessels from the effects of X-rays, through mechanisms that do not depend solely on its free radical scavenging properties. PMID- 12530004 TI - Mechanism of cytotoxicity of turbinamide in vitro. AB - BACKGROUND: Previously, we demonstrated that turbinamide isolated from the marine ascidia Sydnium Turbinatum, exerts a selective cytotoxicity on glioma cells. Now we have investigated the mechanism of its cytotoxic effect on two different cell lines: C6 rat glioma cells and J774 murine monocyte/macrophages. MATERIALS AND METHODS: Cell viability, membrane lipoperoxidation, DNA fragmentation and apoptosis were studied in C6 and J774 cells incubated with turbinamide for 24 hours. RESULTS: Turbinamide (0.01-100 micrograms/ml) induced a dose-dependent inhibition of C6 cell viability (88.0 +/- 3.0%; 80.0 +/- 1.2%; 38.0 +/- 5.0%; 23.6 +/- 1.3% and 9.1 +/- 2.3%) with respect to the control (100% viability). Moreover, turbinamide (0.01-100 micrograms/ml) increased lipoperoxidation of C6 cells (1.21 +/- 0.11; 9.03 +/- 1.6; 17.8 +/- 1.6 and 45.03 +/- 1.01 ngMDA/1 x 10(6) cells) with respect to unstimulated cells (0.6 +/- 0.1 ngMDA/1 x 10(6) cells) that was accompanied by DNA damage, having no effect on J774. Interestingly, turbinamide (0.1-100 micrograms/ml) induced apoptosis in C6 cells. CONCLUSIONS: Our study showed that the selective cytotoxicity of turbinamide to C6 cells is due to apoptosis, suggesting that turbinamide may be useful in the therapy of glioma. PMID- 12530005 TI - Interaction between various resistance modifiers and apoptosis inducer 12H benzo[alpha]phenothiazine. AB - The effect of some resistance modifiers on apoptosis induction by a benzo[alpha]phenothiazine derivative was studied on the L5178Y mouse lymphoma cells (parent) and its multidrug resistant (MDR) subline. For evaluation of apoptosis the cells were stained with FITC-labelled annexin V and propidium iodide and the results were analysed by flow cytometry. 12H benzo[alpha]phenothiazine [M627] induced apoptosis both in the parent cells and in the MDR cells. The apoptosis induction by [M627] was not affected significantly by post- or pre-treatment with resistance modifiers, while in the cells treated by (+/-)-verapamil before and after apoptosis induction with [M627], the apoptosis was somewhat higher. The resistance modifier compounds alone also induced apoptosis and it was slightly higher in the parent cells than its MDR1/A gene-transformed subline. PMID- 12530006 TI - Activation of the anti-apoptotic Akt/protein kinase B pathway in human malignant gliomas in vivo. AB - In the present study we investigated two important signal transduction pathways in glioma biopsies. By Western analysis we found an overexpression of the epidermal growth factor receptor in 10 out of 27 (37%) glioblastoma multiforme (GBM), but not in astrocytomas WHO II/III which demonstrated only weak or absent expression. Only two GBM (8%) but none of the astrocytomas WHO II/III exhibited loss of PTEN expression. Activation of Akt/protein kinase B showed a close correlation with EGF receptor overexpression in human malignant gliomas since 6 out of 7 GBMs with high degrees of protein kinase B activation exhibited overexpression of the EGF receptor. In contrast, no significant differences in MAP kinase activation could be detected between individual GBMs. Our data show that EGF receptor overexpression seems to be responsible for activation of the protein kinase B whereas PTEN deletion seems to play a minor role in the dysregulation of this important pathway in human GBM in vivo. PMID- 12530007 TI - In vitro effects of genistein on the synthesis and distribution of glycosaminoglycans/proteoglycans by estrogen receptor-positive and -negative human breast cancer epithelial cells. AB - Genistein, a soy isoflavone, affects the proliferation of both estrogen receptor (ER)-positive and ER-negative cancer cells. Glycosaminoglycans (GAGs)/proteoglycans (PGs) are considered to be of great importance in the treatment of cancer. The synthesis of GAGs by two human breast cancer epithelial cell lines, the ER-positive MCF-7 and the ER-negative BT-20, was studied under the effects of genistein, and their distribution in the culture medium and the cell membranes was determined. The results obtained show that both cell lines synthesize extracellular hyaluronic acid (HA) and both extracellular and cell associated galactosaminoglycans (GalAGs) and heparan sulphate (HS). The MCF-7 cell line synthesizes HA, GalAGs and HS at considerably lower rates than the BT 20 cell line. The effect of genistein on the synthesis of extracellularly secreted GAGs/PGs by ER-positive MCF-7 cells is dose-dependent and follows two mechanisms; one at low concentrations (< or = 35 microM) mediated via the estrogen receptor and the other at higher concentrations via protein tyrosine kinase (PTK). The synthesis of cell-associated GAGs/PGs by ER-positive MCF-7 cells and of both secreted and associated with the cell membrane GAGs/PGs by ER negative BT-20 cells is mediated by a PTK mechanism. It is concluded that genistein affects the synthesis of GAGs/PGs, by breast cancer epithelial cells depending on the presence or absence of estrogen receptor and the localisation of PGs. PMID- 12530008 TI - Endogenous tetrapeptide from neuroblastoma and new-born pig brain inhibits neuroblastoma cell growth in vitro. AB - Neuroblastoma, the most frequent malignant tumour in neonates and young children, has an unusual clinical behaviour, age being the most important single factor. This could indicate that some growth-regulating mechanism in lost, or changed, during the first years of life. In search for possible growth-modifying factors, we identified a tetrapeptide, acetyl-Asp-Gln-Tyr-GlyNH2, in extracts of neuroblastoma tissue, in neuroblastoma cell cultures and in new-born pig brain tissue. The purified native peptide as well as a synthetic peptide with the same structure decreases in vitro growth of neuroblastoma cells at a restricted, low (picomolar) range. The structure of the peptide as well as its dose-response characteristics, indicates that it belongs to a group of endogenous growth modifying oligopeptides that previously have been isolated from other organs and tissues. A possible role for the new peptide in clinical medicine is discussed. PMID- 12530009 TI - Cytostatic and apoptosis-inducing activity of boswellic acids toward malignant cell lines in vitro. AB - Boswellic acids from frankincense were indentified as the active compounds which inhibit leukotriene biosynthesis, 5-lipoxygenase and exert antiproliferative activity toward a variety of malignant cells. Because of the relevance for the clinical application, we tested the ethanolic extract of Boswellia serrata gum resin containing a defined amount of boswellic acids for its cytotoxic, cytostatic and apoptotic activity on five leukemia (HL-60, K 562, U937, MOLT-4, THP-1) and two brain tumor (LN-18, LN-229) cell lines by WST-1 assay and flow cytometry. The Boswellia serrata extract induced dose-dependent antiproliferative effects on all human malignant cells tested with GI50 values (extract concentration producing 50% cell growth inhibition) between 57.0 and 124.1 micrograms/ml. In three haematological cell lines (K562, U937, MOLT-4) the effect of total extract expressed in GI50 was 2.8-, 3.3- and 2.3-times more potent (p < 0.05) than pure 3-O-acetyl-11-keto-beta-boswellic acid (AKBA). Morphological changes after 24-27 hours and the detection of apoptotic cells by AnnexinV binding and/or by the detection of propidium iodide-labelled DNA with flow cytometry, confirmed the apoptotic cell death. The results of this study suggest the effectiveness of Boswellia serrata extract with defined content of boswellic acids. PMID- 12530010 TI - Inhibition of P-glycoprotein transport function by N-acylphenothiazines. AB - Multidrug resistance (mdr) constitutes the major obstacle to successful cancer treatment. The ability of fifteen newly-synthesised N-acylphenothiazine derivatives to inhibit the transport activity of P-glycoprotein was studied by flow cytometry in a resistant mouse lymphoma cell line. A standard functional assay with rhodamine 123 as a fluorescent substrate analogue was used. All derivatives proved to be effective inhibitors of rhodamine 123 outward transport, however their toxicity to the cells was not negligible. Phenothiazine maleates probably interact with transporter proteins of cancer cells by a different mechanism than other phenothiazine derivatives studied. The mdr reversal mechanism of phenothiazine acetylamides, methoxycarbonylamides and methylsulfonylamides is likely to involve modulator-cell membrane interactions since a connection between the compounds' hydrophobicity and their P-glycoprotein inhibition potency was observed. As a result of the present study a new group of mdr reversal agents was identified. PMID- 12530011 TI - Detection of occult breast cancer cells by amplification of CK19 mRNA by reverse transcriptase-polymerase chain reaction: role of surgical manipulation. AB - BACKGROUND: To assess if surgical manipulation increases peripheral blood cancer cells dissemination in early stage (I and II) breast cancer patients. PATIENTS AND METHODS: We analyzed 64 patients using RT-PCR for cytokeratin-19 as a marker for peripheral blood breast cancer cell dissemination. Peripheral blood was obtained at 4 different time-points (24 hours before and after surgery, one week and one month later). RESULTS: RT-PCR was positive in 14 (24%) out of 59 evaluable patients. Circulating cells were detected in 4 out of 14 patients before surgery (7%) while in the remaining 10, the positivity was observed after surgery (17%). The percentage of patients with occult breast cancer cells increased significantly after surgery (p = 0.01). CONCLUSION: 1) 7% of early breast cancer patients had circulating tumor cells before surgery. 2) After surgery tumor cells were detected in 17% of patients. 3) Surgery significantly increased the presence of occult breast cancer cells. 4) The clinical significance of occult breast cancer cells should be tested within a larger clinical trial trying to assess their role as an independent prognostic factor. PMID- 12530012 TI - Anticancer, antiradical and antioxidative actions of novel Antoksyd S and its major components, baicalin and baicalein. AB - Here we show for the first time that the novel designed drug, Antoksyd S and its polyphenolic flavones (baicalin and baicalein), act as cell proliferation modifiers of mouse leukemia cells (L1210). The cytotoxicity of Antoksyd S and baicalein in vitro was expressed as ED50 and compared with those of the cytostatics doxorubicin, cisplatin and DACA, under the same experimental conditions. Cell viability was determined by modified tetrazolium dye assay, using as a model cells with neoblastic phenotype (L1210). The antiradical activity of Antoksyd S and baicalin were investigated using the DPPH test in order to obtain their antioxidant characteristics. Structure- and concentration dependent one electron bioactivations (peroxidative oxidation) of Antoksyd S and baicalin were performed in the absence or in the presence of GSH or SOD. It appears that Antoksyd S is a low toxic novel drug which could be effective in providing concentration-dependent antioxidative activity, acting as a cell proliferation modifier and, probably, as an apoptosis inducer in vitro, though this remains to be explored. These findings are discussed from a mechanistic standpoint as well as in terms of potential pharmacological applications under acute oxidative stress and apoptotic events. This work provides the basis for further investigations of Antoksyd S action in vitro and in vivo, since the presented results are indicative of its intracellular metabolic activation, which can only be partially associated with its observed action towards model cancer cells (mouse leukemia L1210). PMID- 12530013 TI - Compositional and structural alterations of proteoglycans in human rectum carcinoma with special reference to versican and decorin. AB - This study indicated that human normal rectum (HNR) and human rectum carcinoma (HRC) contained three populations of proteoglycans (PGs). About 63% of the HNR PGs, in terms of uronic acid, were heparan sulfate proteoglycans (HSPGs) of M(r) 500,000 with HS side-chains of M(r) 35,000. The other two populations were versican (29%) and decorin (8%) of M(r) 715,000 and 90,000, respectively, bearing mainly dermatan sulfate (DS) (73%) and chondroitin sulfate (CS) (27%) chains of M(r) 24-26,000 and 20-22,000, respectively. In contrast, in terms of uronic acid, HRC contained 2-fold amounts of PGs. The majority of these PGs (87%) were versican and decorin of lower hydrodynamic size (500,000 and 70,000, respectively) than in HNR, with CS as prominent GAG (70%) in both types of PG. The M(r) of CS and DS chains in these PGs was 12-14,000 and 14-16,000, respectively. The remaining portion (13%) of PG was HSPGs of lower hydrodynamic size (300,000) with smaller HS chains (29,000) than HSPGs of HNR. Moreover, the molar concentrations of versican and decorin estimated from PG-derived protein contents represented a significant, but disproportionate increase, about 5-fold and 8-fold, respectively. The sulfation pattern of rectum carcinoma-associated versican and decorin was significantly altered mainly in containing (62%) 6 sulfated disaccharides and a significant proportion (10%) of non-sulfated disaccharides. DS chains of the tumor-associated versican and decorin contained decreased amounts of iduronic acid. On the metabolic level, the abnormally high production of versican and decorin in this malignant tumor suggests a dramatic modification in their biosynthetic steps at both translational and posttranslational levels. PMID- 12530014 TI - Immunohistochemical detection of C-kit (CD117) and vascular endothelial growth factor (VEGF) overexpression in mantle cell lymphoma. AB - BACKGROUND: Mantle cell lymphoma (MCL) is a low-grade lymphoproliferative malignancy that is extremely refractory to chemotherapy. Commonly used treatments have yielded unfavorable response rates (30% complete remission). We evaluated the incidence of c-kit (CD117) and vascular endothelial growth factor (VEGF) overexpression in patients with MCL in an effort to identify possible targets for therapeutic intervention. MATERIALS AND METHODS: Patients with a diagnosis of MCL based on CD5 positivity associated with cyclin D1 positivity and CD23 negativity on the lymph node/bone marrow specimen were included in our retrospective study. CD117 overexpression was performed using immunohistochemistry on archival specimens. VEGF expression was detected by the avidin-biotin-complex method. RESULTS: Between 1997 and 2001, we identified 17 patients with MCL (9 males, 8 females) with a mean age of 57 years (age range: 42-66 years). The mean overall survival was 34 months (range: 11-60 months). VEGF expression was identified in 7 out of 17 (41.18%) patients with MCL. Among the VEGF-positive patients (n = 7, 41.1%), the mean survival was 24 months (range: 11-42 months), while patients without VEGF expression (n = 10, 58.9%) had a mean survival of 44 months (range: 21-60 months). CD117 expression was identified in only 2 out of 17 (1.17%) patients in our study. CONCLUSION: Our study evaluated the role of c-kit and VEGF overexpression in MCL. Although CD117 may not be of therapeutic significance, target-directed signal transduction inhibition therapy using VEGF-inhibitors may be a distinct possibility in a select group of patients with MCL. Future larger studies are urgently needed to elaborate the role of VEGF in MCL. PMID- 12530015 TI - Red cell Hb oxidation of healthy subjects compared to breast cancer patients. AB - The end result of our research would be the development of a new diagnostic technique that could be used as a screening test, starting from a simple blood collection, a common procedure without any risk to the subject. Red blood cells (RBC) of patients affected by breast cancer are more sensitive to the denaturing action of acetylphenylhydrazine (APH), hence the formation time of hemin, one of the last oxidation products, is significantly shorter then in normal subjects. Our previous experiment showed the differences during iron oxidation between breast cancer patients and healthy persons (Croci et al. 2001). Different amounts of hemin, after the same incubation time, can discriminate between samples collected from people suffering from cancer and a healthy population. These results have given birth to the necessity to standardise a procedure useful as a screening method. The aim of the present work was to define a range of values that characterise the samples collected from healthy people. PMID- 12530016 TI - ATPase inhibitors suppress actinomycin D-induced apoptosis in leukemia cells. AB - BACKGROUND: Apoptosis is mediated by many kinds of enzymes such as caspases, DNase and protein kinases. Recently, ATPase has been shown to be involved in the apoptotic system, but its role is not fully understood. MATERIALS AND METHODS: We investigated the effect of 8 species of ATPase inhibitors on actinomycin D plus colcemid-induced apoptosis in human megakaryoblastic leukemia CMK-7 cells by monitoring caspase-3 activation and DNA cleavage. RESULTS: 2,3-Butanedione monoxime (BDM), lansoprazole, cyclopiazonic acid, geldanamycin and radicicol suppressed the apoptosis. Among these compounds, geldanamycin was the strongest suppressor of both caspase-3 activation and DNA cleavage. Furthermore, Western blotting showed that radicicol suppressed the proteolytic cleavage of procaspase 9 more strongly than BDM, lansoprazole or cyclopiazonic acid. CONCLUSION: Since geldanamycin and radicicol are specific inhibitors of the ATPase in HSP90, the present result implies that ATPase activity in HSP90 plays some role in this apoptosis. PMID- 12530017 TI - Significance of plasminogen-activation system in the formation of macroscopic types and invasion in esophageal carcinoma. AB - BACKGROUND: The roles of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) in the formation of macroscopic types and invasion were investigated. MATERIALS AND METHODS: A total of 40 surgically resected esophageal carcinoma tissues were immunohistochemically stained, and the expression of uPA, PAI-1 and the uPA/PAI-1 ratio, were evaluated. RESULTS: The expression of uPA was significantly stronger in the macroscopically infiltrative type (n = 20: p = 0.0027), whereas PAI-1 was significantly stronger in the localized type (n = 20: p = 0.0005). The uPA/PAI-1 ratio was significantly higher in the infiltrative type (p < 0.0001). A significant correlation was found between the U/P ratio and the depth of tumor invasion (r = 0.511, p = 0.0014). Analysis of tumors of uniform size (4.0-6.0 cm in length), showed that the depth of invasion was significantly greater in the infiltrative type (p = 0.0038). CONCLUSION: The results demonstrated that uPA and PAI-1 play important roles in invasion and formation of macroscopic types of esophageal carcinoma. PMID- 12530018 TI - Bicalutamide dose-dependently inhibits proliferation in human prostatic carcinoma cell lines and primary cultures. AB - OBJECTIVE: Androgen antagonists inhibit prostatic cell proliferation in normal and pathological conditions and are useful antitumor agents in prostatic carcinoma (PCa). Bicalutamide (BCLT) is a well-known non-steroidal antiandrogenic agent able to interfere with androgen receptor (AR). We tested the efficacy of BCLT in inhibiting proliferation of human PCa cell lines and of primary cultures from biopsies of PCa patients. MATERIALS AND METHODS: Human prostatic carcinoma cell lines (PC3, DU145, LNCaP ALVA 31 and ND1) and short-term primary tissue cultures from PCa patients were treated with BCLT. Cell proliferation and orange acridine and ethidium bromide fluorescence staining studies were performed. RESULTS: BCLT was able to inhibit, significantly and dose-dependently, cell proliferation in AR-positive human PCa cell lines and in 10 cases of primary cultures with Gleason grades 4 to 8. Its action appears to be mainly apoptotic in AR-positive cells and cytotoxic in AR-negative cells. CONCLUSION: BCLT, which inhibits growth in both human PCa cell lines and PCa primary cultures from patients with medium and low-grade tumors, deserves attention as a potential widely effective antiandrogenic monotherapy in prostatic carcinoma. However, its efficacy in AR-negative cells requires further research. PMID- 12530019 TI - Treosulfan in the treatment of advanced ovarian cancer: a randomised co-operative multicentre phase III-study. AB - BACKGROUND: In August 1988 a randomised phase III multicenter trial was started in order to compare cisplatinum/treosulfan (PT) with standard cisplatinum/cyclophosphamide (PC) in advanced ovarian carcinoma, aiming at lower toxicity and maintained efficiency. PATIENTS AND METHODS: Five hundred and nineteen patients were enrolled into the protocol. Final evaluation after a median observation time of more than five years was made in July 1996 and included 398 eligible patients, of whom 366 were evaluable regarding efficiency and 290 in respect of toxicity. The tumour stages were classified as FIGO II in 53, FIGO III in 244 and FIGO IV in 68 patients. The patients were stratified regarding post-operative tumour burden. RESULTS: Hematological and gastrointestinal toxicity WHO > = 3 were comparable between the two study arms though a significant difference could be demonstrated regarding alopecia (PT 8% vs. PC 47% after six cycles). The median time to progression as the main efficiency item was in favour of the study schedule (PT 20.6 vs. PC 15.1 months) while significant differences were neither observed in the whole study group nor in the analysed subgroups (R0, < 2 cm, > = 2 cm). The same held true for overall survival. CONCLUSION: PT may be recommended as a less toxic substitute for the former standard PC. After the acceptance of paclitaxel/cisplatin as a new standard, the role of treosulfan should be investigated regarding adjuvant therapy in patients without residual tumor, as a potential partner in triple or sequential treatment and in second-line treatment. PMID- 12530020 TI - LEA.135 expression: identifies low-risk patients with breast ductal carcinoma in situ. AB - The expression of a cell surface-associated sialoglycoprotein (LEA.135), which has been shown to be significantly associated with decreased incidence of recurrence and increased overall survival of patients with primary invasive breast carcinoma, was evaluated in a retrospective study to identify subsets of patients with breast ductal carcinoma in situ (DCIS) who are at high risk of subsequently developing invasive breast carcinoma. The study was carried out by an immuno-histochemical method on formalin-fixed and paraffin-embedded tissue sections from 63 patients initially with DCIS. Pathological parameters such as DCIS histological type, nuclear grade, as well as time and type of recurrence (either a second DCIS or the diagnosis of locally invasive breast carcinoma) together with follow-up in years were available for the cohort of patients. A comparison of recurrence was made of patients whose tumor cells exhibited LEA.135 expression (24 +/- 8% recurring by 7 years), compared with those patients whose specimens showed the absence of LEA.135 expression (41 +/- 10% recurring by 7 years). A statistically significant univariant association between LEA.135 expression and the absence of recurrence of DCIS or development of locally invasive breast carcinoma was observed, suggesting a favorable prognostic significance of LEA. 135 expression (log-rank p = 0.032). It is worthy of mention that none of the LEA. 135-positive patients developed recurrence as DCIS or locally invasive breast carcinoma (0.24 +/- 0.08) after 5 years of the initial diagnosis of DCIS, whereas those from LEA. 135-negative progressively increased their recurrence at 5 years (0.30 +/- 0.09), 7 years (0.41 +/- 0.10) and 10 years (0.63 +/- 0.12). The results of this pilot study show that LEA.135 expression is significantly associated with a favorable prognosis of patients with DCIS, leading to a decreased incidence of recurrence. PMID- 12530021 TI - Tandem high-dose chemotherapy supported by autologous peripheral blood stem cell transplantation for recurrent soft tissue sarcoma. AB - BACKGROUND: Patients with recurrent soft tissue sarcoma (STS) are seldom curable, with 5-year survival rates of less than 10% in all large series. The role of high dose chemotherapy (HDC) with hematopoietic stem cell support in this disease has not been established. CASE REPORT: We report on two patients with recurrent STS who were treated with tandem HDC supported by autologous peripheral blood stem cell transplantation (PBSCT). One patient with malignant fibrous histiocytoma recurred with multiple lung metastases. This patient achieved a partial response after two cycles of induction chemotherapy consisting of ifosfamide and epirubicin. During four cycles of induction chemotherapy, peripheral blood stem cells (PBSCs) were harvested. Tandem high-dose ICE regimen (ifosfamide 3 g/m2 on days-7 to -3, carboplatin 400 mg/m2 on days-7, -5 and 3, etoposide 500 mg/m2 on days-7, -5 and 3) supported by autologous PBSCT gave rise to further regression of the tumors. Another patient with malignant hemangiopericytoma was treated by tandem high-dose ICE regimen supported by autologous PBSCT after the 3rd removal of abdominal tumors. Relapse-free intervals until the 1st, 2nd and 3rd relapses were 40, 19 and 22 months, respectively. Tandem high-dose ICE regimen might delay the relapse. CONCLUSION: These observations suggest that a tandem high-dose ICE regimen with autologous PBSCT is feasible with some clinical efficacy in the control of refractory STS. PMID- 12530022 TI - PTEN expression in tamoxifen-associated endometrial cancers. AB - Tamoxifen is associated with increased rates of endometrial hyperplasia and adenocarcinoma. Our previous work suggested tamoxifen-associated endometrial cancers might be associated with p53 mutations. PTEN, a tumor suppressor gene, is altered in low-grade endometrial carcinoma. This study evaluates PTEN immunohistochemical (IHC) expression in tamoxifen-associated endometrial cancers. MATERIALS AND METHODS: Twenty-eight endometrial carcinoma specimens were examined from patients with a history of breast cancer. Patients who had taken Tamoxifen (15) were compared to non-users (13). IHC staining was performed for PTEN; overexpression was defined as greater than 70% positivity. RESULTS: The mean duration of tamoxifen use was 3.3 years (3-171 months). Four out of 15 (27%) tamoxifen users expressed PTEN compared with 2 out of 13 (15%) of non-users. CONCLUSION: In this study, it appears that tamoxifen-associated endometrial cancers are not significantly different from sporadic endometrial cancer with regards to PTEN IHC expression, although there is a trend towards retained PTEN expression. PMID- 12530023 TI - Tissue polypeptide specific antigen (TPS) as a tumor marker in renal cell carcinoma. AB - Tissue polypeptide specific antigen (TPS) is a new tumor marker that indicates tumor proliferative rate rather than tumor burden. The purpose of this study was to assess the clinical value of TPS in patients with renal cell carcinoma (RCC). Serum levels of TPS were measured in 30 patients with locoregional and in 20 patients with advanced disease before and after therapy. The results showed that: (1) the detection sensitivity of TPS for RCC is 60.0%; (2) the detection sensitivity of TPS in advanced RCC (100.0%) was significantly higher than in locoregional RCC (33.3%); (3) the 10 locoregional RCC patients without recurrence had normal serum TPS levels during a follow-up period of 1 year; and (4) the 8 advanced RCC patients with good response during therapy had normal serum TPS levels, while 12 patients with poor disease had significantly elevated serum TPS levels during a follow-up period of 1 year. Our results suggest that TPS may have a potential clinical role as a valuable tumor marker for RCC, especially in advanced diseases and follow-up therapy response. PMID- 12530024 TI - Biological properties of biopsy specimens are useful for predicting lymph node micrometastasis in esophageal carcinoma. AB - BACKGROUND: Some studies have reported on lymph node micrometastasis (MM) by RT PCR. We attempted to predict MM by biological means using preoperative biopsy specimens. MATERIALS AND METHODS: Lymph nodes from 60 patients with esophageal carcinoma were examined by routine histological examination and CEA-specific RT PCR. The biopsy specimens were immunohistochemically examined using p53, cyclin D1 (CD1) and desmoglein 1 (DG1) antibodies. RESULTS: Of 659 lymph nodes, 53 (8.0%) nodes were positive according to histological examination and 158 (24.0%) had MM by RT-PCR. The percentage of patients with lymph node metastasis according to histological examination and RT-PCR was 65.0% and 81.7%, respectively. CD1 and DG1 expression correlated with MM, whereas p53 expression did not. MM was frequently detected in the tumors with CD1-positive, DG1-negative or reduced expression. CONCLUSION: The expression of CD1 and DG1 in biopsy specimens may offer useful information on lymph node metastasis, including MM in esophageal carcinoma. PMID- 12530025 TI - Correlation of plasma level and immunohistochemical expression of vascular endothelial growth factor in patients with advanced colorectal cancer. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is thought to be the most potent angiogenic factor and may contribute to the progression of various cancers. In colorectal cancer, the immunohistochemical expression of VEGF is reported to be an independent prognostic factor, and elevated plasma levels of VEGF are reported to be a prognostic marker. The purpose of this study is to evaluate whether plasma levels of VEGF correlate with its immunohistochemical expression and with microvessel density (MVD) in patients with colorectal cancer. MATERIALS AND METHODS: Thirty-one patients with advanced colorectal cancer, who underwent surgery between February 1998 and April 2000, were included in this study. We measured the preoperative plasma levels of VEGF using the ELISA kit and immunostained the resected specimens for VEGF and CD34, as a marker of MVD. We then investigated the correlation among plasma levels of VEGF, expression of VEGF and MVD, and between these three factors and several clinical features. RESULTS: The plasma levels of VEGF were significantly associated with liver metastasis, the immunohistochemical expression of VEGF was significantly associated with lymph node metastasis and TNM stage, while MVD was significantly associated with lymph node metastasis, depth of invasion and TNM stage. Among the 3 parameters for angiogenesis studied, the plasma levels of VEGF significantly correlated with its immunohistochemical expression, and immunohistochemical expression of VEGF significantly correlated with MVD. There was no significant correlation between plasma levels of VEGF and MVD. CONCLUSION: Measuring plasma levels of VEGF is a good predictor of the immunohistochemical expression of VEGF in patients with advanced colorectal cancer, and may be a better indicator for tumor VEGF levels, since plasma levels can be measured much more quickly than expression levels. PMID- 12530026 TI - Relevance of c-erbB2, PLU-1 and survivin mRNA expression to diagnostic assessment of breast cancer. AB - The positivity rates of mRNA expression in breast cancer of the tumor-related genes for c-erbB2, PLU-1 and survivin are unclear. We quantitatively analyzed tissue samples from 39 breast cancers and non-cancerous parts of the same specimens for the above three mRNAs using a TaqMan reverse transcription polymerase chain reaction (RT-PCR). Using the mean + 2SD of non-cancerous sample as a cut-off value, the positivity rates of the tumors for c-erbB2, PLU-1 and survivin were 20.5%, 7.7% and 69.2%, respectively. Combining consideration of survivin with c-erbB2 and or PLU-1 increased the positivity ratio (survivin plus either of others, 76.9%; survivin plus both, 79.5%). Analysis by histological type indicated that survivin showed the highest positivity in ductal carcinoma and that survivin and PLU-1 showed the same positivity rate (40.0%) in the five carcinomas classified histologically as either solid-tubular or mucinous. Further, all cases that were positive for PLU-1 were negative for survivin. Survivin mRNA expression appeared more useful as a marker for diagnosis of breast cancer than c-erbB2 or PLU-1. However, PLU-1 appeared to vary independently of survivin, enhancing the usefulness of assays considering both in combination. PMID- 12530027 TI - Does P-glycoprotein-170 expression predict for chemoresistance in transitional cell carcinoma of the bladder? AB - INTRODUCTION: The glycoprotein P-170, causing drug efflux from the cells, may represent at least one cause of resistance to most drugs used in intravesical chemotherapy of superficial bladder cancer. MATERIALS AND METHODS: GP-170 was retrospectively assessed in 60 patients affected by superficial transitional cell tumours of the bladder. It was assessed by immunohistochemistry in a semiquantitative way by the intensity of staining and by the percentage of positive cells. Correlation of GP-170 expression with G-grade, T-category, multiplicity, recurrence rate and treatment was investigated. In 44 patients recurrence was analysed in relation to GP-170 basal expression and to its variations. The monoclonal antibody JSB1 (DBA) at 1:20 dilution was employed for the GP-170 assay. RESULTS: GP-170 expression increases with grade but was lower in multiple tumours. No difference between Ta and T1 categories was detected. GP 170 immunohistochemistry from different portions of the same tumour showed a very marked variability in 35.7% of patients. Seven patients (11.6%) were totally negative for GP-170. No statistically significant correlation was found between recurrence, progression and GP-170 basal expression. Similarly no correlation emerged between grade and stage variations at recurrence and modifications in GP 170 expression. One third of the tumours recurring after chemotherapy were negative for GP-170 in spite of an increase in recurrence rate and other risk factors. CONCLUSION: At the present stage of our experience, we have been unable to show that GP-170 is a useful marker for monitoring chemoresistance to intravesical chemotherapy in superficial bladder cancer. Furthermore, GP-170 determination has shown several technical difficulties. PMID- 12530028 TI - Molecular aspects of oral cancer. AB - Squamous cell carcinoma of the oral cavity is the sixth most frequent cancer in the world with approximately 30,000 new cases diagnosed annually in the United States. The overall 5-year survival rate for patients with oral squamous cell carcinoma is among the lowest for major cancers and has not changed during the past two decades. Despite therapeutic and diagnostic progress in oncology during the past decades, the prognosis of intraoral squamous cell carcinoma remains poor. Thus it seems that finding a biological tumor marker or tumor markers which will increase the early diagnosis and treatment monitoring rates, is of paramount importance in respect to improving prognosis. The latest studies pertaining to tumor markers and their role in the detection, diagnosis and treatment monitoring of oral squamous cell carcinoma are reviewed. The importance of cell surface markers including blood antigens, and growth factors and receptors, and intercellular markers including cytokeratins and AgNORs are discussed. Particular attention is paid to quantitative DNA (aneuploidy) and oncogenes. The pivotal tumor suppressor gene, p53, is dealt with in depth. PMID- 12530030 TI - Low p27Kip1 expression is associated with poor prognosis in oral squamous cell carcinoma. AB - BACKGROUND: p27Kip1 is a cyclin-dependent kinase inhibitor which regulates the progression of cells from the G1-into the S-phase in a cell cycle. Loss of p27Kip1 is associated with disease progression and an unfavorable outcome in several malignancies. The purpose of this study was to determine whether p27Kip1 expression can be a useful prognostic factor in oral squamous cell carcinoma (SCC) patients. PATIENTS AND METHODS: p27Kip1 expression was investigated by immunohistochemistry in tissue samples from 81 patients with oral SCC. The associations between p27Kip1 expression and clinicopathological characteristics and patient survival were also analyzed. RESULTS: Significant associations were found between p27Kip1 expression and histological grade (p = 0.010), therapeutic effect (p = 0.004) and patient outcome (p = 0.005). The 5-year survival rates of p27Kip1 high- and low-expression tumours were 80.4% and 56.7%, respectively, this difference being significant (p = 0.009) by log-rank test. Multivariate analysis revealed that reduced term survival was related to low levels of p27Kip1 expression (p = 0.008) and advanced stage (stages III and IV) (p = 0.003). CONCLUSION: These results suggest that reduction of p27Kip1 plays an important role in the progression of oral SCC and is considered to be a useful prognostic factor in oral SCC patients. PMID- 12530029 TI - Gemcitabine plus Epi-doxorubicin as first-line chemotherapy for bladder cancer in advanced or metastatic stage: a phase II. AB - Combination chemotherapy with newer, more active drugs in patients with advanced and/or metastatic bladder cancer might show improved response rate and survival. Gemcitabine (GEM) and Epidoxorubicin (EPI) have demonstrated activity in this disease. In addition, experimental studies in vitro have shown that the two agents have additive-synergistic effects when used in combination. Our prior phase I dose-finding study in previously untreated patients with advanced or metastatic bladder cancer defined recommended doses for further trials of GEM 1000 mg/m2 and EPI 25 mg/m2 on days 1, 8 and 15 every 28 days. A phase II trial at this dose level was initiated in previously untreated patients to assess efficacy and toxicity. Eligible patients had measurable disease; Karnofsky performance status (PS) of > 40; no prior chemotherapy; and adequate bone marrow reserve, cardiac, hepatic and renal function. Thirty- one patients (22 males, 9 females) with median age of 64 (range 44-75) and median PS of 80 were accrued, and all were eligible. Twelve patients had T4N1-2 M0, 8 had lymph node only metastases, while 11 had visceral metastases (liver, bone, lung). A total of 181 cycles was administered (range 3-7 per patient). Major toxicities (WHO grade > or = 3) were: neutropenia in 5 patients, thrombocytopenia in 2 patients, and anemia in 2 patients. Three patients had febrile neutropenic episodes and only 3 patients required dose reduction. Grade 1-2 non-hematological toxicities included nausea/vomiting, stomatitis and alopecia. No cardiac toxicity was observed. Of the 30 response evaluable patients, 17 (57%) demonstrated a major response (3 complete and 14 partial) (95% CI: 39%-75%), 7 had stable disease (23%) and 6 progressed (20%). These preliminary results confirm the phase I observation that the combination of GEM--EPI is highly active in the treatment of advanced and metastatic bladder cancer with a favourable toxicity profile. PMID- 12530031 TI - Expression of tissue factor is associated with clinical features and angiogenesis in prostate cancer. AB - BACKGROUND: Tissue factor (TF), the main initiator of blood coagulation, is involved in cancer metastasis and progression. We examined the role of TF on prostate cancer. MATERIALS AND METHODS: Immunohistochemical analysis was performed using the anti-TF antibody. Intra-tumoral blood vessels were visualized by staining endothelial cells with CD34 antibody. We examined the expression of TF and the microvessel density (MVD) in 66 biopsy specimens of prostate cancer, in order to investigate the relationship between the expression of TF and the clinicopathology of prostate cancer. RESULTS: TF antigen was positive in 41 (62%) of the specimens. There were significant differences in TF expression according to the pretreatment prostate specific antigen (PSA) level (p = 0.0193) and bone metastasis (p = 0.0029). MVD was significantly related to bone metastasis (p = 0.0175). TF-positive carcinomas more frequently presented high MVD expressions (p = 0.017) than TF-negative tumors. CONCLUSION: These results suggest that increased angiogenesis associated with TF expression might cause the metastasis and progression of prostate cancer. PMID- 12530032 TI - Prognostic value of uPA, PAI-1 and PAI-2 mRNA expression in primary breast cancer. AB - BACKGROUND: The prognostic value of uPA, PAI-1 and PAI-2 mRNA expression was studied in a retrospective series of 130 primary breast cancer patients (median follow-up 8.1 years). MATERIALS AND METHODS: UPA, PAI-1 and PAI-2 mRNA were quantified by means of real-time quantitative RT-PCR. Comparison with the corresponding protein levels determined by ELISA was performed in 21 cases. RESULTS: Higher uPA protein values were found in cases with high mRNA values, but the relationship was of borderline significance. PAI-1 and PAI-2 mRNA were positively correlated to protein values. The mRNA expression of uPA, PAI-1 and PAI-2 was significantly correlated with one another. Higher uPA and PAI-1 mRNA values were significantly associated with shorter disease-free survival (DFS) (p = 0.002; p = 0.03, respectively). Low and very high PAI-2 mRNA values tended to be associated with longer DFS. In Cox multivariate analyses, higher uPA and PAI-1 mRNA values were independently associated with shorter DFS. PAI-2 was not retained as a significant variable in the Cox model. CONCLUSION: These preliminary results indicate that, in breast cancer, uPA, PAI-1 and PAI-2 mRNA analysis by quantitative RT-PCR give results comparable to those obtained at the protein level. PMID- 12530033 TI - Breast cancer in Nigeria and Finland: epidemiological, clinical and histological comparison. AB - BACKGROUND: We compared the histology and patterns of occurrence of breast cancers in Nigeria (n = 297) and Finland (n = 285). PATIENTS AND METHODS: The histology of invasive ductal carcinoma (IDC) was re-evaluated using similar criteria. The clinical data were extracted from medical records. RESULTS: The mean age at presentation was 42.7 (12.2) years in Nigeria vs. 58.7 (12.5) years in Finland. In both populations there was an association between reproductive factors and the occurrence of breast cancer. In Nigeria, 53.2% of cases belonged to stages 3 and 4 (vs. 6.7% in Finland). In Finland there were higher frequencies of lobular, tubular and mucinous types than in Nigeria. The Nigerian material had more medullary type (2.7% vs. 0.7% in Finland), extensive necrosis, nuclear atypia and pleomorphism, with coexisting pleomorphic ductal carcinoma in situ. At 2 years after treatment, the survival figures for Nigeria and Finland were 72.8% and 96.4%, respectively. CONCLUSION: The histology of Nigerian and Finnish cancers clearly differ. Nigerian cancers appear more advanced with higher grade. The atypical in situ component is clearly more common in Nigeria. Part of the differences can be explained by diagnostic and treatment delays associated with misguided socio-cultural beliefs, poor health care access and impaired immunity. PMID- 12530034 TI - Intermediate dose pre-operative irradiation for carcinoma of the rectum. AB - BACKGROUND: To assess the locoregional control, survival and toxicities of combined intermediate-dose pre-operative irradiation and surgery for rectal cancer patients. MATERIALS AND METHODS: Forty-two patients with adenocarcinoma of the rectum underwent intermediate-dose pre-operative radiotherapy. The median age was 58 years. the prescribed dose was 30.6 Gy to the whole pelvis, while a boost of 4 x 3 Gy to the primary tumor was added. The calculated linear-quadratic dose was 52 Gy. RESULTS: Failure as a first site was pelvic in 2 and distant in 11. The crude pelvic failure rate was 9.5%. The 3- and 5-year overall survival was 75% and 63%, respectively. Pre-operative radiotherapy was well-tolerated by all. No anastomotic breakdown was seen. Nor was any death attributable to any adverse effects of the treatment. CONCLUSION: The present results suggested that pre operative intermediate-dose radiotherapy is effective in preventing pelvic recurrence in rectal cancer patients with acceptable toxicities. PMID- 12530035 TI - Differential diagnosis of keratoacanthoma and squamous cell carcinoma of the epidermis by MIB-1 immunohistometry. AB - OBJECTIVE: For the analysis of cellular proliferative activity, the MIB-1 immunopositivity of keratoacanthoma (KA, n = 49), squamous cell carcinoma (SCC, n 48) and each of four cases diagnosed probably as KA or probably as SCC were analyzed by means of immunohistometry. STUDY DESIGN: Immunohistochemical reactions were performed on 3-micron sections from routinely formalin-fixed and paraffin-embedded surgical specimens, using an indirect peroxidase method. The rate of immunostained cells was determined using a TV-image analysis system CM-2 (Hund, Germany). Twenty viewing fields (0.97 mm2) were measured with 20:1 objective magnification. An average of 1688 cells were assessed in each case. RESULTS: The mean MIB-1 immunopositivity (MIB-1mean) was higher in SCC (42.3% +/- 19.1%) than in KA (26.8% +/- 9.8%). The distribution of the single values differed significantly (p = 0.0002). To test the suitability of MIB-1 immunohistometry for the differential diagnosis between KA and SCC, various thresholds were investigated. Using a threshold of 30%, SCC can be detected with a sensitivity of 70.8% (34 out of 48) and a specificity of 67.3% (33 out of 49). If a specificity of > or = 85% is required (42 out of 49, 85.7%), the sensitivity of the test decreases to 56.3% (27 out of 48) based on a threshold of 37.5%. Using the MIB-1 value of the most positive focus of the lesion (MIB-1max), the results were of minor significance; at a specificity level of > or = 85% (42 out of 49, 85.7%) a sensitivity rate of only 43.8% (21 out of 48) could be obtained (threshold: 75%). CONCLUSION: As some overlap of the single values has to be considered, MIB-1 immunohistometry, although presenting new insights into the proliferative potential of KA and SCC, is of only limited value for the differential diagnosis of the two lesions in routine surgical pathology. PMID- 12530036 TI - High molecular weight DNA fragmentation assay: a sensitive method for monitoring in vivo response to antileukemic chemotherapy. AB - A large number of antineoplastic agents exert their cytotoxic effects through induction of apoptosis. However, despite the availability of an increasing number of methods to identify apoptosis, attempts to detect apoptosis in clinical samples are rarely successful. Here we report the reliability of the high molecular weight (HMW) DNA fragmentation assay in monitoring the in vivo response of a patient diagnosed with B-CLL to fludarabine (F-ara-A) infusion monotherapy by tracing 50 Kb DNA fragments in his peripheral blood at different time-points before and after infusion. The HMW DNA fragmentation assay proved qualitative and quantitative sensitivity in following the progression of apoptosis in the patient's circulating population of leukemic blasts while, under the same circumstances, conventional cytometric parameters of apoptosis remained negative. PMID- 12530037 TI - Histopathology of the cardiac conduction system in a case of metastatic pancreatic ductal adenocarcinoma. AB - Metastatic tumor involving the heart occasionally complicates the course of patients with neoplasia. A 70-year-old white male died suddenly. Post-mortem histological examination showed the presence of cardiac metastatic pancreatic ductal adenocarcinoma compressing the cardiac conduction system, which had led to motility, hemodynamic and conductive abnormalities deteriorating into sudden death. PMID- 12530038 TI - Clinical usefulness of chemosensitivity test for advanced colorectal cancer. AB - We assessed the usefulness of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay in the evaluation of appropriate adjuvant cancer chemotherapy for patients with advanced colorectal cancer. We analyzed 405 cases of colorectal cancer treated between January 1990 and August 1999 in terms of the MTT assay and survival outcome. Patients with Dukes' C and D were classified into a "surgery alone" group (n = 53), a "sensitive" group who received drugs that had a greater than 50% inhibition rate by MTT assay (n = 23), or "resistant"" group who were insensitive to the chemotherapy drugs (n = 124). Statistically significant differences in survival outcome were observed between the groups, with the sensitive group showing significantly better survival compared with the resistant group (p = 0.0158) and the surgery-alone group (p = 0.0004). Our results suggest that the MTT assay may be useful in evaluating the optimum adjuvant chemotherapy for patients with advanced colorectal cancer. PMID- 12530039 TI - Gemcitabine plus cisplatin in the treatment of patients with advanced non-small cell lung cancer: a phase II study. AB - BACKGROUND: Lung cancer is the leading cause of cancer death in men worldwide; most cases are not suitable for radical surgery at diagnosis and palliative treatment remains the primary goal of therapy. Cisplatin and gemcitabine are among the most active cytotoxic agents for the treatment of non-small cell lung cancer (NSCLC): they have non-overlapping toxicity and preclinical studies have demonstrated their potential synergistic interaction. PATIENTS AND METHODS: The aims of the present study were to assess the activity and tolerability of cisplatin 80 mg/m2 on day 1, combined with gemcitabine 1000 mg/m2 on days 1 and 8, administered every 3 weeks. A total of 46 consecutive patients with advanced NSCLC entered this study; all of them were evaluable for toxicity and for activity. RESULTS: According to an intent-to-treat analysis, 15 patients attained a partial response (33%), 9 (20%) obtained a disease stabilisation and 22 (47%) progressed. This regimen appeared to be modestly toxic, with grades 3-4 leukopenia and thrombocytopenia observed in 10% and 6% of cases respectively; grade 3 vomiting appeared in 12 patients (26%) and grade 3 mucositis in 1 patient. The median time-to-progression and overall survival were 200 and 400 days, respectively. CONCLUSION: Our study of gemicitabine + cisplatin on stage IV NSCLC patients achieved favourable results in terms of toxicity and overall survival. PMID- 12530040 TI - Interleukin-2, interferon-alpha and medroxyprogesterone acetate in metastatic renal cell carcinoma. AB - BACKGROUND: Interleukin-2 (IL-2) and interferon-alpha (IFN-alpha) are the main immuno-biological agents used in the therapy of metastatic renal cell carcinoma (RCC). Unfortunately the promising results obtained in biological studies have not yet been confirmed in clinical studies. One reason is linked to the immunosuppression of metastatic patients which is caused by macrophage products. IL-6 in particular is considered a growth factor for RCC. Medroxyprogesterone acetate (MPA) may interfere with IL-6 macrophage production, possibly causing a synergistic effect in association with IL-2 and IFN-alpha. Therefore the purpose of our study was to evaluate the toxicity and the efficacy of the association between IL-2, IFN-alpha and MPA. PATIENTS AND METHODS: Forty-two consecutive patients with metastatic RCC were enrolled. IL-2 was administered subcutaneously at doses of 4.5 million UI on days 1-5, 8-12, 15-19 and 22-26; IFN-alpha was administered s.c. at a dose of 3 million t.t.w; MPA was administered orally at a dose of 1000 mg daily. This schedule was repeated after a rest of 2 weeks. RESULTS: Toxicity was mild: the main symptoms observed were fatigue and fever. Six CR (14%), five PR (12%), thirteen SD (31%) and seventeen PD (41%) were observed for an overall response rate of 26%. Patients with good PS and low levels of CRP had a better prognosis. CONCLUSION: Considering both the good activity and the low toxicity of this scheme, we think that it could be carried out in normal clinical practice. PMID- 12530041 TI - Gemcitabine as single agent chemotherapy in elderly patients with stages III-IV non-small cell lung cancer (NSCLC): a phase II study. AB - INTRODUCTION: In the past the unfavourable profile of toxicity of antineoplastic drugs employed (i.e. cisplatinum) weakened the role of chemotherapy in aged patients with non-small cell lung cancer (NSCLC). Recently, new active drugs with lesser toxicity have became widely used in this setting. In this prospective study we evaluated the efficacy and tolerability of gemcitabine in elderly patients with stages III-IV NSCLC. MATERIALS AND METHODS: From December 1996 to April 1999, we enrolled 52 previously untreated elderly patients with advanced/metastatic NSCLC. Gemcitabine was administered at 1000 mg/mq i.v. over 30 minutes weekly for three consecutive weeks every 28 days. The planned number of cycles was three while responding or stable patients received chemotherapy until disease progression or the ninth cycle. A total of 291 cycles were delivered with a median number of 6 cycles (range: 3-9). An evaluation of the quality of life was performed every three courses of gemcitabine. RESULTS: After three cycles of treatment, a complete response was seen in four patients (7.7%), partial response in 16 patients (30.8%) with an overall response rate of 38.5%. Nineteen patients (36.5%) showed stable disease, while thirteen patients (25%) progressed. Median progression-free survival was 22 weeks, median duration of response 26 weeks, median overall survival 34 weeks and 1-year overall survival 46.1%. A statistically significant improvement in the quality of life was registered only after the first three cycles of gemcitabine (p = 0.045). Toxicity was extremely mild. CONCLUSION: In elderly patients with stages III-IV NSCLC gemcitabine showed good activity with a mild toxicity and could be considered a valid therapeutic option in this setting. PMID- 12530042 TI - Intrathecal chemotherapy in carcinomatous meningitis from breast cancer. AB - Meningeal metastases occur in 2-3% of patients with breast cancer, leading to neurological morbidity and increased mortality. The criteria for treatment choice are controversial and intrathecal chemotherapy (ITC) has no documented role in the management of this disorder. We therefore evaluated the efficacy of an ITC regimen for patients presenting with carcinomatous meningitis from breast cancer. PATIENTS AND METHODS: Patients with meningeal carcinomatosis with or without concomitant parenchymal brain metastasis, were treated with repeated courses of intrathecal chemotherapy according to the following alternated weekly schedule: Day 1: Thiotepa 10 mg, methotrexate 15 mg, hydrocortisone 30 mg; Day 5: cytarabine (Ara-C) 70 mg, methotrexate 15 mg, hydrocortisone 30 mg. Folinic acid 15 mg was given orally, every six hours after methotrexate on days 2-3 and 6-7. RESULTS: Thirteen consecutive patients were treated. The median age was 45 (range 30-67) years. Eleven patients had performance status (PS) 2-3. Nine patients had other metastatic sites; synchronous parenchymal brain metastasis were present in 5 patients. Concomitant systemic chemotherapy was administered in 5 patients and external whole brain radiotherapy in 7 patients. With 12 evaluable patients we observed no responses or improvement in symptoms. Side-effects were minimal. CONCLUSION: In our series of patients, ITC failed to provide objective response or relief in clinical symptoms. Despite evidence reported in the literature indicating symptomatic improvement after ITC in a number of patients with leptomeningeal metastasis, the results of our study confirm the controversial role of ITC. New drugs and new modalities of treatment should be studied in order to efficiently control meningeal involvement of breast cancer. PMID- 12530043 TI - Clinical significance of soluble c-erbB-2 levels in the serum and urine of patients with gastric cancer. AB - BACKGROUND: Both tissue c-erbB-2 expression and serum levels the shed protein have been shown to correlate with tumour stage in a range of adenocarcinomas. This study measured serum and urinary c-erbB-2 levels in patients with gastric cancer, assessing their role in cancer-specific survival and the effects of resectional surgery. PATIENTS AND METHODS: Serum and urinary c-erbB-2 concentrations were measured with commercial enzyme-linked immunosorbent assay in 41 healthy controls and in 54 gastric cancer patients. Serum and urinary c-erbB-2 levels in cancer patients were determined before and 7 days following tumour surgery. RESULTS: Preoperative serum and urinary c-erbB-2 levels in gastric cancer patients were significantly higher than those in controls although there were no significant associations between these levels and tumour pathology. Serum c-erbB-2 levels decreased significantly after radical resection of the primary tumour and were an independent prognostic factor for survival, whereas there were no changes in urinary c-erbB-2 levels after surgery or an association with patient survival. CONCLUSION: Gastric cancer patients show higher serum and urinary c-erbB-2 levels compared to healthy controls. Preoperative serum c-erbB-2 concentration decreases significantly after radical resection of the primary tumour and is an independent prognostic factor for patient survival. PMID- 12530044 TI - Experience with the Advanced Breast Biopsy Instrumentation (ABBI) system. AB - BACKGROUND: The Advanced Breast Biopsy Instrumentation (ABBI) System is designed to excise nonpalpable breast lesions under stereotactic control. We report our experience with special regard to the histological evaluation of margins. PATIENTS AND METHODS: Breast biopsies using the ABBI system were performed on 101 patients with microcalcifications. In histologically-proven breast cancer, a re excision was performed. RESULTS: Malignant lesions were found in thirteen patients (3 CLIS, 5 DCIS, 5 invasive ductal carcinoma). The margins were positive in two specimens with DCIS. In subsequent lumpectomies one patient with invasive cancer had residual intraductal cancer. All the patients with DCIS had residual cancer, even those with negative margins of the ABBI-specimen. Only minor complications were observed with the ABBI procedure. CONCLUSION: The ABBI system is a safe, minimally invasive stereotactic breast biopsy technique. It saves open biopsies in atypical hyperplasia and CLIS. In cases of DCIS or invasive cancer re excision is inevitable. PMID- 12530045 TI - Tolerability of Raltitrexed ('Tomudex') in elderly patients with colorectal cancer. AB - PURPOSE: Colorectal cancer (CRC) is one of the major health problems of the Western world and the proportion of elderly patients with CRC is growing. Raltitrexed ('Tomudex'), a specific thymidylate synthase inhibitor, has shown efficacy and manageable toxicity in elderly CRC patients. In this retrospective study, the tolerability of raltitrexed in patients with CRC was examined in relation to age. PATIENTS AND METHODS: Toxicity parameters, graded according to World Health Organization criteria, were assessed in two patient groups: < 70 and > or = 70 years old. In total, 56% (50 out of 90) of patients treated with raltitrexed (3 mg/m2 as a 15-minute intravenous infusion every 3 weeks) were aged > 70 years (M:F 28:22; Eastern Cooperative Oncology Group performance status 0 1:2 38:12). RESULTS: Overall, 437 cycles of chemotherapy were administered and grade 3-4 toxicity was reported in < 10% of patients. There were no clinically significant differences between the two age groups, apart from grade 3-4 asthenia, which was reported by 6% and 0% of patients aged > or = 70 and < 70 years, respectively. This was in spite of a significantly lower calculated mean creatinine clearance in patients aged > or = 70 years compared with those patients < 70 years of age. CONCLUSION: The raltitrexed toxicity profile does not appear to be significantly influenced by age; however caution is recommended in the management of elderly patients, particularly in the presence of impaired renal function. PMID- 12530046 TI - Unique pineal gland metastasis of clear cell renal carcinoma: case report and review of the literature. AB - The metastatic involvement of the pineal gland is an extremely unusual event; it has a 4% incidence in patients with disseminated neoplasias. Most metastatic pineal lesions are asymptomatic. Only in a small number of cases the symptoms produced by metastatic involvement of this organ precede those of the primary tumor or those of another metastatic site. To our knowledge the herein reported case is the first in which the pineal gland was apparently the unique metastatic site of a primitive kidney carcinoma and where the symptoms produced by metastasis in the pineal region were the first sign of the disease. PMID- 12530047 TI - Correlation of 99mTc-MIBI brain spect (functional index ratios) and survival after treatment failure in malignant glioma patients. AB - AIM: This preliminary study was conducted to determine the association between 99mTc-SESTAMIBI (MIBI) brain SPECT and survival in malignant glioma patients after treatment failure. MATERIALS AND METHODS: Twenty-five malignant glioma patients with clinical deterioration were studied. Tomoscintigraphy was performed 15 minutes after intravenous injection of 1110 Mbq MIBI. The images were obtained from a dual head gamma camera using fan beam collimators. Transverse, coronal and sagittal views were reconstructed. Maximum uptake of MIBI in the lesion was expressed as a ratio to that in the controlateral hemisphere. RESULTS: A MIBI uptake was found in all 25 patients. This uptake was correlated to tumour recurrence proved by histological fragments and/or the rapid fatal evolution of these patients. From scintigraphic parameters, two groups of patients were noted. Group 1 (20 patients) had a significantly better survival time (8.25 months) than did Group 2 (5 patients) (2.9 months). Tumour volume predicted survival (group 1 vs group 2, p < 0.0006). A significant correlation between the tumour/background ratio and survival time for each group was observed. CONCLUSION: A correlation between functional index ratios and the time of survival of patients was noted. The functional index ratios of brain SPECT are in accordance with the potential aggressiveness of the recurrent malignant glioma. PMID- 12530048 TI - A new cisplatin/gemcitabine schedule in locally advanced (IIIB) and metastatic (IV) non-small cell lung cancer: relationship between dose-intensity and efficacy. A phase II study. AB - BACKGROUND: Cisplatin/gemcitabine are one of the "standard" chemotherapy schedules in locally advanced and metastatic NSCLC cancer. A number of trials documented that omission of gemcitabine on day 15 and reduction of cisplatin up to 70 mg/mq are equivalent in term of response rates to "classic" administrations on days 1, 8 and 15 with cisplatin 100 mg/mq. The aim of this study was to confirm this evidence and to demonstrate that a further reduction of gemcitabine dose-intensity may be performed with the same efficacy on response. PATIENTS AND METHODS: Fifty untreated patients with locally advanced and metastatic NSCLC entered the study: 24 stage IIIB and 26 stage IV. The median age was 65 years (range 32-76); 44 males and 6 females Genicitabine was administered 1000 mg/mq weekly on days 1 and 8 followed by a 2-week rest and cisplatin 80 mg/mq on day 2 of each 28-day-cycle. RESULTS: Forty-five patients were evaluable for response and all for toxicity. The overall response rates were 35.5% with 16 partial responses (95% Confidence Interval: 32%-61%). Most of the objective responses were seen in IIIB patients (56% of the stage IIIB and 44% of the stage IV patients responded). According to the intent-to-treat-principle, the response rates were 32% (16 out of 50 patients). The median dose-intensity of gemcitabine and cisplatin was respectively 477.6 mg/mq/week (481.4 for responders) and 19.5 mg/mq/week (19.9 mg/mq for responders). The median response duration was 5 months (range 1-18) and the median time to progression was 5 months (1-21); median survival was 9 months (range 2-31). The main toxicity was haematological: thrombocytopenia grade IV in 5 patients (10%) and grade III in 11 patients (22%); neutropenia grade III-IV in 4 patients (8%); grade III anemia in 3 (6%). Asthenia was the most significant non-haematological toxicity and was observed in 19 patients (38%). CONCLUSION: This trial confirmed the efficacy of a schedule with 2 administrations of gemcitabine (on days 1, 8) and a cisplatin dose on day 2 lower than 100 mg/mq. Moreover, the same efficacy was obtained with a median-dose intensity of cisplatin and gemcitabine lower than planned in a 21-day-schedule. For safety and low toxicity, we think that this schedule provides another chance to treat patients with non-small cell lung cancer, especially the elderly or patients with coexistent medical illnesses. PMID- 12530049 TI - The significance of Tenascin-C serum level as tumor marker in squamous cell carcinoma of the head and neck. AB - BACKGROUND: Tenascin, an extracellular matrix glycoprotein, is transiently present in embryonic tissue, in benign granulation tissue, but also in several highly anaplastic tumors like fibrosarcoma, melanoma and squamous cell carcinoma of the skin. This study was performed to validate elevated Tenascin serum levels as a possible marker for head and neck squamous cell carcinomas (HNSCC). PATIENTS AND METHODS: Tenascin serum levels were evaluated in patients with primary (n = 92) and with recurrent (n = 28) HNSCC. Patients with benign, non inflammatory ear, nose and throat diseases (n = 16) served as the control. The Tenascin serum levels were measured by ELISA (Aventis). RESULTS: Serum Tenascin concentrations of patients with benign ENT diseases ranged between 0.37 and 2.19 micrograms/ml (n = 16, mean +/- SD: 1.23 +/- 0.59 micrograms/ml), of patients with HNSCC (primary diagnosis) between 0.05 and 8.75 micrograms/ml (n = 92, mean +/- SD: 1.81 (1.36 micrograms/ml) and of patients with recurrent HNSCC between 0.53 and 10.0 micrograms/ml (n = 28, mean +/- SD: 2.78 +/- 2.2 micrograms/ml). CONCLUSION: We found a significant elevation of Tenascin serum levels only in patients with higher tumor stages (T4/UICC4) (p < 0.01/p < 0.1) or recurrent disease compared to Tenascin serum levels in healthy controls. Thereby Tenascin serum levels cannot be used clinically as a routine serum marker for the control of head and neck cancer. Further investigations are necessary to evaluate whether the measurement of Tenascin levels as tumor markers could offer additional information to the clinical outcome of patients with HNSCC. PMID- 12530050 TI - Application of cisplatin as intraoperative hyperthermic peritoneal lavage (IHPL) in patients with locally advanced gastric cancer: analysis of pharmacokinetics and of nephrotoxicity. AB - The purpose of this study was to assess the extent of the systemic absorption of cisplatin during intraoperative hyperthermic peritoneal lavage (IHPL) in patients with locally advanced gastric cancer. MATERIALS AND METHODS: The pharmacokinetics and nephrotoxicity of cisplatin were analyzed in patients receiving IHPL (8000 ml of Ringer's solution containing 150 mg/m2 cisplatin and 15 mg/m2 mitomycin C for one hour at 43.5 degrees C). Levels of ultrafiltrable platin were determined by flameless atomic absorption spectrometry. Nephrotoxicity was assessed by nephelometric analyses of urinary marker-proteins. The data were compared to respective analyses in patients receiving intravenous cisplatin. RESULTS: Twenty four patients received five applications of cisplatin as IHPL (five patients) and 53 applications of intravenous cisplatin (21 patients). Platin levels within the lavage fluid declined monophasically (half-life, 0.48 +/- 10 hours; area under curve (AUC) 29,274 +/- 9075 ng/ml*h). The pharmacokinetic parameters calculated for IHPL vs. intravenous application of cisplatin were: maximum plasma levels 2392 +/- 407 vs. 1349 +/- 692 ng/ml; terminal half-lives 93 +/- 73 vs. 36 +/- 9 hours; AUC 9508 +/- 856 vs. 11,627 +/- 3372 ng/ml*h; total urinary excretion of platinum 24 +/- 6 vs. 49 +/- 13% of dose; renal clearance 127 +/- 34 vs. 145 +/- 35 ml/min. Pathologic urinary albumin excretion occurred on days 9 +/- 0 vs. 5 +/ 2 (maximum 232 +/- 179 vs. 20 +/- 20 mg/l). Plasma creatinine levels rose to 1.5 +/- 0.4 vs. 0.9 +/- 0.1 mg/dl on days 15 +/- 4 vs. 16 +/- 26. The degree of albuminuria was related to the clearance of platin from the lavage fluid (p = 0.048). CONCLUSION: A significant amount of intraperitoneally applied cisplatin is available systemically and probably adds to the nephrotoxicity of IHPL. PMID- 12530051 TI - Multifocality of renal cell tumors is a factor to consider before performing a partial nephrectomy. AB - BACKGROUND: We conducted a retrospective analysis of radical nephrectomies performed in our clinic in order to evaluate the incidence of multifocality in relation to the stage, grade, size and histological type. MATERIALS AND METHODS: Preoperative imaging studies and the final pathological results of surgical specimens were reviewed. Separate primary tumors at a distance of more than one cm from the primary tumor were counted. Of 56 renal cell cancers 10 (17.8%) were multifocal. Of these 10, 3 were found with primary tumors less than or equal to 4 cm, while 7 were found with tumors between 4-10 cm. Three multiflocal tumors were found with TI tumors, none with T2, 4 with T3a and three with T3b primary tumors. Six cases were present where the histological subtype of the largest tumor was clear cell, while 4 had mixed cell histology. No multifocality was observed in the 4 grade I tumors, while 3 of the 32 grade II tumors and 6 of the 20 grade III tumors were multifocal. The only grade IV tumor presented with a separate primary tumor. Statistically significant correlation was found between multifocality and tumor grade stage and clear cell histology subtype. There was no correlation between the primary tumor size and the possibility of multifocality. CONCLUSION: Although partial nephrectomies are justified from their survival rates, close follow-up is necessary because separate primary tumors may still exist in the kidney, left behind, irrespective of tumor size. PMID- 12530052 TI - Sentinel nodes outside level I-II of the axilla and staging in breast cancer. AB - BACKGROUND: The aim of the study was to evaluate the incidence of sentinel nodes and sentinel node metastases outside levels I-II of the axilla in breast cancer. PATIENTS AND METHODS: Altogether 170 breast cancer patients with 172 clinically node-negative T1-T2 tumours underwent lymphoscintigraphy and were included in a prospective study. RESULTS: The lymphoscintigraphy showed sentinel node(s) in the axilla in 150 (87%) breast cancer cases. Thirty (17%) patients had sentinel nodes outside the axilla. Lymphatic drainage solely outside the axilla was encountered in two patients. Lymph node metastases were found in the axilla in 40% and outside the axilla in 17% of the 30 patients with extra-axillary sentinel nodes. Two patients with sentinel node metastases outside the axilla had no axillary metastases. CONCLUSION: The biopsy of sentinel nodes outside the axilla is a potential tool for more accurate staging in breast cancer, since it provides additional information as compared to axillary staging alone. PMID- 12530053 TI - Thymidylate synthase expression in rectal cancer and proliferation, assessed by cyclin A and Ki-67 expression. AB - BACKGROUND: Levels of the enzyme thymidylate synthase (TS) are of prognostic significance in colorectal cancer. It may be argued that the levels of TS merely reflect the proliferative activity and could be replaced by markers of proliferation. MATERIALS AND METHODS: We used immunohistochemical approaches to examine the expression of TS, Cyclin A and Ki-67 in morphologically well-defined tumor areas in consecutive slices of rectal cancer, using the antibodies TS 106, NCL-Cyclin A and Mib-1. RESULTS: There was a linear relationship between Cyclin A and Ki-67-positive cells (p < 0.0001). There did not seem to be any significant relationship between TS-expression and the frequency of Cyclin A-positive cells (p = 0.1) but a significant correlation was observed between TS-expression and the frequency of Ki-67-positive cells (p = 0.02). CONCLUSION: TS, immunohistochemically-detected in rectal cancer cells, is not associated with an accurate assessment of the proliferative stage. The prognostic value of TS determination can only partly be explained by the proliferative activity. PMID- 12530054 TI - Galectin-3 expression alters adhesion, motility and invasion in a lung cell line (DLKP), in vitro. AB - BACKGROUND: Galectin-3, a beta-galactosidase-binding protein, is involved in regulating many physiological and pathological cellular processes. The significance of galectin-3 in human lung and nasal carcinoma cells has not yet been elucidated. MATERIALS AND METHODS: Using RT-PCR and Western blotting techniques, the constitutive level of galectin-3 in the human non-small cell lung carcinoma cell line, DLKP, was investigated. Following galectin-3 cDNA transfection into these cells, growth, toxicity, adhesion, motility and invasion assays were used to investigate the relevance of galectin-3 over-expression. RESULTS: Galectin-3 over-expression did not induce a multi-drug resistance phenotype or significantly affect cell growth rate, but it did result in enhanced (i) adhesion to extracellular matrix components; (ii) cell motility; and (iii) in vitro invasiveness. Furthermore, studies of RPMI-2650 variants suggest that galectin-3 expression correlates with nasal carcinoma cell invasiveness. CONCLUSION: Our results suggest that galectin-3 expression levels in both lung and nasal tumour cells may play a role in cell motility, invasion, and metastasis. PMID- 12530056 TI - Head and neck squamous cell carcinoma is highly sensitive to vinorelbine in Vitro. AB - The vinorelbine sensitivity of eight recently established head and neck squamous cell carcinoma (SCC) cell lines was tested using the 96-well plate clonogenic assay. The chemosensitivity of these head and neck SCC cell lines to vinorelbine expressed as IC50, corresponding to the drug concentration causing 50% inhibition in clonogenic survival, varied between 0.6 and 1.0 nM. The dose-dependent growth inhibition caused by vinorelbine was measured in three of these cell lines. A clear growth inhibition was observed at a concentration of 3 nM. The same cell lines were studied with flow cytometry. When exposed to 3 nM and 5 nM vinorelbine, an accumulation of the cells in the G2/M-phase was observed in all cultures after 12 hours. The morphological changes induced by 3 nM and 5 nM vinorelbine to the UT-SCC-33 cell line were analysed with time-lapse video microscopy. In the cultures treated with 5 nM vinorelbine, the cells stayed mitotically arrested for 2-32 hours and thereafter died morphologically by apoptosis. These results indicate that, in vitro, the head and neck SCC is consistently sensitive to vinorelbine, which blocks the cell cycle in G2/M, the most radiosensitive phase. These encouraging results suggest that vinorelbine may potentially be used in conjunction with radiotherapy in the treatment of head and neck SCC. PMID- 12530055 TI - Effects of the isoprenoids perillyl alcohol and farnesol on apoptosis biomarkers in pancreatic cancer chemoprevention. AB - Perillyl alcohol, farnesol and geraniol have chemotherapeutic activity toward pancreatic and other cancers. Perillyl alcohol induces apoptosis and increases expression of the proapoptotic protein Bak in cultured pancreatic tumor cells. We tested the hypothesis that farnesol and geraniol would have similar effects. After 48 hours of treatment with farnesol geraniol or perillyl alcohol, human BxPC3 pancreatic cancer cells exhibited a 3 to 10-fold increase in apoptosis and higher Bak expression than the controls. We then tested the hypotheses that perillyl alcohol and farnesol would have chemopreventive activity toward pancreatic cancer and would increase Bak expression and apoptosis in vivo. Hamster pancreatic cancer was initiated at time 0 with N-nitrosobis(2 oxopropyl)amine. Animals were fed control, 2% (w/w) perillyl alcohol, or 1% (w/w) farnesol diets from weeks 5-42. Pancreatic carcinoma incidence was decreased by perillyl alcohol and farnesol. Hyperplastic pancreatic ductal neoplasms from perillyl alcohol and farnesol-treated animals had higher Bak protein expression (p < 0.05), and somewhat higher apoptotic rates, diminished expression of the antiapoptotic protein BCL-XL, and lower rates of DNA synthesis than the controls. PMID- 12530057 TI - Combined effect of hyperthermia at 42 degrees C and irradiation dose of 2 Gy on two rat yolk sac tumor cell lines with different radio-thermosensitivity in vitro. AB - BACKGROUND: Two rat yolk sac tumor cell lines, NMT-1 and NMT-1R, are of the same origin and of different sensitivity to irradiation and to heat. The aim of this study was to investigate the sensitivities of these two cell lines to combined treatments of low-dose irradiation at 2 Gy and hyperthermia at 42 degrees C. MATERIALS AND METHODS: The cell survival was assayed by soft agar clonogenic assay. After the survival curves of radiation alone and of heat alone at various temperatures were estimated, not only the effect of irradiation on heat, but the effect of heat on irradiation were evaluated with sequential treatments in both cell lines. These effects on survival curves were evaluated by the enhancement ratios at isosurvival levels of 37%, 10% and 1%, respectively. RESULTS: NMT-1 was more sensitive to radiation but more resistant to heat than NMT-1R. For 1% survival level, radiosensitivity in NMT-1 was 1.32 times that in NMT-1R, while thermal sensitivity at 42 degrees C in NMT-1R was 2.73 times that in NMT-1. For sequential treatment, thermosensitization by a radiation dose of 2 Gy in radiosensitive NMT-1 was greater than that in radioresistant NMT-1R. Following heat at 42 degrees C for 1 hour, increased radiosensitivity in NMT-1R was significant, whereas the same heat treatment produced an increase in the radiation sensitivity of NMT-1 with a reduction of the survival curve shoulder but with less slope modification. There was no difference in the surviving fraction in the time-course of a combination of heat and irradiation at various intervals within 6 hours for NMT-1 except for heating immediately after irradiation. However a significant increase in survival was observed when heat was applied more than 3 hours after 2 Gy irradiation for NMT1-R. CONCLUSION: These results from our cell lines with the same origin were useful for investigation into the interaction of irradiation with heat. PMID- 12530058 TI - Down's syndrome-associated single minded gene as a novel tumor marker. AB - The Cancer Genome Anatomy Project (CGAP) database has thousands of Expressed Sequence Tags encompassing both known and novel genes. Bioinformatics of the CGAP database led to the prediction that Single Minded Gene (sim2) could be specific to colon tumors. The sim2 gene is located in a minimum region of the chromosome 21 often implicated in trisomia called Down's Syndrome Critical Region. To date, the sim proteins have not been shown to be involved in cancer. Intrigued by the possible association of a Down's syndrome-related gene to solid tumors, efforts were undertaken to validate the expression specificity. The sim2 isoform (sim2 short-form, sim2-s) expression was seen in carcinomas of colon, pancreas and prostate, but not in corresponding normal tissues. Stage-specific expression of the sim2-s protein was seen in normal matched paraffin sections of the colon tumors. In a matched set of tissues of Benign Prostatic Hyperplasia (BPH) and prostate carcinomas, sim2-s expression was detected in the BPH. The expression specificity of sim2-s in select solid tumors offers both diagnostic and therapeutic potential and warrants additional study. PMID- 12530059 TI - Modulation of cdk2, cyclin D1, p16INK4a, p21WAF and p27Kip1 expression in endothelial cells by TNF/IFN gamma. AB - BACKGROUND: Regional administration of high doses of tumor necrosis factor (TNF) and interferon gamma (IFN gamma) to metastatic melanoma patients causes selective disruption of the tumor vasculature. This effect is paralleled by decreased endothelial cell proliferation and suppressed integrin alpha V beta 3-mediated adhesion in vitro. Overexpression of the cyclin-dependent kinase (cdk) inhibitory protein p16INK4a was reported to interfere with integrin alpha V beta 3-dependent melanoma cell adhesion. MATERIALS AND METHODS: TNF- and IFN gamma-treated HUVEC were analyzed for cell cycle progression and for protein expression by flow cytometry and Western blotting, respectively. p16INK4a was overexpressed by transient transfection, and HUVEC adhesion was tested in short-term adhesion assays. RESULTS: TNF and IFN gamma synergistically induced a G1 arrest associated with reduced levels of cyclin D1 and cdk2, and increased expression of the cdk inhibitors p16INK4a, p21WAF and p27Kip1. p16INK4a overexpression, however, had no effect on alpha V beta 3-mediated adhesion. CONCLUSION: These results implicate the down-regulation of cyclin D1 and cdk-2, and up-regulation of p16INK4a, p21WAF and p27Kip1 in the suppression of endothelial cell proliferation induced by TNF/IFN gamma and demonstrate that increased p16INK4a levels are not sufficient to suppress alpha V beta 3-mediated endothelial cell adhesion. PMID- 12530060 TI - Overexpression of ICBP90, a novel CCAAT-binding protein, overcomes cell contact inhibition by forcing topoisomerase II alpha expression. AB - We have recently identified ICBP90 as being a protein able to bind in vitro a CCAAT box of the topoisomerase II alpha gene promoter. The aim of the present work was to check whether ICBP90 is able to regulate in vivo topoisomerase II alpha expression in human lung fibroblasts under various proliferating conditions. Transient transfection experiments performed on moderately growing human lung fibroblasts (50% of confluence) showed that overexpression of ICBP90 is associated with an elevation of topoisomerase II alpha expression and an increase of the cell proliferation rate. In highly proliferating human lung fibroblasts (20% confluence) overexpression of ICBP90 had no effect. In contrast, in non-proliferating fibroblasts (100% confluence) overexpression of ICBP90 allowed recovery of topoisomerase II alpha expression levels with a concomitant overgrowth of confluent cell cultures. Our results show that ICBP90 regulates topoisomerase II alpha expression and is able to overcome cell contact inhibition signaling, suggesting that increased ICBP90 expression may be involved in carcinogenesis. PMID- 12530061 TI - Characterization of phosphodiesterase 3 in human malignant melanoma cell line. AB - Little is known concerning the expression, distribution and function of phosphodiesterase (PDE) 3s in malignant tumor cells, including human malignant melanoma HMG and osteosarcoma HOSM-1 cells. PDE3 activity was detected in homogenates of HMG cells; however, much less activity was found in HOSM-1 cells. In HMG cells, most of the PDE3 activity was in the particulate fraction. PDE3A and 3B mRNAs were detected by RT-PCR in RNA from HMG cells only. The nucleotide sequences of the fragments were identical to those of human PDE3A and 3B. The PDE3-specific inhibitors, trequinsin and cilostamide, did not inhibit the proliferation of HMG or HOSM-1 cells. Although two PDE3 isoforms may be expressed in human malignant melanoma cells, their functional importance is not known. PMID- 12530062 TI - Sustained tyrosine-phosphorylation of FAK through Rho-dependent adhesion to fibronectin is essential for cancer cell migration. AB - BACKGROUND: Co-stimulation with lysophosphatidic acid (LPA) and fibronectin (FN) is essential for migration of rat ascites hepatoma MMI cells. We examined the roles of LPA and FN in Rho-FAK pathway to migration. MATERIALS AND METHODS: We evaluated the morphology, phagokinetic motility and the status of Rho activation and tyrosine-phosphorylation of FAK after stimulation of MMI or HT-1080 cells with LPA + FN or each alone. RESULTS: On FN-coated dishes without LPA, MM1 cells could not migrate and harbored undetectable levels of activated RhoA. Stimulation with LPA + FN enabled the MM1 cells to migrate and bear active RhoA and sustained tyrosine-phosphorylation of FAK. To the contrary, HT-1080 cells could migrate and harbored a significant amount of active RhoA accompanied by sustained tyrosine phosphorylation of FAK even without LPA. CONCLUSION: Rho-dependent adhesion to FN leading to sustained tyrosine-phosphorylation of FAK is essential for cancer cell migration. PMID- 12530063 TI - Efficient translocation and apoptosis induction by adenovirus encoded VP22-p53 fusion protein in human tumor cells in vitro. AB - BACKGROUND: p53 mutations are one of the most frequent genetic alterations in cancer. Various mechanisms of delivering p53 protein into tumor cells, such as plasmids, retroviruses and adenoviruses, have been widely used in experimental studies. Although these methods are relatively successful, the transduction rate into surrounding cells is still a limiting factor. Recent studies have shown that fusing VP22 (important for intercellular transport) to p53 and delivering this protein to cells in the form of an adenovirus is a very efficient method of getting p53 into cells. RESULTS: In the present study, the effect of adenovirus encoded VP22-p53 fusion protein on p53 negative human cancer cells (LNCaP, SK-OV 3, OVCAR-3, DU 145) was investigated. A functional VP22-p53 fusion protein was produced as verified by immunofluorescence and analysis of p21 expression. Induction of p21 expression (target gene for p53) confirmed p53 was functional and immunofluorescence staining using p53 antiserum demonstrated the intercellular trafficking ability of the VP22 portion of the fusion protein. CONCLUSION: In conclusion, VP22-p53 showed efficient translocation into tumor cells, inhibition of tumor cell proliferation and induction of apoptosis. These characteristics make the fusion protein an attractive method for introducing p53 into human cells as well as a potential candidate for gene therapy. PMID- 12530064 TI - Expression of p57kip2 and its clinical relevance in epithelial ovarian tumors. AB - BACKGROUND: The aim of this study was to elucidate the expression of p57kip2 in a series of benign, borderline and malignant ovarian tumors, to evaluate its relationship with other G1 regulators of the cell cycle and to determine whether p57kip2 expression is associated with progression and prognosis of epithelial ovarian tumors. MATERIALS AND METHODS: Immunohistochemical analysis was performed in 103 cases of epithelial ovarian tumors. Twenty-six of the 103 cases were evaluated by Western blot analysis. RESULTS: The study demonstrated that high p57kip2 expression was detected in 63.6% (21 out of 33) of benign tumors, 52.2% (12 out of 23) of borderline tumors and 40.4% (19 out of 47) of ovarian carcinomas. The positive ratio of p57kip2 expression was decreased from benign to borderline to malignant tumors, whilst statistical significance was observed between benign and malignant tumors. Low p57kip2 expression was significantly associated with high tumor grades, advanced clinical stages and cyclin E overexpression. Kaplan-Meier analysis demonstrated that the patients with low p57kip2 expression had a short overall survival. When the combined phenotype of p57kip2 and p27kip1 was analyzed, the patients with both p57kip2 and p27kip1 low expression had a lower overall survival rate. CONCLUSION: These findings suggest that decreased p57kip2 expression may play a pivotal role in the progression of ovarian tumors and provide an important prognostic implication for epithelial ovarian carcinomas. PMID- 12530066 TI - Loss of heterozygosity at the FHIT gene in different solid human tumours and its association with survival in colorectal cancer patients. AB - BACKGROUND: Genomic alterations and abnormal expression of the FHIT gene have been reported for a number of cancers. FHIT encompasses FRA3B, the most common fragile site in the human genome, and is suggested to be a candidate tumour suppressor gene. MATERIALS AND METHODS: We analysed and compared the loss of heterozygosity (LOH) pattern in 397 solid human tumours from 9 different locations, using four polymorphic microsatellite markers within the gene (D3S1234, D3S1300, D3S2757 and D3S4260), and two markers (D3S1313 and D3S1600) flanking the gene. In addition, we tested whether there was an association between FHIT LOH and overall patient survival in colorectal cancer. RESULTS: LOH at the FHIT gene affecting at least one of the investigated markers was detected in 166 out of 332 informative tumours, or 50%. The highest detected LOH was in lung tumours (66%) while the lowest was in thyroid and endometrium tumours, (30% and 31%, respectively). Breakpoints were found inside the gene in all tumour types in 12-80% of the tumours with FHIT LOH depending on tumour type, and up to 41% could additionally be located adjacent to the 3' or 5' end of the FHIT gene. Thus we were able to locate breakpoints within or in the vicinity of the FHIT gene in 25-100% of different tumours with LOH. Although not statistically significant, we observed a trend towards a poorer survival of patients with FHIT LOH versus those with retention of heterozygosity. CONCLUSION: Based on our results, LOH of the FHIT gene is a common event in all tumour types analysed with a possible association with poorer survival in colorectal cancer patients. LOH at all markers analysed was, in most of the tumour types, a more common pattern of alterations than breakpoints. PMID- 12530065 TI - Reduced expression of p53 and p21WAF1/CIP1 sensitizes human breast cancer cells to paclitaxel and its combination with 5-fluorouracil. AB - BACKGROUND: The favorable clinical activity of paclitaxel has prompted considerable interest in combining this agent with other clinically effective antineoplastic agents including the antimetabolite 5-fluorouracil (5-FU). Our previous studies indicated that simultaneous exposure or pretreatment with 5-FU could significantly interfere with the cytotoxic effects of paclitaxel on both mitotic arrest and apoptosis. Biochemical examination also revealed that 5-FU inhibited expression of p21WAF1/CIP1 that may contribute to paclitaxel cytotoxicity. MATERIALS AND METHODS: In this study, human breast cancer BCap37 cells were transfected with either sense or antisense p53 or p21WAF1/CIP1. The established stable transfectants were then analyzed for an altered sensitivity to paclitaxel, 5-FU or the combinations of these drugs using a series of cytotoxic and apoptosis assays. RESULTS: Tumor cells transfected with antisense p53 or p21WAF1/CIP1 exhibited a significant increase in their sensitivity to paclitaxel. The reduced protein levels of p53 and p21WAF1/CIP1 were also found to abrogate the inhibitory effects of 5-FU on paclitaxel-induced mitotic arrest and apoptosis. CONCLUSION: These findings suggest that the status of p53 and p21WAF1/CIP1 might play an important role in tumor cell susceptibility to paclitaxel and its combination with other drugs such as 5-FU. PMID- 12530067 TI - Identification of differentially expressed genes in classical and atypical multidrug-resistant gastric carcinoma cells. AB - The phenomenon of multidrug resistance (MDR) in human cancers is the major cause of failure of chemotherapy. To better understand the molecular events associated with the development of different types of MDR, a classical MDR P-glycoprotein expressing gastric carcinoma cell line and an atypical MDR P-glycoprotein negative variant were analyzed by cDNA array hybridization. Of 588 cDNAs spotted on the array, a total of 9 genes showed differences in mRNA expressions. An enhanced expression level of 3 genes could be detected in both different MDR models (HLH, IR21, CCT5, Tx P-1), 4 genes were overexpressed solely in classical MDR cells (Hsp27, Rcl, aldehyde dehydrogenase 1, vimentin), whereas the expression of one gene was increased in atypical MDR cells (ProTa). Furthermore, the mRNA expressions of 3 genes were down-regulated in atypical MDR cells (Hsp27, vimentin, JNK2), whereas a decrease in mRNA expression in classical MDR cells could be determined for none of them. These differences were confirmed by a semiquantitative RT-PCR approach. Further characterization of these factors may provide more insight into the biology and development of different types of MDR in human gastric carcinomas. Moreover, these genes may be potential candidate factors for the diagnostics and/or prognosis of clinical drug resistance. PMID- 12530068 TI - Intrinsic and extrinsic manipulation of B7/CTLA-4 interaction for induction of anti-tumor immunity against osteosarcoma cells. AB - BACKGROUND: B7 family members play a central costimulatory role in T cell activation. We identified B7-1a, an alternatively spliced form of B7-1. The therapeutic efficacy of B7-1a-expressing tumor vaccine remains uncertain. MATERIALS AND METHODS: The murine osteosarcoma cell line, LM8, was engineered to express equivalent levels of B7-1 (B7-1-LM8) and B7-1a (B7-1a-LM8). The therapeutic efficacy of B7-transfected cells and anti-CTLA-4 blocking mAb was evaluated by the mixing experiments on the primary tumor, pulmonary metastasis and survival time. RESULTS: (i) a mixture of B7-1-LM8 or B7-1a-LM8 cells inhibited growth of the subcutaneous LM8 tumors and augmented the therapeutic effects of anti-CTLA-4 mAb and (ii) a combination of B7-1a-LM8 cells and anti CTLA-4 mAb most significantly eradicated pulmonary metastasis and prolonged the survival time of mice. CONCLUSION: These findings suggest that intrinsic (lack of IgC-like domain in B7-1a) and extrinsic (anti-CTLA-4 mAb) manipulations of B7/CTLA-4 interaction synergistically improve the therapeutic efficacy of B7 based osteosarcoma vaccines. PMID- 12530069 TI - Enhancement of anti-tumor immunity by lipoteichoic acid-related molecule isolated from OK-432, a streptococcal agent, in athymic nude mice bearing human salivary adenocarcinoma: role of natural killer cells. AB - BACKGROUND: OK-PSA, a lipoteichoic acid (LTA)-related molecule isolated from a streptococcal agent OK-432, enhances anti-tumor immunity as a potent inducer of Th1-type cytokines. Recently, we obtained the data suggesting that natural killer (NK) cells may play a significant role for OK-PSA-induced cytokine production in vitro. MATERIALS AND METHODS: We conducted the animal experiments using athymic nude mice bearing human salivary adenocarcinoma to examine the role of NK cells in OK-PSA-induced anti-tumor immunity. OK-PSA was peritumorally injected into the mice. Cytokines in the sera were analyzed by ELISA. mRNAs for cytokines were detected by RT-PCR. 51Cr release test was performed to measure killer cell activities. RESULTS: OK-PSA markedly increased the amounts of IFN-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-2, IL-12 and IL-18 that are generally called "Th1-type cytokines" in the sera derived from tumor-bearing nude mice, and also accelerated the killing activities of tumor-infiltrating lymphocytes as well as of draining lymph node cells. Furthermore, OK-PSA administration resulted in significant inhibition of tumor growth, but the effect of OK-PSA was almost completely inhibited by the deletion of NK cells using anti asialo GM1 antibody. CONCLUSION: These findings strongly suggested that NK cells are closely involved in OK-PSA-mediated anti-tumor immunity. PMID- 12530070 TI - High Ki-67 labeling index in human colorectal cancer with microsatellite instability. AB - BACKGROUND: In the present study, we examined the relationships between microsatellite status and Ki-67 labeling index, a proliferative marker, in colorectal cancers in order to clarify the common biological profiles of familial and sporadic cases of MSI (microsatellite inability)-positive tumors. PATIENTS AND METHODS: Seventy-eight invasive colorectal carcinomas were studied. Five microsatellite loci were analyzed by polymerase chain reaction, while Ki-67 was studied by immunohistochemistry. RESULTS: The MSI was found in 15 (19.2%) of the 78 tumors, and the Ki-67 labeling index was significantly higher in the 15 MSI positive tumors than that in the 15 MSI-negative ones (p = 0.0181). Moreover, the Ki-67 labeling index was significantly higher in the 7 tumors with MSI at more than 2 loci (MSI-H; high-frequency MSI) than that in the MSI-negative ones (p = 0.0082). CONCLUSION: The present study demonstrates that colorectal cancers with MSI show high Ki-67 labeling index irrespective of hereditary or nonfamilial cancer types, suggesting their common biological aspects distinct from MSI negative ones. PMID- 12530071 TI - A potential important role for thymidylate synthetase inhibition on antitumor activity of fluoropyrimidine and raltitexed. AB - BACKGROUND: Because thymidylate synthetase (TS) is a key enzyme in DNA synthesis, it has been used as a target for cancer chemotherapy. MATERIALS AND METHODS: We investigated the combined antitumor activity of raltitexed, 5-FU and UFT on human tumor xenografts in nude mice and examined changes in TS activity and 5-FU-bound RNA (F-RNA) levels. Human gastric (SC-1-NU) or colon (HT-29) carcinoma xenografts were transplanted subcutaneously into nude mice, and drugs administered intraperitoneally (raltitexed and 5-FU) or perorally (UFT) daily for 5 days, and repeated once after a 2-day interval. RESULTS: The antitumor effects were mostly equivalent between the treatment groups despite the different drugs and sequence orders. TS inhibition rates correlated with the tumor inhibition rate, which was statistically significant, while F-RNA levels did not correlate with antitumor activity. CONCLUSION: Our results indicated that the combination of fluoropyrimidine-related agents should be directed towards increased TS inhibition rather than increased F-RNA levels. PMID- 12530072 TI - Augmented growth inhibition of B16-BL6 melanoma by combined treatment with a selective matrix metalloproteinase inhibitor, MMI-166, and cytotoxic agents. AB - BACKGROUND: MMI-166 is a selective matrix metalloproteinase (MMP) inhibitor. The purpose of this study was to evaluate the antitumor efficacy of the combined treatment of MMI-166 with paclitaxel or carboplatin. MATERIALS AND METHODS: Mice bearing B16-BL6 melanoma were treated p.o. with MMI-166 from 1 day after tumor inoculation. The mice were administered i.v. with either paclitaxel or carboplatin at the maximum tolerated dose (MTD). RESULTS: MMI-166 monotherapy inhibited in vivo growth of the B16-BL6 tumor to an extent similar to that of paclitaxel or carboplatin monotherapy. When MMI-166 was combined with paclitaxel or carboplatin, the antitumor efficacy was significantly (p < 0.01) augmented in comparison with either MMI-166 or each cytotoxic agent alone. The hematotoxicity study demonstrated that daily treatment with MMI-166 did not affect the blood cell number in the mice and more importantly the combination of MMI-166 with paclitaxel did not augment the hematotoxicity caused by paclitaxel. An in vitro cytotoxicity study showed that MMI-166 itself has neither direct cytotoxicity against B16-BL6 tumor cells nor does it augment the cytotoxicity of paclitaxel or carboplatin. CONCLUSION: These results indicate that augmented antitumor activity of the combination treatment was not simply due to the augmentation of direct cytotoxic activity, but was rather an additive effect of the antitumor activities of different mechanisms. They suggest the effectiveness of a combination therapy of MMI-166 with paclitaxel or carboplatin in clinical therapy. PMID- 12530073 TI - Optimization of tumor-targeted gene delivery by engineered attenuated Salmonella typhimurium. AB - BACKGROUND: Attenuated Salmonella typhimurium has been demonstrated as a potential gene delivery vector. Previous findings induce the necessity to optimize tumor selectivity and bacterial dosing in relation to tumor volume and intratumoral therapeutic gene expression. MATERIALS AND METHODS: Attenuated Salmonella VNP20009 and VNP20047 (expressing cytosine deaminase) were systemically administered to tumor-bearing rats. The bacteria were quantified in tumor and normal organs. Conversion of 5-fluorocytosine to 5-fluorouracil was evaluated using thin layer chromatography. RESULTS: Tumor colonization efficiency was dependent on Salmonella density, administration route and tumor volume. Colonization of normal tissues gradually decreased with time, while intratumoral proliferation of bacteria remained high during the follow-up period. The Optimal Therapeutic Dose (OTD) was found to be 5.10(7) cfu/rat. Intratumoral VNP20047 expressed CDase leading to the conversion of 5-FC to 5-FU was detected in vivo. CONCLUSION: Our results indicate the need to define an OTD, probably for each species, when using genetically engineered Salmonella as a tumor- and species selective vector in cancer therapy. PMID- 12530074 TI - Selective protection of non-cancer cells by hypothermia. AB - BACKGROUND: A serious limitation in cancer treatments is insufficient selectivity of drugs for cancer cells. We have previously demonstrated that, in contrast to p53-deficient cells, cells with wild-type p53 undergo a reversible cell cycle arrest when incubated at 28 degrees C instead of 37 degrees C. Since most of the human tumors are p53-deficient, it suggests that hypothermia may selectively protect normal cells from cytotoxic treatments that primarily target proliferating cells. MATERIALS AND METHODS: We have examined the effect of hypothermia on the survival of wild-type and p53-deficient cells exposed to the anti-tumor drug 5-fluorouracil and compared BrdU incorporation at 28 degrees C and 37 degrees C of normal and tumor cells. RESULTS: p53 wild-type fibroblasts, in contrast to p53-deficient cells, survive much higher doses of 5-fluorouracil when incubated at 28 degrees C than at 37 degrees C. Among tumor cells, the loss of the p53 function coincides with the inability to arrest cell cycle progression at low temperature and with increased sensitivity to prolonged hypothermia as a single modality. CONCLUSION: Hypothermia protects normal cells from cytotoxic treatments and may improve the therapeutic index of chemotherapy by mechanisms based on the differences in cell cycle regulation between normal and tumor cells. PMID- 12530075 TI - Lymphogenic metastatic spread of auricular VX2 carcinoma in New Zealand white rabbits. AB - INTRODUCTION: The auricular VX2 carcinoma of the New Zealand white rabbit serves as an animal model for human squamous cell carcinomas of the head and neck region (HNSCC), since both tumors tend to metastasize lymphatically, leading to early lymph node and subsequent distant metastasis. The aim of this study was to examine the pattern of lymphogenic metastatic spread in untreated auricular VX2 carcinomas, since the resulting knowledge potentially could help in the development of new treatment strategies for human HNSCC. MATERIALS AND METHODS: VX2 carcinomas were implanted into both ears of 22 New Zealand white rabbits. The animals were sacrificed at days 7, 14, 21, 28 or 32 after tumor implantation, followed by a detailed histopathological examination of their head and neck lymph nodes. RESULTS: On day 7 after tumor implantation 25% of the animals had metastases in the parotid lymph node, which is the first draining lymph node of the tumor region. This number rose to 87.5% by day 28. At this time 12.5% of all animals also had an additional metastasis in the second echelon node. CONCLUSION: A reproducible metastatic spread into the first draining lymph node could be demonstrated for the auricular VX2 carcinoma of the New Zealand white rabbit. The VX2 carcinoma therefore appears to be a highly suitable animal model for studying the sentinel node concept in the context of human HNSCC. PMID- 12530077 TI - Amplified Muc1-specific gene expression in colon cancer cells utilizing a binary system in adenoviral vectors. AB - Mucin-1 is expressed in a variety of colon carcinomas and Muc-1/DF3 promoters have been utilized to reduce systemic toxicity through specific gene expression. To overcome weak expression, which is much lower than the widely used cytomegalovirus-promoter (CMV), new adenoviral vectors containing a binary system of transgene amplification have been developed. The Muc-1/DF3 promoter was used to control the expression of a Gal4VP16 fusion protein. This vector also contained Gal4 binding sites enabling the fusion protein to act as a transactivator, inducing transgene expression within the same construct. Mucin-1 expression was analyzed in a variety of colon cancer cell lines. After infection with recombinant adenoviruses, transgene expression was quantified using the luciferase system. Integration of the Gal4VP16-binary resulted in an up to 250 fold increase of Muc-1/DF3-specific gene expression. In mucin-positive cell lines utilizing this amplified Muc-1/DF3 promoter, expression was up to 590-fold higher as compared to the CMV-promoter. Western blot detected the presence of Gal4VP16 in infected muc-1-positive but not-negative cell lines. These new adenoviral vectors combing highly efficient and specific transgene expression and will contribute to the safety and efficacy of experimental approaches in cancer gene therapy. PMID- 12530076 TI - Expression of MMP-3, MMP-13, TIMP-2 and TIMP-3 in the VX2 carcinoma of the New Zealand white rabbit. AB - BACKGROUND: Squamous cell carcinomas of the head and neck region (HNSCC) are among the most common malignancies in this area. The VX2 carcinoma of the New Zealand white rabbit metastasizes lymphatically as is the case in HNSCC and therefore, potentially, could be used as a model for HNSCC. Since the family of matrix-metalloproteinases (MMPs) is involved in the process of HNSCC invasion, the aim of this study was to investigate the expression level of MMPs and their specific inhibitors (TIMPs) in the VX2 carcinoma to evaluate if they also play a role in VX2 tumor invasion as observed in human HNSCC. MATERIALS AND METHODS: The VX2 carcinoma was generated by tumor implantation in the rabbit's ear as previously described. Western blots were performed under standard conditions, utilizing antibodies against MMP-3, MMP-13, TIMP-2 and TIMP-3. Immunohistochemical staining was performed with the ABC-complex method. RESULTS: A positive immunohistochemical signal could be detected for MMP-3, TIMP-2 and TIMP-3 with no significant signal for MMP-13. In the Western blots immunoreactive bands could be observed for MMP-3, MMP-13, TIMP-2 and TIMP-3. CONCLUSION: MMP-3, MMP-13, TIMP-2 and TIMP-3 were found to be expressed in VX2 carcinomas of the New Zealand white rabbits. The VX2 carcinoma therefore resembles HNSCC tumors not only in its metastatic behavior, but also regarding the expression of MMPs and TIMPs, which are the probable keyplayers during the event of invasion. These observations further underline the significance of the VX2 carcinoma as a model tumor of human HNSCC. PMID- 12530079 TI - Down-regulation of TRF1, TRF2 and TIN2 genes is important to maintain telomeric DNA for gastric cancers. AB - BACKGROUND: The maintenance of telomeres may be required for long-term proliferation of tumors. Activity of telomerase, a ribonucleoprotein complex that elongates telomeres, has been found in almost all human tumors but not in adjacent normal cells. Several factors which regulate telomere length, TRF1 and 2, TIN2, tankyrase and Rap1, have been identified. TRF1, TRF2 and TIN2 are negative regulators of telomere length, while tankyrase and Rap1 act as positive regulators. In this study, we quantitated the mRNA of these five genes in gastric cancers to clarify the mechanism by which cancer cells maintain telomere length. MATERIALS AND METHODS: The expression of these five genes transcription was determined using a quantitative RT-PCR. RESULTS: TRF1, TRF2 and TIN2 mRNAs were significantly down-regulated in cancers compared to non-cancerous mucosa. Neither tankyrase nor Rap1 was upregulated in cancers. CONCLUSION: Down-regulation of TRF1, TRF2 and TIN2 gene expression may be important to maintain telomeres in gastric cancer. PMID- 12530078 TI - Effect of poly-herbal formula on NO production by LPS-stimulated mouse macrophage like cells. AB - Pretreatment of mice with lyophilized hot water extracts of five poly-herbal formula protected them from lethal infection by E. coli. ESR spectroscopy shows that these extracts produced radicals under alkaline condition, and scavenged radicals such as superoxide anion, hydroxyl radical and nitric oxide (NO) radical. There was a positive relationship between their radical intensity and radical scavenging activity. Among the extracts, HD-02 efficiently inhibited the production of NO and citrulline, and the expression of inducible NO synthase (iNOS) mRNA by LPS-stimulated mouse macrophage-like cells Raw 264.7. DLH-3073 not only inhibited the LPS-stimulated NO production at lower concentration, but also induced NO production at higher concentrations, suggesting the presence of two different antagonizing components in the DLH-3073 extract. These data suggest that poly-herbal extracts may alleviate radical-mediated diseases. PMID- 12530080 TI - Antitumor and antimetastatic effects on liver of triterpenoid fractions of Ganoderma lucidum: mechanism of action and isolation of an active substance. AB - The triterpenoid fraction (100 and 200 mg/kg) of the fruit bodies of Ganoderma lucidum inhibited primary solid-tumor growth in the spleen, liver metastasis and secondary metastatic tumor growth in the liver in intrasplenic Lewis lung carcinoma (LLC)-implanted mice. In addition, the triterpenoid fraction (800 micrograms/mL) inhibited angiogenesis induced by Matrigel (a soluble basement membrane extract of the Engelbreth-Holm-Swam (EHS) tumor) supplemented with vascular endothelial growth factor (VEGF) and heparin in an in vivo model. This suggested that the antitumor and antimetastatic activities of the triterpenoid fraction of G. lucidum might be due to the inhibition of tumor-induced angiogenesis. Next, we attempted to isolate the active substance(s) using the in vivo assay system of Matrigel-induced angiogenesis. The acidic fraction of the triterpenoid fraction inhibited the Matrigel-induced angiogenesis. Compound I was isolated from the acidic fraction as an active substance that inhibited the Martigel-induced angiogenesis. Compound I was identified as ganoderic acid F based on the data of IR, 1H- and 13C-NMR and MS analyses. PMID- 12530081 TI - Non-toxic potentiation of cancer radiotherapy by dietary oligofructose or inulin. AB - Non-toxic, dietary treatment with oligofructose or inulin clearly inhibited the growth of a transplantable mouse liver tumor (TLT) and potentiated its chemotherapy. Thus, it appeared interesting to investigate the possible radiotherapy-potentiating effects of the same dietary treatment. Dietary treatment with 15% oligofructose or inulin incorporated in the basal diet was started four weeks before intramuscular transplantation of TLT tumor cells into young adult male mice of the NMRI strain and was continued until the end of the experiment. When the tumors reached approximately 1000 mm3 they were irradiated with a single X-ray dose of 5 to 20 Gy. Tumor dimensions were measured twice weekly and their mean volume per group of mice was compared to the control groups fed the basal diet. This non-toxic dietary treatment with oligofructose or inulin potentiated the effects of radiotherapy at an optimal dose of 10 Gy to a statistically very highly significant (p < 0.0001) level. They were similar for oligofructose and inulin. The introduction of such non-toxic adjuvant treatment potentiating the effect of cancer radiotherapy in classical protocols of human cancer treatment appears to be possible and without any additional risk for the patients. PMID- 12530082 TI - Comparison of the EF-1 alpha and the CMV promoter for engineering stable tumor cell lines using recombinant adeno-associated virus. AB - BACKGROUND: Silencing of the viral CMV immediate early enhancer promoter can be a problem in certain cell types when engineering stable cell lines. MATERIALS AND METHODS: We compared the efficacy of the CMV promoter to the promoter of the elongation factor-1 alpha (EF-1 alpha) for the generation of stable colon carcinoma cell lines (HT-29). Green fluorescent protein (GFP) expression cassettes were delivered by recombinant adeno-associated virus (AAV) which is known for its ability to stably transduce cells. Stable cell lines were characterized in vitro by FACS and in vivo after HT-29 clones were grown as xenografts in nude mice. RESULTS: Stable HT-29 clones with > 97% of all cells homogeneously expressing GFP were generated with the EF-1 alpha promoter. In contrast in clones carrying the CMV promoter, only up to 60% of the cells were GFP-positive with expression levels varying widely between cells. Superinfection with wild-type adenovirus induced GFP expression in more than 90% of the cells indicating that the CMV promoter was silenced. In vivo the tumors carrying the EF 1 alpha promoter were homogeneously GFP-positive, whereas the CMV promoter gave rise to a scattered pattern of GFP expression. CONCLUSION: This study underlines the importance of the promoter for the generation of stable cell lines. In addition it demonstrates that recombinant AAV can effectively be used as a gene delivery system for this purpose. PMID- 12530083 TI - Overexpression of mitochondrial thioredoxin reductase and peroxiredoxin III in hepatocellular carcinomas. AB - Thioredoxin reductase 2 (TrxR2), thioredoxin II (Trx II) and peroxiredoxin III (Prx III) are specifically localized in mitochondria and believed to play important roles in the regulation of cellular redox status by serving as a primary line of defense against H2O2 produced during respiration. Substantial evidence indicates that the alteration of cellular redox status is a critical factor involved in cell growth and death and results in tumorigenesis. We therefore investigated the expression of TrxR2 and Prx III in 58 paraffin embedded hepatocellular carcinoma tissues by immunohistochemistry. The labeling indices of TrxR2 and Prx III were significantly higher in tumor tissues than in the corresponding adjacent normal tissues. In 39 (67.2%) out of 58 samples, the levels of TrxR2 expression were higher in tumor tissues than in corresponding adjacent normal tissues, while 11 samples (19.0%) showed lower expression in tumor tissues. Prx III expression was increased in tumor tissues of 23 samples (39.7%) compared to adjacent normal tissues and were decreased in 18 samples (31.0%). These results suggest that alterations in cellular redox status by enhanced expression of TrxR2 and/or Prx III might be associated with the formation and development of hepatocellular carcinomas. PMID- 12530084 TI - Expression of the KAI1 metastasis suppressor gene in non-metastatic versus metastatic human colorectal cancer. AB - BACKGROUND: The importance of KAI1 to colorectal cancer (CRC) progression is unclear, with conflicting data regarding changes in KAI1 expression during tumour progression. MATERIALS AND METHODS: In situ hybridisation and immunohistochemistry, with a semiquantitative scoring system, were used to assess KAI1 mRNA and protein levels, respectively, in normal colon samples (43), and non metastatic (24) or metastatic (33) primary cancers, and liver metastases (48). RESULTS: There was significant loss of KAI1 mRNA and protein staining in non metastatic primary tumours versus normal tissues (Bonferroni, p < 0.05) but levels recovered to near normal in metastatic primary tumours and liver metastases. Increased KAI1 expression significantly correlated with distant metastases (Mann-Whitney, p = 0.0001, mRNA; p = 0.033, protein), cancer-specific survival (Cox regression analysis p = 0.0002, mRNA; 0.0493, protein) and overall patient survival (p = 0.0001, mRNA). Multivariate analysis (Cox proportional hazards model) confirmed high KAI1 mRNA expression was an independent prognostic indicator of distant metastasis (p < 0.0001), cancer-specific survival (p < 0.0001) and overall patient survival (p < 0.0001). CONCLUSION: These data indicate a complex relationship between KAI1 and progression of human CRC, in which expression is reduced in localised primary tumours, but regained in disease associated with metastasis. PMID- 12530085 TI - Overexpression of the human myosin-binding protein-C1 mRNA in laryngeal squamous cell carcinoma cells. AB - BACKGROUND: To compare gene expression patterns between laryngeal squamous cell carcinoma (SCC) cells and their normal phenotypes to identify genes showing differential expression. MATERIALS AND METHODS: Messenger RNA was isolated from both kinds of cells, reversely transcribed and subjected to differential display reverse transcription (DDRT)-PCR. Gene fragments showing difference in the expression were recovered, reamplified, cloned and sequenced, enabling homology search. Total RNA was isolated from laryngeal SCC cells and adjacent normal mucosa and subjected to Northern hybridization. RESULTS: A 159 bp gene fragment was detected, revealing 96% homology with the human myosin-binding protein-C1 (MYBPC-1) gene. Compared to the benign phenotypes the expression of MYBPC-1 was particularly increased in SCC cells, confirmed by Northern hybridization. CONCLUSION: The results presented in this work may help to extend the diagnostic panoply available for the evaluation of laryngeal tissue conspicuous for malignancy. PMID- 12530086 TI - Early detection of chemoresistance in vivo through the use of a radiolabeled antisense oligonucleotide. AB - AIMS: Radiolabeled antisense oligonucleotide to target the mRNA of the hmdr1 gene for diagnostic purposes is a new concept for evaluating the chemoresistance of tumors in vivo. METHODS AND RESULTS: An 18 mer complementary to the zone which contains the translation initiation codon of the hmdr1 gene was modified using one phosphoramidate group and one dimethoxytrityle group at the 5' and 3'ends. It permitted probe radiolabeling by 125I. Chemical modifications made to the antisense probe ensured the stability in biological media tested by incubation with human serum at 37 degrees C from 5 minutes to 24 hours. These modifications did not interfere with recognition of the target. Retention of the antisense probe followed the expression level of the target transcript in in vitro and in vivo studies. In vitro, after a 2-hour incubation in the presence of K562- sensitive (S) and- resistant (R) cell lines, uptake was respectively 4.27 +/- 0.96% ID/mg protein and 7.78 +/- 0.46% ID/mg protein (p < 0.001). In vivo, the ratios between radioactivity found in the tumor and that found in the striated muscle and in the blood were, respectively, 20 and 3 for IGR OV1 resistant tumor and 1 and 0.3 for the sensitive one. CONCLUSION: In our study, resistant cell lines and tumor showed greater retention of the specific probe than the sensitive ones. This constitutes a further advance towards non invasive imaging of resistant genes involved in chemoresistance. These results are encouraging: the current trend in innovative cancer therapy is moving towards targeting the genes of interest. PMID- 12530087 TI - Comparison of various sources of antigen-presenting cells for the generation of GP2-tumor peptide specific cytotoxic T-lymphocytes. AB - BACKGROUND: The need for new therapeutic regimens for cancer has led to the identification of tumor antigens in several tumors. Yet, potent antigen presenting cells (APC) are being identified and characterized in order to find those most able to induce T-cell-mediated immunity. To facilitate the generation of cytotoxic T-lymphocytes (CTL) specific for known antigens, various sources of antigen-presenting cells (APC) were compared for their efficiency. MATERIALS AND METHODS: Allogeneic dendritic cells (DC), T2 cells and two Epstein Barr virus transformed B cell lines, JY and TK-6, were used as antigen-presenting cells (APC). T-cells from normal donors were stimulated with solid-phase anti-CD3 antibody and reactivated with peptide-pulsed APC three times at weekly intervals. To minimize any potential alloreactivity, eighteen random combinations of JY, TK6 and T2 were used for three peptide stimulations. The APC were pulsed with the HLA A2-binding HER2/neu-derived tumor antigen p654-662 (GP2). RESULTS: In addition to either T2 or DC alone, most combinations of APC induced significant recognition of GP-2-pulsed target cells to a varying degree. The stimulation index varied from 0.5 to 8.1 using combinations of APC. The level of peptide recognition correlated with recognition of HLA-A2+, HER2/neu+ pancreatic cancer cells; in contrast, HLA-A2-negative pancreatic cancer cells were not recognized. The recognition of tumor cells was HLA-restricted as evidenced by blocking studies with anti-HLA class I and anti-HLA-A2 antibodies. CONCLUSION: Tumor peptide specific CTL can be generated using allogeneic B cell lines, which may provide a useful alternative to T2 or dendritic cells. PMID- 12530088 TI - In vitro and in vivo pharmacological characterisation of the antitumour properties of pyrido[1',2':1,2]imidazo[4,5-h]quinazoline. AB - The anti-tumour activity of pyrido[1',2':1,2]imidazo[4,5-h]quinazoline (PIQ) was investigated in vitro and in vivo with a human tumour model. In vitro PIQ cytotoxicity was evaluated on two different human parental-sensitive cancer cell lines (HL60S and A2780S) and their multidrug-resistant variant sublines (HL60R and A2780R). Proliferation was assessed using the MTT assay and PIQ showed activity, particularly with resistant cell lines. Drug activity was not affected by MDR resistance. After LD50 determination using Swiss mice, in vivo activity with A2780 ovarian carcinoma was carried out using xenografted Swiss nude mice. We performed either a weekly intra-peritoneal injection of 64 mg.kg-1 PIQ or an intra-venous injection of 10 mg.kg-1 PIQ during 2 months. After 60 days of treatment, no toxicologically meaningful differences were observed in macroscopic and microscopic parameters compared to controls. Both regimens demonstrated efficacy against xenografted tumours. However, the decrease in tumoural volume of the xenografted mice was significant only in the PIC i.v. injection group. Pharmacokinetics and the accumulation of PIQ in normal and tumour tissues were also assessed using a chromatographic method. The lack of activity using the i.p. route was explained by the four-fold reduction of its AUC in comparison to the i.v. route. After an i.v. injection, the highest concentrations of PIQ were accumulated in the tumour and spleen. Drug analysis has shown that PIQ intercalates into DNA. PIQ derivatives are effective new antitumour agents in cancer chemotherapy. PMID- 12530089 TI - Induction of apoptosis by the green tea flavonol (-)-epigallocatechin-3-gallate in human endothelial ECV 304 cells. AB - We have previously shown that treatment with (-)-epigallocatechin-3-gallate (EGCG) inhibited vascularity and tumor growth in human colon cancer xenografts in nude mice (Jung et al: Br J Cancer 84, 2001). In this study, we examined whether endothelial cell death by EGCG is mediated by apoptosis and which molecular mechanisms are involved in this process. EGCG was found to suppress cell growth and induce apoptosis largely through mitochondrial depolarization, activation of caspase-3 and cleavage of DNA fragmentation factor-45 in human endothelial ECV 304 cells. The induction of apoptosis by EGCG was confirmed by cleaved and condensed nuclear chromatin and DNA hypoploidy. These results suggest that EGCG may exert at least part of its anticancer effect by inhibiting angiogenesis through inducing endothelial apoptosis. PMID- 12530090 TI - TP53 codon 72 polymorphism in oral squamous cell carcinoma. AB - BACKGROUND: Oral squamous cell carcinoma (OSCC) is a multifactorial neoplasm related to tobacco smoke, alcohol consumption and molecular genetic changes such as p53 mutations. The human tumor suppressor gene TP53 contains single nucleotide polymorphism that encodes either arginin (Arg) or proline (Pro) at amino acid codon 72 of the p53 protein. The relationship between human cancer susceptibility and p53 polymorphism at codon 72 is controversial. The aim of this study was to investigate the association between this polymorphism and OSCC development in a Brazilian population. MATERIALS AND METHODS: Eighty-two patients with OSCC and 82 age-sex-matched controls were included in the study. DNA was extracted from all subjects' normal oral mucosa and the polymerase chain reaction amplification (PCR) was performed for detection of the TP53 genotypes. PCR products were analyzed in a 6.5% polycrylamide gel and silver-stained. Statistical analyses were performed with the chi 2 test and Fisher's exact test. RESULTS: The frequencies of the genotypes Arg/Arg, Arg/Pro and Pro/Pro, were 37.8%, 54.8% and 7.4%, respectively, for OSCC cases and 40.2%, 54.8% and 4.9% for controls. No significant differences in the distribution of TP53 genotypes were seen between the groups (p = 0.794). CONCLUSION: The present study does not support the hypothesis that this TP53 polymorphism is associated with oral cancer susceptibility. PMID- 12530091 TI - Expression of cathepsin D in urothelial carcinoma of the urinary bladder: an immunohistochemical study including correlations with extracellular matrix components, CD44, p53, Rb, c-erbB-2 and the proliferation indices. AB - The immunohistochemical Cathepsin D (CD) expression of tumor and stromal cells was investigated in a series of 77 urothelial carcinomas of the urinary bladder with the intention to evaluate its prognostic significance and its contribution to the metastatic potential of bladder cancer. CD expression (clone D13A) was correlated with the expression of extracellular matrix components (collagen type IV, laminin, fibronectin), CD44, p53, pRb, proliferation indices (PCNA and MIB1) as well as with other conventional clininopathological features. CD expression (> 10% of positive tumor cells) was observed in 77.9% of the carcinomas. Stromal CD expression was detected in all cases. Linear collagen type IV and laminin deposit at the tumor-stroma border (in > 25% of the BM) was found in 26% and 57.6% of the cases, respectively. The CD of cancer cells (CCCD) was inversely-correlated with the CD of the stromal cells (p = 0.039), tumor grade (p = 0.0028), tumor stage (p = 0.0046), p53 protein (p = 0.05) and positively-correlated with CD44 (p = 0.002) and pRb (p = 0.05). The stromal cells CD (SCCD) showed a statistically significant positive correlation with tumor grade (p < 0.0001) and stage (p = 0.0001), and the proliferation indices PCNA and MIB1 (p = 0.0001 and p = 0.0002, respectively). These data suggest that both CD of tumor and stromal cells could play important roles in the expansion of urothelial carcinoma of the urinary bladder. PMID- 12530092 TI - Modulation of ornithine decarboxylase activity and mitogen-activated protein kinases in protocatechualdehyde-induced apoptosis of CTLL-2 cells. AB - Protocatechualdehyde (PA, a dihydroxybenzene derivative) has previously been shown to induce apoptotic cell death in cytotoxic T cells (CTLL-2). However, the molecular mechanisms by which PA regulates apoptosis are still unclear. In this study, the possible roles of ornithine decarboxylase activity (ODC) and mitogen activated protein kinases (MAPKs) in the PA-induced apoptosis process were further investigated. We demonstrated that PA inhibited ODC activity induced by IL2 in a time- and dose-dependent manner. Furthermore, the expression of ODC mRNA stimulated by IL2 was also effectively suppressed. 0.12 mM PA inhibited the activation of ERK1/2 induced by IL2 and enhanced the activation of JNK, which was abrogated by IL2. No alteration in the effect of p38 MAPK on the apoptosis process was observed in the CTLL-2 cells. PD98059 (a specific ERK1/2 inhibitor) inhibited cell growth, led to cell apoptotic death and effectively decreased ODC activity and suppressed ERK1/2 activation induced by IL2. These data indicate that PA induced apoptosis in CTLL-2 cells by two mechanisms; either via inhibiting ODC induction or interfering with MAPK signaling pathways. PMID- 12530093 TI - The role of interleukin-8 in the initiation and progression of human cutaneous melanoma. AB - BACKGROUND: The expression of Interleukin-8 (IL-8) by human melanoma cells correlates with their metastatic potential in vitro and in nude mice. The role of IL-8 in human patients is not yet clear. The purpose of the present study was to search for a possible correlation between IL-8 in patients' melanoma specimens and the clinical course of the disease. PATIENTS AND METHODS: All patients operated on for malignant melanoma (MM) at our center between 1985 and 1993 were screened for the study. Only those with retrievable pathological and clinical material were included. IL-8 tumor levels were tested by immunohistochemistry. RESULTS: Seventy-four patients were eligible for inclusion in the study. In 49, the primary tumor was analyzed and in 25 a metastasis. IL-8 was expressed in 91% of the primary lesions, 70% of the lymph node metastasis and 70% in local recurrence or in-transit metastases. There was no correlation between level of expression and outcome. Comparing IL-8 expression in primary and secondary lesions of the same patient showed a trend towards better survival for those who had down-expression of IL-8. CONCLUSION: IL-8 is over-expressed in cutaneous MM. Although no direct correlation was found in the present study between IL-8 expression and survival, it would appear that IL-8 plays a major role in melanoma development. PMID- 12530095 TI - DNMT1 gene copy number does not influence susceptibility to development of malignant lymphoproliferative disease. AB - BACKGROUND: For many years, the methylating enzyme DNMT1 has been thought to critically influence neoplastic transformation. To determine if DNMT1 is involved in the early events of malignant transformation of lymphoid cells, we investigated whether allele-specific variation in the gene copy number of DNMT1 influences susceptibility to the development of malignant lymphoproliferative disease (LPD) associated with DNA hypermethylation. MATERIALS AND METHODS: DNMT1 gene copy number was assessed by subjecting DNA from DNMT1A/DNMT1B heterozygous patients suffering from benign LPD or malignant LPD featuring Myf-3 hypermethylation to Southern blotting and densitometric analysis. RESULTS: The distribution of DNMT1B to DNMT1A intensity ratios did not significantly differ between patients suffering from benign and patients suffering from malignant LPD and there was no association between increasing Myf-3 hypermethylation and increased DNMT1 gene copy number. CONCLUSION: In contrast to findings in mice, the results of this study exclude a role for DNMT1 gene copy number in susceptibility to neoplastic transformation in humans. PMID- 12530094 TI - Glutathione S-transferase (GSTM1 and GSTT1)-dependent risk for colorectal cancer. AB - BACKGROUND: The deleted GSTT1 and GSTM1 genotypes (null genotypes) resulting in loss of transferase activity are found in 10-20% and 50-60% of the population, respectively. PATIENTS AND METHODS: The GSTT1- and GSTM1-dependent risk for sporadic colorectal cancer (CRC) was studied in 247 incident CRC cases and 296 hospital-based controls. RESULTS: The GSTT1-null genotype was found to be 1.5 times more prevalent in CRC patients (17.4%) compared with controls (11.1%) (crude OR 1.6; p = 0.03). The GSTM1-null genotype was found to be equally prevalent in cases and controls (53%). Multivariate analysis showed a significant 1.7-fold risk for CRC associated with the GSTT1-null genotypes (p = 0.04) and this increased to 2.9 for smokers (p = 0.02). CONCLUSION: This study provides evidence of gene-environment interaction and illustrates the importance of further research into the role of genetic susceptibility for CRC. PMID- 12530096 TI - Overexpression of Wnt target genes in adenomas of familial adenomatous polyposis patients. AB - Germline mutations within the adenomatous polyposis coli (APC) gene, a key member of the Wnt signalling pathway, have been shown to cause adenoma development in familial adenomatous polyposis (FAP), a dominantly inherited predisposition to colorectal cancer. Although it has been suggested for several years that alterations within the Wnt pathway are the underlying events for the development of colorectal adenomas in FAP patients, no detailed analysis of the gene expressions of Wnt pathway members has been available in fresh colorectal tissue of FAP patients, so far. Thus, we investigated potential differences in the expressions of APC and its Wnt partners conductin, beta-catenin, cyclin D1, and c myc in normal colorectal mucosa and matched adenoma tissue of 14 FAP patients using real-time quantitative PCR. The expressions of both Wnt target genes, cyclin D1 and c-myc, were significantly increased in adenoma compared to matched normal mucosa. Furthermore, the overexpressions of these two genes showed a highly significant positive correlation. Our data suggest that the concomitant overexpression of the Wnt targets and cell cycle regulators cyclin D1 and c-myc plays an important role in the neoplastic proliferation of adenomas in FAP patients. PMID- 12530097 TI - The prognostic significance of epithelial-mesenchymal transition in breast cancer. AB - BACKGROUND: This study investigates the influence of epithelial-mesenchymal transition on prognosis in breast cancer. MATERIALS AND METHODS: Eighty breast carcinomas as well as 6 breast cancer cell lines were analysed immunohistochemically for the expression of epithelial keratins (K) K8, K19 and mesenchymal vimentin. Protein expression was correlated with histopathological factors and clinical follow-up. RESULTS: Suppression of K8 and K19 occurred in the majority of the tumours (72.5% and 65%), while aberrant expression of vimentin was found in 21.2% of the tumours. Suppression of K8 as well as expression of vimentin was significantly correlated with short survival (p < 0.004 and p < 0.006). Moreover HER2 overexpression significantly correlated with K19 (p < 0.0004) and vimentin (p < 0.0005). In cell lines with increasing invasive potential, epithelial keratins were lost in favour of mesenchymal vimentin. CONCLUSION: The transition from epithelial keratin to mesenchymal vimentin expression marks an important step in the malignant progression of breast cancer. PMID- 12530098 TI - Polymorphism of IL-1 receptor antagonist gene: role in cancer. AB - BACKGROUND: IL-1-cytokines are strongly involved in carcinogenesis and tumor genesis. In this systematic review we will describe the role of IL-1 RA within the cytokine network and its predictive and prognostic value in cancer. MATERIALS AND METHODS: We analysed preclinical and clinical studies. RESULTS: A polymorphism is present in intron 2 of the human IL-1RA gene. The IL-1RA allele 2 has an influence on the IL-1RA plasma level. The IL-1 beta level is elevated at the homozygous for IL-1RA, allele 2 (IL-1RN*2). The IL-1 gene cluster polymorphism is associated with gastric cancer. Carriers of IL-1 1B- 511T and IL 1RN*2 homozygotes have an increased risk of developing gastric cancer. Patients with a heterozygosity of allele 2 for IL-1 RA (IL-RA1/2) have an increased risk of cervical cancer. There are negative associations between IL-1 RA polymorphism and the occurrence of ovarian cancer. CONCLUSION: The polymorphism of IL-1RA seems to be involved in the induction of different solid tumors. PMID- 12530099 TI - PSC 833 modulation of multidrug resistance to paclitaxel in cultured human ovarian carcinoma cells leads to apoptosis. AB - BACKGROUND: Multidrug resistance (MDR) modulator PSC 833 has been shown to modulate multidrug resistance in Pg-p-positive human ovarian carcinoma cells A2780/ADR. Co-treatment of A2780/ADR cells with paclitaxel (PTX) and PSC 833 resulted in the restoration of PTX-sensitivity comparable to that in parental A2780 cells. RESULTS: The flow cytometry experiments presented here showed PTX (A2780) and PTX plus PSC 833 (A2780/ADR)-induced cell accumulation in the G2/M phase of the cell cycle with concomitant appearance of apoptotic cells with sub G0 (hypodiploid) DNA content. Furthermore, these events were accompanied by the appearance of poly(ADP-ribose) polymerase (PARP) cleavage, up-regulation of Bax, p53 and p21WAF1/CIP1 proteins and internucleosomal DNA fragmentation. Interestingly, we did not detect any significant alterations in Bcl-xL, CD95/Fas and Fas-L protein levels. CONCLUSION: These results demonstrate the PSC 833 reduced the Pg-p-mediated multidrug resistance in human ovarian carcinoma cells to PTX-induced apoptosis in vitro. PMID- 12530100 TI - Preterm premature rupture of the membranes and antioxidants: the free radical connection. AB - AIM: To discuss the role of oxidant stress in preterm, premature rupture of the membranes (PPROM). RESULTS: There is evidence to suggest that preterm, premature rupture of the membranes occurs secondary to focal collagen damage in the fetal membranes. CONCLUSION: Oxidant stress caused by increased ROS formation and/or antioxidant depletion may disrupt collagen and cause premature membrane rupture. We propose that supplementation with vitamins C and E may synergistically protect the fetal membranes, and decrease the risks of PPROM. PMID- 12530101 TI - Vaginal lactobacilli and preterm birth. AB - OBJECTIVE: To assess the relationship between the absence of vaginal lactobacilli and preterm birth at < 33 weeks of gestation. METHODS: A prospective study of the vaginal flora in the second trimester was undertaken in 1958 women with singleton pregnancies. The contribution of various microorganisms to preterm delivery was analyzed using a multivariate-logistic regression model. RESULTS: Lactobacillus species were not cultured from 28% of 118 women who delivered at < 33 weeks, 10% of 224 women who delivered between 33 and 36 weeks, and 5% of 1616 women who delivered at > 37 weeks of gestation. Lactobacilli (odds ratio and 95% confidence interval: 0.15 [0.09 to 0.24]), Mycoplasma hominis (2.3 [1.0 to 5.4]), and glucose non-fermentative gram-negative rods (2.1 [1.0 to 4.2]) were identified as independent risk factors for preterm delivery at < 33 weeks of gestation. Absence of lactobacilli (sensitivity and positive predictive value: 28% and 25%) was a better predictor of preterm delivery at < 33 weeks of gestation than the presence of Mycoplasma hominis (7% and 13%, respectively) or glucose non-fermentative rods (9% and 11%). CONCLUSIONS: Although this was not a cohort study, results suggest that tests for determining the presence of vaginal lactobacilli may be clinically useful tools for identifying women at an increased risk of preterm delivery at < 33 weeks of gestation. PMID- 12530102 TI - Socioeconomic and environmental risk factors of bacterial vaginosis in early pregnancy. AB - The main aim of this prospective study was to determine the socioeconomic, demographic and environmental factors that may be associated with the occurrence of bacterial vaginosis at early pregnancy in an indigent population from Central Poland. A group of 196 pregnant women was selected randomly from the patients of 10 district maternity units in the Lodz region, Central Poland. Only singleton pregnancies between 8 and 16 week of gestation were qualified for inclusion in the survey. A standard questionnaire covering medical, socioeconomic, demographic, constitutional and environmental items was administered to every subject and was verified with medical records. Cervico-vaginal swabs were collected from the women under study and tested for bacterial vaginosis (BV) according to Spiegel's criteria. Based on the results of Gram stain, BV was diagnosed in 51 women (28.5%), grade I microflora among 66 (36.9%) and grade II among 62 women (34.6%). In the univariate analysis, only single marital status proved to be an important risk factor associated with BV during pregnancy, this was confirmed in the multivariate analysis. Pregnant women who present risk factors for abnormal cervico-vaginal microflora should be covered by comprehensive prenatal surveillance, which enables early detection and treatment of this pathology. Research that identifies the causal pathways and mechanisms through which social disadvantage leads to a higher risk of preterm birth may help to reduce current socioeconomic and demographic disparities and improve pregnancy outcome. PMID- 12530103 TI - Evaluation of creatine kinase level during long-term tocolysis. AB - OBJECTIVE: This study was performed to evaluate serum creatine kinase (CK) levels during tocolytic therapy. METHODS: A retrospective study was performed in 27 patients who were treated with intravenous tocolytic agents for more than one week. The first-line tocolytic agent was ritodrine hydrochloride, followed by concomitant magnesium sulfate (MgSO4). The serum CK level was measured on admission and every week thereafter. The patients were divided into the normal CK (group 1) and abnormal CK (> 150 IU/L) (group 2) groups. RESULTS: Seventeen patients received both ritodrine hydrochloride and MgSO4. The CK levels in all patients rose significantly from 58.4 +/- 30.8 IU/L on admission to 116.0 +/- 68.7 IU/L on day 7 (p = 0.002). Abnormal elevation of CK occurred in 7 (25.9%) of the 27 patients. Significant differences were found between groups 1 and 2 in the total doses of ritodrine and MgSO4 (p = 0.046 and p = 0.0028, respectively). All patients in group 2 received ritodrine in combination with MgSO4 (p = 0.018). CONCLUSION: When tocolytic therapy continued for more than 1 week, nearly one fourth of patients showed an increase in CK level above the normal range. PMID- 12530104 TI - Maternal serum glycodelin in premature rupture of membranes. AB - AIM: Few studies address decidual function in abnormal pregnancies, due obviously to shortage of appropriate markers. Glycodelin is produced by epithelial cells of pregnancy decidua. We investigated whether serum glycodelin concentrations are altered in premature rupture of membranes (PROM). METHODS: Serum glycodelin concentrations were measured in 32 women with PROM and in 27 gestational age matched healthy women. Serum C-reactive protein (CRP) concentrations were measured to indicate infection. RESULTS: Glycodelin concentrations were similar in PROM patients and control subjects. No difference was found in glycodelin concentrations between the 17 PROM patients with CRP > 12 mg/l and controls, or between the 15 PROM patients with CRP < or = 12 mg/l and controls. There was no correlation between the highest antepartum glycodelin level and the interval from PROM to delivery. A negative correlation between the last glycodelin concentration and the birth weight of the newborns was found in the PROM patients (rs = -0.369, p = 0.038). CONCLUSIONS: Decidual epithelial cell function does not seem to be compromised in PROM. The inverse correlation between serum glycodelin and the birth weight of the newborns in PROM may be explained by the normally occurring decline of serum glycodelin with advancing pregnancy in the third trimester. PMID- 12530105 TI - Clinical and biophysical aspects of HELLP-syndrome. AB - OBJECTIVE: The maternal-perinatal outcome and the significance of biophysical parameters in HELPP syndrome patients were evaluated. METHODS: Sixty cases of HELLP syndrome were determined by retrospective analysis. Medical history, correlation of clinical, laboratory findings, records of fetomaternal Doppler studies, Nonstress test and maternal-perinatal outcome data were evaluated. Chi square test was used for statistical analysis, and p < 0.05 was accepted as the significance level. RESULTS: The incidence of HELLP syndrome in our institution was 1.03%. Mean gestational age at birth was 33.2 weeks, mean birth weight was 1861 +/- 710 g and mean umbilical pH was 7.25 +/- 0.13. Neonatal thrombocytopenia was demonstrable in 38% of neonates. Patients with low antepartal platelets (< 60,000/microliter) had a significantly higher incidence of intrauterine growth retarded fetuses than patients with higher platelet counts (p = 0.002). Doppler flow measurements were performed in 33 patients (55%). In 16 (48.4%) a pathological Doppler flow was documented. Doppler findings demonstrated very high sensitivity (83%) and specificity (80%) in predicting adverse outcome in growth retarded fetuses. In 17 patients (27%) fetal heart rate monitoring had an obvious pathologic pattern. Respiratory distress syndrome (74.4%) was the main indication for NICU admission. Perinatal mortality rate was 8.3% and neonatal mortality rate was 11.6%. Maternal morbidity rate was 30%. The most commonly observed maternal complications were abruptio placentae (n = 8), disseminated intravascular coagulation (n = 3) and severe postpartal bleeding (n = 3). CONCLUSIONS: In HELLP syndrome patients it is very important to closely follow maternal vital signs and fluid intake and output, and to perform fetal status assessment tests. Of the biophysical parameters, Doppler flow measurement is an especially helpful predictor of poor perinatal outcome in growth retarded fetuses in HELLP patients. Patients with very low platelets have a significantly higher risk of intrauterine growth retarded fetuses. PMID- 12530106 TI - Blood flow velocity waveforms of the fetal middle cerebral artery in a normal population: reference values from 18 weeks to 42 weeks of gestation. AB - The aim of this prospective cross-sectional study was to establish new Doppler reference curves for peak blood flow velocities (Vmax, Vmean, Vmin) and impedance indices (PI, RI) of the middle cerebral artery at 18-42 weeks of gestation by an automatic wave form analysis integrated into the ultrasound device. In 962 low risk pregnancies, blood flow velocities were derived from the middle cerebral artery with pulsed color Doppler ultrasonography. Reference curves were constructed for the individual parameters based on a growth function from a four parameter class of monotonic continuous functions according to the smallest square principle for maximum blood flow velocities, as well as on a polynomial function for resistance indices. Further assessed was intra-observer reliability. The results obtained for maximum blood flow velocities demonstrate a continuous increase (Vmax = 26.8 cm/s to 59.1 cm/s, Vmean = 11.2 cm/s to 29.7 cm/s and Vmin = 4.9 to 15.8 cm/s) over the period from 18 to 42 weeks of gestation. In contrast, reference curves for the pulsatility and the resistance index are characterized by a parabolic pattern (18 weeks: 1.5; 28 weeks: 1.9; 42 weeks: 1.1 and 18 weeks: 0.68; 28 weeks 0.8; 42 weeks: 0.61). Intraobserver reliability for PI, RI, Vmax, Vmean, and Vmax were 87%, 83%, 80%, 83% and 93%, respectively. At constant measurement conditions, the reference ranges for blood flow velocities and impedance indices in the middle cerebral artery established by this study were found to be a useful non-invasive tool in risk assessment for fetal anemia, as well as in the Doppler ultrasound observation of the course in fetuses with intrauterine growth restriction. PMID- 12530107 TI - Cerebral hemoglobin concentration and oxygen saturation measured by intensity modulated optical spectroscopy in the human fetus during labor. AB - AIMS: To quantify hemoglobin concentration and oxygen saturation in the human fetal brain using intensity modulated optical spectroscopy during labor. METHODS: A specially designed probe was applied to the fetal scalp in 20 women during uncomplicated labor at term. Optical fibres transmitted near infra-red light to and from an intensity modulated optical spectrometer (IMOS), which detected changes in the optical parameters of the infra-red light source. Using novel off line analytic techniques, these changes were converted into absolute measurements of oxyhemoglobin (HbO2) and deoxyhemoglobin (Hb). Summing Hb and HbO2 gives total hemoglobin concentration (HbT) and HbO2/HbT x 100% provided measurements of cerebral saturation (SmcO2). RESULTS: Of 20 fetuses studied, data from 10 fetuses were suitable for analysis. Over three consecutive uterine contractions, the mean (+/- SD) absolute cerebral concentrations of Hb and HbO2 were 30 +/- 18 and 46 +/ 21 mumol/l, respectively. This gave a mean cerebral HbT of 77 +/- 29 mumol/l and a mean SmcO2 of 59 +/- 12%. In the other ten fetuses insufficient light was detected to allow chromophore quantification. CONCLUSION: These are the first absolute measurements of cerebral Hb and HbO2 in human fetuses during labor. The values of total hemoglobin are similar to those obtained in neonates with hypoxia ischemia and the measurements of fetal cerebral oxygen saturation are similar to previously published values. PMID- 12530108 TI - Growth discordance and the effect of a male twin on birth weight of its female co twin: a population-based study. AB - There is an ongoing controversy whether the presence of a male twin affects the birth weight of its female co-twin, and if affected, whether birth weights of females are lower or higher in the presence of a male co-twin. The purpose of this study was to study this controversy in a large population of 12,639 twin pairs born in Israel between 1993-1998. Comparison of 7566 females in same-sex (FF) pairs with 4900 females in unlike-sex (FM) pairs, adjusted for the lighter and heavier twin in the pair, showed significantly higher birth weights of females in FM pairs. The same trend was found when concordant pairs and discordant pairs (birth weight difference > 25%) were compared separately. No significant difference was found when 7912 males in same-sex (MM) pairs were compared with 4900 males in FM pairs, adjusted for the lighter and heavier sib. Our study supports a definite intrauterine effect of the male twin on birth weight of its female co-twin. This consequence may potentially be associated with fetal programming of adult disease. PMID- 12530109 TI - Rhabdomyolysis during prolonged intravenous tocolytic therapy. AB - A patient with twin gestation was hospitalized because of preterm labor and treated with intravenous ritodrine hydrochloride and magnesium sulfate. After more than 5 weeks of therapy, the patient developed muscle pain, which was diagnosed as rhabdomyolysis. PMID- 12530110 TI - A rare presentation of Pompe disease with massive hypertrophic cardiomyopathy at birth. AB - We report a term infant with Pompe disease presenting in the immediate newborn period. The infant was born at 40 weeks' gestation, weighing 3600 g to a 32 year old black female. Infant presented at delivery with massive hypertrophic cardiomyopathy and pulmonary hypertension. Diagnosis was confirmed by low alpha glucosidase activity. The histopathology and electron microscopic findings were consistent with Pompe disease. This is the second reported case of Pompe disease presenting at delivery. PMID- 12530111 TI - Complete spontaneous resolution of severe nonimmunological hydrops fetalis with unknown etiology in the second trimester--a case report. AB - The incidence of nonimmunological fetal hydrops is between 1 in 1500 and 1 in 4000. Overall mortality for this condition is about 80%. This report describes a case of prenatally diagnosed hydrops fetalis with severe generalized edema, ascites and pleural effusion of unknown origin at 19 weeks of gestation. No symptoms were found of infections or anemia, no variabilities were observed in fetal blood analysis or in karyotyping. All signs of hydrops resolved completely after 6 weeks without treatment. The pregnancy continued with normal fetal development; labor was induced at 41 weeks of gestation. After vaginal delivery with vacuum extraction the child was completely healthy and demonstrated no signs of remaining hydrops fetalis or any other pathological parameters except an uncritical atrial septal defect type II which healed without any complications within 3 months post delivery. PMID- 12530112 TI - Epiphyseal separation of the distal humerus. AB - A case of epiphyseal separation of the distal humerus is presented, emphasizing the clinical and radiological features of this rare birth trauma. Conservative treatment resulted in complete recovery. PMID- 12530113 TI - Flow cytometric basophil activation test: a review. AB - Flow cytometry is a technique enabling the analysis of physical and biological characteristics of cells or other biological particles when labeled with antibodies coupled to fluorochromes or other dyes. The basophil activation test (BAT), also called flow-cytometric allergen stimulation test (FAST) [commercially available under the name of Flow CAST (Buhlmann Laboratories) or BASOTEST (Beckton-Dickinson)] is based on the in vitro allergen-induced specific activation of basophils. This assay rests on the demonstration of some membrane protein markers that appear after exposure to the allergen. This paper reviews some of the literature about the use of this technique in the investigation of immediate-type allergies to inhalant allergens, drugs, and foods, as well as our own experience with this diagnostic technique. Flow cytometry is a reliable method for the quantification of basophil activation after allergenic stimulus in vitro and in vivo. It also enables us to detect allergic and pseudoallergic reactions because of other mechanisms than allergen-specific IgE antibodies. Further clinical evaluation of this technique will allow validation and better establishment of its diagnostic value in allergy. PMID- 12530114 TI - Vaccines, viruses, and voodoo. AB - Vaccinations are invaluable in protection from a wide variety of diseases that can cause substantial morbidity and mortality. Although a rare complication of vaccination, autoimmune disorders represent one of these morbidities. Recently, widespread public concern has arisen from case reports suggesting that--similar to what has been observed after natural viral infections--there might be an association between specific immunizations and autoimmune diseases. Herein we address the biological plausibility of such a connection, focusing particularly on the examples of hepatitis B, rubella, and measles-mumps-rubella (MMR) vaccinations, and the autoimmune diseases they are potentially associated with. Our review of the available data suggests that, for the general population, the risk: benefit ratio is overwhelmingly in favor of vaccinations. However, the possibility cannot be ruled out that, in genetically susceptible individuals, vaccination can result in the unmasking of an autoimmune disease triggered by the immunization. We also critically examine the existing data suggesting a link between immunization against MMR and autism, and briefly discuss the controversial evidence pointing to a possible relationship between mercury exposure from vaccines and autistic disorders. There is a continued urgent need for rigorously designed and executed studies addressing these potential associations, although the use of vaccinations remains a critical public health tool for protection against infectious disease. PMID- 12530115 TI - New insights on latex allergy diagnosis and treatment. AB - Because of widespread latex manufacturing in the last decades, exposure to latex has become ever greater, as has the incidence of latex allergy among the general population as well as the high-risk groups. Clinical manifestations range from local reactions (edema, itching, erythema, and papulae), rhino-conjunctivitis, asthma, pharyngeal edema, to severe systemic reactions such as anaphylactic shock (from parenteral exposure especially during medical examinations or surgical procedures). Diagnosis is formulated on the basis of the personal history and an accurate allergological evaluation. Patients usually have a history of food intolerance or adverse reactions during medical or surgical procedures, which are believed to be the result of anesthetics or other drugs. In vivo tests are prick and patch tests and challenge tests. Challenge tests (nasal, conjunctival, bronchial, intravaginal, sublingual, oral, and cutaneous tests) are important for confirming the diagnosis, for evaluating a patient's response to the different kinds of latex exposure, and for verifying the effects of desensitizing treatments on the various organs involved in latex allergy. The most important in vitro test is the specific anti-natural rubber latex IgE assay. The only effective resolution to the latex allergy problem seems to be desensitization. The criteria of effectiveness are the capability to use latex items, to undergo medical examinations (even invasive), and to remain in an environment where latex particles can be inhaled (such as surgical departments), without symptoms. The first attempts were performed by subcutaneous administration of latex, but this protocol seems to be delicate to perform because of the side effects. Two different alternative methods (percutaneous and sublingual) were successfully performed and proved to be safer and more effective, even though further studies on a larger group of patients are needed. PMID- 12530116 TI - In vivo and in vitro sensitization to domestic mites in German urban and rural allergic patients. AB - Sensitization to domestic mites is common in Germany. The main objectives of this study were (1) to establish the rate of skin test sensitivity to Dermatophagoides pteronyssinus, Acarus siro, Lepidoglyphus destructor, and Tyrophagus putrescentiae in 512 consecutive patients evaluated for upper and/or lower respiratory complaints; (2) to verify how many of the patients with a positive skin test to at least one of the three storage mites were also skin test positive to D. pteronyssinus; and (3) to verify how many of the patients with at least one positive skin test to one of the storage mites previously mentioned were also sensitized, in vitro, to other mite species. A total of 512 consecutive patients with rhinitis and/or asthma, living in urban or rural areas of central Germany were skin tested with extracts of D. pteronyssinus, A. siro, L. destructor, and T. putrescentiae. In addition, specific IgE determinations to Euroglyphus maynei, Blomia tropicalis, Blomia tjibodas, Blomia kulagini, and Gohieria fusca were conducted in those individuals with a positive skin test to at least one of the storage mites used in skin testing. Of the 512 patients, 103 (20.1%; 77 urban dwellers and 26 farmers) reacted to at least one of the storage mites. From this latter group, 88 individuals (85.4%) also skin tested positive to D. pteronyssinus. In vitro specific IgE determinations revealed a high rate of sensitization to the other mite species studied. We conclude that sensitization to storage mites in Germany is frequently associated with sensitivity to D. pteronyssinus. Overall, skin test sensitivity to storage mites was greater in rural than in city dwellers. In vitro sensitization to B. tjibodas was also significantly greater in rural than in city dwellers. PMID- 12530117 TI - Relevance of the selected cytokine release (TNF-alpha, IL-6, IFN-gamma, and IFN alpha) to the exacerbation of bronchial asthma from airway mycotic infections. Predominant role of TFN-alpha? AB - Airway fungal infections are often associated in asthmatics with the exacerbation of asthma symptoms. However, the pathomechanism of this phenomenon has not been fully understood. The aim of our study was to assess whether antimycotic treatment can influence the capacity of bronchoalveolar (BAL) leukocytes to release proinflammatory cytokines, which could contribute to increase in asthma severity. Ten patients with bronchial asthma complicated by airway fungal infections (Candida albicans and/or Aspergillus fumigatus) were included in the study. Seven asthmatics were treated with systemic and inhaled corticosteroids, whereas the remaining three with inhaled ones only. All subjects underwent several courses of therapy with antibiotics due to respiratory infections. BAL leukocytes obtained from the patients were cultured in the absence or presence of lipopolysaccharide E.coli (LPS) or Newcastle disease virus (NDV). The BAL procedure and measurement of the levels of tumor necrosis factor-alpha (TNF alpha), interleukin-6 (II-6), interferon-gamma (IFN-gamma), and interferon-alpha (IFN-alpha) by specific bioassays were performed twice: before antimycotic treatment and after 3 weeks of therapy with 8 mg of nebulized fluoconazole and 400 mg of oral ketoconazole per day. The elimination of fungi from respiratory tract resulted in an apparent clinical improvement. This coincided with diminished production of TNF-alpha in response to LPS and the production of IFN alpha in response to NDV, which were initially high and subsided significantly after antimycotic therapy (p = 0.035, and 0.011, respectively). Such changes were not observed in the case of IFN-gamma and IL-6. This may suggest that TNF-alpha as well as IFN-alpha are secreted by fungi-prestimulated leukocytes from the lower respiratory tract and may be involved in the processes of exacerbation of asthma complicated by fungal infections. Further analyses of relationships between changes in cytokine levels and clinical parameters indicated that IFN alpha seems to be of particular interest in fungal stimulation of asthma. PMID- 12530118 TI - Lack of association between atopic asthma and the tumor necrosis factor alpha-308 gene polymorphism in a Czech population. AB - Susceptibility to the development of asthma and other atopic diseases is known to be associated with genetic components. Several investigators have linked the tumor necrosis factor (TNF) genes and nearby markers located on chromosome 6p to atopy and asthma. A recent study has demonstrated that the TNF-alpha*2 allele of a polymorphism in the TNF-alpha gene promoter region (G-308 A) is associated with a higher risk for the development of atopy in Spanish patients. This study evaluates the possible role of two described bi-allelic polymorphisms in the TNF locus [a G to A transition at position-308 in the 5'-promoter region of the TNF alpha gene and an NcoI restriction fragment length polymorphism (RFLP) in the first intron (+252A/G) of the LT-alpha(TNF-beta) gene] in atopic diseases in a Czech population. We investigated the distribution of these polymorphisms in a case-control study. The genotypes were determined in 151 patients with atopic asthma and 155 randomly sampled control subjects. The genotype frequencies for both polymorphisms were similar in cases and controls. No significant differences in allele frequencies were found between either of the patients groups and the reference subjects. Similarly, there were no associations of any of the examined variants of the TNF genes with total IgE, specific IgE or pulmonary function tests in patients with allergic diseases. We conclude that these polymorphisms of the TNF genes are unlikely to contribute to atopic disease risk in our population. Significant associations that have been reported in other studies may reflect the genetic heterogeneity of these complex diseases. PMID- 12530119 TI - Do the molecules CD26 and lymphocytes activation gene-3 differentiate between type 1 and 2 T cell response? AB - BACKGROUND: The Th1/Th2 paradigm is considered to be responsible for the development of many immunological disorders, including atopic diseases and diabetes mellitus type 1. So far, however, no unequivocal markers identifying the two subpopulations of T cells have been found. OBJECTIVE: To examine the expression of two putative markers of Th1-mediated disorders, CD26 and lymphocytes activation gene-3 molecules, on CD4+ and CD8+ lymphocytes in the peripheral blood of atopic and diabetic donors. METHODS: 11 patients (9 males/2 females, age range: 21-29, median age: 23) suffering from episodic atopic bronchial asthma (AO group) and 11 patients (9 males/2 females, age range: 33-47, median age: 42) with insulin-dependent diabetes mellitus (DM group) were included into the study. Atopy was excluded in all diabetic patients. The following T cell subsets were analyzed by flow cytometry (FAC-Scan): CD8+CD26+, CD4+CD26+, CD4+LAG 3+, CD8+LAG-3+, and CD56+. Nonparametric tests were used for statistical analysis, and results were expressed as median and quartile range. RESULTS: Among all analyzed T cell subsets, the number of CD4+LAG-3+ and CD8+LAG-3+ cells was significantly higher in the A0 group--8.3% (5.4-11.7) vs DM 5.4% (3.9-5.9), p < 0.05, and 13.3% (8.8-19) vs DM: 7.1% (6.2-8.3), p < 0.008, respectively. The ratio between CD4+/CD8+ subsets was similar in both groups: CD4+CD26+/CD8+CD26+- AO: 7.2 (5.3-10.1); DM: 6.4 (5.5-12.6), p = 0.77 and CD4+LAG-3+/CD8+LAG-3+--AO: 0.64 (0.58-0.67); DM: 0.71 (0.64-0.78), p = 0.082. CONCLUSION: The results suggest that the two examined putative markers of type 1 T cells do not discriminate between Th1- and Th2-dominated responses. PMID- 12530120 TI - Psychological status of patients with seasonal and perennial allergic rhinitis. AB - There has been a tendency among some clinicians to underestimate the psychological problems of patients with rhinitis. This study assesses the psychological status of allergic rhinitis patients and the influence of type, duration, and severity of rhinitis on the psychiatric evaluation. Forty-one patients with seasonal and perennial allergic rhinitis and 36 healthy control subjects were enrolled in the study. All patients and control subjects were studied through psychological interviews and several psychological tests such as Symptom Checklist-90 (SCL-90), Courtauld Emotional Control Scale, the McMaster Family Assessment Device, and the Satisfaction with Life Scale. Statistically significant differences between patients and controls were observed in all subscales of the SCL-90, particularly in somatization, depression, and general symptom index subscales. Differences between groups were not significant for the other psychological tests. Additionally, differences between seasonal and perennial allergic rhinitis patients were not significant in all questionnaires. Total symptom scores of rhinitis were positively correlated with general symptom index, somatization, interpersonal relationship difficulties subgroups of the SCL 90, whereas negatively correlated with the scores of satisfaction with the life scale. Patients with allergic rhinitis--regardless of type of rhinitis--may develop a group of psychological complaints related to symptom severity. Since the recognition of specific psychiatric disorders can have therapeutic importance, further studies focusing on the associations of these disorders and allergic rhinitis should be conducted. PMID- 12530121 TI - Elevation of serum eosinophil cationic protein, soluble tumor necrosis factor receptors and soluble intercellular adhesion molecule-1 levels in acute bronchial asthma. AB - Asthma is a disease characterized by chronic airway inflammation. Many inflammatory cells and molecules contribute to its pathogenesis. Soluble tumor necrosis factor receptors (sTNFRs) and soluble intercellular adhesion molecule-1 (sICAM-1) play regulatory roles in the inflammation. But their roles in the inflammation of asthma have not been well defined. This study was done to examine the changes in serum levels of these molecules in acute asthmatic patients. The concentrations of eosinophilic cationic protein (ECP), sTNF-RI, sTNF-RII, and sICAM-1 were measured in sera of 24 asthmatic patients during acute attack, in 18 patients at 24 h, and in 10 patients at 7 days after attack and in sera of 14 healthy control subjects by ELISA method. Serum levels of ECP, sTNF-RI, sTNF-RII and sICAM-1 in the patients with asthma during attack were significantly higher than those of the controls (p < 0.001, p < 0.01, p < 0.05, p < 0.05 respectively) and stayed high up to the 7th day. In conclusion, high serum levels of sTNF-RI, sTNF-RII and sICAM-1 suggest that these molecules may contribute to the regulation of allergic inflammation and may reflect the severity of inflammation in the airway of asthmatic patients. PMID- 12530122 TI - Ultraviolet recall phenomenon following the use of ampicillin. AB - Ultraviolet recall or sunburn reactivation is an uncommon phenomenon that has been most frequently reported in patients who suffered sunburns and then subsequently underwent therapy with methotrexate. There are very few cases in which antibiotics are involved. We report a case of ultraviolet recall occurring after ampicillin therapy. Our patient suffered severe vesiculobullous skin reaction in areas that had been sunburned only minimally 2 months previously. The patch test was positive only to ampicillin tested in the skin previously affected. PMID- 12530123 TI - Intrinsic factors for unbound theophylline. AB - The developmental changes of the ratio of unbound theophylline to total serum concentration have been demonstrated, suggesting clinical usefulness. However, the molecules that can replace the theophylline binding sites of albumin could influence theophylline-binding affinity. We evaluated the intrinsic factors influencing the unbound theophylline ratio. The results suggested that physicians should note that a more intense response might be obtained in children with various intrinsic factors even at the same total theophylline concentration. PMID- 12530124 TI - Transplant trauma. New tactics fight the growing organ donor deficit. PMID- 12530125 TI - The high cost of eggs. PMID- 12530126 TI - The sound of unsound minds. Great music is linked to troubled composers, a psychiatrist-pianist says. PMID- 12530127 TI - Clowns or cloners? The strange genesis of a human cloning claim. PMID- 12530128 TI - Booty camp. The fitness biz has bold new ways to trim your butt (and build muscles). PMID- 12530129 TI - [International conference "Genetic Consequences of Emergency Radiation Situations" (Russian Federation, Moscow, 10-13 June 2002]. PMID- 12530130 TI - Relevance of radiation-induced bystander effects for environmental risk assessment. AB - A novel mechanism involving a medium borne signalling factor has been identified following irradiation of populations of cells to doses ranging from 5 mGy-5 Gy gamma-rays or to as little as 1 alpha particle traversal in a culture containing hundreds of cells. The factor can be released into culture medium and can induce responses in unexposed cultures. It has been called a "radiation-induced bystander factor". The effect is obviously relevant to risk assessment as it happens at very low doses. It could also offer new avenues for development of drugs aimed not at cell destruction but at restoring the tissues own control and coordination of response following DNA damage. The effect is clearly induced by radiation and probably by other substances. While these effects are now accepted to happen both in vitro and in vivo, their relevance and function is unknown. The investigation and modelling of the mechanism and the variation in level and type of effect in relation to genetic background and clinical history are key questions which need to be addressed in the field. The key driving hypothesis of the work being done by our laboratory is that radiation-induced bystander effects (RIBE) reflect emergent control in complex tissues and communicating cell systems, which can be harnessed for therapeutic gain. PMID- 12530131 TI - [Molecular genetics of radiation-induced chromosome breaks in a gene area in Drosophila: "position" effect of gene mutation?]. AB - On the sample of 43 gamma-ray and neutron-induced inversion or translocation exchanges with the vestigial (vg) phenotype, the molecular cytogenetic analysis of distribution of exchange breakpoints on the molecular map of Drosophila vg region (subsection 49D3-4 on the polytene chromosome 2R) was performed using hybridisation in situ technique. Simultaneously, PCR-assay of DNA alterations in all exons and introns (except for intron 4) of the vg gene for 18 mutants with exchange breakpoints outside of the gene was carried out. The results obtained by these molecular genetic techniques have shown that 1) radiation-induced breaks under chromosome exchanges with the vg phenotype were regularly located inside of the vg gene (19 cases out of 43 studied ones or 44.2%) passing through the large introns; 2) breakpoints were frequently flanked by deletions of the gene as whole (3 exchanges) or of its major part (3 exchanges); 3) many of the breaks (18/43 or 41.8%) are situated outside (distal or proximal) of the gene although such mutants have got the vg phenotype; 4) 2/3 (12/18 or 66.7%) vg mutants with the breakpoint outside of gene show the intragenic DNA lesions (microdeletions, microinversions) occurring obviously independently and simultaneously with the neighbor chromosome breaks; 5) only each third vg mutant with break outside of the gene (6/18 or 33.3%) have the unchanged gene subregions under study and presents obviously the result of "position effect" which appear to manifest itself for a distance of 2-30 kb (more near and farther locations of the proximal and distal breakpoints, respectively, relative to the vg gene). Our findings showing regular induction of the multiple genetic lesions (chromosome breaks and mutations of the adjacent genes) on the both ends of chromosome exchange induced by single track produced by gamma-rays or neutrons were discussed as a scientific basis for the conceptually new approaches to the assessment of both genetic damage numbers in the cell genome with chromosome exchange (the multiple genetic lesions) and radiation genetic risk (our molecular genetic approach showing the need for an increase of risk levels at least on a factor of 3 for the heritable chromosome alterations detected by the ordinary cytogenetic monitoring). PMID- 12530132 TI - Radiation sensitivity and the status of some radiation sensitivity markers in relatively sensitive lymphoid cells. AB - The most commonly used indicators of ionizing radiation exposure are cytogenetic measures and survival parameters. All these methods have their advantages, disadvantages and uncertainties, such that better biological estimators of the absorbed dose, especially in the low dose range, are being sought. In this study we analyzed apoptosis and several proteins involved in the regulation of apoptosis as possible indicators of irradiation after relatively small doses (0.1 2 Gy) of X-rays. The studies were carried out in seven lymphoid cell lines: two mouse lymphoma L5178Y, the human pre-B cell leukemia Reh, and four human Epstein Barr virus-transformed lymphoid cell lines (two apparently normal and two Ataxia telangiectasia (AT)). We detected apoptosis with the in situ terminal deoxynucleotidyl transferase assay and flow cytometry, and measured the expression of several apoptotic-regulatory proteins (Bcl-2, Bax, Bclx, NF kappa B) with Western blotting. The cytokinesis-block micronucleus assay, comet assay as a measure of DNA damage, and trypan blue survival test were also done for comparison Although for the most of examined parameters of radiation sensitivity: i.e. micronucleus assay, trypan blue test and percentage of apoptosis--there were observed clear dose-effect relationships for all cell lines examined, we did not find agreement between values for these measured parameters. There are marked differences in both timing of apoptosis and percentage of apoptotic cells. Variation in the apoptotic fraction in the controls for different sets of experiments is not very pronounced. There is however considerable variation for the same parameters in irradiated cells, possibly due to their cell cycle status during irradiation, as the cultures were not synchronized. Overall, neither the numbers of apoptotic cells nor the expression of apoptosis-related proteins, nor DNA repair can serve as dose estimators or sensors for these lines, but still these parameters can give valuable supplementary information about radiation sensitivity. PMID- 12530133 TI - Acceleration of DNA-double strand rejoining during the adaptive response of Chlamydomonas reinhardtii. AB - Newly developed constant-field low voltage electrophoresis (adapted for algae cells by us) was applied to quantify the induction and repair of nuclear DNA double-strand breaks, by measuring the movement of DNA out of the starting wells into the electrophoresis gel using a UV-gel scan and computer analysis of DNA ethidium bromide fluorescense (Syngene; Gene tools). A cell-wall-less mutant strain of Chlamydomonas reinhardtii (CW15) was used; the DNA and proteins are easily accessible because of the lack of an outer cell wall. Our results showed that giving a small priming dose (50 Gy) led to a small acceleration of dsb rejoining. When the magnitude of the priming dose was progressively increased, there was a corresponding decrease in the fraction of damage remaining at 4 hours after radiation exposure (to a test dose of 500 Gy). This indicates an upregulated rejoining of dsb following exposure of cells to the priming dose, which may be related to the strong adaptive response in this organism. Protein synthesis inhibitors were found to reduce the rate of rejoining of dsb, and from earlier results are known to inhibit the adaptive response. Thus, the adaptive response is likely to be dependent on increased dsb rejoining and depends on de novo protein synthesis. The nature of these proteins has not yet been established. C. reinhardtii CW15 is an attractive model system in which to study the underlying mechanisms of the adaptive response to ionizing radiation, and its underlying link with dsb rejoining. The results are interesting both from a basic biological point of view, and as a means to further understand the response of tumour cells to radiation therapy since the adaptive response has been postulated to determine the shape of the "shoulder" region of the survival curve of cells at low doses of radiation. PMID- 12530134 TI - Interphase chromosome locus displacement induced by low-doses of radiation. AB - The most important stage in the making of mutations is a reparation of different DNA damage, including the more deleterious double-strand DNA breaks (DSB). The first stage of adaptive response--fundamental antimutagenic cell reaction, purposeful to reparation for induced DSB repair--is investigated in present work. Non-radioactive in situ hybridization of biotin-labeled DNA probe was used to mark chromosome 1 pericentromeric regions (PR) in G0 human lymphocytes. It was shown that under 3-10 cGy (X-radiation, 160 kV) PR become displaced from a nucleus periphery to inner territory of a nucleus. The moving process realizes during several hours after an irradiation. As far as some non-specific gene repressors are co-localized with chromosome centromeric regions it is possible hypothesizes that the displacement cause changing expression of some genes. It is possible to propose that an absence of radiation induced chromosome locus displacement may be one of causes DSB repair disturbance. This hypothesis was tested by the model. It is assumed that one consequence of the underlying defect may be inappropriate involvement of cell's recombination machinery in the repair of DSB. We studied lymphocytes of patients with hereditary BRCA2 mutation. It is thought that this gene takes part in DSB repair. The significant differences of the PR moving between control samples and the cases were revealed under 10 cGy. Similar results were observed on lymphocytes of patients with Fanconi syndrome. Thus, abnormal moving of interphase nucleus chromosomes conditioned by low-dose irradiation may suggest on imperfect machinery of DSB repair, i.e. genetic risk. We realize that further investigations are needed for definitive conclusion. PMID- 12530135 TI - [Detection of genomic instability in offspring of male mice chronically exposed to gamma-radiation by the "adaptive response" test]. AB - The genomic instability (GI) in somatic cells of the progeny (F1 generation) of male mice chronically exposed to low-dose gamma-radiation was studied by comparative analysis of chromosome damage. BALB/C male mice exposed to 0.1 Gy (0.01 Gy/day) and 0.5 Gy (0.01 and 0.05 Gy/day) were mated with unirradiated females 15 days after irradiation. For comparison of radiosensitivity, two-month old males, the descendants of irradiated and unirradiated animals, were subjected to irradiation with a dose of 1.5 Gy (0.47 Gy/min) from a 60Co source. GI was revealed by the standard scheme of adaptive response. The experiments indicated that, by using the test "adaptive response", it is possible to detect the transition of gamma-radiation-induced genomic instability in sex cells of male parent into somatic cells of mice (F1 generation) either from changes in radiosensitivity or by the absence of the adaptive response induced by a standard scheme. PMID- 12530136 TI - [Mathematical analysis of genetic effects of low doses of ionizing radiation]. AB - The comparative study of effects of low doses of radiation on peripheral blood lymphocytes of persons occupationally exposed to radiation and non-exposed ones was carried out. The main attention was paid to radio-adaptive response forming under consistent exposure to low (0.05 Gy) and damaging (2 Gy) doses of gamma irradiation. Noticeable heterogeneity in capacity for adaptive response forming in occupational group was revealed. The mathematical model adequate to experimental material was constructed using Kohonen neuronets. PMID- 12530137 TI - [Biochemical mechanisms of radiogenic cytogenetic and somatic disorders in children residing in regions polluted by radionuclides]. AB - Children and adults affected by the Chernobyl accident suffer with lipoperoxic stress coupled with hypovitaminoses A and E, the syndrome and its radiogenic mechanisms described by the authors previously. The relation of these biochemical disturbances to somatic consequences on mutagenic and teratogenic levels caused by chronic low-dose irradiation of children into radiopolluted regions has been investigated. Either the rational approaches to prevent or/and to treat all pathologies indicated were tested. The radionuclide polluted regions had 1-5, 5 15 and 15-40 Ci/km2 by 137Cs. Unprotected newborns (without the therapy) had vA and vE levels about one third of normal ones for minimal radioloads, the vitamins lowered further for higher Da and/or Dm up to the deep hypovitaminoses or even avitaminoses. Increasing of LPC catabolites levels for children were correlated significantly with their stigmation levels. The therapy-protected newborns had normal ranges of all indices. However, unprotected newborns showed dramatic increase for all of radiogenic indices along with fall of vA and vE, and significant inverse correlations of the indices with retinol levels. The values of radiogenic indices showed hyperbolic-like rise along with growing of vA and vE deficits below their lower limits. The combination of radiogenic biochemical deviations evaluated for Chernobyl contingents is supposed to be the primary molecular mechanisms of somatic mutations and irreversible stigmation. The peroral polyvitamin therapy is proposed as the method of choice for prevention or normalization of revealed pathologies. PMID- 12530138 TI - [Mutation at the T-cell receptor locus in people a long time after acute and prolonged irradiation]. AB - The frequency of lymphocytes with mutations at genes of T-cell receptor (TCR) is determined in 165 persons exposed to ionizing radiation 16-40 years after the influence. Depending on a type of irradiation and time, elapsed from the moment of exposure, all inspected persons were divided into 3 groups: 1) the analysis performed 16-40 years after acute irradiation; 2) the analysis--9-13 years after acute irradiation; 3) the analysis--9-13 years after prolonged irradiation. The persons with elevated frequencies of TCR-mutant cells were detected in all three groups. In the first group they have compounded 36% of persons inspected, in the second--25%, in the third--15%. The percentage of persons with elevated frequencies of mutant cells in all groups was significantly higher than in control groups. Taking into account fast enough elimination of radiation-induced TCR-mutant cells it is possible to suspect, that the cause of elevated frequency of mutant cells in the part of the persons can be genome instability. PMID- 12530139 TI - [Coding in molecular biology and radiation]. AB - In work, on the basis of known base biological laws, the new natural three dimensional designs are under construction: codonogram and aminogram. Codonogram and aminogram have revealed the new laws of coding and constructions in molecular biology. The secondary coding codonogram, at the expense of redundancy, has enabled for DNA, RNA, and mRNA to construct the hidden coding layers (HLC). HLC are constructed on a base of pirimidine and purine. In interaction with ferments HLC, should guarantee: addressation, synchronization, correction and other functional transformations of a gene. Radiation can damage a layer mRNA of synthesis protein, hidden layers of coding and structure of the ferments of the cell. In work are given: kodonogram, aminogram UGC, basis of construction HLC, the references to the literature with an example of decoding. PMID- 12530140 TI - [Ability of energy metabolism of cells to activate repair of its genetic material after radiation damage]. AB - A new hypothesis on a mechanism of action of the radioprotective and medical preparations involving the activation of oxidative phosphorylation and biogenesis of mitochondria which provides the increase of dimension and rapidity of the cellular reparation has been suggested. A previously unknown property of mitochondrions to determine a cellular homeostasis has been found. An activation of oxidative phosphorylation of mitochondria is a way of cellular protection against extreme conditions (namely, against radiation exposure and tumor formation). PMID- 12530141 TI - [Regularities of excising transposon Tn10 in rec-mutant E. coli cells exposed to gamma radiation]. AB - The regularities of gamma-induced excision of transposon Tn10 in different rec strains of E. coli cells after gamma-irradiation have been studied. The survival of cells and relative frequency of the Tn10 elimination as a function of the 137Cs gamma-radiation doses were investigated. RecN and recA-mutants of E. coli were used for study of the role of rec-genes in the gamma-induced transposon excision. It was shown that the induced excision in the recN mutant was reduced. The transposon excision in the recA mutant was not revealed. The obtained results let to conclude that recA, and recN genes are involved not only in DNA repair processes but also in the gamma-induced transposon excision in bacteria. PMID- 12530142 TI - [Genetic and somatic effects of ionizing radiation in humans and animals (comparative aspect)]. AB - Genetic effects of ionizing radiation in the progeny of exposed parents could be conventionally subdivided on three main types. 1. Severe developmental disorders (fetus death, stillbirth, early postnatal mortality, malformation, hereditary disease, sterility). These effects are known to be caused by so called "gross" mutations (genomic, chromosomal, those of essential genes) with dominant harmful effects. Although found in rodents, insects, fishes they have not been found in humans. The proposal is made that the higher reproductive potency of a species the higher its tolerance for defective organisms. Due to strong selection against severe defects at the earliest stage of pregnancy, this genetic effect of radiation is likely to be difficult to detect in people. 2. Increased cancer risk manifested as elevated incidence of spontaneous tumors and increased sensitivity to carcinogenic agents. 3. Decreased fitness (non-carcinogenic negative health effects). These two last types of radiation genetic effects are presumably due to instability and functional inferiority of cell genome in the progeny of irradiated parents. The genetic background of these effects is suggested to be the load of induced minor mutations in regulatory genes (mini-, microsatelite loci) or/and epigenomic rearrangements of DNA in parental germ cells transmitted to progeny. These radiation genetic effects are much more obvious in animals as compared to humans. Apparently, they are difficult to find in humans because of their essential dependence on promotive (life style) factors, which are impossible to control in the offspring of irradiated people. A comparison of somatic (in irradiated organisms) and genetic (in the progeny of irradiated parents) effects of radiation provide evidence on the phenomenological as well as pathogenic similarity. PMID- 12530143 TI - Radioprotective effect of novel disubstituted thioureas on pea (Pisum sativum L.) development. AB - The review presents our research on the influence of novel thiourea compounds on the biological and genetic effect of gamma-rays using in vivo and in vitro systems of pea. Some novel disubstituted thioureas: o-allylthioureidobenzoic acid (ATB); o-phenylthioureidobenzoic acid (PTB); N-allyl-N'-2-pyridylthiourea (A-2 PTU); N-phenyl-N'-2-pyridylthiourea (P-2-PTU) and 1,4-allylthioureidosalicylic acid (ATUS) were examined. Pea (Pisum sativum L.) seeds from five varieties were used. Experiments in vivo and in vitro were carried under laboratory, greenhouse and field conditions. The data revealed the PTB radioprotective effect demonstrated by: reduction of chromosome aberrations by 2 folds; 50% increase of germinating and surviving plants in M1; twice higher frequency of induced mutations in M2 generation relative to irradiation without PTB treatment; decreasing the level of induced radiation suppression leading to favorable effect on the initial stem and root development of pea. ATB radioprotective effect was demonstrated in vitro by: 25-35% stimulation of organogenesis; by 20-50% increase in bud formation; by 25% stimulation of growth. The effect of A-2-PTU and P-2-PTU depended on the irradiation dose. The protective effect of A-2-PTU is more pronounced at lower irradiation dose, while the effect of P-2-PTU is more pronounced at higher irradiation dose. ATUS, opposite to the other compounds, revealed radiosensibilizing effect by: 16-27% increase in lethality caused by gamma-rays leading to lower number of germinating and surviving plants in M1; 50% decrease in the number of induced mutations in M2 generation; limiting the types of induced mutations at the higher irradiation dose. As a result of the experiments useful mutation forms were obtained, characterized with: earliness, lodging and disease resistance; higher productivity. PMID- 12530144 TI - [Genetic efficacy of low doses of ionizing radiation in chronically-irradiated small mammals]. AB - Earlier we have established the genetic effects of low dose chronic irradiation in bank vole (somatic and germ cells, embryos), in pond carp (fertilized eggs, embryos, fry) and in laboratory mice (somatic and germ cells) in the range of doses from near-background to 10 cGy. These low dose effects observed in mammals and fish are not expected from extrapolation of high dose experiments. For understanding reasons this discrepancy the comparative analysis of genetic efficiency of low dose chronic irradiation and the higher doses of acute irradiation was carried out with natural populations of bank vole which inhabited the two sites differing in ground of radionuclide deposition. For comparing efficiency the linear regression model of dose-effect curve was used. Dose-effect equations were obtained for animals from two chronically irradiated bank vole populations. The mean population absorbed doses were in the range 0.04-0.68 cGy, the main part of absorbed doses consisted of external radiation of animals exposed to 137Cs gamma-rays. Dose-effect equations for acute irradiation to 137Cs gamma-rays (10-100 cGy) were determined for the same populations. Comparison of genetic efficiency was made by extrapolation, using regression coefficient beta and doubling dose estimation. For chronic exposure the doubling doses calculated from low-dose experiments are 0.1-2 cGy and the doubling doses determined from high-dose experiments are in the range of 5-20 cGy. Our hypothesis that the doubling dose estimate is calculated in higher-dose ionizing radiation experiments should be much higher than the deduced from the low dose line regression equation was verified. The doubling dose estimates for somatic cells of bank vole and those for germ cells of laboratory mice are in close agreement. The radiosensitivity of bank vole chromosomes were shown is practically the same as that for human lymphocytes since doubling dose estimates for acute irradiation close to each other. For low LET radiation a higher genetic efficiency of chronic low doses in comparison with the higher doses of acute gamma-irradiation (137Cs source) was proved by three methods. PMID- 12530145 TI - [Modern problems of biodosimetry]. AB - Humans have different radiosensitivity. Besides, many factors which are not under control can change significantly biological effects of radiation. Some of substances are present in our food--residual amounts of fertilizers or herbicides can be mutagenic or influence mutagenic action of radiation. In the last case synergetic or antagonistic effects can be observed. At the same time many food products contain radioprotectors or antimutagens. Some drugs, stress, virus diseases and so on can change biological effects of radiation too. Radioadaptive response is one of the most significant factors which can be responsible for incorrect radiation dose evaluation. All these uncontrolled factors may be responsible for significant mistakes of evaluating radiation dose by biological methods. That is why biological methods can be used for bioindication but not for biodosimetry. PMID- 12530146 TI - [Chromosomal instability in rodents from the EURT territory: interspecies comparison]. AB - The common vole (Microtus arvalis) and the mole vole (Ellobius talpinus) were trapped in the Eastern Urals Reserve (initial soil pollution 500-1000 Ci/km2) where many dozens of generations came to pass after the 1957 accident in the plant "Mayak" and in the Middle and South Urals and Transurals localities with global level of the 90Sr pollution. At present the average internal dose rate from 90Sr was 0.413 mGy/day in M. arvalis and 0.985 mGy/day in E. talpinus. The total accumulated doses from external and internal sources were 0.4-0.5 Gy per year. Frequencies of cell with chromosome aberrations and gaps and rates of aneuploidy and polyploidy were analyzed. In the common vole from EURT, the frequency of aberrant cells was significantly higher (4.57%) than in three control populations (0.44-0.86%). In the mole vole, all the cytogenetic parameters did not exceed the control ones. Absence of the genomic response in E. talpinus could be caused by the protective activity of melanin-related metabolites or by heritable factors, which may have arisen as a result of mutagenic pressure and have spread in the population in question both by selection and genetic drift, due to considerable isolation of local demes of E. talpinus. PMID- 12530147 TI - [Dynamic component of maintaining genomic stability in murine bone marrow cells after chronic low-intensity irradiation lasting one year]. AB - For analysis of a dynamic component state of the system of maintenance of genome stability, which represent a condition of its expression (first of all, genes of the control of phases of cell cycle, the DNA repair and redox systems) after a long chronic exposure to a small dose, the activity of replicative, reparative DNA synthesis, DNA damage as well as oxyradical content in the bone marrow cells of mice (critical for radiation effects mammalian system) after 1 year radiation exposure to a dose 63.7 cGy (0.17 cGy/day) were studied. The considerable enhancement of replicative and reparative DNA synthesis activity by 67% (p = 0.0033) and 60% (p = 0.000004) accordingly in relation to the control and some, but statistically not significant (p = 0.149) tendency to increase (by 30%) the content of a superoxide anion-radical were established. Strong and highly significant correlation (r = 0.8681; P = 0.99975) between DNA damage and O2-. content in bone marrow cells of the irradiated mices, which indicate the large DNA damage by oxiradicals, probably, due to the loss of a part of structural proteins and conformation changes in expression sites of a chromatin, were detected. The obtained results interpreted as representing the change of a dynamic component state of a system of maintenance of genome stability, the epigenic transfer of that to descendants of the irradiated cells can be the cause for formation and maintenance of radiation-induced genome instability. PMID- 12530148 TI - [Cytogenetic effects of 241Am in the Allium test]. AB - The purpose of this study was to elucidate the genome damage induced by 241Am irradiation using different parameters of cytogenetic evaluation in Allium-test. The root tip cells test-system for the cytogenetic effects studying was used. 241AmCl3 of different concentrations was used (1.5 x 10(-9)-1.5 x 10(-7) g/l). Water solution-to-plant transfer factor for 241Am was found to be 0.18 +/- 0.04. The internal doses of 241Am accumulated during germination were 0.37-37.00 cGy. The impact of 241Am-irradiation was evaluated on the mitotic index (MI), the yield of aberrant anaphases (AA), the distribution of chromosome aberrations number in cells and the average level of lesion of aberrant cell (LAC). Probably all these parameters are differ in sensitivity to damage factor, but only some changes in MI was revealed. It is supposed, that the absence of any changes in the distribution of chromosome aberrations number in cells and the average level of LAC in 241Am-irradiated cells confirm the absence of significant 241Am-impact on chromosomes, as the alpha-irradiation should cause significant damages in chromosomes. Although solutions of 241Am were high-concentrated, the seedlings didn't accumulate high internal doses. It appears the distribution of 241Am is a significantly heterogeneous hence it is possible the absorbed doses in nuclei can't reach the level necessary for revealing of cytogenetic effects. PMID- 12530149 TI - [Down syndrome and ionizing radiation: causal effect or coincidence]. AB - Annual and monthly prevalence of Down's Syndrome has been analyzed in Belarus for a 19-year period (1981 to 1999). The distribution of 2786 DS cases during this period showed no significant long-term prevalence trend. Meanwhile, 2 marked increases of DS prevalence were registered in 1987 and 1990. The most remarkable annual increases were observed in 1987 in Minsk city, in Vitebsk oblast (considered as contamination-free administrative district) and in Minsk oblast (partially contaminated area), data not shown in the text. Much more marked and significant increases were recorded in January 1987, in Minsk city, Gomel oblast and Minsk oblast. However it is impossible to establish a causal relationship between these clusters of DS and exposure to the Chernobyl fallout, at a geographical level. The presence of such a cluster 9 months after the explosion in one of the most exposed area (Gomel oblast) should incite to careful conclusions and prompt further studies. PMID- 12530150 TI - [Some complexities in evaluating radiation-induced mutagenic effects in man]. AB - Chromosomal aberrations in somatic cells (circulating lymphocytes) have been investigated in a remote period in different groups of people who suffered from radiation accidents. The chromosomal radiation markers were found even decades after irradiation. The question about the necessity of registration of non classical mutagenic effects after human irradiation is raised. PMID- 12530151 TI - [Genetic-epidemiologic study of reproductive outcomes in the Ozersk population, located in "Maiak" control area]. AB - In the population of Ozyorsk town situated in the control area of the nuclear enterprise (Production Association "Mayak") the number of pregnancies and its outcomes was studied in cohort of 2.258 women of completed fertility. The mean number of pregnancies per woman is 5.6 +/- 0.07. The mean size of family is 1.9 +/- 0.02. It was estimated that 2.7% of women had no pregnancies, 4.0% of women had no delivery till the end of their reproductive performance. 4.65% of women contributed nothing to the genofond of the next generation, i.e., they had not any children lived till their reproductive performance (20 years). The Crow Index of Opportunity for Selection and its components connected with differential fertility and differential mortality were estimated. In the population under study tow components of selection--selection at the prenatal stages (72.6%) and selection associated with infertility--are shown to be significant. PMID- 12530152 TI - [Predisposition to multifactorial pathology in residents of the city in the zone around the "Maiak" atomic industry enterprise]. AB - According to present achievements in radiation and molecular genetics it is possible to expect that due to chronic radiation exposure the relative increasing of genetic risk in following generations will be observed. It will be due, in the first place, to increasing a percent of multifactorial diseases (MFD). Most of geneticists refer bronchial asthma (BA) to multifactorial diseases. Genetics and epidemiological analysis of liability to BA in population of Ozyorsk situated in the control area of "Mayak" Production Association (PA "Mayak") was accomplished. Population risk, as probability for an individual in population to develop BA up to the end of life is 2.69%. It's higher than in Moscow (2.03%) (p < 0.05). The excess is due to significantly higher values in females (2.96% against 1.93%). Population risk to fall ill of BA in females of Ozyorsk (2.96%) is significantly higher than in males (2.4%). Prevalence of BA in Ozyorsk (0.60%) is significantly higher than in Moscow (0.47%) due to higher values in females (0.65% against 0.47%). The total risk to fall ill of BA amongst relatives is 5.4% that is twice higher than the population risk. The heritability of BA is 0.71 (in Moscow-0.70). PMID- 12530153 TI - [Professional chronic radiation exposure: prospects for studying mutagenic effects in generations using the Southern Ural DNA bank of irradiated people and their offspring]. AB - Data on the established DNA Bank were summarized. The DNA Bank included workers of the Mayak nuclear facility in the Southern Ural, who were exposed to chronic radiation predominantly from external gamma-rays in different doses, and their families for the future study on radiation mutations in somatic cells of parents and possible transmission of genome instability through the germline. At present the DNA Bank contains genetic material from 1500 individuals, among whom there are 223 families. PMID- 12530154 TI - [Cytogenetic results of irradiation of Southern Ural residents]. AB - The paper summarizes the results of studies of 85 individuals exposed in the Southern Ural region. The spontaneous frequency of the cells with micronuclei (MN) in a population of human blood lymphocytes after PHA stimulation and cytokinetic block with cytochalasin B has been determined. The sensitivity of lymphocytes to the irradiation at the dose of 1.0 Gy and the adaptive response (AR) after the irradiation at the low adaptive dose of 0.05 Gy, and the challenge dose of 1.0 Gy 5 h later have been studied too. It was shown that the peculiarity of the Urals population consists in a higher individual variability of the frequency of cells with MN in all groups have been investigated (spontaneous, after acute irradiation in the dose 1.0 Gy) in comparison with Moscow people. The proportion of persons with a significant AR in the Urals groups was considerably lower than that identified among Moscow residents, and the number of persons with enhanced radiosensitivity increased following low-dose irradiation. We can suppose that prolonged action of low level radiation with another ecological factors, living in the contaminated regions result in the enhancement of the sensitivity to the genotoxic agents in the separate individuals. PMID- 12530155 TI - [Long-term cytogenetic effects in children prenatally-exposed to radiation as a result of the accident at the Chernobyl Atomic Energy Station]. AB - Forty-two children exposed to ionizing radiation in prenatal period and 15 children of control group were examined in the remote terms after the accident using the method of differential G-staining of chromosomes in lymphocytes of peripheral blood. It was found that the average group rate of aberrant cells and chromosome aberrations was reliably higher in the children exposed in utero compared to control. Long-term cytogenetic consequences of the pre-natal exposure were characterized by prevalence of aberrations of a chromosome type, mainly stable chromosome lesions. At chronic exposure to low doses of ionizing radiation the increase in the rate both stable and unstable chromosome aberrations. PMID- 12530157 TI - [Cytogenetic effects of alpha-irradiation due to incorporated 239Pu]. AB - Intravenous injection of plutonium dioxide with 1-2 microns particle sizes in amount of 92.5, 46.3 and 23.2 kBq/kg of body mass increased the yield of chromosome aberrations in bone marrow cells of rats by 3.7, 2.3 and 1.7 times, correspondingly, in comparison with the spontaneous level. The model of chromosome aberration dependence on dose of radionuclide was developed based on the experimental results. PMID- 12530156 TI - [Level of chromosome aberrations in peripheral blood lymphocytes in evacuees from the 30-kilometer zone of the Chernobyl Atomic Energy Station and residents of radioactively-polluted territories in the long term after the Chernobyl accident]. AB - Eighteen Ukrainian evacuees from the Chernobyl exclusive zone, twenty one inhabitants of radioactively contaminated areas of Belarus and twelve control donors age-matched to the exposed persons were investigated 14-15 years after the Chernobyl accident for chromosomal aberration yields detected in blood lymphocytes by fluorescence in situ hybridisation technique. Unstable aberration yields measured in both Chernobyl cohorts were close to the background frequencies. Positive age-dependence trends in control donors were determined for the all type stable aberration levels. In evacuees the tendency for diminishing the difference between them and controls for stable aberration levels with persons' age increasing was found. The total stable chromosome exchange yields in evacuees 46-55 years old and inhabitants of areas with low contamination level didn't exceed the control values, but for younger evacuees and inhabitants of sufficiently contaminated regions the statistical increase above the age relevant background meanings was detected for this end-point. The advantages of using the FISH-detectable stable aberrations and particularly the total level of stable chromosome exchanges as the end-points for retrospective biological indication of past radiation exposure in Chernobyl cohorts were discussed. PMID- 12530159 TI - [Assessment of the status of biological health of the population of tundra Nentsy inhabiting northwestern Siberia from data on analysis of autoantibodies and lipoproteins from blood donors]. AB - As a result of large-scale nuclear explosion on Novaya Zemlya test site (1955-62) the Tundra Nentsy population of Yamal-Nentsy autonomous region (YNAR) fell under constant influence of incorporated radioactive isotopes (137Cs and 90Sr). Therefore, it is very important to analyze a possible spectrum of diseases of Tundra Nentsy population. We have developed recently a new method for determination of concentrations of all main fraction and subfraction of lipoproteins (LP, 30 parameters) in human sera using small-angle X-ray scattering, and a general mathematical model to describe LP composition in human blood. The analysis of the 30 parameters characterizing fine spectrum of LPs in 374 YNAR natives showed that only approximately 10% of the donors are normal, while the indices for approximately 90% of the test subjects fall into the range of different pathologies (3-8% incidence in normal population, according to epidemiological studies). Moreover, we found that approximately 41% of European and approximately 56% of Tundra Nentsy have high level of autoantibodies to DNA and cardiolopine like the same for autoimmune diseases patients. PMID- 12530158 TI - [Induced leukemias and their connection with radiation exposure]. AB - The cytogenetic study of bone marrow cells was performed in 40 patients with secondary leukemias which have arisen after application of cytostatic and/or radiotherapy for primary tumours (Hodgkin's disease, lymphomas, acute lymphoblastic leukemias, breast cancer and other solid tumours). The comparative analysis of results has shown, that the leukemias after irradiating or application of alkylating agents and irradiation, have the quite particular clinico-morphological and cytogenetic characteristics. In 70% of cases these diseases develops as smouldering leukemias with subsequent transformation in M-4, M-6, and rarely M-2 cytochemical variants. Primary cytogenetic events in 60% of researched karyotypes are the losses of long arms or whole chromosomes 5 and 7. In 20% of the researched cases normal karyotype was found, in the left 20%, the changes of a karyotype not including anomalies 5 and 7 chromosomes were detected. The obtained outcomes allow to consider the discharged complex of tags as reference for leukemias, induced by irradiating or chemical agents with similar mechanism of action (alkylating agents, benzene and its derivates). This complex of tags is typical for induced leukemias, and in a combination with definition of a level of stable aberrations in peripheral blood lymphocytes, can be utilised for abjection of radiation-induced leukemias from common mass of cases detected in regions, polluted by radionuclides. In this study in 60% of cases only specific for secondary leukemias chromosomal aberrations, including monosomy 5 and 7, rearrangements of 11q23 were found. On the base of the obtained data the differences in concepts of "secondary" and "induced" leukemias are considered. PMID- 12530160 TI - [Dynamic study of the cytogenetic effects in blood lymphocytes from humans living in the Chernobyl Atomic Energy Station exclusion zone without permission]. AB - The cytogenetic examination of a group of people, self-residing in ChNPP Exclusion Zone with soil contamination density 137Cs 74-477 kBq/m2, 90Sr 33-289 kBq/m2, 238, 239 + 240Pu 1.5-10.0 kBq/m2, was conducted in 1998-1999 and also 2001. It is shown, that after 12-15 years of the accident the level of chromosome damages in Zone self-residents' lymphocytes detected by a routine analysis is higher then at the residents of control Yagotin district, Kiev region and comes to a plateau. Lymphocytes with multiple chromosome damages are detected. The probability of its transuranium elements induction which are present in the Zone is discussed at present. PMID- 12530161 TI - [Expression of genomic instability in lymphocytes from children living under conditions of long term exposure to radioactive factors]. AB - An experimental study of expression of genomic instability was carried out with the use of testing irradiation in peripheral blood lymphocytes of 15 children (born in 1986-1998) living on territories with radionuclide contaminations (over 15 Ci/km2, 137Cs, Novozybkov district, Bryansk region). In 5 children exposed to intrauterine irradiation in 1986 this phenomenon was studied in 3 successive cell generations (mitoses). The data obtained suggest the reality of expression of induced genomic instability in the offsprings of repeatedly divided cells of a growing organism exposed to prolonged action of low radiation levels. PMID- 12530162 TI - [Restoration of pine plantations after effect of ionizing radiation in the region of the accident at the Chernobyl Atomic Energy Station]. AB - The main results of the 12-year radiation-genetic monitoring of radiobiological, cytogenetic, and genetic parameters in the Pinus sylvestris L. forest plantations from the zone of the accident at the Chernobyl nuclear power plant presented. Acute ionizing irradiation at doses > 1 Gy was shown to induce the formation of morphoses and depressed growth; at doses > 2 Gy, the reproductive ability of the trees declined. The radiobiological parameters showed a linear (or close to linear) dose-effect relationship. Acute irradiation at a dose of 0.5 Gy induced cytogenetic and genetic effects that were significantly higher than the corresponding control values. The relationship between the cytogenetic effects and the absorbed dose was exponential. The dependence of the mutation frequency at specific loci on the absorbed dose was described by a nonlinear curve. The results of cytogenetic analysis of sprouts obtained from seeds annually (1986 1998) collected in zones of slight, moderate, and strong damage of Pinus sylvestris L. are presented. PMID- 12530163 TI - [Cytogenetic effects in plant populations from the East Urals Radioactive Track]. AB - Mutation dynamics in generations was studied in natural populations of plants from the East Ural Radioactive Track (EURT, Kyshtym accident). The experiments were aimed at the investigation of the formation of genetic load in natural plant populations under conditions of prolonged radiation exposure. The main approach used in the study was the analysis of cytogenetic alteration, chlorophyll and gene mutations in plants exposed for more than 40 years to chronic beta irradiation and in their progeny. PMID- 12530164 TI - Problems of the complex assessment of radiobioecological situation and public health in uranium-extraction regions of Kazakhstan. AB - In the article was considered main direction in investigation and methodological approach in the estimations of radiobioecological situation and condition health of population in the uranium-extraction regions of Kazakhstan. Elimination of consequences of uranium extraction in Kazakhstan is only possible with a broad scientific cooperation and combined resources and man-power from Kazakhstan, from former Soviet Union countries and from other countries. PMID- 12530165 TI - [Effect of extreme factors, differing in nature, on distribution of repetitive sequences in extracellular DNA]. AB - Extreme exposures induce changes of content and fractional composition of extracellular DNA. The degree of these changes is dependent on the nature of stressor and conditioned by genome destabilization. The character of distribution in the separate fractions instable in genome of repeating sequences determines the function of extracellular DNA as an adaptogenic factor. PMID- 12530166 TI - [Study of reparative DNA synthesis in lymphocytes of persons occupationally exposed to radiation]. AB - The UDS efficiency in lymphocytes of professionals chronically exposed to gamma/neutron radiation, as well as for a control cohort was estimated. A credible reduction of UV-induced UDS (KUV) index as compared to the control was demonstrated. This shows an invalid repair state of blood cells in professionals. As for the control cohort, the decreasing tendency of UDS index with age was found. The correlative analysis of UDS index dependence upon an absorbed dose (based on physical dosimetry data) revealed the trend towards repair index reduction along with a higher total absorbed doze, that is followed by UDS index coming onto a plateau. It was also demonstrated that after a sharp and/or accidental irradiation UDS index reduces as compared to permanent portioned irradiation. It was not observed the influence of smoking upon UDS efficiency in blood lymphocytes neither for control, nor for experimental group. The analysis of vitamin therapy of the both cohorts showed that such therapy raises UDS index of the control (i.e., non-professional) group. Besides, it was found out an additional positive effect of vitamin therapy, i.e. doze leveling in long-term perspective after irradiation. The most expressed KUV/doze correlation was characteristic for professionals, who do not take vitamins and do not smoke. This proves that in blood cells of professionals it is preserved a dependence of repair system invalidity upon the absorbed doze without similar external factors. PMID- 12530167 TI - [Effect of space flight factors on health of cosmonauts in the near and late term after space flights]. AB - Cytogenetical studies of cosmonauts' peripheral blood lymphocytes after space flights on MIR orbital station showed a statistically significant increase in the yields of radiation-induced chromosomal aberrations. However, similar studies with in vitro irradiation of biological objects with accelerated charged particles are of great importance for elucidation of the nature of cytogenetical damage induced in vivo. It is also important to investigate the structure of cosmonatus' diseases over their life, in particular, lens opacities and oncological diseases. Thus, the purpose of the investigations planned is to study cytogenetical damage in blood lymphocytes from cosmonauts after space flights on the ISS in vivo, as well as in donor blood lymphocytes after in vitro exposure to accelerated charged particles. The tasks of the project are as follows: determination of the yields and types of chromosomal aberrations in cosmonauts' blood lymphocytes before and after space flights, comparative studies of biological effects induced in vitro by different types of ionizing radiation in human blood lymphocytes in ground experiments, assessment of cytogenetical risks, analysis of the structure of cosmonatus' diseases comparing with that of whole population, study of the mortality and frequency of cataracts and oncological diseases in cosmonauts. The results to be obtained will be used for setting of health norms applied to the influence of radiations of different types, and for elaboration of measures to reduce health risks from space flight factors. PMID- 12530168 TI - [Osteoporosis in premenopausal women]. AB - Osteoporosis is a systemic disease of bone, which is characterized by decreased bone mass and changes in the microarchitecture of bone tissue followed by brittleness of bones and increased risk of fractures. Osteoporosis frequently is a disease of postmenopausal women, nevertheless, in rare cases, osteoporosis can also occur in young adults. There are only few studies on the pathophysiology of "premenopausal osteoporosis"; in addition to idiopathic forms, osteoporosis in young women can be caused by glucocorticoid treatment, by eating disorders or can be associated with pregnancy. PMID- 12530169 TI - Differences in distribution of T-scores and Z-scores among bone densitometry tests in postmenopausal women (a comparative study). AB - BACKGROUND: To determine the character of T-score and Z-score value distribution in individually selected methods of bone densitometry and to compare them using statistical analysis. METHODS: We examined 56 postmenopausal women with an age between 43 and 68 years with osteopenia or osteoporosis according to the WHO classification. The following measurements were made in each patient: T-score and Z-score for: 1) Stiffness index (S) of the left heel bone, USM (index). 2) Bone mineral density of the left heel bone (BMDh), DEXA (g of Ca hydroxyapatite per cm2). 3) Bone mineral density of trabecular bone of the L1 vertebra (BMDL1), QCT (mg of Ca hydroxyapatite per cm3). The densitometers used in the study were: Ultrasonometer to measure heel bone, Achilles plus LUNAR, USA; DEXA to measure heel bone, PIXI, LUNAR, USA; QCT to measure the L1 vertebra, CT, SOMATOM Plus, Siemens, Germany. STATISTICAL ANALYSIS: Differences between measured values of T scores (Z-scores) were evaluated by parametric or non-parametric methods of determining the 95% confidence intervals (C.I.) RESULTS: Differences between Z score and T-score values for compared measurements were statistically significant; however, these differences were lower for Z-scores. CONCLUSIONS: Largest differences in 95% C.I., characterizing individual measurements of T score values (in comparison with Z-scores), were found for those densitometers whose age range of the reference groups of young adults differed the most, and conversely, the smallest differences in T-score values were found when the differences between the age ranges of reference groups were smallest. The higher variation in T-score values in comparison to Z-scores is also caused by a non standard selection of the reference groups of young adults for the QCT, PIXI and Achilles Plus densitometers used in the study. Age characteristics of the reference group for T-scores should be standardized for all types of densitometers. PMID- 12530170 TI - [Guidelines for drug therapy of postmenopausal osteoporosis]. AB - Osteoporosis is known as a condition characterized by low bone mass and microarchitectural deterioration of bone tissue leading to bone fragility and a consequent susceptibility to fracture. Osteoporosis has its highest rate of occurrence in postmenopausal women, and in Western countries it has been estimated that for white women aged fifty, the life-time risk of developing an osteoporotic fracture is nearly 40%. Given the consequences of osteoporosis, the most important goal of therapy is to prevent fractures. In Austria, several pharmacologic options for treatment of osteoporosis are available, including bisphosphonates (alendronate, etidronate, risedronate), selective estrogen receptor modulators (raloxifene), calcitonins (salm-calcitonin, elcatonin), fluorides (sodium-fluoride, monofluorophosphate), anabolic steroids (nandrolone decanoate), steroid derivates (tibolone), estrogen and hormone replacement therapy. An evidence-based evaluation of these treatment options clearly indicates that alendronate, risedronate and raloxifene sufficiently reduce the risk of vertebral fractures. There is less evidence for reduction of vertebral fracture risk for etidronate, calcitonin, estrogen replacement therapy or hormone replacement therapy. Only alendronate and risedronate have been shown to reduce the risk of hip fractures. Calcium and vitamin D are useful adjuncts to any specific treatment for osteoporosis, particularly when Calcium and vitamin D deficiencies have been diagnosed. Also, there is good evidence that in women with Calcium and vitamin D deficiency, a combination of Calcium and vitamin D may reduce the risk of non-vertebral fractures. There is no evidence so far that a combination therapy of antiresorptive drugs would result in reduced fracture risk. PMID- 12530172 TI - [So-called morbidity]. AB - True statistical data concerning human minimal health-disturbances--even of single persons--are difficult to get. It is unlikely that we ever will have broad knowledge about these minimal deviations from health and the border between pathology and physiology. However this region was opened up by us to future research. We propose to give these occurrences the name 'Minimity'. PMID- 12530171 TI - [Etiology and diagnosis of otalgia]. AB - Otalgia is the sensation of pain in the ear, while referred otalgia is pain felt in the ear but originating from a nonotologic source. Ear pain is a diagnostic problem when examination of the ear shows no pathology. Pain in the ear can be caused by inflammation of the external meatus or the middle ear. Nonotogenic otalgia may be due to referred or reflex pain, neuralgia, or to a psychogenic problem. Otalgia may be referred also from primary carcinoma of the head and neck. A thorough understanding of the anatomy of the head and neck is required. Referred pain is due to irritative lesions involving the fifth, ninth, or tenth cranial nerves and spinal nerves C2 and C3. If pathologic problems in the area supplied by these nerves have been ruled out, the neuralgia should be considered in the differential diagnosis. Psychogenic factors must be identified and treated before any type of surgical therapy is recommended. A basic knowledge of neuroanatomy, with emphasis on the cranial nerves, their course, distribution, and function, is also necessary. The complexity of this field may sometimes require the treating physician to call for consultations from colleagues in the fields of oral surgery, neurology, or neurosurgery. The therapy depends of the causes of the ear-ache. PMID- 12530173 TI - [Renaissance of general practice]. AB - Special research work has taught that general practice (family medicine) is a specialization of its own. It requires specific education and vocational training. As far as universities and administrative bodies have accepted it no doctor can start practising family medicine unless he has passed a vocational training of many years duration. That fact together with a successful continued research concerning applied general practice should initiate a true renaissance of general practice. PMID- 12530174 TI - Degree of threat to the biological diversity in the Ilha Grande State Park (RJ) and guidelines for conservation. AB - The State Park of Ilha Grande is only a part (5,594 hectares) of the entire island (19,300 hectares) which is located off the south coast of Rio de Janeiro state, between the cities of Mangaratiba and Angra dos Reis. Approximately half of the Park area (47%) is covered by dense Atlantic forest. The secondary forest growth is in a process of ecological succession close to attaining maturity (43%) and the remaining part (10%) is composed of human-altered areas (1%), rocky outcrops with herbaceous vegetation (7%), mangroves and beaches (2%). The fauna is well represented but already shows signs of degradation with introduced species. The analysis of the degree of threat has shown that the dense forest habitat has a relatively stable status of conservation while the secondary forest, the mangrove and the herbaceous vegetation on rocky outcrops (and their fauna) are categorized as vulnerable. The area altered by human occupation is considered threatened. Since the coastal area where Ilha Grande is located is well known for its beautiful scenery (known as the green coast, because of the contrast between the ocean and the Atlantic forest covering the Serra do Mar mountain chain). There is a strong possibility for tourism to become the means in which to achieve economic sustainability for conservation. Contradictorily, tourism is also the major threat to local biodiversity and its landscape units. Because tourism is not organized and controlled, during high season the numbers grow above local capacity, giving rise to a proliferation of hotels, guesthouses and camping grounds. The resulting untreated open sewage, random garbage disposal and other harmful activities form the major threats to biodiversity. PMID- 12530175 TI - Methodologies for conservation assessments of the genetic biodiversity of aquatic macro-organisms. AB - International organizations and biodiversity scientists recognize three levels of biodiversity: genetic, species, and ecosystem. However, most studies with the goal of assessing biodiversity collect data at only a single level--that of the species. Even when multiple levels of biodiversity are considered, usually only ecosystem diversity is also evaluated. Genetic diversity is virtually never considered. Yet, genetic diversity is essential for the maintenance of populations and species over ecological and evolutionary time periods. Moreover, because components of genetic diversity are independent of either species or ecosystem diversity, genetic diversity can provide a unique measure by which to assess the value of regions for conservation. Regions can be valuable for conservation of their genetic resources regardless of their levels of species or ecosystem uniqueness or diversity. In general, the same methods and statistical programs that are used to answer questions about population genetics and phylogenetics are applicable to conservation genetics. Thus, numerous genetic techniques, laboratory methods, and statistical programs are available for assessing regional levels of genetic diversity for conservation considerations. Here, we provide the rationale, techniques available, field and laboratory protocols, and statistical programs that can be used to estimate the magnitude and type of genetic diversity in regions. We also provide information on how to obtain commonly utilized statistical programs and the type of analyses that they include. The guide that we present here can be used to conduct investigations of the genetic diversity of regions under consideration for conservation of their natural resources. PMID- 12530176 TI - Spatial distribution of Plecoptera nymphs in streams of a mountainous area of Central Brazil. AB - In this paper the spatial distribution of Plecoptera nymphs in the Almas' River basin, Pirenopolis, GO, was studied. Two Surber samples, each comprising 20 sampling units and totalling 2 m2, were taken in each of the 13 stations, one during the rainy season (January 1994) and the second during the dry season (July 1994). In 5 of these stations, monthly samplings were made from June 1993 to July 1994; in these, temperature, velocity, discharge, electrical conductivity and pH were measured. Regional rainfall was also obtained. To ascertain the distribution of nymphs in the habitat, a separate sample was taken. Of the factors considered, the most important affecting the spatial distribution of the stonefly nymphs were altitude, stream order, and anthropic influence. Locally, the genera Anacroneuria and Kempnyia showed clumped distributions, but the data for Gripopteryx and Tupiperla were inconclusive due to low numbers. PMID- 12530177 TI - Spatial use of rodents (Rodentia: Mammalia) host body surface by ectoparasites. AB - We studied the ectoparasite and the Amblyopinini beetle fauna associated with four small mammal species of the Atlantic Rainforest of Ilha Grande, an island located off the southern Rio de Janeiro State Coast, Southeastern Brazil, analyzing to what extent the parasites were specific to each region of the host body. During the study, a total of 90 individual rodents were captured: 61 Proechimys iheringi Thomas, 1911 (Echymyidae), 22 Sciurus aestuans (Thomas, 1901) (Sciuridae), 4 Oxymycterus sp. (Waterhouse, 1837), and 2 Nectomys squamipes (Brants, 1827) (Sigmodontinae). The data showed that the ectoparasites and Amblyopinini on some rodent hosts in Ilha Grande tend to prefer particular host body sites, and that some ectoparasite species sites may overlap owing to their inaccessibility to the host. PMID- 12530178 TI - Effect of seed mass on germination and growth in three dominant species in southern Brazilian coastal dunes. AB - The effect of seed mass on germination and growth was tested in fresh-seeds of Blutaparon portulacoides, Panicum racemosum, and Spartina ciliata, selected at random in southern Brazilian populations. The seed mass varied within a population of the three species. Both B. portulacoides and P. racemosum showed normal frequency distribution of seed mass, while S. ciliata did not. Significant differences were observed in seed germination between large and small seeds of all species. In all species the capacity of seedling elongation was greater in seedlings of large seeds than those of small ones. Relative growth rate of seedlings of P. racemosum and S. ciliata decreased with time in all seed mass size-classes. On the other hand, the relative growth rate of B. portulacoides seedlings increased during the first 40 days. Seed mass is an important biological factor, affecting seed germination, seedling elongation, and growth of these species, and favoring large seeds, specially in areas of active sand accretion like coastal dunes. PMID- 12530179 TI - Rapoport effect in South American Carnivora (Mammalia): null models under geometric and phylogenetic constraints. AB - Rapoport effect predicts that species geographic range sizes will increase toward higher latitudes, probably reflecting adaptations to extreme climatic conditions that increase species tolerance. Recently, studies about spatial patterns in species richness and geographic range size may be associated with the geometry of species' ranges. In this context, null models can be used to search for the causal mechanisms associated with these patterns. In this paper, we analyzed Rapoport effect using a null model to evaluate how phylogenetic structure and geometric constraints simultaneously affect latitudinal extents of 40 species of South American terrestrial Carnivora. The latitudinal extents of Carnivora tended to decrease toward Southern latitudes, in the opposite direction expected under a simple Rapoport effect, but in accordance to geometric expectations of position of midpoints in the continent. Using 5000 simulations, it was possible to show that the null regression coefficients of latitudinal extents against midpoints are positively biased, reflecting the geometric constraints in the latitudinal extents. The results were equivalent in phylogenetic and non-phylogenetic analyses. The observed regression coefficient was significantly smaller (line is less inclined) than expected by chance alone, demonstrating that the geometric constraints in the latitudinal extents exist even after controlling for phylogenetic structure in data using eigenvector regressions. This suggests that the "spirit" of Rapoport effect (sensu Lyons & Willig, 1997) could be maintained, i.e., that latitudinal extents in Southern region of the continent are relatively larger than those in Northern regions, even after controlling for phylogenetic effects. PMID- 12530180 TI - On the natural diet of Daphnia laevis in the eutrophic Pampulha reservoir (Belo Horizonte, Minas Gerais). AB - The aim of this study was to assess the major food items ingested by adult specimens of Daphnia laevis within the eutrophic Pampulha reservoir in Belo Horizonte, Minas Gerais, Brazil. The gut content was analyzed after addition of sodium hypochlorite and also through the examination of dissected guts under scanning electron microscopy. The results showed that Chlorophyceae was the main food item ingested, representing c. 80.5% of the total ingested food. Moreover, Eutetramorus fottii, Coelastrum pseudomicroporum and Oocystis lacustris, the dominant phytoplankton species within the reservoir, were the most frequent cells found in the gut contents. Euglenophyta also represented an important food item accounting for 15% of the ingested material, including mainly Trachelomonas volvocina and Euglena oxyuris, although less abundant in the reservoir (< 10% of total phytoplankton). Blue-green algae occurred at much lower percentages in the guts than in the phytoplankton. A small amount of undigested Microcystis aeruginosa colonies were also found in the gut content of D. laevis. Scanning electron microscopy results showed that, besides phytoplankton cells, a great amount of abiogenic material was also ingested. The amount of this inorganic material increased considerably in the tract (from 15% to 75% of the gut content), when a peak of D. laevis was observed in the reservoir. Our assumption is that the ingestion of this inorganic material can be a strategy used by D. laevis to obtain additional food supply. PMID- 12530181 TI - Some Digenea parasites of tunny from the coast of Rio de Janeiro State, Brazil. AB - Rhipidocotyle pentagonum (Ozaki, 1924) is reported for the first time in South America parasitizing Auxis thazard and in a new host Katsuwonus pelamis. Tergestia laticollis (Rudolphi, 1819) is reported for the first time in South America and in Thunnus albacares, representing a new host record. Copiatestes filiferus (Leuckart, in Sars, 1885) is recorded for the first time in Brazil and in Thunnus albacares, another new host record. Tetrochetus coryphaenae (Yamaguti, 1934) is presented for the first time in Brazil parasitizing Thunnus albacares. PMID- 12530182 TI - Report on two species of Digenea from marine fishes in Brazil. AB - Two species of Digenea were recorded for the first time in South America and in new hosts: Acanthostomum spiniceps (Looss, 1896) (Cryptogonimidae) was reported from Astroscopus sexspinosus (Steindachner, 1877) (Uranoscopidae) and Diplomonorchis sphaerovarium Nahhas & Cable, 1964 (Monorchiidae) from Ophichthus gomesi (Castelnau, 1855) (Ophichthidae). From the latter, Heliconema heliconema Travassos, 1919 (Nematoda, Physalopteridae), was also recovered representing also a new host for this nematode species. PMID- 12530183 TI - On two methods to estimate production of Eichhornia crassipes in the eutrophic Pampulha reservoir (MG, Brazil). AB - This paper deals with variations in biomass and in the production of Eichhornia crassipes (water hyacinth) in a eutrophic tropical reservoir. Net production was assessed by both a traditional method and a demographic approach. The initial biomass ranges from 96 g DW.m-2 to a maximum of 2027 g DW.m-2. Net production estimates varied greatly according to which method was used to calculate them (traditional: 20.6 ton DW.hectare-1.year-1, demographic: 226.9 ton DW.year-1). PMID- 12530184 TI - Fimicolous Histeridae coleoptera in Campo Grande, MS, Brazil. AB - Fecal masses recently excreted and/or almost dry were collected weekly in a pasture of Brachiaria decumbens Stapf, from May 1990 to April 1992. The feces were conditioned in 15-liter opaque plastic buckets, containing lateral and top openings, where flasks were fastened for capturing Histeridae beetles present in these masses. Three thousand two hundred ninety-nine specimens were collected belonging to 11 species in the Genus: Phelister, Hister, Euspilotus, Acritus, and Xerosaprinus. The most frequent, constant, and abundant species were Phelister sp. nr. carinifrons and P. haemorrhous. PMID- 12530185 TI - Occurrence and biogeographic aspects of Melipona quinquefasciata in NE Brazil (Hymenoptera, Apidae). AB - The stingless bee Melipona quinquefasciata is not included among the nine bee species of Melipona described in literature of NE Brazil. However, reports of some farmers raised suspicion on the occurrence of M. quinquefasciata in the state of Ceara, in NE Brazil. Investigations were carried out from July 1997 to September 2000, by means of trips to the areas of probable occurrence of this bee species. Results confirmed the presence of M. quinquefasciata in Ceara and determined its habitat along the chapada do Araripe (Araripe plateau) and all extension of planalto da Ibiapaba (Ibiapaba plateau), in altitudes between 600 and 900 m. Melipona quinquefasciata lives in the phytocoenosis of cerrado (Brazilian savanna), cerradao (savanna forest) and carrasco (montane deciduous shrub vegetation) on the top of Araripe plateau, and only carrasco in the Ibiapaba plateau. Due to pressures caused by reduction of the area covered with native vegetation, large use of agrochemicals in anthropic areas and generalised predatory hunting of honey and beeswax, M. quinquefasciata is in risk of disappearing from the ecosystems of Araripe and Ibiapaba plateaus within a few years. PMID- 12530186 TI - Reproductive biology of a Characidiinae (Osteichthyes, Characidae) from the Ubatiba River, Marica-RJ. AB - The Ubatiba river is a coastal stream subject to stochastic spates all over the year and, Characidium sp.n. is among the twenty-two species that compose its ichthyofauna. In this study we analyse some traits of its reproductive biology and discuss whether the adopted strategy has causal relationship with the environmental variables. Specimens were collected in the upper Ubatiba river. Samplings, were carried out monthly by electrofishing, between Oct./94 and Sep./95. Some differences between male and female strategies were observed. Females were significantly (p < 0.05) bigger than males; length--weight relationship was different (p < 0.05) between sexes with the onset of sexual maturity occuring at smaller sizes among males. Reproductive investment is high for both males and females but higher for males, maximum Gonadosomatic Index for females and males were 40.97% and 44.90%, respectively. Reproductive specimens were registered all over the year, suggesting continuous reproduction. High values of fecundity were also registered varying from 1342 to 5535 eggs for ripe females of 4.4 and 7.0 cm, respectively and an amount of 1105 oocytes per grams of fish. We suggest that differences in the reproductive strategy, between sexes, determine the observed patterns in the size structure; the absence of males in the higher SL classes could be a consequence of high mortality rates and/or reduction in the growth rates as a precocity consequence in the onset of reproduction and/or high reproductive investment. Relationship between continuous reproduction and environmental condition could be explained as an adaptation to maximise supervivency of the young fishes that is: guarantee the species maintenance in a stochastic environment. PMID- 12530187 TI - Specialized metabolism and biochemical suppression during aestivation of the extant South American lungfish--Lepidosiren paradoxa. AB - Lepidosiren paradoxa (piramboia) is the single representative of Dipnoan (lungfish) in South America. This species is considered a living fossil, in spite of some reports describing this fish as having a very specialized life style. It aestivates during the dry season, and has developed metabolic adaptations to cope with both flooding and drought. The literature describing its tissue ultra structure shows high glycogen stored in the muscle, suggesting a strong dependence on anaerobic glycolysis. The present paper reports tissue enzyme levels of LDH, MDH, and CS, and isozymic tissue distribution of LDH, MDH, ADH, PGI, SOD, and PGM of 7 aestivating specimens from Lago do Canteiro in the Amazonas River. Animals were caught while burrowed in mud during the aestivation period. Our findings reveal high anaerobic capacity of both skeletal and heart muscles, even during the aestivation period, when enzymes showed suppressed levels compared to those of non-aestivating animals (data from the literature). Isozymic patterns suggest loss of duplicate condition in most analyzed loci, a characteristic that occurs mainly in higher vertebrate categories. These data indicate that, compared to the fish group, lungfish may be considered advanced, despite retaining primitive morphological characters. PMID- 12530188 TI - Genetic diversity in different populations of sloths assessed by DNA fingerprinting. AB - In this study we analyzed a population of Bradypus torquatus with individuals originally distributed in different localities of Bahia, and two populations of B. variegatus with individuals from Bahia and Sao Paulo States. Using the DNA fingerprinting method, we assessed the genetic variability within and between populations. Analysis of the DNA profiles revealed genetic similarity indices ranging from 0.34 +/- 0.07 to 0.87 +/- 0.04. Similar low levels of genetic variability were found only in isolated mammalian populations or among related individuals. This study presents the first analyses of genetic diversity in sloth populations. PMID- 12530189 TI - Rotifer production in a shallow artificial lake (Lobo-Broa Reservoir, SP, Brazil). AB - Rotifer production in a shallow artificial lake (Lobo-Broa Reservoir) was studied for twenty consecutive days between August 25, 1995 and September 13, 1995, during the dry season. Two species were dominant, during this period namely Filinia pejleri and Keratella americana, contributing, respectively, 36% and 28% to total rotifer numbers. Production estimates of these species were, respectively, 41.93 and 80.70 micrograms dry weight.m-3.d-1, corresponding to 18.45 and 35.51 micrograms Cm-3.d-1. The population of F. pejleri was numerically more abundant than that of K. americana, while the production of the former was less, as a result of lower egg to female ratios. PMID- 12530190 TI - The effects of treatment with melatonin on the ultrastructure of mouse Leydig cells: a quantitative study. AB - Both the presence of receptors for gonadal steroids in the pineal gland and in vitro observations of direct action of melatonin upon Leydig cells, inhibiting testosterone secretion, indicate a direct connection between pineal gland and gonadal function. In the present study, we used a transmission electron microscope to analyze the morphologic parameters of Leydig cells from adult Swiss outbred white mice treated with daily subcutaneous injections of 100 micrograms of melatonin (N-acetyl, 5-methoxytryptamine), during 22 consecutive days, compared with sham-control animals which had only received the melatonin vehicle. The melatonin group of mice showed a decrease in nuclear volume and fractional nuclear volume; smooth and rough endoplasmic reticulum; mitochondria; and Golgi complex. Our data also showed an increase in cytoplasmic volume, fractional cytoplasmic volume, and lysosomes in these same animals. The results suggest that melatonin, directly or indirectly, alters the ultrastructure of mouse Leydig cells and possibly influences their secretory activity by inhibiting their capacity to secrete steroids. PMID- 12530191 TI - Composition and abundance of zooplankton in the limnetic zone of seven reservoirs of the Paranapanema River, Brazil. AB - The species composition and abundance of the zooplankton community of seven reservoirs of the Paranapanema River, located between 22 degrees 37'-23 degrees 11'S and 48 degrees 55'-50 degrees 32'W, were analysed over four periods, in the year of 1979. The zooplankton community was composed of 76 species of Rotifera, 26 species of Cladocera and 7 species of Copepoda. For a large part of the period under study the Rotifera were dominant, followed by Copepoda. The Piraju and Salto Grande reservoirs, which occupy intermediate positions in the cascade of reservoirs, were richest in species, most of them belonging to Rotifera and Cladocera. In the reservoirs Rio Pari and Rio Novo, lateral to the cascade of reservoirs, a lower species richness was observed, although higher densities of organisms were found than in the other reservoirs located in the main river body. Different rotifer species occurred in succession, being abundant in different periods, with no defined pattern. Among the copepods, Thermocyclops decipiens predominated in the majority of the reservoirs. Ceriodaphnia cornuta was the most abundant cladoceran in the intermediate reservoirs of the cascade, and Daphnia gessneri, Bosminopsis deitersi and Moina minuta, in the reservoirs lateral to the cascade. The most frequent zooplankton species were Notodiaptomus conifer, Thermocyclops decipiens, Ceriodaphnia cornuta cornuta and C. cornuta rigaudi, Daphnia gessneri, Bosmina hagmanni, Keratella cochlearis and Polyarthra vulgaris. Some relationships were found between the trophic state of the reservoirs and the zooplankton community. PMID- 12530192 TI - Histological and histochemical aspects of the penial glands of Girardia biapertura Sluys, 1997 (Platyhelminthes, Tricladida, Paludicola). AB - Girardia biapertura was described with sperm ducts penetrating the penis bulb, subsequently opening separately at the tip of the penis papilla and receiving the abundant secretion of penial glands. In the present work, the penial glands of this species have been histologically and histochemically analysed, and four types of secretory cells are distinguished. The openings of the penial glands into the intrabulbar and intrapapillar sperm ducts, designated here as intrapenial ducts, allow for the distinction between three histologically differentiated regions. The most proximal region possibly corresponds to the bulbar cavity of other freshwater triclads whereas the median and distal portions correspond to the ejaculatory duct. The proximal region of the intrapenial ducts receives mainly the openings of a secretory cell type (type I) that produces a proteinaceous secretion. A second type of secretory cell (type II) that secretes neutral mucopolyssacharides opens into the median region of the intrapenial ducts. The distal portion of the ducts receives two types of secretory cells (types III and IV) which secret glycoprotein and glycosaminoglycans, respectively. Types III and IV open also directly into the male atrium through the epithelium of the penis papilla. A comparison with the results presented here and those of other authors for species of Girardia is provided and the importance of the study of the penial glands for taxonomic characterisation of freshwater triclads is emphasised. PMID- 12530193 TI - [Computerized 3-D model to study biomechanics of the middle ear using the finite element method]. AB - Based on known data about the behavior of the sound transmission mechanism, we generate a model that reproduces the real mechanics of the middle ear by the finite elements method in the most exact manner possible. We designed a computerized model to simulate the biomechanic behavior of the ear drum/ossicular chain. The investigation focused on a possible clinical application, given that the variables necessary for simulation of several conditions can be introduced in our model: tympanic perforation, fixation of the ossicular chain, chain interruption, etc. Introducing the characteristics of the lesion virtually in a simulator program market is possible to reach conclusions about the conductive hypoacusis created and about therapeutic possibilities. The 3-D modelling of the tympanic membrane, malleus, incus and stapes is presented, considering form, weight, density, Young's module (N/m2) and Poisson coefficient for the different parts of each ossicle. The modeling of the union of the different tympanic ossicular elements, including muscles and ligaments, is also shown. PMID- 12530194 TI - [Cochlear cytotoxic activity of cisplatin in experimentation animals. A study using scanning electron microscopy]. AB - Cisplatin is an antitumoral agent commonly used in the treatment of different solid tumors. Its use induces adverse effects like oto- and nephrotoxicity due to acute renal failure. This makes it a suitable drug when creating an experimental model of ototoxicity. Our aim is to study the cellular lesions induced in the cochlea of the rat and the guinea pig by variable doses of cisplatin injected intraperitoneally, after short, middle, and long term survival, by means of scanning electronic microscopy. Results show the global effect of the cells of the organ of Corti with a more intense initial damage of the supporting cells, continuing with the late effect of the sensorial cells. PMID- 12530195 TI - [Bioethical issues in pediatric cochlear implants]. AB - The use of cochlear implants can modify significantly the way in which deaf children relate with the outside world through psychological, linguistic and cognitive changes. The main question about this subject is the ethical controversy with regards to who has the right to make such a decision for a young child. We present the children evaluated at our hospital in order to assess the cochlear implant option. That population was of 37 implanted children, 10 children waiting for surgery and 21 rejected children. We describe and analyze from the bioethical bases and methodology the problems found: decision of no implant in children of hearing parents, the deaf culture point of view, the mature minor doctrine, teenagers, social and cultural problems, multihandicapped children and responsibility of the implants team. PMID- 12530196 TI - [Incidence of A1555G mutations in the mitochondrial DNA and 35delG in the GJB2 gene (connexin-26) in families with late onset non-syndromic sensorineural hearing loss from Cantabria]. AB - INTRODUCTION: Sensorineural deafness is a very common disorder in humans, which affects approximately 10% of the population. Genetic causes are suggested to be responsible for more than half of the cases. The A1555G mutation in the mitochondrial 12S rRNA gene and the 35delG mutation in the GJB2 gene are the most common mutations for sensorineural deafness in the Spanish population. METHODS: A genetic study was carried out in order to determine the frequency of the mutations A1555G in the mitochondrial DNA and 35delG in the connexin-26 gene in 21 patients from 21 non-consanguineous unrelated families affected by late-onset bilateral non-syndromic sensorineural hearing loss from Cantabria. RESULTS: The A1555G mutation was found in 6 patients. Five of these 6 patients had been treated with aminoglycosides. In all of them the auditory impairment affected mainly the high frequencies. The 35delG mutation was not found in any of the patients. CONCLUSIONS: The A1555G mutation in the mitochondrial DNA has been found to be the most common amongst the Cantabrian population. The A1555G mutation should be suspected in those members of families affected by sensorineural hearing impairment with a maternal inheritance pattern and ototoxicity from treatment with aminoglycoside antibiotics. The 35delG mutation in the GJB2 gene does not seem to be a major cause of deafness in families with late-onset non-syndromic sensorineural hearing loss in our area. PMID- 12530197 TI - [Risk factors involved in presbyacusis]. AB - INTRODUCTION: Presbyacusis is a disease whose prevalence tends to increase in Europe, due to the aging of the population. Our purpose is to evaluate the prevalence of presbyacusis, as well as the implication of sex, cardiovascular risk factors and relation with home and its geographical location. PATIENTS AND METHOD: We started a cross-sectional study with 61 patients coming from different retirement homes, being able to finally test 59 subjects, after two abandonments. RESULTS: The prevalence of presbyacusis was of 16.9%, predominating in female sex, although it was not a statistically significant association, with an average age at the moment of the study of 75.7 years. None of the presbyacusis patients referred smoking habits in the last 30 years, and only sporadic alcohol. A significantly negative statistical relation with the hypertension was verified. There was no significant statistical association with the location and type of home, neither with other risk factors. CONCLUSIONS: The high prevalence of the disease is confirmed, without a real significant predominance in women. The time of evolution of the disease is variable at the moment of the diagnosis. We emphasized the absence of relation between the arterial hypertension, the consumption of tobacco and alcohol and presbyacusis, as well as the rest of studied risk factors. PMID- 12530198 TI - [Tympanoplasty using cartilage: 3 years of experience]. AB - We present our results in a timpanoplasty technique using autologous cartilage, in a retrospective study of 71 cases within 3 years. We used this technique in revision surgery, large perforations, severe retractions, and atrophic membranes. 32% of cases needed ossicular chain reconstruction. A complete closing of the membrane was obtained in 86% of cases, while 14% remained re-perforated or discontinued between cartilage pieces. These results are comparable to other studies using cartilage graft with several techniques. These results encourage us to continue using this technique in order to acquire more experience and a better knowledge about the efficacy of this technique. PMID- 12530199 TI - [Use of median fronto-naso-orbital flap to decrease the incidence of complications in patients undergoing craniofacial resection]. AB - BACKGROUND: Resection of malignant neoplasms invading the anterior cranial base is performed with craniofacial resection (CFR) with acceptable results in complication rates and oncologic outcomes. However, still there are series with major morbidity in up to 40% of patients, and mortality of 5%. A retrospective study was performed to evaluate the results in terms of morbidity, mortality, function and aesthetics using a median fronto orbital flap (MFOF). METHODS: The MFOF was used in 28 consecutive patients who presented to the Head and Neck Service from 1992 to 1999, in the Instituto Nacional de Cancerologia, Mexico City, and was combined with a superior rhinotomy for en bloc resection of the ethmoid complex in the last 12 cases. 89% of patients had malignant neoplasms. RESULTS: 1 death occurred in this series (3%) and the global morbidity was 35%. Aesthetics and function were good to excellent. Percentages of tumor-free survival for malignant lesions was 76% and 41% at 2 and 5 years follow up, respectively. CONCLUSION: CFR is a safe approach for treatment of neoplasms of the anterior skull base. MFOF mobilization decreases complication rates and gives superb exposure for en bloc resection of tumors invading the ethmoidomaxillary complex specially when combined with a superior rhinotomy. A positive impact on quality of life was obtained. PMID- 12530200 TI - [A study of cervical metastases from unknown primary tumor]. AB - Cervical metastases of cancer of an unknown primary tumour (CMUPT) are infrequent but they represent botha diagnostic and therapeutic challenge for ENT physicians. We present a retrospective study over 22 patients with CMUPT diagnosed in our hospital during 13 years (3.4% of the patients with head and neck cancer). The sensitivity of the fine-needle aspiration technique was 94.7%. 54% were N2a, 36% N3 and 9% N2b. 73% were squamous cell carcinoma and 27% were undifferentiated carcinoma. We established our protocol on diagnosis and management: panendoscopy with biopsies of nasopharynx, vallecula and pyriform sinus and ipsilateral tonsillectomy. The primary source was identified in 23% of the series. All squamous cell carcinoma were treated with radical neck surgery plus radiotherapy and the undifferentiated carcinoma with chemotherapy and radiotherapy. The overall survival was 64% at 3 years. The N-stage affected the survival rate. PMID- 12530201 TI - [Two cases of nasal rhinosporidiosis]. AB - The rhinosporidiosis is an illness caused by Rhinosporidium Seeber, which affects not only the man but also other animals. The usual pattern of manifestations are located and characterized either by the development of big polyps, tumours, papillomas or wart lesions. These lesions are soft and hyperplastic, highly vascular and they can be fixed or pedunculated. Our study reports two cases in young people, from the same geographical area, who showed papillomatoses tumours in the nasal cavity. The diagnoses of rhinosporidiosis nasal was done through a histopathological study. PMID- 12530202 TI - [Neck liposarcoma. Presentation of one case and revision of the literature]. AB - Neck liposarcoma is an infrequent tumor in the head and neck area. A case in a 70 years male is reported with a presentation the in inferior third of the lateral neck region. Two previous surgical procedures and Radiotherapy (50 Gy) were carried out. A modified radical dissection was performed. No metastatic nodes were found. A revision of the literature is done with special remarks on prognostic factors; histology and surgical treatment with wide margins are favourable factors. PMID- 12530203 TI - Pheromone biosynthetic pathways in the moths Helicoverpa zea and Helicoverpa assulta. AB - Sex pheromones of many Lepidopteran species have relatively simple structures consisting of a hydrocarbon chain with a functional group and usually one to several double bonds. The sex pheromones are usually derived from fatty acids through a specific biosynthetic pathway. We investigated the incorporation of deuterium-labeled palmitic and stearic acid precursors into pheromone components of Helicoverpa zea and Helicoverpa assulta. The major pheromone component for H. zea is (Z)11-hexadecenal (Z11-16:Ald) while H. assulta utilizes (Z)9-hexadecenal (Z9-16:Ald). We found that H. zea uses palmitic acid to form Z11-16:Ald via delta 11 desaturation and reduction, but also requires stearic acid to biosynthesize the minor pheromone components Z9-16:Ald and Z7-16:Ald. The Z9-16:Ald is produced by delta 11 desaturation of stearic acid followed by one round of chain shortening and reduction to the aldehyde. The Z7-16:Ald is produced by delta 9 desaturation of stearic acid followed by one round of chain-shortening and reduction to the aldehyde. H. assulta uses palmitic acid as a substrate to form Z9-16:Ald, Z11-16:Ald and 16:Ald. The amount of labeling indicated that the delta 9 desaturase is the major desaturase present in the pheromone gland cells of H. assulta; whereas, the delta 11 desaturase is the major desaturase in pheromone glands of H. zea. It also appears that H. assulta lacks chain-shortening enzymes since stearic acid did not label any of the 16-carbon aldehydes. PMID- 12530204 TI - Adipokinetic hormones concentrations in the haemolymph of Schistocerca gregaria, measured by radioimmunoassay. AB - A new procedure for the measurement of adipokinetic hormone (AKH) concentrations in locust (Schistocerca gregaria) haemolymph is described: Haemolymph is extracted with chloroform/methanol/water and the aqueous layer is fractionated with reverse-phase cartridges and HPLC. The fractions corresponding to AKH-I (Lom AKH-I) and AKH-II (Scg-AKH-II) are then measured in a competitive binding assay using specific antibodies and [3H]AKHs. The procedures could be applied to any peptides containing N-terminal pyroglutamate residues including all members of the adipokinetic/hyperglycaemic/red pigment concentrating hormone family. Results show that the concentrations of both AKH-I and AKH-II increase within 5 min of initiation of flight and are maintained at approx. 15-fold (AKH-I) and 6-fold (AKH-II) the resting levels over flights of at least 60 min. Poisoning of locusts with either the insecticide deltamethrin or with potassium chloride also caused release of hormones. Starvation for 6 h caused elevation of hormone levels in 5th instar nymphs, but starvation for 6 or 20 h had little effect on hormones in adults, despite an increase in haemolymph diacylglycerols at 20 h. PMID- 12530205 TI - Site-directed mutagenesis and functional analysis of active site acidic amino acid residues D142, D144 and E146 in Manduca sexta (tobacco hornworm) chitinase. AB - Chitinases (EC 3.2.1.14) are glycosyl hydrolases that catalyze the hydrolysis of beta-(1, 4)-glycosidic bonds in chitin, the major structural polysaccharide present in the cuticle and gut peritrophic matrix of insects. Two conserved regions have been identified from amino acid sequence comparisons of family 18 glycosyl hydrolases, which includes Manduca sexta (tobacco hornworm) chitinase as a member. The second of these regions in M. sexta chitinase contains three very highly conserved acidic amino acid residues, D142, D144 and E146, that are probably active site residues. In this study the functional roles of these three residues were investigated using site-directed mutagenesis for their substitutions to other amino acids. Six mutant proteins, D142E, D142N, D144E, D144N, E146D and E146Q, as well as the wild-type enzyme, were produced using a baculovirus-insect cell line expression system. The proteins were purified by anion-exchange chromatography, after which their physical, kinetic and substrate binding properties were determined. Circular dichroism spectra of the mutant proteins were similar to that of the wild-type protein, indicating that the presence of mutations did not change the overall secondary structures. E146 was required for enzymatic activity because mutants E146Q and E146D were devoid of activity. D144E retained most of the enzymatic activity, but D144N lost nearly 90%. There was a shift in the pH optimum from alkaline pH to acidic pH for mutants D142N and D144E with minimal losses of activity relative to the wild-type enzyme. The pH-activity profile for the D142E mutation resembled that of the wild type enzyme except activity in the neutral and acidic range was lower. All of the mutant proteins bound to chitin. Therefore, none of these acidic residues was essential for substrate binding. The results indicate that E146 probably functions as an acid/base catalyst in the hydrolytic mechanism, as do homologous residues in other glycosyl hydrolases. D144 apparently functions as an electrostatic stabilizer of the positively charged transition state, whereas D142 probably influences the pKa values of D144 and E146. PMID- 12530206 TI - Molecular cloning and functional expression of a proline transporter from Manduca sexta. AB - We report the molecular cloning of a L-proline transporter, MasPROT cDNA and its splice variants MasPROT.16 and MasPROT.2 from the central nervous system of Manduca sexta. Sequence analysis revealed that MasPROT belongs to a family of high affinity Na+/Cl- dependent neurotransmitter transporters. The deduced amino acid (aa) sequence of 556 aa having an estimated molecular mass of 58.9 kDa is predicted to have 12 putative transmembrane domains (TMD) and a characteristic large extracellular loop between TMD3 and TMD4. Sequence comparison to other members of the family indicates that it falls into the glycine-proline transporter subfamily. Transiently expressed MasPROT cDNA in Xenopus oocytes exclusively transported proline. Northern analysis shows that it is expressed predominantly in central nervous system, however, low levels are present in midgut, hindgut and Malpighian tubules. Two mRNA transcripts of sizes 3.6 and 8 Kb were found in all tissues except hindgut, where only a smaller transcript exists. RT-PCR and Southern blot analysis revealed the presence of MasPROT transcripts in flight muscles but not in leg muscles. Our preliminary data suggests that this transporter is an insect homologue of mammalian proline transporters. MasPROT.16 is a short splice variant encoding for 174 amino acids and shares 138 amino acids from the N terminus of MasPROT. MasPROT.2 is a long splice variant that contains six introns that coincide precisely with the previously mapped exon/intron boundaries of the members of this superfamily. PMID- 12530207 TI - The kinase activity of the giant protein projectin of the flight muscle of Locusta migratoria. AB - Projectin is a member of the functionally and structurally heterogeneous family of myosin light chain kinases associated to myosin of synchronous as well as asynchronous insect muscles. We examined the phosphotransferase activity of projectin from flight muscle of Locusta migratoria. Isolated projectin exhibits an unstimulated autophosphorylation activity in vitro. We observed differences in the formation of synthetic filaments with myosin, and paramyosin depending on if projectin was autophosphorylated in vitro or not. Aggregates of native projectin with myosin and paramyosin (molar ratio 0.08:1:0.5) showed diameters 20-50 nm similar to those of myosin filaments. When in vitro autophosphorylated projectin was used we predominantly obtained, however, subfilament-like structures of only 7-10 nm in diameter. The in vitro autophosphorylation of projectin was suppressed in the presence of either acto-myosin, actin-filaments or myosin, but still seems to exhibit a phosphorylation activity: Projectin added to actomyosin resulted in the phosphorylation of three polypeptides of apparent molecular masses of 200, 165 and 100 kDa, respectively. These data suggest that the autophosphorylation activity of projectin is regulated by its environment. We conclude, therefore, a dual function of its kinase domain: at first, a role of its autophosphorylation in the formation of myosin filaments (association of subfilaments to filaments); secondly, the transphosphorylation activity of projectin modulates the contractile response of the actomyosin system by phosphorylating some of its components. Moreover, we could stimulate in vitro the projectin autophosphorylation 3.4-fold by calmodulin (EC50 = 17.8 nM). However, the transphosphorylations described above were not stimulated by calmodulin. PMID- 12530208 TI - Characterization of lipophorin binding to the fat body of Rhodnius prolixus. AB - In insects, lipids are transported by a hemolymphatic lipoprotein, lipophorin. The binding of lipophorin to the fat body of the hematophagous insect Rhodnius prolixus was characterized in a fat body membrane preparation, obtained from adult females. For the binding assay, purified lipophorin was radiolabelled in the protein moiety (125I-HDLp), and it was shown that iodination did not affect the affinity of the membrane preparation for lipophorin. Under incubation conditions used, lipophorin binding to membranes achieved equilibrium after 40-60 min, but this time was longer when a low concentration of lipophorin was present in the medium. The capacity of the fat body membrane preparation to bind lipophorin was abolished when membranes were pre-treated with trypsin, and it was also affected by heat. When 125I-HDLp was incubated with increasing concentrations of membrane protein, corresponding increases in binding were observed. Lipophorin binding was sensitive to pH, and it was maximal between pH 6.0 and 7.0. The specific binding of lipophorin to the fat body membrane preparation was a saturable process, with a Kd of 2.1 +/- 0.4 x 10(-7)M and a maximal binding capacity of 289 +/- 88 ng lipophorin/microgram of membrane protein. Binding to the fat body membranes did not depend on calcium, but it was affected by ionic strength, being totally inhibited at high salt concentrations. Suramin also interfered with lipophorin binding and it was abolished in the presence of 2 mM suramin, but at concentrations of 0.05 and 0.1 mM it seemed to increase binding activity slightly. Fat body membrane preparation from Rhodnius prolixus was able to bind lipophorin from Manduca sexta larvae. PMID- 12530209 TI - The major acid soluble proteins of adult female Anopheles darlingi salivary glands include a member of the D7-related family of proteins. AB - The salivary gland proteins of adult female Anopheles darlingi were fractionated by reverse-phase HPLC and the five major peaks were submitted for amino-terminal sequencing using automated Edman degradation. The amino acid sequence of one of the purified salivary gland proteins showed similarity with the D7r3 protein of An. gambiae. Cloning and sequencing of two cDNAs allowed the prediction of the complete sequence of the An. darlingi D7 protein. The D7r3 protein is present specifically in adult female salivary glands of An. darlingi and despite being one of the major salivary gland proteins its function is not known. Predictions of secondary and tertiary structures revealed the similarity of the An. darlingi D7 protein to insect odorant binding proteins. This suggests that D7 proteins may act as carriers of hydrophobic molecules in mosquito saliva. PMID- 12530210 TI - The major protein in the midgut of teneral Glossina morsitans morsitans is a molecular chaperone from the endosymbiotic bacterium Wigglesworthia glossinidia. AB - Molecules in the midgut of the tsetse fly (Diptera: Glossinidiae) are thought to play an important role in the life cycle of African trypanosomes by influencing their initial establishment in the midgut and subsequent differentiation events that ultimately affect parasite transmission. It is thus important to determine the molecular composition of the tsetse midgut to aid in understanding disease transmission by these medically important insect vectors. Here, we report that the most abundant protein in the midguts of teneral (unfed) Glossina morsitans morsitans is a 60 kDa molecular chaperone of bacterial origin. Two species of symbiotic bacteria reside in the tsetse midgut, Sodalis glossinidius and Wigglesworthia glossinidia. To determine the exact origin of the 60 kDa molecule, a protein microchemical approach involving two-dimensional (2-D) gel electrophoresis and mass spectrometry was used. Peptide mass maps were compared to virtual peptide maps predicted for S. glossinidius and W. glossinidia 60 kDa chaperone sequences. Four signature peptides were identified, revealing that the source of the chaperone was W. glossinidia. Comparative 2-D gel electrophoresis and immunoblotting further revealed that this protein was localized to the bacteriome and not the distal portion of the tsetse midgut. The possible function of this highly abundant endosymbiont chaperone in the tsetse midgut is discussed. PMID- 12530211 TI - Cattle tick Boophilus microplus salivary gland contains a thiol-activated metalloendopeptidase displaying kininase activity. AB - This work reports on the characterization of a metalloendopeptidase kininase present in Boophilus microplus salivary glands. Using the guinea pig ileum assay, salivary gland whole extracts (SGE) were found to have a potent kininase activity. Ion-exchange chromatography separated two kininase activities from SGE. The major enzymatic component, eluted at lower ionic strength, was named BooKase (Boophilus Kininase). Analysis of the hydrolysis products by capillary electrophoresis identified Phe5-Ser6 as the only hydrolyzable peptide bond in bradykinin after BooKase treatment. This is the same specificity as the mammalian thimet oligoendopeptidase (EC 3.4.24.15). Like this enzyme, BooKase is also a metallo-peptidase (requires Mn2+) and is activated by-SH protecting reagents. In addition, BooKase was partially inhibited by cFP-AAF-pAB, a specific inhibitor of thimet oligopeptidase. Contrary to other kininases, BooKase had no activity upon angiontensin I. Our results show that BooKase behaves as a typical peptidase with kinase activity. PMID- 12530212 TI - Chemical characterization of cytoplasmic lipid droplets in the pheromone producing cells of the silkmoth, Bombyx mori. AB - Accumulation of lipid droplets within the cytoplasm is a common feature of the pheromone gland cells of many lepidopteran species. The cytoplasmic lipid droplets in the pheromone-producing cells of the silkmoth, Bombyx mori, were effectively extracted by dipping the trimmed glands in acetone for 10 min. In order to analyze the components originating from the lipid droplets, we separated the acetone extracts prepared before and after adult eclosion using HPLC, and specified the peaks showing a similar pattern of stage-dependence to that in the morphological change of the lipid droplets previously reported by Fonagy et al. (Arthropod Struct. Dev. 30 (2001) 113). Finally, we specified the peaks #1-5 and #1a-4a separated by reversed-phase HPLC as lipid droplet contents. Structure elucidation using FAB-MS and MS-MS analyses confirmed that they were triacylglycerols (TGs), and 12 species of TGs were identified as lipid droplet contents. Fatty acyl groups contained in these TGs were limited to five unsaturated C16 and C18 fatty acyl groups (delta 11-hexadecenoate, delta 10,12 hexadecadienoate, delta 9-octadecenoate, delta 9,12-ocatadecadienoate, and delta 9,12,15-ocatadecatrienoate), including the pheromone precursor delta 10,12 hexadecadienoate as a major component. Digestion with porcine pancreatic lipase confirmed that three major TGs eluted in the peaks #3-5 all contained C18 fatty acyl groups at the sn-2 position, indicating that the pheromone precursor is sequestered preferentially at the sn-1 and/or sn-3 position. Present results combined with the fact that the morphological change of the lipid droplets is under the control of PBAN indicate that the role of the cytoplasmic lipid droplets in the pheromone-producing cells is to store the pheromone precursor in the form of TGs and to provide it for pheromone production in response to the external signal of PBAN. PMID- 12530213 TI - Molecular characterization of the insect immune protein hemolin and its high induction during embryonic diapause in the gypsy moth, Lymantria dispar. AB - During the embryonic (pharate first instar) diapause of the gypsy moth, Lymantria dispar, a 55 kDa protein is highly up-regulated in the gut. We now identify that protein as hemolin, an immune protein in the immunoglobulin superfamily. We isolated a gypsy moth hemolin cDNA and demonstrated a high degree of similarity with hemolins from three other moth species. Hemolin mRNA levels increased at the time of diapause initiation and remained high throughout the mandatory period of chilling required to terminate diapause in this species, and then dropped in late diapause. This mRNA pattern reflects the pattern of protein synthesis. These results suggest that hemolin is developmentally up-regulated in the gut during diapause. Diapause in this species can be prevented using KK-42, an imidazole derivative known to inhibit ecdysteroid biosynthesis, and gypsy moths treated in this manner failed to elevate hemolin mRNA. Conversely, this diapause appears to be initiated and maintained by the steroid hormone, 20-hydroxyecdysone, and the addition of 20-hydroxyecdysone to the culture medium elevated hemolin mRNA in the gut. Our results thus indicate a role for 20-hydroxyecdysone in the elevation of hemolin mRNA during diapause. Presumably, hemolin functions to protect the gypsy moth from microbial infection during its long, overwintering diapause. PMID- 12530214 TI - Vitellogenin of the cockroach, Leucophaea maderae: nucleotide sequence, structure and analysis of processing in the fat body and oocytes. AB - A cDNA encoding vitellogenin (Vg) of the cockroach, Leucophaea maderae was cloned and sequenced. The deduced amino acid sequence consisting of 1913 residues (including 15 residues for a putative signal peptide) was obtained. Amino terminal sequence analysis demonstrated that the pro-Vg was cleaved into four polypeptide 'subunits' following the three consensus RXXR cleavage site sequences, which were secreted as four Vg polypeptides (apparent molecular weights = 112-, 100-, 92- and 55-kD), sequestered, and deposited in the egg as four respective vitellin (Vn) polypeptides. There was, however, an additional 90 kD Vn polypeptide existed in the egg. We show that this polypeptide is a processed product from 92-kD Vn polypeptide. Northern blot analysis of poly (A)+ RNA reveals that mRNA coding for Vg is present only in the female fat body cells but neither in the ovary nor in the male fat body cells. The deduced amino acid sequence contained a serine-rich stretch at the C-terminal region. This stretch occurred also in Vgs of Periplaneta americana (Vg1 and Vg2) and Blattella germanica. The Vg of L. maderae had 26% and 31% homology with those of P. americana (Vg1 and Vg2) and B. germanica, respectively. Phylogenetic analysis (neighbour-joining) was made using four cockroach Vgs and the tree was compared with other molecular and conventional phylogenetic trees. PMID- 12530215 TI - Site-directed mutagenesis and functional analysis of an active site tryptophan of insect chitinase. AB - Chitinase is an enzyme used by insects to degrade the structural polysaccharide, chitin, during the molting process. Tryptophan 145 (W145) of Manduca sexta (tobacco hornworm) chitinase is a highly conserved residue found within a second conserved region of family 18 chitinases. It is located between aspartate 144 (D144) and glutamate 146 (E146), which are putative catalytic residues. The role of the active site residue, W145, in M. sexta chitinase catalysis was investigated by site-directed mutagenesis. W145 was mutated to phenylalanine (F), tyrosine (Y), isoleucine (I), histidine (H), and glycine (G). Wild-type and mutant forms of M. sexta chitinases were expressed in a baculovirus-insect cell line system. The chitinases secreted into the medium were purified and characterized by analyzing their catalytic activity and substrate or inhibitor binding properties. The wild-type chitinase was most active in the alkaline pH range. Several of the mutations resulted in a narrowing of the range of pH over which the enzyme hydrolyzed the polymeric substrate, CM-Chitin-RBV, predominantly on the alkaline side of the pH optimum curve. The range was reduced by about 1 pH unit for W145I and W145Y and by about 2 units for W145H and W145F. The W145G mutation was inactive. Therefore, the hydrophobicity of W145 appears to be critical for maintaining an abnormal pKa of a catalytic residue, which extends the activity further into the alkaline range. All of the mutant enzymes bound to chitin, suggesting that W145 was not essential for binding to chitin. However, the small difference in Km's of mutated enzymes compared to Km values of the wild type chitinase towards both the oligomeric and polymeric substrates suggested that W145 is not essential for substrate binding but probably influences the ionization of a catalytically important group(s). The variations in kcat's among the mutated enzymes and the IC50 for the transition state inhibitor analog, allosamidin, indicate that W145 also influences formation of the transition state during catalysis. PMID- 12530218 TI - Expression and characterization of a novel teratocyte protein of the braconid, Microplitis croceipes (cresson). AB - Microplitis croceipes wasps overcome host immunity by inducing changes in host physiology using factors derived from the embryo and/or larva. Teratocytes of some parasitic wasps circulate in the host hemolymph after egg hatch and synthesize proteins (TSPs), some of which are secreted to alter host physiology in support of endoparasitoid development. TSPs appear to alter host physiology, at least in part, by inhibiting synthesis of certain proteins. M. croceipes teratocytes synthesize a 13.9 kDa protein (TSP14), which inhibits synthesis of host proteins that are linked to larval growth and development. A cDNA encoding TSP14 was generated by RT-PCR from teratocyte RNA, and cloned into yeast expression vectors to produce sufficient recombinant protein for functional analyses. RecTSP14 was produced using the yeast expression system at a concentration of more than 300 micrograms/L. The recTSP14 inhibited in vitro translation of larval Heliothis virescens RNA, with the activity sensitive to boiling, protein concentration, incubation time, and storage temperatures. Although recTSP14 inhibited translation of some cellular RNAs in vitro, the in vivo incorporation of [35S]-methionine into proteins of selected insect and mammalian cell lines was not inhibited. These findings suggest that recTSP14 entry is cell type-specific and required to inhibit synthesis of target protein(s). PMID- 12530216 TI - Moth desaturase characterized that produces both Z and E isomers of delta 11 tetradecenoic acids. AB - The redbanded leafroller moth, Argyrotaenia velutinana (Lepidoptera: Tortricidae) uses a 92:8 mixture of (Z)-11- and (E)-11-tetradecenyl acetate in its pheromone blend. These are produced in the abdominal pheromone gland from the corresponding acids, which are biosynthesized in the gland in a 3:2 Z/E ratio by desaturation of myristoyl CoA. The delta 11 desaturase involved in this reaction exhibits unusual substrate and stereospecificities in specifically producing Z11 and E11 isomers of tetradecenoic acid, and exhibiting no activity with C16 and C18 precursor acids. This report describes the cloning and expression of the redbanded leafroller moth delta 11 desaturase, and compares its amino-acid sequence to those of other known insect Z9, Z10, Z11, and E11 desaturases. The metabolic Z9 desaturase from fat body tissue also was cloned and expressed, and found mainly to produce Z9-16:Acid and Z9-18:Acid. The open reading frame of the delta 11 desaturase encodes a protein with 329 amino acids, whereas the open reading frame of the Z9 desaturase encodes a protein with 351 amino acids. Addition of this new delta 11 desaturase with its different substrate and regiospecificites to the databank of characterized integral-membrane desaturases will be key in efforts to determine amino-acid mutations responsible for the wide array of unsaturated fatty-acid products. PMID- 12530217 TI - Sequence of a cDNA and expression of the gene encoding a putative epidermal chitin synthase of Manduca sexta. AB - Glycosyltransferases are enzymes that synthesize oligosaccharides, polysaccharides and glycoconjugates. One type of glycosyltransferase is chitin synthase, a very important enzyme in biology, which is utilized by insects, fungi, and other invertebrates to produce chitin, a polysaccharide of beta-1,4 linked N-acetylglucosamine. Chitin is an important component of the insect's exoskeletal cuticle and gut lining. To identify and characterize a chitin synthase gene of the tobacco hornworm, Manduca sexta, degenerate primers were designed from two highly conserved regions in fungal and nematode chitin synthase protein sequences and then used to amplify a similar region from Manduca cDNA. A full-length cDNA of 5152 nucleotides was assembled for the putative Manduca chitin synthase gene, MsCHS1, and sequencing of genomic DNA verified the contiguity of the sequence. The MsCHS1 cDNA has an ORF of 4692 nucleotides that encodes a transmembrane protein of 1564 amino acid residues with a mass of approximately 179 kDa (GenBank no. AY062175). It is most similar, over its entire length of protein sequence, to putative chitin synthases from other insects and nematodes, with 68% identity to enzymes from both the blow fly, Lucilia cuprina, and the fruit fly, Drosophila melanogaster. The similarity with fungal chitin synthases is restricted to the putative catalytic domain, and the MsCHS1 protein has, at equivalent positions, several amino acids that are essential for activity as revealed by mutagenesis of the fungal enzymes. A 5.3-kb transcript of MsCHS1 was identified by northern blot hybridization of RNA from larval epidermis, suggesting that the enzyme functions to make chitin deposited in the cuticle. Further examination by RT-PCR showed that MsCHS1 expression is regulated in the epidermis, with the amount of transcript increasing during phases of cuticle deposition. PMID- 12530219 TI - Characterization of an extracellular salicyl alcohol oxidase from larval defensive secretions of Chrysomela populi and Phratora vitellinae (Chrysomelina). AB - Larvae of a number of chrysomelid leaf beetles sequester phenol glucosides such as salicin from their food plants, i.e. Salix and Populus spp. Salicin is hydrolyzed in the glandular reservoir of the defensive glands. The resulting salicyl alcohol (saligenin) is oxidized by an extracellular oxidase. The product salicylaldehyde accumulates as major defensive compound. The secretions from Chrysomela populi and Phratora vitellinae were preserved in saturated ammonium sulfate solution and subjected to micro-purification of the oxidase by means of electrophoretic methods. The enzyme from P. vitellinae has a native M(r) of 334,000 and a subunit M(r) of 79,000 indicating a tetrameric enzyme. The isoelectric points of the enzymes from C. populi and P. vitellinae are at pH 5.4 and 5.2, respectively. In the oxidation of salicyl alcohol oxygen functions as electron acceptor yielding hydrogen peroxide as product. Hydrogen peroxide does not accumulate in native secretions but appears to be degraded most likely by a catalase. The oxidases from the two species show broad pH optima in the range 5.5 to 6.5, they oxidize salicyl alcohol as main substrate. Minor substrates are several ortho-substituted and to a lesser extent meta- but not para-substituted benzyl alcohols. In the presence of 8-hydroxygeraniol only trace amounts of the respective aldehyde are formed. The Km values of salicyl alcohol are 132 mM (C. populi) and 63 mM (P. vitellinae). The extracellular enzyme, which is functionally related to fungal aryl alcohol oxidase (EC 1.1.3.7) and vanillyl alcohol oxidase (EC 1.1.3.38) was named salicyl alcohol oxidase. The continuous formation of salicylaldehyde in the glandular reservoir can be compared to the operation of an enzyme reactor. Due to its low aqueous solubility the produced aldehyde steadily leaves the aqueous reaction fluid and builds up an organic phase which may account for 15% of the total liquid volume of the secretion. PMID- 12530220 TI - Male jeffrey pine beetle, Dendroctonus jeffreyi, synthesizes the pheromone component frontalin in anterior midgut tissue. AB - The male Jeffrey pine beetle, Dendroctonus jeffreyi Hopkins (Coleoptera: Scolytidae), produces the bicyclic ketal frontalin as part of a complex semiochemical blend. A key regulated enzyme in the mevalonate pathway, 3-hydroxy 3-methylglutaryl-CoA reductase (HMG-R), showed high transcript levels in the anterior midgut of male Jeffrey pine beetles by in situ hybridization. HMG-R expression in this area of the alimentary canal was related to male emergence, where emerged males demonstrated significant up-regulation of HMG-R transcript and pre-emerged males showed only basal levels. Pre-emerged males were induced to express high levels of HMG-R transcript by treatment with juvenile hormone (JH) III. Additionally, isolated anterior midgut tissue from JH III-treated males converted radiolabeled acetate to frontalin, as assayed by radio-HPLC, providing strong evidence that this is the site of frontalin production in male beetles. PMID- 12530221 TI - On the biosynthesis of Rhodnius prolixus heme-binding protein. AB - The biosynthesis of Rhodnius prolixus heme-binding protein (RHBP), which is present in the hemolymph and oocytes of Rhodnius prolixus, was investigated. Fat bodies of female insects incubated in vitro with 14C-leucine were able to synthesize and secrete 14C-RHBP to the culture medium. Titrtion of synthesized RHBP with hemin showed that the protein secreted by the fat bodies is bound to heme, despite the presence of apo-RHBP in the hemolymph. The sequence of the RHBP cDNA encodes a pre-protein of 128 amino acids with no significant homology to any known protein. Northern-blot assays revealed that RHBP expression was limited to fat bodies. The levels of both RHBP mRNA and secreted protein increased in response to blood meal. In addition, the time-course of RHBP secretion in vitro paralleled mRNA accumulation observed in vivo. The inhibition of the de novo heme biosynthesis by treatment of fat bodies with succinyl acetone (SA), an irreversible inhibitor of delta-aminolevulinic acid-dehydratase, led to a significant decrease of heme-RHBP secretion. Nevertheless, the levels of RHBP mRNA were not modified by SA treatment, suggesting that the heme availability is involved in a post-transcriptional control of the RHBP synthesis. PMID- 12530222 TI - Pheromone binding proteins of Epiphyas postvittana (Lepidoptera: Tortricidae) are encoded at a single locus. AB - The light brown apple moth, Epiphyas postvittana (Tortricidae: Lepidoptera) uses a blend of (E)-11-tetradecenyl acetate and (E,E)-9,11-tetradecadienyl acetate as its sex pheromone. Odorant binding proteins, abundant in the antennae of male and female E. postvittana, were separated by native PAGE to reveal four major proteins with distinct mobilities. Microsequencing of their N-terminal residues showed that two were general odorant binding proteins (GOBPs) while two were pheromone binding proteins (PBPs). Full length cDNAs encoding these proteins were amplified using a combination of PCR and RACE-PCR. Sequence of the GOBPs revealed two genes (EposGOBP1, EposGOBP2), similar to orthologues in other species of Lepidoptera. Eleven cDNAs of the PBP gene were amplified, cloned and sequenced revealing two major phylogenetic clusters of PBP sequences differing by six amino acid substitutions. The position of the six amino acid differences on the protein was predicted by mapping onto the three-dimensional structure of PBP of Bombyx mori. All six substitutions were predicted to fall on the outside of the protein away from the inner pheromone binding pocket. One substitution does fall close to the putative dimerisation region of the protein (Ser63Thr). Expression of three of the cDNAs in a baculovirus expression system revealed that one class encodes an electrophoretically slow form (EposPBP1-12) while the other encodes a fast form (EposPBP1-2, EposPBP1-3). A native Western of these expressed proteins compared with antennal protein extracts demonstrated that PBP is also expressed in female antennae and that PBP may be present as a dimer as well as a monomer in E. postvittana. The fast and slow forms of EposPBP1 are allelic. Westerns on single antennal pair protein extracts and allele-specific PCR from genomic DNA both show a segregating pattern of inheritance in laboratory and wild populations. Radio labelled (E)-11-tetradecenyl acetate binds to both fast and slow PBP forms in gel assays. Taken together, the genetic and biochemical data do not support the hypothesis that these PBPs are specific for each component of the E. postvittana pheromone. However, duplication of this PBP locus in the future might allow such diversification to evolve, as observed in the other species. PMID- 12530224 TI - Sequences of elongation factors-1 alpha and -1 gamma and stimulation by juvenile hormone in Locusta migratoria. AB - Two cDNAs encoding the alpha and gamma subunits of translation elongation factor 1 (EF-1) have been cloned and sequenced from the African migratory locust, Locusta migratoria. Southern blotting and real-time PCR analyses indicated that these sequences represent single copy genes. Comparison with sequences from other species indicated greater conservation for EF-1 alpha than for EF-1 gamma. The developmental profiles for EF-1 alpha and -1 gamma mRNA expression in the fat body paralleled reported changes in the hemolymph juvenile hormone (JH) titer in the fifth instar and were elevated during early reproductive maturation in the female adult. In maturing adults, there was a greater accumulation of EF-1 alpha and -1 gamma transcripts in females than in males. The levels of both transcripts were greatly increased by an enriched diet, previously shown to elevate JH titers and accelerate vitellogenin production. Treating JH-deprived adult females with the JH analog, methoprene, resulted in more than doubling of transcript levels of both genes, supporting the hypothesis that JH could stimulate the accumulation of LmEF-1 alpha and -1 gamma transcripts. We suggest that production of elongation factors, increased by JH, may contribute to the massive protein synthesis required for egg production. PMID- 12530223 TI - Juvenile hormone signaling during oogenesis in Drosophila melanogaster. AB - Juvenile hormone (JH) participates both in the control of insect development and the establishment of reproductive maturity. In cultured Drosophila cells and in ovarian nurse cells, JH and its synthetic analog, methoprene, induce the expression of two related genes. These genes encode highly similar amino acid transport proteins that are homologous to transporters found in a variety of eukaryotes. JhI-21 is a novel Drosophila gene, and minidiscs (mnd) is a gene that was identified earlier. Two JH-inducible genes are regulated by different molecular mechanisms; JhI-21 behaves as a secondary JH-responsive gene, while mnd behaves as a primary responsive gene. Both JhI-21 and mnd transcripts show developmental profiles, which are consistent with JH regulation. Following eclosion, transcripts from JhI-21 and mnd are synthesized in ovarian nurse cells and subsequently sequestered in the mature egg. Their ectopic accumulation in ovaries can be induced by topical methoprene application. In apterous (ap4) mutant adults defective in JH secretion, mnd and JhI-21 RNA levels are severely reduced, but normal abundance is rescued to a high degree by topical methoprene treatment. Based on the evidence, we propose that during sexual maturation of Drosophila, JH provides a signal to the ovary that leads to the production of several maternally inherited mRNAs. PMID- 12530225 TI - A structural model of the chitin-binding domain of cuticle proteins. AB - The nature of the interaction of insect cuticular proteins and chitin is unknown even though about half of the cuticular proteins sequenced thus far share a consensus region that has been predicted to be the site of chitin binding. We previously predicted the preponderance of beta-pleated sheet in the consensus region and proposed its responsibility for the formation of helicoidal cuticle (Iconomidou et al., Insect Biochem. Mol. Biol. 29 (1999) 285). Consequently, we have also verified experimentally the abundance of antiparallel beta-pleated sheet in the structure of cuticle proteins (Iconomidou et al., Insect Biochem. Mol. Biol. 31 (2001) 877). In this work, based on sequence and secondary structure similarity of cuticle proteins, and especially that of the consensus motif, to that of bovine plasma retinol binding protein (RBP), we propose by homology modelling an antiparallel beta-sheet half-barrel structure as the basic folding motif of cuticle proteins. This folding motif may provide the template for elucidating cuticle protein-chitin interactions in detail and reveal the precise geometrical formation of cuticle's helicoidal architecture. This predicted motif is another example where nature utilizes an almost flat protein surface covered by aromatic side chains to interact with the polysaccharide chains of chitin. PMID- 12530226 TI - Changes in cysteine protease activity and localization during midgut metamorphosis in the crucifer root maggot (Delia radicum). AB - We show that differential localization and/or activation of two cysteine protease activities occur at the onset of dipteran midgut metamorphosis. A 26 kDa cysteine protease activity was associated specifically with midgut tissues of late third instar larvae. Starvation of mid third instar larvae simulated the onset of prepupation and resulted in loss of the 26 kDa protease activity. A cDNA clone encoding a cysteine protease, termed DrCP1, was isolated and shown to be highly similar to those from Sarcophaga peregrina and Drosophila melanogaster (DmCP1). DrCP1 mRNA was present in all developmental stages including eggs, larvae, pupae and adults, but was highly induced at the onset of the larval-pupal transition and thereafter. The DrCP1 protein is localized to the exterior of the midgut tissues during the onset of the prepupal transition period, possibly in response to ecdysone. Analysis of transcription factor binding sites associated with the DmCP1 promoter indicated that elements exist that allow for both ecdysone mediated as well as tissue-specific regulation. Based upon these and other studies we propose: (1) that the expression, activity and localization of the DrCP1-like cysteine proteases are highly regulated throughout development; and, (2) that cysteine protease activities are involved in aspects of tissue reconstruction at the onset of and during metamorphosis. PMID- 12530227 TI - Imaging of the upper cranial nerves I, III-VIII, and the cavernous sinuses. AB - This article discusses the upper cranial nerves (I, III-VIII) and their anatomy as it pertains to intra-axial nuclei and tracts, cisternal portions, and extracranial portions. In addition, the most common pathologic processes affecting the upper cranial nerves are discussed and illustrated. Because the evaluation of small structures requires imaging techniques that provide high resolution and contrast, MR imaging is the examination of choice. CT still plays a limited but important role in the evaluation of intraosseous portions of some cranial nerves. PMID- 12530228 TI - Imaging of the lower cranial nerves. AB - Knowledge of the nuclear origins and the normal course of the cranial nerves, and their functional components, is important in any evaluation of cranial neuropathy. MR imaging allows detailed evaluation of cranial nerve anatomy and pathology. Newer MR sequences allow more sensitive methods of detecting pathology and determining the cause of cranial neuropathy. The correlation of clinical findings with MR images will allow for more definitive evaluation. PMID- 12530229 TI - Laryngeal imaging. AB - Knowing the surgical options for treating laryngeal carcinomas and the factors that are used to select patients for radiation therapy leads to a more comprehensive interpretation of neck scans in patients with laryngeal tumors (Table 6). Critical factors include tumor volume; cartilaginous invasion; spread across supraglottic-glottic-subglottic [table: see text] boundaries; infiltration of preepiglottic, paraglottic, and pharyngeal planes; and nodal disease. MR imaging offers greater sensitivity to cartilaginous invasion than CT but leads to a high rate of false-positive studies, which decreases its overall accuracy. Thin section CT with multiplanar capability is competitive with direct coronal MR scanning and benefits from high specificity and submillimeter section thickness, if multidetector units are employed. Overall, the head and neck radiologist plays an invaluable role in assessing the extent of disease and therefore influences the appropriate selection from the available treatment options. PMID- 12530230 TI - Imaging of neoplasms of the paranasal sinuses. AB - The assessment of sinonasal malignancies requires a multidisciplinary team approach. Advances in pretherapeutic imaging have significantly contributed to the management of sinonasal tumors. CT and MR imaging play complementary roles in the assessment and staging of these malignancies by determining the presence or absence of extension of disease into the skull base and its foramina, the orbit, and the intracranial compartment. PMID- 12530231 TI - Imaging of the hypopharynx and cervical esophagus. AB - Cross-sectional imaging with CT and MR imaging plays a crucial role in the evaluation of cancers of the hypopharynx and cervical esophagus. Clinical examination, including endoscopy, often fails to detect the full extent of this tumor as it has high propensity to grow in submucosal fashion. Detailed description of the cross-sectional anatomy and of the tumor growth pattern will aid in the detection and evaluation of the full extent of the lesion. Post treatment imaging findings are addressed. PMID- 12530232 TI - MR imaging of perineural tumor spread. AB - Perineural tumor spread (PNS) is a common and potentially devastating complication of head and neck cancer. Because perineural tumor spread may be asymptomatic, the radiologist plays a critical role in the evaluation of the head and neck cancer patient. Although PNS may be seen on CT, MR imaging is far more sensitive and is the imaging modality of choice in the evaluation of perineural tumor spread. PMID- 12530233 TI - MR imaging of lymph nodes in the head and neck. AB - This article reviews imaging-based nodal classification, recent advancements in MR techniques for lymph node imaging, and characteristics of MR imaging for neck lymphadenopathy. Further technical development of MR imaging is necessary to improve accuracy of lymph node metastases in patients with N0 neck. Development of new tissue-specific MR contrast agents and functional imaging studies to address biologic activity of lymph nodes are essential in the future. PMID- 12530234 TI - Life before RNA. AB - The hypothesis that life originated and evolved from linear informational molecules capable of facilitating their own catalytic replication is deeply entrenched. However, widespread acceptance of this paradigm seems oblivious to a lack of direct experimental support. Here, we outline the fundamental objections to the de novo appearance of linear, self-replicating polymers and examine an alternative hypothesis of template-directed coding of peptide catalysts by adsorbed purine bases. The bases (which encode biological information in modern nucleic acids) spontaneously self-organize into two-dimensional molecular solids adsorbed to the uncharged surfaces of crystalline minerals; their molecular arrangement is specified by hydrogen bonding rules between adjacent molecules and can possess the aperiodic complexity to encode putative protobiological information. The persistence of such information through self-reproduction, together with the capacity of adsorbed bases to exhibit enantiomorphism and effect amino acid discrimination, would seem to provide the necessary machinery for a primitive genetic coding mechanism. PMID- 12530235 TI - Archaeal genome organization and stress responses: implications for the origin and evolution of cellular life. AB - For DNA to be used as an informational molecule it must exist in the cell on the edge of stability because all genomic processes require local controlled melting. This presents mechanistic opportunities and problems for genomic DNA from hyperthermophilic organisms, whose unpackaged DNA could melt at optimal temperatures for growth. Hyperthermophiles are suggested to employ the novel positively supercoiling topoisomerase enzyme reverse gyrase (RG) to form positively supercoiled DNA that is intrinsically resistant to thermal denaturation. RG is presently the only archaeal gene that is uniquely found in hyperthermophiles and therefore is central to hypotheses suggesting a hypothermophilic origin of life. However, the suggestion that RG has evolved by the fusion of two pre-existing enzymes has led to hypotheses for a lower temperature for the origin of life. In addition to the action of topoisomerases, DNA packaging and the intracellular ionic environment can also manipulate DNA topology significantly. In the Euryarchaeota, nucleosomes containing minimal histones can adopt two alternate DNA topologies in a salt-dependent manner. From this we hypothesize that since internal salt concentrations are increased following an increase in temperature, the genomic effects of temperature fluctuations could also be accommodated by changes in nucleosome organization. In addition, stress-induced changes in the nucleoid proteins could also play a role in maintaining the genome in the optimal topological state in changing environments. The function of these systems could therefore be central to temperature adaptation and thus be implicated in origin of life scenarios involving hyperthermophiles. PMID- 12530236 TI - A culture-independent survey of the bacterial community in a radon hot spring. AB - Paralana is an active, radon-containing hot spring situated in a region of South Australia's Flinders Ranges with a long history of hydrothermal activity. Our aim was to determine the bacterial composition of Paralana using a culture independent, 16S rRNA-based technique. The presence of a diverse bacterial community was strongly suggested by the large number (approximately 180) of different ribotypes obtained upon analysis of nine hot spring samples. DNA sequencing of Paralana 16S rRNA genes corroborated this observation, identifying representatives of seven confirmed and two candidate divisions of the domain Bacteria. These included Cyanobacteria, Proteobacteria (both beta and delta subdivisions), the Cytophaga-Flexibacter-Bacteroides group, Low G + C Gram positives, Nitrospira, green non-sulfur bacteria, green sulfur bacteria, OP8, and OP12. No known ionizing radiation-resistant Bacteria were identified. Only one Paralana 16S rRNA sequence type (recombinant B5D) was homologous to a sequence previously identified from a radioactive environment. PMID- 12530237 TI - Molecular identification of cyanobacteria associated with stromatolites from distinct geographical locations. AB - Modern stromatolites represent a significant resource for studying microbial ecology and evolution. A preliminary investigation was undertaken employing specific genetic probes to characterize the cyanobacteria responsible for stromatolite construction in a range of environments, including microbial mats found in Australia not previously examined with molecular methods. Isolates of cyanobacteria were collected from stromatolites in thermal springs, hypersaline lakes, and oceanic fringes on two continents. A polymerase chain reaction specific for DNA of cyanobacterial 16S rRNA was developed, the resulting products of the DNA amplification reaction were sequenced, and the data were used to infer relatedness between the isolates studied and other members of the cyanobacterial radiation. Complete sequence was generated for the region from position 27 to 408 for 13 strains of cyanobacteria associated with stromatolites. All stromatolite derived sequences were most closely related to cyanobacteria, as indicated by local sequence alignment. It was possible to correlate genetic identity with morphological nomenclatures and to expand the phylogeny of benthic cyanobacteria. These inferences were also expanded to temporal variation in the dominant resident cyanobacterial species based on sampling of surface and core sinter laminations. Under the methods employed, only one cyanobacterial strain was detected in each sample, suggesting the possible dominance of a specific clonal population of cyanobacteria at any one time in the biota of the samples tested. The data indicate that internal core samples of a stromatolite at least 10 years old can be successfully analyzed by DNA-based methods to identify preserved cyanobacteria. PMID- 12530238 TI - Extremophiles and the search for extraterrestrial life. AB - Extremophiles thrive in ice, boiling water, acid, the water core of nuclear reactors, salt crystals, and toxic waste and in a range of other extreme habitats that were previously thought to be inhospitable for life. Extremophiles include representatives of all three domains (Bacteria, Archaea, and Eucarya); however, the majority are microorganisms, and a high proportion of these are Archaea. Knowledge of extremophile habitats is expanding the number and types of extraterrestrial locations that may be targeted for exploration. In addition, contemporary biological studies are being fueled by the increasing availability of genome sequences and associated functional studies of extremophiles. This is leading to the identification of new biomarkers, an accurate assessment of cellular evolution, insight into the ability of microorganisms to survive in meteorites and during periods of global extinction, and knowledge of how to process and examine environmental samples to detect viable life forms. This paper evaluates extremophiles and extreme environments in the context of astrobiology and the search for extraterrestrial life. PMID- 12530239 TI - Does the rapid appearance of life on Earth suggest that life is common in the universe? AB - It is sometimes assumed that the rapidity of biogenesis on Earth suggests that life is common in the Universe. Here we critically examine the assumptions inherent in this if-life-evolved-rapidly-life-must-be-common argument. We use the observational constraints on the rapidity of biogenesis on Earth to infer the probability of biogenesis on terrestrial planets with the same unknown probability of biogenesis as the Earth. We find that on such planets, older than approximately 1 Gyr, the probability of biogenesis is > 13% at the 95% confidence level. This quantifies an important term in the Drake Equation but does not necessarily mean that life is common in the Universe. PMID- 12530240 TI - A search for optical beacons: implications of null results. AB - Over the past few years a series of searches for interstellar radio beacons have taken place using the Parkes radio telescope. Here we report hitherto unpublished results from a search for optical beacons from 60 solar-type stars using the Perth-Lowell telescope. We discuss the significance of the null results from these searches, all of which were based on the interstellar contact channel hypothesis. While the null results of all searches to date can be explained simply by the nonexistence of electromagnetically communicating life elsewhere in the Milky Way, four other possible explanations that do not preclude its existence are proposed: (1) Extraterrestrial civilizations desiring to make contact through the use of electromagnetic beacons have a very low density in the Milky Way. (2) The interstellar contact channel hypothesis is incorrect, and beacons exist at frequencies that have not yet been searched. (3) The search has been incomplete in terms of sensitivity and/or target directions: Beacons exist, but more sensitive equipment and/or more searching is needed to achieve success. (4) The search has occurred before beacon signals can be expected to have arrived at the Earth, and beacon signals may be expected in the future. Based on consideration of the technology required for extraterrestrial civilizations to identify target planets, we argue that the fourth possibility is likely to be valid and that powerful, easily detectable beacons could be received in coming centuries. PMID- 12530241 TI - Active polar gullies on Mars and the role of carbon dioxide. AB - The detection of geologically recent channels and gullies on Mars has been interpreted as evidence for recent water activity on this arid and cold planet. The presence of active water would have considerable implications for the history of Mars and for the potential for an active near-surface biota; however, water is not the only substance that can flow and may actually be one of the least likely candidates when the modern permafrost environment is considered. Here, a key site of recent gully and channel development on Mars is reported at 71 degrees S in Sisyphi Cavi, within the annual zone of polar CO2 ice and snow accumulation. Superposition of channel features over and/or through the defrosting CO2 snowpack shows that the channels are active at the present day and probably have fluid flows every spring during the annual defrosting. In itself, this is a significant observation as active fluid flows of any nature have not yet been proven on Mars. However, the ambient temperature at the time of gully activity appears to require a role for CO2 in the formation of the channels, rather than water. A model is proposed for gas-lubricated flow in the channels based on avalanching of CO2 snowpack and clastic debris. If similar mechanisms are responsible for all the recent gullies on Mars, then perhaps no near-surface astrobiological targets are available. PMID- 12530242 TI - The observational case for Jupiter being a typical massive planet. AB - We identify a subsample of the recently detected extrasolar planets that is minimally affected by the selection effects of the Doppler detection method. With a simple analysis we quantify trends in the surface density of this subsample in the period-Msin(i) plane. A modest extrapolation of these trends puts Jupiter in the most densely occupied region of this parameter space, thus indicating that Jupiter is a typical massive planet rather than an outlier. Our analysis suggests that Jupiter is more typical than indicated by previous analyses. For example, instead of MJup mass exoplanets being twice as common as 2 MJup exoplanets, we find they are three times as common. PMID- 12530243 TI - Application of hyperspectral infrared analysis of hydrothermal alteration on Earth and Mars. AB - An integrated analysis of both airborne and field short-wave infrared hyperspectral measurements was used in conjunction with conventional field mapping techniques to map hydrothermal alteration in the central portion of the Mount Painter Inlier in the Flinders Ranges, South Australia. The airborne hyperspectral data show the spatial distribution of spectrally distinct minerals occurring as primary minerals and as weathering and alteration products. Field spectral measurements, taken with a portable infrared mineral analyzer spectrometer and supported by thin-section analyses, were used to verify the mineral maps and enhance the level of information obtainable from the airborne data. Hydrothermal alteration zones were identified and mapped separately from the background weathering signals. A main zone of alteration, coinciding with the Paralana Fault zone, was recognized, and found to contain kaolinite, muscovite, biotite, and K-feldspar. A small spectral variation associated with a ring-like feature around Mount Painter was tentatively determined to be halloysite and interpreted to represent a separate hydrothermal fluid and fluid source, and probably a separate system. The older parts of the alteration system are tentatively dated as Permo-Carboniferous. The remote sensing of alteration at Mount Painter confirms that hyperspectral imaging techniques can produce accurate mineralogical maps with significant details that can be used to identify and map hydrothermal activity. Application of hyperspectral surveys such as that conducted at Mount Painter would be likely to provide similar detail about putative hydrothermal deposits on Mars. PMID- 12530244 TI - Morphology: an ambiguous indicator of biogenicity. AB - This paper deals with the difficulty of decoding the origins of natural structures through the study of their morphological features. We focus on the case of primitive life detection, where it is clear that the principles of comparative anatomy cannot be applied. A range of inorganic processes are described that result in morphologies emulating biological shapes, with particular emphasis on geochemically plausible processes. In particular, the formation of inorganic biomorphs in alkaline silica-rich environments are described in detail. PMID- 12530245 TI - A clinical case for quality. Rhode Island takes the lead in first wave of initiatives seeking consensus on clinical practices as markers for quality care. AB - Clinical guidelines for treating three common conditions that send people to the hospital are becoming the initial focus of a wave of efforts to reach a consensus on clinical practices as markers for quality. Marcia Petrillo (left) leads Rhode Island's quality-improvement organization. Shown on the cover are Patricia Nolan, seated, state health director, standing from left, Petrillo; Cathy Boni of the state hospital association; and Maureen Maigret of the lieutenant governor's office. PMID- 12530246 TI - AMA paid CEO $1 million for last six months on job. PMID- 12530247 TI - An inside look. RSNA attendee marvels at new technologies. PMID- 12530248 TI - On the lookout. CMS studies outlier billing practices. PMID- 12530249 TI - The case of the vanishing giveback. Unraveling the mystery of how docs managed to leave the Capitol empty-handed. PMID- 12530250 TI - Leaders of the pack. Solucient's annual list of the nation's 100 top hospitals features organizations that take excellence several steps further. PMID- 12530251 TI - Is a baby clone on board? PMID- 12530252 TI - Abducting the cloning debate. PMID- 12530253 TI - Sick, tired and not taking it anymore. PMID- 12530254 TI - Never say diet. PMID- 12530255 TI - [Genetic translocations in oncohematology]. PMID- 12530256 TI - [Genetic disorders in cutaneous melanoma]. PMID- 12530257 TI - [Role of phosphatidylinositol-3-kinase in the development of cancer]. PMID- 12530258 TI - [Computerized and magnetic resonance tomography in the staging of lung cancer]. AB - CT (157) and MRI (74) potential in lung cancer staging was studied. Central form was identified in 195 patients (75%). The data were compared in 34. Comparison with surgical evidence was carried out in 47 (CT) and 35 (MRI). Both procedures proved valuable. Use of CT cut down the gap between the results of clinical and surgical evaluation to 17%. Higher precision was achieved by MRI application which was demonstrated by a comparison of the sensitivities of both procedures attained in evaluating the three parameters. The sensitivities of MRI (96%) and CT (90%) in the detection of intrathoracic lymphadenopathy were almost identical. MRI was superior to CT in detecting large vessel involvements (sensitivities--91 and 42%, respectively) as well as those of the pleura, pericardium and thoracic wall. PMID- 12530259 TI - [Triiodothyronine and thyroxin binding to red blood cells in children and adolescents with thyroid cancer]. AB - A comparison of triiodothyronine and thyroxin binding to red blood cells sampled from children and adolescents with such thyroid pathologies as cancer, adenoma, nodular goiter and thyroiditis, on the one hand, and healthy children, on the other, has established significant changes in the hormone-binding activity of erythrocytes in thyroid cancer patients. The thyroiditis-related changes suggested that disturbed thyroid hormone-binding by blood cells might play a role in thyroiditis and thyroid cancer pathogenesis. PMID- 12530260 TI - [Clinical and cytogenetic investigation of a family with high predisposition for intestinal polyps]. AB - A medico-genetic investigation of a family, consisting of 25 members, revealed high predisposition to malignant pathology of the gastrointestinal tract: rectal cancer was diagnosed in 2, malignant polyposis of the large intestine--1, diffuse polyposis of the large intestine (DPLI)--3, and uterine fibromyoma--in 1 patient. Six members underwent a cytogenetic examination using the metaphase method for peripheral blood lymphocytes and G-banding of chromosomes. Two patients with DPLI carried 8.7 and 16.7% of hyperaneuploid cells and one--20% of cells with double minute chromosomes (DMS). It is suggested that formation and subsequent significant increase in hyperaneuploid and DMS cells could have been responsible for DPLI development in the family. PMID- 12530261 TI - [Effectiveness of high-dose polychemotherapy and autologous hemopoietic cell transplantation in patients with low-grade non-Hodgkin lymphoma]. AB - The study included 70 patients with high-grade malignant non-Hodgkin's lymphoma (Kiel) who received the following treatment: group 1(24)--6-8 cycles of CHOP; group 2 (relapse or tumor progression after standard chemotherapy--13)--6 cycles of CHOP and Dexa-BEAM, and group 3 (relapse or tumor progression after standard chemotherapy--8)--6 cycles of CHOP plus Dexa-BEAM plus autologous hemopoietic cell transplantation with CBV or BEAM conditioning regimen (bone marrow or peripheral stem cells). The groups were comparable as far as gender and age are concerned. Overall 5-year survival rates were 81, 20 and 48%, respectively. CHOP Dexa-BEAM plus autologous hemopoietic cell transplantation proved superior to CHOP-Dexa-BEAM alone. PMID- 12530262 TI - [Genetic polymorphism of steroid 17 alpha-hydroxylase/17,20-lyase (CYP17) and hyperinsulinemia in endometrial carcinoma]. AB - Initiation and/or promotion of endometrial carcinoma is considered to be associated with estrogens and androgens (androstendione) excess as well as hyperinsulinemia and resistance to insulin. It is possible that certain polymorphisms of the genes involved in steroidogenesis or steroid metabolism contribute to carcinoma susceptibility. In the current study, we compared the role of CYP17 biallelic MspA1) polymorphism in 114 endometrial carcinoma patients and 182 healthy women. According to our data, A2/A2 CYP17 genotype traditionally regarded as "unfavorable" was less frequent in cancer patients than in control which confirmed the results of two previous publications. For the first time, carriers of the genotype were shown to have relatively low levels of blood insulin and C-peptide. No significant difference was found between mean concentrations of testosterone, dehydroepiandrosterone sulfate and those of estradiol in the carriers of various CYP17 genotypes with endometrial cancer. Hence, CYP17 polymorphism which is represented by the "normal" A1/A1 genotype might be a factor of risk for endometrial carcinoma. Since this genetic variety may develop through an unconventional (nonsteroid) pathway, taking relevant preventive measures in high-risk groups should be recommended. PMID- 12530263 TI - [Serous-papillary endometrial carcinoma (clinico-morphological features)]. AB - The share of seroso-papillary endometrial carcinoma (SPEC) for the past 4 years has been 6.4%--101 out of 1,567 endometrial cancers. There has been lower incidence of obesity, diabetes mellitus and infertility in cases of SPEC than endometrioid carcinoma (EEC)--18.8 and 53.7; 12.9 and 23.3; 8.8 and 20.6%, respectively, (p < 0.05). Only 60.4% of SPEC patients had locally-advanced tumors (stage I--35.6; stage II--24.8%) while tumor dissemination was reported in 39.6% (stage III--27.7 and stage IV--11.9%). Among EEC patients, locally-advanced tumors were detected in 89.6% (stage I--62.9 and stage II--26.7%) and disseminated tumor incidence was 3 times lower than in SPEC--10.4% (stage III- 8.1 and stage IV--2.3%). Deep invasion into the stroma (more than 10 mm) was registered in 27.7% (SPEC) while in EEC--6.0% (p(0.05). Lymphogenous metastasis was much more common in SPEC (13.9%) as compared with less than 4.1% in EEC patients. There was no correlation between its presence and depth of invasion in the myometrium (p(0.05). High incidence of association of metastasis and superficial or deep invasion was reported for SPEC. This tumor should be classified as pathogenetic variant II of clinico-morphological changes. PMID- 12530264 TI - [Immunohistochemical identification of hormonally-inactive tumors of the adrenal gland]. AB - An immunohistochemical study was carried out to differentiate between such neoplasms of the cortical and cerebral layers of the adrenal as adrenocortical tumor and pheochromocytoma. It used antibodies to chromogranin A, synaptophysin, vimentin, cytokeratin 8, pancytokeratin, carcinoembryonic antigen, epithelial membrane antigen, neurofilaments, alpha-fetoprotein, alpha-antitripsin and neuron specific enolase. Chromogranin A and synaptophysin appeared to be immunohistochemical markers for pheochromocytoma since it responded negatively to vimentin and cytokeratin. Adenocortical tumors were associated with positive reaction to vimentin and a negative one to cytokeratin. These characteristics were used to differentiate adrenocortical tumors from adenomas which responded to cytokeratin and hardly to vimentin. A possibility of adrenal tumors consisting of endocrine and neurogenic cells is considered. PMID- 12530265 TI - [Formalized method of deciding on treatment options in clinical oncology]. AB - A method of constructing a "tree of problem solving" is suggested to work out viable and economical strategies of clinical therapy of tumors. The algorithm is illustrated by solving a hypothetical problem. A clinico-economic index is used to compare different modalities of tumor therapy, its lowest value representing the best regimen. PMID- 12530266 TI - [Comparative pharmaco-economic analysis of docetaxel with cisplatin and cyclophosphamide with cisplatin in first-line chemotherapy of advanced ovarian cancer]. PMID- 12530267 TI - [Susceptibility of wild and knockout p53 FVB/N line mice to benz(a)pyrene-induced subcutaneous sarcoma]. AB - Susceptibility to benz(a)pyrene (BP) has been compared between wild and knockout p53 homo- and heterozygous FVB/N, C57BL/6 and NMRI line mice. Each group included 34-80 animals. Each animal was injected 2 mg BP, s.c., in sunflower oil solution. The animals were followed up for 24 weeks. Fibrosarcomas arose at the site of injection in: wild FVB/N line mice--93%; FVB/N heterozygous--100%; p53 homozygous knockout--95%; C57B1/6--84%, and NMRI mice--83%. Mean latent period was 68, 66, 69, 112 and 109 days, respectively. Hence, FVB/N line mice developed tumors much earlier than C57B1/6 or NMRI did, suggesting their higher susceptibility to carcinogenic BP injection. No significant difference in susceptibility to carcinogen versus p53 expression was reported. PMID- 12530268 TI - [Use of Azopiram for rapid diagnosis of epithelial tumors of the urinary bladder (experimental data)]. AB - Correlation of development of tumor pathology and macro- and microhematuria was investigated in the experiments using 50 male rats with N-butyl-N-(4 hydroxybutyl)-nitrosoamine-induced tumors of the urinary bladder. Our highly sensitive agent Azopiram was employed to monitor hematuria. Microhematuria, which subsequently developed into macrohematuria as tumor matured and turned malignant, was identified as early as at such incipient neoplastic changes as focal hyperplasia of the urothelium and micropapillomatosis. The test can be used outside clinic as a measure of self-control in recurrent cancer of the urinary bladder. PMID- 12530269 TI - [Experimental evaluation of immunotoxicity of olipifat]. AB - The paper presents our data on the influence of Olipifat on the mass and cell patterns of the immune system organs, phagocytic activity of macrophages, number of antibody-producing B-lymphocytes and immune rosette-forming T-lymphocytes. Olipifat showed no immunotoxic characteristics; it stimulated T-system immunity as evidenced by a significant increase in the number of immune rosette-forming T lymphocytes in mice after injection of 100 or 50 mg/kg. PMID- 12530270 TI - [Biotherapy of experimental tumors with vaccines of Venezuelan encephalomyelitis virus, parotitis and influenza viruses and cytologic evaluation of effectiveness]. AB - Inhibition of experimental tumor growth, life-span, mitotic activity and cytologic damage have been studied in cells of ascitic and solid carcinoma of Ehrlich in the course of biotherapy with vaccines of Venezuelan encephalomyelitis virus, mumps and influenza viruses. Summing up, it is concluded that the vaccine of Venezuelan encephalomyelitis virus looks most promising as far as further research in development of an antitumor viral drug is concerned. PMID- 12530271 TI - [Experience with glyciphonic ointment therapy for primary multiple basal cell carcinoma of the skin]. AB - The effectiveness of glycifone ointment treatment for primary multiple basal cell carcinoma of the skin was evaluated in 28 patients. 27 cases were cured following one or two courses the duration of which was determined by the number of foci, pattern of tumor and its total surface area. PMID- 12530272 TI - [Use of Doppler sonography in the diagnosis of soft tissue malignancies]. PMID- 12530273 TI - [The dynamics of psychologic distress in patients with maxillofacial and laryngeal malignancies]. AB - The study of psychological dynamics involved in malignancies of the maxillofacial area and larynx included 38 male patients (tumors in maxillofacial area--26; neoplasia of larynx--12). Patients were given different regimens of radiation and radiotherapy; 11 were operated on. Psycho-disorders induced by tumor disease and admission to cancer ward were reported in 35 (92%). With time, such disorders were progressively overshadowed by somato-psychic changes, with asthenic syndrome prevailing, and subsequently psychopath-like features developed. Psycho-organic syndrome was registered in 16 patients as they were approaching terminal stage. PMID- 12530274 TI - [Neck tumors: possible reasons for radiologic misdiagnosis]. AB - Neck tumors may be caused by a variety of both inflammatory and non-inflammatory lesions in organs and tissues. US and CT are successfully used to detect neck pathology but misdiagnosis still occurs. To analyze the situation, the data on 124 patients with tumors, cysts, inflammatory lesions and metastasis (after surgery and radiotherapy) were evaluated. The two categories of factors were identified: (1) poor quality of US and CT images due to anatomical peculiarities, thick layers of subcutaneous fatty tissue, deep location of lesions, and scars and edema produced by previous surgery and radiotherapy. As far as CT is concerned, imperfections may be due to artifacts and insignificant difference between tumor and normal tissue densities; (2) misinterpretation of images due to tumor and salivary gland merging to form one block, similarity of outlines of malignant and benign tumors, cysts and cyst-like metastases, neoplastic and inflammatory lesions. The ways of improving the quality of US and CT images are suggested. In particular, FNAB should be used to differentiate between malignant and benign tumors. PMID- 12530275 TI - [Clinical and radiologic signs of pulmonary and pleural lesions in childhood Hodgkin's disease]. AB - Pulmonary and pleural involvement was identified in 17% (42.2% at the debut of the disease, while 57.8%--in the course of tumor progression) out of 264 children with Hodgkin's disease. Basing on X-ray evidence, pulmonary lesions were classified as perinodal, focal-infiltrative, disseminated, nodular, mixed infiltration and lobitis, while those of the pleura--exudative pleurisy and tumor masses involving the pleura. Focal-infiltrative pattern was most common (53%). The best results were reported in the treatment of perinodal infiltration, whereas the worst--disseminated and mixed infiltration as well as pleural lesions. PMID- 12530276 TI - [Quantitative evaluation of physical vitality and quality of life in combined therapy of patients with disseminated breast cancer]. AB - The paper discusses the criteria of quantitative presentation of data on individual sensitivities and the body's vital systems potential used in treatment of cancer patients. An index is suggested to assess damage done to a system using an "automated classifier". PMID- 12530277 TI - [Effect of small doses of ionizing radiation on selected antioxidant and immunologic parameters in patients with cervical cancer]. AB - We analyzed the indices of antioxidant and immune systems in 69 patients with cervical carcinoma T3NxMo at different stages of radiotherapy. Radiation doses of 10-20 Gy were followed by certain normalization of antioxidant enzyme levels, increased concentrations of vitamins E and A in immunocompetent blood cells and improved the penetration and elastic properties of formed cells. The dosage stimulated the activity of helper-inducing and cytotoxic T-cells. Such effects should be taken into account when working out regimens of complex treatment of locally-advanced cervical carcinoma which include cytostatic drugs and radiation. PMID- 12530278 TI - [Mediastinal Hodgkin's disease and non-Hodgkin lymphoma in childhood]. AB - A retrospective evaluation was carried out of the data on 319 children, aged up to 15, with mediastinal Hodgkin's disease, treated at Petrov Research Institute of Oncology (1973-1999). Non-Hodgkin's lymphoma was diagnosed in 113 patients (35.5%); Hodgkin's disease--205 (64.5%). Correlations were established between certain patterns of disease, on the one hand, and age, duration of disease, clinical and hematological features, on the other. Non-Hodgkin's lymphoma was relatively more aggressive. PMID- 12530279 TI - Lead-associated deficits in stature, mental ability and behaviour in children in Karachi. AB - This study was conducted to evaluate the cumulative and steady-state lead burden in children from Karachi, an area of high lead exposure, and to assess the degree of damage to physical growth and mental ability related to lead exposure. A cross sectional survey was conducted in seven primary schools from around Karachi. Shed primary teeth and blood samples were collected from students of grades I to III (age range 6-10 years) and were analysed for lead by atomic absorption spectrophotometry. Haemoglobin concentration, height, weight and head circumference were measured. IQ was estimated using Raven's Standard Progressive Matrices. Classroom behaviour was rated by teachers and school performance was estimated from the percentage mark in a school examination taken just before the date of the IQ test. Complete data were available for 138 children. Over 80% of children had lead levels above the safety limit set by the United States Centers for Disease Control and Prevention. The blood lead levels in boys did not differ significantly from that in girls. Significant differences were observed between the schools. Univariate analysis showed negative association of blood lead with haemoglobin, IQ and height. Tooth lead was negatively associated with height, classroom behaviour and performance. When adjusted for other confounding variables, blood lead was negatively associated with haemoglobin and IQ, whereas tooth lead was negatively associated only with classroom behaviour. Height was negatively associated with blood or tooth lead. These results were further supported when the upper and lower quintiles for blood or tooth lead were compared. Tooth lead level was not a better marker of lead poisoning than blood lead level in our study population. These data demonstrate the association of increased lead with impaired learning and adverse behaviour in Karachi children and call for strict government regulations to limit environmental lead burden. PMID- 12530280 TI - Effect of zinc on the tuberculin response of children exposed to adults with smear-positive tuberculosis. AB - The tuberculin test (PPD) is used frequently in the diagnosis of tuberculosis. PPD, however, relies on an intact cell-mediated immunity and infected children often have false negative results. This study assessed whether a single oral zinc supplement modifies the PPD induration size and its association with nutritional status in Brazilian children. Ninety-eight children below 15 years of age who had been exposed to adults with smear-positive pulmonary TB in 1998 were tested by PPD in 1998 and 2000. Children were randomised in 2000 to receive a single oral dose of zinc sulphate or a placebo at the time of administering the PPD. Forty three (44%) children were PPD-positive in 1998 and 54 (55%) in 2000. A higher proportion of children were classified as PPD-positive in 2000 in the zinc supplemented group (57.1%) than in the placebo group (53.1%). PPD indurations were larger in children receiving zinc (mean 18.5 and 15.5 mm in the zinc and placebo groups, respectively) (p < 0.03). Mean induration sizes in 2000 were larger in zinc-supplemented children, regardless of their nutritional status. Our study demonstrates that zinc increases the PPD induration size in children irrespective of nutritional state. Zinc supplementation could work by correcting asymptomatic or marginal zinc deficiencies or as a non-specific booster of immunological mechanisms (whether or not there is a deficiency). PMID- 12530281 TI - Serum and breast-milk vitamin A in women during lactation in rural Chiang Mai, Thailand. AB - Vitamin A deficiency can occur during lactation and breast-milk vitamin A has been recommended for monitoring the vitamin A status of lactating women and their infants. This study aimed to investigate the vitamin A status of lactating women in relation to race, age, parity, duration of lactation and anthropometric status. A cross-sectional study was conducted among 262 lactating women in rural Chiang Mai, Thailand. Blood and breast-milk samples were collected. Serum retinol, carotene and breast-milk retinol concentrations were analysed by high performance liquid chromatography. The results show that mean serum retinol and breast-milk retinol in hill tribes were significantly lower than in Thais, 1.91 (0.59) and 0.79 (0.52) compared with 2.10 (0.51) and 1.04 (0.58) mumol/L, respectively. Mean serum retinol and breast-milk retinol were highest during the 1st 3 months of lactation. Maternal age, parity and anthropometric status (BMI) were not associated with serum or breast-milk retinol concentrations. There was a significant relationship between serum and breast-milk retinol values in women who breastfed for 6 months or longer (regression co-efficients 0.30; 95% CI 0.16, 0.43). Breast-milk retinol levels declined significantly from 4 to 12 months after delivery, which could increase the risk of vitamin A deficiency in children who were exclusively breastfed or receiving inappropriate complementary foods during this period. Weaning foods which commence at 6 months and have an adequate vitamin A content should ensure that the vitamin A status of the young child is maintained. PMID- 12530282 TI - The transplacental transfer of IgG subclasses: influence of prematurity and low birthweight in the Gambian population. AB - Having established previously that prematurity in The Gambia is associated with impaired maternofetal transfer of some specific antibodies, we investigated the influence of low birthweight and prematurity on placental transfer of IgG subclasses in 180 Gambian mothers and their newborn babies. A physician-blinded, cross-sectional study of 180 mother-baby pairs was carried out in the labour ward of Bansang Hospital, The Gambia. Paired maternal and cord serum samples were collected from mothers and their newborns. Serum IgG subclass levels were measured by laser nephelometry. Placental transfer of all IgG subclasses was significantly reduced in preterm newborns compared with term newborns (p < 0.01). In low birthweight (LBW) babies, the transfer of IgG1 (p = 0.03) and IgG2 (p < 0.01) subclasses, but not IgG3 and IgG4, was significantly reduced. In preterm neonates with an adequate birthweight and term neonates with a low birthweight, all IgG subclasses were transferred with reduced efficiency, but IgG1 and IgG2 subclasses were transferred with significantly less efficiency than IgG3 and IgG4. These results confirm that prematurity and low birthweight are associated with impaired placental transfer of IgG1 and IgG2 subclasses in this Gambian population. This impairment might explain the susceptibility of these newborns to certain polysaccharide-encapsulated organisms. PMID- 12530283 TI - Neurocysticercosis in children: clinical characteristics and outcome. AB - A study was undertaken on 176 children with neurocysticercosis to determine the clinical behaviour and long-term outcome of cases. The children were followed up prospectively in the paediatric neurology clinic for 6 years. Diagnosis was based primarily on CT scans. All the CT lesions were parenchymal and mostly single (87%) with ring enhancement and peri-lesional oedema. Partial seizure was the predominant presenting seizure type. About 65% of cases had recurrences at varying intervals during treatment. Albendazole therapy given to some cases did not appear to have any beneficial effect on seizure control. Repeat CTs done at varying intervals after the first seizures showed disappearance of the lesions in about 50% of cases. The majority of lesions disappeared without the use of albendazole. Recurrence of seizures after withdrawal of anti-epileptic medication was observed in 19% of the cases. Parenchymal neurocysticercosis in children commonly presents with partial seizures and ring lesions on CT. The disease can be managed well by anti-epileptic medication alone and the prognosis is good. PMID- 12530284 TI - Seroepidemiology of varicella-zoster virus in Bangladesh. AB - Data on the seroprevalence of antibodies protective against the varicella-zoster virus are needed to develop strategies to prevent varicella infections in Bangladesh. Of 1209 patients evaluated at referral-level health facilities in Dhaka, 943 (78%) had no known history of chickenpox and were tested by latex agglutination for the presence of varicella-zoster antibody in serum. Forty-one per cent (386) of the 943 specimens tested were negative. Seropositivity was highest among neonates (83%), declined sharply to 19% in those aged 7-12 months, and thereafter rose steadily with age until a plateau of 85% was reached after the age of 16 years. This first report of varicella-zoster antibody seroprevalence in Bangladesh suggests that, as in other tropical areas, a significant proportion of children, adolescents and adults are susceptible. Children aged from 15 months to early adolescence might be the most important group to target with the vaccine currently available. However, to ensure successful immunisation, further, population-based seroprevalence data are needed, as are an assessment of the vaccine's acceptability and the accessibility of the target population. Incomplete coverage of young children could result in delayed acquisition, and, ultimately, in more severe disease. PMID- 12530285 TI - Evaluation of polymerase chain reaction (PCR) for diagnosing Haemophilus influenzae b meningitis. AB - A polymerase chain reaction (PCR) for detecting Hib in cerebrospinal fluid (CSF) was evaluated and compared with culture and a latex agglutination test (LAT) in a hospital-based prospective surveillance. We studied 107 children aged from 1 month to 12 years with a clinical and CSF profile suggestive of acute bacterial meningitis. CSF culture was performed on blood-chocolate agar by standard technique, LAT by a commercially available kit (Wellcogen) and PCR using total DNA extracted from CSF samples. Of 107 children, 79% had received one or more doses of injectable antibiotics. Hib was detected by culture in 14 cases, by LAT in 23 and by PCR in 37. All CSF samples that reveal Hib by culture or LAT had a PCR positive for Hib (sensitivity 100%). PCR also detected 14 additional cases of Hib meningitis which were not detected by culture or LAT. We conclude that PCR is a sensitive and specific diagnostic tool that may be valuable in a population with high pre-hospital antibiotic usage. PMID- 12530286 TI - A new approach for neurological evaluation of infants in resource-poor settings. AB - Research assessing the neurological development of infants in developing countries is scanty as no suitable standardised tests are available for field-use in constrained circumstances. We describe the development and application of two simple assessments. Firstly, we aimed to develop a test suitable for assessing acute neurological disturbances caused by such diverse effects as infections, drugs or toxins. This test (Shoklo Neurological Test) is aimed at infants between 9 and 36 months. The second test (Shoklo Developmental Test) is aimed not only to follow the evolution of the signs tested initially in the acute phase but also to evaluate later neurodevelopmental sequelae which might be caused by the same events. The latter test is suitable for infants aged from 3 to 12 months. Both tests can be performed easily in non-optimal conditions. The examinations were tested in a cohort of infants from a Karen refugee camp and administered in a rural setting by health workers, after appropriate training. In order to validate the tests we also applied them to a cohort of London infants. The Griffiths Developmental Scales were applied in the same infants and both the Shoklo Neurological and the Shoklo Developmental Tests showed good correlation with this standardised neurodevelopmental assessment. PMID- 12530287 TI - Epidemiology of Burkitt's lymphoma in Enugu, Nigeria. AB - Forty-four children between 3 and 14 years of age with Burkitt's lymphoma were studied at the University of Nigeria Teaching Hospital, Enugu, Nigeria from November 1991 to March 1995. Mean age at presentation was 7.6 years with a male:female ratio of 2.7:1. Eighty-six per cent of the patients lived in a rural area. Of the 23 (52%) from Enugu State, 16 were living in contiguous local government areas with ten clustered round the middle of the year. All the children were of the lowest socio-economic class, 75% of them being in class V. The incidence of Burkitt's lymphoma was higher during the dry season and the period of high malaria transmission (p < 0.05). Abdominal tumour was the commonest mode of presentation (32% of patients). Thirty-six patients were treated with cyclophosphamide, Oncovin, methotrexate and predinisolone (COMP), five received cyclophosphamide alone and three had cyclophosphamide, methotrexate and prednisolone (CMP). Multi-drug regimens achieved better results which were significantly unrelated to the number of cycles of therapy received. Overall, 48% had complete and 35% partial remission, 23% relapsed and 16 (36%) patients died. The epidemiological characteristics of Burkitt's lymphoma in this study were similar to those in other tropical regions. PMID- 12530288 TI - Normal thyroid volume by ultrasonography in healthy children. AB - With the objective of creating standards for the volume of the thyroid gland by ultrasonography in the 0-16-year age group, thyroid volumes of 302 healthy children (150 boys, 152 girls) were measured by ultrasonography. The transverse (x), sagittal (y) and anteroposterior (z) lengths of the right and left lobes and isthmus were measured. Volumes of these were calculated by the ellipsoid volume formula [V = (pi/6) x x x y x z]. Statistical analysis was done using the Kruskal Wallis and Student t tests. The cases were divided into six and eight groups by age and height, respectively. There was a significant positive correlation between thyroid volume, age, height and weight. Thyroid volumes in children of 12 years and over were significantly different from those in the younger age groups. PMID- 12530289 TI - Isolated splenic infarction owing to group B Salmonella: case report. AB - The clinical spectrum of extra-intestinal salmonellosis, comprising enteric fever and invasive infections owing to non-typhoidal Salmonellae, is well known. We report an otherwise healthy patient with isolated splenic infarction caused by group B Salmonella. She was seropositive for the O antigen of Salmonella group B and stool cultures were positive for group B Salmonellae. After appropriate antimicrobial therapy, her complaints disappeared and microbiological tests for Salmonellae became negative. PMID- 12530290 TI - The first case report in Saudi Arabia of diabetes mellitus and cystic fibrosis in two siblings. PMID- 12530291 TI - Managing injectable drugs: an evolving challenge for pharmacy benefit managers. PMID- 12530292 TI - Improving the efficiency of disease management programs by assessing patients' readiness to change. PMID- 12530293 TI - Promoting physical activity: a profile of health plan programs and initiatives. AB - A study was conducted to capture information on health plans' initiatives to promote physical activity among members and in community settings. This investigation was based on a descriptive nationwide study of American Association of Health Plans- and Blue Cross and Blue Shield Association-member plans conducted from October 2001 to February 2002. Sixty individuals with responsibility for health-promotion programs were surveyed, and 24 provided in depth interviews. The measures used in this evaluation were the number and type of member- and community-based physical activity programs/initiatives offered by health plans, and the types of programs evaluated. Information was captured on more than 373 physical activity initiatives. Thirty-five percent of surveyed health plans responded, representing more than 62 million health plan members; the participation rate for in-depth interviews was 80%. Most health plans integrate physical activity messages into routine services for members (92%) and sponsor community races/walks and health fairs (85%). Physical activity programs are offered to improve member health (92%), increase member satisfaction (80%), and reduce long-term health care costs (62%). Few rigorous evaluations have been conducted on these programs, and return-on-investment data are scarce. Although some health plans currently play a relatively active role in promoting physical fitness to their members and the community, multiple opportunities exist for health plan involvement in various settings with different populations. PMID- 12530294 TI - Vision care benefits and outcomes of treatment of keratoconus. AB - Coverage for basic vision care services is offered by a number of national and regional vision care networks. When eyewear is covered, cosmetic contact lenses are usually provided in lieu of glasses. There are a few disorders for which contact lenses are prescribed for a "medically necessary" condition; keratoconus is an example. Some plans provide full benefits for noncosmetic contact lenses for the treatment of keratoconus, and outcomes measures have demonstrated that visual improvement has been excellent. Evidence of the vision benefit offered to those with keratoconus is found in patient-satisfaction surveys, which provide self-reports of improvements in near and distance vision. PMID- 12530295 TI - Using IT to enhance patient safety. PMID- 12530296 TI - ["Morphology" journal today]. PMID- 12530297 TI - [The pride of the St. Petersburg University--Aleksandr Stanislavovich Dogel']. PMID- 12530298 TI - [The Tomsk period of professor A. S. Dogel's work]. PMID- 12530299 TI - [Main directions of research development in experimental histology and cytology in the third millennium]. PMID- 12530300 TI - [The assessment of structure-function interrelationship from the standpoint of functional systems]. AB - One of the major methodological problems of morphology--the correlation of "structure" and "function" categories in normal anatomy and histology--is discussed. The issues of mobility of concepts concerning organization of functional systems, differon tissue organization, tissue classification including the special place of extraembryonic organs, are examined. The organizational matters associated with the introduction of a new speciality 03.00.25--histology, cytology, cell biology--and with the necessity of revision of the program of qualifying examination in philosophy for the candidate's degree in the field of histology, are discussed. PMID- 12530301 TI - [Morphological peculiarities of rat liver reaction to chronic xenobiotic exposure]. AB - The data of the morphological studies of liver of animals subjected to chronic (3 months long) intoxication with different chemical substances (ammonium nitrogen, nitrite nitrogen, fluorine, chromium, arsenic) are presented. The xenobiotics tested induced stereotyped responses in liver which differed in their intensity and the extent of hemomicrocirculatory and inflammatory-destructive alterations. Most pronounced reactive changes were found in animals exposed to arsenic compounds, while sodium nitrate caused practically no destructive changes in hepatocytes. PMID- 12530302 TI - [Peculiarities of the structure of the speech-motor cortex of the brain frontal lobe in a deaf-mute child]. AB - The aim of present investigation was the study of the cytoarchitecture of the motor-speech areas 44 and 45 of the brain cortex of a 4-year-old deaf-mute child. We have also compared cytoarchitecture of the motor-speech zone of the brain cortex in a deaf-mute child with the same brain region in a normal child of a similar age. Series of 20 mm-thick frontal sections stained with cresyl violet were studied using morphometric methods. Our data indicate the significant anomalies of both the brain gross structure and cytoarchitecture of the motor speech zone of the brain cortex in a deaf-mute child. Considerable decrease in the values of neuronal profile area in layers III3 and V and in satellite gliocyte density was observed while the neuro-glial ratios were changed. Our results certify that development of child's brain is disturbed in conditions of speech dysfunction and auditory deficiency. PMID- 12530304 TI - [Formation of neocortex in rats after prenatal hypoxia]. AB - Exposure of pregnant rats to the atmosphere containing 7.56% of oxygen for one hour was found to cause the disturbances in the nerve system development in their offspring. The exposure to acute hypoxia at days 13 or 16 of embryonic development induced an attenuation and underdevelopment of brain cortical layers, disturbances in cell orientation and differentiation, i.e. caused the modifications of basic histogenetic processes that were active during this period, including proliferation, migration and differentiation, thus resulting in the changes of structural characteristics of neocortical layers. The effect of hypoxia during the later fetal period--at day 19 of embryonic development, when cell proliferation in the brain ceases while the processes of differentiation are enhanced,--is less damaging. The cortical layers formed in experimental animals are not significantly different from those in control rats, however the death of individual neurons and gliocytes takes place. PMID- 12530303 TI - [Bcl-2 protein expression in developing human brain]. AB - Using immunocytochemical methods, the expression of bcl-2 antiapoptotic protein was studied in developing brain of human 5-8-week embryos. It was established that during the period studied several zones of bcl-2-positive cell concentration were present. These included the ventricular zone of the brain vesicles wall, cortical plate, medullary nuclei, vascular plexus primordium (in 5-6-week embryos), IV ventricle roof and vascular plexus of rhombencephalon. In the area of forming leptomeninx, bcl-2 protein was demonstrated in the endothelium of blood vessels. The detection of intensified expression of bcl-2 protein in some cellular populations of developing human brain proves their increased stability against apoptosis and indicates their priority significance in the provision of normal neurohistogenesis. PMID- 12530305 TI - [Structural changes in dendritic spines of the pyramidal neurons of layer III of the rat sensory-motor cortex during remote postischemic period]. AB - The combination of Golgi method with the lipid phosphorilate mark Dil (1,1' dioctadecyl-3,3,3 cents, 3 cents-tetramethyl-indocarbocyanine perchlorate) and confocal laser scanning microscope for demonstration of the structure of dendritic tree and dendritic spines enables exact determination of spatial organization of small dendrites and their spines at significant distances in norm and especially in postischemic period. The regularities of dendritic spine reorganization in layer III pyramidal neurons of the cerebral sensory-motor cortex during 9 months after short-term (10 min) total brain ischemia were established. PMID- 12530306 TI - [Morpho-functional changes of Mauthner neurons after treatment with neuropeptide kyotorphin ]. AB - Influence of neuropeptide kyotorphin (L-thyrosyl-L-arginine) treatment on goldfish Mauthner cells was studied separately or in combination with prolonged natural stimulation of these neurons. The neuroprotective effect of this substance at both functional and ultrastructural levels was demonstrated. Possible mechanisms of kyotorphin action is discussed with regard to its modulating effect on intracellular calcium ion concentration. PMID- 12530307 TI - [Comparative morphological analysis of neuron populations in ganglion layer of rodent cerebellar cortex]. AB - The characteristics of basophilic substance distribution, linear dimensions and Purkinje cell (PC) density per unit of gyrus length in ganglion cell layer of cerebellar cortex were studied in ecological-morphological groups of small burrow rodents. Polymorphism specificity of neuron populations in ganglion cell layer was demonstrated in regard to degree of chromophilia of neuron cytoplasm which is characterized by obvious prevalence of normo- and hyperchromatic cells in all PC populations. The correlation between average values of PC body profile area, density of neurons distribution in cell population and the degree of animal adaptation to their environment was analyzed. PMID- 12530308 TI - [Axonal reaction precedes demyelination in experimental models of multiple sclerosis]. AB - Taking into consideration that myelin phospholipids may be partially synthesized in neuronal bodies, while neurilemma readily reacts with antibodies against gangliosides by changing the properties of membrane ionic channels, the attempt was made to test the proposed assumption of the early axonal reaction in demyelinating processes in experimental models of multiple sclerosis. The models of chronic allergic encephalomyelitis in Lewis rats injected with homogenate of highly purified myelin or total brain gangliosides were used. First signs of demyelination (the destruction of intermediate dense lines) were demonstrated in the inner layers of myelin close to axon and were shown to develop synchronously with the aggregation of filamentous-tubular material in the neuroplasm. These changes are associated with significant shift of the ratio of myelin sheath thickness to axonal diameter (from 1:7-1:3 to 2:1-3:1). This swelling of myelin seems to be caused by neuroplasmic proteins aggregation, that must be accompanied by the drop in oncotic pressure and the separation of loosely-bound water fraction that may be assimilated by myelin. At light microscopic level the increase of myelin thickness is clearly observed that is in exact correspondence with the decrease in axonal diameter. The process starts with the exfoliation and swelling of the nodes of Ranvier and the incisures of myelin, which fuse after elongation, that corresponds to the total disintegration of myelin with the preservation of continuity of axon which appears to be harshly shrunken. PMID- 12530309 TI - [Morphological aspects of heart remodeling in chronic heart failure]. AB - Intraoperative biopsy specimens of the right atrium auricula from patients at different stages of chronic heart failure (CHF) were studied using the methods of light and electron microscopy. Marked heteromorphism of cardiomyocytes (CMC) was demonstrated which was found to be correlated to CHF progression. CMC showing the signs of hypertrophy and organelle hyperplasia, sarcomere discomplexation, myofibril fragmentation were identified, as well as CMC containing numerous pinocytotic vesicles, caveolae, distended T-system tubules. Apoptotic CMC and interstitial cells were also shown. No significant correlation between the myocardial cell apoptosis and CHF stage was established. PMID- 12530310 TI - [Lymphoid structures of human duodenum in adolescent and juvenile age]. AB - Using morphometric methods, the microtopography and quantitative proportions of lymphoid cells in structural components of lymphoid tissue were studied in the duodenal wall of the persons of adolescent and juvenile age. In adolescence, in contrast to juvenile age, the intestinal wall was found to contain all the morphological structures, characteristic to functional activity of lymphoid tissue (lymph nodules with and without the germinal centers). In adolescence numerous cells in mitosis and immature cells were found in intestinal lymphoid structures, while no mitotic figures were detected in the similar structures in juvenile age. Duodenal lamina propria contained more antigen-producing plasma cells in adolescence than in youth. This complex of cell correlations is indicative of enhanced local immune response in adolescence and may be explained by an age-dependent active neuro-hormonal body reconstruction. PMID- 12530311 TI - [Structure of the adrenal gland in rat strain with hereditary stress-induced arterial hypertension nursed by Wistar dams]. AB - The comparative investigation of the adrenal structure in two groups of rats of Inherited Stress-Induced Arterial Hypertension (ISIAH) strain was conducted. The animals of the first group were nursed by normotensive Wistar rats, while those of the second (control) group were reared by their own mothers. The volume of the adrenal medulla in rats of the first group was found to exceed that in the second group. In the adrenal zona glomerulosa and zona fasciculata of 3-week-old rats of the first group parenchymal-stromal ratio and the average volume of adrenocorticocytes were lower than those in the animals of the second group. This was accompanied by a decrease in mitochondrial volume density and accumulation of lipid droplets in the cytoplasm, indicating a reduction of hormone-producing activity of endocrinocytes in the animals the first group as compared to controls. By 6 month of age, arterial pressure and quantitative parameters of adrenal medulla and cortex in the animals the first group were..... as compared to those in the second group. It is suggested that the nursing of ISIAH rat pups by normotensive Wistar dams had modulating effect on the adrenal structure and therefore on arterial hypertension development. PMID- 12530312 TI - [Quantitative assessment of the processes of insulin synthesis and secretion in B endocrinocytes of pancreatic islets]. AB - Immunofluorescent methods allowing the detection of different hormones in endocrine cells, are widely used in many works, however, their application is usually limited to the description of qualitative criteria, while the reactions of indirect immunofluorescence permit their quantitative assessment. The purpose of the present work was the definition of the criteria for the quantitative assessment of the activities of the processes of insulin synthesis and secretion in pancreatic B-endocrinocytes by examination of changes of their fluorescence using classical activation of these cells by glucose. The investigations performed indicate that the main parameters of activity of insulin synthesis and secretion in pancreatic B-endocrinocytes are the area occupied by immunoreactive material, its concentration and heterogeneity. As the secretory response of pancreatic B-endocrinocytes is of variable intensity, the classification of these cells into groups with different activity of the processes of insulin synthesis and secretion is proposed. This approach enables the quantitative assessment of the action of different insulotropic agents, opening promising prospects for their further investigation. PMID- 12530313 TI - [Morphological aspects of dermatotrophic action of methyluracil applied epicutaneously]. AB - The changes of the histological structure of intact skin and of the skin within the area of burn trauma, caused by treatment with methyluracil-containing ointments, were studied. Morphometric investigations employing computer technologies have been performed using "DiaMorph" device-program complex (Russia). It was established that methyluracil possessed wound-healing properties by stimulating epidermal proliferation in the zone of injury and by modifying an inflammatory reaction enhancing its fibroblastic component. Daily applications of methyluracil to intact skin for a period of half a year resulted in some reduction of reserve capabilities of cambial keratinocytes, that was objectively manifested by a decrease of epidermal germinal layer thickness. PMID- 12530314 TI - [Ultrastructural organization of the human lamellar bone tissue mineral component in aged and elderly]. AB - The lamellar bone mineral component in mature and senile age was studied using the methods of transmission electron microscopy and cryofractography. This component was shown to be formed by crystals (13-4 x 7-8 x 3-4 nm) of the prismatic shape that formed coplanar units (29-3 x 18-21 x 3-4 nm). The peculiarities of mineral particle distribution inside and between the collagen fibrils, as well as on the completely formed areas of bone surface, were detected. It was established that the length and the width of coplanar crystal aggregates was significantly higher in senile age as compared to similar linear parameters defined in persons of mature age (by 17-20 and 5-9%, respectively). PMID- 12530315 TI - [Mitogenic effect and acceleration of stabilization process in the renal glomeruli of hypertrophied kidney after exposure to magnetic field]. AB - Experiments designed to investigate the effects of magnetic fields of different nature, induction, exposure duration and multiplicity, have established a stimulating action of alternating magnetic field of 15 mT applied 2-5 times for 30 min daily on mitotic division of proximal tubule epitheliocytes. The degree of renal glomeruli capillary lumina distention (CLD), defined as the ratio of the total area of glomerular capillaries to glomerular area, was found to be variable, indicating the asynchronism of their filling with blood. Elevated filtration volume in a single kidney left after nephrectomy, was provided by a synchronization of blood filling of renal glomerular capillaries, that was manifested by the shift of their major part to the groups with high CLD and the appearance of glomeruli with extremely dilated capillaries. These changes were more significant and were detected earlier following exposure to alternating magnetic field. PMID- 12530316 TI - [Somatometric characteristics of normal young children and children with endocrine pathology]. AB - Somatometry and somatotyping was performed in 570 healthy children in the first childhood period, attending preschool institutions, and in 78 children with pathology (including 59 children with diabetes mellitus and 19 with thyroid pathology) undergoing clinical investigation and treatment in the Department of Endocrinology of Rostov-on-Don Scientific Research Institute of Obstetrics and Pediatrics. The results obtained revealed that thyroid hyperplasia was most frequent in children of micromuscular type with the average development of bone and adipose somatic components. Congenital hypothyroidism was prevalent in children of macrocorpulent type having the average and high bone mass parameters. In children with diabetes mellitus low values were found for the indices characterizing the degree of development of all anatomical somatic components. PMID- 12530317 TI - [Contribution of outstanding Bulgarian neuromorphologist professor Emilia Zaprianova to the development of the neuromorphology on the verge of the XX-XXI centuries]. PMID- 12530319 TI - Derangements of coagulation and fibrinolysis in infectious diseases. PMID- 12530318 TI - Complement and infectious diseases. PMID- 12530320 TI - The role of chemokines in sepsis and septic shock. PMID- 12530321 TI - Modulation of membrane traffic by microorganisms. PMID- 12530322 TI - The role of neutrophilic secretion products in infectious diseases. PMID- 12530323 TI - Modulation of the vascular endothelium during infection--the role of NF-kappa B activation. PMID- 12530324 TI - Antimicrobial peptides as effector molecules of mammalian host defense. PMID- 12530325 TI - Role of toll-like receptors in response to bacterial infection. PMID- 12530326 TI - CD1 antigen presentation and infectious disease. AB - Taken together, the data generated thus far strongly suggest that CD1 plays a role in the immune response against various infections (table 1). For obvious reasons, the data gathered thus far using model infection systems have focused primarily on the mouse and therefore only examine the role of CD1d. This leaves an important gap in our understanding of the CD1 antigen presentation pathway given the potential role of CD1a, CD1b and CD1c for contributing to antimicrobial immunity. The functional dichotomy between group 1 and group 2 CD1 isoforms obviously requires further analysis. However, we propose that the group 1 CD1 (CD1a, CD1b, CD1c) antigen presentation pathway is closer to the traditional adaptive immune response mechanisms with the capacity to present unique foreign antigens to specific T cells. This broadens the universe antigens that T cells can use to target pathogens and provides important antimicrobial effector mechanisms that may be critical for combating some types of infections. Lipid antigens may also provide a more effective means of targeting intracellular pathogens by T cells since CD1 is able to sample almost all of the intracellular reservoirs that are exploited by this class of pathogen and may provide an important component of the cytotoxic T cell response [80]. On the other hand, the group 2 CD1 protein (CD1d) may be more intermediate in terms of lying functionally between the innate and adaptive immune systems. The activation of CD1d-restricted T cells may, therefore, help bridge the temporal gap between the onset of innate immunity and the purely adaptive responses typified by the MHC restricted T cells. Hence, the CD1d-restricted [table: see text] T cells are primed for rapid high-level cytokine release. In addition, the interaction of CD1d-restricted T cells with CD1d on DCs can trigger the release of IL-4 and GM CSF to promote maturation of tissue-resident DC at the site of infection. The maturation of tissue DC would lead to migration of the activated DC to regional lymph nodes and initiation of MHC-restricted T cell responses. Subsequent IL-12 production by the DC in response to CD1d-mediated T cell stimulation could then drive IFN-gamma production by CD1d-restricted T cells and influence the polarization of the T cell response to infection. In addition, early bursts of IFN-gamma by CD1d-restricted T cells could also upregulate antimicrobial activity in macrophages and activate other important effector cells such as NK cells prior to MHC-restricted T cell responses. In the constant struggle between the microbial pathogen and its host, the evolutionary balance almost always favors the microbe. The rapid rate of evolution and adaptation of the microbe accounts for most of this advantage. Hence, it is not surprising that the host immune system has evolved a complex set of pathways, in addition to the MHC, that are able to recognize and target the unique molecular signatures of infectious microorganisms. The lipid antigens presented by CD1 add to this array and thus provide a further layer of immune defense to the host for combating pathogens. PMID- 12530327 TI - Molecular and cellular mechanisms of bacterial entry into host cells. PMID- 12530328 TI - The role of chemokine receptors in the promotion of viral infections. PMID- 12530329 TI - Virally encoded chemokines and chemokine receptors in the role of viral infections. AB - Large DNA viruses such as pox- and in particular herpesviruses are notorious in their ability to evade the immune system and to be maintained in the general population. Based on the accumulated knowledge reviewed in this study it is evident that important mechanisms of these actions are the acquisition and modification of host-encoded chemokines and chemokine receptors. The described viral molecules leave nothing to chance and have thoroughly and efficiently corrupted the host immune system. Through this process viruses have identified key molecules in antiviral responses by their inhibition of these or potent ways to alter an efficient antiviral response to a weak Th2-driven response. Examples here are the chemokine scavenging by US28, attractance of Th2 cells and regulatory cells by vMIP1-3 and the selective engaging of CCR8 by MC148. Important insights into viral pathology and possible targets for antiviral therapies have been provided by UL33, UL78 and in particular ORF74 and the chances are that many more will follow. In HHV8 vMIP-2 and the chemokine-binding proteins potent anti-inflammatory agents have been provided. These have already had their potential demonstrated in animal models and may in their native or modified forms represent useful therapies in humans. PMID- 12530330 TI - Television images and psychological symptoms after the September 11 terrorist attacks. AB - Exposure to graphic television images may exacerbate psychological symptoms in disaster situations. We tested the hypotheses that (1) more frequent viewing of television images of the September 11 terrorist attacks was associated with posttraumatic stress disorder (PTSD) and depression, and that (2) direct exposure to disaster events had an interactive effect with media viewing. We recruited 1,008 adult residents of the borough of Manhattan in New York City through a random-digit-dial telephone survey conducted between October 16 and November 15, 2001. Respondents who repeatedly saw "people falling or jumping from the towers of the World Trade Center" had higher prevalence of PTSD (17.4%) and depression (14.7%) than those who did not (6.2% and 5.3%, respectively). Among respondents who were directly affected by the attacks (e.g., had a friend killed), those who watched this television image frequently were more likely to have PTSD and depression than those who did not. Among respondents not directly affected by the attacks, prevalence of PTSD and depression was not associated with frequency of television image viewing. Specific disaster-related television images were associated with PTSD and depression among persons who were directly exposed to a disaster. Future research should address causal directionality of this association. PMID- 12530331 TI - Television viewing as risk factor. PMID- 12530332 TI - Exposure to traumatic images: symptom or cause? PMID- 12530333 TI - Does television viewing satisfy criteria for exposure in posttraumatic stress disorder? PMID- 12530334 TI - Televised trauma and viewer PTSD: implications for prevention. PMID- 12530335 TI - The differential effectiveness of psychosocial and biogenetic causal explanations in reducing negative attitudes toward "mental illness". AB - Given the apparent failure of the "mental illness is an illness like any other" approach to reducing negative stereotypes of people with mental health problems, the differential effects of biogenetic and psychosocial explanations of psychiatric symptoms were evaluated. Attitude measures were administered to young New Zealand adults before and after a video portraying a young man with psychotic symptoms followed by either biogenetic or psychosocial explanations. Consistent with previous studies, the "medical model" approach significantly increased perceptions of dangerousness and unpredictability. Following the psychosocial explanation there was a slight but statistically insignificant improvement in attitudes. Participants who knew users of psychiatric services, or who had used services themselves, had more positive attitudes than other participants. It is recommended that destigmatization programs minimize efforts to persuade the public that mental health problems are biogenetic illnesses, and focus instead on increasing exposure to users of mental health services. Further research is required to determine whether such exposure should include life histories, so as to highlight the psychosocial causes of mental health problems. PMID- 12530336 TI - An assessment of suicide in an urban VA Medical Center. AB - This study characterizes those patients of an urban VA Medical Center (VAMC) who had committed suicide. A cause-of-death search of the 1,075 veterans from the VAMC's case rolls who died during 1998 was conducted. For confirmed and suspected suicides, a chart review was conducted, noting psychiatric history and recent contact with the VAMC. Nineteen patients were either confirmed or suspected suicides. Few of these patients had made recent contact with the VAMC, although the majority of them had received psychiatric services from the VA at some time. The proportion of deaths due to suicide was similar in African American and White patients. Rates of suicide were elevated, primarily because of the high proportion of patients receiving psychiatric treatment. Elderly suicides were less likely to have had psychiatric services or previous psychiatric diagnoses than were younger suicides. Patients with past contact with psychiatric services may be especially at risk of suicide, particularly as contact with these services diminishes. Elderly patients in medical settings with undiagnosed or undertreated psychiatric disorders are also likely to have elevated risk for suicide. These findings demonstrate the importance of acknowledging that risk factors for suicide are specific to sites or populations; this information can be used in allocating resources for developing site-specific strategies for prevention. PMID- 12530337 TI - From passivity to competence: a conceptualization of knowledge, skill, tolerance, and empathy. PMID- 12530338 TI - Social support as a predictor of response to group therapy for complicated grief. AB - This study investigated the effect of perceived social support on the outcome of group therapy for patients who experienced complicated grief following a variety of death losses. One hundred and seven psychiatric outpatients, who received either interpretive or supportive group therapy, rated their perceptions of social support from three sources (family, friends, and a special person) prior to treatment onset. For patients in both forms of therapy, perceived social support from friends was directly associated with favorable treatment outcome. In contrast, perceived social support from family was inversely related to outcome for patients in both forms of therapy. Perceived social support from a special person was directly related to favorable improvement in grief symptomatology for patients in interpretive therapy, but unrelated for those in supportive therapy. The results highlight the importance of assessing the level of support patients perceive from their social networks. The findings also suggest that the effect of perceived social support may also depend on the source of the support. Possible explanations and clinical implications of these findings are discussed. PMID- 12530339 TI - The psychological experience of security officers who work with executions. AB - Open-ended interviews were conducted with members of the "Execution Team" in Louisiana in order to understand the roles, experiences, and effects of carrying out the death penalty. Fifty of a potential 52 correctional officers who work with executions were interviewed and asked to complete mental health inventories. While emphasizing the importance of security and their professional responsibilities in carrying out the death penalty, the officers stress their respect and decency toward the inmates and all others involved with the process. Although expectably reporting high incidences of exposure to trauma and death, they generally are not clinically depressed. They rely upon religious beliefs, group identity, administrative support, and their capacity to dissociate and rely on diffusion of responsibility to suppress painful emotions. Nevertheless, the officers experience conflicted feelings and frequently report having a hard time carrying out society's "ultimate punishment." PMID- 12530340 TI - Risperidone in the treatment of Hispanic inpatients with schizophrenia: a pilot study. AB - A growing body of scientific evidence over the last two decades suggests that certain ethnic groups may require lower dosages of standard antipsychotics for the treatment of schizophrenia symptoms. Recent studies have implicated the role of genetic and environmental factors in the metabolism of these drugs as the basis for this differential response. In this pilot study, 10 Hispanic and 8 non Hispanic patients with schizophrenia were enrolled in a double-blind, parallel group, inpatient risperidone dosing (daily versus twice daily) trial with the novel antipsychotic risperidone. The result of repeated measures ANOVA reveals a significant interaction effect for race, indicative of a faster rate of symptom improvement (PANSS General) in Hispanic patients. The findings suggest that this novel agent may be preferable for certain ethnic groups. A trend toward more frequently occurring extrapyramidal symptoms among Hispanics was also found, which suggests that dosages lower than those typically recommended may be necessary in Hispanic schizophrenics. PMID- 12530341 TI - Advocacy efforts key to progress. Member involvement needed more than ever. PMID- 12530342 TI - Another layer of regulations: research under HIPAA. PMID- 12530343 TI - HIM on the front lines of change. Marching toward the National Health Information Infrastructure. PMID- 12530344 TI - Information infrastructure promises. Better healthcare, lower costs. PMID- 12530345 TI - A shot in the arm for public health. Weak systems require reinforcement at all levels. PMID- 12530346 TI - Managing the transcription revolution. Industry forces shape future of field. AB - You may be struggling with contract issues with a vendor. Or maybe you're contemplating the pros and cons of working with outsource, at-home, or overseas transcriptionists. It's a fact: if transcription processes aren't working efficiently, the entire HIM department may be adversely affected. Factor in additional concerns such as data capture for electronic health records, compliance, and patient safety, and the importance of ensuring quality and cost efficient transcription becomes even more apparent. To help you answer some of these questions, the Journal of AHIMA is launching a four-part series dedicated to transcription issues from the HIM professional's point of view. In this issue, we begin with MTIA president Scott Faulkner's overview of the industry and where it's going next. In upcoming issues, other experts will look at controlling cost and monitoring quality, navigating new technologies, and dealing with contract related issues. PMID- 12530347 TI - Beginnings: "if you do not organize now, you perhaps never will". PMID- 12530348 TI - Preventing risk in long-term care: lessons learned. PMID- 12530349 TI - Practice brief. Defining the designated record set. PMID- 12530350 TI - Moving toward a unified information security program. PMID- 12530351 TI - 2003 OIG Work Plan highlights aberrant coding patterns. Plan puts new spin on 2002 work plans, introduces new projects. PMID- 12530352 TI - Plenty of CPT changes for 2003. Latest on additions, revisions, and deletions. PMID- 12530353 TI - Where to find answers to your coding questions. PMID- 12530354 TI - A review of the status of foot and mouth disease in South-East Asia and approaches to control and eradication. AB - The author presents reports of foot and mouth disease (FMD) submitted between 1996 and 2001 to the Office International des Epizooties (OIE: World organisation for animal health) Sub-Commission for FMD in South-East Asia. Of the ten countries in South-East Asia, FMD is endemic in seven (Cambodia, Laos, Malaysia, Myanmar, the Philippines, Thailand and Vietnam) and three are free of the disease (Brunei, Indonesia and Singapore). Part of the Philippines is also recognised internationally as being free of FMD. From 1996 to 2001, serotype O viruses caused outbreaks in all seven of the endemically infected countries. On the mainland, three different type O lineages have been recorded, namely: the South East Asian (SEA) topotype, the pig-adapted or Cathay topotype and the pan-Asian topotype. Prior to 1999, one group of SEA topotype viruses occurred in the eastern part of the region and another group in the western part. However, in 1999, the pan-Asian lineage was introduced to the region and has become widespread. The Cathay topotype was reported from Vietnam in 1997 and is the only FMD virus currently endemic in the Philippines. Type Asia 1 has never been reported from the Philippines but was reported from all countries on the mainland except Vietnam between 1996 and 2001. Type A virus has not been reported from east of the Mekong River in the past six years and seems to be mainly confined to Thailand with occasional spillover into Malaysia. The distribution and movement of FMD viruses in the region is a reflection of the trade-driven movement of livestock. There is great disparity across the region in the strength and resources of the animal health services and this has a direct impact on FMD control. Regulatory environments are not well developed and enforcement of regulations can be ineffectual. The management of animal movement is quite variable across the region and much market-driven transboundary movement of livestock is unregulated. Formal quarantine approaches are generally not supported by traders or are not available. Vaccination is not used widely as a control tool because of the expense. However, it is applied by the Veterinary Services in Malaysia to control incursions of the disease and there is a mass vaccination programme for large ruminants in Thailand where the Government produces and distributes vaccine. Vaccination is also used by the commercial pig sector, particularly in the Philippines and Thailand. PMID- 12530355 TI - Rapid enlargement of a residual craniopharyngioma during short-term growth hormone replacement. PMID- 12530356 TI - Reactive angioendotheliomatosis and sarcoidosis. PMID- 12530357 TI - Atypical varicella with palm and sole involvement. AB - Varicella is a common disease characterized by a typical presentation. We report a case of an atypical presentation of varicella with a centrifugal distribution, eruption with many vesicles, no pustular stage in evolution and distal involvement. There were none of the known modifying factors (immunosuppression, skin disease, injury or sun exposure). To explain the distal involvement we suggest intraepidermic lesions caused by a pre-existing B1 coxsackie infection. PMID- 12530359 TI - Portable ultrasound scanners. Shrinking size, growing market. PMID- 12530358 TI - Nevus comedonicus with hidradenoma papilliferum and syringocystadenoma papilliferum in the female genital area. PMID- 12530360 TI - GlucoWatch Biographer. An automatic, noninvasive glucose monitor. PMID- 12530361 TI - A group of cranio-cervical acceleration/deceleration trauma patients who developed chronic post-traumatic headache. PMID- 12530363 TI - Gravitational unloading effects on muscle fiber size, phenotype and myonuclear number. AB - The effects of gravitational unloading with or without intact neural activity and/or tension development on myosin heavy chain (MHC) composition, cross sectional area (CSA), number of myonuclei, and myonuclear domain (cytoplasmic volume per myonucleus ratio) in single fibers of both slow and fast muscles of rat hindlimbs are reviewed briefly. The atrophic response to unloading is generally graded as follows: slow extensors > fast extensors > fast flexors. Reduction of CSA is usually greater in the most predominant fiber type of that muscle. The percentage of fibers expressing fast MHC isoforms increases in unloaded slow but not fast muscles. Myonuclear number per mm of fiber length and myonuclear domain is decreased in the fibers of the unloaded predominantly slow soleus muscle, but not in the predominantly fast plantaris. Decreases in myonuclear number and domain, however, are observed in plantaris fibers when tenotomy, denervation, or both are combined with hindlimb unloading. All of these results are consistent with the view that a major factor for fiber atrophy is an inhibition or reduction of loading of the hindlimbs. These data also indicate that predominantly slow muscles are more responsive to unloading than predominantly fast muscles. PMID- 12530364 TI - The morbidity of Guillain-Barre syndrome admitted to the intensive care unit. AB - Patients with severe forms of Guillain-Barre syndrome (GBS) require intensive care. Specific treatment, catheterization, and devices may increase morbidity in the intensive care unit (ICU). To understand the spectrum of morbidity associated with ICU care, the authors studied 114 patients with GBS. Major morbidity occurred in 60% of patients. Complications were uncommon if ICU stay was less than 3 weeks. Respiratory complications such as pneumonia and tracheobronchitis occurred in half of the patients and were linked to mechanical ventilation. Systemic infection occurred in one-fifth of patients and was more frequent with increasing duration of ICU admission. Direct complications of treatment and invasive procedures occurred infrequently. Life-threatening complications such as gastrointestinal bleeding and pulmonary embolism were very uncommon. Pulmonary morbidity predominates in patients with severe GBS admitted to the ICU. Attention to management of mechanical ventilation and weaning is important to minimize this complication of GBS. Other causes of morbidity in a tertiary center ICU are uncommon. PMID- 12530365 TI - Changes for long term care Medicare/Medicaid certification-based option. PMID- 12530366 TI - Hypergravity modulates behavioral nociceptive responses in rats. AB - Hypergravity (2G) exposure elevated the nociceptive threshold (pain suppression) concomitantly with evoked neuronal activity in the hypothalamus. Young Wistar male rats were exposed to 2G by centrifugal rotation for 10 min. Before and after 2G exposure, the nociceptive threshold was measured as the withdrawal reflex by using the von Frey type needle at a total of 8 sites of each rat (nose, four quarters, upper and lower back, tail), and then rats were sacrificed. Fos expression was examined immunohistochemically in the hypothalamic slices of the 2G-treated rats. When rats were exposed to 2G hypergravity, the nociceptive threshold was significantly elevated to approximately 150 to 250% of the 1G baseline control levels in all the examination sites. The 2G hypergravity remarkably induced Fos expression in the paraventricular and arcuate nuclei of the hypothalamus. The analgesic effects of 2G hypergravity were attenuated by naloxone pretreatment. Data indicate that hypergravity induces analgesic effects in rats, mediated through hypothalamic neuronal activity in the endogenous opioid system and hypothalamo-pituitary-adrenal axis. PMID- 12530367 TI - Life support for aquatic species--past; present; future. AB - Life Support is a basic issue since manned space flight began. Not only to support astronauts and cosmonauts with the essential things to live, however, also animals which were carried for research to space etc. together with men need support systems to survive under space conditions. Most of the animals transported to space participate at the life support system of the spacecraft. However, aquatic species live in water as environment and thus need special developments. Research with aquatic animals has a long tradition in manned space flight resulting in numerous life support systems for them starting with simple plastic bags up to complex support hardware. Most of the recent developments have to be identified as part of a technological oriented system and can be described as small technospheres. As the importance arose to study our Earth as the extraordinary Biosphere we live in, the modeling of small ecosystems began as part of ecophysiological research. In parallel the investigations of Bioregenerative Life Support Systems were launched and identified as necessity for long-term space missions or traveling to Moon and Mars and beyond. This paper focus on previous developments of Life Support Systems for aquatic animals and will show future potential developments towards Bioregenerative Life Support which additionally strongly benefits to our Earth's basic understanding. PMID- 12530368 TI - A collection of studies on taste disturbance from the Department of Otorhinolaryngology, Nihon University School of Medicine. Memorial issue to Emeritus Professor Hiroshi Tomita, at his retirement as the chairman of the department. PMID- 12530369 TI - On the Born-Alive Infants Protection Act of 2000. PMID- 12530370 TI - Cloning patent raises important ethical questions. PMID- 12530371 TI - The potentiality argument in the debate relating to the beginning of personhood. PMID- 12530372 TI - Prenatal diagnosis and ethics: a Turkish evaluation. PMID- 12530373 TI - X-Y sperm selection: fact or fiction? AB - Selecting the gender of offspring has given rise to various and sometimes amusing stories. But regardless of which prefertilisation technique is used to influence the sex ratio of offspring it must fulfill certain criteria. First of all it must achieve a complete separation of the X and Y bearing sperm in sufficient quantities. Secondly sperm must be viable after separation and capable of fertilising. Sex preselection methods can be divided into two general groups which either separate spermatozoa on the basis of subtle physical or kinetic features or those which rely on distinctive nuclear characteristics unique either to X or Y chromosome bearing sperm. These, in turn, can be divided into in vivo methods designed to produce optimal conditions for fertilisation by either the X or Y bearing sperm, or in vitro sperm separation methods designed to separate X or Y bearing sperm. According to all published data, the different separation techniques have been shown not to be very effective. Only sex selection of spermatozoa by chromatin differences (cell sorting by flow cytometry) has demonstrated a significant enrichment of the X bearing sperm. PMID- 12530374 TI - Legislating for the new predictive genetics. AB - Many new genetic markers have become available for use in the diagnosis, prognosis or risk prediction of common multifactorial disease such as venous thrombo-embolism, coronary artery disease, dementias and some cancers. Regulation or legislation of their application in the fields of the family, employment, life assurance, confidentiality and property law is required. This is made difficult because of the rapid pace of genetic discoveries and their derived technologies, the diversity of opinions on the legitimate application of these new techniques, and the pluralistic and evolving social norms of society regarding the use of the new genetic methods. This paper examines some of the problems that can arise when regulation is attempted in each of the above fields. A variety of solutions such as referenda, moratoria, ethical codes of professional bodies or the drafting of scientifically accurate and appropriate legislation depending on particular circumstances are discussed as a means of achieving a flexible and responsive approach to the challenges posed by the use of the new genetic markers. PMID- 12530375 TI - NASDA aquatic animal experiment facilities for Space Shuttle and ISS. AB - National Space Development Agency of Japan (NASDA) has developed aquatic animal experiment facilities for NASA Space Shuttle use. Vestibular Function Experiment Unit (VFEU) was firstly designed and developed for physiological research using carp in Spacelab-J (SL-J, STS-47) mission. It was modified as Aquatic Animal Experiment Unit (AAEU) to accommodate small aquatic animals, such as medaka and newt, for second International Microgravity Laboratory (IML-2, STS-65) mission. Then, VFEU was improved to accommodate marine fish and to perform neurobiological experiment for Neurolab (STS-90) and STS-95 missions. We have also developed and used water purification system which was adapted to each facility. Based on these experiences of Space Shuttle missions, we are studying to develop advanced aquatic animal experiment facility for both Space Shuttle and International Space Station (ISS). PMID- 12530377 TI - Aquatic animal research in space station and its issues--focus on support technology on nitrate toxicity. AB - We studied the effects of accumulated nitrate in water on the spawning, hatching and development of medaka using a simple nitrifying filter and a combined filter having both nitrifying and denitrifying capabilities. A nitrate concentration of 100 mg NO3(-)-N/L was clearly of lethal toxicity to fish when they were exposed to nitrate in both adult and the growing phases. A nitrate concentration of 75 mg NO3(-)-N/L reduced the fertilization rate, delayed the hatching time and reduced the hatching rate of the eggs laid by adults and decreased the growth rate of juveniles. In addition, nitrate accumulations as low as 50 mg NO3(-)-N/L remarkably retarded spawning and lowered the number of eggs laid by fish exposed in the juvenile phase. The effects on the reproduction system may be initiated by a low concentration, approximately 30 mg NO3(-)-N/L. PMID- 12530376 TI - Mutagenic target for hydroxyl radicals generated in Escherichia coli mutant deficient in Mn- and Fe- superoxide dismutases and Fur, a repressor for iron uptake systems. AB - We previously reported that mutations in Mn- and Fe-superoxide dismutases and Fur, a repressor for iron uptake systems, simulated generation of hydroxyl radicals, and caused hypermutability in Escherichia coli. The predominant type of spontaneous mutation was GC --> TA, followed by AT --> CG, suggesting the involvement of 7,8-dihydro-8-oxoguanine (8-oxoG) and 1,2-dihydro-2-oxoadenine (2 oxoA) in DNA as well as 7,8-dihydro-8-oxodeoxyguanosine triphosphate (8-oxodGTP) and 1,2-dihydro-2-oxodeoxyadenosine triphosphate (2-oxodATP) in the nucleotide pool. To determine the targets contributing to oxidative mutagenesis, DNA or nucleotides, we characterized spontaneous mutations and compared the distribution to those in mutMY and mutT strains, in which GC --> TA and AT --> CG were predominantly induced, respectively. The hotspots and sequence contexts where AT -> CG occurred frequently in sodAB fur strain were almost identical to those in mutT strain,whereas, those where GC --> TA occurred frequently in sodAB fur strain were quite different from those in mutMY strain. These observations suggested that AT --> CG is due to 8-oxodGTP, while GC --> TA is produced by some other lesion(s). The 2-oxodATP is also a major oxidative lesion in nucleotides, and strongly induces GC --> TA. The expression of cDNA for MTH1, which can hydrolyze 2-oxodATP as well as 8-oxodGTP, partially but significantly, suppressed the GC --> TA mutator phenotype of the sodAB fur strain, whereas, it did not for the mutMY strain. Additionally, the sequence contextby 2-oxodATP in E. coli was similar to that in sodAB fur strain. These results suggested that the targets contributing to oxidative mutagenesis in sodAB fur strain are nucleotides such as dGTP and dATP, rather than DNA. PMID- 12530378 TI - Recommendations for federal "super agency" would affect institutional review boards. PMID- 12530379 TI - New privacy rule has more tasks for IRBs. PMID- 12530380 TI - Accreditation standards are proposed for IRBs. PMID- 12530381 TI - Institutional review board (IRB) violated numerous regulations. PMID- 12530382 TI - Researcher sues government and specific agency official over misconduct investigation. PMID- 12530383 TI - Experiments with small animals in BIOLAB and EMCS on the International Space Station. AB - Two ESA facilities will be available for animal research and other biological experiments on the International Space Station: the European Modular Cultivation System (EMCS) in the US Lab "Destiny" and BIOLAB in the European "Columbus" Laboratory. Both facilities use standard Experiment Containers, mounted on two centrifuge rotors allowing either research in microgravity or acceleration studies with variable g-levels from 0.001 to 2.0 x g. Standard interface plates provide each container with power and data lines, gas supply (controlled CO2, O2 concentration and relative humidity), and--for EMCS only--connectors to fresh and waste water reservoirs. The experiment hardware inside the containers will be developed by the user, but ESA conducted a feasibility study for several kinds of Experiment Support Equipment with potential use for research on small animals: design concepts for experiments with insects, with aquatic organisms like rotifers and nematodes, and with small aquatic animals (sea urchin larvae, tadpoles, fish youngsters) are described in detail in this presentation. Also ESA's initial steps to support experiments with rodents on the Space Station are presented. PMID- 12530384 TI - The future of mothering: reproductive technology and feminist theory. AB - An exploration of (I) alternative perspectives toward recent innovations in reproductive technology: support for new techniques for the sake of the kind of feminist future they facilitate; unqualified opposition despite therapeutic benefit to individual women; or qualified opposition depending upon specific threats to women's interests and (II) relationships between these positions and values bound up with mothering practices. PMID- 12530385 TI - Comprehensive performance management in the operating room. AB - A clearly defined method for measuring and analyzing operating room (OR) processes provides a common language for focusing management's attention on areas of variability that can be eliminated. But hospitals must also assess OR performance against multiple financial aid and clinical quality metrics to achieve continuous improvement in patient outcomes and profitability enterprisewide. This project is a collaborative effort by McKesson Information Solutions and the Healthcare Financial Management Association. PMID- 12530386 TI - The ethics of biotechnology. AB - Ethical considerations pertaining to biotechnology have progressively been introduced into European community law, especially with regard to research, the environment, patents, animal welfare and medicine. This article addresses the ethics of biotechnology at the European Union (EU) level. It provides insight into the European Group of Ethics and describes how European ethical values concerning science and technology are defined by law and intimately connected to Europe's cultural and political identity. In addition, this article highlights the critical need for an independent and permanent international body to spearhead discussion and collaboration so that views between the different cultures of the world can be exchanged, and, to assure access to experts whose legitimacy and accountability are universally recognized. PMID- 12530387 TI - [30th Annual Meeting of the Austrian Society of Diabetes. 28-30 November 2002, Graz. Abstracts]. PMID- 12530388 TI - Crickets in space: morphological, physiological and behavioral alterations induced by space flight and hypergravity. AB - "Crickets in Space" was a Neurolab experiment by which the balance between genetic programs and the gravitational environment for the development of a gravity sensitive neuronal system was studied. The model character of crickets was justified by their external gravity receptors, identified position-sensitive interneurons (PSI) and gravity-related compensatory head response, and by the specific relation of this behavior to neuronal arousal systems activated by locomotion. These advantages allowed to study the impact of modified gravity on cellular processes in a complex organism. Eggs, 1st, 4th and 6th stage larvae of Acheta domesticus were used. Post-flight experiments revealed a low susceptibility of the behavior to micro- and hypergravity while the physiology of the PSI was significantly affected. Immunocytological investigations revealed a stage-dependent sensitivity of thoracic GABAergic motoneurons to 3 g-conditions concerning their soma sizes but not their topographical arrangement. The morphology of neuromuscular junctions was not affected by 3 g-hypergravity. Peptidergic neurons from cerebral sensorimotor centers revealed no significant modifications by microgravity (micro g). The contrary physiological and behavioral results indicate a facilitation of 1 g-readaptation originating from accessory gravity, proprioceptive and visual sense organs. Absence of anatomical modifications point to an effective time window of micro g or 3 g-expo-sure related to the period of neuronal proliferation. The analysis of basic mechanisms of how animals and man adapt to altered gravitational conditions will profit from a continuation of the project "Crickets in Space". PMID- 12530389 TI - Biosamples, genomics, and human rights: context and content of Iceland's Biobanks Act. AB - In recent years, human DNA sampling and collection has accelerated without the development of enforceable rules protecting the human rights of donors. The need for regulation of biobanking is especially acute in Iceland, whose parliament has granted a for-profit corporation, deCODE Genetics, an exclusive license to create a centralized database of health records for studies on human genetic variation. Until recently, how deCODE Genetics would get genetic material for its genotypic phenotypic database remained unclear. However, in May 2000, the Icelandic Parliament passed the Icelandic Biobanks Act, the world's earliest attempt to construct binding rules for the use of biobanks in scientific research. Unfortunately, Iceland has lost an opportunity for bringing clear and ethically sound standards to the use of human biological samples in deCODE's database and in other projects: the Biobanks Act has extended a notion of "presumed consent" from the use of medical records to the use of patients' biological samples; worse, the act has made it possible--perhaps likely--that a donor's wish to withdraw his/her sample will be ignored. Inadequacies in the Act's legislative process help account for these deficiencies in the protection of donor autonomy. PMID- 12530390 TI - International Association for Dental Research divisional abstracts, 2001, 2002. PMID- 12530391 TI - Straight talk: the dreaded. PMID- 12530392 TI - Yale School of Medicine thesis abstracts 2002. PMID- 12530393 TI - 2003 Gold Book Buyer's Guide. PMID- 12530394 TI - Emergency Medical Services 25th annual state & province survey. PMID- 12530395 TI - Directory of government agencies. PMID- 12530396 TI - Directory of EMS organizations. PMID- 12530397 TI - Neuronal feedback between brain and inner ear for growth of otoliths in fish. AB - Previous investigations revealed that fish inner ear otolith growth (concerning otolith size and calcium-incorporation) depends on the amplitude and the direction of gravity, suggesting the existence of a (negative) feedback mechanism. In search for the regulating unit, the vestibular nerve was unilaterally transected in neonate swordtail fish (Xiphophorus helleri) which were subsequently incubated in the calcium-tracer alizarin-complexone. Calcium incorporation ceased on the transected head sides, indicating that calcium uptake is neurally regulated. PMID- 12530398 TI - Protein kinase C regulation of rat jejunal transport system: mechanisms involved in lactate movement. AB - We examined whether protein kinase C (PKC) modulates the transport systems involved in lactate movements across the plasma membranes of rat jejunum. In vitro phosphorylated membrane vesicles were used to perform uptake studies, the results of which suggested that PKC activation exerts an inhibitory effect on basolateral H+-lactate symport, as well as on apical N-+glucose cotransport. The specificity of the response to PKC was confirmed by using staurosporine, chelerythrine or 4-alpha-PMA. Experiments performed using the whole tissue incubated in vitro confirmed the reduction of lactate transport elicited by PKC and gave evidence for an associated inhibition of fluid transport. Na+K+-ATPase activity seems to be unaffected by the kinase and inhibited by Ca2+. Taken together, our results suggest that the overall action of PKC results from the simultaneous modulation of multiple pathways, targeted to a reduction of both lactate and bicarbonate transports without altering cell pH homeostasis. PMID- 12530399 TI - Neuropeptides, amines and amino acids as mediators of the sympathetic effects of paraventricular nucleus activation in the rat. AB - The aim of the present study was to determine the influence on renal sympathetic nerve activity of the different chemically coded neuronal phenotypes that project from the paraventricular nucleus (PVN) to the spinal cord. Experiments were carried out on male Wistar rats anaesthetised with chloralose and urethane. Changes in renal sympathetic nerve activity were measured following activation of neurones in the PVN with D,L-homocysteic acid (100 nl, 200 mM), before and following intrathecal application of glutamate, vasopressin, oxytocin, dopamine and their receptor antagonists. Excitatory and inhibitory effects on renal sympathetic nerve activity were elicited by PVN stimulation. PVN excitatory effects were mimicked by intrathecal administration of glutamate and vasopressin and selectively antagonised by intrathecal administration of kynurenic acid and a V1a receptor antagonist, respectively. A low dose of dopamine increased renal sympathetic activity and this was selectively antagonised by haloperidol; however, the latter was without effect on PVN excitatory responses. A high dose of dopamine decreased renal sympathetic nerve activity and this was selectively blocked by a D1 dopamine receptor antagonist (SCH 23390), which also antagonised a minority of inhibitory responses obtained from the caudal extension of the PVN. Oxytocin also had two actions in 5 rats it inhibited and in 10 rats it increased renal sympathetic nerve activity, both actions being blocked selectively by oxytocin receptor antagonists. Neither of the PVN effects on renal sympathetic nerve activity appeared to be dependent on oxytocin pathways. Tests with intrathecal administration of bicuculline showed that PVN inhibition of renal sympathetic nerve activity was not dependent on spinal GABA(A) receptor activation. The results show that PVH-induced excitation of sympathetic activity to the kidney is mainly mediated by glutamate or vasopressin neurones whereas dopamine via Dl receptors may mediate some of the PVN inhibitory effects. PMID- 12530400 TI - Do cardiac neurons play a role in the intrinsic control of heart rate in the rat? AB - Three experiments were performed to see whether cardiac neurons contribute to the intrinsic control of heart rate in right atria of adult rats. The intrinsic heart rate response (IRR) was examined by raising right atrial pressure from 2 to 8 mmHg for 3 min. In isolated preparations of the right atrium, the IRR was not significantly altered by the addition of either 1 microM atropine (n =6; control +19+/- 3 min(-1) ; atropine+18+/-3 min(-1); (mean /+/-S.E.M.)) or 1 microM propranolol (n = 5; control +22+/- 4 min(-1); ; propranolol +21+/-3 min(-1); ). Tetrodotoxin (0.5 microm) had no effect on the IRR (n = 6; control +37+/-5 min( 1); tetrodotoxin 38+/-5 min(-1); ). In another experiment, 2-day-old rat pups were injected with capsaicin (50 mg kg(-1); treated) or with vehicle(control). There was no difference in the IRR of right atrial preparations taken from control and treated animals after they reached adulthood (control (n = 7) and treated (n = 8): +30+/- 4 and +32+/- 4 min(-1)). The influence of right atrial pressure on the efficacy of vagal stimulation was examined. The rate response to vagal stimulation was reduced similarly in control and treated preparations when pressure was elevated from 2 to 4 mmHg (control and treated: -34+/- 5% and -33+/- 3%). The effectiveness of the capsaicin treatment was confirmed by the depletion of substance P-immunoreactive nerve fibres in cardiac tissues. Together, these results strongly suggest that cardiac neurons are not involved in intrinsic heart rate control. PMID- 12530401 TI - Remote noxious stimuli modulate jaw reflexes evoked by activation of periodontal ligament mechanoreceptors in man. AB - The purpose of the study was to investigate whether jaw reflexes evoked by selective stimulation of periodontal ligament me canoreceptors are susceptible to modulation by remote noxious stimulation. Experiments were performed on 10 volunteer subjects. Skin surface recordings were made from the jaw-closing masseter muscle. The subjects activated the muscle to approximately 10% of maximum by biting on a rubber impression of their molar teeth while they received visual feedback of the electromyogram (EMG) of the muscle. Reflexes were produced by the application of gentle mechanical stimuli to an upper central incisor tooth. The stimuli were in the form of 'ramp and hold' forces with a 5 ms rise time and a 1.5 N plateau which lasted 350 ma. The resulting reflexes were recorded both under control conditions and while the subjects received a remote noxious stimulus (immersion of a hand in water at 3 degrees C). In all 10 subjects, the stimuli produced a single period of inhibition of masseteric activity (latency, 12.8 t 04 ms; duration, 18.1+/-1.3 ms; means +/- S.E.M.), which was usually followed by a period of increased masseteric activity. The period of inhibition constituted a downward wave in full-wave rectified, averaged signals. The integrals of such waves were significantly smaller (by 17+/- 6.5 %; P = 0.027; Student's t test) when the reflex was evoked during remote noxious stimulation rather than under control conditions. As such reflexes are beLieved to play a modulatory role during normal oral function this finding maybe relevant to disorders of mastication associated with pain. PMID- 12530402 TI - Subthalamic DBS replaces levodopa in Parkinson's disease: two-year follow-up. PMID- 12530403 TI - Advantages and risks of multiphoton microscopy in physiology. AB - Multiphoton microscopy is based on the simultaneous absorption of two photons emitted by a pulsed infrared laser source. In this technique, the excitation is restricted to a very small focus and thus results in optical sectioning a priori without the need of a confocal aperture. Multiphoton microscopy was introduced in live cell imaging as an alternative to confocal microscopy due to its superior qualities, such as the deep penetration depth, the reduced photodamage and the lack of out-of-focus bleaching. However, during the past years, examinations revealed severe limitations to the initial expectations. In the focal plane, photodamage and photobleaching can be worse than in single photon microscopy. However, studies showed that with low excitation intensity and by special technical adaptations photodamage could be avoided successfully. For functional biological imaging multiphoton excitation provides an excellent tool such as the release of caged compounds in a diffraction-limited volume combined with multiphoton or confocal imaging. PMID- 12530404 TI - Calcium dynamics in spines: link to synaptic plasticity. AB - Dendritic spines are morphologically and functionally heterogeneous. Here, we use two-photon imaging of layer V pyramidal neurons in slices from mouse visual cortex to characterize differences in spine calcium dynamics between individual spines. By measuring action potential-evoked [Cell transients in spines, we find different calcium dynamics in spines from proximal apical and distal apical dendrites. Using a mathematical multi-compartmental model, we demonstrate that these differences are even more pronounced in the absence of exogenous calcium buffers. We also find that these different calcium dynamics cause different susceptibility to synaptic depression in proximal and distal apical synapses, and that modifying calcium decay kinetics in spines changes the expression of long term depression. We conclude that the location of the spine determines its time window of calcium compartmentalization and degree of calcium-dependent synaptic plasty. Our results highlight the precision of the design of neocortical neurons. PMID- 12530405 TI - Combined confocal microscopy and stereology: a highly efficient and unbiased approach to quantitative structural measurement in tissues. AB - Understanding the relationship of the structure of organs to their function is a key component of integrative physiological research. The structure of the organs of the body is not constant but changes, both during growth and development and under conditions of sustained stress (e g. high altitude exposure and disease). Recently, powerful new techniques have become available in molecular biology, which promise to provide novel insights into the mechanisms and consequences of these altered structure-function relationships. Conventionally structure-function relationships are studied by microscopic examination of tissue sections. However, drawing conclusions about the three-dimensional structure of an organ based on this two-dimensional information frequently leads to serious errors. The techniques of stereology allow precise and accurate quantification of structural features within three-dimensional organs that relate in a meaningful way to integrated function. For example, knowledge of changes in the total surface area of the capillary endothelium in a n organ can be related directly to changes in fluid filtration and permeability, or knowledge of total vessel length and mean radius allows deductions about vascular resistance. Confocal microscopy add s enormously to the power of stereological approaches. It reduces the difficulties and labour involved in obtaining suitable images. Moreover, when used in conjunction with new analytical software, it allows convenient application of stereology to small samples and those in which it is essential to maintain a specific orientation for interpretation. The information obtained will allow us to examine in a quantitative manner the altered structure-function relationships produced by manipulation of single genes and regulatory pathways in whole organisms. PMID- 12530408 TI - Abstracts of the 4th World Congress on Stress. Edinburgh, United Kingdom, September 12-15, 2002. PMID- 12530406 TI - Retinal arteriolar narrowing and risk of coronary heart disease. PMID- 12530407 TI - Abstracts of the Society of Surgical Oncology 56th Annual Cancer Symposium. Los Angeles, California, USA. March 5-9, 2003. PMID- 12530409 TI - Abstracts of the Xth International Congress on Neuromuscular Diseases. 7-12 July 2002, Vancouver, Canada. PMID- 12530410 TI - Heme oxygenase-1 and mice in red, white, and blue. PMID- 12530411 TI - Invited comment on W. Ebstein: On the therapy of diabetes mellitus, in particular on the application of sodium salicylate. PMID- 12530412 TI - Effects of altered gravity on the swimming behaviour of fish. AB - Humans taking part in parabolic aircraft flights (PAFs) may suffer from space motion sickness-phenomena (SMS, a kinetosis). It has been argued that SMS during PAFs might not be based on microgravity alone but rather on changing accelerations from 0 g to 2 g. We test here the hypothesis that PAF-induced kinetosis is based on asymmetric statoliths (i.e., differently weighed statoliths on the right and the left side of the head), with asymmetric inputs to the brain being disclosed at microgravity. Since fish frequently reveal kinetotic behaviour during PAFs (especially so-called spinning movements and looping responses), we investigated (1) whether or not kinetotically swimming fish at microgravity would have a pronounced inner ear otolith asymmetry and (2) whether or not slow translational and continuously changing linear (vertical) acceleration on ground induced kinetosis. These latter accelerations were applied using a specially developed parabel-animal-container (PAC) to stimulate the cupular organs. The results suggest that the fish tested on ground can counter changing accelerations successfully without revealing kinetotic swimming patterns. Kinetosis could only be induced by PAFs. This finding suggests that it is indeed microgravity rather than changing accelerations, which induces kinetosis. Moreover, we demonstrate that fish swimming kinetotically during PAFs correlates with a higher otolith asymmetry in comparison to normally behaving animals in PAFs. PMID- 12530413 TI - Restless legs syndrome induced by mirtazapine. PMID- 12530414 TI - Behavior therapy attenuates clozapine-induced obsessions and compulsions. PMID- 12530415 TI - Olanzapine treatment for patients with schizophrenia and cocaine abuse. PMID- 12530416 TI - Interactions in the microbial world. Proceedings of the 9th International Symposium on Microbial Ecology. PMID- 12530417 TI - Limbic leucotomy in self-mutilation. PMID- 12530418 TI - Fluvoxamine treatment of a Japanese patient with koro. PMID- 12530419 TI - Sildenafil advertising and the realities of sildenafil treatment. PMID- 12530420 TI - Hypnotic medication in the aftermath of trauma. PMID- 12530421 TI - Microgravity (STS-90 Neurolab-Mission) influences synapse formation in a vestibular nucleus of fish brain. AB - Synapse counting was undertaken by conventional electron microscopy in primary vestibular integration centers (i.e., Nucleus descendens, Nd, and Nucleus magnocellularis, Nm, of the brainstem Area octavolateralis) and in the diencephalic visual Nucleus corticalis (Nc) of spaceflown neonate swordtail fish Xiphophorus helleri as well as in 1 g control siblings. Spaceflight (16 days microgravity, STS-90 Neurolab-Mission) yielded an increase in synaptic contacts only within the vestibular Nd indicating that lack of input resulted in compensation processes. No effect of microgravity, however, was observed in the visual Nc and in the vestibular Nm which is situated in the close vicinity of the Nd. In contrast to the latter, the Nm does not receive exclusively vestibular input, but inputs from the lateral line as well, possibly providing sufficient input at microgravity. PMID- 12530422 TI - A case report of quetiapine-related tic-like symptoms. PMID- 12530423 TI - A case of ziprasidone-induced mania and the role of 5-HT2A in mood changes induced by atypical antipsychotics. PMID- 12530425 TI - Kinetics of insulin release in health and type 2 diabetes. Proceedings of the 2nd Servier-IGIS Symposium. St. Jean Cap Ferrat, France, 22-24 March 2001. PMID- 12530424 TI - Proceedings of the 11th International Leksell Gamma Knife Meeting. Prague, Czech Republic, 12-16 May 2002. PMID- 12530426 TI - Adherence to HIV drug therapy. AB - Antiretroviral therapy has dramatically improved the prognosis for many patients with HIV infection. For many patients who can navigate combinations of drugs and time their doses precisely, these drug regimens typically slow the progression of the disease and lengthen survival. But because these drug regimens are very complex, adherence--the degree to which patients follow medical advice in taking the prescribed drugs--is now a major determinant of HIV treatment success. This Issue Brief summarizes recent work on the effect of adherence on short-term outcomes, and the ability of providers to predict and estimate their patients' adherence to therapy. PMID- 12530427 TI - Central mechanisms of cardiovascular control--cellular, molecular and integrative aspects. Proceedings of a satellite meeting of the XXXIV Congress of the International Union of Physiological Sciences. Christchurch, New Zealand, 26-30 August 2001. PMID- 12530429 TI - [Is it all over with X-ray small intestine imaging?]. PMID- 12530428 TI - [Prospective feasibility study of chest X-ray vs. thoracic MRI in breath-hold technique at an open low-field scanner]. PMID- 12530430 TI - Effects of hypergravity on the development of cell number and asymmetry in fish brain nuclei. AB - Larval cichlid fish (Oreochromis mossambicus) siblings were subjected to 3 g hypergravity (hg) and total darkness for 21 days during development and subsequently processed for conventional histology. Further siblings reared at 1 g and alternating light/dark (12h:12h) conditions served as controls. Cell number counts of the visual Nucleus isthmi (Ni) versus the vestibular Nucleus magnocellularis (Nm) revealed that in experimental animals total cell number was decreased in the Ni, possibly due to retarded growth as a result of the lack of visual input whereas no effect was observed in the Nm. Calculating the percentual asymmetry in cell number (i.e., right vs. the left side of the brain), no effects of hg/darkness were seen in the Ni, whereas asymmetry was slightly increased in the Nm. Since the asymmetry of inner ear otoliths is decreased under hg, this finding may indicate efferent vestibular action of the CNS on the level of the Nm by means of a feedback mechanism. PMID- 12530431 TI - DNA fragmentation by charged particle tracks. AB - High-LET (linear energy transfer) charged particles induce DNA double-strand breaks (DSB) in a non-random fashion in mammalian cells. The clustering of DSB, probably determined by track structure as well as chromatin conformation, results in an excess of small- and intermediate-sized DNA fragments. DNA fragmentation in normal human fibroblasts (GM5758) was analyzed by pulsed-field gel electrophoresis after irradiation with photons (60Co) or 125 keV/micrometers nitrogen ions. Compared to conventional DSB analysis, i.e. assays only measuring the fraction of DNA smaller than a single threshold, the relative biological effectiveness (RBE) for DSB induction increased with 100%. Further, the size distribution of DNA fragments showed a significant dependence on radiation quality, with an excess of fragments up to 1 Mbp. Irradiation of naked genomic DNA without histone proteins increased the DSB yields 25 and 13 times for photons and nitrogen ions, respectively. The results suggest possible roles of both track structure and chromatin organization in the distribution of DNA double-strand breaks along the chromosome. PMID- 12530432 TI - The conflict between ethics and business in community pharmacy: what about patient counseling? AB - Patient counseling is a cornerstone of ethical pharmacy practice and high quality pharmaceutical care. Counseling promotes patient compliance with prescription regimens and prevents dangerous drug interactions and medication errors. Counseling also promotes informed consent and protects pharmacists against legal risks. However, economic, social, and technological changes in pharmacy practice often force community pharmacists to choose between their professional obligations to counsel patients and business objectives. State and federal legislatures have enacted laws that require pharmacists to counsel patients, but these laws have had mixed results. This essay argues that community pharmacy's patient counseling conundrum can be solved through additional moral education persuasion, not through additional legal mandates. PMID- 12530433 TI - Radiation quality and tissue-specific microenvironments following exposure to 1 GeV/amu Fe. AB - This paper summarizes quantitative in vivo laminin immunofluorescence analysis of mammary glands and skin epithelial structures from mice exposed to 1 GeV/amu Fe ions. Digital confocal microscopic images were quantified and linked to the rough "core-penumbra" Fe track physical description. Comparison to gamma-ray sparsely ionizing radiation suggested the core of the Fe track being responsible for a biological response only seen with energetic Fe particles. Conclusions for modeling in vivo responses to radiation were then implied. PMID- 12530434 TI - Warfarin, aspirin, or both after myocardial infarction. PMID- 12530435 TI - Warfarin, aspirin, or both after myocardial infarction. PMID- 12530436 TI - Ethics counselors as a new priesthood. AB - The aim of this paper is to critically evaluate the thesis that ethics counselors constitute a new priesthood in the pejorative sense of this term. In defense of the thesis, an account is given of the diverse variety of fundamental ideas about ethics or morality. The underlying argument is simply that there is such a diversity of opinion about so many fundamental issues that most ethical appraisals, especially in committees, are probably very shallow and barely warranted. Following this negative work, an attempt is made to try to find some positive benefits from the work of ethics counselors. Some potential benefits are identified, but there is a need for empirical research in order to construct a more persuasive case for such work. In the penultimate section of the paper I addressed some of my own second thoughts about the discussion and some provocative suggestions that friends gave me about earlier drafts. PMID- 12530437 TI - Investigating the cellular effects of isolated radiation tracks using microbeam techniques. AB - Studies of the effects of radiation at the cellular level have generally been carried out by exposing cells randomly to the charged-particle tracks of a radiation beam. Recently, a number of laboratories have developed techniques for microbeam irradiation of individual cells. These approaches are designed to remove much of the randomness of conventional methods and allow the nature of the targets and pathways involved in a range of radiation effects to be studied with greater selectivity. Another advantage is that the responses of individual cells can be followed in a time-lapse fashion and, for example, processes such as "bystander" effects can be studied clearly. The microbeam approach is of particular importance in mechanistic studies related to the risks associated with exposure to low fluences of charged particles. This is because it is now possible to determine the actions of strictly single particle tracks and thereby mimic, under in vitro conditions, exposures at low radiation dose that are significant for protection levels, especially those involving medium- to high-LET radiations. Overall, microbeam methods provide a new dimension in exploring mechanisms of radiation effect at the cellular level. Microbeam methods and their application to the study of the cellular effects of single charged-particle traversals are described. PMID- 12530438 TI - Botulinum toxin for spasticity after stroke. PMID- 12530440 TI - Abstracts of the 5th European Conference on Systemic Lupus Erythematosus. 26-30 May 2002, Athens, Greece. PMID- 12530439 TI - Botulinum toxin for spasticity after stroke. PMID- 12530441 TI - Healthcare ethics: a patient-centered decision model. AB - A common financial model used in business decisions is the cost/benefit comparison. The costs of a proposed project are compared with the benefits, and if the benefits outweigh the costs, the project is accepted; if the costs exceed the benefits, the project is rejected. This model is applicable when tangible costs and benefits can be reasonably measured in monetary units. However, it is difficult to consider intangible factors in this model because intangible factors cannot be readily quantified in money. While some might argue that the financial model should not apply to healthcare decisions, the fact is that costs do enter into the picture. People may decide to forego needed healthcare because they cannot afford it. Healthcare providers may make choices based in part on the costs of diagnosis and treatment, rather than solely on medical information and what is best for the patient. Should financial issues enter into healthcare decisions--decisions about human health and well being? If so, how should the costs and benefits be measured and evaluated? What are some ethical issues and dilemmas involved in such decisions. This paper addresses ethical dilemmas and financial issues in healthcare. A healthcare decision model, which considers medical information, financial information, as well as ethical and other intangible factors, is proposed. PMID- 12530442 TI - Health care law--Medicaid--Third Circuit finds providers lack standing to enforce the Medicaid Act.--Pennsylvania Pharmacists Ass'n v. Houstoun, 283 F.3d 531 (3d Cir. 2002), cert. denied, 123 S.Ct. 100 (2002). PMID- 12530443 TI - Increase in nocturnal blood pressure and progression to microalbuminuria in diabetes. PMID- 12530444 TI - Increase in nocturnal blood pressure and progression to microalbuminuria in diabetes. PMID- 12530445 TI - More on the use of near-infrared spectroscopy to measure muscle oxygenation in humans. PMID- 12530446 TI - Increase in nocturnal blood pressure and progression to microalbuminuria in diabetes. PMID- 12530447 TI - Integrating research ethics and undercover hospital fieldwork. PMID- 12530448 TI - Simultaneous exposure of mammalian cells to heavy ions and X-rays. AB - Crews of space missions are exposed to a mixed radiation field, including sparsely and densely ionizing radiation. To determine the biological effectiveness of mixed high-/low-LET radiation fields, mammalian cells were exposed in vitro simultaneously to X-rays and heavy ions, accelerated at the HIMAC accelerator. X-ray doses ranged from 1 to 11 Gy. At the same time, cells were exposed to either 40Ar (550 MeV/n, 86 keV/micrometers), 28Si (100 MeV/n, 150 keV/micrometers), or 56Fe (115 MeV/n, 442 keV/micrometers) ions. Survival was measured in hamster V79 fibroblasts. Structural aberrations in chromosome 2 were measured by chemical-induced premature chromosome condensation combined with fluorescence in situ hybridization in isolated human lymphocytes. For argon and silicon experiments, measured damage in the mixed radiation field was consistent with the value expected using an additive function for low- and high-LET separated data. A small deviation from a simple additive function is observed with very high-LET iron ions combined to X-rays. PMID- 12530449 TI - Advance directives in dementia research: promoting autonomy and protecting subjects. PMID- 12530450 TI - A few simple truths about your community IRB members. PMID- 12530451 TI - Informed consent for research in schizophrenia.: an alternative for special studies. PMID- 12530452 TI - The intentional tort of patient dumping: a new state cause of action to address the shortcomings of the federal Emergency Medical Treatment and Active Labor Act (EMTALA). PMID- 12530453 TI - Ethics, qualitative research, and ethnography in virtual space. PMID- 12530454 TI - Perception of genetic risk among genetic counselors. AB - A mailed survey of female prenatal genetic counselors, obstetric nurses, and high school biology teachers was conducted to determine if these groups hold different attitudes toward genetic risk and to investigate the extent to which any differences result from the effect of different professional experiences. In this study, the participants were 166 genetic counselors, 116 obstetric nurses, and 78 biology teachers (n=360). Survey participants completed a written questionnaire designed to assess their numeric estimate of the empiric risk for birth defects/genetic problems, their subjective perception of this risk, and their personal use of prenatal diagnosis. Genetic counselors were found to be less likely than the other groups to consider the frequency of birth defects/genetic problems as rare and were 10 times more likely than nurses and 8 times more likely than teachers to have had prenatal diagnosis. Furthermore, more than half of the prenatal diagnosis procedures had by genetic counselors were not medically indicated. These results suggest that genetic counselors have an increased perception of genetic risks relative to nurses or teachers. Possible explanations for this funding are discussed, and the potential role of discordant risk perception in creating biases in genetic counseling process is explored. PMID- 12530455 TI - The Green Journal, 50 years on. PMID- 12530456 TI - Attitudes toward genetic counseling and prenatal diagnosis among a group of individuals with physical disabilities. AB - Few studies have been published on the attitudes of people with physical disabilities toward genetic counseling and prenatal diagnosis. Articles in the lay press and social science literature on this topic, mainly written by disability rights activists and advocates, imply opposition to prenatal diagnosis and the field of clinical genetics by the physically disabled population. In this study, 15 adults with physical disabilities were interviewed regarding their attitudes toward genetic counseling and prenatal diagnosis. Genetic counseling and prenatal diagnosis were generally viewed favorably by this sample of the disability community. Only a small percentage of the sample perceived genetics to be eugenic. Implications for genetic counseling and future research are discussed. PMID- 12530457 TI - Case report: genetic susceptibility testing for breast and ovarian cancer: a patient's perspective. PMID- 12530458 TI - Effect of nocturnal blood pressure reduction on retrobulbar hemodynamics in glaucoma. PMID- 12530459 TI - BRCA1 testing and informed consent in a woman with mild retardation. AB - Cancer predisposition testing can pose complex genetic counseling issues. This case report discusses whether it is appropriate to provide BRCA1 testing to a woman with limited intellectual capacity and documented psychological distress. This case is one of several cancer counseling cases presented to our Genetic Counseling Supervision Group over the past three and a half years. The Supervision Group provided valuable feedback and support to proceed with this challenging case. PMID- 12530460 TI - Effect of nocturnal blood pressure reduction on retrobulbar hemodynamics in glaucoma. PMID- 12530461 TI - Repeated intravitreal injections of triamcinolone acetonide as treatment of progressive exudative age-related macular degeneration. PMID- 12530462 TI - The role of fixation for reliable mfERG results. PMID- 12530463 TI - Case report: resolution of an ethical dilemma. PMID- 12530464 TI - Excitotoxic mechanisms of apoptosis in the mammalian visual system following monocular visual deprivation. PMID- 12530465 TI - Microdialysis sampling of carbamazepine, phenytoin and phenobarbital in subcutaneous extracellular fluid and subdural cerebrospinal fluid in humans: an in vitro and in vivo study of adsorption to the sampling device. AB - The purpose of the study was to determine if binding of the drugs to the sampling equipment during microdialysis would influence the results for carbamazepine, phenytoin and phenobarbital. In vitro experiments with microdialysis catheters and separate parts of catheters were performed to estimate the degree of drug binding to the dialysis equipment. A mathematical model to calculate drug binding and recovery is proposed. In vivo protein unbound carbamazepine concentrations in subcutaneous extracellular fluid at different flow rates (6 patients), unbound carbamazepine (1 patient) and unbound phenobarbital (I patient) in subdural cerebrospinal fluid and subcutaneous extracellular fluid were estimated and the in vivo data were compared to the in vitro results and data generated by the mathematical model. Binding to the soft outlet polyurethane tubing was extensive and variable for phenytoin, which precluded in vivo testing, but limited and more predictable for carbamazepine and phenobarbital. None of the three compounds bound to the hard internaltubing. Phenytoin and phenobarbital did not bind to the dialysis membrane, while a small degree of binding may be present for carbamazepine. In vivo estimates of carbamazepine protein unbound subcutaneous extracellular concentrations by microdialysis, adjusted for binding to the plastic tubing, were 81% of protein unbound plasma concentrations. In single case studies, subdural cerebrospinal fluid and subcutaneous extracellular levels of carbamazepine and phenobarbital were similar and when corrected for binding to the plastic tubings they were also close to protein unbound plasma concentrations. Microdialysis can be used for reliable estimations of protein unbound carbamazepine and possibly phenobarbital concentrations when drug binding to the plastic tubing is considered. Reliable estimation of unbound phenytoin is not possible at present. PMID- 12530466 TI - Sarcolemmal beta-adrenoceptors determined in rat ventricular heart biopsies with (-)-[3H]CGP-12177. AB - Binding of the hydrophilic beta-adrenoceptor ligand (-)-[H]CGP-12177 was investigated by incubating biopsies from rat hearts (left ventricle/interventricular septum) to elucidate the applicability of this approach in determining the content of cell membrane beta-adrenoceptors in heart biopsies. Binding of (-)-[3H]CGP-12177 at 1 nM and 37 degrees reached maximum after 4-10 hr and declined after 10 hr. Binding of (-)-[3H]CGP-12177 at nM and 4 degrees reached equilibrium at 24 hr and was stable up to 96 hr. Total and specific binding was independent of biopsy size in the weight range of 4-35 mg. Competition binding studies with (+)- and (-)-isoprenaline showed that binding was stereospecific. Non-specific binding, determined in the presence of 5 microM (-)-timolol, was 6-8% of total binding at 0.1 nM (at Kd) and 15% of total binding at 1 nM (-)-[3H]CGP-12177. The coefficient of variation for total binding was 5.1%. Dissociation initiated at equilibrium showed complete reversibility of specific binding and was monoexponential with half-life of 0.6 hr at 37 degrees and 30.1 hr at 4 degrees. Binding-saturation experiments at 4 degrees showed beta adrenoceptor density of 7 fmol/mg wet weight and equilibrium dissociation constant of 0.1 nM. Kd calculated from the rate constants of association and dissociation was 0.15 nM. Rapid freezing of tissue in liquid nitrogen with subsequent thawing and binding at 4 degrees C reduced receptor density by 21%. Density of beta-adrenoceptors did not differ in hearts from lean and obese insulin resistant Zucker rats. The results show that the method allows direct determination of sarcolemmal beta-adrenoceptors in small myocardial biopsies at 4 degrees with a minimum of preparation and equipment, using (-)-[3H]CGP-12177 . The method may be useful for other hydrophilic ligands. PMID- 12530467 TI - Amiodarone N-deethylation by CYP2C8 and its variants, CYP2C8*3 and CYP2C8 P404A. AB - Amiodarone is a potent Class III antiarrhythmic drug. The N-deethylation of amiodarone to desethylamiodarone is known to be catalyzed by cytochrome P450 (CYP) 2C8. In the present study, amiodarone N-deethylation by the CYP2C8s, CYP2C8*1 (wild-type), CYP2C8*3, and CYP2C8 P404A (Pro404Ala substitution in exon 8), was investigated by their transient expression in Hep G2 cells. The expression levels of CYP2C8*1 and CYP2C8*3 were similar, whereas the level of CYP2C8 P404A was 55.6% of that of CYP2C8*1. The kinetic parameters of amiodarone N-deethylation were obtained by means of Lineweaver-Burk analysis. The intrinsic clearance (Vmax/Km, per mg of microsomal protein) of amiodarone by CYP2C8 P404A but not CYP2C8*3 was significantly (48.7%) less than that of CYP2C8*1. These results suggest that CYP2C8 P404A but not CYP2C8*3 is less effective in the N deethylation of amiodarone. PMID- 12530468 TI - Nitric oxide synthase/guanylate cyclase pathway modulates the rat vas deferens contractility induced by phenylephrine. AB - The involvement of the nitric oxide synthase/soluble guanylate cyclase pathway on the modulation of phenylephrine-induced contractility in the rat vas deferens was investigated. Phenlylephrine-concentration response curves were obtained in absence and in presence of inhibitors, N(G)-Nitro-L-arginine (L-NOARG), NG-Nitro L-arginine methyl esther (L-NAME) or N(G)-monomethyl-L-arginine (L-NMMA) or GC inhibitior, 1H-(1,2,4)-oxadiaziol-(4,3-a)quinoxalin-1-one (ODQ) or nitric oxide donor, 3-morpholinosydnonimine hydrochloride (SIN-1) alone or together with L NMMA or ODQ. Both nitric oxide synthase and GC inhibitors reduced the Phe-Emax. SIN-1 alone did not change phenylephrine-induced responses and it could reverse the L-NMMA effect but not ODQ effect. The reduction of the phenylephrine-induced contractility obtained in consequence of the inhibition of the nitric oxide/GC pathway suggest that, in the rat vas deferens, despite its well identified relaxant properties, nitric oxide potentiates the contractility induced by adrenergic stimulation. PMID- 12530469 TI - Pien Tze Huang protects the liver against carbon tetrachloride-induced damage. AB - Pien Tze Huang, a traditional Chinese medicine, has been extensively used as a therapeutic drug in the treatment of liver diseases. In this study, we have examined its ability to protect the liver from carbon tetrachloride (CCl4) induced damage in the mouse. Histological observations revealed that CCl4 treatment induced extensive degenerative changes in the hepatocytes surrounding the central veins of the liver. However, these changes were much reduced by more than 28% in mice fed with 0.5 mg of Pien Tze Huang/g body weight/dose (3 doses over 36 hr) prior to CCl4 treatment. The effects of Pien Tze Huang were then further investigated in a hepatoma cell line. Flow analysis showed that it had no significant effects on cell proliferation. When the ability of Pien Tze Huang to influence various response elements of important signal transduction pathways was examined in the hepatoma cell line, it was found that Pien Tze Huang stimulated an increase in the response of AP1, CRE and NFkappaB responsive elements. The transcriptional factors of these responsive elements are known to play important roles in regulating cell death and survival. We thus postulate that the ability of Pien Tze Huang to protect the liver from damage is attained through its ability to modulate the activity of these important signal transduction pathways. PMID- 12530470 TI - The involvement of P-glycoprotein in berberine absorption. AB - Berberine is an important ingredient in a number of traditional Chinese medicines but has been shown to have poor bioavailability in the dog. The aim of this study was to use the P-glycoprotein (P-glycoprotein) inhibitors cyclosporin A, verapamil and the monoclonal antibody C219 in in vivo and in vitro models of intestinal absorption to determine the role of P-glycoprotein in berberine absorption. In the rat recirculating perfusion model, berberine absorption was improved 6-times by P-glycoprotein inhibitors. In the rat everted intestinal sac model, berberine serosal-to-mucosal transport was significantly decreased by cyclosporin A. In Ussing-type chambers, the rate of serosal-to-mucosal transport across rat ileum was 3-times greater than in the reverse direction and was significantly decreased by cyclosporin A. In Caco-2 cells, berberine uptake was significantly increased by P-glycoprotein inhibitors and by monoclonal antibody C219. P-glycoprotein appears to contribute to the poor intestinal absorption of berberine which suggests P-glycoprotein inhibitors could be of therapeutic value by improving its bioavailability. PMID- 12530471 TI - Is cytochrome P450 CYP2D activity present in pig liver? AB - The presence of CYP2D in pig livers has been studied using different strains of pig, different CYP2D test substrates and monoclonal and polyclonal antibodies. The results of the studies lacked consistency, therefore the aim of this study was to identify the reasons for these inconsistencies. Liver microsomes isolated from conventional pigs and minipigs were tested in Western blotting using both monoclonal and polyclonal antibodies against human CYP2D6. The microsomes were also incubated with three different CYP2D tes t substrates.'The immunoblotting only gave a positive response when hybridised with polyclonal antibody. The pig microsomes did not metabolise debrisoquine, but metabolised two other test substrates, dextromethorphan and bufuralol. No correlation was found between the two enzyme assays and CYP2D apoprotein level. On the other hand positive correlations were found between dextromethorphan and bufuralol metabolism and the CYP2B immunochemical protein level, indicating that the CYP2B isoenzyme may be involved in the metabolism of these substrates. Further, assays using immunoinhibition and chemical inhibition of these reactions were performed. No response was obtained in the immunoinhibition assay. When using chemical inhibition, however, an average inhibition percentage of 83 were obtained with orphenadrine, a human CYP2B inhibitor. Average Ki values of 26.9 microM and 43.6 microM for orphenadrine indicate that it was a potent inhibitor. A rat and a mouse CYP2B inhibitor, resveratrol and pilocarpine, inhibited the reaction with an average of 40 and 70 percentage respectively. Orphenadrine did not inhibit CYPIA, CYP2A, CYP2E and CYP3A activities up to more than maximum 12 percentage, showing that it was almost selective for dextromethorphan metabolism. These results indicate that dextromethorphan and bufuralol metabolism may be catalysed by CYP2B and not CYP2D. PMID- 12530472 TI - The chemical UV-filter 3-benzylidene camphor causes an oestrogenic effect in an in vivo fish assay. AB - Chemical UV-filters are used in sun protection products and various kinds of cosmetics. The lipophilic chemical UV-filter 3-benzylidene camphor was investigated for its capability to cause vitellogenin induction, possibly via oestrogen receptor binding, in a well-established in vivo fish assay (juvenile rainbow trout, Oncorhynchus mykiss, vitellogenin ELISA). A clear relationship was demonstrated between the dose of injected 3-benzylidene camphor and the concentration of plasma vitellogenin with a 105-times induction from 68 mg 3 benzylidene camphor /kg/injection and above compared to the control vitellogenin level. The relationship between the injected dose of 3-benzylidene camphor and the percent of responding fish (vitellogenin) was evaluated by logistic regression analysis and effective dose-values (ED-values) were determined. ED10, ED50 and ED90 of 3-benzylidene camphor after 6 days (2 injections) were 6.4, 16 and 26 mg/kg/ injection, respectively. These ED-values place 3-benzylidene camphor among the more potent xenooestrogens discovered to date and necessitates investigations on the distribution, concentration, persistence and bioaccumulation of 3-benzylidene camphor and other UV-filters in nature. PMID- 12530473 TI - Tyrosinase inhibition: general and applied aspects. AB - The active site of tyrosinase is described with a view to depicting its interactions with substrates and inhibitors. Occurrence and mechanism(s) of tyrosinase-mediated browning of agrofood products are reviewed, with regard to both enzymic and chemical reactions, and their control, modulation, and inhibition. Technical and applicational implications are discussed. PMID- 12530474 TI - Quantitative structure-activity relationship study of 2-arylsulfonyl-6 substituted benzonitriles as non-nucleoside reverse transcriptase inhibitors of HIV-1. AB - The reverse transcriptase inhibition of HIV-1, the most common form of HIV, by non-nucleoside 2-arylsulfonyl-6-substituted benzonitriles is analysed through Fujita-Ban and Hansch approaches. The analyses have helped to ascertain the role of different substituents in explaining the observed inhibitory actions of these compounds. From both approaches it appeared that SO2 instead of SO or S at X; and NH2 instead of F at Y (see Figure 1) are advantageous to improving the activity of a compound against HIV-1. This in turn leads to the suggestion that the 2 arylsulfonyl-6-aminobenzonitrile scaffold is the only appropriate structural entity that may further result into potential compounds. Further, the compounds having a OMe substituent at the orthoposition, the bulkier substituents at meta positions and "no" substituent at para-position of 2-arylsulfonyl moiety are beneficial in raising the activity. The two quantitative structure-activity relationship (QSAR) analyses, differing in parametric approach, therefore, provided the grounds for rationalizing the substituent selection in designing more potent compounds of the series. PMID- 12530475 TI - Synthesis and evaluation of some pyrazolo[3,4-d]pyridazinones and analogues as PDE 5 inhibitors potentially useful as peripheral vasodilator agents. AB - A series of pyrazolo[3,4-d]pyridazinones and analogues, potentially useful as peripheral vasodilators, were synthesized and evaluated as inhibitors of PDE5 extracted from human platelets. Several of them showed IC50 values in the range 0.14-1.4 microM. A good activity and selectivity profile versus PDE6 was found for compound 11e (6-benzyl-3-methyl-1-isopropyl-4-phenylpyrazolo[3,4-d] pyridazin 7(6H)-one). Structure-activity relationship studies demonstrated the essential role played by the benzyl group at position-6 of the pyrazolopyridazine system. Other types of pyridazinones fused with five and six membered heterocycles (pyrrole, isoxazole, pyridine and dihydropyridine), as well as some open models were prepared and evaluated. Besides the pyrazole, the best fused systems proved to be isoxazole and pyridine. PMID- 12530476 TI - Characterization of competitive inhibitors for the transferase activity of Pseudomonas aeruginosa exotoxin A. AB - A series of small, nonpolar compounds were tested for their ability to inhibit the ADP-ribosyl transferase activity of Pseudomonas aeruginosa exotoxin A. The IC50 values for the compounds tested ranged from 87 nM to 484 microM for NAP and CMP12, respectively. It was demonstrated that NAP was a competitive inhibitor of the ADPRT reaction for the NAD+ substrate with a Ki of 45 +/- 5 nM, which was in good agreement with the dissociation constant determined independently (KD = 56 +/- 6 nM). The IC50 value for NAP was 87 +/- 12 nM, which strongly correlated with the Ki and KD values. Furthermore, NAP was shown to noncovalently associate with the exotoxin A active site using exhaustive dialysis, NMR, and electrospray mass spectrometry. Finally, a computer molecular model using the X-ray structure of the substrate-bound toxin was generated with NAP bound to the active site of exotoxin A at the nicotinamide-binding site. This model is consistent with the X ray structure of the catalytic domain of poly-ADP-ribose polymerase complexed with 4-amino-naphthalimide (Compound 4) that was included in this study. PMID- 12530477 TI - Inhibition of jack bean urease by 1,4-benzoquinone and 2,5-dimethyl-1,4 benzoquinone. Evaluation of the inhibition mechanism. AB - 1,4-benzoquinone (BQ) and 2,5-dimethyl-1,4-benzoquinone (DMBQ) were studied as inhibitors of jack bean urease in 50 mM phosphate buffer, pH 7.0. The mechanisms of inhibition were evaluated by progress curves studies and steady-state approach to data achieved by preincubation of the enzyme with the inhibitor. The obtained reaction progress curves were time-dependent and characteristic of slow-binding inhibition. The effects of different concentrations of BQ and DMBQ on the initial and steady-state velocities as well as the apparent first-order velocity constants obeyed the relationships of two-step enzyme-inhibitor interaction, qualified as mechanism B. The rapid formation of an initial BQ-urease complex with an inhibition constant of Ki = 0.031 mM was followed by a slow isomerization into the final BQ-urease complex with the overall inhibition constant of Ki* = 4.5 x 10(-5) mM. The respective inhibition constants for DMBQ were Ki = 0.42 mM, Ki* = 1.2 x 10(-3) mM. The rate constants of the inhibitor-urease isomerization indicated that forward processes were rapid in contrast to slow reverse reactions. The overall inhibition constants obtained by the steady-state analysis were found to be 5.1 x 10(-5) mM for BQ and 0.98 x 10(-3) mM for DMBQ. BQ was found to be a much stronger inhibitor of urease than DMBQ. A test, based on reaction with L-cysteine, confirmed the essential role of the sulfhydryl group in the inhibition of urease by BQ and DMBQ. PMID- 12530478 TI - Inhibition of soybean and potato lipoxygenases by bhilawanols from bhilawan (Semecarpus anacardium) nut shell liquid and some synthetic salicylic acid analogues. AB - Bhilawanol diene (3) isolated from bhilawan nut shell liquid was found to be a potent inhibitor of both soybean and potato lipoxygenases with IC50 values of 0.85 microM and 1.1 microM, respectively. However, the monoene (2) and saturated (1) bhilawanols exhibited relatively lower inhibitory activity. In addition, inhibition studies with synthetic analogues of salicylic acid (4-8) suggested that the unsaturated lipophilic side chain may be an absolute requirement for inhibitory activity. PMID- 12530479 TI - Unsymmetrical 1,1'-disubstituted ferrocenes: synthesis of Co(ii), Cu(ii), Ni(ii) and Zn(ii) chelates of ferrocenyl -1-thiadiazolo-1'-tetrazole, -1-thiadiazolo-1' triazole and -1-tetrazolo-1'-triazole with antimicrobial properties. AB - Condensation reactions of 1,1''-diacetylferrocene with different heteroaromatic amines such as, 2-amino-1,3,4-thiadiazole, 5-aminotetrazole and 3-amino-1,2,4 triazole to form unsymmetrically 1,1'-disubstituted ferrocenes have been studied. The obtained compounds have been further investigated for their liganding and biological properties upon chelation with Co(II), Cu(II), Ni(II) and Zn(II) metal ions. The synthesized compounds have been characterized by physical, spectral and analytical data and have been screened against pathogenic bacterial strains e.g., Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus, showing moderate activity as antibacterials in vitro. PMID- 12530480 TI - Arginase activity is inhibited by L-NAME, both in vitro and in vivo. AB - The present study investigated the ability of the arginine analog L-NAME (N(omega)-Nitro-L-arginine methyl ester) to modulate the activity of arginase. L NAME inhibited the activity of arginase in lysates from rat colon cancer cells and liver. It also inhibited the arginase activity of tumor cells in culture. Furthermore, in vivo treatment of rats with L-NAME inhibited arginase activity in tumor nodules and liver, and the effect persisted after treatment ceased. The effect of L-NAME on arginase requires consideration when it is used in vivo in animal models with the aim of inhibiting endothelial NO-synthase, another enzyme using arginine as substrate. PMID- 12530481 TI - Purification and characterization of glucose 6-phosphate dehydrogenase from sheep erythrocytes and inhibitory effects of some antibiotics on enzyme activity. AB - Glucose 6-phosphate dehydrogenase (D-glucose 6-phosphate: NADP+ oxidoreductase, EC 1.1.1.49; G6PD) was purified from sheep erythrocytes, using a simple and rapid method. The purification consisted of three steps; preparation of haemolysate, ammonium sulphate fractionation and 2', 5'-ADP Sepharose 4B affinity chromatography. The enzyme was obtained with a yield of 37.1% and had a specific activity of 4.64 U/mg proteins. Optimal pH, stable pH, molecular weight, and KM and Vmax values for NADP+ and glucose 6-phosphate (G6-P) substrates were also determined for the enzyme. The overall purification was about 1,189-fold. A temperature of +4 degrees C was maintained during the purification process. In order to control the purification of the enzyme SDS polyacrylamide gel electrophoresis (SDS-PAGE) was done in 4% and 10% acrylamide concentration for stacking and running gel, respectively. SDS-PAGE showed a single band for enzyme. Enzymatic activity was spectrophotometrically measured according to Beutler's method at 340 nm. In addition, in vitro effects of gentamicin sulphate, penicillin G potassium, amicasin on sheep red blood cell G6PD enzyme activity were investigated. These antibiotics showed inhibitory effects on enzyme activity. I50 values were determined from Activity%-[Drug] graphs and Ki values and the type of inhibition (noncompetitive) were determined by means of Lineweaver-Burk graphs. PMID- 12530482 TI - Hydatid cyst of the uterus. AB - BACKGROUND: Hydatidosis is a common zoonosis that affects a large number of humans and animals, especially in poorly developed countries. The infesting parasite has four forms named Echinococcus granulosis, E. multilocularis, E. vogeli and E. oligarthrus (very rare in humans). The most frequently involved organs are liver followed by the lung. The involvement of the genital tract is rare and the occurrence in the uterus is an extreme rarity. We report a case of hydatid cyst in the uterus. CASE: A 70-year-old female with a history of hydatid cysts of the liver, was admitted to hospital after complaining of low abdominal pains. On physical and gynecological examinations, no pathological finding was detected. However, the uterus was significantly large for a postmenopausal patient. Transvaginal sonography (TS) revealed a cystic mass in the uterus with a size of 7 x 6 cm. After further examinations a subtotal hysterectomy was performed. Microscopic examination showed scolices of Echinococcus granulosis. CONCLUSION: Hydatid cysts in the genital tract are rare and the occurrence in the uterus is an extreme rarity. Differentiation between hydatid cyst and malignant disease of the related organ is difficult. To avoid misdiagnosis, a careful examination of pelvic masses should be carried out in endemic areas for detection of hydatid cysts. PMID- 12530483 TI - Acyclovir suppression to prevent recurrent genital herpes at delivery. AB - OBJECTIVE: To determine if suppressive acyclovir near term decreased the frequency of clinical recurrences at delivery in women with recurrent genital herpes simplex virus (HSV) infection. METHODS: We conducted a prospective, double blind, randomized trial in 234 women with recurrent genital herpes. Women with genital infection of any frequency were enrolled. Patients received either suppressive oral acyclovir 400 mg three times daily or an identical placebo after 36 weeks' gestation. Clinical lesions were identified, and HSV cultures were obtained at delivery. The frequencies of clinical and subclinical HSV recurrences at delivery were evaluated. RESULTS: Six percent of patients treated with acyclovir, and 14% of patients treated with placebo had clinical HSV at delivery (p = 0.046). No patients in the acyclovir group had positive HSV cultures, compared with 6% of placebo-treated patients (p = 0.029). There was no significant difference in subclinical HSV shedding in the acyclovir group (0%) compared with the placebo-treated group (3%) (p = 0.102). CONCLUSIONS: Suppressive acyclovir therapy significantly decreased the incidence of clinical genital herpes and the overall incidence of HSV excretion at delivery in patients with previous herpes infection. PMID- 12530484 TI - Effect of adjuvant imiquimod 5% cream on sustained clearance of anogenital warts following laser treatment. AB - OBJECTIVES: Imiquimod is an immune response modifier that has demonstrated a good efficacy and relatively low recurrence rates in comparison to other genital wart treatment modalities. The primary objective of this open-label study was to evaluate the effect on sustained clearance of treated lesions and the safety of patient-applied topical imiquimod after laser therapy of external anogenital warts. METHODS: After laser treatment of visible external anogenital warts the ablated region(s) were treated with imiquimod 5% cream three times/week over 12 weeks beginning when the wound healing process was completed, followed by a six month treatment-free observation period for the assessment of sustained clearance of treated lesions. RESULTS: A total of 211 male and female patients was enrolled in the study. After 12 weeks of treatment, 65.4% of all patients showed sustained clearance. During the treatment period, 15 patients (7.1% of 211 patients) presented with recurrent warts in the treated areas, and 58 (27.5%) patients were excluded for other reasons. During the six-month follow-up period, ten additional patients (7.3% of 138 patients) developed wart recurrences. The application of imiquimod 5% cream was well tolerated. The number of patients with adverse events related to study medication declined from the first month of treatment until the end of the third month. Most frequently, mild to moderate itching, burning, pain and erythema were reported. CONCLUSIONS: After laser therapy and sufficient wound healing, administration of imiquimod 5% cream three times/week appears to be safe and to reduce the incidence of wart recurrences. PMID- 12530485 TI - Can the diagnosis of recurrent vulvovaginal candidosis be improved by use of vaginal lavage samples and cultures on chromogenic agar? AB - OBJECTIVE: To investigate if introital and vaginal flushing samples inoculated on chromogenic agar could increase the recovery rate and rapid identification of Candida and non-albicans species, as compared to culture of posterior vaginal fornix samples on Sabouraud agar and speciation of isolates by biochemical tests. METHODS: Samples from the introitus and the posterior vaginal fornix and vaginal lavage samples were collected from 91 women with a history suggestive of recurrent vulvovaginal candidosis (RVVC), and with a suspected new attack of the condition. The specimens were cultured on Sabouraud and CHROMagar. Speciation of yeast isolates was made on the chromogenic agar by API 32C kits and by an atomized system (Vitek). RESULTS: Forty-six (51%) women were positive for Candida from one or more of the samples. The introital cultures were positive in 43 (47%) women, both on Sabouraud and chromogenic agar. From the posterior vaginal fomix, 42 (46%) women were positive on the Sabouraud and 43 (47%) on chromogenic agar cultures, while the vaginal lavage cultures yielded Candida on those two media in 40 (44%) and 41 (45%) cases, respectively. Candida albicans was the most frequent species recovered, from 40 (87%) cases, followed by C. krusei in 4 (9%), C. glabrata in 2 (4%), and C. parapsilosis in one case. There was only one woman who had a mixed yeast infection, by C. albicans and C. krusei. There was only one discrepancy in the speciation as demonstrated by mean of chromogenic agar and API 32C kit. CONCLUSIONS: Neither cultures of introital nor of vaginal lavage samples increases the detection rate of Candida in RVVC cases as compared to cultures of posterior vaginal fornix samples. Use of chromogenic agar is a convenient and reliable means to detect colonization by Candida and differentiate between C. albicans and non-albicans species. PMID- 12530486 TI - Apoptosis in the chorion laeve of term patients with histologic chorioamnionitis. AB - OBJECTIVE: The balance between cell survival and cell death (apoptosis) is critical during development and may affect organ function. Apoptosis is accelerated in the presence of infection and inflammation in a variety of organ systems. The objective of this investigation was to determine if apoptosis was increased in the chorion laeve of term patients with and without histologic chorioamnionitis. METHODS: Records of placental pathology were reviewed with respect to the presence/absence of histologic chorioamnionitis. Sections from formalin-fixed, paraffin-embedded fetal membrane rolls were stained using the TUNEL method. The proportion of apoptotic nuclei was calculated in seven high powered fields/section. Those with and without histologic chorioamnionitis were compared. Data were analyzed using the Mann-Whitney U test, with significance defined as p < 0.05. RESULTS: There was no significant difference in demographic or clinical characteristics between the two groups. The chorion laeve from subjects with histologic chorioamnionitis had significantly more apoptotic nuclei when compared to those without chorioamnionitis (11.2% vs. 5%, p = 0.02). CONCLUSION: Apoptosis is more prevalent in the chorion laeve of fetal membranes with histologic chorioamnionitis. This finding suggests that infection/inflammation may impact cell survival within fetal membranes. The implications of these findings warrant further investigation. PMID- 12530487 TI - Ambiguity and mistakes in the pediatric cardiology vernacular. AB - The pediatric cardiology vernacular is replete with ambiguous and erroneous terminology. This article discusses several examples in a plea to bring the accuracy and precision of the scientific method to the clinical pediatric cardiology vernacular. PMID- 12530488 TI - A multicenter analysis of the choice of initial surgical procedure in tetralogy of Fallot. AB - A current debate on the surgical management of patients with tetralogy of Fallot (TOF) focuses largely on primary repair vs initial shunt in younger and/or smaller patients. To characterize practice patterns throughout the United States with respect to the choice of initial surgical procedure in patients with TOF, we analyzed data from a multicenter database. This retrospective study focused on 938 patients from 12 institutions who underwent their initial operation during the 10-year period 1986 through 1995. Overall, the percentage of shunts decreased from 35.1% (1986-1990) to 22.0% (1991-1995) (p < 0.0001). The percentage of primary repairs increased accordingly. However, there was marked interinstitutional variability. For the group of patients aged 3 months or less the overall in-hospital mortality was significantly higher than that for older patients for both shunts and repairs. Multiple logistic regression analysis indicated that age, weight, date of surgery, and the interactions between date of surgery and institutional volume and between age and institutional volume were significant predictors of the initial surgical management of TOF. With this model only part of the observed variance could be explained. Other unidentified variables, including "institutional preference," may be significant factors influencing the choice of initial surgical procedure. PMID- 12530490 TI - Test characteristics of a level I or II fetal ultrasound in detecting structural heart disease. AB - At the University of California Davis Medical Center, a screening fetal ultrasound examination (level I or II) incorporates a comprehensive segmental evaluation of the fetal heart. This study evaluated the reliability of the fetal ultrasound exam in the detection of abnormal heart anatomy. Our retrospective study reviewed results of 614 antenatal patients that had a screening fetal ultrasound exam. All patients subsequently underwent a detailed targeted fetal cardiac ultrasound exam performed by a pediatric cardiac sonographer and reviewed by a board-certified pediatric cardiologist. Of these 614 patients, 60 fetuses had structural heart disease by the targeted fetal exam. The screening fetal ultrasound exam correctly identified 55 of the 60, with 5 false negatives (8.3% false-negative rate) and 1 false positive (1.7% false-positive rate). Our study suggests that if a screening fetal ultrasound exam incorporates a segmental evaluation of the fetal heart it can reliably detect abnormal heart anatomy. At our institution a targeted fetal cardiac exam is now used to confirm and provide detailed assessment of the heart anatomy when a screening fetal exam is positive for heart disease. PMID- 12530489 TI - Immunologic profile of patients with protein-losing enteropathy complicating congenital heart disease. AB - The immunologic profile of patients with congenital heart disease complicated by protein-losing enteropathy (PLE) is undefined. The aim of this study was to assess the lymphocyte subpopulation and immunglobulin (Ig) pattern in patients with PLE complicating congenital heart disease. The immunologic profile of six patients with congenital heart disease complicated by PLE was compared to that of controls without PLE matched for age and cardiac interventions. Enteric protein loss was documented by Tc99m-labeled albumin scintigraphy. The lymphocyte subpopulations were enumerated using flow cytometry, whereas serum IgG, IgA, and IgM concentrations were measured by the turbidimetric technique. The cardiac diagnoses included complex cyanotic heart disease post-Fontan procedure (n = 3), and one each of tetralogy of Fallot, restrictive cardiomyopathy, and valvar pulmonary stenosis. In patients with PLE, the T lymphocyte (CD3+) count was significantly lower (300 +/- 186 vs 2070 +/- 1171/microl, p = 0.017); both the helper/inducer lymphocytes (CD4+) (127 +/- 158 vs 927+/- 377/microl, p = 0.006) and suppressor/cytotoxic lymphocytes (CD8+) (129 +/- 49 vs 850 +/- 695/microl, p = 0.057) reduced with reversal of CD4(+)/CD8(+) ratio (0.81 +/- 0.68 1.64 +/- 0.89, p = 0.027). Furthermore, IgG level was significantly reduced (5.12 +/- 2.84 vs 12.5 +/- 1.58 g/L, p = 0.005) and IgA level tended to be lower (1.36 +/- 1.37 vs 2.50 +/- 0.80 g/L, p = 0.095). In contrast, the B lymphocyte (CD19+) count (340 +/- 151 vs 618 +/- 427/microl, p = 0.25), natural killer cell count (CD16(+) 56(+) CD3(-)) (252 +/- 212 vs 276 +/- 251/microl, p = 0.85), and IgM level (0.98 +/- 0.59 vs 1.12 +/- 0.25 g/L, p = 0.67) were similar for both groups. None of the patients developed opportunistic or severe viral infections. Abnormal immunologic profile of both the cellular and humoral arms of the immune system occurs in patients with congenital heart disease complicated by PLE. Nonetheless, these abnormalities perhaps appear quantitative rather than qualitative in nature, although further functional studies of antibody production and lymphocyte proliferation assays are required to support this proposition. PMID- 12530491 TI - The incidence of arrhythmias in a pediatric cardiac intensive care unit. AB - A pediatric cardiac intensive care unit (CICU) manages critically ill children and adults with congenital or acquired heart disease. These patients are at increased risk for arrhythmias. The purpose of this study was to prospectively evaluate the incidence of arrhythmias in a pediatric CICU patient population. All patients admitted to the CICU at the Cardiac Center at The Children's Hospital of Philadelphia between December 1, 1997, and November 30, 1998, were evaluated prospectively from CICU admission to hospital discharge via full disclosure telemetry reviewed every 24 hours. Arrhythmias reviewed included nonsustained and sustained ventricular tachycardia (VT), nonsustained and sustained supraventricular tachycardia (SVT), atrial flutter and fibrillation, junctional ectopic tachycardia, and complete heart block. We reviewed 789 admissions consisting of 629 patients (age range, 1 day-45.5 years; median, 8.1 months). Hospital stay ranged from 1 to 155 days (total of 8116 patient days). Surgical interventions (n = 602) included 482 utilizing cardiopulmonary bypass. During the study period, there were 44 deaths [44/629 patients (7.0%)], none of which were directly attributable to a primary arrhythmia. The operative mortality was 5.1%. Overall, 29.0% of admissions had one or more arrhythmias the most common arrhythmia was nonsustained VT (18.0% of admissions), followed by nonsustained SVT (12.9% of admissions). Patients admitted to a pediatric CICU have a high incidence of arrhythmias, most likely associated with their underlying pathophysiology and to the breadth of medical and surgical interventions conducted. PMID- 12530492 TI - QT dispersion in infants with apparent life-threatening events syndrome. AB - Apparent life-threatening event (ALTE) is a term used to define an event of unknown cause after an infant is found limp, cyanotic, bradycardic, and/or requires resuscitation. Eight to 15% of children with ALTE die of sudden infant death syndrome. Obstructive sleep apnea, bradycardia, gastroesophageal reflux, and laryngotracheal abnormalities are frequently associated with ALTE. Wide QT dispersion is associated with sudden death in heart failure and increased risk of ventricular fibrillation in acute myocardial infarction. Here, we assess QT dispersion in infants with ALTE and its correlation to clinical and electrocardiographic indices. The study included eighty nine infants (age 2.14 +/ 1.8 months, 46 males and 43 females) referred with ALTE to the pediatric emergency room and 18 controls (age 2.77 +/- 2.2 months) who underwent electrocardiogram assessment of QTmin, QTmax, QT dispersion (QT-D), and as well as QTmin, QTmax, and QT-D corrected for heart rate (QTcmin, QTcmax, QTC-D, respectively). All infants were referred at the usual diagnostic tests-the gastroesophageal reflux test, apnea monitoring, Holter ECG monitoring, electroencephalogram, and Doppler echocardiography. QT-D, QTc-D, and QTc-min were significantly greater in the ALTE group (p < 0.01). Greater QTc-D was found in males compared to females (p < 0.001). QT-D and QTc-D showed little or no correlation with age of infant or positivity of diagnostic tests. QTc has been found by multiple regression analysis to be the independent variable with the greatest impact on QTc-D (beta = -0.68, p < 0.001). PMID- 12530493 TI - Physical fitness of urban American children. AB - The Bruce treadmill protocol was used to evaluate the exercise endurance in a pediatric urban population. The sample population consisted of 525 healthy children 4 to 18 years of age (303 males and 222 females). Endurance time increased with age in males and increased up to 10 to 12 years of age in females, thereafter leveling off. Results were compared with those of a widely used standard reference of normal for treadmill exercise testing in children. Mean endurance times were significantly shorter compared to those of the reference study (p < 0.03). When the subjects were grouped in percentiles based on the reference study, 61% of males and 81% of females performed below the 25th percentile. There was a strong negative correlation between body mass index and endurance time (p < 0.0001), suggesting that obesity is a major contributor to decreased physical fitness. Compared to subjects in the reference study, children from a large U.S. metropolitan area have markedly diminished exercise endurance times, suggesting poor physical fitness. Inactivity with consequent lack of fitness, if unchecked, are likely to lead to cardiovascular disorders in adulthood. Efforts should be made to encourage physical fitness in children. PMID- 12530494 TI - Effect of dobutamine on a Doppler echocardiographic index of combined systolic and diastolic performance. AB - Dobutamine stress echocardiography has become accepted in the evaluation of cardiac functional reserve. Although the Doppler-derived index of combined systolic/diastolic myocardial performance (Tei index) has been reported to be easily obtainable and useful for predicting left ventricular performance, the effect of dobutamine on the Tei index has not been determined in children. To assess the effect of dobutamine on the Tei index, 8 patients who had undergone surgery for ventricular septal defect and 7 patients who had a history of Kawasaki disease were examined. Echocardiographic recordings were obtained before and after dobutamine infusion (5 microg/kg per minute). Variables measured were transmitral flow velocities (E, A, E/A), rate-corrected mean velocity of circumferential fiber shortening (rate-corrected Vcf), and IMP. We measured isovolumic contraction time (ICT), isovolumic relaxation time (IRT), and ejection time (ET) and then calculated the Tei index using the following formula: Tei index = (ICT + IRT)/ET. Dobutamine infusion increased rate-corrected Vcf (29%, p < 0.01), peak E (7%, p < 0.05), and peak A (13%, p < 0.05). E/A ratio did not change during dobutamine infusion. ET, ICT, and IRT were found to decrease during dobutamine infusion. The magnitude of the change in the ICT (-21%, p < 0.01) was greater than those in IRT (-12%, p < 0.01) and ET (-8%, p < 0.05). The decrease in Tei index (-16%, p < 0.01) resulted in a decrease in ICT/ET (-21%, p < 0.01) and a slight decrease in IRT/ET (-12%, p < 0.05). The Tei index has potential as a sensitive indicator of the effects of inotropic stimilation on global left ventricular function. PMID- 12530495 TI - Afterload reduction therapy in patients following intraatrial baffle operation for transposition of the great arteries. AB - Patients with intraatrial baffle procedure for transposition of the great arteries (TGA) have diastolic dysfunction, decreased exercise capacity, stroke volume response and elevated systemic vascular resistance (SVR) during exercise. Angiotensin-converting enzyme (ACE) inhibitors improve exercise capacity in adults with congestive heart failure by improving diastolic function and decreasing SVR. We tested the hypothesis that ACE inhibitors decrease SVR and improve exercise capacity in patients after intraatrial baffle procedure for TGA. We studied the effects of enalapril in nine patients with TGA s/p intraatrial switch (mean age, 13.8 +/- 3 years) 7 to 21 years (mean, 12 +/- 4 years) after intraatrial baffle procedure. Enalapril (0.5 mg/kg/day, maximum dosage 20 mg bid) was administered for 12 months. Patients exercised using a cycle ergometer ramp protocol (0.25 W/kg/min) before enalapril (baseline), 1 month, 6 months, and 12 months after treatment initiation. Heart rate, blood pressure, cardiac output, respiratory rate, minute ventilation, oxygen consumption (VO2), total exercise time, work, and power were measured. SVR, cardiac index, and stroke volume index (SVI) were calculated. Two-tailed paired Student's t-test was used to compare data to those of normal control patients and the patients' baseline data. Patients had lower resting heart rate, cardiac index, maximum heart rate, cardiac index (CI), SVI, VO2, exercise time, work, and power and higher maximal SVR at baseline compared to normal control patients. There was no significant difference in total exercise time, work, power, VO2 (rest/peak), SVR, SVI, and CI after 12 months of therapy compared to patients' baseline values. We conclude that short term (<1 year) use of enalapril does not improve exercise performance in patients with TGA in whom the intraatrial baffle procedure has been performed. PMID- 12530496 TI - Perioperative care of the adult with congenital heart disease in a free-standing tertiary pediatric facility. AB - Adult patients with congenital heart disease are presenting more frequently for cardiac surgery. Frequently, pediatric congenital heart surgeons perform these procedures at pediatric hospitals. Between July 1995 and June 2000, a retrospective review of adult patients (> or = 18 years old) who had undergone cardiothoracic operations was performed. A total of 112 operations were performed and divided into two groups--81 cardiac operations in 79 patients and 31 noncardiac operations in 23 patients. One patient had a cardiac and noncardiac operation performed. The overall early operative mortality was 6% (6/101). There were 3 late deaths. New-onset cardiac arrhythmias requiring treatment were diagnosed after 5/81 (6%) cardiac operations. Six of 79 (7%) patients were diagnosed with postoperative clinical depression. An acceptable mortality can be achieved when adult patients undergo cardiothoracic operations at a pediatric facility. New-onset arrhythmias necessitating treatment are relatively common, and postoperative clinical depression should be anticipated. PMID- 12530497 TI - Unusual cardiac malformations in conjoined twins: thoracopagus twins with conjoined pentalogy of Cantrell and an omphalopagus twin with atretic ventricles. AB - Two unrelated cases of conjoined twins were found to have cardiac malformations that apparently have not been reported previously. In one case, thoracopagus twins had an extensive thoracoabdominal wall defect that resulted in ectopia cordis of a conjoined heart along with evisceration of the shared liver and intestine along with one spleen. These malformations, accompanied by defects in the sternum, diaphragm, and supraumbilical abdominal wall, constitute a conjoined pentalogy of Cantrell. In the second case, the heart of one of omphalopagus twins consisted of a solid ventricular mass with only a minute aortic cavity but no atrioventricular communication-an ineffective heart that could develop only in a conjoined or chorioangiopagus twin. In both cases, a common atrium lay in the primitive (embryologic) position caudal to the ventricles. PMID- 12530498 TI - A national survey on the pediatric cardiologist's clinical approach for patients with Kawasaki disease. AB - In 1994, the American Heart Association (AHA) published the most recent guidelines for long-term cardiovascular management of Kawasaki disease. Since then, recent publications have shed new light on different diagnostic, prognostic, and management issues. We sought the opinion of pediatric cardiologists practicing in U.S. fellowship programs on the subject by means of a multiple-choice survey. Two questions addressed therapy in the acute phase, each preceded by a statement from related literature. Ten duplicate questions addressed the long-term cardiovascular management in five sets of paired questions; each question was first given in reminiscence of a clinical situation and then preceded by a statement from particular publications representative of new information that has become available since the publication of the 1994 AHA guidelines. All questions were provided in the same mailing. Replies were received from 97 participants practicing at 29 institutions. For the acute illness, 21% of respondents do not use high-dose aspirin, and 50% support reassessment of current guidelines. Universal intravenous immune globulin (IVIG) administration is followed by 97%, among whom 20% agree that evaluation of selection criteria is needed. For long-term management, 60-75% advocate regular follow-up of risk level I patients, and 80% favor periodic follow-up, with stress imaging (34-40%), for risk level II. For risk level IV more respondents favor stress echocardiography as opposed to nuclear imaging, in consonance with recent literature. For risk levels III and IV, 36-40% perform coronary angiography on a regular basis, whereas 60% do so when coronary symptoms are present or when stress imaging suggests myocardial ischemia. Finally, 19-25% of respondents do not routinely advise healthy lifestyle to patients free of coronary artery lesions. In conclusion, the guidelines for conventional therapy in the acute phase and long-term cardiovascular management need to be revised. PMID- 12530499 TI - Functional pulmonary atresia in a newborn with normal intracardiac anatomy. AB - Functional pulmonary atresia is a relatively rare clinical condition usually associated with Ebstein's malformation, tricuspid valve dysplasia, Uhl's anomaly, or transient myocardial ischemia with severe tricuspid regurgitation. The occurrence of functional pulmonary atresia associated with transient tricuspid regurgitation in a newborn with an anatomically normal heart is even more uncommon. We describe a case in which color Doppler flow mapping played an essential role in diagnosis and follow-up of this clinical condition in a newborn who had normal intracardiac anatomy. PMID- 12530500 TI - Successful combined orthotopic liver transplant and transcatheter management of atrial septal defect, patent ductus arteriosus, and peripheral pulmonic stenosis in a small infant with Alagille syndrome. AB - We report on a 4-kg infant with Alagille syndrome and congenital heart disease consisting of atrial septal defect (ASD), patent ductus arteriosus (PDA), and severe peripheral pulmonic stenosis. He underwent successful orthotopic liver transplant along with catheter closure of the ASD and PDA using the Amplatzer device and placement of Palmaz stents in both pulmonary arteries. PMID- 12530501 TI - An aberrant umbilical vein draining into the coronary sinus. PMID- 12530502 TI - Fetal diagnosis and postnatal follow-up of an asymptomatic congenital left ventricular diverticulum. PMID- 12530503 TI - Pulmonary varix secondary to pulmonary vein obstruction. PMID- 12530504 TI - Feasibility and limitations of acridine orange fluorescence technique using a Malaria Diagnosis Microscope in Myanmar. AB - We studied parasite detectability in thick films by an acridine orange fluorescence technique (AO) to test its applicability and the use of a Malaria Diagnosis Microscope (MDM)-ESL in the detection of parasites, compared to the conventional Giemsa staining method. This study was conducted on 1,390 clinically suspected malaria cases of Thaton township, Myanmar. We found sensitivities of 82.8% for Plasmodium falciparum (P. falciparum) and 100% for Plasmodium vivax (P. vivax) and specificities of 97.1% for P. falciparum and 98.6% for P. vivax. AO had a higher sensitivity than Giemsa-stained films at low levels of parasitemia (< 1,000/microl). AO showed lower sensitivity and higher specificity than the Giemsa method at parasite levels of more than 1,000/microl. The results of using the AO method, achieved by both novice and experienced observers, showed no significant difference and required less practice to perform the test as well as to identify the parasite. The acridine orange fluorescence technique using a malaria diagnosis microscope MDM-ESL series is simple, rapid and cost effective. The microscope is conveniently operable using standard AC power or a 12-V DC car battery, and it is easily convertible to a conventional biological microscope. With the exception of species differentiation, which is not possible with this method, this method would be appropriate for both clinical and epidemiological studies. PMID- 12530505 TI - Camptothecin induces urokinase-type plasminogen activator gene-expression in human RC-K8 malignant lymphoma and H69 small cell lung cancer cells. AB - We previously reported that anthracyclines, which could generate reactive oxygen species (ROS), could induce the urokinase-type plasminogen activator (uPA) gene expression in human RC-K8 malignant lymphoma cells and in H69 small cell lung cancer (SCLC) cells. In screening other uPA-inducible anti-cancer agents, we found that camptothecin (CPT) and its derivative, SN38, could induce uPA in RC-K8 and H69 cells. CPT and SN38, which are also used for the treatment of lymphoma and SCLC, significantly increased the uPA accumulation in the conditioned media of both cells in a dose-dependent manner. The maximum induction of uPA mRNA levels was observed 24 h after stimulation. Pretreatment with pyrrolidine dithiocarbamate (PDTC), an anti-oxidant, inhibited the CPT-induced uPA mRNA expression. Thus, CPT induces uPA through gene expression, and, therefore, CPT may influence the tumor-cell biology by up-regulating the uPA/plasmin system. PMID- 12530506 TI - Evaluation of a method for typing the microsatellite D12S391 locus using a new primer pair and capillary electrophoresis. AB - We describe a modified method for typing a polymorphic microsatellite D12S391 locus by PCR using a newly designed primer pair. This primer pair produces shorter D12S391 amplified fragments (104-156 bp) than the primer pair originally described by Lareu et al. (209-261 bp). The detection system for the D12S391 locus using the new primer pair and capillary electrophoresis (CE) analysis was evaluated using various forensic samples. The typing results from 70 DNA samples using the new primer pair and the original primer pair were completely identical. One hundred twenty-five amplified fragments from D12S391 alleles were sized correctly within +/- 0.25 bp of the D12S391 allelic ladder. A rare allele, 19.3, previously found only in Caucasians, was found for the first time in a Japanese subject, and it was clearly distinguished from allele 20 by the CE analysis. This detection system was sensitive and could detect D12S391 types from 16 pg of genomic DNA, and from a minor component at a ratio of 1:10 in mixed samples. This system was more useful for the analysis of degraded DNA than was the method using the original primer pair, and could detect D12S391 types from bloodstains that had been stored for 26 years. In addition, the specificity of the method was demonstrated using nonhuman DNA. PMID- 12530507 TI - Diabetic gastroparesis in association with autonomic neuropathy and microvasculopathy. AB - Gastroparesis is a frequent and sometimes life-threatening complication of diabetes mellitus. Autonomic neuropathy seems to be one of the most important mechanisms underlying this entity, together with the other probable pathologies. The present study was performed in order to identify an alternative to gastric scintigraphy as a screening test. The gastric emptying times of 60 subjects (Group 1: 20 insulin-dependent patients, Group 2: 20 non-insulin-dependent diabetes mellitus patients, and Group 3: 20 healthy volunteers) were monitored by gastric scintigraphy. Perception thresholds for cold, heat, and vibration were tested by a quantitative sensory test, and QTc dispersions were calculated from standard electrocardiography recordings. In addition, fasting blood glucose, hemoglobin A1c and urine beta2-microglobulin and microalbumin concentrations were determined for the patient groups. Funduscopic examination was performed by an independent ophthalmologist. Gastroparesis was determined in both patient groups, regardless of fasting blood glucose and hemoglobin A1c concentrations. A strong correlation was observed between nephropathy, retinopathy, and cardiac autonomic denervation (QTc) and gastroparesis. In conclusion, retinal and renal microvasculopathy parameters and cardiac autonomic function tests may be useful for screening diabetic patients for gastroparesis. PMID- 12530508 TI - Hepatopancreatic arterial ring: bilateral symmetric typology in human celiaco mesenteric arterial system. AB - The celiac and mesenteric arterial system including the left gastric, splenic, common hepatic, and superior mesenteric arteries shows various types of origins, courses, ramifications and anastomoses. In order to explain the various expressions of this system, we have proposed a typological model, in which celiacomesenteric arteries develop as paired or bilaterally symmetrical primordial vessels originated from the anterior aspect of the aorta, and these vessels anastomose each other with longitudinal and horizontal pathways. Here, we report 3 unusual cases characterized by arterial rings, formed by the left gastric, left accessory hepatic, proper hepatic, anterior pancreaticoduodenal, and dorsal pancreatic arteries. The dorsal pancreatic and anterior pancreaticoduodenal arteries are located to the right and left of the embryonic pancreas developing in the dorsal mesentery, respectively. Such hepatopancreatic arterial rings simultaneously containing right and left elements can only be explained using our typological model, in which the concept of paired arteries or bilateral symmetry is introduced. PMID- 12530509 TI - Obesity and the risk of diabetes mellitus in middle-aged Japanese men. AB - The morbidity of diabetes mellitus is increasing gradually in Japanese populations. It is important to clarify the risk factors of diabetes in Japanese populations in order to take adequate measures against the increasing morbidity of diabetes. In order to evaluate the link between past and concurrent obesity and diabetes in middle-aged Japanese men, we conducted a worksite-based historical cohort study in Okayama, Japan in 1999. Annual health examination data of middle-aged male workers in a worksite were collected. The relative risks of past and concurrent obesity for developing diabetes were calculated. Subjects with a past history of obesity at between 40 and 50 years of age had a significantly higher risk of developing diabetes by age 55 than did subjects in the normal weight group. These results suggest that, in order to prevent diabetes in middle-aged Japanese men, health guidance for normal weight maintenance should be provided not only for middle-aged men, but also for men under age 40. PMID- 12530510 TI - Daily low-dose cisplatin and concurrent thoracic irradiation for poor-risk patients with unresectable non-small-cell lung cancer. AB - A pilot study was conducted to assess the efficacy and feasibility of daily low dose cisplatin with concurrent thoracic irradiation for clinically unresectable non-small-cell lung cancer (NSCLC). Patients with inoperable NSCLC who had poor risk factors such as advanced age, poor performance status, poor lung function, or concomitant active malignancy were entered into the study. Low-dose cisplatin (6 mg/m2) was administered daily before concurrent thoracic irradiation (2 Gy/day; total dose of 60 Gy) was given. Twenty-five patients were registered. The majority of the patients had either stage IIIA (24.0%) or stage IIIB (60.0%) disease. Fifteen patients (60.0%) completed the planned treatment. Both chemotherapy and radiotherapy were stopped in 3 patients (12.0%) due to poor response, and 7 patients (28.0%) partly received radiotherapy alone as a result of their toxicity response. The proportion of total administered dose to planned dose was 90.9% for chemotherapy and 99.3% for radiotherapy, which were comparable to those in previous studies for LA-NSCLC patients without poor risk factors. Grade 3 leukopenia and neutropenia developed in 14 patients (56.0%) and 10 patients (40.0%), respectively, but grade 4 toxicity was not encountered. Grade 3 pneumonitis and esophagitis were observed in 4 patients (16.0%) and 2 patients (8.0%), respectively. The overall response rate was 60.0%. The median survival time was 22 months, and the 2-year survival rate was 50.3%. Daily low-dose cisplatin and concurrent thoracic irradiation were well tolerated even by poor risk patients with NSCLC, and showed a therapeutic efficacy similar to that for good-risk patients. PMID- 12530511 TI - Anterior intercostobrachial nerve penetrating the pectoralis minor or major muscle. AB - Two previously unknown anomalies of the anterior intercostobrachial nerve were described. In one case, the anterior intercostobrachial nerve penetrated the pectoralis minor muscle. In the other case, it penetrated the pectoralis major muscle. In both cases, the anomalous nerve supplied the skin of the upper arm. PMID- 12530512 TI - A new view concerning an actomyosin motor. PMID- 12530513 TI - Statins: the new aspirin? AB - 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, or statins, have been described as the principal and the most effective class of drug to reduce serum cholesterol levels. Statin therapies have been shown to reduce cardiovascular events, including myocardial infarction, stroke, and death, significantly, by altering vascular atherosclerosis development in patients with or without coronary artery disease symptoms. Extensive use of statins has led to the increase of some undesirable effects that are heavily counterbalanced by the benefits. Indeed, pleiotropic effects extend far beyond cholesterol reduction and involve non-lipid-related mechanisms that modify endothelial functions, immunoinflammatory responses, smooth muscle cell activation, proliferation and migration, atherosclerotic plaque stability, and thrombus formation. In this review, we describe in detail the targets and mechanisms of action of statins. PMID- 12530514 TI - Sensing life: regulation of sensory neuron survival by neurotrophins. AB - Neurotrophins are a family of structurally and functionally related neurotrophic factors which, in mammals, include: nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3 (NT-3), and NT-4/5. In addition to their canonical role in promoting neuronal survival, these molecules appear to regulate multiple aspects of the development of the nervous system in vertebrates, including neuronal differentiation, axon elongation and target innervation, among others. Actions of neurotrophins and of their receptors in vivo are being analyzed by loss-of-function or gain-of-function experiments in mice. Here, we review the phenotypes of the primary sensory system in these mutant mouse strains and the different strategies specifically involved in the regulation of neuronal survival by neurotrophins in this portion of the nervous system. PMID- 12530515 TI - Mitochondria as the conductor of metabolic signals for insulin exocytosis in pancreatic beta-cells. AB - Mitochondrial metabolism is crucial for the coupling of glucose recognition to the exocytosis of the insulin granules. This is illustrated by in vitro and in vivo observations discussed in the present review. Mitochondria generate ATP, which is the main coupling messenger in insulin secretion. However, the subsequent Ca2+ signal in the cytosol is necessary but not sufficient for full development of sustained insulin secretion. Hence, mitochondria generate ATP and other coupling factors serving as fuel sensors for the control of the exocytotic process. Numerous studies have sought to identify the factors that mediate the amplifying pathway over the Ca2+ signal in glucose-stimulated insulin secretion. Predominantly, these factors are nucleotides (GTP, ATP, cAMP, NADPH), although metabolites have also been proposed, such as long-chain acyl-CoA derivatives and glutamate. Hence, the classical neurotransmitter glutamate receives a novel role, that of an intracellular messenger or co-factor in insulin secretion. This scenario further highlights the importance of glutamate dehydrogenase, a mitochondrial enzyme well recognized to play a key role in the control of insulin secretion. Therefore, additional putative messengers of mitochondrial origin are likely to participate in insulin secretion. PMID- 12530516 TI - Molecular aspects of membrane fission in the secretory pathway. AB - Membrane fission is essential in various intracellular dissociative transport steps. The molecular mechanisms by which endocytic vesicles detach from the plasma membrane are being rapidly elucidated. Much less is known about the fission mechanisms operating at Golgi tubular networks; these include the Golgi transport and sorting stations, the trans-Golgi and cis-Golgi networks, where the geometry and physical properties of the membranes differ from those at the cell surface. Here we discuss the lipid and protein machineries that have so far been related to the fission process, with emphasis on those acting in the Golgi complex. PMID- 12530517 TI - The origin, diversity, and structure function relationships of insect luciferases. AB - Luciferases are the enzymes that catalyze the reactions that produce light in bioluminescence. Whereas the oxidative mechanism which leads to light emission is similar for most luciferases, these enzymes and their substrates are evolutionarily unrelated. Among all bioluminescent groups, insects constitute one of the most diverse in terms of biochemistry. In the fungus-gnats (Mycetophilidae: Diptera), for example, bioluminescence is generated by two biochemically distinct systems. Despite the diversity, investigations on insect luciferases and biochemistry have been conducted mostly with fireflies. The luciferases from the related phengodid beetles, which can produce green to red bioluminescence using the same chemistry as firefly luciferases, have been recently investigated. Beetle luciferases originated from ancestral acyl-CoA ligases. Present data suggest that conserved motifs among this class of ligases are involved in substrate adenylation. The three-dimensional structure of firefly luciferase was recently solved and mutagenesis studies have been performed identifying putative residues involved in luciferin binding and bioluminescence color determination in several beetle luciferases. The knowledge gained through these studies is helping in the development of useful reporter gene tools for biotechnological and biomedical purposes. PMID- 12530518 TI - Myelin sheaths: glycoproteins involved in their formation, maintenance and degeneration. AB - Myelin sheaths are formed around axons by extending, biochemically modifying and spiraling plasma membranes of Schwann cells in the peripheral nervous system (PNS) and oligodendrocytes in the central nervous system (CNS). Because glycoproteins are prominent components of plasma membranes, it is not surprising that they have important roles in the formation, maintenance and degeneration of myelin sheaths. The emphasis in this review is on four integral membrane glycoproteins. Two of them, protein zero (P0) and peripheral myelin protein-22 (PMP-22), are components of compact PNS myelin. The other two are preferentially localized in membranes of sheaths that are distinct from compact myelin. One is the myelin-associated glycoprotein, which is localized at the inside of sheaths where it functions in glia-axon interactions in both the PNS and CNS. The other is the myelin-oligodendrocyte glycoprotein, which is preferentially localized on the outside of CNS myelin sheaths and appears to be an important target antigen in autoimmune demyelinating diseases such as multiple sclerosis. PMID- 12530520 TI - Role of glutamic acid decarboxylase in the pathogenesis of type 1 diabetes. AB - Glutamic acid decarboxylase (GAD) is considered to be one of the strongest candidate autoantigens involved in triggering beta-cell-specific autoimmunity. The majority of recent onset type 1 diabetes patients and pre-diabetic subjects have anti-GAD antibodies in their sera, as do nonobese diabetic (NOD) mice, one of the best animal models for human type I diabetes. Immunization of young NOD mice with GAD results in the prevention or delay of the disease as a result of tolerizing autoreactive T cells. Autoimmune diabetes can also be prevented by the suppression of GAD expression in antisense GAD transgenic mice backcrossed with NOD mice for seven generations. These results support the hypothesis that GAD plays an important role in the development of T-cell-mediated autoimmune diabetes. However, there is some controversy regarding the role of GAD in the pathogenesis of diabetes. Whether GAD truly plays a key role in the initiation of this disease remains to be determined. The examination of the development of insulitis and diabetes in beta-cell-specific GAD knockout NOD mice will answer this remaining question. PMID- 12530519 TI - Role of hydrogen peroxide and oxidative stress in healing responses. AB - Oxidative stress is a host defense mechanism whose involvement in maintaining homeostasis and/or inducing disease has been widely investigated over the past decade. Various reactive oxygen species (ROS) have been defined and the enzymes involved in generating and/or eliminating them have been widely studied. In this review we briefly discuss general mechanisms of oxidative stress and the oxidative stress response of the host. We focus primarily on hydrogen peroxide and summarize the systems involved in its formation and elimination. We describe mechanisms whereby hydrogen peroxide and other ROS can modify protein conformation and, thus, alter protein function, and describe a group of transcription factors whose biological activity is modulated by the redox state of cells. These basic aspects of oxidative stress are followed by a discussion of mechanisms whereby hydrogen peroxide and other ROS can modulate some physiological and pathological processes, with special emphasis on wound healing and scarring of the liver. PMID- 12530521 TI - Bacterial cold-shock proteins. AB - Members of a family of small cold-shock proteins (CSPs) are induced during bacterial cell response to a temperature decrease. Here we review available data about the structure, molecular properties, mechanism of induction and possible functions of CSPs. CSPs preferentially bind single-stranded RNA and DNA and appear to play an important role in cell physiology under both normal and cold shock conditions. Although the function of CSPs in cold-shock adaptation has not yet been elucidated in detail, a number of experimental evidences suggests that CSPs bind messenger RNA (mRNA) and regulate ribosomal translation, rate of mRNA degradation and termination of transcription. PMID- 12530522 TI - Cellular and molecular aspects of drugs of the future: oxaliplatin. AB - Oxaliplatin (Eloxatine) is a third-generation platinum compound which has shown a wide antitumour effect both in vitro and in vivo, a better safety profile than cisplatin and a lack of cross-resistance with cisplatin and carboplatin. In this scenario, oxaliplatin may represent an innovative and challenging drug extending the antitumour activity in diseases such as gastrointestinal cancer that are not usually sensitive to these coordination complexes. Oxaliplatin has a non hydrolysable diaminocyclohexane (DACH) carrier ligand which is maintained in the final cytotoxic metabolites of the drug. Like cisplatin, oxaliplatin targets DNA producing mainly 1,2-GG intrastrand cross-links. The cellular and molecular aspects of the mechanism of action of oxaliplatin have not yet been fully elucidated. However, the intrinsic chemical and steric characteristics of the DACH-platinum adducts appear to contribute to the lack of cross-resistance with cisplatin. To date, mismatch repair and replicative bypass appear to be the processes most likely involved in differentiating the molecular responses to these agents. PMID- 12530523 TI - Cellular and molecular aspects of drugs of the future: meropenem. AB - Meropenem, first synthesized in the late eighties, has become one of the most important beta-lactam antibiotics of the carbapenem subclass used for the treatment of a variety of life-threatening infections. Due to its unique chemical structure, meropenem is not inactivated by the kidney dehydropeptidase I and the majority of microbial beta-lactamases. Its antimicrobial activity is based on its high affinity for the majority of cell wall-synthesizing enzymes, the so-called penicillin-binding proteins, of Gram-positive and -negative bacteria. However, bacteria have evolved several approaches to resist meropenem: (i) by reducing the affinity of the penicillin-binding proteins for the antibiotics, (ii) by decreasing the permeability of the outer membrane of Gram-negative bacteria, (iii) by using efflux pumps, and (iv) by activating zinc-dependent carbapenemases. Meropenem has a low toxicity profile and, in contrast to imipenem, no central nervous system toxicity. PMID- 12530524 TI - Mitochondrial biogenesis in brown adipose tissue is associated with differential expression of transcription regulatory factors. AB - The differentiation of brown adipocytes during late fetal development or in cell culture is associated with enhanced mitochondrial biogenesis and increased gene expression for components of the respiratory chain/oxidative phosphorylation system. We have shown that this is due to a rise in mitochondrial DNA abundance and the corresponding increase in mitochondrial genome transcripts and gene products, as well as to the coordinate induction of nuclear-encoded genes for mitochondrial proteins. We studied how the expression of key components of the transcriptional regulation of mitochondrial biogenesis is regulated during this process. Changes in the expression of nuclear respiratory factor-2/GA-binding protein a and peroxisome proliferator-activated-receptor gamma coactivator-1 (increase) were opposite to those of nuclear respiratory factor-1 and Sp1 (decrease) during the developmental and differentiation-dependent induction of mitochondrial biogenesis in brown fat. These results indicate that the relative roles of transcription factors and coactivators in mediating mitochondrial biogenesis 'in vivo' are highly specific according to the cell type and stimulus that mediate the mitochondriogenic process. PMID- 12530525 TI - Oligo-1,6-glucosidase and neopullulanase enzyme subfamilies from the alpha amylase family defined by the fifth conserved sequence region. AB - The alpha-amylase enzyme family is the largest family of glycoside hydrolases. It contains almost 30 different enzyme specificities covering hydrolases, transferases and isomerases. Some of the enzyme specificities from the family are closely related, others less so. This study, based on the analysis of 79 amino acid sequences, postulates two subfamilies in the framework of the aamylase family: the oligo-1,6-glucosidase subfamily and the neopullulanase subfamily. The specific sequence in the fifth conserved sequence region of the family served as the basis for defining the subfamilies: QpDln for the oligo-1,6-glucosidase subfamily and MPKln for the neopullulanase subfamily. This conserved sequence region is proposed to be the selection marker that enables one to distinguish between the two subfamilies. The 'intermediary' sequence MPDLN can be characteristic of the so-called intermediary group with a mixed enzyme specificity of alpha-amylase, cyclomaltodextrinase and neopullulanase. The evolutionary trees clearly supported the proposed definition of the two subfamilies. PMID- 12530526 TI - Carboxypeptidase E and thrombospondin-1 are differently expressed in subcutaneous and visceral fat of obese subjects. AB - The aim of this study was to identify candidate genes for visceral obesity by screening for genes strongly differentially expressed between human subcutaneous and visceral adipose depots. A cDNA microarray with human adipose-derived cDNAs was used as an initial screening to identify genes that are potentially differentially expressed between human subcutaneous and visceral abdominal fat tissues. For the two best candidates, carboxypeptidase E (CPE) and thrombospondin 1 (THBS1) (EST N72406), real-time RT-PCR was performed to confirm their depot specific expression in extremely obese individuals. Both genes appeared to be strongly differentially expressed, having a higher expression in the visceral depot than in the subcutaneous one. For THBS1, the difference in expression between the depots was greater in women than in men. The involvement of CPE and THBS1 in obesity allows us to suggest that the physiological processes controlled by these genes contribute to depot and gender-related differences in the metabolic complications of obesity. PMID- 12530527 TI - Differential influences of various arsenic compounds on glutathione redox status and antioxidative enzymes in porcine endothelial cells. AB - The cellular response and detoxification mechanisms in porcine endothelial cells (PAECs) to arsenic trioxide (As2O3), sodium arsenite (NaAsO2) and sodium arsenate (Na2HAsO4) were investigated. NaAsO2 at 20 microM for 72 h increased Cu/Zn superoxide dismutase activity resulting in elevated intracellular hydrogen peroxide levels, but As2O3 and Na2HAsO4 did not. Trivalent arsenic compounds increased intracellular oxidized glutathione (GSSG) and total glutathione (GSH) and cellular glutathione peroxidase (cGPX) and glutathione S-transferase (GST) activity, but not glutathione reductase activity. The increased cGPX activity resulted in an elevated cellular GSSG content. Na2HAsO4 increased the cellular GSSG level at 72 h compared to controls. These results imply that the increased GSH content responding to the oxidative stress by trivalent arsenic compounds may be mainly related to the regulation of GSH turnover. The increased GST activity implies that the elevated intracellular GSH level responding to the oxidative stress may be used to conjugate arsenic in PAECs and facilitate arsenic efflux. PMID- 12530528 TI - New endo-beta-1,4-glucanases from the parabasalian symbionts, Pseudotrichonympha grassii and Holomastigotoides mirabile of Coptotermes termites. AB - Abstract. An endo-beta-1,4-glucanase (EG) was purified from the hindgut of an Australian mound-building termite, Coptotermes lacteus. The hindgut extract had a peak separate from those for extracts obtained from the salivary glands and the midgut based on sephacryl S-200 gel chromatography, and also demonstrated an origin different from the endogenous EGs of the termite itself. The recovery was further purified by SDS-PAGE, and its N-terminal amino acid sequence analyzed. This showed high homology to EGs from glycoside hydrolase family (GHF) 7. PCR based cloning methods were applied to the hindgut contents of C. lacteus and individual protozoan symbionts from C formosanus. cDNAs encoding putative EGs homologous to GHF7 members were then identified. The functionality of one of the putative proteins was confirmed by its expression in Escherichia coli. PMID- 12530529 TI - Peptide aptamers: exchange of the thioredoxin-A scaffold by alternative platform proteins and its influence on target protein binding. AB - Peptide aptamers have emerged as powerful new tools for molecular medicine. They can specifically bind to and functionally inactivate a given target molecule under intracellular conditions. Typically, peptide aptamers are generated by screening a randomized peptide expression library, displayed from the Escherichia coli thioredoxin A (TrxA) protein. Here, we transferred peptide moieties from defined TrxA-based peptide aptamers to alternative scaffold proteins, such as the green fluorescent protein and staphylococcal nuclease. Yeast and mammalian two hybrid assays as well as in vitro binding analyses show that the TrxA scaffold can be a major determinant for the binding of peptide aptamers. In addition, we demonstrate that TrxA can correctly display peptide sequences that correspond to the binding domains of natural interaction partners. Therefore, sequence analyses of TrxA-based peptide aptamers, isolated by two-hybrid screening from randomized expression libraries, should also be useful to find cellular binding partners for a given target protein, by homology. PMID- 12530530 TI - Ectoprotein kinase-mediated phosphorylation of FAT/CD36 regulates palmitate uptake by human platelets. AB - Glycoprotein IV (FAT/CD36) has been shown to be phosphorylated by a cAMP dependent, platelet membrane-bound ectokinase. In this study, we demonstrate that ectophosphorylation of FAT/CD36 regulates initial palmitate uptake. This is the first time that short-term regulation of the activity of a long-chain fatty acid carrier could be shown. Phosphorylation of FAT/CD36 was paralleled by a significant decrease in initial palmitate uptake by morphologically and functionally intact platelets. Maximum inhibition of palmitate uptake was achieved at 0.5 nM extracellular ATP, being significantly decreased to 72% compared to the control. Inhibition of palmitate uptake was abolished by co incubation with the specific protein kinase A inhibitor peptide PKI or with beta,gamma-methylene-ATP, and was reversible upon addition of alkaline phosphatase. An extracellular ATP concentration above 5 microM completely prevented the ectophosphorylation-mediated inhibition of palmitate uptake. We conclude that FAT/CD36-mediated palmitate uptake by human platelets is short-term regulated via cAMP-dependent ectophosphorylation of FAT/CD36. PMID- 12530531 TI - Cyclin D3 and p53 mediate sulforaphane-induced cell cycle delay and apoptosis in non-transformed human T lymphocytes. AB - Despite experimental evidence that sulforaphane can exert chemopreventive effects, whether these effects are specific for neoplastic cells is not known. Following our previous demonstration that sulforaphane induces cell cycle arrest and apoptosis in human T lymphoblastoid Jurkat leukemia cells and increases p53 and bax protein expression, we tested sulforaphane on non-transformed phytohemagglutinin-stimulated human lymphocytes. Here, we demonstrate that sulforaphane arrested cell cycle progression in G, phase, through a decrease in the protein expression of cyclin D3. Moreover, sulforaphane induced apoptosis (and also necrosis), mediated by an increase in the expression of p53. These findings suggest that sulforaphane is a growth modulator for T cells. Our in vitro evidence that sulforaphane is active and even cytotoxic in normal as well as transformed lymphocytes raises important questions regarding its suitability for cancer chemoprevention. PMID- 12530532 TI - Pigment epithelium-derived factor (PEDF), a serpin with potent anti-angiogenic and neurite outgrowth-promoting properties. AB - Pigment epithelium-derived factor is a member of the serpin superfamily of proteins, but one that lacks inhibitory properties against either serine or cysteine proteinases. Nevertheless it possesses a number of physiological properties that make it a potentially important protein in regulation of angiogenesis, in neuronal cell survival and in protection of neurons from neurotoxic agents. It is also a protein that is highly up-regulated in the G0 phase of early-passage cells compared with rapidly proliferating cells or senescent cells, and so is also linked to both the cell cycle and cell senescence. The determination of a high resolution X-ray crystal structure of native PEDF provides insight into regions of the protein that may be involved in one or more of these functions. PMID- 12530533 TI - Mechanism of inhibition of RNA polymerase I transcription by DNA-dependent protein kinase. AB - DNA-dependent protein kinase represses RNA polymerase I (Pol I) transcription in vitro. To investigate the mechanism underlying transcriptional repression, we compared Pol I transcription in extracts from cells that either contain or lack the catalytic subunit of DNA-PK (DNA-PKcs). ATP-dependent repression of Pol I transcription was observed in extracts from DNA-PKcs-containing but not deficient cells, required templates with free DNA ends, and was overcome by exogenous SL1, the factor that nucleates initiation complex formation. Order-of addition experiments demonstrate that DNA-PKcs does not inactivate component(s) of the Poll transcription machinery. Instead, phosphorylated Ku protein competes with SL1 for binding to the rDNA promoter and, as a consequence, prevents initiation complex formation. The results reveal a novel mechanism of transcriptional regulation by DNA-PK. Once targeted to DNA, autophosphorylated Ku may displace positive- or negative-acting factors from their target sites, thereby repressing or activating transcription in a gene-specific manner. PMID- 12530534 TI - Evidence for haploinsufficiency of the human HNF1alpha gene revealed by functional characterization of MODY3-associated mutations. AB - Hepatocyte nuclear factor (HNF)1alpha is a homeo-domain-containing transcription factor participating in the regulation of gene expression in liver, kidney, gut and pancreas of vertebrates. In humans mutations in the HNF1 gene are responsible for one form of maturity onset diabetes of the young (MODY3). To define the molecular mechanism underlying MODY3 we investigated the functional properties of seven MODY3-associated mutations representing the spectrum of different kinds of mutations affecting all functional domains of the protein. The mutations introduced into an expression vector encoding human HNF1alpha include in-frame deletion (AN127), nonsense (Q7X, R171X), frameshift (P291fsinsC) and missense (R229Q, P447L, T6201) mutations. Gel retardation and reporter gene assays showed that the functional properties of these mutants differ dramatically, but none of these mutants act in a dominant negative manner. Moreover, the mRNA stability of the mutants AN127, R171X, P291fsinsC and T547E548fsdelTG is impaired compared to the wild-type sequence in transfected cells. This decreased RNA stability is independent of the presence of an intron in the expression vector and thus differs from mechanisms known to be involved in nonsense-mediated decay (NMD). Our results suggest that haploinsufficiency of HNF1alpha is responsible for the pathogenesis of MODY3. PMID- 12530535 TI - In vitro and in vivo stability of the epsilon2zeta2 protein complex of the broad host-range Streptococcus pyogenes pSM19035 addiction system. AB - Streptococcus pyogenes pSM19035-encoded epsilon (10.7 kDa) and zeta (32.4 kDa) proteins are necessary to secure stable plasmid inheritance in bacteria, with zeta acting as toxin that kills plasmid-deprived cells and epsilon as an antitoxin that neutralises the activity of zeta. The epsilon and zeta proteins co purify as a stable complex that, according to analytical ultracentrifugation and gel filtration, exists as epsilon2zeta2 heterotetramer in solution. Co-crystals of the epsilon2zeta2 complex contain epsilon and zeta in 1:1 molar ratio. Unfolding studies monitoring circular dichroic and fluorescence changes show that the zeta protein has a significantly lower thermodynamic stability than the epsilon protein both in free state and in the complex. Proteolytic studies indicate that zeta protein is more stable in the epsilon2zeta2 complex than in the free state. In vivo studies reveal a short half-life of the epsilon antitoxin (-18 min) and a long lifetime of the zeta toxin (>60 min). When transcription translation of a plasmid containing the epsilon and zeta genes was inhibited, cell death was observed after a short lag phase that correlates with the disappearance of the epsilon protein from the background. PMID- 12530536 TI - Effects of antineoplastic agents on cytoplasmic and membrane-bound heat shock protein 70 (Hsp70) levels. AB - Here we report on the study of the effects of different antineoplastic agents, including cytarabine, 4-hydroperoxyifosfamide, the activated form of ifosfamide, vincristine, and paclitaxel, with regard to their capacity to modulate the amount of cytoplasmic and membrane-bound heat shock protein 70 (Hsp70). Hsp70 levels were measured in the myelogenous leukemic cell line K562, in the human colon carcinoma cell line CX2, and in peripheral blood lymphocytes (PBL) under physiological conditions (37 degrees C), and following non-lethal heat shock at 41.8 degrees C. A concentration of 1 microM and an incubation period of 2 h were determined as non-lethal, since none of the different antineoplastic agents induced necrosis or apoptosis in untreated or heat-shocked cells under these conditions. Our results show that tubulin-interacting agents, including vincristine and paclitaxel, but not DNA-interacting agents, including cytarabine and ifosfamide, selectively increase the amount of cytoplasmic Hsp70 in tumor and normal cells, as measured by semi-quantitative Western blot analysis. Mechanistically, a vincristine- and paclitaxel-induced tubulin assembly, as demonstrated by immunofluorescence microscopy, might be responsible for the elevated cytoplasmic Hsp70 levels. Interestingly, an increased membrane expression of Hsp70 following treatment with vincristine or paclitaxel was selectively observed on tumor cells, but not on normal cells. PMID- 12530537 TI - Characterization of the VPS10 domain of SorLA/LR11 as binding site for the neuropeptide HA. AB - The single transmembrane receptor sorLA/LR11 contains binding domains typical for the low-density lipoprotein receptors and a VPS10 domain which, in the related receptor sortilin, binds the neuropeptide neurotensin. SorLA is synthesized as a proreceptor which is processed to the mature form by a furin-like propeptidase. Endogenous sorLA and its hydra homologue HAB bind the neuropeptide head activator (HA). Transiently expressed partial sorLA constructs were investigated for ligand binding. We found that HA binds with nanomolar affinity to the VPS10 domain. The sorLA propeptide also bound to the VPS10 domain, whereas the receptor-associated protein RAP interacted both with the class A repeats and the VPS10 domain. The sorLA propeptide inhibited binding of HA to full-length sorLA and to the VPS10 domain. It also interfered with binding of HA to hydra HAB, which is taken as evidence for a highly conserved tertiary structure of the VPS10 domains of this receptor in hydra and mammals. The propeptide inhibited HA-stimulated mitosis and proliferation in the human neuroendocrine cell line BON and the neuronal precursor cell line NT2. We conclude that sorLA is necessary for HA signaling and function. PMID- 12530538 TI - Substrate specificity and inhibitor studies of a membrane-bound ganglioside sialidase isolated from human brain tissue. AB - A ganglioside-specific sialidase that controls cellular functions such as growth, differentiation, and adhesion has been observed in a variety of cells, but its characterization proved difficult due to firm membrane attachment and lability of the purified enzyme. Here we report on the specificity toward gangliosides and susceptibility to certain inhibitors of a ganglioside sialidase solubilized and purified 5100-fold from human brain. The sialidase removed terminal sialic acids from gangliosides GM3, GM4, GD3, GD2, GD1 a, GD1 b, GT1 b and GQ1 b, but was inactive toward gangliosides with sialic acid in a branching position (as in GM1 and GM2). Lyso-GM3 and -GD1a were good substrates, too, whereas O-acetylation of the sialic acid as in 9-O-acetyl-GD3 caused strongly reduced cleavage. The new influenza virus drug 4-guanidino-2-deoxy-2,3-dehydro-N-acetylneuraminic acid (Zanamivir) exhibited an IC50 value of about 7 x 10(-5) M that was in the range of the 'classical' sialidase inhibitor 2-deoxy-2,3-dehydro-N-acetylneuraminic acid; the bacterial sialidase inhibitor 4-nitrophenyloxamic acid, however, was ineffective. The glycosaminoglycans heparan sulfate, heparin, chondroitin sulfates A and B, as well as dextran sulfate and suramin, were all strongly inhibitory, suggesting that glycosaminoglycans present on the cell surface or in the extracellular matrix may influence the ability of the sialidase to alter the ganglioside composition of the membrane. PMID- 12530539 TI - Specificity determinants in the interaction of apolipoprotein(a) kringles with tetranectin and LDL. AB - Lipoprotein(a) is composed of low density lipoprotein and apolipoprotein(a). Apolipoprotein(a) has evolved from plasminogen and contains 10 different plasminogen kringle 4 homologous domains [KIV(1-110)]. Previous studies indicated that lipoprotein(a) non-covalently binds the N-terminal region of lipoprotein B100 and the plasminogen kringle 4 binding plasma protein tetranectin. In this study recombinant KIV(2), KIV(7) and KIV(10) derived from apolipoprotein(a) were produced in E. coli and the binding to tetranectin and low density lipoprotein was examined. Only KIV(10) bound to tetranectin and binding was similar to that of plasminogen kringle 4 to tetranectin. Only KIV(7) bound to LDL. In order to identify the residues responsible for the difference in specificity between KIV(7) and KIV(10), a number of surface-exposed residues located around the lysine binding clefts were exchanged. Ligand binding analysis of these derivatives showed that Y62, and to a minor extent W32 and E56, of KIV(7) are important for LDL binding to KIV(7), whereas R32 and D56 of KIV(10) are required for tetranectin binding of KIV(10). PMID- 12530540 TI - Amylin-induced cytotoxicity is associated with activation of caspase-3 and MAP kinases. AB - Nanomolar concentrations of human amylin promote death of RINm5F cells in a time- and concentrationdependent manner. Morphological changes of chromatin integrity suggest that cells are predominantly undergoing apoptosis. Human amylin induces significant activation of caspase-3 and strong and sustained phosphorylation of stress-activated protein kinases, c-Jun N-terminal kinase (JNK) and p38, that precedes cell death. Extracellular signal-regulated kinase (ERK) activation was not concomitant with JNK and/or p38 activation. Activation of caspase-3 and mitogen-activated protein kinases (MAPKs) was detected by Western blot analysis. Addition of the MEK1 inhibitor PD 98059 had no effect on amylin-induced apoptosis, suggesting that ERK activation does not play a role in this apoptotic scenario. A correlative inhibition of JNK activation by the immunosuppressive drug FK506, as well as a selective inhibition of p38 MAPK activation by SB 203580, significantly suppressed procaspase-3 processing and the extent of amylin induced cell death. Moreover, simultaneous pretreatment with both FK506 and SB 203580, or with the caspase-3 inhibitor Ac-DEVD-CHO alone, almost completely abolished procaspase-3 processing and cell death. Thus, our results suggest that amylin-induced apoptosis proceeds through sustained activation of JNK and p38 MAPK followed by caspase-3 activation. PMID- 12530541 TI - 2-alkyl-3-Alkylamino-2H-Benzo- and pyridothiadiazine 1,1-dioxides: from K+ATP channel openers to Ca++ channel blockers? AB - A series of 2-alkyl-3-alkylamino-2H-benzo- and 2-alkyl-3-alkylamino-2H-pyrido[4,3 e]-1,2,4-thiadiazine 1,1-dioxides, structurally related to BPDZ 44 and BPDZ 73, two potent pancreatic B-cells K+ATP channel openers, were synthesized and tested on rat pancreatic islets (endocrine tissue) as well as on rat aorta rings (vascular smooth muscle tissue). Alkylation of the 2-position led to double bond tautomerization and formation of compounds with a 2H-conformation. In contrast to the previously described pyridothiadiazine dioxides, such as BPDZ 44, and 7 chlorobenzothiadiazine dioxides, such as BPDZ 73, the 2-alkyl-substituted analogs were found to be poorly active on the insulin releasing process although most drugs exhibited a vasorelaxant activity. As a result, the new 2-alkyl-substituted pyridinic compounds expressed a selectivity profile (vascular smooth muscle tissue vs pancreatic tissue) opposite to that of their non-alkyl-substituted counterparts, i.e. BPDZ 44. Additional investigations revealed that, in contrast to their non 2-alkyl-substituted analogs, the most interesting 2-methyl substituted derivatives did not express the pharmacological profile of classical K+ATP channel openers. The pharmacological results rather suggest that alkylation of the 2-position of the thiadiazine ring led to drugs that could act as Ca2+ channel blockers rather than as potassium channel openers. PMID- 12530542 TI - Pre-loading of chlorophyll synthase with tetraprenyl diphosphate is an obligatory step in chlorophyll biosynthesis. AB - The reaction of recombinant chlorophyll synthase from Avena sativa, expressed in Escherichia coli, was investigated. To verify the identity of the recombinant and native enzymes, reaction rates were determined for both enzyme preparations with several chlorophyllide analogs. The rates of esterification of these modified substrates ranged from 0 to 100% of the rate with the natural substrate, and were nearly identical for both enzyme preparations. The Lineweaver-Burk plot for variation of both chlorophyllide a and phytyl diphosphate concentration showed parallel lines, indicative of a 'ping-pong' mechanism. Pre-incubation with phytyl diphosphate exhibited an initial rapid reaction phase, which did not occur after pre-incubation with chlorophyllide. We conclude that the tetraprenyl diphosphate must bind to the enzyme as the first substrate and esterification occurs when this pre-loaded enzyme meets the second substrate, chlorophyllide. Approximately 10-17% of the recombinant enzyme were pre-loaded with phytyl diphosphate under the experimental conditions. The rapid reaction phase is also observed for the chlorophyll synthase reaction in etiolated barley leaves in addition to the well known slow phase. This indicates that pre-loading of the enzyme with tetraprenyl diphosphate is also the basis for the reaction in vivo. PMID- 12530543 TI - Molecular and immunological characterization of profilin from mugwort pollen. AB - In late summer in Europe, pollen of mugwort is one of the major sources of atopic allergens. No information about the complete molecular structure of any mugwort allergen has been published so far. Here we report the isolation and characterization of mugwort pollen cDNA clones coding for two isoforms of the panallergen profilin. Thirty-six percent of the mugwort-allergic patients tested displayed IgE antibodies against natural and recombinant profilin, and no significant differences were observed in the IgE-binding properties of the isoforms. One profilin isoform was purified to homogeneity and detailed structural analysis indicated that the protein exists in solution as dimers and tetramers stabilized by sulfydryl and/or ionic interactions. Profilin monomers were detectable only after exposure of multimers to harsh denaturing conditions. Dimers and tetramers did not significantly differ in their ability to bind serum IgE from mugwort pollen-allergic patients. However, oligomeric forms might have a higher allergenic potential than monomers because larger molecules would have additional epitopes for IgE-mediated histamine release. Profilin isolated from mugwort pollen also formed multimers. Thus, oligomerization is not an artifact resulting from the recombinant production of the allergen. Inhibition experiments showed extensive IgE cross-reactivity of recombinant mugwort profilin and profilin from various pollen and food extracts. PMID- 12530544 TI - The archaeal respiratory supercomplex SoxM from S. acidocaldarius combines features of quinole and cytochrome c oxidases. AB - The hyperthermoacidophilic archaeon Sulfolobus acidocaldarius has a unique respiratory system with at least two terminal oxidases. Genetic and preliminary biochemical studies suggested the existence of a unique respiratory supercomplex, SoxM. Here we show (i) that all respective genes are translated into polypeptides, and (ii) that the supercomplex can be separated from the alternative oxidase SoxABCD and in that way characterized in a catalytically competent form for the first time. It acts as a quinol oxidase and contains a total of seven metal redox centers. One of it--the blue copper protein sulfocyanin--functionally links two subcomplexes. One is a bb3-type terminal oxidase moiety containing CuA and CuB, whereas the other consists of a Rieske FeS protein and a homolog to cytochrome b--in this case hosting two hemes As. Based on a 1:1 stoichiometry, 1 mol complex contains 6 mol Fe and 4 mol Cu. Its activity is completely inhibited by cyanide and strongly by aurachin-C and -D derivatives as inhibitors of the quinol binding site. These data suggest that the complex provides two proton pumping sites. Interestingly, subunit-II reveals an unusual pH dependence and is proposed to act as a pH sensor as well as a regulator of catalytic activity via a reversible transition between two states of the CuA ligation. This is a novel hint at how S. acidocaldarius can adapt to and survive in its extreme natural environment. PMID- 12530545 TI - Benzodiazepine binding studies on living cells: application of small ligands for fluorescence correlation spectroscopy. AB - We demonstrate the applicability of fluorescence correlation spectroscopy (FCS) for receptor binding studies using low molecular weight ligands on the membranes of living nerve cells. The binding of the benzodiazepine Ro 7-1986/602 (N-des diethyl-fluorazepam), labeled with the fluorophore Alexa 532, to the benzodiazepine receptor was analyzed quantitatively at the membrane of single rat hippocampal neurons. The values obtained for the dissociation constant Kd = (9.9 +/- 1.9) nm and the rate constant for ligand-receptor dissociation kdisS = (1.28 +/- 0.08) x 10(-3) s(-1) show that there is a specific and high affinity interaction between the dye-labeled ligand (Ro-Alexa) and the receptor site. The binding was saturated at approx. 100 nM and displacement of 10 nM Ro-Alexa, with a 1,000-fold excess of midazolam, showed a non-specific binding of 7-10%. Additionally, two populations of the benzodiazepine receptor that differed in their lateral mobility were detected in the membrane of rat neurons. The diffusion coefficients for these two populations [D(bound1) = (1.32 +/- 0.26) microm2/s; D(bound2) = (2.63 +/- 0.63) x 10(-2) microm2/s] are related to binding sites, which shows a mono-exponential decay in a time-dependent dissociation of the ligand-receptor complex. PMID- 12530546 TI - Individual recombinant thyroglobulin type-1 domains are substrates for lysosomal cysteine proteinases. AB - Thyroglobulin contains 11 repeats of a motif called thyroglobulin type-1 domain that show sequence similarity to some proteins exhibiting inhibitory activity against cysteine proteinases. Here we report that thyroglobulin decreases the activity of cathepsins B, H, L, and papain. To examine the possible involvement of particular type-1 domains in that decrease of activity, some individual thyroglobulin type-1 domains were expressed in E. coli. These recombinant domains proved to be substrates for cathepsins B, H, L, and papain instead of inhibitors. The cleavage points with cathepsins B and L on the second and the fourth domains were determined. The possible reasons for degradation are discussed. PMID- 12530547 TI - Characterization of legumain. AB - The mammalian legumain, also called asparaginyl endopeptidase (AEP), is critically involved in the processing of bacterial antigens for MHC class II presentation. In order to investigate the substrate specificity of AEP in the P1' position, we created a peptide library and digested it with purified pig kidney AEP. Digestion was less efficient only when proline was in the P1' position. Maximum AEP activity was found in lysosomal fractions of different types of antigen presenting cells (APC). When the multiple sclerosis-associated autoantigen myelin basic protein (MBP) was digested with AEP, the immunodominant epitope 83-99 was destroyed. Myoglobin as an alternative substrate was AEP resistant. These results suggest an important, but not necessarily critical role for AEP in lysosomal antigen degradation. PMID- 12530548 TI - Molecular cloning and chimerisation of an inhibitory anti-cathepsin B antibody and its expression in Chinese hamster ovary cells. AB - The lysosomal cysteine protease cathepsin B is one of several proteases that have been linked to tumour progression. Its increased expression and secretion in tumour cells may facilitate the degradation of extracellular matrix proteins, leading to tumour cell invasion and metastasis. Specific inhibitory monoclonal antibodies are a possible alternative to synthetic inhibitors as a therapeutic tool for cancer treatment. An inhibitory monoclonal antibody, which binds to an epitope near the active site of cathepsin B and inhibits its proteolytic activity, was prepared and its effect on invasion of ras-transformed MCF-10A neoT cells was tested in vitro. Here we present the nucleotide sequences of the heavy and light chains of the inhibitory antibody and compare them to the murine immunoglobulin germline sequences for possible somatic hypermutations. Since no harmful mutations were found, a mouse/human chimeric antibody was constructed by fusing murine V(H) and V(L) variable regions of the inhibitory antibody with human gamma 1 and K constant regions, respectively. Chinese hamster ovary K1 cells were co-transfected with expression vectors pcD-NA3L and pcDNA3H and the reactivity of the isolated chimeric antibody was tested by ELISA and Western blotting. We could demonstrate an inhibitory effect of the chimeric antibody. PMID- 12530549 TI - Application of peptides containing the cleavage sequence of pro-TNFalpha in assessing TACE activity of whole cells. AB - Tumor necrosis factor-alpha (TNFalpha) is presumably shed from cell membranes by TNFalpha-cleaving enzyme (TACE). The peptides SPLAQAVRSSSR and Dabcyl-LAQAVRSSSR Edans, each encompassing the cleavage sequence of pro-TNFalpha recognized by TACE, were applied to intact umbilical vein endothelium (HUVEC), peripheral blood leukocytes (PBL) and the mast cell line HMC-1, which express TACE, to homogenates of rat heart tissue and to membrane and cytoplasmic extracts of PBL. Formation of SPLAQA (specific cleavage) was determined by HPLC, while cleavage (specific plus non-specific) of Dabcyl-TNFalpha-Edans was followed over time by measuring fluorescence. Participation of TACE was assessed from inhibition due to the drug TAPI-2. Incubation with recombinant human TACE gave specific cleavage, fully inhibitable by TAPI-2 (IC50 < 0.1 microM). HUVEC rapidly degraded TNFalpha peptide, but in a non-specific manner (no SPLAQA detectable) and 50 microM TAPI-2 was without effect. Fluorescence was evoked when Dabcyl-LAQAVRSSSR-Edans was incubated with HMC-1 or PBL and also with cytoplasmic and membrane fractions of lysed PBL, but in no case was there significant inhibition by TAPI-2. However, marginal (10%) inhibition of fluorescence by 50 microM TAPI-2 was observed with homogenized heart tissue. This contained TACE, about 75% of which was without the inhibitory cysteine switch (Western blot). In conclusion, simple peptide analogs of pro-TNFalpha cannot be employed as substrates for measuring membrane TACE activity, largely due to extensive non-specific proteolytic cleavage by whole cells and cell extracts. PMID- 12530551 TI - Effects of duodenal seal oil administration in patients with inflammatory bowel disease. AB - Long-chain n-3 PUFA in fish oil have modulating effects on inflammatory responses. The aim of this open pilot study was to investigate whether duodenal seal oil administration would benefit patients with inflammatory bowel disease (IBD). Seal oil (10 mL) was administered three times a day directly into the distal part of the duodenum via a nasoduodenal feeding tube for 10 d in 10 patients, 5 of whom had Crohn's disease and 5 ulcerative colitis. Nine of the 10 patients suffered from IBD-associated joint pain. Various parameters of disease activity and FA incorporation in tissues were analyzed before and after treatment. Following seal oil therapy, joint pain index, disease activity, and serum cholesterol level were significantly decreased, whereas the n-3 to n-6 ratio both in intestinal biopsies and blood was significantly increased. Measures of calprotectin concentration in gut lavage fluid, intestinal permeability, and lipid peroxidation were not significantly changed. The results suggest positive effects of seal oil in patients with IBD, especially on IBD-associated joint pain. Further controlled studies are warranted. PMID- 12530550 TI - Supplementation with a pine bark extract rich in polyphenols increases plasma antioxidant capacity and alters the plasma lipoprotein profile. AB - Pycnogenol (PYC), an extract of French maritime pine bark (Pinus pinaster), is a potent antioxidant with potential health benefits. Its bioavailabilty has previously been shown by urinary excretion studies of constituents and metabolites of PYC. The aim of this study was to test the effect of PYC supplementation on measures of oxidative stress and the lipid profile in humans. Twenty-five healthy subjects received PYC (150 mg/d) for 6 wk. Fasting blood was collected at baseline, after 3 and 6 wk of supplementation, and again after a 4 wk washout period. After 6 wk of supplementation with PYC, a significant increase in plasma polyphenol levels was detectable, which was reversed after the 4-wk washout phase. The antioxidant effect of PYC was demonstrated by a significant increase in oxygen radical absorbance capacity (ORAC) in plasma throughout the supplementation period (P < 0.05). The ORAC value returned to baseline after the 4-wk washout period. Moreover, in addition to its antioxidant effects, PYC significantly reduced LDL-cholesterol levels and increased HDL-cholesterol levels in plasma of two-thirds of the subjects. While the LDL changes reversed during washout, the HDL increase did not. There was no significant difference in LDL oxidizability or plasma lipid peroxides following PYC supplementation. Hence, following oral supplementation in humans, PYC significantly increases antioxidant capacity of plasma, as determined by ORAC, and exerts favorable effects on the lipid profile. PMID- 12530553 TI - Effect of exercise on FA profiles in n-3 FA-supplemented and -nonsupplemented premenopausal women. AB - The purpose of this double-blind study was to investigate the influence of exercise on the FA profile of the non-esterified FA (NEFA) and phospholipid fractions in plasma of sedentary women supplemented with n-3 FA vs. women supplemented with oil containing no n-3 FA. Twenty sedentary, premenopausal women were randomly assigned to receive 12 capsules daily of either fish oil (3.5 g EPA and 2.4 g DHA per day, each as the ethyl ester) or evening primrose oil capsules (no detectable EPA or DHA). Each subject consumed the capsules for one menstrual cycle. At the end of the supplementation period, the sedentary subjects underwent an acute exercise trial [55% maximal oxygen consumption (VO2 max), 45 min] on a cycle ergometer. Two subjects in the fish oil group were removed from all calculations owing to noncompliance for reasons not related to side effects. There were no changes in the phospholipid composition of either group of women after exercise. In both control and fish oil-supplemented women, NEFA levels in general rose after exercise. There were no changes in the percentage of any given individual NEFA in either supplementation group. However, absolute levels of certain individual NEFA (16:0, 18:0, 18:1, and 18:3n-3) increased with exercise. Women supplemented with fish oil had increased levels of n-3 NEFA IEPA, DHA, and docosapentaenoic acid (DPA)] prior to exercise. Exercise did not, however, increase the absolute levels of n-3 NEFA in the blood. PMID- 12530552 TI - EPA, but not DHA, decreases mean platelet volume in normal subjects. AB - The first indication of platelet activation is an increase in mean platelet volume (MPV). n-3 FA are known to inhibit platelet function and to reduce the risk for coronary heart disease. The purpose of this study was to determine the effects of EPA and DHA on MPV. Healthy subjects received olive oil placebo for 4 wk and then were randomly assigned to receive 4 g of ethyl esters of either safflower oil (n = 11), EPA (n = 10), or DHA (n = 12) for 4 wk. At the end of placebo run-in and treatment periods, MPV (fL; mean +/- SEM) and platelet count (PLT-CT; 10(3)/microL blood) were measured in the basal state and after ex vivo stimulation with collagen (10 microg/mL), cold (4 degrees C), and heat (37 degrees C). Unlike DHA, EPA lowered MPV as compared with safflower oil (7.2 +/- 0.1 vs. 7.5 +/- 0.1 fL; P < 0.05) and raised PLT-CT (211 +/- 18 vs.192 +/- 18 10(3)/microL; P < 0.05) in the fasting state. Collagen and cold significantly increased MPV whereas heat lowered MPV regardless of treatment. All stimuli decreased PLT-CT. EPA significantly increased platelet EPA (0.2 +/- 0.1 vs. 3.3 +/- 0.4%) and docosapentaenoic acid (DPA; 2.2 +/- 0.3 vs. 2.9 +/- 0.3%) concentrations, but not DHA. DHA treatment significantly increased DHA (1.4 +/- 0.2 vs. 4.1 +/- 0.5%) and DPA (2.0 +/- 0.4 vs. 3.0 +/- 0.4%) concentrations, but not EPA. In conclusion, EPA, but not DHA, reduces platelet activation, an early step in platelet aggregation. PMID- 12530555 TI - Trans isomeric octadecenoic acids are related inversely to arachidonic acid and DHA and positively related to mead acid in umbilical vessel wall lipids. AB - Long-chain PUFA play an important role in early human neurodevelopment. Significant inverse correlations were reported between values of trans isomeric and long-chain PUFA in plasma lipids of preterm infants and children aged 1-15 yr as well as in venous cord blood lipids of full-term infants. Here we report FA compositional data of cord blood vessel wall lipids in 308 healthy, full-term infants (gestational age: 39.7 +/- 1.2 wk, birth weight: 3528 +/- 429 g, mean +/- SD). The median (interquartile range) of the sum of 18-carbon trans FA was 0.22 (0.13) % w/w in umbilical artery and 0.16 (0.10) % w/w in umbilical vein lipids. Nonparametric correlation analysis showed significant inverse correlations between the sum of 18-carbon trans FA and both arachidonic acid and DHA in artery (r = -0.38, P < 0.01, and r = -0.20, P < 0.01) and vein (r = -0.36, P < 0.01, and -0.17, P < 0.01) wall lipids. In addition, the sum of 18-carbon trans FA was significantly positively correlated to Mead acid, a general indicator of EFA deficiency, in both artery (r = +0.35, P < 0.01) and vein (r = +0.31, P< 0.01) wall lipids. The present results obtained in a large group of full-term infants suggest that maternal trans FA intake is inversely associated with long-chain PUFA status of the infant at birth. PMID- 12530554 TI - Long-chain PUFA supplementation improves PUFA profile in infants with cholestasis. AB - Long-chain PUFA (LCP) deficiency is a frequent complication in cholestatic infants. We investigated the effects of LCP-supplemented formula on EFA status in infants with cholestasis. Twenty-three infants with cholestasis (biliary atresia after surgery, 8; intrahepatic cholestasis, 15) aged 1.9 to 4.9 mon (median 3.1 mon) were randomized to receive commercial infant formulas either without LCP or with LCP from egg phospholipids for 1 mon. Liver tests, nutrient intakes, and plasma phospholipid FA (%w/w) were determined at baseline and after intervention. At baseline, patients had high serum direct bilirubin levels (5.9 +/- 3.0 mg/dL; mean +/- SD), they were malnourished (body fat mass: 40 +/- 13% of normal) and presented with PUFA deficiency [plasma phospholipid PUFA: 28.43%w/w (26.56-30.53) in patients vs. 37.02%w/w (34.53-39.58) in controls; median (1st-3rd quartile)] with elevated Mead acid and palmitoleic acid. LCP-supplemented (n = 11) and nonsupplemented groups (n = 12) did not differ in age, indicators of liver function, and EFA status at baseline. After the intervention, LCP-supplemented infants had higher levels of arachidonic acid [7.2 (5.9-8.8) vs. 4.2 (3.0-5.3) %w/w; P < 0.001] and DHA [2.8 (2.2-3.2) vs. 1.6 (1.0-2.1) %w/w; P < 0.05], accompanied by increased TBARS concentration: 1.9 (1.4-2.2) vs. 1.3 (1.1-1.6) nmol/mL; P < 0.05]. We concluded that LCP-supplemented formulae improve LCP status of infants with severe cholestasis but may enhance lipid peroxidation. PMID- 12530556 TI - Lipoprotein-apolipoprotein changes in renal transplant recipients. AB - Recent studies have demonstrated that the incidence of cardiovascular events occurring with renal transplantation is higher than that in the general population. Renal transplantation modifies the characteristic dyslipidemia of chronic renal failure. In this study the change in lipoprotein and lipid values of 103 transplant recipients after transplantation was investigated. The aim of our work was to examine the short-term and long-term variations in lipid metabolism. The major lipoprotein fractions (VLDL, LDL, HDL) were separated by preparative ultracentrifugation, and TG and cholesterol concentrations were determined in plasma and lipoprotein fractions. Whole plasma apolipoproteins were determined by a rate immunonephelometric technique. In the pretransplant period the patients displayed the typical picture of uremics. After transplantation the most evident alterations in the lipoprotein profile occurred in our case series after 3 mon. The major finding was a 35% reduction in plasma TG. The modifications in the TG-rich lipoproteins of our transplant recipients persisted throughout the observation period. In the initial 3-mon period, total cholesterol remained steady, whereas LDL-cholesterol and total apolipoprotein B showed a significant increase. No significant changes were found in total and transported TG and cholesterol between the 3-mon and the 6-yr values. The substantial stability of cholesterol levels after transplantation and in subsequent reports, as well as a higher incidence of cardiovascular complications, may suggest that the mechanisms responsible for vessel damage must be sought mainly in the structural and physicochemical alterations of the individual lipoprotein fractions or in other risk factors. PMID- 12530557 TI - Dietary long-chain PUFA in the form of TAG or phospholipids influence lymph lipoprotein size and composition in piglets. AB - Several sources of long-chain PUFA (LCP) are currently available for infant formula supplementation. These oils differ in their FA composition, the chemical form of the FA esters [TAG or phospholipids (PL)], and presence of other lipid components. These differences may affect LCP absorption, distribution, and metabolic fate after ingestion. The purpose of this study was to evaluate the influence of different chemical forms of dietary LCP on the composition of lymph lipoproteins. Eighteen pigs (5 d old) were bottle-fed different diets for 2 wk: a control diet (C), a diet containing LCP as TAG from tuna and fungal oils (TF TAG), or a diet containing LCP as PL from egg yolk (E-PL). We measured lipid and FA composition of lymph, main lymph fractions (TAG or PL), and the particle size of lymph lipoproteins. The average diameter of lymph lipoproteins was significantly lower in the E-PL group compared with the control and TF-TAG groups (C: 3902 +/- 384 A; TF-TAG: 3773 +/- 384 A; E-PL: 2370 +/- 185 A). Arachidonic acid and DHA contents in lymph and lymph-TAG were significantly higher in the TF TAG group compared to the E-PL group (0.50 +/- 0.03 and 0.24 +/- 0.03 g/100 g vs. 0.29 +/- 0.04 and 0.12 +/- 0.03 g/100 g, respectively). The addition to the diet of LCP in the form of TAG or PL affected the size of intestinal lipoproteins and also led to a different distribution of these FA in lymph lipoproteins. PMID- 12530559 TI - Molecular species of PC and PE formed during castor oil biosynthesis. AB - As part of a program to elucidate castor oil biosynthesis, we have identified 36 molecular species of PC and 35 molecular species of PE isolated from castor microsomes after incubations with [14C]-labeled FA. The six [14C]FA studied were ricinoleate, stearate, oleate, linoleate, linolenate, and palmitate, which were the only FA identified in castor microsomal incubations. The incorporation of each of the six FA into PC was better than that into PE. The [14C]FA were incorporated almost exclusively into the sn-2 position of both PC and PE. The incorporation of [14C]stearate and [14C]palmitate into 2-acyl-PC was slower compared to the other four [14C]FA. The incorporation does not show any selectivity for the various lysoPC molecular species. The level of incorporation of [14C]FA in PC was in the order of: oleate > linolenate > palmitate > linoleate > stearate > ricinoleate, and in PE: linoleate > linolenate > oleate > palmitate > stearate > ricinoleate. In general, at the sn-1 position of both PC and PE, linoleate was the most abundant FA, palmitate was the next, and oleate, linolenate, stearate, and ricinoleate were minor FA. The activities of oleoyl-12 hydroxylase, oleoyl-12-desaturase seem unaffected by the FA at the sn-1 position of 2-oleoyl-PC. The FA in the sn-1 position of PC does not significantly affect the activity of phospholipase A2, whereas ricinoleate is preferentially removed from the sn-2 position of PC. The results show that (i) [14C]oleate is most actively incorporated to form 2-oleoyl-PC, the immediate substrate of oleoyl-12 hydroxylase; (ii) 2-ricinoleoyl-PC is formed mostly by the hydroxylation of 2 oleoyl-PC, not from the incorporation of ricinoleate into 2-ricinoleoyl-PC; and (iii) 2-oleoyl-PE is less actively formed than 2-oleoyl-PC. PMID- 12530558 TI - Comparative effect of fenofibrate on hepatic desaturases in wild-type and peroxisome proliferator-activated receptor alpha-deficient mice. AB - In this study is presented the effect of fenofibrate, a prototypical peroxisome proliferator of the fibrate class, on wild-type and peroxisome proliferator activated receptor alpha (PPARalpha)-/- mouse liver FA profile, desaturase mRNA levels, and activities. We established that, following peroxisome proliferator exposure, the hepatic FA profile was greatly modified. These modifications in hepatic FA content required the expression of PPARalpha, as they are suppressed in transgenic mice deficient in this nuclear receptor. Following peroxisome proliferator exposure, delta6- and delta5-desaturase mRNA levels and activities were increased in wild-type but not in PPARalpha-deficient mouse liver. These results suggest the involvement of PPARalpha in the control of hepatic delta6- and delta5-desaturases in mice. Their roles in minimizing long-chain PUFA depletion in the liver during peroxisome proliferator exposure are discussed. PMID- 12530560 TI - Lipase-catalyzed hydrolysis of TG containing acetylenic FA. AB - Hydrolysis of symmetrical acetylenic TG of type AAA [viz., glycerol tri-(4 decynoate), glycerol tri-(6-octadecynoate), glycerol tri-(9-octadecynoate), glycerol tri-(10-undecynoate), and glycerol tri-(13-docosynoate)] in the presence of eight microbial lipases was studied. Novozyme 435 (Candida antarctica), an efficient enzyme for esterification, showed a significant resistance in the hydrolysis of glycerol tri-(9-octadecynoate) and glycerol tri-(13-docosynoate). Hydrolysis of acetylenic TG with Lipolase 100T (Humicola lanuginosa) was rapidly accomplished. Lipase PS-D (Pseudomonas cepacia) showed a fair resistance toward the hydrolysis of glycerol tri-(6-octadecynoate) only, which reflected its ability to recognize the delta6 positional isomer of 18:1. Lipase CCL (Candida cylindracea, syn. C. rugosa) and AY-30 (C. rugosa) were able to catalyze the release of 10-undecynoic acid and 9-octadecynoic acid from the corresponding TG, but less readily the 13-docosynoic acid in the case of glycerol tri-(13 docosynoate). The two lipases CCL and AY-30 were able to distinguish the small difference in structure of fatty acyl moieties in the TG substrate. To confirm this trend, three regioisomers of mixed acetylenic TG of type ABC (containing one each of delta6, delta9, and delta13 acetylenic FA in various positions) were prepared and hydrolyzed with CCL and AY-40. The results reconfirmed the observation that AY-30 and CCL were able to distinguish the slight differences in the molecular structure (position of the acetylenic bond and chain length) of the acyl groups in the TG during the hydrolysis of such TG substrates. PMID- 12530561 TI - Ozonation of PC in ethanol: separation and identification of a novel ethoxyhydroperoxide. AB - The product of the ozonolysis of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine in ethanol-containing solvent was analyzed by chemiluminescence detection-HPLC with on-line electrospray MS, and characterized on the basis of NMR spectroscopy and MS in high-resolution fast atom bombardment mode. The reaction yielded a large amount of a novel ethoxyhydroperoxide compound [1-palmitoyl-2-(9-ethoxy-9 hydroperoxynonanoyl)-sn-glycero-3-phosphcholinel. In addition to a structural analysis, we speculate on the reaction pathway and discuss the possibility of ethoxyhydroperoxide as a potentially reactive ozonized lipid in food and biological materials. PMID- 12530562 TI - Synthesis of 9,9,9-trideutero-1,4-dihydroxynonane mercapturic acid (d3-DHN-MA), a useful internal standard for DHN-MA urinalysis. AB - Racemic 1,4-dihydroxynonane mercapturic acid (DHN-MA) and 9,9,9-trideutero-1,4 dihydroxynonane mercapturic acid (d3-DHN-MA) are synthesized on a 400-mg scale (overall yield approximately 40%) by a two-step sequence involving Michael addition of N-acetyl-L-cysteine to methyl 4-hydroxynon-2(E)-enoate or methyl 9,9,9-trideutero-4-hydroxynon-2 (E)-enoate, followed by reduction of the intermediate adducts with lithium borohydride. The requisite starting methyl esters are obtained, respectively, from heptanal or 7,7,7-trideuteroheptanal and methyl 4-chlorophenylsulfinylacetate via a sulfoxide piperidine and carbonyl reaction described in the literature. The 7,7,7-trideuteroheptanal is easily prepared by classical methods in four steps from 6-bromo-1-hexanol. 13C NMR data indicate that DHN-MA as well as d3-DHN-MA are obtained as mixtures of four diastereomers. Preliminary results show that d3-DHN-MA could be used as an internal standard for mass spectrometric quantification of DHN-MA in human urine. PMID- 12530563 TI - Creating a collaborative partnership between academia and service. AB - This article discusses how the experience of instituting preceptored clinical experiences throughout an undergraduate baccalaureate nursing curriculum resulted in a partnership between academia and service. The collaboration between academia and service built on the strengths of each institution to reach a common goal. Integration of the preceptor clinical model is unique in that implementation occurs in the second semester of a four-semester curriculum. Advantages and disadvantages to the model for students, preceptors, and faculty are presented. Unanticipated benefits to both institutions and ideas to help other programs and facilities develop a collaborative partnership for the education of baccalaureate nursing students are provided. PMID- 12530564 TI - Scholarship revisited: a Collaborative Nursing Education Program's journey. AB - The Collaborative Nursing Program in British Columbia is a nursing education program composed of 10 partners, including five community colleges, four university colleges, and one university. The Board of Accreditation of the Canadian Association of University Schools of Nursing granted a 7-year accreditation to the Collaborative Nursing Program partners in November 2000. During the past several years, the partners have begun to view their collaborative curriculum development, implementation, and evaluation through the lens of scholarship. Using Boyer's model of scholarship, they have begun to evaluate their work of discovery, integration, application, and teaching against criteria in Scholarship Assessed: Evaluation of the Professorate. This article describes the journey the collaborative partners undertook in exploring the meaning of scholarship across the partnership and within each nursing school. PMID- 12530565 TI - Faculty-agency partnering for improved client outcomes. AB - Implications for practice and education include recognition of the value of clinical outcome studies to clinicians and administrators in clinical agencies, as well as to nursing education. This project provides an example of the opportunity for mutually beneficial education and practice partnerships. Faculty, staff, and students demonstrated their ability to use the basic model for outcome studies developed by the United Way of America (1996). Teams of staff, faculty, and students were able to design small scale outcome studies that provided valuable information for clinicians and administrators about evaluating and improving the quality of clinical services, as well as for seeking continued funding. It is anticipated that further benefits may be discovered when faculty work with agencies to target program outcomes that will yield the most powerful effect for these agencies. PMID- 12530566 TI - Interdisciplinary partners: nursing and dental hygiene. PMID- 12530567 TI - Using TQM/CQI processes to guide development of independent and collaborative learning in two levels of baccalaureate nursing students. PMID- 12530568 TI - Worksite "lunch and learn": a collaborative teaching project. AB - Overall, the collaborative teaching project in community health nursing was successful in meeting the needs of students, faculty, the HMO, the employer, and the community. Health education program outcomes and the lessons learned from the processes indicated the collaboration should continue and be expanded. In subsequent academic years, faculty and HMO staff worked to identify additional employer groups and to make this type of learning experience available to a larger number of nursing students. As career paths diverged, this specific collaboration dissolved. However, the opportunities and challenges observed in this collaborative teaching project have been carried forward into new collaborations. PMID- 12530569 TI - Community-driven partnerships: a unique RN-to-BSN clinical experience. PMID- 12530570 TI - Facilitating a collaborative partnership with a homeless shelter. PMID- 12530571 TI - Preceptors' perceptions of clinical performance failure. PMID- 12530572 TI - The spectrum of laryngeal neoplasia: the pathologist's view. AB - Several types of neoplastic change with different prognostic implications typically involve the laryngeal squamous epithelium. The purpose of this review is to examine the spectrum of these changes, as well as their relationship to benign squamous epithelial proliferative states. Since these pathological changes are apt to occur in regions where the epithelial lining is typically squamous, it is important to recognize that the epithelium of the larynx varies from stratified squamous to respiratory-type, depending on the location. The lingual (anterior) surface of the epiglottis is lined by a stratified squamous type, while the laryngeal (posterior) surface is stratified squamous merging into respiratory-type. In the larynx, the supraglottic and infraglottic portions are a respiratory-type, which contrasts with the stratified squamous epithelium of the glottis. This typical distribution does show some degree of variability in those patches of squamous epithelium and is frequently seen within the respiratory-type epithelial regions. The junction between the two epithelial types may be abrupt or separated by a transitional zone. PMID- 12530573 TI - Altered expression of beta-catenin during radiation-induced colonic carcinogenesis. AB - Radiotherapy for malignant pelvic disease is commonly accompanied by treatment induced proctitis, and rarely by colorectal cancer. Translocation of the beta catenin protein, which is a key downstream effector of the Wnt signal transduction pathway, is frequently found in colorectal cancer. Nuclear beta catenin enhances transcriptional activity of the cyclin D1 gene in cancer cells. Here, we evaluate the involvement of the Wnt pathway in radiation-induced colon carcinogenesis with rats (n = 36). Beta-catenin, APC, and cyclin D1 expression profiles were analyzed by immunohistochemistry in radiation-induced chronic colon injury including cancers and ulcerative lesions in rats (n = 12 in treated group, n = 12 in control group). In total, 3 cases of invasive adenocarcinomas were developed in the irradiated portion 50 weeks after a single dose of 36 Gy irradiation. Nuclear translocation of beta-catenin was observed in all radiation induced colon cancers, whereas this protein was also found in the cytoplasm and/or nucleus of 9 cases of non-neoplastic irradiated colonocytes. Nuclear translocation of beta-catenin correlated with loss of APC and gain of cyclin D1 expression, suggesting activation of the Wnt pathway during radiation-induced colorectal carcinogenesis. A single dose of 10 Gy was also given for acute injury (n = 12: 3 each in days 0, 3, 5, and 7, respectively). Beta-catenin expression was distributed in the cytoplasm of degenerating glands at day 3 and 5, and was observed in the cell membrane of those glands with histological normalization at day 7 after irradiation. Because translocation of beta-catenin was found in irradiated-colonic mucosa as well as colon cancer, disruption of beta-catenin expression might be one of the early events in radiation-induced colonic carcinogenesis. PMID- 12530574 TI - Malignant mediastinal germ cell tumor with pure hepatoid differentiation. AB - We describe a germ cell tumor of anterior mediastinal origin, with pure hepatoid differentiation and elevated serum AFP in a 41-year-old man. This is the first report of such a neoplasm analyzed by conventional stains and immunohistochemistry. Hepatocellular differentiation was proved by immunoreactivity with HepPar-1 and alpha-fetoproein (AFP), membranous expression of carcinoembryonic antigen (CEA-poly) in a canalicuar pattern, and focal expression of cytokeratin 19 in abortive ductular structures. Our investigation shows that mediastinal germ cell tumors with hepatoid components typically arise in middle-aged men; they are of pure hepatoid differentiation, as demonstrated here, or exclusively associated with yolk sac structures. PMID- 12530575 TI - Fundic gland polyps (Elster's cysts) of the gastric mucosa. A marker for colorectal epithelial neoplasia? AB - The results of several retrospective studies allow us to conclude that the detection of fundic gland polyps (Elster's cysts) is associated with an increased incidence of colorectal tumors. In this survey, we present the results of a prospective study investigating such a possible relationship. A total of 144 patients were investigated, of whom 80 had fundic gland polyps of the gastric corpus mucosa. Sixty-four of these patients underwent a colonoscopy, whereas the other 64 patients belonged to a sex- and age-matched control group with no fundic gland polyps. In the group with known fundic gland polyps, colorectal neoplasias were found in 29 (45.3%) patients. Eight of these patients had (12.5%) adenocarcinomas, 3 (4.7%) had high-grade intraepithelial neoplasia, and 18 (28.1%) had tubular adenomas. In one patient, a synchronous carcinoma of the rectum and the colon was detected. In comparison, 6 patients of the control group (9.3%) developed tubular adenomas and 9 (14.1%) had hyperplastic polyps of the colonic and rectal mucosa. Our results suggest that it is necessary to conduct a careful diagnostic work-up of the colon in patients with gastric fundic gland polyps. PMID- 12530576 TI - HOPE--a new fixing technique enables preservation and extraction of high molecular weight DNA and RNA of > 20 kb from paraffin-embedded tissues. Hepes Glutamic acid buffer mediated Organic solvent Protection Effect. AB - The growing number of molecular pathologic tools that are currently available require material with good long term preservation of morphology, nucleic acids, and antigenic structures. However, pathologic investigations of tissues done at a molecular level are often hampered by the fixatives in use. We thus endeavored to design a new fixing system, including subsequent paraffin-embedding and sectioning, that makes complete pathologic analyses possible, with special consideration of immunohistochemistry (IHC), in situ hybridization (ISH), and molecular pathology. The optimized HOPE (Hepes-Glutamic acid buffer mediated Organic solvent Protection Effect) fixing technique allows us to preserve and extract high molecular weight DNA and RNA of > 20 kbp suitable for downstream applications, such as PCR and RT-PCR from HOPE-fixed, paraffin-embedded tissues that are up to 5 years old. This technique will most probably lead to new impacts on molecular pathology. PMID- 12530577 TI - Expressions of the CIP/KIP family of CDK inhibitor proteins in primary intestinal large B-cell lymphomas: correlation with clinical outcomes. AB - Immunohistochemistry was performed for p21, p27, p57 and p53 on paraffin-embedded tissue sections from 25 patients who had surgically resected intestinal lymphomas. It was then correlated with the patients' clinical course in an attempt to determine the expression patterns and clinical significance of the CIP/KIP family of cyclin-dependent kinase inhibitors in primary intestinal large B-cell lymphomas. p21 immunostaining was positive in 11 cases (44%) and p27 was positive in 8 cases (32%). All cases were p57-negative. p53 immunostaining was positive in 14 cases (56%) and negative in 11 cases (44%). With respect to the relationship between p21 and p53, seven cases were p53+/p21-, seven cases were p53+/p21+, seven cases were p53-/p2l-, and four cases were p53-/p21+. The expression patterns of p21 and p53 did not influence the patient's clinical outcome. However, p27-positive cases had a much higher percentage of patients sustaining a continuous complete remission state (8/8, 100%) as compared to p27 negative cases (10/17, 59%), although this difference was not statistically significant (p = 0.057). These results suggest that p27 immunoreactivity may be associated with a better clinical outcome. However, further study with larger series are planned to determine the clinical significance of p27 overexpression in primary intestinal large B-cell lymphomas. PMID- 12530578 TI - Infection of murine precision cut lung slices (PCLS) with respiratory syncytial virus (RSV) and chlamydophila pneumoniae using the Krumdieck technique. AB - The Krumdieck technique allows the investigation of the so-called precision cut lung slices (PCLS) with a special microtome. It is thus possible to evaluate morphologic changes over a longer period of time using only a small group of animals. Chlamydophila pneumoniae (Cp) and respiratory syncytial virus (RSV) proved to be important causes of pneumonia, rhinitis and exacerbations of asthma bronchiale, as well as of lower respiratory tract infections in young children. PCLS should be tested for their suitability as an in vitro model for these infections. The PCLS were infected with Cp and RSV over different periods of time. Investigations were carried out by light and transmission electron microscopy (TEM). Furthermore, immunofluorescence (IF) studies with antibodies against bacterial or viral proteins and cell-specific markers were done using confocal laser scanning microscopy (CLSM). Non-infected and infected PCLS showed a well-preserved morphology up to 72 hours. After short infection intervals, typical inclusions of Cp or RSV were detected in vacuoles of different cell types. Infection and cell types could be verified using IF. Cytopathic effects were not prominent. Ciliary beat was detectable up to 96 hours after infection. This in vitro technique offers the possibility of studying mechanisms and effects of bacterial and viral infections on viable tissue complexes. PMID- 12530579 TI - Hyaline cell-rich chondroid syringoma: case report and review of the literature. AB - Hyaline cell-rich chondroid syringoma (HCRCS) is a rare benign cutaneous neoplasm composed of cells with eosinophilic hyaline cytoplasm and plasmacytoid features, the origin of which remains elusive. To the best of our knowledge, only eight cases of this entity have been reported so far, and none of them was submitted to a large panel of myoepithelial markers. We report on a case of a previously healthy 29-year-old male patient who presented with a slowly enlarging flesh colored nodule on the palmar aspect of the tenar region of his left hand, measuring 2 cm in maximum diameter. The nodule was "shelled-out" and submitted to light microscopy, immunohistochemistry, and ultrastructural examination. Histopathologic analysis disclosed a lobulated neoplasm composed of hyaline cells with plasmacytoid features showing ovoid nuclei, with occasional invaginations, finely granular chromatin, and discrete nucleoli; the cytoplasm was deeply eosinophilic with occasional dot-shaped paranuclear hyaline inclusions. On immunohistochemical evaluation, hyaline cells were strongly and diffusely positive for S-100 protein, vimentin, pan (CAM 5.2) and high molecular weight (34betaE12) cytokeratins; these cells were focally positive for GFAP, maspin, neuron-specific enolase, and cytokeratin 14. Alpha-smooth muscle actin, epithelial membrane antigen, carcinoembryonic antigen, collagen IV, Gp100 (HMB 45), and p63 were negative in neoplastic hyaline cells. Ultrastructural analysis disclosed cells with ovoid nuclei showing occasional invaginations and nuclear pockets; the cytoplasm was rich in meshworks of non-bundling intermediate filaments and a variable amount of rough endoplasmic reticulum cisternae. Based on our findings and those previously reported, hyaline cells of HCRCS might posses an aberrant myoepithelial differentiation. Most importantly, pathologists need to be aware of the histologic and immunohistochemical features of HCRCS to avoid a misdiagnosis of highly malignant neoplams, such as malignant melanoma and extra-skeletal myxoid chondrosarcoma. PMID- 12530580 TI - Chronic cholecystitis with bone metaplasia. A case report. AB - Bone metaplasia is a rare phenomenon in the gastrointestinal tract. We present a case of a 58-year-old man who underwent laparoscopic cholecystectomy for symptoms of chronic cholecystitis. Histologic examination of the removed gallbladder revealed intramural bone metaplasia in association with chronic cholecystitis. To the best of our knowledge, such a case has not yet been reported. The clinical significance of this finding remains to be elucidated. PMID- 12530581 TI - Dermatofibrosarcoma protuberans with EMa+ cells. Report of a case suggesting perineurial cell differentiation. AB - A case of dermatofibrosarcoma protuberans (DFSP) with epithelial membrane antigen (EMA)-positive cells is described. The tumor was excised from the left groin of a 28-year-old woman. It showed characteristic histologic features of DFSP with typical diffuse immunohistochemical positivity for CD34. Moreover, scattered neoplastic cells expressed EMA, suggesting perineural cell differentiation. Ultrastructural study found perineurial cell features, such as thin long bipolar cytoplasmic processes, pinocytotic vesicles, fragments of external lamina and/or external lamina-like material, attachment plaques, and desmosome-like junctions. This observation, together with previous immunohistochemical findings of EMA positive cells in a subset of DFSPs, strongly suggests perineurial cell differentiation in these tumors. DFSP should be included in the differential diagnosis of EMA-positive spindle cell lesions of superficial soft tissue and skin. PMID- 12530582 TI - Iodine nutrition and breast feeding. AB - A survey of the databanks Medline and Web of science identified studies dealing with maternal and infant iodine nutrition during breast feeding. The iodine concentration of human milk varies widely due to maternal iodine intake. Mean breast milk iodine concentrations are reported as ranging from 5.4 to 2170 microg/L (median 62 microg/L) in worldwide studies. In the few studies that compared length of lactation, gestation length, and parity number, these factors did not significantly affect milk-iodine concentrations. In studies of maternal iodine deficiency, untreated goiter had no impact on breast milk iodine when compared with controls. Iodine in human milk responds quickly to dietary iodine intake, either supplemented or consumed in natural foods. Easily absorbable iodine from foods, supplemental sources, iodine-based medication or iodine-based antiseptic solutions used during parturition, is taken up by the maternal thyroid and mammary glands through the Na(+)/I(-) symporter system. This transmembrane carrier protein transports iodine against a high concentration gradient. Hormonal iodine in breast milk occurs mainly as T-4, but depending on maternal iodine intake, high concentrations of the inorganic form (iodide) are found. In less developed countries, where natural-food-iodine intake is low, adequate maternal iodine nutritional status depends exclusively on enforcement of food iodination. In industrialized countries, maternal iodine intake has increased as a function of increasing consumption of dairy products. The human infant is sensitive to maternal iodine nutrition during fetal development and later during breast feeding. Environmental factors, not directly related to maternal iodine intake, such as intake of selenium and organochlorine pollutants, can affect thyroid hormone homeostasis in breast-fed infants. In spite of low iodine concentrations found in milk of mothers consuming low-iodine natural foods, long lasting or even life-lasting benefits to the breast-fed infant are demonstrable. PMID- 12530583 TI - Determination of chromium in cerebrospinal fluid using electrothermal atomisation atomic absorption spectrometry. AB - A rapid method to determine the chromium content of cerebrospinal fluid (CSF) samples using electrothermal atomization atomic absorption spectrometry (ETA-AAS) with deuterium-arc background correction is described. The chromium concentration in CSF was evaluated by the standard addition method. Sample dilution (1 + 1) with 0.25% (m/v) Triton X-100 and 4.5% (v/v) HNO3 gave the best combination of sensitivity, reproducibility, and low blank reading compared with dilution using other solvents. Within-batch reproducibility was 3.2% for 20 CSF samples, between batch reproducibility was 4.7%. CSF samples from 43 healthy volunteers collected in a manner designed to avoid contamination yielded chromium concentrations of 14.6 +/- 6.3 ng mL(-1). PMID- 12530584 TI - Trace elements in viral hepatitis. AB - In this study, serum trace elements, including selenium (Se), zinc (Zn), copper (Cu), were determined by using Atomic Absorption Spectrophotometer (SpectrAA 250 Plus Zeeman, Varian, Australia) in sera of patients with viral hepatitis (A, B, C, D, E) cases (n = 102), and statistically compared with the controls (n = 52). In viral hepatitis, Cu levels were found as 3.23 +/- 1.02 mg/L, and this value was significantly higher than the control group (1.13 +/- 0.21) (p < 0.01). Both, Se and Zn levels found to be significantly low in viral hepatitis cases (p < 0.01). While Se level was 81.4 +/- 26.01 microg/L in viral hepatitis (n = 101), it was found to be 166.15 +/- 4.58 microg/L in healthy individuals. Meanwhile, Zn levels were 0.230 +/- 0.081 mg/L and 0.748 +/- 0.392 mg/L in hepatitis cases (n = 101) and the control group, respectively. There was no difference amongst viral hepatitis groups classified in regard with agents and clinical manifestation, such as A, acute hepatitis B, chronic hepatitis B, C, D and E. Previously, it was indicated that absorption disorders in gastrointestinal system, especially in chronic cases, were not main causes of decrease of trace elements by iron and several other parameters in sera of the cases. Therefore, we suggest that decrease in Zn and Se levels and elevation in Cu levels are probably resulted from defence strategies of organism and induced by the hormone-like substances. PMID- 12530585 TI - Reference values of selenium in plasma in population from Barcelona. Comparison with several pathologies. AB - Plasma selenium reference values from healthy donors in the metropolitan area of Barcelona are determined. A random sample from 156 healthy adults (control group) is analysed by using electrothermal atomic absorption spectrometry with Zeeman effect background correction. The relationship between several pathologies and Se content is also evaluated. Se content from 64 samples from subjects with chronic renal failure and 54 from subjects suffering from several malignancies are determined and the results are compared to the reference values. Moreover, Se contents are determined and compared in two groups of children, healthy (19 samples) and children of mothers infected with HIV-1 (16 samples). In the control group, Se plasma concentration ranges between 50 and 145 microg x L(-1) (82.2 +/- 17.5 microg x L(-1)). Significantly lower values are found in the two pathologies studied (malignancy and chronic renal failure), compared to the control group. However, no significant differences in Se content are found between the two groups studied regarding malignancy and chronic renal failure. In children of mothers infected with HIV-1, Se status is significantly lower than that of healthy children. PMID- 12530586 TI - Mechanisms implicated in the effects of boron on wound healing. AB - Recently, we demonstrated that boron modulates the turnover of the extracellular matrix and increases TNFalpha release. In the present study, we used an in vitro test to investigate the direct effect of boron on specific enzymes (elastase, trypsin-like enzymes, collagenase and alkaline phosphatase) implicated in extracellular matrix turnover. Boron decreased the elastase and alkaline phosphatase activity, but had no effect on trypsin and collagenase activities. The effect of boron on the enzyme activities was also tested in fibroblasts considered as an in vivo test. In contrast to the results obtained in vitro, boron enhanced the trypsin-like, collagenase, and cathepsin D activities in fibroblasts. Boron did not modify the generation of free radicals compared to the control and did not seem to act on the intracellular alkaline phosphatase activity, However, as it did enhance phosphorylation, it can be hypothesized that boron may affect living cells via a mediator, which could be TNFalpha whose transduction signal involves a cascade of phosphorylations. PMID- 12530587 TI - The effect of in utero hypoxia on fetal heart and brain trace metals. AB - This study determined the effect of in utero hypoxia on fetal heart and brain pro and antioxidant trace metals. Dunkin-Hartley guinea pigs (50-60 days gestation) were exposed to 1 h hypoxia (7% O2/93% N2) followed by 4 h reoxygenation in room air. Fetal hearts and brains were harvested and analyzed for copper, iron, magnesium and zinc. Fetal brain iron was significantly increased 28% after hypoxia and 35% by 1 h posthypoxia. Fetal brain magnesium demonstrated progressive decreases of 18% by 4 h posthypoxia. No significant effects of hypoxia were observed on heart trace metals. These results indicate that prooxidant metals may be increased and antioxidant metals may be decreased in posthypoxic fetal brain during a time when these tissues may be vulnerable to oxidative injury. PMID- 12530588 TI - Oral cadmium exposure throughout puberty does not inhibit secretion of prolactin, GH and ACTH through dopamine metabolism changes in male rat. AB - This work analyzed possible dopamine-mediated cadmium effects on plasma prolactin, GH and ACTH levels and if these changes were related to metal accumulation. Male rats were treated from day 30 to 60 of life with 50 mg/L of CdCl2 in the drinking water. Cadmium exposure decreased the dopamine (DA) metabolism (DOPAC/DA ratio) in all brain areas studied, and plasma levels of prolactin, GH and ACTH were diminished. The cadmium concentration did not increase nor in hypothalamus nor in the pituitary after the metal exposure. These results suggest that cadmium inhibits the secretion of these pituitary hormones and this inhibitory effect is not mediated by dopamine or the degree of metal accumulation. PMID- 12530589 TI - Silicon and iron levels in tissues of animals treated with a "ferrimagnetic ceramic" with oncotherapeutic potential (anti-tumor) value. AB - The stability in a biological environment of an injectable cement with oncotherapeutic potential--consisting of a glass powder of SiO2 (35.6%), CaO (42.4%), P2O5 (17%), Na2O (5%) and 30% of its weight of Fe3O4 dissolved in (NH4)2HPO4 plus NH4H2PO4--was evaluated referring to the release of silicon and iron. The experimental model was the rat, and organs (liver, kidney, spleen, lung, heart, and brain) of the implanted and control animals were collected for quantification of these elements by electrothermal atomization atomic absorption spectrometry methods. In most of the analysed organs no significant difference in the contents of silicon and iron between the implanted and the control animals was found. PMID- 12530590 TI - Cadmium, copper and zinc in tissues of deceased copper smelter workers. AB - Workers at a copper and lead smelter in northern Sweden have a multifactorial exposure to a number of heavy metals. The concentrations of cadmium, copper and zinc in liver, lung, kidney and brain tissues have been determined by atomic absorption spectrometry in 32 deceased long-term exposed male lead smelter workers, and compared with those of 10 male controls. Furthermore, copper and zinc levels in hair and nails were determined by energy-dispersive X-ray fluorescence. The highest cadmium concentrations among both workers and controls were observed in kidney, followed in order by liver, lung and brain. The levels in kidney, liver and lung were all significantly higher in the workers than in the controls (p < 0.03). Among the workers relatively strong positive correlations (p < 0.03) were observed between cadmium concentrations in liver and lung, liver and kidney, liver and brain, and lung and brain. In the exposed workers a positive correlation was observed between cadmium and zinc concentrations in the kidney (rs = 0.38; p = 0.034). This is probably mainly due to the protein metallothionein, which is stored in the kidney, binding equimolar amounts of these two metals. The highest concentrations of copper were found in hair and nails among both workers and controls, followed in order by liver, brain, kidney and lung. The tissue concentrations of copper in brain, lung and kidney were all significantly higher among the smelter workers than in the controls (p < 0.036). Copper levels in lung and age at time of death were positively correlated among the exposed workers (rs = 0.39; p = 0.029). In the same group, positive correlations between copper and zinc concentrations in kidney (rs = 0.45; p = 0.009) and nails (rs = 0.68; p < 0.001) were also observed, reflecting possible biological interactions between these two metals. Among both workers and controls, the highest zinc concentrations were found in hair, followed in order by nails, liver, kidney, brain and lung. Significantly higher tissue concentrations among the workers as compared with the reference group were noted in kidney, liver and brain (p < 0.033). Neither copper nor zinc concentrations in hair and nails seemed to provide a useful measure of the trace element status of the smelter workers. PMID- 12530591 TI - Review of publications. PMID- 12530592 TI - Brain cancer with induction periods of less than 10 years in young military radar workers. AB - The authors have reported on 5 young patients who had brain tumors that appeared within 10 yr of initial occupational exposures to radar. Four of the patients were less than 30 yr of age when the diagnoses were initially made. Brief induction periods that follow high exposures in individual sentinel patients are a recognized indicator of impending group risk, and these periods call attention to the need for precautionary measures. Similarly, reports of short induction periods for brain cancer on the side of the head in which there has been prior use of cell phones may also indicate increased risk. PMID- 12530593 TI - Chronic effects of workplace noise on blood pressure and heart rate. AB - Environmental noise levels in the United States are increasing, yet there are few studies in which the nonauditory effects of workplace noise are assessed. In the current study, the authors examined chronic effects of noise on blood pressure and heart rate in 374 workers at an automobile plant. Data were collected from subjects prior to the start of their workshift. Participants completed questionnaires about diet, alcohol use, lifestyle, noise annoyance, use of hearing protection, noise exposure outside of the work environment, personal and family health histories, and demographic information. Resting blood pressure, heart rate, and body mass index were obtained. Noise exposure levels were extracted retrospectively from company records for each participant for the past 5 yr. Summary statistics were generated for each variable, and the authors performed bivariate correlations to identify any unadjusted associations. The authors then completed statistical modeling to investigate the effects of noise on blood pressure and heart rate, after they controlled for other variables (e.g., gender, race, age). The authors controlled for confounding variables, after which use of hearing protection in high-noise areas was a significant predictor of a decrease in both systolic and diastolic blood pressures. The results suggested that the reduction of noise exposure by means of engineering controls or by consistent use of hearing protection by workers may positively affect health outcomes. PMID- 12530594 TI - Assessment of health effects in New York City firefighters after exposure to polychlorinated biphenyls (PCBs) and polychlorinated dibenzofurans (PCDFs): the Staten Island Transformer Fire Health Surveillance Project. AB - Following an electrical transformer fire in Staten Island, New York, a health surveillance program was established for 60 New York City firefighters and emergency medical technicians exposed to polychlorinated biphenyls (PCBs) and polychlorinated dibenzofurans (PCDFs). Exposure potential was documented after high levels of PCBs and PCDFs were found on transformer and firefighters' uniforms. Personnel received comprehensive medical examinations, and the results were compared with preexposure values. Serum was analyzed for PCBs, PCDFs, and polychlorinated dibenzo-p-dioxins (PCDDs). Follow-up was conducted 9 mo later. Thirty-two of 58 (55%) firefighters reported initial symptoms, and 3 firefighters required brief medical leave. Pulmonary functions, exercise performance, serum liver functions, and serum lipid profiles were normal or unchanged from preexposure baselines. Serum PCBs averaged 2.92 +/- 1.96 ppb (range = 1.9-11.0 ppb). Five (8%) had serum PCBs that were greater than or equal to 6 ppb. Eight (73%) had a significant decrease (p = .05) in serum PCB level at the time of follow-up. Serum toxic equivalency (TEQ [1998 World Health Organization]) for total PCDDs and PCDFs averaged 39.0 +/- 21.5 (n = 48). Eighteen (38%) had elevated TEQs (i.e., > 40). All firefighters had no short-term heath effects. Modern firefighting uniforms are not meant to replace HAZMAT suits, but these uniforms provide protection from this chemical exposure for most firefighters. PMID- 12530595 TI - Radium in Wisconsin drinking water: an analysis of osteosarcoma risk. AB - Epidemiological studies of radium dial painters have found an association between exposure to high-dose radium and bone cancers. However, only limited data exist on the relationship between low doses of radium, as are found in some drinking water sources, and osteosarcoma. The authors conducted a population-based case control study to examine the association between exposure to radium in drinking water and the occurrence of osteosarcoma. Estimates of radium exposure and covariates used were based on subjects' residential ZIP codes. The median radium level was not significantly associated with osteosarcoma (odds ratio = .98; 95% confidence interval = .93, 1.04; p = .56). The authors found no evidence that radium, at current levels in Wisconsin drinking water, resulted in excess cases of osteosarcoma. PMID- 12530596 TI - Ocular, airway, and dermal symptoms related to building dampness and odors in dwellings. AB - The authors examined the relationship between symptoms of Sick Building Syndrome and reports of building dampness and odors. Two hundred thirty-one multifamily buildings built prior to 1961 in Stockholm, Sweden, contained a total of 4,815 dwellings. The authors selected these buildings for study by stratified random sampling. Occupants answered a postal questionnaire that assessed weekly symptoms, personal factors, population density in each apartment, water leakage in the preceding 5 yr, different types of odors, and signs of high indoor air humidity. The response rate was 77%. Independent information on building characteristics was gathered from the building owners and the central building register in Stockholm. Multiple logistic-regression analysis was applied and odds ratios were calculated, with adjustments for age, gender, current smoking, hay fever, population density, type of ventilation, and ownership of the building. In total, 22% reported at least 1 sign of dampness, and 32% reported odor in the dwelling. Condensation on windows, high air humidity in the bathroom, moldy odor, and water leakage were reported from 6.8%, 8.8%, 5.7%, and 13% of the dwellings, respectively. A combination of odor and signs of high humidity was related to an increased occurrence of all symptoms (odds ratios = 2.2-3.6). Similar findings were observed for a combination of odors and a history of water leakage in the past 5 yr (odds ratios = 1.2-4.4). Symptoms increased with the number of signs of dampness. The study indicated that dampness in dwellings, with emissions of odorous compounds, are associated with an increase in symptoms consistent with Sick Building Syndrome. PMID- 12530597 TI - Short-term effects of air pollution on mortality in nine French cities: a quantitative summary. AB - Between 1990 and 1995, 9 French cities provided data on daily air pollution, total mortality, cardiovascular mortality, and respiratory mortality. Personnel in individual cities performed Poisson regressions, controlling for trends in seasons, calendar effects, influenza epidemics, temperature, and humidity, to assess the short-term effects of air pollution. The authors describe results obtained from the quantitative pooling of these local analyses. When no heterogeneity could be detected, a fixed-effect model was used; otherwise, a random-effect model was used. Significant and positive associations were found between total daily deaths in these cities and the 4 air pollution indicators studied: (1) Black Smoke, (2) sulfur dioxide, (3) nitrogen dioxide, and (4) ozone. A 50-microg/m3 increase in Black Smoke (24 hr), sulfur dioxide (24 hr), nitrogen dioxide (24 hr), or ozone (8 hr) was associated with increases in total mortality of 2.9% (95% confidence interval [CI]) = 1.3, 4.4), 3.6% (95% CI = 2.1, 5.2), 3.8% (95% CI = 2.0, 5.5), and 2.7% (95% CI = 1.3, 4.1), respectively. Similar results were obtained for cardiovascular mortality. Except for sulfur dioxide, positive--but not significant--associations were found with respiratory mortality. The internal consistency among the cities studied, as well as consistency with previously published results, favors a causal interpretation of these associations. PMID- 12530598 TI - Exposure to subcutaneously implanted uranium dioxide impairs bone formation. AB - The introduction of uranium particles into subcutaneous tissue is a risk that affects workers engaged in the extraction, purification, and manufacture of uranium, as well as soldiers who are wounded with uranium shrapnel. The authors evaluated the effect of an internal source of an insoluble form of uranium on bone. Uranium dioxide powder (0.125 gm/kg body weight) was implanted subcutaneously in rats. After 30 days, animals exposed to uranium weighed less than controls. Bone formation activity in endochondral ossification and bone growth were also lower in the experimental animals, as evidenced by histomorphometric and morphometric methods. This is the first study to report bone damage resulting from continuous, nonlethal exposure to an insoluble compound of uranium dioxide over a period of 30 days. PMID- 12530599 TI - Passive smoking, excretion of metabolites, and health effects: results of the Leipzig's Allergy Risk Study (LARS). AB - Over a 5-yr period, the Leipzig's Allergy Risk Study (LARS) investigated the influence of typical indoor-contaminant burdens on the development of allergies and upper respiratory tract infections in allergy-prone children. Typical indoor volatile organic compounds (VOCs) and excretion of certain VOC metabolites in urine were measured in children 3 yr of age. Data analyses were based on parent completed questionnaires, exposure measurements, and medical examinations. Evaluation of passive smoking was of special interest. Generally, residences with a high burden of passive smoking had higher benzene concentrations than residences inhabited by nonsmokers. Obstructive bronchitis was observed more frequently in children exposed to increased concentrations of benzene, as well as toluene, styrene, and m,p-xylene. In addition, atopic symptoms were associated with excretion of certain VOC metabolites. For example, the authors found an association between eczema and exposure to toluene and between eczema and increased excretion of the toluene metabolite S-benzylmercapturic acid. The results suggest that if an association with certain health effects is to be demonstrated, evaluation of external exposures should be supplemented with evaluations of internal exposure. PMID- 12530600 TI - Risk of exposure to hepatitis A virus among day-care workers in Israel: implications for preventive measures. AB - Hepatitis A is an important infectious disease in Israel. The high incidence of hepatitis A, the declining prevalence of antibodies against hepatitis A virus throughout the past decade, and the availability of an active immunization for hepatitis A have engendered the identification of occupations in which workers are at increased risk of acquiring the disease. A prevalence study for hepatitis A antibodies was conducted during the first half of 1998 in a random sample of 37 day-care centers in different parts of Israel. A short interview was conducted, and serum samples were then collected from each worker. The control group comprised a sample of factory workers, blood donors, and women in maternity wards. Ninety percent (402 of 446) of the day-care workers had antibodies to hepatitis A. In a logistic-regression analysis, seropositivity was associated with increased age, minimal education, and lack of military service. When the authors compared a subsample of workers aged 18-40 yr with the control group, seropositivity was associated with the following: birth in Israel, duration of residence in Israel, very minimal education, military service, and employment in a day-care center. These results, together with earlier reports, indicate that day-care workers are at high risk of occupational exposure to hepatitis A. These results may imply that seronegative workers have a 2-fold chance of acquiring hepatitis A. Appropriate preventive measures should be considered, and active immunization of susceptible workers should be undertaken. PMID- 12530601 TI - Integrated pest management and residue levels of dichlorodiphenyltrichloroethane (DDT) and hexachlorocyclohexane (HCH) in water samples from rural areas in Gujarat State, India. AB - The authors identified 2 rural areas for study in Gujarat State, India, on the basis of pesticide use in agriculture and vector-control programs. In rural area 1, pesticides were used in both agriculture and vector-control programs, whereas in rural area 2, they were used in agriculture only. Water samples were collected from both areas, and they were analyzed for total dichlorodiphenyltrichloroethane (DDT) and hexachlorocyclohexane (HCH) residues. The mean residue levels of total DDT in rural areas 1 and 2 were 7.56 ng/l and 4.27 ng/l, respectively. The corresponding values for HCH were 6.96 ng/l and 5.53 ng/l, respectively. Residue levels of both pesticides were significantly lower in rural area 2 than in area 1. PMID- 12530602 TI - Chronic fatigue in a population-based study of Gulf War veterans. AB - Fatigue has been associated with illness in veterans of the Gulf War; however, few studies have confirmed self-reported fatigue by using clinical evaluation, and symptomatic veterans have not been evaluated with established criteria for Chronic Fatigue Syndrome (CFS). The authors describe the frequency and clinical characteristics of CFS in a sample of veterans residing in the northwestern United States. The sample was selected randomly from U.S. Department of Defense databases of troops deployed to southwest Asia during the Gulf War. The selected individuals were invited to participate in a clinical case-control study of unexplained illness. Of 799 survey respondents eligible for clinical evaluation, 178 had fatigue symptoms. Of the 130 veterans who were evaluated clinically, 103 had unexplained fatigue, and 44 veterans met the 1994 U.S. Centers for Disease Control criteria for CFS. In this population, the authors estimated a minimum prevalence of any unexplained fatigue to be 5.1%, and of CFS to be 2.2%. The estimated prevalence was greater among females than among males. Cases were similar to healthy controls, as determined by laboratory tests and physical findings. In comparison to several clinical studies of CFS patients, the authors of this study found a lower proportion of veterans who reported a sudden onset of symptoms (19%) vs. a gradual onset (50%). Although it has previously been suggested that veterans of the Gulf War suffer from higher rates of chronic fatigue than the general population, the study results described herein--on the basis of clinical examination of a population-based sample of veterans-actually indicate that an increased rate may indeed exist. Gulf War veterans with unexplained fatigue should be encouraged to seek treatment so that the impact of these symptoms on overall quality of life can be reduced. PMID- 12530603 TI - Fungi genus and concentration in the air of onion fields and their opportunistic action related to mycotic keratitis. AB - The authors determined fungi concentrations and genera from onion fields located in monsoon and nonmonsoon areas of Taiwan, as well as concentrations from a county housing location, which served as the comparison site. The authors explored the relationship(s) among onion harvesting, monsoon characteristics, airborne fungal levels, and mycotic eye infection. Airborne fungal levels in the onion field in the nonmonsoon area were the highest, followed by decreasing levels in the monsoon area and control sites, regardless of the sampling periods. The geometric mean of airborne fungal concentrations was the highest during onion harvesting. Cladosporium was the most dominant fungus during pre- and postharvest seasons, whereas Aspergillus, Fusraium, Acremonium, Alternaria, Penicillium, Monilia, Mycelia, and Rhizopus were increased significantly during harvesting. Although fungal concentrations in monsoon onion fields were lower than concentrations measured in the nonmonsoon area, the invasion of dangerous fungal pathogens following eye trauma by airborne sharp exotics (e.g., onion flakes) dispersed in high wind may account for the increase in corneal ulcer cases that occur in the monsoon area. PMID- 12530604 TI - Tobacco smoke exposure and serum cotinine in a random sample of adults living in Verona, Italy. AB - In this study, the authors attempted to validate answers to smoking-habit questions contained in the European Community Respiratory Health Survey questionnaire. The respondents were invited to visit the chest clinic at Verona, Italy, and their serum cotinine levels were measured. The authors invited each of 504 subjects to complete a respiratory interview and to give a blood sample for a radioimmunoassay serum cotinine measurement. A total of 375 subjects responded, of whom 129 were smokers (34.4%), 79 were exsmokers (21.1%), and 167 (44.5%) had never smoked. Exposure to environmental tobacco smoke was reported by 216 subjects (57.6% [mean exposure = 3.8 hr/day (+/- 3.4 hr/day standard deviation)]). In smokers, serum cotinine levels were directly related to the number of cigarettes smoked/day. The authors excluded from analysis nonsmokers who had serum cotinine levels that were > or = 14 ng/ml, and the resulting mean values were 1.7 ng/ml (+/- 2.1 ng/ml standard deviation) in nonsmokers unexposed to environmental tobacco smoke and 2.6 ng/ml (+/- 2.6 ng/ml standard deviation) (p < .002) in nonsmokers exposed to environmental tobacco smoke. There was a relationship between serum cotinine levels and hours of exposure to environmental tobacco smoke (R2 = .136, p < .05). Serum cotinine, which is an objective and accepted measure of tobacco exposure, confirmed the validity of the European Community Respiratory Health Survey questionnaire with respect to smoking habits and environmental tobacco smoke exposure. PMID- 12530605 TI - One-year occupational exposure to a cold environment alters lung function. AB - Numerous observations have shown that breathing cold air causes bronchospasm and increases respiratory tract secretions in asthmatic patients and normal individuals. However, few studies have been conducted on the respiratory effects of protracted daily exposures to a cold environment. In this 1-yr study, the authors examined individuals who spent 6 hr a day in cold stores (+3 degrees C to + 10 degrees C) and spent approximately 25% of that time at +3 degrees C. The protocol included a questionnaire about clinical symptoms, with measurements of baseline pulmonary function and airway responses to carbachol and to nasal inhalation of cold air (-5 degrees C). Eleven subjects were examined prior to their first occupational exposure to cold, and again following 6 mo and 12 mo of work in the cold environment. Compared with a control group of 6 subjects engaged at the same time but who did not work in cold stores, 6 of 11 individuals who worked 12 mo in a cold environment experienced increased symptoms of rhinitis, sore throat, and cough. Physiological measurements at 6 mo and 12 mo showed a progressive decrease in forced expiratory volume in 1 sec and a progressive increase in the baseline value of central airway resistance. Forced expiratory flow measured between 25% and 75% of vital capacity had decreased at 6 mo, but showed no further change at 12 mo. A progressive enhancement of bronchial reactivity to carbachol was noted at 6 mo and again at 12 mo, but airway response to nasal breathing of cold air did not vary. The authors concluded that 1 yr of daily exposure to a cold occupational environment elicits a modest--but significant--airflow limitation, accompanied by bronchial hyperresponsiveness, with the effects beginning within 6 mo of exposure. PMID- 12530606 TI - Mercury in saliva and the risk of exceeding limits for sewage in relation to exposure to amalgam fillings. AB - The concentration of total mercury in stimulated saliva was studied in humans with dental amalgam fillings and in 2 nonamalgam groups. The probability of exceeding the limits of mercury permitted in wastewater increased proportionally as the number of amalgam-filled surfaces increased. The mercury limit for sewage is 0.05 mg/l (= 250 nmol/l) effluent, according to the Council of European Communities directive 84/156/EEC. In neither of the nonamalgam groups was this limit exceeded, but 20.5% in the amalgam group exceeded the limit (p < .001). The risk of exceeding the limit increased 2-fold for every 10 additional amalgam filled surfaces (odds ratio = 2.0; 95% confidence interval = 1.3, 3.3). These results demonstrate that humans, especially in populated areas, can be a significant source of mercury pollutants. As a consequence of mercury release, bacteria may acquire mercury resistance, as well as resistance to other antimicrobial agents, thus resulting in failure of antibiotic treatment. PMID- 12530607 TI - Exposure to ethylene glycol monomethyl ether: clinical and cytogenetic findings. AB - Glycol ethers are known reproductive and developmental toxins in laboratory animals, but little is known about their genotoxic effects in humans. In the current article, the authors tested the hypothesis that human in utero exposure to ethylene glycol monomethyl ether (EGME) is associated with the development of specific congenital anomalies and elevated levels of chromosome aberrations. The authors conducted a clinical and cytogenetic evaluation of 41 offspring of 28 females occupationally exposed to EGME for an average duration of 4.6 yr. Six offspring of 5 women who were occupationally exposed to EGME during pregnancy exhibited characteristic dysmorphic features that were not observed in 35 offspring of 23 women who worked in the same facility, but who were not pregnant at the time of exposure. Persistent cytogenetic damage was observed exclusively in all 6 in-utero-exposed offspring, but not in their 12 match non-in-utero exposed controls. The study characterizes EGME as a human teratogen, as indicated by the prevalence of characteristic dysmorphic features and persistent cytogenetic damage in individuals exposed in utero to this chemical. PMID- 12530608 TI - Correlation of urinary 1-hydroxypyrene and 2-naphthol with total suspended particulates in ambient air in municipal middle-school students in Korea. AB - The authors investigated Korean municipal middle school students to ascertain whether urinary 1-hydroxypyrene (1-OHP) and 2-naphthol-markers for polycyclic aromatic hydrocarbon (PAH) exposure-reflect PAHs in ambient air. The authors used the beta-ray absorption method, which is an index of ambient-air PAH exposure, to collect total suspended particulate (TSP) data. The authors measured urinary 1 OHP and 2-naphthol concentrations in 137 nonsmoking students in 4 municipal middle schools within 1 km of ambient air monitoring stations. The median concentrations of urinary 1-OHP and 2-naphthol in the study were 0.09 nmole/mol creatinine and 2.19 micromol/mol creatinine, respectively, and the geometric means were 0.10 nmole/mol creatinine and 2.47 micromol/mol creatinine, respectively. Urinary 1-OHP concentration did not correlate significantly with any TSP index. There were significant correlations between urinary 2-naphthol level and the daily mean TSP level calculated for 2 days before survey, for 1 day before survey, and for the day of survey. These data suggest that urinary 2 naphthol may be a good marker for inhalation exposure to PAHs in ambient air. PMID- 12530610 TI - Pregnancy as delirium. PMID- 12530609 TI - Multiple sclerosis (MS) in an Illinois community. PMID- 12530611 TI - Perinatal HIV transmission: developing country considerations. AB - In many developing countries, because the prevalence of maternal HIV infection is high (more than 30% in some sub-Saharan African countries) and the resources commonly used to prevent transmission in developed countries are generally not available, transmission of HIV from mother to infant is a devastating problem. Countries already experiencing infant mortality rates 10- to 20-fold greater than those in developed countries can expect a doubling of infant and childhood mortality due to HIV. Those infants who escape infection themselves can expect to be orphaned in early childhood. Low-cost antiviral therapy can reduce transmission substantially, but many countries do not have the infrastructure to screen pregnant women for HIV and appropriately treat the mothers and infants. In developing countries, reduction in maternal-child transmission is feasible, but will require substantial additional resources and a well-functioning obstetric care system. PMID- 12530612 TI - Elevated monocyte chemotactic protein-1 in amniotic fluid is a risk factor for pregnancy loss. AB - OBJECTIVE: Pregnancy loss after mid-trimester amniocentesis occurs in 0.5-1% of cases and is frequently attributed to the procedure. Accumulating evidence implicates a pre-existing, but clinically silent, intra-amniotic inflammation in the etiology of adverse pregnancy outcome after mid-trimester amniocentesis. Monocyte chemotactic protein-1 (MCP-1) is a potent chemokine produced by a wide variety of cells during the course of an inflammatory response. This study was designed to assess if the amniotic fluid concentration of this chemokine identifies patients at risk for spontaneous abortion and/or fetal death. METHOD: A retrospective case-control study of women who had a mid-trimester amniocentesis was designed. Cases (n = 10) consisted of patients who had a spontaneous pregnancy loss after the procedure, while the control group (n = 84) consisted of patients who had a normal pregnancy outcome after mid-trimester amniocentesis. MCP-1 was measured by a specific enzyme immunoassay (sensitivity, 18.3 pg/ml). The Kolmogorov-Smirnov test was utilized to assess normal distribution of the data. Logarithmic transformation was applied to achieve normality. Statistical analysis was performed using Student's t test. A receiver operating characteristic (ROC) curve analysis was used to select a cut-off to dichotomize amniotic fluid concentrations of MCP-1. RESULTS: MCP-1 was detectable in all amniotic fluid samples. Patients who had a mid-trimester amniocentesis and a subsequent pregnancy loss had a higher mean amniotic fluid log MCP-1 concentration than those with a normal pregnancy outcome (pregnancy loss, mean 2.95 +/- 0.19 pg/ml vs. normal outcome, mean 2.78 +/- 0.19 pg/ml; p = 0.01). A cut-off of > 765 pg/ml was selected by ROC curve analysis (area under the curve, 0.74; p = 0.01). An amniotic fluid concentration of MCP-1 above this level was strongly associated with pregnancy loss (odds ratio, 7.35; 95% confidence interval, 1.7-31.1), a sensitivity of 70%, and a specificity of 76%. CONCLUSION: A subset of women who had a pregnancy loss after a mid-trimester amniocentesis had higher concentrations of the chemokine MCP-1 than those who had a normal pregnancy outcome. Subclinical intra-amniotic inflammation is a risk factor for pregnancy loss after mid-trimester amniocentesis. This observation may have medicolegal and clinical implications. An elevated MCP-1 concentration in amniotic fluid of patients with a pregnancy loss after a mid-trimester amniocentesis indicates that a pathological condition was present at the time of the procedure. PMID- 12530613 TI - Signs of platelet activation, but not lipid peroxidation, in fetal blood associated with functional and structural umbilicoplacental lesions in pregnancies complicated by impaired glucose metabolism. AB - OBJECTIVES: To study the influence of platelet activation and lipid peroxidation in fetal blood on umbilical vascular prostanoid synthesis and placental morphology in diabetic pregnancy. METHODS: The concentrations of thromboxane A2 (TxA2) and malondialdehyde (MDA) were determined in umbilical cord plasma in 21 women with diabetes mellitus/impaired glucose tolerance (DM/IGT) and ten healthy women. Segments from the umbilical artery and vein were incubated and prostacyclin (PGI2) and TxA2 metabolites were determined. Prostanoid synthesis was stimulated with calcium ionophore at a second incubation. Histological examination was carried out in samples from the umbilical cord, membranes and placental parenchyma. Non-parametric statistical analysis was used, with a two tailed p < 0.05 considered statistically significant. RESULTS: Cord plasma TxA2, but not MDA, was higher among DM/IGT women (p = 0.07). There were indications that cord plasma TxA2, but not MDA, was positively correlated with vascular prostanoid synthesis and synthesis capacity. In the umbilical vein, both the basal and stimulated PGI2 production and the stimulated TxA2 production were lower in the DM/IGT group. Ischemic placental lesions were associated with a high TxA, and a low MDA concentration in cord plasma. CONCLUSIONS: Even in less severe forms of impaired glucose metabolism, disturbances in platelet activation significantly affect both biochemical and morphological vessel wall and tissue functions in the umbilicoplacental unit. This could indicate an abnormal programming of fetal cell functions and designate cases at increased risk of developing cellular and organ damage. PMID- 12530614 TI - Intrapartum sonography to determine fetal head position. AB - OBJECTIVE: To investigate the accuracy of intrapartum transvaginal digital examination in defining the position of the fetal head. PATIENTS AND METHODS: In 496 singleton pregnancies in labor at term, the fetal head position was determined by routine transvaginal digital examination by the attending midwife or obstetrician. Immediately before or after the clinical examination, the fetal head position was determined using transabdominal ultrasound by an appropriately trained sonographer who was not aware of the clinical findings. The digital examination was considered to be correct if the fetal head position was within 45 degrees of the ultrasound finding. The accuracy of the digital examination was examined in relation to maternal characteristics and the progress of labor. RESULTS: The position of the fetal head was determined by ultrasound examination in all 496 cases examined. Digital examination failed to define the fetal head position in 166 (33.5%) cases and, in 330 cases where the position was determined, the findings of the digital and sonographic examinations were in agreement in only 163 (49.4%) cases. The rate of correct identification of the fetal position by digital examination increased with cervical dilatation, from 20.5% at 3-4 cm to 44.2% at 8-10 cm, and was higher if the examination was carried out by an obstetrician than a midwife (50% versus 30%) and if there was absence rather than presence of caput (33% versus 25%). CONCLUSIONS: Routine digital examination during labor fails to identify the correct fetal position in the majority of cases. PMID- 12530615 TI - Ultrastructure of human umbilical vessels in pre-eclampsia. AB - OBJECTIVE: Our objective in this study was to investigate the ultrastructure of endothelial and muscle cells of human umbilical vessels in both normal and pre eclamptic pregnancies. METHODS: Ten umbilical cords from pre-eclamptic (36, 38 and 40 weeks) and four from normal pregnancies (40 weeks) were collected immediately after vaginal deliveries. Umbilical veins and arteries were isolated and fixed in phosphate-buffered 2.5% glutaraldehyde solution (pH 7.2) for 4 h and postfixed with 1% osmium tetroxide at 4 degrees C for 2 h. The sections were embedded in Araldit CY 212. Ultrathin sections were stained with uranyl acetate, examined and photographed. RESULTS: Human umbilical vessel endothelial cells showed ultrastructural changes in pre-eclamptic patients. Weibel-Palade bodies and some organelles such as rough endoplasmic reticulum were found in increased numbers in venous endothelial cells. Accumulations of granular material were detected under the venous endothelium. CONCLUSION: The endothelial and muscle cells of the umbilical vessels from pregnancies complicated by pre-eclampsia showed morphological changes. PMID- 12530616 TI - Fetal magnetic resonance imaging in prenatal diagnosis of central nervous system abnormalities: 3-year experience. AB - OBJECTIVE: To evaluate the comparative merits of ultrasound and fetal magnetic resonance imaging (MRI) in the correct antenatal diagnosis of suspected central nervous system abnormalities. METHODS: A retrospective review of 27 consecutive pregnancies referred for fetal MRI for suspected central nervous system abnormalities between July 1998 and July 2001. Women were referred for the MRI examination when further anatomical and/or pathological clarification of the ultrasound scan findings was needed. Antenatal ultrasound scan and MRI were reviewed in relation to the findings on postpartum investigations. RESULTS: Data were complete for 26 pregnancies. The median gestational age at the time of the ultrasound examination was 26 weeks (95% CI 24 weeks 2 days to 28 weeks 1 day). The median gestational age at the time of magnetic resonance imaging was 27 weeks' gestation (95% Cl 26 weeks 1 day to 29 weeks 2 days). Eight fetuses had associated skeletal, renal and/or cardiac abnormalities previously noted on ultrasound examination. MRI confirmed the ultrasound diagnosis in 15/26 cases (58%). It changed the diagnosis to the correct one in 7/26 (27%) and misdiagnosed four cases (15%). Three of the four cases that were misdiagnosed on MRI occurred in the first 18 months of our 36-month experience. CONCLUSION: Ultrasound remains the primary imaging modality for prenatal diagnosis. Fetal MRI appears to be a useful adjunct to ultrasound to confirm or exclude certain abnormalities; this will consequently help in the counselling of parents and assist in planning further management. However, like any imaging technique, the sensitivity and specificity of the test are likely to improve with experience. PMID- 12530617 TI - Effect of betamethasone on maternal glucose. AB - OBJECTIVE: To determine the effect of single and multiple betamethasone courses on maternal fasting and postprandial glucose values. STUDY DESIGN: A prospective cohort study was performed in women receiving betamethasone at 24-34 weeks' gestation. Fasting and 1-h postprandial capillary glucose values were obtained daily following betamethasone therapy for hospitalized patients. A control group comprised outpatients who underwent weekly fasting and postprandial assessments for 3 weeks. Fasting and 1-h postprandial capillary glucose values were compared between control and betamethasone patients using an unpaired t test. RESULTS: Thirty-five women received a single course of therapy, 19 received multiple courses and 28 served as controls. Mean fasting glucose values for control patients fell within a narrow range of 81.6 +/- 10.3 to 82.2 +/- 6.4 mg/dl for weeks 1-3. Of women receiving betamethasone, 59% of fasting glucose values were greater than 90 mg/dl as compared to 16% of control fasting values (p < 0.00 1, chi2 test). Mean 1-h postprandial values for control women ranged from 107.7 +/- 15.1 to 112.3 +/- 20.0 mg/dl for weeks 1-3. Mean 1-h postprandial glucose values were < or = 140 mg/dl following one, two or three courses of betamethasone therapy. CONCLUSIONS: Betamethasone resulted in an acute increase in fasting glucose following a single course of betamethasone, whereas two or more courses of therapy resulted in a continuous elevation of fasting glucose values. One-hour postprandial values were not clinically abnormal. PMID- 12530618 TI - The association of isolated mild fetal hydronephrosis with postnatal vesicoureteral reflux. AB - OBJECTIVE: We evaluated the incidence of vesicoureteral reflux in fetuses with prenatally detected isolated mild fetal hydronephrosis. METHODS: Fetuses with isolated mild fetal hydronephrosis (defined as a fetal renal pelvis anteroposterior diameter of > or = 4 and < 10 mm before 24 weeks' gestational age) were prospectively evaluated with postnatal renal ultrasound and voiding cystourethrography within the first few weeks after delivery. Infants were evaluated regardless of whether or not renal pelvic dilatation was seen on postnatal ultrasound examination. RESULTS: Forty cases of mild fetal hydronephrosis were identified from the 5,432 patients cared for at our institution from February 1996 to December 1998 (overall incidence: 1/136). Cases involving aneuploidy (n = 1) and inadequate follow-up (n = 5) were excluded from the investigation. One fetus with documented mild hydronephrosis early in gestation had spontaneous resolution and did not undergo postnatal evaluation. Of the remaining 33 infants, 32 underwent postnatal renal ultrasound examination and all had voiding cystourethrography. Vesicoureteral reflux was identified in five (15%) of the neonates. Eighty per cent (four out of five) of these infants were male. Resolution of vesicoureteral reflux occurred in 75% (three out of four) of the infants available for follow-up within 2 years of birth. CONCLUSIONS: Isolated mild fetal hydronephrosis is associated with vesicoureteral reflux on postnatal voiding cystourethrography. PMID- 12530619 TI - The effects of labor on infant mortality among small-for-gestational-age infants in the USA. AB - OBJECTIVE: To determine whether the presence of labor affects infant mortality among small-for-gestational-age (SGA) infants. METHODS: Data were derived from the United States national linked birth/infant death data sets for 1995-97. Singleton SGA live births in cephalic presentation delivered at 24-42 weeks' gestation were included. Mortality rates for SGA infants exposed and unexposed to labor were compared, and relative risks (RR) were derived using multivariable logistic regression models, after adjusting for potential confounding factors. RESULTS: Of 986 405 SGA infants, 87.4% were exposed to labor. Infants exposed to labor at 24-31 weeks had greater risks of dying during the early neonatal period (RR 1.79-1.86). Decreased risks of late and postneonatal death were observed at all gestational ages in the presence of labor. CONCLUSIONS: Exposure to labor is associated with an increased risk of early neonatal death among SGA infants, especially at gestational ages below 32 weeks. Future randomized trials are warranted to determine the optimal obstetric management of these high-risk infants. PMID- 12530620 TI - A newborn with a goiter and thyroid dyshormonogenesis. AB - The report presents a case of a full-term male infant born with an enlarged anterior neck mass, of a healthy woman with normal thyroid function and negative thyroid antibodies. After treatment, his free thyroxine and total triiodothyronine levels normalized. A defect in thyroid hormone synthesis was considered the cause of the hypothyroidism. PMID- 12530621 TI - Prenatal diagnosis of Cantrell's pentalogy with conventional and three dimensional sonography. AB - Omphaloceles and gastroschisis are the most common defects of the fetal anterior abdominal wall. The association of an omphalocele with an anterior thoracic wall defect could result from a variety of congenital syndromes of which Cantrell's pentalogy is the most common. For proper surgical scheduling of the neonate, early diagnosis of each of the components of this syndrome is important. The presence of a congenital intracardiac anomaly is the best predictor of neonatal mortality. We present a case of Cantrell's pentalogy diagnosed prenatally with conventional and three-dimensional sonographic imaging, and confirmed at birth. We discuss this case and the reports in the world literature. PMID- 12530622 TI - Experimental investigation on neural cell survival after dielectrophoretic trapping. AB - Negative dielectrophoretic forces can effectively be used to trap cortical rat neurons. The creation of dielectrophoretic forces requires electric fields of high non-uniformity. High electric field strengths, however, can cause excessive membrane potentials by which cells may unrecoverably be changed or it may lead to cell death. In a previous study it was found that cells trapped at 3 Vtt/14 MHz did not change morphologically as compared to cells that were not exposed to the electric field. This study investigates the viability of fetal cortical rat neurons after being trapped by negative dielectrophoretic forces at frequencies up to 1 MHz. A planar quadrupole micro-electrode structure was used for the creation of a non-uniform electric field. The sinusoidal input signal was varied in amplitude (3 and 5 Vtt) and frequency (10 kHz-1 MHz). The results presented in this paper show that the viability of dielectrophoretically trapped postnatal cortical rat cells was greatly frequency dependent. To preserve viability frequencies above 100 kHz (at 3 Vtt) or 1 MHz (5 Vtt) must be used. PMID- 12530623 TI - The influence of exercise duration at VO2 max on the off-transient pulmonary oxygen uptake phase during high intensity running activity. AB - The purpose of this study was to examine the influence of time run at maximal oxygen uptake (VO2 max) on the off-transient pulmonary oxygen uptake phase after supra-lactate threshold runs. We hypothesised: 1) that among the velocities eliciting VO2 max there is a velocity threshold from which there is a slow component in the VO2-off transient, and 2) that at this velocity the longer the duration of this time at VO2 max (associated with an accumulated oxygen kinetics since VO2 can not overlap VO2 max), the longer is the off-transient phase of oxygen uptake kinetics. Nine long-distance runners performed five maximal tests on a synthetic track (400 m) while breathing through the COSMED K4b2 portable, telemetric metabolic analyser: i) an incremental test which determined VO2 max, the minimal velocity associated with VO2 max (vVO2 max) and the velocity at the lactate threshold (vLT), ii) and in a random order, four supra-lactate threshold runs performed until exhaustion at vLT + 25, 50, 75 and 100% of the difference between vLT and vVO2 max (vdelta25, vdelta50, vdelta75, vdelta100). At vdelta25, vdelta50 (= 91.0 +/- 0.9% vVO2 max) and vdelta75, an asymmetry was found between the VO2 on (double exponential) and off-transient (mono exponential) phases. Only at vdelta75 there was at positive relationship between the time run at VO2 max (%tlimtot) and the VO2 recovery time constant (Z = 1.8, P = 0.05). In conclusion, this study showed that among the velocities eliciting VO2 max, vdelta75 is the velocity at which the longer the duration of the time at VO2 max, the longer is the off-transient phase of oxygen uptake kinetics. It may be possible that at vdelta50 there is not an accumulated oxygen deficit during the plateau of VO2 at VO2 max and that the duration of the time at VO2 max during the exhaustive runs at vdelta100, could be too short to induce an accumulating oxygen deficit affecting the oxygen recovery. PMID- 12530624 TI - Free and phospholipid-bound choline concentrations in serum during pregnancy, after delivery and in newborns. AB - The aims of this study were to determine whether serum free choline and phospholipid-bound choline concentrations change during the pregnancy or after childbirth and to determine if the serum choline concentrations of the mother and newborn are correlated. Serum free and bound choline concentrations were 10.7 +/- 0.5 microM and 2780 +/- 95 microM in control, non-pregnant women, and rose significantly (p < 0.001) to 14.5 +/- 0.6 microM and 3370 +/- 50 microM or to 16.5 +/- 0.7 microM and 3520 +/- 150 microM after 16-20 weeks or 36-40 weeks of pregnancy, respectively. Serum free and phospholipid-bound choline fell by 14-22% (p < 0.05-01) after either vaginal delivery or caesarian section, and remained low (by 15-42%; p < 0.05-0.001) for 12 h and then rose toward the baseline within 24 h. In amniotic fluid, free choline and phospholipid-bound choline concentrations were 22.8 +/- 1.0 and 19.6 +/- 0.8 microM or 24.0 +/- 1.5 and 516 +/- 43 microM at 16-20 weeks of gestational age or at term, respectively. In newborns, serum free choline concentrations were higher (p < 0.001) and phospholipid-bound choline concentrations were lower (p < 0.001) than in their mothers. These results show that serum free choline and phospholipid-bound choline concentrations are elevated during the pregnancy, which may be required for an adequate maternal supply of choline to the fetus. These observations are clinically important to determine the ideal dietary intake of choline during the pregnancy. PMID- 12530625 TI - The role of myoglobin in retarding oxygen depletion in anoxic heart. AB - The present study explores the role of myoglobin (Mb) in retarding the development of anoxia in the perfused working rat heart. We examine this phenomenon by analyzing the behavior and the kinetics of Mb oxygenation and cytochrome aa3 (cytaa3) redoxation. Absorbance changes, measured at wavelength pairs specific to Mb and cytaa3, show parallelism between the Mb oxygenation status and the redox states of cytaa3. Induction of anoxia leads to early and accelerated Mb deoxygenation whereas cytaa3 reduction marks a slight delay and its rate is twice slower than that of Mb. Then, when Mb is desatured above 50%, the cytaa3 reduction becomes accelerated. With the reoxygenated perfusion following the anoxia, the rate of Mb reoxygenation is twice faster than that of the cytaa3 reoxidation. When the oxygen-binding function of Mb, in situ in the heart, is abolished by treatment with sodium nitrite (NaNO2), the redox kinetics of cytaa3 show significant perturbations. Induction of anoxia leads to a precocious and accelerated reduction of cytaa3, compared to the same anoxic heart before the treatment. At reoxygenation, the reoxidation rate of cytaa3 decreases significantly, compared to that before the treatment. Similarly, in the nitrite treated heart, the phosphocreatine (PCr) level decreases to 60% of the control, whereas the inorganic phosphate (Pi) level increases to 300%. ATP concentration, however, remains constant. We conclude from these results that Mb may support mitochondrial respiration at the critical levels of the myocardial O2 supply. PMID- 12530626 TI - The human MC2-R gene expression: different aspects of its control. AB - Expression of the adrenocorticotropin receptor (MC2-R) is restricted to adrenocortical cells and is up-regulated by both adrenocorticotropin and angiotensin II through the activation of protein kinase A and protein kinase C pathways, respectively. After cloning of the promoter region of the human MC2-R gene (hMC2-R), we have shown that cyclic AMP-induced regulation of transcriptional activity of the gene is achieved through two SF1 binding elements located in the proximal promoter. On the other hand, regulation by angiotensin II partly involved two AP1 binding sites. Using different primary cell cultures, we have also been able to delineate a region inside the promoter which is responsible for part of the tissue-specific expression of the gene. PMID- 12530627 TI - Agonist activated adrenocorticotropin receptor internalizes via a clathrin mediated G protein receptor kinase dependent mechanism. AB - The physiological effects of the pituitary hormone, adrenocorticotropic hormone (ACTH) on the adrenal are mediated by the melanocortin 2 receptor (MC2R), a G protein coupled receptor (GPCR) that signals via adenylate cyclase to elevate intracellular cyclic AMP (cAMP) levels. The function and expression of the receptor is likely to be a major determinant of the response to ACTH. Following repeated stimulation, the cAMP signal is diminished or desensitized. Prolonged desensitization may involve internalization of the receptor. Internalization may occur by at least two mechanisms--receptor mediated endocytosis via clathrin coated pits and by caveolae mediated internalization. The mode of internalization for the endogenous MC2R in Y1 cells was determined using radiolabelled ACTH. Treatment of Y1 cells with hypertonic sucrose or with concanavalin A, which inhibit clathrin-mediated endocytosis, blocked internalization. Filipin and nystatin, which inhibit caveolae formation, did not influence internalization. A dominant negative GRK2 inhibited internalization whilst the protein kinase A (PKA) consensus site mutant MC2R (S208A) internalized normally. However, dominant negative V53D beta-arrestin-1 did not inhibit ACTH internalization in Y1 cells. In conclusion, it appears that the MC2R in Y1 cells internalizes by a G protein coupled receptor kinase (GRK) dependent clathrin-coated pit mechanism. PMID- 12530628 TI - Roles of several domains identified in the primary structure of salt-inducible kinase (SIK). AB - Salt-inducible kinase (SIK), a 776 amino acids-protein, contains a kinase domain in the NH2-terminal 278 amino acid residues, and the biological functions of its COOH-terminal half have yet to be clarified. Here we describe the roles played by several domains in the SIK molecule. K56, an amino acid residue found in a region similar to the ATP-binding loop of other protein kinases, was essential for carrying out the SIKs phosphorylation reaction. An SNF-1 homology domain (SNH), identified at a peptide stretch from the 317th to the 346th residues, and conserved among all the sucrose-nonfermenting-1 protein kinase (SNF-1) family protein kinases, was important to maintain the SIKs protein conformation in the cells. S577, an amino acid residue found in one of three consensus PKA-dependent phosphorylation motifs, was indeed phosphorylated by PKA. The phosphorylated SIK was found to move out of the nucleus to the cytoplasm. PMID- 12530629 TI - The steroidogenic acute regulatory protein, StAR, works only at the outer mitochondrial membrane. AB - The steroidogenic acute regulatory protein (StAR) facilitates the movement of cholesterol into mitochondria to initiate steroidogenesis, but its site of action on the mitochondria has been uncertain. One model states that StAR has a fairly rigid structure and functions in the intramembranous space (IMS) where it transports cholesterol from the outer mitochondrial membrane (OMM) to the inner mitochondrial membrane (IMM); another model states that StAR works solely on or in the OMM and undergoes a partially open molten globule conformation while picking up and discharging cholesterol. We designed, built and tested a series of StAR fusion proteins designed to immobilize StAR on the OMM, the IMS, or the matrix side of the IMM. Only the constructs at the OMM were active, either in vivo or in vitro. As these data indicated that StAR acts at or in the OMM we hypothesized that StAR' s activity would be proportional to the amount of time it spends on the OMM. To test this hypothesis, we built a series of StAR proteins with altered mitochondrial leaders designed to speed or slow StAR's mitochondrial entry. Cell transfections showed that the constructs that slowed entry had more activity and those designed to speed entry had less activity. Analysis of import kinetics in vitro confirmed that these constructs accelerated import inversely proportional to their activity. These data show that StAR works only on the OMM, providing an unusual example of a protein that exerts its biological activity in a cellular location it occupies only transiently, rather than in the location (the matrix) to which it is targeted. PMID- 12530630 TI - Identification of a receptor for N-POMC peptides. AB - The adrenal cortex is a dynamic organ in which the cells of the outer cortex continually divide. It is well known that this cellular proliferation is dependent on constant stimulation from peptides derived from the ACTH precursor proopiomelanocortin (POMC) since disruption of pituitary corticotroph function results in rapid atrophy of the gland. Although ACTH has often been assumed to be the adrenal mitogen, results from our laboratory suggest that the true mitogen is a fragment derived from the N-terminal of POMC that does not contain the gamma MSH sequence. Since these peptides are not generated during the processing of POMC in the pituitary it has been proposed that the mitogen is generated from circulating pro-gamma-MSH by an adrenal protease. We have recently substantiated this hypothesis by characterizing a serine protease expressed by the adrenal necessary for growth and lead us to propose that N-POMC (1-52) is the adrenal mitogen. Using N-POMC (1-28) linked to a solid support we have extended these studies in an attempt to identify the receptor through which this peptide elicits its actions. Using this approach we have isolated a 80 KDa candidate protein from membranes prepared from the adrenal cortical Y1 cell line. PMID- 12530631 TI - Identification of the nuclear localization domain of salt-inducible kinase. AB - Salt-inducible kinase (SIK), a novel serine/threonine protein kinase from adrenal glands of rats fed with a high-salt diet, is induced by ACTH in Y1 mouse adrenocortical tumor cells. Overexpression of SIK repressed ACTH-mediated expression of CYP11A- and Steroidogenic acute regulatory (StAR)-genes by inhibiting CREB bound to their promoters. Immunocytochemical and GFP fluorocytochemical analyses indicated that SIK was present both in the nucleus and cytosol of resting cells. Responding to ACTH, the nuclear SIK moved to the cytosol. The level of phosphorylation at Ser577, a canonical PKA-phosphorylation site, was elevated by ACTH treatment. The disruption of the serine residue inhibited the nuclear export and enhanced the transcription repression activity of SIK. Various deletion mutants suggested a functional nuclear localization signal was present near Ser577. We conclude that the nucleocytoplasmic shuttling of SIK may play an important role in the transcriptional regulation of the cAMP responsive element (CRE)-dependent gene expression. PMID- 12530632 TI - Protein serine/threonine phosphatase 2A activity is inhibited by cAMP in MA-10 cells. AB - PP1 and PP2A are members of the protein serine/threonine phosphatases (PPs) family and their activities have been proposed as a requirement for hormone- and cAMP-regulated steroid synthesis. These findings raise the question whether the PPs activity is increased by hormonal action in steroidogenic systems. Thus, the aim of the study was to evaluate the action of cAMP on the activity of PP1 and PP2A in MA-10 Leydig cells. Our results demonstrate that 8Br-cAMP stimulation produces a transient inhibition of PP2A activity. In contrast, PP1 activity remains unchangeable. As reported in other steroidogenic cells, cAMP-induced steroidogenesis in MA-10 cells is reduced by Cantharidin (Can) and also by Calyculin A (CA), two chemically unrelated PP1/PP2A inhibitors (data not shown). Taking into account the inhibitory effect of cAMP treatment on PP2A activity, the latest findings result paradoxical. Therefore, we next evaluated the action of these compounds on total protein synthesis. Can 10(-5) M and CA 10(-7) M markedly reduced total protein synthesis (35 and 50% respectively) in MA-10 cells, measured by 35S-methonine incorporation. These results suggest that hormone dependent steroidogenesis is working through inhibition of PP2A-dependent dephosphorylation and the effect of PP1/PP2A inhibitors on steroidogenesis may be due to a general inhibition of protein synthesis rather than to a specific action on StAR protein induction. PMID- 12530633 TI - Oxidized products of linoleic acid stimulate adrenal steroidogenesis. AB - Adrenal steroidogenesis is under complex control, and clinical observations suggest that not all regulators have been identified. We postulated that fatty acid oxidation products found in the diet or formed in the body could affect steroidogenesis. Linoleic acid is a prominent constituent of animal fat and is readily oxidized. We found that several products of linoleic acid oxidation affect production of aldosterone and corticosterone by isolated cells from rat adrenals. We characterized one linoleic acid derivative by gas chromatography/mass spectrometry. It is 12,13-epoxy-9-oxo-10(trans)-octadecenoic acid ("EKODE"). At concentrations between 1 and 30 microM, EKODE stimulated production of aldosterone by zona glomerulosa cells, but at concentrations above 50 microM, it was inhibitory. In zona fasciculata cells, EKODE stimulated corticosterone production at concentrations of 5 microM or greater, and there was no evidence of inhibition at high concentrations. Stimulation of steroidogenesis was observed after 15 min of incubation and continued for at least 2 hrs. The potential relevance of our findings to the hypertension of obesity is discussed. PMID- 12530634 TI - ACTH regulation of mitochondrial acyl-CoA thioesterase activity in Y1 adrenocortical tumour cells. AB - We have previously purified and cloned a phosphoprotein, Arachidonic acid-Related Thioesterase Involved in Steroidogenesis (ARTISt), involved in steroid synthesis through Arachidonic Acid (AA) release. Arachidonic acid-related thioesterase involved in steroidogenesis resulted to be a member of a new family of acyl-CoA thioesterases. The protein was identified by its biocapacity to increase mitochondrial steroidogenesis in a cell free bioassay. In the present study we measure the activity of ARTISt using arachidonoyl-CoA (AA-CoA) as substrate. We demonstrate that ACTH significantly stimulates endogenous mitochondrial thioesterase activity as early as 5 min after ACTH stimulation of Y1 cells. Nordihydroguaiaretic acid (NDGA), an inhibitor of AA release known to affect steroidogenesis, affects the in vitro activity of recombinant ARTISt and also the endogenous mitochondrial acyl-CoA thioesterases. ACTH activation of the enzyme protected ARTISt to the inhibitory effect of NDGA. These results show that an enzyme that release AA from AA-CoA can be regulated in intact cells by steroidogenic hormones. PMID- 12530635 TI - Identification of two alternate splice variants of a novel serine protease expressed in steroidogenic tissues. AB - During the search for the serine protease that cleaves pro-gamma-melatropin to stimulate adrenal growth, we identified another novel protease, which we called Adrenal mitochondrial protease (AmP). In situ hybridisation detected AmP transcripts in steroidogenic tissues such as the brain, testis, in ovarian follicles as well as in the adrenal cortex. Full length cloning identified two splice variants differing by a 222 nucleotide insertion in the 5' end of the short variant. The shorter variant codes for a 371 amino acid protein of 40.7 kDa and computer analysis predicts it to be targeted to the cytosol while the longer 445 amino acid protein of 48.4 kDa is mitochondrial. Cellular targeting was confirmed by tagging with GFP. The short variant was clearly cytosolic however, the cells expressing AmP-Long had large vacuoles, possibly as a result of distended (apoptotic?) mitochondria. Due to the mitochondrial localisation of the long variant of the protease and its expression in steroidogenic tissues, it may be expected to be involved in the steroidogenic pathway, possibly by cleaving steroidogenic acute regulatory protein (StAR). We investigated this by co transfecting AmP-Long with StAR and F2 plasmid into COS-1 cells and measuring the effect on pregnenolone production. It was found that AmP-Long has no effect on steroidogenesis nor cleaves StAR as was shown by western blot analysis using StAR antibody. PMID- 12530636 TI - Modulation of steroid hydroxylase activity in stably transfected V79MZh11B1 and V79MZh11B2 cells by PKC and PKD inhibitors. AB - We recently observed that treatment of CYP11B2-expressing COS-1 cells with the broad range kinase inhibitor, staurosporine (STS), strongly inhibited aldosterone biosynthesis, indicating that the activity of a kinase might be a prerequisite for steroid hydroxylase activity. In an attempt to identify such kinases, we measured conversion of 11-deoxycortisol (RSS) and 11-deoxycorticosterone (DOC) by V79MZh11B1 and V79MZh11B2 cells, respectively, in the presence of STS and also after treatment with the kinase inhibitors chelerythrine, rottlerin and Go 6976. The conversion of both substrates by both cell lines was affected in a selective manner by the kinase inhibitors, suggesting that the activity of the novel PKC delta and either of conventional PKCs or of PKD alter steroid hydroxylation activity, with their influence depending on both the cytochrome P450 tested and on its steroid substrate. PMID- 12530637 TI - Bacterial lipopolysaccharide directly stimulates cortisol secretion in human adrenal cells. AB - Adrenomedullin and pro-adrenomedullin N-terminal 20 peptide (PAMP) are expressed in vascular cells and in the adrenal cortex and medulla. Lipopolysaccharide (LPS), a bacterial product that induces septic shock, is a potent stimulant of adrenomedullin secretion in vascular cell types and is also known to stimulate the hypothalamo-pituitary-adrenal axis (HPA). The present study was designed to investigate the actions of LPS on the human adrenocortical cell line, H295R. Exposure of cells to LPS for 24 hours had no effect on adrenomedullin or PAMP secretion, but was found to significantly and selectively increase cortisol secretion with no effect on aldosterone. Dibutyryl cAMP, however, caused a significant increase in both adrenomedullin and PAMP release over this time period. There are two conclusions which can be drawn from these observations. First that adrenomedullin and PAMP are regulated by different mechanisms in vascular and adrenal cells and second, that LPS is able to directly stimulate cortisol secretion, with implications for the physiological response to septic shock. PMID- 12530638 TI - Tumour-derived human adrenocortical cells express beta-adrenergic receptors: steroidogenic effects of beta-adrenergic input. AB - It is well established that catecholamines have potent actions on adrenocortical function and steroidogenesis in different species. The effect of these substances on steroid production of the human adrenal cell line H295R is the subject of this study. H295R cells were cultured in the presence of the synthetic catecholamine, isoproterenol for four hours. Aldosterone, cortisol, and DHEA secretion was measured using direct radioimmunoassays. Administration of 10(-11)-10(-7) mol/L isoproterenol produced a dose-dependent increase in secretion of aldosterone, cortisol, and DHEA by H295R cells resulting in 3-fold, 2.5-fold, and 2-fold stimulation respectively, relative to basal values. Analysis of mRNA using nested PCR revealed the presence of all three types of beta-adrenergic receptors namely beta1, beta2, and beta3 in H295R cells. Isoproterenol had no effect on the proliferation rate of H295R cells as determined by 3H-incorporation assay and the colorimetric WST-1 cell proliferation assay. PMID- 12530639 TI - The life cycle of the steroidogenic acute regulatory (StAR) protein: from transcription through proteolysis. AB - The Steroidogenic Acute Regulatory (StAR) protein is a mitochondrial protein required for the transport of cholesterol substrate to the P450scc enzyme located in the inner mitochondrial membranes of steroid producing cells. This study suggests that the acute regulation of the rodent StAR gene in the ovary is mediated by two factors, C/EBPbeta and GATA-4. Once translated, the StAR precursor protein is either imported into the mitochondria, or it is rapidly degraded in the cytosol. We predicted that in order to perpetuate StAR activity cycles, imported StAR should turn over rapidly to avoid a potentially harmful accumulation of the protein in sub-mitochondrial compartments. Pulse-chase experiments in metabolically labeled cells showed that: (a) the turnover rate of mature mitochondrial StAR protein (30 kDa) is much faster (t(1/2) = 4-5 h) than that of other mitochondrial proteins; (b) dissipation of the inner membrane potential (-delta psi) by carbonyl cyanide m-chlorophenylhydrazone (mCCCP) accelerates the mitochondrial degradation of StAR; (c) unexpectedly, the mitochondrial degradation of StAR is inhibited by MG132 and lactacystin, but not by epoxomicin. Furthermore, StAR degradation becomes inhibitor-resistant two hours after import. Therefore, these studies suggest a bi-phasic route of StAR turnover in the mitochondria. Shortly after import, StAR is degraded by inhibitor sensitive protease(s) (phase I), whereas at later times, StAR turnover proceeds to completion through an MG132-resistant proteolytic activity (phase II). Collectively, this study defines StAR as a unique protein that can authentically be used to probe multiple proteolytic activities in mammalian mitochondria. PMID- 12530640 TI - Regulation of steroid hormone biosynthesis in R2C and MA-10 Leydig tumor cells: role of the cholesterol transfer proteins StAR and PBR. AB - The MA-10 mouse Leydig tumor cell line produces large amounts of steroids only in response to hormonal stimulation while the R2C rat Leydig tumor cell line is constitutively steroidogenic in nature. In an effort to uncover the potential reasons for constitutive steroidogenesis in R2C cells, we have recently shown that compared to MA-10 cells, R2C cells express much higher levels of the Scavenger Receptor Class B type 1 which results in a higher capacity for cholesteryl ester uptake through the selective uptake pathway. We also found an enhanced expression of Hormone Sensitive Lipase and the Steroidogenic Acute Regulatory protein in these cells and reasoned that they may further facilitate the conversion of cholesteryl esters to free cholesterol and its mobilization to the inner mitochondrial membrane, thus rendering them constitutively steroidogenic. Given the proposed role of the peripheral-type benzodiazepine receptor (PBR) in conferring a constitutively steroidogenic phenotype to the R2C cells, the current study was conducted to investigate the relationship between its expression in MA-10 and R2C cells and correlate it with the constitutive nature of R2C cell steroidogenesis. Our studies show that PBR expression as measured by PK 11195 ligand binding and Western analysis is much higher in MA-10 cells than R2C cells. We also determined that the affinity of ligand binding to the PBR is comparable in the two cell lines, suggesting that PBR is unlikely to be solely responsible for the constitutive nature of R2C cell steroidogenesis. PMID- 12530641 TI - PBR, StAR, and PKA: partners in cholesterol transport in steroidogenic cells. AB - Acute stimulation of cholesterol transport into mitochondria involves the cAMP dependent protein kinase (PKA), peripheral-type benzodiazepine receptor (PBR), and the steroidogenesis acute regulatory (StAR) proteins. We investigated the respective role of these proteins in hormone-induced steroidogenesis. Oligonucleotides antisense, but not sense, to PBR and StAR reduced their respective levels in steroidogenic cells and inhibited hormone-stimulated steroid formation in MA-10 mouse Leydig tumor cells. In search of the proteins regulating PBR we identified a protein, PAP7, which interacts with PBR and the PKA regulatory subunit RIalpha, is present in adrenal and gonadal cells and is found in mitochondria. Overexpression of the full length PAP7 increased the hormone induced steroid production. However, inhibition of PAP7 expression reduced the gonadotropin-induced steroid formation. In search of a PBR functional antagonist that would facilitate the studies on the biological function of PBR, we screened a phage display library. A 7-mer competitive PBR peptide antagonist was identified, which when transduced into Leydig cells inhibited the benzodiazepine and hormone-stimulated steroid production suggesting that the endogenous PBR agonist/receptor interaction is critical for the hormone-dependent steroidogenesis. These data indicate that hormone-induced cholesterol transport and the subsequent steroid formation is a dynamic multistep process involving protein-protein interactions. PMID- 12530642 TI - The role and regulation of the Niemann-Pick C1 gene in adrenal steroidogenesis. AB - The Niemann-Pick C-1 (NPC-1) gene codes for an intracellular membrane glycoprotein that processes low density lipoprotein (LDL)-imported cholesterol. Its absence is characterized by pathologic accumulation of cholesterol in endosomes and lysosomes. Niemann-Pick C-1 is involved in steroidogenesis in the adrenal, ovary, testis, and placenta. We investigated adrenal morphology of the BALB/c NPC-1(-/-) mutant mouse. Adrenals of mutant and wild type mice were similar at three weeks, but were substantially smaller in NPC-1(-/-) mice at eight weeks. The major histological difference was reduction in the thickness of the zona fasciculata, due to decreased cell volume. Lipid accumulation, revealed by Oil Red-O staining, was restricted to the zona fasciculata in wild type mice, but was found throughout the cortex in NPC-1 mutants. The 5'-flanking region of the mouse NPC-1 gene was cloned and sequenced. Comparison of the proximal promoter region revealed a CpG island and two consensus sequences for the cAMP response element (CRE). The role of the PKA pathway in transactivation of the NPC 1 promoter was probed by transient transfection of a 2.2 kb fragment of the 5' flanking region fused to the luciferase reporter into mouse adrenal Y-1 and Leydig tumor cells and into nonsteroidogenic monkey kidney CV-1 cells. This promoter fragment displayed significant constitutive transcription, which was enhanced 3-5 fold in steroidogenic cells by treatment with 1 mM cAMP. The cAMP response was muted in CV-1 cells. We conclude that the NPC-1 gene is regulated by cAMP. It plays a role in normal cholesterol homeostasis and is essential for normal adrenal development and function. PMID- 12530643 TI - The obligatory action of protein tyrosine phosphatases in ACTH-stimulated steroidogenesis is exerted at the level of StAR protein. AB - A key regulatory step in the steroidogenic hormones signaling pathway is the synthesis of steroidogenic acute regulatory protein (StAR). This protein facilitates the delivery of cholesterol to the inner mitochondrial membrane, the rate-limiting step in steroidogenesis. ACTH and LH pathway also includes tyrosine dephosphorylation processes. Indeed, our previous studies have demonstrated that both hormones increase protein tyrosine phosphatase (PTP) activity by a PKA dependent mechanism and that the action of PTPs is required for the stimulation of steroid biosynthesis in adrenal and Leydig cells. In order to test the putative relationship between PTP activity and StAR protein induction in adrenocortical cells, in the present study we evaluated steroid production and StAR protein level in Y1 adrenocortical cells under PTP inhibition. Phenylarsine oxide (PAO), a powerful cell permeable PTP inhibitor, reduced ACTH-stimulated steroidogenesis in a concentration-dependent fashion. A concentration of 2.5 microM of this compound inhibited steroid synthesis in a 56% (ACTH = 318 +/- 30, ACTH + PAO = 145 +/- 18 ng progesterone/mL, P < 0.001) and also abrogated StAR protein induction. Phenylarsine oxide reduced the protein level after 60 min and this effect still remained at 120 min. A second PTP inhibitor, benzyl phosphonic acid, acting by a different mechanism, reproduced PAO effects on both steroidogenesis and StAR protein. Taken together, these results indicate that PTP activity participates in StAR protein induction and led us to attribute to the PKA-mediated PTP activation in steroidogenic systems a functional role, as mediator of StAR protein induction. PMID- 12530644 TI - Molecular modeling and structure-based thermodynamic analysis of the StAR protein. AB - Although much progress has been achieved in the study of the steroidogenic acute regulatory protein (StAR) dependent cholesterol transfer inside mitochondria, not one mechanism can account for all experimental data obtained to date. We have thus investigated the possibility that molecular modeling and structure-based thermodynamic calculations (STC) could enlighten these discrepancies. Starting from the crystallographic data of the human MLN64, a StAR homology model was generated and subjected to STC to verify the importance of StAR structural alterations for proper function. As expected, the model resembled the MLN64 crystal, although no binding site "tunnel" was obtained. Instead, a closed cavity was discovered, approximately the size and shape of cholesterol. This suggests that StAR does indeed require structural alterations to allow cholesterol binding, most evidently by the C-terminal alpha-helix above the U-shaped beta barrel. Through STC, it is shown that unfolding of this helix is probable and leads to a 2% subpopulation of partially unfolded StAR, supportive of both the intermembrane shuttle and the molten globule hypotheses. PMID- 12530645 TI - Cholesterol and steroid synthesizing smooth endoplasmic reticulum of adrenocortical cells contains high levels of translocation apparatus proteins. AB - Steroid-secreting cells possess abundant smooth endoplasmic reticulum whose membranes contain many enzymes involved in sterol and steroid synthesis. In this study we demonstrate that adrenal smooth microsomal subfractions enriched in these membranes also possess high levels of proteins belonging to the translocation apparatus, proteins previously assumed to be confined to morphologically identifiable rough endoplasmic reticulum (RER). We further demonstrate that these smooth microsomal subfractions are capable of effecting the functions of these protein complexes: co-translational translocation, signal peptide cleavage and N-glycosylation of newly synthesized polypeptides. We hypothesize that these elements participate in regulating the levels of ER targeted membrane proteins involved in cholesterol and steroid metabolism in a sterol-dependent and hormonally-regulated manner. PMID- 12530646 TI - The structure of microsomal cytochrome P450 2C5: a steroid and drug metabolizing enzyme. AB - The structure of microsomal P450 2C5 is the first structure of a membrane P450 to be determined by x-ray diffraction. This enzyme was originally identified as a progesterone 21-hydroxylase that is polymorphically expressed in rabbit liver. In contrast to the adrenal 21-hydroxylase, P450 2C5 metabolizes structurally diverse substrates that include a variety of steroids as well as therapeutic drugs. The flexible architecture of the enzyme and the residual solvation of the substrate provide a basis for understanding the catalytic diversity of 2C5 and related drug metabolizing P450s. In addition, the structure of P450 2C5 suggests how mammalian P450s have adapted for membrane binding and interaction with microsomal P450 reductase. PMID- 12530647 TI - Towards a unifying mechanism for CYP17 mutations that cause isolated 17,20-lyase deficiency. AB - Cytochrome P450c17 (CYP17) is a single hemoprotein that catalyzes both the 17alpha-hydroxylase and 17,20-lyase reactions in all species thus far examined. Severe defects in CYP17 cause classical 17-hydroxylase deficiency, but other defects result in partial or selective deficiency states. One such variant is the syndrome of isolated 17,20-lyase deficiency. Recent detailed studies of the biochemical properties of the mutant CYP17 enzymes from patients with isolated 17,20-lyase deficiency demonstrate that alterations in the interaction of CYP17 with its redox partner proteins P450-oxidoreductase and cytochrome b5 form the biochemical basis for these selective enzyme defects. Site-directed mutagenesis studies have confirmed that neutralization of any of several positive charges on the redox partner binding surface results in selective disruption of 17,20-lyase activity. In one case diagnosed as isolated 17,20-lyase deficiency, the identified mutation did not map to the redox partner binding surface; however, we have shown that this mutation cannot be the cause of isolated 17,20-lyase deficiency in this patient. These consistent results have prompted us to propose a paradigm in which neutralization of positive charges in the redox partner binding surface of CYP17 may be the predominant if not sole mechanism leading to isolated 17,20-lyase deficiency. PMID- 12530648 TI - 11Beta-hydroxysteroid dehydrogenase type 1 in differentiating omental human preadipocytes: from de-activation to generation of cortisol. AB - In humans, glucocorticoids are important regulators of adipose tissue distribution and function but circulating cortisol concentrations are normal in most patients with obesity. However, intracellular glucocorticoid levels can be modified by a microsomal enzyme 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) expressed mainly in the liver and adipose tissue. Locally generated cortisol within human adipose tissue can induce preadipocyte differentiation, but the relationship between 11beta-HSD1 expression and adipogenesis is unknown. Our present study has shown that in intact, undifferentiated omental (OM) but not subcutaneous (SC) preadipocytes, 11beta-HSD1 acts primarily as a dehydrogenase inactivating cortisol to cortisone. When preadipocytes become "committed" to adipocyte differentiation, oxo-reductase activity predominates generating cortisol. Since glucocorticoids are not only essential for OM preadipocyte differentiation but also inhibit cell proliferation, we postulate that 11beta HSD1 dehydrogenase activity in "uncommitted" OM preadipocytes may provide an autocrine mechanism to protect preadipocytes from differentiation, in turn facilitating their proliferation. Once early differentiation is initiated, a "switch" to 11beta-HSD1 oxo-reductase activity generates cortisol, thus promoting adipogenesis. The differences in set-point of 11beta-HSD1 activity between OM and SC human adipose tissue may be an important factor in the pathogenesis of visceral obesity. PMID- 12530649 TI - Synthesis, regulation, and function of neurosteroids. PMID- 12530650 TI - Difference of in vivo and in vitro antimineralocorticoid potency of progesterone. AB - Progesterone (P) given intramuscularly increases renal sodium excretion. We tested the in vitro capacity of P to bind to the human mineralocorticoid receptor (MR) with a reticulocyte lysate system and Ps transactivation potency in transfected CV-1 cells. Progesterone binds with higher affinity to the MR than aldosterone, but shows only low transactivation activity. This results in a very strong anti-mineralocorticoid (MC) potency of P in vitro. To test the in vivo anti-MC potency of P we infused aldosterone intravenously to hypo-MC Addison's patients, followed by increasing P infusions. During the study we measured normal aldosterone plasma concentrations and high P concentrations similar to the third trimester of pregnancy. Despite the 1000-fold higher plasma P concentrations, the in vivo anti-MC effect of P (increase of urinary sodium/potassium ratio) was rather small. We suggest that this may be due to effective MR protection mechanisms, such as conversion of P to inactive metabolites. PMID- 12530651 TI - Differences in substrate and inhibitor kinetics of human type 1 and type 2 3beta hydroxysteroid dehydrogenase are explained by the type 1 mutant, H156Y. AB - Two distinct genes encode the human type 1 (placenta, mammary gland) and type 2 (adrenal, gonad) isoforms of 3beta-hydroxysteroid dehydrogenase/isomerase (3beta HSD). We have produced the Y154F, H156Y, and K158Q mutant enzymes in the Y154-P H156-S-K158 motif of the human type 1 3beta-HSD/isomerase. The H156Y mutant was created to produce a chimera of the type 2 enzyme motif (Y154-P-Y156-S-K158) in the type 1 enzyme. The wild-type (WT) 1 and 2 plus the mutant enzymes were expressed and purified. The Km for dehydroepiandrosterone and Ki for epostane measured with both the H156Y mutant and WT 2 are 13-fold to 17-fold greater than those values obtained with the WT 1 3beta-HSD. The Y154F and K158Q mutants exhibit no 3beta-HSD but have significant isomerase activity. Thus, H156 in WT 1 vs. Y156 in WT 2 accounts for the substantially higher affinity of WT 1 3beta-HSD activity for these substrate and inhibitor steroids relative to the WT 2 enzyme. PMID- 12530652 TI - Baboon CYP11B1: the localization and catalytic activity in baboon adrenal tissue. AB - A third gene encoding baboon CYP11B1 was isolated and was shown to catalyze only the metabolism of deoxycorticosterone (DOC) to corticosterone. The investigation into the localization of CYP11B1 in the baboon adrenal tissue, using in situ hybridization, showed that mRNA transcripts were predominantly present in the zona reticularis (ZR) and zona fasciculata (ZF). Signal was also observed in the zona glomerulosa (ZG) and scattered within the medulla. Immunohistochemical studies, using rabbit anti-sheep CYP11B1 IgG, indicated that CYP11B1 was expressed only in the zona fasciculata, zona reticularis and in the medulla. CYP11B1 was not detected in the zona glomerulosa. Subsequent Western Blot investigations into the presence of CYP11B1 in baboon adrenal cortex and medullary homogenates indicated CYP11B1 as a single band in the cortex and as two distinct bands in the medulla. CYP11A was present only in the baboon adrenal cortex. The metabolism of deoxycorticosterone and corticosterone was subsequently investigated in the baboon adrenal cortex and medulla. In cortex homogenates, deoxycorticosterone was converted to corticosterone, and neither 18 hydroxycorticosterone nor aldosterone was detected. In medulla homogenates, however, corticosterone was metabolized to aldosterone, as confirmed by APcI-MS. PMID- 12530653 TI - Sheep adrenal cytochrome b5: active as a monomer or a tetramer in vivo? AB - Cytochrome b5 (cyt b5) is an ubiquitous hemoprotein also associated with microsomal cytochromes P450. It has been reported that cyt b5 influences cytochrome P450-dependent catalyses through electron transport as well as direct protein-protein interactions. To investigate the influence of cyt b5 on ovine adrenal steroidogenesis, we isolated and characterized cyt b5 from ovine liver. The molecular mass of the purified protein was 15,260 as determined by electrospray mass spectrometry. SDS-Polyacrylamide gel electrophoresis, even after stringent detergent and mercaptoethanol pretreatment, indicated multimeric forms of the protein, the most prominent being the tetramer (+/-60 kDa) with minor bands corresponding to the monomer (+/-16 kDa) and dimer (+/-30 kDa). Trypsin treatment of cyt b5 resulted in a truncated enzyme with a molecular mass of +/-10 kDa. The aggregation of cytochrome b5 was abolished by the tryptic removal of the membrane binding region. In Western blot analyses antibodies against the truncated protein recognised only this low molecular mass form and not the full length cyt b5, or any of the higher molecular complexes, showing the involvement of the membrane binding domain of the protein, not only in aggregation, but also in the quaternary structure which determines epitope presentation for antibody production. Immunoblot analyses of sheep adrenal microsomes with the anti-truncated cyt b5 antibody were also negative. Immunoblot analyses and immunocytochemistry of adrenal tissue with antibodies against the full length cyt b5 indicated that the tetrameric form of the protein was in all probability the dominant specie in vivo. PMID- 12530654 TI - Development of a transgenic green fluorescent protein lineage marker for steroidogenic factor 1. AB - Knockout (KO) mice lacking steroidogenic factor 1 (SF-1, officially designated Nr5a1) have a complex phenotype that includes adrenal and gonadal agenesis, impaired function of pituitary gonadotropes, and abnormalities of the ventromedial hypothalamic nucleus (VMH). To develop a lineage marker for cells that express SF-1, we used bacterial artificial chromosome (BAC) transgenesis. A BAC fragment containing 50 kb of the mouse Nr5a1 gene was placed upstream of the coding sequence for enhanced green fluorescent protein (eGFP) and used to generate SF-1/eGFP transgenic mice. These sequences directed eGFP expression to multiple cell lineages that express SF-1, including steroidogenic cells of the adrenal cortex, testes, and ovaries, VMH neurons, and reticuloendothelial cells of the spleen. Despite the essential role of SF-1 in gonadotropes, eGFP was not expressed in the anterior pituitary. These studies show that 50 kb of the mouse Nr5a1 gene can target transgenic expression to multiple cell lineages that normally express SF-1. The SF-1/eGFP transgene provides a valuable tool to expand our understanding of the actions of SF-1 in endocrine development and function. PMID- 12530655 TI - Differential regulation of SF-1-cofactor interactions. AB - The orphan nuclear receptor steroidogenic factor-1 (SF-1) plays pivotal roles in the development and function of steroidogenic organs. Here we describe the differential effect of protein kinase A (PKA) on coregulation of SF-1 dependent transcription by two p160 family members, p300/CBP co-integrator-associated protein (p/CIP) and transcription intermediary factor-2 (TIF2). Thus, whereas p/CIP-stimulated SF-1 dependent transcription is further potentiated by PKA, we show that activation of PKA leads to selective downregulation of TIF2 protein and a subsequent repression of TIF2 coactivator function. Using a yeast two-hybrid screen we also identified a novel zinc finger containing protein, which interacts with SF-1 via the AF-2 domain. PMID- 12530656 TI - SF-1 controls the expression of the scavenger gene akr1b7: in vitro and in vivo approaches. PMID- 12530657 TI - SF1 polymorphisms in the mouse and steroidogenic potential. AB - ACTH-resistance in four mutant derivatives of a mouse adrenocortical tumor cell line results from a defect that reduces the activity of steroidogenic factor-1 (SF1) thereby preventing expression of the ACTH receptor and other SF1-dependent genes. The SF1 genes from these mutants contain a sequence difference that changes an Ala to Ser at codon 172. Steroidogenic factor-1(S172) represents a polymorphism rather than a spontaneous mutation since the two forms of SF1, SF1(A172), and SF1(S172), can be traced to the hybrid mouse strain (C57L/J x A/HeJ) from which the original adrenal tumor was derived. The SF1(S172) allele is amplified in three of the four mutant clones together with the neighboring genes germ cell nuclear factor and LIM homeobox2. The two forms of SF1 had only modest differences in transcriptional activity in reporter gene assays, suggesting that the SF1 polymorphism per se is not directly responsible for the loss of mc2r expression. Rather, ACTH resistance in this family of adrenocortical tumor cell mutants may be due to a closely linked gene on the SF1(S172) allele. Mouse strains with reportedly high steroidogenic capacity (C57Bl/6J, C57Bl/10J) also have the SF1(A172) allele while mouse strains with low steroidogenic capacity (C3H/HeJ, DBA/2J) have the SF1(S172) allele. These latter observations suggest that the two SF1 alleles also may be markers of steroidogenic potential among mouse strains. PMID- 12530658 TI - Role of three SF-1 binding sites in the expression of the mvdp/akr1-b7 isocaproaldehyde reductase in Y1 cells. AB - Mvdp/akr1-b7 encodes an aldose-reductase-like enzyme expressed in the zona fasciculata of the adrenal cortex, the function of which is essential for the detoxification of the cholesterol side chain cleavage product, isocaproaldehyde. The -510/+41 akr1-b7 promoter fragment is able to reproduce the endogenous gene zona fasciculata restricted, ACTH-controlled expression, in transgenic mice adrenals. Here, we report that three response elements contained within this promoter (positions -102, -458, -503) are able to bind SF-1, the essential regulator of steroidogenesis, although the low affinity site at -503 retains some other specific proteins present in Y1 nuclear extracts. Mutation of the -102 site results in a lowering of the activity of the -510/+41 promoter in Y1 cells, whereas mutation of the -458 site induces a reduction both in the global activity and forskolin sensitivity of the promoter. Interestingly, differential mutations of the -503 site nucleotides either induce an increase or a decrease in the basal and forskolin-induced activity. PMID- 12530659 TI - Analysis of Ad4BP/SF-1 gene regulatory region. PMID- 12530660 TI - LXXLL motifs in Dax-1 have target specificity for the orphan nuclear receptors Ad4BP/SF-1 and LRH-1. PMID- 12530661 TI - Knock-ins and conditional knockouts: in vivo analysis of glucocorticoid receptor regulation and function. AB - To determine the cellular targets for glucocorticoid (GC) action, we have generated mice in which a green fluorescent protein-glucocorticoid receptor (GFP GR) fusion gene is knocked into the endogenous GR locus. We found that GFP-GR function is indistinguishable from endogenous GR on both a cellular and systemic level. Furthermore, the green fluorescence intensity of the GFP-GR protein is proportional to its expression, allowing quantitation of GR expression in single living cells. We initiated our analysis of GR regulation in the thymus. Using multicolor flow cytometry, we found that GR expression is uniform among embryonic thymocyte subpopulations, but gradually "matures" over a three-week period after birth. In the adult, analysis of GFP-GR expression on RAG2-/- and HY T cell receptor (TCR) transgenic genetic backgrounds, showed that GR is induced to high levels in immature CD25+ CD4- CD8- thymocytes and down-regulated by activation of the pre-TCR during positive but not negative selection. Additionally, relative GR expression is dissociated from GC-induced apoptosis in vivo. These results implicate pre-TCR signaling as a mechanism for GR down-regulation and separate receptor abundance from susceptibility to apoptosis across thymocyte populations. PMID- 12530662 TI - Transcriptional complexes at the CYP17 CRS. AB - Steroid hormone biosynthesis in the adrenal cortex is controlled by the peptide hormone adrenocorticotropin (ACTH), which acts to increase intracellular cAMP, resulting in the activation of cAMP-dependent protein kinase (PKA) and subsequent increase in steroidogenic gene transcription. We have identified three proteins interacting with the human CYP17 cAMP responsive sequence (CRS): steroidogenic factor 1 (SF-1), p54nrb, and polypyrimidine tract-binding protein-associated splicing factor (PSF). Nuclear extracts isolated from cAMP stimulated of H295R cells showed cAMP-inducible binding to the human CYP17 (hCYP17) CRS. This cAMP inducible binding was dependent on a dual-specificity phosphatase (DSP). DSP activity was subsequently shown to be is essential for conveying ACTH/cAMP stimulated transcription of several steroidogenic genes in the human adrenal cortex. We report here that the transactivation potential of SF-1 is also dependent on phosphatase activity; suggesting that SF-1 is dephosphorylated in response to ACTH/cAMP stimulation. Finally, we demonstrate a role for mitogen activated protein kinase phosphatase 1 (MKP-1), a nuclear DSP, in conveying SF-1 dependent transcription of an hCYP17 promoter-reporter construct in the H295R human adrenocortical cell line. We conclude that a DSP, possibly MKP-1, is essential for enhancing hCYP17 transcription in the adrenal cortex by desphosphorylating of SF-1, thereby increasing the binding affinity of SF-1, p54nrb, and PSF for the hCYP17 promoter. PMID- 12530664 TI - Steroid deficiency syndromes in mice with targeted disruption of Cyp11a1. PMID- 12530663 TI - Function of the transcriptional regulating protein of 132 kDa (TReP-132) on human P450scc gene expression. AB - Cytochrome P450scc catalyzes the important first step in the steroid synthesis pathway; however, it is clear that additional factors regulating the temporal and spacial specific expression of the CYP11A1 gene remain to be identified. To isolate novel transcription factors that regulate this gene, a cis-acting element of the 5'-flanking region from nucleotides -155 to -131 (-155/-131) was used to screen a human placental lambda gt11 cDNA expression library, and an interacting clone was isolated. The open reading frame of the cDNA encodes several domains that are characteristic of transcription factors including an acidic region, a region rich in prolines and three zinc-finger motifs. Expression of the cDNA by in vitro transcription/translation and by transient transfection in HeLa cells yielded a protein of 132 kDa, which concurs with the predicted size. Transfection of the cDNA in placental JEG-3 and adrenal NCI-H295 cells, stimulate expression of a reporter construct controlled by the P450scc gene 5'-flanking region from nucleotides -1676 to +49. This transcriptional regulating protein of 132kDa (TReP 132) when expressed in HeLa cells was demonstrated to interact with the -155/-131 region in bandshift analysis, and tandem copies of this region was shown to confer activation of the heterologous HSV thymidine kinase minimal promoter. Coexpression of CBP/p300 with TReP-132 further increased promoter activity, and the proteins were demonstrated to interact physically. RNA analysis demonstrated the highest levels of expression in the adrenal cortex and testis; and transcript expression is also found in the steroidogenic JEG-3, NCI-H295, and MCF-7 cell lines, but not in non-steroidogenic HepG2 and HK293 cells. Subsequently it has been shown that TReP-132 interacts with steroidogenic factor-1 (SF-1) through specific domains; and along with the interaction with CBP/p300 these factors are postulated to form a complex to regulate expression of the P450scc gene. PMID- 12530665 TI - Partial characterisation of the 5' flanking region of the human GIP receptor (GIPR) gene. PMID- 12530666 TI - CIP-1 is a novel corepressor for nuclear receptor COUP-TF: a potential negative regulator in adrenal steroidogenesis. PMID- 12530667 TI - A ring-finger protein CIP-2 is a novel regulator of COUP-TF action in the adrenal cortex. PMID- 12530668 TI - Adrenal growth is controlled by expression of specific pro-opiomelanocortin serine protease in the outer adrenal cortex. AB - As previous work had shown that extreme N-terminal fragments of the ACTH precursor pro-opiomelanocortin (POMC) not containing gamma-melanotropin (gamma MSH) were active adrenal mitogens but an antiserum raised against gamma-MSH paradoxically also inhibited adrenal growth we proposed that the adrenal mitogen is processed from pro-gamma-MSH by a neurally controlled protease at the growing adrenal. To this end we have characterised a novel serine protease (named adrenal secretory protease (AsP) as Psort predicted a leader motif) which is expressed at the glomerulosa/fasciculata boundary where mitosis takes place. The expression of AsP was also found to be essential for mitosis of the adrenal cortical tumor Y1 cell-line in POMC containing media and 3D homology modeling revealed the presence of a catalytic pocket flanked by the classical His/Asp/Ser motifs. An usual feature of the model was a cluster of arginine residues on the underside of the protease suggesting that this basically charged face would tend to retain it on the cell surface on secretion-immunocytochemistry using an antiserum raised against a synthetic peptide spanning residues 1-25 of AsP showed that this was the case for Y1 cells. Specificity of AsP (affinity purified from Y1 media) was demonstrated by its inability to cleave model substrates for either trypsin or pro-hormone converting enzymes but was able to cleave an internally quenched POMC (44-55) model peptide. Interestingly mass spectral analysis of products of the latter predicts that the protease cleaves between the bond between Val52 and Met53 suggesting the natural adrenal mitogen is POMC (1-52). PMID- 12530669 TI - SF-1, DAX-1, and acd: molecular determinants of adrenocortical growth and steroidogenesis. AB - The formation of the adrenal cortex in humans is notable for the presence of two discrete zones, the fetal zone (FZ) which regresses soon after birth and the definitive zone (DZ) which gives rise to the classic steroidogenic zones of the adult cortex. Mice possess an analogous structure to the FZ referred to as the X zone (XZ) which regresses at puberty in the male and during the first pregnancy in the female. Similar to the human FZ in X-linked Congenital Adrenal Hypoplasia caused by loss of function mutations in DAX-1 (Dosage-sensitive sex reversal Adrenal hypoplasia congenita critical region on the X chromosome), the mouse XZ does not regress when DAX-1 is mutated. Only in humans with DAX-1 mutations, however, is the DZ small and hypofunctional. Patients and mice with SF-1 mutations have complete adrenal aplasia with absence of both the DZ and FZ/XZ. Lastly, the phenotype of the Autosomal Recessive Adrenocortical Dysplasia (acd) mouse is strikingly similar to human Miniature Adult Congenital Adrenal Hypoplasia, lacking an XZ/FZ and possessing a dysfunctional DZ. Current work has addressed the regulation of SF-1 and DAX-1 dependent adrenocortical growth and steroidogenesis in vivo utilizing mouse models of simple and combined SF-1 and DAX-1 deficiency. In addition, the model of compensatory adrenal growth in SF-1 haplo-insufficient mice has been applied to evaluate the potential role of SF-1 in adrenocortical proliferation. Additional efforts aim to positionally clone the acd gene, predicated on the hypothesis that it is a critical component of the adrenal developmental cascade. PMID- 12530670 TI - Molecular genetics of adrenal hypoplasia congenita. PMID- 12530671 TI - The adrenal genetic puzzle: how do the fetal and adult pieces differ? AB - The basis for the steroidogenic differences between the human fetal adrenal (HFA) and adult adrenal is not well defined. However, gene expression clearly plays a critical role in defining their distinct steroidogenic and structural phenotypes. We used DNA microarrays to compare expression levels of several thousand transcripts between the HFA and adult adrenal gland. Gene profiling was done using seven independent microarrays that contained between 7075 and 9182 cDNA elements. Twenty-five transcripts were found to have a greater than 5-fold difference in expression between HFA and adult adrenals. The largest differences were observed for transcripts that encode insulin-like growth factor-II (IGF-II) (25-fold higher in HFA) and 3beta hydroxysteroid dehydrogenase (3betaHSD) (21 fold higher in adult). The vast majority of the 25 transcripts have not been studied with regard to adrenal function. We also determined the transcripts that had the highest signal intensities, which is an approximate measure of expression level, for both the fetal and adult adrenal RNA samples. The enzyme 24 dehydrocholesterol reductase, which is involved in cholesterol biosynthesis, exhibited the highest signal intensity for fetal adrenal RNA. For adult adrenal mRNA, the expression of 11beta-hydroxylase transcripts was found to have the highest signal intensity ranking. Overall, 10 of the top 20 highest signal intensities were similar for adult and fetal adrenal transcripts. The gene profile data for fetal vs. adult adrenal glands should provide valuable information that could help define mechanisms involved in adrenal growth and development. PMID- 12530672 TI - Cytochrome b5 (b5) in the developing rhesus adrenal. PMID- 12530673 TI - Effect of N-proopiomelanocortin (1-77) and (1-49) infusions on adrenal expression of cyclin D1 in the fetal sheep. AB - In the sheep, there is a rapid increase in fetal adrenal growth and steroidogenesis during the last 10-15 days gestation. Recently, we have shown that infusion of POMC 1-77 increases fetal adrenal growth but does not significantly alter fetal plasma cortisol concentrations. Phosphorylation and inactivation of the pRB protein, which is required for progression into the DNA synthetic phase of the cell-cycle is conducted by a holoenzyme, for which cyclin D1 gene encodes the rate-limiting regulatory subunit. To further elucidate the mechanisms by which POMC 1-77 regulates adrenal growth, we therefore examined adrenal expression of the rate-limiting cell cycle protein, cyclin D1, from fetuses infused for 48 hr with POMC 1-77 (n = 6), POMC 1-49 or Saline (n = 6). There was no significant difference in the adrenal expression of cyclin D1 mRNA levels between POMC 1-77, 1-49 and saline infused fetuses. There was no significant correlation between cyclin D1 (4.0 Kb) and adrenal weight. In summary, these data do not demonstrate that the rate-limiting cell cycle protein, cyclin D1, is activated to stimulate adrenal growth following infusion of POMC 1 77 in the fetal sheep in late gestation. PMID- 12530674 TI - Expression of the melanocortin receptors MC2-R (ACTH-receptor) and MC5-R during embryonic development of ovine adrenals. AB - Two melanocortin receptors MC2-R (ACTH-receptor) and MC5-R are expressed in the adult lamb adrenal cortex. In this work, we have studied the time-course of expression of these two receptors during ovine fetal development. MC2-R expression progressively increases from day 60 to day 140 of gestation (x3), then more rapidly before parturition and remains constant in the newborn. In contrast, the pattern of MC5-R expression is totally different. A strong increase is observed between days 60 and 120 (x7) then followed by a decrease until parturition and after birth. This peak of MC5-R expression precedes that of MC2 R, suggesting that MC5-R might be involved in alpha-MSH- and/or ACTH-stimulated corticosteroid synthesis during early embryonic life. PMID- 12530675 TI - Fibronectin, laminin, and collagen IV interact with ACTH and angiotensin II to dictate specific cell behavior and secretion in human fetal adrenal cells in culture. AB - Whereas collagen IV is expressed throughout the fetal adrenal gland during the second trimester of human development, fibronectin, and laminin demonstrate a rather mirror-image distribution, with higher expression of fibronectin in the central portion and laminin at the periphery of the gland. In the present study, extracellular matrices were able to modulate the profile of steroid secretion in primary cultures: collagen IV favored cortisol secretion following adrenocorticotropin (ACTH) or angiotensin II (Ang II) stimulation while specific stimulation of the AT2 receptor of Ang II elicited dehydroepiandrostenedione (DHEA) production. These effects were correlated by changes in mRNA levels of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and cytochrome P450C17. In contrast, fibronectin and laminin decreased cell responsiveness to ACTH in terms of cortisol secretion, but enhanced ACTH-stimulated androgen secretion. Finally, extracellular matrices were able to orchestrate cell behavior: collagen IV and laminin enhanced cell proliferation whereas fibronectin incited cell death. These results indicate that the nature of extracellular matrix coordinates specific steroidogenic pathways and cell turnover in the developing human fetal adrenal gland. PMID- 12530676 TI - Steroidogenic enzyme expression within the adrenal cortex during early human gestation. AB - Aberrant adrenocortical function during the first trimester of human fetal development underlies the severe virilization of congenital adrenal hyperplasia due to cytochrome P450 21-hydroxylase (CYP21) deficiency. Although valuable information of human adrenocortical development after 12 weeks gestation is available, less is known earlier in pregnancy. In our studies, the adrenal cortex was first detected in human embryos by hematoxylin and eosin staining at 33 days post-conception (dpc) with distinction between the definitive and fetal zones possible at 52 dpc. Vascular development was apparent within the adrenal gland at 41 dpc. CYP11A and CYP17 were expressed centrally within the fetal zone at 50 dpc and all later time points during the first trimester. Weaker CYP11A immunoreactivity also was visible in the outer region of the adrenal cortex consistent with definitive zone expression. In this location, immunoreactivity was observed for 3beta-hydroxysteroid dehydrogenase and the proliferation marker, Ki67. These data raise the possibility of de novo cortisol biosynthesis during the first trimester of human development and are relevant to the pathophysiology of 46,XX virilization in CYP21 deficiency. PMID- 12530677 TI - Transcription factors GATA-4 and GATA-6 during mouse and human adrenocortical development. AB - Our earlier work implicates transcription factors GATA-4 and GATA-6 in the murine adrenal. We have now studied their expression during mouse and human adrenal development in detail. GATA-4 and GATA-6 mRNAs are readily detectable from embryonic day 15 in mouse and gestational week 19 in human adrenal cortex. In postnatal adrenal, GATA-4 expression is down-regulated, whereas GATA-6 mRNA and protein continue to be abundantly present. In a human adrenocortical cell line NCI-H295R, GATA-6 mRNA is up-regulated by cAMP. This cell line does not express GATA-4. Our findings suggest that GATA-6 expression is hormonally controlled, and required throughout adrenal development from fetal to adult age. GATA-4, on the other hand, may serve a role in fetal adrenal gene regulation. PMID- 12530678 TI - Involvement of T-type calcium channels in the mechanism of action of 5-HT in rat glomerulosa cells: a novel signaling pathway for the 5-HT7 receptor. AB - We have previously demonstrated that, in rat, the stimulatory effect of 5-HT on aldosterone secretion is mediated through a 5-HT7 receptor subtype. The aim of the present study was to characterize the transduction mechanisms associated with activation of native 5-HT7 receptors. 5-HT induced a dose-dependent increase in cAMP production in rat glomerulosa cells. Pretreatment of cells with the adenylyl cyclase (AC) inhibitor SQ 22536 or the protein kinase A (PKA) inhibitor H-89 markedly attenuated the effect of 5-HT on aldosterone secretion. Administration of 5-HT in the vicinity of glomerulosa cells induced a robust increase in cytosolic calcium concentration ([Ca2+]i) and this effect was abrogated by the T type calcium channel blocker mibefradil. Patch-clamp studies confirmed that 5-HT activated a T-type calcium current. H-89 attenuated both the [Ca2+]i response and the activation of T-type calcium current induced by 5-HT. Reduction of extracellular calcium concentration in the medium or administration of mibefradil caused a marked reduction of the maximum effect (Emax) of 5-HT on aldosterone secretion. These data demonstrate that activation of native 5-HT7 receptors stimulates cAMP formation, which in turn provokes calcium influx through T-type calcium channels. Both the activation of the AC/PKA pathway and the calcium influx are involved in 5-HT-induced aldosterone secretion. PMID- 12530679 TI - Mosaic patch patterns in chimeric and transgenic mice suggest that directional growth in the adrenal cortex begins in the perinatal period. AB - Staining for beta-galactosidase (beta-gal) reporter activity in adrenal glands from adult, fetal and neonatal 21-OH/LacZ transgenic mice revealed mosaic patch patterns that were qualitatively similar to those seen in LacZ <--> wild-type mouse chimeras, at similar developmental stages. This suggests that, as in chimeras, the transgenic patch pattern may reflect cell lineage relationships. Consequently, 21-OH/LacZ transgenic mice could be useful as a simpler alternative to chimeras for the analysis of clonal growth and cell mixing during adrenocortical organogenesis. Embryonic day 16.5 (E16.5) adrenal cortices of 21 OH/LacZ transgenic mice displayed a punctate patch pattern, but by E18.5 "stripes" appeared to be emerging and were clearly visible by the day of birth (P0) and three days later (P3), consistent with the adult mosaic striped pattern. This suggests that adrenocortical organogenesis involves an initial period of randomly oriented clonal growth, followed by directional growth which begins in the perinatal period. PMID- 12530680 TI - Redistribution of connexin 43 by cAMP: a mechanism for growth control in adrenal cells. AB - In order to investigate the regulation of connexin distribution in rapidly growing cells, we evaluated the effects of dibutyryl cyclic adenosine monophosphate (DbcAMP) treatment on the cellular location of endogenous connexin 43 (Cx43) in an adrenal tumor cell population. Polyclonal antibodies to Cx43 and a Cx43 green fluorescent fusion protein (Cx43-GFP) were used to localize Cx43 gap junction protein in plaques at sites of cell-cell contact and in spherical packets within the cytoplasm. Treatment with DbcAMP significantly decreased the number and size of packets within the cytoplasm, and significantly increased surface gap junction plaque size. These data support the theory that cAMP can influence the availability of gap junction protein at the points of cell contact where it is required for cell-cell communication, and thus has the potential to be an important regulator of adrenal tumor cell proliferation. PMID- 12530681 TI - Hox gene expression in adult tissues with particular reference to the adrenal gland. AB - In comparison to the embryo, very little work has been carried out on the expression and role of Hox genes in the adult animal. An expression profile of all 39 vertebrate Hox genes on a select panel of adult human tissues reveals that in fact these genes are widely expressed throughout the adult human and a colinear pattern of expression is displayed similar to that of the developing embryo. Of particular interest is the abundance of Hox genes that are expressed within the adult adrenal gland. Adrenal cortical cells are continuously renewed to sustain production of zonal steroids. Cell proliferation occurs at the periphery of the cortex and cells are then displaced centripetally, phenotypically switching as they migrate through the gland before undergoing apoptosis at the zona reticularis/medullary boundary. It is still unclear which mechanisms cause the cells to differentiate as they cross the zonal boundaries and we hypothesise that Hox genes may be involved in the phenotypic switching of the adrenocortical cells. In situ hybridisation experiments were carried out on adult rat adrenal gland sections and Hox gene expression was localized within the zonal borders, coinciding with the localization of cells that undergo phenotypic differentiation, and thus supporting our hypothesis that Hox genes may be involved in the phenotypic switching of the adrenocortical cells. As in the developing embryo, the genes display colinear expression with the 3' Hox genes being expressed within the outer gland and the 5' genes within the inner zones. PMID- 12530682 TI - Marmosets express a fetal zone at birth but no ZR in adulthood. PMID- 12530683 TI - Neuropeptides and adrenocortical proliferation in vitro. AB - Vasoactive intestinal peptide (VIP) and Neuropeptide Y (NPY) are localised in the capsule and zona glomerulosa region of the adrenal cortex, where they play an important role in regulating steroidogenesis and adrenal blood flow. This study investigates the effect of these neuropeptides on adrenocortical cellular proliferation and steroidogenesis in vitro. Capsular/glomerulosa and innerzone/medulla preparations were either stimulated acutely with NPY or VIP (both 10(-6) M) for up to 2 hours or for 24 hours, four and eight days in vitro in eagles MEM (3.4 mM K+). DNA synthesis was determined using immunocytochemistry through the incorporation of the thymidine analogue 5-bromo-2'-deoxyridine (BrdU, 20 mg/mL). Phosphorylation of mitogen activated protein kinase ERK1/2 was assessed by western blotting. Both VIP (10(-6) M) and NPY (10(-6) M) treatment caused an increase in DNA synthesis after four days in culture. Acute NPY treatment caused an increase in ERK1 and 2 phosphorylation (p < 0.01) in the capsular/zona glomerulosa. Vasoactive intestinal peptide treatment caused a significant increase in ERK 1/2 phosphorylation (p < 0.05) only in innerzones/medulla preparations. Both responses were maximal between 10 and 30 min of incubation and decrease thereafter. These data provide further evidence for the role of the mitogen activated protein kinases ERK1 and 2 in the proliferative events in the adrenal gland and demonstrate stimulation of cell division by the adrenal neuropeptides VIP and NPY in vitro. PMID- 12530684 TI - Proliferation of capsular stem cells induced by ACTH in the rat adrenal cortex. AB - Despite great efforts devoted to clarifying the localization of proliferative activity in the adrenal cortex, the agents that stimulate proliferation remain controversial, and the nature of the stem cells from which cortical cells differentiate is incompletely understood. We studied proliferative activity in the rat adrenal cortex using an immunohistochemical method to detect the presence of the Proliferating Cell Nuclear Antigen (PCNA) (an intranuclear enzyme whose synthesis reaches the maximum intensity during the S-phase of the cell cycle). Groups of six rats were subjected to daily intraperitoneal injection of either corticotropin (ACTH1-24--0.2 mg/kg), dexamethasone (Dexa--4 mg/kg) or 0.9% saline for three consecutive days and killed 24 h after the last injection. Adrenal weight was significantly increased by ACTH treatment and reduced by Dexa. Concentrations of endogenous ACTH in plasma were lower in the Dexa group than in controls, and curiously, this was true in the ACTH1-24 treated group as well, probably in consequence of the increased corticosterone levels providing negative feedback at the hypothalamic-pituitary level. Corticosterone levels, as expected, were increased by the ACTH stimulus and reduced by the use of Dexa. Proliferating cell nuclear antigen immunostaining was close to zero in Dexa treated animals and low in controls. In ACTH treated rats, a significantly increased number of cells were positively stained. Positive cells were identified in both in zona glomerulosa (ZG) and zona intermedia (ZI) but many were located in the capsule. Zona fasciculata (ZF) and zona reticularis (ZR) were devoid of staining in all of these cases. We conclude that pharmacological doses of ACTH induce proliferation of capsular fibroblasts. Following descriptions by early 20th century researchers it is possible that these cells may also be stem cells and differentiate into adrenal cortex cells. PMID- 12530685 TI - From maternal care to gene expression: DNA methylation and the maternal programming of stress responses. PMID- 12530686 TI - 11Beta-hydroxysteroid dehydrogenases: a novel control of glucocorticoid action in the brain. AB - Clearly the actions of 11beta-HSDs in the CNS are only beginning to be unraveled, though some principles are emerging. 11Beta-hydroxysteroid dehydrogenase type 2 inactivates glucocorticoids but is found only in a few very restricted sites in the adult brain. Its role in the fetal CNS is likely to be more extensive, but is largely unexplored. 11Beta-hydroxysteroid dehydrogenase type 1 is a reductase in CNS cells and amplifies glucocorticoid action. The striking protection against cognitive decline seen in elderly 11beta-HSD1 knockout mice suggest that manipulation of this isozyme may be a useful therapeutic avenue to explore. PMID- 12530687 TI - Glucocorticoids, hypothalamo-pituitary-adrenal (HPA) development, and life after birth. AB - Approximately 10% of women in North America are treated with synthetic glucocorticoid (sGC) between 24 and 32 weeks of pregnancy (term approximately 40 weeks), to promote lung maturation in fetuses at risk of preterm delivery. Such therapy is highly effective in reducing the frequency of respiratory complications, and as a result, repeated course treatment has become widespread. Nothing is known about the impact of repeated sGC treatment on neuroendocrine development in the human, or if specific time windows of increased sensitivity exist. Glucocorticoids are essential for many aspects of normal brain development. However, there is growing evidence from a number of species, that exposure of the fetal brain to excess glucocorticoid can have life-long effects on behaviour and neuroendocrine function. We have shown that exposure of fetuses to sGC in late gestation permanently alters HPA function in pre-pubertal, post pubertal, and aging offspring, in a sex-dependent manner. These effects are linked to changes in central glucocorticoid feedback. Prenatal glucocorticoid exposure also leads to modification of HPA-associated behaviours and organ morphology, as well as altered regulation of other neuroendocrine systems. Permanent changes in HPA function will have a long-term impact on health, since elevated cumulative exposure to endogenous glucocorticoid has been linked to the premature onset of pathologies associated with aging. PMID- 12530688 TI - The adrenal hormone metabolism in the immune/inflammatory reaction. AB - The interactions between the immune system and HPA axis may be characterized by a circuit which includes; (i) activation of the HPA axis and initiation of the stress response which, in term, has immunomodulating properties; (ii) a feedback mechanism derived from the immune system which regulates the HPA axis. Over the past few years, it has become evident that the adrenal gland, itself, as the main effector organ of the HPA axis, is a major site for both the synthesis and action of numerous cytokines. In addition to the cytokine mediated activation of adrenal regulation there are cytokine independent cell-cell mediated immune-adrenal interactions. The nature of this immune-endocrine crosstalk is implicated in adrenal dysfunction and disease. During inflammatory and autoimmune disorders including sepsis, inflammatory bowel disease and rheumatoid arthritis, the immune adrenal crosstalk becomes more critical in maintaining an adequate adrenal stress response. PMID- 12530689 TI - New insights into the molecular basis of the triple A syndrome. AB - The triple A syndrome (MIM*231550) is a rare autosomal recessive disorder characterized by adrenocorticotropic hormone (ACTH) resistant adrenal failure, achalasia, alacrima and a variety of neurological and dermatological features. Adrenal insufficiency usually presents in the first decade of life, however in some patients it may occur later in life or may even lack completely. Recently, we and others identified a novel gene on chromosome 12q13, designated AAAS (Achalasia-Addisonianism-Alacrima-Syndrome gene) which is mutated in patients with triple A syndrome. We investigated n=84 families including 111 patients with clinically suggested triple A syndrome and identified homozygous or compound heterozygous AAAS mutations in 78 families. Genotype/phenotype analyses revealed a highly variable occurrence, age of onset and severity of all clinical symptoms between patients with the same AAAS mutation. The obvious lack of a genotype/phenotype relationship is suggestive of modifying genes/factors which need to be determined. The AAAS protein function is unknown. With four WD repeats it belongs to the family of WD repeat-containing proteins which may exhibit a high degree of functional diversity. The subcellular localization of the protein and the determination of its putative binding partners will shed light on the role of the AAAS protein for the development and function of the adrenal gland and other neuroendocrine structures. PMID- 12530690 TI - Functional characterization of the adrenoleukodystrophy protein (ALDP) and disease pathogenesis. AB - X-linked adrenoleukodystrophy (X-ALD) is the most common peroxisomal disorder characterized by abnormal accumulation of saturated very long chain fatty acids in tissues and body fluids with predominance in brain white matter and adrenal cortex. The clinical phenotype is highly variable ranging from the severe childhood cerebral form to asymptomatic persons. The responsible ALD gene encodes the adrenoleukodystrophy protein (ALDP), a peroxisomal integral membrane protein that is a member of the ATP-binding cassette (ABC) transporter protein family. The patient gene mutations are heterogeneously distributed over the functional domains of ALDP. The extreme variability in clinical phenotype, even within one affected family, indicates that besides the ALD gene mutations other factors strongly influence the clinical phenotype. To understand the cell biology and function of mammalian peroxisomal ABC transporters and to determine their role in the pathogenesis of X-ALD we developed a system for expressing functional ABC protein domains in fusion with the maltose binding protein. Wild type and mutant fusion proteins of the nucleotide-binding fold were overexpressed, purified, and characterized by photoaffinity labeling with 8-azido ATP or 8-azido GTP and a coupled ATP regenerating enzyme assay for ATPase activity. Our studies provide evidence that peroxisomal ABC transporters utilize ATP to become a functional transporter and that ALD gene mutations alter peroxisomal transport function. The established disease model will be used further to study the influence of possible disease modifier proteins on ALDP function. PMID- 12530691 TI - Protein kinase A and tumorigenicity: the example of micronodular adrenocortical hyperplasia and Carney complex. PMID- 12530692 TI - Role of local 11beta-hydroxysteroid dehydrogenase type 2 (HSD11B2) expression in determining the phenotype of adrenal adenomas. PMID- 12530693 TI - Expression of ACTH receptor pathway genes in GIP-dependent Cushing's syndrome (CS). PMID- 12530694 TI - Analysis of the putative promoter region of the GIP receptor gene (GIPR) in GIP dependent Cushing's syndrome (CS). PMID- 12530695 TI - Endocrine gland vascular endothelial growth factor (EG-VEGF) and the hypothesis of tissue-specific regulation of angiogenesis. PMID- 12530697 TI - Adrenocortical cell proliferation in a cell transplantation model: the role of SV40 T antigen. AB - Bovine adrenocortical cells immortalized by human telomerase reverse transcriptase (hTERT) are capable of forming functional vascularized tissue structures when transplanted in immunodeficient mice. These tissues maintain the life of adrenalectomized animals, show normal cell proliferation rates, and maintain a constant tissue size. These experiments were performed by co transfection of an hTERT-encoding plasmid with an SV40 T antigen-encoding plasmid, but in tissues formed from clones derived in this way SV40 T Ag was not expressed. However, when tissues were formed from nonclonal heterogeneous populations of transfected cells, nodules of SV40 TAg-expressing cells arose that showed a high proliferation rate. These structures resembled nodules formed from transplanted bovine adrenocortical cells in which SV40 TAg was introduced by retroviral transduction rather than transfection. The reasons for these phenomena are discussed; in a nonclonal mixed population of cells, some may have much higher levels of SV40 TAg, which perturbs the normal histology and behavior of tissues formed from hTERT-immortalized adrenocortical cells. PMID- 12530696 TI - cAMP pathway alterations from the cell surface to the nucleus in adrenocortical tumors. AB - The cyclic AMP (cAMP) pathway plays a major role in the development of endocrine tissues and various molecular defects of key components of this pathway (G protein, receptors, PKA, ...) have been observed in endocrine tumors. Hypersecretion of adrenocorticotropin hormone (ACTH), the key activator of the cAMP pathway in adrenal cortex, is associated with adrenocortical hyperplasia and cortisol oversecretion (Cushing's syndrome). The best example of "illegitimate" membrane receptors expression reported is the abnormal expression of the adenylyl cyclase activating gastric inhibitory peptide receptor (GIP-R) in ACTH independent Cushing's syndrome (ACS). We have observed that ectopic expression of the GIP-R is frequent in ACTH-Independent Macronodular Adrenal Hyperplasia (AIMAH), rare in benign adrenal adenoma (AA), but seems absent in Adrenal Cancer (AC). In vivo systematic screening of AIMAH shows at least one abnormal response of cortisol (suggesting "illegitimate" membrane receptor expression) in almost all patients. Somatic and germ line inactivating mutations of PRKAR1 (regulatory subunit R1A of PKA) can be observed in patient with isolated primary pigmented nodular adrenocortical disease (PPNAD) and AA responsible for ACS. At the nuclear level, the cAMP pathway regulates transcription mainly by PKA-dependent phosphorylation of the cyclic AMP response element binding (CREB) family of transcription factors (CREB, CREM, and ATF-1). Cyclic AMP response element binding protein (CREB) is expressed in normal adrenal cortex. Alterations of CRE binding proteins with loss of CREB expression and compensatory overexpression of CREMtau is observed in the human adrenocortical cancer cell line H295R. Similar alterations are found at the protein level in human malignant adrenocortical tumors. In conclusion, various alterations leading to activation or inactivation of key components of the cAMP signaling pathway can be observed in adrenocortical tumorigenesis. PMID- 12530698 TI - Transcription profiling of benign and malignant adrenal tumors by cDNA macro array analysis. PMID- 12530699 TI - Effects of serotonin and vasopressin on cortisol production from an adrenocortical tumor causing subclinical Cushing's syndrome. AB - In dexamethasone-suppressed healthy volunteers, the serotonin4 (5-HT4) receptor agonist cisapride and lysine vasopressin [LVP, an analog of arginine vasopressin (AVP)] have no influence on plasma cortisol levels (PCL). In contrast, cisapride and AVP have been shown to stimulate cortisol secretion in patients with adrenal tumor or bilateral adrenal hyperplasia and Cushing's syndrome. In this report, we describe a case of adrenocortical adenoma causing subclinical Cushing's syndrome. Cisapride and terlipressin, a precursor of LVP, both induced an increase in PCL reaching +88% and +100%, respectively, without any significant variation of plasma ACTH levels. In vitro experiments were conducted to investigate the effects of 5-HT and AVP on cortisol production from cultured tumor cells and normal adrenocortical cells. 5-HT and AVP both induced a dose-dependent increase in cortisol production from cultured tumor cells. Comparison of the data obtained with tumor and normal cells, respectively, showed that 5-HT was more efficient to stimulate steroidogenesis in adenomatous than normal cells. Concurrently, the efficacy and potency of AVP were both higher in tumor than normal cells. Collectively, these results show that the abnormal in vivo responses of the adrenocortical adenoma to cisapride and LVP could be ascribed to an increased sensitivity of the tumor tissue to 5-HT and AVP. The data also suggest that the adrenocortical tumor overexpressed eutopic 5-HT4 and V1 receptors. PMID- 12530700 TI - The short synacthen test in the evaluation of adrenal masses in patients with malignancies. AB - Adrenal computed tomography with determination of Hounsfield units has proved to be sensitive and specific in the differential diagnosis of benign vs. malign adrenal lesions. On the other hand, computed tomography may fail in patients with small adrenal masses of less than 1.0 cm. However, especially in patients with diagnosed malignancies and small adrenal masses which were discovered during the diagnostic staging procedure it is important to determine the origin of the adrenal lesion. An augmented increase in 17alpha-hydroxyprogesterone (17-OHP) levels following corticotropin (1-24) stimulation has been noted in incidentally discovered adrenal masses by several groups. Therefore, we tested the hypothesis that elevated ACTH-stimulated 17-OHP (delta > 2.6 ng/mL) can predict primary adrenal lesions. We evaluated the use of the ACTH test in 85 patients with adrenocortical tumors and in 16 patients who underwent abdominal imaging for staging of a carcinoma other than of adrenal origin. We found an augmented 17-OHP response in 70 (>82%) of patients with known adrenocortical tumors and in 10 (>62%) of patients with adrenal masses and diagnosed malignancies. Results in the latter group have been confirmed in histological studies after operation or puncture. In the group of patients who suffered from a solid malignant tumor and had an adrenal mass, it was thus possible to separate primary from secondary adrenal lesions in 100%. In the group of patients with known adrenocortical tumors, it failed to differentiate between benign and malignant adrenocortical lesion in one case. We therefore think that the ACTH test is a valuable biochemical tool to distinguish primary adrenal tumors from adrenal metastasis derived from other malignancies. PMID- 12530701 TI - When fit indices and residuals are incompatible. AB - Standard chi-square-based fit indices for factor analysis and related models have a little known property: They are more sensitive to misfit when unique variances are small than when they are large. Consequently, very small correlation residuals indicating excellent fit can be accompanied by indications of bad fit by the fit indices when unique variances are small. An empirical example of this incompatibility between residuals and fit indices is provided. For illustrative purposes, an artificial example is provided that yields exactly the same correlation residuals as the empirical example but has larger unique variances. For this example, the fit indices indicate excellent fit. A theoretical explanation for this phenomenon is provided using relationships between unique variances and eigenvalues of the fitted correlation matrix. PMID- 12530702 TI - Mediation in experimental and nonexperimental studies: new procedures and recommendations. AB - Mediation is said to occur when a causal effect of some variable X on an outcome Y is explained by some intervening variable M. The authors recommend that with small to moderate samples, bootstrap methods (B. Efron & R. Tibshirani, 1993) be used to assess mediation. Bootstrap tests are powerful because they detect that the sampling distribution of the mediated effect is skewed away from 0. They argue that R. M. Baron and D. A. Kenny's (1986) recommendation of first testing the X --> Y association for statistical significance should not be a requirement when there is a priori belief that the effect size is small or suppression is a possibility. Empirical examples and computer setups for bootstrap analyses are provided. PMID- 12530703 TI - Analysis of multivariate frequency data by graphical models and generalizations of the multidimensional row-column association model. AB - Models used to analyze cross-classifications of counts from psychological experiments must represent associations between multiple discrete variables and take into account attributes of stimuli, experimental conditions, or characteristics of subjects. The models must also lend themselves to psychological interpretations about underlying structures mediating the relationship between stimuli and responses. To meet these needs, the author extends the graphical latent variable models for nominal and/or ordinal data proposed by C. J. Anderson and J. K. Vermunt (2000) to situations in which dependencies between observed variables are not fully accounted for by the latent variables. The graphical models provide a unified framework for studying multivariate associations that include log-linear models and log-multiplicative association models as special cases. PMID- 12530704 TI - Identifying criterion-related patterns of predictor scores using multiple regression. AB - Along with examples involving vocational interests and mathematics achievement, the authors describe a multiple regression based, pattern recognition procedure that can be used to identify a pattern of predictor scores associated with high scores on a criterion variable. This pattern is called the criterion pattern. After the criterion pattern has been identified, a second regression procedure can be used to estimate the proportion of variation attributable to the criterion pattern. Cross-validation can then be used to estimate the variation attributable to a criterion pattern derived from regression weights estimated in another sample. Finally, issues of criterion pattern invariance and interpretation are discussed. PMID- 12530705 TI - Comparing several robust tests of stochastic equality with ordinally scaled variables and small to moderate sized samples. AB - In a comparison of 2 treatments, if outcome scores are denoted by X in 1 condition and by Y in the other, stochastic equality is defined as P(X < Y) = P(X > Y). Tests of stochastic equality can be affected by characteristics of the distributions being compared, such as heterogeneity of variance. Thus, various robust tests of stochastic equality have been proposed and are evaluated here using a Monte Carlo study with sample sizes ranging from 10 to 30. Three robust tests are identified that perform well in Type I error rates and power except when extremely skewed data co-occur with very small n. When tests of stochastic equality might be preferred to tests of means is also considered. PMID- 12530706 TI - Comparison of closed testing procedures for pairwise testing of means. AB - A Monte Carlo simulation was conducted to compare 9 pairwise multiple comparison procedures. Procedures were evaluated on the basis of any-pair power and all pairs power. No procedure was found to be uniformly most powerful. A modification due to A. J. Hayter (1986) of Fisher's least significant difference was found to provide the best combination of ease of use and moderately high any-pair power in most cases. Pilot or exploratory studies can expect good power results with this relatively simple procedure. The greatest all-pairs power was usually provided by 1 of 2 partition-based versions of E. Peritz's (1970) procedure. Confirmatory studies will require such complex methods but may also need larger sample sizes than have been customary in psychological research. PMID- 12530707 TI - Exophiala infection from contaminated injectable steroids prepared by a compounding pharmacy--United States, July-November 2002. AB - In the United States, pharmacists compound medications to meet unique patient drug requirements or to prepare drug products that are not available commercially. In September 2002, the North Carolina Division of Public Health (NCDPH) was notified of two cases of meningitis caused by a rare fungus in patients who had received epidural injections at outpatient pain management clinics. This report describes five cases of fungal infection associated with contaminated drugs prepared at a compounding pharmacy. Clinicians should consider the possibility of improperly compounded medications as a source of infection in patients after epidural or intra-articular injections. PMID- 12530708 TI - Outbreaks of gastroenteritis associated with noroviruses on cruise ships--United States, 2002. AB - During January 1-December 2, 2002, CDC's Vessel Sanitation Program (VSP), which conducts surveillance for acute gastroenteritis (AGE) on cruise ships with foreign itineraries sailing into U.S. ports received reports of 21 outbreaks of AGE on 17 cruise ships. Of the 21 outbreaks, nine were confirmed by laboratory analysis of stool specimens from affected persons to be associated with noroviruses, three were attributable to bacterial agents, and nine were of unknown etiology. Seven outbreaks were reported in 2001, and of these, four were confirmed to be associated with norovirus (CDC, unpublished data, 2002). This report describes five of the norovirus outbreaks that occurred during July 1 December 2, 2002, on cruise ships. PMID- 12530709 TI - Measles outbreak among internationally adopted children arriving in the United States, February-March 2001. AB - On February 16, 2001, the Texas Department of Health was notified about a child aged 10 months adopted from orphanage A in China who was taken to a Texas hospital with fever, conjunctivitis, coryza, Koplik spots, and a maculopapular rash. Measles was confirmed by serologic testing. Public health authorities in Texas notified CDC, which then collaborated with health officials in other states to contact other recently adopted children from China and their adoptive families. This report summarizes the results of multistate contact investigations that identified 14 U.S. measles cases and outlines measures taken in the United States and China to control and prevent measles transmission. PMID- 12530710 TI - Individual differences in faking integrity tests. AB - The present study assessed whether success at faking a commercially available integrity test relates to individual differences among the test takers. We administered the Reid Report, an overt integrity test, twice to a sample of college students with instructions to answer honestly on one administration and "fake good" on the other. These participants also completed a measure of general cognitive ability, the Raven Advanced Progressive Matrices. Integrity test scores were 1.3 standard deviations higher in the faking condition (p<.05). There was a weak, but significant, positive relation between general cognitive ability and faking success, calculated as the difference in scores between the honest and faked administrations of the Reid Report (r=.17, p<.05). An examination of the correlations between faking success and general cognitive ability by item type suggested that the relation is due to the items that pose hypothetical scenarios, e.g., "Should an employee be fired for stealing a few office supplies?" (r=.22, p<.05) and not the items that ask for admissions of undesirable past behaviors. e.g., "Have you ever stolen office supplies?" (r=.02, p>.05: t=2.06, p<.05) for the difference between correlations. These results suggest that general cognitive ability is indeed an individual difference relevant to success at faking an overt integrity test. PMID- 12530711 TI - Perceptions of influence on child's competence among fathers in the military context. AB - This study compared the beliefs of fathers serving in the military with those of a matched sample of civilian fathers on the time spent with their kindergarten child and their perceptions of their influence as well as their spouse's influence on their child's cognitive and emotional competence. Parents are considered prime agents of change during early childhood. However, fathers who serve in the military face challenges unique to this context and reflective of the frequent and intense demands on their time and attention. Results suggest that, in a time of peace, military and civilian fathers share similar perceptions and time reported spent with their child. Mothers were perceived to be equivalent in their influence to the military father. PMID- 12530712 TI - Environments of physically neglected children. AB - A reanalysis of data from the Gaudin, Polansky, Kilpatrick, and Shilton 1996 study of 185 low-income families combined information from neglectful families with information from the comparison group. Physical neglect was related to maternal depression, more adult problems, stressful life events, negative family affect, and low social support. Multivariate analyses indicated that a combination of higher maternal education, positive family affect, more supportive services, and fewer stressful life events predicted the provision of adequate physical care among low-income families. PMID- 12530713 TI - Correlations for leaders' anticipation of targets' resistance and targets' ratings of leaders' influence tactics: a replication. AB - A sample of 553 leader-target dyads was used to replicate the 2001 work of Barbuto, Scholl, Hickox, and Boulmetis. The study examined the correlation between leaders' anticipation of targets' resistance and influence tactics used by leaders to persuade targets. The original study reported mixed findings of various hard and soft tactics used by leaders based on the different zones of resistance anticipated. Present results show no relation between leaders' anticipation of targets' anticipated resistance and the influence tactics used. PMID- 12530714 TI - Depression among university students in South Africa. AB - Of 129 first-year students at an historically Black university students in psychology reported significantly less depression than nonpsychology students, and younger students reported significantly lower scores on depression than older students on the Beck Depression Inventory. PMID- 12530715 TI - Working memory and reference memory in adult rats following limbic seizures induced at 21 or 90 days of age. AB - Rats were either seized or not seized at 21 days of age (weaning) or at 90 days of age with a single systemic injection of lithium (3 mEq/kg) and pilocarpine (30 mg/kg). When tested as adults (120 days of age) for spatial memory in the Olton radial maze, the rats that had been seized as adults exhibited about five times more working (short-term) memory errors than the other three groups which did not differ significantly from one another. The numbers of errors for long-term (reference) memory did not differ significantly among the four groups. The deficits in working memory for the group seized as weanlings and reported previously were not replicated. One possible explanation for this discrepancy might be differential effects upon brain organization associated with seizures evoked by injecting the pilocarpine 24 hr. rather than 4 hr. after the lithium. PMID- 12530716 TI - Intercessory prayer in psychological well-being: using a multiple-baseline, across-subjects design. AB - Undergraduates from an upper-level psychology course were volunteer participants in the study. The 8 participants were to be prayed for in a Multiple Baseline Across Subjects research design, which included a 1-week minimum baseline period for all subjects followed by the sequential presentation of the independent variable so that every two weeks, two additional subjects were being prayed for until all but 2 participants, who maintained baseline, were exposed to being prayed for at 7 weeks. All participants were prayed for by one of the experimenters using a nondirective method of prayer where no specific requests were made. All subjects completed the Taylor Manifest Anxiety Scale on a daily basis for 5 weeks and the Minnesota Multiphasic Personality Inventory-2 on a weekly basis for 7 weeks. Analysis of data identified significant reductions in anxiety scores on both the tests for subjects who were prayed for but not for those who were not prayed for. Subjects' lower mean anxiety scores somewhat matched the sequential timing of being prayed for. PMID- 12530717 TI - Alcohol law changes and homicide in Iceland. AB - Homicide rates declined, but not significantly, after the passage of a law in Iceland legalizing the sale of strong beer. PMID- 12530718 TI - Sex differences in cerebral tissue volume and cognitive performance during adolescence. AB - Adolescent development is associated with progressive increases in the ratio of cerebral white-to-gray matter volume, but it is unclear how these changes relate to cognitive development and whether they are associated with sex-specific variability in cerebral maturation. We examined sex differences in the relation between cerebral tissue volume and cognitive performance in 30 healthy adolescents (ages 13 to 17 years using morphometric magnetic resonance imaging (MRI). In the 10 boys, greater white matter volume during adolescence was positively correlated with faster speed of information processing and better verbal abilities, while cerebrospinal fluid volume was negatively correlated with verbal abilities. No significant relations between cerebral tissue volume and cognitive abilities were found for the sample of 20 girls, raising the possibility of a different developmental trajectory for females that was not sampled in the age range of this study. Findings suggest sex-specific developmental differences in the relations between cerebral structure and function. PMID- 12530719 TI - National ratings of happiness, suicide, and homicide. AB - In a sample of 15 industrialized nations, mean ratings of happiness were not significantly associated with suicide or homicide rates. PMID- 12530720 TI - Moral judgment and drivers' behavior among Brazilian students. AB - It is well known that lifestyle, social deviance, and criminal background are related to involvement in road accidents. This study addressed the connection between moral judgment and driving behavior, especially deliberate violations of the traffic law. 260 university students (age 18-30 years, 41% women) completed the Sociomoral Reflection Measure and Driver Behavior Questionnaire. No significant associations were found, probably due to the small variance in Moral Judgment score in this selected sample. This study presented a Portuguese 28-item version of the Driver Behavior Questionnaire that reproduced the same four-factor structure as the British version: ordinary violations, aggressive violations, errors, and lapses. PMID- 12530721 TI - Sex differences in the relationship between interpersonal problems and defensive style. AB - Horowitz, et al.'s Inventory of Interpersonal Problems and Bond, et al.'s Defensive Style Questionnaire were completed by 42 women and 35 men. Significant correlations emerged between most interpersonal problems and the tendency for both men and women to use immature and intermediate defense mechanisms in both work and intimate relationships. However, women were more likely than men to employ immature defenses when dealing with issues of "control" in intimate relationships while men were more likely to employ intermediate defenses in response to problems with "intimacy" in work relationships. Data support further inquiry into sex differences in interpersonal problems and defensive style. PMID- 12530722 TI - Family contexts, individual characteristics, proximal settings, and adolescents' aspirations. AB - This study examined relations among family contexts, individual characteristics, proximal family and school settings, and adolescents' aspirations. As part of a national longitudinal investigation of Australian secondary school students, data were collected from 1,724 boys and 1,788 girls. The results suggest (a) family contexts defined conjointly by family social status and parents' perceived aspirations have medium to large associations with adolescents' educational aspirations and small significant associations with occupational aspirations. (b) Also, relations between family contexts and adolescents' aspirations are mediated in part by measures of adolescents' individual characteristics and their perceptions of proximal learning settings. (c) There are family context differences in the nature of the linear and curvilinear relations among individual characteristics, proximal settings, and adolescents' aspirations. Further, (d) there are sex-related variations in the associations between family contexts, individual characteristics, proximal learning settings, and adolescents' aspirations. PMID- 12530723 TI - Mortality from abortion after Roe vs Wade. AB - The decline in mortality from abortions after Roe vs Wade was probably a result of the introduction of safer procedures for abortions, but the decline in mortality was greater for induced abortions than for other types of abortions. PMID- 12530724 TI - Association of perceived parental attitudes towards premarital sex with initiation of sexual intercourse in adolescence. AB - To assess the association between perceived parental disapproval of premarital sex, quality of the parent-daughter relationship, and initiation of sexual intercourse, a sample of 614 economically deprived female students in Chile were tested. The perception of disapproval of premarital sex by the mother along with a good mother-daughter relationship appears to constitute a buffer against early initiation of sexual activity in adolescence. Results are consistent with findings reported in studies conducted in the USA. PMID- 12530725 TI - Spirituality and complex partial epileptic-like signs. AB - Using a sample of 262 undergraduate volunteers, correlational and regressional analyses were completed to examine the relation of self-reported complex partial epileptic-like signs to a five-dimensional model of spirituality assessed by the Expressions of Spirituality Inventory. Analyses show that spirituality is significantly predictive of Complex Partial Epileptic-like Signs. Scores from subscales Paranormal Beliefs, Experiential/Phenomenological Dimension, and Existential Well-being uniquely accounted for significant portions of Complex Partial Epileptic-like Signs variance. ESI Cognitive Orientation Toward Spirituality significantly correlated with Complex Partial Epileptic-like Signs but it was not a significant predictor in regression analyses. ESI Religiousness was unrelated to Complex Partial Epileptic-like Signs in all analyses. This pattern of relations remained largely the same when participants' age, sex, and reported religious involvement were controlled. The paper includes a discussion of the meaning of the findings and suggestions for research. PMID- 12530726 TI - Participants' ratings of male physicians who vary in race and communication style. AB - Research has shown minorities receive lower quality health care than White persons even with socioeconomic conditions controlled. This difference may partially be related to racially biased attitudes and impaired communication in interracial relationships between physicians and patients. This study investigated the effect of physicians' race and nonverbal communication style on participants' evaluations. Patients at a local health clinic were participants (N = 116: 84% Black, 16% White). Each participant viewed one of four videotapes showing varied race of a physician (Black or White) and the physician's nonverbal behavior (expressing concern or distance), and then completed a questionnaire evaluating the depicted physician. Overall, participants did not give significantly different preferences for physicians of the same race. However, participants' evaluations were significantly associated with physicians' nonverbal style. Nonverbal concern was associated with highest satisfaction, trust, self-disclosure, recall of information, likelihood of recommending the physician, and intent to comply with the physician's recommendations. When male and female participants were compared, preference for a physician of the same race was found only among male participants who viewed verbally distant physicians. Results suggest that social skills are more important than race in shaping patients' perceptions of physicians. PMID- 12530727 TI - Relations among self-talk, self-consciousness, and self-knowledge. AB - The aim of this study was to examine the relations among self-talk, self consciousness, and self-knowledge through an exploratory principal component analysis and to test the hypothesis that only the functional and reflective aspects of self-consciousness contribute to self-knowledge. A self-report questionnaire including 6 scales assessing different aspects of self-talk, self consciousness, and self-knowledge was administered to 203 German undergraduate university students. A principal component analysis of the scales yielded a two factor solution, supporting the distinction between functional and dysfunctional self-consciousness. In a stepwise multiple regression analysis, only functional self-consciousness was a significant predictor of self-knowledge. Limitations of the present measures of inner speech are addressed. PMID- 12530728 TI - Identification and remediation of reading difficulties based on successive processing deficits and delay in general reading. AB - Widespread learning problems among South African children are associated with the apartheid era and show a need for effective reading programs. In selecting these programs, it is useful to differentiate between children with dyslexia and children whose reading is poor because teaching was inadequate. In this study, the Woodcock Tests of Reading Mastery-Revised and tests modelled on the Cognitive Assessment System were used to define a group of children with deficits in successive processing associated with dyslexia and a group of children with general reading delay. There were two girls and five boys in each group. For the children with successive processing deficit, the mean age was 9 yr., 8 mo. For the other group, mean age was 9 yr., 3 mo. Control groups were matched for age and sex and kind of reading difficulty. The first group received Das's PASS Reading Enhancement Program, and the second participated in a remedial program based on Whole Language principles. The treatment groups received 24 1-hr. long sessions. Gains in successive processing were shown for the first group, as measured by the tests modelled on Cognitive Assessment System subtests but not for the second group. Both groups showed gains in phonics and word identification, relative to their respective control groups, suggesting the respective intervention program was effective for each group. PMID- 12530729 TI - Significant effects due to rephrasing the Maslach Burnout Inventory's personal accomplishment items. AB - Subjects, 292 Dutch psychology students answering negatively phrased Personal Accomplishment items, reported a more personally competent feeling than subjects answering positively phrased personal accomplishment items. The way of phrasing personal accomplishment items significantly affects the answers given. PMID- 12530730 TI - Construct validity of the Piers-Harris Children's Self-concept Scale. AB - The aim of the present study was to investigate the construct validity of the Piers-Harris Children's Self-concept Scale with a group of 511 (264 girls, 247 boys aged 9 to 12 years) Greek children. Principal Components Analysis was performed. Six factors emerged, i.e., Behavior, Anxiety, Intellectual and School Status, Popularity, Physical Appearance and Attributes, and Self-contentment. PMID- 12530731 TI - Positive health in a group of Swedish white-collar workers. AB - Ryff's Psychological Well-being scales cover six dimensions of psychological well being (Self-acceptance, Environmental mastery, Positive relations with others, Personal growth, Purpose in life, and Autonomy) and have been suggested as an adequate measure of positive psychological functioning. Apart from translating the scales to Swedish and examining the psychometric properties of the measure, the present study aimed to explore the relationships between the Ryff scales and the General Health Questionnaire, negative affectivity, and physical symptoms using self-ratings from 91 full-time employed women and men. Given low internal consistency for the different dimensions of the Ryff scales, correlational analyses were based on a composite index. Analysis indicated negative relations between the Ryff index and other measures and are in line with prior findings showing that the index taps positive psychological functioning while other indices focus on negative functioning. PMID- 12530732 TI - Daily variations in cortisol levels and binge eating disorder. AB - Morning and afternoon levels of cortisol for 73 volunteers (67 women and 6 men) were compared in relation to their Binge Eating Disorder scores, Body Mass Indexes, and self-reports of mood and hunger. Cortisol level was not significantly correlated with binge eating or mood or hunger for either time period. However, it was inversely related to body mass, with lower cortisol levels associated with greater body mass. PMID- 12530734 TI - Scores on liking and solidarity in interpersonal relationships. AB - This study examined liking and solidarity in interpersonal relationships. Participants were 226 undergraduates (89 men, 137 women: M age=27.0) who identified (by initials) an individual with whom they had a recent interaction and referenced that interaction when completing Rubin's Liking Scale and Wheeless's Interpersonal Solidarity Scale. A correlation of .73 (p<.0001) was obtained between the scores on these scales, suggesting when individuals like one another, they report feelings of solidarity also. PMID- 12530733 TI - Perceived abuse and disability in a sample of Ohio's women's correctional population. AB - The Abuse Disability Questionnaire was administered to 435 female inmates at a state prison in Ohio. Analysis indicated that 69% of the women reported some type of past physical, sexual, or psychological abuse, which is consistent with other reports for state prisons. Both the amount of abuse reported as well as the extent of associated psychological impairment was less in comparison to scores for women in domestic violence shelters. While all women in the prison were screened, those who met criteria for psychiatric diagnoses had significantly higher Abuse Disability Questionnaire scores than those who were not so diagnosed by a mental health professional. As in prior studies with the questionnaire, a significant relation between reported exposure to prior abuse and perceived impairment was found. PMID- 12530735 TI - Positive thinking in bereavement: is it related to depression, anxiety, or grief symptomatology? AB - The present study sought to explore the relationship between positive thinking- operationalized as the frequency of positive automatic thoughts and dispositional optimism--and depression, anxiety, and traumatic grief symptomatology. Data were derived from 326 bereaved adults who had 3 yr. earlier experienced the loss of a first degree loved one. Analysis showed that positive thinking was inversely related to measures of depression, anxiety, and traumatic grief symptomatology. However, positive thinking did not covary with anxiety and traumatic grief when shared variance was controlled. Conversely, positive thinking remained inversely related to depression, even when anxiety and traumatic grief were controlled. The findings indicate that, among bereaved individuals, low positive thinking is a cognitive feature of depression that is relatively independent of anxiety and traumatic grief symptomatology. PMID- 12530736 TI - Suicide among murderers: England and Wales 1901 to 1949. AB - From 1901 to 1949 in England and Wales, the suicide rate of murderers was associated with the overall English suicide rate, and both rates had similar social correlates. PMID- 12530737 TI - Reliability and factor structure of the Brazilian version of the Center for Epidemiologic Studies-Depression. AB - The Center for Epidemiologic Studies-Depression scale is a screening test designed by the National Institute of Mental Health (USA) to screen for 'depressed' individuals in the community. It has been widely used in different settings and cultures displaying variable psychometric properties. The present article describes coefficients of internal consistency and factor analysis of the CES-Depression version adapted for use in Brazil. In a sample of 523 college students internal consistency measured by Cronbach alpha was .85. Structure analysis evidenced that two questions were poorly correlated with the remaining questions. Factor analysis resulted in a four-dimension solution accounting for 53.8% of the total item variance. The scale is adequate for use in this different cultural setting. PMID- 12530738 TI - Young children's participation in everyday family and community activity. AB - Patterns of participation in everyday family and community activities are described for children from birth to 6 years of age. Parents or other primary caregivers completed a survey of either family life or community life as sources of children's learning opportunities and experiences. Rates of children's participation in 50 family activities and 50 community activities at different ages were identified and analyzed in terms of similarities and differences in participation patterns. Findings indicated considerable variability in involvement in 100 family and community activities by young children of different ages. Implications for using everyday family and community activities as sources of enriched learning opportunities that enhance children's development are discussed. PMID- 12530739 TI - Decriminalization of suicide in seven nations and suicide rates. AB - In a study of seven nations, suicide rates were higher in the years after decriminalization of suicide than before. PMID- 12530740 TI - Hindsight and confirmation biases in an exercise in telepathy. AB - Belief in the paranormal or claims of paranormal experiences may be, at least in part, associated with systematic cognitive biases. 48 undergraduate college students engaged in an exercise in telepathy in which the color of cards was 'sent' to them by the experimenter under two conditions. In a Hindsight-possible condition, participants recorded whether their choice was correct following the revelation of the color. In the Control condition participants committed to a particular response by writing it down before receiving feedback, thus eliminating ability to alter retrospectively what 'was known all along'. Consistent with a hindsight bias, participants performed significantly better under the Hindsight-possible condition. Moreover, a statisically significant correlation was found between paranormal belief assessed on Tobacyk's 1988 Revised Paranormal Belief Scale in the Hindsight-possible but not in the Control condition, suggesting a confirmation bias. Results are discussed in terms of interactions between hindsight and confirmation biases and how they might relate to paranormal beliefs. PMID- 12530741 TI - Overload and work-family conflict among Australian dual-career families: moderating effects of support. AB - Individuals in dual-career situations have become increasingly common all over the world. For couples dealing with multiple demands, this lifestyle often generates stresses and strains at home and at work, which can have negative consequences for organizations. Most empirical research into this lifestyle has been conducted in the United States and Britain, and very little in Australia. This particular study, based on data from an Australian sample of 65 dual-career couples, analyzed the relation between overload and work-family conflict and the moderating effects of support (supervisor, coworkers, and friends). Results confirm that overload was significantly related to work-family conflict but no moderating effects were found for support. Limitations of the study and an organizational role in managing the work-family interface dual-career couples are discussed. PMID- 12530742 TI - Clustering in rampage murders. AB - A study of 81 incidents of rampage murders in the USA from 1988 to 1999 yielded no evidence for clustering. PMID- 12530743 TI - Tennessee Self-concept Scale scores of urban African-American women. AB - The Tennessee Self-concept Scale: Second Edition scores of 33 urban, African American women were compared in a pretest-posttest design. The treatment was exposure to a 3-mo. literacy and social skills training program. Contrary to expectations, scores on just two subscales--Self-criticism and Physical Self- fell outside the average range. Also, Physical Self was the only scale score to change significantly (-3.83). These results suggest that self-esteem scores of urban African Americans were similar to those of women in the general population. PMID- 12530744 TI - Academic elite in social work: 1994-2000. AB - The 11 most highly rated graduate programs in social work in 1994 maintained their positions in 2000. These "elite" programs showed little or no change in their hiring of faculty from among their own and one another's graduates. PMID- 12530745 TI - Examination of relations between the NEO Personality Inventory-Revised and the Temperament and Character Inventory. AB - The present investigation examined the relation of the Five Factor Model of personality as measured by the NEO Personality Inventory-Revised to the seven factor model of temperament and character as tapped by the Temperament and Character Inventory by testing a sample of 344 undergraduate students. Correlations and regression analyses suggest there is substantial overlap between the NEO-Personality Inventory-Revised domains and Temperament and Character Inventory dimensions as reflected in multiple correlations ranging from .54 to .80 for the latter when used as predictors of NEO Personality Inventory-Revised domains and .46 to .78 for the former domains when used as predictors of Temperament and Character Inventory dimension scores. Information for constructing the regression equations to allow for the prediction of personality constructs for one model from scores obtained on the other are provided. PMID- 12530746 TI - Thinking styles of school teachers and university students in mathematics. AB - This research studied the thinking styles of 18 school teachers and 15 college students in the field of mathematics, using the Inquiry Mode Questionnaire by Harrison and Bramson. The multivariate analysis of variance showed that the 21 female teachers and college students scored as more Idealistic than their 12 male peers. There also was a significant group-by-sex interaction, which indicated that the female college students preferred the Analyst thinking style more frequently than their male peers, whereas the male teachers preferred the Analyst style more frequently than the female teachers. On the whole, the most preferred thinking style was the Analyst style, and the least preferred one was the Synthesist style. The implications of findings are discussed. PMID- 12530747 TI - Self-efficacy in a sample of education majors and teachers. AB - The teachers' self-efficacy, both personal and general, has a profound effect on students' learning. This study investigated the influence on education students' perceptions of their experience as teaching interns of Personal Teaching Efficacy and General Teaching Efficacy. The participants were 196 undergraduates and graduate students who were preparing for or active as interns in teaching or were teachers. There was a significant effect between Personal Teaching Efficacy and General Teaching Efficacy scores, with all respondents scoring higher on the former scale. A significant interaction indicated that students who had not completed internships scored significantly higher on General Teaching Efficacy than either students who had just completed internships or those who were engaged in teaching. Implications are discussed. PMID- 12530748 TI - Comment on "Convergent validity of the Collett-Lester Fear of Death and Templer Death Anxiety Scales in Egyptian male college students". AB - Correlations for scores on Templer's Death Anxiety Scale and the Collett-Lester Fear of Death Scale indicate a lack of validity for Templer's scale. PMID- 12530749 TI - Aural, visual, and pictorial stimulus formats in false recall. AB - The present investigation is an initial simultaneous examination of the influence of three stimulus formats on false memories. Several pilot tests were conducted to develop new category associate stimulus lists. 73 women and 26 men (M age=21.1 yr.) were in one of three conditions: they either heard words, were shown words, or were shown pictures highly related to critical nonpresented items. As expected, recall of critical nonpresented stimuli was significantly greater for aural lists than for visually presented words and pictorial images. These findings demonstrate that the accuracy of memory is influenced by the format of the information encoded. PMID- 12530750 TI - Ethics in family vs nonfamily owned businesses. AB - This study investigated the ethical judgments in family vs nonfamily owned businesses. Respondents rated their ethical evaluations of 5 scenarios which involved ethical issues, e.g., promotion of a family member ahead of an equally qualified nonfamily employee. Analysis of responses from 120 participants yielded no statistically significant differences between the ethical judgment ratings of owner/managers of family and nonfamily owned firms. PMID- 12530751 TI - Least sum of Euclidean regression residuals: estimation of effect size. AB - A universal measure of the magnitude of effect size is introduced for Euclidean based regression analyses of experimental designs. The measure is based on the ratio of the sums of Euclidean residuals obtained under the alternative and null hypotheses. PMID- 12530752 TI - Possible influence of defenses and negative life events on patients with chronic fatigue syndrome: a pilot study. AB - 13 patients with a diagnosis of chronic fatigue syndrome and two contrast groups of conversion disorder patients (n = 19) and healthy controls (n = 13) were assessed using the projective perceptual Defense Mechanism Test to investigate if specific defense patterns are associated with chronic fatigue syndrome. Another objective was to assess the possible influence of perceived negative life events prior the onset of the illness. The overall results showed significant differences in defensive strategies among groups represented by two significant dimensions in a Partial Least Squares analysis. Compared to the contrast groups the patients with chronic fatigue syndrome were distinguished by a defense pattern of different distortions of aggressive affect, induced by an interpersonal anxiety-provoking stimulus picture with short exposures. Their responses suggested the conversion group was characterized by a nonemotionally adapted pattern and specific constellations of defenses, associated with interior reality orientation compared to the patients with chronic fatigue syndrome and the healthy controls. Rated retrospectively, the group with chronic fatigue syndrome reported significantly more negative life events prior to the onset of their illness than healthy controls. For instance, 5 of the 13 patients reported sexual assault or physical battery as children or teenagers compared to none of the healthy controls. A significant association was found between defense pattern and frequency of reported negative life events. However, these retrospective reports might be confounded to some extent by the experience of the patients' illness; for example, the reports may be interpreted in terms of present negative affect. PMID- 12530753 TI - Behavior Evaluation Scale: validity in studies on social anxiety. AB - The present study was an initial effort to evaluate the use of the Behavior Evaluation Scale, a measure of the tendency to perceive behavior as pathological, in studies on social anxiety. 61 undergraduate students provided ratings on attributional dimensions and completed the scale as well as the Fear of Negative Evaluation questionnaire. There was a moderate correlation of .48 between the Behavior Evaluation Scale scores and fear of evaluation. In addition, participants who perceived the behavior as pathological scored higher on fear of evaluation only if they made internal or general attributions. Results suggest that the perception of behavior as measured by the Behavior Evaluation Scale and the fear of evaluation are distinct constructs. Suggestions for further examination of the scale's validity are discussed. PMID- 12530754 TI - Students' course and faculty evaluations: an informal look. AB - For one class, over 8 consecutive semesters, about 10% of the students completing a course and faculty evaluation form reported one or more session cancellations, while in actuality there were none. PMID- 12530755 TI - Influences of type of high school attended and current relationship status on life goal ratings of college women. AB - Scores on the Goals in Life Scale of 183 college women were compared based on type of high school attended, single-sex versus co-educational, and current relationship status, dating versus nondating. Type of high school alone did not influence any life goal, and those in dating relationships gave priority to relationships over life goals. Several significant interaction effects were present, however, as single-sex school attendees in dating relationships gave lower ratings to several life goals than did women in the other groups. PMID- 12530756 TI - Internal consistency of the Yamauchi/Templer Money Attitude Scale. AB - The factor pattern of Yamauchi and Templer's Money Attitude Scale was replicated with a sample of 72 undergraduates. PMID- 12530757 TI - Psychometric properties of the Francis Scale of Attitude Towards Christianity among Portuguese university students. AB - To facilitate use of the adult form of the Francis Scale of Attitude Towards Christianity in cross-cultural studies, the psychometric characteristics of the translated scale were examined among 323 university students in Portugal (130 men and 193 women). Their ages ranged from 18 to 31 years. Analysis supported the unidimensionality, internal consistency, and construct validity of this scale in this sample of Portuguese university students. PMID- 12530758 TI - Effects of brief training on application of the Global Assessment of Functioning Scale. AB - 31 staff members within one VA medical center were briefly trained in using the Global Assessment of Functioning scale. At both pre- and posttraining participants provided Global Assessment of Functioning scores and identified their GAF-derivation strategies for two vignettes describing hypothetical patients. Analysis showed that initially there was inter- and intrarater inconsistency in strategy for deriving scores and that training increased consistency. Participants persisted in giving ratings that exceeded the criterion score. However, training significantly reduced Global Assessment of Functioning scores for one vignette, bringing scores closer to the criterion, and a significant number of people gave lower ratings after training. The study highlights common errors and points to the need for formal training in use of the scale. PMID- 12530759 TI - Individual personality characteristics related to suggestibility. AB - The current study investigated the relationship between suggestibility of memory, personality characteristics identified by the Millon Index of Personality Traits, and tolerance for ambiguity measured by MacDonald's Ambiguity Tolerance-20. 85 female and 16 male college students were assigned to either an experimental group receiving the suggestive information or a control group. Suggestibility was assessed using Lindberg's suggestibility measure consisting of a short video, followed by a questionnaire used to assess memory, and a second administration one week later. Logistical regression analyses were used to construct a model of the personality characteristics predictive of suggestibility and indicated that susceptibility to suggestive information may differ across personalities for variables such as sensing, innovating, agreeing, and low tolerance of ambiguity. PMID- 12530760 TI - Relation of test-specific motivation and anxiety to test performance. AB - The relations between consequence of test scores and motivation, anxiety, and test performance were studied with 112 persons in four undergraduate educational psychology courses. Students were given two versions of an hourly course examination that varied in consequence, with one counting for part of their grade and the other not counting. Each student completed the Learning and Study Strategies Inventory prior to taking the examination. Afterwards students completed a measure of test anxiety and test motivation specific to the examination they had just taken. Significant main effects were found for consequence of test with scores for test anxiety, test performance, and test motivation. Also, the subscales showed a consistent pattern of relations with test performance and test anxiety across the two conditions, but not for test motivation for which few relations were found under the condition with no consequence. PMID- 12530761 TI - A study of intelligence in Estonia. AB - The Standard Progressive Matrices was standardised in Estonia in 2001 on a sample of 2,689 12- to 18-yr.-olds. The mean IQ of the Estonian sample was approximately 5 points higher than British IQ norms of 1979. Adjustment for the estimated secular increase of intelligence in Britain reduces the mean IQ of the Estonian sample to 100.2, in relation to a British mean of 100. A particular interest of the result from Estonia is that the intelligence of a European population is apparently not impaired by decades of low living standards. PMID- 12530762 TI - Lower incidence of the upper respiratory tract infections among general practitioners as compared to their patients. AB - A retrospective cohort study suggest that general practitioners (GPs) run a significant reduced risk (more then 50%) to develop an upper respiratory tract infection (URTI) with fever compared to their patients. This difference can't be explained by variables such as gender, age, children and particularly young children, vaccination against influenza, presence of chronic illness or allergies, medication taken on a regular basis, smoking habits and regular physical exercise. PMID- 12530763 TI - Smoking cessation or reduction in women attempting to conceive after ectopic pregnancy. AB - The relationship between smoking cessation or reduction and attempt to conceive was studied in 646 women treated for ectopic pregnancy (EP) (Auvergne register of EP, France). Women were prospectively followed. Tobacco consumption was recorded as the mean number of cigarettes smoked per day at the time of interview. Smoking reduction was defined as stopping smoking or decreasing tobacco consumption by at least 3 cigarettes per day. Statistical analysis was performed using conditional logistic regression. The results confirmed that women reduced tobacco consumption when they became pregnant. However, no association was found between smoking reduction and attempting to conceive again. These results suggest that both women and physicians need to be informed about the possible tobacco consumption effects of smoking on fertility. PMID- 12530765 TI - Validation of general practitioner-diagnosed COPD in the UK General Practice Research Database. AB - BACKGROUND: Information in large, automated databases can be useful to study the natural history of respiratory diseases in the community, but the validity of definitions needs to be demonstrated. AIM: To compare a simple computer algorithm that identifies patients diagnosed with chronic obstructive pulmonary disease (COPD) and severity of COPD in the UK General Practice Research Database (GPRD) with general practitioner (GP) clinical records, to evaluate the utility of this algorithm for identifying COPD patients and for distinguishing COPD from asthma. METHODS: Using a computer algorithm identifying patients by diagnostic codes and allotting three grades of severity by drug use, a sample of 225 patients in the GPRD with a diagnosis of COPD and an age-sex matched group of 75 patients with asthma were randomly selected. Questionnaires were posted to the GPs of the 300 selected patients who were asked to state diagnosis and to grade severity based on the individual's medical record. Agreement was quantified with the kappa index, an estimator that accounts for agreement that occurs by chance. RESULTS: Response rate was 85.7%. The concordance between COPD diagnosis by the GPRD algorithm with that of the GP was quantified as a kappa of 0.52, and the concordance between COPD severity by the GPRD algorithm with that of the GP was quantified as a kappa of 0.54. The kappa index for COPD diagnosis increased with increasing severity of COPD (0.46, 0.59, and 0.68 for mild, moderate and severe COPD, respectively), but similar good agreement was observed in a stratified analysis by sex, age, smoking status and number of comorbidities. CONCLUSIONS: It is concluded that the GPRD algorithms used for diagnosis and severity of COPD are a good screening tool for COPD in the UK general population, and satisfactorily differentiate COPD from asthma patients, particularly when disease is moderate or severe. PMID- 12530764 TI - Detection and molecular typing of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks and in different patient samples from southwest Germany. AB - The prevalence of different genospecies of Borrelia burgdorferi sensu lato in infected ticks could be a determinant for the risk of acquiring Lyme borreliosis (LB) and its clinical presentation. A total of 7373 ticks and 2761 samples from LB patients from the same area in southwest Germany were analyzed by PCR to assess the frequency of the occurrence of LB-associated genospecies. Fifteen percent of the tick samples and 19% of the human samples were found positive for the presence of B. burgdorferi sensu lato. Further identification of 1106 B. burgdorferi sensu lato positive tick samples by reverse line blotting and 125 positive patient samples by nested PCR using species-specific primers revealed the occurrence of B. afzelii, B. burgdorferi sensu stricto, B. garinii and B. valaisiana. Both single-species and mixed infections were noted and a similar distribution of the different genospecies was found in ticks compared with human samples. It was also the purpose of this study to obtain more information about a possible correlation between the distribution of Borrelia species and clinical syndromes of LB. Skin biopsies of 59 patients with acrodermatitis chronica atrophicans and cerebrospinal fluid samples from 78 patients with possible neuroborreliosis were analyzed. In conclusion, the distribution of the different genospecies in ticks is the decisive factor for the occurrence of the different Borrelia genospecies in samples from LB patients. Borrelia afzelii is the predominant genospecies in all kind of samples from the observed area and there seems to be no association of particular Borrelia genospecies with distinct clinical manifestations of LB. PMID- 12530766 TI - Associations between infections and fatigue in a Dutch working population: results of the Maastricht Cohort Study on Fatigue at Work. AB - The relationship between fatigue and common infections was further explored, as part of a 3 year prospective cohort study on Fatigue at Work. The current study is based on seven successive questionnaires, covering the first 2 years of follow up. The overall response at baseline was 45% (n = 12,140). On T1 10,592 (87.2% compared to baseline response) employees returned the questionnaire. For T2, T3, T4, T5 and T6, 10,270 (84.6%), 9655 (79.5%), 8956 (73.8%), 8692 (71.6%) and 8070 (66.5%) employees respectively returned the questionnaire. Self-administered questionnaires were used to determine the level of fatigue with the Checklist Individual Strength (CIS) and the occurrence of common cold, flu-like illness and gastroenteritis. Regression analysis using generalized estimated equations (GEE) were used for data analysis. We found a cross-sectional relationship between fatigue and the infections flu-like illness and gastroenteritis, and a longitudinal relationship between an infection as a predictor of fatigue. For fatigue as a predictor of an infection, we found odds ratios (ORs) of 1.35 (confidence interval (CI) 1.28-1.42) for flu-like illness and 1.33 (CI: 1.25 1.42) for gastroenteritis. The highest incidence of infections was found among employees who reported high fatigue levels on two successive occasions. The increased incidence of infections, is regarded as a substantial effect of fatigue because it is associated with significant absenteeism from work and leads to discomfort. PMID- 12530767 TI - Hypertension and combinations of cardiovascular risk factors. An epidemiologic case-control study in an adult population in Guadeloupe (FWI). AB - Our aim was to quantify the association between hypertension and four well-known cardiovascular risk factors (obesity, dyslipidemia, high blood glucose, smoking) and to determine the extent of their combinations in hypertensive subjects in an adult population in Guadeloupe (FWI). A case-control study was conducted in 1999. Odds ratios (ORs) and 95% confidence interval (95% CI) of hypertension according to cardiovascular risk factors were calculated by a logistic regression model. In all, 4210 subjects were included in the study: 2105 hypertensives and 2105 age and sex matched non-hypertensive controls. In the hypertensives, obesity (30%) was the most frequent risk factor followed by dyslipidemia (23.2%), current smoking (11.5%) and high blood glucose (8.2%). But, prevalence of current smokers was higher in normotensives (13.9 vs. 11.5%, p < 0.01). Among cases, 44.5% had no risk factor other than hypertension. The proportion of subjects with multiple risks factors (< or = 2) was higher in hypertensives than in controls (55.5 vs. 6.5%,p < 0.001) and higher in women than in men in cases (58.1 vs. 51.8%, p = 0.005). The adjusted ORs (95% CI) of hypertension were 2.41 (2.02-2.88) for obesity, 1.39 (1.16-1.66) for dyslipidemia and 1.67 (1.20-2.32) for high blood glucose. Our study documents the high prevalence of combinations of cardiovascular risk factors in hypertensive subjects in Guadeloupe and confirms the need of a global risk approach in prevention and treatment of hypertension. PMID- 12530769 TI - Infectious diseases before and after German unification: trends in mortality and morbidity. AB - After the fall of the Berlin Wall in 1989 the former communist countries of central and eastern Europe underwent substantial social and economic changes that had a major impact on health. The situation of the former German Democratic Republic was unique in that its existing institutions were replaced rapidly and it was quickly integrated economically with the West. This study describes recent trends in morbidity and mortality from infectious diseases in East and West Germany before and after unification using routine data from 1980 onwards. Substantial differences in morbidity and mortality resulting from infectious diseases were observed between East and West Germany. These seem to be the complex result of societal and health system differences in both parts of Germany before unification, differing population dynamics and health behaviours, and an increasing westernisation in the eastern part of Germany during recent years. PMID- 12530768 TI - Predictors of stroke mortality in elderly people from the general population. The CArdiovascular STudy in the ELderly. AB - Stroke occurs particularly frequently in elderly people and, being more often disabling than fatal, entails a high social burden. The predictors of stroke mortality have been identified in 3282 subjects aged > or = 65 years, taking part in the CArdiovascular STudy in the ELderly (CASTEL), a population-based study performed in Northeast Italy. Historical and clinical data, blood tests and 14 year fatal events were recorded. Continuous items were divided into quintiles and, for each quintile, adjusted relative risk (RR) with 95% confidence intervals [CI] was derived from multivariate Cox analysis. Age, historical stroke (RR: 5.2; 95% CI: 3.18-8.6) and coronary artery disease (RR: 1.38; CI: 1.18-2.1), atrial fibrillation (RR: 2.40; CI: 1.42-4.0), arterial hypertension (RR: 1.33; CI: 1.15 1.76), systolic blood pressure > or = 163 mmHg (RR: 1.84; CI: 1.20-2.59), pulse pressure > or = 74 mmHg (RR: 1.50; CI: 1.13-2.40), cigarette smoking (RR: 1.60; CI: 1.03-2.47), electrocardiographic left ventricular hypertrophy (RR: 1.72; CI: 1.10-2.61), impaired glucose tolerance (IGT, RR: 1.83; CI: 1.10-3.0), uric acid (UA) > 0.38 mmol/l (RR: 1.61; CI: 1.14-2.10), serum potassium > or = 5 mEq/l (RR: 1.70; CI: 1.24-2.50) and serum sodium < or = 139 mEql/l (RR: 1.34; 1.10-2.10) increased the risk of stroke. In the CASTEL, stroke was the first cardiovascular cause of death. Some independent predictors usually unrelated to stroke mortality (namely pulse pressure, pre-diabetic IGT, UA and blood electrolytes disorders) have been identified. PMID- 12530770 TI - Body size, fat distribution, menarcheal age and blood pressure in 14-year-old girls. AB - BACKGROUND: The relationships between body size and fatness and blood pressure are generally acknowledged. The majority of the few studies that have examined the effect of fat distribution and maturation rate on blood pressure have used secondary sex characteristics as the measure of maturity. The aim of the present study is to examine the associations between blood pressure and relative weight, fat distribution, recalled menarcheal age and occurrence of menstruation (yes/no) in a sample comprising of 1149 14-year-old girls. METHODS: Systolic and diastolic blood pressure (DBP), height, weight and body circumferences were measured using standard protocols. Fatness was expressed as body mass index (BMI, kg/m2), whereas fat distribution was estimated by using waist-to-hip ratio. The girls' maturity status was assessed from exact recalled date of menarche. One-way analysis of covariance and multiple linear regression analyses were used to determine the strength of association among systolic blood pressure (SBP), DBP and BMI, menarcheal age and indices of fat distribution. RESULTS AND CONCLUSION: Height and BMI are significantly associated with SBP. Relative weight is the most important factor related to SBP independently of chronological age and maturity status. Height and age at menarche are significantly associated with DBP. Height of 14-year-old girls shows the same strength of association with SBP and DBP, whereas maturity status negatively correlates with DBP. Fat distribution shows no effect on the level of DBP in girls. PMID- 12530771 TI - Determinants of serum calcium in men and women. The Tromso Study. AB - The level of serum calcium appears to be associated with blood pressure and metabolic risk factors for cardiovascular disease. Determinants of serum calcium may therefore be of interest. In a health survey in Tromso in 1994-1995, 27,159 subjects were examined. The survey included measurements of serum calcium and questionnaires on diet and lifestyle factors. In males mean serum calcium declined from 2.41 mmol/l for those in their 20s to 2.34 mmol/l for those in their 80s. In females mean serum calcium was stable at a level of 2.35 mmol/l before the menopause, and thereafter reached a plateau of 2.39 mmol/l. In both sexes serum calcium showed a positive association with body mass index (BMI) and coffee consumption that persisted after correcting for other variables in a multiple regression model (p < 0.05). Physical activity had no significant association with serum calcium. In females alcohol consumption was negatively, and cigarette smoking positively associated with serum calcium (p < 0.01). No significant effect on the serum calcium levels was found for the intake of calcium or vitamin D, except for males with a calcium intake below 200 mg/day. Some of the observed effects, like the variation with age, may partly be explained by alterations in levels of serum albumin to which approximately 40% of circulating calcium is bound and which was not adjusted for in this study, whereas that is hardly the case for the association with BMI and coffee consumption. However, none of these factors could affect the serum calcium level more than 0.02 mmol/l, and the biological significance of the observed associations questionable. PMID- 12530772 TI - Characterisation of strains of enteroaggregative Escherichia coli isolated during the infectious intestinal disease study in England. AB - Strains of Escherichia coli, hybridising with a DNA probe for enteroaggregative E. coli (EAggEC), were isolated from patients with infectious intestinal disease (IID) or gastro-enteritis, and healthy controls during the study of IID in England. Of 3506 cases presenting with an IID, 160 (4.6%) had faecal EAggEC as compared with 46 (1.7%) of 2772 healthy controls, 53% of EAggEC isolated from each of the 'case' and the 'control' groups adhered in a 'stacked-brick' formation. Strains from cases and controls belonged to over 39 and 14 different serogroups respectively, and approximately half of the strains isolated did not react with antisera in the current somatic antigen serotyping scheme. Forty-nine cases with EAggEC (31%) had a known history of foreign travel. Over 50% of strains isolated from cases and controls were resistant to one or more of eight antimicrobials, and antimicrobial resistance was not statistically significantly more common among cases with a known history of foreign travel (p = 0.57). These data form part of the largest investigation carried out on these organisms in the UK to date and provide the most comprehensive analysis of strains of EAggEC isolated from the general population of England. PMID- 12530773 TI - Alcohol, smoking, coffee and risk of non-fatal acute myocardial infarction in Italy. AB - OBJECTIVES: To assess the relation between cigarette smoking, alcohol, coffee, decaffeinated coffee and tea consumption, and the risk of non-fatal acute myocardial infarction (AMI). DESIGN AND SETTING: Hospital-based case-control study conducted in 1995-1999 in Milan, Italy. PATIENTS: 507 cases with a first episode of non-fatal AMI, and 478 controls admitted to hospital for acute diseases. METHODS: Information was collected by interviewer-administered questionnaires. Odds ratios (OR) and 95% confidence intervals (CI) were calculated by multiple logistic regression. RESULTS: Compared to alcohol non drinkers the OR was 0.6 (95% CI: 0.4-0.9) in drinkers, and 0.5 in drinkers of > 3 drinks/day. The OR for > 1 drink/day of wine was 0.5, and those for beer, amari, grappa and spirits ranged between 0.4 and 0.6. Compared to never smokers, the OR was 2.2 (95% CI: 1.5-3.1) among current smokers, and 4.6 among current smokers of > or = 25 cigarettes/day. The risk was similar to that of never smokers > or = 5 years after cessation (OR: 1.1 after 5-9 years, 0.7 after > or = 10 years). The OR was 2.3 for low tar cigarettes and 2.0 for high tar ones. The OR for coffee intake (expresso and mocha) was around unity up to 3 cups/ day, but rose to 1.9 (95% CI: 1.1-3.3) for > or = 6 cups/ day. Moderate decaffeinated coffee and tea intake was not associated with AMI risk. Compared to non-smokers drinking < or = 3 cups of coffee/day, the OR was 1.6 among non-smokers drinking > 3 cups of coffee/ day and 3.3 (95% CI: 2.1-5.0) among current smokers drinking < or = 3 cups of coffee/day. Compared to alcohol drinkers with a coffee intake of < or = 3 cups/ day, alcohol non-drinkers with higher coffee intake had an OR of 2.2, and compared to non-smokers alcohol drinkers, the OR was 3.3 in current smokers alcohol non-drinkers. CONCLUSIONS: In this Italian population alcohol intake was inversely associated to AMI risk, while smoking and heavy (but not moderate) coffee drinking increased the risk. PMID- 12530774 TI - Trends in incidence, case-fatality and recurrence of myocardial infarction in the Danish MONICA population 1982-1991. AB - BACKGROUND: Growing awareness of risk factors for myocardial infarction (MI), continuous mass campaigns on lifestyle factors, increasing use of heart rehabilitation and improved treatment should imply decreases in the incidence, case-fatality and recurrence rates of MI. The purpose of the study was to assess trends in the incidence, case-fatality and recurrence rate of MI and to analyse whether any changes seen were due to a period or a cohort effect. METHODS: The Danish MONICA population comprises all men and women aged 25-74 years in the period 1982-1991 living in a suburban area of Copenhagen, Denmark. Fatal and non fatal attacks classified as definite MI and non-fatal attacks classified as possible MI were included. The incidence rate was analysed by Poisson regression, the case-fatality rate by logistic regression, and the rate of recurrence by Cox regression. Age-period-cohort analyses were carried out according to a method described by Clayton and Schifflers. RESULTS: During the 10-year period a significant decrease in the incidence rate of MI was seen for men and women and for the rate of recurrent MI. The decrease in incidence and recurrence could not unambiguously be ascribed to a period or cohort effect. The rate of case-fatality after a first MI was not changed significantly during the period, whereas men and women had different trends in case-fatality after recurrent MI. CONCLUSIONS: In accordance with results from other Western countries we found a decline in the incidence and recurrence rate of MI. Contrary, the lack of a decrease in the case fatality rate after a first MI was unexpected and difficult to explain. PMID- 12530775 TI - Letter to the Editor from Cicmanec. On the risk of mortality to primates exposed to anthrax spores. PMID- 12530776 TI - Risk communication is a key to dealing effectively with bioterrorism. PMID- 12530777 TI - Commentary on the conference on decision-making under conditions of uncertainty for rare diseases. PMID- 12530778 TI - Risk and systems theory. AB - The last few decades have seen increasingly widespread use of risk assessment and management techniques as aids in making complex decisions. However, despite the progress that has been made in risk science, there still remain numerous examples of risk-based decisions and conclusions that have caused great controversy. In particular, there is a great deal of debate surrounding risk assessment: the role of values and ethics and other extra-scientific factors, the efficacy of quantitative versus qualitative analysis, and the role of uncertainty and incomplete information. Many of the epistemological and methodological issues confronting risk assessment have been explored in general systems theory, where techniques exist to manage such issues. However, the use of systems theory and systems analysis tools is still not widespread in risk management. This article builds on the Alachlor risk assessment case study of Brunk, Haworth, and Lee to present a systems-based view of the risk assessment process. The details of the case study are reviewed and the authors' original conclusions regarding the effects of extra-scientific factors on risk assessment are discussed. Concepts from systems theory are introduced to provide a mechanism with which to illustrate these extra-scientific effects The role of a systems study within a risk assessment is explained, resulting in an improved view of the problem formulation process The consequences regarding the definition of risk and its role in decision making are then explored. PMID- 12530779 TI - Influence of safety measures on the risks of transporting dangerous goods through road tunnels. AB - Quantitative risk assessment (QRA) models are used to estimate the risks of transporting dangerous goods and to assess the merits of introducing alternative risk reduction measures for different transportation scenarios and assumptions. A comprehensive QRA model recently was developed in Europe for application to road tunnels. This model can assess the merits of a limited number of "native safety measures." In this article, we introduce a procedure for extending its scope to include the treatment of a number of important "nonnative safety measures" of interest to tunnel operators and decisionmakers. Nonnative safety measures were not included in the original model specification. The suggested procedure makes use of expert judgment and Monte Carlo simulation methods to model uncertainty in the revised risk estimates. The results of a case study application are presented that involve the risks of transporting a given volume of flammable liquid through a 10-km road tunnel. PMID- 12530780 TI - A new approach to risk evaluation and management: risk-based, precaution-based, and discourse-based strategies. AB - Our concept of nine risk evaluation criteria, six risk classes, a decision tree, and three management categories was developed to improve the effectiveness, efficiency, and political feasibility of risk management procedures. The main task of risk evaluation and management is to develop adequate tools for dealing with the problems of complexity, uncertainty. and ambiguity. Based on the characteristics of different risk types and these three major problems, we distinguished three types of management--risk-based, precaution-based, and discourse-based strategies. The risk-based strategy--is the common solution to risk problems. Once the probabilities and their corresponding damage potentials are calculated, risk managers are required to set priorities according to the severity of the risk, which may be operationalized as a linear combination of damage and probability or as a weighted combination thereof. Within our new risk classification, the two central components have been augmented with other physical and social criteria that still demand risk-based strategies as long as uncertainty is low and ambiguity absent. Risk-based strategies are best solutions to problems of complexity and some components of uncertainty, for example, variation among individuals. If the two most important risk criteria, probability of occurrence and extent of damage, are relatively well known and little uncertainty is left, the traditional risk-based approach seems reasonable. If uncertainty plays a large role, in particular, indeterminacy or lack of knowledge, the risk-based approach becomes counterproductive. Judging the relative severity of risks on the basis of uncertain parameters does not make much sense. Under these circumstances, management strategies belonging to the precautionary management style are required. The precautionary approach has been the basis for much of the European environmental and health protection legislation and regulation. Our own approach to risk management has been guided by the proposition that any conceptualization of the precautionary principle should be (1) in line with established methods of scientific risk assessments, (2) consistent and discriminatory (avoiding arbitrary results) when it comes to prioritization, and (3) at the same time, specific with respect to precautionary measures, such as ALARA or BACT, or the strategy of containing risks in time and space. This suggestion does, however, entail a major problem: looking only to the uncertainties does not provide risk managers with a clue about where to set priorities for risk reduction. Risks vary in their degree of remaining uncertainties. How can one judge the severity of a situation when the potential damage and its probability are unknown or contested? In this dilemma, we advise risk managers to use additional criteria of hazardousness, such as "ubiquity versibility," and "pervasiveness over time," as proxies for judging severity. Our approach also distinguishes clearly between uncertainty and ambiguity. Uncertainty refers to a situation of being unclear about factual statements; ambiguity to a situation of contested views about the desirability or severity of a given hazard. Uncertainty can be resolved in principle by more cognitive advances (with the exception of indeterminacy). ambiguity only by discourse. Discursive procedures include legal deliberations as well as novel participatory approaches. In addition, discursive methods of planning and conflict resolution can be used. If ambiguities are associated with a risk problem, it is not enough to demonstrate that risk regulators are open to public concerns and address the issues that many people wish them to take care ot The process of risk evaluation itself needs to be open to public input and new forms of deliberation. We have recommended a tested set of deliberative processes that are, at least in principle, capable of resolving ambiguities in risk debates (for a review, see Renn, Webler, & Wiedemaun. 1995). Deliberative processes are needed, however, for ail three types of management. Risk-based management relies on epistemiological, uncertainty-based management on reflective, and discourse-based management on participatory discourse forms. These three types of discourse could be labeled as an analytic-deliberative procedure for risk evaluation and management. We see the advantage of a deliberative style of regulation and management in a dynamic balance between procedure and outcome. Procedure should not have priority over the outcome; outcome should not have priority over the procedure. An intelligent combination of both can elaborate the required prerequisites of democratic deliberation and its substantial outcomes to enhance the legitimacy of political decisions (Guttman & Thompson, 1996; Bohman, 1997. 1998). PMID- 12530781 TI - Health risk assessment for planned waste incinerators: getting the right science and the science right. AB - Health risk assessment is widely advocated in the United Kingdom as the most comprehensive means of assessing the health risks posed by the emissions of a planned waste incinerator. Its main advantage over other methods of assessment, such as air quality impact assessment, is its ability to address explicitly the direct (inhalation) and indirect (ingestion and dermal contact) health risks posed by different chemicals, including those that are not thought to have a threshold below which no adverse effect will take place. This article examines the level and quality of the emissions assessments included in 61 waste incinerator environmental statements (ESs); in particular, it focuses on the quality of the exposure assessment and risk characterization stages of the health risk assessment process. The article concludes that the ES has not always provided interested stakeholders with the best available information upon which to determine the tolerability of the health risks posed by waste incinerator emissions Some recommendations are made as to how this problem might be addressed in future environmental impact assessment (EIA) processes. PMID- 12530782 TI - An empirical test of the relative validity of expert and lay judgments of risk. AB - This article investigates how accurately experts (underwriters) and lay persons (university students) judge the risks posed by life-threatening events Only one prior study (Slovic, Fischhoff, & Lichtenstein, 1985) has previously investigated the veracity of expert versus lay judgments of the magnitude of risk. In that study, a heterogeneous grouping of 15 experts was found to judge, using marginal estimations, a variety of risks as closer to the true annual frequencies of death than convenience samples of the lay population. In this study, we use a larger, homogenous sample of experts performing an ecologically valid task. We also ask our respondents to assess frequencies and relative frequencies directly, rather than ask for a "risk" estimate--a response mode subject to possible qualitative attributions-as was done in the Slovic et al. study. Although we find that the experts outperformed lay persons on a number of measures, the differences are small, and both groups showed similar global biases in terms of: (1) overestimating the likelihood of dying from a condition (marginal probability) and of dying from a condition given that it happens to you (conditional probability), and (2) underestimating the ratios of marginal and conditional likelihoods between pairs of potentially lethal events. In spite of these scaling problems, both groups showed quite good performance in ordering the lethal events in terms of marginal and conditional likelihoods. We discuss the nature of expertise using a framework developed by Bolger and Wright (1994), and consider whether the commonsense assumption of the superiority of expert risk assessors in making magnitude judgments of risk is, in fact, sensible. PMID- 12530783 TI - Stakeholder attitudes toward the risks and benefits of agricultural biotechnology in developing countries: a comparison between Mexico and the Philippines. AB - Public perceptions and political debates regarding the risks and benefits of agricultural biotechnology are well documented in industrialized countries. Yet, hardly any surveys have been conducted in developing countries. The following study aims to contribute to a better understanding of stakeholder attitudes toward agricultural biotechnology in developing countries. For this purpose, stakeholder representatives in the public debates in Mexico and the Philippines were asked to complete two nearly identical questionnaires on the risks and benefits of agricultural biotechnology. A comparison of stakeholder attitudes in the Philippines and Mexico is interesting because it shows how the different political systems, cultural and ecological backgrounds, and the regional context of food and agriculture influence perceptions. In general, the results of the surveys indicate that the participants in both countries consider genetic engineering to be an important tool to address agricultural, nutritional, and environmental problems, and they do not regard transgenic foods as risky for consumers. However, they are concerned about the potential impact of such transgenic crops on their countries' rich biological diversity and do not believe that national biosafety guidelines will be implemented properly. Although the surveys show that stakeholder attitudes in the Philippines and Mexico are quite similar, they also highlight significant differences in perception often related to cultural and political aspects. PMID- 12530784 TI - Measuring perceptions of synergistic circulatory disease risk due to smoking and the oral contraceptive pill. AB - There is evidence that the oral contraceptive pill (OCP) and smoking contribute independently to risk of circulatory disease. There is mixed evidence that the combined risk may be greater than the sum of these factors operating in isolation. Little is known about how the general population views the risks from OCP use, singly and in combination with smoking. Previous attempts at assessing whether the public views risks as operating synergistically have generally found evidence for subadditive models, where the combined risk is less than the sum of factors operating in isolation. However, concerns have been expressed over the validity of the measures of risk perception used. Therefore, this study used three distinct methods of measurement to assess the extent to which 241 undergraduate students perceive the risks of smoking and the OCP separately and combined, for circulatory disease. For all three methods, respondents read each of four vignettes describing information about a woman's risk factors (with high and low levels of both OCP and smoking), and then estimated risk of circulatory disease using one of the three risk measures. The three measures produced similar ratings. Consistent with the epidemiological evidence, information about smoking had more impact on estimates of overall risk than did information about the OCP For all three measures, responses were consistent with an additive model of risk from smoking and the OCP. This convergence of results from different methods suggests that all three methods of measurement employed, which all had a large number of response options, may be valid. PMID- 12530785 TI - A risk analysis for airborne pathogens with low infectious doses: application to respirator selection against Coccidioides immitis spores. AB - Probability models incorporating a deterministic versus stochastic infectious dose are described for estimating infection risk due to airborne pathogens that infect at low doses. Such pathogens can be occupational hazards or candidate agents for bioterrorism. Inputs include parameters for the infectious dose model, distribution parameters for ambient pathogen concentrations, the breathing rate, the duration of an exposure period, the anticipated number of exposure periods, and, if a respirator device is used, distribution parameters for respirator penetration values. Application of the models is illustrated with a hypothetical scenario involving exposure to Coccidioides immitis, a fungus present in soil in areas of the southwestern United States Inhaling C. immitis spores causes a respiratory tract infection and is a recognized occupational hazard in jobs involving soil dust exposure in endemic areas An uncertainty analysis is applied to risk estimation in the context of selecting respiratory protection with a desired degree of efficacy. PMID- 12530786 TI - Joint models for toxicology studies with dose-dependent number of implantations. AB - Many chemicals interfere with the natural reproductive processes in mammals. The chemicals may prevent the fertilization of an egg or keep a zygote from implanting in the uterine wall. For this reason, toxicology studies with pre implantation exposure often exhibit a dose-related trend in the number of observed implantations per litter. Standard methods for analyzing developmental toxicology studies are conditioned on the number of implantations in the litter and therefore cannot estimate this effect of the chemical on the reproductive process. This article presents a joint modeling approach to estimating risk in toxicology studies with pre-implantation exposure. In the joint modeling approach, both the number of implanted fetuses and the outcome of each implanted fetus is modeled. Using this approach we show how to estimate the overall risk of a chemical that incorporates the risk of lost implantation due to pre implantation exposure. Our approach has several distinct advantages over previous methods: (1) it is based on fitting a model for the observed data and, therefore, diagnostics of model fit and selection apply; (2) all assumptions are explicitly stated; and (3) it can be fit using standard software packages We illustrate our approach by analyzing a dominant lethal assay data set (Luning et al., 1966, Mutation Research, 3, 444-451) and compare ourresults with those of Rai and Van Ryzin (1985, Biometrics, 41,1-9) and Dunson (1998, Biometrics, 54, 558-569). In a simulation study, our approach has smaller bias and variance than the multiple imputation procedure of Dunson. PMID- 12530787 TI - Uncertainty in measurements of dermal absorption of pesticides. AB - Dermal absorption experiments form an important component in the assessment of risk from exposure to pesticides and other substances. Much dermal absorption data is gathered in rat experiments carried out using a certain standard protocol. Uncertainties in these data arise from many sources and can be quite large. For example, measurements of the systemic absorption of hexaconazole differed by more than an order of magnitude within a single experiment. Two diniconazole studies produced quite different results, due to minor differences in protocol and in chemical formulation. Limits of detection can also prevent accurate measurement when the amounts absorbed are small. These examples illustrate the need for measuring and reporting uncertainties in estimates that are based on these data. The most direct way to estimate uncertainty is to compute the sample standard deviations of replicate measurements. By pooling these estimates across dose and duration groups for which they are similar, the number of degrees of freedom is increased, and more precise confidence intervals can be obtained. In particular, the ratio of upper to lower 95% confidence limits was reduced by as much as ten-fold for hexaconazole, seven-fold for uniconazole, and nearly four-fold for propiconazole. PMID- 12530788 TI - Exposure misclassification and threshold concentrations in time series analyses of air pollution health effects. AB - Linear, no-threshold relationships are typically reported for time series studies of air pollution and mortality. Since regulatory standards and economic valuations typically assume some threshold level, we evaluated the fundamental question of the impact of exposure misclassification on the persistence of underlying personal-level thresholds when personal data are aggregated to the population level in the assessment of exposure-response relationships. As an example, we measured personal exposures to two particle metrics, PM2.5 and sulfate (SO4(2-)), for a sample of lung disease patients and compared these with exposures estimated from ambient measurements Previous work has shown that ambient:personal correlations for PM2.5 are much lower than for SO4(2-), suggesting that ambient PM2.5 measurements misclassify exposures to PM2.5. We then developed a method by which the measured:estimated exposure relationships for these patients were used to simulate personal exposures for a larger population and then to estimate individual-level mortality risks under different threshold assumptions. These individual risks were combined to obtain the population risk of death, thereby exhibiting the prominence (and the value) of the threshold in the relationship between risk and estimated exposure. Our results indicated that for poorly classified exposures (PM2.5 in this example) population-level thresholds were apparent at lower ambient concentrations than specified common personal thresholds, while for well-classified exposures (e.g., SO4(2-)), the apparent thresholds were similar to these underlying personal thresholds. These results demonstrate that surrogate metrics that are not highly correlated with personal exposures obscure the presence of thresholds in epidemiological studies of larger populations, while exposure indicators that are highly correlated with personal exposures can accurately reflect underlying personal thresholds. PMID- 12530789 TI - Optimal designs for estimating the effective dose in developmental toxicity experiments. AB - Recent advances in risk assessment have led to the development of joint dose response models to describe prenatal death and fetal malformation rates in developmental toxicity experiments. These models can be used to estimate the effective dose corresponding to a 5% excess risk for both these toxicological endpoints, as well as for overall toxicity. In this article, we develop optimal experimental designs for the estimation of the effective dose for developmental toxicity using joint Weibull dose-response models for prenatal death and fetal malformation. Based on an extended series of developmental studies, near-optimal designs for prenatal death, malformation, and overall toxicity were found to involve three dose groups: an unexposed control group, a high dose equal to the maximum tolerated dose, and a low dose above or comparable to the effective dose. The effect on the optimal designs of changing the number of implants and the degree of intra-litter correlation is also investigated. Although the optimal design has only three dose groups in most cases, practical considerations involving model lack of fit and estimation of the shape of the dose-response curve suggest that, in practice, suboptimal designs with more than three doses will often be preferred. PMID- 12530790 TI - Comparing indigenous health status across regions: a numerical example of uncertainty. AB - OBJECTIVE: To illustrate how regional variation in data quality could explain some or all of the apparent regional differences in the health status of Indigenous Australians. METHODS: A series of simple hypothetical numerical examples is provided, with varying assumptions regarding the accuracy of identification of Indigenous deaths. RESULTS: The apparent difference in Indigenous mortality in remote compared with urban areas is of a magnitude that could be explained by relatively modest regional differences in data quality. CONCLUSION AND IMPLICATIONS: Determinations of relative health status within the Indigenous population must take into account the impact of variability in data quality. PMID- 12530791 TI - Performance of diagnosis-based risk adjustment measures in a population of sick Australians. AB - OBJECTIVE: Australia is beginning to explore 'managed competition' as an organising framework for the health care system. This requires setting fair capitation rates, i.e. rates that adjust for the risk profile of covered lives. This paper tests two US-developed risk adjustment approaches using Australian data. METHODS: Data from the 'co-ordinated care' dataset (which incorporates all service costs of 16,538 participants in a large health service research project conducted in 1996-99) were grouped into homogenous risk categories using risk adjustment 'grouper software'. The grouper products yielded three sets of homogenous categories: Diagnostic Groups and Diagnostic cost Groups. A two-stage analysis of predictive power was used: probability of any service use in the concurrent year, next year and the year after (logistic regression) and, for service users, a regression of logged cost of service use. The independent variables were diagnosis gender, a SES variable and the RESULTS: Age, gender and diagnosis-based risk adjustment measures explain around 40-45% of variation in costs of service use in the current year for untrimmed data (compared with around 15% for age and gender alone). Prediction of subsequent use is much poorer (around 20%). Using more information to assign people to risk categories generally improves prediction. CONCLUSIONS: Predictive power of diagnosis-base risk adjusters on this Australian dataset is similar to that found in IMPLICATIONS: Low predictive power carries policy risks of cream skimming rather than managing population health and care. Competitive funding models with risk adjustment on prior year experience could reduce system efficiency if implemented with current risk adjustment technology. PMID- 12530792 TI - A brief integer scorer for the SF-12: validity of the brief scorer in Australian community and clinic settings. AB - OBJECTIVE: The SF-36 and the shorter SF-12 are widely used in clinical research and increasingly so in practice. Scoring is complex and the validity of a brief scorer for the SF-12 in a community and a clinic sample was examined. METHOD: The sample comprised respondents with complete data on the SF-12 from the community sample in the 1997 Australian National Survey of Mental Health and Well-being (n = 10,641) and from a clinical sample from the Clinical Research Unit for Anxiety and Depression (n = 1,725). Each SF-12 was scored by the standard method (using weights to five decimal places) or by the brief method (using weights that are integers only). RESULTS: Scores generated by the standard scorer correlated almost perfectly with the brief scorer in both the community and clinic samples. Means and standard deviations were similar and no individual scores deviated by more than 2.89 in the community sample or by 3.06 in the clinical sample. IMPLICATIONS: The brief, rounded integer scorer for the SF-12 is an appropriate substitute for the standard scorer when hand scoring is an advantage. PMID- 12530793 TI - The experience of using random digit dialling methods in a population-based chronic pain study. AB - OBJECTIVES: To describe response rates and call characteristics using random digit dialling (RDD) methods in a population-based prevalence study of chronic pain; to compare respondent characteristics according to telephone number listing status. METHODS: A RDD telephone study of chronic pain was conducted using computer-assisted telephone interview (CATI) methods in the Northern Sydney Area in 1998. RESULTS: Unlisted number respondents (18.5%) were younger, had a higher proportion of males, and were less likely to live in households with three or more eligible adults. There was no difference in chronic pain status between the two groups. The overall response rate was 73.4% (unlisted number group 66.3%; listed number group 75.0%). Answering machine messages boosted response rates (79.7%). 10.9% of unlisted numbers used in the study resulted in completed interviews, compared with 31.3% of the listed numbers used. CONCLUSIONS: The relatively high response rate obtained, differences in characteristics associated with listing status and reduction in sampling bias provided by using RDD methods have to be balanced against the differential response rates between listed and unlisted number groups and higher costs (including opportunity costs). IMPLICTIONS: Published data on the experience of using RDD methods can assist public health researchers in deciding whether to use these methods in telephone surveys. PMID- 12530794 TI - Australia's food regulations--challenges for public health advocacy. AB - OBJECTIVE: To illustrate current challenges to public health advocacy through examining the case of the development of food regulations in Australia. METHOD: To present a case study of the recent changes to the structures for the development of food regulations in Australia and aspects of the new Food Standards Code that have direct relevance to public health. RESULTS: Clear links between the policy setting structure and some types of public health advocacy strategies are made. CONCLUSIONS AND RECOMMENDATIONS: The challenge for public health advocacy is to ensure that professional associations as well as individual health professionals are cognisant of the environment within which public policy is developed, as well as being able to professionally comment on the content of public policy, for positive impacts on public health priorities to be achieved. PMID- 12530795 TI - Public health aspects of genetic screening for hereditary haemochromatosis in Australia. AB - Hereditary haemochromatosis (HH) is an inherited disorder of iron absorption. It meets several of the key public health principles for population-based screening and is considered to be a test-case for public health genetics. However, there has been relatively little debate in the public health or wider community regarding the merits of population-based genetic screening for HH. Genetic susceptibility to HH occurs in about 1:200 people and although mortality is low (age-standardised rate 2.75/million), there are potentially serious clinical manifestations of iron overload. Regular venesection is a simple and effective treatment for early stage iron overload. DNA-based testing is available and iron overload may be identified using serum transferrin saturation and ferritin tests. However, there are important gaps in knowledge relevant to screening for HH. The limited data on penetrance of HFE genotypes, and thus the uncertain probability that genetically susceptible individuals will develop clinically significant disease, is a major impediment to population-based genetic screening. Clinical evidence supports treating early-stage disease but no randomised controlled trials of the effectiveness of screening in reducing the burden of disease have been conducted. In addition, the natural history of early stages of HH and factors that may modify progression are unclear. Two intemational consensus panels on HH concluded that there is insufficient evidence for population-based screening at present. We present recommendations to advance the debate on screening for HH in Australia. PMID- 12530796 TI - Were NZ's structural changes to the welfare state in the early 1990s associated with a measurable increase in oral health inequalities among children? AB - OBJECTIVE: To test the hypothesis that the 1990-91 social and economic policy changes in New Zealand were associated with a subsequent increase in socioeconomic and ethnic inequalities in the dental caries experience of five year-old children. METHOD: Dental caries data from the School Dental Service treating the greater Wellington area were analysed for the period 1995-2000. Multivariate models were developed for deciduous caries prevalence (logistic regression) and severity (negative binomial regression). RESULTS: In the years 1995, 1996, 1997, 1998, 1999 and 2000, complete data were available for 2,627, 3,335, 4,404, 4,155, 3,154 and 2,804 children, respectively. Ethnic and socio economic differences in caries prevalence and severity were substantial and persistent during the observation period. Where caries severity was concerned, there was a significant interaction between time and Maori ethnicity, indicating that (on average) the oral health of Maori children deteriorated in comparison to their European counterparts. CONCLUSIONS: The early-1990s social and economic policy changes were associated with an apparent widening of ethnic inequalities in caries severity among five-year-old children. IMPLICATIONS: Economic rationalism appears to have oral health disadvantages for non-European children. Before implementation of proposed major social and economic policy changes, policymakers should consider their health implications. PMID- 12530797 TI - Inter-agency collaboration: the North East Health Promotion Centre. AB - OBJECTIVE: The management of interagency collaborations needs to be approached in particular, but not well defined, ways. In this paper the management and ways of working of a successful co-ordinating unit are discussed and located in the context of inter-agency partnerships. METHODS: The text is based on qualitative archival research and a critical review of practice. CONCLUSIONS: The evidence indicates that the agencies participating in the centre valued its work in facilitating inter-agency collaboration. The structural arrangements of the centre, and its modus operandi, manifest the characteristics of an 'obligational co-ordinating unit' in which success is based on establishing and maintaining appropriate interactions between partner agencies. The ways the centre operationalised the key principles of independence, member support, power and trust relations are discussed. Finally, the importance of 'environmental fit' and the impact of a changing policy environment on the centre are discussed. IMPLICATIONS: In health promotion, and public health more broadly, efforts to co ordinate the work of agencies and their members are common. However, the precise structures and work practices that are effective rarely have a foundation in evidence or theory. This paper will contribute to our knowledge of how the work of co-ordination can be effectively undertaken in the context of agencies voluntarily entering negotiated relationships to pursue joint goals in public health. PMID- 12530798 TI - Environmental tobacco smoke in homes, motor vehicles and licensed premises: community attitudes and practices. AB - OBJECTIVE: To assess community knowledge, attitudes and practices in relation to environmental tobacco smoke (ETS) especially in homes, private motor vehicles and licensed premises, and to document levels of support for further government legislation. METHODS: 656 persons aged 18 years and over, a sub-sample in a computer-assisted telephone survey of 2,087 randomly selected respondents across NSW, answered 12 ETS-specific questions. The whole sample was asked demographic, smoking status and household membership questions. The overall response rate was 61.4% (consent rate 75.4%). RESULTS: Overall, 32.8% (95% CI 27.8-37.8) of children aged 0-4 years in the households surveyed were reported to live with at least one smoker. Agreement about ETS hazards was lowest in relation to child ear problems (31.2%) and sudden infant death syndrome (50.6%). Complete bans on smoking were reported in 69.9% of homes and 77.1% of private motor vehicles. The percentages favouring total bans in homes (p < 0.001) and private motor vehicles (p < 0.001) were significantly lower among smokers than non-smokers. Overall, 55.8% supported legislation to ban smoking in private vehicles carrying children. The majority support restrictions on smoking in non-eating areas of licensed clubs (88.7%) and hotels (84.8%). Overall, the data indicate a ban on smoking in licensed premises is likely to increase business. CONCLUSION: Smoking is banned in most NSW homes and private motor vehicles. Acceptance of ETS risks is high but there are important knowledge gaps. Considerable support exists for further government regulation of ETS. Media campaigns are likely to reinforce a public environment already receptive of the need for more government ETS restrictions. PMID- 12530799 TI - Hospital system costs of artificial infant feeding: estimates for the Australian Capital Territory. AB - OBJECTIVE: To estimate the attributable ACT hospital system costs of treating selected infant and childhood illnesses having known associations with early weaning from human milk. METHOD: We identified relative risks of infant and childhood morbidity associated with exposure to artificial feeding in the early months of life vs. breastfeeding from cohort studies cited by the American Academy of Pediatrics in 1997 as establishing the protective effect of breastfeeding. Data for ACT breastfeeding prevalence is assessed from a 1997 prospective population-based cohort study of 1,295 women. ACT Hospital Morbidity Data and DRG treatment costs were used to estimate the attributable fraction of costs of hospitalisation for gastrointestinal illness, respiratory illness and otitis media, eczema, and necrotising enterocolitis. RESULTS: Although initiation rates were high (92%), less than one in 10 ACT infants are exclusively breastfed for the recommended six months, mainly due to supplementation or weaning on to formula within the first three months and the early introduction of solids by breastfeeding mothers. This study suggests the attributable hospitalisation costs of early weaning in the ACT are about $1-2 million a year for the five illnesses. CONCLUSIONS AND IMPLICATIONS: Early weaning from breastmilk is associated with significant hospital costs for treatment of gastrointestinal illness, respiratory illness and otitis media, eczema, and necrotising enterocolitis. These costs are minimum estimates of the cost of early weaning as they exclude numerous other chronic or common illnesses and out-of-hospital health care costs. Higher rates of exclusive breastfeeding would reduce these costs. Interventions to protect and support breastfeeding are likely to be cost-effective for the public health system. PMID- 12530800 TI - The Sydney Medically Supervised Injecting Centre: a controversial public health measure. AB - BACKGROUND: Injecting drug use remains a major public health concern, particularly because of opiate overdose and transmission of blood-borne viruses. Sydney's Medically Supervised Injecting Centre (MSIC) opened on a trial basis in May 2001 in an effort to reduce the harms of drug use. In this report, we provide a brief overview of the reported public health impact of supervising injecting facilities (SIFs) and review the history and early process evaluations of the Sydney Centre. METHODS: Medline, Internet searches and perusal of bibliographies of articles were used to identify key English language publications on SIFs. These were supplemented by interview with the Medical Director of Sydney MSIC, Dr Ingrid van Beek. DISCUSSION AND CONCLUSIONS: It is difficult to be certain of the public health impact of SIFs but evidence from overseas and Sydney's early process evaluations provide promise that they may make a positive contribution to health. PMID- 12530801 TI - Risk and protective factors for depressive symptomatology among a community sample of adolescents and young adults. AB - OBJECTIVES: The study explores the risk and protective factors for current depressive symptomatology in a large community sample of 15-to-24-year-olds. METHODS: The study was designed as a cross-sectional household survey, which used telephone recruitment followed by an anonymous self-report postal questionnaire. The final sample included 3,082 adolescents and young adults from Queensland, Australia. RESULTS: The vast majority of measured risk and protective factors were associated with current depressive symptomatology. Key risk factors included high levels of neuroticism, perceived problems with parents, sexual abuse, relationship break-ups, educational failure and sexual identity conflict. A different profile of protective factors was evident for each of these high-risk groups. Of particular note was the importance of well-developed intrapersonal skills as protective for both males and females. The significance of social connectedness as a protective factor for the males overall and across a range of high-risk groups was a central finding. CONCLUSIONS AND IMPLICATIONS: The implications of these findings in relation to a range of mental health promotion and mental illness prevention and early intervention initiatives are discussed. Supported initiatives include parenting programs that consider the realities of modem families, increasing community awareness of the impact on young people of the breakdown of their intimate relationships, initiatives in educational settings and workplaces to increase tolerance of gay/lesbian and bisexual lifestyles and the enhancement of social connectedness. PMID- 12530802 TI - Menopausal transitions, symptoms and country of birth: the Australian Longitudinal Study on Women's Health. AB - OBJECTIVE: To assess differences among the menopausal transitions and symptoms experienced by women participating in the Australian Longitudinal Study on Women's Health, according to their countries of birth. METHODS: Data from 8,466 women aged 45-50 in 1996, who responded to surveys in 1996 and 1998 and had not had a hysterectomy, were analysed. Women were categorised by country of birth and cross-sectionally by menopausal status at Survey 1 and 2, as well as longitudinally by transition through menopause between Surveys 1 and 2. Four endocrine-related and 10 general symptoms were assessed. RESULTS: Women bom in Asia were twice as likely as Australian-bom women to be post-menopausal at Survey 1, twice as likely to become post-menopausal between surveys, less likely to remain peri-menopausal, and less likely to report hot flushes and night sweats. Odds ratios for each symptom at Survey 2 were near unity for all country of birth groups compared with Australian bom women, with or without adjustment for symptoms at Survey 1, menopausal transition category, behaviour, lifestyle and demographics. CONCLUSIONS: Asian-born women entered menopause earlier and passed through it more quickly, but once this was taken into account all women showed the same prevalences of symptoms. There may be differences between ethnic groups that influence the timing of menopause, but the subjective experience appears similar. IMPLICATIONS: The timing of menopause may be affected by biological or dietary differences. Asian-bom women's lower reported prevalence of symptoms may be explained by a more rapid peri-menopausal transition. With increasing numbers of Asian-Australian women reaching menopause, an understanding of country-of birth differences has implications for public health. PMID- 12530803 TI - Tobacco use of pregnant women and their male partners who attend antenatal classes: what happened to routine quit smoking advice in pregnancy? PMID- 12530804 TI - Trojan horses: how the tobacco industry infiltrates the smokefree debate in Australia. PMID- 12530806 TI - Chips and more chips. PMID- 12530807 TI - Unified chromatography with CO2-based binary mobile phases. PMID- 12530805 TI - Environmental health and social factors associated with motor neurone disease. PMID- 12530808 TI - Chiral analysis by MS. PMID- 12530810 TI - The endearing FTIR spectrophotometer. PMID- 12530809 TI - The quest for new prion tests. PMID- 12530811 TI - Continuous segmented-flow polymerase chain reaction for high-throughput miniaturized DNA amplification. AB - A continuous segmented-flow method for sequential DNA amplification is described in order to provide a basis for high-throughput genetic analysis. The approach allows an immediate distinction between amplified and nonamplified products. A mixture of sample and reagents are loaded in the form of small segments one after another in a 15-m-long narrow-bore Teflon tube, coiled such as to be repeatedly exposed to three different temperature zones. After having passed the heated zones, the samples are mixed with an intercalating dye by flow injection and sequentially detected on-line by laser-induced fluorescence. The aqueous samples travel as separate segments in a continuous flow of an immiscible, organic liquid. Perfluorodecalin was shown to be particularly suitable due to its hydrophobicity and inert properties. To reduce carryover between samples, an intermediate water plug between two consecutive samples was required. Selected regions from human genomic DNA were successfully amplified in 300-nL volumes after 30 passes through the heated zones. The total reaction time was approximately 45 min, and the detection interval between individual samples was 1 min. Automation and the possibility to further reduce sample volumes, as well as to employ many reaction columns simultaneously, should provide a platform for an extremely high throughput. PMID- 12530812 TI - Distribution of zeptomole-abundant doxorubicin metabolites in subcellular fractions by capillary electrophoresis with laser-induced fluorescence detection. AB - Doxorubicin (DOX) treatment of NS-1 mouse hybridoma cells results in the formation of zeptomole amounts of metabolites per cell that are difficult to determine by confocal microscopy or HPLC. The native fluorescence of DOX and its metabolites together with laser-induced fluorescence detection (HF) has previously been used to detect a maximum of four components. In this study, we use capillary electrophoresis with postcolumn LIF (CE-LIF) to separate and detect 12 components attributed to DOX metabolism, resulting from treatment of NS-1 cells with 25 microM DOX for 8 h. The so-called metabolites 8 and 10 have been identified as doxorubicinone (DOXone) and 7-deoxydoxorubicinone (7-deoxyDOXone), respectively, by comigration with the corresponding synthetic standard. Due to comigration of DOX with doxorubicinol (DOXone), the presence of DOXone had to be determined separately by matrix-assisted laser desorption/ionization time-of flight mass spectrometry. The rest of the metabolites remain unidentified and are referred to by their number assignment. In comparison with the whole cell lysate, fractionation by differential centrifugation results in a better separation resolution of metabolites due to reduced amounts of metabolites in each fraction. This approach was chosen to compare the distribution of 13 metabolites in three subcellular fractions that form a pellet at < 1,400 g, 1,400-14,000 g, and > 14, 000 g and that generically are enriched in nuclei, organelles (mitochondria and lysosomes), and cytosolic components, respectively. The most abundant metabolite, DOXone, was estimated to be 90 +/- 15, 18 +/- 2, and 60 +/- 12 amol/cell (n = 5) in the nuclear-enriched, organelle-enriched, and cytosole-enriched fractions, respectively. In contrast, the total amount of other metabolites in a given fraction varied from 0 to 1,300 zmol. 7-DeoxyDOXone is the only metabolite that was present at similar levels in the three fractions. Other salient observations are metabolites 3, 7, and 11 are not detectable in the nuclear-enriched, organelle-enriched, and cytosole-enriched fractions, respectively; metabolite 9 and DOXone are more abundant in the nuclear-enriched fraction than in the other two fractions. The observations presented here suggest that subcellular fractionation followed by CE-LIF could be a powerful diagnostic for monitoring drug distribution, which is highly relevant to DOX cytoxicity studies. PMID- 12530813 TI - Colloidal gold-modified optical fiber for chemical and biochemical sensing. AB - A novel class of fiber-optic evanescent-wave sensor was constructed on the basis of modification of the unclad portion of an optical fiber with self-assembled gold colloids. The optical properties and, hence, the attenuated total reflection spectrum of self-assembled gold colloids on the optical fiber changes with different refractive index of the environment near the colloidal gold surface. With sucrose solutions of increasing refractive index, the sensor response decreases linearly. The colloidal gold surface was also functionalized with glycine, succinic acid, or biotin to enhance the selectivity of the sensor. Results show that the sensor response decreases linearly with increasing concentration of each analyte. When the colloidal gold surface was functionalized with biotin, the detection limit of the sensor for streptavidin was 9.8 x 10(-11) M. Using this approach, we demonstrate proof-of-concept of a class of refractive index sensor that is sensitive to the refractive index of the environment near the colloidal gold surface and, hence, is suitable for label-free detection of molecular or biomolecular binding at the surface of gold colloids. PMID- 12530815 TI - The SLIM spectrometer. AB - A new spectrometer, here denoted the SLIM (simple, low-power, inexpensive, microcontroller-based) spectrometer, was developed that exploits the small size and low cost of solid-state electronic devices. In this device, light-emitting diodes (LED), single-chip integrated circuit photodetectors, embedded microcontrollers, and batteries replace traditional optoelectronic components, computers, and power supplies. This approach results in complete customizable spectrometers that are considerably less expensive and smaller than traditional instrumentation. The performance of the SLIM spectrometer, configured with a flow cell, was evaluated and compared to that of a commercial spectrophotometer. Thionine was the analyte, and the detection limit was approximately 0.2 microM with a 1.5-mm-path length flow cell. Nonlinearity due to the broad emission profile of the LED light sources is discussed. PMID- 12530814 TI - A microfluidic bioreactor based on hydrogel-entrapped E. coli: cell viability, lysis, and intracellular enzyme reactions. AB - Viable E. coli cells were entrapped in hydrogel micropatches photopolymerized within microfluidic systems. The microfluidic channels and the micropatches have sizes on the order of 100-500 microm. Small molecules, such as dyes and surfactants, present in the solution surrounding the hydrogel, are able to diffuse into the gel and encounter the cells, but the cells are sufficiently large to be retained. For example, sodium dodecyl sulfate is a lysis agent that is able to penetrate the hydrogel and disrupt the cellular membrane. Entrapment of viable cells within hydrogels, followed by lysis, could provide a convenient means for preparing biocatalysts without the need for enzyme extraction and purification. Hydrogel-immobilized cells are able to carry out chemical reactions within microfluidic channels. Specifically, a nonfluorescent dye, BCECF-AM, is able to penetrate both the hydrogel and the bacterial membrane and be converted into a fluorescent form (BCECF) by the interior cellular machinery. These results suggest that cells immobilized within microfluidic channels can act as sensors for small molecules and as bioreactors for carrying out reactions. PMID- 12530816 TI - Chemiluminescence detection for a microchip capillary electrophoresis system fabricated in poly(dimethylsiloxane). AB - Chemiluminescence (CL) detection integrated with a microchip capillary electrophoresis (MCE) system that was fabricated in poly(dimethylsiloxane) was demonstrated for chemical and biochemical analyses. Two model CL systems were involved here: metal ion-catalyzed luminol-peroxide reaction and dansyl species conjugated peroxalate-peroxide reaction. Different strategies based on three chip patterns (cross, cross combining with Y, and cross combining with V) to perform on-line CL detection for MCE were evaluated and compared in terms of sensitivity, reproducibility, and peak symmetry. The chip pattern of cross combining with Y proved to be promising for the luminol-peroxide CL system, while the chip pattern of cross combining with V was preferred for the peroxalate-peroxide system where CL reagent could not be effectively transported by electroosmotic flow. A detection limit down to submicromolar concentrations (midattomole) was achieved with good reproducibility and symmetric peak shape. Successful separation of three metal cations such as Cr(III), Co(II), and Cu(II) and chiral recognition of dansyl phenylalanine enantiomers within 1 min revealed distinct advantages of combining MCE with CL detection for rapid and sensitive analyses. PMID- 12530817 TI - Affinity capture and elution/electrospray ionization mass spectrometry assay of phosphomannomutase and phosphomannose isomerase for the multiplex analysis of congenital disorders of glycosylation types Ia and Ib. AB - We report a new application of affinity capture-elution electrospray mass spectrometry (ACESI-MS) to assay the enzymes phosphomannomutase (PMM) and phosphomannose isomerase (PMI), which when deficient cause congenital disorders of glycosylation CDG-type Ia and type Ib, respectively. The novel feature of this mass-spectrometry-based assay is that it allows one to distinguish and quantify enzymatic products that are isomeric with their substrates that are present simultaneously in complex mixtures, such as cultured human cell homogenates. This is achieved by coupled assays in which the PMM and PMI primary products are in vitro subjected to another enzymatic reaction with yeast transketolase that changes the mass of the products to be detected by mass spectrometry. The affinity purification procedure is fully automated, and the mass spectrometric analysis is multiplexed in a fashion that is suitable for high-throughput applications. PMID- 12530818 TI - Detection of pesticide residues on individual particles. AB - An aerosol time-of-flight mass spectrometer (ATOFMS) is used to analyze the size and composition of individual particles containing pesticides. Pesticide residues are found in the atmosphere as a result of spray drift, volatilization, and suspension of coated soils. The ability of the ATOFMS to identify the presence of these contaminants on individual particles is assessed for particles created from pure solutions of several commonly used pesticides, as well as pesticides mixed with an organic matrix, and coated on soils. The common names of the pesticides studied are 2,4-D, atrazine, chlorpyrifos, malathion, permethrin, and propoxur. Analysis of the mass spectra produced by single- and two-step laser desorption/ionization of pesticide-containing particles allows for identification of peaks that can be used for detection of pesticide residues in the ambient aerosol. The identified marker peaks are used to approximate detection limits for the pesticides applied to soils, which are on the order of a fraction of a monolayer for individual particles. Results suggest that this technique may be useful for studying the real-time partitioning and distribution of pesticides in the atmosphere immediately following application in agricultural regions. PMID- 12530819 TI - Dispersion in large aspect ratio microchannels for open-channel liquid chromatography. AB - Solute dispersion in open-channel liquid chromatography is often dominated by transverse diffusion limitations in the mobile phase (Martin, M.; Guiochon, G. Anal. Chem. 1984, 56, 614-620) convecting the solute species. While such dispersion is known to scale with the square of the Peclet number based on the narrower dimension of the conduit, the proportionality constant may significantly vary with the aspect ratio of the channel geometry. In this article, we investigate the effect of channel sidewalls on axial dispersion in electrokinetically and pressure-driven chromatographic systems. The analysis presented here clearly identifies the contribution from flow, wall retention, and the interaction between the two to the overall slug dispersion in the mobile phase for any arbitrary channel geometry. The particular geometries that have been investigated in this work, however, are the rectangular and the isotropically etched profiles often employed in microanalysis systems. Further, the effectiveness of simple double-etched profiles proposed elsewhere (Dutta, D.; Leighton, D. T. Anal. Chem. 2001, 73, 504-513) to diminish the effect of channel sidewalls on Taylor-Aris dispersion has also been examined. Analysis shows that dispersion arising due to shear and wall retention, as well as the interaction between the two, may be significantly reduced in large aspect ratio microchannels for optimized channel geometries. PMID- 12530820 TI - Comprehensive solid-phase extraction method for persistent organic pollutants. Validation and application to the analysis of persistent chlorinated pesticides. AB - The Centers for Disease Control and Prevention (CDC) is involved in many epidemiological studies regarding the measurement of chlorinated pesticides and polychlorinated biphenyls in specimens obtained from humans. In addition to these commonly determined analytes, there is a need to include additional persistent organic pollutants (POPs) in our analyses, which further stresses the analyses because sample volumes remain small. Thus, a single method of analysis for all POPs in human serum is needed. CDC has recently developed a semiautomated and comprehensive solid-phase extraction method for POPs. The method is comprehensive since it was optimized for the extraction of many different POP compound classes. We then developed a purification and fractionation scheme that allows (a) separation of different compound classes by particular functionalities and (b) purification of those fractions to remove coextracted interferences. This paper describes the first step in the semiautomated comprehensive extraction and multiple fractionation method developed by CDC for monitoring POPs. In this paper, we validate the analysis of the persistent chlorinated pesticides, a compound class difficult to examine because of their structural diversity, in human plasma. The method was validated against an existing CDC method by using a spiked quality-control serum pool. The concentrations determined for all analytes using both methods were within 2%-14% relative standard deviations. A multilevel (i.e., 3-4 point) matrix spike showed good linearity for the analytes tested (r2 = 0.978-0.999). The method was then applied to 40-year-old archived plasma samples for the quantitative analysis of selected chlorinated pesticides. Mean recoveries of the 13C-labeled internal quantification standards ranged from 64% to 123% for the 11 monitored pesticides. The overall method proved to be robust by handling old coagulated plasma samples. It allowed faster throughput of samples than our previous methods and provided cleaner samples with less frequent interferences or background as analyzed by high-resolution mass spectrometry. The method represents a preliminary step in establishing an automated, comprehensive multiresidue analysis method for POPs in human serum. PMID- 12530821 TI - Separation and quantitation of short-chain coenzyme A's in biological samples by capillary electrophoresis. AB - Because of the importance of coenzyme A's (CoA's or CoASH) in many metabolic processes and the biosynthesis of some carbohydrates and lipids, many methods have been developed to separate and determine their levels in various tissues for metabolism studies, including enzymatic assays, paper chromatography, and high performance liquid chromatography (HPLC). However, inadequate separation of coexisting CoA's in biological samples was often encountered due to the similarity of their structures. In this paper, we demonstrated for the first time the separation and quantitation of 12 different CoA's by using capillary electrophoresis with UV detection at 254 nm. All 12 CoA's (CoASH, HMG CoA, methylmalonyl CoA, succinyl CoA, methylcrotonyl CoA, isobutyryl CoA, oxidized CoA, acetyl CoA, crotonoyl CoA, n-propzoyl CoA, acetoacetyl CoA, malonyl CoA) were completely separated at -30 kV in a 100 mM NaH2PO4 running buffer containing 0.1% beta-cyclodextrin at pH 6.0. The total separation time was less than 30 min. The signal response was linear over 2 orders of magnitudes (from 1 to 100 nmol), and the detection limits were in the picomole range. The effects of pH, buffer concentration, additives, and operation voltages on sensitivity and resolution were also discussed. This technique, described here, is much more sensitive, faster, and simpler than the published HPLC methods and can potentially be used for mechanistic study in biological systems involving CoA metabolism. PMID- 12530822 TI - Competitive quenching fluorescence immunoassay for chlorophenols based on laser induced fluorescence detection in microdroplets. AB - An improved biomonitoring system for the analysis of 2,4,6-trichlorophenol (TCP) in urine samples has been developed. The principle of the biosensor device is the detection of laser-induced fluorescence (LIF) in single microdroplets by a homogeneous quenching fluorescence immunoassay (QFIA). The competitive immunoassay occurs in microdroplets (d = 58,4 microm) produced by a piezoelectric generator system with 10-microm-diameter orifice. A continuous Ar ion laser (488 nm) excites the fluorescent tracer; its fluorescence is detected by a spectrometer attached to a 512 x 512 cooled, charge-coupled device camera. Fluorescence is quenched by specific binding of TCP polyclonal antibodies to the fluorescent tracer (hapten A-fluorescein); the quenching effect is diminished by the presence of the analyte. Thus, an increase in the signal is produced in a positive dose-dependent manner when TCP is present in the sample. In 10 mM PBS buffer, the IC50 of the LIF-microdroplet QFIA is 0.45 microg L(-1) reaching a LOD of 0.04 microg L(-1). The QFIA with the same reagents performed in microtiter plate format achieved a LOD of 0.36 microg L(-1) in buffer solution. Performance in human urine was similar to that observed in the buffer. A LOD of 1.6 ,g L(-1), with a dynamic range between 4 and 149.5 microg L(-1) in urine, was obtained without any sample treatment other than dilution with the assay buffer. The detectability achieved is sufficient for occupational exposure risk assessment. PMID- 12530823 TI - Microenvironmental properties and chiral discrimination abilities of bile salt micelles by fluorescence probe technique. AB - The microenvironmental properties as well as the chiral discrimination abilities of four kinds of bile salt micelles were investigated by the fluorescence probe technique. A new fluorescence probe, 1,1'-bi-2-naphthol, was exploited to study the aggregation of bile salts, size, micropolarity, and microrigidity of the micelles with or without the presence of inorganic salts. Based on these results, the chiral discrimination abilities of bile salt micelles were further investigated by using (R)- and (S)-1,1'-bi-2-naphthol as chiral fluorescence probes. Different chiral discrimination ability was revealed by fluorescence spectra, fluorescence increase rate, and fluorescence quenching constants. The chiral discrimination mechanism of bile salt micelles was discussed. PMID- 12530824 TI - Headspace liquid-phase microextraction of chlorobenzenes in soil with gas chromatography-electron capture detection. AB - The organic solvent film formed in a microsyringe barrel was used as an extraction interface in headspace liquid-phase microextraction (HS-LPME) of chlorobenzenes. Some common organic solvents with different vapor pressures (9.33 12 918.9 Pa) were studied as extractants. The results indicated that even the solvent with the highest vapor pressure (cyclohexane) can be used to carry out the extraction successfully. In general, the reasons for successful extraction are the very small space (5 mm3) within the microsyringe barrel and the fast equilibrium between gaseous analytes and organic solvent film. Both of these factors significantly reduced the risk of solvent loss during extraction. Thus, the choice of extraction solvent for the present method was very flexible. From the viewpoint of extraction efficiency, toluene (which has relatively low vapor pressure) was found to provide the best extraction efficiency. The effects of sampling volume, organic solvent volume, syringe withdrawal rate, and number of extraction cycles were also investigated. The procedure with respect to repeatability and limits of detection was evaluated by soil spiked with chlorobenzenes. Repeatabilitywas between 5.7 and 17.7%, and the limits of detection were 6-14 ng/g. HS-LPME was shown to be an inexpensive, fast, and simple sample preparation method for volatile compounds. PMID- 12530825 TI - Combinational use of antibody affinities in an immunoassay for extension of dynamic range and detection of multiple analytes. AB - Here, we describe the coordinated use of two antibodies with different affinities in a single immunoassay to extend the dynamic range and to enable detection of multiple analytes. The combination of dual antibodies was permitted with a flow based assay at the antibody concentration below the dissociation constant, enabling affinity to govern the antibody-antigen binding. Both high and low affinity antibodies to estriol were used in combination to extend the range. The binding of each antibody was mutually independent and individually occurred over concentration ranges of 10 pM(-1) nM and 100 pM(-1) microM. The wide dynamic range of 10 pM(-1) microM was thus achieved as summation of the proportional signals to the total binding. When a combination of antibodies toward different antigens was used, it effectively detected multiple analytes within a mixture. In simultaneous analysis of a mixture of estradiol and estriol, the total signal was the sum of the binding signals from anti-estradiol and anti-estriol antibodies. In a further refinement, the individual antibodies were flowed through the flow cell sequentially, allowing the quantification of each binding signal within the combination. With this sequential format, measurement of the individual hormones in the range of 1.6 pM(-1) nM was shown. Furthermore, the same flow format was successfully applied to assay estriol and estradiol hormones in mixtures of six related compounds. PMID- 12530826 TI - A system for the direct determination of the nonvolatile organic carbon, dissolved organic carbon, and inorganic carbon in water samples through inductively coupled plasma atomic emission spectrometry. AB - A new system has been developed for the determination of total organic carbon (TOC) and inorganic carbon (IC) or total inorganic carbon (TIC) in waters. Only nonvolatile organic compounds can be detected through the present method. The system presented in this work is based on the measurement of the carbon atomic emission intensity in inductively coupled plasma atomic emission spectrometry (ICP-AES). This way, the organic matter does not undergo any preoxidation step. A semiautomatic accessory connected to the spectrometer separates the different carbon fractions (i.e., organic and inorganic). Because most of the solutions used in the present work did not contain suspended solid particles, the actual parameter that was determined was the dissolved organic carbon (DOC). The present system exhibits good sensitivities compared to those provided by conventional TOC and IC determination methods. The limits of detection obtained in the present work have been 0.07 and 0.0007 mg/L C in terms of TOC and IC, respectively. Furthermore, the system is able to handle high-salt-content solutions. This fact suggests that it would be possible to analyze seawater samples, avoiding some of the problems encountered with conventional methods, such as system blocking or interferences. The TOC and IC values found for natural samples are very close to those measured using conventional methods. The ICP-AES method has been successfully used in two interesting applications: (i) monitoring the efficiency of a water treatment plant and (ii) determining the contents of dissolved carbon dioxide, on one hand, and that of carbonate and bicarbonate, on the other, in the same sample. PMID- 12530827 TI - On-line overpressure thin-layer chromatographic separation and electrospray mass spectrometric detection of glycolipids. AB - On-line thin-layer chromatographic separation and electrospray mass spectrometry (TLC/ESI-MS) has been accomplished by direct linking of a commercial overpressure TLC instrument, OPLC 50, and a Q-TOF mass spectrometer. Mass spectrometric detection sensitivity and chromatographic resolution achieved by this configuration were assessed using acidic glycolipids as examples. Under the optimized conditions, a sensitivity of 5 pmol of glycosphingolipid was readily demonstrated for TLC/ESI-MS and 20 pmol for TLC/ESI-MS/MS production scanning to derive the saccharide sequence and long chain base/fatty acid composition of the ceramide. Initial preconditioning of TLC plates is necessary to achieve high sensitivity detection by reducing chemical background noise. Plates can be used repeatedly (at least 10 times) for analysis, although this may result in a minor reduction in TLC resolution. Following solvent development, separated components on the TLC plates can be detected in the conventional way by nondestructive staining or UV absorption or fluorescence and can be stored for on-line TLC/ESI MS analysis at a later stage without reduction in mass spectrometric detection sensitivity and chromatographic resolution. Aspects for further improvement of OPLC instrumentation include use of narrower TLC plate dimensions and refined design of the eluate exit system. PMID- 12530828 TI - A micromachined calorimetric gas sensor: an application of electrodeposited nanostructured palladium for the detection of combustible gases. AB - Palladium films with regular nanoarchitectures were electrochemically deposited from the hexagonal (H1) lyotropic liquid crystalline phase of the nonionic surfactant octaethyleneglycol monohexadecyl ether (C16EO8) onto micromachined silicon hotplate structures. The H1-e Pd films were shown to have high surface areas (approximately 28 m2 g(-1)) and to act as effective and stable catalysts for the detection of methane in air on heating to 500 degrees C. The response of the H1-e Pd-coated planar pellistors was found to be linearly proportional to the concentration of methane between 0 and 2.5% in air with a detection limit below 0.125%. Our results show that the electrochemical deposition of nanostructured metal films offers a promising approach to the fabrication of micromachined calorimetric gas sensors for combustible gases. PMID- 12530829 TI - Mechanistic insights into the development of optical chloride sensors based on the [9]mercuracarborand-3 ionophore. AB - Fluorescent sensing microspheres based on perhaps the most selective and practically useful chloride ionophore known, the recently reported [9]mercuracarborand-3 (MC-3), have been prepared and optimized for physiological measurements. In initial work, this ionophore was shown to yield functional optical sensing films in combination with an electrically neutral chromoionophore, ETH 5418. Unfortunately, however, these optodes suffered from unacceptably high levels of sodium interference under physiological conditions. To better understand the sensing mechanism, optical and potentiometric binding experiments were used to characterize the stoichiometry and the complex formation constants for this ionophore. It was found that the preferred stoichiometry is 1:2, rather than 1:1 as assumed earlier. The 1:2 complex is extremely stable (logbeta2 = 13.4), but a relatively strong 1:1 complex also exists (log K1 = 9.9). These characteristics were used to fabricate chloride optodes that make use of the stepwise ion-ionophore decomplexation equilibrium, by adding a calculated amount of lipophilic anion exchanger to the polymer film. Such optodes showed dramatically reduced sodium interference while maintaining the excellent selectivity of the traditional formulation. The optimized composition also shifted the measuring range to physiological conditions, making them useful for the assessment of chloride in undiluted and 10-fold-diluted blood at pH 7.4. After necessary alterations of the particle preparation procedure and sensor formulation, the new insights were used to fabricate mass-produced optical sensing microspheres with characteristics essentially identical to those of the optode sensing films. PMID- 12530830 TI - All-solid-state calcium solvent polymeric membrane electrode for low-level concentration measurements. AB - An all-solid-state calcium-selective electrode with a plastic membrane phase containing a calcium ionophore ETH-1001 placed on poly(3-methylthiophene) ion-to electron transducting layer functionalized to bind calcium cations has been constructed. The obtained potentiometric sensors were characterized with a calibration line of slope close to Nernstian within the activity from 10(-5) to 0.1 M. For Ca2+ activity lower than 10(-5) M, super-Nernstian behavior was observed. The super-Nernstian response that is observed for electrodes with an internal solution strongly binding primary ions was in this case attributed to incorporation of calcium ions in the modified solid-contact phase. With this arrangement, the evaluation of selectivity coefficients much closer to those that depend on the properties of the ion-selective membrane itself was possible. PMID- 12530831 TI - Detection of ATP-induced nitric oxide in a biomimetic circulatory vessel containing an immobilized endothelium. AB - Conditions for the adhesion of bovine pulmonary artery endothelial cells (bPAECs) in microbore tubing of 250-microm i.d. are described. When immobilized to the lumen of microbore tubing, these cells represent a mimic of a circulatory vessel's endothelium. The microbore tubing is coated with 100 microg mL(-1) fibronectin in order to promote bPAEC adhesion to the lumen of the tubing. A series of micrographs of the cells inside of the tubing indicates that approximately 3.5 h is necessary for cell adhesion. In this study, adenosine triphosphate (ATP) is used to induce the release of nitric oxide from the endothelium mimic. The endothelium-derived NO is detected amperometrically at a parallel flow cell containing a glassy carbon working electrode modified with Nafion. Results indicate that detectable amounts of NO are only produced by the endothelium mimic when ATP is present in the buffer. The typical concentration of NO produced by the endothelium mimic upon the introduction of 100 microM ATP is approximately 0.80 microM. Based on the injection volume of ATP and the estimated number of cells on the tubing lumen, this value corresponds to approximately 1 amol of NO/cell. Moreover, shear stress alone does not provide the agonistic effect required for NO production in the submicromolar range. PMID- 12530832 TI - Synthesis of an ammonium ionophore and its application in a planar ion-selective electrode. AB - A modular technique was used to synthesize an ammonium-selective ionophore based on a cyclic depsipeptide structure. The ionophore was incorporated into a planar ion-selective electrode sensor format and the selectivity tested versus a range of metal cations in a commercial clinical diagnostic "point-of-care" instrument. Four sensor membrane formulations were tested, all of which consisted of plasticized PVC. Formulations differed as to the type of plasticizer used and whether an ionic additive was present. It was found that the membrane containing the polar plasticizer nitrophenyl octyl ether in the absence of ionic additive exhibited near-Nernstian behavior (slope, 60.1 mV/decade at 37 degrees C) and possessed high selectivity for ammonium ion over lithium and the divalent cations, calcium and magnesium (log K(POT)NH4+(j) = -7.3, -4.4, and -7.1 for lithium, calcium, and magnesium ions, respectively). The same membrane also exhibited sodium and potassium selectivity that was comparable to that reported for nonactin (log K(POT)NH4+(j) = -2.1 and -0.6 for sodium and potassium, respectively, compared to -2.4 and -0.9 in the case of nonactin). Membranes containing the less polar plasticizer, dioctyl phthalate, showed sub-Nernstian behavior (slope, <50 mV/decade at 37 degrees C). In all cases, the presence of the ionic additive potassium tetrakis(4-chlorophenyl)borate substantially reduced the selectivity observed. The flexible modular synthetic technique developed and reported here will allow the cyclic depsipeptide structure to be tuned for optimum selectivity. PMID- 12530833 TI - Ultrasensitive detection of unstained proteins in acrylamide gels by native UV fluorescence. AB - Visualization of proteins inside acrylamide and other gels usually relies on different staining methods. To omit the protein-staining procedure, we visualized unstained proteins inside acrylamide gels by laser excitation with ultraviolet (UV) light (280 nm, 35 mJ/cm2) and directly detected native UV fluorescence. In one-dimensional gels, a detection limit as low as 1 ng for bovine serum albumin and 5 ng for other proteins with a linear dynamic range (2.7 orders of magnitude) comparable to state of the art fluorescent dyes could be achieved. In addition, the application of this method to 20 microg of a whole cell lysate separated in a two-dimensional gel showed more than 600 spots. Since protein labeling always represents a serious obstacle in protein identification technologies, the working efficiency with our procedure can be considered as a significant improvement for protein visualization and reproducibility in proteomics. PMID- 12530834 TI - Infrared laser isolation of ions in Fourier transform mass spectrometry. AB - A new method for isolating ions for tandem mass spectrometry analyses in Fourier transform mass spectrometry is illustrated. The method employs an infrared laser to dissociate completely the undesired ions. The selected ions are excited to an orbit away from the degradative portion of the laser beam. Ion isolation was accomplished and tandem mass spectrometry experiments were performed on model oligosaccharides and compounds from biological samples. PMID- 12530835 TI - Developing ergonomic solutions for prevention of musculoskeletal disorder disability. AB - Numerous models have been proposed to help understand the relationship between personal and work factors that affect participation in work. The authors use a generic version of these models as a framework for identifying gaps between job demands and worker capacities. They describe metrics for assessing factors associated with causing or aggravating musculoskeletal disorders. Two case examples are presented to illustrate the hierarchical assessment of jobs and the evaluation of gaps between job demands and worker capacities. Finally, the authors describe the development of a job database to facilitate future evaluations. PMID- 12530836 TI - Ergonomic job design to accommodate and prevent musculoskeletal disabilities. AB - Work-related musculoskeletal disorders (MSDs) account for a major portion of the cost of work-related injury and illness in the United States. Many of these injuries and illnesses lead to temporary or permanent disability. It is generally accepted that the incidence of MSDs increases when the demands of the job exceed the capabilities of the worker. As the workforce ages and physical capabilities decline, it is anticipated that many more Americans will request disability related leave resulting from musculoskeletal disorders because they are unable to meet the demands of the job. To prevent these disabilities and to accommodate a wider range of people in the workforce, physical job demands may have to be reduced so that a larger portion of the population will be capable of working. Providing engineering controls or alternative work arrangements allows for accommodation of workers with a wide range of capabilities and can assist in rehabilitation and early return to work following injury. PMID- 12530837 TI - Clinical tools to facilitate workplace accommodation after treatment for an upper extremity disorder. AB - Failure to implement work site accommodations for work-related upper extremity disorders (WRUEDs) may be a factor contributing to delayed functional recovery and relapse. The present study describes the use of the 38-item Job Requirements and Physical Demands (JRPD) scale, a self-report measure of ergonomic exposure, and other case management tools to improve accommodation efforts for 101 workers (75 women, 26 men) returning to work after lost time related to a WRUED. Items were categorized into five subscales based on item content: administrative, computer-related, workstation design, environmental, and equipment. Administrative risk factors were elevated among office clerks, whereas postal clerks and letter carriers reported more workstation design risk factors, and letter carriers and electrical/mechanical workers cited more equipment-related risk factors (p < 0.05). All occupational categories rated computer-related risk factors highest. The Integrated Case Management (ICM) approach, which relies on the JRPD scale to guide recommendations, was used with a subgroup of these workers (n = 53), resulting in 1.4 times more workplace accommodations per worker than with a non-ICM approach. Clinical use of the self-reported exposure measure within the overall workplace accommodation process may have been a factor contributing to more frequent accommodation in the ICM group. This study of a subgroup of workers' compensation cases highlights the need for additional investigation of tools to integrate ergonomic approaches within the workplace accommodation process. PMID- 12530838 TI - Remote measurement methods for user-centered design of telesupport systems. AB - The term telesupport is broad and encompasses many different applications with the shared mission of relying on electronic means to deliver services that improve quality of life. However, telesupport systems lack empirical research and efficient methods for the development of user requirements and design guidelines. Remote assessment methods using focus groups, as well as a suite of remote usability metrics using critical incidents, are described. These can be used to develop human factors design principles suitable for implementation in telesupport systems and other assistive technologies used by a broad range of people with disabilities. An integrated measurement system and extensions to remote measurement methods are discussed as a means of improving user-centered design of telesupport systems. PMID- 12530839 TI - User performance with speech recognition: a literature review. AB - The application of speech recognition to the computer access needs of people with disabilities continues to grow, and a greater understanding of user performance with such systems is needed. This article reviews what is known about user performance with speech recognition systems, with a focus on its application to accommodation of physical disability. Although current systems offer the potential of text entry at 150 words per minute, the literature suggests that users actually achieve somewhere between 8 and 30 words per minute. Barriers that may contribute to this gap, such as the costs associated with correcting recognition errors, are reviewed, and directions for future research are proposed. A major need is for additional research involving users who have physical disabilities. PMID- 12530840 TI - Employer attitudes, training, and return-to-work outcomes: a pilot study. AB - This work was concerned with investigating and changing employer attitudes and practices that impede return to work and rehabilitation for injured workers. Prior studies have shown that employer responses to workers reporting work related musculoskeletal discomfort have significant and independent effects on disability outcomes. Based on these findings, a pilot training program was developed by occupational rehabilitation specialists to improve the response of supervisors to employees reporting work-related injuries. The training was delivered to 108 supervisors at seven southeastern New Hampshire companies that volunteered to participate. A survey was developed and pilot-tested, then administered before the training. Survey participants were employees who had work related injuries in the past year. The survey asked about types and onset of injury, specific supervisor responses, and overall impression of supervisor interaction at the time of injury. Employee responses before the intervention were compared with those collected from workers who reported injuries after the training. The postintervention results demonstrated significant decreases in supervisors (1) blaming employees for the injury, (2) not taking the condition seriously, and (3) discouraging the worker from filing a claim. Positive trends in confidentiality of discussions, access to medical care, and accommodation and work modifications were also noted. Although anecdotal reports from the companies indicated a consistent decrease in work-related lost time after the intervention, actual verification was not possible, and other components of the intervention may have accounted for this outcome. Small numbers of cases and possible lack of comparability of cases before and after the intervention are significant limitations. However, rehabilitation professionals may be able to improve disability management practices and accommodations through employer education, especially when training is directed toward front-line supervisors. PMID- 12530841 TI - Fullerene-coated beads as reusable catalysts. AB - New heterogeneous catalysts that use oxygen and light to generate singlet oxygen ((1)O2) have been prepared. The catalysts facilitate various types of singlet oxygenation reactions including the Ene reaction, the Diels-Alder reaction, and others. The catalysts are made by stirring a heterogeneous mixture of fullerene C(60) (dissolved in toluene) with aminomethylated poly(styrene-co-divinylbenzene) beads. Also, catalysts for aqueous photooxidations are made by reacting the initial catalysts with poly(allylamine) to create an outer layer that is more hydrophilic. PMID- 12530842 TI - Solution structure of the antitumor candidate trunkamide A by 2D NMR and restrained simulated annealing methods. AB - Trunkamide A (1) is a cyclic heptapeptide extracted from the ascidian Lissoclinum sp. and has shown very promising cytotoxic activity. This compound incorporates several of the motifs commonly observed in the Patellin family, including dimethylallyl (Dma) Thr and Ser side chains and a thiazoline heterocycle. Given that little is known about the structures adopted by the cyclopeptides of the Patellin family, and with the aim of establishing structure-activity relationships, we have carried out the conformational analysis of trunkamide A by a combination of 2D NMR experiments and simulated annealing calculations. Our results show that the conformation of 1 is very rigid and is dominated by the volume of the dimethylallyl side chains and two trans-annular hydrogen bonds. We have also studied the conformation of 2, the l-Phe diastereoisomer of 1, the analysis of which provides a possible rationale for its epimerization to 1, a process that is observed in solution. Finally, we show how a thorough NMR characterization can be used, in combination with simulated annealing methods, to confirm the configuration of a stereogenic center in the backbone of a rigid cyclic peptide such as trunkamide A (1). PMID- 12530843 TI - Stable simple enols. Resolution of chiral 1-[9'-(2'-fluoroanthryl)]-2,2 dimesitylethenol. A different racemization mechanism for the enol and its acetate. AB - Chiral 1-[9'-(2'-methoxyanthryl)]-2,2-dimesitylethenol (2), 1-[9'-(2' fluoroanthryl)]-2,2-dimesitylethenol (3), and 1-[9'-(2'-fluoroanthryl)]-2,2- dimesitylvinyl acetate (4) were synthesized and their DNMR behavior in C6D5NO2 was studied. 3 and 4 were resolved on an amylose tris(3,5 dimethylphenylcarbamate) HPLC column to their enantiomers. Acetate 4 racemizes slowly in solution with DeltaG(e)(++), DeltaH(e)(++), and DeltaS(e)(++) values of 26.2, 27.6 kcal mol(-)(1), and 4.3 eu, respectively, as expected for a rotational betabeta'-2-ring flip process in a vinyl propeller and the racemization is unaffected by added TFA, Et3N, and EtOD. Although 3 racemizes almost 350 times faster, the racemization is catalyzed by TFA and shows bell-shape catalysis by Et3N and a KIE in a partially deuteriated solvent. From this and the DNMR data, it is concluded that 3 does not racemize via a rotational betabeta'-2-ring flip. Five nonflip routes are discussed for the racemization of 3, and it is concluded that only the one initiated by protonation at C1 does not contradict the experimental data. By analogy with the E/Z isomerization of the structurally related 2-(m-methoxymesityl)-1,2-dimesitylethenol 17, it is suggested that in the absence of added catalyst one or more enol molecule(s) catalyze the enantiomerization of another one. Only partial resolution was achieved for 2 and from the similarity of its behavior with that of 3, it is suggested that it racemizes by the same mechanism. PMID- 12530844 TI - Synthesis of furo[3,4-c]furans using a rhodium(II)-catalyzed cyclization/Diels Alder cycloaddition sequence. AB - A series of 2-alkynyl 2-diazo-3-oxobutanoates, when treated with a catalytic quantity of rhodium(II) acetate, afforded furo[3,4-c]furans in good yield. The reaction proceeds by addition of a rhodium-stabilized carbenoid onto the acetylenic pi-bond to give a vinyl carbenoid that subsequently cyclizes onto the neighboring carbonyl group to produce the furan ring. These furo[3,4-c]furans react with various dienophiles, furnishing anisole derivatives derived by loss of water from the initially formed Diels-Alder cycloadducts. The Rh(II)-catalyzed cyclization reaction was quite versatile with regard to the nature of the interacting carbonyl group. The methodology was applied to the synthesis of several oxa-polyheterocyclic systems by first generating a 2-alkoxy-substituted furan and then allowing it to undergo a subsequent intramolecular Diels-Alder cycloaddition. Ring opening of the resulting cycloadduct is followed by deprotonation to furnish a rearranged keto lactone. The potential use of this method for the synthesis of the alkaloid strychnine was probed using suitable model diazo compounds. To establish the viability of this approach, the Rh(II) catalyzed cyclization/cycloaddition sequence of alpha-diazo amides 64 and 68 were studied. Both compounds underwent the sequential process in good overall yield, leading to novel pentacyclic products. The structural features of the resultant products present numerous opportunities for postcycloaddition manipulations that could be exploited to synthetic advantage. PMID- 12530845 TI - Molecular tectonics. Construction of porous hydrogen-bonded networks from bisketals of pentaerythritol. AB - 2,4,8,10-Tetraoxaspiro[5,5]undecanes tetrasubstituted at the 3 and 9 positions with groups incorporating diaminotriazines can be used for the construction of extensively hydrogen-bonded networks by the strategy of molecular tectonics. Four such compounds, tectons 1-4, were made by short and efficient syntheses involving bisketalization of pentaerythritol and subsequent reactions. Unlike tectons typically used in previous studies, compounds 1-4 are flexible and chiral, and they orient four sticky diaminotriazine groups in a distorted tetrahedral geometry. Tecton 1 crystallizes from DMF/toluene as an inclusion compound of approximate composition 1.8DMF.xH2O. In the resulting structure, each tecton participates in a total of 16 hydrogen bonds. Eight of these bonds involve four principal neighbors, and the tectons linked in this way define a distorted diamondoid network. Despite 8-fold interpenetration, 60% of the volume of the network is available for including guests. The guests are disordered and occupy parallel helical channels that have cross sections of approximately 11 x 12 A2 at the narrowest points. These channels provide access to the interior of the crystals and permit guests to be exchanged quantitatively without loss of crystallinity. It is noteworthy that tecton 1, despite its flexibility, small size, and structural simplicity, is apparently unable to find a periodic three dimensional structure in which the dictates of hydrogen bonding and close packing are satisfied simultaneously. PMID- 12530846 TI - Efficient synthesis of porphyrin-containing, benzoquinone-terminated, rigid polyphenylene dendrimers. AB - A series of rigid polyphenylene, free-base porphyrin-containing dendrimers terminated with either dimethoxybenzene or benzoquinone end-groups were prepared by a combined divergent and convergent synthesis. Unlike previous routes for preparing polyphenylene dendrimers that are incompatible with end-groups bearing certain functional moieties, the synthetic methodology chosen for this work enables incorporation of functional groups on the dendrimer end-groups during preparation of the dendrimer wedges and during synthesis of the final dendrimer. The basic strategy utilized a convergent preparation of dendrimer wedges using Suzuki coupling conditions, which were then either attached to a porphyrin core in a divergent coupling step or cyclized to form the porphyrin dendrimer in a convergent step. The latter approach was found to be more general and resulted in higher yields and more readily separated products. Steady-state absorption measurements for these dendrimers showed Soret and Q-band absorptions typical of free-base porphyrins. Preliminary steady-state fluorescence measurements of these dendrimers indicate quenching of the S1 state of the free-base porphyrin in all benzoquinone-containing dendrimers that is attributed to efficient electron transfer from the excited porphyrin to the benzoquinone end-groups. The amount of fluorescence quenching was in good agreement with the number of benzoquinone groups at the dendrimer periphery and the distance between the porphyrin and benzoquinone groups as calculated by semiempirical (AM1) molecular orbital calculations. PMID- 12530847 TI - Remote substituent effects on N-X (X = H, F, Cl, CH3, Li) bond dissociation energies in para-substituted anilines. AB - UB3LYP/6-311++g**//UB3LYP/6-31+g* and ROMP2/6-311++g**//UB3LYP/6-31+g* methods were used to calculate (i) N-X bond dissociation energies (BDE) in 4-YC6H4NH-X and (ii) N-H BDEs in 4-YC6H4NU-H, where Y = H, Me, OCH3, SMe, NH2, NMe2, SiMe3, F, Cl, CN, COOH, CF3, and NO2, X = H, CH3, F, Cl, and Li, and U = H, F, and CH(3). It was found that N-H BDEs of 4-YC6H4NH2 have a positive correlation with the substituent sigma(p+) constants. The slope (rho+) is about 3.0-4.3 kcal/mol, which is in good agreement with the experimental results. It was also found that the substituent effects on N-X BDEs of 4-YC6H4NH-X change considerably when X changes. rho(+)values for N-CH3, N-F, N-Cl, and N-Li BDEs were calculated to be 3.1-4.6, 1.3-1.9, 1.8-2.6, and 4.9-6.8 kcal/mol, respectively. The reason for the variation of substituent effects was proposed to be the ground-state effect, i.e., the interaction between the intact NH-X moiety and the parasubstituents. Finally, alpha-substitution was found to be able to significantly change the substituent effects. rho(+)values for N-H BDEs of 4-C6H4NCH3(-)H and 4-C6H4NF-H are 2.5-4.0 and 1.7-1.9 kcal/mol, respectively. PMID- 12530848 TI - Electrophilic tetraalkylammonium nitrate nitration. II. Improved anhydrous aromatic and heteroaromatic mononitration with tetramethylammonium nitrate and triflic anhydride, including selected microwave examples. AB - A new one-pot nitration employing tetramethylammonium nitrate and trifluoromethanesulfonic anhydride in dichloromethane to provide a ready source of the nitronium triflate nitrating agent is presented. Rapid and selective nitration with a variety of aromatic and heteroaromatic substrates is achieved resulting in the synthesis of several novel organic compounds. A distinct advantage is the removal of undesired byproducts by aqueous workup. This very mild nitration permits large-scale syntheses and gives high isolated product yields that often require no further purification. This tetramethylammonium nitrate-based nitration also has been applied to microwave-assisted conditions, and the results with several compounds are outlined. PMID- 12530849 TI - Efficient synthesis of 3H-imidazo[4,5-b]pyridines from malononitrile and 5-amino 4-(cyanoformimidoyl)imidazoles. AB - 1-Aryl-5-amino-4-(cyanoformimidoyl)imidazoles 2 were reacted with malononitrile under mild experimental conditions and led to 3-aryl-5,7-diamino-6-cyano-3H imidazo[4,5-b]pyridines 5, when the reaction was carried out in the presence of DBU, or to 3-aryl-5-amino-6,7-dicyano-3H-imidazo[4,5-b]pyridines 3, in its absence. Both reactions evolved from the adduct formed by nucleophilic attack of the malononitrile anion to the carbon of the cyanoformimidoyl substituent. A 5 amino-1-aryl-4-(1-amino-2,2-dicyanovinyl)imidazole 4 was isolated when this reaction was carried out in the presence of DBU. The structure of compound 4 was confirmed by spectroscopic methods and by reaction with triethyl orthoformate and with acetic anhydride, leading respectively to 9-aryl-6-(cyanomethylidene)purines 11 and 12. Imidazole 2b was also reacted with ethyl acetoacetate, a carbon acid with a pK(a) comparable to that of malononitrile. Similar reaction conditions were used and the product isolated was a 6-carbamoyl-1,2-dihydropurine 10, showing that a different mechanism was operating in this case. PMID- 12530850 TI - Solution and crystal lattice effects on the photochemistry of 6-substituted cyclohexenones(1)(,)(2). AB - The photochemistry of 13 4,4-diphenylcyclohexenones, substituted at carbon-6, was investigated in solution and in the crystalline state. The stereoselectivity was of particular interest. In the solution photochemistry of C-6 monosubstituted enones in benzene, there was a unique preference for migration of the cis-phenyl group with formation of bicyclo[3.1.0]hexanone photoproducts, with the original 6 substituent having an endo configuration at carbon-3 of the product. In methanol the reaction was diverted to afford 3,4-diphenylcyclohex-2-enes understood as arising from a hydrogen-bonded zwitterionic intermediate. The solid-state photochemistry was also investigated. There was a dramatic absence of the 3,4 diphenylcyclohex-2-ene products in accord with the absence of the hydrogen bonding encountered in methanol. Further, the solid-state reactivity correlated with a vector analysis using X-ray atomic coordinates. This established that the migrating phenyl group required an orientation facing the enone beta-carbon. While the interesting preference for the cis-endo migration was not intuitively predicted, ab initio computations on the alternative phenyl-bridged triplet intermediates did lead to an understanding of the selectivity. PMID- 12530851 TI - Haouamines A and B: a new class of alkaloids from the ascidian Aplidium haouarianum. AB - The chemical study of the ascidian Aplidium haouarianum has led to the isolation of the new metabolites haouamines A (1) and B (2) which belong to a novel class of alkaloids. The structure of 1 was established by interpretation of its spectroscopic data and those of the N-methyl derivative 3, and confirmed by X-ray crystallographic analysis. The structure of 2 was deduced by spectroscopic study of its peracetyl derivative 2a. In solution each haouamine exists as an unseparable mixture of two interconverting isomers derived by the presence of a highly strained 3-aza-[7]-paracyclophane moiety in their structures. Compound 1 exhibits a selective cytotoxic activity against the HT-29 human colon carcinoma cell line. PMID- 12530852 TI - Daphcalycine, a novel heptacycle fused ring system alkaloid from Daphniphyllum calycinum. AB - A novel Daphniphyllum alkaloid, daphcalycine (1), was isolated together with known daphnicyclidin D (2) from the stem bark of Daphniphyllum calycinum. The highly condensed polycyclic structure, established by spectral analysis, possessed an unusual framework: a central quinuclidine like tricycle produced by fusion of a piperidine, a tetrahydropyran, and an oxazine ring in turn condensed to surrounding three penta-, one hexa-, and one hepta-membered rings. The relative configuration of 11 carbon stereocenters of 1 was elucidated on the basis of NOESY. PMID- 12530853 TI - Aromatic bromination versus oxidation of indolylmalonates by bromine. AB - The reactions of 5-substituted indolylmalonates (2a-e), carrying an electron withdrawing group at the N(1) position, with bromine in CCl(4) or AcOH are reported. These substrates undergo oxidation in competition with the well-known aromatic bromination. Under the two sets of conditions, with parent indolylmalonate (2a), chemospecific oxidation is observed, whereas with 5 hydroxyindolylmalonate (2c), bromination at the 4- and 6-position is the dominating reaction. Investigation of the products composition of several 5 substituted indolylmalonates revealed the following trend: with a 5-substituted electron-withdrawing group like fluorine, the indolylmalonate undergoes oxidation rather than bromination. In contrast, with a 5-substituted electron-donating group, like a hydroxyl group, the ring bromination occurs preferentially over the oxidation. When the 5-substituent is an alkoxyl group, a significant amount of brominated-oxidized products is obtained. Monitoring the oxidation reaction by mass spectrometry allowed the characterization of the 2-bromoindolylidenemalonate intermediate. A bromonium ion is considered as possible pathway in the formation of this intermediate. The conformation of unsymmetrical methoxyl and benzyloxyl substituents was determined from (1)H NMR spectra, single-crystal X-ray diffraction and ab initio calculations. PMID- 12530855 TI - Tuning the optical properties of fluorinated indolylfulgimides. AB - Photochromic fluorinated indolylfulgides have been identified as potential candidates for a wide range of applications including optical switches, photoregulators of biological processes, and optical memory media. In humid environments or biological systems, hydrolytic stability is essential. In an effort to improve hydrolytic stability, a series of indolylfulgimides has been synthesized from a parent trifluoromethyl-substituted indolylfulgide. The nitrogen of the succinimide moiety is linked to either a dimethyl amino or one of seven substituted phenyl groups. The phenyl groups feature substituents with increasing electron-withdrawing ability. The spectral characteristics of each compound have been examined, revealing that the wavelength absorption maxima of each form increases with increasing electron-withdrawing ability of the substituted N-phenyl ring. The quantum yields of the photoreactions have been determined with the N-(phenyl)fulgimide showing a ring closure value of nearly 0.30 in toluene. In addition, the hydrolytic, thermal, and photochemical stabilities of each compound have been measured. The fulgimides exhibit at least a 200-fold enhancement of hydrolytic stability for the Z-form and over a 1000 fold enhancement for the C-form in comparison to the same form of the parent fulgide. The N-(2,3,5,6-tetrafluoro-4-trifluoromethylphenyl)fulgimide can undergo up to 3000 photochemical cycles (coloration followed by bleaching) before losing 20% of its initial absorbance at photostationary state. PMID- 12530854 TI - Mn(III)/Cu(II)-mediated oxidative radical cyclization of alpha (Methylthio)acetamides leading to erythrinanes. AB - Treatment of N-[2-(3,4-dimethoxyphenyl)ethyl]-alpha-(methylthio)acetamide 3 with Mn(OAc)3 in the presence of Cu(OAc)2 gave tetrahydroindol-2-one 4, which then cyclized with Mn(OAc)3 to give 4-acetoxyerythrinane 5. A similar reaction of the 3,4-methylenedioxyphenyl congener 8 also gave tetrahydroindol-2-one 9, which, however, gave only a trace amount of the Mn(OAc)3-mediated cyclization product 11 and afforded the oxidation product 10. On the basis of these results, formation of 5 from 4 was thought to proceed via nucleophilic attack of the pyrrole ring on the cation-radical lX, generated by a single electron-transfer reaction of the acetoxy-substituted intermediate V. Treatment of compound 16 with Mn(OAc)3/Cu(OAc)2 gave no erythrinane derivative with recovery of the starting material, indicating that the presence of a methylthio group of 4 is essential for effecting the formation of erythrinane 5. On the other hand, treatment of 3 with Mn(OAc)3 using Cu(OTf)2 as an additive in place of Cu(OAc)2 gave another erythrinane 17. This method was applied to a formal synthesis of 3 demethoxyerythratidinone (20), a naturally occurring Erythrina alkaloid. PMID- 12530856 TI - Solid-state diphotocyclization of iso- and terephthalaldehydes via dihalogen substitution. AB - The supramolecular nonbonded C-H...X interactions between formyl hydrogens and ortho-halogen atoms (Br/Cl) have been exploited to achieve conformational control in the solid state of dimethyl-substituted iso- and terephthaladehydes (1-3) for unprecedented diphotocyclization. It is shown that the dihalogen substitution also contributes to the stability of the benzocyclobutenols relative to their precursor photoenols, so that the solid-state photolysis of dialdehydes 2b, 2c, and 3b leads to diphotocyclization to afford respectable yields of bis benzocyclobutenols. PMID- 12530857 TI - Artificial nucleosides possessing metal binding sites at the 3'- and 5'-positions of the deoxyribose moieties. AB - This paper describes a convenient synthetic procedure for nucleoside mimics, 1-6, in which the 3',5'-hydroxy groups of natural 2'-deoxythymidine or 2' deoxyadenosine are replaced by thiol, amine, or alkylthiol groups. Such nucleosides would be built up into a single DNA strand with cooperative participation of metal coordination, where internucleoside linkages are replaced by metal complexation motifs. The X-ray crystal structure and complexation behaviors of 3',5'-dithiothymidine, 1, with Au(I) are also reported. PMID- 12530858 TI - Carbopalladation of nitriles: synthesis of 3,4-disubstituted 2-aminonaphthalenes and 1,3-benzoxazine derivatives by the palladium-catalyzed annulation of alkynes by (2-Iodophenyl)acetonitrile. AB - Intramolecular carbopalladation of the cyano group has been employed for the synthesis of 3,4-disubstituted 2-aminonaphthalenes. (2-Iodophenyl)acetonitrile reacts with a variety of internal alkynes to afford 2-aminonaphthalenes in high yields with good regioselectivity. The scope and limitations of this process, which proceeds by the intramolecular addition of a vinylpalladium species to the triple bond of the cyano group, have been studied. The annulation of certain hindered propargylic alcohols affords 1,3-benzoxazine derivatives, rather than the expected 2-aminonaphthalenes. The involvement of trialkylamine bases in the formation of these heterocyclic compounds has been established. A proposed mechanism for the synthesis of 1,3-benzoxazine derivatives involves the formation of the expected 2-amino-3-(1-hydroxyalkyl)naphthalenes, followed by their condensation with an iminium ion species formed from the trialkylamine base used in the reaction. PMID- 12530859 TI - First asymmetric synthesis of 6-hydroxy-4-sphingenine-containing ceramides. Use of chiral propargylic alcohols to prepare a lipid found in human skin. AB - 6-Hydroxy-(4E)-sphingenine-containing ceramides were found recently in human skin. We present here the first synthesis of the 6S and 6R diastereoisomers 2 and 3, which represent analogues of (2S,3R)-ceramide (1) having two allylic hydroxyl groups. Chiral propargylic alcohols 8 and 11, which were prepared by asymmetric dihydroxylation of alpha,beta-unsaturated ester 13 and allylic chloride 22, respectively, were employed as precursors of 2 and 3. Nucleophilic addition of lithiated TBS-protected propargylic ethers 25 and 32 to l-serine-derived aldehyde 26, respectively, afforded oxazolidine intermediates 27 and 33. Acid-mediated deprotection of the oxazolidine, followed by N-acylation and Birch reduction, completed the syntheses of 2 and 3. PMID- 12530861 TI - High-intensity ultrasound-promoted reformatsky reactions. AB - Reformatsky reactions of a phenyl ketone, an alpha-bromoester, zinc dust, and a catalytic amount of iodine in dioxane under high-intensity ultrasound (HIU) irradiation from an ultrasonic probe give high yields of beta-hydroxyesters in short reaction times. A series of alkyl phenyl ketones with increasing steric demands of the alkyl group are evaluated as potential electrophiles for the reactions with several alpha-bromoesters, also having increasing steric demands. The Reformatsky reaction under HIU is found to be concentration dependent. PMID- 12530860 TI - Synthesis and growth inhibitory activity of chiral 5-hydroxy-2-N-acyl-(3E) sphingenines: ceramides with an unusual sphingoid backbone. AB - The unusual sphingoid base 5-hydroxy-3-sphingenine was identified in the hydrolysate of brain sphingolipids more than 40 years ago. We present here the first synthesis of the 5R and 5S diastereoisomers of the N-acyl derivatives of 5 hydroxy-3-sphingenine, 2 and 3, respectively, which represent regioisomers of (2S,3R)-ceramide (1). The key steps include the synthesis of alpha,beta unsaturated ketone intermediates 4 and 5 from N-Cbz- and N-Boc-l-serine and diastereoselective reduction of the enones. The configuration at the new carbinol center was deduced by proton NMR analysis of (R)- and (S)-Mosher [methoxy(trifluoromethyl)phenylacetate] ester derivatives. Ceramide analogues 2 and 3 showed a markedly higher antiproliferative activity than 1 on MCF-7 cells. PMID- 12530862 TI - 6-aza-2'-deoxyisocytidine: synthesis, properties of oligonucleotides, and base pair stability adjustment of DNA with parallel strand orientation. AB - 6-Aza-5-methyl-2'-deoxyisocytidine (1a) and 6-aza-2'-deoxyisocytidine (1b) have been synthesized, converted into phosphoramidite building blocks, and incorporated into oligodeoxynucleotides. The glycosylic bond stability of 1a,b under acidic conditions increases compared to that of 5-methyl-2' deoxyisocytidine (2a) and 2'-deoxyisocytidine (2b). Oligonucleotides incorporating 1a or 1b show an enhanced stability against the 3'-exonuclease snake-venom phosphodiesterase. The duplexes containing 6-azapyrimidine nucleosides 1a or 1b have lower T(m) values than duplexes containing 2a or 2b with either antiparallel or parallel chain orientation. This was used to adjust the stability of the tridentate m5iCd-dG base pair to the level of the bidentate reverse Watson-Crick dA-dT pair. PMID- 12530863 TI - Diastereoselective cobalt-mediated [2 + 2 + 2] cycloadditions of substituted linear enediynes phosphine oxides: scope and limitations. AB - Variously substituted linear enediynes phosphines oxides possessing the double bond at either the terminal or internal position and with the phosphine oxide appended onto the alkyne or the alkene terminus have been prepared. Their cobalt(I)-mediated cyclizations produce the eta(4)-complexed tricyclic compounds in high yields. The endo/exo selectivity depends on both the position of the phosphine oxide on the enediyne and the position of the double bond in the tether. With chiral phosphine oxides, a certain degree of induction was observed, and depending on the substituents on the phosphorus atom, the diastereoselectivity can reach 74%. Up to now, it is the highest level reported for such a cyclization in which a stereogenic center is created. Regarding all of our results, two reaction pathways involving an initial coordination of the cobalt moiety on the chelating site of the substituent have been suggested to explain the observed selectivities. PMID- 12530864 TI - Synthesis of 2-substituted benzofurans and indoles using functionalized titanium benzylidene reagents on solid phase. AB - Titanium(IV) benzylidenes bearing a masked oxygen or nitrogen nucleophile in the ortho position were generated from thioacetals, using low-valent titanocene complex, Cp2Ti[P(OEt)3]2. Methylene acetal, alkyl ether, silyl ether, fluoro, tertiary amino, and N-alkyl, N-benzyl, N-prenyl, and N-silyl tert-butyl carbamate groups were tolerated in the titanium alkylidene reagents (Schrock carbenes). Aryl-chlorine bonds were stable to the titanium benzylidene functionality, but there was poor chemoselectivity for the reduction of the thioacetal in the presence of an aryl chloride. The titanium benzylidenes converted Merrifield and Wang resin-bound esters into enol ethers. The oxygen nucleophile was masked as a TMS ether, and when the resin-bound enol ethers bearing this ortho substituent were treated with 1% TFA in dichloromethane, benzofurans were released from resin in high yields. The chameleon catch strategy ensured excellent purity. In a similar way, N-alkylated and N-silylated tert-butyl carbamates were used for the synthesis of N-alkyl and N-Boc indoles, respectively. These traceless solid-phase syntheses of heterocycles are believed to involve postcleavage modification rather than cyclative termination. PMID- 12530865 TI - Biocatalytic asymmetric hydrogen transfer employing Rhodococcus ruber DSM 44541. AB - Nonracemic sec-alcohols of opposite absolute configuration were obtained either by asymmetric reduction of the corresponding ketone using 2-propanol as hydrogen donor or by enantioselective oxidation through kinetic resolution of the rac alcohol using acetone as hydrogen acceptor employing whole lyophilized cells of Rhodococcus ruber DSM 44541. The microbial oxidation/reduction system exhibits not only excellent stereo- and enantioselectivity but also a broad substrate spectrum. Due to the exceptional tolerance of the biocatalyst toward elevated concentrations of organic materials (solvents, substrates and cosubstrates), the process is highly efficient. The simple preparation of the biocatalyst and its ease of handling turns this system into a versatile tool for organic synthesis. PMID- 12530866 TI - Lithiation of 1-chloromethylbenzotriazole: generation and elaboration of benzotriazolyloxiranes. AB - 1-Benzotriazolylchloromethyllithium generated from 1-chloromethylbenzotriazole (1) and LDA reacts with enolizable and nonenolizable ketones to give benzotriazolyloxiranes 2a-g in good yields. The oxiranyllithiums 4a-d generated from 2a-d and n-BuLi at -78 degrees C were trapped by a variety of electrophiles to give oxiranyl derivatives 5a-j in good to excellent yields. Lewis-acid promoted nucleophilic ring opening of benzotriazolyloxiranes 2a,f,g with allyltrimethylsilane gave the corresponding 1,7-octadien-4-ols 6a-c in 68-75% yield. Hydrolysis of alpha-acylbenzotriazolyloxiranes 5g,h provided 3-hydroxy-1,2 diones 7a and 7b in 73 and 86% yield, respectively. PMID- 12530867 TI - A theoretical study of the mechanisms and regiochemistry of the reactions of 5 alkoxyoxazole with thioaldehydes, nitroso compounds, and aldehydes. AB - A theoretical study based on B3LYP/6-31G calculations has been applied to the mechanisms and regiochemistry of reactions of 5-alkoxyoxazole with thioaldehydes, nitroso compounds, and aldehydes. All three reactions adopt similar mechanisms, which start with Diels-Alder (DA) reactions, followed by either a novel, concerted ring-opening-ring-closing (RORC) step to transfer the DA adduct to 2 alkoxycarbonyl-3-thiazoline and 2-alkoxycarbonyl-3-oxazoline for thioaldehydes and aldehydes, respectively, or stepwise ring-opening and ring-closing steps to generate 1,2,4-oxadiazoline for nitroso compounds. The reactions of 5 alkoxyoxazole with thioaldehydes and nitroso compounds can be conducted under thermal reaction conditions due to the 10 kcal/mol activation barriers for their rate-determining DA reactions. By contrast, the reaction of 5-alkoxyoxazole with aldehydes cannot take place under thermal conditions, since this bimolecular reaction has the rate-determining RORC transition state higher than the reactants by 30.5 kcal/mol. PMID- 12530868 TI - A DFT study of the mechanisms and regio- and stereochemistry of the lewis acid catalyzed reactions of 5-alkoxyoxazoles with aldehydes: aryl substitution at the 2-position of 5-alkoxyoxazole is critical to the formation of 4-alkoxycarbonyl-2 oxazoline. AB - Density functional theory at the B3LYP/6-31G level has been used to study the mechanisms and regio- and stereochemistry of the Lewis acid-catalyzed reactions of aldehydes with 5-alkoxyoxazoles. Similar to the uncatalyzed reaction between aldehyde and 5-methoxyoxazole, which has an activation energy of 30.5 kcal/mol and intrinsically favors production of 2-alkoxycarbonyl-3-oxazoline, the Lewis acid-catalyzed reaction also prefers to generate 2-alkoxycarbonyl-3-oxazoline in a more efficient way with an activation energy of about 3 kcal/mol (with respect to separated acetaldehyde-AlCl3 complex and 5-methoxyoxazole) in the gas phase. Only when an aryl group is introduced to the 2 position of 5-alkoxyoxazoles can the Lewis acid-catalyzed reactions furnish 4-alkoxycarbonyl-2-oxazolines. The reasons for this switch of regiochemistry and the factors affecting the stereochemistry are discussed. PMID- 12530869 TI - Photochemical, thermal, and base-induced access to hydroazulene derivatives via two-carbon ring-enlargement reactions of condensed electrophilic cyclobutenes. AB - Enamines, silyl enol ethers, and beta-keto ester anions derived from bicyclo[3.3.0]octan-2-one efficiently underwent a formal [2 + 2] cycloaddition reaction with DMAD and ethyl propynoate leading to a large variety of electrophilic cyclobutenes. The latter were transformed into polyfunctionalized bicyclo[5.3.0]decane (or hydroazulene) ring systems in high yields by fragmentation of the cyclobutene moiety. These two-carbon ring-enlargement reactions were utilized as a synthetic tool for the construction of a polyfunctionalized hydroazulene derivative that represents a potential precursor of the tricyclic framework of ingenol. PMID- 12530871 TI - P(i-BuNCH2CH2)3N: an effective ligand in the palladium-catalyzed amination of aryl bromides and iodides. AB - It is shown that the bicyclic triaminophosphine P(i-BuNCH2CH2)3N serves as an effective ligand for the palladium-catalyzed amination of a wide array of aryl bromides and iodides. Other bicyclic or acyclic triaminophosphines, even those of similar basicity and/or bulk, were inferior. PMID- 12530870 TI - Aromatic nucleophilic substitution or CuI-catalyzed coupling route to martinellic acid. AB - Condensation of beta-amino ester 8b with triflate 7 gives N-aryl amino ester 11, which is converted into 2-substituted 4-oxoquinoline 4 using an intramolecular Dieckmann reaction as the key step. CuI-mediated coupling of beta-amino ester 8a with 1,4-diiodobenzene followed by an intramolecular acylation and Pd-catalyzed carbonylation provide another manner to 4. Alkylation of 4 and subsequent reductive amination deliver the cyclic imine 14, which is transformed into triamine 3 by ordinary operations. Guanylation of 3 under mild condition followed by deprotection results in the synthesis of martinellic acid 1. PMID- 12530872 TI - Synthesis of oxazines and N-arylpyrroles by reaction of unfunctionalized dienes with nitroarenes and carbon monoxide, catalyzed by palladium-phenanthroline complexes. AB - The reaction between an unfunctionalized conjugated diene and a nitroarene under CO pressure and at 100 degrees C, catalyzed by [Pd(Phen)2][BF4]2 (Phen = 1,10 phenanthroline), affords the corresponding hetero-Diels-Alder adduct (oxazine) in up to 91% yields in one pot. If the reaction mixture is then heated to 200 degrees C, the oxazines are converted into the corresponding N-arylpyrroles in good yields. Pressures as low as 5 bar can be employed, and 0.08% catalyst is sufficient to effect the transformation. The reaction can be equally run by employing the nitroarene or the diene as limiting agent and works well for nitroarenes bearing either electron-withdrawing or mildly electron-donating substituents. A moderate steric hindrance on the nitroarene (o-methyl) is well tolerated, but 1,4-disubstituted-1,3-dienes are not suitable substrates. PMID- 12530873 TI - Highly regioselective Friedlander annulations with unmodified ketones employing novel amine catalysts: syntheses of 2-substituted quinolines, 1,8-naphthyridines, and related heterocycles. AB - Catalysts were evaluated on the preparation of 2-substituted quinolines, 1,8 naphthyridines, and chromone derivatives from unmodified methyl ketones and o aminoaromatic aldehydes. While oxide catalysts yielded the 2,3-dialkyl substituted products, cyclic secondary amines provided the 2-alkylsubstutited products regioselectively. In particular, pyrrolidine derivatives provided the highest regioselectivity favoring the 2-substituted products. The most reactive and regioselective catalyst was the bicyclic pyrrolidine derivative, TABO (1,3,3 trimethyl-6-azabicyclo[3.2.1]octane), yielding 1,8-naphthyridines with as high as 96:4 regioselectivity. Regioselectivity increased with slow addition of the methyl ketone substrate to the reaction mixture, and was positively related to temperature. Isolated yields of single regioisomers were typically 65-84%, while observed regioselectivities were > or =90:10 for 1,8-naphthyridines and > or =84:16 for quinolines. PMID- 12530875 TI - The aminomethylation of electron-rich aromatics with an N-silyl-N,O-acetal catalyzed by a metal triflate-TMSCl system: facile synthesis of aromatic primary amines, 1-aryl-trichloroethylamines. AB - The copper(II) triflate- and hafnium(IV) triflate-catalyzed aminomethylation of indole (2) with an N-silyl-N,O-acetal 1 containing a trichloromethyl group provides the primary amine derivative (3a) in modest yield. When 1 equiv of trimethylchlorosilane (TMSCl) was added to the reaction mixture, the reaction proceeded smoothly, and the yield of 3a was dramatically improved (>90%). The use of this catalytic system permitted the introduction of an aminomethyl group onto indoles 2a-h bearing a variety of functional groups, which appears to deactivate the Lewis acid, in 52-92% yields. Hf(OTf)4-doped TMSCl catalyzed the successful aminomethylation of various electron-rich aromatic compounds 4a-j to produce 1 aryl-trichloroethylamine derivatives 5a-j. PMID- 12530874 TI - Formation of a methide derivative upon photolysis of thymidine bromohydrins. AB - Reaction of bromine with thymidine in aqueous solution produces, in high yield, the corresponding 5-bromo-6-hydroxy-5,6-dihydroderivative (thymidine bromohydrins). UVC photolysis of thymidine bromohydrins gives rise to a reactive intermediate that is converted into 5-(hydroxymethyl)-2'-deoxyuridine upon incubation in water. When the former compound is left in methanol, ethanol, or propanol, the corresponding 5-alkoxymethyl derivatives are produced. The proposed structure for the primary photolysis product of thymidine bromohydrins is a methide derivative of the thymine ring. This compound could be an interesting intermediate in the synthesis of methyl-substituted thymidine. PMID- 12530876 TI - Carbodiimide-mediated preparation of the tricyclic pyrido[3',2':4,5]pyrrolo[1,2 c]pyrimidine ring system and its application to the synthesis of the potent antitumoral marine alkaloid variolin B and analog. AB - A total synthesis of the marine alkaloid variolin B has been completed in 13 steps in an overall yield of 6.5% from 3-formyl-4-methoxypyridine. Our approach is based on the sequential formation of the 7-azaindole ring, the tricyclic pyrido[3',2':4,5]pyrrolo[1,2-c]pyrimidine ring system, and finally installation of the 2-aminopyrimidine ring at C5. The required 7-azaindole ring appropriately substituted is formed by a modified indole synthesis involving a nitrene insertion process (two steps). Formation of the annelated pyrimidine ring is achieved by two routes both involving a carbodiimide-mediated cyclization process, which allow incorporation of the amine functionality at C9 of the core tricyclic (six steps). Installation of the northeast 2-aminopyrimidine ring at C5 is performed using the Bredereck protocol (three steps). Ultimate, thermal decarboxylation with concomitant O-methyl deprotection and further N-benzyl deprotection by the action of triflic acid completed the synthesis of the target natural product variolin B. PMID- 12530877 TI - Unexpected differences in the alpha-halogenation and related reactivity of sulfones with perhaloalkanes in KOH-t-BuOH. AB - Most alkyl phenyl sulfones are readily alpha-chlorinated with CCl(4) and alpha brominated with CBrCl3 in KOH-t-BuOH via radical-anion radical pair (RARP) reactions. While isopropyl mesityl sulfone (4) is easily alpha-chlorinated with CCl(4), it was completely recovered when treated with the more reactive CBrCl3. Subsequent investigations showed the latter result to be due to the poor acidity of 4 together with the rapid depletion of CBrCl3 and KOH by their reaction with each other, and led to a variety of other important results. 4-Hydroxyphenyl isopropyl sulfone (6) is unreactive with either CCl4 or CBrCl3 in KOH-t-BuOH, its phenoxide anion strongly reducing the electronegativity of the sulfonyl group, thereby inhibiting alpha-anion formation. This effect is reversed by the electron withdrawing influence of two alpha-phenyls, so that benzhydryl 4-hydroxyphenyl sulfone (8) is readily alpha-halogenated in KOH-t-BuOH with CCl4 or CBrCl3. On further contact with KOH-t-BuOH the alpha-halogenated sulfones from 8 are decomposed into benzophenone and phenol. While the alpha-halogenated derivatives of 4-methoxyphenyl benzhydryl sulfone (9) are stable to base, they are decomposed even under mildly acidic conditions into 4-methoxyphenyl 4 methoxybenzenethiolsulfonate (9c), phenol, and benzophenone. Mono-alpha halogenation of benzyl phenyl sulfone (10) enhances the rate of the subsequent halogenation, so that alpha,alpha-dihalogenation is attained while much substrate is still present and the mono-alpha-halogenated product is not detected. The ease of reductive debromination of alpha-bromo sulfones with Cl3C- was correlated with the stability of the formed alpha-anions, explaining the success with alpha bromobenzylic sulfones but failure with alpha-bromoalkyl sulfones. In the presence of air and the absence of competing halogenation, formation of the alpha anions of alkyl aryl sulfones is quickly accompanied by oxidative cleavage by atmospheric O2, leading to the formation of arenesulfonyl alcohols, arenesulfonyl halides, and haloarenes. PMID- 12530878 TI - Versatile routes to C-2- and C-6-functionalized glucose derivatives of iminodiacetic acid. AB - A series of novel d-glucose derivatives, functionalized at the C-2 or the C-6 position with an iminodiacetic acid moiety for transition-metal complexation, has been prepared. The sugar and the metal-chelating parts are separated by either propyl or octyl chains and were introduced by the reaction of bromoalkylamine. Either N-1-Boc-3-bromopropylamine (17) or N-(8-bromooctyl)phthalimide (19) reacted with methyl 3,5,6-tri-O-benzyl-alpha-beta-d-glucofuranoside (4) (C-2 position) and 1,2:3,5-(O-methylene)-alpha-d-glucose (11) (C-6 position), respectively, in the presence of sodium hydride in DMF at room temperature, affording the desired intermediates. For aminopropyl derivatives, yields varied between 57% and 65%, and for aminooctyl derivatives, yields varied between 40% and 71%. After deprotection of the amine functionality, the metal chelate was built up by dialkylation (6a-c and 13a,b) with methyl bromoacetate in the presence of triethylamine under reflux in THF. Yields varied between 56% and 69% for the glucose modified at the C-2 position and between 58% and 62% for the one modified at the C-6 position. All compounds were characterized by 1H or 13C NMR or both, IR, and mass spectroscopy. Final products were isolated as a mixture of alpha and beta anomers. PMID- 12530879 TI - A correlation between the absolute configuration of alkyl aryl sulfoxides and their helical twisting powers in nematic liquid crystals. AB - In this paper, for the first time, a systematic experimental and theoretical analysis of the cholesteric induction due to solutes whose chirality is originated only by a single stereogenic center has been carried out. The twisting power beta of a series of alkyl aryl sulfoxides has been determined in several nematic solvents. The sign of beta, which reflects the handedness of the induced helical arrangement of the solvent molecules, correlates with the configuration of the stereogenic sulfur in the nematic solvents E7, Phase 1083, and ZLI 2359: (S)-configurated dopants induce (M)-chiral nematics. (S)-Configurated cyclic sulfoxides, which are forced to adopt a different conformation with respect to the parent acyclic compounds, induce, instead, right-handed chiral nematics. The experimental data have been interpreted in the light of the surface chirality model, which allows the calculation of beta in terms of the molecular properties of the dopant, namely, the anisotropy and helicity of its molecular surface. The calculations reliably reproduce the behavior experimentally observed. The more flexible, open-chain compounds investigated induce chiral nematics of opposite handedness in MBBA and Phase 1053: temperature-dependent experiments point out the importance of the conformation in determining the effective sign of beta. The results have been discussed in terms of different conformation populations in these latter solvents with respect to E7, Phase 1083, and ZLI 2359. PMID- 12530880 TI - Synthesis of novel bis(beta-cyclodextrin)s and metallobridged bis(beta cyclodextrin)s with 2,2'-diselenobis(benzoyl) tethers and their molecular multiple recognition with model substrates. AB - To investigate quantitatively the cooperative binding ability of beta cyclodextrin dimers, a series of bridged bis(beta-cyclodextrin)s with 2,2' diselenobis(benzoyl) spacer connected by different lengths of oligo(ethylenediamine)s (2-5) and their platinum(IV) complexes (6-9) have been synthesized and their inclusion complexation behavior with selected substrates, such as Acridine Red, Neutral Red, Brilliant Green, Rhodamine B, ammonium 8 anilino-1-naphthalenesulfonate, and 6-p-toluidino-2-naphthalenesulfonic acid, were investigated by means of ultraviolet, fluorescence, fluorescence lifetime, circular dichroism, and 2D-NMR spectroscopy. The spectral titrations have been performed in aqueous phosphate buffer solution (pH 7.20) at 25 degrees C to give the complex stability constants (K(S)) and Gibbs free energy changes (-DeltaG degrees ) for the inclusion complexation of hosts 2-9 with organic dyes and other thermodynamic parameters (DeltaH degrees and TDeltaS degrees ) for the inclusion complexation of 2-5with fluorescent dyes ANS and TNS. The results obtained indicate that beta-cyclodextrin dimers 2-5 can coordinate with one or two platinum(IV) ions to form 1:1 or 1:2 stoichiometry metallobridged bis(beta cyclodextrin)s. As compared with parent beta-cyclodextrin (1) and bis(beta cyclodextrin)s 2-5, metallobridged bis(beta-cyclodextrin)s 6-9 can further switch the original molecular binding ability through the coordinating metal to orientate two beta-cyclodextrin cavities and an additional binding site upon the inclusion complexation with model substrates, giving the enhanced binding constants K(S) for both ANS and TNS. The tether length between two cyclodextrin units plays a crucial role in the molecular recognition with guest dyes. The binding constants for TNS decrease linearly with an increase in the tether length of dimeric beta-cyclodextrins. The Gibbs free energy change (-DeltaG degrees ) for the unit increment per ethylene is 0.32 kJ.mol(-)(1) for TNS. Thermodynamically, the higher complex stabilities of both ANS and TNS upon the inclusion complexation with 2-5 are mainly contributed to the favorable enthalpic gain (-DeltaH degrees ) by the cooperative binding of one guest molecule in the closely located two beta-cyclodextrin cavities as compared with parent beta cyclodextrin. The molecular binding ability and selectivity of organic dyes by hosts 1-9 are discussed from the viewpoints of the multiple recognition mechanism and the size/shape-fitting relationship between host and guest. PMID- 12530881 TI - Regioselective synthesis of substituted o-alkoxyphenol derivatives through thermal benzannulation of Fischer (alkenylcyclobutenyl)carbene complexes. AB - A convenient, regioselective, and general synthetic method for producing highly substituted o-phenol-containing polycycles from Fischer (alkenylcyclobutenyl)carbene complexes has been described. The starting complexes have been synthesized by means of the [2 + 2] cycloaddition reaction of (alkenylethynyl)carbene complexes and a range of enol ethers, and in most cases, they have proven to be stable at room temperature and therefore isolable. The key step of the synthesis consists of the thermal benzannulation reaction of these novel pentacarbonyl dienyl Fischer complexes, which is an unprecedented transformation for these kinds of complexes. The unexpected behavior of (alkenylcyclobutenyl)carbene complexes has been rationalized in terms of their geometries. PMID- 12530882 TI - Total synthesis of (-)-fumiquinazolines A, B, C, E, H, and I. Approaches to the synthesis of fiscalin A. AB - The first syntheses of (-)-fumiquinazolines A, B, and I, which proceed in 14 steps from protected tryptophan, anthranilic acid, leucine, and alanine in 7% overall yield, are described. Tricycle 30 was formed by a palladium-catalyzed cyclization. Oxidation of 30a with saccharine-derived oxaziridine 21 for fumiquinazolines A and B and oxidation of 30b with dimethyldioxirane for fumiquinazoline I selectively formed the appropriate imidazoindolone stereoisomers. Application of the Ganesan-Mazurkiewicz cyclization completed the syntheses. Efficient 14-step syntheses of (-)-fumiquinazolines C (7) and E (3) and a 15-step synthesis of (-)-fumiquinazoline H (8) using FmocNHCH(CH2SePh)CO2H as a dehydroalanine precursor that spontaneously eliminated benzeneselenol without oxidation under the cyclization conditions are also reported. Model 86 for fiscalins A with the H and OH anti to each other has been prepared, but the procedure that worked for the model failed with the fully functionalized side chain. PMID- 12530883 TI - Theoretical and experimental studies of asymmetric organozinc additions to benzaldehyde catalyzed by flexible and constrained gamma-amino alcohols. AB - The cis-decalin based gamma-amino alcohols, 1-5, were synthesized, resolved, and employed as catalysts in the addition of organozincs to benzaldehyde. Despite large facial differentiation in the RZn adducts of the ligands, the enantioselectivities were found to be moderate. Transition structure calculations were done using several quantum chemical methods to examine the underlying causes of the selectivity for three gamma-amino alcohols, 1, 2, and 3. The tricyclic mu oxo (6/4/4), rather than the bicyclic six-membered (6/6), transition structures were found to explain the observed enantioselectivity for 2 and 3. MP2 calculations gave the best correlation to the experimental results compared to B3LYP and HF calculations. The conformational flexibility of the chiral ligands was found to be an important factor in the selectivity. PMID- 12530885 TI - Generation of aryl radicals from arylboronic acids by manganese(III) acetate: synthesis of biaryls and heterobiaryls. AB - The efficient generation of aryl radicals from arylboronic acids by manganese(III) acetate is described. In aromatic solvents, in situ generated aryl radicals afford the corresponding biaryls in very good yields. This method works selectively, and yields are better than those from similar, previously described methods. Arylboronic acids carrying sensitive functional groups also work efficiently. PMID- 12530884 TI - Origins of boat or chair preferences in the Ireland-Claisen rearrangements of cyclohexenyl esters: a theoretical study. AB - The relative energies of the chair and boat transition states of a variety of Ireland-Claisen rearrangements were obtained by B3LYP/6-31G calculations. Theoretical results are in good agreement with experimental data and provide a quantitative analysis of the origins of boat preferences that are observed in some of these reactions. PMID- 12530886 TI - Catalytic enantioselective [3 + 2]-cycloadditions of diazoketone-derived aryl substituted carbonyl ylides. AB - An evaluation of alpha-aryl-alpha-diazodiones in tandem carbonyl ylide formation enantioselective [3 + 2]-cycloaddition reactions is described. Such substrates were designed to allow investigation of the electronic characteristics of the dipole upon asymmetric induction. Intramolecular cycloadditions (with a tethered alkene dipolarophile) were found to occur in good to quantitative yields, with a difference in ee exhibited by the two electronically different diazodiones 8 and 9. Intermolecular cycloadditions using diazodiones 12 and 13 with DMAD and arylacetylenes 16-18 again demonstrated that electronics play a key role in determining the outcome of the cycloaddition reactions. Enantioselectivities of up to 76% were observed. PMID- 12530887 TI - Green chemistry approaches to the synthesis of 5-alkoxycarbonyl-4-aryl-3,4- dihydropyrimidin-2(1H)-ones by a three-component coupling of one-pot condensation reaction: comparison of ethanol, water, and solvent-free conditions. AB - A general and practical green chemistry route to the Biginelli cyclocondensation reaction using cerium(III) chloride as the catalyst (25% mol) is described under three different sets of reaction conditions. This method provides an efficient and much improved modification of original Biginelli reaction reported in 1893, in terms of high yields, short reaction times, and simple work-up procedure, and it has the ability to tolerate a wide variety of substitutions in all three components, which is lacking in existing procedures. PMID- 12530888 TI - Synthesis of chiral ionic liquids from natural amino acids. AB - For the first time, chiral imidazolium ionic liquids containing one chiral carbon (10a-c) were synthesized from the natural amino acids by a simple and straightforward procedure. The characteristics of the chiral ILs are very similar to the popular ionic liquids. PMID- 12530889 TI - Diastereoselectivity in the reduction of alpha-Oxy- and alpha-amino-substituted acyclic ketones by polymethylhydrosiloxane. AB - Diastereoselectivity in the reduction of alpha-alkoxy-, alpha-acyloxy-, and alpha alkylamino-substituted ketones with polymethylhydrosiloxane (PMHS) in the presence of fluoride ion catalysis was investigated. High syn-selectivity was observed in the reduction of alpha-alkoxy, alpha-acyloxy, and alpha-dialkylamino ketones. Reduction of alpha-monoalkylamino ketone proceeded in anti-selective manner with moderate selectivity. The observed selectivity is explained based on Felkin-Anh and Cram-chelate models. PMID- 12530890 TI - Molecular taekwondo. 2. A new calix[4]azacrown bearing two different binding sites as a new fluorescent ionophore. AB - Synthesis and binding studies of a new calixarene-based fluoroionophore were made. Calix[4]azacrown having an anthracenyl unit displayed large chelation enhanced fluorescence effects with Cs(+), Rb(+), and K(+) over other metal ion tested. Interesting "molecular taekwondo" processes between Cs(+)-Cu(2+) and Cs(+)-Ag(+) pairs were monitored via fluorescent changes. PMID- 12530891 TI - An improved procedure for the palladium-catalyzed oxidative carbonylation of beta amino alcohols to oxazolidin-2-ones. AB - A highly efficient oxidative cyclocarbonylation of beta-amino alcohols and 2 aminophenol to oxazolidin-2-ones has been achieved by using PdI(2) in conjunction with KI as the catalytic system in DME under relatively mild conditions (100 degrees C and 20 atm of a 4:1 mixture of CO and air). PMID- 12530892 TI - Fluorinated heterocyclic compounds. An expedient route to 5-perfluoroalkyl-1,2,4 triazoles via an unusual hydrazinolysis of 5-perfluoroalkyl-1,2,4-oxadiazoles: first examples of an ANRORC-like reaction in 1,2,4-oxadiazole derivatives. AB - The hydrazinolysis reaction of 5-perfluoroalkyl-1,2,4-oxadiazoles has been investigated. Nucleophilic addition of the reagent to the C(5)-N(4) double bond of the oxadiazole ring, followed by ring-opening and then ring-closure involving the beta-nitrogen atom of the hydrazino moiety and the C(3) of the oxadiazole ring, explains the formation of 5-perfluoroalkyl-1,2,4-triazoles as final products. Useful applications in synthesis of this uncommon hydrazinolysis can be claimed. PMID- 12530893 TI - Click chemistry to construct fluorescent oligonucleotides for DNA sequencing. AB - "Click chemistry" 1,3-dipolar cycloaddition between alkynyl 6-carboxyfluorescein (FAM) and azido-labeled single-stranded (ss) DNA was carried out under aqueous conditions to produce FAM-labeled ssDNA in quantitative yield. The FAM-labeled ssDNA was successfully used as a primer to produce DNA sequencing products with single-base resolution in a capillary electrophoresis DNA sequencer with laser induced fluorescence detection. PMID- 12530894 TI - Selective cleavage of N-benzyl-protected secondary amines by triphosgene. AB - A series of competition experiments has revealed that selective cleavage of N benzyl-protected secondary amines can be achieved with triphosgene, thereby providing a useful range of carbamoyl chlorides. PMID- 12530895 TI - An expedient approach to internally functionalized chiral dendrimers: synthesis of a dendritic molecule incorporating furanoside skeleton. AB - In an approach to chiral dendritic molecules, a dendrimer incorporating pentose units in the interior and hexose units in the periphery is built up on a 1, 3, 5 trisubstituted aromatic core by using 1,2:5,6-diisopropylidene glucose as the carbohydrate precursor and a 3, 5-disubstituted aromatic unit as the branching block. The carbohydrate moiety also provides internal functionalities in the form of hemiacetal moiety of the furanoside ring. PMID- 12530897 TI - Hydroxyalkylation of alpha-C-H bonds of tetrahydrofuran with aldehydes in the presence of triethylborane and tert-butyl hydroperoxide. AB - The alpha-hydroxyalkylation of tetrahydrofuran with aldehydes via radical C-H abstraction was conducted using triethylborane in the presence of tert-butyl hydroperoxide. This study presents a rare instance of direct intermolecular radical addition of unactivated cyclic ether to aldehydes. PMID- 12530896 TI - Nitrile biotransformation for highly enantioselective synthesis of 3-substituted 2,2-dimethylcyclopropanecarboxylic acids and amides. AB - Biotransformations of differently configured 2,2-dimethyl-3-substitued cyclopropanecarbonitriles were studied using a nitrile hydratase/amidase containing Rhodococcus sp. AJ270 whole-cell catalyst under very mild conditions. Although all of the cis-3-aryl-2,2-dimethylcyclopropanecarbonitriles appeared inert toward the biocatalyst, a number of racemic trans-isomers efficiently underwent a highly enantioselective hydrolysis to produce (+)-(1R,3R)-3-aryl-2,2 dimethylcyclopropanecarboxylic acids and (-)-(1S,3S)-3-aryl-2,2 dimethylcyclopropanecarboxamides in high yields with excellent enantiomeric excesses in most cases. The overall enantioselectivity of the biotransformations of nitriles originated from the combined effects of 1R-enantioselective nitrile hydratase and amidase, with the later being a dominant factor. The influence of the substrates on both reaction efficiency and enantioselectivity was discussed in terms of steric and electronic effects. Coupled with chemical transformations, biotransformations of nitriles provided convenient syntheses of optically pure geminally dimethyl-substituted cyclopropanecarboxylic acids and amides, including chrysanthemic acids, in both enantiomeric forms. PMID- 12530898 TI - Improved method for the synthesis of 2-alkylamino-2-deoxy-d-glucopyranose and 1,2 dialkylamino-1,2-dideoxy-d-(N)-beta-glucoside. AB - An improved method for the synthesis of two types of carbohydrate-based, pH sensitive surfactants has been developed. The surfactants are formed in almost quantitative yields from d-fructose and suitable alkylamines, when using a zinc halide salt as catalyst. The reaction proceeds via an initial Amadori reaction, and the 1,2-dialkylamino-1,2-dideoxy-d-(N)-beta-glucoside precipitates out of the reaction. The product can easily be hydrolyzed to the more water-soluble, surface active, and pH-sensitive monoalkyl derivative 2-alkylamino-2-deoxy-d glucopyranose. Products from several amines can be prepared. PMID- 12530899 TI - Electrochemical synthesis of alkyl nitroaromatic compounds. AB - Alkyl nitroaromatic compounds were readily prepared via nucleophilic aromatic substitution for hydrogen or a heteroatom by electrochemical oxidation of the sigma-complex. Butyllithium and butylmagnesium chloride were used as nucleophiles, and several nitrocompounds were tested to explore the possibilities of the NASH and NASX reactions promoted electrochemically. PMID- 12530900 TI - Photoactivated isomerization of linear olefins. AB - The photocatalyzed (350 nm) isomerization of linear olefins catalyzed by platinum(II) bis(acetylacetonato) [Pt(acac)(2)] in the presence of silanes was studied. Catalytic activity depends on the silane. Triphenylsilane is the most reactive silane studied, with more than 98% isomerization of the alkene occurring after 20-min irradiation. There is no concomitant hydrosilylation. The mechanism has been investigated and a metal hydride addition-elimination mechanism proposed. PMID- 12530901 TI - Crystal structure and rotational barrier of octakis(bromomethyl)naphthalene. AB - Octakis(bromomethyl)naphthalene (4) adopts in the crystal a chiral conformation with a helical central naphthalene core and the bromomethyl groups disposed in an alternate up-down "in" arrangement. According to MM3 calculations, this conformation is less stable than the corresponding all alternated "out" form, while B3LYP/LANL2DZ calculations suggest the opposite stability order. The topomerization barrier (16.0 kcal mol(-1)) is ascribed to an enantiomerization process requiring 180 degrees rotation of all the bromomethyl groups and reversal of the helical sense of the naphthalene core. PMID- 12530902 TI - Kinetic enolate formation by lithium arylamide: effects of basicity on selectivity. AB - Five ketones R1COCH2R2 (1a-e) were enolized in tetrahydrofuran solvent employing lithium arylamides with different electron-withdrawing and -donating substituents on the phenyl ring (4a-e). Enolate selectivity is unaffected by a moderate electron-releasing or -withdrawing group, but significantly enhanced by strong electron-withdrawing substituents to yield predominantly Z-enolate. Outstanding selectivity was achieved with lithium trichloroanilide (5) and lithium diphenylamide (6). The results are rationalized in terms of electronic effects on the tightness of the transition states. PMID- 12530903 TI - An efficient assembly of heterobenzazepine ring systems utilizing an intramolecular palladium-catalyzed cycloamination. AB - Azaheterocyclic compounds are interesting and medicinally relevant targets. Herein we disclose an improved synthesis into the oxazepine and thiazepine ring systems. The key step in the synthesis exploits recent advancements in the palladium-catalyzed amination reaction, which was utilized to form the seven membered rings. General conditions for this reaction were Pd2dba3, P(t-Bu)3, NaO t-Bu alone or with K2CO3, in toluene. The scope of the reaction was investigated, and has been shown to be effective on a variety of substrates as illustrated. PMID- 12530904 TI - Synthesis of (R)-3,4-diaminobutanoic acid by desymmetrization of dimethyl 3 (benzylamino)glutarate through enzymatic ammonolysis. AB - Lipase B from Candida antarctica is shown to be a highly efficient catalyst for the desymmetrization of dimethyl 3-(benzylamino)glutarate (1) through aminolysis and ammonolysis reactions. Using this procedure, enantiopure monoamides are obtained in high yield. The synthetic value of these compounds is demonstrated by the preparation of an enantiopure nonnatural amino acid, i.e. (R)-3,4 diaminobutanoic acid [(+)-12]. PMID- 12530905 TI - [5]helicene-fused phthalocyanine derivatives. New members of the phthalocyanine family. AB - An efficient synthetic route to fuse [5]helicene moieties around the phthalocyanine core is reported. The helicene moiety was constructed by the Diels Alder reaction of 3,4,3',4'-tetrahydro-1,1'-dinaphthyl and dibromobenzyne. Subsequent cyanation, oxidation, O-alkylation, and cyclic tetramerization resulted in octaalkoxy phthalocyanine derivatives which showed high solubility in common organic solvents and displayed strong absorption in the near-IR region. PMID- 12530906 TI - Preparation and reactivity of O2-sulfonated diazeniumdiolates. AB - We report the facile preparation of O2-sulfonated diazeniumdiolates and mechanistic investigation of their reactions with representative nucleophiles. This new class of compounds extends the range of O2-substituted diazeniumdiolates available for potential applications in research and medicine. PMID- 12530907 TI - Selective synthesis of either enantiomer of alpha-amino acids by switching the regiochemistry of the tricyclic iminolactones prepared from a single chiral source. AB - Preparation of l-alpha-amino acids was easily accomplished simply by exchanging the position of the lactone group of our recently reported chiral template 1 from C2 to C3. The new chiral template 7 was prepared in 54% overall yield over five steps from (1R)-(+)-camphor. Alkylation of iminolactone 7 afforded the alpha monosubstituted products in good yields and excellent diastereoselectivities (>98%). Hydrolysis of the alkylated iminolactones furnished the desired l-alpha amino acids in good yields and ee with nearly quantitative recovery of chiral auxiliary 4. PMID- 12530908 TI - Stereoselective synthesis of enynes by nickel-catalyzed cross-coupling of divinylic chalcogenides with alkynes. AB - (Z,Z)- and (E,E)-divinylic selenides and telurides undergo direct coupling with terminal alkynes in the presence of a nickel/CuI catalyst at room temperature to give (Z)- and (E)-enyne systems in good yields and with complete retention of configuration. PMID- 12530909 TI - Nucleic acid related compounds. 118. Nonaqueous diazotization of aminopurine derivatives. Convenient access to 6-halo- and 2,6-dihalopurine nucleosides and 2' deoxynucleosides with acyl or silyl halides. AB - Treatment of 9-(2,3,5-tri-O-acetyl-beta-d-ribofuranosyl)-2-amino-6-chloropurine (1) with TMS-Cl and benzyltriethylammonium nitrite (BTEA-NO2) in dichloromethane gave the crystalline 2,6-dichloropurine nucleoside 2, and acetyl chloride/BTEA NO2 was equally effective ( approximately 85%, without chromatography). TMS Br/tert-butyl nitrite/dibromomethane gave crystalline 2-bromo-6-chloro analogue 3 (85%). (Chloro or bromo)-dediazoniation of 3',5'-di-O-acetyl-2'-deoxyadenosine (4) gave the 6-[chloro (5, 63%) or bromo (6, 80%)]purine deoxynucleosides, and 2',3',5'-tri-O-acetyladenosine (8) was converted into the 6-chloropurine nucleoside 9 (71%). PMID- 12530910 TI - Palladium-catalyzed cross-coupling reactions of 4-tosyl-2(5H)-furanone with boronic acids: a facile and efficient route to generate 4-substituted 2(5H) furanones. AB - An efficient and facile synthesis of 4-substituted 2(5H)-furanones using palladium catalyzed cross-coupling reactions between 4-tosyl-2(5H)-furanone and boronic acids is reported herein. PMID- 12530912 TI - Mechanisms underlying oocyte activation and postovulatory ageing. AB - Mammalian oocytes undergo significant growth during oogenesis and experience extensive cytoplasmic and nuclear modifications immediately before ovulation in a process commonly referred to as oocyte maturation. These changes are intended to maximize the developmental success after fertilization. Entry of a spermatozoon into the oocyte, which occurs a few hours after ovulation, initiates long-lasting oscillations in the free intracellular calcium ([Ca(2+)](i)) that are responsible for all events of oocyte activation and the initiation of the developmental programme that often culminates in the birth of young. Nevertheless, the cellular and molecular changes that occur during maturation to optimize development are transient, and exhibit rapid deterioration. Moreover, fertilization of oocytes after an extended residence in the oviduct (or in culture) initiates a different developmental programme, one that is characterized by fragmentation, programmed cell death, and abnormal development. Inasmuch as [Ca(2+)](i) oscillations can trigger both developmental programmes in mammalian oocytes, this review addresses one of the mechanism(s) possibly used by spermatozoa to initiate these persistent [Ca(2+)](i) responses, and the cellular and molecular changes that may underlie the postovulatory cellular fragmentation of ageing mammalian oocytes. PMID- 12530913 TI - Prolactin family miniarray: a tool for evaluating uteroplacental-trophoblast endocrine cell phenotypes. AB - The prolactin family represents a group of hormones and cytokines that participate in the control of maternal and fetal adaptations to pregnancy. The aim of this study was to develop a simple assay for monitoring patterns of prolactin family gene expression in rats and mice. Prolactin family cDNAs were spotted on to nylon membranes. Total RNA was extracted from tissues or cells. cDNAs were generated, radiolabelled using reverse transcriptase, and used as probes to hybridize with the prolactin family miniarrays. Pituitary, uterine decidual tissue and placenta each expressed a unique profile of prolactin family members. Placental tissues exhibited regional- and temporal-specific patterns of expression. Prolactin family gene expression differed markedly in mid-pregnant versus late gestation placental tissues and between the junctional and labyrinthine zones of the chorioallantoic placenta. Marked changes in prolactin family gene expression were also observed in cultured trophoblast cells undergoing differentiation. In conclusion, the prolactin family miniarray assay is an effective method for evaluating the endocrine phenotype of the uterus, placenta and trophoblast cells. PMID- 12530914 TI - A re-evaluation of sperm protein 17 (Sp17) indicates a regulatory role in an A kinase anchoring protein complex, rather than a unique role in sperm-zona pellucida binding. AB - Sp17 was originally proposed to be a sperm-specific protein and thought to play a role in sperm-egg interactions by binding to the zona pellucida via two conserved heparin-binding motifs. However, more recent data indicate that it may be expressed more widely, both in tumours and in normal somatic tissues. The complete nucleotide and deduced amino acid sequences of human Sp17 transcripts are reported and it is shown that they are expressed in a wide range of tissues, albeit at a much lower expression than in the testis. On the basis of the extremely high sequence conservation throughout the N-terminal half of Sp17, and the presence within this region of an A-kinase anchoring protein (AKAP)-binding motif, it is postulated that the proposed role of Sp17 in zona pellucida binding is unlikely to be its principal function. PMID- 12530915 TI - Effects of oocyte culture density on meiotic competence of canine oocytes. AB - This study was conducted to determine a suitable ratio of oocytes to medium for in vitro maturation (IVM) of cumulus-oocyte complexes (COCs) collected from bitches at anoestrus and dioestrus and to examine the meiotic competence of COCs cultured singly or in different group sizes. In the first experiment, different numbers of COCs (5, 10, 15 and 20 per drop) were cultured for 72 h in 100 microl drops of maturation medium. The meiotic competence of oocytes from ovaries at anoestrus was affected by the number of COCs incubated, whereas at dioestrus, the incubation number of COCs had no effect. In the second experiment, COCs were cultured singly or in different group sizes for 72 h by suitable oocyte density according to the reproductive cycle of the donor. In the anoestrous group, 1, 5 and 10 COCs were cultured in 10, 50 and 100 microl drops of the medium (10 microl per COC), respectively. In the dioestrous group, 1, 5 and 15 COCs were cultured in 7, 35 and 105 microl drops of the medium (7 microl per COC), respectively. There were no differences in the proportions of oocytes reaching metaphase II among the different group sizes in each stage of the reproductive cycle of the donor. The results indicate that the influence of oocyte density on the meiotic competence of oocytes differs according to the stage of the reproductive cycle of the donor. Moreover, the group sizes have no effect on the meiotic competence of oocytes cultured at suitable oocyte density according to the reproductive cycle of the donor. PMID- 12530916 TI - Phenotypic modulation of fibroblastic cells in the mucous layer of the human uterine cervix at term. AB - The uterine cervix is a dynamic structure with a high capacity to adapt to different, even opposing, roles during the sequence of physiological events of gestation (for example, acting as a barrier to retain the fetus during pregnancy and dilating to allow delivery at term). Histoarchitectural changes of the uterine cervix allow its successful adaptation. The aim of this study was to investigate whether fibroblastic cell plasticity, described in the lamina propria of the rat uterine cervix at term, could be observed in women too. Biopsy specimens of non-pregnant and intrapartum human cervices were studied under the transmission electron microscope, and cytoskeletal differentiation markers were identified by immunohistochemistry under the light microscope. Desmin-positive cells were present in the mucous layer of the cervix during labour. These cells displayed cytoplasmic processes (typical of myofibroblasts) that also stained positively for vimentin. The main ultrastructural features for defining the myofibroblast under the electron microscope were also observed in these cells. However, cervices of non-pregnant women contained resident fibroblasts at the same location. Examination of the differentiation repertoire of fibroblastic cells in the mucous layer of the uterine cervix resulted in the characterization of myofibroblasts at term. The implications of the plasticity of fibroblastic myofibroblastic cells in the physiological changes displayed in the uterine cervix during pregnancy, labour and postpartum involution require further investigation. PMID- 12530917 TI - Maintenance of adult mouse type A spermatogonia in vitro: influence of serum and growth factors and comparison with prepubertal spermatogonial cell culture. AB - The culture of spermatogonial cells under well-defined conditions would be an important method for elucidating the mechanisms involved in spermatogenesis and in establishing tissue regeneration in vivo. In this study, a serum-free culture system was established, with type A spermatogonia isolated from adult vitamin A deficient mice. At days 1, 3 and 7 of culture, the viability and proliferation of cells were monitored. The viability of the cells decreased by day 7 to 10% of the cells present. Proliferation occurred mainly during day 1, when 1% of the germ cells was proliferating. Co-labelling for a germ cell marker (heat shock protein 90alpha, Hsp90alpha), and a marker used to detect dividing cells (bromodeoxyuridine, BrdU), showed that this proliferation was restricted to germ cells. In an attempt to improve these parameters, medium containing fetal calf serum (FCS) was used. Viability was not influenced by serum, but proliferation was markedly enhanced. However, after day 7 of incubation with FCS, co immunolocalization for Hsp90alpha and BrdU showed a preferential proliferation of somatic cells. Comparison of cultures of adult cells with cultures of prepubertal germ cells, commonly used in studies of spermatogenesis, showed that prepubertal germ cells are twice as viable. In addition, a different proliferation profile was observed, with a peak at day 3. Here, a distinct proliferation of somatic cells was also noted. The results from the present study indicate that the origin of isolated germ cells partly determines culture outcome and that cultures of prepubertal germ cells may not be representative for adult spermatogenesis. Moreover, adding FCS to the culture medium invokes the risk of profound and undesirable effects on cell composition, also underlining the need for identification of germ cells during culture. PMID- 12530918 TI - Ovarian modulators of 11beta-hydroxysteroid dehydrogenase (11betaHSD) activity in follicular fluid from gonadotrophin-stimulated assisted conception cycles. AB - In the ovary, cortisol-cortisone interconversion is catalysed by isoforms of 11beta-hydroxysteroid dehydrogenase (11betaHSD). The objective of this study was to establish whether human follicular fluid (hFF), obtained after controlled ovarian hyperstimulation, contains paracrine modulators of 11betaHSD activity. Of 274 hFF samples tested for effects in rat kidney homogenates, 206 hFF samples significantly inhibited NADP(+)-dependent oxidation of cortisol within 1 h (by 11 67% of control 11betaHSD activity), whereas 42 hFF samples significantly stimulated 11betaHSD activity (16-210% increase relative to control). Although charcoal-stripping of hFF prevented the inhibition and potentiated the stimulation of NADP(+)-dependent cortisol oxidation in a renal homogenate, effects of individual hFF samples on NADP(+)-dependent cortisol oxidation were independent of intrafollicular progesterone concentrations. Hydrophilic fractions of hFF samples, isolated by C18 column chromatography, stimulated both the NADP(+)-dependent oxidation of cortisol (by 55+/-5%, n=98) and the NADPH dependent reduction of cortisone (by 86+/-22%, n= 5). In contrast, the hydrophobic fractions of hFF (eluted at 65-85% methanol) inhibited both NADP(+) dependent 11beta-dehydrogenase and NADPH-dependent 11-ketosteroid reductase activities (by 63+/-2% and 74+/-4%, respectively). None of the C18 column fractions of 50 hFF samples had any significant effect on NAD(+)-dependent 11beta dehydrogenase activities. The hydrophobic inhibitors of NADP(H)-dependent cortisol-cortisone metabolism did not co-elute with several candidate compounds (prostaglandins E(2) and F(2alpha), cortisol, cortisone, oestradiol, testosterone, progesterone, pregnenolone or cholesterol). Hence, hFF aspirated from women undergoing controlled ovarian hyperstimulation for assisted conception contains both hydrophilic stimuli and hydrophobic inhibitors of glucocorticoid metabolism which appear to be selective for the NADP(H)-dependent, type 1 isoform of 11betaHSD. PMID- 12530919 TI - Analysis of DNA fragmentation in bovine somatic nuclear transfer embryos using TUNEL. AB - The production of cloned animals is an inefficient process because of early or late embryonic losses. This study focused on the DNA fragmentation that occurs during embryonic development. The occurrence of DNA fragmentation was examined in bovine embryos produced by in vitro fertilization (IVF) and somatic cell nuclear transfer (NT) using the terminal deoxynucleotidyl transferase (TdT) nick-end labelling (TUNEL). IVF and NT embryos at the two-cell to blastocyst stage were stained by TUNEL for the analysis of DNA-fragmented nuclei and with propidium iodide for determination of the total number of cells. DNA fragmentation was first detected in NT embryos at the four-cell stage, but in IVF embryos at the six- to eight-cell stage. The percentage of embryos with at least one DNA fragmented nucleus increased with the advance of the developmental stage of embryos in both IVF and NT groups. The DNA-fragmented nucleus index in NT embryos that developed beyond the four-cell stage was significantly higher (P<0.01) than that of IVF embryos at the same stage. In the both IVF and NT groups, TUNEL labelled cells were detected in almost all blastocysts and were mainly observed in presumptive inner cell mass (ICM) cells of embryos. The DNA-fragmented nucleus index was negatively correlated with the total number of cells in NT blastocysts, but this relationship was not observed in IVF blastocysts. These results suggest that the high occurrence of DNA fragmentation observed in NT embryos may be related to early embryonic loss after transfer. PMID- 12530920 TI - Epididymal lithiasis in roosters and efferent ductule and testicular damage. AB - Epididymal stones have been reported in roosters in the USA and Japan. The cause of this dysfunction, which is associated with low fertility, is not known. The hypothesis of the present study is that a potential cause is the aggressive selection of birds over many centuries based upon female egg laying traits, without concern for potential effects on the male. If this hypothesis is correct, one potential consequence would be the presence of epididymal stones only in domesticated fowl and this observation would be worldwide in distribution. The present study investigated epididymal lithiasis in Brazilian crossbreed roosters and two other fowl strains, in addition to several domestic and wild bird species. The efferent ductules contained stones in 94.3% of the roosters, but stones were absent in all other domestic and wild birds. The stones were irregular in shape, size and colour and consisted mainly of calcium. In affected roosters, the efferent ductules showed epithelial cell vacuolization and sloughing and peritubular mononuclear cell infiltration, culminating with atrophy. Signs of epithelial re-canalization were seen in ductules occluded by abnormal content, such as stones. In the testis, decrease in mass, sloughing of epithelium, mononuclear cell infiltration and tubular atrophy occurred. No correlation was found between the occurrence of stones and a positive test for ELISA IBV (infectious bronchitis virus), or between the number of stones and calcium concentration in water and food, indicating that IBV infection and calcium in the diet were not related to stones formation. This study confirms and extends information about the epididymal lithiasis, which appears to be unique for roosters but to occur around the world. The severity of the lesion points to potentially severe economical impact in the poultry industry. PMID- 12530921 TI - Fertilization and activation currents in bovine oocytes. AB - One of the first events that occurs at fertilization is a transient modification of the electrical properties of the oocyte plasma membrane. The whole-cell voltage clamp technique was used to demonstrate an outward ion current and a hyperpolarization of the plasma membrane after fertilization in bovine oocytes. These electrical events, together with measurement of internal calcium concentrations, were also recorded after injection with sperm factor and exposure to parthenogenetic activators, such as Ca(2+) ionophore, ethanol and thapsigargin. Experiments were carried out simultaneously in immature and in vitro matured oocytes. Significant differences were recorded in the activation current and hyperpolarization among oocyte activators and between immature and matured oocytes. However, outward ion current and Ca(2+) release showed similar dynamics. The injection of the calcium chelator EGTA completely abolished both ion current and hyperpolarization, indicating that these electrical events are calcium dependent. Addition of specific calcium releasers, such as 1,4,5-inositol trisphosphate (IP(3)) and caffeine, triggered ion activation current and hyperpolarization indicating that IP(3) and ryanodine receptors are active in both immature and matured oocytes. Different ion channel inhibitors were used to characterize the channels underlying outward currents. Only addition of rIberiotoxin caused a complete inhibition of the current, indicating the involvement of high conductance Ca(2+)-activated K(+) channels in generating activation current. In conclusion, these findings provide evidence that bovine oocyte activation is associated with Ca(2+)-dependent electrical events. Oocytes have the potential to react to different activators even when immature; however, oocyte maturation seems to increase sensitivity to physiological activators, such as spermatozoa and sperm factor, and chemicals, such as ethanol. PMID- 12530922 TI - Quantitative (stereological) study of the effects of vasectomy on spermatogenesis in rhesus monkeys (Macaca mulatta). AB - Vasectomy reversal by vasovasostomy after long-term vasectomy in men results in lower sperm counts and pregnancy rates compared with controls, and severe damage to spermatogenesis has been observed in some animal models such as mice. The primary aim of this study was to evaluate, using sophisticated stereological methods, whether vasectomy of 6 and 12 months in a non-human primate would lead to, among other morphometric changes, reduced numbers of germ cells in testes and spermatozoa in epididymides. Five normal adult male rhesus monkeys (Macaca mulatta) underwent bilateral vasectomy, with another three aged-matched normal monkeys not undergoing vasectomy. One testis together with the ipsilateral epididymis was removed from each animal at 6 months, and the other testis and epididymis, the prostate gland and seminal vesicles were removed at 12 months. Various morphometric data were obtained using stereological methods and an unbiased and efficient stereological tool, the optical disector, was used to estimate nuclear numbers of all types of spermatogenic cells in testes and spermatozoa in epididymides using methacrylate-embedded sections 25 microm in thickness. As shown by a two-way repeated measures analysis of variance, vasectomy or hemicastration (removal of the organs at 6 months) had no significant effects on all quantitative parameters of stereology obtained from the testis, epididymis, prostate gland and seminal vesicle, except that (i) sperm granuloma was observed from three of five vasectomized animals both at 6 and 12 months, and (ii) hemicastration significantly reduced the diameter of the seminiferous tubules and increased the number of type A spermatogonia per testis. In conclusion, vasectomy in the non-human primate is a safe procedure in terms of effects on the structures of the reproductive organs. PMID- 12530924 TI - Effect of an anti-androgen on testicular descent and inguinal closure in a marsupial, the tammar wallaby (Macropus eugenii). AB - Androgens are essential for testicular descent in eutherian mammals, but little is known about its hormonal control in marsupials. This study reports the effects of daily treatment with the anti-androgen flutamide (10 mg kg(-1)) from day 9 to day 75 after birth on the descent of the testis and inguinal closure in tammar wallabies. By day 75 after birth, the testes of control males had descended and the prostate gland was well developed. The testes of all flutamide-treated males had passed through the inguinal canal and were situated in the base of the scrotum. Three of the nine flutamide-treated males had unilateral inguinal hernias. The size of the inguinal canal, regardless of whether a hernia was present, was significantly wider than that of control males. Development of the prostate gland was significantly inhibited. By day 75 after birth, the phallus was significantly longer in control males than in females, whereas the phallus of flutamide-treated males was similar to that of control females. In flutamide treated males, the lumbar 1 dorsal root ganglia was feminized and significantly fewer cell bodies expressed calcitonin gene- related peptide. As the anti androgen treatment resulted in a reduction in the number of calcitonin gene related peptide-positive cell bodies in the dorsal root ganglion supplying the genitofemoral nerve, the process of inguinal closure in tammar wallabies may be mediated by calcitonin gene-related peptide via the genitofemoral nerve, as indicated in humans. Flutamide treatment inhibited development of the prostate gland and phallus, which are both androgen-dependent structures, but it did not affect the normal descent of the testis, indicating that testicular descent can proceed when the action of androgens is blocked. PMID- 12530923 TI - Induction of capacitation and the acrosome reaction of boar spermatozoa by L arginine and nitric oxide synthesis associated with the anion transport system. AB - The aim of this study was to determine the effect of L-arginine on nitric oxide (NO) synthesis, capacitation and acrosome reaction of boar spermatozoa. Ejaculated boar spermatozoa were washed and then cultured in a bicarbonate:CO(2) buffered medium, modified NCSU-37, for 2 h. At the end of the culture, the status of spermatozoa was determined. The presence of (0.1-2.0 mmol l(-1)) L-arginine in the culture medium induced an acrosome reaction as determined by fluorescein isothiocyanate-peanut agglutinin (FITC-PNA) and increased intracellular NO content, as quantified by a fluorescent indicator, diaminofluorescein-2 diacetate (DAF-2 DA). This stimulatory effect of L-arginine was neutralized by supplementation with an NO synthase inhibitor, N(omega)-nitro-L-arginine methyl ester (1 mmol l(-1)). However, the inactive enantiomorph, N(omega)-nitro-D arginine methyl ester, did not affect the stimulatory effect of L-arginine. These results indicate that L-arginine induces an acrosome reaction through the NO signal pathway in boar spermatozoa. Furthermore, the stimulatory effect of L arginine was inhibited in the presence of an anion transport inhibitor, 4 acetamido-4'-isothiocyano-stilbene-2,2'-disulphonic acid (SITS; 0.1 mmol l(-1)), whereas any responses of spermatozoa to caffeine were not inhibited by SITS. A stimulatory effect of L-arginine on capacitation and acrosome reaction of spermatozoa was also observed in modified NCSU37 medium by using a chlortetracycline fluorescence assay, but not in supplemented bicarbonate-free Tris-buffered medium. These results indicate that the presence of L-arginine induces nitric oxide synthesis and stimulates capacitation and acrosome reaction of boar spermatozoa only when active sperm anion transport is present as a result of bicarbonate supplementation. PMID- 12530925 TI - Follicular waves and circulating concentrations of gonadotrophins, inhibin and oestradiol during the anovulatory season in mares. AB - Follicular waves and associated circulating hormone concentrations were studied during the anovulatory season in pony mares (n=8). Follicles were monitored by ultrasonography and a blood sample was taken daily from 29 January until ovulation (mean, 28 April). A mid-anovulatory period (largest follicle, 16.0+/ 0.5 mm in diameter) and transitional period (largest follicle, 22.4+/-0.5 mm) were distinctive in each mare. The two periods were delineated by an increase in the diameter of the largest follicle to >/=21.0 mm. Follicular waves, identified by significant increases in the mean diameter of the second to sixth largest follicles, were detected during both the mid-anovulatory and transitional periods. The mean number of follicles >/=15.0 mm in diameter and the diameter of the second to sixth largest follicles increased in association with statistically identified FSH surges. The pattern of the FSH concentration changes during surges did not change during the mid-anovulatory and transitional periods. During the declining portion of the FSH surge, follicle growth continued and circulating total inhibin increased, indicating suppression of FSH by inhibin from the growing follicles. Circulating oestradiol or LH did not change relative to wave emergence. Results indicated that follicular waves occurred during the second half of the anovulatory season, even during the period of lowest follicular activity. On a temporal basis, follicular wave emergence was stimulated by surges in circulating FSH. However, the increase in follicle growth to >/=21.0 mm in diameter for the wave at the beginning of the transitional period and for the subsequent waves was not attributable to a change in the characteristics of the associated FSH surges. PMID- 12530926 TI - Mediation of contraction in rat cauda epididymidis by alpha-adrenoceptors. AB - Subtypes of alpha-adrenoceptors responsible for contractions of the rat cauda epididymidis were studied in vivo by micropuncture using a servo-nulling pressure transducer system. Administration of both non-selective and selective alpha adrenoceptor agonists in doses of 1-40 microg noradrenaline kg(-1) body weight (BW), 1-100 microg clonidine kg(-1) BW, or 100-800 mg methoxamine kg(-1) BW enhanced contractions of the proximal cauda epididymidis in a dose-response manner. The potency of the agonists were noradrenaline > or = clonidine>methoxamine. Pre-treatments with selective alpha(1)-adrenoceptor antagonist (prazosin) and alpha(2)-adrenoceptor antagonist (yohimbine) at the doses of 400 and 800 microg kg(-1) BW, respectively, had very little effect on spontaneous contractions, but effectively blocked the responses to the maximal doses of methoxamine and clonidine. The responses could not be explained by the systemic effects of agonists and antagonists. The results suggest that contraction of the proximal cauda epididymidis of rats is mediated by both alpha(1)- and alpha(2)-adrenoceptors. The latter appears to be more abundant. PMID- 12530928 TI - Role of lipoxygenase metabolites in cardiovascular disease. PMID- 12530927 TI - Phage annealing proteins promote oligonucleotide-directed mutagenesis in Escherichia coli and mouse ES cells. AB - BACKGROUND: The phage protein pairs, RecE/RecT from Rac or Redalpha/Redbeta from lambda, initiate efficient double strand break repair (DSBR) in Escherichia coli that has proven very useful for DNA engineering. These phage pairs initiate DSBR either by annealing or by another mechanism that is not defined. RESULTS: Here we report that these proteins also mediate single strand oligonucleotide repair (ssOR) at high efficiencies. The ssOR activity, unlike DSBR, does not require a phage exonuclease (RecE or Redalpha) but only requires a phage annealing protein (RecT or Redbeta). Notably, the P22 phage annealing protein Erf, which does not mediate the same DSBR reactions, also delivers ssOR activity. By altering aspects of the oligonucleotides, we document length and design parameters that affect ssOR efficiency to show a simple relationship to homologies either side of the repair site. Notably, ssOR shows strand bias. Oligonucleotides that can prime lagging strand replication deliver more ssOR than their leading complements. This suggests a model in which the annealing proteins hybridize the oligonucleotides to single stranded regions near the replication fork. We also show that ssOR is a highly efficient way to engineer BACs and can be detected in a eukaryotic cell upon expression of a phage annealing protein. CONCLUSION: Phage annealing proteins can initiate the recombination of single stranded oligonucleotides into endogenous targets in Escherichia coli at very high efficiencies. This expands the repertoire of useful DNA engineering strategies, shows promise for applications in eukaryotic cells, and has implications for the unanswered questions regarding DSBR mediated by RecE/RecT and Redalpha/Redbeta. PMID- 12530929 TI - Best bang for the buck? PMID- 12530930 TI - Epithelial sodium channel, salt intake, and hypertension. AB - The epithelial sodium channel (ENaC) is a membrane protein made of three different but homologous subunits (a, b, and g) present in the apical membrane of epithelial cells of, for example, the distal nephron. This channel is responsible for salt reabsorption in the kidney and can cause human diseases by increasing channel function in Liddle's syndrome, a form of hereditary hypertension, or by decreasing channel function in pseudohypoaldosteronism type I, a salt-wasting disease in infancy. This review briefly discusses recent advances in understanding the implication of ENaC in Liddle's syndrome and in pseudohypoaldosteronism type I, both caused by mutations in the SCNN1 (ENaC) genes. Furthermore, it is still an open question to which extent SCNN1 genes coding for ENaC might be implicated in essential hypertension. The development of Scnn1 genetically engineered mouse models will provide the opportunity to test the effect of environmental factors, like salt intake, on the development of this kind of salt- sensitive hypertension. PMID- 12530931 TI - Endothelial nitric oxide synthase polymorphisms and hypertension. AB - The human endothelial nitric oxide synthase (eNOS) gene is highly polymorphic. Evidence for the involvement of eNOS single nucleotide polymorphisms in the development of essential hypertension is limited, though the eNOS Glu298Asp polymorphism appears to influence the blood pressure response to exercise. This variant also influences endothelial function, with its effects becoming manifest during the adaptive vascular changes of pregnancy. Carriers of eNOS Asp298 may be at risk of developing pre-eclampsia. Molecular studies have indicated that intact eNOS Asp298 has equivalent enzymatic activity to eNOS Glu298, but undergoes selective proteolysis in native cells and tissues such that the steady state level of active eNOS may be reduced in carriers of this allele. Carriers of eNOS Asp298, particularly if exposed to adverse environmental infuences on endothelial function, may be at increased risk of developing atherosclerosis and cerebrovascular disease. PMID- 12530932 TI - Homocysteine and blood pressure. AB - Several studies, some population-based, have linked plasma homocysteine levels to blood pressure, especially systolic pressure. The strength of this association is weak, but may be underestimated due to inaccurate blood pressure measurements. In addition, the association may be confounded by renal function. Observations that homocysteine-lowering therapies with folic acid-based treatments have been followed by decreases in blood pressure, however, raise the possibility that the link between homocysteine and blood pressure is real, which is important as homocysteine levels can easily be lowered by folic acid-based regimens. Mechanisms that could explain the relationship between homocysteine and blood pressure include increased arterial stiffness, endothelial dysfunction with decreased availability of nitric oxide, low folate status, and insulin resistance. So far, however, no evidence has been provided that these mechanisms are operative in humans. Ongoing large intervention studies with homocysteine lowering vitamins may indicate whether blood pressure is indeed lowered by these vitamins, whether the blood pressure decrease, if any, is explained by the decrease in homocysteine levels, and whether a vitamin treatment-associated decrease in cardiovascular morbidity (if any) is explained by the decrease in blood pressure. PMID- 12530933 TI - Air pollution: the "Heart" of the problem. AB - Air pollution exposure is associated with an increased risk of acute and chronic cardiovascular mortality. Recent observations have implicated fine particulate matter (PM(2.5)) as one of the most important pollutants. Inhalation of PM(2.5) causes acute pulmonary inflammation and oxidative stress. The subsequent generation of a systemic inflammatory response could link air pollution exposure with the development of cardiovascular disease. Human experiments have demonstrated pro-arrhythmic alterations in cardiac autonomic tone, increased blood pressure, higher serum C-reactive protein levels, and alterations in blood rheology favoring coagulation following controlled pollution exposures or in relation to elevated ambient PM(2.5) levels. Recent studies have also uncovered several harmful impacts on the systemic vasculature, including the triggering of acute vasoconstriction and the enhanced development of atherosclerosis. Many questions, however, remain unanswered and future studies will be required to clarify the relevant biologic mechanisms and to identify the specific constituents responsible for mediating the adverse health impacts. PMID- 12530934 TI - Linkage disequilibrium analysis of the renin-angiotensin system genes. AB - The genes of the renin-angiotensin system (RAS) are important candidates to confer susceptibility to cardiovascular diseases. A large number of association studies between cardiovascular traits and the polymorphisms in RAS have been conducted, although inconsistent results are often reported. The patterns of linkage disequilibrium in RAS genes have also been reported in different populations. However, our understanding of the genetic architecture underlying the RAS is still limited despite rapid progress in empiric studies regarding the patterns of the human genome as a whole. In this review, the linkage disequilibrium among the polymorphisms within the four RAS genes and current association analyses involving the RAS are discussed, as well as some of the gaps of knowledge and possible solutions. PMID- 12530936 TI - Comparison of omapatrilat and enalapril in patients with chronic heart failure: the Omapatrilat Versus Enalapril Randomized Trial of Utility in Reducing Events (OVERTURE). PMID- 12530935 TI - G-protein beta3 subunit 825T allele and hypertension. AB - The G-protein beta3 subunit (GNB3) C825T polymorphism was detected through a classical candidate gene approach using cell lines with enhanced G-protein activation from patients with essential hypertension. The 825T allele is associated with the expression of a shortened, functionally active splice variant of the G-protein beta3 subunit and enhanced intracellular signal transduction. Independent studies have confirmed an association of the 825T allele with hypertension in whites. Potential pathogenetic mechanisms comprise an increased susceptibility for obesity in 825T allele carriers and, potentially, increased responsiveness to vasoactive hormones. Both phenomena appear to be strongly influenced by lifestyle in the sense of a gene-environment interaction. Whether hypertensive 825T allele carriers are at increased risk for stroke and left ventricular hypertrophy remains controversial. Current studies try to define optimal therapy strategies for hypertensive 825T allele carriers. PMID- 12530937 TI - Aldosterone: a risk factor for vascular disease. AB - Blockade of the renin-angiotensin-aldosterone system with angiotensin-converting enzyme (ACE) inhibitors or angiotensin receptor antagonists has resulted in beneficial effects in essential hypertensive patients. However, occurrence of cardiovascular events has not been appropriately controlled beyond a certain percentage. One reason could be the effects of aldosterone, the final component of the system. The aldosterone escape phenomenon could explain undesirable outcomes observed in hypertensive patients even under treatment with ACE inhibitors or angiotensin antagonists. Aldosterone has direct effects on the vasculature and has been associated with vascular smooth muscle cell hypertrophy, endothelial dysfunction, cardiac fibrosis, proteinuria, and renal vascular injury. Animal models and clinical trials have proven the benefit of aldosterone receptor antagonism. With increased recognition of the prevalence of hyperaldosteronism in patients thought to have "essential" hypertension, the use of drugs that block aldosterone action may become more widespread and protect the vasculature from the deleterious effects of aldosterone. PMID- 12530938 TI - The role of RhoA and Rho-associated kinase in vascular smooth muscle contraction. AB - A variety of contractile agonists trigger activation of the small GTPase RhoA. An important target of activated RhoA in smooth muscle is Rho-associated kinase (ROK), one of the downstream targets that is the myosin binding subunit (MYPT1) of myosin light chain phosphatase (MLCP). Phosphorylation of MYPT1 at T695 by activated ROK results in a decrease in phosphatase activity of MLCP and an increase in myosin light chain (LC(20)) phosphorylation catalyzed by Ca(2)(+)/calmodulin-dependent myosin light chain kinase and/or a distinct Ca(2)(+)-independent kinase. LC(20) phosphorylation in turn triggers cross-bridge cycling and force development. ROK also phosphorylates the cytosolic protein CPI 17 (at T38), which thereby becomes a potent inhibitor of MLCP. The RhoA/ROK pathway has been implicated in the tonic phase of force maintenance in response to various agonists, with no evident role in the phasic response, suggesting this pathway as a potential target for antihypertensive therapy. Indeed, ROK inhibitors restore normal blood pressure in several rat hypertensive models. PMID- 12530939 TI - Molecular mechanisms of angiotensin II-induced vascular injury. AB - Blockers of the renin-angiotensin system are used in the treatment of several cardiovascular and renal diseases, including hypertension, atherosclerosis, and cardiac failure. Angiotensin II plays an essential role in the pathogenesis of these diseases through the regulation of cell growth, inflammation, and fibrosis. There are two main angiotensin II receptors, AT(1) and AT(2). The AT(1) receptor is responsible for most of the pathophysiologic actions of angiotensin II, including cell proliferation, production of growth factors and cytokines, and fibrosis. AT(2) causes antiproliferation and counteracts the cell growth induced by AT(1) activation. We review the mechanisms whereby AT(1) and AT(2) receptors elicit their respective actions. We discuss the current understanding of the signaling mechanisms involved in angiotensin II-induced vascular damage, describing the mediators (growth factors and cytokines) and intracellular signals (activation of protein kinases, transcription factors, and redox pathways) implicated in these processes, with special emphasis on novel information and open questions. PMID- 12530942 TI - Treatment for chronic hepatitis C. PMID- 12530940 TI - The L-arginine-nitric oxide pathway in hypertension. AB - Nitric oxide is involved in the regulation of resting vascular tone, adaptation of blood flow to metabolic demand of tissue, and adaptation of vessel diameter to volume of inflow, ie, flow-mediated dilation. Arterial hypertension is associated with an increased vascular tone of resistance vessels, a reduced compliance of conduit arteries, along with a thickening of the intima-media leading to vascular remodeling. Dysfunctional endothelium triggers such maladaptive processes. A reduced bioavailability of nitric oxide has been shown in hypertensive individuals dependent on the duration and severity of arterial hypertension. Angiotensin-converting enzyme inhibitors reverse endothelial dysfunction, whereas a concomitant reduction in significant cardiac events due to improved bioavailability has yet to be established. Long-term follow-up studies in individuals with manifest endothelial dysfunction and in offspring from hypertensive patients underscore the prognostic and genetic significance of a reduced nitric oxide bioavailability for the pathophysiology of arterial hypertension. PMID- 12530941 TI - Does cyclooxygenase-2 affect blood pressure? AB - With the development and clinical implementation of the new cyclooxygenase (COX) 2 inhibitors, their safety, including the effects on renal function and blood pressure, is attracting increasing attention. In the kidney, COX-2 is constitutively expressed and is highly regulated in response to alterations in intravascular volume. COX-2 metabolites have been implicated in mediation of renin release, regulation of sodium excretion, and maintenance of renal blood flow. Similar to conventional nonsteroidal anti-inflammatory drugs, inhibition of COX-2 may cause modest elevations in blood pressure in a minority of subjects. COX-2 inhibitors may also exacerbate pre-existing hypertension or interfere with other antihypertensive drugs. Special caution should be taken in patients with volume depletion or decreased organ perfusion. PMID- 12530943 TI - The management of primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) is a chronic cholestatic hepatobiliary disease that usually progresses to biliary cirrhosis and liver failure; it also predisposes to cholangiocarcinoma. The cause of PSC is unknown, although evidence suggests that the tissue damage is mediated by the immune system. There is an unexplained close association between PSC and inflammatory bowel disease, particularly in ulcerative colitis, which coexists in the majority of patients with PSC. No medical therapy has been proven to halt or reverse disease progression; however, recent preliminary evidence suggests that ursodeoxycholic acid (UDCA) in a high dose of 20 to 25 mg/kg may slow the disease process. Evidence from a pilot study suggests that the combination of UDCA and immunosuppressive therapy, such as prednisolone or azathioprine, may also increase efficacy. For patients with end-stage PSC, liver transplantation remains the only effective therapy, although there is clear evidence that PSC may recur in the liver allograft. PMID- 12530945 TI - Ethical issues in living donor liver transplantation. AB - The cadaveric organ shortage and the high mortality rate while patients wait for an organ have driven the medical community to develop alternative strategies for treating patients with end-stage liver disease. Adult living donor liver transplantation (ALDT) has evolved in response to the cadaveric organ shortage. Although there are benefits for recipients of ALDT, donors may incur substantial risk, including death. In contrast to pediatric living donation, in which the left lateral segment of the liver is resected from a donor, ALDT generally requires right hepatectomy, which is associated with greater morbidity and mortality. Because ALDT places a healthy individual at risk for substantial morbidity and mortality, debate over the ethics of this procedure is ongoing. Two donor deaths have occurred in the United States, adding to the concern over donor safety. Despite the risks associated with ALDT, many individuals elect to proceed with living donation with the hope of improving the life of a relative or friend. When considering whether we as a society should support and encourage ALDT, we should examine the perspective of the donor, recipient, and medical community as well. The medical community has an obligation to study carefully the risks and outcomes associated with ALDT so that we can deliver the highest quality of care that is not at the expense of healthy individuals. PMID- 12530944 TI - Interaction of iron, insulin resistance, and nonalcoholic steatohepatitis. AB - Nonalcoholic fatty liver disease (NAFLD) has emerged as a ubiquitous liver disorder with occasional serious overtones. Although diabetes and obesity were initially held culpable, insulin resistance (IR) is now considered the fundamental operative mechanism. IR is probably the "first step" in nonalcoholic steatohepatitis (NASH). Oxidative stress may be the elusive "second" of possibly multiple steps in the progression of steatosis to fibrosing steatohepatitis. Because hepatic iron promotes oxidative stress, it was mooted as a contributory cofactor in NASH. This proposal was strengthened by an association with hepatic fibrosis. Subsequent studies have shown neither a significant increase in hepatic iron nor an association between hepatic iron and any of the histologic determinants in NASH. Likewise, the increased prevalence of hemochromatosis gene (HFE) mutations in some studies appears to be largely irrelevant to the development of hepatic fibrosis. Excess hepatic iron may occur in insulin resistance-associated iron overload (IRHIO), characterized by hyperferritinemia with normal to mild increases in transferrin saturation. Although patients with IRHIO have a high prevalence of IR-related metabolic disorders, the relationship of IRHIO to NASH is unclear. A recent study showed improvement in insulin sensitivity with the use of venesection in patients with NAFLD, but this approach cannot be implemented without extensive review. PMID- 12530946 TI - Current use of transjugular intrahepatic portosystemic shunts. AB - The principal indication for transjugular intrahepatic portosystemic shunts (TIPS) continues to be rescue therapy for variceal hemorrhage that cannot be controlled by endoscopic or medical therapy. TIPS provide no survival advantage in prevention of rebleeding or refractory ascites. The indications for TIPS continue to expand, however, especially for Budd-Chiari syndrome and hydrothorax. Other more novel indications include bleeding portal hypertensive gastropathy or ectopic varices, Budd-Chiari syndrome, veno-occlusive disease, hepatorenal syndrome, hepatopulmonary syndrome, hepatocellular carcinoma, and polycystic liver disease. Great strides have been made recently in models to predict mortality and complications following TIPS placement. Graft stents hold promise based on early studies. Finally, complications are common and may be life threatening. PMID- 12530947 TI - Etiology and management of fulminant hepatic failure. AB - Fulminant hepatic failure (FHF) remains a rare but devastating disease. Viruses and drug-induced hepatotoxicity are the most common causes of the syndrome, but the relevance of each differs depending on the geographic area. In a large proportion of patients no cause for FHF can be identified. Good intensive care is critical for patient survival. Orthotopic liver transplantation (OLT) remains a definitive therapeutic option. Prognostic indices have helped to optimize patient selection and timing for performance of OLT. However, the accuracy of these prognostic indices decreases when they are applied to different populations, probably because of regional differences in etiology and peculiar native host factors. More accurate prognostic criteria and new therapeutic alternatives to OLT are required. PMID- 12530948 TI - Treatment of hepatic fibrosis: almost there. AB - Hepatic fibrosis is the scarring response of the liver to chronic liver injury; when fibrosis progresses to cirrhosis, morbid complications can develop. Available therapies for many chronic liver diseases are ineffective, with liver transplantation as the only option, though the supply of donor organs is inadequate to meet the growing demand. Novel approaches that attack the scarring response are therefore urgently needed. Optimism in this effort is fueled by major insights into the pathogenesis of fibrosis and by accumulating evidence that even cirrhosis is reversible in many patients. Most evolving antifibrotic therapies will be aimed at inhibiting the activated hepatic stellate cell, which is responsible for the fibrotic response to injury. This review describes the ways in which insights into the cellular basis of hepatic fibrosis are leading to realistic strategies for antifibrotic treatment that may revolutionize the management of patients with chronic liver disease. PMID- 12530949 TI - Screening for hepatocellular carcinoma: why, when, how? AB - Hepatocellular carcinoma (HCC) is a major public health concern in many areas of the world, and its incidence is increasing in the United States and other countries. Screening for HCC in patients with cirrhosis has been advocated to identify those with small lesions who would benefit from transplantation or surgical resection. Despite these recommendations, several issues regarding screening remain controversial. No randomized, controlled trials have confirmed that surveillance for HCC reduces disease-specific mortality. In addition, the most appropriate screening test and optimal screening interval have not yet been defined. Clearly, these unresolved questions have a major impact on the cost effectiveness of a screening program either at the population or the clinic level. A few studies, however, have suggested that screening may be cost effective because a minor survival benefit could result in a cost that is acceptable to decision makers. PMID- 12530950 TI - Vascular liver diseases. AB - This article reviews the primary circulatory liver diseases, which include Budd Chiari syndrome, obstruction of the hepatic portion of the inferior vena cava, portal vein thrombosis, sinusoidal obstruction syndrome (veno-occlusive disease), nodular regenerative hyperplasia, and peliosis hepatis. In addition, two systemic cardiovascular diseases that impair hepatic circulation, ischemic hepatitis and congestive hepatopathy, are briefly discussed. A characteristic of the primary circulatory liver diseases is that portal hypertension usually precedes liver dysfunction; however, this is not the case with the primary parenchymal liver diseases, in which liver dysfunction always progresses before portal hypertension is manifested. Significant overlap exists among the diseases and risk factors that predispose patients to the primary circulatory liver diseases, though the pathogenesis of individual diseases varies. PMID- 12530951 TI - Metabolic bone disease in patients with liver disease. AB - The coexistence of liver disease and osteopenic bone disease has been recognized for many years and is now the subject of increasing attention. Osteoporosis has been characterized well in patients with cholestatic liver disease, but new research suggests that osteopenia and osteoporosis may also be prevalent in patients with other chronic liver diseases. Although the precise mechanism of bone loss remains unclear, advances in treatment and prevention are bringing heightened awareness to this common problem. PMID- 12530952 TI - Treatment of chronic hepatitis C in nonresponders to previous therapy. AB - About 55% to 60% of treatment-naive patients fail to achieve a sustained virologic response after therapy with standard interferon and ribavirin. The use of polyethylene glycol-enhanced interferon (PEG-IFN) plus ribavirin for retreatment of this challenging group is currently being evaluated by several investigators. Although no sustained virologic response rates have been reported yet from these trials, reported on-treatment response rates for previous nonresponders range from 25% to 30%, and an early estimate of the sustained virologic response rate is about 11%. Outcomes for treatment of relapsers and interferon monotherapy nonresponders have been significantly better than those for combination nonresponders. On-treatment responses of 40% to 43% in previous combination therapy nonresponders are now being seen with the addition of amantadine (triple therapy), and sustained response rates with this regimen are awaited. Large trials are underway to evaluate the role of maintenance therapy for virologic nonresponders with advanced liver disease. PMID- 12530954 TI - Epithelial polarity in flies: more than just crumbs. AB - The plasma membrane of polarized epithelial cells is composed of different domains, which are associated with specific protein complexes. Recent studies in Drosophila shed new light on the functional interactions of these protein complexes during epithelial differentiation in embryogenesis. PMID- 12530953 TI - Pathogenic interactions between alcohol and hepatitis C. AB - Alcohol is the most commonly abused substance in the United States, and alcohol abuse leads to alcoholic liver disease, a long recognized major public health concern. The high prevalence of chronic hepatitis C virus (HCV) infection, along with the clinical observation that HCV infection is common in alcoholic patients presenting with liver disease, has directed attention to the interaction between alcohol and HCV infection. Clinical studies have identified alcohol use as an independent risk factor for progression of fibrosis in chronic HCV infection. Experimental evidence suggests additive inhibitory effects between HCV and alcohol on antiviral immune responses. In addition, specific pathways have been identified by which HCV core protein and alcohol interact to activate hepatocytes. Nonspecific inflammatory cell recruitment and proinflammatory cytokine activation have also been implicated in both alcohol- and HCV-induced liver diseases. Further investigation of these and other pathways by which alcohol and HCV interact should unravel the mechanisms that accelerate the progression of liver disease. PMID- 12530955 TI - The immunological synapse and actin assembly: a regulatory role for PKC theta. AB - Engagement of the T cell receptor leads to the accumulation of filamentous actin, which is necessary for the formation of the immunological synapse and subsequent T cell activation. In the December issue of Molecular Cell, Sasahara et al. provide new insights into the link between the T cell receptor and actin assembly in the immunological synapse, and reveal a critical regulatory role for PKC theta in this process. PMID- 12530956 TI - Flies kNOw how to signal. AB - A recent study has discovered a surprising role for nitric oxide in the Drosophila immune response. NO-mediated signaling was implicated in the communication between the site of a localized infection and the major immune organ of the fly, the fat body. PMID- 12530957 TI - Bringing classical embryology to C elegans gastrulation. AB - In a recent paper, Lee and Goldstein develop an explant assay that recapitulates key aspects of gastrulation in C elegans and permits classical embryological manipulations. The resulting detailed analysis of cell behavior will ultimately extend to broader issues, such as, whether morphogenesis can be described as the sum of single-cell events or if unique phenomena emerge at the multicellular level. PMID- 12530958 TI - In search of lipid translocases and their biological functions. AB - In plasma membranes, lipids distribute asymmetrically across the bilayer, a process that requires proteins. Recent work identified novel lipid translocators in yeast, and their activity was functionally correlated to endocytosis, thus boosting investigations on identity, mechanism, and function of lipid translocases. PMID- 12530959 TI - Tube or not tube: remodeling epithelial tissues by branching morphogenesis. AB - Branching morphogenesis involves the restructuring of epithelial tissues into complex and organized ramified tubular networks. Early rounds of branching are controlled genetically in a hardwired fashion in many organs, whereas later, branching is stochastic, responding to environmental cues. We discuss this sequential process from formation of an organ anlage and invagination of the epithelium to branch initiation and outgrowth in several model systems including Drosophila trachea and mammalian lung, mammary gland, and kidney. PMID- 12530960 TI - The bacterial cytoskeleton: in vivo dynamics of the actin-like protein Mbl of Bacillus subtilis. AB - Mbl is a bacterial actin homolog that controls cell morphogenesis in Bacillus subtilis. A functional GFP-Mbl fusion protein was used to examine the behavior of the helical cables formed by Mbl protein in live B. subtilis cells. The cables undergo dynamic changes during cell cycle progression. They are stable but not rigid while elongating in parallel with cell growth, and they require septum formation to divide/cleave. Fluorescence recovery after photobleaching (FRAP) analysis showed that the cables are continuously remodeled during cell elongation. Turnover occurs along the length of the helical Mbl filaments, with no obvious polarity and a recovery half-time of about 8 min. These findings have important implications for the nature of bacterial cell wall architecture and synthesis. PMID- 12530961 TI - A RhoGEF and Rho family GTPase-activating protein complex links the contractile ring to cortical microtubules at the onset of cytokinesis. AB - The mechanism that positions the cytokinetic contractile ring is unknown, but derives from the spindle midzone. We show that an interaction between the Rho GTP exchange factor, Pebble, and the Rho family GTPase-activating protein, RacGAP50C, connects the contractile ring to cortical microtubules at the site of furrowing in D. melanogaster cells. Pebble regulates actomyosin organization, while RacGAP50C and its binding partner, the Pavarotti kinesin-like protein, regulate microtubule bundling. All three factors are required for cytokinesis. As furrowing begins, these proteins colocalize to a cortical equatorial ring. We propose that RacGAP50C-Pavarotti complexes travel on cortical microtubules to the cell equator, where they associate with the Pebble RhoGEF to position contractile ring formation and coordinate F-actin and microtubule remodeling during cytokinesis. PMID- 12530962 TI - Recognition of a bicoid mRNA localization signal by a protein complex containing Swallow, Nod, and RNA binding proteins. AB - Localization of mRNAs, a process essential for embryonic body patterning in Drosophila, requires recognition of cis-acting signals by cellular components responsible for movement and anchoring. We have purified a large multiprotein complex that binds a minimal form of the bicoid mRNA localization signal in a manner both specific and sensitive to inactivating mutations. Identified complex components include the RNA binding proteins Modulo, PABP, and Smooth, the known localization factor Swallow, and the kinesin family member Nod. We demonstrate that localization of bcd mRNA is defective in modulo mutants. The presence of three required localization components (Swallow, Modulo, and specific RNA binding activity) within the recognition complex strongly implicates it in mRNA localization. PMID- 12530964 TI - Mind bomb is a ubiquitin ligase that is essential for efficient activation of Notch signaling by Delta. AB - Lateral inhibition, mediated by Notch signaling, leads to the selection of cells that are permitted to become neurons within domains defined by proneural gene expression. Reduced lateral inhibition in zebrafish mib mutant embryos permits too many neural progenitors to differentiate as neurons. Positional cloning of mib revealed that it is a gene in the Notch pathway that encodes a RING ubiquitin ligase. Mib interacts with the intracellular domain of Delta to promote its ubiquitylation and internalization. Cell transplantation studies suggest that mib function is essential in the signaling cell for efficient activation of Notch in neighboring cells. These observations support a model for Notch activation where the Delta-Notch interaction is followed by endocytosis of Delta and transendocytosis of the Notch extracellular domain by the signaling cell. This facilitates intramembranous cleavage of the remaining Notch receptor, release of the Notch intracellular fragment, and activation of target genes in neighboring cells. PMID- 12530965 TI - Conserved requirement of Lim1 function for cell movements during gastrulation. AB - To investigate Lim1 function during gastrulation, we used transcript depletion through DEED antisense oligonucleotides in Xenopus and cell transplantation in mice. Xenopus embryos depleted of Lim1 lack anterior head structures and fail to form a proper axis as a result of a failure of gastrulation movements, even though mesodermal cell identities are specified. Similar disruption of cell movements in the mesoderm is also observed in Lim1(-/-) mice. Paraxial protocadherin (PAPC) expression is lost in the nascent mesoderm of Lim1(-/-) mouse embryos and in the organizer of Lim1-depleted Xenopus embryos; the latter can be rescued to a considerable extent by supplying PAPC exogenously. We conclude that a primary function of Lim1 in the early embryo is to enable proper cell movements during gastrulation. PMID- 12530963 TI - Two RNA binding proteins, HEN4 and HUA1, act in the processing of AGAMOUS pre mRNA in Arabidopsis thaliana. AB - AGAMOUS, a key player in floral morphogenesis, specifies reproductive organ identities and regulates the timely termination of stem cell fates in the floral meristem. Here, we report that strains carrying mutations in three genes, HUA1, HUA2, and HUA ENHANCER4 (HEN4), exhibit floral defects similar to those in agamous mutants: reproductive-to-perianth organ transformation and loss of floral determinacy. HEN4 codes for a K homology (KH) domain-containing, putative RNA binding protein that interacts with HUA1, a CCCH zinc finger RNA binding protein in the nucleus. We show that HUA1 binds AGAMOUS pre-mRNA in vitro and that HEN4, HUA1, and HUA2 act in floral morphogenesis by specifically promoting the processing of AGAMOUS pre-mRNA. Our studies underscore the importance of RNA processing in modulating plant development. PMID- 12530966 TI - Drosophila matrix metalloproteinases are required for tissue remodeling, but not embryonic development. AB - The matrix metalloproteinase (MMP) family is heavily implicated in many diseases, including cancer. The developmental functions of these genes are not clear, however, because the >20 mammalian MMPs can be functionally redundant. Drosophila melanogaster has only two MMPs, which are expressed in embryos in distinct patterns. We created mutations in both genes: Mmp1 mutants have defects in larval tracheal growth and pupal head eversion, and Mmp2 mutants have defects in larval tissue histolysis and epithelial fusion during metamorphosis; neither is required for embryonic development. Double mutants also complete embryogenesis, and these represent the first time, to our knowledge, that all MMPs have been disrupted in any organism. Thus, MMPs are not required for Drosophila embryonic development, but, rather, for tissue remodeling. PMID- 12530967 TI - Cysteine-rich LIM-only proteins CRP1 and CRP2 are potent smooth muscle differentiation cofactors. AB - Cysteine-rich LIM-only proteins, CRP1 and CRP2, expressed during cardiovascular development act as bridging molecules that associate with serum response factor and GATA proteins. SRF-CRP-GATA complexes strongly activated smooth muscle gene targets. CRP2 was found in the nucleus during early stages of coronary smooth muscle differentiation from proepicardial cells. A dominant-negative CRP2 mutant blocked proepicardial cells from differentiating into smooth muscle cells. Together with SRF and GATA proteins, CRP1 and CRP2 converted pluripotent 10T1/2 fibroblasts into smooth muscle cells, while muscle LIM protein CRP3 inhibited the conversion. Thus, LIM-only proteins of the CRP family play important roles in organizing multiprotein complexes, both in the cytoplasm, where they participate in cytoskeletal remodeling, and in the nucleus, where they strongly facilitate smooth muscle differentiation. PMID- 12530968 TI - The forkhead transcription factor Foxo1 regulates adipocyte differentiation. AB - An outstanding question in adipocyte biology is how hormonal cues are relayed to the nucleus to activate the transcriptional program that promotes adipogenesis. The forkhead transcription factor Foxo1 is regulated by insulin via Akt-dependent phosphorylation and nuclear exclusion. We show that Foxo1 is induced in the early stages of adipocyte differentiation but that its activation is delayed until the end of the clonal expansion phase. Constitutively active Foxo1 prevents the differentiation of preadipocytes, while dominant-negative Foxo1 restores adipocyte differentiation of fibroblasts from insulin receptor-deficient mice. Further, Foxo1 haploinsufficiency protects from diet-induced diabetes in mice. We propose that Foxo1 plays an important role in the integration of hormone activated signaling pathways with the complex transcriptional cascade that promotes adipocyte differentiation. PMID- 12530970 TI - Nature's TRAIL--on a path to cancer immunotherapy. AB - The TNF-related apoptosis-inducing ligand (TRAIL) offers great promise as a cancer therapeutic. Initially, soluble recombinant versions of the TRAIL molecule have exhibited specific tumoricidal activity against a variety of tumors alone, or in combination with other cancer treatments, and much anticipation awaits the outcomes from early clinical trials. More recently, the natural role of TRAIL has been explored in tumor and allogeneic bone marrow transplantation models in the mouse. Strikingly, the TRAIL effector pathway appears a vital component of immunosurveillance of spontaneous or resident tumor cells by both T cells and NK cells, stimulating more hope that manipulating TRAIL activity is a natural path to improved cancer immunotherapy. PMID- 12530969 TI - C elegans: a model for exploring the genetics of fat storage. AB - To gain insights into the genetic cascades that regulate fat biology, we evaluated C. elegans as an appropriate model organism. We generated worms that lack two transcription factors, SREBP and C/EBP, crucial for formation of mammalian fat. Worms deficient in either of these genes displayed a lipid depleted phenotype-pale, skinny, larval-arrested worms that lack fat stores. On the basis of this phenotype, we used a reverse genetic screen to identify several additional genes that play a role in worm lipid storage. Two of the genes encode components of the mitochondrial respiratory chain (MRC). When the MRC was inhibited chemically in worms or in a mammalian adipocyte model, fat accumulation was markedly reduced. A third encodes lpd-3, whose homolog is also required for fat storage in a mammalian model. These data suggest that C. elegans is a genetically tractable model to study the mechanisms that underlie the biology of fat-storing tissues. PMID- 12530971 TI - Not just any T cell receptor will do. AB - Although our structural understanding of T cell recognition has rapidly evolved due to recent crystallographic results, the reality is that detailed answers to many of the most fundamental questions still remain elusive. In this issue, high resolution insight into the phenomenon of TCR chain bias takes down another brick from the wall. PMID- 12530972 TI - Induction of the NF-kappaB cascade by recruitment of the scaffold molecule NEMO to the T cell receptor. AB - The mechanism by which TCR signaling activates NF-kappaB is poorly understood. We demonstrate here that the IKK kinase complex is recruited to the immunological synapse and can be coprecipitated with the TCR after T cell activation. Using ZAP 70-deficient T cells expressing a hybrid molecule between the SH2 domain of ZAP 70 and NEMO/IKKgamma, we showed that targeting NEMO to the immunological synapse, and more specifically its 120 N-terminal amino acids, was sufficient to selectively restore NF-kappaB activation in response to TCR ligation. Finally, we demonstrated that targeting of NEMO to the membrane of T cells was sufficient to induce constitutive NF-kappaB activation. This study shows that the localization of NEMO to the immunological synapse is important for TCR-induced NF-kappaB activation and offers a powerful system to dissect the NF-kappaB cascade in T cells. PMID- 12530973 TI - Syndecan captures, protects, and transmits HIV to T lymphocytes. AB - This study demonstrates that syndecan functions as an in trans HIV receptor. We show that syndecan, when expressed in nonpermissive cells, becomes the major mediator for HIV adsorption. This adsorption is mediated by the binding of gp120 to the heparan sulfate chains of syndecan. Although syndecan does not substitute for HIV entry receptors, it enhances the in trans infectivity of a broad range of primate lentiviruses including primary viruses produced from PBMCs. Furthermore, syndecan preserves virus infectivity for a week, whereas unbound virus loses its infectivity in less than a day. Moreover, we obtain evidence suggesting that the vast syndecan-rich endothelial lining of the vasculature can provide a microenvironment which boosts HIV replication in T cells. PMID- 12530974 TI - Impairment of NK cell function by NKG2D modulation in NOD mice. AB - Nonobese diabetic (NOD) mice, a model of insulin-dependent diabetes mellitus, have a defect in natural killer (NK) cell-mediated functions. Here we show impairment in an activating receptor, NKG2D, in NOD NK cells. While resting NK cells from C57BL/6 and NOD mice expressed equivalent levels of NKG2D, upon activation NOD NK cells but not C57BL/6 NK cells expressed NKG2D ligands, which resulted in downmodulation of the receptor. NKG2D-dependent cytotoxicity and cytokine production were decreased because of receptor modulation, accounting for the dysfunction. Modulation of NKG2D was mostly dependent on the YxxM motif of DAP10, the NKG2D-associated adaptor that activates phosphoinositide 3 kinase. These results suggest that NK cells may be desensitized by exposure to NKG2D ligands. PMID- 12530975 TI - A structural basis for the selection of dominant alphabeta T cell receptors in antiviral immunity. AB - We have examined the basis for immunodominant or "public" TCR usage in an antiviral CTL response. Residues encoded by each of the highly selected genetic elements of an immunodominant clonotype recognizing Epstein-Barr virus were critical to the antigen specificity of the receptor. Upon recognizing antigen, the immunodominant TCR undergoes extensive conformational changes in the complementarity determining regions (CDRs), including the disruption of the canonical structures of the germline-encoded CDR1alpha and CDR2alpha loops to produce an enhanced fit with the HLA-peptide complex. TCR ligation induces conformational changes in the TCRalpha constant domain thought to form part of the docking site for CD3epsilon. These findings indicate that TCR immunodominance is associated with structural properties conferring receptor specificity and suggest a novel structural link between TCR ligation and intracellular signaling. PMID- 12530976 TI - Extrachromosomal recombination substrates recapitulate beyond 12/23 restricted VDJ recombination in nonlymphoid cells. AB - V(D)J recombination occurs efficiently only between gene segments flanked by recombination signals (RSs) containing 12 and 23 base pair spacers (the 12/23 rule). A further limitation "beyond the 12/23 rule" (B12/23) exists at the TCRbeta locus and ensures Dbeta usage. Herein, we show that extrachromosomal V(D)J recombination substrates recapitulate B12/23 restriction in nonlymphoid cells. We further demonstrate that the Vbeta coding flank, the 12-RS heptamer/nonamer, and the 23-RS spacer each can significantly influence B12/23 restriction. Finally, purified core RAG1 and RAG2 proteins (together with HMG2) also reproduce B12/23 restriction in a cell-free system. Our findings indicate that B12/23 restriction of V(D)J recombination is cemented at the level of interactions between the RAG proteins and TCRbeta RS sequences. PMID- 12530977 TI - Dramatically increased rearrangement and peripheral representation of Vbeta14 driven by the 3'Dbeta1 recombination signal sequence. AB - V(D)J recombination is targeted by short recombination signal (RS) sequences that are relatively conserved but exhibit natural sequence variations. To evaluate the potential of RS sequence variations to determine the primary and peripheral TCRbeta repertoire, we generated mice containing specific replacement of the endogenous Vbeta14 RS with the 3'Dbeta1 RS (Vbeta14/3'DbetaRS). These mice exhibited a dramatic increase in Vbeta14(+) thymocyte numbers at the expense of thymocytes expressing other Vbetas. In addition, the percentage of peripheral Vbeta14(+) alphabeta T lymphocytes was similarly increased. Strikingly, this altered Vbeta repertoire resulted predominantly from a higher relative level of primary Vbeta14/3'DbetaRS rearrangement to DbetaJbeta complexes, despite the ability of the 3'Dbeta1 RS to break B12/23 restriction and allow direct rearrangement of Vbeta14/3'DbetaRS to Jbeta segments. PMID- 12530978 TI - The soluble form of IL-1 receptor accessory protein enhances the ability of soluble type II IL-1 receptor to inhibit IL-1 action. AB - Regulation of the activity of the proinflammatory cytokine IL-1 is complex, involving transcriptional and translational control, precursor processing, a receptor antagonist (IL-1ra), and a decoy receptor. Here we report that the soluble form of the IL-1 receptor accessory protein (AcP) increases the affinity of binding of human IL-1alpha and IL-1beta to the soluble human type II IL-1 receptor by approximately 100-fold, while leaving unaltered the low binding affinity of IL-1ra. Soluble AcP is present in normal human serum at an average concentration greater than 300 ng/ml. These findings suggest that the soluble form of IL-1R AcP contributes to the antagonism of IL-1 action by the type II decoy receptor, adding another layer of complexity to the regulation of IL-1 action. PMID- 12530979 TI - Peptide diffusion, protection, and degradation in nuclear and cytoplasmic compartments before antigen presentation by MHC class I. AB - Antigenic peptides generated by the proteasome have to survive a peptidase containing environment for presentation by MHC class I molecules. We have visualized the fate and dynamics of intracellular peptides in living cells. We show that peptides are distributed over two different but interconnected compartments, the cytoplasm and the nucleus, and diffuse rapidly through and between these compartments. Since TAP is excluded from the nuclear face of the nuclear envelope, nuclear peptides have to leave the nucleus to contact TAP. Thereby, these peptides encounter cytosolic peptidases that degrade peptides within seconds unless bound to chromatin. Since peptide degradation is far more efficient than translocation, many peptides will be lost for antigen presentation by MHC class I molecules. PMID- 12530980 TI - Intrinsic requirement for zinc finger transcription factor Gfi-1 in neutrophil differentiation. AB - We report essential roles of zinc finger transcription factor Gfi-1 in myeloid development. Gene-targeted Gfi-1(-/-) mice lack normal neutrophils and are highly susceptible to abscess formation by gram-positive bacteria. Arrested, morphologically atypical, Gr1(+)Mac1(+) myeloid cells expand with age in the bone marrow. RNAs encoding primary but not secondary or tertiary neutrophil (granulocyte) granule proteins are expressed. The atypical Gr1(+)Mac1(+) cell population shares characteristics of both the neutrophil and macrophage lineages and exhibits phagocytosis and respiratory burst activity. Reexpression of Gfi-1 in sorted Gfi-1(-/-) progenitors ex vivo rescues neutrophil differentiation in response to G-CSF. Thus, Gfi-1 not only promotes differentiation of neutrophils but also antagonizes traits of the alternate monocyte/macrophage program. PMID- 12530981 TI - Virus subversion of the MHC class I peptide-loading complex. AB - Many viral proteins modulate class I expression, yet, in general, their mechanisms of specific class I recognition are poorly understood. The mK3 protein of gamma(2)-Herpesvirus 68 targets the degradation of nascent class I molecules via the ubiquitin/proteasome pathway. Here, we identify cellular components of the MHC class I assembly machinery, TAP and tapasin, that are required for mK3 function. mK3 failed to regulate class I in TAP- or tapasin-deficient cells, and mK3 interacted with TAP/tapasin, even in the absence of class I. Expression of mK3 resulted in the ubiquitination of TAP/tapasin-associated class I, and mutants of class I incapable of TAP/tapasin interaction were unaffected by mK3. Thus, mK3 subverts TAP/tapasin to specifically target class I molecules for destruction. PMID- 12530982 TI - Neonates support lymphopenia-induced proliferation. AB - T cells expand without intentional antigen stimulation when transferred into adult lymphopenic environments. In this study, we show that the physiologic lymphopenic environment existing in neonatal mice also supports CD4 T cell proliferation. Strikingly, naive CD4 T cells that proliferate within neonates acquire the phenotypic and functional characteristics of memory cells. Such proliferation is inhibited by the presence of both memory and naive CD4 T cells, is enhanced by 3-day thymectomy, is independent of IL-7, and requires a class II MHC-TCR interaction and a CD28-mediated signal. CD44(bright) CD4 T cells in neonates have a wide repertoire as judged by the distribution of Vbeta expression. Thus, lymphopenia-induced T cell proliferation is a physiologic process that occurs during the early postnatal period. PMID- 12530983 TI - The Wiskott-Aldrich syndrome protein acts downstream of CD2 and the CD2AP and PSTPIP1 adaptors to promote formation of the immunological synapse. AB - The Wiskott-Aldrich syndrome protein (WASp) couples actin cytoskeletal rearrangement to T cell activation, but the mechanisms involved are unknown. Here, we show that antigen-induced formation of T cell:APC conjugates and synapses is abrogated in WASp-deficient T cells and that CD2 engagement evokes interactions between the proline-rich region required for WASp translocation to the synapse and the PSTPIP1 adaptor SH3 domain and between the PSTPIp1 coiled coil domain and both CD2 and another CD2-binding adaptor, CD2AP. The induced colocalization of these proteins at the synapse is disrupted by expression of coiled-coil domain-deleted PSTPIP1. These data, together with the impairment in CD2-induced actin polymerization observed in WASp-deficient cells, suggest that PSTPIP1 acts downstream of CD2/CD2AP to link CD2 engagement to the WASp-evoked actin polymerization required for synapse formation and T cell activation. PMID- 12530985 TI - Lessons from the blind. PMID- 12530984 TI - Antigen-specific suppression of a primed immune response by dendritic cells mediated by regulatory T cells secreting interleukin-10. AB - Antigen-specific suppression of a previously primed immune response is a major challenge for immunotherapy of autoimmune disease. RelB activation is required for myeloid DC differentiation. Here, we show that antigen-exposed DCs in which RelB function is inhibited lack cell surface CD40, prevent priming of immunity, and suppress previously primed immune responses. DCs generated from CD40 deficient mice similarly confer suppression. Regulatory CD4+ T cells induced by the DCs transfer antigen-specific "infectious" tolerance to primed recipients in an interleukin-10-dependent fashion. Thus CD40, regulated by RelB activity, determines the consequences of antigen presentation by myeloid DCs. These observations have significance for autoimmune immunotherapy and suggest a mechanism by which peripheral tolerance might be constitutively maintained by RelB(-) CD40(-) DCs. PMID- 12530986 TI - Circadian rhythm sleep disorders: lessons from the blind. AB - As totally blind people cannot perceive the light-dark cycle (the major synchroniser of the circadian pacemaker) their circadian rhythms often "free run" on a cycle slightly longer than 24 h. When the free-running sleep propensity rhythm passes out of phase with the desired time for sleep, night-time insomnia and daytime sleepiness result. It has recently been shown that daily melatonin administration can entrain the circadian pacemaker, thereby correcting this burdensome circadian sleep disorder. The primary purpose of this review is to elevate awareness of circadian sleep disorders in totally blind people (especially free-running rhythms) and to provide some guidance for clinical management. An additional goal is to show how research on sleep and circadian rhythms in the totally blind can contribute insights into the scientific understanding of the human circadian system. 2001 Harcourt Publishers Ltd PMID- 12530987 TI - Haematological aspects of obstructive sleep apnoea. AB - Obstructive sleep apnoea (OSA) is associated with increased cardiovascular morbidity and mortality. Several studies indicate an influence of OSA on haematological features. There is evidence of elevated platelet activation and increased haematocrit in OSA. Studies investigating the underlying cause of polycythaemia in OSA reveal different results: increased haematocrit in OSA might be due to enhanced erythropoietin excretion and/or plasma volume dysregulation based on altered release of volume-regulating hormones such as atrial natriuretic peptide (ANP). This review addresses haematological issues associated with OSA, especially platelet function, haematocrit and haemoconcentration. 2001 Harcourt Publishers Ltd PMID- 12530988 TI - Outcome measurement in sleep medicine practice and research. Part 2: assessment of neurobehavioral performance and mood. AB - Neurobehavioral performance and alterations in mood consequent to sleep disorders and their treatment has recently been the focus of clinical investigations. Primarily in patients with obstructive sleep apnea (OSA), there has been increased interest in the effects of sleep fragmentation and hypoxemia on the brain's ability to process information and to alter affect. The purpose of the second part of this two-part series is to describe measures that could be applied to document the impact of sleep disorders on neurobehavioral performance and mood, discuss factors affecting the selection of measures for research and practice, and to describe evidence generated by the use of these instruments in research. The neurobehavioral deficits that accompany sleep deprivation can be categorised as decrements in cognitive throughput, working memory and sustained attention. Usually evaluated using tasks of short duration, impairments associated with OSA have included impaired information processing and decline in the total number of completed and/or correct responses per unit time. Using assessments of working memory, including short recall maneuvers involving words or paragraphs, investigators have documented in sleep apnea patients the inability to consolidate and recall material producing deficits in the recollection and retention of new information. Evaluations of sustained attention appraise reaction time, the ability to remain on task, and the number of errors of omission and false responses. Evidence suggests that OSA patients display slowing of response time as well as increased errors, lapses and number of false responses. Similar deficits have been documented with sustained and divided attention tasks that present the respondent with challenges in tracking and reaction response, tasks required for driving. Although untreated sleep apnea patients were more impaired than normal controls, their performance on such tasks was greatly enhanced following CPAP treatment. In addition to substantial clinically meaningful improvements in many areas of neurobehavioral performance, changes in mood have also accompanied treatment for sleep apnea. 2001 Harcourt Publishers Ltd PMID- 12530989 TI - Sleep EEG, depression and gender. AB - Despite clear evidence of an intimate connection between sleep, major depressive disorders (MDD) and gender, few studies have explored gender differences in either sleep or in wakefulness in patients as a means of better understanding the psychogenic components of MDD. Indeed, few sleep studies focus on characterising gender differences in any population. This paper will present a review of the literature on gender differences in sleep and depression. The theoretical and clinical implications of the findings will also be discussed. The premise of the present review is that there is an inherent increased vulnerability to depression in women that arises out of basic gender differences in brain organisation and state regulation, particularly in response to a "biological challenge" during sleep. It is argued that the inherent properties of organisation and regulation of sleep EEG in healthy men and women, elicited under challenge conditions, show gender-specific vulnerability to organisational abnormalities that model homeostatic abnormalities in depressed men and women and contribute to the genesis of depression. 2001 Harcourt Publishers Ltd PMID- 12530990 TI - The process of falling asleep. AB - The process of falling asleep can best be measured by considering a convergence of behavioural, EEG, physiological and subjective information. Doing so allows one to see sleep processes as they unfold, but relying on any single sleep index can bias the description of this complex process. The studies reviewed do not support the idea that sleep begins "in a moment", but rather that entry into sleep is a continuous, interwoven series of changes which begin in relaxed drowsiness and continue through stage 1, often into the first minutes of stage 2. The transition from waking brain to sleeping brain is traced accurately by Hori's nine-stage EEG system. Event-related potential (ERP) studies map complex changes in information processing as sleep begins, while quantitative EEG investigations have identified important spatiotemporal re-organisations of primary EEG frequencies which take place as one moves from waking to sleeping mode. To consider evidence from multiple levels of analysis, a three step electrophysiological model of central nervous system (CNS) regulation during sleep onset is proposed: initial processes appear to be alpha-related; intermediate processes, poorly studied to date, parallel the development of theta and vertex sharp wave activity, while the processes which terminate wakefulness are sigma sleep spindle-related. Clinical investigations of the sleep onset period in people with narcolepsy, insomnia, depression or sleep apnoea appear to indicate the presence of relatively unique electrophysiological signatures which may be of clinical significance. 2001 Harcourt Publishers Ltd PMID- 12530991 TI - Strange bedfellows? PMID- 12530992 TI - Iron, brain and restless legs syndrome. AB - Iron is the most important transitional metal in the body, as it is implicated in many metabolic processes, mostly related to its capacity as an electron donor/acceptor. Iron deficiency has been long been known to cause anaemia, iron excess to cause haemochromatosis. As excess free iron can cause oxidative damage, it is important that the levels of iron in the body are tightly regulated which appears to be done only by digestive absorption, as there is no known regulating mechanism for elimination of iron. The amount of free iron is also kept to a minimum thanks to binding to transferrin for transport, and to ferritin for storage. Recent research has put emphasis on the possible role of excess iron in the brain in several degenerative diseases. Iron deficiency in the central nervous system is known to cause motor impairment and cognitive deficits; more recently, it has been suggested that it may play a role in the pathophysiology of the restless leg syndrome. 2001 Harcourt Publishers Ltd PMID- 12530993 TI - Sleep, sleepiness, sleep disorders and alcohol use and abuse. AB - The study of ethanol's effects on sleep has a long history dating back to the work of Nathaniel Kleitman. This paper reviews the extensive literature describing ethanol's effects on the sleep of healthy normals and alcoholics and the newer literature that describes its interactive effects on daytime sleepiness, physiological functions during sleep, and sleep disorders. Ethanol initially improves sleep in non-alcoholics at both low and high doses with disturbance in the second half of the night sleep at high doses. Tolerance develops to the initial beneficial effects. In alcoholics sleep is disturbed both while drinking and for months of abstinence and the nature of the abstinent sleep disturbance is predictive of relapse. Ethanol interacts to exacerbate daytime sleepiness and sleep-disordered breathing, even inducing apnea in persons at risk. Ethanol's effects on other physiological functions during sleep and other sleep disorders has yet to be documented. 2001 Harcourt Publishers Ltd PMID- 12530994 TI - Yawning. AB - YAWNING IS A COMMON PHYSIOLOGICAL EVENT THAT CAN BE DIVIDED INTO THREE DISTINCT PHASES: a long inspiratory phase, a brief acme and a rapid expiration. The aim of yawning is not yet well defined. However this semi-voluntary event increases vigilance and aims to alert when drowsiness occurs. Yawning probably has an important role for social communication as well. Yawning can be responsible for pain, luxation or even transient ischaemic attack. Abnormal yawning is present in various pathologies: migraine, Parkinson's disease, tumours, psychiatric diseases, infections or iatrogenic pathologies. The neuro-pharmacology of yawning is complex and knowledge of its mechanisms is incomplete. While under the control of several neurotransmitters, yawning is largely affected by dopamine. Dopamine may activate oxytocin production in the paraventricular nucleus of the hypothalamus. Oxytocin may then activate cholinergic transmission in the hippocampus and, finally, acetylcholine might induce yawning via the muscarinic receptors of the effectors. This is an over-simplification; many other molecules can modulate yawning, such as nitric oxide, glutamate, GABA, serotonin, ACTH, MSH, sexual hormones and opium derivate peptides. Dopamine involvement in yawning could have practical applications in the study of new drugs or the exploration of neurological diseases such as migraine or psychosis. 2001 Harcourt Publishers Ltd PMID- 12530995 TI - Agrypnia excitata: clinical features and pathophysiological implications. AB - Fatal familial insomnia, Morvan's chorea and delirium tremens share the same clinical features: severe insomnia and mental confusion with dream enactment, associated with motor and autonomic activation. Polygraphically, they share an inability to generate slow wave sleep. Agrypnia excitata is the term which aptly defines this peculiar medical condition. In fatal familial insomnia, the syndrome is due to a functional imbalance between activating and deactivating structures within the limbic system provoked by the atrophy of the mediodorsal and anteroventral thalamic nuclei. In Morvan's chorea and delirium tremens, a functional imbalance within the thalamolimbic circuits might be explained by the accumulation of some antireceptor antibodies and by a transient prevalence of excitatory over inhibitory synapses, down-regulated by chronic alcohol abuse, respectively. The selective disappearance of slow sleep (i.e. sleep spindles and delta rhythms) characterizing the agrypnia excitata syndrome, together with other clinical and experimental findings, suggests that sleep can be divided into three types. The most archaic form of sleep corresponding to stage 1 non-REM sleep is shared by man and poikilothermic animals and generated within activating and deactivating neuronal poles located in the basal forebrain, hypothalamus and brain stem; the other two forms of sleep, slow wave sleep and paradoxical sleep, confined to homeothermic animals, are generated in the thalamus and pontine reticular formation respectively. 2001 Harcourt Publishers Ltd PMID- 12530997 TI - Idiopathic hypersomnia: a neurological dilemma. PMID- 12530996 TI - Brain structures and mechanisms involved in the generation of NREM sleep: focus on the preoptic hypothalamus. AB - Four lines of research have greatly increased our understanding of the hypothalamic preoptic area (POA) sleep-promoting system. First, sleep-active neurons within the POA have been identified using both electrophysiological recording and immediate early gene protein (c-Fos) staining methods. Segregated sleep-active neurons were found in ventrolateral and median POA (VLPO and MnPN). Additional sleep-active neurons may be intermixed with non-sleep specific neurons in other POA regions and the adjacent basal forebrain. Second, the putative sleep factors, adenosine and prostaglandin D2, were found to excite sleep-active neurons. Other sleep factors may also modulate these sleep-active populations. Third, many sleep-active neurons are warm-sensitive neurons (WSNs). WSNs are identified by excitatory responses to small increases in local POA temperature. The same local POA thermal stimuli strongly modulate sleep propensity and EEG delta activity within sleep. Interactions between sleep regulation and thermoregulation are consistent with studies of circadian sleep propensity, prolonged sleep deprivation in rats, and species differences in sleep amounts. Fourth, sleep-active neurons were found to co-localize the inhibitory neurotransmitter, gamma-aminobutyric acid and to have projections to arousal related neuronal subgroups in the posterior hypothalamus and midbrain. Sleep active and arousal-related neurons exhibit reciprocal changes in discharge across the wake-NREM-REM cycle, and activation of WSNs suppresses the neuronal activity of some arousal-related neuronal groups. These studies establish mechanisms by which POA hypnogenic neurons can inhibit EEG and behavioral arousal. In addition, there is evidence that arousal-related neurotransmitters inhibit VLPO sleep active neurons. Mutually inhibitory interactions between sleep-promoting and the arousal system provide a substrate for a <<<>>>. 2001 Harcourt Publishers Ltd PMID- 12530998 TI - Idiopathic Hypersomnia. AB - In contrast to narcolepsy and the Kleine-Levin syndrome, idiopathic hypersomnia is a recently described sleep disorder. Absence of associated clinical features such as cataplexy or megaphagia and characteristic polysomnographic features such as sleep-onset REM episodes render positive diagnosis more uncertain in idiopathic hypersomnia than in the fwo former conditions. Consequently there has been an unfortunate tendency to label all difficult to classify cases of excessive daytime sleepiness as idiopathic hypersomnia. At present due to the description of new disorders such as upper airway resistance syndrome, narcolepsy without cataplexy, delayed sleep phase syndrome, all of which were formerly confused with idiopathic hypersomnia and the clear identification of a "polysymptomatic" or "classic" form of idiopathic hypersomnia, the limits of the disorder become more precise. Still there are a number of cases of isolated excessive daytime sleepiness with no prolonged night sleep, no difficulty waking up, which lay between narcolepsy and genuine idiopathic hypersomnia. Thus there is a definite need to further develop laboratory investigations to help identify and classify these cases. Moreover pathophysiology and pathogenesis are still in their infancy and efforts have to be pursued in this direction. Treatment has not made consistent progress except for the use of a new wake promoting compound, modafinil, which has not yet been evaluated in controlled studies. PMID- 12530999 TI - Idiopathic hypersomnia: a diagnostic dilemma. A commentary of "Idiopathic hypersomnia" (M. Billiard and Y. Dauvilliers). PMID- 12531000 TI - Beta EEG activity and insomnia. AB - To date there have been seven studies which find that beta EEG is elevated at around sleep onset and during polysomnographic sleep in patients with insomnia. These findings suggest that insomnia may be characterized by central nervous system (CNS) hyperarousal. In this article, the seven studies are critically reviewed, two theoretical perspectives on beta EEG are presented, and the concept of hyperarousal as a three component process is discussed. PMID- 12531002 TI - Do increases in beta EEG activity uniquely reflect insomnia? A commentary on "Beta EEG activity and insomnia" (M. L. Perlis et al.). PMID- 12531001 TI - Response to "Idiopathic hypersomnia: a diagnostic dilemma" (J. Montplaisir and L. Fantini). PMID- 12531003 TI - Response to "Do increases in beta EEG activity uniquely reflect insomnia?" (C. H. Bastein and M. H. Bonnet). AB - We appreciate the thoughtful comments by Drs, Bastien and Bonnet regarding our article and the opportunity provided by the Editors to respond to the issues raised. The reviewers make several critical points concerning both the beta EEG in insomnia literature and the neurocognitive model. Our summary of their concerns and our thoughts on the issues they raise are as follows. PMID- 12531004 TI - Sleep and pain. AB - Noxious stimuli and painful disorders interfere with sleep, but disturbances in sleep also contribute to the experience of pain.Chronic paroxysmal hemicrania and possibly cluster headaches are related to REM sleep. Whereas headache is associated with snoring and sleep apnea, morning headaches are not specific for any primary sleep disorder. Nevertheless, the management of the sleep disorder ameliorates both morning headache and migraine.Noxious stimuli administered into muscles during slow-wave sleep (SWS) result in decreases in delta and sigma but an increase in alpha and beta EEG frequencies during sleep. Noise stimuli that disrupt SWS result in unrefreshing sleep, diffuse musculoskeletal pain, tenderness, and fatigue in normal healthy subjects. Such symptoms accompany alpha EEG sleep patterns that often occur in patients with fibromyalgia. The alpha EEG patterns include phasic and tonic alpha EEG sleep as well as periodic K alpha EEG sleep or frequent periodic cyclical alternating pattern. Moreover, alpha EEG sleep, as well as sleep-related breathing disorder and periodic limb movement disorder, occur in some patients with fibromyalgia, rheumatoid arthritis and osteoarthritis. Depression and not alpha EEG sleep are features of somatoform pain disorder. Disturbances in sleep, pain behaviour and psychological distress influence return to work in workers who have suffered a soft tissue injury, e.g. low back pain. Patients with irritable bowel disorder have disturbed sleep and have increased REM sleep. In conclusion, there is a reciprocal relationship between sleep quality and pain. The recognition of disturbed or unrefreshing sleep influences the management of painful medical disorders. PMID- 12531005 TI - The search for the molecular correlates of sleep and wakefulness. AB - Knowledge of the molecular correlates of sleep and wakefulness is essential if we are to understand the restorative processes occurring during sleep and the cellular mechanisms underlying sleep regulation. In order to determine what molecular changes occur during the sleep-waking cycle, we have recently performed a systematic screening of gene expression in the brain of sleeping, sleep deprived and spontaneously awake rats. Out of the approximately 10 000 genes screened so far, a small minority ( approximately 0.5%) was differentially expressed in the cerebral cortex across behavioral states. Most genes were upregulated in wakefulness and sleep deprivation relative to sleep, while only a few had higher expression in sleep relative to wakefulness and sleep deprivation. Almost all the genes upregulated in sleep, and several genes upregulated in wakefulness and sleep deprivation, did not match any known sequence. Known genes that were upregulated in wakefulness and sleep deprivation could be grouped into functional categories: immediate early genes/transcription factors, genes related to energy metabolism, growth factors/adhesion molecules, chaperones/heat shock proteins, vesicle- and synapse-related genes, neurotransmitter/hormone receptors, neurotransmitter transporters, enzymes, and others. Although the characterization of the molecular correlates of sleep, wakefulness and sleep deprivation is still in progress, it is already apparent that the transition from sleep to waking can affect basic cellular functions such as RNA and protein synthesis, neural plasticity, neurotransmission, and metabolism. PMID- 12531006 TI - Yawning. PMID- 12531008 TI - DNA structure dependent checkpoints as regulators of DNA repair. AB - Checkpoint proteins were initially identified because their loss of function resulted in defects in cell cycle arrest in response to genotoxic treatments. Initially, the analysis of checkpoint pathways concentrated on their function as signal transducers and how the checkpoint signals were communicated to the core cell cycle machinery and transcriptional apparatus. Although some of the early genetic analysis indicated a complex relationship between DNA replication, DNA repair and the checkpoint pathways, it is only now becoming apparent that checkpoint proteins regulate multiple DNA repair and replication functions. Furthermore, recent data suggest that some checkpoint proteins may participate directly in DNA repair events. In this review I summarise the current models for DNA structure-dependent checkpoint activation and review the evidence linking checkpoint proteins both directly and indirectly to DNA repair. PMID- 12531009 TI - A:T --> G:C base pair substitutions occur at a higher rate than other substitution events in Pms2 deficient mouse cells. AB - The mismatch repair pathway involves multiple proteins that are required to correct DNA polymerase generated mismatches before they become mutations. It has been shown recently, that the predominant base-pair substitution events leading to loss of endogenous Aprt activity in Pms2 null mouse cells are A:T --> G:C mutations (Oncogene 21 (2002) 1768, Oncogene 21 (2002) 2840). To determine if this observation could be explained by an increased rate of A:T --> G:C mutations relative to other base-pair substitutions, we developed a reversion assay to examine G:C --> A:T, C:G --> A:T, and A:T --> G:C mutations within mouse Aprt in a Pms2 null mouse kidney cell line. The results demonstrated a 6-50-fold increase in the rate of the A:T --> G:C mutations relative to the other base-pair substitutions. Additional work demonstrated that growth of the Pms2 null cells in antioxidant containing medium reduced the rate of the A:T --> G:C mutations. The results are discussed with regards to the role of mismatch repair proteins in preventing base-pair substitutions, including those induced by oxidative stress. PMID- 12531010 TI - Frameshifts and deletions during in vitro translesion synthesis past Pt-DNA adducts by DNA polymerases beta and eta. AB - DNA polymerases beta (pol beta ) and eta (pol eta ) are the only two eukaryotic polymerases known to efficiently bypass cisplatin and oxaliplatin adducts in vitro. Frameshift errors are an important aspect of mutagenesis. We have compared the types of frameshifts that occur during translesion synthesis past cisplatin and oxaliplatin adducts in vitro by pol beta and pol eta on a template containing multiple runs of nucleotides flanking a single platinum-GG adduct. Translesion synthesis past platinum adducts by pol beta resulted in approximately 50% replication products containing single-base deletions. For both adducts the majority of -1 frameshifts occurred in a TTT sequence 3-5 bp upstream of the DNA lesion. For pol eta, all of the bypass products for both cisplatin and oxaliplatin adducts contained -1 frameshifts in the upstream TTT sequence and most of the products of replication on oxaliplatin-damaged templates had multiple replication errors, both frameshifts and misinsertions. In addition, on platinated templates both polymerases generated replication products 4-8 bp shorter than the full-length products. The majority of short cisplatin-induced products contained an internal deletion which included the adduct. In contrast, the majority of oxaliplatin-induced short products contained a 3' terminal deletion. The implications of these in vitro results for in vivo mutagenesis are discussed. PMID- 12531011 TI - The embryonic lethality in DNA ligase IV-deficient mice is rescued by deletion of Ku: implications for unifying the heterogeneous phenotypes of NHEJ mutants. AB - There are two general pathways by which multicellular eukaryotes repair double strand DNA breaks (DSB): homologous recombination (HR) and nonhomologous DNA end joining (NHEJ). All mammalian mutants in the NHEJ pathway demonstrate a lack of B and T lymphocytes and ionizing radiation sensitivity. Among these NHEJ mutants, the DNA-PK(cs) and Artemis mutants are the least severe, having no obvious phenotype other than the general defects described above. Ku mutants have an intermediate severity with accelerated senescence. The XRCC4 and DNA ligase IV mutants are the most severe, resulting in embryonic lethality. Here we show that the lethality of DNA ligase IV-deficiency in the mouse can be rescued when Ku86 is also absent. To explain the fact that simultaneous gene mutations in the NHEJ pathway can lead to viability when a single mutant is not viable, we propose a nuclease/ligase model. In this model, disrupted NHEJ is more severe if the Artemis:DNA-PK(cs) nuclease is present in the absence of a ligase, and Ku mutants are of intermediate severity, because the nuclease is less efficient. This model is also consistent with the order of severity in organismal phenotypes; consistent with chromosomal breakage observations reported here; and consistent with the NHEJ mutation identified in radiation sensitive human SCID patients. PMID- 12531012 TI - Anti-tumour compounds illudin S and Irofulven induce DNA lesions ignored by global repair and exclusively processed by transcription- and replication-coupled repair pathways. AB - Illudin S is a natural sesquiterpene drug with strong anti-tumour activity. Inside cells, unstable active metabolites of illudin cause the formation of as yet poorly characterised DNA lesions. In order to identify factors involved in their repair, we have performed a detailed genetic survey of repair-defective mutants for responses to the drug. We show that 90% of illudin's lethal effects in human fibroblasts can be prevented by an active nucleotide excision repair (NER) system. Core NER enzymes XPA, XPF, XPG, and TFIIH are essential for recovery. However, the presence of global NER initiators XPC, HR23A/HR23B and XPE is not required, whereas survival, repair and recovery from transcription inhibition critically depend on CSA, CSB and UVS, the factors specific for transcription-coupled NER. Base excision repair and non-homologous end-joining of DNA breaks do not play a major role in the processing of illudin lesions. However, active RAD18 is required for optimal cell survival, indicating that the lesions also block replication forks, eliciting post-replication-repair-like responses. However, the translesion-polymerase DNA pol eta is not involved. We conclude that illudin-induced lesions are exceptional in that they appear to be ignored by all of the known global repair systems, and can only be repaired when trapped in stalled replication or transcription complexes. We show that the semisynthetic illudin derivative hydroxymethylacylfulvene (HMAF, Irofulven), currently under clinical trial for anti-tumour therapy, acts via the same mechanism. PMID- 12531013 TI - Sorting the consequences of ionizing radiation: processing of 8-oxoguanine/abasic site lesions. AB - Clustered DNA damage is a hallmark of ionizing radiation. These complex lesions, composed of any combination of oxidized bases, abasic sites, or strand breaks within one helical turn, create a tremendous challenge for the base excision repair system, which must process the damage without generating cytotoxic double strand breaks (DSB). Clustered lesions affect the DNA incision activity of DNA glycosylases and AP endonucleases. Different levels of enzyme inhibition are dependent on lesion identity, orientation and separation. Very little is known about the simultaneous action of both classes of enzymes, which may lead to the creation of DSB. We have developed a novel substrate system of double-labeled hairpin duplexes, which allows the simultaneous determination of enzyme incision and formation of DBS. We use this system to study the processing of four clustered 8-oxoguanine/abasic site lesions by purified mouse Ogg1, human Ape1 and mouse embryonic stem cell nuclear extracts. Ape1 activity is least affected by the presence of a nearby oxidized base. In contrast, an abasic site inhibits the glycosylase and lyase activities of Ogg1 in an orientation-dependent manner. The combined action of both enzymes leads to the preferential formation of DSB with 5'-overhang ends. Processing of clusters by nuclear extracts displayed similar patter of enzyme inhibition and the same preference for avoiding double strand breaks with 3'-overhang ends. PMID- 12531014 TI - Cytotoxicity and mutagenesis induced by singlet oxygen in wild type and DNA repair deficient Escherichia coli strains. AB - Singlet oxygen ((1)O(2)) is a product of several biological processes and can be generated in photodynamic therapy, through a photosensitization type II mechanism. (1)O(2) is able to interact with lipids, proteins and DNA, leading to cell killing and mutagenesis, and can be directly involved with degenerative processes such as cancer and aging. In this work, we analyzed the cytotoxicity and mutagenesis induced after direct treatment of wild type and the DNA repair fpg and/or mutY deficient Escherichia coli strains with disodium 3,3'-(1,4 naphthylidene) diproprionate endoperoxide (NDPO(2)), which releases (1)O(2) by thermodissociation. The treatment induced cell killing and mutagenesis in all strains, but the mutY strain showed to be more sensitive. These results indicate that even (1)O(2) generated outside bacterial cells may lead to DNA damage that could be repaired by pathways that employ MutY protein. As (1)O(2) is highly reactive, its interaction with cell membranes may generate secondary products that could react with DNA, leading to mutagenic lesions. PMID- 12531015 TI - A new damage limitation exercise: ironing (Fe(II)) out minor DNA methylation lesions. PMID- 12531016 TI - Structural and functional conservation of error-free DNA postreplication repair in Schizosaccharomyces pombe. AB - DNA postreplication repair (PRR) is a cellular process by which cells survive replication-blocking lesions without removing the lesion. In the budding yeast Saccharomyces cerevisiae, MMS2 plays a key role in the error-free PRR pathway: the mms2 null mutant displays an increased spontaneous mutation rate and sensitivity to a variety of DNA damaging agents. In contrast, its human homologs appear to play a different role. In order to address whether the MMS2-mediated PRR pathway is conserved in eukaryotes, we isolated a Schizosaccharomyces pombe cDNA homologous to MMS2, which we named spm2(+). Using spm2(+) as a bait in a yeast two-hybrid screen, we identified a fission yeast cDNA homologous to UBC13 from various species and named it spu13(+). Two-hybrid analysis confirmed physical interaction between Spm2 and Spu13, and between Spm2 and budding yeast Ubc13. Genetic analysis shows that both spm2(+) and spu13(+) are able to functionally complement the corresponding budding yeast mutants. Furthermore, deletion of either spm2(+), spu13(+) or both genes from fission yeast results in an increased sensitivity to DNA damaging agents, suggesting that spm2(+) and spu13(+) indeed function in PRR. The fact that the spm2(-) spu13(-) double mutant showed sensitivity similar to that of the single mutant indicates that these two gene products act at the same step. Hence, our data strongly support the hypothesis that the PRR function mediated by UBC13-MMS2 is conserved throughout eukaryotes. PMID- 12531017 TI - Mutational specificity of mice defective in the MTH1 and/or the MSH2 genes. AB - Oxidative damage of nucleotides within DNA or precursor pools caused by oxygen radicals is thought to play an important role in spontaneous mutagenesis, as well as carcinogenesis and aging. In particular, 8-oxodGTP and 2-OHdATP are potent mutagenic substrate for DNA synthesis. Mammalian MTH1 catalyzes hydrolysis of these mutagenic substrates, suggesting that it functions to prevent mutagenesis caused by these oxidized nucleotides. We have established MTH1(-/-) mice lacking the 8-oxodGTPase activity, which were shown to be susceptible to lung, liver and stomach cancers. To examine in vivo mutation events due to the MTH1-deficiency, a reporter gene, rpsL of Escherichia coli, was introduced into MTH1(-/-) mice. Interestingly, the net frequency of rpsL(-) forward mutants showed no apparent increase in MTH1(-/-) mice as compared to MTH1(+/+) mice. However, we found differences between these two genotypes in the class- and site-distributions of the rpsL(-) mutations recovered from the mice. Unlike MutT-deficient E. coli showing 1000-fold higher frequency of A:T-->C:G transversion than the wild type cells, an increase in frequency of A:T-->C:G transversion was not evident in MTH1 nullizygous mice. Nevertheless, the frequency of single-base frameshifts at mononucleotide runs was 5.7-fold higher in spleens of MTH1(-/-) mice than in those of wild type mice. Since the elevated incidence of single-base frameshifts at mononucleotide runs is a hallmark of the defect in MSH2-dependent mismatch repair system, this weak site-specific mutator effect of MTH1(-/-) mice could be attributed to a partial sequestration of the mismatch repair function that may act to correct mispairs with the oxidized nucleotides. Consistent with this hypothesis, a significant increase in the frequency of G:C-->T:A transversions was observed with MTH1(-/-) MSH2(-/-) mice over MSH2(-/-) mice alone. These results suggest a possible involvement of multiple anti-mutagenic pathways, including the MTH1 protein and other repair system(s), in mutagenesis caused by the oxidized nucleotides. PMID- 12531018 TI - Characterization of nuclease-dependent functions of Exo1p in Saccharomyces cerevisiae. AB - Exo1p is a member of the Rad2p family of structure-specific nucleases that contain conserved N and I nuclease domains. Exo1p has been implicated in numerous DNA metabolic processes, such as recombination, double-strand break repair and DNA mismatch repair (MMR). In this report, we describe in vitro and in vivo characterization of full-length wild-type and mutant forms of Exo1p. Herein, we demonstrate that full-length yeast Exo1p possesses an intrinsic 5'-3' exonuclease activity as reported previously, but also possesses a flap-endonuclease activity. Our study indicates that Exo1p shares similar, but not identical structure function relationships to other characterized members of the Rad2p family in the N and I nuclease domains. The two exo1p mutants we examined, showed deficiencies for both double-stranded DNA (dsDNA) 5'-3' exonuclease and flap-endonuclease activities. Examining the genetic interaction of these two exo1 mutations with rad27Delta suggest that the Exo1p flap-endonuclease activity and not the dsDNA 5' 3' exonuclease is redundant to Rad27p for viability. In addition, our in vivo results also indicate that many exo1Delta phenotypes are dependent on the complete catalytic activities of Exo1p. Finally, our findings plus those of other investigators suggest that Exo1p functions both in a catalytic and a structural capacity during DNA MMR. PMID- 12531019 TI - Functional crosstalk between hOgg1 and the helicase domain of Cockayne syndrome group B protein. AB - We have previously reported that the Cockayne syndrome group B gene product (CSB) contributes to base excision repair (BER) of 8-hydroxyguanine (8-OH-Gua) and the importance of motifs V and VI of the putative helicase domains of CSB in BER of 8 OH-Gua. To further elucidate the function of CSB in BER, we investigated its role in the pathway involving human 8-OH-Gua glycosylase/apurinic lyase (hOgg1). Depletion of CSB protein with anti-CSB antibody reduced the 8-OH-Gua incision rate of wild type cell extracts but not of CSB null and motif VI mutant cell extracts, suggesting a direct contribution of CSB to the catalytic process of 8 OH-Gua incision and the importance of its motif VI in this pathway. Introduction of recombinant purified CSB partially complemented the depletion of CSB as shown by the recovery of the incision activity. This complementation could not fully recover the deficiency of the incision activity in WCE from CS-B null and mutant cell lines, suggesting that some additional factor(s) are necessary for the full activity. Electrophoretic mobility shift assays (EMSAs) showed a defect in binding of CSB null and motif VI mutant cell extracts to 8-OH-Gua-containing oligonucleotides. We detected less hOgg1 transcript and protein in the cell extracts from CS-B null and mutant cells, suggesting hOgg1 may be the missing component. Pull-down of hOgg1 by histidine-tagged CSB and co-localization of those two proteins after gamma-radiation indicated their co-existence in vivo, particularly under cellular stress. However, we did not detect any functional and physical interaction between purified CSB and hOgg1 by incision, gel shift and yeast two-hybrid assays, suggesting that even though hOgg1 and CSB might be in a common protein complex, they may not interact directly. We conclude that CSB functions in the catalysis of 8-OH-Gua BER and in the maintenance of efficient hOgg1 expression, and that motif VI of the putative helicase domain of CSB is crucial in these functions. PMID- 12531021 TI - Additive roles of XPA and MSH2 genes in UVB-induced skin tumorigenesis in mice. AB - We have made xeroderma pigmentosum group A gene (XPA)-knockout mice (XPA(-/-) mice). The XPA(-/-) mice had no detectable activity for nucleotide excision repair (NER) and showed a high incidence of UVB-induced skin tumorigenesis. We have also found that cell lines derived from skin cancers in UVB-irradiated XPA( /-) mice become tolerant to UV-irradiation and showed abnormal UV-induced cell cycle checkpoints and decreased mismatch repair (MMR) activity. These results suggested that the MMR-downregulation may help cells escape killing by UV irradiation and thus MMR-deficient clones are selected for during the tumorigenic transformation of XPA(-/-) cells. In this report, we examined whether the incidence of UVB-induced skin tumorigenesis is enhanced in XPA(-/-)MSH2(-/-), XPA(-/-) and MSH2(-/-) mice when compared with that in wild-type mice. Our results indicate that the MSH2-deficiency caused a high incidence of spontaneous and UVB-induced skin tumorigenesis and the XPA and MSH2 genes have additive roles in the UV-induced skin tumorigenesis. PMID- 12531020 TI - Mice defective in the mismatch repair gene Msh2 show increased predisposition to UVB radiation-induced skin cancer. AB - Mice defective in the mismatch repair (MMR) gene Msh2 manifest an enhanced predisposition to skin cancer associated with exposure to UVB radiation. This predisposition is further heightened if the mice are additionally defective for the nucleotide excision repair gene Xpc. To test the hypothesis that the predisposition of Msh2 mutant mice to skin cancer reflects a mutator phenotype associated with increased proliferation of skin cells following exposure to UV radiation, Msh2 mutant mice were exposed to the tumor promoter TPA. Such mice showed a robust proliferative response in the skin, but did not manifest evidence of dysplasia or neoplasia. We conclude that the predisposition of Msh2 mice to UVB radiation-induced skin cancer reflects an interaction between the processes of mismatch repair and some other excision repair mode, the exact nature of which remains to be established. PMID- 12531022 TI - Spontaneous and bleomycin-induced genomic alterations in the progeny of Drosophila treated males depends on the Msh2 status. DNA fingerprinting analysis. AB - Deficiency in DNA mismatch repair (MMR) confers instability of simple repeated sequences and increases susceptibility to cancer. Some of the MMR genes are also implicated in other repair and cellular processes related to DNA damage response. Supposedly, lack of their function can lead to a global genomic instability, besides microsatellite instability (MSI). To study the spontaneous and induced genomic instability in germ cells, related to the Msh2 status, DNA alterations in the progeny of individual crosses of Drosophila deficient in one or two copies of the Msh2 gene, were analysed by the arbitrarily primed polymerase chain reaction (AP-PCR). The results indicate that the progeny of homozygous parents for the normal Msh2 allele (+/+) presents a significantly lower frequency of genomic alterations than those from heterozygous (+/-) or mutant homozygous (-/-) parents. In addition, the DNA damage transmitted to the progeny, after the adult parental males were exposed to bleomycin, indicates that whereas the induction of mutations related to MSI depends on the lack of the Msh2 function, the induction of other mutational events may require at least one functional Msh2 allele. Thus, the results obtained with heterozygous individuals may have special relevance for cancer development since they show that a disrupted Msh2 allele is enough to generate genomic instability in germ cells, increasing the genomic damage in the progeny of heterozygous individuals. This effect is enhanced by mutagenic stress, such as occurs after bleomycin exposure. PMID- 12531023 TI - Survival and SOS induction in cisplatin-treated Escherichia coli deficient in Pol II, RecBCD and RecFOR functions. AB - Cisplatin is a potent anticancer agent forming intrastrand-crosslinks in DNA. The efficacy of cisplatin in chemotherapy can be limited by the development of tumor resistances such as elevated DNA repair or damage tolerance. In Escherichia coli, cisplatin treatment causes induction of the SOS regulon resulting in elevated levels of DNA Pol II, DNA Pol IV, DNA Pol V, the cell division inhibitor SfiA (SulA), homologous recombination (HR) and DNA repair. In this work, the roles of Pol II and HR in facilitating resistance of E. coli to cisplatin are studied. SOS induction levels were measured by beta-galactosidase assays in cisplatin-treated and untreated E. coli PQ30 that has the lacZ gene fused to the sfiA promoter. Comparative studies were carried out with derivatives of PQ30 constructed by P1 transduction that have transposon insertions in the polB gene, the recB gene blocking the RecBCD pathway of HR and genes of the RecFOR pathway of HR. Resistance of E. coli strains to cisplatin as determined by plating experiments decreased in the following order: parent PQ30 strain, polB > recO, recR, recF > recB. Both the RecBCD and RecFOR pathways of HR are important for survival when E. coli is exposed to cisplatin, because treatment of double mutants deficient in both pathways reduced colony forming ability to 37% in 6-9min in comparison to 39 120min for single mutants. Pol II and RecF appear to function in two distinct pathways to initiate replication blocked due to damage caused by cisplatin because function of Pol II was required for survival in mutants deficient in the RecFOR pathway after 2h of cisplatin treatment. In contrast, Pol II was not required for survival in recB mutants. SOS induction was delayed in RecFOR deficient mutants but occurred at high levels in the recB mutant soon after cisplatin treatment in a RecFOR-dependent way. An SfiA independent, DNA damage dependent pathway is apparently responsible for the filamentous cells observed after cisplatin or MMC treatments of these SfiA defective strains. PMID- 12531024 TI - A single 8,5'-cyclo-2'-deoxyadenosine lesion in a TATA box prevents binding of the TATA binding protein and strongly reduces transcription in vivo. AB - 8,5'-Cyclo-2'-deoxypurine (cPu) lesions result from the action of the hydroxyl radical on DNA. These lesions represent a unique class of oxidative DNA lesions in that they are repaired by the nucleotide excision repair (NER) pathway but not by base excision repair (BER) or direct repair. Previous work has shown that cyclopurines can block mammalian DNA and RNA polymerases. Thus, these lesions are of interest because of their potential role in the neurodegeneration as well as internal cancers observed in patients with xeroderma pigmentosum (XP) who lack the capacity to carry out NER. In the present work, we found that the S-isomer of 8,5'-cyclo-2'-deoxyadenosine (cA) can prevent binding of the TATA binding protein (TBP) to the TATA box from the CMV promoter. To assess the functional importance of this effect in living cells, we transfected constructs containing a single cA in the CMV TATA box into XP cells to determine the effect of the lesion on gene expression in vivo. Using this approach, we found that the lesion reduced gene expression by approximately 75%. This effect was comparable to the effect of an inactivating mutation of the TATA box in the same promoter. These findings identify an additional biological effect of cyclopurine lesions in mammalian cells, which is the ability to interfere with transcription by preventing transcription factor binding to cognate recognition sequences. In addition, the approach we used in this study represents a novel method for assessing the effects of DNA lesions in non-transcribed sequences on gene expression in living cells. PMID- 12531025 TI - Ultraviolet photobiology: its early roots and insights into DNA repair. AB - This is the first of a series of commentaries on classic papers on DNA repair that highlight the birth of this discipline. The roots go deep, and in this first commentary, I describe some of the earliest discoveries of the mechanism of absorption of UV light in cells, and its lethal and mutagenic effects. Most remarkably, a discovery of DNA repair by the use of split doses of UV light was reported in 1919. Later commentaries will take us sequentially through the early years of DNA repair. PMID- 12531026 TI - Analysis of mouse Rad54 expression and its implications for homologous recombination. AB - Homologous recombination is one of the major pathways for repair of DNA double strand breaks (DSBs). Important proteins in this pathway are Rad51 and Rad54. Rad51 forms a nucleoprotein filament on single-stranded DNA (ssDNA) that mediates pairing with and strand invasion of homologous duplex DNA with the assist of Rad54. We estimated that the nucleus of a mouse embryonic stem (ES) cells contains on average 4.7x10(5) Rad51 and 2.4x10(5) Rad54 molecules. Furthermore, we showed that the amount of Rad54 was subject to cell cycle regulation. We discuss our results with respect to two models that describe how Rad54 stimulates Rad51-mediated DNA strand invasion. The models differ in whether Rad54 functions locally or globally. In the first model, Rad54 acts in cis relative to the site of strand invasion. Rad54 coats the Rad51 nucleoprotein filament in stoichiometric amounts and binds to the target duplex DNA at the site that is homologous to the ssDNA in the Rad51 nucleoprotein filament. Subsequently, it promotes duplex DNA unwinding. In the second model, Rad54 acts in trans relative to the site of strand invasion. Rad54 binds duplex DNA distant from the site that will be unwound. Translocation of Rad54 along the duplex DNA increases superhelical stress thereby promoting duplex DNA unwinding. PMID- 12531027 TI - A nuclear 3'-5' exonuclease proofreads for the exonuclease-deficient DNA polymerase alpha. AB - DNA replication is a highly accurate process designed to duplicate the entire genome of a cell during each cell division. The accuracy of DNA replication is derived from the balance between three important components: base selectivity by the replicative DNA polymerases (pols), exonucleolytic proofreading, and post replicative mismatch repair. Previously we identified a human 3'-5' exonuclease (exoN) whose properties suggested it may function as a proofreader for the exonuclease-deficient replicative DNA pol alpha. Purified exoN has no associated pol activity and catalyzes removal of mispaired nucleotides from DNA duplexes. Consistent with previous reports, it was found that mammalian pol alpha is inefficient at extending from mispaired DNA terminals. However, in similar reactions that included exoN, there was a 4.4-15.7-fold increase in pol alpha catalyzed elongation from mispaired base pairs. In contrast, exoN did not have a dramatic impact on the ability of exonuclease-deficient variants of Klenow (K-) and T7 polymerase to catalyze extension from mispaired DNA. Continuous DNA replication catalyzed by either pol alpha or K- generated base substitutions at a frequency of 24.3x10(-4) and 38x10(-4), respectively. ExoN restored error-free DNA replication in reactions with pol alpha whereas it did not significantly improve the accuracy of K-. These results are consistent with a functional interaction between exoN and pol alpha to ensure accurate DNA replication. PMID- 12531028 TI - Ultraviolet light-induced apoptosis is associated with S-phase in primary human fibroblasts. AB - Transcription-coupled nucleotide excision repair (tcNER)-deficient human fibroblasts are extremely sensitive to the induction of apoptosis in response to low doses of ultraviolet light (UV light), but are less sensitive to the induction of apoptosis following exposure to high doses [J. Invest. Dermatol. 117 (2001) 1162]. This seemingly paradoxical observation led us to re-evaluate the relationship between UV dose and the induction of apoptosis. Here we report that the reduction in the extent of UV-induced apoptosis in tcNER-deficient strains following exposure to elevated doses of UV light does not result from impaired gene expression alone because neither inhibitors of transcription nor inhibitors of translation blocked UV-induced apoptosis. Furthermore, UV-induced apoptosis was greatly reduced by inhibiting S-phase progression with either mimosine or serum withdrawal. Importantly, DNA synthesis following UV-irradiation occurred only at doses that induced apoptosis in these cell lines and the apoptotic cells contained nascent DNA. Moreover, deregulation of G(1)- to S-phase transition by expression of human papillomavirus E7 sensitized cells to UV-induced apoptosis. Taken together these results suggest that the induction of apoptosis requires S phase progression following UV-irradiation. PMID- 12531029 TI - DNA damage-processing in E. coli: on-going protein synthesis is required for fixation of UV-induced lethality and mutation. AB - UV irradiation of E. coli produces photoproducts in the DNA genome. In consequence, some bacteria lose viability (colony-forming ability) or remain viable as mutant cells. However, the end-points of viability inactivation (lethality) or mutation are determined by cellular processes that act on the UV damaged DNA. We have investigated the in vivo time course for processes that deal with cyclobutane pyrimidine dimers (CPD) which can be specifically removed by photoreactivation (PR). At different times during post-UV incubation, samples were challenged with PR and assayed for viability or mutation. We used excision defective E. coli B/r cells and worked under yellow light to avoid background PR. During post-UV incubation (0-100min) in fully supplemented defined medium, inactivation and mutation were initially significantly reversed by PR but the extent of this reversal decreased during continued incubation defining "fixation" of lethality or mutation, respectively. In contrast, if protein synthesis was restricted during the post-UV incubation, no fixation developed. When chloramphenicol was added to inhibit protein synthesis after 30min of supplemented post-UV incubation, at a time sufficient for expression of UV induced protein(s), fixation of lethality or mutation was still annulled (no change in the effectiveness of PR developed). Lethality fixation did progress when protein synthesis was restricted and the cells were incubated in the presence of puromycin or were either clpP or clpX defective. We discuss these and related results to suggest (1) on-going protein synthesis is required in the fixation process for lethality and mutation to sustain an effective level of a hypothetical protein sensitive to ClpXP proteolysis and (2) this protein plays a critical role in the process leading to exchange between Pol III activity and alternative polymerase activities required as each cell deals with damage in template DNA. PMID- 12531030 TI - Two budding yeast RAD4 homologs in fission yeast play different roles in the repair of UV-induced DNA damage. AB - We have identified two fission yeast homologs of budding yeast Rad4 and human xeroderma pigmentosum complementation group C (XP-C) correcting protein, designated Rhp4A and Rhp4B. Here we show that the rhp4 genes encode NER factors that are required for UV-induced DNA damage repair in fission yeast. The rhp4A deficient cells but not the rhp4B-deficient cells are sensitive to UV irradiation. However, the disruption of both rhp4A and rhp4B resulted in UV sensitivity that was greater than that of the rhp4A-deficient cells, revealing that Rhp4B plays a role in DNA repair on its own. Fission yeast has two pathways to repair photolesions on DNA, namely, nucleotide excision repair (NER) and UV damaged DNA endonuclease-dependent excision repair (UVER). Studies with the NER deficient rad13 and the UVER-deficient (Delta)uvde mutants showed the two rhp4 genes are involved in NER and not UVER. Assessment of the ability of the various mutants to remove cyclobutane pyrimidine dimers (CPDs) from the rbp2 gene locus indicated that Rhp4A is involved in the preferential repair of lesions on the transcribed DNA strand and plays the major role in fission yeast NER. Rhp4B in contrast acts as an accessory protein in non-transcribed strand (NTS) repair. PMID- 12531032 TI - The intersection between the birth of molecular biology and the discovery of DNA repair. AB - Some of the physicists and biologists who contributed to early studies on the nature of the gene employed physical tools such as ionizing and ultraviolet (UV) radiation as experimental tools. The abnormalities of gene function that resulted from these perturbations led some investigators to explore the nature of DNA damage and to the (initially providential) discovery of DNA repair. However, rapid progress in this field through the efforts of molecular biologists conversant with DNA damage and repair was hampered by (among other things) a greater intellectual interest in the nature of DNA replication and the way DNA encoded the formation of proteins. PMID- 12531031 TI - Differential intracellular localization of the human and mouse endonuclease III homologs and analysis of the sorting signals. AB - The mammalian endonuclease III homolog NTH1 is a DNA glycosylase/AP lyase that recognizes oxidized pyrimidine bases. Here, we compared the intracellular localization of human and mouse NTH1 and analyzed their sorting signals by examining expression of enhanced green fluorescent protein (EGFP)-tagged NTH1 protein. Full-length hNTH1 was sorted exclusively into nuclei. Deletion analysis showed that two basic amino acid clusters, which constitute the nuclear localization signal (NLS), are essential for nuclear sorting. Moreover, disruption of the NLS by deletion or substitution of arginine residue(s) altered the localization of the protein to mitochondria. In contrast, most mNTH1 molecules were sorted into mitochondria, with a relatively small amount localized in nuclei. Deletion analysis indicated that the mitochondrial targeting sequence of mNTH1 is contained within the N-terminal 38 amino acids. Alignment of the N terminal sequence of human and mouse NTH1 showed that mNTH1 lacks a basic amino acid cluster corresponding to one of the NLS sequences found in hNTH1. Nuclear localization of mNTH1 was increased when this NLS sequence was added to mNTH1 through the addition of appropriate amino acids. The fact that transcription of the hNTH1 gene is initiated at multiple sites indicated that three isoforms of hNTH1 protein are translated using different initiation codons. However, no difference in intracellular localization was observed among three isoforms of hNTH1 with different N-terminal sequences. PMID- 12531033 TI - Promises made. promises kept. PMID- 12531034 TI - GUEST EDITORIAL: Rational pharmacotherapy for insomnia: time for a new paradigm. PMID- 12531035 TI - Hypnotic medication in the treatment of chronic insomnia: non nocere! Doesn't anyone care? AB - Chronic primary insomnia is a recurrent condition that negatively effects the daily functioning of patients, diminishing the quality of their lives. It is associated with, and in some situations, is a risk factor in both psychiatric (depression) and physical (cardiovascular) illness. Treatment effectiveness has been shown in the short term for both drug (benzodiazepine and benzodiazepine agonists) and behavioral treatment. Expert opinion has strongly advised against long-term drug treatment because of concerns about residual sedative effects, memory impairment, falls, respiratory depression, rebound insomnia, medication abuse, dose escalation, dependency and withdrawal difficulties, and an increased risk of death possibly associated with the current hypnotic medications. Many of these concerns could be made against using these agents at all. Worries about these potential problems are challenged by the widespread clinical practice of using hypnotic drugs long-term without any of these difficulties developing and with patients who feel their sleep and daily function function is improved with the nightly use of their sleeping pill. The ability to mount a randomized, placebo-controlled, parallel group, double-blind trial of hypnotic medication in primary insomnia may not be possible. We may have to develop large systematic clinical databases, a number of case series in effect, to monitor both emergent symptoms and possible clinical effectiveness. There is the additional concern that there is a reluctance to examine the long-term drug treatment of insomnia. This reluctance may reflect a negative moral judgement about treating primary insomnia with drugs, a sort of <<<>>>, rather than just a data based judiciousness. PMID- 12531036 TI - Non-benzodiazepines for the treatment of insomnia. AB - Benzodiazepine hypnotics, the mainstay of pharmacological treatment for insomnia, have been associated with altered sleep architecture, psychomotor and memory impairment, rebound insomnia, withdrawal effects, tolerance, dependence, abuse potential and respiratory depression. Non-benzodiazepines, such as zolpidem, zopiclone and zaleplon, demonstrate hypnotic efficacy similar to that of benzodiazepines along with excellent safety profiles. Non-benzodiazepines generally cause less disruption of normal sleep architecture than benzodiazepines. Psychomotor and memory impairment may be less problematic with non-benzodiazepines, especially when compared to longer-acting benzodiazepines. Rebound insomnia and withdrawal symptoms occur infrequently upon discontinuation of non-benzodiazepines and may be less common and milder than those seen upon discontinuation of some benzodiazepines. For the long-term treatment of insomnia, which is generally not recommended, zolpidem and zopiclone are particularly good options because they do not develop tolerance rapidly and have a low abuse potential. Limited data indicate that zaleplon has low tolerance and abuse potential, although further experience is needed to determine its long-term efficacy and safety profile. Since non-benzodiazepines produce minimal respiratory depression, they may be safer than benzodiazepines in patients with respiratory disorders. The choice of which hypnotic to use should be based on the patient's primary sleep complaint, health history, adverse effects and cost. PMID- 12531037 TI - The genetics of sleep apnea. AB - Obstructive sleep apnea hypopnea syndrome (OSAHS) is a complex chronic condition that is undoubtedly influenced by multiple factors. Accumulating data suggest that there are strong genetic underpinnings for this condition. It has been estimated that approximately 40% of the variance in the apnea hypopnea index (AHI) may be explained by familial factors. It is likely that genetic factors associated with craniofacial structure, body fat distribution and neural control of the upper airway muscles interact to produce the OSAHS phenotype. Although the role of specific genes that influence the development of OSAHS have not yet been identified, current research in rodents suggests that several genetic systems may be important. In this chapter, we shall first define the OSAHS phenotype, and then review the evidence that suggests an underlying genetic basis of OSAHS, the risk factors for OSAHS that may be inherited, and potential candidate genes. PMID- 12531038 TI - Treatment of sleep disturbance in Alzheimer's disease. AB - The prevalence of Alzheimer's disease (AD) is rapidly increasing as growing numbers of people around the world are living to old age. Sleep disturbances are a common, and often highly disruptive, behavioral symptom associated with AD. Nevertheless, the study of sleep in AD is relatively new. Little is known about the moderating factors that may alter a given patient's risk for developing sleep problems, or that may influence severity of presentation and persistence. Current treatments for improving sleep in AD fall into three broad categories: (i) pharmacological; (ii) cognitive-behavioral or psycho-educational strategies; and (iii) biological/circadian therapies. There are few studies demonstrating the efficacy of these treatments with community-dwelling AD patients, although studies with persons in institutional settings are promising. In this review, it is suggested that sleep problems in AD are multi-factorial, and influenced by a variety of demographic, physical, psychiatric and situational factors. These factors vary in how readily they can be modified and in how relevant they are to any individual case. Thus, when developing a treatment plan for sleep problems in a dementia patient, it is important to evaluate the underlying causes as well as the context in which the problems are occurring, and to target the intervention accordingly. PMID- 12531039 TI - Expanding horizons. PMID- 12531040 TI - Therapeutic options in obstructive sleep apnoea: have we made enough progress? PMID- 12531041 TI - Optimal continuous positive airway pressure for the treatment of obstructive sleep apnea/hypopnea. AB - The obstructive sleep apnea/hypopnea syndrome (OSA/H) is characterised by repetitive obstruction of the upper airway during sleep. The consequences of OSA/H include excessive daytime sleepiness, reduced perceived health quality and an increased risk of driving accidents. There is evidence that treatment with nasal continuous positive airway pressure (CPAP) is effective in eliminating these consequences. In addition, OSA/H may also be associated with hypertension and an increased risk of vascular disease, and although there is no clear evidence in this regard, CPAP may be effective alleviating these consequences as well. Nasal CPAP is considered the treatment of choice for clinically significant OSA/H. The optimal CPAP prescription is that which is most effective in eliminating the adverse consequences of OSA/H. Patients should be initially treated with standard CPAP. For those in whom standard CPAP is unsuccessful or intolerable bi-level or self-adjusting modes may be attempted. The nasal CPAP pressure level can be titrated to eliminate apnea, hypopnea, snoring, respiratory arousal and inspiratory flow limitation as a means of predicting successful long term therapy. The reliability of these physiologic parameters in predicting the optimal long-term CPAP pressure is eroded by the tendency of CPAP pressure-level requirement to decrease with chronic CPAP use, and by other factors that may increase the CPAP pressure requirement. The split-night polysomnogram and unattended auto-CPAP titration have been proposed as efficient means of determining the optimal CPAP pressure, but for patients with subtle OSA/H, underlying cardiopulmonary disease or other forms of sleep-disordered breathing a full-night polysomnogram may be required. The CPAP prescription must include a comfortable well-fitting interface. When CPAP is prescribed it is essential that the patient undergo clinical re-evaluation to ensure that treatment goals are being met. PMID- 12531042 TI - What's new in oral appliances for snoring and sleep apnea: an update. AB - The number and kind of oral appliances have proliferated in recent years as dentists have responded to the challenge to develop more sophisticated devices for the control of socially disruptive snoring and obstructive sleep apnea. Sleep disordered breathing is now recognised as being a highly prevalent problem which carries increased social, economic, cognition and health risks. The popular mandibular-advancement devices have a greater level of acceptance by patients than the standard nasal continuous positive airway pressure (CPAP) treatment in recent head-to-head studies. It is important that these patients be followed by sleep clinicians as these appliances are less successful in controlling sleep apnea when the level of severity is high. Research is needed to determine the patients most appropriate for an oromandibular treatment and when CPAP is the treatment of choice. Potential sequences of treatments are suggested. PMID- 12531043 TI - Medico-legal aspects of sleep disorders: sleepiness and civil liability. AB - Excessive sleepiness is associated with motor vehicle accidents and is responsible for enormous social and financial loss. The specific legal obligations for an individual with a sleep disorder, their employer and those health care practitioners associated with that individual are reviewed. Although there are related implications within the criminal law and in particular criminal negligence, the arguments developed in this paper will be largely confined to the context of the civil liability. The legal concepts of foreseeability and proximity are discussed in the context of sleep-related accidents. The reasoning of a recent Australian High Court judgement is discussed in view of the differences in legal and medical opinion on the extent of foreseeability of accidents as a result of sleepiness. Many countries have legislation designed to protect employees from injury at work and to protect the general public from injury. What is not clear is the extent to which an employer will be required to accept liability for an employee's sleepiness and the duty to monitor the health of their employees. Factors which influence this liability include: the extent to which the implications of the condition is known and understood generally; the extent to which the condition is suspected or identified in an individual employee; the extent of a proper screening and treatment program and the way in which risk management programs have been implemented. Although the issue of sleepiness and civil liability is examined from an Australian legal context, the principles have direct relevance to other legal systems. The authors highlight the degree of uncertainty provided by the common law and statutory provisions, and that decisions rest on the balance of public interests, which mean that many of the current dilemmas facing practitioners may only be solved in the courts. PMID- 12531044 TI - Secondary insomnia: diagnostic challenges and intervention opportunities. AB - The assessment and treatment of secondary insomnia (SI), the most common form of insomnia, are often complicated. Establishing an accompanying disorder as causal rather than comorbid is the key to assessment, but can be difficult even for experienced clinicians. Treatment often focuses on the primary disorder. In many cases, however, there is reason to treat the insomnia directly (insomnia is partially independent, does not respond to treatment of the primary condition, or has been misdiagnosed as SI). Although hypnotic medications are frequently used, behavioral interventions may be the best treatment approach, providing better long-term management of symptoms. Older adults warrant special consideration as age-related illnesses, changes in drug absorption and metabolism, and polypharmacy make them especially susceptible to SI. Recent research suggests successful treatment of insomnia may also relieve the primary disorder and merits follow-up. Future research is also needed on the efficacy of psychological treatment for SI for specific disorders as well as for older adults. PMID- 12531046 TI - Ceiling and floor effects in sleep research. PMID- 12531045 TI - Brain structures and mechanisms involved in the generation of REM sleep. AB - This article reviews the central nervous mechanisms involved in the broad network that generates and maintains REM sleep. Experimental investigations have identified the pontine tegmentum as the critical substrate for REM sleep mechanisms. Several pontine structures are involved in the generation of each particular polygraphic event that characterizes REM sleep: desynchronization in the electroencephalogram, theta rhythm in the hippocampus, muscle atonia, pontogeniculooccipital waves and rapid eye movements. The pontine tegmentum also holds the region where cholinergic stimulation can trigger all the behavioural and bioelectric signs of REM sleep. The exact location has been investigated and amply discussed over the last few years. Studies in the authors>> laboratory, mapping the pontine tegmentum with small volume carbachol (a cholinergic agonist) microinjections, have demonstrated that the executive neurons for REM sleep generation are neither located in the dorsal part of the pontine tegmentum, nor diffusely spread through the medial pontine reticular formation: they are concentrated in a discrete area in the ventral part of the oral pontine reticular nucleus (vRPO). In turn, the vRPO has connections with structures involved in the generation of the other states of the sleep-wake cycle as well as with structures responsible for the generation of each of the different events characterizing REM sleep. This allows us to propose the vRPO as the crucial region for REM sleep generation. Related research, with invivo and invitro experiments, into the actions of different neurotransmitters on vRPO neurones indicates that not only acetylcholine but other neurotransmitters have an active key role in vRPO REM sleep generation mechanisms. PMID- 12531047 TI - Sleep: big brain meets little brain. AB - 2001 Harcourt Publishers Ltd PMID- 12531048 TI - Gastrointestinal functioning during sleep: a new horizon in sleep medicine. AB - A body of clinical and research literature is accumulating suggesting that there are significant alterations in gastrointestinal functioning during sleep, as well as sleep complaints and disorders in patients suffering from gastrointestinal disease. This review addresses the clinical applications of some basic alterations in gastrointestinal functioning during sleep, with a particular focus on gastroesophageal reflux disease and functional bowel disorders. Recent studies have shown that gastroesophageal reflux during sleep results in a marked prolongation of esophageal acid clearance time, and consequent mucosal damage. Data are reviewed which suggest that the more serious complications of gastroesophageal reflux, e.g. esophagitis and the extra-esophageal complications of reflux such as the exacerbation of bronchial asthma, laryngopharyngitis, and pulmonary aspiration are the result of sleep-related gastroesophageal reflux. Recent studies have also shown that patients with functional bowel disorder (e.g. irritable bowel syndrome and dyspepsia) have a high incidence of sleep complaints as well as abnormalities of autonomic functioning. Recent studies have shown that the measurement of autonomic functioning during sleep can differentiate the patients with functional bowel disorders from normal controls. The continued study of gastrointestinal functioning during sleep clearly establishes a new horizon of investigation in both sleep medicine and gastroenterology. 2001 Harcourt Publishers Ltd PMID- 12531049 TI - Outcome measurement in sleep medicine practice and research. Part 1: assessment of symptoms, subjective and objective daytime sleepiness, health-related quality of life and functional status. AB - The documentation of treatment outcomes has gained increased importance as those responsible for paying for healthcare focus on the delivery of cost-effective care. The practitioner and researcher, challenged with documenting the clinical significance of their endeavors, must depend on conceptually appropriate, valid and reliable instruments. Outcomes of particular relevance for sleep disorders include self-reported general symptoms, subjective and objective daytime sleepiness, neurobehavior, health-related quality of life (HRQL) or functional status, incidence of accidents and cardiovascular morbidity, and health care utilization and cost. This paper will limit its focus to the outcomes of self reported general symptoms, subjective and objective daytime sleepiness, and HRQL or functional status. In addition to describing measures that could be applied to evaluate these outcomes, this paper will also discuss factors affecting the selection of outcome measures for use in research and practice, and consider evidence generated with the application of these instruments in research. Reductions in self-reported symptoms, such as snoring and daytime hypersomnolence following treatment, have been documented with the application of measures with strong psychometric properties such as the Survey Screen for Sleep Apnea and Epworth Sleepiness Scale. In addition to statistical reliability, calculation of the effect size and standardized response mean indicate that these changes are also clinically meaningful. Similar improvements have also been demonstrated using the Multiple Sleep Latency Test (MSLT), although the resulting increased latency has not typically been within normal values. Important improvements following treatment have been documented with the Maintenance of Wakefulness Test (MWT). Evidence suggests that the MWT may be more sensitive in situations, such as narcolepsy, where there are high levels of physiologic sleepiness, reserving the MSLT where the physiologic sleep tendency is more restricted, such as in obstructive sleep apnea (OSA). Patients>> perceptions of the benefit of treatment to their daily lives have been documented by such generic measures of HRQL, such as the SF-36 and Nottingham Health Profile. However, the impact of sleep disorders and the benefit of treatment on those aspects of daily behavior likely to be most affected have more clearly been demonstrated with the utilization of disease-specific HRQL measures such as the Functional Outcomes of Sleep Questionnaire and the Calgary Sleep Apnea Quality of Life Index. 2001 Harcourt Publishers Ltd PMID- 12531050 TI - Sleep disturbances after non-cardiac surgery. AB - After major non-cardiac surgery sleep pattern is usually disturbed with initial suppression of rapid eye movement sleep with a subsequent rebound during the first post-operative week. Deep sleep is also suppressed for several days after the operation and subjective sleep quality is impaired. The sleep disturbances seem to be related to the magnitude of trauma and thereby to the surgical stress response and/or post-operative opioid administration. Post-operative sleep disturbances may contribute to the development of early post-operative fatigue, episodic hypoxaemia, haemodynamic instability and altered mental status, all with a potential negative effect on post-operative outcome. Minimizing surgical trauma and avoiding or minimizing use of opioids for pain relief may prevent or reduce post-operative sleep disturbances. Post-operative sleep pattern represents an important research field, since it may have a significant negative on post operative outcome. 2001 Harcourt Publishers Ltd PMID- 12531051 TI - The duality of sleeping sickness: focusing on sleep. AB - Sleeping sickness, once under control, is a re-emergent endemic parasitic disease in intertropical Africa. Its originality resides in its duality. Two trypanosome groups (Trypanososma brucei gambiense vs.rhodesiense ) are transmitted to humans by tsetse flies from two geographical areas (Western and Central Africa humid forest vs. Eastern Africa arboreous savannah), provoking a slowly or a rapidly evolutive disease. The two stage (haemolymphatic vs. neurological invasion) pathogenic evolution leads to the duality of the immune response, depending on the host-parasite inter-relation differences in the blood and the brain. In the blood, the immune processes involved are both specific (anti-variant surface glycoprotein (VSG) antibodies) and non-specific (complement-mediated lysis, opsonification-facilitated phagocytosis and antibody dependent cell-mediated cytotoxicity). Although macrophages are activated in the blood and infiltrate the brain, nitric oxide decreases in the blood and increases in the brain, with a breakage in the blood-brain barrier, leading to brain lesions through the production of deleterious molecules. Prophylactic means are affected by the duality of pathogenic processes. This finally leads to a two stage disease (haemolymphatic vs. neurological) with two different therapeutic strategies. The sleep-wake cycle and other biological rhythms are also marked by the disappearance of circadian rhythmicity demasking basic ultradian activities and relationships, such as the interdependence of endocrine profiles and the sleep wake alternation. 2001 Harcourt Publishers Ltd PMID- 12531052 TI - How much sleep do we need? AB - There is increasing concern for sleeplessness-related risks in modern society. Some recent epidemiological data seem to support the view that many segments of the adult population have chronically inadequate sleep. On the other hand, some experts have claimed that our core, basic amount of sleep is around 6 h per night, and that the rest of our sleep can be easily curtailed, being unnecessary to fulfill any sleep need. However, experimental data on the effects of both acute and cumulative partial sleep deprivation (PSD) consistently point out that sleep restriction has substantial negative effects on sleepiness, motor and cognitive performance and mood, as well as on some metabolic, hormonal and immunological variables. As chronic PSD may have serious long-term adverse health effects, it should be avoided in the general population. In the short-term, the effects of sleep curtailment seem to accumulate linearly, while the effects of long-term PSD should be further investigated, as the few available studies are flawed by methodological weaknesses. On the other hand, there is evidence that extending sleep by 2-3 h beyond the norm produces only marginal benefits for an average individual. Finally, it is underlined that, as large individual differences do exist in the need for sleep, the search for the sleep need may be vain. A somnotypology, taking into account age, gender and the position in both the sleep-alert and the morningness-eveningness continuum, should help in the search for the actual individual sleep need. 2001 Harcourt Publishers Ltd PMID- 12531054 TI - Global aspects of tuberculosis in children. AB - Tuberculosis (TB) in children reflects the prevalence of the disease in adults as well as current transmission rates. Africa and South-east Asia have the largest number of tuberculosis cases and the situation there has been worsened by the HIV epidemic. Children born to HIV-infected parents, whether infected or not, are at high risk of developing tuberculosis because of the increased risk of exposure to the disease. Tuberculosis is more common among the disadvantaged and vulnerable groups in each society and the impact of overcrowding, under-nutrition and poverty is particularly severe on children. Recent studies have documented the increase in the occurrence of tuberculosis in children, both in developed and developing countries. The peak age of notification of tuberculosis decreases as the incidence of the disease increases in the region. Tuberculosis infection can progress rapidly to disease, particularly in infancy and early childhood. Most of the morbidity occurs in the first few years after infection. Recently infected children and those with large tuberculin reaction (>18 mm) are at increased risk for progression and should be followed closely. Mortality from tuberculosis is also highest in early childhood, mainly due to disseminated forms like meningeal and miliary tuberculosis. Tuberculosis can be controlled either by preventing the infection from occurring or by treating early infection after it has occurred. An efficient tuberculosis control program with early detection of infectious adults and their cure is the best long-term approach to the reduction of TB disease in children. The DOTS strategy advocated by the WHO has the potential to have a significant impact on the epidemiology of tuberculosis by achieving high cure rates and thereby decreasing community transmission. BCG vaccination, through effective against disseminated forms of the disease in childhood, has very little impact on adult forms of the disease. Chemoprophylaxis or preventive therapy is effective, but difficult to implement on a mass scale and is only recommended for special high-risk groups in developing countries. PMID- 12531055 TI - Childhood tuberculosis: public health and contact tracing. AB - Contact tracing is extremely useful to detect patients with latent tuberculosis (TB) infection and should be followed by preventive therapy. When properly applied it can have an impact on morbidity and mortality, especially among children. In most industrialised countries the decline in the risk of infection due to better overall health and effective TB control measures is likely to result in the disappearance of tuberculosis as a public health problem; therefore the cost-benefit of contact tracing is likely to be less than in the developing world. However, given an infection prevalence of up to 40% in adolescence and the more immediate need to target scarce resources at treating active infectious cases, developing countries cannot extend the practice of contact tracing and preventive therapy beyond the very young and groups at high risk. In developing countries more benefits are likely to arise from widespread and effective application of the DOTS strategy for the prevention and the control of tuberculosis in both adults and children. Diagnosis and treatment of known infectious cases must necessarily precede a recommendation for contact tracing, however great the perceived impact may be. In the interim, political commitment to ensure global equality in accessing health care, research to improve screening procedures, lower costs of treatment through shorter regimens and the application of comprehensive measures for tuberculosis prevention and control are desperately needed. PMID- 12531056 TI - Childhood tuberculosis: treatment strategies and recent advances. AB - Over the past 20 years, several major studies have shown that 6-month therapy, initially using isoniazid, rifampin and pyrazinamide, is highly effective and extremely safe for the treatment of most forms of childhood tuberculosis. The various drug schedules and frequency of administration will be reviewed. Directly observed therapy is an essential component of a paediatric tuberculosis treatment plan, though using it does not solve all problems with adherence to treatment. As the rates of drug-resistant tuberculosis increase around the world, special aspects of paediatric tuberculosis will have to be considered when designing treatment regimens for children. Finally, the next frontier of antituberculosis therapy may be the manipulation of the host immune system. PMID- 12531057 TI - Non-pulmonary tuberculosis. AB - Tuberculosis (TB) is a serious disease of global importance, with a rising incidence in the developed world in recent years. Tuberculous lymphadenitis, tuberculous meningitis, osteoarticular tuberculosis and miliary tuberculosis are some of the more well-recognised manifestations of non-pulmonary TB in childhood. The diagnosis of non-pulmonary TB poses a particular challenge for clinicians because of the protean ways in which the disease presents. The omission of tuberculosis from the differential diagnosis of patients with obscure illnesses and the relatively insensitive bacteriological methods for detecting Mycobacterium tuberculosis add to the complexity of the problem. A high index of suspicion is required in order to avoid delays in diagnosis which may influence treatment outcome. The advent of DNA amplification techniques such as the polymerase chain reaction may herald a promising new era in the prompt and accurate management of extrapulmonary tuberculosis. PMID- 12531058 TI - Diagnostic techniques in paediatric tuberculosis. AB - Accurate diagnosis of tuberculosis (TB) in children is crucial to provide effective treatment and to identify the undiagnosed adult who probably infected the child. The diagnosis of tuberculosis in children is often based only on epidemiologic, clinical, and radiographic findings. Fibre-optic bronchoscopy may also be helpful but the recovery of tuberculous bacilli, which establishes the diagnosis, is difficult in children. Rapid diagnostic tests such as direct amplification and serological tests have been developed to improve early diagnosis. Their sensitivity is higher than that of conventional mycobacterial culture in most paediatric studies. PMID- 12531059 TI - Clinical applications of infant lung function testing: does it contribute to clinical decision making? AB - Infant lung function testing is important in clinical research and recent standardisation efforts have enabled measurements to be made in infants in different laboratories throughout the world. Thus, the theoretical conditions are now fulfilled for use of these techniques in clinical practice. This review discusses the usefulness of various infant lung function techniques in a clinical setting and their potential present or future clinical application. It will focus on the role of infant lung function in the clinical management of infants with respiratory disease. PMID- 12531060 TI - Thoracic interventional radiology in children. AB - Significant technological advancements have been achieved in radiology with the introduction of a branch speciality known as interventional radiology. Radiology has moved into the therapeutic arena, performing minimally invasive diagnostic and therapeutic procedures. The interventional radiologist can treat certain vascular and non-vascular conditions with "keyhole" or perhaps more correctly "pinhole" techniques using catheter-based technology directed under image guidance with X-ray (including computed tomography), ultrasound and magnetic resonance imaging. Interventional radiology can provide new treatment options for children not possible a few years ago. PMID- 12531061 TI - Molecular mechanisms of corticosteroid actions. AB - Glucocorticoids are the most effective therapeutic agents used for the treatment of chronic inflammatory diseases of the lung. They act by interacting with, and thereby activating, a specific cytoplasmic receptor (GR) which then migrates to the cell nucleus and inhibits inflammatory gene transcription. Inflammatory gene transcription is enhanced by activation of several intracellular signalling pathways which activate transcription factor such as NF-kappaB and AP-1. These transcription factors bind to DNA and induce local unwinding of the DNA structure allowing increased gene transcription. Glucocorticoids interfere with the ability of NF-kappaB and AP-1 to induce transcription by increasing the compaction of unwound chromosomal DNA in a process that involves deacetylation of histone proteins. A small number of asthmatic patients are resistant to the beneficial effects of glucocorticoids. This defect does not appear to be due to a reduced expression of GR but may be associated with a failure of GR to translocate into the nucleus and/or a reduced ability to inhibit AP-1 actions. PMID- 12531062 TI - Surfactant proteins: role in lung physiology and disease in early life. AB - Pulmonary surfactant is an amalgam of proteins and phospholipids which serves to maintain a low surface tension within the alveolar regions of the lungs during changes in lung volume. Recently, two of the surfactant proteins--A and D--have been characterised within the collectin family and found to play important roles in the non-specific host defence of the lung. The field of surfactant biology has attracted the attention of physiologists, biochemists, molecular biologists and clinical scientists in an effort to describe the nature and role of pulmonary surfactant in health and disease. This paper will review the history and content of discoveries in the field of surfactant biology together with pulmonary diseases related to surfactant deficiency or dysfunction. PMID- 12531063 TI - Therapeutic strategies to correct malfunction of CFTR. AB - Cystic fibrosis (CF) is a systemic autosomal recessive inherited disorder that results from mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Although the gene was cloned 11 years ago, there still is no definitive treatment to correct the functional deficit. Current treatment strategies focus on pancreatic enzyme replacement and control of pulmonary inflammation and infection. This review examines novel strategies still in preclinical development or phase 1 clinical trials. Gene therapy is an evolving area of study that offers the potential for a cure for cystic fibrosis. CF lung disease is a significant barrier to effective gene delivery and transfer, but new vectors show promise in overcoming these limitations. There are also new pharmacological therapies aimed at correcting defects in CFTR processing and function. These are tailored to the specific class of mutation but may offer therapeutic benefit to many patients. They include phenylbutyrate, flavonoids, aminoglycosides and xanthines. PMID- 12531064 TI - Education, self-management and home peak flow monitoring in childhood asthma. AB - Current asthma guidelines focus on self-management by the patient, in which monitoring of peak flow plays an important role. To be able to participate in self-management, the patient must be educated rigorously on pathophysiological mechanisms of the disease, principles of treatment, correct inhalation technique, treatment goals and the action to take when symptoms or peak flow worsen. This is a time-consuming but important and worthwhile task. The pivotal role of home peak flow monitoring in asthma self-management is based on the assumptions that peak flow variation is a useful measure of disease activity and that peak flow diaries are kept reliably by patients. There is now evidence that neither of these assumptions are true. Self-management plans based on education alone are just as effective as those incorporating peak flow monitoring. Education, therefore, is the most important component of asthma self-management, and home peak flow monitoring is not needed in the majority of asthmatic children. PMID- 12531065 TI - Guest editorial. PMID- 12531066 TI - Pulmonary neuroendocrine cells (PNEC) and neuroepithelial bodies (NEB): chemoreceptors and regulators of lung development. AB - The airway and alveolar epithelia contain pulmonary neuroendocrine cells whose structure indicates an endocrine function. They are also in contact with sensory nerve fibres. These cells often aggregate into distinct corpuscles neuroepithelial bodies-and carry membrane receptors sensitive to a number of stimuli, including hypoxia and nicotine. They synthesise, store and release a number of bioactive substances such as serotonin, calcitonin gene-related peptide and the mitogen bombesin. When these are released they contribute to redistribution of pulmonary blood flow, regulation of bronchomotor tone, modulation of the immune response, stimulation of sensory nerve fibres and regulation of lung growth and development. Pulmonary neuroendocrine cells and neuroepithelial bodies seem to be most important in the fetal and neonatal lung as regulators of airway development and hypoxia-sensitive chemoreceptors. There is a link between these cells and specific types of lung cancer and their involvement in lung and paediatric pathology may be profound. PMID- 12531067 TI - Guest editorial. PMID- 12531068 TI - Genetic/environmental determinants of adult chronic obstructive pulmonary disease and possible links with childhood wheezing. AB - Epidemiological evidence and similarities in underlying inflammatory mechanisms suggest that childhood respiratory conditions and adult onset chronic obstructive pulmonary disease (COPD) may have a common aetiology. The prevalence of COPD in the UK has been estimated to be 10% in males and 11% in females, and like asthma, is thought to have a polygenic contribution with a complex interplay between genetic and environmental factors. The only proven genetic factor so far identified in its pathogenesis is alpha1-antitrypsin deficiency, although this accounts for less than 1% of individuals with disease. Environmental exposures are significant but the observation that only 15% of smokers develop COPD and only some young children exposed in utero develop recurrent wheezing points to as yet undefined contributory genetic factors. The similarity in response to medication also suggests some features in common and a central question must be whether those adults who develop COPD had transient viral-associated wheeze (VAW) or non-specific bronchial hyperresponsiveness (BHR) in childhood? Similarities between some of the childhood wheezing syndromes and adult onset COPD invites a re-examination of possible links between the two conditions. PMID- 12531069 TI - Recent advances in intubation and mechanical ventilation. AB - The ideal care of the critically ill infant or child should demonstrate a seamless service starting with recognition of potential critical illness and initiation of early resuscitative interventions, escalating to advanced life support and skilled transfer to an appropriate location where intensive care can be continued. This article discusses advances in the package of respiratory care commencing with the decision to provide respiratory support, which includes the decision whether or not to intubate, leading to the choice of mechanical ventilatory support mode in the critically ill child. Indications for intubation include protection of the airway, airway toilet and the need for positive pressure support. Respiratory support may not always require additional airway protection or airway toilet. Non-invasive respiratory support includes continuous positive airway pressure, biphasic positive airway pressure, or negative extrathoracic pressure. Recent advances in understanding and management of acute lung injury, such as appropriate use of positive end-expiratory airway pressure, alveolar recruitment manoeuvres, high frequency ventilation and use of inhaled nitric oxide, will be discussed in the light of recent randomised controlled trials providing evidence of benefit. PMID- 12531070 TI - Case 1: presentation. Mycobacterium tuberculosis. PMID- 12531071 TI - Case 2: presentation. Poor response to asthma treatment. PMID- 12531072 TI - Case 3: presentation. Dyspnoea without wheeze unlikely to be caused by asthma. PMID- 12531073 TI - Case 4: presentation. Asthma or viral-associated wheeze. PMID- 12531074 TI - Case 5: presentation. Pulmonary tuberculosis. PMID- 12531081 TI - Basic epidemiology and immunopathology of RSV in children. AB - Respiratory syncytial virus (RSV) is the most important cause of lower respiratory tract infection in infants and young children. Around 20 000 RSV infected infants require hospitalization in the UK during each yearly epidemic, which is about 3% of the birth cohort. Most children are infected by 2 years of age. Risk factors for severe disease include young infants, prematurity, chronic lung and cardiac conditions or immunodeficiency. Humoral immunity is incomplete and short-lived, yet reinfections cause less severe disease. RSV infects infants despite the presence of specific neutralizing antibodies. RSV infection can be linked to the development of individual wheezing episodes. A competent cellular immune system is necessary to reduce disease severity. RSV infection provokes an RSV-specific T-lymphocyte response with the release of cytokines. There is a delicate balance between the protective and disease-enhancing effects of the host's immune response to RSV infection. PMID- 12531082 TI - RSV: Management of the acute episode. AB - Bronchiolitis is the most common lower respiratory tract infection in infants and is responsible for the majority of paediatric hospital admissions in the winter. Despite significant advances in pharmacotherapy, the management of infants with bronchiolitis has changed little over the years from supplemental oxygen and good fluid management. This paper reviews current treatment options for bronchiolitis, including the use of bronchodilators, adrenaline, steroids and ribavirin. More recent advances, including immunotherapy and intensive care, are discussed. PMID- 12531083 TI - Long-term consequences of respiratory syncytial virus (RSV) bronchiolitis. AB - Despite differences in study design, follow-up studies consistently show that approximately half of the infants with respiratory syncytial virus (RSV) lower respiratory tract infection (LRTI) go on to have recurrent wheezing episodes during childhood. Respiratory symptoms are associated with abnormal lung function, including bronchial hyper-responsiveness. Wheezing symptoms following RSV LRTI gradually decrease, and it appears that during school age airway morbidity is no longer related to RSV LRTI during infancy. Mechanisms underlying the association between RSV LRTI and long-term airway morbidity are poorly understood. On the one hand, abnormal airway function that is congenitally present or acquired before RSV LRTI occurs could be the cause of both RSV LRTI and subsequent recurrent wheezing. On the other hand, it is possible that RSV LRTI causes changes in the lower airways or the immune system that result in long term airway morbidity. Animal models suggest that RSV infection can promote the development of allergic sensitization, but most studies in humans do not indicate a role for atopy in the development of recurrent wheezing following RSV LRTI. PMID- 12531084 TI - Prevention of bronchiolitis. AB - This review evaluates the current situation and long-term prospects for containment of human respiratory syncytial virus (HRSV) infection and bronchiolitis in infancy. The biology and immunopathology of HRSV infection are complex. Initial attempts to control HRSV infection using a conventional formalin inactivated vaccine had the unexpected outcome that the disease was potentiated in some vaccinees experiencing natural HRSV infection at a later date. Much effort has been devoted to defining the nature of protective immunity, and several candidate sub-unit and live attenuated vaccines have been developed by empirical and semi-empirical routes, and most recently by reverse genetics. None has yet received approval for clinical use, and attention has switched from active to passive immunization. Both concentrated human immune globulin (RespiGam) and a humanized monoclonal antibody (Palivizumab) have been approved for clinical use. On grounds of cost-effectiveness these treatments are recommended only for treatment of high-risk infants. An effective antiviral is not yet available. PMID- 12531085 TI - Cellular and humoral defence mechanisms against bacteria. AB - Host defence to bacterial infection is a complex process of highly efficient effector systems that co-operate to eliminate microbial invaders. The systems not only co-operate, they are also interdependent, as deficiency of one system may cause dysfunction in another. As is so often the case in medicine, the discovery of the various effector systems and their components originated from studies of patients with unusually severe, frequently recurring infections. Modern molecular biology has greatly aided in elucidation of the basic disorders and identification of the corresponding gene defect. This knowledge will hopefully provide us with better tools for causal therapy in the future; at present the only available "cure" is bone marrow transplantation, a treatment modality with many problems of its own. PMID- 12531086 TI - A practical approach to the interpretation of lung function testing in children. AB - Lung function testing is an increasingly important tool in the diagnosis and monitoring of lung disease in children. However, not all paediatricians will have had training in lung function testing. Here, a structured approach to the interpretation of lung function results is developed. This should help the clinician understand the results from their lung function laboratory. PMID- 12531087 TI - Magnetic resonance imaging of the paediatric chest. AB - Magnetic resonance (MR) has an important role to play in the imaging of the paediatric chest, not least because of its zero ionizing radiation dose. Computerized tomography (CT) has been the preferred technique for cross-sectional imaging to date because of the ease of access, speed and superior spatial resolution. This article discusses the clinical situations where magnetic resonance may be used as the primary cross sectional imaging modality. The clinical indications and the complimentary roles of magnetic resonance and computerized tomography are discussed. The patient preparation, technical aspects, advantages and disadvantages of MR are documented. PMID- 12531088 TI - Inflammatory mediators in blood and urine. AB - Inflammation plays a major role in the pathophysiology of asthma. Therefore, monitoring of the disease and its management preferably should include assessment of mediators of airways inflammation. Much interest has been focused on using eosinophil granule proteins in blood and urine as markers of asthma inflammation. The eosinophil granule proteins are important mediators in allergic inflammation. They often function well as inflammatory markers when used in controlled clinical studies, and are therefore useful research tools. With urinary eosinophil-derived protein X (EPX) venous blood sampling is avoided. Disadvantages which limit usefulness in clinical practice are overlap between values in patients and controls, a rather weak correlation to traditional lung function variables, elevation of levels by concurrent allergic disease, and the delay between sampling and test results. Urinary excretion of leukotriene E4 (LTE(4)) reflects the production of cysteinyl leukotrienes, which are major mediators in asthma. The excretion of LTE4 possibly reflects lung function better than serum ECP. In the future, monitoring of cytokines may be used in clinical asthma to monitor control and aid in the prognosis of the disease in the young child. Techniques for simple and rapid monitoring of key type 1 and type 2 cytokines are needed. PMID- 12531089 TI - Pulmonary haemorrhagic syndromes in children. AB - Pulmonary haemorrhage in children is rare and may be either focal or diffuse, idiopathic or associated with an underlying disease. Focal bleeding is usually associated with congenital anomalies or localized acquired disease. Many of the disorders causing diffuse alveolar haemorrhage appear to be immunologically mediated. Treatment modalities include management of the underlying disease and the use of corticosteroids, chloroquine and/or cytotoxic drugs and induce remission in more than half of the patients with diffuse haemorrhage. PMID- 12531090 TI - Atelectasis: mechanisms, diagnosis and management. AB - The term atelectasis describes a state of collapsed and non-aerated region of the lung parenchyma, which is otherwise normal. This pathological condition is usually associated with several pulmonary and chest disorders and represents a manifestation of the underlying disease, not a disease per se. Atelectasis may occur in three ways: (i) airway obstruction; (ii) compression of parenchyma by extrathoracic, intrathoracic, chest wall processes; and (iii) increased surface tension in alveoli and bronchioli. Chest radiographs using both the anterior posterior and lateral projections are mandatory to document the presence of atelectasis. Differentiation from lobar consolidation may be a clinical dilemma. The treatment of atelectasis varies depending on duration and severity of the causal disease from chest physiotherapy to postural drainage, bronchodilator and anti-inflammatory therapy. Persistent mucous plugs should be removed by bronchoscopy. PMID- 12531091 TI - Malignant disease and the lung. AB - Primary lung tumours in childhood are rare. However, cancer in a child may have an impact on the lung in a number of ways. Chemotherapy and radiotherapy may be directly toxic to the lung. Young children are particularly sensitive to the effects of radiotherapy, which can cause impairment of growth of muscle, skin and bone, in addition to its direct toxic effect on the underlying lung. The lung is vulnerable to infection - particularly protozoal, viral and fungal organisms, as well as bacterial. Children undergoing bone marrow transplantation are at greater risk of lung damage, as they are profoundly immunosuppressed and have received intensive cytotoxic chemotherapy or radiotherapy. The underlying cause of lung damage may be difficult to determine because of the complexity of treatment and the additional risk of infectious complications. In a small number of children, pulmonary complications may be fatal. However, for the many survivors, although abnormalities of lung function are frequently detected, these are rarely clinically significant and, with notable exceptions, do not appear to deteriorate with time. However, data remain scanty; there is a real need for ongoing prospective studies of lung function in survivors of childhood cancer. PMID- 12531092 TI - Evidence-based medicine. AB - Evidence-based medicine aims to identify, critically appraise and apply the best available evidence in making decisions about the care of patients. These aims are similar to those which conscientious clinicians have always tried to achieve. An evidence-based approach applies a systematic and rigorous methodology to this process to ensure that the evidence which is applied is relevant and of high quality. The use of this methodology limits bias (systematic errors) and reduces random errors (simple mistakes), thus providing reliable results on which to draw conclusions and make decisions. Meta-analysis, when used appropriately, can provide more precise estimates of the effects of healthcare than those derived from the individual studies included in a review. Systematic reviews also provide summaries of the results of evidence-based healthcare. These are available to all interested individuals, including clinicians, healthcare administrators and patients. PMID- 12531095 TI - The American Thoracic Society, Toronto, May 2000. PMID- 12531096 TI - American Thoracic Society Meeting, May 2000: The Genetics of Asthma. PMID- 12531100 TI - Cystic fibrosis: basic science. AB - Advances in the elucidation of cystic fibrosis transmembrane regulator (CFTR) function have resulted in a greater understanding of the relationship between the CF gene defect and clinical disease. The clinical phenotype is influenced by the class of mutation and possibly by other modifier genes. CFTR regulates the volume and composition of airways surface liquid, primarily by controlling chloride ion transport. However, CFTR also regulates other membrane channels and transports other molecules which may be important in mucocilary clearance and innate defence mechanisms. CFTR may also modulate the inflammatory response in respiratory epithelial cells and other inflammatory cells through a range of mechanisms. It is apparent that CFTR dysfunction results in a range of effects which may contribute to the clinical phenotype. These may contribute to the development of clinical disease at different stages of the natural history of cystic fibrosis. A greater understanding of the basic defect and its implications is likely to result in novel therapeutic approaches. PMID- 12531101 TI - Inflammation in cystic fibrosis and its management. AB - Pulmonary complications are by far the most serious complications in cystic fibrosis (CF). In response to endobronchial bacterial infections, the long-term consequences of chronically activated polymorphonuclear leukocytes (neutrophils) are thought to cause loss of lung function and premature death. Effective anti inflammatory therapy implies that we understand the complex networks of the immune response with its myriads of mediators on effector cells. We have many steps to go in this context, but science has already shed some light on inflammatory mechanisms in CF airways, and anti-inflammatory treatment strategies have been applied clinically with some success. The aim of this chapter is to review our present knowledge, and we will focus on early inflammation, bacterial persistence mechanisms, neutrophil activation, cytokines and nitric oxide together with anti-inflammatory treatment strategies in CF patients. PMID- 12531102 TI - Newer therapies for cystic fibrosis. AB - Cystic fibrosis (CF) is a chronic, progressive, genetic disease caused by flawed ion transport across epithelial membranes due to a genetic mutation. Most therapeutic efforts are centred on the main clinical manifestations of the disease: progressive destructive airway disease and pancreatic insufficiency. Most individuals with CF succumb to lung disease. The present-day therapeutic armamentarium includes agents that have been used for many decades, some of which have experienced transformations in their formulation or mode of administration thanks to the introduction of new manufacturing technologies. The development of new therapies involves new conceptual approaches, based on recent understanding of the disease. These therapies await proof of concept or clinical experimentation before being accepted as useful means to arrest the progression of the disease. In this article we will review therapeutic agents introduced into the clinical arsenal in the last 20 years, as well as experimental therapies under active investigation. PMID- 12531103 TI - Lung transplantation and end of life issues in cystic fibrosis. AB - Lung transplantation has been available as a therapeutic option for patients with end-stage cystic fibrosis lung disease for over 15 years, but the outcome following transplantation remains poor, and the supply of organs limited. For this reason some children opt to continue with medical treatment followed by terminal care rather than undergo transplantation. This article summarizes the current status of lung transplantation, including current referral guidelines and contraindications and also addresses current practice in terminal care. PMID- 12531104 TI - Cystic fibrosis: psychological issues. AB - To most parents the diagnosis of cystic fibrosis (CF) in their child represents a severe blow because they are confronted with an unwanted and unexpected disease that completely changes their whole life. Common reactions such as shock, denial, sadness and anger have to be mastered before a gradual adaptation to reality will be possible. The provision of support by a multidisciplinary team, including a psychologist, that offers its services from diagnosis and puts an emphasis on preventive care may help to achieve, maintain and improve physical and mental health, and social functioning in both patients and parents. CF not only affects the individual but the whole family, and the presence of biopsychosocial stressors may add to the burden caused by the disease. CF does not necessarily cause long-term serious family dysfunction, but it changes family structures and often taxes the family system beyond its strength. Even if there is only partial adherence to the demanding and complex treatment regimen, health professionals need to acknowledge the tremendous underlying effort on the part of the families. As a consequence of their continuous endeavours, many individuals with CF do lead remarkably normal lives with the prospect of gene therapy and lung transplantation, maintaining hope in case of severe deterioration. PMID- 12531105 TI - Host defence mechanisms of the respiratory system. AB - From the moment of the first breath at the time of birth, the respiratory system must be protected from numerous insults from the environment. The way in which the airways are prepared for these insults is by an extraordinarily effective defence system. This defence system includes mechanical as well as biochemical processes that work in an integrated way to safeguard the lungs. The airways have an efficient, highly vigilant, non-inflammatory anti-microbial defence system, capable of dealing with a wide spectrum of microbes, and provide lifelong protection of these vital mucosal surfaces. The different aspects of this defence system of the airways are discussed. The effectiveness of the system must be improved, especially in children prone to frequent respiratory infections like cystic fibrosis. Future research must be focused on restoring the defect in the defence mechanism, but also in a more appropriate shield against pathogenic micro organisms. PMID- 12531106 TI - Lung volume measurements in childhood. AB - Although less commonly used in clinical practice than spirometry, lung volume measurements are helpful in assessing progress and response to therapy in situations where lung volume is likely to be increased, as in asthma or cystic fibrosis, or reduced secondary to pulmonary hypoplasia, pulmonary fibrosis, or musculoskeletal abnormalities. Their contribution to research has been considerable. The techniques most commonly used are those of inert gas dilution on rebreathing, applicable to children of all ages but tending to under-record in the presence of gas trapping. The gold standard is body plethysmography. This is relatively expensive and requires more technical skill, but will measure all the gas within the chest. Computer programmes have been developed to obtain a reasonable estimate of lung volume from chest X-rays alone. More accurate results are obtained by echo-planar imaging but this is only available in highly specialized units. Information on fetal lung volumes can be obtained using digitized ultrasound images and position sensors attached to the ultrasound probe. PMID- 12531107 TI - HRCT of paediatric lung disease. AB - Thin-section or high-resolution computed tomography (HRCT) is used in children for the investigation of disorders such as airway disease and diffuse interstitial lung disease. Advances in CT technology have resulted in faster scan acquisition times, enabling images of sufficient diagnostic quality to be obtained in children during gentle respiration. Whilst radiation dose is an important consideration, the dose of a thin- section CT is approximately one seventh that of a conventional CT examination. As in adults, thin-section CT is more sensitive than chest radiography and may demonstrate abnormalities despite a normal chest radiograph. HRCT is also more specific than chest radiography for categorizing airway, airspace or interstitial processes in children. The current review will concentrate on the value of CT in airway diseases and diffuse interstitial lung disease. PMID- 12531108 TI - Bronchial hyper-responsiveness in children. AB - Bronchial hyper-responsiveness is a cardinal feature of asthma in children, but is also present in children with some other types of chronic lung disease. Techniques are now available that enable bronchial reactivity to be measured in children of almost all ages. The responses of normal and asthmatic children to standardized challenges have been measured and the optimal cut-off values separating the normal and asthmatic response have been established statistically. All types of challenge can distinguish between the child who is normal and one with lung disease within the limits of the sensitivity and specificity of the test. However, only indirect challenges, such as those by exercise or the inhalation of adenosine 5'-monophosphate, distinguish between asthma and other types of lung disease. PMID- 12531109 TI - Exotic pulmonary infections. AB - Pulmonary infections are the commonest type of illnesses seen in everyday practice. Although most infections are caused by a few familiar pathogens, an increase in global travel has led to the occurrence of rare and exotic diseases that often do not manifest until after the traveller returns home. Previously innocuous organisms may also be the cause of exotic infections in patients undergoing transplantation, or very aggressive immunosupression. Such treatment regimes may reactivate quiescent organisms acquired years previously, for example melioidosis and strongyloides stercoralis. In most developed countries there is also an increasing incidence of zoonotic infections, with a number of these presenting with mainly pulmonary symptoms. A carefully taken history of recent travel, exposure to wild or domestic animals or the presence of immunosuppression, is extremely helpful in identifying problematic infections that fall into the realm of the exotic. This review discusses some of the respiratory infections in each of these groups with an emphasis on imported infections. PMID- 12531110 TI - Non-invasive home ventilation. AB - Non-invasive ventilation (NIV) provides significant advantages to the paediatrician who cares for children with chronic ventilatory insufficiency and who would like to manage these children in the home environment. There is a growing experience in the use of NIV in the management of children with upper airway obstruction, ventilatory failure due to weakness or distortion of the thoracic cage and in children with abnormalities to their central respiratory drive especially during sleep. Success hinges on choosing the correct patient and the most appropriate device with a team approach to both the early assessment and follow-up. It may not be successful on the first attempt and often a phased introduction and some perseverance is required on the part of the paediatric team and the family. Home ventilation is possible, practical and advantageous compared with institutional living and non-invasive techniques can assist the paediatrician in achieving this goal. PMID- 12531111 TI - Chronic interstitial lung disease in children. AB - Chronic interstitial lung disease (ILD) is rare in children, but is clinically relevant because of its high morbidity and mortality rates. Several aetiological agents may have a role in this disease and various aetiopathogenic forms have been described, although they share a characteristic evolution towards pulmonary fibrosis. The aetiology sometimes remains unknown, in which case we speak of idiopathic interstitial pneumonia (IIP). Diagnosis relies on non-invasive methods (clinical history, respiratory function tests, chest X-ray and high-resolution CT scan) and invasive techniques (bronchoalveolar lavage, transbronchial biopsy, video-assisted thoracoscopic biopsy and open lung biopsy); the latter are used when the former fail to identify an aetiological agent. The gold standard for the diagnosis is lung biopsy, however, which also enables the classification of the idiopathic forms, in particular (usual interstitial pneumonia, desquamative interstitial pneumonia, non-specific interstitial pneumonia, and so on). In ILD, the most common therapeutic approaches involve the use of oral steroids and hydroxychloroquine sulphate for its anti-inflammatory and antifibrotic effect. The prognosis is often poor. PMID- 12531112 TI - Good care for people with cystic fibrosis. AB - The improvement in the health and survival of people who have cystic fibrosis (CF) has been due to better treatment developed at major CF centres. The regimens for prevention, early treatment and later stabilization of chronic respiratory infection and for the maintenance of normal nutrition and growth are now largely established. Treatment is life-long, complex and expensive. It should be started early after a diagnosis made following neonatal screening, and before chronic respiratory infection and malnutrition are established. Regular monitoring and input from the expert staff of a CF centre is essential, either on a 'full' or 'shared care' basis; adults with CF should attend a major Adult CF unit. The details of the staff and facilities necessary to achieve good care for CF are discussed, including the details of clinic procedures and annual assessments. PMID- 12531113 TI - Physiotherapy for children with cystic fibrosis. AB - Physiotherapy is an important part of the daily routine of patients with cystic fibrosis. It is one of a number of treatments used to help improve the quality of life. Airway clearance techniques aim to relieve respiratory symptoms by physical means, helping removal of tracheobronchial secretions, and thereby preventing or slowing deterioration in respiratory function. Exercise is used to monitor patient progress, maintain thoracic mobility, and improve well being and morale. Cystic fibrosis patients require ongoing assessment throughout life; physiotherapists can help to minimize the effect of the treatment burden by being as flexible as possible with regard to where, when and how they assess and plan treatments with their patients. PMID- 12531118 TI - Systemic and pulmonary haemodynamics during obstructive sleep apnoea. PMID- 12531119 TI - Systemic hypertension and obstructive sleep apnoea. AB - This article is a review of the current evidence that links systemic hypertension with obstructive sleep apnoea. Whilst a causal association has been suspected for some time, the day to day variability of both blood pressure and sleep apnoea severity, and clustering of confounding cardiovascular risk factors in sleep apnoea patients has made this association difficult to prove. There is unassailable evidence that obstructive apnoeas raise blood pressure acutely in both animal models and humans, through a combination of autonomic and state dependent arousal with some mechanical influences, and these rises can be controlled by nasal continuous positive airway pressure. Thus, although repetitive apnoeas alter the blood pressure variability and raise sleeping blood pressure in patients with OSA and sophisticated animal models have demonstrated increases in daytime blood pressure after the onset of OSA in the short term, such effects on diurnal BP have yet to be proven in humans. Recent rigorously designed large epidemiological studies have proven an independent association between OSA and systemic hypertension in both general and sleep clinic populations, with closely matched case control series also reporting raised blood pressure in OSA patients. A direct temporal causal association between the onset of obstructive sleep apnoea and raised blood pressure is expected to be confirmed by longitudinal data from the continuing epidemiological population studies. Finally, several studies on the beneficial effects of nasal continuous positive airway pressure in reducing blood pressure in OSA patients have preliminary results in abstract form, with one published in full. PMID- 12531120 TI - Pulmonary haemodynamics in obstructive sleep apnoea. AB - In patients with obstructive sleep apnoea syndrome (OSAS), pulmonary haemodynamics can show both transient perturbations during sleep and permanent alterations. During sleep, repeated fluctuations in pulmonary artery pressure and pulmonary wedge pressure, coincident with apnoeas, can be observed. Calculation of transmural pressure values is preferable to intravascular pressures in OSAS, due to the marked swings in intrathoracic pressure associated with obstructive apnoeas. Pulmonary artery pressure may progressively increase during sleep, particularly in close sequences of highly desaturating apnoeas. Apnoea-induced hypoxia appears as the most important determinant of this pulmonary artery pressure behaviour. Stroke volume and cardiac output during obstructive apnoeas show changes mainly related to intrathoracic pressure variations. Permanent precapillary pulmonary hypertension at rest is observed in <50% OSAS patients, and is poorly reversible after OSAS treatment. It correlates best with diurnal respiratory function parameters. However, the finding of pulmonary hypertension in some patients with near normal diurnal lung function led to suggest that sleep respiratory disorders may contribute to permanent pulmonary haemodynamic impairment in predisposed subjects. Knowledge on right ventricle hypertrophy in OSAS is inconsistent. As to right ventricle failure, it is clinically evident in subjects with associated lung disease or morbid obesity, while it may be detected instrumentally in subjects without such alterations, presumably as effect of apnoeas themselves. Besides, it appears more fully reversible after long-term OSAS treatment than pulmonary hypertension. PMID- 12531121 TI - Upper airway surgery for obstructive sleep apnea. AB - Upper airway surgical treatments for obstructive sleep apnea syndrome (OSAS) attempt to modify dysfunctional pharyngeal anatomy or by-pass the pharynx. Modifications of the pharynx diminish the bulk of soft tissue structures which abut the air column, place them under tension, others alter their spatial inter relationships. Surgical procedures are designed to modify the retropalatal pharynx, the retrolingual pharynx, or both. There is no single surgical procedure, short of tracheostomy, which consistently results in complete elimination of OSAS. However, appropriate application of current surgical techniques (synchronously or sequentially) may achieve cure in most patients without resort to tracheostomy. Patient selection, versatility in varied surgical approaches, and willingness to utilize more than one procedure when necessary appear to be critical attributes of a successful surgical program. On the other hand, analysis of the efficacy of individual surgical interventions is thwarted by the frequent practice of reporting on the application of multiple procedures in combination with evaluation of the composite effect. Well designed, multi center studies would help clarify the strengths and weaknesses of different treatment approaches. PMID- 12531122 TI - The significance of saliva during sleep and the relevance of oromotor movements. AB - Saliva is an essential component of the oroesophageal milieu and allows for normal speech, taste, mastication, food bolus formation and swallowing. Saliva has important functions in protecting the hard and soft tissues of the oral cavity from acids and pathogenic microbes. A large number of people suffer either subjective or objective alterations in quantity and/or quality of their saliva that may be secondary to disease, medications, medical treatments or emotional events. Sleep-related xerostomia is a sensation of dry mouth associated with a report of either mouth and/or throat discomfort that induces awakenings for water intake. The prevalence of self-reported dry mouth complaint during sleep (associated with awakening and water intake) in a Canadian survey was estimated at 23%. The biological significance of decreased saliva during sleep is unknown and it is unclear how the oral cavity compensates for this period of relative dryness. The amount of saliva produced is greatest during the waking hours of the day and diminishes dramatically during sleep and may represent another process in the human body that displays a circadian rhythmicity. Salivary secretion during wakefulness is, in part, associated with oromotor activity involving the masticatory muscles. Rhythmic masticatory muscle activity and swallowing are non disruptive events that occur during normal sleep. We hypothesize herein that lubrication from saliva is necessary during sleep to protect tissue integrity and health of oroesophageal structures. PMID- 12531123 TI - Upper airway muscles awake and asleep. AB - Upper airway (UA) structures are involved in different respiratory and non respiratory tasks. The coordination of agonist and antagonist UA dilators is responsible for their mechanical function and their ability to maintain UA patency throughout the respiratory cycle. The activity of these muscles is linked with central respiratory activity but also depends on UA pressure changes and is greatly influenced by sleep. UA muscles are involved in determining UA resistance and stability (i.e. closing pressure), and the effect of sleep on these variables may be accounted for by its effect on tonic and phasic skeletal muscle activities. The mechanical effects of UA dilator contraction also depend on their physiological properties (capacity to generate tension in vitro, activity of the anaerobic enzymatic pathway, histo-chemical characteristics that may differ between subjects who may or may not have sleep-related obstructive breathing disorders). These characteristics may represent an adaptive process to an increased resistive loading of these muscles. The apparent discrepancy between the occurrence of UA closure and an increased capacity to generate tension in sleep apnea patients may be due to a reduction in the effectiveness of UA muscle contraction in these patients; such an increase in tissue stiffness could be accounted for by peri-muscular tissue characteristics. Therefore, understanding of UA muscle physiological characteristics should take into account its capacity for force production and its mechanical coupling with other UA tissues. Important research goals for the future will be to integrate these issues with other physiological features of the disease, such as UA size and dimension, histological characteristics of UA tissues and the effect of sleep on muscle function. Such integration will better inform understanding of the role of pharyngeal UA muscles in the pathophysiology of the sleep apnea/hypopnea syndrome. PMID- 12531125 TI - Neurobiological bases for the relation between sleep and depression. AB - The serotoninergic system is involved in the regulation of sleep and wakefulness, its activity being at maximum during the awake state and minimum during sleep. In particular, the production of rapid eye movement (REM) sleep depends on the decrease of serotoninergic tone in brain stem structures. Thus, serotoninergic compounds which increase this tone (such as antidepressants) induce inhibition of REM sleep. Depression is associated with a functional decrease of serotoninergic neurotransmission and with specific alterations of sleep, notably insomnia. Paradoxically, even though they complain of sleep loss, depressed patients exhibit significant mood improvement after one night of sleep deprivation. This antidepressant effect can be accounted for by the same serotoninergic mechanisms as those described for pharmacological treatments. Indeed, the therapeutic action of antidepressants such as selective serotonin reuptake inhibitors is thought to depend directly on the enhancement of central serotoninergic neurotransmission. Such enhancement is achieved through desensitization of serotoninergic autoreceptors, which results from chronic treatment with these compounds. Sleep deprivation also induces an activation of serotoninergic neurons due to prolonged wakefulness, and leads to similar serotoninergic adaptive processes. The common neurobiological mechanisms resulting from pharmacological antidepressant treatment and sleep deprivation suggest that sleep loss in some insomniac or in depressed patients might be an endogenous compensatory process which would be therapeutical rather than pathological. This proposal should open the way to new strategies in the treatment of depression. PMID- 12531127 TI - Therapeutic use of sleep deprivation in depression. AB - Total sleep deprivation (TSD) for one whole night improves depressive symptoms in 40-60% of treatments. The degree of clinical change spans a continuum from complete remission to worsening (in 2-7%). Other side effects are sleepiness and (hypo-) mania. Sleep deprivation (SD) response shows up in the SD night or on the following day. Ten to 15% of patients respond after recovery sleep only. After recovery sleep 50-80% of day 1 responders suffer a complete or partial relapse; but improvement can last for weeks. Sleep seems to lead to relapse although this is not necessarily the case. Treatment effects may be stabilised by antidepressant drugs, lithium, shifting of sleep time or light therapy. The best predictor of a therapeutic effect is a large variability of mood. Current opinion is that partial sleep deprivation (PSD) in the second half of the night is equally effective as TSD. There are, however, indications that TSD is superior. Early PSD (i.e. sleeping between 3:00 and 6:00) has the same effect as late PSD given equal sleep duration. New data cast doubt on the time-honoured conviction that REM sleep deprivation is more effective than non-REM SD. Both may work by reducing total sleep time. SD is an unspecific therapy. The main indication is the depressive syndrome. Some studies show positive effects in Parkinson's disease. It is still unknown how sleep deprivation works. PMID- 12531128 TI - Assessment and diagnosis of insomnia in non-pharmacological intervention studies. AB - A number of non-pharmacologic interventions have been developed and studied for the treatment of primary insomnia in adults. Fifty-four non-pharmacological intervention studies published over the last 20 years were reviewed to determine how primary insomnia was assessed and to characterize patients participating in the studies. The main objectives were to inform clinicians who look to the literature for information on the treatment of insomnia about selection and diagnosis of participants, and to examine differences between clinic-based and research-based studies. Mean age for all patients was 47 years; 65% were women. Studies typically used interview procedures to assess participants. The most common exclusion criteria overall was insomnia secondary to medical illness or psychiatric disorder. Methods for assessing these criteria varied widely across studies. Inclusion most commonly required a 6-month duration of insomnia, and sleep difficulty at least 3 nights per week. There were significant differences between clinic- and research-based studies in the screening assessments, exclusion criteria, and participant drop-out rate. The heterogeneity in assessment and diagnosis complicates comparison across studies. Some studies were more likely to include severe insomnia sufferers and/or individuals with secondary insomnia. Comprehensive assessment and complete reporting of research methods is essential to determining the relative efficacy and clinical utility of interventions. Clinicians should be aware of the differences across studies from research vs. clinical settings. To enhance the generalizability of research-based studies to clinical practice, careful screening and description of study participants is essential. When participants are well-described, research-based studies can be useful and informative to clinicians. PMID- 12531129 TI - Bright light, dark and melatonin can promote circadian adaptation in night shift workers. AB - The circadian rhythms of shift workers do not usually phase shift to adapt to working at night and sleeping during the day. This misalignment results in a multitude of negative symptoms including poor performance and reduced alertness during night work and poor daytime sleep at home. After an introduction to circadian principles, we discuss the efficacy of appropriately timed bright light exposure (natural and artificial) and exogenous melatonin administration for producing circadian adaptation to night work. Interventions that generate alternative 24h light/dark patterns that facilitate appropriate circadian phase shifting are discussed. Such interventions include minimizing night workers' exposure to the external light/dark cycle, and the use of intermittent and moving patterns of bright light at work. The efficacy of melatonin in phase shifting circadian rhythms in the field is also addressed and compared to that of bright light. We present sleep/light exposure schedules that could produce circadian adaptation in permanent night workers. We conclude this review by discussing the impact of individual differences on possible circadian interventions and issues associated with the use of bright light interventions in the field. PMID- 12531130 TI - Restless legs syndrome and periodic limb movements of sleep: global therapeutic considerations. PMID- 12531131 TI - Clinical symptomatology and treatment of restless legs syndrome and periodic limb movement disorder. AB - Patients with restless legs syndrome (RLS) suffer from sensory and motor symptoms evoked in the limbs at rest. Symptoms increase in the evening and during the night. The circadian rhythm and the presence of involuntary periodic limb movement in sleep (PLMS) which are frequently associated with arousals probably cause the leading symptom of sleep disturbances in RLS. Patients who do not have typical RLS symptoms but whose polysomnography shows PLMS that either impact on sleep continuity or daytime functioning are diagnosed as having the periodic limb movement disorder (PLMD). Dopaminergic agents such as levodopa/dopa decarboxylase inhibitor (DDI) and dopamine agonists are considered the treatment of choice for RLS and PLMS. This article gives an overview of the epidemiology, pathophysiology, clinical symptomatology and diagnosis of RLS and PLMD and focuses on treatment strategies in both disorders. PMID- 12531132 TI - Awakening from sleep. AB - Awakening is a crucial event for the organism. The transition from sleep to waking implies physiological processes which lead to a new behavioural state. Spontaneous awakenings have varying features which may change as a function of several factors. The latter include intrasleep architecture, circadian phase, time awake, age, or disordered sleep. Despite its clear theoretical and clinical importance, the topic of awakening (in humans) has received little attention so far. This contribution focuses on major issues which relate to awakening from both basic (experimental) and clinical research. Recent knowledge on neurophysiological mechanisms is reported. The experimental data which provide in the human suggestions on the regulation of awakening are discussed, mainly those concerning sleep architecture and homeostatic/circadian factors also in a life span perspective, since age is a powerful factor which may influence awakening. Clinical contributions will examine two main sleep disorders: insomnia and hypersomnia. Daytime functioning is shown in insomniac patients and compared to other pathologies like sleep apnea. A final section evokes links between some types of night waking and psychological factors. PMID- 12531133 TI - Sleepiness in children and adolescents: clinical implications. AB - Over the past decade, excessive sleepiness among children and adolescents has been identified as a major societal concern. Professionals working with pediatric groups must increasingly factor sleepiness into assessments of waking function. We define and discuss excessive sleepiness in children and adolescents and review available evidence regarding effects on behavior, mood, and performance. Findings for daytime sleepiness and subsequent impairment in these domains are classified as robust to unknown. Empirical evidence clearly indicates that children and adolescents experience significant daytime sleepiness as a result of inadequate or disturbed sleep. The specific effect of sleepiness on functional domains in pediatric groups are less well-studied, but existing data suggests that children are likely to experience impairment in behavioral, mood, and performance domains. However, such variables as developmental differences in the type and degree of impairment, the degree of sleep disturbance required to produce impairments, and potential risk and protective factors for the effects of sleepiness in children have yet to be described. Further research is clearly warranted, and we discuss important questions and methodological concerns to encourage inquiry in both clinical and experimental settings. Advice is offered with regard to screening for sleep problems and associated sleepiness with children and adolescents. PMID- 12531134 TI - Mental activity during sleep. AB - Since remote antiquity humankind has believed in the supernatural origin of dreaming. The scientific approach to mental activity during sleep dates back to the eighteenth century. One hundred years ago, psychoanalysis introduced dream analysis for diagnostic and therapeutic purposes. Only 50 years ago psychophysiology made it possible to collect mental sleep activity by means of experimental awakenings while monitoring physiological variables; this approach encouraged investigation into the relationship between the features of sleep activity and sleep state (REM versus NREM). Advances in neurophysiology, in neurochemistry, and recently in brain imaging techniques, have shed light on the roles played by the different cerebral structures in determining specific characteristics of mental activity during sleep. The development of cognitive psychology has enabled investigation of dream generation in terms of output from a complex multilevel system of information processing. In addition to sleep state, other factors, such as the time of the night and the sequence of the NREM REM cycles, have been shown to influence the characteristics of mental activity. The usefulness of investigation of mental sleep activity as a clinical tool is controversial. The psychophysiological approach to mental sleep activity in clinical contexts has enabled the exploration of adaptation processes and contributed to neuropsychological studies on focal and systemic brain pathology. PMID- 12531137 TI - What data do we need to diagnose and treat obstructive sleep apnoea syndrome? PMID- 12531135 TI - Adenosine and sleep. AB - Adenosine is directly linked to the energy metabolism of cells. In the central nervous system an increase in neuronal activity enhances energy consumption as well as extracellular adenosine concentrations. In most brain areas high extracellular adenosine concentrations, through A(1) adenosine receptors, decrease neuronal activity and thus the need for energy. Adenosine seems to act as a direct negative feed-back inhibitor of neuronal activity. Hypoxia and ischemia induce very high extracellular adenosine levels, which may limit further brain damage. In brain areas that regulate cortical vigilance, particularly in the basal forebrain, high extracellular adenosine concentrations, induced by prolonged wakefulness, decrease the activity of presumably cholinergic cells and via this mechanism promote sleep. Our hypothesis is that in the cholinergic basal forebrain prolonged wakefulness induces local energy depletion that generates increases in extracellular adenosine concentrations in this area. In addition to the immediate effects, high extracellular adenosine concentrations also induce intracellular changes in signal transduction and transcription, e.g. increase in A(1) receptor expression and NF-kappaB binding activity. These changes may at least partially mediate the long term effects of prolonged wakefulness. Adenosine may also be a common mediator of the effects of several other sleep-inducing factors. PMID- 12531138 TI - Should all sleep apnoea patients be treated? AB - Sleep apnoea is a condition in which people stop breathing during sleep. A number of studies in general and worker populations have shown that the prevalence of an apnoea-hypopnoea index (AHI) >10 is in the range of 20%. Subjects with an AHI >10 that complain of excessive daytime somnolence, tiredness, asphyxic episodes during the night or non-refreshing sleep, among other symptoms, suffer from the sleep apnoea hypopnoea syndrome (SAHS). The prevalence of SAHS is around 4%. Owing to its high prevalence, clinical symptoms, probable secondary cardiovascular consequences and associated social problems, SAHS has a considerable impact on health, management of which is worth considering. Despite the fact that SAHS treatment has been challenged recent studies conclude that nasal continuous positive airway pressure (nCPAP) is undoubtedly effective in clearly symptomatic patients. Its use in clinical practice is adequately supported in the treatment of moderate to severe SAHS. Further studies are needed in order to define the lower range of symptoms to be treated. One of the most important problems encountered in this area results from the combination of two situations. On the one hand, different epidemiological studies have demonstrated that an AHI >10 without symptoms is present in around 15% of the general population. On the other hand, several studies suggest that having a high AHI, even without secondary symptoms, gives rise to some undesirable effects such as traffic accidents and cardiovascular consequences. In this context, comprehensive epidemiological studies are therefore warranted to define the role of nCPAP treatment especially in those subjects with a high AHI but with few or no symptoms. PMID- 12531139 TI - Should all sleep apnoea patients be treated? Yes. AB - Obstructive sleep apnoea (OSA) is a common condition. Whether an apnoeic patient should be considered for treatment depends on the definition of the syndrome, the rating of the severity and the potential morbidity associated with this condition. We have reviewed several types of evidence that early treatments of OSA deserved. There is a natural evolution of OSA leading to spontaneous aggravation and an increased cardiovascular morbidity in untreated patients in clinical populations. Excessive daytime sleepiness (EDS), the key symptom of the disease, can be found with very low apnoea hypopnoea index (AHI) and eliminated by adequate treatment. Cardiovascular risks are present with very low AHI and there is a high relative risk of developing hypertension in the future when patients present with mild OSA at baseline. The effect of treatment is significant when compared with placebo in the mildest forms of the disease. The clinical benefit, however, is mainly expected in terms of consequences for behavioural morbidity (i.e. reversibility of EDS and its related consequences) as the impact on cardiovascular morbidity is more doubtful and may anyway not be observed on a short-term follow-up. Overall, any OSA syndrome that is clearly responsible for EDS should be considered for treatment. Moreover, there is increasing evidence that the cardiovascular risk has to be taken into account when deciding on treatment, even in asymptomatic patients. All this evidence put together suggests early treatment in OSA. PMID- 12531140 TI - Non-invasive ventilation and sleep. AB - In this paper, we review the effects of nocturnal mechanical ventilation on sleep. Indeed, although non-invasive assisted ventilation during sleep has been applied extensively, the exact effects of this treatment on sleep quality have not been thoroughly studied. In patients with severe chronic obstructive pulmonary disease and severe restrictive ventilatory defects, the resulting respiratory failure is aggravated by the specific effects of sleep on respiration. Non-invasive mechanical ventilation can lead to improvements in both ventilation and sleep quality. However, this is not always the case. Moreover, sleep-related leaks may jeopardize the efficiency of the ventilatory assistance which in turn may result in a deterioration in sleep quality. Non-invasive mechanical ventilation, if applied during sleep, should require a monitoring procedure during sleep with the aim of obtaining the best possible effects both on ventilation and on sleep quality. PMID- 12531141 TI - Circadian rhythm sleep disorders (CRSD). AB - Circadian Rhythm Sleep Disorders (CRSD) are a group of sleep disorders characterized by a malsynchronization between a person's biological clock and the environmental 24-h schedule. These disorders can lead to harmful psychological and functional difficulties and are often misdiagnosed and incorrectly treated due to the fact that doctors are unaware of their existence. In the following review we describe the characteristics of CRSD, diagnosis, treatment as well as their relationship to psychopathology, psychotropic drugs and head trauma. PMID- 12531142 TI - Serotonin and sleep. AB - For 50 years, serotonin has been in the centre of the search for the mechanisms and control of sleep. Serotonergic neurotransmission is related to the behavioural state of the animal and plays an important role in modulation of the behavioural state, by interacting with other brain areas modulating circadian rhythm, sleep and waking. Serotonergic activity may be accompanied by waking or sleep depending on the brain area and receptor type involved in the response, on the current behavioural state and on the concomitant agonism/antagonism of other neurotransmitter systems. PMID- 12531143 TI - Sleep disorders and alcohol. PMID- 12531144 TI - Controversies in the measurement of daytime sleepiness. PMID- 12531145 TI - Daytime sleepiness and its evaluation. AB - Basic models of sleepiness, focusing on the homeostatic and circadian components of sleepiness, are able to predict important fluctuations of sleepiness. However, they fail in explaining certain sleepiness phenomena, as for instance in insomnia patients. To meet this shortcoming, modern models incorporate the arousal component of sleepiness, in addition to the sleep drive. While these models mainly concentrate on short-term changes in sleepiness, "state" sleepiness, there are indications that a stable characteristic level of sleepiness, "trait" sleepiness, is also an important determinant of a person's level of sleepiness. This leads to a conceptualization of sleepiness in which situational factors modify a basal level of sleep drive and arousal. It implies that sleepiness is not a unitary concept and can reflect essentially different states. Multiple sleepiness assessment tools have been proposed in the past. The majority of them offer valuable information, but they do not grasp all aspects of sleepiness. We should bear in mind that tools for assessing sleepiness are always operationalizations reflecting the theoretical framework the investigator has on sleepiness. Hence, rather than searching for a gold standard for the measurement of sleepiness, future research effort should be aimed at linking the various measurement techniques with the hypothesized underlying components of sleepiness on a sound empirical basis. PMID- 12531146 TI - Epidemiology of insomnia: what we know and what we still need to learn. AB - Epidemiologists have published more than 50 studies of insomnia based on data collected in various representative community-dwelling samples or populations. These surveys provide estimates of the prevalence of insomnia according to four definitions: insomnia symptoms, insomnia symptoms with daytime consequences, sleep dissatisfaction and insomnia diagnoses. The first definition, based on insomnia criteria as defined by the DSM-IV, recognizes that about one-third of a general population presents at least one of them. The second definition shows that, when daytime consequences of insomnia are taken into account, the prevalence is between 9% and 15%. The third definition represents 8-18% of the general population. The last definition, more precise and corresponding to a decision-making diagnosis, sets the prevalence at 6% of insomnia diagnoses according to the DSM-IV classification. These four definitions of insomnia have higher prevalence rates in women than in men. The prevalence of insomnia symptoms generally increases with age, while the rates of sleep dissatisfaction and diagnoses have little variation with age. Numerous factors can initiate or maintain insomnia. Mental disorders and organic diseases are the factors that have been the most frequently studied. The association between insomnia and major depressive episodes has been constantly reported: individuals with insomnia are more likely to have a major depressive illness. Longitudinal studies have shown that the persistence of insomnia is associated with the appearance of a new depressive episode. Future epidemiological studies should focus on the natural evolution of insomnia. Epidemiological genetic links of insomnia are yet to be studied. PMID- 12531147 TI - The role of actigraphy in sleep medicine. AB - During the last decade actigraphy (activity-based monitoring) has become an essential tool in sleep research and sleep medicine. The validity, reliability and limitations of actigraphy for documenting sleep-wake patterns have been addressed. Normative data on sleep-wake patterns across development have been collected. Multiple studies have documented the adequacy of actigraphy to distinguish between clinical groups and to identify certain sleep-wake disorders. Actigraphy has also been shown to be effective in documenting the effects of various behavioral and medical interventions on sleep-wake patterns. Actigraphy is less useful for documenting sleep-wake in individuals who have long motionless periods of wakefulness (e.g. insomnia patients) or who have disorders that involve altered motility patterns (e.g. sleep apnea). Potential users should be aware of a number of pitfalls of actigraphy: (1) validity has not been established for all scoring algorithms or devices, or for all clinical groups; (2) actigraphy is not sufficient for diagnosis of sleep disorders in individuals with motor disorders or high motility during sleep; (3) the use of computer scoring algorithms without controlling for potential artifacts can lead to inaccurate and misleading results. PMID- 12531148 TI - Sleep and the hypothalamo-pituitary-adrenocortical system. AB - The intention of this review is to summarize the current knowledge on the bidirectional interaction between sleep EEG and the secretion of corticotropin (ACTH) and cortisol. The administration of various hypothalamic-pituitary- adrenocortical (HPA) hormones and their antagonists exerts specific sleep-EEG changes in several species including humans. It is well documented that corticotropin releasing hormone (CRH) impairs sleep and enhances vigilance. In addition, it may promote REM sleep. Changes in the growth hormone-releasing hormone (GHRH):CRH ratio in favour of CRH appear to contribute to shallow sleep, elevated cortisol levels and blunted GH in depression and ageing. On the other hand, in women GHRH appears to exert CRH-like effects on sleep. Acute cortisol administration increases slow-wave sleep (SWS) and GH, probably due to feedback inhibition of CRH, and inhibits REM sleep. With the mixed glucocorticoid and progesterone receptor antagonist mifepriston sleep is disrupted. Subchronic administration of the glucocorticoid agonist methylprednisolone desinhibited REM sleep. A synergism of elevated CRH and cortisol activity may contribute to REM disinhibition during depression. Also ACTH and vasopressin modulate sleep specifically but their physiological role remains unclear. For example acute icv vasopressin enhances wakefulness in rats, whereas its long-term administration increases SWS in the elderly. In various studies the interaction of sleep EEG and HPA hormones has been investigated at the baseline, after manipulation of sleep wake behaviour and after environmental changes. Most studies agree that the circadian pattern of cortisol is relatively independent from sleep and environmental influences. Some data suggest a major effect of light on cortisol secretion. Sleeping is widely associated with blunting and awakenings are linked with increases of HPA hormones. PMID- 12531149 TI - The functional significance of K-complexes. AB - This paper summarizes the present knowledge about the cellular bases of the sleep K-complex (KC). The KC has two phases: the initial surface-positive wave is due to the synchronous excitation of cortical neurones, while the subsequent surface negative wave represents neuronal hyperpolarization. These variations of membrane potential occur within a slow (<1 Hz) oscillation that characterizes all sleep stages. Therefore, KCs are periodic, and their shape and frequency are modulated by the increasing degree of deafferentation attained by the corticothalamic network with the deepening of the sleep. Within this network, the rhythmic KCs recurring at the frequency of the slow oscillation play a leading role by triggering and grouping other sleep oscillations, such as spindles (7-14 Hz) and delta (1-4 Hz). The KC is mainly a spontaneous event generated in cortical networks. During nocturnal epileptic seizures, the KCs are precursors of paroxysmal spike-wave complexes. PMID- 12531150 TI - REVIEW ARTICLE: Behavioural and cognitive-behavioural interventions for sleep disorders in infants and children: A review. AB - This review covers the literature on behavioural and cognitive-behavioural treatments for sleep disturbance in infants, pre-school, and school-age children. Treatment areas are dyssomnias (disorders of initiating, maintaining, or excessive sleep) and parasomnias (behaviours which occur predominantly during sleep). Interventions aimed at preventing sleep disorder through targeting infant sleep patterns are also examined. Controlled experimental studies are the main focus of this review but case studies and clinical reports are also included. It is concluded that, for families willing to undertake behavioural and cognitive behavioural interventions, some treatments appear effective for some infant and child sleep problems, in the short term at least. The adequacy of current research is discussed, and suggestions for future research are given. PMID- 12531151 TI - Toward a cognitive neuroscience of sleep. PMID- 12531152 TI - Cognitive dysfunction in sleep disorders. AB - Fifty-six studies were reviewed that explored cognitive dysfunctions in people with sleep-related breathing disorders (SRBD, 24 studies), insomnia (18 studies), or narcolepsy (14 studies). Individual study outcomes were grouped according to neuropsychological functions. Available evidence was reviewed separately for SRBD, insomnia and narcolepsy. Consistent evidence was found for impaired driving simulation performance in SRBD patients (92.9% of comparisons with control subjects). Other neuropsychological functions with less pronounced impairment included (i) attention span, divided attention and sustained attention for SRBD patients; (ii) attention span, verbal immediate memory and vigilance for insomniac patients, and (iii) sustained attention, vigilance and driving simulation performance for narcoleptic patients. Reduced performance in tasks measuring attention was found to be higher for SRBD and narcoleptic patients (35.9% and 44.2% of all comparisons, respectively) while this rate was lower for insomniac patients (22.8%). Impairment of memory performance in comparison with control subjects was less pronounced for all three groups, with 20.0% for insomnia, 17.1% for SRBD and 15.6% for narcolepsy. In other areas of cognitive functioning, the data did not allow definite conclusions for any of the patient groups. PMID- 12531153 TI - Cognitive and behavioural performance in children with sleep-related obstructive breathing disorders. AB - Sleep-related obstructive breathing disorders (SROBD) are common in children. While the sequelae of cor pulmonae, and growth and developmental impairment have been well documented, neurocognitive deficits have been less well studied. There is emerging evidence that children with SROBD show reduced neurocognitive functioning especially in the inter-related areas of attentional capacity, memory and cognitive function. In addition, these children show increased problematic behaviour and reduced school performance. Early reports suggest that some of these deficits may be reversible with treatment. The genesis of the defects is unclear but may include hypoxaemia and subtle changes in sleep architecture. The natural history of SROBD and long-term effects on neurocognitive functioning and behaviour remain to be fully examined. PMID- 12531154 TI - Frontal lobe function, sleep loss and fragmented sleep. AB - Recent experimental studies involving total sleep loss, sleep reduction and clinically related sleep fragmentation report impaired performance on tasks of frontal lobe or executive function, including measures of verbal fluency, creativity and planning skills. Severity of sleep disturbance in obstructive sleep apnoea syndrome (OSAS) is correlated with level of executive impairment, with some residual impairment despite treatment (continuous positive airway pressure - CPAP). Executive impairment appears to be more closely related to hypoxaemic events rather than daytime sleepiness. Studies of electroencephalographic (EEG) changes throughout the course of sleep and following sleep deprivation as well as functional neuroimaging and psychophysiological changes (event-related potentials - ERPs) following sleep deprivation provide further indication of the relative importance of the frontal regions of the brain to sleep. However, neurocognitive studies present many inconsistencies, task classification is often ambiguous and, in the absence of any unifying explanation at the level of cognitive mechanisms, the overall picture is one of a disparate range of impairment following sleep loss and sleep fragmentation. Poorly defined concepts of frontal lobe function, executive function, memory and attention, using tasks largely developed with more severe deficit levels in mind, create further difficulties in interpreting current findings. PMID- 12531155 TI - Learning and sleep: the sequential hypothesis. AB - During the last 30 years, paradoxical sleep (PS) has been generally considered as the only type of sleep involved in memory processing, mainly for the consistent increase of PS episodes in laboratory animals learning a relatively complex task, and for the retention deficits induced by post-training PS deprivation. The vicissitudes of this idea, examined in detail by several laboratories, have been critically presented in a number of review articles However, according to a more comprehensive unitary proposal (the sequential hypothesis), memory processing during sleep does require the initial participation of slow-wave sleep (SS) in addition to the subsequent involvement of PS. The evidence supporting this hypothesis, largely derived from experiments concerning rats trained for a two way active avoidance task, is reviewed here in some detail. Recent studies of human sleep are in full agreement with this view. In the rat, the main effect of learning on post-training SS consists in the selective increment in the average duration of SS episodes initiating different types of sleep sequences. Notably, following training for a two-way active avoidance task, the occurrence of this effect in sleep sequences including transition sleep (TS), such as SS-->TS-->W and SS-->TS-->PS, appears related to the processing of memories of the novel avoidance response. Conversely, the occurrence of the same effect in sleep sequences lacking TS may reflect the processing of memories of innate responses (escapes and freezings). Memories of innate and novel responses are assumed to engage in a dynamic competitive interaction to attain control of waking behaviour. Interestingly, in baseline sleep, variables of SS-->TS-->W and SS-->TS ->PS sequences, such as the average duration of SS, TS, and PS episodes, have proved to be good indices of the capacity to learn, as shown by their strong correlations with the number of avoidances scored by rats the following day. Comparable correlations have not been displayed by variables of baseline SS-->W and SS-->PS sequences. PMID- 12531156 TI - Sleep states and memory processes in humans: procedural versus declarative memory systems. AB - The present paper focuses on human studies attempting to relate sleep states to memory processes. These studies typically present learning material to participants and then examine their ability to recall this material after intervening post-training sleep or sleep deprivation. Most experiments utilize either sleep recording or sleep deprivation following task acquisition to reach their conclusions, although cueing and position emission tomography (PET) scan studies have also been done. Results strongly suggest that REM sleep is involved with the efficient memory processing of cognitive procedural material but not declarative material. Although there are some data to suggest that stage 3/4 or NREM sleep is necessary for declarative memory consolidation, NREM may in fact simply be occurring at the same time as another factor that is actually involved in the memory processing. Preliminary results suggest that the length of the NREM REM sleep cycle may be important for declarative memory. Preliminary data also suggest that stage 2 sleep may be involved with the memory for motor procedural but not cognitive procedural tasks. Sleep researchers would do well to capitalize on the latest advancements in memory research by choosing tasks that represent special memory systems and examining their relationships to sleep states. PMID- 12531157 TI - EDITORIAL. Growth, Change and Controversy. PMID- 12531158 TI - Chronic hypnotic use: deadly risks, doubtful benefit. REVIEW ARTICLE. AB - In the United States, roughly 2/3 of all hypnotic prescriptions go to chronic users, who have taken hypnotics for an average of 5 years or more. Two large prospective epidemiological studies have shown that reported hypnotic use, especially use 30 times per month, is associated with an excess hazard of death. Indeed, use of hypnotics 30 times per month is associated with a similar mortality hazard to smoking 1-2 packs of cigarettes per day. Moreover, the hypnotic user's wish to improve daytime function is usually unfulfilled. The preponderance of evidence is that hypnotics impair performance, cognition and memory, increase the risk of automobile accidents and falls and promote unfavourable changes in personality. Due to tolerance, the sleep-promoting effects of hypnotics appear to be lost with chronic use. With long-term use, there is little controlled evidence that hypnotics produce benefits of any sort. More study of long-term hypnotic effects by public agencies is needed, but available evidence weighs strongly against long-term prescribing. PMID- 12531159 TI - Sleep bruxism; an overview of an oromandibular sleep movement disorder. REVIEW ARTICLE. AB - Sleep bruxism (SB) is a stereotyped movement disorder characterized by grinding or clenching of the teeth during sleep. The majority of the population will at some time during their lifetime grind or clench their teeth. It becomes a pathological condition when the subject presents severe tooth damage or complains of non-restorative sleep. The prevalence of SB is difficult to estimate, since quite often the subjects are unaware of having the disorder. There is no gender difference. SB is more frequent in the younger generation, with a decline over age. The symptom recognized in children can persist in adulthood. The aetio pathophysiology is still unclear. SB has been associated with tooth interference, psychosocial and environmental factors, brain transmitters and basal ganglia dysfunction. Attempts have been made to specify the personality traits of bruxers, reported to be greater anxiety or vulnerability to stress; however, this is still controversial. SB subjects were observed to present vigilance-sleepiness and somatic problems. However, they are generally good sleepers. Some authors reported SB during all sleep stages, others observed the majority of bruxe episodes during light sleep and REM and often associated with arousal transients. No abnormalities of the autonomic nervous system could be shown in awake SB subjects. While some studies have shown an association between SB and PLM or breathing disorders, others did not confirm this. There is no specific treatment for SB: each subject has to be individually evaluated and treated. Three management alternatives are used: dental, pharmacological and psychobehavioural. PMID- 12531160 TI - Approach to sleep disorders in the nursing home setting. REVIEW ARTICLE. AB - Evidence suggests that sleep disorders and sleep fragmentation are very common in nursing home residents. A variety of factors contribute to these sleeping difficulties, including age-related changes in sleep; the high prevalence of dementia, depression, medical illness and medications that affect sleep; and sleep disorders such as respiratory disturbance of sleep. Other important factors include common lifestyle characteristics in nursing home residents (such as inactivity, large amounts of time spent in bed, lack of bright light exposure, and poor sleep hygiene) and the disruptive night-time nursing home environment. Recent interventional studies suggest that improvement in the nursing home environment may be an important aspect of the management of sleeping difficulties. Assessment and management of sleeping problems in nursing home residents should involve comprehensive assessment and treatment of the multiple factors that can interfere with sleep. Residents who fail these interventions can be considered for treatment with sleeping medications. Unfortunately, there is little data on the effectiveness of sleeping medications and the specific management of sleep disorders in this setting. Future research should focus on clarifying the contribution of various environmental factors to sleep impairment, and the testing of these various interventions on sleep. PMID- 12531161 TI - Narcolepsy: genetic predisposition and neuropharmacological mechanisms. REVIEW ARTICLE. AB - Narcolepsy is a disabling sleep disorder characterized by excessive daytime somnolence (EDS), cataplexy and REM sleep-related abnormalities. It is a frequently-occurring but under-diagnosed condition that affects 0.02 to 0.18% of the general population in various countries. Although most cases occur sporadically, familial clustering may be observed; the risk of a first-degree relative of a narcoleptic developing narcolepsy is 10-40 times higher than in the general population. The disorder is tightly associated with the specific human leukocyte antigen (HLA) allele, DQB1*0602 [most often in combination with HLA-DR2 (DRB1*15)]. Genetic transmission is, however, likely to be polygenic in most cases, and genetic factors other than HLA-DQ are also likely to be implicated. In addition, environmental factors are involved in disease predisposition; most monozygotic twins pairs reported in the literature are discordant for narcolepsy. Narcolepsy was reported to exist in canines in the early 1970s. Both sporadic and familial cases are also observed in this animal species. A highly-penetrant single autosomal recessive gene, canarc-1, is involved in the transmission of narcolepsy in Doberman pinschers and Labrador retrievers. Positional cloning of this gene is in progress, and a human homologue of this gene, or a gene with a functional relationship to canarc-1, might be involved in some human cases. Human narcolepsy is currently treated with central nervous system (CNS) stimulants for EDS and antidepressants for cataplexy and abnormal REM sleep. These treatments are purely symptomatic and induce numerous side effects. These compounds disturb nocturnal sleep in many patients, and tolerance may develop as a result of continuous treatment. The canine model is an invaluable resource for studying the pharmacological and physiological control of EDS and cataplexy. Experiments using canine narcolepsy have demonstrated that increased cholinergic and decreased monoaminergic transmission are likely to be at the basis of the pathophysiology of the disorder. Pharmacological studies have shown that blockade of norepinephrine uptake mediates the anticataplectic effect of currently prescribed antidepressants, while blockade of dopamine uptake and/or stimulation of dopamine release mediates the awake-promoting effect of CNS stimulants. Studies in canine narcolepsy also suggest that mechanisms and brain sites for triggering cataplexy are not identical to those regulating REM sleep. It may thus be possible to develop new pharmacological compounds that specifically target abnormal symptoms in narcolepsy, but do not disturb physiological sleep/wake cycles. (See also postscript remarks). PMID- 12531162 TI - Origin and Significance of the Cyclic Alternating Pattern (CAP). REVIEW ARTICLE. AB - This article reviews the 15 years of studies conducted on the cyclic alternating pattern (CAP), the electro- encephalogram (EEG) marker of arousal instability during sleep. Starting from the limits and drawbacks of conventional sleep parameters (macrostructure), investigation of the underground world (microstructure) is extensively carried out with particular attention to K complexes and other arousal-related phasic events of non-rapid eye movement (NREM) sleep. The continuity between conventional and non-conventional arousals is stressed in the light of autonomic function and motor activity evidence. The similarities and differences between CAP and arousals are thoroughly illustrated as well as the putative biological bases of these phenomena. The oscillatory nature of CAP, the involvement of CAP in the modulation of EEG synchrony, the sleep-maintenance properties of CAP in response to arousing stimuli, the clinical applications of CAP parameters and the promising availability of automatic scoring systems are highlighted. PMID- 12531163 TI - Computer based sleep recording and analysis. AB - Sleep analysis is based on polysomnography. Modern polysomnographic systems are computer based. Visual and automatic analysis of sleep and respiration is supported by most computer based systems. Four functions can be distinguished in computer based polysomnography: recording, documentation during the recording, automatic and visual analysis and report generation. This review compiles the minimal requirements for digital sleep recording, documentation, analysis and reporting. The basic principles of automatic sleep analysis are reported. The requirements and the basic principles for the analysis of non electroencephalography (EEG) signals, such as respiration, snoring, oxygen saturation, electrocardiography (ECG) and options are reported. New developments in sleep EEG processing are discussed to enlighten how computer based sleep analysis can add quantative parameters to the rules for visual sleep staging established by Rechtschaffen and Kales 30 years ago. This helps to extend our understanding of sleep. PMID- 12531164 TI - Limitations of Rechtschaffen and Kales. AB - The guidelines of Rechtschaffen and Kales (R&K) were meant as a reference method. However, it became, unintentionally, a gold standard. The rules have never been appropriately validated. It is used for the scoring of recordings of, for instance, pathological sleep, for which it was not designed. Today R&K is an insufficient description of sleep processes. In practice, the rules are often difficult or impossible to follow and deviations are common, although not reported. The major drawbacks are: low temporal resolution, ignorance of spatial information, insufficient number of stages, low correspondence between electrophysiological activity and stages, and ignorance of other physiological parameters such as autonomous nervous system activity and body motility. The original idea of the committee should be followed; the guidelines should be revised with the accumulation of new knowledge and development of technology. This is especially true for the analysis of delta waves, which should replace visual scoring of the NREM sleep stages. Many of the problems related to the use of the manual are not due to the system itself but the way it has been applied. This has evidently had a serious impact on the way of thinking and the development of the field in general. PMID- 12531165 TI - Post-traumatic stress disorder and sleep-what a nightmare! AB - According to DSM IV criteria, sleep disturbances are incorporated in the definition of post-traumatic stress disorder (PTSD). These include the re experiencing symptoms (nightmares, criteria B) and a hyperarousal state (difficulty initiating and maintaining sleep, criteria D). PTSD patients commonly complain of sleep disturbances. Moreover, insomnia, restless sleep and trauma related dreams might be the primary complaint of some patients. However, although subjective sleep disturbances are considered characteristic of PTSD, sleep laboratory studies have provided inconsistent evidence of objective sleep disorders. A variety of sleep architectures and sleep patterns has been reported in PTSD. However, only a few studies have controlled for comorbidities. Thus, uncertainty exists to what extent the sustained complaints of sleep disturbances in chronic PTSD are specifically related to the impact of exposure to traumatic stress, or rather are a consequence of comorbid disorders. Specific changes in REM sleep suggest a pathophysiologic role of REM sleep abnormality in PTSD (e.g. anxiety dreams, increased REM density, exaggerated startle response, decreased dream recall and elevated awakening thresholds from REM sleep). However, again, studies have failed to show consistent changes in percentage of REM sleep or in REM latency. There might be a coexistence of pressure to REM along with inhibitory forces of REM that result in high variability of REM parameters across patients. Alternatively, changes in REM sleep might reflect the effect of comorbid psychiatric disorders that results in inconsistent findings between patients. The current review tries to address these issues based on recent studies carried out in this field. PMID- 12531166 TI - The many faces of insomnia. PMID- 12531167 TI - Efficacy and safety of herbal stimulants and sedatives in sleep disorders. AB - World-wide use of herbal medicines is increasing, following regulatory and manufacturing developments. Herbs are attractive alternative medications to many patients with sleep disorders, who may be averse to using conventional drugs. We review here the most common herbal stimulants and sedatives. Caffeine, in herbal teas, black tea, coffee, soft drinks and pharmaceuticals, is used widely to control sleepiness, but more research is needed on its use in sleep disorders. Ephedra, and its constituent ephedrine, are used in both stimulant and weight loss preparations, sometimes with caffeine; safety concerns have arisen with this practice. Yohimbe is another herb used in stimulant and body-building preparations which has safety concerns. Asian and Siberian ginseng have been traditionally used for fatigue, and have some supportive experimental evidence for this use. Herbal sedatives also have some evidence for efficacy; the observations that certain plant flavonoid compounds bind to benzodiazepine receptors adds interest to their use. Valerian and kava have received the most research attention; both have decreased sleep onset time and promoted deeper sleep in small studies, and kava also shows anxiolytic effects. German chamomile, lavender, hops, lemon balm and passionflower are reputed to be mild sedatives but need much more experimental examination. PMID- 12531168 TI - Depression and insomnia: questions of cause and effect. AB - Chronic insomnia is a risk factor for the development of psychiatric disorders, including depression, as well as a prodrome of major depressive episodes, a consequence or complication of depression that often persists beyond the clinical episode, and a prognostic indicator of long-term illness course and treatment response. In addition, sleep is physiologically abnormal in persons at risk for depression; for example, shortened REM sleep latency is present not only during clinical episodes of depression, but also before the clinical episode in subjects at risk for depressive illness. Although insomnia usually disappears as depression is treated, it may persist, indicating heightened vulnerability to depressive relapse or recurrence. Physiological changes in sleep related to depression correlate with the likelihood of response to psychotherapy alone and may also identify which patients are unlikely to do well with psychosocial treatment and, therefore, to need somatic therapy in order to preserve recovery. Electroencephalographic (EEG) sleep changes also correlate with the speed of response and with the brittleness or durability of response (i.eprobability of relapse or recurrence). These observations suggest a close relationship between the regulation of sleep and the regulation of mood. The importance of this relationship is further underscored by recent brain imaging studies of sleep and sleep deprivation in patients with major depression. For example, therapeutic sleep deprivation (TSD) may serve as both a catalyst of rapid antidepressant activity and as a probe of treatment resistance. TSD's effects on brain metabolic rates, especially in limbic areas, may correlate with a therapeutic response to a night of sleep loss and to antidepressant medication. Finally, treating chronic insomnia with newer selective serotonin reuptake inhibitor (SSRI) antidepressant medication may represent an opportunity for preventing complications of insomnia, including depressive illness. PMID- 12531169 TI - Sleep disturbance in generalized anxiety disorder and its treatment. AB - Sleep laboratory and epidemiological studies indicate that insomnia is a frequent finding in patients with psychiatric disorders. In this respect, insomnia associated with a major depression or an anxiety disorder, mainly generalized anxiety disorder (GAD), is the most prevalent diagnosis. According to available evidence, the sleep disturbance associated with mild-to-moderate GAD is a sleep maintenance insomnia, and to a lesser extent a sleep-onset insomnia. Insomnia associated with mild-to-moderate GAD generally responds to psychological treatments and anxiolytic benzodiazepines. Moreover, concomitant administration of hypnotic medication can be contemplated in patients with severe GAD. PMID- 12531170 TI - Insomnia and daytime functioning. AB - Results from the present review challenge the assumption that daytime functioning deficits are associated with insomnia. Objectively-measured daytime sleepiness is not elevated in people with insomnia, and most cognitive/psychomotor tasks do not indicate deficits in people with insomnia. In contrast, a number of studies have found that people with insomnia self-report daytime symptoms such as elevated fatigue, mood disturbance and reduced quality of life. Studies including both objective and subjective daytime measures have generally found agreement between the two types of measures, with both typically indicating a lack of daytime impairment. The present review also summarizes evidence suggesting that reported daytime difficulties are produced by factors other than poor sleep, such as physiological or psychological arousal or sleep need misperception. PMID- 12531171 TI - Behavioural treatments for sleep problems in children and adolescents with physical illness, psychological problems or intellectual disabilities. AB - Young people with physical, psychological or intellectual disabilities or disorders are reported to have more frequent and persistent problems with sleep than their peers without <<<>>>. Sleep disorders affecting the quantity or quality of sleep have effects on a child's daytime functioning and the functioning of their families. Many children with special needs have learning and behaviour problems and their parents (particularly mothers) have increased levels of stress and poorer mental health. This relationship between sleep disorders and learning, and behaviour and family functioning makes it particularly important that children with special needs receive appropriate intervention for their sleep disorders. This may be one way of mitigating these other problems. This review considers the case reports and experimental trials which have used behavioural treatments for sleep problems in children and adolescents with special needs. Behavioural treatments for sleep-wake cycle disorders, sleeplessness, parasomnias and excessive sleepiness are reported. These preliminary reports do suggest that behavioural approaches can be rapidly successful for treating sleep problems, even where the sleep problems are long standing, severe and associated with physical, psychological or intellectual problems. The parent and the clinician should not be deterred from treating the sleep problem in isolation using behavioural treatments. Methodological issues, however, highlight the importance of further and better research. Not all children responded to the behavioural interventions and some needed re implementation of therapy to maintain improvements; the use of heterogeneous groups make the findings and choice of treatment for individuals difficult to interpret. Finally, there are few studies overall, and the majority are case studies rather than controlled studies using multiple baseline designs or randomization and a control group. Careful studies are required in order to establish the relative efficacy of the behavioural techniques and their suitability with homogeneous subgroups of children with special needs. PMID- 12531172 TI - Guest Editorial: Was O. J. sleepwalking? PMID- 12531173 TI - Parasomnias: sleepwalking and the law. AB - A recent, well-publicized case in which murder during sleepwalking was offered as a defense, underscores the fact that sleep medicine specialists are asked to render opinions or judgements regarding culpability in legal cases regarding violence claimed to have arisen from sleepwalking episodes. This review addresses this difficult issue from scientific, clinical and legal aspects, with emphasis upon the need for further research, calling for close collaboration between the legal and medical (both clinical and basic science) professions. PMID- 12531174 TI - Sleep inertia. AB - Sleep inertia is a transitional state of lowered arousal occurring immediately after awakening from sleep and producing a temporary decrement in subsequent performance. Many factors are involved in the characteristics of sleep inertia. The duration of prior sleep can influence the severity of subsequent sleep inertia. Although most studies have focused on sleep inertia after short naps, its effects can be shown after a normal 8-h sleep period. One of the most critical factors is the sleep stage prior to awakening. Abrupt awakening during a slow wave sleep (SWS) episode produces more sleep inertia than awakening in stage 1 or 2, REM sleep being intermediate. Therefore, prior sleep deprivation usually enhances sleep inertia since it increases SWS. There is no direct evidence that sleep inertia exhibits a circadian rhythm. However, it seems that sleep inertia is more intense when awakening occurs near the trough of the core body temperature as compared to its circadian peak. A more controversial issue concerns the time course of sleep inertia. Depending on the studies, it can last from 1 min to 4 h. However, in the absence of major sleep deprivation, the duration of sleep inertia rarely exceeds 30 min. But all these results should be analysed as a function of type of task and dependent variables. Different cognitive functions are probably not sensitive to the same degree to sleep inertia and special attention should be provided to dependent variables as a result of the cognitive processes under review. Finally, sleep disorders represent risk factors which deserve new insight in treatment strategies to counteract the adverse effects of sleep inertia. PMID- 12531175 TI - Sleep-related behavioural adaptations in free-ranging anthropoid primates. AB - Several aspects of behaviour relating to sleep in monkeys and apes are reviewed, including sleeping site selection, approach to and departure from sleeping sites, social behaviour at the sites, and nocturnal activities. Illustrative examples are given for each topic. Good sleeping sites for primates give protection from predators and/or some physical comfort from the elements and other sources of disturbance. Availability of sleeping sites may determine ranging patterns and whether an area is exploited or not. Times of retiring and resumption of daytime activities are influenced by foraging and ranging requirements. Social relationships and their influencing factors continue during the night, including dominance, kinship, affiliation and sex. Social partners may be used for thermoregulation and for increasing postural stability. Primates show a range of solutions to the problems surrounding sleep, and similarities and differences between monkeys and the large-bodied, nest-building great apes are described. Knowledge of natural sleep-related phenomena in non-human primates can provide valuable insights for human sleep research, and vice-versa. PMID- 12531176 TI - The wide clinical spectrum of nocturnal frontal lobe epilepsy. AB - Nocturnal frontal lobe epilepsy (NFLE) has become clinically relevant in recent years. NFLE represents a spectrum of clinical manifestations, ranging from brief, stereotyped, sudden arousals, often recurring several times per night, sometimes with a quasi-periodic pattern, to more complex dystonic-dyskinetic seizures and to prolonged "somnambulic" behaviour. Episodes of increasing intensity have been labelled as paroxysmal arousal (PA), nocturnal paroxysmal dystonia (NPD) and episodic nocturnal wandering (ENW). NFLE affects both sexes with a higher prevalence for men, is frequently cryptogenetic and displays a strong familial trait for parasomnias and epilepsy (NFLE). Seizures appear more frequently between 14 and 20 years of age, but can affect any age and tend to increase in frequency during life. Interictal and ictal scalp electroencephalography (EEG) are often normal, the use of sphenoidal leads may be helpful. Carbamazepine taken at night is often effective at low doses, but a third of the patients are resistant to anti-epileptic drugs (AED) treatment. A familial form, characterized by an autosomal dominant transmission, has also been described. Autosomal dominant nocturnal frontal lobe epilepsy is a genetic variant of NFLE, in itself both clinically and biologically heterogeneous. NFLE should be suspected in the presence of frequent stereotyped paroxysmal nocturnal motor events arising or persisting into adulthood. Videopolysomnography is mandatory to confirm the diagnosis. PMID- 12531177 TI - Exercise and sleep. AB - This paper reviews the literature on the association between exercise and sleep. The epidemiological and experimental evidence for whether or not acute and chronic exercise promote sleep is discussed, as well as moderating factors and agendas for future directions of study. The expectation that exercise will benefit sleep can partly be attributed to traditional hypotheses that sleep serves energy conservation, body restoration or thermoregulatory functions, all of which have guided much of the research in this field. Exercise is a complex activity that can be beneficial to general well-being but may also stress the body. Differences in the exercise protocols studied (e.g. aerobic or anaerobic, intensity, duration) and interactions between individual characteristics (e.g. fitness, age and gender) cloud the current experimental evidence supporting a sleep-enhancing effect of exercise. In addition, the tendency to study changes in small groups of good sleepers may also underestimate the efficacy of exercise for promoting sleep. Athough only moderate effect sizes have been noted, meta analytical techniques have shown that exercise increased total sleep time and delayed REM sleep onset (10 min), increased slow-wave sleep (SWS) and reduced REM sleep (2-5 min). The sleep-promoting efficacy of exercise in normal and clinical populations has yet to be established empirically. PMID- 12531178 TI - HIV mutagenesis and the evolution of antiretroviral drug resistance. AB - The development of antiretroviral drug resistance is a major threat to the effective treatment of human immunodeficiency virus type 1 (HIV-1) infection. Drug treatment failure is associated with accumulation of drug resistance mutations and the evolution of drug resistance. Studies from microbial systems provide evidence for a correlation between drug resistance development and increased pathogen mutation rates. Recent studies with HIV-1 have shown that drugs targeted against reverse transcriptase (RT) as well as drug-resistant RT can increase HIV-1 mutation frequencies. Furthermore, combinations of drug and drug-resistant RT have been found to increase virus mutation frequencies in a multiplicative manner. The correlation of increased HIV-1 mutation rates with the evolution of antiretroviral drug resistance indicates that drug failure could increase the likelihood of further resistance evolving from subsequent drug regimens. PMID- 12531179 TI - Fitness of drug resistant HIV-1: methodology and clinical implications. AB - Recent studies of human immunodeficiency virus type 1 (HIV-1) fitness have examined the potential relationship with plasma viral load, drug resistance, and disease progression. For example, treatment of HIV-1 infected individuals with antiretroviral drugs may result in the selection and emergence of inhibitor resistant variants with reduced replicative capacity. However, it is still unclear whether in vitro HIV-1 fitness has any direct relationship to in vivo disease progression or treatment success. A related question is which in vitro assay of viral fitness is the most appropriate for comparison with in vivo HIV-1 fitness. Characterization of the relative viral fitness of drug-resistant HIV-1 strains may lead to a better understanding of whether or not less fit viruses pose a clinical benefit to the patient. PMID- 12531180 TI - Activation of the PI3K/Akt pathway and chemotherapeutic resistance. AB - The resistance of many types of cancer to conventional chemotherapies is a major factor undermining successful cancer treatment. In this review, the role of a signal transduction pathway comprised of the lipid kinase, phosphatidylinositol 3 kinase (PI3K), and the serine/threonine kinase, Akt (or PKB), in chemotherapeutic resistance will be explored. Activation of this pathway plays a pivotal role in essential cellular functions such as survival, proliferation, migration and differentiation that underlie the biology of human cancer. Akt activation also contributes to tumorigenesis and tumor metastasis, and as shown most recently, resistance to chemotherapy. Modulating Akt activity is now a commonly observed endpoint of chemotherapy administration or administration of chemopreventive agents. Studies performed in vitro and in vivo combining small molecule inhibitors of the PI3K/Akt pathway with standard chemotherapy have been successful in attenuating chemotherapeutic resistance. As a result, small molecules designed to specifically target Akt and other components of the pathway are now being developed for clinical use as single agents and in combination with chemotherapy to overcome therapeutic resistance. Specifically inhibiting Akt activity may be a valid approach to treat cancer and increase the efficacy of chemotherapy. PMID- 12531181 TI - Cell cycle regulatory E3 ubiquitin ligases as anticancer targets. AB - Disregulation of the cell cycle and proliferation play key roles in cellular transformation and tumorigenesis. Such processes are intimately tied to the concentration, localization and activity of enzymes, adapters, receptors, and structural proteins in cells. Ubiquitination of these cellular regulatory proteins, governed by specific enzymes in the ubiquitin (Ub) conjugation cascade, has profound effects on their various functions, most commonly through proteasome targeting and degradation. This review will focus on a variety of E3 Ub ligases as potential oncology drug targets, with particular emphasis on the role of these molecules in the regulation of stability, localization, and activity of key proteins such as tumor suppressors and oncoproteins. E3 ubiquitin ligases that have established roles in cell cycle and apoptosis, such as the anaphase promoting complex (APC), the Skp-1-Cul1-F-box class, and the murine double minute 2 (MDM2) protein, in addition to more recently discovered E3 ubiquitin ligases which may be similarly important in tumorigenesis, (e.g. Smurf family, CHFR, and Efp), will be discussed. We will present evidence to support E3 ligases as good biological targets in the development of anticancer therapeutics and address challenges in drug discovery for these targets. PMID- 12531182 TI - Sterol biosynthesis in Pneumocystis: unique steps that define unique targets. AB - Pneumocystis lacks ergosterol, and several antimycotics that bind ergosterol in fungal membranes or inhibit its synthesis are ineffective against Pneumocystis pneumonia. The organism synthesizes C(28) and C(29) Delta(7) 24-alkylsterols, 24 alkyllanosterol derivatives, and Delta(5) 24-alkylsterols, which may be produced by modifying scavenged Delta(5) sterols. Mammals cannot desaturate C-22 and alkylate C-24 of sterols, thus, these processes are particularly attractive targets for antifungal drug development. Recent data indicate that C-22 desaturation is not, but C-24 alkylation is an attractive target in P. carinii. The P. carinii S-adenosyl-L-methionine:sterol C-24 methyl transferase (SAM:SMT) has unique properties; it prefers lanosterol as its sterol substrate. PMID- 12531183 TI - Calcium influx constitutes the ionic basis for the maintenance of glutamate induced extended neuronal depolarization associated with hippocampal neuronal death. AB - Excessive activation of neuronal glutamate receptors has been implicated in the pathophysiology of stroke, epilepsy, and traumatic brain injury. Previously, it has been demonstrated that excitotoxic glutamate exposure results in the induction of an extended neuronal depolarization (END), as well as protracted elevations in free intracellular calcium ([Ca(2+)](i)). Both END and the prolonged [Ca(2+)](i) elevations were shown to correlate with subsequent neuronal death. In the current study, we used whole-cell current-clamp electrophysiology and fura-ff Ca(2+) imaging to determine the electrophysiological basis of END. We found that removal of extracellular Ca(2+) but not Na(+) in the post-glutamate period resulted in complete reversal of END, allowing neurons to rapidly repolarize to their initial resting membrane potential (RMP). In addition, removal of extracellular Ca(2+) was sufficient to eliminate the protracted [Ca(2+)](i) elevations induced by excitotoxic glutamate exposure. To investigate the mechanism through which extracellular Ca(2+) was effecting these changes, pharmacological antagonists of well-characterized routes of Ca(2+) entry were tested for their effects on END. Antagonists of glutamate receptors and voltage gated Ca(2+) channels (VGCCs) had no significant effect on the membrane potential of neurons in END. Likewise, inhibitors of the Na(+)/Ca(2+) exchange (NCX) were ineffective. In contrast, addition of 500 microM ZnCl(2) or 100 microM GdCl(3) to control extracellular medium (containing normal levels of extracellular Ca(2+)) in the post-glutamate period resulted in rapid and complete reversal of END. Addition of 1mM CdCl(2) to control medium had only modest effects on END. These data provide the first direct evidence that END induced by excitotoxic glutamate exposure is caused by an influx of extracellular Ca(2+) and demonstrate that the previously irreversible condition of END can be reversed by removing extracellular Ca(2+). In addition, understanding the electrophysiological basis of this novel Ca(2+)-induced extended depolarization may provide an insight into the pathophysiology of stroke, traumatic brain injury, and other forms of neuronal injury. PMID- 12531184 TI - Loading neurons with BAPTA-AM activates xbp1 processing indicative of induction of endoplasmic reticulum stress. AB - Loading cells with the calcium chelator BAPTA-AM is an analytical tool which has been used to suppress a rise in cytoplasmic calcium activity under various experimental conditions and thus, to evaluate the role of elevated cytoplasmic calcium levels in the process under investigation. BAPTA-AM may, however, not only have an isolated effect on cytoplasmic processes but also on functions of other subcellular compartments such as the endoplasmic reticulum (ER). Under conditions associated with ER dysfunction, the unfolded protein response is activated which is characterized by suppression of translation and processing of xbp1 mRNA, resulting in activation of the expression of genes coding for ER stress proteins. To investigate whether BAPTA-AM causes ER stress, primary neuronal cell cultures were loaded with varying amounts of BAPTA-AM. Exposure of cells to BAPTA-AM induced a marked rise in processed xbp1 mRNA levels, correlating with exposure times and BAPTA-AM concentrations in the medium used for loading. The increase in processed xbp1 mRNA was associated with suppression of protein synthesis and induction of cell injury. The results of this study indicate that loading primary neuronal cell cultures with BAPTA-AM activates xbp1 processing, implying that this calcium chelator does not have an isolated effect on cytoplasmic calcium activity but also an affect on ER function. PMID- 12531185 TI - pH-dependent effect of mitochondria on calcium influx into Jurkat cells; a novel mechanism of cell protection against calcium entry during energy stress. AB - Loss of the mitochondrial membrane potential results in a significant inhibition of calcium influx through calcium release-activated channels (CRAC) in Jurkat cells suspended in the medium of pH lower than 7.4. This effect disappears when the medium pH increases. Alkalinisation of the cytosol achieved by the addition of NH(4)Cl to the cells pretreated with thapsigargin, CCCP and CaCl(2), suspended in the medium of pH 7.2, does not affect CRAC activity, while alkalisation of the extracellular milieu by NaOH results in a strong stimulation of calcium entry. Thus, the mitochondrial effect on CRAC is exclusively related to the extracellular pH. Coupled mitochondria are able to take up Ca(2+) accumulated in the close proximity of CRAC. This protects these channels against feedback inhibition exerted by high [Ca(2+)](c). We conclude that CRAC may exist in two conformations: inhibitable and not inhibitable by cytosolic Ca(2+). Lower extracellular pH promotes the former one. This explains a much higher inhibitory effect of mitochondrial uncouplers on the calcium influx into the cells exposed to pH 7.2 than that observed in the cells suspended in the medium of pH 7.8. This phenomenon may provide an additional mechanism protecting cells against calcium overloading in transient episodes of energy stress. PMID- 12531186 TI - Actions of ionomycin, 4-BrA23187 and a novel electrogenic Ca2+ ionophore on mitochondria in intact cells. AB - We have used fluorescence digital imaging techniques to explore the actions of two groups of Ca(2+) ionophores: (i). ferutinin, an electrogenic naturally occurring ionophore, and (ii). the neutral ionophores 4-BrA23187 and ionomycin, on cytosolic [Ca(2+)] ([Ca(2+)](c)), mitochondrial [Ca(2+)] ([Ca(2+)](m)) and mitochondrial membrane potential (deltapsi(m)) in HepG2 cells and primary hippocampal neurones in culture. 4-BrA23187 and ionomycin promoted the equilibration of [Ca(2+)] gradients between cellular compartments, including ER, mitochondria and cytosol. Thus, [Ca(2+)](c) and [Ca(2+)](m) increased together and then recovered in parallel on removal of the ionophore. In contrast, following a rise in [Ca(2+)](c) in response to ferutinin, [Ca(2+)](m) remained elevated for prolonged periods after the recovery of [Ca(2+)](c) levels despite washout of the compound. Both groups of Ca(2+) ionophores caused some mitochondrial depolarisation, although this was highly variable in degree. Mitochondrial depolarisation induced by ionomycin and 4-BrA23187 was often modest, independent of cyclosporin A (CsA), was suppressed in the absence of extracellular Ca(2+) and was enhanced by pre-incubation of cells with the inhibitor of the mitochondrial Ca(2+)/2Na(+)-exchanger, CGP37157, suggesting that the change in potential reflects the prior state of mitochondrial calcium loading. The mitochondrial depolarisation induced by ferutinin was not influenced by CGP37157 but was completely blocked by CsA, suggesting that it reflects opening of the mitochondrial permeability transition pore (mPTP). We suggest that ferutinin may provide a very valuable tool to promote mitochondrial calcium overload experimentally and to promote calcium-dependent opening of the mPTP. PMID- 12531187 TI - Ca2+-dependent contractility of isolated and demembranated macronuclei in the hypotrichous ciliate Euplotes aediculatus. AB - The hypotrichous ciliated protozoan Euplotes aediculatus possesses a characteristic C-shaped somatic nucleus (macronucleus) within the cytoplasm, which shows dynamic shape change during the cell cycle. It is shown that isolated macronuclei possess Ca(2+)-dependent contractility. Macronuclei were isolated, stuck fast on the glass surface, and subjected to different concentrations of Ca(2+) in a Ca(2+)-EGTA buffer. The nuclei became expanded at [Ca(2+)]<10(-7)M, and they contracted on subsequent addition of higher concentrations of Ca(2+). Cycles of expansion and contraction of the nucleus could be repeated many times by alternate addition of EGTA and Ca(2+), indicating that the size of isolated nuclei can be regulated by [Ca(2+)] alone. The nuclear contraction was observed in all phases of the cell cycle, but contractility was less evident around replication bands in the S phase. In addition to the hypotrichous ciliate Euplotes, similar Ca(2+)-dependent nuclear contractility was found to exist in other cell types, including protozoans of different taxa (a heliozoon Actinophrys sol and a peniculine ciliate Paramecium bursaria), and also mammalian culture cells (HeLa cells). Our findings suggest a possibility that Ca(2+)-dependent nuclear contractility may be shared among diverse eukaryotic organisms. PMID- 12531188 TI - Ca2+/calmodulin-dependent translocation of sphingosine kinase: role in plasma membrane relocation but not activation. AB - Activation of sphingosine kinase (SPHK), thereby increasing cellular levels of sphingosine 1-phosphate (S1P), may be involved in a variety of intracellular responses including Ca(2+) signaling. This study uses mammalian SPHK1a, tagged with enhanced green fluorescent protein (eGFP), to examine whether translocation of this enzyme is linked with Ca(2+)-mobilizing responses. Real-time confocal imaging of SPHK1a-eGFP in human SH-SY5Y neuroblastoma cells visualized a relocation of the enzyme from the cytosol to the plasma membrane in response to Ca(2+)-mobilizing stimuli (muscarinic M(3)- or lysophosphatidic acid receptor activation, and thapsigargin-mediated store release). This redistribution was preceded by a transient increase in cytosolic SPHK1a-eGFP levels due to liberation of SPHK from localized higher intensity regions. Translocation was dependent on Ca(2+) mobilization from intracellular stores, and was prevented by pretreatment with the Ca(2+)/calmodulin inhibitor W-7, but not W-5 or KN-62. In functional studies, pretreatment with W-7 lowered basal and M(3)-receptor mediated cellular S1P production. However, this pretreatment did not alter agonist-mediated Ca(2+) responses, and SPHK1a-eGFP activity itself appeared insensitive to Ca(2+)/calmodulin and W-7. These data suggest a role for Ca(2+)/calmodulin in controlling the subcellular distribution but not the activity of SPHK1a. PMID- 12531189 TI - Altered calcium regulation in freshly isolated aortic smooth muscle cells from bile duct-ligated rats: role of nitric oxide. AB - In the present study we have analyzed the mechanisms of calcium entry and mobilization in smooth muscle cells (SMCs) freshly isolated from the abdominal aorta of rats with bile duct ligation (BDL). The SMCs were obtained in the day of the experiment after collagenase digestion and loaded with fura-2. The intracellular calcium levels ([Ca](i)) were determined in individual cells by fluorescence microscopy. Baseline [Ca](i) was slightly but significantly lower in SMCs from BDL rats (70.14+/-2.02 nM, n=51) than in controls (80.77+/-3.52, n=44). The application of the purinergic agonists ATP and UTP induced a fast calcium peak and a slow return to baseline. But the calcium responses were significantly smaller in the cells from the BDL rats. Also, the area under the curve (AUC) of the calcium responses elicited by the agonists was always lower in the SMCs from BDL rats as compared to the controls. Similar results were obtained with UTP, but the calcium response of the SMCs from the BDL rats was even lower than that observed with ATP. In experiments performed in the absence of extracellular calcium, both agonists also elevated [Ca](i), although the responses were much smaller than those obtained in the presence of calcium. Again, the peak and AUC responses of the SMCs from BDL rats were significantly lower than those of the controls. Incubation with NNA, a non-specific nitric oxide synthase (NOS) inhibitor, or with NIL, an inducible NOS inhibitor (iNOS), potentiated and normalized the calcium responses of the SMCs obtained from BDL rats. These data indicate that, in SMCs from bile duct-ligated rats, both the entry of calcium and the mobilization from internal stores is defective in response to purinergic agonists. NO, of an inducible origin, is involved in this altered calcium regulation. PMID- 12531190 TI - Plasmodium falciparum malaria parasites display a THG-sensitive Ca2+ pool. PMID- 12531191 TI - Determination of glutathionyl-hemoglobin in human erythrocytes by cation-exchange high-performance liquid chromatography. AB - Since glutathionyl-hemoglobin has been suggested to be a clinical marker of oxidative stress in human blood and given the growing biological relevance of oxidative stress as a pathogenic factor in several diseases, we describe a method to measure glutathionyl-hemoglobin concentration in erythrocytes, by using cation exchange high-pressure liquid chromatography with UV detection. The glutathionyl hemoglobin peak has been identified on the basis of the following findings: (a) the peak increased when the sample was incubated with oxidized glutathione; (b) the peak disappeared when the sample was reduced with dithiothreitol, with the simultaneous increase of that corresponding to hemoglobin A(0); (c) the peak could be detected by incubating hemoglobin A(0) with reduced glutathione; (e) deconvoluted mass spectrum of the glutathionyl-hemoglobin peak showed a 16172.0 Da molecular mass, corresponding to hemoglobin beta bound to glutathione. Glutathionyl-hemoglobin concentration has been determined in erythrocytes of 40 healthy subjects, with a mean value of 2.58+/-0.7%, calculated as the percentage of its peak area ratio to that of total hemoglobin (HbA(0)+HbA(2)+HbA(1C)+glutathionyl-hemoglobin). The availability of a simple and reproducible method to detect glutathionyl-hemoglobin concentration in blood could be useful in monitoring oxidative stress, and for investigating the efficacy of antioxidant therapies in clinical trials. PMID- 12531192 TI - Nucleotide profile of mouse liver: response to 2'-O-dibutyryl cytidene 3',5' cyclic monophosphate. AB - The effect of 2'-O-dibutyryl cytidene 3',5'-cyclic monophosphate (dibutyryl cCMP) on the nucleotide profile of mouse liver was examined. Dibutyryl cCMP caused an increase in the amount of CTP in mouse liver. Perchloric acid extracts of liver tissue were neutralized with tri-N-octylamine in trichlorotriflouroethene and, after removal of CLO(4-), subjected to preliminary purification on a Cu2+-loaded column of Chelex 100. A high-pressure liquid chromatographic anion-exchange procedure was used and gave good resolution of the free nucleotides on a single column. PMID- 12531193 TI - Reduction of autofluorescence on DNA microarrays and slide surfaces by treatment with sodium borohydride. AB - Microarray technology is currently being used extensively in functional genomics research and modern drug discovery and development. Henceforward, tremendous application potential for this technology exists in the fields of clinical diagnostics and prognostics, pathology, and toxicology for high-throughput analysis of "disease" gene expression. However, the major hurdle now in this technology is not the performance of the arrays but rather the efficient reproducibility of the hybridization signal intensity in a fluorescence-based analysis. The sensitivity of fluorescence detection on an array is to a large extent limited by the amount of background signal arising due to nonspecifically bound probes and fluorescence that is intrinsically associated with the chip substrate and/or the attached target DNA, the so-called autofluorescence. Here, we describe a simple and efficient method to reduce autofluorescence from undetermined sources on coated glass slides with and without DNA arrays. This sodium borohydride-mediated reduction process resulted in significantly lower and more even background fluorescence. This in turn extended the dynamic range of detection and reduced the average coefficient of variation of fluorescent signal ratios on DNA microarrays in addition to improving the detection of genes that are expressed at a low level. PMID- 12531194 TI - Simultaneous detection of diverse analytes with an aptazyme ligase array. AB - Allosteric ribozymes (aptazymes) can transduce the noncovalent recognition of analytes into the catalytic generation of readily observable signals. Aptazymes are easily engineered, can detect diverse classes of biologically relevant molecules, and have high signal-to-noise ratios. These features make aptazymes useful candidates for incorporation into biosensor arrays. Allosteric ribozyme ligases that can recognize a variety of analytes ranging from small organics to proteins have been generated. Upon incorporation into an array format, multiple different aptazyme ligases were able to simultaneously detect their cognate analytes with high specificity. Analyte concentrations could be accurately measured into the nanomolar range. The fact that analytes induced the formation of new covalent bonds in aptazyme ligases (as opposed to noncovalent bonds in antibodies) potentiated stringent washing of the array, leading to improved signal-to-noise ratios and limits of detection. PMID- 12531195 TI - Optimizing antibody immobilization strategies for the construction of protein microarrays. AB - Antibody microarrays have the potential to revolutionize protein expression profiling. The intensity of specific signal produced on a feature of such an array is related to the amount of analyte that is captured from the biological mixture by the immobilized antibody (the "capture agent"). This in turn is a function of the surface density and fractional activity of the capture agents. Here we investigate how these two factors are affected by the orientation of the capture agents on the surface. We compare randomly versus specifically oriented capture agents based on both full-sized antibodies and Fab' fragments. Each comparison was performed using three different antibodies and two types of streptavidin-coated monolayer surfaces. The specific orientation of capture agents consistently increases the analyte-binding capacity of the surfaces, with up to 10-fold improvements over surfaces with randomly oriented capture agents. Surface plasmon resonance revealed a dense monolayer of Fab' fragments that are on average 90% active when specifically oriented. Randomly attached Fab's could not be packed at such a high density and generally also had a lower specific activity. These results emphasize the importance of attaching proteins to surfaces such that their binding sites are oriented toward the solution phase. PMID- 12531196 TI - Simultaneous determination of norepinephrine, serotonin, and 5-hydroxyindole-3 acetic acid in microdialysis samples from rat brain by microbore column liquid chromatography with fluorescence detection following derivatization with benzylamine. AB - A microbore column liquid chromatographic method for the simultaneous determination of norepinephrine (NE), serotonin (5-HT), and 5-hydroxyindole-3 acetic acid (5HIAA) in microdialysis samples from rat brain is described. The method is based on precolumn derivatization of NE, 5HT, and 5HIAA with benzylamine in the presence of potassium hexacyanoferrate(III) resulting in the corresponding highly fluorescent and stable benzoxazole derivatives. A 15-microl sample was mixed with 15 microl derivatization reagent solution containing 0.3M 3 cyclohexylaminopropanesulfonic acid buffer (pH 12.0), 0.5M benzylamine, 10mM potassium hexacyanoferrate(III), and methanol (1/1/1/12, v/v/v/v). The derivatization was carried out at 50 degrees C for 20 min. The benzylamine derivatives of NE, 5HT, and 5HIAA were separated on a reversed-phase column (100 x 1.0mm i.d., packed with C18 silica, 5 microm) within 30 min. The mobile phase consisted of 15 mM acetate buffer (pH 5.0) and acetonitrile (31%, v/v); the flow rate was 50 microl/min. The detection limits (signal-to-noise ratio of 3) for NE, 5HT, and 5HIAA in the injection volume of 20 microl were 90, 210, and 260 amol, respectively. Microdialysis samples were collected in 7.5-min intervals from the probes implanted in the hippocampus and prefrontal cortex of awake rats. The basal levels of NE, 5HT, and 5HIAA in the dialysates from the hippocampus were 4.2+/-0.5, 4.9+/-0.6, and 934.1 +/- 63.4 fmol/20 microl, and those from the prefrontal cortex were 6.0+/-1.2,5.51.3, and 669.1 +/- 96.0 fmol/20 microl (mean +/- SE, n=25), respectively. The NE and 5HT levels were altered by perfusion of high-potassium or low-calcium solution and following antidepressant drugs imipramine and desipramine. It is concluded that the new fluorescence derivatization method in combination with microbore column liquid chromatography allows the simultaneous determination of NE, 5HT, and 5HIAA in the microdialysis samples at higher sensitivity, providing easier maintenance in routine use than that achieved by high-performance liquid chromatographic methods with electrochemical detection. PMID- 12531197 TI - Microwave-field-assisted enhanced demixing of aqueous two-phase systems. AB - The slow rate of demixing is a major limitation in wide commercial exploitation of aqueous two-phase systems. In the present work, use of a microwave field has been explored for the first time to enhance phase demixing rates (decrease demixing times) of these systems. The microwave-field-assisted demixing process decreased the demixing time by about 2- to 4-fold in a polyethylene glycol/potassium phosphate system and by about 1.5- to 6.5-fold in a polyethylene glycol/maltodextrin system. The enhanced demixing rate can be explained by the dipole rotation, electrophoretic migration of free salts, multiple reflections at the interfaces, droplet-droplet collision, and reduced viscosity of the continuous phase that occur during the application of a microwave field. PMID- 12531198 TI - Development of intramolecularly quenched fluorescent peptides as substrates of angiotensin-converting enzyme 2. AB - Angiotensin-converting enzyme 2 (ACE2 or ACEH) is a novel angiotensin-converting enzyme-related carboxypeptidase that cleaves a single amino acid from angiotensin I, des-Arg bradykinin, and many other bioactive peptides. Using des-Arg bradykinin as a template, we designed a series of intramolecularly quenched fluorogenic peptide substrates for ACE2. The general structure of the substrates was F-X-Q, in which F was the fluorescent group, Abz, Q was the quenching group (either Phe(NO(2)) or Tyr(NO(2))), and X was the intervening peptide. These substrates were selectively cleaved by recombinant human ACE2, as shown by MS and HPLC. Quenching efficiency increased as the peptide sequence was shortened from 8 to 3 aa, and also when Tyr(NO(2)) was used as a quenching group instead of Phe(NO(2)). Two of the optimized substrates, TBC5180 and TBC5182, produced a signal:noise ratio of better than 20 when hydrolyzed by ACE2. Kinetic measurements with ACE2 were as follows: TBC5180, K(m)=58 microM and k(cat)/K(m)=1.3x10(5)M(-1)s(-1); TBC5182, K(m)=23 microM and k(cat)/K(m)=3.5 x 10(4)M(-1)s(-1). Thus, based on hydrolysis rate, TBC5180 was a better substrate than TBC5182. However, TBC5180 was also hydrolyzed by ACE, whereas TBC5182 was not cleaved, suggesting that TBC5182 was a selective for ACE2. We conclude that these two peptides can be used as fluorescent substrates for high-throughput screening for selective inhibitors of ACE2 enzyme. PMID- 12531199 TI - Identification of the amino acids of human serum albumin involved in the reaction with the naproxen acyl coenzyme A thioester using liquid chromatography combined with fluorescence and mass spectrometric detection. AB - Xenobiotic carboxylic acids, that via their metabolites covalently modify proteins, have been associated with serious side effects in man. Such reactive metabolites may be acyl glucuronides or alternatively, the corresponding acyl-CoA thioesters. In this study, the reaction of a model xenobiotic acyl-CoA, the naproxen-CoA, with human serum albumin (HSA), was characterized by high performance liquid chromatography employing fluorescence and mass spectrometric detection. One mM naproxen-CoA was incubated for 6h with HSA (0.45 mM) at 37 degrees C in a 0.1M phosphate buffer (pH 7.4). The tryptic digest of the reduced and alkylated protein was analyzed in order to identify the amino acids in the sequence that were covalently modified with naproxen. Fluorescent peptides, that represented naproxen-modified peptides, were characterized using HPLC-MS-MS and HPLC-MS in zoom scan mode, which provided information on the structure and the charge of the modified peptides. The naproxen-CoA reacted predominantly with lysine 199, lysine 541, and lysine 351, which was in agreement with the binding pattern that has previously been reported for the reactive acyl glucuronides and their reaction with HSA. PMID- 12531200 TI - Highly sensitive reaction of tryptophan with p-phenylenediamine. AB - A simple and sensitive spectrophotometric method for the determination of tryptophan (TRP) is described. The method is based on the coupling reaction of tryptophan with diazotized p-phenylenediamine dihydrochloride (PPDD) in sulfuric acid medium to give the colored product having an absorption maximum at 520 nm. The coupled product was stable for 2h. Beer's law is obeyed in the tryptophan concentration range of 0.25-11 microg/ml. The method is applied for the analysis of pharmaceutical preparations of tryptophan and also in protein samples for tryptophan. Common excipients used as additives in pharmaceutical preparations do not interfere in the proposed method and the significant feature of the method is that most of the amino acids do not interfere in the determination of tryptophan. PMID- 12531201 TI - Quantitation of CYP1A1 and 1B1 mRNA in polycyclic aromatic hydrocarbon-treated human T-47D and HepG2 cells by a modified bDNA assay using fluorescence detection. AB - The quantitation of mRNA, essential for assessing mechanisms of enzyme regulation, is normally carried out using reverse transcriptase-polymerase chain reaction (RT-PCR). An alternative method uses a signal-amplification nucleic acid probe assay, which measures RNA directly by the QuantiGene Expression Kit and incorporates branched DNA technology from Bayer and luminometer-based readings of a chemilumigenic alkaline phosphatase substrate. To broaden the utility of this assay, we investigated substitution of a fluorescent substrate, 2'-(2 benzothiazol)-6'-hydroxybenzothiazole phosphate and a fluorometer, and applied the method to quantitation of CYP1A1 and 1B1 mRNA in human T-47D and HepG2 cells following induction by benzo[a]pyrene (B[a]P) and dibenzo[a,h]anthracene (DB[a,h]A). The fluorescence response increased linearly for 200 min without photobleaching and increased linearly (r2=0.997) up to at least 0.2 microg total RNA. The data revealed that at 0.5 and 1.0 microM inducing agent, the induction of CYP1A1 mRNA in HepG2 cells by DB[a,h]A exceeded that by B[a]P by 18- and 6 fold, respectively. In T-47D cells B[a]P induced CYP1A1 mRNA by 23-fold and CYP1B1 mRNA by 3.9-fold. A B[a]P cocontaminant in the environment, arsenite, did not affect B[a]P-induced levels of CYP1A1 or 1B1 mRNA in these cells. The modified analytical system provides a rapid-throughput, reproducible, and less labor-intensive method than RT-PCR for quantifying cellular mRNA levels. PMID- 12531202 TI - Fast determination of propranolol in urine and pharmaceutical preparations by stopped-flow and micellar-stabilized room-temperature phosphorescence: validation of the method. AB - The stopped-flow mixing technique has been used to study the kinetic determination of propranolol by means of micellar-stabilized room-temperature phosphorescence. This mixing system diminishes the time required for the deoxygenation of micellar medium by sodium sulfite, allowing a kinetic curve that levels off within only 7s to be obtained. The phosphorescence enhancers thallium (I) nitrate, sodium dodecyl sulfate, and sodium sulfite were optimized to obtain maximum sensitivity and selectivity. A pH value of 6.54 was selected as adequate for phosphorescence development. The kinetic curves of propranolol phosphorescence were scanned at lambda(ex)=290 nm and lambda(em)=524 nm. The calibration graphs were linear for the concentration range from 25 to 400 ng mL( 1). The phosphorescence lifetime of propranolol is approximately 1210 micros. The detection limit calculated as proposed clayton was 13.53 ng mL(-1) and by applying the error propagation theory, the detection limit was 8.37 ng mL(-1). The repeatability was studied using 10 solutions of 200 ng mL(-1) of propranolol; if error propagation theory is assumed, the relative error is 1.94%. The standard deviation for a replicate sample was 4.0 ng mL(-1). This method was successfully applied to the determination of propranolol in commercial formulations and in urine. Suitable recovery values were obtained. PMID- 12531203 TI - Quantitative detection of C-reactive protein using phosphocholine-labelled enzyme or microspheres. AB - C-reactive protein (CRP) is a positive, acute-phase protein. Plasma levels rise dramatically in response to tissue injury or inflammation and fall rapidly after recovery or treatment. Antibody-based human CRP test systems do not readily detect CRP from other animals due to the species specificity of antibodies directed against human CRP. Thus, generic systems for CRP detection, based solely on the interaction between CRP and phosphocholine (PC), have been developed. PC bovine serum albumin (PC-BSA) conjugates were produced and either labeled with horseradish peroxidase to facilitate CRP detection in a CRP enzyme-linked sorbent assay (ELSA) or coupled to carboxy-modified microspheres to facilitate the nonenzymatic, turbidimetric detection of CRP. The CRP-ELSA is a competitive assay, where the total assay time is 45 min, the assay sensitivity is 1.06 mg/L CRP, and the dynamic range of the assay is 0-500 mg/L. When PC-BSA conjugate is covalently coupled to carboxylated microspheres, agglutination occurs in the presence of CRP, the extent of which depends on the quantity of CRP present in the sample. Total assay time is 5 min with a dynamic range of 25-500 mg/L. Both assay formats are capable of accurately detecting human CRP and the CRP-ELSA can detect canine CRP as a disease state indicator. PMID- 12531204 TI - Enhanced nucleic acid capture and flow cytometry detection with peptide nucleic acid probes and tunable-surface microparticles. AB - New methods for automated, direct nucleic acid purification and detection are required for the next generation of unattended environmental monitoring devices. In this study we investigated whether tunable-surface bead chemistry and peptide nucleic acids (PNA) could enhance the recovery and detection of intact rRNA in both test tube and automated suspension array hybridization formats. Intact rRNA was easily captured and detected on PNA-coated Lumavidin beads from 0.1 ng total RNA with a 15-min hybridization in pH 7 buffer, representing 1.7 x 10(3) cell equivalents of total RNA. DNA-conjugated beads in pH 5 hybridization buffer required an overnight hybridization to achieve a detectable signal at 0.1 ng target RNA. Standard DNA hybridization conditions (pH 7) were one order of magnitude less sensitive than the tunable-surface (pH 5) condition. The PNA conjugated particles were 100x more sensitive than the tunable-surface DNA particles in the automated format, with a detection limit of 0.1 ng total RNA. The detection limits for total RNA on PNA-conjugated microparticles is immediately conducive to the detection and characterization of microorganisms in low-biomass environments or to the identification of rare sequences in a complex sample mixture, without using PCR. PMID- 12531205 TI - Lateral-flow and up-converting phosphor reporters to detect single-stranded nucleic acids in a sandwich-hybridization assay. AB - Up-converting Phosphor Technology (UPT) particles were used as reporters in lateral-flow (LF) assays to detect single-stranded nucleic acids. The 400-nm phosphor particles exhibit strong visible luminescence upon excitation with infrared (IR) light resulting in the total absence of background autofluorescence from other biological compounds. A sandwich-type hybridization assay was applied using two sequence-specific oligonucleotides. One of the oligonucleotides probes was covalently bound to the UPT particle (reporter) for direct labeling and detection, whereas the second oligonucleotide probe contained biotin for capture by avidin during LF. The whole procedure of hybridization, UPT-LF detection, and analysis required a minimum time of 20 min. Moreover, aiming at minimal equipment demands, the hybridization conditions were chosen such that the entire assay could be performed at ambient temperature. During lateral flow, only targets hybridized to both capture and detection oligonucleotide were trapped and detected at an avidin capture line on the LF strip. Analysis (IR scanning) of the strips was performed in an adapted microtiter plate reader provided with a 980-nm IR laser for excitation of the phosphor particles (a portable reader was also available). Visible luminescence was measured and presented as relative fluorescence units (RFU) allowing convenient quantitation of the phosphor signal. With the assay described here as little as 0.1 fmol of a specific single-stranded nucleic acid target was detected in a background of 10 microg fish sperm DNA. PMID- 12531206 TI - Simultaneous determination of oxidative hair dye p-phenylenediamine and its metabolites in human and rabbit biological fluids. AB - A liquid chromatography with an electrochemical detector method has been developed for the quantitative measurement for three diamine derivatives (p phenylenediamine, N,N(')-p-phenylenebisacetamide, and 4-aminoacetanilide) in human urine and rabbit blood, urine, and feces. The detection cell consisted of a glassy carbon electrochemical signal obtained with a supporting electrolyte containing 20% methanol-5mM octylammonium orthophosphate (pH 6.30) as the mobile phase. A comparison of the results obtained from HPLC-UV shows agreement. PMID- 12531207 TI - Functional assay to measure yessotoxins in contaminated mussel samples. AB - Yessotoxin (YTX) treatment of MCF-7 cells results in the accumulation of a 100 kDa fragment of E-cadherin (ECRA(100)) without a parallel loss of the intact protein in cytosoluble extracts. As a consequence, concentration-dependent increases in the total immunoreactivity detectable by anti-E-cadherin antibodies relative to controls (RTI) and in the relative immunoreactivity of ECRA(100) (RI) are observed. These responses have been exploited to develop a functional assay to measure YTX in samples from contaminated mussels by a three-step procedure, consisting of (i) treatment of MCF-7 cells with YTX standard in the concentration range 0-1nM and of unknown samples; (ii) preparation of cellular extracts, fractionation of proteins by polyacrylamide gel electrophoresis under denaturing conditions, and immunoblotting with anti-E-cadherin antibodies, followed by densitometric analyses of autoradiographies and calculation of RI of ECRA(100) and of RTI of the samples; and (iii) interpolation of the YTX concentrations in unknown samples on standard curves, by the RI of ECRA(100) and the RTI of the samples. The procedure has been used to measure yessotoxins in contaminated mussel samples, and the results obtained show that this functional assay is very sensitive (limit of detection of about 100ng equivalent YTX/g of digestive gland), and robust, as (i) it is insensitive to matrix effects in the range of toxin concentrations relevant for risk assessment to protect humans from exposure to YTX, (ii) calculations are based on a molecular parameter (the RI of ECRA(100)) which is not affected by errors in sample preparation, (iii) it can be performed by the use of antibodies commercially available from different companies, and (iv) it does not show an absolute need of calibration by a pure standard within each assay. PMID- 12531208 TI - Measurement of oxysterols and alpha-tocopherol in plasma and tissue samples as indices of oxidant stress status. AB - Oxidant stress seems to play a role in several setting of human pathology, such as atherosclerosis, cancer, and aging. The study of oxidant stress in human disease should be based on the evaluation of either sensitive and specific markers of enhanced oxidant stress, such as oxysterols, or antioxidant defense, by measuring alpha-tocopherol. We have developed a rapid method to measure the oxysterols 7beta-hydroxycholesterol and 7-ketocholesterol in plasma (50 healthy subjects) and tissue as an index of oxidant stress in vivo, and from the same sample alpha-tocopherol content. The mean plasma concentration of 7beta hydroxycholesterol and 7-ketocholesterol was 4.6+/-1.1 and 13.4+/-7.6 ng/mL, respectively. Plasma alpha-tocopherol concentration was 5.8+/-1.0 micromol/mol cholesterol. Samples from atherosclerotic plaques contained 20 times more cholesterol, about 45 times higher oxysterols levels, and 600 times more alpha tocopherol compared to normal arteries. No significant difference in cholesterol and oxysterol content was observed between cirrhotic and normal liver. However, cirrhotic liver contained significantly smaller concentration of alpha-tocopherol compared to normal liver. In conclusion, we have developed a rapid and reliable method for the assay of cholesterol oxidation products and alpha-tocopherol in plasma and tissue useful for estimation of oxidant stress/antioxidant balance. PMID- 12531209 TI - Molar absorption coefficients for the reduced Ellman reagent: reassessment. AB - The Ellman method for assaying thiols is based on the reaction of thiols with the chromogenic DTNB (5,5'-dithiobis-2-nitrobenzoate) whereby formation of the yellow dianion of 5-thio-2-nitrobenzoic acid (TNB) is measured. The TNB molar absorption coefficient, 13.6 x 10(3)M(-1)cm(-1), as published by Ellman in 1959 has been almost universally used until now. Over the years, however, slightly different values have been published, and it has further been shown that TNB reveals thermochromic properties. This should be taken into account when the Ellman method is used for determination of enzyme activities, such as in cholinesterase assays. Our data show that the absorbance spectra of TNB are shifted to longer wavelengths when temperature increases, while absorbance maxima decrease. Our recommended molar absorption coefficients at 412 nm are 14.15 x 10(3)M(-1)cm(-1) at 25 degrees C and 13.8 x 10(3)M(-1)cm(-1) at 37 degrees C (0.1M phosphate buffer, pH 7.4). Molar absorption coefficients for other temperatures and wavelengths are included in the paper. PMID- 12531210 TI - Noninvasive evaluation of liver metabolism by 2H and 13C NMR isotopomer analysis of human urine. AB - Mammalian liver disposes of acetaminophen and other ingested xenobiotics by forming soluble glucuronides that are subsequently removed via renal filtration. When given in combination with the stable isotopes 2H and 13C, the glucuronide of acetaminophen isolated from urine provides a convenient "chemical biopsy" for evaluating intermediary metabolism in the liver. Here, we describe isolation and purification of urinary acetaminophen glucuronide and its conversion to monoacetone glucose (MAG). Subsequent 2H and 13C NMR analysis of MAG from normal volunteers after ingestion of 2H2O and [U-13C3]propionate allowed a noninvasive profiling of hepatic gluconeogenic pathways. The method should find use in metabolic studies of infants and other populations where blood sampling is either limited or problematic. PMID- 12531211 TI - Affinity chromatography of DNA on nonporous copolymerized particles of styrene and glycidyl methacrylate with immobilized polynucleotide. AB - Nonporous particles of microsize were prepared by the dispersion polymerization of styrene and glycidyl methacrylate and chemically modified to introduce amino groups on the surface by grafting with either hexamethylenediamine or N-methyl 1,3-propanediamine. Aminated particles were then coupled with phosphorylated single-stranded polynucleotides at the 5'-end through covalent linkages. The affinity columns packed with these prepared polynucleotide-immobilized particles effectively retained single-stranded DNA, which could base-pair with the immobilized sequence. Bound DNAs could be eluted to yield a sharp peak by using an aqueous solution of 0.4M NaOH. The nonspecific adsorption due to the electrostatic interaction between the polynucleotide and the residual amino groups on the particle surface via the amination with hexamethylenediamine was significant and could only be reduced by using a high salt (NaCl) concentration. A higher salt concentration in the elution solution could result in a portion of complementary polynucleotide eluted in the nonretained fraction. However, the nonspecific adsorption of polynucleotides was insignificant in the column packed with DNA-immobilized particles prepared via amination using N-methyl-1,3 propanediamine. The column was effective for microanalysis of sequence-specific DNA. PMID- 12531212 TI - Simultaneous quantification of jasmonic acid and salicylic acid in plants by vapor-phase extraction and gas chromatography-chemical ionization-mass spectrometry. AB - Jasmonic acid and salicylic acid represent important signaling compounds in plant defensive responses against other organisms. Here, we present a new method for the easy, sensitive, and reproducible quantification of both compounds by vapor phase extraction and gas chromatography-positive ion chemical ionization-mass spectrometry. The method is based on a one-step extraction, phase partitioning, methylation with HCl/methanol, and collection of methylated and, thus, volatilized compounds on Super Q filters, thereby omitting further purification steps. Eluted samples are analyzed and quantified by GC/MS with chemical ionization. Standard curves were linear over a range of 5-1000 ng for jasmonic acid and salicylic acid. The correlation coefficients were greater than 0.999 and the recovery rates estimated between 70 and 90% for salicylic acid and 90 and 100% for jasmonic acid. The limit of detection was about 500 fg by using single ion detection mode. Both, cis- and trans-isomers for jasmonic acid can be detected. A comparison with established methods indicates the new method to be highly efficient, allowing reliable quantification of both compounds from small amounts of plant material (5-400mg fresh weight). PMID- 12531213 TI - A real-time assay for DNA sticky-end pairing using molecular beacons. PMID- 12531214 TI - Human whole blood folate analysis using a selected ion monitoring gas chromatography with mass selective detection protocol. PMID- 12531215 TI - Applying malaria parasite's heme detoxification system for screening potential antimalarial drugs. PMID- 12531219 TI - Modeling minimal residual disease (MRD)-testing. AB - There is considerable effort to develop more sensitive methods to detect minimal residual disease (MRD) in bone marrow and blood samples of persons with cancer. Results of MRD-testing are used to predict clinical outcome and determine if more anti-cancer therapy is needed. Mathematical models were developed to assess factors affecting sensitivity and specificity of MRD-testing at diverse cancer cell prevalences. Modeling results and predictions were compared to results of large published studies.Accuracy of MRD-testing depends on cancer cell prevalence and distribution in the blood or bone marrow of the subject, sensitivity and specificity of the MRD-test and sample size. In subjects with low cancer cell prevalences (< or = 10(-4)) results of MRD testing are likely inaccurate. Increasingly sensitive MRD-tests are only marginally useful; the major obstacle to accuracy is inadequate sampling. Increasing sensitivity of methods to detect MRD is unlikely sufficient to increase accuracy of MRD-testing. In contrast, increased sampling (size and frequency) and assigning a high cut-off value (for example, > or = 10(-3)) to declare a MRD-test positive will increase sensitivity and specificity, respectively. PMID- 12531218 TI - Antigen-driven clonal T cell expansion in disorders of hematopoiesis. AB - Patients with myelodysplastic syndrome, aplastic anemia, and LGL leukemia may have overlapping clinical features. This commentary highlights a possible common pathogenetic mechanism of antigen-driven cytotoxic T lymphocytes mediating hematopoietic failure. PMID- 12531220 TI - Mitoxantrone and prolonged infusion gemcitabine as salvage therapy in patients with acute myelogenous leukemia. AB - In a recent phase I study, a combination of gemcitabine at 10 mg/(m(2)min) for 12 h and mitoxantrone 12 mg/m(2) daily for 3 days, achieved a complete remission (CR) in 3 of 12 (25%) patients with refractory leukemia. A pilot assessment of this regimen was conducted to determine its tolerability in patients with refractory acute myeloid leukemia (AML). In a cohort of 18 patients with very refractory disease (6 primary refractory, 12 relapsed, mean initial CR duration 3.5 months), one patient achieved a CR, a second CR with incomplete platelet recovery (CRp). Sepsis and mucositis were significant extramedullary toxicities. The gemcitabine and mitoxantrone regimen was feasible to administer even in heavily pre-treated patients. It was not active in patients who had failed a prior salvage regimen. It may warrant further study in patients with primary resistant AML. PMID- 12531221 TI - Determination of T-cell receptors of clonal CD8-positive T-cells in myelodysplastic syndrome with erythroid hypoplasia. AB - We determined T-cell receptor alpha-chain variable (TCRAV) and T-cell receptor beta-chain variable (TCRBV) region repertoires in peripheral bloods from patients with myelodysplastic syndrome (MDS) with erythroid hypoplasia. T-cells bearing TCR ADV14S1/BV5S2, AV21S1/BV21S4, and AV2S2/BV7S2 segments were markedly increased in three of four MDS patients, respectively. In addition, there was a positive relationship between the increase in the number of CD8-positive T-cells and the expression levels of these TCR transcripts. These findings suggest that CD8-positive T-cells monoclonally or oligoclonally expanded in the peripheral blood. We also determined the nucleotide and amino acid sequences of the complementarity-determining region 3 (CDR3) of TCR alpha- and beta-chains of the expanded T-cells. Unique sequences were detected in a high percentage of the respective CDR3 clones. The gene segment of the variable and joining regions, however, varied among the patients. The deduced amino acid sequences of CDR3 were heterogeneous among the patients, and there was no common motif. These results indicate there is monoclonal or oligoclonal proliferation of CD8-positive T-cells in MDS patients with erythroid hypoplasia, and suggest that these proliferating T cells are responsible for the pathogenesis of the MDS entity. PMID- 12531222 TI - Timed sequential therapy of acute myelogenous leukemia in adults: a phase II study of retinoids in combination with the sequential administration of cytosine arabinoside, idarubicin and etoposide. AB - Clinical outcome in acute myeloid leukemia (AML) is unsatisfactory. One strategy to augment cytotoxicity is TST. All-trans retinoic acid (ATRA) down-regulates bcl 2 expression and heightens AML sensitivity to cytosine arabinoside (ara-C) induced apoptosis in vitro. We designed a trial of ATRA plus ara-C-based TST in an attempt to enhance drug-induced apoptosis and clinical outcome. Between January 1998 and February 2000, 63 patients received induction TST (oral ATRA days 1-6, ara-C and idarubicin days 2-4, VP-16 days 9-11) followed by consolidation TST (ATRA, ara-C and idarubicin followed by a second ara-C infusion days 11-13). Complete remission (CR) was 60%, with higher rates for patients of <60 years (79%), de novo AML (70%), and non-adverse cytogenetics (81%). Median disease-free survival (DFS) for CR patients was 11.2 months (32% at 3+ years). For patients <60 years with de novo AML and non-adverse cytogenetics who underwent two-cycle TST, DFS was 67% at 3+ years. However, patients of age equal to 60 years and those with poor-risk disease features still have poor CR and DFS, despite the addition of ATRA. PMID- 12531223 TI - A phase I/II study of the MDR modulator Valspodar (PSC 833) combined with daunorubicin and cytarabine in patients with relapsed and primary refractory acute myeloid leukemia. AB - The cyclosporine analog Valspodar (PSC 833, Novartis Pharma) is a strong inhibitor of the mdr1 gene product p-glycoprotein (pgp). A phase I/II study was conducted in order to evaluate if addition of Valspodar to treatment with daunorubicin and cytarabine, given to patients with primary refractory or relapsed acute myeloid leukemia, could increase the complete remission rate.Fifty three patients were treated in cohorts of three to six patients. Twelve patients reached a complete remission in bone marrow, five of whom also normalized their peripheral blood values. Three patients experienced treatment-related deaths from pneumonia, liver failure and cerebral hemorrhage, respectively. It is concluded that Valspodar 10 mg/kg per 24 h in combination with daunorubicin 45 mg/m(2) for 3 days and cytarabine 1 g/m(2) twice daily for 4 days is tolerable in this heavily pre-treated group of patients. Due to the moderate treatment results, the phase II part of the study was ended prematurely. The modulation of only pgp did not give an obvious improvement of the treatment results in this group of patients. PMID- 12531224 TI - Prolonged adenovirus-mediated expression of p27(Kip1) unveils unexpected effects of this protein on the phenotype of SUDHL-1 cells derived from t(2;5)-anaplastic large cell lymphoma. AB - SUDHL-1 cells, derived from human t(2;5)-anaplastic large cell lymphoma (ALCL) were kept in culture for 25 days and analyzed for p27(Kip1)-expression, cell cycle, apoptosis, morphology and phenotype as compared with matched controls at different time points after infection with recombinant adenovirus expressing p27(Kip1) (Adp27). The presence of any change in the cell phenotype occurring during the persistent exogenous expression of p27(Kip1) would be indicative of a "clone" of cells surviving apoptosis by reversing G1 arrest. The level of alpha(nu)beta(5) integrin was completely down regulated in cells infected with Adp27 as assessed by flow cytometry and by immunoprecipitation analysis after 14 days of infection. SUDHL-1 cells regained a significant level of alpha(nu)beta(5) integrin on the cell surface after 25 days of infection with Adp27. Based on the importance of integrins in tumor cell proliferation, we speculate that the down regulation of alpha(nu)beta(5) integrin on the surface of SUDHL-1 cells may represent a less tumorigenic phenotype occurring as a consequence of prolonged expression of p27(Kip1). PMID- 12531225 TI - Ex vivo drug and irradiation sensitivities in hypermutated and unmutated forms of chronic lymphocytic leukemia cells. AB - Several investigators have now established that chronic lymphocytic leukemia (CLL) is not a uniform disease entity, since approximately half of the cases of CLL have undergone immunoglobulin V region (IgV) hypermutation, whereas the other half display unmutated Ig genes. The median survival time of mutated CLL (M-CLL) cases has been shown to be approximately twice as long as that for unmutated CLL (UM-CLL), but no clear explanation for this difference is currently available. In this work, we have investigated a cohort of previously untreated CLL patients, to see whether the ex vivo sensitivities of leukemic cells of 16 UM-CLL patients differ from those of 8 M-CLL patients, using nine different drugs and two types of irradiation. Our results demonstrated very similar ex vivo sensitivities and tumor cell heterogeneity of sensitivity of UM-CLL and M-CLL cells when tested against chlorambucil, 2-chloro-2'-deoxyadenosine, cyclosporin A, cis platinum(II)diammine-dichloride, doxorubicin hydrochloride, 2-fluoroadenine-9 beta-D-arabinofuranoside, prednisolone sodium succinate, verapamil, vincristine, gamma-irradiation, and UV-irradiation. This indicates that de novo chemo/radiosensitivity cannot explain the survival difference observed between UM CLL and M-CLL. PMID- 12531226 TI - Humanized anti-interleukin-6 receptor monoclonal antibody induced apoptosis of fresh and cloned human myeloma cells in vitro. AB - We investigated the effect of anti-IL-6 receptor monoclonal antibody (hPM1) on the in vitro proliferation of cloned and freshly isolated myeloma cells from 20 patients with advanced stage multiple myeloma (MM). Humanized PM1 significantly inhibited the growth of a myeloma cell line in a dose-dependent manner and inhibited more than 30% of the proliferation of fresh myeloma cells in 10 of the 19 cases. Flow cytometric analysis using annexin V and 7AAD showed that hPM1 induced apoptosis of myeloma cells. These observations suggest the possibility of using hPM1 for treating some patients with MM whose growth depends on IL-6. PMID- 12531227 TI - BALB/C mice injected with LSTRA leukemic cell line are cured by in vivo treatment with IL-2 + GM-CSF. AB - We studied the effects of the in vivo administration of interleukin-2 (IL-2) (at low doses) + GM-CSF in BALB/C mice injected intraperitoneally with LSTRA murine leukemic cell line in order to test the possible role of this immunotherapeutic approach in the eradication of leukemia. Mice were injected intraperitoneally on day -1 with different concentrations of LSTRA cells. On day 0, mice received a lethal dose of TBI of 700 cGy from a Cs(137) source, followed by a single high dose of recombinant human-granulocyte-colony stimulating factor (rh-G-CSF) (1 microg/g) intraperitoneally 2h after TBI. This procedure rescued 80% of mice but only mice injected with the lower concentration of LSTRA cells (10(3)) could be cured. The lethal dose (LD 100/60) of LSTRA in normal mice was 10(4). The subcutaneous administration of rh-IL-2 (4.000 IU/mouse per day, from day +1 to +28) + rm-GM-CSF (1 microg/kg per day) from day +7 to +28 cured mice injected with 10(4) LSTRA cells and 40% of mice injected with 10(5) cells. Mice injected with 10(6) cells died from leukemia. We observed an increase in LAK activity on days +21 and +28 without an increase in NK activity. We show that BALB/C mice injected with LSTRA cell line can be cured by in vivo activation with IL-2 + GM CSF depending on the cell dose injected. The curability of leukemia was confirmed at the molecular level with a PCR method. PMID- 12531228 TI - Frameshift mutations in caspase-5 and other target genes in leukemia and lymphoma cell lines having microsatellite instability. AB - Microsatellite instability (MSI) was examined in 49 lymphoid leukemia/lymphoma cell lines. Frameshift mutations within eight MSI target genes were analyzed in the 12 cell lines and nine childhood acute lymphoblastic leukemia (ALL) samples with MSI. Six cell lines (50%) showed mutations within the coding repeats of the BLM, caspase-5, TCF-4, and PTEN genes. No mutations were identified in the nine childhood ALL samples with MSI. This is the first report that describes mutations of these genes in hematological malignancies. Mutations, presumably caused by an abnormality of the DNA repair genes, may be selected for during the establishment of cell lines. PMID- 12531229 TI - t(4;12)(q11;p13): a rare chromosomal translocation in acute myeloid leukemia. AB - We present a case of acute myeloid leukemia with t(4;12). This translocation is rare and has been observed in acute leukemias with different but immature phenotypes. To the best of our knowledge, there are around 15 descriptions of t(4;12) in AML, and most interesting, presenting morphological aspects of a pseudo-lymphoid cell with dysplasia of other series. PMID- 12531230 TI - Ph-negative acute lymphocytic leukemia occurring after interferon therapy for Ph positive chronic myelocytic leukemia. AB - We report a unique case of chronic myelocytic leukemia (CML) with the Philadelphia (Ph) chromosome. The patient obtained cytogenetic complete remission (CR) after treatment with interferon (IFN). When he transformed to acute lymphocytic leukemia (ALL), cytogenetic analysis showed that the karyotype was normal and fluorescence in situ hybridization (FISH) indicated that blast cells were Ph-negative. Immunophenotyping showed that the cells were CD34-positive and CD38-negative. Blast cells in his final stage were BCR/ABL rearrangement negative by reverse transcription polymerase chain reaction (RT-PCR). PMID- 12531231 TI - B lineage acute lymphoblastic leukemia transformation in a child with juvenile myelomonocytic leukemia, type 1 neurofibromatosis and monosomy of chromosome 7. Possible implications in the leukemogenesis. AB - This report describes the case of an 8-month-old infant with a diagnosis of juvenile myelomonocytic leukemia (JMML) and type 1 neurofibromatosis that presented progression to B lineage acute lymphoid leukemia (ALL). The same rearrangement of gene T-cell receptor gamma (TCR gamma) was detected upon diagnosis of JMML and ALL, suggesting that both neoplasias may have evolved from the same clone. Our results support the theory that JMML may derive from pluripotential cells and that the occurrence of monosomy of chromosome 7 within a clone of cells having an aberrant neurofibromatosis type 1 (NF1) gene may be the cause of JMML and acute leukemia. PMID- 12531232 TI - Junctional complexes of the blood-brain barrier: permeability changes in neuroinflammation. AB - A wide range of central nervous system (CNS) disorders include neuroinflammatory events that perturb blood-brain barrier (BBB) integrity. Mechanisms by which the BBB responds to physiological and pathological stimuli involve signaling systems in the tight and adherens junctions of the cerebral endothelium. In this review, we examine the molecular composition and regulatory mediators that control BBB permeability and assess how these mediators may be dysregulated in stroke, multiple sclerosis, brain tumors, and meningioencephalitis. An understanding of these molecular substrates in BBB regulation may lead to new approaches for enhancing CNS drug delivery and ameliorating brain edema after injury and inflammation. PMID- 12531233 TI - An inflammatory review of Parkinson's disease. AB - The symptoms of Parkinson's disease (PD) were first described nearly two centuries ago and its characteristic pathology identified nearly a century ago, yet its pathogenesis is still poorly understood. Parkinson's disease is the most prevalent neurodegenerative movement disorder and research into its pathogenesis recently accelerated following the identification of a number of causal genetic mutations. The mutant gene products all cause dysfunction of the ubiquitin proteosome system, identifying protein modification and degradation as critical for pathogenesis. Modified non-degraded intracellular proteins accumulate in certain neuronal populations in all forms of the disease. However, neuronal degeneration is more highly selective and associates with substantial activation of microglia, the inflammatory cells of the brain. We review the current change in thinking regarding the role of microglia in the brain in the context of Parkinson's disease and animal models of the disease. Comparison of the cellular tissue changes across a number of animal models using diverse stimuli to mimic Parkinson's disease reveals a consistent pattern implicating microglia as the effector for the selective degeneration of dopaminergic neurons. While previous reviews have concentrated on the intracellular neuronal changes in Parkinson's disease, we highlight the cell to cell interactions and immune regulation critical for neuronal homeostasis and survival in Parkinson's disease. PMID- 12531234 TI - The role of neuronal nicotinic acetylcholine receptor subunits in autonomic ganglia: lessons from knockout mice. AB - Neuronal nicotinic acetylcholine receptors (nAChR), composed of 12 subunits (alpha2-alpha10, beta2-beta4), are expressed in autonomic ganglia, playing a central role in autonomic transmission. The repertoire of nicotinic subunits in autonomic ganglia includes alpha3, alpha5, alpha7, beta2 and beta4 subunits. In the last 10 years, heterologous expression studies have revealed much about the nature of neuronal nAChRs. However, there is only limited understanding of subunit actions in autonomic system. Functional deletions of subunit by gene knockout in animals could overcome these limitations. We review recent studies on nAChRs on autonomic ganglia for physiological and pharmacological properties and potential locations of the subunits. PMID- 12531235 TI - Remyelinating strategies for the treatment of multiple sclerosis. AB - Demyelination is the pathological hallmark of multiple sclerosis (MS) lesions. The concept of remyelination has gained acceptance in recent years, but naturally occurring remyelination is incomplete. To improve repair processes, a number of strategies have been explored experimentally and clinical trials are being carried out. In principle, remyelination can be achieved by either promoting endogenous repair mechanisms or by providing an exogenous source of myelinating cells via transplantation. Both approaches have been successful in animal models of demyelination. Besides, many studies have elucidated principal mechanisms of oligodendrocyte biology and remyelination in the central nervous system (CNS). This progress in knowledge also allowed for more specific interventions. First clinical trials to enhance endogenous remyelination have been performed, unfortunately with disappointingly negative results. This illustrates that experimental data cannot be easily transferred to human disease, and more detailed knowledge on the regulatory mechanisms of remyelination in MS is required. Recently, the first MS patient received a transplant of autologous Schwann cells. Many other cell types are being studied experimentally, including stem cells. Despite the ethical problems associated with an embryonic cell source, new developments in stem cell biology indicate that adult stem cells or bone marrow-derived cells may substitute for embryonic cells in the future. In this review, we describe the current views on oligodendrocyte biology, myelination and remyelination, and focus on recent developments leading to reconstructing, remyelinating strategies in MS. PMID- 12531236 TI - The hepatocyte: heterogeneity and plasticity of liver cells. AB - The heterogeneity of the proliferative response of the liver to injury and during hepatocarcinogenesis implicates at least four levels of cells in the hepatic lineage which respond: the mature unipolar hepatocyte, the unipolar duct cell, a bipolar ductular progenitor cell, and a multipotent periductular liver progenitor cell of presumed extra-hepatic (bone marrow) origin (1). The original concept of "embryonal rests" as the cells or origin of cancer in adult organs (2) may be correct in the sense that progenitor cells in adult tissues that give rise to cancers may derive from circulating pluripotent stem cells. PMID- 12531237 TI - Airway smooth muscle: contraction and beyond. AB - Airway smooth muscle (ASM), an important tissue involved in the regulation of bronchomotor tone, exists in the trachea and in the bronchial tree up to the terminal bronchioles. The physiological relevance of ASM in healthy airways remains unclear. Evidence, however, suggests that ASM undergoes marked phenotypic modulation in lung development and in disease states such as asthma, chronic bronchitis and emphysema. The shortening of ASM regulates airway luminal diameter and modulates airway resistance, which can be augmented by cytokines as well as extracellular matrix alterations. ASM may also serve immunomodulatory functions, which are mediated by the secretion of pro-inflammatory mediators such as cytokines and chemokines. In addition, ASM mass increases in chronic airway diseases and may represent either a pathologic or an injury-repair response due to chronic inflammation. This review will present evidence that ASM, a "passive" contractile tissue, may become an "active participant" in modulating inflammation in chronic lung diseases. Cell facts 1. Found in the trachea and along the bronchial tree. 2. Critically important in regulating bronchomotor tone of the airways. 3. Differentiation state is associated with the expression of various "contractile proteins." 4. Displays phenotypic modulation of mechanical, synthetic and proliferative responses. 5. Secretes cytokines, chemokines and extracellular matrix proteins. 6. May serve as a potential new target for the treatment of chronic lung diseases. PMID- 12531238 TI - Anti-neutrophil cytoplasm antibody associated vasculitis. AB - Anti-neutrophil cytoplasm antibody associated vasculitides are autoimmune diseases that require treatment with powerful immunosuppressants in order to avert life threatening complications, principally renal failure and lung haemorrhage. Their pathogenesis remains unclear but the circulating autoantibodies are almost certainly instrumental in mediating vascular inflammation. Recent insights into leukocyte-autoantibody interactions support this assertion and may direct future therapeutic strategies, perhaps circumventing some of the toxicity associated with currently used drugs. PMID- 12531239 TI - Transplacental transmission of HIV: a potential role for HIV binding lectins. AB - Although the majority of vertical transmission of HIV occurs around the time of birth, 1.5-2% of pregnancies in HIV-positive women appear to result in the vertical transmission of HIV across the placenta. HIV infection of a number of placental cell types has been demonstrated, but the exact mechanisms of intrauterine vertical transmission remain obscure. The recent discovery of the HIV binding lectins dendritic cell-specific ICAM-grabbing non-integrin (DC-SIGN) and DC-SIGN-related molecule (DC-SIGNR) provides one possible explanation. Cells expressing these lectins are able to adsorb the virus and mediate high efficiency HIV infection of other cell types. Both lectins are expressed by the placenta, with DC-SIGN expression also being present on maternal cells intimately associated with the placenta. This review focuses on possible mechanisms by which these lectins may potentiate the intrauterine vertical transmission of HIV. PMID- 12531240 TI - Ctr1 and its role in body copper homeostasis. AB - Members of the Cu transporter (Ctr) family have been reported to be part of the copper uptake machinery in several organisms. Recently it has been suggested that human Ctr1 (hCtr1) may act as a copper transporter in several tissues including the intestine. hCtr1 is a 190 amino acid protein and is predicted to have three transmembrane-spanning domains and exist in the plasma membrane as a homo-trimer. Ctr1-transfected cell lines exhibit saturable, pH-dependent Cu(I) uptake indicating a role in copper transport. Recent studies with Ctr1 knockout mice have highlighted an essential function in mammalian embryonic development since homozygous mutants die in utero. Heterozygotes are indistinguishable from wild type littermates but have a severely reduced brain copper content, suggesting that Ctr1 is a key component of the copper uptake pathway in the brain. However, its role in other tissues remains elusive. PMID- 12531241 TI - Transferrin receptor 2: a new molecule in iron metabolism. AB - Transferrin receptor 1 (TfR1) which mediates uptake of transferrin-bound iron, is essential for life in mammals. Recently, a close homologue of human transferrin receptor 1 was cloned and called transferrin receptor 2 (TfR2). A similar molecule has been identified in the mouse. Human transferrin receptor 2 is 45% identical with transferrin receptor 1 in the extracellular domain, but contains no iron responsive element in its mRNA and is apparently not regulated by intracellular iron concentration nor by interaction with HFE. Transferrin receptor 2, like transferrin receptor 1, binds transferrin in a pH-dependent manner (but with 25 times lower affinity) and delivers iron to cells. However, transferrin receptor 2 distribution differs from transferrin receptor 1, increasing in differentiating hepatocytes and decreasing in differentiating erythroblasts. Expression of both receptors is cell cycle dependent. Mutations in the human transferrin receptor 2 gene cause iron overload disease, suggesting it has a role in iron homeostasis. PMID- 12531242 TI - Mammalian protein histidine kinases. AB - The existence of protein kinases, known as histidine kinases, which phosphorylate their substrates on histidine residues has been well documented in bacteria and also in lower eukaryotes such as yeast and plants. Their biological roles in cellular signalling pathways within these organisms have also been well characterised. The evidence for the existence of such enzymes in mammalian cells is much less well established and little has been determined about their cellular functions. The aim of the current review is to present a summary of what is known about mammalian histidine kinases. In addition, by consideration of the chemistry of phosphohistidine, what is currently known of some mammalian histidine kinases and the way in which they act in bacteria and other eukaryotes, a general role for mammalian histidine kinases is proposed. A histidine kinase phosphorylates a substrate protein, by virtue of the relatively high free energy of hydrolysis of phosphohistidine the phosphate group is easily transferred to either a small molecule or another protein with which the phosphorylated substrate protein specifically interacts. This allows a signalling process to occur, which may be downregulated by the action of phosphatases. Given the known importance of protein phosphorylation to the regulation of almost all aspects of cellular function, the investigation of the largely unexplored area of histidine phosphorylation in mammalian cells is likely to provide a greater understanding of cellular action and possibly provide a new set of therapeutic drug targets. PMID- 12531243 TI - Differential hypersensitivity to DNase I in the regulatory region of human hsp90 beta gene in heat shock and constitutive expression. AB - DNase I hypersensitivity analysis is a useful tool to investigate impact of structure changes in chromatin on the expression of a gene. In order to unravel chromatin regulation on human hsp90 beta gene, differential sensitivity to DNase I in non-treated and heat-shocked Jurkat cells are examined. Four major hypersensitive sites at -120/-20 bp (HS1), +360 bp (HS2), +630/+780 bp (HS4) and around +1020 bp (HS5) with respect to the transcription start site of hsp90 beta gene have been identified. The HS1 is shared by both constitutive and heat shock, while the intronic sites of HS4 and HS5 are elicited by heat shock and HS2 is illustrated only in constitutive expression. In addition, distal HSs at around 8.7 kb upstream and 6.8 kb downstream are found in both constitutive and heat shock expression, which indicate that the boundaries of the hsp90 beta gene extend for some 16 kb. The HS patterns confirm chromatin regulation in the expression of hsp90 beta gene under various treatments. PMID- 12531244 TI - Activation of signal transducer and activator of transcription 3 in rat liver after heat shock and reperfusion stress. AB - Changes in transcription factors (TFs) accompany many types of cell stresses. By using electrophoretic mobility assays we show that the DNA binding of signal transducer and activator of transcription 3 (STAT3) is activated in rat liver by heat shock and ischemia-reperfusion. Northern blot and Western blot analysis reveal an increase of the mRNA and protein level of this transcription factor. Under both conditions the phosphorylation of pre-existing STAT3 is prompt and precedes the increase in the STAT3 protein. The activation: (1) is functional, i.e. is followed by the transcription of the target gene alpha(1)-acid glycoprotein (2) is strongly inhibited by pretreatment with the interleukin-1 receptor antagonist before heat shock but only slightly by pretreatment before ischemia-reperfusion (3) might, at least in part, be mediated by a cytokine cascade involving also interleukin-6. These results are consistent with the hypothesis that different kinds of stress can activate a number of common TFs. PMID- 12531245 TI - Retrovirus-mediated human heme oxygenase-1 (HO-1) gene transfer into rat endothelial cells: the effect of HO-1 inducers on the expression of cytokines. AB - The present study was conducted to investigate if the mechanism of human heme oxygenase-1 (HO-1) mediated angiogenesis was through the induction of vascular endothelial growth factor (VEGF). Also, the effect of HO-1 on the expression of transforming growth factor beta (TGF-beta),was studied in the presence and absence of HO-1 inducers. Rat lung microvessel endothelial cell line transduced with human HO-1 gene was subjected to cell culture (six separate experiments). mRNA extraction and reverse transcriptase polymerase chain reaction (RT-PCR) experiments, were performed to evaluate the expression of HO-1, VEGF, and TGF beta in the presence and absence of HO inducers including H(2)O(2), endotoxin and snake venom metalloproteinase with disintegrin like activity(SnMP). ELISA technique was performed to evaluate the levels of the studied growth factors. The results of the study showed over expression of VEGF in endothelial cells transduced with HO-1 compared to control non-transduced endothelial cells. On the other hand, the expression of TGF-beta and its protein level were markedly inhibited in HO-1 transduced endothelial cells compared to control non-transduced cells. Endotoxin and SnMP showed more prominent effect on the expression of VEGF and suppression of TGF-beta in HO-1 transduced endothelial cells, suggesting that their effect is most probably mediated through induction of HO-1. PMID- 12531246 TI - Lipopolysaccharide from Proteus mirabilis O29 induces changes in red blood cell membrane lipids and proteins. AB - Alterations in red blood cell (RBC) plasma membranes, i.e. in lipids and proteins, and osmotic fragility of these cells after treatment with Proteus mirabilis O29 endotoxin (lipolysaccharide (LPS)) were examined using a spin labelling method. At the highest concentration of LPS, insignificantly decreased fluidity of membrane lipids was observed. Changes in conformation of membrane proteins were determined by two covalently bound spin labels, 4-maleimido-2,2,6,6 tetramethylpiperidine-1-oxyl (MSL) and 4-iodoacetamido-2,2,6,6 tetramethylpiperidine-1-oxyl (ISL). The analysis of spectra of MSL and ISL showed modifications in membrane proteins in red blood cells treated with the highest concentration of lipopolysaccharide. On the other hand, in the case of isolated membranes, disturbances in membrane were observed for all concentrations of LPS. The alterations in membrane lipids and proteins are paralleled in a significant rise in osmotic fragility of RBCs upon endotoxin treatment. These results provide experimental evidence that P. mirabilis O29 LPS causes deleterious changes in membranes of human red blood cells. They show that action of lipopolysaccharide mainly concerns the membrane cytoskeleton. PMID- 12531247 TI - Decreased plasminogen activator inhibitor-1 secretion in hypoxic corneal epithelial cells is associated with increased urokinase plasminogen activator activity. AB - The aim of this study was to determine the effects of hypoxia on mRNA levels, cell-associated and -secreted protein concentration, activity, and protein complex formation of urokinase-type plasminogen activator, its receptor, and plasminogen activator inhibitor type-1 in corneal epithelium. Non-transformed human corneal epithelial cells were cultured in 20% oxygen (normoxic conditions) or 2% oxygen (hypoxic conditions) for 1, 3, 5, or 7 days. Relative changes in mRNA levels of plasminogen activator, receptor, and plasminogen activator inhibitor-1 were determined using a cDNA expression array, chemiluminescence, and densitometry. Protein concentrations were determined using enzyme linked immunosorbent assays. Activity assays were also used. Protein complex formation was assayed using cell surface biotinylation, immunoprecipitation, and Western blot analysis. Hypoxic corneal epithelial cells demonstrated no significant differences in plasminogen activator or receptor mRNA. Cell-associated plasminogen activator and membrane-associated receptor protein levels were unchanged. In contrast decreases in mRNA and secreted plasminogen activator inhibitor-1 protein were observed in hypoxic cells. Concurrently, increased cell associated plasminogen activator activity was observed in hypoxic cells. The formation of plasminogen activator/receptor/plasminogen activator inhibitor-1 complex at the cell surface was not inhibited by hypoxia. However, in hypoxic cells less plasminogen activator inhibitor-1 was associated with receptor. It is concluded that in corneal epithelium cultured in 2% oxygen plasminogen activator inhibitor-1 may be an important regulatory factor of the plasminogen activator system resulting in increased urokinase plasminogen activator activity. PMID- 12531248 TI - Oxidation of low-density lipoprotein by hemoglobin-hemichrome. AB - Hemoglobin and myoglobin are inducers of low-density lipoprotein oxidation in the presence of H(2)O(2). The reaction of these hemoproteins with H(2)O(2) result in a mixture of protein products known as hemichromes. The oxygen-binding hemoproteins function as peroxidases but as compared to classic heme-peroxidases have a much lower activity on small sized and a higher one on large sized substrates. A heme-globin covalent adduct, a component identified in myoglobin hemichrome, was reported to be the cause of myoglobin peroxidase activity on low density lipoprotein. In this study, we analyzed the function of hemoglobin hemichrome in low-density lipoprotein oxidation. Oxidation of lipids was analyzed by formation of conjugated diene and malondialdehyde; and oxidation of Apo-B protein was analyzed by development of bityrosine fluorescence and covalently cross-linked protein. Hemoglobin-hemichrome has indeed triggered oxidation of both lipids and protein, but unlike myoglobin, hemichrome has required the presence of H(2)O(2). In correlation to this, we found that unlike myoglobin, hemichrome formed by hemoglobin/H(2)O(2) does not contain a globin-heme covalent adduct. Nevertheless, hemoglobin-hemichrome remains oxidatively active towards LDL, indicating that other components of the oxidatively denatured hemoglobin should be considered responsible for its hazardous activity in vascular pathology. PMID- 12531249 TI - Melatonin preserves arachidonic and docosapentaenoic acids during ascorbate-Fe2+ peroxidation of rat testis microsomes and mitochondria. AB - The pineal hormone melatonin (N-acetyl, 5-methoxytryptamine) was recently accepted to act as an antioxidant under both in vivo and in vitro conditions. In this study, we examined the possible preventive effect of melatonin on ascorbate Fe(2+) lipid peroxidation of rat testis microsomes and mitochondria. Special attention was paid to the changes produced on the highly polyunsaturated fatty acids C20:4 n6 and C22:5 n6. The lipid peroxidation of testis microsomes or mitochondria produced a significant decrease of C20:4 n6 and C22:5 n6. The light emission (chemiluminescence) used as a marker of lipid peroxidation was similar in both kinds of organelles when the control and peroxidized groups were compared. Both long chain polyunsaturated fatty acids were protected when melatonin was incorporated either in microsomes or mitochondria. The melatonin concentration required to inhibit by 100% the lipid peroxidation process was 5.0 and 1.0mM in rat testis microsomes and mitochondria, respectively. IC 50 values calculated from the inhibition curve of melatonin on the chemiluminescence rates were higher in microsomes (4.98 mM) than in mitochondria (0.67 mM). The protective effect observed by melatonin in rat testis mitochondria was higher than that observed in microsomes which could be explained if we consider that the sum of C20:4 n6+C22:5 n6 in testis microsomes is two-fold greater than present in mitochondria. PMID- 12531250 TI - Acutely administered melatonin restores hepatic mitochondrial physiology in old mice. AB - Damage to mitochondria as a result of the intrinsic generation of free radicals is theoretically involved in the processes of cellular aging. Herein, we investigated whether acutely administered melatonin, due to its free radical scavenging activity, would influence mitochondrial metabolism. Mitochondrial respiratory activity and respiratory chain complex I and IV activities in liver mitochondria from a strain of senescence-accelerated-prone mice (SAMP8) and a strain of senescence-accelerated-resistant mice (SAMR1) were measured when the animals were 12 months of age. Respiratory control index (RCI), ADP/O ratio, State 3 respiration and dinitrophenol (DNP)-dependent uncoupled respiration were significantly lower in SAMP8 than in SAMR1. In contrast, State 4 respiration was significantly higher in SAMP8 than in SAMR1. Activities of complexes I and IV in SAMP8 were significantly lower than in SAMR1. Melatonin administration (10mg/kg body weight, intraperitoneally) 1h prior to sacrifice significantly increased RCI, ADP/O ratio, State 3 respiration and DNP-induced uncoupled respiration in SAMP8 while also significantly reducing State 4 respiration in SAMP8. The injection of melatonin also significantly increased complex I activity in both mouse strains and complex IV activity in the liver of SAMP8 mice. These results document an age-related decrease in hepatic mitochondrial function in SAM which can be modified by an acute pharmacological injection of melatonin; the indole stimulated mitochondrial respiratory chain activity which would likely reduce deteriorative oxidative changes in mitochondria that normally occur in advanced age. PMID- 12531251 TI - Altered content composition and structure of glycosaminoglycans and proteoglycans in gastric carcinoma. AB - Glycosaminoglycans (GAGs) in proteoglycan (PG) forms or as free GAGs are implicated in the growth and progression of malignant tumors. These macromolecules were investigated in human gastric carcinoma (HGC) and compared with those in human normal gastric mucosa (HNG). We report that HGC contained about 2-fold increased amounts of GAGs in comparison to HNG. Specifically, HGC showed 3- and 2.5-fold net increase in chondroitin sulphate (CS) and hyaluronan (HA) contents, respectively. Dermatan sulphate (DS) was slightly increased, but the amount of heparan sulphate (HS) was decreased. Of particular, interest were the quite different sulphation profiles of CS and DS chains in HGC in which, non sulphated and 6-sulphated disaccharide units were increased 10 and 4 times, respectively, in comparison to HNG. On PG level, three different populations were identified in both HNG and HGC, being HSPGs, versican (CS/DS chains) and decorin (CS/DS chains). In HGC, the amounts of versican and decorin were significantly increased about 3- and 8-fold, respectively. These PGs were also characterized by marked decrease in hydrodynamic size and GAG content per PG molecule. Analysis of Delta-disaccharide of versican and decorin from HGC showed an increase of 6 sulphated Delta-disaccharides (Delta di-6S) and non-sulphated Delta-disaccharides (Delta di-0S) with a parallel decrease of 4-sulphated Delta-disaccharides (Delta di-4S) as compared to HNG, which closely correlated with the increase of CS content. In addition, the accumulation of core proteins of versican and decorin in HGC was also associated with many post-translational modifications, referring to the number, size, degree and patterns of sulphation and epimerization of CS/DS chains. Studies on the modified metabolism of PGs/GAGs are under progress and will help in deeper understanding of the environment in which tumor cells proliferate and invade. PMID- 12531252 TI - Inhibitive effect of zinc ion on fatty acid synthase from chicken liver. AB - Fatty acid synthase (FAS; acyl-CoA:malonyl-CoA C-acyltransferase [decarboxylating, oxoacyl- and enoyl-reducing and thioester-hydrolyzing], EC 2.3.1.85) is an important enzyme participating in energy metabolism in vivo which is related to adiposis and cancer [Cancer Lett. 167 (1) (2001) 99; Nat. Med. 8 (4) (2002) 335]. Tests of fast- and slow-binding inhibitions showed that fatty acid synthase of chicken liver is rapidly and irreversibly inactivated by low Zn(2+) concentrations. Electrophoresis and FPLC results showed that FAS cross links occurred in the presence of high Zn(2+) concentrations (>4 microM) which may be another reason that FAS lost its activity. The modification velocity of FAS by DTNB decreased with increasing Zn(2+) concentration, which confirmed that Zn(2+) interacted with SH groups. Substrate protective experiments and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that all three substrates tested had some protective effects on FAS in the presence of Zn(2+), and malonyl-CoA was the most effective of the three substrates. In the presence of malonyl-CoA, the activity loss of FAS decreased sharply and almost no cross-link was observed in SDS-PAGE. This suggests that the phosphopantetheine SH group is the critical group in the cross-link and inhibition of FAS in the presence of Zn(2+). PMID- 12531253 TI - Nutrient cycling at the sediment-water interface and in sediments at Chiricahueto marsh: a subtropical ecosystem associated with agricultural land uses. AB - A study was conducted to examine the potential role to accumulate and transform nitrogen and phosphorus of a common wetland type marsh; the site is located in a subtropical environment that receives agricultural pollutants. Chiricahueto marsh effectively removes N and P from surface waters. It is clear that the diagenetic processes are mainly controlled by the exponential decomposition of organic matter that takes place at the sediment-water interface and in the sedimentary column, under oxic and suboxic-anoxic conditions, respectively. Mass balances indicated a net sedimentation of 11.3 g Nm(-2)x yr (-1) and 3.9 g Pm(-2) x yr ( 1), which results in an annual regeneration of 1.7 g Nm(-2) and 0.8 g Pm(-2) at the sediment-water interface under oxic conditions. A major remineralization rate was estimated in 6.4 g Nm(-2) x yr (-1) and 1.1g Pm(-2) x yr (-1) into the sedimentary column where suboxic to anoxic conditions occur by the utilization of nitrate, Fe and Mn as electron acceptors. The estimated burial fluxes in the deeper sediments (>50 cm) were 1.5 g Nm(-2) x yr (-1) and 0.4 g Pm(-2) x yr (-1). PMID- 12531254 TI - Uptake of heavy metals from synthetic aqueous solutions using modified PEI-silica gels. AB - The decontamination of synthetic Pb(II), Zn(II), Cd(II), Ni(II) solutions was investigated, using silica gels chemically modified with poly(ethyleneimine) (PEI) as sorbents. Two families of sorbents, i.e. silica/PEI and crosslinked silica/PEI, were prepared and characterized. Then the removal of metal ions from synthetic aqueous solutions was studied by static tests. They revealed that the sorption capacities depend on the pH, the initial concentration and to some extent on the nature of the metal. The recovery of the metal cations from the saturated sorbents was possible with diluted acid, only for the crosslinked supports. In this case, the regeneration and reuse without sorption decrease, was demonstrated. The presence of other cations (as Na(+), Ca(2+)) and metals does not affect the sorption capacities. PMID- 12531255 TI - Gas chromatographic-mass spectrometric method for the determination of bisphenol A and its chlorinated derivatives in urban wastewater. AB - The simultaneous determination of trace amounts of endocrine disruptors such as bisphenol A (BPA) and its monochloro, dichloro, trichloro and tetrachloro derivatives in wastewater has been developed using gas chromatography-mass spectrometry (GC-MS). Compounds were previously extracted from the aqueous samples using a liquid-liquid extraction procedure with a mixture of dichloromethane:carbon tetrachloride (25/75). After extraction, solvent was removed and a silylation step was carried out with N,O-bis(trimethylsilyl) trifluoroacetamide (BSTFA). The silylated compounds were identified and quantified by GC-MS using an HP1-MS column. The retention times were 6.64 min for BPA silylated, 7.26 min for Cl-BPA silylated, 7.99 min for Cl(2)-BPA silylated, 8.85 min for Cl(3)-BPA silylated and 9.95 min for Cl(4)-BPA silylated. A clean-up is not necessary using SIM mode. Deuterated anthracene (2H(10)-anthracene) was used as an internal standard. The detection limits obtained were 0.3, 0.6, 2.0, 4.5 and 13.0 ng L(-1) for silylated BPA, Cl-BPA, Cl(2)-BPA, Cl(3)-BPA and Cl(4) BPA, respectively. The proposed method was applied satisfactory to the determination of these chemicals, in different types of wastewater previously spiked with different amounts of these chemicals at concentration levels ranging from 0.01 to 2.50 microg L(-1) for BPA, 0.05-2.50 micro L(-1) for Cl-BPA and 0.05 5.00 microg L(-1) for Cl(2)-BPA, Cl(3)-BPA and Cl(4)-BPA, respectively. The method was validated following standard addition methodology. PMID- 12531256 TI - Effect of fulvic acid on the sorption of Cu and Pb onto gamma-Al2O3. AB - This work investigated the adsorption of Cu and Pb at the surface of gamma Al(2)O(3) in the presence of fulvic acid to address the significance of dissolved organic matters on metal partitioning. Fulvic acid, obtained from International Humic Substance Society, represented dissolved organic matter. Fulvic acid concentrations employed herein were 1, 5, and 10 mg C/L, which simulated the relevant environmental conditions. Ion selective electrodes were employed to ascertain free Cu and Pb measurements. The maximum adsorption of 10mg C/L fulvic acid on gamma-Al(2)O(3) was 5 x 10(-2)mgC/mg gamma-Al(2)O(3). Fulvic acid promoted Cu adsorption in low pH conditions. The effects of fulvic acid on Pb adsorption were similar to those of Cu. The conditional stability constants of sorbed fulvic acids with Cu and Pb were determined to be in the order of 4 to 6 (log K). Cu and Pb species were modeled in heterogeneous systems using triple layer model. Simulation results indicated that metal species are dominantly in complexation with fulvic acid, both in solution and at the gamma-Al(2)O(3) surface. PMID- 12531257 TI - Virus removal during simulated soil-aquifer treatment. AB - Removals of indigenous coliphage and seeded poliovirus type 1 during simulated soil-aquifer treatment were evaluated during transport of secondary effluent under unsaturated flow conditions in 1-m soil columns. Independent variables included soil type (river sand or sandy loam) and infiltration rate. Removal of coliphage was in all cases less than removal of poliovirus type 1 (strain LSc 2ab), supporting contentions that indigenous coliphage can act as a conservative indicator of groundwater contamination by viral pathogens of human origin. Coliphage retention was significantly more efficient (p<0.001) in the finer grained sandy loam (93%) than in sand (76%). Increasing reactor detention time from 5 to 20 h increased coliphage attenuation from 70% to 99% in a 1-m sand column. There was a significant linear correlation (p=0.012) between log transformed (fractional) coliphage concentration [log(C/C(0))] and reactor detention time. Re-mobilization of attached coliphage occurred during simulated rainfall using low-ionic-strength water. Inhibition of aerobic respiration resulted in significantly less efficient coliphage attenuation (p=0.033), suggesting the involvement of aerobic microorganisms in the survival/retention of this virus. PMID- 12531258 TI - Application of 'waste' metal hydroxide sludge for adsorption of azo reactive dyes. AB - The capacity and mechanism of metal hydroxide sludge in removing azo reactive dyes from aqueous solution was investigated with different parameters, such as charge amount of dyes, system pH, adsorbent particle size, and adsorbent dosage. The three anionic dyes used were CI Reactive Red 2, CI Reactive Red 120, and CI Reactive Red 141, increasing in number of sulfonic groups, respectively. Only 0.2% (w/v) of powdered sludge (<75microm) achieved color removal from 30 mg l(-1) reactive dye solutions within 5 min without pH adjustment. The larger the charge amount of the dyes, the greater the adsorption (>90%) on the metal hydroxide sludge. The system pH played a significant role in the adsorption on metal hydroxides and formation of dye-metal complexes. The optimum system pH for dye adsorption was 8-9 which was close to the pH(zpc) of the sludge while the precipitation of dye-metal complexes occurred at system pH 2. The maximum adsorption capacity (Q degrees ) of the sludge for the reactive dyes was 48-62 mg dye g(-1) adsorbent. The Langmuir and Freundlich models showed that the higher charged dyes had a higher affinity of adsorption. The smaller particle size and the greater amount of adsorbent showed the faster process, due to an increase in surface area of adsorbent. Desorption studies elucidated that metal hydroxide sludge had a tendency for ion exchange adsorption of sulfonated azo reactive dyes. Leaching data showed that the treated water was nontoxic at a system pH above 5 or a solution pH above 2. PMID- 12531259 TI - Elucidating competitive adsorption mechanisms of atrazine and NOM using model compounds. AB - Based on the relative adsorbability of natural organic matter (NOM) fractions with different molecular weights (MWs), two model compounds, poly(styrene sulfonate) (PSS) (nominal MW=1800 Dalton) and p-dichlorobenzene (DCB), were chosen to study the competitive effect of large and small NOM molecules on atrazine adsorption by two powdered activated carbons (PACs) with different pore size distributions. Both isotherm and kinetic tests of atrazine adsorption were conducted using fresh PAC and PAC preloaded with the model compounds. The model compounds were found to affect atrazine adsorption through two different mechanisms due to their size difference: direct competition for sites by p-DCB and pore constriction/blockage by PSS-1.8k. p-DCB was found to significantly reduce atrazine adsorption capacity but to have no effect on atrazine adsorption kinetics. In contrast, the effect of PSS-1.8k on atrazine adsorption capacity was very small. Furthermore, during simultaneous adsorption, PSS-1.8k had no effect on atrazine surface diffusion. However, preloading PAC with PSS-1.8k lowered the atrazine surface diffusion coefficient, D(s), by more than three orders of magnitude; D(s) decreased with increasing solid phase PSS-1.8k concentration. The pore size distribution of the PAC was found to play an important role in competitive adsorption. A high mesopore surface area could alleviate pore blockage significantly. PMID- 12531260 TI - Crossflow electro-microfiltration of oxide-CMP wastewater. AB - In this work, an electrically enhanced crossflow microfiltration (EECMF) system was used for the treatment of oxide-CMP wastewater under different operating conditions to investigate their effects on filtration rate. Oxide-CMP wastewater was obtained from a wafer fab and characterized by various standard methods. A membrane with a cut size of 0.1microm was used in the EECMF system. Operating parameters studied include crossflow velocity, transmembrane pressure, mode of electric field, electric field strength, and mode of backwashing. The filtration rate was found to increase with increasing crossflow velocity, electric field strength, and transmembrane pressure when operated below the critical electric field strength. Experimental results indicated that intra-filtration-run backwashing outperformed inter-filtration-run backwashing in terms of filtration rate. Experimental results also showed that the filtration rate for pulsed mode operation was greater than that of no electric field application, but worse than that of continuous mode operation. Using the present EECMF system, a fairly good quality of filtrate was obtained. It was found that the filtrate having a turbidity of 0.39 NTU is achievable. Thus, the filtrate could be reused for non process applications. PMID- 12531261 TI - Catalytic wet air oxidation of phenol by CeO2 catalyst--effect of reaction conditions. AB - The effect of catalyst loading, oxygen pressure, reaction temperature and phenol concentration on phenol conversion and total organic carbon (TOC) conversion, using CeO(2) as the catalyst, was investigated. There appeared a maximum rate of phenol conversion and TOC conversion as the catalyst loading increased. With phenol concentrations in the range of 400-2500 mg/L and oxygen pressure of 0.5 or 1.0 MPa, the optimal catalyst loading was 1.0 g/L, while it was 2.0 g/L at an oxygen pressure of 1.5 MPa. With a phenol concentration of 5000 mg/L, the optimal loading was 2.0 g/L for all oxygen pressures tested. Catalyst loading influences the reaction via the free-radical chain reaction involved in the catalytic wet air oxidation of phenol. Regarding oxygen pressures, at a phenol concentration of 400mg/L, the influence of the tested pressures (0.5, 1.0 and 1.5 MPa) on the 3h conversion of phenol was negligible, while the effect was significant for higher concentrations of phenol. The effect of oxygen pressure on TOC conversion was more profound, especially at a higher phenol concentration. At a pressure of 0.5 MPa, except for concentration of 400mg/L, the CO(2) selectivity barely exceed 80% at best, and was less than 25% with a phenol concentration of 5000 mg/L. At a pressure of 1.5 MPa, the selectivity was as high as 90% even for a concentration of 5000 mg/L. As was expected, increase of reaction temperature shortened the time taken to reach 50% phenol conversion. In addition, TOC conversion also increased with reaction temperature. Working from these observed results, optimal operating conditions were proposed. PMID- 12531262 TI - Decision support for the selection of an appropriate in-lake measure to influence the phosphorus retention in sediments. AB - Many in-lake measures which aimed to influence the phosphorus retention in lake sediments have failed to improve the trophic state of the lakes. The present paper introduces a systematic approach to select an appropriate in-lake measure. Before selecting an in-lake measure the goal of the measure should be defined, the problems of the lake must be identified, and the probability of success must be estimated. The proposed decision support consists of two parts. Part A, pre selection, excludes inappropriate measures. It uses six criteria, which are mainly based on a simple mass-balance model, and the targets of restoration. The criteria describe the magnitude of the external versus the internal phosphorus loading, the dynamics of the internal load, and the lake morphometry. Each measure is weighted differently with respect to importance and suitability by specific quantified limits. Part B, selection, uses qualitative criteria, which are specific to the measure in question. Checking these criteria will help to select a measure with a low risk of failure. The suggested decision support is illustrated in flow charts and exemplified by Lake Arendsee in Germany. PMID- 12531263 TI - Study of heavy metal pollution and speciation in Buyak Menderes and Gediz river sediments. AB - In this study, two economically important rivers of Turkey, Gediz and Buyuk Menderes (BM) are studied to determine their environmental pollution levels. An old analytical procedure involving sequential chemical extraction is improved and used for the partitioning of particulate trace metals (Cu, Co, Cr, Mn, Fe, Zn, Pb and Ni). Cationic and anionic Mn and Cr species with different phases are also determined by using leaching, extraction and ion exchange speciation processes. The sediment samples are analysed using graphic-furnace atomic absorption spectroscopy and differential pulse anodic stripping voltameter. Experimental results obtained on five replicate samples of fluvial bottom surface sediments at the sampling points demonstrate that the relative standard deviation of the sequential extraction procedure is generally better than +/-10%. The accuracy, determined by comparing total metal concentrations with the sum of the five sequential chemical extractions, is proved to be satisfactory. The detection limits established for three standard deviations of blank for different metals are identical and found to be 0.1mg/kg for sediment samples and 1 ppb for water samples. The results show that the pollution levels are significant especially for Pb, Cr, Mn and Zn in the Gediz river and Co, Mn, and Zn in the BM river. Comparison between our results and the measurements outlined before industrialization and the beginning of the intensive pesticide applications in agricultural fields (In: Broekaert et al., editors, Metal speciation in the environment, vol. 23. 1989. p. 601-11; A research on the environmental pollution in the agricultural fields and watering in the Aegean region, Ege University Research Project, No. 127 1988 (in Turkish); DSI Water Analyses Report, Ankara, 1985 (in Turkish)) show that the pollution in these rivers is probably originated from industrial, agricultural and domestic waste discharges. PMID- 12531264 TI - Simulating dynamic load of naturally occurring TOC from watershed into a river. AB - Naturally occurring total organic carbon (TOC) is an important feature of stream water quality. This study investigates the dynamic load of TOC from the deep creek watershed into the lower St. Johns River (LSJR), FL, USA, using numerical simulations and field measurements. An existing St. Johns River watershed assessment model for simultaneous loading of nutrients from watersheds into rivers is modified to include the TOC component for the purpose of this study. Three simulation scenarios (i.e., daily, monthly, and annually) are performed to estimate the dynamic load of TOC in response to rainfall events. Simulations show that rainfall events have decisive effects on TOC loads from the Deep Creek watershed into the LSJR. In general, the highly frequent the rainfall events occur, the higher the TOC loads into the river. Simulations also illustrate that effects of rainfall events on daily changes of TOC are minimal in winter, but are profound in late summer. Results suggest that TOC load into the river is not only a rainfall-driven but also a temperature-driven biological process. PMID- 12531265 TI - Inactivation of Bacillus subtilis spores with ozone and monochloramine. AB - The inactivation kinetics of Bacillus subtilis spores with ozone and monochloramine was characterized by a lag phase followed by a pseudo-first-order rate of inactivation. The lag phase decreased and the post-lag phase rate constant increased with increasing temperature within the range investigated (1 30 degrees C for ozone, 1-20 degrees C for monochloramine). The corresponding activation energies were 46820 J/mol for ozone and 79640 J/mol for monochloramine. The CT concept was found to be valid within the concentration range investigated of 0.44-4.8 mg/l for ozone, and 3.8-7.7 mg/l as Cl(2) for monochloramine. The inactivation kinetics of B. subtilis spores with both ozone and monochloramine varied with pH within the range of pH 6-10 investigated. The fastest ozone and monochloramine inactivation rates were observed at pH 10 and 6, respectively. Different stocks of the same strain of B. subtilis spores had different resistance to ozone and monochloramine mainly because of discrepancies in the extent of the lag phase. B. subtilis spores might not be conservative surrogates for C. parvum oocysts for ozone disinfection at relatively low temperature mainly due to the spores having a lower activation energy compared to that for the oocysts. In contrast, the activation energy for monochloramine was comparable for both microorganisms but differences in the extent of the lag phase might result in the spores being overly conservative surrogates for the oocysts at relatively low temperature. PMID- 12531266 TI - Use of complexing agents for effective ion-exchange separation of Co(II)/Ni(II) from aqueous solutions. AB - Cation-exchange separation of Co(2+)/Ni(2+) from aqueous solutions using water soluble complexing agents of ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), iminodiacetic acid (IDA), and citrate was experimentally studied at 298 K. Experiments were carried out as a function of initial aqueous pH (1.0-6.0), concentration of total metals (1.5-45.0 mol/m(3)), the concentration ratio of two metals (0.1-10) and of complexing agent to the total metals (0-1). It was shown that the exchange selectivity strongly depended on solution pH and was not completely related to the affinity of any metal with the complexing agents. When a certain level of complexing agent was present, highly effective separation could be achieved at an appropriate pH range (for an equimolar metal solution, e.g., pH 2-3 with EDTA and NTA as well as pH>3 with IDA and citrate). The application potential of this method was highlighted for the separation of Co(2+) from binary mixtures in the presence of trace amount of Ni(2+) due to its high selectivity and the smaller amount of the complexing agents needed. PMID- 12531267 TI - Factors influencing deterioration of denitrification by oxygen entering an anoxic reactor through the surface. AB - The purpose of the paper is to examine the factors that influence the deterioration of denitrification in open anoxic reactors. For this investigation an ASM 1-based simulation model was developed and successfully applied to fit data from batch experiments carried out in lab-scale reactor vessels (uncovered and covered) using both clarified domestic wastewater and synthetic wastewater. Applying the verified model, simulation studies were performed to investigate the effects of available denitrifiable substrate, biomass concentration, oxygen transfer rate, and temperature on deterioration of denitrification in open anoxic reactors. It has been shown that oxygen entering an anoxic reactor through the surface may not just affect denitrification metabolically, but also kinetically, due to increased dissolved oxygen (DO) concentration exerting an inhibitory effect on the denitrification rate. When the exogenous substrate concentration in the reactor vessel is high enough for a high consumption rate, the DO concentration is kept low. The higher the biomass concentration, and thereby the consumption rate of endogenous substrate, the lower the DO concentration during the low-rate denitrification phase. At low substrate removal rates, decreasing temperature will cause the DO concentration in anoxic vessels to increase. The results suggest that assuring removal of available exogenous carbon source at high rate by staging of open anoxic bioreactors may significantly improve denitrification efficiency. PMID- 12531268 TI - Selection of NF membrane to improve quality of chemically treated surface water. AB - The requirement for higher quality drinking water necessitates the application of more efficient water treatment techniques. Nanofiltration is one promising option for enhanced water treatment, for example, in enhanced organic matter removal. The characteristics of different nanofiltration membranes vary remarkably, and the selection of a membrane has to be made according to the requirements of an application. In this study six nanofiltration membranes (NF70, NF255, NTR-7450, NTR-7410, Desal-5 and TFC-S) were evaluated in improving the quality of chemically pre-treated surface water in a pilot-scale process. The results indicate that the membrane with high organics removal and slightly reduced ion removal characteristics (NF255) performed best in terms of product water quality as well as membrane productivity and fouling. The most permeable membrane (NTR 7410) suffered intensive fouling and insufficient product water quality. An interesting finding was that the permeates of all the tested membranes possessed a significant potential for microbial growth, despite the low nutrient contents. PMID- 12531269 TI - Changes in fractal dimension during aggregation. AB - Experiments were performed to evaluate temporal changes in the fractal dimension of aggregates formed during flocculation of an initially monodisperse suspension of latex microspheres. Particle size distributions and aggregate geometrical information at different mixing times were obtained using a non-intrusive optical sampling and digital image analysis technique, under variable conditions of mixing speed, coagulant (alum) dose and particle concentration. Pixel resolution required to determine aggregate size and geometric measures including the fractal dimension is discussed and a quantitative measure of accuracy is developed. The two-dimensional fractal dimension was found to range from 1.94 to 1.48, corresponding to aggregates that are either relatively compact or loosely structured, respectively. Changes in fractal dimension are explained using a conceptual model, which describes changes in fractal dimension associated with aggregate growth and changes in aggregate structure. For aggregation of an initially monodisperse suspension, the fractal dimension was found to decrease over time in the initial stages of floc formation. PMID- 12531270 TI - Chemistry of trace elements in coalbed methane product water. AB - Extraction of methane (natural gas) from coal deposits is facilitated by pumping of aquifer water. Coalbed methane (CBM) product water, produced from pumping ground water, is discharged into associated unlined holding ponds. The objective of this study was to examine the chemistry of trace elements in CBM product water at discharge points and in associated holding ponds across the Powder River Basin, Wyoming. Product water samples from discharge points and associated holding ponds were collected from the Cheyenne River (CHR), Belle Fourche River (BFR), and Little Powder River (LPR) watersheds during the summers of 1999 and 2000. Samples were analyzed for pH, Al (aluminum), As (arsenic), B (boron), Ba (barium), Cr (chromium), Cu (copper), F (fluoride), Fe (iron), Mn (manganese), Mo (molybdenum), Se (selenium), and Zn (zinc). Chemistry of trace element concentrations were modeled with the MINTEQA2 geochemical equilibrium model. Results of this study show that pH of product water for three watersheds increased in holding ponds. For example the pH of CBM product water increased from 7.21 to 8.26 for LPR watershed. Among three watersheds, the CBM product water exhibited relatively less change in trace element concentrations in CHR watershed holding ponds. Concentration of dissolved Al, Fe, As, Se, and F in product water increased in BFR watershed holding ponds. For example, concentration of dissolved Fe increased from 113 to 135 microg/L. Boron, Cu, and Zn concentrations of product water did not change in BFR watershed holding ponds. However, concentration of dissolved Ba, Mn, and Cr in product water decreased in BFR watershed holding ponds. For instance, Ba and Cr concentrations decreased from 445 to 386 microg/L and from 43.6 to 25.1 microg/L, respectively. In the LPR watershed, Al, Fe, As, Se, and F concentrations of product water increased substantially in holding ponds. For example, Fe concentration increased from 192 to 312 microg/L. However, concentration of dissolved Ba, Mn, Cr, and Zn decreased in holding ponds. Geochemical modeling calculations suggested that observed increase of Al and Fe concentrations in holding ponds was due to increase in concentration of Al(OH)(4)(-) and Fe(OH)(4)(-) species in water which were responsible for pH increases. Decreases in Ba, Mn, Cr, and Zn concentrations were attributed to the increase in pH, resulting in precipitates of BaSO(4) (barite), MnCO(3) (rhodochrosite), Cr(OH)(2) (chromium hydroxide), and ZnCO(3) (smithsonite) in pond waters, respectively. PMID- 12531271 TI - Mechanisms for naphthalene removal during electrolytic aeration. AB - Batch tests were performed to investigate chemical and physical processes that may result during electrolytic aeration of a contaminated aquifer using naphthalene as a model contaminant. Naphthalene degradation of 58-66% took place electrolytically and occurred at the same rates at a pH of 4 and 7. 1,4 naphthoquinone was identified as a product of the electrolysis. Stripping due to gases produced at the electrodes did not result in any naphthalene loss. Hydrogen peroxide (which may be produced at the cathode) did not have any effect on naphthalene, but the addition of ferrous iron (which may be present in aquifers) resulted in 67-99% disappearance of naphthalene. Chlorine (which may be produced from the anodic oxidation of chloride) can effectively degrade naphthalene at pH of 4, but not at a pH of 7. Mono-, di- and poly chloronaphthalenes were identified as oxidation products. Ferric iron coagulation (due to the oxidation of ferrous iron) did not significantly contribute to naphthalene loss. Overall, electrolytic oxidation and chemical oxidation due to the electrolytic by-products formed are significant abiotic processes that could occur and should be accounted for if bioremediation of PAH-contaminated sites via electrolytic aeration is considered. Possible undesirable products such as chlorinated compounds may be formed when significant amounts of chlorides are present. PMID- 12531272 TI - Anaerobic treatment of wastewater with high organic content using a stirred tank reactor coupled with a membrane filtration unit. AB - Using a cross-flow membrane bioreactor, high anaerobic conversion rates of three different types of wastewater with varying organic content were achieved. Loading rates obtained were as follows: 20 g CODL(-1) x d(-1) for artificial wastewater, approximately 8 g CODL(-1) x d(-1) from vegetable processing industry (sauerkraut brine) and 6-8 g CODL(-1) x d(-1) for wastewater from an animal slaughterhouse. At stable conditions, COD-removal rates in all three wastewaters were higher than 90%. Methane yields from the treatment of artificial wastewater, sauerkraut brine, and animal slaughterhouse wastewater were in the range of 0.17-0.30, 0.20 0.34, and 0.12-0.32 L(n) x g(-1) COD(-1) fed, respectively. The complete retention of biomass and suspended solids is a unique feature of this treatment process, which combines a high loading capacity and at the same time, high COD removal rates even for complex wastewater containing high concentrations of particulate matter. PMID- 12531273 TI - Application of a rapid method for identifying fecal pollution sources in a multi use estuary. AB - We demonstrate the application of a new PCR assay to detect and differentiate human and ruminant sources of fecal pollution in natural water samples. We tested samples collected from Tillamook Bay, Oregon, which has a long history of fecal pollution levels that exceed acceptable standards. The most likely sources are from dairy operations and ineffective sewage treatment. Using a suite of three PCR primer pairs specific for human or ruminant bacterial 16S ribosomal DNA markers, we detected at least one marker in 17 of 22 samples. In general, host specific fecal markers were detected in areas that are heavily impacted by anthropogenic activities. Nine out of 11 sites classified as either urban or near a sewage point source were positive for the human marker while only five of these same sites were positive for ruminant markers. Conversely, 12 out of 21 sites classified as rural or agricultural use were positive for ruminant markers, while only six of these sites were positive for human pollution. This suite of host specific genetic markers holds promise for identifying non-point source fecal pollution in coastal waters. PMID- 12531274 TI - Addition of trace metals increases denitrification rate in closed marine systems. AB - We investigated the effect of trace metals (Fe, Mn, Cu, Zn and Mo) on the denitrification unit at the Montreal Biodome. Two dosages of the five trace metals were tested on a denitrifying bacterial population which was extracted from the denitrification unit and cultured in 250 mL chemostats with artificial seawater. The low dosage showed a 20% increase in the denitrification rate whereas the high dosage had a more pronounced effect with a 250% increase. No increase in bacterial growth was observed, suggesting that the trace metals had an effect on the denitrification activity. When the trace metals were tested separately, only iron had a significant effect similar to the increase in the denitrification rate observed when the five trace metals were added. The combination of Fe and Mn caused a small but significant increase compared to the five trace metals. We then tested the effect of adding Fe, Mn and Cu to the denitrification unit at the Montreal Biodome. A high dosage of these trace metals showed a 250% increase in the denitrification rate, which went from 200 to 700 g NO(x)-N/d. Our results showed that the addition of trace metals is crucial for denitrification activities. PMID- 12531275 TI - Fate of physical, chemical, and microbial contaminants in domestic wastewater following treatment by small constructed wetlands. AB - In order to evaluate the efficacy of constructed wetlands for treatment of domestic wastewater for small communities located in rural areas, small-scale wetland mesocosms (400 L each) containing two treatment designs (a mixture of Typha, Scirpus, and Juncus species; control without vegetation) were planted into two depths (45 or 60 cm) with pea gravel. Each mesocosm received 19 L/day of primary-treated domestic sewage. Mesocosms were monitored (inflow and outflow samples) on a monthly basis over a 2-year period for pH, total suspended solids (TSS), 5-day biochemical oxygen demand (BOD(5)), total Kjeldahl nitrogen (TKN), dissolved oxygen (DO), and conductivity. Microbiological analyses included enumeration of fecal coliforms, enterococci, Salmonella, Shigella, Yersinia, and coliphage. Significant differences between influent and effluent water quality for the vegetated wetlands (p<0.05) were observed in TSS, BOD(5), and TKN. Increased DO and reduction in fecal coliform, enterococcus, Salmonella, Shigella, Yersinia, and coliphage populations also were observed in vegetated wetlands. Greatest microbial reductions were observed in the planted mesocosms compared to those lacking vegetation. Despite marked reduction of several contaminants, wetland-treated effluents did not consistently meet final discharge limits for receiving bodies of water. Removal efficiencies for bacteria and several chemical parameters were more apparent during the initial year compared to the second year of operation, suggesting concern for long-term efficiency and stability of such wetlands. PMID- 12531276 TI - Evolution over time of the agricultural pollution of waters in an area of Salamanca and Zamora (Spain). AB - A survey of the herbicides present in surface and groundwaters was conducted in 1999 in an area of the provinces of Salamanca and Zamora (Central-Western Spain) to assess the degree of pollution of the agricultural land and seasonal changes in the presence of herbicide residues. Ten sites were sampled and screened for 17 herbicides commonly used in the area; the compounds were ureas, triazines, amides, and others. A previously optimised method involving solid-phase extraction with polymeric cartridges, followed by HPLC with diode array detection, was used to monitor the herbicides. Of the 17 compounds examined, eight were found: chlorotoluron (41% of total detections), terbutryn (21%), atrazine (14%), linuron (7%), isoproturon and metolachlor (5.5% each), lenacil (4%) and metamitron (2%). Of the detections, 66% corresponded to river water samples (three sites). The herbicides found in groundwaters (seven sites) were: chlorotoluron, atrazine, terbutryn, linuron, and isoproturon, all of which are classified as probable or transient leachers in Europe. The temporal evolution of the herbicide content in river waters shows that the observed pollution is a function of time and is related to the application and use of the herbicides. By contrast, the pollution found in groundwaters was lower than that seen in surface waters, except that due to chlorotoluron, which, additionally, remained almost constant throughout the study period. PMID- 12531277 TI - Natural organic matter and DBP formation potential in Alaskan water supplies. AB - Disinfection by-products (DBP) are formed when natural organic matter (NOM) in water reacts with a disinfectant, usually chlorine. DBPs are a health risk element and regulated under the Safe Drinking Water Act. A study was conducted to evaluate the characteristics of NOM that contribute to DBPs in 17 different drinking water systems in Alaska. In order to determine the nature of the organic matter contributing to DBPs, DBP formation potential was compared with standard water quality parameters such as UV-254, color and dissolved organic carbon (DOC), as well as pyrolysis-gas chromatography/mass spectrometry (GC/MS). Results showed strong correlations between UV-254 and DBP formation potential for all waters studied. DOC, on the other hand, was less strongly correlated to DBP formation potential. Unlike previous studies, the total trihalomethane and haloacetic acid formation potentials were equal on a mass concentration basis for the waters studied. Pyrolysis-GC/MS indicated that NOM contributing to DBPs were primarily phenolic compounds. This finding was consistent with previous studies; however, unlike other studies, no correlation was found between aliphatic compounds in the raw waters and DBP formation potential. PMID- 12531278 TI - Removal of copper, nickel, cobalt and manganese from aqueous solution by kaolinite. AB - The removal of some heavy metals such as Mn(II), Co(II), Ni(II), and Cu(II) from aqueous solution is studied using a raw kaolinite. The sorption of these metals on kaolinite conformed to linear form of Langmuir adsorption equation. Langmuir C(m) constants for each metal were found as 0.446 mg/g (Mn), 0.919 mg/g (Co), 1.669 mg/g (Ni), 10787 mg/g (Cu) at 25 degrees C, respectively. Also, kinetic and thermodynamic parameters such as enthalpy (deltaH), free energy (deltaG) and entropy (deltaS) were calculated and these values show that adsorption of heavy metal on kaolinite was an endothermic process and the process of adsorption was favoured at high temperatures. PMID- 12531279 TI - Haloacetic acids and trihalomethanes in finished drinking waters from heterogeneous sources. AB - Trihalomethanes (THM) and haloacetic acids (HAA) are the most frequent chlorination by-products (CBP) in finished drinking waters. Traditionally, THM have been used as surrogates for CBP although the quantitative association between THM and other CBP is not well established. This problem is addressed in the present study from the analysis of THM and HAA in drinking water samples from four Spanish regions, representing areas with very different CBP composition, e.g. between 86 and 8.0 microg/l of THM and 50-3.0 microg/l of HAA. The resulting dataset exhibit a statistically significant correlation between total THM and HAA (Pearson's correlation coefficient, r(p)=0.815,p<0.0005). Furthermore, specific HAA are highly correlated with specific THM or their combinations. Accordingly, multivariate linear regression analysis of the concentrations observed show that the levels in total and specific HAA can be predicted from the THM content. These results are relevant for epidemiological studies on health effects from CBP exposure since they usually involve comparison of populations consuming waters of very distinct quality. PMID- 12531280 TI - Allogeneic and syngeneic class I MHC complexes drive the association of CD8 and TCR on 2C T cells. AB - In most cases, cytotoxic T cell activation is dependent on the interaction of the T cell receptor (TCR) and CD8 with MHC class I molecules. In the CD8(+) T cell system based on the mouse cytotoxic T cell clone 2C, recognition of the allogeneic MHC L(d) exhibits a less significant role for CD8 than recognition of the syngeneic MHC K(b). Here, we examined whether this difference is related to the relative abilities of the two pepMHC complexes to drive the association of CD8 and TCR on the T cell surface. We show that both the syngeneic and allogeneic pepMHC induced association of CD8 and TCR, as revealed by fluorescence resonance energy transfer (FRET). Thus, the orientation of the syngeneic and allogeneic ligands when bound to the same TCR both allow CD8 to be recruited to the TCR complex. The conserved diagonal orientation of TCRs on different pepMHC ligands may facilitate such associations. The FRET results are consistent with the known binding properties and the CD8 involvement of the two different TCR:pepMHC interactions. PMID- 12531281 TI - Accessory proteins and the assembly of human class I MHC molecules: a molecular and structural perspective. AB - The cell-surface presentation of antigenic peptides by class I major histocompatibility complex (MHC) molecules to CD8+ T-cell receptors is part of an immune surveillance mechanism aimed at detecting foreign antigens. This process is initiated in the endoplasmic reticulum (ER) with the folding and assembly of class I MHC molecules which are then transported to the cell surface via the secretory pathway. In recent years, several accessory proteins have been identified as key components of the class I maturation process in the ER. These proteins include the lectin chaperones calnexin (CNX) and calreticulin (CRT), the thiol-dependent oxidoreductase ERp57, the transporter associated with antigen processing (TAP), and the protein tapasin. This review presents the most recent advances made in characterizing the biochemical and structural properties of these proteins, and discusses how this knowledge advances our current understanding of the molecular events underlying the folding and assembly of human class I MHC molecules in the ER. PMID- 12531282 TI - Expression of the FcRn receptor (alpha and beta) gene homologues in the intestine of suckling brushtail possum (Trichosurus vulpecula) pouch young. AB - The neonatal IgG transporter FcRn consists of two chains, FcRn alpha and beta (also known as beta(2) microglobulin), and is involved in transferring IgG molecules across both mammary and intestinal epithelial cells. Developmental changes in FcRn IgG alpha and beta chain mRNA levels were investigated in the gut of brushtail possum (Trichosurus vulpecula) pouch young (PY) using Northern hybridisation. FcRn alpha transcripts were detected in the PY proximal intestine at all times examined, between days 1 and 195 of post-natal life, with increased levels detected from around day 110. The beta(2) microglobulin transcript levels in the PY proximal intestine were low to undetectable until day 110 of post-natal life and then increased dramatically after day 159. Both the FcRn alpha and beta gene transcripts were detected in a wide range of tissues in the adult possum (>365 days). Genomic sequences located 5' to the start of transcription of the FcRn alpha and beta(2) microglobulin genes were cloned and analysed for predicted cis-acting transcription control elements. Both the FcRn alpha and beta(2) microglobulin genomic sequences contained STAT5 binding motifs consistent with the transcription of both genes being modulated by prolactin. Using in situ hybridisation, the FcRn alpha and beta(2) microglobulin transcripts were localised to the epithelial cells of the PY intestine. However, no prolactin receptor transcripts were detected in the same epithelial cells suggesting that the observed changes in FcRn alpha and beta(2) microglobulin gene expression in the proximal intestine are not modulated directly by prolactin. The results are consistent with the hypothesis that changes in FcRn alpha and beta(2) microglobulin gene expression take place in the possum PY intestine to accommodate changes in maternal milk composition to meet the changing immunological demands of the PY. PMID- 12531283 TI - Expression and characterization of recombinant canine IL-13 receptor alpha2 protein and its biological activity in vitro. AB - Our previous study showed that recombinant canine IL-13 (rcaIL-13) stimulated production of allergen-specific IgE in vitro by peripheral blood mononuclear cells (PBMC) from flea allergen-sensitized dogs. This has also been demonstrated using human IL-13 (huIL-13) and PBMC isolated from human allergy patients. The stimulatory activity of rcaIL-13 was specifically inhibited by a fusion protein of the extracellular domain of canine IL-13Ralpha2 and the Fc fragment of canine IgG heavy chain (rcaIL-13Ralpha2-Fc). In this communication, we report the construction and expression of a non-fused recombinant extracellular domain of canine IL-13Ralpha2 (rcaIL-13Ralpha2) in an E. coli expression system. The E. coli expressed rcaIL-13Ralpha2 was isolated in inclusion bodies, then solubilized in buffer containing denaturants and reducing agents. After refolding and purification, the biological activity of rcaIL-13Ralpha2 was found in the monomer fraction resulting from gel filtration and ion exchange chromatographies. Biological activity of purified rcaIL-13Ralpha2 was demonstrated by the specific inhibition of rcaIL-13 activity in a TF-1 cell proliferation assay. Additionally, rcaIL-13Ralpha2 was found to be active in neutralizing rcaIL-13 induced upregulation of IgE mRNA levels in PBMCs of "high IgE" dogs, which have been bred to exhibit a predisposition for high IgE production and are used as a model for allergic asthma. The data confirm our previous report that the regulatory effects of IL-13 on IgE production in canine PBMCs are similar to those reported in humans. Thus, allergic dogs, such as the "high IgE" producing dogs, may be excellent models for research on IgE-mediated diseases in humans. PMID- 12531284 TI - Comparison of three microbial hosts for the expression of an active catalytic scFv. AB - Antibodies represent an interesting protein framework on which catalytic functions can be grafted. In previous studies, we have reported the characterization of the catalytic antibody 4B2 obtained on the basis of the "bait and switch" strategy which catalyzes two different chemical reactions: the allylic isomerization of beta,gamma-unsaturated ketones and the Kemp elimination. We have cloned the antibody 4B2 and expressed it as a single-chain Fv (scFv) fragment in different expression systems, Escherichia coli and two yeasts species, in order to elicit the most suitable system to study its catalytic activity. The scFv4B2 was secreted as an active form in the culture medium of Pichia pastoris and Kluyveromyces lactis, which led respectively to 4 and 1.3mg/l after purification. In E. coli, different strategies were investigated to increase the cytoplasmic soluble fraction, which resulted, in all cases, in the expression of a low amount of functional antibodies. By contrast, substantial amount of scFv4B2 could be purified when it was expressed as inclusion bodies (12mg/l) and submitted to an in vitro refolding process. Its catalytic activity was measured and proved to be comparable to that of the whole IgG. However, the instability of the scFv4B2 in solution prevented from an exhaustive characterization of its activity and stabilization of this protein appears to be essential before designing strategies to improve its catalytic activity. PMID- 12531285 TI - Physical association and functional interaction between beta1 integrin and CD98 on human T lymphocytes. AB - CD98 is a cell surface protein previously characterized as a cell activation marker, an amino acid transporter, and has recently been implicated in integrin related functions. Integrins are cell surface proteins, important for homotypic cell aggregation, cell adhesion, and coactivation of T lymphocytes. We have previously shown that the anti-CD98 mAb 80A10, when coimmobilized with anti-CD3 mAb OKT3, is able to mediate human T cell coactivation that is inhibited by anti beta1 integrin specific mAb 18D3. These results indicated a functional association of CD98 and beta1 integrin signaling but left open the question of a physical association. We now show the induction of homotypic aggregation through CD98 among human T cells and this aggregation was inhibited by anti-beta1 integrin mAb. Therefore, CD98-dependent lymphocyte proliferation and adhesion may involve integrins. Competitive binding assays and fluorescence colocalization analysis suggested that CD98 and beta1 integrin were physically associated. Differential extraction techniques and immunoprecipitations provided the first evidence that the alpha4beta1 integrin and CD98 are specifically associated on human T lymphocytes. PMID- 12531286 TI - Elucidation of a downstream boundary of the 3' IgH regulatory region. AB - Class switch recombination (CSR) changes the immunoglobulin heavy chain (IgH) constant region gene (C(H)) in B cells from IgM to IgG, IgA, or IgE, without modifying the variable region gene segment. This process requires transcription through switch (S) regions located upstream of the C(H) genes targeted for CSR, a process that relies on the activity of an uncharacterized regulatory region at the 3' end of the C(H) locus (3' IgH RR) that has been implicated via the effects of pgk-neo cassettes inserted into the locus. The 30kb region just downstream of the most 3' C(H) gene (Ca) contains four known enhancer elements including HS3a, HS1,2, HS3b, and HS4. Replacement of either of the proximal two enhancer elements (HS3a or HS1,2) with a pgk-neo gene cassette disrupted germline transcription of and CSR to most C(H) genes. However, replacement of either of the enhancers with a loxP sequence had no effect on CSR indicating that these elements are not critical for CSR. Insertion of a pgk-neo cassette at various sites within the C(H) locus inhibited CSR to upstream, but not downstream C(H) genes, supporting the notion that the pgk-neo cassette insertion into the locus short-circuits the ability of the 3' RR to facilitate CSR of dependent C(H) genes upstream of the insertion. These analyses also indicated that the key elements of the 3' IgH RR were downstream from HS1,2. In this study, we have sought to localize the 3' IgH RR by defining its 3' boundary. For this purpose, a pgk-neo gene cassette was targeted 2kb downstream of the HS4 element in ES cells that had normal ability to undergo CSR. We then employed Rag-2 deficient blastocyst complementation to generate chimeric mice that harbored B cells homozygous for this mutation. Such chimeras exhibited normal reconstitution of the splenic compartment and had normal serum immunoglobulin levels. Upon in vitro activation, transcription from the pgk-neo cassette was induced in B cells, however, CSR to all measured IgH isotypes occurred at normal levels. These findings, coupled with previous pgk-neo insertion studies, suggest that key elements of the 3' IgH RR lie within a 17kb region between HS1,2 and 2kb downstream of HS4. PMID- 12531287 TI - Characterization of Cryptosporidium spp.--recent developments and future needs. AB - Cryptosporidia, widely distributed protozoan parasites of vertebrates have recently attracted increasing interest due to several serious waterborne outbreaks, the life-threatening nature of infection in immunocompromised patients, and the realization of economic losses caused by these pathogens in livestock. Genetic polymorphism within Cryptosporidium spp. is being detected at a continuously growing rate, owing to the widespread use of modern molecular techniques. The aim of this paper is to review the current status of taxonomy, genotyping, molecular phylogeny, and characterization of cryptosporidia, and to highlight the need for polyphasic typing, i.e. an integrated approach comprising standardized morphologic, biologic, and molecular methods for describing Cryptosporidium species and isolates, and for establishing "virtual" reference strains. PMID- 12531288 TI - The effect of Trypanosoma evansi infection on pig performance and vaccination against classical swine fever. AB - Although Trypanosoma evansi is not considered as an important pathogen in pigs, it may interfere with other pathogens or vaccinations by its immunosuppressive nature. In order to determine whether T. evansi alters pig performance and induces immunosuppression in pigs, induction of immune responses by vaccination against classical swine fever (CSF) and by immunization with a control antigen, human serum albumin (HSA), was assessed in T. evansi-infected and non-infected animals. Although T. evansi infection did not have a significant influence on growth performance, feed conversion or PCV, antibody responses against both the test antigen HSA and the CSF vaccine were significantly reduced in T. evansi infected animals as compared to uninfected animals. Moreover, the reduced response against the CSF vaccine appears to be accompanied by a less well developed protection against CSF with higher fever responses and leukopenia. This immunosuppression might explain the accounts of poor protection of CSF-vaccinated pigs reported in T. evansi-endemic areas of Vietnam, and suggests that prior treatments with trypanocidal drugs to improve the efficacy of CSF vaccination, may be justified. PMID- 12531289 TI - Responses of bovine chimaeras combining trypanosomosis resistant and susceptible genotypes to experimental infection with Trypanosoma congolense. AB - West African N'Dama cattle have developed a genetic capacity to survive, reproduce and remain productive under trypanosomosis risk. The cellular and molecular bases of this so-called trypanotolerance are not known, but the trait is manifested by the N'Dama's greater capacity to control parasitaemia and anaemia development during an infection. In order to examine the role of the haematopoietic system in trypanotolerance, we have exploited the tendency for the placentas of bovine twin embryos to fuse. Placental fusion in cattle results in bone marrow chimaerism in twins. By comparison with the N'Dama, cattle of the East African Boran breed are relatively susceptible. We evaluated the role of the haemopoietic system in trypanotolerance by comparing the performance of five Chimaeric Boran/N'Dama twin calves with that of singletons of the two breeds. Chimaeric Boran/N'Dama pairs of twins were produced in recipient Boran cows by embryo transfer, and the majority of haemopoietic cells in all twinned individuals were of Boran origin. Thus, N'Dama chimaeras differed from N'Dama singletons in that the bulk of their haemopoietic system was derived from their susceptible Boran twins, while Boran chimaeras differed little from Boran control animals. All cattle became parasitaemic and developed anaemia. The N'Dama chimaeras did not manage their anaemia and white blood cell counts effectively. However, they were able to limit parasitaemia development. These results suggest that trypanotolerance is the result of two mechanisms, one that improves parasite control and is independent of the genetic origin of the haemopoietic tissue, and another that is influenced by haemopoietic tissue genotype and which improves control over anaemia. The capacity to maintain growth during infection was similarly dependent on the genetic origin of the haemopoietic tissue. PMID- 12531290 TI - Evaluation by different diagnostic techniques of bovine abortion associated with Neospora caninum in Spain. AB - Eighty foetuses from some of the main cattle-producing regions in Spain were analysed to investigate the participation of Neospora caninum in cases of bovine abortion. Diagnosis of the infection was determined by histopathological analysis complemented with immunohistochemistry, serology (IFAT and ELISA) and PCR tests. A total of 38.8% of the bovine foetuses analysed were considered to be infected by at least one of the diagnostic techniques used. Microscopic lesions consistent with Neospora infection in brain were identified in 31.3% of the samples, whereas only 10.7 and 15.3% were positive using serological and PCR analysis, respectively. Perfect agreement was shown between IFAT and ELISA, although there was little agreement among results of the other diagnostic techniques. Gestational age of aborted foetuses checked ranged from <3 to 9 months, with a mean of 5.9 months, and no difference in age was found between infected and non infected foetuses (P>0.05). This study confirms the importance of N. caninum as a cause of abortion in Spain and underlines the need to use different diagnostic techniques to increase the chance to detect the infection in aborted foetuses. PMID- 12531291 TI - Utility of the Cryptosporidium oocyst wall protein (COWP) gene in a nested PCR approach for detection infection in cattle. AB - A preliminary molecular epidemiological study was carried out to investigate the utility of the Cryptosporidium oocyst wall protein (COWP) gene in the detection of Cryptosporidium oocysts in fecal samples. A nested polymerase chain reaction (PCR) approach using COWP gene primers was adopted for this purpose. Fecal samples were spiked with each of 1, 10, and 100 oocysts of C. parvum, four samples for each number, and the DNA was extracted from each sample using a glassbead method. The presence of oocysts was determined using the nested PCR with COWP gene primers, and the limit of detection of oocysts by the PCR was determined. The limit of detection was 100 oocysts spiked in 1 ml of fecal material (50% sold material) (four positives/four samples tested). Seventy-five percent of DNA extracted samples spiked with 1 and 10 oocysts was positive by the PCR (three positives/four samples tested). Based on this, small sample size using the COWP gene primers with a nested PCR analysis could reliably identify infected animals rather conveniently and accurately. PMID- 12531292 TI - Epidemiology of canine visceral leishmaniosis in the endemic area of Montes Claros Municipality, Minas Gerais State, Brazil. AB - The Montes Claros City is located in an endemic area for visceral leishmaniosis in the Minas Gerais State, Brazil. With the implementation of a program for the control of visceral leishmaniosis in 1994, a sectional study was carried out to evaluate the infection by viscerotropic Leishmania in the population of dogs from Montes Claros, basically using indirect immunofluorescence antibody test (IFAT). Blood samples were collected on filter paper from 33,937 dogs, representing 96.1% of the canine local population. The prevalence for visceral leishmaniosis was found to be 9.7% in the municipality, being 9.9% in the urban area and 8.8% in the rural area. The annual incidence showed to be 64.3/1000 dogs. Prevalence of infection was not correlated with dogs age. The most affected breeds were: Boxer (24.6%) and Cocker (26.9%); Mongrel dogs had a prevalence of 7.8%. Short-hair animals had a prevalence of 11.9%, while long-furred animals had a prevalence of 8.9%. The isoenzymatic profile indicated that Leishmania (Leishmania) chagasi was the visceral leishmaniosis etiological agent in Montes Claros City, Minas Gerais State, Brazil. The main geographical areas for the parasite transmission were identified, and control measures were immediately started. The role of the dog as a reservoir for L. chagasi was confirmed. It was demonstrated that short-furred animals are at a higher risk of acquiring visceral leishmaniosis than the long furred dogs. PMID- 12531293 TI - Echinococcus granulosus in Finland. AB - Echinococcus granulosus is shown to occur in eastern Finland in a sylvatic cycle involving wolves (Canis lupus) as the definitive host and reindeer (Rangifer tarandus tarandus) and probably also elk (Alces alces) and the wild forest reindeer (Rangifer tarandus fennicus) as intermediate hosts. Even though the prevalence of E. granulosus in reindeer has increased in recent years, it is still very low (<0.013%). The results suggest, however, that the reindeer is a good sentinel animal to show the contamination of soil with Echinococcus eggs. This is the first report of wolves acting as a definitive host for E. granulosus in northern Europe. The parasite seems to be quite common in the Finnish wolf population, with a prevalence of approximately 30% in both intestinal and faecal samples. The present results and previous ones indicate that E. granulosus infection has not spread to dogs in the reindeer herding area. PMID- 12531294 TI - Evaluation of gastro-intestinal nematode parasite control strategies for first season grazing cattle in Sweden. AB - A three-year grazing experiment (1998-2000) was conducted with first-season grazing cattle (FSGC) on improved pastures in central-eastern Sweden. Comparison was made between five groups with 10 calves in each group where four of these were set stocked and either (1) untreated, (2) ivermectin bolus treated, (3) subjected to biological control with the nematophagous fungus Duddingtonia flagrans, or (4) treated with a copper wire particle bolus. The fifth treatment was an evasive grazing strategy, whereby untreated calves were turned out onto pasture used by older cattle the previous year and then these calves were moved to silage aftermath in mid-July. To introduce low-levels of parasite infection to the experiment, each animal received a 'priming dose' of approximately 5,000 Ostertagia ostertagi and 5,000 Cooperia oncophora infective third stage larvae immediately prior to the start of the first grazing year of the trial. Results showed that efficient and sustainable parasite control of FSGC was possible to achieve without the use of anthelmintics by using turnout pastures that the previous year had been grazed by older cattle, in combination with a mid-July move to aftermath leys. Biological control also proved beneficial but the efficacy was impaired if high faecal egg counts coincided with rapid dung pat degradation due to heavy rainfall. No indication of parasite control was observed with the copper wire particle bolus. It was also demonstrated that the impact of gastrointestinal (GI) parasitism varied between years and that the level of overwintering contamination is important but likewise, is unpredictable. Although faecal egg counts in 1999 were low, due both to a delayed turnout and drought for the major part of the grazing season, deposited eggs successfully developed to infective larvae and overwintered in large numbers. The population of overwintered infective larvae at the time of turnout in early May played an important role in the course of infection in 2000 and resulted in an average 65 kg advantage of the ivermectin treated calves compared with the untreated calves.Thus, this three-year grazing experiment has emphasised the importance of subclinical gastrointestinal nematode infections in FSGC in Sweden. In addition, the study has shown that adequate parasite control may be achievable without the use of anthelmintics. PMID- 12531295 TI - Effect of moxidectin against natural infestation of the cattle tick Boophilus microplus (Acarina: Ixodidae) in the Mexican tropics. AB - The study was divided in to two trials and carried out in a ranch in eastern Yucatan state, Mexico. In the first trial, two groups of 15 BostaurusxBosindicus heifers, 6-12 month of age and naturally infested with Boophilus microplus ticks were used. Heifers in Group 1 were treated with a 1% injectable formulation of moxidectin at the dose of 0.20mg/kg body weight by subcutaneous injection. The other group remained as untreated controls. Number of immature and engorging female ticks were assessed on days 0, 7, 14, 21, 28 and 35 post-treatment (PT). The efficacy of moxidectin on adult ticks from day 7 to 28 PT was greater than 95%. The efficacy decreased to 74.9% by day 35. In the second trial, animals in Group 1 were treated with the moxidectin product as before, while cattle in Group 2 were treated according to the routine procedure for the control of ticks on that property (125 g/l amitraz as a dip). Treatment of all cattle was repeated four times at intervals of 28 days. The efficacy of the experimental moxidectin treatment was similar to that of the routine amitraz treatment, i.e., greater than 99%. PMID- 12531296 TI - Temporal pattern of isotype-specific antibody responses in primary and challenge infestations of sheep with Psoroptes ovis--the sheep scab mite. AB - In sheep Psoroptes ovis provokes an allergic dermatitis with significant P. ovis antigen-specific IgE responses. The kinetics of the IgE response to primary and challenge infestations of P. ovis were reported earlier [Parasite Immunol. 22 (2000) 407]. The present study examines IgG, IgM and IgA responses to primary and challenge infestations of P. ovis and the profile of antigens/allergens reacting with IgG and IgE antibodies. Antigen-specific enzyme-linked immunosorbent assays (ELISAs) demonstrate that primary infestations elicited significant increases in levels of IgG and IgM but not IgA antibodies. IgG and IgM responses to primary and challenge infestations were not significantly different. Western blots of reduced P. ovis proteins indicate that IgG antibodies reacted with five major antigens following primary infestation and only three of these after challenge infestation. IgE antibodies bound to three major and five minor allergens after primary infestation and two additional minor allergens after challenge infestation. Immunodominant antigens >100 and <15 kDa and allergens >100 kDa were most consistent in stimulating substantial IgG and IgE antibody responses, respectively. These antigens/allergens may be exploited in immunodiagnosis and modulation of the host immune response. PMID- 12531297 TI - Survey of ixodid tick species on domestic cats in Japan. AB - Various species of ixodid ticks, attached to domestic cats in Japan, were identified in spring (April to June) and autumn (September to November). In the spring, a total of 282 ticks, including 61 larvae, 70 nymphs, 127 females and 24 males were collected from 126 cats. Of these, 264 were identified up to the species level. In the spring, Haemaphysalis longicornis was the most frequently (39.7%, 50/126) found tick species on feline hosts, followed by Ixodes ovatus (35.0%, 44/126), Ixodes nipponensis (15.9%, 20/126) and Haemaphysalis flava (9.5%, 12/126). Small numbers of Haemaphysalis megaspinosa, Haemaphysalis japonica, Ixodes persulcatus, Ixodes granulatus and Amblyomma testudinarium were also recovered. H. longicornis was the most frequently found tick species on cats around riversides or river basins, while I. ovatus and I. nipponensis were more frequently found on cats kept near woodland or related areas. I. nipponensis was more frequently found on castrated males. No major statistical differences in the frequency of tick attachment among sex, age or hair length for the three major tick species were found. Of 205 ticks including 173 (84.4%) larvae, 27 (13.2%) nymphs, 4 (2.0%) females and 1 (0.5%) male recovered from 62 cats in autumn, only 32 (15.6%) were identified. Most of the larvae were fully- or partly-engorged Haemaphysalis spp., and it was difficult to identify them further by morphological characterization. PMID- 12531298 TI - Efficacy of formic acid in gel for Varroa control in Apis mellifera L.: importance of the dispenser position inside the hive. AB - The efficacy of formic acid in a gel matrix was evaluated in two groups of honeybee colonies. In Group 1, a dispenser with 120 g of formic acid (70%) in gel was placed on the brood combs and another dispenser with the same dose was located on the hive bottom (total dose, 240 g). Group 2 received two doses of 240 g of formic acid (70%) in gel and each application was applied in two dispensers containing 120 g of the formic acid solution each and they were located over the brood chamber (total dose, 480 g). In Group 2, the period between both applications was 15 days, and the efficacies after the first and both applications were calculated. Significant differences were registered for final efficacy between both groups. When final efficacy of Group 1 was compared with efficacy after first application of Group 2, significant differences were found (P=0.0005). Same doses in different positions within the hive have different final efficacy. The higher efficacy was registered when the dispensers were placed over brood combs and on the hive bottom. It is suggested that efficacy is related to dispenser position within the hive. PMID- 12531299 TI - Antibodies to Anaplasma marginale major surface proteins 1a and 1b inhibit infectivity for cultured tick cells. AB - Major surface protein 1 (MSP1) of the cattle pathogen Anaplasma marginale (Rickettsiales: Anaplasmataceae) is a complex of two proteins, MSP1a and MSP1b. Previous studies demonstrated that MSP1a and MSP1b are adhesins for bovine erythrocytes, while only MSP1a proved to be an adhesin for tick cells. In this study, a tick cell culture system for propagation of A. marginale was used to develop an infection inhibition assay for testing the ability of antisera to block infection of A. marginale for cultured tick cells. A. marginale derived from cell culture was incubated with various antisera prior to inoculation onto cell monolayers. The monolayers were harvested 7 days post-inoculation and A. marginale in the cultures was quantified using an antigen detection ELISA. Antisera tested in the infection inhibition assay were derived from persistently infected cattle, from cattle immunized with A. marginale purified from bovine erythrocytes, and from rabbits and cattle that were immunized with the recombinant MSP1a, MSP1b and MSP1 complex. Antibodies from cattle persistently infected with A. marginale, cattle immunized with A. marginale from bovine erythrocytes or cattle immunized with the recombinant MSP1 complex did not inhibit the infectivity of A. marginale for tick cells. Antiserum from rabbits immunized with MSP1a and MSP1b (individually or combined) reduced infection of both the Virginia and Oklahoma isolates of A. marginale for tick cells by 25-70%. Likewise, antisera from cattle immunized with recombinant MSP1a or MSP1b inhibited infection of tick cells by 26-37%. These results further confirm the role of MSP1 complex proteins in infection of tick cells. Lack of inhibition of infection by antisera from naturally infected cattle or cattle immunized with whole organisms suggests that the bovine immune response is not directed toward blocking infection of A. marginale for tick cells and may contribute to the continued infectivity of the pathogen for ticks. PMID- 12531300 TI - Relationship between liver disorders and protection against Eimeria stiedai infection in rabbits immunized with soluble antigens from the bile of infected rabbits. AB - Soluble antigens exist in the bile of rabbits infected with Eimeria stiedai (E. stiedai) in the acute phase, and rabbits immunized with the antigens show resistance against the infection. In this study, the liver function of rabbits immunized either with the soluble antigens or PBS were examined following the parasite challenge. Rabbits immunized with PBS shed a number of oocysts and showed an increase in r-glutamyltransferase (GGT) activity and a decrease in blood Indocyanine green (ICG) clearance. However, rabbits immunized with the soluble antigens shed a lower number of oocysts and showed a transient increase of alanine-aminotransferase (ALT) activity on Day 8 post-challenge (p.c.). The blood Indocyanine green clearance of the rabbits showed no change throughout the experiment. By histopathological observation of the liver, a number of merozoites were found in the biliary ducts on Day 8 post-challenge in the non-immunized rabbits. In contrast, a number of lymphocytes and neutrophilic leukocytes assembled around the biliary ducts of the immunized rabbits, but few parasites were found there on Day 8 post-challenge. These results suggest that the soluble antigens stimulate local immune reactions, for example around the biliary ducts, resulting in elimination of the parasite's development. PMID- 12531301 TI - Diurnal variation in microfilaremia in a cat experimentally infected with larvae of Dirofilaria immitis. AB - In five healthy mongrel female cats used, four cats (Cats 1-4) were experimentally inoculated with 100-123 larvae (L(3)) of Dirofilaria immitis. Cat 5 was an uninfected control. Only Cat 1 became microfilaremic on Day 201 after inoculation and the diurnal changes in the microfilaria population were monitored every 2 h for 24 h on Day 237 when a sufficient number of microfilaria were detected in the circulation. The maximum number of microfilaria in the blood (1,350/ml) occurred at 9:00 p.m. and then gradually decreased to the minimum of 300/ml at 7:00 a.m., indicating that the number of microfilaria shows a nocturnal sub-periodic pattern of diurnal rhythm even in peripheral blood of cats as an abnormal host. On postmortem examination, 10 live adult worms (three males and seven females) in Cat 1 and one live adult worm (one male) in Cat 2 were detected. PMID- 12531303 TI - Surface metal adsorption on zooplankton carapaces: implications for exposure and effects in consumer organisms. AB - The current study aimed to determine the potential of two important aquatic invertebrate crustacean species, Daphnia magna and Ceriodaphnia dubia, to adsorb cadmium on to their carapaces from aqueous solution. Using the Langmuir equation to model data outputs, it was shown that cadmium readily became associated with the carapace surfaces of both species, with uptake being dependent on exposure time and concentration. Maximum carapace-adsorption potential was found to be directly related to surface area, so that at predicted carapace saturation, D. magna neonates bound approximately five times more cadmium than the smaller C. dubia neonates. However, adsorption per unit surface area was found to be similar under the same exposure conditions. Results of surface metal adsorption studies in C. dubia suggested that short term exposures to high concentrations of aqueous cadmium would lead to similar levels of adsorption as obtained with long-term exposures to low concentrations. The study illustrates that contaminants adsorbed to prey surfaces may be an important mechanism of exposure to predators, and highlights some potential problems of feeding organisms during long-term toxicity tests. PMID- 12531304 TI - An automated overlying water-renewal system for sediment toxicity studies. AB - An automated water-renewal toxicity test system is described for exposing benthic invertebrates to whole sediments. The system will intermittently deliver laboratory or on-site water for overlying water replacement in sediment exposures. A range of cycle rates can be used to produce different volume additions of overlying water per day to exposure chambers. The system can be used with six different treatments and eight replicates per treatment producing 48 exposure chambers. Three formulated sediments with variable organic carbon (1.5%, 7.5%) and sand (14%, 63%) content were prepared to test the system exposing amphipods, Hyalella azteca and midges, Chironomus tentans in 10 day whole sediment tests. Intermittent water flow was used with a 90 min cycle time to create two volume additions of laboratory water per 24 h in exposure chambers (180 ml sediment, 320 ml water). Overlying water quality conditions, and survival and growth of both species were consistent and within acceptable limits for the testing requirements of the U.S. EPA guidelines for sediments with freshwater invertebrates. PMID- 12531305 TI - Cadmium and lead in tissues of Mallards (Anas platyrhynchos) and Wood Ducks (Aix sponsa) using the Illinois River (USA). AB - Cadmium and lead concentrations were determined in the tissues of Mallards and Wood Ducks collected from two waterfowl management areas along the Illinois River, USA, during the autumn and late winter of 1997-1998. Lead concentrations in livers of Mallards were lower than previously reported, and, along with those in a small sample of Wood Duck livers, were within background levels (<2.0 microg/g wet weight). Mean concentrations of cadmium in the kidneys of Wood Ducks utilizing the Illinois River were four times greater than in after-hatch-year Mallards, and 14 times greater than in hatch-year Mallards. Concentrations of cadmium in the kidneys of Wood Ducks were comparable with those of specimens dosed with cadmium or inhabiting contaminated areas in previous studies. Wood Ducks utilizing wetlands associated with the Illinois River, and presumably other portions of the lower Great Lakes region, may be chronically exposed to cadmium. PMID- 12531306 TI - Spatial and temporal analysis of second-generation anticoagulant rodenticide residues in polecats (Mustela putorius) from throughout their range in Britain, 1992-1999. AB - Polecats (Mustela putorius) in Britain are currently expanding their range eastwards from Wales to reoccupy central and eastern areas of England. Second generation anticoagulant rodenticides (SGARs), to which polecats are exposed by eating contaminated prey, are used more extensively in these central and eastern regions, leading to fears of increased exposure, and possible resultant mortality. We measured bromadiolone, difenacoum, flocoumafen and brodifacoum concentrations in the livers of 50 polecats from areas that included newly recolonised habitats and found that at least one SGAR was detected in the livers of 13 out of 37 (35.1%) male and 5 out of 13 (38.5%) female polecats. Difenacoum and bromadiolone were detected most frequently. We then combined these data with measurements on another 50 individuals from earlier studies to create a dataset for 100 polecats collected throughout the 1990s from across the whole of their current range. Using this dataset, we determined if there was any evidence that contamination in polecats had increased during the 1990s and whether animals from England were more contaminated than those from Wales, as might be expected given regional differences in the patterns of SGAR use. Overall, 31 of the 100 polecats analysed to date contained SGAR residues. The incidence was a little higher (40%) in animals that died between January and June and this probably better reflects the overall proportion of animals that are sub-lethally exposed. There was no statistically significant change during the 1990s in the proportion of polecats exposed to SGARs nor any evidence that greater use of SGARs in England resulted in more contamination of polecats. Contrary to expectation, the proportion of animals that contained difenacoum was marginally higher in Wales than elsewhere. PMID- 12531307 TI - The dissipation, distribution and fate of a branched 14C-nonylphenol isomer in lake water/sediment systems. AB - A single tertiary isomer which is believed to be one of the major branched isomers of the isomeric nonylphenol was synthesized for use in investigations on its metabolism and estrogenicity in aquatic organisms. The physico-chemical properties of the isomer were determined to enable the prediction of its behaviour in aquatic environments. From laboratory investigations on its dissipation and distribution in lake water, which are reported in this paper, it was found that it had a half-life of dissipation of 38.1 days and 20.1 days in an open lake water and in an open lake water/ sediment system, respectively, and to be rapidly partitioned in to sediment giving a high concentration factor of 1.76 after 28 days with an initial dose concentration of 2.52 ppm. The main dissipation route was found to occur through volatilization and co-distillation. The isomer was, however, found to be resistant to biodegradation in both the lake water and sediment, showing only a slight 9% loss (after 56 days) and 4.2% loss (after 28 days), of the 14C-residues in lake water and lake water/sediment systems, respectively, by microbial activity. Transformation to other more polar metabolites possibly by hydroxylation was also found to be minimal in both lake water and sediment samples after 14 days by HPLC analysis. After 7 days, only 2.25 and 7.4% transformation to a more polar metabolite was detected in lake water and sediment samples, respectively. PMID- 12531308 TI - Toxicity to freshwater organisms from oils and oil spill chemical treatments in laboratory microcosms. AB - Toxicity and temporal changes in toxicity of freshwater-marsh-microcosms containing South Louisiana Crude (SLC) or diesel fuel and treated with a cleaner or dispersant, were investigated using Chironomus tentans, Daphnia pulex, and Oryzias latipes. Bioassays used microcosm water (for D. pulex and O. latipes) or soil slurry (for C. tentans) taken 1,7, 31, and 186 days after treatment. SLC was less toxic than diesel, chemical additives enhanced oil toxicity, the dispersant was more toxic than the cleaner, and toxicities were greatly reduced by day 186. Toxicities were higher in the bioassay with the benthic species than in those with the two water-column species. A separate experiment showed that C. tentans' sensitivity was intermediate to that of Tubifex tubifex and Hyallela azteca. Freshwater organisms, especially benthic invertebrates, thus appear seriously effected by oil under the worst-case-scenario of our microcosms. Moreover, the cleaner and dispersant tested were poor response options under those conditions. PMID- 12531309 TI - Deriving freshwater quality criteria for 2,4-dichlorophenol for protection of aquatic life in China. AB - Freshwater quality criteria for 2,4-dichlorophenol (2,4-DCP) were developed with particular reference to the aquatic biota in China, and based on USEPA's guidelines. Acute toxicity tests were performed on nine different domestic species indigenous to China to determine 48-h LC50 and 96-h LC50 values for 2,4 DCP. In addition, 21 day survival-reproduction tests with Daphnia magna, 30-day embryo-larval tests with Carassius auratus, 60 day fry-juvenile test with Ctenopharyngodon idellus, 30 d early life stage tests with Bufo bufo gargarizans and 96 h growth inhibition tests with Scenedesms obliqaus were conducted, to estimate lower chronic limit (LCL) and upper chronic limit (UCL) values. The final acute value (FAV) was 2.49 mg/l 2,4-DCP. Acute-to-chronic ratios (ACR) ranged from 3.74 to 22.5. The final chronic value (FCV) and the final plant value (FPV) of 2.4-DCP were 0.212 mg/l and 7.07 mg/l respectively. Based on FAV, FCV, and FPV, a criteria maximum concentration (CMC) of 1.25 mg/l and a criterion continuous concentration (CCC) of 0.212 mg/l were derived. The results of this study provide useful data for deriving national or local water quality criteria for 2,4-DCP based on aquatic biota in China. PMID- 12531310 TI - A comparison of polycyclic aromatic hydrocarbon and petroleum hydrocarbon uptake by mussels (Perna viridis) and semi-permeable membrane devices (SPMDs) in Hong Kong coastal waters. AB - The ability of mussels (Perna viridis) and semi-permeable membrane devices (SPMDs) to accumulate polycyclic aromatic hydrocarbons (PAHs) and petroleum hydrocarbons (PHCs) from five sites in Hong Kong's coastal waters was compared. Mussels consistently had higher levels of contaminants, but their utility was limited at one highly polluted site due to mortality. Mussels and SPMDs ranked sites differently in terms of individual contaminant levels. Although SPMDs overcome many of the disadvantages of using living organisms to measure contaminants in marine waters, they cannot be used as "mimics" due to different PAH and PHC accumulation patterns. PMID- 12531311 TI - Combination of magnetic parameters: an efficient way to discriminate soil contamination sources (south France). AB - Biplots combining magnetic parameters allow to identification and differentiation different pollutant emission sources. A major problem in soil pollution is the characterization of the relative contributions of different anthropogenic particles sources. This paper demonstrates the efficiency of magnetic techniques to provide identification and differentiation of contaminating emission sources. About 100 soil samples were collected across a mixed agricultural and industrial area (Crau plain/Berre-Fos basin) in southern France. Nine soil profiles were realized. They are aligned along a transect, from the Mediterranean cost to the north. Measurements of initial magnetic susceptibility (chi) and remanent magnetization (ARM, IRM) have been carried out at room temperature. Several ratios of magnetic parameters were calculated and tested. Bivariate analyses allow to characterize different pollution sources and graphic results suggest three dominant contributions originated from road traffic, airport and steel industry. Moreover, magnetic grain-size discrimination between surface-soil samples and bottom-soil samples is obtained. An increase of hard magnetic components from topsoil towards the bottom of the profiles is evidenced. PMID- 12531312 TI - Characterization of contaminated soil and groundwater surrounding an illegal landfill (S. Giuliano, Venice, Italy) by principal component analysis and kriging. AB - The characterization of a hydrologically complex contaminated site bordering the lagoon of Venice (Italy) was undertaken by investigating soils and groundwaters affected by the chemical contaminants originated by the wastes dumped into an illegal landfill. Statistical tools such as principal components analysis and geostatistical techniques were applied to obtain the spatial distribution of chemical contaminants. Dissolved organic carbon (DOC), SO4(2-) and Cl- were used to trace the migration of the contaminants from the top soil to the underlying groundwaters. The chemical and hydrogeological available information was assembled to obtain the schematic of the conceptual model of the contaminated site capable to support the formulation of major exposure scenarios, which are also provided. PMID- 12531313 TI - A cross-taxa survey of organochlorine pesticide contamination in a Costa Rican wildland. AB - Amphibians, turtles, birds (mostly passerines) and mice collected from a conservation area in northwestern Costa Rica were analyzed for organochlorine (OC) pesticide contamination. Six of 39 amphibians (three of eight species), three of six turtles (two species), one of eight mice (one species) and 19 of 55 birds (five of seven species) contained OCs at levels up to 580 ng/g. The most frequently detected compound in 23 of 108 organisms was p,p'DDE. Dieldrin, delta BHC, heptachlor, p,p'DDD, and endosulfan II were each found in at least four organisms, while eight other OCs were found in at least one organism. The presence of OCs in taxa from the conservation area indicates the likelihood of long-distance transport of such compounds through the atmosphere. PMID- 12531314 TI - Total, dissolved, and bioavailable metals at Lake Texoma marinas. AB - Dissolved metals in water and total metals in sediments were measured at marina areas in Lake Texoma during June 1999 to October 2001, and October 2001, respectively. The metals most often found in the highest concentrations in marina water were Na and Ca, followed by Mg and K. Elevated Cu levels detected in lake water appeared to be associated with Cu based anti-fouling paint used on boats. Metal concentrations in sediment were much higher than in water. The relative order of the concentration in sediment was Ca > Al > Fe > K > Mg > Na. Elevated Cu level at specific locations appeared to be associated with local anthropogenic sources of boat repair activities. There were positive relationships between several metal elements in water and sediment. Metals in 16 sediments from lake marinas were extracted with a weak electrolyte solution [0.1 M Ca(NO3)2] to predict the bioavailability of metals. Among the five heavy metals studied (As, Cd, Cr, Cu and Zn), Cu was the most bioavailable in Lake Texoma marinas. PMID- 12531315 TI - Distribution of mercury in the aquatic environment at Almaden, Spain. AB - The world's largest mercury mine is placed at Almaden, Spain. However, there is a lack of information about the environmental impact of these mining activities in the ecosystem that surrounds this area. The aim of this article is to document the concentration of mercury in waters, sediments and bivalves of the aquatic system impacted by historic mine wastes. Simultaneously, a comprehensive study has been undertaken to characterise this hydrosystem and to determine the influence of some major physico-chemical parameters on the fate of mercury. Samplings were carried out for the last few years. Concentration of mercury in waters ranged from not detectable to 20 microg/l. For the sediments study, samples have been taken both from contaminated and non-contaminated sites within the basin. The regional background mercury concentration is higher than values typically cited for natural backgrounds. At exposed sites the mercury concentrations between 5 and 1000 microg/g were measured. These values are one to four order of magnitude greater than regional background levels. In the comparison between the results obtained at the present moment and those available for the 1974-1977 period, a general diminution of mercury levels is observed. Mercury concentrations in fresh water bivalves ranged between 1 and 4 microg/g (d.w.), with around 30% as monomethylmercury. In the discussion of the implications for risk assessment data available for other areas affected both for mine activities and mercuriferous belt are included. PMID- 12531316 TI - Total vs. internal element concentrations in Scots pine needles along a sulphur and metal pollution gradient. AB - Analysis of foliar elements is a commonly used method for studying tree nutrition and for monitoring the impacts of air pollutants on forest ecosystems. Interpretations based on the results of foliar element analysis may, however, be different in nutrition vs. monitoring studies. We studied the impacts of severe sulphur and metal (mainly Cu and Ni) pollution on the element concentrations (Al, Ca, Cu, Fe, K, Mg, Mn, Ni, P, Pb, S and Zn) in Scots pine (Pinus sylvestris L.) foliage along an airborne sulphur and metal pollution gradient. Emphasis was put on determining the contribution of air-borne particles that have accumulated on needle surfaces to the total foliage concentrations. A comparison of two soil extraction methods was carried out in order to obtain a reliable estimate of plant-available element concentrations in the soil. Element concentrations in the soil showed only a weak relationship with internal foliar concentrations. There were no clear differences between the total and internal needle S concentrations along the gradient, whereas at the plot closest to the metal smelter complex the total Cu concentrations in the youngest needles were 1.3-fold and Ni concentrations over 1.6-fold higher than the internal needle concentrations. Chloroform-extracted surface wax was found to have Ni and Cu concentrations of as high as 3000 and 600 microg/g of wax, respectively. Our results suggest that bioindicator studies (e.g. monitoring studies) may require different foliar analysis techniques from those used in studies on the nutritional status of trees. PMID- 12531317 TI - PCBs and organochlorine pesticides in blubber biopsies from free-ranging St. Lawrence River Estuary beluga whales (Delphinapterus leucas), 1994-1998. AB - For the first time, organochlorine (OC) contaminants were measured in blubber biopsies from free-ranging St. Lawrence River Estuary beluga whales (Delphinapterus leucas), to compare contaminant levels to those previously measured in dead stranded belugas. PCBs, DDTs, toxaphene and chlordane-related compounds were the major OC contaminants detected in 44 belugas biopsied in 1994 1998. SigmaPCB (the sum of 104 congeners) ranged from 2080 to 128,000 ng/g lipid in males (n=34; minimum estimated ages 8-22 years), and from 148 to 44,100 ng/g lipid in females (n=10; minimum estimated ages 7-22 years). The concentrations of PCBs and OC pesticides in the blubber of these whales overlapped those observed in stranded belugas from an earlier study, and demonstrated comparable age and sex-related trends. However, lower proportions of mirex, HCB, DDTs, and many of the highly chlorinated PCBs occurred in the biopsy samples compared to results for blubber from stranded carcasses. Most major OC compounds were present at lower concentrations in the biopsies, but this does not appear to be solely related to age differences between the two groups, or to emaciation in the stranded whales. Nor does it appear to be associated with the use of superficial biopsies, and the possible stratification of lipids and OCs in the blubber layer. Nevertheless, given these possible confounding factors, and the uncertainty in age estimates for the biopsied whales, the results point to the need for careful interpretation of biopsy results when comparing with data taken from the full depth of the blubber mantle in stranded whales. Taken together, results from both biopsied whales and previously studied stranded belugas indicate that PCB and OC pesticide contamination of St. Lawrence beluga whales may occur across a broader range of levels than previously thought, at least for males which formed the largest group in this study, possibly due to different degrees of dietary exposure. It also appears that measuring contaminant concentrations only in stranded whales, may overestimate OC levels in the population as a whole, especially for highly chlorinated OCs. PMID- 12531318 TI - The effects of soil amendments on heavy metal bioavailability in two contaminated Mediterranean soils. AB - Two heavy metal contaminated calcareous soils from the Mediterranean region of Spain were studied. One soil, from the province of Murcia, was characterised by very high total levels of Pb (1572 mg kg(-1)) and Zn (2602 mg kg(-1)), whilst the second, from Valencia, had elevated concentrations of Cu (72 mg kg(-1)) and Pb (190 mg kg(-1)). The effects of two contrasting organic amendments (fresh manure and mature compost) and the chelate ethylenediaminetetraacetic acid (EDTA) on soil fractionation of Cu, Fe, Mn, Pb and Zn, their uptake by plants and plant growth were determined. For Murcia soil, Brassica juncea (L.) Czern. was grown first, followed by radish (Raphanus sativus L.). For Valencia soil, Beta maritima L. was followed by radish. Bioavailability of metals was expressed in terms of concentrations extractable with 0.1 M CaCl2 or diethylenetriaminepentaacetic acid (DTPA). In the Murcia soil, heavy metal bioavailability was decreased more greatly by manure than by the highly-humified compost. EDTA (2 mmol kg(-1) soil) had only a limited effect on metal uptake by plants. The metal-solubilising effect of EDTA was shorter-lived in the less contaminated, more highly calcareous Valencia soil. When correlation coefficients were calculated for plant tissue and bioavailable metals, the clearest relationships were for Beta maritima and radish. PMID- 12531319 TI - Vaccines and Immunisation. Proceedings of the Third World Congress on Vaccines, and Immunisation. Opatija, Adriatic Riviera, Croatia. June 4-9, 2002. PMID- 12531320 TI - Towards the new global vaccinology era in prevention and control of diseases. PMID- 12531321 TI - Bioterrorism and the biotechnologist. PMID- 12531322 TI - New tendencies and strategies in international immunisation: GAVI and The Vaccine Fund. AB - It is now accepted that investment in health is an important contributor to poverty reduction; however failure to immunise the world's children with readily available life-saving vaccines results in more than 3 million premature deaths annually. The Vaccine Fund-a new structural tool with a clear mission "every child every where" and the Global Alliance for Vaccines and Immunization (GAVI)-a true alliance of all global stakeholders, are working together to build sustainability of immunisation and immunisation capacity in the world's poorest countries.Substantial funding for immunisation has been raised but more is needed to complete the task of fully immunising the world's poorest children. PMID- 12531323 TI - Vaccinology: past achievements, present roadblocks and future promises. AB - Of all the branches of modern medicine, vaccinology can claim to be the one that has contributed most to the relief of human misery and the spectacular increase in life expectancy in the last two centuries. It is the only science that has eradicated an infectious disease-smallpox-responsible for 8-20% of all deaths in several European countries in the 18th century. Other disabling and lethal diseases, like poliomyelitis and measles, are targeted for eradication. Currently, it is estimated that immunization saves the lives of 3 million children a year but 2 million more lives could be saved by existing vaccines. The success of vaccines in controlling and eliminating diseases has, paradoxically, been the cause of a revival of the anti-vaccination movement which in the absence, in developed countries, of many erstwhile common infectious diseases such as diphtheria, tetanus, polio, pertussis, measles, rubella and mumps has come to believe that vaccination is not only no longer necessary but is even dangerous. This is because it accepts, as "reactions", any untoward health event that occurs after administration of a vaccine. Most vaccine "reactions", therefore, appear to be more frequent than vaccine-preventable diseases. Public Health Authorities, aware of the great value of vaccines to society, are facing an uphill battle to get them accepted by a growing proportion of so-called educated minorities, thus endangering disease elimination. Other developments, in the last two decades, that have hampered vaccine usage have been the exploding costs of research, development and manufacture of new vaccines and the emphasis still placed on therapy in preference to prevention in medicine. This has led to the erroneous perception that vaccines are expensive although they are, in most cases, more cost-effective than the popular wait-see-treat approach. A favorable trend for vaccinology has been fueled by recent major breakthroughs in the sciences of immunology, molecular biology, genomics, proteomics, physico chemistry and computers that promise a bright future for prevention, not only of acute infectious diseases, but also treatment of conditions like chronic infections, allergy, auto-immune diseases and cancer where some malfunctioning of the immune system is thought to play a part. Vaccines are being made more user friendly by the development of combined vaccines and less painful and invasive inoculation techniques than the traditional syringe and needle. Recent new initiatives, like the Global Alliance on Vaccines and Immunization (GAVI),which are gathering new sources of funding for vaccination, should be beneficial for vaccinology. PMID- 12531324 TI - The global value of vaccination. AB - While most agree that vaccination is one of the most important public health practices, vaccines continue to be underused and undervalued, and vaccine preventable diseases remain a threat to world health. Perhaps one reason this gap remains is that decision-making generally is made on a vaccine-by-vaccine basis. There has been less attention to the value of vaccination in general. To more clearly identify this value, this paper reviews the cost-effectiveness literature and calculates the annual benefits of vaccination on a global scale. PMID- 12531325 TI - Recent progress in the development of vaccines for infants and children. AB - Infectious agents do not respect national or international boundaries. Attempts to prevent their spread, and the diseases which they cause, involve implementing vaccination as widely and as appropriately as possible. The principles of vaccination against infectious agents are now being applied to cancer and other non-infectious conditions. In order to understand where modern vaccinology is heading, it is necessary to first examine individual components. This brief overview examines the following: Streptococcus pneumoniae, Neisseria meningitidis, varicella zoster, measles, rotavirus, HIV, influenza, "emerging" viral infections and cancer. PMID- 12531326 TI - Two years into reverse vaccinology. AB - During the last century, several approaches have been used for the development of vaccines, going from the immunization with live-attenuated bacteria up to the formulation of the safer subunit vaccines. This conventional approach to vaccine development requires cultivation of the pathogen and its dissection using biochemical, immunological and microbiological methods. Although successful in several cases, this method is time-consuming and failed to provide a solution for many human pathogens. Now genomic approaches allow for the design of vaccines starting from the prediction of all antigens in silico, independently of their abundance and without the need to grow the microorganism in vitro. A new strategy, termed "Reverse Vaccinology", which has been successfully applied in the last few years, has revolutionized the approach to vaccine research. The Neisseria meningitidis serogroup B project, the first example of Reverse Vaccinology, as well as the application of this strategy to develop novel vaccines against other human pathogens are discussed. PMID- 12531327 TI - Novel approaches in the development of immunopotentiating reconstituted influenza virosomes as efficient antigen carrier systems. AB - One of the most recent major advances in vaccinology, and currently the only licensed alternative to the traditional alum-based adjuvants, are immunopotentiating reconstituted influenza virosomes. The flexibility of this novel antigen delivery system enables the safe and effective delivery of modern vaccine developments, such as viral glycoproteins, bacterial toxoids, inactivated virus, recombinant proteins, synthetic peptides and DNA-plasmids or polynucleotides. These novel vaccine antigens are being incorporated into immunopotentiating reconstituted influenza virosomes in an attempt to develop a variety of new viral, bacterial and parasitic vaccines. In addition, immunopotentiating reconstituted influenza virosomes are showing future potential in the targeting of specific cells, such as tumor cells, and novel routes of vaccine administration. PMID- 12531328 TI - Prospects and challenges for prophylactic and therapeutic HIV vaccines. AB - The best strategy for controlling the HIV pandemic remains the development of an efficacious prophylactic vaccine. Efficacy trials performed during this decade will yield information on the protective ability of first-generation vaccines. Several novel mixed-modality vaccines capable of inducing high-frequency CD8(+) T cell responses in macaques are being evaluated in humans. New hope has been raised with the prospect of therapeutic HIV vaccines. However, the development of HIV vaccines remains a formidable challenge to both the industry and the scientific community. PMID- 12531329 TI - Therapeutic vaccination for future management of HIV/AIDS. AB - The investigation of novel and innovative treatment approaches for long-term management of HIV-infection has intensified due to the growing number of infected individuals worldwide and the constraints of resistance, toxicity and inconvenience associated with lifelong therapy. Current treatment relies entirely on antiretroviral drugs targeting various stages of the life cycle of HIV, rather than on leveraging the immune system. However, the boosting of HIV-specific immune responses in chronic infection offers a vast potential for synergy with antiretroviral drugs, thereby contributing to durable control of viral replication. A novel immunotherapeutic agent that delivers plasmid DNA to dendritic cells after topical skin application is the first therapeutic vaccine that has demonstrated immunological and clinical benefit in chronically infected rhesus macaques in combination with antiretroviral therapy. PMID- 12531330 TI - V-1 Immunitor: oral therapeutic AIDS vaccine with prophylactic potential. AB - V-1 Immunitor (V1) is a therapeutic vaccine comprising pooled HIV antigens formulated into an oral pill. Recent V1 studies demonstrated body weight gain, increase in CD4 and CD8 cells, decrease in viral load, and improved survival of end-stage AIDS patients. The potential of V1 as a prophylactic vaccine has been evaluated in a phase II placebo-controlled trial on 35 volunteers. Twenty HIV negative volunteers who received V1 b.i.d. for 4 weeks had gained 28.2 and 17.5% in absolute CD4 (825 versus 1058; P=0.007) and CD8 (597 versus 702; P=0.013) cells, while lymphocytes in placebo group did not increase, suggesting that CD4 and CD8 counts may become an easily measurable immune correlate of the efficacy of AIDS vaccines. V1 does not appear to induce HIV-specific antibodies as orally administered immunogens usually produce cell-mediated but not systemic humoral response. V1 as a preventive oral vaccine targeting cellular and mucosal immunity deserves further evaluation. PMID- 12531331 TI - A Gag-Pol/Env-Rev SIV239 DNA vaccine improves CD4 counts, and reduce viral loads after pathogenic intrarectal SIV(mac)251 challenge in rhesus Macaques. AB - DNA vaccines are an important vaccine approach for many infectious diseases including human immunodeficiency virus (HIV). Recently, there have been exciting results reported for plasmid vaccination in pathogenic SHIV model systems. In these studies, plasmid vaccines supplemented by IL-2 Ig cytokine gene adjuvants or boosted by recombinant MVA vectors expressing relevant SIV and HIV antigens prevented CD4(+) T-cell loss and lowered viral loads following pathogenic challenge. However, similar results have not been reported in a direct pathogenic macaque challenge model. Here we report on a study of the ability of a multiplasmid SIV DNA vaccine in a pathogenic SIV251 rhesus mucosal challenge study. We observed that pGag/Pol+pEnv/Rev plasmid vaccines could not prevent SIV infection; however, vaccinated animals exhibited significant improvement in control of viral challenge compared to control animals. Furthermore, vaccinated animals exhibited protection against CD4(+) T-cell loss. PMID- 12531332 TI - Virus-like particle and DNA-based candidate AIDS vaccines. AB - Both humoral and cellular immune responses are critical for the control of HIV infection and replication. We have established systems for production of HIV and SIV virus-like particles containing high levels of viral Env proteins using the baculovirus expression system. Evaluation of immunogenicity showed that immunization with virus-like particles induced both cellular and neutralizing antibody responses. Furthermore, mucosal administration of virus-like particles effectively induced both mucosal and systemic immune responses. These results indicate that virus-like particles consisting of HIV structural proteins are an attractive vaccine platform for eliciting anti-viral immune responses, especially neutralizing antibody responses. We have also synthesized codon-optimized genes for HIV Env proteins and evaluated their immunogenicity. Combinations of virus like particle and DNA-based vaccination are promising for inducing strong cellular and neutralizing antibody responses against HIV. PMID- 12531333 TI - Immunogenicity of an HIV-1 gag DNA vaccine carried by attenuated Shigella. AB - The use of live attenuated invasive bacteria as a carrier for DNA-based vaccines has been reported recently. In this study, we used a Shigella flexneri serotype 2a rfbF mutant for immunization of a DNA vaccine coding for HIV-1 SF2 Gag. The recombinant bacterial vector delivered gag DNA to mammalian cells in vitro resulting in Gag protein expression, and was found to have a low level of pathogenicity among a number of Shigella cell spread defective mutants tested. Intranasal immunization of mice with live recombinant bacterial cells induced a gag-specific cellular immune response similar to that seen with i.m. injection of naked DNA. Importantly, a strong boosting effect was observed in mice primed with DNA, suggesting utility of bacterial vectors in prime-boost vaccination regimens. PMID- 12531334 TI - Induction of immune responses by DNA vaccines in large animals. AB - It is generally recognized that DNA vaccines are often less effective in large animals than in mice. One possible reason for this reduced effectiveness may be transfection deficiency and the low level of expression elicited by plasmid vectors in large animals. In our attempt to enhance transfection efficiency and, thereby, enhance immune responses, we employed a variety of methods inducing gene gun delivery or suppositories as delivery vehicles to mucosal surfaces, as well as electroporation for systemic immunization. To test these different systems, we used two different antigens-a membrane antigen from bovine herpesvirus glycoprotein (BHV-1) gD and a particulate antigen from hepatitis virus B. Gene gun and suppository delivery of BHV-1 gD to the vagina resulted in the induction of mucosal immunity not only in the vagina, but also at other mucosal surfaces. These data support the contention of a common mucosal immune system. In the case of electroporation, we were able to develop significant enhancement of gene expression following electroporation with surface electrodes (non-invasive electroporation) as well as invasive electroporation using single or six-needle electrodes. Various delivery systems such as bioject or needle delivery also influenced the immune response in both the presence and absence of electroporation. These studies also demonstrated that co-administration of plasmids coding for two different antigens (BHV-1 gD and hepatitis B surface antigen (HbsAg)) did not result in significant interference between the plasmids. These studies suggest that various combinations of delivery systems can enhance immunity to DNA-based vaccines and make them practical for administration of these vaccines in large animals. PMID- 12531335 TI - Progress in DNA vaccine for prophylaxis and therapy of hepatitis B. AB - Increasing lines of evidence suggest that DNA vaccine is of interest to fight chronic hepatitis B virus (HBV) infection. We used the Pekin duck infected by duck HBV (DHBV), closely related to the human virus, which is an attractive model allowing study of protective and therapeutic effectiveness of DNA vaccines against hepatitis B. Immunisation with a plasmid encoding the DHBV large (L) envelope protein induced a strong, specific, highly neutralising and long-lasting anti-preS humoral response in uninfected ducks. Importantly, maternal antibodies elicited by such DNA immunisation were vertically transmitted and protected progeny against viral challenge. Therapeutic immunisation of chronic DHBV-carrier ducks with this plasmid DNA led to the dramatic and sustained decrease in viral replication and even to clearance of intrahepatic viral covalently close circular DNA (cccDNA) pool in some animals. Our recent combination therapy data showed even a more pronounced antiviral effect of DNA vaccine to DHBV envelope protein when associated with antiviral drug (lamivudine) treatment. Therefore, DNA-based vaccine appears as a promising new approach for prophylaxis and therapy of hepatitis B. PMID- 12531336 TI - The rationale of a peptide-conjugate vaccine against measles. AB - The live-attenuated measles vaccine is poorly immunogenic in infants because of immune suppressive maternal antibodies and immaturity of the infant's immune system. Selected peptides corresponding to sequential, subdominant B cell epitopes of measles virus (MV) glycoproteins have been shown to induce neutralizing and protective antibodies even in the presence of whole virus antibodies. Similar to polysaccharide-conjugate vaccines, which are highly effective in infants a peptide-conjugate vaccine against measles is proposed. Such a vaccine induces carrier-specific T cells, avoiding measles-specific Th2 cells associated with the risk of atypical measles. This article discusses the rationale of such a strategy and its future potential. PMID- 12531337 TI - Mycobacterium bovis BCG-based vaccines against tuberculosis: novel developments. AB - Mycobacterium bovis Bacille Calmette-Guerin (BCG) is one of the most widely used vaccines. Modern techniques in genome manipulation allow the construction of recombinant (r)-BCG strains that can be employed as highly immunogenic vaccines against tuberculosis (TB) with an enhanced safety profile. In addition, the development of novel procedures to cultivate BCG will allow the large-scale production of future BCG-based vaccines. PMID- 12531338 TI - Development of surrogate markers for oral immunisation against Candida albicans. AB - We have developed an oral vaccine using the blastococcoid form of Candida albicans with a murine model of oral candidiasis with infection-resistant (BALB/c) and infection-prone (DBA/2) mice. Previous studies had demonstrated that enhanced clearance was linked to increased production of both Th1 and Th2 cytokines (IFN-gamma and IL-4, respectively) in regional lymph nodes, and increased secretion of IFN-gamma and NO in saliva. Subsequent study using oral Lactobacillus acidophilus (LAVRI-A1) induced a local cytokine pattern and enhanced clearance of intra-oral C. albicans similar to that noted in animals given killed candida vaccine. Thus, the experiment was repeated with a 2 month gap between completion of a vaccine course and challenge with live C. albicans. Compared to control mice, those given the oral vaccine had a 4 log reduction in colonisation on day 2, associated with twice the levels of secretion of IFN-gamma and IL-4 from the regional node cells, five times the level of nitric oxide in saliva, and suppression of NO production with MMLA induced a 2 log increase in oral colonisation. Thus while both IFN-gamma and IL-4 appear to contribute to clearance of oral C. albicans, IL-4 appears to act through a paracrine enhancement of NO production. Studies in man are in progress to define similar surrogate markers of immune protection, prior to oral vaccine trial. PMID- 12531339 TI - Experience with registered mucosal vaccines. AB - Most pathogens gain access to their host through mucosal surfaces. It is therefore desirable to develop vaccination strategies that lead to mucosal immune responses. Ideally, a vaccine should be administered mucosally in order to elicit mucosal protection. Several attenuated live viral and bacterial pathogens are registered as oral vaccines for human use, including the oral polio vaccine (Sabin) as well as attenuated strains of Salmonella typhi and Vibrio cholerae. These attenuated bacterial live vaccines-S. typhi Ty21a as well as V. cholerae CVD 103-HgR-are employed as vaccines against typhoid and cholera, respectively. In this manuscript, we review the immune responses that are induced by these vaccines, with a focus on mucosal immunity. PMID- 12531340 TI - Impact of universal vaccination programmes on the epidemiology of hepatitis B: 10 years of experience in Italy. AB - Ten years have elapsed since routine vaccination of infants and of 12-year-old adolescent was implemented in Italy. In this period, evidence has accumulated on the epidemiological impact of universal immunisation. Coverage is on average >90% and is >or=95% in many areas of the country. Incidence of acute hepatitis B, that was already declining before 1991, was further decreased by routine vaccination programmes. This is particularly evident in adolescents and young adults (cohorts involved by mandatory vaccination), while incidence shows little changes in older subjects according to data of the last years. Prevalence of hepatitis B virus (HBV) markers detected by sero-epidemiological studies on anonymous sera confirms both the very high coverage with hepatitis B vaccination and the virtual absence of chronic HBsAg carriers in cohorts involved by routine vaccination programmes. The system of passive surveillance on adverse events following hepatitis B vaccination supports the excellent safety record of hepatitis B vaccines. In a hyperendemic area of Southern Italy, where a pilot programme was firstly implemented, it was also possible to document the decline of the involvement of hepatitis B in chronic liver pathologies (from 48% in 1982 to 18% in 1997). If coverage rates are maintained at the present levels, elimination of HBV transmission in Italy may be envisaged in few decades. PMID- 12531342 TI - Vaccination strategies against hepatitis A in southern Europe. AB - Hepatitis A virus (HAV) infection is decreasing in southern European countries, where epidemiological conditions vary among regions depending on the social and health-care system development. In high endemic settings, HAV infection has not heavy social and economic weight while in countries with a moderate/low degree of endemia there is a call for targeted vaccination policy. In countries, like Spain and in Italy, where several studies confirm an increase in susceptible adults, vaccination strategies have been applied and recommendations have been published about hepatitis A prevention. Universal hepatitis A immunization seems economically unattractive and most evidences for targeted vaccination have not yet been sufficiently investigated. Vaccine should be used to protect travellers to countries where HAV infection is a major risk and in preventing secondary cases and outbreaks. PMID- 12531341 TI - Primary prevention of hepatitis C virus infection. AB - Viral hepatitis type C is a worldwide public health problem of major concern. Waiting for a safe and effective vaccine able to confer protection to susceptible individuals, public health challenges for controlling hepatitis C require implementation of primary prevention measures that reduce risks of acquiring/transmitting HCV infection. Screening for safe blood and blood products, use of disposable syringes and needles and of universal precautions have dramatically reduced risk of infection in medical setting. Health education, counselling and testing of individuals at risk provide opportunities for controlling HCV infection. PMID- 12531343 TI - Effectiveness of a mass hepatitis A vaccination program in preadolescents. AB - A program of mass hepatitis A+B vaccination in preadolescents in schools was begun in the Catalonia in the last quarter of 1998. This study investigated the impact of the program by comparing the incidence of hepatitis A in vaccinated and unvaccinated cohort. The greatest reduction of the incidence rate of hepatitis A was observed in the 10-14 years age group, from 10.3 per 100000 persons-year in the period 1996-1998 to 1.8 per 100000 persons-year in the period 1999-2001. The global incidence decreased from 6.2 to 2.6 per 100000 persons-year. After analysis of cases occurring in the vaccinated and non vaccinated cohort, the effectiveness of the vaccination program was estimated at 97.0% (95% CI: 78.6 99.6). PMID- 12531344 TI - Costs and benefits of polio eradication: a long-run global perspective. AB - Pre-vaccination polio incidence rates in USA and Italy were used to predict the cases that would have occurred in the world for the years 1970-2050 in the absence of immunization. Globally, polio program will cost about US dollars 67 billion if vaccination is discontinued after 2010. The medical care cost savings achieved will be more than US dollars 128 billions, implying that polio eradication activities actually pay for itself in the longer run. In addition to the cost savings, the program will prevent 855000 deaths, 4 million paralysis cases and 40 million disability adjusted life years (DALYs) over the years 1970 2050. PMID- 12531345 TI - The challenge to provide affordable rabies post-exposure treatment. AB - Treatment with Semple or suckling mouse brain rabies vaccines persists in many countries of Asia, Africa and South America. Its replacement depends on the immediate accessibility of effective affordable alternative treatment with tissue culture vaccines (TCVs). The use of the expensive European TCVs has been possible in Asia by means of economical intradermal (ID) post-exposure vaccine regimens. Implementation of this effective economical treatment has been delayed by the complexity and inconvenience of the regimens, and the reluctance to change prophylaxis against a fatal disease. Up to now, the ID regimens have been used only where passive immunisation with rabies immune globulin (RIG) is usually available. Rabies deaths despite optimal vaccine treatment have been attributed to lack of RIG. The ID regimens might soon be promoted in areas where RIG is not even available for severe exposure. It is therefore vital that economical vaccine regimens should be used which induce protective immunity rapidly. Improvements in rabies pet in developing countries could be made by: (i). publicising the urgency and efficacy of wound cleaning; (ii). facilitating the replacement of nervous tissue vaccines by economical ID treatment with TCVs; (iii). using an ID regimen with a large dose of vaccine on the first day of treatment especially when no RIG is available; and (iv). promoting pre-exposure prophylaxis to eliminate the need for RIG and provide better rabies prophylaxis. PMID- 12531346 TI - Human rabies prophylactics: the French experience. AB - In 1968, fox rabies was introduced on the French territory, in the Moselle department and from that time, spread southwards and westwards from the French German border at the speed of 40 km per year. Consequently, a program aimed at controlling and eradicating the disease was carried out. Collaboration between human and veterinary medicine has been the key of the success of this program. In 2001, rabies in terrestrial animals was eradicated from France, while no indigenous human rabies case had been reported. Meanwhile, post-exposure treatments (PET) had been closely monitored. Data on rabies cases in animals, rabies cases in humans, PET, surveillance of exposures to baits and oral vaccines for the wild fauna, and exposures outside the French territory will be successively considered and discussed. PMID- 12531347 TI - Evaluation of the national immunisation programme in the Netherlands: immunity to diphtheria, tetanus, poliomyelitis, measles, mumps, rubella and Haemophilus influenzae type b. AB - The immunity to vaccine-preventable diseases included in the Dutch immunisation programme in the general population and among orthodox reformed individuals who refuse vaccination was assessed. The programme induces good protection. However, a large proportion of adults lacks diphtheria and tetanus immunity. Measles, mumps and rubella seroprevalence was somewhat lower among vaccinated compared to unvaccinated cohorts. The prevalence of HibPS antibodies declined during 2.5 years after the fourth vaccination. However, protection occurs also by memory immunity. Herd immunity is sufficient among the general population, but not among orthodox reformed individuals. Immunosurveillance is an efficient way to evaluate the effects of immunisation programmes and identify risk groups for infection. PMID- 12531348 TI - Options for improvement of the Dutch measles vaccination schedule. AB - We investigated which vaccination schedule gives best protection to the vaccinating population, in case of a measles epidemic in pockets of unvaccinated individuals. We explored the effect of an additional measles vaccination (at 6 or 9 months), advancing the first measles-mumps-rubella (MMR) vaccination from 14 to 11 months, and advancing the second MMR from 9 to 4 years. Measures of protection among vaccinees (percentage of susceptibles, number of reported cases, percentage of lifetime spent susceptible) were estimated with a mathematical model of the impact of antibody level on seroconversion and immunity. Advancing the age of second MMR vaccination prevents considerably more cases among vaccinees than an extra early measles vaccination or advancing the age of first MMR vaccination. PMID- 12531349 TI - Impact of mass vaccination with polysaccharide conjugate vaccine against serogroup C meningococcal disease in Spain. AB - During the fourth quarter of 1997, a vaccination campaign using the meningococcal C polysaccharide vaccine was carried out in 14 autonomous regions of Spain. The remaining three regions did not participate. In the last quarter of the year 2000, a mass vaccination campaign using the meningococcal C conjugated vaccine was carried out in all regions. In the year 2001 the incidence decreased in all regions, although the decrease was greater in regions that did not vaccinate in 1997. In contrast, case fatality rates did not decrease. During 2001, the incidence rate of meningococcal C disease was still lower (0.32 per 100000 persons-year) in the regions that vaccinated in 1997 with the polysaccharide vaccine than in those that did not (0.64 per 100000 persons-year). PMID- 12531350 TI - Dramatic decline of serogroup C meningococcal disease in Catalonia (Spain) after a mass vaccination campaign with meningococcal C conjugated vaccine. AB - In the last quarter of the year 2000, the meningococcal C conjugated vaccine was incorporated into the routine vaccination schedule in Catalonia (at 2, 4 and 6 months). In addition a vaccination campaign was carried out in children <6 years of age, with a coverage of 96.2%. The effectiveness of the vaccination in this age group during 2001 and the first 28 weeks of 2002 was 100% (94.27-100%). A vaccination campaign has been carried out in 6-19-year olds during 2001 and 2002, with a coverage rate of 23.5% in the year 2001. In this age group a reduction in disease incidence was seen in the 2000-2001 season, but not in the following one. PMID- 12531351 TI - Serum bactericidal activity correlates with the vaccine efficacy of outer membrane vesicle vaccines against Neisseria meningitidis serogroup B disease. AB - For evaluation of serum bactericidal activity (SBA) as surrogate for the efficacy of outer membrane vesicle (OMV) vaccines against Neisseria meningitidis serogroup B disease, we have reanalyzed data from a randomized double blind placebo controlled efficacy trial involving 172000 secondary school students (aged 13-14 years) in Norway (1988-1991). A cohort of the efficacy trial consisting of 880 individuals was selected for immunogenicity studies. An efficacy of 87% was calculated for a 10-month observation period. However, after an observation period of 29 months, the estimated efficacy against group B disease induced by vaccination was 57%. The immunogenicity study showed that the SBA geometric mean titer (GMT) for the vaccinees was 2.4 before vaccination and 19.0 six weeks after the second vaccine dose. One year after vaccination the GMT was reduced to 2.8. A separate three-dose study with 304 adolescents showed that with a third dose at 10 months after the second dose (i.e. when cases of disease started to appear) a strong booster response was induced. Ten months after the second dose the SBA was reduced to near pre-immunization level. Following the third dose the SBA geometric mean titer of 2.7 increased to 62.3. One year after the third dose, the GMT was markedly higher than 6 weeks after the second dose (12.6 versus 8.8). Thus, protection after vaccination corresponds with the level of SBA. In order to reach lasting protective levels of SBA in a population, three vaccine doses are probably required. Measurements of SBA are likely to be useful for evaluating various upcoming formulations and improvements of immunization regimens for OMV vaccines. PMID- 12531352 TI - Clinical evaluation of a polygeline-free tick-borne encephalitis vaccine for adolescents and adults. AB - OBJECTIVE: To evaluate immunogenicity and safety of a polygeline-free tick-born encephalitis (TBE) vaccine in a clinical program. METHOD: A total of 3118 subjects aged 12-76 years were enrolled in three clinical trials. The clinical studies were conducted in 15 centers in three European countries. Evidence of neutralizing TBE antibodies was used as surrogate parameter for efficacy assessment. RESULTS: All subjects analyzed achieved levels of TBE antibodies postimmunization to fulfill the definition of seroconversion or a four-fold increase. The new TBE vaccine appeared to be well tolerated by subjects. Only very few febrile reactions, mainly 38.5 degrees C were reported. No serious or unexpected adverse events related to vaccination were reported. CONCLUSION: These successful results in terms of both immunogenicity and safety indicate that the TBE vaccination with this polygeline-free TBE vaccine can be used safely in adolescents and adults. PMID- 12531353 TI - Tolerability of modified tick-borne encephalitis vaccine FSME-IMMUN "NEW" in children: results of post-marketing surveillance. AB - A new, highly purified, inactivated tick-borne encephalitis (TBE) vaccine FSME IMMUN "NEW" has been developed by Baxter using a production virus seed derived from chick embryo cells instead of mouse brain. In clinical trials, the vaccine was shown to be highly immunogenic and well tolerated in adults and children. Following licensure in 2001, the tolerability of half the adult dose of FSME IMMUN "NEW" (1.2 microg antigen/0.25 ml) was investigated in a post-marketing surveillance in 1899 children aged 6 months to 12 years. Rectal body temperature was measured daily for 3 days after the first vaccination. An overall fever rate of 20.3% (95% CI=18.5; 22%) was observed, which was mostly mild in nature (>38.0 to 0.05). To better determine the usefulness of our in vivo methods to identify MHC class I-binding epitopes, four epitopes from the outer capsid VP7 rotavirus protein determined in ex vivo assays were evaluated for their protective efficacies and two were found to be partially protective. Together, these studies show that functional mapping is useful in locating epitopes that are relevant to the development of subunit rotavirus vaccines. PMID- 12531357 TI - Transcriptome-based antigen identification for Neisseria meningitidis. AB - The identification of suitable antigens is crucial to successful vaccine development based on subunit approaches. While many methods exist for the identification of vaccine candidates which are surface-exposed or secreted, immunogenic and conserved, contain B and T cell epitopes, most of these have a major drawback: they do not yield any information on whether the antigen is indeed expressed by the pathogen during infection. However, DNA microarrays offer a novel tool for the investigation of the transcriptional activity of all genes of a pathogenic microorganism under in vivo conditions. Employing whole genome DNA microarrays, we have analyzed the transcriptome of Neisseria meningitidis serogroup B bacteria during different stages of infection, i.e. exposure to human serum and the interaction with human epithelial and endothelial cells. Combined with data derived from genome-based approaches (such as reverse vaccinology) and immunogenicity studies, this novel transcriptome-based antigen identification should reveal ideal vaccine candidates against serogroup B meningococcal infection. PMID- 12531358 TI - Immunity under the skin: potential application for topical delivery of vaccines. AB - With the technological advances in biomedical sciences and the better understanding of how the immune system works, new immunisation strategies and vaccine delivery options, such sprays, patches, and edible formulations have been developed. This has opened up the possibility of administering vaccines without the use of needles and syringes. Already topical immunisation is a reality and it has the potential to make vaccine delivery more equitable, safer, and efficient. Furthermore, it would increase the rate of vaccine compliance and greatly facilitate the successful implementation of worldwide mass vaccination campaigns against infectious diseases. This review gives a brief account of the latest developments of application of candidate vaccine antigens onto bare skin and describes some of our recent observations using peptide and glycoconjugate vaccines as immunogens. PMID- 12531359 TI - Tumor antigen-specific T cells and cancer immunotherapy: current issues and future prospects. AB - The critical role of antigen-specific T cells in the eradication of cancer has been demonstrated in numerous animal models. Data compiled from both in vitro systems and human clinical trials indicate that T cells can be identified that recognize antigenic fragments derived from gene products expressed by tumors. Nonetheless, results from clinical trials have been for the most part disappointing, since vaccine protocols designed to elicit anti-tumor T cell activity have, in the majority of cases, failed to result in tumor eradication and enhanced patient survival. The focus of this review article is to summarize the current status of antigen-specific tumor immunotherapy and provide insight into potential future strategies for the successful activation of T cells for the immunotherapy of cancer. PMID- 12531360 TI - Dendritic cell (DC) based therapy for cervical cancer: use of DC pulsed with tumour lysate and matured with a novel synthetic clinically non-toxic double stranded RNA analogue poly [I]:poly [C(12)U] (Ampligen R). AB - Human papilloma virus (HPV) found in 99.7% of cervical cancers represents an attractive immunotherapeutic target for novel adjuvant dendritic cell (DC) immunotherapy. DC primed with HPV antigens have been shown to be capable of inducing CTL responses powerful enough to eradicate established murine tumours expressing HPV16 antigen. The use of tumour lysate has been found to be an effective means of priming DC with tumour associated antigens in animal models and in clinical trials leading to significant anti-tumour responses. Autologous DC primed with sonicated HPV expressing tumour lysate have been shown to be capable of inducing HPV specific classes I and II T-cell immunity in a pilot clinical study.Synthetic double stranded polyribonucleotides are effective in vitro activation/maturation agents capable of inducing a stable mature DC phenotype producing high levels of IL12. However, the prototype polymer poly [I]:poly [G] has proved to be clinically toxic. Preliminary in vitro data have demonstrated that a novel clinically non-toxic analogue polymer poly [I]:poly [C(12)U] (Ampligen R) can effectively induce in vitro maturation of human monocyte derived DC with sustained bioactive IL12 production. Human monocyte derived DC primed with tumour lysate and matured with synthetic dsRNA may therefore offer an effective way of optimising Th1 specific anti-cancer T-cell responses in cancer patients. This strategy is currently being tested in a clinical trial in patients with cervical cancer. PMID- 12531361 TI - Anti-cancer therapy using dendritic cells and apoptotic tumour cells: pre clinical data in human mesothelioma and acute myeloid leukaemia. AB - We have recently reported in an experimental model, that treatments based on the injections of dendritic cells which had phagocytosed apoptotic bodies derived from tumour cells were particularly effective in the cure of tumour-bearing animals. We proposed that systems using processing and presentation of antigenic molecules from antigen-presenting cells primed with apoptotic bodies can offer new opportunities in anti-cancer treatment. We first established the technical conditions for purification, characterisation and production of tumour cells isolated from fresh pleural liquid or blood. Then we compared efficacy of different apoptotic inducers agents on the cancer cells in culture. The apoptotic tumour cells were purified, characterised and maintained in coculture with monocytes-derived immature dendritic cells. We subsequently investigated the effect of the maturation process on phagocytosis of apoptotic bodies. We have shown that whatever the nature of the apoptotic cells they are phagocytosed by the dendritic cells which were efficiently matured using the combination of TNFalpha+Poly I:C. Furthermore, we demonstrated that the generation of the mature dendritic cells pulsed with apoptotic tumour cells, successfully generated CD4(+) (Th1) and CD8(+) (CTL) cells. All the experimental procedures that we have used were developed with clinical use in mind, using Good Manufacturing Products. We are presently investigating the feasibility of such a "vaccine" for the treatment of asbestos mesothelioma or acute myeloid leukaemia. PMID- 12531362 TI - Contribution of dead cells to the immunogenicity of an autologous, hapten modified melanoma vaccine. AB - We have reported that treatment of melanoma patients with a vaccine consisting of autologous tumor cells modified with the hapten, dinitrophenyl (DNP), induced delayed-type hypersensitivity (DTH) to autologous, unmodified tumor cells. Moreover, this response was a significant and independent predictor of survival. We analyzed the vaccines prepared for 284 patients who were treated following resection of regional or distant metastases to determine whether the dose and composition correlated with immunological response. Regression analysis showed no significant association between the magnitude of this DTH response and the number of live (trypan blue-excluding) melanoma cells per dose. In fact, vaccines containing higher numbers or higher proportions of dead, but intact, tumor cells induced larger DTH responses to autologous unmodified tumor. The observation that dead tumor cells are immunogenic may be applicable to other cellular human cancer vaccines and underscores the need for applying pharmacological principles to cancer immunotherapy. PMID- 12531363 TI - Towards new immunotherapies: targeting recombinant cytokines to the immune system using live attenuated Salmonella. AB - We have used Salmonella as a delivery system for eukaryotic expression plasmids encoding cytokines, and assessed its capacity to modulate immune responses in different experimental models. Plasmids encoding mouse IL-4 and IL-18 under cytomegalovirus promoter were constructed and transformed into live attenuated Salmonella enterica serovar Typhi strain CVD 908-htrA, and Salmonella enterica serovar Typhimurium strain SL3261. We have shown that systemic as well as mucosal immunization with such constructs can influence the antibody and cytokine responses to the Salmonella carrier and to co-administered bystander antigens, as well as the specific immune response elicited during a parasitic infection. Further, we have shown that oral cytokine-therapy using Salmonella as gene vector induce antitumoral effect as demonstrated by extended survival time in melanoma bearing mice. This approach may be particularly suited for the development of new immunotherapies with applications in parasitic infections and cancer, were alterations of the host's immune responses are usually found, and therapy-induced modulation of the immune response is likely to be required. PMID- 12531364 TI - Vaccine antigen production in transgenic plants: strategies, gene constructs and perspectives. AB - Stable integration of a gene into the plant nuclear or chloroplast genome can transform higher plants (e.g. tobacco, potato, tomato, banana) into bioreactors for the production of subunit vaccines for oral or parental administration. This can also be achieved by using recombinant plant viruses as transient expression vectors in infected plants. The use of plant-derived vaccines may overcome some of the major problems encountered with traditional vaccination against infectious diseases, autoimmune diseases and tumours. They also offer a convenient tool against the threat of bio-terrorism. State of the art, experimental strategies, safety and perspectives are discussed in this article. PMID- 12531365 TI - Targeting of plant-derived vaccine antigens to immunoresponsive mucosal sites. AB - Most pathogenic microorganisms enter their host via the mucosal surfaces lining the digestive, respiratory and urino-reproductive tracts of the body. The most efficient means of protecting these surfaces is through mucosal immunization. Transgenic plants are safe and inexpensive vehicles to produce and mucosally deliver protective antigens. However, the application of this technology is limited by the poor response of the immune system to non-particulate, subunit vaccines. Co-delivery of therapeutic proteins with targeting proteins, such as the B subunit of the Escherichia coli heat labile enterotoxin (LTB), could increase the effectiveness of such antigens. PMID- 12531366 TI - Corn as a production system for human and animal vaccines. AB - The synthesis of selected antigens in plants and their oral delivery has great potential for reducing the costs of vaccine production and administration. The application of this technology requires antigen concentrations in final plant material to be uniform to ensure consistent dosing. In addition, antigen levels should be such as to allow the volume of each dose, containing a set amount of antigen, to be practical for oral delivery. Here, we demonstrate that the Lt-B protein of enterotoxigenic E. coli is evenly distributed in defatted corn germ prepared from transgenic grain. Furthermore, the choice of sub-cellular location for Lt-B affects accumulation of the protein in excess of four orders of magnitude. PMID- 12531367 TI - Immunogenic measles antigens expressed in plants: role as an edible vaccine for adults. AB - Vaccine-induced immunity against measles is less robust than natural immunity. Waning of immunity in vaccines may eventually require a revaccination of adults. Measles antigens expressed in plants have been shown to be antigenic and immunogenic both after invasive and oral vaccination. Strategies for the vaccination of adults, the potential of an oral measles vaccine produced in edible plants and the design of suitable antigens are discussed. PMID- 12531368 TI - Production of antibodies in plants and their use for global health. AB - Recombinant antibodies can be used to diagnose, treat and prevent disease by exploiting their specific antigen-binding activities. A large number of drugs currently in development are recombinant antibodies and most of these are produced in cultured rodent cells. Although such cells produce authentic functional products, they are expensive, difficult to scale-up and may contain human pathogens. Plants represent a cost-effective, convenient and safe alternative production system and are slowly gaining acceptance. Five plant derived therapeutic recombinant antibodies (plantibodies) are undergoing clinical evaluation, three of which can be used as prophylactics. PMID- 12531369 TI - Trauma training and the military. PMID- 12531370 TI - Adopting the pre-hospital index for interfacility helicopter transport: a proposal. AB - BACKGROUND: Interfacility helicopter transport is expensive without proven outcome benefit in trauma patients. Our objectives were to determine the fastest method of rural to urban interfacility transport, and develop a triage tool to identify patients most in need of rapid transport. METHODS: Retrospective cohort study. Adults ISS > or = 12 transported from January 1996 to December 1998. Transport time variables were compared between geographical zones. A pre transport index (PTI) identified two patient cohorts in which outcome was assessed. RESULTS: Air ambulance was faster than ground transport, with helicopter overall superior to fixed-wing (< 225 km range). Seventy-two percent of patients with PTI < 4 (n = 196) had no outcome indicating severe injury versus 29% of the PTI > or = 4 cohort (n=151). Mortality for PTI<4 was 1.4% versus 22% for PTI > or = 4. CONCLUSION: Interfacility helicopter transport of severely injured rural trauma patients was the overall fastest method within a 225 km range. PTI > 4 identifies patients most in need of this fast but expensive method of transport. PMID- 12531371 TI - The effects of air ambulance admissions on orthopaedic trauma services in a single hospital. AB - Emergency air ambulance admissions to the authors' hospital have increased five fold from 18 in 1992-1993 to 92 in 1998-1999. The service implications for hospitals receiving air ambulance admissions is unknown. AIM: An audit/analysis of all emergency air ambulance admissions to the authors' hospital between August 1998-July 1999. METHOD: Admissions identified from computer records. The case notes were then retrospectively analysed. Data was collected on accident location, admitting specialty, number of orthopaedic procedures and their operative time, and length of inpatient stay. RESULTS: There were 92 patients brought in by air ambulance. Hospital notes were available for 82 and a further 8 had A&E case notes only available for analysis. A total of 34 (38%) were admitted under the orthopaedic surgeons, 28 (31%) under the neurosurgeons, 10 (11%) under the plastic surgeons, 2 (2%) under the general surgeons and 2 (2%) under the physicians. Nine patients were discharged home from the A&E department, two died in A&E and five were transferred to other hospitals. Four of the neurosurgical patients needed orthopaedic input. The admitted patients required 122 orthopaedic procedures taking 120 h of theatre time. The total orthopaedic inpatient stay was 628 days, of which 145 days were in the intensive care or high dependency units. A total of 28 (80%) of the orthopaedic patients came from outside the hospital's catchment area. CONCLUSIONS: Admissions by air ambulance place a high demand on orthopaedic services and often originate from outside the hospital's catchment area. Additional resources may be required by hospitals receiving trauma patients by this route. PMID- 12531372 TI - Psychological care in trauma patients. AB - The clinician manages trauma patients in the emergency room, operation theatre, intensive care unit and trauma ward with an endeavour to provide best possible treatment for physical injuries. At the same time, it is equally important to give adequate attention to behavioural and psychological aspects associated with the event. Knowledge of the predisposing factors and their management helps the clinician to prevent or manage these psychological problems. Various causes of psychological disturbances in trauma patients have been highlighted. These include pain, the sudden and unexpected nature of events and the procedures and interventions necessary to resuscitate and stabilise the patient. The ICU and trauma ward environment, sleep and sensory deprivation, impact of injury on CNS, medications and associated pre-morbid conditions are also significant factors. Specific problems that concern the traumatised patients are helplessness, humiliation, threat to body image and mental symptoms. The patients react to these stressors by various defence mechanisms like conservation withdrawal, denial, regression, anger, anxiety and depression. Some of them develop delirium or even more severe problems like acute stress disorder or post-traumatic stress disorder. Physical, pharmacological or psychological interventions can be performed to prevent or minimise these problems in trauma patients. These include adequate pain relief, prevention of sensory and sleep deprivation, providing familiar surroundings, careful explanations and reassurance to the patient, psychotherapy and pharmacological treatment whenever required. PMID- 12531373 TI - Sexual dimorphism in trauma? A retrospective evaluation of outcome. AB - PURPOSE: Multiple studies have demonstrated a heightened immune response in female animals subjected to trauma-hemorrhage models and have implied a subsequent survival advantage. PROCEDURES: A retrospective review of outcome in 15,170 trauma admissions over a 5-year-period (1993-1997) at a level 1-trauma center was performed. A comparison of outcome by gender, age, injury severity score (ISS), mechanism of injury, location of injury (AIS), and length of hospitalization (intensive care unit and total hospitalization) was performed. FINDINGS: There were 12,456 male and 2714 female patients included in the study. Overall survival rates (male = 90.2%, female = 90.8%) and survival of serious (ISS > or = 15) trauma (male = 63.5%, female = 60.5%) were not statistically different. Logistic regression analysis identified age, mechanism and ISS as factors associated with survival. CONCLUSION: Retrospective evaluation of our trauma population failed to show a difference in outcome between male and female trauma patients. Age, mechanism and severity of injury-but not gender-were identified as factors influencing survival. PMID- 12531374 TI - Pain as a presenting feature of acute abdomen in spinal injuries. AB - The diagnosis of acute abdomen can be difficult in patients with spinal injuries. We reviewed all the 1039 case records of patients admitted with spinal injuries to the Queen Elizabeth National Spinal Injuries Unit, Glasgow over a 7-year period and found 5 (0.48%) cases of acute abdomen that required surgical intervention and were not caused by original injury. Their presenting signs and symptoms were analysed. Pain was found to be an unreliable symptom in these patients. PMID- 12531375 TI - Self-inflicted abdominal stab wounds. AB - BACKGROUND: Self-inflicted abdominal stab wounds (ASWs) are uncommon. The present study aims to characterize the clinical profile of this unique group of psychiatric-surgical patients. METHODS: A retrospective review of 23 patients with intentional self-inflicted ASWs at two urban level I trauma centres during a 10-year period. RESULTS: Most patients were males (70%), ages ranging from 21 to 82 years (mean 40 years). Seventy-four percent of patients had a previous psychiatric history and prior suicide attempts were common (41%). Half of the patients had a positive admission drug or alcohol screen. Hypotension (systolic blood pressure (SBP) < 90 mmHg) was present in only two patients. Mean injury severity, revised trauma and Glasgow coma scores were 5.8, 7.7 and 14.5, respectively. The most commonly used instrument was a knife (87%). There were 1.5 external wounds per patient located primarily in the right upper quadrant (40%) and right lower quadrant (23%). These were associated with extra-abdominal wounds in 22% of cases. Local wound exploration was positive in 15 patients (65%), who all underwent laparotomy. Injured intra-abdominal or retroperitoneal organs were identified in 10 patients and included the stomach, duodenum, small bowel, colon, mesentery, inferior vena cava (IVC) and psoas muscle with a mean of 1.7 injuries per patient. Wound infection was the only post-operative complication (two patients). All eight patients with a negative local wound exploration were observed without complication. Seventy percent of patients were ultimately transferred to a psychiatric ward with a mean length of stay on the surgical service of 8 days. Only one patient died during admission due to metastatic malignant disease. CONCLUSION: Self-inflicted ASWs in suicidal patients can induce significant although most likely non-lethal abdominal and retroperitoneal injuries. This observation should guide the trauma surgeon when treating such patients. PMID- 12531376 TI - The morbidity of penetrating colon injury. AB - The purpose of this study is to define the current morbidity and mortality associated with penetrating colon injury and to determine the impact of management strategy on outcome. METHODS: A retrospective review was performed of all penetrating colon injuries managed at a level I trauma center (1990-2000), n=186. Stepwise logistic regression was used to determine the independent predictors for colostomy and morbidity following colon injury. RESULTS: Fifty three percent of the patients were managed with primary repair or anastomosis while 47% received a colostomy. Independent predictors of colostomy included gunshot wound (GSW), degree of peritoneal contamination, and location of injury. The complication rate for patients requiring a colostomy was 57% versus 42% for the primary repair group, P=0.01. When adjusted for injury severity and hypotension, the presence of a colostomy was not associated with a significant increase in the complication rate (OR 1.7, 95% CI: 0.9-3.25). Independent predictors for the development of intra-abdominal abscess were hypotension on admission (OR 2.4, 95% CI: 1.1-5.8) and penetrating abdominal trauma index (PATI) score >25 (OR 4.2, 95% CI: 2.0-8.9). The complication rate for colostomy takedown was 17%. CONCLUSION: Penetrating colon injury carries a high rate of infectious morbidity. The development of infectious complications is related to the injury severity and haemodynamic status of the patient, not the type of operation performed. PMID- 12531377 TI - Ulnar procedures for post-traumatic disorders of the distal radioulnar joint. AB - Post-traumatic abnormality of the distal radioulnar joint (DRUJ) still presents a therapeutic challenge to orthopaedic surgeons. The most common difficulty is a failure to diagnose these injuries early, resulting in chronic post-traumatic disorders of the DRUJ. The main aim of therapy is to avoid adverse sequelae. This is of particular importance in malunion of the distal radius, the most common cause for post-traumatic disorders of the DRUJ. Distal radial malunion can be avoided by early appropriate treatment and the need for subsequent ulnar procedures reduced. Ulnar procedures for post-traumatic disorders of the DRUJ are intended to improve function and to decrease pain. Many methods to improve post traumatic DRUJ function have been described. This article reviews the current state of the art in dealing with post-traumatic disorders of the distal radioulnar joint and presents algorithms to help in decision making. PMID- 12531378 TI - Rock-paper-scissors. AB - A study of 20 doctors in their first or second postgraduate year highlighted deficiencies in their knowledge of the nerve supply of the hand and forearm. The children's game rock-paper-scissors (Figs. 1-3) can be used as a simple aide memoir for the nerve supply to the hand and forearm. The median nerve creates the "rock position" of the pronated fist (Fig. 1). The radial nerve extends the wrist and hand forming the "paper position" (Fig. 2a and b) and the ulnar nerve creates the "scissor position" (Fig. 3), by clawing the ring and little fingers and spreading the index and middle and adducting the thumb and flexing the interphalangeal joint. PMID- 12531379 TI - The presentation of scaphoid non-union. AB - A retrospective study of 30 patients presenting with scaphoid non-union was performed. The results showed that eight had been treated in plaster for a mean of 6 weeks and discharged from follow-up. They presented with established non union at a mean of 60.3 months after the original fracture. Three of these cases have resulted in litigation against the surgeon. A total of 27 patients underwent bone grafting and Herbert's screw fixation achieving in union in 24. The natural history of scaphoid non-union has been examined in some detail in the literature. These studies have shown that scaphoid non-union is inevitably associated with arthritic changes that become worse with time and that freedom from symptoms is not protective from these changes. Therefore, prompt diagnosis and treatment of scaphoid non-union is vital, whether symptomatic or not, to prevent osteoarthritis developing in the future. Inadequate follow-up of treated scaphoid fractures will result in delayed diagnosis and treatment of non-union with a potentially impaired outcome and even litigation against the surgeon. It is the authors' advice that all scaphoid fractures treated in a plaster cast are reassessed clinically and radiologically 6 months post-injury. PMID- 12531380 TI - Late complications following clavicular fractures and their operative management. AB - We describe our experience of the operative treatment of sequelae from high energy injuries of the clavicle. Surgery was performed on 17 patients who often had more than one indication. This included the following: painful non-union (nine patients), neurogenic thoracic outlet syndrome (TOS), hypertrophic non union three patients, hypertrophic malunion five patients, bony spike two patients), vascular TOS (two patients) and brachial plexus neuroma (one patient). At clinical review, functional outcome was assessed by using a Constant score, a visual analogue pain score, return to work status and by patient subjective satisfaction rating. We performed 11 decompressions with callus excision, three simple neurolysis, eight fixations for non-union, one osteotomy and three excisions of bone spikes. Two patients required a further decompression procedure; one patient developed deep sepsis requiring sequestrectomy and two patients required removal of metalwork. At mean follow-up of 49 months the average Constant score was 64 (range 27-93). The mean pain score was 4. Ten patients returned to work at an average of 8 months. Five patients were completely asymptomatic, eight reported occasional paraesthesiae and three had significant cold intolerance. Patient satisfaction scores were 14% very satisfied, 50% satisfied, 14% unsatisfied and 22% very unsatisfied. High-energy clavicle injuries can cause significant morbidity due to non-union and TOS. Refractory symptoms can be improved by operative intervention with satisfactory functional outcome. However, the majority of patients experience some degree of residual symptoms. PMID- 12531381 TI - Extensor pollicis longus entrapment after Smith's fracture. PMID- 12531382 TI - Osteomyelitis following a haematoma block. PMID- 12531385 TI - Functions of human DNA polymerases eta, kappa and iota suggested by their properties, including fidelity with undamaged DNA templates. AB - Human DNA polymerases eta, kappa and iota are template-dependent, Y-family DNA polymerases that have been implicated in translesion DNA synthesis (TLS) in human cells. Here, we briefly review evidence that these exonuclease-deficient polymerases copy undamaged DNA with very low fidelity and unusual error specificity. Based on the base substitution specificity and other biochemical properties of DNA polymerases eta and iota, we consider the possibility that they participate in specialized DNA transactions that repair damaged DNA and/or generate mutations in the variable regions of immunoglobulin genes. PMID- 12531386 TI - Lethality in PARP-1/Ku80 double mutant mice reveals physiological synergy during early embryogenesis. AB - Ku is an abundant heterodimeric nuclear protein, consisting of 70- and 86-kDa tightly associated subunits that comprise the DNA binding component of DNA dependent protein kinase. Poly(ADP-ribose) polymerase-1 (PARP-1) is a 113-kDa protein that catalyzes the synthesis of poly(ADP-ribose) on target proteins. Both Ku and PARP-1 recognize and bind to DNA ends. Ku functions in the non-homologous end joining (NHEJ) repair pathway whereas PARP-1 functions in the single strand break repair and base excision repair (BER) pathways. Recent studies have revealed that PARP-1 and Ku80 interact in vitro. To determine whether the association of PARP-1 and Ku80 has any physiological significance or synergistic function in vivo, mice lacking both PARP-1 and Ku80 were generated. The resulting offspring died during embryonic development displaying abnormalities around the gastrulation stage. In addition, PARP-1-/-/Ku80-/- cultured blastocysts had an increased level of apoptosis. These data suggest that the functions of both Ku80 and PARP-1 are essential for normal embryogenesis and that a loss of genomic integrity leading to cell death through apoptosis is likely the cause of the embryonic lethality observed in these mice. PMID- 12531387 TI - Interactions among the Escherichia coli mutT, mutM, and mutY damage prevention pathways. AB - We have investigated in detail the interactions between the Escherichia coli mutT, mutM, and mutY error-prevention systems. Jointly, these systems protect the cell against the effects of the oxidative stress product, 8-oxoguanine (8-oxoG), a base analog with ambiguous base-pairing properties, pairing with either A or C during DNA synthesis. mutT mutator strains display a specific increase in A.T- >C.G transversions, while mutM and mutY mutator strains show specific G.C-->T.A increases. To study in more detail the in vivo processing of the various mutational intermediates leading to A.T-->C.G and G.C-->T.A transversions, we analyzed defined A.T-->C.G and G.C-->T.A events in strains containing all possible combinations of these mutator alleles. We report three major findings. First, we do not find evidence that the mutT allele significantly increases G.C- >T.A transversions in either mut(+), mutM, mutY or mutMmutY backgrounds. We interpret this result to indicate that incorporation of 8-oxodGTP opposite template C may not be frequent relative to incorporation opposite template A. Second, we show that mutT-induced A.T-->C.G transversions are significantly reduced in strains carrying mutY and mutMmutY deficiencies suggesting that 8 oxoG, when present in DNA, preferentially mispairs with dATP. Third, the mutY and mutMmutY deficiencies also decrease A.T-->C.G transversions in the mutT(+) background, suggesting that, even in the presence of functional MutT protein, A.T ->C.G transversions may still result from 8-oxodGTP misincorporation. PMID- 12531388 TI - Action of human apurinic endonuclease (Ape1) on C1'-oxidized deoxyribose damage in DNA. AB - Oxidative damage to DNA includes diverse lesions in the sugar-phosphate backbone. The chemical "nuclease" bis(1,10-phenanthroline)copper complex [(OP)(2)Cu] is believed to generate a mixture of direct oxidative strand breaks and C1'-oxidized abasic sites (2-deoxyribonolactone; dL). We found that, under our conditions, the lesions produced by (OP)(2)Cu (50 microM) in synthetic duplex DNA were predominantly dL, accompanied by approximately 30% direct strand breaks with 3' phosphates. For enzymatic studies, (OP)(2)Cu was used to introduce damage with limited sequence-selectivity, while photolysis of a site-specific 2'-deoxyuridine 1'-t-butyl ketone generated dL at a defined position. The results showed that Ape1, the major human abasic endonuclease, catalyzed 5'-incision of dL sites, but acted at least 10-fold less effectively to remove the 3'-phosphates at direct strand breaks. Kinetic analysis of Ape1 incision using the site-specific dL substrate revealed the same k(cat) for dL and regular (glycosylase-generated) abasic sites, but with K(m) approximately five-fold higher for dL substrate. The efficiency of Ape1 acting on dL, and the abundance of this enzyme in vivo, indicate that dL sites in vivo would be rapidly processed by the endonuclease. The recent observation that Ape1-cleaved dL sites can covalently trap DNA polymerase beta during the abasic excision process suggests that efficient incision of dL by Ape1 may potentiate further problems in DNA repair. PMID- 12531389 TI - Identification of the human HEX1/hExo1 gene promoter and characterization of elements responsible for promoter activity. AB - HEX1/hExo1 is a Class III nuclease of the RAD2 family with 5' to 3' exonuclease and flap structure-specific endonuclease activities. HEX1/hExo1 is expressed at low levels in a wide variety of tissues, but at higher levels in fetal liver and adult bone marrow, suggesting HEX1/hExo1 is important for hematopoietic stem cell development. A putative HEX1/hExo1 promoter fragment extending from -6240 to +1600bp exhibits cell-type specific activity in transient transfection assays. This fragment directs high luciferase reporter gene expression in the hematopoietic cell line K562, chronic myelogenous leukemia cells, but low luciferase expression in the non-hematopoietic cell line HeLa, human cervical carcinoma cells. Deletion studies identified a fragment spanning -688 to +1600bp that exhibits full transcriptional activity while a slightly shorter fragment from -658 to +1600bp exhibits significantly decreased promoter activity. In vitro binding assays revealed DNA-binding activities that interact with -687 to -681bp and -665 to -658bp elements. Oligonucleotide competition and antibody disruption studies determined that the transcription factor CREB-1 recognizes the -687 to 681bp element, while transcription factors Sp1 and Sp3 recognize the -665 to 658bp element. Mutation of either the CREB-1 or Sp1/Sp3 binding sites dramatically reduces HEX1/hExo1 promoter activity and elimination of both elements abolishes promoter activity. PMID- 12531390 TI - Repair of the mutagenic DNA oxidation product, 5-formyluracil. AB - The oxidation of the thymine methyl group can generate 5-formyluracil (FoU). Template FoU residues are known to miscode, generating base substitution mutations. The repair of the FoU lesion is therefore important in minimizing mutations induced by DNA oxidation. We have studied the repair of FoU in synthetic oligonucleotides when paired with A and G. In E. coli cell extract, the repair of FoU is four orders of magnitude lower than the repair of U and is similar for both FoU:A and FoU:G base pairs. In HeLa nuclear extract, the repair of FoU:A is similarly four orders of magnitude lower than the repair of uracil, although the FoU:G lesion is repaired 10 times more efficiently than FoU:A. The FoU:G lesion is shown to be repaired by E. coli mismatch uracil DNA glycosylase (Mug), thermophile mismatch thymine DNA glycosylase (Tdg), mouse mismatch thymine DNA glycosylase (mTDG) and human methyl-CpG-binding thymine DNA glycosylase (MBD4), whereas the FoU:A lesion is repaired only by Mug and mTDG. The repair of FoU relative to the other pyrimidines examined here in human cell extract differs from the substrate preferences of the known glycosylases, suggesting that additional, and as yet unidentified glycosylases exist in human cells to repair the FoU lesion. Indeed, as observed in HeLa nuclear extract, the repair of mispaired FoU derived from misincorporation of dGMP across from template FoU could promote rather than minimize mutagenesis. The pathways by which this important lesion is repaired in human cells are as yet unexplained, and are likely to be complex. PMID- 12531391 TI - Accumulation of 8-oxoguanine in the cellular DNA and the alteration of the OGG1 expression during ischemia-reperfusion injury in the rat kidney. AB - During ischemia-reperfusion (I/R) injury in the rat kidney, apoptosis was observed in the distal tubules of the cortico-medullary region and outer medulla (OM) while severe necrosis was seen in the proximal straight tubules of the OM. The majority of these changes disappeared within 2 weeks. We examined the contents of 8-oxo-2'-deoxyguanosine (8-oxo-dG), which is a major type of oxidative damage in DNA, in the rat kidney during I/R injury, and also investigated the expression level of the OGG1 gene encoding the 8-oxoguanine DNA glycosylase. High-performance liquid chromatography with an MS/MS analysis of the nuclear DNA revealed an immediate accumulation of 8-oxo-dG in the nuclear DNA prepared from the cortex and OM of the kidney 1h after I/R, and an immunohistochemical analysis demonstrated the immediate accumulation of 8-oxo-dG in the nuclei of renal tubular cells both in the cortex and OM. A delayed increase of cytoplasmic staining with anti-8-oxo-dG was observed only in the cortico-medulla and OM, where the cytoplasmic staining in the proximal tubular cells is higher than in the distal tubular cells. The level of cytoplasmic staining representing 8-oxo-dG in mitochondrial DNA, peaked at 6h after I/R and preceded the necrosis of proximal tubular cells in the OM. An RNase protection assay showed a high level of OGG1 mRNA in the normal kidney, and the level decreased within 3h only in the OM, and increased thereafter 1-7 days of I/R both in the cortex and OM. In situ hybridization showed higher levels of OGG1 mRNA expression in the renal tubules in the OM than in the cortex of the normal kidney, which decreased rapidly within 3h of I/R. Thus, the accumulation of 8-oxo dG in the mitochondrial DNA rather than in nuclear DNA is likely to be involved in the pathogenic responses such as necrosis of renal tubular cells during I/R injury of the kidney, together with an altered level of OGG1 expression. PMID- 12531392 TI - Cross-links between Fanconi anaemia and BRCA2. AB - Fanconi anaemia (FA) is a rare cancer-prone syndrome associated with a defect in the repair of DNA cross-links. Six genes involved in FA have been previously cloned and characterised. Now the two remaining subtypes (FA-B and FA-D1) have been shown by Howlett et al. to be associated with mutations in BRCA2 and to express truncated BRCA2 proteins. Their results suggest that the six cloned FA genes are linked with BRCA2 in a common pathway. Here Steve West discusses some of the implications of these findings. PMID- 12531393 TI - Workshop on DNA repair and related DNA transactions, a conference report. PMID- 12531394 TI - A thermodynamic model of the sympathetic and parasympathetic nervous systems. AB - In light of the nonequilibrium thermodynamics by I. Prigogine, the autonomic nervous system as a whole may be viewed as a dissipative structure progressively assembled in the course of evolution, plastically and rhythmically interfaced between forebrain, internal and external environments, to regulate energy, matter and information exchanges. In the present paper, this hypothesis is further pursued to verify whether the two main divisions of the autonomic nervous system, the sympathetic and parasympathetic systems, may support different types of exchange with the external environment. Previous data from hypothalamic stimulation experiments, studies of locus coeruleus function and available data on behavioral functional organization indicate that (1) tight engagement with the external environment, (2) high level of energy mobilization and utilization and (3) information mainly related to exteroceptive sensory stimulation characterize a behavioral prevalence of sympathoadrenal activation. On the other hand, (1) disengagement from the external environment, (2) low levels of internal energy and (3) dominance of proprioceptive information characterize a behavioral prevalence of vagal tone. Behavioral matter exchanges such as feeding, drinking, micturition and defecation are equally absent at the extreme of sympathoadrenal and vagally driven behaviors. The autonomic nervous system as a whole is genetically determined, but the sympathoadrenal system has been mainly designed to organize the visceral apparatus for an action to be performed by the biological system in the external environment and to deal with the novelty of task and of the environment, while the functional role of the parasympathetic is to prepare the visceral apparatus for an action to be performed by the biological system on itself, for recovery and self-protection (homeostasis), and is reinforced by repetition of phylo- and ontogenetically determined patterns. The available clinical data further support this interpretation indicating that an increased sympathetic and a decreased vagal tone may represent a consistent risk factor for cardiovascular diseases. PMID- 12531395 TI - Hormonal influence on expression and functionality of alpha1-adrenoceptor in rat submandibular gland. AB - In this work, we characterized alpha(1)-adrenoceptor expression and functionality in rat submandibular gland. Cumulative dose-response curve of methoxamine was constructed to determine the peroxidase secretion by glands from proestrous, estrous, metestrous and diestrous rats. They were compared to those from animals untreated or treated with sex hormones, estradiol and progesterone. The sensitivity of glands to an alpha(1)-adrenoceptor agonist varied depending on hormonal state, i.e. glands from proestrous and estrous were more sensitive to the stimulatory action of methoxamine than those from metestrous, diestrous and ovariectomised animals. The efficacy of the alpha(1) agonist was enhanced in glands from ovariectomised estrogen-treated rats but it was ineffective in glands from ovariectomised progesterone-treated rats. The functional studies correlated with 3H-prazosin binding assays in which estrogen increased alpha(1)-adrenoceptor density while progesterone decreased it. The results demonstrated that alpha(1) adrenoceptor expression and functionality in rat submandibular glands are apparently under hormonal control and probably represent other examples of bidirectional interactions between neuronal and exocrine systems. PMID- 12531396 TI - Involvement of glutamate in gastrointestinal vago-vagal reflexes initiated by gastrointestinal distention in the rat. AB - Vago-vagal reflexes play an integral role in the regulation of gastrointestinal function. Although there have been a number of reports describing the effects of various stimuli on the firing rates of vagal afferent fibers and vagal motor neurons, little is known regarding the neurotransmitters that mediate the vago vagal reflexes. In the present work, we investigated the role of glutamate in the vago-vagal reflex induced by gastrointestinal distention. Using single-cell recording techniques, we determined the effects of gastric and duodenal distention on the firing rates of gut-related neurons in the dorsal vagal complex, in the absence and presence of glutamate antagonists. Kynurenic acid, a competitive glutamate receptor antagonist, injected into the dorsal vagal complex, blocked the neuronal response of neurons in the dorsal motor nucleus of the vagus and the nucleus of the solitary tract to gastrointestinal distention. Injection of glutamate into the nucleus of the solitary tract produced inhibition of dorsal motor nucleus of the vagus neurons that were also inhibited by gastric and/or duodenal distention. Thus, the distention-induced inhibition of dorsal motor nucleus of the vagus neurons may be mediated by glutamate-induced excitation of gut-related nucleus of the solitary tract neurons. To investigate the role of the various glutamate receptor subtypes in the distention-induced events, we studied the effects of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), a selective non-NMDA receptor antagonist, and DL-2-amino-5-phosphonopentanoic acid (DL-AP5), a selective NMDA receptor antagonist. CNQX injected into the dorsal vagal complex either blocked or attenuated the inhibitory response of the neurons in the dorsal motor nucleus of the vagus and nucleus of the solitary tract neurons to gastric and duodenal distention. In contrast, DL-AP5 had less effect, especially in the vago-vagal reflex elicited by gastric distention. The results suggest (1) distention activates vagal afferents in the gastrointestinal tract; (2) the central branches of the vagal afferents from the gut terminate in the nucleus of the solitary tract and release glutamate that mainly act on non-NMDA receptors; (3) glutamate activates the inhibitory neurons in the nucleus of the solitary tract that project to the dorsal motor nucleus of the vagus; and (4) the inhibitory neurotransmitter suppresses the activity of the dorsal motor nucleus of the vagus neurons. For the excitatory neuronal responses of the dorsal motor nucleus of the vagus neurons to gastrointestinal distention, the possible circuit is that the vagal afferents containing glutamate directly activate the receptors on the dendrites of the dorsal motor nucleus of the vagus. PMID- 12531397 TI - Autonomic control of protein production by the parotid gland of the sheep. AB - The role of adrenoceptors in the control of parotid salivary function has been investigated in anaesthetized sheep. The enhancement of parotid protein output that occurs when the parasympathetic and sympathetic innervations to the gland are stimulated simultaneously in bursts at a low frequency (20 Hz for 1 s at 10-s intervals) was effectively abolished by pretreatment with propranolol (> or = 1.0 mg kg(-1), i.v., P < 0.001), without a comparable reduction in the flow of saliva or in the output of sodium or potassium. Secretion of protein was similarly augmented by simultaneous stimulation of the sympathetic innervation and an intracarotid infusion of acetylcholine (0.4-0.6 microg min(-1) g gland(-1)). This effect was also abolished by pretreatment with propranolol. Pretreatment with phentolamine (>1.0 mg kg(-1), i.v.) had no effect on the output of protein that occurred during combined stimulation of the parasympathetic and sympathetic innervations but increased the flow of saliva and the output of electrolytes. Stimulation of the parasympathetic innervation to the parotid gland caused a substantial fall in vascular resistance, which was reduced by the administration of atropine (0.5 mg kg(-1)). Stimulation of the sympathetic innervation caused a substantial rise in parotid vascular resistance in atropinized sheep. This effect was greater during continuous stimulation than during intermittent stimulation and enhanced by pretreatment with propranolol. It was virtually eliminated by pretreatment with phentolamine. It is concluded that the enhancement of protein output from the ovine parotid gland, that occurs during combined stimulation of the parasympathetic and sympathetic innervations at relatively low frequencies, depends upon interaction between cholinergic muscarinic and beta-adrenergic receptors. The vasoconstriction that occurs during sympathetic stimulation alone can be accounted for by activation of alpha-adrenoceptors. PMID- 12531398 TI - Correlation of electrophysiology, shape and synaptic properties of myenteric AH neurons of the guinea pig distal colon. AB - Well-defined correlations between morphology, electrophysiological properties and the types of synaptic inputs received are established for myenteric neurons in the guinea pig ileum. However, in the distal colon, the correlations between AH electrophysiological properties, presence of fast excitatory post-synaptic potentials (EPSPs) and neuronal shape have been inadequately resolved and it is unknown whether any colon neurons receive synaptic inputs that generate sustained excitation. In this work, we have used intracellular recording, dye filling via the recording electrode, and immunohistochemistry to classify distal colon neurons. Neurons (24 of 168) had Dogiel type II morphology and 42% of these were dendritic type II neurons, compared to about 10% in the ileum. All Dogiel type II neurons had AH electrophysiological properties, including a prolonged post-spike after-hyperpolarization (AHP). None of these received fast excitatory post synaptic potentials, 11 of 22 tested exhibited sustained slow post-synaptic excitation (SSPE) in response to 1 Hz pre-synaptic stimulation and 13 of 15 tested were immunoreactive for calbindin. Neurons (127) had Dogiel type I, filamentous or other uniaxonal cell shape and S type electrophysiology. Neurons of this group had fast excitatory post-synaptic responses to stimulation of synaptic inputs, but did not exhibit a prolonged post-spike after hyperpolarization or sustained slow post-synaptic excitation. Another group of neurons (17) had both AH electrophysiological characteristics and fast excitatory post-synaptic potentials. These neurons had Dogiel type I, filamentous or other uniaxonal shapes, but none had Dogiel type II morphology and none showed sustained slow post-synaptic excitation. It is concluded that Dogiel type II neurons are all AH neurons and are probably intrinsic sensory neurons that could be involved in long-term changes in excitability in the colon. All other neurons are monoaxonal; these are motor neurons and interneurons, and most are S neurons, electrophysiologically. A small number of monoaxonal neurons display AH electrophysiology and also receive fast excitatory synaptic inputs. These include motor and interneurons, but not sensory neurons. PMID- 12531399 TI - Antagonism of glutamatergic metabotropic receptors in the NTS of awake rats does not affect the gain of the baroreflex. AB - There is evidence suggesting that metabotropic receptors may play a role in the neurotransmission of the baroreflex in the nucleus tractus solitarius (NTS) of rats. In a recent study from our laboratory, we verified that microinjection of a metabotropic receptor agonist, trans-1-amino-1,3-cyclopentanediocarboxylic acid, into the NTS of awake and anesthetized rats produced baroreflex-like responses (hypotension and bradycardia). In the present study, we evaluated the possible role of L-glutamate metabotropic receptors of the NTS in the neuromodulation of the parasympathetic component of baroreflex activation in awake rats. Bilateral microinjection (50 nl) of a metabotropic receptor antagonist (alpha-methyl-4 carboxyphenylglycine, MCPG, 100 mM) into the rostral commissural NTS produced no change in the gain of the baroreflex bradycardia. In addition, microinjection of MCPG into the NTS produced no changes in baseline mean arterial pressure (MAP) or heart rate (HR), indicating that metabotropic receptors play no tonic role in the neurotransmission of the baroreflex. The dose of MCPG used to block the metabotropic receptors was effective in reducing the bradycardic and hypotensive responses to microinjection (50 nl) of trans-1-amino-1,3 cyclopentanediocarboxylic acid (5 mM) into the NTS. The data show that metabotropic glutamate receptors play no major role in the neuromodulation of the parasympathetic component of the baroreflex at the NTS level. PMID- 12531400 TI - Quantitative immunohistochemical investigation of the intrinsic vasodilator innervation of the guinea pig lingual artery. AB - The vasculature of the guinea pig tongue is supplied by parasympathetic vasodilator nerve fibres of intrinsic origin. Here, we investigated first to what extent neuropeptides and the synthesizing enzymes of NO, CO and acetylcholine are contained and colocalized within periarterial lingual vasodilator axons of intrinsic origin. Then it was determined whether perivascular innervation by these fibre types changes with vascular diameter, in particular in comparison with the sensory substance P (SP)-positive and sympathetic noradrenergic vascular innervation. To this end, single, double and triple labelling histochemical techniques were performed on control tongues and tongues kept in short-term organotypic culture to induce degeneration of extrinsically originating nerve fibres. Cell bodies of intrinsic microganglia and their periarterial axons contained, simultaneously, NO synthase, vasoactive intestinal peptide and the acetylcholine-synthesizing enzyme choline acetyltransferase. Additionally, neuropeptide Y (NPY) was observed in a small percentage (12%) of neurons that increased to 39% after 36 h of organotypic culture. The CO synthesizing enzyme heme oxygenase-2 was detected only in perikarya but not in periarterial axons. Intrinsic vasodilator fibres were invariably present at arteries down to a luminal diameter of 150 microm, and reached 65% of section profiles of smallest arterioles, while noradrenergic and substance P-positive axons reached 80% of arteriolar profiles. These findings show that the intrinsic lingual vasodilator innervation of the guinea pig is far extending although slightly less developed than that by sensory and sympathetic axons, and differs both in this aspect and in patterns of colocalization from that reported for other organs, e.g. lung and pelvic organs. PMID- 12531401 TI - The effect of walking on regional blood flow and acetylcholine in the hippocampus in conscious rats. AB - Recent studies in our laboratory have demonstrated that stimulation of the septal complex (i.e., the medial septal nucleus and the nucleus of the diagonal band) increases extracellular acetylcholine (ACh) release and, consequently, results in an increase in regional cerebral blood flow in the hippocampus (Hpc CBF) via activation of the nicotinic ACh receptors (nAChRs) [Neurosci. Lett. 107 (1989) 135; Neurosci. Lett. 112 (1990a) 263]. The present study aimed to examine the effects of walking on Hpc CBF, measured by laser Doppler flowmetry, in conscious rats. Walking at a moderate speed (4 cm/s) on a treadmill for 30 s produced increases in Hpc CBF and mean arterial pressure (MAP), reaching 107 +/- 1% and 105 +/- 1% of the prewalking control values, respectively. Walking for 3 min produced an increase in ACh release in the extracellular space of the hippocampus. The increase in Hpc CBF during walking was attenuated by mecamylamine (abbreviated as MEC here; 2 mg/kg, i.v.), a nAChR antagonist permeable to the blood-brain barrier (BBB), but not by hexamethonium (denoted as C6 here; 20 mg/kg, i.v.), a nAChR antagonist impermeable to the BBB, while the walking-induced increase in MAP was abolished by either agent. The response of Hpc CBF and MAP were not altered by atropine (abbreviated as ATR here; 0.5 mg/kg, i.v.), a muscarinic AChR antagonist permeable to the BBB. The increase in Hpc CBF during walking was attenuated by N(omega)-nitro-L-arginine methyl ester (L-NAME, 3 and 30 mg/kg, i.v.), a nitric oxide synthase (NOS) inhibitor, and the reduced responses were reversed following the intravenous (i.v.) administration of a physiological precursor of NO, L-arginine (600 mg/kg). The results suggest that the increase in Hpc CBF during walking is independent of MAP and attributable at least to activation of the nAChRs by the cholinergic vasodilator nerves projecting to the hippocampus and to production of NO in the hippocampus. PMID- 12531402 TI - Whole body heating reduces the baroreflex response of sympathetic nerve activity during Valsalva straining. AB - The present experiment was carried out to examine the possible effects of whole body heating on the baroreflex control of muscle sympathetic nerve activity (MSNA) and heart rate (HR). Nine healthy male volunteers underwent the Valsalva maneuver during exposure to heat stress using a hot water-perfused suit. To exclude an effect from the unloading of the cardiopulmonary baroreceptors during hyperthermia, the central hypovolemia was counterbalanced by head-down tilt (HDT) at 8 degrees. The baroreflex slopes of MSNA and HR against mean arterial pressure were determined by regression analysis during the early phase II (falling blood pressure period) of the Valsalva maneuver. The core temperature increased from 36.9 +/- 0.1 degrees C at thermoneutral to 37.7 +/- 0.1 degrees C during heat exposure, whereas the left atrial diameter (LAD) decreased. Both the MSNA and HR increased (P < 0.05) during hyperthermia. The baroreflex slopes of MSNA and HR in the phase II decreased 30 +/- 8% (P < 0.05) and 24 +/- 9% (P < 0.05), respectively, during hyperthermia. The HDT increased LAD, but there was no change in HR, arterial pressure and the baroreflex slopes. These results suggest that hyperthermia reduces the baroreflex responses of MSNA and HR during a transient falling period of blood pressure. These reduced baroreflex responses may not be attributable to an unloading of the cardiopulmonary baroreceptors during heat stress. PMID- 12531403 TI - Measures of cardiovascular autonomic regulation derived from spontaneous methods and the Valsalva maneuver. AB - The response of heart rate to a given change of systolic blood pressure is a fundamental characteristic of the cardiovascular system. The assessment of baroreflex gain (BRS) as an index of baroreflex function is based on the quantification of RR interval changes related to blood pressure changes. The spontaneous sequence and cross spectral methods describe baroreflex gain derived from spontaneous fluctuations of these parameters, yielding the up sequence and down sequence BRS and the alfa index. Phase IV of the Valsalva maneuver is also used to calculate cardiac vagal baroreflex gain. In this study, we compared the two spontaneous methods and the Valsalva maneuver in assessing baroreflex gain in 56 healthy volunteers. The BRS values calculated from different methods were as follows: up sequence BRS 12 +/- 8.6 ms/mm Hg, down sequence BRS 10 +/- 6.1 ms/mm Hg, low frequency alfa index 12.1 +/- 8.2 ms/mm Hg, Valsalva BRS 9.7 +/- 7.2 ms/mm Hg. We found close relationship between baroreflex gain derived from up and down sequences (R = 0.91, p < 0.001), down sequence BRS and low frequency alfa index (R = 0.81, p < 0.001); significant correlation between up sequence BRS and low frequency alfa index (R = 0.65, p < 0.001), the Valsalva-derived BRS and down sequence BRS (R = 0.37, p = 0.043), but no correlation between the Valsalva BRS and up sequence BRS, the Valsalva BRS and low frequency alfa index. BRS values calculated by different methods decreased with increasing age. There was no influence of age on mean arterial blood pressure elevation in late phase II of the Valsalva maneuver, nor any indication that the Valsalva BRS was related to the MAP changes. We concluded that all of these methods are useful in calculating baroreflex gain, but owing to the differences in underlying physiological mechanisms, they are not necessarily in correlation with each other. PMID- 12531404 TI - Heart rate-dependent electrocardiogram abnormalities in patients with postural tachycardia syndrome. AB - We recently published data suggesting the presence of an intrinsic sinus node abnormality in a subgroup of patients with the postural tachycardia syndrome (POTS). Based on the hypothesis that more widespread abnormalities of cardiac electrophysiologic properties may be present in POTS, we undertook a study to compare cardiac conduction and repolarization at different heart rate levels in patients with POTS and healthy controls. Eleven healthy controls and fourteen patients with POTS participated in the study. Acquisition of 12-lead electrocardiogram recordings were made during supine rest and during gradual head up tilt. The heart rate of controls was titrated by isoproterenol infusion to match the heart rate of patients. Indices for cardiac conduction (PR interval, QRS duration, and R wave axis) and repolarization (QT interval, QTc interval, and T wave axis) were then compared at different heart rate levels. The PR interval decreased with increasing heart rate in controls more than in patients, resulting in a significantly longer PR interval in patients at the fastest heart rate level. The QT and QTc intervals were significantly shorter in POTS over the entire analyzed heart rate range. The T wave axis decreased with increasing heart rate in patients only. This resulted in a significantly lower T wave axis in patients at the fastest heart rate level. Our data suggest abnormalities of atrioventricular conduction and ventricular repolarization in patients with POTS. These findings may reflect intrinsic cardiac electrophysiologic abnormalities or may be secondary due to abnormalities of cardiac autonomic innervation. PMID- 12531405 TI - Spectral analyses of electroencephalography and heart rate variability during sleep in normal subjects. AB - We investigated the relationship between electroencephalogram (EEG) activity and autonomic nervous system function using spectral analyses of EEG and heart rate variability (HRV) in healthy subjects during sleep. Eleven subjects were enrolled in this study. From EEG, the spectral edge frequencies (SEFs including SEF50, SEF90 and SEF95) were calculated. From electrocardiogram (ECG), the spectral powers of low-frequency band (LF: 0.04-0.15 Hz), high-frequency band (HF: 0.15 0.4 Hz) and the ratio of LF to HF (LF/HF) were calculated. During sleep, each set of data was obtained as the average of a 5-min measurement. We found that SEFs and LF/HF or LF decreased simultaneously and periodically, suggesting simultaneous depression of EEG activity and relative sympathetic activity, and SEFs significantly correlated with LF/HF and LF in all subjects during sleep, but not with HF. The existence of a clear correlation of SEFs with LF or LF/HF may offer a simple approach to estimate the relationship between EEG activity and autonomic nervous system function during sleep. PMID- 12531406 TI - Skin sympathetic nerve function during sleep--a study with effector responses. AB - We investigated the effector-organ activities corresponding to skin sympathetic nerve activity (SSNA) during sleep in eight healthy adult volunteers. The following parameters of SSNA were recorded during night sleep: the spontaneous skin vasoconstriction and skin blood flow volume, by laser Doppler flowmetry; sweating, by the ventilated capsule method; and the galvanic skin response (GSR). Fluctuations of sweating and GSR were mainly observed on the dorsal side of the hand during night sleep. The frequency of GSR and sweat rate on the dorsal side of the hand were significantly lower during REM sleep than during NREM sleep. The frequency of spontaneous skin vasoconstriction was higher and blood flow was lower during REM sleep than during NREM sleep. These results indicate that sweating and blood flow in the skin are differentially regulated depending on the sleep stage. Our results also suggest that the sleep-regulating system is closely linked to thermoregulation, which is controlled by the sympathetic nervous system. PMID- 12531408 TI - The pathology of heart transplant biopsy specimens: revisiting the 1990 ISHLT working formulation. PMID- 12531409 TI - Clinical predictors of exercise capacity 1 year after cardiac transplantation. AB - BACKGROUND: The exercise capacity of cardiac transplant recipients is reduced compared with normal controls. However, clinical variables predictive of post transplant exercise capacity have not been well defined. The objective of the present study was to identify clinical features predictive of post-transplant exercise capacity. METHODS: Ninety-five cardiac transplant recipients underwent cardiopulmonary testing at 1 year after transplant. The exercise parameters were compared with both pre-transplant values and normal subjects. The relationships between exercise parameters and clinical characteristics were analyzed. RESULTS: Mean peak oxygen consumption (VO(2)) and exercise test duration at 1-year post transplant improved significantly from 16.4 to 19.9 ml/kg/min and 5.5 to 7.6 minutes, respectively (p < 0.001), but were significantly lower than for normal controls (peak VO(2) 34.0 ml/kg/min; exercise duration 11.2 minutes; p < 0.001). Age- and gender-adjusted VO(2) was 54% of predicted. Pre-operative body weight correlated strongly with post-transplant weight (r = 0.80, p < 0.001). Significant recipient predictors of 1-year post-transplant peak VO(2) identified by multivariate regression analysis were age, male gender, body mass index, exercise peak heart rate and duration of post-operative intensive care. Donor variables did not contribute significantly to post-transplant peak VO(2). CONCLUSIONS: Peak VO(2) improved after cardiac transplantation but remained significantly impaired compared with normal subjects. In estimating the impact of cardiac transplantation on exercise capacity the most important pre-transplant factors to consider are age, gender and height and weight (or, alternatively, body mass index). PMID- 12531410 TI - Use of extracorporeal life support as a bridge to pediatric cardiac transplantation. AB - BACKGROUND: Extracorporeal life support (ECLS) has been used for post-cardiotomy rescue, but its use as a bridge to heart transplantation (OHT) in patients with post-surgical or end-stage ventricular failure remains controversial. METHODS: Records were reviewed for patients receiving ECLS for ventricular failure from January 1991 to August 2001. Patients listed for OHT were analyzed separately. Listing for OHT requirements were improbable myocardial recovery, absence of contraindications (central nervous system damage, high pulmonary resistance, ongoing infection, etc.), and parental consent. Outcome variables included patient demographics, diagnosis, days from ECLS initiation to United Network for Organ Sharing (UNOS) listing (latency), list time, renal function, and survival to discharge. RESULTS: Of 145 patients with ventricular failure who received ECLS, 21 pediatric patients were UNOS listed. Of 124 non-listed patients, 57 (46%) survived to discharge. All but 3 survivors were separated from ECLS in /=II) by intravascular ultrasound and normal coronary angiography. A higher mean first year biopsy score may be associated with significant transplant coronary artery disease by intravascular ultrasound, but a large number of patients will be required to determine this with statistical certainty. One major complication occurred early in the experience. CONCLUSIONS: In children, intravascular ultrasound is more sensitive for detecting significant transplant coronary artery disease than coronary angiography, but may add cost, time and potential morbidity to screening protocols. Prospective, multicenter studies are needed to best utilize intravascular ultrasound in this patient population. PMID- 12531411 TI - Concomitant allorecognition of mismatched donor HLA class I- and class II-derived peptides in pediatric lung transplant recipients with bronchiolitis obliterans syndrome. AB - BACKGROUND: The authors' previous studies with 2 different adult patient populations demonstrated a correlation between indirect allorecognition of mismatched donor HLA Class I- and Class II-derived peptides and the development of bronchiolitis obliterans syndrome (BOS) after lung transplantation. The aim of this study was to determine whether a parallel allorecognition of mismatched donor HLA Class I- and Class II-derived peptides occurs after lung transplantation and to determine its correlation with the development of BOS after lung transplantation in a group of pediatric patients. METHODS: Peripheral blood mononuclear cells from 7 BOS-positive and 6 BOS-negative pediatric lung transplant recipients (age, 11.5 +/- 4.4 years) were cultured in the presence of synthetic peptides corresponding to the alpha-chain hypervariable regions of a mismatched donor HLA Class I molecule and the beta-chain hypervariable region of a mismatched donor HLA-DR molecule. The frequencies of HLA Class I and Class II alloreactive T cells were determined using limiting dilution analysis. RESULTS: A significant increase (p = 0.025) in HLA Class I-alloreactive T cells was observed in BOS-positive patients (7.1 x 10(-5) +/- 4.3 x 10(-5)) compared with BOS negative patients (2.1 x 10(-5) +/- 1.8 x 10(-6)). In addition, a significant increase (p = 0.033) in HLA Class II-alloreactive T cells also was observed in BOS-positive patients (9.6 x 10(-5) +/- 7.9 x 10(-5)) compared with BOS-negative patients (1.3 x 10(-5) +/- 2.1 x 10(-6)). CONCLUSIONS: This study indicates that a parallel CD4+ T-cell alloreactivity to both donor HLA Class I and Class II molecules may play a role in the pathogenesis of BOS both in adult and pediatric lung transplant recipients. PMID- 12531413 TI - Time-related changes in pulmonary function after conversion to tacrolimus in bronchiolitis obliterans syndrome. AB - BACKGROUND: Bronchiolitis obliterans syndrome (BOS) is a leading cause of morbidity and mortality after lung and heart-lung transplantation. Present treatment is directed at the augmentation of pharmacologic immunosuppression. METHODS: This study examines the effect of substituting cyclosporine with tacrolimus on the forced expiratory volume in 1 second (FEV(1)) and on the forced expiratory flow between 25% and 75% of vital capacity (FEF(25%-75%)) in 32 patients who developed BOS. The proportional rates of decline of FEV(1) and FEF(25%-75%) before and after treatment with tacrolimus were calculated. The actuarial survival of responders and non-responders to tacrolimus was compared. Pre-operative and post-operative factors were investigated to determine any difference between the 2 groups. RESULTS: There were significant reductions in the rates of decline of FEV(1) and FEF(25%-75%) when the rates in the 3 months before conversion to tacrolimus were compared with subsequent rates at 0 to 3 months, 3 to 6 months, 6 to 9 months and 9 to 12 months after conversion. The rates of decline of FEV(1) and FEF(25%-75%) in the 3 months before conversion were 0.11 liters/month and 0.13 liters/s per month, respectively. This compares with the rates of decline for FEV(1) and FEF(25%-75%) for the 3 months after conversion to tacrolimus of 0.04 liters/month (p = 0.023) and 0.04 liters/s per month (p = 0.022), respectively. The actuarial survival at 1 year from the time of conversion to tacrolimus for the responder sub-group and the non-responder sub group were 89.2% and 75%, respectively, and at 4 years after conversion were 61.3% and 56.3%, respectively (p = 0.92). CONCLUSIONS: Tacrolimus rescue therapy is effective at stabilizing lung function in patients with BOS, and this effect is apparent up to 12 months after conversion from cyclosporine. PMID- 12531414 TI - Humoral rejection in cardiac transplantation: risk factors, hemodynamic consequences and relationship to transplant coronary artery disease. AB - BACKGROUND: Acute cellular rejection is the mechanism of most immune-related injury in cardiac transplant recipients. However, antibody-mediated humoral rejection (HR) has also been implicated as an important clinical entity following orthotopic heart transplantation. Humoral rejection has been reported to play a role in graft dysfunction in the early post-transplant period, and to be a risk factor for the development of transplant coronary artery disease. Some involved in transplantation pathology doubt the existence of clinically significant humoral rejection in cardiac allografts. Those who recognize its existence disagree on its possible role in graft dysfunction or graft coronary artery disease. In this study, we report clinical features of patients with the pathologic diagnosis of HR at our institution since July 1997, when we began systematic surveillance for humoral rejection. METHODS: We reviewed medical records of patients with the pathologic diagnosis of HR without concurrent cellular rejection between July 1997 and January 2001. Diagnosis was based on routine histology ("swollen cells" distending capillaries, interstitial edema and hemorrhage) and immunofluorescence (capillary deposition of immunoglobulin and complement with HLA-DR positivity), or immunoperoxidase staining of paraffin embedded tissue (numerous CD68-positive macrophages and fewer swollen endothelial cells distending capillaries). RESULTS: A total of 44 patients (4 to 74 years old) showed evidence of HR without concurrent cellular rejection at autopsy or on one or more biopsies. Although females comprised only 26% of our transplant population, 23 patients (52%) with HR were female. A positive peri-operative flow cytometry T-cell crossmatch was observed in 32% of HR patients compared with 12% of controls (p = 0.02). Hemodynamic compromise consisting of shock, hypotension, decreased cardiac output/index and/or a rise in capillary wedge or pulmonary artery pressure was observed in 47% of patients at the time of diagnosis of HR. Six patients (5 females) died (14% mortality) with evidence of HR at or just before autopsy, 6 days to 16 months after transplantation. The incidence of transplant coronary artery disease was 10% greater at 1 year, and 36% greater at 5 years, in patients with HR when compared with non-HR patients. CONCLUSIONS: Humoral rejection was associated with acute hemodynamic compromise in 47% of patients, and was the direct cause of death in 6 patients (13%). Humoral rejection is a clinicopathologic entity with a high incidence in women and is associated with acute hemodynamic compromise, accelerated transplant coronary artery disease and death. PMID- 12531415 TI - The effect of beta-adrenergic blockers on the prognostic value of peak exercise oxygen uptake in patients with heart failure. AB - OBJECTIVES: Our aim was to determine the effect of beta-adrenergic blockade on the prognostic value of peak oxygen consumption testing in patients with heart failure. BACKGROUND: Peak oxygen consumption has been shown to be a useful prognostic tool in patients with heart failure. However, studies demonstrating the utility of peak oxygen consumption were conducted before beta-blocker therapy became widespread. Thus, our objective was to determine the effect of beta blockers on the prognostic value of peak oxygen consumption in patients with heart failure. METHODS: Actuarial, anthropomorphic, hemodynamic and exercise ventilatory data were collected from 369 patients with heart failure. Death and orthotopic heart transplants were the events tracked. Patients were divided into those taking beta-blockers and those not taking them. Event-free survival days were calculated. RESULTS: One hundred ninety-nine patients on beta-blockers and 170 not on beta-blockers were studied. There were 40 orthotopic heart transplants and 82 deaths during follow-up. Peak oxygen consumption (milliliters per kilogram per minute) trended toward being an independent predictor of event-free survival (p = 0.055). In patients on and not on beta-blockers, a peak oxygen consumption of >14 ml/kg.min was associated with a 1-year event rate of approximately half of that associated with a peak oxygen consumption 80 years, n = 92); group 2 (<80 years, n = 276). RESULTS: There were significant differences between the two groups with regard to the sex ratio, the American Society of Anesthesiologists (ASA) classification, the emergency presentation, and the curative operation rate. The operative mortality rate was 8% in group 1, 4% in group 2 (P = 0.26). The overall 5-year survival rate was 35% in group 1, 53% in group 2 (P = 0.0004). In patients operated on for cure, the cancer-specific 5-year survival rate was 50% in group 1, 59% in group 2 (P = 0.08). CONCLUSIONS: The prognosis for rectal cancer in patients over 80 years is not significantly different from that of younger patients. Surgery should not be restricted on the basis of age. PMID- 12531447 TI - Hyperparathyroidism associated with thyroid pathology. AB - BACKGROUND: The pathological association between thyroid and parathyroid gland disease is here discussed. The multiphase analyzer has revealed a new type of subclinical primary hyperparathyroidism (HPP) and the role of surgery in these cases is not clear. METHODS: This is a prospective study of all cases of thyroid disease in association with parathyroid disease treated surgically in our Institute from July 1999 to June 2001. RESULTS: Of the 221 thyroidectomies carried out, 29 patients had an elevated preoperative serum level of parathyroid hormone (PTH). An ultrasonography examination was performed on all patients and a preoperative scanning with 99Tc-MIBI on 11 of 29 patients. We examined intraoperatively 19 cases of HPP (14 parathyroid adenoma, 5 hyperplasia). In 10 cases we observed a normal size of the parathyroid gland and we did not perform a parathyroidectomy. CONCLUSIONS: All patients with elevated serum parathyroid hormone and serum calcium levels before thyroidectomy should be considered candidates also for surgery to the parathyroid glands. The pathological association between thyroid and parathyroid gland diseases is not rare. We must conduct an accurate neck exploration in all these cases. PMID- 12531448 TI - Comparison of repair techniques for major incisional hernias. AB - BACKGROUND: Multiple techniques exist for repair of incisional hernias. Laparoscopic ventral hernia repairs as well as sliding myofascial rectus flap ventral hernia repairs (the separation of parts technique) have received recent attention for low recurrence and low complication rates. These procedures are diametrically opposed solutions to the same clinical problem. METHODS: Two surgeons of different surgical specialties will jointly compare and contrast these two ends of the spectrum regarding repair of ventral hernias. RESULTS: An algorithm for patient selection is presented. CONCLUSIONS: No one technique is the "best" procedure for all patients. Knowledge of the wide variety of surgical options will be of benefit to all patients with incisional hernias. PMID- 12531449 TI - Partial splenectomy using a coupled saline-radiofrequency hemostatic device. AB - BACKGROUND: Partial splenectomy is indicated for benign tumors and cysts of the spleen, as well as, operative management of splenic trauma limited to one pole of the spleen. Despite improved technique, bleeding from the cut surface of the spleen still remains an obstacle. METHODS: We describe our technique for partial splenectomy using a new device based on coupling saline with radiofrequency energy to achieve hemostasis while dividing the splenic parenchyma. RESULTS: Use of this technique has led to blood loss of less than 50 cc, while achieving splenic preservation. PMID- 12531450 TI - Hemipelvectomy for severe decubitus ulcers in patients with previous spinal cord injury. AB - BACKGROUND: Patients with spinal cord injury (SCI) frequently have pressure ulcers. Surgery is sometimes needed to close them. In rare cases, hemipelvectomy is warranted for extremely severe complications. METHODS: We conducted a retrospective study using national Department of Veterans Affairs (DVA) computer data sets to identify clinical features of SCI patients who underwent hemipelvectomy for life-threatening septic complications of decubitus ulcers. RESULTS: Among the approximately 4 million patients receiving care in the DVA system, more than 40,000 patients were treated on an inpatient basis for SCI during the search period (fiscal years 1989 to 1998). They represent approximately 20% of the total national patient pool. There were 56 patients who supposedly had undergone hemipelvectomy. Chart review eliminated cases that did not meet our inclusion criteria, resulting in 8 evaluable cases. All had complete SCI due to trauma and later developed severe pressure sores with pelvic osteomyelitis or life-threatening soft tissue infection. CONCLUSIONS: This series is the largest reported to date. The surgery involved significant blood loss (mean 2.6 L). Reoperations and complications were common. The mortality rate was 25%, but the survivors were all markedly improved by the surgery. Some of the complications appeared to be related more to the SCI than to the pelvic sepsis or surgery, suggesting that meticulous perioperative care may be valuable in reducing the complication rate in SCI patients undergoing this radical operation for very severe sequelae of pressure ulcers. PMID- 12531451 TI - Negotiation skills for physicians. AB - As stakeholders vie for increasingly limited resources in health care, physicians would be well advised to hone their skills of negotiation. Negotiation is defined as a strategy to resolve a divergence of interests, be they real or perceived, where common interests also exist. Negotiation requires effective communication of goals, needs, and wants. The "basic needs" model of negotiation is best suited to the current health care environment. In this model, negotiator must to be able to identify their needs in the negotiation, establish their best alternative to a negotiated agreement, and identify their strategies and tactics for the negotiation. PMID- 12531452 TI - The effects of an additive small amount of a low residual diet against total parenteral nutrition-induced gut mucosal barrier. AB - BACKGROUND: Total parenteral nutrition (TPN) negatively influences the gut mucosal barrier. It has been suggested that enteral nutrition is effective against the harmful influence. METHODS: Forty-eight male Donryu rats underwent placement of a central venous catheter and tube gastrostomy. They were divided into six groups, receiving isocaloric nutrients in various proportions of PN and a low residual diet (LRD) for 7 days. RESULTS: Intestinal permeability, villous height and crypt depth, and number of secretory IgA-positive cells in the villus were measured. Intestinal permeability was significantly reduced in rats receiving an LRD corresponding to more than 15% of total caloric intake. Gut morphological structure was maintained in rats receiving an LRD corresponding to more than 10%. A higher number of IgA-positive cells was observed in rats receiving an LRD corresponding to more than 15%. CONCLUSIONS: A small amount of LRD could prevent decreases in gut mucosal integrity. There was a stepwise defense mechanism in the gut mucosal barrier. PMID- 12531453 TI - Von Recklinghausen disease and gastrointestinal stromal tumors. AB - BACKGROUND: We discuss how the notion of mesenchymal or connective tumors of the digestive tract has passed to the concept of stromal tumors and its links with von Recklinghausen disease, and make special reference to the progress in medical treatment. METHODS: We have studied one of our own cases and did a review of the literature. RESULTS: For isolated primary lesions, good results are obtained with surgery. For metastatic forms, dramatic improvements have been recently described with STI 51, an inhibitor of tyrosium-kinase. CONCLUSIONS: Gastrointestinal stromal tumors are now well understood with immunochemistry analysis. Their links with von Recklinghausen disease deserve attention and are not probably a random event. Primary isolated tumors are best treated by surgery; metastatic or recurrent forms have recently shown dramatic improvement with STI 571. PMID- 12531454 TI - Inadvertent transmural India ink tattooing simulating intestinal infarction. PMID- 12531455 TI - Relationship between musculoskeletal symptoms and blood markers of oxidative stress in patients with chronic fatigue syndrome. AB - In 21 patients with chronic fatigue syndrome (CFS) versus 20 normal subjects, we investigated the oxidant/antioxidant balance and its correlation with muscle symptoms. Patients versus controls showed significantly: lower Lag Phase and Vitamin E (Vit E) concentrations in plasma and low-density lipoproteins (LDL), higher LDL thiobarbituric acid reactive substances (TBARS), higher fatigue and lower muscle pain thresholds to electrical stimulation. A significant direct linear correlation was found between fatigue and TBARS, thresholds and Lag Phase, thresholds and Vit E in plasma and LDL. A significant inverse linear correlation was found between fatigue and Lag Phase, fatigue and Vit E, thresholds and TBARS. Increased oxidative stress and decreased antioxidant defenses are related to the extent of symptomatology in CFS, suggesting that antioxidant supplementation might relieve muscle symptoms in the syndrome. PMID- 12531456 TI - Modulation of cytokine mRNA expression by brain-derived neurotrophic factor and nerve growth factor in human immune cells. AB - Brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) exert various effects on immune cells. Here we studied, whether they influence the cytokine expression pattern in peripheral blood mononuclear cells (PBMCs) or antigen specific T-cells. In PBMCs BDNF and NGF had interindividually variable effects on T helper cell type (Th)1- and Th2-cytokines. However, there was a high correlation between the modulating properties of these neurotrophins (r=0.97) concerning the expression of interleukin (IL) 4, transforming growth factor-beta and tumour necrosis factor-alpha mRNA at a concentration of 100 ng/ml. In myelin basic protein-specific T-cell lines BDNF and NGF increased interferon -gamma mRNA to a moderate extent, but not IL4. No major effects were detected at the cytokine protein level. In conclusion, our results suggest a partial effect of neurotrophins on immune cells, which may be modified by other signals. PMID- 12531457 TI - Dissociative increase of oligodendrocyte progenitor cells between young and aged rats after transient cerebral ischemia. AB - To investigate the effect of aging on the oligodendrocyte progenitor cells (OPCs) after cerebral ischemia, neuron-glia antigen 2 (NG2) chondroitin sulfate proteoglycan was examined at 1, 3 and 7 days after 90 min of transient middle cerebral artery occlusion in young and aged brains. The number of NG2 positive cells increased in the ischemic penumbra at 3 and 7 days after reperfusion, while those decreased in the ischemic core. At 7 days, the number of NG2 positive cells was significantly greater in the young than the aged brains, and the processes of NG2 positive cells enlarged and were highly branched in the young than the aged brains. These results suggest that the young brain showed a higher potential of proliferation and process branching of OPCs than the aged brains. PMID- 12531458 TI - The role of oxidative stress in paraquat-induced neurotoxicity in rats: protection by non peptidyl superoxide dismutase mimetic. AB - Herbicides, including paraquat, may produce neurodegenerative effect when given both peripherally and into the brain though the pathophysiological mechanism is still unknown. Microinfusion of paraquat into the Substantia Nigra (50 microg) produced increased motor activity, jumping and circling opposite to the injection site, associated with ECoG desynchronization, high voltage epileptogenic spikes, and with neuropathological effects. These effects were accompanied by increase of malondialdehyde (MDA) levels in the Substantia Nigra, suggesting that paraquat was able to induce oxidative stress when injected directly into the rat brain. Pre-treatment of rats with M40401, a non peptidyl superoxide dismutase (SOD) mimetic given directly into the Substantia Nigra or i.p. prevented both behavioural, electrocorticogram and neuropathological effects and MDA elevation. Taken together, these results demonstrate that paraquat produces brain damage via abnormal formation of oxygen free radicals and that this effect may be counteracted by novel SOD mimetics. PMID- 12531459 TI - Neuroprotective effects of anticonvulsants in rat hippocampal slice cultures exposed to oxygen/glucose deprivation. AB - Some anticonvulsants show neuroprotective effects, and may be of use in reducing neuronal death resulting from stroke or traumatic brain injury. Here I report that a broad range of anticonvulsants protect cells in hippocampal slice cultures from death induced by oxygen/glucose deprivation (OGD). Hippocampal slice cultures were submitted to 1 h OGD and the resulting cell death was quantified 24 h later using a novel automated fluorescent scanning method. The classical anticonvulsants phenobarbital, phenytoin, ethosuximide, chlordiazepoxide and midazolam all significantly and dose-dependently reduced cell death induced by OGD. The newer anticonvulsants carbamazepine, felbamate, lamotrigine, tiagabine, and oxcarbazepine also had significant neuroprotective effects, but gabapentin, valproic acid (10 mM), levetiracetam and retigabine were not neuroprotective at a concentration up to 300 microM. In conclusion, several classical and newer anticonvulsants have neuroprotective properties in an in vitro model that simulates cerebral ischemia. PMID- 12531460 TI - Spatial asymmetries of auditory event-synthesis in humans. AB - We used the mismatch negativity event-related potential to examine how spatial location and feature variation affect the capacity of the auditory system to automatically respond to pairs of rapid (180 ms apart) acoustic changes within a single tone. When a tone first deviated from a standard tone in source location and then in its duration, we found independent responses to both deviations for right but not left field stimuli. In contrast, when the first deviation was in pitch and the second in duration, only the first deviation elicited a response, regardless of presentation side. These results suggest that information from either side of space is asymmetrically processed even in a free-field, and that the extent of the temporal window of integration is not a fixed property of the auditory system. PMID- 12531461 TI - Neurotensin expressing neurons developed earlier than vasoactive intestinal polypeptide and vasopressin expressing neurons in the human suprachiasmatic nucleus. AB - Development of neurotensin (NT), vasoactive intestinal polypeptide (VIP), vasopressin (AVP) and neuropeptide-Y (NPY) expressing neurons was investigated in the human fetal suprachiasmatic nucleus of nine subjects ranging from 20-40 weeks of gestation using immunocytochemistry and morphometry. Results obtained showed that NT expressing neurons developed earlier than VIP, AVP and NPY expressing neurons. Consistent with results obtained from animal studies, we also found VIP expressing neurons were born earlier than AVP expressing neurons. Whether the NT expressing neurons play a role in generating circadian rhythms in the early life of humans needs to be further investigated. PMID- 12531462 TI - Hyperhomocysteinemia as a new risk factor for brain shrinkage in patients with alcoholism. AB - Chronic alcohol consumption can induce brain atrophy, whereby the exact mechanism of brain damage in alcoholics remains unknown. There is evidence that chronic alcoholism is associated with hyperhomocysteinemia. Homocysteine is an excitatory amino acid which markedly enhances the vulnerability of neuronal cells to excitotoxic and oxidative injury in vitro and in vivo. The present volumetric magnetic resonance imaging study included 52 chronic alcoholics and 30 non drinking healthy controls. Patients were active drinkers and had an established diagnosis of alcohol dependence. We investigated the influence of different variables on the hippocampal volume of patients suffering from chronic alcoholism. We observed that pathological raised levels of plasma homocysteine showed the most significant correlation to hippocampal volume reduction (P<0.001, multiple regression analysis). Raised plasma levels of homocysteine are associated with hippocampal (brain) atrophy in alcoholism. PMID- 12531463 TI - Functional specialization within the anterior medial prefrontal cortex: a functional magnetic resonance imaging study with human subjects. AB - This study investigated the functional neuroanatomy of the anterior medial prefrontal cortex (aMPFC). Previous studies have shown that the aMPFC is involved in evaluative judgment and self-referential processes. Specifically, different sections of the aMPFC are differentially influenced by attention demanding processes. Whereas the dorsal section is supposed to be involved in self referential processes, the ventral section is assumed to be attenuated during attention demanding processes. The present study investigates the involvement of the dorsal and ventral aMPFC in evaluative judgment by using functional magnetic resonance imaging with spin-echo echo-planar-imaging. Processes involved in evaluative judgment are attention-demanding, self-referential and activate regions in the dorsal and ventral section of the aMPFC. Attention demanding tasks do not necessarily lead to an attenuation of the ventral section of the aMPFC, a region mainly involved in emotional and affective processing. PMID- 12531464 TI - Mechanisms underlying the cardiovascular responses to spinal dopamine receptor stimulation by apomorphine in anesthetized rats. AB - The present study investigated the mechanisms by which intrathecal (i.t.) apomorphine affects mean aortic pressure and heart rate in anesthetized rats. In saline-pretreated rats, upper thoracic (T2-T4) i.t. administration of apomorphine (48 microg/rat) induced immediate and significant hypotension and bradycardia. These responses were unaffected by intravenous (i.v.) methylatropine (1 mg/kg) or bilateral vagotomy, while they were prevented by i.t. lidocaine (25 microl at 1%) or i.v. hexamethonium (30 mg/kg). However, i.v. atenolol (1.5 mg/kg) suppressed the apomorphine-induced bradycardia without affecting the hypotension in either intact or bivagotomized rats. Bilateral adrenalectomy had no effect upon both maximal hypotensive and bradycardic responses to apomorphine (48 microg/rat at the T9-T10 level). These results suggest that hypotensive and bradycardic responses to i.t. apomorphine are due to an action in the spinal cord, presumably on sympathetic preganglionic neurons. These responses are dissociated and seem to result from withdrawal of sympathetic outflow to the vasculature and to the heart, respectively. PMID- 12531465 TI - Functional magnetic resonance imaging mirrors recovery of visual perception after repetitive tachistoscopic stimulation in patients with partial cortical blindness. AB - We investigated three patients with partial cortical blindness after brain injury by means of functional magnetic resonance imaging (fMRI) before and after the application of a daily visual stimulation-therapy over a period of 6 months. Before therapy, fMRI data showed a severely reduced blood-oxygen-level-dependent (BOLD) signal in primary visual cortex when compared to healthy volunteers. Following several months of rehabilitative therapy a neuropsychological improvement of visual functions was accompanied by an increase in BOLD signal of residual perilesional regions whereas fMRI data of the control group remained unchanged. A high capacity of functional recovery and synaptic plasticity of surviving perilesional neuronal structures of primary visual cortex followed by an increased input into post-connected visual areas can be discussed as a basis for the reoccurrence of visual functions. PMID- 12531466 TI - Metastin-like immunoreactivity in the rat medulla oblongata and spinal cord. AB - Metastin, the product of metastasis suppressor gene KiSS-1, is proposed to be the natural ligand for the G-protein-coupled receptor GPR54, known also as AXOR12. This immunohistochemical study, using a rabbit polyclonal antiserum against the human metastin fragment (45-54)-NH(2), showed that in rats metastin-like immunoreactivity (MTS-LI) was present in neurons of the nucleus of the solitary tract and caudoventrolateral reticular nucleus, and in cell processes of the spinal trigeminal tract and lateral reticular nucleus. MTS-LI was confined mainly to neurons and fibers at or caudal to the area postrema. In the spinal cord, MTS LI cell processes formed a dense plexus in superficial layers I and II of the dorsal horn. The pattern of distribution of MTS-LI in the medulla and spinal cord suggests that this novel peptide may participate in autonomic and sensory neural signaling. PMID- 12531467 TI - Differential cerebellar activation related to perceived pain intensity during noxious thermal stimulation in humans: a functional magnetic resonance imaging study. AB - Little is known about the cerebellar involvement in pain processing in spite of the fact that the cerebellum probably plays a crucial role in pain-related behavior. Using functional magnetic resonance imaging we examined the differential cerebellar activation in 18 healthy subjects in relation to their perceived pain-intensity of noxious and non-noxious thermal stimuli. In contrast to non-noxious (40 degrees C) stimuli, noxious (48.5 degrees C) stimuli revealed activation in the deep cerebellar nuclei, anterior vermis and bilaterally in the cerebellar hemispheric lobule VI. With the same noxious stimulus (48.5 degrees C) there was differential cerebellar activation depending on the perceived pain intensity: high pain intensity ratings were associated with activation in ipsilateral hemispheric lobule III-VI, deep cerebellar nuclei and in the anterior vermis (lobule III). This differential cerebellar activation pattern probably reflects not only somatosensory processing but also perceived pain intensity that may be important for cerebellar modulation of nociceptive circuits. PMID- 12531468 TI - Changed vesicular GABA transporter immunoreactivity in the gerbil hippocampus following spontaneous seizure and vigabatrin administration. AB - To identify the roles of vesicular gamma-aminobutyric acid (GABA) transporter (VGAT) in epileptogenesis and the recovery mechanisms in spontaneous seizure, we conducted a chronological and comparative analysis of VGAT expression. VGAT immunoreactivity was stronger in the seizure resistant group than that in the pre seizure group of seizure sensitive (SS) gerbils. In 3 h postictal group, the density of VGAT immunoreactivity was significantly increased in the hippocampus, as compared to pre-seizure group. In 24 h postictal group, VGAT immunodensity had recovered to its pre-seizure level. In addition, VGAT immunoreactivity in the hippocampus was also increased by vigabatrin (GVG) administration. These results suggest that decreased VGAT expression in the SS gerbil hippocampus may affect epileptogenesis in this animal, and that the subsequent alteration in its expression induced by seizure and the administration of GVG may reflect a modulation of GABA release to alleviate seizure activity. PMID- 12531469 TI - Effects of 3,4-methylenedioxymethamphetamine on locomotor activity and extracellular dopamine in the nucleus accumbens of Fischer 344 and Lewis rats. AB - Previous studies have shown that Fischer 344 (F344) and Lewis (LEW) rats may differ with respect to their behavioural and neurochemical responses to several drugs of abuse, including amphetamines. Herein, we have examined whether such strain differences extend to a ring-substituted amphetamine, namely 3,4 methylenedioxymethamphetamine (MDMA, ecstasy), a recreationally-used drug endowed with euphoric, but also long-term neurotoxic effects. Beside strain differences in baseline locomotor activity (F344>LEW), it was found that the subcutaneous administration of 10 mg/kg, but not 5 mg/kg, MDMA increased locomotor activity in F344 rats only. On the other hand, such a treatment increased to similar extents extracellular dopamine (DA) levels in the nucleus accumbens of F344 and LEW rats, thus suggesting that genetic differences in MDMA locomotor effects are not accounted for by accumbal DA release. PMID- 12531470 TI - A subset of nociceptin/orphanin FQ receptor-expressing neurons in the anterior hypothalamic area, as revealed in mice with lacZ reporter gene. AB - Nociceptin/orphanin FQ (N/OFQ) is an endogenous peptide agonist for the opioid receptor homolog, N/OFQ receptor, and serves for the central control of autonomic functions. Morphological details including the cell types that may account for such N/OFQ functions, however, remain unclear. By using X-gal histochemistry for the detection of receptor-expressing cells at both light and electron microscopic levels, we examined the hypothalamus from the receptor-deficient mice bearing a lacZ insertional mutation in the N/OFQ receptor gene. The N/OFQ receptor reflected by lacZ expression was seen at high levels in the anterior hypothalamic area. With electron microscopy, lacZ expression was observed in a subset of neurons showing large cell size and indented nucleus. PMID- 12531471 TI - Two novel genes are differentially expressed during early germination of the male gametophyte of Nicotiana tabacum. AB - Two cDNAs were cloned from tobacco pollen by differential display of mRNAs from dry and germinated pollen. Expression analysis revealed that transcript levels of the two corresponding genes were high during pollen maturation, but differed significantly during early pollen germination. One of the cDNAs, designated Arpl1;1, codes for a new member of the auxin-repressed/dormancy-associated protein multigene family. High Arpl1;1 transcript levels correlated with pollen maturation. In germinating pollen, transcripts rapidly declined to minimal levels. The second cDNA, designated Mads1;11, codes for a novel transcription factor exhibiting a novel structure for a putative MADS box protein from plants. Transient expression of a chimeric gene encoding the N-terminal part of Mads1;11 fused to the green fluorescent protein (GFP) in onion epidermal cells revealed that the protein was localized to the nucleus, consistent with its presumed function as a transcriptional regulator. Mads1;11 mRNA abundance was high in mature pollen and, in contrast to Arpl1;1, remained high during 6 h of in vitro pollen germination. The data indicate differential regulation of mRNA concentrations during pollen germination and suggest a function of Mads1;11 protein in regulating gene expression during early pollen tube growth. PMID- 12531472 TI - Isolation and characterization of four cell wall purple acid phosphatase genes from tobacco cells. AB - Four full-length cDNAs were isolated from a cDNA library prepared from tobacco cultured cells and designated NtPAP4, NtPAP12, NtPAP19 and NtPAP21, which could correspond to purple acid phosphatase (PAP). Levels of both NtPAP12 and NtPAP21 mRNA in the protoplasts immediately increased after the protoplasts were transferred to a medium for cell wall regeneration, and the accumulation of the mRNA was correlated with cell wall regeneration for 3 h. It is likely that the NtPAP12 and NtPAP21 gene products are wall-bound PAPs at the early stage of regenerating walls in tobacco protoplasts. PMID- 12531473 TI - Human transformer 2beta and SRp55 interact with a calcitonin-specific splice enhancer. AB - The calcitonin/calcitonin gene-related peptide (CGRP) pre-mRNA is alternatively processed in a tissue-specific manner leading to the production of calcitonin mRNA in thyroid C cells and CGRP mRNA in neurons. Sequences in the human calcitonin-specific fourth exon function as an exonic splice enhancer (ESE) which is required for incorporation of exon 4 into calcitonin mRNA. Deletion of these sequences from the rat calcitonin/CGRP gene was reported to have no effect on calcitonin splicing. We demonstrate that sequences in the rat calcitonin/CGRP fourth exon act as an ESE. In addition, we observed that three proteins in HeLa nuclear extract, of apparent molecular weights of 40, 55 and 85 kDa, specifically interact with the exon 4 ESE. The 40-kDa protein is human transformer 2beta (hTra2beta), a homolog of the Drosophila splice regulator transformer 2. hTra2beta is required for calcitonin splicing in vitro, one of the first biological functions identified for hTra2beta. The 55-kDa protein is SRp55, a member of the SR family of phosphoproteins. Binding of SRp55 to an ESE required for calcitonin mRNA splicing suggests that the different levels of SRp55 present in different cell types may regulate calcitonin/CGRP alternative splicing. PMID- 12531474 TI - Binding of a candidate splice regulator to a calcitonin-specific splice enhancer regulates calcitonin/CGRP pre-mRNA splicing. AB - The calcitonin/calcitonin gene-related peptide (CGRP) pre-mRNA is alternatively processed in a tissue-specific manner leading to the production of calcitonin mRNA in thyroid C cells and CGRP mRNA in neurons. A candidate calcitonin/CGRP splice regulator (CSR) isolated from rat brain was shown to inhibit calcitonin specific splicing in vitro. CSR specifically binds to two regions in the calcitonin-specific exon 4 RNA previously demonstrated to function as a bipartate exonic splice enhancer (ESE). The two regions, A and B element, are necessary for inclusion of exon 4 into calcitonin mRNA. A novel RNA footprinting method based on the UV cross-linking assay was used to define the site of interaction between CSR and B element RNA. Base changes at the CSR binding site prevented CSR binding to B element RNA and CSR was unable to inhibit in vitro splicing of pre-mRNAs containing the mutated CSR binding site. When expressed in cells that normally produce predominantly CGRP mRNA, a calcitonin/CGRP gene containing the mutated CSR binding site expressed predominantly calcitonin mRNA. These observations demonstrate that CSR binding to the calcitonin-specific ESE regulates calcitonin/CGRP pre-mRNA splicing. PMID- 12531475 TI - The rat histone H1d gene has intragenic activating sequences that are absent from the testis-specific variant H1t. AB - In some cases core histone genes in the mouse depend on intragenic sequence elements for high level expression [Gene 176 (1996) 1]. Here we report that the highly expressed gene for rat linker histone H1d also contains an intragenic activating region (IAR). Using transient transfection assays in mouse fibroblast NIH3T3 cells, we showed that rat H1d contains a downstream region (+21 to +116) that imparts a two- to threefold up-regulation of fused reporters. This region also activated expression when moved to the promoter region, though the effect was dependent on its distance from other promoter elements. The IAR contains sequence homologies to the core alpha and Omega elements identified as functional protein binding sites within the mouse H3.2 coding region activating sequence (CRAS). A pair of Omega elements (+32 and +66) accounts for the activating effect of the H1d intragenic region as shown by targeted mutations as well as stepwise deletions. The H1d and H3.2 Omega sequences bound similar and perhaps identical proteins by gel shift analysis. The H1d alpha-like sequence at +56 overlaps the translational start codon and was therefore not mutated. Like the mouse H3.2 alpha element, it bound transcription factor YY1 in gel shift assays. H1t, the gene for the testis-specific linker histone, did not demonstrate an IAR. While H1t has a similar alpha sequence and did bind YY1, it lacks the Omega homologies of H1d. Sequence comparison shows that the YY1/alpha site as well as the adjacent Omega site are likely present in genes for other standard H1 variants, but that the +32 Omega site in the 5' untranslated region (UTR) of H1d is unique. We conclude that the +32 and +66 Omega sequences of the rat H1d gene contribute significantly to its high-level expression. PMID- 12531476 TI - Identification and characterization of a novel cancer/testis antigen gene CAGE-1. AB - Serological analysis of cDNA expression library (SEREX) was employed to identify cancer-associated genes. By screening cDNA expression libraries with sera of patients with lung cancers, we identified a total of 49 genes that specifically reacted with the sera of patients with lung cancers. Among these, we characterized a novel gene with expression pattern similar to that of cancer/testis antigens. Its open reading frame is 1920 bp in size and encodes for putative protein composed of 639 amino acids. Southern blot analysis reveals that this gene exists as single copy. In vitro transcription/translation and Western blot analysis confirm that this gene encodes a protein of 73 kDa in size. The comparison of cDNA and genomic sequences reveals that it is composed of 11 exons and 10 introns. This gene displays testis-specific expression among normal tissues, and wide expression among various cancer tissues and cancer cell lines. A study using GFP fusion construct reveals mainly nuclear localization of CAGE-1 protein. The expression of this clone is relatively higher in cancer tissues compared with their surrounding non-cancerous tissues. This suggests that overexpression of CAGE-1 may be associated with the progression of tumor. Because of its association with cancer, this gene was named cancer-associated gene-1 (CAGE-1). Given the fact that several cancer/testis antigens reportedly induce cytolytic T lymphocyte (CTL) reactions, it is reasonable that this gene will be a valuable target for cancer immunotherapy. The exact functional role of CAGE-1 in tumorigenesis remains to be seen. PMID- 12531477 TI - Freeze-induced expression of a novel gene, fr47, in the liver of the freeze tolerant wood frog, Rana sylvatica. AB - The ability to endure the freezing of body fluids is well developed as an adaptation for winter survival in several species of woodland frogs. Recently, the mechanisms supporting natural freeze tolerance have been shown to include the expression of novel genes. One such novel gene, fr47, codes for a 390-amino acid protein present in the livers of freeze-tolerant anurans (Rana sylvatica, Pseudacris crucifer, Hyla versicolor) but not in freeze-intolerant species (Rana pipiens, Scaphiopus couchii). Regulatory influences on gene and protein expression were investigated using R. sylvatica. Northern blot analysis showed that transcript levels were increased following 24 h of freezing (5.1-fold), 24 h of anoxia exposure (6.4-fold), or the loss of 20% of total body water (2.7-fold). Immunoblotting with anti-FR47 antibody indicated that protein levels increased during freezing and thawing, but decreased somewhat during anoxia or dehydration exposure, although rebounding during recovery. These results suggest that (i) FR47 function is important for freeze survival, and (ii) that control at the protein level may be exerted posttranscriptionally. Finally, assessment of putative signal transduction pathways regulating fr47 gene expression, via in vitro incubations of liver slices, indicated the involvement of a protein kinase C-mediated pathway. PMID- 12531478 TI - Cloning and characterization of the promoter region of human spata2 (spermatogenesis-associated protein 2) gene. AB - In this paper, we report the genomic organization of the human spata2 (spermatogenesis-associated protein 2) gene and the characterization of its promoter region. The 5'-flanking region lacks canonical TATA-box but contains a CAAT-box and potential binding sites for a variety of transcription factors such as Sp1, NF-1 and CREB. The results indicated that (1) the cis-acting elements necessary for a basal transcription of spata2 are contained within the -81/+159 proximal sequence of the promoter and (2) the strongest promoter activity was exhibited by the -403/+159 region that contains a CREB site. PMID- 12531479 TI - Isolation and functional analysis of the 5'-flanking region of carrot top1beta gene coding for the beta isoform of DNA topoisomerase I. AB - We have isolated and functionally characterized the promoter region of the top1beta gene encoding carrot (Daucus carota) DNA topoisomerase Ibeta. The major transcription start site was mapped by primer extension analysis 164 nt upstream the ATG translation start codon. Sequence analysis of the 5'-upstream region of the gene revealed the presence of a canonical TATA-like box at position -35 bp and several cis-acting sequences, including a (CT)n element in the leader region of the gene, a myb-related motif and the Dof element NtBBF-1, which correlate with the inducible expression pattern of this gene. Functional reporter analysis of the top1beta 5'-flanking region was performed in both carrot and Arabidopsis thaliana transfected protoplasts. The region at -719 to +161 was sufficient to confer high expression level in both species. The transient expression assay in protoplasts induced to stop dividing confirmed that the promoter, whose activity is low in quiescent cells, is activated when protoplasts are induced to proliferate by exogenous application of growth factors. PMID- 12531480 TI - An alternatively spliced variant of cathepsin E in human gastric adenocarcinoma cells. AB - Splice variant and authentic mRNAs for procathepsin E were measured at a ratio of 5:1 and 1:2 in Kato 3 and AGS cells, two human gastric adenocarcinoma cell lines. As a result of the precise splicing of the 3'-end of exon 6 to the 5'-end of exon 8, the variant lacked the 142 bp of exon 7 which encodes the second of the Asp residues that operate the catalytic mechanism of aspartic proteinases. PMID- 12531481 TI - Cloning and characterization of the SIL promoter. AB - The SIL gene undergoes a site-specific rearrangement with the SCL gene in 25% of patients with T cell acute lymphoblastic leukemia (ALL). The functional result of this rearrangement is that the SIL regulatory elements aberrantly drive expression of the SCL gene. We have cloned and sequenced the human SIL promoter, cloned a murine homolog, found the sequence to be highly conserved, and defined a minimal promoter region. Both the cloned murine and human sequences were found to be highly active in either human or murine cells. SCL mRNA, driven by a cloned SIL promoter, could be downregulated by DMSO in stably transfected F4-6 murine erythroleukemia cells. The SIL promoter was found to be partially unmethylated in proliferating tissues, in which it is highly expressed, and more highly methylated in post-mitotic tissues, in which SIL is not expressed. The isolation of the SIL promoter provides an important tool for the study of both the SIL gene expression as well as the role of the SIL promoter in leukemogenesis. PMID- 12531482 TI - Cloning, expression and characterization of a functional cDNA clone encoding geranylgeranyl diphosphate synthase of Hevea brasiliensis. AB - Geranylgeranyl diphosphate (GGPP) synthase catalyzes the condensation of isopentenyl diphosphate (IPP) with allylic diphosphates to give (all-E)-GGPP. GGPP is one of the key precursors in the biosynthesis of biologically significant isoprenoid compounds. In order to examine possible participation of the GGPP synthase in the enzymatic prenyl chain elongation in natural rubber biosynthesis, we cloned, overexpressed and characterized the cDNA clone encoding GGPP synthase from cDNA libraries of leaf and latex of Hevea brasiliensis. The amino acid sequence of the clone contains all conserved regions of trans-prenyl chain elongating enzymes. This cDNA was expressed in Escherichia coli cells as Trx-His tagged fusion protein, which showed a distinct GGPP synthase activity. The apparent K(m) values for isopentenyl-, farnesyl-, geranyl- and dimethylallyl diphosphates of the GGPP synthase purified with Ni(2+)-affinity column were 24.1, 6.8, 2.3, and 11.5 microM, respectively. The enzyme shows optimum activity at approximately 40 degrees C and pH 8.5. The mRNA expression of the GGPP synthase was detected in all tissues examined, showing higher in flower and leaf than petiole and latex, where a large quantity of natural rubber is produced. On the other hand, expression levels of the Hevea farnesyl diphosphate synthase were significant in latex as well as in flower. PMID- 12531483 TI - Complete DNA sequence of the atp operon of the sodium-dependent F1Fo ATP synthase from Ilyobacter tartaricus and identification of the encoded subunits. AB - The atp operon of Ilyobacter tartaricus, strain DSM 2382, was completely sequenced using conventional and inverse polymerase chain reaction (i-PCR) techniques. It contains nine open reading frames that were attributed to eight structural genes of the F(1)F(o) ATP synthase and the atpI gene, which is not part of the enzyme complex. The initiation codons of all atp genes, except that of atpB coding for the a subunit, were identified by the corresponding N-terminal amino acid sequence. The hydrophobic a subunit was identified by MALDI mass spectrometry. The atp genes of I. tartaricus are arranged in one operon with the sequence atpIBEFHAGDC comprising 6,992 base pairs with a GC content of 38.1%. The F(1)F(o) ATP synthase of I. tartaricus has a calculated molecular mass of 510 kDa and includes 4,810 amino acids. The gene sequences and products reveal significant identities to atp genes of other Na(+)-translocating F(1)F(o) ATP synthases, especially in the F(o) subunits a and c which are directly involved in ion translocation. PMID- 12531485 TI - A periaxonal net in the zebrafish central nervous system. AB - We produced a monoclonal antibody, named A20, which specifically recognizes a 35 kDa protein and stains myelinated axons in zebrafish brain. The A20 antigen is located at the outside of the myelin layer of large axons, and comprises a fine meshwork composed of thin unit fibers about 1-2 microm in length and about 100 200 nm in thickness. The unit fibers form pentagonal and hexagonal structures, which further polymerize into an envelope structure on the axons. The A20 monoclonal antibody did not stain neuronal cell bodies nor synapses. Instead, the distribution of the A20 antigen was along axons, practically coincident with the distribution of myelin basic protein. The monoclonal antibody stained only axons in the central nervous system (CNS), and not the extracellular matrix surrounding Schwann cells. These results suggest that this antigenic meshwork (which we call the periaxonal net) is synthesized by oligodendrocytes. During the development of the zebrafish brain, the periaxonal net appeared after the formation of myelin on the axons. The periaxonal net developed first at the brain stem, then gradually appeared at the caudal end of the spinal cord. The thickness of the periaxonal net around the Mauthner axon changed during development. Although the thickness of the Mauthner axon continues to grow throughout life, the thickness of periaxonal net stopped growing at 6 months after fertilization. PMID- 12531486 TI - Central nuclei mediating estrogen-induced changes in autonomic tone and baroreceptor reflex in male rats. AB - The current investigation examines the significance of estrogen in central cardiovascular regulatory nuclei in modulating autonomic tone and baroreceptor reflex function. Experiments were done in anaesthetized male Sprague-Dawley rats. Changes in autonomic tone were assessed by monitoring vagal and renal efferent nerve activities before and following bilateral injection of estrogen into select central autonomic nuclei. In the first study, selective blockade of neurotransmission through the central nucleus of the amygdala (CNA), lateral hypothalamic area (LHA) and ventral posteromedial thalamic nucleus (VPM) using the local anaesthetic lidocaine was done to determine which nuclei were involved in mediating the autonomic changes observed following bilateral injections of estrogen into the insular cortex (IC). In the second study, the role of the parabrachial nucleus (PBN) in mediating the autonomic changes observed following bilateral estrogen injections into the CNA, LHA, VPM and IC was determined by blocking neurotransmission through the PBN using lidocaine.Injections of estrogen into the IC produced a significant increase in renal sympathetic nerve activity (RSNA; from 10+/-2 to 24+/-4 microV/sec; p<0.05). This estrogen-induced increase in RSNA was significantly attenuated when lidocaine was pre-injected into the LHA, CNA or PBN (55+/-6, 33+/-4 and 91+/-7% decrease respectively; p<0.05) but not when injected into the VPM (16+/-6% decrease; p>0.05). Injection of estrogen into the CNA resulted in a significant decrease in RSNA (48+/-5%; p<0.05) whereas estrogen injection into the LHA resulted in a significant increase (28+/-4%; p<0.05) in RSNA. Pre-injection of lidocaine into the PBN resulted in complete blockade of the autonomic changes observed following estrogen injection into the CNA but did not affect the changes observed following estrogen injection into the LHA. These results suggest that estrogen acting in forebrain and midbrain cardiovascular nuclei activated efferent pathways which synapse in the LHA, CNA and/or PBN prior to projecting to autonomic preganglionic nuclei to affect autonomic tone. These nuclei may therefore provide an added level of processing and/or integration of the autonomic response(s) following activation by local or systemic estrogen. PMID- 12531487 TI - Behavioral training increases local astrocytic metabolic activity but does not alter outcome of mild transient ischemia. AB - Functional neurological outcome after transient ischemia might be improved by timely therapeutic intervention. To determine if restorative behavioral therapy influences damage, improves task learning, or alters astrocyte metabolic activity after ischemia, rats (food-restricted to 85% of free-feeding weight) were (a) first trained to respond on one of two levers under a fixed-ratio 20 schedule of food presentation (FR20), then (b) subjected to sham manipulation of carotid arteries or 10 min ischemia by four-vessel occlusion, followed by (c) 4 days of operant testing or inactivity, (d) then all rats were tested under a FR20 lever reversal task for 4 weeks, and (e) 3 days after the last behavioral session astrocyte metabolism was assayed by local uptake of [2-14C]acetate. Mild loss of hippocampal neurons occurred in ischemic rats with or without training after ischemia. Glial fibrillary acidic protein-positive astrocytes were present in similar numbers throughout brains of sham control and ischemic rats. Mild ischemia did not impair learning, and no changes in FR20 reversal learning were detected in sham vs. ischemic rats. Net [14C]acetate uptake was unaffected by ischemia but [14C]acetate uptake increased 15-24% (P<0.05; n=12-15/group) in specific structures (caudate, primary motor and sensorimotor cortex, CA1 hippocampus, subcortical white matter) in the pooled groups of rats that had 4 days FR20 testing vs. inactivity before reversal learning. 'Behavioral therapy' (operant testing on the 4 days immediately following either sham manipulation or ischemia) did not alter ischemic outcome, but was associated with higher acetate utilization in regions involved in motor activities. PMID- 12531488 TI - Protective effect of nilvadipine against glutamate neurotoxicity in purified retinal ganglion cells. AB - We determined the effect of nilvadipine, a dihydropyridine-type calcium channel blocker, in preventing glutamate neurotoxicity in purified retinal ganglion cells (RGCs). RGCs were purified from dissociated rat retinal cells (postnatal days 6 8), using a modified two-step panning method, and cultured in serum-free medium containing neurotrophic factors and forskolin. RGC survival after exposure to glutamate (25 microM) with nilvadipine or other calcium channel blockers was measured by calcein-acetoxymethyl ester staining after 3 days in culture. Changes in the level of intracellular Ca(2+) ([Ca(2+)](i)) were measured with fura-2 fluorescence. Induction of apoptosis was evaluated using the TDT-dUTP terminal nick-end labeling technique. The neurotoxic effects of low doses of glutamate were blocked by a specific alpha-amino-3-dihydro-5-methylisoxazole-4-propionate kainate receptor antagonist, 6,7-dinitroquinoxaline-2,3-dione (20 microM). Simultaneous application of nilvadipine (1-100 nM) with glutamate protected against glutamate neurotoxicity in a dose-dependent manner. Calcium-imaging experiments showed that the glutamate-evoked [Ca(2+)](i) increase was significantly blocked by nilvadipine (P<0.001), but not nifedipine and diltiazem, in about 50% of RGCs. In addition, the application of nilvadipine significantly reduced glutamate-induced apoptosis (P<0.001). These findings suggest that nilvadipine may partly inhibit glutamate-induced apoptotic cell death by blocking calcium influx via voltage-dependent calcium channels in purified RGCs. PMID- 12531490 TI - Effects of transcranial magnetic stimulation on muscle activation patterns and joint kinematics within a two-joint motor synergy. AB - We studied the effects of transcranial magnetic stimulation (TMS) on muscle activation patterns and joint kinematics during oscillatory movements of a joint within a two-joint (elbow-wrist) synergy. Healthy young adults (N=8) performed cyclic movements in one of the joints while the other joint was or was not involved into a postural task. TMS was applied unexpectedly at different phases of the movement cycle. TMS-induced motor evoked potentials (MEPs) showed a modulation within the movement cycle, partly related to changes in the background muscle activity. After normalization by the background activity, MEPs were higher in conditions involving maintenance of a posture by one of the joints. MEPs in a flexor-extensor muscle pair could show peaks at the same phase within the movement cycle while background EMG peaks were out of phase. TMS induced a phase shift in the ongoing movement, and the phase shift changed over the next cycles. It stabilized by the fourth cycle and could show both phase advances and phase delays, the latter dominated. The findings are interpreted as supporting two hypotheses: (1) TMS can activate cortical neurons interposed in a transcortical long-latency loop leading to pre-programmed reactions to perturbations; and (2) apparent effects of TMS on the timing of an ongoing cyclic movement are defined by an interaction between its effects on a central 'clock' and those induced by the peripheral mechanical response. PMID- 12531489 TI - Paradoxical [correction of parodoxical] effect of orexin A: hypophagia induced by hyperthermia. AB - This experiment tested the effect of the sympathetic and thermogenic activation induced by orexin A on eating behavior. The food intake, firing rate (FR) of the sympathetic nerves to interscapular brown adipose tissue (IBAT), IBAT and abdominal temperatures (T(IBAT) and T(ab)), and heart rate (HR) were monitored in 24 h-fasting male Sprague-Dawley rats for 15 h after food presentation. Orexin A (1.5 nmol) was injected into the lateral cerebral ventricle 6 h before food presentation while FR, T(IBAT) and T(ab), and HR were also monitored. The same variables were controlled in rats receiving orexin A contemporaneously to food presentation. Two other groups of control animals were tested with the same procedure, however orexin A was substituted by saline. The results showed that food intake was significantly lower in the group receiving orexin A 6 h before food presentation in comparison to all the other groups. FR, T(IBAT) and T(ab), and HR were significantly higher in the rats receiving orexin A with respect to rats receiving saline. These findings demonstrate that orexin A, so-called for its orexigen action, can also induce hypophagia. On the other hand, orexin A always induces an activation of the thermogenesis. These results suggest a revision of the role played by orexin A in the control of food intake, assigning to this peptide a primary role in the thermoregulation. The possibility that orexin A can induce hypophagia is well demonstrated by this experiment, so that the scientific community should use a different name for this peptide. An appropriate name could be 'hyperthermine' A. PMID- 12531491 TI - EEG and ERP profiles in the high alcohol preferring (HAP) and low alcohol preferring (LAP) mice: relationship to ethanol preference. AB - Neurophysiological measures, such as decreased P300 amplitude and altered EEG alpha activity, have been associated with increased alcoholism risk. The purpose of the present study was to extend the assessment of the neurophysiological indices associated with alcohol consumption to a recently developed mouse model of high ethanol consumption, the first replicate line of high alcohol preferring (HAP-1) and low alcohol preferring (LAP-1) mice. Male HAP-1, LAP-1, and HS mice from the Institute for Behavioral Genetics at the University of Colorado Health Science Center (i.e., HS/Ibg mice) were implanted with cortical electrodes. EEG activity, and event related potentials (ERPs) were then examined. Following electrophysiological assessment, ethanol preference was assessed to examine the relationship between neurophysiological indices and ethanol consumption. EEG analyses revealed that HAPs and HS/Ibgs had greater peak frequency in the 2-4-Hz band and lower peak frequency in the 6-8- and 1-50-Hz bands of the cortical EEG compared to LAPs. Compared to HAPs, LAPs and HS/Ibgs had decreased peak EEG frequency in the 8-16-Hz band. Decreased parietal cortical power from 8 to 50 Hz was associated with high initial ethanol preference in HAP mice. In regards to ERPs, P1 amplitude was greater in HAPs compared to both LAPs and HS/Ibgs and the P3 latency in LAPs was decreased compared to both HAPs and HS/Ibgs. As expected, HAPs consumed more ethanol and had higher ethanol preference than LAPs and HS/Ibgs. There were no significant differences in ethanol intake or preference between HS/Ibgs and LAPs. These data indicate that selective breeding of the HAP and LAP lines has resulted in the divergence of EEG and ERP phenotypes. The differences observed suggest that increased cortical P1 amplitude and altered cortical EEG activity in the 8-50-Hz frequency range may be neurophysiological 'risk factors' associated with high ethanol consumption in mice. Decreased P3 latency in LAPs compared to HAPs and HS/Ibgs mice may be a 'protective factor'. PMID- 12531492 TI - Characterization of the peroxisome proliferator activated receptor coactivator 1 alpha (PGC 1alpha) expression in the murine brain. AB - The peroxisome proliferator activated receptor coactivator 1 alpha (PGC-1alpha) is a nuclear transcriptional coactivator that is expressed in brown adipose tissue, brain, heart and kidney as well as cold-exposed skeletal muscle. In liver, white and brown adipose tissue, PGC-1alpha expression is regulated in a manner suggesting a role in energy homeostasis. To characterize PGC-1alpha expression in the rodent brain and to determine brain PGC-1alpha regulation, we used in situ hybridization histochemistry in C57Bl/6J mice and Sprague-Dawley rats. We found that PGC-1alpha is widely expressed in brain areas, including in the olfactory bulb, cerebral cortex, the diagonal band of Broca, the medial septal nucleus, reticular thalamic nucleus, the striatum and globus pallidus, the hippocampus, the substantia nigra, the mesencephalic nucleus of the trigeminal nerve, the cochlear nucleus and the superior olivary complex. In contrast, PGC 1alpha expression was absent in the hypothalamus. To evaluate PGC-1alpha expression under different physiologic states in these various brain areas, we examined expression with fasting, leptin treatment and cold exposure (4 h at 4 degrees C) and found no change, nor was expression changed in the brain of the leptin-deficient ob/ob mice and the hyperleptinemic UCP-DTA mice. Hence, PGC 1alpha is widely expressed in the rodent brain, but is not regulated by states of caloric deficiency, leptin, obesity or cold exposure. Its functional role in the brain requires further study. PMID- 12531493 TI - Head position modifies cerebrovascular response to orthostatic stress. AB - Previous experiments have shown that the vestibular system participates in cardiovascular control. However, the effects of vestibular activation on cerebrovascular regulation are not known. Therefore, the present experiment tested the hypothesis that specific vestibular activations may be beneficial to cerebral circulation during simulated orthostatic stress. Middle cerebral artery blood flow velocity (CBV; Doppler ultrasound) was measured to examine the effects of head-down neck flexion (HDNF) compared to head-down neck extension (HDNE) with and without lower body negative pressure (LBNP; -40 mmHg) (n=9). The change in CBV (DeltaCBV) during HDNF and HDNE were not different during baseline conditions, however, during LBNP, DeltaCBV was greater in HDNE compared to HDNF ( 5.5+/-3.2 cm/s, -11+/-4.6%) vs. (-0.7+/-1.0 cm/s, -1.9+/-1.9%), respectively (P<0.05). Concomitantly, the change in cerebrovascular resistance (DeltaCVR) between rest and LBNP was also greater during HDNE (0.48+/-0.08 mmHg/cm per s, 42.8+/-10.8%) compared with HDNF (0.26+/-0.05 mmHg/cm per s, 22+/-4.1%) (P<0.05). P(ET)CO(2) was greater in HDNE (45+/-2 mmHg) compared to HDNF (42+/-2 mmHg; P<0.05) during LBNP. These results suggest that the vestibular system may affect cerebrovascular tone during simulated postural stress by either constriction or dilation, depending on the vestibular stimulus. PMID- 12531494 TI - Mechanisms involved in the antinociception caused by compound MV8612 isolated from Mandevilla velutina in mice. AB - The pregnane compound MV8612 isolated from the rhizome of the plant Mandevilla velutina administered by intraperitoneal (i.p.), intrathecal (i.t.) or by intracerebroventricular (i.c.v.) routes caused graded and complete inhibition of the thermal hyperalgesia caused by i.t. injection of bradykinin (BK) in mice with mean ID(50) values of 7.8 micromol/kg, 33.6 and 4.6 nmol/site, respectively. Compound MV8612 (i.p.) also inhibited both the neurogenic and inflammatory pain responses to formalin with mean ID(50) values of 5.6 and 10.6 micromol/kg, respectively. Given i.t., MV8612 produced significant inhibition of both phases of the formalin-induced licking (inhibition of 34+/-5 and 36+/-4%, respectively). Given by i.c.v. route MV8612 inhibited both phases of formalin-induced pain (32+/ 6 and 63+/-5%) with mean ID(50) of 8.4 nmol/site against the late phase. MV8612, given by i.p., i.c.v. or i.t. routes, also inhibited capsaicin-induced pain (51+/ 4, 25+/-8 and 39+/-6%, respectively). The i.t. injection of potassium (K(+)) channel blockers, apamin and charybdotoxin given 15 min before, markedly prevented the antinociception of MV8612 against both phases of formalin-induced nociception. In contrast, tetraethylammonium (TEA) or glibenclamide had no effect. The i.c.v. treatment with pertussis toxin resulted in a significant inhibition of both MV8612- and morphine-induced antinociception against both phases of formalin-induced pain. Taken together these results confirm and also extend our previous data by demonstrating that the greater part of the antinociception caused by MV8612 seems to be associated with its ability to interfere with BK action. Finally, both the low and high conductance calcium (Ca(2+))-activated K(+) channels and the activation of G(i/o) pertussis sensitive G-proteins take part in the mechanism by which compound MV8612 produces antinociception. PMID- 12531495 TI - Impact of the endothelin-A receptor antagonist BQ 610 on microcirculation in global cerebral ischemia and reperfusion. AB - The role of endogenous endothelin-1 in mediating microcirculatory disturbances after global cerebral ischemia was investigated in Mongolian gerbils. The pial microcirculation was studied by intravital fluorescent microscopy before, during, and up to 3 h after occlusion of both carotid arteries for 15 min. Pretreatment was achieved with the peptidergic selective endothelin-A (ET-A) receptor antagonist BQ 610. The neurological outcome was assessed daily for up to 4 days. The antagonist attenuated postischemic leukocyte-endothelium interactions in postcapillary venules, in particular the number of rolling leukocytes was found to be reduced (13.0+/-9.4 x 100 microm(-1) min(-1) in the control vs. 2.0+/-2.5 in the experimental group, P<0.05). The local microvascular perfusion, measured by the arterio-venous transit time, was improved during reperfusion by BQ 610 (1.3+/-0.5 s in the control vs. 0.7+/-0.2 s in the experimental group, P<0.05). The neurological deficit was significantly reduced in animals treated with the ET A antagonist (P<0.05). The inhibition of the postischemic inflammatory reaction and the reversal of the delayed hypoperfusion may account for the improved neurological outcome. These observations suggest that application of endothelin-A antagonists may be a useful approach to interfere with derangements in cerebral ischemia/reperfusion. PMID- 12531496 TI - A chloride channel blocker reduces acetylcholine uptake into synaptic vesicles at the frog neuromuscular junction. AB - A key mechanism for loading acetylcholine (ACh(+)) into synaptic vesicles uses energy to transport H(+) into the vesicle interior and then exchanges H(+) for ACh(+). This mechanism requires anions to follow the H(+) into the vesicles to prevent the building up of an overwhelming electrical gradient across the vesicle membrane. Frog nerve-muscle preparations were treated with hypertonic solution in which sodium gluconate was the major constituent, which substantially increases the sizes of the quanta by increasing their ACh(+) content. The Cl(-) channel blocker 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) antagonized the increases in quantal size, so it seems likely that Cl(-) follows H(+) to prevent the buildup of a potential gradient across the vesicular membrane. PMID- 12531497 TI - Rapid dot sputum and serum assay in pulmonary tuberculosis. AB - A rapid direct sputum (Sp.) and/or antibody assay, based on immunoblotting and enzyme immunoassay is described. The test can detect mycobacterial antigens or antibodies in clinical specimens from pulmonary tuberculosis (TB) patients. In this study, 87 sputa, 87 sera and 40 paired sputa and sera were utilized from smear-positive and smear-negative, culture-positive patients; 59 sputa, 37 sera and 22 paired sputa and sera from nontuberculosis respiratory disease patients and 68 sera from healthy controls. The antigen detection in sputum by dot assay has 86.1% sensitivity on active tuberculosis patients, 92.9% specificity, 91.6% positive predictive value (PPV), 88.2% negative predictive value (NPV) and 10.3% error. The antibody assay has 83.6% sensitivity, 95.4% specificity, 94.4% positive predictive value, 85.6% negative predictive value and 11% error. The test performed on paired sputum and serum (Sr.) samples has a sensitivity of 93.3%, which rose to 96.1% on smear-positive and culture-positive patients, but the specificity decreased to 83% in sputum, whereas in serum it was 92%. The results of the assay, combined with clinical and radiological data, could form the basis for starting an earlier course of treatment for tuberculosis. PMID- 12531498 TI - Molecular quantification of genes encoding for green-fluorescent proteins. AB - A quantitative PCR approach is presented to analyze the amount of recombinant green fluorescent protein (gfp) genes in environmental DNA samples. The quantification assay is a combination of specific PCR amplification and temperature gradient gel electrophoresis (TGGE). Gene quantification is provided by a competitively coamplified DNA standard constructed by point mutation PCR. A single base difference was introduced to achieve a suitable migration difference in TGGE between the original target DNA and the modified standard without altering the PCR amplification efficiency. This competitive PCR strategy is a highly specific and sensitive way to monitor recombinant DNA in environments like the efflux of a biotechnological plant. PMID- 12531499 TI - Application of single-strand conformation polymorphism and denaturing gradient gel electrophoresis for fla sequence typing of Campylobacter jejuni. AB - Campylobacter jejuni is a frequent cause of bacterial gastroenteritis in humans all over the world. Several molecular typing methods are used to study the epidemiology of Campylobacter spp. infections. The aim of the present study was to investigate the application of single-strand conformation polymorphism (SSCP) and denaturing gradient gel electrophoresis (DGGE) analysis as rapid primary subtyping methods for C. jejuni. A variable fragment from the 3' end of the flaA to the 3' end of the intergenic region, separating the flaA and flaB genes, was subjected to SSCP and DGGE analysis. A total of 48 clinical C. jejuni isolates, 49 C. jejuni strains isolated from poultry, 2 strains isolated from ducks and 1 strain isolated from a pheasant were assigned to 24 distinct SSCP patterns. Sequence analysis of the respective DNA fragments revealed that every different fla sequence type could be distinguished by SSCP. DGGE proved to be equally discriminatory. Both methods can be applied as primary subtyping methods, because pulsed-field gel electrophoresis (PFGE) and amplified fragment length polymorphism (AFLP) analysis further differentiated isolates belonging to the same fla sequence types. PMID- 12531500 TI - A novel approach for extraction of PCR-compatible DNA from activated sludge samples collected from different biological effluent treatment plants. AB - This paper describes a method that facilitates the extraction of PCR-compatible DNA from different activated sludge samples. The approach involves a novel preprocessing step in DNA extraction, which removes potential PCR inhibitors. The sludge was washed with different ratios of acetone and petroleum ether after pretreatment with 0.01% Tween-20 at 50 degrees C. It was observed that an initial washing step with 50 mM Tris-HCl, pH 9.0, before the detergent-solvent step, improved the quality of the extracted DNA. The extraction protocol resulted in amplifiable amounts of DNA when 10 mg of a sludge sample was used, even in the presence of phenol as a sludge contaminant. The usefulness of the extracted template was demonstrated by carrying out different PCR reactions. The random amplified polymorphic DNA (RAPD) patterns demonstrated the diversity of sludge samples. PMID- 12531501 TI - Use of gas chromatography-mass spectrometry/solid phase microextraction for the identification of MVOCs from moldy building materials. AB - Gas chromatography-mass spectrometry/solid phase microextraction (GC-MS/SPME) was applied to identify microbial volatile organic compounds (MVOCs) in water damaged, mold-infested building materials (gypsum board papers (n=2), mineral wool, and masonite) and in cultivated molds (Aspergillus penicillioides, Stachybotrys chartarum, and Chaetomium globosum). Three SPME fibers (65-microm PDMS-DVB, 75-microm Carboxen-PDMS, and 70-microm Carbowax-stableflex) designed for automated injection were used of which the latter showed best performance. A number of previously reported MVOCs were detected both in the building materials and the cultivated molds. In addition, methyl benzoate was identified both in the S. chartarum and A. penicillioides cultures and in the building materials. SPME combined with GC-MS may be a useful method for the determination of MVOCs emitted from mold-infested building materials. PMID- 12531502 TI - An improved method for the determination of NADH oxidase in the presence of NADH peroxidase in lactic acid bacteria. AB - The complexity of the coupled NADH oxidase-NADH peroxidase enzyme system in lactic acid bacteria makes it difficult to simultaneously determine the individual levels of both these enzymes spectrophotometrically. This study describes an improved assay to accurately determine low concentrations of NADH oxidase from enzyme suspensions containing NADH oxidase and NADH peroxidase. For the standardisation of the assay, pure NADH oxidase and NADH peroxidase were mixed in various proportions and the percentage recovery was estimated by both the currently available assay as well as by the improved assay reported in this study. The recovery of NADH oxidase using the currently available assay ranged from as low as -200% to as high as +102% as against 90-102% in the improved assay. The recovery percentage of NADH peroxidase ranged from 91% to 112% in both assays. The slopes of NADH oxidation by cell-free extracts of six lactic acid bacteria were also measured by both assays for the estimation of NADH oxidase and NADH peroxidase levels. The improved assay can further distinguish between NADH H(2)O oxidase and NADH-H(2)O(2) oxidase and was successfully applied to identify the type of NADH oxidase in the lactic acid bacteria tested. PMID- 12531503 TI - Evaluation of a novel method based on amplification of DNA fragments surrounding rare restriction sites (ADSRRS fingerprinting) for typing strains of vancomycin resistant Enterococcus faecium. AB - In the search for an effective DNA-typing technique for use in hospital epidemiology, the performance and convenience of a novel assay based on the fingerprinting of bacterial genomes by amplification of DNA fragments surrounding rare restriction sites (ADSRRS fingerprinting) was tested. A large number of vancomycin-resistant Enterococcus faecium (VREM) isolates from haematological ward patients of the Clinical Hospital in Gdansk were examined. We found that ADSRRS fingerprinting analysis is a rapid method that offers good discriminatory power. The method demonstrated also excellent reproducibility. The usefulness of the ADSRRS fingerprinting method for molecular typing was compared with pulsed field gel electrophoresis (PFGE) method, which is currently considered the gold standard for molecular typing of isolates recovered from patients and the environment in the course of investigation and control of nosocomial outbreaks. Clustering of ADSRRS fingerprinting data matched pulsed field gel electrophoresis data. The features of ADSRRS fingerprinting technique is discussed in comparison with conventional methods. Data presented here demonstrate the complexity of the epidemiological situation concerning VREM that may occur in a single medical ward. PMID- 12531504 TI - Production of amylase by newly isolated moderate halophile, Halobacillus sp. strain MA-2. AB - Production of extracellular amylase was demonstrated under stress conditions of high temperature and high salinity in aerobically cultivated culture of a newly isolated moderately halophilic bacterium of spore-forming Halobacillus sp. strain MA-2 in medium containing starch, peptone, beef extract, and NaCl. The maximum amylase production was secreted in the presence of 15% (w/v) Na(2)SO(4) (3.2 U ml(-1)). The isolate was capable of producing amylase in the presence of NaCl, NaCH(3)COOH, or KCl, with the results NaCl>NaCH(3)COOH>KCl. Maximum amylase activity was exhibited in the medium containing 5% (w/v) NaCl (2.4 U ml(-1)). Various carbon sources induced enzyme production. The potential of different carbohydrates in the amylase production was in the order: dextrin>starch>maltose>lactose>glucose>sucrose. In the presence of sodium arsenate (100 mM), maximum production of the enzyme was observed at 3.0 U ml(-1). Copper sulfate (0.1 mM) decreased the amylase production considerately, while lead nitrate had no significant enhancement on amylase production (p<0.05). The pH, temperature, and aeration optima for enzyme production were 7.8, 30 degrees C, and 200 rpm, respectively, while the optimum pH and temperature for enzyme activity was 7.5-8.5 and 50 degrees C, respectively. PMID- 12531505 TI - Development of a high throughput Pseudomonas aeruginosa epithelial cell adhesion assay. AB - Pseudomonas aeruginosa colonizes the lungs of cystic fibrosis and mechanically ventilated patients by binding to specific carbohydrate residues on the surface of lung epithelial cells. Studies have shown that blocking this interaction may have therapeutic effects in vivo. To test compounds that may have an effect on the binding of P. aeruginosa to epithelial cells, we have developed a pseudomonal adhesion assay that is compatible with high throughput technology. This assay utilizes a 96-well culture plate assay and P. aeruginosa strains that have been modified to bioluminesce. This method has proven to be a rapid, sensitive and reproducible system for screening agents that inhibit bacterial adhesion. PMID- 12531506 TI - ATP as a biomarker of viable microorganisms in clean-room facilities. AB - A new firefly luciferase bioluminescence assay method that differentiates free extracellular ATP (dead cells, etc.) from intracellular ATP (viable microbes) was used to determine the viable microbial cleanliness of various clean-room facilities. For comparison, samples were taken from both clean-rooms, where the air was filtered to remove particles >0.5 microm, and ordinary rooms with unfiltered air. The intracellular ATP was determined after enzymatically degrading the sample's free ATP. Also for comparison, cultivable microbial populations were counted on nutrient-rich trypticase soy agar (TSA) plates. Both the cultivable and ATP-based determinations indicate that the microbial burden was lower in clean-room facilities than in ordinary rooms. However, there was no direct correlation between the two sets of measurements because the two assays measured very different populations. A large fraction of the samples yielded no colony formers on TSA, but were positive for intracellular ATP. Subsequently, genomic DNA was isolated directly from selected samples and 16S rDNA fragments were cloned and sequenced, identifying nearest neighbors, many of which are known to be noncultivable in the media employed. It was concluded that viable microbial contamination can be reliably monitored by measurement of intracellular ATP, and that this method may be considered superior to cultivable colony counts due to its speed and its ability to report the presence of viable but noncultivable organisms. When the detection of nonviable microbes is of interest, the ATP assay can be supplemented with DNA analysis. PMID- 12531507 TI - A comparative study of carboxyfluorescein diacetate and carboxyfluorescein diacetate succinimidyl ester as indicators of bacterial activity. AB - Staining bacteria with esterified fluorogenic substrates followed by flow cytometric analysis offers a means for rapid detection of metabolically active bacteria. Flow cytometry (FCM) was used to assess carboxyfluorescein diacetate (CFDA) and carboxyfluorescein diacetate succinimidyl ester (CFDA/SE) as indicators of bacterial activity for cultured bacteria, including Aeromonas hydrophila, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus epidermidis and bacteria from environmental waters. In theory, CFDA/SE should be a better indicator of metabolic bacterial activity compared to CFDA due to greater intracellular retention of the fluorescent product. Qualitative and quantitative analysis of exponential phase cultures, mixtures of active and inactive cells and bacteria from environmental waters revealed CFDA was successful in detecting active bacteria, whereas CFDA/SE was not. CFDA/SE labelled inactive cells with intensities equal to that of the active population and could not even discriminate between bacteria in exponential phase growth and a fixed cell preparation. We propose that the specific mode of action of the succinimidyl ester (SE) group in combination with the nonenzymatic aqueous hydrolysis of the CFDA moiety results in the nonspecific labelling of all cells, irrespective of their metabolic state. This study shows that CFDA/SE is a poor marker of bacterial activity. PMID- 12531508 TI - Removal of PCR inhibitors from soil DNA by chemical flocculation. AB - Extracting high-purity DNA directly from soil has become essential for the study of microorganisms in environmental samples. However, many soils contain compounds that inhibit enzymes involved in manipulating DNA. In this study, chemical flocculation using multivalent cations was investigated as a potential method for eliminating soil-based inhibitors during the extraction process. The addition of AlNH(4)(SO(4))(2) during extraction significantly reduced the co-purification of PCR inhibitors with minimal loss of DNA yield. PMID- 12531509 TI - A rapid method for RNA preparation from Gram-positive bacteria. AB - We have developed a rapid method for preparation of RNA from various Gram positive bacteria. Unlike some methods, this method does not require lysozyme and proteinase K. Instead, we used glass beads to break cells more efficiently. Using this method, we successfully isolated the total RNA from various Gram-positive bacteria. This method is rapid, simple, and more economic when compared to the previously reported ones. Prepared RNA can be used for the transcriptional analysis of various Gram-positive bacteria. PMID- 12531510 TI - Insoluble TATA-binding protein accumulation in Huntington's disease cortex. AB - Huntington's disease is a dominantly inherited neurological disorder where specific neurodegeneration is caused by an extended polyglutamine stretch in the huntingtin protein. Proteins with expanded polyglutamine regions have the ability to self-aggregate and previous work in our laboratory, and by others, revealed sparse amyloid-like deposits in the Huntington's disease brain, supporting the hypothesis that the polyglutamine stretches may fold into regular beta-sheet structures. This process of folding has similarities to other neurodegenerative disorders including Alzheimer's disease, Parkinson's disease, and the prion diseases which all exhibit beta-sheet protein accumulation. We were therefore interested in testing the hypothesis that TATA-binding protein may play a role in Huntington's disease as it contains an elongated polymorphic polyglutamine stretch that ranges in size from 26 to 42 amino acids in normal individuals. A proportion of TBP alleles fall within the range of glutamine length that causes neurodegeneration when located in the huntingtin protein. In this study the distribution and cellular localisation of TATA-binding protein was compared to the distribution and cellular localisation of the huntingtin protein in the middle frontal gyrus of Huntington's disease and neurologically normal subjects. Seven different morphological forms of TATA-binding protein-positive structures were detected in Huntington's disease but not in control brain. TATA-binding protein labelling was relatively more abundant than huntingtin labelling and increased with the grade of the disease. At least a proportion of this accumulated TBP exists as insoluble protein. This suggests that TBP may play a role in the disease process. PMID- 12531511 TI - Differential expressions of mPer1 and mPer2 mRNAs under a skeleton photoperiod and a complete light-dark cycle. AB - To investigate the molecular mechanisms in photo-entrainment of circadian rhythms, expression patterns of mouse period1 (mPer1) and mPer2 mRNAs were examined in the suprachiasmatic nucleus (SCN) and skin under a complete light dark (LD) cycle and a corresponding skeleton photoperiod (SPP). Under the SPP, the robust daily expressions of mPer1 and mPer2 were maintained in the SCN and in the skin. A slower rise time for mPer1 mRNA expression was observed in the SPP compared with that in the LD condition both in the SCN and in the skin, although the expression pattern of mPer2 was not significantly different between the two lighting conditions. The results suggest that the expressions of mPer1 and mPer2 mRNAs are differentially regulated not only in the SCN but also in peripheral tissues. Furthermore, we propose that the persistent expression of mPer2 mRNA is required for the entrainment to the SPP. PMID- 12531512 TI - Characterization of mGluR5R, a novel, metabotropic glutamate receptor 5-related gene. AB - We report here the isolation of a novel gene termed mGluR5R (mGluR5-related). The N-terminus of mGluR5R is highly similar to the extracellular domain of metabotropic glutamate receptor 5 (mGluR5) whereas the C-terminus bears similarity to the testis-specific gene, RNF18. mGluR5R is expressed in the human CNS in a coordinate fashion with mGluR5. Although the sequence suggests that mGluR5R may be a secreted glutamate binding protein, we found that when expressed in HEK293 cells it was membrane associated and not secreted. Furthermore, mGluR5R was incapable of binding the metabotropic glutamate receptor class I selective agonist, quisqualate. Although mGluR5R could not form disulfide-mediated covalent homodimers, it was able to form a homomeric complex, presumably through noncovalent interactions. mGluR5R also formed noncovalent heteromeric associations with an engineered construct of the extracellular domain of mGluR5 as well as with full-length mGluR5 and mGluR1alpha. The ability of mGluR5R to associate with mGluR1alpha and mGluR5 suggests that it may be a modulator of class I metabotropic glutamate receptor function. PMID- 12531513 TI - Glutathione peroxidase inhibits cell death and glial activation following experimental stroke. AB - Stroke is a leading cause of morbidity and mortality in major industrial countries. Many factors contribute to the cellular damage resulting from ischemia reperfusion (I-R). Growing evidence indicates that reactive oxygen species (ROS) contribute significantly to this process, though their exact mechanism of action is mostly unknown. We have examined the mechanism of protection against I-R injury in transgenic mice that overexpress human glutathione peroxidase (hGPx1), using a focal cerebral I-R model. In this model, transgenic animals show significant reduction of necrotic as well as apoptotic cell death in vulnerable brain regions as demonstrated by TUNEL staining, DNA laddering and ELISA assays. We also observed decreased astrocytic and microglial activation in ischemic brains of animals overexpressing hGPx1. In wild-type mice, neuronal cell death was accompanied with compromise of vascular integrity, edema and neutrophil infiltration, whereas GPx1 mice revealed significant preservation of tissue structure and decreased infiltration of acute inflammatory cells. These results indicate that glutathione peroxidase-sensitive ROS play an important role in regulation of cell death during cerebral I-R as well as in brain inflammatory reactions. PMID- 12531514 TI - Up-regulation of mitogen-activated protein kinases ERK1/2 and MEK1/2 is associated with the progression of neurofibrillary degeneration in Alzheimer's disease. AB - The abnormal hyperphosphorylation of tau in Alzheimer's disease (AD) has been proposed to involve the extracellular-signal-regulated protein kinase (ERK) of the mitogen-activated protein (MAP) kinase family. ERK is phosphorylated and thereby activated by MAP kinase kinase (MEK). In the present study, we determined the intracellular and regional distribution of the active forms of both MEK1/2 and ERK1/2, i.e. p-MEK1/2 and p-ERK1/2 in the entorhinal, hippocampal, and temporal cortices of 49 brains staged for neurofibrillary changes according to Braak and Braak's protocol. We found that p-MEK1/2 and p-ERK1/2 were present in the initial stages of neurofibrillary degeneration in the projecting neurons in the transentorhinal region, and extended into other brain regions co-incident with the progressive sequence of neurofibrillary changes up to and including Braak stage VI. It appeared that the accumulation of p-MEK1/2 and p-ERK1/2 was initiated in the cytoplasm of pretangle neurons in varying size granules, which grew into large aggregates co-existing with the progressive development of neurofibrillary tangles. Accumulation of p-MEK1/2 and p-ERK1/2 was found in cases with stages I-III neurofibrillary degeneration, which were devoid of amyloid deposition. These data provide direct in situ evidence consistent with the possible involvement of MAP kinase pathway in the hyperphosphorylation of tau and the presence of this lesion before deposition of beta-amyloid in AD. PMID- 12531515 TI - Differential display analysis of presenilin 1-deficient mouse brains. AB - Missense mutations in presenilin 1 (PS1) gene are the most common cause of early onset familial Alzheimer's disease (FAD). AD pathogenic PS1 mutations result in elevated gamma-secretase cleavage of APP and diminished S3-site cleavage of Notch. We have previously described a PS1-hypomorphic mouse line that could survive postnatally with markedly reduced gamma-secretase cleavage of APP and S3 site cleavage of Notch, resulting in a Notch developmental phenotype similar to PS1-null mice. This model was exploited to identify genes whose expression is altered due to the loss of PS1. A global gene expression study by differential display was performed on whole brains of PS1-hypomorphic mice and their wild type siblings. In total, more than 16,000 bands corresponding to cDNAs were compared between the mutant and wild-type brains. This analysis identified 19 cDNAs showing significantly altered expression resulting from PS1 deficiency. Four of the identified cDNAs corresponded to genes that could be associated with AD or presenilin function. Hypoxia inducible factor 1a (Hif1a), NPRAP (delta-catenin) and cell division cycle 10 (CDC10) showed significantly reduced expression in the PS1-hypomorphic compared to wild-type brains, whereas expression of nucleoside diphosphate kinase sub-unit A (NDPK-A) was markedly elevated in the respective brains. Clarification of the possible role of these genes in AD and the basis for their differential expression induced by PS1-deficiency may provide insight into the disease, presenilin function and consequences of its loss, as well as possible deleterious effects of AD therapeutics aimed at inhibiting PS1. PMID- 12531516 TI - The anti-dementia drug FK960 stimulates glial glutamate release via a PKA pathway. AB - The present study was conducted to understand the mechanism underlying the facilitatory action of FK960, an anti-dementia drug, on hippocampal neurotransmission. FK960 facilitated hippocampal neurotransmission in normal mice, and also in mice lacking the glial glutamate transporter, GLT-1 (glut-1(-/ )), but to a lesser extent. FK960 enhanced glutamate release from cultured hippocampal astrocytes from normal rats and mice, while the drug had no effect on the release from cultured rat hippocampal neurons. The glutamate release was still obtained with cultured hippocampal astrocytes from glut-1(-/-) mice, suggesting that the release is not due to GLT-1-mediated counter transport of glutamate. The FK960 action was inhibited by H-89, a selective inhibitor of cAMP dependent protein kinase (PKA), bafilomycin A1, an inhibitor of vesicular transport, or BAPTA-AM, a chelator of intracellular Ca(2+). FK960 caused an increase in intracellular Ca(2+) concentrations by stored Ca(2+) release in cultured rat hippocampal astrocytes, and H-89 abolished the increase. Forskolin, a PKA activator, mimicked the effect of FK960 on intracellular Ca(2+) mobilizations. Taken together, it appears that FK960 stimulates glutamate release from astrocytes, likely as a result of raising intracellular Ca(2+) concentrations via a PKA pathway. The FK960 action would increase synaptic glutamate concentrations, in part responsible for the facilitation of hippocampal neurotransmission. The results of the present study may provide a new idea that agents targeting astrocytes could serve as anti-dementia drugs. PMID- 12531517 TI - Voltage-gated Ca2+ channel Ca(V)1.3 subunit expressed in the hair cell epithelium of the sacculus of the trout Oncorhynchus mykiss: cloning and comparison across vertebrate classes. AB - Full-length sequence (>6.5 kb) has been determined for the Ca(V)1.3 pore-forming subunit of the voltage-gated Ca(2+) channel from the saccular hair cells of the rainbow trout (Oncorhynchus mykiss). Primary structure was obtained from overlapping PCR and cloned fragments, amplified by primers based on teleost, avian, and mammalian sources. Trout saccular Ca(V)1.3 was localized to hair cells, as evidenced by its isolation from an epithelial layer in which the hair cell is the only intact cell type. The predicted amino acid sequence of the trout hair cell Ca(V)1.3 is approximately 70% identical to the sequences of avian and mammalian Ca(V)1.3 subunits and shows L-type characteristics. The trout hair cell Ca(V)1.3 expresses a 26-aa insert in the I-II cytoplasmic loop (exon 9a) and a 10 aa insert in the IVS2-IVS3 cytoplasmic loop (exon 30a), neither of which is appreciably represented in trout brain. The exon 9a insert also occurs in hair cell organs of chick and rat, and appears as an exon in human genomic Ca(V)1.3 sequence (but not in the Ca(V)1.3 coding sequence expressed in human brain or pancreas). The exon 30a insert, although expressed in hair cells of chick as well as trout, does not appear in comparable rat or human tissues. Further, the IIIS2 region shows a splice choice (exon 22a) that is associated with the hair cell organs of trout, chick, and rat, but is not found in human genomic sequence. The elucidation of the primary structure of the voltage-gated Ca(2+) channel Ca(V)1.3 subunit from hair cells of the teleost, representing the lowest of the vertebrate classes, suggests a generality of sensory mechanism for Ca(V)1.3 across hair cell systems. In particular, the exon 9a insert of this channel appears to be the molecular feature most consistently associated with hair cells from fish to mammal, consonant with the hypothesis that the latter region may be a signature for the hair cell. PMID- 12531519 TI - mRNA distribution analysis of human TRPC family in CNS and peripheral tissues. AB - The mammalian homologues of the Drosophila transient receptor potential (TRP) channel are plasma membrane proteins involved in the regulation of cellular Ca(2+) influx. These ion channels can be activated subsequent to either depletion of Ca(2+) from internal stores or through receptor-mediated processes. The mRNA expression patterns of several individual mammalian short transient receptor potential channels (TRPCs) have been described. Cross-comparisons between these data, however, are at best difficult predominantly due to the non-quantitative methods used. Furthermore there is limited data on the expression of TRPC family members in human tissues. In the present study we used a single technique, namely TaqMan real-time quantitative RT-PCR, to investigate the mRNA distribution of human TRPC1, TRPC3, TRPC4, TRPC5, TRPC6 and TRPC7 (hTRPCs) in discrete human brain areas, peripheral tissues as well as a panel of cell-lines. All hTRPCs studied were widely expressed within CNS and significant peripheral expression was often observed. Despite this, each channel exhibited a distinctive hallmark distribution profile. hTRPC1 was widely expressed in CNS and peripheral tissues, whereas hTRPC3 and hTRPC5 were predominantly expressed in tissues of CNS. hTRPC4 mRNA was detected in CNS and certain peripheral tissues such as bone, heart and prostate. hTRPC6 was homogeneously expressed throughout the CNS and peripheral tissues with the highest levels in placenta and lung. hTRPC7 mRNA was also broadly expressed in CNS as well as some peripheral tissues. The pattern of expression of the TRPCs was quite different in the various cell lines examined. TRPC3 and TRPC6 were selectively present in HEK-293 cells whilst TRPC1 was broadly distributed in the cell lines analyzed. In contrast TRPC4 and TRPC5 mRNAs were predominantly expressed in HK-2 and HEK-293 cell lines respectively. TRPC7 was selectively expressed in COS-1, COS-7 and HK-2 cell lines. These results show tissue- and cell-specific co-expression of multiple TRPC forms indicating widespread potential for formation of heteromeric channels. These data will be useful in the complex task of relating channel subunit composition to function in native cells. PMID- 12531518 TI - Estrogen receptor-beta regulates transcript levels for oxytocin and arginine vasopressin in the hypothalamic paraventricular nucleus of male mice. AB - Estrogen receptor (ER)-beta, unlike ER-alpha, is localized in the hypothalamic paraventricular nucleus (PVN) which also contains neuropeptide synthesizing neurons, such as oxytocin (OT), arginine vasopressin (AVP) and corticotropin releasing hormone (CRH). Although it is known that some ER-beta containing neurons co-express OT and AVP, but not CRH, in the PVN, it is not yet determined whether ER-beta activation may indeed play a role in estrogenic regulation on syntheses of these neuropeptides. In the present study, we tested this hypothesis by comparing the effects of estrogen on the levels of OT, AVP and CRH messenger RNA (mRNA) between ER-beta knockout (betaERKO) and wild type (WT) control male mice. Mice were gonadectomized and implanted with either a pellet containing estradiol benzoate (2.5-5.0 microg/day) or a placebo pellet for 21 days. In situ hybridization histochemistry revealed that estrogen treatment resulted in a significant increase in OT transcripts (151.6+/-6.0%) and a decrease in AVP transcripts (77.8+/-5.2%) in the PVN of WT mice, compared to the placebo control group. This estrogenic regulation of OT and AVP mRNA levels in the PVN was completely abolished in betaERKO mice. Similar genotype differences in the effects of estrogen on the numbers of OT- and AVP-containing cells were found in immunocytochemical studies performed in a separate set of mice. On the other hand, the expression of CRH mRNA in the PVN was not affected by estrogen treatment in either WT or betaERKO mice. Furthermore, estrogen did not cause any changes in the levels of OT or AVP mRNA, regardless of genotype, in the supraoptic nucleus where, unlike in rats, ER-beta containing neurons are rarely found in mice. Finally, estrogen significantly increased OT mRNA levels in both betaERKO and WT in the medial preoptic area, which contains both ER-alpha and ER beta. These results suggest that ER-beta activation may play a critical role in estrogenic regulation of OT and AVP gene expression in the PVN. PMID- 12531520 TI - Construction of recombinant pseudorabies viruses optimized for labeling and neurochemical characterization of neural circuitry. AB - In this study we have modified the neuroinvasiveness of pseudorabies virus strain Bartha, a commonly utilized trans-synaptic tract-tracer. In addition, we sought to facilitate detection of cellular mRNAs in neurons infected with the virus. In order to modify spreading characteristics, we inserted the lacZ or the GFP (green fluorescent protein) genes into the genomic loci containing the putative latency associated transcript promoter (P(LAT2)), resulting in the disruption of the promoter function. Following rat kidney injection, mutant viruses labeled central autonomic neurons in a slower and much more restricted manner than the parent Bartha strain. Since both reporter genes were controlled by the human cytomegalovirus immediate early (IE) 1 promoter, they exhibited IE expression kinetics. This property proved to be important for the co-detection of reporter proteins with neuronal mRNAs, readily detected at early but not at late stage of infection, as shown in tyrosine-hydroxylase expressing A5 catecholaminergic neurons and in serotonin transporter expressing raphe magnus neurons. PMID- 12531521 TI - Differences in the activation of the GFAP gene promoter by prion and viral infections. AB - The expression of glial fibrillary acidic protein (GFAP), a component of astroglial intermediate filaments, is regulated under developmental and pathological conditions. After surgical injury or viral infections, an increase in this protein reflects reactive gliosis in the brain. We analyzed the activation of the GFAP gene in transgenic mice using a prion and two different viruses (rabies and Theiler viruses). Inoculation of the transgenic mice with the C506M3 mouse prion strain resulted in activation of the GFAP-lacZ transgene. Expression of the GFAP transgene increased concomitantly with the expression of GFAP in astrocytes from the infected mice. In contrast, infection with rabies or Theiler's virus had no effect on the expression of the GFAP transgene, showing that the glial reactions to these infectious agents involved different mechanisms. These findings indicate that the activation of the endogenous GFAP gene as a consequence of viral infection could involve different regulatory pathways than activation as a result of prion infection. The first 2 kb upstream from the start codon of the GFAP gene seems to provide enough activation domains to produce efficient activation of the reporter gene in prion-infected mice. PMID- 12531522 TI - Studies of the pathophysiological mechanisms in frontotemporal dementia by proteome analysis of CSF proteins. AB - Comparative proteomic analysis of cerebrospinal fluid (CSF) proteins was employed for studies of the pathophysiological mechanisms in frontotemporal dementia (FTD). Two-dimensional gel electrophoresis and mass spectrometry were used for clinical screening of disease-influenced CSF proteins in 15 FTD patients compared to 12 controls. Six proteins were significantly altered in FTD compared to controls, including granin-like neuroendocrine precursor (proSAAS), pigment epithelium derived factor (PEDF), retinol-binding protein (RBP), apoE, haptoglobin, and albumin. The levels of ProSAAS, PEDF, and RBP have not been shown earlier to be involved in the FTD pathology. Recently, we have also used proteomic analysis for studies of disease-influenced CSF proteins in Alzheimer's disease (AD) patients. The most clearly affected CSF proteins were the apolipoproteins in AD, compared to controls and FTD patients. ApoE seems to be influenced to a lesser degree in FTD compared to AD. Our data showed that several proteins involved in FTD pathology are not influenced in the CSF of AD patients, and vice versa, establishing differences in the pathophysiological mechanisms between FTD and AD, two of the most common neurodegenerative disorders. PMID- 12531523 TI - Activation of protein kinase A induces neuronal differentiation of HiB5 hippocampal progenitor cells. AB - Cyclic AMP-dependent protein kinase (PKA) signaling has been shown to be a critical regulator for neuronal or glial differentiation in the developing brain and several neuronal cell lines. However, the involvement of the PKA signaling cascade in hippocampal neuronal development and differentiation is poorly understood. The present study was performed to investigate whether activation of the PKA pathway directly regulates differentiation of hippocampal progenitor cell line, HiB5. Treatment of hippocampal HiB5 cells with 0.5 mM dibutyryl-cyclic AMP (dbcAMP) at 39 degrees C in N2 medium caused dramatic morphological changes including neurite outgrowth within 24 h and an inhibition of proliferation. During these processes, PKA activity as well as phosphorylation of the cAMP responsive element binding protein (CREB) were augmented. To characterize dbcAMP induced differentiation of HiB5 cells, the expressions of several neuronal marker genes were investigated. After 24 h of dbcAMP treatment, the expression of NF-H and NF-M neuronal makers increased with a concomitant decrease in nestin (a marker for neural precursor cells) and GFAP an astrocyte marker expression, suggesting that HiB5 cells can develop a neuronal phenotype. Using the doxycycline-inducible, enhanced GFP-fused PKA catalytic subunit alpha (PKAcalpha EGFP) overexpression system, we found that overexpressed PKAcalpha-EGFP induces neurite outgrowth in HiB5 cells. Taken together, these pharmacological and genetic transfection studies provide compelling evidence for the role of PKA activation on neuronal differentiation in HiB5 hippocampal progenitor cells. PMID- 12531524 TI - Definition of pre- and postsynaptic forms of the CT carbohydrate antigen at the neuromuscular junction: ubiquitous expression of the CT antigens and the CT GalNAc transferase in mouse tissues. AB - At the rodent neuromuscular junction, the synaptic expression of the CT carbohydrate antigens is defined by the binding of two monoclonal antibodies, CT1 and CT2. CT1 preferentially stains the presynaptic membrane, while CT2 preferentially stains the postsynaptic apparatus. Here we show that the differential subsynaptic distribution of these antigens is due to a preference of CT1 for structures containing N-acetyl neuraminic acid (NeuAc) and a preference of CT2 for structures containing N-glycolyl neuraminic acid (NeuGc). This was found to be the case both in binding to cultured myotubes, where NeuAc/NeuGc levels were manipulated by feeding acetylated N-acetyl mannosamine precursors, and in binding to purified GM2 ganglioside containing either NeuAc or NeuGc. At human neuromuscular junctions, where the enzymatic machinery to make NeuGc is absent [Proc. Natl. Acac. Sci. USA 95 (1998) 11751], CT1 and GM2(NeuAc) antibodies stained, while CT2 did not. Thus, the N-glycolyl modification of sialic acid helps to define the differential distribution of the CT antigens at the rodent neuromuscular junction, and this difference is lost in humans. In addition, sulfatase and 9-O-acetylesterase treatment of cells or tissues increased the amount of CT1 and CT2 antibody binding, with sulfatase differentially unmasking CT antigen expression on particular glycoproteins. Despite its uniquely synaptic localization in skeletal muscle, the CT antigens and the CT GalNAc transferase are ubiquitously expressed in other mouse tissues, including brain, spinal cord, and peripheral nerve. One of the proteins that can be co-purified with a CT-reactive glycoprotein is alpha dystroglycan. These data better define the sub-synaptic structures of the CT carbohydrate antigens at the neuromuscular junction and demonstrate their ubiquitous presence in mouse tissues, including the brain. PMID- 12531525 TI - Effects of mutations in the highly conserved DRY motif on binding affinity, expression, and G-protein recruitment of the human angiotensin II type-2 receptor. AB - The signaling pathways for the seven transmembrane G-protein coupled angiotensin II receptors (AT(1) and AT(2)) are just beginning to be understood. While these receptors play an important role in the development and differentiation of many tissues, including the cardiovascular and central nervous systems, information about amino acid motifs involved in angiotensin II-mediated signaling is only available for the AT(1) receptor subtype. In the present study, we mutated the conserved DRY(141-143) motif in the AT(2) receptor, which is thought to be involved in G-protein recruitment. Expression of wild type and mutant receptors in CHO-K1 cell plasma membranes was confirmed using radioligand binding analyses. Our findings indicate a significant change in the binding affinities (kD) and capacities (B(max)) of the mutant receptors relative to wild type. Alanine substitutions of D(141) and DRY(141-143) resulted in a significant decrease of binding affinity for both Sar(1)Ile(8)-angiotensin II (SarIle-Ang II) (mixed agonist/antagonist) and angiotensin II (agonist). The binding affinities following alanine substitutions of R(142) and Y(143) were not significantly different from wild type receptor. Interestingly, the R(142)-A and Y(143)-A mutants revealed a significant decrease in binding levels from wild type with SarIle-Ang II, but not angiotensin II. The effect of GTPgammaS on angiotensin II binding affinity between wild type and mutant receptors was similarly significant. The D(141)-A, Y(143)-A, and DRY(141-143)-AAA mutant receptors showed a marked decrease in GTPgammaS-induced angiotensin II affinity shift. The R(142) A GTPgammaS binding affinity shift was not different from the wild type receptor. Our results support the hypothesis that the DRY motif plays a significant role in the binding affinity, structural stability and G-protein recruiting of the AT(2) receptor. PMID- 12531526 TI - Identification and characterization of a novel splice variant of the metabotropic glutamate receptor 5 gene in human hippocampus and cerebellum. AB - The G-protein coupled metabotropic glutamate receptor mGlu5 plays a pivotal role as a modulator of synaptic plasticity, ion channel activity and excitotoxicity. Two splice variants, hmGlu5a and -5b have been reported previously. During screening of a human brain cDNA library for hmGlu5a, we identified a novel variant (hmGlu5d) generated by alternative splicing at the C-terminal domain. The predicted hmGlu5d protein has a C-terminal 267 amino acid shorter than that of hmGlu5a. The pattern of mRNA expression of mGluR5 variants in human brain were analyzed by RT-PCR and in situ hybridization histochemistry. RT-PCR analysis demonstrated the presence of the hmGlu5d transcript, although at low level, in human whole brain, cerebellum, cerebral cortex and hippocampus. [3H]Quisqualate displayed similar affinity at the hmGlu5 splice variants (K(D) values of 80+/-8 and 54+/-17 nM for hmGlu5a and -5d receptors, respectively). For the five mGlu agonists studied, a similar rank order of potency was observed on both hmGlu5a and -5d receptors: quisqualate>glutamate>DHPG>L-CCGI approximately ACPD. MPEP inhibited the glutamate (2 microM)-induced [Ca(2+)](i) response in hmGlu5a and 5d-HEK293 cells also with similar potency (IC(50) values 25+/-1.5 and 20+/-1.4 nM, respectively). Therefore, the large truncation of the C-terminal tail of mGlu5 does not have any apparent major effect on the potency and efficacy of agonists as measured by the [Ca(2+)](i) responses or by activation of recombinant G-protein coupled inwardly rectifying K(+) (GIRK) channel currents. The only major functional difference is the increased sensitivity of hmGlu5d to protein kinase C (PKC)-mediated desensitization, relative to hmGlu5a. PMID- 12531527 TI - Characterization of the molecular events following impairment of NF-kappaB-driven transcription in neurons. AB - Nuclear factor-kappaB (NF-kappaB) is a transcription factor with a pivotal role in neuronal homeostasis. Indeed, NF-kappaB trans-activates several antiapoptotic genes in neurons and inhibition of NF-kappaB transcriptional activity triggers neuronal apoptosis. However, the exact mechanisms by which neurons undergo apoptosis in conditions of NF-kappaB inhibition are poorly understood. To further clarify how NF-kappaB operates in neurons, and to gather information on the molecular events occurring during NF-kappaB inhibition-dependent neuronal apoptosis, this study evaluated the effects of recently identified NF-kappaB inhibitors such as parthenolide, SN50, BAY 11-7082 and helenalin on primary cultures of rat cortical neurons. Data show that NF-kappaB was constitutively activated in neurons, and demonstrate for the first time that drug-dependent NF kappaB inhibition induced rapid mitochondrial release of cytochrome c, caspase-9 and -3 activation, poly(ADP-ribose) polymerase-1 cleavage, membrane blebbing and nuclear fragmentation, without evidence of procaspase-8 and Bid processing. Interestingly, a burst of Akt activation occurred in neurons exposed to NF-kappaB inhibitors. These events were preceded by selective reduction of mRNAs of NF kappaB-dependent, antiapoptotic Bcl-2 family members such as Bcl-x(L), Bcl-2 and, in particular, A1/Bfl-1. The present study reports a novel, detailed temporal analysis of the molecular events following impairment of NF-kappaB-driven transcription in neurons and demonstrates that inhibition of constitutive neuronal NF-kappaB activity triggers selective activation of the intrinsic apoptotic program. PMID- 12531528 TI - Neuroprotective effects of copper/zinc-dependent superoxide dismutase against a wide variety of death-inducing stimuli and proapoptotic effect of familial amyotrophic lateral sclerosis mutations. AB - Superoxide dismutase plays a key role in cell protection against the damaging effects of superoxide. Mutations in the copper/zinc dependent intracellular form of superoxide dismutase (SOD1) are associated with a subset of cases of familial amyotrophic lateral sclerosis (FALS). In this study we have investigated the effects of over-expressing wild-type SOD1 and two mutant forms of SOD1 found in FALS, G93A and G93R, on cell survival using stably transfected neuronal cells. G93R is associated with early age of onset and severely reduced erythrocyte SOD1 enzyme activity. Overexpression of wild-type SOD1 in ND7 cells significantly enhanced cell survival and reduced apoptosis after serum deprivation. Conversely, cells expressing the G93R mutation of SOD1 exhibited significantly increased cell death and increased number of TUNEL-positive cells, having a more profound effect than G93A SOD1 expressing cells, thus reflecting the relative clinical severity of these mutations. The effects of three further apoptotic and nonapoptotic death inducing paradigms were investigated, hypoxia with reperfusion, staurosporine and gamma-interferon induced cell death. With each paradigm, cell death was significantly reduced by overexpression of wild-type SOD1 and increased by overexpression of the SOD1 mutations G93A and G93R. We further used these SOD1 constructs to develop a virus expressing either wild type SOD1 or the SOD1 mutant G93R and found a similar protective effect against serum withdrawal following infection with an HSV vector expressing wild-type SOD1 which offers a potential tool for neuroprotective gene delivery in vivo. PMID- 12531529 TI - Seizure-mediated neuronal activation induces DREAM gene expression in the mouse brain. AB - Various transcriptional activators are induced in neurons concomitantly with long lasting neural activity, whereas only a few transcription factors are known to act as neural activity-inducible transcription repressors. In this study, mRNA of DREAM (DRE-antagonizing modulator), a Ca(2+)-modulated transcriptional repressor, was demonstrated to accumulate in the mouse brain after pentylenetetrazol (PTZ) induced seizures. Accumulation in the mouse hippocampus reached maximal level in the late phase (at 7-8 h) after PTZ injection. Kainic acid induced the same response. Interestingly, the late induction of DREAM expression required new protein synthesis and was blocked by MK801 suggesting that Ca(2+)-influx via NMDA receptors is necessary for the PTZ-mediated DREAM expression. In situ hybridization revealed that PTZ-induced DREAM mRNA accumulation was observed particularly in the dentate gyrus, cerebral cortex, and piriform cortex. The results of the present study demonstrate that DREAM is a neural activity stimulated late gene and suggest its involvement in adaptation to long-lasting neuronal activity. PMID- 12531530 TI - cDNA cloning and developmental expression of murine carbonic anhydrase-related proteins VIII, X, and XI. AB - To help understand the biological functions of carbonic anhydrase-related proteins (CA-RPs VIII, X, and XI), we obtained cDNA clones of murine CA-RPs X and XI and studied the tissue distribution of all three CA-RPs and their developmental expression in the murine embryonic brain. The amino acid sequences of murine CA-RPs are highly conserved with their human homologues, indicating a fundamental biological role of CA-RPs. Among a panel of vital organs, the strongest mRNA expression of all three CA-RPs was consistently observed in the brain. Immunohistochemical analysis revealed the expression of all three CA-RPs in the neural cell body and neurites. CA-RP X was also observed in the myelin sheath, as demonstrated using the shiverer demyelinated mouse. In murine embryos, CA-RP VIII and X messages first appeared in the middle of the gestation period, while the CA-RP XI message was seen at an early gestational stage and expressed to a lesser extent as gestation progressed. All CA-RPs were expressed in the neuroprogenitor cells in the subventricular area and subsequently detected in the neural cells migrating to the cortex. Although the exact function of CA-RPs is still undefined, these findings suggest an important role of CA-RPs in the CNS. PMID- 12531531 TI - Immunohistochemical investigations of neurofilament M' and alphabeta-crystallin in the magnocellular layers of the primate lateral geniculate nucleus. AB - The magnocellular and parvocellular pathways are two major processing streams in the primate visual system. Using high-density grid arrayed cDNA clones to hybridize to cDNA probes from cortical regions of each pathway, a list of candidate differentially expressed genes was produced [Mol. Brain Res. 82 (2000) 11-24]. Magnocellular pathway candidates include neurofilament M' and alphabeta crystallin. Using antibodies generated against these proteins, immunohistochemical analysis revealed preferential staining of the magnocellular layers in the primate lateral geniculate nucleus, providing verification of two candidate magnocellular-enriched genes. PMID- 12531532 TI - Differential induction and decay curves of c-fos and zif268 revealed through dual activity maps. AB - The use of inducible transcription factors for mapping neural activity is now a common procedure. We have previously developed a double-labelling technique that allows visualization of activated neurons after two different stimulation sequences. The technique exploits the differential time course of mRNA versus protein expression of transcription factors. However, the precise details of the differential time course remained unknown. Here, we provide a complete up- and downregulation profile for both the c-fos and zif268 genes, as determined through combined in situ hybridization and immunocytochemical detection of the mRNA and protein products in primary visual cortex. The data presented here can be used in the design of future studies employing double-label mapping of neural activation following a compound stimulus. PMID- 12531533 TI - Alterations in Na+/H+ exchanger and Na+/HCO3- cotransporter immunoreactivities within the gerbil hippocampus following seizure. AB - In this study, a chronological and comparative analysis of the immunoreactivities of Na(+)/H(+) exchanger 1 (NHE1), Na(+)/HCO(3)(-) cotransporter (NBC) and Na(+)/Ca(2+) exchanger (NCE) was conducted in order to identify the effects of spontaneous seizure on their protein expression levels using the gerbil model. The distribution of NHE1 and NBC immunoreactivity in the hippocampus of seizure resistant (SR) gerbils was similar to that observed in the pre-seizure group of seizure-sensitive (SS) gerbils. From 30 min to 3 h after the onset of the seizure, both NHE1 and NBC immunoreactivities were elevated in the hippocampus, as compared to the pre-seizure group of SS gerbils. At 6 h postictal, these immunoreactivities in the hippocampus had reduced to the pre-seizure level. However, NCE immunoreactivity within the hippocampus was unaltered. These findings suggest that the changes in both NHE1 and NBC immunoreactivity within the hippocampus following seizure may affect tissue excitability and play a role in the reduction of the seizure activity in the gerbil. PMID- 12531534 TI - BAD and Bcl-2 regulation are early events linking neuronal endoplasmic reticulum stress to mitochondria-mediated apoptosis. AB - The mechanism of endoplasmic reticulum (ER)-mediated apoptosis in neurons was examined. Using primary cortical neurons, we show that nordihydroguaiaretic acid (NDGA) and brefeldin A (BFA), two ER stressors, induce early ER stress as shown by Western blotting of the eukaryotic initiation factor-2alpha (eIF2alpha), an ER stress marker. This event was associated with an enhancement of neuronal apoptosis as demonstrated by the time-dependent increase in caspase-3 activity and by nuclear fragmentation. The study of the apoptotic signaling showed the translocation of cytochrome c from the mitochondrial matrix to the cytosol. Further evaluation of the apoptotic process revealed that NDGA and BFA induced a rapid dephosphorylation of BAD and decrease expression of Bcl-2. Altogether, our results indicate that neuronal ER stress is associated with an apoptotic cascade involving the mitochondria. PMID- 12531535 TI - Enhanced expression of TNF-R1 protein in NMDA-mediated cell death in the retina. AB - Multiple apoptosis-related genes are expressed in the retina after exposure to N methyl-D-aspartic acid (NMDA). For example, the mRNAs for TNF-R1, FasL, and Nur77 are up-regulated between 2.8 and 7-fold [Mol. Brain Res. 91 (2001) 34-42]. The purpose of the present study is to examine prospective changes in protein expression for these genes and to determine their cellular localizations subsequent to NMDA stimulation. Following anesthesia, a single intravitreal injection of 4 mM NMDA was administered into the right eye of anesthetized rats. The left eye was injected with phosphate-buffered saline. Retinae were harvested at 2 and 24 h postinjection. Western-blot and immunocytochemical techniques were used to detect changes in protein expression levels, and to localize their distributions within the retina. Analyses of Western blots demonstrated a significant increase in TNF-R1 (100 and 80%) compared to the sham-controls at 2 and 24 h postinjection with NMDA. Immunolabeling of TNF-R1 was observed in the inner nuclear layer (INL) at 2 h postinjection with NMDA. TNF-R1 was also clearly evident in cells within the INL and ganglion cell layers (GCL) at 24 h post injection with NMDA. In contrast to these changes in TNF-R1 there were no significant changes in the levels or distributions of FasL or Nur77 in NMDA stimulated animals at either 2 or 24 h when compared to the sham-controls. These results implicate the TNF-R1 signal transduction pathway in NMDA-induced cell death in the INL and GCL of the retina. PMID- 12531537 TI - Transbilayer phospholipid movement and the clearance of apoptotic cells. AB - When lymphocytes (and other cells) die by apoptosis, they orchestrate their own orderly removal by macrophages, and thereby prevent the inflammation that would otherwise attend cell lysis. As part of their demise, apoptotic cells disrupt the normal asymmetric distribution of phospholipids across their plasma membranes, an asymmetry normally maintained by an aminophospholipid translocase. This disruption of asymmetry, mediated by an activity known as the scramblase, generates ligands on the cell surface that trigger phagocytosis of the dying cell before lysis can occur. This crucial alteration of the plasma membrane is not dependent on caspase-mediated proteolysis, but quite unexpectedly, it is required both on the apoptotic target cell and on the phagocyte that engulfs it. At least in the phagocyte, this rearrangement may depend on the activity of an ABC ATPase, termed ABC1 in mammals and ced-7 in C. elegans. PMID- 12531538 TI - ABCA1 and the engulfment of apoptotic cells. AB - Programmed cell death is one of the major devices controlling cellular homeostasis. However, the generation of cell debris that follows the execution phase of apoptosis has to be backed up by their efficient removal by phagocyte. This highly dynamic process requires the concerted action of a number of surface molecules able to recognize early signals of membrane modifications on the apoptotic prey. Among those, the loss of phospholipid asymmetry and exposure of phosphatidylserine on the prey to be is determinant to engage phagocyte receptors and trigger the removal of corpses. A loss of membrane lipid asymmetry occurs also on the phagocyte determining its efficiency as an undertaker. Here we will discuss how, in our mind, the ATP binding cassette transporter, ABCA1, by its action on the arrangement of lipids at the phagocyte membrane, may actively promote their competence to engulf. PMID- 12531539 TI - Phospolipase A2 and apoptosis. AB - Phospolipase A(2) (PLA(2)) is the esterase activity that cleaves the sn-2 ester bond in glycerophospholipids, releasing free fatty acids and lysophospholipids. The PLA(2) activity is found in a variety of enzymes which can be divided in several types based on their Ca(2+) dependence for their activity; Ca(2+) dependent secretory phosholipases (sPLA(2)s) and cytosolic phospholipases (cPLA(2)s), and Ca(2+)-independent phospholipase A(2)s (iPLA(2)s). These enzymes also show diverse size and substrate specificity (i.e., in the fatty acid chain length and extent of saturation). Among the fatty acids released by PLA(2), arachidonic acid (AA) is of particular biological importance, because it is subsequently converted to prostanoids and leukotrienes by cyclooxygenases (COX) and lipoxygenases (LOX), respectively. Free AA may also stimulate apoptosis through activation of sphingomyelinase. Alternatively, it is suggested that oxidized metabolites generated from AA by LOX induce apoptosis. Although the precise mechanisms remain to be elucidated, changes are observed in glycerolipid metabolism during apoptotic processes. In some cells induced to undergo apoptosis, AA is released concomitant with loss of cell viability, caspase activation and DNA fragmentation. Such AA releases appear to be mediated by activation of cPLA(2) and/or iPLA(2). For example, tumor necrosis factor-alpha (TNF-alpha)-induced cell death is mediated by cPLA(2), whereas Fas-induced apoptosis appears to be mediated by iPLA(2). Some discrepancies among early experimental results were probably caused by differences in the experimental conditions such as the serum concentration, inhibitors used that are not necessarily specific to a single-type enzyme, or differential expression of each PLA(2) in cells employed in the experiments. Recent studies eliminated such problems, by carefully defining the experimental conditions, and using multiple inhibitors that show different specificities. Accordingly, more convincing data are available that demonstrate involvement of some PLA(2)s in the apoptotic processes. In addition to cPLA(2) and iPLA(2), sPLA(2)s were recently found to play roles in apoptosis. Moreover, new proteins that appear to control PLA(2)s are being discovered. Here, the roles of PLA(2)s in apoptosis are discussed by reviewing recent reports. PMID- 12531540 TI - Roles of phospholipase D in apoptosis and pro-survival. AB - Accumulating evidence has recognized phospholipase D (PLD) as an important element in signal transduction of cell responses, including proliferation and differentiation, However, its role in pro-apoptotic, anti-apoptotic or pro survival signaling is not well-understood. Involvement of PLD in these signaling mechanisms is considered to differ depending on the cell type and the extracellular stimulus. PMID- 12531541 TI - Phosphatidylcholine and cell death. AB - Phosphatidylcholine (PC) constitutes a major portion of cellular phospholipids and displays unique molecular species in different cell types and tissues. Inhibition of the CDP-choline pathway in most mammalian cells or overexpression of the hepatic phosphatidylethanolamine methylation pathway in hepatocytes leads to perturbation of PC homeostasis, growth arrest or even cell death. Although many agents that perturb PC homeostasis and induce cell death have been identified, the signaling pathways that mediate this cell death have not been well defined. This review summarizes recent progress in understanding the relationship between PC homeostasis and cell death. PMID- 12531542 TI - Cardiolipin and apoptosis. AB - Cardiolipin (CL) is recognized to be an essential phospholipid in eukaryotic energy metabolism so that physiological and pathological perturbations in its synthetic and catabolic pathways play key roles in maintaining mitochondrial structure and function, and ultimately cell survival. This review describes potential regulatory mechanisms in CL synthesis and the effects of de-acylation pathways on steady state levels of CL and its interaction with cytochrome c. The latter interaction is significant in the initiation of programmed cell death. Physiological factors that modify CL acylation include ageing, dietary influences and ischemia/reperfusion where the terminal events may be either necrosis or apoptosis. In various pathologies, phospholipase activity increases in response to production of peroxidized CL. The cell may use lysosomal or mitochondrial pathways for CL degradation. However, the manner by which CL and cytochrome c leave the mitochondria is not well understood. The lipid (CL)-bound form of cytochrome c is thought to initiate apoptosis via a lipid transfer step involving mitochondrially targeted Bid. A direct relationship between CL loss and cytochrome c release from the mitochondria has been identified as an initial step in the pathway to apoptosis. An absolute requirement for CL in the function of crucial mitochondrial proteins, e.g., cytochrome oxidase and the adenine nucleotide translocase, are likely additional factors impacting apoptosis and cellular energy homeostasis. This is reflected in the occurrence of both oncotic and apoptotic events in ischemia and reperfusion injury. Other potential clinical manifestations of perturbations of CL synthesis are discussed with particular emphasis on Barth Syndrome where a primary defect can be attributed to CL metabolism and is associated with dilated cardiomyopathy. Finally, the model of fatty acid induced apoptosis is used as a paradigm to our understanding of the temporal relationship between decreased mitochondrial CL, release of cytochrome c, and initiation of apoptosis. PMID- 12531543 TI - Lysophosphatidic acid as a novel cell survival/apoptotic factor. AB - Lysophosphatidic acid (LPA) activates its cognate G protein-coupled receptors (GPCRs) LPA(1-3) to exert diverse cellular effects, including cell survival and apoptosis. The potent survival effect of LPA on Schwann cells (SCs) is mediated through the pertussis toxin (PTX)-sensitive G(i/o)/phosphoinositide 3-kinase (PI3K)/Akt signaling pathways and possibly enhanced by the activation of PTX insensitive Rho-dependent pathways. LPA promotes survival of many other cell types mainly through PTX-sensitive G(i/o) proteins. Paradoxically, LPA also induces apoptosis in certain cells, such as myeloid progenitor cells, hippocampal neurons, and PC12 cells, in which the activation of the Rho-dependent pathways and caspase cascades has been implicated. The effects of LPA on both cell survival and apoptosis underscore important roles for this lipid in normal development and pathological processes. PMID- 12531544 TI - Ceramide in apoptosis: an overview and current perspectives. AB - Recent years have witnessed significant advances in the understanding of the role of ceramide in apoptosis. This review summarizes these recent findings and discusses insights from studies of ceramide metabolism, topology, and effector actions. The recent identification of several genes for enzymes of ceramide metabolism, the development of mass spectrometric methods for ceramide analysis, and the increasing molecular and pharmacological tools to probe ceramide metabolism and function promise an accelerated phase in defining the molecular and biochemical details of the role of ceramide in apoptosis. PMID- 12531545 TI - Sphingomyelin hydrolysis during apoptosis. AB - Sphingolipid breakdown products are now being recognized as important players in apoptosis. Ceramide, which is considered to serve as second messenger, is mainly generated by hydrolysis of the membrane sphingophospholipid sphingomyelin (SM) through the action of a sphingomyelinase (SMase). However, little is known about the localization and regulation of this phenomenon. Here, we summarize the current knowledge on the function of SM hydrolysis in apoptosis signaling. In particular, the present review focuses on the role of neutral sphingomyelinase (N SMase) in the generation of the proapoptotic ceramide. This enzyme is regulated by several mechanisms, including the tumor necrosis factor (TNF) receptor associated protein FAN (for factor associated with N-SMase activation) and oxidative stress. These observations place SMase activation and SM hydrolysis as early events in the apoptosis signaling cascade. PMID- 12531547 TI - Ceramide and cell death receptor clustering. AB - Acid sphingomyelinase (ASM) has been shown to be activated by a variety of receptor molecules and stimuli including CD95, the tumor necrosis factor receptor (TNF-R), CD40, CD28, LFA-1, CD5, during development, irradiation, heat shock, UV light or bacterial and viral infections. The central role of ASM-released ceramide in the response to those stimuli is confirmed by several genetic studies. ASM and ceramide might mediate their biological effects by the activation of several intracellular signaling molecules including cathepsin D, phospholipase A(2) or the kinase suppressor of Ras. In addition, recent fluorescence microscopy studies indicate that distinct, small membrane domains, termed rafts, are modified by ceramide to form larger domains, which serve to cluster receptor molecules. The generation of a high receptor density might be required for initiation of receptor-specific signaling and explain the function of the ASM and ceramide in multiple signaling pathways. PMID- 12531546 TI - Sphingolipids, apoptosis, cancer treatments and the ovary: investigating a crime against female fertility. AB - Premature ovarian failure and infertility are well-known side-effects observed in young girls and reproductive-age women treated for cancer. Although the need for tumor eradication in these patients is clear, the long-term consequences of chemotherapy and radiation on non-target tissues, such as the ovaries where large numbers of germ cells (oocytes) are also killed off, are substantial. Unfortunately, the mechanism mediating the undesirable toxicity of cancer therapies in the female gonads has only recently been explored. Nevertheless, some important insights into the role of ceramide and sphingosine-1-phosphate (S1P) as a mediator and suppressor, respectively, of cancer therapy-induced oocyte apoptosis have emerged over the past few years. Such findings are exciting in that a better understanding of the crime--how radiation and chemotherapy kill off this irreplaceable population of innocent cells in the ovaries--may finally allow for the development of novel lipid-based strategies to combat infertility and premature menopause in female cancer patients. PMID- 12531548 TI - Serine palmitoyltransferase: role in apoptotic de novo ceramide synthesis and other stress responses. AB - Serine palmitoyltransferase is the first and rate-limiting enzyme of sphingolipid synthesis. As such, it is a central control point in the synthesis of bioactivate sphingolipids, and it plays an important role in mediating cellular stress responses. In this review, its role in mediating these responses is discussed within the context of de novo ceramide synthesis. Furthermore, a discussion is provided of its regulation as discerned from both yeast and mammalian studies. PMID- 12531549 TI - Sphingosine in apoptosis signaling. AB - The sphingolipid metabolites ceramide, sphingosine, and sphingosine 1-phosphate contribute to controlling cell proliferation and apoptosis. Ceramide and its catabolite sphingosine act as negative regulators of cell proliferation and promote apoptosis. Conversely, sphingosine 1-phosphate, formed by phosphorylation of sphingosine by a sphingosine kinase, has been involved in stimulating cell growth and inhibiting apoptosis. As the phosphorylation of sphingosine diminishes apoptosis, while dephosphorylation of sphingosine 1-phosphate potentiates it, the role of sphingosine as a messenger of apoptosis is of importance. Herein, the effects of sphingosine on diverse signaling pathways implicated in the apoptotic process are reviewed. PMID- 12531551 TI - Glucosylceramide synthase and apoptosis. AB - Glucosylceramide synthase (GCS) is an enzyme inherent to ceramide metabolism. The enzyme catalyzes the transfer of glucose to ceramide, the first committed step in glycolipid biosynthesis. Known for many years as a branch point enzyme directing synthesis of cerebrosides and gangliosides, GCS has recently been implicated in the cytotoxic response of cancer cells to chemotherapy. With ceramide now occupying a central role in the signaling mechanisms of apoptosis, the position of GCS as sentry is perhaps not unexpected. In particular, it has been recognized that the toxic response of cells to chemotherapy is impaired when GCS activity is elevated and heightened when GCS activity is blocked. Herein we review the control points of ceramide metabolism with special regard to GCS and the cytotoxic response. PMID- 12531550 TI - Yeast sphingolipids: metabolism and biology. AB - Sphingolipids have recently emerged as important bioactive molecules in addition to being critical structural components of cellular membranes. These molecules have been implicated in regulating cell growth, differentiation, angiogenesis, apoptosis, and senescene. To study sphingolipid mediated biology, it is necessary to investigate sphingolipid metabolism and its regulation. The yeast Saccharomyces cerevisiae has allowed such studies to take place as the sphingolipid metabolic and regulatory pathways appear conserved across species. Using yeast genetic approaches most enzymes of sphingolipid metabolism have been identified and cloned which has led to identification of their mammalian homologues. Many of the yeast enzymes are targets of fungal toxins thus underscoring the importance of this pathway in yeast cell regulation. This review focuses on the yeast sphingolipid metabolic pathway and its role in regulation of yeast biology. Implication of the insights gained from yeast to mammalian cell regulation are discussed. PMID- 12531552 TI - GD3 ganglioside and apoptosis. AB - Lipid and glycolipid mediators are important messengers of the adaptive responses to stress, including apoptosis. In mammalian cells, the intracellular accumulation of ganglioside GD3, an acidic glycosphingolipid, contributes to mitochondrial damage, a crucial event during the apoptopic program. GD3 is a minor ganglioside in most normal tissues. Its expression increases during development and in pathological conditions such as cancer and neurodegenerative disorders. Intriguingly, GD3 can mediate additional biological events such as cell proliferation and differentiation. These diverse and opposing effects indicate that tightly regulated mechanisms, including 9-O-acetylation, control GD3 function, by affecting intracellular levels, localization and structure of GD3, and eventually dictate biological outcomes and cell fate decisions. PMID- 12531553 TI - Fumonisins and fumonisin analogs as inhibitors of ceramide synthase and inducers of apoptosis. AB - Sphingoid bases are growth inhibitory and pro-apoptotic for many types of cells when added to cells exogenously, and can be elevated to toxic amounts endogenously when cells are exposed to inhibitors of ceramide synthase. An important category of naturally occurring inhibitors are the fumonisins, which inhibit ceramide synthase through structural similarities with both the sphingoid base and fatty acyl-CoA co-substrates. Fumonisins cause a wide spectrum of disease (liver and renal toxicity and carcinogenesis, neurotoxicity, induction of pulmonary edema, and others), and most-possibly all-of the pathophysiologic effects of fumonisins are attributable to disruption of the sphingolipid metabolism. The products of alkaline hydrolysis of fumonisins (which occurs during the preparation of masa flour for tortillas) are aminopentols that also inhibit ceramide synthase, but more weakly. Nonetheless, the aminopentols (and other 1-deoxy analogs of sphinganine) are acylated to derivatives that inhibit ceramide synthase, perhaps as product analogs, elevate sphinganine, and kill the cells. Somewhat paradoxically, fumonisins sometimes stimulate growth and inhibit apoptosis, possibly due to elevation of sphinganine 1-phosphate, which is known to have these cellular effects. These findings underscore the complexity of sphingolipid metabolism and the difficulty of identifying the pertinent mediators unless a full profile of the potentially bioactive species is evaluated. PMID- 12531555 TI - Lipoapoptosis: its mechanism and its diseases. AB - The balance between cell division and cell death determines the cell population of an organ. When cell death exceeds cell replacement in an organ, a functional deficit is created. A metabolic cause of programmed cell death, lipoapoptosis, has recently been identified to occur in obesity and aging. If nonadipose tissues are exposed to an excess of long-chain fatty acids, unless leptin action increases their oxidation sufficiently, unoxidized fatty acids enter nonoxidative pathways. While initially they are sequestered as harmless neutral fat, ultimately some will enter more toxic pathways. One of these, the de novo ceramide pathway, has been implicated in the lipoapoptosis of beta-cells and myocardiocytes of congenitally obese rats in which leptin action is defective. Here we review the mechanisms of lipoapoptosis and the diseases that result from this cause of a diminishing cell population of these organs. We suggest that some of the components of the metabolic syndrome of obese humans and the sarcopenia of aging may be result of failure of leptin liporegulation to prevent lipid overload of lean body mass and lipoapoptosis in certain organ systems. PMID- 12531554 TI - Sphingosine kinase, sphingosine-1-phosphate, and apoptosis. AB - The sphingolipid metabolites ceramide (Cer), sphingosine (Sph), and sphingosine-1 phosphate (S1P) play an important role in the regulation of cell proliferation, survival, and cell death. Cer and Sph usually inhibit proliferation and promote apoptosis, while the further metabolite S1P stimulates growth and suppresses apoptosis. Because these metabolites are interconvertible, it has been proposed that it is not the absolute amounts of these metabolites but rather their relative levels that determines cell fate. The relevance of this "sphingolipid rheostat" and its role in regulating cell fate has been borne out by work in many labs using many different cell types and experimental manipulations. A central finding of these studies is that Sph kinase (SphK), the enzyme that phosphorylates Sph to form S1P, is a critical regulator of the sphingolipid rheostat, as it not only produces the pro-growth, anti-apoptotic messenger S1P, but also decreases levels of pro-apoptotic Cer and Sph. Given the role of the sphingolipid rheostat in regulating growth and apoptosis, it is not surprising that sphingolipid metabolism is often found to be disregulated in cancer, a disease characterized by enhanced cell growth, diminished cell death, or both. Anticancer therapeutics targeting SphK are potentially clinically relevant. Indeed, inhibition of SphK has been shown to suppress gastric tumor growth [Cancer Res. 51 (1991) 1613] and conversely, overexpression of SphK increases tumorigenicity [Curr. Biol. 10 (2000) 1527]. Moreover, S1P has also been shown to regulate angiogenesis, or new blood vessel formation [Cell 99 (1999) 301], which is critical for tumor progression. Furthermore, there is intriguing new evidence that S1P can act in an autocrine and/or paracrine fashion [Science 291 (2001) 1800] to regulate blood vessel formation [J. Clin. Invest. 106 (2000) 951]. Thus, SphK may not only protect tumors from apoptosis, it may also increase their vascularization, further enhancing growth. The cytoprotective effects of SphK/S1P may also be important for clinical benefit, as S1P has been shown to protect oocytes from radiation-induced cell death in vivo [Nat. Med. 6 (2000) 1109]. Here we review the growing literature on the regulation of SphK and the role of SphK and its product, S1P, in apoptosis. PMID- 12531556 TI - Oxidized low-density lipoprotein-induced apoptosis. AB - Cultured cells are able to oxidize low-density lipoproteins (LDL) and oxidized LDL (oxLDL), which are present in atherosclerosis areas, exhibit a variety of biological properties potentially involved in atherogenesis. This review is focused on the toxicity of oxLDL, more precisely on the toxic compounds generated during LDL oxidation, the features and the mechanisms of cell death (apoptosis or necrosis) induced by oxLDL. After internalization, toxic oxidized lipids, namely lipid peroxides, oxysterols and aldehydes, induce modifications of cell proteins, elicit oxidative stress, lipid peroxidation and alter various signaling pathways and gene expression. These events may participate in the toxic effect, and converge to trigger an intense, delayed and sustained calcium peak which elicits either apoptosis or necrosis processes. OxLDL-induced apoptosis involves both mitochondrial and death-receptor (Fas/FasL) apoptotic pathways, thereby activating the classical caspase cascade and subsequent biochemical and morphological apoptotic features. When apoptosis is blocked by overexpression of Bcl-2, oxLDL trigger necrosis through a calcium-dependent pathway. Apoptosis occurring in atherosclerotic areas is potentially involved in endothelial cell lining defects, necrotic core formation and plaque rupture or erosion which may trigger atherothrombotic events. However, the precise role of oxLDL in apoptosis/necrosis occurring in vivo in atherosclerotic plaques remains to be clarified. PMID- 12531557 TI - Mechanical induction in limb morphogenesis: the role of growth-generated strains and pressures. AB - Morphogenesis is regulated by intrinsic factors within cells and by inductive signals transmitted through direct contact, diffusible molecules, and gap junctions. In addition, connected tissues growing at different rates necessarily generate complicated distributions of physical deformations (strains) and pressures. In this Perspective we present the hypothesis that growth-generated strains and pressures in developing tissues regulate morphogenesis throughout development. We propose that these local mechanical cues influence morphogenesis by: (1) modulating growth rates; (2) modulating tissue differentiation; (3) influencing the direction of growth; and (4) deforming tissues. It is in this context that we review concepts and experiments of cell signaling and gene expression in various mechanical environments. Tissue and organ culture experiments are interpreted in light of the developmental events associated with the growth of the limb buds and provide initial support for the presence and morphological importance of growth-generated strains and pressures. The concepts presented are used to suggest future lines of research that may give rise to a more integrated mechanobiological view of early embryonic musculoskeletal morphogenesis. PMID- 12531558 TI - Directing the expression of a green fluorescent protein transgene in differentiated osteoblasts: comparison between rat type I collagen and rat osteocalcin promoters. AB - The osteocalcin (OC) and a 2.3 kb fragment of the collagen promoter (Col2.3) have been used to restrict transgenic expression of a variety of proteins to bone. Transgenic mice carrying a green fluorescent protein (GFP) gene driven by each promoter were generated. Strong GFP expression was detected in OC-GFP mice in a few osteoblastic cells lining the endosteal bone surface and in scattered osteocytes within the bone matrix in long bones from 1-day-old to 6-month-old transgenic animals. Similar findings were noted in the forming tooth in which only individual odontoblasts expressed GFP without detectable expression from the dental pulp. This limited pattern of OC-GFP-positive cells contrasts with the uniform expression in the Col2.3GFP mice in which large proportion of osteoblasts, odontoblasts, and osteocytes strongly expressed the transgene. To assess transgene expression during in vitro differentiation, marrow stromal cell and neonatal calvarial osteoblast cultures were analyzed. The activity of both transgenes was restricted to mineralized nodules but the number of positive cells was lower in the OC-GFP-derived cultures. The different temporal and spatial pattern of each transgene in vivo and in vitro reveals potential advantages and disadvantages of these two transgene models. PMID- 12531559 TI - Differentiation of murine preosteoblastic KS483 cells depends on autocrine bone morphogenetic protein signaling during all phases of osteoblast formation. AB - In this study, we examine the role of bone morphogenetic protein (BMP) signaling during differentiation of the murine preosteoblastic KS483 cell line, which formed alkaline phosphatase (ALP)-positive and mineralized nodules during a 3 week culture period. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) demonstrated the presence of various BMPs (BMP-2, -3, -4, -6, 7, and -8A and -8B), BMP type I and II receptors (ALK2, ALK3, ALK4, BMPR-II, and ActR-IIA and -IIB), BMP antagonists (DAN, gremlin, chordin, cerberus, noggin, and tsg), and Smads 1-8. mRNA expression of these genes did not change during differentiation, except for BMP-3, BMP-8a, and noggin. BMP-3 increased gradually, particularly in the matrix formation phase; BMP-8a was induced from the onset of matrix maturation and mineralization, in parallel to the expression of osteocalcin; and noggin tended to decline during the mineralization phase. Treatment of KS483 cells with the BMP antagonists noggin or soluble truncated BMPR-IA, either continuously or during distinct periods of osteoblast differentiation; that is, matrix formation or matrix maturation and mineralization phase, decreased ALP-positive and mineralized nodule area independent of the phase of osteoblast differentiation. Notably, the antagonists inhibited mineralization of already existing nodules. Similarly, BMP-4 stimulated differentiation not only at the beginning of the culture period, but also at late stages of differentiation. These data indicate that autocrine BMP signaling is involved in KS483 osteoblastic differentiation not only during the early phase of differentiation, but also during matrix maturation and mineralization. The different expression patterns of components of BMP signaling in the KS483 cells suggest distinct functions of individual BMPs during osteoblast differentiation. In summary, our data suggest that BMP activity is required not only for initiation of osteoblast differentiation and further development of early osteoblasts, but is also involved in late-stage osteoblast differentiation and matrix mineralization. PMID- 12531560 TI - Mouse genetic model for bone strength and size phenotypes: NZB/B1NJ and RF/J inbred strains. AB - The relationships of bone size, bone strength, and bone formation were investigated in two strains of mice, NZB/B1NJ and RF/J. Measurement of the femur midshaft size by peripheral quantitative computed tomography (pQCT) showed that the RF/J mice had a 32% greater cross-sectional area than NZB/B1NJ mice at 10 weeks of age, and a 38% greater cross-sectional area at 22 weeks of age. Body weight in the RF/J mice was 10% higher at 10 weeks but 9% lower at 22 weeks. Bone strength was determined by a three-point bending method. In agreement with the difference in bone cross-sectional area, the femurs of the RF/J mice were stronger (80% greater) and stiffer (80% greater) than the bones of the NZB/B1NJ mice. To determine whether periosteal bone formation played a role in the greater size of the RF/J mice, the mice were injected with tetracycline to label areas of new bone formation. Histomorphometrical analysis of the femur diaphysis demonstrated higher rates of periosteal bone formation (131% greater) and of periosteal forming surface (81% greater) in RF/J than in NZB/B1NJ mice. We conclude that a high rate of periosteal bone formation increases bone size and strength in RF/J mice when compared with NZB/B1NJ mice. The NZB/B1NJ and RF/J mice should be an excellent model to investigate the genes that regulate femur size and strength. PMID- 12531561 TI - Effects of transforming growth factor-beta deficiency on bone development: a Fourier transform-infrared imaging analysis. AB - Transforming growth factor-beta 1 (TGF-beta1) is a cytokine member of the TGF beta superfamily involved in the control of proliferation and differentiation of various cell types. TGF-beta1 plays an important role in bone formation and resorption. To determine the effect of TGF-beta1 deficiency on bone mineral and matrix, tibias from mice in which TGF-beta1 expression had been ablated (TGF beta1 null) were analyzed and compared with background- and age-matched wild-type (WT) control animals by Fourier transform-infrared imaging (FTIRI) and histochemistry. FTIRI allows the characterization of nondemineralized thin tissue sections at the ultrastructural level with a spatial resolution of approximately 7 microm. The spectroscopic parameters calculated were: mineral-to-matrix ratio (previously shown to correspond to ash weight); mineral crystallinity (related to the crystallographically determined crystallite size and perfection in the apatite c-axis direction); and collagen maturity (related to the ratio of pyridinoline:deH-DHLNL collagen cross-links). Several fields were selected to represent different stages of bone development within the same specimen from the secondary ossification center to the distal diaphysis. Anatomically equivalent areas were compared as a function of age and genotype. The spectroscopic results were expressed both as color-coded images and as pixel population distributions for each of the three parameters monitored. Based on comparisons of histochemistry and FTIRI, there were distinctive age and genotype variations. At all ages examined, in the TGF-beta1 null mice growth plates, alkaline phosphatase (ALP) activity and collagen maturity were reduced, but no effect on mineral content or crystallinity was noted. In the TGF-beta1 null mice metaphyses, there was a persistence of trabeculae, but no significant alterations in mineral content or crystallinity. In contrast, mineral content, mineral crystallinity, and collagen maturity were reduced in the secondary ossification center and cortical bone of the TGF-beta1 null mice. These results, consistent with a mechanism of impaired bone maturation in the TGF-beta1 null mice, may be directly related to TGF-beta1 deficiency and indirectly to increased expression of inflammatory cytokines in the TGFbeta1 null mice. PMID- 12531562 TI - Subclinical hypervitaminosis A causes fragile bones in rats. AB - Excessive intake of vitamin A has been associated with an increased risk of hip fracture in humans. This finding has raised the question of whether long-term intake of relatively moderate doses ("subclinical" hypervitaminosis A) contributes to fracture risk. Although it has been known for more than half a century that toxic doses of vitamin A lead to spontaneous fractures in rats, the lowest intake that induces adverse effects is not known, and the result of exposure to excessive doses that do not cause general toxicity has been rarely investigated. In this study, mature female rats were fed a standard diet with 12 IU vitamin A/g pellet (control, C), or standard diet supplemented with either 120 IU ("10 x C") or 600 IU ("50 x C") vitamin A/g pellet for 12 weeks. Fifteen animals were included in each group. The supplemented diets correspond to a vitamin A intake of approximately 1800 IU/day and 9000 IU/day, respectively. The latter dose is about one third of that previously reported to cause skeletal lesions. At the end of the study, serum retinyl esters were elevated 4- (p < 0.01) and 20-fold (p < 0.001) and the total amount of liver retinoid had increased 3- (p < 0.001) and 7-fold (p < 0.001) in the 10 x C and 50 x C group, respectively. The animals showed no clinical signs of general toxicity, and there were no significant bone changes in the 10 x C group. However, in the 50 x C group, a characteristic thinning of the cortex (cortical area -6.5% [p < 0.001]) and reduction of the diameter of the long bones were evident (bone cross sectional area -7.2% [p < 0.01] at the midshaft and -11.0% [p < 0.01] at the metaphysis), as measured by peripheral quantitative computed tomography. In agreement with these data and a decreased polar strength strain index (-14.0%, p < 0.01), the three-point bending breaking force of the femur was reduced by 10.3% (p < 0.01) in the 50 x C group. These data indicate that the negative skeletal effects appear at a subchronic vitamin A intake of somewhere between 10 and 50 times the standard diet. This level is considerably lower than previously reported. Our results suggest that long-term ingestion of modest excesses of vitamin A may contribute to fracture risk. PMID- 12531563 TI - Site-specific immunostaining for type X collagen in noncalcified articular cartilage of canine stifle knee joint. AB - Type X collagen is a short-chain collagen that is strongly expressed in hypertrophic chondrocytes. In this study, we used an immunohistochemical technique exploiting a prolonged hyaluronidase unmasking of type X collagen epitopes to show that type X collagen is not restricted to calcified cartilage, but is also present in normal canine noncalcified articular cartilage. A 30 degrees valgus angulation procedure of the right tibia was performed in 15 dogs at the age of 3 months, whereas their nonoperated sister dogs served as controls. Samples were collected 7 and 18 months after the surgery and immunostained for type X collagen. The deposition of type X collagen increased during maturation from age 43 weeks to 91 weeks. In the patella, most of the noncalcified cartilage stained for type X collagen, whereas, in the patellar surface of the femur, it was present mainly in the femoral groove close to cartilage surface. In femoral condyles, the staining localized mostly in the superficial cartilage on the lateral and medial sides, but not in the central weight-bearing area. In tibial condyles, type X collagen was often observed close to the cartilage surface in medial parts of the condyles, although staining could also be seen in the deep zone of the cartilage. Staining for type X collagen appeared strongest at sites where the birefringence of polarized light was lowest, suggesting a colocalization of type X collagen with the collagen fibril arcades in the intermediate zone. No significant difference in type X collagen immunostaining was observed in lesion-free articular cartilage between controls and dogs that underwent a 30 degrees valgus osteotomy. In osteoarthritic lesions, however, there was strong immunostaining for both type X collagen and collagenase-induced collagen cleavage products. The presence of type X collagen in the transitional zone of cartilage in the patella, femoropatellar groove, and in tibial cartilage uncovered by menisci suggests that it may involve a modification of collagen fibril arrangement at the site of collagen fibril arcades, perhaps providing additional support to the collagen network. PMID- 12531564 TI - Cancellous bone microdamage in the proximal femur: influence of age and osteoarthritis on damage morphology and regional distribution. AB - This study describes the in vivo distribution of cancellous bone microdamage in the proximal femur of an autopsy control sample. In addition, in vivo microdamage in the region medial to the greater trochanter of the proximal femur is compared between patients with severe osteoarthritis and controls. Taken at autopsy, the control group comprised normal right proximal femora that were then cut in the coronal plane with an Exakt saw (n = 12; aged 20-83 years). Cancellous bone samples were taken from the subchondral principal compressive region, the medial principal compressive region, and medial to the greater trochanter. A cancellous bone core biopsy was taken of the region medial to the greater trochanter (of the proximal femur) from patients with primary osteoarthritis undergoing total hip replacement surgery (n = 33; aged 37-85 years). Samples were embedded in resin, and in vivo microdamage identified in 70-microm-thick sections using the basic fuchsin en bloc staining technique. Microdamage was similar in all proximal femur sites in controls, except in the subchondral principal compressive region, where a significantly smaller crack length (microm) was identified (p < 0.05). In the region medial to the greater trochanter, osteoarthritic vs. control group comparisons showed that the crack density (#/mm(2)) and crack surface density (mm/mm(2)) were not significantly different, but crack length was significantly less (p < 0.03) and damage volume fraction was significantly increased for osteoarthritics (p < 0.005). The osteoarthritic and control data for crack density, and the osteoarthritic data for damage volume fraction, showed a nonlinear increase with age. Furthermore, crack length was not dependent on damage volume fraction or age for either the osteoarthritic or control group. This study identified differences in microdamage between osteoarthritic and autopsy control cases. We hypothesize that these results are consistent with the reported bone material property differences for osteoarthritis. In addition, the relatively uniform distribution of microdamage in the control group suggests that the principal components of the femoral cancellous bone network are equally exposed to deformations resulting in microdamage. Further study into the factors that influence the accumulation and skeletal distribution of microdamage is fundamental to understanding skeletal health. PMID- 12531565 TI - Patterns of osteocalcin and bone specific alkaline phosphatase by age, gender, and race or ethnicity. AB - A variety of biochemical markers of bone turnover that assess bone formation or resorption are now available for research and clinical application. However, our understanding of the usual pattern of these measures over age in the general population is limited. Therefore, values of two bone formation markers, serum osteocalcin (Oc) and bone specific alkaline phosphatase (bone ALP), were compared by age, gender, and race or ethnicity using serum obtained from a subsample of blacks, whites, and Mexican Americans from the third National Health and Nutrition Examination Survey (NHANES). In all racial and ethnic groups, mean values of both serum Oc and bone ALP were lower in women than in men <50 years old. In individuals > or =50 years of age, Oc was significantly higher in women than in men. When analyzed in these two broad age groups, Oc was lower in older black men than in white or Mexican American men, but bone ALP was not different among the groups. In women, Oc levels tended to be lower in the black women than in white or Mexican American women. In contrast, bone ALP tended to be lower in white women than in black or Mexican American women. On the other hand, when analyzed by decade, patterns differed between the two markers in both men and women. In women, both Oc and bone ALP rose postmenopausally. However, bone ALP plateaued in the sixth through eighth decades, whereas Oc levels tended to increase further. In men, Oc was highest in the 20-29 year age group, declined and stabilized, then increased again in the seventh decade. In contrast, mean bone ALP did not differ by decade in men. Our data document differences in levels of circulating Oc and bone ALP by age, gender, and race/ethnicity. The age patterns reflected by the two markers are not concordant and distinctions are most evident in the latter decades. Our findings suggest that the specific osteoblast activity reflected by these markers responds differently to the physiologic changes that occur later in life. PMID- 12531566 TI - Relationships between osteocyte density and bone formation rate in human cancellous bone. AB - Iliac cancellous osteocyte density decreases with age in deep bone but not in superficial bone, most likely because of remodeling. It has been suggested that osteocytes can inhibit bone remodeling. Accordingly, we examined the relationship between osteocyte density and bone formation rate in 92 healthy women. In superficial bone (<25 microm from the surface), we found a weak but significant (p < 0.03) inverse correlation between BFR/BS and Ot. N/B.Ar that was unaffected by menopause and independent of age. A weaker positive relationship with empty lacunar density improved significance. The data appear to suggest a negative feedback loop, but osteocytes explain only 10% of the variance in BFR/BS, and 97% of the variance in osteocyte density is explained by total lacunar density. This measure of initial osteocyte density during bone formation has a high coefficient of variation (20%) indicating large individual differences. We conclude that: (1) our data support the proposal that osteocytes can inhibit bone remodeling; (2) osteocyte density in superficial bone depends mainly on initial osteocyte density during bone formation and is maintained but not regulated by bone remodeling; and (3) the inverse relationship between BFR/BS and osteocyte density may reflect the homeostatic need to maintain calcium exchangeability in the lining cell-osteocyte syncytium. PMID- 12531567 TI - Falls explain between-center differences in the incidence of limb fracture across Europe. AB - There is important geographic variation in the occurrence of the major osteoporotic fractures across Europe. The aim of this study was to determine whether between-center variation in limb fracture rates across Europe could be explained by variation in the incidence of falls. Men and women, aged 50-79 years, were recruited from population-based registers in 30 European centers. Subjects were followed by postal questionnaire to ascertain the occurrence of incident fractures, and were also asked about the occurrence and number of recent falls. Self-reported fractures were confirmed, where possible, by review of the radiographs, medical record, or subject interview. The age- and gender-adjusted incidence of falls was calculated by center using Poisson regression. Poisson regression was also used to assess the extent to which between-center differences in the incidence of limb fractures could be explained by differences in the age- and gender-adjusted incidence of falls at those centers. In all, 6302 men (mean age 63.9 years) and 6761 women (mean age 63.1 years) completed at least one questionnaire concerning fractures and falls. During a median follow-up time of 3 years, 3647 falls were reported by men and 4783 by women. After adjusting for age and gender, there was evidence of significant between-center differences in the occurrence of falls. There was also between-center variation in the occurrence of upper limb, lower limb, and distal forearm fractures. Variation in the age- and gender-adjusted center-specific fall rates explained 24%, 14%, and 6% of the between-center variation in incidence of distal forearm and upper and lower limb fractures, respectively. Given the constraints inherent in such an analysis, in men and women aged 50-79 years, variation in fall rates could explain a significant proportion of the between-center variation in the incidence of limb fracture across Europe. PMID- 12531569 TI - Preparing for weapons of mass destruction. PMID- 12531568 TI - Cyclical behavior of bone remodeling and bone loss in healthy women after menopause: results of a prospective study. AB - Annual changes in lumbar bone mineral density (LBMD) and bone remodeling markers were measured in 238 healthy pre- and postmenopausal women, aged 45-74 years. The subjects were divided into groups according to their menstrual status and years since menopause. The results obtained indicate that bone loss is not a constant process over time but rather exhibits cyclical damping oscillations. When the log linear trend of LBMD decrement was transformed into a constant by considering annual percentage changes, the presence of a cyclical component of 7 years was evident. By employing a harmonic regression model, the cyclical component was also statistically significant on baseline data. The cyclical behavior of LBMD decrement corresponded to an analogous behavior of the bone remodeling markers. These results suggest that a lack of estrogen acts as a synchronizer on bone remodeling by triggering a latent cyclical rhythm of bone loss that persists throughout life after menopause. The existence of a chronobiological rhythm of bone loss starting after menopause, if confirmed, could have important clinical implications. PMID- 12531570 TI - In-situ lobular neoplasia: time for an awakening. PMID- 12531571 TI - New ways to stop biofilm infections. PMID- 12531572 TI - Diagnosing smallpox in possible bioterrorist attack. PMID- 12531573 TI - Is HIV superinfection worrisome? PMID- 12531574 TI - Fomite transmission with head lice: a continuing controversy. PMID- 12531576 TI - Adverse events associated with dietary supplements: an observational study. AB - BACKGROUND: Adverse events associated with dietary supplements are difficult to monitor in the USA, because such products are not registered before sale, and there is little information about their content and safety. METHODS: In 1998, 11 poison control centres in the USA recorded details of 2332 telephone calls about 1466 ingestions of dietary supplements, in 784 of which patients had symptoms. We used a multitiered review process (kappa 0.42) to select 489 cases for whom we were at least 50% certain that their negative events were associated with dietary supplements. We aimed to assess the effects of multiple ingredients and long-term use, and collated data for patterns of use and information resources. FINDINGS: A third of events were of greater than mild severity. We noted both new and previously reported associations that included myocardial infarction, liver failure, bleeding, seizures, and death. Increased symptom severity was associated with use of several ingredients, long-term use, and age. Paediatric exposures were more often unintentional than were adult ingestions, and treatment of disease was the reason for supplement use in at least 28% of reports. Most products and ingredients were not identified in the information database (Poisindex) used by poison control centres, and specific adverse events were reported variably among five additional sources. INTERPRETATION: Dietary supplements are associated with adverse events that include all levels of severity, organ systems, and age groups. Associations between adverse events and ingredients are difficult to verify if a product has more than one ingredient, and because of incomplete information systems. Research into hazards and risks of dietary supplements should be a priority. PMID- 12531577 TI - Analysis of pooled data from the randomised controlled trials of endarterectomy for symptomatic carotid stenosis. AB - BACKGROUND: Endarterectomy reduces risk of stroke in certain patients with recently symptomatic internal carotid stenosis. However, investigators have made different recommendations about the degree of stenosis above which surgery is effective, partly because of differences between trials in the methods of measurement of stenosis. To accurately assess the overall effect of surgery, and to increase power for secondary analyses, we pooled trial data and reassessed carotid angiograms. METHODS: We pooled data from the European Carotid Surgery Trial (ECST), North American Symptomatic Carotid Endarterectomy Trial, and Veterans Affairs trial 309 from the original electronic data files. Outcome events were re-defined, if necessary, to achieve comparability. Pre-randomisation carotid angiograms from ECST were re-measured by the method used in the other two trials. RESULTS: Risks of main outcomes in both treatment groups and effects of surgery did not differ between trials. Data for 6092 patients, with 35000 patient years of follow-up, were therefore pooled. Surgery increased the 5-year risk of ipsilateral ischaemic stroke in patients with less than 30% stenosis (n=1746, absolute risk reduction -2.2%, p=0.05), had no effect in patients with 30-49% stenosis (1429, 3.2%, p=0.6), was of marginal benefit in those with 50-69% stenosis (1549, 4.6%, p=0.04), and was highly beneficial in those with 70% stenosis or greater without near-occlusion (1095, 16.0%, p<0.001). There was a trend towards benefit from surgery in patients with near-occlusion at 2 years' follow-up (262, 5.6%, p=0.19), but no benefit at 5 years (-1.7%, p=0.9). INTERPRETATION: Re-analysis of the trials with the same measurements and definitions yielded highly consistent results. Surgery is of some benefit for patients with 50-69% symptomatic stenosis, and highly beneficial for those with 70% symptomatic stenosis or greater but without near-occlusion. Benefit in patients with carotid near-occlusion is marginal in the short-term and uncertain in the long-term. PMID- 12531578 TI - Combination treatment of angiotensin-II receptor blocker and angiotensin converting-enzyme inhibitor in non-diabetic renal disease (COOPERATE): a randomised controlled trial. AB - BACKGROUND: Present angiotensin-converting-enzyme inhibitor treatment fails to prevent progression of non-diabetic renal disease. We aimed to assess the efficacy and safety of combined treatment of angiotensin-converting-enzyme inhibitor and angiotensin-II receptor blocker, and monotherapy of each drug at its maximum dose, in patients with non-diabetic renal disease. METHODS: 336 patients with non-diabetic renal disease were enrolled from one renal outpatient department in Japan. After screening and an 18-week run-in period, 263 patients were randomly assigned angiotensin-II receptor blocker (losartan, 100 mg daily), angiotensin-converting-enzyme inhibitor (trandolapril, 3 mg daily), or a combination of both drugs at equivalent doses. Survival analysis was done to compare the effects of each regimen on the combined primary endpoint of time to doubling of serum creatinine concentration or end-stage renal disease. Analysis was by intention to treat. FINDINGS: Seven patients discontinued or were otherwise lost to follow-up. Ten (11%) of 85 patients on combination treatment reached the combined primary endpoint compared with 20 (23%) of 85 on trandolapril alone (hazard ratio 0.38, 95% CI 0.18-0.63, p=0.018) and 20 (23%) of 86 on losartan alone (0.40, 0.17-0.69, p=0.016). Covariates affecting renal survival were combination treatment (hazard ratio 0.38, 95% CI 0.18-0.63, p=0.011), age (1.30, 1.03-2.29, p=0.009), baseline renal function (1.80, 1.02 2.99, p=0.021), change in daily urinary protein excretion rate (0.58, 0.24-0.88, p=0.022), use of diuretics (0.80, 0.30-0.94, p=0.043), and antiproteinuric response to trandolapril (0.81, 0.21-0.91, p=0.039). Frequency of side-effects with combination treatment was the same as with trandolapril alone. INTERPRETATION: Combination treatment safely retards progression of non-diabetic renal disease compared with monotherapy. However, since some patients reached the combined primary endpoint on combined treatment, further strategies for complete management of progressive non-diabetic renal disease need to be researched. PMID- 12531579 TI - Atypical lobular hyperplasia as a unilateral predictor of breast cancer risk: a retrospective cohort study. AB - BACKGROUND: Clinical decisions about atypical lobular hyperplasia are based on the belief that later invasive breast-cancer risk is equal in both breasts. We aimed to show laterality and subsequent risk implications of invasive breast cancer in women with atypical lobular hyperplasia. METHODS: We did a retrospective cohort study of 252 women who had undergone 261 benign surgical biopsies that showed atypical lobular hyperplasia from 1950 to 1985, as part of the Nashville Breast Studies. Primary outcomes were development of invasive breast cancer and laterality of cancer compared with side of the biopsied breast. FINDINGS: 50 (20%) of 252 women treated by biopsy only developed invasive breast cancer. Relative risk of breast cancer in women with atypical lobular hyperplasia was 3.1 (95% CI 2.3-4.3, p<0.0001). Of these 50 women, the breast with invasive cancer was the same breast diagnosed with atypical lobular hyperplasia (ipsilateral) in 34 (68%) and the contralateral breast in 12 (24%). The ratio of ipsilateral/ contralateral cancers for atypical lobular hyperplasia without other atypical lesions was 17/5. For six women with atypical lobular hyperplasia plus atypical ductal hyperplasia, the ratio was 1/1. INTERPRETATION: Invasive carcinoma after atypical lobular hyperplasia is about three times more likely to arise in the breast diagnosed with atypical lobular hyperplasia than in the opposite breast without these initial findings. Our findings suggest a model of premalignancy for atypical lobular hyperplasia intermediate between a local precursor and a generalised risk for both breasts. See Commentary page 96 PMID- 12531580 TI - Stimulation of Staphylococcus epidermidis growth and biofilm formation by catecholamine inotropes. AB - BACKGROUND: Bacterial colonisation of indwelling medical devices by coagulase negative staphylococci is a prevalent risk in intensive-care units. Factors determining biofilm formation and progression to catheter- related infection are incompletely understood. We postulated that administration of inotropic agents via indwelling intravenous catheters may stimulate growth and formation of biofilms by Staphylococcus epidermidis. METHODS: Inocula representing physiologically relevant infecting doses of S epidermidis were incubated in a minimum medium supplemented with fresh human plasma in the presence or absence of pharmacological concentrations of norepinephrine or dobutamine. Biofilm formation on polystyrene and medical-grade silicone was examined. After incubation, cultures were assessed for growth and formation of biofilms by colony counting and scanning electronmicroscopy. The production of exopolysaccharide, a major constituent of S epidermidis biofilms, was also assessed by use of immunofluorescence microscopy. FINDINGS: Incubation of S epidermidis with catecholamine inotropes in the presence of human plasma resulted in a significant increase in growth compared with control on both polystyrene and silicone surfaces, with pronounced increases in biofilm formation as visualised by scanning electronmicroscopy. Experiments with transferrin labelled with radioactive iron showed the ability of catecholamine inotropes to facilitate acquisition of iron by S epidermidis. Immunofluorescence microscopy revealed extensive exopolysaccharide production associated with S epidermidis biofilms. INTERPRETATION: The ability of catecholamine inotropic drugs to stimulate bacterial proliferation and biofilm formation may be an aetiological factor in the development of intravascular catheter colonisation and catheter-related infection. The removal of iron from transferrin for subsequent use by S epidermidis is a possible mechanism by which catecholamine inotropes stimulate bacterial growth as biofilms. PMID- 12531581 TI - An unconscious girl with sickle-cell disease. PMID- 12531582 TI - Severe acute interstitial pneumonia and gefitinib. AB - Gefitinib is an oral selective inhibitor of the epidermal growth factor receptor tyrosine kinase that is an effective treatment for patients with advanced non small cell lung cancer who do not respond to platinum-based chemotherapy. We assessed four patients who had non-small cell lung cancer causing severe acute interstitial pneumonia in association with gefitinib. Although two patients recovered after treatment with steroids, the other two died from progressive respiratory dysfunction. On the basis of autopsies and bilateral distribution of diffuse ground-glass opacities in chest CTs, we diagnosed diffuse alveolar damage, which was consistent with acute interstitial pneumonia. Patients with interstitial pneumonia also had other pulmonary disorders such as previous thoracic irradiation and poor performance status. Physicians should be aware of the alveolar damage induced by gefitinib, especially for patients with these characteristic features. PMID- 12531583 TI - Intact fetal cells in maternal plasma: are they really there? AB - Rare fetal cells can be recovered from maternal blood, which suggests that non invasive prenatal diagnosis is possible. However, recovery and analysis of fetal cells from blood is complex, and sensitivity is low because of the rarity of these cells in the maternal circulation. An alternative strategy, which suggested that intact fetal cells can be found in maternal plasma by use of simple enrichment methods, has been reported. We aimed to replicate this technique. However, five independent laboratories were unable to identify any intact male cells from the plasma of 38 women known to be carrying male fetuses. Although apoptotic intact fetal cells could contribute to the detection of fetal DNA in maternal plasma, we believe that recovery of these cells is difficult and not clinically practical. PMID- 12531584 TI - Original biological reconstruction of the hip in a 4-year-old girl. AB - We resected the proximal femur in a 4-year-old child who had Ewing's sarcoma. We reconstructed her hip by an autotransplant of the ipsilateral proximal fibula with its vascular supply, inserted inside a massive bone allograft, and fixed to the residual femur by a plate. Serial postoperative imaging showed the progressive hypertrophy of the transplanted fibula, whose head displayed impressive tridimensional growth inside the acetabulum. 4 years after surgery, hip motility is 100 degrees in flexion, 10 degrees in extension, and 30 degrees in abduction, and the patient's limb is weight bearing. PMID- 12531585 TI - US human research committee draws fire. PMID- 12531586 TI - Dutch elections mark testing time for health of the nation. PMID- 12531588 TI - Exposure to oil spill has detrimental effect on clean-up workers' health. PMID- 12531589 TI - Polio resurgence in Indian state causes concern. PMID- 12531592 TI - Bill Frist takes the helm of the US Senate. PMID- 12531593 TI - Atopic dermatitis. AB - Atopic dermatitis is a highly pruritic chronic inflammatory skin disorder affecting 10-20% of children worldwide. Symptoms can persist or begin in adulthood. It is also the most common cause of occupational skin disease in adults. This disease results from an interaction between susceptibility genes, the host's environment, pharmacological abnormalities, skin barrier defects, and immunological factors. New management approaches have evolved from advances in our understanding of the pathobiology of this common skin disorder. PMID- 12531594 TI - Laboratory diagnosis. PMID- 12531595 TI - Transfusion medicine: looking to the future. AB - The evolution of transfusion medicine into a clinically oriented discipline emphasising patient care has been accompanied by challenges that need to be faced as specialists look to the future. Emerging issues that affect blood safety and blood supply, such as pathogen inactivation and more stringent donor screening questions, bring new pressures on the availability of an affordable blood supply. Imminent alternatives for management of anaemia, such as oxygen carriers, hold great promise but, if available, will require close oversight. With current estimates of HIV or hepatitis C viral (HCV) transmission approaching one in 2000000 units transfused, keeping to a minimum bacterial contamination of platelet products (one in 2000) and errors in transfusion, with its estimated one in 800000 mortality rate, assume great urgency. Finally, serious difficulties in blood safety and availability for poor, developing countries require innovative strategies and commitment of resources. PMID- 12531596 TI - WHO and the challenges of the next decade. PMID- 12531597 TI - Addressing inequalities: the role for the new WHO Director General. PMID- 12531598 TI - Two key issues for the new WHO leadership. PMID- 12531599 TI - Breastfeeding, atopy, and asthma. PMID- 12531600 TI - Breastfeeding, atopy, and asthma. PMID- 12531601 TI - Breastfeeding, atopy, and asthma. PMID- 12531603 TI - Breast cancer and breastfeeding. PMID- 12531604 TI - Breast cancer and breastfeeding. PMID- 12531605 TI - Tamoxifen for breast-cancer prevention. PMID- 12531606 TI - Tamoxifen for breast-cancer prevention. PMID- 12531609 TI - Genetic testing for BRCA1 mutation in the UK. PMID- 12531611 TI - Risk of ovarian cancer in breast-cancer patients with a family history of either. PMID- 12531613 TI - What is a near miss? PMID- 12531614 TI - Pancreatic cancer and basal-cell carcinoma. PMID- 12531615 TI - Debate over Japanese renaming of term for schizophrenia. PMID- 12531616 TI - Dengue fever and dengue haemorrhagic fever. PMID- 12531617 TI - Deferiprone versus desferrioxamine in thalassaemia, and T2* validation and utility. PMID- 12531618 TI - Deferiprone versus desferrioxamine in thalassaemia, and T2* validation and utility. PMID- 12531619 TI - Deferiprone versus desferrioxamine in thalassaemia, and T2* validation and utility. PMID- 12531621 TI - Deferiprone versus desferrioxamine in thalassaemia, and T2* validation and utility. PMID- 12531622 TI - WHO's next Director General. PMID- 12531629 TI - Immune responses elicited against multiple enterotoxigenic Escherichia coli fimbriae and mutant LT expressed in attenuated Shigella vaccine strains. AB - Shigella and enterotoxigenic Escherichia coli (ETEC) continue to be important causes of diarrheal disease in infants and young children in developing countries and are major etiologic agents of traveler's diarrhea. Since attenuated strains of Shigella have been developed as live oral vaccines against shigellosis, we have adapted these attenuated Shigella strains to serve as carriers of ETEC antigens, thereby constituting a hybrid vaccine. Since protective immunity against ETEC is largely directed against fimbrial antigens (of which there are multiple antigenic types), we have individually expressed four different ETEC fimbriae, including CFA/I, CS2, CS3, and CS4, using deltaguaBA attenuated Shigella vaccine strain CVD 1204 as a prototype live vector. Following mucosal (intranasal) immunization of guinea pigs, serum IgG and mucosal IgA responses were elicited against each fimbrial type. An additional strain was constructed expressing a detoxified version of the human ETEC variant of heat labile toxin (LThK63). Following mucosal immunization of guinea pigs with a mixed inoculum containing five Shigella strains each expressing a different ETEC antigen, immune responses were observed against each ETEC antigen plus the Shigella vector. PMID- 12531630 TI - Oral immunization of adult volunteers with microencapsulated enterotoxigenic Escherichia coli (ETEC) CS6 antigen. AB - As a step in the development of an oral vaccine against ETEC, we evaluated the safety and immunogenicity of CS6, a polymeric protein commonly found on the surface of ETEC. Formulations included 1 and 5mg doses of CS6, either encapsulated in biodegradable polymer poly(D, L)-lactide-co-glycolide (PLG), or as free protein, administered orally in a solution of either normal saline or a rice-based buffer. Three doses of CS6 were given at 2-week intervals. Blood was collected immediately before and 7 days after each dose. All formulations were well tolerated. Four of five volunteers who received 1mg CS6 in PLG microspheres with buffer had significant IgA ASC responses (median=30 ASC per 10(6) PBMC) and significant serum IgG responses (median=3.5-fold increase). Oral administration of these prototype ETEC vaccine formulations are safe and can elicit immune responses. The ASC, serum IgA, and serum IgG responses to CS6 are similar in magnitude to the responses after challenge with wild-type ETEC [Coster et al., unpublished data]. Further studies are underway to determine whether these immune responses are sufficient for protection. PMID- 12531631 TI - Prospects for a mucosally-administered vaccine against Helicobacter pylori. PMID- 12531632 TI - Anticipating rotavirus vaccines: epidemiology and surveillance of rotavirus in South Africa. AB - Rotavirus infection is associated with acute infantile gastroenteritis in infants and young children globally. In South Africa, rotavirus infection has been shown to be associated with approximately one-quarter of all diarrhoeal admissions to hospital. Rotavirus infection predominantly occurs in infants less than 12 months of age (75%) and has a peak of shedding during the cooler, drier months of the year. A secondary peak during the spring has been observed. Multiple infections with rotavirus and at least one other microbial agent are common. The circulating VP7 serotypes and VP4 genotypes have been determined in various regions of South Africa and show a geographic specific distribution. A decade previously, P[8]G1 or G4 strains predominated, and P[4]G2 strains occurred in an epidemic pattern in one region. More recently, rotavirus strains with P[6] genotype have become common and novel VP7/VP4 genotype combinations are occurring across the country. G9 strains have been reported from Cape Town to Vendaland. The circulating rotavirus types observed in this study add to the knowledge of the natural history of rotavirus infection and provide the groundwork to consider future vaccine strategies. PMID- 12531633 TI - Rotavirus strains circulating in Africa during 1996-1999: emergence of G9 strains and P[6] strains. AB - Rotavirus infection is associated with 150000-200000 deaths annually in Africa. Although the withdrawal of the RotaShield vaccine has been a major setback in rotavirus vaccine development, new vaccine candidates are under development and approaching phase II and III trials. Before these trials could be conducted in Africa, a comprehensive survey of the circulating VP7 serotypes and VP4 genotypes is required. During the past 3 years, over 3000 rotavirus-positive specimens from several African countries have been analysed. RT-PCR techniques for the VP7 and VP4 genotypes and by monoclonal antibodies to the VP6 subgroup and VP7 serotype have been performed. Almost 75% of the strains were typed by the VP7 monoclonal antibodies or RT-PCR. VP4 genotyping was done in approximately half of these strains. The predominant strains circulating across Africa during 1996-1999 were P[6]G1 and P[6]G3 strains. Geographic differences were noted and West Africa displayed the most diverse strains with G3/8 and G1/3 "mosaic" viruses occurring commonly. G9 strains were identified in several countries indicating that the strain is emerging in Africa too. G9 was the predominant strain in certain countries during 1999. The circulating types observed will have implications for the new rotavirus vaccine candidates. PMID- 12531634 TI - Immunogenicity and protective efficacy of a formalin-inactivated rotavirus vaccine combined with lipid adjuvants. AB - The immunogenicity and protective efficacy of inactivated rotavirus vaccine administered intramuscularly with lipid adjuvants; MPL (monophosphoryl lipid A from Salmonella minnesota) or L3 (monooleate/lauric acid) was evaluated in an infant mouse model. Purified and formalin-inactivated rhesus rotavirus (I-RRV) combined with one of the adjuvants were administered to female balb/c mice at 0, 4 and 8 weeks. High serum IgG antibody titers developed in all vaccinated groups; I-RRV (GMT 45524+/-9819), I-RRV-MPL (GMT 190637+/-64250) and I-RRV-L3 (GMT 126266+/-27553). The formalin-inactivation procedure preserved neutralizing epitopes and elicited high neutralizing antibody titers; I-RRV (GMT 43053 S.E.M.+/-4189), I-RRV-MPL (GMT 66398 S.E.M.+/-20202) and I-RRV-L3 (GMT 60887 S.E.M.+/-10750). All offsprings to immunized dams were protected against clinical diarrhea upon oral challenge with RRV. The IgG1/IgG2a ratio was in all immunized groups approximately 1 suggesting development of a balanced Th1/Th2 response. PMID- 12531635 TI - A modified cholera holotoxin CT-E29H enhances systemic and mucosal immune responses to recombinant Norwalk virus-virus like particle vaccine. AB - In this study, we evaluated the potential of a genetically modified cholera toxin, CT-E29H as an adjuvant for recombinant Norwalk virus like particle (NV VLP) vaccine. This detoxified mutant, containing E to H substitution at amino acid 29 of the CT-A1 subunit, was administered with a recombinant Norwalk virus like particle vaccine to Balb/c mice by mucosal routes to monitor the induction of mucosal, humoral and cellular responses. We observed that a low dose of NV-VLP (5 microg) with the adjuvant delivered by the intranasal route (IN) was more effective than the highest dose (200 microg) delivered by oral route at inducing both cellular and NV-VLP specific IgG and IgA responses. Higher counts of antigen specific IgA secreting cells were observed in the Peyer's Patches (PP) following delivery of the vaccine with CT-E29H as compared to delivery of vaccine by mucosal routes without CT-E29H. Furthermore, there was an increase in antigen specific cells producing IL-4 from animals that received the vaccine with the adjuvant. Delivery of the vaccine by the oral route results in antigen specific CD4(+) and CD8(+) T cells in PP and spleen. Addition of CT-E29H results in an increase of antigen specific CD4(+) cell population in PP and both CD4(+) and CD8(+) populations in the spleen. These cellular and cytokine responses suggest that combining the vaccine with CT-E29H results in a stronger Th2 type response. Collectively, these results indicate that immune responses to NV-VLP vaccine are qualitatively and quantitatively improved when the vaccine is delivered along with CT-E29H, and thus merits its further consideration as a mucosal adjuvant. PMID- 12531636 TI - Construction of artificial virus-like particles exposing HIV epitopes, and the study of their immunogenic properties. AB - One of the major problems in the development of successful recombinant vaccines against human immunodeficiency virus (HIV) is that of correct identification of a safe and effective vaccine delivery system with which to induce protective immunity using soluble protein antigens. An original method for constructing artificial immunogens in the form of spherical particles with yeast dsRNA in the center and hybrid proteins exposing epitopes of an infectious agent on the surface is reported. The dsRNA and the proteins were linked with spermidine polyglucin-glutathione conjugates. Particles exposing HIV-1 epitopes were constructed, and their immunogenicity tested. PMID- 12531637 TI - The information network of senior citizens in Geneva, Switzerland, and progress in flu vaccination coverage between 1991 and 2000. AB - Switzerland has lagged behind other industrialized countries in increasing vaccination coverage against flu in the elderly population. The information campaign "United against Flu", started in Geneva in 1993, gradually extended to other French and Italian speaking cantons in Switzerland and indirectly affected German-speaking cantons. Activities developed include the production of TV spots, press conferences, information forwarded to health professionals, an Internet site and information material such as leaflets and posters to risk groups. The campaign is evaluated by repeated surveys that measure vaccination coverage as well as network of informants, knowledge and perceptions in the geriatric population. Vaccination coverage of the geriatric population in Geneva canton has increased from 29% in 1991 to 59% in the year 2000. PMID- 12531638 TI - Immigration flows and new epidemiological evidence in southern Italy. AB - Since 1991, immigration flows have had a relevant impact on the area of Salento, in southern Italy, with thousands of landings every year. For this reason, both temporary and permanent shelters were created. At these centres a team of doctors works permanently and full-time, carefully evaluating the immigrants' state of health and performing the sanitary interventions needed and/or provided by the law. Recently, 55 chickenpox cases were reported among Sri Lankan immigrants, four cases among subjects already sheltered in the centres who became infected after the arrival of the Sri Lankan group, and one in an Italian child. The Sri Lankan immigrants affected by chickenpox were aged 18-41 years and all recovered without sequelae. A seroepidemiological study was also performed. This was the first chickenpox epidemic ever occurred in a shelter for immigrants in Italy. This should induce attention towards the new and unpredictable sanitary emergencies that may occur in such contexts. PMID- 12531639 TI - Animal models paving the way for clinical trials of attenuated Salmonella enterica serovar Typhi live oral vaccines and live vectors. AB - Attenuated Salmonella enterica serovar Typhi (S. Typhi) strains can serve as safe and effective oral vaccines to prevent typhoid fever and as live vectors to deliver foreign antigens to the immune system, either by the bacteria expressing antigens through prokaryotic expression plasmids or by delivering foreign genes carried on eukaryotic expression systems (DNA vaccines). The practical utility of such live vector vaccines relies on achieving a proper balance between minimizing the vaccine's reactogenicity and maximizing its immunogenicity. To advance to clinical trials, vaccine candidates need to be pre-clinically evaluated in relevant animal models that attempt to predict what their safety and immunogenicity profile will be when administered to humans. Since S. Typhi is a human-restricted pathogen, a major obstacle that has impeded the progress of vaccine development has been the shortcomings of the animal models available to assess vaccine candidates. In this review, we summarize the usefulness of animal models in the assessment of the degree of attenuation and immunogenicity of novel attenuated S. Typhi strains as vaccine candidates for the prevention of typhoid fever and as live vectors in humans. PMID- 12531640 TI - Memory T cells and vaccines. AB - T lymphocytes play a central role in the generation of a protective immune response in many microbial infections. After immunization, dendritic cells take up microbial antigens and traffic to draining lymph nodes where they present processed antigens to naive T cells. These naive T cells are stimulated to proliferate and differentiate into effector and memory T cells. Activated, effector and memory T cells provide B cell help in the lymph nodes and traffic to sites of infection where they secrete anti-microbial cytokines and kill infected cells. At least two types of memory cells have been defined in humans based on their functional and migratory properties. T central-memory (T(CM)) cells are found predominantly in lymphoid organs and can not be immediately activated, whereas T effector-memory (T(EM)) cells are found predominantly in peripheral tissue and sites of inflammation and exhibit rapid effector function. Most currently licensed vaccines induce antibody responses capable of mediating long term protection against lytic viruses such as influenza and small pox. In contrast, vaccines against chronic pathogens that require cell-mediated immune responses to control, such as malaria, Mycobacterium tuberculosis (TB), human immunodeficiency virus (HIV) and hepatitis C virus (HCV), are currently not available or are ineffective. Understanding the mechanisms by which long-lived cellular immune responses are generated following vaccination should facilitate the development of safe and effective vaccines against these emerging diseases. Here, we review the current literature with respect to memory T cells and their implications to vaccine development. PMID- 12531641 TI - The Echinococcus multilocularis 14-3-3 protein protects mice against primary but not secondary alveolar echinococcosis. AB - Alveolar echinococcosis (AE), caused by the larval stage (metacestode) of the tapeworm Echinococcus multilocularis, exhibits very similar disease characteristics in humans and rodents. Recently, it has been shown that an over expression of the parasite 14-3-3 protein could be associated to the proliferative growth of the E. multilocularis metacestode. We now demonstrate the expression of this protein at the E. multilocularis oncospheral stage as well. A recombinant E. multilocularis 14-3-3 protein (E14t) was used to vaccinate mice against either primary or secondary experimental E. multilocularis infection in BALB/c mice. Conversely to non-vaccinated but control infected mice, which developed a very weak anti-E14t response during infection, the response elicited in the E14t-vaccinated and subsequently infected animals exhibited a strong reactivity against the parasite 14-3-3 protein. Major differences became apparent between secondarily and primarily infected animals: whereas no protection against secondary infection was achieved by vaccination, vaccinated animals were protected by 97% against challenge primary infection with 2000 E. multilocularis eggs. Consequently, the parasite 14-3-3 molecule appears crucially involved in the early stage of the host-parasite interplay and exhibits potential to be used as target molecule for the development of protective tools against AE. PMID- 12531642 TI - Measles surveillance and control in Tunisia: 1979-2000. AB - BACKGROUND: The aim of this study was to analyze measles epidemiology in Tunisia after the introduction of specific vaccine in 1979 and the results of the serological investigation of suspected cases, introduced as part of the National Program for Measles Elimination recently implemented. METHODS: Available data were used to examine trends in measles epidemiology from 1979 to 2000: number of reported cases, patient's age, reporting date, epidemiological link with similar cases and laboratory confirmation. Serological investigation included the detection, by ELISA, of measles and rubella IgMs in 542 suspected measles cases sampled from 1997 to 2000. RESULTS: Measles coverage level increased gradually and was maintained to over 90% since 1992. In parallel, the annual incidence of reported cases declined and outbreaks became less frequent, the latest occurred in 1992. Measles-specific IgMs were detected in only nine patients who received measles vaccine few days before blood collection, anti-rubella IgMs were detected in 52% of cases. CONCLUSION: Vaccination strategies including routine and supplemental immunizations, implemented in Tunisia, achieved a substantial decrease in measles incidence. Virological results highlight frequent confusion, at the clinical level, with the other etiologies of rash and fever and the importance of systematic serological confirmation of cases. PMID- 12531643 TI - The analysis of immune responses of a novel CC-chemokine gene from Japanese flounder Paralichthys olivaceus. AB - A novel CC-chemokine gene was isolated from the Japanese flounder Paralichthys olivaceus by expressed sequence tag analysis. The function of this CC-chemokine gene was studied by DNA injection. To investigate the immune responses to the CC chemokine, a plasmid construct containing the novel CC-chemokine and a CMV promoter was injected into the epaxial muscle of Japanese flounder. Quantification of CC-chemokine protein expressed in serum on 1, 3 and 5 days after plasmid injection were estimated by ELISA. CC-chemokine gene injection increased the migration of phagocytic cells. Macrophage functions such as production of superoxide anion and phagocytosis were also stimulated by this gene injection. Thus, this gene from Japanese flounder has functional similarities to that of a mammalian CC-chemokine gene. PMID- 12531644 TI - B- and T-cell responses to the mycobacterium surface antigen PstS-1 in the respiratory tract and adjacent tissues. Role of adjuvants and routes of immunization. AB - Induction of mucosal immunity in the respiratory tract is crucial for protection against respiratory infections. Here, we have investigated the effects of the routes of immunization as well as of three different adjuvants on the induction of mucosal immune responses. Mice were immunized using intranasal (i.n.) or intraperitoneal (i.p.) routes with the mycobacterium PstS-1 antigen. Cholera toxin (CT), detoxified pertussis toxin (detPT) and RU 41.740 from Klebsiella pneumoniae were compared as mucosal adjuvants. Our data showed that i.n. route of immunization induced the most favorable stimulation of mucosal antigen-specific IgA responses supported by mixed Th cells producing IL-4, IL-5, IFN-gamma. In contrast, i.p. immunizations elicited only enhancement of systemic responses, predominantly of the Th2 type. Furthermore, the use of CT as mucosal adjuvant resulted in the stimulation of a mixed Th cell response whereas detPT evoked mainly Th2 type of responses. Likewise CT, the RU 41.740 adjuvant elicited a mixed Th cell response, albeit supported by much lower numbers of CD4(+) T-cells. Thus, i.n. route of immunization favors the induction of mucosal and systemic immune responses, while the Th cell development at mucosal inductive site is influenced by the adjuvant used for immunizations. PMID- 12531645 TI - When can a clinical trial be called 'randomized'? AB - It is widely recognized that, in the context of the evaluation of medical interventions, randomized clinical trials constitute the gold standard. This is because randomization tends to balance both measured and unmeasured baseline characteristics, allows for masking, and provides a basis for inference. It is understandable, then, that investigators would wish to utilize this methodology whenever it is feasible to do so. Unfortunately, some studies are labeled as randomized when in fact they are not. These studies then receive more credibility, and influence medical practice, more than they deserve to. After reviewing the benefits of randomization, paying particular attention to the specific aspects of randomization that confer each benefit, we will explore the issue of what constitutes a randomized study. PMID- 12531646 TI - Measles vaccine efficacy during an epidemic in 1998 in the highly vaccinated population of Poland. AB - A measles epidemic with 2255 reported cases occurred in Poland between November 1997 and July 1998, despite high vaccination coverage since the 1980s. Cases occurred at all ages less than 30 years but showed two distinct peaks: young, unvaccinated children born in 1996-1997 and once vaccinated young adults born 1976-1982. The 60% of cases were among persons aged 15 years or more. A cohort study was used to investigate measles vaccine efficacy. The efficacy of a single dose of vaccine exceeded 90% and the efficacy of two doses exceeded 99% in all age-groups. These results demonstrate both the high efficacy of the monovalent measles vaccines used in Poland and the benefit of a second dose of vaccine. PMID- 12531647 TI - Difficulties in measles elimination: prevalence of measles antibodies before and after mass vaccination campaign in Laos. AB - Responding to the declaration of regional polio eradication in 2000, Western Pacific Region (WPR) has been strengthening the efforts for measles elimination. Nevertheless, we questioned the feasibility of the measles elimination in Laos, where measles was still endemic. This study was undertaken with the aim to examine the impact of mass measles vaccination campaign, by comparing the prevalence of measles antibodies between before and after the campaign in 2000, among the children aged 9 months to 4 years in the two pilot sites. The constraints of mass vaccination campaign have been explored in this study. It was also suggested that more efforts should be put on reconstruction of the existing routine immunization service in Laos, which had been weakened by aggressive eradication programme. PMID- 12531648 TI - The binding of CpG-oligodeoxynucleotides to cell-surface and its immunostimulatory activity are modulated by extracellular acidic pH. AB - Both the binding of CpG-oligodeoxynucleotides (CpG-ODNs) to cell-surface and its immunostimulatory activity were modulated by extracellular pH in present study. At neutral pH (pH 7.4), the binding of CpG-ODN to splenocyte-surface, as well as that of non-CpG-ODN, was competitively inhibited by non-specific DNA-Herring sperm DNA in a dose dependent manner, indicating their binding sites have no specificity for CpG-motif. When the extracellular pH shifted to acidic (pH 6.4), however, their binding to cell-surface markedly increased, and only the binding of non-CpG-ODN instead of CpG-ODN was inhibited by Herring sperm DNA, implying such pH change enabled CpG-ODN bind to its specific binding-site. Consistently, lymphocytes appeared more sensitive to the stimulation of CpG-ODN at acidic pH, and Herring sperm DNA inhibited the CpG-ODN-induced TNF production from splenocytes at pH 7.4, but not at pH 6.4. These results suggest the existence of membrane receptor that specifically engages CpG-ODN with high affinity only at acidic pH, and support the hypothesis that the binding CpG-ODN to its specific membrane receptor and subsequently triggering of CpG-related signaling occurred within acidified endosomes. PMID- 12531649 TI - Response of ELA-A1 horses immunized with lipopeptide containing an equine infectious anemia virus ELA-A1-restricted CTL epitope to virus challenge. AB - Lipopeptide containing an ELA-A1-restricted cytotoxic T lymphocyte (CTL) epitope from the envelope surface unit (SU) protein of the EIAV(WSU5) strain was used to immunize three horses having the ELA-A1 haplotype. Peptide-specific ELA-A1 restricted CTL were induced in all three horses, although these were present transiently in PBMC. These horses were further immunized with lipopeptide containing the corresponding CTL epitope from the EIAV(PV) strain. Then, the three immunized horses and three non-immunized horses were challenged by intravenous inoculation with 300 TCID(50) EIAV(PV). All horses developed cell free viremia, fever and thrombocytopenia. However, there was a statistically lower fever and thrombocytopenia severity score in the immunized group. Shorter duration of plasma viral load in two of the three immunized horses likely explains the less severe clinical disease in this group. Results indicate that lipopeptide immunization had a protective effect against development of clinical disease following virus challenge. PMID- 12531650 TI - DNA vaccination with plasmids containing various fragments of large segment genome of infectious bursal disease virus. AB - The present study was undertaken to determine the effectiveness in including VP2 gene of the large segment genome of infectious bursal disease virus (IBDV) in the DNA vaccine for protection of chickens against infectious bursal disease (IBD). Different fragments of the large segment gene of IBDV standard challenge (STC) strain were successfully amplified by reverse transcription-polymerase chain reaction (RT-PCR) followed by cloning into a eukaryotic expression plasmid vector (pCR3.1) as DNA vaccines. Transient expression of the encoded genes from various constructed plasmids was characterized in COS-7 cells by positive immunofluorescent staining with polyclonal or monoclonal antibody to IBDV. Chickens (1-day old) were intramuscularly injected with the individual plasmid three times at weekly intervals, challenged with IBDV strain STC at 21-day old, and observed for 10 days. Chickens receiving the plasmids containing VP2 genes, including VP243, VP24, or VP2 fragment, did not have clinical signs, mortality, and bursal atrophy and were effectively protected against IBDV infection. On the contrary, chickens receiving plasmids without containing VP2 genes, including VP4, VP3, or VP43 fragment, had marked bursal atrophy and were not protected against IBD. Antigen detection was correlated with protection; chickens protected from IBDV infection had undetectable IBDV antigen in bursae. Enzyme-linked immunosorbent assay (ELISA) antibody titer to IBDV was low or undetectable prior to or after challenge with IBDV in protected chickens receiving the plasmids containing the VP2 gene. The results indicate that inclusion of VP2 gene in the plasmid DNA is essential in achieving effective protection mediated by DNA vaccination against IBDV infection in chickens. PMID- 12531651 TI - A mucosally administered recombinant fusion protein vaccine against schistosomiasis protecting against immunopathology and infection. AB - We have constructed and efficiently produced and purified a candidate vaccine against schistosomiasis consisting of a novel hybrid protein in which two dominant T- and B-cell epitopes from Schistosoma mansoni 28 kDa glutathione-S transferase (Sm28GST) antigen (a.a 24-43 and 191-212) are fused to cholera toxin B subunit (CTB). Intranasal treatment of S. mansoni-infected mice with the hybrid protein, which similar to native CTB was assembled into receptor binding pentamers, significantly reduced total worm burden and liver egg counts due to the induction of Sm28GST-specific antibodies. Immunopathologic granuloma formation in the liver was also significantly suppressed and there was an almost complete suppression of delayed-type hypersensitivity reactions to both Sm28GST and to total soluble egg antigen in infected animals. The results suggest that this type of hybrid protein could be used as a combined anti-immunopathology and anti-infection vaccine against schistosomiasis. PMID- 12531652 TI - Comparison of the antitumor efficacies of Her-2/neu DNA vaccines inducing contrasting IgG immunity but comparable CTL activity in mice. AB - The relative importance of CTL and antibodies in rejecting Her-2/neu-expressing tumors was evaluated in preventive and therapeutic models by DNA vaccination. Four human Her-2/neu-expressing plasmids (pNeu(TM), pNeu(ECD), pNeu(TM-gDs), and pNeu(ECD-gDs)) were generated encoding either the transmembrane and extracellular domains or the extracellular domain. Interestingly, these plasmids demonstrated substantial difference in inducing Her-2/neu-specific serum IgG according to their signal sequence when injected in BALB/c mice. pNeu(TM) and pNeu(ECD) induced high serum IgG titers. pNeu(TM-gDs) and pNeu(ECD-gDs) induced low or very low serum IgG titers, respectively. As a result, mice vaccinated with not only pNeu(ECD) but also pNeu(ECD-gDs) exhibited complete eradication of a small number of tumor cells. Nevertheless, when the number of tumor cells was increased in a therapeutic model, only pNeu(ECD) exhibited statistically significant antitumor immunity. These studies demonstrate that strong CTL may be sufficient in tumor prevention, but the collaboration of CTL and antibody may be required in tumor therapy. PMID- 12531653 TI - Vaccine efficacy of the attenuated Erysipelothrix rhusiopathiae YS-19 expressing a recombinant protein of Mycoplasma hyopneumoniae P97 adhesin against mycoplasmal pneumonia of swine. AB - The attenuated Erysipelothrix rhusiopathiae YS-19 strain was constructed for the purpose of delivering the C-terminal portion of the Mycoplasma hyopneumoniae P97 adhesin to the mucosal surface of the respiratory tract of pigs. In this study, the efficacy of the YS-19 vaccine against mycoplasmal pneumonia of swine was evaluated. Animal experiments revealed that intranasal immunization of pigs with the YS-19 strain significantly reduced the severity of pneumonic lung lesions caused by M. hyopneumoniae infection. In YS-19-immunized pigs, P97-specific serum antibodies were not detected. However, when stimulated with the P97 protein, peripheral blood mononuclear cells from the YS-19-immunized pigs had a significantly higher stimulation index (P<0.05) than that of cells from control pigs at 7 days post-challenge. PMID- 12531654 TI - Salmonella typhi and S typhimurium derivatives harbouring deletions in aromatic biosynthesis and Salmonella Pathogenicity Island-2 (SPI-2) genes as vaccines and vectors. AB - The S. typhimurium strain (TML deltaaroC deltassaV) WT05, harbouring defined deletions in genes involved in both the aromatic biosynthesis pathway (aroC) and the Salmonella Pathogenicity Island-2 (SPI-2) (ssaV) was shown to be significantly attenuated in C57 BL/6 interferon gamma knockout mice following oral inoculation. Similarly, the S. typhi strain (Ty2 deltaaroC deltassaV) ZH9 harbouring the aroC and ssaV mutations propagated less efficiently than wild type in human macrophages. These studies demonstrated the attractive safety profile of the aroC ssaV mutant combination. Strains S. typhimurium (TML deltaaroC deltassaV ) WT05 and S. typhi (Ty2 deltaaroC deltassaV) ZH9 were subsequently tested as vaccine vectors to deliver E. coli heat-labile toxin (LT-B) mucosally to mice. Mice inoculated orally with S. typhimurium (TML deltaaroC deltassaV) WT05 expressing LT-B (WT05/LT-B) elicited high titres of both LT-specific serum IgG and intestinal IgA, although no specific IgA was detected in the vagina. Similarly, intranasal inoculation of mice with S. typhi (Ty2 deltaaroC deltassaV) ZH9 expressing LT-B (ZH9/LT-B) elicited even higher titres of LT-specific serum antibody as well as LT-specific Ig in the vagina. We conclude that deltaaroC deltassaV strains of Salmonella are highly attenuated and are promising candidates both as human typhoid vaccines and as vaccine vectors for the delivery of heterologous antigens. PMID- 12531655 TI - Suppression of allergen reactive Th2 mediated responses and pulmonary eosinophilia by intranasal administration of an immunodominant peptide is linked to IL-10 production. AB - The potential to induce systemic tolerance following exposure of the airway mucosa to soluble antigen, may be applied therapeutically for the treatment of allergic disease. Since the use of allergen can trigger IgE mediated inflammation, we investigated whether mucosal delivery of a peptide, containing an immunodominant epitope of the Der p1 allergen of house dust mite, can lead to CD4(+) Th2 cell tolerance and thus protect against airway inflammatory responses to inhalant allergen. The administration of microencapsulated peptide to the nasal mucosa of mice, protected against airway inflammation, with significant reductions in eosinophil infiltration into the airways following allergen challenge. Der p1 specific antibody levels in sera were not modulated. Allergen reactive CD4(+) T cells expressed a tolerized phenotype, with reduction in levels of the cytokines, IL-5, IL-13 and IFN-gamma although IL-10 levels were increased. The mucosal administration of a peptide containing an immunodominant region of an allergen can protect against the induction of systemic and local inflammatory responses to allergen challenge. PMID- 12531656 TI - B cell responses in gastric antrum and duodenum following oral inactivated Helicobacter pylori whole cell (HWC) vaccine and LT(R192G) in H pylori seronegative individuals. AB - To investigate whether B cell-specific responses could be elicited in the gastric mucosa of Helicobacter pylori (HP) naive subjects, five volunteers ingested three doses of a HP killed whole cell (HWC) vaccine with 25 microg of recombinant heat labile toxin (LT(R192G)). Two of three subjects had detectable LT(R192G) and HWC IgA antibody secreting cell (ASC) gastric responses. LT(R192G) and HWC responses in duodenal were 5-14-fold higher than those detected in antral biopsies (P<0.01 and P=0.05, respectively). These results provide the first evidence that specific gastric B cell responses can be induced in HP-non-infected individuals following oral immunization. PMID- 12531658 TI - Treatment of fractures of the fingers. What's new? PMID- 12531657 TI - Therapeutic efficacy of an E coli strain carrying an ovalbumin allergenic peptide as a fused protein to OMPC in a murine model of allergic airway inflammation. AB - An Escherichia coli strain expressing the ovalbumin (OVA) 323-329 allergenic peptide on the bacterial surface was evaluated for its ability to reduce the lung inflammatory response in mice allergic to OVA. BALB/c mice were rendered allergic by means of two intraperitoneal injections of OVA suspended in alum 5 days apart, and one intratracheal boost 1 week later. The mice were then treated with two intranasal, 1 week apart, doses of 4x10(9) E. coli-UH302 transformed with plasmids pST13 or pST13-OVA(323-339), which bear the OmpC porin from Salmonella enterica serovar Typhi or the OmpC with the OVA allergenic 323-339 amino acid sequence inserted in the external loop 5. The allergic inflammatory reaction was evaluated on day 31, finding that mice treated with E. coli-UH302-pST13-OVA reduced four to seven times perivascular and peribronchial infiltrates, mucus production, goblet cell hyperplasia and eosinophils when compared with mice treated with E. coli-UH302-pST13 or saline solution. These results were consistent with a significant decrease of IL-5 mRNA and induction of IFN-gamma mRNA in cells from bronchio-alveolar lavages (BAL). Specific serum IgE anti-OVA was also reduced, although the decrease did not reach statistical significance. These results demonstrate that the bacterial live vector bearing an allergenic peptide successfully moderated two important components of allergy, pulmonary inflammation and mucus overproduction. PMID- 12531659 TI - A prospective randomized controlled study of fixation of long oblique and spiral shaft fractures of the proximal phalanx: closed reduction and percutaneous Kirschner wiring versus open reduction and lag screw fixation. AB - Patients with an isolated spiral or long oblique fracture of the proximal phalanx were randomized into two groups. One was treated by closed reduction and Kirschner wire fixation and the second treated by open reduction and lag screw fixation. An independent observer assessed function, pain, movement, grip strength and intrinsic muscle function. X-rays were assessed for malunion. Thirty two patients were entered the study and 15 in the Kirschner wire and 13 in the lag screw group were reviewed at a mean follow-up of 40 months. There was no significant difference in the functional recovery rates or in the pain scores for the two groups. X-rays showed similar rates of malunion and there were no statistically significant differences in range of movement or grip strength. PMID- 12531660 TI - Avulsion fractures from the base of the proximal phalanges of the fingers. AB - Thirty-three patients with avulsion fractures from the base of the proximal phalanges were treated during a 6-year period. All eight fractures treated conservatively failed to unite and subsequently required surgery. The remaining 25 patients were treated with primary internal fixation using a single lag screw through a palmar approach. Surgery gave excellent results in all cases and all patients achieved a full range of movement within 3 weeks. PMID- 12531662 TI - Irreducible tuft fractures of the distal phalanx. AB - Two tuft fractures of the distal phalanx had a unique fracture pattern with significant displacement of the proximal fracture fragment into the pulp of the finger. Both fractures were irreducible by closed means, probably due to entrapment of bony spikes within the fibrous septa of the pulp or overlapping of the proximal and distal fracture fragments. PMID- 12531661 TI - The Ishiguro extension block technique for the treatment of mallet finger fracture: indications and clinical results. AB - The results of extension block Kirschner wire fixation for the treatment of mallet fractures of the distal phalanx were retrospectively assessed in 65 consecutive patients. The indications for this technique were the presence of a large bone fragment, and palmar subluxation or the loss of joint congruity of the distal interphalangeal joint. Using the Wehbe and Scheider classification there were 27 type IB, 19 type IIB, 17 type IA, and 2 type IIA fractures. According to the Crawford rating system there were 46% excellent, 32% good, 20% fair and 2% poor results. We believe that this technique, when properly applied, produces satisfactory results. PMID- 12531663 TI - Anterolateral thigh free flap for complex soft tissue hand reconstructions. AB - The anterolateral thigh free flap is described in terms of its surgical anatomy, techniques of elevation and its application in seven cases of trauma to the hand. The particular advantages of this free flap for reconstructing a variety of hand defects are demonstrated. Flap refinements that have potential applications for hand surgery are described. PMID- 12531664 TI - Restoration of pinch grip in ulnar nerve paralysis: extensor carpi radialis longus to adductor pollicis and abductor pollicis longus to first dorsal interosseus tendon transfers. AB - A double tendon transfer was used to restore thumb pinch in nine consecutive patients. The extensor carpi radialis longus was transferred to the adductor pollicis tendon and one slip of the abductor pollicis longus was transferred to the first dorsal interosseus tendon. The patients were followed for 6 (range, 2 10) years and there were no instances of transfer rupture. Selective recruitment of the motors of the transfers was possible in all cases. Their amplitude was considered adequate for all the cases of adductor transfers but was limited in all of the first dorsal interosseus ones. Key pinch was 73% (range, 41-104%), the pulp-to-pulp pinch was 72% (range, 50-95%) and the power grip was 73% (range, 35 91%) of the opposite hand. The force of thumb adduction was 63% (range, 27-132%) and of index finger abduction was 58% (range, 21-104%) of the unaffected side. PMID- 12531665 TI - Dupuytren'S disease affecting the thumb and first web of the hand. AB - The clinical distribution of the Dupuytren's disease cords in the thumb and first web was examined in 100 consecutive patients with Dupuytren's disease. The precise anatomical relations of the cords were then studied in 25 patients undergoing Dupuytren's surgery for thumb and first web space disease. Thumb and first web space involvement was found in 28% of hands affected by Dupuytren's disease, and was the third most common site after the ring and little fingers. Operative findings showed that two major cords could be distinguished. One lying in the first web space and passing towards the insertion of the first dorsal interosseous muscle on the radial side of the index finger and the other lying on the radial aspect of the thumb. PMID- 12531666 TI - Intramuscular tenotomy of flexor digitorum superficialis in the distal forearm after surgical excision of dupuytren's disease. AB - Contracture of the proximal interphalangeal joint after surgery to excise Dupuytren's disease, despite release of the contributory structures within the finger, can be caused by flexor digitorum superficialis (FDS) contracture. We describe five cases where FDS contracture was released by intramuscular tenotomy in the distal forearm. Standard postoperative therapy for Dupuytren's fasciectomy was used and clinical review showed improved finger extension with no loss of strength. We suggest that intramuscular tenotomy of FDS in the forearm can be used safely where indicated after excision of the Dupuytren's disease. PMID- 12531667 TI - Obstetric brachial plexus lesions: a study of 74 children registered with the British Paediatric Surveillance Unit (March 1998-March 1999). AB - Seventy-four children with obstetric brachial plexus palsy registered with the British Paediatric Surveillance Unit were prospectively followed for a minimum of 2 years. Thirty-nine (52.7%) spontaneously recovered to normal or nearly normal levels and a further 29 (39.3%) regained good function in the upper limb. The most important secondary deformity involved the gleno-humeral joint and 20 patients (27%) needed surgical correction. Two more children await operation for shoulder deformity. The brachial plexus was explored in nine patients (12.2%) and repaired in seven. PMID- 12531668 TI - Assessment of the motor power in older children with obstetric brachial plexus palsy. AB - The King Saud University (KSU) muscle grading system used for assessing the upper limb in older children with obstetric brachial plexus palsy is presented and compared to other muscle grading systems. PMID- 12531669 TI - Minimally displaced Colles' fractures: a prospective randomized trial of treatment with a wrist splint or a plaster cast. AB - Sixty-six adult patients with minimally displaced distal radial fractures were randomly assigned to treatment with either a plaster cast or a lightweight removable wrist splint. Outcome assessment was by clinical and radiological evaluation and an independent physiotherapy assessment. There were significant differences between the treatments for cast satisfaction, cast problems and the functional assessment score at 6 weeks, with the removable splint scoring better in all cases. PMID- 12531670 TI - Preliminary results of total wrist arthroplasty using the RWS Prosthesis. AB - Twenty-nine wrist prostheses were implanted and followed-up for a mean of 4 (range, 2-8) years. Twenty-four of these patients had rheumatoid arthritis. Pain and range of motion improved significantly. Sixteen patients were graded as excellent, six as good, two as fair and three as poor according to Meuli's wrist score. There were three frank radiological failures and one revision in this study. Eleven prostheses showed signs of possible loosening without progression or clinical instability. The management of the distal ulna during this procedure and the long-term fixation of the carpal component were problematic. PMID- 12531671 TI - Fate of failed Meuli total wrist arthroplasty. AB - Sixteen Meuli wrist arthroplasties in 13 patients suffering mainly from rheumatoid arthritis were revised for failure. Causes for failure were mechanical problems with the implant in three wrists, soft tissue problems in two wrists and a combination of mechanical failure and soft tissue problems in 11 wrists. Management included 11 revision arthroplasties in ten wrists, four arthrodeses and two primary soft tissue reconstructions. Twenty-six additional soft tissue procedures were required in association with the revision arthroplasties or arthrodeses. Five of the 11 revision arthroplasties had to be converted to arthrodeses after an average of 5 (range 3-8) years. After removal of a failed wrist implant union of the salvage arthrodesis was difficult to achieve in two of the nine instances. This series demonstrates that revision arthroplasty may be a useful alternative to arthrodesis for the salvage of primary wrist arthroplasties in rheumatoid patients. However, complications and reoperations may occur after both revision arthroplasty and arthrodesis. PMID- 12531672 TI - Wrist function after excision of the pisiform. AB - The pisiform bone may provide mechanical stability to the ulnar column of the wrist by preventing triquetral subluxation. Thus, surgical excision of the pisiform might cause loss of function to the wrist. We performed a functional evaluation of 20 hands in 20 patients who had undergone pisiformectomy for pisotriquetral joint dysfunction. At a mean follow-up of 65 months, 15 patients had complete relief of symptoms and five continued with mild discomfort. Compared with the unaffected wrist, there were no significant differences in grip strength and wrist movement, static strength and dynamic power. We conclude that pisiformectomy for pisotriquetral joint dysfunction can be safely recommended because restoration of function with a painless wrist is the normal outcome. PMID- 12531673 TI - Anterior subfascial transposition of the ulnar nerve. AB - After anterior subfascial transposition, the ulnar nerve lies superficial to the flexor-pronator muscle group but deep to its fascia. Eight patients with cubital tunnel syndrome were treated with this method and reviewed retrospectively. The average age at the time of operation was 52 years. All patients had severe cubital tunnel syndrome based on Dellon's classification. The average follow-up period was 2 years and 9 months. Post-operative outcome assessment was based on the modified Bishop rating system. Six patients had excellent and two had good outcomes. All were back at work by the 5th post-operative week. There were no complications or recurrence of symptoms. Anterior subfascial transposition of the ulnar nerve is an effective method of surgical treatment for patients with severe cubital tunnel syndrome. PMID- 12531674 TI - Carpal tunnel syndrome - is high-frequency ultrasound a useful diagnostic tool? AB - This study assessed the clinical use of three ultrasound measurements; median nerve cross-sectional area, median nerve flattening ratio and palmar displacement of the flexor retinaculum, for the diagnosis of carpal tunnel syndrome. The measurements were made in 20 carpal tunnel sufferers and 20 controls. The sensitivity, specificity and predictive values of each were calculated in various clinical settings. Values for each of the three variables were significantly different in the patient and control populations. The differences we recorded were smaller than those found in previous studies. The tests had a sensitivity of 72% and a specificity of 90%. Alterations in the morphology of the carpal tunnel in patients with carpal tunnel syndrome can be measured in the district general hospital setting. The measurements described maybe a useful non-invasive confirmatory test in patients in whom there is a strong clinical suspicion of carpal tunnel syndrome. However, they would be of no benefit in epidemiological surveys of populations with a low incidence of carpal tunnel syndrome. PMID- 12531675 TI - Hemiarthroplasty for trapeziometacarpal arthritis - a useful alternative? AB - We performed hemiarthroplasty using the Swanson titanium implant for treatment of isolated and advanced trapeziometacarpal joint osteoarthritis Nineteen implants were placed in 18 patients (mean age: 57 years) between 1995 and 1999. One joint required revision and conversion to trapeziectomy. Eighteen joints in 17 patients were evaluated with a mean follow-up period of 34 (range, 14-60) months. Good pain relief was noted in 13 hands. All patients had good hand function as scored using validated questionnaires. Radiographic loosening occurred with five implants and trapezial wear in ten joints. Though radiographic loosening or trapezial wear did not correlate with less satisfactory clinical results, failure of the implant is a concern in the long term. Preliminary results indicate that hemiarthroplasty can be a useful treatment alternative in selected, relatively young patients with isolated trapeziometacarpal osteoarthritis and good bone stock. Good motion and stability can be preserved Failures can be effectively salvaged by trapeziectomy. PMID- 12531676 TI - A comparison of ex vivo and in vitro Sutter metacarpophalangeal prostheses. AB - Forty-one Sutter metacarpophalangeal prostheses were implanted into 11 hands of nine patients. Twelve of these prostheses were revised from three patients after a mean period of 42 months. Of the 12 prostheses, 11 showed fracture at the junction of the distal stem and the central hinge region. Two Sutter metacarpophalangeal prostheses were tested on a single station finger simulator and both failed due to fracture at the junction of the hinge and the distal stem. PMID- 12531677 TI - The two-level ulnar collateral ligament injury of the metacarpophalangeal joint of the thumb. AB - Ulnar collateral ligament injuries of the thumb may present as avulsion fractures of the bone or as ligamentous tears. Displaced fracture fragments or Stener lesions demand operative management. Occasionally, both fracture and ligamentous tears may occur simultaneously, and the management of the ligamentous aspect may be overlooked in cases with undisplaced fractures leading to failure of non operative treatment. It is important in cases with a relatively undisplaced fracture fragment to exclude a simultaneous ligamentous tear and Stener lesion by careful clinical examination and by examining the radiographs for telltale displaced flecks of bone. PMID- 12531678 TI - Osteochondral grafting of the metacarpophalangeal joint in rheumatoid arthritis. AB - We report two cases of isolated metacarpophalangeal joint subluxation in rheumatoid patients with dorsal osteochondral defects on the base of the proximal phalanx. These were treated by open reduction, autologous osteochondral grafting and synovectomy. Stable reduction was maintained and appeared to delay the deterioration of the joint. PMID- 12531681 TI - Apoptosis in lymph nodes and changes in lymphocyte subpopulations in peripheral blood of pigs infected with porcine rubulavirus. AB - In a first experiment, five pigs were inoculated intranasally with porcine rubulavirus (PoRV) at 5 days of age and killed 7 days post-infection (pi). In a second experiment, four pigs were infected with the same virus at 17 days of age and killed at 9 or 15 days pi. Control piglets in each experiment received uninfected cell culture supernate. All PoRV-infected pigs developed respiratory and nervous signs, and histological lesions of non-suppurative encephalitis and interstitial pneumonia. All control pigs remained clinically normal and did not have histological lesions. Significantly increased numbers of apoptotic cells were detected by terminal deoxynucleotidyl transferase biotin-dUTP nick end labelling (TUNEL) in tonsil and lymph nodes of the pigs infected at 7 days of age and killed at 7 days pi. Significantly increased percentages of CD2(+) and CD8(+) T lymphocytes were also found in peripheral blood of these animals at this time, while the percentages of CD4(+) and MHC class II lymphocytes were significantly reduced. Significantly increased numbers of apoptotic cells were detected in lymphoid tissues of the pigs infected at 17 days of age and killed at 9 days pi. The percentages of CD2(+), CD8(+) and MHC class II lymphocytes in peripheral blood were also significantly increased at this time; the percentage of MHC class II lymphocytes remained elevated at 15 days pi. These results indicate that induction of apoptosis is an important mechanism in the pathogenesis of PoRV infection in young pigs, and that this virus induces changes in lymphocyte subpopulations in peripheral blood. PMID- 12531682 TI - Abnormal neuronal expression of the calcium-binding proteins, parvalbumin and calbindin D-28k, in aged dogs. AB - Disturbances of the gamma-aminobutyric acid (GABA) neurotransmitter system have been implicated in chronic degenerative neurological disease. Cognitive dysfunction and neuron loss are features in older dogs. GABAergic neurons also show immunoreactivity for specific calcium-binding proteins. Immunohistochemistry was used to study the neuronal expression of calbindin D-28k and parvalbumin in different areas of the brain in 13 dogs, aged between 2 and 13.5 years. Calbindin expression was found only in the cerebellum. There were significant differences in the quantity and distribution of neurons expressing these proteins between geriatric and adult brains. Parvalbumin- and calbindin-expressing neurons are relatively sensitive to degeneration in the cerebellum of older dogs. Parvalbumin labelling was associated with dystrophic structures that are commonly associated with ageing. PMID- 12531683 TI - Separation of bovine plasma lipoproteins by a rapid ultracentrifugation method. AB - The recently described method of centrifugation with iodixanol for the rapid separation of human plasma lipoproteins was adapted to separate bovine plasma lipoproteins. Density gradients were generated by mixing plasma with iodixanol 12% (w/v), followed by centrifugation at 350,000 g and 16 degrees C for 3 h 10 min in a vertical rotor. Gradients were unloaded dense-end first into 10 fractions. Human very low density lipoprotein (VLDL; density < 1.011 g/ml), low density lipoprotein (LDL; density = 1.016-1.039 g/ml) and high density lipoprotein (HDL; density = 1.039-1.090 g/ml) were resolved well at densities considerably lower than those traditionally reported in salt gradients. In gradients generated from 12% iodixanol, bovine LDL and HDL exhibited even lower densities (1.016-1.028 and 1.016-1.048 g/ml, respectively) with all lipoproteins occurring at the lower density region of the gradient. In contrast, density gradients generated from layers of equal volumes of 6% and 12% iodixanol readily separated bovine HDL from VLDL, whilst LDL still overlapped with HDL. The latter accounts for >80% of all bovine lipoproteins and exists as two populations, namely light and heavy HDL. Gradients generated from two layers of iodixanol recovered bovine HDL in five fractions. The hypercholesterolaemia associated with lactation resulted in a modest shift in the profile of HDL cholesterol towards lipoprotein particles of lower density (light HDL). Significant between-farm differences were also detected in the density profiles of bovine plasma cholesterol. This new method is suitable for use in research and diagnosis in relation to lipoprotein metabolism disorders in cows. PMID- 12531684 TI - Immune complex-associated thrombocytopenic purpura syndrome in sexually mature Gottingen minipigs. AB - Eleven cases of thrombocytopenic purpura (TP) in sexually mature male or female Gottingen minipigs occurred sporadically over 3 1/2 years in a closed breeding colony protected by strict barrier conditions. Typical clinical signs of TP, including extensive subcutaneous haemorrhages, were seen in all affected animals. Haematological abnormalities included marked thrombocytopenia and anaemia. A consistent histopathological finding was the presence of membranoproliferative lesions in the renal glomeruli. Immunohistochemically, glomerular deposits were positively labelled for complement factor C1q and often also for immunoglobulins. Bone marrow findings consisting of increased numbers of immature and apoptotic megakaryocytes were compatible with a state of increased platelet consumption. Based on the combined presence of thrombocytopenia and renal immune complexes, it is suggested that the syndrome was related to a type III hypersensitivity reaction. However, further studies are needed to verify this hypothesis. PMID- 12531685 TI - Chemotactic response of lymphocytes in chicken embryos infected with Mycoplasma gallisepticum. AB - A prominent feature of Mycoplasma gallisepticum (MG) infection is a lymphoproliferative response at the site of infection. In this study, artificial air cells (AACs) were made in eggs containing 16-day old chicken embryos. An MG culture and supernates from MG-infected RP-9 cells, HD-11 cells and monocytes were separately deposited on the membranes of the AAC. After incubation for 5 days, the eggs were opened and the AAC membranes were collected for histopathological examinations. Immunolabelling of MG-infected membranes showed a massive infiltration of lymphocytes possessing CD3 surface markers and the presence of cells that secreted lymphotactin, a chemokine. The supernates from MG infected cells caused the infiltration of comparatively small numbers of lymphocytes. The culture medium-inoculated AACs had no obvious abnormalities of the membranes. It is suggested, therefore, that MG infection of the embryonic membranes causes the embryos to secrete lymphotactin, which induces the migration and accumulation of lymphocytes to the sites of infection. PMID- 12531686 TI - Emergence of different macrophage populations in hepatic fibrosis following thioacetamide-induced acute hepatocyte injury in rats. AB - Macrophages may play a role in fibrogenesis. The kinetics and distribution of different macrophage populations were investigated immunohistochemically in hepatic lesions following acute hepatocyte injury induced in F344 rats by a single injection of thioacetamide (TAA) (300 mg/kg body weight, intraperitoneally). Hepatocyte degeneration or necrosis induced by TAA occurred mainly in the perivenular areas of hepatic lobules as early as post-injection (PI) days 1 and 3; fibrotic lesion development began in the damaged areas on day 1, and peaked on day 5; thereafter (PI days 7 and 10), the fibrotic areas decreased and were replaced by regenerated hepatocytes on PI days 15 and 20, indicating a remodelling process. In this rat model, the number of macrophages reacting with ED1 antibody (specific for exudate macrophages), ED2 (recognizing cell membrane antigens of resident macrophages, including Kupffer cells) and OX6 (recognizing MHC class II antigens expressed in antigen-presenting macrophages and dendritic cells) began to increase on PI day 1, peaking on PI day 3. The numbers gradually decreased on PI days 5 and 7; however, the statistically significant increase was maintained in respect of ED1-positive cells up to PI day 20, whereas no significant increase in ED2- and OX6-positive cells remained from PI day 10 onwards. Interestingly, of the ED1-, ED2- and OX6-positive cells, the OX6-positive cells were the least numerous. ED1- and OX6-positive cells appeared exclusively in the injured perivenular areas, whereas ED2-positive cells were present mainly in the mid-zonal areas and in smaller numbers in the perivenular areas. These findings indicated differences in kinetics and distribution between macrophage populations appearing in hepatic fibrosis. In addition, RT-PCR revealed that mRNA expression of osteopontin, a factor for induction and maintenance of macrophages in inflammation, was markedly increased on PI days 5, 7 and 10, suggesting a role in the pathogenesis of hepatic fibrosis. PMID- 12531687 TI - Pathogenesis of postweaning multisystemic wasting syndrome reproduced by co infection with Korean isolates of porcine circovirus 2 and porcine parvovirus. AB - Thirty-two colostrum-deprived conventional pigs aged 28 days were co-infected with Korean isolates of porcine circovirus 2 (PCV2) and porcine parvovirus (PPV) and observed for up to 35 days post-inoculation (dpi). Histopathologically, granulomatous inflammation, with or without intracytoplasmic inclusion bodies, was present in lymphoid tissues (e.g., lymph nodes, thymus, spleen and tonsil) from 20 dpi, being most severe at 24 dpi. PCV2 and PPV DNA were detected in the lymph nodes from 3 to 35 dpi by in-situ hybridization, but the labelling for both viruses was particularly intense and widespread at 20 and 24 dpi. A close relationship between the cells labelled for PCV2 and those labelled for PPV was revealed by examination of serial sections from lymph nodes, and PCV2 and PPV were also detected in peripheral blood monocytes from 3 to 35 dpi. Other tissues and cells in which PCV2 and PPV DNA were detected included macrophages in the tonsil, thymus, spleen, lung, liver, kidney and heart. Significantly more PCV2 positive cells than PPV-positive cells were detected in the lymph nodes at 5 to 35 dpi. The pathogenesis of postweaning multisystemic wasting syndrome (PMWS) reproduced in this study may be suggested thus: initial viral entry through tonsillar macrophages, followed within 3 days by viraemia; PCV2 and PPV replicate, at least to some extent, in circulating peripheral monocytes, contributing to the cell-associated viraemia and to viral distribution throughout the lymphoid tissues. PMID- 12531689 TI - Inner ear morphology in a bilaterally deaf Dogo Argentino pup. AB - Two bilaterally deaf and three unilaterally deaf pups were identified from a litter of 10 Dogo Argentino pups presented for hearing evaluation by electrophysiological investigation. One pup, a bilaterally deaf female aged 43 days, was available for histopathology. Examination of both inner ears revealed bilateral cochlear degeneration with atrophy of the stria vascularis, collapse of the cochlear duct, degeneration of the organ of Corti, and abnormal tectorial membrane. The left vestibule, including the sacculus, was normal. The spiral and vestibular ganglia were essentially normal. This is the first histopathological description of lesions associated with deafness in a Dogo Argentino, but abnormalities were similar to those previously described in deaf Dalmatian pups and in other white hair-coated breeds. The defect was classified as a cochleosaccular degeneration. It was probably congenital and genetic causes were suspected. PMID- 12531688 TI - Detection of classical swine fever virus in the ovaries of experimentally infected sows. AB - Six sows were infected intranasally with a Korean isolate of classical swine fever virus (CSFV). The distribution of virus in ovarian tissues was then assessed for 21 days by in-situ hybridization and immunohistochemistry. CSFV was detected in the ovaries between 7 and 21 days post-inoculation (dpi) by both methods, but the labelling was particularly intense and widespread at 7 dpi. CSFV nucleic acid and antigen were located almost exclusively within the cytoplasm of cells shown by haematoxylin and eosin staining to be macrophages, which were numerous in atretic follicles. Small numbers of CSFV nucleic acid-positive cells with distinctly round morphology and oval nuclei, resembling monocytes, were also observed in the blood vessels of sows at 7 and 14 dpi. CSFV nucleic acid and antigen were not observed in primordial, primary or secondary follicles from infected sows at 7, 14 or 21 dpi. The results suggest that CSFV replicates in circulating peripheral monocytes and gains access to ovarian tissues from the bloodstream, and that this contributes to the distribution of CSFV in macrophages throughout the atretic follicles. PMID- 12531690 TI - Primary malignant mixed mesenchymal tumour of the heart in a dog. AB - A case of primary malignant mixed mesenchymal tumour of the heart in an 8-year old golden retriever is described. The cardiac tumour, measuring 2.5 x 3 x 5 cm, was located in the posterior part of the atrial septum, extending into the surrounding region. Histologically, the tumour was composed of multiple mesenchymal elements of fibrosarcoma, rhabdomyosarcoma, liposarcoma and chondrosarcoma. No previous reports of such a tumour occurring in the heart of the dog were found in the literature. PMID- 12531691 TI - Malignant retroperitoneal paraganglial tumour in a cow. AB - A large primary retroperitoneal sublumbar neoplasm was identified in an 11-year old Holstein cow, with metastases to the lungs, kidneys and lymph nodes. The tumour cells proliferated in a characteristic endocrine pattern, were argyrophilic and positive for neuron-specific enolase, and had membrane-bounded intracytoplasmic granules. In addition, the cells were occasionally positive for cytokeratin and had desmosome-like intercellular junctions. The primary tumour mass was diagnosed as a malignant paraganglial tumour of the aortico-sympathetic ganglion (organ of Zuckerkandl), and was considered to contain primitive cells with epithelial differentiation. PMID- 12531692 TI - Regulation of tumor suppressors by nuclear-cytoplasmic shuttling. AB - Tumor suppressor proteins control the proliferation and survival of normal cells; consequently, their inactivation by gene mutations can initiate or drive cancer progression. Most tumor suppressors have been identified by genetic screening, and in many cases their function and regulation are poorly understood. Ten such proteins were recently shown to contain nuclear transport signals that facilitate their "shuttling" between the nucleus and cytoplasm. This type of dynamic intracellular movement not only regulates protein localization, but also often impacts on function. Here, we review the pathways by which tumor suppressors such as APC, p53, VHL, and BRCA1 cross the nuclear envelope and the impact of regulated nuclear import/export on protein function. PMID- 12531693 TI - NADPH-oxidase activation in murine neutrophils via formyl peptide receptors. AB - Neutrophils play a key role at inflammatory sites where, in addition to destroying infecting microorganisms, they may also have deleterious effects on host tissues. Both activities involve activation of the NADPH-oxidase that produces bactericidal and tissue-destructive reactive oxygen species (ROS). We activated the murine NADPH-oxidase using different types of neutrophil activators and characterized the oxidative responses with respect to magnitude, localization, and kinetics. We show that agonist-induced activation of murine neutrophils results exclusively in extracellular release of ROS and no intracellular production could be detected. We also show that the formylated peptide, formyl-Met-Leu-Phe (fMLF), is a much less potent activator of the murine NADPH-oxidase than of the human analogue. Nevertheless, fMLF responses can be primed by pretreating the murine neutrophils with either cytochalasin B or bacterial lipopolysaccharide. Finally, we show that a synthetic hexapeptide, WKYMVM, is a more potent stimulus than fMLF for murine neutrophils and that these two agonists probably act via nonidentical high-affinity receptors. PMID- 12531695 TI - TGF-beta1-mediated fibroblast-myofibroblast terminal differentiation-the role of Smad proteins. AB - It is now clear that resident myofibroblasts play a central role in the mediation of tissue fibrosis. The aim of the work outlined in this study is to increase our understanding of the mechanisms which drive the phenotypic and functional changes associated with the differentiation process. We have used an in vitro model of transforming growth factor-beta1 (TGF-beta1)-induced pulmonary fibroblast myofibroblast differentiation to examine the role of the TGF-beta1 Smad protein signaling intermediates, in alterations of fibroblast phenotype and function associated with terminal differentiation. TGF-beta1 induced marked alteration in cell phenotype, such that cells resembled "epithelioid-postmitotic fibroblasts." This was associated with marked reorganization of the actin cytoskeleton and upregulation of alphaSMA gene expression. TGF-beta1 stimulation also induced alphaSMA protein expression with increased incorporation of alphaSMA into stress fibers. Following stimulation with TGF-beta1, subsequent addition of serum-free medium did not reverse TGF-beta1-induced morphological change, suggesting that TGF-beta1 induced a relatively stable alteration in fibroblast cell phenotype. Functionally, these phenotypic changes were associated with induction of type I, type III, and type IV collagen gene expression and an increase in the concentrations of the respective collagens in the cell culture supernatant. The role of Smad proteins in terminal differentiation of fibroblasts was examined by transfection of cells, with expression vectors for the TGFbeta1 receptor regulated Smads (R-Smads) or the co-Smad, Smad 4. Transfection with Smad2 but not Smad3 resulted in TGF-beta1 independent alteration in fibroblast cell phenotype, up-regulation of alphaSMA mRNA and reorganization of the actin cytoskeleton. Transfection with Smad4 also induced alteration in cell phenotype, although this was not as pronounced as the effect of overexpression of Smad2. Overexpression of the Smad2, Smad3, or Smad4 proteins was associated with increased production of all collagen types. The study suggests that the phenotypic and functional changes associated with TGF-beta1-induced fibroblast terminal differentiation are differentially regulated by Smad proteins. PMID- 12531694 TI - Downregulation of p21(waf/cip-1) mediates apoptosis of human hepatocellular carcinoma cells in response to interferon-gamma. AB - There is no effective treatment for advanced hepatocellular carcinoma (HCC). We therefore explored the molecular mechanisms of interferon-gamma (IFN-gamma) mediated growth regulation in human HCC cell lines. IFN-gamma receptor expression, signal transduction, and regulation of effectors were examined by RT PCR, immunoprecipitation, immunoblotting, and reporter gene assays. Growth and apoptosis were determined based on cell numbers, cell cycle analyses, kinase assays, DNA fragmentation, and PARP cleavage. HCC cell lines express functionally intact IFN-gamma receptors and downstream effectors. IFN-gamma profoundly inhibited growth of HCC cells via two different mechanisms: inhibition of G1 cell cycle progression and induction of apoptosis. Analyses in SK-Hep-1 cells revealed a deficient cyclin D induction in IFN-gamma-treated cells, resulting in reduced activity of CDK4 and CDK2 kinases and pRB hypophosphorylation. In contrast, apoptosis prevailed in IFN-gamma-treated HepG2 cultures. A survey of apoptosis relevant IFN-gamma effectors including IRF-1, caspase-1, caspase-3, and p21(waf/cip-1) documented a dramatic transcriptional downregulation of p21(waf/cip-1) exclusively in apoptosis-susceptible HepG2 cells. Reconstitution of p21(waf/cip-1) rescued HepG2 cells from IFN-gamma-induced apoptosis, indicating that p21(waf/cip-1) reduction was required for apoptosis execution. Inversely, downregulation of p21(waf/cip-1) sensitized SK-Hep-1 cells to IFN gamma-induced apoptosis. Thus, downregulation of p21(waf/cip-1) in HCC cells functions as a novel, critical determinant of alternative growth inhibitory pathways in response to IFN-gamma. PMID- 12531696 TI - Notch2 protein distribution in human teeth under normal and pathological conditions. AB - Notch signaling is essential for the appropriate differentiation of many cell types during development and, furthermore, is implicated in a variety of human diseases. Previous studies have shown that although the Notch1, -2, and -3 receptors are expressed in developing and injured rodent teeth, Notch2 expression was predominant after a lesion. To pursue the role of the Notch pathway in tooth development and disease, we have analyzed the expression of the Notch2 protein in embryonic and adult wounded human teeth. During the earlier stages of tooth development, the Notch2 protein was expressed in the epithelium, but was absent from proliferating cells of the inner enamel epithelium. At more advanced stages, Notch2 was expressed in the enamel-producing ameloblasts, while it was absent in mesenchyme-derived odontoblasts that synthesize the dentin matrix. Although Notch2 was not expressed in the pulp of adult intact teeth, it was reexpressed during dentin repair processes in odontoblasts and subodontoblastic cells. Transforming growth factor beta-1, which stimulates odontoblast differentiation and hard tissue formation after dental injury, downregulated Notch2 expression in cultured human dental slices, in vitro. These observations are consistent with the notion that Notch signaling is an important element in dental physiological and pathogenic conditions. PMID- 12531697 TI - Regulation of human erythropoiesis by activin A, BMP2, and BMP4, members of the TGFbeta family. AB - Activin A, BMP2, and BMP4, members of the TGFbeta family, have been implicated in the regulation of hematopoiesis. Here we explore and compare, for the first time in human primary cells, the role of activin A, BMP2, and BMP4 during erythropoiesis. Using in vitro erythroid differentiation of CD34(+) primary cells, we obtained the main stages of early erythropoiesis, characterized at the molecular, biochemical, and functional levels. Our results indicate that BMP2 acts on early erythroid cells and activin A on a more differentiated population. We report an insight into the mechanism of commitment of erythropoiesis by activin A and BMP2 involving two key events, increase in EPO-R and decrease in GATA2 expression. Simultaneous addition of activin A with BMP molecules suggests that BMP2 and BMP4 differently affect activin A induction of erythropoiesis. Follistatin and FLRG proteins downmodulate the effects of activin A and BMP2 on erythroid maturation. PMID- 12531698 TI - Ligand- and kinase activity-independent cell survival mediated by the epidermal growth factor receptor expressed in 32D cells. AB - To investigate the intrinsic activities of the epidermal growth factor receptor and the role of its kinase domain in these functions within a cellular environment lacking endogenous ErbB protein expression, wild-type EGF receptor (WT-EGFR) and two kinase-impaired mutants, D813A and K721R, were expressed in 32D murine hematopoietic cells, a line which is normally dependent on interleukin 3 (IL3) for growth and survival. Addition of EGF in the absence of IL3 stimulates receptor autophosphorylation and, in the presence of serum, mitosis in cells expressing WT-EGFR, but not in cells expressing D813A or K721R. Unexpectedly, cells expressing WT-EGFR or K721R exhibited IL3-independent survival in the presence of fetal bovine serum; parental 32D cells and cells expressing D813A did not survive, apparently undergoing apoptosis in the absence of IL3, whether or not serum was present. Addition of EGF did not prevent the apoptosis of WT-EGFR or K721R cells in serum-free medium. Activation of Akt was not necessary to mediate the prosurvival activity of EGF receptor expression. These results suggest that the EGF receptor can mediate the prevention of apoptosis independently of both receptor-ligand binding and receptor kinase activity, and this activity is disrupted by the D813A mutation. PMID- 12531699 TI - Regulation of fatty acid synthase expression in breast cancer by sterol regulatory element binding protein-1c. AB - Activation of fatty acid synthase (FAS) expression and fatty acid synthesis is a common event in human breast cancer. Sterol regulatory element binding proteins (SREBPs) are a family of transcription factors that regulate genes involved in lipid metabolism, including FAS. SREBP-1c expression is induced in liver and adipose tissue by insulin and by fasting/refeeding and is critical for nutritional regulation of lipogenic gene expression. In contrast, upregulation of fatty acid metabolism during in vitro transformation of human mammary epithelial cells and in breast cancer cells was driven by increased MAP kinase and PI 3 kinase signaling, which increased SREBP-1 levels. SREBP-1a was more abundant than SREBP-1c in many proliferative tissues and cultured cells and was thus a candidate to regulate lipogenesis for support of membrane synthesis during cell growth. We now show that SREBP-1c and FAS mRNA were both increased by H-ras transformation of MCF-10a breast epithelial cells and were both reduced by exposure of MCF-7 breast cancer cells to the MAP kinase inhibitor, PD98059, or the PI 3-kinase inhibitor, wortmannin, while SREBP-1a and SREBP-2 showed less variation. Similarly, the mRNA levels for FAS and SREBP-1c in a panel of primary human breast cancer samples showed much greater increases than did those for SREBP-1a and SREBP-2 and were significantly correlated with each other, suggesting coordinate regulation of SREBP-1c and FAS in clinical breast cancer. We conclude that regulation of FAS expression in breast cancer is achieved through modulation of SREBP-1c, similar to the regulation in liver and adipose tissue, although the upstream regulation of liopgenesis differs in these tissues. PMID- 12531701 TI - Phytoremediation of selenium by two helophyte species in subsurface flow constructed wetland. AB - The phytoremediation of selenium by two different wetland species was investigated. Selenium (20.4 microg/l) was supplied continuously to subsurface flow constructed wetlands, one vegetated with Typha latifolia L. and the other with Phragmites australis (Cav.) Trin. ex Steud. The beds of both species had same hydraulic loading rate (0.079 m(3)/m(2)/d) and water retention time (24 h). However, the mass loading rate was 1.27 mg Se/m(2)/d for Phragmites and 1.35 mg Se/m(2)/d for Typha. In the Typha bed Se migrated faster than in the Phragmites bed. After 25 d of Se supplementation in the Typha bed about 54% of the Se inlet concentration remained in the outlet water. In the Phragmites bed Se was removed completely from the water after passing through 3/4 of the bed length. After 65 d of Se supplementation the highest amount of Se (2.8 microg/g dry matter) was determined in the organic material of the Typha bed. Roots and rhizomes accumulated 2.2 and 1.8 microg/g dry matter respectively. Phragmites accumulated Se in the leaves and stems, but not in the rhizomes. The accumulation in the leaves (1.8 microg Se/g dry matter) was three times higher than in the stems (0.6 microg Se/g dry matter). PMID- 12531700 TI - Reduction of the phenolic components in olive-mill wastewater by an enzymatic treatment and its impact on durum wheat (Triticum durum Desf.) germinability. AB - Olive-mill wastewater (OMW), an effluent of olive oil extraction process, is annually produced in huge amounts in olive growing areas. An interesting option for its disposal is the spreading on agricultural land, provided that phytotoxic effects are neutralized. The objective of the present investigation was to evaluate the potential of an enzyme-based treatment in removing OMW phytotoxicity. To this aim, germinability experiments on durum wheat (Triticum durum Desf. cv. Duilio) were conducted in the presence of different dilutions of raw or enzyme-treated OMW. OMW treatment with laccase resulted in a 65% and 86% reduction in total phenols and ortho-diphenols respectively, due their polymerization as revealed by size-exclusion chromatography. Raw OMW exerted a significant concentration-dependent inhibition on the germinability of durum wheat seeds which was evident up to a dilution rate of 1:8. When the effluent was treated with a fungal laccase, germinability was increased by 57% at a 1:8 dilution and by 94% at a 1:2 dilution, as compared to the same dilutions using untreated OMW. The treatment with laccase also decreased the mean germination time by about 1 day as compared to untreated controls. These results show that germinability inhibition due to OMW can be reduced effectively using fungal laccase, suggesting that phenols are the main determinants of its phytotoxicity. PMID- 12531702 TI - Decolorization of structurally different synthetic dyes using cobalt(II)/ascorbic acid/hydrogen peroxide system. AB - The cobalt(II)/ascorbic acid/hydrogen peroxide system was used for decolorization of azo, acridine, anthraquinone, thiazine and triphenylmethane dyes. More than 90% decolorization was obtained with all dyes except Remazol Brilliant Blue R (75%). With other transition metals the system was less efficient. With copper, higher concentration and prolonged incubation time was necessary to obtain the same extent of decolorization. The rate of decolorizaton was not affected by pH in the range of 3-9. The reaction is very fast, with more than 90% decolorization being attained within 15 min. The system produces hydroxyl radicals which are responsible for the decolorization. PMID- 12531703 TI - The direct photolysis and photocatalytic degradation of alachlor at different TiO2 and UV sources. AB - Direct photolysis and photocatalytic degradations of alachlor, a widely used herbicide, were studied using three different monochromatic UV lamps (254, 300 and 350 nm) and two TiO(2) sources. Both the direct photolysis and photocatalytic degradations of alachlor follow pseudo-first-order decay kinetics. TiO(2)-P25 was found to be an effective photocatalyst compared to TiO(2)-BDH. The direct photolysis of alachlor was dominant at 254 nm even if TiO(2) was present in the solution. Among the three UV wavelengths used, the highest photocatalysis quantum yield was obtained at 300 nm. The photocatalytic degradation rate of alachlor increased with the dosages of TiO(2), but an overdose of TiO(2) would retard the reaction due to light attenuation. Photocatalytic reactions were slightly enhanced in an alkaline medium, and the different proton sources causing various degrees of rate retardation were due to the presence of the corresponding counter anions. This effect was diminished at a later stage after the reaction intermediates were formed. PMID- 12531704 TI - Photoelectrocatalytic degradation of 4-chlorophenol and oxalic acid on titanium dioxide electrodes. AB - Photocatalytically active thin TiO(2) films were produced by spin-coating or dip coating an alkoxy precursor onto a transparent conducting electrode substrate and by thermal oxidation of titanium metal. The thin films were used to study the photoelectrocatalytic or photoelectrochemical degradation of oxalic acid and 4 chlorophenol (4-CP) under near UV (monochromatic, 365 nm) light irradiation. Degradation was monitored by a variety of methods. In the course of oxalic acid degradation, CO(2) formation accounted for up to 100% of the total organic carbon degradation for medium starting concentrations; for the degradation of 4-CP, less CO(2) was detected due to the higher number of oxidation steps, i.e. intermediates. Incident-photon-to-current conversion efficiency, educt degradation and product formation as well as Faradaic efficiencies were calculated for the degradation experiments. Quantum yields and Faradaic efficiencies were found to be strongly dependent on concentration, with maximum values (quantum yield) around 1 for the highest concentrations of oxalic acid. PMID- 12531705 TI - The light transmission and distribution in an optical fiber coated with TiO2 particles. AB - The light delivery and distribution phenomena along the optical fiber coated with the P-25 TiO(2) particles by dipping was investigated. The surface properties (coverage, roughness and thickness) of the TiO(2) layer coated on the optical fiber were characterized by SEM micrographs. For TiO(2) layer prepared from solutions containing less than 20 wt.% of TiO(2) slurry, the thickness of layer was increased linearly with the TiO(2) slurry content in solutions. The UV light intensity transmitted along a TiO(2)-coated optical fiber decreased more rapidly than that transmitted along a non-coated fiber. Based on the experimental results, the light intensity distribution around a coated optical fiber was modeled to determine the optimum configuration for the design of optical fiber reactors under various operational conditions. PMID- 12531706 TI - Radiolysis of aqueous phenol solutions with nanoparticles. 1. Phenol degradation and TOC removal in solutions containing TiO2 induced by UV, gamma-ray and electron beams. AB - Aqueous phenol solutions containing TiO(2) nanoparticles were irradiated with ultraviolet (UV), gamma-ray and electron beams. Organic compounds were fully removed by each type of radiation in the presence of the particles. The absorbed energy of the ionizing radiation (gamma-ray and electron beams) needed for removal was much lower than that of UV photocatalysis. Phenol was decomposed by the ionizing radiation in the absence of the nanoparticles and the addition of TiO(2) had no significant effect on phenol decomposition rate. Instead, total organic carbon (TOC) removal using the ionizing radiation was accelerated drastically by TiO(2). It is suggested that TiO(2) particles affect the intermediate compounds produced through the decomposition of phenol. The amount of removed TOC per absorbed energy were compared in the absence and the presence of TiO(2) nanoparticles. Radiolysis with the nanoparticles showed consistently high rate and high efficiency of TOC removal. PMID- 12531707 TI - Sonolysis of alkylphenols in aqueous solution with Fe(II) and Fe(III). AB - The sonolytic degradation of alkylphenols (APs), such as butylphenol, pentylphenol, octylphenol, and nonylphenol (NP), in water was investigated at a sound frequency of 200 kHz with an acoustic intensity of 6 W cm(-2) under argon, oxygen, and air atmospheres. The sonolytic degradation rate of the APs under the conditions of the present study depended upon their alkyl chain length. The decrease in the degradation rate by the radical scavenging effect was in the range of about 48-82% for APs in the presence of 3 mM 2-methyl-2-propanol. The free radicals play a significant role in the sonolytic degradation process of the APs. In the presence of Fe(II) and Fe(III), the pseudo-first-order rate constants for the sonolytic degradation of 30 microM NP as a function of the concentration of Fe(II) and Fe(III) were estimated under argon and oxygen. The maximum rate constants were observed at 50 microM Fe(II) (0.139 +/- 0.008 min(-1)) and 100 microM Fe(III) (0.103 +/- 0.001 min(-1)) under oxygen. The total organic carbon concentration (TOC) was investigated under same conditions. TOC decreased in the range of about 50-70% during the sonication in the presence of Fe(II) and Fe(III) under argon and oxygen. The sonochemical effects by the addition of Fe(II) and Fe(III) as catalyst during the sonication under the proper atmosphere result in a remarkable enhancement of degradation and mineralization. PMID- 12531708 TI - Ozone treatment of soil contaminated with aniline and trifluralin. AB - Column studies were conducted to determine the ability of ozone to degrade aniline and trifluralin in soil. Ozone rapidly degraded aniline from soil under moist soil conditions, 5% (wt). Removal of 77-98% of [UL-14C]-aniline was observed from soil columns (15 ml, i.d. = 2.5 cm), exposed to 0.6% O(3) (wt) at 200 ml/min after 4 min. Initial ozonation products included nitrosobenzene and nitrobenzene, while further oxidation led to CO(2). Ring-labeled-[UL-14C] trifluralin removal rates were slower, requiring 30 min to achieve removals of 70 97%. Oxidation and cleavage of the N-propyl groups of trifluralin was observed, affording 2,6-dinitro-4-(trifluoromethyl)-aniline, 2,6-dinitro-N-propyl-4 (trifluoromethyl)-benzamine, and 2,6-dinitro-N-propyl-N-acetonyl-4 (trifluoromethyl)-benzamine. Base solutions revealed that trifluralin was similarly oxidized to CO(2), where 72-83% of the activity recovered comprised 14CO(2). Use of ozone-rich water improved contaminant removal in trifluralin amended soil columns, but did not improve removal in aniline, pentachloroaniline, hexachlorobenzene amended soil columns, suggesting that ozonated water may improve contaminant removal for reactive contaminants of low solubility. PMID- 12531709 TI - Catalytic reduction of NO on copper/MCM-41 studied by in situ EXAFS and XANES. AB - Speciation of copper in the channels of MCM-41 during reduction of NO with CO at 473-773 K was studied by in situ extended X-ray absorption fine structural (EXAFS) and X-ray absorption near edge structural (XANES) spectroscopies in the present work. The component fitted (in situ) XANES spectra of the catalyst showed that about 72% of metallic copper (Cu(0)) in MCM-41 was oxidized to higher oxidation state coppers (Cu(II) (46%) and Cu(I) (26%)) during the NO reduction process (at 473 K). By EXAFS, we also found that in the NO reduction process, oxygen was inserted into the metallic copper matrix and led to a formation of the copper oxide species with a Cu-O bond distance of 1.93 A which was greater than that of the model compound Cu(2)O (typically 1.86 A). At 573-673 K, mainly Cu(II) was found in the channels of MCM-41. Nevertheless, at a higher temperature (e.g., 773 K), about 61% Cu(I), 31% Cu(II), and 8% Cu(O) with averaged Cu-Cu and Cu-O bond distances of 3.04 and 1.88 A, respectively were observed, that might account for the high selectivity-to-decomposition (S/D) ratios for yields of N(2) and CO(2) in the catalytic reduction of NO with CO. PMID- 12531710 TI - Improvement of DOC removal by multi-stage AOP-biological treatment. AB - The single and multi-stages advanced oxidation process (AOP)-biological treatments were evaluated to apply for drinking water treatment, especially for the water containing less susceptible dissolved organic carbon (DOC) to ozone, comparing with the ozonation-biological treatment. Minaga reservoir water and the secondary effluent from a Municipal wastewater treatment plant were used as dissolved organic matter (DOM) solutions. DOC removals after 60 min AOP biological treatment were 62% and 41% in the Minaga reservoir water and the secondary effluent, respectively, whereas those in the ozonation-biological treatment only 40% and 15% of DOC were removed, respectively. The result indicated that the single-stage AOP-biological treatment could improve DOC removal in comparison with the single-stage ozonation-biological treatment. This is because the AOP mineralized both biodegradable dissolved organic carbon (BDOC) produced in the early stage of oxidation and non-biodegradable dissolved organic carbon (NBDOC), whereas only BDOC was mineralized by further ozonation and NBDOC was not oxidized in the ozonation-biological treatment. The multi-stage treatment could not improve DOC removal in comparison with the single-stage treatment in the ozonation-biological treatment for the secondary effluent containing less susceptible DOC to ozone. However, the multi-stage AOP-biological treatment significantly reduced DOC and achieved 71% of DOC removal by 4 times repetition of 15 min oxidation, whereas DOC removal was 41% in the single-stage AOP biological treatment for the same oxidation time. The improvement of DOC removal by the multi-stage AOP-biological treatment was due to BDOC removal as a radical scavenger by subsequent biological treatment in the early stage of oxidation and direct mineralization in the latter stage of oxidation. PMID- 12531711 TI - Application of Fenton's reagent to regenerate activated carbon saturated with organochloro compounds. AB - In this work Fenton's reagent was used to treat activated carbon saturated with organochloro compounds for the oxidation of the adsorbed contaminants and the regeneration of the carbon adsorbent. Activated carbon containing adsorbed chlorinated model substrates such as chlorobenzene, tetrachloroethylene, chloroform or 1,2-dichloropropane was treated with Fenton's reagent at room temperature resulting in a rapid consumption of the organochloro compounds. Thermogravimetric, infrared, BET surface area and MIMS adsorption studies showed that Fenton's treatment has no significant effect on the physico-chemical properties of the activated carbon. The used carbon adsorbents can be efficiently regenerated and recycled with no loss of its adsorption capacity even after five consecutive treatments with Fenton's reagent. PMID- 12531712 TI - Biotransformation of cyanide to methane and ammonia by Klebsiella oxytoca. AB - Klebsiella oxytoca, isolated from cyanide-containing industrial wastewater, was shown to be able to biodegrade cyanide to non-toxic endproducts using cyanide as the sole nitrogen source. In this study, ammonia was one of the detected endproduct of cyanide biodegradation by the concentrated resting cells of K. oxytoca. Moreover, cyanide has been shown to be biotransformed to methane through the actions of concentrated resting cells. Biodegradation of cyanide by cell-free extracts was not observed, which might be due to the inactivation of nitrogenase (an oxygen-labial enzyme) caused by the oxygen exposure after cell disruption. Results show that the cyanide consumption by resting cells of K. oxytoca was induced when the pretreatment of these cells with cyanide was conducted. However, the cyanide-degrading capability of resting cells pretreated with ammonia was inhibited. The inhibition of cyanide degradation by resting cells of K. oxytoca was affected by the ammonia concentration. This might result from the suppression of nitrogenase activity of K. oxytoca by ammonia since nitrogenase was suggested to be the sole cyanide-degrading enzyme during the cyanide degradation process. Results from this study also show that the processes of cyanide biodegradation and ammonia production by resting cells occurred simultaneously. This suggests that the utilization of cyanide as nitrogen source by K. oxytoca might proceed using ammonia as an assimilatory substrate. PMID- 12531713 TI - Effect of reducing conditions on sludge melting process. AB - The objective of this study was to compare the effects of CO/CO(2) reducing conditions with those of air oxidizing conditions on the pouring temperature of the sludge melting process and the heavy metal leachability of the resultant sludge slag. Synthetic sludge ash composed of SiO(2), CaO and Al(2)O(3), as well as sewage sludge ash generated from a laboratory incinerator was employed. The experimental results indicated that the pouring temperatures are significantly reduced under the reducing conditions of CO/CO(2), or 24 and 77 degrees C lower than under air conditions for synthetic and sludge ash, respectively. The heavy metal leaching tests further indicate lower heavy metal concentrations present in the leachate under the reducing conditions, notably an order of magnitude lower in Zn. However, X-ray diffractogram indicates similar peaks for these two slags produced under different conditions. PMID- 12531714 TI - Biodegradation of 2,4-dichlorophenol in sequencing batch reactors augmented with immobilized mixed culture. AB - 2,4-Dichlorophenol (2,4-DCP) degrading mixed culture was immobilized in polyvinyl alcohol jel beads and supplemented to sequencing batch reactors (SBR) to treat 2,4-DCP containing wastewater. Impacts of bioaugmentation level on the performance of bioaugmented systems were studied. Results show that inoculum size affected the start-up time of the SBR systems. For the non-augmented SBR system, nine days was needed for the system to start-up, whereas it only took six, four, three and two days for the SBRs with 1.9%, 3.7%, 5.6% and 9.3% immobilized culture, respectively. In addition, bioaugmented SBR systems demonstrated stronger capacity to cope with high 2,4-DCP shock loading than the control system. The control SBR failed to treat 2,4-DCP at 166 mg/l in influent, while the SBR with 1.9% inoculation could successfully cope with 2,4-DCP at 166 mg/l, but failed at 250 mg/l, and the SBR with 3.7%, 5.6% and 9.3% immobilized culture could successfully degrade 250 mg/l 2,4-DCP in feed. Furthermore, the contributions to the removal of 2,4-DCP by the introduced and indigenous culture in an augmented SBR system at various operation stages were investigated. It was found that augmented culture played the primary role in degrading 2,4-DCP at the beginning of system start-up, but after one-month operation, both the indigenous and the introduced culture posed strong ability to degrade 2,4-DCP. PMID- 12531715 TI - A comparative study on the biosorption characteristics of some yeasts for Remazol Blue reactive dye. AB - Biosorption capacities and rates of different kinds of dried yeasts (Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces marxianus, Candida sp., C. tropicalis, C. lipolytica, C. utilis, C. quilliermendii and C. membranaefaciens) for Remazol Blue reactive dye from aqueous solutions were compared under laboratory conditions as a function of initial pH and initial dye concentration. Optimum initial biosorption pH was determined as 2 for all the yeasts. All the yeast species showed comparable and very high dye sorption at 100 mg/l initial dye concentration. The equilibrium sorption capacity of the biomass increased with increasing initial dye concentration up to 400 mg/l for Candida sp. C. lipolytica and C. tropicalis; up to 300 mg/l for C. quilliermendii and C. utilis and up to 200 mg/l for S. cerevisiae, S. pombe, K. marxianus and C. membranaefaciens while the adsorption yield of dye showed the opposite trend for all the yeasts. Among the nine yeast species, C. lipolytica exhibited the highest dye uptake capacity (Q(0) = 250 mg/g). Both the Freundlich and Langmuir adsorption models were found suitable for describing the biosorption of the dye by all the Candida yeasts (except C. membranaefaciens). The results indicated that the dye uptake process followed the pseudo-second-order kinetics for each dye-yeast system. PMID- 12531716 TI - Solar activated ozonation of phenol and malic acid. AB - The effect that sunlight has on the degradation rate of two model organic compounds, phenol and malic acid, by ozone is studied. The effect seems to be due to both direct light absorption (300-320 nm photons) by ozone, which produces the pollutant degradation, and light absorption by reaction intermediates. The presence of such a light notably improves the reactivity of ozone toward the organic species, leading to a faster and complete mineralization even at large initial total organic carbon values. The use of artificial sunlight (Xe lamp) is also explored. Finally, the simultaneous presence of sunlight and other ozone degradation catalyst like transition metal ions is studied, showing the beneficial effect of such a combination. PMID- 12531717 TI - Adsorption behavior of reactive dye in aqueous solution on chemical cross-linked chitosan beads. AB - A batch system was applied to study the adsorption of reactive dye (reactive red 189) from aqueous solutions by cross-linked chitosan beads. The ionic cross linking reagent sodium tripolyphosphate was used to obtain more rigid chitosan beads. To stabilize chitosan in acid solutions, chemical cross-linking reagent epichlorohydrin (ECH), glutaraldehyde and ethylene glycol diglycidyl ether was used and ECH shows a higher adsorption capacity. The Langmuir and Freundlich adsorption models were applied to describe the equilibrium isotherms at different particle sizes and isotherm constants were determined. The Langmuir model agrees very well with experimental data and its calculated maximum monolayer adsorption capacity has very large value of 1802-1840 (g/kg) at pH 3.0, 30 degrees C. The kinetics of the adsorption with respect to the initial dye concentration, temperature, pH, ionic strength, and wet/dry beads were investigated. The pseudo first-order, second-order kinetic models and intraparticle diffusion model were used to describe the kinetic data and the rate constants were evaluated. The dynamical data fit well with the second-order kinetic model, except for the dry beads fitting better with the first-order model. The adsorption capacity increases largely with decreasing solution pH or with increasing initial dye concentration. Thermodynamic parameters such as change in free energy (deltaG(0)), enthalpy (deltaH(0)), entropy (deltaS(0)) and activation energy were also determined. The adsorption mechanism is shown to be the electrostatic interactions between the dye and chitosan beads. The desorption data shows that the removal percent of dye RR 189 from the cross-linked chitosan beads is 63% in NaOH solutions at pH 10.0, 30 degrees C. The desorbed chitosan beads can be reused to adsorb the dye and to reach the same capacity as that before desorption. PMID- 12531718 TI - Sonochemically induced decomposition of energetic materials in aqueous media. AB - This study demonstrates that ultrasound rapidly degrades the energetic compounds RDX (cyclo-1,3,5-trinitramine-2,4,6-trimethylene) and ADN (ammonium dinitramide) in aqueous microheterogeneous media. The conditions for effective degradation of these nitramines, as monitored by UV absorption spectroscopy, were determined by varying sonication time, the heterogeneous phase and its suspension density, and the concentration of NaOH. In the presence of 5 mg/ml of aluminum powder and at pH approximately 12 (10 mM NaOH), 74% of the RDX and 86% of the ammonium dinitramide (ADN) in near-saturated solutions decompose within the first 20 min of sonication (20 kHz; 50 W; < or =5 degrees C). Sonication without Al powder and base yields minimal degradation of either RDX and ADN (approximately 5-10%) or the nitrite/nitrate ions that are expected byproducts during RDX and ADN degradation. Sonication at high pH in the presence of dispersed aluminosilicate zeolite, alumina, or titanium dioxide also yields minimal degradation. Preliminary electrochemical studies and product analyses indicate that in situ ultrasonic generation of metallic aluminum and/or aluminum hydride drives reductive denitration of the nitramines. Sonochemical treatment in the presence of a reductant offers an effective and rapid waste remediation option for energetic waste compounds. PMID- 12531719 TI - Characterization and reactivity of natural manganese ore catalysts in the selective catalytic oxidation of ammonia to nitrogen. AB - Natural manganese ore (NMO) catalysts were characterized and tested in the selective catalytic oxidation of ammonia to nitrogen oxides under dilute conditions. Also, the oxidation of ammonia (NH(3)) was carried out using pure MnO(2), Mn(2)O(3) for comparing with the activity. It is found that the activity of NMO was similar to that of MnO(2) at low temperature below 150 degrees C but above this temperature, the activity of these catalysts showed the difference. In the course of NH(3) oxidation, N(2), NO, N(2)O and H(2)O were produced. But the quantity of NO(2) produced in this experiment was negligible. At temperature below 250 degrees C, selectivity into N(2) from NH(3) oxidation was in the order, NMO > MnO(2) > Mn(2)O(3). This is the reverse of activity of these manganese oxides. Also the characterization of NH(3) oxidation was proposed and supported by the effect of space velocity, inlet O(2) and NH(3) concentration. The increase of space velocity remarkably influenced not only the conversion but also selectivity into N(2). The higher the reaction temperature was, the higher the effect of inlet O(2) and NH(3) concentration on the reaction rate was. By introducing NO during NH(3) oxidation reaction, the possibility of NMO as selective catalytic reduction catalyst at low temperature was studied and showed positive results. PMID- 12531720 TI - Degradation of alpha-actin filaments in venous smooth muscle cells in response to mechanical stretch. AB - Mechanical stretch has been shown to induce the degradation of alpha-actin filaments in smooth muscle cells (SMC) of experimental vein grafts. Here, we investigate the possible role of ERK1/2 and p38 MAPK in regulating this process using an ex vivo venous culture model that simulates an experimental vein graft. An exposure of a vein to arterial pressure induced a significant increase in the medial circumferential strain, which induced rapid alpha-actin filament disruption, followed by degradation. The percentage of SMC alpha-actin filament coverage was reduced significantly under arterial pressure (91 +/- 1%, 43 +/- 13%, 51 +/- 5%, 28 +/- 3%, and 19 +/- 5% at 1, 6, 12, 24, and 48 h, respectively), whereas it did not change significantly in specimens under venous pressure at theses times. The degradation of SMC alpha-actin filaments paralleled an increase in the relative activity of caspase 3 (3.0 +/- 0.7- and 1.7 +/- 0.4 fold increase relative to the control level at 6 and 12 h, respectively) and a decrease in SMC density (from the control level of 1,368 +/- 66 cells/mm(2) at time 0 to 1,205 +/- 90, 783 +/- 129, 845 +/- 61, 637 +/- 55, and 432 +/- 125 cells/mm(2) at 1, 6, 12, 24, and 48 h of exposure to arterial pressure, respectively). Treatment with a p38 MAPK inhibitor (SB-203580) significantly reduced the stretch-induced activation of caspase 3 at 6 h (from 3.0 +/- 0.7- to 2.2 +/- 0.3-fold) in conjunction with a significant rescue of alpha-actin filament degradation (from 43 +/- 13% to 69 +/- 15%) at the same time. Treatment with an inhibitor for the ERK1/2 activator (PD-98059), however, did not induce a significant change in the activity of caspase 3 or the percentage of SMC alpha actin filament coverage. These results suggest that p38 MAPK and caspase 3 may mediate stretch-dependent degradation of alpha-actin filaments in vascular SMCs. PMID- 12531721 TI - Branching tree model with fractal vascular resistance explains fractal perfusion heterogeneity. AB - Perfusion heterogeneities in organs such as the heart obey a power law as a function of scale, a behavior termed "fractal." An explanation of why vascular systems produce such a specific perfusion pattern is still lacking. An intuitive branching tree model is presented that reveals how this behavior can be generated as a consequence of scale-independent branching asymmetry and fractal vessel resistance. Comparison of computer simulations to experimental data from the sheep heart shows that the values of the two free model parameters are realistic. Branching asymmetry within the model is defined by the relative tissue volume being fed by each branch. Vessel ordering for fractal analysis of morphology based on fed or drained tissue volumes is preferable to the commonly used Strahler system, which is shown to depend on branching asymmetry. Recently, noninvasive imaging techniques such as PET and MRI have been used to measure perfusion heterogeneity. The model allows a physiological interpretation of the measured fractal parameters, which could in turn be used to characterize vascular morphology and function. PMID- 12531722 TI - Oxytocinergic regulation of cardiovascular function: studies in oxytocin deficient mice. AB - Oxytocin (OT) has been implicated in the cardiovascular responses to exercise, stress, and baroreflex adjustments. Studies were conducted to determine the effect of genetic manipulation of the OT gene on blood pressure (BP), heart rate (HR), and autonomic/baroreflex function. OT knockout (OTKO -/-) and control +/+ mice were prepared with chronic arterial catheters. OTKO -/- mice exhibited a mild hypotension (102 +/- 3 vs. 110 +/- 3 mmHg). Sympathetic and vagal tone were tested using beta(1)-adrenergic and cholinergic blockade (atenolol and atropine). Magnitude of sympathetic and vagal tone to the heart and periphery was not significantly different between groups. However, there was an upward shift of sympathetic tone to higher HR values in OTKO -/- mice. This displacement combined with unchanged basal HR led to larger responses to cholinergic blockade (+77 +/- 25 vs. +5 +/- 15 beats/min, OTKO -/- vs. control +/+ group). There was also an increase in baroreflex gain (-13.1 +/- 2.5 vs. -4.1 +/- 1.2 beats x min(-1) x mmHg(-1), OTKO -/- vs. control +/+ group) over a smaller BP range. Results show that OTKO -/- mice are characterized by 1) hypotension, suggesting that OT is involved in tonic BP maintenance; 2) enhanced baroreflex gain over a small BP range, suggesting that OT extends the functional range of arterial baroreceptor reflex; and 3) shift in autonomic balance, indicating that OT reduces the sympathetic reserve. PMID- 12531723 TI - Impact of estrogen replacement on ventricular myocyte contractile function and protein kinase B/Akt activation. AB - Women with functional ovaries have a lower cardiovascular risk than men and postmenopausal women. However, estrogen replacement therapy remains controversial. This study examined the effect of ovarian hormone deficiency and estrogen replacement on ventricular myocyte contractile function and PKB/Akt activation. Nulliparous female rats were subjected to bilateral ovariectomy (Ovx) or sham operation (sham). A subgroup of Ovx rats received estrogen (E(2)) replacement (40 microg. kg(-1). day(-1)) for 8 weeks. Mechanical and intracellular Ca(2+) properties were evaluated including peak shortening (PS), time to PS (TPS), time to 90% relengthening (TR(90)), maximal velocity of shortening/relengthening (+/-dL/dt), fura 2 fluorescence intensity (FFI), and decay rate. Levels of sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA2a), phospholamban (PLB), and Akt were assessed by Western blot. Ovx promoted body weight gain associated with reduced serum E(2) and uterine weight, all of which were abolished by E(2). Ovx depressed PS and +/-dL/dt, prolonged TPS, TR(90), and decay rate, and enhanced resting FFI, all of which, with the exception of TPS, were restored by E(2). Ovx did not alter the levels of SERCA2a, PLB, and total Akt, but significantly reduced Akt activation [phosphorylated Akt (pAkt)], pAkt/Akt, and the SERCA2a-to-PLB ratio. These alterations in protein expression were restored by E(2). E(2) enhanced PS and +dL/dt in vitro, which was abolished by the E(2) receptor antagonist ICI-182780. Ovx reduced myocyte Ca(2+) responsiveness and lessened stimulating frequency-induced decline in PS, both ablated by E(2). These data suggest that mechanical and protein functions of ventricular myocytes are directly regulated by E(2). PMID- 12531724 TI - Central interleukin-1beta antibody increases renal and splenic sympathetic nerve discharge. AB - We tested the hypothesis that intracerebroventricular (lateral ventricle) administration of interleukin-1beta (IL-1beta) antibody increases the level of sympathetic nerve discharge (SND) in alpha-chloralose-anesthetized rats. Mean arterial pressure (MAP), heart rate (HR), and SND (splenic and renal) were recorded before (Preinfusion), during (25 min), and for 45 min after infusion of IL-1beta antibody (15 microg, 50 microl icv) in baroreceptor-intact (intact) and sinoaortic-denervated (SAD) rats. The following observations were made. First, intracerebroventricular infusion of IL-1beta antibody (but not saline and IgG) significantly increased MAP and the pressor response was higher in SAD compared with intact rats. Second, renal and splenic SND were significantly increased during and after intracerebroventricular IL-1beta antibody infusion and sympathoexcitatory responses were higher in SAD compared with intact rats. Third, intracerebroventricular administration of a single dose of IL-1beta antibody (15 microg, 5 microl for 2 min) significantly increased splenic and renal SND in intact rats. These results suggest that under the conditions of the present experiments central neural IL-1beta plays a role in the tonic regulation of SND and arterial blood pressure. PMID- 12531725 TI - Multitrack system for superfusing isolated cardiac myocytes. AB - A new system for studying mechanical activity of freshly isolated cardiac myocytes from up to four experimental groups simultaneously is described. Suspensions of cardiac myocytes isolated from adult rat hearts were drawn into microhematocrit capillary tubes, which were then mounted in parallel fashion between two four-channel tubing manifolds placed on the movable stage of an inverted microscope. Within a few minutes, cells settled and attached to the bottom of the tubes and then could be superfused with various test solutions. The system allowed for electrical field stimulation, rapid changes in bathing solutions, control of temperature, and simulation of ischemia and reperfusion with measurements of the effects of such interventions on both populations of cells (low power survey) and individual myocytes (high power). Myocyte responses to these various interventions are described. The primary advantage of this system is the ability to conduct experiments on cardiac myocytes isolated concurrently from multiple experimental groups at the same time and under identical conditions. PMID- 12531726 TI - Fibroblast alignment under interstitial fluid flow using a novel 3-D tissue culture model. AB - Interstitial flow is an important component of the microcirculation and interstitial environment, yet its effects on cell organization and tissue architecture are poorly understood, in part due to the lack of in vitro models. To examine the effects of interstitial flow on cell morphology and matrix remodeling, we developed a tissue culture model that physically supports soft tissue cultures and allows microscopic visualization of cells within the three dimensional matrix. In addition, pressure-flow relationships can be continuously monitored to evaluate the bulk hydraulic resistance as an indicator of changes in the overall matrix integrity. We observed that cells such as human dermal fibroblasts aligned perpendicular to the direction of interstitial flow. In contrast, fibroblasts in static three-dimensional controls remained randomly oriented, whereas cells subjected to fluid shear as a two-dimensional monolayer regressed. Also, the dynamic measurements of hydraulic conductivity suggest reorganization toward a steady state. These primary findings help establish the importance of interstitial flow on the biology of tissue organization and interstitial fluid balance. PMID- 12531727 TI - Single-beat estimation of right ventricular end-systolic pressure-volume relationship. AB - Assessment of right ventricular (RV) contractility from end-systolic pressure volume relationships (ESPVR) is difficult due to problems in measuring RV instantaneous volume and to effects of changes in RV preload or afterload. We therefore investigated in anesthetized dogs whether RV ESPVR and contractility can be determined without measuring RV volume and without changing RV preload or afterload. The maximal RV pressure of isovolumic beats (P(max)) was predicted from isovolumic portions of RV pressure during ejecting beats and compared with P(max) measured during the first beat after pulmonary artery clamping. In RV pressure-volume loops obtained from RV pressure and integrated pulmonary arterial flow, end-systolic elastance (E(es)) was assessed as the slope of P(max)-derived ESPVR, pulmonary artery effective elastance (E(a)) as the slope of end-diastolic to end-systolic relation, and coupling efficiency as the E(es)-to-E(a) ratio (E(es)/E(a)). Predicted P(max) correlated with observed P(max) (r = 0.98 +/- 0.02). Dobutamine increased E(es) from 1.07 to 2.00 mmHg/ml and E(es)/E(a) from 1.64 to 2.49, and propranolol decreased E(es)/E(a) from 1.64 to 0.91 (all P < 0.05). After adrenergic blockade, preload reduction did not affect E(es), whereas hypoxia and arterial constriction markedly increased E(a) and somewhat increased E(es) due to the Anrep effect. Low preload did not affect E(es)/E(a) and high afterload decreased E(es)/E(a). In conclusion, in the right ventricle 1) P(max) can be calculated from normal beats, 2) P(max) can be used to determine ESPVR without change in load, and 3) P(max)-derived ESPVR can be used to assess ventricular contractility and ventricular-arterial coupling efficiency. PMID- 12531728 TI - Glucocorticoids reduce responses to AMPA receptor activation and blockade in nucleus tractus solitarius. AB - We tested the hypothesis that glucocorticoids attenuate changes in arterial pressure and renal sympathetic nerve activity (RSNA) in response to activation and blockade of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors within the nucleus of the solitary tract (NTS). Experiments were performed in Inactin-anesthetized male Sprague-Dawley rats treated for 7 +/- 1 days with a subcutaneous corticosterone (Cort) pellet or in control rats. Baseline mean arterial pressure (MAP) was significantly higher in Cort-treated rats (109 +/- 2 mmHg, n = 39) than in control rats (101 +/- 1 mmHg, n = 48, P < 0.05). In control rats, microinjection of AMPA (0.03, 0.1, and 0.3 pmol/100 nl) into the NTS significantly decreased MAP at all doses and decreased RSNA at 0.1 and 0.3 pmol/100 nl. Responses to AMPA in Cort-treated rats were attenuated at all doses of AMPA (P < 0.05). Responses to the AMPA-kainate receptor antagonist 6 cyano-7-nitroquinoxaline-2,3-dione (CNQX) were also significantly reduced in Cort treated rats relative to control rats. Blockade of glucocorticoid type II receptors with mifepristone significantly enhanced responses to CNQX in both control and Cort rats. We conclude that glucocorticoids attenuate MAP and RSNA responses to activation and blockade of AMPA receptors in the NTS. PMID- 12531729 TI - Time-domain representation of ventricular-arterial coupling as a windkessel and wave system. AB - The differences in shape between central aortic pressure (P(Ao)) and flow waveforms have never been explained satisfactorily in that the assumed explanation (substantial reflected waves during diastole) remains controversial. As an alternative to the widely accepted frequency-domain model of arterial hemodynamics, we propose a functional, time-domain, arterial model that combines a blood conducting system and a reservoir (i.e., Frank's hydraulic integrator, the windkessel). In 15 anesthetized dogs, we measured P(Ao), flows, and dimensions and calculated windkessel pressure (P(Wk)) and volume (V(Wk)). We found that P(Wk) is proportional to thoracic aortic volume and that the volume of the thoracic aorta comprises 45.1 +/- 2.0% (mean +/- SE) of the total V(Wk). When we subtracted P(Wk) from P(Ao), we found that the difference (excess pressure) was proportional to aortic flow, thus resolving the differences between P(Ao) and flow waveforms and implying that reflected waves were minimal. We suggest that P(Ao) is the instantaneous summation of a time-varying reservoir pressure (i.e., P(Wk)) and the effects of (primarily) forward-traveling waves in this animal model. PMID- 12531730 TI - Cysteinyl leukotriene-dependent [Ca2+]i responses to angiotensin II in cardiomyocytes. AB - With the use of fura 2 measurements in multiple and single cells, we examined whether cysteinyl leukotrienes (CysLT) mediate angiotensin II (ANG II)-evoked increases in cytosolic free Ca(2+) concentration ([Ca(2+)](i)) in neonatal rat cardiomyocytes. ANG II-evoked CysLT release peaked at 1 min. The angiotensin type 1 (AT(1)) antagonist losartan, but not the AT(2) antagonist PD-123319, attenuated the elevations in [Ca(2+)](i) and CysLT levels evoked by ANG II. Vasopressin and endothelin-1 increased [Ca(2+)](i) but not CysLT levels. The 5-lipoxygenase (5 LO) inhibitor AA-861 and the CysLT(1)-selective antagonist MK-571 reduced the maximal [Ca(2+)](i) responses to ANG II but not to vasopressin and endothelin-1. While MK-571 reduced the responses to leukotriene D(4) (LTD(4)), the dual CysLT antagonist BAY-u9773 completely blocked the [Ca(2+)](i) elevation to both LTD(4) and LTC(4). These data confirm that ANG II-evoked increases, but not vasopressin- and endothelin-1-evoked increases, in [Ca(2+)](i) involve generation of the 5 lipoxygenase metabolite CysLT. The inositol (1,4,5)-trisphosphate [Ins(1,4,5)P(3)] antagonist 2-aminoethoxydiphenyl borate attenuated the [Ca(2+)](i) responses to ANG II and LTD(4). Thus AT(1) receptor activation by ANG II is linked to CysLT-mediated Ca(2+) release from Ins(1,4,5)P(3)-sensitive intracellular stores to augment direct ANG II-evoked Ca(2+) mobilization in rat cardiomyocytes. PMID- 12531731 TI - An electrodiffusion model for effects of surface glycocalyx layer on microvessel permeability. AB - To investigate the charge effect of the endothelial surface glycocalyx on microvessel permeability, we extended the three-dimensional model developed by Fu et al. (J Biomech Eng 116: 502-513, 1994) for the interendothelial cleft to include a negatively charged glycocalyx layer at the entrance of the cleft. Both electrostatic and steric exclusions on charged solutes were considered within the glycocalyx layer and at the interfaces. Four charge-density profiles were assumed for the glycocalyx layer. Our model indicates that the overall solute permeability across the microvessel wall including the surface glycocalyx layer and the cleft region is independent of the charge-density profiles as long as they have the same maximum value and the same total charge. On the basis of experimental data, this model predicts that the charge density would be 25-35 meq/l in the glycolcalyx of frog mesenteric capillaries. An intriguing prediction of this model is that when the concentrations of cations and anions are unequal in the lumen due to the presence of negatively charged proteins, the negatively charged glycocalyx would provide more resistance to positively charged solutes than to negatively charged ones. PMID- 12531732 TI - Age-dependent increase in hydrogen peroxide production by cardiac monoamine oxidase A in rats. AB - Oxidative stress is one of the factors involved in age-related impairment of cardiac function. In the present study, we investigated the role of the catecholamine-degrading enzyme monoamine oxidase (MAO) in H(2)O(2) production in the hearts of young, adult, and old rats. MAO-dependent H(2)O(2) production, measured by a chemiluminescence-based assay, increased with age, reaching the maximum in 24-mo-old rats (7.5-fold increase vs. 1-mo-old rats). The following observations indicate that the age-dependent increase in H(2)O(2) generation was fully related to the MAO-A isoform: 1) at all the ages tested, chemiluminescence production was inhibited by the MAO-A inhibitor clorgyline but not by the MAO-B inhibitor RO-19 6327; 2) enzyme assay, Western blot, and semiquantitative RT-PCR analysis showed an age-dependent increase in cardiac MAO-A activity, immunodetection, and mRNA expression, respectively; and 3) the MAO-B isoform was undetectable by enzyme assay and Western blot analysis. These results suggest that MAO-A could be a major source of H(2)O(2) in the aging heart. PMID- 12531733 TI - Epitope-dependent localization of estrogen receptor-alpha, but not -beta, in en face arterial endothelium. AB - Rapid, nongenomic effects of 17 beta-estradiol (E(2)) in endothelial cells are postulated to arise from membrane-associated estrogen receptors (ERs), which have not been visualized in vascular tissue. To identify membrane ERs, we used multiple site-directed ER alpha or ER beta antibodies to label en face rat cerebral and coronary arterial endothelia. Western blots revealed a novel 55-kDa ER alpha isoform. Three-dimensional images of cells labeled with these antibodies and markers for the nucleus and caveolin-1 were acquired with a wide-field microscope, deconvolved, and numerically analyzed. We found ER alpha in the nucleus and cell periphery, where one-third colocalized with caveolin-1. The receptor location was dependent on the epitope of the antibody. Human ovarian surface epithelium produced similar results; but in rat myometrium, the distribution was epitope independent and nuclear. ER beta distribution was predominantly intranuclear and epitope independent. A small amount of ER alpha colocalized with ER beta within the nucleus. The results were identical in both arterial preparations and insensitive to E(2). We postulate that the different ER alpha conformations at the membrane, in the nucleus, and between different cell types allow E(2) to trigger cell- and location-specific signaling cascades. PMID- 12531734 TI - Protein tyrosine kinase signaling is necessary for NO donor-induced late preconditioning against myocardial stunning. AB - Although protein tyrosine kinases (PTKs) signaling has been implicated in the late phase of ischemic preconditioning (PC), it is unknown whether PTK signaling is necessary for the development of nitric oxide (NO) donor-induced late PC. Thus conscious rabbits underwent a sequence of six 4-min coronary occlusion (O)/4-min reperfusion (R) cycles followed by a 5-h recovery period of reperfusion for 3 consecutive days (days 1, 2, and 3). On day 0 (24 h before the 6 O/R cycles on day 1), rabbits received no treatment (control), the NO donor diethylenetriamine (DETA)/NO (DETA/NO), the PTK inhibitor 4-amino-5-(4-chlorophenyl)-7-(t butyl)pyrazolo[3,4-d]pyrimidine (PP2), or DETA/NO plus PP2 (DETA/NO + PP2). In control rabbits (n = 6), the six O/R cycles on day 1 resulted in delayed functional recovery, indicating severe myocardial stunning. In rabbits pretreated with DETA/NO (n = 5) on day 1, myocardial stunning caused by the six O/R cycles on day 1 was markedly attenuated, with a significant reduction ( approximately 60%) in the total deficit of wall thickening (WTh) compared with controls, indicating that DETA/NO induced a late PC effect against stunning. However, in rabbits pretreated with DETA/NO + PP2 (n = 5), the total deficit of WTh was significantly greater than that in rabbits treated with DETA/NO alone and was similar to that in controls, indicating that PP2 prevented the development of DETA/NO-induced late PC. In rabbits pretreated with PP2 on day 0 (n = 4), the total deficit of WTh was similar to that in controls, indicating that PP2 does not affect myocardial stunning in itself. We conclude that a PTK-dependent signaling mechanism is necessary for the development of NO donor-induced late PC against myocardial stunning in conscious rabbits. PMID- 12531735 TI - Cross-bridge kinetics modeled from myoplasmic [Ca2+] and LV pressure at 17 degrees C and after 37 degrees C and 17 degrees C ischemia. AB - We modeled changes in contractile element kinetics derived from the cyclic relationship between myoplasmic [Ca(2+)], measured by indo 1 fluorescence, and left ventricular pressure (LVP). We estimated model rate constants of the Ca(2+) affinity for troponin C (TnC) on actin (A) filament (TnCA) and actin and myosin (M) cross-bridge (A x M) cycling in intact guinea pig hearts during baseline 37 degrees C perfusion and evaluated changes at 1) 20 min 17 degrees C pressure, 2) 30-min reperfusion (RP) after 30-min 37 degrees C global ischemia during 37 degrees C RP, and 3) 30-min RP after 240-min 17 degrees C global ischemia during 37 degrees C RP. At 17 degrees C perfusion versus 37 degrees C perfusion, the model predicted: A x M binding was less sensitive; A x M dissociation was slower; Ca(2+) was less likely to bind to TnCA with A x M present; and Ca(2+) and TnCA binding was less sensitive in the absence of A x M. Model results were consistent with a cold-induced fall in heart rate from 260 beats/min (37 degrees C) to 33 beats/min (17 degrees C), increased diastolic LVP, and increased phasic Ca(2+). On RP after 37 degrees C ischemia vs. 37 degrees C perfusion, the model predicted the following: A x M binding was less sensitive; A x M dissociation was slower; and Ca(2+) was less likely to bind to TnCA in the absence of A. M. Model results were consistent with reduced myofilament responsiveness to [Ca(2+)] and diastolic contracture on 37 degrees C RP. In contrast, after cold ischemia versus 37 degrees C perfusion, A x M association and dissociation rates, and Ca(2+) and TnCA association rates, returned to preischemic values, whereas the dissociation rate of Ca(2+) from A x M was ninefold faster. This cardiac muscle kinetic model predicted a better-restored relationship between Ca(2+) and cross-bridge function on RP after an eightfold longer period of 17 degrees C than 37 degrees C ischemia. PMID- 12531736 TI - Effect of cortisol on norepinephrine-mediated contractions in ovine uterine arteries. AB - Cortisol potentiated norepinephrine (NE)-mediated contractions in ovine uterine arteries (UA). We tested the hypothesis that cortisol regulated alpha(1) adrenoceptor-mediated pharmacomechanical coupling differentially in nonpregnant UA (NUA) and pregnant UA (PUA). Cortisol (10 ng/ml for 24 h) significantly increased contractile coupling efficiency of alpha(1)-adrenoceptors in NUA, but increased alpha(1)-adrenoceptor density in PUA. Cortisol potentiated NE-induced inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)] synthesis in both NUA and PUA, but increased coupling efficiency of alpha(1)-adrenoceptors to Ins(1,4,5)P(3) synthesis only in NUA. Carbenoxolone alone did not affect NE-mediated Ins(1,4,5)P(3) production, but significantly enhanced cortisol-mediated potentiation of NE-stimulated Ins(1,4,5)P(3) synthesis in PUA. In addition, cortisol potentiated the NE-induced increase in Ca(2+) concentration in PUA, but increased NE-mediated contraction for a given amount of Ca(2+) concentration in NUA. Collectively, the results indicate that cortisol potentiates NE-mediated contractions differentially in NUA and PUA, i.e., by upregulating alpha(1) adrenoceptor density leading to increased Ca(2+) mobilization in PUA while increasing alpha(1)-adrenoceptor coupling efficiency and myofilament Ca(2+) sensitivity in NUA. In addition, the results suggest that pregnancy increases type 2 11 beta-hydroxysteroid dehydrogenase activity in the UA. PMID- 12531737 TI - A mathematical model of phase 2 reentry: role of L-type Ca current. AB - Phase 2 reentry (P2R) is known to be one of the mechanisms of malignant ventricular arrhythmias, especially those associated with Brugada syndrome. However, little is known about the underlying mechanism for P2R. Our aim in this study was to simulate P2R in a mathematical model to enable us to understand its mechanism and identify a potential therapeutic target. A mathematical model of the L-type Ca current was composed according to whole cell current data from guinea pig ventricular myocytes recorded at 37 degrees C. Our mathematical model was incorporated into the modified Luo-Rudy phase 2 model. We set a dispersion in transient outward current (I(to)) density within the theoretical fiber, composed of 80 serially arranged epicardial cells with gap junctions and then observed the P2R. The dispersion in I(to) density within an only 0.8-cm epicardial theoretical fiber generated P2R with our Ca channel but not with the original model. When the P2R developed in the theoretical fiber, the calculated extracellular field potential showed coved-type ST segment elevation. We succeeded in generating P2R in our model for the first time. The local epicardial P2R may contribute the genesis of coved-type ST segment elevation in the Brugada syndrome. PMID- 12531738 TI - Cardiovascular effects of hypocretin-1 in nucleus of the solitary tract. AB - Experiments were done in male Wistar rats to investigate the effects of microinjection of hypocretin-1 (Hcrt-1) into the nucleus of the solitary tract (NTS) on mean arterial pressure (MAP), heart rate (HR), and the baroreflex. In the first series, the distribution of Hcrt-1-like immunoreactivity (Ir) was mapped within the region of NTS. Hcrt-1 Ir was found throughout the NTS region, predominantly within the caudal dorsolateral (Slt), medial (Sm), and interstitial subnuclei of the NTS. In the second series, in alpha-chloralose or urethane anesthetized rats, microinjection of Hcrt-1 (0.5-5 pmol) into the caudal NTS elicited a dose-dependent decrease in MAP and HR. A mapping of the caudal NTS region showed that the largest depressor and bradycardia responses elicited by Hcrt-1 were from sites in the Slt and Sm. In addition, doses >2.5 pmol at a small number of sites localized to the caudal commissural nucleus of NTS elicited pressor and tachycardia responses. Intravenous administration of the muscarinic receptor blocker atropine methyl bromide abolished the bradycardia response and attenuated the depressor response, whereas subsequent administration of the nicotinic receptor blocker hexamethonium bromide abolished the remaining MAP response. Finally, microinjection of Hcrt-1 into the NTS significantly potentiated the reflex bradycardia to activation of arterial baroreceptors as a result of increasing MAP by systemic injections of phenylephrine (2-4 microg/kg). These results suggest that Hcrt-1 in the NTS activates neuronal circuits that increases vagal activity to the heart, inhibits sympathetic activity to the heart and vasculature, and alters the excitability of NTS neuronal circuits that reflexly control the circulation. PMID- 12531739 TI - Blood flow and muscle metabolism: a focus on insulin action. AB - The vascular system controls the delivery of nutrients and hormones to muscle, and a number of hormones may act to regulate muscle metabolism and contractile performance by modulating blood flow to and within muscle. This review examines evidence that insulin has major hemodynamic effects to influence muscle metabolism. Whole body, isolated hindlimb perfusion studies and experiments with cell cultures suggest that the hemodynamic effects of insulin emanate from the vasculature itself and involve nitric oxide-dependent vasodilation at large and small vessels with the purpose of increasing access for insulin and nutrients to the interstitium and muscle cells. Recently developed techniques for detecting changes in microvascular flow, specifically capillary recruitment in muscle, indicate this to be a key site for early insulin action at physiological levels in rats and humans. In the absence of increases in bulk flow to muscle, insulin may act to switch flow from nonnutritive to the nutritive route. In addition, there is accumulating evidence to suggest that insulin resistance of muscle in vivo in terms of impaired glucose uptake could be partly due to impaired insulin mediated capillary recruitment. Exercise training improves insulin-mediated capillary recruitment and glucose uptake by muscle. PMID- 12531740 TI - Stem/progenitor cells derived from adult tissues: potential for the treatment of diabetes mellitus. AB - In view of the recent success in pancreatic islet transplantation, interest in treating diabetes by the delivery of insulin-producing beta-cells has been renewed. Because differentiated pancreatic beta-cells cannot be expanded significantly in vitro, beta-cell stem or progenitor cells are seen as a potential source for the preparation of transplantable insulin-producing tissue. In addition to embryonic stem (ES) cells, several potential adult islet/beta-cell progenitors, derived from pancreas, liver, and bone marrow, are being studied. To date, none of the candidate cells has been fully characterized or is clinically applicable, but pancreatic physiology makes the existence of one or more types of adult islet stem cells very likely. It also seems possible that pluripotential stem cells, derived from the bone marrow, contribute to adult islet neogenesis. In future studies, more stringent criteria should be met to clonally define adult islet/beta-cell progenitor cells. If this can be achieved, the utilization of these cells for the generation of insulin-producing beta-cells in vitro seems to be feasible in the near future. PMID- 12531741 TI - Magnitude and variation of fat-free mass density: a cellular-level body composition modeling study. AB - The mean density of fat-free mass (FFM) is remarkably stable at 1.10 g/cm(3) in healthy adult humans, and this stability is a cornerstone of the widely applied densitometry-based two-compartment model for estimating total body fat. At present, the usual means of exploring FFM density is by in vitro or in vivo experimental studies. The purpose of the present investigation was to develop a cellular-level body composition model that includes seven factors that determine FFM density. The model, when applied with available empirical coefficients, predicted an FFM density similar to that observed in vivo. An analysis of the seven model components indicates that the ratio of extracellular solids to total body water is a major determinant of individual variation in FFM density. The difference in FFM density across sex, race, and age groups was examined with the developed model. The present study thus provides a conceptual framework for the systematic study of FFM density in humans. PMID- 12531742 TI - Free fatty acids increase basal hepatic glucose production and induce hepatic insulin resistance at different sites. AB - To investigate the sites of the free fatty acid (FFA) effects to increase basal hepatic glucose production and to impair hepatic insulin action, we performed 2-h and 7-h Intralipid + heparin (IH) and saline infusions in the basal fasting state and during hyperinsulinemic clamps in overnight-fasted rats. We measured endogenous glucose production (EGP), total glucose output (TGO, the flux through glucose-6-phosphatase), glucose cycling (GC, index of flux through glucokinase = TGO - EGP), hepatic glucose 6-phosphate (G-6-P) content, and hepatic glucose-6 phosphatase and glucokinase activities. Plasma FFA levels were elevated about threefold by IH. In the basal state, IH increased TGO, in vivo glucose-6 phosphatase activity (TGO/G-6-P), and EGP (P < 0.001). During the clamp compared with the basal experiments, 2-h insulin infusion increased GC and in vivo glucokinase activity (GC/TGO; P < 0.05) and suppressed EGP (P < 0.05) but failed to significantly affect TGO and in vivo glucose-6-phosphatase activity. IH decreased the ability of insulin to increase GC and in vivo glucokinase activity (P < 0.01), and at 7 h, it also decreased the ability of insulin to suppress EGP (P < 0.001). G-6-P content was comparable in all groups. In vivo glucose-6 phosphatase and glucokinase activities did not correspond to their in vitro activities as determined in liver tissue, suggesting that stable changes in enzyme activity were not responsible for the FFA effects. The data suggest that, in overnight-fasted rats, FFA increased basal EGP and induced hepatic insulin resistance at different sites. 1) FFA increased basal EGP through an increase in TGO and in vivo glucose-6-phosphatase activity, presumably due to a stimulatory allosteric effect of fatty acyl-CoA on glucose-6-phosphatase. 2) FFA induced hepatic insulin resistance (decreased the ability of insulin to suppress EGP) through an impairment of insulin's ability to increase GC and in vivo glucokinase activity, presumably due to an inhibitory allosteric effect of fatty acyl-CoA on glucokinase and/or an impairment in glucokinase translocation. PMID- 12531743 TI - Interaction of free fatty acids and epinephrine in regulating hepatic glucose production in conscious dogs. AB - To determine the effects of an increase in lipolysis on the glycogenolytic effect of epinephrine (EPI), the catecholamine was infused portally into 18-h-fasted conscious dogs maintained on a pancreatic clamp in the presence [portal (Po) EPI+FFA, n = 6] and absence (Po-EPI+SAL, n = 6) of peripheral Intralipid infusion. Control groups with high glucose (70% increase) and free fatty acid (FFA; 200% increase; HG+FFA, n = 6) and high glucose alone (HG+SAL, n = 6) were also included. Hepatic sinusoidal EPI levels were elevated (Delta 568 +/- 77 and Delta 527 +/- 37 pg/ml, respectively) in Po-EPI+SAL and EPI+FFA but remained basal in HG+FFA and HG+SAL. Arterial plasma FFA increased from 613 +/- 73 to 1,633 +/- 101 and 746 +/- 112 to 1,898 +/- 237 micromol/l in Po-EPI+FFA and HG+FFA but did not change in EPI+SAL or HG+SAL. Net hepatic glycogenolysis increased from 1.5 +/- 0.3 to 3.1 +/- 0.4 mg x kg(-1) x min(-1) (P < 0.05) by 30 min in response to portal EPI but did not rise (1.8 +/- 0.2 to 2.1 +/- 0.3 mg x kg(-1) x min(-1)) in response to Po-EPI+FFA. Net hepatic glycogenolysis decreased from 1.7 +/- 0.2 to 0.9 +/- 0.2 and 1.6 +/- 0.2 to 0.7 +/- 0.2 mg x kg(-1) x min( 1) by 30 min in HG+FFA and HG+SAL. Hepatic gluconeogenic flux to glucose 6 phosphate increased from 0.6 +/- 0.1 to 1.2 +/- 0.1 mg x kg(-1) x min(-1) (P < 0.05; by 3 h) and 0.7 +/- 0.1 to 1.6 +/- 0.1 mg x kg(-1) x min(-1) (P < 0.05; at 90 min) in HG+FFA and Po-EPI+FFA. The gluconeogenic parameters remained unchanged in the Po-EPI+SAL and HG+SAL groups. In conclusion, increased FFA markedly changed the mechanism by which EPI stimulated hepatic glucose production, suggesting that its overall lipolytic effect may be important in determining its effect on the liver. PMID- 12531744 TI - The influence of insulin on circulating ghrelin. AB - Ghrelin is a novel peptide that acts on the growth hormone (GH) secretagogue receptor in the pituitary and hypothalamus. It may function as a third physiological regulator of GH secretion, along with GH-releasing hormone and somatostatin. In addition to the action of ghrelin on the GH axis, it appears to have a role in the determination of energy homeostasis. Although feeding suppresses ghrelin production and fasting stimulates ghrelin release, the underlying mechanisms controlling this process remain unclear. The purpose of this study was to test the hypotheses, by use of a stepped hyperinsulinemic eu- hypo- hyperglycemic glucose clamp, that either hyperinsulinemia or hypoglycemia may influence ghrelin production. Having been stable in the period before the clamp, ghrelin levels rapidly fell in response to insulin infusion during euglycemia (baseline ghrelin 207 +/- 12 vs. 169 +/- 10 fmol/ml at t = 30 min, P < 0.001). Ghrelin remained suppressed during subsequent periods of hypoglycemia (mean glucose 53 +/- 2 mg/dl) and hyperglycemia (mean glucose 163 +/- 6 mg/dl). Despite suppression of ghrelin, GH showed a significant rise during hypoglycemia (baseline 4.1 +/- 1.3 vs. 28.2 +/- 3.9 microg/l at t = 120 min, P < 0.001). Our data suggest that insulin may suppress circulating ghrelin independently of glucose, although glucose may have an additional effect. We conclude that the GH response seen during hypoglycemia is not regulated by circulating ghrelin. PMID- 12531745 TI - Impaired cardiac function and IGF-I response in myocytes from calmodulin-diabetic mice: role of Akt and RhoA. AB - This study characterized the cardiac contractile function and IGF-I response in a transgenic diabetic mouse model. Mechanical properties were evaluated in cardiac myocytes from OVE26 diabetic and FVB wild-type mice, including peak shortening (PS), time to PS (TPS), time to 90% relengthening (TR(90)) and maximal velocity of shortening/relengthening (+/-dL/dt). Intracellular Ca(2+) was evaluated as Ca(2+)-induced Ca(2+) release [difference in fura 2 fluorescent intensity (Delta FFI)] and fluorescence decay rate (tau). Sarco(endo)plasmic reticulum Ca(2+) ATPase (SERCA)2a, phospholamban (PLB), Na(+)-Ca(2+) exchanger (NCX), GLUT4, and the serine-threonine kinase Akt were assessed by Western blot. RhoA and IGF-I/IGF I receptor mRNA levels were determined by RT-PCR and Northern blot. OVE26 myocytes displayed decreased PS, +/-dL/dt, and Delta FFI associated with prolonged TPS, TR(90), and tau. SERCA2a, NCX, and Akt activation were reduced, whereas PLB and RhoA were enhanced in OVE26 hearts. GLUT4 was unchanged. IGF-I enhanced PS and Delta FFI in FVB but not OVE26 myocytes. IGF-I mRNA was increased, but IGF-I receptor mRNA was reduced in OVE26 hearts and livers. These results validate diabetic cardiomyopathy in OVE26 mice due to reduced SERCA2, NCX, IGF-I response, and Akt activation associated with enhanced RhoA level, suggesting a therapeutic potential for Akt and RhoA. PMID- 12531746 TI - Creatine transporter activity and content in the rat heart supplemented by and depleted of creatine. AB - The intracellular creatine concentration is an important bioenergetic parameter in cardiac muscle. Although creatine uptake is known to be via a NaCl-dependent creatine transporter (CrT), its localization and regulation are poorly understood. We investigated CrT kinetics in isolated perfused hearts and, by using cardiomyocytes, measured CrT content at the plasma membrane or in total lysates. Rats were fed control diet or diet supplemented with creatine or the creatine analog beta-guanidinopropionic acid (beta-GPA). Creatine transport in control hearts followed saturation kinetics with a K(m) of 70 +/- 13 mM and a V(max) of 3.7 +/- 0.07 nmol x min(-1) x g wet wt(-1). Creatine supplementation significantly decreased the V(max) of the CrT (2.7 +/- 0.17 nmol x min(-1) x g wet wt(-1)). This was matched by an approximately 35% decrease in the plasma membrane CrT; the total CrT pool was unchanged. Rats fed beta-GPA exhibited a >80% decrease in tissue creatine and increase in beta-GPA(total). The V(max) of the CrT was increased (6.0 +/- 0.25 nmol x min(-1) x g wet wt(-1)) and the K(m) decreased (39.8 +/- 3.0 mM). The plasma membrane CrT increased about fivefold, whereas the total CrT pool remained unchanged. We conclude that, in heart, creatine transport is determined by the content of a plasma membrane isoform of the CrT but not by the total cellular CrT pool. PMID- 12531747 TI - Potassium per kilogram fat-free mass and total body potassium: predictions from sex, age, and anthropometry. AB - Total body potassium (TBK) is located mainly intracellularly and constitutes an index of fat-free mass (FFM). The aim was to examine whether TBK and the TBK-to FFM ratio (TBK/FFM) can be estimated from sex, age, weight, and height. A primary study group (164 males, 205 females) and a validation group (161 and 206), aged 37-61 yr, were randomly selected from the general population. TBK was determined by whole body counting, and FFM was obtained by dual-energy X-ray absorptiometry (DEXA; FFM(DEXA)). The primary study group was used to construct sex-specific equations predicting TBK and TBK/FFM from age, weight, and height. The equations were used to estimate TBK and TBK/FFM in the validation group. The estimates were compared with measured values. TBK in different age ranges was predicted, with errors ranging from 5.0 to 6.8%; errors for TBK/FFM ranged from 2.7 to 4.8%, respectively. By adding FFM(DEXA) as a fourth predictor, the error of the TBK prediction decreased by approximately two percentage units. In conclusion, TBK and TBK/FFM can be meaningfully estimated from sex, age, weight, and height. PMID- 12531748 TI - Divergent effects of acute exercise and endurance training on UCP3 expression. PMID- 12531750 TI - Arthroscopic anterior cruciate ligament reconstruction: a metaanalysis comparing patellar tendon and hamstring tendon autografts. AB - BACKGROUND: The best choice of graft tissue for use in anterior cruciate ligament reconstruction has been the subject of debate. HYPOTHESIS: Anterior cruciate ligament reconstruction with patellar tendon autograft leads to greater knee stability than reconstruction with hamstring tendon autograft. STUDY DESIGN: Metaanalysis. METHODS: A Medline search identified articles published from January 1966 to May 2000 describing arthroscopic anterior cruciate ligament reconstruction with either patellar tendon or hamstring tendon autograft and with a minimum patient follow-up of 24 months. RESULTS: There were 1348 patients in the patellar tendon group (21 studies) and 628 patients in the hamstring tendon group (13 studies). The rate of graft failure in the patellar tendon group was significantly lower (1.9% versus 4.9%) and a significantly higher proportion of patients in the patellar tendon group had a side-to-side difference of less than 3 mm on KT-1000 arthrometer testing than in the hamstring tendon group (79% versus 73.8%). There was a higher rate of manipulation under anesthesia or lysis of adhesions (6.3% versus 3.3%) and of anterior knee pain in the patellar tendon group (17.4% versus 11.5%) and a higher incidence of hardware removal in the hamstring tendon group (5.5% versus 3.1%). CONCLUSIONS: Patellar tendon autografts had a significantly lower rate of graft failure and resulted in better static knee stability and increased patient satisfaction compared with hamstring tendon autografts. However, patellar tendon autograft reconstructions resulted in an increased rate of anterior knee pain. PMID- 12531751 TI - A prospective randomized study of patellar versus hamstring tendon autografts for anterior cruciate ligament reconstruction. AB - BACKGROUND: Bone-patellar tendon-bone graft has been the most commonly used graft material in anterior cruciate reconstructions, but there has been increasing use of hamstring tendon grafts. However, no existing clinical studies show adequate support for the choice of one graft over the other. HYPOTHESIS: Hamstring tendons are equally as good as patellar tendon in anterior cruciate ligament reconstructions. STUDY DESIGN: Prospective randomized clinical trial. METHODS: Ninety-nine patients with laxity caused by a torn anterior cruciate ligament underwent arthroscopically assisted reconstruction with graft randomization according to their birth year. Grafts were either bone-patellar tendon-bone with metal interference screw fixation or double-looped hamstring tendons with metal plate fixation. There were no significant differences between the two groups preoperatively or at operation. Standard rehabilitation included immediate postoperative mobilization without a knee brace, protected weightbearing for 2 weeks, and return to full activity at 6 to 12 months. RESULTS: Forty-three patients in the patellar tendon group and 46 patients in the hamstring tendon group were available for clinical evaluation at a minimum of 21 months after surgery. No statistically significant differences were seen with respect to clinical and instrumented laxity testing, International Knee Documentation Committee Score ratings, isokinetic muscle torque measurements, and Kujala patellofemoral, Lysholm, and Tegner scores. CONCLUSION: Equal results were seen for patellar and hamstring tendon autograft anterior cruciate ligament reconstructions at 2 years after surgery. Both techniques seem to improve patients' performance. PMID- 12531752 TI - Patellar tendon or semitendinosus tendon autografts for anterior cruciate ligament reconstruction? A prospective randomized study with a two-year follow up. AB - BACKGROUND: There are well-known problems with the use of bone-patellar tendon bone autografts for anterior cruciate ligament reconstruction, especially in terms of donor site morbidity. Hamstring tendon grafts have been increasingly used as an alternative, but there are very few controlled studies comparing the methods. HYPOTHESIS: Use of semitendinosus tendon grafts will cause less donor site morbidity and result in better knee-walking ability. STUDY DESIGN: Prospective randomized clinical trial. METHODS: Seventy-one patients who had a unilateral anterior cruciate ligament rupture underwent arthroscopic reconstruction with interference screw fixation and use of either bone-patellar tendon-bone or semitendinosus tendon graft. Outcome assessment was performed by physiotherapists not involved in the patients' care. RESULTS: At the 2-year follow-up, no differences were found in terms of the Lysholm score, Tegner activity level, KT-1000 arthrometer side-to-side laxity measurement, single legged hop test, or International Knee Documentation Committee classification results. The knee-walking test was rated difficult or impossible to perform by 53% of the bone-patellar tendon-bone group and by only 23% of the semitendinosus graft patients, a significant difference. CONCLUSIONS: The semitendinosus tendon graft is at least an equivalent option to the bone-patellar tendon-bone graft for anterior cruciate ligament reconstruction, and we recommend its use. PMID- 12531753 TI - Laser capsulorrhaphy for multidirectional instability of the shoulder. An outcomes study and proposed classification system. AB - BACKGROUND: Clinical data on the efficacy of laser capsulorrhaphy for the treatment of multidirectional instability of the shoulder are limited. HYPOTHESIS: The diagnosis of multidirectional instability includes a spectrum of pathologic symptoms that warrants subclassification; laser capsulorrhaphy alone is not uniformly effective for all subtypes. STUDY DESIGN: Retrospective review of prospectively collected data. METHODS: Twenty-five shoulders in 21 patients were treated with laser capsulorrhaphy for multidirectional instability. Functional outcomes at a mean duration of 32 months' follow-up (range, 24 to 48 months) were recorded. RESULTS: Instability recurred in 60% of patients with congenital multidirectional instability, 17% of patients with acquired multidirectional instability, and 33% of patients with posttraumatic multidirectional instability (overall recurrence rate, 40%). Generalized ligamentous laxity was a risk factor for recurrence. Patient satisfaction rates were 40%, 83%, and 22% for the congenital, acquired, and posttraumatic subgroups. Reasons for dissatisfaction included recurrent instability, persistent pain, and inability to return to athletic activity at desired capacity. The overall mean postoperative Simple Shoulder Test score was 84%. The mean postoperative numeric rating score for pain was 3.3 (10-point scale). CONCLUSIONS: Laser capsulorrhaphy may be effective for patients with acquired multidirectional instability secondary to repetitive microtrauma but is less predictable in the other subgroups. PMID- 12531754 TI - Influence of postoperative immobilization on tendon length after radiofrequency induced shrinkage. An in vivo rabbit study. AB - BACKGROUND: Despite the widespread use of radiofrequency-induced shrinkage of collagenous tissues, there have been no animal studies on the effects of postoperative immobilization after such treatment. PURPOSE: To examine the effects of postoperative immobilization after radiofrequency energy treatment, with special emphasis on any tissue length increases. STUDY DESIGN: Controlled laboratory study. METHODS: The right patellar tendon of 60 New Zealand White rabbits was shrunk with a radiofrequency probe. Tendon length was measured intraoperatively before and after shrinkage and via radiographs immediately postoperatively and at 3, 6, and 9 weeks. Twenty rabbits were not immobilized, 20 were immobilized for 3 weeks, and 20 were immobilized for 6 weeks. RESULTS: In the nonimmobilized limbs, the tendon length increased 34.9% at 3 weeks and another 2.5% at 6 weeks, versus 11.2% at 3 weeks and 6.6% at 6 weeks in the immobilized limbs. Ten of the 20 rabbits that were immobilized for 6 weeks were sacrificed at 9 weeks and were found to have a further length increase of 10.8%. At 9 weeks, the tendons of this group were no longer significantly shorter than the tendons from rabbits that had not been immobilized. CONCLUSIONS: Careful postoperative rehabilitation is imperative after radiofrequency-induced shrinkage. Without protection, exposure to normal physiologic loads places the shrunken tissue at risk of stretching out beyond the preshrinkage length. CLINICAL RELEVANCE: Shrunken tissue is at risk of stretching out after radiofrequency-induced shrinkage. PMID- 12531755 TI - Muscle flexibility as a risk factor for developing muscle injuries in male professional soccer players. A prospective study. AB - BACKGROUND: Muscular tightness is frequently postulated as an intrinsic risk factor for the development of a muscle injury. However, very little prospective data exist to prove this. HYPOTHESIS: Increased muscle tightness identifies a soccer player at risk for a subsequent musculoskeletal lesion. STUDY DESIGN: Prospective cohort study. METHODS: We examined 146 male professional soccer players before the 1999-2000 Belgian soccer competition. None of the players had a history of muscle injury in the lower extremities in the previous 2 years. The flexibility of the hamstring, quadriceps, adductor, and calf muscles of these players was measured goniometrically before the start of the season. All of the examined players were monitored throughout the season to register subsequent injuries. RESULTS: Players with a hamstring (N = 31) or quadriceps (N = 13) muscle injury were found to have significantly lower flexibility in these muscles before their injury compared with the uninjured group. No significant differences in muscle flexibility were found between players who sustained an adductor muscle injury (N = 13) or a calf muscle injury (N = 10) and the uninjured group. CONCLUSIONS: These results indicate that soccer players with an increased tightness of the hamstring or quadriceps muscles have a statistically higher risk for a subsequent musculoskeletal lesion. CLINICAL SIGNIFICANCE: Preseason hamstring and quadriceps muscle flexibility testing can identify male soccer players at risk of developing hamstring and quadriceps muscle injuries. PMID- 12531756 TI - Injury rates and profiles in female ice hockey players. AB - BACKGROUND: Little data exist on injury rates and profiles in female ice hockey players. OBJECTIVE: To examine the incidence of injury in female ice hockey players and compare injury rates with those of male players. STUDY DESIGN: Prospective cohort study. METHODS: Six male and six female teams from the Canada West Universities Athletic Association were followed prospectively for one varsity season. Preseason medical history forms were completed by each player. Injury report forms and attendance records for each team session were submitted by team therapists. RESULTS: Male players reported 161 injuries, whereas female players reported 66 injuries. However, the overall injury rates for male (9.19 injuries per 1000 athlete-exposures) and female (7.77 injuries per 1000 athlete exposures) players did not differ significantly. Ninety-six percent of injuries in female players and 79% in male players were related to contact mechanisms, even though intentional body checking is not allowed in female ice hockey. Women were more likely than men to be injured by contacting the boards or their opponent. Men sustained more severe injuries than women and missed about twice as many sessions (exposures) because of injury. Concussions were the most common injury in female players, followed by ankle sprains, adductor muscle strains, and sacroiliac dysfunction. CONCLUSION: Although the injury rate in female ice hockey players was expected to be lower than that in male players because of the lack of intentional body checking, the injury rates were found to be similar. PMID- 12531757 TI - The prevalence of glenohumeral osteoarthrosis in unstable shoulders. AB - BACKGROUND: Glenohumeral osteoarthritis has been a well-described complication of open procedures to correct shoulder instability. What remains unknown is whether chondral injuries sustained during instability episodes contribute to osteoarthritis or whether the cause is primarily the stabilization procedure itself. PURPOSE: To determine the prevalence of osteoarthrosis in a large database of patients with acute and chronic shoulder instability before primary stabilization surgery. STUDY DESIGN: Retrospective review of prospectively collected data. METHODS: Surgical and demographic data were collected on 422 patients with a diagnosis of shoulder instability who underwent arthroscopic examination and shoulder stabilization. RESULTS: There was a significant association between the grade of osteoarthrosis and the presence of osteoarthritis (grade III or IV chondral damage) with time from injury to surgery. Patients with osteoarthritis were significantly older than those without (34.9 versus 29.6 years). Multivariate analysis identified older age and time from injury to surgery as independent predictors of osteoarthritis. We found no association between direction of instability and the presence of osteoarthritis. CONCLUSION: The overall prevalence of glenohumeral osteoarthrosis was low. Predictors of osteoarthritis included time from injury to surgery and age. PMID- 12531758 TI - Glenohumeral translations are only partially restored after repair of a simulated type II superior labral lesion. AB - BACKGROUND: The effect on joint stability of repair of type II superior labrum and biceps anchor lesions is unknown. HYPOTHESIS: Increased translations of the glenohumeral joint after a simulated type II lesion will be reduced after the lesion is repaired. STUDY DESIGN: Controlled laboratory study. METHODS: A robotic/universal force-moment testing system was used to simulate load-and-shift and apprehension tests on eight cadaveric shoulders to determine joint kinematics of the shoulder after venting, creation of a type II lesion, and repair of the lesion. RESULTS: At 30 degrees of abduction, anterior translation of the vented joint in response to an anterior load was 18.7 +/- 8.5 mm and was significantly increased to 26.2 +/- 6.5 mm after simulation of a type II lesion. Repair did not restore anterior translation (23.9 +/- 8.6 mm) to that of the vented joint. The inferior translation that also occurred during application of an anterior load was 3.8 +/- 4.0 mm in the vented joint and increased significantly to 8.5 +/- 5.4 mm with a simulated type II lesion. After repair, the inferior translation decreased significantly to 6.7 +/- 5.3 mm. CONCLUSIONS: Repair of a type II lesion only partially restored glenohumeral translations to that of the vented joint. CLINICAL RELEVANCE: Surgical techniques including improved repair of passive stabilizers injured in the type II lesion should be considered. PMID- 12531759 TI - Capsular elongation in shoulders with recurrent anterior dislocation. Quantitative assessment with magnetic resonance arthrography. AB - BACKGROUND: Elongation of the shoulder capsule is often noticed on arthrograms or during surgery in shoulders of patients who have experienced recurrent anterior dislocations. HYPOTHESIS: We can quantify the elongation of the capsule in shoulders with recurrent anterior dislocations by using magnetic resonance arthrography. STUDY DESIGN: Retrospective review of prospectively collected data. METHODS: Twelve patients with unilateral recurrent anterior shoulder dislocations were enrolled in this study. Magnetic resonance images in the axial and coronal oblique planes were obtained from both shoulders (involved and uninvolved sides) after 10 ml of gadolinium/saline solution was injected into the glenohumeral joint. The length of the anteroinferior, inferior, and posteroinferior portions of the capsule was measured by using image analyzing software and normalized to the humeral head diameter. RESULTS: The anteroinferior capsule was significantly elongated in the involved shoulder at 4 mm (16% elongation) and 10 mm (19% elongation) superior to the inferior margin of the glenoid. The inferior capsule was also significantly elongated in the involved side both at the center (12% elongation) and at 4 mm anterior to the center of the glenoid (29% elongation). The posteroinferior capsule did not show any significant elongation. CONCLUSIONS: The anteroinferior and inferior portions of the shoulder capsule are elongated an average of 19% in shoulders with recurrent anterior dislocation. PMID- 12531760 TI - Comparison of clinical and dynamic knee function in patients with anterior cruciate ligament deficiency. AB - BACKGROUND: Whether passive measures of isokinetic muscle strength deficits and knee laxity are related to the dynamic function of the anterior cruciate ligament deficient knee remains unclear. HYPOTHESES: Arthrometer measurements are not predictive of peak external knee flexion moment (net quadriceps muscle moment), isokinetic quadriceps muscle strength correlates with peak external knee flexion moment (net quadriceps muscle moment), and isokinetic hamstring muscle strength correlates with peak external knee extension moment (net flexor muscle moment). STUDY DESIGN: Cross-sectional study. METHODS: Gait analysis was used to assess dynamic function during walking, jogging, and stair climbing in 44 subjects with unilateral anterior cruciate ligament deficiency and 44 control subjects. Passive knee laxity and isokinetic quadriceps and hamstring muscle strength were also measured. RESULTS: Arthrometer measurements did not correlate with peak external flexion or extension moments in any of the activities tested or with isokinetic quadriceps or hamstring muscle strength. Test subjects also had a significantly reduced peak external flexion moment during all three jogging activities and stair climbing compared with the control subjects and this was correlated with significantly reduced quadriceps muscle strength. CONCLUSIONS: Absolute knee laxity difference did not correlate with dynamic knee function as assessed by gait analysis and should not be used as a sole predictor for the outcome of treatment. Patients with greater than normal strength in the anterior cruciate ligament-deficient limb performed low- and high-stress activities in a more normal fashion than those with normal or less-than-normal strength. PMID- 12531761 TI - Three-dimensional tibiofemoral kinematics of the anterior cruciate ligament deficient and reconstructed knee during walking. AB - BACKGROUND: It is possible that gait abnormalities may play a role in the pathogenesis of meniscal or chondral injury as well as osteoarthritis of the knee in patients with anterior cruciate ligament deficiency. HYPOTHESIS: The three dimensional kinematics of anterior cruciate ligament-deficient knees are changed even during low-stress activities, such as walking, but can be restored by reconstruction. STUDY DESIGN: Case control study. METHODS: Using a three dimensional optoelectronic gait analysis system, we examined 13 patients with anterior cruciate ligament-deficient knees, 21 patients with anterior cruciate ligament-reconstructed knees, and 10 control subjects with uninjured knees during walking. RESULTS: Normal patterns of knee flexion-extension, abduction-adduction, and internal-external rotation during the gait cycle were maintained by all subjects. A significant difference in tibial rotation angle during the initial swing phase was found in anterior cruciate ligament-deficient knees compared with reconstructed and control knees. The patients with anterior cruciate ligament deficient knees rotated the tibia internally during the initial swing phase, whereas the others rotated externally. CONCLUSIONS: Patients with anterior cruciate ligament-deficient knees experienced repeated episodes of rotational instability during walking, whereas patients with reconstruction experienced tibial rotation that is closer to normal. CLINICAL RELEVANCE: Repeated episodes of knee rotational instability may play a role in the development of pathologic knee conditions. PMID- 12531762 TI - Effect of preexisting back pain on the incidence and severity of back pain in intercollegiate rowers. AB - BACKGROUND: Back pain is a common problem among rowers. PURPOSE: To determine whether preexisting back pain is a significant risk factor for back pain in intercollegiate rowers and how it relates to the ability of athletes to participate in college rowing. STUDY DESIGN: Survey. METHODS: Surveys from 1829 former intercollegiate rowing athletes were analyzed. The surveys included questions concerning back pain before the subjects' college career, back pain during intercollegiate rowing, missed practices, the duration of time lost from college rowing because of back pain, and career-ending back pain. Back pain was defined as pain lasting longer than 1 week. RESULTS: More subjects with preexisting back pain developed back pain during their college rowing career than subjects without preexisting back pain (57.1% versus 36.6%). Of those with preexisting pain, 55% missed practice because of back pain and 8% ended their college rowing careers; for those without preexisting pain, the percentages were 62% and 17%, respectively. For subjects with pain before their college rowing career, 78.8% missed less than 1 week and 5.9% missed more than 1 month. For subjects with no preexisting pain, 61.9% missed less than 1 week and 18.1% missed more than 1 month. CONCLUSIONS: While rowers with preexisting back pain are more likely to have back pain in college, they are less likely to miss extended periods of practice time or end their college rowing careers because of back pain. PMID- 12531763 TI - Reproducibility and reliability of the outerbridge classification for grading chondral lesions of the knee arthroscopically. AB - BACKGROUND: Few studies have investigated the accuracy and reproducibility of the Outerbridge classification system for classification of chondral damage in the knee. HYPOTHESIS: Arthroscopically assigned Outerbridge grades are accurate, reliable, and reproducible. STUDY DESIGN: Cadaver study. METHODS: Six cadaveric knees underwent diagnostic arthroscopy, which was videotaped. An arthrotomy was then performed and the arthroscopically identified lesions were measured with calipers. Nine orthopaedic surgeons reviewed each video and graded each chondral lesion two separate times. Accuracy of observations was calculated based on the percentage of agreement between the grades determined during arthroscopy and arthrotomy. RESULTS: The overall accuracy was 68% but varied by location. The kappa coefficient between the two scores was 0.602; the arthroscopy grade was higher than the arthrotomy grade 63% of the time. The intraobserver and interobserver kappa coefficients were 0.80 and 0.52, respectively. The mean interobserver kappa between the two physicians in practice 5 years or more was 0.72, compared with 0.50 for physicians in practice less than 5 years. CONCLUSIONS: The Outerbridge classification was moderately accurate when used to grade chondral lesions arthroscopically. CLINICAL RELEVANCE: Orthopaedic surgeons can accurately grade chondral lesions of the knee with the Outerbridge classification, regardless of their level of experience. PMID- 12531764 TI - Computer simulations of patellofemoral joint surgery. Patient-specific models for tuberosity transfer. AB - BACKGROUND: Variable clinical outcomes of tibial tuberosity transfer surgery have been reported. HYPOTHESES: The biomechanical outcome of surgery is patient specific; no single procedure produces superior results for all patients. Use of patient-specific computer models can optimize choice of procedure. STUDY DESIGN: Computer simulation study using clinical data. METHODS: We used patient-specific multibody models of the patellofemoral joints of 20 patients with a diagnosis of patellar subluxation and osteoarthritis. Four tibial tuberosity transfer procedures (two anterior and two anteromedial) were simulated for each patient and compared with their preoperative model. RESULTS: When results for all patients were averaged, all simulated operations produced a statistically significant decrease in surface-wide mean contact stress, although no significant difference was found among them. CONCLUSIONS: The simulated surgical outcomes were patient-specific: no single procedure was consistently superior at decreasing peak or mean stress and each procedure produced a potentially detrimental outcome, an increase in either mean stress or peak stress, in at least one patient. CLINICAL RELEVANCE: Computer simulation may serve as a valuable tool for tailoring procedures to specific patients. PMID- 12531765 TI - The effect of anterior cruciate ligament deficiency and functional bracing on translation of the tibia relative to the femur during nonweightbearing and weightbearing. AB - BACKGROUND: Much of what is known about functional bracing is derived from studies of the knee during unweighted or weightbearing conditions, whereas little is known about the transition between these postures. HYPOTHESIS: Bracing the anterior cruciate ligament-deficient knee during nonweightbearing, throughout the transition to weightbearing, and during weightbearing reduces the abnormal translations of the tibia relative to the femur to within the limits of normal. STUDY DESIGN: Controlled laboratory study. METHODS: Subjects with chronic anterior cruciate ligament tears were studied with and without braces. Anteroposterior shear and compressive loads were applied to the knee, and translations of the tibia relative to the femur were measured while subjects were nonweightbearing, throughout the transition to weightbearing, and during weightbearing. RESULTS: Bracing the anterior cruciate ligament-deficient knee resulted in a significant reduction of anteroposterior laxity values, to a level within the limits of the normal knee during nonweightbearing and weightbearing postures. In contrast, when the anterior cruciate ligament-deficient knees transitioned from nonweightbearing to weightbearing, the anterior translation of the tibia relative to the femur was 3.5 times greater than in the normal knee, and bracing did not reduce this to within the limits of normal. CONCLUSIONS: Bracing a knee with a chronic anterior cruciate ligament tear was effective in reducing abnormal anteroposterior laxity during nonweightbearing and weightbearing; however, braces were not effective in reducing the abnormal anterior translations produced by the change between these postures. CLINICAL RELEVANCE: This study explains why subjects with anterior cruciate ligament tears gain partial control of pathologic anteroposterior laxity with the use of a brace but may continue to experience abnormal translations during activity. PMID- 12531766 TI - Symptomatic, partial union of the hook of the hamate fracture in athletes. AB - BACKGROUND: Many clinicians believe that after fracture of the hook of the hamate, nonunions are the rule rather than the exception. PURPOSE: To describe a previously unreported condition in which the fractured hook of the hamate unites only along its ulnar column and causes pain similar to that in cases of nonunion. STUDY DESIGN: Retrospective review. METHODS: Eight patients who had sustained a hook of the hamate fracture were identified, on the basis of their persistent pain, as having achieved only partial union. This diagnosis was made despite evidence on computed tomography of healed bone across a significant portion of the fracture. All eight patients underwent subperiosteal excision of the hamulus because of persistent pain. At an average of 4 years after excision, each patient was evaluated by dynamometer grip strength testing and a subjective outcome assessment. RESULTS: All patients were able to return to their preinjury level of sports participation at an average 8 weeks after excision of the hook of the hamate. Grip strength was not adversely affected in any patient. Subjective evaluation of each patient revealed no residual pain. CONCLUSIONS: Chronic or unresolved pain on the ulnar side of the wrist after hook of the hamate fracture can be due to partial union. This condition should be managed no differently from a symptomatic nonunion, with excision of the hook of the hamate and repair of the ligament attachments. PMID- 12531767 TI - Quantitative assessment of classic anteroinferior bony Bankart lesions by radiography and computed tomography. AB - BACKGROUND: An anteroinferior osseous defect of the glenoid rim is sometimes encountered in patients with recurrent anterior dislocations of the shoulder. A defect of more than 21% of the glenoid length is reported to cause instability after Bankart repair. HYPOTHESIS: We can estimate the critical size of glenoid defects by using radiography or computed tomography. STUDY DESIGN: A controlled laboratory study. METHODS: Osseous defects of 0%, 9%, 21%, 34%, and 46% of the glenoid length were created stepwise in 12 cadaveric scapulae, and plain radiographs simulating the axillary and West Point views and computed tomographic images were obtained. The maximum width of the remnant glenoid was measured under each condition and expressed as a percentage of the width of the intact glenoid. RESULTS: A 21% defect appeared to be 18.6% of the intact glenoid on the West Point view. With computed tomography, a 21% defect resulted in loss of 50% of the width on a single slice across the lower one-fourth of the glenoid. CONCLUSIONS: We can estimate the size of a glenoid defect by using the West Point radiographic view or computed tomogram. CLINICAL RELEVANCE: These images gave decisive information as to whether an osseous glenoid defect required bone grafting to achieve stability after Bankart repair. PMID- 12531768 TI - Injuries among world-class professional beach volleyball players. The Federation Internationale de Volleyball beach volleyball injury study. AB - BACKGROUND: Very little is known about the injury characteristics of beach volleyball. PURPOSE: To describe the incidence and pattern of injuries among professional male and female beach volleyball players. STUDY DESIGN: Cohort study -retrospective injury recall and prospective registration. METHODS: Injuries occurring over a 7.5-week interval of the summer season were retrospectively registered by interviewing 178 of the 188 participating players (95%) in the 2001 Beach Volleyball World Championships. Injuries were also cataloged prospectively during five of the tournaments held during this interval. RESULTS: Fifty-four acute injuries was recorded, of which 23 (43%) resulted in 1 or more days of missed practice or competition. The incidence of acute time-loss injuries was estimated to be 3.1 per 1000 competition hours and 0.8 per 1000 training hours. Knee (30%), ankle (17%), and finger injuries (17%) accounted for more than half of all acute time-loss injuries. In addition, 67 players reported 79 overuse injuries for which they received medical attention during the study period. The three most common overuse conditions were low back pain (19%), knee pain (12%), and shoulder problems (10%). Similar results were observed in the prospective portion of the study. CONCLUSIONS: The rate of acute time-loss injuries in beach volleyball is considerably lower than that in most other team sports, but overuse injuries affecting the low back, knees, and shoulder represent a significant source of disability and impaired performance for professional beach volleyball players. PMID- 12531769 TI - Subscapularis muscle activity during selected rehabilitation exercises. AB - BACKGROUND: The upper and lower portions of the subscapularis muscle are independently innervated and activated. HYPOTHESIS: Upper and lower portions of the subscapularis muscle demonstrate different activation levels and require different exercises for rehabilitation. STUDY DESIGN: Controlled laboratory study. METHODS: Fifteen healthy subjects performed seven shoulder-strengthening exercises. Electromyographic data were collected from the latissimus dorsi, teres major, pectoralis major, infraspinatus, supraspinatus, and upper and lower subscapularis muscles. RESULTS: Upper subscapularis muscle activity was greater than lower subscapularis muscle activity for all exercises except for internal rotation with 0 degrees of humeral abduction. The push-up plus and diagonal exercises consistently stressed the upper and lower subscapularis muscles to the greatest extent. CONCLUSIONS: Humeral abduction was found to have a strong influence on the selective activation of the upper versus the lower subscapularis muscle and thus supported the design of different exercise continuums. In addition, the push-up plus and diagonal exercises were found to be superior to traditional internal rotation exercises for activating both functional portions of the subscapularis muscle. CLINICAL RELEVANCE: Our results showing that the upper and lower portions of the subscapularis muscle are functionally independent may affect training or rehabilitation protocols for the rotator cuff muscles. PMID- 12531770 TI - Posterior cruciate ligament injury and posterolateral instability in a 6-year-old child. A case report. PMID- 12531771 TI - Dissection of the proximal subclavian artery with consecutive thrombosis and embolic occlusion of the hand arteries after playing golf. PMID- 12531772 TI - Open repairs for the treatment of anterior shoulder instability. AB - Successful treatment of anterior instability of the shoulder requires a balance between restoring joint stability and minimizing loss of glenohumeral motion. The choice of treatment should be individualized on the basis of the patient's occupation and level of participation in sports, as well as on the degree of instability of the shoulder. Despite discussions to the contrary, there is no single "essential lesion," as proposed by Bankart, that is responsible for recurrent anterior shoulder instability, although the Bankart lesion is by far the most important. The choice of operative treatment must be tailored to correct the abnormality that is identified at the time of surgery. A variety of promising arthroscopic techniques have been developed for the treatment of anterior shoulder instability; however, open stabilization remains the standard, especially for severe instabilities, revision procedures, and for athletes who participate in contact sports. This article will review the open surgical techniques used for treatment of anterior instability of the shoulder. Both current and historical operations will be discussed. Regardless of which procedure is chosen by a surgeon, the treatment should follow the guidelines taught by Rowe: anatomic dissection at the time of surgery, identification and repair of the lesions responsible for the instability, returning tissues to their anatomic locations, and early postoperative range of motion. By following these guidelines, the results of treatment of anterior instability of the shoulder can be optimized. PMID- 12531773 TI - A review of the special tests associated with shoulder examination. Part I: the rotator cuff tests. AB - Careful examination of the shoulder is an essential component in forming a diagnosis of problems in this area. A number of tests have been described that are claimed to improve diagnostic accuracy by specifically examining one component of the shoulder complex. Many of these tests are eponymous, and there is confusion about not only how to perform them but also what conclusion to draw from the results. This article attempts to clarify the tests used to examine the rotator cuff by presenting them as described by the original authors with the additional aim of providing a source for those wishing to refresh their knowledge without the need to refer to the original source material. PMID- 12531774 TI - Interferon-gamma and interleukin-10 gene polymorphisms in pulmonary tuberculosis. AB - Several genes coding for different cytokines may affect host susceptibility to tuberculosis. This study investigates the relationship of the single base change polymorphic variants identified in the first intron of interferon-gamma (+874 T/A) and in the promoter region of interleukin-10 gene (-1,082 G/A), with cytokine production by peripheral blood mononuclear cells and tuberculosis susceptibility. We studied a Spanish population of 113 patients with culture proven pulmonary tuberculosis, 207 healthy close contacts (125 tuberculin reactive and 82 tuberculin negative), and 100 healthy tuberculin-negative control subjects. Multiple logistic regression analysis showed that individuals homozygous for the interferon-gamma (+874) A allele had a 3.75-fold increased risk of developing tuberculosis (95% confidence interval, 2.26-6.23, p = 0.0017). Stimulated production of interferon-gamma by peripheral mononuclear cells from patients with genotype AA was depressed compared with that of non-AA homozygotes at the time of diagnosis and after completion of therapy. Multivariate analysis showed that the presence of an AA genotype and the absolute number of lymphocytes were the only independent predictors of interferon-gamma production. In contrast, the different rates of interleukin-10 production associated with the interleukin 10 polymorphism did not affect susceptibility to tuberculosis. Thus, a genetic defect in the production of interferon-gamma in individuals homozygous for the (+874) A allele could contribute to their increased risk of developing tuberculosis. PMID- 12531775 TI - Effects of theophylline on ventilatory poststimulus potentiation in patients with brain damage. AB - Patients with brain damage, in contrast to normal subjects, exhibit a significant ventilatory undershoot when brief hypocapnic hypoxia is terminated abruptly by hyperoxia. This has been attributed to an impairment of activation of short-term potentiation, a brain stem mechanism promoting breathing stability. We hypothesized that in these patients theophylline, a drug that stabilizes breathing, may affect short-term potentiation. Eight stable patients with brain damage and 10 normal adults were studied. Activation of short-term potentiation was examined by brief exposure to hypoxia followed by hyperoxia after pretreatment with placebo or theophylline. Both in patients and normal subjects at the end of hypoxia ventilation increased to a similar magnitude with and without theophylline. In normal subjects independent of pretreatment, when hypoxia was terminated abruptly by hyperoxia, ventilation declined slowly to baseline without an undershoot, indicating activation of short-term potentiation. In patients with placebo, ventilation upon switching to hyperoxia exhibited a significant undershoot. This undershoot was significantly attenuated by theophylline, although compared with normal subjects, a slight hypoventilation was observed. We conclude that in patients with brain damage, theophylline largely prevents the hyperoxic drop of ventilation, presumably by affecting the activation of short-term potentiation. This may underlie the beneficial effect of theophylline on breathing stability. PMID- 12531777 TI - Airway structural alterations selectively associated with severe asthma. AB - To identify airway pathologic abnormalities selectively associated with severe asthma, we examined 10 control subjects, 10 patients with intermittent asthma, 15 patients with mild-to-moderate persistent asthma, 15 patients with severe persistent asthma, and 10 patients with chronic obstructive pulmonary disease. Bronchial biopsies were assessed for epithelial integrity; subepithelial basement membrane (SBM) thickness; collagen type III deposition; eosinophil, neutrophil, and fibroblast numbers; mucous gland and airway smooth muscle (ASM) areas; SBM ASM distance; ASM hypertrophy (increased cell size); and the expression of the contractile proteins alpha-actin, smooth muscle myosin heavy-chain isoforms, myosin light-chain kinase, and the phosphorylated form of the regulatory light chain of myosin. Neither mucosal eosinophilia nor neutrophilia, epithelial damage, or SBM thickness reflected asthma severity. In contrast, higher numbers of fibroblasts (p < 0.001), an increase in collagen type III deposition (p < 0.020), larger mucous gland (p < 0.040) and ASM (p < 0.001) areas, augmented ASM cell size (p < 0.001), and myosin light-chain kinase expression (p < 0.005) distinguished patients with severe persistent asthma from patients with milder disease or with chronic obstructive pulmonary disease. Stepwise multivariate regression analysis established that fibroblast numbers and ASM cell size were negatively associated with prebronchodilator and postbronchodilator FEV1 values in patients with asthma. We conclude that fibroblast accumulation and ASM hypertrophy in proximal airways are selective determinants of severe persistent asthma. PMID- 12531776 TI - Low isoniazid concentrations and outcome of tuberculosis treatment with once weekly isoniazid and rifapentine. AB - To understand why once-weekly isoniazid/rifapentine therapy for tuberculosis was less effective than twice-weekly isoniazid/rifampin, we studied human immunodeficiency virus-seronegative patients with either failure (n = 4), relapse (n = 35), or cure (n = 94), recruited from a comparative treatment trial. In multivariate analyses that were adjusted for severity of disease, low plasma concentrations of isoniazid were associated with failure/relapse with once-weekly isoniazid/rifapentine (median isoniazid area under the concentration-time curve for 12 hours after the dose [AUC(0-12)] was 36 microg x hour/ml in failure/relapse versus 56 microg x hour/ml in control cases p = 0.005), but not with twice-weekly isoniazid/rifampin. Furthermore, two patients who relapsed with Mycobacterium tuberculosis monoresistant to rifamycin had very low concentrations of isoniazid. Finally, isoniazid acetylator status determined by N acetyltransferase type 2 genotype was associated with outcome with once-weekly isoniazid/rifapentine (p = 0.03) but not twice-weekly isoniazid/rifampin. No rifamycin pharmacokinetic parameter was consistently and significantly associated with outcome (p > 0.10). Because low isoniazid concentrations were associated with failure/relapse, a drug with consistently greater area under the concentration-time curve than isoniazid may be needed to achieve highly active once-weekly therapy with rifapentine. PMID- 12531778 TI - Activation of dopamine D2-like receptors attenuates pulmonary C-fiber hypersensitivity in rats. AB - This study was performed to determine whether activation of dopamine D2-like receptors inhibits the hyperresponsiveness of pulmonary C fibers induced by inflammatory mediators such as prostaglandin E2 (PGE2). In anesthetized, open chest rats, constant infusion of PGE2 (1.5-4.5 microg/kg per minute, 2 minutes) significantly enhanced the C-fiber response to capsaicin injection. At 20 minutes after pretreatment with quinpirole (3 mg/kg, intravenous), a D2-like receptor agonist, the hyperresponsiveness to capsaicin of the same C fibers induced by PGE2 infusion was markedly attenuated, and this inhibitory effect lasted for more than 90 minutes. The effect of quinpirole was dose dependent and was antagonized by pretreatment with domperidone (5 mg/kg, intravenous), a D2-like receptor antagonist, administrated 10 minutes before the quinpirole injection. In a separate series of experiments, C-fiber responses to injections of phenyl biguanide and lactic acid and to constant-pressure lung inflation were augmented by PGE2; these potentiating responses were also significantly reduced by quinpirole. Furthermore, the effect of quinpirole was equally effective in inhibiting the increase in excitability of pulmonary C fibers induced by alveolar hypercapnia or constant infusion of adenosine. In conclusion, these results clearly show that activation of the dopamine D2-like receptors attenuates the hyperresponsiveness of pulmonary C fibers to both chemical stimuli and lung inflation. PMID- 12531779 TI - Hormone replacement therapy and sleep-disordered breathing. AB - Disordered breathing during sleep is more common among postmenopausal women than among their premenopausal counterparts, possibly because of declining levels of estrogen and progesterone. We examined the relationship between the use of replacement hormones and sleep-disordered breathing in a sample of 2,852 noninstitutionalized women, 50 years of age or older, who participated in the Sleep Heart Health Study. The frequency of apneas and hypopneas per hour of sleep (apnea-hypopnea index) was determined by unattended, single-night polysomnography at the participant's home. The prevalence of sleep-disordered breathing (apnea hypopnea index of 15 or more) among hormone users (61 of 907) was approximately half the prevalence among nonusers (286 of 1,945). Multivariable adjustment for known determinants of the disorder, including age, body mass index, and neck circumference, has attenuated the association, but only moderately (adjusted odds ratio, 0.55; 95% confidence interval, 0.41 to 0.75). The inverse association between hormone use and sleep-disordered breathing was evident in various subgroups and was particularly strong among women 50 to 59 years old (adjusted odds ratio, 0.36; 95% confidence interval, 0.21 to 0.60). If the observed associations are causal, hormone replacement therapy could have a role in preventing or alleviating sleep-disordered breathing. PMID- 12531780 TI - Effects of sex and of gene variants in constitutive nitric oxide synthases on exhaled nitric oxide. AB - Genetic factors may contribute to the variability of exhaled nitric oxide in healthy individuals. We studied exhaled nitric oxide and genetic variants in both neuronal and endothelial nitric oxide synthases in 105 healthy nonsmoking and smoking subjects. Genomic DNA was screened for a repeat polymorphism in intron 20 of the neuronal nitric oxide synthase gene and for the 894G/T mutation of the endothelial nitric oxide synthase gene. Exhaled nitric oxide was significantly higher in males than females among both nonsmokers (p < 0.0001) and smokers (p = 0.003). No association was found between exhaled nitric oxide and the endothelial nitric oxide synthase gene variant. However, healthy nonsmoking females with greater numbers of repeats (i.e., both alleles with 12 or more repeats) in neuronal nitric oxide synthase had significantly lower nitric oxide levels than did females with fewer numbers of repeats (i.e., at least one allele with fewer than 12 repeats) (13.6 +/- 1.6 versus 19.4 +/- 1.6 ppb, p = 0.02). No association was found between exhaled nitric oxide and neuronal nitric oxide synthase genotype in males. These data suggest that variants in the neuronal nitric oxide synthase gene contribute to the variability of airway nitric oxide concentrations in healthy females. PMID- 12531781 TI - Mechanism of acid adaptation of a fish living in a pH 3.5 lake. AB - Despite unfavorable conditions, a single species of fish, Osorezan dace, lives in an extremely acidic lake (pH 3.5) in Osorezan, Aomori, Japan. Physiological studies have established that this fish is able to prevent acidification of its plasma and loss of Na(+). Here we show that these abilities are mainly attributable to the chloride cells of the gill, which are arranged in a follicular structure and contain high concentrations of Na(+)-K(+)-ATPase, carbonic anhydrase II, type 3 Na(+)/H(+) exchanger (NHE3), type 1 Na(+)-HCO(3)(-) cotransporter, and aquaporin-3, all of which are upregulated on acidification. Immunohistochemistry established their chloride cell localization, with NHE3 at the apical surface and the others localized to the basolateral membrane. These results suggest a mechanism by which Osorezan dace adapts to its acidic environment. Most likely, NHE3 on the apical side excretes H(+) in exchange for Na(+), whereas the electrogenic type 1 Na(+)-HCO(3)(-) cotransporter in the basolateral membrane provides HCO(3)(-) for neutralization of plasma using the driving force generated by Na(+)-K(+)-ATPase and carbonic anhydrase II. Increased expression of glutamate dehydrogenase was also observed in various tissues of acid-adapted dace, suggesting a significant role of ammonia and bicarbonate generated by glutamine catabolism. PMID- 12531782 TI - Triggering of sarcoplasmic reticulum Ca2+ release and contraction by reverse mode Na+/Ca2+ exchange in trout atrial myocytes. AB - Whole cell patch clamp and intracellular Ca(2+) transients in trout atrial cardiomyocytes were used to quantify calcium release from the sarcoplasmic reticulum (SR) and examine its dependency on the Ca(2+) trigger source. Short depolarization pulses (2-20 ms) elicited large caffeine-sensitive tail currents. The Ca(2+) carried by the caffeine-sensitive tail current after a 2-ms depolarization was 0.56 amol Ca(2+)/pF, giving an SR Ca(2+) release rate of 279 amol Ca(2+). pF(-1). s(-1) or 4.3 mM/s. Depolarizing cells for 10 ms to different membrane potentials resulted in a local maximum of SR Ca(2+) release, intracellular Ca(2+) transient, and cell shortening at 10 mV. Although 100 microM CdCl(2) abolished this local maximum, it had no effect on SR Ca(2+) release elicited by a depolarization to 110 or 150 mV, and the SR Ca(2+) release was proportional to the membrane potential in the range -50 to 150 mV with 100 microM CdCl(2). Increasing the intracellular Na(+) concentration ([Na(+)]) from 10 to 16 mM enhanced SR Ca(2+) release but reduced cell shortening at all membrane potentials examined. In the absence of TTX, SR Ca(2+) release was potentiated with 16 mM but not 10 mM pipette [Na(+)]. Comparison of the total sarcolemmal Ca(2+) entry and the Ca(2+) released from the SR gave a gain factor of 18.6 +/- 7.7. Nifedipine (Nif) at 10 microM inhibited L-type Ca(2+) current (I(Ca)) and reduced the time integral of the tail current by 61%. The gain of the Nif sensitive SR Ca(2+) release was 16.0 +/- 4.7. A 2-ms depolarization still elicited a contraction in the presence of Nif that was abolished by addition of 10 mM NiCl(2). The gain of the Nif-insensitive but NiCl(2)-sensitive SR Ca(2+) release was 14.8 +/- 7.1. Thus both reverse-mode Na(+)/Ca(2+) exchange (NCX) and I(Ca) can elicit Ca(2+) release from the SR, but I(Ca) is more efficient than reverse-mode NCX in activating contraction. This difference may be due to extrusion of a larger fraction of the Ca(2+) released from the SR by reverse-mode NCX rather than a smaller gain for NCX-induced Ca(2+) release. PMID- 12531783 TI - Relative contribution of the TNF-alpha receptors to murine intimal hyperplasia. AB - Tumor necrosis factor-alpha (TNF-alpha) is an important mediator in the inflammatory response to vascular injury. The present study sought to determine the relative contribution of each TNF-alpha receptor subtype (p55 and p75) to intimal hyperplasia (IH) and characterize the mechanisms of transcriptional regulation after vascular injury. A murine model of wire carotid arterial injury was employed to induce IH in wild-type (WT), p55-deficient (p55-/-), and p75 deficient (p75-/-) mice. Compared with injured WT and p75-/- animals, p55-/- mice demonstrated a twofold reduction in IH. Additionally, p55-/- mice demonstrated a decrease in expression of nuclear factor-kappaB mRNA and protein. These observations suggest an important role for the p55 receptor in IH after mechanical endoluminal injury. Suppression of the transcriptional activator nuclear factor-kappaB may provide a mechanism by which p55-mediated IH is attenuated. PMID- 12531784 TI - Androgen-mediated induction of the kidney arachidonate hydroxylases is associated with the development of hypertension. AB - Hypertension is a leading cause of cardiovascular, cerebral, and renal disease morbidity and mortality, and epidemiological evidence suggests a role for sex dependent mechanisms in the pathophysiology of hypertension. We show here that treatment of rats with 5alpha-dihydrotestosterone increases the activity of the kidney arachidonate omega/omega-1 hydroxylase and the biosynthesis of 20-HETE (165 and 177% of control untreated male and female rats, respectively) and raises the systolic blood pressures of male and females rats by 46 and 57 mmHg, respectively. These androgen effects are associated with an upregulation in the kidney levels of CYP 4A8 mRNA and a decrease in CYP 4A1 transcripts. Dissected renal microvessels, the target tissue for most of the prohypertensive actions of 20-HETE, show an androgen-dependent upregulation of vascular CYP 4A8 mRNA and a fourfold increase in 20-HETE synthase activity. We propose that androgens regulate renal function and systemic blood pressure through a combination of transcriptional and hemodynamic mechanisms that are ultimately responsible for the regulation of renovascular tone and function. PMID- 12531785 TI - Angiotensin potentiates excitatory sensory synaptic transmission to medial solitary tract nucleus neurons. AB - Femtomole doses of angiotensin (ANG) II microinjected into nucleus tractus solitarii (nTS) decrease blood pressure and heart rate, mimicking activation of the baroreflex, whereas higher doses depress this reflex. ANG II might generate cardioinhibitory responses by augmenting cardiovascular afferent synaptic transmission onto nTS neurons. Intracellular recordings were obtained from 99 dorsal medial nTS region neurons in rat medulla horizontal slices to investigate whether ANG II modulated short-latency excitatory postsynaptic potentials (EPSPs) evoked by solitary tract (TS) stimulation. ANG II (200 fmol) increased TS-evoked EPSP amplitudes 20-200% with minimal membrane depolarization in 12 neurons excited by ANG II and glutamate, but not substance P (group A). Blockade of non-N methyl-d-aspartate receptors eliminated TS-evoked EPSPs and responses to ANG II. ANG II did not alter TS-evoked EPSPs in 14 other neurons depolarized substantially by ANG II and substance P (group B). ANG II appeared to selectively augment presynaptic sensory transmission in one class of nTS neurons but had only postsynaptic effects on another group of cells. Thus ANG II is likely to modulate cardiovascular function by more than one nTS neuronal pathway. PMID- 12531786 TI - Insulin-induced translocation of facilitative glucose transporters in fetal/neonatal rat skeletal muscle. AB - We examined the effect of insulin on fetal/neonatal rat skeletal muscle GLUT-1 and GLUT-4 concentrations and subcellular distribution by employing immunohistochemical analysis and subcellular fractionation followed by Western blot analysis. We observed that insulin did not alter total GLUT-1 or GLUT-4 concentrations or the GLUT-1 subcellular distribution in fetal/neonatal or adult skeletal muscle in 60 min. The basal and insulin-induced changes in subcellular distribution of GLUT-4 were different between the fetal/neonatal and adult skeletal muscle. Under basal conditions, sarcolemma-associated GLUT-4 was higher in the newborn compared with the adult, translating into a higher glucose transport. In contrast, insulin-induced translocation of GLUT-4 to the sarcolemma and insulin-induced glucose transport was lower in the newborn compared with the adult. This age-related change results in enhanced basal glucose transport to fuel myocytic proliferation and differentiation while relatively curbing the insulin-dependent glucose transport in the newborn. PMID- 12531787 TI - Neonatal or hepatocyte growth factor-potentiated adult gene therapy with a retroviral vector results in therapeutic levels of canine factor IX for hemophilia B. AB - Hemophilia B is a bleeding disorder resulting from factor IX (FIX) deficiency that might be treated with gene therapy. Neonatal delivery would correct the disease sooner than would transfer into adults, and could reduce immunological responses. Neonatal mice were injected intravenously with a Moloney murine leukemia virus-based retroviral vector (RV) expressing canine FIX (cFIX). They achieved 150% to 280% of normal cFIX antigen levels in plasma (100% is 5 microg/mL), which was functional in vitro and in vivo. Three newborn hemophilia B dogs that were injected intravenously with RV achieved 12% to 36% of normal cFIX antigen levels, which improved coagulation tests. Only one mild bleed has occurred during 14 total months of evaluation. This is the first demonstration of prolonged expression after neonatal gene therapy for hemophilia B in mice or dogs. Most animals failed to make antibodies to cFIX, demonstrating that neonatal gene transfer may induce tolerance. Although hepatocytes from newborns replicate, those from adults do not. Adult mice therefore received hepatocyte growth factor to induce hepatocyte replication prior to intravenous injection of RV. This resulted in expression of 35% of normal cFIX antigen levels for 11 months, although all mice produced anti-cFIX antibodies. This is the first demonstration that high levels of FIX activity can be achieved with an RV in adults without a partial hepatectomy to induce hepatocyte replication. We conclude that RV mediated hepatic gene therapy is effective for treating hemophilia B in mice and dogs, although the immune system may complicate gene transfer in adults. PMID- 12531788 TI - Expression of CD4 on human peripheral blood neutrophils. AB - CD4, the primary receptor for entry of HIV, is known to be expressed on T cells and monocytes/macrophages; healthy natural killer (NK) lymphocytes; in vitro human herpesvirus 6 (HHV6)-infected CD8+, NK, and gammadelta T lymphocytes; CD34+ progenitor cells; and a subset of eosinophils and basophils. We here report the unconventional expression of CD4 at the surface of peripheral blood neutrophils derived from 4 of 51 (7.8%) HIV-1-infected and 3 of 25 (12%) uninfected donors, with similar frequency within the 2 groups. The percentage of CD4+ neutrophils ranged from 39% to 97% of the total neutrophil population. Both surface and cytoplasmic forms of CD4 were present in neutrophils. Quantitative RNA polymerase chain reaction (PCR) revealed that neutrophils contain levels of CD4 mRNA comparable to those of peripheral blood mononuclear cells derived from the same donor. The conformation of CD4 expressed at the surface of neutrophils was similar to that of CD4 expressed on T lymphocytes as determined by the binding of monoclonal antibodies specific for conformational epitopes and the binding of recombinant HIV-1 gp120. Thus, our data provide evidence that neutrophils express endogenous CD4 and bind HIV. Owing to their abundance in peripheral blood, CD4+ neutrophils may influence significantly the biodistribution of HIV delivering it to sites of inflammation or to additional tissue reservoirs. PMID- 12531789 TI - Gene expression profile analysis of AIDS-related primary effusion lymphoma (PEL) suggests a plasmablastic derivation and identifies PEL-specific transcripts. AB - AIDS-related primary effusion lymphoma (PEL) is an HIV-associated malignancy characterized by the ability of the tumor cells to specifically home in the serous body cavities. Here we used gene expression profile analysis (about 12 000 genes) to further define the phenotype of PEL and to investigate the lymphoma relationship to normal B cells and to other tumor subtypes, including non-Hodgkin lymphomas (NHLs) of immunocompetent hosts and AIDS-associated NHL (AIDS-NHL). The results showed that PEL displayed a common gene expression profile that is clearly distinct from all NHLs of immunocompetent hosts and AIDS-NHL subtypes and, in contrast to those, is not related to germinal center (GC) or memory B cells. The gene expression profile of PEL was defined as plasmablastic because it showed features of both immunoblasts identified by Epstein-Barr virus (EBV) transformed lymphoblastoid cell lines and AIDS immunoblastic lymphoma, and plasma cells, as defined by multiple myeloma cell lines. Finally, our results identify a set of genes specifically expressed in PEL tumor cells. Their expression was validated at the protein level, suggesting their potential pathogenetic and clinical significance. PMID- 12531790 TI - Macrophage- and dendritic cell--dependent regulation of human B-cell proliferation requires the TNF family ligand BAFF. AB - Macrophages and dendritic cells play an important role in regulating B-cell responses, including proliferation to antigens such as trinitrophenyl (TNP) Ficoll and TNP-Brucella abortus. However, the mechanisms and molecule(s) that regulate these processes are relatively undefined. In this report, we show that human macrophages generated in vitro strongly costimulate proliferation of dense human tonsillar B cells ligated via their B-cell antigen receptor (BCR) but not proliferation via CD40. Similarly, dendritic cells also markedly enhance BCR activated B-cell proliferation. Soluble molecule(s) are required for human macrophages to costimulate proliferation of B cells triggered via their BCR. Importantly, a TACI (trans-membrane activator and CAML interactor)-Fc fusion protein inhibits both macrophage- and dendritic cell (DC)-dependent BCR-activated B-cell proliferation, indicating a requirement for at least one of the known TACI ligands, BAFF and/or APRIL. Consistent with a major role for BAFF, macrophages release BAFF at levels sufficient to potently costimulate BCR-induced B-cell proliferation. In addition, BAFF is more than 100-fold more potent than APRIL in enhancing BCR-mediated human B-cell proliferation. Furthermore, immunodepletion of APRIL under conditions that prevent APRIL-mediated B-cell costimulation does not block macrophage enhancement of B-cell proliferation. Finally, there is no correlation between the high levels of a proliferation-inducing ligand (APRIL) expressed by macrophages compared with DCs and the similar abilities of macrophages and DCs to enhance BCR-stimulated B-cell proliferation. In summary, our results suggest that macrophage- and DC-derived B-cell-activating factor belonging to the TNF family (BAFF) represents a key molecule by which macrophages and DCs directly regulate human B-cell proliferative responses to T-cell independent stimuli. PMID- 12531791 TI - Transfusion-transmitted cytomegalovirus infection after receipt of leukoreduced blood products. AB - Leukoreduced blood products are reportedly comparable to cytomegalovirus (CMV) seronegative products for the prevention of transfusion-transmitted CMV (TT-CMV) infection after stem cell (SC) transplantation. To determine if the incidence of TT-CMV was affected by the increasing use of leukoreduced blood products, we followed a prospective cohort of 807 CMV-seronegative SC transplant (SCT) recipients who underwent weekly surveillance using the pp65 antigenemia assay. The incidence of TT-CMV for 2 time periods was recorded: Period 1 (5/94-11/96), when only CMV-seronegative and/or filtered blood products were provided, and period 2 (12/96-2/00), when leukocyte-reduced platelets obtained by apheresis without filtration were also used. The incidence of TT-CMV was higher during period 2 (18/447, 4%) than period 1 (6/360, 1.7%) (P <.05); this was correlated with higher utilization of both filtered and apheresed products from CMV-positive donors in period 2. Multivariable analysis identified filtered red blood cell (RBC) units (but not apheresis platelet products) from CMV-positive donors as the primary predictor of TT-CMV: each additional filtered RBC unit was associated with a 32% increase in the odds for TT-CMV (95% confidence interval [CI]: 8%-61%, P =.006). Pre-emptive therapy with ganciclovir after detection of antigenemia prevented all but one case of CMV disease prior to day 100. CMV-seronegative products may thus be superior to leukoreduced products (particularly filtered RBCs) for the prevention of TT-CMV. In an era of "universal leukoreduction," the abandonment of CMV-seronegative inventories appears premature, particularly among populations at high risk of CMV disease that do not receive active surveillance. PMID- 12531792 TI - PDE4 inhibitors activate a mitochondrial apoptotic pathway in chronic lymphocytic leukemia cells that is regulated by protein phosphatase 2A. AB - Chronic lymphocytic leukemia (CLL) cells, but not peripheral blood T cells, undergo apoptosis following treatment with inhibitors of type 4 cyclic nucleotide phosphodiesterase (PDE4), a process that correlates dose dependently with elevation of adenosine 3',5'-cyclic monophosphate (cAMP) in leukemic cells. We show that treatment of CLL cells with rolipram, a prototypic PDE4 inhibitor, and forskolin, an adenylate cyclase activator, induces mitochondrial depolarization, release of cytochrome c into the cytosol, caspase-9 and -3 activation, and cleavage of poly(adenosine diphosphate [ADP]-ribose)polymerase. Inhibitors of caspase-9, but not caspase-8, block rolipram/forskolin-induced CLL apoptosis. In a subset of CLL patients, B-cell lymphoma 2 (Bcl-2)-associated death promoter homolog (Bad), a proapoptotic Bcl-2 family member that when phosphorylated on specific serine residues is sequestered in the cytosol by 14-3-3, was dephosphorylated at Ser112 following rolipram/forskolin treatment of leukemic cells. Rolipram/forskolin treatment also induced Bad to accumulate in CLL heavy membrane fractions, consistent with Bad translocation to mitochondria. To determine the mechanism for rolipram/forskolin-induced Bad dephosphorylation, we examined CLL phosphatase activity. Rolipram/forskolin treatment augmented protein phosphatase 2A (PP2A) activity, as well as levels of immunoreactive PP2A catalytic subunit. Treatment of CLL cells with a concentration of okadaic acid (5 nM) that selectively inhibits PP2A, reduced both rolipram/forskolin-induced mitochondrial cytochrome c release and mitochondrial depolarization. Okadaic acid restored Bad Ser112 phosphorylation and Bad association with 14-3-3 in rolipram/forskolin-treated CLL cells. These results suggest that PDE4 inhibitors may induce CLL apoptosis by activating PP2A-induced dephosphorylation of proapoptotic BH3-only Bcl-2 family members such as Bad. PMID- 12531793 TI - Arsenic trioxide-induced apoptosis in myeloma cells: p53-dependent G1 or G2/M cell cycle arrest, activation of caspase-8 or caspase-9, and synergy with APO2/TRAIL. AB - Arsenic trioxide (ATO) has been shown to induce differentiation and apoptosis in acute promyelocytic leukemia (APL) cells concomitant with down-regulation of the PML-RARalpha fusion protein, a product of the t(15:17) translocation characteristic of APL leukemic cells. However, ATO is also a potent inducer of apoptosis in a number of other cancer cells lacking the t(15:17) translocation. The exact mechanism of ATO-induced apoptosis in these cells is not yet clear. We tested the effect of ATO on 7 myeloma cell lines with varying p53 status and report that in cells with mutated p53, ATO induced rapid and extensive (more than 90%) apoptosis in a time- and dose-dependent manner concomitant with arrest of cells in G(2)/M phase of the cell cycle. Myeloma cells with wild-type (wt) p53 were relatively resistant to ATO with maximal apoptosis of about 40% concomitant with partial arrest of cells in G(1) and up-regulation of p21. The use of caspase blocking peptides, fluorescence-tagged caspase-specific substrate peptides, and Western immunoblotting confirmed the involvement of primarily caspase-8 and -3 in ATO-induced apoptosis in myeloma cells with mutated p53 and primarily caspase-9 and -3 in cells expressing wt p53. We also observed up-regulation by ATO of R1 and R2 APO2/TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) receptors. Most important, however, we observed a synergy between ATO and APO2/TRAIL in the induction of apoptosis in the partially resistant myeloma cell lines and in myeloma cells freshly isolated from myeloma patients. Our results justify the use of the combination of these 2 drugs in clinical setting in myeloma patients. PMID- 12531795 TI - Targeting of the collagen-binding site on glycoprotein VI is not essential for in vivo depletion of the receptor. AB - Glycoprotein (GP) VI is an essential collagen receptor on platelets and may serve as an attractive target for antithrombotic therapy. We have previously shown that a monoclonal antibody (mAb) against the major collagen-binding site on mouse GPVI (JAQ1) induces irreversible down-regulation of the receptor and, consequently, long-term antithrombotic protection in vivo. To determine whether this unique in vivo effect of JAQ1 is based on its interaction with the ligand-binding site on GPVI, we generated new mAbs against different epitopes on GPVI (JAQ2, JAQ3) and tested their in vitro and in vivo activity. We show that none of the mAbs inhibited platelet activation by collagen or the collagen-related peptide in vitro. Unexpectedly, however, injection of either antibody induced depletion of GPVI with the same efficacy and kinetics as JAQ1. Importantly, this effect was also seen with monovalent F(ab) fragments of JAQ2 and JAQ3, excluding the involvement of the Fc part or the dimeric form of anti-GPVI antibodies in this process. This indicates that anti-GPVI agents, irrespective of their binding site may generally induce down-regulation of the receptor in vivo. PMID- 12531794 TI - CHOP is superior to CNOP in elderly patients with aggressive lymphoma while outcome is unaffected by filgrastim treatment: results of a Nordic Lymphoma Group randomized trial. AB - This study was designed to test the hypothesis that administration of granulocyte colony-stimulating factor (G-CSF; filgrastim) during induction chemotherapy with CHOP (cyclophosphamide, vincristine, doxorubicin, prednisone) or CNOP (doxorubicin replaced with mitoxantrone) in elderly patients with aggressive non Hodgkin lymphoma (NHL) improves time to treatment failure (TTF), complete remission (CR) rate, and overall survival (OS). Furthermore, the efficacy of CHOP versus CNOP chemotherapy was compared. A total of 455 previously untreated patients older than 60 years with stages II to IV aggressive NHL were included in the analysis. Patients (median age, 71 years; range, 60-86 years) were randomized to receive CHOP (doxorubicin 50 mg/m(2)) or CNOP (mitoxantrone 10 mg/m(2)) with or without G-CSF (5 microg/kg from day 2 until day 10-14 of each cycle every 3 weeks; 8 cycles). Forty-seven patients previously hospitalized for class I to II congestive heart failure were randomized to receive CNOP with or without G-CSF (not included in the CHOP versus CNOP analysis). The CR rates in the CHOP/CNOP plus G-CSF and CHOP/CNOP groups were the same, 52%, and in the CHOP with or without G-CSF and CNOP with or without G-CSF groups, 60% and 43% (P <.001), respectively. No benefit of G-CSF in terms of TTF and OS could be shown (P =.96 and P =.22, respectively), whereas CHOP was superior to CNOP (TTF/OS P <.001). The incidences of severe granulocytopenia (World Health Organization grade IV) and granulocytopenic infections were higher in patients not receiving G-CSF. The cumulative proportion of patients receiving 90% or more of allocated chemotherapy was higher (P <.05) in patients receiving G-CSF. Concomitant G-CSF treatment did not improve CR rate, TTF, or OS. Patients receiving CHOP fared better than those given CNOP chemotherapy. The addition of G-CSF reduces the incidence of severe granulocytopenia and infections in elderly patients with aggressive NHL receiving CHOP or CNOP chemotherapy. PMID- 12531796 TI - The heat-shock protein receptor CD91 is up-regulated in monocytes of HIV-1 infected "true" long-term nonprogressors. AB - A small proportion of patients with human immunodeficiency virus type 1 (HIV-1) remains asymptomatic for a long period after infection. It is thought that a vigorous immune response may contribute to long-term nonprogression, though studies are confounded by heterogeneity among patients. We studied the levels of HIV-1 receptors, costimulatory T-cell molecules, and dendritic cell (DC) numbers in 18 patients with long-term infection, CD4 counts greater than 400 cells/mm(3), and HIV-1 viral loads lower than 50 copies/mL. These patients were further differentiated through the presence or absence of 2-LTR DNA circles, a possible marker for residual ongoing HIV-1 replication. A statistically significant increase in levels of CD91, the heat-shock protein (HSP) receptor, was observed in therapy-naive patients who had no evidence of ongoing viral replication (P =.01). This difference was most notable on their monocytes. High levels of CD91 may be a host factor that contributes to the maintenance of long-term nonprogression. The ability of CD91 to internalize alpha-defensins and to cross present exogenous antigen to cytotoxic T lymphocytes through major histocompatibility complex (MHC) class 1 may maintain CD8(+) responses in these patients. PMID- 12531797 TI - Beta 2-microglobulin as a negative regulator of the immune system: high concentrations of the protein inhibit in vitro generation of functional dendritic cells. AB - Two common features in human immunodeficiency virus infection and acquired immunodeficiency syndrome, rheumatoid arthritis, and hematologic malignancies including multiple myeloma are elevated serum levels of beta(2)-microglobulin (beta(2)M) and activation or inhibition of the immune system. We hypothesized that beta(2)M at high concentrations may have a negative impact on the immune system. In this study, we examined the effects of beta(2)M on monocyte-derived dendritic cells (MoDCs). The addition of beta(2)M (more than 10 microg/mL) to the cultures reduced cell yield, inhibited the up-regulation of surface expression of human histocompatibility leukocyte antigen (HLA)-ABC, CD1a, and CD80, diminished their ability to activate T cells, and compromised generation of the type-1 T cell response induced in allogeneic mixed-lymphocyte reaction. Compared with control MoDCs, beta(2)M-treated cells produced more interleukin-6 (IL-6), IL-8, and IL-10. beta(2)M-treated cells expressed significantly fewer surface CD83, HLA ABC, costimulatory molecules, and adhesion molecules and were less potent at stimulating allospecific T cells after an additional 48-hour culture in the presence of tumor necrosis factor-alpha and IL-1beta. During cell culture, beta(2)M down-regulated the expression of phosphorylated mitogen-activated protein (MAP) kinases, extracellular signal-related kinase (ERK), and mitogen induced extracellular kinase (MEK), inhibited nuclear factor-kappaB (NF-kappaB), and activated signal transducer and activator of transcription-3 (STAT3) in treated cells, all of which are involved in cell differentiation and proliferation. Thus, our study demonstrates that beta(2)M at high concentrations retards the generation of MoDCs, which may involve down-regulation of major histocompatibility complex class I molecules, inactivation of Raf/MEK/ERK cascade and NF-kappaB, and activation of STAT3, and it merits further study to elucidate the underlying mechanisms. PMID- 12531798 TI - Rapamycin inhibits IL-4--induced dendritic cell maturation in vitro and dendritic cell mobilization and function in vivo. AB - Rapamycin (RAPA) is a potent immunosuppressive macrolide hitherto believed to mediate its action primarily via suppression of lymphocyte responses to interleukin 2 (IL-2) and other growth factors. We show here that this view is incomplete and provide evidence that RAPA suppresses the functional activation of dendritic cells (DCs) both in vitro and in vivo. In vitro, RAPA inhibits IL-4 dependent maturation and T-cell stimulatory activity of murine bone marrow derived DCs. These effects are associated with posttranscriptional down regulation of both subunits of the IL-4 receptor complex (CD124, CD132) and are mediated via binding of RAPA to its intracellular receptor FK506-binding protein 12 (FKBP12). In vivo, RAPA impairs steady-state DC generation and fms-like tyrosine 3 kinase ligand (Flt3L)-induced DC mobilization. In addition, in vivo administration of RAPA impairs DC costimulatory molecule up-regulation, production of proinflammatory cytokines, and T-cell allostimulatory capacity. These novel findings have implications for RAPA-based therapy of chronic DC triggered autoimmune diseases, transplant rejection, and hematologic malignancies with activating Flt3 mutations. PMID- 12531799 TI - Molecular sequelae of histone deacetylase inhibition in human malignant B cells. AB - Histone acetylation modulates gene expression, cellular differentiation, and survival and is regulated by the opposing activities of histone acetyltransferases (HATs) and histone deacetylases (HDACs). HDAC inhibition results in accumulation of acetylated nucleosomal histones and induces differentiation and/or apoptosis in transformed cells. In this study, we characterized the effect of suberoylanilide hydroxamic acid (SAHA), the prototype of a series of hydroxamic acid-based HDAC inhibitors, in cell lines and patient cells from B-cell malignancies, including multiple myeloma (MM) and related disorders. SAHA induced apoptosis in all tumor cells tested, with increased p21 and p53 protein levels and dephosphorylation of Rb. We also detected cleavage of Bid, suggesting a role for Bcl-2 family members in regulation of SAHA-induced cell death. Transfection of Bcl-2 cDNA into MM.1S cells completely abrogated SAHA induced apoptosis, confirming its protective role. SAHA did not induce cleavage of caspase-8, -9, or -3 in MM.1S cells during the early phase of apoptosis, and the pan-caspase inhibitor ZVAD-FMK did not protect against SAHA. Conversely, poly(ADP)ribose polymerase (PARP) was cleaved in a pattern indicative of calpain activation, and the calpain inhibitor calpeptin abrogated SAHA-induced cell death. Importantly, SAHA sensitized MM.1S cells to death receptor-mediated apoptosis and inhibited the secretion of interleukin 6 (IL-6) induced in bone marrow stromal cells (BMSCs) by binding of MM cells, suggesting that it can overcome cell adhesion-mediated drug resistance. Our studies delineate the mechanisms whereby HDAC inhibitors mediate anti-MM activity and overcome drug resistance in the BM milieu and provide the framework for clinical evaluation of SAHA, which is bioavailable, well tolerated, and bioactive after oral administration, to improve patient outcome. PMID- 12531800 TI - Rituximab for the treatment of refractory autoimmune hemolytic anemia in children. AB - Autoimmune hemolytic anemia (AIHA) in children is sometimes characterized by a severe course, requiring prolonged administration of immunosuppressive therapy. Rituximab is able to cause selective in vivo destruction of B lymphocytes, with abrogation of antibody production. In a prospective study, we have evaluated the use of rituximab for the treatment of AIHA resistant to conventional treatment. Fifteen children with AIHA were given rituximab, 375 mg/m(2)/dose for a median of 3 weekly doses. All patients had previously received 2 or more courses of immunosuppressive therapy; 2 patients had undergone splenectomy. After completing treatment, all children received intravenous immunoglobulin for 6 months. Treatment was well tolerated. With a median follow-up of 13 months, 13 patients (87%) responded, whereas 2 patients did not show any improvement. Median hemoglobin levels increased from 7.7 g/dL to a 2-month posttreatment level of 11.8 g/dL (P <.001). Median absolute reticulocyte counts decreased from 236 to 109 x 10(9)/L (P <.01). An increase in platelet count was observed in patients with concomitant thrombocytopenia (Evans syndrome). Three responder patients had relapse, 7, 8, and 10 months after rituximab infusion, respectively. All 3 children received a second course of rituximab, again achieving disease remission. Our data indicate that rituximab is both safe and effective in reducing or even abolishing hemolysis in children with AIHA and that a sustained response can be achieved in the majority of cases. Disease may recur, but a second treatment course may be successful in controlling the disease. PMID- 12531802 TI - Retraction. Neutrophil elastase mutations in severe congenital neutropenia patients of the original Kostmann family. PMID- 12531801 TI - Continuous absence of metaphase-defined cytogenetic abnormalities, especially of chromosome 13 and hypodiploidy, ensures long-term survival in multiple myeloma treated with Total Therapy I: interpretation in the context of global gene expression. AB - Metaphase cytogenetic abnormalities (CAs), especially of chromosome 13 (CA 13), confer a grave prognosis in multiple myeloma even with tandem autotransplantations as applied in Total Therapy I, which enrolled 231 patients between 1989 and 1994. With a median follow-up of almost 9 years, the prognostic implications of all individual CAs, detected prior to treatment and at relapse, were investigated. Among all CAs and standard prognostic factors examined prior to therapy, only hypodiploidy and CA 13 (hypo-13 CA), alone or in combination, were associated with shortest event-free survival and overall survival (OS). The shortest postrelapse OS was observed with hypo-13 CA, which was newly detected in 18 of all 28 patients presenting with this abnormality at relapse. Superior prognosis was associated with the absence of any CA at both diagnosis and relapse (10-year OS, 40%). The lack of independent prognostic implications of other CAs points to a uniquely aggressive behavior of hypo-13 CA (present in 16% of patients at diagnosis). With the use of microarray data in 146 patients enrolled in Total Therapy II, overexpression of cell cycle genes distinguished CA from no CA, especially in cases of del(13) detected by interphase fluorescence in situ hybridization (FISH). FISH 13, resulting in a haploinsufficiency of RB1 and other genes mapping to chromosome 13, as well as activation of IGF1R, appears to have an amplifying effect on cell cycle gene expression, thus providing a molecular explanation for the dire outcome of patients with CA 13 compared with those with other CAs. PMID- 12531803 TI - Responding naive T cells differ in their sensitivity to Fas engagement: early death of many T cells is compensated by costimulation of surviving T cells. AB - Engagement of Fas (CD95) induces death of activated T cells but can also potentiate T-cell response to CD3 ligation. Yet, the effects of Fas-mediated signals on activation of naive T cells have remained controversial. We followed naive T cells responding under Fas ligation. Ligation of Fas simultaneously with activation by antigen-bearing dendritic cells promoted early death in half of the responding naive murine CD4 T cells. Surprisingly, it simultaneously accelerated cell division and interferon-gamma (IFN-gamma) production among surviving T cells. These cells developed quickly an activation-associated phenotype (CD44(hi), CD62L(lo)), responded vigorously to antigen rechallenge, were partially resistant to subsequent induction of cell death via Fas, and were long lived in vivo. Compared with cells becoming apoptotic, the surviving cells expressed lower levels of Fas and higher levels of T-cell receptor (TCR), CD4, and interleukin-2 receptor (IL-2R). Their survival was associated with expression of antiapoptotic cellular FLICE-inhibitory protein (c-FLIP), Bcl-X(L), and Bcl-2. Thus, at the time of T-cell activation there is a subtle balance in the effects of Fas ligation that differs on a cell-to-cell basis. Factors that predict cell survival include expression levels of Fas, TCR, CD4, and IL-2R. Early death of some cells and a pronounced response of the surviving cells suggest that Fas ligation can both up- and down-regulate a primary T-cell response. PMID- 12531804 TI - Kaposi sarcoma-associated viral cyclin K overrides cell growth inhibition mediated by oncostatin M through STAT3 inhibition. AB - DNA viruses have evolved a number of mechanisms to inhibit the major cellular tumor-suppressor pathways. Viral oncogenes can override growth suppressive signals and extend the virus proliferative capacity. The Kaposi sarcoma associated human herpesvirus 8 (KSHV) encodes a protein, cyclin K, that is similar to cellular cyclin D1 but behaves atypically. Cyclin K resists the actions of the p16 INK4a and p27Kip1 inhibitors and extends the range of cdk6 substrates, thereby inducing cell-cycle progression toward S phase. In this study, we show that cyclin K overrides growth suppressive signals through signal transducer and activator of transcription 3 (STAT3) inactivation. Cyclin K was found to associate with the activation domain of STAT3 to inhibit its DNA-binding and transcriptional activities. Overexpression of cyclin K and inhibition of STAT3 prevents the growth suppressive effect imposed by the interleukin 6-type cytokine, oncostatin M. Altogether, these results suggest that KSHV is able to override growth suppressive effects through multiple mechanisms, and they further indicate that cyclin K plays an important role in the oncogenic activity of these viruses. PMID- 12531805 TI - SU11248 is a novel FLT3 tyrosine kinase inhibitor with potent activity in vitro and in vivo. AB - FLT3 (fms-related tyrosine kinase/Flk2/Stk-2) is a receptor tyrosine kinase (RTK) primarily expressed on hematopoietic cells. In blasts from acute myelogenous leukemia (AML) patients, 2 classes of FLT3 activating mutations have been identified: internal tandem duplication (ITD) mutations in the juxtamembrane domain (25%-30% of patients) and point mutations in the kinase domain activation loop (7%-8% of patients). FLT3-ITD mutations are the most common molecular defect identified in AML and have been shown to be an independent prognostic factor for decreased survival. FLT3-ITD is therefore an attractive molecular target for therapy. SU11248 is a recently described selective inhibitor with selectivity for split kinase domain RTKs, including platelet-derived growth factor receptors, vascular endothelial growth factor receptors, and KIT. We show that SU11248 also has potent activity against wild-type FLT3 (FLT3-WT), FLT3-ITD, and FLT3 activation loop (FLT3-Asp835) mutants in phosphorylation assays. SU11248 inhibits FLT3-driven phosphorylation and induces apoptosis in vitro. In addition, SU11248 inhibits FLT3-induced VEGF production. The in vivo efficacy of SU11248 was investigated in 2 FLT3-ITD models: a subcutaneous tumor xenograft model and a bone marrow engraftment model. We show that SU11248 (20 mg/kg/d) dramatically regresses FLT3-ITD tumors in the subcutaneous tumor xenograft model and prolongs survival in the bone marrow engraftment model. Pharmacokinetic and pharmacodynamic analysis in subcutaneous tumors showed that a single administration of an efficacious drug dose potently inhibits FLT3-ITD phosphorylation for up to 16 hours following a single dose. These results suggest that further exploration of SU11248 activity in AML patients is warranted. PMID- 12531806 TI - G-protein-coupled receptor signaling in Syk-deficient neutrophils and mast cells. AB - The Syk tyrosine kinase is essential for immunoreceptor and multiple integrin functions as well as being implicated in signaling from G-protein-coupled receptors (GPCR) in cell lines, transfection systems, and pharmacologic studies. In contrast, using Syk-deficient primary cells, we show here that Syk does not play a major functional role in chemoattractant/chemokine signaling in neutrophils and mast cells. syk(-/-) neutrophils showed normal respiratory burst and degranulation in response to the bacterial peptide formyl-Met-Leu-Phe (fMLP). The migration of neutrophils toward fMLP was similarly not affected by the syk(-/ ) mutation. fMLP initiated normal Ca(2+)-signal, activation of the extracellular signal-related kinase (ERK) and p38 mitogen-activated protein (MAP) kinase cascades, and polymerization of cellular actin in the absence of Syk. syk(-/-) and wild-type neutrophils also responded similarly to LTB(4), C5a, and the chemokines macrophage inflammatory protein-1 (MIP-1)alpha or MIP-2, both in functional assays and in intracellular signaling mechanisms. Furthermore, bone marrow-derived syk(-/-) mast cells showed normal activation of the Akt, ERK, and p38 MAP kinase pathways when stimulated by the GPCR ligand adenosine. We conclude that, in contrast to previous reports, Syk does not play a major role in GPCR signaling. PMID- 12531807 TI - Role of the MyD88 transduction signaling pathway in endothelial activation by antiphospholipid antibodies. AB - Antiphospholipid syndrome (APS) is an autoimmune disease characterized by the persistent presence of antiphospholipid antibodies (aPLs) and recurrent thrombosis or fetal loss. The thrombophilic state has been partially related to the induction of a proinflammatory and procoagulant endothelial cell (EC) phenotype induced by anti-beta(2)-glycoprotein I (beta(2)-GPI) antibodies that bind beta(2)-GPI expressed on the EC surface. Anti-beta(2)-GPI antibody binding has been shown to induce nuclear factor-kappa B (NF-kappa B) translocation leading to a proinflammatory EC phenotype similar to that elicited by interaction with microbial products (lipopolysaccharide [LPS]) and proinflammatory cytokines (interleukin 1 beta [IL-1 beta], tumor necrosis factor alpha [TNF-alpha]). However, the upstream signaling events are not characterized yet. To investigate the endothelial signaling cascade activated by anti-beta(2)-GPI antibodies, we transiently cotransfected immortalized human microvascular endothelial cells (HMEC-1) with dominant-negative constructs of different components of the pathway (Delta TRAF2, Delta TRAF6, Delta MyD88) together with reporter genes (NF-kappa B luciferase and pCMV-beta-galactosidase). Results showed that both human anti beta(2)-GPI IgM monoclonal antibodies as well as polyclonal affinity-purified anti-beta(2)-GPI IgG display a signaling cascade comparable to that activated by LPS or IL-1. Delta TRAF6 and Delta MyD88 significantly abrogate antibody-induced as well as IL-1- or LPS-induced NF-kappa B activation, whereas Delta TRAF2 (involved in NF-kappa B activation by TNF) does not affect it. Moreover, anti- beta(2)-GPI antibodies and LPS followed the same time kinetic of IL-1 receptor activated kinase (IRAK) phosphorylation, suggesting an involvement of the toll like receptor (TLR) family. Our findings demonstrate that anti-beta(2)-GPI antibodies react with their antigen likely associated to a member of the TLR/IL-1 receptor family on the EC surface and directly induce activation. PMID- 12531808 TI - Granulocyte colony-stimulating factor and the risk of secondary myeloid malignancy after etoposide treatment. AB - Event-free survival for children with acute lymphoblastic leukemia (ALL) now exceeds 80% in the most effective trials. Failures are due to relapse, toxicity, and second cancers such as therapy-related myeloid leukemia or myelodysplasia (t ML). Topoisomerase II inhibitors and alkylators can induce t-ML; additional risk factors for t-ML remain poorly defined. The occurrence of t-ML among children who had received granulocyte colony-stimulating factor (G-CSF) following ALL remission induction therapy prompted us to examine this and other putative risk factors for t-ML in 412 children treated on 2 consecutive ALL protocols from 1991 to 1998. All children received etoposide and anthracyclines, 99 of whom received G-CSF; 284 also received cyclophosphamide, 58 of whom also received cranial irradiation. There were 20 children who developed t-ML at a median of 2.3 years (range, 1.0-6.0 years), including 16 cases of acute myeloid leukemia, 3 myelodysplasia, and 1 chronic myeloid leukemia. Stratifying by protocol, the cumulative incidence functions differed (P =.017) according to the use of G-CSF and irradiation: 6-year cumulative incidence (standard error) of t-ML of 12.3% (5.3%) among the 44 children who received irradiation without G-CSF, 11.0% (3.5%) among the 85 children who received G-CSF but no irradiation, 7.1% (7.2%) among the 14 children who received irradiation plus G-CSF, and 2.7% (1.3%) among the 269 children who received neither irradiation nor G-CSF. Even when children receiving irradiation were excluded, the incidence was still higher in those receiving G-CSF (P =.019). In the setting of intensive antileukemic therapy, short-term use of G-CSF may increase the risk of t-ML. PMID- 12531809 TI - Dexamethasone versus prednisone and daily oral versus weekly intravenous mercaptopurine for patients with standard-risk acute lymphoblastic leukemia: a report from the Children's Cancer Group. AB - Conventional therapy for childhood acute lymphoblastic leukemia (ALL) includes prednisone and oral 6-mercaptopurine. Prior observations suggested potential advantages for dexamethasone over prednisone and for intravenous (IV) over oral 6 mercaptopurine, which remain to be validated. We report the results of a randomized trial of more than 1000 subjects that examined the efficacy of dexamethasone and IV 6-mercaptopurine. Children with National Cancer Institute standard-risk ALL were randomly assigned in a 2 x 2 factorial design to receive dexamethasone (6 mg/m(2)/d) for 28 days in induction, plus taper, compared with prednisone (40 mg/m(2)/d). The second randomized assignment was for daily oral or weekly IV 6-mercaptopurine during consolidation. During maintenance, 5 days of the randomized steroid was given monthly, at the same dose, and all patients received daily oral 6-mercaptopurine. During delayed intensification, all patients received a dexamethasone dosage of 10 mg/m(2)/d for 21 days, with taper. Intrathecal (IT) methotrexate was the sole central nervous system-directed therapy. Patients randomly assigned to receive dexamethasone had a 6-year isolated central nervous system-relapse rate of 3.7% +/- 0.8%, compared with 7.1% +/- 1.1% for prednisone (P =.01). There was also a trend toward fewer isolated bone marrow relapses with dexamethasone. The 6-year event-free survival (EFS) was 85% +/- 2% for dexamethasone and 77% +/- 2% for prednisone (P =.002). EFS was similar with oral or IV 6-mercaptopurine; however, patients assigned to IV 6 mercaptopurine had decreased survival after relapse. PMID- 12531810 TI - Free radical stress in chronic lymphocytic leukemia cells and its role in cellular sensitivity to ROS-generating anticancer agents. AB - 2-Methoxyestradiol (2-ME), a new anticancer agent currently in clinical trials, has been demonstrated to inhibit superoxide dismutase (SOD) and to induce apoptosis in leukemia cells through a free radical-mediated mechanism. Because the accumulation of superoxide (O(2)-) by inhibition of SOD depends on the cellular generation of O(2)-, we hypothesized that the endogenous production of superoxide may be a critical factor that affects the antileukemia activity of 2 ME. In the present study, we investigated the relationship between cellular O(2)- contents and the cytotoxic activity of 2-ME in primary leukemia cells from 50 patients with chronic lymphocytic leukemia (CLL). Quantitation of O(2)- revealed that the basal cellular O(2)- contents are heterogeneous among patients with CLL. The O(2)- levels were significantly higher in CLL cells from patients with prior chemotherapy. CLL cells with higher basal O(2)- contents were more sensitive to 2 ME in vitro than those with lower O(2)- contents. There was a significant correlation between the 2-ME-induced O(2)- increase and the loss of cell viability. Importantly, addition of arsenic trioxide, a compound capable of causing reactive oxygen species (ROS) generation, significantly enhanced the activity of 2-ME, even in the CLL cells that were resistant to 2-ME alone. These results suggest that the cellular generation of O(2)- plays an important role in the cytotoxic action of 2-ME and that it is possible to use exogenous ROS producing agents such as arsenic trioxide in combination with 2-ME to enhance the antileukemia activity and to overcome drug resistance. Such a combination strategy may have potential clinical applications. PMID- 12531812 TI - Utility of FDG-PET scanning in lymphoma by WHO classification. AB - We retrospectively evaluated (18)fluoro-2-deoxyglucose positron emission tomography (FDG-PET) scans in 172 patients with lymphoma and correlated results with pathologic diagnosis using the World Health Organization (WHO) classification system. In total, FDG-PET detected disease in at least one site in 161 patients (94%) and failed to detect disease in 11 patients (6%). The most frequent lymphoma diagnoses were diffuse large B-cell lymphoma (LBCL; n = 51), Hodgkin lymphoma (HL; n = 47), follicular lymphoma (FL; n = 42), marginal zone lymphoma (MZL; n = 12), mantle cell lymphoma (MCL; n = 7), and peripheral T-cell lymphoma (PTCL; n = 5). FDG-PET detected disease in 100% of patients with LBCL and MCL and in 98% of patients with HL and FL. In contrast, FDG-PET detected disease in only 67% of MZL and 40% of PTCL. Comparison with bone marrow biopsies showed that FDG-PET was not reliable for detection of bone marrow involvement in any lymphoma subtype. PMID- 12531811 TI - Excess hemoglobin digestion and the osmotic stability of Plasmodium falciparum infected red blood cells. AB - During their asexual reproduction cycle (about 48 hours) in human red cells, Plasmodium falciparum parasites consume most of the host cell hemoglobin, far more than they require for protein biosynthesis. They also induce a large increase in the permeability of the host cell plasma membrane to allow for an increased traffic of nutrients and waste products. Why do the parasites digest hemoglobin in such excess? And how can infected red cells retain their integrity for the duration of the asexual cycle when comparably permeabilized uninfected cells hemolyse earlier? To address these questions we encoded the multiplicity of factors known to influence host cell volume in a mathematical model of the homeostasis of a parasitized red cell. The predicted volume changes were subjected to thorough experimental tests by monitoring the stage-related changes in the osmotic fragility of infected red cell populations. The results supported the model predictions of biphasic volume changes comprising transient shrinkage of infected cells with young trophozoites followed by continuous volume increase to about 10% lower than the critical hemolytic volume of approximately 150 fL by the end of the asexual cycle. Analysis of these results and of additional model predictions demonstrated that the osmotic stability of infected red cells can be preserved only by a large reduction in impermeant solute concentration within the host cell compartment. Thus, excess hemoglobin consumption represents an essential evolutionary strategy to prevent the premature hemolysis of the highly permeabilized infected red cell. PMID- 12531813 TI - Late-onset X-linked sideroblastic anemia following hemodialysis. AB - X-linked sideroblastic anemia (XLSA) is due to deficient activity of erythroid specific 5-aminolevulinate synthase (ALAS2). We report here a patient who developed sideroblastic anemia at the age of 81 years while undergoing hemodialysis. The diagnosis of sideroblastic anemia was established by the presence of ringed sideroblasts in the bone marrow, and treatment with oral pyridoxine completely eliminated the ringed sideroblasts. We identified a novel point mutation in the fifth exon of this patient's ALAS2 gene, which resulted in an amino acid change at residue 159 from aspartic acid to asparagine (Asp159Asn). In vitro analyses of recombinant Asp159Asn ALAS2 revealed that this mutation accounted for the pyridoxine-responsiveness of this disease. The very late onset in this case of XLSA emphasizes that nutritional deficiencies caused either by dietary irregularities in the elderly or, as in this case, by maintenance hemodialysis therapy, may uncover occult inherited enzymatic deficiencies in the heme biosynthetic pathway. PMID- 12531815 TI - Patterns of somatic mutations in VH genes reveal pathways of clonal transformation from MGUS to multiple myeloma. AB - Monoclonal gammopathy of undetermined significance (MGUS) can transform to multiple myeloma (MM). In myeloma, mutated V(H) genes with sequence homogeneity reveal a postfollicular origin. Previously, some MGUS cases showed mutated V(H) genes with intraclonal variation, indicating an earlier stage of arrest. We investigated progression from 2 of 2 MGUS to MM, in which V(H) genes confirmed clonal evolution. In one MGUS case, intraclonal heterogeneity was evident, and transformation to myeloma occurred rapidly with apparent homogeneity in the emergent clone. However, residual MGUS-derived sequences were detectable at this time. Heterogeneity in MGUS does not associate with benign disease, but it indicates an origin from a tumorigenic cell, most likely surface immunoglobulin(+), undergoing somatic mutation. The remaining case displayed intraclonal homogeneity at the MGUS stage, conceivably resulting from a self cloning outgrowth from MGUS with heterogeneity. Transformation can occur at either MGUS stage, but it involves a single cell in which somatic mutation is then silent. PMID- 12531814 TI - A band 3-based macrocomplex of integral and peripheral proteins in the RBC membrane. AB - We have studied the membrane proteins of band 3 anion exchanger (AE1)-deficient mouse and human red blood cells. It has been shown previously that proteins of the band 3 complex are reduced or absent in these cells. In this study we show that proteins of the Rh complex are also greatly reduced (Rh-associated glycoprotein, Rh polypeptides, CD47, glycophorin B) or absent (LW). These observations suggest that the Rh complex is associated with the band 3 complex in healthy RBCs. Mouse band 3(-/-) RBCs differed from the human band 3-deficient RBCs in that they retained CD47. Aquaporin 1 was reduced, and its glycosylation was altered in mouse and human band 3-deficient RBCs. Proteins of the glycophorin C complex, and other proteins with independent cytoskeletal interactions, were present in normal or increased amounts. To obtain direct evidence for the association of the band 3 and the Rh protein complexes in the RBC, we examined whether Rh complex proteins were coimmunoprecipitated with band 3 from membranes. RhAG and Rh were found to be efficiently coimmunoprecipitated with band 3 from deoxycholate-solubilized membranes. Results suggest that band 3 forms the core of a macrocomplex of integral and peripheral RBC membrane proteins. The presence of these proteins in a single structural macrocomplex makes it likely that they have linked functional or regulatory roles. We speculate that this macrocomplex may function as an integrated CO(2)/O(2) gas exchange unit (metabolon) in the erythrocyte. PMID- 12531817 TI - Israel considers paying people for donating a kidney. PMID- 12531816 TI - ICAM-directed vascular immunotargeting of antithrombotic agents to the endothelial luminal surface. AB - Drug targeting to a highly expressed, noninternalizable determinant up-regulated on the perturbed endothelium may help to manage inflammation and thrombosis. We tested whether inter-cellular adhesion molecule-1 (ICAM-1) targeting is suitable to deliver antithrombotic drugs to the pulmonary vascular lumen. ICAM-1 antibodies bind to the surface of endothelial cells in culture, in perfused lungs, and in vivo. Proinflammatory cytokines enhance anti-ICAM binding to the endothelium without inducing internalization. (125)I-labeled anti-ICAM and a reporter enzyme (beta-Gal) conjugated to anti-ICAM bind to endothelium and accumulate in the lungs after intravenous administration in rats and mice. Anti ICAM is seen to localize predominantly on the luminal surface of the pulmonary endothelium by electron microscopy. We studied the pharmacological effect of ICAM directed targeting of tissue-type plasminogen activator (tPA). Anti-ICAM/tPA, but not control IgG/tPA, conjugate accumulates in the rat lungs, where it exerts plasminogen activator activity and dissolves fibrin microemboli. Therefore, ICAM may serve as a target for drug delivery to endothelium, for example, for pulmonary thromboprophylaxis. Enhanced drug delivery to sites of inflammation and the potential anti-inflammatory effect of blocking ICAM-1 may enhance the benefit of this targeting strategy. PMID- 12531818 TI - Monitoring the marketing of infant formula feeds. PMID- 12531819 TI - Preventing skin cancer. PMID- 12531820 TI - Levelling the playing field for regulation of nicotine. PMID- 12531821 TI - Doctors and managers. PMID- 12531824 TI - Medicines Control Agency must be more open. PMID- 12531822 TI - Preventing and treating tetanus. PMID- 12531826 TI - Company launches campaign to "counter" BMJ claims. PMID- 12531827 TI - Environmentalist accused of scientific dishonesty. PMID- 12531828 TI - Government publishes strategy to improve diabetes care. PMID- 12531829 TI - Nigerian immunisation programme sees results. PMID- 12531838 TI - UK doctors warned after ricin poison found in police raid. PMID- 12531839 TI - FDA insists that oestrogen products for menopause carry a warning. PMID- 12531840 TI - Painting is earliest example of portrayal of Down s syndrome. PMID- 12531842 TI - Monitoring compliance with the International Code of Marketing of Breastmilk Substitutes in west Africa: multisite cross sectional survey in Togo and Burkina Faso. AB - OBJECTIVES: To monitor compliance with the International Code of Marketing of Breastmilk Substitutes in health systems, sales outlets, distribution points, and the news media in Togo and Burkina Faso, west Africa. DESIGN: Multisite cross sectional survey. PARTICIPANTS: Staff at 43 health facilities and 66 sales outlets and distribution points, 186 health providers, and 105 mothers of infants aged < or =5 months in 16 cities. RESULTS: Six (14%) health facilities had received donations of breast milk substitutes. All donations were being given to mothers free of charge. Health providers in five (12%) health facilities had received free samples of breast milk substitutes for purposes other than professional research or evaluation. Health professionals in five (12%) health facilities had received promotional gifts from manufacturers. Promotional materials of commercial breast milk substitutes were found in seven (16%) health facilities. Special displays to market commercial breast milk substitutes were found in 29 (44%) sales and distribution points. Forty commercial breast milk substitutes violated the labelling standards of the code: 21 were manufactured by Danone, 11 by Nestle, and eight by other national and international manufacturers. Most (148, 90%) health providers had never heard of the code, and 66 mothers (63%) had never received any counselling on breast feeding by their health providers. CONCLUSION: In west Africa manufacturers are violating the code of marketing of breast milk substitutes. Comparable levels of code violations are observed with (Burkina Faso) or without (Togo) regulating legislation. Legislation must be accompanied by effective information, training, and monitoring systems to ensure that healthcare providers and manufacturers comply with evidence based practice and the code. PMID- 12531843 TI - Folate and vitamin B-12 and risk of fatal cardiovascular disease: cohort study from Busselton, Western Australia. AB - OBJECTIVE: To test the hypothesis that the incidence of fatal coronary heart disease and cardiovascular disease in a general population is related to serum and red cell folate and vitamin B-12 concentrations. DESIGN: Cohort study with follow up of 29 years. SETTING: Busselton, Western Australia. PARTICIPANTS: 1419 men and 1531 women aged 20 to 90 years, who were alive more than three years after their participation in the 1969 Busselton health survey. 2314 (78.4%) had no cardiovascular disease at the initial survey. MAIN OUTCOME MEASURES: Hazard ratios for fatal coronary heart disease and cardiovascular disease in men and women according to baseline concentrations of serum and red cell folate and serum vitamin B-12. RESULTS: 213 men and 159 women died from coronary heart disease, and 342 men and 302 women died from cardiovascular disease. Serum and red cell folate concentrations showed a moderate positive correlation (r=0.26, P<0.001) but otherwise serum and red cell folate and serum B-12 concentrations were not strongly correlated with each other or with other standard risk factors. After age and standard risk factors were adjusted for, there was no independent association between folate and B-12 concentrations and death from coronary heart disease or cardiovascular disease in the full cohort or the subcohort with no cardiovascular disease at baseline. The multivariate adjusted hazard ratio for death from cardiovascular disease in the lowest versus the highest category of red cell folate concentration was 1.05 (95% confidence interval 0.77 to 1.43) in men and 1.10 (0.81 to 1.51) in women. CONCLUSIONS: These findings do not support the hypothesis that lower folate and B-12 concentrations increase the risk of fatal cardiovascular disease in a general population. The routine use of these vitamins for preventing cardiovascular disease should await evidence from clinical trials. PMID- 12531844 TI - Impact of changing diagnostic criteria on incidence, management, and outcome of acute myocardial infarction: retrospective cohort study. PMID- 12531845 TI - Demographic and Health Surveys: caesarean section rates in sub-Saharan Africa. PMID- 12531846 TI - Effect of fetal sex on labour and delivery: retrospective review. PMID- 12531847 TI - Why do general practitioners prescribe antibiotics for sore throat? Grounded theory interview study. AB - OBJECTIVES: To understand why general practitioners prescribe antibiotics for some cases of sore throat and to explore the factors that influence their prescribing. DESIGN: Grounded theory interview study. SETTING: General practice. PARTICIPANTS: 40 general practitioners: 25 in the maximum variety sample and 15 in the theoretical sample. RESULTS: General practitioners are uncertain which patients will benefit from antibiotics but prescribe for sicker patients and for patients from socioeconomically deprived backgrounds because of concerns about complications. They are also more likely to prescribe in pressured clinical contexts. Doctors are mostly comfortable with their prescribing decisions and are not prescribing to maintain the doctor-patient relationship. CONCLUSIONS: General practitioners have reduced prescribing for sore throat in response to research and policy initiatives. Further interventions to reduce prescribing would need to improve identification of patients at risk of complications and be workable in busy clinical situations. PMID- 12531848 TI - Transferability of principles of evidence based medicine to improve educational quality: systematic review and case study of an online course in primary health care. PMID- 12531849 TI - Management of the severely malnourished child: perspective from developing countries. PMID- 12531850 TI - Unsuspected haemophilia in children with a single swollen joint. PMID- 12531851 TI - Anticoagulation in hospitals and general practice. PMID- 12531853 TI - Distinguishing between salt poisoning and hypernatraemic dehydration in children. PMID- 12531854 TI - Mobile phone use has not replaced smoking in adolescence. PMID- 12531855 TI - Medical school applications. Pool of potential medical students needs to be enlarged. PMID- 12531856 TI - Public access defibrillators. Potential efficacy of public access defibrillation may be underestimated. PMID- 12531857 TI - Society of Homeopaths does not advise against vaccination. PMID- 12531858 TI - Consuming alcohol on duty may have adverse consequences. PMID- 12531859 TI - Risk assessment scales poorly predict pressure ulceration. PMID- 12531860 TI - Papua New Guinea needs law and order above all. PMID- 12531861 TI - Free smoke alarms: a fire officer responds. PMID- 12531862 TI - Mastectomy is not always so bad. PMID- 12531863 TI - Theme issue for medics and health informed public. Maybe not such a good idea. PMID- 12531864 TI - Emotional intelligence. PMID- 12531866 TI - Rare genetic mutations shed light on the pathogenesis of Parkinson disease. PMID- 12531867 TI - Programmed cell death in amyotrophic lateral sclerosis. PMID- 12531868 TI - Oxidative stress and nitration in neurodegeneration: cause, effect, or association? PMID- 12531869 TI - A role for nitric oxide in hydroxyurea-mediated fetal hemoglobin induction. PMID- 12531870 TI - The irresistible biology of resistin. PMID- 12531871 TI - Genotype, phenotype: upstairs, downstairs in the family of cardiomyopathies. PMID- 12531872 TI - Enumerating autoreactive T cells in peripheral blood: a big step in diabetes prediction. PMID- 12531873 TI - The intricate interplay among body weight, stress, and the immune response to friend or foe. PMID- 12531874 TI - Disruption of the CXCR4/CXCL12 chemotactic interaction during hematopoietic stem cell mobilization induced by GCSF or cyclophosphamide. AB - Hematopoietic progenitor cells (HPCs) normally reside in the bone marrow (BM) but can be mobilized into the peripheral blood (PB) after treatment with GCSF or chemotherapy. In previous studies, we showed that granulocyte precursors accumulate in the BM during mobilization induced by either GCSF or cyclophosphamide (CY), leading to the accumulation of active neutrophil proteases in this tissue. We now report that mobilization of HPCs by GCSF coincides in vivo with the cleavage of the N-terminus of the chemokine receptor CXCR4 on HPCs resident in the BM and mobilized into the PB. This cleavage of CXCR4 on mobilized HPCs results in the loss of chemotaxis in response to the CXCR4 ligand, the chemokine stromal cell-derived factor-1 (SDF-1/CXCL12). Furthermore, the concentration of SDF-1 decreased in vivo in the BM of mobilized mice, and this decrease coincided with the accumulation of serine proteases able to directly cleave and inactivate SDF-1. Since both SDF-1 and its receptor, CXCR4, are essential for the homing and retention of HPCs in the BM, the proteolytic degradation of SDF-1, together with that of CXCR4, could represent a critical step leading to the mobilization of HPCs into the PB in response to GCSF or CY. PMID- 12531875 TI - Defective TNF-alpha-mediated hepatocellular apoptosis and liver damage in acidic sphingomyelinase knockout mice. AB - This study addressed the contribution of acidic sphingomyelinase (ASMase) in TNF alpha-mediated hepatocellular apoptosis. Cultured hepatocytes depleted of mitochondrial glutathione (mGSH) became sensitive to TNF-alpha, undergoing a time dependent apoptotic cell death preceded by mitochondrial membrane depolarization, cytochrome c release, and caspase activation. Cyclosporin A treatment rescued mGSH-depleted hepatocytes from TNF-alpha-induced cell death. In contrast, mGSH depleted hepatocytes deficient in ASMase were resistant to TNF-alpha-mediated cell death but sensitive to exogenous ASMase. Furthermore, although in vivo administration of TNF-alpha or LPS to galactosamine-pretreated ASMase(+/+) mice caused liver damage, ASMase(-/-) mice exhibited minimal hepatocellular injury. To analyze the requirement of ASMase, we assessed the effect of glucosylceramide synthetase inhibition on TNF-alpha-mediated apoptosis. This approach, which blunted glycosphingolipid generation by TNF-alpha, protected mGSH-depleted ASMase(+/+) hepatocytes from TNF-alpha despite enhancement of TNF-alpha stimulated ceramide formation. To further test the involvement of glycosphingolipids, we focused on ganglioside GD3 (GD3) because of its emerging role in apoptosis through interaction with mitochondria. Analysis of the cellular redistribution of GD3 by laser scanning confocal microscopy revealed the targeting of GD3 to mitochondria in ASMase(+/+) but not in ASMase(-/-) hepatocytes. However, treatment of ASMase(-/-) hepatocytes with exogenous ASMase induced the colocalization of GD3 and mitochondria. Thus, ASMase contributes to TNF-alpha-induced hepatocellular apoptosis by promoting the mitochondrial targeting of glycosphingolipids. PMID- 12531876 TI - Idiopathic restrictive cardiomyopathy is part of the clinical expression of cardiac troponin I mutations. AB - Restrictive cardiomyopathy (RCM) is an uncommon heart muscle disorder characterized by impaired filling of the ventricles with reduced volume in the presence of normal or near normal wall thickness and systolic function. The disease may be associated with systemic disease but is most often idiopathic. We recognized a large family in which individuals were affected by either idiopathic RCM or hypertrophic cardiomyopathy (HCM). Linkage analysis to selected sarcomeric contractile protein genes identified cardiac troponin I (TNNI3) as the likely disease gene. Subsequent mutation analysis revealed a novel missense mutation, which cosegregated with the disease in the family (lod score: 4.8). To determine if idiopathic RCM is part of the clinical expression of TNNI3 mutations, genetic investigations of the gene were performed in an additional nine unrelated RCM patients with restrictive filling patterns, bi-atrial dilatation, normal systolic function, and normal wall thickness. TNNI3 mutations were identified in six of these nine RCM patients. Two of the mutations identified in young individuals were de novo mutations. All mutations appeared in conserved and functionally important domains of the gene. This article was published online in advance of the print edition. The date of publication is available from the JCI website, http://www.jci.org. PMID- 12531877 TI - Prediction of spontaneous autoimmune diabetes in NOD mice by quantification of autoreactive T cells in peripheral blood. AB - Autoimmune (type 1) diabetes mellitus results from the destruction of insulin producing pancreatic beta cells by T lymphocytes. Prediction of cell-mediated autoimmune diseases by direct detection of autoreactive T cells in peripheral blood has proved elusive, in part because of their low frequency and reduced avidity for peptide MHC ligands. This article was published online in advance of the print edition. The date of publication is available from the JCI website, http://www.jci.org. PMID- 12531878 TI - Adipose-derived resistin and gut-derived resistin-like molecule-beta selectively impair insulin action on glucose production. AB - The adipose-derived hormone resistin is postulated to link obesity to insulin resistance and diabetes. Here, the infusion of either resistin or the resistin like molecule-beta (RELMbeta) rapidly induced severe hepatic but not peripheral insulin resistance. In the presence of physiologic hyperinsulinemia, the infusion of purified recombinant resistin, increasing circulating resistin levels by approximately twofold to 15-fold, inhibited glucose metabolism such that lower rates of glucose infusion were required to maintain the plasma glucose concentration at basal levels. The effects of resistin and RELMbeta on in vivo insulin action were completely accounted for by a marked increase in the rate of glucose production. These results support the notion that a novel family of fat- and gut-derived circulating proteins modulates hepatic insulin action. PMID- 12531880 TI - Leptin surge precedes onset of autoimmune encephalomyelitis and correlates with development of pathogenic T cell responses. AB - In the work presented here, we explored the influence of leptin on the kinetics of experimental autoimmune encephalomyelitis (EAE) onset, in the EAE-associated inflammatory anorexia, and in the development of pathogenic T cell responses. We found that the expression of serum leptin increased before the clinical onset of EAE in disease-susceptible C57BL/6J (H-2(b)) and SJL/J (H-2(s)) strains of mice, which are models of chronic-progressive and relapsing-remitting EAE, respectively. This increase in serum leptin correlated with disease susceptibility, reduction in food intake, and decrease in body weight. Indeed, acute starvation, which is able to prevent the increase in serum leptin, delayed disease onset and attenuated clinical symptoms by inducing a T helper 2 cytokine switch. Furthermore, immunohistochemical analysis revealed a parallel in situ production of leptin in inflammatory infiltrates and in neurons only during the acute/active phase of both chronic-progressive and relapsing-remitting EAE. We also found that leptin secretion by activated T cells sustained their proliferation in an autocrine loop, since antileptin receptor antibodies were able to inhibit the proliferative response of autoreactive T cells in vitro. Given that leptin appears to regulate EAE susceptibility, inflammatory anorexia, and pathogenic T-cell immune function, we postulate that it may offer a potential target in the treatment of multiple sclerosis. PMID- 12531879 TI - Hydroxyurea induces fetal hemoglobin by the nitric oxide-dependent activation of soluble guanylyl cyclase. AB - Hydroxyurea treatment of patients with sickle-cell disease increases fetal hemoglobin (HbF), which reduces hemoglobin S polymerization and clinical complications. Despite its use in the treatment of myeloproliferative diseases for over 30 years, its mechanism of action remains uncertain. Recent studies have demonstrated that hydroxyurea generates the nitric oxide (NO) radical in vivo, and we therefore hypothesized that NO-donor properties might determine the hemoglobin phenotype. We treated both K562 erythroleukemic cells and human erythroid progenitor cells with S-nitrosocysteine (CysNO), an NO donor, and found similar dose- and time-dependent induction of gamma-globin mRNA and HbF protein as we observed with hydroxyurea. Both hydroxyurea and CysNO increased cGMP levels, and the guanylyl cyclase inhibitors ODQ, NS 2028, and LY 83,538 abolished both the hydroxyurea- and CysNO-induced gamma-globin expression. These data provide strong evidence for an NO-derived mechanism for HbF induction by hydroxyurea and suggest possibilities for therapies based on NO-releasing or potentiating agents. PMID- 12531881 TI - Injection of genetically engineered fibroblasts corrects regenerated human epidermolysis bullosa skin tissue. AB - Current therapeutic strategies for genetic skin disorders rely on the complex process of grafting genetically engineered tissue to recipient wound beds. Because fibroblasts synthesize and secrete extracellular matrix, we explored their utility in recessive dystrophic epidermolysis bullosa (RDEB), a blistering disease due to defective extracellular type VII collagen. Intradermal injection of RDEB fibroblasts overexpressing type VII collagen into intact RDEB skin stably restored correctly localized type VII collagen expression in vivo and normalized hallmark RDEB disease features, including subepidermal blistering and anchoring fibril defects. This article was published online in advance of the print edition. The date of publication is available from the JCI website, http://www.jci.org. PMID- 12531882 TI - Burst-like control of lipolysis by the sympathetic nervous system in vivo. AB - Rapid oscillations of visceral lipolysis have been reported. To examine the putative role of the CNS in oscillatory lipolysis, we tested the effects of beta(3)-blockade on pulsatile release of FFAs. Arterial blood samples were drawn at 1-minute intervals for 120 minutes from fasted, conscious dogs (n = 7) during the infusion of saline or bupranolol (1.5 micro g/kg/min), a high-affinity beta(3)-blocker. FFA and glycerol time series were analyzed and deconvolution analysis was applied to estimate the rate of FFA release. During saline infusion FFAs and glycerol oscillated in phase at about eight pulses/hour. Deconvolution analysis showed bursts of lipolysis (nine pulses/hour) with time-dependent variation in burst frequency. Bupranolol completely removed rapid FFA and glycerol oscillations. Despite removal of lipolytic bursts, plasma FFAs (0.31 mM) and glycerol (0.06 mM) were not totally suppressed and deconvolution analysis revealed persistent non-oscillatory lipolysis (0.064 mM/min). These results show that lipolysis in the fasting state consists of an oscillatory component, which appears to be entirely dependent upon sympathetic innervation of the adipose tissue, and a non-oscillatory, constitutive component, which persists despite beta(3)-blockade. The extinction of lipid fuel bursts by beta(3)-blockade implies a role for the CNS in the maintenance of cyclic provision of lipid fuels. PMID- 12531883 TI - A methylated oligonucleotide inhibits IGF2 expression and enhances survival in a model of hepatocellular carcinoma. AB - IGF-II is a mitogenic peptide that has been implicated in hepatocellular oncogenesis. Since the silencing of gene expression is frequently associated with cytosine methylation at cytosine-guanine (CpG) dinucleotides, we designed a methylated oligonucleotide (MON1) complementary to a region encompassing IGF2 promoter P4 in an attempt to induce DNA methylation at that locus and diminish IGF2 mRNA levels. MON1 specifically inhibited IGF2 mRNA accumulation in vitro, whereas an oligonucleotide (ON1) with the same sequence but with nonmethylated cytosines had no effect on IGF2 mRNA abundance. MON1 treatment led to the specific induction of de novo DNA methylation in the region of IGF2 promoter hP4. Cells from a human hepatocellular carcinoma (HCC) cell line, Hep 3B, were implanted into the livers of nude mice, resulting in the growth of large tumors. Animals treated with MON1 had markedly prolonged survival as compared with those animals treated with saline or a truncated methylated oligonucleotide that did not alter IGF2 mRNA levels in vitro. This study demonstrates that a methylated sense oligonucleotide can be used to induce epigenetic changes in the IGF2 gene and that inhibition of IGF2 mRNA accumulation may lead to enhanced survival in a model of HCC. PMID- 12531885 TI - Ghrelin can bind to a species of high density lipoprotein associated with paraoxonase. AB - Ghrelin is a 28-residue peptide hormone that is principally released from the stomach during fasting and prior to eating. Two forms are present in human plasma: the unmodified peptide and a less abundant acylated version, in which octanoic acid is attached to the third residue, a serine, via an ester linkage. The acylated form of ghrelin acts as a ligand for the growth hormone secretagogue receptor and can stimulate the release of growth hormone from the pituitary gland. It also initiates behavioral and metabolic adaptations to fasting. Here we show that an immobilized form of ghrelin specifically binds a species of high density lipoprotein associated with the plasma esterase, paraoxonase, and clusterin. Both free ghrelin and paraoxon, a substrate for paraoxonase, can inhibit this interaction. An endogenous species of ghrelin is found to co-purify with high density lipoprotein during density gradient centrifugation and subsequent gel filtration. This interaction links the orexigenic peptide hormone ghrelin to lipid transport and metabolism. Furthermore, the interaction of the esterified hormone ghrelin with a species of HDL containing an esterase suggests a possible mechanism for the conversion of ghrelin to des-acyl ghrelin. PMID- 12531886 TI - Conformation of CCAAT/enhancer-binding protein alpha dimers varies with intranuclear location in living cells. AB - The structure of a protein defines its biochemical properties, but the impact of intracellular location and environment on protein structure remains poorly defined. CCAAT/enhancer-binding protein alpha (C/EBPalpha) is a master regulator of transcription and cellular proliferation that concentrates and is kept inactive at transcriptionally quiescent, pericentromeric regions in mouse cell nuclei. C/EBPalpha dimer structure was measured in living cells from the amounts of fluorescence energy transferred between derivatives of the green fluorescent protein attached to different C/EBPalpha domains. Comparing the levels of fluorescence resonance energy transfer at pericentromeric and nonpericentromeric regions of the nucleus indicated that the DNA binding domains of C/EBPalpha dimers were further apart and interacted more poorly at pericentromeric heterochromatin than in the more euchromatic regions of the nucleus. In contrast, the position and interactions of the transcriptional activation domains were similar throughout the nucleus. Phorbol ester treatment caused a shift in the position of the transcriptional activation domain relative to the DNA binding domain. Thus, C/EBPalpha conformation varies with intranuclear location and with cellular environment. These "fluorescence resonance energy transfer nanoscopy" techniques will be broadly applicable for associating conformational and kinetic variations to subcompartment-specific actions of C/EBPalpha or any protein in the dynamic intracellular environment. PMID- 12531884 TI - Prolactin modulates the naive B cell repertoire. AB - Prolactin is a peptide hormone produced by the anterior pituitary gland that is critical in lactation. Prolactin can also be produced by lymphocytes, and both B and T cells express prolactin receptors. These findings have suggested that prolactin has immunomodulatory functions. Studies in spontaneously autoimmune hosts have demonstrated a role for prolactin in augmenting autoreactivity. We chose to analyze prolactin effects on anti-DNA B cells in nonspontaneously autoimmune female BALB/c mice transgenic for the heavy chain of an anti-DNA antibody. Treatment with prolactin for 4 weeks induced a lupus-like phenotype with an increased number of transgene-expressing B cells, elevated serum anti-DNA antibody titers, and glomerular immunoglobulin deposits. Prolactin caused a decrease in the population of transitional B cells and an increase in mature follicular and marginal zone B cells. The DNA-reactive B cells had a follicular cell phenotype. Anti-DNA hybridomas demonstrated that prolactin alters selection of the naive B cell repertoire. The expansion and activation of anti-DNA B cells in prolactin-treated R4A-gamma2b BALB/c mice was dependent on the presence of CD4(+) T cells. Finally, treatment with prolactin was unable to break tolerance in R4A-gamma2b transgenic C57Bl/6 mice, suggesting that responsiveness of the immune system to prolactin is genetically determined. PMID- 12531887 TI - Deleted in liver cancer (DLC) 2 encodes a RhoGAP protein with growth suppressor function and is underexpressed in hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is a major malignancy in many parts of the world, especially in Asia and Africa. Loss of heterozygosity (LOH) on the long arm of chromosome 13 has been reported in HCC. In search of tumor suppressor genes in this region, here we have identified DLC2 (for deleted in liver cancer 2) at 13q12.3 encoding a novel Rho family GTPase-activating protein (GAP). DLC2 mRNA is ubiquitously expressed in normal tissues but was significantly underexpressed in 18% (8/45) of human HCCs. DLC2 is homologous to DLC1, a previously identified tumor suppressor gene at 8p22-p21.3 frequently deleted in HCC. DLC2 encodes a novel protein with a RhoGAP domain, a SAM (sterile alpha motif) domain related to p73/p63, and a lipid-binding StAR-related lipid transfer (START) domain. Biochemical analysis indicates that DLC2 protein has GAP activity specific for small GTPases RhoA and Cdc42. Expression of the GAP domain of DLC2 sufficiently inhibits the Rho-mediated formation of actin stress fibers. Introduction of human DLC2 into mouse fibroblasts suppresses Ras signaling and Ras-induced cellular transformation in a GAP-dependent manner. Taken together, our findings suggest a role for DLC2 in growth suppression and hepatocarcinogenesis. PMID- 12531888 TI - Myofibroblast differentiation by transforming growth factor-beta1 is dependent on cell adhesion and integrin signaling via focal adhesion kinase. AB - Myofibroblast differentiation and activation by transforming growth factor-beta1 (TGF-beta1) is a critical event in the pathogenesis of human fibrotic diseases, but regulatory mechanisms for this effect are unclear. In this report, we demonstrate that stable expression of the myofibroblast phenotype requires both TGF-beta1 and adhesion-dependent signals. TGF-beta1-induced myofibroblast differentiation of lung fibroblasts is blocked in non-adherent cells despite the preservation of TGF-beta receptor(s)-mediated signaling of Smad2 phosphorylation. TGF-beta1 induces tyrosine phosphorylation of focal adhesion kinase (FAK) including that of its autophosphorylation site, Tyr-397, an effect that is dependent on cell adhesion and is delayed relative to early Smad signaling. Pharmacologic inhibition of FAK or expression of kinase-deficient FAK, mutated by substituting Tyr-397 with Phe, inhibit TGF-beta1-induced alpha-smooth muscle actin expression, stress fiber formation, and cellular hypertrophy. Basal expression of alpha-smooth muscle actin is elevated in cells grown on fibronectin coated dishes but is decreased on laminin and poly-d-lysine, a non-integrin binding polypeptide. TGF-beta1 up-regulates expression of integrins and fibronectin, an effect that is associated with autophosphorylation/activation of FAK. Thus, a safer and more effective therapeutic strategy for fibrotic diseases characterized by persistent myofibroblast activation may be to target this integrin/FAK pathway while not interfering with tumor-suppressive functions of TGF-beta1/Smad signaling. PMID- 12531889 TI - Binding of filamin to the C-terminal tail of the calcitonin receptor controls recycling. AB - Many G protein-coupled receptors undergo endocytosis, but the mechanisms involved in endocytic sorting and recycling remain to be fully elucidated. We found that the G protein-coupled calcitonin receptor (CTR) undergoes tonic internalization and accumulates within the cell. Using a fluorescence loss in photobleaching assay, we classified these vesicles functionally as recycling vesicles. In a two hybrid screening, we found that the actin-binding protein filamin interacted with the C-terminal tail of the CTR. The degradation of the receptor was profoundly increased in the absence of filamin or the CTR-filamin interaction. The absence of filamin was also associated with a marked decrease in recycling of the receptor from the endosomes to the cell surface. In contrast, calcitonin-induced inhibition of spontaneous filamin proteolysis was associated with increased recycling of the receptor to the cell surface and decreased degradation of the CTR, suggesting an important role for filamin in the endocytic sorting and recycling of the internalized CTR. PMID- 12531890 TI - Degradation of phospholipid transfer protein (PLTP) and PLTP-generated pre-beta high density lipoprotein by mast cell chymase impairs high affinity efflux of cholesterol from macrophage foam cells. AB - Human atherosclerotic lesions contain mast cells filled with the neutral protease chymase. Here we studied the effect of human chymase on (i) phospholipid transfer protein (PLTP)-mediated phospholipid (PL) transfer activity, and (ii) the ability of PLTP to generate pre-beta-high density lipoprotein (HDL). Immunoblot analysis of PLTP after incubation with chymase for 6 h revealed, in addition to the original 80-kDa band, four specific proteolytic fragments of PLTP with approximate molecular masses of 70, 52, 48, and 31 kDa. This specific pattern of PLTP degradation remained stable for at least 24 h of incubation with chymase. Such proteolyzed PLTP had reduced ability (i) to transfer PL from liposome donor particles to acceptor HDL(3) particles, and (ii) to facilitate the formation of pre-beta-HDL. However, when PLTP was incubated with chymase in the presence of HDL(3), only one major cleavage product of PLTP (48 kDa) was generated, and PL transfer activity was almost fully preserved. Moreover, chymase effectively depleted the pre-beta-HDL particles generated from HDL(3) by PLTP and significantly inhibited the high affinity component of cholesterol efflux from macrophage foam cells. These results suggest that the mast cells in human atherosclerotic lesions, by secreting chymase, may prevent PLTP-dependent formation of pre-beta-HDL particles from HDL(3) and so impair the anti atherogenic function of PLTP. PMID- 12531891 TI - Impairment of human ether-a-go-go-related gene (HERG) K+ channel function by hypoglycemia and hyperglycemia. Similar phenotypes but different mechanisms. AB - Hyperglycemia and hypoglycemia both can cause prolongation of the Q-T interval and ventricular arrhythmias. Here we studied modulation of human ether-a-go-go related gene (HERG) K(+) channel, the major molecular component of delayed rectifier K(+) current responsible for cardiac repolarization, by glucose in HEK293 cells using whole-cell patch clamp techniques. We found that both hyperglycemia (extracellular glucose concentration [Glu](o) = 10 or 20 mm) and hypoglycemia ([Glu](o) = 2.5, 1, or 0 mm) impaired HERG function by reducing HERG current (I(HERG)) density, as compared with normoglycemia ([Glu](o) = 5 mm). Complete inhibition of glucose metabolism (glycolysis and oxidative phosphorylation) by 2-deoxy-d-glucose mimicked the effects of hypoglycemia, but inhibition of glycolysis or oxidative phosphorylation alone did not cause I(HERG) depression. Depletion of intracellular ATP mimicked the effects of hypoglycemia, and replacement of ATP by GTP or non-hydrolysable ATP failed to prevent the effects. Inhibition of oxidative phosphorylation by NaCN or application of antioxidants vitamin E or superoxide dismutase mimetic (Mn(III) tetrakis(4 benzoic acid) porphyrin chloride) abrogated and incubation with xanthine/xanthine oxidase mimicked the effects of hyperglycemia. Hyperglycemia or xanthine/xanthine oxidase markedly increased intracellular levels of reactive oxygen species, as measured by 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate (CM H(2)DCFDA) fluorescence dye, and this increase was prevented by NaCN, vitamin E, or Mn(III) tetrakis(4-benzoic acid) porphyrin chloride. We conclude that ATP, derived from either glycolysis or oxidative phosphorylation, is critical for normal HERG function; depression of I(HERG) in hypoglycemia results from underproduction of ATP and in hyperglycemia from overproduction of reactive oxygen species. Impairment of HERG function might contribute to Q-T prolongation caused by hypoglycemia and hyperglycemia. PMID- 12531892 TI - Elements of the glucocorticoid and retinoic acid response units are involved in cAMP-mediated expression of the PEPCK gene. AB - Although many genes are regulated by the concerted action of several hormones, hormonal signaling to gene promoters has generally been studied one hormone at a time. The phosphoenolpyruvate carboxykinase (PEPCK) gene is a case in point. Transcription of this gene is induced by glucagon (acting by the second messenger, cAMP), glucocorticoids, and retinoic acid, and it is dominantly repressed by insulin. These hormonal responses require the presence of different hormone response units (HRUs), which consist of constellations of DNA elements and associated transcription factors. These include the glucocorticoid response unit (GRU), cAMP response unit (CRU), retinoic acid response unit (RARU), and the insulin response unit. HRUs are known to have functional overlap. In particular, the cAMP response element of the CRU is also a component of the GRU. The purpose of this study was to determine whether known GRU or RARU elements or transcription factors function as components of the CRU. We show here that the glucocorticoid accessory factor binding site 1 and glucocorticoid accessory factor binding site 3 elements, which are components of both the GRU and RARU, are an important part of the CRU. Furthermore, we find that the transcription factor, chicken ovalbumin upstream promoter-transcription factor, and two coactivators, cAMP response element-binding protein-binding protein and steroid receptor coactivator-1, participate in both the cAMP and glucocorticoid responses. This provides a further illustration of how the PEPCK gene promoter integrates different hormone responses through overlapping HRUs that utilize some of the same transcription factors and coactivators. PMID- 12531893 TI - The molecular basis of differential subcellular localization of C2 domains of protein kinase C-alpha and group IVa cytosolic phospholipase A2. AB - The C2 domain is a Ca(2+)-dependent membrane-targeting module found in many cellular proteins involved in signal transduction or membrane trafficking. C2 domains are unique among membrane targeting domains in that they show a wide range of lipid selectivity for the major components of cell membranes, including phosphatidylserine and phosphatidylcholine. To understand how C2 domains show diverse lipid selectivity and how this functional diversity affects their subcellular targeting behaviors, we measured the binding of the C2 domains of group IVa cytosolic phospholipase A(2) (cPLA(2)) and protein kinase C-alpha (PKC alpha) to vesicles that model cell membranes they are targeted to, and we monitored their subcellular targeting in living cells. The surface plasmon resonance analysis indicates that the PKC-alpha C2 domain strongly prefers the cytoplasmic plasma membrane mimic to the nuclear membrane mimic due to high phosphatidylserine content in the former and that Asn(189) plays a key role in this specificity. In contrast, the cPLA(2) C2 domain has specificity for the nuclear membrane mimic over the cytoplasmic plasma membrane mimic due to high phosphatidylcholine content in the former and aromatic and hydrophobic residues in the calcium binding loops of the cPLA(2) C2 domain are important for its lipid specificity. The subcellular localization of enhanced green fluorescent protein tagged C2 domains and mutants transfected into HEK293 cells showed that the subcellular localization of the C2 domains is consistent with their lipid specificity and could be tailored by altering their in vitro lipid specificity. The relative cell membrane translocation rate of selected C2 domains was also consistent with their relative affinity for model membranes. Together, these results suggest that biophysical principles that govern the in vitro membrane binding of C2 domains can account for most of their subcellular targeting properties. PMID- 12531894 TI - Proteome analysis reveals phosphorylation of ATP synthase beta -subunit in human skeletal muscle and proteins with potential roles in type 2 diabetes. AB - Insulin resistance in skeletal muscle is a hallmark feature of type 2 diabetes. An increasing number of enzymes and metabolic pathways have been implicated in the development of insulin resistance. However, the primary cellular cause of insulin resistance remains uncertain. Proteome analysis can quantitate a large number of proteins and their post-translational modifications simultaneously and is a powerful tool to study polygenic diseases like type 2 diabetes. Using this approach on human skeletal muscle biopsies, we have identified eight potential protein markers for type 2 diabetes in the fasting state. The observed changes in protein expression indicate increased cellular stress, e.g. up-regulation of two heat shock proteins, and perturbations in ATP (re)synthesis and mitochondrial metabolism, e.g. down-regulation of ATP synthase beta-subunit and creatine kinase B, in skeletal muscle of patients with type 2 diabetes. Phosphorylation appears to play a key, potentially coordinating role for most of the proteins identified in this study. In particular, we demonstrated that the catalytic beta-subunit of ATP synthase is phosphorylated in vivo and that the levels of a down-regulated ATP synthase beta-subunit phosphoisoform in diabetic muscle correlated inversely with fasting plasma glucose levels. These data suggest a role for phosphorylation of ATP synthase beta-subunit in the regulation of ATP synthesis and that alterations in the regulation of ATP synthesis and cellular stress proteins may contribute to the pathogenesis of type 2 diabetes. PMID- 12531895 TI - Liver X receptor-dependent repression of matrix metalloproteinase-9 expression in macrophages. AB - Matrix metalloproteinases (MMPs) are zinc endopeptidases that degrade extracellular matrix (ECM) components during normal and pathogenic tissue remodeling. Inappropriate expression of these enzymes contributes to the development of vascular pathology, including atherosclerosis. MMP-9 is expressed in its active form in atherosclerotic lesions and is believed to play an important role in vascular remodeling, smooth muscle cell migration, and plaque instability. We demonstrate here that the liver X receptors (LXRs) LXRalpha and LXRbeta inhibit basal and cytokine-inducible expression of MMP-9. Treatment of murine peritoneal macrophages with the synthetic LXR agonists GW3965 or T1317 reduces MMP-9 mRNA expression and blunts its induction by pro-inflammatory stimuli including lipopolysaccharide, interleukin-1beta, and tumor necrosis factor alpha. In contrast, macrophage expression of MMP-12 and MMP-13 is not altered by LXR ligands. We further show that the ability of LXR ligands to regulate MMP-9 expression is strictly receptor-dependent and is not observed in macrophages obtained from LXRalphabeta null mice. Analysis of the 5'-flanking region of the MMP-9 gene indicates that LXR/RXR heterodimers do not bind directly to the MMP-9 promoter. Rather, activation of LXRs represses MMP-9 expression, at least in part through antagonism of the NFkappaB signaling pathway. These observations identify the regulation of macrophage MMP-9 expression as a mechanism whereby activation of LXRs may impact macrophage inflammatory responses. PMID- 12531896 TI - The cytoprotective aminothiol WR1065 activates p53 through a non-genotoxic signaling pathway involving c-Jun N-terminal kinase. AB - WR1065 is an aminothiol with selective cytoprotective effects in normal cells compared with cancer cells. In a previous study (North, S., El-Ghissassi, F., Pluquet, O., Verhaegh, G., and Hainaut, P. (2000) Oncogene 19, 1206-1214), we have shown that WR1065 activates wild-type p53 in cultured cells. Here we show that WR1065 induces p53 to accumulate through escape from proteasome-dependent degradation. This accumulation is not prevented by inhibitors of phosphatidylinositol 3-kinases and is not accompanied by phosphorylation of Ser 15, -20, or -37, which are common targets of the kinases activated in response to DNA damage. Furthermore, WR1065 activates the JNK (c-Jun N-terminal kinase), decreases complex formation between p53 and inactive JNK, and phosphorylates p53 at Thr-81, a known site of phosphorylation by JNK. A dominant negative form of JNK (JNK-APF) reduces by 50% the activation of p53 by WR1065. Thus, WR1065 activates p53 through a JNK-dependent signaling pathway. This pathway may prove useful for pharmacological modulation of p53 activity through non-genotoxic mechanisms. PMID- 12531897 TI - Regulation of p70 S6 kinase by complex formation between the Rac guanine nucleotide exchange factor (Rac-GEF) Tiam1 and the scaffold spinophilin. AB - Tiam1 is a ubiquitous guanine nucleotide exchange factor (GEF) that activates the Rac GTPase. We have shown previously that the N terminus of Tiam1 contributes to the signaling specificity of its downstream target Rac via association with IB2, a scaffold that promotes Rac activation of a p38 kinase cascade. Here we show that the N terminus of Tiam1 can influence Rac signaling specificity in a different way by interaction with spinophilin, a scaffold that binds to p70 S6 kinase, another protein regulated by Rac. In particular, spinophilin binding promotes the plasma membrane localization of Tiam1 and enhances the ability of Tiam1 to activate p70 S6 kinase. In contrast, spinophilin binding suppresses the ability of Tiam to activate Pak1, a different Rac effector. Finally, a mutant spinophilin that cannot bind to Tiam1 suppresses serum-induced p70 S6 kinase activation in cells, suggesting that a Tiam1/spinophilin complex contributes to p70 S6 kinase regulation by extracellular signals. These findings add to a growing body of evidence supporting the concept that some Rac-GEFs not only activate Rac GTPases but also participate in the selection of Rac effector by binding to particular scaffolds that complex with components of specific Rac effector pathways. PMID- 12531898 TI - The GAFa domains of rod cGMP-phosphodiesterase 6 determine the selectivity of the enzyme dimerization. AB - Retinal rod cGMP phosphodiesterase (PDE6 family) is the effector enzyme in the vertebrate visual transduction cascade. Unlike other known PDEs that form catalytic homodimers, the rod PDE6 catalytic core is a heterodimer composed of alpha and beta subunits. A system for efficient expression of rod PDE6 is not available. Therefore, to elucidate the structural basis for specific dimerization of rod PDE6, we constructed a series of chimeric proteins between PDE6alphabeta and PDE5, which contain the N-terminal GAFa/GAFb domains, or portions thereof, of the rod enzyme. These chimeras were co-expressed in Sf9 cells in various combinations as His-, myc-, or FLAG-tagged proteins. Dimerization of chimeric PDEs was assessed using gel filtration and sucrose gradient centrifugation. The composition of formed dimeric enzymes was analyzed with Western blotting and immunoprecipitation. Consistent with the selectivity of PDE6 dimerization in vivo, efficient heterodimerization was observed between the GAF regions of PDE6alpha and PDE6beta with no significant homodimerization. In addition, PDE6alpha was able to form dimers with the cone PDE6alpha' subunit. Furthermore, our analysis indicated that the PDE6 GAFa domains contain major structural determinants for the affinity and selectivity of dimerization of PDE6 catalytic subunits. The key dimerization selectivity module of PDE6 has been localized to a small segment within the GAFa domains, PDE6alpha-59-74/PDE6beta-57-72. This study provides tools for the generation of the homodimeric alphaalpha and betabeta enzymes that will allow us to address the question of functional significance of the unique heterodimerization of rod PDE6. PMID- 12531899 TI - RGS6, RGS7, RGS9, and RGS11 stimulate GTPase activity of Gi family G-proteins with differential selectivity and maximal activity. AB - Regulator of G-protein signaling (RGS) proteins are GTPase activating proteins (GAPs) of heterotrimeric G-proteins that alter the amplitude and kinetics of receptor-promoted signaling. In this study we defined the G-protein alpha-subunit selectivity of purified Sf9 cell-derived R7 proteins, a subfamily of RGS proteins (RGS6, -7, -9, and -11) containing a Ggamma-like (GGL) domain that mediates dimeric interaction with Gbeta(5). Gbeta(5)/R7 dimers stimulated steady state GTPase activity of Galpha-subunits of the G(i) family, but not of Galpha(q) or Galpha(11), when added to proteoliposomes containing M2 or M1 muscarinic receptor coupled G-protein heterotrimers. Concentration effect curves of the Gbeta(5)/R7 proteins revealed differences in potencies and efficacies toward Galpha-subunits of the G(i) family. Although all four Gbeta(5)/R7 proteins exhibited similar potencies toward Galpha(o), Gbeta(5)/RGS9 and Gbeta(5)/RGS11 were more potent GAPs of Galpha(i1), Galpha(i2), and Galpha(i3) than were Gbeta(5)/RGS6 and Gbeta(5)/RGS7. The maximal GAP activity exhibited by Gbeta(5)/RGS11 was 2- to 4 fold higher than that of Gbeta(5)/RGS7 and Gbeta(5)/RGS9, with Gbeta(5)/RGS6 exhibiting an intermediate maximal GAP activity. Moreover, the less efficacious Gbeta(5)/RGS7 and Gbeta(5)/RGS9 inhibited Gbeta(5)/RGS11-stimulated GTPase activity of Galpha(o). Therefore, R7 family RGS proteins are G(i) family selective GAPs with potentially important differences in activities. PMID- 12531900 TI - Characterization of the molecular defects in Rab27a, caused by RAB27A missense mutations found in patients with Griscelli syndrome. AB - Rab27a plays a pivotal role in the transport of melanosomes to dendrite tips of melanocytes and mutations in RAB27A, which impair melanosome transport cause the pigmentary dilution and the immune deficiency found in several patients with Griscelli syndrome (GS). Interestingly, three GS patients present single homozygous missense mutations in RAB27A, leading to W73G, L130P, and A152P transitions that affect highly conserved residues among Rab proteins. However, the functional consequences of these mutations have not been studied. In the present report, we evaluated the effect of overexpression of these mutants on melanosome, melanophilin, and myosin-Va localization in B16 melanoma cells. Then we studied several key parameters for Rab27a function, including GTP binding and interaction with melanophilin/myosin-Va complex, which links melanosomes to the actin network. Our results showed that Rab27a-L130P cannot bind GTP, does not interact with melanophilin, and consequently cannot allow melanosome transport on the actin filaments. Interestingly, Rab27a-W73G binds GTP but does not interact with melanophilin. Thus, Rab27a-W73G cannot support the actin-dependent melanosome transport. Finally, Rab27a-A152P binds both GTP and melanophilin. However, Rab27a-A152P does not allow melanosome transport and acts as a dominant negative mutant, because its overexpression, in B16 melanoma cells, mimics a GS phenotype. Hence, the interaction of Rab27a with melanophilin/myosin-Va is not sufficient to ensure a correct melanosome transport. Our results pointed to an unexpected complexity of Rab27a function and open the way to the search for new Rab27a effectors or regulators that control the transport of Rab27a-dependent vesicles. PMID- 12531902 TI - Parallel measurement of Ca2+ binding and fluorescence emission upon Ca2+ titration of recombinant skeletal muscle troponin C. Measurement of sequential calcium binding to the regulatory sites. AB - Calcium binding to chicken recombinant skeletal muscle TnC (TnC) and its mutants containing tryptophan (F29W), 5-hydroxytryptophan (F29HW), or 7-azatryptophan (F29ZW) at position 29 was measured by flow dialysis and by fluorescence. Comparative analysis of the results allowed us to determine the influence of each amino acid on the calcium binding properties of the N-terminal regulatory domain of the protein. Compared with TnC, the Ca(2+) affinity of N-terminal sites was: 1) increased 6-fold in F29W, 2) increased 3-fold in F29ZW, and 3) decreased slightly in F29HW. The Ca(2+) titration of F29ZW monitored by fluorescence displayed a bimodal curve related to sequential Ca(2+) binding to the two N terminal Ca(2+) binding sites. Single and double mutants of TnC, F29W, F29HW, and F29ZW were constructed by replacing aspartate by alanine at position 30 (site I) or 66 (site II) or both. Ca(2+) binding data showed that the Asp --> Ala mutation at position 30 impairs calcium binding to site I only, whereas the Asp --> Ala mutation at position 66 impairs calcium binding to both sites I and II. Furthermore, the Asp --> Ala mutation at position 30 eliminates the differences in Ca(2+) affinity observed for replacement of Phe at position 29 by Trp, 5 hydroxytryptophan, or 7-azatryptophan. We conclude that position 29 influences the affinity of site I and that Ca(2+) binding to site I is dependent on the previous binding of metal to site II. PMID- 12531901 TI - RICS, a novel GTPase-activating protein for Cdc42 and Rac1, is involved in the beta-catenin-N-cadherin and N-methyl-D-aspartate receptor signaling. AB - Cadherin adhesion molecules are believed to be important for synaptic plasticity. beta-Catenin, which links cadherins and the actin cytoskeleton, is a modulator of cadherin adhesion and regulates synaptic structure and function. Here we show that beta-catenin interacts with a novel GTPase-activating protein, named RICS, that acts on Cdc42 and Rac1. The RICS-beta-catenin complex was found to be associated with N-cadherin, N-methyl-d-aspartate receptors, and postsynaptic density-95, and localized to the postsynaptic density. Furthermore, the GTPase activating protein activity of RICS was inhibited by phosphorylation by Ca(2+)/calmodulin-dependent protein kinase II. These results suggest that RICS is involved in the synaptic adhesion- and N-methyl-d-aspartate-mediated organization of cytoskeletal networks and signal transduction. Thus, RICS may regulate dendritic spine morphology and strength by modulating Rho GTPases. PMID- 12531903 TI - Expression of sterol 27-hydroxylase (CYP27A1) enhances cholesterol efflux. AB - Cholesterol efflux from CHOP cells transfected with sterol 27-hydroxylase (CYP27A1) was compared with non-transfected and mock-transfected cells. Transfection caused expression of CYP27A1, formation of 27-hydroxycholesterol, and inhibition of cholesterol biosynthesis. Transfection enhanced cholesterol efflux to apolipoprotein A-I or human plasma by 2-3-fold but did not affect the efflux in the absence of acceptor. The analysis of released sterols revealed that 27-hydroxycholesterol represented only a small proportion of sterols, most of which was non-oxidized cholesterol. Time course and dose dependence studies showed that expression of CYP27A1 in CHOP cells mostly affected the efflux of the "fast" cholesterol pool, and relatively more cholesterol was released with low concentrations of an acceptor. Preincubation of non-transfected cells with exogenous 27-hydroxycholesterol (10(-9) and 10(-7) m) led to the stimulation of cholesterol efflux by 24-60%. Expression of CYP27A1 in CHOP cells did not affect ABCA1 expression and abundance of ABCA1 protein. Thus, introduction of CYP27A1 into cells stimulates cholesterol efflux and therefore may increase protection against atherosclerosis. PMID- 12531904 TI - Probing the H-protein-induced conformational change and the function of the N terminal region of Escherichia coli T-protein of the glycine cleavage system by limited proteolysis. AB - T-protein, a component of the glycine cleavage system, catalyzes a tetrahydrofolate-dependent reaction. Previously, we reported a conformational change of Escherichia coli T-protein upon interacting with E. coli H-protein (EH), showing an important role for the N-terminal region of the T-protein in the interaction. To further investigate the T-protein catalysis, the wild type (ET) and mutants were subjected to limited proteolysis. ET was favorably cleaved at Lys(81), Lys(154), Lys(288), and Lys(360) by lysylendopeptidase and the cleavages at Lys(81) and Lys(288) were strongly prevented by EH. Although ET was highly resistant to trypsinolysis, the mutant with an N-terminal 7-residue deletion (ETDelta7) was quite susceptible and instantly cleaved at Arg(16) accompanied by the rapid degradation of the resulting C-terminal fragment, indicating that the cleavage at Arg(16) is the trigger for the C-terminal fragmentation. EH showed no protection from the N-terminal cleavage, although substantial protection from the C-terminal fragmentation was observed. The replacement of Leu(6) of ET with alanine resulted in a similar sensitivity to trypsin as ETDelta7. These results suggest that the N-terminal region of ET functions as a molecular "hasp" to hold ET in the compact form required for the proper association with EH. Leu(6) seems to play a central role in the hasp function. Interestingly, Lys(360) of ET was susceptible to proteolysis even after the stabilization of the entire molecule of ET by EH, indicating its location at the surface of the ET-EH complex. Together with the buried position of Lys(81) in the complex and previous results on folate binding sites, these results suggest the formation of a folate-binding cavity via the interaction of ET with EH. The polyglutamyl tail of the folate substrate may be inserted into the bosom of the cavity leaving the pteridine ring near the entrance of the cavity in the context of the catalytic reaction. PMID- 12531905 TI - Evidence for CD4-enchanced signaling through the chemokine receptor CCR5. AB - The chemokine receptor CCR5 is constitutively associated with the T cell co receptor CD4 in plasma cell membranes, but the physiological role of this interaction has not been elucidated. Here we show that detergent-solubilized, purified CCR5 can directly associate with purified soluble fragments of the extracellular portion of CD4. We further demonstrate that the physical association of CCR5 and CD4 in membrane vesicles results in the formation of a receptor complex that exhibits macrophage inflammatory protein 1beta (MIP-1beta) binding properties that are distinct from CCR5. The affinity of the CD4-CCR5 complex for MIP-1beta was 3.5-fold lower than for CCR5, but the interaction of CD4 and CCR5 resulted in a receptor complex that exhibited enhanced G-protein signaling as compared with CCR5 alone. MIP-1beta-induced G-protein activation was further increased by simultaneous stimulation of CD4 with its natural agonist, interleukin-16. Thus, the physical association of CD4 and CCR5 results in receptor cross-talk with allosteric CD4-dependent regulation of the binding and signaling properties of CCR5. Although the precise physiological role of the CD4 effects on CCR5-mediated signaling remains unknown, one can speculate that the cross-talk is a component of mechanisms involved in the fine tuning of immune system cell responses. PMID- 12531906 TI - Importance of conserved Thr214 in domain A of the Na+,K+ -ATPase for stabilization of the phosphoryl transition state complex in E2P dephosphorylation. AB - Thr(214) of the highly conserved (214)TGES sequence in domain A of the Na(+),K(+) ATPase was replaced with alanine, and the mutant was compared functionally with the previously characterized domain A mutant Gly(263) --> Ala. Thr(214) --> Ala displayed a conspicuous 150-fold reduction of the apparent vanadate affinity for inhibition of ATPase activity, which could not simply be explained by the observed shifts of the conformational equilibria in favor of E(1) and E(1)P. The intrinsic vanadate affinity of the E(2) form and the effect on the apparent vanadate affinity of displacement of the E(1)-E(2) equilibrium were determined in a phosphorylation assay that allows the enzyme-vanadate complex to be formed under equilibrium conditions. When the E(2) form prevailed, Thr(214) --> Ala retained a reduced vanadate affinity relative to wild type, whereas the affinity of Gly(263) --> Ala became wild type-like. Thus, mutation of Thr(214) affected the intrinsic affinity of E(2) for vanadate. Furthermore, Thr(214) --> Ala showed at least a 5-fold reduced E(2)P dephosphorylation rate relative to wild type in the presence of saturating concentrations of K(+) and Mg(2+). Because vanadate is a phosphoryl transition state analog, it is proposed that defective binding of the phosphoryl transition state complex (transition state destabilization) causes the inability to catalyze E(2)P dephosphorylation properly. By contrast, the phosphorylation site in the E(1) form was unaffected in Thr(214) --> Ala. Replacement of the glutamate, Glu(216), of (214)TGES with alanine was incompatible with cell viability, indicating a very low transport activity or expression level. Our results support the hypothesis that domain A is isolated in the E(1) form, but contributes to make up the catalytic site in the E(2) and E(2)P conformations. PMID- 12531907 TI - An outer membrane enzyme that generates the 2-amino-2-deoxy-gluconate moiety of Rhizobium leguminosarum lipid A. AB - The structures of Rhizobium leguminosarum and Rhizobium etli lipid A are distinct from those found in other Gram-negative bacteria. Whereas the more typical Escherichia coli lipid A is a hexa-acylated disaccharide of glucosamine that is phosphorylated at positions 1 and 4', R. etli and R. leguminosarum lipid A consists of a mixture of structurally related species (designated A-E) that lack phosphate. A conserved distal unit, comprised of a diacylated glucosamine moiety with galacturonic acid residue at position 4' and a secondary 27 hydroxyoctacosanoyl (27-OH-C28) as part of a 2' acyloxyacyl moiety, is present in all five components. The proximal end is heterogeneous, differing in the number and lengths of acyl chains and in the identity of the sugar itself. A proximal glucosamine unit is present in B and C, but an unusual 2-amino-2-deoxy-gluconate moiety is found in D-1 and E. We now demonstrate that membranes of R. leguminosarum and R. etli can convert B to D-1 in a reaction that requires added detergent and is inhibited by EDTA. Membranes of Sinorhizobium meliloti and E. coli lack this activity. Mass spectrometry demonstrates that B is oxidized in vitro to a substance that is 16 atomic mass units larger, consistent with the formation of D-1. The oxidation of the lipid A proximal unit is also demonstrated by matrix-assisted laser desorption ionization time-of-flight mass spectrometry in the positive and negative modes using the model substrate, 1-dephospho-lipid IV(A). With this material, an additional intermediate (or by product) is detected that is tentatively identified as a lactone derivative of 1-dephospho-lipid IV(A). The enzyme, presumed to be an oxidase, is located exclusively in the outer membrane of R. leguminosarum as judged by sucrose gradient analysis. To our knowledge, an oxidase associated with the outer membranes of Gram-negative bacteria has not been reported previously. PMID- 12531909 TI - Inspiratory loading does not accelerate dystrophy in mdx mouse diaphragm: implications for regenerative therapy. AB - Since the finding that the mdx mouse diaphragm, in contrast to limb muscles, undergoes progressive degeneration analogous to that seen in Duchenne muscular dystrophy, the relationship between the workload on a muscle and the pathogenesis of dystrophy has remained controversial. We increased the work performed by the mdx mouse diaphragm in vivo by tracheal banding and evaluated the progression of dystrophic changes in that muscle. Despite the establishment of dramatically increased respiratory workload and accelerated myofiber damage documented by Evans blue dye, no change in the pace of progression of dystrophy was seen in banded animals vs. unbanded, sham-operated controls. At the completion of the study, more centrally nucleated fibers were evident in the diaphragms of banded mdx mice than in sham-operated mdx controls, indicating that myofiber regeneration increases to meet the demands of the work-induced damage. These data suggest that there is untapped regenerative capacity in dystrophin-deficient muscle and validates experimental efforts aimed at augmenting regeneration within skeletal muscle as a therapeutic strategy in the treatment of dystrophinopathies. PMID- 12531910 TI - Percent body fat via DEXA: comparison with a four-compartment model. AB - This study compared body composition by dual-energy X-ray absorptiometry (DEXA; Lunar DPX-L) with that via a four-compartment (4C; water, bone mineral mass, fat, and residual) model. Relative body fat was determined for 152 healthy adults [30.0 +/- 11.1 (SD) yr; 75.10 +/- 14.88 kg; 176.3 +/- 8.7 cm] aged from 18 to 59 yr. The 4C approach [20.7% body fat (%BF)] resulted in a significantly (P < 0.001) higher mean %BF compared with DEXA (18.9% BF), with intraindividual variations ranging from -2.6 to 7.3% BF. Linear regression and a Bland and Altman plot demonstrated the tendency for DEXA to progressively underestimate the %BF of leaner individuals compared with the criterion 4C model (4C %BF = 0.862 x DEXA %BF + 4.417; r(2) = 0.952, standard error of estimate = 1.6% BF). This bias was not attributable to variations in fat-free mass hydration but may have been due to beam-hardening errors that resulted from differences in anterior-posterior tissue thickness. PMID- 12531908 TI - Origin of the 2-amino-2-deoxy-gluconate unit in Rhizobium leguminosarum lipid A. Expression cloning of the outer membrane oxidase LpxQ. AB - An unusual feature of the lipid A from the plant endosymbionts Rhizobium etli and Rhizobium leguminosarum is the presence of a proximal sugar unit consisting of a 2-amino-2-deoxy-gluconate moiety in place of glucosamine. An outer membrane oxidase that generates the 2-amino-2-deoxy-gluconate unit from a glucosamine containing precursor is present in membranes of R. leguminosarum and R. etli but not in S. meliloti or Escherichia coli. We now report the identification of a hybrid cosmid that directs the overexpression of this activity by screening 1800 lysates of individual colonies of a R. leguminosarum 3841 genomic DNA library in the host strain R. etli CE3. Two cosmids (p1S11D and p1U12G) were identified in this manner and transferred into S. meliloti, in which they also directed the expression of oxidase activity in the absence of any chromosomal background. Subcloning and sequencing of the oxidase gene on a 6.5-kb fragment derived from the approximately 20-kb insert in p1S11D revealed that the enzyme is encoded by a gene (lpxQ) that specifies a protein of 224 amino acid residues with a putative signal sequence cleavage site at position 28. Heterologous expression of lpxQ using the T7lac promoter system in E. coli resulted in the production of catalytically active oxidase that was localized in the outer membrane. A new outer membrane protein of the size expected for LpxQ was present in this construct and was subjected to microsequencing to confirm its identity and the site of signal peptide cleavage. LpxQ expressed in E. coli generates the same products as seen in R. leguminosarum membranes. LpxQ is dependent on O(2) for activity, as demonstrated by inhibition of the reaction under strictly anaerobic conditions. An ortholog of LpxQ is present in the genome of Agrobacterium tumefaciens, as shown by heterologous expression of oxidase activity in E. coli. PMID- 12531911 TI - Citrate synthase expression and enzyme activity after endurance training in cardiac and skeletal muscles. AB - The present study was designed to examine the acute and chronic effects of endurance treadmill training on citrate synthase (CS) gene expression and enzymatic activity in rat skeletal and cardiac muscles. Adult rats were endurance trained for 8 wk on a treadmill. They were killed 1 h (T(1), n = 8) or 48 h (T(48), n = 8) after their last bout of exercise training. Eight rats were sedentary controls (C) during the training period. CS mRNA levels and enzymatic activities of the soleus and ventricle muscles were determined. Training resulted in higher CS mRNA levels in both the soleus muscles (21% increase in T(1); 18% increase in T(48), P < 0.05) and ventricle muscles (23% increase in T(1); 17% increase in T(48), P < 0.05) when compared with the C group. The CS enzyme activities were 42 (P < 0.01) and 25% (P < 0.01) greater in the soleus muscles of T(1) and T(48) groups, respectively, when compared with that of the C group. Soleus CS enzyme activity was significantly greater in the T(1) vs. T(48) groups (P < 0.05). However, no appreciable alterations in CS enzyme activities were observed in the ventricle muscles in both training groups. These findings suggest differential responses of skeletal and cardiac muscles in CS enzymatic activity but similar responses in CS gene expression at 1 and 48 h after the last session of endurance training. Moreover, our data support the existence of an acute effect of exercise on the training-induced elevation in CS activity in rat soleus but not ventricle muscles. PMID- 12531912 TI - Ventilatory response to exercise in aged runners breathing He-O2 or inspired CO2. AB - The ventilatory response to exercise below ventilatory threshold (VTh) increases with aging, whereas above VTh the ventilatory response declines only slightly. We wondered whether this same ventilatory response would be observed in older runners. We also wondered whether their ventilatory response to exercise while breathing He-O(2) or inspired CO(2) would be different. To investigate, we studied 12 seniors (63 +/- 4 yr; 10 men, 2 women) who exercised regularly (5 +/- 1 days/wk, 29 +/- 11 mi/wk, 16 +/- 6 yr). Each subject performed graded cycle ergometry to exhaustion on 3 separate days, breathing either room air, 3% inspired CO(2), or a heliox mixture (79% He and 21% O(2)). The ventilatory response to exercise below VTh was 0.35 +/- 0.06 l x min(-1) x W(-1) and above VTh was 0.66 +/- 0.10 l x min(-1) x W(-1). He-O(2) breathing increased (P < 0.05) the ventilatory response to exercise both below (0.40 +/- 0.12 l x min(-1) x W( 1)) and above VTh (0.81 +/- 0.10 l x min(-1) x W(-1)). Inspired CO(2) increased (P < 0.001) the ventilatory response to exercise only below VTh (0.44 +/- 0.10 l x min(-1) x W(-1)). The ventilatory responses to exercise with room air, He-O(2), and CO(2) breathing of these fit runners were similar to those observed earlier in older sedentary individuals. These data suggest that the ventilatory response to exercise of these senior runners is adequate to support their greater exercise capacity and that exercise training does not alter the ventilatory response to exercise with He-O(2) or inspired CO(2) breathing. PMID- 12531913 TI - Role of the autonomic nervous system in push-pull gravitational stress in anesthetized rats. AB - Tolerance to +G(z) stress is reduced by preceding exposure to -G(z) (push-pull effect). The mechanism(s) responsible for this effect are not fully understood, although the arterial baroreceptor reflexes have been implicated. We investigated the integrative response of the autonomic nervous system by studying responses to gravitational stress before and after autonomic function was inhibited by hexamethonium in 10 isoflurane-anesthetized male and female Sprague-Dawley rats. Animals were restrained supine and subjected to two rotations imposed about the x axis: 1) a control G profile consisting of rotation from 0 G(z) (+1 G(y)) to 90 degrees head-up tilt (+1 G(z)) for 10 s and 2) a push-pull G profile consisting of rotation from 0 G(z) to 90 degrees head-down tilt (-1 G(z)) for 2 s immediately preceding 10 s of +1 G(z) stress. Eight G profiles consisting of equal numbers of control and push-pull trials were imposed by using a counterbalanced design. We found that hexamethonium lowered baseline arterial pressure and abolished the push-pull effect. The lack of a push-pull effect after autonomic blockade persisted when arterial pressure was restored to baseline levels by phenylephrine infusion. Lowering baseline arterial pressure by sodium nitroprusside infusion or by hemorrhage when autonomic function was intact also abolished the push-pull effect. We conclude that intact autonomic function and a normal baseline arterial pressure are needed for expression of the push-pull effect in anesthetized rats subjected to tilting. PMID- 12531914 TI - Characterization of cardiac hypertrophy and heart failure due to volume overload in the rat. AB - Alterations in general characteristics and morphology of the heart, as well as changes in hemodynamics, myosin heavy chain isoforms, and beta-adrenoceptor responsiveness, were determined in Sprague-Dawley rats at 1, 2, 4, 8, and 16 wk after aortocaval fistula (shunt) was induced by the needle technique. Three stages of cardiac hypertrophy due to volume overload were recognized during the 16-wk period. Developing hypertrophy occurred within the first 2 wk after aortocaval shunt was induced and was characterized by a rapid increase of cardiac mass in both left and right ventricles. Compensated hypertrophy occurred between 2 and 8 wk after aortocaval shunt where normal or mild depression in hemodynamic function was observed. Decompensated hypertrophy or heart failure occurred between 8 and 16 wk after aortocaval shunt and was characterized by circulatory congestion, decreased in vivo and in vitro cardiac function, and a shift in myosin heavy chain isozyme expression. However, the positive inotropic effect of isoproterenol was augmented at all times during the 16-wk period. Characterization of beta-adrenoceptor binding in failing hearts at 16 wk revealed a significant increase in beta(1)-receptor density, whereas beta(2)-receptor density was unchanged. Consistent with this, basal adenylyl cyclase activity was significantly increased, and both isoproterenol- and forskolin-stimulated adenylyl cyclase activities were also increased. These results indicate that upregulation of beta-adrenoceptor signal transduction is a unique feature of cardiac hypertrophy and failure induced by volume overload. PMID- 12531915 TI - Plasticity in the control of breathing following sensory denervation. AB - The purpose of this manuscript is to review the results of studies on the recovery or plasticity following a denervation- or lesion-induced change in breathing. Carotid body denervation (CBD), lung denervation (LD), cervical (CDR) and thoracic (TDR) dorsal rhizotomy, dorsal spinal column lesions, and lesions at pontine, medullary, and spinal sites all chronically alter breathing. The plasticity after these is highly variable, ranging from near complete recovery of the peripheral chemoreflex in rats after CBD to minimal recovery of the Hering Breuer inflation reflex in ponies after LD. The degree of plasticity varies among the different functions of each pathway, and plasticity varies with the age of the animal when the lesion was made. In addition, plasticity after some lesions varies between species, and plasticity is greater in the awake than in the anesthetized state. Reinnervation is not a common mechanism of plasticity. There is evidence supporting two mechanisms of plasticity. One is through upregulation of an alternate sensory pathway, such as serotonin-mediated aortic chemoreception after CBD. The second is through upregulation on the efferent limb of a reflex, such as serotonin-mediated increased responsiveness of phrenic motoneurons after CDR, TDR, and spinal cord injury. Accordingly, numerous components of the ventilatory control system exhibit plasticity after denervation or lesion-induced changes in breathing; this plasticity is uniform neither in magnitude nor in underlying mechanisms. A major need in future research is to determine whether "reorganization" within the central nervous system contributes to plasticity following lesion-induced changes in breathing. PMID- 12531916 TI - The crossed phrenic phenomenon: a model for plasticity in the respiratory pathways following spinal cord injury. AB - Hemisection of the cervical spinal cord rostral to the level of the phrenic nucleus interrupts descending bulbospinal respiratory pathways, which results in a paralysis of the ipsilateral hemidiaphragm. In several mammalian species, functional recovery of the paretic hemidiaphragm can be achieved by transecting the contralateral phrenic nerve. The recovery of the paralyzed hemidiaphragm has been termed the "crossed phrenic phenomenon." The physiological basis for the crossed phrenic phenomenon is as follows: asphyxia induced by spinal hemisection and contralateral phrenicotomy increases central respiratory drive, which activates a latent crossed respiratory pathway. The uninjured, initially latent pathway mediates the hemidiaphragm recovery by descending into the spinal cord contralateral to the hemisection and then crossing the midline of the spinal cord before terminating on phrenic motoneurons ipsilateral and caudal to the hemisection. The purpose of this study is to review work conducted on the crossed phrenic phenomenon and to review closely related studies focusing particularly on the plasticity associated with the response. Because the review deals with recovery of respiratory muscles paralyzed by spinal cord injury, the clinical relevance of the reviewed studies is highlighted. PMID- 12531918 TI - Interpreting Penh in mice. PMID- 12531919 TI - Deletion of the RAG2 C terminus leads to impaired lymphoid development in mice. AB - The recombination-activating gene (RAG)1 and RAG2 proteins comprise the lymphocyte-specific components of the V(D)J recombinase and are required for the assembly of antigen-receptor variable-region genes. A mutant truncated RAG2 protein ("core" RAG2) lacking the C-terminal 144 amino acids, together with core RAG1, is able to mediate the basic biochemical steps required for V(D)J recombination in vitro and in transfected cell lines. Here we examine the effect of replacing the endogenous RAG2 locus in mice with core RAG2. These mice generate substantial numbers of B and T cells, demonstrating that the core RAG2 protein retains significant in vivo function. However, core RAG2 mice display a reduction in the total number of B and T cells, reflecting impaired lymphocyte development at the progenitor stage associated with reduced chromosomal V(D)J recombination. We discuss potential roles of the RAG2 C terminus in mediating rearrangement of endogenous antigen-receptor loci. PMID- 12531920 TI - Understanding missense mutations in the BRCA1 gene: an evolutionary approach. AB - The role of missense changes in BRCA1 in breast cancer susceptibility has been difficult to establish. We used comparative evolutionary methods to identify potential functionally important amino acid sites in exon 11 and missense changes likely to disrupt gene function, aligning sequences from 57 eutherian mammals and categorizing amino acid sites by degree of conservation. We used Bayesian phylogenetic analyses to determine relationships among orthologs and identify codons evolving under positive selection. Most conserved residues occur in a region with the highest concentration of protein-interacting domains. Rapidly evolving residues are concentrated in the RAD51-interacting domain, suggesting that selection is acting most strongly on the role of BRCA1 in DNA repair. Investigation of the functional role of missense changes in breast-cancer susceptibility should focus on 38 missense changes in conserved and 3 in rapidly evolving regions of exon 11. PMID- 12531921 TI - The C-terminal domain of myosin-like protein 1 (Mlp1p) is a docking site for heterogeneous nuclear ribonucleoproteins that are required for mRNA export. AB - For mRNA to be transported from the nucleus to the cytoplasm, it must travel from the site of transcription through the nuclear interior to the nuclear pore. Studies in Saccharomyces cerevisiae have suggested a relationship between poly(A) RNA trafficking and myosin-like protein 1 (Mlp1p), a nuclear-pore associated protein that is homologous to the mammalian Tpr (translocated promoter region) protein [Kosova, B., Pante, N., Rollenhagen, C., Podtelejnikov, A., Mann, M., Aebi, U., and Hurt, E. (2000) J. Biol. Chem. 275, 343-350]. We identified a yeast two-hybrid interaction between the C-terminal globular domain of Mlp1p and Nab2p, a shuttling heterogeneous nuclear ribonucleoprotein that is required for mRNA export. Coimmunoprecipitation confirms that Nab2p also interacts with full-length Mlp1p and in vitro binding experiments show that Nab2p binds directly to the C terminal domain of Mlp1p. In addition, our experiments reveal that the C-terminal domain of Mlp1p is both necessary and sufficient to cause accumulation of poly(A) RNA and Nab2p in the nucleus. We propose a model where Mlp1p acts as a checkpoint at the nuclear pore by interacting with export-competent ribonucleoprotein complexes through its C-terminal globular domain. This study identifies Nab2p as a heterogeneous nuclear ribonucleoprotein found in complex with Mlp1p and begins to delineate the path that mRNA travels from the chromatin to the nuclear pore. PMID- 12531922 TI - Definitive separation of graft-versus-leukemia- and graft-versus-host-specific CD4+ T cells by virtue of their receptor beta loci sequences. AB - Although graft-versus-host (GVH) disease (GVHD) is usually associated with graft versus leukemia (GVL), GVL can occur in the absence of clinical GVHD. There is evidence to suggest that GVL and GVH are mediated by different clones of T cells. The objective of this study was to identify the two types of T cells based on their receptor sequences. To this end we used irradiated nonleukemic cells from recipients as stimulator cells in a primary mixed leukocyte reaction (MLR). The activated CD4(+) donor T cells that expressed CD25 were purified by cell sorting. To prepare GVL-specific T cells, alloreactive T cells in the primary MLR were first depleted with an anti-CD25 immunotoxin. The remaining T cells had negligible alloreactivity in a secondary MLR. The allodepleted cells were then stimulated by using purified leukemia cells from the same individual as stimulator cells, and the CD25(+)-activated cells were purified by cell sorting. The GVL- and GVH-specific T cells were analyzed for their T cell receptor (TCR) clonality by using anchored RT-PCR of all the TCRbeta locus complementarity determining region 3 (CDR3) sequences. By comparing TCRbeta CDR3 sequences from transformed bacterial colonies, we were able to demonstrate that T cells mediating GVH were different from those mediating GVL in each of the eight HLA mismatched and one HLA-matched donor/recipient pairs. By using the appropriate TCRbeta CDR3-specific primers and probes, the GVH- and GVL-specific clones were monitored in a recipient undergoing an allogeneic stem cell transplant from her HLA-matched related donor. PMID- 12531923 TI - The renal pelvis: machinery that concentrates urine in the papilla. AB - Two decades ago, Bodil Schmidt-Nielsen and Bruce Graves documented the rhythmic contractions of the renal pelvis in a remarkable video, visually demonstrating how peristaltic waves empty the papilla and how the subsequent elastic recoil draws water from the collecting duct and into the tethered-open ascending vasa recta. Thus a periodic hydrostatic gradient generates an axial osmotic gradient. This review recapitulates the video and offers a link to figures and scenes digitized from the original tape. PMID- 12531924 TI - From the nucleus toward the cell periphery: a guided tour for mRNAs. AB - RNA processing, directed transport along cytoskeletal tracks, and site-specific translation of mRNA at the cell periphery are considered discrete steps in the generation of microfilament-membrane adhesion complexes. A recently identified member of the heterogeneous nuclear ribonucleoprotein family, raver1, may couple these steps and contribute to the assembly and maintenance of these structures. PMID- 12531926 TI - ATP as a signaling molecule: the exocrine focus. AB - Why and how do cells release ATP? It is not spilled energy. ATP becomes an extracellular regulator. Various cellular responses are initiated by purinergic receptors and signaling processes and are terminated by breakdown of ATP by ectonucleotidases. In epithelia, ATP regulates salt and water transport; other effects may be longer lasting. PMID- 12531927 TI - Small G proteins as novel therapeutic targets in cardiovascular medicine. AB - Small G proteins are implicated in regulation of endothelial function, smooth muscle cell contraction, proliferation, and migration, as well as cardiomyocyte hypertrophy. Targeting small G proteins and their downstream signaling could constitute promising therapeutic approaches in cardiovascular disorders such as atherosclerosis, restenosis, hypertension, vasospasm, and cardiac hypertrophy. PMID- 12531928 TI - Genesis and control of the respiratory rhythm in adult mammals. AB - The neural mechanisms responsible for respiratory rhythmogenesis in mammals were studied first in vivo in adults and subsequently in vitro in neonates. In vitro data have suggested that the pacemaker neurons are the kernel of the respiratory network. These data are reviewed, and their relevance to adults is discussed. PMID- 12531929 TI - Ubiquitin-dependent proteolysis in neurons. AB - Various studies identified the ubiquitin-proteasome system as the prime suspect in causing neurodegenerative diseases. The present review summarizes our current knowledge about the expression, regulation, and functions of this major protein degradation pathway in the brain, with particular reference to the pathogenesis of associated neurological diseases. PMID- 12531930 TI - Dysfunction of wild-type huntingtin in Huntington disease. AB - Huntingtin is the protein involved in Huntington disease (HD), an inherited neurodegenerative disease. Research activities have focused on the abnormal functions of mutant huntingtin. However, recent results indicate that wild-type huntingtin has important activities in brain neurons, suggesting that loss of these activities may play a role in HD. PMID- 12531931 TI - CFTR and bicarbonate secretion by [correction of to] epithelial cells. AB - Defective HCO(3)(-) and fluid secretion are hallmarks of the pathophysiology of the pancreas of cystic fibrosis patients. Recently, impaired HCO(3)(-) secretion has been shown in most tissues known to express the cystic fibrosis transmembrane conductance regulator (CFTR). New results suggest that CFTR plays an important role in the transcellular secretion of HCO(3)(-). PMID- 12531932 TI - Measuring motor complications in clinical trials for early Parkinson's disease. PMID- 12531933 TI - Dipoles and dementia. PMID- 12531934 TI - Idiopathic generalised epilepsies: not only for the paediatrician. PMID- 12531935 TI - Multidisciplinary rehabilitation for people with Parkinson's disease. PMID- 12531936 TI - Vasospastic amaurosis fugax. PMID- 12531937 TI - The deep tendon and the abdominal reflexes. AB - The deep tendon reflexes (and the abdominal reflexes) are important physical signs which have a special place in neurological diagnosis, particularly in early disease when they alone may be abnormal. They act as "hard" signs in situations where clinical assessment is complicated by patient anxiety, and become more useful as clinical experience develops. PMID- 12531938 TI - Internet resources for neurosurgeons and neuropathologists. AB - Neurosurgical and neuropathological resources on the internet are rapidly developing. Some excellent clinical, patient information, professional, academic, and teaching web sites are available. This review summarises the most useful online sites for neurosurgeons and neuropathologists in the United Kingdom and beyond. More general internet resources have been covered in the first article in this series. PMID- 12531939 TI - Multidisciplinary rehabilitation for people with Parkinson's disease: a randomised controlled study. AB - OBJECTIVE: To determine whether a programme of multidisciplinary rehabilitation and group support achieves sustained benefit for people with Parkinson's disease or their carers. METHODS: The study was a randomised controlled crossover trial comparing patients and carers who had received rehabilitation four months before assessment with those who had not. Patients were recruited from a neurology clinic, attended a day hospital from home weekly for six weeks using private car or hospital transport, and received group educational activities and individual rehabilitation from a multidisciplinary team. Patients were assessed at entry and at six months using a 25 item self assessment Parkinson's disease disability questionnaire, Euroqol-5d, SF-36, PDQ-39, hospital anxiety and depression scale, and timed stand-walk-sit test. Carers were assessed using the carer strain index and Euroqol-5d. RESULTS: 144 people with Parkinson's disease without severe cognitive losses and able to travel to hospital were registered (seven were duplicate registrations); 94 had assessments at baseline and six months. Repeated measures analysis of variance comparing patients at the 24 week crossover point showed that those receiving rehabilitation had a trend towards better stand-walk sit score (p = 0.093) and worse general and mental health (p = 0.002, p = 0.019). Carers of treated patients had a trend towards more strain (p = 0.086). Analysis comparing patients before and six months after treatment showed worsening in disability, quality of life, and carer strain. CONCLUSIONS: Patients with Parkinson's disease decline significantly over six months, but a short spell of multidisciplinary rehabilitation may improve mobility. Follow up treatments may be needed to maintain any benefit. PMID- 12531941 TI - Pick's disease. PMID- 12531940 TI - The relative health related quality of life of veterans with Parkinson's disease. AB - OBJECTIVES: To use databases of the US Veterans Health Administration (VHA) to describe the impact of Parkinson's disease on health related quality of life (HRQoL) of veterans; to compare the HRQoL of veterans with Parkinson's disease with that of veterans reporting eight other neurological or chronic conditions; and to estimate the unique effect of Parkinson's disease on HRQoL. METHODS: Respondents to the VHA 1999 large national health survey of veteran enrollees with a diagnosis of Parkinson's disease in VHA treatment files for the fiscal years 1997-1999 were identified by merging databases. The survey incorporated the Veterans SF-36, a well validated generic measure of HRQoL and functional status. This was used to compare patient groups. Mean physical (PCS) and mental (MCS) component summary scores were calculated for Parkinson's disease and eight other diseases by multivariable regressions that adjusted for age, sex, race, education, and 15 mental and physical co-morbid conditions that were self reported in the survey. RESULTS: Of 887 775 survey respondents, 14 530 (1.64%) had a Parkinson's disease diagnosis. Controlling for sociodemographic factors and co-morbidities, veterans with Parkinson's disease had PCS and MCS below veterans with angina/coronary heart disease, arthritis, chronic low back pain, congestive heart failure, diabetes, and stroke. Veterans with spinal cord injury reported slightly lower PCS than veterans with Parkinson's disease (32.38 v 32.72; 0.03 of 1 SD). Veterans with depression reported markedly lower MCS than veterans with Parkinson's disease (35.94 v 41.48; 0.55 of 1 SD). The unique effect of having Parkinson's disease on HRQoL was to lower PCS and MCS by 4.10 and 3.42 points (0.41 and 0.34 of 1 SD), respectively. CONCLUSIONS: The analysis quantifies the negative impact of Parkinson's disease on HRQoL, after controlling for sociodemographic factors and co-morbidities. Compared with eight other chronic conditions, Parkinson's disease imposes a relatively heavy burden on US veterans in the VHA health care system. PMID- 12531942 TI - Deep brain stimulation of the subthalamic nucleus: effectiveness in advanced Parkinson's disease patients previously reliant on apomorphine. AB - OBJECTIVES: To assess the efficacy of bilateral subthalamic nucleus (STN) deep brain stimulation (DBS) in patients with advanced Parkinson's disease previously reliant on apomorphine as their main antiparkinsonian medication. METHODS: Seven patients with motor fluctuations despite optimal medical treatment given as predominantly apomorphine infusion (n=6), or intermittent apomorphine injections (n=1) underwent bilateral STN DBS using frameless stereotactic surgery. Standard assessments of parkinsonism and motor fluctuations, using Unified Parkinson's Disease Rating Scale (UPDRS) were performed before and six months after surgery. Assessments were performed both on and off medication, and postoperative with the stimulators switched on and off. RESULTS: Bilateral STN DBS improved motor scores (UPDRS III) by 61% when off medication (p<0.05). Clinical fluctuations (UPDRS IV items 36-39) were reduced by 46.2% (p<0.05). Total daily apomorphine dose was reduced by 68.9% (p<0.05) and apomorphine infusion via a pump was no longer required in four patients. There were no operative complications. Two patients required treatment for hallucinations postoperatively but there was no significant change in mini-mental state examination. CONCLUSIONS: In patients with advanced Parkinson's disease, previously reliant on apomorphine, bilateral STN DBS is an effective treatment to reduce motor fluctuations and enable a reduction in apomorphine use. PMID- 12531943 TI - Cognitive and behavioural effects of chronic stimulation of the subthalamic nucleus in patients with Parkinson's disease. AB - OBJECTIVE: To investigate cognitive and behavioural effects of bilateral lead implants for high frequency stimulation (HFS) of the subthalamic nucleus in patients with Parkinson's disease; and to discriminate between HFS and the effects of surgical intervention on cognitive function by carrying out postoperative cognitive assessments with the stimulators turned on or off. METHODS: Motor, cognitive, behavioural, and functional assessments were undertaken in 20 patients with Parkinson's disease before implantation and then at three, six, and 12 months afterwards. Nine patients were also examined 18 months after surgery. Postoperative cognitive assessments were carried out with stimulators turned off at three and 18 months, and turned on at six and 12 months. RESULTS: Cognitive assessment showed a significant postoperative decline in performance on tasks of letter verbal fluency (across all postoperative assessments, but more pronounced at three months) and episodic verbal memory (only at three months, with stimulators off). At three, six, and 12 months after surgery, there was a significant improvement in the mini-mental state examination and in a task of executive function (modified Wisconsin card sorting test). On all postoperative assessments, there was an improvement in parkinsonian motor symptoms, quality of life, and activities of daily living while off antiparkinsonian drugs. A significant postoperative decrease in depressive and anxiety symptoms was observed across all assessments. Similar results were seen in the subgroup of nine patients with an 18 month follow up. Following implantation, three patients developed transient manic symptoms and one showed persistent psychic akinesia. CONCLUSIONS: Bilateral HFS of the subthalamic nucleus is a relatively safe procedure with respect to long term cognitive and behavioural morbidity, although individual variability in postoperative cognitive and behavioural outcome invites caution. Stimulation of the subthalamic nucleus does not per se appear to impair cognitive performance in patients with Parkinson's disease and may alleviate the postoperative decline in verbal fluency. PMID- 12531945 TI - Outcome of epilepsy surgery in focal cortical dysplasia. AB - OBJECTIVE: To describe the outcome of surgery in patients with drug resistant epilepsy and a histopathological diagnosis of focal cortical dysplasia. METHODS AND SUBJECTS: Analysis of histories and presurgical and follow up data was carried out in 53 patients with a histological diagnosis of focal cortical dysplasia. Their mean age was 24.0 years (range 5 to 46), and they included 14 children and adolescents. Mean age at seizure onset was 12.4 years (0.4 to 36) and mean seizure duration was 11.6 years (1 to 45). RESULTS: The presurgical detection rate of focal cortical dysplasia with magnetic resonance imaging (MRI) was 96%. There were 24 temporal and 29 extratemporal resections; additional multiple subpial transections were done in 12 cases to prevent spread of seizure discharges. There was a 6% rate of complications with permanent neurological deficit, but no deaths. All resected specimens were classified by neuropathological criteria as focal cortical dysplasia. Balloon cells were seen in most cases of extratemporal focal cortical dysplasia. After a mean follow up of 50 months, 38 patients (72%) were seizure-free, two (4%) had less than two seizures a year, nine (17%) had a reduction of seizure frequency of more than 75%, and four (8%) had no improvement. Seizure outcome was similar after temporal and extratemporal surgery. The patients in need of multilobar surgery had the poorest outcome. CONCLUSIONS: Circumscribed lesionectomy of focal dysplastic lesions provides seizure relief in patients with chronic drug resistant temporal and extratemporal epilepsy. There was a trend for the best seizure outcome to be in patients with early presurgical evaluation and early surgery, and in whom lesions were identified on the preoperative MRI studies. PMID- 12531946 TI - Non-convulsive status epilepticus: usefulness of clinical features in selecting patients for urgent EEG. AB - BACKGROUND: Non-convulsive status epilepticus (NCSE) is status epilepticus without obvious tonic-clonic activity. Patients with NCSE have altered mental state. An EEG is needed to confirm the diagnosis, but obtaining an EEG on every patient with altered mental state is not practical. OBJECTIVE: To determine whether clinical features could be used to predict which patients were more likely to be in NCSE and thus in need of an urgent EEG. METHODS: Over a six month period, all patients for whom an urgent EEG was ordered to identify NCSE were enrolled. Neurology residents examined the patients and filled out a questionnaire without knowledge of the EEG results. The patients were divided into two groups, NCSE and non-NCSE, depending on the EEG result. The clinical features were compared between the two groups. The sensitivity and specificity of the features were calculated. RESULTS: 48 patients were enrolled, 12 in NCSE and 36 not in NCSE. Remote risk factors for seizures, severely impaired mental state, and ocular movement abnormalities were seen significantly more often in the NCSE group. The combined sensitivity of remote risk factors for seizures and ocular movement abnormalities was 100%. CONCLUSIONS: There are certain clinical features that are more likely to be present in patients in NCSE compared with other types of encephalopathy. Either remote risk factors for seizures or ocular movement abnormalities were seen in all patients in NCSE. These features may be used to select which patients should have an urgent EEG. PMID- 12531947 TI - Idiopathic generalised epilepsy of adult onset: clinical syndromes and genetics. AB - OBJECTIVE: To study the clinical features and genetics of idiopathic generalised epilepsy (IGE) beginning in adult life. METHODS: Consecutive patients with IGE, defined as generalised seizures with spike or polyspike and wave on EEG, were studied in the setting of a first seizure clinic where an early postictal EEG record is part of the protocol. Patients were divided into two groups: "classical IGE" with onset before 20 years and inclusive of all the IGE subsyndromes recognised by the international classification; and "adult onset IGE", when seizure onset was at age 20 years or later. Seizure patterns, clinical features, and genetics of the adult onset group were examined. RESULTS: Of 121 patients with an electro-clinical diagnosis of IGE, 34 (28%) were diagnosed as adult onset IGE. The seizure patterns in these 34 cases were tonic-clonic seizures + absences (3), tonic-clonic seizures + myoclonus (6), and tonic-clonic seizures alone (25). Tonic-clonic seizures were often precipitated by alcohol or sleep deprivation. The proportion of affected first and second degree relatives did not differ between the classical and adult onset IGE groups. Twenty adult onset cases were treated with sodium valproate, four with other antiepileptic drugs, and 10 were untreated. Follow up of 32 of the 34 cases (for 31 (22) months (mean (SD)) showed that tonic-clonic seizures recurred in eight patients: five with identified provocative factors and three without. CONCLUSIONS: Adult onset IGE is a relatively frequent and benign disorder. Seizures are usually provoked and are easy to control. Patients in this age group may often be misdiagnosed as having non-lesional partial epilepsy. Early postictal EEG and sleep deprivation studies may improve the detection of these patients. Pedigree analysis suggests that adult onset IGE, like classical IGE, has a genetic aetiology. PMID- 12531948 TI - Expression of beta2 adrenoreceptors on peripheral blood mononuclear cells in patients with primary and secondary progressive multiple sclerosis: a longitudinal six month study. AB - BACKGROUND: Beta(2) adrenoreceptor expression on peripheral blood mononuclear cells is increased in progressive multiple sclerosis. This increase has been correlated with disease activity in relapsing-remitting multiple sclerosis. OBJECTIVE: To determine the beta(2) adrenoreceptor expression in primary and secondary progressive multiple sclerosis in relation to findings on magnetic resonance imaging (MRI) and clinical disease activity. METHODS: 10 patients with multiple sclerosis were studied (five with primary progressive and five with secondary progressive forms of the disease) over a period of six months. Monthly clinical and MRI assessments of the brain and spinal cord were carried out. Beta(2) adrenoreceptor expression was assessed monthly using a ligand binding assay with [(125)I]iodocyanopindolol. Expression of beta(2) adrenoceptors on peripheral blood mononuclear cells was also assessed in five normal controls over a similar period. RESULTS: The mean (SEM) value of beta(2) adrenoreceptor density for the five normal controls was 1346 (183) sites/cell, with affinity Kd of 120 (40) pM. MRI disease activity in primary progressive multiple sclerosis was reported on two occasions and on those occasions the expression of beta(2) adrenoreceptors was increased in excess of 1900 sites/cell; in the remaining 28 observations beta(2) adrenoreceptor expression was within the normal range (800 to 1900 sites/cell). In patients with secondary progressive disease, MRI disease activity was observed on 16 occasions. In these patients expression of beta(2) adrenoreceptors was increased in excess of 2000 sites/cell in all measurements except in one subject who did not show MRI activity throughout the six months period of study. The affinity of the receptors was within the normal range in all cases. CONCLUSIONS: Increased expression of beta(2) adrenoreceptors was correlated with MRI disease activity in two patients with primary progressive multiple sclerosis. In secondary progressive multiple sclerosis, increased expression of beta(2) adrenoreceptors tended not to correlate with MRI disease activity. This may reflect a persistent Th1 immune reaction in the secondary progressive form of the disease. PMID- 12531950 TI - Pyramidal tract mapping by diffusion tensor magnetic resonance imaging in multiple sclerosis: improving correlations with disability. AB - BACKGROUND: Current magnetic resonance imaging (MRI) outcome measures such as T2 lesion load correlate poorly with disability in multiple sclerosis. Diffusion tensor imaging (DTI) of the brain can provide unique information regarding the orientation and integrity of white matter tracts in vivo. OBJECTIVE: To use this information to map the pyramidal tracts of patients with multiple sclerosis, investigate the relation between burden of disease in the tracts and disability, and compare this with more global magnetic resonance estimates of disease burden. METHODS: 25 patients with relapsing-remitting multiple sclerosis and 17 healthy volunteers were studied with DTI. An algorithm was used that automatically produced anatomically plausible maps of white matter tracts. The integrity of the pyramidal tracts was assessed using relative anisotropy and a novel measure (L(t)) derived from the compounded relative anisotropy along the tracts. The methods were compared with both traditional and more recent techniques for measuring disease burden in multiple sclerosis (T2 lesion load and "whole brain" diffusion histograms). RESULTS: Relative anisotropy and L(t) were significantly lower in patients than controls (p < 0.05). Pyramidal tract L(t) in the patients correlated significantly with both expanded disability status scale (r = -0.48, p < 0.05), and to a greater degree, the pyramidal Kurtzke functional system score (KFS-p) (r = -0.75, p < 0.0001). T2 lesion load and diffusion histogram parameters did not correlate with disability. CONCLUSIONS: Tract mapping using DTI is feasible and may increase the specificity of MRI in multiple sclerosis by matching appropriate tracts with specific clinical scoring systems. These techniques may be applicable to a wide range of neurological conditions. PMID- 12531951 TI - Georges Marinescu (1864-1938). PMID- 12531952 TI - Do cognitive patterns of brain magnetic activity correlate with hippocampal atrophy in Alzheimer's disease? AB - BACKGROUND: Many reports support the clinical validity of volumetric MRI measurements in Alzheimer's disease. OBJECTIVE: To integrate functional brain imaging data derived from magnetoencephalography (MEG) and volumetric data in patients with Alzheimer's disease and in age matched controls. METHODS: MEG data were obtained in the context of a probe-letter memory task. Volumetric measurements were obtained for lateral and mesial temporal lobe regions. RESULTS: As expected, Alzheimer's disease patients showed greater hippocampal atrophy than controls bilaterally. MEG derived indices of the degree of activation in left parietal and temporal lobe areas, occurring after 400 ms from stimulus onset, correlated significantly with the relative volume of lateral and mesial temporal regions. In addition, the size of the right hippocampus accounted for a significant portion of the variance in cognitive scores independently of brain activity measures. CONCLUSIONS: These data support the view that there is a relation between hippocampal atrophy and the degree of neurophysiological activity in the left temporal lobe. PMID- 12531953 TI - Transient ischaemic attacks are associated with increased rates of global cerebral atrophy. AB - OBJECTIVES: To determine whether patients presenting with a first transient ischaemic attack (TIA) subsequently show increased rates of brain atrophy compared with age matched controls; and to assess potential risk factors for brain atrophy in this group. METHODS: 60 patients with a first, isolated TIA and 26 age and sex matched controls were recruited. None had evidence of cognitive impairment. Vascular risk factors were treated appropriately. All subjects had volumetric imaging at the start of the study and one year later, when they were clinically reassessed. TIA patients also had serial dual echo brain imaging. Rates of whole brain atrophy were calculated from the registered volumetric scans, as was the incidence of new ischaemic lesions. In the TIA group, the degree of white matter disease was assessed. Atrophy rates and blood pressure were compared between patients and controls. RESULTS: 22 patients (37%) developed new "clinically silent" infarcts during follow up. The mean (SD) annualised percentage atrophy rate in the TIA group was significantly higher than in the controls, at 0.82 (0.39)% v 0.33 (0.3)% (p < 0.0001). In the TIA group, diastolic blood pressure (p = 0.004) and white matter disease severity (p < 0.001) were correlated with cerebral atrophy rate. Increased white matter disease was found in patients in whom new ischaemic lesions developed (p < 0.001). CONCLUSIONS: Patients presenting with a first TIA have excess global brain atrophy compared with age matched controls over the subsequent year. Increased atrophy rates following a TIA may be directly or indirectly related to increasing white matter disease and diastolic hypertension. Future studies should assess whether this atrophy inevitably leads to cognitive decline, and whether aggressive treatment of risk factors for cerebrovascular disease (particularly hypertension) after a TIA can influence outcome. PMID- 12531954 TI - Intracerebral microdialysis and CSF hydrodynamics in idiopathic adult hydrocephalus syndrome. AB - BACKGROUND: In idiopathic adult hydrocephalus syndrome (IAHS), a pathophysiological model of "chronic ischaemia" caused by an arteriosclerotic process in association with a CSF hydrodynamic disturbance has been proposed. OBJECTIVE: To investigate whether CSF hydrodynamic manipulation has an impact on biochemical markers related to ischaemia, brain tissue oxygen tension (PtiO(2)), and intracranial pressure. METHODS: A microdialysis catheter, a PtiO(2) probe, and an intracerebral pressure catheter were inserted into the periventricular white matter 0-7 mm from the right frontal horn in 10 patients with IAHS. A subcutaneous microdialysis probe was used as reference. Intracranial pressure and intracerebral PtiO(2) were recorded continuously. Samples were collected for analysis between 2 and 4 pm on day 1 (baseline) and at the same time on day 2, two to four hours after a lumbar CSF hydrodynamic manipulation. The concentrations of glucose, lactate, pyruvate, and glutamate on day 1 and 2 were compared. RESULTS: After CSF drainage, there was a significant rise in the intracerebral concentration of lactate and pyruvate. The lactate to pyruvate ratio was increased and remained unchanged after drainage. There was a trend towards a lowering of glucose and glutamate. Mean intracerebral PtiO(2) was higher on day 2 than on day 1 in six of eight patients. CONCLUSIONS: There is increased glucose metabolism after CSF drainage, as expected in a situation of postischaemic recovery. These new invasive techniques are promising tools in the future study of the pathophysiological processes in IAHS. PMID- 12531955 TI - Effect of endoscopic third ventriculostomy on neuropsychological outcome in late onset idiopathic aqueduct stenosis: a prospective study. AB - OBJECTIVE: To undertake a prospective study of the long term neuropsychological outcome in patients with late onset idiopathic aqueduct stenosis (LIAS) after endoscopic third ventriculostomy. METHODS: Six patients with LIAS were evaluated pre- and postoperatively using magnetic resonance imaging (MRI) and standardised psychometric testing procedures. Endoscopic third ventriculostomy was done using standard surgical techniques. The mean long term follow up was 81.2 weeks. RESULTS: Preoperatively, all patients had cognitive impairment, four of them showing deficits in several cognitive domains. After endoscopic third ventriculostomy, all patients improved clinically and had ventricular size reduction on MRI. Postoperative neuropsychological testing showed that five patients achieved normal or near normal cognitive functions, and one improved moderately. CONCLUSIONS: Endoscopic third ventriculostomy caused a substantial improvement in the neuropsychological deficit of LIAS patients. This was also true for patients with enlarged ventricles that might be diagnosed radiologically as "arrested hydrocephalus." PMID- 12531956 TI - Complications after gamma knife radiosurgery for benign meningiomas. AB - OBJECTIVES: To analyse the results of gamma knife radiosurgery (GKS) for the treatment of intracranial meningiomas and to assess possible factors related to the outcome and complications of such treatment. METHODS: The authors retrospectively reviewed the clinical and radiological data of 179 patients (194 lesions) treated with GKS for meningiomas between May 1992 and October 2000. The mean follow up duration was 37.3 months (range 6.4 to 86.3 months). The study determined the correlation between radiosurgical outcome including imaging changes after GKS and multiple factors such as tumour location and size, patient characteristics, venous sinus status, pre-GKS degree of oedema, other treatment modalities, and radiosurgical parameters. RESULTS: The radiological control rate was 97.1%. Magnetic resonance imaging (MRI) showed complications after GKS in 35 lesions (25.0%) among the 140 lesions followed up with MRI. Complications were divided into peritumorous imaging changes (33 lesions; 23.6%) and transient cranial nerve dysfunction (two lesions; 1.4%). Radiation induced imaging changes were seen mostly in convexity, parasagittal, and falx meningiomas that were deeply embedded in the cortex. About 60% of these were asymptomatic and the overall rate of symptomatic imaging changes was 9.3%. Neurological deficit related to imaging changes developed in only three patients, and all the symptoms were transient. CONCLUSION: GKS for intracranial meningiomas seems to be a safe and effective treatment. However, meningiomas of the convexity, parasagittal region, or falx cerebri have a higher incidence of peritumorous imaging changes after GKS than those of the skull base. Therefore, the use of GKS needs to be considered very cautiously in cerebral hemispheric meningiomas, taking into consideration patient age and general condition, tumour size and location, pattern of cortical embedding, relation between the tumour and venous sinuses, presenting symptoms, and patient preference. PMID- 12531957 TI - Quantitative analysis of continuous intracranial pressure recordings in symptomatic patients with extracranial shunts. AB - OBJECTIVES: To explore the outcome of management of possible shunt related symptoms using intracranial pressure (ICP) monitoring, and to identify potential methodological limitations with the current strategies of ICP assessment. METHODS: The distribution of persistent symptoms related to extracranial shunt treatment was compared before and after management of shunt failure in 69 consecutive hydrocephalic cases. Management was heavily based on ICP monitoring (calculation of mean ICP and visual determination of plateau waves). After the end of patient management, all ICP curves were re-evaluated using a quantitative method and software (Sensometrics pressure analyser). The ICP curves were presented as a matrix of numbers of ICP elevations (20 to 35 mm Hg) or depressions (-10 to -5 mm Hg) of different durations (0.5, 1, or 5 minutes). The numbers of ICP elevations/depressions standardised to 10 hours recording time were calculated to allow comparisons of ICP between individuals. RESULTS: After ICP monitoring and management of the putative shunt related symptoms, the symptoms remained unchanged in as many as 58% of the cases, with the highest percentages in those patients with ICP considered normal or too low at the time of ICP monitoring. The quantitative analysis revealed a high frequency of ICP elevations (20 to 35 mm Hg lasting 0.5 to 1 minute) and ICP depressions (-10 to 5 mm Hg lasting 0.5, 1, or 5 minutes), particularly in patients with ICP considered normal. CONCLUSIONS: The value of continuous ICP monitoring with ICP analysis using current criteria appears doubtful in the management of possible shunt related symptoms. This may reflect limitations in the strategies of ICP analysis. Calculation of the exact numbers of ICP elevations and depressions may provide a more accurate description of the ICP profile. PMID- 12531958 TI - Lay versus expert interviewers for the diagnosis of migraine in a large sample of elderly people. AB - OBJECTIVE: To determine the agreement between lay interviewers and experts in the diagnosis of migraine by questionnaire. SUBJECTS: A population based sample of 1188 individuals aged 64 to 73 years. METHODS: Participants who declared that they had recurrent headaches (n = 238) answered a structured questionnaire by lay interviewers with special training in migraine. A migraine expert subsequently interviewed all the headache sufferers using the same questionnaire. Migraine was defined according to the International Headache Society criteria. RESULTS: In comparison with the expert, the diagnosis derived by the lay interviewers had high values for specificity (97%) and positive predictive value (86%), and a low sensitivity (50%) and negative predictive value (57%). Agreement between the expert and the lay interviewers was low, with a kappa value of 0.36 (95% confidence interval 0.26 to 0.47). The most serious discrepancies concerned the duration of attacks, the worsening of headaches by physical activity, the presence of nausea or vomiting, and the unilaterality of headaches. As a result, the lifetime prevalence of migraine headaches was greatly underestimated by lay interviewers (6.5%) in comparison with the expert (11.1%). CONCLUSIONS: A low level of agreement between lay interviewers and a headache expert in the diagnosis of migraine headaches by structured questionnaire may result in a substantial underestimation of migraine prevalence. PMID- 12531959 TI - Deriving summary indices of health status from the Amyotrophic Lateral Sclerosis Assessment Questionnaires (ALSAQ-40 and ALSAQ-5). AB - OBJECTIVES: To use statistical procedures, operationalising what is known as item response theory (IRT), to assess the unidimensionality of the 40 item Amyotrophic Lateral Sclerosis Assessment Questionnaire, and consequently to develop a single index figure from the measure. A secondary objective is to compare scores gained on the ALSAQ-40 with a five item short form (the ALSAQ-5). METHODS: Postal survey of patients diagnosed with motor neurone disease (MND) on the MND Associations database. Copies of the ALSAQ-40 and, nested within it, the ALSAQ-5 were completed on two occasions. At time one, the survey contained the ALSAQ-40 and demographic questions. In addition, patients were asked to indicate if they were willing to take part in the follow up. Those who agreed to do so were sent another copy of the questionnaire after a period of three months. Respondents were also asked to indicate how much change they had experienced since baseline on each of the five domains of the questionnaire. Rasch analysis, a form of IRT methodology, was used to determine if the 40 items in the ALSAQ-40 tapped an underlying "latent trait", and were consequently measuring a unidimensional construct. The results from the ALSAQ-40 single index were then compared with those gained from the ALSAQ-5. RESULTS: Analyses indicated that, at both baseline and follow up, all items on the ALSAQ-40 fitted the Rasch model. Consequently the 40 items were summed to create a single index. Results on this instrument were compared with those gained by summing the five items of the ALSAQ-5. Results on the instruments were found to be highly correlated. CONCLUSIONS: Evidence from the analyses suggests that 40 item ALSAQ does contain a unidimensional scale, and can, therefore be summed to create a single index. Furthermore the ALSAQ-5 closely replicates the results of the patient measure. PMID- 12531960 TI - Visual prognosis after indirect traumatic optic neuropathy. AB - OBJECTIVE: To investigate a possible correlation between final visual acuity and the presence at baseline of various systemic and local (orbital/ocular) signs in patients affected by indirect traumatic optic neuropathy. METHODS: 35 cases of traumatic optic neuropathy were examined retrospectively and 13 variables were tested. Univariate analysis with "no recovery of visual acuity" as the primary outcome was performed. Relative risk (RR) and 95% confidence intervals (CI) were calculated. Fisher's exact test was used for two variables to test differences between proportions. RESULTS: Four variables showed a significantly increased risk for no recovery of visual acuity: presence of blood within the posterior ethmoidal cells (RR = 2.25, 95% CI 1.25 to 4.04); age over 40 years (RR = 1.79, 1.07 to 2.99); loss of consciousness associated with traumatic optic neuropathy (RR = 2.21, 1.17 to 4.16); and absence of recovery after 48 hours of steroid treatment (p < 0.01, Fisher's exact test). Recovery documented at the first follow up visit after treatment was significantly associated with recovery at the last follow up visit (p < 0.01, Fisher's exact test). CONCLUSIONS: These four negative prognostic signs in patients affected by traumatic optic neuropathy may be useful in predicting the visual outcome in patients developing visual loss after head trauma and in deciding on the need for surgical treatment. PMID- 12531961 TI - "CADASIL coma": an underdiagnosed acute encephalopathy. AB - The main clinical features of CADASIL (cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy) are stroke, dementia, and migraine. A reversible acute encephalopathy was the principal presentation in six of 70 patients in a British prevalence study. The episodes lasted seven to 14 days, presenting with fever, acute confusion, coma, and fits; there was full recovery but in two cases identical episodes recurred some years later. All patients had a previous history of migraine with aura and were originally misdiagnosed as viral encephalitis. CADASIL should be considered in acute unexplained encephalopathies. MRI white matter changes, previous migraine with aura, and a family history of stroke and dementia may be useful pointers to the diagnosis. PMID- 12531962 TI - Dissociation of affective modulation of recollective and perceptual experience following amygdala damage. AB - It has been suggested that similar neural mechanisms may underlie the affective modulation of both recollective and perceptual experience. A case is reported of a patient who has bilateral amygdala damage and marked impairment in the perception of emotion, particularly fear. The patient DR and 10 healthy control subjects (matched for school leaving age, intelligence quotient, and non emotional memory performance) were shown a series of slides accompanied by an emotionally arousing narrative. One week later DR and the controls were given a surprise memory test for this material. In addition, they completed a verbal memory test using emotionally arousing stimuli. Both DR and the healthy control subjects showed a normative pattern of enhanced memory for emotional material. On the basis of these results and the previously demonstrated impairment of perception of emotion in this patient, it is concluded that different neural mechanisms may underlie affective modulation of recollective and perceptual experience. PMID- 12531964 TI - Ataxic arm movements after thalamotomy for Parkinsonian tremor. AB - Voluntary finger-nose movements of the arm were analysed in six patients undergoing stereotaxic nucleus ventralis intermedius thalamotomy for relief of severe Parkinsonian tremor. In all cases thalamotomy acutely abolished tremor in the contralateral arm. In the early postoperative phase, ataxia of the arm contralateral to the operated side was also seen. Ataxia was transient, lasting between 7 and 21 days postoperatively. This observation suggests that a lesion of the Vim nucleus interrupts cerebellar input to the thalamus, and supports the concept that abnormal cerebellar activity is an important contributor to the generation of tremor in Parkinson's disease. PMID- 12531963 TI - Development of selective verbal memory impairment secondary to a left thalamic infarct: a longitudinal case study. AB - A 68 year old man suffered an acute dysphasic episode with persistent memory disturbance while taking part as a control in a longitudinal magnetic resonance imaging (MRI) study. A small new left thalamic infarct involving the mamillo thalamic tract could be demonstrated on volumetric MRI, coinciding with the development of a selective verbal memory impairment. This suggests that lateralisation of cognitive processing of visual and verbal material exists at the thalamic as well as the cortical level. High resolution volumetric MRI may be helpful in demonstrating small subcortical infarcts that may not be seen using computed tomography or conventional MRI. PMID- 12531965 TI - Lack of chromosomal imbalances in adamantinomatous and papillary craniopharyngiomas. AB - Craniopharyngiomas are among the most common paediatric tumours and are thought to arise from embryonic remnants of Rathke's pouch. The molecular mechanisms involved in their formation remain elusive and little is known about chromosomal imbalances that could suggest the locations of tumour suppressor or proto oncogenes involved in the pathogenesis. The paucity of published data on the molecular basis of such tumours prompted this investigation of 20 adamantinomatous and nine papillary craniopharyngiomas for genetic abnormalities by comparative genomic hybridisation (CGH). CGH revealed no DNA copy number changes in any of the 29 primary craniopharyngiomas, regardless of their histological subtype. These data suggest that chromosomal imbalances are a rare event in both adamantinomatous and papillary craniopharyngiomas. PMID- 12531966 TI - Relapsing herpes simplex encephalitis: pathological confirmation of viral reactivation. AB - This case is reported to raise awareness of herpes simplex encephalitis as a persisting brain disorder. A 66 year old immunocompetent man developed status epilepticus and died of pneumonia in the course of progressive hemiparesis, cognitive decline, and atrophy of the brain over a five year period after herpes simplex encephalitis. In addition to a completely destroyed left temporal lobe, necropsy revealed active encephalitis consisting of necrosis and lymphocyte infiltration with a large number of intranuclear inclusions in the neurones and glial cells in the markedly oedematous parenchyma of the right frontal and parietal lobes. Herpes simplex virus type 1 (HSV-1) antigen was detected by immunohistochemistry, HSV-1 DNA by in situ hybridisation, and herpes simplex virus nucleocapsids by electronmicroscopy. These clinical and pathological findings suggest that direct viral reactivation might result in a relapse of herpes simplex encephalitis, causing progressive clinical deterioration associated with the persistence of HSV-1 in the brain. This is the first case report demonstrating HSV-1 antigen, HSV-1 DNA, and herpes simplex virus nucleocapsids in a case of relapsing herpes simplex encephalitis. PMID- 12531967 TI - Does sex influence age at onset in cranial-cervical and upper limb dystonia? AB - The relation between age at dystonia onset and sex was investigated in 264 patients with cranial-cervical dystonia and 56 patients with upper limb dystonia. In cranial-cervical dystonia, women had a significantly greater age at the onset of dystonia than men. The association was independent of duration of disease and distance of referral, but it was no longer detectable after adjustment for educational level. In upper limb dystonia, men and women did not differ for age at dystonia onset, duration of disease, education level, or distance of referral. A significant inverse association between age at the onset of dystonia and education was observed in both cranial-cervical dystonia and upper limb dystonia series. PMID- 12531969 TI - Colonic transit time and rectoanal videomanometry in Parkinson's disease. AB - BACKGROUND: Constipation is a prominent lower gastrointestinal tract dysfunction that occurs frequently in Parkinson's disease (PD). OBJECTIVE: To investigate colonic transport and dynamic rectoanal behaviour during filling and defecation in patients with PD. METHODS: Colonic transit time (CTT) and rectoanal videomanometry analyses were performed in 12 patients with PD (10 men and 2 women; mean age, 68 years, mean duration of disease, five years; mean Hoehn and Yahr grade, 3; decreased stool frequency (<3 times a week) in six, difficulty in stool expulsion in eight) and 10 age matched normal control subjects (7 men and 3 women; mean age, 62 years; decreased stool frequency in two, difficulty in stool expulsion in two). RESULTS: In the PD patients, CTT was significantly prolonged in the rectosigmoid segment (p<0.05) and total colon (p<0.01) compared with the control subjects. At the resting state, anal closure and squeeze pressures of PD patients were lower than those in control subjects, though not statistically significant. However, the PD patients showed a smaller increase in abdominal pressure on coughing (p<0.01) and straining (p<0.01). The sphincter motor unit potentials of the patients were normal. During filling, PD patients showed normal rectal volumes at first sensation and maximum desire to defecate, and normal rectal compliance. However, they showed smaller amplitude in phasic rectal contraction (p<0.05), which was accompanied by an increase in anal pressure that normally decreased, together with leaking in two patients. During defecation, most PD patients could not defecate completely with larger post-defecation residuals (p<0.01). PD patients had weak abdominal strain and smaller rectal contraction on defecation than those in control subjects, though these differences were not statistically significant. However, the PD patients had larger anal contraction on defecation (p<0.05), evidence of paradoxical sphincter contraction on defecation (PSD). CONCLUSIONS: Slow colonic transit, decreased phasic rectal contraction, weak abdominal strain, and PSD were all features in our PD patients with frequent constipation. PMID- 12531970 TI - Two cases of quadriparesis following anterior cervical discectomy, with normal perioperative somatosensory evoked potentials. AB - Two cases illustrate an uncommon failure of perioperative somatosensory evoked potential (SEP) monitoring to detect iatrogenic lesions causing temporary quadriparesis during straightforward cervical surgery. In both cases, anterior cervical discectomy at one or two levels was undertaken with bone graft and titanium implants, and cortical SEP were monitored to alternate stimulation of the left and right median or ulnar nerves. The SEP showed only minor changes during surgery, not considered pathologically significant, and were normal when recorded postoperatively. Both patients, however, experienced marked postoperative limb weakness or paralysis. Motor evoked potentials (MEP) recorded postoperatively to transcranial magnetic stimulation were absent. The clinical motor deficits resolved over the ensuing months. In spite of the normally low incidence of "false negatives," in these two cases SEP monitoring failed to detect a iatrogenic lesion causing moderate to severe, though temporary, motor impairment. Monitoring of MEP may be considered as an alternative to SEP during anterior cervical procedures, while combined monitoring of SEP and MEP may be the ideal. PMID- 12531971 TI - Cerebral salt wasting syndrome in meningoencephalitis: a case report. PMID- 12531972 TI - Moderate hypoglycaemia obliterates working memory in humans with and without insulin treated diabetes. PMID- 12531973 TI - A case of familial inverse Marcus Gunn phenomenon. PMID- 12531974 TI - Parainfectious opsoclonus-myoclonus syndrome: high dose intravenous immunoglobulins are effective. PMID- 12531975 TI - Central and peripheral fusiform aneurysms six years after left atrial myxoma resection. PMID- 12531976 TI - Anti-titin antibodies are not associated with a specific thymoma histology. PMID- 12531977 TI - Transoesophageal echocardiography: which stroke patients benefit most from this investigation? PMID- 12531978 TI - Velocity dependent passive muscle stiffness. PMID- 12531979 TI - "Sentinel" or "early warning" bleed. PMID- 12531980 TI - The policy drought on climate change. PMID- 12531981 TI - Gene therapy. Second child in French trial is found to have leukemia. PMID- 12531982 TI - Science and security. Researchers urged to self-censor sensitive data. PMID- 12531983 TI - Chemical engineering. Chemists concoct quick-change surface. PMID- 12531984 TI - Physics. Confirmation of gravity's speed? Not so fast. PMID- 12531985 TI - Stem cell research. Same results, different interpretations. PMID- 12531986 TI - Evolutionary biology. On ant farm, a threesome coevolves. PMID- 12531987 TI - Planetary exploration. Scientists pick two sweet spots for Rovers on Mars. PMID- 12531988 TI - Scholarly conduct. Skeptical Environmentalist labeled 'dishonest'. PMID- 12531989 TI - Toxicology. Academy panel mulls ethics of human pesticide experiments. PMID- 12531990 TI - Evolutionary biology. Uphill dash may have led to flight. PMID- 12531992 TI - University-industry collaboration. Berkeley review dismisses critics' fears. PMID- 12531991 TI - University-industry collaboration. Last of the big-time spenders? PMID- 12531993 TI - Neuroscience. Deconstructing schizophrenia. PMID- 12531994 TI - Neuroscience. White matter's the matter. PMID- 12531996 TI - American Geophysical Union. Volcanic blasts favor El Nino warmings. PMID- 12531995 TI - American Geophysical Union. A new recipe for cooking up a 'mini solar system'. PMID- 12531997 TI - American Geophysical Union. Another way to take the ocean's pulse. PMID- 12531998 TI - Renewable energy. Norway goes with the flow to light up its nights. PMID- 12531999 TI - Comment on "Global biodiversity, biochemical kinetics, and the energetic equivalence rule". PMID- 12532001 TI - Medicine. What are the right targets for psychopharmacology? PMID- 12532002 TI - Plant biology. Seeing red. PMID- 12532003 TI - Parasitology. A game of cat and mouth. PMID- 12532004 TI - Geomorphology. On patterned ground. PMID- 12532005 TI - Development. What makes an embryo stick? PMID- 12532006 TI - Surface science. Waltzing with O2. PMID- 12532007 TI - Photonic crystal fibers. AB - Photonic crystal fibers guide light by corralling it within a periodic array of microscopic air holes that run along the entire fiber length. Largely through their ability to overcome the limitations of conventional fiber optics-for example, by permitting low-loss guidance of light in a hollow core-these fibers are proving to have a multitude of important technological and scientific applications spanning many disciplines. The result has been a renaissance of interest in optical fibers and their uses. PMID- 12532009 TI - Radio emission from an ultraluminous x-ray source. AB - The physical nature of ultraluminous x-ray sources is uncertain. Stellar-mass black holes with beamed radiation and intermediate black holes with isotropic radiation are two plausible explanations. We discovered radio emission from an ultraluminous x-ray source in the dwarf irregular galaxy NGC 5408. The x-ray, radio, and optical fluxes as well as the x-ray spectral shape are consistent with beamed relativistic jet emission from an accreting stellar black hole. If confirmed, this would suggest that the ultraluminous x-ray sources may be stellar mass rather than intermediate-mass black holes. However, interpretation of the source as a jet-producing intermediate-mass black hole cannot be ruled out at this time. PMID- 12532008 TI - Insulin staining of ES cell progeny from insulin uptake. PMID- 12532010 TI - Cerenkov radiation in photonic crystals. AB - In a conventional material, the coherent Cerenkov radiation due to a moving charged particle is associated with a velocity threshold, a forward-pointing radiation cone, and a forward direction of emission. We describe different behavior for the Cerenkov radiation in a photonic crystal. In particular, this radiation is intrinsically coupled with transition radiation and is observable without any threshold. Within one particle-velocity range, we found a radiation pattern with a backward-pointing radiation cone. In another velocity range, backward-propagating Cerenkov radiation can be expected. Potential applications include velocity-sensitive particle detection and radiation generation at selectable frequencies. PMID- 12532011 TI - A reversibly switching surface. AB - We report the design of surfaces that exhibit dynamic changes in interfacial properties, such as wettability, in response to an electrical potential. The change in wetting behavior was caused by surface-confined, single-layered molecules undergoing conformational transitions between a hydrophilic and a moderately hydrophobic state. Reversible conformational transitions were confirmed at a molecular level with the use of sum-frequency generation spectroscopy and at a macroscopic level with the use of contact angle measurements. This type of surface design enables amplification of molecular level conformational transitions to macroscopic changes in surface properties without altering the chemical identity of the surface. Such reversibly switching surfaces may open previously unknown opportunities in interfacial engineering. PMID- 12532012 TI - Spatiotemporal coherent control of lattice vibrational waves. AB - We achieved automated optical control over coherent lattice responses that were both time- and position-dependent across macroscopic length scales. In our experiments, spatiotemporal femtosecond pulse shaping was used to generate excitation light fields that were directed toward distinct regions of crystalline samples, producing terahertz-frequency lattice vibrational waves that emanated outward from their multiple origins at lightlike speeds. Interferences among the waves resulted in fully specified far-field responses, including tilted, focusing, or amplified wavefronts. Generation and coherent amplification of terahertz traveling waves and terahertz phased-array generation also were demonstrated. PMID- 12532013 TI - Self-organization of sorted patterned ground. AB - Striking circular, labyrinthine, polygonal, and striped patterns of stones and soil self-organize in many polar and high alpine environments. These forms emerge because freeze-thaw cycles drive an interplay between two feedback mechanisms. First, formation of ice lenses in freezing soil sorts stones and soil by displacing soil toward soil-rich domains and stones toward stone-rich domains. Second, stones are transported along the axis of elongate stone domains, which are squeezed and confined as freezing soil domains expand. In a numerical model implementing these feedbacks, circles, labyrinths, and islands form when sorting dominates; polygonal networks form when stone domain squeezing and confinement dominate; and stripes form as hillslope gradient is increased. PMID- 12532014 TI - The global morphology of wave Poynting flux: powering the aurora. AB - Large-scale, electric currents flowing along magnetic field lines into the polar regions of Earth are thought to be the main contributors of the energy that powers the ionospheric aurora. However, we have found evidence for global contributions from electromagnetic waves (Alfven waves). Data that were collected from the Polar spacecraft over the course of 1 year show that the flow of wave electromagnetic energy at altitudes of 25,000 to 38,000 kilometers delineates the statistical auroral oval. The Poynting flux of individual events distributed along the auroral oval was larger than 5 ergs per square centimeter per second, which is sufficient to power auroral acceleration processes. This evidence suggests that in addition to magnetic field-aligned currents, the dayside and nightside aurora is globally powered by the energy flow of these high-altitude Alfven waves. PMID- 12532015 TI - Ancient tripartite coevolution in the attine ant-microbe symbiosis. AB - The symbiosis between fungus-growing ants and the fungi they cultivate for food has been shaped by 50 million years of coevolution. Phylogenetic analyses indicate that this long coevolutionary history includes a third symbiont lineage: specialized microfungal parasites of the ants' fungus gardens. At ancient levels, the phylogenies of the three symbionts are perfectly congruent, revealing that the ant-microbe symbiosis is the product of tripartite coevolution between the farming ants, their cultivars, and the garden parasites. At recent phylogenetic levels, coevolution has been punctuated by occasional host-switching by the parasite, thus intensifying continuous coadaptation between symbionts in a tripartite arms race. PMID- 12532016 TI - Collapse and conservation of shark populations in the Northwest Atlantic. AB - Overexploitation threatens the future of many large vertebrates. In the ocean, tunas and sea turtles are current conservation concerns because of this intense pressure. The status of most shark species, in contrast, remains uncertain. Using the largest data set in the Northwest Atlantic, we show rapid large declines in large coastal and oceanic shark populations. Scalloped hammerhead, white, and thresher sharks are each estimated to have declined by over 75% in the past 15 years. Closed-area models highlight priority areas for shark conservation, and the need to consider effort reallocation and site selection if marine reserves are to benefit multiple threatened species. PMID- 12532017 TI - Selective trafficking of non-cell-autonomous proteins mediated by NtNCAPP1. AB - In plants, cell-to-cell communication is mediated by plasmodesmata and involves the trafficking of non-cell-autonomous proteins (NCAPs). A component in this pathway, Nicotiana tabacum NON-CELL-AUTONOMOUS PATHWAY PROTEIN1 (NtNCAPP1), was affinity purified and cloned. Protein overlay assays and in vivo studies showed that NtNCAPP1 is located on the endoplasmic reticulum at the cell periphery and displays specificity in its interaction with NCAPs. Deletion of the NtNCAPP1 amino-terminal transmembrane domain produced a dominant-negative mutant that blocked the trafficking of specific NCAPs. Transgenic tobacco plants expressing this mutant form of NtNCAPP1 and plants in which the NtNCAPP1 gene was silenced were compromised in their ability to regulate leaf and floral development. These results support a model in which NCAP delivery to plasmodesmata is both selective and regulated. PMID- 12532018 TI - Role of anthocyanidin reductase, encoded by BANYULS in plant flavonoid biosynthesis. AB - Condensed tannins (CTs) are flavonoid oligomers, many of which have beneficial effects on animal and human health. The flavanol (-)-epicatechin is a component of many CTs and contributes to flavor and astringency in tea and wine. We show that the BANYULS (BAN) genes from Arabidopsis thaliana and Medicago truncatula encode anthocyanidin reductase, which converts anthocyanidins to their corresponding 2,3-cis-flavan-3-ols. Ectopic expression of BAN in tobacco flower petals and Arabidopsis leaves results in loss of anthocyanins and accumulation of CTs. PMID- 12532019 TI - Hypaxial motor patterns and the function of epipubic bones in primitive mammals. AB - Since the first description of epipubic bones in 1698, their functions and those of the associated abdominal muscles of monotremes and marsupial mammals have remained unresolved. We show that each epipubic bone is part of a kinetic linkage extending from the femur, by way of the pectineus muscle, to the epipubic bone, through the pyramidalis and rectus abdominis muscles on one side of the abdomen, and through the contralateral external and internal oblique muscles to the vertebrae and ribs of the opposite side. This muscle series is activated synchronously as the femur and contralateral forelimb are retracted during the stance phase in locomotion. The epipubic bone acts as a lever that is retracted (depressed) to stiffen the trunk between the diagonal limbs that support the body during each step. This cross-couplet kinetic linkage and the stiffening function of the epipubic bone appear to be the primitive conditions for mammals. PMID- 12532020 TI - Wing-assisted incline running and the evolution of flight. AB - Flapping wings of galliform birds are routinely used to produce aerodynamic forces oriented toward the substrate to enhance hindlimb traction. Here, I document this behavior in natural and laboratory settings. Adult birds fully capable of aerial flight preferentially employ wing-assisted incline running (WAIR), rather than flying, to reach elevated refuges (such as cliffs, trees, and boulders). From the day of hatching and before attaining sustained aerial flight, developing ground birds use WAIR to enhance their locomotor performance through improved foot traction, ultimately permitting vertical running. WAIR provides insight from behaviors observable in living birds into the possible role of incipient wings in feathered theropod dinosaurs and offers a previously unstudied explanation for the evolution of avian flight. PMID- 12532021 TI - Trophoblast L-selectin-mediated adhesion at the maternal-fetal interface. AB - Trophoblast adhesion to the uterine wall is the requisite first step of implantation and, subsequently, placentation. At the maternal-fetal interface, we investigated the expression of selectin adhesion systems that enable leukocyte capture from the bloodstream. On the maternal side, human uterine epithelial cells up-regulated selectin oligosaccharide-based ligands during the window of receptivity. On the fetal side, human trophoblasts expressed L-selectin. This ligand-receptor system was functional, because beads coated with the selectin ligand 6-sulfo sLe(x) bound to trophoblasts, and trophoblasts bound to ligand expressing uterine luminal epithelium in tissue sections. These results suggest that trophoblast L-selectin mediates interactions with the uterus and that this adhesion mechanism may be critical to establishing human pregnancy. PMID- 12532022 TI - Recent expansion of Toxoplasma through enhanced oral transmission. AB - The global predominance of three clonal Toxoplasma gondii lineages suggests that they are endowed with an exceptional trait responsible for their current parasitism of nearly all warm-blooded vertebrates. Genetic polymorphism analyses indicate that these clonal lineages emerged within the last 10,000 years after a single genetic cross. Comparison with ancient strains (approximately 1 million years) suggests that the success of the clonal lineages resulted from the concurrent acquisition of direct oral infectivity. This key adaptation circumvented sexual recombination, simultaneously promoting transmission through successive hosts, hence leading to clonal expansion. Thus, changes in complex life cycles can occur rapidly and can profoundly influence pathogenicity. PMID- 12532023 TI - Contributions of the visual ventral pathway to long-range apparent motion. AB - Objects displaced intermittently across the visual field will nonetheless give an illusion of continuous motion [called apparent motion (AM)] under many common conditions. It is believed that form perception is of minor importance in determining AM, and that AM is mediated by motion-sensitive areas in the "where" pathway of the cortex. However, form and motion typically interact in specific ways when natural objects move through the environment. We used functional magnetic resonance imaging to measure cortical activation to long-range AM, compared to short-range AM and flicker, while we varied stability of structural differences between forms. Long-range AM activated the anterior-temporal lobe in the visual ventral pathway, and the response varied according to the form stability. The results suggest that long-range AM is associated with neural systems for form perception. PMID- 12532024 TI - Toll pathway-dependent blockade of CD4+CD25+ T cell-mediated suppression by dendritic cells. AB - Toll-like receptors (TLRs) control activation of adaptive immune responses by antigen-presenting cells (APCs). However, initiation of adaptive immune responses is also controlled by regulatory T cells (TR cells), which act to prevent activation of autoreactive T cells. Here we describe a second mechanism of immune induction by TLRs, which is independent of effects on costimulation. Microbial induction of the Toll pathway blocked the suppressive effect of CD4+CD25+ TR cells, allowing activation of pathogen-specific adaptive immune responses. This block of suppressor activity was dependent in part on interleukin-6, which was induced by TLRs upon recognition of microbial products. PMID- 12532025 TI - Carbon nanotube flow sensors. AB - We report that the flow of a liquid on single-walled carbon nanotube bundles induces a voltage in the sample along the direction of the flow. The voltage that was produced fit a logarithmic velocity dependence over nearly six decades of velocity. The magnitude of the voltage depended sensitively on the ionic conductivity and on the polar nature of the liquid. Our measurements suggest that the dominant mechanism responsible for this highly nonlinear response involves a direct forcing of the free charge carriers in the nanotubes by the fluctuating Coulombic field of the liquid flowing past the nanotubes. We propose an explanation based on pulsating asymmetric ratchets. Our work highlights the device potential for nanotubes as sensitive flow sensors and for energy conversion. PMID- 12532026 TI - Deja vu. PMID- 12532027 TI - 2003 -- new year, new problems? New year, old problems? PMID- 12532028 TI - New approaches to a timeless dilemma. PMID- 12532029 TI - The convexity controversy. PMID- 12532030 TI - Aristotelian logic in home care. PMID- 12532031 TI - Beyond safety and the nursing shortage: a threat to the heart of nursing. AB - Patient safety has been the focus of the recent debate regarding the present nursing shortage. Although important, this focus tends to be short sided. Likely, the nursing shortage threatens all aspects of patient care because it threatens the very essence of healthcare. This article briefly describes the current status of the nursing shortage and argues that this shortage is unique in a number of ways and may be leading to what futurists describe as a long-term crisis. A first person description of nursing is included to further illustrate the threat of the shortage on the heart of patient care. Ethical questions are raised and ideas are presented, including an argument for justifying health care as a special social good. PMID- 12532032 TI - A proposed method for quantifying low-air-loss mattress performance by moisture transport. AB - Because they are believed to control the microclimate of the skin by removing or reducing perspiration accumulation and providing localized cooling, low-air-loss mattress systems are used for the treatment and prevention of pressure ulcers. However, no clear, universally agreed upon definition exists for their design, and reproducible standards on which to base their performance or assess their anticipated clinical effect are lacking. A clinically relevant, reproducible, mechanistic, controlled laboratory test methodology was developed to assess and compare the moisture transport properties of a variety of low-air-loss products by measuring and balancing the complete moisture transport into and out of low air-loss mattress systems. Using a controlled and defined operating environment, the low-air-loss system is moisture- and weight-loaded using a patient skin moisture analog. Moisture and air transport properties into and out of the environment are measured. Total moisture balance, comparing total moisture change of the analog against the time-based moisture transport data, validates the results. Using mattresses from various manufacturers, time-based data using the study method showed important differences in low-air-loss characteristics related to mattress system design, performance, and function. The observed time-averaged moisture transport performance values indicate that several systems meet an acceptable minimum level of performance, but performance levels between different low-air-loss mattress systems vary markedly. PMID- 12532033 TI - The 10(5) bacterial growth guideline: reassessing its clinical relevance in wound healing. AB - The microbiology of wounds is a key determinant in healing and clinicians generally accept that a level of microbial (ie, bacterial) growth greater than 100,000 viable organisms per gram of tissue can be used to diagnose infection. Although other factors that predispose a wound to infection are widely recognized, today's wound care practitioners are influenced primarily by the 10 5 guideline, with treatment being based on the microbial count in deep or superficial tissue. However, to appropriately manage microbially challenged wounds (eg, heavily colonized and clinically infected), a more balanced awareness of the broader issues relating to micro-organisms and wounds is needed. The types of micro-organisms, their interactions with each other and with the wound environment, the local conditions, and host resistance are all key factors that collectively influence healing. From a microbiological perspective, successful wound healing is dependent on maintaining a host-manageable bioburden. If local conditions favor microbial growth, a wound may fail to heal and become infected, requiring topical antiseptics or antibiotics to supplement the host inflammatory response and restore balance in favor of the host. This paper provides a critical examination of the 10 5 guideline to enhance clinician understanding and utilization of a commonly applied diagnostic consideration. PMID- 12532034 TI - Validation of Wagner's classification: a literature review. AB - To ensure high quality care, practitioners should base their practice on sound clinical evidence. Relying on quantitative measurements when evaluating this evidence demands measurement accuracy, reliability, and validity. Current diabetic foot ulcer classification systems include: Meggitt's, Wagner's, Knighton's, Pecoraro's, University of Texas San Antonio Diabetic Wound Classification, and Size (Area and Depth), Sepsis, Arteriopathy, and Denervation. Of these, the Wagner's classification system is the most widely used to describe the natural history of the dysvascular foot, even though evidence of its validity and reliability are lacking. A review of clinimetric properties and existing validation literature of the other diabetic classification systems suggest that the University of Texas San Antonio Diabetic Wound Classification and S(AD) SAD have the potential to fill the current void, once additional studies have been conducted. PMID- 12532035 TI - No-rinse, one-step bed bath: the effects on the occurrence of skin tears in a long-term care setting. AB - As the population ages, the occurrence of skin tears becomes a common concern for those providing care. Skin tears are painful and increase the cost of caring for residents in long-term care facilities. Records of 29 bed-bound residents at a 72 bed, long-term care facility were reviewed to ascertain the effects of changing from using soap and water to a no-rinse formula for bathing on the occurrence of skin tears. The effect of the bathing change on nursing practice, patient care outcomes, and cost of patient care also were examined. When the new bath procedure was used, the number of skin tears decreased from 13 in the first month to one in the fourth month, affecting both caregiver time and cost. The observed reduction in the occurrence of skin tears was estimated to result in an annual decrease in cost $2,446. PMID- 12532036 TI - William Bateson: a biologist ahead of his time. AB - William Bateson coined the term genetics and, more than anybody else, championed the principles of heredity discovered by Gregor Mendel. Nevertheless, his reputation is soured by the positions he took about the discontinuities in inheritance that might precede formation of a new species and by his reluctance to accept, in its full-blooded form, the view of chromosomes as the controllers of individual development. Growing evidence suggests that both of these positions have been vindicated. New species are now thought to arise as the result of genetic interactions, chromosomal rearrangements, or both, that render hybrids less viable or sterile. Chromosomes are the sites of genes but genes move between chromosomes much more readily than had been previously believed and chromosomes are not causal in individual development. Development, like speciation, requires an understanding of the interactions between genes and the interplay between the individual and its environment. PMID- 12532037 TI - Screening for homozygosity by descent in families with autosomal recessive retinitis pigmentosa. AB - Retinitis pigmentosa (RP) is a genetically heterogeneous disease and an important cause of blindness in the state of Andhra Pradesh in India. In an attempt to identify the disease locus in families with the recessive form of the disease, we used the approach of screening for homozygosity by descent in offspring of consanguineous and nonconsanguineous families with RP. Microsatellite markers closely flanking 21 known candidate genes for RP were genotyped in parents and affected offspring to determine whether there was homozygosity at these loci that was shared by affected individuals of a family. This screening approach may be a rapid preliminary method to test known loci for possible cosegregation with disease. PMID- 12532038 TI - Candidate gene polymorphisms among North Indians and their association with schizophrenia in a case-control study. AB - Knowledge of candidate gene polymorphisms in a population is useful for a variety of gene-disease association studies, particularly for some complex traits. A number of candidate genes, a majority of them from the monoaminergic pathway in the brain, have been very popular in association studies with schizophrenia, a neuropsychiatric disorder. In this study diallelic/multiallelic polymorphisms in some dopaminergic, serotonergic and membrane-phospholipid-related genes have been evaluated in a control population recruited from North India. Association, if any, of these allelic variants with schizophrenia has been tested using a case control approach. The case data have been taken from our published family-based association studies in schizophrenia. Of the eight genes tested in this study, association with schizophrenia was observed for only two gene polymorphisms, one in the promoter region of the serotonin 2A receptor gene and the other in the tryptophan hydroxylase gene. One new allele for the dopamine transporter gene (with eight repeats, 570-bp size), not reported in any population so far, has been identified in one individual in our sample. The data generated in this study, besides providing a normative background for various disease association studies, are a significant contribution to the population-specific genome database, a currently growing requirement. PMID- 12532039 TI - Mobile genetic elements in protozoan parasites. AB - Mobile genetic elements, by virtue of their ability to move to new chromosomal locations, are considered important in shaping the evolutionary course of the genome. They are widespread in the biological kingdom. Among the protozoan parasites several types of transposable elements are encountered. The largest variety is seen in the trypanosomatids-Trypanosoma brucei, Trypanosoma cruzi and Crithidia fasciculata. They contain elements that insert site-specifically in the spliced-leader RNA genes, and others that are dispersed in a variety of genomic locations. Giardia lamblia contains three families of transposable elements. Two of these are subtleomeric in location while one is chromosome-internal. Entamoeba histolytica has an abundant retrotransposon dispersed in the genome. Nucleotide sequence analysis of all the elements shows that they are all retrotransposons, and, with the exception of one class of elements in T. cruzi, all of them are non long-terminal-repeat retrotransposons. Although most copies have accumulated mutations, they can potentially encode reverse transcriptase, endonuclease and nucleic-acid-binding activities. Functionally and phylogenetically they do not belong to a single lineage, showing that retrotransposons were acquired early in the evolution of protozoan parasites. Many of the potentially autonomous elements that encode their own transposition functions have nonautonomous counterparts that probably utilize the functions in trans. In this respect these elements are similar to the mammalian LINEs and SINEs (long and short interspersed DNA elements), showing a common theme in the evolution of retrotransposons. So far there is no report of a DNA transposon in any protozoan parasite. The genome projects that are under way for most of these organisms will help understand the evolution and possible function of these genetic elements. PMID- 12532040 TI - Patch testing in discoid eczema. AB - Fifty patients of both sexes with clinically suspected discoid eczema were patch tested with the Indian Standard Patch Test Battery approved by the Contact and Occupational Dermatosis Forum of India (CODFI). Most of these patients were young adults between 20 and 39 years old, with a mean age of 36 years. The mean duration of symptoms was 21/2 years. The occupational profiles of the patients included farmers (24%), housewives (18%), students (14%), housewives engaged in farming (12%), shopkeepers (10%), and laborers (8%). The miscellaneous group, including technicians, drivers, cable operators, office workers, and computer operators, accounted for the remaining 14%. The common clinical patterns of presentation of discoid eczema included hands and feet (44%), hands and forearms' (30%), legs and feet (12%) and trunk and limbs (12%). Patch test analysis of these patients revealed that, out of the 50 subjects tested, 28 (56%) reacted to one or more allergens. Potassium dichromate was the most common allergen (20%), followed by nickel (16%), cobalt chloride, and fragrance (12% each) in decreasing order of frequency. Hence, it may be concluded that patch testing should be considered for all patients with severe or persistent discoid eczema because allergic contact dermatitis may be relatively common in such patients and the avoidance of offending allergens may be of substantial benefit to the patients. PMID- 12532041 TI - A keratin 9 Gene mutation (Asn160Ser) in a Japanese patient with epidermolytic palmoplantar keratoderma. AB - We described a 5-year-old Japanese girl with epidermolytic palmoplantar keratoderma and examined her for a keratin 9 gene mutation. Physical examination disclosed diffuse yellowish hyperkeratosis with an erythematous border limited strictly to the palms and soles. Histological examination revealed hyperkeratosis with vacuolar degeneration in the spinous and granular layers of the epidermis. Sequence analysis demonstrated an A to G transition at the middle position of codon 160 in the 1A domain of the keratin 9 gene. The amino acid at codon 160 was deduced to have changed from asparagine (Asn) to serine (Ser). This is the first case with an Asn160Ser mutation in a Japanese. The substitution of Ser for Asn at codon 160 of the keratin 9 gene is assumed to be fatal for keratin filament assembly regardless of race or ethnicity. PMID- 12532042 TI - Psoriasis vulgaris and Poland's syndrome in the same patient. AB - Most common anomalies associated with Poland's syndrome are in the chest wall. This type of anomaly causes important psychological distress, especially in adolescent girls. A 19-year-old female with Poland's syndrome having psoriatic plaques on her trunk, knees, and elbows is reported in this paper. Such an association of Poland's syndrome with psoriasis has not been described previously. PMID- 12532043 TI - Antiphospholipid antibody syndrome complicated by Grave's disease. AB - The report describes a woman with primary antiphospholipid antibody syndrome complicated with Grave's disease. Developing symptoms included a small cutaneous nodule on her finger and subsequently ecchymotic purpura on the cheeks, ears, buttocks and lower legs. Histological examinations showed thrombosed vessels in the dermis without or with hemorrhage, respectively. Laboratory investigation revealed positive lupus anticoagulant and immunogenic hyperthyroidism due to Grave's disease. There is a close relationship between the cutaneous manifestation of antiphospholipid antibody syndrome and the activities of Grave's disease and a possible link of antiphospholipid antibody syndrome with Grave's disease was suggested both by the etiology of the disease as well as the disease activity. PMID- 12532044 TI - Childhood bullous pemphigoid following hepatitis B immunization. AB - Bullous pemphigoid (BP) is an acquired autoimmune bullous disorder which predominantly affects the elderly. It is very rare in children. There are approximately 50 reported cases of childhood BP. Although the cause of childhood BP is unknown, drug intake and vaccination have been incriminated in some cases. A total of 13 patients with BP (10 adults and 3 infants) have been described to be related to various vaccines and tetanus toxoid booster. However, no case related to hepatitis B vaccination has previously been reported. Our case of childhood BP developed one week after hepatitis B immunization in a Turkish caucasian child. This case suggests that the hepatitis B surface antigen can function as the triggering factor for BP by inducing a nonspecific immune reactivation which unmasks subclinical BP or by stimulating a specific antibody production that may cross-react with BP antigens. PMID- 12532045 TI - A case of polypoid dermatofibroma. AB - Dermatofibroma is a common benign cutaneous tumor that usually appears as a slowly growing firm nodule. Polypoid nodular dermatofibroma is a variant type that is rarely encountered. We reported a case of polypoid dermatofibroma with a review of the previously reported cases. Polypoid dermatofibroma tends to arise on the leg, especially below the knee. Its size is often larger than that of common dermatofibroma. It is speculated that both the underlying firm tissue and long-term development may lead the tumor to form a polypoid appearance. PMID- 12532046 TI - Carney complex: report of a Japanese case associated with cutaneous superficial angiomyxomas, labial lentigines, and a pituitary adenoma. AB - We report the case of a 12-year-old female patient who manifested multiple cutaneous angiomyxomas and labial pigmented lesions. Although the familial history was not confirmed in the present case, autosomal dominant inheritance has been reported to be involved in the pathogenesis of this condition. In addition to the cutaneous complications, magnetic resonance (MR) images revealed the presence of a pituitary adenoma, which provoked an elevation of serum growth hormone (GH) level. On the other hand, no significant symptoms such as cardiac myxoma, myxoid fibroadenoma of the breast, or adrenocortical complaints suggesting Cushing syndrome, were detected. In the Japanese literature, only a few cases of this disorder have been described in the form of brief reports. There have been only a few similar cases described in the dermatological field, except for one report diagnosed as Carney complex in 1990. Therefore, the present case seems to be the first Japanese case of typical Carney complex manifesting major clinical complications, including angiomyxomas, lentigines, and a pituitary adenoma, which induced endocrine overactivity. PMID- 12532047 TI - A case of primary cutaneous histoplasmosis in a patient with diabetes and multi infarct dementia. AB - We report a case of primary cutaneous histoplasmosis in a fifty-year-old African American woman with diabetes and multi-infarct dementia. The patient developed fever and crusted, nodulo-ulcerative lesions of the skin after accidental superficial trauma to the forehead. The biopsy revealed suppurative granulomatous inflammation with intracellular and extracellular yeast-like cells with associated clear halo measuring 3-4 mm in size. Systemic involvement was not found. The lesions cleared after treatment with itraconazole 200 mg twice a day for 3 weeks. The medication was continued for a total period of 3 months, with no signs of recurrence after one-year of follow-up. PMID- 12532048 TI - Cutaneous nocardiosis developing around gravel inserted during a traffic injury. AB - A 65-year-old Japanese woman with nocardial mycetoma which developed five years after a compound bone fracture of the left foot due to a traffic accident was reported. During this accident, two small pieces of gravel had entered her foot. Nocardia asteroides, which had been attached to the gravel, was probably inserted 5 years previously during the trauma. The lesion did not respond to oral minocycline and trimethoprim-sulfamethoxazole and was finally removed surgically; she has been free from recurrences for one year, of follow-up. PMID- 12532049 TI - Efficacy of oral minocycline and hyperthermic treatment in a case of atypical mycobacterial skin infection by Mycobacterium marinum. AB - We report a case of atypical mycobacterial dermal infection caused by M. marinum, which was effectively treated with oral administration of minocycline and local hyperthermic treatment using chemical pocket warmers. A daily oral dose of 200 mg of minocycline was given, and local hyperthermic treatment was applied every evening for 5-6 hours with a disposable chemical pocket warmer. After 2.5 months of therapy, the lesion healed completely with scar formation. At 24 months after the completion of treatments, there is no sign of recurrence. PMID- 12532050 TI - Psoriasis arthropathy and lymphedema. PMID- 12532051 TI - Minor Poland's syndrome mimicking localized lipoatrophy. PMID- 12532052 TI - Matting of scalp hair. PMID- 12532053 TI - A case of facial pomade crust. PMID- 12532054 TI - Manpower and the changing economy. PMID- 12532055 TI - Comparison of in vivo and in vitro shear bond strength. AB - Much of the research into composite adhesives suggests that these materials will deteriorate in the oral environment, but most of these claims are made on the basis of extrapolation from in vitro experiments; relatively little in vivo research has been conducted into the mechanical properties of composite adhesives. For this study, we recruited 20 volunteers to wear removable appliances containing orthodontic brackets bonded to enamel slabs for 12 weeks. Each appliance carried 3 brackets bonded with Transbond (3M Unitek UK; Bradford, United Kingdom) and 3 brackets bonded with Heliosit (Ivoclar-Vivadent; Leicester, United Kingdom). The bond strengths were tested at intervals of 4, 8, and 12 weeks. Control specimens were stored in sterile water at 37 degrees C and were debonded at the same time intervals. Transbond specimens debonded after 4 weeks in vivo had significantly (P <.05) lower bond strengths (9.78 megapascals [MPa]) than did the controls (14.34 MPa). In vivo, specimens bonded with Heliosit had significantly (P <.05) lower bond strengths after 4 weeks (8.16 MPa vs 10.96 MPa) and 8 weeks (9.96 MPa vs 13.61 MPa) than did the controls. These results indicate differences between bond strength testing in vitro and in vivo. Further research is required into the effects of the oral environment on bond strength. PMID- 12532056 TI - In vivo inhibition of demineralization around orthodontic brackets. AB - Demineralization around orthodontic appliances is a problem. Suboptimal oral hygiene, long intervals between appointments, and potentially poor patient cooperation with using fluoride dentifrices and mouth rinses necessitate a compliance-free means of preventing tooth decay. The hypothesis of this study was that fluoride released by glass ionomer cement inhibits the formation of carious lesions around orthodontic brackets in vivo. Brackets were bonded on 2 first premolars in 21 randomized, consecutively selected patients 11 to 18 years old. Eleven test-group subjects were bonded with fluoride-releasing glass ionomer cement, and 10 control subjects were bonded with composite resin (no fluoride). The teeth were extracted after 4 weeks, sectioned, and evaluated quantitatively by cross-sectional microhardness testing. Fluoride levels in patient saliva were measured by the Taves diffusion method in samples taken at days 0 (baseline), 1, 2, 3, 7, 14, 21, and 28 to determine whether fluoride from the glass ionomer cement influenced the overall intraoral fluoride levels. The results demonstrated significantly more demineralization around the brackets of the control patients (P <.01, Wilcoxon signed rank test). For whole-mouth salivary fluoride levels, no significant overall difference between the groups (P >.05) and no noticeable trend within groups (P >.05) were found. These results indicate that using fluoride-releasing glass ionomer cement for bonding orthodontic brackets successfully inhibited caries in vivo. This cariostatic effect was localized to the area around the brackets and was statistically significant after 4 weeks. PMID- 12532057 TI - In vitro comparison of orthodontic band cements. AB - The aim of the study was to compare the mean retentive strength of microetched orthodontic bands cemented to extracted human third molars with a modified composite, a resin-modified glass ionomer cement, and a conventional glass ionomer cement. The mode of band failure and amount of cement remaining on the tooth at deband were also assessed. Finally, survival time of bands with each cement was assessed with simulated mechanical stress in a ball mill. Ninety banded specimens were used to assess retentive strength, and another 30 banded specimens were used to assess survival time. The mean retentive strength of the modified composite (0.415 MPa) was significantly less than that of either the resin-modified (1.715 MPa) or the conventional glass ionomer cement (1.454 MPa; P <.001). Specimens failed predominantly at the cement-enamel interface. The amount of cement remaining on the tooth at deband differed significantly between bands cemented with the resin-modified cement and those cemented with the conventional glass ionomer cement (P <.05). Mean survival time of bands cemented with the resin-modified glass ionomer cement (14.3 hours) was significantly longer (P <.01) than for bands cemented with the conventional glass ionomer cement (9.9 hours) but did not differ significantly from that of bands cemented with the modified composite (11.1 hours; P >.05). Orthodontic bands cemented with the modified composite appear to have a significantly lower mean retentive strength than bands cemented with resin-modified or conventional glass ionomer cement, but mean survival time did not differ significantly for bands cemented with modified composite or resin-modified glass ionomer. PMID- 12532058 TI - Effect of thermocycling on the shear bond strength of a cyanoacrylate orthodontic adhesive. AB - The purpose of this study was to evaluate the effects of thermocycling on the shear bond strength of a cyanoacrylate adhesive system, specifically 24 hours after bonding when the adhesive has achieved most of its bond strength and after thermocycling. Forty freshly extracted human molars were collected and stored in a solution of 0.1% (weight/volume) thymol. The teeth were cleaned, polished, and randomly separated into 2 groups: group I, cyanoacrylate adhesive debonded after 24 hours immersion in deionized water at 37 degrees C; and group II, cyanoacrylate adhesive debonded after thermocycling at 5 degrees C and 55 degrees C. The results of the t test comparing the 2 groups (t = 6.84) indicated significant differences between them (P =.0001). The cyanoacrylate adhesive at 24 hours had significantly greater shear bond strength (macro x = 7.1 +/- 3.3 MPa) than after thermocycling 500 times between 5 degrees C and 55 degrees C (macro x = 1.5 +/- 1.4 MPa). The findings indicated that the cyanoacrylate adhesive tested has clinically adequate shear bond strength at 24 hours after initial bonding but loses about 80% of its strength after thermocycling. The clinician should consider all properties of the adhesive, including no need for a curing light, working time of 5 seconds before the adhesive starts to set, and the significant decrease in bond strength after thermocycling. PMID- 12532059 TI - Arch width after extraction and nonextraction treatment. AB - An undocumented criticism of extraction treatment is that it results in narrower dental arches when compared with nonextraction therapy. Anterior and posterior arch widths of the maxillary and mandibular arches of 25 patients treated by 4 first-premolar extraction and 25 patients treated without extractions were measured on posttreatment study models and compared statistically to determine whether the dental arches were narrower after extraction treatment, to test the accuracy of this view. Measurements were made in the canine and the molar regions from the most labial aspect of the buccal surfaces of the canines and the molars. In both groups, anterior and posterior arch widths were the same except for the mandibular intercanine dimension, which was 0.94 mm larger (P <.01) in the extraction group. This indicates that extraction treatment does not result in narrower dental arches than nonextraction treatment. PMID- 12532060 TI - Tongue pressure on loop of transpalatal arch during deglutition. AB - The purpose of this study was to measure tongue pressure exerted on the loop of the transpalatal arch (TPA) during deglutition and to consider the influence of the distance of the loop of the TPA from the palatal mucosa and the anteroposterior position of the loop. Tongue pressures of 4 subjects with normal occlusion were measured with subminiature pressure sensors fixed on the TPA. The distances from the palatal mucosa to the surface of the pressure sensor were set at 2, 4, and 6 mm. The loop of the TPA was placed at the level of the middle of the maxillary second premolars (P), first molars (M1), or second molars (M2). Nine types of TPA devices were measured for each subject. The maximum recorded tongue pressure was taken from each act of deglutition. The minimum pressure value was exerted at position P when the distance from the palatal mucosa to the surface of the pressure sensor was 2 mm. The maximum value was obtained at position M2 and a distance of 6 mm from the palatal mucosa. When distances of 2, 4, and 6 mm were compared, significant differences between 2 and 4 mm, and between 2 and 6 mm were found. Significant differences were observed in comparisons between the positions P and M1, M1 and M2, and P and M2. PMID- 12532061 TI - Familial correlations and heritability of maxillary midline diastema. AB - The purpose of the study was to estimate familial correlations and heritability to evaluate familial aggregation patterns of maxillary midline diastemas. The sample consisted of 30 extended families: 15 black, 14 white, and 1 mixed race. A single ascertainment scheme was adopted to collect the sample. Family data were collected with a 7-question survey. In all, the sample of 430 subjects consisted of 220 females, 210 males, 99 nuclear families, 534 sibling pairs, 422 avuncular pairs, 318 grandparent pairs, and 27 cousin pairs. Families were stratified by race to avoid any bias. The mixed-race family was excluded from the analysis. Data were analyzed using the program REGC in the Statistical Analysis for Genetic Epidemiology (S.A.G.E., Case Western Reserve University, Cleveland, Ohio) software. Heritability was found to be 0.32 +/- 0.14 in the white sample and 0.04 +/- 0.16 in the black sample. The preliminary results suggest a possible genetic basis for maxillary midline diastema and a greater role of environmental factors in the black sample than in the white sample. PMID- 12532062 TI - Functional appliance therapy accelerates and enhances condylar growth. AB - The present study was designed to quantitatively assess the temporal pattern of expression of Sox 9, the regulator of chondrocyte differentiation and type II collagen, the major component of the cartilage matrix during forward mandibular positioning, and compare it with the expression during natural growth. Female Sprague-Dawley rats, 5 weeks old, were used. Results showed that the expression of Sox 9 and type II collagen are accelerated and enhanced when the mandible is positioned forward. Furthermore, we monitored the amount of new bone formation during mandibular advancement and after the removal of bite-jumping appliances. A substantial increase was observed in the amount of newly formed bone when the mandible was positioned forward. No significant difference in new bone formation could be found after the appliance was removed when compared with natural growth. Thus, functional appliance therapy accelerates and enhances condylar growth by accelerating the differentiation of mesenchymal cells into chondrocytes, leading to an earlier formation and increase in amount of cartilage matrix. This enhancement of growth did not result in a subsequent pattern of subnormal growth for most of the growth period; this indicates that functional appliance therapy can truly enhance condylar growth. PMID- 12532063 TI - Replicating mesenchymal cells in the condyle and the glenoid fossa during mandibular forward positioning. AB - The purpose of this study was to identify and quantify the temporal sequence of replicating mesenchymal cells during natural growth and mandibular advancement in the condyle and the glenoid fossa. One hundred fifty 35-day-old female Sprague Dawley rats were randomly divided into 10 experimental groups (10 rats each) and 10 control groups (5 rats each). The experimental groups were fitted with appliances that positioned the mandible forward. One hour before the rats were killed, bromodeoxyuridine (BrdU) was intravenously injected into them. Sections were cut and stained with anti-BrdU antibody to evaluate the number of replicating mesenchymal cells. Cellular uptake of BrdU was quantified with the Leica Qwin (Leica Microsystem Imaging Solutions, Cambridge, United Kingdom) system. The results showed that the numbers of replicating mesenchymal cells during natural growth were highest in the posterior region of the condyle and the anterior region of the glenoid fossa. In the experimental groups, the posterior region had the highest number of replicating cells for both the condyle and the glenoid fossa, with the condyle having 2 to 3 times more replicating cells than the glenoid fossa. The number of replicating mesenchymal cells, which is genetically controlled, influences the growth potential of the condyle and the glenoid fossa. Mandibular protrusion leads to an increase in the number of replicating cells in the temporomandibular joint. Individual variations in the response to growth modification therapy could be a result of the close correlation between mesenchymal cell numbers and growth. PMID- 12532064 TI - Effects of upper lip closing force on craniofacial structures. AB - Force generated by the perioral musculature is known to be a potent factor that can affect tooth position and malocclusion. To evaluate the influence of the force of the orbicularis oris muscle on incisor position, we determined the maximum and average strengths of the upper lip of male Class I malocclusion patients with a Y-meter, which was devised to measure the vertical closing force of the lip with a load cell. The skeletal structures and incisor angulation were recorded by lateral cephalograms. Correlation and stepwise regression analyses were performed to determine whether a relationship existed between lip force levels and craniofacial morphology. The average value of the upper lip closing force (AVE) was 7.16 N. Our results showed that the upper incisor angulation was related to the perioral muscle force. The maxillary incisor to Frankfort horizontal angle showed r values of -0.681 (at AVE) and -0.652 (at maximum lip closing force) as determined by correlation analysis. Stepwise regression analysis also showed that the maxillary incisor to Frankfort horizontal angle was more related to the upper lip closing force. Disuse atrophy of the orbicularis oris seems to be an important factor in the development of malocclusion. PMID- 12532065 TI - Three-dimensional relationship between the critical contact angle and the torque angle. AB - The purpose of this study was to analyze the relationship between the critical contact angle and the torque angle in an orthodontic bracket and archwire assembly in 3 dimensions. Three-dimensional mathematical models were created with geometric bracket-archwire parameters that included 2 slot sizes, 3 bracket widths, and 3 to 4 wire sizes. From this, 3-dimensional mathematical equations (3DMEs) for the critical contact angle and the maximum torque that result in critical contact angles of 0 were derived and calculated. To evaluate the effects of archwire-bracket parameters on critical contact angles, analysis of variance (ANOVA) was performed at the significance level of P < or = .05. For all bracket archwire combinations, the critical contact angle decreased as bracket width, torque angle, and wire size increased. Therefore, all bracket-archwire parameters except slot height had an effect on the critical contact angle. Results of the critical contact angle produced from our 3DMEs were the same as those produced by 3D computer-aided design (SolidWorks Corp, Concord, Mass), thus confirming the validity of our derived equations. In addition, the effect of a beveled edge was investigated in some archwires. Furthermore, torsional play angles were calculated and found to be similar to those in previous reports. The results of this study provide theoretic and experimental bases for clinical orthodontic practice and indicate that torque angles should be included in the evaluation of the critical contact angle. PMID- 12532066 TI - Orthodontic treatment considerations in patients with diabetes mellitus. AB - Diabetes mellitus (DM) is diagnosed in 3% to 4% of the population. This metabolic disorder is characterized by hyperglycemia caused by the body's deficient management of insulin. Two main types of DM exist: type 1 is a total deficiency in insulin secretion, and type 2 is a combination of resistance to insulin action and inadequate compensatory insulin secretion. Diagnosis and monitoring of DM is based on blood glucose concentration or glycosylated hemoglobin concentration. The dental practitioner should be aware of the oral manifestations of DM in order to spot initial symptoms of the disease. When treating DM patients, the practitioner must understand the consequences of the disease in relation to dental treatment. In this review, we examine the implications of DM when orthodontic treatment is planned. PMID- 12532067 TI - Laser debonding of ceramic brackets: a comprehensive review. AB - Since the invention of the ruby laser in the early 1960s, tremendous advances have been made in optic laser technology. Orthodontists have found various uses for lasers, including the debonding of ceramic brackets. Laser energy degrades the adhesive resin used to bond brackets. Consequently, lower forces can be used than when mechanical debonding is performed, reducing the risk of enamel damage. However, the heat produced by some lasers can damage the tooth pulp. Selecting the appropriate laser, resin, and bracket combination can minimize risks and make debonding more efficient. The purpose of this article is to give the clinician an up-to-date, comprehensive literature review about the clinical characteristics of debonding ceramic brackets with lasers. PMID- 12532068 TI - Segmental odontomaxillary dysplasia: an unusual orthodontic challenge. AB - Segmental odontomaxillary dysplasia has only recently been delineated from other osseo-odontogenic dysplasias. Yet, this seldom-reported developmental abnormality might be diagnosed by a unique combination of clinical, radiographic, and histologic findings. Unilateral maxillary hyperplasia combined with displaced molars and unusual bone trabeculation is quite obvious at an early age. Recognition and subsequent reporting by dental specialists is encouraged to more clearly define the natural progression and the appropriate treatment of this abnormality. PMID- 12532069 TI - Unerupted maxillary central and lateral incisors and canine with crossbite and asymmetry. AB - This case report describes the treatment of a patient with unerupted maxillary right central incisor, lateral incisor, and canine in a Class III malocclusion with asymmetry of the lower face. Sequential traction of the 3 unerupted teeth with a light force system was used in the treatment. The results remained stable during the 4-year retention period. PMID- 12532070 TI - Litigation, legislation, and ethics. Subjective expert opinion evidence. PMID- 12532071 TI - Science versus art in case management. PMID- 12532072 TI - Reducing professional stressors: making work work for you! PMID- 12532073 TI - Case management and patient safety. PMID- 12532074 TI - Supreme court agrees to define employer. PMID- 12532075 TI - Faith and healing. PMID- 12532077 TI - Staying sharp. Keeping up with advances in medical technology. PMID- 12532078 TI - Case management best practices that pave the way for real world success. PMID- 12532079 TI - Tub benches for people with spinal cord injury: luxury or necessity? PMID- 12532080 TI - Pulmonary hypertension. Real life stories. PMID- 12532081 TI - Beware costly tax penalties when using independent contractors. PMID- 12532082 TI - Introduction: the anti-inflammatory role of cysteinyl leukotriene receptor antagonists in asthma. PMID- 12532083 TI - Inflammatory cells in asthma: mechanisms and implications for therapy. AB - Recent clinical studies have brought asthma's complex inflammatory processes into clearer focus, and understanding them can help to delineate therapeutic implications. Asthma is a chronic airway inflammatory disease characterized by the infiltration of airway T cells, CD(+) (T helper) cells, mast cells, basophils, macrophages, and eosinophils. The cysteinyl leukotrienes also are important mediators in asthma and modulators of cytokine function, and they have been implicated in the pathophysiology of asthma through multiple mechanisms. Although the role of eosinophils in asthma and their contribution to bronchial hyperresponsiveness are still debated, it is widely accepted that their numbers and activation status are increased. Eosinophils may be targets for various pharmacologic activities of leukotriene receptor antagonists through their ability to downregulate a number of events that may be key to the effector function of these cells. PMID- 12532084 TI - Roles of cysteinyl leukotrienes in airway inflammation, smooth muscle function, and remodeling. AB - A new paradigm for asthma pathogenesis is presented in which exaggerated inflammation and remodeling in the airways are a consequence of abnormal injury and repair responses arising from a subject's susceptibility to components of the inhaled environment. An epithelial-mesenchymal trophic unit becomes activated to drive pathologic remodeling and smooth muscle proliferation through complex cytokine interactions. Histamine, prostanoids, and cysteinyl leukotrienes (CysLTs) are potent contractile agonists of airway smooth muscle (ASM). The CysLTs appear to play a central role in regulating human ASM motor tone and phenotypic alterations, manifested as hypertrophy and hyperplasia in chronic severe asthma. The CysLTs augment growth factor-induced ASM mitogenesis through activation of CysLT receptors. Although they mediate their contractile effects by increasing phosphoinositide turnover and inducing increased cytosolic calcium, new data suggest that part of the contractile effect may be independent of calcium mobilization. Prostaglandin E(2), the predominant eicosanoid product of the airway epithelium, is a potent inhibitor of mitogenesis, collagen synthesis, and mesenchymal cell chemotaxis and therefore can suppress inflammation and fibroblast activation. The capacity of the epithelium for CysLT synthesis is inversely related to its ability to make PGE(2). The ASM is capable of expressing both leukotriene-synthesizing enzymes and CysLT receptors, and cytokines upregulate the receptor expression. This may be an explanation for the CysLTs promoting airway hyperresponsiveness in asthma. The CysLTs play an important role in the airway remodeling seen in persistent asthma that includes increases of airway goblet cells, mucus, blood vessels, smooth muscle, myofibroblasts, and airway fibrosis. Evidence from a mouse model of asthma demonstrated that CysLT(1) receptor antagonists inhibit the airway remodeling processes, including eosinophil trafficking to the lungs, eosinophil degranulation, T(H)2 cytokine release, mucus gland hyperplasia, mucus hypersecretion, smooth muscle cell hyperplasia, collagen deposition, and lung fibrosis. PMID- 12532085 TI - Cysteinyl leukotriene interactions with other mediators and with glucocorticosteroids during airway inflammation. AB - Unexpected aspects of the antiasthmatic efficacy of leukotriene modifiers and glucocorticosteroids have been observed. For both classes, the observed effects may be partially explainable on the basis of underrecognized interactions involving leukotrienes. This review examines the interactions between leukotrienes and other mediators of asthma. It details the effects of glucocorticosteroids on leukotriene synthesis and on leukocyte populations in asthmatic airways. Unexpected controller effects of the leukotriene modifiers may reflect the fact that leukotrienes and other mediators of asthma, such as T(H)2 cytokines, positively influence each other's generation. The ability of the leukotriene modifiers to disrupt such extensive interactions means that other relevant mediators are targeted indirectly by leukotriene blockade. Among asthma therapies, the glucocorticosteroids have numerous anti-inflammatory activities, but their effects may be unpredictable. Many processes involved in inflammation appear to escape modulation by glucocorticosteroids, including leukotriene synthesis, and leukotriene generation is among them. Understanding whether glucocorticosteroids reduce cysteinyl leukotriene levels in the airway is important in determining the clinical value of combining glucocorticosteroid therapy with leukotriene modifier therapy. PMID- 12532086 TI - Effects of cysteinyl leukotrienes and leukotriene receptor antagonists on markers of inflammation. AB - The understanding that asthma pathophysiology includes an inflammatory component has spurred the more aggressive use of anti-inflammatory therapies and created a need for effective tools to measure inflammation. Biomarkers of airway inflammation proposed are obtained by methods that are direct but highly invasive (bronchial biopsy, bronchoalveolar lavage), moderately direct, and less invasive (indirect sputum, exhaled air, breath condensate) or indirect and least invasive (blood, urine). Several studies described in this review have implicated the cysteinyl leukotrienes (CysLTs) as inflammatory mediators in a wide range of diseases, implying that their biological activities reach far beyond acute bronchoconstriction, the activity traditionally ascribed to them. The validity of examining sputum for "biomarkers" has improved the understanding of asthma pathophysiology, optimization of asthma treatment and management, and investigation of the relation between CysLTs and airway inflammation in asthma. Nitric oxide is also a surrogate marker of asthma and reflects airway inflammation. The anti-inflammatory effects of the leukotriene receptor antagonists and the markers of their activity continue to grow. PMID- 12532087 TI - Leukotriene receptor antagonists in asthma therapy. AB - In persons with asthma, the cysteinyl leukotrienes possess multiple inflammatory properties in vitro and have long been considered to be a potentially important mediator of asthma and an attractive target for therapeutic intervention. Controlled clinical trials have documented the efficacy of leukotriene receptor antagonists in asthma treatment, but reservations about their use for asthma therapy center on two main issues: the heterogeneity of patient responses and their reduced potency relative to other asthma medications. For example, leukotriene receptor antagonists also have been shown to be less efficacious than inhaled corticosteroids for several end points, including symptom relief, reduced markers of inflammation, and improved pulmonary function. This review explores several underappreciated aspects of asthma therapy: heterogeneity of patient responses to medication, the failure of symptoms to correlate with commonly used end points, and the potential of delivery to distal airways for producing important and novel benefits. PMID- 12532089 TI - Update on glucocorticoid action and resistance. AB - Glucocorticoids (GCs) are the most common group of medications used in the treatment of allergic and autoimmune disorders. They produce potent anti inflammatory effects by inducing or repressing the expression of target genes. Although most patients with allergic diseases and autoimmune disorders respond to GC therapy, a small subset of patients demonstrate persistent tissue inflammation despite treatment with high doses of GCs. This condition results from an interaction between susceptibility genes, the host's environment, and immunologic factors. The treatment of these patients requires a systematic approach to rule out underlying conditions that lead to steroid resistance or treatment failure, as well as the use of alternative strategies to inhibit tissue inflammation. PMID- 12532090 TI - Mast cells: beyond IgE. AB - Mast cells, historically known for their involvement in type I hypersensitivity, also serve critical protective and homeostatic functions. They directly recognize the products of bacterial infection through several surface receptor proteins, releasing proteases, cytokines, and eicosanoid mediators that recruit neutrophils, limit the spread of bacterial infection, and facilitate subsequent tissue repair. In vitro studies suggest that the spectrum of microbes capable of initiating mast cell activation is broad and extends to common respiratory viruses, mycoplasma, and even products of tissue injury, such as nucleotides. TH2 polarized inflammation elicits a reactive hyperplasia of mast cells at the involved mucosal surfaces in both mice and human subject. Several recombinant TH2 cytokines (IL-3, IL-4, IL-5, and IL-9) act synergistically with stem cell factor to facilitate proliferation of nontransformed human mast cells in vitro. IL-4 induces the expression of critical inflammation-associated genes by human mast cells, such as those encoding leukotriene C4 synthase, Fc(epsilon)RI, and several cytokines. Consequently, priming with IL-4 not only amplifies classical Fc(epsilon)RI-dependent mast cell activation but also dramatically alters the product profile of mast cells activated by innate signals and by chemical mediators of inflammation. Strikingly, IL-4 induces an activation response by mast cells to cysteinyl leukotrienes, which act through a receptor shared with uridine diphosphate to induce cytokine generation without exocytosis. It Is possible that alterations in mast cell phenotype by the TH2 milieu of allergy permits otherwise trivial infections or homeostatic chemical signals to initiate harmful inflammatory cascades and sustain tissue pathology. Drug development must take these nonclassical mast cell activation pathways into account without compromising the beneficial and protective functions of mast cells. PMID- 12532093 TI - Why must olympic athletes prove that they have asthma to be permitted to take inhaled beta2-agonists? PMID- 12532094 TI - Unraveling the mission of FcepsilonRI on antigen-presenting cells. AB - A decade ago, the discovery of the high-affinity receptor for IgE (FcepsilonRI) on epidermal Langerhans cells documented the end of the dogma that FcepsilonRI is only expressed on effector cells of anaphylaxis. Since then, the functional significance of this receptor on antigen-presenting cells (APCs) has been an area of intense research work. Scientists have focused on a better understanding of the molecular structure, regulation, and role of FcepsilonRI on APCs in the human immune system. Insights into the cellular events linked to the activation of APCs on ligation of FcepsilonRI by IgE and allergens might provide the basis for new aspects in the pathophysiology of allergic diseases and the design of future diagnostic and therapeutic strategies. This review is dedicated to the 10th anniversary of the discovery of FcepsilonRI on APCs and describes the numerous areas of research in this field. PMID- 12532095 TI - Responses to bronchial challenge submitted for approval to use inhaled beta2 agonists before an event at the 2002 Winter Olympics. AB - BACKGROUND: There has been an increase in the number and percentage of athletes competing in Olympic Games notifying use of beta2-agonists, from 1.7% at Los Angeles (1984) to 5.5% at Sydney (2000). For Salt Lake City (2002), the International Olympic Committee requested objective evidence to use beta2 agonists for asthma or exercise-induced asthma (EIA). OBJECTIVE: The objective of this study was to evaluate the evidence submitted for approval to use a beta2 agonist. METHODS: Objective evidence for asthma or EIA included (1) an increase of 12% or more of the predicted FEV1 in response to bronchodilator, (2) a reduction in FEV1 of 10% or greater from baseline in response to exercise or eucapnic voluntary hyperpnea, (3) a PD20 FEV1 to methacholine or histamine at a dose of less than 200 microg (2 mg/mL) or less than 1320 microg (13.2 mg/mL) for those taking inhaled corticosteroids for 3 months. RESULTS: There were 165 applications. Of these, 147 (89%) included evidence of a challenge, bronchodilator response, or both, and 163 test results were submitted. One hundred thirty (5.2%) applications were approved. For those with positive responses, the median value (1) was 16.2% of predicted FEV1 for response to a bronchodilator (n = 13), (2) was a 15.9% decrease in FEV1 for response to a physical challenge (n = 36), and, (3) for PD20 FEV1, was 173 microg for response to a pharmacologic challenge (n = 45). CONCLUSION: The analysis demonstrated that it is feasible to request objective evidence to justify use of beta2-agonists on the medical grounds of asthma or EIA. PMID- 12532096 TI - Mode of delivery and risk of allergic rhinitis and asthma. AB - BACKGROUND: It has been hypothesized that cesarean section might increase the risk of developing allergic disease by depriving the fetus and newborn of exposure to maternal microflora. Furthermore, it has been suggested that complicated modes of delivery might be associated with an increased risk of asthma. OBJECTIVE: The purpose of this investigation was to study whether cesarean section and other complicated modes of delivery are associated with an increased risk of allergic rhinitis or asthma. METHODS: Information on self reported allergic rhinitis, asthma ever, current asthma, and occupation was obtained from 9722 singleton women born in Denmark during the period 1973-1977 who participated in a national cohort study during the period 1997-2001. For these women, information was available on mode of delivery (spontaneous delivery, cesarean section, vacuum extraction, or other complicated mode of delivery, such as rotation/traction or use of forceps), gestational age, birth weight, and length at birth from the Danish Medical Birth Register. Information on parity and maternal age was obtained from the Danish Civil Registration System. RESULTS: The odds ratios (ORs) of allergic rhinitis were 1.16 (95% CI, 0.90-1.49) for cesarean section and 1.06 (95% CI, 0.85-1.32) for other complicated modes of delivery in comparison with spontaneous delivery. The corresponding ORs of asthma ever were 1.33 (95% CI, 1.02-1.74) and 1.18 (95% CI, 0.94-1.49) for cesarean section and other complicated modes of delivery, respectively, and the ORs of current asthma were 1.22 (95% CI, 0.87-1.73) and 1.26 (95% CI, 0.94-1.68), respectively, in comparison with spontaneous delivery. CONCLUSIONS: Our findings do not support the hypothesis that cesarean section or other complicated modes of delivery are associated with the development of allergic rhinitis. However, there might be a positive association with development of asthma--in particular, for cesarean section--that was not explained by gestational age, birth weight, ponderal index, smallness for gestational age, parity, maternal age, or occupation. PMID- 12532097 TI - Steroid-sparing effects of fluticasone propionate 100 microg and salmeterol 50 microg administered twice daily in a single product in patients previously controlled with fluticasone propionate 250 microg administered twice daily. AB - BACKGROUND: Concurrent use of an inhaled corticosteroid (ICS) and an inhaled long acting beta2-agonist provides better overall asthma control than the use of higher doses of ICS alone. OBJECTIVE: The purpose of this investigation was to determine whether fluticasone propionate (FP) combined with salmeterol in the Diskus device can be used to reduce the dose of ICS in patients currently stable on medium-dose ICS while maintaining asthma control. METHODS: This was a randomized, double-blind, parallel-group, 12- to 24-week trial consisting of a 3 part run-in period. The run-in period was designed to first establish FP 250 microg administered twice a day (bid) via Diskus as the minimum effective dose. During run-in period 1, patients received FP 220 microg bid or the equivalent for 10 to 14 days. Controlled patients moved to run-in period 2 (5-28 days), which assessed asthma stability on FP 100 microg bid administered via Diskus. Only patients who became unstable on FP 100 microg bid were eligible to enter run-in period 3 (26-30 days), during which they were placed on FP 250 microg bid and those regaining asthma control were eligible for randomization. The primary efficacy endpoint was the proportion of patients who remained in the study with no evidence of worsening asthma. Secondary efficacy measures included FEV1, morning peak expiratory flow, percent of symptom-free days, and daily albuterol use. RESULTS: Only 5% of patients treated with FP100/salmeterol withdrew because of worsening asthma in the first 12 weeks; this compared with 7% in the FP250 group. All patients from a subset of sites continued in the study for an additional 12 weeks; only an additional 1% of patients treated with either FP100/salmeterol or FP250 withdrew because of worsening asthma. Secondary efficacy measures confirmed primary efficacy results. CONCLUSION: In patients requiring FP250 bid for asthma stability, FP100/salmeterol bid was steroid sparing, allowing a 60% reduction in the FP dose while maintaining overall asthma control. PMID- 12532098 TI - Rhinovirus-induced wheezing in infancy--the first sign of childhood asthma? AB - BACKGROUND: Although known as common causes of upper respiratory infections, rhinoviruses, enteroviruses, and corona-viruses are poorly studied as inducers of wheezing in infants, and their possible role in the development of childhood asthma has not been investigated. OBJECTIVE: The purposes of this study were to assess the occurrence of RV, enterovirus, and coronavirus infections in wheezing infants and to evaluate the association of these viral findings with early school age asthma. METHODS: In 1999, outcome in relation to asthma was studied in 82 of 100 initially recruited children who had been hospitalized for wheezing in infancy during the period 1992-1993. In 2000, etiologic viral studies regarding the index episode of wheezing were supplemented by rhinovirus, enterovirus, and coronavirus detection by RT-PCR from frozen nasopharyngeal aspirates in 81 of the children for whom adequate samples were available. Of these children, 66 had participated in the follow-up in 1999. RESULTS: Rhinoviruses were identified in 27 (33%) of the 81 children, enteroviruses in 10 (12%), and coronaviruses in none. Rhinoviruses were present as single viral findings in 22 (81%) of the 27 rhinovirus-positive cases, and rhinovirus infections were associated with the presence of atopic dermatitis in infancy. Enteroviruses were commonly encountered in mixed infections and had no association with atopy. As single viral findings, rhinoviruses were associated with the development of asthma (P =.047; odds ratio, 4.14; 95% CI, 1.02-16.77 versus rhinovirus-negative cases [by logistic regression adjusted for age, sex, and atopic dermatitis on entry)]. CONCLUSION: Our results present rhinoviruses as important inducers of wheezing even in infancy. The association with atopy and subsequent asthma calls for reevaluation of the role of rhinoviruses in the development of asthma. PMID- 12532099 TI - Increased oxidative stress and altered levels of antioxidants in asthma. AB - BACKGROUND: Reactive oxygen species might play an important role in the modulation of airway inflammation. There is evidence of an oxidant-antioxidant imbalance in asthma. Although several oxidants and antioxidants are likely to be involved, alterations in only limited parameters have been studied in isolation. OBJECTIVE: We investigated changes in a wide range of oxidants and antioxidants to create a comprehensive picture of oxidant-antioxidant imbalance. METHODS: In the peripheral blood of 38 patients with bronchial asthma and 23 control subjects, oxidative stress was measured in terms of superoxide anion generation by leukocytes, lipid peroxidation products, total nitrates and nitrites, total protein carbonyls, and total protein sulfhydrils in plasma. Antioxidant status was evaluated by measuring red blood cell superoxide dismutase and catalase activity, total blood glutathione, and glutathione peroxidase activity in red blood cells and leukocytes and total antioxidant capacity in plasma. RESULTS: Asthmatic patients showed increased superoxide generation from leukocytes, increased total nitrites and nitrates, increased protein carbonyls, and increased lipid peroxidation products and decreased protein sulfhydrils in plasma, indicating increased oxidative stress. They also showed increased superoxide dismutase activity in red blood cells and increased total blood glutathione and decreased glutathione peroxidase activity in red blood cells and leukocytes. Red blood cell catalase activity and the total antioxidant capacity of plasma were not altered. CONCLUSION: There are alterations in a wide array of oxidants and antioxidants, with balance shifting toward increased oxidative stress in asthma. Therapeutic augmentation of the antioxidant defenses might be beneficial. PMID- 12532100 TI - A comparison of the airway response to segmental antigen bronchoprovocation in atopic asthma and allergic rhinitis. AB - BACKGROUND: Patients with allergic asthma and those with allergic rhinitis (without asthma) share many immunopathologic features but differ in the presence of lower airway symptoms in response to antigen. OBJECTIVES: We sought to compare the airway inflammatory response to antigen in patients with atopic asthma and allergic rhinitis. METHODS: Segmental bronchoprovocation with saline or ragweed antigen was performed in 9 patients with atopic asthma and 9 patients with allergic rhinitis without asthma. The antigen dose used in segmental bronchoprovocation was 10% of the dose that caused a 20% decrease in FEV1 in response to inhalation challenge. Bronchoalveolar lavage (BAL) was performed from the saline- and antigen-challenged segments at 5 minutes and 48 hours after challenge. BAL fluid was analyzed for cell count and differential, distribution of lymphocytes, and concentration of soluble factors (histamine, IL-5, matrix metalloproteinase 9, tissue inhibitor of metalloproteinase 1, and fibronectin). In addition, BAL cells were cultured ex vivo, and IL-5, IFN-gamma, and IL-10 generation was measured. RESULTS: Antigen challenge led to similar patterns of cellular recruitment, mediator levels, and BAL cell cytokine generation in both groups; however, the dose of antigen required to promote comparable responses in the airway was significantly less in patients with asthma. CONCLUSION: These data suggest that the pattern of acute airway inflammation in response to allergen does not by itself explain antigen-induced lower airway obstruction and asthma symptoms. We speculate that other factors, such as increased airway sensitivity to allergen or preexisting airway injury and remodeling, might explain why patients with asthma and rhinitis differ in their clinical and physiologic response to antigen exposure. PMID- 12532101 TI - Omalizumab, a recombinant humanized anti-IgE antibody, reduces asthma-related emergency room visits and hospitalizations in patients with allergic asthma. AB - BACKGROUND: Prevention of serious asthma exacerbations is an important therapeutic goal in patients with asthma. OBJECTIVE: The purpose of this study was to investigate the effect of omalizumab (Xolair), a recombinant humanized monoclonal anti-IgE antibody, on the rate of serious exacerbations during long term therapy. METHODS: A pooled analysis was completed of 3 multicenter, randomized, double-blind, placebo-controlled phase III studies with omalizumab in adults/adolescents aged > or =12 years (n = 1071) and in children aged 6 to 12 years (n = 334) who required treatment with inhaled corticosteroids for allergic asthma. Rates of serious asthma exacerbations were computed and compared between omalizumab- and placebo-treated patients. Serious exacerbations were those leading to unscheduled outpatient visits, emergency room treatment, or hospitalization during 1 year of treatment. RESULTS: In all, 767 patients were treated with omalizumab (at least 0.016 mg/kg/IgE [IU/mL], administered subcutaneously every 4 weeks). Another 638 patients were treated with placebo. The rate of unscheduled, asthma-related outpatient visits was lower for the omalizumab-treated patients than for the placebo-treated patients (rate ratio [95% CI], 0.60 [0.44, 0.81]; P <.01), as were asthma-related emergency room visits (rate ratio [95% CI], 0.47 [0.24, 1.01]; P =.05). Importantly, hospitalizations for asthma were markedly reduced in patients receiving omalizumab (rate ratio [95% CI], 0.08 [0.00, 0.25]; P <.01). CONCLUSION: Omalizumab reduces the rate of serious asthma exacerbations and the need for unscheduled outpatient visits, emergency room treatment, and hospitalization in patients with moderate-to-severe allergic asthma. PMID- 12532102 TI - Is there an association between inhaled corticosteroids and bone density in postmenopausal women? AB - BACKGROUND: During the last decades, there has been increased concern about the association between oral corticosteroid (OC) therapy and osteoporosis. OBJECTIVE: The question currently discussed is whether inhaled corticosteroids (ICs) in recommended doses have any clinically relevant effects on bone mineral density (BMD). METHODS: We compared BMD in postmenopausal women exposed to corticosteroids only in inhaled form (IC group, n = 106) with that in women not exposed to corticosteroids (unexposed group, n = 674). BMD was also studied in 49 women exposed to OCs, intra-articular injections, or both in addition to ICs (OC group). The women were recruited from a population-based prospective cohort study. METHODS: We used a dietary survey, bone density measurement of the forearm, and a health questionnaire including an interview about past and present medication use. RESULTS: Mean BMD did not significantly differ between the IC group (0.434 g/cm2) and the unexposed group (0.429 g/cm2). The mean duration and dose of ICs was 8.2 +/- 5.03 years and 853 microg daily. Within the IC group, BMD stratified for cumulative dose of IC, duration, or current dose greater than or less than 1000 microg did not differ. BMD in the OC group was lower than that in the IC group (0.408 vs 0.434 g/cm2). CONCLUSION: No difference in BMD was noted between the IC group and unexposed control subjects, nor was any dose-response relationship observed between IC therapy and BMD. PMID- 12532103 TI - Receptor internalization is required for eotaxin-induced responses in human eosinophils. AB - BACKGROUND: CC chemokine receptor 3 (CCR3) is a major chemokine receptor involved in regulating eosinophil trafficking, and therefore the elucidation of ligand induced CCR3 events has important implications in understanding the biologic and pathologic properties of eosinophils. After ligand binding to CCR3, cellular signals include stimulatory (ie, calcium mobilization, actin polymerization, shape change, and chemotaxis) and inhibitory (ie, desensitization of the receptor) events. We have previously demonstrated that CCR3 undergoes rapid and prolonged ligand-induced internalization. OBJECTIVE: Here we explore the role of internalization in downstream cellular processes, including shape change, actin polymerization, calcium mobilization, and desensitization. METHODS: Peripheral blood-derived human eosinophils were pretreated with 2 mechanistically distinct inhibitors of internalization, sucrose and phenylarsine oxide, and functional responses were monitored. RESULTS: We first demonstrate that ligand-induced internalization is required for chemokine-induced eosinophil shape change. To define which signaling components upstream of eosinophil shape change required internalization, we next studied the role of internalization in calcium mobilization and actin polymerization. Sucrose and phenylarsine oxide pretreatment inhibited actin polymerization, implicating receptor internalization in this early response. In contrast, calcium mobilization was not inhibited by blockade of internalization. Finally, we were interested in testing the role of internalization in receptor desensitization. We first demonstrated that preincubation with eotaxin induced a dose-dependent desensitization in eotaxin induced eosinophil transepithelial migration. However, this phenomenon was not inhibited by blockade of internalization. CONCLUSION: These results establish that CCR3 internalization is critically involved in select eosinophil functional responses (ie, cellular shape change and actin polymerization) but not desensitization and calcium mobilization. PMID- 12532104 TI - Bradykinin increases the in vivo expression of the CXC chemokine receptors CXCR1 and CXCR2 in patients with allergic rhinitis. AB - BACKGROUND: Increased levels of bradykinin and IL-8 have been detected within the airways of individuals with active symptoms of allergic rhinitis and asthma. OBJECTIVE: We sought to investigate the in vivo effect of bradykinin on the expression of the IL-8 receptors CXCR1 and CXCR2 in nasal cells. METHODS: Nasal samples were obtained from patients with active allergic rhinitis; patients with mild, quiescent allergic rhinitis; and healthy control subjects. CXCR1 and CXCR2 mRNA expression in the nasal cells was measured by means of quantitative real time RT-PCR in baseline samples from all subjects, as well as in samples obtained after in vivo bradykinin challenge in healthy control subjects and patients with mild allergic rhinitis. CXCR1 and CXCR2 cell-surface expression was also assessed by means of flow cytometry in nasal epithelial cells at baseline and after ex vivo bradykinin challenge. RESULTS: No difference was seen in CXCR1 or CXCR2 mRNA expression between healthy control subjects and patients with quiescent allergic rhinitis at baseline; however, patients with active allergic rhinitis had increased baseline expression of both CXCR1 and CXCR2 mRNA. In vivo nasal bradykinin challenge significantly increased CXCR1 and CXCR2 mRNA expression in patients with quiescent allergic rhinitis but had no effect in healthy control subjects. Low levels of CXCR1 but not CXCR2 cell-surface expression was detected in nasal epithelial cells at baseline, and ex vivo bradykinin challenge induced CXCR2 cell-surface expression in nasal epithelial cells from patients with mild allergic rhinitis. CONCLUSION: This study demonstrates the in vivo regulation of chemokine receptors by means of bradykinin in human airway tissue in patients with allergic rhinitis. PMID- 12532105 TI - Myosin light chain kinase mediates eosinophil chemotaxis in a mitogen-activated protein kinase-dependent manner. AB - BACKGROUND: Eosinophil migration in the tissue is one characteristic feature of allergic diseases. The CC chemokine eotaxin plays a pivotal role in local accumulation of eosinophils. Myosin light chain kinase (MLCK) is known to regulate cytoskeletal rearrangement and cell motility by means of phosphorylation of myosin light chain (MLC). OBJECTIVE: We have previously shown that mitogen activated protein (MAP) kinases are important for eosinophil migration. In the present study we hypothesized that MLCK is downstream of MAP kinases, thereby linking the MAP kinase pathway to the activation of cytoskeletal components required for eosinophil chemotaxis. METHODS: Blood eosinophils were purified by using Percoll and anti-CD16 antibody-coated magnetic beads. We investigated the phosphorylation of MLCK and MLC by using the phosphorous 32-orthophosphates labeled eosinophils. The kinase activity of MLCK was determined by measuring the phosphotransferase activity for the MLCK-specific peptide substrate. The chemotaxis assay was performed in a 48-well Boyden microchamber. RESULTS: The phosphotransferase activity of MLCK for a substrate peptide was enhanced in eotaxin-stimulated eosinophils. We also found that eotaxin induced phosphorylation of MLCK in vivo in phosphorous 32-orthophosphate-labeled eosinophils. PD98059 (MAP/extracellular signal-regulated kinase inhibitor) or SB202190 (p38 MAP kinase inhibitor) abrogated the eotaxin-induced phosphorylation of MLCK. The phosphorylation of MLC was upregulated by eotaxin. Eosinophil chemotaxis was inhibited by means of pretreatment of the MLCK inhibitor ML-7. CONCLUSION: These results suggest that eotaxin regulates MLCK through both extracellular signal-regulated kinase 1/2 and p38 MAP kinase. MLCK activation is a critical step in the cytoskeletal rearrangements leading to eosinophil migration. PMID- 12532106 TI - Polymorphisms in the IL 18 gene are associated with specific sensitization to common allergens and allergic rhinitis. AB - BACKGROUND: Atopy has been linked to chromosome 11q22, a region that harbors the IL18 gene. IL-18 enhances IL-4/IL-13 production and induces IgE production that is directly associated with the pathogenesis of atopic disorders. OBJECTIVE: We sought to investigate whether genetic abnormalities in the regulatory regions of the IL18 gene predispose, in part, to susceptibility to atopy. METHODS: Among a white population of 105 families, the oldest child was examined with regard to atopic phenotypes and single-nucleotide polymorphisms (SNPs) within the IL18 gene. RESULTS: We have identified 5 novel SNPs in the IL18 gene (-920[t/c], 133[c/g], and -132[a/g] in promoter 2 [upstream of exon 2]; +179[c/a; Ser35Ser] in exon 4; and +486[c/t; Phe137Phe] in exon 6). Three SNPs are located in promoter 2, and one (-133[c/g]; nuclear factor 1 site) was significantly associated with high serum IgE levels (P =.001; odds ratio, 3.96) and specific sensitization to common allergens (P =.005; OR, 4.12). In addition, previously identified SNPs in exon 1 (+113[t/g] and +127[c/t]) and in promoter 1 (-137[g/c], GATA3 site) of the IL18 gene were significantly associated with high IgE levels (P < or =.005; OR, 3.27-3.90) and specific sensitization (P =.02 to.008; OR, 3.27 3.83). The SNP +127(g/t) in exon 1 was also a susceptibility locus for seasonal allergic rhinitis (P =.008; OR, 3.22). CONCLUSION: IL18 might be responsible for the linkage effects seen in the chromosomal region 11q22, which has been found previously with the phenotype "sensitization to mite allergen." Thus a suspected direct role of IL18 in the pathogenesis of atopy has been strengthened by the presence of 8 common SNPs in the promoter regions of IL18. PMID- 12532107 TI - The relevance of maternal immune responses to inhalant allergens to maternal symptoms, passive transfer to the infant, and development of antibodies in the first 2 years of life. AB - BACKGROUND: Asthma and other atopic diseases are strongly hereditary. Although the mother might play a special role, the mechanisms for such an effect are not clear. OBJECTIVE: We sought to investigate the influence of maternal immune responses to cat and mite allergens on (1) maternal symptoms, (2) the development of immune responses in the infant, and (3) the development of allergic disease during the first 3 years of life. METHODS: In sera from 465 mothers and 424 infants (cord blood), as well as in sera from 230 of the children at age 2 to 3 years, total IgE and IgE antibodies were measured by using CAP testing; IgG and IgG4 antibodies for the cat allergen Fel d 1 were measured by means of radioimmunoprecipitation. RESULTS: In both mothers and children, approximately 15% of sera contained IgG antibodies to Fel d 1 without IgE antibodies to cat. The strongest predictor of the maternal IgG antibody response was exposure to greater than 8 microg of Fel d 1/g of dust. Thus approximately 70% of children living in a house with a cat had received IgG antibodies from their mothers. In many cases the infant received IgG and IgG4 antibodies to Fel d 1 from a nonallergic mother. Maternal IgE antibodies were consistently associated with asthma; by contrast, the IgG antibody was not independently related to asthma but was related to rhinitis in the mothers (odds ratio, 2.6; 95% CI, 1.1-6.2) and to eczema in children. At age 3 years, 13 of 230 sera contained IgE antibodies to mite, but only 5 had IgE antibodies to cat. CONCLUSIONS: A significant proportion (approximately 15%) of mothers and children exposed to high concentrations of cat (but not mite) allergens have serum IgG antibodies without IgE antibodies. This IgG antibody is freely transferred to the infant and might influence IgG antibody production in the child. The results indicate the importance of understanding the mechanisms of tolerance to cats and raise questions about the independent role of the mother in the inheritance of allergy. PMID- 12532108 TI - There is no evidence of an inverse relationship between TH2-mediated atopy and TH1-mediated autoimmune disorders: Lack of support for the hygiene hypothesis. AB - BACKGROUND: The findings of an inverse relationship between T(H)1- and T(H)2 mediated disorders would provide strong empiric support to the hygiene hypothesis. OBJECTIVE: We sought to investigate the relationship between T(H)2 mediated atopic allergy and T(H)1-mediated autoimmune conditions in a nationally representative population. METHODS: We used logistic regression to analyze adult data from the Third National Health and Nutrition Examination Survey. Data on allergic and autoimmune disease history were available for 20,050 subjects, and data on atopy were available for a subsample of 7304 subjects. Atopy was defined by one or more positive skin prick test responses (wheal of > or =3 mm) to 10 common aeroallergens. Allergic disease was defined by patient reports of physician-diagnosed asthma, hay fever, or both. T(H)1-mediated autoimmune disease was defined by patient reports of physician-diagnosed type 1 diabetes mellitus, thyroid disorders, and/or rheumatoid arthritis. RESULTS: Adjusted for age and sex and taking into account the complex survey design, there was no relationship between atopy and a history of autoimmune disorders (adjusted odds ratio, 1.01; 95% CI, 0.61-1.67; P =.97). In contrast, physician diagnosis of allergic disorders was associated with a significant increased risk of physician-diagnosed autoimmune disorders (adjusted odds ratio, 1.67; 95% CI, 1.35-2.07; P <.001). CONCLUSION: We found no evidence of an inverse relationship between atopy and patient reports of physician-diagnosed common autoimmune disorders in the adult American population. Contrary to our initial hypothesis, reports of physician diagnosed common allergic disorders are positively associated with reports of physician-diagnosed autoimmune disorders, with this possibly being caused by ascertainment bias. These findings suggest that the T(H)1/T(H)2 paradigm might be an oversimplification. PMID- 12532110 TI - Chronic inflammation of the skin can be induced in IgE transgenic mice by means of a single challenge of multivalent antigen. AB - BACKGROUND: It is now widely accepted that IgE mediates immediate-type allergic response. However, the pathologic role of IgE is controversial in the chronic allergic inflammation observed in atopic diseases, such as asthma and atopic dermatitis. OBJECTIVE: We investigated the role of IgE in cutaneous allergic reactions by using 2 newly developed lines of antigen-specific IgE transgenic mice. METHODS: IgE transgenic mice were administered subcutaneously with corresponding antigens, and the subsequent ear swelling was measured. RESULTS: A single subcutaneous administration of TNP-conjugated ovalbumin (OVA) into the ears of nonimmunized mice carrying the TNP-specific IgE transgene elicited immediate-phase and late-phase ear swelling as expected, which peaked at 20 minutes and 8 hours later, respectively. Interestingly, however, 2 to 3 days after the antigen challenge, more intense ear swelling appeared. Its magnitude and duration were dependent on the valency of TNP in OVA, as well as the dose of TNP-OVA, and it lasted over 1 month when 100 microg of OVA conjugated with 11 molecules of TNP was given. Interestingly, administration of OVA to OVA-specific IgE transgenic mice elicited immediate-phase and late-phase ear swelling but not third-phase ear swelling. Massive infiltration of inflammatory cells was observed in the third-phase ear swelling of TNP-specific IgE transgenic mice. Cyclosporine A almost completely inhibited the third-phase ear swelling and cellular infiltration, whereas an antihistamine, cyproheptadine, did not show any significant effect on the third-phase reaction. CONCLUSION: These results indicate that IgE can trigger not only immediate-type hypersensitivity but also chronic allergic inflammation. Our findings highlight a novel immunopathologic role of IgE in chronic atopic disorders. PMID- 12532109 TI - TH2 dominance and defective development of a CD8+ dendritic cell subset in Id2 deficient mice. AB - BACKGROUND: Although the TH1/TH2 balance is important in many clinical situations, the regulatory mechanisms in vivo have not been well elucidated. OBJECTIVE: We sought to characterize the immunologic status of mice lacking Id2, an inhibitor of basic helix-loop-helix transcription factors. METHODS: We analyzed serum immunoglobulin levels, gene-expression profiles in the spleen, TH1/TH2 balance, and dendritic cell (DC) populations of Id2-/- mice. RESULTS: Serum levels of TH2-mediated IgG1 and IgE were increased more than 10-fold in Id2 /- mice without antigenic stimulation. Gene-expression analysis in Id2-/- splenocytes revealed enhanced expression of TH2-related genes, such as IL-4, and reduced expression of TH1-related genes, including IFN-gamma and IL-12. Intracellular cytokine staining also confirmed that Id2-/- splenic CD4+ T cells are substantially skewed to TH2 cells. However, Id2-/- naive CD4+ T cells differentiated into TH1 cells comparably with wild-type T cells under the appropriate culture conditions. Id2-/- mice displayed a selective and remarkable reduction of the CD8+ DC subset, which is known to induce preferential TH1 differentiation. CONCLUSION: Id2 is an indispensable regulator of the TH1/TH2 balance, possibly through the proper development of CD8alpha+ DCs, and could be a novel target to treat allergic diseases. PMID- 12532111 TI - Asymptomatic skin sensitization to birch predicts later development of birch pollen allergy in adults: a 3-year follow-up study. AB - BACKGROUND: The skin prick test is the allergologic test of choice, but asymptomatic skin sensitization to aeroallergens is common. However, no data in the literature describe the clinical phenotype of asymptomatic sensitized adults. OBJECTIVE: The purposes of this investigation were to provide a clinical characterization of skin test-positive subjects without symptoms and to ascertain the predictive values of common allergologic tests. METHODS: Asymptomatic adults with positive skin prick test results for birch (n = 15), nonatopic control subjects (n = 25), and birch pollen-allergic patients (n = 6) were followed through use of daily diary cards during 3 consecutive birch pollen seasons. At inclusion and at the 3-year follow-up visit, conjunctival and nasal challenges, intradermal late-phase reaction evaluation, and measurement of specific IgE were performed. RESULTS: Asymptomatic sensitized subjects defined a clinically significant phenotype between nonatopic and allergic subjects in terms of specific IgE levels and susceptibility to conjunctival provocation testing. Sixty percent (n = 9) of the asymptomatic sensitized subjects developed clinical allergy. This was associated with an initial birch skin prick test weal diameter of > or =4 mm, a positive conjunctival provocation test result, and specific IgE of > or =CAP class 2, as well as with the presence of other allergies. Specific IgE of > or =CAP class 2 was 87.5% predictive for allergy development, whereas a negative conjunctival provocation test result was 100% negatively predictive. Nasal provocation testing possessed no additional prognostic information. No changes in clinical phenotype were seen in nonatopic or birch-allergic subjects. CONCLUSION: Asymptomatic skin sensitization is a risk factor for later allergy development. At risk is any subject with target organ sensitivity, an elevated specific IgE level, and/or a skin prick test weal diameter of >4 mm. PMID- 12532112 TI - A double-blind, placebo-controlled immunotherapy dose-response study with standardized cat extract. AB - BACKGROUND: Allergen immunotherapy with doses of cat extract containing approximately 15 microg of the major allergen, Fel d 1, have been proved clinically effective in several double-blind, placebo-controlled studies. However, the maintenance doses used in allergy practice in the United States are often considerably less than this proven dose. OBJECTIVE: The purpose of this investigation was to determine whether maintenance immunotherapy with cat dander extract containing 0.6 microg or 3.0 microg of Fel d 1 was more effective than placebo and similar in efficacy to treatment with extracts containing 15.0 microg Fel d 1, immunologic parameters being used as the outcome. METHODS: Twenty-eight cat-allergic patients were randomly entered, 7 in each group, into a double blind, placebo-controlled comparison of the immunologic response to treatment with placebo or cat dander extract containing 0.6 microg, 3.0 microg, or 15.0 microg of Fel d 1. Maintenance doses were achieved in 8 visits over a period of 4 weeks through use of a cluster regimen; each subject then received 1 weekly maintenance injection before posttreatment measurements were made. The response to immunotherapy was assessed before immunotherapy and after the first weekly maintenance injection. Studies included responses to titrated skin prick tests to cat extract and an unrelated allergen and serum allergen-specific IgE and IgG4. Titrated nasal challenges were performed with cat extract; measurement of mRNA and secreted cytokines (IL-4, IL-5, and IFN-gamma) was done at 6 hours. Serum cytokines (IL-4, IL-5, IFN-gamma) were measured, and flow cytometric analysis of intracellular cytokines (IL-4, IL-5, IFN-gamma) was performed. Cat allergen stimulated lymphocyte proliferation was performed with measurement of cytokines in the supernatant (IL-4, IL-5, IFN-gamma). RESULTS: All 28 subjects completed the study. Significant and dose-dependent differences were encountered in the titrated skin prick tests (P =.008), the cat-specific IgG(4) (P =.01), and the reduction in CD4+/IL-4+ PBMCs on flow cytometry (P =.03). There were no significant differences between placebo and cat dander extract containing Fel d 1 0.6 microg. Both extracts containing 3.0 microg and 15.0 microg produced significant decreases in skin prick test sensitivity (P =.02 and P =.002, respectively). The extracts containing 3.0 microg and 15.0 microg produced significant increases in cat-specific IgG4 (P =.01 and P =.006, respectively). Only the 15.0-microg-per-dose extract produced a significant reduction in the percent of CD4+/IL-4+ PBMCs (P =.003). CONCLUSION: In this double-blind, placebo controlled study, a maintenance dose of cat dander extract containing 15.0 microg Fel d 1 produced the most consistent immunologic response. A dose of 3.0 microg reduced skin prick test sensitivity and increased cat-specific IgG4 but did not reduce the circulating CD4+/IL-4+ PBMCs, a change that is likely related to the clinically significant response to allergen immunotherapy. These findings suggest that a maintenance dose of 15.0 microg of Fel d 1 is most apt to be clinically effective for allergen immunotherapy. PMID- 12532113 TI - Eighteen-month outcomes of house dust mite avoidance and dietary fatty acid modification in the Childhood Asthma Prevention Study (CAPS). AB - BACKGROUND: Observational studies have linked house dust mite (HDM) exposure and dietary fatty acid intake with asthma in childhood. However, definitive evidence of their role in the etiology of asthma requires a randomized controlled trial. OBJECTIVE: We hypothesized that the incidence of asthma and allergy in high-risk children would be reduced by avoidance of HDM allergens, supplementation with omega-3 fatty acids, or the combination of these strategies. We present the results of an interim analysis reporting outcomes assessed at 18 months. METHODS: A total of 616 pregnant women were randomized to an HDM avoidance intervention, comprising the use of impermeable mattress covers and an acaricide or control and the use of an oil supplement, margarines, and cooking oils containing high levels of omega-3 fatty acids or control. Atopic status was measured by skin prick testing. Symptoms, diagnoses, and medication histories were elicited by means of parental interviews. RESULTS: The diet intervention resulted in a 9.8% absolute reduction (95% CI, 1.5-18.1; P =.02) in the prevalence of any wheeze and a 7.8% absolute reduction (95% CI, 0.5-15.1, P =.04) in prevalence of wheeze of >1 week, but it had no effect on serum IgE, atopy, or doctors' diagnosis of asthma. The HDM avoidance intervention did not affect these outcomes but was associated with a lower use of oral steroids. CONCLUSION: Increasing dietary omega-3 fatty acids might have a beneficial effect on the prevalence of wheeze during the first 18 months of life. Follow-up to age 5 years, when the effect of the interventions on asthma risk will be assessed, is underway. PMID- 12532114 TI - Effect of mattress and pillow encasings on children with asthma and house dust mite allergy. AB - BACKGROUND: House dust mite (HDM) allergy is a frequent cause of allergic asthma in children. Reduction of exposure seems to be the most logical way to treat these patients. OBJECTIVE: Our aim was to investigate whether mattress and pillow encasings resulted in an effective long-term control of HDM allergen levels, thereby reducing the need for asthma medication in children with asthma and HDM allergy. METHODS: In a prospective, double-blind, placebo-controlled study 60 children (age range, 6-15 years) with asthma and HDM allergy were randomized to active (allergy control) or placebo mattress and pillow encasings. After a 2-week baseline period, follow-up was performed every 3 months for 1 year. During the entire study period, the dose of inhaled steroids was tapered off to the lowest effective dose according to well-defined criteria. RESULTS: Fifty-two patients completed the trial, and 5 were excluded, leaving data from 47 children (26 in the active treatment group and 21 in the placebo group) for analysis. A significant perennial reduction in HDM allergen concentrations was seen only for the active treatment group. Also, a significant decrease in the dose of inhaled steroids (mean, 408 to 227 microg/d; P <.001) was found for the active treatment group only, with significant differences between groups after 9 and 12 months. After 1 year, the dose of inhaled steroids was reduced by at least 50% in significantly more children in the active treatment group than in the placebo group (73% vs 24%, P <.01). CONCLUSION: Encasing of mattresses and pillows resulted in a significant long-term reduction in HDM allergen concentrations in mattresses and in the need for inhaled steroids in children with asthma and HDM allergy. PMID- 12532115 TI - Cotton fluffs as latex allergen carriers in a glove factory. AB - BACKGROUND: Latex allergens released from gloves and bound to airborne cornstarch powder have been shown to cause respiratory allergy in health care workers. OBJECTIVE: Cornstarch from powdered latex gloves is a well-known carrier of airborne allergens. It should be investigated whether cotton fluffs used to lubricate household and industrial gloves also function as airborne carriers for latex allergens. METHODS: Latex gloves, cotton fluffs with or without latex glove contact, dipping water from the glove cooling tank, a dust sample from the floor, and 10 samples from area dust samplers were analyzed for their allergen content by using a CAP inhibition method. Furthermore, 84% (119/142) of the working group participated in a cross-sectional study involving a questionnaire and measurement of latex-specific IgE by means of CAP inhibition. RESULTS: Latex gloves, area dust samples from latex glove production sites, and the floor dust sample contained high concentrations of latex allergens. Cotton fluffs were free of allergens before being in contact with gloves but showed considerable allergen concentrations after their removal from gloves. The dipping water of the cooling tank also contained latex allergens. Of the examined employees, 12.6% had IgE mediated sensitization, and a third of the sensitized subjects (ie, 4% of the total group) reported work-related hypersensitivity reactions. CONCLUSIONS: Cotton fluffs bind latex allergens and behave like cornstarch as airborne allergen carriers causing conjunctivitis and allergic respiratory disorders. PMID- 12532116 TI - Long-term treatment with aspirin desensitization in asthmatic patients with aspirin-exacerbated respiratory disease. AB - BACKGROUND: Aspirin desensitization treatment is an option to decrease disease activity and reduce the need for systemic corticosteroids in patients with aspirin-exacerbated respiratory disease (AERD). OBJECTIVE: This study was designed to determine whether the clinical courses of patients with AERD improved as early as 6 months after starting aspirin desensitization and to compare this with follow-up evaluations after at least a year. METHODS: Between 1995 and 2000, 172 patients with AERD were admitted to our General Clinical Research Center, were desensitized to and treated with aspirin, were discharged to their home communities, and participated in follow-up interviews and written assessments of their clinical courses. RESULTS: By the first 6 months of aspirin treatment, there were significant reductions in sinus infections and numbers of short courses of prednisone and improvements in sense of smell and general assessment of nasal-sinus and asthma symptoms (P <.0001). These results persisted for 1 to 5 years (P <.0001). Mean prednisone doses decreased from 10.8 mg/d to 8.1 and 3.6 mg/d at 6 months and greater than 1 year, respectively. Of the 172 patients, 24 (14%) discontinued aspirin treatment because of side effects, and 115 (67%) responded to aspirin treatment. After eliminating those who discontinued aspirin treatment because of side effects, the improvement rate was 115 (78%) of 148 patients. Of the 126 patients who completed a year or more of aspirin treatment, 110 (87%) experienced improvement. CONCLUSION: Aspirin desensitization followed by daily aspirin is efficacious by at least the first 6 months of treatment and continues to be effective for up to 5 years of follow-up. PMID- 12532117 TI - Prevalence, severity, and natural history of jack jumper ant venom allergy in Tasmania. AB - BACKGROUND: The jack jumper ant (Myrmecia pilosula) is responsible for greater than 90% of Australian ant venom allergy. However, deaths have only been recorded in the island of Tasmania. OBJECTIVES: We sought to determine the prevalence, clinical features, natural history, and predictors of severity of M pilosula sting allergy in Tasmania. METHODS: We performed a random telephone survey supported by serum venom-specific IgE analysis, review of emergency department presentations, and follow-up of allergic volunteers. RESULTS: M pilosula, honeybee (Apis mellifera), and yellow jacket wasp (Vespula germanica) sting allergy prevalences were 2.7%, 1.4%, and 0.6% compared with annual sting exposure rates of 12%, 7%, and 2%, respectively. Similarly, emergency department presentations with anaphylaxis to M pilosula were double those for honeybee. M pilosula allergy prevalence increased with age of 35 years or greater (odds ratio [OR], 2.4) and bee sting allergy (OR, 16.9). Patients 35 years of age or older had a greater risk of hypotensive reactions (OR, 2.9). Mueller reaction grades correlated well with adrenaline use. During follow-up, 79 (70%) of 113 jack jumper stings caused anaphylaxis. Prior worst reaction severity predicted the likelihood and severity of follow-up reactions; only 3 subjects had more severe reactions. Venom-specific IgE levels and other clinical features, including comorbidities, were not predictive of severity. CONCLUSIONS: Sting allergy prevalence is determined by age and exposure rate. M pilosula sting exposure in Tasmania is excessive compared with that found in mainland Australia, and there is a high systemic reaction risk in allergic people on re-sting. Prior worst reaction severity (Mueller grade) and age predict reaction severity and might be used to guide management. PMID- 12532118 TI - Exhaled nitric oxide and atopy in children. PMID- 12532119 TI - Anaphylaxis to rice by inhalation. PMID- 12532120 TI - Clinical and immunologic reactivity to aeroallergens in "intrinsic" atopic dermatitis patients. PMID- 12532121 TI - A TH2 cytokine inhibitor for airway inflammation in mild asthma. PMID- 12532122 TI - Fixed drug eruptions caused by tonic water. PMID- 12532123 TI - Risk factor for latex allergy in 54 children with atopy and latex sensitization. PMID- 12532124 TI - Community pharmacists as medication educators. PMID- 12532125 TI - Atopic dermatitis and the clinical effect of house dust mite avoidance. PMID- 12532126 TI - Cost-effectiveness of pediatric asthma treatments. PMID- 12532127 TI - Clinical significance of bronchoalveolar cell profiles in childhood asthma. PMID- 12532128 TI - Coca and Cooke on the classification of hypersensitiveness. PMID- 12532131 TI - Recent advances in the repair of groin herniation. PMID- 12532132 TI - Evaluation of chiropractic management of pediatric patients with low back pain: a prospective cohort study. AB - BACKGROUND: Recent epidemiologic studies have estimated that the lifetime prevalence of low back pain (LBP) in children is approximately 50%, with almost 15% of children experiencing frequent or continual pain. A literature search revealed no published studies addressing conservative treatment of childhood LBP. OBJECTIVE: To describe chiropractic management of LBP in patients between the ages of 4 and 18 years, as well as outcomes and factors associated with the outcomes. METHODS: Prospective cohort study of consecutive pediatric patients with LBP seeing randomly selected chiropractors within the cities of Calgary, Alberta, and Toronto, Ontario, Canada. Follow-up data collection included the type and extent of treatment rendered and its outcome, which was measured with a 5-point subjective rating scale and a self-report pediatric visual analogue scale. RESULTS: Fifteen chiropractors provided data on 54 consecutive pediatric patients with LBP. The average age of the patients was 13.1 years, 57% were male, 61% were acute, with 47% attributing onset to a traumatic event (most commonly sports-related); 24% reported an episode duration of greater than 3 months. Almost 90% of cases presented with uncomplicated mechanical LBP, most frequently diagnosed as lumbar facet dysfunction or subluxation. Patients were managed with manipulation, with a minority (7.7%) receiving some form of active management. "Important" improvement was seen in 62% and 87% on the visual analogue and subjective scales, respectively, within a 6-week course of management (Kaplan Meier survival analysis). Patients with chronic LBP were less likely to respond within the median number of treatments (relative risk = 2.1). CONCLUSIONS: Patients responded favorably to chiropractic management, and there were no reported complications. Future investigations should establish the natural history and compare chiropractic management with other forms of treatment to gain knowledge about the effectiveness of chiropractic in managing pediatric LBP. PMID- 12532133 TI - Chiropractor's use of radiography in Switzerland. AB - OBJECTIVE: In this article, we investigate the use of diagnostic radiology by chiropractors in Switzerland, with the aim of determining their contribution to the annual radiation collective dose. METHODS: We approached 138 chiropractors possessing radiologic equipment and asked them to provide, among other information, the frequency of radiographic examinations. The dose associated with each type of radiographic examination was established separately. The collective dose was determined by convolution of frequency and dosimetric information. RESULTS: The number of chiropractic radiographic examinations performed in Switzerland in 1998 was 60,000, mainly spinal and pelvic examinations. The associated annual collective dose was found to be 144 Sv (an annual effective dose of 20 microSv per capita). The chiropractic contribution to the total dose due to conventional radiography in Switzerland is about 6%. CONCLUSIONS: Considering the number of chiropractors in Switzerland (less than 200), their contribution to the collective dose is relatively high. This is due to the high effective dose associated with the main types of examinations performed (eg, lumbar spine). It is therefore necessary to develop and apply guidelines for the use of this type of radiographic examination, as well as quality control programs, in order to optimize the radiographic technique and hence reduce the doses. PMID- 12532134 TI - Empowerment of chiropractic faculty: a profile in context. AB - BACKGROUND: The primary resource base and core human capital in chiropractic education is found among its faculty. The chiropractic profession depends on a motivated faculty for continuous quality improvement and innovation in areas of curriculum, scholarship, and practice. Empowerment has been associated with increased intrinsic motivation. Assessments of attitudes of faculty in terms of empowerment, a set of cognitions created by the work environment, may inform executive decision making related to development of a productive professional work environment. OBJECTIVE: The goal of this exploratory study is to provide an initial understanding of empowerment among faculty in the organizational context of chiropractic education, to construct a profile of perceived faculty empowerment, and to inquire into potential associations between perceived empowerment and faculty demographic and workplace characteristics. STUDY DESIGN: This was a full faculty survey using descriptive statistics and multivariable analysis. METHODS: Surveys were distributed to full- and part-time faculty working in the United States and Canada. The survey was composed of Spreitzer's multidimensional measure of psychologic empowerment and additional items designed to measure faculty demographics and workplace variables including academic rank, years at the institution, years in higher education, sex, age, area of assignment, employment status, and academic rank. RESULTS: More than 54% of the study population (N = 609) completed and returned the instrument. The respondents were typically male (68.4%) and employed full-time (81.6%). Almost half (47.5%) of the respondents were assigned to the area of patient care at their institution. Area of assignment and employment status emerged as important variables for explaining the variance in dependent variable scores. CONCLUSION: The findings of this research provide a basic profile and some initial perspectives of empowerment in the context of the work environment of the chiropractic profession's learning institutions. PMID- 12532135 TI - Comparative analysis of low-back loading on chiropractors using various workstation table heights and performing various tasks. AB - BACKGROUND: There is epidemiologic evidence that chiropractors are a high-risk group for low-back disorders. However, to date there are no known biomechanical studies to determine whether their workstations may be a contributing factor. OBJECTIVE: To investigate whether chiropractors' workstation table height or the tasks they perform make them susceptible to low-back strain. As well as investigating low-back strain, a screening was performed to determine whether chiropractors' upper extremities were at risk for undue strain as workstation table height was varied. STUDY DESIGN: Experimental pilot study. SETTING: A university ergonomic laboratory. METHODS: An adjustable manipulation table was set at 3 different heights: 465 mm, 665 mm and 845 mm. Each of the 7 volunteer chiropractors were fitted with a triaxial electrogoniometer and were videotaped and photographed for analysis while performing spinal manipulation to the cervical, thoracic, and lumbar spine of a volunteer patient at each workstation table height. Two biomechanical models, one static and one dynamic, were used to record the dependent variables. A screening of various upper extremity variables was also performed with the static model. RESULTS: For the subjects under study, a significant difference was found for the variables maximum sagittal flexion, disk compression force, and ligament strain as table height was varied. For the lumbar and thoracic manipulation tasks, the medium table height (655 mm) was found to create the least low-back strain. For the cervical manipulation task, the high table height (845 mm) was found to be the least straining on the low back. The low height table (465 mm) was the most straining for all tasks. Upper extremities were not significantly affected by changes to table height. Significant differences were found for the task performed for axial rotational velocity, disk compression force, ligament strain, maximum sagittal flexion, dominant (right) elbow moment, and dominant (right) shoulder moment variables. There was no significant interaction between table height and task performed. CONCLUSION: Workstation table height was found to have a significant effect on low-back load of subjects under study. The results of this study demonstrate an overall unacceptably high amount of sagittal flexion, ligament strain, and disk compression force on the chiropractor subjects in the tasks performed. PMID- 12532136 TI - Objective manual assessment of lumbar posteroanterior stiffness is now possible. AB - OBJECTIVE: To evaluate the reliability and criterion-related validity of a revised version of the reference-based protocol for manually assessing lumbar posteroanterior (PA) stiffness. With the revised protocol, a therapist matched the stiffness of the spine to 1 of 11 stiffness references provided by a mechanical device. SUBJECTS: Two physiotherapists and 41 asymptomatic subjects participated. METHODS: Subjects' PA stiffness was assessed by the therapists and the Stiffness Assessment Machine(criterion measure) under standardized conditions. The therapists pressed on the subject's spine and then the mechanical device, the task being to match the stiffness of the back to 1 of the 11 stimuli provided by the device. Each of the 11 stiffness stimuli represented a point on the -5 to +5 stiffness scale. Interrater reliability was evaluated with 2-way analysis of variance (ANOVA) intraclass correlation coefficient for a single rating (ICC [2,1]) and the SEM, whereas criterion-related validity was evaluated with the Pearson product moment correlation (r), including the Fleiss correction. RESULTS: The ICC (2,1) was 0.78 (SEM 0.56), and the criterion-related validity reached 0.74 (Fleiss correction 0.86). CONCLUSIONS: This research shows that therapists can accurately judge asymptomatic subjects' spinal stiffness using a matching task. We believe that this method may allow therapists to quantify lumbar PA stiffness in clinical practice. PMID- 12532137 TI - Predicting academic success in the first year of chiropractic college. AB - PURPOSE: To determine if any existent preadmission academic or personal variables predict academic success in the first year of the Palmer College of Chiropractic West (PCCW) program. PARTICIPANTS: One hundred ninety-two students at PCCW who had completed the first year of the program. METHODS: One-way analysis of variance and stepwise linear multiple regression. RESULTS: Men had a significantly higher mean matriculating grade point average (MatGPA) than women, but no such relationship existed in the cumulative year-1 GPA (Y1GPA). There was no statistically significant difference in MatGPA for students possessing a degree compared to those without a degree, but degree-holding students had a significantly higher Y1GPA. There was no statistically significant difference in MatGPA for students born in English-speaking countries compared to students born in non-English-speaking countries, but those born in English-speaking countries had a significantly higher Y1GPA. MatGPA, physics GPA, and chemistry GPA provided the strongest regression model, eliciting an R(2) value of 0.327. CONCLUSIONS: Student characteristics on entering PCCW may help predict student performance in the first academic year. A relatively strong and statistically significant prediction model for Y1GPA (R(2) = 0.327) exists for PCCW. Used in conjunction with other available empirical data, this regression model may allow the institution to make more informed decisions when selecting students for admission. PMID- 12532138 TI - Is chiropractic evidence based? A pilot study. AB - OBJECTIVE: To calculate the proportion of care delivered in a chiropractic practice supported by good-quality clinical trials. DESIGN: Retrospective survey. METHODS: Data were collected from patient files relating to 180 consecutive patient visits in a suburban chiropractic practice in northern Spain. Each patient's presenting complaint was paired with the chiropractor's chosen primary intervention. Based on a literature review (Medline, Mantis, and nonautomated searches of local medical libraries), each presenting complaint-primary intervention pairing was categorized according to the level of supporting evidence as follows: Category I, intervention based on good quality clinical trial evidence; Category II, intervention based on poor-quality or no clinical trial evidence. To distinguish between good- and poor-quality clinical trials, studies were critically appraised and assigned quality scores. RESULTS: Of the 180 cases surveyed, 123 (68.3%) (95% CI, 61.5%-75.1%) were based on clinical trials of good methodologic quality (Category I). Only 57 (31.7%) (95% CI, 24.9% 38.5%) of the cases were based on poor-quality or no clinical trial evidence (Category II). CONCLUSIONS: When patients were used as the denominator, the majority of cases in a chiropractic practice were cared for with interventions based on evidence from good-quality, randomized clinical trials. When compared to the many other studies of similar design that have evaluated the extent to which different medical specialties are evidence based, chiropractic practice was found to have the highest proportion of care (68.3%) supported by good-quality experimental evidence. PMID- 12532139 TI - Is cervical spinal manipulation dangerous? AB - OBJECTIVE: Concern about cerebrovascular accidents after cervical manipulation is common. We report a case of cerebrovascular infarction without sequelae. CLINICAL FEATURES: A 39-year-old man with nonspecific neck pain was treated by his general practitioner with cervical manipulation. INTERVENTION AND OUTCOME: This immediately elicited severe headache and neurologic symptoms that disappeared completely within 3 months despite permanent signs of a complete left-sided cerebellar infarction on computed tomography and magnetic resonance imaging. At 7 year follow-up the patient was fully employed, and repeated magnetic resonance imaging still showed infarction of the left cerebellar hemisphere. However, the patient remained completely free of neurologic symptoms, and color duplex ultrasonography showed normal cervical vessels, including patent vertebral arteries. CONCLUSION: It appears that the risk of cerebrovascular accidents after cervical manipulation is low, considering the enormous number of treatments given each year, and very much lower than the risk of serious complications associated with generally accepted surgery. Provided there is a solid indication for cervical manipulation, we believe that the risk involved is acceptably low and that the fear of serious complications is greatly exaggerated. PMID- 12532140 TI - Missed lumbar transverse process fractures in a high school football player. PMID- 12532141 TI - Unsubstantiated claims in patient brochures from the largest state provincial, and national chiropractic associations and research agencies. PMID- 12532143 TI - Rating specific chiropractic technique procedures for common low back conditions. PMID- 12532145 TI - Rating specific chiropractic technique procedures for common low back conditions. PMID- 12532147 TI - Two forms of disordered eating in obesity: binge eating and night eating. AB - OBJECTIVE: Binge eating disorder (BED) and the night eating syndrome (NES) have been linked to obesity. This review summarizes their characteristics, implications of their diagnoses and treatment outcomes. METHOD: Selective review of the literature on BED and NES. RESULTS: BED was proposed as a distinctive disorder on the basis of two large multisite studies in the early 1990s. It is associated with more severe and earlier onset of obesity, earlier onset of dieting and greater psychopathology. It shows large placebo responses and reduction of bingeing in patients on waiting-list controls. Traditional weight reduction programs reduce bingeing at least as well as psychological treatments designed for this purpose. NES is a stress-related eating, sleeping and mood disorder that is associated with disordered neuroendocrine function. It follows a characteristic circadian pattern and has responded to an agent that enhances serotonin function. CONCLUSIONS: BED responds well to weight reduction programs. It is proposed that this diagnosis be used as a marker for psychological problems that deserve treatment in their own right. NES is an eating, sleep, and mood disorder with distinctive behavioral and neuroendocrine characteristics. Studies of treatment for NES are in their infancy but selective serotonin reuptake inhibitors (SSRI) show promise. PMID- 12532148 TI - Relationship between high plasma leptin concentrations and metabolic syndrome in obese pre-pubertal children. AB - OBJECTIVE: To evaluate the relationship between serum leptin levels and metabolic syndrome, fasting insulin level and anthropometric index in obese pre-pubertal children. DESIGN: A cross-sectional study was carried out on obese children. SUBJECTS: A study was made of 41 obese children (aged 6-9 y) and the same number of non-obese children (control group), matched by age and sex. METHODS: Body mass index (BMI), waist/hip ratio (WHR) and blood pressure were determined in each child. Serum leptin, glucose, insulin, lipid profile, sex hormone binding globulin (SHBG), plasminogen activator inhibitor-1 (PAI-1), tissue-plasminogen activator (t-PA) and fibrinogen were all measured. RESULTS: The serum leptin level was significantly higher in obese children (15.47 vs 4.73 ng/ml). In the obese group, leptin showed a positive correlation with BMI (P<0.001), insulin (P<0.001), triglycerides (P<0.05), PAI-1 (P<0.05) and t-PA (P<0.05), and correlated negatively with SHBG (P<0.01), apolipoprotein A-I (P<0.05) and high density lipoproteins cholesterol (HDL-C) (P<0.05). Corrected for BMI and WHR, leptin (P partial=0.002) is only an independent predictive factor for basal insulin. Using multivariant regression analysis, only insulin (P partial=0.003) and BMI (P partial=0.018) were independent predictive factors for leptin. CONCLUSION: For this age group, high leptin resistance may be another component of metabolic syndrome, and may be involved in its etiopathogenesis. The involvement of leptin in this syndrome may be indirect, modulating the insulin's action. PMID- 12532149 TI - Body composition and metabolic factors in obese children and adolescents. AB - OBJECTIVE: Body composition is associated with metablic factors in adults; however, data are limited regarding obese children. This study was undertaken to assess body composition, regional fat distribution, and metabolic factors in obese 6-18-y-old children and adolescents. DESIGN: Cross-sectional assessment. SUBJECTS: Thirty-six obese children and adolescents, (mean+/-s.e.m.) age 11.8+/ 0.5 y, BMI 34.1+/-1.2 kg/m(2). MEASUREMENTS: Body composition was assessed by dual energy X-ray absorptiometry and computerized tomography. Fasting insulin, glucose and leptin levels, and the homeostasis model assessment of insulin sensitivity (HOMA-IR) were assessed. RESULTS: The girls had significantly lower glucose levels than the boys. The ethnic group differences (African American children vs white children) in fat mass, total CT fat, subcutaneous CT fat, insulin level, leptin level, and higher HOMA-IR were not significant after adjusting for age or pubertal stage. These differences in abdominal fat and subcutaneous abdominal fat were also not independent of total body fat or BMI. No ethnic group differences in visceral abdominal fat were noted. Insulin level and HOMA IR were associated with leptin level (independent of fat mass) and fat mass. Leptin level was associated with fat mass, total CT fat, and subcutaneous CT fat; however the associations between the CT fat measures and leptin were not independent of total body fat mass. CONCLUSIONS: Neither visceral abdominal fat, subcutaneous abdominal fat, insulin levels, or insulin resistance differed by ethnic group when adjusted for age or pubertal status. This contrasts with findings in adults and non-obese children which suggest lower levels of visceral fat and higher insulin levels and insulin resistance in African American children and adolescents. PMID- 12532150 TI - Severe insulin resistance contrasting with mild anthropometric changes in the adipose tissue of HIV-infected children with lipohypertrophy. AB - BACKGROUND: The HIV-associated lipodystrophic syndrome (HIV-LDS) combines redistribution of fat mass with insulin resistance and hyperlipidemia. We have previously reported that HIV-LDS prevails in children in a comparable pattern as in adults. The metabolic activity itself of the lipodystrophic adipose tissue in HIV infection has been poorly studied. AIM AND METHODS: To assess in situ the insulin sensitivity of the lipohypertrophic subcutaneous abdominal adipose tissue using the microdialysis technique in HIV-infected children. Insulin sensitivity, assessed by the inhibition of glycerol release, was measured in the abdominal subcutaneous adipose tissue during a standard oral glucose tolerance test (OGTT) in six HIV-infected children under multi-therapy with abdominal lipohypertrophy (supra-iliac skinfold thickness >97th percentile) (HIV/LH+), in six obese children (obese group) and in eight HIV-infected children without lipodystrophy (HIV/LH-). RESULTS: Glucose tolerance was normal in all subjects. Mean insulin areas under the curve (IAUC) were significantly higher in the obese and HIV/LH+ groups than in HIV/LH- (8769+/-5429, 8161+/-4552 and 3618+/-2222 mU min l(-1), respectively; P=0.04 for the three groups comparison by the Kruskal-Wallis test), reflecting insulin resistance in the two former groups independent of a significant difference in percentage fat mass (37.2+/-4.7, 22.8+/-10.9 and 20.7+/ 7.1%, respectively; P= 0.006). The crude inhibition of glycerol release, expressed as the relative change in dialysate glycerol concentration between baseline and 120 min, was not statistically different between the three groups (14% in obese, -38+/-14% in HIV/LH+ and -51+/-17% in HIV/LH- groups; P=0.3). The inhibition of glycerol release with respect to the circulating insulin level (expressed by IAUC) was similar in HIV/LH+ and obese groups (-6+/-5 x 10(-3) and 7+/-5 x 10(-3) l mU(-1) min(-1), respectively, P=0.4 for two-groups comparison by the U-Mann-Whitney test) and four-fold less than in the HIV/LH- group (-24+/-25 x 10(-3) l mU(-1) min(-1); P=0.02). CONCLUSION: These data argue in favor of insulin resistance in the adipose tissue of lipohypertrophies associated with HIV infection. PMID- 12532151 TI - Cell-permeable ceramides increase basal glucose incorporation into triacylglycerols but decrease the stimulation by insulin in 3T3-L1 adipocytes. AB - OBJECTIVE: To investigate mechanisms for the regulation of glucose incorporation into triacylgycerols in adipocytes by ceramides, which mediate some actions of tumour necrosis factor-alpha (TNFalpha). DESIGN: The effects of C(2)- and C(6) ceramides (N-acetyl- and N-hexanoyl-sphingosines, respectively) on glucose uptake and incorporation into triacylglycerols and pathways of signal tansduction were measured in 3T3-L1 adipocytes. RESULTS: C(6)-ceramide increased basal 2 deooxyglucose uptake but decreased insulin-stimulated uptake without changing the EC(50) for insulin. Incubating 3T3-L1 adipocytes from 2 to 24 h with C(2) ceramide progressively increased glucose incorporation into the fatty acid and especially the glycerol moieties of triacylglycerol. These effects were accompanied by increased GLUT1 synthesis resulting from ceramide-induced activation phosphatidylinositol 3-kinase, ribosomal S6 kinase and mitogen activated protein kinase. C(2)-ceramide also increased p21-activated kinase and protein kinase B activities. However, C(2)-ceramide decreased the insulin stimulated component of these signalling pathways and also glucose incorporation into triacylglycerol after 2 h. CONCLUSIONS: Cell-permeable ceramides can mimic some effects of TNFalpha in producing insulin resistance. However, ceramides also mediate long-term effects that enable 3T3 L1 adipocytes to take up glucose and store triacylglycerols in the absence of insulin. These observations help to explain part of the nature and consequence of TNFalpha-induced insulin resistance and the control of fat accumulation in adipocytes in insulin resistance and obesity. PMID- 12532152 TI - Dominant negative alpha-subunit of farnesyl- and geranylgeranyl-transferase I inhibits insulin-induced differentiation of 3T3-L1 pre-adipocytes. AB - OBJECTIVE: To investigate whether the expression of a dominant negative (DN) farnesyl- and geranygeranyl-transferase I (FTase/GGTase I) alpha-subunit in 3T3 L1 pre-adipocytes can inhibit insulin's ability to induce differentiation. DESIGN: 3T3-L1 pre-adipocytes were stably transfected with vector alone or vector expressing a mutated DN FTase/GGTase I alpha-subunit (S60A)(S62A) and incubated in serum-free medium in the absence and presence of insulin. MEASUREMENTS: Various assays were performed to determine the effect of DN FTase/GGTase I alpha subunit expression in 3T3-L1 pre-adipocyte on insulin-induced DNA synthesis, cell count, phosphorylation of the FTase/GGTase I alpha-subunit, FTase and GGTase I activity, amounts of prenylated p21Ras and RhoA, phosphorylation of MAP kinase and Akt, and differentiation to mature fat cells. RESULTS: Expression of DN FTase/GGTase I alpha-subunit inhibited insulin's ability to increase DNA synthesis, cell count, FTase and GGTase I activity, amounts of prenylated p21Ras and RhoA, and magnitude of phosphorylation of MAP kinase. Expression of DN FTase/GGTase I alpha-subunit in 3T3-L1 pre-adipocytes was without effect on insulin-induced Akt phosphorylation. CONCLUSION: Expression of DN FTase/GGTase I alpha-subunit inhibits insulin-induced differentiation of 3T3-L1 pre-adipocytes to mature adipocytes, and thus could indicate potential therapeutic avenues to assuage the deleterious effects of obesity and type 2 diabetes. PMID- 12532153 TI - Leptin reduces food intake but does not alter weight regain following food deprivation in the rat. AB - OBJECTIVE: When animals are allowed free access to food following an extended period of food restriction, body weight is steadily restored to the pre-food restriction level, ie to a specific body weight 'set-point'. We tested the proposition that leptin is used as a signal by the brain to regulate body weight 'set-point'. To this end, we determined whether long-term leptin infusion in rats would prevent the normal weight regain after food restriction. METHODS: Male Sprague-Dawley rats received leptin (leptin-treated) or saline (vehicle-treated) by intravenous infusion. After a 2 week run-in period, food intake was adjusted to 50% of each individual's normal intake for 12 days. Two days prior to the return of unlimited access to food, one group of animals received continuous leptin infusion at 3 micro g/h for the next 14 days. Blood samples taken from the tail vein were used to determine leptin concentrations. A third group of animals that did not undergo food restriction but received saline infusion served as control. As leptin acts in the brain to modulate neuropeptide Y (NPY) levels, hypothalamic NPY content was measured at the end of the study. RESULTS: Food restriction to 50% normal intake for 12 days induced a 20% weight loss and significant reductions in plasma leptin compared with non-restricted control rats (0.5+/-0.1 vs 2.6+/-0.4 ng/ml, P<0.05). Intravenous infusion of leptin increased leptin concentrations four-fold compared with vehicle-treated animals (9.5+/-1.3 vs 2.2+/-0.4 ng/ml, P<0.05). The infusion of leptin attenuated the increase in daily food intake after free access to food was resumed (P<0.05 at 4, 6 and 8 days). Despite this, both groups of previously restricted rats had regained the same amount of weight after 12 days of ad libitum feeding. No difference was noted in NPY levels measured in the arcuate nucleus and the paraventricular nucleus, in line with the similar amounts of food eaten by all rats at the end of the experiment. CONCLUSION: Increasing plasma leptin concentrations just prior to the end of a period of food restriction reduced subsequent food intake, but did not appear to exert a major influence on the body weight 'set-point', as leptin did not prevent weight regain. The results of the present study suggest that leptin may be of little value in maintaining weight loss in individuals who have lost weight through dieting. Further research is required to understand the role of leptin in the regulation of energy balance. PMID- 12532154 TI - Acute effects of ethanol on feeding behavior and leptin-induced STAT3 phosphorylation in rat hypothalamus. AB - OBJECTIVE: Drinking ethanol stimulates the appetite, producing a positive energy balance. The mechanism by which ethanol regulates the appetite in the central nervous system, however, has not been fully understood. The aim of this study is to investigate the interaction of ethanol with the satiety effect of leptin, a hormone which suppresses the appetite in the hypothalamic region. DESIGN: : Leptin (7.5 micro g) or the same dose of phosphate buffer saline (PBS) was administered into the third ventricle (i.c.v.), 30 min after an intraperitoneal injection (i.p.) of ethanol (0.5 g/kg body weight) or the same dose of PBS. MATERIALS: Adult male Sprague-Dawley rats weighing 290-320 g were used. MEASUREMENTS: Food intake was measured 2, 12 and 24 h after leptin i.c.v. infusion. The tyrosine phosphorylation of signal transducer and activator transcription factor 3 (STAT3) in the hypothalamus was analyzed by Western blotting. RESULTS: The cumulative food intakes in the saline/leptin group (saline i.p.+leptin i.c.v.) were markedly reduced to about 45% of the saline/PBS group (saline i.p.+PBS i.c.v.) at 2, 12 and 24 h time points (P<0.05, 0.001, and 0.005, respectively). As compared with the saline/leptin group, those of the ethanol/leptin group (ethanol i.p.+leptin i.c.v.) were significantly increased to the level seen in the saline/PBS group at 12 and 24 h time points (P<0.05, and P<0.005 vs the saline/leptin group, respectively). Ethanol administration resulted in about a 50% reduction of the leptin-induced STAT3 tyrosine phosphorylation seen in the hypothalamic protein as compared to that of the saline/leptin group. CONCLUSION: These findings suggest that ethanol-induced enhancement of the appetite may, in part, result from leptin resistance transiently caused by ethanol to attenuate the leptin signal transduction. PMID- 12532155 TI - Up-regulation of muscle uncoupling protein 3 gene expression in mice following high fat diet, dietary vitamin A supplementation and acute retinoic acid treatment. AB - OBJECTIVE: To analyse the impact of vitamin A supplementation of both a normal fat (NF) diet and a high fat (HF) diet and of acute retinoic acid (RA)-treatment on the expression of uncoupling protein 3 (UCP3) in mice. DESIGN: C57BL/6J mice were fed for 18 weeks a NF or a HF diet (10 and 45 energy% as fat, respectively), both with the normal vitamin A content or an excess vitamin A (8 mg and 320 mg retinyl palmitate/kg diet, respectively). Body weight and energy intake were recorded periodically. UCP3 mRNA and UCP3 protein levels in skeletal muscle (soleus/gastrocnemius) were analysed, as well as UCP1, UCP2 and UCP3 mRNA levels in interscapular brown adipose tissue (BAT), and UCP2 mRNA, UCP2 protein and leptin mRNA levels in white adipose tissue (WAT) depots. The effect of acute RA treatment (100 mg/kg/day, 4 days) on UCP3 mRNA levels in skeletal muscle and BAT of NMRI mice was also assessed. RESULTS: Vitamin A supplementation of a NF diet led to increased levels of UCP3 mRNA and UCP3 protein in muscle, UCP1 mRNA in BAT, and UCP2 mRNA in inguinal WAT, but had no impact on body weight or adiposity of B6 mice. HF diet promoted obesity and increased levels of UCP3 mRNA and UCP3 protein in skeletal muscle, and of the mRNAs for all three UCPs in BAT. Supplementing the HF diet with vitamin A had little effect on the final obesity reached and did not lead to further increases of muscle UCP3 mRNA nor BAT UCP1 mRNA over the levels achieved with the non-supplemented HF diet. Adipose leptin mRNA levels were down regulated after vitamin A supplementation, independently of the fat content of the diet. Up-regulation of muscle, but not BAT, UCP3 mRNA levels was also found after acute RA-treatment in NMRI mice. CONCLUSION: The results provide evidence of a stimulatory effect of retinoids on muscle UCP3 expression in vivo, and a differential retinoid-regulation of the UCP3 gene in muscle and BAT. PMID- 12532157 TI - Androgen receptor gene repeats and indices of obesity in older adults. AB - OBJECTIVE: To examine the relationship between GGN and CAG trinucleotide repeat polymorphisms in the androgen receptor (AR) gene and indices of obesity in older Caucasian adults. DESIGN: A cross-sectional study. SUBJECTS: Ninety-nine healthy men (age 51-93 y) and 113 healthy postmenopausal women (age 51-92 y). MEASUREMENTS: Genotyping the GGN and CAG repeats of the AR gene, and measuring body weight, height and waist and hip circumferences. Waist was measured at the umbilicus (wstumb), iliac crest (wstili), and mid-way between the iliac crest and lowest rib (wstwst). Waist-to-hip ratios (WHRUMB, WHRILI and WHRWST) and body mass index (BMI) were calculated. RESULTS: Women who were homozygous for a common GGN (17 or 18) and short CAG (/=40 kg/m(2)) Finns. Gel shifts were used to examine whether a mutation in the putative promoter alters GATA-factor binding. RESULTS: Three common MC3R variants were found: a 17C>A variant, changing Thr6- >Lys in 16%, a 241G>A variant changing Val81-->Ile in 15%, and a -239A>G substitution in the GATA binding site in 21% of the subjects. Four other variants were detected in the 5(') flanking region. Frequencies of the three common variants did not differ between obese and contol subjects. Among the obese, the 17C>A and 241G>A variants were coinherited and associated with increased insulin glucose ratios (P<0.05) and leptin levels (P<0.05). GATA-4 bound efficiently to wild type oligonucleotide, but only weakly to the oligonucleotide with the 239A>G mutation. CONCLUSIONS: MC3R gene variants are common and do not explain human morbid obesity. These variants associated with subtle changes in onset of weight gain, hyperleptinemia and insulin-glucose ratios. The -239A>G mutation abolishes binding of GATA-4 to the MC3R promoter region. PMID- 12532158 TI - Insulin resistance but not visceral adipose tissue is associated with plasminogen activator inhibitor type 1 levels in overweight and obese premenopausal African American women. AB - OBJECTIVE: To compare plasma plasminogen activator inhibitor type 1 (PAI-1) levels and to examine the association of PAI-1 with visceral adiposity and other components of the metabolic syndrome in overweight and obese premenopausal African-American (AA) and Caucasian (CC) women. DESIGN: Cross-sectional study. SUBJECTS: 33 CC and 23 AA healthy, overweight and obese, premenopausal women (age 19-53 y, body mass index 28.1-48.9 kg/m(2)). MEASUREMENTS: Body mass index, sagittal diameter, waist circumference, percentage body fat, visceral and subcutaneous adipose tissue (by anthropometry, magnetic resonance imaging (MRI), and bioelectric impedance techniques), PAI-1, leptin, lipids, glucose, insulin, and insulin resistance (by HOMA IR). RESULTS: AA women had lower triglyceride levels and less visceral adipose tissue (VAT) volume than CC despite similar BMI. PAI-1 levels were not significantly different in the two groups. Insulin resistance was associated with PAI-1 in both groups but only in CC women were VAT, triglyceride, HDL cholesterol and blood pressure related to plasma PAI-1 levels. Multiple regression analysis showed that VAT in CC and insulin resistance in AA were independent predictors of PAI-1. CONCLUSION: VAT is significantly associated with circulating PAI-1 levels in overweight and obese CC but not AA premenopausal women. PMID- 12532159 TI - Tumor necrosis factor alpha and insulin resistance in obese type 2 diabetic patients. AB - The relationship between basal serum tumor necrosis factor alpha (TNFalpha) levels and peripheral tissue (muscle) sensitivity to insulin was examined in 63 subjects with normal glucose tolerance (NGT), 18 subjects with impaired glucose tolerance (IGT), and 123 patients with type 2 diabetes mellitus (T2DM). The BMI was similar in NGT (28.8+/-0.7 kg/m(2)), IGT (31.1+/-1.0), and T2DM (30.0+/-0.4) groups. The fasting serum TNFalpha concentration in T2DM (4.4+/-0.2 pg/ml) was significantly higher than in NGT (3.1+/-0.2) and IGT (3.4+/-0.2; both P<0.05). In T2DM the fasting plasma glucose (FPG=183+/-5 mg/dl) and insulin (FPI=17+/-1 micro U/ml) concentrations were significantly higher than in NGT (FPG=95+/-1; FPI=10+/ 1) and IGT (FPG=100+/-2; FPI=13+/-1; all P<0.01). The rate of total body insulin mediated glucose disposal (Rd; 40 mU/m(2) min euglycemic insulin clamp in combination with (3)H-glucose) was reduced in T2DM (102+/-3 mg/m(2) min) compared with NGT (177+/-10) and IGT (151+/-14; both P<0.01). The serum TNFalpha concentration was inversely correlated with Rd (r=-0.47, P<0.0001) and positively correlated with both FPG (r=0.32, P=0.004) and FPI (r=0.32, P=0.004) in NGT plus IGT. No correlation was observed between serum TNFalpha and Rd (r=-0.02), FPG (r=0.15), or FPI (r=0.15) in T2DM. In stepwise multiple regression analysis using age, sex, BMI, FPG, FPI and serum TNFalpha concentration as independent variables, only BMI and serum TNFalpha concentration were significant and independent predictors of Rd (r(2)=0.29, P<0.0001) in the NGT plus IGT group, while FPG and FPI were significant and independent predictors of Rd (r(2)=0.13, P<0.0001) in T2DM. These results suggest that: (i) an increase in circulating TNFalpha concentration is associated with peripheral insulin resistance and increased plasma glucose and insulin levels prior to the onset of type 2 diabetes; and (ii) the further deterioration in peripheral insulin resistance in T2DM (compared with NGT and IGT) is unrelated to the increase in serum TNFalpha concentration. PMID- 12532160 TI - Medium- versus long-chain triglycerides for 27 days increases fat oxidation and energy expenditure without resulting in changes in body composition in overweight women. AB - OBJECTIVE: To determine the effects of long-term consumption of medium chain (MCT) versus long chain triglycerides (LCT) on energy expenditure (EE), substrate oxidation and body composition. HYPOTHESIS: MCT consumption will not result in greater EE, substrate oxidation, and body weight loss compared with LCT consumption. RESEARCH METHODS AND PROCEDURES: Seventeen healthy obese women participated in this randomized, crossover inpatient trial. Meals were prepared and consumed on site for two periods of 27 days. Diets containing 40% of energy as fat, with treatment fat comprising 75% of the total fat, were designed to supply each subject with their individual weight-maintaining energy needs. The MCT diet contained 67% of treatment fat as MCT oil (49% octanoate, 50% decanoate) whereas the LCT diet contained exclusively beef tallow as treatment fat. Body composition was assessed by magnetic resonance imaging (MRI) on day 1 and 28 of each phase while energy expenditure was measured on day 2 and 27. RESULTS: Changes in total and subcutaneous adipose tissue volumes following consumption of MCT and LCT were not different (-0.61+/-0.38 l vs -0.54+/-0.48 l and -0.58+/-0.35 l vs -0.48+/-0.40 l, respectively). Average EE and fat oxidation were greater (P<0.05) during MCT than LCT consumption (0.95+/-0.019 vs 0.90+/-0.024 kcal/min, respectively, for EE and 0.080+/-0.0026 vs 0.075+/-0.0022 g/min, respectively for fat oxidation). DISCUSSION: These results show that long-term consumption of MCT enhances EE and fat oxidation in obese women, when compared to LCT consumption. The difference in body composition change between MCT and LCT consumption, although not statistically different, was consistent with differences predicted by the shifts in EE. It can be concluded that substitution of MCT for LCT in a targeted energy balance diet may prevent long-term weight gain via increased EE. PMID- 12532161 TI - Predictive value of early weight loss in obesity management with orlistat: an evidence-based assessment of prescribing guidelines. AB - OBJECTIVE: To assess the clinical usefulness of published guidelines for the use of orlistat, by studying whether weight loss >/=2.5 kg during a 4 week dietary lead-in period, and weight losses of >/=5% after 12 weeks and >/=10% after 6 months of drug therapy predict weight loss and risk factor changes after 2 years. DESIGN: A retrospective analysis of pooled data from 2 multicentre, randomised, placebo-controlled clinical trials with similar design. SETTING: Twenty-nine centres throughout Europe. PARTICIPANTS: Two hundred and twenty men and women (BMI 28-43 kg/m(2)) who completed 2 years of treatment. INTERVENTION: After a 4 week hypocaloric diet plus placebo, 2 years of treatment with orlistat 120 mg tid, plus a hypocaloric diet for the first year and a weight maintenance diet in year two. MAIN OUTCOME MEASURES: Weight loss and obesity-related risk factor changes. RESULTS: Weight loss >/=5% body weight after 12 weeks of diet plus orlistat therapy was a good indicator of 2 year weight loss, whereas weight loss of >/=2.5 kg during the 4 week lead-in and >/=10% after 6 months did not add significantly to the prediction of 2 year outcomes. Patients who lost >/=5% of their weight at 12 weeks (n=104, 47.3%) lost significantly more weight after 2 years than others: -11.9% (95% confidence interval (CI) -13.4% to -10.3%) vs 4.7% (-5.7% to -3.7%) (P=0.0001), and had significantly greater reductions in total cholesterol, LDL-cholesterol, triglycerides, glucose, insulin, and blood pressure. Among those who achieved >/=5% weight loss at 12 weeks, the overall health benefits were not significantly greater in patients who went on to lose >/=10% body weight at 6 months compared with those who did not achieve >/=10% weight loss by month 6. CONCLUSIONS: Of the criteria currently suggested for assessing response to orlistat treatment, weight loss of >/=5% at 12 weeks accurately predicts sustained improvements in weight and major risk factors at 2 years, while other suggested criteria are less useful. PMID- 12532162 TI - Bone metabolism and risk of secondary hyperparathyroidism 12 months after gastric banding in obese pre-menopausal women. AB - OBJECTIVE: To evaluate, during the first postoperative year in obese pre menopausal women, the effects of laparoscopic gastric banding on calcium and vitamin D metabolism, the potential modifications of bone mineral content and bone mineral density, and the risk of development of secondary hyperparathyroidism. SUBJECTS: Thirty-one obese pre-menopausal women aged between 25 and 52 y with a mean body mass index (BMI) of 43.6 kg/m(2), scheduled for gastric banding were included. Patients with renal, hepatic, metabolic and bone disease were excluded. METHODS: Body composition and bone mineral density (BMD) were measured at baseline, 6 and 12 months after gastric banding using dual energy X-ray absorptiometry. Serum calcium, phosphate, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase, bilirubin, urea, creatinine, uric acid, proteins, parathormone, vitamin D(3), IGF 1, IGF-BP3 and telopeptide, as well as urinary telopeptide, were measured at baseline and 1, 3, 6, 9 and 12 months after surgery. RESULTS: After 1 y vitamin D3 remained stable and PTH decreased by 12%, but the difference was not significant. Serum telopeptide C increased significantly by 100% (P<0.001). There was an initial drop of the IGF-BP3 during the first 6 months (P<0.05), but the reduction was no longer significant after 1 y. The BMD of cortical bone (femoral neck) decreased significantly and showed a trend of a positive correlation with the increase of telopeptides (P<0.06). The BMD of trabecular bone, at the lumbar spine, increased proportionally to the reduction of hip circumference and of body fat. CONCLUSION: There is no evidence of secondary hyperparathyroidism 1 y after gastric banding. Nevertheless biochemical bone markers show a negative remodelling balance, characterized by an increase of bone resorption. The serum telopeptide seems to be a reliable parameter, not affected by weight loss, to follow up bone turnover after gastroplasty. PMID- 12532163 TI - Gestational weight gain and postpartum behaviors associated with weight change from early pregnancy to 1 y postpartum. AB - OBJECTIVES: (1) To describe the relative importance of gestational weight gain, postpartum exercise, food intake and breastfeeding to weight change from early pregnancy to 1 y postpartum; and (2) to identify subgroups of women at greatest risk for major weight gain surrounding childbearing. DESIGN: A prospective cohort study of women who registered for obstetrical care in a hospital and primary care clinic system serving a 10 county area of upstate New York. SUBJECTS: A total of 540 healthy adult women who gave birth to full-term singleton infants. MEASUREMENTS: Sociodemographic characteristics, exercise, food-related behaviors and breastfeeding were assessed using the medical record and a mailed questionnaire. Body weight was measured at prenatal visits and 1 y postpartum. Weight retained and major weight gain (4.55 kg) at 1 y postpartum were the main outcomes. ANALYSIS: Linear and logistic regression analyses were conducted. RESULTS: Women were on average 1.51+/-5.95 kg heavier at 1 y postpartum than they were in early pregnancy. Nearly 25% of women experienced a major weight gain of 4.55 kg or more at 1 y postpartum. Gestational weight gain, exercise frequency, change in food intake and breastfeeding were each significantly related to postpartum weight retention. With the exception of breastfeeding, all of these factors were also associated with major weight gain. Women under 20 y or over 40 y at delivery, and single women retained significantly more weight. Lower income women with gestational weight gains above the Institute of Medicine (IOM) range retained 3.73 kg more than lower income women who gained within the range. They were also 4.7 times more likely to experience major weight gain with childbearing. The impact of exceeding the IOM gestational weight gain guidelines was three times greater in lower income women than it was in higher income women. CONCLUSION: Gestational weight gain, postpartum exercise frequency, and food intake are significantly associated with weight change from early pregnancy to 1 y postpartum and major weight gain with childbearing. Lower income women who gain more weight in pregnancy than the IOM recommends are at high risk for major weight gain with childbearing. PMID- 12532164 TI - Blood pressure and pulse pressure development in a population sample of women with special reference to basal body mass and distribution of body fat and their changes during 24 years. AB - OBJECTIVE: To study blood pressure and pulse pressure longitudinally and their association with basal and change of body mass index (BMI) and waist to hip ratio (WHR). DESIGN: A prospective population study of 1462 women in Gothenburg, Sweden, aged 38-60 y at baseline, with a longitudinal follow-up of 24 y. OUTCOME MEASURES: Incidence of hypertension, systolic and diastolic blood pressure, and pulse pressure at baseline and after 12 and 24 y of follow-up. RESULTS: Systolic and diastolic blood pressure as well as pulse pressure increased with age and turned down again at high age. BMI and WHR at baseline were each independently associated with baseline systolic and diastolic blood pressure, but only BMI with pulse pressure. However, baseline BMI and WHR were not associated with change of systolic, diastolic or pulse pressure during 12 or 24 y of follow-up. Increase in BMI during the follow-up period was associated with increase in systolic and diastolic blood pressure but not with increase in pulse pressure. There were no such associations with WHR changes which, were either unrelated or in one analysis inversely related with blood pressure changes. When considering incidence of hypertension during the first 12 y of follow-up, BMI and change in BMI were significant predictors, independent of WHR. CONCLUSION: Age, BMI and increments in BMI seem to be strong predictors for hypertension and increased systolic and diastolic blood pressure in women. In contrast, WHR plays a lesser and uncertain role in the development of hypertension in middle-aged women. Changes in BMI seem not to be accompanied by changes in pulse pressure during a long time follow-up. PMID- 12532165 TI - Overweight and obesity in Saudi females of childbearing age. AB - OBJECTIVE: Overweight and obesity are among the most frequently encountered multifactorial disorders in most populations of the world. The aim of this study was to determine the prevalence of overweight and obesity in Saudi females of childbearing age. MATERIALS AND METHODS: The study was approved by the University Scientific Committee. It included a group of 600 randomly recruited healthy females with age ranging from 16-45 y. Four hundred and twenty one were students and 179 were housewives attending outpatient clinics for minor illnesses. Informed consent was obtained. Height and weight were recorded on one occasion. Body mass index (BMI) was calculated. RESULTS: Height, weight and BMI showed normal Gaussian distribution in these females and there was a significant positive correlation between BMI and age of each subject (r=0.505; P/=30 kg/m(2). RESULTS: The prevalence of obesity was 36.8 and 18.1% in rural women and men, respectively, compared with 49.1 and 30.6% in urban women and men, respectively. The mean difference (s.e) in BMI levels was 1.6 (0.52) kg/m(2) between urban and rural women and 0.9 (0.46) kg/m(2) in men. At the household level, the mean energy consumption from 25 selected food items was 13.8 MJ (3310 kcal)/consumption unit/day in the rural community compared to 14.5 MJ (3474 kcal)/consumption unit/day in the urban community (P=0.021). BMI was positively associated with age in both men and women and with urban residence in women. BMI was negatively associated with smoking and physical activity in men and with educational level in women. CONCLUSION: BMI was associated with urban residence in women after adjusting for age, smoking, education, physical activity and nutrition-related variables, suggesting that the differences in the conventional determinants of obesity could not fully explain the difference in the prevalence of obesity between the two communities. Among men, the measured determinants explained the rural-urban differences in BMI. PMID- 12532167 TI - Contemporary revascularization strategies in 2002. AB - This review analyzes important facets of contemporary percutaneous coronary interventions. The optimal strategy of acute myocardial infarction and shock is addressed, as is the role of angioplasty in multivessel coronary disease. Vascular Brachytherapy is essentially the sole available treatment modality for in-stent restenosis. The WRIST trials have provided the foundation for clinical experience and are discussed in detail. Finally, drug-eluting stents may become the next revolution in interventional cardiology and offer the hope of a "cure" for restenosis. PMID- 12532168 TI - Mother-to-child transmission of hepatitis C virus infection. AB - The prevalence of hepatitis C virus (HCV) infection among pregnant women in Europe is generally below 2%. Although women with a history of parenteral exposures or injecting drug use are at an increased risk of infection, a substantial proportion of infected women do not report any risk factors. Targeted screening is thus not recommended. The risk of mother-to-child, or vertical, transmission of HCV is about 5% overall but can be as high as 15%, depending on maternal HIV infection status and HCV RNA viral load. Larger studies are needed to confirm or refute the potential protective effect of elective cesarean section delivery. However, for HCV-positive women who are co-infected with HIV, elective cesarean section delivery is associated with a reduced risk of vertical transmission of HIV as well as HCV. The risk of postnatal transmission through breastfeeding cannot be excluded but is likely to be low for most HCV-infected women. The long-term natural history of vertically acquired HCV needs further elucidation and the efficacy of potential therapies for infected children need to be evaluated in randomized controlled trials. PMID- 12532169 TI - Statins in acute coronary syndromes. AB - The development of 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase inhibitors (statins) has been a very significant development in the management of coronary artery disease. Large prospective clinical trials have provided unequivocal evidence that cholesterol lowering therapy with statins reduces all cause mortality in patients with coronary artery disease. There is now accumulating data indicating that statin treatment should be initiated early after an acute coronary syndrome. This body of evidence is based on large databases in which investigators compared outcomes among patients taking statins with those patients who were not prescribed cholesterol lowering therapy. Prospective, randomized, clinical trials also indicate that early statin therapy reduces recurrent ischemia. Finally, studies examining long-term compliance with statin therapy suggest increased adherence to therapy when statins are prescribed during the initial hospitalization for an acute coronary syndrome. In tandem with these clinical observations, there is a large body of scientific data that highlights many important cellular and molecular mechanisms through which statins may confer early benefit. These effects involve relatively rapid improvement in endothelial function, antiischemic, antithrombotic and antiinflammatory actions of statins. PMID- 12532170 TI - Molecular biological and genetic approaches to the evaluation of inherited electrophysiologic disorders. AB - Although previously thought of as sporadic errors in development, some arrhythmias are now established to have a hereditary basis. The genetic and molecular bases of inherited cardiac arrhythmias are being recognized with increased frequency. Environmental and somatic influences may complicate classical identification of the patterns of inheritance. In several examples of inherited heart disease with a distinct molecular and genetic foundation, multiple gene defects appear to be responsible for similar phenotypes. In addition, specific genotypes may lead to differing severity of phenotype and differential prognostic risks. There is also variability in phenotypic expression, even among identical genotypes within a family, complicating risk stratification. Improved understanding of rare familial arrhythmia disorders may increase our overall comprehension of the pathophysiology of rhythm disturbances in general. The inherited arrhythmia disorders can be classified under three broad headings -- those familial arrhythmias that originate above the ventricles, familial arrhythmias originating within the ventricles and familial disorders affecting impulse propagation and cardiac conduction. Familial occurrences of tachyarrhythmias and bradyarrhythmias have been reported for each of these types of disorders. Recent discoveries of genetic etiologies have shed new light on the mechanisms underlying arrhythmogenesis and molecular genetics of cardiac electrophysiology. PMID- 12532171 TI - Chronic cerebral hypoperfusion-related neuropathologic changes and compromised cognitive status: window of treatment. AB - Neurodegenerative disorders, and dementia in particular, have been shown to have a cerebrovascular pathogenic component often in the form of reduced cerebral blood flow. The debate whether such a reduced brain perfusion is a primary trigger or a secondary symptom in the neuropathological progression of dementia has not been conclusively decided yet. However, compelling experimental evidence has been collected to demonstrate an initiating role of reduced cerebral blood flow in neurodegenerative processes. Along these lines, experimental cerebral hypoperfusion in rodents was shown to impair spatial learning and to generate neuronal damage and associated gliosis in sensitive brain regions like the hippocampus and frontoparietal cortex. Since suboptimal cerebral blood supply was thus identified as a potential trigger of cognitive decline, the improvement of cerebral blood flow in cognitive disorders has emerged as an alternative treatment to moderate the symptoms and to delay the onset of advanced dementia. Various drugs, such as cholinergic compounds, hemorheologic agents and vascular smooth muscle relaxants, have already been tested in some instances for their efficacy to increase brain perfusion. In this respect, both clinical and preclinical trials delivered positive data. Furthermore, not only the treatment but also the prevention of the development of cognitive deficiency can target the cerebrovascular system. For this purpose, long-chain polyunsaturated fatty acids derived from fish oil (also known as n-3 PUFAs) have been considered as dietary supplements. These fatty acids appeared particularly effective in the prevention of hypertension-associated vascular pathology. The present review provides an overview of the actions of these compounds focusing on cerebral blood flow, neurodegeneration and cognitive decline. PMID- 12532172 TI - Three-dimensional visualization and analysis in prostate cancer. AB - Current and emerging three- and four-dimensional medical imaging modalities, along with development of efficient 3-D computer rendering and modeling of multidimensional volume image data and image-guided navigation, are significantly advancing our capabilities for improved and minimally invasive diagnosis and treatment of prostate cancer, obviating the need for exploratory surgery, physical dissection, blind biopsies and mental reconstruction of anatomy and pathology. Currently, both diagnostic and therapeutic procedures require x-ray fluoroscopy, transrectal ultrasound, CT and/or MRI for assessing the condition of the prostate and/or the outcome of any therapeutic procedure. New imaging approaches based on three-dimensional ultrasound transducers placed on catheters for easy insertion into the urethra are demonstrating significant promise for improved diagnosis and treatment of prostate disease. Microwave thermal ablation shows promise for reduction of prostate size and tumor volume, and preliminary data from cryosurgery suggests improvements in tumor reduction and/or management while minimizing the risk of serious complications. Prostate brachytherapy is becoming a more popular and effective alternative to surgery. All of these methods, either independently or combined through image fusion, are providing an exciting and rapid evolution in capabilities for visualizing the prostate and its anatomic environment, extending from physical to functional forms and from macro to micro orders of scale. Traversing the scale distances between these imaged objects within the prostate and its environs will be made automatic and instantaneous in the near future with the expected advances in miniaturization of powerful computing and electronic sensing elements. Imaging devices will continue to improve in resolution, speed and affordability and will be deployed harmlessly within the body, as well as outside of it. Diagnosis and therapy of prostate disease will become fully noninvasive and synchronous. PMID- 12532173 TI - Genetic basis of systemic lupus erythematosus. AB - Systemic lupus erythematosus, a complex multigenic disease, is characterized by a wide variety of clinical manifestations, as a result of the contribution of different genes or different combinations of genes. Recent advances in molecular genetics has led to a better understanding of the genes predisposing to systemic lupus erythematosus in both humans and laboratory animal models. Identification of susceptibility genes provides key insights into the pathogenesis of systemic lupus erythematosus, making new prophylactic and therapeutic approaches feasible. PMID- 12532174 TI - Fondaparinux sodium. AB - Fondaparinux (Org-31540 / SR-90107A) is a new drug chemically synthesized for treatment and prophylaxis of thromboembolic disease. Fondaparinux is a selective inhibitor of activated factor X. Its structure is the copy of the heparin pentasaccharide sequence, the shortest chain required for antithrombin inhibition of activated factor X without antithrombin action. Fondaparinux has no effect on coagulation tests and does not bind to platelet factor 4 or promote heparin induced thrombocytopenia. Fondaparinux inhibits thrombin generation and the growth of thrombi in in vitro and in vivo models. Phase I trials have shown a 100% bioavailability after subcutaneous (s.c.) administration, a rapid onset of action and an approximate half-life of 13.5 h. Fondaparinux is cleared as an active substance by the kidneys. In elderly patients, renal clearance is reduced and the half-life is longer. The phase II Pentathlon trial demonstrated significant dose-dependent reductions in the frequency of venous thromboembolism in total hip-replacement patients and the optimal dose was determined to be 2.5 mg s.c./24 h. Four phase III trials have evaluated fondaparinux starting 6 hours after surgery compared with enoxaparin for prevention of venous thromboembolism following orthopedic surgery in 7,344 patients. The risk of thrombosis was reduced by 50% with fondaparinux and no differences were observed in death or severe bleeding. In a phase II trial, similar efficacy and incidence of major bleeding were seen with fondaparinux s.c. compared with dalteparin s.c. in the treatment of deep venous thrombosis. In patients with acute myocardial infarction, the efficacy of fondaparinux during fibrinolytic therapy was assessed in 326 patients who had acute coronary syndromes of less than a 6 hour duration, showing a slight but statistically not significant advantage for fondaparinux over unfractionated heparin in the coronary angiographies. There is currently no antidote for fondaparinux. PMID- 12532175 TI - Ertapenem. A review of its microbiologic, pharmacokinetic and clinical aspects. AB - Ertapenem sodium (trade name Invanz, also designated as MK-0826, MK-826 and L 749345), manufactured by Merck & Co., Inc. is a structurally unique parenteral 1 beta-methyl carbapenem. It has a broad spectrum of antimicrobial activity, including common community-acquired Gram-positive and Gram-negative aerobic and anaerobic pathogens, and restricted activity against nosocomial pathogens such as Pseudomonas aeruginosa, Acinetobacter species, methicillin-resistant staphylococci and enterococci. Ertapenem demonstrates excellent activity against cephalosporin-resistant enteric organisms producing extended spectrum beta lactamases (ESBLs) or AmpC beta-lactamases and excellent activity against penicillin-resistant Streptococcus pneumoniae. Its high level of protein binding and serum half-life of 4 h allows it to be dosed once daily. Ertapenem may be administered intravenously or intramuscularly and has an excellent adverse reaction and tolerability profile. Invanz was approved by the United States Food and Drug Administration in November 2001 for the treatment of adult patients with moderate to severe infections caused by designated strains of susceptible microorganisms. The infections include complicated intraabdominal infections, complicated skin and skin structure infections, community-acquired pneumonia, complicated urinary tract infections including pyelonephritis and acute pelvic infections including postpartum endomyometritis, septic abortion and post surgical gynecologic infections. Invanz has also been approved in Mexico, Brazil and New Zealand. PMID- 12532176 TI - Exploring MEDLINE abstracts with XplorMed. AB - XplorMed is a publicly available web tool conceived to make life easier for MEDLINE(c) users looking for scientific information. Searching scientific literature is an information retrieval problem. Abstracts that are of possible interest to the user are usually selected by a keyword search followed by manual screening, which often results in the retrieval of a large number of abstracts. Interesting references can be buried among irrelevant ones because of nonspecific queries. XplorMed is intended to extract dependency relations between the words of the abstracts. These relations can be filtered and arranged to deduce different subjects in the query and offer a condensed view of the abstract, allowing users to select texts of interest without having to read them all. XplorMed is available http://www.bork. embl-heidelberg.de/xplormed. PMID- 12532177 TI - The immunogenetics and pathogenesis of gastric cancer. Highlights of the First Sino-European Workshop on the Immunogenetics and Pathogenesis of Gastric Cancer. AB - Clinical scientists from eight European countries and China gathered in the ancient Chinese capital of Xi'an on April 26-28, 2001 to discuss collaboration on a modern approach to gastric cancer prevention. Participants at the First Sino European Workshop on Immunogenetics and Pathogenesis of Gastric Cancer presented their most up-to-date research results on topics ranging from epidemiology and immune mechanisms to Helicobacter pylori and vaccine development. Researchers then formed groups with their Chinese or European counterparts to plan future research endeavors which will benefit Chinese and European populations alike. After 3 years of organization between the Institute of Digestive Diseases of the Fourth Medical University in Xi'an, China and the Laboratory of Immunogenetics, VU University Medical Center in Amsterdam, the first workshop came into being under the joint sponsorship of the Commission of the European Union, National Natural Science Foundation of China and the Institute of Digestive Diseases, Xi'an, China. As gastric cancer is the most prevalent malignant tumor in China, the workshop was of special significance to the Chinese researchers and to the Chinese population in general. During the workshop, presentations on the epidemiology of gastric cancer showed that this disease is in fact common the world over: it is the second most common cancer next to lung cancer and about 1 million new cases were diagnosed in 2000. Three-quarters of the cases of gastric cancer occur in Asia, and approximately 80% of these cases are in China and Japan. Genetic factors and environmental factors such as diet and H. pylori infection play a role in gastric carcinogenesis. As a recognized cause of gastric cancer, H. pylori was the subject of various presentations ranging from immunological studies, molecular analysis of strains and pathogenesis to vaccine development. Specific areas of discussion included bacterial-epithelial interactions in H. pylori infection, epidemiology in China, global distribution of vacA and cagA genotypes, new evidence for host factors, nonsteroidal antiinflammatory drugs and H. pylori as independent risk factor for gastric cancer, new diagnostic techniques for H. pylori using serum levels of pepsinogen I, and autoimmune processes in corpus atrophy. Vaccine development using a variety of strategies against H. pylori was the subject of an entire session of talks. Oral immunization with urease with Escherichia coli heat labile enterotoxin was shown to be safe and immunogenic in humans as a mucosal adjuvant. Results of a study using attenuated Salmonella typhimurium as a vehicle for DNA mediated immunization in mice were also presented. A final presentation discussed an ongoing trial comparing strain variability in the vacA and cagA gene sequences and disease expression between H. pylori infection in Europe and China. Researchers also discussed the role of IL1 gene family and TNF gene polymorphisms in gastric pathology and various immune mechanisms involved in gastric cancer, such as down-regulation of NF kappa B, IL-1 and IL-1RA, cyclooxygenase signalling, and identification of MGAg antibodies. An interactive discussion followed each presentation and ideas and suggestions were provided. According to specialty, the presenters were then assigned to groups of four or five to make plans for joint research projects. A number of international and Chinese observers were present, including representatives from the European Commission, the World Health Organization and the Chinese National Center for Biotechnology Development, and offered input on the financial feasibility of such projects. PMID- 12532178 TI - Projections for insulin treatment for diabetics. AB - The evolution of insulin treatment of diabetes has dramatically changed the natural course of this disease. Modern recombinant DNA technology has brought about many new insulin analogues with improved pharmacokinetics, resulting in better glycemic control. In addition, improved insulin delivery systems, such as insulin pumps and pens, have been introduced to provide convenience and to enhance patient compliance. Efforts are currently being devoted to developing noninvasive insulin formulations, such as oral and pulmonary insulin. A number of products are at different stages of clinical trials. Meanwhile, the quest for a permanent cure for diabetes continues. The frontier of diabetes research has gone through a period of substantial expansion, with the emergence of new areas that include gene therapy, islet cell transplantation and diabetic vaccine. Technological breakthroughs, such as recombinant DNA, nanotechnology, microarray aided genomics and proteomics, will provide more profound insights into the pathogenesis, and the immunological and biological basis of diabetes. Our growing knowledge in these areas will ultimately contribute to the discovery of preventive methods against or a cure for this disease. PMID- 12532179 TI - The pathological cascade of Alzheimer's disease: the role of inflammation and its therapeutic implications. AB - Alzheimer's disease is a chronic neurodegenerative disease causing progressive impairment of memory and other cognitive functions. A number of sequential events are suggested to be associated with different pathological aspects observed in Alzheimer's disease, the so-called amyloid cascade hypothesis. Mismetabolism of the beta-amyloid precursor protein, as a result of mutations in the amyloid precursor protein gene or as results of impaired cleavage, leads to the formation of nonfibrillar and fibrillar amyloid-beta deposits. Glial cells are attracted to and activated by these amyloid-beta deposits. After activation, these cells secrete inflammatory mediators and reactive oxygen species, which can aggravate the aggregation of amyloid-beta. Some of the products released by activated glial cells, as well as amyloid-beta itself, can induce or promote neurodegeneration. Several mechanisms, such as mitotic reentry, apoptosis and cytoskeletal changes are suggested to be involved in neuronal loss. This review will outline several pathological mechanisms in Alzheimer's disease as well as some means of therapeutic intervention following the amyloid cascade hypothesis. PMID- 12532180 TI - Therapeutic options for Parkinson's disease. AB - Parkinson's disease is a multifactorial disease. In early-onset cases, genetic predisposition may play a role, but typical Parkinson's disease is most likely determined by genetic and environmental factors. Since age is a consistent risk factor, an age-dependent cumulative insult mechanism may be responsible for the selective degeneration of nigrostriatal neurons. Although there is no cure, several types of treatments are currently available to substantially reduce the clinical symptoms of Parkinson's disease. An understanding of the age-dependent complex pattern of genetic variation (SNP genotyping) together with the genetic expression profile (using chip technology) of the degenerating neurons and the effect of environmental factors is a prerequisite to deciphering the mechanism of pathogenesis. This understanding may lead to predicting drug response and ultimately to developing preventive medicine at the individual level in the future. PMID- 12532181 TI - Topical tacrolimus Protopic. AB - Topical tacrolimus is the first topical immunomodulator of its kind, and its mechanism of action, pharmacokinetics, metabolism and efficacy will be discussed. Multiple studies have supported the safety and efficacy of topical tacrolimus in the treatment of atopic dermatitis in both adults and children. Studies have also suggested that topical tacrolimus may be effective in the treatment of acute contact dermatitis. Although oral administration of tacrolimus proved to be efficacious in psoriasis, results with topical tacrolimus have been disappointing in some studies. Additional reports of success in the use of topical tacrolimus in the treatment of pyoderma gangrenosum show promise, although there is a paucity of randomized, placebo-controlled trials examining its use. It has also been suggested that topical tacrolimus is useful in the treatment of alopecia areata, erosive lichen planus, isolated ichthyosis linearis circumflexa, steroid induced rosacea, and in the prevention of allograft rejection, however more clinical trials need to be performed. PMID- 12532182 TI - Pathological mechanisms and disease progression of multiple sclerosis: therapeutic implications. PMID- 12532183 TI - New therapies for the treatment of congestive heart failure. AB - Heart failure is characterized by sodium and fluid retention, sympathetic overactivity, parasympathetic withdrawal, vasoconstrictor activation and cytokine elevation. New therapies for heart failure attempt to control neurohormonal activation and limit progressive left ventricular dysfunction. Nesiritide (human B-type natriuretic peptide) is a recently approved new vasodilator that has been given to almost 1,000 patients in numerous clinical investigations; it belongs to a new class of heart failure drugs known as natriuretic peptides. Nesiritide decreases pulmonary capillary wedge pressure, systemic vascular resistance, mean right atrial pressure and pulmonary artery pressure, while improving cardiac index, stroke volume and heart failure symptoms. Many endothelin receptor antagonists are in various stages of development. Early clinical studies have demonstrated beneficial cardiovascular hemodynamic effects. Other new drugs for heart failure also include calcium sensitizers, neutral endopeptidase and vasopeptidase inhibitors, aldosterone receptor antagonists, vasopressin antagonists and cytokine inhibitors. All are being actively investigated and many show significant promise as beneficial therapies in the treatment of heart failure. PMID- 12532184 TI - Neuroprotective effect of flupirtine in prion disease. AB - Apoptotic neuronal cell death is a hallmark of prion diseases. The apoptotic process in neuronal cells is thought to be caused by the scrapie prion protein, PrPSc, and can be experimentally induced by its peptide fragment, PrP106-126. This process is a target for potential drugs to combat prion disease or to ameliorate its symptoms. Flupirtine (Katadolon), a pyridine derivative that is in clinical use as a nonopioid analgesic, has a potent cytoprotective effect, at concentrations above 1 microg/mL, on neuronal cells treated with PrP(Sc) or PrP106-126. This drug acts as an N-methyl-D-aspartate (NMDA) antagonist, but does not bind to NMDA receptors. Flupirtine normalizes the level of intracellular glutathione and increases the expression of the antiapoptotic Bcl-2 protein in neuronal cells exposed to prion protein. In view of its favorable pharmacokinetic profile, flupirtine is the first drug to be considered as a potential treatment for Creutzfeldt-Jakob disease, the human form of prion diseases. Clinical trials are underway. PMID- 12532185 TI - Medical backgrounder on varicocele. AB - Varicocele is an extremely common entity, present in 15% of the male population. Varicoceles are found in approximately 35% of men with primary infertility but 75%-81% of men with secondary infertility. Mounting evidence clearly demonstrates that varicocele causes progressive duration-dependent injury to the testis. Larger varicoceles appear to cause more damage than small varicoceles and, conversely, repair of large varicoceles results in greater improvement of semen quality. Varicocelectomy can halt the progressive duration-dependent decline in semen quality found in men with varicoceles. The earlier the age at which varicocele is repaired, the more likely is recovery of spermatogenic function. Varicocelectomy can also improve Leydig cell function resulting in increased testosterone levels. The most common complications after varicocelectomy are hydrocele formation, testicular artery injury and varicocele persistence or recurrence. The incidence of these complications can be reduced by employing microsurgical techniques, with inguinal or subinguinal operations, and exposure of the external spermatic and scrotal veins. Employment of these advanced techniques of varicocelectomy provide a safe, effective approach to elimination of varicocele, preservation of testicular function and, in a substantial number of men, an increase in semen quality and likelihood of pregnancy. PMID- 12532186 TI - Ramosetron, a 5-HT3 receptor antagonist for the control of nausea and vomiting. AB - Ramosetron is a selective serotonin 5-HT3 receptor antagonist with an affinity higher than that of the previously available drugs ondansetron, granisetron and tropisetron. Ramosetron was shown in pharmacological assays to inhibit activities mediated by 5-HT3 receptors, such as emesis caused by cisplatin. In clinical trials, ramostreon was at least as effective as the reference "setrons" against nausea and vomiting induced by chemotherapy or surgical interventions, but the efficacy was well maintained during a 48-h period, so that efficacy was significantly higher with ramosetron in terms of nausea and vomiting 6-48 h after treatment. Hence, ramosetron represents a good alternative over previously available drugs for the prophylaxis and treatment of chemotherapy-induced and postoperative nausea and vomiting. Data in children also support the efficacy of ramosetron, which is well tolerated by both child and adult patients. PMID- 12532187 TI - The role of osteoclastic activity in prostate cancer skeletal metastases. AB - Metastasis of prostate cancer to bone is a common complication of progressive prostate cancer. Skeletal metastases are often associated with severe pain and thus demand therapeutic interventions. Although often characterized as osteoblastic, prostate cancer skeletal metastases usually have an underlying osteoclastic component. Advances in osteoclast biology and pathophysiology have led toward defining putative therapeutic targets to attack tumor-induced osteolysis. Several factors have been found to be important in tumor-induced promotion of osteoclast activity. One key factor is the protein receptor activator of nuclear factor-kappa B ligand (RANKL), which is required to induce osteoclastogenesis. RANKL is produced by prostate cancer bone metastases, enabling these metastases to induce osteolysis through osteoclast activation. Another factor, osteoprotegerin, is a soluble decoy receptor for RANKL and inhibits RANKL-induced osteoclastogenesis. Osteoprotegerin has been shown in murine models to inhibit tumor-induced osteolysis. In addition to RANKL, parathyroid hormone-related protein and interleukin-6 are produced by prostate cancer cells and can promote osteoclastogenesis. Finally, matrix metalloproteinases (MMPs) are secreted by prostate cancer cells and promote osteolysis primarily through degradation of the nonmineralized bone matrix. MMP inhibitors have been shown to diminish tumor establishment in bone in murine models. Thus, many factors derived from prostate cancer metastases can promote osteolysis, and these factors may serve as therapeutic targets. The importance of osteoclasts in the establishment and progression of skeletal metastases has led to clinical evaluation of therapeutic agents to target them for slowing metastatic progression. Bisphosphonates are a class of compounds that decrease osteoclast life span by promoting their apoptosis. The bisphosphonate pamidronate has proven clinical efficacy for relieving bone pain associated with breast cancer metastases and has a promising outlook for prostate cancer metastases. Another bisphosphonate, zoledronic acid, appears to directly target prostate cancer cells in addition to diminishing osteoclast activity at the metastatic site. In addition to bisphosphonates, other novel therapies based on studies that delineate mechanisms of skeletal metastases establishment and progression will be developed in the near future. PMID- 12532188 TI - Bone marrow transplantation: a new strategy for intractable diseases. AB - Bone marrow transplantation is becoming a powerful strategy for the treatment of hematologic disorders (leukemia, aplastic anemia, etc.), congenital immunodeficiencies, metabolic disorders and also autoimmune diseases. Using various animal models for autoimmune diseases, we have previously found that allogeneic (not autologous) bone marrow transplantation can be used to treat autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, immune thrombocytic purpura, insulin-dependent diabetes mellitus, chronic glomerulonephritis and certain types of non-insulin-dependent diabetes mellitus. In contrast, we have found that the transplantation of T-cell-depleted bone marrow cells or partially purified hemopoietic stem cells from autoimmune-prone mice to normal mice leads to the induction of autoimmune diseases in the recipients. These findings have recently been confirmed even in humans; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis and Crohn's disease were resolved after allogeneic bone marrow transplantation. However, there have recently been reports on the rapid recurrence or persistence of autoimmune diseases after autologous bone marrow transplantation. Conversely, the adoptive transfer of autoimmune diseases such as myasthenia gravis, insulin-dependent diabetes mellitus and Graves' disease by allogeneic bone marrow transplantation from donors to recipients has been reported. Owing to these findings, we have proposed that autoimmune diseases are "stem cell disorders." We have thus succeeded in treating autoimmune diseases in various autoimmune-prone mice, except MRL/lpr mice, by conventional bone marrow transplantation. The MRL/lpr mouse itself is radiosensitive (<8.5 Gy), while the abnormal hemopoietic stem cells of the MRL/lpr mouse are radioresistant (>8.5 Gy); conventional bone marrow transplantation (8.5 Gy plus bone marrow transplantation) has a transient effect on autoimmune diseases, which recur three months after the bone marrow transplantation. However, bone marrow transplantation plus bone grafts (to recruit donor stromal cells) completely prevents the recurrence of autoimmune diseases in MRL/lpr mice. Donor-derived stromal cells (including mesenchymal stem cells) thus seem to play a crucial role in successful allogeneic bone marrow transplantation, since there is a major histocompatibility complex restriction between hemopoietic stem cells and stromal cells. We have, however, found that the combination of bone marrow transplantation plus bone grafts has no effect on the treatment of autoimmune diseases in MRL/lpr mice, since MRL/lpr mice become more radiosensitive after the onset of lupus nephritis due to the development of uremic enterocolitis. To reduce the cytotoxic effect of radiation on the intestine, we carried out fractionated irradiation and devised a new strategy. We injected allogeneic whole bone marrow cells (including a small number [<3%] of T cells, hemopoietic stem cells and stromal cells) from donors directly into the intra-bone marrow of recipients so that donor-derived hemopoietic cells including stromal cells could effectively accumulate in the bone marrow. All the MRL/lpr mice survived more than one year (>60 weeks after birth) without the recurrence of autoimmune diseases, and immunological functions were completely restored even when the radiation dose was reduced to 5 Gy x 2. These findings suggest that intra-bone marrow injection-bone marrow transplantation can be used to treat intractable autoimmune diseases under reduced radiation doses without using any immunosuppressants.Intra-bone marrow injection-bone marrow transplantation seems to be the best strategy for allogeneic bone marrow transplantation: 1) no graft versus-host disease develops even if T cells are not depleted from the bone marrow; 2) no graft failure occurs even if the dose of radiation as the conditioning for bone marrow transplantation is reduced to 5 Gy x 2; 3) hemopoietic recovery is rapid; and 4) T-cell functions are completely restored even in donor-recipient combinations across the major histocompatibility complex barriers. Using cynomolgus monkeys, we have recently established a new method (the "perfusion method") for collecting bone marrow cells from the long bones (femur, humerus, etc.) without peripheral blood contamination. This method has various advantages: 1) no graft-versus-host disease develops even in cynomolgus monkeys, since the percentage of T cells in the bone marrow cells collected is less than 3%; 2) a large number of bone marrow cells can be collected quickly and safely; and 3) the bone marrow cells collected contain stromal cells including mesenchymal stem cells. We therefore believe that this method (intra-bone marrow injection-bone marrow transplantation in conjunction with the perfusion method) will become a powerful new strategy for not only allogeneic bone marrow transplantation but also organ transplantation in conjunction with bone marrow transplantation. Furthermore, this method could become a valuable strategy in regeneration therapy for injured organs and tissues (myocardial infarction, cerebral infarction, Alzheimer's disease, etc.), since it can efficiently reconstitute the recipient with both donor-derived hemopoietic stem cells and mesenchymal stem cells. PMID- 12532189 TI - Matrices for tissue-engineered skin. AB - Academic, clinical and industrial efforts are increasingly being directed toward the use of molecular- and cell-based therapies for diagnosis and treatment of a great number and broad variety of pathologies and injuries. Hence, tissue engineering is, next to genetic engineering, widely heralded as the healthcare technology heir of the revolutionary advances in life sciences. In a cost controlled healthcare environment, only those technologies capable of providing a major enhancement to quality of life and a reduction in expenditure will be driven forward. Skin tissue engineering concepts based on the application of a scaffold/cell construct represent a treatment concept with the clear potential to meet this criterion. Several important issues concerning the research and product strategy (e.g., choice of matrix of natural or synthetic polymer origin and the use of autogenic versus allogenic cells) remain to be fully resolved. However, there is little doubt that wound regeneration via modern tissue engineering strategies will present significant therapeutic benefits when compared with existing treatments. This paper reviews the biography and future research directions of matrices used in skin tissue engineering. PMID- 12532191 TI - Urological oncology in Europe. Research highlights from the XVIIth congress of the European Association of Urology, Birmingham, UK, February 23-26, 2002. PMID- 12532190 TI - New directions in pain management. AB - Pain medicine is one of the most rapidly developing medical specialties of today. While there are many modalities that can be used in managing the patient in pain, drug treatment remains, for the most part, the cornerstone of treatment. Opioids retain their position as the foundation of most analgesic strategies, although they tend to be used nowadays in combination with adjuvant analgesics such as paracetamol and nonsteroidal antiinflammatory drugs. The range of available opioids has also been expanded with drugs such as hydromorphone and oxycodone, originally developed almost a century ago. This expanded choice has resulted in the concept of opioid rotation in chronic pain states, an approach that is aimed at maintaining pain control while minimizing adverse effects. Nonsteroidal antiinflammatory drugs continue to play an important role, especially as adjuvants, and the development of drugs such as celecoxib and refecoxib, highly specific for the inhibition of cyclooxygenase 2 pathway has been a further advance. The treatment of neuropathic pain continues to be a challenge to the clinician. While this has traditionally been treated with drugs from the anticonvulsant, antiarrhythmic and anti-depressant groups, results from these treatments have often been less than satisfactory. This has led to the development of completely new drug classes that modulate neuronal transmission in pain pathways, some of which are derived from exotic animal sources, such as the conotoxins from the marine snail family and epibatidine from a species of frog. The role of cannabinoids remains controversial. PMID- 12532192 TI - Strategies to identify disease genes. AB - The correlation between genes and disease began in earnest in the early 1900s with the identification of Mendelian-like inheritance of "inborn errors of metabolism." Since then, the ever-broadening field of genetics has been established as one of the most important and groundbreaking branches of science and medicine to date. With the announcement of a "working draft" sequence of the human genome in 2001, the vast array of both genomic and expressed sequence information available in the public databases alone has meant that the concept of hunting for genes is evolving. Nowadays, researchers can substitute many labor intensive hours in the lab for less time searching on the World Wide Web. Specialization within genetics has been continuously providing subsets of the genre such as genomics, pharmacogenetics, chemogenomics, gene therapy, proteomics and functional genomics, all of which are based on the fundamental starting block, the gene. This review aims to summarize both traditional and current strategies for identifying susceptibility and monogenetic disease genes and describes how these strategies have evolved in tune with the ever-expanding wealth of information now available at our fingertips. PMID- 12532193 TI - Micafungin sodium (FK-463). AB - FK-463 (micafungin) represents the latest development candidate in a novel chemical class of echinocandin lipopeptide antifungal compounds. This agent has potent in vitro and experimental in vivo activity against a variety of pathogenic Candida species (yeasts) and Aspergillus fumigatus (filamentous fungus). This compound has favorable pharmacokinetics and a unique mode of action that makes it active against fungal isolates resistant to established antifungal agents, particularly the triazole agent fluconazole. Single- and multiple-dose phase I studies in normal human volunteers and phase II clinical trials in patients have been completed, with the compound being generally well tolerated and efficacious against infections caused by Candida and Aspergillus species. Published information on the in vitro and experimental in vivo activity, experimental and human pharmacokinetics, and clinical trial data of this new antifungal, echinocandin-like lipopeptide are summarized in this monograph. PMID- 12532194 TI - Systemic lupus erythematosus. AB - Systemic lupus erythematosus is a pleomorphic, autoimmune disorder of unknown cause that can afflict all parts of the body. New insights into its pathogenesis, the employment of preventive measures and combinations of immunosuppressive therapy, as well as the introduction of biologics, are beginning to alter the prognosis and improve the quality of life for individuals with this serious disease. PMID- 12532195 TI - Herbal remedies for asthma treatment: between myth and reality. AB - Asthma is one of the most common chronic diseases worldwide. To treat this widespread disease there is a high prevalence of usage of herbal medicine. The use of plants is as old as humankind and it has been steadily increasing over the past 10 years. Plant-based remedies are now one of the most popular complementary treatments. Herbal supplements are receiving increasing exposure through media, including the Internet, in lay journals and more recently in the scientific press. Interest in herbal medicine has been facilitated by multiple factors, including the perception that pharmaceutical medications are expensive, overprescribed and may often be dangerous. Alternatively, herbal medicine is often perceived as being "natural" and therefore is considered safe. However, the scientific literature supporting the efficacy of herbal therapies is incomplete. There are few well-controlled studies that support the efficacy of herbal remedies in the treatment and clinical improvement of patients with asthma. Available scientific evidence has not yet confirmed the validity of their popular role in the treatment of asthma. The present review evaluates herbs and their efficacy in asthma to provide a balanced and objective view for the reader seeking information on herbal therapy PMID- 12532196 TI - Assessing and improving doctors' understanding of Helicobacter pylori infection. PMID- 12532197 TI - Serum levels of soluble intercellular-1 and vascular cell-1 adhesion molecules in chronic hepatitis C and the influence of interferon-alpha + ribavirin therapy. AB - In chronic hepatitis C (CHC) intercellular and vascular cell adhesion molecules-1 (ICAM-1 and VCAM-1) are expressed de novo on hepatocytes infected by hepatitis C virus and on endothelial cells from sinusoidal vessels, respectively. The soluble forms of these (sICAM-1 and sVCAM-1) reflect their level of expression in tissue. Serum levels of sICAM-1 and VCAM-1 were measured using ELISA assays in 20 patients with CHC, at baseline and after 6 months of treatment with interferon-a + ribavirin. Significantly higher mean values of both adhesions, comparing to healthy controls, were observed. In all patients the lowest value of sICAM-1 was above highest level of controls. At the beginning of the study the responders and nonresponders to treatment (at 1 year) did not differ from each others concerning sICAM-1/sVCAM-1 concentrations. A significant reduction of sICAM-1 levels was apparent after 6 months of therapy, especially in the group of responders. Just a normalization of sICAM-1 values in all but one of responders, comparing to only 5 of 9 nonresponders has been achieved. By contrast, the mean level of sVCAM-1 did not change significantly with therapy. In conclusion, the normalization of serum sICAM-1 at 6 months of treatment may be useful prognostic parameter of response to the end of the administration period (1 year). PMID- 12532198 TI - Results of interferon treatment in children with chronic hepatitis B. AB - Many observations report a variable therapeutical response to interferon in children with chronic hepatitis B. In order to evaluate the efficiency of alpha interferon treatment in the downregulation of viral replication and in the eradication of infection in these patients, we assessed HBeAg/HBeAb and HBsAg/HBsAb seroconversion (as well as with clinical outcome and the changes in the plasma level of aminotransferases) in 61 treated patients. The diagnosis was established by means of the usual clinical, biochemical and histopathological criteria. There was no possibility to viral DNA test and no control group was included. Patients were selected for interferon treatment who displayed at least a two fold rise in the plasma level of aminotransferases as compared to normal values, as well as necroinflammatory activity (score > or = 6) and positive HBeAg as a marker of viral replication. Treatment was carried out with alpha-2a interferon or alpha-2b interferon in a dose of 3 million U/m2/dose in 3 weekly doses for a period of 4-6 months. The monitoring interval was 6.6+/-3 years. HBeAg/HBeAb seroconversion was registered in 77.2% of the patients and mainly occurred during the first year of follow-up (50.9 %). HBsAg/HBsAb seroconversion was revealed in 1.75% of the cases. The therapeutical response was complete, incomplete, transient and absent in 1.75%, 64.9%, 10.5% and 22.8% of the patients, respectively. The results show that the eradication of HBV infection is insignificant, but the downregulation of viral replication and, subsequently the halt of further progression of hepatic lesions is obtained in a high percentage of cases, highlighting the efficiency of this treatment in children with chronic hepatitis B PMID- 12532199 TI - The attitude of Romanian medical trainees towards the eradication of H. pylori. AB - AIM: To evaluate the knowledge of Romanian medical trainees regarding H. pylori infection and its management. METHODS: A specialized questionnaire used in a US survey with similar purpose was distributed to 57 medical interns and residents. The questionnaire included 17 item concerning the testing and treatment of H. pylori infection as well as information sources. RESULTS: Romanian medical interns and residents are generally well informed about diagnostic tests and therapeutic recommendations of national and international guidelines. However the decisions are not always appropriate, in respect to the on site practical possibilities. CONCLUSIONS: Medical faculties should be satisfied with the knowledge of Romanian medical trainees regarding H. pylori infection. However, more emphasis should be given during postgraduate training to the development of practical skills in different specific cases PMID- 12532200 TI - Helicobacter pylori gastritis in children: endoscopical and histological aspects. AB - Helicobacter pylori (Hp) infection is involved in both digestive and extradigestive diseases. Endoscopically, antral location of gastritis, as well as antral nodularity, a consequence of the development of lymphoid follicles in the mucosa of this area, are considered by some authors specific for gastritis in children. Histologically, lymphoid follicles, noticed usually in children, are considered pathognomonic. AIMS: To establish prevalence of gastritis in children and the correlation between of endoscopical and histological findings. PATIENTS AND METHODS: The study group included 267 children in whom upper digestive endoscopy was performed for digestive symptoms. The diagnosis of Hp infection was based on the concordance between the positive urease test and the histological examination (documenting the presence of Hp in the gastric mucosa). RESULTS: Endoscopical aspect of antral nodular gastritis was mainly found in children infected with Hp. Follicular gastritis, considered the histological marker of nodular gastritis, was also revealed in congestive gastritis. Patients infected with Hp most often displayed chronic gastritis and noninfected patients acute gastritis. Hp was not found in gastric mucosa without histological changes. CONCLUSIONS: Nodular pattern of antral mucosa might be considered specific for Hp infection in childhood. In patients infected with Hp chronic gastritis was mainly found while in noninfected children acute gastritis was usually revealed. Hp was not found in gastric mucosa without histological changes. PMID- 12532201 TI - Cystic tumors of the pancreas. Considerations upon 34 operated cases. AB - AIM: To point out the morphologic, clinic and therapeutic aspects of pancreatic cystic tumors. MATERIAL AND METHOD: 34 pancreatic cystic tumors (21 males and 13 females, aged between 21 and 68 years), admitted in the last 15 years were analyzed. They were true cysts in 3 cases (9.9%) and pseudocysts in 31 cases (91.1%), located on the head of the pancreas in 8 cases, on the body in 19, on the tail in 6 and on the body and tail in 1 case. We noticed in the past medical history of the patients with pseudocysts a recent acute pancreatitis attack (26 cases), chronic pancreatitis (4 cases) or a recent abdominal trauma (1 case). The delay between the acute pancreatitis attack and the onset of the pseudocyst varied between 18 days and 2 months. The diagnosis was established by clinical picture (Shefer-Silvis triad), laboratory findings and imaging tests (barium meals, ultrasound test and/or CT test). Thirty cases (27 pseudocysts and 3 true cystic tumors) were operated on: the main surgical procedures were cystogastrostomy (12 cases), cystojejunostomy (6 cases) or cystoduodenostomy (3 cases); we also performed distal pancreatectomy (3 cases), laparostomy or external drainage in 5 cases. RESULTS: We registered 1 death (mortality rate of 3.3%), 2 pancreatic fistulae, 1 pancreatic abscess and 2 recurrences. CONCLUSIONS: 1. The pseudocyst, as an evolutionary complication of acute or chronic pancreatitis, is the most frequent cystic tumor of the pancreas, true pancreatic cysts being extremely rare. 2. The diagnosis is established by clinical pictures, laboratory findings and imaging tests. 3. The treatment is surgical, cystogastrostomy or cystojejunostomy being the main surgical procedures. PMID- 12532202 TI - Salycylate--induced pancreatic injury in the cat: a preliminary study. AB - The effect of intravenous aspirin on the exocrine pancreatic secretion was investigated in a feline isolated pancreaticoduodenal preparation. The study group received a 500 mg/kg bolus dose of aspirin intravenously. Duodenal washouts were collected for six hours. The serum and perfusate aspirin content increased significantly after aspirin administration (p = 0.01). However, the pH, bicarbonate, sodium, potassium and calcium content in the duodenal outflow did not show significant changes between 0 and 6 hours. A significant difference in the perfusate calcium content was present between ASA treated and control cats starting at the end of the first hour of the experiment (p=0.005). Histopathological examination of the pancreas revealed marked erythrocyte extravasation in the ASA treated animals. It is suggested that the ASA- related increase in the calcium secretion of the pancreas should be regarded as an indication of aspirin induced pancreatic damage. PMID- 12532203 TI - Neurogastroenterology--from the basics to the clinics. PMID- 12532204 TI - Autoimmune cholangitis in patients with primary Sjogren's syndrome. A case report. AB - Sjogren's Syndrome is a very frequent autoimmune disease, characterised by exocrine gland involvement. Immunologic disorders are also responsive for extraglandular manifestations of the disease, mostly for the digestive involvement. We report a case of primary Sjogren's syndrome with multiple extraglandular manifestations: vasculitis, cryoglobulinaemia, hepatic involvement and presumably neurologic involvement. The particularities of the case are the typical pattern of autoimmune cholangitis, hypogammaglobulinaemia, the absence of antinuclear antibodies and the association with vasculitis. To the best of our knowledge, this is the first case report of autoimmune cholangitis in Sjogren's syndrome PMID- 12532205 TI - Whipple's disease. Case report. AB - The paper reports the clinical case of a 58 -year-old male patient admitted for diarrhea (6-7 stools/day, diffuse abdominal pain, borborygma, weight loss (20 kgs in two years), asthenia and fatigue. Physical examination evidenced a poor nutritional state (body mass index 19 kg/m2). The abdomen was slightly distended. Biological tests evidenced moderate/severe anemia, hypoproteinemia and hypoalbuminemia. Endoscopic examination evidenced oedematous duodenal mucosa with white-yellowish deposits. Histology (HE stain) revealed the presence of foamy cells and the PAS-staining of the duodenal mucosa evidenced PAS-positive macrophages and numerous intracellular bacilli. Penicillin therapy 2 x 1 million U/day for 14 days, followed by tetracycline 4 x 250 mg/day improved the clinical picture, the patient had only one stool per day and gained weight. After 7 months of treatment the general condition was good and the patient had gained 17 kgs, the duodenal mucosa was normal. HE staining did not evidence foamy cells and no PAS-positive macrophages could be found. PMID- 12532206 TI - Heterotopic bone formation in gastric carcinoma. Case report and discussion of the literature. AB - Osseous metaplasia is a rare finding in the gastrointestinal tract. We report the case of a 75-year-old woman with a gastric adenocarcinoma showing heterotopic ossification with bone marrow formation. The pathogenetic theories are briefly discussed. PMID- 12532207 TI - Ultrasound-guided percutaneous pancreatic biopsy. Indications, performance and complications. AB - The diagnosis of pancreatic masses is often very difficult despite the new developments in imaging methods. As a correct diagnosis has a great impact on the therapy, in certain circumstances it is mandatory to obtain a morphological diagnosis. Although the indications for percutaneous pancreatic biopsy are not standardized, in many circumstances- inoperable pancreatic tumor, suspicion of lymphoma, metastasis or neuroendocrine tumors, differentiation between cystic tumors - a biopsy is clearly indicated. Pancreatic biopsy is usually performed by fine-needles either for cytology or microhistology, the use of larger needles for core biopsy adding a slightly improvement in sensitivity especially in masses from chronic pancreatitis or neuroendocrine tumors. Complications occur seldom, acute pancreatitis and tumoral seeding being the most important. Percutaneous pancreatic biopsy is a safe, flexible and reliable method for diagnosing pancreatic masses PMID- 12532209 TI - [Guidelines for the diagnosis and therapy of primary sclerosing cholangitis. Guidelines of the American College of Gastroenterology 2002]. PMID- 12532210 TI - The Falk symposia. PMID- 12532211 TI - Serotype, mating type and ploidy of Cryptococcus neoformans strains isolated from patients in Brazil. AB - Serotype, mating type and ploidy of 84 strains of Cryptococcus neoformans isolated from 61 AIDS and 23 non-AIDS patients admitted in a tertiary teaching hospital in Sao Paulo, Brazil were examined. Among 61 strains isolated from AIDS patients, 60 strains were var. grubii (serotype A). Only one strain was var. gattii (serotype B). No var. neoformans (serotype D) was found. Among 23 strains isolated from non-AIDS patients, 15 were var. grubii (serotype A) and the remaining 8 were var. gattii, all of which were serotype B. Seventy-three of the 75 serotype A strains were the heterothallic alpha type (MATalpha) and the remaining 2 were untypable (asexual). Most of the MATalpha strains (69/73) were haploid and the remaining 4 strains were diploid. Similarly, both of the 2 asexual strains among the 75 serotype A strains were haploid. There were no alpha mating type (MATalpha) strains among the 84 isolates. All of the 8 var. gattii strains were serotype B and haploid. Among a total of 84 strains tested, neither serotype AD nor serotype D were found. Neither triploid nor tetraploid were found. These results suggest that the serological, sexual and ploidy characteristics in C. neoformans strains isolated from AIDS patients in Sao Paulo were rather simple, whereas strains isolated from non-AIDS patients presented serotype A and B with predominance of serotype A. PMID- 12532212 TI - Human toxocariasis: a seroepidemiological survey in the municipality of Campinas (SP), Brazil. AB - The occurrence of human Toxocara infection was evaluated in three neighborhoods of the periphery of the Campinas municipality (Jardim Santa Monica, Jardim Sao Marcos and Jardim Campineiro) in 1999. Forty residences and 138 residents were randomly selected by drawing lots and were submitted to a seroepidemiological survey, which included blood collection for the immunoenzymatic detection (ELISA) of anti-Toxocara antibodies and a blood count, and the application of a semi structured questionnaire for the evaluation of epidemiological data. Significant levels of anti-Toxocara antibodies were detected in 23.9% of the 1999 samples. No significant difference in the frequency of infection according to age was observed. Environmental contamination with Toxocara eggs was observed in 12.3 and 14.0% of 57 soil samples collected in the same region in December 1998 and July 1999, respectively. Univariate analysis and multiple logistic regression of the data obtained from the questionnaires and of the results of the serological tests, suggest a significant influence of socioeconomic variables on the frequency of human infection with Toxocara under the conditions prevalent in the study area. PMID- 12532213 TI - Prevalence of human herpesvirus 8 antibodies in the population of Belem, Para, Brazil. AB - Serum samples from 497 children and adults inhabiting two neighbourhoods (Guama and Terra Firme) in Belem, Para, North Brazil were screened for the presence of human herpesvirus 8 (HHV-8) antibody using an enzyme-linked immunosorbent assay. An overall 16.3% prevalence was found for these urban communities. Taken both genders together, prevalence rates of HHV-8 antibody increase gradually, across age-groups, ranging from 12.0% to 33.3%. When seroprevalence is analysed by gender, similar rates are found for female (18.4%) and male (14.0%) individuals. In the former gender group, seroprevalence rates increased from 10.3%, in children < or = 10 years of age, to 30.0% in adults 41-50 years of age. Conversely, among male subjects, the prevalence of HHV-8 antibodies decreased from 13.3% in children/young adults aged < or = 10 to 20 years of age to 6.1% in adults aged 21-30 years. From the 31-40 year-old group male onwards, seropositivity rates increased gradually, ranging from 8.3% to 66.7%. A significant difference in seropositivity rates was noted when comparing 21-30 age groups for female and male subjects: 23.3% and 6.1%, respectively (P = 0.03). Geometric mean optical densities were found to increase slightly from the lower to the higher age-groups. Our data suggest that transmission of HHV-8 occurs frequently in the general urban population of Belem, and that prevalence of antibody seems to increase with age. PMID- 12532214 TI - Conventional and molecular typing of Salmonella typhi strains from Brazil. AB - Phenotypic and genotypic characteristics of Salmonella Typhi were studied in 30 strains, isolated in different years, from some areas in Brazil. Conventional typing methods were performed by biochemical tests, Vi phage-typing scheme, and antimicrobial susceptibility test. Molecular typing methods were performed by analysis of plasmid DNA and by random amplified polymorphic DNA (RAPD-PCR). For the latter, an optimization step was performed to ensure the reproducibility of the process in genetic characterization of S. Typhi. The predominance of 76.7% of biotype I (xylose +, arabinose -) was noticed in all studied areas. Three phage types were recognized, with prominence for the phage types A (73.3%) and I+IV (23.3%). All the strains were susceptible to the drugs used. However, 36.7% of the strains contained plasmids, with predominance of the 105 Kb plasmid. RAPD was capable of grouping the strains in 8 genotypic patterns using primer 784, in 6, using primer 787 and in 7, using primer 797. Conventional phenotypic typing methods, as well as the DNA plasmid analysis, presented nonsignificant discriminatory power; however, RAPD-PCR analysis showed discriminatory power, reproducibility, easy interpretation and performance, being considered as a promising alternative typing method for S. Typhi. PMID- 12532215 TI - Botulism: a laboratory investigation on biological and food samples from cases and outbreaks in Brazil (1982-2001). AB - Laboratory investigation of botulism from 1982 to 2001 confirmed the occurrence of eight positive outbreaks/cases of botulism in Brazil. From those, type A botulism was observed in seven of them. Biological material of one case (serum and feces) was positive in the first step of the bioassay, but the amount of sample was not sufficient for typification. One of the outbreaks that occurred in 2001 was negative for botulinum toxin in samples of serum, gastric washing and feces, collected eight days before the onset of the symptoms in the affected person who was clinically diagnosed as presenting the disease. Other two cases presenting compatible clinical diagnoses presented negative results. However, in those cases, the collection of samples was (1) after antiserum administration or (2) later than eight days of the onset of symptoms. Investigation was performed by mouse bioassay, as described in the Compendium of Methods for the Microbiological Examination of Foods (compiled by American Public Health Association--APHA)11, using specific antiserum from Centers for Disease Control (CDC), USA. PMID- 12532216 TI - A prospective study of Cat-Scratch Disease in Lima-Peru. AB - Cat-Scratch Disease (CSD) is a benign lymphadenitis that may progress to severe or recurrent forms, and it is occasionally associated with morbidity. Between January of 1998 and March of 1999, forty-three suspected CSD patients were assessed in the Hospital Cayetano Heredia and the Instituto de Salud del Nino, in Lima, Peru. Twelve patients had a confirmed diagnosis, 8 of whom were women, and the mean age was 10 years old. The majority (53%) of the cases were encountered in the summer. All patients reported having had contact with cats. Fever, malaise, lymphadenopathy and skin lesions were the most frequent clinical features. Twelve patients had indirect immunofluorescence antibody test titers of between 1/50 and 1/800 for Bartonella henselae and Bartonella clarridgeiae. Two lymph node biopsies were histologically compatible with CSD. No positive blood cultures could be obtained. This is the first Peruvian prospective study able to identify B. henselae and B. clarridgeiae in pediatric patients. PMID- 12532217 TI - Seroprevalence of hepatitis B virus infection in individuals with clinical evidence of hepatitis in Goiania, Goias. Detection of viral DNA and determination of subtypes. AB - The presence of serological markers for hepatitis B virus (HBsAg, anti-HBc IgM and Anti-HBc total) was investigated in the serum of 1,396 individuals who had clinical suspect of hepatitis. It was observed that 50.7% of the individuals were positive and, from the total of the studied individuals, 14.5% were positive for HBsAg. From these, 8.5% were also positive for anti-HBc IgM. The analysis in relation to gender showed a higher seroprevalence index among male individuals (p < 0.0001). It was observed the occurrence of subtypes adw2 (62.7%), ayw3 (23.5%), ayw2 (9.8%) and adw4 (3.9%). The viral DNA was detected in 61 (33.9%) HBsAg positive samples and in one sample positive only for anti-HBc total. These results indicate an important incidence of the HBV infection in this population, and reinforce previous studies regarding this virus in the central west region of Brazil. PMID- 12532218 TI - Clinical-epidemiological study of leptospirosis in humans and reservoirs in Yucatan, Mexico. AB - A leptospirosis clinical-epidemiological study was made in humans and reservoirs in the state of Yucatan, Mexico. Interviews and serological analyses were made on 400 persons from an open population, 439 probable cases of leptospirosis and 1060 animal reservoirs (cows, pigs, dogs, rats and opossums). IgM Leptospira Dipstick and Microscopic Agglutination Test (MAT) was used to detect human antibodies to leptospiras and serovar respectively. Leptospirosis incidence in humans was 2.2/100,000 inhab. in 1998, 0.7/100,000 in 1999 and 0.9/100,000 in 2000. Overall seroprevalence was 14.2%, relatively unchanged from seroprevalences observed 20 years ago. Highest seropositivity was found in people over 56 years of age, predominating males over females. Predominant serovars in the open population were tarassovi, hardjo, pomona and panama. Leptospirosis cases were most frequent in rural areas, and the anicteric course predominated over the icteric. The panama, icterohaemorrhagiae and pomona serovars predominated in both anicteric and icteric courses. Dogs, pigs and rodents had the highest seropositivity among the reservoirs. Contact with rodents and natural water sources were significant factors (p < or = 0.05). Human cases (74%) occurred during the rainy season. It is concluded that leptospirosis is still a serious illness with important clinical and epidemiological implications in the state of Yucatan, Mexico. PMID- 12532219 TI - Perinephric and renal abscesses in children: a study of three cases. AB - Perinephric and renal abscesses are rare in childhood, but they must be remembered in the differential diagnosis of fever and abdominal or flank pain. This paper reports three cases whose solution was found after clinical and surgical treatment. The importance of ultrasound and computerized tomography (CT) in prompt diagnosis is highlighted in this work. The use of these methods along the improved antimicrobial therapy may result in decrease in mortality due to this pathology. PMID- 12532220 TI - Brazilian journal of medical and biological research 1981-2002. PMID- 12532221 TI - Pathogenesis of Salmonella-induced enteritis. AB - Infections with Salmonella serotypes are a major cause of food-borne diseases worldwide. Animal models other than the mouse have been employed for the study of nontyphoidal Salmonella infections because the murine model is not suitable for the study of Salmonella-induced diarrhea. The microbe has developed mechanisms to exploit the host cell machinery to its own purpose. Bacterial proteins delivered directly into the host cell cytosol cause cytoskeletal changes and interfere with host cell signaling pathways, which ultimately enhance disease manifestation. Recently, marked advances have been made in our understanding of the molecular interactions between Salmonella serotypes and their hosts. Here, we discuss the molecular basis of the pathogenesis of Salmonella-induced enteritis. PMID- 12532222 TI - The conservative physiology of the immune system. AB - Current immunological opinion disdains the necessity to define global interconnections between lymphocytes and regards natural autoantibodies and autoreactive T cells as intrinsically pathogenic. Immunological theories address the recognition of foreignness by independent clones of lymphocytes, not the relations among lymphocytes or between lymphocytes and the organism. However, although extremely variable in cellular/molecular composition, the immune system preserves as invariant a set of essential relations among its components and constantly enacts contacts with the organism of which it is a component. These invariant relations are reflected, for example, in the life-long stability of profiles of reactivity of immunoglobulins formed by normal organisms (natural antibodies). Oral contacts with dietary proteins and the intestinal microbiota also result in steady states that lack the progressive quality of secondary-type reactivity. Autoreactivity (natural autoantibody and autoreactive T cell formation) is also stable and lacks the progressive quality of clonal expansion. Specific immune responses, currently regarded as the fundament of the operation of the immune system, may actually result from transient interruptions in this stable connectivity among lymphocytes. More permanent deficits in interconnectivity result in oligoclonal expansions of T lymphocytes, as seen in Omenn's syndrome and in the experimental transplantation of a suboptimal diversity of syngeneic T cells to immunodeficient hosts, which also have pathogenic consequences. Contrary to theories that forbid autoreactivity as potentially pathogenic, the physiology of the immune system is conservative and autoreactive. Pathology derives from failures of these conservative mechanisms. PMID- 12532223 TI - p53 as a prognostic factor in adrenocortical tumors of adults and children. AB - Mutations of the tumor suppressor gene p53 have been considered to be important determinants in several kinds of human cancer. Accumulation of p53 protein has been reported to correlate with more aggressive clinical behavior in some neoplasms. The role of p53 expression in adrenal cortical tumors (ACT) has not been elucidated but some studies have suggested its correlation with malignant behavior. Our objective was to determine if there is a correlation between the expression of immunoreactive p53 and the biological behavior of ACT. Fifty-seven ACT (21 from children and 36 from adults) were evaluated for p53 expression by immunohistochemistry in formalin-fixed paraffin-embedded tissue and analyzed in terms of outcome. The p53 parameter was utilized semiquantitatively. Tumors were classified as p53 negative when no positivity was observed, or when only few cells showed weak positivity (0/1+) and scored as p53 positive when there was a diffuse and strong nuclear positivity (2+/3+). In children, p53 positivity was associated with clinically malignant ACT and p53 negativity was associated with clinically benign ACT (P = 0.026). In adults' ACT, p53 positivity had an effect on disease-free survival (P<0.001) and also correlated with Weiss score, with a cutoff = 4 (P = 0.04). p53 expression was related to the clinical behavior of ACT in both children and adults and these findings seem to support a role for p53 in ACT progression. PMID- 12532224 TI - Sec61alpha synthesis is enhanced during translocation of nascent chains of collagen type IV in F9 teratocarcinoma cells after retinoic acid treatment. AB - Nascent procollagen peptides and other secretory proteins are transported across the endoplasmic reticulum (ER) membrane through a protein-conducting channel called translocon. Sec61alpha, a multispanning membrane translocon protein, has been implicated as being essential for translocation of polypeptide chains into the cisterns of the ER. Sec61alpha forms a protein complex with collagen and Hsp47, an ER-resident heat shock protein that binds specifically to collagen. However, it is not known whether Sec61alpha is ubiquitously produced in collagen producing F9 teratocarcinoma cells or under heat shock treatment. Furthermore, the production and utilization of Sec61alpha may depend on the stage of cell differentiation. Cultured F9 teratocarcinoma cells are capable of differentiation in response to low concentrations of retinoic acid. This differentiation results in loss of tumorigenicity. Mouse F9 cells were grown in culture medium at 37 degrees C and 43 degrees C (heat shock treatment) treated or not with retinoic acid, and labeled in certain instances with 35S-methionine. Membrane-bound polysomes of procollagen IV were then isolated. Immunoprecipitation and Western blot analysis were performed using polyclonal antibodies against collagen IV, Hsp47 and Sec61alpha. Under retinoic acid-untreated conditions, F9 cells produced undetectable amounts of Sec61alpha. Sec61alpha, Hsp47 and type IV collagen levels were increased after retinoic acid treatment. Heat shock treatment did not alter Sec61alpha levels, suggesting that Sec61alpha production is probably not affected by heat shock. These data indicate that the enhanced production of Sec61alpha in retinoic acid-induced F9 teratocarcinoma cells parallels the increased synthesis of Hsp47 and collagen type IV. PMID- 12532225 TI - Slaughterhouse blood as a perfusate for studying myocardial function under ischemic conditions. AB - Metabolic studies using the in vitro non-recirculating blood-perfused isolated heart model require large volumes of blood. The present study was designed to determine whether heterologous pig blood collected from a slaughterhouse can be used as perfusate for isolated pig hearts perfused under aerobic and constant reduced flow conditions. Eight isolated working pig hearts perfused for 90 min at a constant flow of 1.5 ml g(-1) min(-1) with non-recirculated blood diluted with Krebs-Henseleit bicarbonate buffer at a hematocrit of 23% were compared to eight hearts subjected to the same protocol but perfused only with Krebs-Henseleit bicarbonate buffer solution. Hearts were paced at 100 bpm and subjected to aerobic perfusion at 38 degrees C. Hearts were weighed before perfusion and at the end of the experiment and the results are reported as percent weight gain (mean +/- SD). Comparisons between groups were performed by the Student t-test (P<0.05). After 90 min of perfusion with modified Krebs-Henseleit, perfused hearts presented a larger weight gain than blood-perfused hearts (39.34 +/- 9.27 vs 23.13 +/- 5.42%, P = 0.003). Left ventricular end-diastolic pressure was higher in the modified Krebs-Henseleit-perfused group than in the blood group (2.8 +/- 0.4 vs 2.3 +/- 0.3 mmHg, respectively, P = 0.01). We conclude that heterologous blood perfusion, by preserving a more physiological myocardial water content, is a better perfusion fluid than modified Krebs-Henseleit solution for quantitative studies of myocardial metabolism and heart function under ischemic conditions. PMID- 12532226 TI - Responsiveness of glycogen breakdown to cyclic AMP in perfused liver from rats with insulin-induced hypoglycemia. AB - The responsiveness of glycogen breakdown to cAMP was investigated in isolated perfused liver from male Wistar fed rats (200-220 g) with insulin-induced hypoglycemia. The activation of glycogenolysis by 3 microM cAMP was decreased (P<0.05) in livers from rats with hypoglycemia induced by the administration of insulin or during the direct infusion of insulin into the isolated liver. The direct effect of insulin on glycogen catabolism promoted by 3 microM cAMP occurred as early as 3 min after starting insulin infusion. In contrast, the cAMP agonists resistant to phosphodiesterases, 8Br-cAMP and 6MB-cAMP, used at the same concentration as cAMP, i.e., 3 microM, did not modify the effect of insulin. The data suggest that the decreased hepatic responsiveness of glycogen breakdown during insulin-induced hypoglycemia is a direct effect of insulin decreasing the intracellular levels of cAMP. PMID- 12532227 TI - Changes in endogenous tissue glutathione level in relation to murine ascites tumor growth and the anticancer activity of cisplatin. AB - Changes in glutathione levels were determined in tissues of 11- to 12-week-old Swiss albino mice at different stages of Dalton's lymphoma tumor growth and following cisplatin (8 mg/kg body weight, ip) treatment for 24-96 h, keeping 4-5 animals in each experimental group. Glutathione levels increased in spleen of tumor-bearing compared to normal mice (9.95 +/- 0.14 vs 7.86 +/- 1.64 micromol/g wet weight, P69.5 kb) and human USP15 gene (145 kb) sequences in genome databases reveals that both are composed of 22 exons with identical splice sites, and both have an exon/intron structure identical to the mouse Usp4 gene, including the alternately spliced exon. Phylogenetic studies suggest that a sequence currently identified as a chicken Usp4 ortholog is in fact a USP15 ortholog, while bona-fide chicken, cow, and rat Usp4 orthologs can be identified in EST databases. PMID- 12532267 TI - A study of short utrophin isoforms in mice deficient for full-length utrophin. AB - Utrophin can functionally replace dystrophin in dystrophin-deficient muscle and may have a role in a therapeutic strategy for Duchenne muscular dystrophy. This has resulted in many investigations of the full-length muscle form of utrophin; however, the short utrophins and non-muscle forms have been relatively neglected, partly because they are difficult to analyze in the presence of the full-length form. Our study circumvents this problem by using mice deficient for the full length form (UKOex6 mice) to study the translation and distribution of short utrophins. Four tissues were examined-kidney, testis, fetal hands/feet, and brain and three novel short isoforms were identified, including Up120, which appears to be specific to kidney glomeruli, and Up 109, expressed in the fetal dermis. A third form, Up103, was found in testis but at extremely low levels. A cDNA for Up109 has been isolated and shown to have a unique NH2-terminal sequence. In addition, the first exons of Up109 and another short form, G-utrophin, have both been located within intron 55, 56 kb apart. Our immunological studies show that G utrophin protein accumulates only in neural tissue, in line with its similarly restricted RNA distribution. Our study of testis expression shows, for the first time, that full-length utrophin is expressed at high levels in Leydig cells, raising the possibility that this protein is involved in testosterone secretion. We note that translation of the short utrophins, especially Up140 and Up71, is relatively inefficient and discuss the significance of this observation. PMID- 12532268 TI - Phylogenetics of rat inbred strains. PMID- 12532270 TI - Rearranged gene order between pig and human in a QTL region on SSC 7. AB - On porcine Chromosome 7, the region surrounding the MHC region contains QTL influencing many traits including growth, back fat thickness, and carcass composition. Towards the identification of the responsible gene(s), this article describes an increase of density of the radiated hybrid map of SSC 7 in the q11 q14 region and the comparative analysis of gene order on the porcine RH map and human genome assembly. Adding 24 new genes in this region, we were able to build a framework map that fills in gaps on the previous maps. The new software Carthagene was used to build a robust framework in this region. Comparative analysis of human and porcine maps revealed a global conservation of gene order and of distances between genes. A rearranged fragment of around 3.7 Mb was, however, found in the pig approximately 20 Mb upstream from the expected location on the basis of the human map. This rearrangement, found by RH mapping on the IMpRH 7.000 rads panel, has been confirmed by two-color FISH and by mapping on the high resolution IMNpRH2 12.000 rads panel. The rearranged fragment contains two microsatellites found at the most likely QTL location in the INRA QTL experiment. It also contains the BMP5 gene, which, together with CLPS, could be considered as a possible candidate. PMID- 12532271 TI - Sex and strain-related differences in the peripheral blood cell values of inbred mouse strains. PMID- 12532272 TI - Function and regulation of the mammalian copper-transporting ATPases: insights from biochemical and cell biological approaches. AB - Copper is an essential trace element that plays a very important role in cell physiology. In humans, disruption of normal copper homeostasis leads to severe disorders, such as Menkes disease and Wilson's disease. Recent genetic, cell biological, and biochemical studies have begun to dissect the molecular mechanisms involved in transmembrane transport and intracellular distribution of copper in mammalian cells. In this review, we summarize the advances that have been made in understanding of structure, function, and regulation of the key human copper transporters, the Menkes disease and Wilson's disease proteins. PMID- 12532274 TI - Trafficking of Na,K-ATPase fused to enhanced green fluorescent protein is mediated by protein kinase A or C. AB - Fusion of enhanced green fluorescent protein (EGFP) to the C-terminal of rat Na,K ATPase a1-subunit is introduced as a novel procedure for visualizing trafficking of Na,K-pumps in living COS-1 renal cells in response to PKA or PKC stimulation. Stable, functional expression of the fluorescent chimera (Na,K-EGFP) was achieved in COS-1 cells using combined puromycin and ouabain selection procedures. Na,K pump activities were unchanged after fusion with EGFP, both in basal and regulated states. In confocal laser scanning and fluorescence microscopes, the Na,K-EGFP chimera was distributed mainly along the plasma membrane of COS cells. In unstimulated COS cells, Na,K-EGFP was also present in lysosomes and in vesicles en route from the endoplasmic reticulum to the plasma membrane, but it was almost absent from recycling endosomes labelled with fluorescent transferrin. After activation of protein kinase A or C, the density of co-localizing Na,K-EGFP and transferrin vesicles was increased 3-4-fold, while the ouabain-sensitive 86Rb uptake was reduced by 22%. Simultaneous activation of PKA and PKC had additive effects with a 6-fold increase of co-localization and a 38% reduction of 86Rb uptake. Responses of similar magnitude were seen after inhibition of protein phosphatase by okadaic acid. Reduction of the amount of Na,K-ATPase in surface plasma membranes through internalization in recycling endosomes may thus in part explain a decrease in Na,K-pump activity following protein kinase activation or protein phosphatase inhibition. PMID- 12532273 TI - P-type ATPases in Caenorhabditis and Drosophila: implications for evolution of the P-type ATPase subunit families with special reference to the Na,K-ATPase and H,K-ATPase subgroup. AB - Here we show a complete list of the P-type ATPase genes in Caenorhabditis elegans and Drosophila melanogaster. A detailed comparison of the deduced amino-acid sequences in combination with phylogenetic and chromosomal analyses has revealed the following: (1) The diversity of this gene family has been achieved by two major evolutionary steps; the establishment of the major P-type ATPase subgroups with distinct substrate (ion) specificities in a common ancestor of vertebrate and invertebrate, followed by the evolution of multiple isoforms occurring independently in vertebrate and invertebrate phyla. (2) Pairs of genes that have intimate phylogenetic relationship are frequently found in proximity on the same chromosome. (3) Some of the Na,K- and H,K-ATPase isoforms in D. melanogaster and C. elegans lack motifs shown to be important for alpha/beta-subunit assembly, suggesting that such alpha- and beta-subunits might exist by themselves (lonely subunits). The mutation rates for these subunits are much faster than those for the subunits with recognizable assembly domains. (4) The lonely alpha-subunits also lack the major site for ouabain binding that apparently arose before the separation of vertebrates and invertebrates and thus well before the separation of vertebrate Na,K-ATPases and H,K-ATPases. These findings support the idea that a relaxation of functional constraints would increase the rate of evolution and provide clues for identifying the origins of inhibitor sensitivity, subunit assembly, and separation of Na,K- and H,K-ATPases. PMID- 12532275 TI - Differential effect of omega3 PUFA supplementations on Na,K-ATPase and Mg-ATPase activities: possible role of the membrane omega6/omega3 ratio. AB - Several functional properties of Na,K-ATPase are strongly dependent on membrane fatty acid composition, but the underlying mechanism is still not well defined. We have studied the effects of two types of supplementations enriched in the w3 polyunsaturated fatty acids on the Na,K-ATPase and Mg-ATPase activities in sciatic nerve (SN) and red blood cells (RBC). Eight groups of rats, controls and diabetics, received a standard diet, supplemented or not with 30 or 60 mg/kg/day of docosahexaenoic acid (DHA) or with soybean for eight weeks. Diabetes induced significant decrease of Na,K-ATPase activity in SN (-23%) and RBC (-25%), without affecting Mg-ATPase activity. In RBC, soybean and DHA supplementations caused significant increases in Na,K-ATPase activity (in various range, +13% to +145%) in all groups, and in Mg-ATPase activity in control soybean (+65%), control and diabetic DHA high dose (+39%, +53%) and diabetic DHA low dose (+131%) groups. In SN, the soybean caused a significant decrease in Na,K-ATPase activity (-26%) and still more in the diabetic group (-53%). The DHA diet induced a slight decrease in activity in control groups, whilst during diabetes, at high dose, we noted an aggravation of this decrease (-36%). Mg-ATPase activity was not modified by supplementations except for the low dose of DHA where the activity was slightly decreased in the control group (-16%). The supplementations induced multiple tissue-specific modifications in the membrane fatty acid composition of RBC and of SN homogenates. Several specific correlations have been found between variations in fatty acids amounts and Na,K-ATPase activity in these tissues but only in RBC for Mg-ATPase activity. Indeed, we observed that the variations in Na,K-ATPase activity are positively and significantly correlated with changes in the omega6/omega3 ratio in SN as well as in RBC. These data clearly show, for the first time, that the diet could modulate the Na,K-ATPase activity via the omega6/omega3 ratio in the membranes. A similar correlation was observed with Mg ATPase activity in RBC, suggesting also a dietary regulation of the enzyme; but for the SN, this activity might be regulated by a different omega6/omega3 ratio or by another pathway. PMID- 12532276 TI - Two histidine residues in the juxta-membrane cytoplasmic domain of Na+/H+ exchanger isoform 3 (NHE3) determine the set point. AB - Histidine residues in Na+/H+ exchangers are believed to participate in proton binding and influence the Na+/H+ exchanger activity. In the present study, the function of three highly conserved histidines in the juxtamembrane cytoplasmic domain of NHE3 was studied. His-479, His-485, and His-499 were mutated to Leu, Gln or Asp and expressed in an Na+/H+ exchanger null cell line and functional consequences on Na+/H+ exchange kinetics were characterized. None of the histidines were essential for NHE3 activity, with all mutated NHE3 resulting in functional exchangers. However, the mutation in His-475 and His-499 significantly lowered NHE3 transport activity, whereas the mutation in H485 showed no apparent effect. In addition, the pH profiles of the H479 and H499 mutants were shifted to a more acidic region, and lowered its set point, the intracellular pH value above which the Na+/H+ exchanger becomes inactive, by approximately 0.3-0.6 pH units. The changes in set point by the mutations were further shifted to more acidic values by ATP depletion, indicating that the mechanism by which the mutations on the histidine residues altered the NHE3 set point differs from that caused by ATP depletion. We suggest that His-479 and His-499 are part of the H+ sensor, which is involved in determining the sensitivity to the intracellular H+ concentration and Na+/H+ exchange rate. PMID- 12532269 TI - Generation and sequence characterization of a normalized cDNA library from swine ovarian follicles. AB - Ovulation rate is a major factor determining litter size in swine and is, therefore, a trait of economic importance to the pork industry. The dynamics of follicle development, which in turn are dictated by a balance between follicle recruitment, maturation, selection, and atresia, are a major determining factor of ovulation rate. The role of several genes expressed in the ovaries during these processes has been described, but studies utilizing large-scale genomic approaches have yet to be conducted to examine gene expression in this tissue more globally. We have developed a normalized cDNA library from swine ovarian follicles in various stages of development, ranging from 2.0 to 10.0 mm in diameter, collected from gilts from divergent genetic lines selected for high and low ovulation rates, during the 7 initial days of the follicular phase of the estrous cycle. EST sequences were obtained from 5231 distinct clones derived from this library. In total, 3479 unique sequence clusters were obtained, of which 2661 singletons (76.5%) were observed. BLASTN searches with the primary sequences from the clusters obtained resulted in 1037 sequences not matching (E <1.0(-06) any of the sequences in the nt database (29.8% novelty rate). This resource will facilitate the use of cDNA microarrays in functional genomics studies aiming at unraveling the genetic and physiological mechanisms underlying follicle maturation and ovulation rate in swine. PMID- 12532277 TI - 17beta-estradiol and tamoxifen regulate a maxi-chloride channel from human placenta. AB - Steroid hormones have been implicated in the modulation of several transport processes, including conductive chloride transport in epithelial cells. Micromolar concentrations of these hormones have been determined in blood of pregnant women. The purpose of this work was to explore the effects of 17beta Estradiol, a steroid hormone, on the biophysical properties of the Maxi chloride channel present in apical membranes from human placental syncytiotrophoblast. Apical membrane chloride channels from human term placentas were reconstituted in giant liposomes suitable for electrophysiologic studies by the patch-clamp method. Low micromolar concentrations of 17beta-Estradiol inhibit the Maxi chloride channels in excised patches in a potential-dependent manner. The addition of 1 mM 17beta-Estradiol to the bath solution decreased the total current in the patch from 100% control to 71% at -40 mV holding potential and the current was not affected by 17beta-Estradiol at + 40 mV. However, the presence of the hormone did not affect the single-channel conductance, therefore its effect must be due to modulation of its open probability (Po). Interestingly, 17alpha Estradiol did not change the total current in the patch. Tamoxifen, an antiestrogen, also showed inhibition, but in a voltage-independent manner. Our results suggest that the Maxi Cl- channel from human term placenta may be regulated by direct interaction of both compounds with the channel. From a functional point of view, the control of these channels by steroid hormones may be of great importance in placental physiology and their regulation may help to unravel their possible role in transplacental transport. PMID- 12532278 TI - Removal of the MDCK cell primary cilium abolishes flow sensing. AB - The hypothesis that cell primary cilium is solely responsible for the flow induced Ca2+ response in MDCK cells was tested by removal of the cilia from mature, responsive cells. Incubation of the cells with 4 mM chloral hydrate for 68 hours resulted in the complete loss of the primary cilia and in disorganization of microtubules, as visualized by immunofluorescence. When intracellular Ca2+ concentration was measured with Fluo-4, the elevation that normally accompanies an increase in fluid flow was abolished after 20 hours exposure to chloral hydrate. At this time, the primary cilia still remained attached to the cells but had become twisted and flexible. Twenty-four hours after return of the deciliated cells to normal medium, intracellular microtubule organization appeared normal, but primary cilia had not yet been expressed. The cells failed to increase intracellular Ca2+ in response to fluid flow until after they had been in normal medium for 120 hours, at which time the primary cilia were 3-4 microm long. Chloral hydrate did not impair the Ca2+ mobilization machinery, as the Ca2+ response to mechanical contact and the spread to neighboring cells was unaffected by the drug. We conclude that the primary cilium is the only sensor for the flow-induced Ca2+ response in MDCK cells and estimate that a single mechanically sensitive channel in the cilium could provide the requisite Ca2+ influx. PMID- 12532279 TI - Hepatocyte growth factor/scatter factor stimulates Ca2+-activated membrane K+ current and migration of MDCK II cells. AB - Hepatocyte growth factor/scatter factor (HGF/SF) stimulates migration of various cells and has been linked via Met tyrosine kinase-signaling to transformation and the metastatic phenotype. Migration of transformed MDCK-F cells depends on activation of a charybdotoxin-sensitive, volume-activated membrane K+ current. Thus, we used patch-clamp electrophysiology and transwell migration assays to determine whether HGF/SF stimulation of MDCK II cell migration depends on the activation of membrane K+ currents. HGF/SF activated a membrane K+ current that increased over 24 hr, and which could be modulated by increasing intracellular calcium concentration, [Ca2+]i. Charybdotoxin (ChTX, 50 nM), iberiotoxin (IbTX, 100 nM), stichodactyla toxin (Stk, 100 nM) and clotrimazole (CLT, 1 mM) all inhibited this current. HGF/SF (100 scatter units/ml) significantly increased MDCK II cell migration over 8 hr compared to control cells. Addition of ChTX (50 nM), IbTX (100 nM), Stk (100 nM) or CLT (1 microM) inhibited the HGF/SF stimulated MDCK II cell migration. We conclude that the activation of membrane Ca2+-activated K+current is necessary for HGF/SF stimulation of MDCK II cell. PMID- 12532280 TI - Effect of fluoride pretreatment on the solubility of synthetic carbonated apatite. AB - The purpose of this research was to address the following question: How is the solubility of fluoride-pretreated carbonated apatite (CAP) in aqueous acidic media related to the equilibrium solution fluoride and/or the CAP adsorbed fluoride levels? A CAP sample prepared by a precipitation method at 70 degrees C containing approximately 6% carbonate was fluoride-treated (F adsorption from neutral aqueous solutions) to yield a approximately 1000 ppm F CAP and a approximately 3300 ppm F CAP. Metastable equilibrium solubility distributions were determined in acetate buffers at pH 5.0. Solution fluoride, calcium, phosphate, and pH were determined from the equilibrated solutions. The equilibrium solution fluoride levels were extremely low, e.g., as low as approximately 0.10 ppb to approximately 0.30 ppb at 50% dissolved for the two CAP preparations. The approximately 3300 ppm F CAP yielded a lower solubility than the approximately 1000 ppm F CAP (shift in the mean pKHAP value of 1.5-2 units). This can be attributed to the lower solution F(-) for the sample containing approximately 1000 ppm fluoride compared with the approximately 3300 ppm fluoride containing CAP. These important findings suggest that a fluoride treatment simply may provide an adsorption fluoride depot for subsequent release, providing a solution fluoride effect upon the CAP solubility and not necessarily any intrinsic alteration of the mineral solubility. PMID- 12532281 TI - The ratio of osteocytic incorporation to bone matrix formation in femoral neck cancellous bone: an enhanced osteoblast work rate in the vicinity of hip osteoarthritis. AB - Recently it has been shown that an inactivating mutation in the TGFb-SMAD3 signaling pathway, which increases the conversion of osteoblasts to osteocytes, is accompanied by bone loss combined with increased osteocyte density. We hypothesized that increased matrix TGFb, known to occur in osteoarthritis, might cause the reverse of these effects in man. Because coxarthrosis (cOA) is associated with a reduced risk of femoral neck fracture, whole cross-section femoral neck biopsies were obtained from 11 patients with femoral neck fracture, 14 patients with cOA, and 22 age-and sex-matched controls. Lacunar density (Lc x mm2), osteocyte density (Ot x mm2), and cancellous wall width (Cn x W x Wi), were compared between cases of coxarthrosis, femoral neck fracture (FNF) and controls. In cOA, Lc.mm2 was reduced by 24% (P <0.001) while in FNF it was increased by 20% (P <0.001). Cn x W x Wi was increased in cOA by 22% (P <0.05) and in FNF was reduced by 27% (P <0.001). Lc x mm2 was inversely related to percentage cancellous bone area (adj. r2 = 0.373; P <0.01) and wall widths, r2 = 0.382, P <0.001. The reduction in osteocyte lacunar density coupled with increased wall width is consistent with a model of cOA effects on bone in which increased levels of matrix TGFb might prolong the effective lifespan or work rate of the osteoblast and delay its incorporation into the matrix as an osteocyte. One possible approach to strengthening bone in osteoporosis might be to enhance the effective lifespan of the osteoblast by modulating TGFb-related pathway activity in its local environment. PMID- 12532282 TI - Mechanical control of human osteoblast apoptosis and proliferation in relation to differentiation. AB - Bone cells respond to mechanical stimulation. This is thought to be the mechanism by which bone adapts to mechanical loading. Reported responses of bone cells to mechanical stimuli vary widely and therefore there is no consensus on what mechanisms of mechanotransduction are physiologically relevant. We hypothesize that the differentiation stage of osteoblastic cells used to study responses to strain in vitro determines the outcome of applied loading. A human fetal osteoblast cell line was triggered to differentiate in culture to the advanced state of mineralization by addition of the osteogenic factors dexamethasone and b glycerophosphate. Osteoblast cultures were subjected to increasing levels of cyclic, equibiaxial stretch at different stages of differentiation. We show that differentiation of human osteoblasts affects their responses to stretch in vitro. In 7-day osteoblast cultures, stretch results in decreased cell numbers as cells are triggered into apoptosis, independent of the stretch level (between 0.4 2.5%). In more mature cultures, apoptosis is not affected by the same treatment. Stretching differentiating cultures at day 14 actually increases proliferation. This is the first study reporting on differentiation-dependent mechanical control of osteoblast proliferation and apoptosis and is fundamental in understanding mechanotransduction processes in bone. The tight regulation of these responses by differentiation implies the significance of the differentiation stage of osteoblasts for the translation of mechanical signals and corroborates with the putative role of the osteoblastic lineage as mechanotransducer in bone. PMID- 12532285 TI - Superbeads: immobilization in "sweet" chemistry. AB - Enzymatic oligosaccharide synthesis using recombinant glycosyltransferases is able to overcome the difficulties associated with chemical methods. Nonetheless, sugar nucleotide regeneration cycles are necessary for the glycosylation. The multistep enzyme reaction can be efficiently carried out on superbeads that are prepared by immobilizing multienzyme mixtures on bead support through fused binding domains. PMID- 12532283 TI - Reduced Y-chromosome, but not mitochondrial DNA, diversity in human populations from West New Guinea. AB - To investigate the paternal population history of New Guinea, 183 individuals from 11 regional populations of West New Guinea (WNG) and 131 individuals from Papua New Guinea (PNG) were analyzed at 26 binary markers and seven short-tandem repeat loci from the nonrecombining part of the human Y chromosome and were compared with 14 populations of eastern and southeastern Asia, Polynesia, and Australia. Y-chromosomal diversity was low in WNG compared with PNG and with most other populations from Asia/Oceania; a single haplogroup (M-M4) accounts for 75% of WNG Y chromosomes, and many WNG populations have just one Y haplogroup. Four Y chromosomal lineages (haplogroups M-M4, C-M208, C-M38, and K-M230) account for 94% of WNG Y chromosomes and 78% of all Melanesian Y chromosomes and were identified to have most likely arisen in Melanesia. Haplogroup C-M208, which in WNG is restricted to the Dani and Lani, two linguistically closely related populations from the central and western highlands of WNG, was identified as the major Polynesian Y-chromosome lineage. A network analysis of associated Y chromosomal short-tandem-repeat haplotypes suggests two distinct population expansions involving C-M208--one in New Guinea and one in Polynesia. The observed low levels of Y-chromosome diversity in WNG contrast with high levels of mtDNA diversity reported for the same populations. This most likely reflects extreme patrilocality and/or biased male reproductive success (polygyny). Our data further provide evidence for primarily female-mediated gene flow within the highlands of New Guinea but primarily male-mediated gene flow between highland and lowland/coastal regions. PMID- 12532286 TI - A process in need is a process indeed: scalable enantioselective synthesis of chiral compounds for the pharmaceutical industry. AB - This report deals with enantioselective synthesis of viracept 1 (nelfinavir mesylate, AG 1343), a potent HIV protease inhibitor, and 3-hydroxytetradecanoic acid 3, a component of lipid A comprising lipopolysaccharide embedded in the cell surface of Gram-negative bacteria, from both strategic and practical perspectives. As regards the synthesis of 1, the synthetic approaches to its central intermediate 2 possessing the common structural motif of 1,4 differentially substituted-2-amino-3-hydroxylbutane are mainly discussed with emphasis on the molecular symmetry that has helped streamline the synthetic strategy. In the discussion of the synthetic strategies to access a single enantiomer of 3, the chiral methodologies that have been applied so far are assessed for industrial viability; the synthetic alternatives explored include resolution via diastereomeric salt formation, lipase-catalyzed kinetic resolution, asymmetric synthesis, and chiral pool approaches. PMID- 12532287 TI - Total synthesis of the antitumor acetogenin mosin B: desymmetrization approach to the stereodivergent synthesis of threo/trans/erythro-type acetogenins. AB - A total synthesis of the threo/trans/erythro-type acetogenin mosin B and one of its diastereomers has been achieved. The carbon skeleton is assembled in a convergent fashion from two segments (a THF ring segment and a gamma-lactone segment) through the Nozaki-Hiyama-Kishi reaction. The THF ring segment was stereoselectively constructed by a stereodivergent synthesis starting from a common intermediate (4-cyclohexene-1,2-diol) based on a desymmetrization strategy. The gamma-lactone segment was synthesized by coupling a triflate and a chiral alpha-sulfenyl gamma-lactone. By virtue of these synthetic results, we suggest that the absolute configuration of natural mosin B is 1 a. Antiproliferative effects of 1 a and 1 b were also investigated. PMID- 12532288 TI - The delocalization index as an electronic aromaticity criterion: application to a series of planar polycyclic aromatic hydrocarbons. AB - This work introduces a new local aromaticity measure, defined as the mean of Bader's electron delocalization index (DI) of para-related carbon atoms in six membered rings. This new electronic criterion of aromaticity is based on the fact that aromaticity is related to the cyclic delocalized distribution of pi electrons. We have found that this DI and the harmonic oscillator model of aromaticity (HOMA) index are strongly correlated for a series of six-membered rings in eleven planar polycyclic aromatic hydrocarbons. The correlation between the DI and the nucleus-independent chemical shift (NICS) values is less remarkable, although in general six-membered rings with larger DI values also have more negative NICS indices. We have shown that this index can also be applied, with some modifications, to study of the aromaticity in five-membered rings. PMID- 12532289 TI - Metal-free, noncovalent catalysis of diels-alder reactions by neutral hydrogen bond donors in organic solvents and in water. AB - We examined the catalytic activity of substituted thioureas in a series of Diels Alder reactions and 1,3-dipolar cycloadditions. The kinetic data reveal that the observed accelerations in the relative rates are more dependent on the thiourea substituents than on the reactants or solvent. Although the catalytic effectiveness is the strongest in noncoordinating, nonpolar solvents, such as cyclohexane, it is also present in highly coordinating polar solvents, such as water. In 1,3-dipolar cycloadditions, the thiourea catalysts demonstrate only very moderate selectivity for reactions with inverse electron demand. Our experiments emphasize that both hydrophobic and polar interactions can co-exist, making these catalysts active, even in highly coordinating solvents. This class of catalysts increases the reaction rates and endo-selectivities of Diels-Alder reactions, in a similar manner to weak Lewis acids, without concomitant product inhibition. PMID- 12532290 TI - Structural studies of N,N'-di(ortho-fluorophenyl)formamidine group 1 metallation. AB - The treatment of N,N'-di(ortho-fluorophenyl)formamidine (HFPhF) in tetrahydrofuran with equimolar amounts of n-butyllithium, sodium bis(trimethylsilyl)amide or potassium bis(trimethylsilyl)amide affords the colourless crystalline formamidinate complexes [Li(FPhF)(thf)] (1), [Na(FPhF)(thf)] (2) and [K(FPhF)] (5). Low-temperature preparation of 2 in diethyl ether yields the Et(2)O adduct [Na(FPhF)(Et(2)O)] (3). At ambient temperature the sodium fluoride inclusion complex [Na(3)(FPhF)(3)(Et(2)O)(NaF)] (4) is also formed. Spectroscopic ((1)H, (13)C and (19)F((1)H) NMR) data for 1-5, microanalytical analyses for compounds 1, 2 and 5 and X-ray structure determinations for 1, 3-5 confirm the formulae of these species. In the solid state, 1 and 3 possess a dimeric nature in which the formamidinate ligands coordinate through mu(2):eta(2):eta(1) (1) and mu(2):eta(2):eta(2) (3) binding modes. These are enabled by partial ortho-fluoro donation. Compound 4, which is also dimeric, contains two trisodium tris(formamidinate) units that comprise mu(2):eta(2):eta(2)-FPhF ligands, a bridging diethyl ether moiety and an unprecedented mu(3):eta(2):eta(2):eta(2)-formamidinate donor. Together, these trinuclear units encapsulate two sodium fluoride units by eta(2)-N,N formamidinate chelation of the sodium cations (thereby creating further mu(3):eta(2):eta(2):eta(2)-bound formamidinates) and fluoride-sodium interactions. Compound 5 extends the coordinative versatility of FPhF to mu(2):eta(4):eta(3) coordination by the generation of K(2)(mu(2):eta(4):eta(3) FPhF)(2) units that exhibit eta(2)-arene interactions. Macromolecularly, the overlaying of these units affords a polymeric solvent-free structure that incites coordination of the FPhF ligands to metal atoms above and below the K(2)(FPhF)(2) plane. Overall, this generates a remarkable mu(4):eta(4):eta(3):eta(2):eta(1) amidinate binding mode that incorporates both bridging and terminal fluorine donors. Compounds 1-5 are the first non-chromium complexes of N,N'-di(ortho fluorophenyl)formamidinate. PMID- 12532291 TI - Diastereoselective hydroformylation of 2-substituted allylic o-DPPB-esters-on the origin of 1,2-asymmetric induction. AB - 2-Substituted secondary alcohol o-DPPB esters (o-DPPB=ortho diphenylphosphanylbenzoyl) have been prepared and their o-DPPB-directed diastereoselective hydroformylation examined. It was found that the diastereoselectivity increased as a function of the steric demand of the substituents both at the stereogenic center and in the alkene 2-position. Hydrolytic cleavage of the o-DPPB group afforded-via the lactols 29-the corresponding lactones 30, the relative configurations of the vicinal stereogenic centers of which were ascertainable by 2D-NOESY spectroscopy. In addition, a crystal structure analysis of the hydroformylation product 2 d provided further confirmation of the relative configuration. Replacement of the ester carbonyl group of the o-DPPB by a methylene unit resulted in significantly worse diastereoselectivity in the course of the hydroformylation (34-->35), which indicates a decisive role for the ester carbonyl function. All the experimental observations were combined in a model of the origin of the 1,2-asymmetric induction during the title reaction. The key feature is the consideration of diastereomeric trigonal-bipyramidal rhodium-hydrido-olefin complexes I and II, capable on the basis of the Hammond postulate of acting as good models for the transition states of the selectivity-determining hydrometalation step. Investigation of these complexes by force-field methods indicated good correlation between theoretically predicted and experimentally determined diastereoselectivities. PMID- 12532292 TI - Lanthanum(III)-catalyzed disproportionation of hydrogen peroxide: a heterogeneous generator of singlet molecular oxygen-1O2 (1Deltag)-in near-neutral aqueous and organic media for peroxidation of electron-rich substrates. AB - The decomposition of hydrogen peroxide into singlet molecular oxygen-(1)O(2) ((1)Delta(g))-in the presence of lanthanum(iii) salts was studied by monitoring its characteristic IR luminescence at 1270 nm. The process was found to be heterogeneously catalyzed by La(III), provided that the heterogeneous catalyst is generated in situ. The yield of (1)O(2) generation was assessed as 45+/-5 % both in water and in methanol. The pH-dependence on the rate of (1)O(2) generation corresponds to a bell-shaped curve from pH 4.5 to 13 with a maximum around pH 8. The study of the influence of H(2)O(2) showed that the formation of (1)O(2) begins as soon as one equivalent of H(2)O(2) is introduced. It then increases drastically up to two equivalents and more smoothly above. Unlike all other metal salt catalyst systems known to date for H(2)O(2) disproportionation, this chemical source of (1)O(2) is able to generate (1)O(2) not only in basic media, but also under neutral and slightly acidic conditions. In addition, this La-based catalyst system has a very low tendency to induce unwanted oxygenating side reactions, such as epoxidation of alkenes. These two characteristics of the heterogeneous lanthanum catalyst system allow non-photochemical (i.e., "dark") singlet oxygenation of substrate classes that cannot be peroxidized successfully with conventional molybdate catalysts, such as allylic alcohols and alkenyl amines. PMID- 12532293 TI - Flexible but with a defined turn-influence of the template on the binding properties of two-armed receptors. AB - Combinatorial binding studies revealed that the di(trans-4 aminoproline)diketopiperazine is an ideal template for two-armed receptors with highly selective binding properties towards peptides. It is not only superior to structurally very different diamines but also to the diastereomeric di(cis-4 aminoproline)diketopiperazine. These empiric results are rationalized by the analysis of the conformation of the diastereomeric diketopiperazines in the solid state, by X-ray crystal structure analysis, as well as by NMR studies in solution: to observe highly selective binding, the template needs to be not only conformationally rigid but it must have a specific turn geometry. The combination of combinatorial binding studies, X-ray crystal structure analysis, and NMR spectroscopy gave insight into why the trans,trans-diketopiperazine is a superior template compared to other diamines. Additionally, the results provide a guide for the rational design of two-armed receptors with good binding properties towards peptidic guests. PMID- 12532294 TI - 3,3'- and 4,4'-Dimethoxy-2,2'-bipyrroles: highly electron-rich model compounds for polypyrrole formation. AB - 3,3'-Dimethoxy-2,2'-bipyrrole (1) and 4,4'-dimethoxy-2,2'-bipyrrole (2) were obtained in short sequences and good yields from N-benzyl-3-hydroxypyrrole-2,4 dicarboxylic acid. The key intermediate leading to 1 is an N-benzyl-3 methoxypyrrole, which is dimerized by lithiation and oxidation with NiCl(2). The formation of 2 is achieved by a classical Ullmann coupling of diethyl 1-benzyl-2 bromo-4-methoxypyrrole-3,5-dicarboxylate. The N-benzyl protection groups of 1 and 2 are cleaved under reducing conditions with sodium in liquid ammonia. Both isomeric bipyrroles are extremely sensitive toward air. Compound 1 has a very low oxidation potential of 0.09 V against AgCl but film formation hardly occurs. On the other hand, compound 2 with a potential of 0.35 V readily forms stable polypyrrole films with anodic waves at -0.51 and -0.35 V and a cathodic wave at 0.77 V, the lowest potential ever observed for a p-doped polymer. PMID- 12532295 TI - Theoretical and photoluminescence studies on the d10-s2 AuI-TlI interaction in extended unsupported chains. AB - The reactions of solutions of TlPF(6) and OPPh(3) in tetrahydrofuran or acetone with NBu(4)[AuR(2)] (R=C(6)Cl(5), C(6)F(5)) gave the new complexes [Au(C(6)Cl(5))(2)](2)[Tl(OPPh(3))][Tl(OPPh(3))(L)] (L=THF (1), acetone (2)) and the previously reported [Tl(OPPh(3))(2)][Au(C(6)F(5))(2)] (3). The crystal structures of complexes 1 and 2 display extended unsupported chains with short intermolecular interactions between alternating gold(I) and thallium(I) centres. Moreover, the Tl(I) centres show two different types of geometrical environments, such as pseudotetrahedral and distorted trigonal-bipyramidal, due to the presence of solvent molecules that act as ligands in the solid-state structure. Quasirelativistic and nonrelativistic ab initio calculations were performed to study the nature of the intermetallic Au(I)-Tl(I) interactions and are consistent with the presence of a high ionic contribution (80 %) and dispersion-type (van der Waals) interaction with a charge-transfer contribution (20 %) when relativistic effects are taken into account. All complexes are luminescent in the solid state at room temperature and at 77 K. Complexes 1 and 2 show site selective excitation, probably due to the different environments around the Tl(I) centres. The DFT and time-dependent (TD)-DFT calculations are in agreement with the experimental excitation spectra for all complexes and confirm the site selective excitation behaviour as a function of the Tl(I) geometrical environment. PMID- 12532296 TI - A three-component reaction for diversity-oriented synthesis of polysubstituted piperidines: solution and solid-phase optimization of the first tandem aza[4+2]/allylboration. AB - This article describes the design and optimization of a simple three-component aza[4+2]/allylboration reaction to access polysubstituted alpha-hydroxyalkyl piperidines in a highly diastereocontrolled fashion from maleimides, 4 boronohydrazonodienes, and aldehydes. The aldehyde component does not interfere with the first aza[4+2] step, and it was found that this tandem reaction provides better yields of piperidine products 5 when carried out in one-pot. The required 4-borono-hydrazonodienes 1 are synthesized efficiently from the condensation of 3 boronoacrolein pinacol ester (4) with hydrazines. Overall, the three-component process using N-substituted maleimides as dienophiles produces four stereogenic centers and is quite general. It tolerates the use of a wide variety of aldehydes and hydrazine precursors with different electronic and steric characteristics. By allowing such a wide substrate scope and up to four elements of diversity, this reaction process is particularly well adapted towards applications in diversity oriented synthesis of polysubstituted piperidine derivatives. The suitability of the aza[4+2]/allylboration reaction for use in solid-phase chemistry was also demonstrated using a N-arylmaleidobenzoic acid functionalized resin. This novel multicomponent reaction thus offers a high level of stereocontrol and versatility in the preparation of densely functionalized nitrogen heterocycles. PMID- 12532297 TI - Synthesis, photophysical and electrochemical properties, and biological labeling studies of cyclometalated iridium(III) bis(pyridylbenzaldehyde) complexes: novel luminescent cross-linkers for biomolecules. AB - We report the synthesis, characterization, photophysical, and electrochemical properties of a series of luminescent cyclometalated iridium(III) complexes containing two aldehyde functional groups [Ir(pba)(2)(N-N)](PF(6)) (Hpba=4-(2 pyridyl)benzaldehyde; N-N=2,2'-bipyridine, bpy (1), 1,10-phenanthroline, phen (2), 3,4,7,8-tetramethyl-1,10-phenanthroline, 3,4,7,8-Me(4)-phen (3), 4,7 diphenyl-1,10-phenanthroline, 4,7-Ph(2)-phen (4)). The X-ray crystal structure of complex 1 has been investigated. Upon photoexcitation, complexes 1-4 exhibit intense and long-lived emission in fluid solutions at 298 K and in low temperature glass. The luminescence is assigned to a triplet intra-ligand ((3)IL) excited state associated with the pba(-) ligand, probably with mixing of some triplet metal-to-ligand charge-transfer ((3)MLCT) (dpi(Ir)-->pi*(pba(-))) character. Since each of these complexes possesses two aldehyde groups, which can react with the primary amine groups of biomolecules to form stable secondary amines after reductive amination, we have investigated the possibility of these complexes as novel luminescent cross-linkers for biological substrates. L-Alanine has been labeled with complexes 1-4 to give the luminescent bioconjugates 1 (Ala)(2)-4-(Ala)(2). These conjugates show strong photoluminescence with long emission lifetimes under ambient conditions. On the basis of the emission energy trend, the excited state of these luminescent bioconjugates is likely to bear a high parentage of (3)MLCT (dpi(Ir)-->pi*(N-N)) character. In addition, the glycoprotein avidin (Av) has also been conjugated with complexes 1-4 to give the bioconjugates 1-Av-4-Av. Upon photoexcitation, these bioconjugates also display intense and long-lived (3)MLCT (dpi(Ir)-->pi*(N-N)) emission in aqueous buffer at 298 K. Furthermore, a heterogeneous competitive assay for biotin has been developed using 2-Av and biotinylated microspheres. We have shown that complexes 1-4 represent a new class of multicolor luminescent cross-linkers for biomolecular species. PMID- 12532298 TI - A comprehensive theoretical study on the reactions of Sc+ with CnH2n+2 (n=1-3): structure, mechanism, and potential-energy surface. AB - The reactions of Sc(+)((3)D) with methane, ethane, and propane in the gas phase were studied theoretically by density functional theory. The potential energy surfaces corresponding to [Sc, C(n), H(2n+2)](+) (n=1-3) were examined in detail at the B3LYP/6-311++G(3df, 3pd)//B3LYP/6-311+G(d,p) level of theory. The performance of this theoretical method was calibrated with respect to the available thermochemical data. Calculations indicated that the reactions of Sc(+) with alkanes are multichannel processes which involve two general mechanisms: an addition-elimination mechanism, which is in good agreement with the general mechanism proposed from earlier experiments, and a concerted mechanism, which is presented for the first time in this work. The addition-elimination reactions are favorable at low energy, and the concerted reactions could be alternative pathways at high energy. In most cases, the energetic bottleneck in the addition elimination mechanism is the initial C--C or C--H activation. The loss of CH(4) and/or C(2)H(6) from Sc(+)+C(n)H(2n+2) (n=2, 3) can proceed along both the initial C--C activation branch and the Cbond;H activation branch. The loss of H(2) from Sc(+)+C(n)H(2n+2) (n=2, 3) can proceed not only by 1,2-H(2) and/or 1,3 H(2) elimination, but also by 1,1-H(2) elimination. The reactivity of Sc(+) with alkanes is compared with those reported earlier for the reactions of the late first-row transition-metal ions with alkanes. PMID- 12532300 TI - Synthesis and structural analysis of the polymetallated alkali calixarenes [M4(p tert-butylcalix[4]arene-4H)(thf)x]2.n THF (M=Li, K; n=6 or 1; x=4 or 5) and [Li2(p-tert-butylcalix[4]arene-2H)(H2O)(mu-H2O)(thf)].3 THF. AB - To study the structures and reactivities of alkali metallated intermediates of calix[4]arenes, three compounds were isolated: [Li(4)(p-tert-butylcalix[4]arene 4H)(thf)(4)](2).6 THF (1), [Li(2)(p-tert-butylcalix[4]arene-2H)(H(2)O)(mu H(2)O)(thf)].3 THF (2), and [K(4)(p-tert-butylcalix[4]arene-4H)(thf)(5)](2).THF (3). The structure of 1 is shown to be dependent on the coordinating solvent. Partial hydrolysis of 1 leads to the formation of 2. The potassium compound 3 features a different structure to that of 1, due to a higher coordination number as well as stronger cation-pi-bonding interactions. PMID- 12532299 TI - Substituent effects on O--H bond dissociation enthalpies and ionization potentials of catechols: a DFT study and its implications in the rational design of phenolic antioxidants and elucidation of structure-activity relationships for flavonoid antioxidants. AB - Density functional theory (DFT) on B3LYP/6-31G(d,p) level was employed to investigate the substituent effects on O--H bond dissociation enthalpies (BDEs) and ionization potentials (IPs) of catechols. It was revealed that the ortho hydroxyl of catechol was effective for the reduction of the O--H BDE; however, the group had little influence on the IP. The para substituent effects upon O--H BDEs and IPs for catechols were roughly the same as those for monophenols, and this gave the catechol moiety more potential than monophenol to be used as a lead compound in rational design of phenolic antioxidants. In addition, the 1,4-pyrone effects on O--H BDEs of catecholic rings A or B of flavonoids were also investigated. Although 1,4-pyrone extended the conjugation system of flavonoids, it was not beneficial to reduce the O--H BDE as a result of its electron withdrawing property. Thus, 1,4-pyrone was unlikely to be favorable to enhance the H-abstraction activity of flavonoids. PMID- 12532301 TI - The stabilization of monomeric parent compounds of phosphanyl- and arsanylboranes. AB - The structures of the parent compounds of phosphanyl- and arsanylboranes, H(2)BPH(2) and H(2)BAsH(2), were calculated by DFT-B3LYP methods. Such compounds have not previously been obtained preparatively. By applying the concept of Lewis acid/base stabilisation, [(CO)(5)W(H(2)EBH(2).NMe(3))] (E=P (3), As (4)) derivatives have been synthesised by the metathesis reactions between Li[(CO)(5)WEH(2)] and ClH(2)BNMe(3) (E=P, As). Comprehensive thermodynamic studies on these systems verify the high stability of the Lewis acid/base stabilised complexes. Unexpected based on the thermodynamic calculations, UV radiation of the phosphanylborane 3 leads to the dinuclear phosphanido-bridged complex [(CO)(8)W(2)(mu-PHBH(2).NMe(3))(2)] (5) by H(2) and CO elimination. PMID- 12532302 TI - Side-on bridging coordination of N2: spectroscopic characterization of the planar Zr2N2 core and theoretical investigation of its butterfly distortion. AB - The vibrational and electronic structure of the side-on N(2)-bridged Zr complex [((P(2)N(2))Zr)(2)(mu-eta(2):eta(2)-N(2))] (P(2)N(2)=PhP(CH(2)SiMe(2)NSiMe(2)CH(2))(2)PPh) were analyzed. The vibrational characterization of the planar Zr(2)N(2) core was based on resonance Raman and infrared spectroscopy. In the Raman spectrum, the Nbond;N stretching band is found at 775 cm(-1) with an isotope shift of 22 cm(-1). Due to its appearance in many overtones and combination modes, the metal-metal stretch is assigned to the peak at 295 cm(-1). The two ungerade modes of the Zr(2)N(2) core were identified in the infrared spectrum. Based on these four vibrations of the Zr(2)N(2) unit, a quantum chemical assisted normal coordinate analysis (QCA-NCA) was performed. The force constants for the N--N and Zr--N bonds were calculated to be 1.53 and 2.58 mdyn A(-1), respectively. The butterfly distortion of the Zr(2)N(2) unit obtained in DFT geometry optimizations of planar side-on N(2)-bridged Zr complexes was analyzed in more detail. It was found that on bending of the Zr(2)N(2) core, the lone pairs of the axial amide ligands are rotated by 90 degrees. The bent Zr(2)N(2) unit is 11 kcal mol(-1) lower in energy than the planar core due to a more uniform distribution of electron density between the metal atoms and N(2) and delocalization of electron density from the amide ligands to the Zr(2)N(2) unit. The spectroscopic implications of this distortion are analyzed. PMID- 12532303 TI - Reagent-controlled stereoselectivity in titanocene-catalyzed epoxide openings: reductions and intermolecular additions to alpha,beta-unsaturated carbonyl compounds. AB - The generation and addition reactions of metal bound radicals derived from normal and meso epoxides by electron transfer from titanocene(III) reagents is described. The control of enantioselectivity and diastereoselectivity of these transformations is investigated by variation of the ligands of the metal complex. The reaction can lead to unprecedented and highly selective reactions, in which synthetically useful alcohols may be prepared. The synthesis presented also circumvents the use of toxic metals. Another advantage is that there is no loss of two functional groups as usually observed in reductive radical chain reactions. PMID- 12532304 TI - Dynamic chirality: keen selection in the face of stereochemical diversity in mechanically bonded compounds. AB - The template-directed syntheses, employing bisparaphenylene-[34]crown-10 (BPP34C10), 1,5-dinaphthoparaphenylene-[36]crown-10 (1/5NPPP36C10), and 1,5 dinaphtho-[38]crown-10 (1/5DNP38C10) as templates, of three [2]catenanes, whereby one of the two bipyridinium units in cyclobis(paraquat-p-phenylene) is replaced by a bipicolinium unit, are described. The crude reaction mixtures comprising the [2]catenanes all contain slightly more of the homologous [3]catenanes, wherein a "dimeric" octacationic cyclophane has the crown ether macrocycles encircling the alternating bipyridinium units with the bipicolinium units completely unfettered. X-ray crystallography, performed on all three [2]catenanes and two of the three [3]catenanes reveals co-conformational and stereochemical preferences that are stark and pronounced. Both the [3]catenanes crystallize as mixtures of diastereoisomers on account of the axial chirality associated with the picolinium units in the solid state. Dynamic (1)H NMR spectroscopy is employed to probe in solution the relative energy barriers for rotations by the phenylene and pyridinium rings in the tetracationic cyclophane component of the [2]catenanes. Where there are co-conformational changes that are stereochemically "allowed", crown ether circumrotation and rocking processes are also investigated for the relative rates of their occurrence. The outcome is one whereby the three [2]catenanes containing BPP34C10, 1/5NPPP36C10, and 1/5DNP38C10 exist as one major enantiomeric pair of diastereoisomers amongst two, four, and eight diastereoisomeric pairs of enantiomers, respectively. The diastereoisomerism is a consequence of the presence of axial chirality together with helical and/or planar chirality in the same interlocked molecule. These [2]catenanes constitute a rich reserve of new stereochemical types that might be tapped for their switching and mechanical properties. PMID- 12532305 TI - Synthesis of an amino-functionalized model of the Fe-only hydrogenase active site. AB - A dinuclear 2Fe2S mimic 6 of the active site of the Fe-only hydrogenases has been synthesized. Complex 6 contains a free amino group which enables linkage to a protein backbone or to a redox active species for the study of electron transfer processes in proteins or in supramolecular systems. The structures of the complex 6 and its Boc-protected precursor 5 could be verified by X-ray crystallography. PMID- 12532306 TI - Nitrogen photofixation on nanostructured iron titanate films. AB - A nanostructured iron titanate thin film has been prepared by a sol-gel method from iron(III) chloride and titanium tetraisopropylate. Energy-dispersive X-ray analysis and Mossbauer spectroscopy suggest the presence of a Fe(2)Ti(2)O(7) phase, which was previously obtained as an intermediary phase upon heating ilmenite. In the presence of ethanol or humic acids and traces of oxygen, the novel film photocatalyzes the fixation of dinitrogen to ammonia (17 microM) and nitrate (45 microM). In the first observable reaction step, hydrazine is produced and then undergoes further photoreduction to ammonia. Oxidation of the latter by oxygen affords nitrate as the final product. Since the reaction occurs also in air and with visible light (lambda>455 nm), and since the iron titanate phase may be formed by the weathering of ilmenite minerals, it may be a model for mutual nonenzymatic nitrogen fixation in nature. PMID- 12532307 TI - On the cavitation energy of water. AB - Free-energy-perturbation theory from molecular dynamics calculations has been used to obtain the DeltaG of adjoining cavities' formation in water. The DeltaGs for systems with three, five and seven cavities are compared with that of a single cavity of the same volume, and found to be in good agreement. The conditions under which the analytical formulation of the energy of cavity formation proposed by Pierotti holds are discussed. The data for a single cavity have been tabulated and can lend themselves to a simple numerical implementation in standard quantum chemical packages, which can be used when high accuracy for DeltaG(cav) is required. PMID- 12532308 TI - Sequestered alkyllithiums: why phenyllithium alone is suitable for betaine-ylide generation. AB - The key step in the trans-selective modification of the Wittig reaction is the alpha-lithiation of the lithium bromide coordinated ylide-aldehyde adduct (the so called "P-betaine"). Only phenyllithium effects this deprotonation rapidly and cleanly. Alkyllithiums (in particular, butyl-, sec-butyl-, and tert-butyllithium) react only sluggishly and incompletely, being tied up in very stable mixed aggregates with the lithium alkoxide part of the betaines. PMID- 12532309 TI - Vibronic coupling in molecules and in solids. AB - We utilize the experience gained in our previous studies on the "chemistry of vibronic coupling" in simple homonuclear and heteronuclear molecules to begin assembling theoretical guidelines for the construction of potentially superconducting solids exhibiting large electron-phonon coupling. For this purpose we analyze similarities between vibronic coupling in isolated molecules and in extended solids. In particular, we study vibronic coupling along the antisymmetric stretch coordinate (Q(as)) in linear symmetric AAA molecules, and along the optical phonon "pairing" mode coordinate (Q(opt)) in corresponding one dimensional [A]( infinity ) chains built of equidistant A atoms. This is done for a broad range of chemical elements (A). The following similarities between vibronic coupling in molecules and phonon coupling in solids emerge from our calculations: 1) The HOMO/LUMO electronic energy gap in an AAA molecule increases along Q(as), and the highest occupied crystal orbital/lowest unoccupied crystal orbital gap in [A]( infinity ) chain increases along Q(opt). 2) The maximum vibronic instability is invariably obtained for a half-filled, singly occupied molecular orbital in AAA molecules, and for a corresponding half-filled band in [A]( infinity ) chains. 3) The vibronic stability of an AAA molecule increases with a decrease of the AA bond length, as does the vibronic stability of [A]( infinity ) chains (external pressure may lead to a reversal of a Peierls distortion). 4) The high degree of s-p mixing and ionic/covalent forbidden curve crossing dramatically enhance the vibronic instability of both AAA molecules and [A]( infinity ) chains. We also introduce one quantitative relationship: The parameter log(R) (where R is molar refractivity, a parameter used by Herzfeld to prescribe the conditions for the metallization of the elements) correlates with a parameter f(AA) (defined as twice the electronegativity of A, divided by the equilibrium AA bond length), used by two of us previously to describe vibronic coupling in AAA molecules for a broad range of elements (A=halogen, H, or an alkali metal). We hope to illustrate that key chemical aspects of vibronic coupling in simple molecules may thus be profitably transferred to corresponding materials in the solid state. PMID- 12532311 TI - Subjective assessment of the effects of CNS-active drugs on sleep by the Leeds sleep evaluation questionnaire: a review. AB - The Leeds sleep evaluation questionnaire (LSEQ) comprises ten self-rating 100 mm line analogue questions concerned with sleep and early morning behaviour. A literature search identified 83 studies in peer-reviewed journals that reported the use of the LSEQ for psychopharmacological investigations of drug effects on self-reported aspects of sleep. High internal consistency and reliability of the questionnaire have been demonstrated. Findings from studies involving a variety of psychoactive agents indicated that the LSEQ was able to quantify subjective impressions of sleep and waking and the effects of drugs in healthy volunteers, depressed and insomnia patients. In accordance with their known activity profile nocturnal administration of sedative hypnotic agents and antihistamines induced dose-related improvements in self-reported ease of getting to sleep, and quality of sleep but a decrease in alertness and behavioural integrity the following morning. Psychostimulants, on the other hand, impaired subjective ratings of sleep and increased early morning alertness. Antidepressants and certain anxiolytic agents improved both self-reported sleep aspects and early morning alertness. Treatment effects measured by the LSEQ corresponded to those measured for the same drugs by other assessment methods. These data indicate that the LSEQ is a robust and reliable instrument for psychopharmacological evaluations. Self evaluations of sleep, as obtained by the LSEQ, can therefore provide consistent and meaningful measures for estimating the effectiveness of sleep modulators and sedative-hypnotic drugs. PMID- 12532312 TI - Quantifying subjective assessment of sleep and life-quality in antidepressant treated depressed patients. AB - This study sought to establish a method of quantifying subjective perceptions of sleep against perceptions of life-quality and mood, using amended versions of the Pittsburgh sleep diary (PghSD) and quality of life of insomniacs (QOLI) questionnaire. Diaries and questionnaires were self-completed in participants' homes. Outpatients with a DSM-IV diagnosis of major depressive disorder were compared with a healthy control group (with no history, or family history, of depression). Poorer sleepers, as determined by the sleep diary, were significantly more likely to report poorer life-quality and mood perceptions on the subsequent questionnaire. Furthermore, the depressed group reported significantly poorer perceptions of sleep quality and poorer perceptions of life quality and mood than the control group, even though estimates of sleep disturbance were similar. This may indicate that depressed individuals experience more 'sleep distress' than healthy individuals. These results confirm the extent of subjectively reported sleep disruption in depression and demonstrate the merit of combining the amended PghSD and QOLI to quantify sleep perceptions. PMID- 12532313 TI - Zopiclone and zaleplon vs benzodiazepines in the treatment of insomnia: Canadian consensus statement. PMID- 12532314 TI - Actigraphy in human psychopharmacology: a review. AB - Over the last 25 years wrist actigraphy has predominantly been used in sleep research and chronobiology where the technique has proved useful in the measurement of sleep and the characterisation of the sleep/wake cycle. Whilst there are a large number of studies published that have used actigraphy, there have only been relatively few studies that have shown that the technique is capable of measuring drug-induced changes in nocturnal and/or diurnal behaviour. Thus the use of actigraphy to measure drug effects in clinical trials has up till now remained on the periphery of psychopharmacology. However this may be because of the disparate nature of the studies published rather than lack of usefulness. This paper reviews the relevant literature and in doing so concludes that there is a convincing body of evidence for the utility of actigraphy as a technique in human psychopharmacology. PMID- 12532315 TI - Cognitive effects of long-term benzodiazepine use in older adults. AB - This study examined the potential for cognitive morbidity associated with the long-term use of benzodiazepine (BZ) sedative-hypnotics in a sample of healthy older adults. Tests of memory, attention and processing speed were conducted prior to and 1 month after drug discontinuation for 25 BZ-users and at similar intervals for 26 healthy control subjects. After controlling for differences in affective status between BZ-users and controls, there were no significant group differences in cognitive performance. However, BZ-users showed greater gains on tests of attention and speed of processing at repeat testing compared with controls this improvement was not attributable to a change in affective status. These findings suggest that there may be subtle and reversible effects of long term BZ use on speed-dependent tasks in older adults. However, the magnitude of these effects is quite small and may be of little clinical significance in the healthy elderly. PMID- 12532316 TI - Drug induced nightmares--an etiology based review. AB - OBJECTIVE: Recent clinical trials have included patient complaints of nightmares as a category of reportable medication side effects. This study integrates that data into current experimental and theoretical research of drug effects that may alter dreaming and nightmares. The objective is to provide a clinical and theoretical framework useful in categorizing the potential and reported drug effects on nightmares. METHODOLOGY: This study reviews case reports and clinical trials that have reported nightmares or alterations in dreaming occurring secondary to medication usage. These data are analysed as to the probability of the drug/nightmare association, and integrated into current electrophysiological and neurochemical theories of dreaming and nightmares. RESULTS: Pharmacological agents affecting the neurotransmitters norepinephrine, serotonin and dopamine are clearly associated with patient reports of nightmares. Agents affecting immunological response to infectious disease are likely to induce nightmares in some patients. A possible association exists between reports of nightmares and agents affecting the neurotransmitters acetylcholine, GABA and histamine, as well as for some anesthetics, antipsychotics and antiepileptic agents. CONCLUSION: By utilizing our current experimental and theoretical knowledge base, the potential etiology of a majority of reported drug effects on nightmares can be classified. These data support current neurochemical theories of dreaming, as well as suggesting that the biochemical basis for dreaming and nightmare induction may be more complex than generally suggested. PMID- 12532317 TI - Are there gender differences in the prescribing of hypnotic medications for insomnia? AB - Gender differences in the prescribing patterns of general classes of medications for insomnia were examined. The classes of medications included: zopiclone, antidepressants, benzodiazepines, antihistamines and no medication. The sample comprised a sub-set of respondents from 2620 questionnaires of the Canadian Multicentre Sleep Database. Respondents for this database were contacted through physicians, announcements in the media and local pharmacies. The results indicated that gender alone was not associated with differential prescribing for insomnia, nor was gender associated with patterns of medication use such as frequency of taking medication, length of use, taking more or less medication than prescribed or attempts to stop taking medication. Demographic factors were included in the analysis and age and marital status were associated with different prescribing patterns for men and women with insomnia. It is possible that physicians refer to stereotypic expectations when prescribing hypnotics. PMID- 12532318 TI - Adverse effects of temazepam in older adults with chronic insomnia. AB - BACKGROUND: Benzodiazepine-hypnotics are frequently used for treating insomnia in older adults; however, there is little information about adverse effects associated with their usage over several weeks, particularly in this segment of the population. OBJECTIVE: This study reports on the incidence of adverse effects of temazepam in older adults with chronic insomnia and examines whether the addition of cognitive-behaviour therapy (CBT) is associated with less drug used and fewer adverse effects. METHOD: Sixty patients with chronic and primary insomnia were randomized to temazepam (n = 20), placebo (n = 20) or temazepam plus CBT (n = 20). Data from the physicians' weekly assessments and patients' sleep diaries were used to evaluate adverse effects, the dose (7.5-30 mg) at which they occurred, and drug use patterns over the 8-week course of treatment. RESULTS: The incidence of adverse effects was infrequent, as shown by the low percentages of complaints reported by patients in the temazepam (7.8%), placebo (10.8%) and combination groups (8.3%). The severity of adverse events was mild and decreased over the course of treatment. The maximum dose was reached by 10 patients receiving temazepam, 14 placebo and 7 in the combined treatment. The average nightly dosage used was 20 mg for both the temazepam and placebo groups and 16 mg for the combined condition. Patients receiving temazepam plus CBT used less drugs, with approximately the same incidence of adverse effects. CONCLUSION: Temazepam is a safe hypnotic for use by older adults over an 8-week treatment period. There are few adverse effects and behavioural tolerance to those effects develop over time. The addition of cognitive-behavioural intervention reduced both the amount of medication used and the incidence of adverse effects, with comparable sleep improvements. PMID- 12532319 TI - Simulated microgravity suppresses osteoblast phenotype, Runx2 levels and AP-1 transactivation. AB - Conditions of disuse such as bed rest, space flight, and immobilization result in decreased mechanical loading of bone, which is associated with reduced bone mineral density and increased fracture risk. Mechanisms involved in this process are not well understood but involve the suppression of osteoblast function. To elucidate the influence of mechanical unloading on osteoblasts, a rotating wall vessel (RWV) was employed as a ground based model of simulated microgravity. Mouse MC3T3-E1 osteoblasts were grown on microcarrier beads for 14 days and then placed in the RWV for 24 h. Consistent with decreased bone formation during actual spaceflight conditions, alkaline phosphatase and osteocalcin expression were decreased by 80 and 50%, respectively. In addition, runx2 expression and AP 1 transactivation, key regulators of osteoblast differentiation and bone formation, were reduced by more than 60%. This finding suggests that simulated microgravity could promote dedifferentiation and/or transdifferentiation to alternative cell types; however, markers of adipocyte, chondrocyte, and myocyte lineages were not induced by RWV exposure. Taken together, our results indicate that simulated microgravity may suppress osteoblast differentiation through decreased runx2 and AP-1 activities. PMID- 12532320 TI - Membrane receptors for steroid hormones: signal transduction and physiological significance. AB - Membrane receptors for steroid hormones affect signaling pathways that modulate nuclear function, influence neuronal activity, ion flow, and the circulatory system. Indeed, 'new' steroid hormones have been identified by their interaction with membrane-initiated signaling systems. A brief summary of the FASEB Summer Research Conference devoted to these topics is presented in this mini-review. In addition, attendees of the meeting propose introduction of the following terminology: membrane-initiated steroid signaling (MISS) and nuclear-initiated steroid signaling (NISS) to replace more inaccurate terms in current use. PMID- 12532321 TI - Regulation of the osteoblast-specific transcription factor, Runx2: responsiveness to multiple signal transduction pathways. AB - The Cbfa1/Runx2 is an important transcription factor necessary for osteoblast differentiation and bone formation. However, the signaling pathways regulating Runx2 activity are just beginning to be understood. Inconsistencies between Runx2 mRNA or protein levels and its transcriptional activity suggests that posttranslational modification and/or protein-protein interactions may regulate this factor. Runx2 can be phosphorylated and activated by the mitogen-activated protein kinase (MAPK) pathway. This pathway can be stimulated by a variety of signals including those initiated by extracellular matrix (ECM), osteogenic growth factors like bone morphogenic proteins (BMPs) and fibroblast growth factor 2 (FGF-2), mechanical loading and hormones such as parathyroid hormone (PTH). Protein kinase A (PKA) may also phosphorylate/activate Runx2 under certain conditions. In addition, Runx2 activity is enhanced by protein-protein interactions as are seen with PTH-induced Runx2/AP-1 and BMP-mediated Runx2/Smads interactions. Mechanisms for interaction with Runx2 are complex including binding of distinct components such as AP-1 factors and Smads proteins to separate DNA regions in target gene promoters and direct physical interactions between Runx2 and AP-1/Smad factors. Post-translational modifications such as phosphorylation may influence interactions between Runx2 and other nuclear factors. These findings suggest that Runx2 plays a central role in coordinating multiple signals involved in osteoblast differentiation. PMID- 12532322 TI - Nuclear lipids: new functions for old molecules? AB - It is becoming increasingly evident that stimulation of nuclear lipid metabolism plays a central role in many signal transduction pathways that ultimately result in various cell responses including proliferation and differentiation. Nuclear lipid metabolism seems to be at least as complex as that existing at the plasma membrane. However, a distinctive feature of nuclear lipid biochemical pathways is their operational independence from their cell periphery counterparts. Although initially it was thought that nuclear lipids would serve as a source for second messengers, recent evidence points to the likelihood that lipids present in the nucleus also fulfil other roles. The aim of this review is to highlight the most intriguing advances made in the field over the last year, such as the production of new probes for the in situ mapping of nuclear phosphoinositides, the identification of two sources for nuclear diacylglycerol production, the emerging details about the peculiar regulation of nuclear phosphoinositide synthesizing enzymes, and the distinct possibility that nuclear lipids are involved in processes such as chromatin organization and pre-mRNA splicing. PMID- 12532323 TI - Prospects for cannabinoids as anti-inflammatory agents. AB - The marijuana plant (Cannabis sativa) and preparations derived from it have been used for medicinal purposes for thousands of years. It is likely that the therapeutic benefits of smoked marijuana are due to some combination of its more than 60 cannabinoids and 200-250 non-cannabinoid constituents. Several marijuana constituents, the carboxylic acid metabolites of tetrahydrocannabinol, and synthetic analogs are free of cannabimimetic central nervous system activity, do not produce behavioral changes in humans, and are effective antiinflammatory and analgesic agents. One cannabinoid acid in particular, ajulemic acid, has been studied extensively in in vitro systems and animal models of inflammation and immune responses. This commentary reviews a portion of the work done by investigators interested in separating the medicinal properties of marijuana from its psychoactive effects. Understanding the mechanisms of the therapeutic effects of nonpsychoactive cannabinoids should lead to development of safe effective treatment for several diseases, and may render moot the debate about "medical marijuana". PMID- 12532324 TI - Gene therapy and tissue engineering in repair of the musculoskeletal system. AB - Historically, surgeons have sought and used different procedures in order to augment the repair of various skeletal tissues. Now, with the completion of the Human Genome Project, many researchers have turned to gene therapy as a means to aid various ailments. In the orthopedic field, many strides have been made toward using gene therapy and tissue engineering in a clinical setting. In this review, several studies are outlined in different areas that gene therapy has or will influence orthopedic surgery. Gene therapy and tissue engineering can aid in fracture healing and spinal fusions by inducing bone formation, ligamentous repairs by increasing the production of connective tissue fibers, intervertebral disc disease by creating potential replacements, and articular cartilage repairs by providing means to improve cartilage. As we continue to see great contributions, such as the few mentioned here, this field will continue to mature and develop. PMID- 12532325 TI - Activation of multiple caspases and modification of cell surface fas (CD95) in proteasome inhibitor-induced apoptosis of rat natural killer cells. AB - The proteasome is a multi-subunit protease complex that is involved in intracellular protein degradation in eukaryotes. Previously, we have reported that selective, synthetic chymotryptic proteasome inhibitors inhibit A-NK cell mediated cytotoxicity by approximately 50%; however, the exact role of the proteasome in NK cell-mediated cytotoxicity remains unknown. Herein, we report that proteasome inhibitors, MG115 and MG132, decreased the proteasome chymotrypsin-like activity in the rat natural killer cell line RNK16 by 85% at a concentration of 5 microM. The viability of RNK16 cells was also reduced in the presence of these inhibitors. Both inhibitors induced the apoptosis of RNK16 cells, as shown by DNA fragmentation, caspase-3 activation and the appearance of sub-G-cell populations. An increase in the fraction of apoptotic cells was observed in a dose- and time-dependent manner in our studies. In addition, the activity of caspase-1, -2, -6, -7, -8, and -9, was increased following the treatment of RNK16 cells with these inhibitors. Further investigation revealed that the expression of Fas (CD95) protein on the RNK16 cell surface was increased after the treatment by MG115 or MG132, indicating that apoptosis induced by proteasome inhibitors in RNK16 cells might be mediated through the Fas (CD95) mediated death pathway as well. Our studies indicate, for the first time, that proteasomal chymotryptic inhibitors can reduce natural killer cell viability and therefore indirectly inhibit cell-mediated cytotoxicity via the apoptosis inducing properties of these agents. PMID- 12532326 TI - IRES-dependent translational control of Cbfa1/Runx2 expression. AB - The P1 and P2 promoters of the Cbfa1/Runx2 gene produce Type I and II mRNAs with distinct complex 5'-untranslated regions, respectively designated UTR1 and UTR2. To evaluate whether the 5'-UTRs impart different translational efficiencies to the two isoforms, we created SV40 promoter-UTR-luciferase reporter (luc) constructs in which the translational potential of the 5'-UTR regions was assessed indirectly by measurement of luciferase activity in transfected cell lines in vitro. In MC3T3-E1 pre-osteoblasts, UTR2 was translated approximately twice as efficiently as the splice variants of UTR1, whereas translation of unspliced UTR1 was repressed. To determine if the UTRs conferred internal ribosome entry site (IRES)-dependent translation, we tested bicistronic SV40 promoter-Rluc-UTR-Fluc constructs in which Fluc is expressed only if the intercistronic UTR permits IRES-mediated translation. Transfection of bicistronic constructs into MC3T3-E1 osteoblasts demonstrated that both UTR2 and the spliced forms of UTR1 possess IRES activity. Similar to other cellular IRESs, activity increased with genotoxic stress induced by mitomycin C. In addition, we observed an osteoblastic maturation-dependent increase in IRES-mediated translation of both UTR2 and the spliced forms of UTR1. These findings suggest that Cbfa1 UTRs have IRES-dependent translational activities that may permit continued Cbfa1 expression under conditions that are not optimal for cap-dependent translation. PMID- 12532327 TI - Differential regulation of cellular adhesion and migration by recombinant laminin 5 forms with partial deletion or mutation within the G3 domain of alpha3 chain. AB - The basement membrane protein laminin-5 promotes cell adhesion and migration. The carboxyl-terminal G3 domain in the alpha3 chain is essential for the unique activity of laminin-5. To investigate the function of the G3 domain, we prepared various recombinant laminin-5 forms with a partially deleted or mutated G3 domain. The deletion of the carboxyl-terminal 28 amino acids (region III) markedly decreased the cell adhesion activity with a slight loss of the cell motility activity toward BRL and EJ-1 cells. This change was attributed to the loss of Lys-Arg-Asp sequence. Further deletion of 83 amino acids (region II) led to almost complete loss of the cell motility activity. All charged amino acid residues tested in this region were not responsible for the activity loss. These results suggest that the G3 domain contains two distinct regions that differently regulate cell adhesion and migration. Analysis of laminin-5 receptors showed that integrins alpha3beta1, alpha6beta1, and alpha6beta4 had different but synergistic effects on cell adhesion and migration on laminin-5. However, the structural change of the G3 domain appeared not to change integrin specificity. The present study demonstrates that the G3 domain in laminin-5 plays a central role to produce different biological effects on cells. PMID- 12532328 TI - Myosin heavy chain kinase B participates in the regulation of myosin assembly into the cytoskeleton. AB - Myosin II plays critical roles in events such as cytokinesis, chemotactic migration, and morphological changes during multicellular development. The amoeba Dictyostelium discoideum provides a simple system for the study of this contractile protein. In this system, myosin II filament assembly is regulated by myosin heavy chain (MHC) phosphorylation in the tail region of the molecule. Earlier studies identified an alpha-kinase, MHC kinase A (MHCK A), which phosphorylates three mapped threonine residues in the myosin tail, driving myosin disassembly. Using molecular and genomic approaches, we have identified a series of related kinases in Dictyostelium. The enzyme MHCK B shares with MHCK A a domain organization that includes a highly novel catalytic domain coupled to a carboxyl-terminal WD repeat domain. We have engineered, expressed, and purified a FLAG-tagged version of the novel kinase. In the present study, we report detailed biochemical and cellular studies documenting that MHCK B plays a physiological role in the control of Dictyostelium myosin II assembly and disassembly during the vegetative life of Dictyostelium amoebae. The presented data supports a model of multiple related MHCKs in this system, with different regulatory mechanisms and pathways controlling each enzyme. PMID- 12532329 TI - Changes of sphingolipid species in the phenotype conversion from myofibroblasts to lipocytes in hepatic stellate cells. AB - Sphingolipids play a relevant role in cell-cell interaction, communication, and migration. We studied the sphingolipid content in the murine hepatic stellate cell line GRX, which expresses the myofibroblast phenotype, and can be induced in vitro to display the fat-storing phenotype. Lipid modifications along this induction were investigated by labeling sphingolipids with [(14)C]galactose, [(14)C]serine, or [(14)C]choline, and determination of fatty acid composition of sphingomyelin. The total ganglioside content and the GM2 synthase activity were lower in myofibroblasts. Both phenotypes presented similar gangliosides of the a pathway: GM2, GM1, and GD1a as well as their precursor GM3. Sphingomyelin and all the gangliosides were expressed as doublets; the upper/lower band ratio increased in lipocytes, containing more long-chain fatty acids in retinol-induced lipocytes as compared to the insulin/indomethacin induced ones. Time-course experiments indicated a transfer of metabolic precursors from phosphatidylcholine to sphingomyelin in the two phenotypes. Taken together, these results indicate that myofibroblast and lipocytes can use distinct ceramide pools for sphingolipid synthesis. Differential ganglioside expression and presence of the long-chain saturated fatty acids suggested that they may participate in formation of distinct membrane microdomains or rafts with specific functions on the two phenotypes of GRX-cells. PMID- 12532330 TI - Raf-independent and MEKK1-dependent activation of NF-kappaB by hydrogen peroxide in 70Z/3 pre-B lymphocyte tumor cells. AB - We have previously demonstrated that hydrogen peroxide (H(2)O(2)) treatment of murine 70Z/3 pre-B lymphocytes inhibits the immune response to lipopolysaccharide by attenuating signaling through c-Jun N-terminal kinase (JNK) activation. In the present study, we further examined the signaling intermediates responsible for immunosuppression by H(2)O(2), focusing on NF-kappaB, a dimeric transcription factor whose activation is implicated in a number of immune response. Treatment of 70Z/3 pre-B cells with H(2)O(2) caused activation of NF-kappaB in the nuclei by detection of NF-kappaB specific DNA binding, concomitant with phosphorylation of IkappaBalpha. H(2)O(2) stimulation of NF-kappaB occurred within 20 min of treatment, reached maximum level at 60 min, and sustained for 2 h or more. Especially, MEK1 may contribute to H(2)O(2)-induced NF-kappaB activation as shown in the inhibition of NF-kappaB binding activity by the MEK1 inhibitor, PD 98059, and H(2)O(2)-induced MEK1 activation. However, H(2)O(2) exhibited no effect on the activity of Raf-1 kinase, which was an upstream activator of MEK1. Furthermore, B-58l and alpha-hydroxyfarnesylphosphonic acid, two inhibitors of Ras, did not block NF-kappaB activation. In addition, the transient transfection of a dominant negative Ras (RasN17) construct showed a negligible inhibitory effect on the activation of NF-kappaB by H(2)O(2). Instead, treatment of 70Z/3 cells with H(2)O(2) resulted in the activation of MAPK kinase kinase 1 (MEKK1) as well as JNK. Therefore, our data suggest that H(2)O(2) regulates the activity of NF-kappaB by MEK1 activation through MEKK1-dependent but Ras/Raf-independent mechanism. PMID- 12532331 TI - The E2F-1 transcription factor is negatively regulated by its interaction with the MDMX protein. AB - Several proteins with important roles in oncogenesis have been shown to regulate the function of the E2F-1 transcription factor, which is known to activate the expression of genes required for proliferation and apoptosis. Here we identify the MDMX oncoprotein as an E2F-1-binding factor, from a yeast-two hybrid screen using a portion of the E2F-1 protein as "bait." We demonstrate that the region within MDMX needed for the E2F-1:MDMX interaction is located in the central part of the protein, C-terminal of the p53-binding domain. The region within E2F-1 needed for this association is adjacent to the DNA binding domain. Further, when expressed in vivo or in vitro the MDMX protein migrates as two isoforms on SDS PAGE, the faster migrating isoform having the stronger affinity for the E2F-1 proteins. It appears that this interaction reduces the ability of E2F-1 to bind DNA. Expression of MDMX along with E2F-1 and Dp-1 in Saos2 cells reduces the ability of E2F-1 to bind to its consensus DNA sequence, without altering E2F-1 protein levels. These data indicate that the MDMX protein is capable of associating with E2F-1 and negatively regulating its DNA binding ability. PMID- 12532332 TI - Differential regulation of Notch signal transduction in leukaemia and lymphoma cells in culture. AB - The transduction of Notch signal plays an intricate role in cell differentiation and pathogenesis of haematological malignancies as well as in certain congenital conditions. We found no genomic changes in either gene in 34 leukaemic samples and 25 leukaemia and lymphoma cell lines. The functionality of Notch signalling was tested using HES1 gene activation. We show that Notch signalling is differentially regulated in T-acute lymphoblastic leukaemia (ALL) and B-lymphoma cells. The Notch pathway is intact in a majority of B-lymphoma cell lines, but EBNA2, which mimics notch function, can occasionally activate the pathway. In contrast, the Notch pathway is constitutively active in T-ALL. This is the first demonstration of a distinction between B-lymphomas and T-cell leukaemias in the functioning of the Notch-signalling pathway. This might be related to their pathogenesis. PMID- 12532333 TI - Sodium selenite, dietary micronutrient, prevents the lymphocyte DNA damage induced by N-nitrosodiethylamine and phenobarbital promoted experimental hepatocarcinogenesis. AB - Selenium (Se), a micronutrient, has a long history in chemoprevention of mammary and colon cancers in rodent models. Se is a current clinical trial, having shown promise in prevention of prostate and other human cancers. The mechanisms involved in the in vivo anti-carcinogenic activity of Se remain to be elucidated. In the present study, we examined the effect of sodium selenite supplementation in lymphocytes, obtained from hepatoma bearing rats on DNA damage in correlation with oxidative stress. In addition, this study examined the supplementation of Se at 4-ppm levels in the form of sodium selenite either before initiation or during initiation and/or promotion phase's increases lymphocyte Se concentrations. This in turn improves lymphocyte resistance to oxidative stress and protection against the lymphocytes DNA damage. Supplementation of Se increased lymphocyte Se concentration and reduced lymphocytes DNA damage as determined by single cell gel electrophoresis. The enzymatic antioxidants such as superoxide dismutase, glutathione peroxidase, and catalase were found to be decreased while the thiobarbituric acid reactive substances level was increased in the lymphocytes of hepatoma bearing rats. Furthermore, the reactive oxygen species such as superoxide radicals and hydroxyl radicals were also found to be high in lymphocytes. Our present results explain the understanding of unique association between anti-peroxidative effect of Se and ultimately the capability of Se to prevent cancer. PMID- 12532334 TI - Phosphorylation of proteins in neuron terminals: specificity depends on coincidental signaling. AB - We investigate the role of neuronal coincidental signaling mediated by the second messengers, on phosphorylation of three major proteins of neurosecretory vesicles. Our data show that different combinations of coincidental signaling generate specific pattern of phosphoproteins and not strictly additional effects. This suggests that an added phosphate on a site might 'mask' or 'unmask' the next sites for specific kinases and phosphatases action by inducing conformation change or protein association. We show that a function of vesicles such as the uptake of glutamate is highly regulated by coincidental signaling. PMID- 12532335 TI - Nuclear matrix localization of high mobility group protein I(Y) in a transgenic mouse model for prostate cancer. AB - Nuclear shape and the underlying nuclear structure, the nuclear matrix in cancer cells. Since the NM composition is considered to maintain nuclear shape and architecture, nuclear matrix proteins (NMPs) may be involved in transformation. Our laboratory has recently characterized a subset of NMPs that are associated with prostate cancer development in the transgenic adenocarcinoma of mouse prostate (TRAMP) model. One of the identified NMPs, E3E, has a similar molecular weight (22 kDa) with a protein known as HMGI(Y). HMGI(Y) belongs to a group of non-histone and chromatin-associated proteins, high-mobility-group (HMG) proteins, and it has been shown to associate with the NM. HMGI(Y) has been reported to be elevated in different types of cancer including prostate cancer. In this study, we examined the expression of HMGI(Y) protein in the NMP composition of the TRAMP model during the progression from normal to neoplasia. The expression of HMGI(Y) in the NMP extracts of three prostatic epithelial cell lines derived from a 32-week TRAMP mouse: TRAMP-C1, TRAMP-C2, and TRAMP-C3 was also examined. Using both one-dimensional and high-resolution two-dimensional immunoblot analyses, we found that: (i) HMGI(Y) is a nuclear matrix protein expressed as two protein bands with MW of 22-24 kDa and (ii) HMGI(Y) expression is correlated with neoplastic and malignant properties in late stage TRAMP prostate tumors. Overall, these findings support the evidence that HMGI(Y) can be utilized as a marker and prognostic tool for prostate cancer. PMID- 12532336 TI - Effects of 1,25-dihydroxyvitamin D3 on the distribution of androgen and vitamin D receptors in human prostate neonatal epithelial cells. AB - Although many studies have examined the mechanisms of 1,25-dihydroxyvitamin D(3) (calcitriol or 1,25 D) action in different prostate cancer cell lines, little is known regarding the influence of this steroid on the normal prostate. The presence of both VDR and AR in normal prostatic tissues raises the distinct possibility of an important role for this hormone in the normal gland. In order to ascertain the possible role of 1,25 D on both AR and VDR in the normal prostate, the effects of calcitriol and dihydrotestosterone (DHT) on the normal human neonatal prostatic epithelial cell line, 267B-1, were examined. These studies were approached by focusing on how 1,25 D in the presence or absence of DHT affects the distribution of AR and VDR in the cytoplasmic and nuclear compartments of the cells in terms of their protein levels and DNA binding activities. Immunoblot analyses show that 1,25 D increases the AR protein level in both the cytoplasmic and nuclear fractions but not the VDR protein level. On the other hand, the gel shift assays demonstrate that 1,25 D increases both the AR- and VDR-DNA binding activities in the nuclear fraction, whereas there is no increase in DNA binding activities in the cytoplasmic fraction. Addition of DHT along with 1,25 D does not affect the DNA binding activities of both AR and VDR. Overall, these studies suggest that 1,25 D actions on the normal prostate cells may be mediated independently through AR and VDR, respectively. PMID- 12532337 TI - Mutation analysis of the non-muscle myosin light chain kinase (MLCK) deletion constructs on CV1 fibroblast contractile activity and proliferation. AB - Smooth muscle myosin light chain kinase (MLCK) is a multifunctional molecule composed of an N-terminal actin binding domain, a central kinase domain, and C terminal calmodulin- and myosin-binding domains. We previously cloned and characterized a novel MLCK isoform from endothelial cells (EC MLCK) consisting of 1,914 amino acids displaying a higher molecular weight (210 kDa) and a novel amino-terminal stretch of 922 amino acids not shared by the smooth muscle isoform (smMLCK, 150 kDa). To further define the role of specific EC MLCK motifs in endothelial and non-muscle cells, we constructed two epitope-tagged EC MLCK deletion mutants in mammalian expression vectors lacking either the C-terminal auto-inhibitory and calmodulin-binding domain (EC MLCK1745) or the ATP-binding site (EC MLCKATPdel). Expression of EC MLCK1745 in CV1 fibroblasts showed increased basal actin stress fiber formation, which was markedly enhanced after tumor necrosis factor (TNF-alpha) or thrombin treatment. Distribution of EC MLCK1745 was largely confined to stress fibers, cortical actin filaments, and focal adhesion contacts, and co-localized with myosin light chains (MLCs) diphosphorylated on Ser(19) and Thr(18). In contrast, immunofluorescence staining demonstrated that EC MLCKATPdel abolished thrombin- and TNFalpha-induced stress fiber formation and MLC phosphorylation, suggesting this kinase-dead mutant functions as a dominant-negative MLCK construct, thereby confirming the role of EC MLCK in stress fiber formation. Finally, we compared the serum-stimulated growth rate of mutant MLCK-transfected fibroblasts to sham controls, and found EC MLCK1745 to augment thymidine incorporation whereas EC MLCKATPdel reduced CV1 growth rates. These data demonstrate the necessary role for MLCK in driving the contractile apparatus via MLC phosphorylation, which can alter fibroblast growth and contractility. PMID- 12532341 TI - Protein design at the crossroads of biotechnology, chemistry, theory, and evolution. PMID- 12532338 TI - Immunohistochemical analysis of Bin1/Amphiphysin II in human tissues: diverse sites of nuclear expression and losses in prostate cancer. AB - The Bin1/Amphiphysin II gene encodes at least seven alternately spliced adapter proteins that have been implicated in membrane dynamics and nuclear processes. Nuclear localized Bin1 polypeptides have tumor suppressor and proapoptotic activities, suggesting that Bin1 may suppress cancer in tissues where nuclear expression may occur. One question is the extent to which human tissues express nuclear Bin1 isoforms. A secondary issue has been the need for a specific antibody that can detect all the splice isoforms expressed by the human, mouse, and rat Bin1 genes. Using a novel mouse monoclonal antibody with these characteristics, we performed an immunohistochemical analysis of Bin1 expression in a panel of normal human tissues. We also compared the expression profile of Bin1 in normal or malignant tissues derived from human prostate, where Bin1 is a candidate tumor suppressor gene. In brain, a distinct nuclear staining pattern overlapped with a cytosolic staining pattern present in certain layers of the cerebral cortex and cerebellum. Bone marrow cells displayed mainly nuclear localization whereas peripheral lymphoid cells exhibited mainly cytosolic localization. In several epithelial tissues, nuclear or nucleocytosolic staining patterns were displayed by basal cells in skin, breast, or prostate, whereas cytosolic or plasma membrane-associated staining patterns were noted in gastrointestinal cells. Interestingly, a striking gradient of expression was observed in gastrointestinal epithelia, particularly in the large intestine, with the strongest staining displayed by cells destined to undergo apoptosis at the villus tip. In prostate, Bin1 staining was frequently absent in cases of primary prostate adenocarcinoma. This study used a novel reagent to document the extent of expression of nuclear Bin1 isoforms, which exhibit cancer suppression and proapoptotic activity in human cells. PMID- 12532342 TI - Recent advances in applications of room-temperature ionic liquid/supercritical CO2 systems. PMID- 12532343 TI - Asymmetric catalytic aza-Henry reactions leading to 1,2-diamines and 1,2 diaminocarboxylic acids. PMID- 12532345 TI - Tuning the regioselectivity in the palladium(II)-catalyzed isomerization of alkylidene cyclopropyl ketones: a dramatic salt effect. PMID- 12532344 TI - Homogeneous and heterogeneous catalysis: bridging the gap through surface organometallic chemistry. AB - Surface organometallic chemistry is an area of heterogeneous catalysis which has recently emerged as a result of a comparative analysis of homogeneous and heterogeneous catalysis. The chemical industry has often favored heterogeneous catalysis, but the development of better catalysts has been hindered by the presence of numerous kinds of active sites and also by the low concentration of active sites. These factors have precluded a rational improvement of these systems, hence the empirical nature of heterogeneous catalysis. Catalysis is primarily a molecular phenomenon, and it must involve well-defined surface organometallic intermediates and/or transition states. Thus, one must be able to construct a well-defined active site, test its catalytic performance, and assess a structure-activity relationship, which will be used, in turn-as in homogeneous catalysis-to design better catalysts. By the transfer of the concepts and tools of molecular organometallic chemistry to surfaces, surface organometallic chemistry can generate well-defined surface species by understanding the reaction of organometallic complexes with the support, which can be considered as a rigid ligand. This new approach to heterogeneous catalysis can bring molecular insight to the design of new catalysts and even allow the discovery of new reactions (Ziegler-Natta depolymerization and alkane metathesis). After more than a century of existence, heterogeneous catalysis can still be improved and will play a crucial role in solving current problems. It offers an answer to economical and environmental problems faced by industry in the production of molecules (agrochemicals, petrochemicals, pharmaceuticals, polymers, basic chemicals). PMID- 12532346 TI - Mono-tetrathiafulvalene calix[4]pyrrole in the electrochemical sensing of anions. PMID- 12532347 TI - Towards multistep nanostructure synthesis: programmed enzymatic self-assembly of DNA/gold systems. PMID- 12532348 TI - Catalytic oxidation of alcohols in water under atmospheric oxygen by use of an amphiphilic resin-dispersion of a nanopalladium catalyst. PMID- 12532349 TI - Crystalline structures of Sb4 molecules in antimony thin films. PMID- 12532350 TI - Analogues of neuropeptide Y containing beta-aminocyclopropane carboxylic acids are the shortest linear peptides that are selective for the Y1 receptor. PMID- 12532351 TI - [(Eta5-C5H5)(OC)3V=B=N(SiMe3)2: a half-sandwich complex with a terminal borylene ligand. PMID- 12532352 TI - Archaea analogue thiolipids for tethered bilayer lipid membranes on ultrasmooth gold surfaces. PMID- 12532353 TI - Multiple ion cores in anthracene anion clusters. PMID- 12532354 TI - Polyhydrido(silylene)osmium and silyl(dinitrogen)ruthenium products through redistribution of phenylsilane with osmium and ruthenium pincer complexes. PMID- 12532355 TI - Soil fungal hyphae bind and attack asbestos fibers. PMID- 12532356 TI - High-spin wheel of a heptanuclear mixed-valent Fe(II,III) complex. PMID- 12532357 TI - Beyond the icosahedron: the first 13-vertex carborane. PMID- 12532358 TI - Macrocycle synthesis by olefin metathesis on a nanosized, shape-persistent tricationic platinum template. PMID- 12532359 TI - [3+2] and [4+1] cycloaddition reactions of Fischer alkoxy(alkenyl)carbene complexes with electronically neutral 1,3-dienes. PMID- 12532360 TI - A catalytic and enantioselective desymmetrization of meso cyclic allylic bisdiethylphosphates with organozinc reagents. PMID- 12532362 TI - New year reflections: the integration of practice and research. PMID- 12532363 TI - Teen pregnancy, motherhood, and unprotected sexual activity. AB - The sexual behaviors and attitudes toward condom use of adolescent mothers (N = 572) from ethnic minority groups were examined. Constructs from social cognitive theory (SCT), the theory of reasoned action (TRA), and the theory of planned behavior (TPB; e.g., intentions to use condoms, self-efficacy, outcome expectancies) were measured with questionnaires. Measures of AIDS and condom-use knowledge and selected psychosocial, behavioral, and demographic variables were included. Many adolescents reported early onset of sexual activity, multiple lifetime sexual partners, substance use, and childhood sexual or physical abuse. Only 18% stated a condom was used at last intercourse. Using hierarchical regression analysis, 13% of the variance for factors associated with unprotected sex was accounted for by TRA constructs. Other variables contributed an additional 17% of the variance. Unprotected sex was associated with behavioral intentions to use condoms, pregnancy, having a steady partner, more frequent church service attendance, and ever having anal sex. Findings support the urgent need for broad-based HIV prevention efforts for adolescent mothers that build on theoretical concepts and address the realities of their lives. PMID- 12532364 TI - Testing the relationship between self-agency and enactment of health behaviors. AB - A theoretical review provides a rationale for examining self-agency as a developmental foundation underlying processes of self-regulated change and a potential moderator of intervention effectiveness among participants in a nurse home-visitation program. Self-agency is defined as the conceptual understanding of self as an agent capable of shaping motives, behavior, and future possibilities (Damon & Hart, 1991). Availability of a sample of 186 mothers who received nurse home visitation provided an opportunity to test the relationship between participant self-agency and enactment of targeted health behaviors. Self agency items from the Pearlin Mastery Scale (1978) were used to differentiate mothers who endorsed self-agency from those who did not. Consistent with the theoretical premise, mothers who endorsed self-agency at an established threshold were significantly more likely to enact health behaviors promoted during nurse visitation. Results provide support for the relationship between the development of self-agency and enactment of health behaviors targeted by a nurse home visitation program. PMID- 12532365 TI - Adaptation of the postpartum support questionnaire for mothers with multiple sclerosis. AB - The purposes of this study were to determine the dimensionality of the Postpartum Support Questionnaire adapted for mothers with multiple sclerosis through factor analytic techniques and to provide construct validity for the factored dimensions. One hundred and seventy-four mothers with multiple sclerosis comprised the sample. Using a priori criteria, a three-factor structure, resulting from principal components analysis with an orthogonal rotation consisting of 24 of the 28 items, was found to best represent the dimensionality of the instrument. The three support factors-emotional, instrumental, and informational-showed acceptable coefficient alpha reliabilities at the 1-, 3-, and 6-month administrations. Evidence of construct validity was shown in that, as hypothesized, (a) mothers with comparatively lower emotional distress reported higher levels of emotional and instrumental support, and (b) mothers with one child reported receiving more informational support during the 6-month postpartum period than did mothers with more than one child. The results support the use of the 24-item Postpartum Support Questionnaire in assessing the adequacy of these specific dimensions of social support needed by mothers with multiple sclerosis during the postpartum period. PMID- 12532366 TI - Effects of Tellington touch in healthy adults awaiting venipuncture. AB - Many natural-healing modalities administered by professional nurses are provided without adequate scientific scrutiny. Tellington touch (TTouch), a form of gentle physical touch originally developed for the calming of horses, is an emerging nursing intervention. However, the safety and efficacy of human TTouch has not yet been established. The purpose of this study, which used a pretest, posttest repeated-measures control group design, was to identify patterns of mean blood pressure (MBP), heart rate (HR), state anxiety (SA), and procedural pain (PP) in healthy adults receiving a 5-min intervention of TTouch (n = 47) just before venipuncture versus a no-touch control group (n = 46). There were statistically and clinically significant decreases in the TTouch group in MBP and HR. There were no significant differences between groups in SA and PP. Further research is essential to determine the safety and efficacy of this modality for acutely or critically ill patients. PMID- 12532367 TI - Effectiveness of a pilot project of discharge planning in Taiwan. AB - The purpose of this study was to examine the effects of a nurse-designed discharge-planning project in a teaching hospital in Taiwan. A before-and-after quasi-experimental design was used. Craniotomy (n = 112) and stroke (n = 171) patients were recruited and divided into control and intervention groups. Outcome indicators included length of hospital stay, change in activities of daily living (ADL), rate of nursing home placement, rate of unplanned readmission, and level of satisfaction. In the stroke sample length of hospital stay was shorter in the intervention group than in the control group. In the craniotomy sample unplanned readmission was less frequent in the intervention group than in the control group. No differences were found between the control and intervention groups in other outcome indicators. PMID- 12532368 TI - Methods to document semantic equivalence of a translated scale. AB - To ensure the semantic equivalence of the Postpartum Depression Screening Scale (PDSS)-Spanish Version, multiple methods were used, including back-translation, the committee approach, pretest techniques, and alternate forms equivalence. The back-translation process involved eight translators who represented the four predominant Hispanic groups in the United States. In the committee approach the Hispanic translators convened together to discuss the results of the back translations and to come to group consensus on problematic items. In pretesting the PDSS-Spanish Version was field-tested with five Hispanic mothers, and it was determined that it accurately captured the connotative meanings of the screening scale. Alternate forms equivalence of the two language versions of the PDSS was assessed with a sample of 30 bilingual Hispanic mothers. Alpha reliability estimates ranged from 0.94 to 0.99. Though lengthy, this systematic procedure was needed to guarantee the production of a fully equivalent linguistic version of the PDSS. PMID- 12532369 TI - Ethological methods to develop nursing knowledge. AB - Researchers from various fields use ethological methods to systematically observe, describe, and measure animal and human nonverbal behavior. The purpose of this article is to argue that their application in nursing will benefit development of descriptive-level knowledge about complex behavioral phenomena. To advance the argument for applying these methods in nursing, we examine the compatibility of the philosophical assumptions underlying ethology with nursing, assess if ethology can help nursing achieve some of its aims, and determine the benefits of using ethology when observation of a phenomenon is required. Neonatal pain is used to illustrate how ethology can be used to develop descriptive-level nursing knowledge and midrange theory. PMID- 12532370 TI - Barriers to nonviral gene delivery. AB - The use of various synthetic lipids and polymers to deliver DNA for gene therapy applications has been the subject of intense examination for the last 15 years. Our understanding of the processes involved in the delivery of DNA, although still limited, can be described in terms of specific physical and chemical barriers encountered along the delivery pathway. Successful engagement of this pathway involves avoiding inactivation in the extracellular compartment and initial favorable interactions with the cell surface. Internalization of the delivery system by endocytosis results in a poorly defined endosomal trafficking process which, if not escaped, leads to degradation of the therapeutic DNA in lysosomes. For the small fraction of material that is able to escape this vesicular trafficking pathway, the cytosol provides additional physical and metabolic barriers to further trafficking to the nucleus. Finally, nuclear uptake has been demonstrated to be a significant barrier to gene delivery. In this review, we outline in greater detail the various processes involved in each step and describe various formulation variables that have been explored to overcome these delivery barriers to nonviral gene delivery. PMID- 12532371 TI - Thermal stability of vaccines. AB - Worldwide vaccination programs against infectious diseases and toxins are estimated to save approximately 3 million lives yearly. Tragically, however, another 3 million individuals (primarily children) die of vaccine-preventable diseases. A significant portion of this problem results from the thermal instability of many of the currently used vaccines. This review argues that modern methods of physical and chemical analysis permit for the first time characterization of the degradative pathways of thermally labile vaccines. A rigorous description of these pathways permit a more rational and systematic approach to the stabilization of vaccines. A direct result of the replacement of currently employed, primarily empirical, approaches to vaccine stabilization with a more molecular-based methodology should be the development of more universally available vaccinations against life-threatening diseases. This has the potential to have a dramatic impact on world health. PMID- 12532372 TI - A kinetic analysis of substance P trafficking. AB - The potential for administering substance P (SP) nocitoxins for the treatment of chronic pain has been identified. To characterize treatment protocols for the spinal cord or elsewhere, binding/internalization of these compounds at the cellular targets must be understood quantitatively. Thus, a kinetic model of SP binding and intracellular trafficking has been developed from data. The eight differential equation model describes surface binding between SP and neurokinin 1 receptor, clathrin-mediated endocytosis followed by spatial translation to a perinuclear endosome where SP is sorted from its receptor, SP degradation in late endosomes/early lysosomes, and return of sorted receptor to plasma membrane via recycling endosomes. With suitably optimized parameters, the model accounts for the kinetics of total, membrane-associated, and internalized SP in cells continuously exposed to SP, as well as the fractions of internalized SP remaining intact at 30 and 60 min. Simultaneously, the model accounts for the kinetics of internalization and receptor recycling after SP preloading of membrane and subsequent exposure to SP-free media. Rate constants (min(-1)) are: 0.034 +/- 0.004 (receptor off-rate), 0.15 +/- 0.03 (internalization), 0.048 +/- 0.003 (exit from sorting endosome), 0.062 +/- 0.008 (exit of labeled SP amino acids from prelysosome), and 0.029 +/- 0.004 (receptor return from recycling endosome to plasma membrane). The SP kinetics resemble those of transferrin and its receptor at the internalization step, but are several-fold slower in the sorting and recycling steps. PMID- 12532373 TI - Subtleties in crystal structure solution from powder diffraction data using simulated annealing: ranitidine hydrochloride. AB - Recent advances in crystallographic computing and availability of high-resolution diffraction data have made it relatively easy to solve crystal structures from powders that would have traditionally required single crystal samples. The success of direct space methods depends heavily on starting with an accurate molecular model. In this paper we address the applicability of using these methods in finding subtleties such as disorder in the molecular conformation that might not be known a priori. We use ranitidine HCl as our test sample as it is known to have a conformational disorder from single crystal structural work. We redetermine the structure from powder data using simulated annealing and show that the conformational disorder is clearly revealed by this method. PMID- 12532374 TI - Effect of flavonoids on MRP1-mediated transport in Panc-1 cells. AB - The purpose of this study was to identify the effects of dietary flavonoids, which are present in fruits, vegetables, and plant-derived beverages, on the transport of daunomycin (DNM) and vinblastine (VBL) in Panc-1 cells. Panc-1 is a human pancreatic adenocarcinoma cell line, which expresses Multidrug Resistance Associated Protein1 (MRP1). The 2-h accumulation of (3)H-DNM and (3)H-VBL was determined in the presence and absence of 22 flavonoids. Biochanin-A, genistein, quercetin, chalcone, silymarin, phloretin, morin, and kaempferol, at 100 microM concentrations, all significantly increased the accumulation of both DNM and VBL in Panc-1 cells, with morin increasing DNM and VBL accumulation by 546 +/- 50% (mean +/- SE, n = 9) and 553 +/- 37% (n = 9), respectively. Fisetin treatment significantly decreased the accumulation of both DNM and VBL. Concentration dependent studies demonstrated significant effects on VBL accumulation at 50 microM, but not at 10 microM concentrations, except for chalcone that was effective at a 10 microM concentration. Following a 24-h incubation, there were no changes in MRP1 membrane expression or glutathione-S-transferase activity in cells. Cellular glutathione (GSH) concentrations were significantly decreased following a 2-h incubation with biochanin A, chalcone, genistein, phloretin, quercetin, and silymarin, and following a 24-h incubation with biochanin A, chalcone, genistein, and phloretin. These results therefore indicate that the flavonoids morin, chalcone, silymarin, phloretin, genistein, quercetin, biochanin A, and kaempferol can inhibit MRP1-mediated drug transport, effects that may involve binding interactions with MRP1, as well as modulation of GSH concentrations. PMID- 12532375 TI - Estradiol-progesterone interaction during the preparation of vaginal rings. AB - An unexpected enhanced release, in vitro, of estradiol (E2) was observed on the preparation of vaginal rings containing E2 and progesterone (P) in a silicone elastomer. The present work deals with exploring the reason(s) behind this enhanced E2 release. The effect of the ring design (i.e., putting P and E2 in the same compartment or in adjacent or separate compartments) was studied. The effects of the curing temperature as well as the curing time were also investigated. The possible interaction(s) between P and E2 on simple heating of their mixtures was investigated using infrared (IR), differential scanning calorimetry (DSC), and nuclear magnetic resonance (NMR) techniques. Also, the dissolution behavior of P, E2, and their mixture before and after heating was studied. The ring design, with respect to the position of the steroid layer(s), affected the release of P and E2 from the vaginal rings. Curing the rings at higher temperatures (>/=140 degrees C) for >/=30 min resulted in an enhanced release of the steroids, especially E2. The IR, DSC, phase diagram, and NMR results indicate that an interaction between P and E2, leading to the formation of a molecular complex, took place. It was concluded that putting P and E2 in the same compartment and curing by heating at a high temperature and for an extended time promoted this kind of interaction. The greater hydrophobicity of the interaction product, relative to that of E2, was considered the main reason behind the enhanced in vitro release of E2 from the vaginal rings. PMID- 12532376 TI - Effects of types of sugar on the stabilization of protein in the dried state. AB - The effects of various sugars on the structural stabilization of protein during freeze-drying were investigated. The degree of native structure of protein that was freeze-dried and rehumidified at constant relative humidities (RHs) was evaluated by measurement of the alpha-helix content by Fourier-transform infrared spectroscopy. Bovine serum albumin (BSA) and several types of sugars, including sucrose, trehalose, and dextrans, were used as a model protein and sugars, respectively. The glass transition temperature, T(g), for the amorphous sugar samples was measured by differential scanning calorimetry (DSC) to characterize the structural stability of sugars. The dependence of the alpha-helix content (C(alpha-helix)) of BSA on the sugar content (c(sugar)) could, in most cases, be represented by a Langmuir-type equation: C(alpha-helix) = K x (C(alpha-helix) (max) - C(alpha-helix) (0)) x c(sugar)/(1 + K x c(sugar)) + C(alpha-helix) (0), where K is a constant, indicating the ability of amorphous sugar matrix to embed protein, and C(alpha-helix) (0) and C(alpha-helix) (max) indicate the alpha-helix content in the absence of sugar and saturating levels of sugar, respectively. The preservation effects of the sugars could be characterized by K and C(alpha-helix) (max). Both K and C(alpha-helix) (max) values tended to be higher with decreasing T(g) values for the amorphous sugar, probably because an amorphous sugar matrix with lower T(g) values is structurally more flexible. The rehumidification of protein that was freeze-dried in the presence of sugar induced the refolding of protein structure, whereas the protein dried alone did not show any recovery of its native structure. PMID- 12532377 TI - Two reaction sites of a spin label, TEMPOL (4-hydroxy-2,2,6,6 tetramethylpiperidine-N-oxyl), with hydroxyl radical. AB - The exposure of 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPOL) to hydroxyl radical ((.)OH) led to the appearance of a new triplet ESR signal along with a reduction in the intensity of the TEMPOL signal. The observed spectrum was in good agreement with the spectrum simulated with 4-oxo-2,2,6,6 tetramethylpiperidine-N-oxyl (TEMPON) and TEMPOL, and the addition of sodium borohydride to the product solution led to the disappearance of the new signal. These results indicate the formation of TEMPON during the reaction. A comparison of the concentration of TEMPON formed with that of nitroxyl compounds that lost the electron spin resonance (ESR) signal showed that the rate of TEMPON formation is less than one-tenth that of the paramagnetism loss caused by (.)OH in the pH range 6.5-8.5. These results indicate that TEMPOL has two reaction sites with (.)OH; the nitroxyl group and the 4-position of the piperidine ring. The reaction at the 4-position should contribute to the (.)OH scavenging activity of TEMPOL together with that at the nitroxyl group, although the rate of the former reaction is relatively low. PMID- 12532378 TI - Influence of phytostanol phosphoryl ascorbate (FM-VP4) on insulin resistance, hyperglycemia, plasma lipid levels, and gastrointestinal absorption of exogenous cholesterol in Zucker (fa/fa) fatty and lean rats. AB - The purpose of this investigation was to determine the effects of Phytostanol Phosphoryl Ascorbate (FM-VP4) on insulin resistance, hyperglycemia, plasma lipid levels, body weight, and gastrointestinal absorption of exogenous cholesterol in Zucker (fa/fa) fatty and lean rats. A group of 12 age-matched male obese (n = 6) and lean (n = 6) Zucker rats were administered 250 mg/kg twice a day (as 2% FM VP4 in drinking water) for 30 consecutive days. Fasted blood samples prior to and following treatment were taken from all rats for glucose, lipid, insulin, and leptin determination. An oral glucose tolerance test was also carried out at the end of the treatment protocol. In addition, male obese (n = 7) and lean (n = 8) Zucker rats were coadministered a single oral gavage of [(3)H]cholesterol plus cold cholesterol with or without FM-VP4 (20 mg/kg) dissolved in Intralipid and the plasma concentration of the radiolabel was determined 10 h following the dose. FM-VP4 30-day treatment did not alter body weight, morning glucose, insulin, lipids, and leptin concentrations. There was no alteration in glucose tolerance in the nondiabetic, normoglycemic lean group; however, there was a highly significant improvement in glucose tolerance in the fatty group following FM-VP4 treatment. In addition, the insulin response to oral glucose showed no significant change in nondiabetic lean rats, whereas there was a change in the insulin secretory profile in the fatty group following FM-VP4 treatment. Furthermore, following a single oral gavage of FM-VP4 resulted in a significant decrease in the percentage of radiolabeled cholesterol absorbed. These findings suggest that FM-VP4 treatment to fatty Zucker rats could result in increased glucose responsiveness of the insulin secreting pancreatic beta cells. Furthermore, our findings suggest that FM-VP4 may only be effective presystemically. Systemic administration of FM-VP4 is warranted to determine the therapeutic potential of this effect. PMID- 12532379 TI - Noninvasive monitoring of local drug release using X-ray computed tomography: optimization and in vitro/in vivo validation. AB - In vivo release profiles of drug-loaded biodegradable implants were noninvasively monitored and characterized using X-ray computed tomography (CT). The imaging method was adapted and optimized to quantitatively examine the release of an active agent from a model cylindrical PLGA device (the millirod) into rabbit livers over 48 h. Iohexol, a CT contrast agent, served as a model drug; optimization of CT acquisition parameters yielded a sensitivity of 0.21 mg/mL (or 95 microg iodine/mL) for this agent. In vitro validation of the method was carried out by tracking release of iohexol in gelatin gel phantoms. In vivo release in rabbit livers was characterized through quantitative analysis of CT images and compared with UV-Vis analysis of the explanted devices at three implantation times. After correction for respiratory motion, CT analysis correlates well with the extracted iohexol data at all time points. The percent error between the actual and experimental image data was below 10%. This study demonstrates the potential of using computed tomography to noninvasively quantify the rate of agent release from controlled delivery devices in vivo. PMID- 12532380 TI - Mechanistic study of alkyl azacycloheptanones as skin permeation enhancers by permeation and partition experiments with hairless mouse skin. AB - In previous studies (Yoneto et al., 1995. J Pharm Sci 84:312-317; Kim et al., 1992. Int J Pharm 80:17-31; and Warner et al., 2001. J Pharm Sci 90:1143-53), the transport enhancing effects of four homologous series of enhancers-the n alkanols, 1-alkyl-2-pyrrolidones, 1,2-alkanediols, and N,N-dimethylalkanamides - on the transport of steroidal permeants across hairless mouse skin (HMS) were investigated. Isoenhancement concentrations are defined as the aqueous concentrations for which different enhancers induce the same extent of permeant transport enhancement, E, for the lipoidal pathway of the stratum corneum (SC). Our studies have shown that the E = 10 isoenhancement concentrations of these four homologous series were nearly the same when compared at the same n-alkyl group chain length and therefore that the contribution of the polar head group toward the enhancer potency was found to be essentially constant. In the present study, we have determined the isoenhancement concentrations (E = 10) for the 1 alkyl-2-azacycloheptanone series [1-butyl-2-azacycloheptanone (BAZ), 1-hexyl-2 azacycloheptanone (HAZ), and 1-octyl-2-azacycloheptanone (OAZ)] and compared the results with those of the previously studied four homologous series. We have found that the E = 10 isoenhancement concentrations (aqueous phase concentrations) of the 1-alkyl-2-azacycloheptanones (Azs) are around 10 times lower than those for the previously studied four homologous series when compared at the same alkyl group chain length. This indicates an approximately 10 times higher potency of Azs. This finding was a point of interest because the polar group of Azs is similar to that of 1-alkyl-2-pyrrolidones (Aps). To further probe the nature of the mechanism of action of the Azs and Aps and to better understand the lower E = 10 isoenhancement concentrations found with the Azs, it was decided (a) to determine the equilibrium partitioning (uptake) of the Azs and the Aps from the aqueous phase into the HMS SC at E = 10, and (b) to determine the equilibrium partitioning (uptake) of a surrogate permeant, estradiol (E2beta), into the SC in the absence of and in the presence of Azs and Aps at E = 10. The following were the outcomes from the two partitioning studies. Firstly, at the E = 10 isoenhancement concentrations, the extent of partitioning (uptake) of the Azs and Aps into the intercellular lipids of the HMS SC was found to be approximately the same, even though the E = 10 isoenhancement concentrations (aqueous phase concentrations) of the Aps were around 10 times greater than those of the Azs. We interpret this to mean (whereas the potencies of the Azs are around ten times greater than those of the Aps when related to their aqueous concentrations) that the potencies of the two enhancer series are about the same when expressed in terms of their concentrations in the intercellular lipid phase of the SC. Another outcome of the partitioning studies has been the finding that the extent of partitioning into the intercellular lipids of the SC at E = 10 isoenhancement conditions for both the Azs and Aps is essentially independent of the n-alkyl chain length (from butyl to octyl). A third result from these experiments has been that the partitioning of E2beta (the surrogate permeant) into the HMS SC under E = 10 isoenhancement concentration conditions is approximately the same with the Aps and Azs as enhancers. For both the Aps and Azs, the E2beta SC partitioning enhancement was found to be in the range of 5-6 at E = 10. This comparable partitioning enhancement for E2beta in the presence of Aps and Azs at E = 10 suggests that the same mechanism was involved and that these enhancers act, in part but to a significant extent, by inducing a higher partitioning tendency of the permeant into the transport rate-limiting lipoidal domains of the SC. (c) 2003 Wiley-Liss, Inc. and the American Pharmaceutical Association J Pharm Sci 92:297-310, 2003 PMID- 12532381 TI - Regional intestinal absorption of FITC-dextran 4,400 with nanoparticles based on beta-sitosterol beta-D-glucoside in rats. AB - Nanoparticles (NP) are potential carriers for drug delivery to the targeted intestine. NP based on beta-sitosterol beta-D-glucoside (Sit-G) enhanced the colon-specific absorption of FITC-dextran 4,400 (FD-4), because the concentration dependent increase of bioavailability appeared in only the colon. In a permeation study, the absorption enhancement in the colon was suppressed in the following conditions: (1) the addition of Sit-G NP to serosa; (2) a permeation study at 4 degrees C; (3) the addition of endocytosis inhibitor, cytochalasin B. NP based on sitosterol, the aglycon of Sit-G, did not increase the FD-4 colonic permeation. The addition of Sit-G NP to the mucosal side induced a decrease of transepithelial resistance (TEER), but this phenomenon was suppressed by an inhibitor of Na(+)-dependent specific glucose transporter, phrolidzin, which did not affect FD-4 permeation. These findings suggested that absorption enhancement by Sit-G NP may not be due to opening of a tight junction, but might be related to endocytosis via glucose residue of Sit-G. PMID- 12532382 TI - Stabilization of alum-adjuvanted vaccine dry powder formulations: mechanism and application. AB - Studies were performed to elucidate the mechanism of alum gel coagulation upon freezing and drying and its relationship to vaccine potency loss and to develop a novel freeze-drying process for the production of stable alum-adjuvanted vaccine formulations suitable for conventional needle injection and epidermal powder immunization (EPI). The alum hydroxide-adjuvanted hepatitis-B surface antigen (Alum-HBsAg) and the alum phosphate-adjuvanted diphtheria and tetanus toxoids (Alum-DT) were dehydrated by freeze drying (FD), spray drying (SD), air drying (AD), or spray freeze drying (SFD). After drying by FD, SD, or AD, alum gels coagulated when examined by optical microscopy and particle size analysis. In addition, desorption of antigen molecules from the coagulated when examined by optical microscopy and particle size analysis. In addition, desorption of antigen molecules from the coagulated alum gel upon reconstitution appeared to be difficult, as indicated by attenuated band intensity on SDS-PAGE. In contrast, SFD alum gels turned a homogenous suspension upon reconstitution, suggesting minimal alum coagulation. In the mouse model, the in vivo immunogenicity of SFD Alum-HBsAg was preserved, whereas the FD Alum-HBsAg suffered significant immunogenicity loss. Grinding of coagulated FD Alum-HBsAg into smaller particles could partially recover the immunogenicity. In a guinea pig study using EPI, the SD Alum-DT formulation was not immunogenic, but the SFD Alum-DT formulations had a vaccine potency comparable to that of the untreated DT administered by I.M. injection. Overall, the relationship of coagulation of alum gel upon reconstitution and the loss of vaccine potency was established in this study. Alum gels became highly coagulated after dehydration by spray drying and traditional freeze-drying processes. However, freezing rate played a critical role in preserving the adjuvant effect of alum and fast freezing decreased the tendency of alum coagulation. Spraying the alum gel into liquid nitrogen represents the fastest freezing rate achievable and resulted in no discernible alum coagulation. Therefore, SFD presents a novel and effective drying process for alum-adjuvanted vaccine formulations and is particularly valuable for dry powder applications such as EPI. PMID- 12532383 TI - Characterizing a drug's primary binding site on albumin. AB - Surface plasmon resonance-based biosensors can be used to directly measure the binding of small molecules to albumin. We studied 12 drugs with different molecular masses and affinities for albumin to illustrate the benefits of the technology. To examine both high- and low-affinity sites on the protein, each drug was assayed across a 10,000-fold concentration range. The affinity constants determined from the biosensor assay corresponded with affinities determined by other methods. We expanded the utility of the biosensor technology by developing protocols to characterize drug displacement from albumin. Finally, we also compared how a representative panel of drugs bound albumins from 14 species. The results illustrate how biosensors can provide detailed information about the identification and affinity of a drug's primary binding site on albumin. PMID- 12532384 TI - A systematic examination of the in vitro Ussing chamber and the in situ single pass perfusion model systems in rat ileum permeation of model solutes. AB - In situ and in vitro intestinal absorption in the rat ileum was systematically studied and mechanistically quantified in terms of permeability coefficients (P) of a series of [(3)H]steroids as model transcellular permeants, [(3)H]taurocholate utilizing the active membrane transport systems to define the aqueous boundary layer (ABL), and [(14)C]urea and [(14)C]mannitol as pore hindered paracellular diffusants. In situ single-pass perfusion experiments were performed in isolated ileal segments and blood samples were collected from the cannulated mesenteric vein. For the in vitro experiments, an excised, serosal and muscular layer-removed, ileal tissue was mounted in the Ussing chamber diffusion cells. In situ and in vitro P values versus logarithm of the partition coefficient in n-octanol/water (log K) of the steroids were characterized by a sigmoidal-shaped curve in which plateau values were attained for the highly lipophilic steroids with log K greater, similar 2.5. The in situ and in vitro transport barriers in series were viewed as ABL/mucosal epithelium and ABL/mucosal epithelium/submucosal tissue, respectively. Within this framework and the use of experimental strategies and theoretical reasoning, the transport barriers of the steroids were quantitatively delineated and the rate-determining barriers identified. In the plateau region, the analyses indicate that the in situ absorption of the lipophilic steroids was essentially ABL controlled, whereas the in vitro absorption was about equally controlled by diffusion across the ABL and submucosal tissue. The in situ and in vitro pore radii of the paracellular route were 7.2 and 9.2 A, respectively, and the difference was likely the result of perturbation of the tight junctions during the in vitro preparation of the ileal tissue. PMID- 12532385 TI - Recent advances in the prediction of blood-brain partitioning from molecular structure. AB - The widely distributed software tools Cerius2 and ACD/log D Suite have been used to develop a new method for the prediction of the ratio of concentrations of a drug in the brain and blood (BB,quantified as log BB) from structure. The performances of all known blood-brain partitioning prediction methods are compared to give an up-to-date account on their accuracy, limitations, and usefulness. It is demonstrated that the new log BB prediction method is superior to other methods with regard to low-to-medium throughput log BB prediction, whereas the C2-ADME log BB two-dimensional (2D) method seems to offer the best compromise between speed and accuracy for ultra-high throughput processing of large compound databases for log BB prediction. PMID- 12532386 TI - Pulmonary gene delivery by chitosan-pDNA complex powder prepared by a supercritical carbon dioxide process. AB - Chitosan-plasmid DNA (pDNA) complex powders as a pulmonary gene delivery system were prepared with a supercritical carbon dioxide (CO(2)) process and their in vivo activity was evaluated. The powders with mannitol as a carrier were prepared by dispersing aqueous solutions of a luciferase expression plasmid driven by the cytomegalovirus promoter (pCMV-Luc) with or without chitosan as a cationic vector in a supercritical CO(2)/ethanol admixture. The supercritical CO(2) process with a V-shaped nozzle successfully produced chitosan-pDNA powders. The addition of chitosan suppressed the degradation of pCMV-Luc during the supercritical CO(2) process and increased the yield of powders. The luciferase activity in mouse lung was evaluated after pulmonary administration of the powders or pCMV-Luc solutions. The chitosan-pDNA powders increased the luciferase activity in mouse lung compared with pCMV-Luc powders without chitosan or pCMV-Luc solutions with or without chitosan. The chitosan-pDNA powder with an N/P ratio = 5 increased the luciferase activity to 2700% of that of the pCMV-Luc solution. These results suggest that gene powder with chitosan is a useful pulmonary gene delivery system. PMID- 12532387 TI - Description of transdermal transport of hydrophilic solutes during low-frequency sonophoresis based on a modified porous pathway model. AB - Application of low-frequency ultrasound has been shown to increase skin permeability, thereby facilitating delivery of macromolecules (low-frequency sonophoresis). In this study, we sought to determine a theoretical description of transdermal transport of hydrophilic permeants induced by low-frequency sonophoresis. Parameters such as pore size distribution, absolute porosity, and dependence of effective tortuosity on solute characteristics were investigated. Pig skin was exposed to low-frequency ultrasound at 58 kHz to achieve different skin resistivities. Transdermal delivery of four permeants [mannitol, luteinizing hormone releasing hormone (LHRH), inulin, dextran] in the presence and absence of ultrasound was measured. The porous pathway model was modified to incorporate the permeant characteristics into the model and to achieve a detailed understanding of the pathways responsible for hydrophilic permeant delivery. The slopes of the log kp(p) versus log R graphs for individual solutes changed with solute molecular area, suggesting that the permeability-resistivity correlation for each permeant is related to its size. The tortuosity that a permeant experiences within the skin also depends on its size, where larger molecules experience a less tortuous path. With the modified porous pathway model, the effective tortuosities and skin porosity were calculated independently. The results of this study show that low-frequency sonophoresis creates pathways for permeant delivery with a wide range of pore sizes. The optimum pore size utilized by solutes is related to their molecular radii. PMID- 12532388 TI - Microdialysis of triamcinolone acetonide in rat muscle. AB - The objective of this study was to compare plasma and muscle concentrations of triamcinolone acetonide (TA) in the rat by microdialysis. Microdialysis experiments were carried out at steady state in rats after an initial I.V. bolus 50 mg/kg of the phosphate ester of TA (TAP) followed by 23 mg/kg/h infusion. In vivo recovery was calculated by retrodialysis. The concentration determined at steady state in microdialysate, corrected for recovery, was 2.73 +/- 0.42 microg/mL compared to 21.9 +/- 2.3 microg/mL in plasma. The pharmacokinetics of TA in plasma was described by an open two-compartment model with a terminal half life of 2.7 h. The clearance of TA in rats determined by compartmental analysis was 0.94 L/h/kg. The measured microdialysate levels of TA in muscle, corrected for recovery, were comparable to the predicted free drug levels in the peripheral compartment. Protein binding in rat plasma, measured by ultrafiltration, was 90.1%. The microdialysis in vivo recovery in muscle was similar to the in vitro recovery under stirred conditions. The results show the applicability of microdialysis to measure free tissue concentrations of TA in rats. PMID- 12532389 TI - In vitro-in vivo correlation in man of a topically applied local anesthetic agent using numerical convolution and deconvolution. AB - The aim of this study was to evaluate the relevance of the in vitro permeation method used at our laboratory in predicting in vivo dermal and transdermal absorption. Two different emulsions, a submicron oil-in-water (o/w) emulsion and a semisolid water-in-oil (w/o) emulsion, containing a model compound were investigated. The in vitro permeation rate of the compound from these emulsions was measured using static diffusion cells with human skin as membrane. The emulsions were allowed to remain in contact with the skin in the donor chamber for 15, 60, and 240 min. The study was monitored for 240 min and the steady state flux was calculated. The systemic concentration of the compound was measured in vivo as a function of time after dermal application to healthy volunteers with 15 and 60 min of application. A short-lasting i.v. infusion study in healthy volunteers was used to simulate the i.v. bolus dose. Numerical convolution was used to predict the in vivo plasma concentration of the compound while the in vivo absorption rate of the compound was estimated using numerical deconvolution. To establish correlation, the predicted in vivo flux was compared with the corresponding observed in vitro parameter after adjusting for the lag time. No major differences were seen in the systemic plasma levels between the two emulsions, which is in close agreement with the steady state flux measured in vitro. A linear correlation representing a point-to-point relationship was established for each of the investigated formulations and application times. The longer application time was predicted more accurately for both emulsions. PMID- 12532390 TI - Preparation of in situ-forming ophthalmic gels of ciprofloxacin hydrochloride for the treatment of bacterial conjunctivitis: in vitro and in vivo studies. AB - Sol-to-gel systems of ciprofloxacin hydrochloride were prepared utilizing the phase transition properties of hydroxy propyl methyl cellulose K 15 M grade (HPMC) and carbopol 934. The sol-to-gel systems were sterilized by gamma radiation and/or filtration. The sol-to-gel systems were evaluated for rheological characteristics, in vitro release behavior, microbial efficacy, in vivo release behavior, and efficacy against induced bacterial conjunctivitis in rabbits' eyes. Concentration in aqueous humor was determined and stability studies were carried out as per the ICH guidelines. The system passed the test for sterility. The sol-to-gel system exhibited a zero-order drug release pattern over 24 h in in vitro release studies. The drug was active against selected microorganisms in microbial efficacy studies. Better improvement in artificially induced bacterial conjunctivitis in rabbits' eyes was observed in animals treated with the sol-to-gel system compared with marketed eye drops. Drug concentration in aqueous humor was greater than the minimum inhibitory concentration (MIC 90) against selected microorganisms. The shelf-life of the product was >2 years. PMID- 12532391 TI - Zonula occludens toxin increases the permeability of molecular weight markers and chemotherapeutic agents across the bovine brain microvessel endothelial cells. AB - The purpose of this study was to examine the ability of Zonula occludens toxin (Zot) to reversibly open tight junctions in bovine brain microvessel endothelial cells (BBMECs) to enhance drug delivery via the paracellular pathway. Transport across BBMEC monolayers was examined for molecular weight markers and chemotherapeutic agents ([(14)C]sucrose, [(14)C]inulin, [(3)H]propranolol, [(3)H]doxorubicin, and [(14)C]paclitaxel) with Zot (0.0-4.0 microg/mL). TEER of monolayers was measured to assess effect and reversibility of Zot. Cell viability of BBMEC in the presence of Zot was assessed by trypan blue exclusion staining. Apparent permeability (P(app)), enhancement ratio (R), and percent increase in transport determined were statistically compared by ANOVA. A significant increase (p < 0.05) in P(app) was observed for the transport of [(14)C]sucrose, [(14)C]inulin, [(3)H]doxorubicin, and [(14)C]paclitaxel at a 4.0 microg/mL concentration of Zot. A significant concentration-dependent decrease in TEER was observed on treatment with Zot with rapid reversal to baseline after removal. Zot (4 micro/ml) was found to be nontoxic to the BBMECs after 2 hours incubation. In conclusion, Zot increased paracellular transport across the BBMEC in a reversible, concentration-dependent manner. Modulation of paracellular transport with Zot may be used to increase the brain permeability of potent central nervous system-active drugs, including anticancer agents. PMID- 12532392 TI - A mechanistic study of danazol dissolution in ionic surfactant solutions. AB - This study examined the dissolution mechanism of the neutral drug danazol into solutions of the ionic surfactant sodium dodecyl sulfate (SDS). The effect of counterion concentration on drug dissolution was also studied by controlling the solution ionic strength (IS). The laminar flow apparatus of Shah and Nelson was chosen to measure in vitro dissolution rates for its simulation of physiological hydrodynamics. A mathematical model was developed to test the proposed mechanism for dissolution. Transport of the dissolved drug away from the tablet surface is the slow step in the process. Two major physicochemical properties, drug solubility in surfactant solutions and the effective diffusion coefficients used in the model, were measured in separate experiments for use in the transport model. Pulsed field proton nuclear magnetic resonance spectroscopy ((1)H NMR) was used to measure the drug diffusion coefficient. Actual drug dissolution rates were determined by multiplying the measured effluent drug concentration in the aqueous medium by its flow rate. The assumption of a transport-controlled dissolution rate was tested by plotting the measured dissolution rates as a function of medium flow rate in a log-log plot. A slope of 1/3 is predicted by the model and slopes of 0.26 to 0.32 were found experimentally, suggesting that the transport controlled mechanism is accurate. The model-predicted dissolution rates were compared with the experimental data. For SDS solutions without IS control, the model calculated data are 20-35% lower than the experimental results, whereas with IS control, the error is only 0.4-4%. We believe that there is significant electrostatic interaction between micelles in processes with low IS or poor IS control. In that situation, the nuclear magnetic resonance (NMR) measured drug diffusivity would not be its actual value in the dissolution process. PMID- 12532393 TI - Hydrolysis of oxaliplatin-evaluation of the acid dissociation constant for the oxalato monodentate complex. AB - Alkaline hydrolysis of the platinum anticancer drug oxaliplatin gives the oxalato monodentate complex and the dihydrated oxaliplatin complex in two consecutive steps. The acid dissociation constant for the oxalato monodentate intermediate was determined by a kinetic approach. The pK(a) value was estimated as 7.23. The monodentate intermediate is assumed to rapidly react with endogenous compounds, resulting in a continuous conversion of oxaliplatin via the monodentate form. PMID- 12532395 TI - Synaptic Drosophila UNC-13 is regulated by antagonistic G-protein pathways via a proteasome-dependent degradation mechanism. AB - UNC-13 is a highly conserved plasma membrane-associated synaptic protein implicated in the regulation of neurotransmitter release through the direct modulation of the SNARE exocytosis complex. Previously, we characterized the Drosophila homologue (DUNC-13) and showed it to be essential for neurotransmitter release immediately upstream of vesicular fusion ("priming") at the neuromuscular junction (NMJ). Here, we show that the abundance of DUNC-13 in NMJ synaptic boutons is regulated downstream of GalphaS and Galphaq pathways, which have inhibitory and facilitatory roles, respectively. Both cAMP modulation and PKA function are required for DUNC-13 synaptic up-regulation, suggesting that the cAMP pathway enhances synaptic efficacy via DUNC-13. Similarly, PLC function and DAG modulation also regulate the synaptic levels of DUNC-13, through a mechanism that appears independent of PKC. Our results suggest that proteasome-mediated protein degradation is the primary mechanism regulating DUNC-13 levels at the synapse. Both PLC- and PKA-mediated pathways appear to regulate synaptic levels of DUNC-13 through controlling the rate of proteasome-dependent DUNC-13 degradation. We conclude that the functional abundance of DUNC-13 at the synapse, a key determinant of synaptic vesicle priming and neurotransmitter release probability, is primarily regulated by the rate of protein degradation, rather than translocation or transport, convergently controlled via both cAMP and DAG signal transduction pathways. PMID- 12532396 TI - Calcium and voltage dependent inactivation of sodium and calcium currents limits calcium influx in Helisoma neurons. AB - The control of free intracellular calcium concentration ([Ca2+]i) is necessary for cell survival because of the ubiquitous and essential role this second messenger plays in regulating numerous intracellular processes. Calcium regulation in neurons is especially vigorous because of the large calcium influx that occurs through voltage-gated channels during membrane depolarization. In this study we examined changes in ionic currents that can limit calcium influx into neurons during electrical activity. We found that the [Ca2+]i in electrically stimulated Helisoma B4 neurons initially increased to a peak and then relaxed to lower concentrations in tandem with a decline in the action potential peak voltage. The decline in [Ca2+]i and the peak action potential voltage in this sodium and calcium driven neuron was found to be a dual manifestation of I(Na) and I(Ca) inactivation. I(Na) and I(Ca) both displayed voltage dependent inactivation. Additionally, I(Na) and I(Ca) progressively inactivated at [Ca2+]i above 200 nM, concentrations readily attained in electrically stimulated B4 neurons. Calcium and voltage dependent I(Na) and I(Ca) inactivation were found to reduce calcium influx during continuous electrical stimulation by decreasing both the magnitude of I(Ca) that could be activated and the percent of the available I(Ca) that would be activated due to the diminished peak action potential voltage. Calculations based on data herein suggest that the voltage and calcium dependent I(Na) and I(Ca) inactivation that occurs during continuous electrical stimulation dramatically reduces calcium influx in this sodium and calcium driven neuron and thus limits the increase in [Ca2+]i. PMID- 12532397 TI - PDGF and FGF2 regulate oligodendrocyte progenitor responses to demyelination. AB - Acute demyelination of adult CNS, resulting from trauma or disease, is initially followed by remyelination. However, chronic lesions with subsequent functional impairment result from eventual failure of the remyelination process, as seen in multiple sclerosis. Studies using animal models of successful remyelination delineate a progression of events facilitating remyelination. A universal feature of this repair process is extensive proliferation of oligodendrocyte progenitor cells (OPs) in response to demyelination. To investigate signals that regulate OP proliferation in response to demyelination we used murine hepatitis virus-A59 (MHV-A59) infection of adult mice to induce focal demyelination throughout the spinal cord followed by spontaneous remyelination. We cultured glial cells directly from demyelinating and remyelinating spinal cords using conditions that maintain the dramatically enhanced OP proliferative response prior to CNS remyelination. We identify PDGF and FGF2 as significant mitogens regulating this proliferative response. Furthermore, we demonstrate endogenous PDGF and FGF2 activity in these glial cultures isolated from demyelinated CNS tissue. These findings correlate well with our previous demonstration of increased in vivo expression of PDGF and FGF2 ligand and corresponding receptors in MHV-A59 lesions. Together these studies support the potential of these pathways to function in vivo as critical factors in regulating remyelination. PMID- 12532398 TI - Protein expression levels of the Src activating protein AFAP are developmentally regulated in brain. AB - The Src family of nonreceptor tyrosine kinases plays an important role in modulating signals that affect growth cone extension, neuronal differentiation, and brain development. Recent reports indicate that the Src SH2/SH3 binding partner AFAP-110 has the capacity to modulate actin filament integrity as a cSrc activating protein and as an actin filament bundling protein. Both AFAP-110 and a brain specific isoform called AFAP-120 (collectively referred to as AFAP) exist at high levels in chick embryo brain. We sought to identify the localization of AFAP in mouse brain in order to identify its expression pattern and potential role as a cellular modulator of Src family kinase activity and actin filament integrity in the brain. In E16 mouse embryos, AFAP expression levels were very high and concentrated in the olfactory bulb, cortex, forebrain, cerebellum, and various peripheral sensory structures. In P3 mouse pups, overall expression was reduced compared to E16 embryos, and AFAP was found primarily in olfactory bulb, cortex, and cerebellum. AFAP expression levels were significantly reduced in adult mice, with high expression levels only detected in the olfactory bulb. Western blot analysis indicated that concentrated expression of AFAP correlates well with the AFAP-120 isoform, which appears to be a splice variant of AFAP-110. As the expression pattern of AFAP overlaps with the reported expression patterns of cSrc and Fyn, we hypothesize that AFAP is positioned to modulate signal transduction cascades that direct activation of these nonreceptor tyrosine kinases and concomitant cellular changes that occur in actin filaments during brain development. PMID- 12532399 TI - Rapsyn-mediated clustering of acetylcholine receptor subunits requires the major cytoplasmic loop of the receptor subunits. AB - During synaptogenesis at the neuromuscular junction, nicotinic acetylcholine receptors (AChRs) are organized into high-density postsynaptic clusters that are critical for efficient synaptic transmission. Rapsyn, an AChR associated cytoplasmic protein, is essential for the aggregation and immobilization of AChRs at the neuromuscular junction. Previous studies have shown that when expressed in nonmuscle cells, both assembled and unassembled AChR subunits are clustered by rapsyn, and the clustering of the alpha subunit is dependent on its major cytoplasmic loop. In the present study, we investigated the mechanism of rapsyn induced clustering of the AChR beta, gamma, and delta subunits by testing mutant subunits for the ability to cocluster with rapsyn in transfected QT6 cells. For each subunit, deletion of the major cytoplasmic loop, between the third and fourth transmembrane domains, dramatically reduced coclustering with rapsyn. Furthermore, each major cytoplasmic loop was sufficient to mediate clustering of an unrelated transmembrane protein. The AChR subunit mutants lacking the major cytoplasmic loops could assemble into alphadelta dimers, but these were poorly clustered by rapsyn unless at least one mutant was replaced with its wild-type counterpart. These results demonstrate that the major cytoplasmic loop of each AChR subunit is both necessary and sufficient for mediating efficient clustering by rapsyn, and that only one such domain is required for rapsyn-mediated clustering of an assembly intermediate, the alphadelta dimer. PMID- 12532400 TI - Organizational effects of testosterone, estradiol, and dihydrotestosterone on vasopressin mRNA expression in the bed nucleus of the stria terminalis. AB - In adulthood, male rats express higher levels of arginine vasopressin (AVP) mRNA in the bed nucleus of the stria terminalis (BST) than do female rats. We tested whether this sex difference is primarily due to differences in neonatal levels of testosterone. Male and female rats were gonadectomized on the day of birth and treated with testosterone propionate (TP) or vehicle on postnatal days 1, 3, and 5 (P1, P3, and P5). Three months later, all rats were implanted with testosterone filled capsules. Two weeks later, brains were processed for in situ hybridization to detect AVP mRNA. We found that neonatal TP treatment significantly increased the number of vasopressinergic cells in the BST over control injections. We then sought to determine the effects of testosterone metabolites, estradiol and dihydrotestosterone, given alone or in combination, on AVP expression in the BST. Rat pups were treated as described above, except that instead of testosterone, estradiol benzoate (EB), dihydrotestosterone propionate (DHTP), a combination of EB and DHTP (EB+DHTP), or vehicle was injected neonatally. Neonatal treatment with either EB or EB+DHTP increased the number of vasopressinergic cells in the BST over that of DHTP or oil treatment. However, treatment with DHTP also significantly increased the number of vasopressinergic cells over that of oil treatment. Hence, in addition to bolstering evidence that estradiol is the more potent metabolite of testosterone in causing sexual differentiation of the brain, these data provide the first example of a masculinizing effect of a nonaromatizable androgen on a sexually dimorphic neuropeptide system. PMID- 12532401 TI - Regulation of ganglion cell production by Notch signaling during retinal development. AB - Although progenitor cells in developing vertebrate retina are capable of producing all retinal cell types, they are competent to produce only certain cell types at a given time, and this competence changes as development progresses. We asked whether a change in progenitor cell competence is primarily responsible for ending production of a specific cell type, the retinal ganglion cell. Reducing Notch expression using an antisense oligonucleotide in vitro or in vivo increased ganglion cell genesis. The antisense treatment could reinitiate ganglion cell genesis after it had terminated in a region of the retina, but only for a brief period. The failure of the Notch antisense treatment to reinitiate ganglion cell production after this period was not due to the lack of receptor or ligand expression, as both Notch-1 and Delta-1 were still expressed. The failure of the Notch antisense treatment to reinitiate ganglion cell production is consistent with the suggestion that the intrinsic competence of progenitor cells changes as development progresses. Because reducing Notch signaling can reinitiate ganglion cell production for a brief period after ganglion cell production has normally ceased, it appears that ganglion cell production initially ends in a region of the retina because of cell-cell interactions and not because progenitor cells lose the competence to make ganglion cells. Notch signaling appears to temporarily prevent production of ganglion cells in a region, while some other signal must initiate a change in progenitor cell competence, thus permanently ending the possibility of further ganglion cell production. PMID- 12532404 TI - A unique mushroom body substructure common to basal cockroaches and to termites. AB - The mushroom bodies of the cockroach Periplaneta americana are made up of intrinsic neurons (class I and class II Kenyon cells) with dendrites in a dorsal calyx and axons that bifurcate into medial and vertical lobes. Here, we describe a substructure of the cockroach mushroom bodies composed of a previously unrecognized class of Kenyon cells with distinct morphologies. The embryonically produced class III Kenyon cells form a separate accessory calyx below the calyx proper. The medial branches of class III Kenyon cell axons form the previously described "gamma bulb," whereas the vertical branches leave the vertical lobe to form a toroidal "lobelet" around the posterior surface. Taking advantage of the morphologically and immunochemically distinct nature of the lobelet, we have attempted to determine the distribution of this unique structure in other insects of the taxon Dictyoptera (cockroaches, mantises, and termites). Our data indicate that the lobelet is present only in basal cockroaches and in termites, supporting existing theories of a close phylogenetic relationship between these groups. Higher termites possess a duplicated lobe structure due to immense elaboration of the processes of class III Kenyon cells. The degree of complexity in the mushroom body lobes of termites agrees with current taxonomic arrangements of the Isoptera based on non-neural morphological and DNA sequence analyses. It thus appears that the evolution of the Dictyoptera has been accompanied by increasing complexity of the mushroom bodies, achieved in part through the further specialization and elaboration of a subset of Kenyon cells. PMID- 12532402 TI - Synaptic activity modifies the levels of Dorsal and Cactus at the neuromuscular junction of Drosophila. AB - The Drosophila Rel transcription factor Dorsal and its inhibitor Cactus participate in a signal transduction pathway involved in several biologic processes, including embryonic pattern formation, immunity, and muscle development. In contrast with embryonic muscle, where Dorsal is reportedly absent, this protein and Cactus accumulates in the neuromuscular junctions in the muscle of both larvae and adults. The phenotype of homozygous dorsal mutant larvae suggested that Dorsal and Cactus maybe necessary for normal function and maintenance of the neuromuscular system. Here we investigate if these proteins can respond to synaptic activity. Using larval body wall preparations and antibodies specific for Dorsal or Cactus we show that the amount of these proteins at the neuromuscular junction is substantially decreased after electrical stimulation of the nerves or incubation in glutamate, the principal transmitter in this type of synapse. The specificity of the response was tested with a glutamate receptor antagonist (argiotoxin 636). Because the effect can be reproduced using a calcium ionophore (ionomycin treatment) as well as blocked by the inhibition of the muscle ryanodine receptor (tetracaine treatment), the involvement of calcium in this process seems likely. We also observed that the inhibition of the calcium dependent protein phosphatase calcineurin prevents the effect of glutamate on the fluorescence for Dorsal and Cactus, suggesting its participation in a signal transduction cascade that may activate Dorsal in the muscle independently of Toll. Our results are consistent with a novel function of the Rel factor Dorsal in a molecular pathway turned on by neural activity and/or contractile activity. PMID- 12532405 TI - Central projections of thoracic splanchnic and somatic nerves and the location of sympathetic preganglionic neurons in Xenopus laevis. AB - The central and peripheral organization of thoracic visceral and somatic nervous elements was studied by applying dextran amines to the proximal cut ends of the thoracic splanchnic and somatic nerves in Xenopus laevis. Many labeled dorsal root ganglion cells of visceral afferents, and all somatic afferents, were located in a single ganglion of one spinal segment, and the two types of cells were distributed topographically within the ganglion. The labeled sympathetic preganglionic neurons were located predominantly in the same area of the thoracic spinal gray as in other frogs and in mammals. The labeled visceral afferents projected to Lissauer's tract and the dorsal funiculus. The visceral fibers of the tract ascended to the level of the subcerebellar area, supplying collateral branches to the lateral one-third of the dorsal horn and to the area of brainstem nuclei, including lateral cervical and descending trigeminal nucleus, and descended to the filum terminale. The visceral fibers of the dorsal funiculus were distributed to the dorsal column nucleus and the solitary tract. A similar longitudinal projection was also seen in the somatic afferents. The dual central pathway of thoracic primary afferents in the anuran spinal cord is a property held in common with mammals, but the widespread rostrocaudal projection through Lissauer's tract may be a characteristic of the anuran central nervous system. In frogs, the direct transmission of primary afferent information to an extremely wide area of the central nervous system may be important for prompt assessment of environmental factors and control of body functions. PMID- 12532406 TI - Tenascin-R-deficient mice show structural alterations of symmetric perisomatic synapses in the CA1 region of the hippocampus. AB - Accumulating evidence suggests that extracellular matrix (ECM) molecules play important roles in formation of synapses. Our previous electrophysiologic study of mice deficient in the extracellular matrix glycoprotein tenascin-R (TN-R) showed an impaired gamma-aminobutyric acid release at perisomatic inhibitory synapses in the CA1 pyramidal cell layer of the hippocampus. The present study investigated possible ultrastructural correlates of abnormal perisomatic inhibition. Topographic, morphometric, and stereologic methods were applied at the light and electron microscopic levels to quantify the density and spatial arrangement of cell bodies of CA1 pyramidal neurons and density and architecture of symmetric synapses formed on them in TN-R(-/-) and wild-type mice of different ages. The spatial arrangement of neuronal cell bodies in the CA1 pyramidal cell layer was found more diffuse and disordered in TN-R(-/-) mice than in wild-type animals. The coverage of the plasma membrane of pyramidal cell bodies by active zones of symmetric synapses was reduced by at least 40% in TN-R(-/-) animals compared with control animals. Further, the length of active zone profiles of perisomatic inhibitory synapses in the CA1 pyramidal cell layer was 8-14% smaller, whereas the number of active zones calculated per length unit of cell body profile was 30-40% smaller in TN-R mutants than in wild-type animals. The density and spatial arrangement of synaptic vesicles in the synaptic terminals provided ultrastructural evidence for reduced synaptic activity in TN-R mutants. Thus, TN-R appears to play an important role in the regulation of the number and architecture of perisomatic inhibitory synapses, which play crucial roles in the synchronization of neuronal activity and modulation of synaptic plasticity in the hippocampus. PMID- 12532407 TI - Increased density of hippocampal kainate receptors but normal density of NMDA and AMPA receptors in a rat model of prenatal protein malnutrition. AB - The postnatal development of excitatory amino acid receptor types including kainate, N-methyl-D-aspartate (NMDA), and alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) was assessed in the hippocampus, entorhinal cortex, and adjacent neocortex in normal and prenatally protein malnourished rats ages 15, 30, 90, and 220 postnatal days by quantitative autoradiography. Tritiated ligands used to measure binding site density were (3)[H]kainate, (3)[H]MK-801, and (3)[H]AMPA, respectively. Kainate receptors showed statistically significant increases in binding density in stratum lucidum of CA3 (hippocampal mossy fiber zone) in 90- and 220-day-old malnourished rats compared with age- and sex-matched controls but not in 15- or 30-day-old malnourished rats. Compared with previous anatomic studies, these results are mostly in agreement with a significantly decreased hippocampal mossy fiber plexus in 15-, 90-, and 220-day-old rats but not in 30-day-old rats. These results suggested that the increased density of postsynaptic kainate receptors located mainly on proximal apical dendrites of CA3 pyramidal cells may be compensatory to decreased glutamate release due to the reduction in mossy fiber plexus. In contrast, the density of putative NMDA and AMPA receptors quantified in prenatally malnourished rats was comparable to the density quantified in age- and sex-matched control rats, as were all three receptor types in entorhinal cortex and adjacent neocortex. Thus, the selectivity of the compensation of (3)[H]kainate-labeled mossy fiber plexus in adult but not in early postnatal developing malnourished rats may help ensure continued breeding and survival of the species under otherwise adverse environmental conditions. PMID- 12532408 TI - GABAergic and glutamatergic axons innervate the axon initial segment and organize GABA(A) receptor clusters of cultured hippocampal pyramidal cells. AB - We have studied gamma-aminobutyric acid (GABA)(A) receptor (GABA(A)R) clustering within the axon initial segment (AIS) in low-density cultures of hippocampal pyramidal cells following GABAergic and glutamatergic innervation of the AIS. Large, intensely fluorescent, and postsynaptic GABA(A)R clusters were present in the AIS. More than 95% of these clusters colocalized with presynaptic GABAergic or glutamatergic terminals, forming matched or mismatched synapses, respectively. Less than 5% of the GABA(A)R clusters of the AIS did not colocalize with GABAergic or glutamatergic terminals, suggesting that GABA(A)Rs normally do not form clusters unless the AIS received GABAergic or glutamatergic innervation. Few or no clusters of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA) receptors or the postsynaptic density-95 protein (PSD-95) were found in the AIS, even when the AIS was innervated by glutamatergic axons. Glutamatergic innervation of the AIS that formed mismatched synapses with postsynaptic GABA(A)R clusters mainly occurred when the AIS did not receive GABAergic innervation. However, when the AIS was innervated by GABAergic axons, the formation of matched GABAergic synapses predominated and coincided with large reductions in both the density of glutamatergic terminals from the AIS and the mismatching of GABA(A)R clusters. A similar effect was observed at axo-dendritic synapses, where GABAergic innervation also led to a large decrease in mismatched GABA(A)R clusters and a smaller, but significant, decrease in glutamatergic terminal density in dendrites that received GABAergic innervation. We hypothesize that competition between GABAergic and glutamatergic innervation of the AIS in the intact hippocampus leads to the exclusive presence of GABAergic inhibitory synapses in the AIS of pyramidal cells. PMID- 12532409 TI - PDAPP; YFP double transgenic mice: a tool to study amyloid-beta associated changes in axonal, dendritic, and synaptic structures. AB - Neuritic plaques are one of the stereotypical hallmarks of Alzheimer's disease (AD) pathology. These structures are composed of extracellular accumulations of fibrillar forms of the amyloid-beta peptide (Abeta), a variety of other plaque associated proteins, activated glial cells, and degenerating nerve processes. To study the neuritic toxicity of different structural forms of Abeta in the context of regional connectivity and the entire cell, we crossed PDAPP transgenic (Tg) mice, a model with AD-like pathology, to Tg mice that stably express yellow fluorescent protein (YFP) in a subset of neurons in the brain. In PDAPP; YFP double Tg mice, markedly enlarged YFP-labeled axonal and dendritic varicosities were associated with fibrillar Abeta deposits. These varicosities were absent in areas where there were nonfibrillar Abeta deposits. Interestingly, YFP-labeled varicosities revealed changes that corresponded with changes seen with electron microscopy and the de Olmos silver staining technique. Other silver staining methods and immunohistochemical localization of phosphorylated neurofilaments or phosphorylated tau were unable to detect the majority of these dystrophic neurites. Some but not all synaptic vesicle markers accumulated abnormally in YFP labeled varicosities associated with neuritic plaques. In addition to the characterization of the effects of Abeta on axonal and dendritic structure, YFP labeled neurons in Tg mice should prove to be a valuable tool to interpret the localization patterns of other markers and for future studies examining the dynamics of axons and dendrites in a variety of disease conditions in living tissue both in vitro and in vivo. PMID- 12532410 TI - Expression of PTPRO during mouse development suggests involvement in axonogenesis and differentiation of NT-3 and NGF-dependent neurons. AB - Competition and cooperation between type II and type III receptor protein tyrosine phosphatases (RPTPs) regulate axon extension and pathfinding in Drosophila. The first step to investigate whether RPTPs influence axon growth in the more complex vertebrate nervous system is to identify which neurons express a particular RPTP. We studied the expression of mouse PTPRO, a type III RPTP with an extracellular region containing eight fibronectin type III domains, during embryogenesis and after birth. Mouse PTPRO mRNA is expressed exclusively in two cell types: neurons and kidney podocytes. Maximal expression in the brain was coincident with mid to late gestation and axonogenesis in the brain. We cloned two cDNAs, including a splice variant without sequence coding of 28 amino acids within the juxtamembrane domain that was found mostly in kidney. In situ hybridization detected mPTPRO mRNA in the cerebral cortex, olfactory bulb and nucleus, hippocampus, motor neurons, and the spinal cord midline. In addition, mPTPRO mRNA was found throughout dorsal root, cranial, and sympathetic ganglia and within kidney glomeruli. Mouse PTPRO mRNA was observed in neuron populations expressing TrkA, the high-affinity nerve growth factor receptor, or TrkC, the neurotrophin-3 receptor, and immunoreactive mPTPRO and TrkC colocalized in large dorsal root ganglia proprioceptive neurons. Our results suggest that mPTPRO is involved in the differentiation and axonogenesis of central and peripheral nervous system neurons, where it is in a position to modulate intracellular responses to neurotrophin-3 and/or nerve growth factor. PMID- 12532411 TI - Distribution of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate-type glutamate receptor subunits (GluR2/3) along the ventral visual pathway in the monkey. AB - By using immunohistochemical methods, we examined the distribution of cells expressing subunits of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)-selective glutamate receptors (GluR2/3) in the cortical areas of the occipitotemporal pathway in monkeys. GluR2/3-immunoreactive (-ir) cells were primarily pyramidal cells; this category, however, also included large stellate cells in layer IVB of the striate cortex (V1) and fusiform cells in layer VI of all the areas examined. GluR2/3 immunoreactivity differed among the areas in laminar distribution and intensity. In V1, GluR2/3-ir cells were identified mainly in layers II, III, IVB, and VI. The prestriate areas V2 and V4 and the inferior temporal areas TEO and TE contained GluR2/3-ir cells in layers II, III, and VI. In the TE, GluR2/3-ir cells were also abundant in layer V. In area 36 of the perirhinal cortex, neurons in layers II, III, V, and VI were labeled in a similar manner to the TE labeling, but with greater staining intensity and numbers, especially in layer V. Thus, GluR2/3 immunoreactivity increased rostrally along the pathway. Within V1 and V2, cells strongly stained for GluR2/3 formed clusters that colocalized with cytochrome oxidase (CO)-rich regions. These distinct laminar and regional distribution patterns of GluR2/3 expression may contribute to the specific physiological properties of neurons within various visual areas and compartments. PMID- 12532412 TI - Very low concentrations of arsenite suppress poly(ADP-ribosyl)ation in mammalian cells. AB - Arsenite is a naturally occurring environmental pollutant of major concern, since adverse health effects including cancer of skin and internal organs have been attributed to chronic arsenic exposure especially via drinking water. Arsenite is not a significant inducer of point mutations but exerts clastogenic activities and interferes with various DNA repair systems at concentrations in the low micromolar range. Nevertheless, no single DNA repair protein exquisitely sensitive to arsenic has been identified. Here we report that poly(ADP ribosyl)ation, which is predominantly mediated by poly(ADP-ribose) polymerase-1 (PARP-1), is inhibited at concentrations as low as 10 nM in cultured HeLa cells, closely matching arsenic concentrations in blood and urine of the general population. Since poly(ADP-ribosyl)ation is an immediate cellular response to DNA damage, playing a major role in DNA base excision repair and the maintenance of genomic stability, its inhibition by arsenite may add to the risk of cancer formation under low-exposure conditions. PMID- 12532413 TI - Deguelin suppresses the formation of carcinogen-induced aberrant crypt foci in the colon of CF-1 mice. AB - Deguelin [(7aS,BaS)-13,13a-dihydro-9,10-dimethoxy-3,3-dimethyl-3H Bis[1]benzopyrano[3,4-b:6',5'-e]pyran-7(7aH)-one], a naturally occurring rotenone, has shown chemopreventive efficacy in several in vivo and in vitro models. In this report, the effectiveness of deguelin at inhibiting the development of AOM-induced colonic ACF was investigated in CF-1 mice. Loss of hex activity was assessed as a second biomarker. In an initial experiment, animals were given s.c. injections of AOM (10 mg/kg body weight) once a week for 2 weeks to induce ACF. Deguelin and vehicle (corn oil) were administered i.g. 7 days a week. Treatment was initiated 2 weeks prior to the first dose of carcinogen and continued for the duration of the study. The mean number of ACF for the control group was 29.0 +/- 4.3, whereas the mean numbers of ACF in the deguelin groups were 24.8 +/- 2.7, 7.2 +/- 1.5 and 4.6 +/- 1.4 at doses of 2.5, 5.0 and 10.0 mg/kg body weight, respectively. In a similar manner, treatment with deguelin significantly (p < 0.001) suppressed the appearance of hex(-) crypts in a dose dependent manner. In a second study, the ability of deguelin to block the initiation and promotion stages of colon carcinogenesis was investigated. Greatest inhibition was observed when deguelin was administered during the promotional stage (73.3%, p < 0.001). These results demonstrate that deguelin is an efficacious chemopreventive agent against colon carcinogenesis. PMID- 12532414 TI - Cannabinoids inhibit gap junctional intercellular communication and activate ERK in a rat liver epithelial cell line. AB - Many tumor promoters suppress the immune system; however, the direct effect of immunosuppressants on the tumorigenic pathways of nonimmune cells in solid tissue has not been well documented. Cannabinoids were chosen to explore this question further. Cannabinoids are immune modulators that affect specific intracellular signaling pathways in leukocytes. Since these compounds are nongenotoxic, any tumorigenic effect that might be associated with these compounds would need to occur through an epigenetic mechanism. Therefore, we determined the effect of Delta(9)-THC and CBN, 2 plant-derived cannabinoids, on 2 key epigenetic markers of tumor promotion: inhibition of GJIC, which is essential in removing a cell from growth suppression, and activation of the ERK-MAPK pathway, which is crucial in activating the appropriate genes for mitogenesis. Both Delta(9)-THC and CBN reversibly inhibited GJIC at noncytotoxic doses (15 microM) in a normal diploid WB rat liver epithelial oval cell line within 20 min and activated ERK1 and ERK2 within 5 min. Inhibition of MEK with PD98059 prevented the inhibition of GJIC by either cannabinoid, suggesting that inhibition of GJIC was MEK-dependent. Based on RT-PCR analysis and employment of an antagonist of CB1 and CB2, the effects on GJIC and MAPK were independent of both cannabinoid receptors. Cannabinoids affected crucial epigenetic pathways associated with cell proliferation in a rodent liver epithelial cell model system. PMID- 12532415 TI - Spontaneous activation and signaling by overexpressed epidermal growth factor receptors in glioblastoma cells. AB - Overexpressed epidermal growth receptor factor receptors (EGFRs) are thought to contribute to the malignant phenotype of human glioblastomas (GBMs), but the mechanism is not well understood. We found that SKMG-3 cells, a rare GBM cell line that maintains EGFR gene amplification in vitro, produced high levels of EGFR protein. The cells also expressed the related receptors HER2/neu and HER4, but not HER3. Immunoblots and tryptic phosphopeptide maps showed that the SKMG-3 EGFRs were intact and functional and that a subset of these receptors were spontaneously autophosphorylated. EGF treatment stimulated phosphorylation of the EGFRs as well as the downstream effectors Erk, AKT1, stat3 and c-Cbl. Under minimal growth conditions, the unstimulated SKMG-3 cells contained constitutively phosphorylated Erk and AKTI but no detectable stat3 DNA-binding complexes. The EGFR kinase inhibitor PD158780 reduced the constitutive phosphorylation of the receptor and Erk but not that of AKT1. In contrast, inhibition of phosphatidylinositol-3-kinase (PI3K) blocked the constitutive phosphorylation of Erk and AKT-1 but not the EGFR. We conclude that the SKMG-3 cells represent the subset of GBMs with amplified EGFR genes that overexpress intact receptors. The results also suggest that in some GBMs, signals from overexpressed EGFRs contribute to the constitutive phosphorylation of Erk, but these signals may not required for the constitutive activation of PI3K or AKT1. PMID- 12532416 TI - Age-dependent variations of human and rat colon myofibroblasts in culture: Influence on their functional interactions with colon cancer cells. AB - Epithelial-mesenchymal interactions play a pivotal role in colon cancer invasion and metastasis. We aimed at elucidating the impact of long-term cultivation on the phenotypic and functional characteristics of primary fibroblasts and their interaction with the human colon adenocarcinoma cell line LoVoC5. We used fibroblasts from human colon tumor tissue, normal human colon mucosa, rat normal colon and 2 rat colon-derived myofibroblast cell lines, MIC316 and MG. The following parameters were studied: cell shape and size, growth curve, intermediate filament expression and extracellular matrix synthesis. Coculture models with or without cell contacts were used to test the effects on LoVoC5 cell proliferation, spreading and adhesion. Irrespective of their origin, fibroblastic cells in primary cultures presented marked phenotypic and functional changes with time. Before passage 5, they presented as large, slow-growing cells expressing vimentin and alpha-smooth muscle actin; synthesizing laminin-1, fibronectin and collagens I and IV; and inducing LoVoC5 proliferation, spreading and adhesion. After passage 15, they presented as small, fast-growing cells inconstantly expressing alpha-smooth muscle actin and synthesizing mainly type I collagen. In coculture with or without cell contacts, they inhibited LoVoC5 proliferation and allowed only limited cell spreading and adhesion. Myofibroblastic cell lines presented as large, fast-growing cells expressing vimentin and alpha-smooth muscle actin and synthesizing mainly type I collagen. They had no significant effects on LoVoC5 proliferation, spreading and adhesion. Our results underline the importance of age-dependent variations in colon mesenchymal cells in culture and for the in vitro study of epithelial-mesenchymal interactions in colon cancer. PMID- 12532417 TI - Development of resistance to vincristine and doxorubicin in neuroblastoma alters malignant properties and induces additional karyotype changes: a preclinical model. AB - Cytotoxic drug treatment of neuroblastoma often leads to the development of drug resistance and may be associated with increased malignancy. To study the effects of long-term cytotoxic treatment on malignant properties of tumor cells, we established 2 neuroblastoma cell sublines resistant to vincristine (VCR) and doxorubicin (DOX). Both established cell lines (UKF-NB-2(r)VCR(20) and UKF-NB 2(r)DOX(100)) were highly resistant to VCR, DOX and vice-versa but retained their sensitivity to cisplatin. UKF-NB-2(r)VCR(20) and UKF-NB-2(r)DOX(100) expressed significant amounts of P-glycoprotein, while parental cells were P-glycoprotein negative. GD2 expression was upregulated, whereas NCAM expression was decreased in both resistant cells. Spectral karyotype (SKY) analysis revealed complex aberrant karyotypes in all cell lines and additional acquired karyotype changes in both resistant cells. All cell lines harbored high levels of N-myc amplification. Compared to parental cells, UKF-NB-2(r)VCR(20) and UKF-NB 2(r)DOX(100) exhibited more than 2-fold increase in clonal growth in vitro, accelerated adhesion and transendothelial penetration and higher tumorigenicity in vivo. We conclude that development of drug resistance and acquisition of certain karyotypic alterations is associated with an increase of additional malignant properties that may contribute to the poor prognosis in advanced forms of NB. The 2 novel neuroblastoma cell sublines also provide useful models for the study of drug resistance in aggressive forms of neuroblastoma. PMID- 12532418 TI - Effects of E-cadherin transfection on gene expression of a gallbladder carcinoma cell line: repression of MTS1/S100A4 gene expression. AB - E-cadherin is important in cell-to-cell adhesion and controls cell polarity and tissue morphology. Loss of E-cadherin expression occurs in various human tumors and is the first step in cancer invasion and metastasis. We demonstrate that the exogenous expression of E-cadherin transfected into G-415 GB cells not only increases cell-to-cell adhesion but also reduces in vitro cell proliferation, motility and invasion. Our aim was to determine what genes are most affected by the exogenous expression of E-cadherin in GB cancer cells. We analyzed gene expression pertaining to cell proliferation, motility and invasion. Conventional RT-PCR was performed for these genes; quantitative RT-PCR was carried out on genes exhibiting altered expression. Conventional RT-PCR revealed that E-cadherin transfection suppressed expression of mts1 mRNA and increased that of c-myc and MT1-MMP. In quantitative RT-PCR analysis, levels of c-myc and MT1-MMP mRNA were elevated by to 2.56- and 2.22-fold, respectively, in the E-cadherin transfectant, whereas mts-1 was 7.14-fold suppressed compared to parental cells. These results indicated that expression of mts1 mRNA was most affected by E-cadherin transfection. Immunocytochemical analysis of transfectant and parental cells demonstrated an inverse correlation in E-cadherin and mts1 expression. Immunohistochemical analysis of 37 GB cancer specimens confirmed this observation in vivo. Loss of E-cadherin expression followed by expression of the mts1 gene may be an important event for increasing cell proliferation, motility and invasion activity in the progression of GB cancer. PMID- 12532419 TI - 8q24 Copy number gains and expression of the c-myc mRNA stabilizing protein CRD BP in primary breast carcinomas. AB - The coding region determinant binding protein (CRD-BP) was isolated by virtue of its high affinity to the c-myc mRNA coding region stability determinant and shown to shield this message from nucleolytic attack, prolonging its half-life. CRD-BP is normally expressed during fetal life but is also activated de novo in tumors. Considering that aberrant CRD-BP expression may represent an additional mechanism interfering with c-myc regulation, we screened 118 primary breast carcinomas for CRD-BP expression, 60 of which had also been analyzed by comparative genomic hybridization (CGH). Copy number gains encompassing 8q24, the chromosome band that contains the c-myc locus, were detected in 48.3% (29/60) of tumors, whereas gains involving band 17q21, which contains the CRD-BP locus, were observed in 18.3% (11/60) of tumors. CRD-BP expression was detected in 58.5% (69/118) of tumors, implying mechanisms of activation alternative to gene amplification. Altogether, some 75% of the tumors had alterations pertaining to c-myc since they either harbored 8q24 gains and/or expressed CRD-BP. Significant associations were detected between CRD-BP expression and the absence of estrogen receptors (p = 0.005) and between the presence of 8q24 gains and an increased number of genomic changes as measured by CGH (p = 0.0017). Tumors were divided into 4 groups according to CRD-BP expression and 8q24 gains. The odds for tumors having both characteristics to be classified as poorly differentiated (grade III vs. grade I and II) were 19.6 times the corresponding odds for tumors neither expressing CRD BP nor harboring 8q24 gains. For tumors either harboring 8q24 gains only or expressing CRD-BP alone, the corresponding odds were 6.4 and 3, respectively. PMID- 12532420 TI - Relationship between E-cadherin gene mutation and p53 gene mutation, p53 accumulation, Bcl-2 expression and Ki-67 staining in diffuse-type gastric carcinoma. AB - E-cadherin mutations are found in 50% of diffuse-type gastric carcinoma, but not in intestinal gastric carcinoma. Because cell-cell adhesion mediated by E cadherin plays an important role in epithelial cell survival, E-cadherin mutations could alter the apoptotic behavior of tumor cells. p53 and Bcl-2 family members are also important regulators of cellular apoptosis. This is the first study that investigates the relationship between E-cadherin gene mutation and p53 gene mutation, p53 accumulation, Bcl-2 expression, and Ki-67 expression in diffuse-type gastric carcinoma (24 cases, E-cadherin mutation status: wild-type in 8 patients and mutant in 16 patients). The mutation status of exons 5-8 of p53 was analyzed by denaturing high pressure liquid chromatography (DHPLC) in formalin-fixed, paraffin-embedded tumor sections, followed by direct sequencing of cases with aberrant chromatographic patterns. p53 mutations were found in 1 of 8 tumors without E-cadherin mutation (12.5%) and in 1 of 16 tumors with E cadherin mutation (6.3%), a difference that was not statistically significant (p = 1.00). p53 accumulation was found in 8 of 24 tumors (33.3%) by immunohistochemical staining. p53 accumulation was significantly more frequent in tumors without E-cadherin mutations (5 of 8 tumors, 62.5%) than in gastric carcinoma tissues with E-cadherin mutations (3 of 16 tumors, 18.8%, p = 0.03). Bcl-2 staining was not observed in gastric carcinoma cells without E-cadherin mutations, but was detectable in 5 of 16 tumors with E-cadherin mutations (31.3%), a difference that was not statistically significant (p = 0.13). No relationship was observed between Ki-67 staining and the E-cadherin mutation status (p = 1.00). These data suggest that the presence of E-cadherin mutations can significantly alter the accumulation of the apoptosis-regulating p53 protein, whereas no correlation with the p53 mutation status or with Ki-67 staining was observed. PMID- 12532421 TI - Molecular basis of basal cell carcinoma: analysis of differential gene expression by differential display PCR and expression array. AB - Basal cell carcinoma (BCC) is the most common tumor in the Caucasian population. Although BCC rarely metastasize and cause death, they are problematic due to their destructive growth and the frequent localization on the face. Until now the knowledge of genes differentially expressed in BCC has been incomplete. To elucidate the complex alterations in BCC-associated gene expression, we took advantage of 2 techniques: the differential display RT-PCR (DD-PCR) and the differential hybridization of cDNA arrays. Using DD-PCR, we showed differential expression of genes known from other biological contexts (e.g., rac, ubiquitin hydrolase), which could now be associated with BCC. In addition, we detected unknown genes possibly contributing to the carcinogenesis of BCC. Of the 588 genes screened by differential hybridization of the Atlas human cDNA array, differences in the expression levels of BCC were observed for 10 genes. These data were obtained with RNA probes pooled from several BCC of different donors and were subsequently confirmed by semiquantitative RT-PCR for Janus protein tyrosine kinase 3 (Jak3), microsomal glutathione S-transferase 1 (GST 12), teratocarcinoma-derived growth factor cripto, glutaredoxin and the monocyte chemoattractant protein 1 (MCP-1) in 10 individual BCC specimens, 2 squamous cell carcinoma (SCC), the cell line HaCaT and cultured normal human keratinocytes (NHK) in comparison to normal skin. These genes are candidates from gene families with known association to tumors, but they have not been reported in the carcinogenesis of BCC yet. In summary, both approaches allow the detection of differentially expressed genes possibly involved in the carcinogenesis of BCC. PMID- 12532422 TI - Serologic analysis of ovarian tumor antigens reveals a bias toward antigens encoded on 17q. AB - We utilized SEREX immunoscreening to identify a set of novel tumor antigens that are associated with human serous ovarian cancer and may prove useful for the early detection and treatment of this disease. Extensive screening with a panel of sera from 25 late-stage ovarian cancer patients against 3 independent cDNA libraries identified a set of 9 antigens that were immunogenic in more than 1 patient and not in a panel of 20-45 normal female serum donors. These antigens include p53, NY-ESO-1, UBQLN1, HOXB6, TOP2A, putative helicase-RUVBL (RUVBL), HMBA-inducible (HEXIM1), DDX5 and HDCMA. Ten of 25 ovarian cancer patients (40%) expressed serum IgG to at least 1 of these antigens, while 14% (4/25) had antibodies to 2 or more antigens. Unexpectedly, 4 antigens identified in this screen, DDX5, HEXIM1, TOP2A and HOXB6, are encoded within a region of 17q that also includes the genes for HER2/neu, Homeobox-B7 and BRCA1. Real-time RT-PCR analysis showed that mRNA for HER2/neu and 3 SEREX-defined antigens, TOP2A, HOXB6 and DDX5, was more abundant in ovarian tumors than most normal tissues, including normal and benign ovarian tissues, suggesting that elevated expression of genes encoded within this region of chromosome 17 is a common event in ovarian tumors. Thus, these abnormal expression patterns combined with the endogenous immune response suggests that these antigens represent potential targets for immunotherapy. PMID- 12532424 TI - Prognostic value of tumor-infiltrating dendritic cells expressing CD83 in human breast carcinomas. AB - DCs are the most potent antigen-presenting cells that play a major role in initiating the antitumor immune response. Although the clinical significance of TIDCs has been investigated in a variety of human cancers, few studies have focused on the in situ maturation status of DCs. We have analyzed the maturation specific significance of TIDCs in the prognosis of patients with breast carcinoma. We evaluated 130 breast carcinomas for the presence of TIDCs using immunohistochemistry with an anti-CD1a antibody for immature DCs and an anti-CD83 antibody for mature DCs. Intratumoral expression of immunosuppressive cytokines was also examined. All samples contained CD1a(+) TIDCs, and 82 (63.1%) samples contained CD83(+) TIDCs. The number of CD83(+) TIDCs was inversely correlated with lymph node metastasis and with tissue expression of VEGF and TGF-beta, whereas the number of CD1a(+) TIDCs was not. Kaplan-Meier analysis (log rank statistics) revealed a significant association of increasing number of CD83(+) TIDCs with longer relapse-free (p = 0.002) and overall (p < 0.001) survival. Furthermore, among patients with lymph node metastasis, the survival rate of those with larger numbers of CD83(+) TIDCs was significantly better than that of patients with fewer CD83(+) TIDCs. Multivariate analysis revealed that CD83(+) TIDCs had independent prognostic relevance in breast carcinomas. The infiltration of tumors by mature DCs expressing CD83 may be of great importance in initiating the primary antitumor immune response and is confirmed as an independent, immunologic prognostic parameter for survival in patients with breast cancer. PMID- 12532423 TI - GLUT-1 and CAIX as intrinsic markers of hypoxia in carcinoma of the cervix: relationship to pimonidazole binding. AB - The presence of hypoxia in tumours results in the overexpression of certain genes, which are controlled via the transcription factor HIF-1. Hypoxic cells are known to be radioresistant and chemoresistant, thus, a reliable surrogate marker of hypoxia is desirable to ensure that treatment may be rationally applied. Recently, the HIF-1-regulated proteins Glut-1 and CAIX were validated as intrinsic markers of hypoxia by comparison with pO(2) measured using oxygen electrodes. We compare the expression of Glut-1 and CAIX with the binding of the bioreductive drug hypoxia marker pimonidazole. Pimonidazole was administered to 42 patients with advanced carcinoma of the cervix, 16 hr before biopsy. Sections of single or multiple biopsies were then immunostained for Glut-1 and CAIX, and the area of staining scored by eye, using a "field-by-field" semi-quantitative averaging system. Using 1 biopsy only, Glut-1 (r = 0.54, p = <0.001) correlated with the level of pimonidazole binding, and Glut-1 and CAIX expression also correlated significantly (r = 0.40, p = <0.009). Thus, our study has shown that HIF-1 regulated genes have potential for future use as predictors of the malignant changes mediated by hypoxia, and warrant further investigation as indicators of response to cancer therapy. PMID- 12532425 TI - Clinical course of bladder neoplasms and single nucleotide polymorphisms in the CDKN2A gene. AB - Point mutations and single nucleotide polymorphisms (SNPs) in the CDKN2A gene in bladder cancer patients have been resolved only to a limited extent. The exact frequency of mutations remains uncertain and reports on SNPs are lacking. In this population-based study we investigated mutations and polymorphisms in the CDKN2A gene in bladder cancer patients from all hospitals within the Stockholm County. Mutations were determined in 4 exons of the CDKN2A gene in tumor-tissues from 172 bladder cancer patients and 2 single nucleotide polymorphisms in the 3' UTR of the CDKN2A gene were studied in 309 cases. Missense mutations were identified in only 4 of 172 (2.3%) cases, including 1 in the germ-line. Frequencies of the 500 C-->G and 540 C-->T polymorphisms in the 3' UTR of the CDKN2A in bladder cancer cases were not statistically significantly different compared to an ethnically matched control population. The tumor-specific survival was significantly shorter in patients with either the 500 C-->G or 540 C-->T polymorphism than those with wild-type CDKN2A gene (P = 0.02). Our results corroborate the earlier findings that single base mutation is not the prime mode of inactivation of the CDKN2A gene in bladder cancer. Further, the results indicate, a role for the 3' UTR polymorphisms in the CDKN2A gene in tumor invasiveness. PMID- 12532426 TI - Immune enhancement of nitroreductase-induced cytotoxicity: studies using a bicistronic adenovirus vector. AB - The nitroreductase (NR)/CB1954 enzyme prodrug system has given promising results in preclinical studies and is currently being assessed in phase I clinical trials. It is well established that there is an immune component to the bystander effect observed with other systems such as thymidine kinase and cytosine deaminase; however, such an effect has not previously been described using NR. We have preliminary data suggesting an immune bystander effect with NR to further examine these effects and their potential enhancement by cytokines, an adenoviral vector containing CMV-NR, an internal ribosome entry site (IRES) and the gene for murine GM-CSF (mGM-CSF) was constructed. The NR-GM-CSF virus was validated in 2 experimental models and demonstrated increased therapeutic efficacy in the MC26 murine colorectal tumour model. These data illustrate that the combination of suicide gene therapy using NR and CB1954 with immune stimulation via GM-CSF gives an improved response compared to either modality alone and suggests that the immune component of this response may be beneficial in combating unresectable, metastatic disease and preventing tumour recurrence. PMID- 12532427 TI - Dynamic T1-weighted monitoring of vascularization in human carcinoma heterotransplants by magnetic resonance imaging. AB - Studies on tumor angiogenesis and antiangiogenic therapies are commonly performed with tumor heterotransplants in nude mice. To monitor therapeutic effects, improved noninvasive analyses of functional data are required, in addition to the assessment of tumor volume and histology. Here, we report on sequential monitoring of vascularization of human squamous cell carcinomas growing as heterotransplants in nude mice using MRI. Using a custom-developed animal coil in a conventional whole-body 1.5 T MRI scanner, dynamic T1w sequences were recorded after i.v. injection of Gd-DTPA in tumors grown for 17, 21, 25, 29 and 33 days. Amplitude and the exchange rate constant (k(ep)) were calculated according to a 2 compartment model, discriminating intravascular and interstitial spaces, and correlated with tumor size and histology. High-resolution imaging of small heterotransplants from 100 to 1,000 mm(3) was achieved, clearly discriminating vital and necrotic areas. Preceding the development of necroses, which were hyperintense in T2w images and confirmed with histology, a local decrease of amplitude and k(ep) values was observed. Significantly higher amplitudes were found in tumor periphery than in central parts, correlating well with the vascular pattern obtained by immunocytochemistry. Tumor size correlated negatively with amplitude, probably as a result of increasing necrotic areas, whereas the reason for the observed increase of k(ep) value with tumor size remains unclear. These data demonstrate that dynamic MRI is an excellent method for noninvasive assessment of tumor vascularization in small animals using a clinical whole-body scanner with little technical modifications. This technique provides functional data characterizing essential features of tumor biology and is thus appropriate for monitoring antiangiogenic therapies. PMID- 12532428 TI - Comparison of antiangiogenic activities using paclitaxel (taxol) and docetaxel (taxotere). AB - Tumor growth requires a competent vascular supply and angiogenesis has been considered as a potential target for the treatment of several cancers. The two clinically approved taxanes, paclitaxel and docetaxel, are novel antimitotic agents that are under extensive investigation in clinical trials. Both taxanes have demonstrated significant activity against many solid tumors, but little is known about the effect of paclitaxel and docetaxel on endothelial cell function and angiogenic processes. The purpose of our study was to examine and compare the effects of these drugs on angiogenic processes in vitro and in vivo. These processes include: proliferation, migration and differentiation of cultured human umbilical vein endothelial cells (HUVEC) (in vitro), capillary sprouting of rat aortic ring explants (ex vivo) and HT1080 tumor growth in vivo. Our results demonstrate that endothelial cells are 10-100-fold more sensitive to these drugs than tumor cells. Additionally, comparison of the taxanes demonstrated that angiogenesis is blocked by both drugs primarily via inhibition of proliferation and differentiation (tube assay) and induction of cell death. Docetaxel, however, appears to be more potent at inhibiting angiogenesis, with an IC(50) concentration 10x less than that of paclitaxel. We conclude that these important findings should be taken in account in clinical trials where tumor angiogenesis is being targeted. PMID- 12532430 TI - Current treatment for liver metastases from colorectal cancer. AB - The liver is the commonest site of distant metastasis of colorectal cancer and nearly half of the patients with colorectal cancer ultimately develop liver involved during the course of their diseases. Surgery is the only therapy that offers the possibility of cure for patients with hepatic metastatic diseases. Five-year survival rates after resection of all detectable liver metastases can be up to 40 %. Unfortunately, only 25 % of patients with colorectal liver metastases are candidates for liver resection, while the others are not amenable to surgical resection. Regional therapies such as radiofrequency ablation and cryotherapy may be offered to patients with isolated unresectable metastases but no extrahepatic diseases. Hepatic artery catheter chemotherapy and chemoembolization and portal vein embolization are often used for the patients with extensive liver metastases but without extrahepatic diseases, which are not suitable for regional ablation. For the patients with metastatic colorectal cancer beyond the liver, systemic chemotherapy is a more appropriate choice. Immunotherapy is also a good option when other therapies are used in combination to enhance the efficacy. Selective internal radiation therapy is a new radiation method which can be used in patients given other routine therapies without effects. PMID- 12532431 TI - Hepatic stem cells: existence and origin. AB - Stem cells are not only units of biological organization, responsible for the development and the regeneration of tissue and organ systems, but also are units in evolution by natural selection. It is accepted that there is stem cell potential in the liver. Like most organs in a healthy adult, the liver maintains a perfect balance between cell gain and loss. It has three levels of cells that can respond to loss of hepatocytes: (1) Mature hepatocytes, which proliferate after normal liver tissue renewal, less severe liver damage, etc; they are numerous, unipotent, "committed" and respond rapidly to liver injury. (2) Oval cells, which are activated to proliferate when the liver damage is extensive and chronic, or if proliferation of hepatocytes is inhibited; they lie within or immediately adjacent to the canal of Hering (CoH); they are less numerous, bipotent and respond by longer, but still limited proliferation. (3) Exogenous liver stem cells, which may derive from circulating hematopoietic stem cells (HSCs) or bone marrow stem cells; they respond to allyl alcohol injury or hepatocarcinogenesis; they are multipotent, rare, but have a very long proliferation potential. They make a more significant contribution to regeneration, and even completely restore normal function in a murine model of hereditary tyrosinaemia. How these three stem cell populations integrate to achieve a homeostatic balance remains enigmatic. This review focuses on the location, activation, markers of the three candidates of liver stem cell, and the most importantly, therapeutic potential of hepatic stem cells. PMID- 12532432 TI - Overexpression of ETS2 in human esophageal squamous cell carcinoma. AB - AIM: To study the expression pattern of ETS2 (erythroblastosis virus oncogene homolog 2) in human esophageal squamous cell carcinoma (ESCC). METHODS: Reverse transcription polymerase chain reaction (RT-PCR) and Northern blot were performed to examine the expression level of ETS2 mRNA in 37 pairs of ESCC tissue samples. Western blot and immunohistochemistry were carried out to check the expression level of ETS2 protein in 30 pairs of ESCC tissue specimens. RESULTS: RT-PCR and Northern blot analysis showed that ETS2 mRNA upregulated in 75.7 % (28/37) examined ESCC tissues relative to matched normal tissues. From those 37 cases, 14 cases were randomly selected to perform Western blot and the results revealed that ETS2 protein overexpressed in 71.4 % (10/14) checked ESCC tissues compared with the corresponding normal tissues. Moreover, the expression patterns of ETS2 protein in those 14 cases were identical to those of ETS2 mRNA displayed by RT PCR or Northern Blot. Immunohistochemistry analysis showed that the expression level of ETS2 protein rose in 75 % (12/16) tumor epithelial cells contrasted to the normal cells. Altogether the expression level of ETS2 protein increased in 73.3 % (22/30) checked ESCC tissue samples contrary to their normal counterparts. CONCLUSION: The results suggested that ETS2 overexpressed in paired human ESCC tissue samples at both mRNA and protein levels and may be associated with the tumorigenesis of esophagus. PMID- 12532433 TI - An analysis of esophageal cancer incidence in Cixian county from 1974 to 1996. AB - AIM: To describe the incidence of esophageal cancer (EC) in Cixian, a county of Hebei province during 1974-1996. We analyzed the sex and age characteristics as well as the geographic distribution of EC, in order to determine the impact so that methods of preventing and controlling EC in Cixian can be put in place. METHODS: Since the early 1970s, the cancer registry system has been established, which collects the cancer incidence in Cixian county. The malignant tumors were coded according to International Classification of Disease IX (ICD-9). All the data were checked and analyzed using EPIINFO. RESULTS: The trend of the incidence rate of EC from 1974 to 1996 had declined, (229.9/100 000 vs 178.5/100 000, Odds ratio=1.47, 95 % CI:1.32-1.63, chi(2)=52.89. trend chi(2)=26.54, P<0.001). The incidence rate of males declined significantly (281.81/100 000 vs 157.96/100 000, Odds ratio=1.61, 95 % CI: 1.41-1.84, chi(2)=47.85. Trend chi(2)=44.86, P<0.001), whereas, the females remained steady (157.96/100 000 vs 133.41/100 000, odds ratio=1.28, 95 % CI:1.17-1.49, chi(2)=9.26. trend chi(2)=2.69, P>0.05). Male average annual incidence rate was 142.80/100 000 and the female's was 95.18/100 000. The sex ratio (males to females) was 1.50:1. The incidence rate was increasing along with the age. As to the geographic distribution, the incidence rate in mountainous areas and hilly areas showed a significantly declining trend (mountainous areas, trend chi(2)=149.93, P<0.001; hilly areas, trend chi(2)=42.70, P<0.001). The incidence rate of EC in plain areas had increased (trend chi(2)=22.39, P<0.001). CONCLUSION: The incidence rate of EC in Cixian county shows a trend and has declined after two decades, especially in mountainous area. But compared to other regions in the world, Cixian county still had a high incidence rate of EC. PMID- 12532434 TI - Risk factors for the gastric cardia cancer: a case-control study in Fujian Province. AB - AIM: The incidence of gastric cardia cancer has greatly increased in the past 2-3 decades, however, the risk factors for the disease are still not clearly understood. The investigations among Chinese population on the risk factors of gastric cardia cancer were also scarcely reported. We therefore conducted a case control study in Fujian province, China, to investigate the potential risk and protective factors of this disease. METHODS: 191 cardia and 190 non-cardia gastric cancer cases, and a total of 222 control cases were included in this study. Standard questionnaires were used in collecting epidemiological factors and the data were then analyzed by the unconditional logistic regression model. RESULTS: As the factors such as age, gender, smoking, alcohol consumption, and family history of gastric cancer were controlled, a multivariable analysis was conducted, which revealed that there was a significant correlation between the dietary habits such as irregular meal, over and fast eating, and the gastric cardia cancer with the odds ratios (ORs) of 4.2 (95 % confidence interval: 2.3 7.7), 4.7 (2.1-10.8), and 2.7 (1.3-5.3) respectively. Other correlations were also observed between the gastric cardia cancer and the consumption of salty fish or pickled vegetable, smoking, and the family cancer history with the ORs of 5.5 (1.4-19.5), 1.8 (1.0-3.0), 2.1 (1.3-3.5), and 3.8 (2.3-6.2) respectively. In contrast, the negative correlations were found existing between the intake of fresh vegetables and fruits, the use of refrigerator, and the gastric cardia cancer, with the ORs of 0.4 (0.2-0.9), 0.2 (0.1-0.5), and 0.2 (0.1-0.4), respectively. However, dietary habits were associated less with non-cardia gastric cancer compared with its cardia counterpart. CONCLUSION: Dietary habits might be one of the risk factors for the cardia carcinogenesis among Chinese population. PMID- 12532435 TI - Preoperative TN staging of esophageal cancer: comparison of miniprobe ultrasonography, spiral CT and MRI. AB - AIM: To evaluate the value of miniprobe sonography (MPS), spiral CT and MR imaging (MRI) in the tumor and regional lymph node staging of esophageal cancer. METHODS: Eight-six patients (56 men and 30 women; age range of 39-73 years, mean 62 years) with esophageal carcinoma were staged preoperatively with imaging modalities. Of them, 81 (94 %) had squamous cell carcinoma, 4(5 %) adenocarcinoma, and 1(1 %) adenoacanthoma. Eleven patients (12 %) had malignancy of the upper one third, 41 (48 %) of the mid-esophagus and 34 (40 %) of the distal one third. Forty-one were examined by spiral CT in whom 13 were co examined by MPS, and forty-five by MRI in whom 18 were also co-examined by MPS. These imaging results were compared with the findings of the histopathologic examination for resected specimens. RESULTS: In staging the depth of tumor growth, MPS was significantly more accurate (84 %) than spiral CT and MRI (68 % and 60 %, respectively, P<0.05). The specificity and sensitivity were 82 % and 85 % for MPS; 60 % and 69 % for spiral CT; and 40 % and 63 % for MRI, respectively. In staging regional lymph nodes, spiral CT was more accurate (78 %) than MPS and MRI (71 % and 64 %, respectively), but the difference was not statistically significant. The specificity and sensitivity were 79 % and 77 % for spiral CT; 75 % and 68 % for MPS; and 68 % and 62 % for MRI, respectively. CONCLUSION: MPS is superior to spiral CT or MRI for T staging, especially in early esophageal cancer. However, the three modalities have the similar accuracy in N staging. Spiral CT or MRI is helpful for the detection of far-distance metastasis in esophageal cancer. PMID- 12532436 TI - Expression of e-cadherin and beta-catenin in human esophageal squamous cell carcinoma: relationships with prognosis. AB - AIM: To elucidate the expression of E-cadherin and beta-catenin correlating with its clinical outcome in patients with esophageal squamous cell carcinoma (ESCC), by analyzing their interrelationship with clinicopathological variables and their effects on progress and prognosis. METHODS: Expression of E-cadherin and beta catenin was determined by SP immunohistochemical technique in patients with ESCC consecutively, their correlation with clinical characteristics was evaluated and analyzed by multivariate analysis. RESULTS: The rate of expression of E-cadherin decreased to 66.03 % (70/106) in ESCC and the protein level was negative correlated with histologic grade, tumor size, clinical staging, lymph node metastasis and venous invasion. Whereas the expression rate of beta-catenin was reduced to 69.8 % (74/106) and the level of protein expression correlated only with histologic grade. There obviously existed inverse correlation between level of E-cadherin protein and survival, especially in stage I, IIa, IIb (P=0.0033), Patients with low-expressing tumors for beta-catenin and non-expressing tumors for E-cadherin/beta-catenin had lower survival period than those with normal expressing ones (P=0.0501 and P=0.0080, respectively). Patients with diminished expression of E-cadherin as grade II or III had shorter survival period than those with normally expressing and grade I, no significance existed between grade I and grade II or III with respect to different status of E-cadherin expression. Furthermore, Correlation analysis showed level of E-cadherin correlated with that of beta-catenin (P=0.005). Cox proportional hazards model analysis suggested downregulation of E-cadherin was an important factor indicating poor prognosis. CONCLUSION: As a probable independent prognostic factor, it correlates with overall and disease free survival period, expression of E-cadherin but not beta catenin may predict prognosis in patients with ESCC. PMID- 12532438 TI - Expression of 1A6 gene and its correlation with intestinal gastric carcinoma. AB - AIM: To investigate the expression of 1A6 gene in the lesions during the development of intestinal gastric carcinoma. METHODS: One hundred and thirty-six cases of intestinal metaplasia (IM) from surgical resections and biopsy were classified by mucous staining. Expression of 1A6 in all cases was detected using immunohistochemical S-P method. RESULTS: The positive rates of 1A6 in normal and superficial gastritis (SG), severe atrophic gastritis (SAG), type I, II, III IM, dysplasia (Dys) and intestinal gastric carcinoma (IGC) were 12.2 %, 16.7 %, 7.1 %, 22.6 %, 47.8 %, 46.9 % and 60.8 %, respectively. A significant difference among type III IM and SG, SAG, type I and II IM was found (P<0.01), the difference between type III and Dys, IGC being not significant. CONCLUSION: As a new tumor-related gene, expression of 1A6 may be an effective parameter to predict the malignant transformation of precancerous lesion to gastric carcinoma. PMID- 12532437 TI - TK gene combined with mIL-2 and mGM-CSF genes in treatment of gastric cancer. AB - AIM: Cancer gene therapy has received more and more attentions in the recent decade. Various systems of gene therapy for cancer have been developed. One of the most promising choices is the suicide gene. The product of thymidine kinase (TK) gene can convert ganciclovir (GCV) to phosphorylated GCV, which inhibits the synthesis of cell DNA, and then induces the cells to death. Cytokines play an important role in anti-tumor immunity. This experiment was designed to combine the TK gene and mIL-2/mGM-CSF genes to treat gastric cancer, and was expected to produce a marked anti-tumor effect. METHODS: TK gene was constructed into the retroviral vector pLxSN, and the mIL-2 and mGM-CSF genes were inserted into the eukaryotic expressing vector pIRES. The gastric cancer cells were transfected by retroviral serum that was harvested from the package cells. In vitro study, the transfected gastric cancer cells were maintained in the GCV- contained medium, to assay the cell killing effect and bystander effect. In vivo experiment, retroviral serum and cytokines plasmid were transfected into tumor-bearing mice, to observe the changes of tumor volumes and survival of the mice. RESULTS: In vitro experiment, 20 % TK gene transduced cells could cause 70-80 % of total cells to death. In vivo results showed that there was no treatment effect in control group and TK/GCV could inhibit the tumor growth. The strongest anti-tumor effect was shown in TK+mIL-2+mGM-CSF group. The pathologic examination showed necrosis of the cancer in the treated groups. CONCLUSION: TK/GCV can kill tumor cells and inhibit the tumor growth in vivo. IL-2 and GM-CSF strongly enhance the anti-tumor effect. Through the retrovirus and liposome methods, the suicide gene and cytokine genes are all expressed in the tissues. PMID- 12532439 TI - The study of chemiluminescence in gastric and colonic carcinoma cell lines treated by anti-tumor drugs. AB - AIM: To study the influence of chemotherapy on proliferation activation of tumor cell by observing the change of chemiluminescence (CL) and cell cycle in various tumor cell lines after mitomycin C treated. METHODS: BGC823 and LoVo cell lines were all cultured in RPMI-1640, and then were adjusted to a concentration of 1X10(5) cells/ml in fresh media and incubated for 24 h. Mitomycin C (100 ng.ml( 1)) was added to each bottle. All indeses were examined after 24 h. No Mitomycin C was added in control group. Each group contained 8 samples. Flow cytometric analysis and luminol-dependent CL were used to investigate the effect of mitomycin C on two gastrointestinal carcinoma cell lines. RESULTS: BGC823 and LoVo cell lines incubated with MMC for 24 h. We discovered that the emergence of peak of CL stimulated by PHA was postponed significantly (BGC823: 12.63+/-3.21 vs 4.50+/-1.04, LoVo: 13.25+/-2.96 vs 5.12+/-1.36, P<0.01) and the peak intension of CL was reduced significantly (BGC823: 120.25+/-16.61 vs 248.38+/-29.17, LoVo: 98.13+/-10.49 vs 267.50+/-18.56, P<0.01). The PI of cell lines was decreased significantly (BGC823: 51.87+/-4.82 vs 25.44+/-2.26, LoVo: 47.11+/-1.04 vs 24.23+/-0.37, P<0.01) and the apoptotic fractions changed by contraries (BGC823: 26.25+/-5.29 vs 9.83+/-2.51, LoVo: 33.50+/-3.68 vs 9.63+/-1.44, P<0.01). CONCLUSION: CL can be used to measure activation of tumor cells. We discovered that the ground CL intensions of two cell lines were not high but increased rapidly after stimulation of PHA. The CL peak ranged from 4-5 minutes, and then decreased gradually. The results were not reported before. CL of tumor cell has close correlativity with the dynamics of cell cycle and can reflect the feature of oxidation metabolism and proliferation activation of tumor cell. So it can be used to observe the influence of chemotherapy drug on metabolism and proliferation activation of tumor cell and screen out chemotherapy drugs to which tumor cells are sensitive. PMID- 12532440 TI - Association of cyclooxygenase-2 expression with Hp-cagA infection in gastric cancer. AB - AIM: To observe the expression of cyclooxygenase-2 (COX-2) and to investigate the association between COX-2 expression and infection with cytotoxic-associated gene A (cagA) positive strain Helicobacter pylori (Hp) in human gastric cancer, and subsequently to provide fresh ideas for the early prevention of gastric cancer. METHODS: 32 Specimens of gastric cancer and corresponding adjacent normal gastric mucosa were obtained from patients who had undergone surgical operations of gastric cancer. All the samples including 1 case of stomach malignant lymphoma and 31 cases of gastric adenocarcinoma were confirmed by pathology diagnosis. The expression of COX-2 in 32 specimens of gastric cancer and corresponding adjacent normal gastric mucosa was quantitatively determined and analyzed with Flow Cytometry, and the levels of COX-2 protein were compared between specimens with cagA(+) Hp infection and those without cagA(+) Hp infection. The cagA gene in 32 specimens of gastric cancer was detected by polymerase chain reaction (PCR) method. RESULTS: Twenty-seven of 32 (84 %) specimens of gastric cancer showed over-expression of COX-2, compared with the adjacent normal gastric mucosa. cagA(+) gene were detected from 19 specimens of gastric cancer, but not from the other 13 specimens. The levels of COX-2 protein in 19 specimens of gastric cancer with cagA(+) Hp infection (the number of positive cells was 73.82+/-18.2) were significantly higher than those in the 13 specimens without cagA(+) Hp infection (the number of positive cells was 35.92+/-22.1). CONCLUSION: COX-2 is overexpressed in gastric cancer and cagA(+) Hp infection could up-regulate the expression of COX-2 in gastric cancer in human. There may also exist another way or channel to regulate the expression of COX-2 in gastric cancer in addition to cagA(+) Hp infection. Therefore, applying COX-2 selective inhibitors could be an effective and promising way to prevent gastric cancer. PMID- 12532441 TI - Expression of cyclooxygenase-2 and clinicopathologic features in human gastric adenocarcinoma. AB - AIM: To study the expression of cyclooxygenase-2 (COX-2) gene in gastric cancer and the relationship between COX-2 expression and clinicopathologic features of gastric cancer. METHODS: With reference to the expression of beta-actin gene, COX 2 mRNA level was examined in cancerous tissues and adjacent noncancerous mucosa from 33 patients by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). Quantitation of relative band Adj volume counts was performed using molecular Analyst for windows software. The COX-2 index was determined from the band Adj volume counts ratio of COX-2 to constitutively expressed actin. RESULTS: The COX-2 index in gastric carcinoma was significantly higher than that in normal mucosa (0.5966+/-0.2659 vs 0.2979+/-0.171, u=5.4309, P<0.01). Significantly higher expression of COX-2 mRNA was also observed in patients with lymph node involvement than that in those without (0.6775+/-0.2486 vs 0.4105+/ 0.2182, t=2.9341, P<0.01). Furthermore, the staging in the UICC TNM classification significantly correlated with COX-2 overexpression (F=3.656, P<0.05), the COX-2 index in stage III and IV was significantly higher than those in stage I and II (q=3.2728 and q=3.4906, P<0.05). The COX-2 index showed no correlation with patient's age, sex, blood group, tumor location, gross typing, depth of invasion, differentiation, and the greatest tumor dimension (P>0.05). CONCLUSION: Expression of COX-2 mRNA in gastric carcinoma was significantly higher, which may enhance lymphatic metastasis in patients with gastric carcinoma. The staging in the UICC TNM classification was significantly correlated with COX-2 over-expression. COX-2 may contribute to progression of tumor in human gastric adenocarcinoma. PMID- 12532442 TI - Endoscopic ultrasonography in preoperative staging of gastric cancer: determination of tumor invasion depth, nodal involvement and surgical resectability. AB - AIM: Current study was aimed to evaluate the usefulness of EUS in TNM staging of gastric cancer by comparing EUS preoperative staging with pathological findings, and the preliminary exploration of possible reasons for overstaging and understaging phenomenon was especially intended. METHODS: A total of 35 patients with histologically confirmed gastric adenocarcinoma were referred to EUS and staged preoperatively by using the TNM system. The preoperative endosonographic results were compared with the histopathological staging. RESULTS: The overall accuracy of EUS for determination of the T stage was 80.0 %, and for T1, T2, T3, and T4 was 100 %, 71.4 %, 87.5 % and 72.7 %, respectively. For N stage, EUS had the accuracy of 68.6 %, with sensitivity and specificity of 66.7 % and 73.7 %, respectively. Resectability was predicted with sensitivity and specificity of 87.5 % and 100 %, respectively. CONCLUSION: EUS is an accurate staging modality in most cases, with a few exceptions of overstaging and understaging. Patients with gastric cancers can benefit from preoperative EUS staging for establishing individualized therapy. However, EUS criteria to differentiate benign from malignant nodes still need to be further defined by future studies. PMID- 12532443 TI - Vascularity of hepatic VX2 tumors of rabbits: assessment with conventional power Doppler US and contrast enhanced harmonic power Doppler US. AB - AIM: To investigate the characteristics of the vascularity of hepatic metastasis. METHODS: Six New Zealand rabbits, weighing averagely 2.7+/-0.4 kg, were selected and operated to establish hepatic VX2 tumor carrier model. Hepatic VX2 tumors were then imaged with conventional B mode US, second harmonic imaging (SHI), color Doppler flow imaging (CDFI), power Doppler imaging (PDI) and harmonic PDI by a transducer S8 connected to HP-5 500 ultrasound system. A kind of self made echo contrast agent was intravenously injected at a dose of 0.01 mL/kg through ear vein, and then the venous passage was cleaned with sterilized saline. RESULTS: Totally, 6 hypoechoic lesions and 3 hyperechoic lesions were found in the six carrier rabbits with a mean size about 2.1+/-0.4 cm under conventional B mode ultrasound, they were oval or round in shape with a clear outline or a hypoechoic halo at the margin of the lesions. Contrast agent could not change the echogenicity of the lesions under conventional B mode and SHI, however, it could greatly increase the flow sensitivity of the lesions under PDI and harmonic PDI. Nutrient artery of these metastatic lesions might also be well depicted under contrast enhanced PDI and harmonic PDI. CONCLUSION: Our result suggested that contrast enhanced PDI, especially harmonic PDI, was a promised method in the detection of vascularity of hepatic tumor nodules. PMID- 12532444 TI - Potent inhibition of angiogenesis and liver tumor growth by administration of an aerosol containing a transferrin-liposome-endostatin complex. AB - AIM: To obtain an efficient delivery system for transporting endostatin gene to mouse liver tumor xenografts by administration of aerosol. METHODS: Recombinant plasmid pcDNA3.0/endostatin containing human endostatin gene together with signal peptide from alkaline phosphatase were transferred into human umbilical vein endothelial cell (HUVEC) by transferrin(TF)-liposome-endostatin complex. Western blot was used to detect the expression of human endostatin in transfected HUVEC cells and its medium. After the tumor-bearing mice were administrated with TF liposome-endostatin complex, the lung tissue was analyzed by immunohistochemical method for expression of endostatin and the tumors were treated with CD-31 antibody to detect the density of microvessels in tumor tissues. The inhibition of tumor growth was estimated by the weight of tumors from groups treated with different doses of TF-liposome-endostatin complex. DNA fragmentation assay was used to detect the apoptosis of the cells from primary liver tumor. RESULTS: Western blot analysis and immunohistochemical method confirmed the expression of endostatin protein in vitro and in vivo. After the tumor sections were treated with CD-31 antibody, the positive reaction cells appeared brown while the negative cells were colorless. The positively stained area of the TF-liposome endostatin treated group was significantly smaller (P<0.01, 645.8+/-55.2 microm(2)) than that of the control group (1 325.4+/-198.5 microm(2)). The data showed a significant inhibition of angiogenesis. After administration of TF liposome-endostatin, comparing with the control group administrated with TF liposome-pcDNA3.0, liver tumor growth in the mice treated with 50, 250 and 500 mg DNA/kg was inhibited by 36.6 %, 40.8 %, and 72.8 %, respectively (P<0.01). And a typical DNA fragmentation of apoptosis was found in the cells from tumor tissues of the mice treated with TF-liposome-endostatin but none in the control group. CONCLUSION: Endostatin gene could be efficiently transported into the mice with TF-liposome-DNA delivery system by administration of aerosol. TF-liposome mediated endostatin gene therapy strongly inhibited angiogenesis and the growth of mouse xenograft liver tumors. It also could promote the development of apoptosis of tumors without direct influence on tumor cells. PMID- 12532445 TI - Expression of IGF-II in early experimental hepatocellular carcinomas and its significance in early diagnosis. AB - AIM: To investigate the serum level and expression of insulin growth factor II (IGF-II) in liver tissues of rats with early experimental hepatocellular carcinomas (HCC) and its significance in early diagnosis. METHODS: Early experimental hepatocellular carcinomas were induced by diethylnitrosamine (DENA) in 180 male SD rats. Another 20 male SD rats served as control. The IGF-II serum level was measured by ELISA. Immunohistochemistry and electron microscopic immunohistochemistry were used to observe the expression of IGF-II in normal and tumor liver tissues and its ultrastructural location in malignant hepatocytes. The expressions of IGF-II in human hepatoma cell lines HepG2, SMMC7721 and human embryonic liver cell line L-02 were measured by immunocytochemistry. IGF-II mRNA level was studied by in situ hybridization. RESULTS: IGF-II was expressed in the cytoplasm of both sinusoidal cells in paracancerous cirrhotic liver tissue and malignant hepatocytes in early experimental HCC tissues. Gold particles were seen on the rough endoplasmic reticulum and the mitochondrion in malignant hepatocytes. IGF-II was expressed in the human hepatoma cell lines. The mRNA level of IGF-II was higher in rat liver tumor tissue than in normal rat liver tissue. The serum IGF-II level of the early experimental HCC group was 34.67+/ 10.53 ng.ml(-1) and that of the control group was 11.75+/-5.84 ng.ml(-1). The rank sum test was used for statistical analysis. There was a significant difference between the two groups (P<0.01). CONCLUSION: During the induction of early experimental HCC by DENA, IGF-II may promote hepatocytic proliferation via a paracrine mechanism in the pre-cancerous stage. When hepatocytes are transformed into malignant cells, they may secrete IGF-II and promote malignant cell proliferation by an autocrine mechanism. IGF-II may be a possible biological marker in the early diagnosis of HCC. PMID- 12532446 TI - Study on the mechanism of epidermal growth factor-induced proliferation of hepatoma cells. AB - AIM: Many growth factors, such as epidermal growth factor (EGF), are associated with the carcinogenesis. EGF plays its role in the proliferation of hepatoma cells through binding with EGF receptor (EGFR) and a series of signal transduction. But the postreceptor pathway is still not clear. In the present experiment, we studied the effect of tyrosine kinase, protein kinase C, Na(+)/H(+) exchange, calmodulin and voltage-dependent Ca(2+) channel on EGF induced hepatoma cell proliferation. METHODS: Hepatoma cell line SMMC7721 was cultured in RPMI1640 serum-free medium. In order to study the effect of thyrosine kinase, protein kinase C, Na(+)/H(+) exchange, calmodulin and voltage-dependent Ca(2+) channel on human heptoma cell proliferation induced by epidermal growth factor (EGF), DNA synthesis rate of hepatoma cells was measured by the method of (3)H-TdR incorporation. RESULTS: EGF (10(-9) M) stimulated the proliferation of heptoma cells significantly ((3)H-TdR incorporation was 1 880+/-281 cpm/well, P<0.05), and this effect was significantly inhibited by tyrosine kinase inhibitor genistein ((3)H-TdR incorporation was 808+/-209 cpm/well, P<0.001). Calmodulin inhibitor W-7, protein kinase C inhibitor H-7 and Na(+)/H(+) exchange inhibitor amiloride individually had significant inhibiting effect on EGF-induced proliferation of hepatoma cells ((3)H-TdR incorporation was 978+/-87.3 cpm/well, 1 241+/-147 cpm/well, 1 380+/-189 cpm/well, respectively, P<0.001, P<0.01, P<0.05), but they all had no effect on the basal level proliferation of cultured hepatoma cells ((3)H-TdR incorporation was 1 284+/-260 cpm/well, 1 179+/-150 cpm/well, 1 392+/-152 cpm/well, respectively, (3)H-TdR incorporation of the control was 1 353+/-175 cpm/well, P>0.05). Voltage-dependent Ca(2+) channel inhibitor verapamil had no inhibition on EGF-induced proliferation of hepatoma cells ((3)H-TdR incorporation was 1 637+/-133 cpm/well, P>0.05), it also had no effect on the basal level proliferation of cultured hepatoma cells ((3)H-TdR incorporation was 1 196+/-112 cpm/well, P>0.05). CONCLUSION: Our data suggest that tyrosine kinase, Ca(2+)-calmodulin-dependent pathway, protein kinase C and Na(+)/H(+) exchange play a critical role in EGF-induced proliferation of hepatoma cells and that the effect of EGF is independent of voltage-dependent Ca(2+) channel. PMID- 12532447 TI - The analysis of gamma-glutamyl transpeptidase gene in different type liver tissues. AB - AIM: To probe the value of gamma-glutamyl transpeptidase (GGT) messenger RNA in monitoring canceration of liver cells and for early diagnosis of hepatocellular carcinoma (HCC), by researching the types of GGT messenger RNA (GGTmRNA) in liver tissues and peripheral blood of different hepatopathy. METHODS: The three types of GGTmRNA (A, B, C) in liver tissues and peripheral blood from the patients with HCC, noncancerous hepatopathy, hepatic benign tumor, secondary carcinoma of liver, and healthy persons were detected by reverse-transcription polymerase chain reaction (RT-PCR). RESULTS: (1) In normal liver tissues, type A was predominantly found (100.00 %), type B was not found, type C was found occasionally (25.00 %). (2) The distribution of types of GGTmRNA in liver tissues with acute hepatitis, chronic hepatitis, cirrhosis, alcoholic hepatopathy was similar as in normal liver tissues (P>0.05), but type B was found in 3 of 18 patients with chronic hepatitis (16.67 %), and also in 3 of 11 patients with cirrhosis (27.27 %). (3) There was no significant difference of types of GGTmRNA between liver tissues with hepatic benign tumor, secondary carcinoma of liver and normal liver tissues (P>0.05). (4) Type B was predominant in cancerous tissues with HCC (87.5 %), the prevalence of type B in cancerous tissues was significantly higher than that in normal liver tissues (0/12) (P<0.05), but the prevalence of type A in cancerous tissues (46.88 %) was significantly lower than that in normal liver tissues (100.00 %) (P<0.05), and the prevalence of type C (6.25 %) in cancerous was the same as that in normal liver tissues (25.00 %) (P>0.05). In noncancerous tissues of livers with HCC, the main types were type A and type B, the prevalence of type A (85.71 %, 90.48 %) and type C (14.29 %, 9.52 %) in noncancerous tissues of liver with HCC was similar as that in normal liver tissues (A: 100.00 %; C: 25.00 %) (P>0.05), but the prevalence of type B (80.95 %, 76.19 %) in noncancerous tissues of livers with HCC was significantly higher than that in normal liver tissues (0/12) (P<0.05). (5) The prevalence of type B (37.5 %) in peripheral blood with HCC was higher than that in normal person (0/12) (P<0.05). In peripheral blood, type B was found in 4 of 11 cases of HCC with serum AFP negative. CONCLUSION: The shift of types of GGTmRNA from A to B in liver tissues may be closely related to the development of HCC, and the analysis of GGT gene may provide a useful tool for early diagnosis of HCC. PMID- 12532448 TI - Effects of TNP-470 on proliferation and apoptosis in human colon cancer xenografts in nude mice. AB - AIM: To study the effect of TNP-470 on cell growth, proliferation and apoptosis in human colon cancer xenografts in nude mice. METHODS: Human colon cancer xenografts were transplanted into 20 nude mice. Mice were randomly divided into two groups. TNP-470 treated group received TNP-470(30 mg/kg, s.c) every other day and the control group received a sham injection of same volume saline solution. They were sacrificed after 4 weeks and their tumors were processed for histological examination. The expression of proliferating cell nuclear antigen (PCNA) in tumors was detected using immunohistochemical method with image analysis, and apoptosis in tumor cells was measured by TdT-mediated biotinyated dUTP nick end labeling (TUNEL) staining. RESULTS: Comparing with controls, tumor growth was significantly inhibited in TNP-470 treated group, the inhibitory rate being 54.4 %. Expression of PCNA in tumors of TNP-470 treated group (PI 54.32+/ 11.47) was significantly lower than that of control group (PI 88.54+/-12.36), P<0.01. Apoptosis index (AI) of TNP-470 treated group (18.95+/-1.71) was significantly higher than that of control group (7.26+/-1.44), P<0.001, typical morphological change of apoptosis in tumor cells was observed in TNP-470 treated group. CONCLUSION: Besides the anti-angiogenic effects, TNP-470 can inhibit tumor growth by inhibiting the proliferation and inducing apoptosis of tumor cells. PMID- 12532449 TI - Clinical characteristics and diagnosis of patients with hereditary nonpolyposis colorectal cancer. AB - AIM: To study the clinical characteristics of hereditary nonpolyposis colorectal cancer (HNPCC) in the Chinese population and discuss the identification and management of the patients with HNPCC. METHODS: A series of 140 patients with colorectal cancers (CRC) and HNPCC associated tumors from 30 families fulfilling the Amsterdam criteria were analyzed. RESULTS: A total of 118 patients had CRC. Average age at diagnosis of the first CRC was 45.7 years, 56.8 % and 23.4 % of the first CRC were located proximal to the splenic flexure and in the rectum respectively. Twenty-three (19.5 %) had synchronous and metachronous CRC. Twenty seven patients were found to have extracolonic tumors. Gastric carcinoma was the most common tumor type in our series (44.4 %). CONCLUSION: The frequency of HNPCC was 2.6 % in our series of patients. The main features are an excess of early onset with a propensity to involve the proximal colon, and high frequency of multiple foci. Management and surveillance for these patients should be different from sporadic CRC. Contrary to American and European reports, gastric cancer seems more frequent than endometrial cancer in Chinese. It is necessary to formulate a new HNPCC criterion for Chinese patients. PMID- 12532450 TI - Angiography for diagnosis and treatment of colorectal cancer. AB - AIM: To evaluate the role of preoperative angiography in the diagnosis and treatment of colorectal cancer. METHODS: The authors performed selective arterial cannulation by Seldinger's method in 47 patients to locate the primary cancer and to diagnose metastasis to the liver. Each patient was then given intra-arterial regional chemotherapy, and received 5-fluorouracil (5-Fu, 1 000 mg), mitomycin C (MMC, 20 mg), and cisplatinum (CDDP, 80 mg). RESULTS: The location and shape of each tumor were observed, including metastatic tumors in the liver, in 42 of the 47 (89.4 %) patients. The site of the primary tumor was difficult to identify in 5 cases because the patients had a recurrence of cancer. Arterial chemotherapy was performed successfully in all patients. The authors recorded no partial or significant morbidity resulted from angiography. The only incident was bleeding from the artery puncture site in one patient, which was successfully stopped by general medication. CONCLUSION: Preoperative selective arterial angiography can help the diagnosis and locate primary tumors and to detect liver metastasis. At the same time, regional arterial chemotherapy can be an important form of preoperative therapy. PMID- 12532451 TI - HCV replication in PBMC and its influence on interferon therapy. AB - AIM: To study hepatic virus C (HCV) RNA and HCV protein expression in peripheral blood mononuclear cells (PBMCs) of patients with HCV infection, and explore the relationship between the HCV RNA in the PBMCs and response to interferon (IFN) therapy. METHODS: Type-specific primers were designed and RT-nested PCR was used to detect the plus- and minus- strands of HCV RNA in PBMCs of 54 patients with HCV infection; Indirect immunofluorescence assay was applied to identify HCVNS5 protein expression in PBMCs; 6 month-, 3 MU-IFN regiment was administrated to observe the responses to IFN in 35 chronic hepatitis C patients with different HCV RNA status in PBMCs. RESULTS: HCV plus strand RNA was found in 10 of 19 (52.6 %) acute hepatitis C patients and 22 of 35 (62.9 %) chronic hepatitis C patients. HCV minus strand RNA was detected in 14 of 35 (40.0 %) chronic hepatitis C patients, but only one patient (5.3 %) with acute HCV infection was found to be minus HCV RNA positive. Though no HCV NS5 protein expression was found in the examined 10 cases of acute HCV infection, it was positive in 17 of 20 (85.0 %) chronic hepatitis C patients by indirect immunofluorescence assay. There are significant differences of positive rate of the minus-strand and HCVNS5 protein between acute and chronic hepatitis C groups (u=2.07, P<0.05 and u=4.43, P<0.01 respectively). The patients with minus-strand HCV RNA showed a significantly lower 6-month sustained response (SR-6) to IFN compared to those without minus strand HCVRNA in PBMCs (biologically 14.3 % vs 42.8 %, chi(2)=4.12, P<0.05 and virologically 7.1 % vs 23.9 %, chi(2)=4.24, P<0.05). CONCLUSION: HCV is capable of infecting and replicating in PBMCs, and HCVNS5 protein was expressed in PBMCs. The patients with minus strand HCV RNA in PBMCs showed a significantly lower 6 month sustained response to IFN, suggesting that minus-strand HCV RNA in PBMCs may be one of the factors influencing response to IFN therapy. PMID- 12532452 TI - Targeted ribonuclease can inhibit replication of hepatitis B virus. AB - AIM: To study the effect of a novel targeted ribonuclease (TN), the fusion protein of HBVc and human eosinophil-derived neurotoxin (hEDN), on the HBV replication in vitro. METHODS: The gene encoding the targeted ribonuclease was cloned into pcDNA3.1 (-) to form recombinant eukaryotic expression vector p/TN. Control plasmids, including p/hEDN, p/HBVc, and p/TNmut in which a Lys113-Arg mutation was introduced by sequential PCR to eliminate the ribonuclease activity of hEDN, were also constructed. Liposome-mediated transfection of 2.2.15 cells by p/TN, p/TNmut, p/hEDN, p/HBVc, and pcDNA3.1 (-), or mock transfection was performed. After that, RT-PCR was used to verify the transgene expression. Morphology of the transfected cells was observed and MTT assay was performed to detect the cytotoxicity of transgene expression. Concentration of HBsAg in the supernatant of the transfected cells was measured using solid-phase radioimmunoassay. RESULTS: Transgenes were successfully expressed in 2.2.15 cells. No obvious cytotoxic effect of transgene expression on 2.2.15 cells was found. The HBsAg concentration in the p/TN transfected cells was reduced by 58 % compared with that of mock transfected cells. No such an effect was found in all other controls. CONCLUSION: The targeted ribonuclease can inhibit HBV replication in vitro while it has no cytotoxicity on host cells. The targeted ribonuclease may be used as a novel antiviral agent for human HBV infection. PMID- 12532453 TI - Interaction between hepatitis C virus core protein and translin protein--a possible molecular mechanism for hepatocellular carcinoma and lymphoma caused by hepatitis C virus. AB - AIM: To investigate the interaction between hepatitis C virus core protein and translin protein and its role in the pathogenensis of hepatocellular carcinoma and lymphoma. METHODS: With the components of the yeast two hybrid system 3, "bait" plasmids of HCV core the gene was constructed. After proving that hepatitis C virus core protein could be firmly expressed in AH109 yeast strains, yeast two- hybrid screening was performed by mating AH109 with Y187 that transformed with liver cDNA library plasmids-pACT2 and then plated on quadruple dropout (QDO) medium and then assayed for alpha-gal activity. Sequencing analysis of the genes of library plasmids in yeast colonies that could grow on QDO with alpha-gal activity was performed. The interaction between HCV core protein and the protein we obtained from positive colony was further confirmed by repeating yeast two - hybrid analysis and coimmunoprecipitation in vitro. RESULTS: A gene from a positive colony was the gene of translin, a recombination hotspot binding protein. The interaction between HCV core protein and translin protein could be proved not only in yeast, but also in vitro. CONCLUSION: The core protein of HCV can interact with translin protein. This can partly explain the molecular mechanism for hepatocellular carcinoma and lymphoma caused by HCV. PMID- 12532454 TI - A novel hepatitis B virus mutant with A-to-G at nt551 in the surface antigen gene. AB - AIM: Hepatitis B surface antigen (HBsAg) mutant of hepatitis B virus (HBV) is one of the important factors that result in immune escape and cause failure of immunization. In this study we reported and characterized a novel HBV mutant with A-to-G at nt551 and intended to provide theoretical data for prevention of HBV infection in China. METHODS: A methodology comprising polymerase chain reaction (PCR) amplifying, M13 bacteriophage cloning and nucleotide sequencing was used to analyze the sera of the pediatric patient who was hepatitis B (HB) immune failure. Expression plasmids containing the mutant S gene and a wild-type (adr) S gene were constructed respectively and the recombinant HBsAg were expressed in COS-7 cells under the regulation of SV40 early promoter. The recombinant proteins were investigated for their immunological reactivity with different monoclonal antibodies (mAb) against "a" determinant and vaccine-raised human neutralizing antibodies. RESULTS: It was found that there was a new point mutation at nt551 of the HBV (adr) genome from A to G, leading to a substitution of methionine (Met) to valine (Val) at position 133 in the "a" determinant of HBsAg. Compared to the wild-type HBsAg, the binding activity of the mutant HBsAg to mAbs (A6, A11 and S17) and to vaccine-raised human anti-hepatitis B surface antibody (anti-HBs) decreased significantly. CONCLUSION: According to the facts that the patient has been immunized with HB vaccine and that the serum is anti-HBs positive and HBsAg negative, and based on the nucleotide sequence analysis of the mutant HBV S gene and its alteration of antigenicity, the HBV is considered to be a new vaccine induced immune escape mutant different from the known ones. PMID- 12532455 TI - Evaluation of the string test for the detection of Helicobacter pylori. AB - AIM: Helicobacter pylori can be diagnosed by invasive or non-invasive tests but to obtain bacteria for culture and antibiotic susceptibility testing, an upper GI endoscopy is often required. The string test may be a minimally-invasive alternative method of obtaining H. pylori samples. This study evaluates the sensitivity and specificity of the string test in the diagnosis of H. pylori in comparison with endoscopic means of diagnosis. METHODS: This was a prospective open comparative study of patients with dyspepsia with endoscopy-based tests as gold standard (defined as a positive CLO test and antral histology). Fasting patients swallowed the encapsulated-string (Entero-test Hp), which was withdrawn after 1 hour. The gastric juice from the string was plated onto H. pylori selective media for culture. Helicobacter pylori was identified by typical colony morphology, gram stain and biochemical test results. RESULTS: Thirty dyspeptic patients were recruited of whom 21 (70 %) were positive for H. pylori according to the gold standard. The sensitivity, specificity, positive predictive value, and negative predictive value for the string test were 38 %, 100 %, 100 % and 41 % respectively, and for endoscopic biopsies 81 %, 100 %, 100 %, 69 % respectively (P=0.004). Logistic regression showed that only abundant growth density from endoscopic biopsy cultures to be a predictor of a positive string test (P=0.018). CONCLUSION: The string test is an alternative method to endoscopy in obtaining H. pylori but has a low sensitivity compared to endoscopic biopsies. PMID- 12532456 TI - Angiogenesis effect on rat liver after administration of expression vector encoding vascular endothelial growth factor D. AB - AIM: To verify the expressing efficiency and angiogenesis effect after administration of expression vector encoding for vascular endothelial growth factor D in normal and ischemic rat liver. METHODS: Ten female S-D rats were administrated with liver tissue dot injection of naked PCHO/hVEGF-D, 50 microg/dot, three dots for each. The same amount of physiological saline was used as control in the neighboring lobe. Fourteen S-D rats, using inflow occlusion of left lateral lobe, were divided into two groups, seven rats in each group. One was ischemic plasmid group, which received naked plasmid PCHO/hVEGF-D injection of 150 microg. The other received the equal amount of natural saline injection and designed as control. The expressions of hVEGF-D in mRNA and protein levels were identified by in situ hybridization and immunohistochemistry, respectively. Endothelial cells were labeled by the factor VIII immunohistochemistrically. The average number of peri-sinusoidal capillaries of each group was calculated and compared statistically 8 days after injection. RESULTS: A large amount of hVEGF-D in mRNA level was found in both normal and ischemic plasmid groups and but none in their corresponding control groups. The protein of hVEGF was also highly expressed in both normal and ischemic plasmid groups than in the controls. The mean number of capillaries under microscopy (X200) of the plasmid group and control was 10.2+/-2.78 vs 7.1+/-2.02 (P<0.05), and those of ischemic plasmid group and ischemic control were 7.43+/-1.72 vs 4.71+/-1.11 with statistical difference (P<0.05). CONCLUSION: The naked PCHO/hVEGF-D dot injection to normal, ischemic rat liver can produce comparatively high expression of hVEGF in both protein and mRNA levels, and prominently increase the number of new capillaries around hepatic sinuses. Therefore, it could be another ideal choice for the treatment of ischemic liver diseases. PMID- 12532457 TI - Inhibition on the production of collagen type I, III of activated hepatic stellate cells by antisense TIMP-1 recombinant plasmid. AB - AIM: To investigate the inhibition effects on the production of collagen type I, III secreted by activated rat hepatic stellate cells (rHSCs) by antisense tissue inhibitors of metalloproteinase 1 (TIMP-1) recombinant plasmid through elevating interstitial collagenase activity. METHODS: rHSCs were extracted from normal rat liver by pronase and collagenase digestion and purified by centrifugal elutriation, and were cultured on plastic dishes until they were activated to a myofibroblastic phenotype after 7-10 days. RT-Nest-PCR and gene recombinant techniques were used to construct the rat antisense TIMP-1 recombinant plasmids which can express in eucaryotic cells. The recombinant plasmid and the pcDNA3 empty plasmid were transfected in rHSCs by Effectene (QIAGEN) separately. Cells were selected after growing in DMEM containing 400 microg/ml G418 for 2-3 weeks. Expression of exogenous gene was assessed by Northern blot, and expression of TIMP-1 in rHSCs was determined by Northern blot and Western blot. We tested the interstitial collagenase activity with FITC-labled type I collagen as substrate. Ultimately, we quantified the type I, III collagen by Western blot. RESULTS: The exogenous antisense TIMP-1 recombinant plasmid could be expressed in rHSCs well, which could block the expression of TIMP-1 greatly, the ratio of TIMP-1/GAPDH was 0.67, 2.41, and 2.97 separately at mRNA level (P<0.05); the ratio of TIMP-1/beta actin was 0.31, 0.98 and 1.32 separately at protein level (P<0.05); It might elevate active and latent interstitial collagenase activity, the collagenase activity was 0.3049, 0.1411 and 0.1196 respectively. (P<0.05), which led to promotion the degradation of type I, III collagen, the ratio of collagen I/beta actin was 0.63, 1.78 and 1.92 separately (P<0.05); and the ratio of collagen III/beta-actin was 0.59, 1.81 and 1.98 separately (P<0.05). CONCLUSION: These data shows that the antisense TIMP-1 recombinant plasmid has the inhibitory effects on the production of type I, III collagens secreted by activated rHSCs in vitro. It could be a novel method to reverse hepatic fibrosis in the future. PMID- 12532458 TI - Effect of leflunomide on immunological liver injury in mice. AB - AIM: To study the effect of leflunomide on immunological liver injury (ILI) in mice. METHODS: ILI was induced by tail vein injection of 2.5 mg Bacillus Calmette Guerin (BCG), and 10 d later with 10 microg lipopolysaccharide (LPS) in 0.2 mL saline (BCG+LPS). The alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO) level in plasma and molondiadehyde (MDA), glutathione peroxidase (GSHpx) in liver homogenate were assayed by spectroscopy. The serum content of tumor necrosis factors-alpha (TNF-alpha) was determined by ELISA. Interleukin-1 (IL-1), interleukin-2 (IL-2) and Concanavalin A (ConA)-induced splenocyte proliferation response were determined by methods of (3)H-infiltrated cell proliferation. RESULTS: Leflunomide (4, 12, 36 mg.kg(-1)) was found to significantly decrease the serum transaminase (ALT, AST) activity and MDA content in liver homogenate, and improve reduced GSHpx level of liver homogenate. Leflunomide (4, 12, 36 mg.kg(-1)) significantly lowered TNF-alpha and NO level in serum, and IL-1 produced by intraperitoneal macrophages(PMphi). Moreover, the decreased IL-2 production and ConA-induced splenocyte proliferation response were further inhibited. CONCLUSION: These findings suggested that leflunomide had significant protective action on ILI in mice. PMID- 12532459 TI - Direct intrahepatic portacaval shunt: an experimental study. AB - AIM: To determine the feasibility of creating direct intrahepatic portacaval shunt (DIPS) in swine with puncture under sonographic guidance. METHODS: DIPS was created in 10 domestic swine under sonographic guidance. Liver function, blood ammonia level and portosystemic gradient (PSG) were compared before and after the procedure. Patency of shunt was followed by portography every 7 days after DIPS. RESULTS: DIPS was successfully established in all 10 swine without any complications. One day after procedure the alanine aminotransferase (ALT), aspartate aminotransferase (AST), and blood ammonia level (BAL) of swine rose from 5.40+/-0.69, 16.00+/-0.79 and 35.66+/-4.10 to 34.20+/-3.46, 59.70+/-2.22 and 66.94+/-3.44 respectively. (P<0.05). The PSG decreased from 0.59+/-0.20 kPa to 0.24+/-0.11 kPa after DIPS (P<0.05). The shunt of 10 swine was kept patent from 7 28 days (median patency time was 14 days). CONCLUSION: This initial experience demonstrated that creating intrahepatic portacaval shunt from retrohepatic segment of IVC to portal vein with puncture under sonographic guidance in swine is safe and feasible. Further studies are necessary to perform DIPS in cirrhosis patients. PMID- 12532460 TI - Effects of palmatine on potassium and calcium currents in isolated rat hepatocytes. AB - AIM: To study the effects of palmatine, a known inhibitor on delayed rectifier potassium current and L-type calcium current (I(Ca,L)) in guinea pig ventricular myocytes, on the potassium and calcium currents in isolated rat hepatocytes. METHODS: Tight-seal whole-cell patch-clamp techniques were performed to investigate the effects of palmatine on the delayed outward potassium currents (I(K)), inward rectifier potassium current (I(K1)) and Ca(2+) release-activated Ca(2+) current (I(CRAC)) in enzymatically isolated rat hepatocytes. RESULTS: Palmatine 0.3-100 microM reduced I(K) in a concentration-dependent manner with EC(50) of 41.62+/-10.11 microM and n(H), 0.48+/-0.07 (n=8). The effect of the drug was poorly reversible after washout. When the bath solution was changed to tetraethylammonium (TEA) 8 mM, IK was inhibited. Palmatine 10 microM and 100 microM shifted the I-V curves of I(K) downward, and the block of I(K) was voltage independent. Palmatine 0.3-100 microM also inhibited I(CRAC) in a concentration dependent manner. The fitting parameters were as follows: EC(50)=51.19+/-15.18 microM, and n(H)=0.46+/-0.07 (n=8). The peak value of I(CRAC) in the I-V relationship was decreased by palmatine 10 microM and 100 microM. But the reverse potential of I(CRAC) occurred at Voltage=0 mV in all cells. Palmatine 0.3-100 microM failed to have any significant effect on either inward or outward components of I(K1) at any membrane potential examined. CONCLUSION: The inhibitory effects on I(K) and I(CRAC) could be one of the mechanisms that palmatine exerts protective effect on hepatocytes. PMID- 12532461 TI - Localization of TRAIL/TRAILR in fetal pancreas. AB - AIM: To observe the localization of TRAIL/TRAIR (DR4, DR5, DcR1, DcR2) in the fetal pancreas. METHODS: Fetal pancreas of 32 weeks of pregnancy were obtained from induced abortions, embedded in paraffin, and 4-microm sections were prepared. The localization of TRAIL/TRAILR in fetal pancreas was investigated by fluorescence immunohistochemical method combined with laser scanning confocal microscopy. RESULTS: TRAIL immunoreactive cells were mainly located on the periphery of the pancreas islets. There were a few DcR1 and DcR2 positive cells whereas there were no immunoreactive cells of DR4 and DR5 in the pancreas islets. In the acini and the ducts of the exocrine pancreas there were no TRAIL/TRAILR immunoreactive cells. CONCLUSION: This study not only describes the distribution of TRAIL/TRAILR in the fetal pancreas, but also provides a morphological basis for deducing the function of TRAIL/TRAILR in pancreas, suggesting that in normal pancreatic islets, the pancreatic cells are resistant towards apoptosis too. PMID- 12532462 TI - Influence of platelet activating factor on expression of adhesion molecules in experimental pancreatitis. AB - AIM: To determine whether Platelet activating factor (PAF) has a regulation role in the expression of adhesion molecules and accumulation of neutrophils in a murine model of acute pancreatitis. METHODS: One hundred twenty-eight Kunming mice were divided into four groups. Group 1 received 0.1 ml saline s.c. every hour for three hours (sham). Group 2 received cerulein (50 microg/kg dose s.c.) every hour for three hours. Group 3 received AP and additional challenge of PAF (50 mg/kg in absolute ethanol) (AP/PAF). Group 4 received AP, plus therapeutic treatment with GAB (25 mg dose i.p.) immediately after the first challenge of cerulein (AP/GAB). Animals were sacrificed at 12 h after the first challenge of saline or cerulein. Adhesion molecules of pancreas were semi-quantified by SP methods. Standard assays were performed for serum amylase and myeloperoxidase activity (MPO) of pancreas. Histology of pancreas was scored in a blind manner. Water content of pancreas was also measured at the same time. RESULTS: Control pancreata showed negligible adhesion molecule expression and neutrophil accumulation. There were evident adhesion molecules expression and neutrophil accumulation in AP and AP/PAF compared with sham (P<0.05). AP/GAB had a lower level of adhesion molecules, neutrophils, and water content versus AP and AP/PAF (P<0.05). Histology showed a trend toward improvement in AP/GAB, but did not reach statistical significance. CONCLUSION: PAF can induce the expression of adhesion molecules that mediate neutrophil accumulation. The PAF antagonist reduces the expression of adhesion molecules and the severity of inflammation when given immediately after the induction of mild AP in mice. These results suggest that PAF antagonism may be useful in the treatment of mild pancreatitis after its clinical onset. PMID- 12532463 TI - Co-expression of five genes in E coli for L-phenylalanine in Brevibacterium flavum. AB - AIM: To study the effect of co-expression of ppsA, pckA, aroG, pheA and tyrB genes on the production of L-phenylalanine, and to construct a genetic engineering strain for L-phenylalanine. METHODS: ppsA and pckA genes were amplified from genomic DNA of E. coli by polymerase chain reaction, and then introduced into shuttle vectors between E coli and Brevibacterium flavum to generate constructs pJN2 and pJN5. pJN2 was generated by inserting ppsA and pckA genes into vector pCZ; whereas pJN5 was obtained by introducing ppsA and pckA genes into pCZ-GAB, which was originally constructed for co-expression of aroG, pheA and tyrB genes. The recombinant plasmids were then introduced into B. flavum by electroporation and the transformants were used for L-phenylalanine fermentation. RESULTS: Compared with the original B. flavum cells, all the transformants were showed to have increased five enzyme activities specifically, and have enhanced L-phenylalanine biosynthesis ability variably. pJN5 transformant was observed to have the highest elevation of L-phenylalanine production by a 3.4-fold. Co-expression of ppsA and pckA increased activity of DAHP synthetase significantly. CONCLUSION: Co-expression of ppsA and pckA genes in B. flavum could remarkably increase the expression of DAHP synthetase; Co expression of ppsA, pckA, aroG, pheA and tyrB of E. coli in B. flavum was a feasible approach to construct a strain for phenylalanine production. PMID- 12532464 TI - Tri-iodothyronine supplement protects gut barrier in septic rats. AB - AIM: To investigate the role of tri-iodothyronine supplement in protecting gut barrier in septic rats. METHODS: Twenty-two rats were randomized into three groups: sham group (n=6), sepsis group (n=8), and sepsis plus tri-iodothyronine (T(3)) group (n=8). Septic rat model was established through cecal ligation and puncture (CLP). After 5 h, sham and sepsis groups received saline, and the remaining group received T(3) intraperitoneally. Twenty-one hrs After CLP, intestinal permeability and serum free T(3) and T(4) were measured with fluorescence spectrophotometer and by radioimmunoassay, respectively. Intestinal ultrastructure and histologic morphology were observed under transmission electron microscopy (TEM) and light microscopy, respectively. RESULTS: After 21 h, septic symptoms and signs in sepsis plus T(3) group were milder than those in sepsis group. Serum FT(3) or FT(4) concentration in sepsis group was lower than that in sham group (1.59+/-0.20, 3.41+/-2.14 pmol/L vs 3.44+/-1.40, 9.53+/-3.39 pmol/L, P<0.05), and FT(3) concentration in sepsis plus T(3) group (3.40+/-1.65 pmol/L, P<0.05) was corrected. Portal concentration of fluorescein isothiocyanate dextran (FITC-D) in sepsis group (2.51+/-0.56 mg/L) was higher than that in sham group (1.22+/-0.21 mg/L) (P<0.01), and in sepsis plus T(3) group (1.68+/-0.38 mg/L) it was decreased significantly(P<0.01). TEM and light microscopy showed that T(3) supplement preserved well ultrastructure and morphology of intestinal mucosa in septic rats. CONCLUSION: Tri-iodothyronine supplement protects gut barrier in septic rats. PMID- 12532465 TI - Effects of cholesterol on the phenotype of rabbit bile duct fibroblasts. AB - AIM: To investigate how cholesterol (Ch) can affect the phenotype of bile duct fibroblasts of New Zealand rabbits. METHODS: 16 rabbits were divided randomly into two groups: the control group and the experiment group. The rabbits in experiment group were fed with hypercholesterol diet for 8 weeks. Bile duct was dissociated from rabbits and prepared for transmission electron microscopy. The purified bile duct fibroblasts were cultured and divided randomly into three groups: control group, Ch middle concentration group (0.6 g/L), Ch high concentration group (1.2 g/L). After incubated for 72 h, the fibroblasts were made into specimens for transmission electron microscopy. The expression of alpha actin in bile duct fibroblasts was measured by means of laser scanning confocal microscopy. RESULTS: With the transmission electron microscopy, the normal bile duct fibroblasts were shuttle-shaped, and there were abundant rough endoplasmic reticulums (RER), but few mitochondria or microfilaments in cytoplasm. This is the typical phenotype of fibroblasts. Bile duct fibroblasts of hypercholesterolemic rabbits were observed. by the transmission electron microscopy Rough endoplasmic reticulums were significantly reduced, with a lot of microfilament bundles or stress fibers appeared in cytoplasm, especially under plasma membrane. Dense bodies were scattered within these bundles. Macula densas and discontinuous sarcolemma were found under plasma membrane. It suggested that the bile duct fibroblasts of hypercholesterolemic rabbits presented the phenotype of smooth muscle cell. The cultured bile duct fibroblasts also had typical phenotype of fibroblasts. After stimulated by middle concentration cholesterol (0.6 g/L) for 72 h, there appeared lots of microfilaments in cytoplasm, but without dense body, macula densa and discontinuous sarcolemma. Observed with confocal microscopy, there were many regular bundles of microfilaments in fibroblasts treated with middle concentration ch (0.6 g/L) and the expression of alpha-actin was significantly increased. The average fluorescence value of middle concentration group was 1 628+/-189 (P<0.01 vs control group). Microfilaments and the expression of alpha-actin were greatly decreased in fibroblasts of high concentration group (1.2 g/L). The average fluorescence value of high concentration group was 1 427+/-153 (P<0.05 vs middle concentration group). There were a lower expression of alpha-actin and few microfilaments in bile duct fibroblasts of control group with an average fluorescence value of 1 224+/-138. CONCLUSION: Cholesterol can make bile duct fibroblasts have the phenotypic characteristics of smooth muscle cell both in vitro and in vivo and this effect is more significant in vivo. The effect is probably associated with some other factors besides cholesterol. PMID- 12532466 TI - The effect of famotidine on gastroesophageal and duodeno-gastro-esophageal refluxes in critically ill patients. AB - AIM: To investigate the effect of famotidine on gastroesophageal reflux (GER) and duodeno-gastro-esophageal reflux (DGER) and to explore it's possible mechanisms. To identify the relevant factors of the reflux. METHODS: Nineteen critically ill patients were consecutively enrolled in the study. Dynamic 24 hours monitoring of GER and DGER before and after administration of famotidine was performed. The parameters of gastric residual volume, multiple organ disorder syndrome (MODS) score, acute physiology and chronic health evaluation II (APACHE II) score and PEEP were recorded. Paired t test; Wilcoxon signed ranks test and Univariate analysis with Spearman's rank correlation were applied to analyse the data. RESULTS: Statistical significance of longest acid reflux, reflux time of pH<4 and fraction time of acid reflux was observed in ten critically ill patients before and after administration. P value is 0.037, 0.005, 0.005 respectively. Significance change of all bile reflux parameters was observed before and after administration. P value is 0.007,0.024, 0.005, 0.007, 0.005. GER has positive correlation with APACHE II score and gastric residual volume with correlation coefficient of 0.720, 0.932 respectively. CONCLUSION: GER and DGER are much improved after the administration of famotidine. GER is correlated with APACHE II score and gastric residual volume. PMID- 12532467 TI - Different alterations of cytochrome P450 3A4 isoform and its gene expression in livers of patients with chronic liver diseases. AB - AIM: To determine whether parenchymal cells or hepatic cytochrome P450 protein was changed in chronic liver diseases, and to compare the difference of CYP3A4 enzyme and its gene expression between patients with hepatic cirrhosis and obstructive jaundice, and to investigate the pharmacologic significance behind this difference. METHODS: Liver samples were obtained from patients undergoing hepatic surgery with hepatic cirrhosis (n=6) and obstructive jaundice (n=6) and hepatic angeioma (controls, n=6). CYP3A4 activity and protein were determined by Nash and western blotting using specific polychonal antibody, respectively. Total hepatic RNA was extracted and CYP3A4cDNA probe was prepared according the method of random primer marking, and difference of cyp3a4 expression was compared among those patients by Northern blotting. RESULTS: Compared to control group, the CYP3A4 activity and protein in liver tissue among patients with cirrhosis were evidently reduced. (P<0.01) Northern blot showed the same change in its mRNA levels. In contrast, the isoenzyme and its gene expression were not changed among patients with obstructive jaundice. CONCLUSION: Hepatic levels of P450s and its CYP3A4 isoform activity were selectively changed in different chronic liver diseases. CYP3A4 isoenzyme and its activity declined among patients with hepatic cirrhosis as expression of cyp3a4 gene was significantly reduced. Liver's ability to eliminate many clinical therapeutic drug substrates would decline consequently. These findings may have practical implications for the use of drugs in patients with cirrhosis and emphasize the need to understand the metabolic fate of therapeutic compounds. Elucidation of the reasons for these different changes in hepatic CYP3A4 may provide insight into more fundamental aspects and mechanisms of imparied liver function. PMID- 12532468 TI - Changes in the level of serum liver enzymes after laparoscopic surgery. AB - AIM: The purpose of this study was to investigate the effect of laparoscopic surgery on liver function in humans and the possible mechanisms behind such effect. METHODS: Blood samples from 286 patients who underwent laparoscopic cholecystectomy (LC) and 40 patients who underwent open cholecystectomy (OC) were tested for liver function by measuring the level of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) before and after the operations. The same tests were also applied to 18 laparoscopic colorectal cancer resection (LCR) patients and 23 open colorectal cancer resection (OCR) patients to determine whether CO(2) pneumoperitoneum could alter the serum liver enzymes. RESULTS: The level of serum ALT and AST increased significantly during the first 48 hours post operations in both LC and LCR patients. However, no significant change of the serum liver enzymes was detected in both OC and OCR patients. As a result, there was statistically significant difference in change of both ALT and AST levels between LC and OC patients and LCR and OCR patients, respectively. By the 7(th) day post operation, the level of both enzymes returned to normal values in LC, OC and OCR patients except LCR patients whose enzymes remained at a higher level. CONCLUSION: Transient elevation of hepatic transaminases occurred after laparoscopic surgery. The major causative factor seemed to be the CO(2) pneumoperitoneum. In most of the laparoscopic surgery patients, the transient elevation of serum liver enzymes showed no apparent clinical implications. However, if preoperative liver function was very poor, laparoscopic surgery may not be the best choice for the treatment of patients with certain abdominal diseases. PMID- 12532470 TI - Effects of time interval for hemofiltration on the prognosis of severe acute pancreatitis. AB - AIM: To evaluate the impact of time interval for hemofiltration (HF) on the prognosis of severe acute pancreatitis (SAP). METHODS: Thirty-six consecutive patients with severe acute pancreatitis were included in the study. Atlanta classification system was applied for stratification. They were randomly divided into short veno-venous HF group, (SVVH, Group 1, 20 patients); and long veno venous HF group (LVVH, Group 2, 16 patients). In group 1, SVVH was stopped when the abdominal signs disappeared, and heart rate and breath rate were less than 90 beats/min and 20 times/min, respectively. HF was stopped if SVVH was continued, and when heart rate and breath rate were more than 90 beats/min and 20 times/min again (Group 2). Except that the time interval for HF was different, other parameters for HF were the same. And conservative curing rate, survival rate, cost for hospital stay and length of hospital stay were observed. RESULTS: Time interval for HF in Group 1 (3.81+/-1.3 hr) was shorter than that of in Group 2 (9.38+/-2.9 hr), P<0.01. Conservative curing rate (90 %) in Group 1 was much higher than that in Group 2 (56.3 %) (P<0.05); but cost in Group 1 (RMB 56 600+/ 56 400 Yuan) was lower than that in Group 2 (RMB 137 000+/-105 000 Yuan) (P<0.05). And the survival rate (95 %) in Group 1 was higher than that in Group 2 (81.3 %) (P<0.25); however, hospital stay in Group 1 (44.3+/-41 days) was shorter than that in Group 2 (55.2+/-39.5 days) (P<0.2). So, the prognosis was not improved through the prolongation of time interval for HF, but side-effects were seen. CONCLUSION: The prognosis was not further improved by LVVH in the treatment of SAP, with side-effects. Time interval for HF plays an important role in treatment of SAP in early stage. SVVH is thought to be superior to LVVH; and LVVH is superior to CVVH in early (72 hrs) treatment of SAP. PMID- 12532469 TI - Relationship between expression of E-cadherin-catenin complex and clinicopathologic characteristics of pancreatic cancer. AB - AIM: To investigate the expression of E-cadherin and alpha-catenin and beta catenin in pancreatic carcinoma and its relationship with the clinicopathologic characteristics, and clarify the mechanism of invasion and metastasis of pancreatic cancer. METHODS: The expression of E-cadherin and alpha-, beta-catenin was examined in 47 cases of infiltrative ductal adenocarcinoma of pancreas and 12 adult normal pancreatic tissues by immunohistochemical technique. RESULTS: The immunoreactivity of E-cadherin and alpha-, beta-catenin was expressed by normal ductal and acinar cells with strong membranous staining at the intercellular border in 12 cases of adult normal pancreatic tissues. Abnormal expression of E cadherin and alpha-, beta-catenin in 47 pancreatic carcinoma tissues was demonstrated in 53.2 %, 61.7 % and 68.1 %, respectively. Both abnormal expression of E-cadherin and alpha-catenin significantly correlated with differentiation, lymph node and liver metastases (P<0.05, respectively), whereas aberrant beta catenin expression only correlated with lymph node and liver metastases (P<0.001). Abnormal E-cadherin and alpha-, beta-catenin expression was not associated with tumor size, invasion and survival time of patients (P>0.05, all). CONCLUSION: Pancreatic cancer likely occurs in case of E-cadherin-catenin complex genes mutations or deletions and abnormal expression of proteins, which significantly correlate with the biologic character of the tumor and lymph node and liver metastases. It is suggested that the abnormal E-cadherin-catenin complex expression plays an important role in the development and progression of tumor, and thus may become a new marker in pancreatic cancer metastasis. PMID- 12532471 TI - Expression of p57kip2, Rb protein and PCNA and their relationships with clinicopathology in human pancreatic cancer. AB - AIM: To investigate the effects of inhibiting factor of cell cycle regulation p57(kip2), retinoblastinoma protein (Rb protein) and proliferating cell nuclear antigen (PCNA) in the genesis and progression of human pancreatic cancer. METHODS: The expression of p57(kip2), Rb protein and PCNA in tumor tissues and adjacent tissues of 32 patients with pancreatic cancer was detected with SP immunohistochemical technique. RESULTS: p57(kip2) protein positive-expression rate in tumor tissues of pancreatic cancer was 46.9 %, which was lower than that in adjacent pancreatic tissues (75.0 %) (chi(2)=5.317, P<0.05), p57(kip2) protein positive-expression correlated significantly with tumor cell differentiation (well-differentiation versus moderate or low-differentiation, P<0.05) but did not correlate significantly with lymph node metastasis (lymph node metastasis versus non-lymph node metastasis, P>0.05); Rb gene protein positive-expression rate in tumor tissues was 50.0 %, which was also lower than that in adjacent pancreatic tissues (78.1 %) (chi(2)=5.497, P<0.05); PCNA positive-expression rate was 71.9 %, being higher than that in adjacent pancreatic tissues (43.8 %) (chi(2)=5.189, P<0.05), PCNA positive-expression also correlated significantly with tumor cell differentiation and lymph node metastasis (well-differentiation versus moderate or low- differentiation, lymph node metastasis versus non-lymph node metastasis, P<0.05). Rb protein positive-expression rate in the tumor tissues of p57(kip2) protein positive-expression group was 53.3 %; and Rb protein positive-expression rate in the tumor tissues of p57(kip2) protein negative-expression group was 47.1 %. There was no significant relationship between the two groups (r=0.16507, P>0.05). CONCLUSION: The decreased expression of p57(kip2), Rb protein or over expression of PCNA protein might contribute to the genesis or progression of pancreatic cancer, p57(kip2), Rb protein and PCNA may play an important role in genesis and progression of pancreatic cancer. PMID- 12532472 TI - One-stage urethral reconstruction using colonic mucosa graft: an experimental and clinical study. AB - AIM: To investigate the possibility of urethral reconstruction with a free colonic mucosa graft and to present our preliminary experience with urethral substitution using a free graft of colonic mucosa for treatment of 7 patients with complex urethral stricture of a long segment. METHODS: Ten female dogs underwent a procedure in which the urethral mucosa was totally removed and replaced with a free graft of colonic mucosa. A urodynamic study was performed before the operation and sacrifice. The dogs were sacrificed 8 to 16 weeks after the operation for histological examination of urethra. Besides, 7 patients with complex urethral stricture of a long segment were treated by urethroplasty with the use of a colonic mucosal graft. The cases had undergone an average of 3 previous unsuccessful repairs. Urethral reconstruction with a free graft of colonic mucosa ranged from 10 to 17 cm (mean 13.1 cm). Follow-up included urethrography, urethroscopy and uroflowmetry. RESULTS: Urethral stricture developed in 1 dog. The results of urodynamic studies showed that the difference in the maximum urethral pressure between the pre-operation and pre-sacrifice in the remaining 9 dogs was not of significance (P>0.05). Histological examination revealed that the colonic free mucosa survived inside the urethral lumen of the 10 experimental dogs. Plicae surface and unilaminar cylindric epithelium of the colonic mucosa was observed in dogs sacrificed 8 weeks after the operation. The plicae surface and unilaminar cylindric epithelium of the colonic mucosa was not observed, and metaplastic transitional epithelium covered a large proportion of the urethral mucosa in dogs sacrificed 12 weeks after the operation. Clinically, the patients were followed up for 3-18 months postoperatively (mean 8.5 months). Meatal stenosis was developed in 1 patient 3 months postoperatively and needed reoperation. The patient was voiding very well with urinary peak flow 28.7 ml/s during the follow-up of 9 months after reoperation. The other patients were voiding well with urinary peak flow greater than 15 ml/s. Urethrogram revealed a patent urethra with an adequate lumen with no significant graft sacculation. Neither necrosis of neourethral mucosa nor stenosis at the anastomosis sites has been observed on urethroscopy in 4 patients over 6 months after operation. CONCLUSION: Urethral mucosa can be replaced by colonic mucosa without damaging the continence mechanism in female dogs. Colonic mucosa graft urethral substitution is a feasible procedure for the treatment of complex urethral stricture of a long segment. The technique may be considered when more conventional options have failed or are contraindicated. PMID- 12532473 TI - [Prospects and problems of using geoinformation systems in support of medical supply to troops]. PMID- 12532474 TI - [Resource-saving technologies in increasing the effectiveness of using the hospital bed fund]. AB - To substantiate the resource-saving technologies of more effective use of the hospital fund acceptable for the military health service the expert method was used. During the investigation the chiefs of military hospitals and their deputies in the field of medical work who in 1998-2000 studied in the State Institute for Physicians' Advanced Training filled in the special cards of expert evaluation. The investigation has shown the following main directions of more effective use of the hospital bed fund in the stationary treatment-and prophylactic institutions: the integration of polyclinic and hospital links; the development of hospital-substituting technologies; the use of internal reserves in the stationary treatment-and-prophylactic institutions; the organization of work of the stationary treatment-and-prophylactic institutions of the different structures and departments in a single system of logistical support. PMID- 12532475 TI - [Increase in surgical safety]. AB - The authors discuss the problem of operation period safety. It is postulated that the patients' safety depends on the members of operative team. To analyze this aspect the method of theory of probabilities was used. The significant dependence of complication probability on the duration of operation period and associated factor of tiredness of surgical staff is shown in the article. The way of forming the operative team taking into account the psychophysical qualities of its members is proposed. PMID- 12532476 TI - [Local treatment of wounds by transdermal means based on netted reticulum]. PMID- 12532477 TI - [Complex treatment of trophic ulcers in a day surgical hospital]. PMID- 12532478 TI - [Urgent problems of medical aid in infectious endocarditis]. AB - The authors analyze the incidence of infectious endocarditis (IE), the influence of diagnostic time on the lethality, the significance of surgical treatment. The main ways of further improvement of IE patient therapy are defined. They include the training of medical staff, centralization of the patients in the military medical institutions of the center by transfer from the districts after making the diagnosis in order to conduct the adequate medicamentous and appropriate surgical treatment. PMID- 12532479 TI - [Computerized retinotomography--a method for objective documentation of the efficacy of early diagnosis and treatment of primary open-angle glaucoma]. PMID- 12532480 TI - [Laparoscopic radical prostatectomy]. PMID- 12532481 TI - [Detection of localized prostate cancer during multifocal biopsy]. PMID- 12532482 TI - [Experience in treating patients with myelodysplastic syndromes with cyclosporin A]. PMID- 12532483 TI - [Effectiveness of rispolept in mental disorders in patients with Alzheimer and vascular types of dementia]. PMID- 12532484 TI - [Ecology and clinico-epidemiologic characteristics of intestinal anthroponosis in troops]. AB - The change of the dominant strains of intestinal viruses in the different regions of Russia during 1980-2001 was analyzed. In the North-Western regions the following dynamics was observed: enteroviruses ECHO 4, 6, 14, 30, Koksaki B 5, 6, ECHO 4, 11, 20, rotaviruses, new enteroviruses (serotypes 68-71). The last years the polytype of agents is registered. The prevalence of enteroviruses Koksaki A5, B5, ECHO 7, 13, 24, 30, new enteroviruses (serotype 70) is observed in the Far Eastern region of Russia. Koksaki A, B, ECHO, new enteroviruses (serotypes 70, 71) were noted in the Central and Southern regions. The standard bacteriological examination of 158 patients who had come from the epidemic focus of acute intestinal infections (AII) showed that the agents were in 33 (21%) patients (in 19%--Sh. Flexneri 2a and in 2%--EPIR O124). In the remaining 125 (79%) persons the etiological agent wasn't detected. The additional investigation of fecal and blood samples obtained from all admitted patients showed that among the AII agents of uncertain etiology rotaviruses (49%), ECHO viruses (18%) and Koksaki A (8%) prevailed. PMID- 12532485 TI - [Basic positions of the Federal law "State forensic expert activity in the Russian Federation"]. PMID- 12532486 TI - [Military-medical service of the Federal agency of government contacts and information at the President of the Russian Federation--10 years]. PMID- 12532487 TI - [On the 60th anniversary of the Military Medical museum of the Russian Federation's Ministry of Defense]. PMID- 12532488 TI - [Role of the Northern fleet medical laboratory in solving urgent problems of medical support]. PMID- 12532489 TI - LASIK--the Indian eye controversy. PMID- 12532490 TI - Gene therapy in medicine. PMID- 12532491 TI - Complications of laser-in-situ-keratomileusis. AB - Laser-in-situ-keratomileusis (LASIK) has become a popular technique of refractive surgery because of lower postoperative discomfort, early visual rehabilitation and decreased postoperative haze. Compared to photorefractive keratectomy (PRK), LASIK involves an additional procedure of creating a corneal flap. This may result in complications related to the flap, interface and underlying stromal bed. The common flap-related complications include thin flap, button holing, free caps, flap dislocation and flap striae. The interface complications of diffuse lamellar keratitis, epithelial ingrowth and microbial keratitis are potentially sight threatening. Compared to PRK, there is less inflammation and faster healing after LASIK, but there is a longer period of sensory denervation leading to the complication of dry eyes. The refractive complications include undercorrection, regression, irregular astigmatism, decentration and visual aberrations. Honest and unbiased reporting is important to understand the aetiology and redefine the management. PMID- 12532492 TI - Ophthalmic complications and management of Stevens-Johnson syndrome at a tertiary eye care centre in south India. AB - PURPOSE: To review the possible aetiological factors, ocular complications and their management in patients of Stevens-Johnson syndrome with ocular involvement, seen at a tertiary eye care centre. METHODS: We retrospectively reviewed the medical records of patients with Stevens-Johnson syndrome seen between 1987-1998 at L V Prasad Eye Institute. The demographic and possible aetiological factors data causing Stevens-Johnson syndrome were collected. The details of the ocular examination and treatment were collected and examined to determine the pattern of presentation, complications, treatment response and outcome. RESULTS: A total of 95 patients, 40 males (42.10%) and 55 females (57.89%), were identified during the 11-year period. A majority of the patients (n = 53; 55.78%) were between 20 and 40 years of age. All patients had bilateral involvement and most (n = 93; 97.89%) had bilateral symmetrical presentation. The duration from the onset of symptoms to the time of presentation at the institute varied from 6 days to 18 years with most patients presenting after one year (n = 39; 41.05%). The most commonly identified possible causative factor was drugs (n = 55; 51.89%). No definitive cause was identified in 37 (38.94%) patients, and 3 (3.15%) patients had a history of viral fever preceding the onset of Stevens-Johnson syndrome. The best corrected visual acuity at initial presentation was 6/12 or better in 32 (33.68%) patients. Lid abnormalities were observed in 87 (91.51%) patients, conjunctival abnormalities in 92 (96.84%) and corneal complications in 93 (97.89%). All patients were managed medically and 26 (27.36%) patients underwent surgery. CONCLUSION: Stevens-Johnson syndrome remains an important cause of severe visual loss and ocular morbidity, both of which significantly affect the quality of life. Not many medical or surgical options are available even in tertiary eye-care centres. Future advances in immune modulation techniques may prevent many of the sequelae that continue to occur despite the best possible medical care. PMID- 12532493 TI - Mitomycin-C-augmented trabeculectomy for neovascular glaucoma. A preliminary report. AB - PURPOSE: This study aimed to investigate the safety and efficacy of trabeculectomy with intraoperative mitomycin C (MMC) in the management of eyes with neovascular glaucoma (NVG). METHODS: Fifteen eyes of 14 patients with NVG were included in the study. NVG was secondary to central retinal vein occlusion (3 eyes), hemiretinal vein occlusion (2 eyes), proliferative diabetic retinopathy (8 eyes), branch retinal vein occlusion (1 eye) and idiopathic (1 eye). Preoperative retinal ablation was performed in eyes with evidence of posterior segment ischaemia. Following this, all eyes underwent trabeculectomy with intraoperative MMC (0.4 mg/ml for 3 minutes). Clinical outcome assessment included visual acuity, intraocular pressure (IOP), bleb appearance, identification of complications and antiglaucoma medications required to control IOP. RESULTS: The mean IOP decreased from 38.6 +/- 12.9 mmHg (range, 15-64 mmHg) to 17.4 +/- 9.33 mmHg (range, 4-34 mmHg) (P = 0.001). Preoperative visual acuity ranged from light perception to 6/9 in the affected eye. Thirteen (86.6%) of 15 eyes improved vision or retained preoperative vision, one (6.7%) eye lost light perception and one (6.7%) eye developed tractional retinal detachment two years after trabeculectomy. Ten (66.7%) of 15 eyes were classified as surgical success with a mean follow-up of 28.6 +/- 26.3 months (range, 2-82 months). None of the patients developed choroidal haemorrhage, hypotony maculopathy, late onset bleb leak or endophthalmitis. CONCLUSION: Trabeculectomy with intraoperative MMC is a good treatment modality in the management of eyes with NVG. PMID- 12532494 TI - Microbiological study of neonatal conjunctivitis with special reference to Chlamydia trachomatis. AB - PURPOSE: To study the microbial agents, chiefly Chlamydia trachomatis and other bacteria, in neonatal conjunctivitis. METHODS: Conjunctival specimens from 70 newborns with conjunctivitis were subjected to bacterial culture and sensitivity testing, monoclonal antibody based C. trachomatis antigen detection test and species-specific Chlamydia antibody detection in the sera of babies and their mothers, by micro-immunofluorescence assay. RESULTS: Bacteria were isolated from 35 (50%) babies; the majority (20, 57.14%) were Staphylococcus epidermidis. C. trachomatis antigen was detected in conjunctival smears of 17 (24%) babies, and 6 (35.29%) of them were positive for other bacteria. Six babies and their mothers tested positive for C. trachomatis Ig G antibodies. At follow-up after 14 weeks, 6 (35.29%) of the Chlamydia antigen-positive babies were found to have developed recurrent conjunctivitis. CONCLUSION: C. trachomatis is responsible for almost a quarter of all cases of neonatal conjunctivitis, with recurrences in 35% of cases. Bacteria could be isolated from 50% of the patients though the exact role of Staphylococcus epidermidis, isolated from 28.65% of the neonatal conjunctivitis cases, remains unclear. PMID- 12532495 TI - Conjunctival impression cytology in contact lens wearers. AB - PURPOSE: To evaluate the cytological changes in conjunctiva following regular contact lens wear and to determine the correlation, if any, between severity of cytological alteration and symptoms related to contact lens wear. METHODS: One hundred eyes (50 normal asymptomatic subjects) who served as a control group were studied by conjunctival impression cytology (CIC). These subjects were fitted with rigid gas permeable (RGP) or soft contact lenses and were followed up at the end of 3 and 6 months. At each follow-up visit the CIC was repeated. A filter paper with the impression specimen was stained with periodic acid schiff (PAS) and haematoxylin stain to study goblet cell loss. Papanicolaou stain was done to study squamous metaplasia. The cytological changes were graded using the system described by Natadisastra et al. RESULTS: Severity of cytological changes increased with the duration of contact lens wear (P = 0.00001). At the end of 6 months, 60% of symptomatic eyes wearing soft contact lens and RGP lens showed abnormal CIC changes. None of the asymptomatic RGP contact lens wearing eyes showed abnormal CIC changes whereas 33.4% of the asymptomatic soft contact lens wearing eyes showed abnormal CIC changes (P = 0.033). Epithelial changes occurred within 3-6 months of contact lens fitting. CONCLUSION: Severity of cytological changes increased with duration of lens wear (P = 0.00001). Prevalence and severity of cytological alteration is more in symptomatic contact lens wearers. Soft contact lens wearers although asymptomatic showed severe CIC changes. PMID- 12532496 TI - Ocular manifestations of congenital rubella syndrome in a developing country. AB - PURPOSE: To describe the ocular manifestations of congenital rubella syndrome (CRS), a common cause of congenital cataracts in developing countries. METHODS: Retrospective analysis of case records of 46 sero-positive infants under 12 months of age who presented at Aravind Eye Hospital, Madurai between July 1993 and February 2001. The ocular and systemic examination details were recorded. RESULTS: Both eyes were affected in 41 (89%) patients. Cataract was present in 81 (93.1%) eyes; most of them were nuclear cataract (79, 97.5%). Other common ocular presentations included microphthalmos in 74 (85.1%) eyes, iris abnormalities in 51 (58.6%) eyes, and pigmentary retinopathy in 33 (37.9%) eyes. Cataract, microphthalmos and iris hypoplasia was a common combination present in 49 (56.3%) eyes. Systemic manifestations included cardiac anomalies in 23 (50%) and neurological anomalies in 16 (34%) children. Multi-system involvement was present in 32 (70%) children. Low birth weight (below 2 kg) was seen in 30% infants. CONCLUSION: CRS may present with a wide spectrum of ocular and systemic findings and requires a high index of suspicion for diagnosis. Any sick infant with unilateral or bilateral congenital cataract should be investigated thoroughly for CRS. PMID- 12532497 TI - Peribulbar anaesthesia for penetrating keratoplasty. A case series. AB - PURPOSE: To prospectively analyse the efficacy and safety of peribulbar anaesthesia for penetrating keratoplasty through a noncomparative, consecutive series. METHODS: One hundred twenty-four (91.1%) of 136 patients undergoing penetrating keratoplasty (PK) from January 1997 to December 2001, were administered peribulbar anaesthesia. The anaesthetic mixture consisted 5 ml of lignocaine, bupivacaine, and hyaluronidase (to avoid evaluation bias) in the peribulbar space. A repeat injection of 3 ml was used if the primary injection was inadequate. Digital ocular compression was done for 10-15 minutes after the first injection. Each patient was analysed for degree of akinesia, subjective patient comfort, analgesia, subjective surgeon comfort, and types of surgical conditions. RESULTS: The age ranged from 19 to 86 years. Forty-nine of 124 patients (39.5%) received PK only and remaining 75 patients (60.5%) received additional procedures. A single injection was sufficient to achieve adequate akinesia (grade II and III) in 114 (92%) patients and 120 (97%) of patients were satisfied (graded pain as < or = grade II). During surgery, 6 (5%) phakic eyes developed episodes of positive intraocular pressure and 5 eyes (4%) developed chemosis. There were no other local or systemic adverse events. The surgeon level comfort was (grade II or more) 98% (122 of 124). CONCLUSION: One-point, low volume, peribulbar anaesthesia for penetrating keratoplasty is safe and efficacions. PMID- 12532498 TI - Frosted branch angiitis associated with rapidly progressive glomerulonephritis. AB - Simultaneous occurrence of frosted branch angiitis and immune-mediated rapidly progressive glomerulonephritis is reported. The two diseases possibly share a common immune mechanism. Patients of frosted branch angiitis should undergo complete systemic evaluation including renal function tests even if the patient is systemically asymptomatic. PMID- 12532499 TI - Detection of cytomegalovirus from vitreous humor in a patient with progressive outer retinal necrosis. AB - A case of progressive outer retinal necrosis (PORN) caused by Cytomegalovirus (CMV), and diagnosed by polymerase chain reaction (PCR) is reported. PMID- 12532500 TI - Congenital hydrocephalus associated with congenital glaucoma and natal teeth. AB - We report the first described association of natal teeth with congenital hydrocephalus and congenital glaucoma, anterior segment dysgenesis with non attachment of the retina. The clinical findings support a diagnosis of Walker Warburg syndrome. The forkhead 7 transcription factor gene is proposed as a candidate gene for this syndrome. PMID- 12532501 TI - Anterior ischaemic optic neuropathy at high altitude. AB - High altitude retinopathy with various manifestations is common after exposure to high altitude. Inadequate autoregulatory response of the retinal vascular system is thought to be responsible for this. We report a case of anterior ischaemic optic neuropathy following exposure to high altitude. PMID- 12532502 TI - Rapid detection of fungal filaments in corneal scrapings by microwave heating assisted Grocott's methenamine silver staining. AB - The Gomoris methanamine silver impregnation technique is a highly reliable and archiveable method of detecting fungal filaments, but the staining procedure is time consuming and laborious. A technique using microwave energy to reduce the duration of Gomori's silver staining is described. PMID- 12532503 TI - Optic nerve entrance coloboma associated with situs inversus. AB - A rare case of optic nerve entrance coloboma associated with situs inversus is reported. PMID- 12532504 TI - Bietti's crystalline dystrophy. AB - This first report from India describes 5 cases of Bietti's crystalline dystrophy without corneal involvement. PMID- 12532505 TI - Use of capsular tension ring in phacoemulsification. Indications and technique. AB - The capsular tension ring (CTR) was originally introduced to reinforce the zonule in eyes with zonular dehiscence and to prevent capsular phimosis in eyes at risk for postoperative capsular shrinkage. Since then, other designs and applications have been developed and described. Modified CTRs with shields can be used in eyes with iris coloboma. A CTR with loops is intended for scleral fixation. Studies are underway to determine if the presence of the CTR helps avoid or limit capsular opacification. A number of different insertion techniques with the aid of manipulators, injectors and traction sutures have been described. We have developed a technique that is particularly effective in cases of zonular dehiscence due to its low solicitation to the damaged bag. PMID- 12532506 TI - Interpretation of computed tomography imaging of the eye and orbit. A systematic approach. AB - Computed tomography (CT) has revolutionised the diagnosis and management of ocular and orbital diseases. The use of thin sections with multiplanar scanning (axial, coronal and sagittal planes) and the possibility of three-dimensional reconstruction permits thorough evaluation. To make the most of this technique, users must familiarize themselves with the pertinent CT principles and terminology. The diagnostic yield is optimal when the ophthalmologist and radiologist collaborate in the radiodiagnostic workup. In this article we describe a systematic approach to the interpretation of ocular and orbital CT scans. PMID- 12532507 TI - Awareness of diabetic retinopathy among diabetics. PMID- 12532508 TI - Ocular involvement in leprosy. PMID- 12532509 TI - Wide-angle vitreous surgery without stereoscopic diagonal inverter. PMID- 12532510 TI - Indocyanine green enhanced maculorhexis in macular-hole surgery. PMID- 12532511 TI - Enhancing eye donation. PMID- 12532512 TI - On how to create motivation in AIDS eye care. PMID- 12532514 TI - [Lymphoma, leukemia, anemia, autoimmune diseases. Healing from the umbilical cord]. PMID- 12532513 TI - [Tachycardic arrhythmias. Emergency--or is the heart only playing tricks?]. PMID- 12532515 TI - [Osteoporosis. How long should it be treated?]. PMID- 12532516 TI - [Contaminated endoscopes. With my own instruments I would allow myself to be examined anytime]. PMID- 12532517 TI - [Time pressure--libido problems--money shortage. Why there is a crisis in many physicians' marriages (interview by Dr. med. Bernhard Maulen)]. PMID- 12532518 TI - [Child is inattentive, cannot sit still, disturbs the classroom. Is it really a hyperkinetic disorder?]. AB - The clarification of attention-deficit/hyperactivity disorder (ADHD) requires an unequivocal categorization of the diagnostic criteria and a differentiation of the condition from normal variations and other disorder patterns. The ICD-10 criteria comprise the main symptoms inattention, hyperactivity and impulsivity. In kindergarten and preschool children, hyperactivity predominates, while in schoolchildren poor attention and cognitive impulsivity tend to be to the fore. In adolescence, aggressiveness and dissocial behavior coupled with drug abuse become central. In adulthood, ADHD is associated in particular with a tendency towards a lower social status and more frequent delinquency. Establishment of the diagnosis of ADHD requires information from the person to whom the individual relates. In this connection, exploration schemas, check lists and questionnaires are often very helpful. With regard to the differential diagnosis, mental disturbances due to physical causes (e.g. hearing disorders, epilepsy, etc.) must be excluded. Consideration must also be given in particular to comorbidities- found in up to two-thirds of cases--in which disordered social behavior, affective disorders and anxiety disturbances dominate. PMID- 12532519 TI - [ADHD as a risk factor for delinquency and personality disorders? Recognizing the patient at risk]. PMID- 12532520 TI - [Multimodal therapy concept in hyperkinetic disorder. Drugs alone are not enough]. AB - For the treatment of attention deficit/hyperactivity disorder, both medical and behavioral therapeutic concepts have been shown to be effective. Somewhat problematical, however, is the fact that a large percentage of these children retain residual symptoms that need treating over a longer period of time. The value of a multimodal therapeutic approach (combination of medication and behavioral treatment including counseling of parents, teachers and the patient) remains controversial. Over the long-term, and account being taken of a number of indicators extending beyond the core symptoms, however, the multimodal treatment concept would appear superior to treatment solely with psychostimulants. For effective treatment accurate titration and the recording of the changes occurring under medication are important. PMID- 12532521 TI - [Erectile dysfunction. An important manifestation of autonomic diabetic neuropathy]. AB - In Germany, some 4-6 million men, including 1.2 million diabetics, suffer from erectile dysfunction (ED). Various other diseases including heart disease, hypertension, arteriosclerosis, hyperlipidemia, endocrine disorders, chronic renal insufficiency, prior radical prostatectomy, neurological diseases, trauma and the abuse of alcohol, tobacco, and side effects of medications, are frequently associated with ED. Medical history, clinical examination, routine blood chemistry and sexual hormone levels may help clarify the etiology of ED. Normally, relaxation of the smooth muscles of the corpus cavernosum--mediated by cGMP and cAMP--together with dilatation of penile arteries and occlusion of venous outflow, results in an erection. The oral type V phosphodiesterase inhibitor, Sildenafil, or prostaglandin E1 injection elevates the cGMP and cAMP levels, respectively. Other therapeutic options include mechanical aids, surgery, hormone replacement or sublingual apomorphine. Since 1998, Sildenafil, an effective, simple and safe oral treatment, has been available. PMID- 12532522 TI - [With the right shut. Well prepared for dialysis]. AB - To accomplish hemodialysis in the patient requiring regular dialysis, a well-made shunt establishing a permanent access to the vascular system is essential. This involves making a connection between a vein and an artery, either directly or via a plastic prosthesis. A classic example is the Cimino shunt, a subcutaneous shunt between the cephalic vein and the radial artery, which today can usually be carried out under plexus anesthesia. It is important to establish the indication for a shunt in good time and to optimally prepare the patient, e.g. by vein training. In cooperation with the nephrologist, the vascular status of the patient, and prior diseases such as peripheral occlusive arterial disease or diabetes mellitus, must be established. A fully functional shunt is essential for the patient with terminal kidney failure to be able to lead a civilized life of the long-term. PMID- 12532523 TI - [Early diagnosis and therapy of Alzheimer dementia. Careful documentation prevents degeneration]. PMID- 12532524 TI - [Diagnostic quiz. Paroxysmal dyspnea. Atrial myxoma]. PMID- 12532525 TI - [Fatal decisions from Medical Congress. Internists to be eliminated]. PMID- 12532526 TI - [Antibiotics in routine practice. With proper selection rapid success]. PMID- 12532527 TI - [Azithromycin for pediatric indications. Brief therapy is ideal for children]. PMID- 12532528 TI - [An occasional patient even forgets to remove the protective cap. "Asthma inhalers must be easy to use"]. PMID- 12532529 TI - [Neuropathic pain. Often mishandled for years]. PMID- 12532530 TI - [Lowering blood pressure with sartans. Are there differences?]. PMID- 12532531 TI - [Leukotriene antagonist. All around talent in asthma and allergies]. PMID- 12532532 TI - [The general practitioner is often the first to notice cognitive deficit. Dementia or not? How to decide]. PMID- 12532533 TI - [First i.m., now oral. COX-2 inhibitor in acute lumbago]. PMID- 12532534 TI - [Description of diabetes by Aretaios von Kappadokien. A miserable, difficult life]. PMID- 12532535 TI - [Probiotics in the era of evidence-based medicine. Humbug or significant therapy?]. PMID- 12532536 TI - [Practical tips for probiotics use by Prof. H. Lochs, Berlin. Deep effects]. PMID- 12532537 TI - [Hyperhomocysteinemia, coagulation disorders, inflammations: new risk factors for stroke]. PMID- 12532538 TI - [Urinary incontinence. When the bladder does not cooperate anymore]. PMID- 12532539 TI - [Treatment of warts. An acne ointment helps, too]. PMID- 12532540 TI - [New diagnostic possibilities and medications. Fit for influenza?]. PMID- 12532541 TI - [Diagnostic procedures in chronic heart failure. What is necessary, significant, contraindicated?]. AB - The most common causes of the heart failure syndrome are coronary heart disease, dilated cardiomyopathy, secondary decompensated hypertension and cardiac defects. Apart from the establishment of the diagnosis, essential diagnostic aims are the clarification of the etiology, the identification of reversible causes, the clarification of specific therapeutic options, assessment of severity and evaluation of the prognosis. A standardized diagnostic work-up based on current consensus recommendations, makes good sense. Symptoms and clinical findings have limited sensitivity and specificity. The leading diagnostic procedure is (Doppler) echocardiography. In the case of ischemic cardiomyopathy, the angiographic coronary status and myocardial vitality evaluation are necessary to clarify the revascularisation option. Further diagnostic examinations (invasive hemodynamics, myocardial biopsy, CT/MRI, ergospirometry, markers of neurohormonal activation) should be carried out as dictated by the individual situation. In patients with an increased risk of developing heart failure (e.g. post-myocardial infarction status, essential hypertension, diabetes mellitus) the aim should be the early detection of an asymptomatic ventricular dysfunction by means of echocardiography. PMID- 12532542 TI - [ACE inhibitors, beta blockers, AT-1 antagonists in heart failure. Right dosage and combination]. AB - Medical treatment of chronic heart failure is applied in accordance with the recommendations of the Task Force Report of the European Society of Cardiology [9] adapted to the respective NYHA stage of the cardiac failure. Currently, it includes the use of ACE inhibitors, beta blockers, AT1 receptor antagonists, diuretics including the aldosterone antagonists, and digitalis. While a positive impact on the prognosis has been confirmed for ACE inhibitors, beta blockers and aldosterone antagonists, this is not the case for diuretics and digitalis. These substance groups are used in the treatment of chronic heart failure because of their morbidity-lowering action. The objective of more recent therapeutic concepts is to block neurohumoral achses or local maladaptation processes (e.g. endothelial antagonists, cytokine inhibition, apoptosis inhibition) activated during the heart failure, or to promote protective mechanisms (e.g. endopeptidase inhibition). Here, however, the results of ongoing or planned randomized studies have to be awaited. PMID- 12532543 TI - [Psychosocial factors in chronic back pain]. AB - The evolution of the industrial era into the information society brought with it a dramatic increase in back pain. Epidemiological data are at odds with the former concept of somatic overload and degeneration as the underlying causes of back trouble. Personality, as well as psychosocial and socio-economic factors all impact on the evolution from acute to chronic back pain. It is suggested that the model of external causes and linear responses to explain back pain will be replaced by the holistic, nondeterministic complexity concept, i.e. multifactorial etiopathogenesis, multidimensional interactions, nonlinear causal pathways. PMID- 12532544 TI - [The weary of life often see their family physician before taking the last step. 16 questions to assess the suicide risk]. PMID- 12532545 TI - [Open trial using ACE inhibitor. Left ventricular hypertrophy diminishes]. PMID- 12532546 TI - [Ezetimib--the first selective cholesterol resorption inhibitors. Strengthening of statins]. PMID- 12532547 TI - [Ezetimib plus statin combination. A strong duo]. PMID- 12532548 TI - [Pegylated interferon alpha-2a. Quality of life improves significantly during successful therapy]. PMID- 12532549 TI - [Chronic hepatitis C from the viewpoint of the patients. There are many causes for the quality of life of patients with chronic hepatitis C to be impaired. But there also exist many possibilities to change it]. PMID- 12532550 TI - [Cholesterol resorption inhibition prevents high statin doses. Lipid lowering in double-pack]. PMID- 12532551 TI - [Fewer fractures after just 6 months. Fast help for tired bones]. PMID- 12532552 TI - [Acid coxibs, NO donors, cytokine inhibitors. New beginnings against rheumatism]. PMID- 12532553 TI - [Osteoporosis: prevention of bone loss and fractures]. PMID- 12532554 TI - [Attachment representation of adolescents in residential care]. AB - In this investigation the attachment representations of adolescents in residential care were examined for the first time. 72 adolescents were interviewed by using the Adult Attachment Interview (AAI). At the same time the degree of adolescent psychopathology was recorded. For this purpose the caregivers completed Achenbach's Child Behavior Checklist (CBCL), the adolescents themselves answered Achenbach's Youth Self Report (YSR). The adolescents in this sample proved to be severely burdened in psychopathological terms. They had access to only an insecure and, in many cases, an extremely insecure attachment representation. For a sub-group of adolescent mothers the early infant-mother attachment was examined using Ainsworth's Strange Situation. The findings show an intergenerational transmission of insecure attachment relationships. The implications of these results for the practice of residential care inspired by attachment theory are discussed. PMID- 12532555 TI - [Preventing violence in schools--approaches and outcomes]. AB - The article gives an overview of the development and evaluation of violence prevention efforts realized in and for schools. The different approaches are divided into structural and organizational measures, the promotion of violence prevention competencies of teachers, violence prevention programs for pupils, and multimodal programs. These approaches are analyzed with respect to contents and methods. Finally research needs for future evaluation studies on violence prevention measures are outlined. PMID- 12532556 TI - [Treatment techniques in systemic family therapy with children]. AB - Play and creativity are essential ingredients of therapeutic work with children. Systemic family therapists have to take the developmental stage of children into account and must adapt their interactional style, the therapeutic setting, their language and their interventions to the needs of children. Since the very beginning of family therapy, a large number of practical recommendations for child participation in family therapy have been compiled. This article presents a number of therapy techniques and case examples illustrating the broad range of interventions that are being applied in systemic family therapy with children. PMID- 12532557 TI - [The placebo: an old problem of methodology revisited]. PMID- 12532558 TI - Drug-induced hepatotoxicity. PMID- 12532559 TI - [Placebo in clinical research--a continual compromise between ethical requirements and scientific rigor]. AB - The use of placebo in clinical trials has been the object of continuous controversy. If effective treatment is not available, then the comparison of a new treatment versus placebo is the only sound and ethical approach. If effective treatment exists and its withdrawal can cause severe or irreversible harm or severe suffering, the use of placebo is forbidden. In all other instances, the scientific community has been divided into two extreme positions: the placebo orthodoxy considers the use of placebo as the only scientifically and hence ethically valid approach; and the active-control orthodoxy denies the use of placebo and recommends the comparison between active treatments. The latter has been supported by the Declaration of Helsinki that, until the Edinburgh 2000 Version, denied the use of placebo whenever effective treatment existed. This position has been the object of recent, strong criticism by ethicists and international Institutions, to the point that in October 2001 the World Medical Association decided to review paragraph 29 of the Declaration. The new version of the Declaration admits the use of placebo, even in the presence of effective treatment, when there are scientifically sound methodological reasons or when the patients receiving placebo are not exposed to any additional risk of serious or irreversible harm. The problem now is that the sudden opening of the World Medical Association may be interpreted as an encouragement to use placebo. Thus, it is necessary to define a new area for a scientifically motivated, but not unscrupulous use of placebo. PMID- 12532560 TI - The Alstrom syndrome: is it a rare or unknown disease? AB - The Alstrom syndrome is a rare, autosomal recessive disorder characterized by retinal degeneration, obesity, progressive hearing impairment, non-insulin dependent diabetes mellitus and kidney and heart failure. Mental retardation is absent and the extremities are normal. The Alstrom syndrome gene located on chromosome 2, has been recently identified. The Alstrom syndrome involves multiple organ systems with a complex interaction between pathways. Phenotypic expression varies considerably, even within sibships. Manifestations observed in some, but not all, Alstrom syndrome patients include acanthosis nigricans, alopecia, short stature, scoliosis, kyphosis, hyperostosis frontalis interna, muscle dystonia, advanced bone age and subcapsular cataract. Other metabolic and endocrinological abnormalities have been described: hypothyroidism, hypogonadism, diabetes insipidus, growth hormone deficiency, hyperuricemia and hyperlipidemia. In the final stages of the disease, affected individuals exhibit progressive chronic nephropathy with eventual kidney failure. The most frequent causes of death include hepatic dysfunction and congestive heart failure secondary to dilated cardiomyopathy. We have summarized our personal clinical data and the information from the scientific literature on the topic in order to provide an up to-date review on the Alstrom syndrome. PMID- 12532561 TI - [Cystic fibrosis in adults]. AB - In the last two decades the life expectancy of patients with cystic fibrosis (CF) has improved enormously: it is now estimated at around 30 years, but some patients reach an older age and one third of all patients with CF currently reach adulthood. Once considered a pediatric disease, CF now is a disease of adults too. As a consequence, internists must be highly knowledgeable about all the clinical patterns of this multifarious disease, must be better qualified to recognize its complications and must be highly trained to treat them. This review is dedicated to internists and general practitioners and not to the specialists in the research and clinical field of CF, until now only pediatricians; it briefly summarizes the medical history of CF and the current knowledge at the molecular, genetic and clinical levels about this disease; it reports up-to-date epidemiological data and it illustrates the clinical patterns which are more frequently encountered in adult patients. The true target of this review is to stimulate a greater interest and participation in this clinical field by adult caregivers, as a development of a medical care system for adults with CF is now crucial. Therefore, a growing number of internists will be increasingly involved in providing care for patients with this disorder. PMID- 12532562 TI - Evaluation of liver fibrosis in chronic hepatitis C with a computer-assisted morphometric method. AB - Objective methods are needed to quantitatively assess the burden of fibrous tissue in liver biopsy specimens and its changes after treatment. The aim of this study was to assess the validity of a computer-assisted morphometric method in the evaluation of liver fibrosis in patients with chronic hepatitis C. Sixty-nine liver biopsy specimens stained with Sirius red were evaluated by two independent observers with a computer-assisted morphometric method to measure the percentage of fibrous tissue in the optic fields examined (fibrosis ratio). Furthermore, 11 pairs of liver biopsy specimens obtained before and after treatment from patients with chronic hepatitis C were evaluated with morphometry by two independent observers in order to assess in which direction fibrosis changed. In the 69 patients, the correlation of the morphometry-measured fibrosis ratio pairs by the two observers was high (r = 0.781). However, the differences between paired values were large, reaching +/- 5% in 95% of instances. The fibrosis ratios observed with morphometry by the two examiners correlated poorly with the Ishak's staging score. The two examiners agreed in 10 out of 11 instances in judging in which direction fibrosis had changed. In conclusion, using our present technique of computer-assisted morphometry, the quantitative assessment of the percentage extension of fibrous tissue was not sufficiently accurate. However, computer assisted morphometry proved to be useful when evaluating the direction of fibrous changes in pairs of liver biopsy specimens from patients with chronic hepatitis C before and after treatment. PMID- 12532563 TI - AMIL survey (anemia in Internal Medicine in Liguria). AB - The aim of the present study was to evaluate the epidemiology of anemias in Internal Medicine Departments. The study involved all patients discharged between May 1 and June 15, 2001 who, at the time of admission, had presented with hemoglobin levels < 12 g%. One thousand and nineteen data collection forms were deemed suitable for analysis and were divided according to the etiology and severity of the anemia. We discovered that simple diagnostic tests (reticulocyte count) were underused, that the majority of anemias were not corrected during hospitalization and that most inpatients of Internal Medicine wards in our Region are elderly (mean age 75 years) and suffer from polypathologies. We believe that further observational studies should be carried out in order to evaluate the progress of patients with anemia and to establish guidelines for the diagnosis and treatment. PMID- 12532564 TI - [Cholestasis from ticlopidine: two clinical cases]. AB - The development of cholestasis during treatment with ticlopidine is an uncommon event in view of the large number of patients to whom this drug is prescribed. We describe the cases of 2 patients who had been treated with ticlopidine for 4 weeks and 2 months respectively. The first patient developed cholestasis with jaundice and pruritus while the second presented with biochemical cholestasis but no jaundice. When treatment with ticlopidine was withdrawn, in the first case pruritus disappeared and jaundice improved. In both cases the liver function tests returned to normal. Both patients were treated with ursodeoxycholic acid which might have an anticholestatic effect in case of hepatotoxicity induced by ticlopidine. PMID- 12532565 TI - [Three cases of bartonellosis (cat scratch disease)]. AB - In the last years the number of Bartonella species has conspicuously increased, with better clinical and diagnostic definitions of the different related illnesses. We report, in the present article, 3 cases of cat-scratch disease, a particular bartonellosis mainly caused by Bartonella henselae which is transmitted by the cat. After an incubation period, cat-scratch disease clinically manifests as a regional lymphoadenitis, with the appearance of a small cutaneous lesion at the site of inoculation of the germ. The pathogenetic agent is susceptible to a number of antibacterial agents, such as penicillin, cephalosporins, aminoglycosides, tetracyclines, macrolides, quinolones, trimethoprim combined with sulphamethoxazole and rifampicin. The prognosis is usually favorable. PMID- 12532566 TI - [Hepatic toxicity of propafenone: a case description]. AB - A case of acute liver injury associated with the use of the antiarrhythmic drug propafenone in a 62-year-old woman undergoing clinical observation for recurrent atrial fibrillation is reported. Propafenone hydrochloride, a class 1C antiarrhythmic drug widely used in the clinical practice for the treatment of supraventricular and ventricular arrhythmias, rarely (0.1-0.2% of incidence) causes liver injury characterized by a rise in hepatic cell enzymes or cholestatic enzymes or both. Within 2 months of the discontinuation of therapy the liver function tests return to normal, therefore there are no known fatalities secondary to propafenone liver injury including fulminant hepatitis and death. The close temporal relationship between the administration of the drug and the acute onset of signs of liver injury, the marked rise in liver function tests following the increase of the drug dosage and their gradual normalization after its withdrawal strongly suggest that propafenone is involved in the pathogenesis of this syndrome. Although rare, hepatotoxicity due to this widely used antiarrhythmic drug should be borne in mind in the differential diagnosis of sudden hepatocellular or cholestatic syndrome of obscure origin. It seems prudent to obtain baseline liver function before starting therapy with propafenone and then follow up laboratory tests some months later at least in patients with known liver disease. PMID- 12532567 TI - [Post-graduate education in internal medicine in the United Kingdom: issues for comparative evaluation]. AB - The structure and the organization of Internal Medicine post-graduate training in the United Kingdom are described. Those aspects that differentiate the British and the Italian post-doctoral training in Internal Medicine are highlighted. Considerations are made about the different nature of an analytic British education as compared to the synthetic approach of the Italian curriculum. The widespread United Kingdom policy of an Internal Medicine approach to clinical training before entering the medical sub-specialty practice is also underscored. PMID- 12532568 TI - [Blastocytosis. An Italian story]. PMID- 12532569 TI - Tobacco control in Pakistan: a challenge for the medical profession. PMID- 12532570 TI - Smoking prevalence and awareness about tobacco related diseases among medical students of Ziauddin Medical University. AB - OBJECTIVE: To determine the prevalence of smoking and awareness of tobacco related diseases among medical students. METHODOLOGY: A survey of all medical students of Ziauddin Medical University in the year 2000. The data was collected by a self-administered questionnaire. RESULTS: A total of 264 (92%) out of 288 students responded to the survey. Smoking was more prevalent among males (26%) as compared to female students (1.7%). The knowledge about tobacco related diseases regarding lung cancer and stroke was significantly higher among females, while knowledge about small for date babies was significantly higher in males. CONCLUSION: Smoking prevalence was higher among male students of Ziauddin Medical University, a trend similar to other medical colleges. Further studies regarding risk factors of smoking will help us to better understand the complex etiology of smoking related disorders. PMID- 12532571 TI - Comparative study--efficacy, safety and compliance of intravenous iron sucrose and intramuscular iron sorbitol in iron deficiency anemia of pregnancy. AB - OBJECTIVE: (1) To determine an alternative iron supplementation with better efficacy, compliance & safety in treatment of iron deficiency anemia during pregnancy, (2) to reduce blood transfusion during pregnancy, labor and puerperium. MATERIAL AND METHOD: A prospective comparative study. A total number of 60 pregnant women with the gestational age of 12-34 weeks were included in the study who were suffering from iron deficiency anemia. They were divided in 3 groups (A, B and C). Group A (n = 15) received intravenous iron sucrose according to recommended dose containing 500 mg of iron sucrose for storage, in group B (n = 20) iron sucrose was administered according to deficit calculated as per formula but 200 mg of iron was given for storage instead of 500 mg, to reduce cost. While group C received intra muscular iron Sorbitol in the dose used as practice. RESULTS: Mean hemoglobin in group A and B was 8.0 +/- 1.1 g/dl and 8.9 +/- 0.7 respectively, in group C, it was 8.8 +/- 0.9 g/dl. In group A & B initial hemoglobin was assessed 3 weeks post therapy which showed an average rise of 2.8 g/dl (group A) and 1.9 g/dl (group B) and second assessment of Hemoglobin was done prior to delivery (ave: 6.6 weeks) showed a total rise of 3.8 g/dl (group A) and 2.4 g/dl (group B). Pre delivery mean Hemoglobin in group A and B was 11.8 g/dl and 11.3 g/dl respectively. In group C, the Hemoglobin was assessed only prior to delivery (average: 8.4 weeks from the start of therapy), and a rise of 1.4 g/dl was observed with pre delivery mean Hemoglobin of 10.2 g/dl. Target hemoglobin levels i.e. 11 g/dl was achieved by 80% in Group A, 70% in Group B and 28% in Group C by the time of delivery. Blood transfusion was not required in any group. In group A and B one patient had moderate abdominal pain, 2 had weakness and shivering and 3 had phlebitis at the site where intravenous canula was retained. None of patient discontinued the therapy due to any adverse effect. In group C majority complained of pain at injection site while 5 patients dropped out from the study due to intolerance. CONCLUSION: Intravenous iron therapy is safe, convenient and more effective then intramuscular iron therapy in treatment of iron deficiency anemia during pregnancy. The intravenous iron therapy can replace blood transfusion in antenatal period. PMID- 12532572 TI - Transitional cell carcinomas of the urinary bladder. A histopathological study. AB - OBJECTIVE: To determine the histological grading and muscle invasion in Transitional Cell Carcinomas of the Urinary Bladder; and to evaluate whether any correlation exists between tumour grade and muscle invasion. METHOD: A Retrospective study of all consecutive cases of Transitional Cell Carcinomas of the Urinary Bladder diagnosed at Aga Khan University Hospital between 1st Jan 1997 and 31st Dec 2000. RESULTS: This study included 495 cases of Transitional Cell Carcinoma of the Urinary Bladder. M: F ratio was 4:1. Ages of patients ranged from 30 years to 87 years. Mean age in males was 59.1 years and in females 58.8 years. Forty four percent cases were Grade II, and 29.5% Grade III. None of the Grade I lesions were invasive, 10% Grade II tumours, 61.41% of Grade III and all Grade IV tumours were invasive. CONCLUSION: There is a definite correlation between advancing tumour grade and muscle invasion. PMID- 12532573 TI - Seroprevalence of hepatitis B, C and HIV in blood donors in northern Pakistan. AB - OBJECTIVE: To study the seroprevalence and trends of Hepatitis B, C and HIV sero markers in healthy blood donors of Northern Pakistan. SUBJECTS AND METHODS: Blood donated by healthy donors from both Armed Forces and civilian population, collected from Jan 1996 to Dec 2000 were tested by Enzyme Linked Immunoassay at Armed Forces Institute of Transfusion Rawalpindi, Pakistan. Demographic data of these donors was also studied. RESULTS: Of 103858 blood donors, 3.3% (95% CI 3.20%-3.41%) were HBsAg, 4.0% (95% CI 3.91%-4.11%) were anti HCV and 0.007% anti HIV positive. Their average was 28 years. HBsAg positive donors were a decade younger than anti HCV positive donors. Pattern in Armed Forces and civilians donors was similar but there was significant reduction in the prevalence of HBsAg carriage in all blood donors. CONCLUSION: This study supports the changing trends in HBV/HCV seroprevalence in blood donors and a low prevalence of HIV in Pakistani population. PMID- 12532574 TI - Urbanisation and activity pattern of south Asian children. AB - OBJECTIVE: To compare the physical activity level and total energy expenditure of 10-12 year old school children living at different levels of urbanization. METHOD: All the participating children kept a three-day record of their activities, for every fifteen-minutes, in specially designed diaries. Activities were grouped according to intensity that was determined on the basis of Physical Activity Ratio (PAR). After calculating the average time spent in a day, in activities of varying intensity, overall Physical Activity Level (PAL) of each subject was calculated by factorization method. After measuring bodyweight using a standard equation Basal Metabolic Rate (BMR) of the subjects was calculated. Total Energy Expenditure (TEE) was assessed on the basis of PAL and BMR of each child. SUBJECTS: Physical activity level of six groups of 10-12 year old children, representing various urbanization categories, was studied. Three groups of children were recruited from Punjab, Pakistan: rural, middle income urban and high income urban, and they were assigned urbanization rank (UR) 1, 2 and 3. Another three groups of children were recruited from Slough, UK: British Pakistani, British Indian, and British Caucasian and they were assigned urbanization rank 4, 5 and 6 respectively. RESULTS: Physical activity level decreased significantly with the urbanization rank only among girls of Pakistani origin (UR 1-4). Pattern of gender differences in activity level was different in rural and urban children. Rural girls were slightly more active than rural boys, whereas in urban areas boys were significantly more active than girls. Because of lower bodyweight the less urbanized children in spite of having higher PAL had a lower mean TEE as compared to the more urbanized groups. CONCLUSION: The activity level of rural children having access to formal education and television could not be expected to be very different from their urban counterparts. Inactivity of urban girls needs particular attention. Because of lower caloric requirements, on similar diets, less urbanized groups may succumb to overweight more easily than the urbanized groups. Participation in active games may present a substitute to decreased involvement in moderately active work and play activities. PMID- 12532575 TI - Susceptibility patterns of Escherichia coli: prevalence of multidrug-resistant isolates and extended spectrum beta-lactamase phenotype. AB - OBJECTIVE: To study Escherichia coli (E. coli) susceptibility, prevalence of multidrug-resistant isolates and possible role of extended spectrum beta lactamases (ESBL) in E. coli resistance. SETTING: Department of Medicine, Shifa College of Medicine, Islamabad. METHODS: Three hundred and seventy eight E. coli isolates from different sources were identified during six-month period. Susceptibility to various antibiotics was checked using standard methods. Multidrug-resistant isolates were separated. Isolates resistant to cefpodoxime and aztreonam were considered as ESBL phenotype, indicating ESBL production. RESULTS: Most of isolates were resistant to ampicillin and trimethoprim/sulfamethoxazole. Overall resistance to ciprofloxacin, ceftriaxone, cefotaxime and cefpodoxime was 49%, 34%, 36% and 38% respectively. Resistance to fourth generation cephalosporins was lower. Almost all Isolates were sensitive to amikacin, imipenem and tazobactam. Aztreonam resistance was found in 25% isolates. Similar pattern was observed for urinary E. coli isolates; 45% E. coli isolates were found to be multidrug-resistant. Nitrofurantoin showed low-level resistance both to multidrug-resistant as well as urinary isolates. Resistance to both cefpodoxime and aztreonam was found in 25% isolates suggesting ESBL production. CONCLUSION: E. coli resistance in Pakistan is much higher than reported from western literature. Multidrug-resistant isolates, including third generation cephalosporins and quinolones, are very common. ESBL production may contribute to this high level resistance against beta-lactams. PMID- 12532576 TI - Leiomyosarcomas: analysis of clinical presentations in 6 patients. AB - OBJECTIVE: To identify characteristic features of leiomyosarcomas in an attempt to diagnose them preoperatively. STUDY DESIGN: The 3 years experience with 6 patients with uterine leiomyosarcomas at Hayatabad Medical Complex, Peshawar was retrospectively reviewed. RESULTS: The mean age of occurrence was 50 years with abnormal uterine bleeding the commonest symptom. Leiomyosarcomas had no preferential uterine location and were of variable sizes. Only 1 case was diagnosed preoperatively. CONCLUSION: We have observed that physical examination and imaging techniques are rarely helpful in establishing the diagnosis preoperatively. We recommend larger prospective studies to evaluate the efficacy of preoperative monitoring of enlarged uteri. PMID- 12532577 TI - Diagnosis of typhoid fever by Dot enzyme immunoassay in an endemic region. AB - OBJECTIVE: To compare the results of Dot Enzyme Immunoassay (EIA) and concomitantly performed blood culture for the diagnosis of typhoid fever. METHODS: The results of Dot Enzyme Immunoassay (EIA) of patients who also had blood cultures/bone marrow culture performed at the same time were analyzed. The period of study spans between April 1997-September 1997. RESULTS: A total of 1760 Dot Enzyme Immunoassays were performed from April 1997 to September 1997. Of these, only 128 (7.2%) cases had concomitantly performed blood/bone marrow cultures. Therefore, the study was narrowed down to these 128 cases. Sixty-nine cases were culture positive for S. typhi. Of these culture proven cases, IgM antibodies by Dot EIA were positive only in 49 (71%) cases while 20 (28.9%) of culture proven cases remained negative for IgM antibodies. CONCLUSION: Taking blood/bone marrow culture as gold standard, the sensitivity of Dot EIA was found to be 71% while specificity was found to be 43%. The predictive value of the positive test was found to be 59.7% and that of a negative test was found to be 56.5%. PMID- 12532578 TI - Ischemic stroke in young adults of South Asia. AB - OBJECTIVE: To study the risk factors, etiology and management of ischemic stroke in young adults in a South Asian population. METHODS: Retrospective study conducted at a large tertiary hospital in Karachi. One hundred and eighteen patients between fifteen and forty-five years admitted over a five year period with a diagnosis of ischemic infarct constituted the study population. The study variables included the full clinical spectrum, spanning historical, laboratory, radiological and outcome parameters. RESULTS: Forty-three percent of patients were hypertensive and 30% were diabetic. The combination of diabetes and hypertension was found in 19.5% of patients, intra-cranial and carotid artherosclerosis in 22% and embolism in 11% of the cases. The in-hospital mortality was 11%. The outcome was excellent in 27%, good in 50% and poor in 23% of patients. CONCLUSION: The risk factors for artherosclerosis and the contribution of intra and extra-cranial artherosclerosis were found to be much higher than those from the Western Hemisphere. PMID- 12532579 TI - Determination of cerebrospinal fluid sugar by glucometer. PMID- 12532580 TI - Role of imaging in the evaluation of renal trauma. PMID- 12532581 TI - Ectopic pregnancy after tubal sterilization. PMID- 12532582 TI - Histiocytosis X of the vulva. PMID- 12532583 TI - Congenital absence of IVC with azygous continuation. PMID- 12532584 TI - View box case--9. Bronchiectasis. PMID- 12532586 TI - Deep vein thrombosis at high altitude. PMID- 12532585 TI - Prevalence of depression in patients with coronary artery disease in a tertiary care hospital in Pakistan. AB - OBJECTIVE: To determine the prevalence of depression in patients with coronary artery disease (CAD) in a tertiary care hospital setting in Pakistan. METHODS: One hundred and fifty four patients of CAD (115 males and 39 females) were randomly selected from the outpatient department and wards of the National Institute of Cardiovascular Diseases, Karachi and were scored for depression via the Hospital Anxiety and Depression Scale. Basic demographic data and disease variables were also collected. RESULTS: The point prevalence of depression in the sample was 37% (31.3% males and 53.8% females). Female sex, income level below Rs. 5000 per month, low education level, outpatient, single earning family member and hypertension were few variables associated positively with depression (p < 0.05). Only one patient was receiving treatment for depression by his cardiologist. CONCLUSION: Depression is prevalent in CAD patients in Pakistan. Economic conditions may pose an additional threat on these patients. Treating physicians (especially cardiologists) need to be aware of this co-morbidity so as to be able to diagnose and adequately manage such patients. PMID- 12532587 TI - [Fundamental and applied aspects of rehabilitation medicine in neurology]. PMID- 12532588 TI - [Drug electrophoresis: history, achievements and perspectives of development]. PMID- 12532589 TI - [The role of control over microcirculation in millimetric wave therapy of acute destructive pancreatitis]. AB - Microcirculatory reaction to millimetric waves (5.6, 7.1 and 4.9 mm) was studied in 78 patients with acute destructive pancreatitis using laser doppler flowmetry. The 4.9 mm waves proved most effective. As such waves have a normalizing effect on capillary blood flow, they can be used at early stages of the disease. The method allows control of individual reaction of the patient to millimetric waves and, therefore, facilitates the decision on its application. PMID- 12532590 TI - [Efficacy of different methods of ozone therapy in vascular complications of diabetes mellitus]. AB - 21 patients with diabetes mellitus (DM) type 1 and 97 patients with DM type 2 (a total of 118 diabetics) complicated by angiopathy of the lower limbs and diabetic retinopathy (n = 96) received sugar reducing drugs and ozone-oxygen therapy (OOT). Three techniques of OOT were used: external ("ozone boot" method), systemic (intravenous introduction of ozone-inriched physiological solution or rectal insufflations of ozone-oxygen mixture), combined (external + systemic). The external and combined methods proved superior in trophic disorders in the region of distal parts of the lower limbs while the systemic and combined methods produced better effect on the other signs of diabetic angiopathy and retinopathy, functional and biochemical parameters. PMID- 12532591 TI - [Psychological predictors of rehabilitation efficiency of patients with radiculopathy after neurosurgical correction of compression diskogenic syndrome]. AB - Sixty four osteochondrosis patients after neurosurgical correction of the compression syndrome were examined psychologically before and after the course of rehabilitation. It was established that considerable relief of pain in the course of physiotherapy was seen in patients who, before the treatment, had less severe impairment of the psychological status, no significant symptoms of hypochondria, anxiety, depression. Psychologically unfavourable predictors of low efficiency of physiotherapy were psychic tension, excitability, low stress resistance, pessimism, high anxiety, intrapersonality conflicts. PMID- 12532592 TI - [Epidural long-term local pharmacotherapy in patients with degenerative dystrophic spinal diseases during combined rehabilitation therapy]. AB - Epidural long-term local pharmacotherapy in degenerative-dystrophic diseases of the lumbar spine proved most effective in combination with rehabilitation procedures. Such combined treatment can be used in cases with long-standing disease resistant to conservative treatment. PMID- 12532593 TI - [Combined oxygenobaro- and SWF-therapy of patients with traumas and sequelae of elbow joint injury]. PMID- 12532594 TI - [Sanatorium stage of rehabilitation of patients operated for infantile cerebral palsy]. PMID- 12532595 TI - [Effect of low intensity infrared laser radiation on cell membrane lipids in experiment]. PMID- 12532596 TI - [Comparative efficacy of various massage techniques in the rehabilitation treatment of patients with inhalation trauma early after admission to the hospital]. AB - 67 patients with inhalation trauma of 90 patients with acute thermal trauma and 23 healthy controls participated in the trial. External respiration function was assessed on the spirograph SPIROSIFT-3000 (Fukuda Densy, Japan). The injured patients have undergone chest vibromassage (n = 23), vibromassage of Zakhariin Head zones (trachea-bronchi-lungs) (n = 22), manual chest massage (n = 22). Improvement of clinical and external respiration parameters was more significant in patients exposed to vibromassage of the trachea-bronchi-lungs zones. PMID- 12532597 TI - [Ximedon-electrophoresis in rehabilitation of patients with systemic scleroderma]. AB - A double blind placebo-controlled trial performed in 56 patients with scleroderma systematica showed effectiveness of a domestic drug ximedon--a pirimidine compound--applied during electrophoresis on the affected skin, limbs. The addition of ximedon-electrophoresis to rehabilitation program for scleroderma systematica improved the condition in 77.8%, microhemocirculation in 72.2%, reduced the area of the affected skin by 9.8% (p < 0.05), skin induration in 55.6% of patients. PMID- 12532599 TI - [One more comment on syndrome-pathogenetic classification of physiotherapy]. PMID- 12532598 TI - [Utilization of ultrahigh frequency low-intensity electromagnetic radiation in the treatment of cancer patients]. PMID- 12532601 TI - [Application of plasmapheresis in rehabilitation of patients after coronary bypass grafting in low mountain climate]. PMID- 12532602 TI - [Effect of communication with dolphins on children suffering from functional stuttering treated in sanatorium]. PMID- 12532603 TI - [Exercise therapy in the treatment of children with atopic dermatitis]. PMID- 12532604 TI - [Radon capsule for therapeutic application]. PMID- 12532605 TI - [Cryomassage in physical conditioning of children]. PMID- 12532606 TI - [Medical and economic significance of prophylaxis of unfavorable reactions to weather]. PMID- 12532607 TI - [Physiological preconditions for the use of hypoxia in the treatment of gastrointestinal diseases]. PMID- 12532608 TI - Occupational disease in Connecticut, 2000. AB - BACKGROUND: Occupational illnesses in Connecticut are tracked to better understand patterns and trends by industry, cause, and time, which allows for interventions such as cluster investigations and education. METHODS: Data were collected for occupational illnesses from three data sources: the Connecticut Bureau of Labor Statistics (BLS) annual survey, workers' compensation reports, and physician reports. FINDINGS: There were 6,396 reports of occupational illness in Connecticut in 2000. Reports increased sharply based on two of the three databases. Reports were dominated by musculo-skeletal disorders (MSD), followed by lung conditions. Manufacturing, the State of Connecticut, and municipalities tended to have the highest number of cases and rates. Acute respiratory conditions were the most common lung condition, followed by occupational asthma and asbestos-related conditions. CONCLUSION: Occupational illnesses are very prevalent in Connecticut and appear to be increasing. Improved diagnosis and reporting are needed to improve patient care and to direct preventive efforts. PMID- 12532609 TI - Neonatologists' opinions regarding resuscitation of extremely premature infants in Connecticut and Rhode Island. AB - OBJECTIVE: To characterize the range of opinions among neonatologists in Connecticut and Rhode Island regarding the gestational age at which they would support parental resuscitation or nonresuscitation requests for infants born at 21-27 weeks. METHODS: Anonymous questionnaire mailed to 70 Connecticut and Rhode Island neonatologists detailing specific resuscitation scenarios and hypothetical guidelines regarding management of premature infants. RESULTS: Response rate was 69%. Nine percent of neonatologists would never reject a parental resuscitation request. Ninety-one percent would reject a resuscitation request at 21 weeks, 67% at 22 weeks, and 11% at 23 weeks. Fifty-three percent would refuse a nonresuscitation request at 24 weeks, 96% at 25 weeks, and 100% at 26 weeks. CONCLUSIONS: The majority of neonatologists would not resuscitate infants born at < or = 22 weeks and would resuscitate infants born at > or = 25 weeks gestational age despite parental wishes to the contrary. PMID- 12532610 TI - Monitoring antifactor Xa activity of the low-molecular-weight heparins in the clinical setting: who, why, how, and when? PMID- 12532611 TI - Inspection, palpation, percussion, and auscultation = connection. PMID- 12532612 TI - Common sense vs fail safe. PMID- 12532613 TI - Sustainable progress: is it possible? PMID- 12532614 TI - [Cyanoacrylate compared with traditional surgery in tension-free incision closure]. AB - OBJECTIVE: To investigate the efficiency of cyanoacrylate in closing tension-free incisions in out-patient surgery compared with traditional suturing methods. PATIENT AND METHODS: A single, blind, clinical study was carried out on 60 patients divided into two groups. Group 1 (n = 30) received one or two drops of cyanoacrylate directly on the incision. Control (Group 2) (n = 32) received traditional nylon suturing. Both groups were evaluated after 21 days of incision closing. Statistical differences in baseline characteristics between study treatment groups were assessed with chi 2 and t tests. All p values are two sided. RESULTS: There was no significant difference between the two groups in age, gender, or incision length. Length of time for surgical incision to close was less in Group I, significant difference p < 0.0001. There was also a significant difference in aesthetics in Group I. There was no significant difference in any remaining variable, such as suture dehiscence and adequate border coaption. CONCLUSION: Our study showed cyanoacrylate to be more efficient and advantageous in closing of incisions in out-patient surgery than traditional suturing methods. Although we do not pretend to substitute these methods, we recommend the use of cyanoacrylate in closing tension-free incisions in certain patients following rules of usage and practiced by experts. PMID- 12532616 TI - [Morbidity and mortality in newborns with omphalocele and gastroschisis anterior abdominal wall defects]. AB - OBJECTIVE: To identify morbidity and mortality in newborns with congenital defects of the anterior abdominal wall. DESIGN: Descriptive, comparative, and retrospective study. PATIENTS: Thirty nine patients with gastroschisis and 26 patients with omphalocele. RESULTS: Median size of the defect in the gastroschisis group was 4 cm. Infants underwent primary closure en 41% of cases. Post-surgical morbidity occurred in 74% of patients with sepsis the main complication in 61.5%. A total of 16.2% died mainly due to acute renal failure and sepsis. In the omphalocele group, median size of defect was 5.5 cm. Primary closure was done in 65% of patients. Complications occurred in 65% of newborns, sepsis was the most frequent complication (46%). Mortality rate was 16.6% related to acute renal failure and cardiogenic shock. CONCLUSIONS: The main causes of morbidity in the two groups were infections and acute renal failure. Mortality rate was similar to that reported in the world literature for gastroschisis and slightly lower for omphalocele. PMID- 12532615 TI - [Immediate neonatal surgery: initial experience in gastroschisis and omphalocele treatment in Northwestern Mexico]. AB - Immediate neonatal surgery (INS) has the purpose of achieving surgical treatment of a congenital malformation in a newborn within the first few minutes after delivery. Since 1993, we have invited 15 pregnant women with sonographic diagnosis of fetal abdominal wall defect to participate in INS. All were treated by means of elective (eight cases) or emergent (seven cases) cesarean section between 33 and 38 gestational weeks. The neonates were immediately operated on the procedure beginning between 17 and 35 min after delivery. Eleven neonates had gastroschisis and primary closure was accomplished in five cases with application of auxiliary plastic patch (45.5%) and in four cases with use of native tissues only (36.4%); nine neonates survived (81.8%), and were able to be fed by mouth at 14 +/- 4.5 days. Four had omphalocele, and in three primary closure and feeding by mouth before 1 week was attained; the fourth neonate died because of coexisting malformations. The present series, even if small, is useful to affirm the safety of a procedure such as INS for both mother and child; it may contribute to improve the results of treatment of fetuses with abdominal wall defects. PMID- 12532617 TI - [Plasmapheresis in Guillain-Barre syndrome]. AB - OBJECTIVE: To evaluate the efficacy of early therapeutic plasmapheresis in severely affected patients with Guillain-Barre syndrome (GBS). PATIENTS AND METHOD: Patients between 16 to 70 years of age, with GBS according to NINCDS criteria, in stage > or = 3 of Huges severity score and with less than 7 days of onset of symptoms were admitted for therapeutic plasmapheresis. All patients completed a total of five sessions of plasma exchange every other day using 25% albumin and saline in a 1:1 proportion. Follow-up was performed for 6 months. Friedman and Wilcoxon tests were done to establish differences within groups. RESULTS: A total of 34 patients were admitted to the study. The time span between onset of symptoms and admission to the study was 4 days (median). Fourteen (41%) required mechanical ventilation. Clinical progression of the syndrome was observed during the first two sessions of plasma-pheresis. Improvement in motor function by Huges score was observed only after fourth session of plasma exchange (p < 0.05). Two patients did not improve, and one died. CONCLUSIONS: Clinical improvement in GBS during early therapeutic plasmapheresis occurs only after the fourth session of plasma exchange. PMID- 12532618 TI - [The neurologic diseases]. AB - Neurologic diseases have important social and economic repercussions. The economic cost related to medications, caretakers, and therapists is evident, as well as the impact the disease may cause on the family. Epilepsy is the most frequent cause of medical care at the out-patient service of the Manuel Velasco Suarez National Institute of Neurology and Neurosurgery (NINN), in Mexico City and the estimated annual prevalence of this disease is 5.9 per 1,000 inhabitants. Parkinson's disease, the most representative of movement disorders, affects 20% of the populating aged > 65 years. Depression and dementia are complex and heterogeneous neuropsychiatry disorders, representing the fifth cause of morbility after accidents, cardiovascular disease, liver cirrhosis and diabetes mellitus. During the last decades, encephalic trauma has been the first cause of mortality: among brain tumors, glioblastomas have the greatest impact. Among infectious disease of the central nervous system (CNS), which are relatively infrequent as compared to gastrointestinal or respiratory infections; viral encephalitis, AIDS-associated toxoplasmosis, and tuberculous meningitis are the most frequent discharge diagnoses at the NINN. The need for a national registration system of neurologic diseases is unquestionable. Such a system would provide accurate data regarding the magnitude of problems related with such disease and help design adequate care and prevention programs. PMID- 12532619 TI - [Chromatin epigenetic modifications in cancer generation]. AB - Chromatin is more than a simple staining structure inside the nucleus. It represents a level of organization that regulates transcriptional activation in eukaryotic genes; this may be through epigenetic modifications such as DNA methylation, histone acetylation, deacetylation and methylation, and chromatin remodeling complexes. Chromatin can have a direct impact on regulating gene expression. Several human diseases arise from alterations of epigenetic regulation, causing change in chromatin structure: cancer is not the exception. The expression of tumor suppressor genes can be epigenetically silenced by DNA methylation of their promotor regions. In other cases such as leukemias, imbalance of histone acetylation and deacetylation can be a determinant event to induce a leukemic phenotype. Finally, improper imprinting can also be associated with neoplastic transformation as in Wilms tumors and sporadic colon cancer. All this evidence supports our particular attention to the study of chromatin structure in cancer. The use of compounds that modify chromatin to improve effects of radio- and chemotherapy will open new horizons in what we call Chromatin Therapy in cancer. PMID- 12532620 TI - [Pancreatic beta-cell neogenesis and regeneration]. AB - OBJECTIVE: To review the literature on beta-cell neogenesis and regeneration, with special interest in substances that regulate such processes. DESIGN: Representative papers were selected through a computer MEDLINE search from 1990 to 2000. RESULTS: Several studies showed that once islets of Langerhans developed from small pancreatic ducts, this process did not continue in normal adult individuals. However, it has been published that new beta-cell formation can occur in vivo in certain experimental models through neogenesis, in which gene pax 4 is extremely important. On the other hand, substances that stimulate the regenerative process of beta-cells include glucose, several hormones and certain growth factors. CONCLUSIONS: Substances that stimulate the regenerative process commonly express this effect in elevated, non-physiologic concentrations; thus, their possible therapeutic importance is not yet clear. Nevertheless, the Reg protein, present in regenerative pancreas and implied in neogenesis process, has actual possibilities of becoming therapeutically important. PMID- 12532621 TI - [Sixty-year-old male with community-acquired pneumonia and respiratory insufficiency]. PMID- 12532622 TI - [Mixed germinal cell tumor in neck associated with additional congenital malformations in a newborn]. AB - The case of a female premature newborn is presented with a tumor in neck and severe perinatal asphyxia that required mechanical ventilation from birth; she died at 28 days dew to heart and renal failure. Surgical resection of cervical tumor was not possible due mechanical ventilation and persistent hemodynamic alteration. Autopsy revealed a mixed tumor of germinal cells, persistence of ductus arteriosus, permeable foramen ovale, congenital, hydrocephalus, dysgenetic corpus collosium, and four lobes in right lung. PMID- 12532623 TI - [Primary infratentorial lymphoma of the central nervous system]. AB - An 39 year-old woman presented with neurotic and neurologic symptoms is described. Computerized tomography revealed contrast uptake in nodular form of oval centers and perilesional edema. Clinic characteristics and possible etiopathogeny are discussed. Post-mortem examination disclosed presence of a primary cerebral lymphoma type B, not involving other organs. PMID- 12532624 TI - [Cremation. A public health chapter in Mexico]. AB - Cremation was a part of the funeral ceremony in prehispanic Mexico. When the Spanish conquerors came, this practice was prohibited. In 1877, the Uppu Health Council authorized animal incineration to avoid consumption by indigents or transformation in putrid emanation. Creamation was welcome in Mexico because of the knowledge of hygienic at the time, whose adepts had evaluated toxic exhalations of cadavers and the cemeteries of Mexico City, with incomplete destruction of the cadaver and filtration of contaminants into the subsoil. Three were the reasons against cremation; religious beliefs, lawmaker medical preoccupation with the disappearance of legal evidence, and the newly born science of anthropology, with loss of material for laboratory. The first crematorium was inaugurated by Dr. Eduardo Liceaga in February 1909 in the Dolores Cemetery. PMID- 12532625 TI - [A cause of costal rib notch (Roesler's sign)]. PMID- 12532626 TI - [New genes involved in the appearance of cancer]. PMID- 12532627 TI - [The combination on conjugated estrogens and medroxyprogesterone in the post menopause patient and risk of breast cancer: analysis of an uncertain statistic]. PMID- 12532629 TI - [A United States National Institutes of Health investigation suspended due to risk of breast cancer with use of hormone replacement in menopause]. PMID- 12532630 TI - [Quantification experience in climacteric and menopause in IMSS physicians]. PMID- 12532631 TI - [Energy expenditure and metabolic state in patients with Guillain-Barre syndrome]. PMID- 12532632 TI - [The treatment of Pseudomonas aeruginosa infections; current status and future strategies]. PMID- 12532633 TI - [Overview of lipophilic yeast Malassezia: the current status of the molecular diagnosis]. AB - The lipophilic yeast, genus Malassezia is a part of the normal cutaneous microflora of human and warm-blooded vertebrates. Species of the genera were re classified to seven species; M. pachydermatis, M. globosa, M. furfur, M. obtusa, M. restricta, M. slooffiae and M. sympodialis. However, the means of species identification in conventional clinical laboratories have not been reported and the clinical significance of each species is not clearly understood. Species identifications of genus Malassezia which depend on the morphological, physiological characters are difficult and time-consuming. Recently, many molecular techniques have been developed for identification or typing of Malassezia. PCR-mediated methods, PCR-direct sequencing and nested-PCR using specific primers, are useful to identify the spices. The basic information obtained from these approaches have been contributing to the understanding of these pathogenic yeasts and related diseases. PMID- 12532634 TI - [A questionnaire survey on the theory of postoperative infection prophylaxis in urology]. AB - A questionnaire survey on the theory of postoperative infection prophylaxis was conducted to obtain the consensus on perioperative antimicrobial use among urologists in Japan in the period from April to July 2000. Sixty-three of the 87 urologists replied, and the following consensus was obtained. An antimicrobial prophylaxis (AMP) agent should be chosen based on their efficacy against the pathogens expected to be contaminants, such as Escherichia coli, Staphylococcus spp., Klebsiella pneumoniae and Bacteroides fragilis group. Use an AMP agent that achieves a bactericidal concentrations in both the serum and operating site. Use an AMP agent that has little unfavourable side effects. The newer agents should be considered as a therapeutics for postoperative infections. The therapeutic antimicrobial agents having no cross-resistance to the AMP agents should be used, if postoperative infection is suspected or developed. The most commonly used agent for clean operations are cefazolin (CEZ), followed by cefotiam (CTM) and piperacillin (PIPC), in this order. For clean-contaminated operations, the most commonly used agent is CTM, followed by PIPC and CEZ. PMID- 12532635 TI - [Antibacterial activities and PK/PD parameters of aminoglycosides against recent clinical isolates of gram-negative rods]. AB - We examined antibacterial activities and PK/PD parameters of six kinds of aminoglycosides against seven bacterial species of clinical isolates in 2001. Aminoglycoseides examined were gentamicin (GM), dibekacin (DKB), tobramycin (TOB), amikacin (AMK), netilmicin (NTL), and isepamicin (ISP), and bacterial isolates used were each 50 strains of Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Citrobacter freundii, Proteus spp., Serratia marcescens and Pseudomonas aeruginosa. All aminoglycosides showed good activities with low MICs against 6 species of Enterobacteriacea except S. marcescens. Eight strains (3.2%) among them were resistant to one or more aminoglycosides. Resistance to multiple aminoglycosides were detected in 16 strains (32%) of S. marcescens, among which 13 strains were resistant to AMK but susceptible to ISP. Three (6%) strains of P. aeruginosa were resistant to multiple drugs, one of which was resistant to all six aminoglycosides, and others were moderately susceptible to AMK and ISP, and susceptible to GM, AMK and ISP. Using a ratio of peak serum concentration to MIC90 (Cmax/MIC90) or a ratio of area under the curve to MIC90 (AUC/MIC90) as a pharmacokinetic and pharmacodynamic (PK/PD) parameter, we estimated the efficacy of the drug. An excellent effect of ISP, which was injected intramuscularly or intravenously at a dose of 400 mg, was expected for strains of Enterobacteriacea except S. marcescens. The Cmax/MIC90 ratios for S. marcescens were comparably higher in GM and ISP and that for P. aeruginosa were rather high in TOB when compared to other aminoglycosides. Another PK/PD parameter, AUC/MIC90 ratio, was high enough in NTL and ISP for Enterobacteriacea, suggesting good efficacy of these drugs. The (AUC/MIC90) ratios for S. marcescens were comparably high in GM and ISP, and that for P. aeruginosa were high in TOB, DKB, and ISP. PMID- 12532636 TI - [Antimicrobial susceptibility of Streptococcus pneumoniae and Haemophilus influenzae isolated in major hospitals in Nagano Prefecture]. AB - We determined the minimum inhibitory concentration (MIC) of various antimicrobial agents against 108 strains of Streptococcus pneumoniae and 144 strains of Haemophilus influenzae isolated from respiratory organs in the First Department of Internal Medicine, Shinshu University, and affiliated hospitals between January 2000 and February 2001. The following results were obtained. 1. Fifty-one (47.2%), 56 (51.9%), and 1 (0.9%) of 108 strains of S. pneumoniae were classified as penicillin-susceptible S. pneumoniae (PSSP), penicillin-intermediate S. pneumoniae (PISP), and penicillin-resistant S. pneumoniae (PRSP), respectively. 2. Three carbapenems had potent antimicrobial activity against PISP and PRSP. Furthermore, none of the strains were highly resistant (MIC > 2 micrograms/ml) to benzylpenicillin, ampicillin (ABPC), sulbactam/ampicillin (SBT/ABPC), cefotaxime (CTX), or cefepime (CFPM). 3. Eleven (7.6%) and 6 (4.2%) of 144 strains of H. influenzae were classified as beta-lactamase-producing ABPC-resistant strains and beta-lactamase negative ABPC-resistant H. influenzae (BLNAR), respectively. Levofloxacin, sulfamethoxazole/trimethoprim, and meropenem had potent antimicrobial activity against these resistant strains. 4. BLNAR strains were more highly resistant to CTX, CFPM, SBT/ABPC, and cefaclor than beta lactamaseproducing strains. 5. In our surveillance study regarding clinical isolates of S. pneumoniae and H. influenzae from respiratory organs in Nagano prefecture, there were regional differences in the isolation rate and antimicrobial susceptibility. The isolation rates of resistant strains were lower than those reported in a nationwide survey. PMID- 12532637 TI - [Susceptibilities of bacteria isolated from patients with lower respiratory infectious diseases to antibiotics (2000)]. AB - From October 2000 to September 2001, we collected the specimen from 410 patients with lower respiratory tract infections in 16 institutions in Japan, and investigated the susceptibilities of isolated bacteria to various anti-bacterial agents and antibiotics and patients' characteristics. Of 499 strains that were isolated from specimen (mainly from sputum) and assumed to be bacteria causing in inflammation, 493 strains were investigated. The breakdown of the isolated bacteria were: Staphylococcus aureus 78, Streptococcus pneumoniae 73, Haemophilus infiuenzae 99, Pseudomonas aeruginosa (non-mucoid) 64, P. aeruginosa (mucoid) 14, Klebsiella pneumoniae 25, Moraxella subgenus Branhamella catarrhalis 21, etc. Of 78 S. aureus strains, those with 4 micrograms/ml or more of MIC of oxacillin (methicillin-resistant S. aureus: MRSA) occupied 53.8%. Vancomycin and arbekacin had the most potent activities against MRSA as observed in 1999. The frequency of S. pneumoniae exhibiting low sensitivity to penicillin (penicillin-intermediate S. pneumoniae: PISP + penicillin-resistant S. pneumoniae: PRSP) was 38.4% being consistent with that in 1999 (34.7%). PRSP accounted for 11.0% of the total, being more than that in 1999 (3.0%). Carbapenems had strong activities against S. pneumoniae. Especially, panipenem inhibited the growth of all 73 strains at 0.125 microgram/ml. Generally, all drugs had strong activities against H. influenzae with MIC80s of 8 micrograms/ml or less. The drug that had the strongest activity against H. infiuenzae was levofloxacin, which inhibited the growth of 94 of the 99 strains at 0.063 microgram/ml. Tobramycin had a strong activity against P. aeruginosa (both mucoid and non-mucoid) with MIC80 of 1 microgram/ml. The mucoid strain was little isolated (14 strains) but the susceptibilities to all drugs were better than the non-mucoid strain. K. pneumoniae showed good susceptibilities to all drugs except ampicillin and the MIC80S were 2 micrograms/ml or less. Particularly, cefpirome, cefozopran, and levofloxacin had strong bactericidal activities against K. pneumoniae with MIC80s of 0.125 microgram/ml, and cefotiam, second-generation cephems, also had a favorable activity being MIC80 of 0.25 microgram/ml. Also, all drugs generally had strong activities against M. (B.) catarrhalis. MIC80s of all drugs were 2 micrograms/ml or less. The drug having the strongest activity was imipenem and levofloxacin inhibiting all 21 strains at 0.063 microgram/ml. Most of the patients with respiratory infection were aged 70 years or older, accounting for approximately a half of the total (44.4%). As for the incidence by the diseases, bacterial pneumonia and chronic bronchitis were the highest, being noted in 38.0% and 31.7% of all the patients, respectively. The bacteria frequently isolated from the patients with bacterial pneumonia were S. aureus (18.3%) and S. pneumoniae (16.1%). In contrast, H. infiuenzae (20.4%) and P. aeruginosa (both mucoid and non-mucoid: 16.7%) were frequently isolated from the patients with chronic bronchitis. Before the drug administration, the bacteria frequently isolated from all the patients were S. pneumoniae (24.3%) and H. infiuenzae (26.7%). The frequency of isolated S. pneumoniae tended to decrease with the increase in the number of administration days while that of isolated H. infiuenzae did not. The frequency of isolated P. aeruginosa tended to increase with the duration of administration. The isolated bacteria were comparable between the patients already treated with penicillins and cephems. In the patients treated with aminoglycosides, macrolides, and quinolones, P. aeruginosa was most frequently isolated (33.3 to 40.0%). PMID- 12532638 TI - [Comparative studies on activities on antimicrobial agents against causative organisms isolated from patients with urinary tract infections (2000). III. Secular changes in susceptibility]. AB - The bacteria (Enterococcus faecalis, Staphylococcus aureus, Escherichia coli, Klebsiella spp. and Pseudomonas aeruginosa) isolated from patients diagnosed as having urinary tract infections (UTIs) in 10 institutions in Japan were supplied between August 2000 and July 2001. Then, the susceptibilities of these bacteria to various antimicrobial agents were examined, and the results were compared with those obtained between 1992 and 1999. Comparison was made by classifying strains isolated from patients into those in uncomplicated UTIs and those in complicated UTIs (including with or without indwelling catheter). E. faecalis showed good susceptibility to ampicillin (ABPC) and imipenem (IPM), and the MIC90s were 2 micrograms/ml. Also, E. faecalis showed good susceptibility to vancomycin (VCM). However, the MIC90, which was 2 micrograms/ml between 1992 and 1999, rose to 4 micrograms/ml in patients with complicated UTIs because the strains inhibited at 4 micrograms/ml increased more than before. The low susceptibility of S. aureus to arbekacin (ABK) in complicated UTIs, as shown in 1998 and 1999, recovered in 2000, and no strains inhibited at > or = 4 micrograms/ml were detected. E. coli showed good susceptibility to CTM (MIC90: 0.25-0.5 microgram/ml) and CZOP (MIC90: < or = 0.125 microgram/ml) and was not resistant to those. E. coli also showed good susceptibility to the other drugs except to penicillins. Decreases in susceptibility of E. coli to quinolones, ciprofloxacin (CPFX), and sparfloxacin (SPFX) were observed in the patients with complicated UTIs. The susceptibility of Klebsiella spp. to all drugs did not significantly change in 2000 and was generally good except to penicillins. Although the susceptibility of P. aeruginosa to carbapenems was notable, the MIC90 went up from 4 micrograms/ml to 16 micrograms/ml in complicated UTIs compared with those observed in the previous year. PMID- 12532640 TI - Catering and special events. Through the holidays and beyond. PMID- 12532641 TI - Care and treatment of residents (TX). Part IV. PMID- 12532639 TI - [Pharmacokinetic and clinical studies on teicoplanin for sepsis by methicillin cephem resistant Staphylococcus aureus in the pediatric and neonate field]. AB - Pharmacokinetics, clinical efficacy and safety of teicoplanin (TEIC) were evaluated in pediatric and neonate patients with MRSA sepsis in the dosages approved in overseas. The administrated dose for pediatrics patients was 10 mg/kg once at hour 0, 12 and 24, followed by every 24 hours intervals. In neonates patients, first dose was 16 mg/kg, then 8 mg/kg every 24 hours intervals. 1. Pharmacokinetic results. All 17 patients (9 neonates and 8 pediatrics) who received TEIC were evaluated for pharmacokinetics. Trough concentrations were analyzed in 16 patients (9 neonates and 7 pediatrics) excluding one patient for lack of measurement of drug concentration at day 7. No patient with a concentration exceeding 60 micrograms/mL in peak or trough concentrations were reported. Mean concentrations in trough at day 3, 4 and 7 in neonates were 15.2, 14.7 and 17.8 micrograms/mL, and in pediatrics were 12.5, 12.2 and 13.1 micrograms/mL, respectively. These results were similar to those reported in foreign pediatrics and neonates patients. 2. Efficacy and safety results. Since no patient was excluded, all patients were evaluated for efficacy and safety. Microbiological efficacy as well as clinical cure were secondarily evaluated in 2 patients for whom MRSA was isolated from blood. Clinical efficacy rate was 76.5% (13/17) and number of cases in judgments of excellent, good, fairly improved and no change were 12, 1, 3 and 1 cases respectively. The patients for whom MRSA was isolated from blood were judged as MRSA eradicated case and cured without any additional anti-MRSA drugs. Adverse events were reported in 2 neonates and 3 pediatric patients. Possibly related adverse events to study drug (adverse drug reactions) were: 1 case of respiratory disorder, thrombocythemia, gamma-GTP increased, GOT increased and GPT increased in 3 pediatrics. These results suggest that an application of overseas dose regimen of TEIC for neonate and pediatrics is appropriate in Japan. PMID- 12532642 TI - Inside the sausage factory: improving estimates of the effects of health insurance expansion proposals. AB - The fate of a proposal to expand health insurance is influenced by predictions of the proposal's effects on the number of newly insured and the cost of new coverage. Estimates vary widely, for reasons that are often hard to discern and evaluate. This article describes and compares the frameworks and parameters used for insurance modeling. It examines conventions and controversies surrounding a series of modeling parameters: how individuals respond to a change in the price of coverage, the extent of participation in a new plan by those already privately insured, firms' behavior, and the value of public versus private coverage. The article also suggests ways of making models more transparent and proposes "reference case" guidelines for modelers so that consumers can compare modeling results. PMID- 12532643 TI - Patents and innovation in cancer therapeutics: lessons from CellPro. AB - This article discusses the interaction between intellectual property and cancer treatment. CellPro developed a stem cell separation technology based on research at the Fred Hutchinson Cancer Center. A patent with broad claims to bone marrow stem cell antibodies had been awarded to Johns Hopkins University and licensed to Baxter Healthcare under the 1980 Bayh-Dole Act to promote commercial use of inventions from federally funded research. CellPro got FDA approval more than two years before Baxter but lost patent infringement litigation. NIH elected not to compel Hopkins to license its patents to CellPro. CellPro went out of business, selling its technology to its competitor. Decisions at both firms and university licensing offices, and policies at the Patent and Trademark Office, NIH, and the courts influenced the outcome. PMID- 12532645 TI - Voices unheard: barriers to expressing dissatisfaction to health plans. AB - Consumers dissatisfied with their health plan can either "exit" (switch service providers) or "voice" (complain to the current provider). Policymakers' efforts to help consumers voice their dissatisfaction to health plans or external mediators have been disappointing, in part because little is known about the determinants of voice. This article represents the first comprehensive assessment of voicing in response to problematic experiences with health plans. A national consumer survey from 1999 is used to test hypotheses about characteristics of problems, patients, and settings that might inhibit voice and assess state regulations intended to enhance voice. Although problems associated with plans led to more voice than exit, voice is circumscribed by several factors: certain groups, such as racial minorities, do not express their grievances as often; episodes with severe health consequences for patients are not reported as regularly. The findings suggest that even though regulatory initiatives have not increased the frequency of voice, they have made grievances more effective, at least in jurisdictions where citizens know about the laws. PMID- 12532646 TI - The foreignness of germs: the persistent association of immigrants and disease in American society. AB - During the 20th century the United States witnessed social, political, and economic transformations as well as advancements in medical diagnosis and care. Despite changes in demography, the meaning of citizenship, and the ability to treat and cure acute and chronic diseases, foreigners were consistently associated with germs and contagion. This article explores why, at critical junctures in American history, immigrants have been stigmatized as the etiology of a variety of physical and societal ills. The article analyzes three periods from 1880 to the present and suggests that now, as germs progressively and, often, indiscriminately cross national, social, and economic boundaries through multiple vectors, the mistakes of the past must not be repeated. Protecting the public health in the current era of globalization requires an ecumenical, pragmatic, and historically informed approach to understanding the links between immigration and disease. PMID- 12532644 TI - Racial and spatial relations as fundamental determinants of health in Detroit. AB - African Americans in the United States have a higher than average risk of morbidity and mortality, despite declining mortality rates for many causes of death for the general population. This article examines race-based residential segregation as a fundamental cause of racial disparities, shaping differences in exposure to, and experiences of, diseases and risk factors. The spatial distribution of racial groups, specifically the residential segregation of African Americans in aging urban areas, contributes to disparities in health by influencing access to economic, social, and physical resources essential to health. Using the Detroit metropolitan area as a case study, this article looks at the influences of the distribution of African American and white residents on access to these resources and discusses the implications for urban policies to reduce racial disparities in health status. PMID- 12532647 TI - [Study of pathogenetic role of Epstein-Barr virus in Hungarian patients with B cell non-Hodgkin lymphomas]. AB - INTRODUCTION: The Epstein-Barr virus is a member of herpesvirus family. It plays an important role in the etiopathogenesis of Burkitt lymphoma, other B-cell non Hodgkin lymphomas, nasopharyngeal carcinoma, X chromosome-linked lymphoproliferative disease, Hodgkin lymphomas and a part of T-cell lymphomas. It has been suggested that association of Epstein-Barr virus with lymphomas and its pathological significance in disease varies in different geographical areas. The aim of our study was to reveal the role of Epstein-Barr virus in B-cell non Hodgkin lymphomas diagnosed in Hungary. The authors asked: A) What is the frequency of the presence of virus genome in the biopsy specimen from B-cell non Hodgkin patients? B) Which types of virus latency can be observed? PATIENTS AND METHODS: Genomic DNA and EBER-specific RNA of Epstein-Barr virus in paraffin embedded specimens were detected by polymerase chain reaction and in situ hybridisation, respectively. RESULTS: Out of 36 B-cell non-Hodgkin lymphoma cases, 16 (45%) were found to be positive for Epstein-Barr virus by the above methods. Expression of latency genes, nuclear antigene-2 and latent membrane protein-1 was studied by immunohistochemical technique. The 16 virus-positive non Hodgkin lymphoma cases showed Epstein-Barr virus latency types II. (37%) or III. (63%). CONCLUSIONS: The authors data suggest that Epstein-Barr virus may be associated with the development of B-cell non-Hodgkin lymphomas in Hungarian patients. PMID- 12532648 TI - [Correlation of lipoprotein(a) and ischemic heart disease]. AB - Increased plasma levels of lipoprotein(a) [Lp(a)] have been considered an independent risk factor for coronary heart disease (CHD). A high concentration of Lp(a) contributes to atherothrombotic risk by multiple mechanisms that include impaired fibrinolysis and increased cholesterol deposition in the arterial wall. Its atherogenicity seems to be related to the similarity in structure with thrombohemostatic factors. The level of Lp(a) in plasma is genetically determined and appears to be unaffected by usual risk factors. There are few medications which have been shown to lower the plasma concentration of Lp(a), however, since these drugs all have multiple other effects their, use is not the choice of the therapy. According to the results of examinations, the best therapeutic option for increased plasma Lp(a) concentration is to lower the plasma level of LDL. PMID- 12532649 TI - [Cerebral uptake and regional distribution of [11C]-vinpocetin after intravenous administration to healthy men: a PET study]. AB - INTRODUCTION: Vinpocetine is a compound widely used in the prevention and treatment of cerebrovascular diseases. The exact mechanism of action of the drug is still not known. The objective of the present investigation was to determine the global uptake and regional distribution of radiolabelled vinpocetine in the human brain. Three healthy persons were examined with positron emission tomography (PET) and [11C]-vinpocetine. RESULTS: The uptake of [11C]-vinpocetine in brain was rapid and on average as a maximum 3.7% of the total radioactivity injected was in the brain 2 minutes after radioligand administration. The uptake was heterogeneously distributed among brain regions. When compared with the cerebellum, an a priori reference region, the highest regional uptake was in the thalamus, the upper brain stem, the striatum and the cortex. CONCLUSIONS: The brain regions showing increased uptake in the human brain correspond to those in which vinpocetine has previously been shown to induce elevated metabolism and blood flow by PET clinical studies in patients with chronic ischaemic post-stroke condition. PMID- 12532650 TI - [Determination of serum semicarbazide-sensitive amine oxidase activity in diabetic retinopathy in type-2 diabetes]. AB - INTRODUCTION: Recent data suggest that the copper-containing semicarbazide sensitive amine oxidase enzyme (SSAO) may play a role in vascular endothelial damage through conversion of certain endogenous monoamines, like methylamine into cytotoxic aldehydes, hydrogen peroxide and ammonia. SSAO is present in various human tissues and in the serum. Elevated SSAO activities have been reported in patients with both types of diabetes mellitus. The purpose of this study was to examine the possible association between serum SSAO activity and the different severity stages of diabetic retinopathy. PATIENTS AND METHODS: A prospective study was performed on a defined group of Type 2 diabetic patients (n = 93) compared to non-diabetic control subjects (n = 42). All participants underwent a detailed ocular examination (slit lamp, colour retinal photography, fluorescein angiography) and standard laboratory investigations. Age at diagnosis, duration of diabetes, presence of systemic hypertension, medication and BMI were recorded. Serum SSAO activity was determined by a radiometric procedure using [14C] benzylamine as substrate. RESULTS: In the total group of Type 2 diabetic patients SSAO activity (mean +/- SD) was significantly elevated compared to non-diabetic controls (n = 93, 131.72 +/- 53.07 vs. n = 42, 89.56 +/- 26.89 pmol.mg-1 protein.hour-1, p < 0.0001). After dividing patients to four subgroups according to the severity of diabetic retinopathy, serum SSAO activity was significantly higher in patients with high-risk proliferative diabetic retinopathy (n = 16, 166.96 +/- 70.56 pmol.mg-1 protein.hour-1) compared to those without retinopathy (n = 42, 119.54 +/- 50.49 pmol.mg-1 protein.hour-1, p < 0.02). CONCLUSION: The results support the hypothesis that elevated SSAO activity may be involved in the pathogenesis of microvascular diabetic late complications, such as retinopathy. The pharmacological manipulation of SSAO activity might be an interesting new concept for prevention and treatment of diabetic retinopathy. PMID- 12532651 TI - [Endometrial cancer in a young woman taking oral contraceptives for a long time]. AB - The authors report a rare case of endometrial adenocarcinoma occurred in a 35 year old woman, who had taken oral contraceptives for 13 years. She came for consultation with irregular vaginal bleeding. Diagnostic curettage and histologic examination confirmed the diagnosis as endometrial adenocarcinoma. This case deserve special attention due to the rarity of the disease among young women. The authors emphasize the necessity of appropriate diagnostic work-up. In such cases histological examination is strongly recommended, since irregular vaginal bleeding might be caused by malignant disease even at young women using sequential oral contraceptives. PMID- 12532652 TI - [Remarks about anthrax]. PMID- 12532653 TI - [Cases of "incidental" syphilis]. PMID- 12532654 TI - [Role of focal infections]. PMID- 12532655 TI - Postmenopausal hormone therapy: have HERS2 and WHI given us any new information? AB - There still remains no consensus on the role of HRT in the initiation of breast cancer. Stimulation of an otherwise occult breast cancer leading to more frequently diagnosed breast cancers may seem more feasible and supported by the survival data. Although HERS and WHI both fail to offer support for a cardiovascular protective effect of HRT, other studies differ in their conclusions. There is still no true primary prevention randomized trial, and we may never have one. To take or not to take HRT may be one of the most important decisions a woman makes in her lifetime. For millions of women, now expected to live into their ninth decade, this is more than a theoretical question; it is a potentially life-altering one. For the majority of women, the cancer-phobia that leads to an avoidance of HRT is neither rational nor based on scientific evidence. However, there may be a relatively small subset of women for whom HRT does pose a potential health risk. Every health decision a woman makes includes such an assessment of risk balanced against benefit. It is the health provider's obligation to participate in risk assessment and assist each woman in designing a plan for life-long good health. The well-informed and consenting woman (and this is more important now than ever before) will make the correct decision for her circumstances. From a public health perspective, it is clear that more woman years are saved as a result of the beneficial effects of HRT than are lost from its negative effects. PMID- 12532656 TI - Foot problems and foot care practices in diabetes. A survey of public and private diabetes clinics affiliated with a university hospital. PMID- 12532657 TI - Participation in physical activity to lower the risk of heart disease or stroke. PMID- 12532659 TI - What is a doctor? Reflections on the physician charter of professionalism. PMID- 12532658 TI - Successful treatment of obsessive compulsive disorder with oxcarbazepine. A case report. PMID- 12532660 TI - Dr. Bartolo M. Barone: a remembrance. PMID- 12532661 TI - [The malignant side of chronic inflammatory bowel disease]. AB - Patients with chronic inflammatory bowel disease (IBD) have a higher risk of colorectal carcinoma, and present with this malignancy at a younger age than non IBD individuals. In three patients, two men aged 20 and 36 years and one woman aged 34 years, colorectal cancer developed at a young age, following a long history of ulcerative colitis. Surveillance for colorectal cancer in IBD patients needs to be performed by regular colonoscopy with extensive biopsy sampling for the detection of dysplasia, regarded by many as predictive for colon carcinoma. Whenever a dysplasia-associated lesion or mass or a highgrade dysplasia is identified, there is a strong indication for colectomy. When low-grade dysplasia is found, the findings should be discussed with the patient and it should be decided whether the patient should resume surveillance or opt for colectomy. PMID- 12532662 TI - [Nobel Prize for Physiology or Medicine 2002 is awarded for research into the genetic regulation of organ development and programmed cell death]. AB - The human body consists of hundreds of cell types, all originating from one fertilized egg. During the embryonic and foetal periods, the number of each cell type increases dramatically. The cells mature and become specialised to form the various tissues and organs of the body. Large numbers of cells are also formed in the adult body. Cell death is a normal process, which runs parallel to this generation of new cells. It takes place in both the foetus and adult, thereby maintaining the appropriate number of cells in the tissues. This delicate, controlled elimination of cells is termed programmed cell death or apoptosis. This year's Nobel laureates in Physiology or Medicine (Sydney Brenner, H. Robert Horvitz and John E.Sulston) have made seminal discoveries relating to the genetic regulation of organ development and programmed cell death. The nematode Caenorhabditis elegans was used as an experimental model system for following cell division and differentiation from the fertilized egg to the adult. The laureates have identified key genes that regulate organ development and programmed cell death and have also shown that corresponding genes exist in higher species, including humans. The discoveries are important for medical research and have shed new light on the pathogenesis of many diseases. PMID- 12532663 TI - [The return of sleeping sickness in an epidemic form: international action for drugs]. AB - In Central Africa, the number of people suffering and dying from sleeping sickness is the same as it was in 1920, whereas the disease was under control in the 1950s. In Zaire (Congo), Gambiense sleeping sickness was virtually under control by 1960. Due to war, chaos, population movements, lack of resources, and collapse of public health, the situation is now as it was 80 years ago and some years ago it was even more bleak as the supply of drugs seemed to halt. Due especially to the action of Medecins sans Frontieres, the availability of the four essential drugs pentamidine, suramine, melarsoprol and eflornithine is now secure until 2006. Control of the epidemic in Central Africa can be achieved once peace and order and a basic infrastructure have been restored, and under the condition that adequate resources will be available. PMID- 12532664 TI - [Immunomodulating strategies in the treatment of sepsis]. AB - Sepsis is associated with the activation of several inflammatory cascades, including the cytokine network and the coagulation system, but it can also be associated with an immunodepressed state. This can lead to a situation in which the septic patient becomes more susceptible to secondary infections. In animal experiments, inhibition of the cytokine cascade by administration of tumour necrosis factor alpha (TNF alpha)-receptors or anti-TNF alpha antibodies has led to reduced mortality, but this has not been confirmed in clinical trials. After the data were pooled, there was a statistically significant decrease in mortality of 3-5%. Treatment with endotoxin antibodies, corticosteroids in high doses, other anti-inflammatory agents and agents designed to eliminate immunodepression generally also did not produce a convincing decrease in mortality. Research on antithrombotic agents has yielded, along with disappointing results with antithrombin III and 'tissue factor pathway inhibitor', one study with a positive result. In septic patients with organ failure who were treated with activated protein C, a coagulation inhibitor, the mortality decreased from 30.8 to 24.7%. PMID- 12532665 TI - [Treatment and prevention of haemorrhage with recombinant activated factor VII, not only in hemophilia]. AB - Based on the improved insight into the mechanism of blood coagulation in vivo, recombinant factor VIIa (rFVIIa) has been developed as a pro-haemostatic agent. It has been shown that this treatment is effective in patients with haemophilia and an inhibiting antibody against a coagulation factor. Administration of rFVIIa can be effective in the treatment of bleeding episodes and the prevention of perioperative blood loss in a number of other coagulation disorders as well. Initial clinical studies indicate that administration of rFVIIa may also reduce blood loss in patients with severe bleeding without pre-existent abnormalities in blood coagulation, but this requires confirmation in additional clinical trials. PMID- 12532666 TI - [Diagnostic image (119). A neonate with abnormal distention and vomiting. Meconium ileus and cystic fibrosis]. AB - A female neonate developed abdominal distension with vomiting. She was suffering from meconium ileus and cystic fibrosis. PMID- 12532667 TI - [From gene to disease; 'frame shift'-mutation in the CARD15-gene and Crohn's disease]. AB - A pericentromeric region on chromosome 16 (IBD1 locus) has been linked with Crohn's disease (CD). Very recently, three genetic variants in the CARD15 gene within the IBD1 locus have been identified which were highly associated with CD. Carriage increases the relative risk of developing CD. One specific mutation (3020insC) leads to a stop codon and truncation of the C-terminal tandem leucine rich repeats (LRR) of the CARD15 protein. Of all patients with Crohn's disease, 11-19% are heterozygous and 3-7% homozygous for this frameshift mutation. The CARD15 gene is expressed in monocytes. The LRR-domain is thought to be involved in the binding of bacterial lipopolysaccharide and subsequent activation of nuclear factor kappa-B (NF kappa B). NF kappa B plays a central role in the regulation of the expression of other genes involved in the inflammatory response. In vitro, embryonic kidney cells transfected with the CARD15 3020insC mutant showed a reduced activity of NF kappa B after exposure to lipopolysaccharide compared to cells transfected with the wild-type CARD15 gene. How the reduced response to lipopolysaccharide contributes to CD is not yet clear. PMID- 12532668 TI - [Favourable effect of a standard elimination diet on the behavior of young children with attention deficit hyperactivity disorder (ADHD): a pilot study]. AB - OBJECTIVE: To determine whether a standard elimination diet can decrease the ADHD symptoms in a heterogeneous group of young children with ADHD. DESIGN: Open, descriptive. METHOD: 40 children, 36 boys and 4 girls, aged 3-7 (average 4.8 years), who met the DSM-IV-criteria for ADHD, followed their usual diet for two weeks and thereafter for two weeks an elimination diet, based on the few foods diet (rice, turkey, pear and lettuce). The behaviour of the child was evaluated at study entry, after the baseline period and at the end of the diet. Parents completed the 10-item Conners list, the ADHD Rating Scale and a physical complaints list. The teachers completed the 10-item Conners list and the ADHD Rating Scale twice, at the beginning and at the end of the diet. RESULTS: According to the parent-ratings, 25 children (62%) showed an improvement in behaviour of at least 50% on both the Conners list and the ADHD Rating Scale at the end of the elimination diet. Nine children (23%) withdrew from the study because the parents were unable to stick to the diet or because the child fell ill. Among the 15 children with both parent and teacher ratings, 10 responded both at home and in school. CONCLUSION: In young children with ADHD an elimination diet can lead to a statistically significant decrease in symptoms. PMID- 12532669 TI - [Rising costs of drugs in hospitals in the period 1996-2000 and over the next few years]. AB - OBJECTIVE: To obtain an impression of drug expenditure in hospitals and in particular the costs due to novel more expensive drugs both in the past and in the future. DESIGN: Descriptive. METHOD: Data on intramurally supplied drugs were collected from 6 of the 95 general hospitals, 5 of the 14 top clinical hospitals, 4 of the 8 university hospitals and I of the 13 categorical hospitals for the period 1 January 1996-31 December 2000. The data were extrapolated to the entire of the Netherlands per hospital category and per year on the basis of the adherence figures. The drug costs were calculated on the basis of cost prices. For the most important new potential drugs it was ascertained whether they would actually become available and if they would substantially contribute to the drug expenditure. RESULTS: In 2000, the total drug expenditure within all Dutch hospitals was estimated to be [symbol: see text] 402 million. A substantial part of the costs could be attributed to the use of anticancer drugs (19%) and antibiotics (14%). In 2000, about 12% of the drug expenditure could be attributed to the use of novel, expensive drugs. The total costs of drugs increased on average by 8% per year during the period 1996-2000. The contribution of novel, expensive drugs nearly doubled during this period, and anticancer drugs were the most significant factor in this. A large proportion of the potentially new drugs were innovative anticancer drugs and drugs used to treat immune diseases. These compounds are likely to be expensive. Based on these findings it is expected that over the next few years, drug expenditure within hospitals to grow by at least 20% per year. PMID- 12532670 TI - [Three patients with African sleeping sickness following a visit to Tanzania]. AB - Three Dutch tourists, a man aged 57 and two women aged 55 en 52 years, acquired African trypanosomiasis in the national parks of Tanzania. Two, without central nervous system involvement, were cured after treatment in the Netherlands, albeit one after having suffered a relapse. In the third patient, involvement of the central nervous system was diagnosed in Africa and she was treated with melarsoprol. After an apparently uneventful recovery she was readmitted with cerebral complaints and symptoms. While being treated with melarsoprol she lapsed into coma. She died following repatriation. An epidemic of trypanosomiasis is currently raging through Central Africa. In several western countries, trypanosomiasis has been diagnosed recently in tourists who visited Tanzania. PMID- 12532671 TI - [Parenteral administration of low dose ketamine for the treatment of neuropathic pain in cancer patients]. AB - In three patients, a 52-year-old woman with skeletal metastases from bladder carcinoma, a 54-year-old man with metastasised thyroid carcinoma and a 40-year old man with a non-Hodgkin lymphoma, neuropathic pain developed that could not be alleviated adequately by patient-controlled opioid administration. It is known that ketamine, a N-methyl-D-aspartate (NMDA) receptor antagonist, can improve opioid-induced analgesia. Pending invasive therapy, the three patients were given a continuous low dose of parenteral ketamine (2-5 micrograms/kg/min). The pain in the first two patients responded so well to ketamine that they decided to waive the invasive pain treatment and to continue the ketamine infusion at home until death. In the third patient, the addition of ketamine resulted in an adequate level of analgesia during the waiting period for invasive treatment with an intrathecal catheter. PMID- 12532672 TI - [Increased perioperative blood loss during treatment with paroxetine]. PMID- 12532673 TI - [Better clinical effects for carpal tunnel syndrome with surgery than with splinting: a randomized clinical trial]. PMID- 12532674 TI - [Better clinical effects for carpal tunnel syndrome with surgery than with splinting: a randomized clinical trial]. PMID- 12532675 TI - [Finger prints on human skin--results of a general applications-oriented series]. AB - Numerous methods have been described for making dactyloscopic traces visible on human skin, but these have never become routine--last but not least because they are too complicated and time-consuming to perform. The purpose of the present investigation was to find out to what extent fingerprints on the skin of deceased persons can be made visible with the help of conventional methods like iodine fuming or brushing with Magna-Brush and can be preserved with gelatine film or moulding material. Of the total number of 486 experimental fingerprints the dermal ridges could be made visible in 150 prints. The best findings were obtained when the trace had been applied several hours post mortem. On the skin of the extremities better results could be achieved than on the trunk or the neck. Dactyloscopic tracing on the skin of bodies in homicide cases should be taken into consideration especially if there is evidence that the perpetrator touched the victim after his/her death (e.g. if the body was transported). PMID- 12532676 TI - [Discovery, assessment and reconstruction of homicide with photographs of the murder]. AB - Report on a case in which photographs of the murder were the first hint pointing to homicide. Due to the advanced decomposition of the body the photographs were essential for the reconstruction of the homicide, in particular for the differentiation of the intravital injuries against postmortem lesions and the chronological order of the acts of violence. PMID- 12532677 TI - [Serum norharman and harman analysis using high pressure liquid chromatography/mass spectrometry and value of beta-carbolines as blood alcohol markers]. AB - Firstly, a method for LC-MS/MS-analysis of the beta-carbolines norharman and harman in serum was established and validated. Secondly, serum samples from persons during ethanol loading conditions were investigated (n = 26). Norharman was regularly found positive only in persons with BAC > 1.6@1000. In this subgroup harman was detected in 5 out of 9 cases. The finding of norharman concentrations > 50 pg/mL in the serum of 4 out of 5 control persons was of high interest. In addition serum samples of smokers (n = 9) were analyzed for the beta carbolines. All samples from smokers were tested positive for norharman with concentrations > 50 pg/mL in 7 cases. These results were in accordance with recent reports in literature and underline that the beta-carbolines norharman and harman do not meet the criteria of alcohol state-markers and positive serum sample testing may result from endogenous as well as from exogenous sources. PMID- 12532678 TI - [Hemopericardium caused by endocarditis ulcerosa: an unusual cause of sudden death in adolescence]. AB - The authors report on the case of a 18-year-old student who died suddenly and unexpectedly of a pericardial tamponade after perforated ulcer-polypous endocarditis of the aortic valve. The detection of aerococcus urinae--which usually causes urinary tract infection--on the inflamed aortic valve demonstrates an unusual pathogenetic chain linking complexes of findings in the urogenital system and the heart. PMID- 12532679 TI - [Bioterrorism--also a challenge for forensic medicine?]. AB - In view of current events the question is discussed to what extent the risk of bioterrorism may be an issue relevant for forensic medicine also in Germany, although at present there seems to be no concrete threat. The cases which became known so far were either false alarms or foolish pranks (copycats), which have to be, and are indeed, prosecuted by the state (section 126 German Criminal Code). Reference is also made to the measures of disinfection recommended by the Robert Koch Institute. PMID- 12532680 TI - [Practical application of the Injury Severity Score (ISS) in expert forensic testimony]. AB - The retrospective analysis of the autopsy records of 50 homicides showed that the Injury Severity Score (ISS), a numerical scoring system initially developed to quantify the severity of injuries sustained in road traffic accidents, can also be useful for objectively describing and ranking the overall severity of trauma with regard to forensic issues. The present case report illustrates to what extent the ISS can help to assess the contribution of each assailant in homicides committed by several perpetrators. In the case presented the court was convinced that one perpetrator had inflicted four deep stab wounds to the victim's face (each with bony lesions), 2 stabs to the chest piercing the right lower pulmonary lobe and causing a haemothorax of 200 ml, an abdominal stab wound without involvement of a parenchymatous organ as well as multiple defence wounds of the arms. Thereafter, a second perpetrator was thought to have inflicted several heavy blows with a full water bottle causing severe contusions on the right side of the forehead, the chin, the left side of the face and a spider's web fracture of the frontal bone. Using the ISS an injury severity score of 24 was assigned to the first complex of injuries and a score of 10 to the second complex. The forensic conclusions with regard to prognosis and lethal outcome are discussed. PMID- 12532681 TI - Priority-setting ethics in public health. AB - Unlike the well-developed system of ethical priority-setting that physicians and bioethicists have developed in the field of organ transplantation, public health lacks a standardized and publicly recognized system for making ethical resource allocation decisions. Such a system would be useful for the purpose of countering prejudice-based arguments that tend to hinder the progress of public health programs aimed at marginalized groups. By examining organ transplantation priority-setting criteria as they relate to public health issues, this article seeks to stimulate debate about the varieties of criteria that should and should not be used in public health decision-making. PMID- 12532682 TI - State medical marijuana laws: understanding the laws and their limitations. AB - Significant attention has been given to the debate regarding allowances for medical marijuana use since the 1996 California and Arizona ballot initiatives. State medical marijuana allowances, however, have existed since the mid-1970s. Much of the current debate stems from confusion about the various ways states approach the issue. In this paper, we present original legal research on current state medical marijuana laws identifying four different ways states statutorily enable the medical use of marijuana. We discuss the tension these approaches have with federal law as well as their implications regarding real access for patients. In addition, we present information on how a small number of states are trying to deal with the issue of access within the context of their medical marijuana laws, and discuss the implication of various supply approaches on the enforcement of other state marijuana laws. PMID- 12532683 TI - Knocking at freedom's door: race, equity and affirmative action in U.S. higher education. PMID- 12532684 TI - Chernobyl and iodine deficiency in the Russian Federation: an environmental disaster leading to a public health opportunity. AB - The Chernobyl nuclear disaster of April 26, 1986, triggered a chain of devastating events that later included an unexpected increase in childhood thyroid cancer and evidence of iodine deficiency (ID) in Russia. For the Russian people the Chernobyl event had profound psychological impacts, provoking anxiety about nuclear technology and mistrust of governmental control efforts. Frequently in public health a crisis is required to create the political will to manage longstanding problems, and public health officials must rapidly mobilize to take advantage of the opportunity. In this case, ID, previously not seen as a problem in Russia, was recognized to be potentially serious, and the Russian Federation, assisted by the catalytic bi-national effort of the U.S.-Russian Joint Commission on Economic and Technological Cooperation (Gore-Chernomyrdin Commission (GCC)) established a model salt iodization policy, developed a planning process, and implemented a program to prevent ID through a systematic approach that included the people, government, and private groups using open communication, dissemination of the findings, and action plans. By 1999, political will had been mobilized and over 20% of the nation's salt was being iodized, up from about 1% in 1996. Universal iodization of salt was not a specific objective of the GCC; however, the increasing availability of iodized salt is leading to the elimination of ID, which is now a political goal in Russia. The full realization of this goal will require more time for education, marketing, and possibly legislative action. PMID- 12532685 TI - Fighting for America's energy independence. PMID- 12532686 TI - Fred Soper and the global malaria eradication programme. PMID- 12532687 TI - [Advanced age and risk factors for acute myocardial infarction]. AB - This case-control study, analyzed the role of coronary risk factors in acute myocardial infarction (AMI) in the elderly, and established the nature of this association and the degree of risk. Data were derived from an investigation (1060 cases and 1071 controls) conducted in 35 coronary care units from clinical centres in Argentina between November 1991 and August 1994. Our analysis was based on data collected from subjects over age 65. Cases were 427 patients with AMI and without history of ischaemic heart disease. Controls were 396 subjects identified in the same centres as the cases. Odds ratios (OR) estimates and their 95% confidence intervals (CI) were derived from multiple logistic regression equations including terms for age, education, social status, smoking status, history of diabetes or hypertension, body mass index and family history of coronary heart disease. The risk factors independently and strongly related to the risk of AMI were the following: hyperlipidemia (serum cholesterol > or = 240 mg/dl): OR = 1.76 (95% CI: 1.25-2.49), smoking habits: OR = 1.6 (95% CI: 1.06 2.4), hypertension: OR = 2.05 (95% CI: 1.51-2.73), diabetes OR = 1.71 (95% CI: 1.12-2.70), one relative with family history of coronary heart disease: OR = 1.36 (95% CI: 0.93-1.97) and two or more relatives: OR = 2.63 (95% CI: 1.21-5.71). This study confirms in the elderly the importance of hyperlipidemia, tobacco, hypertension, diabetes and family history of coronary heart disease as risks factors of AMI. PMID- 12532688 TI - [Pulse wave velocity and urinary albumin excretion in hypertensive patients treated with perindopril]. AB - Systolic and diastolic blood pressures and urinary albumin excretion (UAE) have been recognized as predictors for cardiovascular risk. Furthermore, arterial compliance (AC) disorders assessed by increased aortic pulse wave velocity (PWV) are closely related to changes in blood pressure and strongly correlated with cardiovascular mortality and presence or extent of atherosclerosis. Our purpose in the present study was to determine a relationship between AC using PWV and UAE in a group of non-smoking patients with essential hypertension, and the level of interaction of ACE inhibition on these two variables. A total of 70 non-smoking never treated hypertensive patients (33 men and 37 women), aged 50 +/- 7 years (range 35-69), have been enrolled in this study. All of them underwent PWV by a computerized device (Complior) and UAE determination by radial immunodiffusion method, on baseline and after six months of treatment with perindopril (4.6 +/- 1.4 mg/day). We have found a significant decrease of systolic blood pressure (160.2 +/- 10.6 vs. 131.9 +/- 7.1 mmHg, p < 0.01), diastolic blood pressure (100.6 +/- 5 vs. 81.6 +/- 4.8 mmHg, p < 0.01), PWV (13.4 +/- 1 vs. 9.1 +/- 0.9 m/sec, p < 0.01), and UAE (42.2 +/- 19.3 vs. 11.1 +/- 3.6 mg/day, p < 0.01) at the end of the sixth month when they were compared to baseline values. Furthermore, renal function was also improved by the treatment at the end of the study as illustrated by creatinine clearance (87.5 + 22.5 vs. 102.1 + 23.5 ml/min, p < 0.01). Moreover, a high positive correlation between UAE and PWV at the beginning of the study (r = 0.81; p < 0.01) and after six months of treatment (r = 0.66; p < 0.01) was observed. In addition, PWV vs. UAE, differences between sixth month and baseline have shown a high correlation (r = 0.67; p < 0.01) and using a multiple regression test we found that PWV (t ratio 5.76; p < 0.001) was the most important and significant independent variable that correlates with UAE. These results suggest the existence of a real link between UAE and AC in non smoking patients with arterial hypertension, and that ACE inhibition can similarly modify these two parameters. PMID- 12532689 TI - Effect of the dose of oral hydrocortisone on growth rate during long-term treatment of children with salt losing congenital adrenal hyperplasia. AB - The effect of the dose of oral hydrocortisone on stature growth rate of patients with the salt losing form of congenital adrenal hyperplasia and adequate electrolyte balance was here assessed. Thirty patients (21 girls and 9 boys) were followed longitudinally for 0.52 to 8.64 years, between chronological ages 0.35 and 8.64 years. Nine consecutive periods (Ps) of follow up were defined in order to compare two auxological parameters, height (H) at the end of a follow up P and delta H standard deviation score (SDS). According to growth rate during a particular P, two types of Ps were defined: Ps with delta H SDS > -0.5 (Group 1, satisfactory growth rate) and Ps with delta H SDS = or < -0.5 (Group 2, poor growth rate). A cut off value of 18.5 mg/m2/day of oral hydrocortisone (95% CI upper limit of group 1) was defined to separate acceptable from excessive doses. In P2, mean (+/- SD) H SDS (-1.81 +/- 1.15) was significantly lower than in any of the other Ps (p < 0.001). In P1 and in P2, delta H SDS was negative, but it was positive in P3 and in P4. Hydrocortisone dose in P1 and in P2 was significantly higher than in the rest of the Ps. All patients in P1 and most patients in P2, but not in other Ps, received excessive doses. Predicted adult H, calculated in 9 patients was not statistically different from their respective target H. It is concluded that, during the first year of life, our patients received an excess of oral hydrocortisone (> 18 mg/m2/day) and grew poorly, but they were able to recover, at least temporarily, when the dose was adjusted during the following years. PMID- 12532690 TI - [Evolutionary and bone marrow recovery indicators in bone marrow transplantation after total body irradiation]. AB - Oxidative stress and reticulocyte maturity index (RMI) were studied in 27 patients who underwent bone marrow transplantation (BMT). Plasmatic lipoperoxide levels of those patients with unfavorable evolution were significantly increased on days 12-14 post-transplant (median 1.83 microM, range 0.78-5.82) compared with preconditioning levels (median 1.05 microM, range 0.36-1.84) (p < 0.05). Patients with favorable evolution revealed significantly higher lipoperoxide levels during conditioning regime (median 1.42 microM, range 0.31-4.50) (p < 0.05). Starting from the 3rd post-transplant week a significant and continuous decrease was observed, with a median of 0.77 microM (range 0.21-1.48 p < 0.05) for the 3rd, and a median of 0.60 microM (range 0.11-1.48 for the 4th week (p < 0.01). A significant increase in total antioxidant activity was observed in the three patients who died up to the 35 days post-transplant. Recovery of bone marrow function was detected by RMI after a median time of 17 days (range 11-24) post allogeneic transplantation. The threshold established for absolute neutrophil count was achieved after a median of 21 days (range 14-28) (p < 0.001). An increase of plasma lipoperoxides on days 12-14 post-transplant may be a predictive value of unfavourable evolution. RMI was the earlier indicator of engraftment in allogeneic BMT. PMID- 12532691 TI - [Tegumental leishmaniasis in Las Lomitas, Province of Formosa, Argentina, 1992 2001]. AB - Las Lomitas Hospital reported 85 cases of tegumentary leishmaniasis between 1992 and 2001. The cases were males (72.9%), 91.8% older than 10 years, 10.5% presented mucosae involvement. A single ulcer on lower limbs was the most frequent manifestation. The epidemiological and clinical patterns were consistent with the infection due to Leishmania (Viannia) braziliensis. The annual distribution of ulcers onset was bimodal, the frequency of cases had a significant association with the Bermejo river volume, and the rainfall of the previous year. Intradermoreaction and recalling questions suggested that fishing in the river was the main risk activity. Among 1018 Phlebotominae collected, 97.1% were Lutzomyia neivai, 2.3% L. migonei and 0.6% L. cortelezzii (0.6%), the peridomestic/gallery forest of the Bermejo river proportion was 1/9, with similar composition to the populations around the same river in Salta province. The increase of cases since 1992, the ongoing environmental changes due to developmental works, and the potential installation of a periurban transmission cycle, require an active surveillance of cases and entomological components in the Chaco region. PMID- 12532692 TI - [Incidental adrenal histoplasmosis]. AB - An adrenal mass found in the course of abdominal ecographic or computed tomography (CT) techniques performed for other reasons is a new diagnostic challenge. The patient in this case was asymptomatic and without previous illness. An ecographic study revealed an enlarged right adrenal gland, 66 by 33 mm. CT showed bilateral irregular enlargement of both adrenal glands. Nonadrenal malignancies and hyperfunctioning tumors were ruled out. A CT-guided biopsy was performed, and the histopathologic study was consistent with the diagnosis of histoplasmosis. The patient lived in an area endemic for this disease, and had daily contact with bird droppings. Histoplasmosis is a rare unsuspected cause of bilateral adrenal masses in the asymptomatic immunocompetent host. PMID- 12532693 TI - [Cytomegalovirus cellulitis]. AB - Cutaneous lesions in CMV infection are rare, often a late manifestation of systemic infection, and usually herald a fatal course. A 70 year-old woman received a kidney transplantation one month before consulting and immunosuppressive therapy that included cyclosporine A and methylprednisone. She complained of fever, local pain in her right leg, and an erythematous and swelling plaque. She was treated with intravenous antibiotics without improvement. A skin biopsy was performed and the tissue obtained was sent for bacterial and fungal cultures as well as for histological examination. Cultures were negative. The biopsy showed CMV cytopathic changes. Immunoperoxidase staining was positive for CMV and polymerase chain reaction (PCR) testing revealed CMV DNA. She was treated with ganciclovir with resolution of the lesion. CMV cellulitis is a rare cutaneous manifestation which has not been previously reported. PMID- 12532694 TI - [Pure red cell aplasia after allogeneic transplantation of ABO incompatible hematopoietic stem cells]. AB - ABO incompatibility in allogeneic bone marrow transplantation may be associated with incomplete or delayed erythroid engraftment, being pure red cell aplasia (PRCA) the most severe complication in this setting. Attempts for the treatment of PRCA have been made with erythropoietin or with plasmapheresis with relative success, and some authors have reported the reversibility of PRCA with antilymphocyte globulin (ALG or ATG), based on the assumption that PRCA might be immunologically mediated. We report herewith a patient with acute leukemia who developed post--BMT pure red cell aplasia. His sibling donor (sister) was HLA identical and ABO incompatible, having low agglutinin titers against donor's blood group. PRCA did not improve after treatment with erythropoietin or a boost of hematopoietic progenitor cells obtained from donor's peripheral blood but the problem was resolved completely after treatment with ALG. PMID- 12532695 TI - [Pelvic tumor and bilateral hydronephrosis]. PMID- 12532696 TI - [Definitive pacemaker, acute infarct and reperfusion]. PMID- 12532697 TI - [Endogenous endophthalmitis due to candida]. PMID- 12532698 TI - [Childhood tuberculosis and its diagnosis in Argentina]. AB - TB notification rate in infants and children 0-4 years was 24.3/100,000 in Argentina, in 2000, for a global incidence of 31.8/100,000. A 17% of all TB cases occurred in children (0-14 years), indicating a very active TB transmission. Usually TB is diagnosed when the disease is already advanced. Main criteria for the diagnosis are: a previous contact with an infectious TB patient, a positive PPD test result and clinical--radiological signs and symptoms. Bacteriology plays a limited role in diagnosis; gastric aspirate is the most often used specimen for bacteriological examination in children. Culture is performed on solid egg-media, but other more recently developed culture media systems are available in reference laboratories. Nucleic acid amplification assays are also used, but its systematic quality control is still lacking. Their results should be considered only as complementary. Strengthening of the laboratory network will contribute to increase diagnostic certainty of childhood TB, to improve reliability of cases reported, and to a better knowledge of the drug resistance profile. Early diagnosis of TB in children, by contacts investigation of infectious patients, followed by preventive chemotherapy or treatment when necessary, may revert the current epidemiological situation. TB meningitis in the 0-4 years group showed a significantly more pronounced decline than that of adults TB for the period 1980 2000, (only 10 meningitis cases were diagnosed in year 2000, 0.29/100,000), confirming the protection conferred by BCG vaccination when applied to newborns with a continuously high coverage. PMID- 12532699 TI - [Telomeres and genomic damage repair. Their implication in human pathology]. AB - Telomeres, functional complexes that protect eukaryotic chromosome ends, participate in the regulation of cell proliferation and could play a role in the stabilization of genomic regions in response to genotoxic stress. Their significance in human pathology becomes evident in several diseases sharing genomic instability as a common trait, in which alterations of the telomere metabolism have been demonstrated. Many of them are also associated with hypersensitivity to ionizing radiation and cancer susceptibility. Besides the specific proteins belonging to the telomeric complex, other proteins involved in the DNA repair machinery, such as ATM, BRCA1, BRCA2, PARP/tankyrase system, DNA PK and RAD50-MRE11-NBS1 complexes, are closely related with the telomere. This suggests that the telomere sequesters DNA repair proteins for its own structure maintenance, which could also be released toward damaged sites in the genomic DNA. This communication describes essential aspects of telomere structure and function and their links with homologous recombination, non-homologous end joining (NHEJ), V(D)J system and mismatch-repair (MMR). Several pathological conditions exhibiting alterations in some of these mechanisms are also considered. The cell response to ionizing radiation and its relationship with the telomeric metabolism is particularly taken into account as a model for studying genotoxicity. PMID- 12532700 TI - [Angiogenesis in cancer]. PMID- 12532701 TI - [Organ transplantation. What is the limit?]. PMID- 12532702 TI - [Certification of physicians in the specialty of Clinical Medicine]. PMID- 12532703 TI - [Stenosis of the external auditory canal: etiology, classification and management]. AB - Stenosis of the external auditory canal is uncommon, poly-etiological pathology of the external auditory canal. The management of stenosis of the external auditory canal is difficult because of re-stenosis and danger of postoperative complications. The purpose of this article is to present modern classification, etiopathogenesis of the external auditory canal stenosis, indications for the surgical management, some aspects of the surgical management, using the modern literature and clinical cases. PMID- 12532704 TI - [Importance of biologically active components and plants in the prevention of complications of diabetes mellitus]. AB - Diabetes complications, especially late (chronic) ones, are the main reasons of invalidity and early mortality. The most threatening diabetes complications are vascular and metabolic complications (diabetic neuropathy, angiopathy, cataract, glaucoma, optic neuropathy, retinopathy, diabetic nephropathy). Good diabetes control is very important, because in early stages these changes are reversible. In order to decrease the number of diabetes complications and to postpone their development, the use of biologic active components and plants is recommended. The most important biologic active substances for this purpose are vitamins and minerals, proteins, polysaccharides, lectins, saponins and flavonoids. According the scientific data, the mostly used plants are: Ginkgo biloba, Allium sativum, Silybum marianum, Panax Ginseng, Carica papaya, Vaccinium myrtillus, Phaseolus vulgaris. Some of them are proposed for treatment of symptoms related to venous and lymphatic vessel insufficiency, for the prophylaxis and treatment of liver damage caused by metabolic toxins, in chronic degenerative liver conditions, for the therapy of digestive disorders, to increase in the unspecific way the resistance of the organism to various environmental influences, and to stabilize membranes through antioxidant and radical scavenging actions. PMID- 12532705 TI - [Spontaneous subarachnoid hemorrhage: patients' examination after aneurysm negative initial angiograms]. AB - AIM OF THE STUDY: To find out, which patients in case of subarachnoid hemorrhage and negative initial angiograms need a repeated investigation, when it should be repeated and what kind of angiographic investigation should be performed. MATERIALS AND METHODS: Fifty seventh patients with subarachnoid hemorrhage and negative initial angiograms were investigated in Department of Neurosurgery of Kaunas University of Medicine Hospital from January 2001 to August 2002. They underwent a repeated angiography (digital subtraction or computed tomography angiography) generally in the period of 10 days. RESULTS: Repeated cerebral angiography revealed saccular aneurysms of various locations in 23 patients. Perimesencephalic hemorrhage was present in 24 cases and no aneurysms were detected in these cases. Aneurysm was suspected in initial angiography in 14 cases and 10 of them were confirmed after second angiography. In all cases of application computed tomography angiography provided equal or more information for neurosurgeon compared to digital subtraction angiography. Nearly half of the detected aneurysms (n = 11) were small (< 5 mm). CONCLUSIONS: Repeated angiography is not recommended in case of perimesencephalic pattern of spontaneous subarachnoid hemorrhage. In case of suspicion of aneurysm in initial angiography, computed tomography angiography could be a method of choice performed immediately. PMID- 12532706 TI - [Phonatory characteristics following different endolaryngeal microsurgical techniques in Reinke's disease]. AB - The aim of the study was to compare different surgical techniques in treatment of Reinke's edema according to subjective and quantitative changes of voice characteristics. We evaluated 62 surgically treated patients from 34 to 68 years (mean 47.7 years) with Reinke's edema. Two endolaryngeal microsurgical methods were used: classical as mechanical stripping of the mucosa-Ist, mechanical stripping group (n = 30), and removal of fluid through aspiration with preservation of vocal cord medial edges mucosa-IInd, preserving mucosa group (n = 32). Clinical voice assessment was composed of laryngoscopy, subjective voice evaluation (GRBAS hoarseness scale, visual analogue scale of voice quality, and pitch) and quantitative voice evaluation (seven quantitative voice parameters obtained from voice range profile (VRP) and registering of maximum phonation time). Patients were evaluated twice--before operation and at mean 23.8 days after. Postoperative follow-up data was registered from 57 patients: 28 from Ist and 29 from IInd groups. According to our data the positive dynamics for all voice parameters was observed in both patients' groups. Surgical approach with preservation of vocal cord medial edges mucosa on overall benefit after operation according to the voice quality is statistically significantly better compared to classical operation technique (chi 2 = 8.9, p = 0.029). While comparing these two approaches statistically significant differences were found for subjective voice parameters--grades of hoarseness and roughness, and for quantitative VRP parameters: pitch range, maximum-minimum intensity range, total voice area, and overall vocal dysfunction degree. The mean values of most voice parameters in early postsurgery period significantly differed from normal. Clinical voice assessment may be useful for evaluation of surgical treatment efficacy as well as for comparison of surgical techniques. PMID- 12532707 TI - [Quantitative assessment of acute myocardial infarction size. Impact of early mechanical recanalization of infarct related artery]. AB - The aim of investigation: 1) to determine the diagnostic value of QRS score and LV echocardiography in assessment of the size of myocardial infarction in acute stage, 2) to establish the impact of infarct related artery recanalization on myocardial infarction size. METHODS AND MATERIAL: In order to investigate whether infarct size could be estimated by QRS scoring system soon after reperfusion we evaluated QRS score obtained serially before and twice after reperfusion, and the echocardiographic global EF in 57 patients with acute myocardial infarction who underwent successful mechanical recanalization of infarct related artery. Coronary flow in infarct related artery was evaluated by the Thrombolysis in Myocardial Infarction trial (TIMI) criteria. QRS scores were calculated according to the method of Wagner (37 criteria and 29 points). The electrocardiographic ejection fractions (EFECG) were calculated according to Roubin method, and the global echocardiographic ejection fractions (EFECHO) were calculated according to Simpson method. All patients were divided into 3 groups according to the quality of myocardial reperfusion expressed as intensity in change of electrocardiographic phases at the time of infarct related artery recanalization (1 gr.--the change of ECG phases > or = 2; 2 gr.--the change of ECG phases through 1; and 3 gr.--with no changes of ECG stages). All patients had an effective infarct-related artery recanalization expressed as 2 or 3 TIMI grade. RESULTS: A low and insignificant correlation was observed between EFECG and EFECHO for patients with anterior myocardial infarction (r = 0.35) and for patients with posterior myocardial infarction (r = 0.12). The EFECG had a tendency to be lower in patients with worse myocardial reperfusion (from 59.06 +/ 6.12 in 1 gr. to 50.93 +/- 10.87 in 3 gr). At this time the EFECHO was almost the same in all groups of patients. Additionally, the EFECHO was significantly lower than EFEKG in all groups of patients (p = 0.000017-0.001). The QRS score had general tendency to increase after infarct-related artery recanalization, however the most evident increase was obtained in 1 gr. of patients with rapid change of ECG stages. A significant correlation (r = 0.87 for patients with anterior myocardial infarction and r = 0.85 for patients with posterior myocardial infarction) was observed between QRS scores obtained after infarct related artery recanalization and that obtained after 10-12 days. In conclusion, QRS score and EFEKG better than EFECHO reflects the myocardial infarction size in acute stage. Increasing of the myocardial infarction size after infarct-related artery recanalization is connected with reperfusional injury more expressed in patients with more effective myocardial reperfusion. PMID- 12532708 TI - [Effect of blood and valve cultures on complication rate and outcome of infective endocarditis (analysis of data of patients treated at Kaunas district hospitals, 1999-2001)]. AB - The objective of the study was to determine the current epidemiology of infective endocarditis (IE) and to evaluate correlation of blood and valve cultures with complication rates of the disease. During 1999-2001 138 patients, 91 (65.9%) males and 47 (34.1%) females with mean age of 50 +/- 16, were observed. The incidence of IE makes 0.0042% per year in Kaunas district, in-hospital mortality rate was 24.6%. Forty five patients had positive blood culture, 54.8%--negative blood cultures, 16.9% of valve tissue cultures were positive. Patients with positive blood culture had higher incidence of acute IE, vascular phenomena, peripheral abscess and toxic encephalopathy. Culture negative endocarditis presented itself with worse left ventricular function, more frequent immunologic phenomena, myocarditis, glomerulonephritis, cardiac operation and perivalvular leak. Positive valve culture was associated with more frequent perivalvular leak. Mortality rate did not differ in groups. PMID- 12532709 TI - [Efficiency of rehabilitation methods in the treatment of arm lymphedema after breast cancer surgery]. AB - In this article we analyze influence of rehabilitation methods in treatment of arm lymphedema. In Kaunas oncological hospital were examined 60 women after surgery for breast cancer. The work objective was to evaluate efficiency of rehabilitation methods in treatment of arm lymphedema and in evaluate movement amplitude of shoulder joint. Two groups of women depending on rehabilitation start were evaluated. The same methods of rehabilitation were applied to both groups: physical therapy, electrostimulation, massage, lymphodrainage with apparate. Our study indicated that women, who were treated at early period of rehabilitation (3 months), showed statistically significantly (p < 0.01) better results in increase of movement amplitude of shoulder joint. However, results of treatment of arm lymphedema, comparing with women who started rehabilitation after 12 months, were equally successful--results were not statistically significantly better (p > 0.05). PMID- 12532710 TI - [Development and analysis of revitalizing tincture]. AB - The article deals with the production of the complex tincture of Hyperici herba 80.0 g, Cinnamomi cortex 20.0 g, Rosmarini folium 20.0 g, Lavandulae flos 20.0 g, Zingiberis rhizoma 60.0 g and the selection of an extracting liquid phase, and the choice of extraction method. The experimentally determined optimal conditions for tincture preparation were as follows: concentration of ethanol--70% (V/V), the particle size of the leaves, herb, flowers--2.0-3.0 mm, for the bark and the roots--0.3-0.5 mm, the extraction method--percolation. The quality of the tincture was evaluated by analyzing the sum of flavonoids (recalculated into equivalent amount of rutin), dry residue, the concentration of ethanol, density of the extractive preparation, and microbial contamination. Also the overall appearance was checked, the identification of flavonoids was performed by color reactions and high-pressure liquid chromatography. PMID- 12532711 TI - [Analysis of psychotropic drug mixtures using high-pressure liquid chromatography in acute poisoning cases]. AB - The qualitative and quantitative method of determination of amitryptilin, codeine and fluoxetine in the mixture using high-pressure liquid chromatography is described in this paper. Chromatogram is presented which shows, that preparations are fully separated and do not interfere each others analysis. Tables with chromatographical separation characteristics are also presented. Proposed calibration curves of quantitative analysis and calculated medium relative error of quantitative ascertainment for every preparation of the mixture are shown. Final conclusion: method is applicable for qualitative and quantitative analysis of amitryptiline, fluoxetine and codeine in the mixture in hasty poisoning cases. PMID- 12532712 TI - [Malignant tumor incidence in employees of the Alytus textile factory (1978 1997)]. AB - The purpose of this study was to evaluate cancer incidence in the large cotton manufacturing factory in Lithuania. Altogether 10,198 workers employed at least 1 year in 1969-1997 were included in the cohort and followed during the period 1978/01/01-1997/12/31. National cancer rates were used to calculate the expected number of cancer cases. The overall cancer risk for men was slightly higher than that in the general population (standardized incidence rate (SIR) 1.15, 95% confidence interval (95% CI) 0.98-1.34). A significant increase in the incidence of esophagus (11 observed cases, SIR 3.76, 95% CI 1.88-6.67) and slightly increased of lung (42 observed cases, SIR 1.26, 95% CI 0.91-1.70) cancer became evident. None of the cancer risk showed statistically significant excess cancer incidence in the textile-processing (spinning and weaving) departments (SIR 0.98). In the women cohort the level of the general incidence was very close to expected, standardized incidence rates (SIR) being 0.99 (95% CI 0.88-1.13). However, there was a significant increase in the number of cases of gall bladder (6 observed, SIR 3.19, 95% CI 1.17-6.95). The analysis of the results among textile-processing (spinning and weaving departments) workers indicated the elevated risk of breast cancer (44 observed cases, SIR 1.49, 95% CI 1.08-2.0) and cervical cancer (24 observed cases, SIR 1.68, 95% CI 1.08-2.50). The number of lung cancer cases in this group was a higher, but statistically not significant (5 observed cases, SIR 1.53, 95% CI 0.5-3.58). Increased SIR values were observed for > or = 10 years since the first exposure for all cancers, cervix uteri, ovary and kidney. CONCLUSIONS: The overall cancer risk for men cohort was slightly higher than that in the general population. There was a significant increase in the number of cancer of the esophagus. The overall excess risk in women cohort was only for gall bladder, but for spinners and weavers the elevated risk was for cervix uteri and breast cancer. After 10 years of employment the excess risk was already for all cancers, cervix uteri, ovary and kidney malignant tumors. PMID- 12532713 TI - [Efficiency of postoperative sanatorium rehabilitation of patients with hip joint endoprosthesis: importance of kinesitherapy]. AB - When treating degeneration diseases, endoprosthesis operations are performed widely. The results of the treatment depend not only on the operating technique and up-to-date prostheses but also on the correct postoperative rehabilitation, applied in particular the kinesitherapeutic means. The aim of the research was to establish the effect of postoperative rehabilitation with wide use of kinesitherapeutic means in sanatorium conditions on the functional capacity of the joint operated on as well as on the general condition of the patient. The research has been done at the traumatology department of the Sanatorium "Versme" in Birstonas. The patients were prescribed a complex of kinesitherapeutic means, i.e. curative gymnastic exercises performed for 45 min twice a day, water kinesitherapy, self-dependent isometric exercises, walking and manual massage. Clinical examination of 30 patients has been undertaken on the first and last day of their rehabilitation in the sanatorium. The mobility of the hip joint, muscle strength and circumference of the thigh have been measured. The patient's tolerance to physical load, his/her sensitiveness to pain, the patient's rate of walking and self-care have been estimated. It has been established that kinesitherapeutic means applied during the 30 day course of rehabilitation undergone in the Sanatorium increased the amplitude (flexion and drawing back) of the joint operated on, the strength of quadriceps muscle femoris and the strength of drawing back and back femoral muscles, as well as decreased sensitiveness to pain and improved the rate of walking and functional class (p < 0.001). The circumference of the femoral muscle and functional independence of the patients remained unchanged. The functional capacity of the endoprothesized hip joint of the patients improved to some extent irrespective of the section performed during the operation as well as irrespective of the sex and age of the patients. PMID- 12532714 TI - [Prof. Alfonsas Kaikaris -- the founder of the Lithuanian Pharmacy Museum: his personality and scientific activities]. AB - Alfonsas Kaikaris (1922-1997) was a professor of pharmacy at the Kaunas Medical Institute (now University), historian, museologist, founder of the Lithuanian Pharmacy Museum and of the field of pharmacy history in Lithuania. A. Kaikaris was born in Zagare in 1922. He received his pharmacy diploma in 1947 and began work at the university. Following the reorganization of the university into two institutes, he went to work in the Medical Institute, where he served as the vice dean of the Pharmacy and Stomatology Department from 1957 until 1963. He began to collect pharmaceutical artifacts in 1957. In 1973, the Institute provided a small room in the attic for these objects and so a small pharmacy museum, the first in Lithuania, was born. Thanks to the hard work of A. Kaikaris and others, today this museum has grown into the Pharmacy and Medical History Center of Lithuania, whose work is widely known throughout Europe. A. Kaikaris was also a member of the board of the Lithuanian Scientific Society and head of the Society's Pharmaceutical History Section from 1964 until he retired in 1987. He is the author of 67 scholarly papers and numerous popular articles and conference presentations. In 1988, he received the Paul Stradins Award from the Paul Stradins Medical History Museum in Riga, Latvia, and until his death in 1997 worked as a consultant to the Lithuanian Museum of the History of Medicine and Pharmacy. PMID- 12532715 TI - [The 100th anniversary of the birth of the phthisiatrist Jonas Runkevicius]. AB - In 2002 phthisiologist J. Runkevicius turned 100 years of age. Associate professor doctor J. Runkevicius (1902-1966) was the first in midwar Lithuania to commence epidemiological investigations of tuberculosis and continued them all his life. Having graduated from the Vytautas Magnus university, from 1933 to 1937 he worked as a physician at the Kaunas Red Cross tuberculosis sanatorium and in 1937-1940 as a head physician at dr. K. Grinius sanatorium of the Society for the fight against tuberculosis. In 1938-1939 he improved his skills at the tuberculosis treating institutions in Switzerland and Finland. In 1940-1941 and 1944-1945. J. Runkevicius was a head physician of the Alytus national tuberculosis hospital and in 1944-1945--chief of the health care department in the Alytus district, too. After his studies J. Runkevicius began his scientific researches. The object of his research was teachers of primary schools, their infectiosity with tuberculosis, and the dependence of morbidity with tuberculosis on the living conditions. Dr. J. Runkevicius generalized his researchers in his thesis "Tuberculosis in bourgeois Lithuania according to clinical roentgenelogical examination data of consecutive groups", maintained at the Faculty of Medicine of Vilnius university in 1949. Since 1945 up till his death in 1966, Dr. J. Runkevicius has been working at the Faculty of Medicine of Vilnius University, chair of therapy. In 1945-1946 concurrently he headed the Department of Tuberculosis of the Ministry of Health. In 1947-1949 he was a deputy dean of the Faculty of Medicine. While being a lecturer of Vilnius university, Dr. J. Runkevicius also worked as head of the organizational-methodic department of the National scientific research institute of tuberculosis. Doc. Dr. J. Runkevicius was one of the first phthisiotherapists in Lithuania who made every effort in the organization of preventive roentgenelogical examination of teachers and students with the view of diagnosing the cases of tuberculosis. Dr. J. Runkevicius supervised the scientific research work of physicians, he was written more than 30 papers. While working at the National scientific research institute of tuberculosis and Vilnius university, Dr. J. Runkevicius has done a lot in the fight against tuberculosis in Lithuania. PMID- 12532716 TI - [Concomitant chemo-radiation treatment of malignant tumors: biological bases]. AB - Concomitant radio-chemotherapy is widely used in the treatment of various localisations of cancer. Numerous interactions are known between ionizing radiation and cytotoxic agents, which in vitro may modify the dose-response relationships. The main interactions are spatial cooperation, independent cell kill, cell synchronization, inhibition of repair of radiation damage, reoxygenation, reduction of the hypoxic fraction and increased apoptosis. Exposure of irradiated cells to cytotoxic agent lead to increase the number of unrepaired double-strand breaks. By this mechanism the action of DNA repair of both therapeutic modalities enhance therapeutic efficiency. The main effects of chemoradiotherapeutic combination in the laboratory are inhibition or antagonism, sub-additive, additive and supra-additive effects. The most important for clinical practice are supra-additive effects, when the cytotoxic effect of the combination is greater than the sum of the effects of radiotherapy alone and that of chemotherapy alone. It is however difficult to extrapolate laboratory and experimental results to clinical practice because the conditions for interaction of chemo- and radiotherapy in vitro are different. Clinical conditions usually are more compatible for additive and sub-additive interactions. The main role of laboratory and experimental investigations is to propose new drug--radiotherapy combination to phase II trials. PMID- 12532717 TI - [Schizoaffective disorder: clinical symptoms and present-day approach to treatment]. AB - During 20th century serious mental disorders were divided into two groups according symptomatology and course of disorder. Individuals with dominating disturbance of perception, thinking and cognition were basically diagnosed having schizophrenic. Individuals with mood disturbance were basically diagnosed having affective disorders. However, there were patients who did not fit neatly into either category. In 1933 Jocob Kasanin introduced the term "schizoaffective psychosis". Scientific discussions involved the possibility that schizoaffective disorder was conceptualized most accurately as following: a type of schizophrenia, a type of affective disorder, a unique disorder that was separate from both schizophrenia and bipolar disorder, an arbitrary categorization of clinical symptoms that marked a continuum between schizophrenia and affective illness, a heterogeneous collection of "interforms" between schizophrenia and affective disorders. However, diagnosis of schizoaffective disorder is included both in DSM-IV-TR and ICD-10. Schizoaffective disorder is listed in the category "schizophrenia and other psychotic disorders". The differential diagnosis includes basically either schizophrenia or affective disorder. The epidemiological status of schizoaffective disorder is somewhat uncertain compared with schizophrenia because of dilemmas related to diagnosis and classification of the disorder. Treatment of schizoaffective disorder comprises psychotropic medication, supportive psychotherapy, social care, rehabilitation. The most important groups of psychotropic medications are: antipsychotics, antidepressants and mood stabilizers. Atypical antipsychotics are the first-line medication for schizoaffective disorder due to their pharmacological properties. In the case of schizoaffective disorders combination of atypical antipsychotics with antidepressants seems to be useful. Novel antidepressants have priority for the combination mentioned above. Peculiarities of mechanism of action of antidepressant are important for combinations. Mood stabilizers seem to be useful for treatment of certain type of schizoaffective disorder as well. PMID- 12532718 TI - [Middle ear cholesteatoma: present-day concepts of etiology and pathogenesis]. AB - Since J. Cruveilhier described cholesteatoma as the "pearly" tumor of the middle ear in 1828, the pathogenesis of cholesteatoma remained controversial. It is accepted that cholesteatoma may be congenital or acquired. Several pathogenic mechanisms have been proposed to explain the pathogenesis of congenital cholesteatoma. Proposed theories include ectopic epidermis rest, ingrowth of meatal epidermis, metaplasia and reflux of amniotic fluid. Four basic theories present the pathogenesis of acquired cholesteatoma: invagination of the tympanic membrane (retraction pocket cholesteatoma), basal cell proliferation, epithelial in-growth through a perforation (the immigration theory) and squamous metaplasia of middle ear epithelium. The aim of the article is to review the recent literature dealing with problems of the etiopathogenesis and classification of cholesteatoma. PMID- 12532719 TI - [Impact on environmental factors on the reproductive system and fetal development]. AB - A literature review discusses the effect of selected environmental factors on women reproductive system, fetal development and growth. According to recent reports, 2-3% of newborns have congenital malformations. These malformations are caused by interaction of genetic and environmental factors. Exposure of paternal or maternal organisms to environmental hazards may damage germ cells or interfere fetal development, resulting in malformation of various organ systems. Since environmental hazards exposures are complex, it is difficult to establish the primary effect of single factor. Factors, that are known to increase the risk of congenital malformations, preterm delivery or spontaneous abortion, are classified into five groups--psychological, social, biological, physical and chemical factors. The governments of most counties recognize the effect of hazardous environmental factors on public health as global problem. World Health Organization encourages researches, aimed at evaluation of various environmental factors impact on health of pregnant women and their offsprings. PMID- 12532720 TI - [Patient-controlled analgesia in the treatment of postoperative pain in children and adolescents]. AB - OBJECTIVE: We aimed to evaluate the efficacy, morphine requirements and side effects in patients managed with patient-controlled analgesia postoperatively. METHODS: Eighty-seven patients, 7 to 22 (median 14) years of age, after orthopedic, abdominal or thoracic surgery, were studied retrospectively. All patients had a preset standard continuous morphine infusion of 10 micrograms/kg/h, 20 micrograms/kg bolus dose, 8 min lock-out time and 100 micrograms/kg/h maximal dose. All patients were additionally treated with non steroidal anti-inflammatory drugs. Respiratory rate, pain intensity, morphine requirements and nausea/vomiting were evaluated every 3 hours. Pain was assessed using 6-point scale (1-no pain, 6-the worst pain). RESULTS: Median time for pain management was 51 h. Median (range) used dose of morphine was 19.9 (4.9-75.2) micrograms/kg/h. Sixty-one percent of the patients had a mean pain score of 1 or 2, 26%--3, 13%--4 or 5. Nausea/vomiting occurred in 78% of cases. Respiratory rate less than 10 breaths per min was observed in 5.7% of patients. No patient required opioid antagonist to treat respiratory depression. CONCLUSIONS: In the majority of patients patient-controlled analgesia was effective and used doses of morphine were low. Though nausea/vomiting was the most common side effect, decreased respiratory rate was observed in some patients. Regular patient evaluation for vital signs and side effects is recommended. PMID- 12532722 TI - [Early diagnosis of spinal cord schwannoma: the significance of the pain syndrome]. AB - In this study we present 76 cases of spinal cord schwannoma, treated in the Clinic of Neurosurgery Kaunas University of Medicine Hospital. The analysis of early clinical symptoms in diagnosis of schwannoma and the value of the pain syndrome as the first sign in manifestation of illness is emphasized. The results of operated patients in early stage are presented as well. The first symptom of schwannoma is pain; in 81.6 percent of cases it is radicular and in 11.8 percent- local. Early stage (radiculopathy) we determined to 19.7 percent of patients. Majority (95.8 percent) of operated patients in early stage improved and recovered. Neurologists and doctors of general practice must pay great attention to complaints of patients and course of illness when there are no results after 2 months of conservative treatment, they must sent the patient to neurosurgeon. PMID- 12532721 TI - [JTc prolongation in hypertrophic cardiomyopathy amd arterial hypertension]. AB - Pathological long duration of QTc and JTc intervals is non-questionable factor of sudden death. Information of the investigation of electrocardiogram increases during physical exercise test or transesophageal pacing of the left atrium. OBJECTIVE: The search of the factors of sudden death in hypertrophic cardiomyopathy and arterial hypertension patients and healthy persons from their electrocardiographic changes of JTc interval applying transesophageal pacing of the left atrium. MATERIAL AND METHODS: It was done transesophageal pacing to 12 healthy persons, 12 arterial hypertension and 13 hypertrophic cardiomyopathy patients. Pacing rates of 80, 100 and 130 bpm were applied for 3 min each. RESULTS: There was no prolongation of JTc in control group. JTc prolongation (more than 0.35 s) appeared in 3 arterial hypertension patients and 3 hypertrophic cardiomyopathy patients. Five hypertrophic cardiomyopathy patients and one arterial hypertension patient had prolonged JTc interval at the rest electrocardiogram. CONCLUSIONS: Transesophageal pacing of the left atrium helped to detect pathological long JTc interval in arterial hypertension and hypertrophic cardiomyopathy patients, when JTc interval was not prolonged at the rest electrocardiogram. PMID- 12532723 TI - [Prognosis of the surgical treatment of parasagittal meningioma]. AB - The neurosurgical treatment results of parasagittal meningiomas were analyzed in the following article. It retrospectively reviewed 132 patients with parasagittal meningiomas operated in the Department of Neurosurgery during the period of 1995 2000. Forty-four (33.3%) meningiomas invaded the superior sigittal sinus (SSS) and partially or totally obturated it. One hundred-five (79.5%) meningiomas were removed totally and post-surgery results of 111 (84.1%) patients were good. On discharge 17 (12.9%) were in poor condition, and 4 (3%) patients died. Postoperative complications occurred in 46 (34.8%) patients. Venous infarction was the most frequent complication in this study. It developed in 12 (9.2%) patients. The outcome after removal of parasagittal meningiomas was related to the patient condition prior to surgery (p = 0.000), postoperative complications (p = 0.001), tumor site (p = 0.05) and invasion of SSS by the tumor (p = 0.04). Venous infarction was the most frequent postoperative complication. It is possible to reach total meningioma's removal by ablation of SSS and replacing it by peripherical venous. PMID- 12532725 TI - [Evaluation of ultrastructural changes in the ischemic myocardium in surgically obtained biopsies of the left ventricle]. AB - Electron microscopic evaluation of the degree of ischemic injury in diseased human hearts is a very useful tool in estimating the quality of any method of myocardial preservation. Ultrastructural alterations give accurate data in the clinical setting. Our semiquantitative scoring system was created considering existing evaluation systems and based on our own observations. Eight grades of ischemic injury were defined, ranging from 1 (normal ultrastructural appearance) to 8 (irreversible ischemic damage) with respect to integrity and swelling of mitochondria, nuclei, myofilaments, membranes, organelles and components, as well as extracellular structures. PMID- 12532724 TI - [Treatment of multiple lacerated wounds of the face and upper limbs]. AB - We describe a case of severe face and upper extremities injury obtained in accident. 30 cm x 12 cm left arm soft tissue defect covered with thoracodorsal myocutaneous flap after necrectomy. After 10 days a microvascular transplantation of scapular fasciocutaneous flap was performed thus covering a 15.8 cm x 10.5 cm soft tissue defect in face. Eighteen days after trauma autodermoplastica was done in a 35 cm x 11 cm would of the right arm. The patient was discharged after 6 weeks of treatment. October 19, 2000 reconstruction of left ulnar nerve (n. ulnaris) was performed using sural nerve (n. suralis) grafts. Debulking of scapular flap in face was done simultaneously. Eighth months after trauma the patient returned to work. A good esthetic effect was observed. PMID- 12532726 TI - [Development of "Saules" tincture technology]. AB - The article deals with the production of the "Saules" tincture in a way of selecting an extractive agent concentration, the size of particles of the raw material and the method of extraction. The experiment indicates that the optimum concentration of ethanol in the tincture is 70% V/V, the particles' size is in the range from 1 to 2 mm; the production method recommended for use is percolation. The quality of the tincture was evaluated by determining the quantity of flavonoids and dry residue, the concentration of the ethanol, the density and the microbial contamination. The presence of quercetin, rutin and hyperoside was confirmed using high-pressure liquid chromatography. The stability of the tincture was evaluated. PMID- 12532727 TI - [Development of technology of the enzyme preparation: procelan ointment]. AB - When investigating the release of procelan from diphilic, hydrophobic and hydrophilic basis we determined that the best release of the active substance is from hydrophilic basis. The macrogolic base when compared with methylcellulosic one shows clearly expressed osmotic properties and therefore is more suitable as a base for ointments intended to treat suppurative wounds. This was confirmed by treatment of experimentally excited suppurative wounds on rabbits. Hydrophilic ointments containing lyophilized procelan powders from 4 per cent to 20 per cent were examined. The most remarkable therapeutic effect reached with the ointment of macrogolic base, optimal concentration of procelan being 10 per cent. The method of assay of procelan in the ointment was created. The created dosage form is stable for three years at room temperature. PMID- 12532728 TI - [Analysis of flavonoids in the flowers and leaves of Monarda didyma L]. AB - Monarda didyma L. is a herbaceous perennial with the square stems and opposite leaves characteristic of herbs in the Labiate family. Monarda didyma L. is often used as a domestic medicine, being particularly useful in the treatment of digestive disorders. The leaves and flowering stems are antihelmintic, carminative, diuretic, expectorant, febrifuge, rubefacient and stimulant. Monarda didyma L. has twenty-six compounds in the essential oil, of which thymol, with smaller amounts of para-cymene, d-limonene, carvacrol, linalool, and hydrothymoquinone. A new method was used for the determining of the amount of separate flavonols by high-performance liquid chromatography. The flavonoids (rutin, hyperoside, quercitrin, luteolin, quercetin) were found in leaves and flowers of beebalm. The quantity of flavonoids was higher in flowers than in leaves of Monarda didyma L. PMID- 12532729 TI - [Survival after myocardial infarction among the middle-aged Kaunas men and women]. AB - The aim of the present study was to determine and compare the short-term and long term survival after myocardial infarction (MI) in the Kaunas men and women aged 25-64 during 1983 to 1998, according to ischemic heart disease register data. The source of the data--Kaunas population-based ischemic heart disease register. The diagnosis of MI was based on the criteria defined by the WHO MONICA Project. All myocardial infarction events that occurred among Kaunas population aged 25-64 years during 1983-1998 were recorded. Myocardial infarction survival was evaluated using Kaplan-Meier method and using z test for comparing the survival curves. RESULTS: The data of 3,613 persons with a first myocardial infarction and 528 with a recurrent myocardial infarction were analyzed. Detected, that among men and women a short-term (28 days) survival probability was significant different. In the men aged 25-64 short-term survival after a first myocardial infarction was better comparing with the women survival (z = 4.63, p = 0.03). The short-term survival among men aged 25-64 with a recurrent myocardial infarction comparing with woman did not differ. The long-term survival probability in patients, who survived the first 28 days, among men and women with a first and recurrent myocardial infarction aged 25-64 was without statistically significant differences. PMID- 12532730 TI - [Opinion of Lithuanian population about accessibility and quality of primary health care services]. AB - OBJECTIVE OF THE STUDY: To assess the opinion of Lithuanian adult population about accessibility and quality of primary health care services. MATERIAL AND METHODS: The study was conducted in 2001. A national random sample of 5000 Lithuanians aged 18 years and older has been taken out of the National Population Register. The questionnaires were filled in by 3514 persons (response rate- 71.6%). RESULTS: Sixty-six percent of respondents reported that they have received health care at an outpatient health facility in the last 12 months. Women visited a doctor more frequently than men did; older inhabitants more frequently than younger ones. Fifty-eight percent of respondents reported they had no problem to get a health care provider they were happy with, 16.5% had a mild problem and 10.1%--severe or extreme problem. Fifty-four percent of respondents noted that they had no problem to use other health services than they usually went to, while 15.6% reported--it was a severe or extreme problem. The majority of respondents (59.5%) answered that they always got care as soon as they wanted. Patient opinion about quality of primary health care services depended on how well doctors, nurses or other health care providers communicated with them, i.e. whether patients were treated with respect and dignity, whether they were listened carefully, whether they were explained things understandably etc. Seventy-three percent of respondents reported that health care providers communicated with them well or very well. Older respondents assessed the communication with doctors better than younger did. CONCLUSION: The data of regular questioning of population could be used for the successful health care reform implementation. PMID- 12532731 TI - [Amniotic fluid embolism]. AB - Amniotic fluid embolism is a rare obstetric emergency in which amniotic fluid, fetal cells, hair, or other debris enters the maternal circulation. Amniotic fluid embolism is an incompletely understood obstetric complication unique to pregnancy presenting with the acute onset of hypoxia, hypotension and severe coagulopathy. Up to 86% of the cases are fatal, maternal death occurs within the first hour of symptom onset in 50% of the patients. It cannot be predicted nor prevented. The epidemiology of amniotic fluid embolism, frequency, pathophysiology, clinical presentation, histologic findings, diagnosis, differential diagnosis, possible treatment, mortality rate, neonatal survival are discussed in this review article. PMID- 12532732 TI - Integrative physiology and organ systems research and the reorganization of study sections at CSR. PMID- 12532733 TI - Ignore lies about animal research. PMID- 12532734 TI - [Effect of pantogam on visual function and hemodynamic of eyes in patients with primary open-angle glaucoma]. AB - The complex therapy in patients with primary open-angle glaucoma, with normalized intraocular pressure but with declined visual functions included the use of the Russian neurometabolic preparation pantogam. The improvement of the visual function appeared as enlargement of the borders of visual fields, decreased area and sensitivity deficit (according to computer perimetric data). Doppler ultrasonography has demonstrated an increase in the mean blood velocity in the orbital vessels and a decrease in eyeball blood supply defects after pantogam therapy which is a favourable sign of a glaucomatous process. The authors consider the use of pantogam to be rational in the complex therapy of patients with glaucoma. PMID- 12532735 TI - [Modern notions of regularities and mechanisms of formation of Frank-Kamenetskiy glaucoma]. AB - Frank-Kamentsky glaucoma, a hereditary disease of the eyes, transmitted by female partners with X-linked type. It is accompanied by typical progressive lesions in the stroma and in the iridal pigment sheet at the background of goniodisgenesis. Males suffer from these diseases most frequently. Glaucoma develops in the 2-nd- 3-rd decades of life. Its course resembles that of in course open-angle glaucoma. Megalocornea and goniodysgenesis are aggravating factors pre-disposing to glaucoma. Microsymptoms in the iris of female conductors can serve as a prognostic test in planning progeny. PMID- 12532736 TI - [Laser instrument surgery in preventing and treating retinal detachment (communication II)]. AB - The paper is devoted to the actual problem of ophthalmology--removal of cysts and retinal detachments caused by cystoid retinal lesions. 3 stages of treatment are identified: stage 1 used YAG-laser dissection of the external wall of the retinal cyst with drainage of its contents. Stage 2 involved extrascleral filling reinforced by argon coagulation of the zone of the emptied cyst or by endolaser coagulation. Laser intervention was performed by the laser combine "Visulas-YAG II" produced by the firm "Karl Zeiss" (Germany) and fitted up by endolaser output. Coagulation energy was 0.2-0.7 W, the power of YAG-laser was 0.1-5.7 mJ with impulses of 120. Almost in all of the cases laser action was realized during one session. The laser's action was associated with hemorrhage in 11 cases (57.9%), and within 1 year of observations 2 relapses were noted (10.5%). YAG perforation of the external wall of a retinal cyst brings on its drainage and evacuation that is a perspective factor in subsequent surgical interventions during operations for retinal detachment due to cystoid degeneration. PMID- 12532737 TI - [Features of treating patients with combined damage to the orbit and eye in the late period in peacetime and war conditions]. AB - Analysing the diagnosis and treatment of 100 patients with combined injuries of the organ of vision and orbital zone of the face provided clinical features of lesions in peace-time or battle accidents. Battle accidents were found to be prognostically worse for visual functions. The use of immunotherapy for inflammatory complications of a traumatic process is demonstrated to be advisable. A program of stepwise treatment of combined lesions of the eye and the orbit with the priority of preserving the function of the vision organ. At stage 1 it is necessary to take measures for restoration of visual functions to avoid their decline or loss. At stage 2 reconstructive interventions in the orbit and visceral cranium, directed to the provision of topographic and trophic conditions for further restoration of eye functions should be performed. One-stage surgical intervention may be used if there are no real danger of vision loss. The peculiarities of reconstructive treatment of multifractional fractures of the orbit in the battle accident and fragmental fractures of the orbit in peace-time accident are described. Patients with combined injuries of the vision organ and the orbit in specialized institutions should be cared. PMID- 12532738 TI - [Postoperative complications of penetrating subtotal keratoplasty in children]. AB - Accordingly to experience of corneal transplantations during 1981-2001 in the children's clinic of the Helmholz Moscow Research Institute of Ocular Diseases the authors present the clinical picture, rate, prognostic value and modes of correction of postoperative complications after keratoplasty. A retrospective study analyzed the incidence of complications after 94 penetrating keratoplasties performed in 82 children aged from 10 months to 14 years in 89 eyes for congenital corneal opacity (n = 22) or corneal opacity caused by trauma (n = 40) or inflammation (n = 20). 60 (73%) within 1 year after operation developed different reactions and complications due to transplant failure in 41 (44%) cases. Among them there were graft rejection episodes (34%), anterior synechiae and retrocorneal membrane formation (17%), donor corneal neovascularization (11%), elevated intraocular pressure (11%). There were a few cases with shallow anterior chamber, hyphema/hemophthalmos, retinal detachment, primary graft failure, corneal ulcer, suture infiltrates and abscesses. Subatrophy of the eye was an outcome in 3 cases. The development of complications after penetrating keratoplasty is determined mainly by the physiology of children, age, severity of underlying and concomitant pathology of the eye. Nevertheless, the odds of donor tissue for transparent graft survival increase if such complications are detected and removed. PMID- 12532739 TI - [Original method of extracapsule fragmentation of the lens nucleus during phacoemulsification]. AB - Clinical estimation of different modifications of phacoemulsification revealed the formation of the second tunnel in the nucleus for its division into quadrants in "four-quadrant phaco" increases the required duration of ultrasonography (US) and irrigation, which causes greater endothelial losses associated with the use of nuclear breakdown by means of a chopper tunnel. When the authors used their own methods of "extracapsular half-nuclei" fragmentation, endothelial losses are rather greater than those with the similar method "stop & "chop", which is associated with closer disposition of the US tip to the posterior corneal surface. At the same time nuclear breakdown by means a chapper in the capsular sac by the "stop & chop" method causes dilation of Zinn's ligaments, fraught by their rupture, particularly if latent derangement or defects of the zonular apparatus, and increases the risk of damage to the posterior capsule by the chopper. PMID- 12532740 TI - [Pathogenetic basis for treatment of primary keratoconus by a combined method of excimer laser surgery (combination of photorefraction and phototherapeutic keratectomy)]. AB - Bearing in mind that the pathological process in keratoconus starts with anterior corneal layers and is gradually progressing into inner corneal parts we put a question whether a pathological process can be stopped by affecting the anterior layers of the cornea. This paper presents the results of 5-year experience excimer laser surgery, a combination of photorefractive and phototherapeutic keratectomy (PRK + PTK) in the treatment of the initial keratoconus. The diagnosis of early keratoconus was based upon computed tomographic data, pachymetry in 5 points, and biomicroscopy of the cornea. The "firework" symptom a stromal ramification area that corresponds to the future apex of the keratoconus is described. PRK was performed with an ablation zone of 6 mm, and the transitional zone of 7 mm. The ablation zone in PhTk was dislocated towards the most ectasia--conical apex, topography and the apex site were predetermined of computed tomography. The PTK was performed with ablation zone of 8.00 mm and transitional zone of 9 mm. The described method allowed the authors to increase average uncorrected acuity of vision from 0.07 +/- 0.003 to 0.76 +/- 0.03. At the same time the acuity of vision was 1.0 in 69.2% of the eyes to enhance the mean corrected acuity of vision from 0.70 +/- 0.03 to 0.83 +/- 0.04. 2 by gradually progressing significantly to decline the degree of ametropia. The spherical component of refraction--myopia--decreased from 5.32 +/- 0.62 to 1.55 +/- 0.3 D. The cylindrical component of refraction myopic astigmatism reduced from 3.25 +/- 0.53 to 1.75 +/- 0.25 D. 3 to stop the progression of keratoconus in 91.3% of cases at a follow-up of 40.8 +/- 1.5 months (3 years 4 months) and at the maximal follow-up of 5.5 years. Thus, PRK + PTK with pathogenetically based, effective method in the treatment of the patients with initial keratoconus. PMID- 12532741 TI - [Prospects for treating intraocular traumatic hemorrhage using a new fibrinolytic hemase]. AB - Therapy of intraocular traumatic hemorrhage with the new drug hemaz is an important problem of ophthalmic traumatology. Seventy patients (76 eyes) with intraocular hemorrhages of traumatic origin were treated. Forty four patients (44 eyes) presented with hyphemas and 32 (32 eyes) with hemophthalmia. The results of hemaz therapy were compared with those of traditional treatment. The results confirm the high efficacy of hemaz which was well tolerated with sub-conjunctival and parabulbar injection. Indications for the use of this drug in clinical practice were formulated: it is recommended for the treatment of posttraumatic and postoperative hyphemas of any volume, including pupil hyphema, and for partial posttraumatic hemophthalmia. PMID- 12532742 TI - [Plasma-modified silicone graft for scleroplasty in progressive myopia]. AB - Plasma-chemical modification of synthetic transplant material (proliferous silicone) intended for scleroplasty in progressive myopia was theoretically substantiated, developed in the experiment and tested in practice. This treatment was shown to enhance the biocompatibility of a graft which tightly joins the sclera through the connective tissue intergrown from the surrounding capsule into the graft. The efficacy of the proposed plasma-modified grafts in stabilizing progressive myopia in 18 patients operated on (33 eyes) aged 6 to 49 years (spherical equivalent myopia degree 3.0-28.0 D) during 3- year followup was 97%. There was an elevation of uncorrected and corrected visual acuity, are active course of the postoperative period. PMID- 12532743 TI - [Transpupillary thermotherapy of classic choroidal neovascular membranes]. AB - Whether the efficacy of treatment of classic choroidal neovascular membranes (CNM) can be increased by transpupillary thermotherapy (TTT) was studied. TTT was performed by using diode laser at a wave length of 0.81 microgram. 21 patients (24 eyes) with CNM in myopia, retinal angioid streaks, age-related macular degeneration and idiopathy were followed up. The results were as follows: within 7.5 months after treatment visual acuity increased or did not change in 18 patients (21 eyes--87.5%), regression of CNM was observed in 17 cases (18 eyes- 75%). TTT in most of cases led to the regression of classic CNM without decline of visual functions. The efficacy of TTT and remote results of treatment require further investigation. PMID- 12532744 TI - [Comparative evaluation of the effectiveness of corticosteroids and cytostatics in treating retinal angiitis in Behcet's disease]. AB - Posterior uveitis and retinal vasculitis (RV) are the main causes of severe loss of vision or blindness in patients with Behcet's disease (BD). Treatment of RV in BD is a difficult problem, because the agents are effective in posterior uveitis and ineffective in RV. We observed 26 RV BD patients (20 men and 6 women aged 18 46 years, mean age 30.8 +/- 6.5 years), who were treated with corticosteroids and cytostatic drugs. Corticosteroids were used by pulse therapy, orally, and locally (periocular injections) during the active period. The results of pulse-therapy or oral corticosteroids were better than of periocular injections (92.8, 90.9, and 45.5%, respectively, p < 0.009). The velocity of reduction of inflammation and the degree of visual acuity improvement were better in the patients who received steroid pulse-therapy (12.7 +/- 2.4 and 0.19 +/- 0.21 days, respectively) in comparison with those who received oral steroids (17.7 +/- 1.8 days, p < 0.000, and 0.08 +/- 0.07, p = 0.0121). A combination of cyclophosphamide (200 mg/week intravenously) with prednisolone (10-15 mg/day) and cyclosporin (3.5 mg/kg/day) with prednisolone (10-15 mg/day) were effective for long control of RV BD patients. PMID- 12532745 TI - [Effectiveness of treating peripapillary geographic choriopathy]. AB - The results of treatment of 77 patients (127 eyes) with peripapillary geographic choriopathy (PGC) were generalized. The drug therapy gave positive results in 45.4% of cases, relative stabilization was noted in 54.6% of cases. In patients with stages I-II PGC in comparison with stage III the efficacy of treatment was significantly higher. Improvement in the exudative phase of PGC was achieved 2.6 times frequently than in the cicatricial phase. In the patients who received 2-3 courses of therapy annually the likelihood of persistence of the stabilization of the disease is 91.2%, while without treatment it descends to 29.7%. In argon laser coagulation (ALC) the positive result was obtained in 17 out of 26 eyes and retained more than 2 years in 15 out of 23 eyes. After choreoid revascularization (CR) in patients with stages II-III PGC (with visual that in those untreated functions disorders) improvement continued for 17(10 months in all 6 eyes. Operative treatment of retinal detachment in patients with PGC is effective. The findings suggest the higher likelihood of persistence of stabilization in cases of treatment in patients treatment. PMID- 12532746 TI - [Pathomorphological features of proliferative vitreoretinopathy in intravitreal administration of blood mononuclear cells (experimental study)]. AB - The role of blood mononuclears in the pathogenesis of proliferative vitreoretinopathy (PVR) was studied on 25 Wistar rats (200-250 g). Intravitreal injections of 0.05 ml normal saline containing mononuclears (3000/mm@3) into one eye and 0.05 ml normal saline into the other were made through the flat part of the ciliary body under ether narcosis. Mononuclears were isolated using the ficoll-hypaque gradient. Indirect ophthalmoscopy was carried out on days 1, 3, 5, 7, 14, and 21 after injection. After each ophthalmoscopy (except day 1) 5 animals were sacrificed and both eyes were removed. The material was fixed for optic and electron microscopy. Ophthalmoscopic and histological studies showed the development of preretinal adhesions, the main sign of PVR. Bearing in mind the unidirectional flow of liquid in ocular tissues, created by the pressure gradient, the authors suggest that blood mononuclears are involved in the development of vitreoretinal proliferation. PMID- 12532747 TI - [Prospects for radiotherapy of optic nerve meningioma]. AB - 32 patients with meningioma of the optic nerve, undergone gamma-teletherapy were followed up. The mean total focal dose was 35 Gy. 24 patients were follow-up for 1-8 years. Gamma-, therapy was shown to be the method of choice for treatment of meningiomas in the retrobulbar part of the optic nerve. It permits one to stabilize growth of the tumor, to preserve and, in number of casetelet to elevate the acuity of vision. The decline of acuity of vision was noted only in 3 cases within 6-12 months after the completion of radiotherapy. Even low initial acuity of vision and signs of atrophy of the optic nerve cannot be a ground for pessimistic prediction in relation to the results of treatment. PMID- 12532748 TI - [Use of xalatan in unstable glaucoma in myopic patients]. AB - 41 patients (79 eyes), their age averaged 66.9 +/- 4.5 years were divided into two groups. Group 1 consisted of 20 patients (39 eyes) with primary open-angle glaucoma (POAG), with refraction emmetropy (1.0 D, group 2 included 11 cases (22 eyes) in combination with myopia, group 3 included 10 patients (19 eyes) with normotensive glaucoma (NTG) and myopia. In the patients with unstabilized POAG in combination with myopia, NTG and myopia monotherapy with Xalatan (latanoprost) or combined therapy with xalatan with beta-blockers, and to a lesser extent, with Pilocarpine, was shown to stabilize the process in 35-53% of cases. PMID- 12532749 TI - [Etiology, pathogenesis and prospects for treating autoimmune eye diseases]. PMID- 12532751 TI - International journal of indoor environment and health. PMID- 12532750 TI - [Distribution and reasons for circulatory disorders in the vessels of the optic nerve]. PMID- 12532752 TI - Particle deposition indoors: a review. AB - Particle deposition indoors has received increasing attention recently because of increasing concern about the effects of particle exposure on human health. Deposition is a positive phenomenon from the perspective of human health, as deposited particles cannot be inhaled unless resuspended. There are studies showing strong correlation between the prevalence of biological-origin airborne particles and the prevalence of some specific sick building syndrome (Menzies et al., 1998; Teeuw et al., 1994). In the literature, the amount of research effort put on particle deposition indoors which is directly related to human health and microcontamination control is far less than the amount of research on deposition in small diameter tube/channel. This is a summary of the full-length version which will appear on the home page. The full-length paper aims to provide an up to-date revision for both experiment and modeling on particle deposition indoors. This paper summarizes the experimental studies for particle deposition indoors for non-industrial environments. In section 2, 'Background', an overview of the different mechanisms of particle deposition, focusing on indoor environments is addressed. In section 3, 'Experimental Study Review', a survey of aerosol deposition experiments in small experimental chambers and real houses (or large scale chambers) is presented. Detailed experimental measurements are addressed for the real house studies. The experimental techniques of particle generation, particle labeling and detection methods are discussed. Although the present paper focuses on experimental studies, it would be more complete to include some discussions on the modeling of the term particle eddy diffusivity and it is reviewed in section 4, 'Modeling Review'. PMID- 12532753 TI - Diffusion studies of VOC mixtures in a building material. AB - Volatile organic compound (VOC) adsorption by indoor materials will reduce peak concentrations, and subsequent desorption will prolong the presence of a compound indoors. Diffusion within materials plays an important role in this context. The purpose of this paper is to present two experimental set-ups to study VOC diffusion, and to show corresponding experimental results. The Field and Laboratory Emission Cell (FLEC) is in both cases an integral part of the set-up. The mass flow of VOC mixtures across gypsum board is studied. Diffusion of less polar VOCs in gypsum board can be fast (diffusion coefficients of 0.001-0.003 m2/h). Diffusion coefficients of chlorinated compounds are correlated with boiling points and molecular areas. Different compounds show similar diffusion properties. PMID- 12532754 TI - Performance and costs of particle air filtration technologies. AB - This paper predicts the reductions in the indoor mass concentrations of particles attainable from use of filters in building supply airstreams and also from use of stand-alone fan-filter units. Filters with a wide efficiency range are considered. Predicted concentration reductions are provided for indoor-generated particles containing dust-mite and cat allergen, for environmental tobacco smoke (ETS) particles, and for outdoor air fine-mode particles. Additionally, this paper uses a simple model and available data to estimate the energy and total costs of the filtration options. Predicted reductions in cat and dust-mite allergen concentrations range from 20 to 80%. To obtain substantial, e.g. 50%, reductions in indoor concentrations of these allergens, the rate of airflow through the filter must be at least a few indoor volumes per hour. Increasing filter efficiencies above approximately ASHRAE Dust Spot 65% does not significantly reduce predicted indoor concentrations of these allergens. For ETS particles and outdoor fine-mode particles, calculations indicate that relatively large, e.g. 80%, decreases in indoor concentrations are attainable with practical filter efficiencies and flow rates. Increasing the filter efficiency above ASHRAE 85% results in only modest predicted incremental decreases in indoor concentration. Energy costs and total costs can be similar for filtration using filters with a wide range of efficiency ratings. Total estimated filtration costs of approximately $0.70 to $1.80 per person per month are insignificant relative to salaries, rent, or health insurance costs. PMID- 12532755 TI - Sources of formaldehyde, other aldehydes and terpenes in a new manufactured house. AB - Formaldehyde, less volatile aldehydes, and terpene hydrocarbons are generally the predominant air contaminants in new manufactured and site-built houses. This study was conducted to identify the major sources of these compounds in a typically constructed, new manufactured house and to evaluate several source reduction practices. Specimens of materials used within the house were collected. These were individually pre-conditioned for 19 +/- 4 days, and tested for emissions of formaldehyde and other target compounds using small-scale chambers. Several cabinetry materials, passage doors, and the plywood subfloor were the predominant sources of formaldehyde and other aldehydes. The plywood subfloor was the predominant terpene source. Whole-house emission rates for combined materials were predicted based on the emission factors and the corresponding material quantities. These predicted rates were compared with whole-house emission rates derived from measurements made at the house 3 months after its installation. For 10 of 14 target compounds including formaldehyde, the predicted and derived rates were within a factor of two. Four emission barriers applied to plywood were shown to reduce emission factors for formaldehyde, hexanal, and other aldehydes. PMID- 12532756 TI - The effect of structures on indoor humidity--possibility to improve comfort and perceived air quality. AB - The research presented in this paper shows that moisture transfer between indoor air and hygroscopic building structures can generally improve indoor humidity conditions. This is important because the literature shows that indoor humidity has a significant effect on occupant comfort, perceived air quality (PAQ), occupant health, building durability, material emissions, and energy consumption. Therefore, it appears possible to improve the quality of life of occupants when appropriately applying hygroscopic wood-based materials. The paper concentrates on the numerical investigation of a bedroom in a wooden building located in four European countries (Finland, Belgium, Germany, and Italy). The results show that moisture transfer between indoor air and the hygroscopic structure significantly reduces the peak indoor humidity. Based on correlations from the literature, which quantify the effect of temperature and humidity on comfort and PAQ for sedentary adults, hygroscopic structures can improve indoor comfort and air quality. In all the investigated climates, it is possible to improve the indoor conditions such that, as many as 10 more people of 100 are satisfied with the thermal comfort conditions (warm respiratory comfort) at the end of occupation. Similarly, the percent dissatisfied with PAQ can be 25% lower in the morning when permeable and hygroscopic structures are applied. PMID- 12532757 TI - Reduced heat stress in offices in the tropics using solar powered drying of the supply air. AB - Many solutions to indoor climate problems known from developed countries may have prohibitive installation and running costs in developing countries. The purpose was to develop a low-cost solution to heat stress in a hot and humid environment based on solar powered drying of supply air. Dry supply air may facilitate personal cooling by increased evaporation of sweat. Heat acclimatized people with efficient sweating may in particular benefit from this cooling. A prototype solar powered supply system for dried-only air was made. Air from the system was mixed with room air, heated to six different combinations of temperature and humidity and led to Personal Units for Ventilation and Cooling (PUVAC) in six cubicles simulating office workplaces. A total of 123 heat acclimatized subjects were exposed 45 min in each of the cubicles. A model for the combined effect of operative temperature of room, moisture content of room air, temperature of supply air and moisture content of supply air was developed based on the experiments. Reduction of moisture content in the supply air by 1.6 g/kg had the same effect as lowering the operative temperature by 1 degree C. The solar powered system for supplying dry air is a low-cost alternative to traditional air conditioning in hot and humid regions. PMID- 12532758 TI - Cabin air quality: indoor pollutants and climate during intercontinental flights with and without tobacco smoking. AB - The aim was to determine cabin air quality and in-flight exposure for cabin attendants of specific pollutants during intercontinental flights. Measurements of air humidity, temperature, carbon dioxide (CO2), respirable particles, ozone (O3), nitrogen dioxide (NO2) and formaldehyde were performed during 26 intercontinental flights with Boeing 767-300 with and without tobacco smoking onboard. The mean temperature in cabin was 22.2 degrees C (range 17.4-26.8 degrees C), and mean relative air humidity was 6% (range 1-27%). The CO2 concentration during cruises was below the recommended limit of 1000 ppm during 96% of measured time. Mean indoor concentration of NO2 and O3, were 14.1 and 19.2 micrograms/m3, with maximum values of 37 and 66 micrograms/m3, respectively. The concentration of formaldehyde was below the detection limit (< 5 micrograms/m3), in most samples (77%), and the maximum value was 15 micrograms/m3. The mean concentration of respirable particles in the rear part of the aircraft (AFT galley area) was much higher (49 micrograms/m3) during smoking as compared with non-smoking conditions (3 micrograms/m3) (P < 0.001), with maximum values of 253 and 7 micrograms/m3. In conclusion, air humidity is very low on intercontinental flights, and the large variation of temperature shows a need for better temperature control. Tobacco smoking onboard leads to a significant pollution of respirable particles, particularly in the rear part of the cabin. The result supports the view that despite the high air exchange rate and efficient air filtration, smoking in commercial aircraft leads to a significant pollution and should be prohibited. PMID- 12532759 TI - Building-related symptoms and molds: a two-step intervention study. AB - AIMS: To evaluate the relation between mold growth and symptoms in an intervention study design. METHODS: The building was examined by a walk-through and microbiological testing from surfaces and ventilation canals before and after each of two steps of the renovation. The examination program for the 25 employees comprised questionnaire, clinical examination, 2-week peak-flow monitoring, and blood samples, and in six persons also a bronchial challenge. RESULTS: Initially the building had severe moisture problems with growth of Trichoderma and Phoma as dominating microfungi. The total number of symptoms from a nine-item building related symptom score was 66%, peak-flow variability was 20%. After the first renovation, no visible mold growth was seen, but samples showed that the building was still contaminated. Symptoms decreased to 33%. After further cleaning the mold levels decreased on surfaces, and the number of symptoms decreased to 4%. Mean peak-flow variability fell to 15%. DISCUSSION: In a poorly maintained building with moisture problems and mold growth, the staff had a number of irritative and general symptoms. The first renovation eradicated most visible signs of molds and gave a decreased number of symptoms. The second renovation sufficiently cleaned the building, and the rate of symptoms and peak-flow variability fell to normal levels. A thorough cleaning after renovation seems necessary for the eradication of symptoms. PMID- 12532760 TI - [Revision of the biological significance of the contact system]. AB - Current concept of blood coagulation is divided into two stages: an "initiation" stage which is handled by tissue factor pathway, and an "augmentation" stage handled by intrinsic pathway beginning in factor XI. Recent studies have demonstrated that the contact system is a modulator for vascular biology with vascular tone regulation, anticoagulant, profibrinolytic, antiadhesive and proinflammatory functions. Changes of contact system are associated with sepsis, thrombosis, etc. PMID- 12532761 TI - [The signal transduction and regulation mechanism of interleukin-1 on embryo implantation]. AB - This article elaborates the signal transduction pathway of interleukin-1 (IL-1) and its regulatory mechanism on embryo implantation. IL-1 is an important factor, but not the sole determinant in the regulation of embryo implantation. The cytokines are also crucial on embryo implantation. PMID- 12532762 TI - [Signal transduction pathways in cardiac myocyte hypertrophy]. AB - Cardiac hypertrophy is the consequence of hypertrophic stimulus-induced changes in gene expression, which is linked by intracellular signal transduction. It is likely, however, that there are "molecular phenotypic" differences underlying cardiac hypertrophy triggered by different stimuli, which is caused by the different signal pathways that they initiated. Studying on the signal pathways in cardiac myocyte hypertrophy contributed to elucidate the cellular and molecular mechanisms of cardiac hypertrophy and might find some new strategies for the prevention of cardiac hypertrophy. PMID- 12532763 TI - [Research progress in natural killer cell receptors]. AB - Natural killer cell expresses two kinds of recognition receptors with contrary functions: killer cell activatory receptors (KAR) and killer cell inhibitory receptors (KIR). KIR can interact with MHC I type molecular-self nonself peptide complex expressed on self cell surface, and produce inhibitory signals which can prevent the activation of KAR, thereby preventing the cytotoxicity of NK cells. NK cell will trigger effector functions(cytolysis) by KAR, if target cell loses appropriate ligands recognized by KIR. In this article we will review the research progress in the structure of these receptors, their recognition and signal transduction mechanisms. PMID- 12532764 TI - [Regulation of SWS by hormones and cytokines]. AB - SWS is the most important component of sleep. (1) VLPO-TMN seems to generate sleep and wakefulness. The rostral basal forebrain, which was defined as PGD2 SPZ, may be involved in regulation of sleep. (2) PGD2 promotes sleep, especially SWS, while PGE2 prolongs wakefulness and depresses both SWS and REMS. (3) During SWS the activation of hypothalamus-pituitary-adrenocortic axis is inhibited, while the release of growth hormone is accelerated. The soporific effects of melatonin may be attributed to its hypothermic effects. (4) Interleukin-1 prolongs sleep, especially SWS, which seems to be mediated by PGD2. Tumor necrosis factor (TFN) may promote SWS through 5-HT and its receptor. Therefore, the development of new hypnotics, which selectively prolong SWS, might follow the following ways: PGD2 and chemicals which act like PGD2; immuno-regulators; substances with effects on 5-HT receptors; hormone, such as melatonin and growth hormone, which play roles in the physiological regulation on sleep-wakefulness. PMID- 12532765 TI - [A brief introduction to the methods for novel gene cloning]. AB - There are a lot of methods for novel gene cloning, but how to clone candidate gene(s) quickly and correctly? This is a brief introduction to methods of novel gene cloning, these methods includes: differential display reverse transcriptase polymerase chain reaction(DD RT-PCR), suppression subtractive hybridization(SSH), RNA arbitrarily primed PCR(RAP-PCR), representational difference analysis(RDA), yeast two-hybrid system, cDNA capturation, et al. We not only introduced these methods, but also discussed the advantages and disadvantages of them. However, no single method is omnipotent, one should pick up the method most suitable for a special purpose. PMID- 12532766 TI - [Experimental investigation on the etiology and pathogenesis of pituitary prolactin-secreting tumor]. AB - Male SD rats transplanted with an extra pituitary in the renal capsule and treated chronically with 17-beta-estradiol (E2) were used in the studies on the pathogenesis and the underlying mechanisms of pituitary prolactin-secreting tumor (prolactinoma). The results indicated that after long treatment with E2, prolactin-secreting tumor was generated in both eutopic and ectopic pituitary, accompanied by hyperprolactinemia and overexpression of PRL gene. No apparent difference was observed in histology and ultrastructure between them. Further investigations showed that some growth factors might be involved in the tumorigenesis of prolactinoma in vivo and in vitro. A point mutation was found in the proximal promoter of PRL gene only in eutopic prolactinoma by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP), suggesting that the base change in the promoter may be related to the overexpression of PRL gene. This hypothesis has been confirmed by the increased activity of luciferase reporter fused to the mutant promoter in vitro. The coincidence of overexpression of PRL, TGF alpha and TGF beta 1 gene, and the point mutation detected in eutopic pituitary prolactinoma suggested that neuro-endocrine-immune interactions in vivo might be associated with pituitary prolactinoma formation. The mechanisms mediating tumorigenesis of eutopic and ectopic prolactinoma, respectively, may be different. PMID- 12532767 TI - [Orexin--a view discovery in obese research]. PMID- 12532768 TI - [Effect of DNA methylation on malignancy]. PMID- 12532769 TI - [The role of alpha-melanocyte-stimulating hormone in neuroimmunomodulation]. PMID- 12532770 TI - [Role of beta gamma subunits in heterotrimeric G protein signaling pathway]. PMID- 12532771 TI - [Interaction between nitric oxide and cytokines]. PMID- 12532772 TI - [Estrogen and Alzheimer's disease]. PMID- 12532773 TI - [Vascular endothelial growth factor and nitric oxide]. PMID- 12532774 TI - [Mechanisms of the suppression of hypoxic pulmonary vasoconstriction by chronic hypoxia]. PMID- 12532775 TI - [Prostaglandin and their receptors in mammalian reproduction]. PMID- 12532776 TI - [Review on the study of human and mammalian stanniocalcin]. PMID- 12532777 TI - [Phycobiliprotein and fluorescence immunological assay]. PMID- 12532778 TI - [Arterial blood pressure and its regulation]. PMID- 12532779 TI - [Voltage-gated Na+ channel in nociceptive afferents of DRG neurons]. AB - This communication briefly summarizes the molecular characteristics, expression, electrophysiological and pharmacological properties of voltage-gated sodium channels in dorsal root ganglion (DRG) neurons, and the effects of voltage-gated sodium channels on regulation of nociceptive afferents. PMID- 12532780 TI - [Neural and hemodynamic mechanisms of neurogenic pulmonary edema]. AB - Acute pulmonary edema has been reported in man and animals with intracranial disorders, head trauma or cerebral compression. In anesthetized rats, cerebral compression produced acute, fulminating and fatal lung injury. Intense activation of the sympathetic mechanism in the brain stem induced systemic hypertension and a series of hemodynamic changes. Pulmonary volume loading was the result of drastic decrease in aortic flow accompanying a decline in pulmonary arterial flow. The acute increase in pulmonary blood volume caused severe rises in pulmonary arterial and venous pressures leading to disruption of lung vessels. PMID- 12532781 TI - [Blockade of opioid tolerance and dependence by NMDA receptor antagonist]. AB - The present review summarizes the effects of drugs that block N-methyl D aspartate receptor complex (NMDA-RC) on the development of opiate tolerance and abstinence. Using behavioral pharmacological approaches, pre-clinical studies demonstrate that noncompetitive and competitive NMDA receptor antagonists and glycine binding site antagonists can inhibit opioid tolerance and some symptoms of opioid withdrawal. The biochemical mechanisms underlying the inhibition of NMDA receptor antagonists on opioid tolerance and dependence are discussed. There is some extent of neurotoxicity associated with the therapeutic use of NMDA receptor antagonists. PMID- 12532782 TI - [The effect of glucocorticoid on neural system development]. AB - During the early developmental period of central nerves system, glucocorticoid can influence the activity of hypothalamic-pituitary-adrenocortical axis via glucocorticoid receptors, and modulate the process of neural survival, outgrowth and programming death process; In adult, glucocorticoid can regulate the expression of the factors which correlate with the plasticity of neurons; In aged individual, glucocorticoid can be harmful to nerves under most conditions. Therefore, glucocorticoid plays an important role in the development of neural system. PMID- 12532783 TI - [Advances in the study of mechanisms of the tumor angiogenesis inhibitors]. AB - Tumor angiogenesis inhibitor is the drugs which can destroy or inhibit neovascularization, and block the growth and metastasis of cancer. For clinical applications it may be useful to divide the antiangiogenic drugs into two categories: class 1(specific inhibitors) and class 2 (non-specific inhibitors). This review intends to provide the recent progress of the mechanisms of antiangiogenic drugs (1) Regulation of angiogenesis growth factors; (2) Inhibition of basement membrane degradation; (3) Mediating signal transduction pathway; (4) Effect of cell cycle; (5) Regulation of tumor-related gene. PMID- 12532784 TI - [Research on the mechanism of neuronal apoptosis in Alzheimer's disease and the effects of tetrohydroberberine on the apoptosis]. AB - Apoptosis in Alzheimer's disease(AD) was studied by the techniques of immunohistochemistry, neuronal culture and patch clamp. The hippocampal neuronal apoptosis induced by beta-amyloid peptide (beta-AP) and its ionic mechanism were also observed. The results showed that neuronal apoptosis did exist in AD brains and beta-AP might induce the apoptosis by blocking the voltage-dependent K+ and Na+ channels, which were enhanced by nitric oxide(NO). In addition, tetrohydroberbine could protect against the apoptosis induced by beta-AP by reducing hippocampal neuronal intracellar free Ca2+. PMID- 12532785 TI - [In vitro and in vivo studies on the biologic effects and molecular mechanism of recombinant RA538 and antisense C-myc adenovirus on human gastric, esophageal and cancer cell lines with high-expression of Bcl-2 gene]. AB - In this study, the biological effects and molecular mechanism of recombinant RA538 and antisense c-myc adenovirus on human gastric, esophageal, 2BS and high expression bcl-2 gene cancer cell lines were studied in vitro and in vivo. The results were as follows: Ad-RA538 and Ad-ASc-myc could strongly inhibit cell growth and induce apoptosis of SGC7901 cells in vitro and in vivo, and could down regulate expression of c-myc, bcl-2 and cyclinD1 gene, up-regulate expression of bax gene. Ad-RA538 or Ad-AS c-myc could not inhibit cell growth and induce apoptosis changes of EC109, EC8712, 2BS and high-expression bcl-2 gene cancer cell lines, and could not down-regulate expression of c-myc and bcl-2 gene. The results indicated that: Ad-RA538 or Ad-AS c-myc can inhibit growth and induce apoptosis of gastric cancer cell in vitro and in vivo. They relate to c-myc, bcl 2, cyclinD1 and bax gene closely and play a key role on biologic effects in gastric cancer cells. Ad-RA538 and Ad-AS c-myc could not produce relevant changes on esophageal cancer, 2BS and high-expression bcl-2 gene cell lines. PMID- 12532786 TI - [Bovine spongiform encephalopathy: an autoimmune hypothesis]. PMID- 12532787 TI - [Adrenomedullin receptors and its signal transduction]. PMID- 12532788 TI - [The rostral medullary botzinger complex and respiratory rhythm formation]. PMID- 12532789 TI - [Connexin gene mutation and disease]. PMID- 12532790 TI - [The structural features and inhibitors of tumor necrosis factor-alpha converting enzyme]. PMID- 12532791 TI - [Progress in the study on brain mechanisms of priming effects]. PMID- 12532792 TI - [Relation between rheumatoid arthritis and interleukin-1]. PMID- 12532793 TI - [The cellular signal transduction of TGF-beta family]. PMID- 12532794 TI - [Plasticity change and functional reorganization in the brain]. PMID- 12532795 TI - [Developmental plasticity in neural circuits for song learning of birds]. PMID- 12532796 TI - [Advances in the research of osteopontin]. PMID- 12532797 TI - [Representational difference analysis in cloning of aberrant genes]. PMID- 12532798 TI - [O6-methylguanine-DNA-methyltransferase and prediction of cancer chemotherapy]. PMID- 12532799 TI - [Internet and development of nutrition]. PMID- 12532800 TI - [Coding theories and analyzing methods of information contained in neuronal spike trains]. AB - Information transmission in the central nervous system relies on neurons working in relays. At chemical synapses information transmission is by means of neurotransmitters, whereas long-distance transmission is carried out by action potentials conducting along nerve fibers. Hence how to pick up and analyze information contained in the spike trains are important topics of several branches of sciences such as neuroscience, bioinformatics, etc. So far there is lack of ideal ways and means. This review briefly introduces the advantages and limitations of the principal coding theories and analyzing methods of information contained in spike trains, and the progress in this field. PMID- 12532802 TI - [On the mechanisms of cardiac hypertrophy]. AB - Here we reviewed the progress of researches on the mechanism of cardiac hypertrophy and discussed its putative mechanisms. With regard to the heredity and variability of familial hypertrophic cardiomyopathy and mitochondrial cardiomyopathy and other changes, we emphasize that the imbalance between the energy supply and demand of the overloaded pressure determines the occurrence of cardiac hypertrophy. Also we refer to the involving roles of the mechanical stress and some paracrine/autocrine factors in cardiac hypertrophy. Finally we hypothesize that there exists an unknown small molecule being involved in regulating the gene expression and phenotype in cardiac hypertrophy. PMID- 12532801 TI - [Subtypes and functions of voltage-gated Na+ channel]. AB - Since 1990s, the research on voltage-gated sodium channels have got rapid progress by the use of molecular biological and electrophysiological recording techniques. This communication presents a brief review on mammalian sodium channel involving classification, distribution, and mechanisms of expression regulation of various subtypes in the central and peripheral nervous system and other nonexcitable tissues. PMID- 12532803 TI - [Phospholipase D and inflammation]. AB - Phospholipase D (PLD) is widely distributed in mammalian cells, where it is regulated by a variety of extracellular signals. Its major substrate is phosphatidylcholine (PC), which is hydrolyzed to phosphatidic acid (PA) and choline. The PLD-catalysed PC hydrolysis is believed to be an important mechanism of signal transduction in cells. More and more lines of evidence indicate that PLD is tightly involved in inflammation. This review introduces the recent advances of PLD in respiratory burst, degranulation and arachidonic acid release. PMID- 12532804 TI - [Calcium and chronic diseases]. AB - Calcium plays a role in physiological functions in our body. Many diseases are associated with physiological dysfunction caused by abnormal calcium homeostasis. In this review, we discussed the relationships between calcium and some common chronic diseases including cardiovascular diseases. Here we put emphasis on hypertension and atherosclerosis, tumor, osteoporosis and osseous hyperostosis. The most appropriate daily calcium intake and adverse effects of high-calcium diets are also mentioned in this article. PMID- 12532805 TI - [Protection from oxidant injury on airway epithelial cells and the local microenvironment modulation]. AB - An cellular injury model of primary cultured rabbit bronchial epithelial cells (BEC) exposed to ozone was established in this study and cytoprotective effects of factors in local microenvironment of airway such as vasoactive intestinal peptide (VIP), epidermal growth factor (EGF) and heat stress were observed. These factors could lighten damage in cell and elevate the level of glutashione(GSH), which depended upon phosphorylation modulation by protein kinases and gene transcription. A low level of bcl-2 gene expression could be detected in BEC at basic state, and either VIP or EGF stimulated the transcription of bcl-2, which improved the capability of BEC to resist oxidant injury. Otherwise, EGF and heat stress increased VIP autocrine from BEC and upregulated the expression of VIP receptor on BEC, so that the protective effect of VIP can be amplified in local sites. This study confirmed that there is an anti-injury protection on airway epithelium and the protection can be adaptively modulated by the regulatory peptides in local microenvironment and exogenous stimulus. PMID- 12532806 TI - [Studies on the effects of astrocyte during glial scar formation by recombinant antisense GFAP retrovirus]. AB - The present study was to investigate the changes of macroglia and the effect of gonadal hormone on reactive gliosis and of antisense GFAP retrovirus on astrocyte, gliosis and glial scar formation after brain stab injury (BSI). The results indicate that astrocytes are the main cells of glial scar. GFAP plays an important role in the maintenance of structure and function of astrocytes. The oligodendrocyte is not an active cell during glial scar formation. The gonadal hormone can modify the reactivity of astrocyte after BSI, but has no significant effect on differentiation and proliferation of astrocyte. The recombined GFAP retrovirus can effectively inhibit the growth of astrocyte, and GFAP expression of injured astrocyte in vitro and glial scar forming in vivo. PMID- 12532807 TI - [Signaling pathways of lysophosphatidic acid]. PMID- 12532808 TI - [Progress in calcium signaling of cardiac muscle cells]. PMID- 12532810 TI - [Signal transduction system of cellular nuclear reaction caused by extracellular signals]. PMID- 12532809 TI - [Cytochrome C and cells apoptosis]. PMID- 12532811 TI - [Nucleic acid receptor and its effect]. PMID- 12532812 TI - [Sodium-calcium exchange currents in cardiac myocytes]. PMID- 12532813 TI - [Capillary gel electrophoresis in clinical molecular biology]. PMID- 12532814 TI - [Effect of alpha-interferon in the central nervous system]. PMID- 12532815 TI - [The way of disparity encoding in visual system in stereopsis--position shift? Phase shift?]. PMID- 12532816 TI - [Parkinson's disease and oxidative stress]. PMID- 12532817 TI - [Apoptosis in the nervous system]. PMID- 12532818 TI - [Experimental studies and clinical application of a low molecular weight tumor suppressor on selective eliminating of leukemia cells]. PMID- 12532819 TI - [Nitric oxide and its enlightenment]. PMID- 12532820 TI - [Microsatellite instability: a potential tool for the study of life sciences]. AB - Microsatellites are simply repeated nucleotide sequences scattered throughout the human genome. They are highly polymorphic among human population and inherited in a stable manner. The microsatellite instability (MI) is highly polymorphic, which is associated with the defects in DNA mismatch repair genes. MI has been widely used by scientists to study the tumorigenesis. On the basis of their findings, a "mutator that mutates the other mutator" model for tumorigenesis has been proposed. MI is also a potential tool for the study of genetics, aging and other life sciences. PMID- 12532821 TI - [Channelopathy of voltage-gated sodium channel]. AB - This review will give a brief introduction on "Channelopathy" of voltage-gated sodium channel involved with inherited diseases including skeletal muscle diseases (such as HyperPP and PC) and cardiac diseases (such as LQT3 and IVF), voltage-gated sodium channel mutations and the electrophysiological characteristics of the sodium channel mutations. PMID- 12532823 TI - [Spreading depression and cerebral ischemia]. AB - Growing evidence indicates that spreading depression (SD) may not only relate to some diseases, including cerebral ischemia, migraine, epilepsy, traumatic brain injury and so on, but also affect the physiological courses such as sleep patterns and general arousal. In this paper we review the relationship between SD and cerebral ischemia. The following questions are discussed: (1) The discovery of SD and its brief research history; (2) SD in the intact brain; (3) Peri infarct SD in focal cerebral infarction. It is suggested that peri-infarct SD contributes to the growth of focal ischemic lesions, and the suppression of SD may become a novel, even a key therapeutic approach for minimizing infarct volume and protecting ischemic neurons after focal cerebral ischemia. PMID- 12532822 TI - [The biological effect of tetrahydrobiopterin and its potential role in sepsis]. AB - It is well known that overproduction of nitric oxide (NO) is the final common pathway in septic shock. Tetrahydrobiopterin (BH4), a low molecular weight pterdine, is an essential cofactor required for the activity of NO synthase (NOS). Existing evidences show that lipopolysaccharide and proinflammatory cytokines can cause significant rises in bipoterin levels, which in turn augments the synthesis of NO. Also inhibition of biosynthesis of BH4 can decrease NO formation, implying that BH4 may be involved in the pathophysiological alterations of sepsis. However, the precise mechanisms of BH4 in regulating NO formation are not yet fully understood. In this review, we focus on the biological effects and regulation of BH4, as well as its potential role in sepsis. The therapeutic significance of biopterin synthesis inhibitors in septic symptoms is also discussed. PMID- 12532824 TI - [Mechanisms underlying dopaminergic neurotoxicity in ischemic cerebral damage]. AB - Dopamine(DA) is a neurotransmitter, but it also serves as a neurotoxin under certain pathological conditions. Accumulating evidences indicate that DA plays an important role in ischemic cerebral damage. The mechanism by which DA exacerbates the neuronal damage remains unclear. Many mechanisms have been proposed to explain the DA neurotoxicity. One possibility is that DA leads to the formation of free radicals which are cytotoxic. Another possibility is that direct dopamine receptor stimulation may mediate dopaminergic neurotoxicity. It is also possible that DA modulates excitotoxicity of excitatory amino acid. PMID- 12532825 TI - [Gap junctions in the nervous tissue]. AB - In recent years, prominent progresses have been made in the study of the localization and function of gap junctions in the nervous tissue. The application of molecular approaches has advanced the disclosure of the structure, classification and biophysical properties of gap junctions. Dye-coupling and Ca2+ imaging techniques have provided explicit and effective ways for the study of gap junctional functions. The regulation of gap junctions involves the expression of gap junction genes and changes in the conductivity of gap junction channels. Gap junctional intercellular communication includes the exchange of second-messengers and electrical coupling between coupled cells. GJ plays an important role in the function of neuroglial cells, neurons in some brain regions and probably in the formation of local neuronal circuits. PMID- 12532826 TI - [A study on the effect and mechanism of acupuncture suppression of neuronal apoptosis following cerebral ischemia]. AB - On the basis of affirming that acupuncture suppresses neuronal apoptosis following cerebral ischemia, this study investigates the mechanism of acupuncture which modulates the intrinsic factors suppressing and promoting apoptosis. The main results showed that: (1) the symptom of neral defect induced by cerebral ischemia was improved and the area of infarction following cerebra ischemia was reduced after acupuncture treatment; (2) by employing PI and TUNEL staining, the apoptosis neurons were observed following cerebral ischemia, and acupuncture was able to protect against neuronal apoptosis following cerebral ischemia; (3) using a kinetic cadmium-reduction method for the determination of nitric oxide (NO) and an immunohistochemistry method for observation of NOS-immunoactivity in rat brain, it was found that the content of NO in the side of infarction was higher than that of control, and the increase of NO content was positively related to iNOS immunoactivity; acupuncture could inhibit iNOS activity thereby decreasing the NO level; (4) in situ hybridization study revealed that excitotoxicity of glutamate following cerebral ischemia was mediated by NMDAR1mRNA overexpression, that was down-regulated by acupuncture; (5) immunohistochemistry study revealed that the expression of nerve growth factor receptor(trk-A) was induced by acupuncture after cerebral ischemia, which may play a role in antagonizing apoptosis. PMID- 12532827 TI - [Modulatory effects of capsaicin-sensitive nerve fiber on cardiovascular activity]. PMID- 12532828 TI - [Research progress on the effect of zinc on behavior]. PMID- 12532829 TI - [Iron, transferrin and transferrin receptor in the brain]. PMID- 12532830 TI - [Relation between the properties and its induced signal transduction pathways of muscarinic acetylcholine receptor and neoplasm]. PMID- 12532831 TI - [Research progress on the signal transduction and the molecular regulation of oocyte meiotic maturation]. PMID- 12532832 TI - [P2X purinoceptors and their regulations]. PMID- 12532833 TI - [Recent research advances in oxidized high density lipoproteins]. PMID- 12532834 TI - [The correlations between glucocorticoid receptor of lymphocyte and immune function]. PMID- 12532835 TI - [Protein kinases and Alzheimer's disease]. PMID- 12532836 TI - [Mitochondrial defects and Alzheimer's disease]. PMID- 12532837 TI - [Focal adhesion kinase]. PMID- 12532838 TI - [Progress in the study on cyclic ADP-ribose]. PMID- 12532839 TI - [Mechanism of modulation of synaptic efficacy by stress and learning]. PMID- 12532840 TI - [Nomenclature and abbreviation in nitric oxide research]. PMID- 12532841 TI - [The key enzyme of cholesterol synthesis pathway: HMG-CoA reductase and disease]. AB - In eukaryotes, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase is a key enzyme that catalyses the synthesis of a precusor of cholesterol as well as non sterol isoprenoids, mevalonate. The regulation of the enzyme activity occurs at the transcription, post-transcription, translation and protein degradation levels. Cholesterol contributes to the generation and development of atherosclerosis while non-sterol isoprenoids play a role in regulation of cell proliferation, signal transduction and generation of cancers. At present, the enzyme is the target of several drugs effective against atherosclerosis. PMID- 12532842 TI - [Response kinetics of GABA receptors and their functional significance]. AB - It is an important issue whether and how the response kinetics, including activation, desensitization and deactivation, of gamma-aminobutyric acid (GABA) receptors plays roles in rapid inhibitory postsynaptic transmission. With the development of molecular biology, and the use of patch clamp techniques and rapid solution changers, the structural basis, underlying mechanisms at single channel level and functional roles of the characteristics of GABA receptors have been fully explored. It is revealed that the response kinetics of GABA receptors plays a crucial role in modulating inhibitory postsynaptic currents and has a great importance for rapid inhibitory synaptic transmission. PMID- 12532843 TI - [Roles of alpha-2 adrenoceptor in prefrontal cortical cognitive functions]. AB - Studies in non-human primates have demonstrated that norepinephrine can improve, through actions at postsynaptical alpha-2A adrenoceptors, cognitive functions of the prefrontal cortex, such as attentional regulation, working memory and response inhibition. Several psychiatric disorders, including attention-deficit hyperactivity disorder (ADHD), show prominent syndromes seen in cognitive deficits of the prefrontal cortex. These basic studies help develop novel pharmacotherapies for psychiatric disorders such as ADHD. PMID- 12532844 TI - [Relationship between oxysterols and atherosclerosis]. AB - Oxysterols, being oxygenated derivatives of cholesterol, are as many kinds as one hundred. They are found in the foodstuffs, the blood and arterial tissues of animals with hypercholesterolemia, the human atheroma, the foam cell from atherosclerotic tissues, as well as in the oxidized low density lipoprotein. The results of experiments have demonstrated that oxysterols are cytotoxic and they are injurious to a variety of cells including vascular endothelial cells, smooth muscle cells and monocyte/macrophages, suggesting that oxysterols could play a key role in the initiation and pathogenesis of atherosclerosis. PMID- 12532845 TI - [The signal transduction of cell activation by LPS: the studies from CD14 to p38 MAPK]. AB - Recently there have been some substantial progresses in signal transduction of cell activation by lipopolysaccharide (LPS). After binding with LBP in the plasma, LPS is transported to the cell membrane surface of monocytes, then binds with membrane CD14 to induce cell activation. Mitogen-activated protein kinases (MAPKs) take part in the signal transduction of LPS stimulated cell activation to produce bioactive substances including tumor necrosis factor. p38 MAPK is an important regulator for TNF-alpha expression. It is possible to provide new theory and idea for the treatment of septic shock from the studies of signal transduction of cell activation by LPS. PMID- 12532846 TI - [Repeated 100 Hz TENS for the treatment of chronic inflammatory pain in rats: optimal parameters and possible neuro-chemical mechanisms]. AB - AIMS: To establish a reliable animal model of chronic arthritic inflammatory pain, to find the optimal parameters and appropriate intervals of repeated 100 Hz transcutaneous electrical nerve stimulation (TENS) for the treatment of the chronic arthritic pain, and to explore the possible central neurochemical mechanisms. THE RESULTS: (1) Injection of water-in-oil type complete Freund's adjuvant into the tibio-tarsal joint of the rat produced a reliable, reproducible monoarthritic model. (2) The interval between 2 sessions of stimulations and the intensity of stimulation were important factors determining the therapeutic effects of repeated 100 Hz TENS on both acute and stable period of monoarthritis. (a) For acute period of monoarthritis, the optimal frequency is twice a week, while in the stable period, once a week seems better. (b) The effects of TENS in three intensities: the weaker stimulation was found to produce better results. (3) In the period from 2 to 9 weeks, the levels of spinal release of SP-ir in the TENS group were lower than the control group. This may constitute one of the possible neuro-chemical mechanisms underlying the therapeutic effects of multiple TENS treatment. (4) In both acute and stable period of adjuvant-induced monoarthritis, intrathecal microinjection of Nor-BNI produced dose-dependent increase of arthritic flexion pain scores (i.e. hyperalgesia), suggesting that spinal dynorphin/kappa system plays a role in suppressing arthritic pain. PMID- 12532847 TI - [The immunoregulatory effects of beta-endorphin]. PMID- 12532848 TI - [Helicobacterium pylori infection and coronary heart disease]. PMID- 12532849 TI - [Structure and function of plasminogen-related growth factor family]. PMID- 12532850 TI - [Effects of adrenoceptors on growth and apoptosis of cardiovascular cells]. PMID- 12532851 TI - [A design of a novel compound S-nitrosocaptopril possessing the capacities of both angiotensin converting enzyme inhibitors and NO donors]. PMID- 12532852 TI - [The central role of apolipoprotein E in the nervous system]. PMID- 12532853 TI - [NMDA receptor and the development of central nervous system]. PMID- 12532854 TI - [Apoptosis of skeletal muscle cell]. PMID- 12532855 TI - [Molecular mechanism of vasculogenesis and angiogenesis]. PMID- 12532856 TI - [Cellular metabolism and pathological role of peroxynitrite]. PMID- 12532857 TI - [Protein kinase C and cardioprotective effects of ischemic preconditioning]. PMID- 12532859 TI - [Progress in the study of cis-9,10-octadecenoamide]. PMID- 12532858 TI - [The research progress on preprotachykinin]. PMID- 12532860 TI - [Functional magnetic resonance imaging of human brain and its uses in the study of recognition in neurology]. PMID- 12532861 TI - [Psoriasis treatment update]. PMID- 12532862 TI - [Popper, Duhem and clinical trials: are biomedical laws falsifiable?]. PMID- 12532863 TI - [Pathogenesis of scleroderma]. AB - Systemic sclerosis is a systemic inflammatory disorder, affecting the skin and other organs, especially the lungs and the digestive tract. The pathogenic mechanisms of systemic sclerosis still remain unknown, although there is evidence that many parameters may play a role in the pathogenesis of the disease, including: vascular involvement, immunity impairment, and both environmental and genetic factors. To date, many authors support the hypothesis that, in patients with genetic predisposition, environmental stimuli (occupational factors and/or virus) may lead to vascular dysfunction, particularly, endothelial damage and immune activation, resulting in tissue fibrosis at late stages of the disorder. PMID- 12532864 TI - [How to recognize scleroderma]. AB - Systemic sclerosis is a multisystem disorder characterized by the presence of connective tissue sclerosis, vascular abnormalities concerning small-sized arteries and the microcirculation, and autoimmune changes. This disease is usually preceded by Raynaud's phenomenon. Cutaneous changes are useful for diagnosis. Sclerosis is initially proeminent on hands with frequently pitted scars. Telangiectases and calcium deposits are mainly observed in a good prognosis subset of the disease (CREST) characterized by the association of limited skin involvement and anti-centromere antibodies with possible late onset of pulmonary hypertension. The bad prognosis subset is associated with diffuse cutaneous sclerosis, pulmonary fibrosis, gastrointestinal disease, myocardial involvement and anti-topo-isomerase I antibodies. Histological skin changes are similar to those observed in morphea. In morphea, there is only localized sclerosis of the skin with highly variable aesthetic disability. The condition may be subdivided clinically into the following types: circumscribed plaques, linear morphea and frontoparietal lesions. The last two forms may be associated with atrophy of the underlying anatomic structures. Many other diseases may have a scleroderma-like appearance. In these diseases, nail folds abnormalities, visceral involvement and immunological changes are usually different from those observed in systemic scleroderma. PMID- 12532865 TI - [Sclerodermic acrosyndromes]. AB - In patients with systemic sclerosis, two main subsets of vascular acrosyndromes can be found: Raynaud's phenomenon and digital ulcerations. A severe Raynaud's phenomenon is found in almost every patient with systemic sclerosis. Most often, it is the first sign announcing the emergence of the disease and the hypothesis of subclinical scleroderma should be evaluated in every patient with apparently isolated Raynaud's phenomenon. Digital ulcerations can show several clinical aspects: ischemic micro-ulcerations and pitting scars of the finger tips, nailbed and nailfold lesions and digital necrosis, but also chronic ulcerations of the dorsal skin surface of the finger joints and ulcerated dermal calcifications, the origin of which is not only ischemic. First line treatment of such acro-syndromes is based upon cold avoidance and calcium-blocking agents. In the most severe cases, hospitalization in specialized centers is required, where customized physiotherapy and perfusions of prostaglandins or dextrans will be performed. PMID- 12532866 TI - [Digestive involvement of scleroderma]. AB - Digestive involvement in systemic sclerosis is frequent and serious, because it provides morbidity and fatality. From the pathophysiologic point of view, the first step could be Raynaud-associated neural dysfunction, followed by smooth muscle atrophy then irreversible muscle fibrosis. Oesophageal disorder is common with its main consequence: the occurrence of gastroesophageal reflux disease which could run into peptic erosive oesophagitis. Oesophageal manometry is the main diagnostic tool, gastrointestinal endoscopy helps to assess oesophageal mucosal inflammation and its possible sequels. Gastric involvement is rarely recognized but it is frequent in case of systematic investigation as well as small intestinal involvement which may provide a lot of complications: malabsorption, pseudoobstruction, bacterial overgrowth. At colonic level, anorectal involvement is frequent and leads to fecal incontinence and rectal prolapse. Reynold's syndrome is a special case which associates systemic sclerosis with primary biliary cirrhosis. The symptomatic treatments must be systematic and improve the disease's overall prognosis. PMID- 12532867 TI - [Cardiac and pulmonary involvement in scleroderma]. AB - Systemic sclerosis is a connective tissue disease characterised by microvascular involvement and fibrosis affecting skin, lung, heart and intestinal tract. Sixty percent of deaths are secondary to heart or lung involvement and (or) to pulmonary hypertension. Clinical expression of these complications is present only in patients with the most severe problems. Therefore, it is appropriate to systematically and repeatedly look for these complications, even in asymptomatic patients. Heart involvement can present as cardiomyopathy, pericarditis, conduction or repolarisation abnormalities and arrhythmia, sometimes leading to sudden death. Pulmonary hypertension is due either to direct involvement of pulmonary arteries or is secondary to lung fibrosis. Doppler echocardiography and lung function tests are systematically and repeatedly proposed to screen patients for pulmonary hypertension at initial diagnosis and during follow-up. Right heart catheterisation ascertains the diagnosis of pulmonary hypertension. Chronic pulmonary interstitial fibrosis is the most frequent manifestation of lung involvement. High resolution computed tomography is the method of choice to make the diagnosis and to assess the severity of the lesions. Treatment is not completely standardised. However, a severe and rapidly progressing lung involvement is probably a good indication of immunosuppressive agents, such as cyclophosphamide. PMID- 12532868 TI - [Renal involvement in scleroderma]. AB - Scleroderma renal crisis occurs most often during the first years of the disease, in patients with systemic sclerosis and evolving cutaneous lesions. Clinically, it is responsible for severe hypertension, sometimes associated with cardiac failure or neurological symptoms. Laboratory tests disclose rapidly progressive renal failure, and often signs of thrombotic microangiopathy. If performed (which is rarely the case), renal biopsy shows scleroderma-induced chronic vascular lesions, but also vascular lesions that are secondary to malignant hypertension. The cornerstone of treatment is blood pressure control using angiotensin converting enzyme inhibitors, often in association with other antihypertensive agents. It has to be started as early as possible, in order to optimise vital and renal prognosis. PMID- 12532869 TI - [Treatment of scleroderma]. AB - No single therapeutic agent has been shown to increase survival of patients with systemic scleroderma in a prospective randomized trial. Drugs as colchicine and D penicillamine are disappointing. Conversely, disease modifying agents are emerging such as cyclophosphamide in interstitial pulmonary disease, and stem cell autograft after high dose cyclophosphamide therapy in patients who develop visceral involvement in the three first years of evolution of the disease is under study. Organ specific therapy may show significant benefit, such as angiotensin converting enzyme inhibitors in renal crisis and epoprostenol in primary pulmonary hypertension. PMID- 12532870 TI - [Living with scleroderma]. AB - To live with scleroderma is a very difficult challenge, owing to the various and sometimes painful or incapacitating consequences of the disease. The lengthy evolution, the uncertain prognosis and the functional handicap yield a complex and heavy burden on the patient and his family. Numerous means exist to counter the sequels of scleroderma, but the problem is to combine them correctly to obtain the best possible result in terms of comfort and quality of life. The various life aids, the patients association, and the non pharmaceutical treatments have an important role to play besides the intervention of the medical specialists of the organs involved by scleroderma. PMID- 12532871 TI - [Students enrolled in the first year of medical school]. PMID- 12532872 TI - [Accidents caused by anticoagulants]. PMID- 12532873 TI - [Hemorrhoidal pathology]. PMID- 12532874 TI - [Acute urinary retention]. PMID- 12532875 TI - [Dental and gingival lesions]. PMID- 12532876 TI - [Genital infections in men. Urethral discharge]. PMID- 12532877 TI - [Weight loss. Diagnostic approach]. PMID- 12532878 TI - [Central Scientific and Research Institute of Stomatology meets its 40th anniversary]. PMID- 12532879 TI - [Pathology and problems in theoretical and practical dentistry]. AB - Pathology is a science describing the common regularities of pathological processes underlying disease development. Today pathology is defined as a theoretical science, which is at the same time closely connected to clinical practice. This paper reviews the history of research activities of Laboratory of Pathology of Central Institute of Stomatology. New methods have always been used at the laboratory; today it is histochemistry, radioisotope method (specifically histoautoradiography), and electron microscopy (including cytochemistry). Many year research activities of the laboratory confirm the significance and perspectiveness of basic studies in dentistry, which are of priority importance for the development of theoretical dentistry and for solving the pressing problems in clinical practice. PMID- 12532880 TI - [Study of protective effect of perftoran on a skin flaps]. AB - Differences in the levels of tissue respiration in the rat skin flaps incubated in perfluothane (PF) and normal saline (NS) are regarded as an evidence of local PF effect on tissue substrate. The complex of metabolic changes in the skin fragments under the effect of PF in comparison with skin fragments incubated in NS indicate a protective effect of PF on tissue substrata. Decreased content of malonic dialdehyde and a slower accumulation of tissue liquid in skin flaps incubated in PF, in comparison with the fragments incubated in NS, is worthy of note. It indicates that incubation in PF (in comparison with incubation in NS) is less harmful for cell membranes and induces lesser disorders in water-salt metabolism and edema which are usually associated with tissue lesions developing under conditions of trophic deficiency. PMID- 12532881 TI - [Optimization of structural characteristics of a root canal wall during retrograde tooth filling on a basis of scanning electron microscopy data]. AB - Studies by scanning electron microscopy showed that dentin of the root stump (resection of the root apex is an obligatory condition of retrograde filling) has numerous regular dentin tubules, in contrast to the apical part of the root. Mechanical treatment of dentin during preparation of the platform for retrograde filling leads to the formation of a thick "lubricating" layer on the surface of these platform. This layer consists of corpuscles of destroyed tissue substance (smudge) and completely covers the ends of dentin tubules, thus preventing the formation of strong bonds between the filling material and dentin surface. Treatment (pickling) of the platform surface with the conditioner notably improved the adhesion of Russian filling material Dentis to dentin. The best adhesive effect of retrograde filling was attained with Kemfil, the material requiring no pickling pretreatment. PMID- 12532882 TI - [Efficiency of M-22 human fibroblast culture as a tissue engineering factor in plastic repair of mandibular bone defects]. AB - The results of inoculation of human fibroblast (strain M-22) and rabbit fibroblast cultures in standard bone defects were studied in 4 groups of rabbits 3 days to 3 months after inoculation of the cultures together with osteoplastic material (collapol KP-2). In the control groups the defects healed under a blood clot (group 1) or were filled with KP-2 (group 2). Histomorphological analysis of tissue showed that inoculation of M-22 cell culture into bone defects notably activated reparative osteogenesis in osseous wounds and stimulated healing, which was more rapid than in other groups. Inoculation of rabbit fibroblast culture in bone defects slightly inhibited the healing of bone wounds. PMID- 12532883 TI - [Caries resistance]. AB - Discussing the problems of caries resistance and caries susceptibility, we should remember that normal status of dental enamel is maintained by the dynamic equilibrium between demineralization and remineralization processes. Some aspects in the structural resistance of the enamel, determined by its maturing, are discussed. Data on the role of oral factors reflecting the status of the organism are presented. The authors do not agree that caries resistance, which is permanent throughout life, is determined by the CDL index. They give their arguments in favor of introducing the notion of "caries risk" and the criteria for evaluating this risk. PMID- 12532884 TI - [Alternative methods for prevention and treatment of dental caries using laser and magnetic laser exposure]. AB - Alternative methods for prevention and treatment of dental caries are presented, based on the use of laser and magnetic laser exposure (patent No. 2053818, in Russia) with a new generation laser device with Optodan microprocessor monitoring (patent No. 2014107, Russia). The methods are intended for wide clinical application in children and adolescents in pedodontic departments and outpatient centers and particularly for group and individual use in dental rooms at school. PMID- 12532885 TI - [Preparation of hard dental tissues by aeroabrasive method]. AB - The efficiency of aeroabrasive preparation (AAP) of teeth was evaluated by clinical methods and scanning electron microscopy. The authors conclude that the use of AAP should be confined to the treatment of surface and medium caries. Analgesia before AAP is obligatory, except cases with preparation of fissures for preventive hermetization. The use of AAP is absolutely contraindicated in cases of allergic reactions in the patient, dentist, or assistant. Methods for protection of the dentist, assistant, and patient during the use of AAP are discussed. PMID- 12532886 TI - [Long-term results of front teeth restoration using light-curing composite materials]. AB - 200 patients (500 restorations of front teeth) were examined. All the restorations were made of light-curing composite materials (microfills, hybrids, compomers, and combination of microfills and hybrids) from 5 do 9 years ago. Clinical and esthetical parameters were evaluated. We had to replace 96 restorations of 500 (19.2%), only 50 (10%) restorations were marked as "excellent". Direct veneers made of microfilled composite materials showed the best results (30%--"excellent" restorations). The restorations of devital teeth and V-class restorations of compomer material show the worse result (20% restorations replaced). PMID- 12532887 TI - [Classification of aggressive form of periodontal diseases]. AB - An attempt at objective evaluation of currently used classifications of periodontal diseases is made. The first classification of aggressive forms of periodontitis was offered in 1983. Classifications of 1982, 1992, 1999, and 2001 also reflect the clinical forms of atypical periodontitis. PMID- 12532888 TI - [Role of microflora in diseases of the buccal mucosa]. AB - The role of microflora was studied in patients with diseases of the buccal mucosa (aphthous stomatitis, lichen planus, leukoplasia, desquamative glossitis, etc.). Quantitative and qualitative composition of the microflora and its sensitivity to antibiotics, antifungal agents, and bacteriophages were studied. The following changes in the microflora were distinguished: dysbiotic shift, dysbacteriosis of the I-II, III, and IV degree. Microbiological status of patients with diseases of the buccal mucosa is essential for the course, outcome, and prognosis of the underlying diseases. PMID- 12532889 TI - [Main trends in development of craniofacial surgery for congenital defects]. PMID- 12532890 TI - [Planning of orthodontic treatment in patients with maxillary dentition defects using mathematical methods]. AB - Stress-deformed status of bone tissue in the implantation zone was studied. The relationship between the level of strain in the spongy and cortical layers of the bone and spatial location of the implant in the jaw and its geometrical parameters is demonstrated. Numerical analysis of the level of bone tissue stress was carried out on the basis of experimentally created three-dimensional solid body model of the maxilla. The process of changes in the distribution of tensions during different phases of mastication was investigated. The potentialities of numerical analysis in the planning of orthodontic treatment of patients with dentition defects making use of implants are demonstrated. PMID- 12532891 TI - [Types of impressions for permanent dentures: classification and terms]. AB - Some classifications of anatomical impressions and the relevant terms are discussed. Methods for impression making are described in brief and their main characteristics are presented, based on which these methods are classified into groups. A new classification of impressions is suggested. PMID- 12532892 TI - [Effect of selective polishing of teeth on local circulation in prosthetic treatment by implantation]. AB - An effect of occlusion disorders on a regional circulation was studied in periodontal tissues of abutment teeth, antagonist teeth, periimplant zone at different stages of preparation for a prosthesis by normalization of occlusion ratios by means of selective polishing of teeth. The terms and the periodicity of selective polishing promoting redistribution of the masticating pressure were determined. The connection of these factors was examined with periodontal vessel tonicity and periimplant zone on the basis of the rheoparodontographic findings. PMID- 12532893 TI - [Sixty years of creation of Russian acrylic materials for dentistry. History and prospects]. PMID- 12532894 TI - [Age dynamics of the prevalence and changes in the structure of maxillodental abnormalities among children of preschool age and schoolchildren]. PMID- 12532895 TI - [Central Scientific and Research Institute of Stomatology: Difficult time of choice and transfer to the marketing conditions (1986-1996)]. PMID- 12532896 TI - [Contribution of the Orthodontic Department of Central Scientific and Research Institute of Stomatology to development of this specialty]. PMID- 12532897 TI - [X-Ray Department of Central Scientific and Research Institute of Stomatology: Summing up 40-year activities]. PMID- 12532898 TI - [Postgraduate training of research, training staff, and dentists at the Central Scientific and Research Institute of Stomatology]. PMID- 12532899 TI - [Comparison of extraction methods for detection of Ehrlichia DNA in whole blood using PCR]. AB - The author compared nine methods for isolation of specific DNA of the etiological agent of human granulocytic ehrlichiosis from human blood for examination of the PCR. To full blood of healthy donors a laboratory culture of the agent was added and the effectiveness of isolation an stability of the template was tested under conditions when the blood was fresh or frozen. For universal use QIAamp (QIAGEN) was most suitable, frozen blood was extracted best using NucleoSpin Tissue (Macherey-Nagel). PMID- 12532900 TI - [Incidence of S. typhimurium in the Czech Republic 1999-2001]. AB - The ratio of serovars of S. Typhimurium among notified cases of salmonelloses in 1999-2001 did not exceed 3% and so far has a slightly declining trend. On a long term basis the highest morbidity rate caused by this agent is reported in the South Moravian region. The age distribution and specific morbidity is highest in 1-4-year-old children, the seasonal incidence is highest, in the summer months. The most frequently suspected vehicles are eggs, poultry and meat products, in 1999 for the first time as a vehicle a grilled piglet was reported, in a total of three epidemics. Epidemiological analysis of salmonelloses caused by S. Typhimurium revealed that in the CR individual characteristics do not differ from those caused by S. Enteritidis and that the epidemiological situation as regards this serovar is, so far quiescent. PMID- 12532901 TI - [Effect of parenteral glutamine on restoration of lymphocyte subpopulations after high-dose chemotherapy and autologous hematopoietic cell transplantation: data from a double-blind randomized study]. AB - Within the framework of a randomized double blind study focused on the effect of glutamine on the clinical course of autologous transplantation of peripheral cells the authors assessed lymphocyte sub-populations (CD3, CD4, CD8, CD19 and CD57+ cells) before transplantation and 14, 28 and 42 days after transplantation. A total of 36 patients were investigated (18 glutamine, 18 placebo). In the whole group of patients the authors found restoration of CD4 and CD19 cells to pretransplantation values one day +42 after transplantation, in CD8 and CD57 cells a statistically significant increase as compared with the pre transplantation state occurred. In the glutamine group they observed on day +28 a more rapid restoration of CD8 and a marginally better restoration of CD19 positive cells, while patients who were given placebo restored CD57+ cells more rapidly. All these differences were balanced on day +42, only CD19+ cells were at that time marginally higher in the placebo group. With the exception of CD19+ lymphocytes the authors observed weak correlations between the number of lymphocytes on day +42 after transplantation and the number of transplanted CD34+ cells. It may thus be stated that the drop of lymphocyte sub-populations has a short-term character, the restoration correlates among others with the administered amount of haematopoietic cells. Significant importance of glutamine for the restoration of the lymphocyte sub-population was however not proved. PMID- 12532902 TI - [Deep necrotizing soft tissue infections. Necrotizing fasciitis and necrotizing myositis]. AB - The authors present a review on deep soft tissue infections--necrotizing fasciitis and necrotizing myositis, incl. the classification of myositis into sub groups according to the causal agent. The authors describe the incidence of these diseases, basic clinical symptoms and therapeutic principles. Therapy is based on surgical treatment, antimicrobial treatment and in indicated cases on hyperbaric oxygen therapy (HBO). PMID- 12532903 TI - [Biofilms and their significance in medical microbiology]. AB - Microorganisms are able to adhere to various surfaces and to form there a three dimensional structure known as biofilm. In biofilms, microbial cells show characteristics and behaviours different from those of plankton cells. Intercellular signalizations of the quorum-sensing type regulate interaction between members of the biofilm. Bacteria embedded in the biofilm can escape and form well known planktonic forms, that are obviously only a part of the bacterial life cycle. Bacteria adhere also to medically important surfaces such as catheters, either urinary or intravenous ones, artificial heart valves, orthopedic implants and so on and contribute to device-related infections like cystitis, catheter-related sepsis, endocarditis etc. Once a biofilm has been established on a surface, the bacteria harboured inside are less exposed to the host's immune response and less susceptible to antibiotics. As an important cause of nosocomial infections the biofilm must remain in the centre of the microbiologist's attention. PMID- 12532904 TI - [Orofacial clefts in western Slovakia]. AB - Orofacial clefts are relatively frequent inborn developmental defects. The objective of our study was to reveal the actual frequency of orofacial clefts in western Slovakia. From 390 404 liveborn children in Bratislava and the western Slovakian region in 1985-1999, 629 children with orofacial clefts were operated at a specialized department--the Clinic of Plastic and Aesthetic Surgery of the Medical Faculty, Comenius University Bratislava. The authors assessed the total incidence of orofacial clefts from the records of the above department and reported data. For statistical evaluation of differences they used the chi 2 test. The total incidence during the years of investigation in western Slovakia was 1.611/10(3) liveborn children whereby in Bratislava town the incidence was higher than in the western Slovakian region. Between individual districts these values had a wide range from 1.20-2.04/10(3) liveborn infants. As regards different types, the authors found the highest incidence of orofacial clefts of the primary and secondary palate. The frequency in males was significantly higher. As regards the risk of possible birth of an infant with this congenital developmental defect in western Slovakia, the authors found a discrepancy between data on their records--1 child/620 liveborn infants, and notified cases--1 child/756 liveborn children. PMID- 12532905 TI - [Primary prevention of malignant tumors]. AB - Primary prevention represents an effective instrument in the battle against tumour diseases. Prevention is aimed to lower the mortality for this disease by means of reduction of the cancer incidence. Our article reviews the main fields of primary prevention, namely the principles of proper nutrition and regimen. Effects of balance in energy and related nutrients, essential components of the diet, types of meals together with problems of food processing and storage are discussed from the view of the cancer prevention. Each chapter brings applicable preventive recommendations. Smoking is considered as a risks factor not only for tumours, but also for other serious diseases. Other carcinogenic factors and possible preventive measures are referred in the last chapter. Though we know that the primary cancer prevention requires active approach of the whole society, we are certain that medical professionals in every day contact with patients can highly improve the public informedness. The article submits substrate for such discussions. PMID- 12532906 TI - [Current views on the development of diabetic retinopathy]. AB - The basic pathogenic mechanism of the diabetic retinopathy is hyperglycaemia. Capillary damage could be explained by several mechanisms. Among many causes the sorbitol overload, the excessive storage of advanced glycosylation endproducts, oxidative stress and/or the cell apoptosis after the paradoxical glucose starvation can be emphasised. The article further analyses mechanism of capillary damage in the retina is on the basis of hemorheological, hemodynamical and histopathological changes. The specific role of the retinal pigment epithelium is described. Histopathological changes accompanying the elementary microcirculatory anomalies result from the apoptotic death of pericytes, damage of endothelial cells, and impairment of the basal membrane. Microcirculatory changes, which include the capillary dilation, vascular occlusion, and proliferation of new vessels, bring about further development of the diabetic retinopathy. PMID- 12532907 TI - [Evaluation of the severity of vascular impairment in the lower extremities in patients with chronic kidney failure]. AB - BACKGROUND: Arterial ischemic disease represents highly serious complication for patients undergoing chronic haemodialysis. The aim of the study was to examine peripheral arteries in the group of haemodialysed patients and to estimate the incidence and seriousness of pathological findings. METHODS AND RESULTS: Total 82 sonographic examinations of arteries in lower extremities using Multi Dopplex II set with graphic registration of dopplex curves was done in 53 dialysed patients. Blood pressure values of four peripheral arteries at legs together with blood pressure values at the brachial artery were evaluated as the ankle-arm index (API). Character of changes of dopplex curves was classified as SO index (obliteration index). Also with normal API, changes in SO were identified. SO index was significantly higher (p = 0.034) in patients of the peritoneal dialysis program in comparison with haemodialysed patients, in persons with diabetes the value was only marginally higher. Patients who died later (of the cardiovascular complications), the SO index was significantly higher then in others (p < 0.0001), though the API values were not different. Repeated testing in 29 patients after 6 months brought about worsening of the SO index (p < 0.00001) in those patients where no changes of API were seen. No relation with any of the followed biochemical parameters was found. CONCLUSIONS: To estimate most accurately the conditions of the peripheral arteries in dialysed patients ankle arm index was used together with evaluation of the shape of dopplex curves (index SO). Such detailed Doppler examination has proved identifiable changes of arteries in 73 out of 82 cases. PMID- 12532908 TI - [Invasive ductal carcinoma of the breast: study of the number of copies of the CCND1 gene and chromosome 11 using fluorescence in situ hybridization (FISH) in comparison with expression of cyclin D1 protein and estrogen receptors (ER alpha) with immunohistochemical detection]. AB - BACKGROUND: Overexpression of oncogenic proteins may be caused by gene amplifications. Cyclin D1 participates in regulation of the cell cycle. Relations between cyclin D1 expression and amplification of CCND1 gene encoding this protein in invasive duct breast carcinomas (IDC) are not fully elucidated. An increased interest is also focused on relations to the estrogen receptor (ER alpha). METHODS AND RESULTS: We investigated copy numbers of the CCND1 gene, expression of cyclin D1 and expression of ER alpha in a group of 60 females and 1 male with IDC. The age range varied from 33 to 89 years (median 57 years). The number of CCND1 gene copies and the number of chromosome 11 was evaluated using FISH, the expression of cyclin D1 and ER alpha was investigated by IHC. We detected a strong amplification of CCND1 gene (> 10 copies per tumor cell nuclei) in 9 patients, weak amplification (< or = copies) in 16 patients. Amplification of the CCND1 gene correlated well with the overexpression of cyclin D1. We observed the overexpression of cyclin D1 also in 13 of 36 patients without the gene amplification; therefore, the mechanism of the protein overexpression is different than that caused by the gene amplification in a proportion of patients. Amplification of the CCND1 gene was associated with a high histologic grade of IDC, whereas cases with cyclin D1 overexpression only were not. 24 of 31 patients with overexpression of cyclin D1 coexpressed ER alpha. We did not find correlation between expression/amplification of cyclin D1/CCND1 gene and the size of carcinomas and with metastases to the axillary lymph nodes. CONCLUSIONS: Amplification of the CCND1 gene is associated with overexpression of cyclin D1 in a majority of IDC. Overexpression of cyclin D1 is related to an increased expression of ER alpha. Interaction between cyclin D1 and ER alpha may explain low response to anti-estrogen therapy of some patients. PMID- 12532909 TI - [Cutaneous complications in idiopathic inflammatory bowel disease]. AB - A 29-year-old male patient with the anamnesis of inflammatory bowel disease and Grave-Basedowov disease was hospitalized because of rapidly spreading skin defect with affected muscle on the left shin. This skin defect appeared after the significant decreasing of corticoids. The small skin trauma preceded the pyoderma gangrenosum. First the skin disease was not right diagnosed and patient was cured by the excision of the defect. It caused tissue disintegration, muscle necrosis and extension of the defect. The whole leg was endangered. Patient was cured with corticoids and cyclosporin A after the right diagnosis. The defect healed and laboratory inflammatory markers decreased. The immunosuppresive therapy was changed to azathioprin, the corticoid therapy was interrupted. After three months the defect was healed. PMID- 12532910 TI - [The laws of the "real world" apply even to publications]. PMID- 12532911 TI - The Barcelona conference and the HIV prevention backlash. PMID- 12532912 TI - Barcelona reports on people with HIV emphasize treatment issues. PMID- 12532913 TI - Many epidemics, two causes: the virus and our inertia. PMID- 12532914 TI - Bypass without missing a beat. PMID- 12532915 TI - Taking a bite out of gum disease. PMID- 12532916 TI - Organ and tissue transplants: are they safe? PMID- 12532917 TI - What to do when prostate cancer returns. PMID- 12532918 TI - The neurosurgeon in sport: awareness of the risks of heatstroke and dietary supplements. PMID- 12532919 TI - The Forge of Vulcan, by Jacopo Tintoretto (1519-1514). PMID- 12532920 TI - Surgical anatomy of the nose in the elderly: value of conservative rhinoplasty by transoral route. AB - Functional or aesthetic rhinoplasty is being done later and later in life. It is essential to know the characteristics of morphological ageing of the nose in order to modify surgical techniques. An anatomical study was carried out on 40 fresh cadavers more than 70 years old. All the subjects studied were measured to define the external morphology of the nose. Amongst these 40 subjects, 20 were dissected plane by plane, 10 were operated on by a new conservative rhinoplasty technique utilizing a strictly transoral route and 10 were operated on by a classical rhinoplasty technique. The anatomy of the nose in the elderly subject is characterized by thinner skin, subcutaneous fat infiltrating in the muscles in 50% of cases and the small muscles of the tip of the nose undergoing complete involution. The technique of rhinoplasty described here shows the value of better respecting lesions at the tip of the nose which is particularly fragile in the aged subject. It may be extended to post-traumatic rhinoplasties or to the nose which has already been operated on. PMID- 12532921 TI - Promoting the health of student athletes. PMID- 12532922 TI - Wegener's granulomatosis: ischemic stroke as the first clinical manifestation (case study). PMID- 12532923 TI - Do randomised clinical trials influence practice? The example of cerebral vein and dural sinus thrombosis. PMID- 12532924 TI - Paraneoplastic brainstem encephalitis and anti-Ri antibodies. PMID- 12532925 TI - Hallervorden Spatz syndrome (pantothenate kinase associated neurodegeneration) in two Sardinian brother with homozygous mutation in PANK 2 gene. PMID- 12532926 TI - Electrophysiological evidence of trigeminal neuropathy in pseudotumor cerebri. PMID- 12532927 TI - Whole-brain T1-relaxation time measurements in multiple sclerosis. PMID- 12532928 TI - Normal d-dimer levels do not exclude the diagnosis of cerebral venous sinus thrombosis. PMID- 12532929 TI - Neuralgic amyotrophy as a manifestation of infectious mononucleosis. PMID- 12532930 TI - A novel insert mutation in gamma-sarcoglycan gene leads to severe childhood autosomal recessive muscular dystrophy. PMID- 12532931 TI - Granulomatous myositis associated with antithyroglobulin and antithyroid peroxidase antibodies: an analogy to Hashimoto's encephalitis? PMID- 12532932 TI - Good cognitive outcome of patients with herpes zoster encephalitis: a follow-up study. PMID- 12532933 TI - Myasthenia gravis with diffuse alopecia areata and pemphigus foliaceus. PMID- 12532934 TI - Paroxysmal exercise-induced dystonia associated with hypoglycaemia induced by an insulinoma. PMID- 12532935 TI - Spastic tetraparesis in a patient with pseudopseudohypoparathyroidism. PMID- 12532936 TI - Acute transverse myelitis associated with coxiella burnetii infection. PMID- 12532937 TI - Seroprevalence of Borna disease virus antibodies is not increased in patients with amyotrophic lateral sclerosis. PMID- 12532938 TI - Occipital neuralgia as isolated symptom of an upper cervical cavernous angioma. PMID- 12532939 TI - The Primrose syndrome with progressive neurological involvement and cerebral calcification. PMID- 12532940 TI - Chronic acquired polyneuropathy in infancy. PMID- 12532941 TI - Acute myelitis in early Borrelia burgdorferi infection. PMID- 12532942 TI - Internal ophthalmoplegia with anti-GQ1b IgG antibody. PMID- 12532943 TI - Mental retardation, peculiar facial dysmorphism and HbH inclusions in a 4-year old boy. PMID- 12532944 TI - Thalamo-mesencephalic infarction as a single manifestation of combined factor V Leiden and antiphospholipid syndrome. PMID- 12532945 TI - Long-term virological response and growth rate of children with chronic hepatitis C who received natural interferon-alpha. PMID- 12532946 TI - Treading water: Americans' access to needed medical care, 1997-2001. AB - Despite unprecedented economic growth, low unemployment and fewer uninsured people, Americans' ability to get needed medical care failed to improve significantly between 1997 and 2001, according to findings from the Center for Studying Health System Change's HSC) Community Tracking Study Household Survey. While most people get the care they believe they need, about one in seven Americans reported some difficulty obtaining needed care in 2001 - about the same as in 1997. At the same time, health system-related problems - such as the ability to get timely appointments - increased, suggesting possible health system capacity constraints are emerging. On a brighter note, children's ability to get needed care improved. PMID- 12532947 TI - A patient with Kawasaki disease who developed acute urinary retention due to pelvic neuroblastoma. PMID- 12532948 TI - Life-threatening intoxication in twin neonates following accidental overdose of rectal dimenhydrinate. PMID- 12532949 TI - Increased risk of multicystic dysplastic kidney among babies of both pre gestational and gestational diabetic mothers. PMID- 12532950 TI - Endothelin and cardiac development. PMID- 12532951 TI - The physician's role in helping patients to increase physical activity and improve eating habits. PMID- 12532952 TI - Thrombolysis for stroke: defining the time window. PMID- 12532953 TI - Genetic association studies of stroke: hope, signal, and noise. PMID- 12532954 TI - Amaurosis in carotid stenosis versus atrial fibrillation: prediction or observation? PMID- 12532955 TI - The insurance gap and minority health care, 1997-2001. AB - Gaps in access to medical care among working-age white Americans, African Americans and Latinos failed to improve between 1997 and 2001, despite a booming economy and increased national attention to narrowing and eliminating minority health disparities. African Americans and Latinos continue to have less access to a regular health care provider, see a doctor less often and lag behind whites in seeing specialists, according to recent findings from the Center for Studying Health System Change (HSC). Ethnic and racial disparities in access among uninsured Americans are much greater than disparities among the insured. Uninsured whites' greater financial resources may explain why they have fewer problems accessing care. Eliminating disparities in minority health care will be difficult without first eliminating these gaps in minority health insurance. PMID- 12532956 TI - Does the weather influence stroke incidence? PMID- 12532957 TI - Computed tomography of hyperacute infarctions in relation to intra-arterial clot lysis. PMID- 12532958 TI - Trends in stroke mortality: the impact of the Year 2000 Age Standard and the International Statistical Classification of Diseases and Related Health Problems, 10th Revision. PMID- 12532959 TI - Reliability of angiographic assessment of brain arteriovenous malformations. PMID- 12532960 TI - From the Centers for Disease Control and Prevention. Exophiala infection from contaminated injectable steroids prepared by a compounding pharmacy--United States, July-November 2002. PMID- 12532961 TI - Beyond hypertension: unraveling the causes of intracerebral hemorrhage. PMID- 12532962 TI - From the Centers for Disease Control and Prevention. Provisional surveillance summary of the West Nile virus epidemic--United States, January-November 2002. PMID- 12532963 TI - Folate and the risk of stroke: fortify first and ask questions later? PMID- 12532964 TI - From the Centers for Disease Control and Prevention. Intrauterine West Nile virus infection--New York, 2002. PMID- 12532965 TI - From the Centers for Disease Control and Prevention. Additions to terrorism preparedness compendium. PMID- 12532966 TI - Assessment of regional acute stroke care. PMID- 12532967 TI - Thrombolysis and hemorrhagic transformation. PMID- 12532968 TI - Who do you trust? Americans' perspectives on health care, 1997-2001. AB - A turbulent backlash against managed care in the mid-1990s pitted consumers and health care providers against health plans in a struggle for control over medical decision making. New findings from the Center for Studying Health System Change (HSC) Community Tracking Study Household Survey indicate consumer confidence in the system and trust in physicians increased slightly between 1997 and 2001, perhaps as a result of changes in laws and loosening of health plan restrictions. Nevertheless, there is strong evidence of continued public concern about the influence of health plans on medical decision making. For example, the level of trust in their physicians among people in fair or poor health has not increased, and more than 40 percent of privately insured Americans continue to believe their doctor is strongly influenced by health plan rules when deciding about their care. PMID- 12532969 TI - Working families' health insurance coverage, 1997-2001. AB - Despite a booming U.S. economy, falling unemployment and moderate health insurance premium growth, the percentage of working Americans and their families with employer-sponsored health insurance failed to increase substantially between 1997 and 2001, according to findings from the Center for Studying Health System Change (HSC) Community Tracking Study Household Survey. There were, however, dramatic changes in the insurance status of people who lacked access to or did not take up employer coverage: fewer uninsured, more public program enrollment and a decline in coverage by individual insurance and other sources. While the State Children's Health Insurance Program (SCHIP) clearly reduced uninsurance among low-income children, evidence also suggests a fair amount of substitution of public insurance for private coverage. PMID- 12532970 TI - WHO report on violence and health. PMID- 12532971 TI - IOM: lack of health insurance puts families at risk. PMID- 12532972 TI - Kinder and gentler: physicians and managed care, 1997-2001. AB - Despite the managed care backlash, an overwhelming majority of U.S. physicians continue to contract with managed care health plans. In fact, according to a new Center for Studying Health System Change (HSC) study, between 1997 and 2001 physicians reported a modest increase in the proportion of practice revenue from managed care contracts and the average number of contracts. At the same time, the nature of physicians' relationships with health plans changed, with a significant decrease in plans' use of capitation, or fixed monthly payments for each patient regardless of the amount of care provided. Meanwhile, physician practices moved away from using direct financial incentives to influence doctors' clinical decision making, but did experience an increase in the overall influence of treatment guidelines and other practices commonly associated with managed care. PMID- 12532973 TI - Mounting pressures: physicians serving Medicaid patients and the uninsured, 1997 2001. AB - The proportion of doctors providing any charity care decreased from 76.3 percent in 1997 to 71.5 percent in 2001, according to a new study by the Center for Studying Health System Change (HSC). The proportion of physicians serving Medicaid patients also decreased from 87.1 percent in 1997 to 85.4 percent in 2001. The small decrease in physicians serving Medicaid patients does not appear to have had any negative effects on access to physicians among Medicaid beneficiaries. On the other hand, the more sizable decrease in physicians providing charity care is consistent with other evidence showing decreased access to physicians by uninsured persons. New budget pressures could lead states to freeze or cut Medicaid provider payment rates, which could then trigger access problems. PMID- 12532974 TI - JAMA patient page. Lung cancer. PMID- 12532975 TI - Ultrasonography of temporal arteries discloses temporal arteritis in "pure" polymyalgia rheumatica patients. PMID- 12532976 TI - Cloning claim is science fiction, not science. PMID- 12532977 TI - Graduate students' views on unionization. PMID- 12532978 TI - Graduate students' views on unionization. PMID- 12532979 TI - International Conference on Harmonisation: guidance on Q1D bracketing and matrixing designs for stability testing of new drug substances and products; availability. Notice. AB - The Food and Drug Administration (FDA) is announcing the availability of a guidance entitled "Q1D Bracketing and Matrixing Designs for Stability Testing of New Drug Substances and Products." The guidance was prepared under the auspices of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). This guidance is an annex to an ICH guidance entitled "Q1A(R) Stability Testing of New Drug Substances and Products" (66 FR 56332, November 7, 2001). It is intended to provide guidance on the application of reduced designs (i.e., bracketing and matrixing) for stability studies conducted in accordance with the principles outlined in ICH Q1A(R). PMID- 12532980 TI - An information sheet for patient. Questions and answers about drugs used to treat and prevent influenza. PMID- 12532981 TI - A universal view of data. Interview by Anne Zender. PMID- 12532982 TI - Critiquing unpublished theories. PMID- 12532983 TI - Resources for affordable prescription medicine. PMID- 12532984 TI - [STS-95 space experiments (plants and cell biology)]. PMID- 12532985 TI - [Gravimorphogenesis of Cucurbitaceae seedlings: negatively controlled morphogenesis by gravity and its mechanism]. PMID- 12532986 TI - [Plant growth and development, and auxin polar transport in space conditions]. PMID- 12532987 TI - [Growth regulation mechanisms in higher plants under microgravity conditions]. PMID- 12532988 TI - [Effects of gravity on growth of testis and spermatogenesis in mice]. PMID- 12532989 TI - [The change of human sperm motility in parabolic flight]. PMID- 12532990 TI - [Effect of the clinostat rotation on blastokinesis of the silkworm, Bombyx mori]. PMID- 12532991 TI - [Gravity effect on the kinematics and differentiation tendency of the A6 cells]. PMID- 12532992 TI - [Effects of gravity on the gene expression of osteoclast differentiation factor and osteoclastogenesis inhibitory factor]. PMID- 12532993 TI - [Analysis of surface phenotype of human peripheral blood lymphocyte after exposure to gravity stress]. PMID- 12532994 TI - Science background and strategic planning of space chronobiology. PMID- 12532995 TI - [Effects of spaceflight and/or hindlimb suspension on rat neck muscle]. PMID- 12532996 TI - [Analysis of eye torsion in fish evoked by linear acceleration]. PMID- 12532997 TI - [Effects of postural changes on nystagmus elicitation induced by lateral linear acceleration]. PMID- 12532998 TI - [Development of medaka embryo under simulated microgravity by 3D-clinostat: development of retina and cartilage]. PMID- 12532999 TI - [Changes in visual acuity during development of medaka larvae and their behavior under microgravity]. PMID- 12533000 TI - [Microgravity behavior of medaka fish with or without otolith information]. PMID- 12533001 TI - [Posture analysis in the rat exposed to microgravity during parabolic flight]. PMID- 12533002 TI - [Activities, which modulate mutability of cells, in human serum after parabolic flight]. PMID- 12533003 TI - [Effects of microgravity on development of the baroreflex mechanisms--preliminary reports of Neurolab program]. PMID- 12533004 TI - [STS-95 neural activity measuring experiment]. PMID- 12533005 TI - [VFEU water quality control in STS-95 aquatic animal experiment]. PMID- 12533006 TI - [Separation of plant cells in size from 10 to 15 micrometers and gelation of gellangum solution containing plant cells under microgravity]. PMID- 12533007 TI - [Protein crystal growth and influences of microgravity]. PMID- 12533008 TI - [Development of plant growth chambers for the experiments under microgravity conditions (4)-results of two experiments for water circulation in parabolic flight]. PMID- 12533009 TI - [Development of plant growth chambers for the experiments under microgravity conditions--development of measurement system of plant water uptake]. PMID- 12533010 TI - [Development of plant growth chambers for the experiments under microgravity conditions (7) -measurements of leaf temperature and net photosynthetic rates of leaves in a parabolic airplane flight experiment]. PMID- 12533011 TI - [SOS response in X-ray- or UV-irradiated E. coli cells under microgravity]. PMID- 12533012 TI - [The effect of microgravity on induced mutation through DNA synthesis during space flight]. PMID- 12533013 TI - [STS-95 space experiment: analysis of mutations induced in human tumor cells]. PMID- 12533014 TI - [Space radiation environment around International Space Station]. PMID- 12533015 TI - [Growth promotion and cell wall changes in rice coleoptiles under microgravity conditions]. PMID- 12533016 TI - [Growth and cell wall metabolism in azuki bean epicotyls change reversibly according to the magnitude of gravity]. PMID- 12533017 TI - [Stimulation of growth and xyloglucan breakdown in Arabidopsis hypocotyls under microgravity conditions]. PMID- 12533018 TI - [Automorphogenesis of rice and Arabidopsis seedlings under microgravity conditions]. PMID- 12533019 TI - [Effects of various gravistimuli on senescence of oat leaf segments]. PMID- 12533020 TI - [Microtubule reorganization in response to gravistimulation during cucumber peg development]. PMID- 12533021 TI - Appendiceal colic: A rare complication of colonoscopy. AB - We report a case of appendiceal colic presenting as chronic right lower quadrant abdominal pain, without leukocytosis or other inflammatory signs, following an uneventful colonoscopic exam without biopsy or polypectomy. The resected appendix appeared grossly abnormal and was engorged with fecal material. The patient was immediately relieved of pain and remains so four years postoperatively. We conclude that insufflation of the cecum caused inspissation of fecal material into the lumen of the appendix. The ensuing incomplete or partial obstruction of the appendiceal lumen resulted in development of appendiceal colic without any signs or symptoms of inflammation. PMID- 12533022 TI - Uncommitted Xenopus blastula cells can be directed to uniform muscle gene expression by gradient interpretation and a community effect. AB - The animal cap cells of Xenopus blastulae behave as multi-potent stem cells in so far as they can differentiate along many unrelated pathways according to the kind and amount of signal factor that they experience. At first, animal cap cells activate early zygotic genes across a broad range of TGFbeta concentrations; soon after this, they activate later genes more intensely and over a narrow concentration range. Here we show that uncommitted blastula cells can be directed, by the sequential influence of a particular concentration of a TGFbeta morphogen and an FGF-mediated community effect, to form a homogeneous single cell type. As a result of these two signalling processes, an entire population of animal cap cells can be converted, in the absence of other signals, to a uniform population of one tissue type. Mesoderm cells that experience a particular concentration of activin increase their XMyoD expression by 10-fold and become distinct from neighbouring cells that received lower or higher concentrations of activin. The signalling processes that we employ here may be important in normal development and useful in guiding stem cell differentiation. PMID- 12533023 TI - The circadian gene Clock is required for the correct early expression of the head specific gene Otx2. AB - The circadian cycle is a universal molecular mechanism for imposing cyclical control on cellular processes. Here we have examined the role of one of the crucial circadian genes, Clock, in early Xenopus development. We show that a dominant negative version of Clock can block the function of the endogenous Clock gene. Doing so during early development reduces Otx2 expression in a highly specific manner and results in anterior defects. Together with previous work (Green et al. (2001) Mech. Dev. 105-110), these results suggest that a positive regulatory loop exists between Clock and Otx2. PMID- 12533024 TI - Origin of coronary endothelial cells from epicardial mesothelium in avian embryos. AB - It has been established that coronary vessels develop through self-assembly of mesenchymal vascular progenitors in the subepicardium. Mesenchymal precursors of vascular smooth muscle cells and fibroblasts are known to originate from an epithelial-to-mesenchymal transformation of the epicardial mesothelium, but the origin of the coronary endothelium is still obscure. We herein report that at least part of the population of the precursors of the coronary endothelium are epicardially-derived cells (EPDCs). We have performed an EPDC lineage study through retroviral and fluorescent labelling of the proepicardial and epicardial mesothelium of avian embryos. In all the experiments onlythe surface mesothelium was labelled after 3 h of reincubation. However, endothelial cells from subepicardial vessels were labelled after 24-48 h and endothelial cells of intramyocardial vessels were also labelled after 48-96 h of reincubation. In addition, the development of the coronary vessels was studied in quail-chick chimeras, obtaining results which also support a mesothelial origin for endothelial and smooth muscle cells. Finally, quail proepicardial explants cultured on Matrigel showed colocalization of cytokeratin and QH1 (mesothelial and endothelial markers, respectively) after 24 h. These results, taken together, suggest that EPDC show similar competence to that displayed by bipotential vascular progenitor cells [Yamashita et al., Nature 408: 92-96 (2000)] which are able to differentiate into endothelium or smooth muscle depending on their exposure to VEGF or PDGF-BB. It is conceivable that the earliest EPDC differentiate into endothelial cells in response to myocardially-secreted VEGF, while further EPDC would be recruited by the nascent capillaries via PDGFR-beta signalling, giving rise to mural cells. PMID- 12533025 TI - KL/KIT co-expression in mouse fetal oocytes. AB - The tyrosine kinase receptor, KIT, and its ligand, KL are important regulators of germ cell development. The aim of this study was to examine in detail the expression of the genes encoding these proteins (White and Steel, respectively) during the fetal period (14.5-18.5 days post coitum, dpc) and the two weeks after birth in mouse ovaries using the highly sensitive in situ reverse-transcriptase polymerase chain reaction (in situ RT-PCR). KL and KIT mRNAs were not detected in 14.5-15.5 dpc ovaries but, between 16.5 and 17.5 dpc, most of the oocytes in the outer regions of the ovaries positively stained for both mRNAs. The majority of the co-expressing oocytes were identified at the zygotene/pachytene stage of meiotic prophase I. At 18.5 dpc, positive staining for KL mRNA was present only in the somatic cells in the outer regions of the ovaries. At birth, faint KL mRNA labelled somatic cells were mainly found in the central region of the ovaries and, by P7-14, a higher level of expression was detected in the follicle cells of one- and two-layered growing follicles. Between 17.5 dpc and birth, most of the oocytes expressed KIT mRNA and, from P7 onward, there was a considerable accumulation of transcripts in the growing oocytes. The results of in situ RT-PCR were confirmed by RT-PCR on purified populations of oocytes, and at protein level by means of immunohistochemistry. The co-expression of KL and KIT in a fraction of fetal oocytes suggests that the KL/KIT system, besides the well known paracrine functions on germ cells, may exert a novel autocrine role during the mid-stage of the oocyte meiotic prophase. The possibility that this autocrine loop plays a role in sustaining the survival of fetal oocytes in this stage is supported by the finding that the addition to the culture medium of anti-KL or anti-KIT antibodies led to a significant increase in oocyte apoptosis in the absence of exogenous KL. PMID- 12533026 TI - Autoaggressive, multi-headed and other mutant phenotypes in Hydractinia echinata (Cnidaria: Hydrozoa). AB - In an inbreeding program conducted with the colonial hydroid Hydractinia echinata, each F1 mating produced up to 50% F2 offspring displaying an aberrant, clone-constant phenotype, hence referred to as mutant strain. In autoaggressive strains, in one or several areas of the colony autoreactive stolons direct their aggressive devices (stolon tips filled with cytotoxic stinging cells), normally used to kill allogeneic competitors for living space, towards neighboring stolons or polyps (hydranths) of their own colony. In these areas tumor-like masses of self-aggressive stolons were formed, in severe cases causing the death of the colony. Based on previous genetic studies, the interpretation proposed here attributes autoaggressive behavior to a mosaic-type alternative expression of arl (allorecognition) alleles in heterozygous individuals. Developmental mutant strains termed He-mh form supernumerary heads during regeneration and normal development as well. Common to all He-mh phenotypes isthe production of additional headsalong the bodycolumn of fully-grown polyps. The heads give rise to complete hydranths connected by a tube that derives from the gastric region of the original polyp and eventually transforms into a stolon. In bastol strains, polyps convert the basal region of their body column into a periderm-covered stolon from which the residual apical hydranth detaches. Colonies expressing both the He-mh and the bastol (bst) phenotype frequently lose detaching multi-headed hydranths and the colony disintegrates. The large number of mutant F2 offspring reveals high genetic variability in Hydractinia. PMID- 12533027 TI - Young microspore-derived maize embryos show two domains with defined features also present in zygotic embryogenesis. AB - In this work we report the existence of two domains in early microspore maize proembryos displaying similar features of zygotic embryogenesis. The large or so called endosperm-like domain exhibits specific features: coenocytic organisation, synchronous mitosis, vacuolated cytoplasm, starch granules, incomplete walls containing callose and differential tubuline organisation. The small or embryo like domain displays small polygonal uninucleate cells with typical organisation of proliferating cells. The structural organisation and the subcellular localization of specific cytoplasmic and cell wall components (starch, tubuline and callose) in both proembryo domains have been determined by using specific cytochemical and immunocytochemical methods. Four morphological types of proembryos containing both domains have been characterised. Taking into account their relative size, the high asynchrony of the culture and the homologies between structural features of endosperm-like domain and zygotic endosperm development, they could represent different stages in microspore embryogenesis development. The ZmAE1 and ZmAE3 genes expressed in the Embryo Surrounding Region of the endosperm during zygotic embryogenesis (Magnard et al., 2000), were revealed to be expressed in early microspore proembryos by in situ hybridization at light and electron microscopy levels. This data supports the existence of an endosperm-like function during early microspore embryogenesis and provides new insights into the onset of microspore embryogenesis in maize, and its parallelism with zygotic embryogenesis. PMID- 12533028 TI - Quantitation of 3D ureteric branching morphogenesis in cultured embryonic mouse kidney. AB - The growth and branching of the epithelial ureteric tree is critical for development of the permanent kidney (metanephros). Current methods of analysis of ureteric branching are mostly qualitative. We have developed a method for measuring the length of individual branches, and thereby the total length of the ureteric tree in 3 dimensions (3D). The method involves confocal microscopy of whole-mount immunostained metanephroi and computer-based image segmentation, skeletonisation and measurement. The algorithm performs semi-automatic segmentation of a set of confocal images and skeletonisation of the resulting binary object. Length measurements and number of branch points are automatically obtained. The final representation can be reconstructed providing a fully rotating 3D perspective of the skeletonised tree. After 36 h culture of E12 mouse metanephroi, the total length of the ureteric tree was 6103 +/- 291 microm (mean +/- SD), a four-fold increase compared with metanephroi cultured for just 6 h (1522 +/- 149 microm). Ureteric duct length increased at a rate of 153 microm/h over the first 30 h period and was maximal between 18 and 24 h at 325 microm/h. The distribution of branch lengths at the six time points studied was similar, suggesting tight control of ureteric lengthening and branching. This method will be of use in analysing ureteric growth in kidneys cultured in the presence of specific molecules suspected of regulating ureteric growth. The method can also be used to analyse in vivo kidneys and to quantify branching morphogenesis in other developing organs. PMID- 12533029 TI - Cysteine-rich region of X-Serrate-1 is required for activation of Notch signaling in Xenopus primary neurogenesis. AB - The Notch family genes encode single-pass transmembrane proteins which function in a variety of cell fate specifications in invertebrates and vertebrates. In Xenopus primary neurogenesis, the Notch ligands, X-Delta-1 and X-Serrate-1, mediate Notch signaling and regulate cell differentiation. In the present study, we examined the role of the Serrate-specific cysteine-rich (CR) region in the primary neurogenesis of Xenopus embryos. The ligand constructs containing the DSL (Delta/Serrate/Lag-2) domain in the extracellular region caused a reduction in primary neurons, whereas the DSL-deleted form of X-Delta-1 resulted in the overproduction of primary neurons. However, the DSL-deleted form of X-Serrate-1 or the construct containing only the CR region in the extracellular domain (SerCR) reduced the number of primary neurons. In contrast, the CR-deleted form of X-Serrate-1 (SerACR) lost activity as a Notch ligand, regardless of the presence of the DSL domain within the extracellular domain. Overexpression of X Delta-1 and X-Serrate-1 strongly induced the expression of Xenopus ESR-1 (XESR 1), a gene related to Drosophila Enhancer of split. SerCR alone also moderately induced the expression of XESR-1, but the SerACR form did not induce this expression. Co-injection of X-Notch-1deltaICD, which deletes the intracellular domain (ICD), with SerCR suppressed a neurogenic phenotype, although co-injection of X-Su(H)1DBM with SerCR did not, indicating that SerCR affects primary neurogenesis through the Notch/Su(H) pathway. These results suggestthatthe CR region of Xenopus Serrate is required for the activation of Notch signaling and cell fate specification in primary neurogenesis. PMID- 12533030 TI - Lymphocyte function-associated antigen-1 and intercellular adhesion molecule-1 expression on B-cell subsets and the effects of splenectomy-experimental studies. AB - There is an abundance of data dealing with recirculation of T cells in the rats, but relatively little is known about the traffic of B cells. The adhesion molecules expressed on the surface membrane are of great significance for recirculation of lymphocytes. However, very little is known about the expression of various adhesion molecules on B-cell subsets. Here we show that in normal rats various adhesion molecules are differentially expressed on B-cell subsets and that the level of their expression changes after the entry of B lymphocytes from the blood into the lymphoid tissues. In splenectomized rats, the surface expression of LFA-1 and ICAM-1 is selectively reduced on B-cell subsets in blood and lymph node, which is accompanied by a selective increase in the number of all B-cell subsets in the blood. The decreased surface expression of adhesion molecules results in faster migration of B lymphocytes through lymph nodes with subsequent accumulation of these cells in the blood. PMID- 12533031 TI - The combination of chemotherapy and systemic immunotherapy and the concept of cure in murine leukemia and lymphoma. AB - While important advances have been made in our understanding of the molecular and biochemical processes that lead to the malignant transformation of myeloid and lymphoid cells, no major breakthroughs leading to long-term survival of patients of acute myeloid leukemia (AML) and lymphoma have been achieved during the last decade. Treatment failure, particularly in AML, is mostly related to the problem of resistance to multiple chemotherapeutic drugs and the morbidity and mortality associated with intensive chemotherapy. Thus, a significant challenge that remains is to develop novel therapeutic strategies that would ideally be able to address these issues. Novel immune approaches to cancer immunotherapy, while promising for specificity and long-term protection as single therapies, have not typically proven potent enough to generate long-lasting therapeutic responses. Recent evidence suggests that combining immunotherapy with chemotherapy can lead to increased effectiveness of chemotherapy without making the treatment intolerable to patients. This review focuses on the role of T cell costimulation in tumor immunosurveillance and on the therapeutic efficacy of a combination regimen consisting of chemotherapy and immunotherapy with recombinant B7.2-IgG fusion protein in preclinical AML and lymphoma models. PMID- 12533032 TI - Autoimmune phenomena in patients with myelodysplastic syndromes and chronic myelomonocytic leukemia. AB - Autoimmune paraneoplastic syndromes are commonly encountered in patients with myelodysplastic syndromes (MDS). A review of case reports and small series suggest as many as 10% of MDS patients may experience various autoimmune syndromes. Clinical manifestations of such phenomena may include an acute systemic vasculitic syndrome, skin vasculitis, fever, arthritis, pulmonary infiltrates, peripheral polyneuropathy, inflammatory bowel disease, glomerulonephritis, and even classical connective tissue disorders, such as relapsing polychondritis. On the other hand, asymptomatic immunologic abnormalities have also been reported in these patients. These autoimmune manifestations frequently respond to immunosuppressive agents including steroids and occasional hematologic responses to steroid therapy have also been reported. We report five patients with history of MDS who manifested different spectrums of autoimmune phenomena including: pyoderma gangrenosum (PG), vasculitis, Coombs negative hemolytic anemia, idiopathic thrombocytopenia, and chronic inflammatory demyelinating polyneuropathy (CIDP). We also review the incidence, nature, course and response to therapy of these manifestations and discuss potential pathogenic mechanisms. PMID- 12533033 TI - Total skin electron irradiation in mycosis fungoides: comparison between a modified Christie Hospital translational technique and the Stanford technique. AB - Seventy-one patients with mycosis fungoides (MF) were treated by Total skin electron irradiation (TSEI) using either a modified Christie Hospital translational technique (44 pts) or a six dual-field Stanford technique (27 pts). There was no statistical difference in response rate, disease-free survival and overall survival between the two irradiation techniques. However, the Stanford technique was significantly less toxic than the modified Christie Hospital technique. PMID- 12533034 TI - Veno-occlusive disease of the liver after blood and marrow transplantation: analysis of pre- and post-transplant risk factors associated with severity and results of therapy with tissue plasminogen activator. AB - We reviewed our blood and marrow transplantation (BMT) database from April 1982 to July 1996 and identified 111 of 474 patients with serum bilirubin concentration (SBR) > or = 34 micromol/l for two consecutive days within the first 20 days after related allogeneic or autologous BMT. Of the 111, 73 fulfilled the Seattle criteria for veno-occlusive disease of the liver (VOD) and had no other obvious cause for liver dysfunction. The patients were 16-60 years old (median, 39 years), and 41 were male (56%). Fourteen patients (19%) had autologous BMT, and 59 (81%) had allogeneic BMT. Twenty-eight (38%), 12 (16%), and 33 (45%) patients had severe, moderate, and mild VOD, respectively, by Seattle criteria. None of 23 patients with maximum (max) SBR > or = 257 micromol/l survived, all patients with max SBR < or = 128 micromol/l survived, and 7 of 15 patients (47%) with max SBR 128-257 micromol/l survived. The only pre transplantation risk factor predictive of severe VOD was advanced disease state (P = 0.035), and the only transplant factors that predicted severe VOD were max SBR (P = 0.01) and maximum blood urea level (P = 0.03). Ten patients (all with creatinine levels > or = 150 micromol/l) were treated with tissue plasminogen activator; only two had a significant response and only one survived beyond day 120. PMID- 12533035 TI - Primary cutaneous follicle center cell lymphoma and limited stage follicular non Hodgkin's lymphoma: a comparison of clinical and biological features. AB - Primary cutaneous follicular lymphoma (PCFL) and nodal follicular lymphoma (NFL) are different entities, which, nevertheless, exhibit common features. The aim of this study was to compare the clinico-biological characteristics and the outcome of patients with PCFL and with limited stage NFL. A group of 22 consecutive patients with PCFL presenting with single or multiple cutaneous lesions was compared to a group of 21 patients with limited stage NFL. The median age was 56 and 55 years, respectively. The histologic features were compared, as well as treatment modalities and response. Treatment of PCFL consisted of restricted field radiotherapy (RT), chemotherapy (CHT) and combined modalities (CM) in 12, 5 and 5 cases, respectively. Among the 21 patients with NFL, RT was employed in 13, CHT in 7 and the CM in one patient. The response to treatment was: 17 complete responses, CR (12 RT, 2 CHT, 3 CM), 3 partial responses, PR (1 CHT, 2 CM) and 2 non-responses, NR (2 CHT) in the PCFL group, while in the NFL group 18 patients attained CR (13 RT, 4 CHT, 1 CM) and 3 PR (3 CHT). The relapse rate was 22.7 and 28.5% in PCFL and NFL, respectively (median follow up of 48 and 50 months). The estimated 4 years event free survival (EFS) in the PCFL and NFL patients were 68 and 70%, respectively, (P = 0.83). The estimated 4 years overall survival (OS) in the PCFL and NFL groups were 90% and 100% respectively, (P = 0.254). Among the 22 patients with PCFL, there was a higher incidence of large cells than in NFL, although the differences were not significant (P = 0.08). In conclusion, despite histologic (higher proportion of large cells) and biologic differences, cutaneous and limited stage NFL show similar responses to treatment, with similar relapse rates, EFS and OS. PMID- 12533036 TI - Dose-intensified CHOP (double-CHOP) followed by consolidation with high-dose chemotherapy for high and high-intermediate risk aggressive non-Hodgkin's lymphomas. AB - Patients with aggressive non-Hodgkin's lymphoma (NHL) showing several risk factors have a poor prognosis. In such patients, conventional chemotherapy results in a low complete response rate and a high relapse rate. We performed a prospective trial of intensive induction chemotherapy followed by high-dose consolidation therapy to determine the feasibility of such an approach. We treated 39 patients with aggressive poor risk NHL with double-CHOP (D-CHOP) chemotherapy followed by high-dose therapy (HDT) with or without peripheral blood stem cell transfusion (PBSCT). The median age of the patients was 54 years (range 17-65). Twenty-seven were considered to be at high (H) and 12 were at high intermediate (H-I) risk according to the age-adjusted International Prognostic Index. Thirty-four patients (87%) responded (complete response: 74%, partial response: 13%) to D-CHOP chemotherapy. Of the 34 responders, 24 received HDT followed by PBSCT, 7 received high-dose methotrexate, and 3 refused consolidation therapy. At a median follow-up period of 26 months, the estimated 3-year overall survival rate and event-free survival rate was 64 and 51%, respectively. Our data suggest that dose-intensified D-CHOP followed by consolidation HDT is safe and appears efficacious in aggressive NHL patients with H-I and H risk. PMID- 12533037 TI - 188Re-labeled anti-CD66 monoclonal antibody in stem cell transplantation for patients with high-risk acute myeloid leukemia. AB - We have intensified the conditioning regimen prior to stem cell transplantation in 57 patients with high-risk AML and MDS by treating patients with a 188Re labeled anti-CD66 monoclonal antibody. Dosimetry was performed prior to therapy and a favorable dosimetry was observed in all cases. Radioimmunotherapy with the labeled antibody provided a mean of 15.5 Gy of additional radiation to the marrow, the kidney was the normal organ receiving the highest dose of supplemental radiation (mean 7.4 Gy): Radioimmunotherapy was followed by standard full-dose conditioning with total body irradiation (12 Gy) (n = 30) or busulfan (n = 27) and high-dose cyclophosphamide +/- thiotepa. Patients subsequently received a T cell depleted allogeneic graft from a HLA-compatible family donor (n = 24), a matched unrelated donor (n = 23) or a haploidentical family donor (n = 6). In four patients, an unmanipulated autologous graft was used. Infusion related toxicity due to the labeled antibody was minimal and no increase in treatment-related mortality due to the radioimmunoconjugate was observed. Day +30 and day +100 mortalities were 3 and 7%, respectively, and after a median follow up of 26 months treatment-related mortality was 30%. Late renal toxicity was observed in 14% of patients. The disease-free survival rate for 44 patients in 1 or 2 CR or in very good PR (< 15% blasts in the marrow at transplant) is 64% with only 8% disease-free survival for those with > 15% blasts in the marrow at transplant. PMID- 12533038 TI - Two-color flow cytometry with a CD19 gate for the evaluation of bone marrow involvement of B-cell lymphoma. AB - The usefulness of two-color flow cytometry with a CD19 gate (CD19-FCM) for detecting clonal B-cells in the bone marrow was evaluated. Since kappa and lambda ratios analyzed in the bone marrow from healthy adults were within the range of 0.5-3.0, outside of this range was regarded as the existence of clonal B-cells. Mixing experiments in vitro showed that 10% or more of clonal B-cells contaminating normal bone marrow cells could be detected using CD19-FCM. Ninety nine bone marrow samples from 60 patients with B-cell lymphoma were simultaneously analyzed using CD19-FCM and histologic examination of both bone marrow aspiration and core needle biopsy samples. The agreement of results obtained by both examinations was 82% (81/99). Discrepant results were found in 18 samples of 10 patients. All except one sample were positive for CD19-FCM analysis but negative for histologic examination and the conversion of CD19-FCM positivity to CD19-FCM-negativity was associated with chemotherapy. These results suggest that CD19-FCM is useful for the evaluation of bone marrow involvement of B-cell lymphoma. PMID- 12533039 TI - Carboxypeptidase-G2 rescue in cancer patients with delayed methotrexate elimination after high-dose methotrexate therapy. AB - High-dose methotrexate (HDMTX) is a component of many cancer treatment regimens. Despite careful management, delayed renal clearance, followed by extremely high serum levels with potentially life-threatening toxicity can occur. In the present study, we report our results of carboxypeptidase-G2 (CPDG2) rescue in 8 patients with delayed methotrexate elimination and renal impairment after HDMTX therapy for lymphoma or osteosarcoma. A dose of 50 U/kg CPDG2 was administered. MTX plasma levels decreased rapidly and recovery of renal function was observed in all patients. No patient developed severe WHO grade 4 MTX toxicity. CPDG2 provides an alternative route of MTX elimination by converting it to inactive and non-toxic metabolites. CPDG2 rescue was well tolerated, safe and very effective in preventing severe or life-threatening MTX toxicity. PMID- 12533040 TI - Primary myeloid sarcoma of the gynecologic tract: a report of two cases progressing to acute myeloid leukemia. AB - Primary gynecologic myeloid sarcomas are rare, and their diagnosis is often difficult. Differential diagnosis includes lymphomas and carcinomas of the gynecologic tract. We report the clinical, morphological, immunohistochemical and cytogenetic features of two cases of chloromas of the female genital tract, which progressed to acute myeloid leukemia in spite of aggressive therapy. PMID- 12533041 TI - Role of 18FDG-positron emission tomography scanning in the management of histiocytosis. AB - Diagnostic evaluation of histiocytic malignancies often involves a range of imaging studies to characterize skeletal and extraskeletal sites of involvement. Functional imaging with 18FDG PET provides a potential method for non-invasively detecting active disease. We report two cases where this modality was positive and facilitated therapeutic monitoring. PMID- 12533042 TI - Reverse seroconversion of hepatitis B virus after hematopoietic stem cell transplantation. AB - Hepatitis B virus (HBV) reactivation in patients previously positive for hepatitis B surface antibody (HBsAb), so-called reverse seroconversion, has been considered to be a rare complication after hematopoietic stem cell transplantation (HSCT). We experienced two patients who developed reverse seroconversion among nine who were HBsAb positive and Hepatitis B core antibody (HBcAb) positive before HSCT; one after autologous bone marrow transplantation (BMT) and another after allogeneic peripheral blood stem cell transplantation (PBSCT). We reviewed the literature and considered that reverse seroconversion of HBV after HSCT is not uncommon among HBsAb positive recipients. The use of corticosteroids, the lack of HBsAb in donor, and a decrease in serum HBsAb and HBcAb levels may predict reverse seroconversion after HSCT. PMID- 12533043 TI - SYK and LYN mediate B-cell receptor-independent calcium-induced apoptosis in DT 40 lymphoma B-cells. AB - Here, we report that the calcium ionophore ionomycin induces a massive Ca2+ dependent apoptosis in wildtype DT-40 chicken B lymphoma cells, as well as in BTK deficient, PLCgamma2-deficient and IP3 receptor-deficient DT-40 cells, but not in LYN- or SYK-deficient DT-40 cells. Notably, the deficiency of CSK, a negative regulator of Src-family PTK, promoted ionomycin-induced apoptosis of DT-40 cells. Reconstitution of SYK-deficient cells with wild-type SYK restored the apoptotic response of the cells to ionomycin, but the expression of FYN or LCK in LYN deficient cells did not restore the apoptotic response of LYN-deficient cells. Taken together, our data suggests that both LYN and SYK, but not BTK, FYN or LCK, are crucial mediators of BCR-independent Ca2+-induced apoptosis in DT-40 lymphoma B cells. PMID- 12533044 TI - Chemotherapy induces bcl-2 cleavage in lymphoid leukemic cell lines. AB - Bcl-2 is the major anti-apoptotic protein evaluated in studies aimed at understanding programmed cell death. Recent work suggests that the biological activity of Bcl-2 is modulated by proteolytic cleavage, with a 23 kDa cleaved Bcl 2 product having pro-apoptotic activity. In the current study we evaluated the effect of chemotherapy on Bcl-2 cleavage in B lineage leukemic cell lines. JM-1, SUP-B 15 and RS4 leukemic cell lines cleaved Bcl-2 to its 23 kDa form when exposed to the chemotherapeutic agents 1-beta-D-arabinofuranosyl-cytosine (Ara-C) or etoposide (VP-16). Chemotherapy induced Bcl-2 cleavage was blunted by inhibition of caspase activity. Co-culture of leukemic cells with bone marrow stromal cells during chemotherapy exposure resulted in reduced levels of 23 kDa Bcl-2 protein. These observations suggest that the bone marrow microenvironment may contribute to maintenance of residual leukemic disease during treatment by reducing generation of pro-apoptotic 23 kDa Bcl-2. PMID- 12533045 TI - A novel B-cell line (U-2932) established from a patient with diffuse large B-cell lymphoma following Hodgkin lymphoma. AB - Little is known about mechanisms leading to secondary non-Hodgkin lymphomas (NHL) in patients treated for Hodgkin lymphoma (HL). Our aim was to characterise in detail a cell line derived from a diffuse large B-cell lymphoma (DLBCL) that had developed in a patient with relapsing HL. The cell line U-2932 was established from ascites in a patient suffering from DLBCL previously treated for HL with multiple chemotherapy regimens. Characterisation was based on morphology, immunophenotype, Epstein-Barr virus (EBV)-status, IgH gene rearrangement status, tumourigenicity, p53 sequencing, and immunohistochemical expression of p53, BCL-2 and BCL-6. The karyotype was investigated using G-banding, comparative genomic hybridisation (CGH) and spectral karyotype (SKY) analysis. This cell line shows typical morphological features of a DLBCL and grows as colonies in nude mice. It expresses a B-cell phenotype with a somatically hypermutated V(H)4-39 gene and is negative for EBV. The origin of U-2932 was confirmed by demonstrating an identical V(H)4 rearrangement in ascites from the patient. A point mutation of the tumour-suppressor gene p53 was detected in amino acid position 176 and immunohistochemical over-expression of the p53 protein was also demonstrated. U 2932 carries a complex karyotype including high-level amplifications of the chromosomal bands 18q21 and 3q27 and expresses aberrant BCL-2 and BCL-6 immunohistochemically. We were unable to investigate the clonal relationship between the original HL and U-2932. In conclusion, U-2932 is a unique B cell line established from a patient suffering from HL followed by NHL. Overexpression of BCL-2, BCL-6 and p53 may play a role in the tumourigenesis and drug resistance. This cell line may become a useful tool to better understand the mechanisms responsible for development of secondary NHL in patients treated for HL. PMID- 12533046 TI - The cytotoxicity of arsenic trioxide to normal hematopoietic progenitors and leukemic cells is dependent on their cell-cycle status. AB - Arsenic trioxide (ATO) is a novel agent to treat acute promyelocytic leukemia (APL). ATO can degrade chimeric PML-RAR proteins and induce apoptosis in various cancer cells. However, its effects on primary hematopoietic CD34+ have not been examined. In this study, we compared the effects of ATO on HL60 leukemic cells and primary umbilical cord blood (UCB) CD34+ cells. HL60 cells and UCB CD34+ cells were cultured with different concentrations of ATO for up to three weeks and examined for changes of cell cycle. We found that ATO (< or = 5 microM) caused prolongation of G1/S and G2/M phase in a dose-dependent manner. The percentage of cells in G2/M increased significantly (from 8.6 to 53.8%). High dose ATO (> or = 25 microM) caused non-specific cell death in HL60 cells without any changes in cell cycle. In contrast to HL60 cells, UCB CD34+ cells were more resistant to high-dose ATO and most ATO-resistant CD34+ cells remained in G0/G1 phase. Primary cells that were resistant to ATO were rich in CD34+ cells. We further show that the ATO resistance was not related to the expression of P glycoprotein (MDR-1). Our results suggest that the resistance to ATO in primitive UCB CD34+ cells is most likely related to its cell-cycle status. These results could be useful to design treatments for non-APL malignancies and to enrich hematopoietic stem cells in clinically applicable settings. PMID- 12533047 TI - Mast cell heterogeneity in B-cell non-Hodgkin's lymphomas: an ultrastructural study. AB - Mast cells (MC) are critical for a number of pathological conditions, including acute and chronic inflammation and tumor angiogenesis. We have previously demonstrated that in B-cell non-Hodgkin's lymphoma (B-NHL) angiogenesis is correlated with total methachromatic and tryptase-positive MC and that both counts increase in step with the increase in malignancy, whereas the role of MC in malignant lymph nodes is not fully clear. An extensive ultrastructural study has been made of representative samples of 30 B-NHL and 10 benign lymphadenopathies. A heterogeneous population of MC characterized by the presence of granules with a semilunar aspect and containing scrolls was observed. The former are the expression of a slow but progressive release of angiogenic factors due to chronic, progressive stimulation of MC degranulation, while the latter contain tryptase, an angiogenic factor. These two ultrastructural data confirm the important role played by MC in the angiogenesis associated with progression in B-NHL. PMID- 12533048 TI - Hairy cell leukemia presenting as localized skeletal involvement. AB - We report a 45-year-old man who presented with localized skeletal involvement as the initial manifestation of hairy cell leukemia (HCL) without abnormal peripheral blood counts, splenomegaly or posterior iliac crest bone marrow involvement. The patient presented with pain in the left thigh. A plain radiograph was normal, but a magnetic resonance imaging (MRI) of this region showed a marrow-based lesion occupying the left femur neck, left proximal femur and both greater trochanters. Histological and immunophenotypic examination revealed a focal infiltrate of HCL. Skeletal involvement by HCL without co existing bone marrow involvement should be included in the differential diagnosis of bone marrow lesions where metastatic tumor is the foremost consideration. PMID- 12533049 TI - Osseous malignant non-Hodgkin's B-cell lymphoma associated with total hip replacement. AB - We report a case of non-Hodgkin B-cell lymphoma within the bony base of an implanted acetabular cup of a total hip arthroplasty. The diagnosis was made while revising a loose prosthesis and treatment of a non-union of a periprosthetic fracture. This case demonstrates that it is important to consider tumour development in differential diagnoses of pain originating in and around a loose prosthesis. It also illustrates the need for better understanding of the relationship between haematopoietic malignancies and possible carcinogenic effects of implant debris. PMID- 12533051 TI - Disseminated granulocytic sarcoma treated with allogeneic peripheral blood stem cell transplantation. AB - Primary granulocytic sarcoma is a rare disorder that presents as an extramedullary myeloid cell tumour. In this case, we describe a 35-year-old woman who developed widely disseminated granulocytic sarcomas three months after she received local radiotherapy to a primary granulocytic sarcoma of the cervical spine. These disseminated sarcomas initially responded to combination chemotherapy but this approach was only partly successful as the woman was left with increasing residual disease in the mediastinum. She was then treated with an allogeneic peripheral blood stem-cell transplant that led to complete resolution of her disease. This case demonstrates the effectiveness of allogeneic stem-cell transplantation in the treatment of progressive granulocytic sarcoma present after high-dose chemotherapy. PMID- 12533050 TI - Concomitant primary polycythemia vera and follicle center cell non-Hodgkin lymphoma: a case report and review of the literature. AB - The association of myeloproliferative and lymphoproliferative disorders is well known after cytotoxic drug or radiation exposure, while it is remarkably rare prior to therapy. We report on a patient simultaneously diagnosed as having polycythemia vera and II3A follicle center cell non-Hodgkin lymphoma (grade 1). At this timepoint, he is on 12-year follow-up, characterized by post-polycythemia myeloid metaplasia with myelofibrosis and persistent complete remission of lymphoma. The conventional marrow cytogenetic analysis performed during the course of the disease demonstrated an abnormal karyotype with deletion of the long arm of chromosome 20 and trisomy 8, while molecular analysis failed to detect BCR-ABL rearrangement in peripheral blood cells. To the best of our knowledge based on a computer-aided review of the literature (MED-LINE 1966 2002), this is the sixth case of concomitant primary polycythemia vera and lymphoma of non-Hodgkin type. Besides, there is a single literature report on polycythemia vera coexisting with the Hodgkin's lymphoma. In our case as well as in the recorded ones, two independent malignant clones of myeloid and lymphoid origin, respectively, seem to have arisen. Further reports, supported by chromosomal and molecular studies, could improve our knowledge on this extremely infrequent disease association. PMID- 12533052 TI - Chronic lymphocytic leukaemia developing in the course of chronic myeloid leukaemia. AB - Both chronic myeloid leukaemia (CML) and chronic lymphocytic leukaemia (CLL) are rare illnesses. Their co-existence has been previously reported, typically with CML following CLL. We report to our knowledge the first case of CLL developing some years after initial diagnosis of Philadelphia (Ph) chromosome positive CML. We provide molecular evidence that CLL arose in a Ph negative clone. The implications of the findings are discussed. PMID- 12533053 TI - Thymic epithelial hyperplasia with nodular sclerosis Hodgkin's disease. AB - We report a case of simultaneous occurrence of thymic epithelial hyperplasia and Hodgkin's disease. A computed tomographic scan of the chest revealed a tumor in the anterior mediastinum and conspicuous swelling of lymph nodes in the upper and lower mediastinum. The anterior mediastinal tumor was histologically diagnosed as thymic epithelial hyperplasia, while the lymph nodes were nodular sclerosis Hodgkin's disease. Our findings suggest that thymic hyperplasia might be directly involved in the pathology of at least some cases of Hodgkin's disease. PMID- 12533054 TI - More potent graft-versus-myeloma effect than graft-versus-renal cell cancer effect. PMID- 12533055 TI - Rituximab provides durable remission in a patient with refractory aggressive diffuse B-cell lymphoma failing salvage chemotherapy. AB - The outcome of patients with aggressive refractory diffuse large B-cell lymphoma (DLCL) is generally poor. A 43-year-old female with DLCL, who relapsed after first line chemotherapy (CHOP--cyclophosphamide, doxorubicin, vincristine, and prednisone) and progressed despite salvage chemotherapy (EPOCH-etoposide, prednisone, vincristine, cyclophosphamide, and doxorubicin), was treated effectively with 8 cycles of Rituximab. She is without evidence of disease with a follow-up of 32 months. We report this case to bring to attention the possibility of sustained durable remission with single agent Rituximab in refractory DLCL. PMID- 12533056 TI - HTLV-I carrier status disclosed at diagnosis of cytomegalovirus gastric ulcer. AB - A 50-year-old man experienced epigastric pain and gastrofiberscopy (GF) disclosed A2 to H1 stage multiple ulcerative lesions in the stomach. Inclusion bodies were found in the gastric mucosal cells of the biopsy specimen and the cells were positive for monoclonal antibody against cytomegalovirus. Serum HTLV-I antibody was positive and no atypical lymphocytes were seen in the peripheral blood. Gancyclovir and lansoprazole were given. A follow-up GF showed good ulcer healing with immunohistological improvement. The patient died from acute hepatic failure of undetermined cause eleven months after diagnosis. This is the first case report where a cytomegaloviral-induced gastric ulcer disclosed HTLV-I carrier state. PMID- 12533057 TI - Combination assay of urinary free L-fucose and trypsin inhibitor may be useful indicators of disease activity in patients with hematologic malignancies. AB - L-Fucose is a monosaccharide located at the non-reducing ends of sugar chains of glycoconjugates. Urinary L-fucose (U-FC) is excreted as free L-fucose, and clinically useful as a tumor marker of digestive organ cancers. We evaluated the clinical usefulness of U-FC levels in patients with various hematologic malignancies because U-FC for hematologic malignancies has only rarely been described. The mean U-FC levels in the acute nonlymphocytic leukemia (ANLL), myelodysplastic syndromes (MDS) and myeloproliferative disorders (MPD) groups were significantly higher than in the control group (P < 0.05). Recently, we reported that urinary trypsin inhibitor (UTI) levels in patients with ANLL, MDS, non-Hodgkin's lymphoma and multiple myeloma were significantly elevated, compared with those in healthy adult volunteers. Noninvasive combination assays of UTI and U-FC may have a higher accuracy in diagnosis of ANLL and MDS than those of UTI or U-FC alone. UTI and U-FC combination assays, noninvasive for patients, could be expanded as useful indicators in hematologic malignancies. PMID- 12533059 TI - Charity work--a professional's view. PMID- 12533058 TI - Impact of CD34 cell dose on acute graft versus host disease after allogeneic bone marrow stem cell transplantation. PMID- 12533060 TI - Trillium-coated oxygenators in adult open-heart surgery: a prospective randomized trial. AB - This randomized, prospective clinical trial examines the impact of the use of Trillium biopassive surface coating on clinical outcomes after cardiopulmonary bypass (CPB) that may be induced by contact of blood elements with foreign surfaces. The study consisted of 98 consecutive patients randomly assigned to either a CPB circuit that consisted of a Trillium-coated Affinity open reservoir oxygenator or a CPB circuit with an uncoated Affinity open reservoir oxygenator. The operative procedure performed on all 98 patients consisted of either coronary artery bypass graft (CABG), valve, or a combination of the two. Exclusion criteria consisted of patients who presented to the operating room in circulatory arrest. Trillium biopassive surface coating resulted in improved clinical outcomes and fewer adverse events when compared to the control group. Significantly, fewer patients required no blood products (18.3% in the control group vs. 32.7% in the treatment group), even though the control group had a significantly higher pre-bypass hematocrit. Postoperative atrial fibrillation (24.5% vs. 16.3%) and reoperation for bleeding (10.2% vs. 4.1%) showed a much lower incidence in the Trillium group. Significance was not reached because of the small sample size resulting in low power. Trillium circuits result in improved patient outcomes in the treatment group when compared to the control circuit group. PMID- 12533061 TI - High-volume, zero balanced ultrafiltration improves pulmonary function in a model of post-pump syndrome. AB - The systemic inflammatory response syndrome (SIRS), which may develop following cardiopulmonary bypass (CPB), can cause postoperative complications that contribute to the morbidity and mortality associated with open-heart surgery. Inflammatory mediators such as cytokines, are thought to play an important role in SIRS. Zero Balance Ultrafiltration (Z-BUF) is thought to reduce the quantity of inflammatory mediators associated with CPB and may attenuate the adverse effects of bypass. Following ethics committee approval, both an unfiltered experimental group and Z-BUF treatment group consisting of Yorkshire pigs (41 +/- 19 kg) were anesthetized, ventilated, instrumented, cannulated and placed on CPB for 60 minutes. Following CPB, an infusion of low-dose endotoxin (1 microg/kg) was administered I.V. and the animals were monitored for 3.5 hours. The Z-BUF treatment group (n = 5) received high-volume Z-BUF (122 ml/kg +/- 41) and the unfiltered experimental group (n = 5) did not. Hemodynamics, blood gases, and pulmonary functions were measured before, during, and after CPB. Following euthanasia, the middle lobe of the lung was prepared for histological analysis. Necropsy of the lung sample was weighed before and after dehydration to evaluate lung water content. During the experimental time course, plasma samples were evaluated for Interleukin-8 (IL-8) concentrations. Arterial PO2's (mmHg) in the unfiltered experimental group showed a significant reduction at 3.5 hours post CPB when compared to baseline while the Z-BUF treatment group PaO2 did not significantly change. There was a significant difference in the PaO2 between the unfiltered experimental and Z-BUF group at the final 3.5 hour time point (78 +/- 32 vs. 188 +/- 92 mmHg respectively). Pulmonary compliance (ml/cmH2O) was significantly reduced in both the unfiltered experimental and Z-BUF treatment groups with the unfiltered experimental group being the most significant. Lung wet/dry ratios were established and results found the unfiltered experimental group ratio significantly greater than that of the Z-BUF treatment group. Morphometric analysis of histologic lung sections confirmed pulmonary injury in the unfiltered experimental group and protection in the Z-BUF treatment group. This study suggests that Z-BUF provides higher arterial PO2's and lung compliances while reducing pulmonary edema and lung injury in a porcine model of PPS. PMID- 12533062 TI - Coronary artery bypass grafting: an off-pump versus on-pump review. AB - There has been a proliferation in the number of coronary artery bypass grafts (CABG) being performed without the use of cardiopulmonary bypass (CPB). However, the benefits of off-pump coronary artery grafting (OPCAB) are still being determined. The aim of this retrospective review was to compare the perioperative outcomes of CPB patients with OPCAB patients and to identify the patients most likely to benefit from the off-pump procedure. We reviewed the perioperative data of all isolated CABG patients at two metropolitan hospitals for the period of August 2000 to September 2001. The two groups (OPCAB vs. CPB) were further divided into subgroups identifying patients by their predicted mortality (higher risk and lower-risk) and the number of distal graft anastomoses received (1, 2, 3, 4, or 5). A p value less than .05 was considered significant. Out of the total of 882 patients, 46.2% were OPCAB cases. Both CPB and OPCAB groups were similar in terms of demographics and predicted risk of mortality. Intraoperatively, OPCAB patients had fewer distal graft anastomoses (2.4 +/- 1.0 vs. 3.2 +/- 1.0, p < .001). Postoperatively, patients in the OPCAB group had less chest drainage (889 +/- 588 vs. 989 +/- 662 mls, p < .001), sustained fewer strokes (0.2 vs. 1.9%, p < .05), were transfused less (15.4 vs. 32.5%, p < .001) and were discharged earlier (7.3 +/- 5.6 vs. 8.5 +/- 9.1 days, p < .05). For higher-risk patients, OPCAB was associated with fewer reoperations for bleeding (1.3 vs. 6.4%, p < .05), a lower stroke rate (0 vs. 3.2%, p < .05), and a trend toward lower mortality (7.1 vs. 15.1%, p = .08). However, lower-risk OPCAB patients' stroke incidences (0.5% OPCAB group vs. 1.4% CPB group), and mortality rates (0.5 vs. 0.5%) were similar. Comparisons by number of grafts performed revealed that only the single-grafted OPCAB patients had statistically fewer postoperative complications, reduced chest drainage, and a shorter intensive care stay. Differences between either operation groups in transfusion rates were only statistically significant for the one to three grafted patients, while postoperative stays were similar for patients having four grafts. These results suggest that OPCAB is associated with a reduction in mortality and morbidity, particularly within the higher-risk patients. However, the benefits of OPCAB diminished with an increasing number of distal anastomoses performed. PMID- 12533063 TI - The effect of surface modification and aprotinin on cellular injury during simulated cardiopulmonary bypass. AB - Cardiopulmonary bypass (CPB) elicits derangements to the formed elements of blood because of the physical stresses of extracorporeal flow. Methods of reducing the impact of CPB include circuit surface modification and pharmacological supplementation. The purpose of this study was to examine the effects of aprotinin in combination with surface modification during simulated CPB. Fresh whole bovine blood was used to prime standard CPB circuits divided into four groups (N = 3): control (CTR), aprotinin 300 KIU/mL (APR), Poly (2 methoxyethylacrylate) coating (PMEA), and APR with PMEA (APR-PMEA). Physical stresses included venous reservoir negative pressure (-85 mmHg), arterial line pressure of 150 mmHg at 5 LPM, and air-blood interface, applied over a 90-minute period. Samples were drawn at the following times: 0, 10, 45, and 90 minutes. Endpoints included platelet count (PLT), plasma-free hemoglobin (PFHb), and thromboelastography (TEG). PLT did not change (138.9 +/- 15.0 vs. 102.9 +/- 21.0, p = ns) throughout the 90-minute experimental periods in any group. PFHb increased significantly (mean of 19- fold) throughout the experiment, but was not affected by any treatment. The TEG index declined in the CTR (3.6 +/- 0.4 vs. 16.2 +/- 2.9, p < .0003), PMEA (5.9 +/- 0.8 vs. -2.7 +/- 3.8, p < .02), and APR PMEA (4.6 +/- 1.0 vs. -2.8 +/- 0.3 p < .0003) groups, but not in the APR group (3.6 +/- 2.2 vs. -1.3 +/- 3.3 p = .10). In conclusion, neither APR nor PMEA had an effect on either red cell hemolysis or PLT, but APR treatment alone significantly attenuated the derangements in coagulation induced in this extracorporeal model. PMID- 12533064 TI - Validation of a new whole blood coagulation monitoring system. AB - The Hemochron Response is a third generation point-of-care (POC) whole blood coagulation analyzer that retains the clinical utility of the Hemochron standard (801/8000) while providing a data management program that assists the POC coordinator with Quality Assurance (QA) compliance. Clinical and laboratory studies were performed to ensure consistency of the target anticoagulation times with the Hemochron standard and to evaluate precision and reproducibility of the Hemochron Response. Clinical tests for prothrombin time (PT) using fresh and citrated whole blood, activated clotting time (ACT), and activated partial thromboplastin time (APTT) showed excellent correlation to the Hemochron standard where r = 0.929, r = 0.969, r = 0.947, and r = 0.992, respectively. This was confirmed by a paired Student's t-test. The standard expectation for reproducibility of ACT tests has been a coefficient of variation (CV) of 10%. Laboratory studies of reproducibility and precision for the Response instrument included analysis of the CV using ACT test tubes. For normal and abnormal control plasma (CPL), the range of CVs observed was 3.3%-4.6% and 3.0%-5.0%, respectively. For heparin dose response analysis, the range for Donor 1 and 2 was 1.0%-4.2% and 1.1%-8.0%, respectively. These data suggest that the Hemochron Response is reliable and equivalent to the Hemochron standard in clinical applications. PMID- 12533065 TI - The heater-cooler unit--a conceivable source of infection. AB - Even drinking water is contaminated with pathogenic microorganisms. This does not necessarily pose a risk for healthy individuals, but it may result in serious consequences in people with impaired immune systems. This is particularly valid if drinking water is used for medical purposes. The heater-cooler unit (HCU) connected to heat exchangers or blankets by tubing, the connection is closed water circuit that contains microorganisms and algae. While connecting the tubing to the heat exchanger, spilling of water cannot be avoided. Microbiological examinations showed that germs and particles pollute the units. Exposure to the patient and the OR equipment has the potential to increase the risk of infection should the HCU water come in contact with the patient. As a result of the high incidence of particle and algae in the HCU, malfunction occurs. Sampling shows >1000/mL CFU (colony forming units) at 36 degrees C and 55/mL CFU at 20 degrees C on average. The specific findings include Pseudomonas and Legionella. Disinfecting HCU is very difficult. Often HCUs do not provide any technology to reduce bacterial or other contamination. The instructions for use of oxygenators often exclude the use of disinfectants. Maintenance instructions for the HCU advocate the use of disinfectants that carry the risk of oxygenator damage and of heat exchanger leakage. The effect of chemical disinfectants and heat exchanger membranes have not been examined, they may impair heat exchanger permeability and function. As an alternative to chemical and thermal disinfection, we used the alternative method of filtration. Using a membrane filter element, we noticed a decreasing number of CFUs from 55 to sterile conditions at 20 degrees C and from >1000 CFUs to 100 CFUs at 36 degrees C (Figure 1). In addition, we noticed a removal of other particles and algae. In conclusion, we have demonstrated a technique that is simple to implement and effectively reduces the microbiological load of the water in the heater-cooler unit. PMID- 12533066 TI - Argatroban in adult extracorporeal membrane oxygenation. AB - This case report addresses the use of Argatroban, an anticoagulant and thrombin inhibitor for treatment of thrombocytopenia in an adult patient on extracorporeal membrane oxygenation (ECMO). After 5 days on ECMO, the patient showed signs of heparin-induced thrombocytopenia (HIT) with a platelet count of 20K. Argatroban was initiated to decrease progression of HIT and continue treatment with ECMO proved to be successful. Given the occurrence of HIT with heparin therapy, a need for alternate drug therapy is required for patients requiring treatment with ECMO. The use of Argatroban in adult ECMO is outlined and includes dosage, monitoring, and patient treatment. PMID- 12533067 TI - A reusable training circuit for cardiopulmonary bypass. AB - At the turn of the millennium, perfusion teaching programs are faced with significant difficulties. The number of students in pediatric perfusion training has increased, and more importantly, the number of pediatric open-heart procedures has decreased because of a variety of reasons. Hence, they could barely satisfy the minimum requirements of pediatric cases established by the teaching programs. The idea of "a teaching circuit" that could reproduce and simulate cardiopulmonary bypass was designed. The trainee is able to manipulate the cardiopulmonary bypass circuit according to patient responses, to perform perfusion related-maneuvers in establishing and maintaining hemodynamic stability. The aim of this study was to simulate a patient on CPB, maintain stability during varying clinical situations, and then to wean the patient off CPB. The equipment used was reusable and nonsterile. PMID- 12533068 TI - The false promise of sustainable pumping rates. PMID- 12533069 TI - New horizons for the corporate hydrologist. PMID- 12533070 TI - A finite world, earth sciences, and public trust. AB - The beginning of the 21st century has coincided with our recognition that life sustaining earth cycles are remarkably fine-tuned, and that humans have developed technological abilities to perturb these cycles. Also, inspired bythe gifts of freedom and democracy, humans have given themselves laws to exploit nature for profit. The upshot is that nature's balance, governed by immutable physical laws, is being confronted by social laws driven by human aspirations. This conflict and its implications to the human relevance of the earth sciences are explored in the context of an extraordinary tradition of European culture known as public trust. PMID- 12533071 TI - A comparison of two stochastic inverse methods in a field-scale application. AB - Inverse modeling is a useful tool in ground water flow modeling studies. The most frequent difficulties encountered when using this technique are the lack of conditioning information (e.g., heads and transmissivities), the uncertainty in available data, and the nonuniqueness of the solution. These problems can be addressed and quantified through a stochastic Monte Carlo approach. The aim of this work was to compare the applicability of two stochastic inverse modeling approaches in a field-scale application. The multi-scaling (MS) approach uses a downscaling parameterization procedure that is not based on geostatistics. The pilot point (PP) approach uses geostatistical random fields as initial transmissivity values and an experimental variogram to condition the calibration. The studied area (375 km2) is part of a regional aquifer, northwest of Montreal in the St. Lawrence lowlands (southern Quebec). It is located in limestone, dolomite, and sandstone formations, and is mostly a fractured porous medium. The MS approach generated small errors on heads, but the calibrated transmissivity fields did not reproduce the variogram of observed transmissivities. The PP approach generated larger errors on heads but better reproduced the spatial structure of observed transmissivities. The PP approach was also less sensitive to uncertainty in head measurements. If reliable heads are available but no transmissivities are measured, the MS approach provides useful results. If reliable transmissivities with a well inferred spatial structure are available, then the PP approach is a better alternative. This approach however must be used with caution if measured transmissivities are not reliable. PMID- 12533072 TI - Radar determination of the spatial structure of hydraulic conductivity. AB - Spatial variability of hydraulic conductivity exerts a predominant control on the flow of fluid through porous media. Heterogeneities influence advective pathways, hydrodynamic dispersion, and density-dependent dispersion; they are, therefore, a key concern for studies of ground water resource development, contaminant transport, and reservoir engineering. Ground-penetrating radar contributes to the remote, geophysical characterization of the macroscale variability of natural porous media. On a controlled excavation of a glacial-fluvial sand and gravel deposit in the Fanshawe Delta area (Ontario, Canada), the hydraulic conductivity field of a 45 x 3 m vertical exposure was characterized using constant-head permeameter measurements performed on undisturbed horizontal sediment cores. Ground-penetrating radar data were collected along the excavation face in the form of both reflection and common midpoint surveys. Comparison of geostatistical analyses of the permeameter measurements and the radar data suggests thatthe horizontal correlation structure of radar stack velocity can be used to directly infer the horizontal correlation structure of hydraulic conductivity. The averaging nature of the common midpoint survey is manifest in the vertical correlation structure of stack velocity, making it less useful. Radar reflection data do not exhibit a spatial structure similar to that of hydraulic conductivity possibly because reflections are a result of material property contrasts rather than the material properties themselves. PMID- 12533073 TI - Deriving TDS values in coarse sediments from long normal and electromagnetic logs. AB - This paper presents an empirical relationship of quantitatively linked electromagnetic (EM) borehole recordings of the total dissolved solids (TDS) in pore water in the Quaternary deposits of the Belgian coastal plain. First, the long normal (LN) logs are linked to EM logs, then the already developed relationships between LN resistivity measurements and the TDS values are rewritten for EM recordings. The main parameter in these equations is the formation factor, which is derived from ground water analyses and LN logs through Archie's law. The EM recording has several advantages compared to the LN logs. The EM analysis allows measuring in PVC-cased wells and is not hindered by the invasion zone around the well. Furthermore, it has a high vertical resolution. LN logs can be measured only once, after drilling a well; EM recordings can be repeated several times in monitoring wells, which allows the gathering of time dependent data over a complete vertical cross section. Such data could be obtained with LN logs only in wells with screens over the full-depth interval, which causes a hydraulic short circuit. This short circuit can result in a large artificial flow through the well between different levels, resulting in a salinity profile, which is no longer representative for the studied site. Remediation against short circuiting is a reduction of the screened interval, which strongly reduces the gathered information. The application of the derived equations is one of setting up a monitoring network along the Belgian coast to monitor the trend in salinity levels and comparing present salinity levels with older LN recordings to investigate the salinity changes in the last 30 years. Deep wells already present in the Belgiancoastal plain can then be used to monitor both the fresh water head changes and the salt water evolution. The technique has also been used for parameter identification for which real concentration measurements were needed. PMID- 12533074 TI - Influence of aquifer properties on phytoremediation effectiveness. AB - Recent research has shown that planting deep-rooted trees, such as poplar, can take up and degrade important ground water pollutants such as trichloroethylene (TCE) as they transpire water from the capillary fringe of shallow contaminated aquifers. The effect of hydrogeologic factors on the minimum plantation area needed to prevent downgradient migration of contaminated ground water is not well known. Accordingly, the objective of this research was to identify the hydrogeologic parameters that control phytoremediation effectiveness. We used a numerical ground water flow model to evaluate the effect that natural variations in hydrogeologic parameters and growing season duration have on the minimum plantation area required for capture. We found that the plantation area that was needed to completely capture a ground water contamination plume was directly proportional to aquifer horizontal hydraulic conductivity, saturated thickness, and ground water gradient. The plantation area needed for capture increased nonlinearly with increasing plume width, aquifer anisotropy, and decreasing growing season duration. The plantation area needed for capture was generally insensitive to aquifer-specific yield and storativity. Steady-state simulations can be used to predict the plantation area needed for capture in many applications. A particularly important finding of this work is that evapotranspiration fluxes through plantations appropriately sized to contain the plume substantially exceeded the ground water flux through the plume itself. PMID- 12533075 TI - Enhanced attenuation of septic system phosphate in noncalcareous sediments. AB - Review of phosphate behavior in four mature septic system plumes on similar textured sand has revealed a strong correlation between carbonate mineral content and phosphate concentrations. A plume on calcareous sand (Cambridge site, 27 wt % CaCO3 equiv.) has proximal zone PO4 concentrations (4.8 mg/L P average) that are about 75% of the septic tank effluent value, whereas three plumes on noncalcareous sand (Muskoka, L. Joseph, and Nobel sites, <1 wt % CaCO3 equiv.) have proximal zone phosphate concentrations (<0.1 mg/L P) that are consistently less than 2% of the effluent values. Phosphate attenuation at the noncalcareous sites appears to be an indirect result of the development of acidic conditions (site average pH 3.5 to 5.9) and elevated Al concentrations (up to 24 mg/L), which subsequently causes the precipitation of Al-P minerals such as variscite (AlPO4 x 2H2O). This is supported by scanning electron microscope analyses, which show the widespread occurrence of (Al+P)--rich secondary mineral coatings on sand grains below the infiltration beds. All of these septic systems are more than 10 years old, indicating that these attenuation reactions have substantial longevity. A field lysimeter experiment demonstrated that this reaction sequence can be readily incorporated into engineered waste water treatment systems. We feel this important P removal mechanism has not been adequately recognized, particularly for its potential significance in reducing P loading from septic systems in lakeshore environments. PMID- 12533076 TI - Hydrostratigraphic modeling of a complex, glacial-drift aquifer system for importation into MODFLOW. AB - Deposition from at least three episodes of glaciation left a complex glacial drift aquifer system in central Illinois. The deepest and largest of these aquifers, the Sankoty-Mahomet Aquifer, occupies the lower part of a buried bedrock valley and supplies water to communities throughout central Illinois. Thin, discontinuous aquifers are present within glacial drift overlying the Sankoty-Mahomet Aquifer. This study was commissioned by local governments to identify possible areas where a regional water supply could be obtained from the aquifer with minimal adverse impacts on existing users. Geologic information from more than 2,200 existing water well logs was supplemented with new data from 28 test borings, water level measurements in 430 wells, and 35 km of surface geophysical profiles. A three-dimensional (3-D) hydrostratigraphic model was developed using a contouring software package, a geographic information system (GIS), and the 3-D geologic modeling package, EarthVision. The hydrostratigraphy of the glacial-drift sequence was depicted as seven uneven and discontinuous layers, which could be viewed from an infinite number of horizontal and vertical slices and as solid models of any layer. Several iterations were required before the 3-D model presented a reasonable depiction of the aquifer system. Layers from the resultant hydrostratigraphic model were imported into MODFLOW, where they were modified into continuous layers. This approach of developing a 3-D hydrostratigraphic model can be applied to other areas where complex aquifer systems are to be modeled and is also useful in helping lay audiences visualize aquifer systems. PMID- 12533077 TI - A statistical estimator of the spatial distribution of the water-table altitude. AB - An algorithm was designed to statistically estimate the areal distribution of water-table altitude. The altitude of the water table was bounded below by the minimum water-table surface and above by the land surface. Using lake elevations and stream stages, and interpolating between lakes and streams, the minimum water table surface was generated. A multiple linear regression among the minimum water table altitude, the differerence between land-surface and minimum water-table altitudes, and the water-level measurements from surficial aquifier system wells resulted in a consistently high correlation for all groups of physiographic regions in Florida. A simple linear regression between land-surface and water level measurements resulted in a root-mean-square residual of 4.23 m, with residuals ranging from -8.78 to 41.54 m. A simple linear regression between the minimum water table and the water-level measurements resulted in a root-mean square residual of 1.45 m, with residuals ranging from -7.39 to 4.10 m. The application of the multiple linear regression presented herein resulted in a root mean-square residual of 1.05 m, with residuals ranging from -5.24 to 5.63 m. Results from complete and partial F tests rejected the hypothesis of eliminating any of the regressors in the multiple linear regression presented in this study. PMID- 12533078 TI - Variability and comparison of hyporheic water temperatures and seepage fluxes in a small Atlantic salmon stream. AB - Ground water discharge is often a significant factor in the quality of fish spawning and rearing habitat and for highly biologically productive streams. In the present study, water temperatures (stream and hyporheic) and seepage fluxes were used to characterize shallow ground water discharge and recharge within thestreambed of Catamaran Brook, a small Atlantic salmon (Salmo salar) stream in central New Brunswick, Canada. Three study sites were instrumented using a total of 10 temperature sensors and 18 seepage meters. Highly variable mean seepage fluxes, ranging from 1.7 x 10(-4) to 2.5 cm3 m(-2) sec(-1), and mean hyporheic water temperatures, ranging from 10.5 degrees to 18.0 degrees C, at depths of 20 to 30 cm in the streambed were dependent on streambed location (left versus right stream bank and site location) and time during the summer sampling season. Temperature data were usefulfor determining if an area of the streambed was under discharge (positive flux), recharge (negative flux), or parallel flow (no flux) conditions and seepage meters were used to directly measure the quantity of water flux. Hyporheic water temperature measurements and specific conductance measurements of the seepage meter sample water, mean values ranging from 68.8 to 157.9 microS/cm, provided additional data for determining flux sources. Three stream banks were consistently under discharge conditions, while the other three stream banks showed reversal from discharge to recharge conditions over the sampling season. Results indicate that the majority of the water collected in the seepage meters was composed of surface water. The data obtained suggests that even though a positive seepage flux is often interpreted as ground water discharge, this discharging water may be of stream water origin that has recently entered the hyporheic zone.The measurement of seepage flux in conjunction with hyporheic water temperature or other indicators of water origin should be considered when attempting to quantify the magnitude of exchange and the source of hyporheic water. PMID- 12533079 TI - Salinization of a fresh palaeo-ground water resource by enhanced recharge. AB - Deterioration of fresh ground water resources caused by salinization is a growing issue in many arid and semi-arid parts of the world. We discuss here the incipient salinization of a 10(4) km2 area of fresh ground water (<3,000 mg/L) in the semiarid Murray Basin of Australia caused by widespread changes in land use. Ground water 14C concentrations and unsaturated zone Cl soil water inventories indicate that the low salinity ground water originated mainly from palaeo recharge during wet climatic periods more than 20,000 years ago. However, much of the soil water in the 20 to 60 m thick unsaturated zone throughout the area is generally saline (>15,000 mg/L) because of relatively high evapotranspiration during the predominantly semiarid climate of the last 20,000 years. Widespread clearing of native vegetation over the last 100 years and replacement with crops and pastures leads to enhancement of recharge rates that progressively displace the saline soil-water from the unsaturated zone into the ground water. To quantify the impact of this new hydrologic regime, a one-dimensional model that simulates projected ground water salinities as a function of depth to ground water, recharge rates, and soil water salt inventory was developed. Results from the model suggest that, in some areas, the ground water salinity within the top 10 m of the water table is likely to increase by a factor of 2 to 6 during the next 100 years. Ground water quality will therefore potentially degrade beyond the point of usefulness well before extraction of the ground water exhausts the resource. PMID- 12533080 TI - Aquifer parameter estimation from surface resistivity data. AB - This paper is devoted to the additional use, other than ground water exploration, of surface geoelectrical sounding data for aquifer hydraulic parameter estimation. In a mesoscopic framework, approximated analytical equations are developed separately for saline and for fresh water saturations. A few existing useful aquifer models, both for clean and shaley sandstones, are discussed in terms of their electrical and hydraulic effects, along with the linkage between the two. These equations are derived for insight and physical understanding of the phenomenon. In a macroscopic scale, a general aquifer model is proposed and analytical relations are derived for meaningful estimation, with a higher level of confidence, of hydraulic parameter from electrical parameters. The physical reasons for two different equations at the macroscopic level are explicitly explained to avoid confusion. Numerical examples from existing literature are reproduced to buttress our viewpoint. PMID- 12533081 TI - The journal of 'ground water': in the beginning. PMID- 12533083 TI - Cyp46 (24S-cholesterol hydroxylase): a genetic risk factor for Alzheimer disease. PMID- 12533084 TI - Neurological aspects of biological and chemical terrorism: a review for neurologists. AB - The Centers for Disease Control and Prevention urge physicians to become familiar with chemical and biological weapons. Preparedness among neurologists is especially important because several of these agents affect the nervous system. This article reviews 4 agents that have a history of military or terrorist use: cyanide poisons, organophosphate poisons, botulinum toxin, and anthrax. Cyanide and organophosphate poisons are characterized by dose-dependent impairment of neurological function with nonspecific symptoms such as headache or dizziness at one end of the spectrum and convulsions and coma at the other. Neurological examinations help clinicians to differentiate these agents from other intoxications. Botulinum toxin has a delayed onset of action and results in descending paralysis and prominent cranial nerve palsies. Anthrax frequently causes fulminating hemorrhagic meningitis. Early recognition of these chemical and biological weapons is key to instituting specific therapy and preventing casualties within the health care team and the community at large. PMID- 12533085 TI - Increased brain beta-amyloid load, phosphorylated tau, and risk of Alzheimer disease associated with an intronic CYP46 polymorphism. AB - BACKGROUND: CYP46, the gene encoding cholesterol 24-hydroxylase, plays a key role in the hydroxylation of cholesterol and thereby mediates its removal from brain. OBJECTIVE: To study the association of polymorphic sites on CYP46 with Alzheimer disease (AD) traits and with the risk of the development of AD. DESIGN: Alzheimer disease traits (beta-amyloid load, beta-amyloid peptides, hyperphosphorylated tau protein) were assessed in brain tissues and in the cerebrospinal fluid of patients with AD and control subjects. Genetic associations were studied in 2 independent populations. SETTING: Specialized centers for memory disorders in Switzerland, Greece, and Italy. PARTICIPANTS: Fifty-five brain tissues from nondemented elderly patients for the histopathological studies; 38 patients with AD and 25 control subjects for the cerebrospinal fluid studies; 201 patients with AD and 248 control subjects for the genetic association studies. RESULTS: A polymorphism of CYP46 was associated with increased beta-amyloid load in brain tissues as well as with increased cerebrospinal fluid levels of beta-amyloid peptides and phosphorylated tau protein. Moreover, this CYP46 polymorphism was associated with higher risk of late-onset sporadic AD in 2 independent populations (odds ratio, 2.16; 95% confidence interval [CI], 1.41-3.32; P<.001). The additional presence of 1 or 2 apolipoprotein E epsilon4 alleles synergistically increased the risk of AD to an odds ratio of 9.6 (95% CI, 4.9 18.9; P<.001) as compared with 4.4 for apolipoprotein E epsilon4 alone (95% CI, 2.8-6.8; P<.001). CONCLUSION: CYP46 influences brain beta-amyloid load, cerebrospinal fluid levels of beta-amyloid peptides and phosphorylated tau, and the genetic risk of late-onset sporadic AD. PMID- 12533086 TI - Prediction of early clinical severity and extent of neuronal damage in anterior circulation infarction using the initial serum neuron-specific enolase level. AB - CONTEXT: Prompt and precise measurement of neuronal damage in acute cerebral infarction is important to determine the prognosis of functional outcome. A feasible biochemical marker such as the neuron-specific enolase (NSE) level has been used to detect various diseases involving the central nervous system. OBJECTIVE: To determine whether the initial serum NSE level is a useful marker for predicting the severity of clinical neurological deficits and the extent of neuronal damage in acute anterior-circulation infarction. DESIGN: Case-control study with biochemical-clinicoradiological correlation. SETTING: Tertiary care center. PARTICIPANTS: Eighty-one patients and 77 age- and sex-matched control subjects. MAIN OUTCOME MEASURES: Patients with anterior-circulation infarction underwent intravenous serum NSE sampling within 24 hours after symptom onset. Recent infarction was confirmed by T2-weighted and diffusion-weighted magnetic resonance imaging of the brain about 1 week after the onset of stroke. Volumetric analysis of infarction was also performed. The National Institutes of Health Stroke Scale score was measured on admission to the hospital and 1 week after symptom onset. RESULTS: The patients' initial serum NSE levels were statistically significantly higher than the controls (P<.05). The initial serum NSE level highly correlated with the volume of infarction seen on T2-weighted magnetic resonance imaging of the brain (r = 0.62, P<.001) and with the National Institutes of Health Stroke Scale score obtained on hospital admission (r = 0.42, P =.002) and on the seventh day after the onset of stroke (r = 0.44, P<.001). CONCLUSION: The initial serum NSE level is a reliable predictor for the extent of neuronal damage and the severity of clinical neurological deficits in acute anterior-circulation infarction. PMID- 12533087 TI - Antiganglioside antibodies in multifocal acquired sensory and motor neuropathy. AB - BACKGROUND: Multifocal acquired demyelinating sensory and motor neuropathies are considered autoimmune and responsive to immunotherapy. In the absence of demyelination, however, they are considered idiopathic if no other cause is found. OBJECTIVE: To determine whether patients with multifocal acquired sensory and motor neuropathy of an otherwise unknown cause have antiganglioside antibodies, regardless of whether they are classified as demyelinating or axonal, indicating a possible immune-mediated origin. PATIENTS AND METHODS: Serum samples from 25 patients with multifocal acquired sensory and motor neuropathy of an otherwise unknown cause were tested for antibodies to gangliosides using an agglutination immunoassay. Reactive serum samples were further tested by enzyme linked immunosorbent assay against individual gangliosides. Electrophysiologic studies were reviewed for evidence of demyelination. RESULTS: Increased levels of ganglioside antibodies were detected in 12 (48%) of the 25 patients using the agglutination immunoassay, and in 7 (58%) of the 12 agglutination-positive patients by the enzyme-linked immunosorbent assay. Serum samples from these 7 patients had IgG antibodies to 1 or more gangliosides; none had elevated levels of IgM antiganglioside antibodies. Three of the patients fulfilled 2 of the American Academy of Neurology electrophysiologic criteria for demyelination, but none fulfilled the 3 of the 4 possible criteria required for the diagnosis of demyelinating neuropathy. A sural nerve biopsy specimen in 2 patients revealed axonal degeneration. CONCLUSION: Multifocal sensory and motor neuropathies of an otherwise unknown cause may be associated with antiganglioside antibodies, regardless of whether they exhibit demyelinating features. PMID- 12533088 TI - Increased metabolic muscle fatigue is caused by some but not all mitochondrial mutations. AB - CONTEXT: Excessive muscle fatigue occurs in patients with a mitochondrial encephalomyopathy (MEM), but it is also a frequent problem in patients with other neuromuscular disorders (ONMD). OBJECTIVE: To determine whether, and to what extent, metabolic muscle fatigue specifically occurs in patients with an MEM. DESIGN: Metabolic muscle fatigue was assessed in a series of 21 patients with an MEM, including 13 patients with chronic progressive external ophthalmoplegia and 8 patients with various mitochondrial point mutations; 27 patients with ONMD; and 25 healthy controls. Isometric twitch force of the ankle dorsiflexors was measured after supramaximal stimulation of peroneal nerves. Six trains of stimuli (of 1 minute's duration with rates from 0.2 to 5 stimuli per second) were given to each subject. RESULTS: An abnormal decrement of the twitch amplitude that occurred during a stimulation train was found in patients with MEM and in those with ONMD. The decrement of the twitch amplitude of controls and of patients with ONMD was strongly influenced by their muscle force (P<.001). After subtraction of the influence of the muscle force, specific fatigue was notably higher in patients with chronic progressive external ophthalmoplegia than in patients with ONMD and in controls, and it correlated well with elevations of serum lactate. Specific fatigue was also abnormal in a patient with a mitochondrial G7497A mutation, but normal in patients with an A3243G or a G11778A mutation. The heteroplasmy of mitochondrial DNA in muscle correlated neither with the force measures nor with the serum lactate levels. CONCLUSIONS: Generally, metabolic muscle fatigue accompanies muscular weakness. Specifically, some but not all mitochondrial mutations cause excessive metabolic muscle fatigue. PMID- 12533089 TI - Elevated plasma homocysteine levels in patients treated with levodopa: association with vascular disease. AB - BACKGROUND: Hyperhomocysteinemia is a risk factor for vascular disease and potentially for dementia and depression. The most common cause of elevated homocysteine levels is deficiency of folate or vitamin B(12). However, patients with Parkinson disease (PD) may have elevated homocysteine levels resulting from methylation of levodopa and dopamine by catechol O-methyltransferase, an enzyme that uses S-adenosylmethionine as a methyl donor and yields S adenosylhomocysteine. Since S-adenosylhomocysteine is rapidly converted to homocysteine, levodopa therapy may put patients at increased risk for vascular disease by raising homocysteine levels. OBJECTIVES: To determine whether elevations in plasma homocysteine levels caused by levodopa use are associated with increased prevalence of coronary artery disease (CAD), and to determine what role folate and vitamin B(12) have in levodopa-induced hyperhomocysteinemia. DESIGN/METHODS: Subjects included 235 patients with PD followed up in a movement disorders clinic. Of these, 201 had been treated with levodopa, and 34 had not. Blood samples were collected for the measurement of homocysteine, folate, cobalamin, and methylmalonic acid levels. A history of CAD (prior myocardial infarctions, coronary artery bypass grafting, or coronary angioplasty procedures) was prospectively elicited. We analyzed parametric data by means of 1-way analysis of variance or the t test, and categorical data by means of the Fisher exact test or chi(2) test. RESULTS: Mean +/- SD plasma homocysteine levels were significantly higher in patients treated with levodopa (16.1 +/- 6.2 micro mol/L), compared with levodopa-naive patients (12.2 +/- 4.2 micro mol/L; P<.001). We found no difference in the plasma concentration of folate, cobalamin, or methylmalonic acid between the 2 groups. Patients whose homocysteine levels were in the higher quartile (>or=17.7 micro mol/L) had increased prevalence of CAD (relative risk, 1.75; 95% confidence interval, 1.08-2.70;P=.04). CONCLUSIONS: Levodopa therapy, rather than PD, is a cause of hyperhomocysteinemia in patients with PD. Deficiency of folate or vitamin B(12) levels does not explain the elevated homocysteine levels in these patients. To our knowledge, this is the first report that levodopa-related hyperhomocysteinemia is associated with increased risk for CAD. These findings have implications for the treatment of PD in patients at risk for vascular disease, and potentially for those at risk for dementia and depression. PMID- 12533090 TI - Lower levels of N-acetylaspartate in multiple sclerosis patients with the apolipoprotein E epsilon4 allele. AB - BACKGROUND: In multiple sclerosis (MS), the epsilon4 allele of apolipoprotein E (APOE epsilon4) has been associated with more rapid clinical worsening and more severe tissue damage on magnetic resonance imaging. OBJECTIVE: To use proton magnetic resonance spectroscopy ((1)H-MRS) to further explore the biochemical changes in the brains of patients with MS associated with APOE epsilon4. DESIGN: A 2-year clinical and (1)H-MRS follow-up cohort study. SETTING: The MS outpatient clinic, Department of Neurology, and Magnetic Resonance Center of Karl-Franzens University. PATIENTS: We performed (1)H-MRS of the central portion of both hemispheres and APOE genotyping in 72 patients (52 women and 20 men; mean +/- SD age, 34.8 +/- 8.8 years) with clinically definite relapsing-remitting MS. Repeated studies were performed in 44 patients after a mean +/- SD interval of 34 +/- 9 months. MAIN OUTCOME MEASURE: Levels of N-acetylaspartate as measured by (1)H-MRS. RESULTS: Patients with MS and an epsilon4 allele (n = 19) had a significantly lower mean +/- SD N-acetylaspartate-creatine ratio than those without an epsilon4 allele (n = 53) (1.73 +/- 0.26 vs 1.89 +/- 0.24; P =.04) despite the absence of significant differences in age at onset, disease duration, Expanded Disability Status Scale score, and number of previous relapses between subgroups. During follow-up, the drop in the N-acetylaspartate-creatine ratio of epsilon4 carriers was also significantly larger (-0.31 vs -0.10; P =.01). This was paralleled by a higher number of relapses (mean +/- SD, 4.1 +/- 2.7 vs 1.7 +/ 1.6; P =.02) and a faster although nonsignificant progression of disability (mean +/- SD (Delta)Expanded Disability Status Scale score, 0.9 +/- 1.8 vs 0.3 +/ 1.1; P =.19). CONCLUSIONS: The APOE epsilon4 allele has a negative effect on the course of MS, and increasing axonal damage may be an important mechanism. PMID- 12533091 TI - Drop attacks and vertigo secondary to a non-meniere otologic cause. AB - BACKGROUND: Tumarkin falls are sudden drop-attack falls that occur in a subset of patients with Meniere syndrome (endolymphatic hydrops), an inner ear disorder characterized by vertigo spells and hearing loss. OBJECTIVE: To describe the clinical features and quantitative audiovestibular testing results in a case series of patients with Tumarkin falls, episodic vertigo, and normal hearing. SETTING: University referral center for disorders of balance and hearing. METHODS: Case series (unselected) of all patients with Tumarkin falls and a normal audiogram at least 1 year after onset of vestibular symptoms (n = 6) from a retrospective analysis of the records of all patients with Tumarkin falls presenting to Neurotology Clinic at UCLA Medical Center, Los Angeles, Calif, from October 1, 1975, to February 1, 2001 (N = 55). Quantitative audiologic and vestibular function testing, neurologic history, and examination were performed. RESULTS: Five of 6 patients had unilateral caloric paresis, and 1 had bilateral vestibulopathy. Five of 6 had a personal and/or family history of migraine headaches meeting International Headache Society criteria. All patients had a subjective sensation of feeling pushed by an external force, and half of the patients had a subjective tilt of the environment concurrent with the fall. CONCLUSIONS: The incidence of migraine is high in this subgroup of patients with Tumarkin falls and normal hearing. The clinical description of the falls is similar to those associated with Meniere syndrome. Further studies are needed to understand the etiology of Tumarkin falls in these patients with normal hearing. PMID- 12533092 TI - Effects of subthalamic nucleus stimulation and levodopa treatment on gait abnormalities in Parkinson disease. AB - BACKGROUND: Stimulation of the subthalamic nucleus is proposed for the treatment of patients presenting with severe Parkinson disease. The effect on gait is not clearly established. OBJECTIVES: To evaluate objectively the influence of bilateral subthalamic nucleus stimulation on gait in Parkinson disease and to compare it with the effects of levodopa treatment. METHODS: Ten patients underwent bilateral subthalamic nucleus stimulation. The preoperative and postoperative (3 months after surgery) clinical gait disturbances, as well as spatial and temporal gait parameters, were analyzed in off and on-drug conditions. The gait analysis was performed using a video motion analysis system (optoelectronic VICON system; Oxford Metrics, Oxford, England). RESULTS: In the off condition, there was an improvement after surgery for the total motor score and the gait subscore. In the on-drug condition, there was an improvement in levodopa-induced dyskinesias and the motor score, whereas the gait subscore was unchanged. For the gait parameters measured by the video motion analysis system system, there was also an improvement in the off condition and to a lesser extent in the on-drug condition. CONCLUSIONS: Our method allowed exact quantification of the benefit of surgery on gait parameters. Compared with the levodopa treatment, the effect of stimulation on gait kinematic parameters seems to be qualitatively similar but quantitatively different with a lower benefit on gait velocity and stride length. Concerning the pathophysiology of gait troubles in Parkinson disease, the deficit in control of stride length would be the fundamental deficit. The study underlines the possible role of the subthalamic nucleus on the stride length regulation. PMID- 12533093 TI - Survival study of vascular dementia in Rochester, Minnesota. AB - OBJECTIVE: To investigate the relationship between features and definitions of vascular dementia (VaD) and survival. DESIGN: We used the medical records linkage system of the Rochester Epidemiology Project to identify incident cases of dementia in Rochester from January 1, 1985, through December 31, 1989. Dementia and Alzheimer disease were defined using the criteria of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition. Vascular dementia was defined by ad hoc criteria, including imaging. Each patient with dementia was matched by age and sex to a referent subject free of dementia. Patients with dementia and referent subjects were followed from the onset of dementia (or index year) through death, censoring, or the end of the study. RESULTS: We included 479 patients with incident dementia and 479 referent subjects. Overall, patients with VaD had worse mortality than referent subjects (relative risk [RR], 2.7; 95% confidence interval [CI], 1.9-3.9). Among patients with VaD, those with dementia temporally related to a stroke had a worse relative mortality (RR, 4.5; 95% CI, 2.7-7.4) than those with only imaging evidence of bilateral infarctions in gray matter structures (RR, 2.4; 95% CI, 1.5-3.8). Relative mortality estimates varied by using 3 sets of published diagnostic criteria for VaD. Patients with VaD had a higher RR of death (RR, 2.7; 95% CI, 1.9-3.9) than patients with dementia overall (RR, 1.8; 95% CI, 1.6-2.1) or patients with Alzheimer disease (RR, 1.4; 95% CI, 1.2-1.7). CONCLUSIONS: The relative mortality of patients with VaD varied depending on the set of diagnostic criteria used. A temporal relationship to a stroke was the strongest predictive feature for poor survival in patients with dementia. PMID- 12533094 TI - Survival study of Parkinson disease in Olmsted County, Minnesota. AB - OBJECTIVE: To compare survival in incident cases of Parkinson disease (PD) with survival in subjects free of PD from the general population. METHODS: We used the medical records linkage system of the Rochester Epidemiology Project to identify incident cases of PD in Olmsted County, Minnesota, for the period 1976-1995. Cases were matched by age and sex to referent subjects from the same population. For 196 cases and 185 referent subjects, we studied survival between the date of diagnosis of PD (or index date) and death, loss to follow-up, or end of the study (May 1, 2000). RESULTS: The median length of follow-up was 7.2 years for cases and 8.0 years for referent subjects; 110 patients with PD and 79 referent subjects died during follow-up. The median survival was 10.3 years in cases and 13.4 years in referent subjects. The relative risk (RR) of death was 1.60 (95% confidence interval [CI], 1.20-2.14; P =.002) overall, 1.81 (95% CI, 1.15-2.84; P =.01) in women, and 1.49 (95% CI, 1.01-2.20; P =.04) in men. There was a decreasing trend in the RR of death according to age at onset of PD (in tertiles): younger than 67 years, RR, 2.04 (95% CI, 0.99-4.19; P =.05); 67 to 76 years, RR, 1.76 (95% CI, 1.08-2.86; P =.02); and older than 76 years, RR, 1.48 (95% CI, 0.95-2.29; P =.08). Patients with PD who had both rest tremor and pronounced asymmetry had a better prognosis than patients with neither clinical characteristic. Patients with PD who smoked survived better than expected. CONCLUSIONS: Patients with PD face a higher risk of death compared with subjects free of PD from the general population. Certain clinical characteristics and smoking modify survival. PMID- 12533095 TI - A novel central nervous system-enriched spinocerebellar ataxia type 7 gene product. AB - CONTEXT: Polyglutamine-mediated neurodegeneration in spinocerebellar ataxia type 7 (SCA7) involves specific central nervous system structures despite widespread expression of the mutant ataxin-7 protein. OBJECTIVE: To determine whether expression of multiple gene products could contribute to selective neurodegeneration in SCA7. RESULTS: We identified a novel SCA7 transcript and protein, both of which are enriched within the central nervous system. An isoform specific antibody revealed that the novel ataxin-7 variant, in contrast with the previously described protein, localizes to neuronal cytoplasm and not to inclusion bodies present within the tissues of patients with SCA7. CONCLUSIONS: In addition to expanding our understanding of SCA7 gene expression, identification of a novel ataxin-7 protein enriched in the central nervous system suggests that expression of multiple polyglutamine-containing proteins may play a role in generating the neurodegenerative patterns characteristic of SCA7 and other polyglutamine expansion diseases. PMID- 12533096 TI - A fatal case of coxsackievirus B4 meningoencephalitis. AB - BACKGROUND: Coxsackieviruses and echoviruses are common causes of aseptic meningitis, but they rarely cause life-threatening illness. We report a fatal case of coxsackievirus B4 meningoencephalitis in a woman who developed extrapyramidal symptoms suggestive of encephalitis lethargica. The exact causative agent of encephalitis lethargica has rarely been found, but most cases of the syndrome are assumed to be of viral origin. CASE DESCRIPTION: A 33-year old woman previously treated with methylprednisolone and cyclophosphamide for Henoch-Schonlein purpura was transferred from a referring hospital because of sore throat, fever, and chills. Her neurologic findings progressed from headache with mild photophobia to lethargy, cogwheeling, increased tone in all 4 limbs, and brisk reflexes. The patient was diagnosed as having coxsackievirus B4 meningoencephalitis and, despite treatment with the experimental antiviral agent pleconaril, died of an overwhelming central nervous system infection and myocarditis. Magnetic resonance imaging showed focal hyperintense lesions in the substantia nigra that corresponded to the location of pathological changes seen at autopsy. CONCLUSIONS: This patient had a fulminant coxsackievirus B4 viral meningoencephalitis with a clinical pattern reminiscent of encephalitis lethargica and striking focal abnormalities in the substantia nigra identified on magnetic resonance imaging. The magnetic resonance imaging findings correlated with pathological changes identified at autopsy that were similar to the pathological findings observed in patients with encephalitis lethargica and postencephalitic parkinsonism. It is likely that the patient's immunocompromised state led to an overwhelming infection from an otherwise relatively innocuous viral infection. PMID- 12533097 TI - Hearing symptoms in migrainous infarction. AB - BACKGROUND: In case reports, migraine headaches have been associated with fluctuating low-frequency hearing loss and sudden, unilateral hearing loss. Auditory symptoms associated with migrainous infarction have not previously been emphasized. OBJECTIVE: To describe migrainous infarction presenting with acute auditory symptoms. DESIGN: Case reports. SETTING: Tertiary care hospitals. PATIENTS: A 40-year-old man with a history of migraine suddenly developed bilateral hearing loss associated with severe, throbbing, occipital headache, tinnitus, vertigo, speech disturbance, and right hemiparesis. An early audiogram showed profound, down-sloping, sensorineural-type hearing loss bilaterally. Sixteen days later, a follow-up pure tone audiogram documented marked improvement in both sides to a pure tone average of 30 dB. Right hemiparesis and dysarthria also improved steadily for 2 months. A 25-year-old woman with a history of migraine with aura suddenly developed hyperacusis, unilateral hearing loss, and migraine headache early in migrainous infarction. Magnetic resonance imaging documented infarcts in the pons and cerebellum. CONCLUSIONS: In these patients, acute auditory symptoms are a part of the prodrome of migrainous infarction. We speculate that these symptoms may have resulted from migraine-associated vasospasm. Migrainous infarction should be considered in the differential diagnosis of acute auditory symptoms, including sudden, bilateral hearing loss. PMID- 12533098 TI - The fragile X premutation presenting as essential tremor. AB - CONTEXT: The fragile X premutation has recently been reported to be associated with a neurodegenerative syndrome, chiefly characterized by intention tremor, gait ataxia, and executive cognitive deficits in men older than 50 years. Essential tremor is a frequent cause of tremor in elderly patients and in some cases is associated with impaired tandem gait and cognitive deficits. OBJECTIVE: To describe 2 fragile X carriers whose clinical presentation mimicked essential tremor. DESIGN: The 2 patients described herein underwent neurologic examinations by experienced movement disorders neurologists, magnetic resonance imaging, and fragile X gene, messenger RNA, and protein analyses. One underwent detailed neuropsychological testing. SETTING: Patients were studied at 2 large university movement disorders clinics. PATIENTS: Both patients were white men older than 50 years who had been diagnosed as having essential tremor and then found to be fragile X carriers. RESULTS: Besides disabling intention tremor, the 2 patients had impaired tandem, generalized brain atrophy, and unusual bilateral T2 middle cerebellar hyperintensities on magnetic resonance imaging. The patient who underwent neuropsychological testing had frontal executive deficits. Both patients had elevated fragile X mental retardation gene 1 messenger RNA and reduced fragile X mental retardation 1 protein levels. CONCLUSION: The fragile X carrier state may underlie the clinical findings in some older men diagnosed as having essential tremor. PMID- 12533099 TI - Spinal arteriovenous malformation. PMID- 12533100 TI - The history of dermatome mapping. AB - Dermatome maps are commonly used in clinical neurology. These maps are valuable for the localization of varied sensory phenomena in patients with neurological disorders. The methods used in the construction of the classic maps by Sir Henry Head, Sir Charles Sherrington, Otfrid Foerster, and Jay Keegan and Frederic Garrett are of historical interest and are relevant to the current understanding of dermatome anatomy and physiology. In particular, the work of Derek Denny-Brown and his colleagues demonstrates that patterns of dermatomal sensory loss depend on the anatomical and physiological characteristics of large regions of nervous tissue, multiple adjacent dorsal ganglia, and the nearby caudal and rostral spinal cord. PMID- 12533101 TI - Are astrocytes central players in the pathophysiology of multiple sclerosis? AB - An interaction between antimyelin T cells and antigen-presenting glial cells is a crucial step in the cascade of immune events that lead to the inflammatory lesions in multiple sclerosis (MS). One of the most debated and controversial issues is whether microglial cells or astrocytes are the key players in initiating the (auto)immune reactions in the central nervous system in MS. Many investigators consider microglia to be the responsible intrinsic immunoeffector cells. In this review, we speculate that in MS astrocytes may serve as primary (facultative) antigen-presenting cells due to a failure of noradrenergic suppression of class II major histocompatibility complex molecules, which is caused by a loss of beta(2)-adrenergic receptors. If this hypothesis is correct, pharmacologic suppression of the antigen-presenting capacities of astrocytes may be a potential therapy for MS. PMID- 12533103 TI - Specialty hospitals generate revenue and controversy. PMID- 12533104 TI - David Meltzer, physician and economist, discusses the new hospitalist movement. Interview by Sarah Pressman Lovinger. PMID- 12533102 TI - Sex and gender factors in medical studies: implications for health and clinical practice. PMID- 12533105 TI - Soccer "sudden death" given new meaning. PMID- 12533106 TI - Exercise intensity and risk of coronary heart disease. PMID- 12533107 TI - Exercise intensity and risk of coronary heart disease. PMID- 12533109 TI - Splinting vs surgery for carpal tunnel syndrome. PMID- 12533111 TI - Splinting vs surgery for carpal tunnel syndrome. PMID- 12533113 TI - Splinting vs surgery for carpal tunnel syndrome. PMID- 12533114 TI - Splinting vs surgery for carpal tunnel syndrome. PMID- 12533115 TI - Splinting vs surgery for carpal tunnel syndrome. PMID- 12533116 TI - Splinting vs surgery for carpal tunnel syndrome. PMID- 12533117 TI - Financial consequences of drug benefit plans. PMID- 12533118 TI - Financial consequences of drug benefit plans. PMID- 12533119 TI - Financial consequences of drug benefit plans. PMID- 12533121 TI - Escalation of drug use in early-onset cannabis users vs co-twin controls. AB - CONTEXT: Previous studies have reported that early initiation of cannabis (marijuana) use is a significant risk factor for other drug use and drug-related problems. OBJECTIVE: To examine whether the association between early cannabis use and subsequent progression to use of other drugs and drug abuse/dependence persists after controlling for genetic and shared environmental influences. DESIGN: Cross-sectional survey conducted in 1996-2000 among an Australian national volunteer sample of 311 young adult (median age, 30 years) monozygotic and dizygotic same-sex twin pairs discordant for early cannabis use (before age 17 years). MAIN OUTCOME MEASURES: Self-reported subsequent nonmedical use of prescription sedatives, hallucinogens, cocaine/other stimulants, and opioids; abuse or dependence on these drugs (including cannabis abuse/dependence); and alcohol dependence. RESULTS: Individuals who used cannabis by age 17 years had odds of other drug use, alcohol dependence, and drug abuse/dependence that were 2.1 to 5.2 times higher than those of their co-twin, who did not use cannabis before age 17 years. Controlling for known risk factors (early-onset alcohol or tobacco use, parental conflict/separation, childhood sexual abuse, conduct disorder, major depression, and social anxiety) had only negligible effects on these results. These associations did not differ significantly between monozygotic and dizygotic twins. CONCLUSIONS: Associations between early cannabis use and later drug use and abuse/dependence cannot solely be explained by common predisposing genetic or shared environmental factors. The association may arise from the effects of the peer and social context within which cannabis is used and obtained. In particular, early access to and use of cannabis may reduce perceived barriers against the use of other illegal drugs and provide access to these drugs. PMID- 12533122 TI - External incentives, information technology, and organized processes to improve health care quality for patients with chronic diseases. AB - CONTEXT: Organized care management processes (CMPs) can improve health care quality for patients with chronic diseases. The Institute of Medicine of the National Academy of Sciences has called for public and private purchasers of health care to create incentives for physician organizations (POs) to use CMPs and for the government to assist POs in implementing information technology (IT) to facilitate CMP use. Research is lacking about the extent to which POs use CMPs or about the degree to which incentives, IT, or other factors are associated with their use. OBJECTIVES: To determine the extent to which POs with 20 or more physicians use CMPs and to identify key factors associated with CMP use for 4 chronic diseases (asthma, congestive heart failure, depression, and diabetes). DESIGN, SETTING, AND PARTICIPANTS: One thousand five hundred eighty-seven US POs (medical groups and independent practice associations) with 20 or more physicians were identified using 5 large databases. One thousand one hundred four of these POs (70%) agreed to participate in a telephone survey conducted between September 2000 and September 2001. Sixty-four responding POs were excluded because they did not treat any of the 4 diseases, leaving 1040 POs. MAIN OUTCOME MEASURES: Extent of use of CMPs as calculated on the basis of a summary measure, a PO care management index (POCMI; range, 0-6) and factors associated with CMP use. RESULTS: Physician organizations' mean use of CMPs was 5.1 of a possible 16; 50% used 4 or fewer. External incentives and clinical IT were most strongly associated with CMP use. Controlling for other factors, use of the 2 most strongly associated incentives-public recognition and better contracts for health care quality-was associated with use of 1.3 and 0.7 additional CMPs, respectively (P<.001 and P =.007). Each additional IT capability was associated with 0.37 additional CMPs (P<.001). However, 33% of POs reported no external incentives and 50% reported no clinical IT capability. CONCLUSIONS: The use of CMPs varies greatly among POs, but it is low on average. Government and private purchasers of health care may increase CMP use by providing external incentives for improvement of health care quality to POs and by assisting them in improving their clinical IT capability. PMID- 12533123 TI - Changes in career satisfaction among primary care and specialist physicians, 1997 2001. AB - CONTEXT: A number of forces have changed the practice of medicine in the past decade. Evidence suggests that physicians are becoming less satisfied in this environment. OBJECTIVES: To describe changes in career satisfaction in a large, nationally representative sample of physicians and to examine market and practice factors associated with changes in physician satisfaction. DESIGN AND SETTING: Data were collected from the first 3 rounds of the Community Tracking Study (CTS) Physician Survey, a series of nationally representative telephone surveys of physicians in 60 US sites conducted in 1996-1997 (round 1: 12 385 respondents; 65% response rate), 1998-1999 (round 2: 12 280 respondents; 61% response rate), and 2000-2001 (round 3: 12 389 respondents; 59% response rate) for the Center for Studying Health System Change. The second and third rounds of the survey included physicians sampled in the previous round, as well as new physicians. PARTICIPANTS: Primary care and specialist physicians who spent at least 20 hours per week in direct patient care activities. MAIN OUTCOME MEASURES: Changes in physicians' overall satisfaction with their career and the proportion of dissatisfied physicians in particular sites. RESULTS: Physician satisfaction levels declined marginally between 1997 and 2001, with most of the decline occurring between 1997 and 1999. Among primary care physicians, 42.4% were very satisfied in 1997, as were 43.3% of specialists, compared with 38.5% and 41.4%, respectively, in 2001. There were nearly equal increases in those who reported that they were somewhat satisfied. Overall means mask significant differences across the 60 sites. Among 12 sites randomly selected for more intensive study, the proportion of respondents who were somewhat or very dissatisfied ranged from 8.8% of physicians in Lansing, Mich (1999), to 34.2% in Miami, Fla (1997). Between 1997 and 1999, 25.6% of primary care physicians reported decreased satisfaction and 18.1% reported improved satisfaction, while approximately equal percentages reported increased (19.8%) and decreased (20.4%) satisfaction between 1999-2001. Findings were similar for specialist physicians. In multivariable models, the strongest and most consistent predictors of change in satisfaction were changes in measures of clinical autonomy, including increases in hours worked and physicians' ability to obtain services for their patients. Changes in exposure to managed care were weakly related to changes in satisfaction. CONCLUSIONS: Our findings demonstrate that overall physician satisfaction levels over this time period did not change dramatically. In addition, satisfaction and changes in satisfaction vary greatly among sites. Rather than declining income, threats to physicians' autonomy, to their ability to manage their day-to-day patient interactions and their time, and to their ability to provide high-quality care are most strongly associated with changes in satisfaction. PMID- 12533124 TI - Patterns and trends in food portion sizes, 1977-1998. AB - CONTEXT: While general consensus holds that food portion sizes are increasing, no empirical data have documented actual increases. OBJECTIVE: To determine trends in food portion sizes consumed in the United States, by eating location and food source. DESIGN, SETTING, AND PARTICIPANTS: Nationally representative data from the Nationwide Food Consumption Survey (1977-1978) and the Continuing Survey of Food Intake by Individuals (1989-1991, 1994-1996, and 1998). The sample consists of 63 380 individuals aged 2 years and older. MAIN OUTCOME MEASURE: For each survey year, average portion size consumed from specific food items (salty snacks, desserts, soft drinks, fruit drinks, french fries, hamburgers, cheeseburgers, pizza, and Mexican food) by eating location (home, restaurant, or fast food). RESULTS: Portion sizes vary by food source, with the largest portions consumed at fast food establishments and the smallest at other restaurants. Between 1977 and 1996, food portion sizes increased both inside and outside the home for all categories except pizza. The energy intake and portion size of salty snacks increased by 93 kcal (from 1.0 to 1.6 oz [28.4 to 45.4 g]), soft drinks by 49 kcal (13.1 to 19.9 fl oz [387.4 to 588.4 mL]), hamburgers by 97 kcal (5.7 to 7.0 oz [161.6 to 198.4 g]), french fries by 68 kcal (3.1 to 3.6 oz [87.9 to 102.1 g]), and Mexican food by 133 kcal (6.3 to 8.0 oz [178.6 to 226.8 g]). CONCLUSION: Portion sizes and energy intake for specific food types have increased markedly with greatest increases for food consumed at fast food establishments and in the home. PMID- 12533125 TI - Scope and impact of financial conflicts of interest in biomedical research: a systematic review. AB - CONTEXT: Despite increasing awareness about the potential impact of financial conflicts of interest on biomedical research, no comprehensive synthesis of the body of evidence relating to financial conflicts of interest has been performed. OBJECTIVE: To review original, quantitative studies on the extent, impact, and management of financial conflicts of interest in biomedical research. DATA SOURCES: Studies were identified by searching MEDLINE (January 1980-October 2002), the Web of Science citation database, references of articles, letters, commentaries, editorials, and books and by contacting experts. STUDY SELECTION: All English-language studies containing original, quantitative data on financial relationships among industry, scientific investigators, and academic institutions were included. A total of 1664 citations were screened, 144 potentially eligible full articles were retrieved, and 37 studies met our inclusion criteria. DATA EXTRACTION: One investigator (J.E.B.) extracted data from each of the 37 studies. The main outcomes were the prevalence of specific types of industry relationships, the relation between industry sponsorship and study outcome or investigator behavior, and the process for disclosure, review, and management of financial conflicts of interest. DATA SYNTHESIS: Approximately one fourth of investigators have industry affiliations, and roughly two thirds of academic institutions hold equity in start-ups that sponsor research performed at the same institutions. Eight articles, which together evaluated 1140 original studies, assessed the relation between industry sponsorship and outcome in original research. Aggregating the results of these articles showed a statistically significant association between industry sponsorship and pro-industry conclusions (pooled Mantel-Haenszel odds ratio, 3.60; 95% confidence interval, 2.63-4.91). Industry sponsorship was also associated with restrictions on publication and data sharing. The approach to managing financial conflicts varied substantially across academic institutions and peer-reviewed journals. CONCLUSIONS: Financial relationships among industry, scientific investigators, and academic institutions are widespread. Conflicts of interest arising from these ties can influence biomedical research in important ways. PMID- 12533126 TI - A 59-year-old woman with gastroesophageal reflux disease and Barrett esophagus. PMID- 12533127 TI - A 52-year-old man with a positive PPD, 1 year later. PMID- 12533128 TI - Direct-to-consumer advertising and shared liability for pharmaceutical manufacturers. PMID- 12533129 TI - Does marijuana use cause the use of other drugs? PMID- 12533132 TI - Some pioneers in plastic surgery of the facial region. PMID- 12533133 TI - Development of the upper lip. AB - OBJECTIVES: To affirm and reanalyze George L. Streeter's "merging theory" of upper-lip development in primates by observing progressive embryologic stages in facial development using scanning electron microscopy (SEM) and to further understand upper-lip development. DESIGN: The study was conducted at the California Regional Primate Research Center, Davis. Twenty primate embryos (Macaca fascicularis) and 2 fetuses were examined with SEM. The development of the frontonasal prominence, maxillary prominence, medial nasal prominence, and lateral nasal prominence were sequentially observed. The contribution of these prominences to the formation of the upper lip and nose were carefully analyzed. RESULTS: The maxillary prominence and medial nasal prominence form the upper lip, whereas the lateral nasal, medial nasal, and maxillary prominences form the nose. There is fusion of the maxillary prominence with the medial nasal prominence. This fusion has not been previously described. This has resulted in a modification of the current theory of upper-lip development into one we refer to as the "dynamic fusion theory." CONCLUSIONS: The dynamic fusion theory explains the merging process of the mesenchymal and ecotodermal layers of the facial prominences that contribute to the upper-lip formation. The dynamic fusion theory of facial prominence movement details the interaction between epithelial layers: both epithelial layers must fuse properly to avoid cleft-lip deformities. PMID- 12533134 TI - Autocrine growth factor production by fetal, keloid, and normal dermal fibroblasts. AB - OBJECTIVE: To evaluate differences in fibroblast autocrine growth factor production by human fetal, keloid, and normal adult dermal fibroblasts. DESIGN: Serum-free cell lines of fetal, keloid, and normal adult dermal fibroblasts were established. Cell counts were performed and supernatants collected at 4, 24, and 72 hours. Cell-free supernatants were quantitatively assayed for transforming growth factor beta1 (TGF-beta1) and basic fibroblast growth factor (bFGF). RESULTS: Population doubling times for fetal, keloid, and normal adult fibroblasts were 120.0, 88.1, and 128.4 hours, respectively. Differences in population doubling times did not reach statistical significance. Statistically significant differences between TGF-beta1 levels from fetal and normal adult fibroblasts were seen at 24 and 72 hours. Significant differences between TGF beta1 levels from keloid and normal adult fibroblasts were also seen at 24 and 72 hours. Fetal fibroblasts demonstrated higher levels of bFGF than normal adult fibroblasts at each time point, but these differences were not statistically significant. No significant differences were observed between keloid and normal adult bFGF levels. CONCLUSIONS: Both fetal and keloid fibroblasts produce significantly more TGF-beta1 than normal adult fibroblasts. Our data and the data of others suggest that fetal fibroblasts produce more bFGF than adult fibroblasts. The serum-free model we describe can be used to quantitatively measure autocrine growth factor production by cells that underlie clinically different types of wound healing. This model provides information that may allow us to better treat and prevent undesirable scarring. PMID- 12533135 TI - Inhibitors of nitric oxide promote microvascular thrombosis. AB - BACKGROUND: Microvascular free tissue transfer is a widely utilized method of head and neck reconstruction. Despite advances in the field, reports of experienced microvascular surgeons on large series of free flap procedures reveal that the incidence of free flap failure varies between 5% and 9%. Most cases of free flap failure are initiated by platelet-mediated events that result in thrombosis at the microvascular anastomoses. Recent evidence indicates that nitric oxide (NO) plays a critical role in preventing thrombosis by inhibiting platelet adhesion and aggregation. The role of NO in microvascular anastomotic thrombosis has not been studied. OBJECTIVE: To determine the role of NO in microvascular thrombosis using an in vivo rabbit model. METHODS: An arterial inversion graft (AIG)-induced microvascular thrombosis model was utilized in New Zealand white rabbits. The femoral arteries were used bilaterally to create 3-mm AIGs. Intravenous NO donor, NO inhibitor, or isotonic sodium chloride solution (control) was administered for 1 hour following the completion of the AIG, and vessel patency was then checked using a direct "milking test." Sixteen rabbits (32 AIGs) were used as controls. A potent NO inhibitor, N(w)-nitro-L-arginine methylester (L-NAME), was administered to 13 rabbits (26 AIGs) and L-arginine, a NO precursor/donor, was given to 10 rabbits (20 AIGs). RESULTS: The control animals had a thrombosis rate of 46.9%. The rate of thrombosis in animals exposed to an NO inhibitor (L-NAME) was significantly higher, at 76.9% (P<.05, chi( 2) = 4.23). The L-arginine group did not show a statistical difference with the control in the rate of thrombosis (50.0%). CONCLUSIONS: Nitric oxide plays a role in microvascular anastamotic thrombosis. Intravenous NO inhibitors appear to increase the short-term rate of microvascular thrombosis. L-arginine, an NO precursor, does not appear to produce the opposite effect. Further studies using local NO donors and antagonists as well as more potent NO precursors are needed to further evaluate NO's role in microvascular thrombosis. The results of this study may have applications to human microvascular surgery. PMID- 12533136 TI - The effect of subcutaneously administered low-molecular-weight heparin on microarterial thrombosis in the rat. AB - OBJECTIVE: To examine the effect of administration of a low-molecular-weight heparin derivative, enoxaparin, on the rate of arterial thrombosis in a rat model. STUDY DESIGN: Prospective, randomized, blinded study. METHODS: A standard microarterial anastomosis tuck injury was created in both femoral arteries of 25 Long Evans retired breeder rats. Thirteen animals received a subcutaneous injection of 50 IU/kg of enoxaparin 2 hours before the procedure, while 12 control animals received vehicle (isotonic sodium chloride solution) alone. Sites of injury/repair were assessed 2 hours after the procedure for anastomotic patency or thrombosis. RESULTS: Six (23%) of 26 vessels in the drug-treated group developed an arterial thrombosis at the site of repair, while 6 (25%) of 24 vessels in the control group developed thrombosis. There was no statistically significant difference at the 95% confidence limit between the 2 groups based on a comparison-of-proportions test. CONCLUSION: The preoperative subcutaneous administration of 50 IU/kg of enoxaparin did not alter the rate of arterial thrombosis following the creation of a thrombogenic tuck injury/repair of the rat femoral artery. PMID- 12533137 TI - Decellularized dermal grafting in cleft palate repair. AB - OBJECTIVE: To assess the efficacy of decellularized dermal grafting used as an adjunct to the performance of primary repair of wide cleft palates. DESIGN: Retrospective review. SETTING: Tertiary referral center for large managed care organization. METHODS: Seven consecutive patients with clefts of the hard and soft palates wider than 15 mm as measured at the posterior edge of the hard palate. Palates were repaired in the standard 2-flap approach with intravelar veloplasty. The decellularized dermal graft (AlloDerm) was applied immediately deep to the oral mucosal closure. Patients were followed up with serial postoperative examination. Palates were assessed for dehiscence, fistula, infection, rejection, scarring, and contracture. RESULTS: There were no fistulas. In 2 patients, the oral mucosa dehisced, exposing the dermal graft. In 2 other cases, nasal mucosal tears were inadvertently created during closure of the nasal layer. In all cases, the decellularized dermal graft mucosalized and, by clinical examination, became incorporated into the wound. There were no cases of local inflammation or infection. The degree of scarring and contracture was indistinguishable from the adjacent scar. CONCLUSIONS: Decellularized dermal graft is safe and effective for use in primary closure of wide clefts involving the hard and soft palates. Its application to wide clefts otherwise at risk of fistula is justified. Its use in repair of an existing fistula is also promising. PMID- 12533139 TI - Radiofrequency cartilage reshaping: efficacy, biophysical measurements, and tissue viability. AB - OBJECTIVES: To assess the feasibility of reshaping cartilage using radiofrequency (RF) heating, and to examine the effects of this process on tissue biophysical properties (optical and thermal) and cellular viability. METHODS: Mechanically deformed porcine septal cartilage was reshaped using 2 RF-generating devices. We performed dynamic measurements of tissue thermal and optical properties while heating cartilage with one of these devices. Cellular viability was assessed immediately and 7 days after treatment. RESULTS: A characteristic change in the diffuse transmittance of light through the cartilage occurred during heating. Change in transmittance has been shown to accompany the onset of stress relaxation in cartilage. Peak radiometric surface temperature during heating was 88.6 degrees C. Specimens retained their user-specified curved shape for the observed period of 14 days. Chondrocyte viability in RF-heated tissue was 19% and 14% of that in untreated control specimens at days 0 and 7 after treatment, respectively. CONCLUSIONS: Radiofrequency heating has been shown to effectively reshape cartilage while maintaining cellular viability, illustrating a novel application for a widely used technology. PMID- 12533141 TI - Effect of topical mitomycin on skin wound contraction. AB - OBJECTIVE: To evaluate the effect of different dosing regimens of mitomycin on skin wound contraction. METHODS: Full-thickness skin wounds were created in 5 groups of hairless mice, which represented different dosing regimens or a sterile water control: A, control; B, mitomycin (0.5 mg/mL) applied immediately after creation of the lesion (day 1); C, mitomycin (1.0 mg/mL) applied on day 1; D, mitomycin (0.5 mg/mL) applied on days 1 and 3; and E, mitomycin (1.0 mg/mL) applied on days 1 and 3. Wound surface area was measured immediately after drug application (day 1), and thereafter every 3 to 5 days until day 29 by means of computer-assisted image analysis. RESULTS: All dosing regimens of mitomycin application resulted in an initially exponential rate of wound contraction that was significantly slower than in the sterile water control group, with a significantly larger wound surface area on day 29. Wound area in the control group contracted approximately 9 times more rapidly than in the treatment groups. No difference was observed among the different dosing regimens. CONCLUSION: Application of mitomycin, at the lowest dose and frequency of application used in this study, resulted in improved outcomes with regard to contraction of full thickness skin wounds. PMID- 12533140 TI - A compositional analysis of human nasal septal cartilage. AB - BACKGROUND: Nasal septal cartilage is well established as an autograft material. Tissue engineering methods are now being developed to synthesize cartilage constructs with the properties of this type of cartilage. However, important baseline data on the composition of native septal cartilage is sparse. OBJECTIVES: To characterize quantitatively the major biochemical constituents of native adult human septal cartilage and determine age- or sex-related variation in composition. METHODS: Cartilage was harvested from the inferior region of the nasal septum in 33 patients (mean +/- SD age, 47.0 +/- 13.5 years; range, 24-80 years) during routine septoplasty or septorhinoplasty. Biochemical assays were used to determine the quantities, relative to wet weight, of the major constituents of cartilage: water, collagen (from hydroxyproline), sulfated glycosaminoglycan (sGAG), and chondrocytes (from DNA). RESULTS: On average, each gram of wet cartilage contained 77.7% water, 7.7% collagen, 2.9% sGAG, and 24.9 million cells. Hydration and collagen content showed no significant age variation. Advancing age was associated with a reduction in sGAG content (7.7% per decade, P =.02) and cellularity (7.4% per decade, P =.05). No significant sex differences were found in any of these cartilage constituents. CONCLUSIONS: This study represents the first biochemical characterization of the composition of native human septal cartilage. The data serve as a baseline for future comparison of the properties of tissue-engineered neocartilage constructs. Furthermore, the age-associated variations in cartilage composition have implications for patient selection for reconstructive procedures. PMID- 12533142 TI - Strength and histological characteristics of periosteal fixation to bone after elevation. AB - BACKGROUND: Loss of brow elevation in the early postoperative period has been well documented; however, there has been no study quantifying the minimum time necessary to achieve lasting brow elevation. Previous work in our laboratory has demonstrated that complete readherence of periosteum to bone can take 12 weeks to occur after periosteal elevation. The correlation between adherence and the development of strength has never been examined. OBJECTIVE: To correlate histological characteristics of raised forehead periosteum with the strength of the periosteum-bone union postoperatively. SUBJECTS: Eighteen New Zealand white rabbits. METHODS: Rabbit foreheads were elevated in a subperiosteal plane and the flap reapproximated with chromic sutures. Animals were killed at 14, 28, 45, 63, or 84 days postoperatively, and all tissue superficial to the periosteum removed. The tension required to avulse sections of periosteum was then measured. Skulls were then sectioned and prepared for histological analysis of remaining periosteum. Avulsion forces and histological findings were compared with those unoperated-on controls. RESULTS: The forces necessary to avulse periosteum in the 14- and 28-day groups were significantly lower than for control animals; values at 45, 63, and 84 days were not significantly different from control animals. Healing periosteum displayed varying degrees of thickness, cellularity, edema, and vascular congestion. These features peaked at 28 days postoperatively then gradually resolved to near-control values by 84 days. Significant periosteal-to bone contact did not appear until 45 days postoperatively. CONCLUSION: Our results promote the use of methods of brow fixation that support mobilized soft tissues for a minimum of 6 weeks, until the elevated periosteum has significantly readhered to the underlying bone. PMID- 12533143 TI - Evaluation of pericranial skull adherence during healing in the rabbit model. AB - BACKGROUND: The endoscopic brow-lift is a popular technique for rejuvenation of the aging brow and forehead. Long-lasting results depend on readherence of the pericranium to the underlying skull in the newly elevated position. Determination of the time required for pericranial readherence to occur is important when considering optimal brow fixation time postoperatively; however, few studies of pericranial healing exist in the literature. OBJECTIVE: To quantify the time required for pericranial adherence after pericranial elevation in a rabbit model. DESIGN: Anesthetized New Zealand white rabbits underwent elevation of a pericranial flap on day 0. The flap was then repositioned and the skin sutured. One unoperated-on group served as a control. A tensiometer was used to measure the force required to separate the pericranial flap from the skull of the control animals and of test animals killed on postoperative days 3, 5, 8, 10, 13, 17, 20, 25, and 28. Statistical analysis was performed to determine the effect of healing time on the strength of pericranial readherence. RESULTS: There was a statistically significant decrease in the force required for pericranial separation at 3 and 5 days after surgery compared with the control group. By 8 days postoperatively and throughout the subsequent times examined, no statistically significant differences from the control group were observed. CONCLUSION: In this rabbit model, pericranial adherence (as measured by tensile strength) is decreased postoperatively and does not return to baseline levels until postoperative day 8. PMID- 12533144 TI - Testing a device to replace the leech for treating venous congestion. AB - OBJECTIVE: To test the effectiveness of a device designed to promote decongestion and tissue survival of a fasciocutaneous flap during 15 hours of complete venous obstruction. METHODS: In a porcine model, a 9 x 7-cm fasciocutaneous flap was elevated and the associated veins were clamped, causing complete venous obstruction for 15 hours in 6 control and 6 treatment animals. Up to 3 devices were used to treat the flap in a predetermined pattern. Control flaps were not treated. Measures of treatment efficacy included blood volumes removed; changes in skin color, surface perfusion, and tissue oxygen tension; and end point histologic findings. RESULTS: Control flaps were characterized by progressive darkening of skin color, undetectable surface perfusion, and low levels of oxygen tension. Histologic assessment showed severe congestion and extravasation of blood and distinct signs of necrosis. In contrast, treated flaps had significant improvements in skin color, surface perfusion, and subcutaneous oxygen tension. Histologic analysis showed little, if any, congestion and no signs of necrosis. Mean blood volume removed was 29.5 mL/h. CONCLUSION: The device was effective in decongesting a large area of tissue during 15 hours of complete outflow obstruction, based on quantitative measurements of tissue health and viability. PMID- 12533145 TI - Expression of endothelin 1 in rat random-pattern skin flaps treated with topical nifedipine. AB - OBJECTIVE: To evaluate the relative tissue concentrations of the endogenous vasoactive peptide endothelin 1 (ET-1) in random-pattern skin flaps (RPSF) treated with either topical anti-ischemic drug therapy (nifedipine) or placebo. DESIGN: Prospective, randomized, placebo-controlled therapeutic trial. SUBJECTS: Adult male Sprague-Dawley rats. INTERVENTION: Experimental subjects underwent caudally based RPSFs using the modified McFarlane technique. Subjects received either topical anti-ischemic drug therapy (nifedipine; n = 6) or inert carrier ointment (placebo; n = 6). Treatment was initiated immediately following flap closure and continued every 6 hours for 5 days. At the end of the treatment period, the animals were killed and the concentration of ET-1 was determined using enzyme-linked immunosorbent assay. Representative tissues from nifedipine- and placebo-treated skin flaps were also analyzed for ET-1 using immunohistochemical stains. RESULTS: The ET-1 levels in the distal (necrotic) flap segments were increased by 4.53 pg/mL over baseline (nonnecrotic) flaps in the placebo-treated animals and decreased by 4.70 pg/mL below baseline in the nifedipine-treated group (P =.03). CONCLUSIONS: The correlation between tissue levels of ET-1 and the severity of tissue necrosis suggests that ET-1 may play a pivotal role in ischemic injury of RPSFs. Moreover, treatment with topical nifedipine may antagonize the vasoconstrictive effects of ET-1. Although immunohistochemical analysis revealed ET-1 staining within the flap microvasculature, no quantitative differences were detected between the nifedipine- and placebo-treated flaps. Further studies are needed to define the role of ET-1 in RPSF necrosis. PMID- 12533146 TI - The effect of tension on patency of rat femoral artery anastomoses. AB - OBJECTIVE: To determine experimentally the role of tension in the failure of microarterial anastomoses. METHODS: Sixteen microarterial anastomoses were performed in femoral arteries from adult Sprague-Dawley male rats after resecting 0, 1, 2, or 3 mm. The percentage of vessel excised was calculated. The vessel was then examined in approximately 1 week for patency. Next, 5 femoral arteries were excised and length-tension curves generated. RESULTS: All vessels were patent in which less than 20% of the length was resected. Clinically, only vessels under extreme tension failed. Length-tension curves suggested 2 zones of vessel extensibility with a breakpoint between them. Extension of the vessel beyond this breakpoint required extreme tension. CONCLUSIONS: Moderate tension at a microarterial anastomosis does not lead to anastomotic failure. Failure occurs after the elastic reserve of the vessel is exceeded. PMID- 12533147 TI - Preoperative percutaneous cranial nerve mapping in head and neck surgery. AB - OBJECTIVE: To identify and map the course of the peripheral branches of the cranial nerve preoperatively and percutaneously. DESIGN: Prospective study. Preoperative percutaneous nerve mapping performed prior to the operation under deep sedation or general anesthesia without muscle paralysis. SETTING: Private office surgery suite, freestanding surgery center, and regional medical centers. PATIENTS: A total of 142 patients undergoing head and neck surgery and facial plastic surgery between August 1994 and July 1999. INTERVENTION: Monopolar probe was used for nerve stimulation. Electromyographic reading was done through intramuscular bipolar recording electrodes. The equipment used was a nerve monitor. RESULTS: The mandibular divisions were tested in 142 cases, the frontal division in 60 cases, the accessory nerve in 12 cases, and the hypoglossal nerve in 3 cases. Satisfactory mappings were obtained in 115 cases of the mandibular division, 49 cases of the frontal division, 8 cases of the accessory division, and 1 case of the hypoglossal nerve. CONCLUSIONS: Preoperative percutaneous nerve mapping is a new method of identifying the location of the peripheral branches of the cranial nerves. Identifying and mapping the course of peripheral branches of the cranial nerves safely assists the head and neck surgeon in the placement of incisions in a favorable location and in the dissection of the area involving the nerves. Mapping alerts the surgeon to an area containing a nerve and allows the surgeon to avoid just the specific area where a nerve is present, preventing large-scale abandonment of unmapped areas for fear of potential nerve damage. PMID- 12533148 TI - Optimal design of O-to-Z flaps for closure of facial skin defects. AB - OBJECTIVE: To determine the optimal design of an O-to-Z flap for closure of facial skin defects. METHODS: Prospective cadaver study. Multiple 2-cm-diameter circular skin defects were created in fresh cadavers. Three types of O-to-Z flaps were designed, varying the angle of a curved line about concentric radii of the defect: acute, intermediate, or wide angle flap. The tension of closure of each was measured and compared at different lengths of incision and extents of undermining. RESULTS: The acute angle flap had a significantly lower closing tension at all lengths of incision and extents of undermining than the intermediate and wide angle flaps. Increasing the amount of undermining alone without incising the flap did not significantly decrease the closing tension. Incising the acute angle flap to 4 radii created a nearly tension-free closure. CONCLUSIONS: The optimal design for reducing tension of the O-to Z flap is an acute angle flap. The optimal length of incision and undermining necessary to minimize closing tension is discussed. PMID- 12533149 TI - Effects of transforming growth factor Beta and insulinlike growth factor 1 on the biomechanical and histologic properties of tissue-engineered cartilage. AB - OBJECTIVE: To investigate the histologic and biomechanical properties of rabbit tissue-engineered cartilage exposed to insulinlike growth factor 1 and transforming growth factor beta. DESIGN: Controlled study. SUBJECTS: New Zealand white rabbits aged 3 to 4 weeks. INTERVENTION: A mean of 3.42 million rabbit chondrocytes were placed onto 2 x 1-cm polyglycolic/poly-L-lactic acid mesh templates. One group (n = 21) was placed in complete medium for 4 days. The experimental group (n = 19) was placed into complete medium with insulinlike growth factor 1 (50 ng/mL) and transforming growth factor beta (1 ng/mL). After 96 hours the templates were removed and implanted into the dorsum of the donor rabbit. The templates were harvested after 8 weeks and subjected to gross, histologic, and biomechanical testing. RESULTS: All samples showed histologic characteristics consistent with normal cartilage. No statistically significant differences were found with biomechanical testing between the control and experimental groups. CONCLUSION: In spite of more promising results from earlier studies, these results do not support improved histologic features or mechanical performance with the addition of insulinlike growth factor 1 and transforming growth factor beta to the chondrocyte/template complex. PMID- 12533150 TI - Effects of growth factors on the quality of ex vivo-generated cartilage. PMID- 12533151 TI - Defect repair in the rat mandible with bone morphogenic proteins and marrow cells. AB - OBJECTIVE: To investigate the ability of a bone growth factor mixture and bone marrow cells to repair a critical size defect of the rat mandibular body. DESIGN: Prospective, randomized controlled trial. SUBJECTS: Thirty-seven male Fischer rats. INTERVENTIONS: Critical size defects 4 mm in diameter were created in the left mandibular bodies of the rats. The defects were filled with a bone marrow cell suspension (group 1), a synthetic bone matrix consisting of bovine collagen and calcium hydroxyapatite cement (group 2), the matrix and marrow cells (group 3), the matrix with 100 micro g of bone growth factor mixture (group 4), or the matrix with bone growth factor mixture and marrow cells (group 5). Animals were killed after 8 weeks, and the nondemineralized specimens were processed histologically. Specimens from group 1 were not processed because there was no grossly appreciable bone regeneration. Stereologic techniques were used to determine and compare the volume fractions and volume estimates of mature bone, new bone, osteoid, marrow, remaining cement, and fibrous tissue in each defect. RESULTS: Volumes of mature bone, new bone, and remaining cement did not differ significantly among the groups (P =.30 for mature bone, P =.17 for new bone, and P =.34 for cement). However, group 4 and 5 specimens contained significantly more osteoid and larger marrow spaces than did the group 2 and 3 specimens (P<.001 for both). The specimens in groups 2 and 3 contained significantly more fibrous tissue ingrowth than did those in groups 4 and 5 (P<.001). CONCLUSION: The synthetic bone substitute containing bone growth factor mixture was effective in stimulating new bone and osteoid development in the rat mandibular model. PMID- 12533152 TI - Use of AlloDerm implant to prevent frey syndrome after parotidectomy. AB - OBJECTIVE: To evaluate the effectiveness of AlloDerm, an acellular human dermal matrix graft, as an interpositional physical barrier to prevent the development of Frey syndrome after parotidectomy. METHODS: The 30 patients included in the study were divided into 3 groups of 10. In group 1 (study group), patients underwent superficial parotidectomy with placement of an AlloDerm graft (LifeCell Corp, Branchburg, NJ). In group 2 (control) patients had superficial parotidectomy without placement of an interpositional barrier. In group 3 (control), patients underwent deep-plane rhytidectomy without disruption of the parotid fascia. All were evaluated after 1 year and questioned about gustatory sweating. Subjective assessment of Frey syndrome was documented when patients experienced gustatory sweating, even if they were not perturbed by the symptom. The Minor starch-iodine test was performed in each patient for objective assessment. RESULTS: The incidence of subjective Frey syndrome was observed in 1 patient in group 1 and 5 patients in group 2. The incidence of objective Frey syndrome was noted in 2 patients in group 1 and 8 patients in group 2. Both subjective and objective differences in incidence of Frey syndrome were statistically significant. None of the group 3 patients had subjective or objective Frey syndrome. Two patients in group 1 and 3 patients in group 2 developed a transient seroma or sialocele that resolved with conservative management. CONCLUSION: The use of AlloDerm graft as an interpositional barrier improves parotidectomy outcome by reducing the incidence of Frey syndrome. PMID- 12533153 TI - Effect of air bags and restraining devices on the pattern of facial fractures in motor vehicle crashes. AB - OBJECTIVE: To examine the relationship between the use of restraining devices and the incidence of specific facial fractures in motor vehicle crashes. DESIGN: Retrospective analysis of patients with facial fractures following a motor vehicle crash. SETTING: University of Alabama at Birmingham Hospital level I trauma center from 1996 to 2000. PATIENTS: Of 3731 patients involved in motor vehicle crashes, a total of 497 patients were found to have facial fractures as determined by International Classification of Diseases, Ninth Revision (ICD-9) codes. Facial fractures were categorized as mandibular, orbital, zygomaticomaxillary complex (ZMC), and nasal. RESULTS: Use of seat belts alone was more effective in decreasing the chance of facial fractures in this population (from 17% to 8%) compared with the use of air bags alone (17% to 11%). The use of seat belts and air bags together decreased the incidence of facial fractures from 17% to 5%. CONCLUSIONS: Use of restraining devices in vehicles significantly reduces the chance of incurring facial fractures in a severe motor vehicle crash. However, use of air bags and seat belts does not change the pattern of facial fractures greatly except for ZMC fractures. Air bags are least effective in preventing ZMC fractures. Improving the mechanics of restraining devices might be needed to minimize facial fractures. PMID- 12533154 TI - Letter to the editor: modified mohs technique for Basal cell carcinomas of the head and neck. PMID- 12533155 TI - Hair transplantation in women: treating female pattern baldness and repairing distortion and scarring from prior cosmetic surgery. AB - The role of hair transplantation in men is well established. In women, the procedure is much less common, but has a definite role in the management of female pattern baldness and the repair of alopecic scarring and hairline distortion as a result of prior facial plastic surgery. When performing hair transplantation in women, there are differences in technique from that used in men to consistently achieve excellent results and minimize complications. Over the past 3 years, I have performed 86 hair transplant procedures on women. Most of these cases were for female pattern baldness. The techniques used and typical results are presented herein. When performed properly for the appropriate indications, hair transplantation is an effective procedure with a very high level of patient satisfaction. PMID- 12533157 TI - Foreign body granulomas caused by polymethylmethacrylate microspheres: successful treatment with allopurinol. PMID- 12533159 TI - Some words that matter. PMID- 12533160 TI - Treatment of toxic epidermal necrolysis with high-dose intravenous immunoglobulins: multicenter retrospective analysis of 48 consecutive cases. AB - OBJECTIVE: To evaluate the effect of high-dose intravenous immunoglobulin (IVIG) in toxic epidermal necrolysis (TEN), parameters that may affect response to treatment, and the effect of different IVIG batches on Fas-mediated cell death. DESIGN: Multicenter retrospective analysis of 48 consecutive TEN patients treated with IVIG. SETTING: Fourteen university hospital dermatology centers in Europe and the United States. PATIENTS: Forty-eight patients with TEN (skin detachment >10% of their body surface [mean, 44.8%; range, 10%-95%]). INTERVENTIONS: Infusion of IVIG in all patients (range, 0.8-5.8 g/kg), and analysis of the ability of different IVIG batches to inhibit Fas-mediated cell death. MAIN OUTCOME MEASURES: Objective response to IVIG treatment, final outcome at day 45, parameters that may affect response to IVIG treatment, and tolerance. RESULTS: Infusion of IVIG (mean total dose, 2.7 g/kg [range, 0.65-5.8 g/kg]; mean consecutive days, 4 [range, 1-5 days]) was associated with a rapid cessation (mean, 2.3 days [range, 1-6 days]) of skin and mucosal detachment in 43 patients (90%) and survival in 42 (88%). Patients who responded to IVIG had received treatment earlier in the course of disease and, on average, higher doses of IVIG. Furthermore, analysis of 35 IVIG batches revealed significant batch-to-batch variations in the capacity of IVIG to inhibit Fas-mediated cell death in vitro. CONCLUSIONS: Early infusion of high-dose IVIG is safe, well tolerated, and likely to be effective in improving the survival of patients with TEN. We recommend early treatment with IVIG at a total dose of 3 g/kg over 3 consecutive days (1 g/kg per day for 3 days). PMID- 12533161 TI - Intravenous immunoglobulin treatment for Stevens-Johnson syndrome and toxic epidermal necrolysis: a prospective noncomparative study showing no benefit on mortality or progression. AB - BACKGROUND: It has been proposed that Fas-Fas ligand interaction was responsible for the apoptosis of epidermal cells in Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) and that high doses of intravenous immunoglobulin (IVIG) could help patients by blocking the apoptosis. OBJECTIVE: To study the effects of IVIG on SJS and TEN. DESIGN: Prospective open trial. SETTING: Referral center of a university hospital. PATIENTS: Thirty-four consecutive patients admitted for SJS (n = 9), SJS-TEN (n = 5), or TEN (n = 20) a mean of 4.3 days after onset. INTERVENTION: A dose of 2 g/kg of IVIG was administered within 2 days (half doses or full doses over a longer period for patients with low creatinine clearance). MAIN OUTCOME MEASURES: Detached plus detachable proportions of the total body surface area measured before and after treatment and predicted death rate estimated on admission with a validated prognostic score. RESULTS: Epidermal detachment involved a mean +/- SD 19% +/- 16% of the total body surface area on admission and 32% +/- 26% after IVIG treatment (progression in 22 of 34 cases, including most patients referred early). The prognostic score predicted 8.2 deaths (24%); 11 were observed (32%; 95% confidence interval, 17%-51%). Most deaths occurred in elderly patients who had initially impaired renal function. CONCLUSIONS: The confidence interval of the observed death rate excludes a dramatic decrease in mortality. No measurable effect was observed on the progression of detachment or on the speed of reepidermalization. These results do not support the routine use of IVIG treatment for patients with SJS or TEN, especially in cases of impaired renal function. PMID- 12533162 TI - Analysis of intravenous immunoglobulin for the treatment of toxic epidermal necrolysis using SCORTEN: The University of Miami Experience. AB - BACKGROUND: Toxic epidermal necrolysis (TEN) is a rare, life-threatening condition caused by certain medications. Keratinocytes affected by TEN have been found to undergo apoptosis mediated by Fas-FasL interactions. Treatment with intravenous immunoglobulin (IVIG) has been proposed to inhibit this interaction. OBJECTIVE: To demonstrate the effectiveness of IVIG therapy in reducing mortality in patients with TEN. DESIGN: A retrospective analysis of 16 consecutive patients with TEN who were treated with IVIG. The SCORTEN system, a validated predictor of TEN mortality, was used to analyze the data of these patients. Using SCORTEN, we compared the predicted mortality of our patient population with observed mortality. SETTING: Dermatology inpatient unit at a university-affiliated hospital. INTERVENTION: All 16 patients received IVIG treatment daily for 4 days. Fifteen patients received 1 g/kg per day and 1 patient received 0.4 g/kg per day. MAIN OUTCOME MEASURES: For each patient, causes of TEN and other medical problems were documented prior to IVIG therapy, as were the 7 independent SCORTEN risk factors. RESULTS: One patient died. Based on the SCORTEN system, 5.81 patients were expected to die. These mortality rates were compared using the standardized mortality ratio (SMR) analysis ([Sigma observed deaths/Sigma expected deaths] x 100) to determine the efficacy of this treatment, which showed that patients with TEN treated with IVIG were 83% less likely to die than those not treated with IVIG (SMR = 0.17; 95% confidence interval, 0.0-0.96). CONCLUSION: Based on comparison of our observed mortality rate with the SCORTEN-predicted mortality rate, treatment with IVIG significantly decreased mortality in patients with TEN. PMID- 12533163 TI - Drug-induced, Ro/SSA-positive cutaneous lupus erythematosus. AB - OBJECTIVE: To study the clinical and immunopathologic findings of drug-induced, Ro/SSA-positive cutaneous lupus erythematosus (CLE). DESIGN: Retrospective medical and laboratory record review. SETTING: Immunodermatology Division of Johns Hopkins Hospital (Baltimore, Md). PATIENTS: Of 120 patients found to have anti-Ro/SSA antibodies by hemagglutination and/or double immunodiffusion, 70 had clinical and immunopathologic confirmation of CLE. Fifteen of these 70 patients had a history of new drug exposure, defined as less than 6 months, associated with disease development. RESULTS: The disease-associated drugs included hydrochlorothiazide (5 patients), angiotensin-converting enzyme inhibitors (3 patients), calcium channel blockers (3 patients), interferons (2 patients), and statins (2 patients). The most common presentations were photodistributed diffuse erythema and subacute CLE-type lesions without evidence of significant systemic disease. All specimens revealed interface dermatitis and fine granular IgG deposition along the basement membrane zone and throughout the epidermis. Most patients experienced improvement or resolution of clinical lesions within 8 weeks and decrease of Ro/SSA titers within 8 months after discontinuation of drug treatment. CONCLUSIONS: Antihypertensive drugs are the most commonly associated with Ro-positive CLE. Clinical and immunopathologic features of this drug-induced variant do not seem to differ from the idiopathic disease. In most cases, the disease improves or resolves on discontinuation of the offending drug treatment. It is not known if these drugs precipitate disease in patients who have subclinical disease. Drug-induced Ro/SSA-positive CLE should be included on the differential diagnosis in patients presenting with photosensitive or subacute CLE type eruptions. PMID- 12533164 TI - Low-dose thalidomide therapy for refractory cutaneous lesions of lupus erythematosus. AB - BACKGROUND: Thalidomide is an anti-inflammatory agent and an immunomodulator that inhibits the production of tumor necrosis factor alpha. It has shown promise as a treatment option for the cutaneous manifestations of lupus erythematosus (LE). OBJECTIVE: To assess the degree of clinical response per subtype of cutaneous lupus, the duration of therapy before documented clinical improvement, and the incidence of adverse effects, including peripheral neuropathy, with low-dose thalidomide therapy at 100 mg daily in the treatment of refractory cutaneous lesions of LE. METHODS: This retrospective medical record review of patients with refractory cutaneous manifestations of LE is one of the largest modern series in the literature. There were 29 patients seen at the Department of Dermatology, Wake Forest University School of Medicine (Winston-Salem, NC), who were unresponsive to conventional agents including antimalarial agents, and who started treatment between 1998 and 2000. Twenty-three patients who took the drug for 1 month or more were included in the analysis. Clinical responses were assessed by the investigators based on statements of improvement listed in the clinic notes and were categorized as "no response," "partial response," and "complete response." Partial response was classified as either 75% or greater or less than 75% improvement. The incidence of adverse effects including peripheral neuropathy was determined. RESULTS: Of the 23 patients, 17 (74%) demonstrated complete resolution of the cutaneous manifestations of LE, whereas 3 patients (13%) demonstrated 75% or greater partial improvement; 3 patients (13%) had less than 75% partial clinical improvement; and 21 patients (91%) who demonstrated a complete or partial response did so within 8 weeks of initiating thalidomide therapy. CONCLUSIONS: Based on the results of this case series, we believe that thalidomide should be given prime consideration as a treatment for antimalarial drug-resistant interface lesions of LE. The design of prospective, randomized, double-blind, placebo-controlled trials for this indication is warranted. PMID- 12533165 TI - Amelioration of body odor after intracutaneous axillary injection of botulinum toxin A. AB - BACKGROUND: Body odor is a ubiquitous phenomenon. It is commonly attributed to sweating and noted explicitly in the axillary area. Botulinum toxin A has recently been shown to be effective for axillary hyperhidrosis. Its effect on axillary odor, however, is unknown. OBSERVATIONS: Sixteen healthy volunteers were injected with botulinum toxin A (Dysport, 100 U dissolved in 0.9% sodium chloride solution) in one axilla and 0.9% sodium chloride solution in the other axilla in a randomized, double-blinded fashion. After 7 days, body odor was assessed by a T shirt sniff test. A significant reduction of odor intensity was observed for the botulinum toxin A-treated side. The smell was also rated significantly less unpleasant. CONCLUSIONS: These findings suggest that botulinum toxin A can ameliorate or even improve body odor. The underlying mechanisms may include interference with skin microbes and denervation of apoeccrine sweat glands, but this remains to be further investigated. PMID- 12533166 TI - Ultraviolet radiation in alpine skiing: magnitude of exposure and importance of regular protection. AB - BACKGROUND: Participation in outdoor alpine sports has been increasing over the last decade. Ultraviolet exposure levels for these activities can be extreme owing to the venue altitudes. The purpose of this study was to measure the levels of UV-A and UV-B radiation exposure incurred by performance skiers in a typical alpine venue and to determine the need for protection in that environment. OBSERVATIONS: Total UV-B exposure ranged from 12 to 185 mJ/cm(2) (0.5-7.6 times the minimum erythemal dose [MED] for individuals with type II skin). Ten percent of subjects received more than 1 MED/h at peak exposure times. Daily UV-A exposure ranged from 10.6 to 28 J/cm( 2) (daily average, 0.55 minimum melanogenic dose). CONCLUSIONS: Alpine skiers with skin types I and II are exposed to measurable erythemal and suberythemal levels of UV radiation repeatedly over the long term at levels that can cause photodamage to their skin and potentially increase their later risk for skin cancer. Programs should be developed to educate skiers regarding the need for UV protection. Given the high levels of exposure noted, skiers should use UV protective measures, including sun protection factor 30 broad-spectrum sunscreen. PMID- 12533167 TI - Chicago Dermatological Society: the first 100 years. PMID- 12533168 TI - The actinic (solar) keratosis: a 21st-century perspective. PMID- 12533169 TI - Dermatologic history of the ruby laser: the long story of short pulses. PMID- 12533170 TI - Genetic disorders of skin: a decade of progress. PMID- 12533171 TI - Cutaneous reactions to chemotherapy drugs: the art of consultation. PMID- 12533172 TI - Of mice and men: the road to understanding the complex nature of adipose tissue and lipoatrophy. PMID- 12533173 TI - Treatment of toxic epidermal necrolysis: the uncertainty persists but the fog is dispersing. PMID- 12533174 TI - How frequently are drugs associated with the development or exacerbation of subacute cutaneous lupus? PMID- 12533175 TI - Nodules on the arm of a diabetic patient. PMID- 12533176 TI - Asymptomatic pedunculated nodule on the leg of a 75-year-old man. PMID- 12533177 TI - Two dark plaques on the thigh. PMID- 12533178 TI - Sudden onset of multiple painful erythematous papules and pustules. PMID- 12533179 TI - Sentinel lymph node biopsy and adjuvant therapy for melanoma: evidence revisited. PMID- 12533181 TI - Reticular telangiectatic erythema associated with an implantable cardioverter defibrillator: an underpublished entity? PMID- 12533182 TI - Newer neutrophilic dermatoses. PMID- 12533183 TI - Neutrophilic dermatosis of the dorsal hands: a variant of erythema elevatum diutinum? PMID- 12533184 TI - Visibility of neurofibromatosis 1 and psychiatric morbidity. PMID- 12533185 TI - Cutaneous manifestation of reactivation of Chagas disease in a renal transplant patient: long-term follow-up. PMID- 12533186 TI - Hair changes in women from the Women's Interagency HIV Study. PMID- 12533191 TI - Cross-linking of cellular proteins by tissue transglutaminase during necrotic cell death: a mechanism for maintaining tissue integrity. AB - Tissue transglutaminase (tTG) is a Ca(2+)-dependent enzyme which cross-links proteins via epsilon(gamma-glutamyl)lysine bridges. There is increasing evidence that tTG is involved in wound repair and tissue stabilization, as well as in physiological mechanisms leading to cell death. To investigate the role of this enzyme in tissue wounding leading to loss of Ca(2+) homoeostasis, we initially used a model involving electroporation to reproduce cell wounding under controlled conditions. Two cell models were used whereby tTG expression is regulated either by antisense silencing in ECV 304 cells or by using transfected Swiss 3T3 cells in which tTG expression is under the control of the tet regulatory system. Using these cells, loss of Ca(2+) homoeostasis following electroporation led to a tTG-dependent formation of highly cross-linked proteinaceous shells from intracellular proteins. Formation of these structures is dependent on elevated intracellular Ca(2+), but it is independent of intracellular proteases and is near maximal after only 20 min post-wounding. Using labelled primary amines as an indicator of tTG activity within these 'wounded cells', we demonstrate that tTG modifies a wide range of proteins that are present in both the perinuclear and intranuclear spaces. The demonstration of entrapped DNA within these shell structures, which showed limited fragmentation, provides evidence that the high degree of transglutaminase cross-linking results in the prevention of DNA release, which may serve to dampen any subsequent inflammatory response. Comparable observations were shown when monolayers of cells were mechanically wounded by scratching. In this second model of cell wounding, redistribution of tTG activity to the extracellular matrix was also demonstrated, an effect which may serve to stabilize tissues post-trauma, and thus contribute to the maintenance of tissue integrity. PMID- 12533193 TI - Bifunctional constructs of aspirin and ibuprofen (non-steroidal anti-inflammatory drugs; NSAIDs) that express antibacterial and alkylation activities. AB - Ibuprofen and aspirin are two common non-steroidal anti-inflammatory drugs (NSAIDs). Both NSAIDs have a carbonyl carbon [-C(O)-], which was utilized to attach a nitrogen mustard (N-mustard) ester group or a tripeptide group. The tripeptide consisted of a L-Gly-D-Ala-D-Ala sequence, where D-Ala-D-Ala is the reactive site for antibacterial activity and L-Gly serves as a linker to the NSAID carrier drug. The aspirin tripeptide and N-mustard show significant antibacterial activity at >or=5.0 x 10(-5) M against penicillin-susceptible or resistant Escherichia coli. The partition coefficients (log Kow)log P of aspirin and ibuprofen tripeptide drugs were -1.05 and 2.23, respectively. The NSAIDs served as carrier drugs of the N-mustard group which expressed alkylation activity directed towards the nucleophilic primary amine of p -chloroaniline. Hydrolysis of the N-mustard agents yielded the parent structure of aspirin (or ibuprofen) and an N-mustard moiety, 2-[bis(2-chloroethyl)amino]ethanol. The (log Kow)log P for the N-mustard structures of aspirin and ibuprofen were 2.61 and 5.63, respectively. The (log Kow)log P value of 2-[bis(2 chloroethyl)amino]ethanol was 0.56. Fluorescamine was utilized to determine unreacted p -chloroaniline at known time intervals, which permitted calculation of rate constants and rate equations. The aspirin N-mustard agent expressed strong antibacterial activity against a penicillin-resistant bacteria and first order alkylation kinetics. The ibuprofen N-mustard and 2-[bis(2 chloroethyl)amino]ethanol followed second-order alkylation kinetics. All N mustard and tripeptide compounds showed zero violations of the Rule of 5. Values of TPSA (molecular polar surface area), C log P and molecular dipoles were calculated. PMID- 12533192 TI - Functional characterization of the non-catalytic ectodomains of the nucleotide pyrophosphatase/phosphodiesterase NPP1. AB - The ubiquitous nucleotide pyrophosphatases/phosphodiesterases NPP1-3 consist of a short intracellular N-terminal domain, a single transmembrane domain and a large extracellular part, comprising two somatomedin-B-like domains, a catalytic domain and a poorly defined C-terminal domain. We show here that the C-terminal domain of NPP1-3 is structurally related to a family of DNA/RNA non-specific endonucleases. However, none of the residues that are essential for catalysis by the endonucleases are conserved in NPP1-NPP3, suggesting that the nuclease-like domain of NPP1-3 does not represent a second catalytic domain. Truncation analysis revealed that the nuclease-like domain of NPP1 is required for protein stability, for the targeting of NPP1 to the plasma membrane and for the expression of catalytic activity. We also demonstrate that 16 conserved cysteines in the somatomedin-B-like domains of NPP1, in concert with two flanking cysteines, mediate the dimerization of NPP1. The K173Q polymorphism of NPP1, which maps to the second somatomedin-B-like domain and has been associated with the aetiology of insulin resistance, did not affect the dimerization or catalytic activity of NPP1, and did not endow NPP1 with an affinity for the insulin receptor. Our data suggest that the non-catalytic ectodomains contribute to the subunit structure, stability and function of NPP1-3. PMID- 12533194 TI - Converting systematic reviews to Cochrane format: a cross-sectional survey of Australian authors of systematic reviews. AB - BACKGROUND: Despite the growing reputation and subject coverage of the Cochrane Database of Systematic Reviews, many systematic reviews continue to be published solely in paper-based health care journals. This study was designed to determine why authors choose to publish their systematic reviews outside of the Cochrane Collaboration and if they might be interested in converting their reviews to Cochrane format for publication in the Cochrane Database of Systematic Reviews. METHODS: Cross-sectional survey of Australian primary authors of systematic reviews not published on the Cochrane Database of Systematic Reviews identified from the Database of Abstracts of Reviews of Effectiveness. RESULTS: We identified 88 systematic reviews from the Database of Abstracts of Reviews of Effectiveness with an Australian as the primary author. We surveyed 52 authors for whom valid contact information was available. The response rate was 88 per cent (46/52). Ten authors replied without completing the survey, leaving 36 valid surveys for analysis. The most frequently cited reasons for not undertaking a Cochrane review were: lack of time (78%), the need to undergo specific Cochrane training (46%), unwillingness to update reviews (36%), difficulties with the Cochrane process (26%) and the review topic already registered with the Cochrane Collaboration (21%). (Percentages based on completed responses to individual questions.) Nearly half the respondents would consider converting their review to Cochrane format. Dedicated time emerged as the most important factor in facilitating the potential conversion process. Other factors included navigating the Cochrane system, assistance with updating and financial support. Eighty-six per cent were willing to have their review converted to Cochrane format by another author. CONCLUSION: Time required to complete a Cochrane review and the need for specific training are the primary reasons why some authors publish systematic reviews outside of the Cochrane Collaboration. Encouragingly, almost half of the authors would consider converting their review to Cochrane format. Based on the current number of reviews in the Database of Abstracts of Reviews of Effectiveness, this could result in more than 700 additional Cochrane reviews. Ways of supporting these authors and how to provide dedicated time to convert systematic reviews needs further consideration. PMID- 12533195 TI - Poorly differentiated carcinoma arising from adenolymphoma of the parotid gland. AB - BACKGROUND: There is only one previous case report of a poorly differentiated carcinoma arising from an adenolymphoma of the parotid gland (Warthin's tumour). The absence of clinical symptoms, and the aspecificity of the radiological pattern make the diagnosis very difficult. CASE PRESENTATION: We here report the case of a 73-year-old man with Warthin's tumour who was brought to our attention because of a swelling in the parotid region. CONCLUSIONS: In this case with an atypical clinical presentation, the intra-operative examination of a frozen section of the parotid mass allowed us to diagnose the malignant tumour correctly and consequently undertake its radical excision. PMID- 12533197 TI - Proceedings of a symposium in honor of Professor Sir John Meurig Thomas. PMID- 12533198 TI - Synthesis and microstructural analysis of Si3N4 nanorods. AB - Alpha-Si3N4 nanorods with 20-80 nm width were synthesized by carbothermal reduction of SiO with amorphous activated carbon (AAC) as a reductant. Microstructural characterization of the synthesized nanorods was carried out by high resolution transmission electron microscopy (HRTEM) and energy dispersive X ray analysis. Many Si3N4 nanorods were found to be twisted. Each twisted nanorod contained several straight Si3N4 parts. The straight parts had the rod axes orientated along the (1010) direction, which is the closest packing direction of alpha-Si3N4. There were two kinds of joints between the two adjacent straight Si3N4 parts. Formation mechanism of the Si3N4 nanorods is discussed. PMID- 12533199 TI - Strain analysis of Si by FEM and energy-filtering CBED. AB - Lattice strains around a platelet oxygen precipitate in Si wafer is studied by energy filtering convergent-beam electron diffraction (CBED) and calculations based on the finite element method (FEM). Local lattice strains are measured from CBD patterns obtained with a probe size less than 2 nm in a specimen thicker than 450 nm. Strains measured are compressive along a direction normal to a plate of the precipitation and tensile along a direction parallel to the plate. Two dimensional stress fields near the precipitate are obtained with FEM computer analyses by fitting the measured strains. It appears that shear stresses are concentrated at the end of the precipitate edge and the maximum shear stress at an interface between the precipitate and the Si-matrix is 1.9 GPa. It is demonstrated that a combination of the energy filtering CBED and FEM is very useful for the study of local strains near interfaces in semiconductor devices, in particular for the study of stress fields that are too steep for application of the conventional CBED technique. PMID- 12533200 TI - To-and-fro motion of W5Si3 on a beta-Si3N4 substrate at elevated temperatures. AB - Behavior of fine crystalline particles of W5Si3 on a beta-Si3N4 substrate at high temperatures was observed by an in situ heating experiment in a transmission electron microscope. Some of the fine particles of W5Si3 moved in a to-and-fro manner. PMID- 12533201 TI - The nanostructures of amorphous silicas. AB - Topologically modeled amorphized silica structures have been refined using a molecular dynamics simulation technique. Several metastable structures with substantially different medium-range connectivities, as characterized by primitive ring statistics, were obtained. Whereas the total correlation function is insensitive to these differences, the first step diffraction peak derived from energy-filtered electron diffraction shows a promising correlation to medium range structure. PMID- 12533202 TI - Electron microscopy study of novel Pt nanowires synthesized in the spaces of silica mesoporous materials. AB - Structures of Pt-nanowires, synthesized in channels of silica mesoporous materials MCM-41, SBA-15 and MCM-48, were investigated by transmission electron microscopy. One dimensional (1D) Pt-nanowires were formed inside the channels of the MCM-41, and were single crystals with a length of several tens to several hundreds nanometers and a diameter of ca. 3 nm pt-nanowires synthesized in SBA-15 formed a new 3D-network following 3D-pore geometry of SBA-15; that is, the main 1D-channels are interconnected to each other through randomly distributed tunnels. These Pt-nanowires showed a well single crystalline. MCM-48 has two non intersecting chiral channels, and Pt-networks were mostly formed in one of the two channels. Therefore the networks were also chiral; however, the chirality of Pt-networks remained to be determined. It was shown that all Pt-nanowires were formed following the channel geometries of silica mesoporous materials used. PMID- 12533203 TI - Chemistry with the electron microscope: some highlights in a lifetime's journey. AB - The supreme advantages of electron microscopy (EM) in the chemical sciences are briefly recalled: By judicious use of electron optical techniques, vital information of a structural, mechanistic, compositional, and often of an electronic kind may be retrieved. Not only are insights gained (through EM) into the existence of whole new families of structures hitherto unperceived, but one also uncovers the structural characteristics of imperfections in solids. And it is often the case that these imperfections reflect or suggest altogether new structures, hitherto unconceived. EM is , therefore, a powerful agent for aiding chemical synthesis of new materials. This is particularly important in the field of heterogeneous catalysis, since altogether new types of catalytic materials may be, on the one hand, defined, described, identified, and characterised, and, on the other, designed and synthesised. There is also the ever-improving role of the electron microscope as an analytical tool: Very few other techniques within reach of the chemist can rival it in tis sensitivity and detection limits. (Scanning instruments now permit the imaging and the identification of nanoclusters consisting of just a few atoms.) But there are numerous other branches of chemistry besides catalysis and surface science where EM proves invaluable, as we outline herein, in elucidating structure-property or composition and structure interrelationships. PMID- 12533205 TI - Bismuth nanowires for potential applications in nanoscale electronics technology. AB - Nanowires of bismuth with diameters ranging from 10 to 200 nm and lengths of 50 microm have been synthesized by a pressure injection method. Nanostructural and chemical compositional studies using environmental and high resolution transmission electron microscopy with electron stimulated energy dispersive X-ray spectroscopy have revealed essentially single crystal nanowires. The high resolution studies have shown that the nanowires contain amorphous Bi-oxide layers of a few nanometers on the surface. In situ environmental high resolution transmission electron microscopy (environmental-HRTEM) studies at the atomic level, in controlled hydrogen and other reducing gas environments at high temperatures demonstrate that gas reduction can be successfully applied to remove th oxide nanolayers and to maintain the dimensional and structural uniformity of the nanowires, which is key to attaining low electrical contact resistance. PMID- 12533204 TI - Transmission electron microscopy studies of the nanoscale structure and chemistry of Pt50Ru50 electrocatalysts. AB - The structural and chemical heterogeneity of 2.5-nm Pt50Ru50 electrocatalysts was studied by transmission electron microscopy using selected area diffraction, lattice imaging, electron-energy loss spectroscopy, and energy-dispersive X-ray spectroscopy. The catalysts with the highest methanol oxidation activities exhibit oxidation-induced phase separation on the nanoscale to from Pt-rich metal embedded in Ru-rich hydrous and anhydrous oxide. Reduction of the oxide-on metal samples produces a true bimetallic face-centered cubic Pt50Ru50 alloy, with 275 times lower oxidation activity. PMID- 12533206 TI - Synthesis and characterization of quantum dot superlattices. AB - Gold nanoparticles have been synthesized using n-alkylthiol molecules as a passivating agent. By fixing the length of the thiol chain, it is possible to produce nanocrystal arrays, such as 1D chains, 2D arrays of chains and 3D crystals. PMID- 12533207 TI - Tutorial on off-axis electron holography. AB - Through recent years, off-axis electron holography has helped us to understand and to overcome some experimental restrictions in transmission electron microscopy. With development of powerful electron microscopes, slow-scan CCD cameras, and computers, holography is not an academic technique anymore used by specialized laboratories. Holography has proven its wide range of applications in solving real-world problems in materials science and biology. At medium resolution, that is, on nanometer scale, holography allows access to large area phase contrast produced by magnetic fields and electric potentials. In the high resolution domain, holography unveils its power by unscrambling amplitude and phase of the electron wave, resulting in an improved lateral resolution up to the information limit. Holography is a thoroughly quantitative method, and, in combination with the perfect zero-loss filtering inherent to this method, the interpretation of the reconstructed data is strongly simplified. After outlining the basics of holography, in this tutorial we focus on development of a step-by step procedure for recording and reconstruction of holograms. At the end, some recent applications are discussed. PMID- 12533208 TI - Structures of oxide nanobelts and nanowires. AB - We have recently reported the synthesis of one-dimensional nanobelt structures of ZnO, SnO2, In2O3, CdO, Ga2O3, and PbO2 by evaporating the desired commercial metal oxide powders at high temperatures (Science (2001), 291, 1947). The as synthesized oxide nanobelts are pure, structurally uniform, single crystalline, and most of them free from dislocations. The beltlike morphology appears to be a unique and common structural characteristic for the family of semiconducting oxides. In the present article, we focus on the twin and stacking fault planar defects found in oxide nanobelts and nanowires although they are rarely observed. Some interesting and unique growth morphologies are presented to illustrate the roles played by surface energy and kinetics in growth. It is shown that the surfaces of the oxide nanobelts are enclosed by the low-index, low-energy crystallographic facets. The growth morphology is largely dominated by the growth kinetics. PMID- 12533209 TI - Precipitate crystal structure determination in melt spun Mg-1.5wt%Ca-6wt%Zn alloy. AB - A melt-spun Mg-1.5%wtCa-6wt%Zn alloy was analyzed by means of transmission electron microscopy, energy dispersive X-ray spectroscopy, and scanning transmission electron microscopy. The as-solidified alloy exhibited both spherical matrix precipitates and elongated precipitates at the grain boundaries (grain-boundary films). After heat treatment, the alloy showed faceted precipitates (cuboidal shape), mostly on dislocations. It was found that the observed precipitates are the same compound, Ca2Mg6Zn3. As there was no crystallographic data for this compound in the literature, its crystal structure was investigated by comparison of experimental and simulated selected-area electron-diffraction patterns and high-resolution electron microscopy images. This study indicated that Ca2Mg6Zn3 is a trigonal compound with space group P 3 1c and lattice parameters a = 0.97 nm, c = 1.0 nm. PMID- 12533210 TI - Nanostructure manipulation device for transmission electron microscopy: application to titania nanoparticle chain aggregates. AB - Experimental difficulties in studying nanostructures stem from their small size, which limits the use of traditional techniques for measuring their physical properties. We have developed a nanostructure manipulation device to apply tension to chain aggregates mounted in a transmission electron microscope. A 1-mm long slit was cut in the center of a lead-tin alloy disc, measuring 3 mm in diameter and 200 microm in thickness. The disc was heated to about 140 degrees C before it was pressed between two quartz slides. The disc was then thinned by mechanical dimpling and ion milling until holes developed around the slit. The edges of the slit were 0.2 to 3 microm in thickness while the gap between them was up to a few microns. This disc was bonded to the two plates of a cartridge. The slit could be widened or narrowed at controlled speeds of 0.5 to 300 nm/s. The system was tested using titania (TiO2) nanoparticle chain aggregates (NCA) deposited across the slit. The ends of the NCA remained attached to the edges of the slit, which was widened at about 0.7 nm/s. In this way, the NCA was stretched up to 176% of its initial length before breaking. PMID- 12533211 TI - Focused ion beam sectioning and lift-out method for copper and resist vias in organic low-k dielectrics. AB - The focused ion beam lift-out technique for scanning electron microscope (SEM) and transmission electron microscope (TEM) sample preparation was shown to be applicable to copper/low-k dielectric semiconductor technology. High resolution SEM, TEM, and scanning transmission electron microscope analyses were performed on metal contacts and resist vias with no evidence of the interface damage or metal smearing commonly observed with mechanical polishing. Ion milling of the sample ex situ to the substrate provided decoration and adjustment of the exposed plane of the section when necessary for SEM analysis. PMID- 12533212 TI - Implementation of recycling routes for scanning probe microscopy tips. AB - In this work, effective, yet simple, recycling mechanisms for used scanning probe microscopy (SPM) tips were implemented. Comprising a tip profile characterization methodology and specific cleaning procedures, which decontaminate SPM tips whether the contamination nature is known or not, such routes were optimized during numerous tests with brand new, previously used, and already discarded categories of SPM tips. The results show that if the used tip suffered contamination only, but no physical damages, during its scanning lifetime, it becomes readily available for reutilization after the cleansing process, characterizing a recycling route. On the other hand, if the tip went through wear and breakages during its utilization, it still can be decontaminated, but may not be directly reutilized due to its inadequate physical profile. Nevertheless, the methodology developed in this work may yet be applied as part of a more complex recycling route. PMID- 12533214 TI - The pros and cons of apoptosis assays for use in the study of cells, tissues, and organs. AB - Programmed cell death or apoptosis occurs in many tissues during normal development and in the normal homeostasis of adult tissues. Apoptosis also plays a significant role in abnormal development and disease. Increased interest in apoptosis and cell death in general has resulted in the development of new techniques and the revival of old ones. Each assay has its advantages and disadvantages that can render it appropriate and useful for one application, but inappropriate or difficult to use in another. Understanding the strengths and limitations of the assays would allow investigators to select the best methods for their needs. PMID- 12533215 TI - Detection, distribution, and quantification of carbon in steel microstructures by PEELS. AB - This work concerns the use of parallel electron energy loss spectroscopy (PEELS) to investigate the detection, distribution, and quantification of carbon in various steel microstructures generated by rapid cooling rates or by isothermal transformation. The feasibility of detecting C in steels containing very small amounts of carbon was first examined by calculating the minimum detectable mass fraction for a variety of binary Fe-C alloy specimen thicknesses and microscope conditions. These theoretical studies indicated that the detection of carbon in steel microconstituents containing about 0.01 wt.% (or even less) was easily possible with an analytical transmission electron microscope equipped with a LaB6 emitter and a PEEL spectrometer. These theoretical calculations seemed to be reasonable, as it proved possible to make a quantitative PEELS study of the partitioning between the microconstituents ferrite, retained austenite, and martensite found in an ultralow carbon (0.03 wt.%) steel weld metal provided care was taken to avoid hydrocarbon contamination. Studies of both carbon and molybdenum segregation to ferrite/martensite interfaces in an isothermally transformed Fe-C-Mo alloy were also carried out in order to investigate the nature of the "solute drag" effect in this alloy system. PMID- 12533216 TI - Characterization of magnetic nanoparticles using energy-selected transmission electron microscopy. AB - Fe, Co, and Ni magnetic nanoparticles have been characterized using energy selected imaging in a high-resolution transmission electron microscope. The samples comprised Fe/FeO x and Co/CoO x nanoparticles synthesized by inert gas evaporation and a Ni/C nano-composite prepared by a sonochemical method. All of the particles examined were found to be between 5 and 30 nm in size, with the Fe and Co crystals coated in 5-10 nm of metal oxide layer and the Ni metallic crystallites embedded in an amorphous carbon spherical matrix. PMID- 12533217 TI - HRTEM image simulations for the study of ultrathin gate oxides. AB - We have performed high resolution transmission electron microscope (HRTEM) image simulations to qualitatively assess the visibility of various structural defects in ultrathin gate oxides of MOSFET devices, and to quantitatively examine the accuracy of HRTEM in performing gate oxide metrology. Structural models contained crystalline defects embedded in an amorphous 16-A-thick gate oxide. Simulated images were calculated for structures viewed in cross section. Defect visibility was assessed as a function of specimen thickness and defect morphology, composition, size, and orientation. Defect morphologies included asperities lying on the substrate surface, as well as "bridging" defects connecting the substrate to the gate electrode. Measurements of gate oxide thickness extracted from simulated images were compared to actual dimensions in the model structure to assess TEM accuracy for metrology. The effects of specimen tilt, specimen thickness, objective lens defocus, and coefficient of spherical aberration (Cs) on measurement accuracy were explored for nominal 10-A gate oxide thickness. Results from this work suggest that accurate metrology of ultrathin gate oxides (i.e., limited to several percent error) is feasible on a consistent basis only by using a Cs-corrected microscope. However, fundamental limitations remain for characterizing defects in gate oxides using HRTEM, even with the new generation of Cs-corrected microscopes. PMID- 12533218 TI - Measuring thickness changes in thin films due to chemical reaction by monitoring the surface roughness with in situ atomic force microscopy. AB - Measuring the changing thickness of a thin film, without a reference, using an atomic force microscope (AFM) is problematic. Here, we report a method for measuring film thickness based on in situ monitoring of surface roughness of films as their thickness changes. For example, in situ AFM roughness measurements have been performed on alloy film electrodes on rigid substrates as they react with lithium electrochemically. The addition (or removal) of lithium to (or from) the alloy causes the latter to expand (or contract) reversibly in the direction perpendicular to the substrate and, in principle, the change in the overall height of these materials is directly proportional to the change in roughness. If the substrate on which the film is deposited is not perfectly smooth, a correction to the direct proportionality is needed and this is also discussed. PMID- 12533219 TI - Compositional averaging of continuum intensities in multielement compounds. AB - We find no physical basis for traditional mass-fraction weighted averaging of X ray continuum measured in pure elements to estimate continuum production from compounds. Measurements of continuum from pairs of stable isotopes demonstrate that mass alone (the presence of more or fewer neutrons) has no effect on continuum production in electron probe microanalysis. Electron-fraction weighted averaging, which is based on the relative contribution of each constituent element to the electron (or proton) composition of a compound, proved superior to mass-fraction weighted averaging, in predicting the continuum of a compound from the measured continuum of its elements. PMID- 12533220 TI - Symposium Dedicated to Gareth Thomas on the Occasion of his 70th Birthday. PMID- 12533221 TI - Ordering energies and occupancies in doped TiAl, and dedicated ALCHEMI instrumentation. AB - Alloy design has been a lifelong interest of Gareth Thomas, and modern design algorithms include atomistic parameters which are obtainable from new electron microscope techniques such as ALCHEMI. In this paper, we discuss the relevance of ALCHEMI site occupancy measurements to intermetallic alloys, and summarize prior work. The results are found to lie in regions of a site-occupancy diagram (SOC) relating ordering energies to occupancy, as predicted by the Bragg-Williams theory of short-range order. These predictions also explain previous inconsistencies in the ALCHEMI measurements. A diffraction camera and X-ray detector system of novel design is proposed for dedicated ALCHEMI analysis for substitutional and interstitial dopant site-occupancy measurement, and details of the design given. Using this novel hardware design, the data-collection times for two-dimensional ALCHEMI patterns should be reduced by an order of magnitude or more, and the full data collection process automated. The resulting occupancy information can provide essential input parameters for atomistic alloy design algorithms, and can provide entirely new information on interstitial occupancies in minerals, ceramics, semiconductors, and alloys. PMID- 12533222 TI - Electron microscopy observations on the role of twinning in the evolution of microstructures. AB - Twinning plays an important role in phase transformations and can have significant effects on microstructural evolution. Different roles of twinning in the development of microstructures during precipitation and phase transformations are reviewed and illustrated with examples from investigations by high-resolution electron microscopy, including the effect of multiple twinning on the development of Ge precipitates in Al-Ge and Ag-Ge alloys, the twin dissociation of grain boundaries in Au, the formation of hexagonal Si at twin intersections and the effect of twin boundaries on the equilibrium shape of Pb inclusions in Al. PMID- 12533223 TI - Dewetting of liquids on ceramic surfaces at high temperatures. AB - The influence of surface structure and chemistry on high-temperature dewetting of silicate liquids on ceramic surfaces has been investigated. Model systems based on well-defined crystallography and known chemistry have been used to illustrate the effect of surface roughness and chemistry on the dewetting process. Reconstructed ceramic surfaces provide ideal substrates to study effects of surface roughness. It has been shown that the morphology of dewet droplets depend on the length scale and the crystallography of the facets on the surface. Complex pattern formation due to solute redistribution during dewetting is illustrated in the case of SiO2 dewetting on (001) rutile substrates. The role of kinetics on the dewetting process has also been clarified. PMID- 12533224 TI - Structural evolution in amorphous silicon and germanium thin films. AB - The structural evolution in amorphous silicon and germanium thin films has been investigated by high-resolution transmission electron microscopy (HRTEM) in conjunction with autocorrelation function (ACF) analysis. The results established that the structure of as-deposited semiconductor films is of a high density of nanocrystallites embedded in the amorphous matrix. In addition, from ACF analysis, the structure of a-Ge is more ordered than that of a-Si. The density of embedded nanocrystallites in amorphous films was found to diminish with annealing temperature first, then to increase. The conclusions also corroborate well with the results of diminished medium-range order in annealed amorphous films determined previously by a variable coherence microscopy method. PMID- 12533225 TI - Ultrasoft magnetic films investigated with Lorentz tranmission electron microscopy and electron holography. AB - As a tribute to the scientific work of Professor Gareth Thomas in the field of structure-property relationships this paper delineates a new possibility of Lorentz transmission electron microscopy (LTEM) to study the magnetic properties of soft magnetic films. We show that in contrast to the traditional point of view, not only does the direction of the magnetization vector in nano-crystalline films make a correlated small-angle wiggling, but also the magnitude of the magnetization modulus fluctuates. This fluctuation produces a rapid modulation in the LTEM image. A novel analysis of the ripple structure in nano-crystalline Fe Zr-N film corresponds to an amplitude of the transversal component of the magnetization deltaMy of 23 mT and a longitudinal fluctuation of the magnetization of the order of deltaMx = 30 mT. The nano-crystalline (Fe99Zr1)1 xNx films have been prepared by DC magnetron reactive sputtering with a thickness between 50 and 1000 nm. The grain size decreased monotonically with N content from typically 100 nm in the case of N-free films to less than 10 nm for films containing 8 at%. The specimens were examined with a JEOL 2010F 200 kV transmission electron microscope equipped with a post column energy filter (GIF 2000 Gatan Imaging Filter). For holography, the microscope is mounted with a biprism (JEOL biprism with a 0.6 microm diameter platinum wire). PMID- 12533226 TI - In situ TEM studies of metal-carbon reactions. AB - The reactions which occur between amorphous carbon and a number of first transition metals (Ti, Cr, Fe, Co, Ni, and Cu) have been studied by transmission electron microscopy (TEM). The materials are in thin-film form with the metal layer sandwiched between thicker carbon layers. In four cases, the predominant reaction is the graphitization of the amorphous carbon, at temperatures well below 800 degrees C. This is brought about by the elements themselves in the case of Co and Ni, and by metastable carbides in the case of Fe (Fe3C) and Cr (Cr3C2 x). The Ti-C and Cu-C systems do not exhibit graphitization. For the former, only TiC is produced up to 1000 degrees C, while the carbon does not react at all with copper. In situ TEM studies show the mechanism to be of the dissolution precipitation type, which is equivalent to the metal-mediated crystallization process for amorphous silicon and germanium. The heat of graphitization is found to be 18-19 kcal/mol-C by differential scanning calorimetry. PMID- 12533227 TI - Early stages of carbon segregation and precipitation in a sigma = 5 (310)[001] molybdenum tilt boundary. AB - A [Sigma] = 5 (310)[001] tilt grain boundary in molybdenum has been annealed at high temperature in the presence of carbon and observed in high-resolution electron microscopy. The carbon is located at the grain boundary in a 1-nm slab. Two different morphologies coexist. The first one is a grain boundary precipitation while the second one can be considered as a segregation. PMID- 12533228 TI - Nanometric scale investigation of local strain in GaInAs islands by high resolution and analytical TEM. AB - The continuous displacement field within elastically relaxed GaInAs islands was calculated from digitized HREM images of [110] cross sections of In0.35Ga0.65As layers grown on GaAs by molecular beam epitaxy. Experimental maps of the deformations parallel to the interface (epsilonx) and along the growth direction (epsilonz) were drawn and compared with the ones calculated via the finite element method. It was found that epsilonx exp was systematically higher than epsilonx calc and the significant maximum observed for epsilonz exp within the island could not be found for epsilonz calc. These discrepancies were attributed to a variation of the chemical composition in the island. The maps showing the indium concentration gradient drawn from HREM and FE calculations were compared to quantitative profiles for indium concentration obtained by nanometric X-ray microanalysis in TEM. The measured gradient within the island backs our assumption. PMID- 12533229 TI - Physical and magnetic modification of co/pt multilayers by ion irradiation. AB - The microstructure of Co/Pt multilayers with large perpendicular magnetic anisotropy (PMA) was investigated before and after energetic ion irradiation. No pronounced microstructural changes were detected at ion doses sufficient to completely reduce the PMA and cause a spin reorientation transition to in-plane. Ion-induced displacement of Co and Pt atoms near Co/Pt interfaces lead to local "roughening" and Co layer strain relaxation, reducing the PMA. The magnetic domain confinement induced by ion irradiation and magnetic patterning by selective ion irradiation were also investigated. PMID- 12533230 TI - Nanoscale phenomena in synthetic functional oxide heterostructures. AB - This paper reviews nanoscale phenomena such as polarization relaxation dynamics and piezoelectric characterization in model ferroelectric thin films and nanostructures using voltage-modulated scanning force microscopy. Using this technique we show the three-dimensional reconstruction of the polarization vector in lead zirconate titanate (PZT) thin films. Second, the time-dependent relaxation of remanent polarization in epitaxial PZT ferroelectric thin films, containing a uniform two-dimensional grid of 90 degrees domains (c-axis in the plane of the film), has been investigated extensively. The 90 degrees domain walls preferentially nucleate the 180 degrees reverse domains during relaxation. Relaxation occurs through the nucleation and growth of reverse 180 degrees domains, which subsequently coalesce and consume the entire region as a function of relaxation time. In addition we also present results on investigation of the relaxation phenomenon on a very local scale, where pinning and bowing of domain walls has been observed. We also show how this technique is used for obtaining quantitative information on piezoelectric constants and by engineering special structures, and how we realize ultrahigh values of piezoconstants. Last, we also show direct hysteresis measurements on nanoscale capacitors, where there is no observable loss of polarization in capacitors as small as 0.16 microm2 in area. PMID- 12533231 TI - Use of plasmon spectroscopy to evaluate the mechanical properties of materials at the nanoscale. AB - Relationships between volume plasmon excitations and mechanical properties of various materials are considered. Based on systematic evaluation of available data, correlations between the volume plasmon energy, Ep, Young's modulus, Ym, bulk modulus, Bm, shear modulus, Gm, and microhardness, Hm, are established. The resulting correlations indicate that plasmon energies potentially can be used to predict and/or determine the local mechanical properties of technologically important materials, such as metal alloys, semiconductors, and ceramics at the nanometer level. PMID- 12533233 TI - Microvasculature of the olfactory organ in the Japanese monkey (Macaca fuscata fuscata). AB - Olfaction is an important and primitive sense. As its importance has changed with evolution, anatomic adjustments have occurred in its structure and vasculature. Primates are a family of vertebrates that have had to develop their visual system to adapt to the arboreal environment and have evolved from a macrosmatic to a microsmatic species as the optic system has enlarged. This has resulted in anatomic changes of a small but critical area at the base of the brain. This paper describes the three-dimensional vascular anatomy of the olfactory organ of the Japanese monkey (Macaca fuscata fuscata). This is best understood by dividing the organ into three parts: the olfactory tract, olfactory bulb, and olfactory nerves in the nasal mucosa. The bulb can be partitioned into an outer or cortical part and inner or medullary part. The vasculature and tissue were examined grossly and with light microscopy and scanning electron microscopy of vascular corrosion casts. The olfactory tract and bulb were supplied by an arteriole from the anterior cerebral artery on each side. The tract was supplied by capillaries running spirally with a coarse network. At the olfactory bulb, the arteriole ramified into the intracortical and medullary branches that formed capillary networks. The bulbar intracortical capillaries were divided into two layers with different densities and vascular patterns. The capillaries of the superficial layer had a ladder-like pattern. The branches that ran into the medulla of the olfactory bulb were more widely spaced. Twigs from the posterior ethmoidal artery ran along the nerve fiber and formed intra- and extrafascicular networks. Each region of the olfactory organ had characteristic three-dimensional vascular patterns that were related to their cellular architecture. PMID- 12533234 TI - Analysis of the intensity curves obtained by the unidirectional laser oblique illumination technique at different magnifications. AB - The study of the gaussian parameter omega-the approximate width of a peak at half height-of the intensity curves obtained by means of the Unidirectional Laser Oblique Illumination technique employed in the optical microscopy of materials permits introduction of the concept of variation of a surface pattern at different magnifications. This concept establishes how the surface pattern or topography varies among different areas when different magnifications are used. Diffraction gratings and metallic surfaces which have been subjected to different metallographic procedures are analyzed, using objectives of 8x, 16x, and 32x. PMID- 12533235 TI - Phase sensitive demodulation in multiphoton microscopy. AB - Multiphoton laser scanning microscopy offers advantages in depth of penetration into intact samples over other optical sectioning techniques. To achieve these advantages it is necessary to detect the emitted light without spatial filtering. In this nondescanned (nonconfocal) approach, ambient room light can easily contaminate the signal, forcing experiments to be performed in absolute darkness. For multiphoton microscope systems employing mode-locked lasers, signal processing can be used to reduce such problems by taking advantage of the pulsed characteristics of such lasers. Specifically, by recovering fluorescence generated at the mode-locked frequency, interference from stray light and other ambient noise sources can be significantly reduced. This technology can be adapted to existing microscopes by inserting demodulation circuitry between the detector and data collection system. The improvement in signal-to-noise ratio afforded by this approach yields a more robust microscope system and opens the possibility of moving multiphoton microscopy from the research lab to more demanding settings, such as the clinic. PMID- 12533236 TI - Synthesis of electron energy loss spectra for the quantification of detection limits. AB - We describe a method for predicting detection limits of minority elements in electron energy loss spectroscopy (EELS), and its implementation as a software package that gives quantitative predictions for user-specified materials and experimental conditions. The method is based on modeling entire energy loss spectra, including shot noise as well as instrumental noise, and taking into account all the relevant experimental parameters. We describe the steps involved in modeling the entire spectrum, from the zero loss up to inner shell edges, and pay particular attention to the contributions to the pre-edge background. The predicted spectra are used to evaluate the signal-to-noise ratios (SNRs) for inner shell edges from user-specified minority elements. The software also predicts the minimum detectable mass (MDM) and minimum mass fraction (MMF). It can be used to ascertain whether an element present at a particular concentration should be detectable for given experimental conditions, and also to quickly and quantitatively explore ways of optimizing the experimental conditions for a particular EELS analytical task. We demonstrate the usefulness of the software by confirming the recent empirical observation of single atom detection using EELS of phosphorus in thin carbon films, and show the effect on the SNR of varying the acquisition parameters. The case of delta-doped semiconductors is also considered as an important example from materials science where low detection limits and high spatial resolution are essential, and the feasibility of such characterization using EELS is assessed. PMID- 12533237 TI - Chemical microcharacterization of ultrathin iodide conversion layers and adsorbed thiocyanate surface layers on silver halide microcrystals with time-of-flight SIMS. AB - The technique of imaging time-of-flight secondary ion mass spectrometry (TOF SIMS) and dual beam depth profiling has been used to study the composition of the surface of tabular silver halide microcrystals. Analysis of individual microcrystals with a size well below 1 microm from a given emulsion is possible. The method is successfully applied for the characterization of silver halide microcrystals with subpercent global iodide concentrations confined in surface layers with a thickness below 5 nm. The developed TOF-SIMS analytical procedure is explicitly demonstrated for the molecular imaging of adsorbed thiocyanate layers (SCN) at crystal surfaces of individual crystals and for the differentiation of iodide conversion layers synthesized with KI and with AgI micrates (nanocrystals with a size between 10 and 50 nm). It can be concluded that TOF-SIMS as a microanalytical, surface-sensitive technique has some unique properties over other analytical techniques for the study of complex structured surface layers of silver halide microcrystals. This offers valuable information to support the synthesis of future photographic emulsions. PMID- 12533238 TI - Uptake and trafficking of mildly oxidized LDL and acetylated LDL in THP-1 cells does not explain the differences in lysosomal metabolism of these two lipoproteins. AB - Foam cells in the atherosclerotic lesion have substantial cholesterol stores within large, swollen lysosomes. This feature is mimicked by incubating THP-1 macrophages with mildly oxidized low density lipoprotein (LDL). Incubation of THP 1 cells with acetylated LDL produces cytoplasmic cholesteryl ester accumulation rather than lysosomal storage. The differences could be due to differences in uptake and delivery of lipoprotein to lysosomes or to lysosomal and post lysosomal processing events. We compared uptake and lysosomal trafficking of acetylated and oxidized LDL using colloidal gold-labeled lipoproteins. Labeling did not alter cellular cholesterol accumulation. We found that uptake and delivery to lysosomes are not different for acetylated and oxidized LDL. In fact, both oxidized and acetylated LDL can be delivered to the same lysosomes. Sequential incubation with oxidized LDL followed by acetylated LDL showed that the lipid-engorged lysosomes are long-lived structures, continuously accepting newly ingested lipoprotein. Comparison of acetylated and oxidized LDL in mouse peritoneal macrophages, a cell which does not accumulate substantial lysosomal lipid, also revealed no differences in uptake. This indicates that in THP-1 cells, the differences in metabolism of oxidized and acetylated LDL are due to cell-specific lysosomal or post-lysosomal events not present in B6C3F1 mouse macrophages. PMID- 12533239 TI - Unconventional specimen preparation techniques using high resolution low voltage field emission scanning electron microscopy to study cell motility, host cell invasion, and internal cell structures in Toxoplasma gondii. AB - Apicomplexan parasites employ complex and unconventional mechanisms for cell locomotion, host cell invasion, and cell division that are only poorly understood. While immunofluorescence and conventional transmission electron microscopy have been used to answer questions about the localization of some cytoskeletal proteins and cell organelles, many questions remain unanswered, partly because new methods are needed to study the complex interactions of cytoskeletal proteins and organelles that play a role in cell locomotion, host cell invasion, and cell division. The choice of fixation and preparation methods has proven critical for the analysis of cytoskeletal proteins because of the rapid turnover of actin filaments and the dense spatial organization of the cytoskeleton and its association with the complex membrane system. Here we introduce new methods to study structural aspects of cytoskeletal motility, host cell invasion, and cell division of Toxoplasma gondii, a most suitable laboratory model that is representative of apicomplexan parasites. The novel approach in our experiments is the use of high resolution low voltage field emission scanning electron microscopy (LVFESEM) combined with two new specimen preparation techniques. The first method uses LVFESEM after membrane extraction and stabilization of the cytoskeleton. This method allows viewing of actin filaments which had not been possible with any other method available so far. The second approach of imaging the parasite's ultrastructure and interactions with host cells uses semithick sections (200 nm) that are resin de-embedded (Ris and Malecki, 1993) and imaged with LVFESEM. This method allows analysis of structural detail in the parasite before and after host cell invasion and interactions with the membrane of the parasitophorous vacuole as well as parasite cell division. PMID- 12533240 TI - The canalicular structure of compact bone in the rat at different ages. AB - Osteocytes communicate through a canalicular system that maintains the vitality and mineral metabolism of bone. Casting the vascular canals and canaliculi of compact bone with methacrylate and viewing them with scanning electron microscopy shows their extent and relationships. Confocal laser scanning microscopy of the same specimen before corrosion establishes the degree of calcification of the different tissue components. These methods were used to compare basal with alveolar compact bone in the rat mandible at different ages. Sections of the mandibular molar region were placed in a methacrylate resin. After polymerization and study with confocal microscopy, the organic matrix was removed. Juvenile rats had large irregular central vascular canals and lacunae that were more concentric in the basal than the alveolar bone. Cast lacunae were round, and the canaliculi from these lacunae were short and thick in both bones. Adult rats had regular concentrically arranged lacunae in the basal bone. Cast lacunae were ellipsoid and flatter in the basal bone than in the alveolar bone. The intercommunicating canaliculi were increased and canaliculi had more branching than the juvenile rats. The aged rats had fewer vascular canals, lacunae, and canaliculi and had osteoporotic changes. The cast lacunae were slender and flat especially in the basal bone. The porosity of the mandible became more pronounced in the alveolar than in the basal bone with aging. The canaliculi of mandibular compact bone thinned and developed extensive branching with adulthood but decreased in size and number with advanced age. Lacunae proceed from the large circular structures of youth to the flat forms of the aged. These studies show that the internal structure of compact bone changes with age and mirrors its functional state. PMID- 12533241 TI - Structural aspects of the lymphocyte traffic in rat submandibular lymph node. AB - Modulation of lymphocyte flow in the lymphatic compartment of the lymph node may serve, in part, to promote lymphocyte sensitization during an antigenic challenge. This study was undertaken to show where this might occur by examining the structural relationships of the intranodal lymphatic pathways, blood vessels, and connective tissue support with respect to lymphocyte and lymph flow. Differently stained plastic resins were injected into the blood vessels and lymphatics of the submandibular lymph node and visualized with a confocal laser scanning microscope. The specimens were corroded to study the three-dimensional cast structures by scanning electron microscopy. Alkali digestion was also used to prepare the reticular fiber network in the lymph node for scanning electron microscopic examination. At the hilus of the node, two to three arteries gave off arterioles running in medullary cords towards the cortex. The medullary cords, the periphery of the deep cortex, and the perifollicular zones had dense capillary networks. In contrast, the center of the follicle and the center of the deep cortex were less highly vascularized. High-endothelial venules were restricted to the perifollicular zone and the periphery of the deep cortex. At the cortico-medullary boundary, they abruptly transformed into medullary venules with a normal endothelium. The marginal sinus of the lymph node was crossed by thick reticular fibers that arose from the inner sheets of the capsule. The lymph pathway went through the marginal sinus, into the trabecular sinus, to the cortical perifollicular sinus, the dense lymphatic sinus around the deep cortex, and finally into the medullary sinus. At present, the exact functional significance of the complex lymph node lymphatic architecture is not clear. However, the highly organized structural organization may play a significant role in regulating and directing lymphocyte flow to facilitate antigen presentation. PMID- 12533242 TI - New applications of electron diffraction in the pharmaceutical industry: polymorph determination by using a combination of electron diffraction and synchrotron X-ray powder diffraction techniques. AB - Electron diffraction has been recently used in the pharmaceutical industry to study the polymorphism in crystalline drug substances. While conventional X-ray diffraction patterns could not be used to determine the cell parameters of two forms of the microcrystalline GP IIb/IIIa receptor antagonist roxifiban, a combination of electron single-crystal and synchrotron powder diffraction techniques were able to clearly distinguish the two polymorphs. The unit-cell parameters of the two polymorphs were ultimately determined using new software routines designed to take advantage of each technique's unique capabilities. The combined use of transmission electron microscopy (TEM) and synchrotron patterns appears to be a good general approach for characterizing complex (low-symmetry, large-unit-cell, micron-sized) polymorphic pharmaceutical compounds. PMID- 12533243 TI - Quantitative modal determination of geological samples based on X-ray multielemental map acquisition. AB - Multielemental X-ray maps collected by a remote scanning system of the electron beam are processed by a dedicated software program performing accurate modal determination of geological samples. The classification of different mineral phases is based on elemental concentrations. The software program Petromod loads the maps into a database and computes a matrix consisting of numerical values proportional to the elemental concentrations. After an initial calibration, the program can perform the chemical composition calculated on the basis of a fixed number of oxygens for a selected area. In this way, it is possible to identify all the mineral phases occurring in the sample. Up to three elements can be selected to calculate the modal percentage of the identified mineral. An automated routine scans the whole set of maps and assigns each pixel that satisfies the imposed requirements to the selected phase. Repeating this procedure for every mineral phase occurring in the mapped area, a modal distribution of the rock-forming minerals can be performed. The final output consists of a digitized image, which can be further analyzed by common image analysis software, and a table containing the calculated modal percentages. The method is here applied to a volcanic and a metamorphic rock sample. PMID- 12533244 TI - Inhomogeneous vasodilatory responses of rat tail arteries to heat stress: evaluation by synchrotron radiation microangiography. AB - Tail blood flow is crucial for dissipating body heat in rats. Angiographies are convenient tools to evaluate tail circulation. However, conventional angiographies do not have sufficient sensitivity or spatial resolution for small vessels. Recently, we developed a novel microangiographic system using monochromatic synchrotron radiation and a high-definition video camera system. Here, we report an evaluation of rat tail circulation under heat stress using the synchrotron radiation microangiographic system. We performed an experiment using the microangiography of the caudal artery before and after heating up WKAH/HkmSlc rats to rectal temperature of 39 degrees C. The images were digitized and temporal subtraction was performed, and the diameters of caudal arteries were evaluated. After heating, the medial caudal artery was markedly dilated (320 +/- 53 to 853 +/- 243 micro m in diameter, p<0.001), while no significant change was observed in the lateral caudal arteries (139 +/- 42 to 167 +/- 73 micro m) and segmental anastomosing vessels. The heat stress allowed for visualization of the superficial caudal arteries with a diameter of approximately 60 micro m, not visible prior to heating. Thus, synchrotron radiation microangiography demonstrated that the rat tail possessed dual sets of arteries; one set was highly sensitive to heat-induced vasodilation (medial caudal artery and superficial caudal arteries) and the other set was less sensitive (lateral caudal arteries and segmental anastomosing vessels). PMID- 12533245 TI - Microvascular remodelling after endurance training with Co2+ treatment in the rat diaphragm and hind-leg muscles. AB - This study was designed to examine the changes in capillary geometry, especially the distribution of arteriolar and venular capillaries, in the skeletal muscles of female Wistar rats after endurance training with and without chronic CoCl(2) administration. Four groups of rats were used: non-treated sedentary, non-treated training, Co(2+)-treated sedentary, and Co(2+)-treated training. Exercise training by running lasted for 5 weeks at 25 m/min on a 20% gradient, 10-60 min/d, 5 d/week. The Co(2+)-treated rats drank water containing 0.01% CoCl(2) for 5 weeks. Morphological findings were obtained from the soleus (SOL), deep (PLd) and superficial (PLs) portions of plantaris, and diaphragm (DIA) muscles. Co(2+) administration significantly increased the blood hemoglobin concentration by approximately 25% with and without training. Only in DIA, the Co(2+) treatment alone significantly increased total capillary density and the capillary-to-fiber ratio (C : F) (p<0.05). Both training groups with and without Co(2+) administration showed a significant increase in the C : F in SOL and PLd (p<0.05). In PLd, the increase was significantly greater in the Co(2+)-treated training group than in the non-Co(2+)-treated training group (p<0.05). Training significantly increased the proportion of arteriolar capillaries while it decreased that of venular capillaries in both SOL and PLd (p<0.05). These changes were also observed in PLd after training with Co(2+). The densities of VEGF positive and TGF-beta1-positive capillaries remained unchanged in all muscle portions examined after either Co(2+) administration or exercise training. These results suggest that chronic Co(2+) administration causes adaptive changes in the oxygen transport system in respiratory muscle and facilitates exercise-induced angiogenesis in hind-leg muscles. PMID- 12533246 TI - Changes in blood pressure and muscle sympathetic nerve activity during water drinking in humans. AB - To investigate the possible involvement of the sympathetic nervous system in pressor response during water drinking, muscle sympathetic nerve activity (MSNA), blood pressure (BP), and heart rate (HR) were continuously measured in healthy young volunteers throughout the experiments of a 5-min control, 2 min of drinking 500 ml water, and a 28-min recovery. To avoid the effects of water passing through the oropharyngeal and esophageal regions and/or effects of swallowing, an equal amount of water was directly infused to the stomach through a stomach tube for 2 min. Water drinking caused a transient increase in mean arterial pressure (MAP) and HR immediately after drinking (DeltaMAP, 12.6 +/- 2.1 mmHg; DeltaHR, +19.9 +/- 1.7 beats/min at the peak). An abrupt decrease of MSNA was observed directly during water drinking (Deltaburst rate, -6.9 +/- 1.3 bursts/min; Deltatotal activity, -2,606 +/- 491 U/min), and it increased to the baseline level thereafter. Gastric infusion had little or no effect on MAP, HR, and MSNA. The present study demonstrated that a pressor response during water drinking was associated with the attenuation of MSNA and not generated by gastric infusion of water at the same rate as in this drinking manner. In conclusion, the rapid rise in BP might be caused through stimulations from the oropharyngeal region, swallowing-induced factors, and/or a feedforward mechanism by a central descending signal from the higher brain centers. PMID- 12533247 TI - Simultaneous optical and electrical recording of a single ion-channel. AB - In recent years, the single-molecule imaging technique has proven to be a valuable tool in solving many basic problems in biophysics. The technique used to measure single-molecule functions was initially developed to study electrophysiological properties of channel proteins. However, the technology to visualize single channels at work has not received as much attention. In this study, we have for the first time, simultaneously measured the optical and electrical properties of single-channel proteins. The large conductance calcium activated potassium channel (BK-channel) labeled with fluorescent dye molecules was incorporated into a planar bilayer membrane and the fluorescent image captured with a total internal reflection fluorescence microscope simultaneously with single-channel current recording. This innovative technology will greatly advance the study of channel proteins as well as signal transduction processes that involve ion permeation processes. PMID- 12533248 TI - The effects of losartan and enalapril therapies on the levels of nitric oxide, malondialdehyde, and glutathione in patients with essential hypertension. AB - Several recent studies have shown that essential hypertension is associated with increased oxidative stress, which may cause hypertension via enhanced oxidation and inactivation of nitric oxide. In this study, we investigated the malondialdehyde, nitric oxide, and glutathione levels in newly diagnosed essential hypertensive patients and whether or not there was any effect of antihypertensive treatment with angiotensin II type 1 receptor antagonist, losartan or angiotensin converting enzyme inhibitor, enalapril on plasma malondialdehyde, nitric oxide, and glutathione values. We selected 17 patients (F/M: 10/7, mean age: 46.12 +/- 9.2 years) for enalapril therapy (10-20 mg/d) and 14 patients (F/M: 8/6, mean age: 47.7 +/- 7.5 years) for losartan therapy (50-100 mg/d), and compared them with 12 normotensive controls. At the beginning of the study, both treated groups showed significantly higher plasma malondialdehyde and lower glutathione and nitric oxide in exhaled air compared to the control group. After 9 weeks of enalapril and losartan treatment, both systolic and diastolic pressure were significantly reduced. Both enalapril and losartan produced a significant decrease in plasma malondialdehyde and a significant increase in plasma glutathione levels and nitric oxide in exhaled air after 9 weeks. Initial values of plasma nitrate levels in patient groups were similar to the control group and increased significantly after the treatment period. In conclusion, both losartan and enalapril may be regulators between oxidant stress and the antioxidant system. PMID- 12533249 TI - Plasma-mediated potentiation in prostanoid-induced contractions in isolated canine external jugular veins. AB - We examined the effects of plasma on contractile responses of isolated dog external jugular veins to a thromboxane A(2) analog, U46619, and noradrenaline. Pretreatment with 1.0% plasma in Krebs-bicarbonate solution, but not 0.1%, caused a significant left and upward shift in the concentration-contractile response curve for U46619. The plasma-mediated potentiation of the response to U46619 was found in the venous segments without intact endothelium. The administration of 2x10(-5) M lysophosphatidylcholine in Krebs-bicarbonate solution with no plasma also produced a significant left and upward shift of the concentration contractile response curve for U46619, the shift being quite similar to that obtained with 1.0% plasma. In contrast, pretreatment with 1.0% plasma or 2x10(-5) M lysophosphatidylcholine produced no significant effect on the noradrenaline mediated contractions in the venous segments. Pretreatment with 10(-4) M L ascorbate or 0.1 mg/ml alpha-tocopherol in the presence of 1.0% plasma caused a significant reduction in the plasma-mediated potentiation of the contractile responses to U46619. These findings suggest that lysophosphatidylcholine, a major phospholipid component of oxidized low-density lipoproteins, may contribute, in part, to the plasma-mediated potentiation of contractile responses of the isolated veins to U46619, and that the antioxidant vitamin, L-ascorbate, or alpha tocopherol significantly reduces the plasma-mediated potentiation of the contractile responses to U46619, which may be related to inhibiting the production of lysophosphatidylcholine in plasma. PMID- 12533250 TI - Effect of zinc-carnosine complex on muscular function in frail distrophin deficient (mdx) mice. AB - The effect of zinc-carnosine complex (Z-103) on muscle function in dystrophin deficient (mdx) mice was examined using several different courses of repetitive administration. Z-103 at a dose of 100 mg/kg increased the load resistant time (LRT), during which an animal bearing a load holds himself upright on a wire net, in mdx mice when administered at an age of less than about 4 months. The effect of Z-103 on LRT was independent of sex when given by intraperitoneal (I.P.) administration between 4 and 8 weeks of age. Administration of Z-103 from the age of 4 to 9 weeks had no significant effect on wet weight, magnitude or rate of rise of twitch force, or rate of decay of twitch force over time with twitch elicited by 0.5 Hz of electricity in the extensor digitorum longus muscle or calcium content in the gastrocnemius muscle, while it increased the magnitude of twitch force in the soleus muscle. These results suggest that Z-103 reduces fatigability of the whole body in mdx mice, possibly by increasing the contractility of slow fibers. PMID- 12533251 TI - Inactivation of cardiac Na+ channel simply through open states as revealed by single-channel analysis in guinea pig ventricular myocytes. AB - Inactivation of the cardiac Na(+) channel was analyzed by recording channel currents from a cell-attached patch containing only one functional Na(+) channel in guinea-pig ventricular myocytes. A two-step test pulse, first to variable levels (Pulse 1) and then to -30 mV (Pulse 2) was applied from a holding potential of -140 mV. When a cumulative histogram was determined for the latency of first opening, the histogram was well fitted with a single exponential function at -70 to -30 mV of Pulse 1. The activation time course of ensemble average was virtually single exponential. Although the ensemble average of 500 sweeps showed various extents of inactivation during Pulse 1, the saturation level of the cumulative first-latency histogram at the end of the two-step pulse was almost constant (0.7-0.8), irrespective of Pulse 1. Even when the interval between successive test pulses was prolonged from 70 to 970 ms, the saturation level of the histogram was not modified. These findings are consistent with inactivation only through the open state. Thus, the apparent "blank sweep inactivation" does not necessarily indicate direct inactivation from closed states. These findings support the hypothesis that the inactivation of cardiac Na(+) channel occurs exclusively through the open state. PMID- 12533252 TI - Effects of cilostazol, a selective cyclic AMP phosphodiesterase inhibitor on isolated rabbit spinal arterioles. AB - Cilostazol, a potent inhibitor of guanosine 3':5'-cyclic monophosphate (cGMP) inhibited adenosine 3':5'-cyclic monophosphate (cAMP) phosphodiesterase (PDE3), has been used clinically for the treatment of chronic peripheral arterial occlusive disease. The beneficial effect of cilostazol is attributed to both anti platelet aggregating activity and vasodilation. However, the effect of cilostazol on resistance-sized vasculature is not well documented. Furthermore, mechanisms of vasodilation and influence on endothelium function are not fully understood. Thus, we investigated the vasodilator action of cilostazol using isolated, pressurized rabbit spinal arterioles with special reference to the functional endothelium. Cilostazol, acetylcholine (ACh), isocarbacyclin (prostacyclin analogue), and sodium nitroprusside (SNP) all produced concentration-dependent vasodilations of isolated spinal arterioles with endogenous myogenic tone. The order of potency of these agonists was isocarbacyclin>ACh>SNP>cilostazol. Indomethacin (10 micro M, a cyclo-oxygenase inhibitor), N(omega)-nitro-L-arginine methyl ester (L-NAME, a nitric oxide synthase inhibitor, 30 micro M), or chemical denudation of the endothelial cells did not significantly alter the cilostazol induced arteriolar dilation. Furthermore, stimulating the release of endothelium derived relaxing factors by administering ACh (100 nM), or treating with isocarbacyclin (1 nM) or SNP (3 nM) did not significantly modify the cilostazol induced vasodilation. These results suggest that cilostazol produces the vasodilation of isolated, pressurized rabbit spinal arterioles independent of the functional endothelium. We infer that the vasodilator action of cilostazol in the spinal arterioles may be attributed to a yet unknown mechanism that is independent of the PDE3 inhibition. PMID- 12533253 TI - Post-effects of nandrolone decanoate treatment on contractile responses of rat skeletal muscles. AB - The post-effects of nandrolone decanoate treatment (15 mg.kg(-1).week(-1)) were studied on contractile responses of isolated small bundles of intact slow (soleus) and fast-(extensor digitorum longus, edl) twitch fibers in rat. Five weeks of treatment induced, in edl, an increase in the amplitude of twitch (55%) and K(+) contracture (32%) without significant change in the time constant of relaxation and caffeine contracture. In soleus, an increase in the amplitude of twitch (35%) and caffeine contracture (0.2 mM: 218%, 0.5 mM: 88%, 5 mM: 28%, and 10 mM: 25%) was observed without change in K(+) contracture characteristics. In addition to these effects, 10 weeks of drug treatment increased the amplitude of soleus K(+) contracture (29%) and edl caffeine contracture (0.2 mM: 247%, 0.5 mM: 170%, 5 mM: 29%, and 10 mM: 45%), and reduced the 50% recovery time for K(+) contracture (EC(50)) (soleus: 37%, edl: 12%). After 5-week treatment followed by 5-week arrest, as compared to the control group, no change in the amplitude of twitch, K(+) or caffeine contractures was found in edl. In soleus, although no significant difference was observed in the amplitude of twitch, the amplitude of K(+) (35%) and caffeine contractures (0.2 mM: 227%, 0.5 mM: 128%) remained greater than the control group. Moreover, the EC(50) values were prolonged (46%) in soleus, whereas no significant difference was observed in edl. The present work suggests that nandrolone decanoate treatment induced differential post effects on contractile responses developed in slow- and fast-twitch skeletal muscles by acting differently on the different steps of the excitation contraction coupling mechanism. PMID- 12533254 TI - VE response to VCO2 during exercise is unaffected by exercise training and different exercise limbs. AB - We designed two experiments to investigate the relationship between ventilation (VE) and CO2 output (VCO2) during exercise under the conditions of exercising different limbs, the arms as opposed to the legs (experiment 1), and of different physical training states after undergoing standard exercise training for 90 d (experiment 2). Six healthy young subjects underwent submaximal ramp exercise at an incremental work rate of 15 W/min for the arm and leg, and 11 healthy middle aged subjects underwent an incremental exercise test at the rate of 30 W/3 min before and after exercise training. We measured pulmonary breath-by-breath VE, VCO2, oxygen uptake (VO2), tidal volume (VT), breathing frequency (bf), and end tidal O2 and CO2 pressures (PETO2, PETCO2) via a computerized metabolic cart. In experiment 1, arm exercise produced significantly greater VE than did leg exercise at the same work rates, as well as significantly higher VO2, VCO2, and bf. The slopes of the regression lines in the VE-VCO2 relationship were not significantly different: the values were 27.8 +/- 2.1 (SD) during the arm exercise, and 25.3 +/- 3.9 during the leg exercise, with no differences in their intercepts. In experiment 2, the VO2, VCO2, and VE responses at the same work rates were similar in both before and after the 90-d exercise training, whereas the heart rate (HR) and mean blood pressure (MBP) were significantly reduced after training. Exercise training did not alter the VE-VCO2 relationship, the slope of which was 31.9 +/- 4.9 before exercise training and 34.2 +/- 4.4 after exercise training. We concluded that the VE-VCO2 relationship during exercise is unaltered, independent of not only working muscle regions but also exercise training states. PMID- 12533255 TI - Convenient automated conductance volumetric system. AB - Conventional conductance volumetric systems require ex-vivo calibrations for blood conductivity and parallel conductance. It is often impractical to repeat blood sampling and hypertonic saline infusion for these calibrations. To overcome these limitations, we developed a useful, self-calibrating conductance volumetric system that does not require ex-vivo calibrations. On a conventional 6-electrode catheter, we added an extra electrode close to one of the recording electrodes to estimate blood conductivity. These two electrodes were placed close (0.5 mm) enough so that conductance between them reflected only blood conductivity regardless of cardiac volume. We estimated parallel conductance by the dual frequency excitation (2 and 20 kHz) method. In 18 anesthetized rabbits, blood conductivity (sigma(est)) thus estimated agreed well with that (sigma(conv)) measured by the conventional ex-vivo blood sampling method (sigma(est) = 1.04sigma(conv)-0.25, R(2) = 0.98, SEE = 0.01 mS/cm, 1.2% error). Parallel conductance (G(p est)) estimated by dual-frequency excitation also agreed well with that (G(p conv)) estimated by the saline injection method (G(p est) = 0.95G(p conv)+4.25, R(2) = 0.87, SEE = 4.0 mS, 6.0% error). Estimated ventricular volume (V(est)) by our system agreed reasonably well with that (V(conv)) by the conventional method (V(est) = 0.93V(conv)+0.01, R(2) = 0.86, SEE = 0.22 ml, 14.7% error). The fact that this self-calibrating conductance volumetric system drastically simplifies volume measurement makes it an attractive tool for the assessment of cardiac function where significant changes in blood conductivity and parallel conductance are inevitable, such as in cardiac surgery. PMID- 12533256 TI - Local control of breast cancer: issues and answers. PMID- 12533260 TI - Will we need lymph node dissection at all in the future? AB - Traditionally in the treatment of primary breast cancer, axillary lymph node dissection (ALND) plays an important role. However, a substantial and increasing percentage of patients appear to have no nodal involvement and have been subjected to ALND unnecessarily. The first reason to perform an ALND is axillary nodal staging. After reviewing the literature, it can be concluded that in clinically node-negative patients an adequately conducted lymphatic mapping by sentinel node procedure is equal to ALND for this purpose. The second reason to perform an ALND is to establish the extent of nodal involvement, which might have an impact on adjuvant treatment recommendations. However, there is no evidence available that patients with extensive nodal involvement (= 4 positive nodes) benefit more from adjuvant systemic treatment (either standard or high dose) in terms of reduction of odds of recurrence and mortality compared to patients with limited nodal involvement and optimally administered so-called standard adjuvant treatment. The third reason to perform an ALND is to ensure axillary tumor control. Reviewing the different treatment options, it can be concluded that in clinically node-negative patients axillary control after axillary radiotherapy appears to be similar to axillary control after ALND. In clinically overt axillary involvement, ALND (with or without adjuvant radiotherapy) may result in an improved regional control. In the near future, ALND will not be the standard of care but will be reserved for those patients with proven axillary lymph node involvement. In microscopic disease, radiotherapy may be an alternative with equal control and less morbidity. PMID- 12533262 TI - Adjuvant use of taxanes for patients with breast cancer: we see the tip of the iceberg. AB - Advances in screening techniques for breast cancer have led to the diagnosis of more patients at earlier disease stages at which time the possibility of a cure is more likely. Adjuvant chemotherapy with anthracycline-based regimens has proven to reduce the risk of relapse and cancer-related death in women with early stage breast cancer. Recent studies have aimed at integrating the taxanes, paclitaxel and docetaxel, into the adjuvant setting, but to date, we are still in the earliest stages of the study of patients with operable breast cancer. Adjuvant trials now require thousands of patients and many years to reach maturity. Many of the trials began in the late 1990s and are not yet mature. For node-positive patients, the available evidence supports the use of taxanes as adjuvant treatment since they are safe and appear to provide benefit. Going forward, docetaxel holds significant promise in the adjuvant setting, and further trials as well as further follow-up of existing trials are eagerly awaited to help us determine whether docetaxel is best given sequentially to, or concurrently with, doxorubicin or epirubicin. PMID- 12533263 TI - Phase II trial of doxorubicin/docetaxel/cyclophosphamide for locally advanced and metastatic breast cancer: results from NSABP trial BP-58. AB - Based on the recommended phase II doses for doxorubicin (60 mg/m2) and docetaxel (60 mg/m2) and the National Surgical Adjuvant Breast and Bowel Project's (NSABP) experience with doxorubicin and cyclophosphamide (cyclophosphamide 600 mg/m2), we conducted a phase II trial at 18 institutions using doxorubicin/docetaxel/cyclophosphamide (ATC) given every 21 days, in preparation for a major adjuvant breast cancer study (NSABP B-30), in which ATC would be used. Eligibility requirements included measurable stage IIIB/IV breast cancer, performance status 0-2, normal left ventricular ejection fraction, and no prior chemotherapy for metastatic disease (nontaxane adjuvant chemotherapy was allowed if completed > 12 months before entry and if the cumulative dose of doxorubicin was =240 mg/m2). Eighty-nine patients were entered who ranged in age from 30-78 years (38.2% < 50 years; 61.8% =50 years). A total of 33.7% of patients had stage IIIB disease, and 66.3% had stage IV disease. Among the stage IV patients, 20.3% had received prior adjuvant chemotherapy. Dexamethasone premedication (8 mg p.o. b.i.d. for 3 days) and prophylactic ciprofloxacin (500 mg p.o. b.i.d. days 5-15) were used. Colony-stimulating growth factors were reserved for secondary prophylaxis after prolonged or febrile neutropenia (FN) or documented severe infection in a prior cycle. After a cumulative dose of doxorubicin 480 mg/m2, patients could continue with docetaxel/cyclophosphamide alone. Eighty-nine patients and 577 courses were evaluable for toxicity. Median time on study as of May 2002 was 36.5 months (range, 28-47 months). Febrile neutropenia occurred in 34 patients (38%); 8 developed FN in the absence of prior prophylactic growth factor support; 26 developed FN despite prior growth factor support (for one patient this information was unavailable). There were no septic deaths. One patient died from pulmonary embolism. Other grade 3/4 adverse events included: nausea (9%), vomiting (7%), stomatitis (6%), diarrhea (4%), arthralgia/myalgia (3%), and neurotoxicity (1%). Clinical congestive heart failure was seen in 3 patients (3.4%). Seventy-seven patients were evaluable for best response within 6 cycles of therapy. Thirteen patients (16.9%) had a complete response, 43 (55.8%) had a partial response, for an overall response rate of 72.7%. The median response duration was 23.8 months (95% CI, 16.2-37.8 months), and the median time to progression or death was 23.5 months (95% CI, 16.3-38.7 months). The median survival time was 35.6 months (95% CI, 26.6-39.4 months). The administration of ATC with primary ciprofloxacin and secondary colony-stimulating factor prophylaxis is feasible and active. Its value in the adjuvant setting is currently under investigation. PMID- 12533264 TI - Microtubule-associated parameters as predictive markers of docetaxel activity in advanced breast cancer patients: results of a pilot study. AB - Docetaxel is currently one of the most active agents for breast cancer. Predictive markers of docetaxel efficacy are clearly needed in order to avoid unnecessary toxicity in nonresponding or resistant patients and to improve the cost-effectiveness ratio of docetaxel. This pilot study correlates the clinical efficacy of docetaxel in 54 metastatic or locally advanced breast cancer patients with the expression of microtubule-associated parameters evaluated by immunohistochemistry in archival tumor samples. Among the 41 eligible patients (evaluable response to docetaxel and available predocetaxel treatment paraffin embedded tumor tissue), response to docetaxel was: partial response 54%, stable disease 29%, and progressive disease 17%. Alfa- and b-tubulin and Tau protein were expressed in the majority of tumor samples. Class II, III, and IV b-tubulin isotypes were expressed in 56%, 65%, and 82% of samples, respectively. No clear association was found between response to docetaxel and the level of expression of Tau protein, a- and b-tubulin, and class III and IV b-tubulin isotypes. In patients with class II b-tubulin-positive tumors, the response rate was 39%, while in class II b-tubulin-negative tumors the response rate was 79% (P = 0.04). Therefore, we conclude that the class II b-tubulin isotype seems to be a promising predictive marker of docetaxel activity. Nevertheless, further investigations are needed due to the limited number of patients evaluated in this pilot study. PMID- 12533265 TI - Weekly docetaxel/paclitaxel in pretreated metastatic breast cancer. AB - The purpose of our study was to evaluate the feasibility and efficacy of weekly docetaxel/paclitaxel in pretreated advanced breast cancer patients. Twenty-six patients with metastatic breast cancer were included in this study. Three different schedules of treatment were administered. The starting schedule, A1, consisted of docetaxel 60 mg/m2 on day 1 plus paclitaxel 60 mg/m2 over 1 hour, weekly for 18 weeks; this schedule was considered feasible if at least 70% of the planned doses were given on time and without reduction. Schedule A2 consisted of the same doses administered on days 1 and 8 every 3 weeks, and schedule B consisted of docetaxel 25 mg/m2 followed by paclitaxel 40 mg/m2 for 1 hour on days 1 and 8 every 3 weeks for a total of 6 cycles. All patients had received prior anthracyclines, and 19 patients were pretreated with taxanes. Seventy-seven percent of patients had received at least 2 prior lines of chemotherapy. Twenty five patients are assessable for toxicity and efficacy. A total of 109 cycles of chemotherapy have been administered, with a median of 4 cycles per patient (range, 1-8 cycles). The median delivered dose intensity was 27 mg/m2/week for paclitaxel (range, 18-50 mg/m2/week) and 17 mg/m2/week (range, 12-39 mg/m2/week) for docetaxel. Six patients received schedule A1. This schedule was considered not feasible due to neutropenia grade > 2, mucositis, and diarrhea grade 2, which required dose reduction/omission in 33% of administrations. For this reason, treatment in the following 5 patients was omitted on day 15 (schedule A2). Schedule B was found to be more feasible with 16% of dose reductions/omissions. The overall response rate was 68% (95% CI, 50%-86%) with a median duration of response of 10 months (range, 2-18+ months). Treatment was well tolerated; myelosuppression was rare and grade 3 cutaneous toxicity was observed in only 2 patients. In conclusion, weekly docetaxel/paclitaxel is active at low dosages and was well tolerated as salvage chemotherapy in metastatic breast cancer. This regimen represents a valid option as a salvage treatment in taxane- and anthracycline-pretreated patients. PMID- 12533266 TI - Toward re-expressing tumor suppresor gene maspin in breast cancer. PMID- 12533268 TI - Breast cancer chemotherapy-related cognitive dysfunction. AB - Cognitive side effects of systemic chemotherapy have become an increasing concern among breast cancer survivors, their families, and health care professionals. A growing body of research supports the hypothesis that chemotherapy can produce long-term cognitive changes in at least a subgroup of cancer survivors. We review evidence implicating systemic chemotherapy as the cause of cognitive changes; describe the limitations due to lack of longitudinal studies and gaps in knowledge (ie, no clear mechanism by which chemotherapy can produce cognitive changes has been proposed); discuss possible factors like age, intelligence quotient/education, and psychological, genetic, and hormonal factors that might increase risk for chemotherapy-induced cognitive changes; and outline future directions for research. Such future research includes large-scale, longitudinal studies of pretreatment neuropsychological assessments, use of imaging techniques and the development of animal models to study the mechanisms of chemotherapy induced changes in cognitive functioning, and the development of interventions to prevent or reduce the negative cognitive effects of chemotherapy PMID- 12533269 TI - Assessing cognitive dysfunction in breast cancer: what are the tools? AB - The goal of adjuvant chemotherapy in the treatment of breast cancer is to reduce recurrence and mortality. With respect to quality of life and morbidity, however, such treatments come at a cost. Decreased cognitive functioning, development of fatigue, and mood alterations are common during chemotherapy and persist after its conclusion as evidenced by subjective self-reports and objective neurocognitive performance records. Few efforts, however, have used standardized neuropsychological measures, and no study has empirically selected those measures that best distinguish women in active chemotherapy from those who have previously completed it. Perhaps the most glaring deficit in the literature is that no study has used baseline data to track individual neurocognitive changes across treatment phases and after completion. This article provides an overview of the field of neuropsychology and the cognitive domains theorized to be affected by chemotherapy and the measures typically used, including validated computerized tests, which are tools for future studies; briefly summarizes existing research on the cognitive effects that chemotherapy has on breast cancer patients; compares data resulting from an ongoing pilot study of the cognitive performance of women actively undergoing anthracycline-containing chemotherapy with that of women 6-12 months post chemotherapy completion; and provides a preliminary analysis of the relationship between cognitive and emotional functioning. Future uses of these data to refine the ideal tools that efficiently, accurately, and validly detect short-term and persistent chemotherapy effects are proposed. PMID- 12533270 TI - Cognitive dysfunction and chemotherapy: neuropsychological findings in perspective. AB - Currently, the interest in cognitive functioning following chemotherapy is rapidly expanding as is reflected in a growing number of published studies on this topic. Although most studies are indicative of cognitive deficits after chemotherapy, definite conclusions on the role of chemotherapy on cognitive function can often not be drawn due to methodological problems. On the basis of the studies on cognitive functioning after chemotherapy that are conducted in The Netherlands Cancer Institute, the current article describes a number of such methodological topics that obscure straightforward interpretation of neuropsychological findings in toxicity research. Measurement issues that diverge from usual assessment issues encountered in psychosocial oncology will be described, and factors that might play a role in the cause of cognitive impairment will be evaluated. Also, future developments necessary to gain more insight into the prevalence, the pattern, and the impact of cognitive problems following chemotherapy are discussed PMID- 12533271 TI - Recombinant human erythropoietin for neuroprotection: what is the evidence? AB - Erythropoietin (EPO) is an endogenous cytokine with antiapoptotic, antiinflammatory, and neurotrophic properties. Apart from being produced by the kidney, liver, and spleen in response to hypoxia, EPO is highly expressed in the brain during development and after neuropathological insults. The observation that receptors for EPO are present on brain capillaries and glial capillary end feet has suggested that circulating (plasma) EPO may be transferred into the brain. This review summarizes the increasing number of studies indicating that peripherally administered recombinant human (rHu) EPO crosses the blood-brain barrier. Moreover, several of these studies have shown that peripherally administered rHuEPO can protect against the damage caused by a diversity of neuropathological conditions such as (a) stroke, (b) head and spinal cord trauma, (c) inflammatory and demyelinating conditions, (d) toxin-induced epileptic seizures, and (e) retinal ischemia. While all these studies are based on experiments in animal models, the effectiveness of rHuEPO in ischemic stroke in human patients has recently been suggested in a proof-of-concept trial, which is also discussed. PMID- 12533272 TI - Effects of epoetin alfa on cognitive function, mood, asthenia, and quality of life in women with breast cancer undergoing adjuvant chemotherapy. AB - Several recently published studies describe moderate to severe cognitive dysfunction in breast cancer survivors who were treated with adjuvant chemotherapy 1-5 years before undergoing extensive neuropsychological testing. While these studies are hypothesis-generating and preliminary given their small size and retrospective nature, they consistently suggest that between approximately 15% and 25% of chemotherapy-treated breast cancer patients will have evidence of cognitive dysfunction some years after chemotherapy, compared to about 10% of breast cancer survivors who did not receive chemotherapy. Recent preclinical data strongly suggest that erythropoetin is a potent, endogenous neuroprotective agent that prevents neuronal apoptosis from a variety of insults including hypoxia, trauma, subarachnoidal hemorrhage, and encephalitis. Erythropoietin also appears to enhance learning in a mouse spatial learning maze model. We have conducted a pilot study of epoetin alfa versus placebo in early stage breast cancer patients who received standard adjuvant anthracycline-based chemotherapy to determine the feasibility of administering standardized neurocognitive assessment tests in the oncology practice setting in order to understand whether the Executive Interview 25 test can detect the subtle cognitive impairment in verbal fluency, attention, and short-term memory observed with chemotherapy, and to assess whether epoetin alfa-treated patients have less evidence of cognitive dysfunction during and 6 months after chemotherapy compared with control-treated patients. We report here the preliminary results of this pilot clinical trial. PMID- 12533273 TI - Novel approaches to preventing chemotherapy-induced cognitive dysfunction in breast cancer: the art of the possible. AB - Chemotherapy-induced cognitive dysfunction in women with breast cancer has become an increasingly important clinical issue. To date, little is known about its incidence, exact characteristics, and exact pathophysiology. Likewise, no treatments have been shown to prevent or decrease cognitive changes thought to result from chemotherapy. However, ongoing scientific research might help us understand the mechanisms that will help patients maintain maximal cognitive function. Changes in cognition due to chemotherapy might result from indirect chemical toxicity and oxidative damage, direct injury to neurons, inflammation, or a type of autoimmune response. Based on these potential causes, and based on interventions that have been tested in dementia and Alzheimer's disease, there are a number of potential, novel interventions that could be tested in clinical trials as treatments for chemotherapy-induced cognitive dysfunction. Possible anecdotal strategies to consider include hormonal interventions, antioxidants, monoamine oxidase inhibitors, growth factors, dopamine agonists, cholinesterase inhibitors, antiinflammatory agents, and behavioral interventions PMID- 12533274 TI - Maintaining fluoroquinolone class efficacy: review of influencing factors. AB - Previous experience with antimicrobial resistance has emphasized the importance of appropriate stewardship of these pharmacotherapeutic agents. The introduction of fluoroquinolones provided potent new drugs directed primarily against gram negative pathogens, while the newer members of this class demonstrate more activity against gram-positive species, including Streptococcus pneumoniae. Although these agents are clinically effective against a broad range of infectious agents, emergence of resistance and associated clinical failures have prompted reexamination of their use. Appropriate use revolves around two key objectives: 1) only prescribing antimicrobial therapy when it is beneficial and 2) using the agents with optimal activity against the expected pathogens. Pharmacodynamic principles and properties can be applied to achieve the latter objective when prescribing agents belonging to the fluoroquinolone class. A focused approach emphasizing "correct-spectrum" coverage may reduce development of antimicrobial resistance and maintain class efficacy. PMID- 12533276 TI - Geographic translocation of bats: known and potential problems. AB - Natural, accidental, and intentional translocation of bats, both intra- and intercontinentally, has been documented. Some bats have been translocated while incubating infectious diseases, including rabies or related lyssavirus infections; others have escaped confinement en route to or at their destinations, while others have been released deliberately. Known events and potential consequences of bat translocation are reviewed, including a proposed solution to the attendant problems. PMID- 12533275 TI - Dead bugs don't mutate: susceptibility issues in the emergence of bacterial resistance. AB - The global emergence of antibacterial resistance among common and atypical respiratory pathogens in the last decade necessitates the strategic application of antibacterial agents. The use of bactericidal rather than bacteriostatic agents as first-line therapy is recommended because the eradication of microorganisms serves to curtail, although not avoid, the development of bacterial resistance. Bactericidal activity is achieved with specific classes of antimicrobial agents as well as by combination therapy. Newer classes of antibacterial agents, such as the fluoroquinolones and certain members of the macrolide/lincosamine/streptogramin class have increased bactericidal activity compared with traditional agents. More recently, the ketolides (novel, semisynthetic, erythromycin-A derivatives) have demonstrated potent bactericidal activity against key respiratory pathogens, including Streptococcus pneumoniae, Haemophilus influenzae, Chlamydia pneumoniae, and Moraxella catarrhalis. Moreover, the ketolides are associated with a low potential for inducing resistance, making them promising first-line agents for respiratory tract infections. PMID- 12533277 TI - Foot and mouth disease in livestock and reduced cryptosporidiosis in humans, England and Wales. AB - During the 2001 epidemic of foot and mouth disease (FMD) in livestock in England and Wales, we discovered a corresponding decrease in laboratory reports of cryptosporidiosis in humans. Using a regression model of laboratory reports of cryptosporidiosis, we found an estimated 35% (95% confidence interval [CI] 20% to 47%) reduction in reports during the weeks spanning the period from the first and last cases of FMD. The largest reduction occurred in northwest England, where the estimated decrease was 63% (95% CI 31% to 80%). Genotyping a subgroup of human isolates suggested that the proportion of Cryptosporidium genotype 2 strain (animal and human) was lower during the weeks of the FMD epidemic in 2001 compared with the same weeks in 2000. Our observations are consistent with livestock making a substantial contribution to Cryptosporidium infection in humans in England and Wales; our findings have implications for agriculture, visitors to rural areas, water companies, and regulators. PMID- 12533278 TI - Congenital transmission of Trypanosoma cruzi infection in Argentina. AB - Trypanosoma cruzi, the causative agent of Chagas disease, infects 10-18 million people and may be transmitted to the newborn. Using various data sources, we estimated that nearly 850 congenital cases occurred in Argentina in 1993, or 6.3 expected cases per each reported case in 1994 and in 1994-2001. The congenital transmission of T. cruzi constitutes a sizeable public health problem in the region. PMID- 12533279 TI - A molecular surveillance system for global patterns of drug resistance in imported malaria. AB - Analysis of imported malaria in travelers may represent a novel surveillance system for drug-resistant malaria. We analyzed consecutive falciparum malaria isolates from Canadian travelers from 1994 to 2000, for polymorphisms in pfcrt, dhfr, and dhps linked to chloroquine and pyrimethamine/sulfadoxine resistance. Forty percent of isolates possessed the K76 pfcrt allele, suggesting that many imported falciparum infections are still responsive to chloroquine. Travelers who had recently taken chloroquine had a significantly increased risk of harboring isolates with pfcrt resistance alleles (odds ratio = 4.47; p=0.03). The presence of two or more mutations in dhfr or dhps was found in 64.8% (95% confidence interval [CI] 54.6 to 73.9) and in 30.4% (95% CI 21.7 to 40.3) of isolates, respectively, and increased significantly over the course of the study. These molecular markers indicate that pyrimethamine/sulfadoxine resistance is increasing and is now too high to rely on this drug as a routine therapeutic agent to treat malaria in travelers. PMID- 12533280 TI - Cost-effectiveness of a potential vaccine for human papillomavirus. AB - Human papillomavirus (HPV) infection, usually a sexually transmitted disease, is a risk factor for cervical cancer. Given the substantial disease and death associated with HPV and cervical cancer, development of a prophylactic HPV vaccine is a public health priority. We evaluated the cost-effectiveness of vaccinating adolescent girls for high-risk HPV infections relative to current practice. A vaccine with a 75% probability of immunity against high-risk HPV infection resulted in a life-expectancy gain of 2.8 days or 4.0 quality-adjusted life days at a cost of $246 relative to current practice (incremental cost effectiveness of $22,755/quality-adjusted life year [QALY]). If all 12-year-old girls currently living in the United States were vaccinated, >1,300 deaths from cervical cancer would be averted during their lifetimes. Vaccination of girls against high-risk HPV is relatively cost effective even when vaccine efficacy is low. If the vaccine efficacy rate is 35%, the cost effectiveness increases to $52,398/QALY. Although gains in life expectancy may be modest at the individual level, population benefits are substantial. PMID- 12533281 TI - Natural enzootic vectors of Venezuelan equine encephalitis virus, Magdalena Valley, Colombia. AB - To characterize the transmission cycle of enzootic Venezuelan equine encephalitis virus (VEEV) strains believed to represent an epizootic progenitor, we identified natural vectors in a sylvatic focus in the middle Magdalena Valley of Colombia. Hamster-baited traps were placed into an active forest focus, and mosquitoes collected from each trap in which a hamster became infected were sorted by species and assayed for virus. In 18 cases, a single, initial, high-titered mosquito pool representing the vector species was identified. These vectors included Culex (Melanoconion) vomerifer (11 transmission events), Cx. (Mel.) pedroi (5 transmissions) and Cx. (Mel.) adamesi (2 transmissions). These results extend the number of proven enzootic VEEV vectors to 7, all of which are members of the Spissipes section of the subgenus Melanoconion. Our findings contrast with previous studies, which have indicated that a single species usually serves as the principal enzootic VEEV vector at a given location. PMID- 12533282 TI - Highly endemic, waterborne toxoplasmosis in north Rio de Janeiro state, Brazil. AB - In Campos dos Goytacazes, northern Rio de Janeiro state, Brazil, reports of uveitis consistent with toxoplasmosis led to a survey of the prevalence and risk factors for Toxoplasma gondii infection in 1997-1999. The survey population was selected randomly from schools, randomly chosen communities, and an army battalion. Serum samples from 1,436 persons were tested. With results adjusted for age, 84% of the population in the lower socioeconomic group was seropositive, compared with 62% and 23% of the middle and upper socioeconomic groups, respectively (p<0.001). When multivariate analysis was performed, drinking unfiltered water was found to increase the risk of seropositivity for the lower socioeconomic (odds ratio [OR]: 3.0, 95% confidence interval [CI] 1.3 to 6.9) and middle socioeconomic (OR: 1.7, 95% CI 1.2 to 2.3) populations. We also found a high T. gondii seroprevalence in this Brazilian community. Drinking unfiltered water increased the risk of T. gondii seropositivity, indicating the potential importance of oocyst transmission in water in this region. PMID- 12533283 TI - Seasonal dynamics of Anaplasma phagocytophila in a rodent-tick (Ixodes trianguliceps) system, United Kingdom. AB - We investigated the reservoir role of European wild rodents for Anaplasma phagocytophila using polymerase chain reaction (PCR) analysis of blood collected from individually tagged rodents captured monthly over 2 years. The only tick species observed in the woodland study site was Ixodes trianguliceps, and ruminant reservoir hosts were not known to occur. A. phagocytophila infections were detected in both bank voles and wood mice but were restricted to periods of peak nymphal and adult tick activity. Most PCR-positive rodents were positive only once, suggesting that rodent infections are generally short-lived and that ticks rather than rodents may maintain the infection over winter. Bank voles were more likely to be PCR positive than wood mice, possibly because detectable infections are longer lived in bank voles. This study confirms that woodland rodents can maintain A. phagocytophila in Great Britain in the absence of other reservoir hosts and suggests that I. trianguliceps is a competent vector. PMID- 12533284 TI - Two epidemiologic patterns of norovirus outbreaks: surveillance in England and wales, 1992-2000. AB - In the period 1992-2000, the Public Health Laboratory Service Communicable Disease Surveillance Centre collected standardized epidemiologic data on 1,877 general outbreaks of Norovirus (formerly "Norwalk-like virus") infection in England and Wales. Seventy-nine percent of general outbreaks occurred in health care institutions, i.e., hospitals (40%) and residential-care facilities (39%). When compared with outbreaks in other settings, those in health-care institutions were unique in exhibiting a winter peak (p<0.0001); these outbreaks were also associated with significantly higher death rates and prolonged duration but were smaller in size and less likely to be foodborne. These data suggest that Norovirus infection has considerable impact on the health service and the vulnerable populations residing in institutions such as hospitals and residential homes. A distinct outbreak pattern in health-care institutions suggests a combination of host, virologic, and environmental factors that mediate these divergent epidemiologic patterns. PMID- 12533285 TI - Epidemic hand, foot and mouth disease caused by human enterovirus 71, Singapore. AB - Singapore experienced a large epidemic of hand, foot and mouth disease (HFMD) in 2000. After reviewing HFMD notifications from doctors and child-care centers, we found that the incidence of HFMD rose in September and declined at the end of October. During this period, 3,790 cases were reported. We performed enteroviral cultures on 311 and 157 specimens from 175 HFMD patients and 107 non-HFMD patients, respectively; human enterovirus 71 (HEV71) was the most frequently isolated virus from both groups. Most of the HFMD patients were 85% (N =3,714) of all nonbacterial outbreaks of gastroenteritis reported from 1995 to 2000. However, the absolute number and population-based rates of viral gastroenteritis outbreaks differed markedly among European surveillance systems. A wide range of estimates of the importance of foodborne transmission were also found. We review these differences within the context of the sources of outbreak surveillance information, clinical definitions, and structures of the outbreak surveillance systems. PMID- 12533288 TI - The U.S.-Mexico Border Infectious Disease Surveillance project: establishing bi national border surveillance. AB - In 1997, the Centers for Disease Control and Prevention, the Mexican Secretariat of Health, and border health officials began the development of the Border Infectious Disease Surveillance (BIDS) project, a surveillance system for infectious diseases along the U.S.-Mexico border. During a 3-year period, a binational team implemented an active, sentinel surveillance system for hepatitis and febrile exanthems at 13 clinical sites. The network developed surveillance protocols, trained nine surveillance coordinators, established serologic testing at four Mexican border laboratories, and created agreements for data sharing and notification of selected diseases and outbreaks. BIDS facilitated investigations of dengue fever in Texas-Tamaulipas and measles in California-Baja California. BIDS demonstrates that a binational effort with local, state, and federal participation can create a regional surveillance system that crosses an international border. Reducing administrative, infrastructure, and political barriers to cross-border public health collaboration will enhance the effectiveness of disease prevention projects such as BIDS. PMID- 12533289 TI - Chagas disease in a domestic transmission cycle, southern Texas, USA. AB - After three dogs died from acute Chagas cardiomyopathy at one location, an investigation was conducted of the home, garage, and grounds of the owner. A serologic study was conducted on stray dogs, and an ecologic niche model was developed to predict areas where the vector Triatoma gerstaeckeri might be expected. PMID- 12533290 TI - Shiga toxin-producing Escherichia coli infections associated with hemolytic uremic syndrome, Italy, 1988-2000. AB - The mean annual incidence of hemolytic uremic syndrome in persons 6 million animals were slaughtered, and strict restrictions on access to the countryside were imposed. We report a dramatic decline in the reported incidence of human cryptosporidiosis in northwest England during weeks 13-38 in 2001, compared with the previous 11 years. This decline coincided with the period of foot and mouth restrictions. No similar reduction occurred in the other 26 weeks of the year. We also noted a substantial decline in the proportion of human infections caused by the bovine strain (genotype 2) of Cryptosporidium parvum during weeks 13-38 in that year but not during the other weeks. PMID- 12533292 TI - Enteropathogenic Escherichia coli O157 strains from Brazil. AB - We describe two serogroup O157 Escherichia coli strains from Brazilian infants with diarrhea. A variety of assays indicate that these strains belong to the enteropathogenic, not the enterohemorrhagic, pathotype. These strains possess a novel bfpA allele encoding the type IV pilin characteristic of typical enteropathogenic E. coli strains. Our results emphasize the pitfalls of classifying pathogenic E. coli by serogroup. PMID- 12533293 TI - Transfusion-associated babesiosis after heart transplant. AB - We describe a 54-year-old spleen-intact man with transfusion-associated Babesia microti infection after a heart transplant. Adult respiratory distress syndrome developed in the patient, and he required mechanical ventilation. Our experiences with this patient suggest that babesiosis should be considered in the differential diagnosis of transplant patients who have fever and hemolytic anemia. PMID- 12533294 TI - Household contamination with Salmonella enterica. AB - Household contamination with Salmonella enterica increases when occupational exposure exists (cattle farms with known salmonellosis in cattle, a salmonella research laboratory, or a veterinary clinic experiencing an outbreak of salmonellosis). Fifteen of 55 (27.2%) vacuum cleaner bags from households with occupational exposure to S. enterica were positive versus 1 of 24 (4.2%) without known exposure. Use of a carpet cleaner and several cleaners/disinfectants reduced, but failed to eliminate, S. enterica from artificially contaminated carpet. PMID- 12533295 TI - Persistence of W135 Neisseria meningitidis carriage in returning Hajj pilgrims: risk for early and late transmission to household contacts. AB - After an outbreak of meningococcal disease caused by Neisseria meningitidis W135, associated with the Hajj pilgrimage in 2001, 15% of returning vaccinated pilgrims carried a single W135 clone, and 55% were still carriers 6 months later. Transmission to 8% of their unvaccinated household contacts occurred within the first few weeks, but no late transmission took place. Public health interventions are needed to protect household contacts. PMID- 12533296 TI - Single multiplex polymerase chain reaction to detect diverse loci associated with diarrheagenic Escherichia coli. AB - We developed and tested a single multiplex polymerase chain reaction (PCR) that detects enterotoxigenic, enteropathogenic, enteroinvasive, and Shiga toxin producing Escherichia coli. This PCR is specific, sensitive, and rapid in detecting target isolates in stool and food. Because of its simplicity, economy, and efficiency, this protocol warrants further evaluation in large, prospective studies of polymicrobial substances. PMID- 12533297 TI - Risk for Pneumocystis carinii transmission among patients with pneumonia: a molecular epidemiology study. AB - We report a molecular typing and epidemiologic analysis of Pneumocystis carinii pneumonia (PCP) cases diagnosed in our geographic area from 1990 to 2000. Our analysis suggests that transmission from patients with active PCP to susceptible persons caused only a few, if any, PCP cases in our setting. PMID- 12533298 TI - First isolation of Rickettsia slovaca from a patient, France. PMID- 12533299 TI - Enteropathogenic Klebsiella pneumoniae HIV-infected adults, Africa. PMID- 12533300 TI - Granulomatous lymphadenitis as a manifestation of Q Fever. PMID- 12533301 TI - Has Coxiella burnetii (Q fever) been introduced into New Zealand? PMID- 12533302 TI - Effects of exogenous gibberellin and auxin on shoot elongation and vegetative bud development in seedlings of Pinus sylvestris and Picea glauca. AB - The hormonal control of stem unit (foliar appendage and axillary structure, if present, plus subtending internode) number and length was investigated in shoots of Scots pine (Pinus sylvestris L.) and white spruce (Picea glauca (Moench) Voss). Seedlings were treated with six gibberellins (GA1, GA3, GA4, GA5, GA9 and GA20) and two auxins (indole-3-acetic acid (IAA) and naphthaleneacetic acid (NAA)) when either neoformed growth was occurring or the terminal vegetative bud was developing. Hormones were applied by drenching the shoot tip, injecting the stem or spraying the foliage. Combined results for all three application methods indicated that shoot elongation in first-year seedlings (i.e., neoformed growth) was promoted in both species by GA1, GA3, GA4 and, less obviously, by GA9. This promotion was attributable to an increase in length, rather than number, of stem units. However, the number of stem units formed during terminal bud development, as reflected in the number of needles (white spruce) or cataphylls (Scots pine) present on the shoot resulting from the terminal bud, was stimulated by GA1, GA3 and GA4 in both species and by GA9 in Scots pine. The GA-induced increase in the number of preformed stem units was associated with increased bud width in white spruce and increased bud length and resulting shoot length in Scots pine. In contrast, application of IAA or NAA either did not affect or inhibited both neoformed growth and terminal bud stem unit number, depending on the application method and concentration. We conclude that, in the Pinaceae, (1) GA stimulates the activity of both the subapical meristem during neoformed growth and the apical meristem during vegetative bud development, and (2) the early non hydroxylation pathway, via GA9, is the major route of GA biosynthesis. The role of auxin in the control of stem unit number and length remains to be resolved. PMID- 12533304 TI - Contrasting effects of elevated carbon dioxide concentration and temperature on Rubisco activity, chlorophyll fluorescence, needle ultrastructure and secondary metabolites in conifer seedlings. AB - Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) Karst.) seedlings were grown for 50 days in growth chambers in an ambient or twice ambient carbon dioxide concentration ([CO2]) at a day/night temperature of 19/12 degrees C or 23/16 degrees C. Although elevated [CO2] (EC) had only slight effects on the growth parameters measured, elevated temperature (ET) increased above ground dry mass of both species. Among treatments, biomass accumulation of both species was greatest in the combined EC + ET treatment. The EC treatment induced thylakoid swelling and increased numbers of plastoglobuli observed in Scots pine needles. Although EC had little effect on Rubisco protein or N concentration of needles, ET had a large effect on N-containing compounds and enhanced N allocation from 1-year-old needles. Terpenoids were more responsive to EC and ET than total phenolics. Generally, terpene concentrations were reduced by EC and increased by ET. Increased terpenoid concentrations in response to ET might be associated with thermotolerance of photosynthesis. In Norway spruce, EC decreased total phenolic concentrations in needles, probably as a result of increased growth. We conclude that, in seedlings of these boreal species, the effects of elevated [CO2] on the studied parameters were small compared with the effects of elevated temperature. PMID- 12533303 TI - Biochemical acclimation patterns of Betula pendula and Pinus sylvestris seedlings to elevated carbon dioxide concentrations. AB - Acclimation of photosynthesis to increasing atmospheric carbon dioxide concentration ([CO2]; 350 to 2,000 micromol mol-1) was followed in silver birch (Betula pendula Roth.) and Scots pine (Pinus sylvestris L.) seedlings for two years. Chlorophyll fluorescence and concentrations of Rubisco, chlorophyll, total soluble protein and nitrogen were monitored together with steady-state gas exchange at three CO2 concentrations (ambient [CO2] (345 +/- 20 micromol mol-1), the growth [CO2] and 1950 +/- 55 micromol mol-1). Rubisco and chlorophyll concentrations decreased in birch and Scots pine with increasing growth [CO2]. A nonlinear response was recorded for Rubisco and chlorophyll concentrations in birch, which was correlated with a significant decrease in specific leaf area. Nitrogen concentration decreased in birch leaves, but was unchanged in Scots pine needles. The species differed substantially in their steady-state CO2 exchange response to increasing growth [CO2]. The principal effect in birch was a significant nonlinear decrease in the steady-state gas exchange rate at the ambient [CO2], whereas in Scots pine the main effect was a significant increase in the steady-state gas exchange rate at the growth [CO2]. PMID- 12533305 TI - Leaf senescence and late-season net photosynthesis of sun and shade leaves of overstory sweetgum (Liquidambar styraciflua) grown in elevated and ambient carbon dioxide concentrations. AB - We examined the effects of elevated CO2 concentration ([CO2]) on leaf demography, late-season photosynthesis and leaf N resorption of overstory sweetgum (Liquidambar styraciflua L.) trees in the Duke Forest Free Air CO2 Enrichment (FACE) experiment. Sun and shade leaves were subdivided into early leaves (formed in the overwintering bud) and late leaves (formed during the growing season). Overall, we found that leaf-level net photosynthetic rates were enhanced by atmospheric CO2 enrichment throughout the season until early November; however, sun leaves showed a greater response to atmospheric CO2 enrichment than shade leaves. Elevated [CO2] did not affect leaf longevity, emergence date or abscission date of sun leaves or shade leaves. Leaf number and leaf area per shoot were unaffected by CO2 treatment. A simple shoot photosynthesis model indicated that elevated [CO2] stimulated photosynthesis by 60% in sun shoots, but by only 3% in shade shoots. Whole-shoot photosynthetic rate was more than 12 times greater in sun shoots than in shade shoots. In senescent leaves, elevated [CO2] did not affect residual leaf nitrogen, and nitrogen resorption was largely unaffected by atmospheric CO2 enrichment, except for a small decrease in shade leaves. Overall, elevated [CO2] had little effect on the number of leaves per shoot at any time during the season and, therefore, did not change seasonal carbon gain by extending or shortening the growing season. Stimulation of carbon gain by atmospheric CO2 enrichment in sweetgum trees growing in the Duke Forest FACE experiment was the result of a strong stimulation of photosynthesis throughout the growing season. PMID- 12533306 TI - Moderate shade can increase net gas exchange and reduce photoinhibition in citrus leaves. AB - Daily variations in net gas exchange, chlorophyll a fluorescence and water relations of mature, sun-acclimated grapefruit (Citrus paradisi Macfady.) and orange (Citrus sinensis L. Osbeck) leaves were determined in tree canopies either shaded with 50% shade screens or left unshaded (sunlit). Mean daily maximum photosynthetic photon flux density (PPFD) under shade varied from 500 to 700 micromol m-2 s-1 and was sufficient to achieve maximum net CO2 assimilation rates (A CO2). Responses of grapefruit and orange leaves to shading were remarkably similar. At midday, on bright clear days, the temperatures of sunlit leaves were 2-6 degrees C above air temperature and 1-4 degrees C above the temperatures of shaded leaves. Although midday depressions of stomatal conductance (gs) and A CO2 were observed in both sunlit and shaded leaves, shaded leaves had lower leaf-to air vapor pressure differences (D) along with higher gs, A CO2 and leaf water-use efficiency than sunlit leaves. Estimated stomatal limitation to A CO2 was generally less than 25% and did not differ between shaded and sunlit leaves. Leaf intercellular CO2 partial pressure was not altered by shade treatment and did not change substantially with increasing D. Radiation and high temperature stress induced non-stomatal limitation to A CO2 in sunlit leaves was greater than 40%. Reversible photoinhibition of photosystem II efficiency was more pronounced in sunlit than in shaded leaves. Thus, non-stomatal factors play a major role in regulating A CO2 of citrus leaves during radiation and high temperature stress. PMID- 12533307 TI - Shoot growth and crown development: effect of crown position in three-dimensional simulations. AB - Trees have been increasingly considered as modular organisms, with individual shoots forming autonomous units that respond semi-independently to their surrounding environment. However, there is evidence for fairly strict hormonal control of tree crown development. Studies on the hydraulic architecture of trees suggest a closer functional connection between shoots and crown development than is postulated by the theory of branch autonomy. We studied how shoot growth pattern influences growth and crown architecture in young Scots pine trees simulated by the LIGNUM model assuming that (a) the growth of a shoot mainly depends on its light climate and (b) the growth of a shoot is influenced by its position within the crown. We determined shoot position within the crown based on a recently developed vigor index. The vigor index compares the relative axis cross-sectional area from the base of the tree to each shoot and gives a value of 1 to the pathway of the greatest cross-sectional area. All other shoots attain values between 0 and 1 depending on their cross-sectional areas and the cross sectional areas of the branches leading there from the main axis. The shoot light climate is characterized by annually intercepted photosynthetically active radiation. We compared the results from simulations (a) and (b) against an independent data set. The addition of a within-shoot position index (the vigor index) to our simulation (simulation b) resulted in a more realistic tree form than that obtained with simulation (a) alone. We discuss the functional significance of the results as well as the possibilities of using an index of shoot position in simulations of crown architecture. PMID- 12533308 TI - Photosynthesis-nitrogen relationships: interpretation of different patterns between Pseudotsuga menziesii and Populus x euroamericana in a mini-stand experiment. AB - We compared photosynthesis-nitrogen relationships of one broad-leaved (poplar; Populus x euroamericana (Dole) Guinier) and one conifer (Douglas-fir; Pseudotsuga menziesii (Mirb.) Franco) species. Plants were grown in large pots to allow free root development and were kept well watered. We determined effects of low, intermediate and high nitrogen supply rates on area-based leaf nitrogen (Na) and chlorophyll concentrations, leaf mass per area (LMA), light-saturated photosynthesis (Amax), maximum carboxylation (Vcmax) and electron transport rate (Jmax), photosynthetic nitrogen-use efficiency (PNUE), and proportions of leaf N in active Rubisco (PR), bioenergetic pools (PB) and the light-harvesting complex (PLH). Nitrogen supply significantly affected leaf Na. Leaf mass per area did not differ between species and was unaffected by the N treatments. In both species, there was a positive correlation between leaf Na and chlorophyll concentration, and between leaf Na and the photosynthetic parameters Amax, Jmax and Vcmax. At comparable leaf Na, however, poplar showed twofold higher PNUE and a threefold steeper slope of the Amax- nitrogen relationship than Douglas-fir. Leaf Na was negatively correlated with PNUE in Douglas-fir but not in poplar. Leaf Na was also negatively correlated with PR, PB and PLH in Douglas-fir, whereas in poplar, a negative correlation was found only for PLH. Parameter PR was significantly higher in poplar than in Douglas-fir. The ratio of CO2 concentration in the intercellular space to that in ambient air was higher in poplar than in Douglas fir. Overall, our data suggest that differences in the photosynthesis-nitrogen relationship and PNUE between Douglas-fir and poplar primarily reflect a different investment of N to active Rubisco, and possibly a different constraint to CO2 diffusion. PMID- 12533309 TI - Protein transport across the lung epithelial barrier. AB - Alveolar lining fluid normally contains proteins of important physiological, antioxidant, and mucosal defense functions [such as albumin, immunoglobulin G (IgG), secretory IgA, transferrin, and ceruloplasmin]. Because concentrations of plasma proteins in alveolar fluid can increase in injured lungs (such as with permeability edema and inflammation), understanding how alveolar epithelium handles protein transport is needed to develop therapeutic measures to restore alveolar homeostasis. This review provides an update on recent findings on protein transport across the alveolar epithelial barrier. The use of primary cultured rat alveolar epithelial cell monolayers (that exhibit phenotypic and morphological traits of in vivo alveolar epithelial type I cells) has shown that albumin and IgG are absorbed via saturable processes at rates greater than those predicted by passive diffusional mechanisms. In contrast, secretory component, the extracellular portion of the polymeric immunoglobulin receptor, is secreted into alveolar fluid. Transcytosis involving caveolae and clathrin-coated pits is likely the main route of alveolar epithelial protein transport, although relative contributions of these internalization steps to overall protein handling of alveolar epithelium remain to be determined. The specific pathways and regulatory mechanisms responsible for translocation of proteins across lung alveolar epithelium and regulation of the cognate receptors (e.g., 60-kDa albumin binding protein and IgG binding FcRn) expressed in alveolar epithelium need to be elucidated. PMID- 12533310 TI - Delta 9-tetrahydrocannabinol disrupts mitochondrial function and cell energetics. AB - We have observed rapid and extensive depletion of cellular energy stores by Delta(9)-tetrahydrocannabinol (THC) in the pulmonary transformed cell line A549. ATP levels declined dose dependently with an IC(50) of 7.5 microg/ml of THC after 24-h exposure. Cell death was observed only at concentrations >10 microg/ml. Studies using JC-1, a fluorescent probe for mitochondrial membrane potential, revealed diminished mitochondrial function at THC concentrations as low as 0.5 microg/ml. At concentrations of 2.5 or 10 microg/ml of THC, a decrease in mitochondrial membrane potential was observed as early as 1 h after THC exposure. Mitochondrial function remained diminished for at least 30 h after THC exposure. Flow cytometry studies on cells exposed to particulate smoke extracts indicate that JC-1 red fluorescence was fivefold lower in cells exposed to marijuana smoke extract relative to cells exposed to tobacco smoke extract. Comparison with a variety of mitochondrial inhibitors demonstrates that THC produced effects similar to that of carbonyl cyanide p-trifluoromethoxyphenylhydrazone, suggesting uncoupling of electron transport. Loss of red JC-1 fluorescence by THC was suppressed by cyclosporin A, suggesting mediation by the mitochondrial permeability transition pore. This disruption of mitochondrial function was sustained for at least 24 h after removal of THC by extensive washing. These results suggest that exposure of the bronchopulmonary epithelium to THC may have important health and physiological consequences. PMID- 12533311 TI - Peroxisome proliferator ameliorates endothelial dysfunction in a murine model of hyperhomocysteinemia. AB - To test the hypothesis that endothelial dysfunction in hyperhomocysteinemia was due to increased levels of nitrotyrosine and matrix metalloproteinase (MMP) activity in response to antagonism of peroxisome proliferator-activated receptor alpha (PPAR-alpha), cystathionine beta-synthase (CBS) -/+ mice were bred, tail tissue was analyzed for genotype by PCR, and tail vein blood was analyzed for homocysteine (Hcy) by spectrofluorometry. To induce PPAR-alpha, mice were administered 8 microg/ml of ciprofibrate (CF) and grouped: 1) wild type (WT), 2) WT + CF, 3) CBS, 4) CBS + CF (n = 6 in each group). In these four groups of mice, plasma Hcy was 3.0 +/- 0.2, 2.5 +/- 1.2, 15.2 +/- 2.6 (P < 0.05 compared with WT), 11.0 +/- 2.9 micromol/l. Mouse urinary protein was 110 +/- 11, 86 +/- 6, 179 +/- 13, 127 +/- 9 microg.day(-1). kg(-1) by Bio-Rad dye binding assay. Aortic nitrotyrosine was 0.099 +/- 0.012, 0.024 +/- 0.004, 0.132 +/- 0.024 (P < 0.01 compared with WT), 0.05 +/- 0.01 (scan unit) by Western analysis. MMP-2 activity was 0.053 +/- 0.010, 0.024 +/- 0.002, 0.039 +/- 0.009, 0.017 +/- 0.006 (scan unit) by zymography. MMP-9 was specifically induced in CBS -/+ mice and inhibited by CF treatment. Systolic blood pressure (SPB) was 90 +/- 2, 88 +/- 16, 104 +/- 8 (P < 0.05 compared with WT), 96 +/- 3 mmHg. Aortic wall stress [(SPB. radius(2)/wall thickness)/2(radius + wall thickness)] was 10.2 +/- 1.9, 9.7 +/- 0.2, 16.6 +/- 0.8 (P < 0.05 compared with WT), 13.1 +/- 2.1 dyn/cm(2). The results suggest that Hcy increased aortic wall stress by increasing nitrotyrosine and MMP-9 activity. PMID- 12533312 TI - FGF-10 prevents mechanical stretch-induced alveolar epithelial cell DNA damage via MAPK activation. AB - Cyclic stretch of alveolar epithelial cells (AEC) can alter normal lung barrier function. Fibroblast growth factor-10 (FGF-10), an alveolar type II cell mitogen that is critical for lung development, may have a role in promoting AEC repair. We studied whether cyclic stretch induces AEC DNA damage and whether FGF-10 would be protective. Cyclic stretch (30 min of 30% strain amplitude and 30 cycles/min) caused AEC DNA strand break formation, as assessed by alkaline unwinding technique and DNA nucleosomal fragmentation. Pretreatment of AEC with FGF-10 (10 ng/ml) blocked stretch-induced DNA strand break formation and DNA fragmentation. FGF-10 activated AEC mitogen-activated protein kinase (MAPK), and MAPK inhibitors prevented FGF-10-induced AEC MAPK activation and abolished the protective effects of FGF-10 against stretch-induced DNA damage. In addition, a Grb2-SOS inhibitor (SH(3)b-p peptide), a RAS inhibitor (farnesyl transferase inhibitor 277), and a RAF-1 inhibitor (forskolin) each prevented FGF-10-induced extracellular signal regulated kinase (ERK) 1/2 phosphorylation in AEC. Moreover, N17-A549 cells that express a RAS dominant/negative protein prevented the FGF-10-induced ERK1/2 phosphorylation and RAS activation in AEC. We conclude that cyclic stretch causes AEC DNA damage and that FGF-10 attenuates these effects by mechanisms involving MAPK activation via the Grb2-SOS/Ras/RAF-1/ERK1/2 pathway. PMID- 12533313 TI - Modulation of sodium transport in fetal alveolar epithelial cells by oxygen and corticosterone. AB - Regulation of active Na(+) transport across fetal distal lung epithelial cells (FDLE) by corticosterone (CST), corticotropin-releasing hormone (CRH), and oxygen tension may be crucial for postnatal adaptation. FDLE isolated from 19-day rat fetuses (term: 22 days) were grown on permeable supports to confluent monolayers (duration 3 days) in 2.5, 5, 12, or 20% O(2) with 5% CO(2)-balance N(2) and mounted in Ussing chambers for measurement of short-circuit currents (I(sc)). FDLE monolayers grown in 20% O(2) had significantly higher levels of total I(sc) and of their amiloride-sensitive (I(amil)) and ouabain-sensitive (I(ouab)) components than hypoxic cells. Values (microA/cm(2) +/- SE) for 2.5-5% O(2) and 20% O(2) were, respectively, I(sc) 5.3 +/- 0.2 vs. 8.4 +/- 0.3 (P < 0.001), I(amil) 3.4 +/- 0.2 vs. 4.3 +/- 0.2 (P < 0.01), and I(ouab) 3.4 +/- 0.6 vs. 9.1 +/- 0.6 (P < 0.001). Addition of CST but not CRH to the culture medium at any O(2) concentration increased I(amil). FDLE cells grown at 5% O(2) expressed significantly lower levels of alpha-, beta-, and gamma-epithelial Na(+) channel (ENaC), and of the alpha(1)-Na(+)-K(+)-ATPase, as determined by Western blotting. We conclude that higher O(2) concentrations increased total vectorial Na(+) transport, and the function of Na(+)-K(+)-ATPase and apical amiloride-sensitive Na(+) conductance, whereas CST only increased ENaC function. PMID- 12533314 TI - Mechanical ventilation-induced pneumoprotein CC-16 vascular transfer in rats: effect of KGF pretreatment. AB - After air-blood barrier injury, "pneumoproteins" specific to lung epithelial distal airspaces reaching the bloodstream are putative markers of lung hyperpermeability. The contribution of mechanical ventilation (MV) to this leakage is unknown. To explore this issue, 16-kDa Clara cell protein (CC-16) concentration was quantified in bronchoalveolar lavages (BALFs) and/or sera of rats first exposed either to ambient air or to 48 h of hyperoxia-induced acute lung injury and then ventilated for 2 h according to one of the following strategies: 1) spontaneous ventilation (SV), 2) very-low-volume high PEEP (VLVHP, where PEEP is positive end-expiratory pressure), 3) low-volume zero PEEP, 4) moderate-volume low PEEP, and 5) high-volume zero PEEP (HVZP). Results show that total proteins in BALFs increased with time and MV, with little impact from hyperoxia preexposure. CC-16 content decreased in BALFs but increased in the bloodstream during MV, suggesting intravascular leakage. Lung overdistension may result either from high-volume (HVZP) or high-PEEP (VLVHP) MV, and it was the most potent inducer of CC-16 leakage (P < 0.05 vs. SV). In the VLVHP group, pretreatment with keratinocyte growth factor was efficient in reducing blood CC 16 transfer. PMID- 12533315 TI - Retinoic acid receptor-alpha regulates pulmonary alveolus formation in mice after, but not during, perinatal period. AB - The formation of pulmonary alveoli in mice and rats by subdivision of alveolar saccules that constitute the newborn's gas-exchange region ends by approximately postnatal day 14. However, alveoli continue to form after age 14 days until age approximately 40 days by means other than septation of the saccules present at birth. With the use of morphometric procedures and retinoic acid receptor (RAR) alpha+/+ and RAR-alpha-/- mice, we now show the volume of individual alveoli (va), the number of alveoli (Na), and alveolar surface area (Sa) are the same in 14-day-old RAR-alpha+/+ and RAR-alpha-/- mice. However, at age 50 days, va is larger, and Na and Sa are smaller, in RAR-alpha-/- than in RAR-alpha+/+ mice, although total lung volume is the same in both groups. These findings, and prior data showing RAR-beta is an endogenous inhibitor of alveolus formation during, but not after, the perinatal period, indicate there are developmental period specific regulators of alveolus formation and that total lung volume and alveolar dimensions may have different regulators. PMID- 12533317 TI - Tetrahedral gradient diffusion-encoding scheme with line scan imaging. PMID- 12533318 TI - Language processing during natural sleep in a 6-year-old boy, as assessed with functional MR imaging. PMID- 12533319 TI - Line scan diffusion imaging of the spine. AB - BACKGROUND AND PURPOSE: Recent findings suggest that diffusion-weighted imaging might be an important adjunct to the diagnostic workup of disease processes in the spine, but physiological motion and the challenging magnetic environment make it difficult to perform reliable quantitative diffusion measurements. Multi section line scan diffusion imaging of the spine was implemented and evaluated to provide quantitative diffusion measurements of vertebral bodies and intervertebral disks. METHODS: Line scan diffusion imaging of 12 healthy study participants and three patients with benign vertebral compression fractures was performed to assess the potential of line scan diffusion imaging of the spinal column. In a subgroup of six participants, multiple b-value (5-3005 s/mm(2)) images were obtained to test for multi-exponential signal decay. RESULTS: All images were diagnostic and of high quality. Mean diffusion values were (230 +/- 83) x 10(-6) mm(2)/s in the vertebral bodies, (1645 +/- 213) x 10(-6) mm(2)/s in the nuclei pulposi, (837 +/- 318) x 10(-6) mm(2)/s in the annuli fibrosi and ranged from 1019 x 10(-6) mm(2)/s to 1972 x 10(-6) mm(2)/s in benign compression fractures. The mean relative intra-participant variation of mean diffusivity among different vertebral segments (T10-L5) was 2.97%, whereas the relative difference in mean diffusivity among participants was 7.41% (P <.0001). The estimated measurement precision was <2%. A bi-exponential diffusion attenuation was found only in vertebral bodies. CONCLUSION: Line scan diffusion imaging is a robust and reliable method for imaging the spinal column. It does not suffer as strongly from susceptibility artifacts as does echo-planar imaging and is less susceptible to patient motion than are other multi-shot techniques. The different contributions from the water and fat fractions need to be considered in diffusion weighted imaging of the vertebral bodies. PMID- 12533320 TI - Anterior spinal artery and artery of Adamkiewicz detected by using multi-detector row CT. AB - BACKGROUND AND PURPOSE: Our purpose was to evaluate the visualization of the artery of Adamkiewicz (AKA) and the anterior spinal artery (ASA) by using multi detector row CT. Preoperative detection of the AKA and ASA is important for prevention of ischemic complications of thoracoabdominal aortic surgery. METHODS: Data from contrast-enhanced CT of the abdomen of 19 consecutive patients with known or suspected liver disease were evaluated. The scanning range was set from the level of the diaphragm to the lower edge of the liver. After bolus injection of contrast material (100 mL of iohexol, 350 mgI/mL, 5 mL/s), arterial phase scans were obtained by using a four-channel multi-detector row CT scanner. The scanning parameters included a detector row configuration of 4 x 2 mm, a pitch of 5:1, a gantry rotation speed of 0.5 s, 120 kVp, and 150 mAs. Arterial phase coronal multiplanar reconstruction scans obtained parallel to the spinal cord were evaluated by two neuroradiologists. The detectability of ASA and AKA was analyzed. RESULTS: The AKA was visualized on the scans of 13 of 19 patients (68%). The segmental level of AKA origin ranged from T10 to L2. The AKA originated from the left side in nine patients (69%) and the right side in four patients (31%). The ASA was visualized on the scans of all patients (100%). For 16 of the 19 patients, the ASA was detected in its full length from the cranial edge of the scan range. However, the ascending branch of the ASA distal to the junction of the AKA was not detected for the remaining three patients. CONCLUSION: The AKA and ASA can be visualized by using multi-detector row CT with the use of IV administered contrast material. Multi-detector row CT could be a useful tool in the evaluation of spinal vascular structures. PMID- 12533321 TI - MR Imaging in Wegener granulomatosis of the spinal cord. AB - Wegener granulomatosis is a multisystemic disorder characterized by necrotizing vasculitis that primarily involves the respiratory tract. The orbits, heart, skin, joints, and nervous system are frequently involved. We describe the MR imaging findings of Wegener granulomatosis in the cervical spine and correlate them with the histopathologic features. MR imaging showed epidural liquid masses surrounded by granulomatous masses that compressed the cervical myelin from behind. PMID- 12533322 TI - Postmortem MR imaging of the fetal and stillborn central nervous system. AB - BACKGROUND AND PURPOSE: Changes in the public perception of postmortem procedures in the United Kingdom have led to reduced numbers of autopsies being performed in the fetus and neonate. When autopsy is performed in this group, the brain is now usually studied without being formalin-fixed, which limits the available information. We evaluated the diagnostic accuracy of postmortem MR imaging of the fetal brain and spine when compared with the reference standard, autopsy. METHODS: We obtained high-spatial-resolution T2-weighted images (in-plane resolution approximately 0.4 mm) of the brain and spine in 40 fetuses and stillborn neonates (14-42 weeks gestational age) who were referred for autopsy. The MR findings were compared with those of autopsy, the reference standard, which had been performed independently. RESULTS: In eight cases, the autopsy did not provide structural information of the brain or spine, because assessment of the unfixed tissue was impossible. There was agreement between MR and autopsy findings in 31 (97%) of 32 cases in which comparison could be made. Eleven cases showed normal brain, and 20 cases showed a wide range of developmental and acquired abnormalities. The sensitivity of MR was 100%, specificity 92%, positive predictive value 95%, and negative predictive value 100%. CONCLUSION: MR imaging has a useful role in providing structural information of the central nervous system in fetuses and stillborn neonates. PMID- 12533323 TI - Fetal cytomegalovirus infection of the brain: the spectrum of sonographic findings. AB - BACKGROUND AND PURPOSE: Congenital cytomegalovirus (CMV) infection can cause a wide range of brain anomalies. These changes have been well described postnatally, but descriptions of their in utero evolution are scarce. The purpose of this study was to analyze the sonographic spectrum of intracranial abnormalities in fetuses with proved CMV infection and to determine characteristic patterns of this infection. METHODS: We reviewed the transabdominal and transvaginal sonograms of eight fetuses with proved CMV infection. The sonographic analysis searched for signs of ventriculitis, leukomalacia, calcification, vasculitis, and periventricular cyst formation. The gyral pattern, corpus callosum, and cerebellar morphology also were examined. Fetal MR imaging was performed in two cases. RESULTS: The sonographic diagnosis of suspected CMV infection was made at a mean gestational age of 27.5 weeks (range, 22-37 weeks). An abnormal pattern of periventricular echogenicity was found in all fetuses. Echogenic intraparenchymal foci and ventriculomegaly were present in five fetuses. Other signs of fetal infection were intraventricular adhesions, periventricular pseudocysts, sulcation and gyral abnormal patterns, hypoplastic corpus callosum, cerebellar and cisterna magna abnormalities, and signs of striatal artery vasculopathy. For all fetuses, transvaginal sonography provided additional information. Fetal MR imaging provided additional information in one case. CONCLUSION: The presence of the described sonographic findings, particularly if two or more are present in the same fetus, is an indication for CMV investigation. PMID- 12533324 TI - Proton MR spectroscopy in the diagnostic evaluation of suspected mitochondrial disease. AB - PURPOSE AND BACKGROUND: Mitochondrial diseases are a group of inherited disorders caused by a derangement of mitochondrial respiration. The clinical manifestations are heterogeneous, and the diagnosis is often based on information acquired from multiple levels of inquiry. MR spectroscopy has previously been shown to help detect an abnormal accumulation of lactate in brain parenchyma and CSF in association with mitochondrial disorders, but the frequency of detection is largely unknown. We sought to examine the frequency of detectable elevations of CNS lactate by proton MR spectroscopy in a population of children and young adults with suspected mitochondrial disease. METHODS: MR spectroscopy data evaluated for the presence or absence of abnormal brain or CSF lactate were compared with other clinical indicators of mitochondrial dysfunction for 29 patients with suspected mitochondrial disease during the years 1990 to 2000. Based on an independent review of the final diagnoses, the patients were divided into groups based on the probability of mitochondrial disorder. RESULTS: A total of 32 scans from 29 patients were reviewed. Of eight patients thought to have a definitive mitochondrial disorder on the basis of genetic, biochemical, or pathologic features, five were found to have abnormal brain or CSF lactate levels revealed by MR spectroscopy (for one patient in whom two images were acquired, one was negative and the other positive). Among the studies conducted using a multisection spectroscopic imaging technique, five of six showed elevated lactate in the brain parenchyma, six of six showed elevated lactate in the CSF, and five of six showed elevated lactate in both brain and CSF. Of 16 patients who were highly suspected of having mitochondrial disorders on the basis of clinical grounds alone but who were lacking genetic, biochemical, or pathologic confirmation, four had abnormal lactate levels shown by MR spectroscopy. Mitochondrial disorder was excluded for five patients, none of whom had CNS lactate shown by MR spectroscopy. CONCLUSION: Detection of CNS lactate by MR spectroscopy is useful in the diagnosis of mitochondrial disease. In our series of patients with confirmed mitochondrial disease, a high level of lactate shown by MR spectroscopy correlated well with other markers of mitochondrial disease. As with all other means used to diagnose mitochondrial disorders, MR spectroscopy does not depict elevated lactate in all cases. Abnormal CNS concentrations of lactate may be undetected by MR spectroscopy because of differences in the type of mitochondrial disorder, timing, severity, or location of the affected tissues and the site of interrogation. PMID- 12533325 TI - Language processing during natural sleep in a 6-year-old boy, as assessed with functional MR imaging. AB - Functional MR imaging for language lateralization was performed in a 6-year-old child before neurosurgical intervention. A passive story-listening task was used; this revealed a bilateral language network. The task was repeated during the same session when the child had fallen asleep and surprisingly yielded strong activation in similar language areas. Our findings suggest that language processing does occur during natural sleep, even in young children. This potentially allows for an assessment of language functions, even in sleeping children. PMID- 12533326 TI - The jugular foramen in complex and syndromic craniosynostosis and its relationship to raised intracranial pressure. AB - BACKGROUND AND PURPOSE: Complex and syndromic craniosynostosis can be complicated by raised intracranial pressure (ICP), which in the absence of other identifiable origins, is probably caused by venous hypertension. Children with these conditions have been shown to have narrowing of the sigmoid sinus-jugular vein complex. Evidence of bony narrowing of the jugular foramina in children with complex or syndromic craniosynostosis and raised ICP compared with that in children with craniosynostosis without raised ICP would provide support for the theory that venous hypertension occurs in the former children. METHODS: Measurements of the jugular foramina were obtained from reformatted helical CT scans obtained in 12 children with complex or syndromic craniosynostosis and raised ICP (group 1) and in two control groups of children with normal ICP. The first control group comprised 10 children with simple nonsyndromic synostosis of one or two sutures (group 2), and the second control group included nine children with complex or syndromic craniosynostosis (group 3). RESULTS: Children with raised ICP had narrower jugular foramina than did the age-matched control subjects. For group 1, the mean diameter of jugular foramina was 6.5 mm; group 2, 11.5 mm (P <.01); and group 3, 10 mm (P <.05). No significant difference existed between the two control groups. CONCLUSION: Significantly narrower jugular foramina in children with raised ICP is further evidence of the role of venous outflow obstruction and intracranial venous hypertension in the development of raised ICP in complex and syndromic craniosynostosis. PMID- 12533327 TI - Increased diffusion in the brain of professional boxers: a preclinical sign of traumatic brain injury? AB - BACKGROUND AND PURPOSE: Professional boxing is associated with chronic, repetitive head blows that may cause brain injuries. Diffusion-weighted imaging is sensitive to microscopic changes and may be a useful tool to quantify the microstructural integrity of the brain. In this study, we sought to quantify microscopic alterations associated with chronic traumatic brain injury in professional boxers. METHODS: MR and diffusion-weighted imaging were performed in 24 boxers and in 14 age- and sex-matched control subjects with no history of head trauma. Using distribution analysis, the average diffusion constant of the entire brain (BD(av)) and diffusion distribution width (sigma) were calculated for each subject; findings in professional boxers were compared with those of control subjects. In the boxer group, correlations between diffusion changes and boxing history and diffusion changes and MR imaging findings were assessed. RESULTS: The measured diffusion values in the boxer group were significantly higher than those measured in the control group (BD(av), P <.0001; sigma, P <.01). In the boxer group, a robust correlation was found between increased BD(av) and frequency of hospitalization for boxing injuries (r = 0.654, P <.05). The most common MR finding in the boxer group was volume loss inappropriate to age followed by cavum septum pellucidum, subcortical white matter disease, and periventricular white matter disease. CONCLUSION: Boxers had higher diffusion constants than those in control subjects. Our data suggest that microstructural damage of the brain associated with chronic traumatic brain injury may elevate whole-brain diffusion. This global elevation can exist even when routine MR findings are normal. PMID- 12533328 TI - Standardized, reproducible, high resolution global measurements of T1 relaxation metrics in cases of multiple sclerosis. AB - BACKGROUND AND PURPOSE: We herein present a methodology for standardized and clinically applicable measurement of T1 relaxation maps with high resolution and volumetric coverage by using the commercially available 3D spoiled gradient-echo sequence. The reproducibility of the T1 metrics derived from these maps and their sensitivity to distinguish between control participants and patients with multiple sclerosis are evaluated. METHODS: Axial view 3D RF spoiled data sets with two flip angles were acquired at 1.5 T to generate the T1 maps, with all other imaging parameters (27/6 ms [TR/TE]; field of view, 180 x 240 x 186 mm(3); matrix, 192 x 256 x 124) kept identical between the two acquisitions. T1 maps were collected from 20 normal control participants and 32 patients with multiple sclerosis. An automated and operator-independent method was developed to segment the relaxation maps and define T1 metrics. RESULTS: We showed that the metrics derived from these maps to represent tissue characteristics were highly reproducible (coefficient of variation, approximately 1% to 4%) and were significantly different between normal control participants and patients with multiple sclerosis (P <.001) for the small cohort of patients in this study. CONCLUSION: The commercially accessible 3D spoiled gradient-echo sequence can be used to generate T1 relaxation maps with high resolution and volumetric coverage. The metrics derived from the relaxation maps are reproducible and have been shown to be sensitive to qualitative and quantitative differences between subgroups of patients with multiple sclerosis and control participants, with strong statistical significance. The use of a commercially available sequence enables the standardization and comparison of T1 metrics across different multiple sclerosis centers. PMID- 12533329 TI - Brain metabolite profiles of T1-hypointense lesions in relapsing-remitting multiple sclerosis. AB - BACKGROUND AND PURPOSE: Persistent T1-hypointense lesions ("black holes") are thought to represent permanent damage of brain parenchyma. We attempted to ascertain whether the metabolic profiles of these hypointense areas support this hypothesis and whether these profiles correlate with these hypointense findings. METHODS: Four patients with relapsing-remitting multiple sclerosis and four matched control volunteers underwent MR imaging and 3D proton MR spectroscopy. Absolute levels of N-acetylaspartate (NAA), creatine, and choline (Cho) were obtained in 0.19 cm(3) voxels containing 14 T1-hypointense lesions (average volume, 0.4 cm(3); range, 0.2-1.0 cm(3)) in patients. Metabolite levels were analyzed, by using Pearson correlation, against their respective lesions' hypointensity relative to the surrounding normal-appearing white matter. RESULTS: Moderate correlation, r = 0.56, was found between the NAA level and MR imaging hypointensity. Of the 14 lesions studied, 12 were deficient in NAA and 11 had excess Cho compared with corresponding brain regions in control volunteers. Only one lesion was significantly deficient in all three metabolites, indicative of total damage or matrix loss. CONCLUSION: No relationship was found between the hypointensity of the lesions and their metabolic profile. Specifically, lesions with the same hypointensity on T1-weighted MR images were metabolically variable (ie, displayed disparate metabolite levels and behavior). Also, although 86% of the lesions exhibited abnormally low NAA, 71% also had increased Cho. This indicates that although neuronal damage had already occurred (lower NAA), these lesions were still "smoldering" with active membrane turnover (high Cho), most likely because of de- and remyelination, indicative of shadow plaques (remyelinated lesions). Consequently, relapsing-remitting hypointense lesions represent neither final-stage nor static pathologic abnormality. PMID- 12533330 TI - Short-term correlations between clinical and MR imaging findings in relapsing remitting multiple sclerosis. AB - BACKGROUND AND PURPOSE: Despite extensive use of MR imaging to provide markers of multiple sclerosis (MS) activity and accumulated disease burden, the magnitude of the relationship between clinical and MR findings is still debated. Using data from the European/Canadian glatiramer acetate (GA) trial, we investigated short term correlations between clinical and MR measures of disease activity in patients with relapsing-remitting MS (RRMS). METHODS: In a 9-month, double blinded, placebo-controlled study, 239 patients with RRMS were randomly assigned to receive 20 mg GA (n = 119) or placebo (n = 120). Clinical assessment included monthly neurologic examinations with Expanded Disability Status Scale scoring and visits for symptoms suggestive of relapse. Dual-echo T2-weighted and pre- and postcontrast T1-weighted brain MR images were obtained at baseline and monthly during follow-up. Contrast-enhancing and new T2-hyperintense lesions were counted, and total T2-hyperintense and T1-hypointense lesion volumes were measured. RESULTS: Significant univariate correlations were found between the number of relapses during the study period and the number of enhancing lesions at baseline (r = 0.25) and during follow-up (r = 0.30) in the study population as a whole. Multivariable analysis showed that two independent factors were more strongly correlated with relapse frequency: the number of relapses during the 2 years before entry and the number of on-trial enhancing lesions, in the whole study population and in the placebo group. CONCLUSION: In RRMS, MR imaging measures of inflammatory activity are modestly but significantly correlated with the occurrence of clinical attacks over the short term. Clinical and MR imaging assessment can provide complementary outcome measures for RRMS trials. PMID- 12533331 TI - Whole brain imaging of HIV-infected patients: quantitative analysis of magnetization transfer ratio histogram and fractional brain volume. AB - BACKGROUND AND PURPOSE: Magnetization transfer ratio (MTR) histogram analysis and volumetric MR imaging are sensitive tools with which to quantify the tissue destructive effects in patients with white matter or neurodegenerative disease. Our purpose was to determine whether whole brain MTR and fractional brain parenchyma volume measurements are altered in HIV-1-infected patients who are neurologically symptomatic and in those who are asymptomatic. METHODS: We performed MR imaging and MTR studies of 15 neurologically symptomatic (seven patients) and asymptomatic (eight patients) HIV-1-seropositive patients and compared their findings with those of 10 seronegative normal control participants. MTR was computed on the basis of whole brain parenchyma segmented by using thin section dual echo MR images. RESULTS: The loss of brain tissue, indicated by fractional brain parenchyma volume, was more pronounced in neurologically symptomatic patients (P =.003) but not in asymptomatic patients (P =.23) when compared with control participants. As for whole brain MTR histogram analysis, both patient groups showed significant decrease in mean (P =.02) and median (P < or =.009) values, compared with normal control participants. There was a trend toward positive correlation (r > or = 0.56) between MTR histogram statistics and fractional brain parenchyma volume. CONCLUSION: Our results suggest that MTR histogram analysis is sensitive in detecting early involvement in neurologically asymptomatic patients with HIV and may, therefore, be used as a combined tool with volumetric measurement, which showed significant tissue loss only in symptomatic patients, to assess various stages of brain damage induced by HIV. PMID- 12533332 TI - Brain microhemorrhages detected on T2*-weighted gradient-echo MR images. AB - BACKGROUND AND PURPOSE: Multifocal microhemorrhages have been reported to be commonly found in the brain of patients with systemic hypertension and spontaneous brain hemorrhage. The factors associated with these lesions detected on T2*-weighted gradient-echo images were examined to determine whether these lesions serve to indicate different types of microangiopathy and to predict a patient's risk for symptomatic hemorrhage. METHODS: The study population consisted of 2164 patients who underwent 2416 consecutive brain MR imaging studies performed during 3 years. The patients with intracerebral hemorrhages due to vascular malformations, neoplasms, trauma, or intracranial surgery and those with incomplete medical records were excluded; 2019 cases were analyzed. RESULTS: The overall incidence of microhemorrhages was 9.8%, predominantly in the lentiform nucleus (n = 96), thalamus (n = 88), and cortical-subcortical region (n = 93). Presence of microhemorrhages had the highest significant correlation with history of hemorrhagic stroke (P <.0001); advancing age, hypertension, and prominent white matter hyperintensity on T2-weighted images had the next highest significant correlation. Cortical-subcortical microhemorrhages were more frequently observed in patients who had previous lobar hemorrhagic stroke (P <.005). Among 139 patients with microhemorrhages who could be clinically followed up for more than 1 month, four (2.9%) had new hemorrhagic stroke. CONCLUSION: The presence of microhemorrhages may be not only a direct marker of bleeding-prone small-vessel diseases but also an indicator of different types of microangiopathy and a predictor of further hemorrhagic stroke. PMID- 12533333 TI - Contrast-enhanced MR imaging of cerebral fat embolism: case report and review of the literature. AB - We report a case of cerebral fat embolism (CFE) that was imaged with T2- and T2* weighted, fluid-attenuated inversion recovery, and contrast-enhanced T1-weighted images. Multiple white matter lesions demonstrated contrast enhancement, with little evidence of hemorrhage. The patient regained complete neurologic function. CFE can cause capillary inflammation and breakdown of the blood-brain barrier, without associated hemorrhage or permanent neurologic deficit. In this case, both contrast-enhanced T1-weighted and gradient-echo T2*-weighted images were important in diagnosing CFE and in determining the severity of the patient's condition. PMID- 12533334 TI - Prolonged reversible vasospasm in cyclosporin A-induced encephalopathy. AB - The mechanisms responsible for cyclosporin-induced encephalopathy remain controversial. Herein we present a case of cyclosporin-induced encephalopathy with unusually prolonged vasospasm, which might have contributed to the slow recovery of the patient. PMID- 12533335 TI - Xanthoma disseminatum: a rare intracranial mass. AB - We report the CT, MR imaging, and histologic findings in a patient with an intracranial mass diagnosed as xanthoma disseminatum (XD). The mass demonstrated radiographic findings typical for a glioma. However, histologic examination of brain and dermal lesions showed findings characteristic of XD. XD is a rare dermatologic disorder of unknown cause that may be accompanied by systemic manifestations. In rare cases, it can appear as an intracranial mass. PMID- 12533336 TI - Imaging findings associated with childhood primary intracranial squamous cell carcinoma. AB - A 5-year-old girl with no preexisting systemic or CNS neoplasm presented with a large right temporal mass lesion, the histopathology of which proved to be a poorly differentiated adenosquamous carcinoma, a highly unusual primary intracranial tumor. The tumor recurred despite radical resection, chemotherapy, and radiation therapy. PMID- 12533337 TI - Unusual appearance of a cisternal hydatid cyst. AB - We herein describe a case of an isolated multilocular hydatid cyst occurring in the basilar cisterns, with nondependent levels of different signal intensities shown on MR images. Three portions were visualized on T1-weighted images: the hyperintense inferior aspect, the isointense central part, and the superior aspect of lower signal intensity. The inferior portion was hypointense on T2 weighted images. This unusual appearance was caused by layering of hydatid sand and may represent a characteristic feature of hydatid disease. PMID- 12533338 TI - Primary diffuse meningeal melanomatosis: radiologic-pathologic correlation. AB - We report a case of primary diffuse meningeal melanomatosis, a rare variant of primary malignant melanoma of the CNS, in a 68-year-old woman. The disease mimicked intracranial hypotension syndrome and was diagnosed only at autopsy (CSF cytologic results were negative). CT revealed hydrocephalus with effacement of the cerebral convexity sulci and abnormal contrast enhancement in the right sylvian and frontoparietal fissures, whereas MR imaging showed diffuse marked dural and leptomeningeal contrast enhancement. In retrospect, these nonspecific findings correlated with the extensive leptomeningeal invasion in the cerebral hemispheres, brain stem and spinal cord. The clinical, radiologic, and pathologic features of diffuse meningeal melanomatosis are reviewed. PMID- 12533339 TI - Idiopathic hypertrophic cranial pachymeningitis: case report with 7 years of imaging follow-up. AB - Idiopathic hypertrophic cranial pachymeningitis is a rare inflammatory disease with diffused involvement of the dura. Often, the definite diagnosis is made immediately with biopsy and the involved dura is removed surgically. Consequently, extensive preoperative imaging studies usually are not available. We reviewed a case of idiopathic hypertrophic cranial pachymeningitis and collectively summarized the interesting features from the 7 years preceding surgical treatment. These chronologic imaging findings with progressive intracranial involvement included dural thickening, dural mass, sinus thrombosis, and venous congestion constituted comprehensive pictures of idiopathic hypertrophic cranial pachymeningitis. The thickened dura may also at times mimic dural masses, such as en plaque meningioma. PMID- 12533340 TI - Angiographic documentation of a persistent otic artery. AB - The persistent otic artery is a rare remnant of early fetal development that, to our knowledge, has been convincingly documented only once before with angiography. Other persistent vessels such as the trigeminal, hypoglossal, and proatlantal intersegmental arteries have been found to be associated with a variety of vascular anomalies. We believe we present the second case of a persistent otic artery documented by means of angiography and the first such case associated with multiple intracranial aneurysms. PMID- 12533341 TI - Thromboembolic events associated with Guglielmi detachable coil embolization of asymptomatic cerebral aneurysms: evaluation of 66 consecutive cases with use of diffusion-weighted MR imaging. AB - BACKGROUND AND PURPOSE: Although Guglielmi detachable coil (GDC) endovascular treatment of intracranial aneurysms has become an accepted alternative to surgery, the main complication continues to be thromboembolic events. We sought to determine the frequency and radiologic appearance of thromboembolic events during GDC embolization for asymptomatic cerebral aneurysms by using diffusion weighted (DW) MR imaging and to determine whether aneurysmal anatomic factors or use of the balloon-assisted technique affected the frequency. METHODS: In 74 patients, 79 asymptomatic cerebral aneurysms were treated with GDC embolizations at the National Cardiovascular Center from 1999 to 2001. Thirty-nine of these aneurysms (49%) were treated with the balloon-assisted technique. DW imaging was performed in 66 patients at 2-5 days after GDC embolization. All DW images were reviewed by two radiologists for depiction of abnormalities. RESULTS: DW images showed hyperintense lesions in 40 patients (61%), with 16 of these patients (40%) incurring neurologic deteriorations. Fifteen of the symptomatic patients (94%) fully recovered by discharge, and the remaining one experienced permanent deficits. Hyperintense lesions were detected more frequently in wide-neck (73%) or large (100%) aneurysms and in procedures that used the balloon-assisted technique (73%) than in small aneurysms (50%) or in procedures with the simple GDC method (49%). The occurrence of new lesions was significantly associated with use of the balloon-assisted technique and with aneurysm diameter in multivariate analysis (P <.05). CONCLUSION: In our experience, thromboembolic events related to the use of GDC embolization are relatively common, especially in wide-neck or large aneurysms or in association with the balloon-assisted technique. Although permanent deficits are rare, the high rate of thromboembolic events suggests that improvements in the technique such as the addition of antiplatelet agents and the development of new embolic materials are mandatory. PMID- 12533342 TI - Combined endovascular treatment for both intracranial aneurysm and symptomatic vasospasm. AB - BACKGROUND AND PURPOSE: The best strategy for treatment of subarachnoid hemorrhage due to ruptured cerebral aneurysm is obliteration of the aneurysm as soon as possible. Early surgery is desirable if the patient does not develop severe vasospasm or is clinically stable. However, if the patient has already developed severe vasospasm on admission, surgery may carry the risk of increasing the severity. We evaluated the safety and effectiveness of combined Guglielmi detachable coil (GDC) embolization and angioplasty in a single session for the treatment of ruptured aneurysms associated with symptomatic vasospasm. METHODS: From January 1992 to January 2001, 12 consecutive patients with ruptured aneurysms associated with symptomatic vasospasm were treated. Patients were classified as Hunt and Hess grade 2 (n = 1), 3 (n = 6), 4 (n = 4), or 5 (n = 1) and Fisher CT group 2 (n = 1), 3 (n = 10), or 4 (n = 1). They underwent GDC aneurysm occlusion and balloon angioplasty (n = 6), intraarterial papaverine infusion (n = 2), or both (n = 4) in a single session. In nine patients, aneurysm coil occlusion was performed first. RESULTS: Complete GDC occlusion was achieved in eight patients, a small neck remnant persisted in three, and embolization was incomplete in one patient. In all patients, angiographic improvement of vasospasm was obtained. In one patient, a thromboembolic complication occurred and was treated with urokinase. Clinical outcomes at discharge were good recovery in six, moderate disability in two, severe disability in three, or death in one. CONCLUSION: Endovascular treatment can be the first therapeutic option for ruptured aneurysms associated with severe vasospasm on admission. It offers some advantages over surgery in this setting, but these are balanced by the risk of thromboembolism. PMID- 12533343 TI - CT angiography before embolization for hemorrhage in head and neck cancer. AB - We present a patient with advanced head and neck carcinoma and a bleeding pseudoaneurysm diagnosed by means of CT angiography; this was not apparent on conventional digital subtraction angiograms. The information provided by CT angiography facilitated rapid identification of the pseudoaneurysm and treatment with embolization. CT angiography may be helpful before embolization in cases of hemorrhage in head and neck cancer. PMID- 12533344 TI - CT and MR imaging findings associated with subacute thyroiditis. AB - Subacute granulomatous thyroiditis is an uncommon disease that occurs most often in women in their second to fifth decades of life. This disease usually presents with thyroid tenderness, a low grade fever, and occasional dysphagia. The disease resolves spontaneously, usually without thyroid function abnormalities. We herein present the CT and MR imaging findings of two cases of subacute granulomatous thyroiditis. PMID- 12533345 TI - Superficial temporal artery pseudoaneurysm: diagnosis and preoperative planning with CT angiography. AB - We report the use of CT angiography in the diagnosis and preoperative planning of a superficial temporal artery pseudoaneurysm. A 50-year-old man presented with a pulsatile preauricular mass 4 weeks after undergoing pterional craniotomy for aneurysm repair. CT angiography revealed a 2.5-cm pseudoaneurysm arising from the posterior margin of the right superficial temporal artery at the inferior margin of the craniotomy incision. To our knowledge, this is the first reported case showing the usefulness of CT angiography alone in the diagnosis and characterization of a superficial temporal artery pseudoaneurysm. PMID- 12533346 TI - Symptomatic enlarged cervical anterior epidural venous plexus in a patient with Marfan syndrome. PMID- 12533347 TI - Symptomatic enlarged cervical anterior epidural venous plexus in a patient with Marfan syndrome. PMID- 12533349 TI - Analysis and classification of cerebellar malformations. PMID- 12533352 TI - Detection and characterization of very small cerebral aneurysms by using 2D and 3D helical CT angiography. PMID- 12533353 TI - Multidetector-row CT angiographic findings in the bilateral aortic arch origin of the vertebral arteries. PMID- 12533357 TI - Rapid sequence intubation in Scottish urban emergency departments. AB - OBJECTIVE: Airway care is the cornerstone of resuscitation. In UK emergency department practice, this care is provided by anaesthetists and emergency physicians. The aim of this study was to determine current practice for rapid sequence intubation (RSI) in a sample of emergency departments in Scotland. METHODS: Two year, multicentre, prospective observational study of endotracheal intubation in the emergency departments of seven Scottish urban teaching hospitals. RESULTS: 1631 patients underwent an intubation attempt in the emergency department and 735 patients satisfied the criteria for RSI. Emergency physicians intubated 377 patients and anaesthetists intubated 355 patients. There was no difference in median age between the groups but there was a significantly greater proportion of men (73.2% versus 65.3%, p=0.024) and trauma patients (48.5% versus 37.4%, p=0.003) in the anaesthetic group. Anaesthetists had a higher initial success rate (91.8% versus 83.8%, p=0.001) and achieved more good (Cormack-Lehane Grade I and II) views at laryngoscopy (94.0% versus 89.3%, p=0.039). There was a non-significant trend to more complications in the group of patients intubated by emergency physicians (8.7% versus 12.7%, p=0.104). Emergency physicians intubated a higher proportion of patients with physiological compromise (91.8% versus 86.1%, p=0.027) and a higher proportion of patients within 15 minutes of arrival (32.6% versus 11.3%, p<0.0001). CONCLUSION: Anaesthetists achieve more good views at laryngoscopy with higher initial success rates during RSI. Emergency physicians perform RSI on a higher proportion of critically ill patients and a higher proportion of patients within 15 minutes of arrival. Complications may be fewer in the anaesthetists' group, but this could be related to differences in patient populations. Training issues for RSI and emergency airway care are discussed. Complication rates for both groups are in keeping with previous studies. PMID- 12533358 TI - Resuscitation Predictor Scoring Scale for inhospital cardiac arrests. AB - OBJECTIVES: The purpose of this study was to determine the key factors influencing survival from cardiopulmonary resuscitation attempts and to produce a survival predictor scale for use during a resuscitation attempt. METHOD: Bivariate analysis of individual survival predictors and a prospective analysis of survival based on logistic regression models. Included in this seven year study (1993-2000) were 2567 inhospital resuscitation calls of which 1633 received full cardiopulmonary resuscitation. Immediate, 24 hour and discharge survival rates were the main outcome measures with additional analysis for the development of the Resuscitation Predictor Scoring Scale (RPS Scale). RESULTS: The immediate survival rate was 41%, 28% at 24 hours, and 19% by discharge. Multivariate analysis showed the main factors influencing 24 hour survival to be the duration of the arrest, primary arrhythmia (VT, VF, asystole, or PEA), age, and the primary mode of arrest (respiratory or cardiac). The RPS Scale was developed from these key predictors giving resuscitation teams an accurate prediction of survival 15 minutes into a resuscitation attempt. CONCLUSION: Data collection and analysis of cardiopulmonary resuscitation attempts are essential for the formulation of survival indicators. In this case the data have enabled the formulation of a survival predictor scale that will quantify the decision making process regarding the termination of cardiopulmonary resuscitation attempts. PMID- 12533359 TI - Do resuscitation attempts in children who die, cause injury? AB - OBJECTIVE: To determine the incidence, type, and pattern of injury related to resuscitation attempts in children who die. DESIGN: Retrospective review of ambulance, hospital, and necropsy case records. METHOD: All children who died aged 0-14 years between 1994 and 1996, and underwent a full necropsy at the Victorian Institute of Forensic Medicine (Melbourne, Australia) were identified. Children who were subject to recognised trauma before resuscitation or died because of a congenital abnormality were excluded. The records of all remaining children were reviewed. Children were grouped according to whether resuscitation was attempted or not. RESULTS: From a total of 346 children who died, 204 (58.6%) were identified as meeting the inclusion criteria. Resuscitation was performed in 153 (75%) children and was started before ambulance arrival in 123 (60.3%) children. Injuries were detected at necropsy in 65 (42.5%) of children who had resuscitation compared with six (11.7%) of children who had no resuscitation (p<0.0001) chi(2) test. All but two of these injuries were of a minor nature consisting principally of bruises or abrasions. Two significant injuries were identified both occurring as a result of readily identifiable resuscitation procedures. The likelihood of injury increased with the length of resuscitation. In children resuscitated for less than 60 minutes the incidence of injury was 27% compared with 62% for children resuscitated for longer ( p<0.0001). CONCLUSION: This study has shown that cardiopulmonary resuscitation commonly causes minor injuries such as superficial bruises and abrasions and the likelihood of such injury increases with the duration of the cardiopulmonary resuscitation. This information should reassure parents and caregivers that basic life support may be instituted without fear of causing significant injury or adversely affecting outcome in the child with cardiorespiratory arrest. Caution must be exercised when attributing significant injuries to resuscitation attempts and alternative causes must be fully investigated. PMID- 12533360 TI - An evidence and consensus based guideline for the management of a child after a seizure. AB - OBJECTIVE: An evidence and consensus based guideline for the management of the child who presents to hospital having had a seizure. It does not deal with the child who is still seizing. The guideline is intended for use by junior doctors, and was developed for this common problem (5% of all paediatric medical attenders) where variation in practice occurs. OPTIONS: Assessment, investigations (biochemistry, lumbar puncture, serum anticonvulsant levels, EEG in particular), and/or admission are examined. OUTCOMES: The guideline aims to direct junior doctors in recognising those children who are at higher risk of serious intracranial pathology including infection, and conversely to recognise those children at low risk who are safe to go home. EVIDENCE: A systematic review of the literature was performed. Articles were identified using the electronic data bases Medline (from 1966 to June 1998), Embase (from 1980 to June 1998) and Cochrane (to June 1998), and selected if they investigated the specified clinical question. Personal reviews were excluded. Selected articles were appraised, graded, and synthesised qualitatively. Statements of recommendation were made. CONSENSUS: An anonymous, postal Delphi consensus development was used. A national panel of 30 medical and nursing staff regularly caring for these children were asked to grade their agreement with the statements generated. They were sent the relevant original publications, the appraisals, and literature review. On the second and third rounds they were asked whether they wished to re-grade their agreement in the light of other panellists' responses. Consensus was defined as 83% of panellists agreeing with the statement. Recommendations in brief: For afebrile seizures all children should have their blood pressure recorded, but no other investigations are routine although a seizing or somnolent child should have blood glucose measured; all children under 1 year should be admitted. For seizures with fever, clinical signs indicating the need to treat as meningitis are given. Children should be admitted if they are under 18 months old, have had a complex seizure, or after pretreatment with antibiotics. VALIDATION: The guideline has undergone implementation and evaluation in a paediatric accident and emergency department, the results of which will be published separately. Only one alteration was made to the guideline as a result of this validation process, which is included here. PMID- 12533361 TI - Accident and emergency attendances by children under the age of 1 year as a result of injury. AB - OBJECTIVES: To examine all accident and emergency (A&E) department attendances by children under the age of 1 year over a period of 12 months. Also to try to identify the prevalence and severity of accident types in small children and to suggest ways to reduce such accidents. METHODS: The A&E department of the Royal Aberdeen Children's Hospital (RACH) serves a population of over half a million. All children under 1 year of age attending this department in the year 2000 had their case notes reviewed by the author and the cause, type, and severity of the illness or injury noted. RESULTS: During the 12 month audit period 1416 new cases under the age of 1 year presented to RACH, 790 of which presented directly to A&E. Six hundred and eighteen (78%) were self referred and 116 children attended A&E on more than one occasion during the year. Four hundred and thirty four (55%) of the A&E attendances were classed as "accidents", the remainder were mainly for medical conditions such as respiratory distress. Two hundred and sixty four (61%) were caused by falls and 38% were admitted for inpatient management. Two hundred and twenty nine (29%) required radiographs, which revealed 30 fractures. Thirty seven children sustained scalds/burns and there were 33 accidental ingestions. Six cases were judged to be non-accidental. CONCLUSIONS: There is a surprisingly high rate of "accidental" injury in this age group, bringing into question the effectiveness of current accident prevention strategies. Perhaps specific prevention advice should be targeted at parents and carers of young children. There should always be a high index of suspicion for non-accidental injury. PMID- 12533362 TI - Comprehensive drug screening in decision making of patients attending the emergency department for suspected drug overdose. AB - OBJECTIVES: This study aimed to evaluate the usefulness of a comprehensive drug screening method as a first line diagnostic tool on clinical decision making in patients attending an emergency department for suspected drug overdose in terms of agreement between physicians on patients' disposal. METHODS: Five emergency physicians retrospectively evaluated the records of 142 adult patients, admitted to the emergency department of a community hospital for suspected drug overdose. They were asked for an expert opinion on patients' disposal at the end of the observation period, based on paired records, with/without the results of a comprehensive drug screening. RESULTS: In the absence of the drug screening, a very poor agreement (kappa statistics) was observed between physicians. When the drug screening was available, the interobserver agreement for decision on patients' disposal increased to the fair to good range (global agreement: from 0.238 (0.019) to 0.461 (0.020) (mean(SE)); p<0.001). The agreement also increased when admission to an intensive care unit, to a general ward, and discharge from hospital were separately analysed. The availability of drug screening would have saved 21.7% of hospital admissions and 53.3% of high dependency and/or intensive care unit admissions. CONCLUSION: Comprehensive drug screening adds to decision making for patients attending an emergency department for suspected drug overdose, improving agreement among physicians on patients' disposal and potentially saving hospital resources. PMID- 12533363 TI - Emergency department investigation of deep vein thrombosis. AB - OBJECTIVES: To apply a previously validated clinical model for predicting pre test probability of deep vein thrombosis (DVT) to patients attending an emergency department with symptoms suggestive of DVT and assess its reproducibility in the patient population. To measure the diagnostic value of the SimpliRED D-dimer assay in effectively excluding DVT. METHOD: Prospective study between March 1999 and October 2000 of adult patients attending with suspected DVT. Patients were assessed using Wells' clinical prediction tool and risk stratified. SimpliRED D dimer estimations were then performed and compression ultrasonography arranged. The pre-test probabilities of DVT in the low, moderate, and high risk groups of Wells' original cohort were compared with the authors' figures and the sensitivity, specificity, and predictive values of the SimpliRED assay calculated for the patient population. RESULTS: Application of Wells' criteria to patients in the department permitted stratification into high, moderate, and low risk groups (prevalence of DVT 58.3%, 8.9%, and 1.5% respectively). SimpliRED D-dimer assay sensitivity was 63.4% with specificity of 74.8%, with a likelihood ratio of 2.52 for a positive test and 0.49 for a negative test. CONCLUSIONS: Clinical risk stratification allowed patients to be categorised into high, moderate, and low risk groups, albeit with less discriminatory power than originally described by Wells. The low sensitivity of the SimpliRED D-dimer assay when used routinely in a busy emergency department raises substantial doubt over the use of this test to rule out DVT, even in low risk patients. PMID- 12533364 TI - Prospective randomised trial comparing traditional suture technique with the dynamic sliding loop suture technique in the closure of skin lacerations. AB - OBJECTIVE: The aim of this study was to compare the cosmetic appearance and related complications of selected skin lacerations closed by traditional suture technique with that of the dynamic sliding loop suture technique. DESIGN: Prospective, randomised clinical trial. SETTING: A district general hospital. PARTICIPANTS: Thirty seven eligible patients aged between 16 and 60 years with skin lacerations (no deeper than superficial fascia) to the limbs, trunk, and neck (excluding face and scalp) and with no associated neurovascular or tendon injuries were recruited into the trial. The exclusion criteria used included immunocompromised patients (diabetics, malignancy, renal failure, corticosteroid treatment), primary dermatological conditions (psoriasis, eczema), keloid prone or susceptible patients, and wounds judged unsuitable for primary closure. INTERVENTION: Skin lacerations were randomly allocated to have closure by either the traditional method or by use of the dynamic sliding loop suture method. The trial had ethical approval of the hospital ethics committee and all participants were fully informed of the trial both verbally and by an information leaflet. Written informed consent was obtained before starting the study. MAIN OUTCOME MEASURE: The cosmetic appearance of photographs of wounds immediately after suture removal and at three months were assessed by a general surgeon and an orthopaedic surgeon blinded to the technique used. The 10 point visual analogue cosmesis scale was used for scoring the appearance of the wounds. SECONDARY OUTCOME MEASURE: The presence of wound closure related complications such as infection, dehiscence, suture slippage, wound edge submergence, skin edge necrosis, and haematoma formation were noted and recorded on follow up visits. RESULTS: A total of 37 patients participated in the trial over a period of 18 months. Seventeen patients underwent suture by the traditional technique and 20 by the dynamic sliding loop suture technique. Four patients (all from the traditional group) were lost to follow up and not included in the study. Most wounds healed uneventfully although there was one case of wound infection in each group. Four patients had ligature slippage (three from the dynamic suture and one from the traditional group). Two patients had evidence of skin edge necrosis both from the traditional technique group. A comparison of the healed wounds by two independent assessors blinded to the technique used showed no statistically significant difference observed between the two types of intervention (Wilcoxon matched paired test p>0.05) immediately after suture removal or at three months. Although there was no statistically demonstrable difference, the comparative paired absolute visual analogue scores seemed to consistently show higher values for the sliding loop technique. CONCLUSION: The cosmetic appearance of wounds sutured using the dynamic sliding loop suture technique in this study were not statistically shown to be superior to those sutured using the traditional suture technique although absolute visual analogue scores consistently showed higher values for the sliding loop technique. The significance of this is unclear and may warrant a much larger trial to see if it is a trend that a larger population of participants can help to support or refute with regard to the superiority of this new technique. PMID- 12533365 TI - Management of soft tissue knee injuries in an accident and emergency department: the effect of the introduction of a physiotherapy practitioner. AB - OBJECTIVE: To assess the effect of the introduction of a physiotherapist with an extended scope of practice in the management of acute soft tissue knee injuries in an accident and emergency (A&E) department. METHODS: The A&E department established an Acute Knee Screening Service (AKSS) in 1998. An experienced physiotherapist was appointed to run the AKSS after additional training. Local guidelines and protocols were developed in conjunction with trauma knee surgeons, radiologists, physiotherapists, and A&E doctors. This study describes 100 consecutive patients managed by the AKSS over a three month period and comparisons are made over a similar period before the introduction of the AKSS. RESULTS: The initial diagnosis of patients with acute knee injuries referred to the service showed meniscal injuries (38%), cruciate ligament injuries (18%), fractures (2%), patellofemoral joint injuries (10%), and others (32%). Ninety five per cent of patients referred to AKSS were seen within one week. Medical time was saved in both A&E and trauma clinic. Fifty nine per cent of patients were treated and discharged from the service without further medical review. Thirty nine per cent (39) were referred to trauma clinic and of these 44% (17) had MRI scans performed as requested by the physiotherapist. Eighty eight per cent (15) of these scans showed significant abnormality: (nine, anterior cruciate ligament tears, one, posterior cruciate ligament tears, and nine meniscal tears). CONCLUSIONS: A physiotherapy practitioner working with an extended role is a valuable addition to an A&E department. The AKSS improves the quality of care of acute knee injuries, saves medical time, and fosters cooperation across services within the NHS. PMID- 12533366 TI - What is the effect of reporting all emergency department radiographs? AB - OBJECTIVES: To evaluate the effect of formal radiological reporting of all emergency department (ED) radiographs on clinical practice and patient outcome, and to consider whether a selective reporting policy might prove safe and effective. METHODS: All radiographs taken in a single ED over a six month period were prospectively studied simultaneously in both the emergency and radiology departments to detect cases where a radiograph that was considered normal by ED staff was then reported as abnormal by the reporting radiologist. Whenever such a discrepancy occurred the patient's records were scrutinised to ascertain the source of the discrepancy, with a gold standard interpretation derived from senior clinical review and additional investigations where indicated. The clinical impact of the radiologist's formal report was then assessed. Accuracy of interpretation was considered in relation to the grade of ED staff and the radiographic examination obtained. RESULTS: During the study period, 19468 new patient attendances to the ED generated 11749 radiographic examinations. Discrepancies were detected in 175 patients (1.5% of all radiographic examinations). Of these, 136 (1.2%) were subsequently shown to have been incorrectly interpreted in the ED (ED false negatives), with 40 patients (0.3%) undergoing a change in management as a result. In the remaining 39 the ED interpretation was judged to be correct (radiology false positives), with 16 patients undergoing further investigations or visits to the ED to confirm this. CONCLUSIONS: The formal reporting of ED radiographs by the radiology department detects a number of clinically important abnormalities that have been overlooked. However, this formal reporting also generates a number of incorrect interpretations that may lead to further unnecessary investigations. Some groups of ED radiographs (such as those interpreted by an ED consultant and films of the fingers and toes) may not require formal radiological reporting. The adoption of a selective reporting policy may reduce the reporting workload of the radiology department without compromising patient care. PMID- 12533367 TI - Role of the accident and emergency review clinic in UK service provision. AB - OBJECTIVES: Selected patients attending accident and emergency (A&E) are seen again in the A&E review clinic for planned follow up. Despite the large number of patients being seen in these clinics, this area of A&E service provision has rarely been studied. The aim of this study was to determine the current provision of review clinic services in UK A&E departments, their organisation, and their perceived role in current practice. METHODS: Postal survey of all major UK adult and paediatric A&E units. RESULTS: Almost all major UK A&E departments have a review clinic service and almost three quarters see less than 10% as reviews in line with current guidance. Many departments feel their clinics are well run and fulfil their objectives. Others are under pressure from external sources to see more patients. Over a third of departments have no written policies or protocols guiding referral to these clinics. CONCLUSION: Review clinics are an important part of the A&E service in the UK and minimum standards need to be set in the provision of these services. Further audit and research needs to be carried out on the review clinic service to guide its future development. PMID- 12533368 TI - Impact of a newly opened prison on an accident and emergency department. AB - OBJECTIVE: To determine the impact of a newly opened prison on an accident and emergency (A&E) department. METHOD: A new category B prison opened in April 1999, the first privately run prison in Scotland and the third largest in population. All prisoners referred to the A&E department for treatment were identified prospectively during the first year after the opening of the prison. RESULTS: 99 prisoners and four members of staff attended during the one year period. Ages ranged from 18-64 years with a mean age of 29.8 years. Presentations were as a result of deliberate self harm (22%), injury after violence (18%), sports injury (15%), surgical condition (15%), medical illness (13%), accidental injury (9%), ENT problem (2%), and miscellaneous (6%). Thirty seven prisoners (35.6%) were admitted to the hospital. Further review at outpatient clinics was arranged for 15 prisoners. One prisoner died, the result of suicide by hanging. The remaining prisoners were returned to the prison for further management by the prison medical and nursing team. Twelve prisoners re-attended a total of 37 times, ranging from twice to a maximum of eight visits. Some 42.3% of attendances were during "working hours" (09.00-17.00) and 57.7% attended "out of hours" (17.00 09.00). Twenty four referrals (23.1%) were deemed inappropriate by the prison medical team on retrospective review. Sixteen of these occurred "out of hours". Forty one prisoners (39.4%) were known to have a history of injecting drug misuse. Including re-attenders, 59 presentations (56.7%) to the A&E department had a history of injecting drug misuse. Of these 41 prisoners, 11 (26.8%) were hepatitis C positive, with eight of these having a positive polymerase chain reaction test. No prisoners had HIV and only one prisoner was hepatitis B positive. CONCLUSION: The opening of the prison resulted in only a slight increase in the workload of the A&E department. A significant proportion of prisoners were admitted to the hospital highlighting the practical and logistical problems of managing people restrained and in custody. Most cases can be safely referred back to the prison. Increased input is required from the prison medical team when dealing with deliberate self harm, frequent attenders, and "out of hours" referrals. All A&E staff must be aware of the increased risk of hepatitis C infection when dealing with a confined prison population. PMID- 12533369 TI - Patient knowledge of thrombolysis in acute myocardial infarction. AB - A convenience sample of 50 patients admitted for treatment or ruling out of acute myocardial infarction were asked about their knowledge of thrombolysis. Some 44 of 50 knew nothing of thrombolysis and interestingly 30 of these 44 had been previously diagnosed as having ischaemic heart disease. Greater knowledge of the benefit and timeliness of thrombolysis may provide an important incentive for earlier presentation of this group. PMID- 12533370 TI - Observational research methods. Research design II: cohort, cross sectional, and case-control studies. AB - Cohort, cross sectional, and case-control studies are collectively referred to as observational studies. Often these studies are the only practicable method of studying various problems, for example, studies of aetiology, instances where a randomised controlled trial might be unethical, or if the condition to be studied is rare. Cohort studies are used to study incidence, causes, and prognosis. Because they measure events in chronological order they can be used to distinguish between cause and effect. Cross sectional studies are used to determine prevalence. They are relatively quick and easy but do not permit distinction between cause and effect. Case controlled studies compare groups retrospectively. They seek to identify possible predictors of outcome and are useful for studying rare diseases or outcomes. They are often used to generate hypotheses that can then be studied via prospective cohort or other studies. PMID- 12533371 TI - Reduction of pulled elbows. PMID- 12533373 TI - Alternative treatments for neck sprain. AB - A short cut review was carried out to establish whether osteopathy or chiropractic treatments improve outcome in patients with neck sprain. Altogether 206 papers were found using the reported search, of which nine presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. A clinical bottom line is stated PMID- 12533374 TI - Tetanus prophylaxis in superficial corneal abrasions. AB - A short cut review was carried out to establish whether tetanus prophylaxis is indicated after non-penetrating corneal abrasion. Altogether 30 papers were found using the reported search, of which one presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of this best paper are tabulated. A clinical bottom line is stated. PMID- 12533375 TI - Gastric lavage in tricyclic antidepressant overdose. AB - A short cut review was carried out to establish whether gastric lavage is indicated after tricyclic antidepressant overdose. Altogether 82 papers were found using the reported search, of which one presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of this best paper are tabulated. A clinical bottom line is stated. PMID- 12533376 TI - Treatment of uncomplicated subungual haematoma. AB - A short cut review was carried out to establish whether nail removal and nail bed repair is better than simple trephining in patients with significant subungual haematoma. Altogether 312 papers were found using the reported search, of which four presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 12533377 TI - Use of sterile gloves in the treatment of simple wounds. AB - A short cut review was carried out to establish whether the use of sterile gloves during the treatment of simple wounds reduces infections. Altogether 48 papers were found using the reported search, but none presented any evidence to answer the clinical question. More research is needed in this area and, in the mean time, local advice should be followed. PMID- 12533378 TI - Contraindications to thrombolysis in patients taking coumarins. AB - A short cut review was carried out to establish what level of INR was a contraindication to thrombolysis in patients taking warfarins. Altogether 296 papers were found using the reported search, but none presented any evidence to answer the clinical question. More research is needed in this area and, in the mean time, local advice should be followed. PMID- 12533379 TI - Antibiotics in orbital floor fractures. AB - A short cut review was carried out to establish whether prophylactic antibiotics are indicated in patients with undisplaced maxillary or orbital floor fractures. Altogether 214 papers were found using the reported search, but none presented any evidence to answer the clinical question. More research is needed in this area and, in the mean time, local advice should be followed. PMID- 12533380 TI - Article 12. Major incidents, leadership, and series summary and review. AB - This last article in the series focuses on the impact of a major incident in the emergency department, leadership qualities, and summarises the topics discussed in the previous papers. PMID- 12533381 TI - Interventions for post-traumatic stress disorder and psychological distress in emergency ambulance personnel: a review of the literature. AB - A literature review was carried out to establish the extent of the literature on interventions for psychological distress and post-traumatic stress disorder in emergency ambulance personnel. A total of 292 articles were identified. Of these, 10 were relevant to this review. The primary intervention used with this population was critical incident stress debriefing, although there was some debate in the literature about the effectiveness of this intervention and the quality of the research conducted. More high quality research is needed on critical incident stress debriefing before being confident of its effectiveness. PMID- 12533382 TI - Post-traumatic stress among Swedish ambulance personnel. AB - OBJECTIVE: Emergency workers, including ambulance personnel, must cope with a variety of duty related stressors including traumatic incident exposures. Little is known about the variables that might be associated with post-traumatic stress symptom in high risk occupational groups such as ambulance personnel. This study investigated the prevalence of post-traumatic stress disorder among Swedish ambulance personnel. METHODS: To estimate the prevalence of trauma related disorders, a representative group of 362 ambulance personal from the county of Vastra Gotaland in Sweden was surveyed through use of a Swedish version of Antonovsky's 13-item short version of Sense of Coherence Scale, to measure reactions to traumatic events two instruments were used, Impact of Event Scale (IES-15) and the Post Traumatic Symptom Scale (PTSS-10). A total of 223 of the ambulance personnel reported that they had had experience of what they described as traumatic situations. RESULTS: Of those who reported a traumatic situation 15.2% scored 31 or more on the IES-15 sub scale. Scores over 31 indicate a stress reaction with certain likelihood of post-traumatic disorder. On the PTSS-10 subscale 12.1% scored 5 or more, which indicates a relative strong reaction. The study indicates that lower sense of coherence predicts post-traumatic stress. Other predictors for the extent of traumatic stress were longer job experience, age, physical and psychological workload. CONCLUSIONS: The high prevalence of post-traumatic stress disorder symptoms in ambulance personnel indicates an inability to cope with stress in daily work. The strong relation between post traumatic stress and Sense of Coherence Scale may be useful in predicting vulnerability for post-traumatic symptoms among recently employed ambulance service personnel. To prevent or reduce the upcoming of post-traumatic stress disorder symptoms it must be possible to take leave of absence, or for a longer or shorter time be transferred to non-emergency duties. This study presents a better understanding between post-traumatic stress and underlying factors among ambulance personnel. PMID- 12533383 TI - Is the public equipped to act in out of hospital cardiac emergencies? AB - STUDY OBJECTIVE: This study aimed to determine whether the people in Australia are informed about and prepared to intervene in a cardiac emergency. METHODS: A cross sectional telephone survey, which contained sections regarding participant demographics, cardiopulmonary resuscitation (CPR) training, knowledge of CPR, and the emergency contact number and potential barriers to performing chest compressions and mouth to mouth. RESULTS: A total of 1489 people completed the questionnaire. Only 11% of the population had recently (<12 months) trained in CPR. When presented with a cardiac arrest scenario most participants stated that they would telephone 000. Significantly more respondents believed that they would give mouth to mouth to a family member compared with a stranger. A bleeding victim and fear of not having the skills were the most common barriers that reduced the participants perceived willingness to perform chest compressions and mouth to mouth. CONCLUSION: This study suggests that a low percentage of the public is currently trained in CPR and also that they are unprepared to act in a cardiac emergency. PMID- 12533384 TI - Prehospital use of paracetamol among children attending the accident and emergency department. AB - OBJECTIVES: To identify patterns of prehospital administration of paracetamol to children who were perceived to be feverish by their carers. METHODS: A prospective cohort study of carers of children attending a paediatric accident and emergency (A&E) department. Carers of children completed a questionnaire to identify domestic patterns of paracetamol use. Data were collected on temperature of the child in the A&E department, administration of antipyretics in the A&E department, diagnosis, and disposal from the A&E department. RESULTS: Seventy five adults attending the A&E department consented to involvement. Sixty five of the children were feverish on arrival in the A&E department. Twenty one children (32.3%) had not received paracetamol before attending. There was a significant relation between knowledge of the antipyretic properties of paracetamol and administration (chi(2)=5.0, p<0.05). There was a significant correlation between fever and administration of paracetamol in the A&E department (chi(2)=23.7, p<0.01), however, 15 feverish patients (24.6%) were not treated. CONCLUSIONS: Most carers administer paracetamol appropriately in the prehospital setting. Administration correlates significantly with knowledge of its benefits. There is scope for education of carers and A&E department staff in the in the appropriate use of antipyretics such as paracetamol. PMID- 12533385 TI - Superior dislocation of the patella; a case report and review of the literature. AB - Superior dislocation of the patella is a rare diagnosis. A 72 year old woman attended the accident and emergency department of the hospital with a painful right knee after a knock to the knee. Clinical examination and radiographs confirmed a superior dislocation of the patella, which was reduced and closed with the aid of simple analgesia. The authors present the case report and discuss the relevant literature. PMID- 12533387 TI - Delayed diagnosis of foreign body aspiration in children. AB - Foreign body aspiration in children is common and usually presents with an initial episode of choking with subsequent respiratory symptoms. There may be cough, wheeze, or stridor, with decreased or abnormal breath sounds on examination. However, it can mimic other illnesses and cause difficulty in diagnosis. Radiological investigations may help to confirm aspiration but should not be used to exclude it. Three cases are presented of foreign body aspiration with a delay in diagnosis ranging from days to weeks. It is believed that delay could have been avoided with a more careful approach to the history and more appropriate use of investigations. These cases demonstrate that children with a history of choking and subsequent symptoms should be referred for bronchoscopy. PMID- 12533386 TI - Haemodynamic and electrocardiographic consequences of severe nicorandil toxicity. AB - A 35 year old woman was admitted to the emergency department two hours after ingesting 60x20 mg tablets of nicorandil, total 1.2 g. The dominant feature of icorandil toxicity was profound peripheral vasodilatation associated with coronary hypoperfusion. Despite widespread electrocardiographic signs of myocardial ischaemia, there was no evidence of myocardial damage and no serious cardiac arrhythmia. Volume loading and pressor support proved to be an effective treatment strategy. PMID- 12533388 TI - Haemoptysis as a late presentation of aneurysm leakage after aortic coarctation repair: the case for vigilant lifelong radiology surveillance? PMID- 12533389 TI - First branchial cleft anomaly presenting as a recurrent post-auricular abscess. AB - Embryological anomalies of the first branchial cleft are uncommonly encountered. They usually present as cysts, swellings, or fistulas in the pre-auricular or post-auricular area or high in the neck, which may become infected. Failure to recognise these unusual cases may result in misdiagnosis, inadequate treatment, and subsequent recurrence. Further definitive surgery may thus be complicated. A case is reported of a patient who attended accident and emergency on three occasions with an infected post-auricular cyst, which was treated by incision and drainage. It was subsequently found to be a first branchial cleft anomaly. PMID- 12533390 TI - Scrotal pain in the absence of torsion; need for vigilance. AB - Epididymitis is a common presentation of acute testicular pain seen in the emergency department, the differential diagnosis being testicular torsion. The vast majority of young men with epididymitis have an infective aetiology and this settle with antibiotic treatment. The clinical course of a patient who presented with testicular pain is described. At ultrasonography, the patient was found to have the uncommon condition of testicular microlithiasis, a condition that has been linked to malignant disease. Emergency doctors should be aware of the potential consequences of returning scrotal pain consistent with epididymitis to the community on antibiotic treatment alone. All patients with probable epididymitis should have either a scrotal ultrasound or specialist follow up. PMID- 12533391 TI - Unusual presentation of atrial fibrillation. AB - A case is reported of atrial fibrillation in a young healthy man after head injury and the possible causes are discussed. The atrial fibrillation reverted spontaneously to normal rhythm in two days. The authors are not aware of a similar report in the literature. PMID- 12533392 TI - Use of cut endotracheal tubes should be avoided in the initial resuscitation of the burned patient. PMID- 12533393 TI - Comminuted fracture of the talus not visible on the initial radiograph. AB - Fractures of the talus are rare injuries and fractures of the body of the talus are particularly rare. Diagnosis of these fractures is also difficult as initial radiographs may be normal, particularly with osteochondral talar dome fractures. Long term morbidity is common after fractures of the talus. A case is presented of a patient with a comminuted fracture of the body of the talus with non diagnostic initial standard ankle radiographs. Accident and emergency doctors should be aware of this injury, and be suspicious that patients with an appropriate mechanism of injury and pronounced pain may require further investigation despite normal standard ankle radiographs, as an occult fracture of the talus may be present. PMID- 12533394 TI - Fracture of lateral process of the talus presenting as ankle pain. AB - The case is presented of a 27 year old woman with lateral ankle pain after an inversion injury sustained while dancing. Although initial radiographs failed to identify the fracture, radiographs of the ankle at six weeks showed an unsuspected fracture of the lateral process of the talus. The fracture was treated with cast immobilisation for six weeks and the patient is currently undergoing aggressive physiotherapy. A literature review revealed that fractures of the lateral process of the talus are frequently overlooked and should be considered in the differential diagnosis of patients with acute and chronic ankle pain as an early diagnosis and treatment prevent long term complications. PMID- 12533395 TI - The yolkless egg and the evolution of eutherian viviparity. AB - All vertebrate follicles have the same basic structure. Viviparity also occurs in all vertebrates except birds, but it is the only form of reproduction in eutherians ("placental mammals"). Their mature follicles are vesicular, and their oocytes are yolkless. Clues to the origin of these unique characteristics are in the incidence of atresia and the role of yolk in reproduction. In broadcast spawning, atresia is as rare as it is common among eutherians and other nonspawning vertebrates. In all but the eutherians, at least the initial-and in most cases all-stages of embryogenesis depend crucially on the zygote's yolk. Eutherian reproduction, therefore, must have evolved in connection with genetic changes that caused fragility of the oocyte, instability of the follicle, and loss of the ability to produce vitellogenin (VTG), the main lipoprotein of yolk. Mutations can result in adaptations by uncovering hidden properties in a trait and/or its environment. Useful mutations in recessive alleles can spread through a population as heterozygotes, invisible until the number of homozygotes for the mutation is large enough for them to suddenly appear and form the nucleus of a new breeding population. Such a mutation probably truncated a long, oviductal based, aplacental gestation of a small, lightly yolked zygote in an endothermic, mammal-like reptile and converted it into an early monotreme or marsupial-like mammal (pantothere). Against tremendous odds, another mutation later caused loss of the genes for VTG. The resultant yolkless zygote survived because 1). the mutation also affected a network of homeiotic genes controlling the ontogeny of the entire reproductive system and 2). the system contained enough hidden properties for the mutation to change the character of the oocyte, its granulosa cells and corpus luteum, the zygote, and the uterus in a way that virtually assured the new zygote's survival. Eutherian reproduction, however, is neither better nor worse than other forms; it is only different. PMID- 12533396 TI - Evolution and nomenclature of the zona pellucida gene family. AB - Three subfamilies of genes are acknowledged within the zona pellucida (ZP) gene family. At present, these subfamilies each have two names that are used interchangeably: ZPA or ZP2, ZPB or ZP1, and ZPC or ZP3. The ZPA genes encode the longest protein sequences and the ZPC genes the shortest. Recently, several sequences, which have no clear relationship to the three subfamilies, have been identified. These sequences include two paralogous ZP genes from Xenopus laevis and a single gene from the fish Oryzias latipes. We have conducted extensive phylogenetic analyses of the known ZP genes. As well as establishing the evolutionary relationships among these genes, the analyses make it clear that the dual nomenclature system is no longer feasible, because major paralogous groups are present in the ZPB (ZP1) family of genes of amniotes. We propose a unified system of nomenclature for the ZP gene family that removes the existing ambiguities. PMID- 12533397 TI - Testosterone-dependent effects of galanin on pituitary luteinizing hormone secretion in male rats. AB - Galanin is a 29-amino-acid peptide that colocalizes with GnRH in hypothalamic neurons. High concentrations of galanin are present in portal vessel blood of both male and female rats, and galanin receptors are present on gonadotropes in both sexes. Results from studies of female rats indicate that galanin acts at the level of the pituitary to directly stimulate LH secretion and also to enhance GnRH-stimulated LH secretion. The effects of galanin on pituitary LH secretion in male rats are relatively uncharacterized; thus, the present in vivo study was conducted 1). to examine the ability of galanin to affect basal or GnRH stimulated LH secretion in male rats and 2). to determine whether the effects of galanin on LH secretion in male rats are testosterone-dependent. All three doses of galanin used (1, 5, and 10 micro g/pulse) significantly enhanced GnRH stimulated LH secretion in intact male rats. Only the highest dose of galanin directly stimulated LH secretion (without GnRH coadministration) in intact males. Galanin did not directly stimulate LH secretion or enhance GnRH-stimulated LH secretion in castrated male rats. In fact, the highest dose of galanin inhibited GnRH-stimulated LH secretion in castrated males. Upon testosterone replacement, the ability of galanin to directly stimulate LH secretion and to enhance GnRH stimulated LH secretion was restored in castrated males. These results suggest a role for galanin in the regulation of LH release in male rats and demonstrate that testosterone upregulates the ability of the pituitary to respond to the stimulatory effects of galanin. PMID- 12533398 TI - Estrogen synthesis in fetal sheep brain: effect of maternal treatment with an aromatase inhibitor. AB - The aim of the present study was to determine whether the fetal lamb brain has the capacity to aromatize androgens to estrogens during the critical period for sexual differentiation. We also determined whether administration of the aromatase-inhibitor 1,4,6-androstatriene-3,17-dione (ATD) could cross the placenta and inhibit aromatase activity (AA) in fetal brain. Eight pregnant ewes were utilized. On Day 50 of pregnancy, four ewes were given ATD-filled Silastic implants, and the other four ewes received sham surgeries. The fetuses were surgically delivered 2 wk later (Day 64 of gestation). High levels of AA (0.8-1.4 pmol/h/mg protein) were present in the hypothalamus and amygdala. Lower levels (0.02-0.1 pmol/h/mg protein) were measured in brain stem regions, cortex, and olfactory bulbs. The Michaelis-Menten dissociation constant (K(m)) for aromatase in the fetal sheep brain was 3-4 nM. No significant sex differences in AA were observed in brain. Treatment with ATD produced significant inhibition of AA in most brain areas but did not significantly alter serum profiles of the major sex steroids in maternal and fetal serum. Concentrations of testosterone in serum from the umbilical artery and vein were significantly greater in male than in female fetuses. No other sex differences in serum steroids were observed. These data demonstrate that high levels of AA are found in the fetal sheep hypothalamus and amygdala during the critical period for sexual differentiation. They also demonstrate that AA can be inhibited in the fetal lamb brain by treating the mother with ATD, without harming fetal development. PMID- 12533399 TI - Embryotrophic factor-3 from human oviductal cells affects the messenger RNA expression of mouse blastocyst. AB - Our previous results showed that embryotrophic factor-3 (ETF-3) from human oviductal cells increased the size and hatching rate of mouse blastocysts in vitro. The present study investigated the production of ETF-3 by an immortalized human oviductal cell line (OE-E6/E7) and the effects of ETF-3 on the mRNA expression of mouse embryos. The ETF-3 was purified from primary oviductal cell conditioned media using sequential liquid chromatographic systems, and antiserum against ETF-3 was raised. The ETF-3-supplemented Chatot-Ziomek-Bavister medium was used to culture Day 1 MF1 x BALB/c mouse embryos for 4 days. The ETF-3 treatment significantly enhanced the mouse embryo blastulation and hatching rate. The antiserum, at concentrations of 0.03-3%, abolished the embryotrophic effect of ETF-3. Positive ETF-3 immunoreactivity was detected in the primary oviductal cells, OE-E6/E7, and blastocysts derived from ETF-3 treatment. Vero cells (African Green Monkey kidney cell line), fibroblasts, and embryos cultured in control medium did not possess ETF-3 immunoreactivity. The mRNA expression patterns of the treated embryos were studied at the blastocyst stage by mRNA differential display reverse transcription-polymerase chain reaction (DDRT-PCR). The DDRT-PCR showed that some of the mRNAs were differentially expressed after ETF-3 treatment. Twelve of the differentially expressed mRNAs that had high homology with cDNA sequences in the GenBank were selected for further characterization. The differential expression of seven of these mRNAs (ezrin, heat shock 70-kDa protein, cytochrome c oxidase subunit VIIa-L precursor, proteinase-activated receptor 2, eukaryotic translation initiation factor 2beta, cullin 1, and proliferating cell nuclear antigen) was confirmed by semiquantitative RT-PCR. In conclusion, immortalized oviductal cells produce ETF 3, which influences mRNA expression of mouse blastocyst. PMID- 12533400 TI - Quantification of histone acetyltransferase and histone deacetylase transcripts during early bovine embryo development. AB - Mammalian oocytes are very unique cells with an unlimited developmental potential. These totipotent cells are able to remove existing gene-expression patterns and to impose new ones. However, genome reprogramming is still a mystery. Posttranslational modifications by acetylation of the N-termini portion of histones composing the nucleosome are involved in genome reprogramming. These modifications alter the higher-order chromatin structure to render the DNA accessible to the regulatory and transcriptional machinery. In the present study, we have investigated, to our knowledge for the first time, precise expression patterns of seven genes involved in chromatin structure throughout bovine embryo development. Oocytes harvested from bovine ovaries were used for in vitro production of germinal vesicle oocytes, metaphase II oocytes, 2- and 8-cell embryos, and blastocysts. Total RNA was extracted from pools (triplicates) of 20 oocytes or from embryos of each developmental stage. By means of quantitative reverse transcription-polymerase chain reaction using SYBR Green to detect double stranded DNA, mRNA expression profiles for histone deacetylases (HDAC1, HDAC2, HDAC3, and HDAC7), histone acetyltransferases (GCN5 and HAT1), and histone H2A were established. Transcripts for all genes were detected at all stages from the oocyte to the blastocyst. The HDAC1, HDAC2 (class I HDAC), and HAT1 (type B HAT) revealed similar expression profiles. The HDAC3 (class I HDAC) tends to have an expression profile similar to those of HDAC1, HDAC2, and HAT1, whereas the HDAC7 (class II HDAC) and GCN5 (type A HAT) profiles were different from those three. These results indicate variable levels of histone deacetylases and histone acetyltransferases throughout embryonic development and may indicate the ones that are involved in somatic remodeling. PMID- 12533401 TI - Effects of perinatal maternal food restriction on pituitary-gonadal axis and plasma leptin level in rat pup at birth and weaning and on timing of puberty. AB - The effects of maternal 50% food restriction (FR) during the last week of gestation and/or lactation on pituitary-gonadal axis (at birth and weaning), on circulating levels of leptin (at weaning), and on the onset of puberty have been determined in rats at birth and at weaning. Maternal FR during pregnancy has no effect at term on the litter size, on the basal level of testosterone in male pups, and on the drastic surge of circulating testosterone that occurs 2 h after birth. At weaning, similar retardation of body growth is observed in male and female pups from mothers exposed to FR. This undernutrition induces the most drastic effects when it is performed during both gestation and lactation or during lactation alone. Drastic retardation of testicle growth with reduction of cross-sectional area and intratubular lumen of the seminiferous tubules is observed in male pups from mothers exposed to undernutrition during both gestation and lactation or during lactation alone. Maternal FR during the perinatal period reduces circulating levels of FSH in male pups without affecting LH and testosterone concentrations. Maternal FR does not affect circulating levels of LH, estradiol, and progesterone in female pups. Female pups from mothers exposed to FR during both gestation and lactation show a significant increase of plasma FSH as well as a drastic retardation of ovarian growth. The follicular population was also altered. The number of antral follicles of small size (vesicular follicles) was increased, although the number of antral follicles of large size (graafian follicles) was reduced. Maternal FR occurring during both late gestation and lactation (male and female pups), during lactation alone (male and female pups), or during late gestation (female pups) induces a drastic reduction of plasma leptin and fat mass in pups at weaning. The onset of puberty is delayed in pups of both sexes from mothers exposed to FR during lactation and during both gestation and lactation. In conclusion, these data demonstrate that a perinatal growth retardation induced by maternal FR has long-term consequences on both size and histology of the genitals, on plasma gonadotropins and leptin levels, on fat stores at weaning, and on the onset of puberty. PMID- 12533402 TI - Loss of cyclooxygenase-2 retards decidual growth but does not inhibit embryo implantation or development to term. AB - Previous reports have described that female mice deficient in cyclooxygenase-2 (COX2) are largely infertile because of failure to ovulate, poor fertilization, and defective implantation and decidualization. In the present study, we reinvestigated reproduction in these mice and found they do show a reduction in the numbers of ovulated and fertilized eggs. However, we did not observe any substantial effect on embryo implantation frequencies or an inability of COX2 deficient females to support embryo development to weaning. Pseudopregnant COX2 null recipients do not show any alteration in the timing of implantation following blastocyst transfer, but they do show a delay in the initial rate of decidual growth after implantation that lags by approximately 24 h compared to that in heterozygous or wild-type recipients. These results support previous findings that COX2 has a role in mediating the initial uterine decidual response but is not essential to sustaining decidual growth and embryo development throughout the remainder of pregnancy. PMID- 12533403 TI - Differential splicing of the phospholipid hydroperoxide glutathione peroxidase gene in diploid and haploid male germ cells in the rat. AB - Phospholipid hydroperoxide glutathione peroxidase (PHGPx, 20 kDa) and sperm nuclei glutathione peroxidase (snGPx, 34 kDa) are two selenoproteins present in mammalian testis and epididymal spermatozoa. They originate from the differential splicing of the PHGPx gene and appear to play important roles in sperm physiology. To determine the stages of spermatogenesis in which they are present, we compared the expression pattern of these two enzymes in highly purified populations of germ cells during specific phases of differentiation. In Northern and Western blotting experiments, both PHGPx transcript and protein were markedly expressed in pachytene spermatocytes and round spermatids. In contrast, the testis-specific snGPx was detected at both the mRNA and protein level only in haploid round spermatids. Accordingly, the developmental analysis of testicular RNAs from rats of different ages first revealed the appearance of PHGPx and snGPx transcripts at Day 20 and Day 30, respectively. Furthermore, both meiotic and postmeiotic cells contained catalytically active PHGPx/snGPx, with higher activity in the haploid cells. The intracellular distribution of PHGPx in mitochondria and nuclei of meiotic cells was demonstrated by immunocytochemical electron microscopy and Western blotting. These findings provide evidence that the PHGPx gene is differentially spliced during the meiotic prophase and haploid cell phases of spermatogenesis. PMID- 12533404 TI - GATA-4 and GATA-6 transcription factors: expression, immunohistochemical localization, and possible function in the porcine ovary. AB - The expression and localization of GATA-4 and GATA-6 mRNAs and proteins were assessed in porcine ovaries at different stages of the estrous cycle. Reverse transcription polymerase chain reaction and Western blot analyses revealed that GATA-4 and GATA-6 transcripts and proteins were strongly expressed in granulosa cells isolated from antral follicles, intact antral follicles, corpora hemorrhagica (CH), and midluteal phase corpora lutea (CL). Immunoblot analyses showed two predominant proteins with molecular masses of approximately 53 and 55 kDa for GATA-4 and one 55-kDa protein for GATA-6. Immunohistochemical studies revealed GATA-4 and GATA-6 nuclear staining in granulosa cells of healthy primordial and primary antral follicles and antral follicle of various sizes. The percentage of immunopositive thecal cell nuclei increased with follicular development. In CH and CL, luteal cells displayed nuclear immunoreactivity for both transcription factors. Regressing CL showed a decrease in GATA immunopositive cells. Immunoreactivity for GATA-4 and GATA-6 was present in most blood vessels. In electrophoretic mobility shift assays, nuclear protein extracts isolated from granulosa cells and CL exhibited both GATA-4 and GATA-6 binding to a GATA consensus oligonucleotide, with GATA-4 the predominant binding protein. GATA-4 and GATA-6 DNA binding was elevated in granulosa cell nuclear extracts from preovulatory (8-10 mm) follicles. Cotransfection of primary cultures of luteinizing granulosa cells with GATA-4 or GATA-6 expression vectors increased the activity of the porcine steroidogenic acute regulatory protein gene promoter significantly but did not significantly activate the inhibin alpha gene promoter. The detection of GATA-4 and GATA-6 mRNAs and proteins in porcine ovaries and the identification of at least one possible target gene may help to establish roles for these GATA factors in follicular development and luteal function. PMID- 12533405 TI - Sonic hedgehog cascade is required for penile postnatal morphogenesis, differentiation, and adult homeostasis. AB - The penis is unique in that it undergoes morphogenesis and differentiation primarily in the postnatal period. For complex structures such as the penis to be made from undifferentiated precursor cells, proliferation, differentiation, and patterning are required. This process involves coordinated activity of multiple signals. Sonic hedgehog (Shh) forms part of a regulatory cascade that is essential for growth and morphogenesis of many tissues. It is hypothesized that the penis utilizes regulatory mechanisms similar to those of the limb and accessory sex organs to pattern penile postnatal morphogenesis and differentiation and that the Shh cascade is critical to this process. To test this hypothesis, Shh, BMP-4, Ptc, and Hoxa-10 localization and function were examined in Sprague-Dawley rat penes by means of quantitative reverse transcription polymerase chain reaction, in situ hybridization, immunohistochemistry, and Western blotting. These genes were expressed in the penis during postnatal morphogenesis in a spatially and temporally restricted manner in adjacent layers of the corpora cavernosal sinusoids. The function of Shh and BMP-4 is to establish and maintain corpora cavernosal sinusoids. The data suggest that Ptc and Hoxa-10 are also important in penile morphogenesis. The continuing function of Shh and targets of its signaling in maintaining penile homeostasis in the adult is significant because disruption of Shh signaling affects erectile function. This is the first report that demonstrates the significant role that Shh plays in establishing and maintaining penile homeostasis and how this relates to erectile function. These studies provide valuable insight that may be applied to improve treatment options for erectile dysfunction. PMID- 12533406 TI - Specific and transient up-regulation of proprotein convertase 6 at the site of embryo implantation and identification of a unique transcript in mouse uterus during early pregnancy. AB - The present investigation was conducted to identify and characterize an mRNA that was found by RNA differential display to be uniquely regulated at the sites of embryo implantation in mouse uterus. This mRNA was upregulated at the sites of blastocyst attachment at implantation and was identified as proprotein convertase 6 (PC6). PC6 mRNA level was low in the nonpregnant and early pregnant uterus before embryo implantation commenced (before Day 4.5, vaginal plug = Day 0). During the initiation and progression of blastocyst attachment (around Day 4.5), the mRNA was dramatically upregulated only at the implantation sites. The increased transcription was maintained on Day 5.5; the mRNA level declined slightly on Day 6.5 and then fell sharply to reach the nonpregnant level around Days 8.5-10.5. Thus, the upregulation is transient and coincides with the period of embryo attachment and implantation; it is also very specific to implantation sites. In situ hybridization analysis localized the mRNA expression predominantly in the decidual cells immediately surrounding the implanting embryo at the antimesometrial pole. Additionally, multiple mRNA species resulting from alternative splicing were observed in the uterus, as previously reported in the intestine and brain, and further analysis of these transcripts identified a uterine-specific PC6 mRNA. These data lead us to suggest that PC6 plays an important role in the processes of stromal cell decidualization and embryo implantation. PMID- 12533407 TI - Rescue of the corpus luteum in human pregnancy. AB - Rescue of the corpus luteum from its programmed senescence maintains progesterone production required for pregnancy. In primates, chorionic gonadotropin produced by the developing conceptus acts as the primary luteotrophic signal. The purpose of this research was to assess corpus luteum rescue by examining changes in daily urinary progesterone metabolite levels during the first week after implantation. We determined the variability in progesterone metabolite profiles and evaluated its relationship to early pregnancy loss in 120 naturally conceived human pregnancies, including 43 early pregnancy losses. In other primates, an abrupt increase in the progesterone metabolite occurs at the time of implantation. This pattern occurred in an estimated 45% of the pregnancies in the present study. In the remaining pregnancies, there was a delayed rise (18%), neither a rise or decline (22%), or a decline (15%) during the week after implantation. The estimated rate of early pregnancy loss increased across these categories (from 5% loss with an abrupt rise at implantation to 100% loss with progesterone metabolite decline). Low urinary hCG levels in early pregnancy were significant determinants of a decline in postimplantation progesterone metabolite. However, preimplantation steroid metabolite levels were not significant, suggesting no inherent problem with the corpus luteum. Examination of individual progesterone metabolite profiles in relation to hCG profiles also indicated that few losses were caused by corpus luteum failure. Delineating the functional importance of an abrupt progesterone rise at the time of implantation may provide new strategies for promoting successful implantation in assisted reproduction. PMID- 12533408 TI - Signal transduction pathways activated by chorionic gonadotropin in the primate endometrial epithelial cells. AB - Successful implantation requires synergism between the developing embryo and the receptive endometrium. In the baboon, infusion of chorionic gonadotropin (CG) modulates both morphology and physiology of the epithelial and stromal cells of the receptive endometrium. This study explored the signal transduction pathways activated by CG in endometrial epithelial cells from baboon (BE) and human (HES). Incubations of BE and HES cells with CG did not significantly alter adenylyl cyclase activity or increase intracellular cAMP when compared with Chinese hamster ovarian cells stably transfected with the full-length human CG/luteinizing hormone (LH) receptor (CHO-LH cells). However, in BE and HES cells, CG induced the phosphorylation of several proteins, among them, extracellular signal-regulated protein kinases 1 and 2 (ERK 1/2). Phosphorylation of ERK 1/2 in uterine epithelial cells was protein kinase A (PKA) independent. This novel signaling pathway is functional because, in response to CG stimulation, prostaglandin E(2) (PGE(2)) was released into the media and increased significantly 2 h following CG stimulation. CG-stimulated PGE(2) synthesis in epithelial cells was inhibited by a specific mitogen-activated protein kinase (MEK 1/2) inhibitor, PD 98059. In conclusion, immediate signal transduction pathways induced by CG in endometrial epithelial cells are cAMP independent and stimulate phosphorylation of ERK 1/2 via a MEK 1/2 pathway, leading to an increase in PGE(2) release as the possible result of cyclooxygenase 2 activation. PMID- 12533409 TI - Chronic administration of anabolic steroids disrupts pubertal onset and estrous cyclicity in rats. AB - Use of anabolic-androgenic steroids (AASs) is becoming increasingly popular among adolescent girls, yet the effects of AASs on female physiology and development are not well understood. The present study compared the effects of chronic exposure to three individual AASs, stanozolol (0.05-5 mg/kg), 17alpha methyltestosterone (0.5-5 mg/kg), and methandrostenolone (0.5-5 mg/kg) on the onset of puberty and estrous cyclicity in the rat. Female rats received daily injections of AASs for 30 days (Postnatal Day [PN] 21-51). Rats receiving the highest dose of each of the AASs (5 mg/kg) displayed vaginal opening at a younger age than rats receiving the oil vehicle. The day of first vaginal estrus was delayed in rats receiving stanozolol (5 mg/kg) or 17alpha-methyltestosterone (0.5 5 mg/kg) but not in rats receiving methandrostenolone. At the highest dose (5 mg/kg), each of the AASs reduced the incidence of regular estrous cyclicity during the treatment period. Concurrent administration (on PN21-51) of the androgen receptor antagonist, flutamide (10 mg/kg, twice daily), reversed the effects of 17alpha-methyltestosterone (5 mg/kg) on vaginal opening. Flutamide administration also eliminated the effects of stanozolol (5 mg/kg) and 17alpha methyltestosterone (5 mg/kg) on the day of first vaginal estrus. In contrast, rats receiving flutamide and methandrostenolone (5 mg/kg) exhibited first vaginal estrus earlier than controls. The present results indicate that chronic exposure to AASs during development has deleterious effects on the female neuroendocrine axis and that these effects appear be mediated via multiple mechanisms. PMID- 12533410 TI - Effect of leptin on progesterone, human chorionic gonadotropin, and interleukin-6 secretion by human term trophoblast cells in culture. AB - Leptin, the 16-kDa protein product of the obese gene, was originally seen as an adipocyte-derived signaling molecule. Recently, it has been suggested to be involved in some functions during pregnancy, particularly in the placenta. In the present study, we investigated the role of leptin in the secretion of hCG, progesterone, and interleukin-6 (IL-6) by human term trophoblast cells in culture. Placentae were obtained from cesarean sections following uncomplicated pregnancies and used immediately after delivery. Leptin, hCG, progesterone, and IL-6 were measured by ELISA, RIA, and immunoradiometric assay in the cultured media of trophoblast cells cultured for 48 and 96 h. Leptin mRNA expression in these cultures was determined by reverse transcription-polymerase chain reaction. Recombinant human leptin added to primary cultures of human term placental trophoblast cells showed a stimulatory effect on hCG and IL-6 secretion and an inhibitory effect on progesterone secretion. Primary cultures of term trophoblast cells expressed leptin mRNA. All these findings suggest a role for leptin in human placental endocrine function. PMID- 12533411 TI - Protein inhibitor of nitric oxide synthase (NOS) and the N-methyl-D-aspartate receptor are expressed in the rat and mouse penile nerves and colocalize with penile neuronal NOS. AB - Nitrergic neurotransmission triggering penile erection is mediated by nitric oxide (NO) synthesized in the cavernosal nerves of the penis by penile neuronal NO synthase (PnNOS). In the central nervous system, nNOS is activated by the N methyl-D-aspartate receptor (NMDAR) and, presumably, is inhibited by the protein inhibitor of NOS (PIN). The PnNOS and NMDAR are expressed in the penis, and PnNOS has been localized in penile nerves. Both proteins colocalize with PIN in the hypothalamus and the spinal cord involved in the control of erection. The present study aimed to elucidate the relationship between PnNOS, PIN, and NMDAR in the penis. It was found that in the rat, PIN was expressed in the pelvic ganglion and the cavernosal nerve, and penile PIN cDNA was cloned, sequenced, and expressed. Immunohistochemistry localized PIN to the cavernosal and dorsal nerve of the penis, whereas NMDAR was not detected in the latter. Dual-fluorescence labeling showed that PnNOS colocalized with PIN in both nerves but with NMDAR only in the cavernosal nerve. Aging did not affect the mRNA levels of PnNOS, nNOS, NMDAR, and PIN. Both PIN and NMDAR were detected in penile nerves of the wild-type and nNOS( /-) mouse. The PIN protein did not inhibit or bind NOS in penile extracts, and in vivo, PIN cDNA reduced the erectile response to electrical field stimulation. In conclusion, PIN and NMDAR colocalize with PnNOS in penile nerves, but the functional significance of these protein interactions for penile erection remains to be elucidated. PMID- 12533413 TI - Messenger RNA (mRNA) expression for the antimicrobial peptides beta-defensin-1 and beta-defensin-2 in the male rat reproductive tract: beta-defensin-1 mRNA in initial segment and caput epididymidis is regulated by androgens and not bacterial lipopolysaccharides. AB - Mechanisms of antimicrobial protection in male reproductive organs are poorly understood. Defensins are antimicrobial peptides produced by many epithelial tissues. The goals of the present study were 1). to test the hypothesis that adult rat male reproductive organs express mRNA for rat beta-defensin (RBD)-1 and RBD-2, 2). to examine if defensin mRNA expression in the testis and epididymis is induced by bacterial lipopolysaccharides (LPS), and 3). to investigate the effects of androgens on defensin mRNA expression in the epididymis. Total RNA from reproductive organs was analyzed by relative reverse transcription polymerase chain reaction analysis. RBD-1 mRNA was detected in the testis. All segments of epididymis expressed equal levels of RBD-1 mRNA with higher expression than in the testis, whereas accessory sex glands showed expression equal to that in the testis. Expression of RBD-2 mRNA was primarily restricted to the penis. Effects of inflammation on defensin mRNA expression were examined in rats administered a unilateral injection of LPS from Pseudomonas aeruginosa or Escherichia coli. Expression of RBD-1 mRNA in the testis and epididymis was unaffected by LPS. To test the hypothesis that circulating androgens regulate RBD 1 mRNA expression in the epididymis, rats were subjected to bilateral orchiectomy (orch) or to orch plus a 3.5-cm implant containing testosterone. Expression of RBD-1 mRNA in the initial segment and caput was unchanged following 1-day orch but showed androgen-sensitive expression after 5 and 15 days. Expression of RBD-1 mRNA in corpus and cauda was not affected by orch. Results of this study suggest that RBD-1 may play an antimicrobial role in the testis and epididymis. PMID- 12533412 TI - Is the cadherin/catenin complex a functional unit of cell-cell actin-based adherens junctions in the rat testis? AB - Much controversy exists regarding the presence of the cadherin/catenin complex and its intracellular attachment site in the testis, which is the functional unit for actin-based cell-cell adherens junctions (AJs) in multiple epithelia. Furthermore, whether germ and Sertoli cells are equipped with the necessary AJ associated signaling molecules to regulate this cadherin/catenin complex during spermatogenesis is not known. In the present study, it was shown that both Sertoli and germ cells indeed express N-cadherin, E-cadherin, alpha-catenin, beta catenin, and p120(ctn) by semiquantitative reverse transcription-polymerase chain reaction and immunoblotting. Furthermore, the assembly of AJs between Sertoli and germ cells was associated with a transient induction in the steady-state mRNA and protein levels of cadherins and catenins. These analyses reveal, to our knowledge for the first time, that the testis may indeed be using the cadherin/catenin complex as one of the functional units to regulate AJ dynamics between Sertoli and germ cells in addition to alpha(6)beta(1) integrin and the nectin/afadin complex. To further confirm the existence of such a complex between Sertoli and germ cells, immunoprecipitation experiments were performed using Sertoli-germ cell lysates during AJ assembly. An anti-N-cadherin antibody can pull out beta catenin, whereas N-cadherin can also be pulled out using an anti-beta-catenin antibody. To further expand and validate these in vitro biochemical studies, immunofluorescent histochemistry was performed, which colocalized N-cadherin and beta-catenin to the same site of Sertoli-Sertoli and Sertoli-germ cell AJs, possibly ectoplasmic specializations near the basal compartment, at the lower third of the seminiferous epithelium in vivo as well as between Sertoli cells cultured in vitro. Furthermore, studies by cross-linking using dithiobis(succinimidylpropionate) confirmed that the cadherin/catenin complex between Sertoli cells as well as between Sertoli and germ cells indeed structurally linked to actin but not to vimentin (an intermediate filament protein) or to tubulin (a microtubule protein). These results thus unequivocally demonstrate that the cadherin/catenin complex, which can be up-regulated by testosterone, is indeed present between Sertoli and germ cells and is used for the assembly of functional AJs. PMID- 12533414 TI - Role of arachidonic acid and protein kinase C during maturation-inducing hormone dependent meiotic resumption and ovulation in ovarian follicles of Atlantic croaker. AB - The roles of arachidonic acid (AA) and protein kinase C (PKC) during in vitro maturation-inducing hormone (MIH)-dependent meiotic resumption (maturation) and ovulation were studied in ovarian follicles of Atlantic croaker (Micropogonias undulatus). The requirement for cyclooxygenase (COX) metabolites of AA was examined using a nonspecific COX inhibitor, indomethacin (IM), as well as two COX products, prostaglandin (PG) F(2alpha) and PGE(2), whereas the role of lipoxygenase (LOX) was investigated using a specific LOX inhibitor, nordihydroguaiaretic acid (NDGA). The involvement of PKC was examined using phorbol 12-myristate 13-acetate (PMA), a PKC activator, as well as GF109203X (GF), a specific inhibitor of PKC and 1-(5-isoquinolinesulfonyl)-2 methylpiperazine (H7), nonspecific inhibitor of protein kinases. Genomic mechanisms were examined with the transcription-inhibitor actinomycin D (ActD) and the functionality of heterologous (oocyte-granulosa) gap junctions (GJ) with a dye transfer assay. The AA (100 microM) and PGF(2alpha) (5 microM) did not induce maturation, and NDGA (10 microM) did not affect MIH-dependent maturation. However, IM (100 microM) partially inhibited MIH-dependent maturation. Conversely, AA and both PGs induced, and IM and NDGA inhibited, MIH-dependent ovulation in matured follicles. The PMA (1 microg/ml) did not induce maturation but caused ovulation in matured follicles, whereas PKC inhibitors (GF, 5 microM; H7, 50 microM) did not affect MIH-dependent maturation but inhibited MIH- and PMA dependent ovulation. The PMA-dependent ovulation was inhibited by IM but not by NDGA. In addition, ActD (5 microM) blocked MIH-dependent, but not PMA-dependent, ovulation, and PGF(2alpha) restored MIH-dependent ovulation in ActD-blocked follicles. The AA and PGs did not induce, and GF did not inhibit, MIH-dependent heterologous GJ uncoupling. In conclusion, AA and PKC mediate MIH-dependent ovulation but not meiotic resumption or heterologous GJ uncoupling in croaker follicles, but a permissive role of COX products of AA during maturation is possible. A novel model of MIH-dependent ovulation is proposed in which 1). LOX and COX metabolites of AA are both required for ovulation, but at upstream and downstream sites of the pathway, respectively, relative to PKC, and 2). PKC is downstream of genomic activation. PMID- 12533415 TI - Associated and independent comparisons between the two largest follicles preceding follicle deviation in cattle. AB - Follicle diameters and concentrations of follicular fluid factors were studied in the two largest follicles (F1 and F2) using F1 diameters in increments of 0.2 mm (equivalent to 4 h intervals) and extending from 7.4 to 8.4 mm (12 heifers in each of 6 groups). Changes were compared between follicles using the F2 associated with each F1-diameter group. Diameter deviation began in the 8.2-mm group as indicated by a greater (P < 0.05) diameter difference between F1 and F2 in the 8.4-mm group than in the 8.2-mm group. In the 8.0-mm group, estradiol concentrations began to increase (P < 0.05) differentially in F1 versus F2, and free insulin-like growth factor-1 (IGF-1) began to decrease differentially in F2 (P < 0.06). Combined for F1 and the associated F2, activin-A concentrations increased (P < 0.05) between the 7.6- and 8.2-mm groups and then decreased (P < 0.05). Results supported the hypothesis that estradiol and free IGF-1 concentrations simultaneously become higher in F1 than in the associated F2 by the beginning of diameter deviation. Results did not support the hypothesis that a transient elevation in activin-A is present in F1 but not in the associated F2 at the beginning of the estradiol and IGF-1 changes; instead, a mean transient elevation in activin-A occurred at this time only when data for the two follicles were combined. Comparisons between F1 and F2 also were made by independently grouping F2 and using diameter groups at 0.2-mm increments for F2 as well as for F1. In the diameter groups common to F1 and F2 (7.4, 7.6, 7.8, and 8.0 mm) there was a group effect (P < 0.003) for estradiol involving an increase (P < 0.05) beginning at the 7.6-mm group averaged over F1 and F2. For free IGF-1 concentrations, a fluctuation (a significant increase followed by a significant decrease) occurred independently in F1 between the 7.4- to 7.8-mm groups and independently in F2 between the 7.0- to 7.4-mm groups. PMID- 12533416 TI - Changes in the expression of tumor necrosis factor (TNF) alpha, TNFalpha receptor (TNFR) 2, and TNFR-associated factor 2 in granulosa cells during atresia in pig ovaries. AB - Tumor necrosis factor (TNF) alpha can induce both cell death and cell proliferation and exerts its effects by binding to either TNF receptor (TNFR) 1 or 2. When TNFalpha-bound TNFR2 interacts with TNFR-associated factor 2 (TRAF2), expression of survival/antiapoptotic genes is up-regulated. In the present study we determined the changes in localization of TNFalpha and TRAF2 and their mRNAs and the expression of TNFR2 in granulosa cells during follicular atresia in pig ovaries. In healthy follicles, intense signals for TNFalpha and TRAF2 and their mRNAs were demonstrated in the outer zone of the granulosa layer, where many proliferating cells and no apoptotic cells were observed. In atretic follicles, decreased or trace staining for TRAF2 and its mRNA and decreased expression of TNFR2 were observed in the granulosa layer, where many apoptotic cells were seen. These findings suggested that TNFalpha acts as a survival factor in granulosa cells during follicular atresia in pig ovaries. PMID- 12533417 TI - Cervical dilatation related to uterine electromyographic activity and endocrinological changes during prostaglandin F(2alpha)-induced parturition in cows. AB - The temporal relationship between changes in cervical dilatation, uterine electromyographic (EMG) activity, and maternal plasma concentrations of estradiol 17beta (E(2)), progesterone (P(4)), and 13,14-dihydro-15-keto-prostaglandin F(2alpha) (PGFM), was investigated in six parturient cows. Calving was induced with a single injection of a synthetic analogue of prostaglandin F(2alpha) (PG) on Day 274 of gestation. Cervical dilatation was measured continuously by measuring the transit time between two implanted ultrasound crystals while at the same time uterine EMG activity was measured through two silver electrodes sutured on the myometrial surface until the expulsive stage of calving had been reached. In blood samples collected at 4-h intervals, starting at the moment of PG injection, the mean plasma E(2) concentration gradually increased and was significantly elevated at 28 h after PG injection. At 4 h after PG treatment, the mean P(4) concentration had dropped significantly and continued to decrease until a value of around 1 ng/ml was reached, where it stayed until the onset of expulsion. Mean plasma PGFM concentrations increased steadily after PG injection, reaching significantly elevated concentrations at 20 h after treatment. In the five cows that delivered calves in anterior positions, uterine EMG activity, expressed as root mean square (RMS in microV), started to increase at a mean interval (+/- SD) of 13.1 +/- 3.7 h following PG treatment. The increase in EMG activity was significantly correlated with changes in plasma PGFM concentrations. In these cows, dilatation of the caudal cervix started after a mean (+/- SD) interval of 28.5 +/- 1.5 h following PG treatment and dilatation progressed at a mean (+/- SD) rate of 2.25 +/- 0.24 cm/h. In one cow with a calf in the posterior position, uterine EMG activity and dilatation started at 15.8 h and 31.8 h, respectively, after induction of calving. We conclude that a predictable sequence of physiological changes occurs around induction of calving, which allows specific timing of future studies on cellular and biochemical changes within the cervix during parturition. PMID- 12533419 TI - Temporally controlled site-specific mutagenesis in the germ cell lineage of the mouse testis. AB - We have obtained a PrP-Cre-ER(T) transgenic mouse line (28.8) that selectively expresses in testis the tamoxifen-inducible Cre-ER(T) recombinase under the control of a mouse Prion protein (PrP) promoter-containing genomic fragment. Cre ER(T) is expressed in spermatogonia and spermatocytes, but not in Sertoli and Leydig cells. We also established reporter PrP-L-EGFP-L transgenic mice harboring a LoxP-flanked enhanced green fluorescent protein (EGFP) Cre reporter cassette under the control of the same PrP promoter-containing genomic fragment that exhibits prominent EGFP expression in brain and testis. Using the PrP-L-EGFP-L as well as other Cre-reporter mice, we demonstrate that tamoxifen administration efficiently and selectively induces Cre-mediated recombination in the germ cell lineage. The established PrP-Cre-ER(T) line should provide a valuable tool for studying functions of germ cell-expressed genes involved in spermatogenesis. PMID- 12533418 TI - Novel RING finger protein OIP1 binds to conserved amino acid repeats in sperm tail protein ODF1. AB - Outer dense fibers (ODFs) and the fibrous sheath (FS) are unique structures of the mammalian sperm tail. Recently, progress has been made in the molecular cloning of ODF and FS proteins, and because of this, questions addressing the morphogenesis and underlying protein network that make up sperm tail structures and their function can now be addressed. Using the N-terminal leucine zipper motif of the major ODF protein ODF1, we had previously isolated interacting proteins Odf2, Spag4, and Spag5. We report here a yeast two-hybrid strategy to isolate a novel rat testicular protein, OIP1, that binds to the evolutionarily conserved Cys-Gly-Pro repeats in the C-terminus of ODF1. OIP1 is expressed in round spermatids as well as in spermatocytes and several somatic tissues, albeit at a lower level. No expression was detectable in epididymis, heart, and smooth muscle. OIP1 protein localizes to the sperm tail in a pattern expected for an ODF1-interacting protein. OIP1 belongs to the family of RING finger proteins of the H2 subclass. Deletion of the putative RING motif significantly decreased binding to ODF1. Genomic analysis of rat Oip1 and Oip1 homologs indicates that Oip1 is highly conserved. Oip1 is subject to differential splicing and alternative polyadenylation events. It is interesting that Oip1 mRNAs have been reported that lack the exon encoding the putative RING finger. PMID- 12533420 TI - DMRT1 is upregulated in the gonads during female-to-male sex reversal in ZW chicken embryos. AB - Sex in birds is chomosomally based (ZZ male, ZW female), but the mechanism underlying sex determination remains unknown. An unresolved question is whether Z gene dosage plays a role in avian sex determination. DMRT1 is an avian Z-linked gene that shows higher expression in male gonads during embryogenesis and has been proposed as a putative testis-determining gene in birds. The Z-linkage of this gene makes it an ideal candidate for testing the question of gene dosage in avian testis determination. A higher level of DMRT1 expression in male (ZZ) versus female (ZW) embryonic gonads may reflect the presence of two Z-linked copies in the male, or it may be due to specific and active upregulation of DMRT1 during testis formation. A functional interventionist strategy was used to distinguish between these two possibilities. DMRT1 expression was analyzed in chicken embryos during experimentally induced female-to-male sex reversal, using the aromatase enzyme inhibitor fadrozole. DMRT1 expression was analyzed by whole mount in situ hybridization and reverse transcription polymerase chain reaction (for mRNA) and indirect immunofluorescence (for protein). Female-to-male sex reversed embryos (genetically ZW) showed elevated levels of DMRT1 expression similar to those of normal males (with two copies of the Z chromosome). Elevated levels of DMRT1 are therefore associated with testis development, both in normal males (ZZ) and in sex-reversed females (ZW). SOX9 expression was also activated during female-to-male sex reversal but appeared delayed relative to DMRT1 upregulation. These results show that testis development does not require two Z linked copies of DMRT1, but it does involve active upregulation of the gene. Higher levels of DMRT1 expression during testis differentiation therefore do not simply reflect a gene dosage difference between the two sexes but imply active involvement in male development. PMID- 12533421 TI - Handmade somatic cell cloning in cattle: analysis of factors contributing to high efficiency in vitro. AB - Widespread application of somatic cell cloning has been hampered by biological and technical problems, which include complicated and time-consuming procedures requiring skilled labor. Recently, zona-free techniques have been published with limited or no requirement for micromanipulators. The purpose of the present work was to optimize certain steps of the micromanipulator-free (i.e., handmade) procedure, to analyze the morphology of the developing blastocysts, and to explain factors involved in the high efficiencies observed. Optimization of the procedure included selection of the appropriate medium for enucleation, orientation of pairs at fusion, timing of fusion, and culture conditions. As a result of these improved steps, in vitro efficiency as measured by blastocysts per reconstructed embryo and blastocysts per working hour was among the highest described so far. The cattle serum used in our experiments was superior to other protein sources for in vitro embryo development. One possible explanation of this effect is the considerable mitogenic activity of the cattle serum compared with that of commercially available fetal calf serum. Morphological analysis of blastocysts by inverted microscopy, inner cell mass-trophoblast differential staining, and transmission electron microscopy revealed high average quality. A high initial pregnancy rate was achieved after the transfer of single blastocysts derived by aggregation of two nuclear transfer embryos into recipients. The improved handmade somatic cell nuclear transfer method may become a useful technology as a simple, inexpensive, and efficient alternative to traditional somatic cell nuclear transfer. PMID- 12533422 TI - Mouse placental prostaglandins are associated with uterine activation and the timing of birth. AB - We explored a potential mechanism linking placental prostaglandins (PGs) with a fall in plasma progesterone and increased expression of uterine activation proteins in the mouse. PG endoperoxide H synthase 2 (PGHS-2) mRNA expression increased in placenta in late gestation in association with an 8-fold increase in PGF(2alpha) concentration, reaching a peak on Gestational Day (GD) 18. This peak coincided with the final descent in plasma progesterone and birth on GD 19.3 +/- 0.2. Implantation of a progesterone-releasing pellet in intact pregnant dams on GD 16 delayed birth at term until GD 20.9 +/- 0.4 and inhibited the GD 18 increase in placental PGF(2alpha) levels in conjunction with a delayed fall in plasma progesterone that reached its lowest level 1 day after term birth. The mRNA levels of uterine activation proteins, connexin-43 (CX-43), oxytocin receptor, PGF(2alpha) receptor (FP), and PGHS-2, and the concentration of uterine PGF(2alpha) all increased at normal term birth. At progesterone-delayed term birth on GD 19.3, even though tissue PGF(2alpha) concentrations were at the same high levels observed at normal term birth, CX-43 and FP mRNA levels were lower than those at normal term birth, thereby possibly contributing to the delay of birth. These data are consistent with the hypotheses that fetal placental PGs affect the timing of birth by hastening luteolysis, that uterine activation initiates labor, and that birth may be delayed by blocking or decreasing the expression of two of the uterine activation proteins. PMID- 12533423 TI - Quantification of prolactin (PRL) and PRL receptor messenger RNA in gilthead seabream (Sparus aurata) after treatment with estradiol-17beta. AB - Prolactin (PRL) in fish is considered to be an osmoregulatory hormone, although some studies suggest that it may influence the production of steroid hormones in the gonads. The objective of the present study was to establish if PRL is involved in reproduction of the gilthead seabream-a protandrous hermaphrodite. Adult and juvenile gilthead seabream received implants of estradiol-17beta (E(2)) for 1 wk during the breeding season, and the mRNA expressions of PRL and PRL receptor (sbPRLR) were determined. Northern blot analysis revealed a single pituitary PRL transcript, the expression of which was significantly reduced by E(2) treatment in adults but significantly increased in juvenile fish. In adult gonads, four sbPRLR transcripts of 1.1, 1.3, 1.9, and 2.8 kilobases were observed. A competitive reverse transcription-polymerase chain reaction was developed and used to determine how E(2) treatment alters expression of the gonadal sbPRLR gene. Seabream PRLR was detectable in all samples analyzed by this assay. Levels of sbPRLR mRNA increased significantly (50-fold) after E(2) treatment in adults, but a 24-fold decrease was measured in juveniles. Immunohistochemistry using specific polyclonal antibodies raised against an oligopeptide from the extracellular domain of sbPRLR detected the receptor in spermatogonia and oocytes. Taken together, the preceding results suggest that in the seabream, PRL may act on both testis and ovary via its receptor and that the stage of maturity influences this process. The full characterization and relative importance of the different transcripts of sbPRLR in eliciting the action of PRL in the gonads remain to be elucidated. PMID- 12533424 TI - Effect of cell confluence on production of cloned mice using an inbred embryonic stem cell line. AB - Mice have been successfully cloned from both somatic cells and hybrid embryonic stem (ES) cells. Heterozygosity of the donor ES cell genome has been suggested as a crucial factor for long-term survival of cloned mice. In the present study, an inbred ES cell line, HM-1 (129/Ola), and a well-tested ES cell line, R1 (129/Sv x 129/Sv-CP), were used as donor cells to evaluate the developmental potential of nuclear transfer embryos. We found that ES cell confluence dramatically affects the developmental potential of reconstructed embryos. With the ES cell line HM-1 and 80-90% confluence, 49% of reconstructed embryos developed to the morula/blastocyst stage, 9% of these embryos developed to live pups when transferred to the surrogate mothers, and 5 of 18 live pups survived to adulthood. By contrast, at 60-70% confluence, only 22% of embryos developed to the morula/blastocyst stage, and after transfer, only a single fetus reached term. Consistent with previous reports, the nuclei of R1 ES cells were also shown to direct development to term, but no live pups were derived from cells at later passages (>20). Our results show that the developmental potential of reconstructed embryos is determined by both cell confluence and cell passage. These results also demonstrate that the inbred ES cell line, HM-1, can be used to produce viable cloned mice, although less efficiently than most heterozygous ES cell lines. PMID- 12533425 TI - Analyses of mitogen-activated protein kinase function in the maturation of porcine oocytes. AB - The function of mitogen-activated protein kinase (MAPK) during porcine oocyte maturation was examined by injecting oocytes with either mRNA or antisense RNA of porcine c-mos protein, an upstream kinase of MAPK. The RNAs were injected into the cytoplasm of porcine immature oocytes immediately after collection from ovaries, then the oocytes were cultured for maturation up to 48 h. The phosphorylation and activation of MAPK were observed at 6 h after injection of the c-mos mRNA injected-oocytes, whereas in control oocytes, MAPK activation was detected at 24 h of culture. The germinal vesicle breakdown (GVBD) rate at 24 h of culture was significantly higher in c-mos mRNA-injected oocytes than in control oocytes. In contrast, although injection of c-mos antisense RNA completely inhibited phosphorylation and activation of MAPK throughout the maturation period, the GVBD rate and its time course were the same in noninjected oocytes. The degree of maturation-promoting factor (MPF) activation was, however, very low in oocytes in the absence of MAPK activation. Most of those oocytes had both abnormal morphology and decondensed chromosomes at 48 h of culture. These results suggest that MAPK activation is not required for GVBD induction in porcine oocytes and that the major roles of MAPK during porcine oocyte maturation are to promote GVBD by increasing MPF activity and to arrest oocytes at the second metaphase. PMID- 12533426 TI - Role and gonadotrophic regulation of X-linked inhibitor of apoptosis protein expression during rat ovarian follicular development in vitro. AB - Although FSH up-regulates follicular cell X-linked inhibitor of apoptosis protein (XIAP) expression and suppresses apoptosis in vivo, if these events are coincidental or causally related remains to be investigated. The present study examined the role and gonadotrophic regulation of XIAP expression during follicular development in vitro. Follicles (160-210 microm) cultured for 0-6 days with FSH (100 ng/ml) showed significant growth, as evidenced by increases in follicular size, cell number, and DNA contents. Follicular XIAP content was low in the absence of FSH but was increased by the addition of gonadotropin. Apoptosis was evident in follicles cultured without FSH but was suppressed in the presence of gonadotropin. At low FSH concentration (5 ng/ml), adenoviral XIAP sense cDNA expression increased XIAP and DNA contents, reduced apoptosis, and enhanced follicular growth. Infection of the FSH-stimulated follicles with XIAP antisense elicited opposite responses. In primary granulosa cell cultures, FSH significantly increased XIAP content, inhibited apoptosis, and decreased cell number, a response potentiated by XIAP sense expression. In conclusion, the present studies demonstrated, to our knowledge for the first time, that XIAP plays an important role in the regulation of ovarian follicular development. In addition, a follicle culture system coupled to an adenoviral gene-manipulation procedure has been established and may prove to be a useful approach in assessing the role of specific genes in follicular development and atresia. PMID- 12533427 TI - Activin and follicle-stimulating hormone signaling are required for long-term culture of functionally differentiated primary granulosa cells from the chicken ovary. AB - Follicle-stimulating hormone, activin A, and transforming growth factor (TGF) alpha are important regulators of chicken granulosa cell (cGC) function. Hence, we aimed to test whether these growth factors are useful for establishing a suitable in vitro cell culture model system of primary cGC. Although cGC are easily isolated from distinct follicular stages, a long-term cGC culture system for in vitro studies has been unavailable. Here, we report a novel, long-term cell culture system that allows for cGC proliferation in vitro while maintaining the epithelial phenotype that granulosa cells exhibit in vivo. The cGC rapidly lose their epithelial morphology and acquire a mesenchymal or fibroblastoid phenotype when cultured in the absence of activin A. This process is strongly enhanced by TGFalpha, a well-known granulosa cell mitogen. However, FSH stimulates cGC proliferation without enhancing morphological changes and dedifferentiation. Interestingly, a combination of both activin A and FSH stimulates cGC proliferation and supports maintenance of differentiated epithelial morphology. Furthermore, activin A and FSH synergistically induce granulosa cell-specific differentiation markers such as inhibin alpha and chicken zona pellucida protein C, suggesting that cultured cGC resemble functionally differentiated granulosa cells. Our data demonstrate that activin signaling is necessary to sustain a morphologically differentiated phenotype of cGC in vitro. The results also suggest a pivotal importance of activin signaling for granulosa cell function in vivo. PMID- 12533428 TI - DNA double-strand breaks and gamma-H2AX signaling in the testis. AB - Within minutes of the induction of DNA double-strand breaks in somatic cells, histone H2AX becomes phosphorylated at serine 139 and forms gamma-H2AX foci at the sites of damage. These foci then play a role in recruiting DNA repair and damage-response factors and changing chromatin structure to accurately repair the damaged DNA. These gamma-H2AX foci appear in response to irradiation and genotoxic stress and during V(D)J recombination and meiotic recombination. Independent of irradiation, gamma-H2AX occurs in all intermediate and B spermatogonia and in preleptotene to zygotene spermatocytes. Type A spermatogonia and round spermatids do not exhibit gamma-H2AX foci but show homogeneous nuclear gamma-H2AX staining, whereas in pachytene spermatocytes gamma-H2AX is only present in the sex vesicle. In response to ionizing radiation, gamma-H2AX foci are generated in spermatogonia, spermatocytes, and round spermatids. In irradiated spermatogonia, gamma-H2AX interacts with p53, which induces spermatogonial apoptosis. These events are independent of the DNA-dependent protein kinase (DNA-PK). Irradiation-independent nuclear gamma-H2AX staining in leptotene spermatocytes demonstrates a function for gamma-H2AX during meiosis. gamma-H2AX staining in intermediate and B spermatogonia, preleptotene spermatocytes, and sex vesicles and round spermatids, however, indicates that the function of H2AX phosphorylation during spermatogenesis is not restricted to the formation of gamma-H2AX foci at DNA double-strand breaks. PMID- 12533429 TI - Expression of wilms' tumor gene and protein localization during ovarian formation and follicular development in sheep. AB - Wilms' tumor protein (WT1) is a transcriptional repressor essential for the development of mammalian kidneys and gonads. To gain insight into possible roles of WT1 in ovarian formation and follicular function, we studied patterns of mRNA and protein localization throughout fetal gonadal development and in ovaries of 4 wk-old and adult sheep. At Day 24 after conception, strong expression of WT1 mRNA and protein was observed in the coelomic epithelial region of the mesonephros where the gonad was forming. By Day 30, expression was observed in the surface epithelium and in many mesenchymal and endothelial cells of the gonad. Epithelial cells continued to express WT1 throughout gonadal development, as did pregranulosa cells during the process of follicular formation. However, WT1 expression was not observed in germ cells. During follicular growth, granulosa cells expressed WT1 from the type 1 (primordial) to the type 4 stages, but thereafter expression was reduced in type 5 (antral) follicles, consistent with the differentiation of granulosa cells into steroid-producing cells. The possible progenitor cells for the theca interna (i.e., the cell streams in the ovarian interstitium) expressed WT1 heterogeneously. However, differentiated theca cells in antral follicles did not express WT1. Strong expression of WT1 was observed during gonadal development, which is consistent with a role for WT1 in ovarian and follicular formation in the ewe. WT1 was identified in many cells of the neonatal and adult ovaries, including granulosa cells, suggesting that this factor is important for preantral follicular growth. However, the decline in WT1 expression in antral follicles suggests that WT1 may prevent premature differentiation of somatic cells of the follicle during early follicular growth. PMID- 12533430 TI - 5' hox genes and meis 1, a hox-DNA binding cofactor, are expressed in the adult mouse epididymis. AB - Hox genes determine the formation of segmented structures during development. The epididymis shows a segmented organization in its structure and function beyond embryogenesis. This study examined the adult mouse epididymis and vas deferens for expression of 5' hox genes and a hox-DNA binding cofactor. Reverse transcriptase-polymerase chain reaction (RT-PCR) showed the expression of hoxa-9, hoxa-10, hoxa-11, hoxd-9, and hoxd-10 in all regions including the vas deferens. Semiquantitative RT-PCR revealed highest mRNA levels for hoxa-11 in the distal part of the epididymis and vas deferens, and this was confirmed by Northern blot analysis. To determine protein presence an antibody raised against a peptide N terminal to the homeodomain of hoxa-11 was produced in rabbits. The antibody recognized a band of approximately 37-39 kDa in Western blot analysis. Immunohistochemistry indicated the presence of hoxa-11 in the nuclei of the epithelial cells with some staining in the cytoplasm. Staining was also detected in nuclei of interstitial cells throughout the entire organ and the vas deferens. A DNA binding cofactor for hoxa-11, Meis 1, was investigated for its presence in the epididymis. Semiquantitative RT-PCR identified both transcripts for Meis 1 (Meis 1a and Meis 1b) in all regions. Protein presence was confirmed by Western blot analysis, and this detected one band of approximately 53-55 kDa. Immunohistochemistry localized Meis 1 in the nuclei of interstitial cells throughout the entire organ and the vas deferens. Our study provides preliminary data from which we suggest the involvement of homeodomain transcription factors in the maintenance of segmental function of the adult epididymis and vas deferens. PMID- 12533431 TI - MuERV-L is one of the earliest transcribed genes in mouse one-cell embryos. AB - The expression pattern and function of the murine endogenous retrovirus-like (MuERV-L) gene in mouse preimplantation embryos was investigated. MuERV-L was rapidly transcribed from the beginning of S phase (8 h after fertilization) in the first cell cycle. MuERV-L expression was completely repressed when transcription from the zygotic genome was inhibited by alpha-amanitin. These results reveal that MuERV-L is transcribed from the zygotic genome and that it is expressed earlier than any other genes previously reported. In addition, MuERV-L was expressed even when the first round of DNA synthesis was inhibited by aphidicolin, suggesting that its expression is controlled by the zygotic clock. The function of MuERV-L in the development of mouse embryos was also examined using antisense oligonucleotides. The developmental competence of embryos was markedly suppressed after the 4-cell stage when they were treated with antisense oligonucleotides. This result suggests that MuERV-L plays an important role in the development of mouse embryos at the early preimplantation stage. PMID- 12533432 TI - Fate of postacrosomal perinuclear theca recognized by monoclonal antibody MN13 after sperm head microinjection and its role in oocyte activation in mice. AB - Monoclonal antibody (mAb) MN13 labels mouse sperm head postacrosomal perinuclear theca (PT), which is possibly involved in oocyte activation during fertilization. The antigenic site is expressed after mild sonication followed by treatment with dithiothreitol (DTT) or heat (45 degrees C), and is visible as a thick band in the postacrosomal region. The presence of protease inhibitors in the sonication medium suppresses the exposure of MN13 epitope (MN13p), suggesting the involvement of a proteolytic reaction in this process. Spermatozoa do not express MN13p after the induction of acrosome exocytosis by Ca(2+) ionophore, zona binding, or during zona penetration, a strategy that ensures safe delivery of postacrosomal PT proteins to oocytes after fusion. MN13 labeling was not detectable during fertilization by zona-free in vitro fertilization, suggesting that the antigenic site does not react with proteolytic enzymes during sperm oocyte fusion and the antibody does not recognize the nascent epitope. Microinjection of sperm heads prepared by sonication and DTT treatment led to the activation of metaphase II oocytes. The oocyte activating function of such sperm heads was significantly diminished after labeling with MN13 prior to intracytoplasmic sperm injection (ICSI), but labeling with antiequatorin antibody MN9 activated oocytes with a frequency similar to that of unlabeled sperm heads. The sperm heads in inactive oocytes formed premature chromosome condensations (PCCs), which were invested by independent metaphase-like spindles. These observations indicate that the postacrosomal PT recognized by mAb MN13 is involved in oocyte activation. MN13p is dissociated from sperm heads during the early stages of decondensation after ICSI. In activated oocytes, MN13-labeled fine granules were redistributed in the midzone spindle region, whereas in inactive oocytes they formed a ring around the polar regions of the metaphase II and PCC spindles. PMID- 12533433 TI - Real-time observations of individual macaque sperm undergoing tight binding and the acrosome reaction on the zona pellucida. AB - Changes in binding affinity, acrosomal status, and motility of living sperm on the zona pellucida were for the first time in any mammalian species directly observed and analyzed with video microscopy. A single zona was air-dried and rehydrated on a microscope slide, and a coverslip supported by glass beads was added. Capacitated sperm were added together with Alexa-SBTI, a probe for acrosin that can detect the acrosome reaction. The heads of loosely attached sperm oscillated on the zona and the flagella beat symmetrically with a sigmoid-shaped waveform. Tight binding was observed after 16 sec as the sperm head became fixed in place on the zona. The shape of the flagellar beat simultaneously shifted to a more rigid, C-shaped waveform. The first signs of the acrosome reaction were detected within 11 sec of tight binding. Rapid flushing removed approximately 65% of sperm that were loosely attached but only 2% of those that were tightly bound. In the 2 min following the onset of tight binding, the lateral displacement of the flagellum increased by approximately 30% and the beat frequency decreased by 25%. Lignosulfonic acid (LSA) inhibited loose sperm attachment and the development of tight binding. LSA had no effect on the time of the acrosome reaction following tight binding or on changes in motility that followed tight binding. These data suggest that LSA affects the initial attachment or docking of sperm to the zona, a step that may align or recruit one or more specific zona receptors to be responsible for mediating the acrosome reaction. PMID- 12533434 TI - Progesterone-modulated induction of apoptosis by interferon-tau in cultured epithelial cells of bovine endometrium. AB - Interferon-tau (IFN-tau) is produced by the trophoblast prior to implantation in ruminants. It is involved in maternal recognition of pregnancy, and is a pleiotropic molecule that can alter the synthesis of endometrial proteins and inhibit proliferation of some cells. We have observed that IFN-tau reduces the DNA content in cultures of bovine endometrial epithelial cells; therefore, the objective of this study was to determine whether IFN-tau would induce apoptosis in bovine endometrial cells. Epithelial cells were prepared, cultured to confluence, and then incubated for 24 or 48 h in the presence or absence of 10 ng/ml progesterone, 100 ng/ml IFN-tau, or 10 microg/ml cycloheximide (CHX; an apoptosis inducer used as a positive control). Cells undergoing apoptosis exhibit such characteristics as the appearance of apoptotic bodies and DNA fragmentation. The incidence of apoptosis was assessed by using TUNEL, DNA fragmentation analysis, and Western blot analysis of Bax-alpha protein expression. The results showed that IFN-tau and CHX significantly increased the percentage of cells with apoptotic nuclei (33.6% and 44.8%, respectively) compared with controls (11.7%; P < 0.05). Progesterone treatment of the cells significantly inhibited the ability of IFN-tau to induce apoptosis (14.6%) compared with IFN-tau alone (33.6%; P < 0.05). DNA fragmentation analysis showed that INF-tau and CHX treatment resulted in an increase in the appearance of DNA laddering compared with that in untreated control cultures. Western blot analysis showed that IFN-tau and CHX treatment resulted in a greater expression of the proapoptotic protein Bax-alpha compared with that in control cultures. These data demonstrate that IFN-tau can induce apoptosis in bovine uterine epithelial cells and that this effect is modulated by progesterone. We speculate that IFN-tau might play a critical role in the remodeling of the endometrium around the time of implantation. PMID- 12533435 TI - Mice that express enzymatically inactive cathepsin L exhibit abnormal spermatogenesis. AB - The finding of large, stage-specific changes in secretion of procathepsin L by rat Sertoli cells has led to the hypothesis that this proenzyme promotes the survival, replication, or differentiation of spermatogenic cells. Experiments described herein used a mouse model to test this hypothesis. To prove that mice are appropriate for this purpose, we first demonstrate that mature mouse Sertoli cells express cathepsin L mRNA in the same stage-specific manner as rat Sertoli cells and they also secrete procathepsin L. To test whether catalytically active cathepsin L is required for normal spermatogenesis, we examined the testes of 110 to 120-day-old furless mice, which express catalytically inactive cathepsin L. Morphologic examination of testes of furless mice revealed both normal and atrophic seminiferous tubules. Enumeration of atrophic tubules in furless and control mice demonstrates that lack of functional cathepsin L results in a 12 fold increase in seminiferous tubule atrophy. To determine whether lack of functional cathepsin L affects the production of male germ cells in apparently normal, nonatrophic tubules, we compared numbers in control and furless mice of preleptotene spermatocytes, pachytene spermatocytes, and round spermatids per Sertoli cell. Results demonstrate that the lack of functional cathepsin L causes a 16% reduction in formation of preleptotene spermatocytes and a 25% reduction in differentiation of these cells into pachytene spermatocyte. These results suggest that procathepsin L either directly or indirectly has two distinct functions in the testis. This proenzyme prevents atrophy of seminiferous tubules and promotes the formation of preleptotene spermatocytes and the differentiation of these meiotic cells into pachytene spermatocytes. PMID- 12533436 TI - Differential ubiquitination of stallion sperm proteins: possible implications for infertility and reproductive seasonality. AB - Antibodies against ubiquitin, a universal proteolytic marker, show increased cross-reactivity with defective spermatozoa in men and bulls. We investigated sperm ubiquitination in the stallion, a seasonally polyestrous mammal. Immunofluorescence and immunoelectron microscopy demonstrated that anti-ubiquitin antibodies bind to the surface of both membrane-intact and aldehyde-fixed spermatozoa. Cross-reactivity to the ubiquitin-conjugating enzyme E2 was also detected in sperm. Immunohistochemistry showed that ubiquitinated spermatozoa were first detected in the caput epididymis, coincident with a strong accumulation of ubiquitin and ubiquitin C-terminal hydrolase, protein gene product 9.5, in the apical stereocilia of the epididymal epithelium. Testicular spermatozoa did not display significant ubiquitin cross-reactivity. Similarly, lesser accumulation of ubiquitin cross-reactive substrates was identified in the accessory sex glands. Semen samples were collected from three fertile stallions and one subfertile stallion between December and February and probed for ubiquitin by flow cytometry and immunoblotting. Flow cytometric analysis showed that sperm from the subfertile stallion had higher ubiquitin levels than sperm from the other three stallions. In addition, immunoblot analysis of sperm proteins from the subfertile stallion showed two unique ubiquitin cross-reactive bands that were not present in sperm extracts from the three fertile stallions. To screen for a possible role for ubiquitin in seasonal changes in sperm production, semen samples from two fertile stallions were collected in March, June, September, and December and subjected to a flow cytometric ubiquitin assay. The lowest levels of ubiquitin-labeled sperm were found in March, approximately coincident with the onset of the natural horse breeding season. A progressive increase in sperm ubiquitin levels was found during summer and fall, with a peak in December. These data suggest that stallion sperm are differentially ubiquitinated during epididymal maturation and that this ubiquitination may reflect changes in sperm numbers and semen quality. The association between changes in sperm ubiquitination and seasonal changes in sperm production will be subjected to further studies in a larger cohort of animals. PMID- 12533437 TI - Testis-specific expression of rat mitochondrial glycerol-3-phosphate dehydrogenase in haploid male germ cells. AB - Mitochondrial glycerol-3-phosphate dehydrogenase (mGPDH) is regulated by multiple promoters in a tissue-specific manner. We characterized the testis-specific promoter C of the mGPDH gene and investigated the cellular localization of mGPDH within the testis. Electrophoretic mobility shift experiments identified a cAMP response element (CRE) site at -57 that was active in the testis. An in vitro translated CRE modulator (CREM) protein was able to bind this CRE site, and an anti-CREM antibody interfered with this complex. Ectopic expression of the testis specific transcriptional activator CREMtau and protein kinase A in human hepatocarcinoma HepG2 cells activated a promoter C-driven luciferase construct in transient transfection experiments. Furthermore, mGPDH expression was undetectable in testis of CREM-deficient mice. The cellular localization of mGPDH expression and translation in adult rat testis was determined by in situ hybridization and immunohistochemistry techniques. The mGPDH transcripts were detected solely in postmeiotic germ cells. Expression of mGPDH was restricted from round spermatids to early elongating spermatids. The mGPDH protein was delayed in postmeiotic germ cells, restricted from late elongating spermatids to mature spermatids. Our results indicate that rat mGPDH is expressed by a testis specific promoter from haploid male germ cells in a stage-specific manner. PMID- 12533438 TI - Role of platelet-derived growth factor in vascular remodeling during pulmonary hypertension in the ovine fetus. AB - Platelet-derived growth factor (PDGF) is a potent smooth muscle cell mitogen that may contribute to smooth muscle hyperplasia during the development of chronic pulmonary hypertension (PH). We studied changes in PDGFalpha- and beta-receptor and ligand expression in lambs with chronic intrauterine PH induced by partial ligation of the ductus arteriosus (DA) at gestational age 124-128 days (term = 147 days). Western blot analysis performed on whole lung homogenates from PH animals after 8 days of DA ligation showed a twofold increase in PDGFalpha- and beta-receptor proteins compared with age-matched controls (P < 0.05). Lung PDGF-A and -B mRNA expression did not differ between PH and control animals. We treated PH animals with NX1975, an aptamer that selectively inhibits PDGF-B, by infusion into the left pulmonary artery for 7 days after DA ligation. NX1975 reduced the development of muscular thickening of small pulmonary arteries by 47% (P < 0.05) and right ventricular hypertrophy (RVH) by 66% (P < 0.02). Lung PDGFalpha- and beta-receptor expression is increased in perinatal PH, and NX1975 reduces the increase in wall thickness of small pulmonary arteries and RVH in this model. We speculate that PDGF signaling contributes to structural vascular remodeling in perinatal PH and that selective PDGF inhibition may provide a novel therapeutic strategy for the treatment of chronic PH. PMID- 12533439 TI - Developmental differences in the shear stress-induced expression of endothelial NO synthase: changing role of AP-1. AB - Endothelial nitric oxide synthase (eNOS) mRNA and protein levels increase during late gestation and then decrease postnatally in sheep lung parenchyma. The increase in fluid shear stress at birth, resulting from increased pulmonary blood flow, is an important mediator of postnatal eNOS gene expression. Our objective was to identify factors stimulating eNOS expression in pulmonary arterial endothelial cells (PAEC) in response to shear stress and to determine if these factors are developmentally regulated. PAEC were isolated from fetal lambs and adult sheep. Transcriptional activity from a 1,600-bp eNOS promoter fragment increased in both fetal and adult PAEC exposed to 8 h of shear stress. Conversely, activity driven from an 840-bp promoter fragment containing a putative activator protein (AP)-1 binding site was increased only in fetal PAEC. This increase was completely abolished in an identical construct containing a mutant AP-1 sequence. The AP-1 protein c-Jun was localized to the cytosol in static adult PAEC and to the nucleus in static fetal PAEC. After shear, c-Jun was nuclear localized in both cell types. However, transcriptionally active phosphorylated c-Jun was elevated only in the nuclei of sheared fetal PAEC. Resting levels of eNOS and NO were 2- and 20-fold higher, respectively, in fetal cells. Shear increased eNOS and NO in both cell types: levels were approximately 2.5-fold higher in fetal PAEC. Phosphorylation of Akt and eNOS was evident in sheared fetal but not adult PAEC. We have therefore identified mechanisms of eNOS regulation at the transcriptional level and to be enzyme activation specific to the fetal pulmonary arterial circulation. PMID- 12533440 TI - Contributions of nitric oxide synthase isozymes to exhaled nitric oxide and hypoxic pulmonary vasoconstriction in rabbit lungs. AB - We investigated the source(s) for exhaled nitric oxide (NO) in isolated, perfused rabbits lungs by using isozyme-specific nitric oxide synthase (NOS) inhibitors and antibodies. Each inhibitor was studied under normoxia and hypoxia. Only nitro L-arginine methyl ester (L-NAME, a nonselective NOS inhibitor) reduced exhaled NO and increased hypoxic pulmonary vasoconstriction (HPV), in contrast to 1400W, an inhibitor of inducible NOS (iNOS), and 7-nitroindazole, an inhibitor of neuronal NOS (nNOS). Acetylcholine-mediated stimulation of vascular endothelial NOS (eNOS) increased exhaled NO and could only be inhibited by L-NAME. Selective inhibition of airway and alveolar epithelial NO production by nebulized L-NAME decreased exhaled NO and increased hypoxic pulmonary artery pressure. Immunohistochemistry demonstrated extensive staining for eNOS in the epithelia, vasculature, and lymphatic tissue. There was no staining for iNOS but moderate staining for nNOS in the ciliated cells of the epithelia, lymphoid tissue, and cartilage cells. Our findings show virtually all exhaled NO in the rabbit lung is produced by eNOS, which is present throughout the airways, alveoli, and vessels. Both vascular and epithelial-derived NO modulate HPV. PMID- 12533441 TI - Characterization of adenosine receptor(s) involved in adenosine-induced bronchoconstriction in an allergic mouse model. AB - We recently reported that adenosine caused bronchoconstriction and enhanced airway inflammation in an allergic mouse model. In this study, we further report the characterization of the subtype of adenosine receptor(s) involved in bronchoconstriction. 5'-(N-ethylcarboxamido)adenosine (NECA), a nonselective adenosine agonist, elicited bronchoconstriction in a dose-dependent manner. Little effects of N(6)-cyclopentyladenosine (A(1)-selective agonist) and 2-p-(2 carboxyethyl)phenethylamino-5'-N-ethylcarboxamidoadenosine (A(2A)-selective agonist) compared with NECA were observed in this model. 2-Chloro-N(6)-(3 iodobenzyl)-9-[5-(methylcarbamoyl)-beta-d-ribofuranosyl]adenosine, an A(3) selective receptor agonist, produced a dose-dependent bronchoconstrictor response, which was blocked by selective A(3) antagonist 2,3-diethyl-4,5-dipropyl 6-phenylpyridine-3-thiocarboxylate-5-carboxylate (MRS1523). However, MRS1523 only partially inhibited NECA-induced bronchoconstriction. Neither selective A(1) nor A(2A) antagonists affected NECA-induced bronchoconstriction. Enprofylline, a relatively selective A(2B) receptor antagonist, blocked partly NECA-induced bronchoconstriction. Furthermore, a combination of enprofylline and MRS1523 completely abolished NECA-induced bronchoconstrictor response. Using RT-PCR, we found that all four adenosine receptor subtypes are expressed in control lungs. Allergen sensitization and challenge significantly increased transcript levels of the A(2B) and A(3) receptors, whereas the A(1) receptor message decreased. No change in transcript levels of A(2A) receptors was observed after allergen sensitization and challenge. These findings suggest that A(2B) and A(3) adenosine receptors play an important role in adenosine-induced bronchoconstriction in our allergic mouse model. Finally, whether the airway effects of the receptor agonists/antagonists are direct or indirect needs further investigations. PMID- 12533442 TI - Early surfactant administration protects against lung dysfunction in a mouse model of ARDS. AB - Sepsis can predispose the lung to insults such as mechanical ventilation (MV). It was hypothesized that treating the lung with exogenous surfactant early in the development of sepsis will reduce the lung dysfunction associated with MV 18 h later. Mice underwent sham or cecal ligation and perforation (CLP) surgery. Immediately after surgery, mice were either untreated or given 100 mg/kg of bovine lipid extract surfactant intratracheally. Eighteen hours later, the lungs were removed and analyzed either immediately or following ventilation ex vivo for 2 h by an "injurious" mode of ventilation (20 ml/kg, 0 cm positive end-expiratory pressure). In nonventilated lungs, exogenous surfactant had no impact on compliance or IL-6 concentrations in the lungs. In the ventilated groups, the administered surfactant had a significant protective effect on the lung dysfunction induced by MV, but only in the CLP lungs. We conclude that administration of exogenous surfactant at the time of a systemic insult can protect the lung from the damaging effects of MV 18 h later. PMID- 12533443 TI - Regulation of mucin secretion from human bronchial epithelial cells grown in murine hosted xenografts. AB - Studies of regulated mucin secretion from goblet cells in primary cultures of human bronchial epithelial (HBE) cells have suffered, generally, from poor signal to-noise ratios, with reported secretory responses of <100% (less than onefold) relative to baseline. Using, instead, HBE cells grown as xenografts in the backs of nude mice, we found that UTP (100 micro M) stimulated strong mucin secretory responses from isolated, luminally perfused preparations. The peak response (10 min) for 11 control experiments (37 xenografts) was 3.3 +/- 0.05-fold relative to baseline, and the time-integrated response (60 min) was 23.4 +/- 0.5-fold. Because responses to ATP and UTP were approximately equal, an apical membrane P2Y(2)-receptor (R) is suggested. Additionally, ADP activated mucin release from HBE xenografts, whereas UDP and 2-methlythio-ADP did not, a pattern of response inconsistent with known purinoceptors. Hence, either a novel receptor to ADP is suggested or there is significant conversion of ADP to ATP by ecto-adenylate kinase activity. Adenosine and a nitric oxide donor were without effect. Consistent with P2Y(2)-R coupling to phospholipase C, HBE xenografts responded to ionomycin and PMA; however, they were recalcitrant to forskolin and chlorophenylthio-cAMP, and to 8-bromo-cGMP. Hence, human airway goblet cells, like those of other species, appear to be regulated primarily via phospholipase C pathways, activated particularly by apical membrane P2Y(2)-R agonists. PMID- 12533444 TI - Resident parking only: rhamnolipids maintain fluid channels in biofilms. PMID- 12533445 TI - Microbial conversion of corn stalks to riches. PMID- 12533446 TI - Novel organization and divergent dockerin specificities in the cellulosome system of Ruminococcus flavefaciens. AB - The DNA sequence coding for putative cellulosomal scaffolding protein ScaA from the rumen cellulolytic anaerobe Ruminococcus flavefaciens 17 was completed. The mature protein exhibits a calculated molecular mass of 90,198 Da and comprises three cohesin domains, a C-terminal dockerin, and a unique N-terminal X domain of unknown function. A novel feature of ScaA is the absence of an identifiable cellulose-binding module. Nevertheless, native ScaA was detected among proteins that attach to cellulose and appeared as a glycosylated band migrating at around 130 kDa. The ScaA dockerin was previously shown to interact with the cohesin containing putative surface-anchoring protein ScaB. Here, six of the seven cohesins from ScaB were overexpressed as histidine-tagged products in E. coli; despite their considerable sequence differences, each ScaB cohesin specifically recognized the native 130-kDa ScaA protein. The binding specificities of dockerins found in R. flavefaciens plant cell wall-degrading enzymes were examined next. The dockerin sequences of the enzymes EndA, EndB, XynB, and XynD are all closely related but differ from those of XynE and CesA. A recombinant ScaA cohesin bound selectively to dockerin-containing fragments of EndB, but not to those of XynE or CesA. Furthermore, dockerin-containing EndB and XynB, but not XynE or CesA, constructs bound specifically to native ScaA. XynE- and CesA derived probes did however bind a number of alternative R. flavefaciens bands, including an approximately 110-kDa supernatant protein expressed selectively in cultures grown on xylan. Our findings indicate that in addition to the ScaA dockerin-ScaB cohesin interaction, at least two distinct dockerin-binding specificities are involved in the novel organization of plant cell wall-degrading enzymes in this species and suggest that different scaffoldins and perhaps multiple enzyme complexes may exist in R. flavefaciens. PMID- 12533447 TI - ISCce1 and ISCce2, two novel insertion sequences in Clostridium cellulolyticum. AB - Two new insertion sequences, ISCce1 and ISCce2, were found to be inserted into the cipC gene of spontaneous mutants of Clostridium cellulolyticum. In these insertional mutants, the cipC gene was disrupted either by ISCce1 alone or by both ISCce1 and ISCce2. ISCce1 is 1,292 bp long and has one open reading frame. The open reading frame encodes a putative 348-amino-acid protein with significant levels of identity with putative proteins having unknown functions and with some transposases belonging to the IS481 and IS3 families. Imperfect 23-bp inverted repeats were found near the extremities of ISCce1. ISCce2 is 1,359 bp long, carries one open reading frame, and has imperfect 35-bp inverted repeats at its termini. The open reading frame encodes a putative 398-amino-acid protein. This protein shows significant levels of identity with transposases belonging to the IS256 family. Upon transposition, both ISCce1 and ISCce2 generate 8-bp direct repeats of the target sequence, but no consensus sequences could be identified at either insertion site. ISCce1 is copied at least 20 times in the genome, as assessed by Southern blot analysis. ISCce2 was found to be mostly inserted into ISCce1. In addition, as neither of the elements was detected in seven other Clostridium species, we concluded that they may be specific to the C. cellulolyticum strain used. PMID- 12533448 TI - Characterization of Helicobacter pylori nickel metabolism accessory proteins needed for maturation of both urease and hydrogenase. AB - Previous studies demonstrated that two accessory proteins, HypA and HypB, play a role in nickel-dependent maturation of both hydrogenase and urease in Helicobacter pylori. Here, the two proteins were purified and characterized. HypA bound two Ni(2+) ions per dimer with positive cooperativity (Hill coefficient, approximately 2.0). The dissociation constants K(1) and K(2) for Ni(2+) were 58 and 1.3 microM, respectively. Studies on purified site-directed mutant proteins in each of the five histidine residues within HypA, revealed that only one histidine residue (His2) is vital for nickel binding. Nuclear magnetic resonance analysis showed that this purified mutant version (H2A) was similar in structure to that of the wild-type HypA protein. A chromosomal site-directed mutant of hypA (in the codon for His2) lacked hydrogenase activity and possessed only 2% of the wild-type urease activity. Purified HypB had a GTPase activity of 5 nmol of GTP hydrolyzed per nmol of HypB per min. Site-directed mutagenesis within the lysine residue in the conserved GTP-binding motif of HypB (Lys59) nearly abolished the GTPase activity of the mutant protein (K59A). In native solution, both HypA and HypB exist as homodimers with molecular masses of 25.8 and 52.4 kDa, respectively. However, a 1:1 molar mixture of HypA plus HypB gave rise to a 43.6 kDa species composed of both proteins. A 43-kDa heterodimeric HypA-HypB complex was also detected by cross-linking. The cross-linked adduct was still observed in the presence of 0.5 mM GTP or 1 microM nickel or when the mutant version of HypA (altered in His2) and HypB (altered in Lys59) were tested. Individually, HypA and HypB formed homodimeric cross-linked adducts. An interaction between HypA and the Hp0868 protein (encoded by the gene downstream of hypA) could not be detected via cross-linking, although such an interaction was predicted by yeast two-hybrid studies. In addition, the phenotype of an insertional mutation within the Hp0868 gene indicated that its presence is not critical for either the urease or the hydrogenase activity. PMID- 12533449 TI - ATP-dependent interactions between Escherichia coli Min proteins and the phospholipid membrane in vitro. AB - Proper placement of the division apparatus in Escherichia coli requires pole-to pole oscillation of the MinC division inhibitor. MinC dynamics involves a membrane association-dissociation cycle that is driven by the activities of the MinD ATPase and the MinE topological specificity factor, which themselves undergo coupled oscillatory localization cycles. To understand the biochemical mechanisms underlying Min protein dynamics, we studied the interactions of purified Min proteins with phospholipid vesicles and the role of ATP in these interactions. We show that (i) the ATP-bound form of MinD (MinD.ATP) readily associates with phospholipid vesicles in the presence of Mg(2+), whereas the ADP-bound form (MinD.ADP) does not; (ii) MinD.ATP binds membrane in a self-enhancing fashion; (iii) both MinC and MinE can be recruited to MinD.ATP-decorated vesicles; (iv) MinE stimulates dissociation of MinD.ATP from the membrane in a process requiring hydrolysis of the nucleotide; and (v) MinE stimulates dissociation of MinC from MinD.ATP-membrane complexes, even when ATP hydrolysis is blocked. The results support and extend recent work by Z. Hu et al. (Z. Hu, E. P. Gogol, and J. Lutkenhaus, Proc. Natl. Acad. Sci. USA 99:6761-6766, 2002) and support models of protein oscillation wherein MinE induces Min protein dynamics by stimulating the conversion of the membrane-bound form of MinD (MinD.ATP) to the cytoplasmic form (MinD.ADP). The results also indicate that MinE-stimulated dissociation of MinC from the MinC-MinD.ATP-membrane complex can, and may, occur prior to hydrolysis of the nucleotide. PMID- 12533450 TI - 1-methylguanosine-deficient tRNA of Salmonella enterica serovar Typhimurium affects thiamine metabolism. AB - In Salmonella enterica serovar Typhimurium a mutation in the purF gene encoding the first enzyme in the purine pathway blocks, besides the synthesis of purine, the synthesis of thiamine when glucose is used as the carbon source. On carbon sources other than glucose, a purF mutant does not require thiamine, since the alternative pyrimidine biosynthetic (APB) pathway is activated. This pathway feeds into the purine pathway just after the PurF biosynthetic step and upstream of the intermediate 4-aminoimidazolribotide, which is the common intermediate in purine and thiamine synthesis. The activity of this pathway is also influenced by externally added pantothenate. tRNAs from S. enterica specific for leucine, proline, and arginine contain 1-methylguanosine (m(1)G37) adjacent to and 3' of the anticodon (position 37). The formation of m(1)G37 is catalyzed by the enzyme tRNA(m(1)G37)methyltransferase, which is encoded by the trmD gene. Mutations in this gene, which result in an m(1)G37 deficiency in the tRNA, in a purF mutant mediate PurF-independent thiamine synthesis. This phenotype is specifically dependent on the m(1)G37 deficiency, since several other mutations which also affect translation fidelity and induce slow growth did not cause PurF-independent thiamine synthesis. Some antibiotics that are known to reduce the efficiency of translation also induce PurF-independent thiamine synthesis. We suggest that a slow decoding event at a codon(s) read by a tRNA(s) normally containing m(1)G37 is responsible for the PurF-independent thiamine synthesis and that this event causes a changed flux in the APB pathway. PMID- 12533451 TI - Agrobacterium tumefaciens twin-arginine-dependent translocation is important for virulence, flagellation, and chemotaxis but not type IV secretion. AB - This study characterized the contribution of the twin-arginine translocation (TAT) pathway to growth, motility, and virulence of the phytopathogen Agrobacterium tumefaciens. In contrast to wild-type strain A348, a tatC null mutant failed to export the green fluorescent protein fused to the trimethylamine N-oxide reductase (TorA) signal sequence or to grow on nitrate as a sole electron acceptor during anaerobic growth. The tatC mutant displayed defects in growth rate and cell division but not in cell viability, and it also released abundant levels of several proteins into the culture supernatant when grown in rich medium or in vir induction minimal medium. Nearly all A348 cells were highly motile in both rich and minimal media. By contrast, approximately 0.1% of the tatC mutant cells were motile in rich medium, and <0.01% were motile in vir induction medium. Nonmotile tatC mutant cells lacked detectable flagella, whereas motile tatC mutant cells collected from the edge of a motility halo possessed flagella but not because of reversion to a functional TAT system. Motile tatC cells failed to exhibit chemotaxis toward sugars under aerobic conditions or towards nitrate under anaerobic conditions. The tatC mutant was highly attenuated for virulence, only occasionally (approximately 15% of inoculations) inciting formation of small tumors on plants after a prolonged incubation period of 6 to 8 weeks. However, an enriched subpopulation of motile tatC mutants exhibited enhanced virulence compared to the nonmotile variants. Finally, the tatC mutant transferred T-DNA and protein effectors to plant cells and a mobilizable IncQ plasmid to agrobacterial recipients at wild-type levels. Together, our findings establish that, in addition to its role in secretion of folded cofactor-bound enzymes functioning in alternative respiration, the TAT system of A. tumefaciens is an important virulence determinant. Furthermore, this secretion pathway contributes to flagellar biogenesis and chemotactic responses but not to sensory perception of plant signals or the assembly of a type IV secretion system. PMID- 12533452 TI - Development of a gene knockout system for the halophilic archaeon Haloferax volcanii by use of the pyrE gene. AB - So far, the extremely halophilic archaeon Haloferax volcanii has the best genetic tools among the archaea. However, the lack of an efficient gene knockout system for this organism has hampered further genetic studies. In this paper we describe the development of pyrE-based positive selection and counterselection systems to generate an efficient gene knockout system. The H. volacanii pyrE1 and pyrE2 genes were isolated, and the pyrE2 gene was shown to code for the physiological enzyme orotate phosphoribosyl transferase. A DeltapyrE2 strain was constructed and used to isolate deletion mutants by the following two steps: (i) integration of a nonreplicative plasmid carrying both the pyrE2 wild-type gene, as a selectable marker, and a cloned chromosomal DNA fragment containing a deletion in the desired gene; and (ii) excision of the integrated plasmid after selection with 5-fluoroorotic acid. Application of this gene knockout system is described. PMID- 12533453 TI - Sizing the holin lesion with an endolysin-beta-galactosidase fusion. AB - Double-stranded DNA phages require two proteins for efficient host lysis: the endolysin, a muralytic enzyme, and the holin, a small membrane protein. In an event that defines the end of the vegetative cycle, the lambda holin S acts suddenly to permeabilize the membrane. This permeabilization enables the R endolysin to attack the cell wall, after which cell lysis occurs within seconds. A C-terminal fusion of the R endolysin with full-length beta-galactosidase (beta Gal) was tested for lytic competence in the context of the late-gene expression system of an induced lambda lysogen. Under these conditions, the hybrid R-beta Gal product, an active tetrameric beta-Gal greater than 480 kDa in mass, was fully functional in lysis mediated by the S holin. Western blot analysis demonstrated that the lytic competence was not due to the proteolytic release of the endolysin domain of the R-beta-Gal fusion protein. The ability of this massive complex to be released by the S holin suggests that S causes a generalized membrane disruption rather than a regular oligomeric membrane pore. Similar results were obtained with an early lysis variant of the S holin and also in parallel experiments with the T4 holin, T, in an identical lambda context. However, premature holin lesions triggered by depolarization of the membrane were nonpermissive for the hybrid endolysin, indicating that these premature lesions constituted less-profound damage to the membrane. Finally, a truncated T holin functional in lysis with the endolysin is completely incompetent for lysis with the hybrid endolysin. A model for the formation of the membrane lesion within homo-oligomeric rafts of holin proteins is discussed. PMID- 12533454 TI - RmpA2, an activator of capsule biosynthesis in Klebsiella pneumoniae CG43, regulates K2 cps gene expression at the transcriptional level. AB - The rmpA2 gene, which encodes an activator for capsular polysaccharide (CPS) synthesis, was isolated from a 200-kb virulence plasmid of Klebsiella pneumoniae CG43. Based on the sequence homology with LuxR at the carboxyl-terminal DNA binding motif, we hypothesized that RmpA2 exerts its effect by activating the expression of cps genes that are responsible for CPS biosynthesis. Two luxAB transcriptional fusions, each containing a putative promoter region of the K. pneumoniae K2 cps genes, were constructed and were found to be activated in the presence of multicopy rmpA2. The activation is likely due to direct binding of RmpA2 to the cps gene promoter through its C-terminal DNA binding motif. Moreover, the loss of colony mucoidy in a K. pneumoniae strain deficient in RcsB, a regulator for cps gene expression, could be recovered by complementing the strain with a multicopy plasmid carrying rmpA2. The CPS production in Lon protease-deficient K. pneumoniae significantly increased, and the effect was accompanied by an increase of RmpA2 stability. The expression of the rmpA2 gene was negatively autoregulated and could be activated when the organism was grown in M9 minimal medium. An IS3 element located upstream of the rmpA2 was required for the full activation of the rmpA2 promoter. In summary, our results suggest that the enhancement of K2 CPS synthesis in K. pneumoniae CG43 by RmpA2 can be attributed to its transcriptional activation of K2 cps genes, and the expression level of rmpA2 is autoregulated and under the control of Lon protease. PMID- 12533455 TI - Role for both DNA and RNA in GTP hydrolysis by the Neisseria gonorrhoeae signal recognition particle receptor. AB - The prokaryotic signal recognition particle (SRP) targeting system is a complex of two proteins, FtsY and Ffh, and a 4.5S RNA that targets a subset of proteins to the cytoplasmic membrane cotranslationally. We previously showed that Neisseria gonorrhoeae PilA is the gonococcal FtsY homolog. In this work, we isolated the other two components of the gonococcal SRP, Ffh and 4.5S RNA, and characterized the interactions among the three SRP components by using gel retardation and nitrocellulose filter-binding assays and enzymatic analyses of the two proteins. In the current model of prokaryotic SRP function, based on studies of the Escherichia coli and mammalian systems, Ffh binds to 4.5S RNA and the Ffh-4.5S RNA complex binds to the signal sequence of nascent peptides and then docks with FtsY at the membrane. GTP is hydrolyzed by both proteins synergistically, and the nascent peptide is transferred to the translocon. We present evidence that the in vitro properties of the gonococcal SRP differ from those of previously described systems. GTP hydrolysis by PilA, but not that by Ffh, was stimulated by 4.5S RNA, suggesting a direct interaction between PilA and 4.5S RNA that has not been reported in other systems. This interaction was confirmed by gel retardation analyses in which PilA and Ffh, both alone and together, bound to 4.5S RNA. An additional novel finding was that P(pilE) DNA, previously shown by us to bind PilA in vitro, also stimulates PilA GTP hydrolysis. On the basis of these data, we hypothesize that DNA may play a role in targeting proteins via the SRP. PMID- 12533456 TI - Quorum sensing in Rhizobium sp. strain NGR234 regulates conjugal transfer (tra) gene expression and influences growth rate. AB - Rhizobium sp. strain NGR234 forms symbiotic, nitrogen-fixing nodules on a wide range of legumes via functions largely encoded by the plasmid pNGR234a. The pNGR234a sequence revealed a region encoding plasmid replication (rep) and conjugal transfer (tra) functions similar to those encoded by the rep and tra genes from the tumor-inducing (Ti) plasmids of Agrobacterium tumefaciens, including homologues of the Ti plasmid quorum-sensing regulators TraI, TraR, and TraM. In A. tumefaciens, TraI, a LuxI-type protein, catalyzes synthesis of the acylated homoserine lactone (acyl-HSL) N-3-oxo-octanoyl-L-homoserine lactone (3 oxo-C8-HSL). TraR binds 3-oxo-C8-HSL and activates expression of Ti plasmid tra and rep genes, increasing conjugation and copy number at high population densities. TraM prevents this activation under noninducing conditions. Although the pNGR234a TraR, TraI, and TraM appear to function similarly to their A. tumefaciens counterparts, the TraR and TraM orthologues are not cross-functional, and the quorum-sensing systems have differences. NGR234 TraI synthesizes an acyl HSL likely to be 3-oxo-C8-HSL, but traI mutants and a pNGR234a-cured derivative produce low levels of a similar acyl-HSL and another, more hydrophobic signal molecule. TraR activates expression of several pNGR234a tra operons in response to 3-oxo-C8-HSL and is inhibited by TraM. However, one of the pNGR234a tra operons is not activated by TraR, and conjugal efficiency is not affected by TraR and 3-oxo-C8-HSL. The growth rate of NGR234 is significantly decreased by TraR and 3-oxo-C8-HSL through functions encoded elsewhere in the NGR234 genome. PMID- 12533457 TI - Beta-lactamase genes of the penicillin-susceptible Bacillus anthracis Sterne strain. AB - Susceptibility to penicillin and other beta-lactam-containing compounds is a common trait of Bacillus anthracis. Beta-lactam agents, particularly penicillin, have been used worldwide to treat anthrax in humans. Nonetheless, surveys of clinical and soil-derived strains reveal penicillin G resistance in 2 to 16% of isolates tested. Bacterial resistance to beta-lactam agents is often mediated by production of one or more types of beta-lactamases that hydrolyze the beta-lactam ring, inactivating the antimicrobial agent. Here, we report the presence of two beta-lactamase (bla) genes in the penicillin-susceptible Sterne strain of B. anthracis. We identified bla1 by functional cloning with Escherichia coli. bla1 is a 927-nucleotide (nt) gene predicted to encode a protein with 93.8% identity to the type I beta-lactamase gene of Bacillus cereus. A second gene, bla2, was identified by searching the unfinished B. anthracis chromosome sequence database of The Institute for Genome Research for open reading frames (ORFs) predicted to encode beta-lactamases. We found a partial ORF predicted to encode a protein with significant similarity to the carboxy-terminal end of the type II beta-lactamase of B. cereus. DNA adjacent to the 5' end of the partial ORF was cloned using inverse PCR. bla2 is a 768-nt gene predicted to encode a protein with 92% identity to the B. cereus type II enzyme. The bla1 and bla2 genes confer ampicillin resistance to E. coli and Bacillus subtilis when cloned individually in these species. The MICs of various antimicrobial agents for the E. coli clones indicate that the two beta-lactamase genes confer different susceptibility profiles to E. coli; bla1 is a penicillinase, while bla2 appears to be a cephalosporinase. The beta-galactosidase activities of B. cereus group species harboring bla promoter-lacZ transcriptional fusions indicate that bla1 is poorly transcribed in B. anthracis, B. cereus, and B. thuringiensis. The bla2 gene is strongly expressed in B. cereus and B. thuringiensis and weakly expressed in B. anthracis. Taken together, these data indicate that the bla1 and bla2 genes of the B. anthracis Sterne strain encode functional beta-lactamases of different types, but gene expression is usually not sufficient to confer resistance to beta lactam agents. PMID- 12533458 TI - Elucidation of the Vibrio anguillarum genetic response to the potential fish probiont Pseudomonas fluorescens AH2, using RNA-arbitrarily primed PCR. AB - The antagonistic interaction between a potential fish probiont, Pseudomonas fluorescens strain AH2, and its target organism, Vibrio anguillarum, was investigated by studying the genetic response of the target organism when it was exposed to the antagonist. We compared the differential display of arbitrarily PCR-amplified gene transcripts in V. anguillarum serotype O1 when it was exposed to AH2 supernatant with the display of transcripts in nonexposed control cultures. Growth of V. anguillarum was immediately arrested when the organism was exposed to 50% (vol/vol) AH2 supernatant. A total of 10 potentially differentially expressed transcripts were identified. Among these we identified a gene homologous to rpoS that was induced in a dose-dependent manner when V. anguillarum was cultured in media supplemented with sterile filtered supernatant from AH2. rpoS was also induced when growth was arrested with the iron chelator 2,2-dipyridyl. A chromosomal transcript homologous to vibE that participates in vibriobactin synthesis in Vibrio cholerae was also upregulated during AH2 exposure. This transcript could represent a functionally active gene in V. anguillarum involved in biosynthesis of anguibactin or another V. anguillarum siderophore. On the pJM1 plasmid of V. anguillarum serotype O1, a pseudogene designated open reading frame E (ORF E) that contains a frameshift mutation was previously identified. The gene homologous to vibE identified in this study, interestingly, also has significant homology to ORF E on the amino acid level and does not possess the frameshift mutation. Thus, the chromosomally encoded vibE homologue could fulfil the role of the inactive plasmid-encoded ORF E pseudogene. Addition of Fe(3+) to the system eliminated the growth arrest, and the genes homologous to rpoS and vibE were not induced. To our knowledge, this is the first study linking rpoS induction to iron starvation. Taken together, the results of this study suggest that a major part of the antagonistic property exhibited by strain AH2 is caused by the ability of siderophores in the supernatant to efficiently chelate iron, which results in instant iron deprivation of the pathogen V. anguillarum and complete growth arrest. PMID- 12533459 TI - The Escherichia coli BarA-UvrY two-component system is needed for efficient switching between glycolytic and gluconeogenic carbon sources. AB - The Escherichia coli BarA and UvrY proteins were recently demonstrated to constitute a novel two-component system, although its function has remained largely elusive. Here we show that mutations in the sensor kinase gene, barA, or the response regulator gene, uvrY, in uropathogenic E. coli drastically affect survival in long-term competition cultures. Using media with gluconeogenic carbon sources, the mutants have a clear growth advantage when competing with the wild type, but using media with carbon sources feeding into the glycolysis leads to a clear growth advantage for the wild type. Results from competitions with mutants in the carbon storage regulation system, CsrA/B, known to be a master switch between glycolysis and gluconeogenesis, led us to propose that the BarA-UvrY two component system controls the Csr system. Taking these results together, we propose the BarA-UvrY two-component system is crucial for efficient adaptation between different metabolic pathways, an essential function for adaptation to a new environment. PMID- 12533460 TI - CcpA-independent regulation of expression of the Mg2+ -citrate transporter gene citM by arginine metabolism in Bacillus subtilis. AB - Transcriptional regulation of the Mg(2+)-citrate transporter, CitM, the main citrate uptake system of Bacillus subtilis, was studied during growth in rich medium. Citrate in the growth medium was required for induction under all growth conditions. In Luria-Bertani medium containing citrate, citM expression was completely repressed during the exponential growth phase, marginally expressed in the transition phase, and highly expressed in the stationary growth phase. The repression was relieved when the cells were grown in spent Luria-Bertani medium. The addition of a mixture of 18 amino acids restored repression. L-Arginine in the mixture appeared to be solely responsible for the repression, and ornithine appeared to be an equally potent repressor of citM expression. Studies of mutant strains deficient in RocR and SigL, proteins required for the expression of the enzymes of the arginase pathway, confirmed that uptake into the cell and, most likely, conversion of arginine to ornithine were required for repression. Arginine-mediated repression was independent of a functional CcpA, the global regulator protein in carbon catabolite repression (CCR). Nevertheless, CCR mediated repression was the major mechanism controlling the expression during exponential growth, while the newly described, CcpA-independent arginine-mediated repression was specifically apparent during the transition phase of growth. PMID- 12533461 TI - 2,5-dialkylresorcinol biosynthesis in Pseudomonas aurantiaca: novel head-to-head condensation of two fatty acid-derived precursors. AB - 2-Hexyl-5-propylresorcinol is the predominant analog of several dialkylresorcinols produced by Pseudomonas aurantiaca (Pseudomonas fluorescens BL915). We isolated and characterized three biosynthetic genes that encode an acyl carrier protein, a beta-ketoacyl-acyl carrier protein synthase III, and a protein of unknown function, all of which collectively allow heterologous production of 2-hexyl-5-propylresorcinol in Escherichia coli. Two regulatory genes exhibiting similarity to members of the AraC family of transcriptional regulators are also present in the identified gene cluster. Based on the deduced functions of the proteins encoded by the gene cluster and the observed incorporation of labeled carbons from octanoic acid into 2-hexyl-5 propylresorcinol, we propose that dialkylresorcinols are derived from medium chain-length fatty acids by an unusual head-to-head condensation of beta-ketoacyl thioester intermediates. Genomic evidence suggests that there is a similar pathway for the biosynthesis of the flexirubin-type pigments in certain bacteria belonging to the order Cytophagales. PMID- 12533462 TI - Characterization, expression, and mutation of the Lactococcus lactis galPMKTE genes, involved in galactose utilization via the Leloir pathway. AB - A cluster containing five similarly oriented genes involved in the metabolism of galactose via the Leloir pathway in Lactococcus lactis subsp. cremoris MG1363 was cloned and characterized. The order of the genes is galPMKTE, and these genes encode a galactose permease (GalP), an aldose 1-epimerase (GalM), a galactokinase (GalK), a hexose-1-phosphate uridylyltransferase (GalT), and a UDP-glucose 4 epimerase (GalE), respectively. This genetic organization reflects the order of the metabolic conversions during galactose utilization via the Leloir pathway. The functionality of the galP, galK, galT, and galE genes was shown by complementation studies performed with both Escherichia coli and L. lactis mutants. The GalP permease is a new member of the galactoside-pentose-hexuronide family of transporters. The capacity of GalP to transport galactose was demonstrated by using galP disruption mutant strains of L. lactis MG1363. A galK deletion was constructed by replacement recombination, and the mutant strain was not able to ferment galactose. Disruption of the galE gene resulted in a deficiency in cell separation along with the appearance of a long-chain phenotype when cells were grown on glucose as the sole carbon source. Recovery of the wild type phenotype for the galE mutant was obtained either by genetic complementation or by addition of galactose to the growth medium. PMID- 12533463 TI - Postdivisional synthesis of the Sporosarcina ureae DNA translocase SpoIIIE either in the mother cell or in the prespore enables Bacillus subtilis to translocate DNA from the mother cell to the prespore. AB - The differentiation of vegetative cells of Bacillus subtilis into spores involves asymmetric cell division, which precedes complete chromosome partitioning. The DNA translocase SpoIIIE is required to translocate the origin distal 70% of the chromosome from the larger mother cell into the smaller prespore, the two cells that result from the division. We have tested the effect of altering the time and location of SpoIIIE synthesis on spore formation. We have expressed the spoIIIE homologue from Sporosarcina ureae in B. subtilis under the control of different promoters. Expression from either a weak mother cell-specific (sigma(E)) promoter or a weak prespore-specific (sigma(F)) promoter partly complemented the sporulation defect of a spoIIIE36 mutant; however, expression from a strong prespore-specific (sigma(F)) promoter did not. DNA translocation from the mother cell to the prespore was assayed using spoIIQ-lacZ inserted at thrC; transcription of spoIIQ occurs only in the prespore. Translocation of thrC::spoIIQ-lacZ into the prespore occurred efficiently when spoIIIE(Su) was expressed from the weak sigma(E)- or sigma(F)-controlled promoters but not when it was expressed from the strong sigma(F)-controlled promoter. It is speculated that the mechanism directing SpoIIIE insertion into the septum in the correct orientation may accommodate slow postseptational, prespore-specific SpoIIIE synthesis but may be swamped by strong prespore-specific synthesis. PMID- 12533464 TI - Requirements for Cu(A) and Cu-S center assembly of nitrous oxide reductase deduced from complete periplasmic enzyme maturation in the nondenitrifier Pseudomonas putida. AB - Bacterial nitrous oxide (N(2)O) reductase is the terminal oxidoreductase of a respiratory process that generates dinitrogen from N(2)O. To attain its functional state, the enzyme is subjected to a maturation process which involves the protein-driven synthesis of a unique copper-sulfur cluster and metallation of the binuclear Cu(A) site in the periplasm. There are seven putative maturation factors, encoded by nosA, nosD, nosF, nosY, nosL, nosX, and sco. We wanted to determine the indispensable proteins by expressing nos genes from Pseudomonas stutzeri in the nondenitrifying organism Pseudomonas putida. An in silico study of denitrifying bacteria revealed that nosL, nosX (or a homologous gene, apbE), and sco, but not nosA, coexist consistently with the N(2)O reductase structural gene and other maturation genes. Nevertheless, we found that expression of only three maturation factors (periplasmic protein NosD, cytoplasmic NosF ATPase, and the six-helix integral membrane protein NosY) together with nosRZ in trans was sufficient to produce catalytically active holo-N(2)O reductase in the nondenitrifying background. We suggest that these obligatory factors are required for Cu-S center assembly. Using a mutational approach with P. stutzeri, we also studied NosA, the Cu-containing outer membrane protein previously thought to have Cu insertase function, and ScoP, a putative membrane-anchored chaperone for Cu(A) metallation. Both of these were found to be dispensable elements for N(2)O reductase biosynthesis. Our experimental and in silico data were integrated in a model of N(2)O reductase maturation. PMID- 12533465 TI - Plant lectin-like bacteriocin from a rhizosphere-colonizing Pseudomonas isolate. AB - Rhizosphere isolate Pseudomonas sp. strain BW11M1, which belongs to the Pseudomonas putida cluster, secretes a heat- and protease-sensitive bacteriocin which kills P. putida GR12-2R3. The production of this bacteriocin is enhanced by DNA-damaging treatment of producer cells. We isolated a TnMod mutant of strain BW11M1 that had lost the capacity to inhibit the growth of strain GR12-2R3. A wild-type genomic fragment encompassing the transposon insertion site was shown to confer the bacteriocin phenotype when it was introduced into Escherichia coli cells. The bacteriocin structural gene was identified by defining the minimal region required for expression in E. coli. This gene was designated llpA (lectin like putidacin) on the basis of significant homology of its 276-amino-acid product with mannose-binding lectins from monocotyledonous plants. LlpA is composed of two monocot mannose-binding lectin (MMBL) domains. Several uncharacterized bacterial genes encoding diverse proteins containing one or two MMBL domains were identified. A phylogenetic analysis of the MMBL domains present in eukaryotic and prokaryotic proteins assigned the putidacin domains to a new bacterial clade within the MMBL-containing protein family. Heterologous expression of the llpA gene also conveyed bacteriocin production to several Pseudomonas fluorescens strains. In addition, we demonstrated that strain BW11M1 and heterologous hosts secrete LlpA into the growth medium without requiring a cleavable signal sequence. Most likely, the mode of action of this lectin-like bacteriocin is different from the modes of action of previously described Pseudomonas bacteriocins. PMID- 12533466 TI - Heme-responsive transcriptional activation of Bordetella bhu genes. AB - Bordetella pertussis and Bordetella bronchiseptica, gram-negative respiratory pathogens of mammals, possess a heme iron utilization system encoded by the bhuRSTUV genes. Preliminary evidence suggested that expression of the BhuR heme receptor was stimulated by the presence of heme under iron-limiting conditions. The hurIR (heme uptake regulator) genes were previously identified upstream of the bhuRSTUV gene cluster and are predicted to encode homologs of members of the iron starvation subfamily of extracytoplasmic function (ECF) regulators. In this study, B. pertussis and B. bronchiseptica DeltahurI mutants, predicted to lack an ECF sigma factor, were constructed and found to be deficient in the utilization of hemin and hemoglobin. Genetic complementation of DeltahurI strains with plasmid-borne hurI restored wild-type levels of heme utilization. B. bronchiseptica DeltahurI mutant BRM23 was defective in heme-responsive production of the BhuR heme receptor; hurI in trans restored heme-inducible BhuR expression to the mutant and resulted in BhuR overproduction. Transcriptional analyses with bhuR-lacZ fusion plasmids confirmed that bhuR transcription was activated in iron starved cells in response to heme compounds. Heme-responsive bhuR transcription was not observed in mutant BRM23, indicating that hurI is required for positive regulation of bhu gene expression. Furthermore, bhuR was required for heme inducible bhu gene activation, supporting the hypothesis that positive regulation of bhuRSTUV occurs by a surface signaling mechanism involving the heme-iron receptor BhuR. PMID- 12533467 TI - Recombination activity of a distinctive integron-gene cassette system associated with Pseudomonas stutzeri populations in soil. AB - Class 1 integrons have strongly influenced the evolution of multiple antibiotic resistance. Diverse integrons have recently been detected directly in a range of natural environments. In order to characterize the properties of these environmental integrons, we sought to isolate organisms containing integrons from soils, which resulted in the isolation of Pseudomonas stutzeri strain Q. Further isolation efforts targeted at this species resulted in recovery of two other strains (P and BAM). 16S rRNA sequences and chromosome mapping showed that these three strains are very closely related clonal variants in a single genomovar of P. stutzeri. Only strains Q and BAM were found to contain an integron and an associated gene cassette array. The intI and attI components of these strains showed 99 and 90% identity, respectively. The structure of these integrons and their associated gene cassettes was similar to that reported previously for other integron classes. The two integrons contained nonoverlapping sets of cassette associated genes. In contrast, many of the cassette-associated recombination sites in the two integrons were similar and were considered to constitute a distinct subfamily consisting of 59-base element (59-be) recombination sites (the Pseudomonas subfamily). The recombination activity of P. stutzeri integron components was tested in cointegrate assays. IntIPstQ was shown to catalyze site specific recombination between its cognate attI site and 59-be sites from antibiotic resistance gene cassettes. While IntIPstQ did not efficiently mediate recombination between members of the Pseudomonas 59-be subfamily and other 59-be types, the former sites were functional when they were tested with IntI1. We concluded that integrons present in P. stutzeri possess recombination activity and represent a hot spot for genomic diversity in this species. PMID- 12533468 TI - In vivo analysis of HPr reveals a fructose-specific phosphotransferase system that confers high-affinity uptake in Streptomyces coelicolor. AB - HPr, the histidine-containing phosphocarrier protein of the bacterial phosphotransferase system (PTS), serves multiple functions in carbohydrate uptake and carbon source regulation in low-G+C-content gram-positive bacteria and in gram-negative bacteria. To assess the role of HPr in the high-G+C-content gram positive organism Streptomyces coelicolor, the encoding gene, ptsH, was deleted. The ptsH mutant BAP1 was impaired in fructose utilization, while growth on other carbon sources was not affected. Uptake assays revealed that BAP1 could not transport appreciable amounts of fructose, while the wild type showed inducible high-affinity fructose transport with an apparent K(m) of 2 microM. Complementation and reconstitution experiments demonstrated that HPr is indispensable for a fructose-specific PTS activity. Investigation of the putative fruKA gene locus led to identification of the fructose-specific enzyme II permease encoded by the fruA gene. Synthesis of HPr was not specifically enhanced in fructose-grown cells and occurred also in the presence of non-PTS carbon sources. Transcriptional analysis of ptsH revealed two promoters that are carbon source regulated. In contrast to what happens in other bacteria, glucose repression of glycerol kinase was still operative in a ptsH background, which suggests that HPr is not involved in general carbon regulation. However, fructose repression of glycerol kinase was lost in BAP1, indicating that the fructose-PTS is required for transduction of the signal. This study provides the first molecular genetic evidence of a physiological role of the PTS in S. coelicolor. PMID- 12533469 TI - Characterization of a bifunctional archaeal acyl coenzyme A carboxylase. AB - Acyl coenzyme A carboxylase (acyl-CoA carboxylase) was purified from Acidianus brierleyi. The purified enzyme showed a unique subunit structure (three subunits with apparent molecular masses of 62, 59, and 20 kDa) and a molecular mass of approximately 540 kDa, indicating an alpha(4)beta(4)gamma(4) subunit structure. The optimum temperature for the enzyme was 60 to 70 degrees C, and the optimum pH was around 6.4 to 6.9. Interestingly, the purified enzyme also had propionyl-CoA carboxylase activity. The apparent K(m) for acetyl-CoA was 0.17 +/- 0.03 mM, with a V(max) of 43.3 +/- 2.8 U mg(-1), and the K(m) for propionyl-CoA was 0.10 +/- 0.008 mM, with a V(max) of 40.8 +/- 1.0 U mg(-1). This result showed that A. brierleyi acyl-CoA carboxylase is a bifunctional enzyme in the modified 3 hydroxypropionate cycle. Both enzymatic activities were inhibited by malonyl-CoA, methymalonyl-CoA, succinyl-CoA, or CoA but not by palmitoyl-CoA. The gene encoding acyl-CoA carboxylase was cloned and characterized. Homology searches of the deduced amino acid sequences of the 62-, 59-, and 20-kDa subunits indicated the presence of functional domains for carboxyltransferase, biotin carboxylase, and biotin carboxyl carrier protein, respectively. Amino acid sequence alignment of acetyl-CoA carboxylases revealed that archaeal acyl-CoA carboxylases are closer to those of Bacteria than to those of Eucarya. The substrate-binding motifs of the enzymes are highly conserved among the three domains. The ATP binding residues were found in the biotin carboxylase subunit, whereas the conserved biotin-binding site was located on the biotin carboxyl carrier protein. The acyl-CoA-binding site and the carboxybiotin-binding site were found in the carboxyltransferase subunit. PMID- 12533470 TI - A SecE mutation that modulates SecY-SecE translocase assembly, identified as a specific suppressor of SecY defects. AB - The SecY39(Cs) (cold-sensitive) alteration of Arg357 results in a defect of translocation initiation. As a means to dissect the Sec translocation machinery, we isolated mutations that act as suppressors of the secY39 defect. A specific secE mutation, designated secE105, was thus isolated. This mutation proved to be identical with the prlG2 mutation and to suppress a number of cold-sensitive secY mutations. However, other prlG mutations did not effectively suppress the secY defects. Evidence indicates that the Ser105-to-Pro alteration in the C-terminal transmembrane segment of SecE weakens SecY-SecE association. In vitro analyses showed that the SecE(S105P) alteration preferentially stimulates the initial phase of translocation. It is suggested that the S105P alteration affects the SecYEG channel such that it is more prone to open and to accept the translocation initiation domain of a preprotein molecule. PMID- 12533471 TI - Characterization of the stringent response and rel(Bbu) expression in Borrelia burgdorferi. AB - The stringent response is a global bacterial response to nutritional stress mediated by (p)ppGpp. We previously found that both noninfectious Borrelia burgdorferi strain B31 and infectious B. burgdorferi strain N40 produced large amounts of (p)ppGpp during growth in BSK-H medium and suggested that the stringent response was triggered in B. burgdorferi under these conditions. Here we report that (p)ppGpp levels in B. burgdorferi growing in BSK-II or BSK-H medium are not further increased by nutrient limitation or by serine hydroxamate induced inhibition of protein synthesis and that the presence of (p)ppGpp during growth of N40 in BSK-H medium is not associated with decreased 16S rRNA synthesis. Decreased 16S rRNA synthesis was associated with the decreased growth rate of N40 seen during coculture with tick cells, which are growth conditions that were previously shown to decrease (p)ppGpp levels. One-half as much of the mRNA of the gene encoding the Rel protein of B. burgdorferi (rel(Bbu)) was produced by B31 as by N40 during in vitro growth (2 +/- 0.5 and 4 +/- 0.8 fg of rel(Bbu) mRNA/ng of total Borrelia RNA, respectively). Although the amounts of N40 rel(Bbu) mRNA were identical during growth in vitro and in rat peritoneal chambers, they were markedly decreased during growth in nymphal ticks. In contrast to the lack of change in rel(Bbu) mRNA levels, larger amounts of a 78 kDa protein that was cross-reactive with antibodies to Bacillus subtilis Rel(Bsu) were detected in immunoblots of N40 lysates after growth in rat peritoneal chambers than after growth in vitro. Differences in the level of production of (p)ppGpp between B31 and N40 could not be explained by differences in rel(Bbu) promoters since identical transcriptional start sites 309 nucleotides upstream from the B31 and N40 rel(Bbu) ATG start codon and identical sigma(70)-like promoters were identified by primer extension and sequencing analysis. rel(Bbu) complemented an Escherichia coli CF1693 relA spoT double mutant for growth on M9 minimal medium, and the transformed cells produced rel(Bbu) mRNA. These results indicate that rel(Bbu) is functional and that its transcription and translation and production of (p)ppGpp are affected by environmental conditions in strains N40 and B31. They also suggest that in B. burgdorferi, an organism with few rRNA operons that grows slowly, the role of (p)ppGpp may differ from the classic role played by this molecule in E. coli and that (p)ppGpp may not be responsible for growth rate control. PMID- 12533472 TI - Rates and consequences of recombination between rRNA operons. AB - A mutant strain of Escherichia coli was created by inserting a cassette encoding sucrose sensitivity and neomycin resistance (sacB-neo) into the small-subunit rRNA-encoding gene rrs in the rrnB operon. During growth in a complex medium, the cassette was lost from the population, and a complete rrs gene was restored at a rate of 5 x 10(-9) per cell division. Repair of this lesion required flanking regions of DNA that were similar to the six remaining intact rRNA operons and reestablished the full complement of seven rRNA operons. The relative fitness of strains with restored rrnB operons was 1 to 2% higher than that of the mutant strain. The rrnB operon normally contains a spacer region between the 16S and 23S rRNA-encoding genes that is similar in length and tRNA gene content to the spacer in rrnC, -E, and -G. In 2 of the 14 strains in which rrnB was restored, the spacer region had the same length as the spacer region in rrnA, -D, and -H. The requirement for flanking regions of nearly identical DNA and the replication of the spacer region from other rRNA operons during the repair of rrnB suggest that the restoration was accomplished via gene conversion. The rate of gene conversion was 10-fold less than the fixation of point mutations in the same region of the chromosome but was apparently sufficient to homogenize the sequences of rRNA genes in E. coli. These findings are discussed in the context of a conceptual model describing the presence of sequence heterogeneity in coevolving rRNA genes. PMID- 12533473 TI - The absence of FtsH metalloprotease activity causes overexpression of the sigmaW controlled pbpE gene, resulting in filamentous growth of Bacillus subtilis. AB - FtsH is a membrane-bound and energy-dependent metalloprotease in bacteria which is involved in the posttranslational control of the activity of a variety of important transcription factors and in the degradation of uncomplexed integral membrane proteins. For Bacillus subtilis, little is known about the target proteins of FtsH protease. Its gene is not essential, but knockout strains display a pleiotropic phenotype including sensitivity toward salt and heat stress, defects in sporulation and competence, and largely filamentous growth. Comparison of the intracellular proteomes of wild-type and ftsH knockout strains revealed that at least nine proteins accumulated in the absence of ftsH, four of which could be identified. Two of these proteins turned out to be members of the sigma(W) regulon. Accumulation of one of these sigma(W)-controlled proteins, the penicillin-binding protein PBP4*, was analyzed in more detail. We could show that PBP4* is not a proteolytic substrate of FtsH and that its overproduction is due to the enhanced transcription of its gene (pbpE) in ftsH null mutants. The filamentous growth phenotype of DeltaftsH strains was abolished in a DeltaftsH DeltapbpE double knockout. In ftsH wild-type strains with the pbpE gene under regulatable control, pbpE overexpression caused filamentation of the cells. DNA macroarray analysis revealed that most genes of the sigma(W) regulon are transcribed at elevated levels in an ftsH mutant. The influence of FtsH on sigma(W)-controlled genes is discussed. PMID- 12533474 TI - Escherichia coli endoribonucleases involved in cleavage of bacteriophage T4 mRNAs. AB - The dmd mutant of bacteriophage T4 has a defect in growth because of rapid degradation of late-gene mRNAs, presumably caused by mutant-specific cleavages of RNA. Some such cleavages can occur in an allele-specific manner, depending on the translatability of RNA or the presence of a termination codon. Other cleavages are independent of translation. In the present study, by introducing plasmids carrying various soc alleles, we could detect cleavages of soc RNA in uninfected cells identical to those found in dmd mutant-infected cells. We isolated five Escherichia coli mutant strains in which the dmd mutant was able to grow. One of these strains completely suppressed the dmd mutant-specific cleavages of soc RNA. The loci of the E. coli mutations and the effects of mutations in known RNase encoding genes suggested that an RNA cleavage activity causing the dmd mutant specific mRNA degradation is attributable to a novel RNase. In addition, we present evidence that 5'-truncated soc RNA, a stable form in T4-infected cells regardless of the presence of a dmd mutation, is generated by RNase E. PMID- 12533475 TI - Characterization of SrgA, a Salmonella enterica serovar Typhimurium virulence plasmid-encoded paralogue of the disulfide oxidoreductase DsbA, essential for biogenesis of plasmid-encoded fimbriae. AB - Disulfide oxidoreductases are viewed as foldases that help to maintain proteins on productive folding pathways by enhancing the rate of protein folding through the catalytic incorporation of disulfide bonds. SrgA, encoded on the virulence plasmid pStSR100 of Salmonella enterica serovar Typhimurium and located downstream of the plasmid-borne fimbrial operon, is a disulfide oxidoreductase. Sequence analysis indicates that SrgA is similar to DsbA from, for example, Escherichia coli, but not as highly conserved as most of the chromosomally encoded disulfide oxidoreductases from members of the family Enterobacteriaceae. SrgA is localized to the periplasm, and its disulfide oxidoreductase activity is dependent upon the presence of functional DsbB, the protein that is also responsible for reoxidation of the major disulfide oxidoreductase, DsbA. A quantitative analysis of the disulfide oxidoreductase activity of SrgA showed that SrgA was less efficient than DsbA at introducing disulfide bonds into the substrate alkaline phosphatase, suggesting that SrgA is more substrate specific than DsbA. It was also demonstrated that the disulfide oxidoreductase activity of SrgA is necessary for the production of plasmid-encoded fimbriae. The major structural subunit of the plasmid-encoded fimbriae, PefA, contains a disulfide bond that must be oxidized in order for PefA stability to be maintained and for plasmid-encoded fimbriae to be assembled. SrgA efficiently oxidizes the disulfide bond of PefA, while the S. enterica serovar Typhimurium chromosomally encoded disulfide oxidoreductase DsbA does not. pefA and srgA were also specifically expressed at pH 5.1 but not at pH 7.0, suggesting that the regulatory mechanisms involved in pef gene expression are also involved in srgA expression. SrgA therefore appears to be a substrate-specific disulfide oxidoreductase, thus explaining the requirement for an additional catalyst of disulfide bond formation in addition to DsbA of S. enterica serovar Typhimurium. PMID- 12533476 TI - Involvement of a protein tyrosine kinase in production of the polymeric bioemulsifier emulsan from the oil-degrading strain Acinetobacter lwoffii RAG-1. AB - The genes associated with the biosynthesis of the polymeric bioemulsifier emulsan, produced by the oil-degrading Acinetobacter lwoffii RAG-1 are clustered within a 27-kbp region termed the wee cluster. This report demonstrates the involvement of two genes of the wee cluster of RAG-1, wzb and wzc, in emulsan biosynthesis. The two gene products, Wzc and Wzb were overexpressed and purified. Wzc exhibited ATP-dependent autophosphorylating protein tyrosine kinase activity. Wzb was found to be a protein tyrosine phosphatase capable of dephosphorylating the phosphorylated Wzc. Using the synthetic substrate p-nitrophenyl phosphate (PNPP) Wzb exhibited a V(max) of 12 micromol of PNPP min(-1) mg(-1) and a K(m) of 8 mM PNPP at 30 degrees C. The emulsifying activity of mutants lacking either wzb or wzc was 16 and 15% of RAG-1 activity, respectively, suggesting a role for the two enzymes in emulsan production. Phosphorylation of Wzc was found to occur within a cluster of five tyrosine residues at the C terminus. Colonies from a mutant in which these five tyrosine residues were replaced by five phenylalanine residues along with those of a second mutant, which also lacked Wzb, exhibited a highly viscous colony consistency. Emulsan activity of these mutants was 25 and 24% of that of RAG-1, respectively. Neither of these mutants contained cell associated emulsan. However, they did produce an extracellular high-molecular mass galactosamine-containing polysaccharide. A model is proposed in which subunit polymerization, translocation and release of emulsan are all associated and coregulated by tyrosine phosphorylation. PMID- 12533477 TI - The iron-binding protein Dps confers hydrogen peroxide stress resistance to Campylobacter jejuni. AB - We identified and characterized the iron-binding protein Dps from Campylobacter jejuni. Electron microscopic analysis of this protein revealed a spherical structure of 8.5 nm in diameter, with an electron-dense core similar to those of other proteins of the Dps (DNA-binding protein from starved cells) family. Cloning and sequencing of the Dps-encoding gene (dps) revealed that a 450-bp open reading frame (ORF) encoded a protein of 150 amino acids with a calculated molecular mass of 17,332 Da. Amino acid sequence comparison indicated a high similarity between C. jejuni Dps and other Dps family proteins. In C. jejuni Dps, there are iron-binding motifs, as reported in other Dps family proteins. C. jejuni Dps bound up to 40 atoms of iron per monomer, whereas it did not appear to bind DNA. An isogenic dps-deficient mutant was more vulnerable to hydrogen peroxide than its parental strain, as judged by growth inhibition tests. The iron chelator Desferal restored the resistance of the Dps-deficient mutant to hydrogen peroxide, suggesting that this iron-binding protein prevented generation of hydroxyl radicals via the Fenton reaction. Dps was constitutively expressed during both exponential and stationary phase, and no induction was observed when the cells were exposed to H(2)O(2) or grown under iron-supplemented or iron restricted conditions. On the basis of these data, we propose that this iron binding protein in C. jejuni plays an important role in protection against hydrogen peroxide stress by sequestering intracellular free iron and is expressed constitutively to cope with the harmful effect of hydrogen peroxide stress on this microaerophilic organism without delay. PMID- 12533478 TI - Comparative analyses of the complete genome sequences of Pierce's disease and citrus variegated chlorosis strains of Xylella fastidiosa. AB - Xylella fastidiosa is a xylem-dwelling, insect-transmitted, gamma-proteobacterium that causes diseases in many plants, including grapevine, citrus, periwinkle, almond, oleander, and coffee. X. fastidiosa has an unusually broad host range, has an extensive geographical distribution throughout the American continent, and induces diverse disease phenotypes. Previous molecular analyses indicated three distinct groups of X. fastidiosa isolates that were expected to be genetically divergent. Here we report the genome sequence of X. fastidiosa (Temecula strain), isolated from a naturally infected grapevine with Pierce's disease (PD) in a wine grape-growing region of California. Comparative analyses with a previously sequenced X. fastidiosa strain responsible for citrus variegated chlorosis (CVC) revealed that 98% of the PD X. fastidiosa Temecula genes are shared with the CVC X. fastidiosa strain 9a5c genes. Furthermore, the average amino acid identity of the open reading frames in the strains is 95.7%. Genomic differences are limited to phage-associated chromosomal rearrangements and deletions that also account for the strain-specific genes present in each genome. Genomic islands, one in each genome, were identified, and their presence in other X. fastidiosa strains was analyzed. We conclude that these two organisms have identical metabolic functions and are likely to use a common set of genes in plant colonization and pathogenesis, permitting convergence of functional genomic strategies. PMID- 12533479 TI - Rhamnolipid surfactant production affects biofilm architecture in Pseudomonas aeruginosa PAO1. AB - In response to certain environmental signals, bacteria will differentiate from an independent free-living mode of growth and take up an interdependent surface attached existence. These surface-attached microbial communities are known as biofilms. In flowing systems where nutrients are available, biofilms can develop into elaborate three-dimensional structures. The development of biofilm architecture, particularly the spatial arrangement of colonies within the matrix and the open areas surrounding the colonies, is thought to be fundamental to the function of these complex communities. Here we report a new role for rhamnolipid surfactants produced by the opportunistic pathogen Pseudomonas aeruginosa in the maintenance of biofilm architecture. Biofilms produced by mutants deficient in rhamnolipid synthesis do not maintain the noncolonized channels surrounding macrocolonies. We provide evidence that surfactants may be able to maintain open channels by affecting cell-cell interactions and the attachment of bacterial cells to surfaces. The induced synthesis of rhamnolipids during the later stages of biofilm development (when cell density is high) implies an active mechanism whereby the bacteria exploit intercellular interaction and communication to actively maintain these channels. We propose that the maintenance of biofilm architecture represents a previously unrecognized step in the development of these microbial communities. PMID- 12533480 TI - pIIICTX, a predicted CTXphi minor coat protein, can expand the host range of coliphage fd to include Vibrio cholerae. AB - CTXphi is a filamentous bacteriophage that encodes cholera toxin. CTXphi infection of its host bacterium, Vibrio cholerae, requires the toxin-coregulated pilus (TCP) and the products of the V. cholerae tolQRA genes. Here, we have explored the role of OrfU, a predicted CTXphi minor coat protein, in CTXphi infection. Prior to the discovery that it was part of a prophage, orfU was initially described as an open reading frame of unknown function that lacked similarity to known protein sequences. Based on its size and position in the CTXphi genome, we hypothesized that OrfU may function in a manner similar to that of the coliphage fd protein pIII and mediate CTXphi infection as well as playing a role in CTXphi assembly and release. Deletion of orfU from CTXphi dramatically reduced the number of CTXphi virions detected in supernatants from CTXphi-bearing cells. This defect was complemented by expression of orfU in trans, thereby confirming a role for this gene in CTXphi assembly and/or release. To evaluate the requirement for OrfU in CTXphi infection, we introduced fragments of orfU into gIII in an fd derivative to create OrfU-pIII fusions. While fd is ordinarily unable to infect V. cholerae, an fd phage displaying the N-terminal 274 amino acids of OrfU could infect V. cholerae in a TCP- and TolA-dependent fashion. Since our findings indicate that OrfU functions as the CTXphi pIII, we propose to rename OrfU as pIII(CTX). Our data also provide new evidence for a conserved pathway for filamentous phage infection. PMID- 12533481 TI - The VirB4 family of proposed traffic nucleoside triphosphatases: common motifs in plasmid RP4 TrbE are essential for conjugation and phage adsorption. AB - Proteins of the VirB4 family are encoded by conjugative plasmids and by type IV secretion systems, which specify macromolecule export machineries related to conjugation systems. The central feature of VirB4 proteins is a nucleotide binding site. In this study, we asked whether members of the VirB4 protein family have similarities in their primary structures and whether these proteins hydrolyze nucleotides. A multiple-sequence alignment of 19 members of the VirB4 protein family revealed striking overall similarities. We defined four common motifs and one conserved domain. One member of this protein family, TrbE of plasmid RP4, was genetically characterized by site-directed mutagenesis. Most mutations in trbE resulted in complete loss of its activities, which eliminated pilus production, propagation of plasmid-specific phages, and DNA transfer ability in Escherichia coli. Biochemical studies of a soluble derivative of RP4 TrbE and of the full-length homologous protein R388 TrwK revealed that the purified forms of these members of the VirB4 protein family do not hydrolyze ATP or GTP and behave as monomers in solution. PMID- 12533482 TI - Traffic at the tmRNA gene. AB - A partial screen for genetic elements integrated into completely sequenced bacterial genomes shows more significant bias in specificity for the tmRNA gene (ssrA) than for any type of tRNA gene. Horizontal gene transfer, a major avenue of bacterial evolution, was assessed by focusing on elements using this single attachment locus. Diverse elements use ssrA; among enterobacteria alone, at least four different integrase subfamilies have independently evolved specificity for ssrA, and almost every strain analyzed presents a unique set of integrated elements. Even elements using essentially the same integrase can be very diverse, as is a group with an ssrA-specific integrase of the P4 subfamily. This same integrase appears to promote damage routinely at attachment sites, which may be adaptive. Elements in arrays can recombine; one such event mediated by invertible DNA segments within neighboring elements likely explains the monophasic nature of Salmonella enterica serovar Typhi. One of a limited set of conserved sequences occurs at the attachment site of each enterobacterial element, apparently serving as a transcriptional terminator for ssrA. Elements were usually found integrated into tRNA-like sequence at the 3' end of ssrA, at subsites corresponding to those used in tRNA genes; an exception was found at the non-tRNA-like 3' end produced by ssrA gene permutation in cyanobacteria, suggesting that, during the evolution of new site specificity by integrases, tropism toward a conserved 3' end of an RNA gene may be as strong as toward a tRNA-like sequence. The proximity of ssrA and smpB, which act in concert, was also surveyed. PMID- 12533484 TI - Attachment organelle formation represented by localization of cytadherence proteins and formation of the electron-dense core in wild-type and mutant strains of Mycoplasma pneumoniae. AB - Cytadherence proteins of Mycoplasma pneumoniae are localized at the attachment organelle, which is involved in adhesion, gliding motility, and cell division. The localization of these proteins in cytadherence-deficient mutants was examined by immunofluorescence microscopy. In the class I-2 mutant, which has a frameshift mutation in the hmw2 gene, fluorescent foci for HMW1 and HMW3 were found with reduced intensity, and P1 adhesin showed reduced focusing. However, foci for P90, P40, P30, and P65 were not observed in this mutant. In the class IV-22 mutant, which lacks expression of P1, P90, and P40, the other cytadherence proteins (HMW1, HMW3, P30, and P65) were focused. In a mutant lacking HMW1, signals for HMW3, P90, P40, P30, and P65 were not found, and P1 was distributed throughout the cell. These results suggest that HMW1 is essential for the localization of all other cytadherence proteins, while HMW2 is essential for the localization of P90, P40, P30, and P65. The electron-dense core in cytadherence mutants was observed by thin-section electron microscopy, suggesting that its formation depends on HMW1 and HMW2 and that P1 localization occurs independent of the formation of the electron-dense core. Doubly stained preparations visualized by immunofluorescence microscopy showed that the P1 adhesin, P90, and P40 colocalized to a subregion of the attachment organelle in the wild-type strain. HMW1 and HMW3 also colocalized to a different subregion of the attachment organelle, while P30 and P65 localized at more distal ends of cell poles than HMW1 and HMW3. These differences were more pronounced in cytadherence mutants. These results suggest that there are three distinct subcellular protein localization sites in the attachment organelle, which were represented by HMW1 HMW3, P1-P90-P40, and P30-P65. PMID- 12533483 TI - Microarray analysis of global gene expression in mucoid Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa is the dominant pathogen causing chronic respiratory infections in cystic fibrosis (CF). After an initial phase characterized by intermittent infections, a chronic colonization is established in CF upon the conversion of P. aeruginosa to the mucoid, exopolysaccharide alginate overproducing phenotype. The emergence of mucoid P. aeruginosa in CF is associated with respiratory decline and poor prognosis. The switch to mucoidy in most CF isolates is caused by mutations in the mucA gene encoding an anti-sigma factor. The mutations in mucA result in the activation of the alternative sigma factor AlgU, the P. aeruginosa ortholog of Escherichia coli extreme stress sigma factor sigma(E). Because of the global nature of the regulators of mucoidy, we have hypothesized that other genes, in addition to those specific for alginate production, must be induced upon conversion to mucoidy, and their production may contribute to the pathogenesis in CF. Here we applied microarray analysis to identify on the whole-genome scale those genes that are coinduced with the AlgU sigmulon upon conversion to mucoidy. Gene expression profiles of AlgU-dependent conversion to mucoidy revealed coinduction of a specific subset of known virulence determinants (the major protease elastase gene, alkaline metalloproteinase gene aprA, and the protease secretion factor genes aprE and aprF) or toxic factors (cyanide synthase) that may have implications for disease in CF. Analysis of promoter regions of the most highly induced genes (>40-fold, P < or = 10(-4)) revealed a previously unrecognized, putative AlgU promoter upstream of the osmotically inducible gene osmE. This newly identified AlgU dependent promoter of osmE was confirmed by mapping the mRNA 5' end by primer extension. The recognition of genes induced in mucoid P. aeruginosa, other than those associated with alginate biosynthesis, reported here revealed the identity of previously unappreciated factors potentially contributing to the morbidity and mortality caused by mucoid P. aeruginosa in CF. PMID- 12533485 TI - Characterization of the CipA scaffolding protein and in vivo production of a minicellulosome in Clostridium acetobutylicum. AB - The cipA gene encoding the Clostridium acetobutylicum scaffolding protein CipA was cloned and expressed in Escherichia coli. CipA contains an N-terminal signal peptide, a family 3a cellulose-binding domain (CBD), five type I cohesin domains, and six hydrophilic domains. The uniqueness of CipA lies in the enchainment of cohesin domains that are all separated by a hydrophilic domain. Affinity-purified CipA was used in equilibrium-binding experiments to characterize the interaction of CipA with crystalline cellulose. A K(d) of 0.038 micro M and a [C](max) of 0.43 micro mol of CipA bound per g of Avicel were determined. A mini-CipA polypeptide consisting of a CBD3a and two cohesin domains was overexpressed in C. acetobutylicum, yielding the in vivo formation of a minicellulosome. This is to our knowledge the first demonstration of the in vivo assembly of a recombinant minicellulosome. PMID- 12533486 TI - Plasmid DNA supercoiling and gyrase activity in Escherichia coli wild-type and rpoS stationary-phase cells. AB - Stationary-phase cells displayed a distribution of relaxed plasmids and had the ability to recover plasmid supercoiling as soon as nutrients became available. Preexisting gyrase molecules in these cells were responsible for this recovery. Stationary-phase rpoS cells showed a bimodal distribution of plasmids and failed to supercoil plasmids after the addition of nutrients, suggesting that rpoS plays a role in the regulation of plasmid topology during the stationary phase. PMID- 12533487 TI - The NorM efflux pump of Neisseria gonorrhoeae and Neisseria meningitidis recognizes antimicrobial cationic compounds. AB - In Neisseria gonorrhoeae and Neisseria meningitidis, we identified a gene that would encode a protein highly similar to NorM of Vibrio parahaemolyticus (Y. Morita et al., Antimicrob. Agents Chemother. 42:1778-1782, 1998). A nonpolar insertional mutation in either the gonococcal or meningococcal norM gene resulted in increased bacterial sensitivity to compounds harboring a quaternary ammonium on an aromatic ring (e.g., ethidium bromide, acriflavine hydrochloride, 2-N methylellipticinium, and berberine). The presence of point mutations within the 35 region of a putative norM promoter or a likely ribosome binding site resulted in an increased resistance of gonococci and meningococci to the same compounds, as well as to norfloxacin and ciprofloxacin. Structure-activity relationship studies with putative NorM substrates have found that a cationic moiety is essential for NorM recognition. PMID- 12533488 TI - The high-pathogenicity island is absent in human pathogens of Salmonella enterica subspecies I but present in isolates of subspecies III and VI. AB - In this study we tested 74 Salmonella strains of all eight Salmonella groups and were able to demonstrate the presence of two high-pathogenicity island types in strains of Salmonella groups IIIa, IIIb, and VI. Most high-pathogenicity island positive isolates produced yersiniabactin under iron-limited conditions and were positive for the high-molecular-weight proteins HMWP1 and HMWP2. PMID- 12533489 TI - Genetic and biochemical studies of phosphatase activity of PhoR. AB - In Escherichia coli, PhoR is the histidine kinase of the phosphate regulon. It has been postulated that PhoR may function as a phospho-PhoB phosphatase. Experiments with four precise phoR deletion mutants supported this hypothesis and suggested that this activity resides within the histidine phosphorylation domain. This biochemical activity was confirmed by using a separately expressed histidine phosphorylation domain. PMID- 12533490 TI - Expression of primary sigma factor (PSF) and PSF-like sigma factors in the cyanobacterium Synechocystis sp. strain PCC 6803. AB - Large amounts of sigA mRNA, encoding the primary sigma factor (PSF) in Synechocystis sp. strain PCC 6803, accumulated under standard growth conditions, while stress conditions like heat or high salinity led to a rapid decrease in sigA mRNA content. The sigB, sigC, sigD, and sigE genes, encoding PSF-like sigma factors, were under strict physiological control. PMID- 12533491 TI - Chromosomal arm replacement in Streptomyces griseus. AB - UV irradiation of Streptomyces griseus 2247 yielded a new chromosomal deletion mutant, MM9. Restriction and sequencing analysis revealed that homologous recombination between two similar lipoprotein-like open reading frames, which are located 450 and 250 kb from the left and right ends, respectively, caused chromosomal arm replacement. As a result, new 450-kb terminal inverted repeats (TIRs) were formed in place of the original 24-kb TIRs. Frequent homologous recombinations in Streptomyces strains suggest that telomere deletions can usually be repaired by recombinational DNA repair functioning between the intact and deleted TIR sequences on the same chromosome. PMID- 12533492 TI - Awake negative pressure reflex response of the genioglossus in OSA patients and normal subjects. AB - We hypothesized that the response of the genioglossus to negative pressure during wakefulness should be intact in obstructive sleep apnea (OSA) patients despite published evidence showing impairment of the response of palatal muscles (Mortimore IL and Douglas NJ. Am J Respir Crit Care Med 156: 867-873, 1997). Thus the response of the genioglossus to brief nasal negative pressure applications (NPAs) in early inspiration was compared between OSA patients and an age-matched group of normal subjects at two study sites (n = 11 per group in Long Beach, n = 14 per group in Boston). Subjects were studied in the sitting (Long Beach) or supine (Boston) posture, and the genioglossus electromyogram (EMGgg) was measured with an intraoral surface electrode (Long Beach) or intramuscular electrode (Boston). The response of the EMGgg was expressed as the percent change from baseline where the baseline EMGgg was the value at the onset of the NPA. In Long Beach, the EMGgg response was significantly higher in the OSA patients at a lower suction pressure of approximately 10 cmH(2)O (75.2 +/- 8.4 vs. 37.4 +/- 4.0% increase; P < 0.001) but not at a higher suction pressure of approximately 20 cmH(2)O. In Boston, the response in the OSA patients was also greater (107.2 +/- 25.9 vs. 46.3 +/- 8.3%; P < 0.05) at a suction pressure of approximately 13 cmH(2)O. We conclude that the response of the genioglossus to NPA during wakefulness is not impaired in OSA patients compared with normal subjects and is greater at low suction pressures. PMID- 12533493 TI - Local particle deposition patterns may play a key role in the development of lung cancer. AB - The apparent discrepancy between the reported preferential occurrence of bronchial carcinomas in central bronchial airways and current dose estimates for inhaled particles suggests that experimentally observed local accumulations of particles within bronchial airway bifurcations may play a crucial role in lung cancer induction. Here, we computed three-dimensional particle deposition patterns in lobar-segmental airway bifurcations and quantified the resulting inhomogeneous deposition patterns in terms of deposition enhancement factors, which are defined as the ratio of local to average deposition densities. Our results revealed that a small fraction of epithelial cells located at carinal ridges can receive massive doses that may be even a few hundred times higher than the average dose for the whole airway. This lends further credence to the hypothesis that the apparent site selectivity of neoplastic lesions may indeed be caused by the enhanced deposition of toxic particulate matter at bronchial airway bifurcations. PMID- 12533494 TI - Reactive oxygen species in the plasticity of respiratory behavior elicited by chronic intermittent hypoxia. AB - Long-term facilitation (LTF) of breathing elicited by episodic hypoxia (EH) is an extensively studied example of plasticity of respiratory motor behavior. Previous studies employed the paradigm of EH wherein each episode of hypoxia was 5 min. This paradigm is rarely encountered in nature. Brief episodes of hypoxia are encountered frequently with recurrent apneas, wherein hypoxic episodes last a few seconds only. Recent studies suggest that chronic intermittent hypoxia (CIH) represents a form of oxidative stress involving reactive O(2) species. The objectives of the present study were to determine 1) whether acute, repeated, brief EH (15 s) elicit LTF in breathing and 2) whether prior conditioning with CIH modulates acute EH-induced LTF of breathing, and if so whether reactive O(2) species are involved. Experiments were performed on anesthetized, vagotomized, paralyzed, and mechanically ventilated rats, and efferent phrenic nerve activity was monitored as an index of respiratory motor output. In control animals, acute EH (15-s hypoxia; 10 episodes; n = 9) increased minute neural respiration, which persisted during 60 min of the posthypoxic period, suggesting LTF of breathing. EH-induced LTF of respiration was markedly augmented in CIH-conditioned animals (15-s hypoxia, 9 episodes/h, 8 h/day for 10 days; n = 9). By contrast, conditioning with a comparable, cumulative duration of sustained hypoxia (4-h hypoxia; n = 8) did not augment LTF elicited by acute EH. Systemic administration of manganese (III) tetrakis (1-methyl-4-pyridyl) porphyrin pentachloride (5 mg. kg(-1). day(-1) for 10 days), a potent scavenger of O(2)(-)*, prevented CIH induced potentiation of LTF (n = 9). These results demonstrate that 1) acute, brief EH elicits LTF in respiratory motor output; 2) prior conditioning with CIH, but not with comparable, cumulative duration of sustained hypoxia, augments LTF elicited by acute EH; and 3) O(2)(-)* radical scavenger prevents CIH-induced potentiation of LTF of respiration. PMID- 12533495 TI - Effects of VDT tasks with a bright display at night on melatonin, core temperature, heart rate, and sleepiness. AB - The effects of performing video display terminal (VDT) tasks with a bright display (BD) at night on nocturnal salivary melatonin concentration, rectal temperature, heart rate, and sleepiness were examined. Seven healthy male adults performed exciting VDT tasks with a BD and a dark display (DD) and boring VDT tasks with a BD and a DD from 2300 to 0200. The light intensities of the BD and DD were 45 and 15 lx at each subject's eye level, respectively. The exciting VDT task with both BD and DD significantly suppressed the nocturnal decrease in rectal temperature and heart rate and the nocturnal increase in sleepiness. The BD significantly suppressed the nocturnal decrease in rectal temperature during both exciting and boring VDT tasks. The nocturnal salivary melatonin concentration was significantly suppressed by the combination of the exciting task and BD. The results suggest that performing an exciting VDT task with a BD suppresses the nocturnal changes in melatonin concentration and other physiological indicators of human biological clocks. PMID- 12533496 TI - Greater rate of decline in maximal aerobic capacity with age in endurance-trained than in sedentary men. AB - To determine the relation between habitual endurance exercise status and the age associated decline in maximal aerobic capacity [i.e., maximal O(2) consumption (Vo(2 max))] in men, we performed a well-controlled cross-sectional laboratory study on 153 healthy men aged 20-75 yr: 64 sedentary and 89 endurance trained. Vo(2 max) (ml. kg(-1). min(-1)), measured by maximal treadmill exercise, was inversely related to age in the endurance-trained (r = -0.80) and sedentary (r = 0.74) men but was higher in the endurance-trained men at any age. The rate of decline in Vo(2 max) with age (ml. kg(-1). min(-1)) was greater (P < 0.001) in the endurance-trained than in the sedentary men. Whereas the relative rate of decline in Vo(2 max) (percent decrease per decade from baseline levels in young adulthood) was similar in the two groups, the absolute rate of decline in Vo(2 max) was -5.4 and -3.9 ml. kg(-1). min(-). decade(-1) in the endurance-trained and sedentary men, respectively. Vo(2 max) declined linearly across the age range in the sedentary men but was maintained in the endurance-trained men until approximately 50 yr of age. The accelerated decline in Vo(2 max) after 50 yr of age in the endurance-trained men was related to a decline in training volume (r = 0.46, P < 0.0001) and was associated with an increase in 10-km running time (r = 0.84, P < 0.0001). We conclude that the rate of decline in maximal aerobic capacity during middle and older age is greater in endurance-trained men than in their sedentary peers and is associated with a marked decline in O(2) pulse. PMID- 12533497 TI - Effects of muscle metaboreceptor stimulation on cutaneous blood flow from glabrous and nonglabrous skin in mildly heated humans. AB - Given differences in sympathetic innervation to glabrous and nonglabrous skin, we tested the hypothesis that muscle metaboreceptor regulation of cutaneous vascular conductance (CVC) differs between these skin regions. Subjects (n = 21) performed isometric handgrip exercise (IHG; 50% maximal voluntary contraction for 60 s), followed by 2 min of postexercise ischemia. Throughout IHG and postexercise ischemia, CVC was measured from glabrous (palm) and nonglabrous (forearm and chest) regions contralateral to the exercising arm. These procedures were conducted after the subjects had been exposed to an ambient temperature of 35 degrees C and a relative humidity of 50% for 60 min. These thermal conditions were intended to cause slight increases in cutaneous blood flow via sympathetic withdrawal. Esophageal, sublingual, and mean skin temperatures did not change markedly during IHG or postexercise ischemia. During IHG, forearm CVC did not change, chest CVC increased slightly, and palm CVC decreased substantially (from 100 to 34.8 +/- 3.5%; P = 0.001). During muscle metaboreceptor stimulation due to postexercise ischemia, CVC from nonglabrous regions returned to preexercise baselines, whereas CVC at the palm remained below preexercise baseline (68.2 +/- 4.2%; P = 0.001 relative to preexercise baseline). These results indicate that in mildly heated humans muscle metaboreflex stimulation is capable of modulating CVC in glabrous, but not in nonglabrous, skin. PMID- 12533498 TI - Effect of extracellular PO2 on the fall in intracellular PO2 in contracting single myocytes. AB - The purpose of this investigation was to study the effects of altered extracellular Po(2) (Pe(O(2))) on the intracellular Po(2) (Pi(O(2))) response to contractions in single skeletal muscle cells. Single myocytes (n = 12) were dissected from lumbrical muscles of adult female Xenopus laevis and injected with 0.5 mM Pd-meso-tetra(4-carboxyphenyl)porphine for assessment of Pi(O(2)) via phosphorescence quenching. At a Pe(O(2)) of approximately 20 (low), approximately 40 (moderate), and approximately 60 (high) Torr, tetanic contractions were induced at a frequency of 0.67 Hz for approximately 2 min with a 5-min recovery between bouts (blocked order design). The Pi(O(2)) response to contractions was characterized by a time delay followed by a monoexponential decline to steady state (SS) values. The fall in Pi(O(2)) to SS values was significantly greater at each progressively greater Pe(O(2)) (all P < 0.05). The mean response time (time delay + time constant) was significantly faster in the low (35.2 +/- 5.1 s; P < 0.05 vs. high) and moderate (43.3 +/- 6.4 s; P < 0.05 vs. high) compared with high Pe(O(2)) (61.8 +/- 9.4 s) and was correlated positively (r = 0.965) with the net fall in Pi(O(2)). However, the initial rate of change of Pi(O(2)) (calculated as net fall in Pi(O(2))/time constant) was not different (P > 0.05) among Pe(O(2)) trials. These latter data suggest that, over the range of 20-60 Torr, Pe(O(2)) does not play a deterministic role in setting the initial metabolic response to contractions in isolated frog myocytes. Additionally, these results suggest that oxidative phosphorylation in these myoglobin-free myocytes may be compromised by Pe(O(2)) at values nearing 60 Torr. PMID- 12533499 TI - Endothelium-dependent vasodilation in different rat hindlimb skeletal muscles. AB - Few studies have examined potential for endothelium-dependent vasodilation in skeletal muscles of different fiber-type composition. We hypothesized that muscles composed of slow oxidative (SO)- and/or fast oxidative glycolytic (FOG) type fibers have greater potential for endothelium-dependent vasodilation than muscles composed of fast glycolytic (FG)-type fibers. To test this hypothesis, the isolated perfused rat hindlimb preparation was used with a constant-flow, variable-pressure approach. Perfusion pressure was monitored continuously, and muscle-specific flows were determined by using radiolabeled microspheres at four time points: control, at peak effect of acetylcholine (ACh I; 1-2 x 10(-4) M), at peak effect of ACh after infusion of an endothelial inhibitor (ACh II), and at peak effect of sodium nitroprusside (SNP; 4-5 x 10(-4) M). Conductance was calculated by using pressure and flow data. In the SO-type soleus muscle, conductance increased with ACh and SNP, but the increase in conductance with ACh was partially abolished by the endothelial inhibitor N(G)-nitro-l-arginine methyl ester (control, 0.87 +/- 0.19; ACh I, 2.07 +/- 0.29; ACh II, 1.32 +/- 0.15; SNP, 1.76 +/- 0.19 ml. min(-1). 100 g(-1). mmHg(-1); P < 0.05, ACh I and SNP vs. control). In the FOG-type red gastrocnemius muscle, similar findings were obtained (control, 0.64 +/- 0.11; ACh I, 1.36 +/- 0.21; ACh II, 0.73 +/- 0.16; SNP, 1.30 +/- 0.21 ml. min(-1). 100 g(-1). mmHg; P < 0.05, ACh I and SNP vs. control). In the FG-type white gastrocnemius muscle, neither ACh nor SNP increased conductance. Similar findings were obtained when muscles were combined into high- and low-oxidative muscle groups. Indomethacin had no effect on responses to ACh. These data indicate that endothelium-dependent vasodilation is exhibited by high-oxidative, but not low-oxidative, rat skeletal muscle. Furthermore, endothelium-dependent vasodilation in high-oxidative muscle appears to be primarily mediated by nitric oxide. PMID- 12533500 TI - Hemodilutional anemia is associated with increased cerebral neuronal nitric oxide synthase gene expression. AB - Severe hemodilutional anemia may reduce cerebral oxygen delivery, resulting in cerebral tissue hypoxia. Increased nitric oxide synthase (NOS) expression has been identified following cerebral hypoxia and may contribute to the compensatory increase in cerebral blood flow (CBF) observed after hypoxia and anemia. However, changes in cerebral NOS gene expression have not been reported after acute anemia. This study tests the hypothesis that acute hemodilutional anemia causes cerebral tissue hypoxia, triggering changes in cerebral NOS gene expression. Anesthetized rats underwent hemodilution when 30 ml/kg of blood were exchanged with pentastarch, resulting in a final hemoglobin concentration of 51.0 +/- 1.2 g/l (n = 7 rats). Caudate tissue oxygen tension (Pbr(O(2))) decreased transiently from 17.3 +/- 4.1 to 14.4 +/- 4.1 Torr (P < 0.05), before returning to baseline after approximately 20 min. An increase in CBF may have contributed to restoring Pbr(O(2)) by improving cerebral tissue oxygen delivery. An increase in neuronal NOS (nNOS) mRNA was detected by RT-PCR in the cerebral cortex of anemic rats after 3 h (P < 0.05, n = 5). A similar response was observed after exposure to hypoxia. By contrast, no increases in mRNA for endothelial NOS or interleukin 1beta were observed after anemia or hypoxia. Hemodilutional anemia caused an acute reduction in Pbr(O(2)) and an increase in cerebral cortical nNOS mRNA, supporting a role for nNOS in the physiological response to acute anemia. PMID- 12533501 TI - Enhanced sympathoinhibitory response to volume expansion in conscious hindlimb unloaded rats. AB - Prolonged exposure to microgravity or bed rest produces cardiovascular deconditioning, which is characterized by reductions in plasma volume, alterations in autonomic function, and a predisposition toward orthostatic intolerance. Although the precise mechanisms have not been fully elucidated, it is possible that augmented cardiopulmonary reflexes contribute to some of these effects. The purpose of the present study was to test the hypothesis that sympathoinhibitory responses to volume expansion are enhanced in the hindlimb unloaded (HU) rat, a model of cardiovascular deconditioning. Mean arterial blood pressure, heart rate, and renal sympathetic nerve activity (RSNA) responses to isotonic volume expansion (0.9% saline iv, 15% of plasma volume over 5 min) were examined in conscious HU (14 days) and control animals. Volume expansion produced decreases in RSNA in both groups; however, this effect was significantly greater in HU rats (-46 +/- 7 vs. -25 +/- 4% in controls). Animals instrumented for central venous pressure (CVP) did not exhibit differences in CVP responses to volume expansion. These data suggest that enhanced cardiopulmonary reflexes may be involved in the maintenance of reduced plasma volume and contribute to attenuated baroreflex-mediated sympathoexcitation after spaceflight or bed rest. PMID- 12533502 TI - Contribution of mono- and biarticular muscles to extending knee joint moments in runners and cyclists. AB - Motor actions are governed by coordinated activation of mono- and biarticular muscles. This study considered differences in mono- and biarticular knee extensors between runners and cyclists in the context of adaptations to task specific movement requirements. Two hypotheses were tested: 1) the length-at-use hypothesis, which is that muscle adapts to have it operate around optimal length; and 2) the contraction-mode hypothesis, which is that eccentrically active muscles prefer to operate on the ascending limb of the length-force curve. Ten runners and ten cyclists performed maximal, isometric knee extensions on a dynamometer at five knee and four hip joint angles. This approach allowed the separation of the contribution of mono- and biarticular extensors. Three major differences occurred: 1) compared with runners, monoarticular extensors of cyclists reach optimal length at larger muscle length; 2) in runners, optimal length of the biarticular extensor is shifted to larger lengths; and 3) the moment generated by monoarticular extensor was larger in cyclists. Mono- and biarticular extensors respond to different adaptation triggers in runners and cyclists. Monoarticular muscles seem to adapt to the length-at-use, whereas biarticular muscles were found to be sensitive to the contraction-mode hypothesis. PMID- 12533503 TI - Carbohydrate ingestion influences skeletal muscle cytokine mRNA and plasma cytokine levels after a 3-h run. AB - Sixteen experienced marathoners ran on treadmills for 3 h at approximately 70% maximal oxygen consumption (Vo(2 max)) on two occasions while receiving 1 l/h carbohydrate (CHO) or placebo (Pla) beverages. Blood and vastus lateralis muscle biopsy samples were collected before and after exercise. Plasma was analyzed for IL-6, IL-10, IL-1 receptor agonist (IL-1ra), IL-8, cortisol, glucose, and insulin. Muscle was analyzed for glycogen content and relative gene expression of 13 cytokines by using real-time quantitative RT-PCR. Plasma glucose and insulin were higher, and cortisol, IL-6, IL-10, and IL-1ra, but not IL-8, were significantly lower postexercise in CHO vs. Pla. Change in muscle glycogen content did not differ between CHO and Pla (P = 0.246). Muscle cytokine mRNA content was detected preexercise for seven cytokines in this order (highest to lowest): IL-15, TNF-alpha, IL-8, IL-1beta, IL-12p35, IL-6, and IFN-gamma. After subjects ran for 3 h, gene expression above prerun levels was measured for five of these cytokines: IL-1beta, IL-6, and IL-8 (large increases), and IL-10 and TNF alpha (small increases). The increase in mRNA (fold difference from preexercise) was attenuated in CHO (15.9-fold) compared with Pla (35.2-fold) for IL-6 (P = 0.071) and IL-8 (CHO, 7.8-fold; Pla, 23.3-fold; P = 0.063). CHO compared with Pla beverage ingestion attenuates the increase in plasma IL-6, IL-10, and IL-1ra and gene expression for IL-6 and IL-8 in athletes running 3 h at 70% Vo(2 max) despite no differences in muscle glycogen content. PMID- 12533504 TI - Evidence for central command activation of the human insular cortex during exercise. AB - The purpose of this investigation was to determine whether central command activated regions of the insular cortex, independent of muscle metaboreflex activation and blood pressure elevations. Subjects (n = 8) were studied during 1) rest with cuff occlusion, 2) static handgrip exercise (SHG) sufficient to increase mean blood pressure (MBP) by 15 mmHg, and 3) post-SHG exercise cuff occlusion (PECO) to sustain the 15-mmHg blood pressure increase. Data were collected for heart rate, MBP, ratings of perceived exertion and discomfort, and regional cerebral blood flow (rCBF) by using single-photon-emission computed tomography. When time periods were compared when MBP was matched during SHG and PECO, heart rate (7 +/- 3 beats/min; P < 0.05) and ratings of perceived exertion (15 +/- 2 units; P < 0.05) were higher for SHG. During SHG, there were significant increases in rCBF for hand sensorimotor (9 +/- 3%), right inferior posterior insula (7 +/- 3%), left inferior anterior insula (8 +/- 2%), and anterior cingluate regions (6 +/- 2%), not found during PECO. There was significant activation of the inferior (ventral) thalamus and right inferior anterior insular for both SHG and PECO. Although prior studies have shown that regions of the insular cortex can be activated independent of mechanoreflex input, it was not presently assessed. These findings provide evidence that there are rCBF changes within regions of the insular and anterior cingulate cortexes related to central command per se during handgrip exercise, independent of metaboreflex activation and blood pressure elevation. PMID- 12533505 TI - The multifaceted C. elegans major sperm protein: an ephrin signaling antagonist in oocyte maturation. PMID- 12533506 TI - tRNA transfers to the limelight. PMID- 12533507 TI - Translation elongation after assembly of ribosomes on the Cricket paralysis virus internal ribosomal entry site without initiation factors or initiator tRNA. AB - Reconstitution of translation elongation from purified components confirmed that ribosomes that assembled on the Cricket paralysis virus intercistronic internal ribosomal entry site (IRES) without the involvement of initiation factors or initiator tRNA were active in elongation and are, therefore, true initiation complexes. The first elongation cycle occurred without peptide bond formation on 80S ribosomes that did not contain tRNA in the P site. It required elongation factors 1A and 2 and A site-cognate aminoacylated tRNA. Cycloheximide arrested ribosomes on the IRES only after two cycles of elongation, when the first deacylated tRNA reached the E-site after translocation from the A-site. PMID- 12533508 TI - An Eph receptor sperm-sensing control mechanism for oocyte meiotic maturation in Caenorhabditis elegans. AB - During sexual reproduction in most animals, oocytes arrest in meiotic prophase and resume meiosis (meiotic maturation) in response to sperm or somatic cell signals. Despite progress in delineating mitogen-activated protein kinase (MAPK) and CDK/cyclin activation pathways involved in meiotic maturation, it is less clear how these pathways are regulated at the cell surface. The Caenorhabditis elegans major sperm protein (MSP) signals oocytes, which are arrested in meiotic prophase, to resume meiosis and ovulate. We used DNA microarray data and an in situ binding assay to identify the VAB-1 Eph receptor protein-tyrosine kinase as an MSP receptor. We show that VAB-1 and a somatic gonadal sheath cell-dependent pathway, defined by the CEH-18 POU-class homeoprotein, negatively regulate meiotic maturation and MAPK activation. MSP antagonizes these inhibitory signaling circuits, in part by binding VAB-1 on oocytes and sheath cells. Our results define a sperm-sensing control mechanism that inhibits oocyte maturation, MAPK activation, and ovulation when sperm are unavailable for fertilization. MSP domain proteins are found in diverse animal taxa, where they may regulate contact dependent Eph receptor signaling pathways. PMID- 12533509 TI - Senescence, aging, and malignant transformation mediated by p53 in mice lacking the Brca1 full-length isoform. AB - Senescence may function as a two-edged sword that brings unexpected consequences to organisms. Here we provide evidence to support this theory by showing that the absence of the Brca1 full-length isoform causes senescence in mutant embryos and cultured cells as well as aging and tumorigenesis in adult mice. Haploid loss of p53 overcame embryonic senescence but failed to prevent the adult mutant mice from prematurely aging, which included decreased life span, reduced body fat deposition, osteoporosis, skin atrophy, and decreased wound healing. We further demonstrate that mutant cells that escaped senescence had undergone clonal selection for faster proliferation and extensive genetic/molecular alterations, including overexpression of cyclin D1 and cyclin A and loss of p53. These observations provide the first in vivo evidence that links cell senescence to aging due to impaired function of Brca1 at the expense of tumorigenesis. PMID- 12533510 TI - Genetics of dark skin in mice. AB - Chemical mutagenesis in the mouse is a powerful approach for phenotype-driven genetics, but questions remain about the efficiency with which new mutations ascertained by their phenotype can be localized and identified, and that knowledge applied to a specific biological problem. During a global screen for dominant phenotypes in about 30,000 animals, a novel class of pigmentation mutants were identified by dark skin (Dsk). We determined the genetic map location, homozygous phenotype, and histology of 10 new Dsk and 2 new dark coat (Dcc) mutations, and identified mutations in Agouti (Met1Leu, Dcc4), Sox18 (Leu220ter, Dcc1), Keratin 2e (Thr500Pro, Dsk2), and Egfr (Leu863Gln, Dsk5). Cutaneous effects of most Dsk mutations are limited to melanocytes, except for the Keratin 2e and Egfr mutations, in which hyperkeratosis and epidermal thickening precede epidermal melanocytosis by 3-6 wk. The Dsk2 mutation is likely to impair intermediate filament assembly, leading to cytolysis of suprabasal keratinocytes and secondary hyperkeratosis and melanocytosis. The Dsk5 mutation causes increased tyrosine kinase activity and a decrease in steady-state receptor levels in vivo. The Dsk mutations represent genes or map locations not implicated previously in pigmentation, and delineate a developmental pathway in which mutations can be classified on the basis of body region, microscopic site, and timing of pigment accumulation. PMID- 12533511 TI - Proapoptotic BID is required for myeloid homeostasis and tumor suppression. AB - The proper expansion and contraction of hematopoietic cells requires tight regulation of cell death. BID, a "BH3-only" molecule, amplifies death receptor signals connecting the extrinsic to intrinsic pathways by triggering the mitochondrial pathway of apoptosis. Bid-deficient mice, as they age, spontaneously develop a myeloproliferative disorder, which progresses from myeloid hyperplasia to a fatal, clonal malignancy closely resembling chronic myelomonocytic leukemia (CMML). Thus, an apoptotic defect can result in myeloid leukemogenesis. Premalignant Bid-/- myeloid precursor cells are resistant to death receptor-induced apoptosis. Furthermore, a competitive reconstitution assay demonstrates that Bid-deficient long-term repopulating cells give rise to expanded myelomonocytic cells in vivo. Surprisingly, a single BH3-only molecule operating in the extrinsic death receptor pathway proved essential in vivo for physiologic cell death required to maintain myeloid homeostasis. Moreover, progression to CMML indicates that an upstream BH3-only molecule, BID, is required to suppress tumorigenesis. PMID- 12533512 TI - c-MYC apoptotic function is mediated by NRF-1 target genes. AB - A detailed understanding of the signaling pathways by which c-Myc elicits apoptosis has proven elusive. In the current study, we have evaluated whether the activation of the mitochondrial apoptotic signaling pathway is linked to c-Myc induction of a subset of genes involved in mitochondrial biogenesis. Cytochrome c and other nuclear-encoded mitochondrial genes are regulated by the transcription factor nuclear respiratory factor-1 (NRF-1). The consensus binding sequence (T/C)GCGCA(C/T)GCGC(A/G) of NRF-1 includes a noncanonical CA(C/T)GCG Myc:MAX binding site. In this study, we establish a link between the induction of NRF-1 target genes and sensitization to apoptosis on serum depletion. We demonstrate, by using Northern analysis, transactivation assays, and in vitro and in vivo promoter binding assays that cytochrome c is a direct target of c-Myc. Like c Myc, NRF-1 overexpression sensitizes cells to apoptosis on serum depletion. We also demonstrate that selective interference with c-Myc induction of NRF-1 target genes by using a dominant-negative NRF-1 prevented c-Myc-induced apoptosis, without affecting c-Myc-dependent proliferation. These results suggest that c-myc expression leads to mitochondrial dysfunction and apoptosis by deregulating genes involved in mitochondrial function. PMID- 12533513 TI - The Arabidopsis SRR1 gene mediates phyB signaling and is required for normal circadian clock function. AB - Plants possess several photoreceptors to sense the light environment. In Arabidopsis cryptochromes and phytochromes play roles in photomorphogenesis and in the light input pathways that synchronize the circadian clock with the external world. We have identified SRR1 (sensitivity to red light reduced), a gene that plays an important role in phytochrome B (phyB)-mediated light signaling. The recessive srr1 null allele and phyB mutants display a number of similar phenotypes indicating that SRR1 is required for normal phyB signaling. Genetic analysis suggests that SRR1 works both in the phyB pathway but also independently of phyB. srr1 mutants are affected in multiple outputs of the circadian clock in continuous light conditions, including leaf movement and expression of the clock components, CCA1 and TOC1. Clock-regulated gene expression is also impaired during day-night cycles and in constant darkness. The circadian phenotypes of srr1 mutants in all three conditions suggest that SRR1 activity is required for normal oscillator function. The SRR1 gene was identified and shown to code for a protein conserved in numerous eukaryotes including mammals and flies, implicating a conserved role for this protein in both the animal and plant kingdoms. PMID- 12533514 TI - Tbx1 is regulated by tissue-specific forkhead proteins through a common Sonic hedgehog-responsive enhancer. AB - Haploinsufficiency of Tbx1 is likely a major determinant of cardiac and craniofacial birth defects associated with DiGeorge syndrome. Although mice deficient in Tbx1 exhibit pharyngeal and aortic arch defects, the developmental program and mechanisms through which Tbx1 functions are relatively unknown. We identified a single cis-element upstream of Tbx1 that recognized winged helix/forkhead box (Fox)-containing transcription factors and was essential for regulation of Tbx1 transcription in the pharyngeal endoderm and head mesenchyme. The Tbx1 regulatory region was responsive to signaling by Sonic hedgehog (Shh) in vivo. We show that Shh is necessary for aortic arch development, similar to Tbx1, and is also required for expression of Foxa2 and Foxc2 in the pharyngeal endoderm and head mesenchyme, respectively. Foxa2, Foxc1, or Foxc2 could bind and activate transcription through the critical cis-element upstream of Tbx1, and Foxc proteins were required, within their expression domains, for Tbx1 transcription in vivo. We propose that Tbx1 is a direct transcriptional target of Fox proteins and that Fox proteins may serve an intermediary role in Shh regulation of Tbx1. PMID- 12533517 TI - Mitochondrial and nuclear localization of topoisomerase II in the flagellate Bodo saltans (Kinetoplastida), a species with non-catenated kinetoplast DNA. AB - We have studied topoisomerase II (topo II) in the cells of Bodo saltans, a free living bodonid (Kinetoplastida). Phylogenetic analysis based on the sequence of the entire topo II gene, which is a single-copy gene, confirmed that B. saltans is a predecessor of parasitic trypanosomatids. Antibodies generated against either an overexpressed unique C-terminal region of topo II or a synthetic oligopeptide derived from the same region did not cross-react with cell lysates of related trypanosomatids, while they recognized a single specific band in the B. saltans lysate. Immunolocalization experiments using both antibodies showed that topo II is evenly dispersed throughout the kinetoplast. This is in striking difference from the localization of topo II in other flagellates, where it occurs in two antipodal centers flanking the kinetoplast disk. Moreover, the same topo II has a distinct localization in multiple loci at the periphery of the nucleus of B. saltans. With a minicircle probe derived from the conserved region we have shown that all relaxed non-catenated minicircles are confined to the globular kinetoplast DNA bundle. Therefore, in the mitochondrion of this primitive eukaryote topo II does not catenate relaxed DNA circles into a network in vivo, while a decatenating activity is present in partially purified cell lysates. PMID- 12533515 TI - Coactivation of Rac and Rho by Wnt/Frizzled signaling is required for vertebrate gastrulation. AB - Wnt/Frizzled (Fz) signaling controls cell polarity/movements during vertebrate gastrulation via incompletely defined mechanisms. We demonstrated previously that Wnt/Fz activation of Rho, a GTPase and regulator of cytoskeletal architecture, is essential for vertebrate gastrulation. Here we report that in mammalian cells and Xenopus embryos, Wnt/Fz signaling coactivates Rho and Rac, another GTPase and distinct regulator of cytoskeletal architecture. Wnt/Fz activation of Rac is independent of Rho and mediates Wnt/Fz activation of Jun N-terminal kinase (JNK). Dishevelled (Dvl), a cytoplasmic protein downstream of Fz, forms a Wnt-induced complex with Rac independent of the Wnt-induced Dvl-Rho complex. Depletion or inhibition of Rac function perturbs Xenopus gastrulation without affecting Wnt/Fz activation of the Rho or beta-catenin pathway. We propose that parallel activation of Rac and Rho pathways by Wnt/Fz signaling is required for cell polarity and movements during vertebrate gastrulation. PMID- 12533518 TI - tRNA modification by S-adenosylmethionine:tRNA ribosyltransferase-isomerase. Assay development and characterization of the recombinant enzyme. AB - The enzyme S-adenosylmethionine:tRNA ribosyltransferase-isomerase catalyzes the penultimate step in the biosynthesis of the hypermodified tRNA nucleoside queuosine (Q), an unprecedented ribosyl transfer from the cofactor S adenosylmethionine (AdoMet) to a modified-tRNA precursor to generate epoxyqueuosine (oQ). The complexity of the reaction makes it an especially interesting mechanistic problem, and as a foundation for detailed kinetic and mechanistic studies we have carried out the basic characterization of the enzyme. Importantly, to allow for the direct measurement of oQ formation, we have developed protocols for the preparation of homogeneous substrates; specifically, an overexpression system was constructed for tRNA(Tyr) in an E. coli queA deletion mutant to allow for the isolation of large quantities of substrate tRNA, and [U-ribosyl-(14)C]AdoMet was synthesized. The enzyme shows optimal activity at pH 8.7 in buffers containing various oxyanions, including acetate, carbonate, EDTA, and phosphate. Unexpectedly, the enzyme was inhibited by Mg(2+) and Mn(2+) in millimolar concentrations. The steady-state kinetic parameters were determined to be K(m)(AdoMet) = 101.4 microm, K(m)(tRNA) = 1.5 microm, and k(cat) = 2.5 min( 1). A short minihelix RNA was synthesized and modified with the precursor 7 aminomethyl-7-deazaguanine, and this served as an efficient substrate for the enzyme (K(m)(RNA) = 37.7 microm and k(cat) = 14.7 min(-1)), demonstrating that the anticodon stem-loop is sufficient for recognition and catalysis by QueA. PMID- 12533516 TI - Sonic hedgehog-dependent activation of Gli2 is essential for embryonic hair follicle development. AB - Sonic hedgehog (Shh) signaling plays a critical role in hair follicle development and skin cancer, but how it controls these processes remains unclear. Of the three Gli transcription factors involved in transducing Shh signals in vertebrates, we demonstrate here that Gli2 is the key mediator of Shh responses in skin. Similar to Shh(-/-) mice, Gli2(-/-) mutants exhibit an arrest in hair follicle development with reduced cell proliferation and Shh-responsive gene expression, but grossly normal epidermal differentiation. By transgenic rescue experiments, we show that epidermal Gli2 function alone is sufficient to restore hair follicle development in Gli2(-/-) skin. Furthermore, only a constitutively active form of Gli2, but not wild-type Gli2, can activate Shh-responsive gene expression and promote cell proliferation in Shh(-/-) skin. These observations indicate that Shh-dependent Gli2 activator function in the epidermis is essential for hair follicle development. Our data also reveal that Gli2 mediates the mitogenic effects of Shh by transcriptional activation of cyclin D1 and cyclin D2 in the developing hair follicles. Together, our results suggest that Shh dependent Gli2 activation plays a critical role in epithelial homeostasis by promoting proliferation through the transcriptional control of cell cycle regulators. PMID- 12533519 TI - Interaction of HIV reverse transcriptase with structures mimicking recombination intermediates. AB - Interactions between human immunodeficiency virus (HIV) reverse transcriptase (RT) and structures mimicking intermediates proposed to occur during recombination (strand transfer) were investigated. One mechanism proposed for strand transfer is strand exchange in which a homologous RNA (acceptor) "invades" a donor RNA.DNA duplex (replication intermediate) on which DNA synthesis is occurring. The acceptor displaces the donor of the duplex and binds to the DNA. During exchange a transient trimeric structure forms. A model structure was designed with a replication intermediate to which an acceptor RNA was bound. The acceptor was bound to the 5'-end of the DNA over a 54-base region, whereas the donor associated with the DNA 3'-end over a 28-base region. The dimeric constituents of the trimer (acceptor RNA.DNA and donor RNA.DNA) were also constructed. The acceptor RNA.DNA formed a branched structure in this case. Results showed that RT could cleave the RNA portion of all the structures examined. Association with junction substrates was less stable as determined by off-rates. On the trimer, RT cleaved both RNAs but showed a clear preference for cleaving the donor RNA region. This preference was accentuated by HIV nucleocapsid protein (NC). Results suggest that during recombination RT generally associates with the donor-RNA portion of the trimer and the acceptor RNA is protected but not immune from cleavage. The partial protection likely allows the acceptor RNA to more easily complete strand exchange and shield this RNA to provide a means to salvage replication if the DNA were to dissociate from the cleaved donor RNA. PMID- 12533520 TI - The tau95 subunit of yeast TFIIIC influences upstream and downstream functions of TFIIIC.DNA complexes. AB - The yeast transcription factor IIIC (TFIIIC) is organized in two distinct multisubunit domains, tauA and tauB, that are respectively responsible for TFIIIB assembly and stable anchoring of TFIIIC on the B block of tRNA genes. Surprisingly, we found that the removal of tauA by mild proteolysis stabilizes the residual tauB.DNA complexes at high temperatures. Focusing on the well conserved tau95 subunit that belongs to the tauA domain, we found that the tau95 E447K mutation has long distance effects on the stability of TFIIIC.DNA complexes and start site selection. Mutant TFIIIC.DNA complexes presented a shift in their 5' border, generated slow-migrating TFIIIB.DNA complexes upon stripping TFIIIC by heparin or heat treatment, and allowed initiation at downstream sites. In addition, mutant TFIIIC.DNA complexes were highly unstable at high temperatures. Coimmunoprecipitation experiments indicated that tau95 participates in the interconnection of tauA with tauB via its contacts with tau138 and tau91 polypeptides. The results suggest that tau95 serves as a scaffold critical for tauA.DNA spatial configuration and tauB.DNA stability. PMID- 12533521 TI - P63 is expressed in basal and myoepithelial cells of human normal and tumor salivary gland tissues. AB - p63 is essential for epithelial cell survival and may function as an oncogene. We examined by immunohistochemistry p63 expression in human normal and tumor salivary gland tissues. In normal salivary glands, p63 was expressed in the nuclei of myoepithelial and basal duct cells. Among 68 representative salivary gland tumors, 63 displayed p63 reactivity. In all tumor types differentiated towards luminal and myoepithelial lineages (pleomorphic adenomas, basal cell adenomas, adenoid cystic carcinomas, and epithelial-myoepithelial carcinomas), p63 was expressed in myoepithelial cells, whereas luminal cells were always negative. Similarly, in mucoepidermoid carcinomas, basal, intermediate, and squamous cells expressed p63, in contrast to luminal mucous cells. p63 reactivity was also restricted to basal cells in Warthin tumors and oncocytomas. Myoepitheliomas and myoepithelial carcinomas all expressed p63. The only five negative tumors were three of four acinar cell carcinomas and two of three adenocarcinomas. In conclusion, p63 is expressed in the nuclei of normal human salivary gland myoepithelial and basal duct cells. p63 expression is retained in the modified myoepithelial and basal cells of human salivary gland tumors, which suggests a role for p63 in oncogenesis of these complex tumors. PMID- 12533522 TI - Morphological insights into the origin of glomerular endothelial and mesangial cells and their precursors. AB - Glomerular endothelial and mesangial cells may originate from the metanephric mesenchyme. We used the MAb Thy1.1, a mesangial cell marker in the adult rat kidney, and rat endothelial cell markers MAb RECA-1, MAb PECAM-1 (CD31), and MAb Flk-1 as potential markers to characterize the spatial and temporal distribution of mesangial and endothelial cell precursors during nephrogenesis in the rat. At early stages of glomerulogenesis, RECA-1- and Thy1.1-positive cells were detected in the metanephric blastema at 14 days post conception (dpc) embryos and 15 dpc, respectively, with Thy1.1 expression in cells surrounding the ureteric bud. At 17 and 18 dpc, both RECA-1- and Thy1.1-positive cells were found in the cleft of the S-shaped bodies and in the capillary loops of maturing glomeruli. Double staining for BrdU, a marker of proliferation, and for RECA-1 or BrdU and Thy1.1 also localize in the cleft of S-shaped bodies and in glomerular capillary loops at later stages of development. PDGFRbeta co-localizes in cells expressing endothelial or mesangial markers. The data suggest that endothelial and mesangial cell precursors share common markers during the course of glomerulogenesis and that full differentiation of these cells occurs at late stages of glomerular maturation. Thy1.1- and RECA-1-positive cells may be derived from the metanephric blastemal cells at early stages of kidney development. A subpopulation of these Thy1.1- or RECA-1-positive cells may be precursors that can migrate into the cleft of comma and S-shaped bodies and proliferate in situ to form glomerular capillary tufts. PMID- 12533523 TI - Computerized quantification of tissue vascularization using high-resolution slide scanning of whole tumor sections. AB - Assessment of tissue vascularization using immunohistochemical techniques for microvessel detection has been limited by difficulties in generating reproducible quantitative data. The distinction of individual blood vessels and the selection of microscopic fields to be analyzed remain two factors of subjectivity. In this study, we used imaging analysis software and a high-resolution slide scanner for measurement of CD31-immunostained endothelial area (EA) in whole sections of human neuroblastoma xenograft and murine mammary adenocarcinoma tumors. Imaging analysis software provided objective criteria for analysis of sections of different tumors. The use of the criteria on images of entire tumor section acquired with the slide scanner constituted a rapid method to quantify tumor vascularization. Compared with previously described methods, the "hot spot" and the "random fields" methods, EA measurements obtained with our "whole section scanning" method were more reproducible with 8.6% interobserver disagreement for the "whole section scanning" method vs 42.2% and 39.0% interobserver disagreement for the "hot spot" method and the "random fields," respectively. Microvessel density was also measured with the whole section scanning method and provided additional data on the distribution and the size of the blood vessels. Therefore, this method constitutes a time efficient and reproducible method for quantification of tumor vascularization. PMID- 12533524 TI - Enhanced expression of Duffy antigen in the lungs during suppurative pneumonia. AB - Duffy antigen is a chemokine binding protein expressed on the surface of erythrocytes and postcapillary venular endothelial cells. It binds selective CXC and CC chemokines with high affinity. Although Duffy antigen is present in the normal pulmonary vascular bed, it is not known whether its expression is altered by innate inflammatory responses in the lungs. We studied Duffy antigen expression by immunohistochemistry in autopsy lung specimens from 16 cases of suppurative pneumonia, 11 cases of acute lung injury, and seven normal lungs. In lungs with suppurative pneumonia, Duffy antigen was expressed in higher numbers of pre- and postcapillary parenchymal vessels compared to normal specimens or specimens with acute lung injury (p<0.03 and p<0.02, respectively). Lungs with suppurative pneumonia also showed Duffy antigen expression on the alveolar septa, whereas this was a rare finding in normal specimens or in acute lung injury (p<0.02). Furthermore, Duffy antigen labeling of the alveolar septa localized to regions with airspace accumulation of neutrophil-rich exudates. In summary, Duffy antigen expression is increased in the vascular beds and alveolar septa of the lung parenchyma during suppurative pneumonia, suggesting that Duffy antigen may have a functional role in the lung parenchyma during inflammation. PMID- 12533525 TI - Expression of human pendrin in diseased thyroids. AB - We examined pendrin expression in various diseased thyroid tissues by immunohistochemistry (IHC) using antiserum raised against human pendrin and by real-time quantitative RT-PCR. In normal thyroids the antiserum reacted with the apical membrane of follicular cells and its immunoreactivity was faint. In Graves' thyroids, the IHC expression of pendrin appeared in a pattern similar to that of normal thyroids but it was more extensive and stronger, especially in areas showing marked proliferation of follicular cells. The immunoreactivities of pendrin in nodular goiters varied from case to case. In follicular adenomas, pendrin was localized in the follicle-forming parts of the tumor but was negative in trabecular parts. Pendrin was negative in all follicular carcinomas, papillary carcinomas, and in one case of medullary carcinoma. In quantitive mRNA analysis, the relative values of pendrin mRNA were significantly low in papillary carcinoma (p<0.01), whereas the values in other diseased thyroids were not significantly different from those in normal thyroids. These results suggest that pendrin may play a role in thyroid hormone production as the apical porter of chloride/iodide and investigation of pendrin leads to a better understanding of functional aspects of the iodine transportation system in thyroid diseases. PMID- 12533526 TI - Muscle spindles in the deep muscles of the human neck: a morphological and immunocytochemical study. AB - Muscle spindle density is extremely high in the deep muscles of the human neck. However, there is a paucity of information regarding the morphology and immunoreactivity of these muscle spindles. The objective of this study was to investigate the intrafusal fiber content and to assess the myosin heavy chain (MyHC) composition of muscle spindles from human deep neck muscles. In addition to the conventional spindles containing bag(1), bag(2), and chain fibers (b(1)b(2)c spindle), we observed a number of spindles lacking bag(1) (b(2)c spindle) or bag(2) (b(1)c spindle) fibers. Both bag(1) and bag(2) fibers contained slow tonic MyHCs along their entire fiber length and MyHCI, MyHCIIa, embryonic, and alpha-cardiac MyHC isoforms along a variable length of the fibers. Fetal MyHC was present in bag(2) fibers but not in bag(1) fibers. Nuclear chain fibers contained MyHCIIa, embryonic, and fetal isoforms with regional variations. We also compared the present data with our previous results obtained from muscle spindles in human biceps brachii and the first lumbrical muscles. The allotment of numbers of intrafusal fibers and the MyHC composition showed some muscle related differences, suggesting functional specialization in the control of movement among different human muscles. PMID- 12533527 TI - Brush cells of rodent gallbladder and stomach epithelia express neurofilaments. AB - It has been suggested that brush cells (BCs), a distinct type of cell occurring in various epithelia of the respiratory and gastrointestinal tracts, may function as receptor cells. The major characteristics of BCs are a prominent brush border and an unusually highly ordered arrangement of cytoskeletal elements (F-actin, microtubules, and intermediate filaments). In this study we aimed to characterize the nature of the intermediate filaments in BCs by light and electron microscopic immunostaining. Gallbladder and stomach specimens from mice and rats, respectively, were fixed in various solutions, embedded either in paraffin or epoxy resin, and processed for immunodetection. Commercially available, well characterized antibodies against neurofilaments, peripherin, and cytokeratin peptide 18 were used. The polyclonal antiserum cocktail to neurofilaments was applied as a supplement in a double-labeling procedure with anti-actin and anti cytokeratin 18 antibodies. The results demonstrate that the BCs of both organs express two types of intermediate filaments, i.e., neurofilaments and cytokeratin 18 filaments, and that these have a compartmentalized distribution in the cytoplasm. BCs do not express peripherin. The immunodetection of intermediate filaments distinctive for mature neurons in BCs supports their putative receptor function. The co-expression of neurofilaments and cytokeratins is shown for the first time in healthy tissues. PMID- 12533528 TI - An antigen retrieval method using an alkaline solution allows immunoelectron microscopic identification of secretory granules in conventional epoxy-embedded tissue sections. AB - Immunoelectron microscopy using chromogranin A-specific antibodies has been proposed as an efficient technique for identification of secretory granules (SGs) in tumor cells with evidence of apparent neuroendocrine differentiation. Using an antigen retrieval (AR) method, we succeeded in immunolabeling SGs with antibodies in ultrathin sections of routinely processed epoxy-embedded blocks of tissue. Samples of an insulinoma were fixed in 2% glutaraldehyde, postfixed in 1% OsO(4), and embedded in epoxy resin. Ultrathin sections were immunostained with chromogranin A-specific antibodies and gold-conjugated second antibodies. There was no significant labeling in the absence of AR. Neither etching with sodium metaperiodate nor microwave irradiation of ultrathin sections in citrate buffer (pH 6.0) or in EDTA buffer (pH 8.0) was effective in improving the efficiency of immunolabeling. However, ultrathin epoxy-embedded sections that were microwaved in alkaline solution (pH 10) were adequately labeled (5.2 +/- 0.34 particles per SG). Moreover, considerably improved efficiency of immunostaining was achieved by microwaving sections in alkaline solution (pH 10) with subsequent immunostaining at 60C (12.2 +/- 0.51 particles per SG). This method can also be applied to epoxy embedded sections obtained from formalin-fixed, paraffin-embedded blocks of tissue and was even valid for an old epoxy-embedded block of tissue prepared 15 years previously. PMID- 12533529 TI - Quantitative immunohistochemistry by measuring cumulative signal strength accurately measures receptor number. AB - We previously demonstrated that quantitative immunohistochemistry (Q-IHC) performed by measuring the cumulative signal strength of the digital file encoding an image can be used to determine the absolute amount of chromogen present per pixel. We now show that Q-IHC so performed can be used to accurately determine the amount of peptide hormone receptor of interest in archived tissues. To do this we transfected Balb 3T3 fibroblasts with the cDNA encoding the human receptor for gastrin-releasing peptide (GRP), and selected six cell lines stably expressing between 10(2) and 10(6) receptors/cell. These cell lines were fixed in formalin, embedded in paraffin, and treated with antipeptide antibodies against the GRP receptor, followed by DAB chromogen to identify bound antibody. Images were acquired using a 4.9 million pixel digital scanning 24-bit RGB camera, saved in TIFF format, and used for subsequent analysis. Q-IHC was performed after digitally dissecting out the relevant portion of the image for analysis, and processing using a program written in C (available at http://www.uic.edu/com/dom/gastro/Freedownloads.html). Under the conditions defined here, chromogen quantity as determined by Q-IHC tightly correlated with GRP receptor number (r(2)=0.867) in these cell lines. Using the conversion factor identified as a result of these studies, we then determined GRP receptor number on eight randomly selected, archived human colon cancers. Overall GRP receptor expression in colon cancer depended on the degree to which cells within any particular tumor were differentiated, with well-differentiated cells expressing the greatest numbers of receptors (approximately 55,000 +/- 10,000 sites/cell). These studies indicate that Q-IHC can be used to determine receptor quantity in archived tissues and other samples of limited quantity. PMID- 12533530 TI - Localization of a mitochondrial type of NADP-dependent isocitrate dehydrogenase in kidney and heart of rat: an immunocytochemical and biochemical study. AB - We studied the subcellular localization of the mitochondrial type of NADP dependent isocitrate dehydrogenase (ICD1) in rat was immunofluorescence and immunoelectron microscopy and by biochemical methods, including immunoblotting and Nycodenz gradient centrifugation. Antibodies against a 14-amino-acid peptide at the C-terminus of mouse ICD1 was prepared. Immunoblotting analysis of the Triton X-100 extract of heart and kidney showed that the antibodies developed a single band with molecular mass of 45 kD. ICD1 was highly expressed in heart, kidney, and brown fat but only a low level of ICD1 was expressed in other tissues, including liver. Immunofluorescence staining showed that ICD1 was present mainly in mitochondria and, to a much lesser extent, in nuclei. Low but significant levels of activity and antigen of ICD1 were found in nuclei isolated by equilibrium sedimentation. Immunoblotting analysis of subcellular fractions isolated by Nycodenz gradient centrifugation from rat liver revealed that ICD1 signals were exclusively distributed in mitochondrial fractions in which acyl-CoA dehydrogenase was present. Immunofluorescence staining and postembedding electron microscopy demonstrated that ICD1 was confined almost exclusively to mitochondria and nuclei of rat kidney and heart muscle. The results show that ICD1 is expressed in the nuclei in addition to the mitochondria of rat heart and kidney. In the nuclei, the enzyme is associated with heterochromatin. In kidney, ICD1 distributes differentially in the tubule segments. PMID- 12533531 TI - Immunocytochemical localization of secretogranin III in the anterior lobe of male rat pituitary glands. AB - Secretogranin III (SgIII) is one of the acidic secretory proteins, designated as granins, which are specifically expressed in neuronal and endocrine cells. To clarify its precise distribution in the anterior lobe of the rat pituitary gland, we raised a polyclonal antiserum against rat SgIII for immunocytochemical analyses. By immunohistochemistry using semithin sections, positive signals for SgIII were detected intensely in mammotropes and thyrotropes, moderately in gonadotropes and corticotropes, but not in somatotropes. The distribution pattern of SgIII in the pituitary gland was similar to that of chromogranin B (CgB), also of the granin protein family, suggesting that the expressions of these two granins are regulated by common mechanisms. The localization of SgIII in endocrine cells was confirmed by immunoelectron microscopy. In particular, secretory granules of mammotropes and thyrotropes were densely and preferentially co-labeled for SgIII and CgB in their periphery. Moreover, positive signals for SgIII were occasionally found in cells containing both prolactin and TSH in secretory granules. These lines of evidence suggest that SgIII and CgB are closely associated with the secretory granule membrane and that this membrane association might contribute to gathering and anchoring of other soluble constituents to the secretory granule membrane. PMID- 12533532 TI - Lectin histochemistry shows fucosylated glycoconjugates in the primordial germ cells of Xenopus embryos. AB - Previous works have shown that glycoconjugates with terminal fucose (Fuc) are located in the primordial germ cells (PGCs) of some mammals and might play a role in the migration and adhesion processes during development. The aim of this work was to identify the terminal Fuc moieties of Xenopus PGCs by means of three Fuc binding lectins: from asparagus pea (LTA), gorse seed (UEA-I), and orange peel fungus (AAA). The histochemical procedures were also carried out after deglycosylation pretreatments: beta-elimination with NaOH to remove O-linked oligosaccharides; incubation with PNGase F to remove N-linked carbohydrate chains; and incubation with alpha(1,2)- and alpha(1,6)-fucosidase. The PGCs were always negative for LTA and UEA-I, two lectins that have the highest affinity for Fuc alpha(1,2)-linked. However, the PGCs were strongly labeled with AAA, which preferentially binds to Fuc with alpha(1,3) or alpha(1,4) linkages and to Fuc alpha(1,6)-linked to the proximal N-acetylglucosamine. There was fainter labeling with AAA when the sections were preincubated with alpha(1,6)-fucosidase, but the labeling remained strong when the sections were pretreated with alpha(1,2)fucosidase. When the beta-elimination procedure was carried out, the PGC labeling with AAA was slight. If the PNGase F incubation was performed, the PGCs remained moderately positive for AAA. These data suggest that the Xenopus PGCs have Fuc moieties in O- and N-linked oligosaccharides, including Fuc alpha(1,6) linked to the innermost GlcNAc, and that the Fuc was not in alpha(1,2) linkage. PMID- 12533533 TI - Actin-rich spherical extrusion induced in okadaic acid-treated K562 cells by crosslinking of membrane microdomains. AB - Interconnection between surface microdomains and the actin cytoskeleton is vital to various cellular activities. We studied the responses of okadaic acid (OKA) treated K562 leukemia cells to crosslinking of membrane microdomains. Although OKA alone induced clustering of surface-bound F-actin, addition of a biotinylated poly(ethylene glycol) derivative of cholesterol (bPEG-Chol) and subsequent binding of streptavidin (SA) further induced accumulation of the clusters, resulting in the formation of a spherical cell extrusion. This extrusion was also induced by direct crosslinking of a raft marker, CD59, and ganglioside GM1. In addition, we found that knockout of the gene encoding Fyn kinase inhibited formation of the spherical extrusion in murine T-cells. In bPEG-Chol/SA-treated cells, CD59, ganglioside GM1, and clathrin/AP-2 were all accumulated on the surface of the actin-rich extrusion, whereas dynamin and transferrin receptors were unaffected. Intermediate filaments, mitochondria, and other vesicles also accumulated. These results suggest that crosslinking of membrane domains exaggerates the linkage between actin and a defined set of membrane proteins in OKA-treated cells. PMID- 12533534 TI - A photoreactive fluorescent marker for identifying eosinophils and their cytoplasmic granules in tissues. AB - Here we describe a simple histochemical technique that provides an improved approach to identifying eosinophil components in tissues through the formation of photoreactive complexes that produce stable fluorescent emissions. This method worked readily with histological tissue sections 6-60 microm thick, which were fixed in neutral buffered formalin (NBF), and with cell suspensions similarly fixed and unfixed. Deep red (>605 nm) fluorescent emissions were produced by eosinophil-specific granules when exposed to broadband excitation spectra from a 100-W mercury lamp source (510-590 nm), as well as single-wavelength excitations from both an argon laser (488 nm) and a UV-visible laser (514 nm). The fluorophore-granule complex emissions increased in intensity during the first minute of continuous photoexcitation, then remained stable (>10 min). All nonspecific autofluorescence phenomena associated with these tissues were photobleached in the first minute, including areas of background Biebrich scarlet binding where photoreactive complexes were not formed (i.e., collagen), indicating environmental influences on the fluorophore. This technique allows the visualization of eosinophil granules over a greater period of time than is usually permissible with standard fluorescent markers. Therefore, techniques such as confocal microscopy can be utilized to their fullest extent, providing much more detailed information on the location and distribution of the cytoplasmic contents of eosinophils. PMID- 12533535 TI - Differential expression of connexin43 gap junctions in cardiomyocytes isolated from canine thoracic veins. AB - We investigated the phenotypic features of cardiomyocytes, including the gap junctions, in the myocardial sleeve of thoracic veins. Single cardiomyocytes, isolated from the canine pulmonary veins (PV) and superior vena cava (SVC) using digestive enzymes, were examined by immunoconfocal microscopy using antisera against connexin43 (Cx43), Cx40, and other cell markers. The results showed that isolated cardiomyocytes displayed rod shapes of various sizes, ranging from <50 microm to >200 microm in length, and all the cells expressed alpha-actinin and vinculin. Gap junctions made of various amounts of Cx43 and Cx40 were found at the cell borders. These two connexins were extensively co-localized. Comparison between the thoracic veins showed that cells of the SVC contained more Cx43 gap junctions (total Cx43 gap junctions area per cell surface area, 4.0 +/- 0.2% vs 1.5 +/- 0.2%; p<0.01). In addition, for single-nucleus cells, those from the PV were longer (103.7 +/- 3.6 vs 85.0 +/- 3.1 microm; p<0.01) but narrower (14.4 +/- 0.5 vs 16.9 +/- 0.9 microm; p<0.01). In conclusion, canine thoracic veins contain cardiomyocytes with differences in shape and gap junctions, suggesting that the electrical conduction properties may be different between the thoracic veins. PMID- 12533536 TI - Low resolution structures of the retinoid X receptor DNA-binding and ligand binding domains revealed by synchrotron X-ray solution scattering. AB - Nuclear receptors are ligand-inducible transcription factors that share structurally related DNA-binding (DBD) and ligand-binding (LBD) domains. Biochemical and structural studies have revealed the modular nature of DBD and LBD. Nevertheless, the domains function in concert in vivo. While high-resolution crystal structures of nuclear receptor DBDs and LBDs are available, there are no x-ray structural studies of nuclear receptor proteins containing multiple domains. We report the solution structures of the human retinoid X receptor DBD LBD (hRXRalphaDeltaAB) region. We obtained ab initio shapes of hRXRalphaDeltaAB dimer and tetramer to 3.3 and 1.7 nm resolutions, respectively, and established the position and orientation of the DBD and LBD by fitting atomic coordinates of hRXRalpha DBD and LBD. The dimer is U-shaped with DBDs spaced at approximately 2 nm in a head to head orientation forming an angle of about 10 degrees with respect to each other and with an extensive interface area provided by the LBD. The tetramer is a more elongated X-shaped molecule formed by two dimers in head to head arrangement in which the DBDs are extended from the structure and spaced at about 6 nm. The close proximity of DBDs in dimers may facilitate homodimer formation on DNA; however, for the homodimer to bind to a DNA element containing two directly repeated half-sites, one of the DBDs would need to rotate with respect to the other element. By contrast, the separation of DBDs in the tetramers may account for their decreased ability to recognize DNA. PMID- 12533537 TI - Recognition of the intrinsically flexible addiction antidote MazE by a dromedary single domain antibody fragment. Structure, thermodynamics of binding, stability, and influence on interactions with DNA. AB - The Escherichia coli mazEF operon defines a chromosomal addiction module that programs cell death under various stress conditions. It encodes the toxic and long-lived MazF and the labile antidote MazE. The denaturation of MazE is a two state reversible dimer-monomer transition. At lower concentrations the denatured state is significantly populated. This leads to a new aspect of the regulation of MazE concentration, which may decide about the life and death of the cell. Interactions of MazE with a dromedary antibody domain, cAbMaz1 (previously used as a crystallization aid), as well as with promoter DNA were studied using microcalorimetric and spectroscopic techniques. Unique features of cAbMaz1 enable a specific enthalpy-driven recognition of MazE and, thus, a significant stabilization of its dimeric native conformation. The MazE dimer and the MazE dimer-cAbMaz1 complex show very similar binding characteristics with promoter DNA, i.e. three binding sites with apparent affinities in micromolar range and highly exothermic binding accompanied by large negative entropy contributions. A working model for the MazE-DNA assembly is proposed on the basis of the structural and binding data. Both binding and stability studies lead to a picture of MazE solution structure that is significantly more unfolded than the structure observed in a crystal of the MazE-cAbMaz1 complex. PMID- 12533538 TI - Thermal aggregation of ribonuclease A. A contribution to the understanding of the role of 3D domain swapping in protein aggregation. AB - By lyophilizing RNase A from 40% acetic acid solutions, two dimeric aggregates, the "minor" and "major" dimers (named here N-dimer and C-dimer, respectively), form by 3D domain swapping at a ratio of 1:4. Trimeric and tetrameric aggregates are also obtained. The two dimers and the higher oligomers also form without a lyophilization step. By keeping RNase A dissolved at a high concentration (generally 200 mg/ml) in various media at temperatures ranging from 23 to 70 degrees C for times varying from a few minutes to 2 h, various oligomers, in particular the two dimeric conformers, formed in quite different amounts, often inverting their relative quantities depending on the more or less severe unfolding conditions. When unfolding mainly concerned the N terminus of the protein, richer in hydrophilic residues, the N-dimer, formed by 3D domain swapping of the N-terminal alpha-helix of each monomer, prevailed over the C dimer. Under more vigorous denaturing conditions, where also the C terminus of RNase A, richer in hydrophobic amino acids, unfolded, the C-dimer, formed by 3D domain swapping of the C-terminal beta-strand, prevailed over the other, possibly because of the induction to aggregation promoted by the hydrophobic residues present in the C termini of the two monomers. PMID- 12533539 TI - High resolution structure of an alternate form of the ferric ion binding protein from Haemophilus influenzae. AB - The periplasmic iron binding protein of pathogenic Gram-negative bacteria performs an essential role in iron acquisition from transferrin and other iron sources. Structural analysis of this protein from Haemophilus influenzae identified four amino acids that ligand the bound iron: His(9), Glu(57), Tyr(195), and Tyr(196). A phosphate provides an additional ligand, and the presence of a water molecule is required to complete the octahedral geometry for stable iron binding. We report the 1.14-A resolution crystal structure of the iron-loaded form of the H. influenzae periplasmic ferric ion binding protein (FbpA) mutant H9Q. This protein was produced in the periplasm of Escherichia coli and, after purification and conversion to the apo form, was iron-loaded. H9Q is able to bind ferric iron in an open conformation. A surprising finding in the present high resolution structure is the presence of EDTA located at the previously determined anion ternary binding site, where phosphate is located in the wild type holo and apo structures. EDTA contributes four of the six coordinating ligands for iron, with two Tyr residues, 195 and 196, completing the coordination. This is the first example of a metal binding protein with a bound metal.EDTA complex. The results suggest that FbpA may have the ability to bind and transport iron bound to biological chelators, in addition to bare ferric iron. PMID- 12533540 TI - TIA-1 or TIAR is required for DT40 cell viability. AB - TIA-1 and TIAR are a pair of related RNA-binding proteins which have been implicated in apoptosis. We show that chicken DT40 cells with both tia-1 alleles and one tiar allele disrupted (tia-1(-/-)tiar(-/+) cells) are viable. However, their growth and survival in medium containing low serum levels is significantly reduced compared with DT40 cells. The remaining intact tiar allele in tia-1(-/ )tiar(-/+) cells can only be disrupted if TIA-1 expression is first restored to the cells by transfection of a TIA-1 expression vector. We conclude that DT40 cells require either TIA-1 or TIAR for viability. TIA-1 overexpression in tia-1( /-)tiar(-/+) cells leads to a radical drop in TIAR levels, by inducing efficient splicing of two tiar alternative exons carrying in-frame stop codons. In wild type DT40 cells, tiar transcripts including these exons can also be detected. These transcripts increase significantly in abundance in cycloheximide-treated cells, suggesting that splicing of the exons exposes mRNAs to nonsense-mediated mRNA decay. TIA-1 or TIAR depletion leads to a marked drop in splicing of the exons. The human tiar gene contains a corresponding pair of TIA-1-inducible alternative exons, and we show that there is very high sequence conservation between chickens and humans of the exon pair and parts of the flanking introns. The TIA-1/TIAR responsiveness of these alternative tiar exons is likely to be of physiological importance for controlling TIAR levels. PMID- 12533541 TI - A potassium channel-MiRP complex controls neurosensory function in Caenorhabditis elegans. AB - MinK-related peptides (MiRPs) are single transmembrane proteins that associate with mammalian voltage-gated K(+) subunits. Here we report the cloning and functional characterization of a MiRP beta-subunit, MPS-1, and of a voltage-gated pore-forming potassium subunit, KVS-1, from the nematode Caenorhabditis elegans. mps-1 is expressed in chemosensory and mechanosensory neurons and co-localizes with kvs-1 in a subset of these. Inactivation of either mps-1 or kvs-1 by RNA interference (RNAi) causes partially overlapping neuronal defects and results in broad-spectrum neuronal dysfunction, including defective chemotaxis, disrupted mechanotransduction, and impaired locomotion. Inactivation of one subunit by RNAi dramatically suppresses the expression of the partner subunit only in cells where the two proteins co-localize. Co-expression of MPS-1 and KVS-1 in mammalian cells gives rise to a potassium current distinct from the KVS-1 current. Taken together these data indicate that potassium currents constitute a basic determinant for C. elegans neuronal function and unravel a unifying principle of evolutionary significance: that potassium channels in various organisms use MiRPs to generate uniqueness of function with rich variation in the details. PMID- 12533542 TI - Structural impact of the leukemia drug 1-beta-D-arabinofuranosylcytosine (Ara-C) on the covalent human topoisomerase I-DNA complex. AB - 1-beta-d-Arabinofuranosylcytosine (Ara-C) is a potent antineoplastic drug used in the treatment of acute leukemia. Previous biochemical studies indicated the incorporation of Ara-C into DNA reduced the catalytic activity of human topoisomerase I by decreasing the rate of single DNA strand religation by the enzyme by 2-3-fold. We present the 3.1 A crystal structure of human topoisomerase I in covalent complex with an oligonucleotide containing Ara-C at the +1 position of the non-scissile DNA strand. The structure reveals that a hydrogen bond formed between the 2'-hydroxyl of Ara-C and the O4' of the adjacent -1 base 5' to the damage site stabilizes a C3'-endo pucker in the Ara-C arabinose ring. The structural distortions at the site of damage are translated across the DNA double helix to the active site of human topoisomerase I. The free sulfhydryl at the 5' end of the nicked DNA strand in this trapped covalent complex is shifted out of alignment with the 3'-phosphotyrosine linkage at the catalytic tyrosine 723 residue, producing a geometry not optimal for religation. The subtle structural changes caused by the presence of Ara-C in the DNA duplex may contribute to the cytotoxicity of this leukemia drug by prolonging the lifetime of the covalent human topoisomerase I-DNA complex. PMID- 12533543 TI - Fluorescence resonance energy transfer analysis of protein translocase. SecYE from Thermus thermophilus HB8 forms a constitutive oligomer in membranes. AB - SecY and SecE are the two principal translocase subunits that create a channel like pathway for the transit of preprotein across the bacterial cytoplasmic membrane. Here we report the cloning, expression, and purification of the SecYE complex (TSecYE) from a thermophilic bacterium, Thermus thermophilus HB8. Purified TSecYE can be reconstituted into proteoliposomes that function in T. thermophilus SecA (TSecA) dependent preprotein translocation. After the mixing of TSecYE derivatives labeled with either a donor or an acceptor fluorophore during reconstitution, fluorescence resonance energy transfer experiments demonstrated that 2 or more units of TSecYE in the lipid bilayer associate to form a largely non-exchangeable oligomeric structure. PMID- 12533544 TI - Functional regulation of semaphorin receptors by proprotein convertases. AB - PLEXIN genes encode receptors for secreted and membrane-bound semaphorins. It was proposed that the extracellular domain of plexins acts as an inhibitory moiety, preventing receptor activation. Here we show that plexin-B1 and plexin-B2 undergo proteolytic processing in their extracellular portion, thereby converting single chain precursors into non-disulfide-linked, heterodimeric receptors. We demonstrate that plexin processing is mediated by subtilisin-like proprotein convertases, by inhibition with alpha1-antitrypsin Portland, and by mutagenesis of the substrate-cleavage sites. We provide evidence indicating that proprotein convertases cleave plexins in a post-Golgi compartment and, likely, at the cell surface. In addition, we find that both cell surface targeting and proteolytic processing of plexin-B1 depend on protein-protein interaction motifs in the cytoplasmic domain of the receptor. We then show that proteolytic conversion of plexin-B1 into a heterodimeric receptor greatly increases the binding and the functional response to its specific ligand semaphorin 4D/CD100. Thus, we conclude that cleavage by proprotein convertases is a novel regulatory step for semaphorin receptors localized at the cell surface. PMID- 12533545 TI - Sequential autolytic processing activates the zymogen of Arg-gingipain. AB - Most proteases are synthesized as inactive precursors to protect the synthetic machinery of the cell and allow timing of activation. The mechanisms used to render latency are varied but tend to be conserved within protease families. Proteases belonging to the caspase family have a unique mechanism mediated by transitions of two surface loops, and on the basis of conservation of mechanism one would expect this to be preserved by caspase relatives. We have been able to express the full-length precursor of the Arg-specific caspase relative from the bacterium Porphyromonas gingivalis, Arg-gingipain-B, and we show that it contains N- and C-terminal extensions that render a low amount of latency, meaning that the zymogen is substantially active. Three sequential autolytic processing steps at the N and C terminus are required for full activity, and the N-propeptide may serve as an intramolecular chaperone rather than an inhibitory peptide. Each step in activation requires the previous step, and an affinity probe reveals that incremental activity enhancements are achieved in a stepwise manner. PMID- 12533546 TI - Cholesterol is superior to 7-ketocholesterol or 7 alpha-hydroxycholesterol as an allosteric activator for acyl-coenzyme A:cholesterol acyltransferase 1. AB - We compared the abilities of cholesterol versus various oxysterols as substrate and/or as activator for the enzyme acyl-coenzyme A:cholesterol acyltransferase (ACAT), by monitoring the activity of purified human ACAT1 in response to sterols solubilized in mixed micelles or in reconstituted vesicles. The results showed that 5 alpha,6 alpha-epoxycholesterol and 7 alpha-hydroxycholesterol are comparable with cholesterol as the favored substrates, whereas 7-ketocholesterol, 7 beta-hydroxycholesterol, 5 beta,6 beta-epoxycholesterol, and 24(S),25 epoxycholesterol are very poor substrates for the enzyme. We then tested the ability of 7-ketocholesterol as an activator when cholesterol was measured as the substrate, and vice versa. When cholesterol was measured as the substrate, the addition of 7-ketocholesterol could not activate the enzyme. In contrast, when 7 ketocholesterol was measured as the substrate, the addition of cholesterol significantly activated the enzyme and changed the shape of the substrate saturation curve from sigmoidal to essentially hyperbolic. Additional results show that, as an activator, cholesterol is much better than all the oxysterols tested. These results suggest that ACAT1 contains two types of sterol binding sites; the structural requirement for the ACAT activator site is more stringent than it is for the ACAT substrate site. Upon activation by cholesterol, ACAT1 becomes promiscuous toward various sterols as its substrate. PMID- 12533547 TI - Oligomerization of the murine fatty acid transport protein 1. AB - The 63-kDa murine fatty acid transport protein 1 (FATP1) was cloned on the basis of its ability to augment fatty acid import when overexpressed in mammalian cells. The membrane topology of this integral plasma membrane protein does not resemble that of polytopic membrane transporters for other substrates. Western blot analysis of 3T3-L1 adipocytes that natively express FATP1 demonstrate a prominent 130-kDa species as well as the expected 63-kDa FATP1, suggesting that this protein may participate in a cell surface transport protein complex. To test whether FATP1 is capable of oligomerization, we expressed functional FATP1 molecules with different amino- or carboxyl-terminal epitope tags in fibroblasts. These epitope-tagged proteins also form apparent higher molecular weight species. We show that, when expressed in the same cells, differentially tagged FATP1 proteins co-immunoprecipitate. The region between amino acid residues 191 and 475 is sufficient for association of differentially tagged truncated FATP1 constructs. When wild type FATP1 and the non-functional s250a FATP1 mutant are co expressed in COS7 cells, mutant FATP1 has dominant inhibitory function in fatty acid uptake assays. Taken together, these results are consistent with a model in which FATP1 homodimeric complexes play an important role in cellular fatty acid import. PMID- 12533548 TI - Variability of neutral-position casting of the foot. AB - Neutral-position casting of the foot is used for the manufacture of functional foot orthoses, and an accurate cast is widely assumed to be a prerequisite for a good orthotic device. The primary aim of this study was to determine the variability of casting between inexperienced and experienced clinicians and the variability of one experienced clinician taking multiple casts. Ten inexperienced and ten experienced clinicians took a cast of the right foot of a single subject, and a single experienced clinician took ten casts of the same foot. The frontal plane forefoot-to-rearfoot relationship of each cast was determined, and no difference was found in the mean and variances among the three groups. The range of the forefoot-to-rearfoot relationship across all groups was from 10.0 degrees everted to 6.5 degrees inverted, indicating that there is a wide range in neutral position casting of the foot. As outcome studies have reported the successful outcomes of functional foot orthoses, this wide variability may not necessarily be a problem. PMID- 12533549 TI - Fracture of the second metatarsal base. An overlooked cause of chronic midfoot pain. AB - Fracture of the second metatarsal is a cause of chronic midfoot pain that has not been thoroughly examined in the literature. A retrospective review of medical charts and imaging studies was undertaken to investigate this phenomenon. The clinical presentation, differential diagnosis, results of imaging studies, and treatment modalities are described for eight patients with midfoot pain who were treated for a mean of 3.7 months (range, 0 to 12 months) before imaging studies showed a nondisplaced transverse fracture of the second metatarsal base. Initial radiographs indicated fracture in only one patient. Two patients later had surgical bone grafting, two patients had asymptomatic nonunion, and three patients eventually healed. At the last follow-up examination, one patient was continuing treatment with immobilization and electrical bone stimulation. For the seven patients in whom symptoms resolved, the mean recovery period was 14 months (range, 5 to 23 months). Further research is needed to improve treatment and outcomes for this condition. PMID- 12533550 TI - Knowledge of aging and attitudes toward older people. A survey of Australian podiatric medical students. AB - Podiatric medical students in Australia were surveyed to evaluate their reasons for entering podiatric medicine, knowledge of aging, attitudes toward older people, perceptions of treatment efficacy, and desire to specialize in geriatrics. Few students plan to specialize in geriatrics upon graduation (4%), with most preferring general practice (25%) or sports medicine (21%). However, knowledge of aging was good, and students had favorable attitudes toward older people and considered treatment of older people to be effective. Few age- or gender-related effects were observed. It is concluded that students' lack of desire to specialize in geriatrics may be due not to unfavorable perceptions of older people but rather to the low profile and limited development of geriatrics as a specialty area within the podiatric medical profession. PMID- 12533551 TI - External thermoregulation of wounds associated with lower-extremity osteomyelitis. A pilot study. AB - External thermoregulation using noncontact normothermic wound therapy accelerates wound closure by second intention in areas of existing osteomyelitis before surgical excision compared with standard wound care. This pilot study consisted of two arms. The control arm received standard wound care, which resulted in complete ulcer healing at an average of 127 days. The treatment arm received noncontact normothermic wound therapy, which resulted in complete ulcer healing at an average of 59 days, or 54% faster than in the control arm. This new treatment allows the physician to decrease the rate of limb loss and recurrent osteomyelitis by decreasing the morbidity of bone reinfection through the wound bed. There have been no published studies or case presentations addressing thermoregulation in the management of wounds associated with osteomyelitis. Although noncontact normothermic wound therapy is not a direct treatment for osteomyelitis, this new treatment option results in significantly accelerated healing of wounds associated with osteomyelitis. PMID- 12533552 TI - Use of a needleless injection system for digital ring block anesthesia. AB - Digital ring block anesthesia, which is frequently used before surgery for ingrown toenails, is often extremely uncomfortable for patients and can be the most distressing aspect of the procedure. The authors used a novel needleless injection device to induce digital anesthesia before surgery and compared it in terms of patient discomfort and preference with use of a standard needle and syringe for injection in individuals undergoing simultaneous bilateral nail procedures. Use of the needleless device significantly reduced the pain associated with this procedure and was preferred over use of a standard needle and syringe by all individuals. Other potential advantages of a needleless injection system are discussed. PMID- 12533553 TI - Kinetics of high-heeled gait. AB - A within-subject comparative study of walking while wearing low-heeled sports shoes versus high-heeled dress shoes was performed to identify and describe changes in lower-extremity joint kinetics associated with wearing high-heeled shoes during level overground walking. A volunteer sample of 15 unimpaired female subjects recruited from the local community underwent quantitative measurement of sagittal and frontal plane lower-extremity joint function, including angular motion, muscular moment, power, and work. When walking in high-heeled shoes, a significant reduction in ankle plantar flexor muscle moment, power, and work occurred during the stance phase, whereas increased work was performed by the hip flexor muscles during the transition from stance to swing. In the frontal plane, increased hip and knee varus moments were present. These differences demonstrate that walking in high-heeled shoes alters lower-extremity joint kinetic function. Reduced effectiveness of the ankle plantar flexors during late stance results in a compensatory enhanced hip flexor "pull-off" that assists in limb advancement during the stance-to-swing transition. Larger muscle moments and increased work occur at the hip and knee, which may predispose long-term wearers of high-heeled shoes to musculoskeletal pain. PMID- 12533554 TI - Relationship between onychocryptosis and foot type and treatment with toe spacer. A preliminary investigation. AB - The relationship between onychocryptosis and foot type was investigated in a series of 512 patients. Of these patients, 124 had signs or a history of onychocryptosis. Among the nine foot types identified by digital and metatarsal formulas, the Greek index minus and squared index minus types showed the strongest association with onychocryptosis, which was present in more than one third of such feet. When anteroposterior radiographs of each type of foot were taken after binding the first and second toes together to simulate a tight shoe, the enlargement of bony structures of the second toe at the distal interphalangeal level in the Greek and squared index minus feet moved toward the distal enlargement of the distal phalanx of the first toe where the ingrowing occurs. Ten cases of stage I and four cases of stage II onychocryptosis were treated by placing a toe spacer between the first and second toes; all healed in about 3 weeks, suggesting that counterpressure of the second toe in tight shoes is a factor in the development of onychocryptosis. PMID- 12533555 TI - Comorbidities associated with diabetic foot complications among Asian Americans in southern California. AB - The comorbidities of diabetes mellitus were evaluated in an Asian American population with podiatric symptoms living in southern California. The three most common nonpedal complaints in men were blurred vision (73.6%), hypertension (64.1%), and erectile dysfunction (52.3%) and in women were blurred vision (84.5%), incontinence (71.5%), and low-back pain with radiculopathy-like symptoms (56.5%). The most significant finding was that only 3.2% of all patients had any previous knowledge or understanding of the risks of foot infection, ulceration, and amputation secondary to diabetes mellitus. The prevalence of diabetes mellitus in ethnic populations once considered practically exempt continues to rise steadily, and Asians living in the United States are becoming casualties of diabetes mellitus and its complications. PMID- 12533556 TI - Fall-contributing adverse effects of the most frequently prescribed drugs. AB - The 200 most frequently prescribed medications in 2000 were reviewed for adverse effects that have the potential to cause fall injuries. The actual number of different medications reviewed was 169 after eliminating duplicates due to listing of medications by both brand and generic names. Of these 169 medications, adverse effects of documented traumatic injuries and falls were reported for 9.5% (n = 16). Four hundred forty-eight adverse effects were identified and organized into 13 broad categories representing drug-induced changes in nervous, circulatory, and muscular systems. These changes were reported for 157 medications reviewed (92.9%) and could result in fall injuries. The accompanying list of medications can serve as a ready reference for podiatric physicians and other health-care professionals when monitoring and counseling patients regarding the potential for medication-induced fall injuries, which are especially common in the elderly population. PMID- 12533557 TI - Multiple schwannomas of the foot. Case report and strategy for treatment. AB - Determining the appropriate treatment of a benign tumor of a peripheral nerve in the foot and ankle region presents a clinical dilemma, as resection of the tumor will cause loss of nerve function and create the possibility of a painful neuroma. Several surgical solutions to this problem were used in the care of a patient who presented with painful bilateral Morton's neuromas and was found to have bilateral schwannomas on pathologic examination of the resected nerves. Subsequent evaluation for recurrent bilateral foot pain demonstrated multiple tumors along the tibial nerve in one foot. The patient also became aware of a painful mass on the dorsolateral aspect of one foot. Review of the treatment options for this patient with multiple schwannomas provides a framework for decision making in the care of the patient with benign neural tumors of the foot. PMID- 12533558 TI - Lower-extremity wounds inflicted by rats. PMID- 12533559 TI - Merton L. Root: 1922-2002. PMID- 12533560 TI - An approach to foot orthopedics. 1964. PMID- 12533562 TI - 2002 Podiatric Practice Survey. Statistical results. AB - This report presents the results of analyses of statistical data from 2,955 members of the American Podiatric Medical Association who responded to the 2002 Podiatric Practice Survey, conducted in April through May 2002. PMID- 12533564 TI - Introducing levels of evidence to the journal. PMID- 12533565 TI - The clinical importance of meniscal tears demonstrated by magnetic resonance imaging in osteoarthritis of the knee. AB - BACKGROUND: Meniscal tears are frequently found during magnetic resonance imaging of osteoarthritic knees. However, the prevalence and clinical relevance of these tears have not been determined. This study was designed to investigate the relationship between meniscal tears and osteoarthritis and between such tears and pain in patients with osteoarthritis. METHODS: Magnetic resonance imaging and plain radiography of the knee were performed in a group of 154 patients with clinical symptoms of knee osteoarthritis and a group of forty-nine age-matched asymptomatic controls. Pain scores (according to a 100-mm visual analog scale) and functional scores (according to the Western Ontario and McMaster University Osteoarthritis Index [WOMAC]) were determined for ninety-one of the patients with symptomatic osteoarthritis. Meniscal tears were defined as tears extending to an articular surface as seen on magnetic resonance imaging. RESULTS: A medial or lateral meniscal tear was a very common finding in the asymptomatic subjects (prevalence, 76%) but was more common in the patients with symptomatic osteoarthritis (91%) (p < 0.005). In the group with symptomatic osteoarthritis, a higher Kellgren-Lawrence radiographic grade was correlated with a higher frequency of meniscal tears (r = 0.26, p < 0.001), and men had a higher prevalence of meniscal tears than did women (p < 0.01). However, there was no significant difference with regard to the pain or WOMAC score between the patients with and those without a medial or lateral meniscal tear in the osteoarthritic group (p = 0.8 to 0.9 for all comparisons). The power of the study was 80% to detect a difference in the WOMAC scores of 15 points and a difference in the scores on the visual analog scale of 16 mm. CONCLUSIONS: Meniscal tears are highly prevalent in both asymptomatic and clinically osteoarthritic knees of older individuals. However, osteoarthritic knees with a meniscal tear are not more painful than those without a tear, and the meniscal tears do not affect functional status. These data do not support the routine use of magnetic resonance imaging for the evaluation and management of meniscal tears in patients with osteoarthritis of the knee. LEVEL OF EVIDENCE: Diagnostic study, Level I-1 (testing of previously developed diagnostic criteria in series of consecutive patients [with universally applied reference "gold" standard]). See p. 2 for complete description of levels of evidence. PMID- 12533566 TI - Effect of arthroscopic debridement for osteoarthritis of the knee on health related quality of life. AB - BACKGROUND: The available evidence supporting the use of arthroscopic debridement for the treatment of symptomatic osteoarthritis of the knee is largely retrospective and lacks validated health-related quality-of-life measures. The goal of the study was to prospectively assess a cohort of patients with osteoarthritis of the knee who were selected for arthroscopic debridement and determine which clinical criteria favor a sustained improvement in health-related quality of life after two years of follow-up. METHODS: One hundred and twenty-six patients with symptomatic primary osteoarthritis of the knee underwent arthroscopic debridement of the knee after failure of medical management. Two groups of surgeons (postgraduate fellows and attending staff) independently evaluated the patients preoperatively with use of a standardized assessment of clinical symptoms and signs and plain radiography. The intervention was arthroscopic debridement, which included resection of unstable chondral flaps and meniscal tears. Abrasion was not performed. Outcome measures included the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), a disease specific scale, and the Short Form-36 (SF-36), a generic outcome measure. RESULTS: Sixty-seven (53%) of the 126 patients were female, and the mean age (and standard deviation) was 61.7 +/- 8.6 years (range, forty-three to seventy-five years). The medial compartment was more frequently and severely involved, with 57% showing Grade-III or IV involvement, according to the system of Dougados et al. Seventy-nine patients (63%) had an unstable meniscal tear. Fifty-six patients (44%) were rated as having had a clinically important reduction in pain, as determined with the WOMAC pain scale, at two years after the arthroscopic debridement. Physicians were poor at predicting which patients would have improvement. The rate of accuracy of the fellows and staff was 54% and 59%, respectively, and their agreement was only slightly better than chance, with a kappa of 0.27 (95% confidence interval, 0.09 to 0.45). Only three variables were significantly associated with improvement: the presence of medial joint-line tenderness (p = 0.04), a positive Steinman test (p = 0.01), and the presence of an unstable meniscal tear at arthroscopy (p = 0.01). CONCLUSIONS: The prospectively evaluated quality-of-life benefit from arthroscopic debridement of the osteoarthritic knee is less than that reported in previous retrospective surveys on satisfaction. These results may serve as a baseline for comparison against more sophisticated procedures for resurfacing of the articular cartilage. Clinical variables were only partially helpful for predicting a successful result after arthroscopic debridement, and a search for other biologic markers (such as synovial fluid) may be of benefit. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See p. 2 for complete description of levels of evidence. PMID- 12533567 TI - Incidence rates of dislocation, pulmonary embolism, and deep infection during the first six months after elective total hip replacement. AB - BACKGROUND: The lengths of acute hospital stays following total hip replacement have diminished substantially in recent years. As a result, a greater proportion of complications occurs following discharge. Data on the incidence trends of major complications of total hip replacement would facilitate recognition and management of these adverse events. METHODS: We used Medicare claims data on beneficiaries sixty-five years and older who had had elective, primary total hip replacement for a reason other than a fracture (58,521 patients) or had had revision total hip replacement (12,956 patients) between July 1, 1995, and June 30, 1996. We calculated incidence rates of dislocation, pulmonary embolism, and deep hip infection per 10,000 person-weeks for four time-periods following the admission for the surgery (during the index hospitalization, from discharge to four weeks postoperatively, from five to thirteen weeks postoperatively, and from fourteen to twenty-six weeks postoperatively). We then used life-table methods to estimate the cumulative incidence of each complication over the first six postoperative months. RESULTS: Of the patients who had had a primary total hip replacement, 3.9% had a dislocation, 0.9% had a pulmonary embolism, and 0.2% had a deep infection in the first twenty-six postoperative weeks. In the revision total hip replacement cohort, the proportions with dislocation, pulmonary embolism, and deep infection were 14.4%, 0.8%, and 1.1%, respectively. The rates of these adverse outcomes were highest during the index hospitalization, diminished considerably in the period from discharge to four weeks postoperatively, and continued to drop in the periods from five to thirteen and fourteen to twenty-six weeks postoperatively. CONCLUSIONS: The incidence rates of dislocation, pulmonary embolism, and deep infection are highest immediately after total hip replacement, but they continue to be elevated throughout the first three postoperative months. With the lengths of hospital stays continuing to diminish, an increasing proportion of complications will occur in outpatients. These findings provide a basis for developing strategies to prevent these complications in the postdischarge management of patients who have had elective total hip replacement. LEVEL OF EVIDENCE: Prognostic study, Level II-1 (retrospective study). See p. 2 for complete description of levels of evidence. PMID- 12533568 TI - Rates and outcomes of primary and revision total hip replacement in the United States medicare population. AB - BACKGROUND: Information on the epidemiology of primary total hip replacement is limited, and we are not aware of any reports on the epidemiology of revision total hip replacement. The objective of this study was to characterize the rates and immediate postoperative outcomes of primary and revision total hip replacement in persons sixty-five years of age and older residing in the United States. METHODS: We used Medicare claims submitted by hospitals, physicians, and outpatient facilities between July 1, 1995, and June 30, 1996, to identify individuals who had undergone elective primary total hip replacement for a reason other than a fracture (61,568 patients) or had had revision total hip replacement (13,483 patients). Annual incidence rates of primary and revision total hip replacement were calculated, and multivariate modeling was used to evaluate the association between patient characteristics and surgical rates. The rates of occurrence of five complications within ninety days postoperatively were also evaluated, and relationships between those outcomes and patient characteristics were assessed with use of multivariate models adjusted for hospital and surgeon volume. RESULTS: The rates of primary total hip replacement were three to six times higher than the rates of revision total hip replacement. Women had higher rates than men, and whites had higher rates than blacks. The rates of primary and revision total hip replacement increased with age until the age of seventy-five to seventy-nine years and then declined. The rates of complications occurring within ninety days after primary total hip replacement were 1.0% for mortality, 0.9% for pulmonary embolus, 0.2% for wound infection, 4.6% for hospital readmission, and 3.1% for hip dislocation. The rates after revision total hip replacement were 2.6%, 0.8%, 0.95%, 10.0%, and 8.4%, respectively. Factors associated with an increased risk of an adverse outcome included increased age, gender (men were at higher risk than women), race (blacks were at higher risk than whites), a medical comorbidity, and a low income. CONCLUSIONS: Analysis of United States Medicare population data showed that the rates of total hip replacement increased with age up to the age of seventy-five to seventy-nine years and that blacks had a significantly lower rate of total hip replacement than whites. The overall rates of adverse outcomes were relatively low, but they were significantly higher after revision than after primary total hip replacement. LEVEL OF EVIDENCE: Prognostic study, Level II-1 (retrospective study). See p. 2 for complete description of levels of evidence. PMID- 12533569 TI - Open reduction and internal fixation of delayed unions and nonunions of fractures of the distal part of the humerus. AB - BACKGROUND: The purpose of the present retrospective study was to evaluate the results of open reduction and internal fixation of delayed unions and nonunions of fractures of the distal part of the humerus. METHODS: Between 1976 and 2001, fifty-two patients with a delayed union (thirteen patients) or nonunion (thirty nine patients) of the distal part of the humerus were treated with open reduction and internal fixation along with selective elbow joint arthrolysis and bone grafting. The average time to presentation was eighteen months (range, two to 192 months) after the injury. Thirty-nine of the fifty-two patients had undergone an average of 1.6 previous operations. There were twenty-seven supracondylar, six transcondylar, thirteen intercondylar, two lateral condylar, and four medial condylar delayed unions or nonunions. The average duration of follow-up was thirty-three months (range, three to 198 months). RESULTS: Fifty-one of the fifty two patients had healing of the delayed union or nonunion after the index operation; the average time to union was six months (range, two to twenty-four months). The average range of elbow motion increased from 71 degrees preoperatively to 94 degrees postoperatively. Complications included two superficial infections, two deep infections, and five cases of ulnar neuropathy. Fifteen patients (29%) needed additional surgery after the index procedure. Specifically, seven patients underwent removal of prominent hardware; six underwent hardware removal along with excision of heterotopic bone, ulnar neurolysis, and/or manipulation under anesthesia; one underwent irrigation and debridement; and one underwent compartment release. CONCLUSIONS: Open reduction through an extensile exposure and rigid internal fixation consistently results in healing of a delayed union or nonunion of the distal part of the humerus. An improved range of motion of the elbow can be achieved by securing the site of the nonunion and performing aggressive elbow joint arthrolysis and soft-tissue releases in patients with severe contractures. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See p. 2 for complete description of levels of evidence. PMID- 12533570 TI - Isolated revision of the patellar component in total knee arthroplasty. AB - BACKGROUND: Problems with the patellofemoral articulation are the most common causes of failure after total knee arthroplasty. However, there are few reports describing outcomes following isolated revision of the patellar component. METHODS: Forty knees with a Miller-Galante I prosthesis underwent isolated patellar revision (with or without lateral retinacular release). The Hospital for Special Surgery knee scores were collected prospectively, and radiographs made preoperatively and at the time of the final follow-up were analyzed with respect to alignment, component position, and patellar tracking. Particular attention was given to patients who had a reoperation or repeat revision and who had clinical or radiographic evidence of failure of the patellar revision. RESULTS: At a mean follow-up of sixty-two months, fifteen (38%) of the forty knees that had had an isolated revision of the patellar component failed a second time. Eight of them required a total of twelve additional operations at a mean of forty-nine months after the patellar revision. Three of the failures were severe enough to require revision of two or more of the components. Of the twenty-five knees that had not failed, the average Hospital for Special Surgery knee score at the time of the final follow-up was 87 points. Of the seven knees that did not undergo reoperation but were deemed to be failures on the basis of the patients' symptoms, the average Hospital for Special Surgery knee score at the time of the final follow-up was 72 points. CONCLUSIONS: Isolated patellar revision, with or without concurrent lateral retinacular release, was associated with a high rate of reoperation and a relatively low rate of success. Elements of the implant design and component alignment contributed to the patellar component failure; both should be scrutinized carefully in patients who are seen with this problem, prior to proceeding with isolated revision of the patellar component of a total knee arthroplasty. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See p. 2 for complete description of levels of evidence. PMID- 12533571 TI - Effect of selective capsulorrhaphy on the passive range of motion of the glenohumeral joint. AB - BACKGROUND: Capsulorrhaphy of the glenohumeral joint is a common surgical procedure for the treatment of instability caused by increased capsular laxity. The effect of capsulorrhaphy on the range of motion of the shoulder is poorly understood. METHODS: We simulated localized capsular contractures by selective capsular plications in eight human cadaveric shoulders and studied the effect of such plications on the passive range of glenohumeral abduction, flexion, and external and internal rotation in different degrees of abduction. A 0.5 or 1-N-m torque was applied to the humerus, and the range of glenohumeral motion was measured with electronic goniometers in three planes and compared with those of the intact shoulder. RESULTS: Anterosuperior capsular plication most markedly affected external rotation of the adducted arm, decreasing it by a mean of 30.1 degrees (p < 0.0001). Anteroinferior plication significantly reduced abduction by a mean of 19.4 degrees (p < 0.0001) and external rotation by a mean of 20.6 degrees (p = 0.0046). Posterosuperior plication mostly limited internal rotation of the adducted arm (mean decrease, 16.1 degrees, p = 0.0045). On the average, total anterior and total posterior plication each limited flexion by approximately 20 degrees (p = 0.005) and abduction by >or=15 degrees (p < 0.005), whereas total anterior plication limited external rotation by >30 degrees (p 20 degrees (p < 0.0001). Total inferior capsular plication restricted abduction (by a mean of 27.7 degrees, p = 0.0001), flexion, and rotation. Total superior plication restricted external rotation and flexion. CONCLUSIONS AND CLINICAL RELEVANCE: Localized plications of the glenohumeral joint capsule lead to predictable patterns of loss of glenohumeral mobility. If plication is planned, losses of movement can be anticipated. The findings of this study may assist surgeons in identifying the parts of the capsule that are contracted and that may need lengthening. PMID- 12533572 TI - Fate of the unrevised all-polyethylene patellar component in revision total knee arthroplasty. AB - BACKGROUND: During revision total knee arthroplasty, the alternatives for addressing the well-positioned and secure all-polyethylene patellar component with minimal or no wear include retention, revision, and resection. The purpose of the present study was to determine the intermediate-term fate of all polyethylene patellar components that were left in situ at the time of revision tibiofemoral arthroplasty. METHODS: We retrospectively reviewed the results of 202 revision total knee arthroplasties, performed at four centers, in which a well-fixed, well-positioned all-polyethylene patellar component with minimal wear was retained. Clinical evaluation was performed to identify the presence of anterior knee symptoms or patellar failures, and the Knee Society clinical scores were recorded. RESULTS: Sixty-eight percent of the revision tibiofemoral components were from a different manufacturer than the retained patellar component. Twenty-one patients (twenty-one knees; 10%) reported anterior knee pain at a mean duration of follow-up of seven years (range, two to fourteen years). Eleven of the twenty-one knees with anterior knee pain had evidence of component loosening, excessive wear, or delamination either on follow-up radiographs or on direct inspection at the time of reoperation. In the other ten knees, the findings were attributable to osseous impingement on the femoral component or soft-tissue dysfunction (including soft-tissue impingement, subluxation, and scarring). Seventeen of the twenty-one patients with anterior knee pain underwent additional surgery to address the source of the pain. Failures due to loosening or wear occurred only in patients in whom the patellar component had been sterilized with gamma irradiation in air; these failures occurred at a mean 7.3 years after revision (eleven years after primary total knee arthroplasty). The likelihood of patellar component failure was significantly greater when the component had been gamma irradiated in air than when it had been sterilized with another method (p = 0.0008). The average knee score increased from 42 to 86 points, and the average function score increased from 46 to 70 points. CONCLUSIONS: Retaining a well-positioned, stable all polyethylene patellar component at the time of revision tibiofemoral arthroplasty can be successful, provided that the polyethylene has not oxidized. Manufacturing mismatch is acceptable with most contemporary designs provided that the patellar component articulates appropriately with the femoral implant. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See p. 2 for complete description of levels of evidence. PMID- 12533573 TI - The Lapidus procedure as salvage after failed surgical treatment of hallux valgus: a prospective cohort study. AB - BACKGROUND: Recurrent hallux valgus is a relatively common, yet challenging, condition for both the patient and the surgeon. The literature on the treatment of recurrent hallux valgus is sparse. The purpose of this study was to evaluate prospectively the functional outcome and patient satisfaction following the Lapidus procedure for the treatment of recurrent hallux valgus deformity. METHODS: Twenty-four patients with a total of twenty-six symptomatic recurrences of hallux valgus after previous procedures for treatment of the deformity were included in the study. Exclusion criteria included prior fusion procedures on the foot or ankle, a previous Keller or Mayo procedure, insulin-dependent diabetes, peripheral vascular disease, or peripheral neuropathy. A visual analog pain scale and the American Orthopaedic Foot and Ankle Society Hallux Metatarsophalangeal Interphalangeal Scale were administered preoperatively, at six months postoperatively, and yearly thereafter. Weight-bearing radiographs were also made preoperatively; at six weeks, three months, six months, and one year postoperatively; and yearly thereafter. Patient satisfaction was assessed at the latest follow-up evaluation. RESULTS: At twenty-four months, the mean score according to the American Orthopaedic Foot and Ankle Society (AOFAS) Hallux Metatarsophalangeal-Interphalangeal Scale had increased from 47.6 to 87.9 points, the mean score according to the visual analog pain scale had improved from 6.2 to 1.4, the mean hallux valgus angle had improved from 37.1 degrees to 17.1 degrees, and the mean intermetatarsal angle had improved from 18 degrees to 8.6 degrees. The patients were very satisfied after 77% of the twenty-six procedures, satisfied after 4%, and somewhat satisfied after 19%; no patient was dissatisfied. There were no cases of hallux varus. Complications included three nonunions, all of which occurred in smokers, and two superficial wound infections. CONCLUSION: In appropriately selected patients, the Lapidus procedure is a reliable and effective operation after failed surgical treatment of hallux valgus. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See p. 2 for complete description of levels of evidence. PMID- 12533574 TI - Clinical features of the different types of SLAP lesions: an analysis of one hundred and thirty-nine cases. AB - BACKGROUND: Previous studies have suggested that SLAP (superior labrum anterior posterior) lesions are a distinct clinical entity. The goals of this study were to define the prevalence, associated pathological findings, and clinical features of the different types of SLAP lesions with use of a common classification system. METHODS: Five hundred and forty-four patients undergoing shoulder arthroscopy for a variety of diagnoses were prospectively included in this consecutive case series. SLAP lesions were grouped with use of the Snyder classification. Demographic data, clinical data, and arthroscopic findings in the groups with SLAP lesions were compared with those in a control group with no SLAP lesion. RESULTS: Of 544 shoulder arthroscopy procedures, 139 (26%) demonstrated a SLAP lesion. One hundred and three (74%) of the SLAP lesions were Type I, twenty nine (21%) were Type II, one (0.7%) was Type III, and six (4%) were Type IV. Most (123) of the SLAP lesions were found to be associated with other intra-articular lesions. Multivariate analysis revealed that a positive Speed test and a supraspinatus tear were significantly associated with Type-I lesions (p = 0.012 and p = 0.001, respectively). The findings associated with Type-II lesions differed according to the patient's age: Type-II lesions in patients who were forty years of age or younger were associated only with a Bankart lesion, whereas those in patients older than forty years of age were associated with a supraspinatus tear and osteoarthritis of the humeral head. Type-III and Type-IV lesions were associated with a high-demand occupation and a Bankart lesion. CONCLUSIONS: This study demonstrated that the prevalence, associated pathological findings, and clinical features of the different types of SLAP lesions vary with the patient population that is studied. Also, the clinical features and pathological findings associated with the different types of SLAP lesions often overlap. Isolated SLAP lesions with no associated pathological findings are uncommon, and care must be taken when ascribing symptoms to a SLAP lesion when other lesions are present. LEVEL OF EVIDENCE: Diagnostic study, Level IV-1 (case control study). See p. 2 for complete description of levels of evidence. PMID- 12533575 TI - Central placement of the screw in simulated fractures of the scaphoid waist: a biomechanical study. AB - BACKGROUND: Recent reports on internal fixation of acute fractures of the scaphoid waist have demonstrated higher rates of central placement of the screw when cannulated screws were used than when noncannulated screws were used. This cadaveric study was designed to determine whether central placement in the proximal fragment of the scaphoid offers a biomechanical advantage. METHODS: Eleven matched pairs of scaphoids were removed from fresh cadaveric wrists. Each scaphoid was placed in a custom manufactured jig that was used to create reproducible central and eccentric positioning of the guidewire. Then a linear osteotomy was made followed by placement of a Herbert-Whipple cannulated screw to fix the osteotomy. The specimen was then potted in a holder with use of polymethylmethacrylate with a Kirschner wire passed through the proximal end of the scaphoid and placed into a fixture with a pneumatically driven plunger resting on the surface of the distal pole. The load acting through the plunger was measured with use of a load-cell, and its excursion was measured with use of a linear variable differential transformer. Stiffness, load at 2 mm of displacement, load at failure, and mechanism of failure were measured, and the two groups were compared with regard to stiffness and strength. RESULTS: Central placement of the screw in the proximal fragment of the scaphoid had superior results compared with those after eccentric positioning of the screw. Fixation with central placement of the screw demonstrated 43% greater stiffness (12.7 N per mm compared with 8.9 N per mm; p < 0.01), 113% greater load at 2 mm of displacement (126 N compared with 59.1 N; p < 0.01), and 39% greater load at failure (712 N compared with 513 N; p > 0.05). CONCLUSIONS AND CLINICAL RELEVANCE: Central placement of the screw in the proximal fragment of the scaphoid offers a biomechanical advantage in the internal fixation of an osteotomy of the scaphoid waist. Clinical efforts and techniques that facilitate central placement of the screw in the fixation of fractures of the scaphoid waist should be encouraged. PMID- 12533576 TI - Medial opening-wedge high tibial osteotomy with use of porous hydroxyapatite to treat medial compartment osteoarthritis of the knee. AB - BACKGROUND: The aims of this study were to investigate the results of opening wedge high tibial valgus osteotomy in patients with medial compartment osteoarthritis of the knee and to examine the usefulness of hydroxyapatite wedges as the supporting material. METHODS: Medial opening-wedge osteotomy was performed in twenty-one osteoarthritic knees in eighteen patients who had a mean age of 66.6 years. The mean duration of follow-up was 78.6 months. A medial transverse osteotomy was performed proximal to the tibial tuberosity, with the most lateral 10% of the tibia left intact. The medial side of the osteotomy site was opened to the desired angle of correction. Two hydroxyapatite wedges of the same size (5.0, 7.5, or 10.0 mm) were inserted into the opened osteotomy site along with bone grafts, and the fragments were fixed with two plates. The angle of correction could be adjusted by altering the direction of wedge insertion. The goal was to achieve a final standing alignment of 10 degrees of anatomical valgus angulation. RESULTS: All patients had pain relief and improvement in walking ability after the osteotomy. The mean knee and function scores of the American Knee Society were 60.2 +/- 5.3 and 48.1 +/- 10.4 points, respectively, before the osteotomy and 94.3 +/- 7.3 and 93.1 +/- 9.8 points, respectively, at the time of the final follow-up. Limb alignment, expressed as the standing femorotibial angle, was corrected from 180 degrees +/- 2.9 degrees preoperatively to 169.7 degrees +/- 3.7 degrees (10.3 degrees of anatomical valgus angulation) at the time of the latest follow-up. There were no cases of recurrence of varus deformity or collapse of the hydroxyapatite wedges. CONCLUSIONS: After a mean duration of follow-up of 6.6 years, we found that the medial opening-wedge osteotomy of the proximal part of the tibia provided satisfactory clinical results for patients with osteoarthritis of the knee. Use of the porous hydroxyapatite wedges resulted in no collapse or subsidence at the osteotomy site. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See p. 2 for complete description of levels of evidence. PMID- 12533577 TI - Sustained spinal cord compression: part I: time-dependent effect on long-term pathophysiology. AB - BACKGROUND: The objective of this study is to determine whether there is a relationship between the duration of sustained spinal cord compression and the extent of spinal cord injury and the capacity for functional recovery after decompression. METHODS: Sixteen dogs underwent sustained spinal cord compression for thirty or 180 minutes. The cords were compressed with use of a loading device with a hydraulic piston. A pressure transducer was attached to the surface of the piston, which transmitted real-time spinal cord interface pressures to a data acquisition system. Somatosensory evoked potentials were monitored during a sixty minute recovery period as well as at twenty-eight days after the injury. Functional motor recovery was judged throughout a twenty-six-day period after the injury with use of a battery of motor tasks. The volume of the lesion and damage to the tissue were assessed with both magnetic resonance imaging and histological analysis. RESULTS: Sustained spinal cord compression was associated with a gradual decline in interface pressure. Despite this, there was continuous decline in the amplitude of the somatosensory evoked potentials, which did not return until the cord was decompressed. Within one hour after the decompression, the dogs in the thirty-minute-compression group had recovery of somatosensory evoked potentials, but no animal had such recovery in the 180-minute group. Recovery of the somatosensory evoked potentials in the thirty-minute group was sustained over the twenty-eight days after the injury. Motor tests demonstrated rapid recovery of hindlimb motor function in the thirty-minute group, but there was considerable impairment in the 180-minute group. Within two weeks after the injury, balance, cadence, stair-climbing, and the ability to walk up an inclined plane were significantly better in the thirty-minute group than in the 180-minute group. The longer duration of compression produced lesions of significantly greater volume, which corresponded to the long-term functional outcome. CONCLUSIONS: The relatively rapid viscoelastic relaxation of the spinal cord during the early phase of sustained cord compression suggests that there are mechanisms of secondary injury that are linked to tissue displacement. Longer periods of displacement allow propagation of the secondary injury process, resulting in a lack of recovery of somatosensory evoked potentials, limited functional recovery, and more extensive tissue damage. PMID- 12533578 TI - Sustained spinal cord compression: part II: effect of methylprednisolone on regional blood flow and recovery of somatosensory evoked potentials. AB - BACKGROUND: The efficacy of methylprednisolone in the treatment of traumatic spinal cord injury is controversial. We examined the effect of methylprednisolone on regional spinal cord blood flow and attempted to determine whether recovery of electrophysiological function is dependent on reperfusion, either during sustained spinal cord compression or after decompression. METHODS: The effects of methylprednisolone therapy on recovery of somatosensory evoked potentials and on spinal cord blood flow were examined in a canine model of dynamic spinal cord compression. Methylprednisolone (30 mg/kg intravenous loading dose followed by 5.4 mg/kg/hr intravenous infusion) or saline solution was administered to thirty six beagles (eighteen in each group) five minutes after cessation of dynamic spinal cord compression and loss of all somatosensory evoked potentials. After ninety minutes of sustained compression, the spinal cords were decompressed. Somatosensory evoked potentials and spinal cord blood flow were evaluated throughout the period of sustained compression and for three hours after decompression. RESULTS: Seven dogs treated with methylprednisolone and none treated with saline solution recovered measurable somatosensory evoked potentials during sustained compression. After decompression, three more dogs treated with methylprednisolone and seven dogs treated with saline solution recovered somatosensory evoked potentials. Four dogs treated with methylprednisolone lost their previously measurable somatosensory evoked potentials. In the methylprednisolone group, spinal cord blood flow was significantly higher (p < 0.05) in the dogs that had recovered somatosensory evoked potentials than it was in the dogs that had not. Reperfusion blood flow was significantly higher (p < 0.05) in the saline-solution group than it was in the methylprednisolone group. Spinal cord blood flow in the saline-solution group returned to baseline levels within five minutes after decompression. It did not return to baseline levels in the dogs treated with methylprednisolone. CONCLUSIONS: The methylprednisolone administered in this study did not provide a large or significant lasting benefit with regard to neurological preservation or restoration. Methylprednisolone may reduce regional spinal cord blood flow through mechanisms affecting normal autoregulatory blood-flow function. PMID- 12533579 TI - Clinical outcomes after lumbar discectomy for sciatica: the effects of fragment type and anular competence. AB - BACKGROUND: The surgical treatment of sciatica with discectomy is ineffective in a sizable percentage of patients, and reherniation occurs after 5% to 15% of such procedures. The purpose of the present study was to determine if competence of the disc anulus and the type of herniation could be used to predict postoperative clinical outcomes following lumbar discectomy. METHODS: A prospective observational study of 187 consecutive patients undergoing single-level primary lumbar discectomy was conducted. A single surgeon performed all of the procedures, and an independent examiner evaluated 180 of the patients clinically at a minimum of two and a median of six years after surgery. The extent of anular deficiency and the presence of disc fragments were determined. On the basis of these intraoperative findings, disc herniations were classified into four categories: (1) Fragment-Fissure herniations (eighty-nine patients), (2) Fragment Defect herniations (thirty-three patients), (3) Fragment-Contained herniations (forty-two patients), and (4) No Fragment-Contained herniations (sixteen patients). The effects of disc herniation morphology and preoperative variables on subsequent clinical outcome were determined with the Student t test for continuous variables and chi-square analysis for categorical variables. RESULTS: Patients in the Fragment-Fissure group, who had disc fragments and a small anular defect, had the best overall outcomes and the lowest rates of reherniation (1%) and reoperation (1%). Patients in the Fragment-Contained group had a 10% rate of reherniation and a 5% rate of reoperation. Patients in the Fragment-Defect group, who had extruded fragments and massive posterior anular loss, had a 27% rate of reherniation and a 21% rate of reoperation. Patients in the No Fragment-Contained group did poorly: 38% had recurrent or persistent sciatica, and the standard outcomes scores were less improved compared with those in the other groups (p < 0.001). CONCLUSION: Intraoperative findings, as described in the present study, were more clearly associated with outcomes than were demographic, socioeconomic, or clinical variables. The degree of anular competence after discectomy and the type of herniation appear to have value for the prediction of the recurrence of sciatica, reoperation, and clinical outcome following lumbar discectomy. LEVEL OF EVIDENCE: Prognostic study, Level I-1 (prospective study). See p. 2 for complete description of levels of evidence. PMID- 12533580 TI - Primary total hip arthroplasty with a second-generation cementless total hip prosthesis in patients younger than fifty years of age. AB - BACKGROUND: The early experience with cementless total hip replacement led to design modifications to increase the initial press-fit and stability of the femoral component, to reduce the diameter of the femoral head, and to improve the acetabular component, including the locking mechanism of the shell and the quality of the polyethylene liner. We performed a prospective study to assess the results in young patients who had been followed for a minimum of eight years after treatment with a primary total hip arthroplasty with a second-generation cementless prosthesis. METHODS: Eighty patients (118 hips) were included in the study. There were fifty-eight men and twenty-two women; the mean age at the time of the operation was 46.8 years (range, twenty-one to forty-nine years). We used a cementless Duraloc series-100 or 1200 acetabular component with or without screw fixation, a polyethylene liner with a 22-mm inner diameter, and a cementless Profile femoral component in all hips. The average duration of follow up was 9.8 years (range, eight to eleven years). Clinical follow-up (with use of the Harris hip score) and radiographic follow-up were performed at six weeks; at three, six, and twelve months; and yearly thereafter. Linear and volumetric wear were measured, and bone-remodeling and osteolysis were assessed. RESULTS: The average Harris hip score improved from 48.8 points preoperatively to 92 points at the final follow-up examination. The prevalence of transitory thigh pain was 10% (twelve of 118 hips). There was no aseptic loosening. One hip was revised because of recurrent dislocation. The average amount of linear wear was 1.18 mm, and the average wear rate was 0.12 mm/yr. Fourteen hips (12%) had osteolysis in the calcar femorale and eleven hips (9%) had acetabular osteolysis, but all of the osteolytic lesions were <1 cm (2). CONCLUSIONS: The mechanical fixation of the anatomic fit cementless Profile stem was excellent in this study of young patients who were particularly difficult to treat because of a high prevalence of osteonecrosis and developmental dysplasia. Although there was no aseptic loosening of the components and a relatively low prevalence of osteolysis, there was a high rate of linear wear of the polyethylene liner. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). See p. 2 for complete description of levels of evidence. PMID- 12533581 TI - Osteoporosis and anterior femoral notching in periprosthetic supracondylar femoral fractures: a biomechanical analysis. AB - BACKGROUND: This biomechanical study was designed to evaluate the predictive ability of dual-energy x-ray absorptiometry, cortical bone geometry as determined with computed tomography, and radiography in the assessment of torsional load to failure in femora with and without notching. METHODS: Thirteen matched pairs of cadaveric femora were randomized into two groups: a notched group, which consisted of femora with a 3-mm anterior cortical defect, and an unnotched group of controls. Each pair then underwent torsional load to failure. The ability of a number of measures to predict femoral torsional load to failure was assessed with use of regression analysis. These measures included dual-energy x-ray absorptiometry scans of the proximal and the distal part of the femur, geometric measures of both anterior and posterior cortical thickness as well as the polar moment of inertia of the distal part of the femur as calculated on computed tomography scans, and the Singh osteoporosis index as determined on radiographs. RESULTS: The torsional load to failure averaged 98.9 N-m for the notched femora and 143.9 N-m for the controls; the difference was significant (p < 0.01). Although several variables correlated with torsional load to failure, distal femoral bone-mineral density demonstrated the highest significant correlation (r = 0.85; p < 0.001). Moreover, multiple regression analysis showed that a combination of distal femoral bone-mineral density and polar moment of inertia calculated with the posterior cortical thickness (adjusted r (2) = 0.79; p < 0.001) had the strongest prediction of torsional load to failure in the notched group. The addition of other measures of cortical bone geometry, proximal femoral bone-mineral density, or radiographic evidence of osteopenia did not significantly increase the model's predictive ability. CONCLUSIONS: Femoral notching significantly decreases distal femoral torsional load to failure and is best predicted by a combination of the measures of distal femoral bone-mineral density and polar moment of inertia. Together, these values account for the amount of bone mass present and the stability provided by the cortical shell architecture. PMID- 12533582 TI - Spontaneous decrease in focal osteolysis in the unrevised femur after revision of a contralateral bipolar hemiarthroplasty: a case report. PMID- 12533583 TI - Chronic hip instability as a cause of autonomic dysreflexia: successful management by resection arthroplasty: a case report. PMID- 12533584 TI - Economic evaluation in orthopaedics. PMID- 12533586 TI - Evaluation and treatment of the arthritic knee. PMID- 12533587 TI - Barriers to full-text publication following presentation of abstracts at annual orthopaedic meetings. AB - BACKGROUND: Oral presentations at national and international meetings offer an excellent forum for the dissemination of current research findings. However, publication rates of full-text articles after presentation of abstracts at international meetings have ranged from 11% to 78%, which suggests that at least 32% of the abstracts presented are never published as complete articles in peer reviewed journals. In an effort to identify the reasons that surgeons had not had a paper published following presentation of their work at an international orthopaedic meeting, we conducted a survey of a cross section of authors of orthopaedic papers presented at a national meeting. METHODS: We retrieved all abstracts from the 1996 scientific program of the sixty-third Annual Meeting of the American Academy of Orthopaedic Surgeons. A computerized Medline and PubMed search established whether the abstract had been subsequently published as a full text article. The authors of the abstracts that had not been subsequently published were surveyed to identify the reasons for the failure to publish. RESULTS: A total of 465 abstracts were presented at the sixty-third Annual Meeting of the American Academy of Orthopaedic Surgeons in 1996. We surveyed the authors of 306 abstracts for which we were unable to locate a subsequent full text publication on Medline. One hundred and ninety-nine investigators (65%) responded to the questionnaire. At the time of the survey, seventy-two manuscripts had been published, thirty-two had been submitted and rejected, fourteen were under consideration by journals, seven had been accepted for publication or were in press, and three were not recalled by the investigator. In addition, seventy-one abstracts (35.7%) of the 199 had not been submitted for publication. The authors of those abstracts were asked to indicate one or more reasons why they had not submitted a manuscript for publication. Thirty-three investigators (46.5%) indicated that they lacked sufficient time for research activities, twenty-two (31.0%) reported that the study presented at the meeting in 1996 was still in progress, fourteen (19.7%) believed that the responsibility for writing the manuscript belonged to someone else, and twelve (16.9%) reported that difficulties with co-authors who would not participate had impeded the completion of the manuscript. Nine investigators (12.7%) responded that the pursuit of publication was a low priority. CONCLUSIONS: In a survey of investigators who had not had a full-text article published after presenting the abstract at a national meeting, we found that the failure to publish was due to one of three main reasons: (1) they did not have enough time to prepare a manuscript for publication (the reason most frequently given); (2) almost one third of the studies that had not been submitted for publication were ongoing; and (3) relationships with co-authors sometimes presented a barrier to final publication. Thorough preparation before the study and the establishment of stricter guidelines to limit the presentation of preliminary data at national and international meetings may improve publication rates. PMID- 12533588 TI - Placement of the posteromedial portal in posterior ankle arthroscopy. PMID- 12533589 TI - The orthopaedic profession and the orthopaedic industry: partners in quality patient care. PMID- 12533590 TI - Objectivity and worldly wisdom: missing ingredients in orthopaedic training and practice. PMID- 12533591 TI - Patterns of coronoid fractures and their treatment. PMID- 12533592 TI - Selection of orthopaedic trainees for specialist registrar posts in the United kingdom. PMID- 12533593 TI - Medical education and corporate sponsorship. PMID- 12533594 TI - What's new in shoulder and elbow surgery. PMID- 12533595 TI - Persistent changes in spontaneous firing of Purkinje neurons triggered by the nitric oxide signaling cascade. AB - Many types of neurons fire spontaneously because of the activity of pacemaking ion channels. Although endogenous firing can serve as a persistent signal to downstream targets, little attention has been paid to factors that might modulate such intrinsic electrical activity. We tested for modulation of spontaneous firing of Purkinje neurons in cerebellar slices under conditions in which principal synaptic inputs were blocked. Loose-patch recordings from single neurons show that sustained (>40 min) increases in the spontaneous firing rate can be triggered by activation of the nitric oxide-cGMP signaling pathway. Inhibitors of soluble guanylate cyclase and protein kinase G block this modulation. Increases in firing rate are also observed after stimulation of parallel fibers but not in response to basket cell activity. These findings elucidate a novel role for the nitric oxide-cGMP signaling cascade in the brain. This mechanism could permit long-term adjustments in the baseline firing rate of endogenously active neurons in response to changes in afferent activity. PMID- 12533596 TI - Synaptic activity of the AFD neuron in Caenorhabditis elegans correlates with thermotactic memory. AB - Thermotactic behavior in Caenorhabditis elegans is sensitive to both a worm's ambient temperature (T(amb)) and its memory of the temperature of its cultivation (T(cult)). The AFD neuron is part of a neural circuit that underlies thermotactic behavior. By monitoring the fluorescence of pH-sensitive green fluorescent protein localized to synaptic vesicles, we measured the rate of the synaptic release of AFD in worms cultivated at temperatures between 15 and 25 degrees C, and subjected to fixed, ambient temperatures in the same range. We found that the rate of AFD synaptic release is high if either T(amb) > T(cult) or T(amb) < T(cult), but AFD synaptic release is low if T(amb) congruent with T(cult). This suggests that AFD encodes a direct comparison between T(amb) and T(cult). PMID- 12533597 TI - Receptor occupancy limits synaptic depression at climbing fiber synapses. AB - Postsynaptic responses to presynaptic stimulation are used regularly to assess the amount of transmitter released from presynaptic release zones. At climbing fiber-to-Purkinje cell synapses, the number of vesicles released per active zone follows changes in release probability such that, normally, more than one vesicle is released per presynaptic action potential. This leads to high occupation of postsynaptic AMPA receptors by glutamate and thus may render the postsynaptic response relatively insensitive to changes in release. We find that paired-pulse depression of presynaptic release is not accurately represented by postsynaptic responses because of receptor saturation. By lowering vesicular glutamate concentrations or by using nonsaturated Bergmann glial AMPA receptors to monitor presynaptic release, we find that presynaptic depression of release is much greater than suggested previously. In addition, densely expressed glutamate transporters can shield Bergmann glial AMPA receptors and presynaptic metabotropic glutamate receptors from activation. PMID- 12533598 TI - Epsilon PKC is required for the induction of tolerance by ischemic and NMDA mediated preconditioning in the organotypic hippocampal slice. AB - Glutamate receptors and calcium have been implicated as triggering factors in the induction of tolerance by ischemic preconditioning (IPC) in the brain. However, little is known about the signal transduction pathway that ensues after the IPC induction pathway. The main goals of the present study were to determine whether NMDA induces preconditioning via a calcium pathway and promotes translocation of the protein kinase C epsilon (epsilonPKC) isozyme and whether this PKC isozyme is key in the IPC signal transduction pathway. We corroborate here that IPC and a sublethal dose of NMDA were neuroprotective, whereas blockade of NMDA receptors during IPC diminished IPC-induced neuroprotection. Calcium chelation blocked the protection afforded by both NMDA and ischemic preconditioning significantly, suggesting a significant role of calcium. Pharmacological preconditioning with the nonselective PKC isozyme activator phorbol myristate acetate could not emulate IPC, but blockade of PKC activation with chelerythrine during IPC blocked its neuroprotection. These results suggested that there might be a dual involvement of PKC isozymes during IPC. This was corroborated when neuroprotection was blocked when we inhibited epsilonPKC during IPC and NMDA preconditioning, and IPC neuroprotection was emulated with the activator of epsilonPKC. The possible correlation between NMDA, Ca2+, and epsilonPKC was found when we emulated IPC with the diacylglycerol analog oleoylacetyl glycerol, suggesting an indirect pathway by which Ca2+ could activate the calcium insensitive epsilonPKC isozyme. These results demonstrated that the epsilonPKC isozyme played a key role in both IPC- and NMDA-induced tolerance. PMID- 12533599 TI - A role for the juxtamembrane domain of beta-dystroglycan in agrin-induced acetylcholine receptor clustering. AB - Synaptic differentiation results from an exchange of informational molecules between synaptic partners during development. At the vertebrate neuromuscular junction, agrin is one molecule presented by the presynaptic motor neuron that plays an instructive role in postsynaptic differentiation of the muscle cell, most notably in aggregation of acetylcholine receptors (AChRs). Although agrin is the best-characterized synaptogenic molecule, its mechanism of action remains uncertain, but clearly, it requires the receptor tyrosine kinase MuSK (muscle specific kinase), the intracellular protein rapsyn, an Src-like kinase, and cytoskeletal components. In addition, the transmembrane protein dystroglycan interacts with the cytoskeleton and is implicated in agrin responsiveness. This alpha-beta heterodimer can bind agrin via its extracellular alpha subunit and associates with the membrane cytoskeleton via its beta subunit. In this study, we demonstrate that overexpression of the beta subunit of dystroglycan in cultured muscle cells inhibits agrin-induced AChR clustering. Deletion analysis and point mutagenesis demonstrate that the inhibition is mediated by an intracellular, juxtamembrane region composed of basic amino acids. Finally, the inhibition mediated by beta-dystroglycan extends to the minimal agrin fragment required for AChR clustering, suggesting that dystroglycan plays an important role in postsynaptic differentiation in response to agrin. PMID- 12533600 TI - Modulation of type 1 inositol (1,4,5)-trisphosphate receptor function by protein kinase a and protein phosphatase 1alpha. AB - Type 1 inositol (1,4,5)-trisphosphate receptors (InsP3R1s) play a major role in neuronal calcium (Ca2+) signaling. The InsP3R1s are phosphorylated by protein kinase A (PKA), but the functional consequences of InsP3R1 phosphorylation and the mechanisms that control the phosphorylated state of neuronal InsP3R1s are poorly understood. In a yeast two-hybrid screen of rat brain cDNA library with the InsP3R1-specific bait, we isolated the protein phosphatase 1alpha (PP1alpha). In biochemical experiments, we confirmed the specificity of the InsP3R1-PP1alpha association and immunoprecipitated the InsP3R1-PP1 complex from rat brain synaptosomes and from the neostriatal lysate. We also established that the association with PP1 facilitates dephosphorylation of PKA-phosphorylated InsP3R1 by the endogenous neostriatal PP1 and by the recombinant PP1alpaha. We demonstrated that exposure of neostriatal slices to 8-bromo-cAMP, dopamine, calyculin A, or cyclosporine A, but not to 10 nM okadaic acid, promotes the phosphorylation of neostriatal InsP3R1 by PKA in vivo. We discovered that PKA activates and PP1alpha inhibits the activity of recombinant InsP3R1 reconstituted into planar lipid bilayers. We found that phosphorylation of InsP3R1 by PKA induces at least a fourfold increase in the sensitivity of InsP3R1 to activation by InsP3 without shifting the peak of InsP3R1 bell-shaped Ca2+ dependence. Based on these data, we suggest that InsP3R1 may participate in cross talk between cAMP and Ca2+ signaling in the neostriatum and possibly in other regions of the brain. PMID- 12533601 TI - The second galanin receptor GalR2 plays a key role in neurite outgrowth from adult sensory neurons. AB - Expression of the neuropeptide galanin is markedly upregulated within the adult dorsal root ganglion (DRG) after peripheral nerve injury. We demonstrated previously that the rate of peripheral nerve regeneration is reduced in galanin knock-out mice, with similar deficits observed in neurite outgrowth from cultured mutant DRG neurons. Here, we show that the addition of galanin peptide significantly enhanced neurite outgrowth from wild-type sensory neurons and fully rescued the observed deficits in mutant cultures. Furthermore, neurite outgrowth in wild-type cultures was reduced to levels observed in the mutants by the addition of the galanin antagonist M35 [galanin(1-13)bradykinin(2-9)]. Study of the first galanin receptor (GalR1) knock-out animals demonstrated no differences in neurite outgrowth compared with wild-type animals. Similarly, use of a GalR1 specific antagonist had no effect on neuritogenesis. In contrast, use of a GalR2 specific agonist had equipotent effects on neuritogenesis to galanin peptide, and inhibition of PKC reduced neurite outgrowth from wild-type sensory neurons to that observed in galanin knock-out cultures. These results demonstrate that adult sensory neurons are dependent, in part, on galanin for neurite extension and that this crucial physiological process is mediated by activation of the GalR2 receptor in a PKC-dependent manner. PMID- 12533602 TI - Loss of kainate receptor-mediated heterosynaptic facilitation of mossy-fiber synapses in KA2-/- mice. AB - Multimeric assemblies of kainate (KA) receptor subunits form glutamate-gated ion channels that mediate EPSCs and function as presynaptic modulators of neurotransmitter release at some central synapses. The KA2 subunit is a likely constituent of many neuronal kainate receptors, because it is widely expressed in most neurons in the CNS. We have studied the effect of genetic ablation of this receptor subunit on synaptic transmission at the mossy-fiber-CA3 pyramidal cell synapse in hippocampal slices, where kainate receptors are localized to both presynaptic and postsynaptic sites. We found that both postsynaptic and presynaptic mossy-fiber kainate receptor function is altered in neurons from KA2 /- mice. The presynaptic facilitatory autoreceptor, which modulates glutamate release from mossy-fiber terminals, had a reduced affinity for exogenous agonists and synaptic glutamate. Although presynaptic facilitation attributable to homosynaptic glutamate release was normal at mossy-fiber synapses in KA2-/- neurons, heterosynaptic kainate receptor-mediated facilitation resulting from the spillover of glutamate from CA3 collateral synapses was absent. Consistent with a decrease in glutamate affinity of the receptor, the half-decay of the postsynaptic kainate-mediated EPSC was shorter in the knock-out mice. These results identify the KA2 subunit as a determinant of kainate receptor function at presynaptic and postsynaptic mossy-fiber kainate receptors. PMID- 12533603 TI - Connexin mediates gap junction-independent resistance to cellular injury. AB - Although gap junctions regulate essential processes during development and differentiation, the role of gap junctions in cell death is poorly understood. We demonstrate here that the forced expression of connexin 43 (Cx43), the main constituent of astrocytic gap junctions, protected against cell injury with a potency that was comparable with that from the expression of the proto-oncogene bcl2. The expression of two other members of the Cx family, Cx32 and Cx40, also increased the resistance to injury from exposures to calcium overload, oxidative stress, metabolic inhibition, tamoxifen, and UV irradiation, but not against staurosporine- and dexamethasone-mediated death. Surprisingly, the anti-death activity of connexin proteins was independent of gap junction channel function, because physical isolation or the pharmacological inhibition of coupling did not significantly increase cell death. Moreover, cells expressing nonfunctional mutant connexins also acquired a high resistance to injury. These observations identify Cx proteins as active players in cell survival. PMID- 12533604 TI - Bidirectional modulation of GABA release by presynaptic glutamate receptor 5 kainate receptors in the basolateral amygdala. AB - The activation of kainate receptors modulates GABAergic synaptic transmission, but the mechanisms are currently a matter of intense debate. In the basolateral amygdala (BLA), the glutamate receptor 5 (GluR5) subunit of kainate receptors is heavily expressed, and GluR5 antagonists block a novel form of synaptic plasticity; yet little is known about the role of GluR5-containing kainate receptors in the physiology of the amygdala. Here we show that GluR5 agonists bidirectionally modulate the strength of synaptic transmission from GABAergic interneurons to pyramidal cells in a concentration-dependent manner. Low concentrations of (RS)-S-amino-3-(3-hydroxy-5-tert-butylisoxazol-4-yl) (ATPA) (0.3 microm) or glutamate (5 microm) reduced the number of failures of GABAergic synaptic transmission and enhanced the frequency of miniature IPSCs (mIPSCs). High concentrations of ATPA (10 microm) or glutamate (200 microm) increased the number of synaptic failures and reduced the frequency of mIPSCs. The facilitation or suppression of GABAergic transmission by the GluR5 agonists did not require activation of voltage-gated calcium channels or presynaptic GABA(B) receptors. It was also found that extracellular, endogenous glutamate tonically reduces the rate of failures of GABAergic transmission. These results suggest that the terminals of GABAergic neurons in the BLA carry two subtypes of GluR5-containing kainate receptors, which have different agonist affinities and activate opposing mechanisms of action. The GluR5-mediated, bidirectional modulation of GABA release by glutamate in the BLA may play an important role in the regulation of synaptic plasticity and neuronal excitability in this structure, under normal and pathological conditions. PMID- 12533605 TI - BETA2/NeuroD1 null mice: a new model for transcription factor-dependent photoreceptor degeneration. AB - BETA2/NeuroD1 is a basic helix-loop-helix transcription factor that is expressed widely throughout the developing nervous system. Previous studies have shown that BETA2/NeuroD1 influences the fate of retinal cells in culture. To analyze the effect of BETA2/NeuroD1 on the structure and function of the retina, we examined a line of BETA2/NeuroD1 knock-out mice that survives until adulthood. At 2-3 months of age, homozygous null mice showed a 50% reduction in rod-driven electroretinograms (ERGs) and a 65% reduction in cone-driven ERGs. ERGs measured from knock-out mice that were >9 months of age were undetectable. At 2-3 months, the number of photoreceptors in the outer nuclear layer was reduced by 50%. In addition, electron microscopy showed that the surviving photoreceptors had shortened outer segments. The number of cones labeled by peanut agglutinin was decreased 50-60%. By 18 months, retinas from null mice were completely devoid of photoreceptors. There appeared to be few changes in the inner retina, although BETA2/NeuroD1 is expressed in this area. Terminal deoxynucleotidyl transferase mediated biotinylated UTP nick end labeling staining revealed a dramatic increase in cell death, peaking at approximately postnatal day 3 and continuing into adulthood. No defects in cell birth were detected using bromodeoxyuridine staining. Our results reveal that BETA2/NeuroD1 not only plays an important role in terminal differentiation of photoreceptors but also serves as a potential survival factor. Loss of BETA2/NeuroD1 results in an age-related degeneration of both rods and cones. PMID- 12533606 TI - In vivo and in vitro neurotoxicity of the human prion protein (PrP) fragment P118 135 independently of PrP expression. AB - We recently demonstrated that the 118-135 putative transmembrane domain of prion protein (PrP) exhibited membrane fusogenic properties and induced apoptotic neuronal cell death of rat cortical neurons, independently of its aggregation state. The aim of the present study was to analyze the in vivo neurotoxicity of the prion fragment P118-135 and to evaluate the potential role of the physiological isoform of PrP in the P118-135-induced cell death. Here, we demonstrate that the nonfibrillar P118-135 is cytotoxic to retinal neurons in vivo as monitored by intravitreal inoculation and recording of the electrical activity of retina and tissue examination. Moreover, knock-out PrP gene mice exhibit similar sensitivity to the nonfibrillar P118-135-induced cell death and electrical perturbations, strongly suggesting that cell death occurs independently of PrP expression. Interestingly, a variant nonfusogenic P118-135 peptide (termed P118-135theta) had no effects on in vivo neuronal viability, suggesting that the P118-135-induced cell death is mediated by its membrane destabilizing properties. These data have further been confirmed in vitro. We show that the fusogenic peptide P118-135 induces death of cultured neurons from both wild-type and knock-out PrP gene mice via an apoptotic-mediated pathway, involving early caspase activation and DNA fragmentation. Altogether these results emphasize the neurotoxicity of the fusogenic nonfibrillar PrP transmembrane domain and indicate that fibril formation and PrP expression are not obligatory requirements for neuronal cell death. The use of synthetic prion peptides could provide insights into the understanding of neuronal loss mechanisms that take place during the development of the various types of spongiform encephalopathies. PMID- 12533607 TI - Light stimulates a transducin-independent increase of cytoplasmic Ca2+ and suppression of current in cones from the zebrafish mutant nof. AB - Transducins couple visual pigments to cGMP hydrolysis, the only recognized phototransduction pathway in vertebrate photoreceptors. Here we describe a zebrafish mutant, no optokinetic response f(w21) (nof), with a nonsense mutation in the gene encoding the alpha subunit of cone transducin. Retinal morphology and levels of phototransduction enzymes are normal in nof retinas, but cone transducin is undetectable. Dark current in nof cones is also normal, but it is insensitive to moderate intensity light. The nof cones do respond, however, to bright light. These responses are produced by a light-stimulated, but transducin independent, release of Ca2+ into the cone cytoplasm. Thus, in addition to stimulating transducin, light also independently induces release of Ca2+ into the photoreceptor cytoplasm. PMID- 12533608 TI - Interleukin-6 protects anterior horn neurons from lethal virus-induced injury. AB - We evaluated the role of interleukin-6 (IL-6) in neuronal injury after CNS infection. IL-6-/- and IL-6+/+ mice of resistant major histocompatibility complex (MHC) H-2b haplotype intracerebrally infected with Theiler's virus cleared the infection normally without development of viral persistence, lethal neuronal infection, or late phase demyelination. In contrast, infection of IL-6-/- mice on a susceptible H-2q haplotype resulted in frequent deaths and severe neurologic deficits within 2 weeks of infection as compared with infected IL-6+/+ H-2q littermate controls. Morphologic analysis demonstrated dramatic injury to anterior horn neurons of IL-6-/- H-2q mice at 12 d after infection. Infectious viral titers in the CNS (brain and spinal cord combined) were equivalent between IL-6-/- H-2q and IL-6+/+ H-2q mice. In contrast, more viral RNA was detected in the spinal cord of IL-6-/- mice compared with IL-6+/+ H-2q mice. Virus antigen was localized predominantly to anterior horn cells in infected IL-6-/- H-2q mice. IL-6 deletion did not affect the humoral response directed against virus, nor did it affect the expression of CD4, CD8, MHC class I, or MHC class II in the CNS. Importantly, IL-6 was expressed by astrocytes of infected IL-6+/+ mice but not in astrocytes of IL-6-/- mice or uninfected IL-6+/+ mice. Furthermore, expression of various chemokines was robust at 12 d after infection in both H-2b and H-2q IL-6 /- mice, indicating that intrinsic CNS inflammatory responses did not depend on the presence of IL-6. Finally, in vitro analysis of virus-induced death in neuroblastoma-spinal cord-34 motor neurons and primary anterior horn cell neurons showed that IL-6 exerted a neuroprotective effect. These data support the hypothesis that IL-6 plays a critical role in protecting specific populations of neurons from irreversible injury. PMID- 12533609 TI - Aberrant activation of focal adhesion proteins mediates fibrillar amyloid beta induced neuronal dystrophy. AB - Neuronal dystrophy is a pathological hallmark of Alzheimer's disease (AD) that is not observed in other neurodegenerative disorders that lack amyloid deposition. Treatment of cortical neurons with fibrillar amyloid beta (Abeta) peptides induces progressive neuritic dystrophy accompanied by a marked loss of synaptophysin immunoreactivity (Grace et al., 2002). Here, we report that fibrillar Abeta-induced neuronal dystrophy is mediated by the activation of focal adhesion (FA) proteins and the formation of aberrant FA structures adjacent to Abeta deposits. In the AD brain, activated FA proteins are observed associated with the majority of senile plaques. Clustered integrin receptors and activated paxillin (phosphorylated at Tyr-31) and focal adhesion kinase (phosphorylated at Tyr-297) are mainly detected in dystrophic neurites surrounding Abeta plaque cores, where they colocalize with hyperphosphorylated tau. Deletion experiments demonstrated that the presence of the LIM domains in the paxillin C terminus and the recruitment of the protein-Tyr phosphatase (PTP)-PEST to the FA complex are required for Abeta-induced neuronal dystrophy. Therefore, both paxillin and PTP PEST appear to be critical elements in the generation of the dystrophic response. Paxillin is a scaffolding protein to which other FA proteins bind, leading to the formation of the FA contact and initiation of signaling cascades. PTP-PEST plays a key role in the dynamic regulation of focal adhesion contacts in response to extracellular cues. Thus, in the AD brain, fibrillar Abeta may induce neuronal dystrophy by triggering a maladaptive plastic response mediated by FA protein activation and tau hyperphosphorylation. PMID- 12533610 TI - Critical roles of thioredoxin in nerve growth factor-mediated signal transduction and neurite outgrowth in PC12 cells. AB - Thioredoxin (TRX) has a role in a variety of biological processes, including cytoprotection and the activation of transcription factors. Nerve growth factor (NGF) is a major survival factor of sympathetic neurons and promotes neurite outgrowth in rat pheochromocytoma PC12 cells. In this study, we showed that NGF induces TRX expression at protein and mRNA levels. NGF activated the TRX gene through a regulatory region positioned from -263 to -217 bp, containing the cAMP responsive element (CRE). Insertion of a mutation in the CRE in this region abolished the response to NGF. NGF induced binding of CRE-binding protein to the CRE of the TRX promoter in an electrophoretic mobility shift assay. NGF also induced nuclear translocation of TRX. 2'-Amino-3'-methoxyflavone, an inhibitor of mitogen-activated protein kinase kinase, which is a known inhibitor of NGF dependent differentiation in PC12 cells, suppressed the NGF-dependent expression and nuclear translocation of TRX. Overexpression of mutant TRX (32S/35S) or TRX antisense vector blocked the neurite outgrowth of PC12 cells by NGF. Overexpression of mutant TRX (C32S/C35S) suppressed the NGF-dependent activation of the CRE-mediated c-fos reporter gene. These results suggest that TRX plays a critical regulatory role in NGF-mediated signal transduction and outgrowth in PC12 cells. PMID- 12533611 TI - Correlation between brain reorganization, ischemic damage, and neurologic status after transient focal cerebral ischemia in rats: a functional magnetic resonance imaging study. AB - The pattern and role of brain plasticity in stroke recovery has been incompletely characterized. Both ipsilesional and contralesional changes have been described, but it remains unclear how these relate to functional recovery. Our goal was to correlate brain activation patterns with tissue damage, hemodynamics, and neurologic status after temporary stroke, using functional magnetic resonance imaging (fMRI). Transverse relaxation time (T2)-weighted, diffusion-weighted, and perfusion MRI were performed at days 1 (n = 7), 3 (n = 7), and 14 (n = 7) after 2 hr unilateral middle cerebral artery occlusion in rats. Functional activation and cerebrovascular reactivity maps were generated from contrast-enhanced fMRI during forelimb stimulation and hypercapnia, respectively. Before MRI, rats were examined neurologically. We detected loss of activation responses in the ipsilesional sensorimotor cortex, which was related to T2 lesion size (r = -0.858 on day 3, r = -0.979 on day 14; p < 0.05). Significant activation responses in the contralesional hemisphere were detected at days 1 and 3. The degree of shift in balance of activation between the ipsilesional and contralesional hemispheres, characterized by the laterality index, was linked to the T2 and apparent diffusion coefficient in the ipsilesional contralesional forelimb region of the primary somatosensory cortex and primary motor cortex at day 1 (r = -0.807 and 0.782, respectively; p < 0.05) and day 14 (r = -0.898 and -0.970, respectively; p < 0.05). There was no correlation between activation parameters and perfusion status or cerebrovascular reactivity. Finally, we found that the laterality index and neurologic status changed in parallel over time after stroke, so that when all time points were grouped together, neurologic status was inversely correlated with the laterality index (r = -0.571; p = 0.016). This study suggests that the degree of shift of activation balance toward the contralesional hemisphere early after stroke increases with the extent of tissue injury and that functional recovery is associated mainly with preservation or restoration of activation in the ipsilesional hemisphere. PMID- 12533612 TI - Vesicular neurotransmitter transporter expression in developing postnatal rodent retina: GABA and glycine precede glutamate. AB - Vesicular transporters regulate the amount and type of neurotransmitter sequestered into synaptic vesicles and, hence, the kind of signal transmitted to postsynaptic neurons. Glutamate is the prominent excitatory neurotransmitter in retina; GABA and glycine are the main inhibitory neurotransmitters. Little is known about the ontogeny of vesicular neurotransmission in retina. We investigated expression of glutamatergic [vesicular glutamate transporter 1 (VGLUT1)] and GABA/glycinergic [vesicular GABA/glycine transporter (VGAT)] vesicular transporters in postnatal retina. VGLUT1 labels glutamatergic synapses. VGLUT1 and synaptic vesicle 2 colocalized to photoreceptor terminals. VGLUT1 colocalized with PKC to rod bipolar terminals and to ON bipolar terminals in metabotropic glutamate receptor 6+/- mice. Developmentally, VGAT expression precedes VGLUT1. In rat and mouse retina, VGAT occurred in the inner retina by postnatal day 1 (P1). In rat retina, VGLUT1 was in the outer retina by P5-P7 and the inner retina by P7. In the mouse retina, VGLUT1 expression was in the outer retina by P3 and the inner retina by P5. Both rat and mouse retina had an adult pattern of VGLUT1 expression by P14. VGLUT1 expression precedes ribbon synapses, which are first observed in the inner retina at P11 (Fisher, 1979) in mouse and P13 (Horsburgh and Sefton, 1987) in rat. The ribbon synapse marker RIBEYE was not detected in inner retina of P5 or P7 rat. Spontaneous EPSCs in mouse ganglion cells were recorded as early as P7. Together, these findings indicate that vesicular GABA and glycine transmission precedes vesicular glutamate transmission in developing rodent retina. Furthermore, vesicular glutamate transmission likely occurs before ribbon synapse formation in the inner retina. PMID- 12533613 TI - The L1 cell adhesion molecule is essential for topographic mapping of retinal axons. AB - The retinocollicular projection is a preferred axon guidance pathway for investigating molecular mechanisms of synaptic targeting in the mammalian CNS. Here we identify a previously unrecognized role of the L1 cell adhesion molecule in topographic mapping of retinal ganglion cell (RGC) axons to their targets in the mouse superior colliculus (SC). L1 was transiently expressed on RGC axons during axon growth and targeting. DiI labeling of retinal axons revealed that temporal axons of L1-minus mice bypassed correct target locations in the anterior SC, forming termination zones at incorrect posterior sites, which were often skewed along the mediolateral axis. During development of the retinotopic map L1 minus temporal axons extended across the anteroposterior axis of the SC like wild type axons but failed to arborize at normal anterior target sites. L1-minus RGC axons exhibited normal crossing at the optic chiasm and fasciculation of the optic nerve. Results suggest that retinal axons require the function of L1 in addition to repellent EphA guidance receptors to achieve proper topographic mapping. PMID- 12533614 TI - Aberrant patterning of neuromuscular synapses in choline acetyltransferase deficient mice. AB - In this study we examined the developmental roles of acetylcholine (ACh) by establishing and analyzing mice lacking choline acetyltransferase (ChAT), the biosynthetic enzyme for ACh. As predicted, ChAT-deficient embryos lack both spontaneous and nerve-evoked postsynaptic potentials in muscle and die at birth. In mutant embryos, abnormally increased nerve branching occurs on contact with muscle, and hyperinnervation continues throughout subsequent prenatal development. Postsynaptically, ACh receptor clusters are markedly increased in number and occupy a broader muscle territory in the mutants. Concomitantly, the mutants have significantly more motor neurons than normal. At an ultrastructural level, nerve terminals are smaller in mutant neuromuscular junctions, and they make fewer synaptic contacts to the postsynaptic muscle membrane, although all of the typical synaptic components are present in the mutant. These results indicate that ChAT is uniquely essential for the patterning and formation of mammalian neuromuscular synapses. PMID- 12533615 TI - Neutral amino acid transporter ASCT1 is preferentially expressed in L-Ser synthetic/storing glial cells in the mouse brain with transient expression in developing capillaries. AB - Nonessential amino acid L-Ser plays an essential role in neuronal survival and differentiation, through preferential expression of the L-Ser biosynthetic enzyme 3-phosphoglycerate dehydrogenase (3PGDH), in particular in glial cells but not in neurons. To seek the molecular candidates responsible for glia-borne L-Ser transport, we performed histochemical analyses on amino acid transporter ASCT1, which prefers small neutral amino acids, such as Ala, Ser, Cys, and Thr, and mediates their obligatory exchange. At early developmental stages, neuroepithelial cells constituting the ventricular zone expressed ASCT1 mRNA and protein ubiquitously. Thereafter, ASCT1 expression was gradually downregulated in neuronal populations during the late embryonic and neonatal periods, whereas its high expression was transmitted to radial glial cells and then to astrocytes. High levels of ASCT1 were also detected in the olfactory ensheathing glia. The preferential glial expression of ASCT1 was consistent with that of 3PGDH, and their extensive colocalization was demonstrated at the cellular level. Moreover, high cellular contents of L-Ser were revealed in these glial cells by using a specific antibody to L-Ser. These results strongly suggest that a large amount of L-Ser is synthesized and stored in these glial cells and is released through ASCT1 in exchange for other extracellular substrates. In addition, we observed prominent expression of ASCT1 in capillary endothelial cells of embryonic and neonatal brains. Therefore, ASCT1 appears to be regulated to meet metabolic demands by differentiating and mature neurons through the transport of glia- and blood-borne small neutral amino acids. PMID- 12533616 TI - Glial cell line-derived neurotrophic factor alters axon schwann cell units and promotes myelination in unmyelinated nerve fibers. AB - Glial cell line-derived neurotrophic factor (GDNF) plays an important role in the development and maintenance of a subset of dorsal root ganglion sensory neurons. We administered high-dose exogenous recombinant human GDNF (rhGDNF) daily to adult rats to examine its effect on unmyelinated axon-Schwann cell units in intact peripheral nerves. In rhGDNF-treated animals, there was a dramatic proliferation in the Schwann cells of unmyelinated fibers, which resulted in the segregation of many unmyelinated axons into a 1:1 relationship with Schwann cells and myelination of normally unmyelinated small axons. This study demonstrates that the administration of high doses of a growth factor to adult rats can change the phenotype of nerve fibers from unmyelinated to myelinated. PMID- 12533617 TI - DLX5 regulates development of peripheral and central components of the olfactory system. AB - Induction, neurogenesis, and synaptogenesis of the olfactory bulb are thought to require interactions with the olfactory epithelium. The Dlx family of homeobox genes is expressed in both the olfactory bulb and olfactory epithelium. In particular, Dlx5 is expressed in the olfactory placode, olfactory epithelium, and local circuit neurons of the olfactory bulb. Here we analyzed mice lacking DLX5 function. The Dlx5-/- mutation reduces the size of the olfactory epithelium. Although some olfactory neurons are formed, they fail to generate olfactory axons that innervate the olfactory bulb. Despite the lack of innervation, the olfactory bulb forms, and neurogenesis of projection and local circuit neurons proceeds. However, the mutation has a cell-autonomous effect on the ability of neural progenitors to produce olfactory bulb local circuit neurons, with granule cells more severely affected than periglomerular cells. In addition, the mutation has a noncell-autonomous effect on the morphogenesis of mitral cells. PMID- 12533618 TI - A novel skilled-reaching impairment in paw supination on the "good" side of the hemi-Parkinson rat improved with rehabilitation. AB - Parkinson's disease is characterized by tremor, rigidity, bradykinesia, and postural abnormalities ascribed to the loss of nigrostriatal dopamine (DA). Symptoms similar to the human condition can be produced in the rat by DA depleting 6-hydroxydopamine injections made into the nigrostriatal system. After a unilateral lesion, the rat symptoms include sensory and motor impairments and turning biases reflecting motor abnormalities to the contralateral-to-depletion side of the body. In addition, a number of studies on skilled reaching report impairments in the use of the ipsilateral limb. It is suggested that the ipsilateral deficit is secondary to the contralateral motor impairments however. Here we re-examine how rats with unilateral DA depletion use their ipsilateral limb for skilled reaching for food. We provide the first description of an impairment on the ipsilateral-to-depletion side of the body of the rat and the first demonstration of amelioration of the defect using behavioral therapy. Video analysis of rats reaching for single pellets of food with the ipsilateral limb revealed that, although limb advancement and food grasping were normal, paw supination and food release to the mouth were impaired. Consequently, the animals were unable to transport a grasped food pellet to the mouth. Behavioral therapy, consisting of training in a simpler reaching task, strikingly lessened the impairment and improved reaching movements to the point that the rats could transport the food to the mouth. The results are discussed in relation to possible causes of the ipsilateral impairment, its treatment, and to relevant research on human Parkinson patients, indicating that they display bilateral improvements after unilateral treatments. PMID- 12533619 TI - Characterization of the circuits that generate spontaneous episodes of activity in the early embryonic mouse spinal cord. AB - In the developing nervous system, patterned spontaneous activity affects a variety of developmental processes. Thus, it is important to identify the earliest time that such activity occurs and to characterize the underlying circuitry. In isolated mouse spinal cord-limb preparations, highly rhythmic spontaneous activity occurred as early as embryonic day 11 (E11)-E12, when many lumbosacral motoneurons were still migrating and extending their peripheral projections. This activity required both electrical and chemical transmission, and acetylcholine, rather than glutamate, provided the main excitatory drive. Our data are consistent with motoneurons themselves playing a critical role in generating such activity by making excitatory connections on each other and on GABAergic interneurons via dihydro-beta-erythroidine hydrobromide (DHbetaE) insensitive nicotinic receptors. This resulted in the generation of local bursts. Consistent with these observations, E12-E12.5 mouse motoneurons retrogradely labeled by HRP were observed to have extensive axon collaterals that projected locally within the lateral motor column and to interneuron-containing regions dorsal and medial of the lateral motor column. Cholinergic axons, presumably from motoneurons, were also observed in the ventral and lateral funiculi. However, for local bursts to propagate throughout the cord, a second DHbetaE-sensitive cholinergic pathway that also involved glycinergic interneurons was required. This circuit characterization should facilitate the use of genetic mutations that alter specific subpopulations of interneurons or cholinergic transmission to determine how modifying different aspects of this early activity affects subsequent development of the spinal motor circuit. PMID- 12533620 TI - Widespread projections from myelinated nociceptors throughout the substantia gelatinosa provide novel insights into neonatal hypersensitivity. AB - Skin sensory neurons have long been thought to undergo major changes in anatomy and physiology over the first few weeks of postnatal life. Low-threshold mechanoreceptors (LTMRs) are believed to project extensively throughout superficial dorsal horn laminas initially and provide the afferent limb for hyperactive nocifensive reflexes. However, our recent studies revealed that neonatal LTMRs do not project into "pain-specific" regions; instead, they exhibit adult-like anatomy shortly after birth. We sought to determine whether the same might be true for myelinated high-threshold mechanoreceptors (HTMRs). We used an intact, ex vivo somatosensory system preparation from neonatal mice to allow intrasomal recording and neurobiotin labeling of individual sensory neurons characterized via natural skin stimuli. Neonatal HTMRs displayed a number of key hallmarks of their adult counterparts; relative to LTMRs, they exhibited broader, inflected somal spikes and higher mechanical thresholds and/or responded in an increasingly vigorous manner to incrementally graded forces in a manner capable of encoding stimulus intensity. Two types were discerned on the basis of central anatomy: one subset projected to superficial laminas (I/II); the other gave rise to diffuse, dorsally recurving collateral arbors extending throughout the entire dorsal horn (I-V). The latter represent a novel cutaneous afferent morphology that persists in older animals. These studies reveal that inputs from myelinated afferents to superficial pain-specific laminas in neonates arise from HTMRs and not LTMRs as commonly thought. This frequently overlooked population is in a position, therefore, to contribute substantially to paradoxical nocifensive behaviors in neonates and various pain states in adults. PMID- 12533621 TI - Muscarinic blockade slows and degrades the location-specific firing of hippocampal pyramidal cells. AB - The firing of rat hippocampal pyramidal cells is determined both by the animal's location and by the state of the hippocampal EEG. Because cholinergic transmission plays a role in EEG activity, we expected that its modification would alter place cell activity. We therefore investigated the effects on place cell activity of blocking muscarinic transmission with intracerebroventricular injections of scopolamine. Scopolamine reduced both the rate of place cell discharge inside firing fields and the spatial coherence of the fields; discharge outside of the fields also showed small increases. After injections, fields were shifted farther from their previous location than for saline controls, indicating reduced reproducibility after muscarinic blockade. Scopolamine increased the time rats were stationary, but changes in place cell activity persisted even after analysis was restricted to periods of walking, suggesting that the behavioral changes cannot account for the cell discharge changes. The scopolamine effects were dose dependent to an extent that varied between different measures. The firing rates of interneurons showed only a minor trend to decrease after scopolamine. Nevertheless, the spatial coherence of interneuron firing patterns was reduced, consistent with the recent demonstration that their positional firing is mediated by the location-specific firing of pyramids (Marshall et al., 2002). These results demonstrate that acetylcholine enhances positional firing patterns in the hippocampus. Muscarinic blockade weakens the positional firing of most place cells and therefore renders them less useful for precise representation of the environment. This effect may underlie the difficulties in spatial learning and problem solving caused by abnormalities of cholinergic transmission. PMID- 12533622 TI - Genetic disruption of cortical interneuron development causes region- and GABA cell type-specific deficits, epilepsy, and behavioral dysfunction. AB - The generation of properly functioning circuits during brain development requires precise timing of cell migration and differentiation. Disruptions in the developmental plan may lead to neurological and psychiatric disorders. Neocortical circuits rely on inhibitory GABAergic interneurons, the majority of which migrate from subcortical sources. We have shown that the pleiotropic molecule hepatocyte growth factor/scatter factor (HGF/SF) mediates interneuron migration. Mice with a targeted mutation of the gene encoding urokinase plasminogen activator receptor (uPAR), a key component in HGF/SF activation and function, have decreased levels of HGF/SF and a 50% reduction in neocortical GABAergic interneurons at embryonic and perinatal ages. Disruption of interneuron development leads to early lethality in most models. Thus, the long-term consequences of such perturbations are unknown. Mice of the uPAR-/- strain survive until adulthood, and behavior testing demonstrates that they have an increased anxiety state. The uPAR-/- strain also exhibits spontaneous seizure activity and higher susceptibility to pharmacologically induced convulsions. The neocortex of the adult uPAR-/- mouse exhibits a dramatic region- and subtype specific decrease in GABA-immunoreactive interneurons. Anterior cingulate and parietal cortical areas contain 50% fewer GABAergic interneurons compared with wild-type littermates. However, interneuron numbers in piriform and visual cortical areas do not differ from those of normal mice. Characterization of interneuron subpopulations reveals a near complete loss of the parvalbumin subtype, with other subclasses remaining intact. These data demonstrate that a single gene mutation can selectively alter the development of cortical interneurons in a region- and cell subtype-specific manner, with deficits leading to long-lasting changes in circuit organization and behavior. PMID- 12533623 TI - The influence of behavioral context on the representation of a perceptual decision in developing oculomotor commands. AB - To make decisions about sensory stimuli, the brain must weigh the evidence that supports or opposes the alternative interpretations. In the present study, we evaluated the hypothesis that a quantity reflecting the weight of sensory evidence is represented in brain circuits responsible for the behavioral response used to indicate the decision. We trained monkeys to decide the direction of random-dot motion and to indicate their decision with an eye movement to one of two choice targets. We interrupted decision formation with electrical microstimulation of the frontal eye field, causing an evoked eye movement that is influenced by ongoing oculomotor activity. For the "pro-saccade" version of the task, in which the correct target was at a known location in the direction of motion, the microstimulus-evoked eye movement reflected both the impending pro saccadic response and the temporal accumulation of motion information used to select that response. In contrast, for the "colored-target" task, in which the correct target was of a particular color but at an unpredictable location, little ongoing oculomotor activity was evident. The results suggest that formation of the decision and formation of the behavioral response share a common level of neural organization, but only when the decision is associated with a specific, predictable movement. PMID- 12533624 TI - Intrinsic role of polysialylated neural cell adhesion molecule in photic phase resetting of the Mammalian circadian clock. AB - The suprachiasmatic nuclei (SCN), the location of the mammalian circadian clock, are one of the few adult brain regions that express the highly polysialylated form of neural cell adhesion molecule (PSA-NCAM). A role for the polysialic acid (PSA) component of PSA-NCAM, which is known to promote tissue plasticity, has been reported for photic entrainment of circadian rhythmicity in vivo. The in vivo results, however, do not discriminate between PSA acting upstream or downstream of the glutamatergic synapses that convey photic information to the SCN. To address this key issue, we exploited an in vitro rat brain slice preparation that retains robust circadian function. As in the intact SCN, PSA levels in the isolated SCN are rhythmic, with higher levels during the early subjective day and lower levels during subjective night. Importantly, bath application of glutamate to SCN slices rapidly and transiently increases PSA levels during both the subjective day and night. Pretreating the slices with endoneuraminidase, which selectively removes PSA from NCAM and thereby prevents this increase, abolishes glutamate- and optic chiasm stimulation-induced phase delays of the SCN circadian neuronal activity rhythm. These results support the hypothesis that PSA expression in the SCN is controlled by both the circadian clock and photic input to the clock and that expression of PSA in the SCN is critical for photic-like phase shifts of the clock. Together, these results establish that such actions of PSA are manifested downstream from presynaptic retinohypothalamic terminals and therefore are intrinsic to the SCN itself. PMID- 12533625 TI - Associative memory formation increases the observation of dendritic spines in the hippocampus. AB - Dendritic spines are sources of synaptic contact that can be altered by experience and, as such, may be involved in memories for that experience. Here we tested whether the acquisition of new memories is associated with changes in the density of dendritic spines. Adult male rats were trained using the trace eyeblink conditioning paradigm, an associative learning task that requires the hippocampus for acquisition. Additional groups were exposed to the same number of stimuli presented in an explicitly unpaired manner or were naive. Twenty-four hours later, the density of dendritic spines was measured using Golgi impregnation. Trace conditioning was associated with an increase in the density of dendritic spines on the pyramidal cells of area CA1 of the hippocampus, an effect that was prevented by blocking acquisition of the learned response with a competitive NMDA receptor antagonist. Training with delay conditioning, a similar task that does not require the hippocampus, also produced an increase in spine density. The learning-induced increase in dendritic spine density was specific to basal dendrites of pyramidal cells in the CA1 region of the hippocampus. Changes did not occur on their apical dendrites or on cells in the dentate gyrus or somatosensory cortex. These results suggest that the formation and expression of associative memories increase the availability of dendritic spines and the potential for synaptic contact. PMID- 12533626 TI - The effect of lesions of the basolateral amygdala on instrumental conditioning. AB - In three experiments, we assessed the effect of lesions of the amygdala basolateral complex (BLA) on instrumental conditioning in rats. In experiment 1, the lesion had no effect on the acquisition of either lever pressing or chain pulling in food-deprived rats whether these actions earned food pellets or a maltodextrin solution. The lesion did attenuate, however, the impact of outcome devaluation, induced by sensory-specific satiety, on instrumental performance both when assessed in extinction and when reward was delivered contingent on instrumental performance. In experiment 2, evidence was found to suggest that the lesioned rats differed from shams in their ability to encode the specific action outcome contingencies to which they were exposed during training: lesioned rats failed to adjust their performance appropriately when the action-outcome contingency was degraded. These effects were not caused by an inability of BLA lesioned rats to discriminate the two instrumental actions; these rats were similar to shams in their acquisition of a heterogeneous instrumental chain involving lever pressing and chain pulling (experiment 3). In experiment 4, however, lesions of the BLA were found to produce a deficit in the ability of rats to use the specific properties of the instrumental outcomes used in the previous experiments to discriminate rewarded from unrewarded actions in a free operant discrimination situation. Together these results suggest that in instrumental conditioning, the BLA mediates outcome encoding, specifically relating the sensory features of nutritive commodities to the emotional consequences induced by their consumption. PMID- 12533627 TI - Perceptual attentional set-shifting is impaired in rats with neurotoxic lesions of posterior parietal cortex. AB - The posterior parietal cortex (PPC) is believed to be involved in the representation of spatial information, including spatial attentional processing. Because the PPC is extensively interconnected with frontal cortical regions involved in attention and executive function, we sought to determine whether PPC was involved in nonspatial attentional processes such as those of the frontal areas to which it projects. Lesions of the medial frontal cortex (in rats) or lateral prefrontal cortex (in nonhuman primates) impair the ability to shift attention from one perceptual dimension of a stimulus to another (referred to as an extradimensional shift). Rats with neurotoxic lesions of the PPC tested in an attentional set-shifting paradigm demonstrated a pattern of impairment identical to that of rats with medial frontal cortex lesions: they were selectively impaired on the extradimensional shift phase of the task. Performance in other phases of the task was indistinguishable from that of control rats, including the ability to reverse a previously learned discrimination. These findings are consistent with models that assign the PPC a prominent role in cortical attentional processing networks, as well as a role for the PPC in processing information about expectancy and surprise. They also suggest, importantly, that the interaction between the PPC and the frontal cortex is not limited to spatial attentional processing. PMID- 12533628 TI - Coactivation of motoneurons regulated by a network combining electrical and chemical synapses. AB - Electrical transmission among neurons has been considered a mechanism to synchronize neuronal activity, and rectification provides a mechanism to confine the flow of signals among the connected neurons. The question is how this type of transmission operates within complex neuronal networks. In the leech, the neurons located in position 151 of the midbody ganglion map are connected to virtually every motoneuron via rectifying electrical synapses that pass negative current to the motoneurons. These are nonspiking neurons, and here we have labeled them NS neurons. The goal of this investigation has been to assess their role in regulating motor activity and how rectifying electrical synapses contribute to the function of motor networks. The coupling between NS neurons and motoneurons was voltage sensitive: it increased as motoneurons were depolarized. In addition, excitation of motoneurons evoked hyperpolarizing synaptic responses in NS neurons, the amplitude of which depended on the membrane potential of the latter and on the motoneuron firing frequency. This hyperpolarization was mediated by chemical transmission through an interneuronal layer that spanned the nerve cord. These interactions established a feedback loop between NS and motoneurons that was regulated by the membrane potential of NS. This mechanism was responsible for the uncoupling between otherwise electrically coupled motoneurons. In this way, the NS neurons can act as "electrical neuromodulators," modifying the interaction of other neurons, depending on the activity of the system as a whole. PMID- 12533629 TI - Effects of sex and estrogen on behavioral sensitization to cocaine in rats. AB - Estrogen rapidly enhances dopamine (DA) activity in the striatum and nucleus accumbens as well as behavioral responses to psychomotor stimulants in female rats but not males. This experiment was conducted to investigate the role of pulsatile estrogen treatment on and sex differences in the development and expression of sensitization of cocaine-induced rotational behavior in rats with unilateral striatal DA denervation. Four groups were tested: ovariectomized (OVX) females treated with 5 microg of estradiol benzoate (OVX+E), OVX females, castrated (CAST) males, and intact males. Animals received estrogen or vehicle 30 min before cocaine (0, 5, 10, or 20 mg/kg, i.p.) on 4 consecutive days, followed by 3 d without treatment for 3 weeks. At the conclusion of the experiment, animals were withdrawn from hormone and/or cocaine for 10 d, and all groups underwent a challenge test with 10 mg/kg cocaine. We report here that OVX+E females exhibit significantly greater sensitization of rotational behavior with a faster rate of sensitization than the three other groups. There is also a sex difference independent of gonadal hormones: OVX females exhibit a greater magnitude of sensitization of rotational behavior than do CAST males at 20 mg/kg cocaine. Furthermore, on the challenge test, OVX+E animals tested without estrogen treatment continue to exhibit greater rotational behavior than do all other groups. Thus, estrogen enhances sensitization to cocaine, there are sex differences in behavioral sensitization, and sensitization that develops under conditions with estrogen persists even when estrogen levels are low. PMID- 12533630 TI - Corticotropin-releasing factor receptors couple to multiple G-proteins to activate diverse intracellular signaling pathways in mouse hippocampus: role in neuronal excitability and associative learning. AB - Corticotropin-releasing factor (CRF) exerts a key neuroregulatory control on stress responses in various regions of the mammalian brain, including the hippocampus. Using hippocampal slices, extracts, and whole animals, we investigated the effects of human/rat CRF (h/rCRF) on hippocampal neuronal excitability and hippocampus-dependent learning in two mouse inbred strains, BALB/c and C57BL/6N. Intracellular recordings from slices revealed that application of h/rCRF increased the neuronal activity in both mouse inbred strains. Inhibition of protein kinase C (PKC) by bisindolylmaleimide I (BIS-I) prevented the h/rCRF effect only in hippocampal slices from BALB/c mice but not in slices from C57BL/6N mice. Inhibition of cAMP-dependent protein kinase (PKA) by H-89 abolished the h/rCRF effect in slices from C57BL/6N mice, with no effect in slices from BALB/c mice. Accordingly, h/rCRF elevated PKA activity in hippocampal slices from C57BL/6N mice but increased only PKC activity in the hippocampus of BALB/c mice. These differences in h/rCRF signal transduction were also observed in hippocampal membrane suspensions from both mouse strains. In BALB/c mice, hippocampal CRF receptors coupled to G(q/11) during stimulation by h/rCRF, whereas they coupled to G(s), G(q/11), and G(i) in C57BL/6N mice. As expected on the basis of the slice experiments, h/rCRF improved context-dependent fear conditioning of BALB/c mice in behavioral experiments, and BIS-I prevented this effect. However, although h/rCRF increased neuronal spiking in slices from C57BL/6N mice, it did not enhance conditioned fear. These results indicate that the CRF system activates different intracellular signaling pathways in mouse hippocampus and may have distinct effects on associative learning depending on the mouse strain investigated. PMID- 12533631 TI - Absence of thermal hyperalgesia in serotonin transporter-deficient mice. AB - Antidepressants in the treatment of neuropathic pain are thought to partially exert their effect by inhibition of serotonin (5-HT) reuptake and thus activation of central antinociceptive pathways. Mice deficient for the 5-HT transporter (5 HTT-/- mice) are regarded as a model of lifelong treatment with a serotonin reuptake inhibitor. Here we investigated 5-HTT-/- mice and compared their pain related behavior after a unilateral chronic constrictive sciatic nerve injury (CCI) with that of wild-type littermates. Wild-type mice reproducibly developed ipsilateral thermal hyperalgesia and mechanical allodynia after CCI. 5-HTT-/- mice did not develop thermal hyperalgesia, but showed bilateral mechanical allodynia after the nerve injury. 5-HT levels as measured with HPLC increased after CCI in the injured nerve in both genotypes and decreased in the lumbar spinal cord in wild-type mice. 5-HTT-/- mice had significantly lower 5-HT concentrations than wild-type mice in all tissues investigated. Thus, in 5-HTT-/- mice, reduced 5-HT levels in the injured peripheral nerves correlate with diminished behavioral signs of thermal hyperalgesia, a pain-related symptom caused by peripheral sensitization. In contrast, bilateral mechanical allodynia, a centrally mediated phenomenon, was associated with decreased spinal 5-HT concentrations in 5-HTT-/- mice and may possibly be caused by a lack of spinal inhibition. PMID- 12533632 TI - Interaural time difference discrimination thresholds for single neurons in the inferior colliculus of Guinea pigs. AB - Sensitivity to changes in the interaural time difference (ITD) of 50 msec tones was measured in single units in the inferior colliculus of urethane-anesthetized guinea pigs. ITD functions were measured with 100 repeats and fine spacing (100 points per cycle). The just noticeable difference (jnd) for ITD was determined using receiver operating characteristic (ROC) analysis of the spike-count distribution at each ITD. The jnd became progressively smaller as the signal frequency increased from 50 to 800 Hz but became unmeasurable above 1 kHz. The lowest jnds (30 microsec) were comparable with human jnds, indicating that there is sufficient information in the firings of individual neurons to permit discrimination without obligatory pooling. ROC analysis requires the choice of a reference ITD from which the jnd may be found by stepping the target ITD through the ITD function. For each neuron the reference was chosen to minimize the jnd. The lowest jnd was usually for ipsilateral leading references, near the minimum of the ITD function where the variance was also low, but where the slope was nearing its steepest. This was despite the peak of the ITD function occurring for contralateral leading stimuli. When the reference ITD was on midline, a jnd could be obtained by looking for firing rates either greater or smaller than the firing rate at midline. The lower jnd was usually obtained by looking for a decrease in firing rate. As duration increased, jnds either decreased or increased, depending on unit type, whereas when level increased, jnds generally increased. PMID- 12533636 TI - Peripheral T-cell lymphoma: spectrum of imaging findings with clinical and pathologic features. AB - Most radiologists are unfamiliar with peripheral T-cell lymphoma (PTCL) because PTCL represents a relatively small proportion of lymphomas and has a lower prevalence in Western countries. The World Health Organization classification of lymphoid neoplasms announced in 1999 resolved criticisms about lymphoma classification and aroused new interest in PTCL. The specific clinicopathologic entities of PTCL have particular primary locations and particular clinical and pathologic features. Radiologic images of patients with pathologically proved PTCL were retrospectively reviewed; clinical and pathologic data were also reviewed. PTCL involves various organs including the sinonasal cavity, airway, intestinal tract, skin, lymph nodes, liver, lung, and musculoskeletal system. The pattern of disease involvement in PTCL is not random. There is a correlation between specific clinicopathologic entities and the primary site of involvement, although the findings in the disseminated stage of disease do not allow differential diagnosis. It is significant that the radiologic features or locations of several entities are different from those of lymphoma with the B cell phenotype. Radiologic demonstration of disease progression beyond the primary site is clinically important because systemic dissemination in most of the entities leads to a dramatic change in the prognosis. PMID- 12533638 TI - Neurogenic tumors in the abdomen: tumor types and imaging characteristics. AB - There is a broad spectrum of neurogenic tumors that involve the abdomen. These tumors can be classified as those of (a) ganglion cell origin (ganglioneuromas, ganglioneuroblastomas, neuroblastomas), (b) paraganglionic system origin (pheochromocytomas, paragangliomas), and (c) nerve sheath origin (neurilemmomas, neurofibromas, neurofibromatosis, malignant nerve sheath tumors). Abdominal neurogenic tumors are most commonly located in the retroperitoneum, especially in the paraspinal areas and adrenal glands. All of these tumors except neuroblastomas and ganglioneuroblastomas are seen in adult patients. Abdominal neurogenic tumor commonly manifests radiologically as a well-defined, smooth or lobulated mass. Calcification may be seen in all types of neurogenic tumors. The diagnosis of abdominal neurogenic tumor is suggested by the imaging appearance of the lesion, including its location, shape, and internal architecture. Benign and malignant neurogenic tumors are difficult to differentiate unless distant metastatic foci are seen. For malignant tumors, imaging modalities other than computed tomography (CT) and magnetic resonance (MR) imaging may be necessary for staging. However, because most neurogenic tumors in adults are benign, CT and MR imaging can be used to develop a differential diagnosis and help determine the immediate local extent of tumor. PMID- 12533639 TI - Primary retroperitoneal neoplasms: CT and MR imaging findings with anatomic and pathologic diagnostic clues. AB - Primary retroperitoneal neoplasms are a rare but diverse group of benign and malignant tumors that arise within the retroperitoneal space but outside the major organs in this space. Although computed tomography and magnetic resonance imaging can demonstrate important characteristics of these tumors, diagnosis is often challenging for radiologists. Diagnostic challenges include precise localization of the lesion, determination of the extent of invasion, and characterization of the specific pathologic type. The first step is to determine whether the tumor is located within the retroperitoneal space. Displacement of normal anatomic structures of the retroperitoneum is helpful in this regard. For tumors that are located within the retroperitoneum, the next step is to identify the organ of origin. Specific signs, including the "beak sign," the "embedded organ sign," and the "phantom (invisible) organ sign," are useful for this purpose. When there is no definite sign that suggests the organ of origin, the diagnosis of a primary retroperitoneal tumor becomes likely. Awareness of specific patterns of spread, specific tumor components, and tumor vascularity help in further narrowing the differential diagnosis. Attention to these diagnostic clues is essential in making an accurate radiologic diagnosis of primary retroperitoneal tumors and in obtaining clinically significant information. PMID- 12533641 TI - US of gastrointestinal tract abnormalities with CT correlation. AB - Ultrasonography (US) is often the first imaging study performed in patients with abdominal pain or vague symptoms related to the gastrointestinal tract. An awareness of the US appearances of diseases of the intestine is essential to achieve the proper diagnosis and to enable appropriate triage of cases. Pathologic processes that affect the intestine generally result in decreased peristalsis and bowel wall thickening, both of which tend to decrease the luminal gas content. These changes permit evaluation of the intestine and surrounding structures with transabdominal and transvaginal US. US is useful in diagnosis of infectious and inflammatory conditions, such as appendicitis, Crohn disease, diverticulitis, epiploic appendagitis, pseudomembranous colitis, small bowel obstruction, small bowel vasculitis, and celiac disease. US is also helpful in diagnosis of tumors, such as gastric cancer, bowel lymphoma, and colon cancer. Familiarity with the US appearances of diseases that affect the intestine may allow specific diagnosis based on the degree and distribution of bowel wall thickening and associated changes of the perienteric tissues. PMID- 12533643 TI - Current role of CT in imaging of the stomach. AB - Recent advances in computed tomographic (CT) technology and three-dimensional (3D) imaging software have sparked renewed interest in using CT to evaluate gastric disease. Multidetector row CT scanners allow thinner collimation, which improves the visualization of subtle tumors as well as the quality of the 3D data sets. When water is used as an oral contrast agent, subtle disease is easier to visualize, especially when a rapid contrast material bolus is intravenously administered. Adenocarcinoma is the most common gastric malignancy and typically appears as focal or segmental wall thickening or a discrete mass. Gastric lymphoma can have a CT appearance similar to that of adenocarcinoma. Both gastric adenocarcinoma and lymphoma may be associated with adenopathy. Gastrointestinal stromal tumors (GISTs) tend to appear as well-defined masses that arise from the gastric wall and may be exophytic when large. GISTs are usually not associated with significant adenopathy. In addition to gastric malignancies, CT can also help detect inflammatory conditions of the stomach, including gastritis and peptic ulcer disease. CT angiography is especially helpful for depicting the gastric vasculature, which may be affected by a variety of disease conditions. PMID- 12533645 TI - Balloon dilation and stent placement for esophageal lesions: indications, methods, and results. AB - Esophageal balloon dilation and expandable stent placement are safe, minimally invasive, effective treatments for esophageal strictures and fistulas. These procedures have brought the management of dysphagia due to esophageal strictures into the field of interventional radiology. Esophageal dilation is usually indicated for benign stenoses and is technically successful in more than 90% of cases. Most patients with esophageal carcinoma are not candidates for resection; thus, the main focus of treatment is palliation of malignant dysphagia and esophagorespiratory fistulas. Esophageal stent placement, which is approved only for malignant strictures, is one of the main therapeutic options in affected patients and relieves dysphagia in approximately 90% of cases. Dedicated commercially available devices continue to evolve, each with its own advantages and limitations. Stent placement is subject to technical pitfalls, and adverse events occur following esophageal procedures in a minority of cases. Although chest pain is common and self-limited, reflux esophagitis, stent migration, tracheal compression, and esophageal perforation and obstruction require specific interventions. In many cases, these complications can be recognized and treated by the interventional radiologist with minimally invasive techniques. PMID- 12533646 TI - Hepatocellular carcinoma treated with radio-frequency ablation: spectrum of imaging findings. AB - Contrast material-enhanced Doppler or gray-scale harmonic ultrasonography (US) may help determine the completeness or long-term therapeutic efficacy of radio frequency (RF) ablation of hepatocellular carcinoma (HCC). Successfully treated HCC is devoid of vascularity at color or power Doppler US. When the tumor is not completely treated, residual viable tumor can be detected. These contrast enhanced US techniques may also help identify residual tumor when performed during repeat RF ablation, when accurate localization of viable tumor is needed. To date, contrast-enhanced computed tomography (CT) has been the most widely used imaging modality in the evaluation of therapeutic response after RF ablation of HCC. At follow-up CT, successfully ablated lesions appear as low-attenuation areas with no foci of contrast enhancement either within or at the periphery of the treated lesion, whereas any foci of enhancement indicate residual or recurrent tumor. Reactive hyperemia in tissue surrounding the ablated lesion, iatrogenic arterioportal shunting, and small intralesional air pockets are frequently seen at immediate follow-up CT. Gadolinium-enhanced dynamic magnetic resonance imaging is also useful in assessing therapeutic response following RF ablation of HCC, particularly when CT findings are inconclusive. Familiarity with these imaging findings is helpful in this setting. PMID- 12533647 TI - Major complications after radio-frequency thermal ablation of hepatic tumors: spectrum of imaging findings. AB - Although radio-frequency (RF) ablation has been accepted as a promising and safe technique for treatment of unresectable hepatic tumors, investigation of its complications has been limited. According to the multicenter (1,139 patients in 11 institutions) survey data of the Korean Study Group of Radiofrequency Ablation, a spectrum of complications occurred after RF ablation of hepatic tumors. The prevalence of major complications was 2.43%. The most common complications were hepatic abscess (0.66%), peritoneal hemorrhage (0.46%), biloma (0.20%), ground pad burn (0.20%), pneumothorax (0.20%), and vasovagal reflex (0.13%). Other complications were biliary stricture, diaphragmatic injury, gastric ulcer, hemothorax, hepatic failure, hepatic infarction, renal infarction, sepsis, and transient ischemic attack. One procedure-related death (0.09%) occurred (due to peritoneal hemorrhage). Three important strategies for decreasing the rate of complications are prevention, early detection, and proper management. A physician who performs RF ablation of hepatic malignancies should be aware of the broad spectrum of major complications so that these strategies can be used. PMID- 12533649 TI - Transvaginal US and hysterosonography in postmenopausal women with breast cancer receiving tamoxifen: correlation with hysteroscopy and pathologic study. AB - Tamoxifen citrate therapy increases the prevalence of benign and malignant uterine lesions. At transvaginal ultrasonography (US), the finding of a thickened central endometrial complex, with or without cystic changes, is often nonspecific and may be caused by an endometrial polyp, submucosal leiomyoma (fibroid), endometrial hyperplasia, carcinoma, or cystic atrophy. In addition, because of an increased prevalence of adenomyosis or adenomyosis-like changes in women receiving tamoxifen, proper transvaginal US assessment of endometrial thickness and abnormalities is difficult in some women. Hysterosonography, as an adjunct to transvaginal US, allows identification of intracavitary lesions and focal and diffuse endometrial abnormalities and helps determine whether an abnormality is endometrial or subendometrial. Endometrial polyps may be seen at transvaginal US as nonspecific thickening of the endometrial complex, with or without cystic changes. At hysterosonography, they appear as an echogenic mass with smooth margins. Submucosal leiomyomas may protrude into the endometrial cavity, causing false endometrial thickening at transvaginal US. Hysterosonography shows a round structure arising from the myometrium with a thin, overlying endometrium. At transvaginal US, when the endometrium cannot be accurately measured or when there is a nonspecific thickened central endometrial complex, hysterosonography can provide additional information and can help in the triage for hysteroscopic versus nondirected endometrial biopsy. Correlation of transvaginal US and hysterosonographic findings with hysteroscopic and pathologic findings enhances understanding of these changes, as well as the limitations and potential pitfalls of both imaging techniques. PMID- 12533651 TI - Cortical lesions of the tibia: characteristic appearances at conventional radiography. AB - Lesions that involve the cortex of the tibia are fairly common in radiology practice. However, the number of diseases that involve the tibial cortex is great, and it can be difficult to arrive at a limited differential diagnosis from radiographic findings. Categorization of lesions of the tibia into those that cause cortical destruction and those that cause cortical proliferation can help narrow the broad differential diagnosis. Lesions that cause cortical destruction include nonossifying fibroma, fibrous dysplasia, osteofibrous dysplasia, aneurysmal bone cyst, giant cell tumor, eosinophilic granuloma, Ewing sarcoma, neurofibromatosis, adamantinoma, osteoblastoma, chondromyxoid fibroma, hemangioendothelioma, renal cell metastatic disease, hemangioma, and hemangiopericytoma. Lesions that cause cortical proliferation include osteochondroma, stress fracture, osteoid osteoma, periosteal osteogenic sarcoma, diaphyseal dysplasia, venous stasis, cellulitis, chronic osteomyelitis, osteopathia striatum, and melorheostosis. Conventional radiography along with clinical and pathologic data can aid in diagnosis of the wide variety of disease processes that involve the tibial cortex. PMID- 12533652 TI - Discrimination of metastatic from acute osteoporotic compression spinal fractures with MR imaging. AB - A study was performed to determine which magnetic resonance (MR) imaging findings are useful in discrimination between metastatic compression fractures and acute osteoporotic compression fractures of the spine. The MR imaging findings in 27 patients with metastatic compression fractures and 55 patients with acute osteoporotic compression fractures were compared by using the chi(2) test. MR imaging findings suggestive of metastatic compression fractures were as follows: a convex posterior border of the vertebral body, abnormal signal intensity of the pedicle or posterior element, an epidural mass, an encasing epidural mass, a focal paraspinal mass, and other spinal metastases. MR imaging findings suggestive of acute osteoporotic compression fractures were as follows: a low signal-intensity band on T1- and T2-weighted images, spared normal bone marrow signal intensity of the vertebral body, retropulsion of a posterior bone fragment, and multiple compression fractures. The signal intensity on fast spin echo T2-weighted images obtained without fat suppression played little role in distinguishing between metastatic compression fractures and acute osteoporotic compression fractures. PMID- 12533653 TI - Management of spinal pain with imaging-guided injection. PMID- 12533654 TI - Special report: medical devices and foreign bodies: an introduction. PMID- 12533655 TI - Glossary of medical devices and procedures: abbreviations, acronyms, and definitions. PMID- 12533657 TI - From the archives of the AFIP: extratesticular scrotal masses: radiologic pathologic correlation. AB - The extratesticular scrotal contents consist of the epididymis, spermatic cord, and fascia derived from the embryologic descent of the testis through the abdominal wall. As opposed to intratesticular masses, most extratesticular masses are benign. Cystic masses (including hydroceles, epididymal cysts, and varicoceles) are easily diagnosed with ultrasonography (US) and are benign. Epididymitis is a common extratesticular lesion as well as the most frequent cause of an acute scrotum. It may be either acute or chronic and can be potentially complicated by epididymo-orchitis or scrotal abscess. Findings include epididymal enlargement, skin thickening, hydroceles, and hyperemia. The epididymis can also be affected by sarcoidosis, a noninfectious granulomatous disorder. The most common extratesticular neoplasms are lipomas (most often arising from the spermatic cord) and adenomatoid tumors (most often found in the epididymis). Despite their relative rarity, malignant neoplasms do occur and include rhabdomyosarcoma, liposarcoma, leiomyosarcoma, malignant fibrous histiocytoma, mesothelioma, and lymphoma. These tumors are often large at the time of presentation. The US findings of solid masses are often nonspecific. Magnetic resonance imaging can be very helpful in the evaluation of some of these disorders, allowing for a more specific diagnosis in cases of lipoma, fibrous pseudotumor, and polyorchidism. PMID- 12533658 TI - Best cases from the AFIP: angiomyolipomas in tuberous sclerosis. PMID- 12533659 TI - AFIP archives: gliomatosis cerebri affecting the entire neuraxis. PMID- 12533660 TI - Computer-aided diagnosis in chest radiography: results of large-scale observer tests at the 1996-2001 RSNA scientific assemblies. AB - Since 1996, computer-aided diagnosis (CAD) schemes have been presented as interactive demonstrations on computer workstations at each scientific assembly of the Radiological Society of North America. The schemes involved (a) detection of pulmonary nodules, (b) temporal subtraction, (c) detection of interstitial lung disease, (d) differential diagnosis of interstitial lung disease, and (e) distinction between benign and malignant pulmonary nodules on chest radiographs. Large-scale observer tests were carried out to examine how radiologists can benefit from CAD systems. Observer performance was evaluated by analysis of receiver operating characteristic (ROC) curves. The statistical significance of the difference between the areas under the ROC curves without and with CAD was analyzed with the Student t test. In all of the tests, the diagnostic accuracy of the radiologists in total improved significantly when CAD was used. This result provides additional evidence that CAD has the potential to improve the performance of radiologists in their decision-making process in interpreting chest radiographs. PMID- 12533661 TI - Use of personal digital assistants for retrieval of medical images and data on high-resolution flat panel displays. AB - For its new acute care hospital, the University of California at Los Angeles is evaluating innovative technology involving high-resolution flat panel display devices configured as "network appliances" that can be wall mounted for use in the retrieval and display of medical images and data. Physicians and healthcare providers can log on with wireless handheld computers, which can serve as an identification device as well as a navigational tool for selecting patient records and data. These data are displayed and manipulated on the flat panel display without the need for a keyboard or mouse. A prototype was developed with commercially available image display software, which was modified to allow the remote control of software functions from a handheld device through an infrared communication port. The system also allows navigation through the patient data in a World Wide Web-based electronic patient record. This prototype illustrates the evolution of radiologic facilities toward "shareable" high-quality display devices that allow more convenient and cost-effective access to medical images and related data in complex clinical environments, resulting in a paradigm shift in data navigation and accessibility. PMID- 12533663 TI - Real-Time Four-dimensional Imaging of the Heart with Multi-Detector Row CT. AB - An interactive four-dimensional (4D) visualizing system for the heart was developed by the authors. The system realizes high-resolution three-dimensional (3D) imaging with temporal resolution in a beating heart by using eight or more data sets reconstructed from multi-detector row computed tomography (MDCT) with a retrospective electrocardiograph-gated reconstruction algorithm. The motion of heart walls, papillary muscles, septa, and valves can now be observed in 4D multiplanar reformations (MPRs), as with sonography, while coronary arteries, coronary sinuses, and cardiac veins can be analyzed during the optimal phase in 4D volume-rendering images, as with angiography. All parameters such as window width, window level, field of view, panning, tilt, thresholds, opacity, color, and segmentation function are completely interactive in 4D imaging. Two longitudinal views and one latitudinal view of a heart can be simultaneously visualized in the three relative 4D MPR views. These newly developed capabilities in viewing both 3D volume and temporal resolution data, functional data, and even multiphase data with registration add considerable diagnostic potential. The advent of this real-time 4D visualizing system has enhanced the capabilities of MDCT. PMID- 12533665 TI - Sex pheromone biosynthetic pathway for disparlure in the gypsy moth, Lymantria dispar. AB - The pheromone biosynthetic pathway for production of the sex pheromone disparlure, 2-methyl-7R,8S-epoxy-octadecane, was determined for the gypsy moth. Each step in the pathway was followed by using deuterium-labeled compounds that could be identified by using GCMS. This approach provides unequivocal determination of specific reactions in the pathway. It was shown that the alkene precursor, 2-methyl-Z7-octadecene, is most likely made in oenocyte cells associated with abdominal epidermal cells. The pathway begins with valine contributing carbons for chain initiation, including the methyl-branched carbon, followed by chain elongation to 19 carbons. The double bond is introduced with an unusual Delta12 desaturase that utilizes a methyl-branched substrate. The resulting 18-methyl-Z12-nonadecenoate is decarboxylated to the hydrocarbon, 2 methyl-Z7-octadecene. The alkene is then transported to the pheromone gland through the hemolymph, most probably by lipophorin. At the pheromone gland, the alkene is unloaded and transformed into the epoxide disparlure for release into the environment. A chiral HPLC column was used to demonstrate that the (R,S) stereoisomer of the epoxide, (+)-disparlure is found in pheromone glands. PMID- 12533666 TI - A photochemical approach to the lipid accessibility of engineered cysteinyl residues. AB - Ordinary electrophilic reagents react too slowly in a nonpolar environment to be useful for the determination of the accessibility to lipid of continuous stretches of residues mutated to cysteine. By contrast, photoactivated 5 iodonaphthyl-1-azide (INA) reacted readily with 2-mercaptoethanol and dodecanethiol in nonpolar solvents and in liposomes. Continuous stretches of residues in the amphipathic N-terminal helix and first transmembrane helix of the bacterial potassium channel KcsA were replaced with cysteine, and the mutants were expressed in Escherichia coli and isolated in inner membranes. These membranes were dissolved in detergent and reconstituted into asolectin liposomes incorporating INA. The extent of light-induced reaction of INA with each cysteine was assayed by subsequent reaction with the gel-shifting, SH-specific methoxy polyethylene glycol-2-pyridine disulfide. The pattern of apparent second-order rate constants for the photoreactions of eight substituted cysteines in the N terminal helix conformed to other measures of lipid exposure. The pattern of the rate constants for the photoreactions of 15 cysteines in the first transmembrane helix had peaks every third residue, which partly conformed to other measures of lipid exposure. PMID- 12533667 TI - Potential involvement of the cyclooxygenase-2 pathway in the regulation of tumor associated angiogenesis and growth in pancreatic cancer. AB - Angiogenesis plays a crucial role in tumor development and growth. The present investigation was undertaken to test the potential involvement of the cyclooxygenase-2 (COX-2) pathway in the regulation of angiogenesis and growth in pancreatic cancer. We compared the angiogenic characteristics of a COX-2-positive human pancreatic tumor cell line, BxPC-3, with those of a COX-2-negative pancreatic tumor cell line, AsPC-1. Cultured BxPC-3 cells promoted a marked increase of endothelial cell migration in comparison with migration that occurred in the absence of cancer cells. Furthermore, BxPC-3 cell culture supernatants induced endothelial cell capillary morphogenesis in vitro and neovascularization in vivo. In contrast, cultured AsPC-1 cells elicited a modest effect on endothelial cell migration and neovascularization in vivo. Pretreatment of BxPC-3 cells with the selective COX-2 inhibitor NS-398 (50 micro M) dramatically decreased angiogenic responses of endothelial cells. NS-398 (25-100 micro M) caused inhibition of BxPC-3 cell proliferation but had no effect on AsPC-1 cell growth. SC-560, a selective COX-1 inhibitor, had no effect on growth of either cell lines. These results suggest an involvement of COX-2 in the control of tumor dependent angiogenesis and growth in certain pancreatic cancers and provide the rational for inhibition of the COX pathway as an effective therapeutic approach for pancreatic tumors. PMID- 12533668 TI - Negative association of melanoma differentiation-associated gene (mda-7) and inducible nitric oxide synthase (iNOS) in human melanoma: MDA-7 regulates iNOS expression in melanoma cells. AB - The melanoma differentiation association gene-7 (mda-7) is a novel tumor suppressor gene of which the protein expression decreases to nearly undetectable levels in metastatic melanoma. In contrast, expression of inducible nitric oxide synthase (iNOS) is increased in advanced stages of melanoma, and iNOS expression has been proposed as a potential prognostic marker in this disease. Thus, expression of these molecules in the same tumor appears to exhibit reciprocal characteristics. We hypothesize that the relative ratios of these melanoma progression molecules may define either tumor progression or tumor suppression in human melanoma. The first goal of this study was to determine whether MDA-7 expression in melanoma negatively correlates with iNOS expression. The second goal was to determine whether iNOS expression could be regulated by MDA-7 expression in melanoma cells. Expression of MDA-7 and iNOS proteins were evaluated by immunohistochemistry in a total of 81 tumor samples: 38 primary melanomas and 43 metastatic melanomas. Evaluation of these melanoma patient samples showed that expression of MDA-7 and iNOS exhibits a significant negative correlation (P = 0.03). In vitro studies revealed that iNOS expression in melanoma cell lines is lost in a dose-dependent fashion after treatment with an adenoviral vector encoding the mda-7 gene (Ad-mda7) or with rhMDA-7 protein, demonstrating that MDA-7 down-regulates iNOS expression. Furthermore, we demonstrate that the STAT-3-modulated expression of IFN regulatory factors 1 and 2 is regulated by MDA-7, which may alter iNOS gene expression. These studies demonstrate that expression of the MDA-7 tumor suppressor can negatively regulate iNOS expression in malignant melanoma cell lines. PMID- 12533669 TI - A novel form of the plasminogen activator inhibitor created by cysteine mutations extends its half-life: relevance to cancer and angiogenesis. AB - Proteolytic activity driven by urokinase (uPA) is commonly recognized as an important factor in metastasis and angiogenesis. The eradication of unwanted uPA activity expressed by cancer cells results in the inhibition of metastasis and angiogenesis. Development of novel and highly selective uPA inhibitors could, therefore, produce new treatments of cancer. The ultimate goal of this work is the identification of novel and selective inhibitors of uPA suitable for this purpose. We have chosen plasminogen activator inhibitor(s) type 1 (PAI-1), which selectively inhibits the urokinase plasminogen activator (uPA). However, PAI-1 is not a stable molecule and converts itself into the latent form with a half-life in the range of t1/2 = 1-2 h. This conversion is associated with a partial insertion of the reactive loop (P4-P10') into the PAI-1 molecule. In such a conformation, P1-P1' and other sites are not accessible for reaction with uPA. To conquer this hurdle, we have produced several PAI-1 mutants by replacing chosen amino acids with cysteine in the hope of creating disulfide bridges, which could make this insertion more difficult. On the basis of the known structure of active PAI-1, we have identified amino acids that can be substituted with a cysteine residue to produce disulfide bridges linking the top and bottom parts of strands A3 and A5 as well as sites within the helix-D region. We created a total of seven cysteine mutants via point mutation (two to six point mutations), generating possible sites for disulfide bridge formation at the top and bottom parts of A3 and A5, within the helix-D region, or by a combination thereof. Desired mutations were introduced by PCR using appropriate primers. The mutant forms of PAI-1 containing the chitin-binding intein tag were then purified using affinity chromatography wherein the intein tag is cleaved, leaving mutant PAI-1 protein. Cys mutations resulted in proteins with extended half-life of PAI-1 from 2 to >700 h depending on the mutant. Novel PAI-1 were fully functional against uPA and showed activity in the in vitro model of angiogenesis, e.g., in the inhibition of sprout formation. Such prolonged serpin activity, which is therapeutically desired in cancer treatment and Cys-mutated PAI-1, could launch a new class of novel anticancer agents. PMID- 12533670 TI - Camptothecin poly[n-(2-hydroxypropyl) methacrylamide] copolymers in antitopoisomerase-I tumor therapy: intratumor release and antitumor efficacy. AB - Soluble copolymers of camptothecin (CPT), based on poly[N-(2-hydroxypropyl) methacrylamide] (pHPMA), were obtained by conjugation through the degradable spacers -Gly-Phe-Leu-Gly- or -Gly-6-aminohexanoyl-Gly-. We investigated to what extent passive accumulation and retention of hydroxypropyl methacrylamide copolymer of CPT (pHPMA-CPT) in tumors and modulation of the drug release influence efficacy. Release of CPT in vivo was detected by time-resolved phase shift fluorescence imaging on tumor specimens, based on the evidence that free and bound drug had different fluorescence lifetimes in solution. HT-29 murine specimens, obtained at several times after treatment with (3)H-labeled free CPT, pHPMA-Gly-Phe-Leu-Gly-CPT, or pHPMA-Gly-6-aminohexanoyl-Gly-CPT, were either imaged for time-resolved phase-shift fluorescence or subjected to autoradiography. Phase shifts of CPT conjugates were equal or longer than those of free CPT, indicating the presence of both free and polymer-bound drug in the tumor, in agreement with autoradiograms. pHPMA-Gly-Phe-Leu-Gly-CPT underwent relevant intratumor hydrolysis during the first 24 h, whereas the hydrolysis of pHPMA-Gly-6-aminohexanoyl-Gly-CPT was slow. The latter showed antitumor activity at doses from 10 to 22.5 mg/kg/day against s.c. HT-29, A2780, M14, and A549 s.c. xenografts. Moreover, inhibition of tumor growth lasted for up to 73-88 days, and cures were observed on mice with orthotopic implanted HT-29; pHPMA-Gly-Phe-Leu Gly-CPT was 2-fold more potent than pHPMA-Gly-6-aminohexanoyl-Gly-CPT but less tolerated. Our data suggest that the efficacy of pHPMA-CPT copolymers is related to their intratumor accumulation, and in vivo properties of releasing CPT by esterolytic and proteolytic degradation. PMID- 12533671 TI - Induction of DNA damage responses by adozelesin is S phase-specific and dependent on active replication forks. AB - Adozelesin is an alkylating minor groove DNA binder that is capable of rapidly inhibiting DNA replication in treated cells through a trans-acting mechanism and preferentially arrests cells in S phase. It has been shown previously that in cells treated with adozelesin, replication protein A (RPA) activity is deficient, and the middle subunit of RPA is hyperphosphorylated. The adozelesin-induced RPA hyperphosphorylation can be blocked by the replicative DNA polymerase inhibitor, aphidicolin, suggesting that adozelesin-triggered cellular DNA damage responses require active DNA replication forks. These data imply that cellular DNA damage responses to adozelesin treatment are preferentially induced in S phase. Here, we show that RPA hyperphosphorylation, RPA intranuclear focalization, and gamma-H2AX intranuclear focalization induced by adozelesin treatment are all dependent on DNA replication fork progression, and focalization is only induced in S phase cells. These findings are similar to those seen with the S phase-specific DNA damaging agent, camptothecin. Conversely, all three DNA damage responses are independent of either S phase or replication fork progression when induced by treatment with the DNA strand scission agent, C-1027. Furthermore, we demonstrate that adozelesin-induced RPA and gamma-H2AX intranuclear foci appear to colocalize within the nuclei of S phase cells. PMID- 12533672 TI - Clinically potential subclasses of retinoid synergists revealed by gene expression profiling. AB - Retinoids have chemopreventive and therapeutic potency in oncology and dermatology, although their application is restricted by many undesirable side effects. For the development of more effective and less toxic retinoids, gene expression analyses using DNA microarrays have the potential to supplement conventional screening methods, which are based on the changes in cell morphology and/or function. In this study, we applied the class prediction algorithm, which was used in the molecular phenotyping of tumors, for the classification of synthetic retinoids (Am80 and Tp80) and retinoid synergists (HX630, TZ335, and PA024) as all-trans retinoic acid-like, 9-cis retinoic acid-like, and control like classes. By analyzing the effects of all-trans retinoic acid and 9-cis retinoic acid on the gene expressions in a human promyelocytic leukemia cell line, HL60, we successfully selected 50 marker genes whose expression pattern could distinguish these classes. Moreover, the classification revealed the existence of two subclasses among the retinoid synergists used with Am80. Close inspection of the DNA microarray analyses indicated that these two subclasses had different effects on the apoptosis of HL60 cells, and this was confirmed by in vivo experiments. These results indicate that the retinoidal activity of Am80, which has already been used in clinical trials, could be modulated differently by the two classes of retinoid synergists. Thus, these two subclasses of retinoid synergists have the potency to widen the usage of Am80. Our analyses demonstrated that the gene expression profiling could provide important information for developing useful retinoid synergists by compensating conventional screening methods. PMID- 12533673 TI - Effects of ER-37328 on primary tumor, liver metastasis, and life span in a murine colon 38 orthotopic transplantation model. AB - 12,13-Dihydro-5-[2-(dimethylamino)ethyl]-4H-benzo[c] pyrimido[5,6,1-jk] carbazole 4,6,10(5H,11H)-trione hydrochloride (ER-37328) is a novel topoisomerase II poison with potent tumoricidal activity against solid tumor cells both in vitro and in vivo. Here, we describe studies on the effects of ER-37328 on the primary tumor, liver metastasis, and survival in a murine Colon 38 orthotopic transplantation model. When ER-37328 (10 mg/kg) was administered i.v. at 11 days or 20 days after transplantation, strong regression of the primary tumor was observed on both administration schedules. On the later schedule, ER-37328 completely blocked liver metastasis, whereas the mean number of metastases in the control group was 23.9. To examine the antitumor activity against Colon 38 at the liver in more detail, ER-37328 was administered to mice that had received an inoculation of Colon 38 tumor into the liver. ER-37328 showed strong tumor-regression activity against Colon 38 growing in the liver. In addition, administration of ER-37328 on a schedule of every 7 days four times caused a significant increase of 79% in life span in the orthotopic transplantation model, calculated by using mean survival times. Pharmacokinetic study revealed that ER-37328 was highly distributed to the tumor and organs. The ratios of the area under the concentration-time curves of ER-37328 in the tumor, lung, liver, and kidney versus plasma were 81, 77, 47, and 40, respectively. This high distribution to the tumor and liver may explain the potent antitumor activity of ER-37328 against Colon 38 tumor in the liver. In conclusion, the topoisomerase II poison ER-37328 is a promising candidate for clinical application against colon cancer. PMID- 12533674 TI - Triptolide inhibits the growth and metastasis of solid tumors. AB - Triptolide (TPL), a diterpenoid triepoxide purified from the Chinese herb Tripterygium wilfordii Hook F, was tested for its antitumor properties in several model systems. In vitro, TPL inhibited the proliferation and colony formation of tumor cells at extremely low concentrations (2-10 ng/ml) and was more potent than Taxol. Likewise, in vivo, treatment of mice with TPL for 2-3 weeks inhibited the growth of xenografts formed by four different tumor cell lines (B16 melanoma, MDA 435 breast cancer, TSU bladder cancer, and MGC80-3 gastric carcinoma), indicating that TPL has a broad spectrum of activity against tumors that contain both wild type and mutant forms of p53. In addition, TPL inhibited experimental metastasis of B16F10 cells to the lungs and spleens of mice. The antitumor effect of TPL was comparable or superior with that of conventional antitumor drugs, such as Adriamycin, mitomycin, and cisplatin. Importantly, tumor cells that were resistant to Taxol attributable to the overexpression of the multidrug resistant gene 1 were still sensitive to the effects of TPL. Studies on cultured tumor cells revealed that TPL induced apoptosis and reduced the expression of several molecules that regulate the cell cycle. Taken together, these results suggest that TPL has several attractive features as a new antitumor agent. PMID- 12533675 TI - Flavopiridol-induced apoptosis is mediated through up-regulation of E2F1 and repression of Mcl-1. AB - Flavopiridol treatment can lead to apoptosis via a mechanism that has been associated with down-regulation of Mcl-1. Likewise, recent studies from our laboratory demonstrated that E2F1 leads to transcriptional repression of Mcl-1 and subsequently apoptosis. Given the ability of cyclin/cyclin-dependent kinase 2 antagonists to kill transformed cells, we surmised that flavopiridol may stabilize E2F1 and enhance apoptosis via repression of Mcl-1. Here we demonstrate that flavopiridol is associated with a dose-dependent increase in E2F1 protein levels, a corresponding reduction in Mcl-1, and apoptosis in H1299 lung carcinoma cells. Treatment of H1299 cells with 200 nM flavopiridol resulted in the rapid elevation of E2F1 and reduction in Mcl-1 levels within 12 h of treatment. The elevation of E2F1 and reduction in Mcl-1 clearly preceded the induction of apoptosis. Both H1299 and NIH3T3 fibroblast cell lines that constitutively express Mcl-1 under the control of the cytomegalovirus promoter have no reductions in Mcl-1 levels with flavopiridol treatment and are resistant to apoptosis induced by flavopiridol. H1299 cells that have E2F1 deleted through RNAi vector targeting are less sensitive to flavopiridol-induced cell death, and likewise, mouse embryo fibroblast cell lines deficient in E2F1 are less susceptible to apoptosis induced by flavopiridol compared with wild-type control fibroblasts. These data suggest that apoptosis induced by flavopiridol is dependent on the enhancement of E2F1 levels and the repression of Mcl-1. PMID- 12533676 TI - Protein kinase C-dependent activation of the tumor necrosis factor receptor mediated extrinsic cell death pathway underlies enhanced apoptosis in human myeloid leukemia cells exposed to bryostatin 1 and flavopiridol. AB - Interactions between the protein kinase C activator bryostatin 1 and the cyclin dependent kinase (CDK) inhibitor flavopiridol (FP) have been examined in human myeloid leukemia cells (U937 and HL-60). Previous studies have demonstrated synergistic induction of apoptosis in leukemic cells exposed to the potent differentiation-inducer phorbol 12-myristate 13-acetate (PMA) in conjunction with FP [L. Cartee et al., Cancer Res., 61: 2583-2591, 2001]. Although bryostatin 1 (10 nM) is a very weak inducer of differentiation compared with PMA in these cells, coadministration of a minimally toxic concentration of FP (100 nM) did not promote bryostatin 1-related maturation but instead caused a marked increase in mitochondrial damage (e.g., cytochrome c release; loss of Deltapsi(m)), caspase activation, poly(ADP-ribose) polymerase cleavage, and apoptosis. Bryostatin 1/FP induced apoptosis was significantly diminished in cells ectopically expressing dominant-negative Fas-associated death domain or by coadministration of tumor necrosis factor (TNF)-alpha soluble receptors, implicating the extrinsic pathway in bryostatin 1/FP actions. Enhanced apoptosis in bryostatin 1/FP-treated cells was accompanied by down-regulation of Mcl-1 and a sustained increase in TNF-alpha release. The selective protein kinase C inhibitor GFX blocked TNF-alpha and cytochrome c release in bryostatin 1/FP-treated cells and attenuated apoptosis. Finally, coadministration of bryostatin 1 (or PMA) with FP induced a marked increase in apoptosis in U937 cells ectopically expressing an NH(2)-terminal phosphorylation loop-deleted Bcl-2 protein, which are otherwise highly resistant to FP-mediated lethality. Taken together, these findings suggest that synergistic induction of apoptosis by bryostatin 1 and FP does not stem from disruption of the leukemic cell maturation process but instead results from enhanced release of TNF-alpha and activation of the extrinsic apoptotic cascade, culminating in cell death. PMID- 12533677 TI - Curcumin (diferuloyl-methane) enhances tumor necrosis factor-related apoptosis inducing ligand-induced apoptosis in LNCaP prostate cancer cells. AB - The role of natural food products in prevention of prostate cancer has been confirmed in recent epidemiological studies; however, the mechanism of chemoprevention by the dietary constituents largely remains unknown. Curcumin, the yellow pigment and active component of turmeric (Curcuma longa), exhibits chemopreventive and growth inhibitory activity against several tumor cell lines. The androgen-sensitive human prostate cancer cell line LNCaP is only slightly susceptible to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), a member of the tumor necrosis factor family of cell death-inducing ligands. In this study, we investigated whether curcumin and TRAIL cooperatively interact to promote death of LNCaP cells. At low concentrations (10 micro M curcumin and 20 ng/ml TRAIL), neither of the two agents alone produced significant cytotoxicity (curcumin, <10%; TRAIL, approximately 15%) in LNCaP cells, as measured by the 3 (4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfonyl)-2H tetrazolium dye reduction assay. On the other hand, cell death was markedly enhanced (2-3-fold) if tumor cells were treated with curcumin and TRAIL together. The combined curcumin and TRAIL treatment increased the number of hypodiploid cells and induced DNA fragmentation in LNCaP cells. The combined treatment induced cleavage of procaspase-3, procaspase-8, and procaspase-9, truncation of Bid, and release of cytochrome c from the mitochondria, indicating that both the extrinsic (receptor-mediated) and intrinsic (chemical-induced) pathways of apoptosis are triggered in prostate cancer cells treated with a combination of curcumin and TRAIL. These results define a potential use of curcumin to sensitize prostate cancer cells for TRAIL-mediated immunotherapy. PMID- 12533678 TI - Reversal of breast cancer resistance protein-mediated drug resistance by estrogen antagonists and agonists. AB - Breast cancer resistance protein (BCRP), an ATP-binding cassette transporter, confers resistance to a series of anticancer agents such as SN-38, mitoxantrone, and topotecan. In a previous study, we found that estrogens reverse drug resistance of BCRP-expressing cells. In this study, estrogen antagonists, estrogen agonists, and their derivatives were evaluated for BCRP-reversing activity. First, compounds were tested for effects on the cellular accumulation of topotecan in BCRP-transduced K562 cells (K562/BCRP). Next, these compounds were examined for their ability to reverse SN-38 and mitoxantrone resistance in K562/BCRP cells. Among commercially available estrogen antagonists and agonists tested, diethylstilbestrol showed the strongest BCRP-reversing activity. Diethylstilbestrol increased the cellular accumulation of topotecan and reversed drug resistance in K562/BCRP cells but showed marginal or no effect in parental K562 cells. The reversal activities of estrone and diethylstilbestrol were more prominent for mitoxantrone than for SN-38. Tamoxifen and toremifene were also found to enhance topotecan uptake in K562/BCRP cells. Next, various tamoxifen derivatives were screened for anti-BCRP activity. In the first cycle of screening with 14 compounds, TAG-11 showed the strongest effect. In the second cycle of screening of 25 TAG-11-related compounds, TAG-139 showed the strongest effect. Reversal of SN-38 and mitoxantrone resistance in K562/BCRP cells by TAG-139 was 5 fold stronger than that by estrone. Dose-dependent characteristics of drug resistance reversal with estrone and TAG-139 were very similar, suggesting that estrone and tamoxifen derivatives interact with the same drug-binding site of BCRP. Derivatives of antiestrogens that exhibit no other biological effects promise to be useful in overcoming BCRP-mediated drug resistance. PMID- 12533679 TI - Identification of the polycomb group protein SU(Z)12 as a potential molecular target for human cancer therapy. AB - We have previously identified SU(Z)12 as an E2F target gene. Because many E2F target genes encode proteins that are critical for the control of cell proliferation, we have further characterized the regulation and expression of SU(Z)12. To understand the molecular mechanisms responsible for expression of SU(Z)12 mRNA, we have analyzed the promoter region. We found that the SU(Z)12 gene is controlled by dual promoters, one of which functions bidirectionally. In addition to the E2F binding site, we have identified two binding sites for T cell factor (TCF)/beta-catenin complexes. Using gel mobility shift assays, we demonstrated that both TCF sites can be bound by TCF4. TCF/beta-catenin complexes have been shown to be a critical regulator of gene expression in tumors of the colon, breast, and liver. Accordingly, we have used chromatin immunoprecipitation assays to confirm that TCF4/beta-catenin complexes are bound to the SU(Z)12 promoter in colon cancer cells but not in HeLa cells. We next adapted the chromatin immunoprecipitation assay for use with primary colon tumor samples, and, using matched pairs of normal and tumor tissue obtained from several different colon cancer patients, we demonstrate that levels of beta-catenin bound to the SU(Z)12 promoter are increased in colon tumors. Finally, we show that the SU(Z)12 mRNA is up-regulated in a number of different human tumors, including tumors of the colon, breast, and liver. Recent studies have found that SU(Z)12 is a component of the Drosophila ESC-E(Z) and the human EED-EZH2 Polycomb chromatin remodeling complexes. Therefore, we suggest that SU(Z)12, which may modulate the tumor phenotype by changing gene expression profiles, may be a logical target for the design of a new antitumor agent PMID- 12533681 TI - Apoptosis and apoptosis-related molecules in the submandibular gland of the nonobese diabetic mouse model for Sjogren's syndrome: limited role for apoptosis in the development of sialoadenitis. AB - Sjogren's syndrome is an autoimmune disease in which lymphocytic infiltrates develop in the exocrine glands. Pathogenetic aspects of the disease can be studied in the nonobese diabetic (NOD) mouse strain, a spontaneous model for Sjogren's syndrome. Apoptosis may play a role in the initation phase and in the effector phase of autoimmune diseases. Here, we have examined the role of apoptosis in the development of sialoadenitis in the NOD mouse. Apoptotic cells and the expression of apoptosis-related molecules were studied in submandibular glands (SMG) of NOD and NOD-scid mice before and after the onset of sialoadenitis. Numbers of apoptotic cells were not increased as compared with control mice, at any age. By immunohistochemistry, we demonstrated increased expression of Fas, Fas ligand (FasL), and bcl-2 on SMG epithelial cells of NOD and NOD-scid mice, as early as 3 days of age. mRNA expression of Fas and FasL was also examined in SMG by RQ-PCR. Low-level expression of Fas and FasL mRNA was observed in all mouse strains, from 1 day of age onward. We conclude that increased protein expression of Fas and FasL on SMG epithelial cells of NOD and NOD-scid mice probably indicates a genetically programmed abnormality in these cells that may form a trigger for the development of sialoadenitis in NOD mice. Because increased numbers of apoptotic cells were not observed, a role for actual apoptosis in the initiation or effector phase of sialoadenitis in the NOD mouse is unlikely. PMID- 12533682 TI - Increased levels of low-density lipoprotein oxidation in patients with familial hypercholesterolemia and in end-stage renal disease patients on hemodialysis. AB - Patients with familial hypercholesterolemia (FH) and patients with end-stage renal disease (ESRD) undergoing dialysis suffer from accelerated atherosclerosis. Oxidation of low-density lipoprotein (LDL) cholesterol is crucial in atherogenesis. In the present study, we determined the LDL oxidation level and oxidizability of isolated LDL of 11 male patients with FH, 15 male ESRD patients on hemodialysis, and 15 age-matched male normolipidemic healthy controls. FH patients were without lipid-lowering medication for at least 4 weeks and were reassessed after 2 years of cholesterol-lowering therapy (statins). LDL oxidation level was measured by ELISA using monoclonal antibody 4E6 to oxidized LDL (oxLDL) as the capture antibody and anti-human apoB antibody for detection; results were expressed as percentage oxLDL. In FH patients and in ESRD patients on hemodialysis, both groups having a higher percentage of cardiovascular disease, mean plasma LDL oxidation levels were significantly elevated compared with controls (4.9 +/- 1.3; 3.7 +/- 2.0; 1.7 +/- 0.6%, respectively). Within each group of subjects, LDL oxidation level was not associated with history of cardiovascular disease. Furthermore, in neither group was a significant correlation found between plasma concentration of LDL cholesterol and LDL oxidation level. After cholesterol-lowering therapy, LDL oxidation level in FH patients had not changed significantly and remained elevated compared with controls, despite a reduction of LDL cholesterol by 55% on average. Also, absolute plasma oxLDL concentrations, obtained by multiplying LDL oxidation level with plasma LDL cholesterol concentration, were significantly higher in FH patients before and after cholesterol-lowering therapy and in ESRD patients on hemodialysis than in controls (489 +/- 145; 189 +/- 122; 100 +/- 65; and 59 +/- 27 micro moles/L, respectively). No correlation was found between plasma oxLDL concentration and parameters of LDL oxidizability, LDL fatty acids, and LDL alpha tocopherol content. We conclude that cholesterol-lowering therapy does not normalize elevated LDL oxidation levels in FH patients and elevated LDL oxidation level in FH and in ESRD might mirror atherosclerosis. PMID- 12533683 TI - The CXC chemokine GCP-2/CXCL6 is predominantly induced in mesenchymal cells by interleukin-1beta and is down-regulated by interferon-gamma: comparison with interleukin-8/CXCL8. AB - Human granulocyte chemotactic protein-2 (GCP-2)/CXCL6 is a CXC chemokine that functionally uses both of the IL-8/CXCL8 receptors to chemoattract neutrophils but that is structurally most related to epithelial cell-derived neutrophil attractant-78 (ENA-78)/CXCL5. This study provides the first evidence that GCP-2 protein is, compared with IL-8, weakly produced by some sarcoma, but less by carcinoma cells, and is tightly regulated in normal mesenchymal cells. IL-1beta was the predominant GCP-2 inducer in fibroblasts, chondrocytes, and endothelial cells, whereas IL-8 was equally well up-regulated in these cells by TNF-alpha, measles virus, or double-stranded RNA (dsRNA). In contrast, lipopolysaccharide (LPS) was a relatively better stimulus for GCP-2 versus IL-8 in fibroblasts. IFN gamma down-regulated the GCP-2 production in fibroblasts induced by IL-1beta, TNF alpha, LPS, or dsRNA. The kinetics of GCP-2 induction by IL-1beta, LPS, or dsRNA in fibroblasts differed from those of IL-8. Freshly isolated peripheral blood mononuclear leukocytes, which are a good source of IL-8 and ENA-78, failed to produce GCP-2. However, lung macrophages and blood monocyte-derived macrophages produced GCP-2 in response to LPS. Quantitatively, secretion of GCP-2 always remained inferior to that of IL-8, despite the fact that the ELISA recognized all posttranslationally modified GCP-2 isoforms. The expression of GCP-2 was confirmed in vivo by immunohistochemistry. The patterns of producer cell types, inducers and kinetics and the quantities of GCP-2 produced, suggest a unique role for GCP-2 in physiologic and pathologic processes. PMID- 12533684 TI - Mutations in the STK11 gene characterize minimal deviation adenocarcinoma of the uterine cervix. AB - Minimal deviation adenocarcinoma (MDA) is a well-differentiated variant of mucinous adenocarcinoma of the uterine cervix and is found relatively infrequently in the general population. However, MDA is strongly associated with Peutz-Jeghers syndrome (PJS), a rare hereditary autosomal disorder characterized by benign hamartomatous polyposis in the gastrointestinal tract and mucocutaneous pigmentation. A serine threonine kinase gene, STK11, has been identified as the tumor suppressor gene responsible for the PJS. In this study we investigated the possible direct role of STK11 in the development of MDA of the uterine cervix. Eleven rare cases of mucinous MDA, not known to be associated with PJS, were screened for the presence of mutations in the STK11 gene by single-strand conformation polymorphism analysis of PCR-amplified DNA fragments. Subsequently our findings were confirmed with cloning and sequencing. As a control, 24 cases of endocervical adenocarcinomas of other histologic subtypes, with no family history of PJS (19 mucinous adenocarcinomas, 4 endometrioid adenocarcinomas, and 1 clear cell adenocarcinoma), 15 cases of squamous cell carcinomas of the uterine cervix, 5 cases of endocervical glands with pyloric gland metaplasia, and 2 deeply situated nabothian cysts were investigated. Somatic mutations of the STK11 gene were confirmed in 6 (55%) of the 11 mucinous MDAs and 1 (5%) of the 19 mucinous adenocarcinomas, but not in the 5 nonmucinous adenocarcinomas, the 15 squamous cell carcinomas, nor the 5 endocervical glands with gastric metaplasia. MDAs with the STK11 mutation had a significantly poorer prognosis than MDAs without the STK11 mutation (p = 0.039). A germline mutation of STK11 was detected in one PJS patient with mucinous adenocarcinoma of the uterine cervix. These results suggest that mutations in the STK11 gene may play an important role in the etiology of MDA of the uterine cervix and may distinguish this rare tumor from other common types of adenocarcinoma of the uterine cervix. PMID- 12533685 TI - PYY-Tag transgenic mice displaying abnormal (H+-K+)ATPase activity and gastric mucosal barrier impairment. AB - The mechanism by which the gastrointestinal hormones peptide YY and glucagon inhibit gastric acid secretion is largely unknown. PYY-Tag transgenic mice develop endocrine tumors in the colon that are composed mainly of peptide YY/enteroglucagon-producing L type cells. Therefore we studied the functional activity of such tumors and the gastric functions of PYY-Tag mice. Fasting and fed PYY-Tag transgenic mice and CD1 controls were assayed for circulating levels of peptide YY, glucagon, insulin, and gastrin. The gastric pH was determined and gastric samples were examined for (a) histologic appearance; (b) K(+)-stimulated p-nitrophenylphosphatase activity and [(14)C]aminopyrine accumulation of apical and tubulovesicle membranes; (c) adherent mucus determination by Alcian blue recovery; and (d) DNA/RNA/protein epithelial content and in vivo incorporation of [(3)H]thymidine into DNA. Transgenic mice showed high serum levels of peptide YY and glucagon, increased gastric pH, and a high incidence of gastric ulcers after fasting. p-Nitrophenylphosphatase activity, [(14)C] aminopyrine accumulation, and proton pump redistribution from cytoplasmic tubulovesicles to apical membranes were significantly lower in the gastric mucosa of transgenic mice compared with the controls. In addition, the adherent mucus was thinner, and [(3)H]thymidine incorporation into the DNA was decreased. The abnormal and unregulated levels of circulating peptide YY and glucagon led to gastric acid inhibition and an impairment of gastric barrier function as a result of a striking reduction in epithelial proliferation. PMID- 12533686 TI - Presence of restricted killer immunoglobulin-like receptor repertoire and monoclonal T-cell receptor gamma rearrangement as evidence of mixed NK/T-cell differentiation in a subset of sinonasal lymphomas. AB - Most sinonasal lymphomas have a restricted killer immunoglobulin-like receptor (KIR) repertoire without a monoclonal T-cell receptor-gamma (TCR-gamma) rearrangement, implying an NK lineage. However, the lineage assignment of sinonasal lymphoma with a monoclonal TCR-gamma rearrangement is unclear because of its mixed NK/T phenotype. The possibility of a mixed NK/T lineage arises with the discovery of T cells with NK features, such as KIR(+) T cells or Valpha24(+) NKT cells. The former might transform into a T-cell lymphoma with both a monoclonal TCR-gamma rearrangement and a restricted KIR repertoire; the latter might give rise to a T-cell lymphoma with a monoclonal Valpha24 rearrangement and possibly a restricted KIR repertoire. To identify such mixed-lineage lymphomas, we undertook a survey of 15 consecutive sinonasal lymphomas and found six with both a restricted KIR repertoire and a monoclonal TCR-gamma rearrangement, consistent with KIR(+) T-cell lymphomas. Among these six cases, four female CD56( )/CD44(-)/CD8(-)/CD45RO(+)/CD45RA(-) cases constituted a distinct group with a better prognosis than the rest of the male cases of sinonasal lymphomas. None of the six cases had a monoclonal Valpha24 repertoire, thus excluding a derivation from NKT cells. The predominance of KIR(+) T cells that normally function in chronic viral infections over Valpha24(+) NKT cells that typically recognize glycolipid antigens is consistent with the known association of Epstein-Barr virus infection with sinonasal lymphoma. The demonstration of mixed lineage in a mature lymphoid neoplasm is unusual and echoes the World Health Organization classification that placed NK-cell and T-cell lymphomas in a mixed group. PMID- 12533687 TI - Hypoxic preconditioning augments efficacy of human endothelial progenitor cells for therapeutic neovascularization. AB - A subset of human peripheral blood mononuclear cells (PB-MNCs) differentiate into endothelial progenitor cells (EPCs) that participate in postnatal neovascularization. Although tissue ischemia can mobilize EPCs from bone marrow, the effects of hypoxia on differentiation and angiogenic function of EPCs are little known. We examined whether hypoxic conditioning would modulate differentiation and function of human PB-MNC-derived EPCs. A subset of PB-MNCs gave rise to EPC-like attaching (AT) cells under either normoxic or hypoxic conditions. However, hypoxia much enhanced the differentiation of AT cells from PB-MNCs compared with normoxia. AT cells released vascular endothelial growth factor (VEGF) protein and expressed CD31 and kinase insert domain receptor/VEGFR 2, endothelial lineage markers, on their surface, which were also enhanced by hypoxia. Both a neutralizing anti-VEGF mAb and a KDR-specific receptor tyrosine kinase inhibitor, SU1498, suppressed PB-MNC differentiation into EPC-like AT cells in a dose-dependent manner. Migration of AT cells in response to VEGF as examined by a modified Boyden chamber apparatus was also enhanced by hypoxia. Finally, in vivo neovascularization efficacy was significantly enhanced by in vitro hypoxic conditioning of AT cells when cells were transplanted into the ischemic hindlimb of immunodeficient nude rats. In conclusion, hypoxia directly stimulated differentiation of EPC-like AT cells from human PB-MNC culture. Moreover, hypoxic preconditioning of AT cells before in vivo transplantation is a useful means to enhance therapeutic vasculogenesis. PMID- 12533688 TI - Coxsackievirus B3-associated myocardial pathology and viral load reduced by recombinant soluble human decay-accelerating factor in mice. AB - Coxsackievirus B3 (CVB3) infection can result in myocarditis, which in turn may lead to a protracted immune response and subsequent dilated cardiomyopathy. Human decay-accelerating factor (DAF), a binding receptor for CVB3, was synthesized as a soluble IgG1-Fc fusion protein (DAF-Fc). In vitro, DAF-Fc was able to inhibit complement activity and block infection by CVB3, although blockade of infection varied widely among strains of CVB3. To determine the effects of DAF-Fc in vivo, 40 adolescent A/J mice were infected with a myopathic strain of CVB3 and given DAF-Fc treatment 3 days before infection, during infection, or 3 days after infection; the mice were compared with virus alone and sham-infected animals. Sections of heart, spleen, kidney, pancreas, and liver were stained with hematoxylin and eosin and submitted to in situ hybridization for both positive strand and negative-strand viral RNA to determine the extent of myocarditis and viral infection, respectively. Salient histopathologic features, including myocardial lesion area, cell death, calcification and inflammatory cell infiltration, pancreatitis, and hepatitis were scored without knowledge of the experimental groups. DAF-Fc treatment of mice either preceding or concurrent with CVB3 infection resulted in a significant decrease in myocardial lesion area and cell death and a reduction in the presence of viral RNA. All DAF-Fc treatment groups had reduced infectious CVB3 recoverable from the heart after infection. DAF-Fc may be a novel therapeutic agent for active myocarditis and acute dilated cardiomyopathy if given early in the infectious period, although more studies are needed to determine its mechanism and efficacy. PMID- 12533689 TI - Inhibition of tumor-induced angiogenesis and matrix-metalloproteinase expression in confrontation cultures of embryoid bodies and tumor spheroids by plant ingredients used in traditional chinese medicine. AB - Tumor-induced angiogenesis is a prerequisite for excessive tumor growth. Blood vessels invade the tumor tissue after degradation of the extracellular matrix scaffold by matrix metalloproteinases (MMPs). Inhibition of MMPs has been therefore suggested to be a useful tool to abolish neoangiogenesis of solid tumors. In the present study, antioxidative plant ingredients used in traditional Chinese medicine were investigated for their capacity to down-regulate MMP expression and to inhibit angiogenesis in embryonic stem cell-derived embryoid bodies and tumor-induced angiogenesis in confrontation cultures consisting of embryoid bodies and multicellular DU-145 prostate tumor spheroids. Embryoid bodies transiently expressed MMP-1, MMP-2, and MMP-9 during the time of differentiation of capillary-like structures. In confrontation cultures, MMP expression was increased compared with control tumor spheroids and embryoid bodies cultivated separately. The increased expression of MMPs in confrontation cultures was a result of elevated levels of reactive oxygen species (ROS) upon confrontation culture and was totally abolished in the presence of the free radical scavenger vitamin E. Incubation of embryoid bodies with baicalein, epicatechin, berberine, and acteoside, which are herbal ingredients used in traditional Chinese medicine, significantly inhibited angiogenesis in embryoid bodies and decreased intracellular ROS levels. Tumor-induced angiogenesis in confrontation cultures was totally abolished in the presence of the free radical scavenger vitamin E. Because herbal ingredients down-regulated MMP expression, we conclude that ROS generated during confrontation culture induce the expression of MMPs that are necessary for endothelial cell invasion into the tumor tissue. PMID- 12533690 TI - Genome-wide analyses on loss of heterozygosity in head and neck squamous cell carcinomas. AB - Head and neck squamous cell carcinoma (HNSCC) is a frequent malignancy with a poor survival rate. Identifying the tumor suppressor gene (TSG) loci by genomic studies is an important step to uncover the molecular mechanisms involved in HNSCC pathogenesis. We therefore performed comprehensive analyses on loss of heterozygosity (LOH) using a genome-wide panel of 191 microsatellite markers in 22 HNSCC samples. We found 53 markers with significantly high LOH (>30%) on 21 chromosomal arms; the highest values of those were observed on 3p, 9p, 13q, 15q, and 17p, corresponding to D3S2432 (67%), D9S921-D9S925 (67%) and GATA62F03 (86%), D13S1493 (60%), D15S211 (62%), and D17S1353 (88%), respectively. Fifteen hot spots of LOH were defined in 13 chromosomal arms: 2q22-23, 4p15.2, 4q24-25, 5q31, 8p23, 9p23-24, 9q31.3, 9q34.2, 10q21, 11q21-22.3, 14q11-13, 14q22.3, 17p13, 18q11, and 19q12 as loci reported previously in HNSCCs. Furthermore, we identified five novel hot spots of LOH on three chromosomal arms in HNSCC at 2q33 (D2S1384), 2q37 (D2S125), 8q12-13 (D8S1136), 8q24 (D8S1128), and 15q21 (D15S211). In conclusion, our comprehensive allelotype analyses have unveiled and confirmed a total of 20 possible TSG loci that could be involved in the development of HNSCC. These results provide useful clues for identification of putative TSGs involved in HNSCC by fine mapping of the suspected regions and subsequent analysis for functional genes. PMID- 12533691 TI - Discordant bone marrow involvement in diffuse large B-cell lymphoma: comparative molecular analysis reveals a heterogeneous group of disorders. AB - Discordant bone marrow (BM) involvement in patients with a diagnosis of large cell non-Hodgkin's lymphoma (NHL) is characterized by marrow infiltrates predominantly composed of small lymphoid cell, cytologically compatible with low grade NHL. Although this phenomenon is well described morphologically, molecular data concerning the relationship of the two lesions are lacking. The aim of the study was to investigate the clonal relationship of discordant lymphoma manifestations by using immunoglobulin heavy chain gene (IgH), as well as bcl-2 rearrangements, as molecular markers. IgH rearrangements were amplified by PCR with consensus primers directed against framework regions 3 or 2 (FR3 and FR2), followed by automated fragment length analysis and sequencing in selected cases. Rearrangements of the bcl-2 gene were identified with primers against the major breakpoint region. Small BM infiltrates were isolated by laser capture microdissection. In addition, immunohistochemistry was performed on paraffin sections using antibodies against CD3, CD10, CD20, bcl-2, bcl-6, p53, and the Ki67 antigen. Paraffin-embedded tissues of 21 cases diagnosed as diffuse large B cell lymphoma (DLBCL) with discordant BM involvement and no previous history of low-grade B-cell NHL were analyzed. After review of immunohistochemical stains, 5 cases were excluded either as concordant BM infiltrates by large-cell lymphoma with abundant reactive T-cells (2 cases) or as benign, reactive lymphoid infiltrates (3 cases), as confirmed by a polyclonal pattern in the IgH analysis. Of the remaining 16 cases, a common clonal origin was confirmed in 8 cases by the presence of an identical clonal IgH rearrangement or bcl-2 rearrangement. In 4 cases, identification of distinct IgH or bcl-2 rearrangements gave evidence for the presence of two clonally unrelated neoplasms. The remaining 4 cases were not evaluable for technical reasons. Morphological, phenotypical, and molecular findings were compatible with a lymphoma of germinal center origin in the majority of cases. However, in 4 cases, flow cytometric analysis of the BM infiltrates revealed a B-cell chronic lymphocytic leukemia phenotype. Two of these cases were clonally related to the DLBCL and thus represented Richter's transformation. In summary, discordant BM infiltrates in DLBCL represent a heterogeneous group of disorders, encompassing cases with a clonally related, clinically occult small-cell component, as well as cases with two clonally distinct, unrelated B-cell neoplasms presenting synchronously at different locations. PMID- 12533692 TI - Immunofluorescence analysis of neutrophil nonmuscle myosin heavy chain-A in MYH9 disorders: association of subcellular localization with MYH9 mutations. AB - The autosomal dominant macrothrombocytopenia with leukocyte inclusions, May Hegglin anomaly, Sebastian syndrome, and Fechtner syndrome, are rare human disorders characterized by a triad of giant platelets, thrombocytopenia, and characteristic Dohle body-like cytoplasmic inclusions in granulocytes. Epstein syndrome is another autosomal dominant macrothrombocytopenia associated with Alport syndrome but without leukocyte inclusions. These disorders are caused by mutations in the same gene, the MYH9, which encodes the nonmuscle myosin heavy chain-A (NMMHCA). The term, MYH9 disorders, has been proposed, but the clinicopathologic basis of MYH9 mutations has been poorly investigated. In this study, a total of 24 cases with MYH9 disorders and suspected cases were subjected to immunofluorescence analysis by a polyclonal antibody against human platelet NMMHCA. Abnormal subcellular localization of NMMHCA was observed in every neutrophil from individuals with MYH9 mutations. Comparison with May-Grunwald Giemsa staining revealed that the NMMHCA always coexisted with the neutrophil inclusion bodies, suggesting that NMMHCA is associated with such bodies. In three cases, neutrophil inclusions were not detected on conventional May-Grunwald Giemsa-stained blood smears but immunofluorescence analysis revealed the abnormal NMMHCA localization. In contrast, cases with Epstein syndrome and the isolated macrothrombocytopenia with normal NMMHCA localization had no MYH9 mutations. An antibody that recognizes the C-terminal 12 mer peptides showed similar immunoreactivity from the patients heterozygous for truncated mutations that abolished the C-terminal epitope, suggesting that normal NMMHCA dimerizes with abnormal NMMHCA to form inclusion bodies. We further propose that the localization pattern can be classified into three groups according to the number, size, and shape of the fluorescence-labeled NMMHCA granule. Immunofluorescence analysis of neutrophil NMMHCA is useful as a screening test for the clear hematopathologic classification of MYH9 disorders. PMID- 12533693 TI - Possible involvement of keratinocyte growth factor and its receptor in enhanced epithelial-cell proliferation and acquired recurrence of middle-ear cholesteatoma. AB - Middle-ear cholesteatoma is characterized by enhanced proliferation of epithelial cells and granular tissue formation. However, the molecular mechanism underlying these pathological changes is largely unknown. Keratinocyte growth factor (KGF) is a mesenchymal cell-derived paracrine growth factor that specifically stimulates epithelial cell proliferation. In the present study, we investigated the possible involvement of KGF and its receptor, KGFR, in the pathogenesis of cholesteatoma using in situ hybridization and immunohistochemistry, respectively. We examined 56 cholesteatoma specimens, and 8 normal skin areas as control. KGF and KGFR expression was examined by immunohistochemistry using rabbit anti-human KGF and anti-human KGFR polyclonal antisera raised in our laboratories against synthetic peptides corresponding to parts of human KGF and KGFR, respectively. KGF protein and mRNA were detected exclusively in stromal fibroblasts and infiltrating T lymphocytes in 80% of cholesteatoma cases, whereas KGFR protein and mRNA were localized in the epithelium in 72% of cases. Assessment of the proliferative activity of cholesteatoma using the labeling index for Ki-67 showed a significantly higher Ki-67 labeling index (66%) in KGF+/KGFR+ cases than other cases. There was a significant correlation between KGF+/KGFR+ expression and recurrence. Our results indicate the possible involvement of both KGF and KGFR in enhanced epithelial cell proliferative activity and recurrence of cholesteatoma. PMID- 12533694 TI - Preservation of tissue RNA in normal saline. PMID- 12533696 TI - Virus complement evasion strategies. AB - The immune system has a variety of tools at its disposal to combat virus infections. These can be subdivided roughly into two categories: 'first line defence', consisting of the non-specific, innate immune system, and 'adaptive immune response', acquired over time following virus infection or vaccination. During evolution, viruses have developed numerous, and often very ingenious, strategies to counteract efficient recognition of virions or virus-infected cells by both innate and adaptive immunity. This review will focus on the different strategies that viruses use to avoid recognition by one of the components of the immune system: the complement system. Complement evasion is of particular importance for viruses, since complement activation is a crucial component of innate immunity (alternative and mannan-binding lectin activation pathway) as well as of adaptive immunity (classical, antibody-dependent complement activation). PMID- 12533697 TI - The human cytomegalovirus genome revisited: comparison with the chimpanzee cytomegalovirus genome. AB - The gene complement of wild-type human cytomegalovirus (HCMV) is incompletely understood, on account of the size and complexity of the viral genome and because laboratory strains have undergone deletions and rearrangements during adaptation to growth in culture. We have determined the sequence (241 087 bp) of chimpanzee cytomegalovirus (CCMV) and have compared it with published HCMV sequences from the laboratory strains AD169 and Toledo, with the aim of clarifying the gene content of wild-type HCMV. The HCMV and CCMV genomes are moderately diverged and essentially collinear. On the basis of conservation of potential protein-coding regions and other sequence features, we have discounted 51 previously proposed HCMV ORFs, modified the interpretations for 24 (including assignments of multiple exons) and proposed ten novel genes. Several errors were detected in the published HCMV sequences. We presently recognize 165 genes in CCMV and 145 in AD169; this compares with an estimate of 189 unique genes for AD169 made in 1990. Our best estimate for the complement of wild-type HCMV is 164 to 167 genes. PMID- 12533698 TI - Human cytomegalovirus UL37 immediate early target minigene RNAs are accurately spliced and polyadenylated. AB - The human cytomegalovirus (HCMV) UL36-38 immediate early (IE) locus encodes proteins required for virus growth. The UL37 IE promoter drives production of differentially spliced and unspliced RNAs. To study their post-transcriptional processing, we generated target minigenes encoding each UL37 RNA splicing substrate. Target 1 RNA, spanning UL37 exon 1 (x1) donor and 2 (x2) acceptor as well as adjacent intronic sequences, but not the UL38 gene, accurately reproduced UL37 x1/x2 RNA splicing in transfected permissive cells. Surprisingly, deletion of distal intronic sequences nt -82 to -143 from the UL37x2 acceptor resulted in aberrant splicing to an upstream non-consensus exonic donor. Target 1 RNAs carry the UL37x1 polyadenylation (PA) signal and site as well as a downstream SV40 early PA signal. Both the UL37x1 and SV40 PA signals are used in wild-type target 1 RNAs but inhibited in UL37x1 PA signal mutants. Alternative RNA splicing of UL37 exons 2 to 3 or 3A as well as exons 3 to 4, observed in HCMV mature UL37 and UL36 spliced RNAs, is accurately reproduced with target minigene RNAs carrying the corresponding UL37 exonic and intronic sequences. Moreover, alternative splicing using two novel UL37 exon 3 consensus splice donors (di and dii) was found in target and in HCMV-infected cell RNA. These results demonstrate that: (i) target minigene RNAs accurately recapitulate the processing of UL37 IE RNAs in the HCMV-infected cell; (ii) precise UL37x1 donor selection is modulated by 3' distal UL37 intronic sequences; and (iii) UL37 exon 3 contains multiple alternative consensus splice donors. PMID- 12533699 TI - Ets-2 Repressor Factor (ERF) mediates repression of the human cytomegalovirus major immediate-early promoter in undifferentiated non-permissive cells. AB - The repression of human cytomegalovirus immediate-early (IE) lytic gene expression is crucial for the maintenance of the latent viral state. By using conditionally permissive cell lines, which provide a good model for the differentiation state-dependent repression of IE gene expression, we have identified several cellular factors that bind to the major immediate-early promoter (MIEP) and whose expression is down-regulated after differentiation to a permissive phenotype. Here we show that the cellular protein Ets-2 Repressor Factor (ERF) physically interacts with the MIEP and represses MIEP activity in undifferentiated non-permissive T2 embryonal carcinoma cells. This factor binds to the dyad element and the 21 bp repeats within the MIEP - regions known to be important for the negative regulation of MIEP activity. Finally, we show that following differentiation to a permissive phenotype ERF's repressive effects are severely abrogated. PMID- 12533700 TI - Human cytomegalovirus immediate-early protein 2 (IE2)-mediated activation of cyclin E is cell-cycle-independent and forces S-phase entry in IE2-arrested cells. AB - In human cytomegalovirus (HCMV) infection, the isolated expression of the viral immediate-early protein 2 (IE2) 86 kDa regulatory protein coincides with an up regulation of cyclin E gene expression, both in fibroblasts and U373 cells. Since IE2 also interferes with cell-cycle progression, it is unclear whether IE2 is a genuine activator of cyclin E or whether IE2-arrested cells contain elevated levels of cyclin E primarily as a consequence of them being arrested at the beginning of S phase. It is important to distinguish between these possibilities in order to define and analyse at a mechanistic level the proliferative and anti proliferative capacities of IE2. Here we have shown that IE2 can activate cyclin E independent of the cell-cycle state and can therefore function as a genuine activator of cyclin E gene expression. A mutant of IE2 that failed to activate cyclin E also failed to promote G1/S transition. Instead, cells became arrested in G1. S-phase entry could be rescued in these cells by co-expression of cyclin E, but these cells still arrested in early S phase, as is the case with wild-type IE2. Our data demonstrate that IE2 can promote two independent cell-cycle functions at the same time: (i) the induction of G1/S transition via up regulation of cyclin E, and (ii) a block in cell-cycle progression in early S phase. In G1, the proliferative activity of IE2 appears to be dominant over the anti-proliferative force, whereas after G1/S transition, this situation is reversed. PMID- 12533701 TI - Synergistic induction of intercellular adhesion molecule-1 by the human cytomegalovirus transactivators IE2p86 and pp71 is mediated via an Sp1-binding site. AB - Human cytomegalovirus (HCMV) infection of transplant recipients is frequently associated with allograft vasculopathy and rejection. One potential mechanism is vascular injury from HCMV-triggered, immunologically mediated processes. HCMV infection has been shown to increase the expression of intercellular adhesion molecule-1 (ICAM-1). The objective of this study was to determine the molecular basis of HCMV-enhanced ICAM-1 gene expression. Transient transfection experiments identified the IE2p86 protein as a potent activator of the ICAM-1 promoter. The tegument protein pp71 showed a strong synergistic effect on IE2p86-mediated ICAM 1 promoter activation. Mutagenesis experiments defined a DNA element from -110 to -42 relative to the transcription start site as responsive for IE2p86. Further point mutations within this DNA element identified an Sp1-binding site that was essential for strong synergistic activation by IE2p86 and pp71. To confirm the activation of ICAM-1 gene expression, human fibroblasts (HFF) as well as endothelial cells (HUVEC) were infected with recombinant IE2p86- and pp71 expressing baculoviruses, respectively. In FACS analysis, a synergistic induction of ICAM-1 was detectable when cells were co-infected with the two recombinant baculoviruses. These findings clearly demonstrate that IE2p86 and pp71 are crucial regulatory factors for HCMV-induced ICAM-1 upregulation. PMID- 12533702 TI - Down-regulation of surface major histocompatibility complex class I by guinea pig cytomegalovirus. AB - Live attenuated strains of human cytomegalovirus are under development as vaccines to prevent birth defects resulting from congenital infections. These strains encode four proteins that inhibit surface expression of MHC class I, presumably to evade cytotoxic T-cell recognition and, perhaps, attenuate induction of immunity. To initiate studies of the role of class I down-regulation on congenital infection and vaccine efficacy, the ability of guinea pig cytomegalovirus to down-regulate class I was examined. Surface class I was specifically down-regulated on infected cells up to 8-fold. Sensitivity to UV irradiation and insensitivity to a viral DNA synthesis inhibitor revealed that immediate early or early viral gene(s) are responsible. Identification of these genes will permit future experiments to evaluate the role of class I down regulation in congenital cytomegalovirus disease and its impact on vaccine efficacy. These findings should be pertinent to understanding human cytomegalovirus disease and may help guide the design of candidate vaccines. PMID- 12533703 TI - Differential detection of B virus and rhesus cytomegalovirus in rhesus macaques. AB - Non-human primate herpesviruses establish and maintain a lifelong persistent infection in immunocompetent hosts in the absence of clinical signs of disease. A fundamental issue for understanding the natural history of non-human primate herpesviruses is whether the viruses are maintained in a truly latent state or one characterized by a low level of chronic expression. To address this issue, a real-time PCR assay was developed to quantify Cercopithecine herpesvirus type 1 (B virus) DNA in mucosal fluids of rhesus macaques. This assay was rapid, sensitive (10 genome copies) and specific for B virus obtained from multiple species of macaques. The shedding profile of B virus was compared to another endemic herpesvirus, rhesus cytomegalovirus (RhCMV), in colony-reared monkeys. Mucosal swabs or saliva samples were taken daily from two groups of seropositive monkeys undergoing either a stressful relocation (group 1) or daily chair restraint (group 2). B virus DNA was detected in mucosal fluids from four animals relocated during the breeding season (group 1) but not from 10 animals moved at other times of the year. No B virus DNA was detected in any group 2 monkey. In contrast, RhCMV DNA was detected in the majority of animals of both groups 1 and 2. Detection of B virus DNA shedding is a relatively rare event associated with the breeding season, while RhCMV DNA is persistently detected in mucosal fluids of most monkeys. PMID- 12533704 TI - Absence of viral antigens on the surface of equine herpesvirus-1-infected peripheral blood mononuclear cells: a strategy to avoid complement-mediated lysis. AB - Equine herpesvirus-1 (EHV-1) may cause abortion in vaccination- and infection immune horses. EHV-1-infected peripheral blood mononuclear cells (PBMCs) play an important role in virus immune evasion. The mechanisms by which infected PBMCs can avoid destruction by EHV-1-specific antibody and equine complement were examined. The majority of EHV-1-infected PBMCs (68.6 %) lacked surface expression of viral antigens and these cells were not susceptible to complement-mediated lysis. In infected PBMCs with surface expression of viral antigens, 63 % showed focal surface expression, whereas 37 % showed general surface expression. General surface expression rendered infected PBMCs susceptible to lysis by antibody and complement (from 5.4 to 31.2 % lysed cells depending on the concentration of antibody and complement). Infected PBMCs with focal surface expression showed significant lysis only in the presence of high concentrations of antibody and complement. Thus, the absence of surface expression protects infected PBMCs against complement-mediated lysis. PMID- 12533705 TI - Transcriptome profile of murine gammaherpesvirus-68 lytic infection. AB - The murine gammaherpesvirus-68 genome encodes 73 protein-coding open reading frames with extensive similarities to human gamma(2) herpesviruses, as well as unique genes and cellular homologues. We performed transcriptome analysis of stage-specific viral RNA during permissive infection using an oligonucleotide based microarray. Using this approach, M4, K3, ORF38, ORF50, ORF57 and ORF73 were designated as immediate-early genes based on cycloheximide treatment. The microarray analysis also identified 10 transcripts with early expression kinetics, 32 transcripts with early-late expression kinetics and 29 transcripts with late expression kinetics. The latter group consisted mainly of structural proteins, and showed high expression levels relative to other viral transcripts. Moreover, we detected all eight tRNA-like transcripts in the presence of cycloheximide and phosphonoacetic acid. Lytic infection with MHV-68 also resulted in a significant reduction in the expression of cellular transcripts included in the DNA chip. This global approach to viral transcript analysis offers a powerful system for examining molecular transitions between lytic and latent virus infections associated with disease pathogenesis. PMID- 12533706 TI - The wood mouse is a natural host for Murid herpesvirus 4. AB - Infection of laboratory mice by the Murid herpesvirus 4 (MHV-4) is a much studied model system for gammaherpesvirus pathogenesis. Little, however, is known about its natural host range, epidemiology and pathogenesis outside the laboratory. We have studied MHV-4 infection in free-living murids in the UK. Using a combination of serology and PCR analysis, we found that MHV-4 was endemic in wood mice (Apodemus sylvaticus) but not in two species of voles (Clethrionomys glareolus, Microtus agrestis). The sites of detection of viral DNA were the lungs and, less commonly, the spleen, emphasizing the importance of the former in virus persistence during natural infection and confirming similar data in laboratory mice. PMID- 12533707 TI - Open reading frame 122 of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus encodes a novel structural protein of occlusion-derived virions. AB - Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HaSNPV) and its closely related variant H. zea SNPV (HzSNPV) contain 20 open reading frames (ORFs) unique among baculoviruses. In this report, the function of HaSNPV ORF 122 (Ha122) is investigated. Ha122 was transcribed as a polyadenylated transcript from 8 h post-infection in infected H. armigera insect cells. 5'RACE analysis indicated that Ha122 transcription starts predominantly in the consensus major late transcription initiation motif DTAAG, around 47 nt upstream of the putative translation start codon, with a minor start at position -89. Using 3'RACE, the transcription stop site mapped 27 nt downstream of the putative translation stop codon. By Western blot analysis using a chicken-derived polyclonal antibody, the product of Ha122 was found in infected cells to be a 21 kDa protein, close to the theoretical size of 21.6 kDa. The Ha122 protein, when fused to green fluorescent protein, was observed in the nuclei of H. armigera cells but only in conjunction with wild-type HaSNPV infection. The 21 kDa protein was located specifically in the nucleocapsid of occlusion-derived virions (ODVs) and not in that of budded virus. The available data suggest that Ha122 is a functional ORF of HaSNPV and HzSNPV and that the 21 kDa protein is a novel specific component of baculovirus ODVs. PMID- 12533708 TI - Characterization of Mamestra configurata nucleopolyhedrovirus enhancin and its functional analysis via expression in an Autographa californica M nucleopolyhedrovirus recombinant. AB - Enhancin genes have been identified in a number of baculoviruses and enhancin proteins are characterized by their ability to enhance the oral infectivity of heterologous baculoviruses in various lepidopteran insects. Here, we describe the putative enhancin gene from Mamestra configurata nucleopolyhedrovirus (MacoNPV), only the second NPV in which an enhancin-like ORF has been identified. The putative enhancin gene from MacoNPV has a typical baculovirus late promoter (ATAAG) 15 bp upstream from the ATG codon. The enhancin ORF encodes an 847 amino acid protein with a predicted molecular mass of 98 kDa and contains a conserved zinc-binding domain (HEIAH) common to metalloproteases. The MacoNPV enhancin shows approximately 20 % amino acid identity with other baculovirus enhancins. An Autographa californica M nucleopolyhedrovirus (AcMNPV) recombinant, AcMNPV-enMP2, expressing the MacoNPV enhancin gene under control of its native promoter was developed and characterized. Northern blot analysis showed expression of enhancin from 24 through 72 h post-infection. In 2nd-instar Trichoplusia ni larvae, the LD50 of the AcMNPV-enMP2 recombinant was 2.8 polyhedral inclusion bodies (PIB) per larva, 4.4 times lower than that of AcMNPV E2 wild-type virus (12.4 PIB per larva). At biologically equivalent doses, i.e. LD90, the survival time 50 % (ST50) of AcMNPV-enMP2 recombinant and AcMNPV E2 wild-type viruses were not significantly different. PMID- 12533709 TI - Adeno-associated virus integrates site-specifically into human chromosome 19 in either orientation and with equal kinetics and frequency. AB - Adeno-associated virus type 2 (AAV-2) establishes latency by site-specific integration into a unique locus, AAVS1, on human chromosome 19 (chr19). To study the kinetics and frequency of chr19-specific integration, a rapid, sensitive and quantitative real-time PCR assay specific for AAV inverted terminal repeat (ITR) chr19 junction sequences was developed. Since the assay only detected right-hand AAV ITR-specific integration events, the development of a complementary left-hand ITR-specific real-time PCR assay is described. The time-course of left-hand ITR dependent AAV integration at AAVS1 of chr19 was determined in AAV-2-infected HeLa cells. Both the kinetics and frequencies of left-hand ITR-dependent integration were found to be similar to those of the right-hand ITR. In addition, left-hand ITR-specific fusion sequences and chromosomal breakpoints within AAVS1 were variable, yet were the same as those found in right-hand ITR-chr19 junction sequences. Thus, the AAV-2 genome integrates site-specifically into chr19 with similar efficiency in either orientation. PMID- 12533710 TI - Priming of cytotoxic T cell responses to exogenous hepatitis B virus core antigen is B cell dependent. AB - The hepatitis B virus (HBV) core antigen (HBcAg) has a unique ability to bind a high frequency of naive human and murine B cells. The role of HBcAg-binding naive B cells in the immunogenicity of HBcAg is not clear. The HBcAg-binding properties of naive B cells were characterized using HBcAg particles with mutated spike region (residues 76-85) sequences. Deletion of residues 76-85 (HBcDelta76-85) destroyed naive B cell binding, whereas deletion of residues 79-85 did not. HBcAg particles with an Ile instead of the natural Ala at position 80 did not bind naive B cells, whereas reversion of Ile80-->Ala restored B cell binding. Destroying the B cell-binding ability of HBcAg had a marginal effect on the overall B cell immunogenicity of the different particles, suggesting that they were equally efficient in priming T helper cells. Therefore, the importance of HBcAg-binding B cells is studied with relation to the priming of HBcAg-specific cytotoxic T cells (CTLs). The role of HBcAg-binding B cells in the priming of HBcAg-specific CTLs was evaluated by immunization with endogenous HBcAg (DNA immunization) and exogenous recombinant HBcAg particles. Endogenous HBcAg primed HBcAg-specific CTLs in wild-type and B cell-deficient mice, whereas exogenous HBcAg primed HBcAg-specific CTLs only in wild-type mice. Importantly, HBcDelta76 85 did not prime CTLs despite the presence of B cells. Thus, the ability of exogenous HBcAg particles to prime specific CTLs is B cell dependent, suggesting a possible role for HBcAg-binding B cells in HBV infections. PMID- 12533711 TI - Molecular epidemiology of gibbon hepatitis B virus transmission. AB - Although transmission of human hepatitis B virus (HBV) variants to nonhuman primates is well documented, it remains to be elucidated whether nonhuman primate HBV is transmissible to humans. The prevalence and transmission routes of gibbon HBV were analysed in 101 captive gibbons in Thailand. Approximately 40 % of these animals showed at least one marker of HBV infection; 19 animals were chronic HBV carriers, characterized by elevated levels of alanine amino transferase and the presence of HBV DNA. Some of the chronic animals were found to be anti-HBc (HBV core antigen) negative (4 of 19), while precore promoter point mutations (nt 1762 or 1764) were determined in four animals by RFLP analysis. Phylogenetic tree analysis of the complete surface gene sequences revealed that gibbon viruses clustered separately from hepadnaviruses of other hosts. Evidence for horizontal and vertical transmission in captive gibbons was obtained. HBV DNA was also detected in the saliva of HBV carrier gibbons. Although some of the animal caretakers at the Krabok Koo Wildlife Breeding Centre were found to be chronic HBV carriers, genotype and sequence analysis did not reveal any evidence for zoonotic disease transmission. PMID- 12533712 TI - Positively charged sequences of human papillomavirus type 16 capsid proteins are sufficient to mediate gene transfer into target cells via the heparan sulfate receptor. AB - Using synthetic peptides we have shown that positively charged sequences present at the C terminus of the L1 protein and the N and C termini of the L2 protein of human papillomavirus type 16 (HPV-16) bind to both DNA and heparan sulfate receptors. Moreover, these short amino acid sequences are sufficient to mediate gene transfer in COS-7 cells. The L1 proteins of other HPVs were shown to contain one or two DNA- and heparin-binding sequences that have the capacity to transfer genes. These DNA-binding sequences also recognized the enhancing packaging sequence of bovine papillomavirus type 1. The results suggest that the L2 protein could participate in DNA packaging during maturation of virions. PMID- 12533713 TI - A new virus infecting Myzus persicae has a genome organization similar to the species of the genus Densovirus. AB - The genomic sequence of a new icosahedral DNA virus infecting Myzus persicae has been determined. Analysis of 5499 nt of the viral genome revealed five open reading frames (ORFs) evenly distributed in the 5' half of both DNA strands. Three ORFs (ORF1-3) share the same strand, while two other ORFs (ORF4 and ORF5) are detected in the complementary sequence. The overall genomic organization is similar to that of species from the genus DENSOVIRUS: ORFs 1-3 most likely encode the non-structural proteins, since their putative products contain conserved replication motifs, NTP-binding domains and helicase domains similar to those found in the NS-1 protein of parvoviruses. The deduced amino acid sequences from ORFs 4 and 5 show sequence similarities with the structural proteins of the members of the genus DENSOVIRUS: These data indicate that this virus is a new species of the genus Densovirus in the family PARVOVIRIDAE: The virus was tentatively named Myzus persicae densovirus. PMID- 12533714 TI - Entry of alphaviruses at the plasma membrane converts the viral surface proteins into an ion-permeable pore that can be detected by electrophysiological analyses of whole-cell membrane currents. AB - Alphaviruses are small enveloped viruses that have been used extensively as model enveloped viruses. During infection, virus particles are taken up into endosomes, where a low pH activates the viral fusion protein, E1. Fusion of the viral and the endosomal membranes releases the viral core into the cytoplasm where cores are disassembled by interaction with 60S ribosomal subunits. Recently, we have shown that in vitro this disassembly is strongly stimulated by low pH. We have proposed that after entry of the core into the cytoplasm, the viral membrane proteins that have been transferred to the endosomal membrane form an ion permeable pore in the endosome. The resulting flow of protons from the endosome into the cytoplasm through this pore could generate a low-pH environment for core disassembly in vivo. Here we report two types of analysis aimed at the identification of such pores. First, the release of [3H]choline from the interior of liposomes was analysed in the presence of virus particles and viral proteins. Secondly, cells were infected with Sindbis or Semliki Forest alphaviruses at the plasma membrane and the possible generation of ion-permeable pores during this process was analysed by whole-cell voltage clamp analysis of the membrane current. The results obtained indicated that the proposed pores are in fact generated and allowed us to identify the formation of individual pores. Available evidence indicates that the alphavirus E1 protein probably forms these pores. Proteins homologous to the alphavirus E1 protein are present in flaviviruses and hepatitis C virus. PMID- 12533715 TI - Cleavage of protein prM is necessary for infection of BHK-21 cells by tick-borne encephalitis virus. AB - Flavivirus particles are synthesized in an immature form containing heterodimers of the proteins prM and E. Shortly before release from the cell, prM is cleaved by the host protease furin to yield mature virions. In this study, the furin mediated cleavage of the tick-borne encephalitis (TBE) virus protein prM was prevented by specific mutagenesis of the cleavage site. This resulted in the production of immature TBE virions, which were shown to be completely non infectious in BHK-21 cells. This finding contrasted with previous studies in which immature flavivirus particles produced by other techniques were shown to have considerable residual infectivity. The structural integrity of the mutant virus particles was confirmed by the characterization of physical and antigenic properties. Most importantly, infectivity could be restored by the addition of trypsin, which presumably cleaved protein prM at one of the monobasic sites retained in the mutated sequence. In the presence of trypsin, the mutant could be passaged repeatedly in BHK-21 cells, but if the protease was removed, the activated particles could initiate only a single round of infection, which again generated non-infectious virus progeny. These observations provide evidence that the infectivity of flaviviruses depends on the endoproteolytic cleavage of protein prM, which probably has a regulatory function rather than a direct role in virus entry. Moreover, the results illustrate that mutation of the furin cleavage site is a convenient way to produce single-round infectious flavivirus particles, which may be useful in vaccine and vector development. PMID- 12533716 TI - Virus-cell interactions in the induction of type 1 interferon by influenza virus in mouse spleen cells. AB - Inactivated influenza A virus and fixed, virus-infected cells induce type 1 interferon (IFN-alpha/beta) production in murine splenocytes. In this study, we have explored the nature of the virus-spleen cell interaction that leads to IFN alpha/beta induction and the reason for the poor response to some virus strains. IFN-alpha/beta induction by horse serum-sensitive, but not -resistant, strains of influenza virus was inhibited in the presence of horse serum, indicating that binding of the virus to sialylated cell receptors is a necessary step in the induction process. Furthermore, influenza viruses A/PR/8/34 (H1N1) and A/WS/33 (H1N1), which were poor inducers of IFN-alpha/beta in spleen cells, were shown to have a more active neuraminidase than strains that induced higher IFN levels, and IFN-alpha/beta induction by A/PR/8/34 (H1N1) and A/WS/33 (H1N1) was restored in the presence of a neuraminidase inhibitor. Growth of virus in different cell types altered the level of IFN-alpha/beta induced in spleen cells by particular virus strains, suggesting that the nature of the carbohydrate moieties on the viral glycoproteins may also influence IFN-alpha/beta induction in this system. Consistent with this notion, treatment of egg-grown virus with periodate to oxidize viral carbohydrate greatly reduced its capacity for IFN-alpha/beta induction. Furthermore, induction of IFN-alpha/beta was inhibited in the presence of the saccharides yeast mannan and laminarin. Together these findings indicate: (i) a requirement for interaction of the virus with sialylated receptors on the IFN-producing cell; (ii) an influence of viral carbohydrate on the response; and (iii) possible involvement of a lectin-like receptor on the IFN-producing cell in the induction of IFN-alpha/beta or in regulation of this response. PMID- 12533717 TI - Immunogenicity in Mamu-A*01 rhesus macaques of a CCR5-tropic human immunodeficiency virus type 1 envelope from the primary isolate (Bx08) after synthetic DNA prime and recombinant adenovirus 5 boost. AB - Envelopes of primary R5-tropic human immunodeficiency virus type 1 (HIV-1) isolates may be particularly relevant for vaccine purposes and should be evaluated for immunogenicity in animals including macaques before carrying out human vaccine trials. In the present study, the immunogenicities of synthetic HIV 1 env DNA vaccines, which had been derived from the early primary isolate Bx08 and contain humanized codons, were evaluated in mice, guinea pigs and rhesus macaques. Neutralization sensitivity of the HIV-1(Bx08) isolate was found to resemble that of other primary isolate prototypes. Immunogenicity of gp120 delivered as codon-optimized DNA vaccine was comparable to that of recombinant gp120 protein plus adjuvant in mice. Similarly, DNA vaccination of guinea pigs with synthetic gp140(Bx08) and gp150(Bx08) DNA induced a strong antibody response independent of the gene construct and DNA immunization route. Mamu-A*01 rhesus macaques were DNA vaccinated with synthetic gp150(Bx08) or gp140(Bx08) DNA and boosted with a replication-deficient recombinant human adenovirus type 5 expressing a synthetic gp120(Bx08) gene. DNA-vaccinated rhesus macaques developed specific CD8+ T lymphocyte responses and anti-rgp120(IIIb) antibody responses. Both the humoral and cellular responses were significantly improved following intramuscular boosting with the recombinant adenovirus. The demonstrated humoral and cellular immunogenicities of these HIV Bx08 Env vaccines in non-human primates encourages their further development as one component in candidate HIV vaccines for humans. PMID- 12533718 TI - Rabbit endogenous retrovirus-H encodes a functional protease. AB - Recent studies have revealed that 'human retrovirus-5' sequences found in human samples belong to a rabbit endogenous retrovirus family named RERV-H. A part of the gag-pro region of the RERV-H genome was amplified by PCR from DNA in human samples and several forms of RERV-H protease were expressed in bacteria. The RERV H protease was able to cleave itself from a precursor protein and was also able to cleave the RERV-H Gag polyprotein precursor in vitro whereas a form of the protease with a mutation engineered into the active site was inactive. Potential N- and C-terminal autocleavage sites were characterized. The RERV-H protease was sensitive to pepstatin A, showing it to be an aspartic protease. Moreover, it was strongly inhibited by PYVPheStaAMT, a pseudopeptide inhibitor specific for Mason Pfizer monkey virus and avian myeloblastosis-associated virus. A structural model of the RERV-H protease was constructed that, together with the activity data, confirms that this is a retroviral aspartic protease. PMID- 12533719 TI - A possible overwintering mechanism for bluetongue virus in the absence of the insect vector. AB - Bluetongue virus (BTV) and several other Orbivirus species are transmitted between mammalian hosts via bites from adults of certain species of Culicoides midges. However, BTV can survive for 9-12 months (typically during the winter), in the absence of adult vectors, with no detectable cases of viraemia, disease or seroconversion in the host. The survival of the virus from one 'vector season' to the next is called 'overwintering' but the mechanism involved is not fully understood. It is demonstrated that BTV can persistently infect ovine gammadelta T-cells in vitro, a process that may also occur during infection and viraemia in mammalian hosts, thus providing a mechanism for virus persistence. Interaction of persistently BTV-infected gammadelta T-cells with antibody to the gammadelta T cell-specific surface molecule WC-1 resulted in conversion to a lytic infection and increased virus release. Skin fibroblasts induce a similar conversion, indicating that they express a counter ligand for WC-1. Feeding of Culicoides midges induces skin inflammation, which is accompanied by recruitment of large numbers of activated gammadelta T-cells. The interaction of persistently infected gammadelta T-cells with skin fibroblasts would result in increased virus production at 'biting sites', favouring transmission to the insect vector. This suggested mechanism might also involve up-regulation of the WC-1 ligand at inflamed sites. It has been shown previously that cleavage of virus surface proteins by protease enzymes (which may also be associated with inflammation) generates infectious subvirus particles that have enhanced infectivity (100 times) for the insect vector. PMID- 12533721 TI - Genetic diversity and biological variation among California isolates of Cucumber mosaic virus. AB - Genetic diversity and biological variation were compared for California isolates of Cucumber mosaic virus (CMV). These fell into five pathotypes based on their reactions on three cucurbits including a susceptible squash, a melon with conventional resistance and a commercial CMV-resistant transgenic squash. Thirty three isolates infected and caused symptoms on CMV-resistant transgenic squash. Forty-two isolates infected the CMV-resistant melon, but only 25 isolates infected both. Single-strand conformation polymorphism (SSCP) analysis was used to differentiate 81 California isolates into 14 groups, and the coat protein (CP) genes of 27 isolates with distinct and indistinguishable SSCP patterns were sequenced. Fourteen isolates corresponding to the different SSCP patterns were also used for phylogenetic analysis. Seventy-nine isolates belonged to CMV subgroup IA, but two belonged to CMV subgroup IB. This is the first report of subgroup IB isolates in the Americas. All CMV isolates had a nucleotide identity greater than or equal to 93.24 %. There was no correlation between CP gene variation and geographical origin, collection year, original host plant, or between the degree of CP amino acid sequence identity and the capacity to overcome transgenic and/or conventional resistance. SSCP and sequence analyses were used to compare 33 CMV isolates on CMV-resistant transgenic squash and susceptible pumpkin plants. One isolate showed sequence differences between these two hosts, but this was not due to recombination or selection pressure of transgenic resistance. CMV isolates capable of infecting cucurbits with conventional and transgenic CMV resistance were present in California, even before CMV transgenic material was available. PMID- 12533720 TI - Characterization of DNAbeta associated with begomoviruses in China and evidence for co-evolution with their cognate viral DNA-A. AB - Eighteen samples of begomoviruses isolated from tobacco, tomato and weed species in Yunnan, China were found to be associated with DNAbeta molecules, for which the complete nucleotide sequences were found to contain 1333-1355 nt. The 18 DNAbeta molecules identified consist of three main types, each associated with a different begomovirus species: 72-99 % nucleotide identity was found within one type, but only 39-57 % identity was found between types. All the DNAbeta molecules reported here and elsewhere contain a 115 nt conserved region that has 93-100 % identity with a consensus sequence, and have a common ORF encoding 118 amino acids on the complementary strand (designated C1). Co-agroinoculation of the DNA-A component of Tomato yellow leaf curl China virus tobacco isolate Y10, with its associated DNAbeta (Y10beta), shows this DNAbeta to be involved in symptom induction in tobacco and tomato. The in-frame ATG mutation of C1 of Y10beta caused much milder symptoms as compared with wild Y10beta, indicating a functional role for this ORF. Pairwise nucleotide sequence identity comparisons of DNAbeta molecules and their cognate viral DNA-A molecules indicate that DNAbeta molecules have co-evolved with their cognate helper viruses. Recombination between DNAbeta molecules is documented and a DNAbeta species concept is proposed and discussed. PMID- 12533722 TI - Changes of retinal glutamate transporter GLT-1 mRNA levels following optic nerve damage. AB - PURPOSE: Under physiological conditions, levels of the excitatory neurotransmitter glutamate within the retina are regulated by retinal glutamate transporters to prevent toxic accumulation. Alterations in this glutamate buffering have been implicated in retinal ganglion cell (RGC) death. We quantified the changes in the level of glutamate transporter mRNA in a model of acute rat optic nerve injury. METHODS: Optic nerve damage was induced in one eye of 25 adult Wistar rats by partial optic nerve crush (ONC). Total mRNA levels of the retinal glutamate transporter GLT-1 (EAAT-2) were determined by quantitative real-time PCR. GLT-1 mRNA levels were measured 1, 3, 7, 14, and 28 days following optic nerve injury. Additionally, control values were obtained from the retinas of five control rats (sham-crush). RESULTS: In the very early phase (1 day post ONC), a 3.9 fold increase in levels of GLT-1 mRNA was observed in the ONC retinae compared with control eyes. This was followed by a rapid decrease towards control levels at day 3 post-ONC. GLT-1 mRNA levels remained up to 14 days post-crush. However, in the late phase post-ONC (day 28), the level of GLT-1 mRNA increased again, but still remained not significant to control levels. CONCLUSIONS: Changes in GLT-1 mRNA expression following axonal trauma of RGCs can lead to an imbalance of glutamate homeostasis. This may cause local accumulation of toxic concentrations of the neurotransmitter glutamate and further irreversible excitotoxic damage of RGCs. PMID- 12533724 TI - On-line opportunities with the Journal of Pediatric Health Care. PMID- 12533723 TI - The UV responsive elements in the human mimecan promoter: a functional characterization. AB - PURPOSE: A major environmental stress encountered by humans is solar UV light, which can cause a spectrum of eye diseases, such as photokeratitis, cataract, pterygia, and ocular neoplasms. Mammalian defense mechanisms in response to adverse effects of UV light result in induction of a number of genes. Studies on the transcriptional regulation of genes that are expressed in the eye will increase understanding of both the physiological functions of these genes in the mammalian UV response, and the molecular bases for abnormalities associated with the above diseases. Mimecan is an extracellular matrix proteoglycan that is abundantly expressed in the cornea. The purpose of this study was to determine and characterize the UV responsive regulatory elements of the human mimecan promoter. METHODS: Transcriptional activity of the promoter was evaluated, before and after UV irradiation, using transient transfection of human mimecan promoter/luciferase reporter constructs into corneal keratocytes and non-corneal cells. Site directed mutagenesis and corresponding functional assays were used to determine the contribution of UV responsive regions to human mimecan transcription. Co-transfection experiments were used to investigate the role of transcription factors that bind these elements in the promoter and mediate the UV response. mRNA expression was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The shortest promoter construct that was strongly activated following UV irradiation contained three initiator elements, an E-box element that is conserved between species, and the entire first intron of the human mimecan gene. Deletion of the intronic p53 binding site from this construct considerably diminished transcription and the UV response of the promoter. Surprisingly, deletion of the E-box sequence from this construct completely abolished both transcription and UV response of the promoter. These results demonstrated that the E-box is essential to transcription of the human mimecan gene and also is required for activation by p53. The role of the E-box, and the E box binding protein, USF-1, in transcription and UV responses of the human mimecan promoter were confirmed by co-transfection experiments using dominant negative transcription factor, A-USF. In addition to these positive regulators, we demonstrate that the region between nucleotides -1314 and -1907 contains a transcriptional repressor site that is active in a time dependent manner following UV irradiation. Finally, we show that UV irradiation results in changes in mimecan mRNA levels in bovine corneal keratocytes in a time-dependent manner. CONCLUSIONS: The human mimecan promoter contains several UV responsive regulatory elements that are conserved between human and bovine species and include the intronic p53 DNA binding site, the E-box in the proximal promoter, and the region between nucleotides -1314 and -1907. The E-box plays an important role in transcription and UV response of the human mimecan promoter. UV irradiation modulates expression of mimecan mRNA in bovine corneal keratocytes and non corneal cells. PMID- 12533725 TI - Using a matrix as an educational approach to asthma. AB - Health care providers agree that asthma care must be individualized to fit the need of the child and family. A written plan in the form of a matrix enables the health care provider to give families a step-by-step individualized plan of care for asthma. This article discusses the use of a long-term treatment plan in matrix form as an educational tool for health care providers and for families. The written long-term plan can be used to guide treatment for any level of asthma severity. Following a review of asthma pathogenesis and diagnosis, categories of medications are presented, along with their role in a long-term treatment plan. The National Heart, Lung, and Blood Institute guidelines are used as a basis for the recommendations in the long-term treatment plan. PMID- 12533726 TI - The obese child: motivation as a tool for exercise. AB - The purpose of this article is to describe the importance of motivation in enhancing the participation of obese children in exercise activities. Recognizing the different influence that are important determinants of exercise behavior in children may help pediatric nurse practitioners play a significant role in advising parents and others of the need to offer positive, constructive, and immediate feedback without being evaluative, critical, or demanding. By addressing the problems of obesity and low fitness levels early in the child's life, a significant step can be taken toward reversing the negative trends of this unhealthy and potentially dangerous condition. PMID- 12533727 TI - President's message: Celebrating the pearl that is NAPNAP. PMID- 12533728 TI - The Kids Cafe: a program to reduce child hunger. AB - Hunger and food insecurity affect nearly 12 million children in the United States. Poverty is the foremost reason for hunger and food insecurity, but even the working poor sometimes have difficulty providing enough food for their household. Undernourished children may not present with severe clinical symptoms, but their ability to learn and psychosocial behavior can be affected. Feeding programs such as Kids Cafe can help decrease child hunger while improving learning and overall health. Kids Cafe's are operated by local food banks and sponsored by America's Second Harvest in partnership with ConAgra Foods, Inc. This article reviews the problem of child hunger and describes the Kids Cafe Program. PMID- 12533729 TI - Conflict and conflict resolution: parent and young teen perceptions. AB - INTRODUCTION: The purpose of this preliminary study was to describe a novel approach to examine the thinking of parents and young teens about conflict and conflict resolution in their relationship. The novel approach was that teens and parents were asked to solve, in focus groups, a hypothetical conflict situation without, and then with, a structured conflict resolution guide. METHODS: Two focus groups were conducted. The resulting data were analyzed with use of content analysis from a sample of 8 parent-young teen dyads. The young teens and their parents participated separately from one another in the focus groups. RESULTS: Data demonstrated that the young teens (a) thought parents or siblings initiated most disagreements and that such disagreements were routine, and (b) handled conflict with their parents by trying to prevent it. If a conflict ensued, they used emotion, aggression, cooling off, accepting some of the blame, or submission to resolve it. Parents viewed the disagreements as (a) representing their struggles with their role as a parent or (b) opportunities to instill a sense of intrinsic responsibility in their child. Parents used the strategies of setting clear expectations, parental authority, negotiation, cooling down, and feedback to solve disagreements with their teenage children. DISCUSSION: We concluded that these parents and young teens do not use a systematic method of solving disagreements but that with structured guidance, the parents and teens were able to resolve conflicts. PMID- 12533730 TI - The pediatric health care home. PMID- 12533731 TI - Why are some children still uninsured? AB - INTRODUCTION: Nationally, 24% of low-income children remain uninsured after the implementation of the State Children's Health Insurance Program (SCHIP). METHOD: The purpose of this study was to understand why children remain uninsured by comparing children with insurance to those without it. Using a cross-sectional survey design, 392 low-income parents were interviewed. RESULTS: There were distinct profiles for the privately insured, Medicaid-insured and uninsured groups. Statistically significant differences were found across the three groups in income, working status of the adults, education, health status of the adult and child, and in the utilization of health care. Parents of the uninsured children were less knowledgeable about the application process. DISCUSSION: Parents of uninsured children face multiple life challenges that may interfere with the enrollment process. Health problems, work schedules, and lack of knowledge may all need to be addressed before we can decrease the number of uninsured children in our nation. PMID- 12533732 TI - Pediatric orthopedic physical examination of the infant: a 5-minute assessment. PMID- 12533733 TI - Evaluation and management of scoliosis. PMID- 12533734 TI - It's a nail biter! PMID- 12533735 TI - Evidence-based practice: the paradigm shift. PMID- 12533738 TI - Chemotherapy of multiple myeloma: melphalan--40 years old and still going strong. PMID- 12533739 TI - The role of cytotoxic therapy with hematopoietic stem cell transplantation in the therapy of multiple myeloma: an evidence-based review. AB - Evidence supporting the role of hematopoietic stem cell transplantation (SCT) in the therapy of multiple myeloma (MM) is presented and critically evaluated in this systematic evidence-based review. Specific criteria were used for searching the published medical literature and for grading the quality of the evidence, the strength of the evidence, and the strength of the treatment recommendations. Treatment recommendations based on the evidence presented in the review were made unanimously by a panel of MM experts. Recommendations for SCT as an effective therapy for MM include the following: SCT is preferred to standard chemotherapy as de novo therapy; SCT is preferred as de novo rather than salvage therapy; autologous peripheral blood stem cell transplantation (PBSCT) is preferred to bone marrow transplantation (BMT); and melphalan is preferred to melphalan plus total body irradiation as the conditioning regimen for autologous SCT. Recommendations that SCT is not effective include the following: current purging techniques of bone marrow. Recommendations of equivalence include the following: PBSCT using CD34+ selected or unselected stem cells. No recommendation is made for indications or transplantation techniques that have not been adequately studied, including the following: SCT versus standard chemotherapy as salvage therapy, tandem autologous SCT, autologous or allogeneic SCT as a high-dose sequential regimen, allogeneic BMT versus PBSCT, a preferred allogeneic myeloablative or non-myeloablative conditioning regimen, and maintenance therapy post-autologous SCT with interferon alpha post-SCT. The priority area of needed future research is maintenance therapy posttransplantation with nothing versus interferon alpha versus other agents such as corticosteroids or thalidomide or its derivatives. PMID- 12533741 TI - Gastrointestinal involvement in chronic graft-versus-host disease: a clinicopathologic study. AB - The original histopathologic description of chronic graft-versus-host disease (CGVHD) of the gastrointestinal (GI) tract was from autopsy series. There is little information on the evaluation of living patients with CGVHD and GI symptoms. We reviewed data on 40 consecutive patients with CGVHD and persistent GI symptoms who underwent endoscopic examinations. The diagnosis of CGVHD in these 40 patients was made on the basis of clinical criteria and confirmed by histology of other involved organs in 70%. Patients had progressive (in 19 patients, or 48%), quiescent (in 11, or 27%) or de novo-type (in 10, or 25%) onset of their CGVHD. Four groups were defined based on the following histologic criteria: (1) consistent with acute GI GVHD if there was marked apoptosis with or without cryptitis, (2) suggestive of acute GI GVHD if there was scattered apoptosis with or without cryptitis, (3) suggestive of chronic GI GVHD if there were at least 2 histologic indicators of chronicity such as fibrosis and significant crypt distortion, and (4) no histologic evidence of GVHD. Results of microbiologic, radiologic, and malabsorption studies, if performed, were also retrieved. Median time from diagnosis of CGVHD to GI endoscopy was 4.5 months (0 109 months). The major GI symptoms at the time of endoscopy were diarrhea, abdominal pain/cramping, nausea/vomiting, weight loss, dysphagia, and early satiety. The endoscopic examination was nonspecific for the diagnosis of GI GVHD except for diffuse mucosal sloughing. Based on the histologic criteria in 22 patients with biopsies, 13 cases (59%) were considered to have acute GI GVHD, and 3 cases (14%) were felt to show possible chronic GI GVHD; changes of both acute and chronic GVHD were seen in 6 (27%) cases. GI dysmotility was diagnosed in 7 (18%) patients, including 2 of the patients who had histologic changes suggestive of chronic GVHD. Other causes of the GI symptoms included infection, drug side effect, and malabsorption. In conclusion, GI involvement by acute GVHD appears to be a major cause of persistent GI symptoms in patients with chronic GVHD. An isolated form of chronic GI GVHD confirmed by histology is an uncommon phenomenon in the actual clinical setting. PMID- 12533740 TI - Chronic graft-versus-host disease: a prospective cohort study. AB - Chronic graft-versus-host disease (CGVHD) is a major cause of morbidity and mortality following allogeneic bone marrow transplantation (BMT). We studied 159 patients with CGVHD longitudinally to characterize the natural history of CGVHD and identify reliable predictors of response and long-term mortality. Rates of response to treatment were 61%, 53%, and 50% at 6 months, 1 year, and 2 years, respectively. A high incidence of infections (7 of 1,000 patient-days at 0 to 6 months, 2.5 of 1,000 patient-days at 6 months to 1 year, and 0.6 of 1,000 patient days at 1 to 2 years) was observed. After a median follow-up of 8.4 years, an overall survival rate of 40% was observed. The overall survival rate was 63% (95% confidence interval [CI], 56%-71%) at 1 year, 51% (95% CI, 43%-59%) at 2 years, and 39% (95% CI, 31%-47%) at 10 years. In multivariate analysis, age older than 20 years (RR = 1.5; 95% CI, 0.9%-2.5%; P =.09), progressive onset of CGVHD (RR = 1.6; 95% CI, 1.0%-2.4%; P =.04), platelet count of <100,000/ microL (RR = 2.1; 95% CI, 1.3%-3.4%; P =.001), and GI involvement (RR = 1.5; 95% CI, 1.0%-2.4%; P =.05) were associated with increased mortality. Among patients surviving more than 6 months, no response (RR = 4.5; 95% CI, 1.9%-10.5%; P =.0006) and partial response (RR = 2.5; 95% CI, 1.1%-6.1%; P =.04) to treatment at 6 months also were significant predictors of mortality. The prevalence of active CGVHD was 33% at 2 years. However, the cumulative incidence of successful discontinuation of therapy was only 13% at 2 years. Among patients with clinical resolution of CGVHD, only 18% were off immunosuppressive therapy by 2 years, and 89% by 4 years. Despite high initial response rates, a large majority of patients had active disease requiring prolonged immunosuppression. This requires improved infection prevention for a longer time. Recognition of a high-risk group should facilitate assignment of more intensified regimens. Better treatment regimens need to be identified to improve survival and limit toxicity of prolonged immunosuppression. PMID- 12533742 TI - A comparison of related donor peripheral blood and bone marrow transplants: importance of late-onset chronic graft-versus-host disease and infections. AB - Peripheral blood hematopoietic stem cell (PBSC) transplants have been shown to result in more rapid engraftment than standard bone marrow transplants (BMTs). Little comparative data exist regarding complications in patients receiving transplants using these stem cell sources. In our study, 97 adults with advanced hematologic malignancies who received allogeneic PBSC transplants were compared with 97 adults who received allogeneic BMTs using identical preparative regimens and support parameters. The incidence of systemic infections and other major complications occurring within the first year after transplantation were calculated in both groups. Proportional hazard analysis was used to examine risk factors for death and complications in both groups. Patients receiving PBSC transplants had more rapid neutrophil (17 days versus 24 days; P <.001) and platelet engraftment (28 days versus 47 days; P <.001) than BMT recipients. The survival rate at 2 years was 38% in PBSC transplant recipients and 28% in marrow recipients (P =.08). There was no difference in rates of grade II to IV acute graft-versus-host disease (GVHD) between groups (PBSC 46%, BMT 51%; P =.3). PBSC transplant recipients were more likely to develop chronic GVHD after 180 days (hazard ratio 2.2; P =.05). Accompanying this "late-onset chronic GVHD," a pattern of more frequent late systemic fungal and cytomegalovirus infections was observed in PBSC transplant recipients. In conclusion, although PBSC transplant recipients engraft more quickly than BMT recipients and have somewhat better 2 year survival rates, they develop more frequent late-onset chronic GVHD and may have more late fungal and cytomegalovirus infections than marrow recipients. Further studies must examine this late-onset chronic GVHD and better characterize immune reconstitution in PBSC transplant recipients to understand their effects on patient recovery. PMID- 12533744 TI - NTP toxicology and carcinogensis studies of vanadium pentoxide (CAS No. 1314-62 1) in F344/N rats and B6C3F1 mice (inhalation). AB - Vanadium pentoxide, commercially the most important compound of vanadium, presents a potential occupational hazard during the cleaning of oil-fired boilers and furnaces, the handling of catalysts, and during the refining, processing, or burning of vanadium-rich mineral ores or fossil fuels. Vanadium pentoxide was nominated for study by the National Cancer Institute as a representative of the metals class study. Male and female F344/N rats and B6C3F1 mice were exposed to vanadium pentoxide (99% pure) by inhalation for 16 days, 14 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium and mouse peripheral blood. 16-DAY STUDY IN RATS: Groups of five male and five female rats were exposed to particulate aerosols of vanadium pentoxide at concentrations of 0, 2, 4, 8, 16, or 32 mg/m(3) by inhalation, 6 hours per day, 5 days per week for 16 days. Three males in the 32 mg/m(3) group died before the end of the study. Mean body weights of males and females exposed to 8 mg/m(3) or greater were less than those of the chamber controls. Clinical findings included rapid respiration and hypoactivity in rats exposed to 16 or 32 mg/m(3). Relative lung weights of 4 mg/m(3) or greater males and 2 mg/m(3) or greater females were significantly greater than those of the chamber controls. Lavage fluid analysis indicated an inflammatory response in the lung that was either directly mediated by vanadium pentoxide or was secondary to lung damage induced by vanadium pentoxide exposure. 16-DAY STUDY IN MICE: Groups of five male and five female mice were exposed to particulate aerosols of vanadium pentoxide at concentrations of 0, 2, 4, 8, 16, or 32 mg/m(3) by inhalation, 6 hours per day, 5 days per week for 16 days. All males exposed to 32 mg/m(3) and one 8 mg/m(3) male died or were killed moribund before the end of the study. Mean body weights of 16 mg/m(3) males and 8 mg/m(3) or greater females were significantly less than those of the chamber controls, and the 32 mg/m(3) females lost weight during the study. Absolute and relative lung weights of 4 mg/m(3) or greater males and all exposed groups of females and liver weights of 16 mg/m(3) males were significantly greater than those of the chamber controls. The mediastinal lymph nodes were enlarged in 4, 8, and 16 mg/m(3) males and females, and lymphoid hyperplasia was confirmed histologically. Lavage fluid analysis indicated an inflammatory response in the lung that was either directly mediated by vanadium pentoxide or was secondary to lung damage induced by vanadium pentoxide exposure. 3-MONTH STUDY IN RATS: Groups of 10 male and 10 female rats were exposed to particulate aerosols of vanadium pentoxide at concentrations of 0, 1, 2, 4, 8, or 16 mg/m(3) by inhalation, 6 hours per day, 5 days per week for 3 months. Seven males and three females exposed to 16 mg/m(3) died during the study. Mean body weights were significantly less in males exposed to 4 mg/m(3) or greater and in females exposed to 16 mg/m(3). Abnormal breathing, thinness, lethargy, abnormal posture, and ruffled fur were observed in rats exposed to 16 mg/m(3). Hematology results indicated that exposure of rats to vanadium pentoxide induced a microcytic erythrocytosis in males and females. Absolute and relative lung weights were significantly greater for 4 mg/m(3) or greater males and females than for the chamber controls as were the relative lung weights of 2 mg/m(3) males. The estrous cycle of females exposed to 8 mg/m(3) was significantly longer than that of the chamber control group, and the number of cycling females in the 16 mg/m(3) group was reduced. The incidences of several nonneoplastic lesions of the lung and nose were significantly increased in males and females exposed to 2 mg/m(3) or greater. Data from pulmonary function analyses indicated that a restrictive lung disease was present in male and female rats exposed to 4 mg/m(3) or greater, while an obstructive lung disease was present only in the 16 mg/m(3) groups. 3-MONTH STUDY IN MICE: Groups of 10 male and 10 female mice were exposed to particulate aerosols of vanadium pentoxide at concentrations of 0, 1, 2, 4, 8, or 16 mg/m(3) by inhalation, 6 hours per day, 5 days per week for 3 months. One male exposed to 16 mg/m(3) died before the end of the study. Mean body weights of 8 and 16 mg/m(3) males and 4 mg/m(3) or greater females were significantly less than those of the chamber controls. Absolute and relative lung weights of males and females exposed to 4 mg/m(3) or greater were significantly greater than those of the chamber controls. The epididymal spermatozoal motility of males exposed to 8 or 16 mg/m(3) was significantly decreased. Some mice exposed to 2 or 4 mg/m(3) had inflammation of the lung, and all mice exposed to 8 or 16 mg/m(3) had inflammation and epithelial hyperplasia of the lung. 16-DAY SPECIAL STUDY IN RATS: Groups of 60 female rats were exposed to particulate aerosols of vanadium pentoxide at concentrations of 0, 1, or 2 mg/m(3) and groups of 40 female rats were exposed to 4 mg/m(3) by inhalation, 6 hours per day, 5 days per week for 16 days. Alveolar and bronchiolar epithelial hyperplasia was observed in most rats exposed to 2 or 4 mg/m(3) on days 6 and 13. Histiocytic infiltration and inflammation occurred in a time- and concentration related manner. Cell turnover rates were increased in the terminal bronchioles on days 6 and 13 and in the alveoli in the 4 mg/m(3) group on day 6 and in all exposed groups on day 13. Assessment of lung vanadium concentrations suggested deposition and clearance exhibited linear kinetics over the exposure range studied. Lung clearance half-times ranged from 4.42 to 4.96 days. 16-DAY SPECIAL STUDY IN MICE: Groups of 60 female mice were exposed to particulate aerosols of vanadium pentoxide at concentrations of 0, 2, or 4 mg/m(3) and groups of 40 female mice were exposed to 8 mg/m(3) by inhalation, 6 hours per day, 5 days per week for 16 days. Alveolar and bronchiolar epithelial hyperplasia occurred with similar incidences and severities among the exposed groups on days 6 and 13, and time- and concentration-related increases in the incidences of interstitial inflammation and histiocytic infiltration also occurred in these groups. Cell turnover rates were increased in the terminal bronchioles on day 6 and remained greater than those of the chamber controls on day 13. In the alveoli, cell turnover rates were increased in an exposure concentration-related manner on day 13; cell turnover rates were increased only in the 8 mg/m(3) group on day 6. Assessment of lung vanadium concentrations suggested deposition and clearance exhibited linear kinetics over the exposure range studied. Lung clearance half times ranged from 2.40 to 2.55 days. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were exposed to particulate aerosols of vanadium pentoxide at concentrations of 0, 0.5, 1, or 2 mg/m(3) by inhalation, 6 hours per day, 5 days per week for 104 weeks. Survival and body weights of males and females were generally similar to those of the chamber controls. Mean body weights of females exposed to 2 mg/m(3) were less than those of the chamber controls throughout the study. Alveolar/bronchiolar neoplasms were present in exposed groups of male rats, and the incidences often exceeded the historical control ranges. Alveolar/bronchiolar adenomas were present in 0.5 and 1 mg/m(3) females; one 2 mg/m(3) female also had an alveolar/bronchiolar carcinoma. The incidence of alveolar/bronchiolar adenoma in the 0.5 mg/m(3) group was at the upper end of the historical control ranges. Nonneoplastic lesions related to vanadium pentoxide exposure occurred in the respiratory system (lung, larynx, and nose) of male and female rats, and the severities of these lesions generally increased with increasing exposure concentration. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice were exposed to particulate aerosols of vanadium pentoxide at concentrations of 0, 1, 2, or 4 mg/m(3) by inhalation, 6 hours per day, 5 days per week for 104 weeks. Survival of 4 mg/m(3) males was significantly less than that of the chamber controls. Mean body weights of 4 mg/m(3) males and all exposed groups of females were generally less than those of the chamber controls throughout the study, and those of males exposed to 2 mg/m(3) were less from week 85 to the end of the study. Many mice exposed to vanadium pentoxide were thin, and abnormal breathing was observed in some mice, particularly those exposed to 2 or 4 mg/m(3). The incidences of alveolar/bronchiolar neoplasms were significantly increased in all groups of exposed males and females. Nonneoplastic lesions related to vanadium pentoxide exposure occurred in the respiratory system (lung, larynx, and nose) of male and female mice, and the severities of these lesions generally increased with increasing exposure concentration. Bronchial lymph node hyperplasia was present in many exposed females. MOLECULAR ONCOLOGY STUDIES: K ras codon 12 mutation and loss of heterozygosity on chromosome 6 were detected in vanadium pentoxide-induced alveolar/bronchiolar carcinomas from mice. GENETIC TOXICOLOGY: Vanadium pentoxide was not mutagenic in Salmonella typhimurium strain TA97, TA98, TA100, TA102, or TA1535, with or without induced rat or hamster liver S9 enzymes. CONCLUSIONS: Under the conditions of this 2-year inhalation study, there was some evidence of carcinogenic activity of vanadium pentoxide in male F344/N rats and equivocal evidence of carcinogenic activity of vanadium pentoxide in female F344/Nrats based on the occurrence of alveolar/bronchiolar neoplasms. There was clear evidence of carcinogenic activity of vanadium pentoxide in male and female B6C3F1 mice based on increased incidences of alveolar/bronchiolar neoplasms. (ABSTRACT TRUNCATED) PMID- 12533745 TI - Toxicology and carcinogenesis study of chloral hydrate (ad libitum and dietary controlled) (CAS no. 302-17-0) in male B6C3F1 mice (gavage study). AB - [structure: see text] Chloral hydrate is used medically as a sedative or hypnotic and as a rubefacient in topical preparations, and it is often given to children as a sedative during dental and other medical procedures. Chloral hydrate is used as a central nervous system depressant and sedative in veterinary medicine and as a general anesthetic in cattle and horses. It is a byproduct of the chlorination of water and has been detected in plant effluent after the bleaching of softwood pulp. Chloral, the anhydrous form of chloral hydrate, is used as a synthetic intermediate in the production of insecticides and herbicides. Chloral hydrate was nominated for study by the Food and Drug Administration based upon widespread human exposure and its potential hepatotoxicity and the toxicity of related chemicals. A dietary control component was incorporated in response to concerns within the regulatory community relating to increased background neoplasm incidences in rodent strains used for toxicity testing and to the proposed use of dietary restriction to control background neoplasm incidence in rodent cancer studies. Male B6C3F1 mice (ad libitum-fed or dietary-controlled) received chloral hydrate (99% pure) by gavage for 2 years. 2-YEAR STUDY IN MALE MICE: Groups of 120 male mice received chloral hydrate in distilled water by gavage at doses of 0, 25, 50, or 100 mg/kg 5 days per week for 104 to 105 weeks. Each dose group was divided into two dietary groups of 60 mice. The ad libitum-fed mice had free access to feed, and the dietary-controlled mice received feed in measured daily amounts calculated to maintain body weight on a previously computed idealized body weight curve. Twelve mice from each diet and dose group were evaluated at 15 months. SURVIVAL, FEED CONSUMPTION, AND BODY WEIGHTS: Survival of dosed groups of ad libitum-fed and dietary-controlled mice was similar to that of the corresponding vehicle controls. When compared to the ad libitum-fed groups, dietary control significantly increased survival in the vehicle controls and 25 and 50 mg/kg groups. Mean body weights of all dosed groups were similar to those of the vehicle control groups throughout the study. The dietary-controlled mice were successfully maintained at or near their target idealized body weights. There was less individual variation in body weights in the dietary-controlled groups than in the corresponding ad libitum-fed groups. Feed consumption by 25 and 50 mg/kg ad libitum-fed mice was generally similar to that by the vehicle controls throughout the study. Feed consumption by 100 mg/kg ad libitum-fed mice was slightly less than that by the vehicle controls throughout the study. HEPATIC ENZYME ANALYSIS: Chloral hydrate did not significantly induce either lauric acid 4-hydroxylase activity or CYP4A immunoreactive protein in any of the dosed groups of ad libitum-fed mice. However, 100 mg/kg did significantly induce both lauric acid 4-hydroxylase activity and CYP4A immunoreactive protein in the dietary controlled mice. Moreover, the induction response profile of CYP4A was similar to the increase in the incidence of liver neoplasms at 2 years in the dietary controlled mice with the major effect occurring in the 100 mg/kg group. The serum enzymes alanine aminotransferase, amylase, aspartate aminotransferase, and lactate dehydrogenase were also assayed at 2 years. In the ad libitum-fed groups there was a significant increase in aspartate aminotransferase activity in the 50 mg/kg group. There were no other significant effects in any dosed group, but in general the dietary-controlled groups exhibited lower values than the corresponding ad libitum-fed groups. ORGAN WEIGHTS AND PATHOLOGY FINDINGS: The heart weight of ad libitum-fed male mice administered 100 mg/kg and the kidney weights of 50 and 100 mg/kg ad libitum-fed mice were significantly less than those of the vehicle controls at 2 years. The liver weights of all dosed groups of ad libitum-fed and dietary-controlled mice were greater than those of the vehicle control groups at 2 years, but the increases were not statistically significant. The incidence of hepatocellular adenoma or carcinoma (combined) in ad libitum-fed mice administered 25 mg/kg was significantly greater than that in the vehicle controls at 2 years. The incidences of hepatocellular carcinoma and of hepatocellular adenoma or carcinoma (combined) occurred with positive trends in dietary-controlled male mice at 2 years, and the incidence of hepatocellular carcinoma in 100 mg/kg dietary-controlled mice was significantly increased. CONCLUSIONS: Under the conditions used in this 2-year gavage study, there was some evidence of carcinogenic activity of chloral hydrate in male B6C3F1 mice based on increased incidences of hepatocellular adenoma or carcinoma (combined) in ad libitum-fed mice and on increased incidences of hepatocellular carcinoma in dietary-controlled mice. In the dietary-controlled mice, induction of enzymes associated with peroxisome proliferation was observed at higher doses. PMID- 12533746 TI - P2 receptors: theoretical background for the use in clinical practice. AB - Here we review the results of physiological, pharmacological, and pathophysiological studies of ATP receptors, P2 type receptors. Characteristics of two families of P2 receptors (P2X and P2Y) are presented and the possibility of using preparations affecting these receptors is discussed. PMID- 12533747 TI - Independence of changes in right atrial pressure and central venous pressure. AB - Dynamics of changes in right-atrial pressure and venous pressure measured in the inferior and superior vena cava at their orifices (central venous pressure) after bolus injection of 20 ml physiological saline or epinephrine (5.0 mg/kg) was studied in acute experiments on cats. The initial pressure in the right atrium was equal to that in caval veins. Pressor stimuli either increased or decreased the right atrial pressure, but always increased blood pressure in the caval veins. Moreover, right atrial pressure returned to the initial level more rapidly compared to that in caval veins. Our results suggest that the dynamics of the right-atrial pressure does not reflect the shifts in the central venous pressure. PMID- 12533749 TI - Reactions of hemopoietic granulocytic stem in hypoxia of different severity. AB - We studied the reactions of granulocytic hemopoietic stem after acute hypoxia and during the development of posthypoxic encephalopathy. Damage to brain structures was associated with intensification of the bone marrow hemopoiesis due to activation of hemopoiesis-inducing microenvironment and more intense formation of hemopoietic islets, despite reduced proliferative capacity of granulocytic precursors. PMID- 12533748 TI - Adaptation to heat limits stress-induced activation of caspases in the thymus. AB - We demonstrated selective activation of caspases in the thymus during heat shock, i.e. primary activation of initiator caspase 9 (but not caspase 8) and effector caspase 3 (but not caspase 6). Preadaptation to heat improved animal survival after heat shock and reduced heat shock-induced activation of both initiator and effector caspases. Hence, adaptation to heat produced an antiapoptotic effect, which was not selective towards receptor-dependent or mitochondrial pathway of the caspase cascade activation. PMID- 12533750 TI - Adaptation to stress improves resistance to gastric damage during acute stress in Wistar rats and decreases resistance in August rats: role of serotonin. AB - In August rats more resistant to acute stress-induced gastric damage than Wistar rats, preadaptation to nondamaging stress exposure did not prevent damage and even potentiated these damages. By contrast, in Wistar rats such adaptation decreased gastric damage caused by acute stress. Higher initial resistance of August rats to stress damage was associated with higher serotonin level and lower norepinephrine/serotonin ratio in the gastric mucosa than in Wistar rats. The negative effect of adaptation in August rats was associated with decreased serotonin level and increased norepinephrine/serotonin ratio in the stomach during stress. In Wistar rats exposed to stress the protective effect of adaptation was associated with an increase of serotonin content and a decrease of the norepinephrine/serotonin ratio in the stomach. Hence, the degree of resistance to stress-induced gastric damage can be due to genetically determined serotonin level and norepinephrine/serotonin ratio in the stomach. PMID- 12533751 TI - Free-radical lipid oxidation in intergeneric and interspecific sturgeon hybrids at early stages of ontogeny. AB - The intensity of free-radical lipid oxidation was evaluated in total homogenates of intergeneric (great sturgeon with barbel sturgeon) and interspecific (Russian sturgeon with barbel sturgeon) hybrid fry and their parental species. Activity of the antioxidant defense system in intergeneric fry was higher than in interspecific fry, which should be taken into consideration during selection of objects for intensive aquaculture. PMID- 12533752 TI - Biochemical characteristics and water exchange in the surface layer of human joint cartilage. AB - We compared the contents of collagen, glycosaminoglycans, and various forms of water in the surface layer and whole tissue of joint cartilages of different localization. It was found that the surface layer is characterized by reduced content of glycosaminoglycans compared to the whole tissue and higher water holding capacity due high content of bound water. PMID- 12533753 TI - NO-inhibiting and vasotropic activity of some compounds with thioamidine group. AB - Using the method of electron paramagnetic spectroscopy we demonstrated that thiazine-thiazoline compounds and aminoethyl isothiourea containing the thioamidine group inhibit NO production in the liver of endotoxin-treated mice. Injection of these agents to anesthetized rats increased arterial pressure and enhanced respiration rate. This effect probably reflects inhibition of not only inducible, but also the constitutive synthesis of NO by compounds with thioamidine group. PMID- 12533754 TI - Hepatoprotective activity of 1-ethoxysilatrane and 1-isopropoxygermatrane. AB - 1-Ethoxysilatrane and 1-isopropoxygermatrane stimulated liver regeneration via activation of some components of the protein-synthesizing complex, in particular, aminoacyl-tRNA synthetases. Acylating activity of aminoacyl-tRNA synthetases increased. Possible mechanisms of these changes are discussed. PMID- 12533755 TI - Effect of lipid matrix and cytoskeleton proteins on Ca2+-activated K+ channels in erythrocytes of alcoholic and II type diabetes mellitus patients. AB - We studied the effect of changes in erythrocyte volume and irreversible thermal denaturation of cytoskeleton proteins and lipid matrix on activity of Ca(2+) activated K+ channels in erythrocytes of alcoholic and patients with II type diabetes mellitus. Changes in Ca(2+)-dependent potassium permeability of erythrocyte membrane in alcoholic patients and patients with II type diabetes mellitus are related to modification of cytoskeleton, rather than to changes in lipid matrix. PMID- 12533756 TI - Effect of mifegin on the content of cyclic nucleotides in the cervical myometrium in full-term pregnancy. AB - Cervical myometrium was examined in 22 pregnant women at 39-41 weeks gestation at risk of labor abnormalities. The patients received mifegin for preparing to labor. A new component in the mechanism of mifegin effect on the uterine cervix in full-term pregnancy was revealed: mifegin elevated the content of cAMP by 50% and almost 2-fold increased the cAMP/cGMP ratio compared to the initial level. PMID- 12533757 TI - Relationship between alcohol motivation and ethanol sensitivity and blood and brain catecholamine content in the progeny of rats preferring and nonpreferring alcohol. AB - A system of neurochemical signs indicating predisposition to high ethanol consumption is formed in the third generation descendants of ethanol-preferring male rats. Risk-group males are detected (with 95% probability) by the content of dopamine and norepinephrine and their ratio in the brain. Alcohol motivation depends on blood epinephrine content reflecting individual sensitivity to stress. PMID- 12533758 TI - Protective effect of enterosgel on rat liver lysosomes during cytostatic treatment. AB - Polychemotherapy with a complex of cytostatics (cyclophosphamide, doxorubicin, vincristine, prednisolone) induces progressive damage to hepatocyte membranes, which manifested in labilization of lysosomes and activation of lysosomal enzymes and serum transaminases. Enterosgel stabilized liver lysosomes and reduced manifestation of hepatocyte cytolysis. PMID- 12533759 TI - Effect of in vivo and in vitro stimulation of delta1-opioid receptors on myocardial resistance to arrhythmogenic action of ischemia and reperfusion. AB - Preliminary intravenous injection of peptide agonist of delta1-opioid receptors DPDPE (0.5 mg/kg) decreased the incidence of occlusion (10 min) and reperfusion (10 min) arrhythmias in rats. By contrast, delta2-opioid receptor agonist DSLET produced no effect on the incidence of arrhythmias provoked by coronary occlusion and reperfusion. Preliminary injection of selective delta-receptor antagonist ICI 174,864 (2.5 mg/kg) or TIPP[y] (0.5 mg/kg) completely abolished the antiarrhythmic effect of DPDPE. Stimulation of cardiac delta1-opioid receptors with DPDPE added to perfusion saline in concentrations of 0.1 and 0.5 mg/liter decreased the incidence of reperfusion arrhythmias. Addition of DPDPE to perfusion saline in a concentration of 0.1 mg/liter prevented reoxygenation destruction of cardiomyocytes. By contrast, no cardioprotective effect of this peptide was observed at a concentration of 0.5 mg/liter in perfusion saline or when it was injected intravenously. It is concluded that the cardioprotective and antiarrhythmic effects of DPDPE are caused by activation of cardiac delta1-opioid receptors. PMID- 12533760 TI - Conjugates of aberrant gangliosides in antiglioma vaccine: toxicological assay. AB - We studied sterility and toxicity of vaccine LS1 containing aberrant gangliosides isolated from brain bioptates of 48 patients with gliomas of different malignancy and covalently bound to keyhole limpet hemocyanin. The vaccine was safe. This preparation produced no side effects in experimental animals. Our findings substantiated the necessity of father development of this method of vaccination. The vaccine should undergo clinical tests in patients with malignant gliomas. PMID- 12533761 TI - Photoprotective activity of melanin preparations in human skin exposed to UV irradiation: dependence on previous photoexposure. AB - Photoprotective effects of three melanin preparations (from black yeast fungi and Sepia sp.) were studied. These preparations in aqueous solutions (5 g/ml, dark exposure for 7 days) demonstrated high photomodification capacity upon exposure to visible light in doses of up to 1.8 kJ/m2. Preliminary exposure of these solutions to visible light in a dose of 360 kJ/m2 notably decreased the photoprotective effect of melanins during UV exposure of the skin treated with these solutions (at UV dose of 3.4 kJ/m2). This necessitates empirical selection of the dose and storage condition of melanin preparations for attaining the optimal photoprotective effect. PMID- 12533762 TI - Comparative study of Saccharomyces cerevisiae LPS. AB - LPS of yeast strains producing human epidermal growth factor were studied. Experiments demonstrated the absence of essential differences in the characteristics of these LPS and LPS of nonrecombinant Saccharomyces cerevisiae strains, which allowed us to develop a universal complex technology of simultaneous preparation of heterologous proteins and highly active immunomodulating LPS. PMID- 12533763 TI - Astrakhan scrub typhus: time course of infectious process according to electron microscopy findings. AB - Structural changes in the liver and spleen of albino mice with Astrakhan scrub typhus were studied by electron microscopy. Rickettsia invasion and formation of granulomas induced structural (destructive) and metabolic changes in hepatocytes. Rickettsia were degraded in cytophagosomes and cytolysophagosomes of hepatic macrophages (Kupffer cells) and blood capillary endotheliocytes. In the spleen rickettsia were seen in the extracellular spaces and in various cell populations. PMID- 12533764 TI - Immune reactions in different mouse strains. AB - The responses of immunocompetent cells to thymus-dependent antigen differ in mice of different strains. Immunization stimulated phagocytic activity of peritoneal macrophages in CBA/CaLac, DBA/2, and BALB/c mice and suppressed it in CC57W mice. By the formation of antibody-producing cells in the spleen in response to thymus dependent antigen DBA/2 and CBA/CaLac mice can be classified as high responders, BALB/c mice as medium-responders, and C57Bl/6 and CC57W mice as low responders. PMID- 12533765 TI - Use of nested PCR for differential diagnosis of falciparum malaria reinfection and relapse in drug-resistant patients. AB - Microscopic examination does not allow differentiation of drug-resistant P. falciparum infection relapse from reinfection. However, this differential diagnosis is essential for adequate therapy. Three highly polymorphic P. falciparum genes (msp1, msp2, and glurp) and their alleles reflecting the structural state of these genes were used as genetic markers for differential diagnosis by PCR with internal primers. In 27 patients the characteristics of these alleles were identical before treatment with artersunate and during repeated manifestation of symptoms 14-28 days after the end of therapy, which attested to malaria relapses. In 24 patients the structure of these allele before mefloquine therapy and during repeated manifestation of the symptoms after 2-3 months was different, which attested to reinfection. PMID- 12533766 TI - Phenotypic characteristics of lewis lung carcinoma cells. AB - We studied adhesive properties and physiological activity in vivo of cells from Lewis lung carcinoma and its metastases. These cells differed in tumorogenic activity and metastatic potential in the syngeneic system. In vivo non metastasizing cells are characterized by a lower content of surface lectins to tetrasaccharides SiaLex [Neu5Aca2-3Galb1-4(Fuca1-3) GlcNAcb] and SiaLea [Neu5Aca2 3Galb1-3(Fuca1-4)GlcNAcb] and trisaccharide HSO3Lex [HSO32-3Galb1-4(Fuca1 3)GlcNAcb] compared to cells forming metastases in the syngeneic system. Metastatic cells with low tumorogenic activity weakly expressed lectins to disaccharide ligands 6-SiaLac [Neu5Aca2-6Galb1-4Glc], 6-HSO3LacNAc, and A-di [GalNAca 1-3Galb] and trisaccharides H-type 1 [Fuca1-2Galb1-3GlcNAc and Lex [Fuca1-3(Galb 1-4)GlcNAc] compared to cells that initiated tumor formation in the syngeneic system (similarly to transplanted tumors). We hypothesized that cell receptors to these carbohydrate determinates are involved in the development and growth of primary tumors, while lectins to SiaLex, SiaLea, and HSO3Lex play a role in the progress of tumor process and metastasizing. PMID- 12533767 TI - 6alpha-Methyl-16alpha,17alpha-cyclohexane progesterone and progesterone inhibit growth of doxorubicin-sensitive MCF-7 and HeLa tumor cells. AB - Temporal and concentration dependencies of the effects of gestagens (6alpha methylpentarane and progesterone) and cytostatic doxorubicin on proliferation of MCF-7 and HeLa tumor cells was studied using(3)H-thymidine test. Gestagens produced the maximum inhibitory effect of on cell proliferation in a concentration of 10(-5)M; the effect developed on day 6 of incubation. 6 alpha Methylpentarane in a concentration of 10(-8)inhibited proliferation of HeLa cells more effectively than progesterone (p<0.05). In experiments with combined treatment of doxorubicin-sensitive MCF-7 and HeLa cells, progesterone in a concentration of 10(-7)M attenuated the cytostatic effect of doxorubicin (p<0.05), while 6alpha-methylpentarane in the studied concentrations did not modulate it. PMID- 12533768 TI - Effects of Livagen peptide on chromatin activation in lymphocytes from old people. AB - We studied the effects of the synthetic peptide Livagen on activity of ribosomal genes, denaturation parameters of heterochromatin, polymorphism of structural C heterochromatin, and variability of facultative heterochromatin in lymphocytes from old people. Livagen induced activation of ribosomal genes, decondensation of pericentromeric structural heterochromatin, and release of genes repressed due to age-related condensation of euchromatic regions in chromosomes. Our results indicate that Livagen causes de-heterochromatinization (activation) of chromatin, which is realized via modification of heterochromatin and heterochromatinized regions in chromosomes from old people. PMID- 12533769 TI - In vitro response of mitochondrial succinate oxidase system to epinephrine in human blood lymphocytes from health individuals and patients with neurocirculatory dystonia. AB - Activities of succinate dehydrogenase (SDH) and alpha-glycerophosphate dehydrogenase (hyaloplasmic and mitochondrial: alpha-GPDHH and alpha-GPDHM) in peripheral blood lymphocytes, and the response of SDH activity to exogenous epinephrine in vitro (the epinephrine test) were studied in 20 healthy subjects and 46 patients with hypertensive neurocirculatory dystonia. Activities of SDH, alpha-GPDHH, and alpha-GPDHM in blood lymphocytes and SDH adrenoreactivity in epinephrine test were higher in patients than in healthy controls. Treatment with hypotensive agents (isradipin and pyrroxan) moderated adrenoreactivity. Phytotherapy normalized the baseline activities of succinate oxidase system and alpha-glycerophosphate pathway in blood lymphocytes. PMID- 12533770 TI - Trophoblastic beta1-glycoprotein and hemostasis system in pregnant women with antiphospholipid syndrome. AB - Clinical and laboratory studies were carried out in 38 pregnant women with antiphospholipid syndrome. Increased functional activity of platelets and decreased protein-producing function of the placenta were observed starting from the early terms of gestation. These disorders were followed by the development of hypercoagulation in the plasma component of hemostasis, appearance of intravascular blood clotting markers, and inhibition of AT III and protein C. This led to the progress of disorders in the microcirculatory bed, fetoplacental insufficiency, decrease in trophoblastic beta1-glycoprotein level, chronic hypoxia, and fetal death. Infection accelerated this process. Measurements of trophoblastic beta1-glycoprotein every 2 weeks help to diagnose fetoplacental disorders, predict the course of pregnancy, and evaluate the efficiency of drug therapy. PMID- 12533771 TI - Effects of acute and chronic caffeine intake on intravenous self-administration of morphine in two rat strains. AB - Effects of acute and chronic caffeine intake on the level and pattern of morphine self-administration behavior in WAG/G and Fisher-344 rats were studied. Both acute and chronic caffeine intake decreased morphine self-administration only in WAG/G rats, which attested to increased sensitivity of these rats to reinforcing effects of morphine. Possible relationship between the observed changes and increased anxiety in rats receiving caffeine is discussed. PMID- 12533772 TI - Measurements of time and space as manifestation of spatiotemporal organization of reflection. AB - We studied measurements of the individual minute and individual decimeter. Individual time and length measures were reproduced separately or in combination (drawing 1 dm over 1 min). The measurements of time and space as coordinates of a vector for the space-and-time image are a single system of spatiotemporal organization of reflection. We revealed individual differences in the measurement of time and space. The individual minute was more variable than the individual decimeter. PMID- 12533773 TI - Expression of neuronal and inducible nitric oxide synthases in the thymus under normal conditions and after administration of bacterial endotoxin. AB - The topography of thymocytes expressing neuronal and inducible nitric oxide synthases and changes in the content of luminescent immunoreactive products in these cells after intraperitoneal injection of bacterial lipopolysaccharide were studied by double immunohistochemical labeling. Under normal conditions neuronal nitric oxide synthase-immunopositive cells formed a wide network in thymus medulla (except for perivascular regions). Inducible nitric oxide synthase was expressed in single cells at the corticomedullary boundary. Lipopolysaccharide markedly increased the intensity of luminescence and number of inducible nitric oxide synthase-immunoreactive cells. However, this agent sharply decreased the intensity of luminescence in neuronal nitric oxide synthase-immunopositive cells of the stroma. Our results indicate that neuronal and inducible nitric oxide synthases are synthesized in various stromal cells of the thymus. Expression of these enzyme isoforms undergoes opposite changes during inflammation. PMID- 12533774 TI - Ultrastructural and morphometric characteristics of nerve cells and myelinated fibers in the cerebral cortex after chronic exposure to natural gas containing hydrogen sulfide in low concentrations. AB - We studied ultrastructural and morphometric characteristics of nerve cells and myelinated fibers in the cerebral cortex after chronic exposure to natural gas containing hydrogen sulfide in low concentrations. Radioisotope assay revealed activation of protein synthesis in nerve cells after chronic exposure to natural hydrogen sulfide-containing gas in low concentrations (10 mg/m(3)by H2S) for 2 weeks. After 1 month the ultrastructure of myelinated fibers was characterized by sectorial loosening and demyelination. PMID- 12533775 TI - Pathomorphological changes in hyaline cartilage during focal persistent infection. AB - We studied morphological changes in hyaline cartilages of different localization in adult male rabbits with focal persistent infection produced by Staphylococcus aureus strain 209. Local chronic inflammation produced systemic atrophic and degenerative changes in the cartilage tissue. The severity of pathological changes depended on genetically determined repair and metabolic activity of the cartilage tissue. PMID- 12533776 TI - Liposome-entrapped polymerases as models for microscale/nanoscale bioreactors. PMID- 12533777 TI - Real-time observation of lipoplex formation and interaction with anionic bilayer vesicles. AB - A novel development has allowed for the direct observation of single, pairwise interactions of linear DNA with cationic vesicles and of DNA-cationic lipid complexes with anionic vesicles. A new cationic phospholipid derivative, l,2 dioleoyl-sn-glycero-3-ethylphosphocholine, was used to prepare giant bilayer vesicles and to form DNA-cationic lipid complexes (lipoplexes). The cationic vesicles were electrophoretically maneuvered into contact with DNA, and similarly, complexes were brought into contact with anionic phospholipid vesicles composed of dioleoylphosphatidylglycerol (DOPG; 100%), DOPG/dioleoylphosphatidylethanolamine (DOPE; 1:1) or DOPG/dioleoylphosphatidylcholine (DOPC; 1:1). Video fluorescence microscopy revealed that upon contact with phospholipid anionic vesicles, lipoplexes exhibited four different types of behavior: adhesion, vesicle rupture, membrane perforation (manifested as vesicle shrinkage and/or content loss), and expansion of DNA (which was always concomitant with membrane perforation.) In one instance, the lipoplex was injected into the target vesicle just prior to DNA expansion. In all other instances, the DNA expanded over the outer surface of the vesicle, and expansion was faster, the larger the area of vesicle over which it expanded. Given the likelihood of incorporation of cellular anionic lipids into lipoplexes, the expansion of the DNA could be important in DNA release during cell transfection. Upon contact with naked DNA, giant cationic vesicles usually ruptured and condensed the DNA into a small particle. Contact of cationic vesicles that were partially coated with DNA usually caused the DNA to wrap around the vesicle, leading to vesicle rupture, vesicle fusion (with other attached vesicles or lipid aggregates), or simply cessation of movement. These behaviors clearly indicated that both DNA and vesicles could be partly or fully covered by the other, thus modifying surface charges, which, among others, allowed adhesion of DNA-coated vesicles with uncoated vesicles and of lipid coated DNA with uncoated DNA. PMID- 12533778 TI - Cardiolipin regulates the activity of the reconstituted mitochondrial calcium uniporter by modifying the structure of the liposome bilayer. AB - Reconstitution of mitochondrial calcium transport activity requires the incorporation of membrane proteins into a lipidic ambient. Calcium uptake has been measured previously using Cytochrome oxidase vesicles. The enrichment of these vesicles with cardiolipin, an acidic phospholipid that is found only in the inner mitochondrial membrane of eukaryotic cells, strongly inhibits calcium transport, in remarkable contrast with the activation effect that cardiolipin exerts upon other mitochondrial transporters and enzymes. The relation of the inactivation of calcium transport to the physical state of the bilayer was studied by following the polarization changes of 1,6-diphenyl-1,3,5-hexatriene (DPH) and by flow cytometry in the cardiolipin-enriched liposomes with incorporated mitochondrial solubilized proteins. Non-bilayer molecular arrangements in the cardiolipin-supplemented liposomes, detected by flow cytometry, may produce the fluidity changes observed by fluorescence polarization of DPH. Fluidity changes correlate with the abolition of calcium uptake, but have no effect on the establishment of a membrane potential in the vesicles required for calcium transport activity. Changes in the membrane structure and uniporter function are observed in the combined presence of cardiolipin and calcium leading to a modified lipid configuration. PMID- 12533779 TI - The resting potential of mouse Leydig cells: role of an electrogenic Na+/K+ pump. AB - Resting potentials (Vm) were measured in mouse Leydig cells, using the whole-cell patch-clamp technique. In contrast to conventional microelectrode measurements, where a biphasic potential was observed, we recorded a stable Vm around -32.2 +/- 1.2 mV (mean +/- SEM, n = 159), at 25 degrees C, and an input resistance larger than 2.7 x 109 W. Although Vm is sensitive to changes in the extracellular concentrations of potassium and chloride, the relationship between Vm and these ions' concentrations cannot be described by either the Goldman-Hodgkin-Katz or the Nernst equation. Perifusing cells with potassium-free solution or 10?3 M ouabain induced a marked depolarization averaging 20.1 +/- 3.2 mV (n = 9) and 23.1 +/- 2.8 mV, (n = 7), respectively. Removal of potassium or addition of ouabain with the cell voltage-clamped at its Vm, resulted in an inwardly directed current, due to inhibition of the Na+K+ATPase. The pump current increased with temperature with a Q10 coefficient of 2.3 and had an average value of -6.5 +/- 0.4 pA (n = 21) at 25 degrees C. Vm also varied strongly with temperature, reaching values as low as -9.2 +/- 1.2 mV (n = 22) at 15 degrees C. Taking the pump current at 25 degrees C and a minimum estimate for the membrane input resistance, we can see that the Na+K+ATPase could directly contribute with 17.7 mV to the Vm of Leydig cells, which is a major fraction of the ?32.2 +/- 1.2 mV (n = 159) observed. PMID- 12533780 TI - Basolateral Cl- transport is stimulated by terbutaline in adult rat alveolar epithelial cells. AB - Stimulation of adult rat alveolar epithelial cells with terbutaline was previously shown to activate Cl- channels in the apical membrane. In this study, we show that terbutaline stimulates net transepithelial (apical-to-basolateral) Cl- absorption from 0.19 +/- 0.13 to 1.43 +/- 0.31 mmol x cm-2 x hr-1. Terbutaline also increases net Cl- efflux across the basolateral membrane under conditions where an outward [K+] gradient exists and the membrane voltage is clamped at zero mV. When the [K+] gradient is eliminated, the effect of terbutaline on net Cl- efflux is inhibited to the extent that no significant Cl- efflux can be detected across the basolateral membrane. RT-PCR experiments detected mRNA for three KCl cotransport isoforms (KCC1, KCC3 and KCC4) in monolayer cultures of alveolar epithelial cells. Western blot analysis using antibodies to the four cloned isoforms of KCl cotransporters revealed the presence of KCC1 and KCC4 isoforms in monolayer cultures of these cells. These results provide evidence suggesting a role for KCl cotransport in terbutaline stimulated transepithelial Cl- absorption. PMID- 12533781 TI - The role of phosphatidylinositol 3-kinase (PI3K) in CO2 stimulation of the Na+/HCO3- cotransporter (NBC). AB - The basolateral Na+/HCO3- cotransporter (NBC) is the major pathway for bicarbonate reabsorption in the renal proximal tubule cells. The cotransporter activity is enhanced by 10% CO2. Phosphatidylinositol 3-kinase (PI3K) has been shown to regulate the function and trafficking of cellular proteins by promoting their translocation to the plasma membrane. Therefore, we sought to examine the role of PI3K in CO2-mediated stimulation of NBC activity in OK cells. Our studies showed that wortmannin, a well-characterized PI3K inhibitor, had no effect on baseline NBC activity but prevented the stimulatory effect of 10% CO2. This effect was concentration-dependent and time-dependent. Another inhibitor of PI3K, LY294002, also prevented the CO2-mediated increase in NBC activity. CO2 stimulation of the cotransporter was paralleled by an increase in PI3K enzyme activity and this effect was blocked by wortmannin. Biotinylation studies also showed that 10% CO2 increased the immunoreactive NBC in the basolateral membranes and this was prevented by wortmannin. We previously showed that 10% CO2 stimulation of NBC activity involves the Src family kinase pathway. In the current studies, CO2 stimulation significantly increased Src phosphorylation and this effect was abrogated by wortmannin. In summary, CO2 stimulation of NBC is mediated at least in part by increased immunoreactive NBC protein in the basolateral membrane, a process which requires the interaction of PI3K with Src family kinase. PMID- 12533782 TI - Signals that regulate GLUT4 translocation. PMID- 12533783 TI - Polarized expression of Shaker channels in epithelial cells. AB - The polarized distribution of ion channels into an apical or a basolateral domain is a fundamental feature of the transporting-epithelial phenotype. To study the molecular motifs of the channel that may serve as addressing signal(s), as well as the cellular mechanisms that interpret it and deliver the protein accordingly, we study the fate of transfected ShIR K+ channels (a non-inactivating Shaker channel) tagged with an HA epitope, as well as several other deletants and mutants. Surface expression is triggered by Ca2+-activated cell-cell contacts, through a cascade including a phospholipase C, a protein kinase C, and the cytoskeleton of actin and tubulin, and is partially impaired by suppressing N glycosylation with tunicamycin. Using domain-specific biotinylation we show that the channel is delivered preferentially to the basolateral domain thanks to a segment between amino acids 571 and 613, and is retained on the membrane surface due to a region involving the last three amino acids (threonine, aspartic acid, valine, TDV) of the COOH terminal. Its association with the cytoskeleton seems to take the form of a scaffold comprising actin, a-actinin, b-tubulin, mLin7 and CASK. We also observe that membrane expression of ShIR channels depends entirely on its sequence of amino acids and the conformation that the molecule may adopt, but not on its ability to translocate K+ across the membrane. PMID- 12533784 TI - Forskolin-induced clearance of the fluorescent dye sulforhodamine from rat parotid intralobular duct lumen: visualization of the secretory function under a confocal laser scanning microscope. AB - Cyclic AMP evokes fluid secretion with bicarbonate in exocrine ducts. Clearance of fluorescent dyes from rat parotid intralobular ducts by forskolin was visualized as a fluorescence change in the duct luminal space by optical sectioning under a confocal laser scanning microscope to clarify the secretory function in the ducts. When the isolated rat parotid intralobular duct segments were superfused with membrane-impermeable fluorescent dyes during the experimental period, fluorescent dyes were passively moved into the duct space. Forskolin and isobutylmethylxanthine decreased the fluorescence of anionic dye, sulforhodamine B, and neutral dye, dextran tetramethyl-rhodamine, in the duct space, suggesting that the forskolin-induced clearance of fluorescent dyes might be the result of fluid secretion in the ducts. Methazolamide inhibited a forskolin-induced sustained decrease in duct fluorescence and intracellular acidification. Low concentrations of external Cl?, DIDS, bumetanide and amiloride did not markedly inhibit a forskolin-induced decrease in duct fluorescence. These findings suggest that a major portion of the steady decrease in duct fluorescence by forskolin was related to intracellular HCO3? production, not the uptake mechanism of external Cl?. Glibenclamide, NPPB, DPC and DMA inhibited the forskolin-induced decrease. Forskolin evokes the clearance of fluorescent dyes from duct space possibly due to fluid secretion in rat parotid ducts, associated with secretion through CFTR and DPC-sensitive anion channels of carbonic anhydrase-dependent bicarbonate linked with the Na+/H+ exchange mechanism. PMID- 12533785 TI - Cl secretagogues reduce basolateral K permeability in the rabbit corneal epithelium. AB - The stromal-to-tear transport of Cl by the rabbit corneal epithelium is increased by pharmacological effectors (secretagogues) that raise cAMP. It is well established that such secretagogues increase the apical membrane permeability to Cl and thus facilitate the efflux of the anion. However, we and others have found that cAMP-elevating agents frequently decrease the transepithelial potential difference across the rabbit cornea. The mechanism underlying this latter phenomenon had not been characterized. In this report, transepithelial and microelectrode studies were combined with measurements of unidirectional fluxes of 36Cl, 22Na and 86Rb to show that secretagogues known to act via cAMP also decrease the K permeability of the basolateral membrane, which by cellular depolarization would decrease apical Cl secretion. This effect was increasingly pronounced as a function of concentration when agents (e.g., epinephrine, isoproterenol) were applied to the apical side of the preparations. The addition of these agonists to the basolateral bathing solution, or of forskolin to the apical side, solely elicited inhibitions of basolateral K permeability. It seems that apical Cl and basolateral K conductances are independently and inversely regulated by cAMP. The opposite effects that cAMP could have on fluid secretion and epithelial thickness, by increasing apical Cl permeability but decreasing basolateral K permeability, may serve as a mechanism to maintain epithelial thickness within a narrow range. PMID- 12533786 TI - Studying parents and grandparents to assess genetic contributions to early-onset disease. AB - Suppose DNA is available from affected individuals, their parents, and their grandparents. Particularly for early-onset diseases, maternally mediated genetic effects can play a role, because the mother determines the prenatal environment. The proposed maximum-likelihood approach for the detection of apparent transmission distortion treats the triad consisting of the affected individual and his or her two parents as the outcome, conditioning on grandparental mating types. Under a null model in which the allele under study does not confer susceptibility, either through linkage or directly, and when there are no maternally mediated genetic effects, conditional probabilities for specific triads are easily derived. A log-linear model permits a likelihood-ratio test (LRT) and allows the estimation of relative penetrances. The proposed approach is robust against genetic population stratification. Missing-data methods permit the inclusion of incomplete families, even if the missing person is the affected grandchild, as is the case when an induced abortion has followed the detection of a malformation. When screening multiple markers, one can begin by genotyping only the grandparents and the affected grandchildren. LRTs based on conditioning on grandparental mating types (i.e., ignoring the parents) have asymptotic relative efficiencies that are typically >150% (per family), compared with tests based on parents. A test for asymmetry in the number of copies carried by maternal versus paternal grandparents yields an LRT specific to maternal effects. One can then genotype the parents for only the genes that passed the initial screen. Conditioning on both the grandparents' and the affected grandchild's genotypes, a third log-linear model captures the remaining information, in an independent LRT for maternal effects. PMID- 12533787 TI - Estimation of haplotype frequencies, linkage-disequilibrium measures, and combination of haplotype copies in each pool by use of pooled DNA data. AB - Inference of haplotypes is important for many genetic approaches, including the process of assigning a phenotype to a genetic region. Usually, the population frequencies of haplotypes, as well as the diplotype configuration of each subject, are estimated from a set of genotypes of the subjects in a sample from the population. We have developed an algorithm to infer haplotype frequencies and the combination of haplotype copies in each pool by using pooled DNA data. The input data are the genotypes in pooled DNA samples, each of which contains the quantitative genotype data from one to six subjects. The algorithm infers by the maximum-likelihood method both frequencies of the haplotypes in the population and the combination of haplotype copies in each pool by an expectation maximization algorithm. The algorithm was implemented in the computer program LDPooled. We also used the bootstrap method to calculate the standard errors of the estimated haplotype frequencies. Using this program, we analyzed the published genotype data for the SAA (n=156), MTHFR (n=80), and NAT2 (n=116) genes, as well as the smoothelin gene (n=102). Our study has shown that the frequencies of major (frequency >0.1 in a population) haplotypes can be inferred rather accurately from the pooled DNA data by the maximum-likelihood method, although with some limitations. The estimated D and D' values had large variations except when the /D/ values were >0.1. The estimated linkage disequilibrium measure rho2 for 36 linked loci of the smoothelin gene when one- and two-subject pool protocols were used suggested that the gross pattern of the distribution of the measure can be reproduced using the two-subject pool data. PMID- 12533788 TI - Mutations in CHEK2 associated with prostate cancer risk. AB - The DNA-damage-signaling pathway has been implicated in all human cancers. However, the genetic defects and the mechanisms of this pathway in prostate carcinogenesis remain poorly understood. In this study, we analyzed CHEK2, the upstream regulator of p53 in the DNA-damage-signaling pathway, in several groups of patients with prostate cancer. A total of 28 (4.8%) germline CHEK2 mutations (16 of which were unique) were found among 578 patients. Additional screening for CHEK2 mutations in 149 families with familial prostate cancer revealed 11 mutations (5 unique) in nine families. These mutations included two frameshift and three missense mutations. Importantly, 16 of 18 unique CHEK2 mutations identified in both sporadic and familial cases were not detected among 423 unaffected men, suggesting a pathological effect of CHEK2 mutations in prostate cancer development. Analyses of the two frameshift mutations in Epstein Barr virus-transformed cell lines, using reverse-transcriptase polymerase chain reaction and western blot analysis, revealed abnormal splicing for one mutation and dramatic reduction of CHEK2 protein levels in both cases. Overall, our data suggest that mutations in CHEK2 may contribute to prostate cancer risk and that the DNA-damage-signaling pathway may play an important role in the development of prostate cancer. PMID- 12533789 TI - Genomic screen for genes involved in mammalian craniofacial development. AB - Using a subtractive hybridisation approach, we enriched for genes likely to play a role in embryonic development of the mammalian face and other structures. This was achieved by subtracting cDNA derived from adult mouse liver from that derived from 10.5 dpc mouse embryonic branchial arches 1 and 2. Random sequencing of clones from the resultant library revealed that a high percentage correspond to genes with a previously established role in embryonic development and disease, while 15% represent novel or uncharacterised genes. Whole mount in situ hybridisation analysis of novel genes revealed that approximately 50% have restricted expression during embryonic development. In addition to expression in branchial arches, these genes showed a range of expression domains commonly including neural tube and somites. Notably, all genes analysed were found to be expressed not only in the branchial arches but also in the developing limb buds, providing support for the hypothesis that development of the limbs and face is likely to involve analogous molecular processes. PMID- 12533790 TI - Resistance of IAPs to methylation reprogramming may provide a mechanism for epigenetic inheritance in the mouse. AB - Genome-wide epigenetic reprogramming by demethylation occurs in early mouse embryos and primordial germ cells. In early embryos many single-copy sequences become demethylated both by active and passive demethylation, whereas imprinted gene methylation remains unaffected. In primordial germ cells single-copy and imprinted sequences are demethylated, presumably by active demethylation. Here we investigated systematically by bisulphite sequencing the methylation profiles of IAP and Line1 repeated sequence families during preimplantation and primordial germ cell development. Whereas Line1 elements were substantially demethylated during both developmental periods, IAP elements were largely resistant to demethylation, particularly during preimplantation development. This may be desirable in order to prevent IAP retrotransposition, which could cause mutations. In turn, this can result in the transgenerational inheritance of epigenetic states of IAPs, which could lead to heritable epimutations of neighbouring genes through influencing their transcriptional states. PMID- 12533791 TI - Odontoblast-specific expression of cre recombinase successfully deletes gene segments flanked by loxP sites in mouse teeth. AB - Embryonic or neonatal lethality of mice with targeted disruption of critical genes preclude them from further characterization of specific roles of these genes during postnatal development and aging. In order to study the molecular roles of such genes in teeth, we generated transgenic mouse lines expressing bacteriophage Cre recombinase under the control of the mouse dentin sialophosphoprotein (dspp) gene promoter. The expression of Cre recombinase protein was mainly detected in the nucleus of the odontoblasts. The efficiency of Cre activity was analyzed by crossing the Dspp-Cre mice with ROSA26 reporter (R26R) mice. The offspring with both genotypes have shown specific deletion of intervening sequences flanked by loxP sites upstream of the reporter gene, thereby facilitating the expression of the beta-galactosidase (beta-gal) gene in the teeth. The activity of beta-gal was initially observed in the odontoblasts of 1-day-old mice and increased with tooth development. Almost all of the odontoblasts have shown lacZ activity by 3 weeks of age. We could not detect Cre recombinase activity in any other cells, including ameloblasts. These studies indicate that the Dspp-Cre transgenic mice will be valuable to generate odontoblast-specific gene knockout mice so as to gain insight into the molecular roles of critical genes in the odontoblasts during dentinogenesis. PMID- 12533793 TI - Expression of scFv antibodies in Xenopus embryos to disrupt protein function: implications for large-scale evaluation of the embryonic proteome. AB - SUMMARY: We evaluated the use of single-chain antibody (scFv) expression as a tool to disrupt the function of specific proteins in embryos of the frog, Xenopus laevis. The expression of scFvs that recognize the bone morphogenetic protein receptor (ALK3) or the fibroblast growth factor receptor1 (FGFR1) as endoplasmic reticulum-anchored proteins caused distinct developmental defects that were virtually indistinguishable from the defects caused by expression of the dominant negative forms of each receptor. These results demonstrate that scFvs from phage display libraries can be readily fashioned into effective and specific inhibitors of signaling pathways in developing embryos. In addition, as several effective scFvs against a specific target can be isolated rapidly, this approach represents a valuable new tool for large-scale functional analysis of the embryonic proteome. PMID- 12533794 TI - Polycomb group proteins ESC and E(Z) are present in multiple distinct complexes that undergo dynamic changes during development. AB - The Polycomb Group proteins are required for stable long-term maintenance of transcriptionally repressed states. Two distinct Polycomb Group complexes have been identified, a 2-MDa PRC1 complex and a 600-kDa complex containing the ESC and E(Z) proteins together with the histone deacetylase RPD3 and the histone binding protein p55. We report here that there are at least two embryonic ESC/E(Z) complexes that undergo dynamic changes during development and a third larval E(Z) complex that forms after disappearance of ESC. We have identified a larger embryonic ESC complex containing RPD3 and p55, along with E(Z), that is present only until mid-embryogenesis, while the previously identified 600-kDa ESC/E(Z) complex persists until the end of embryogenesis. Constitutive overexpression of ESC does not promote abnormal persistence of the larger or smaller embryonic complexes and does not delay a dissociation of E(Z) from the smaller ESC complex or delay appearance of the larval E(Z) complex, indicating that these changes are developmentally programmed and not regulated by the temporal profile of ESC itself. Genetic removal of ESC prevents appearance of E(Z) in the smaller embryonic complex, but does not appear to affect formation of the large embryonic ESC complex or the PRC1 complex. We also show that the ESC complex is already bound to chromosomes in preblastoderm embryos and present genetic evidence that ESC is required during this very early period. PMID- 12533792 TI - Expression of conditional cre recombinase in epithelial tissues of transgenic mice. AB - SUMMARY: Keratin 18 (K18) expression is a defining characteristic of internal epithelial cells of mammals. Here, we used the K18 gene and an internal ribosome entry site (IRES) to express green fluorescent protein, human placental alkaline phosphatase, and a modified Cre recombinase in an epithelial specific pattern in transgenic mice. The K18-driven alkaline phosphatase was expressed in liver, kidney, uterine endometrium, and other internal epithelia. The enzymatic activity of the Cre recombinase-mutant estrogen receptor fusion protein was dependent on tamoxifen administration and resulted in a mosaic pattern in internal epithelia, including bladder, uterus, liver, and kidney. This conditional Cre activity in internal epithelial organs should be valuable for strategies utilizing Cre for activation of gene expression. This study demonstrates that the tissue-specific, position-independent transcriptional activity of the K18 gene is not compromised by the use of an IRES element for the expression of a second protein from a bicistronic mRNA. PMID- 12533795 TI - Oleosin expression and trafficking during oil body biogenesis in tobacco leaf cells. AB - We have established a versatile method for studying the interaction of the oleosin gene product with oil bodies during oil body biogenesis in plants. Our approach has been to transiently express a green fluorescent protein (GFP)-tagged Arabidopsis oleosin gene fusion in tobacco leaf cells containing bona fide oil bodies and then to monitor oleosin-GFP expression using real-time confocal laser scanning microscopy. We show that normally non-oil-storing tobacco leaf cells are able to synthesize and then transport oleosin-GFP fusion protein to leaf oil bodies. Synthesis and transport of oleosin-GFP fusion protein to oil bodies occurred within the first 6 h posttransformation. Oleosin-GFP fusion protein exclusively associated with the endoplasmic reticulum and was trafficked in a Golgi-independent manner at speeds approaching 0.5 microm sec(-1) along highly dynamic endoplasmic reticulum positioned over essentially static polygonal cortical endoplasmic reticulum. Our data indicate that oil body biogenesis can occur outside of the embryo and that oleosin-GFP can be used to monitor early events in oil body biogenesis in real-time. PMID- 12533796 TI - Overlapping deletions define novel embryonic lethal loci in the mouse t complex. AB - SUMMARY: The t complex region of mouse chromosome 17 contains genetic information critical for embryonic development. To identify and map loci required for normal embryogenesis, a set of overlapping deletions (D17Aus9(df10J), D17Aus9(df12J), and D17Aus9(df13J)) surrounding the D17Aus9 locus and one encompassing the T locus, Del(17)T(7J), were bred in various combinations and the consequences of nullizygosity in overlapping regions were examined. The results indicated that there are at least two functional units within 1 cM of D17Aus9. l17J1 is a peri implantation lethal mutation within the region deleted in D17Aus9(df13J), whereas l17J2 is a later-acting lethal defined by the region of overlap between Del(17)T(7J) and D17Aus9(df12J). Del(17)T(7J)/D17Aus9(df12J) embryos die around 10.5 dpc. The development of the mutant embryos is characterized by lack of axial rotation, an abnormal notochord structure, and a ballooning pericardium. These studies demonstrate the value of overlapping deletion complexes, as opposed to individual deletion complexes, for the identification, mapping, and analysis of genes required for embryonic development. PMID- 12533798 TI - Analyzing the evolution of the opioid/orphanin gene family. AB - Advances in molecular biology have made it possible to rapidly obtain the amino acid sequence of neuropeptide precursors-either by cloning and sequencing the cDNA that encodes the precursor, or by reconstructing the arrangement of exons and introns in a neuropeptide-coding gene through genomic approaches. The databases generated from these molecular approaches have been used to design probes to identify the cells that express the gene, or to ascertain the rate of expression of the gene, and even to predict the post-translational modifications that can generate functional neuropeptides from a biologically inert precursor. Although the power of these approaches is substantial, it is appreciated that a gene sequence or an mRNA sequence reflects the potential products that may be assembled in a secretory cell. To understand the functional capabilities of the secretory cell, the molecular genetics approaches must be combined with procedures that actually characterize the end-products generated by the secretory cell. Recent advances in two-dimensional gel electrophoresis and mass spectrometry now make it possible to analyze neuropeptides from a relatively small amount of tissue. These procedures can reveal novel end-products, tissue specific endoproteolytic cleavage events, and developmental shifts in post translational processing schemes. A gene family that illustrates all of these processes and the advantages of combining genomics with proteomics is the opioid/orphanin gene family. PMID- 12533799 TI - Proteome analysis in the study of the bacterial heat-shock response. AB - In recent years, it has become clear that, in addition to the regulation of the expression of specific genes, there are global regulatory systems that control the simultaneous expression of a large number of genes in response to a variety of environmental stresses. The first of these global control systems, and of substantial importance, is the heat-shock response. The heat-shock response is characterized by the induction of a large set of proteins (heat-shock proteins HSPs) upon shifts to higher temperature and upon exposure to conditions in which proteins are denatured (i.e., alcohols, heavy metals). The heat-shock response is universal and many of the heat-shock proteins are highly conserved among species. In bacteria, the heat-shock response has been studied extensively in several Gram positive bacteria (Bacillus subtilis) and in the Gram-negative bacteria (i.e., Escherichia coli, Agrobacterium tumefaciens). The first recognition of the molecular abundance of the bacterial heat-shock proteins took place with the introduction of high-resolution two-dimensional polyacrylamide gels (2D gels) to analyze complex mixtures of cellular proteins. Two-dimensional gels, followed by mass spectrometry, were used to define the heat-shock stimulons in several bacteria, and to study the regulatory elements that control the heat-shock response. Here, we review the heat-shock response and its regulation in bacteria. The review will emphasize the use of proteome analysis in the study of this response, and will point out those open questions that can be investigated with proteomics, including mass spectrometry techniques. PMID- 12533800 TI - Molecular constituents and phosphorylation-dependent regulation of the post synaptic density. AB - The post-synaptic density (PSD) contains receptors with associated signaling- and scaffolding-proteins that organize signal-transduction pathways near the post synaptic membrane. The PSD plays an important role in synaptic plasticity, and protein phosphorylation is critical to the regulation of PSD function, including learning and memory. Recently, studies have investigated the protein constituents of the PSD and substrate proteins for various protein kinases by proteomic analysis. The present review focuses on the molecular properties of PSD proteins, and substrates of protein kinases and their regulation by phosphorylation in order to understand the role of PSD in synaptic plasticity. PMID- 12533801 TI - Modern strategies for protein quantification in proteome analysis: advantages and limitations. AB - Over the last 3 years, a number of mass spectrometry-based methods for the simultaneous identification and quantification of individual proteins within complex mixtures have been reported. Most, if not all, of such strategies apply a two-step approach: the first for the separation of proteins or peptides, and the second uses mass spectrometry to identify and quantify the individual components. To simplify the outcome of both steps, certain chemicals and heavy-isotope labeling are commonly used in the early stages of sample preparation (except in differential fluorescence labeling protocols). The ultimate goal of these strategies is to be able to identify every protein expressed in a cell or tissue, and to determine each protein's abundance, state of modification, and possible involvement in multi-protein complexes. In this review, an attempt is made to highlight the salient characteristics of the existing strategies with particular attention to their strengths and weaknesses. PMID- 12533803 TI - Detection of maternal serum hCG glycoform variants in the second trimester of pregnancies affected by Down syndrome using a lectin immunoassay. AB - AIM: To assess whether glycoform variants of human chorionic gonadotrophin (hCG) are present in altered concentrations in the maternal serum in pregnancies affected by Down syndrome. METHODS: In a series of 50 cases of pregnancies complicated by Down syndrome and 278 unaffected pregnancies, we have examined maternal serum levels of hCG glycoforms (GlyhCG) in samples collected in the second trimester (14 to 21 weeks) using a sialic acid binding lectin immunoassay. We have compared these levels with those of other second trimester serum markers (Free beta-hCG, alpha fetaprotein (AFP) and Total hCG) and modelled detection rates and false positive rates of various biochemical markers in conjunction with maternal age using a maternal age standardized population. RESULTS: Maternal serum GlyhCG in cases of Down syndrome was significantly elevated (Median MoM 1.81) with 15 of 50 (30%) cases above the 95th centile for unaffected pregnancies. Free beta-hCG was also elevated (Median MoM 2.16) with 18 of 50 (36%) cases above the 95th centile. AFP levels were reduced (Median MoM 0.75) with 9 of 50 (18%) cases below the 5th centile. Total hCG levels whilst elevated (Median MoM 1.88) had only 15 of 50 (30%) cases above the 95th centile. Maternal serum GlyhCG levels showed significant correlation with total hCG and free beta hCG (r = 0.6880 and 0.6922) in the Down group but not with AFP (r = 0.1237). When GlyhCG was combined together with AFP and maternal age, at a 5% false positive rate, the modelled detection rate was 53%, some 13% lower than when free beta-hCG was used and some 7% lower than when total hCG was used. CONCLUSION: Maternal serum GlyhCG, as measured by the sialic acid-binding lectin immunoassay is unlikely to be of additional value when screening for Down syndrome in the second trimester. PMID- 12533804 TI - Prenatal diagnosis of molybdenum cofactor deficiency and isolated sulfite oxidase deficiency. AB - Molybdenum cofactor deficiency and isolated sulfite oxidase deficiency are autosomal recessive inborn errors of metabolism with severe neurological symptoms resulting from a lack of sulfite oxidase activity. The deficiencies can be diagnosed prenatally by monitoring sulfite oxidase activity in chorionic villus sampling (CVS) tissue. In those families in which the specific defects have been identified, diagnosis can be achieved by mutation analysis or linkage studies directed at affected genes. These include MOCS1, MOCS2 or GEPH, in cases of molybdenum cofactor deficiency, or SUOX in patients with isolated sulfite oxidase deficiency. PMID- 12533805 TI - Assessment of the thymus at echocardiography in fetuses at risk for 22q11.2 deletion. AB - OBJECTIVES: An absent or hypoplastic thymus is common in patients with 22q11.2 deletion (del22q11.2). We sought to determine whether fetal echocardiography could identify absence of the thymus as a diagnostic tool in pregnancies at risk for fetal del22q11.2. METHODS: We evaluated the fetal thymus in 16 consecutive pregnancies at risk for fetal del22q11. Fourteen of the fetuses had a conotruncal cardiac lesion, one had a twin with a conotruncal lesion, and in one the mother had a diagnosis of del22q11.2. The fetal thymus assessment was performed by an individual who was not aware of the del22q11.2 status of the fetus. RESULTS: By 2D imaging, the thymus was identified in the anterosuperior mediastinum as a subtle hypoechogenic area. In nine cases, the thymus was demonstrated prenatally and none had del22q11.2. However, in one case the thymus was only seen on follow up fetal echocardiography. In six cases, the thymus could not be identified and all six had del22q11.2. In one additional case, analyzed retrospectively, the thymus could not be assessed. The status of the thymus was confirmed on postnatal echocardiography or autopsy in 11 of the 15 cases assessed prenatally. CONCLUSIONS: Our study suggests that fetal echocardiography can assess the thymus in most cases at risk for del22q11.2. This information may be useful in counseling women/couples who decline amniocentesis or who are awaiting amniocentesis results. PMID- 12533806 TI - Triploidy in a twin pregnancy: small placenta volume as an early sonographical marker. AB - INTRODUCTION: We report a case of a twin pregnancy with triploidy of maternal phenotype of one foetus and no chromosomal anomaly of the other twin and the role of sonographical placental volumetry. CASE: At 12 weeks of gestation, a dichorionic twin pregnancy discordant in growth is diagnosed. 3D ultrasound reveals a distinctly small placental volume of foetus II. Amniocentesis at 16 weeks discloses triploidy of this foetus. Sonography reveals asymmetrical foetal growth retardation, a severe heart defect and bilateral cleft lip and palate, typical findings in triploidy. Selective feticide at week 20+3 is followed by pre term birth of foetus I at 27 weeks. CONCLUSION: Small placental volume in addition to growth restriction of one foetus early in the course of a twin pregnancy could be an important early marker influencing the decision for chorionic villous sampling at 12 weeks instead of amniocentesis at 16 weeks and it could lead to an earlier selective pregnancy termination of a triploid twin. This would lower the risk of pre-term birth and enable a better outcome for the remaining healthy foetus. PMID- 12533807 TI - Second trimester diagnosis of Neu Laxova syndrome. AB - This is the first report of a prenatally diagnosed case of Neu Laxova syndrome (NLS) from India. This also includes a case of NLS in monochorionic diamniotic twins and two more cases in which we were able to detect most of the features of NLS as early as 19 to 20 weeks by routine antenatal ultrasonography. Severe intrauterine growth retardation (IUGR), microcephaly, central nervous system (CNS) abnormality, joint contractures, and abnormal facies are the major diagnostic features observed in prenatal ultrasonography. Risk factors such as consanguinity and history of intrauterine death or stillbirth in siblings have been noted in all the cases, but none of the three families that were reported had previously had an affected fetus. The spectrum of skin manifestations and frequency of occurrence of major clinical features of the syndrome have been discussed. Review of the literature on NLS and possibility of detecting the syndrome in the second trimester is discussed. PMID- 12533808 TI - Congenital erythropoietic porphyria (Gunther's disease): two cases with very early prenatal manifestation and cystic hygroma. AB - Congenital erythropoietic porphyria (CEP) or Gunther's disease is the rarest form of the porphyrias. The disease is usually diagnosed at birth or during early infancy, but rarely in utero. We describe here the first two cases of very early prenatal expression of CEP with cystic hygroma diagnosed at 14 weeks in the first fetus and at 19 weeks in the second. Both fetuses presented with severe nonimmune hydrops fetalis as early as 19 and 22 weeks, associated with intrauterine growth retardation, hyperechogenic kidneys and bones. Amniotic fluid was dark brown and uro- and coproporphyrin I was dramatically increased. Molecular screening of the CEP gene detected heterozygous C73R mutation in both fetuses, the other parental mutation being as yet unknown. PMID- 12533809 TI - Prenatal exclusion of Leigh syndrome due to T8993C mutation in the mitochondrial DNA. AB - Leigh syndrome (LS) is a mitochondrial encephalopathy that is caused by a mutation either in the mitochondrial DNA (mtDNA) or in the nuclear encoded genes of the mitochondrial proteins. Prenatal diagnosis of defects in the mtDNA is usually problematic because of mtDNA heteroplasmy and tissue specificity. However, the mutations T8993 G/C in the ATP synthase subunit 6 gene of the mtDNA show a more even tissue distribution and do not appear to change significantly over time. There are only few reports of prenatal diagnosis of the T8993G mutation in Leigh disease. Here we describe the first prenatal genetic testing of T8993C in a fetus of a mother whose previous child had died of Leigh syndrome due to the T8993C mutation. Mutant load in the chorionic villus sample (CVS) as well as in amniocytes was undetectable, thus predicting a very high likelihood of an unaffected outcome, indicative of a healthy baby. The diagnosis was confirmed after birth. Gathering data on the prenatal diagnosis of mtDNA mutations is of great importance so that prenatal diagnosis of both T8993G and T8993C mutations can be offered routinely. PMID- 12533810 TI - HLA-G positive trophoblastic cells in transcervical samples and their isolation and analysis by laser microdissection and QF-PCR. AB - OBJECTIVE: To assess the frequency of cytotrophoblastic cells in endocervical samples collected by lavage at early stages of gestation using a specific anti HLA-G McAb (G233). From a set of four selected samples, cells identified by immunostaining were collected by laser microdissection and then tested by quantitative fluorescent polymerase chain reaction (QF-PCR) for the presence of paternally derived DNA markers, in order to establish their fetal origin. METHODS: Syncytial fragments and cytotrophoblastic cells from 23 transcervical samples were identified by immunostaining with McAb G233 reacting against HLA-G antigen and with antibodies against cytokeratin. Slides from the same samples were also tested by fluorescent in situ hybridization (FISH), while selected samples were analysed by QF-PCR. Slides from four samples retrieved from mothers with male fetuses were immunolabelled and then cytotrophoblastic cells, syncytial fragments and maternal epithelial cells were collected by laser microdissection and tested by QF-PCR. RESULTS: All endocervical samples retrieved from mothers with male fetuses were found to contain some cells with chromosome Y-specific signals when tested by FISH. Using McAb anti- HLA-G, cytotrophoblastic cellular elements were detected in about 50% of the samples. From four samples, cellular elements identified by immunostaining as cytotrophoblast or syncytial fragments were collected by laser microdissection and shown to be of fetal origin when tested by QF-PCR for the presence of fetal DNA markers. CONCLUSIONS: These results confirm that, during an early phase of gestation, fetal cells are released in the lower uterine cavity and that they can be isolated and analysed for prenatal diagnosis of single gene defects and aneuploidies. PMID- 12533811 TI - Prenatal diagnosis of mosaic ring chromosome 22 associated with cardiovascular abnormalities and intrauterine growth restriction. AB - OBJECTIVES: To present the prenatal diagnosis and perinatal findings of mosaic ring chromosome 22. CASE: Amniocentesis was performed at 18 gestational weeks because of an advanced maternal age. Cytogenetic analysis of the cultured amniotic fluid cells revealed mosaicism for ring chromosome 22, 45,XX, 22[6]/46,XX,r(22)(p13q13.31)[15]. Abnormal fetal sonographic findings included small for gestational age, a ventricular septal defect, and truncus arteriosus. The pregnancy was terminated. Additional phenotypic findings included hypertelorism, epicanthal folds, and abnormal ears. Cytogenetic analysis of the cord blood lymphocytes revealed a complex mosaic karyotype, 45,XX, 22[7]/46,XX,r(22)(p13q13.31)[82]/46,XX,idic r(22)(p13q13.31;p13q13.31)[11]. Cytogenetic analysis of the hepatocytes also revealed mosaic r(22) with mosaicism for idic r(22) and monosomy 22. The deletion of distal 22q and the duplication of 22q11.2 on idic r(22), and the distal 22q deletion on r(22) were demonstrated by fluorescent in situ hybridization (FISH) analysis using 22q terminal probes at 22q13 and a DiGeorge syndrome critical region probe at 22q11.2. The breakpoint on distal 22q13 and the extent of the duplication of 22q on idic r(22) was determined by examining polymorphic markers specific for chromosome 22 using quantitative fluorescent polymerase chain reaction assays. The chromosomal aberration was of maternal origin. CONCLUSION: Molecular and FISH studies allow a better delineation of some prenatally detected aneuploidy syndromes and help elucidate the genetic pathogenesis. Fetuses having mosaic r(22) with a low level mosaicism for r(22) duplication/deletion may present cardiovascular abnormalities and intrauterine growth restriction on prenatal ultrasound. PMID- 12533812 TI - Fetal liver hyperechogenicity on sonography may be a serendipitous sign of a transient myeloproliferating disorder. AB - OBJECTIVE: To demonstrate that thorough antenatal and postnatal investigations are helpful in any genetic syndrome. CASE REPORT: A woman without a specific risk had a finding of hyperechogenic foci at ultrasound. Karyotype on fetal blood was 47, XY+21 and extensive biochemical investigation revealed abnormal liver function. Given the suspicion of a coincidental hepatic compromise, targeted postmortem examination was performed, which showed hepatic fibrosis, megakaryocytes proliferation and invasion of sinusoid capillaries, all suggestive of a transient myeloproliferative disorder. CONCLUSION: When compared with chorionic villous and amniotic fluid sampling, fetal blood allows the collection of additional data, which may help in understanding the pathological process behind an apparently serendipitous association. PMID- 12533813 TI - Congenital nephropathy and ventriculomegaly: a report of four cases. AB - Congenital nephrotic syndrome with ventriculomegaly and a normal karyotype is a rare association. We report four cases, three of which were conceived consecutively by one couple. All the cases were associated with elevated maternal serum alpha-fetoprotein. Renal histology in one fetus demonstrated colloid filled cysts distributed in the corticomedullary area. Transmission electron microscopy of the glomeruli showed normally developed foot processes and confirmatory genetic studies excluded Finnish congenital nephrotic syndrome. It is probable that congenital nephropathy in conjunction with ventriculomegaly is the result of an autosomal recessive syndrome. PMID- 12533814 TI - Detection of gamma-globin mRNA in fetal nucleated red blood cells by PNA fluorescence in situ hybridization. AB - OBJECTIVES: Fetal nucleated red blood cells (NRBC) that enter the peripheral blood of the mother are suitable for non-invasive prenatal diagnosis. The application of peptide nucleic acid (PNA) probes for tyramide amplified flow fluorescence in situ hybridization (FISH) detection of gamma-globin mRNA in fixed fetal NRBC is investigated. METHODS: Hemin-induced K562 cells or nucleated blood cells (NBC) from male cord blood were mixed with NBC from non-pregnant women and analysed using both slide and flow FISH protocols. Post-chorionic villus sampling (CVS) blood samples from pregnant females carrying male fetuses were flow-sorted (2 x 10(6) NBC/sample). Y chromosome-specific PNA FISH was used to confirm that the identified gamma-globin mRNA stained cells were of fetal origin. RESULTS: Flow FISH isolated gamma-globin mRNA positive NBCs showing characteristic cytoplasmic staining were all Y positive. The amplification system generated a population of false positive cells that were, however, easy to distinguish from the NRBCs in the microscope. CONCLUSION: The gamma-globin mRNA specific PNA probes can be used for detection and isolation of fetal NRBCs from maternal blood. The method has additional potential for the study of gamma-globin mRNA levels or the frequency of adult NRBC (F cells) in patients with hemoglobinopathies. PMID- 12533815 TI - The variables affecting nuchal skin-fold thickness in mid-trimester. AB - OBJECTIVES: To find out variables that have an effect on the measured values of nuchal skin-fold thickness (NT) and to formulate a regression equation on the basis of those variables. METHODS: The data on gestational age (GA), cephalic index (CI), presentation (Pr, vertex or breech), and the presence or absence of nuchal cord (NC) were collected prospectively on 548 normal singleton fetuses between 16 and 24 weeks gestation, and independent correlation of those variables with NT was calculated by multiple regression analysis and a regression equation was produced. RESULTS: GA had positive correlation and CI had negative correlation with NT. The nuchal skin-fold was thicker among fetuses with breech presentation rather than those with vertex presentation and increased in the presence of nuchal cord. We calculated the expected NT through these observations: for fetuses presenting vertex, NT = 5.608 + 0.243GA - 0.066CI + NC* and for breech, NT = 2.803 + 0.392GA - 0.066CI + NC*. CONCLUSIONS: This is the first report, which takes GA, CI, Pr, and NC for correlation factors with NT as a whole. These equations and further studies on determining the cutoff values of the nuchal skin-fold thickness may improve the sensitivity and specificity for the detection of Down syndrome and enable to minimize unnecessary invasive procedures such as amniocentesis or cordocentesis. PMID- 12533816 TI - Pitfalls in prenatal diagnosis: cytogenetic analysis in amniocytes fails to detect mosaic r(12). AB - OBJECTIVE: To review the accuracy of a prenatal diagnosis of a missed chromosomal mosaicism in amniotic fluid cell cultures and to see whether adapting the Dutch guidelines would have made any difference to the outcome in this case. METHOD: Metaphases, obtained from cultured amniocytes and peripheral blood lymphocytes, were analyzed with different results. The amniocyte cultures were then reanalyzed and the risk of missing this mosaicism in prenatal analysis was assessed. RESULTS: The prenatal tests performed according to the Dutch guidelines showed a normal female karyotype, but more extensive postnatal analysis revealed a ring chromosome in 50% of the child's lymphocytes. Reanalysis of the original amniocytes confirmed the normal diagnosis, but when more cells from the same and other colonies were analyzed, the ring chromosome was detected. CONCLUSION: The chance of missing such a supernumerary ring mosaicism is very low (about 2% in our case). Given its very rare occurrence and the low chance of it being missed if the existing Dutch guidelines are followed, adapting the number of cells or colonies to be examined for all prenatal diagnoses does not appear to be justified. PMID- 12533817 TI - A simple and sensitive erythroblast scoring system to identify fetal cells in maternal blood. AB - OBJECTIVES: Nucleated red blood cells (NRBCs) of fetal origin appear to have distinguishable characteristics from that of maternal NRBCs in both nuclear morphology and properties of hemoglobin staining. However, these differences have yet to be quantified. Our aim was to develop an erythroblast scoring system using four distinct phenotypic parameters (nuclear roundness, nuclear morphology, gamma hemoglobin staining intensity, and peripheral brightness of the stained cytoplasm) to address this issue. METHODS: NRBCs were isolated from four maternal blood samples by density gradient separation, CD15/45 depletion, and gamma hemoglobin positive selection after elective termination of a trisomy 21 male fetus (47,XY,+21). All cells were deposited onto microscope slides and every NRBC was analyzed according to the scoring system. Each of the four individual parameters was given a value from 0 to 3 points and a combined score was obtained for each cell (range 0-12). Fluorescence in situ hybridization (FISH) using X- and Y-specific probes was performed to determine, on the basis of interphase karyotype, whether the cell was maternal or fetal. RESULTS: The majority of maternal NRBCs were found to have a combined score of 6 or less (103/117) and the majority of fetal NRBCs were found to have a score of 7 or greater (43/53). The proportion of cells that were identified correctly as fetal increased with each ascending category of combined score. For example, 5.7% of NRBCs with a combined score of 5 points or less were found to be fetal, whereas 19.2% of NRBCs with a combined score of 6 points were fetal. At combined scores of 11 and 12 points, 100% of NRBCs were found to be fetal. CONCLUSION: Fetal NRBCs have characteristic morphology and a gamma hemoglobin staining appearance that makes them distinguishable from maternal NRBCs. The scoring system presented here is a simple and sensitive method to distinguish fetal NRBCs from adult cells in maternal blood. This system may have clinical utility for noninvasive prenatal diagnosis as well as applications for basic research into the developmental biology of NRBCs. In addition, these defined parameters may serve as computational classifiers for the automated detection of fetal cells in maternal blood. PMID- 12533819 TI - Diagnosis of cardiac defects: where we have been, where we are, and where we are going. PMID- 12533818 TI - The clinical application of spectral karyotyping (SKY) in the analysis of prenatally diagnosed extra structurally abnormal chromosomes (ESACs). AB - OBJECTIVE: The prenatal detection of de novo extra structurally abnormal chromosomes (ESACs) presents a challenge because the associated risk for congenital anomaly ranges from 100% to practically none, depending on the chromosomal origin. Despite the use of standard cytogenetic techniques and even fluorescence in situ hybridization (FISH), the origin of some ESACs often remains elusive. Spectral karyotyping (SKY) is a molecular cytogenetic technique based on the simultaneous analysis of all chromosomes using a unique probe mix that allows the rapid identification of all chromosomes in 24 colors. The purpose of this study was to evaluate the use of SKY in the characterization of prenatally diagnosed de novo ESACs. METHODS: This series includes five cases of de novo ESACs detected prenatally in routine amniocentesis samples performed for advanced maternal age. Cases of inherited ESACs or ESACs defined by standard cytogenetic techniques were excluded. RESULTS: SKY analysis yielded valuable information, particularly in cases of nonsatellited ESACs: a der(18) and a ring(Y). In a case of a unisatellited der(15), SKY corroborated data obtained by standard cytogenetic techniques and FISH. Finally, in two cases of small bisatellited chromosomes, SKY was noncontributory. CONCLUSIONS: While SKY may be a valuable tool in some cases, especially nonsatellited and ring ESACs, it does have limitations and should be used judiciously in conjunction with other cytogenetic techniques. PMID- 12533820 TI - Detection of mosaic isochromosome 20q in amniotic fluid in a pregnancy with fetal arthrogryposis multiplex congenita and normal karyotype in fetal blood and postnatal samples of placenta, skin, and liver. PMID- 12533821 TI - Trisomy 5p in a prenatal case with ultrasound abnormalities. PMID- 12533822 TI - Chronology of reported denaturing high performance liquid chromatography (DHPLC) based prenatal diagnoses. PMID- 12533823 TI - Prenatal management of mosaic tetrasomy 5p. PMID- 12533824 TI - Colorectal tumourigenesis in carriers of the APC I1307K variant: lone gunman or conspiracy? AB - The APC I1307K allele is believed to predispose to multiple colorectal tumours because the change at the nucleotide level-A(3)TA(4) to A(8)-creates a hypermutable site. Slippage of the A(8) tract undoubtedly occurs more often than expected in the tumours of I1307K carriers, but many of these tumours do not harbour changes at this site. Outside the A(8) tract, the tumours of I1307K carriers appear to harbour fewer somatic frameshift mutations than expected. There is inconsistent evidence as to whether or not I1307K confers an increased risk of colorectal cancer. Most I1307K patients and families who have undergone analysis of somatic APC mutations have been highly selected. It is therefore possible that many APC I1307K carriers with multiple adenomas have a susceptibility to tumours additional to that resulting from the A(8) tract. PMID- 12533825 TI - Epstein-Barr virus-associated carcinomas: facts and fiction. AB - The Epstein-Barr virus (EBV) is associated with several human tumours including lymphoid and epithelial malignancies. Most EBV-associated tumours are rare or occur at higher incidence only in certain geographical regions. The recently reported detection of EBV in gastric, breast, and hepatocellular carcinomas raises the possibility of involvement of the virus in the pathogenesis of common cancers. This article reviews the evidence linking EBV infection to epithelial tumours. It is concluded that at present, there is no convincing evidence to suggest that breast carcinoma and hepatocellular carcinoma are EBV-associated tumours. PMID- 12533826 TI - The characterization of somatic APC mutations in colonic adenomas and carcinomas in Ashkenazi Jews with the APC I1307K variant using linkage disequilibrium. AB - Mutations of the adenomatous polyposis coli (APC) gene play a critical role in the development of colorectal neoplasms. A novel mechanism involves a germline variant, at codon 1307 of the APC gene. The mutation is thought to create an unstable segment of DNA, which facilitates the development of somatic mutations. I1307K has been shown to be ancestral in Middle Eastern populations. The aim of the present study was to confirm this observation for a Western population and to utilize this information in the characterization of the somatic changes in colorectal neoplasia in carriers of I1307K. DNA from 182 US Ashkenazim was screened for the I1307K variant, which was found in 22 (12.1%), and the ancestral nature of this variant was confirmed in this population by showing that all of those with the I1307K variant carried a specific allele at the D5S346 locus, while the majority shared a D5S1385 allele. Subsequently, 79 neoplasms were analysed from 15 I1307K carriers for loss of heterozygosity (LOH) at the D5S346 locus. LOH was detected in 18 neoplasms (23%). Of these 18, four neoplasms showed loss of the I1307K-associated allele, while 14 neoplasms had retained the I1307K associated allele and, by implication, the I1307K variant. PCR products were also cloned into a plasmid vector to isolate individual APC alleles. For those neoplasms with LOH, 13 of the 18 neoplasms (72%) had a somatic mutation, of which 12 involved the I307K-bearing chromosome. This study is consistent with two previous studies in showing that additional somatic mutations close to codon 1307 are almost always on the I1307K-bearing chromosome, but either allele may demonstrate LOH. Further evidence for the interpretation of the action of I1307K as producing DNA instability is provided by analysing multiple neoplasms from the same person and by showing that these neoplasms have differing patterns of LOH and associated somatic mutations. PMID- 12533827 TI - Comparative genomic hybridization analysis of bilateral hyperplasia of usual type of the breast. AB - Hyperplasia of usual type (HUT) is part of the spectrum of benign proliferative disease in the breast and confers an increased risk of developing invasive cancer. Its role as a putative precursor of invasive ductal carcinomas in the breast is, however, controversial. HUT is occasionally seen bilaterally but it is not clear whether there is a genetic predisposition to the process. This study has analysed 14 cases of bilateral HUT in the breast (28 independent lesions) by comparative genomic hybridization (CGH) analysis to define DNA copy number changes and to investigate any commonality in these genetic alterations. The mean number of alterations seen was 1.6 (46/28) in all lesions, with common losses at chromosomes 1p, 16p, 17q, and 22q. Bilaterally, only five alterations were seen in common across both lesions in both breasts, but no single locus was altered preferentially. These data indicate that a proportion of HUT in the breast are indeed clonal, neoplastic proliferations which exhibit genetic alterations in common with invasive breast cancer, albeit at very low levels. The limited data from these experiments do not reveal a specific predisposition locus. PMID- 12533828 TI - Barrett's adenocarcinomas resemble adenocarcinomas of the gastric cardia in terms of chromosomal copy number changes, but relate to squamous cell carcinomas of the distal oesophagus with respect to the presence of high-level amplifications. AB - Three different cancers predominantly occur at the gastro-oesophageal junction: squamous cell carcinomas of the distal oesophagus, adenocarcinomas of the distal oesophagus (Barrett's carcinomas), and adenocarcinomas of the gastric cardia. The aim of the present study was to investigate how, and to what extent, Barrett's carcinoma differs from adenocarcinoma of the gastric cardia on the one hand and squamous cell carcinoma of the distal oesophagus on the other, with respect to chromosomal aberrations and related gene expression. The present study analysed 14 squamous cell carcinomas, 24 Barrett's carcinomas, and 16 carcinomas of the gastric cardia. Comparative genomic hybridization revealed chromosomal abnormalities in all cases. Typical chromosomal aberrations for the squamous cell carcinoma type were gains at 3q and 11q13, and losses at 3p, 4q, 9p, 11q, and 13q. In contrast, typical copy number changes for both cardiac and Barrett's adenocarcinomas were gains at 2q, 7p, and 13q, and losses at 17p. High-level amplification occurred in all three groups, but its frequency in the cardiac carcinomas was lower than in the other two groups. In conclusion, squamous cell carcinomas are characterized by chromosomal aberrations which are distinct from those seen in carcinomas of the gastric cardia and in Barrett's adenocarcinomas. With respect to Barrett's cancer, the chromosomal aberrations more closely reflect the adenocarcinoma phenotype than the squamous origin of the epithelium. PMID- 12533829 TI - Diverse p53 alterations in ulcerative colitis-associated low-grade dysplasia: full-length gene sequencing in microdissected single crypts. AB - In long-standing ulcerative colitis (UC), p53 mutations have been shown to occur by indirect detection methods such as PCR-SSCP. To clarify whether p53 gene mutations are early events in UC-associated neoplasia and to analyse clonality within dysplasia-associated lesions or masses (DALMs), the entire coding region of the p53 gene was analysed in DNA of microdissected single crypts by the polymerase chain reaction (PCR)-direct sequencing method. With a novel microdissection method using serial histological sections, the p53 gene (exons 2 11) was analysed in a total of 11 regenerative crypts and 76 single crypts within seven DALMs selected from three colectomy specimens of long-standing UC patients. Although p53 point mutations were found in at least one crypt in each DALM, heterogeneity in terms of the presence and the type of genetic change was marked, except in one carcinoma. As early events, p53 gene mutations were apparent even in some regenerative crypts (8/12 crypts). Some were of silent type. Altered p53 protein expression was confirmed in only 14/32 mutated crypts and was also evident in 24 other non-mutated examples by immunostaining of serial sections. Polyclonal p53 gene mutations were found in regenerative (REG) crypts and low grade dysplasia (LGD), but monoclonal changes were noted in high-grade dysplasia (HGD) or carcinoma (Ca) in long-standing UC. At the single crypt level, however, p53 point mutations were not always linked to p53 overexpression, indicating a discrepancy between gene alteration and protein accumulation in LGD. PMID- 12533830 TI - Association of trypsin expression with tumour progression and matrilysin expression in human colorectal cancer. AB - Overexpression of the matrix serine protease (MSP) trypsin has been implicated in tumour growth, invasion, and metastasis. The objective of this study was to clarify the clinicopathological and prognostic significance of trypsin expression in colorectal cancer. This study analysed the association between immunohistochemically detected trypsin expression in colorectal cancer and clinicopathological characteristics, and investigated whether trypsin is a predictor of recurrence and/or survival. Trypsin immunoreactivity was more intense at the invasive front than in the superficial part of the tumour. Sections with immunostaining signals in more than 30% of carcinoma cells at the invasive front, which were observed in 48 cases (48%), were judged to be positive for trypsin. Trypsin positivity was significantly correlated with depth of invasion, lymphatic and venous invasion, lymph node and distant metastasis, advanced pathological tumour-node-metastasis (TNM) stage, and recurrence. Patients with trypsin-positive carcinoma had significantly shorter overall and disease-free survival periods than did those with trypsin-negative carcinoma. Trypsin retained its significant predictive value for overall and disease-free survival in multivariate analysis that included conventional clinicopathological factors. It is well known that trypsin activates matrilysin (matrix metalloproteinase-7), which plays an important role in colorectal cancer progression. Patients with concordant overexpression of trypsin and matrilysin at the invasive front, in which they were often co-localized, had the worst prognosis. Trypsinogen-1-transfected HCT116 colon cancer cells showed not only trypsin activity, but also active matrilysin activity and were more invasive in vitro than mock-transfected HCT116 cells. These results suggest that trypsin plays a key role in the progression of colorectal cancer. Detection of trypsin expression as well as matrilysin is useful for the prediction of recurrence in and poor prognosis of colorectal cancer patients. PMID- 12533831 TI - Significance of aberrant (cytoplasmic/nuclear) expression of beta-catenin in pancreatoblastoma. AB - This study concerns the significance of aberrant (nuclear/cytoplasmic) expression of beta-catenin in pancreatoblastoma (PBL). On immunohistochemistry, all seven PBLs examined showed nuclear/cytoplasmic expression of beta-catenin, predominantly in the squamoid corpuscles (SCs). In areas with acinar/ductular differentiation, few tumour cells displayed nuclear/cytoplasmic expression of beta-catenin and more than half of the tumour cells showed membranous expression. Two out of five (40%) tumours examined showed missense mutations in codons 33 and 37 of exon 3 of the beta-catenin gene. No mutation of the adenomatous polyposis coli (APC) gene was detected in two of the remaining three tumours. Amplifiable DNA for APC analysis was not obtained from the one other tumour. Immunoreactivity for cyclin D1, one of the nuclear targets of beta-catenin, was found predominantly in the SCs of the seven tumours. In contrast, the Ki-67 labelling index was 2-4% (median 3%) in the SCs and 8-18% (median 12%) in the other areas, indicating a negative correlation with nuclear cyclin D1 reactivity. These results imply that in PBLs, nuclear/cytoplasmic accumulation of beta-catenin and overexpression of its target gene cyclin D1 are not associated with the induction of tumour cell proliferation. Nuclear/cytoplasmic accumulation of beta-catenin may be related to the morphogenesis of the SCs that are considered most characteristic for PBL. PMID- 12533832 TI - Liver pathology and hepatocarcinogenesis in a long-term mouse model of erythropoietic protoporphyria. AB - Erythropoietic protoporphyria (EPP) is an inherited disease of haem synthesis caused by a mutation in one of the alleles of the enzyme ferrochelatase. This mutation leads to partial deficiency of the enzyme, resulting in increased concentrations of protoporphyrin (PP) in blood, liver, and faeces. Five to ten per cent of patients with EPP develop severe liver disease characterized by the presence of PP deposits. This study used histochemistry and immunohistochemistry to investigate the histopathological features present in the livers of 44 mice with a heterozygous or homozygous point mutation in the ferrochelatase gene (fch/+ and fch/fch mice, respectively). Some fch/+ mouse livers showed mixed steatosis and large cell dysplasia. The livers of fch/fch mice showed periportal or septal fibrosis accompanied by an atypical ductular reaction. These findings suggest that the obstruction and damage of a proportion of large and small bile ducts by PP deposits cause an accumulation of PP in the parenchyma, which leads to damage and loss of hepatocytes due to the toxic effects of PP. The classical stages of hepatocarcinogenesis were observed and hepatic progenitor cells appear to be involved in this process. PP acts as the promoting agent and is probably also the initiating agent. PMID- 12533833 TI - High-throughput tissue microarray analysis of G1-cyclin alterations in classical Hodgkin's lymphoma indicates overexpression of cyclin E1. AB - Deregulation of G1-cyclins (CCN) plays a key role in the pathogenesis of many human malignancies, including non-Hodgkin's lymphomas (NHLs). In contrast to NHL, little is known about phenotypic and genotypic changes in the regulation of the cell cycle in classical Hodgkin's lymphoma (cHL). To facilitate analysis of aberrant gene expression in cHL, a lymphoma tissue microarray (TMA) containing 752 cores of 330 different cHL samples was constructed. Direct comparison of Epstein-Barr virus (EBV) latent membrane protein 1 (LMP-1) expression in Hodgkin's and Reed-Sternberg (HRS) cells on conventional full sections with the corresponding duplicate/triplicate tumour cores on the TMA showed a concordance of 100%, indicating that cHL-TMA is a reliable and representative method for evaluating gene expression profiles in situ. Using TMA technology, protein expression and gene amplification of different G1-CCNs in cHL were analysed. Among the G1-CCNs analysed, cyclin E (CCNE) was expressed in 212/253 cases (84%). In most of the individual tumours, over 75% of the HRS cells stained positive for CCNE, suggesting that CCNE is overexpressed in cHL. This overexpression was not due to CCNE gene amplification, as judged by fluorescence in situ hybridization, and did not correlate with EBV infection, as assessed by the expression of LMP-1. Thus, the overexpression of CCNE could be caused by profound changes in HRS cell cycle regulation that could contribute to the malignant phenotype. PMID- 12533834 TI - Diversity in expression and prognostic significance of G1/S cyclins in human primary lung carcinomas. AB - Expression of cyclin A, cyclin E and cdk2 was examined immunohistochemically in 144 cases of primary non-small cell lung carcinoma to evaluate their prognostic value. Cyclin A was co-expressed with cdk2 in the proliferating cells, ie those showing positive Ki-67 staining. The labelling index (LI) of cyclin A revealed a positive correlation with the S-phase fraction and an inverse correlation with histological differentiation. Furthermore, high cyclin A LIs indicated a poor prognosis in all histological types. Cyclin E exhibited a characteristic staining pattern: in squamous cell carcinoma (SCC), differentiated cells without Ki-67 staining revealed cyclin E positivity with expression of cdk2. Conversely, in adenocarcinoma (AC), proliferating cells revealed cyclin E positivity. Cases of large cell carcinoma showed heterogeneous cyclin E staining patterns, unlike those of SCC or AC. Statistical analyses also revealed a marked contrast between SCC and AC. In AC, the LI of cyclin E was inversely correlated with histological differentiation and a high LI predicted a worse prognosis. In contrast, in SCC, the LI of cyclin E correlated positively with histological differentiation and better prognosis. However, the expression levels of cyclin E mRNA evaluated by quantitative RT-PCR were higher in poorly differentiated SCC and AC, suggesting that protein turnover plays a large role in determining cyclin E protein levels. Although the expression of cyclins was demonstrated to be diversely regulated depending on the histological type, the combined immunohistochemical analyses performed in this study on these proteins could be useful tools for evaluating patient prognosis in lung carcinomas. PMID- 12533835 TI - Comparison of immunohistochemistry for activated caspase-3 and cleaved cytokeratin 18 with the TUNEL method for quantification of apoptosis in histological sections of PC-3 subcutaneous xenografts. AB - The terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labelling (TUNEL) technique has been extensively used for the detection and quantification of apoptosis in histological tissue sections. However, the interpretation and specificity of this assay have been controversial. With accumulating knowledge of the molecular mechanisms of cell death and the discovery of the caspases as key mediators of apoptosis, more direct and earlier measurements of apoptosis in tissue sections have emerged. This study, using antibodies that specifically recognize activated caspase-3 and caspase-cleaved cytokeratin (CK) 18, evaluated whether immunohistochemical stains would improve the detection and quantification of apoptosis in tissue sections, compared with the TUNEL assay. Tumour xenografts of the prostate cancer cell line PC-3 were used as an example, since these tissues contain large numbers of cells undergoing apoptosis. Apoptotic cells were quantified and apoptotic indices were calculated by computer-assisted image analysis following identification of apoptotic cells by morphological analysis, the TUNEL assay, activated caspase-3 and cleaved CK18 immunohistochemistry. The results indicated that activated caspase-3 immunohistochemistry was an easy, sensitive, and reliable method for detecting and quantifying apoptosis in this model. An excellent correlation (R = 0.89) between the apoptotic indices obtained using activated caspase-3 and cleaved CK18 immunostaining was observed. A good correlation (R = 0.75) between the apoptotic indices obtained using activated caspase-3 immunostaining and the TUNEL assay was also found. Activated caspase-3 immunohistochemistry is therefore recommended for the detection and quantification of apoptosis in tissue sections. PMID- 12533836 TI - Myocytes positive for in situ markers for DNA breaks in human hearts which are hypertrophic, but neither failed nor dilated: a manifestation of cardiac hypertrophy rather than failure. AB - The significance of DNA breaks reported in failing hearts is controversial, although they may suggest myocyte apoptosis and may thus be responsible for the progression of heart failure. This study attempted to check the validity of the in situ markers for DNA breaks for detecting myocyte death and to evaluate separately two factors, failure or hypertrophy, crucial for DNA breaks in pathological human hearts. In the autopsy study, myocytes showed positivity for in situ nick end-labelling (TUNEL) and of Taq and Pfu polymerase-based in situ ligation assays not only in dilated cardiomyopathy (DCM, n = 9) with failure, but also in hypertrophic cardiomyopathy (HCM, n = 8) and hypertensive heart disease (HHD, n = 4) without failure. There was a significant correlation between each in situ marker and heart weight. The incidence of TUNEL-positive myocytes always exceeded that seen in in situ ligation assays. In addition, there were significant correlations between the in situ markers and the expression of the proliferating cell nuclear antigen (PCNA) and of the spliceosome component of 35 kD (SC-35). Similarly, in the left ventricular biopsy study using 23 DCM, 21 HCM, 11 HHD, and 13 non-hypertrophic hearts, the incidence of the in situ markers showed significant correlations with the left ventricular mass index and myocyte size, but not with cardiac function and dilatation. Positivity of myocytes for in situ markers for DNA breaks, such as TUNEL and in situ ligation assays, may be an epiphenomenon accompanying cardiac hypertrophy, but not myocyte death in pathological human hearts. PMID- 12533837 TI - Mutations of mtDNA in renal cell tumours arising in end-stage renal disease. AB - Toxic effects in the uraemic state or during maintenance dialysis have been suggested to be responsible for DNA damage and tumour development in end-stage renal disease (ESRD). This study therefore analysed the mitochondrial DNA alterations in six kidneys with ESRD and in nine renal cell tumours arising in these kidneys. Sequencing the entire 16 569 bp mitochondrial genome disclosed 94 sequence variations in normal and corresponding tumour tissues. Thirty-eight polymorphisms occurred in the D-loop region, 40 in the polypeptide coding regions, 12 in the rRNAs, and four in the tRNAs. Nine somatic nucleotide changes were found in seven of the nine tumours analysed; four of them were G to A transitions. Two of the G to A changes occurred in the D-loop region, one in the MTTA gene, and one in the MTND2 gene. An A to G substitution was seen in the control region at the mtTF1 binding site. A T to C transition also occurred also in the D-loop region. A T insertion was seen in MTRNR2 (16S rRNA). One C insertion in MTND4 and one A deletion in the polyA tract of the MTND5 gene resulted in frameshift mutations in two tumours. This study reveals a high mutational rate of the mitochondrial DNA in tumours, which may correspond to the increased level of reactive oxidative species in renal parenchymal cells in ESRD. PMID- 12533838 TI - Papillary carcinoma of the thyroid: evidence for a role for hepatocyte growth factor (HGF) in promoting tumour angiogenesis. AB - The pattern of vascularization of papillary carcinoma was investigated in tumour sections from 31 cases and in primary cultures from 12 cases. Tumour sections were immunostained for von Willebrand Factor (vWF) to visualize blood vessels; for endothelial-specific nitric-oxide-synthase (EC-NOS), as a marker of endothelial cell activation; and for Ki-67 to evaluate endothelial cell proliferation. It was found that endothelial cells lining venous vessels located in peritumoural fibrous tissue were intensely EC-NOS-positive and occasionally Ki 67-positive. Capillary vessels of tumour papillae were not stained for Ki-67 and were weakly EC-NOS-positive. Primary cultures of papillary carcinoma cells were used as a potential source of factors active on endothelial cells. It was found that thyroid tumour cells contain RNAs for angiopoietin, vascular endothelial growth factor (VEGF), and VEGF-C; moreover, they release large amounts of VEGF into culture supernatants and exert chemotactic activity in vitro for the endothelial cell line SIEC. The ability of papillary carcinoma cells to release angiogenic factors could be stimulated in vitro. Hepatocyte growth factor (HGF; 25 ng/ml) induced a 1.2- to 5-fold increase in the amount of VEGF released by tumour cells and a 1.2- to 4.2-fold increase in the amount of chemotactic activity present in culture supernatants. Met protein, the high affinity HGF receptor, is overexpressed in a large proportion of cases of papillary carcinoma. These findings are consistent with the possibility that HGF-Met protein interaction is one of the molecular mechanisms promoting the vascularization of papillary carcinoma of the thyroid. PMID- 12533839 TI - Nuclear expression of Y-box-binding protein-1 correlates with P-glycoprotein and topoisomerase II alpha expression, and with poor prognosis in synovial sarcoma. AB - Nuclear expression of the Y-box-binding protein (YB-1) has been reported to correlate with the expression of P-glycoprotein in breast cancer and osteosarcoma. Overexpression of the ATP-binding cassette (ABC) superfamily, such as P-glycoprotein/multi-drug resistance (MDR) 1 and MDR-associated protein (MRP) 1, 2 and 3, has been reported in various malignant neoplasms. Fifty-four surgically resected synovial sarcomas were examined immunohistochemically for nuclear expression of YB-1 and intrinsic expression of P-glycoprotein, MRP1, MRP2, and topoisomerase II alpha, and the findings were compared with clinicopathological parameters, proliferative activities as evaluated by MIB-1 labelling index (LI), and the patients' prognoses. In addition, MDR1, MRP1, MRP2, and MRP3 mRNA levels were assessed using a quantitative reverse transcriptase polymerase chain reaction (RT-PCR) method in 22 concordant frozen specimens from these cases and the findings were compared with six control skeletal muscle tissues. Independent prognostic factors were investigated using the Cox proportional hazards regression model. Nuclear expression of YB-1 protein correlated with P-glycoprotein expression (p = 0.0126). Moreover, cases with nuclear expression of YB-1 correlated with poor survival (p = 0.0495) and showed a high topoisomerase II alpha labelling index (topo II alpha LI) (p = 0.0056) and a high MIB-1 LI (p = 0.01). Multivariate Cox analysis showed that only the nuclear expression of YB-1 (p = 0.0136) and high American Joint Committee on Cancer (AJCC) stage (ie stage III or IV) (p < 0.0001) were independent factors for poor prognosis, while the expression of the YB-1 responsive gene products examined was not. These results indicate that the nuclear expression of YB-1 protein is associated with P-glycoprotein expression and proliferative activity as shown by the topo II alpha LI and the MIB-1 LI, and that expression of this protein is an important independent prognostic factor in synovial sarcoma. PMID- 12533841 TI - Re: Tomlinson et al. Does MSI-low exist. J Pathol 2002; 197: 6-13. PMID- 12533840 TI - NEDD8 protein is involved in ubiquitinated inclusion bodies. AB - Proteolysis by the ubiquitin-proteasome system is considered to play a pathological role in several degenerative diseases that involve ubiquitinated inclusion bodies. In recent years, several ubiquitin-like proteins have been isolated, but it is uncertain whether their roles are associated with protein degradation through the ubiquitin-proteasome system. NEDD8 (neural precursor cell expressed and developmentally down-regulated gene), which consists of 81 amino acid residues, possesses the highest sequence similarity to ubiquitin. Recent studies have indicated that NEDD8 is covalently ligated to cullin family proteins, which are components of certain ubiquitin E3 ligases, by a pathway analogous to that of ubiquitin. Thus, by focusing on the structural and functional association between NEDD8 and ubiquitin, it would be of interest to know whether the NEDD8 system is involved in pathological disorders of the ubiquitin-proteasome system. This study has examined the immunohistochemical distribution of NEDD8 protein by using a highly purified antibody in normal tissues and in tissues known to contain ubiquitinated inclusions. NEDD8 protein expression was widely observed in most types of tissues. Furthermore, accumulation of the NEDD8 protein was commonly observed in ubiquitinated inclusion bodies, including Lewy bodies in Parkinson's disease, Mallory bodies in alcoholic liver disease, and Rosenthal fibres in astrocytoma. Two of ten cases of neurofibrillary tangles and senile plaques from patients with Alzheimer's disease showed intense staining for NEDD8 as well as for ubiquitin. These findings suggest the possibility that the NEDD8 system is involved in the metabolism of these inclusion bodies via the ubiquitin-proteasome system. PMID- 12533843 TI - What to expect at your eye exam. PMID- 12533844 TI - Smart shopping for a healthy heart. PMID- 12533845 TI - Grandparenting. How to help. PMID- 12533846 TI - Quackery. Spotting it and avoiding it. PMID- 12533847 TI - Cooking from the diabetes food pyramid. Meat and other proteins. PMID- 12533848 TI - Urinary tract infections. Treatment and prevention. PMID- 12533849 TI - For parents. Planning a vegetarian diet. PMID- 12533850 TI - What your doctor is reading. Caring for kidneys. PMID- 12533851 TI - Today's exercise and fitness trends. PMID- 12533852 TI - Diabetes quiz. How much do you know about treating impotence? PMID- 12533853 TI - Sweeteners. Herbal and otherwise. PMID- 12533855 TI - Supermarket smarts. Fish. PMID- 12533854 TI - Women & diabetes. Menopause. The latest on hormone replacement therapy. PMID- 12533856 TI - Dealing with surgery. PMID- 12533857 TI - [A study on the relationship between postmortem interval and the changes of DNA content in the kidney cellule of rat]. AB - OBJECTIVE: To study changes of DNA content in the kidney cellule of rats and relationship with the postmortem interval. METHODS: This experiment chose seven parameter of cell nuclear, including the area and integral optical density, determined the changes of DNA content in the kidney cellule of 15 rats at different intervals between 0 and 48 h postmortem with auto-TV-image system. RESULTS: The degradation rate of DNA in nuclear has a certainty relationship to early PMI(in 48 h) of rat, and get binomial regress equation. CONCLUSION: Determining the quantity of DNA in nuclear should be an objective and exact way to estimate the PMI. PMID- 12533858 TI - [A study on pathological changes of closed diffuse brain injury in rats with H.E., esterification-silver stain and albumin immunohistochemical stain]. AB - OBJECTIVE: To observed the pathological changes of closed diffuse brain injury in the rats died immediately and 15 min to 5 days after the injury. METHODS: H.E. staind and esterification-silver stain were applied to investigate the closed diffuse brain injury. RESULTS: In rats died immediately after the concussive injury, a number of shrunken neurons(type I change), distended neurons(type II change) and wave-like nerve fibers were identified in the brain tissue, especially in brain-stem. At 2 h and 8 h after injury, brain edema and axonal swelling appeared clearly in the cortex and white matter, especially in brain stem. At post-traumatic 8 h and 24 h, the axonal retraction balls began to appear. The amount of neurons undergoing type I and II changes and constraction balls increased along with the survivor time. After 4 days and 5 days, brain edema alleviated, but the retraction balls and axonal swelling still existed. With Esterifica-tion silver stain, the above changes of neurons and nerve fibers were more obvious. With ABC stain, the distribution of albumin(Al) was extended from the perio-vascular area to diffuse distribution. Al positive staining were more obvious in injuried neurons and nerve fibers. CONCLUSION: The distribution of the concussive damage in the brain are coup, contra-coup and centripental. PMID- 12533859 TI - [Detection of coxsackie virus B3 gene in myocardium and spleen samples in myocarditis with sudden death]. AB - OBJECTIVE: To detect the Coxsackie virus B3(CVB3) gene in myocardium and spleen tissues in viral myocarditis(VMC) with sudden death and to explore the diagnostic method for VMC by means of seeking pathogene. METHODS: By in situ RT-PCR, the detection of CVB3 gene in myocardium and spleen sections were performed in sudden death group caused by VMC and non-cardiac death group. RESULTS: In VMC group, CVB3 gene-positive signals were seen in myocardium sections(3 out of total 8 cases, No. 1, 4, 7 cases) and spleen sections(4 out of total 8 cases, No. 2, 4, 6, 7 cases). In non-cardiac death group, no positive signals were detected in both myocardium and spleen tissues. CONCLUSION: Positive detection of CVB3 gene in both myocardium and spleen maybe an important character of VMC and can improve the detecting pathogene in diagnosing VMC. PMID- 12533860 TI - [Study on the relationship between postmortem interval and the change of absorbance in vitreous humor of rabbit after death]. AB - OBJECTIVE: To seek a exact method of estimating Postmortem interval (PMI). METHODS: This study was preformed to investigate the relationship between postmortem interval and absorbance in vitreous humor of rabbit after death. The absorbance in vitreous humor of 48 rabbits after death were investigated with Model 754 spectrophotometer in apt wavelength (420 nm). RESULTS: There exists positive linear regression association between postmortem interval (Y) and absorbance in vitreous humor (X) (r = 0.98327, P < 0.05), during rabbits after death 0 to 72 hours. The formula of linear regression is Y = 453.30 X + 0.75 (Y = postmortem interval = PMI, X = absorbance in vitreous humor). CONCLUSION: The absorbance in vitreous humor can be as reference indicator to estimate PMI within hour 72. PMID- 12533861 TI - [Study of the mechanism of cultured neuron injury mediated by nitric oxide during hypoxia and oxidative stress]. AB - OBJECTIVE: To study the mechanisms of cultured neurons injury mediated by nitric oxide and free oxygen radical during hypoxia and oxidative stress. METHODS: The cultured newborn rat neurons were treated with hypoxia, H2O2 and pretreated superoxide dismutase (SOD) respectively. We examined the content of NO, malonaldehyde (MDA), lactate dehydrogenase (LDH) and SOD in cultured supernatant. RESULTS: Comparing with that of control group, the content of NO, LDH, MDA increased and the content of SOD decreased in hypoxia group and H2O2 group. The content between NO and SOD showed the negative correlation. Administration of 200 U/ml SOD before oxidative stress could efficiently decrease the release of NO, LDH and MDA in neurons. The content of NO, LDH and MDA manifested in positive correlation in each group. CONCLUSION: Hypoxia and oxidative stress increased NO production which strengthen neurons injury induced by free radical. SOD played an important role in elimination of free oxygen radicals and protecting neurons from injury by NO. PMID- 12533862 TI - [Genetic studies of 13 loci in Guangdong Han population by two multiplex PCR systems and its forensic application]. AB - OBJECTIVE: The genetic studies of 13 short tandem repeats(STRs) loci in two multiplex amplification systems were carried out on Chinese Han population in Guangdong. METHODS: DNA samples from 328 unrelated individuals were screened. The 13 loci were D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820, D16S539, TH01, TPOX and CSF1PO. The PCR products were analyzed and genotyped by ABI 377-96 Sequencer. RESULTS: The combined power of discrimination (DP) was 0.999999999999993 and the combined paternity of exclusion(PE) was 99.999%. These 13 loci met the Hardy-Weinberg expectations. CONCLUSION: The two multiplex amplification systems were very useful in forensic case investigation. PMID- 12533863 TI - [Study on application of CODIS loci to excluding paternity]. AB - OBJECTIVE: The study was carried out on application of CODIS loci (FGA, vWA, CSF1PO, TH01, TPOX, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51 and D21S11) in 100 cases of excluding paternity. METHODS: The PCR amplified products of Profiler Plus and Cofiler amplification kit were injected into a capillary on the ABI PRISM 310 Genetic Analyzer. GeneScan software analyzed the collected data, which can then be imported into Genotyper software for genotyping of alleles. RESULTS: In the group of Mother-Child-Alleged Father, more than 3 STR loci incompatibilities between alleged father and child were found in all observed cases, the mean incompatibility was 6.63; In the group of Alleged Father Child, 94.0% of all cases observed was found more than 3 STR loci incompatibilities, the mean incompatibility was 5.01. CONCLUSION: The results showed that CODIS loci had good application to excluding paternity, and the choice of hyperpolymorphic markers in investigation, which were valued by DP, H and PE, had direct relation with the augmentation of incompatibilities in excluding paternity. PMID- 12533864 TI - [Chinese Hans population study of fluorescent labelled 8 STR multiplex system]. AB - OBJECTIVE: Chinese Hans population study of 8 STR loci was carried out in 350 unrelated individuals. METHODS: The 8 STR loci(vWA, TH01, TPOX, CSF1PO, D5S818, D13S317, D7S820, D16S539) and a sexual distinction locus Amelogenin, were amplified simultaneously in one PCR reaction tube, the products were separated by electrophoresis with PE-377 DNA sequencer, and analyzed by Genescan analysis software, then the accurate PCR fragment length and designation of genotypes were finished automatically by the sequencer. RESULTS: 63 alleles and 2 sex determination alleles were found in Hans population altogether; the minimum allele frequency in Chinese Hans population at these loci was 0.17%, the maximum was 50.17%; the fragments were 119-327 base pairs long; the TDP of this system reached 99.99999998%; this system inherited steadily, and was accord with Mandel inheritance rule, no gene mutation was found, all loci met Hardy-Weinberg equilibrium, but the frequencies of Hans population differed from those of African-Americans, Caucasion-Americans and Hispanic-Americans. CONCLUSION: This method is good for identification of little amount and degraded DNA samples from blood stain, semen stain mixed with vaginal excretion, hair root, cigarette root, muscle tissue and ancient bone. It can also make a foundation of establishing the National DNA Database. PMID- 12533865 TI - [Recent progress of study on single nucleotide polymorphisms]. AB - Single nucleotide polymorphisms (SNPs) is a new genetic marker system following RFLP and microsatellite polymorphism. It has been shown to have the characteristics of high-density, inheritance stability and facilitation in analysis automation. SNPs can be detected by electrophoresis, endonuclease cleavage, PCR and sequencing, and can be used extensively in gene mapping, disease-correlativity analysis, population genetics and drug research. PMID- 12533866 TI - [In vivo distribution and expression after skin injury of fibronectin splicing variants]. AB - OBJECTIVE: In order to develop new criteria and to select positive controls for forensic medicine, experiments on in vivo distribution and expression after skin wound of fibronectin splicing variants, EIIIA and EIIIB were studied. METHODS: Rat tissues were homogenized to extract total RNA which was then amplified by RT PCR technique. RESULTS: (1) EIIIA+ and EIIIB+ of normal skin were not expressed, whereas EIIIA+ after 18 hours' injury was expressed abundantly. No obvious charge of EIIIB+ was detected. The differences between results of using two methods of injury were not significant. (2) The distribution of fibronectin variants in different adult organs were in the following order: brain > heart > liver. CONCLUSIONS: EIIIA+ may be used as a sensitive criterion for research of skin injury. PMID- 12533867 TI - [Fos expression in rat skin after different incised wound healing interval]. AB - OBJECTIVE: To investigate the expression of c-fos in rats' skin during wound healing. METHODS: Immunohistochemistry was conducted on paraffin section from incised wounding model of rat skin. RESULTS: Fos protein improved from the time of 10 min after wounding in the wound edge, then it reached peak at 3 h. 24 h after injury, the quantity of Fos expression had no difference with that of normal skin. CONCLUSION: Fos is sensitive after wound, but should be used with other criteria in wounding interval estimation as it's unstediness. PMID- 12533869 TI - [Experimental study of restiffening of the rigor mortis]. AB - OBJECTIVE: To observe changes of the length of sarcomere of rat when restiffening. METHODS: We measured the length of sarcomere of quadriceps in 40 rats in different condition by scanning electron microscope. RESULTS: The length of sarcomere of rigor mortis without destroy is obviously shorter than that of restiffening. CONCLUSION: The length of sarcomere is negatively correlative to the intensity of rigor mortis. Measuring the length of sarcomere can determine the intensity of rigor mortis and provide evidence for estimation of time since death. PMID- 12533868 TI - [A preliminary study on the changes of expression of PDGF-beta, PDGFR-beta, TGF beta 1, TGFR, bFGF and its relationship with the wound age in wound healing]. AB - OBJECTIVE: To explore the relationship between the expression change of cytokines and the wound age during the healing process of rats skin wound. METHODS: Immunohistochemical and image-analysis methods were performed on vital skin wounds(after incision 0.5-168 h am) and postmortem damage(after incision 0.5-6 h pm). RESULTS: The expression of the cytokines PDGF-beta, PDGFR-beta, TGF-beta 1, and bFGF in the epithelial cells was already enhanced since 0.5 h am after damage and their strongest expression reaction was seen at 24-96 h am. In addition, the expression of PDGF-beta, PDGFR-beta, TGF-beta 1 and bFGF was also found in the macrophages and the fibroblasts of the granulation tissue, and the expression changes in the postmortem damage group showed that the skin tissue within 0.5-3 h after incision showed immunohistochemical changes but weakly expression and 3 h thereafter no any change was found. CONCLUSION: The expression characteristics of the above mentioned cytokines in wound repair should be related to the wound age and it reminds therefore that they may be used as immunohistochemical criteria for accurate determining the wound age. PMID- 12533870 TI - [The preliminary study on change of serum enzyme level in crush syndrome rat]. AB - OBJECTIVE: To study the changes of serum creatinine kinase(CK) and its cardiac specific isoenzyme compound(CK-MB) levels in crush injury rats. METHODS: Crush injury was produced in SD rats, the serum levels of CK and CK-MB were studied by automated biochemical analyzer. RESULTS: The levels of plasma CK and CK-MB were much higher in crush injury rats than those of the control group. CONCLUSIONS: Cardiomyocyte injury may be induced in the early stage of crush injury rats. PMID- 12533871 TI - [Diagnosis of non-typical myocarditis by applying immunohistochemical method of CD68]. AB - OBJECTIVE: To make a distinction between myocarditis and the reaction to some pathological state of myocardium. METHODS: Myocardium of 26 cases with sudden cardiac death were stained and LM light microscopies with immunohistochemical method 10 cases with normal myocardium were contrasted. RESULTS: A great deal of stained positive monocyte of immunohistochemistry emerged in the parasetions of myocarditis patients with various farms and stacking(> 15). CONCLUSION: The stain of immunohistochemistry can be used as one of the indications for diagnosing non typical myocarditis. PMID- 12533872 TI - [Effects of perming, combing and stretching on hair keratins]. AB - OBJECTIVE: Research of the hair damage due to perming, combing and stretching can be of important value for forensic hair individual identification. METHODS: The normal human hairs were treated with perming combing and stretching, and the keratins of the damage hair were analysed by using SDS-PAGGE and laser densimeter. RESULTS: Perming, combing and stretching brought about hair damage; The keratins of the damage hair were obviously reduced at the rang of molecular weight of 67,000-43,000 dalton. CONCLUSION: The loss of the damage hair keratins were increased with the degree of the hair damage. PMID- 12533873 TI - [The changes of tryptase activity in anaphylactic shock guinea-pigs]. AB - OBJECTIVE: To investigate the changes of the activity of tryptase of sera, lungs and bronchial tubes in the guinea-pigs which suffered from hetero-serum anaphylactic shock. METHODS: Sera and tissues were collected from anaphylactic shock guinea-pigs, and the enzyme activity was tested colormetrically using special substrate, BAPNA. RESULTS: The activity of tryptase of sera, lungs and bronchial tubes increased significantly in Anaphylactic guinea-pigs compared with control group. CONCLUSION: The changes of tryptase activity are helpful to diagnose anaphylactic shock. PMID- 12533874 TI - [Study of distribution and metabolism of methamphetamine in hair of guinea pig]. AB - OBJECTIVE: To study the distribution and metabolism of methamphetamine in the hair of guinea pig. METHODS: Determination of methamphetamine and its metabolite amphetamine in hair was performed by GC/MS and GC/NPD. Concentration-time course of methamphetamine and amphetamine in hair of guinea were recorded. Relationship between hair color, administrated dose and drug concentration in hair were also discussed. RESULTS: The concentration of amphetamine is higher than the concentration of methamphetamine in the hair of guinea administrated a single dose or seven doses of methamphetamine. The concentration of methamphetamine and amphetamine were significantly related with administration dose and the incorporation rate into white and brown hair is much poorer than that of black hair. CONCLUSION: Administration methods, dose and the color of hair affect the concentration of methamphetamine and amphetamine. PMID- 12533875 TI - [The application of vascular endothelial growth factor in forensic science]. AB - Vascular Endothelial Growth Factor (VEGF) is a potent angiogenic factor with a key role in several pathological processes, including wound repair as well as a effective vascular permeability factor. This article review the present study of VEGF in molecular biology, the connection with repair and expression regulation and so on. PMID- 12533876 TI - [The research progress of the association of mitochondrial DNA mutation with cardiomyopathy]. AB - There are some human diseases associated with mitochondrial DNA genome defect. Now many studies think that: oxygen radical resulting from oxidative phosphorylation(OXPHOS) disorder caused by myocardium ischemia and the increased OXPHOS induction damage mitochondrial DNA. Chronic damage accumulations lead to mitochondrial DNA deletion or point mutation in the end which show mitochondrial DNA 5.0 kb or 7.4 kb deletion and point mutation at position C15452A in the cytochrome b gene; the conservative sequence mutation of tRNA gene such as A4300G, C4320T point mutation in the tRNA Ilegene, A3243G point mutation in the tRNA leu gene etc result in defective contractile proteins whose persistent and inefficient contraction may increase the myocardium's metabolic demands for ATP and leads to cardiac hypertrophy. In this article, we review the study on the association of mitochondrial DNA mutation with ischemic cardiomyopathy and hypertrophic cardiomyopathy. PMID- 12533877 TI - [The alternative splicing & expression of fibronectin IIIcs segment and its relationship with wound healing]. AB - Fibronectin is an important large adhesive glycoprotein of the extracellular matrix, which is alternatively spliced in three regions, designated EIIIA, EIIIB and IIIcs respectively. IIIcs contains two binding domains for a variety of cell surface and extracellular ligands. Through this multiplicity of adhesive activities, IIIcs can fulfill key roles in a broad spectrum of physiological processes, such as cell spreading and migration, differentiation and embryogenesis, wound healing, malignant transformation and metastasis, etc. Here, we will discuss the structure, biological property, and function of IIIcs splicing variants and its forensic applications. PMID- 12533878 TI - [The single nucleotide polymorphisms and its application to forensic medicine]. AB - Single-nucleotide polymorphisms (SNPs) are the most abundant forms of human genetic variation. These variable sites are present at high density in the genome, making them powerful tool for the diagnosis of genetic and genetic related diseases, population genetics research and drug development. They are also found widespread application to the forensic medicine. This report mainly describe the SNPs characteristics and its potential applications to the forensic medicine including the possibility, the problems and high-throughput automation detection methods. PMID- 12533879 TI - [Imaging diagnosis and forensic appraisal of orbital fracture]. AB - Misdiagnosis and missed diagnosis are common in forensic appraisal of orbital fracture. Now imaging technology is very important for studying the forensic features of orbital fracture and evaluating the degree of injury. This article reviews the classification, pathogenesis and imaging diagnosis of orbital fracture. It may do some help to forensic appraisal of orbital fracture. PMID- 12533880 TI - [Study on the expression of fibronectin after cerebral contusion in rats for timing of injuries]. AB - An experimental model of reproducible focal cerebral contusions in rats was made by a free-drop impacting right hemisphere. The expression of fibronectin and its mRNA after cerebral contusion were detected respectively by immunohistochemical staining and in situ hybridization. Results indicated that the expression of fibronectin and its mRNA increased after injury, and there existed a relationship between increased fibronectin and its mRNA and different intervals after brain injury. It is inferred that the expression of fibronectin and its mRNA can be used for timing of brain injuries and distinguishing antemortem and postmortem brain contusions. PMID- 12533882 TI - [Study on the specificity of complement C5 for the postmortem diagnosis of myocardial infarction]. AB - In order to explore the specificity of complement C5 in the postmortem diagnosis of myocardial infarction, changes of C5 staining in normal, infarcted and other non-infarcted myocardia with direct or indirect myocardial injuries (myocarditis, mechanical asphyxia, electrocution, hemorrhagic shock, cardiac contusion and organophosphate poisoning) were studied with immunohistochemistry and image analysis. The results showed that positive C5 staining could be observed in groups of myocardial infarction and myocarditis, but not in groups of mechanical asphyxia, electrocution, hemorrhagic shock, cardiac contusion, and organophosphate poisoning. It is indicated that positive reaction of C5 could only be affected by myocarditis, which means that it was more specific for the diagnosis of myocardial infarction. PMID- 12533881 TI - [Histopathological observation of experimental slight viral myocarditis]. AB - To study the diagnostic method of slight viral myocarditis in the field of forensic pathology, slight viral myocarditis model was induced in Balb/c murine by coxsackie virus B3. Organs of hearts, livers, spleens, lungs and kidneys were examined through routine pathological methods. Pathological changes at different levels of these organs were observed. The results indicated that viral myocarditis was a kind of disease with multiple organ alterations and that the pathological observation and comprehensive analysis of multiple organs was one of the useful methods for diagnosing slight viral myocarditis. PMID- 12533883 TI - [Histochemical observation of changes of neurons and axons in injured brainstem]. AB - Brainstem of rats were stabbed with a needle and pathological changes of neurons and axons in brainstem were observed at different time after injury with Nissl's body staining, silver staining and modified trichrome staining. It was found that, by silver staining, the axons showed irregular swelling and disconnection at 1-3 h, marked swelling of the severe end at 6 h, retraction ball at 15 h and remarkable retraction ball at 24 h. By modified trichrome staining, the space between myelin sheaths and axons was widened at 3-6 h, and tortuous myelin sheaths adhered incompletely on axons, or even peeled off at 15 h to 24 h. Perinuclear lysis of Nissl's bodies at 24 h after injury could be seen by Nissl body staining. The results indicated that, the pathological changes in injured brainstem could be observed with histochemical staining, which might be used for timing brainstem injuries. PMID- 12533884 TI - [Forensic application of reverse dot blot with biotin incorporation]. AB - A rapid and accurate reverse dot blot technique was successfully established. It could distinguish the 6 alleles of 0101, 0102, 0103, 0201, 0301 and 0401 of HLA DQA locus. Alleles and genotype frequencies for HLA-DQA locus were determined in 200 unrelated individuals of northern Han population. The discrimination power (DP) was 0.928, heterozygocity (H) 0.75. By direct biotin incorperation, 1 ng genome DNA could be detected successfully. This method can be used for paternity test and individual identification in forensic science practice. PMID- 12533885 TI - [A study of genetic polymorphism of the STR locus D2S441 in Chinese Han population in Chengdu]. AB - Using PCR and PAG, followed by silver staining, the tetrameric STR D2S441 locus was studied in 260 unrelated Chinese individuals living in Chengdu. 9 alleles and 26 genotypes were observed. The range of fragment size was 131 bp to 155 bp. The genotype distribution of D2S441 locus in Han population was in accordance with Hardy-Weinberg equilibrium. Family survey confirmed Mendelian inheritance of alleles. The discriminating power (Dp), observed heterozygosity (H), polymorphism information content (PIC) and power of exclusion (PE) were 0.9084, 0.7885, 0.7390 and 0.5778 respectively. The results demonstrated that this locus was highly polymorphic and could be used for forensic identification and paternity testing. PMID- 12533886 TI - [Cantonese population data of the tetranucleotide repeat locus D7S809 and its application in paternity test]. AB - The tetranucleotide repeat locus D7S809 was investigated in Cantonese population by polymerase chain reaction (PCR) amplification, subsequent polyacryramide gel electrophoresis and silver staining. 14 alleles and 50 genotypes were detected in 190 sample individuals. All alleles differed in size by 4 bp repeat. No interalleles were found. The heterozygosity, power of discrimination and chance of non-paternal exclusion were 0.8613, 0.9645 and 0.7184 respectively. No significant deviations from Hardy-Weinberg equilibrium were found. The D7S809 was a highly polymorphic, stable and accurate type locus and had been successfully applied to 100 paternity test cases. PMID- 12533887 TI - [Prospective study of lethal blood concentration of organophosphorous in humans]. AB - Total cases of organophosphorous (dichlorvos, methamidophos, dimethoate) poisoning outpatients from six hospitals during four years were collected consecutively for lethal blood concentration study. Blood samples were detected with gas chromatography. The probabilities of death, coma were analyzed with Bliss method and their linear regressive equations of probit were obtained respectively, their 50% lethal concentrations (LC50) and 50% coma concentrations(CC50) were calculated by the formulas above. As the death rate was influenced by therapy, its natural death probability has been discussed and estimated their natural LC50 were between the LC50 and CC50 themselves. Combined LC50 and CC50, their natural LC50 were calculated. PMID- 12533888 TI - [The forensic determination of post-traumatic epilepsy in 21 patients]. AB - To evaluate the forensic determination of post cerebral traumatic epilepsy. METHODS: In 21 patients, traumatic history and previous history were analysied combined with the demonstrations of electroencephalogram(EEG), X-ray, CT and MRI. RESULTS: Post-traumatic epilepsy, manily in late stage, usually occurred following serious cerebral trauma. The type of traumatic epilepsy was determined by the traumatic location and extent. Abnormal epileptic wave in scalp EEG and 24 h dynamic EEG and medical image examinations were helpful for qualitative analysis. CONCLUSION: The forensic determination of post traumatic epilepsy must be on the basis of traumatic and previous history combined with EEG, CT and MRI analysis. PMID- 12533889 TI - [Analysis on characters of 220 cases of tympani membrane perforated]. AB - 220 cases of tympani membrane perforated were summarized retrospectively. The tympani membrane perforated of injury was different from otitis media's because they had marked or extreme marked difference in sex, age, ear distinction, shape, position, size, degree of hearing damaged, time of healing, cause of perforating and adhesive substance. This difference was relative to mechanism of tympani membrane perforated. The distinction gist of two sorts of tympani membrane perforated was expounded through comparison and analysis. It would be of great value to clinical forensic medical examination. PMID- 12533890 TI - [Study on the relationship between PMI and the concentration of zincum and nickel in the vitreous humor of rabbit after death]. AB - OBJECTIVE: To seek a exact method of estimating postmortem interval (PMI). METHODS: This study detected the concentration of zincum(Zn) and nickel(Ni) in vitreous humor of rabbit at hour 96 after death and explored the relationship between their concentration and PMI using a method ICP-MS. RESULTS: The concentration of Zn and Ni in vitreous humor of rabbit at hour 24 after death were related to PMI significantly; The formulae of the relationship between PMI and Zn concentrations is y = 0.1404x2 - 1.3351x + 3.8298 (within 24 h; R2 = 0.9202). The formula of the relationship between PMI and Ni concentrations is y = 0.0043x2 - 0.0596x + 0.2665(within 24 h; R2 = 0.9103). CONCLUSION: The concentration of Zn and Ni in vitreous humor of rabbit may be a reference indicator to estimate early PMI. PMID- 12533891 TI - [A study of the relation between interval of the injury and the reaction of neurons, glial cells after experimental brain contusion]. AB - OBJECTIVE: To observe the change of c-fos protein(Fos) and nerve growth factor receptor (NGFR) staining in the brain of rat after experimental brain contusion. METHODS: Immunohistochemistry of c-fos and NGFR were applied to investigate the brain contusion. RESULTS: (1) The expression of Fos protein could be observed at 0.5 h after injury and then increased with the prolonging of time. By 3 h after injury, the positive staining cells could be detected massively not only in and round the wound site but also in other areas of the whole ipsilateral cortex. The stains decreased 6-12 h later and could hardly be detected 1 d after the brain contusion. The control-experiment is negative. (2) NGFR positive staining cells could be found round the wound area 1 d postlesion. At 3 d following injury, a peak of massive positively stained cells appeared both in number and in intensity, showing significant differences compare with that of 1 d after damage (P < 0.01). 5 d later the positive express declined slowly. The express in the control-rat is negative. CONCLUSION: There is a rule that the expression of Fos and NGFR positive staining changes with time going after brain contusion, which will be of great value in estimation of brain injury time. Detection of Fos can be used for time deduction in earlier period after injury, while NGFR in later period. They are also very important for distinguishing between antemortem or postmortem injury. PMID- 12533892 TI - [Study on expression of the glycoprotein in myocardial cell membrane of the rat's early myocardial ischemic]. AB - OBJECTIVE: To explore expression of the glycoprotein in early myocardial ischemic. METHODS: The glycoprotein changes occurred at the early acute cardiac ischemic area induced experimentally by ligation of left coronary artery of 32 SD rats. 6 lectins were measured by means of immunohistochemical methods. RESULTS: Positive staining of PNA could be observed in ischemic area at 5 min after ischemia, and the positive area increased with the prolongation of ischemic period. It became the strongest for 2 h and then decreased. CONCLUSION: This experiment proved that myocardial cell membrane in ischemia expressed D galactose. This may be of some value in forensic medicine practice. PMID- 12533893 TI - [Histological study of species identification using bones]. AB - OBJECTIVE: To explore the species identification methods of bone remain. METHODS: The compare between human bone and common animals bones was made with microscopic comparison methods. The sample was consisted of 77 slides including human, cattle, house, pig, goat, sheep, monkey, cat, rabbit, chicken, duck, pigeon, tortoise, frog, fish. RESULTS: The differences of the histological bone species were as follow: the thick of the outer and the inner circumferential lamellae, and the shape of interstitial lamellae, and the size of Haversian system, and the number of the circumferential lamellae, and the number of the lacunae of inner Haversian system. CONCLUSION: The histological characters of the bones can be used for species identification. PMID- 12533894 TI - [Ultrastructure changes of electrical injury in rats]. AB - OBJECTIVE: To observe ultrastructure changes of electrical injury in rats. METHODS: An experimental model of rats suffered from the low voltage were designed. Ultrastructure changes of electrical injured tissues were observed under transmission electron microscope. RESULTS: (1) Plasma of epithelium was concreted in the affected areas and inner membrane system was broken. (2) Hypercontraction bands were observed in skeleton muscles. (3) There was dissolved necrosis and hypercontraction bands in the myocardium. (4) Vacuoles were found in plasma of endothelium of blood vessels on electrical current path, and myelin sheath of nerve fiber were loosed. CONCLUSION: The above mentioned ultrastructure changes could be used as assistant diagnostic index of electrocution. The mechanism of the changes were discussed. PMID- 12533895 TI - [Research of ultra-structural pathological changes of nervous, endocrine and immune system in heroin addicts]. AB - OBJECTIVE: To investigate ultrastructural pathological changes of Heroin-Addicts. METHODS: Heroin-Addicts' central nervous system, endocrine system, immune system and reproductive system in 4 cases are observed by using transmission electron microscope(TEM). RESULTS: The changes of central nervous system are mitochondrion swelling, crista fragmentation and disappear. Endoplasmic reticulum dilation, nervous fibres and cell organelles reduction; mitochondrion swelling, Partial crista fragmentation and endoplasmic reticulum dilation are also found in endocrine system; Lymphocytes reduction, cytoplasm ingredient reduction and dead lymphocytes increase in immune system; in reproductive system, spermatogenic cells and cell organelles are reduced in the male and follicle disappeared in the female. CONCLUSION: Ultra-structural pathological changes of heroin-addicts are presented acute, chronic oxygen deficiency degeneration and necrosis. PMID- 12533896 TI - [Study on the construction of standard D1S549 allelic ladder via molecular cloning and its genetic polymorphism in Chinese three populations]. AB - OBJECTIVE: To resolve the problem of the accuracy and standardization of STR-PCR typing in forensic practice, we have designed a new method to produce standard D1S549 allelic ladder. METHODS: Eight different PCR amplified D1S549 allelic fragments were isolated from the gel, eluted into the distilled water and re amplified by PCR. The purified allelic fragments were then blunt-end subcloned individually into the pUC plasmid vectors and transfected into competent E. coli DH5 alpha cells. RESULTS: The sequencing results confirmed that the size and the construe of the inserts were correct. The recombinant plasmids DNA with 8 inserts were then used as templates for re-amplification to generate D1S549 standard ladder, with which the genetic polymorphisms of D1S549 locus in Chinese Han population in chengdu, Hui population in Gansu and Wei population in Xinjiang were studied. CONCLUSION: The results showed that the standard ladder made via this method is excellent, and D1S549 locus is robust for genetic research and forensic application. PMID- 12533897 TI - [Study on the forensic application of ABO genotyping by PCR-RFLP]. AB - OBJECTIVE: The study was carried out on the application of ABO genotyping by PCR RFLP to forensic purpose. METHODS: The DNA of samples were extracted by Chelex- 100 and phenol/chloroform, the PCR products were separated by Native Horizontal PAGE, and detected by silver staining. RESULTS: The PCR amplified fragment length of ABO loci is from 175 to 210 bp, the frequency distribution of six genotypes ranges from 0.0250 to 0.4300, heterozygosity(H) and discriminating power(Dp) is 0.5162 and 0.7111. CONCLUSION: This technique has a good application to the forensic samples such as blood, blood stain, semen stain, hair root, bone tissue and semen from vaginal swabs. PMID- 12533898 TI - [A study on polymorphism of salivary esterase in personal identification]. AB - OBJECTIVE: To research the value of polymorphism of salivary esterase(Set) in paternity and personal identification. METHODS: Phenotype and genotype of human salivary esterase were detected in 114 liquid saliva samples from the Chinese population by disc electrophoresis and fast blue RR staining assay. RESULTS: The frequency of Set type was F 22.81%, FS 50.88%, S2 6.31%. The estimated gene frequency of SetF was 0.4825 and SetS was 0.5175. The PE was 0.1875 and the DP was 0.6199. CONCLUSION: Polymorphism of salivary esterase (Set) was practical in paternity and personal identification. PMID- 12533899 TI - [Study on time limit of ACP and sperm positive-detected in mixed spot]. AB - OBJECTIVE: The study was carried out on time limit of ACP and sperm positive detected in mixed spot. METHODS: 600 vaginal swabs from living body after sexual intercourse were tested by routine and three-blindness methods. RESULTS: The longest time limit of ACP which was positively detected is 255 h, average time limit is 52 h, 41.3% of those samples was ACP-detected in positive degree (++). The longest time limit of sperm positively detected is 132 h, average time limit is 29 h. The number of sperms observed in 34.3% of those samples with positive result, is from 6 to 10/HP. CONCLUSION: This study is helpful to forensic identification of sex cases. PMID- 12533900 TI - [Percutaneous vertebroplasty--indications and procedural techniques]. PMID- 12533901 TI - [Efficacy of dynamic CT perfusion imaging in conjunction with three dimensional CT angiography for the evaluation of acute ischemic stroke]. AB - BACKGROUND AND PURPOSE: Through the use of a high-speed spiral CT scanner (GEMedical HiSpeedZX/i), CT/P/A technique, where conventional CT, CT perfusion imaging (CTP) and CT angiography (CTA) are consecutively performed, can now be performed with an imaging time of 90 seconds and a total contrast medium volume of 100 ml. A prospective clinical study was performed to ascertain the effectiveness of CT/P/A in diagnosing acute ischemic strokes. METHODS: 29 consecutive patients of Teraoka Memorial Hospital suspected of suffering from the occlusion or constriction of cerebral arteries and who underwent CT/P/A within 3 hours from the onset served as subjects. The sensitivity, specificity, or Odds ratio of CTP and CTA in detecting lesions that caused cerebral infarction was calculated. RESULTS: CTP detected a hypoperfusion area with a sensitivity, specificity, and Odds ratio of 80%, 64%, and 7.2. The sensitivity in lobar infarcts, white matter infarcts, basal ganglia infarcts, and brainstem infarcts was 100%, 100%, 100%, 0% (p = 0.0022). The sensitivity and Odds ratio of CT/P/A in cerebral infarcts differed according to the diameter of the infarcts. That with infarcts of 10 mm or more was 91%, 20. That with infarcts smaller than 10 mm was 50%, 2. CTA detected arterial lesions that caused cerebral ischemic attack with a sensitivity of 94% and specificity of 90%. The examination time for CT/P/A was 18 minutes, total radiation time being 90 seconds. CONCLUSIONS: Although CT/P/A was ineffective for the diagnosis of brainstem infarcts and lesions smaller than 10 mm, CT/P/A was useful in detecting moderate-sized hypoperfusion areas and arterial lesions three-dimensionally before an infarct is completed. PMID- 12533902 TI - [Surgical treatment of proximal middle cerebral artery (M1) aneurysms at the origin of the lenticulostriate artery]. AB - In contrast to aneurysms of the middle cerebral artery at the bifurcation, aneurysms at the origin of the lenticulostriate arteries (LSA) are uncommon. Six surgically treated patients (34 to 70 year-old; 3 men, 3 women) were reviewed. 5 patients presented with subarachnoid hemorrhage (H&H grade 2:3, 3:1, 4:1; Fisher type 2:1, 3:3, 4:1) and 2 patients had multiple aneurysms. All aneurysms arose from the postero-superior surface of the M1. Although neck clipping was achieved in every patient, re-application of the clip was necessary during surgery in 3 patients because the tip of the blade extended to the other perforators that ran parallel to the M1. Results were as follows: GR 3, MD1, SD 1, D1. Apart from a 70 year-old patient who died of vasospasm (H & H 4), fair results in two patients were accompanied by ischemic complications of the LSA. All 3 patients who required re-application of the clip during surgery showed a lacunar infarct of perforating arteries on post-operative CT. Special care of perforating arteries not only around the neck (the LSA) but also behind the aneurysm is essential for successful neck-clipping of aneurysms at this location. PMID- 12533903 TI - [Surgical stress in patients with asymptomatic unruptured intracranial aneurysms]. AB - BACKGROUND: Surgical stress elicits a characteristic response involving induction of synthesis and release of acute-phase proteins such as C-reactive protein (CRP). To objectively evaluate the surgical stress in patients with asymptomatic unruptured intracranial aneurysms, serum levels of CRP after direct clipping via the pterional approach were measured. METHODS: 42 patients who experienced an uneventful postoperative course, were evaluated according to age (less or more than 65 years old [adult or elderly]), aneurysmal location (middle cerebral artery territory [MC], anterior communicating artery [Acom] or internal carotid artery territory [IC]) and intraoperative trauma (positive or negative). We defined intraoperative trauma as intentional vein cutting, perforator vessel injury or suction of the gyrus rectus. All patients received standardized general anesthesia and were treated after the operations according to our standard protocols. Serum levels of CRP on postoperative day 2 were compared in each category. RESULTS: Serum levels of CRP showed no significant difference between the adult group and the elderly group, but were greater in Acom patients than in MC and IC patients (p < 0.01, p < 0.05). As to intraoperative traumas, the positive group (almost all were intentional vein cutting) showed greater CRP levels than the negative group in MC and IC patients (p < 0.05). Operation time and volume of blood loss were almost the same in each category. CONCLUSION: Our study shows that as far as serum levels of CRP are concerned, Acom location and vein cutting are the main factors of surgical stress in cases of unruptured aneurysms. PMID- 12533904 TI - [A case of subcutaneous malignant lymphoma with dura mater lesion]. AB - A 63-year-old male was admitted to our hospital, complaining of a scalp mass located at the frontoparietal area of his head. He noticed that it had been growing for 2 months. The mass was elastic hard and non-moving. Computed tomography demonstrated a subcutaneous mass with low density and which was enhanced homogeneously. The skull just below the mass was slightly destroyed, but the structure remained. Magnetic resonance imaging (MRI) demonstrated a mass with low signal intensity on both the T1 weighted image and the T2 weighted image. Gd DTPA study showed homogeneous enhancement and showed also that the dura just below the mass was enhanced. At this point we couldn't diagnose it confidently, but suspected this lesion to be a malignant lymphoma. We made a general examination, but no other lesion was found. A biopsy of the subcutaneous mass was performed under local anesthesia. The histological diagnosis was large-cell type B-Cell lymphoma. The tumor was treated with chemotherapy, CHOP (cyclophosphamid, doxorubicin, vincristin predonisolone). It responded to this chemotherapy and disappeared. We treated this lesion without radiation therapy. We report a case of subcutaneous malignant lymphoma treated successfully with a minimum invasive method. PMID- 12533905 TI - [A chronic intracerebral hematoma for which corticosteroid was effective: case report]. AB - Chronic intracerebral hematoma is rare. Computerized tomography almost consistently demonstrates ring-shaped lesions with mass effect and perifocal edema. All patients in published studies have been treated surgically. Most of the hematomas have been encapsulated and contained blood in various stages of organization. The thick capsule has usually consisted of an outer layer of collagenous tissue and an inner layer of granulation tissue. A 60-year-old male, who had had sudden neurological symptoms, left hemiparesis and aphasia, was admitted to our hospital. Fourteen days after admission, CT revealed an expanding low density mass with brain edema. He was treated surgically and with corticosteroid. In this case, the surgical treatment was not effective but the corticosteroid medication was effective. Strategies of treatment are discussed. PMID- 12533906 TI - [Diploic epidermoid cyst: case report]. AB - We report the case of a diploic epidermoid cyst in the left front-parietal bone of a 70-year-old woman. A skull x-ray showed an osteolytic lesion with a sclerotic border, and CT scan revealed a low/iso-density mass and the destruction of both inner and outer tables. MRI showed hypo-intensity on T1-weighted image, hyper-intensity on T2-weighted imaging and remarkably-strong intensity on the diffusion-weighted imaging. Gadolinium enhancement was seen in the tumor capsule and dura of the peripheral tumor. In the operative view, most of the bone edge was regular with a sclerotic border, but an irregular section was present. The tumor adhered strongly to the dura mater and periosteum. The tumor was totally extirpated with the peripheral skull, dura mater and periost. On the postoperative MRI, gadolinium enhancement of the nearby dura mater and disappeared. We have added a discussion of the radiological features of diploic epidermoid cysts. PMID- 12533907 TI - [Vascular reconstruction of tandem stenoses of the internal carotid artery]. AB - We report a case treated successfully by carotid endarterectomy combined with endovascular stenting for tandem stenoses of the internal carotid artery. A 71 year-old man had tandem stenotic lesions affecting bifurcation and the petrous portion of the left internal carotid artery. Coronary stents were placed at the petrous portion of the internal carotid artery immediately after standard carotid endarterectomy. Endvascular techniques and surgery can be complementary in the management of patients suffering from such tandem lesions. PMID- 12533908 TI - [A case of multiple mycotic intracranial aneurysms]. AB - A patient with multiple mycotic aneurysms associated with infective endocarditis is reported. A 45-year-old man was admitted on February 16, 2001 under the diagnosis of infective endocarditis. After alpha-streptococcus was identified by a blood culture, he was treated with high doses of antibiotics. However, 2 weeks after admission, he suddenly suffered from headache and mild left hemiparesis. A CT showed a parenchymal hematoma in the right parietal lobe. Cerebral angiography demonstrated aneurysms of the bilateral middle cerebral artery and the left posterior cerebral artery. At first, we trapped and resected the ruptured right middle cerebral aneurysm. After the surgery, we tried to treat two unruptured aneurysms by endovascular treatment. During the provocation test for the posterior cerebral artery, the arterial wall was perforated by a guide wire. The parent artery was occluded by coils at this site. Although the aneurysm was still filled by retrograde blood flow, it finally disappeared six months after treatment. The left middle cerebral artery aneurysm could not be treated because the provocation test showed cognitive deficits. The patient recovered from infective endocarditis after four-months of antibiotic therapy; and the unruptured aneurysm had not changed in size for 11 months. Recently, the outcome of patients with intracranial mycotic aneurysm is improved by development of multimodality management. Especially, endovascular therapy may become an effective treatment for unruptured aneurysms, but it is necessary to take risks, such as arterial perforation into consideration. PMID- 12533909 TI - [Degenerative disease (1), dementia, no. 7 in series of articles: basic knowledge of neuropathology for neurosurgeons]. PMID- 12533910 TI - [Problems and proposals from neurosurgical practice, no. 11, in series of articles: controversy and reform proposals concerning insured medical care in neurosurgery]. PMID- 12533911 TI - Comparative toxicity study of 3-aminophenol in newborn and young rats. AB - Repeated dose toxicity of 3-aminophenol was examined on oral administration to newborn and young rats, and susceptibility was analyzed in terms of the no observed adverse effect level (NOAEL) and the unequivocally toxic level. In the 18-day newborn rat study, starting at day 4 after birth, tremors and depression of body weight gain were observed, as well as hypertrophy of thyroid follicular epithelial cells and increases of relative liver and kidney weights at 240 mg/kg. Increase of relative liver weights in males and decrease of blood sugar in females without any histopathological changes at 80 mg/kg were not considered to be adverse effects. No chemical-related changes were observed at 24 mg/kg. Abnormalities of external development and reflex ontogeny in the newborn were not observed. In the 28-day study, starting at 5 weeks of age, depression of body weight gain, tremors, anemia, and liver, kidney and thyroid toxicity were observed at 720 mg/kg. Although slight pigmentation in the renal proximal tubular epithelium was observed in females at 240 mg/kg, this was not considered to be an adverse effect because of the lack of changes in related toxicological parameters. It was concluded that the NOAEL is 80 mg/kg/day in newborn rats and 240 mg/kg/day in young rats, with unequivocally toxic levels of 240 mg/kg/day and 720 mg/kg/day, respectively. Based on these two endpoints, the susceptibility of newborn rats to the chemical was approx. 3 times higher than that of young rats, consistent with our previous results for 4-nitrophenol and 2,4-dinitrophenol. PMID- 12533912 TI - Effects of kojic acid on thyroidal functions in rats by single-dose administration and in cultured rat thyroid cells (FRTL-5 cells). AB - The effects of kojic acid (KA) on thyroidal function were studied by single-dose administration in rats and in cultured rat thyroid cells (FRTL-5 cells). In rats receiving a single dose of 1,000 mg/kg KA orally, the 125I uptake from blood into the thyroid gland was significantly lower than that of the control group from 30 min to 24 hr after administration. The 125I organification activity of the KA groups was significantly lower than control from 30 min to 6 hr after administration. However, the 125I organification activity at 24 hr or 48 hr after administration recovered enough to be nearly comparable with the control group. In the study in FRTL-5 cells, KA inhibited iodine organification dose dependently, but did not inhibit iodine uptake. These results suggest that the observed lower iodine uptake activity in the single-dose administration study in rats was due to the inhibition of iodine organification caused by the oral administration of KA, consequently decreasing iodine in the entire thyroid gland. Although serum T4 showed a tendency to decrease from 2 hr to 48 hr after administration of KA, serum TSH did not show any evident change associated with KA in the single-dose administration study in rats. Based on these results, it is presumed that a massive dose or long administration period might be needed to decrease serum T4 and increase serum TSH. From these results, it is presumed that KA affected thyroidal function when given at a massive dose or in a long administration period by inhibiting iodine organification in the thyroid. PMID- 12533913 TI - Lack of significant alteration in the prostate or testis of F344 rat offspring after transplacental and lactational exposure to bisphenol A. AB - Bisphenol A (BPA), a compound of great concern as an estrogenic xenobiotic, was assessed for its ability to cause alteration in the accessory sex organs and spermatogenesis in male offspring exposed preneonatally and neonatally. In a series of experiments focusing on rat sensitivity to gestational and lactational exposure to BPA, we investigated its effects on gestation period and reproductive organs in male offspring. In the first instance, BPA was administered to F344 female rats by gavage at 0, 7.5, 120 mg/kg/day during pregnancy and lactation period. There were no observable adverse effects in pregnant rats and the treatment did not induce any morphological abnormalities in the accessory sex organs of male offspring. However, lowered numbers of sperm in the testis were found with a dose of 120 mg/kg/day. In the second study, the same protocol with a higher number of male offspring was applied, but no reduction in the sperm count was apparent. We conclude that transplacental and lactational exposure to BPA dose not exert any adverse effects on morphogenesis of rat accessory sex organs or spermatogenesis. PMID- 12533914 TI - A mechanism of the thearubigin fraction of black tea (Camellia sinensis) extract protecting against the effect of tetanus toxin. AB - The aim of the present study was to elucidate the mechanism of the protective effect of black tea extract's thearubigin fraction against the action of tetanus toxin. The effects of thearubigin fraction extracted from a black tea infusion were examined for neuromuscular blocking action on tetanus toxin in mouse phrenic nerve-diaphragm preparations and on the binding of this toxin to the synaptosomal membrane preparations of rat cerebral cortices. The interaction between tetanus toxin and thearubigin fraction was also investigated. Tetanus toxin (4 micrograms/ml) abolished indirect twitches in mouse phrenic nerve-diaphragm preparations within 150 min. Thearubigin fraction mixed with tetanus toxin blocked the inhibitory effect of the toxin. Mixing iodinated toxin with thearubigin fraction inhibited the specific binding of [125I]tetanus toxin to the synaptosomal membrane preparation. The effects of thearubigin fraction were dose dependent. The elution profile of [125I]tetanus toxin on Sephadex G-50 column chromatography was different from that of toxin mixed with thearubigin fraction. These findings indicate that thearubigin fraction protects against the action of tetanus toxin by binding with the toxin. PMID- 12533915 TI - Single or group housing altered hormonal physiology and affected pituitary and interstitial cell kinetics. AB - A significant negative correlation between testicular interstitial cell tumors and pituitary tumors in control male F344 rats has been reported associated with the number of animals per cage. Change in numbers of animals per cage may cause stress and increased serum corticosteroids that can impair testosterone synthesis by interstitial cells. Eventual atrophy of interstitial cells may result in pituitary hyperfunction and tumor development. For relevant risk assessments, understanding the effect husbandry has on cellular processes is necessary. Twenty four 6-week-old male F344 rats were housed individually, as pairs, or as trios for 13 weeks. Measured parameters included feed consumption, body and organ weights, hemograms, hormonal levels, histopathology, and cellular kinetics in the pituitary and testicle. Several caging-related differences occurred, that, although not statistically different, could be biologically significant; these included increased serum levels of estradiol, progesterone, and corticosterone; increased spermatogonial proliferation; decreased apoptosis within seminiferous tubules; and increased BrdU immunoreactivity of the interstitial cells. The statistically significant decrease in lymphocyte numbers correlated with the associated increase in corticosterone levels. This study indicates that the number of animals in a cage is associated with hormonal and cellular kinetic changes in the pituitary and testes, which could influence the incidence of tumors in these organs. PMID- 12533916 TI - [Genotoxicity studies of stevia extract and steviol by the comet assay]. AB - The genotoxicity of steviol, a metabolite of stevia extract, was evaluated for its genotoxic potential using the comet assay. In an in vitro study, steviol at 62.5, 125, 250, and 500 micrograms/ml did not damage the nuclear DNA of TK6 and WTK1 cells in the presence and absence of S9 mix. In vivo studies of steviol were conducted by two independent organizations. Mice were sacrificed 3 and 24 hr after one oral administration of steviol at 250, 500, 1000, and 2000 mg/kg. DNA damage in multiple mouse organs was measured by the comet assay as modified by us. After oral treatment, stomach, colon, liver, kidney and testis DNA were not damaged. The in vivo genotoxicity of stevia extract was also evaluated for its genotoxic potential using the comet assay. Mice were sacrificed 3 and 24 hr after oral administration of stevia extract at 250, 500, 1000, and 2000 mg/kg. Stomach, colon and liver DNA were not damaged. As all studies showed negative responses, stevia extract and steviol are concluded to not have DNA-damaging activity in cultured cells and mouse organs. PMID- 12533917 TI - [Methane emission from rice paddy soils as influenced by soil physicochemical properties]. AB - To identify the key soil parameters influencing methane emission from rice paddies, and to quantitatively describe the relationship of methane emission with the soil properties, an outdoor pot experiment with a total of 18 paddy soils sampled from different regions in Jiangsu province was conducted in Nanjing Agricultural University during 2000 rice growing season. Seasonal average rate of CH4 emission for all the 18 soils was 6.42 +/- 2.70 mg.(m2.h)-1, ranging from 1.96 to 11.06 mg.(m2.h)-1, approximately a 5.6-fold difference between the maximum and the minimum. Correlation analysis indicated that the seasonal average of CH4 emission was positively dependent on soil sand content (r = 0.528, p = 0.024) and negatively on soil clay content (r = -0.484, p = 0.042). Negative correlation of CH4 emission against soil total nitrogen (r = -0.449, p = 0.062), available nitrogen (r = -0.611, p = 0.007) and NH4(+)-N(r = -0.649, p = 0.004) was also observed. Copper content of soils has a significant negative impact on CH4 emission. The correlation coefficient (r) of CH4 emission against soil available copper and with total copper was -0.594 (p = 0.009) and -0.547 (p = 0.019), respectively. No clear relationship existed between CH4 emission and soil carbon content. A further investigation suggests that the seasonal average rate of CH4 emission can be quantitatively determined by a linear combination of soil available copper, total manganese, and a ratio of available to total iron (R2 = 0.755, p = 0.0001). PMID- 12533918 TI - [Important factors controlling rates of N2O emission and CH4 oxidation from forest soil]. AB - In this study, laboratory incubation experiments were conducted to determine the effect of pH, soil water content, temperature, NH4+ and NO3- on N2O emission and CH4 oxidation of mixed broad-leaved Korean pine forest soil. Results showed that there were significant correlations between N2O emission rate, CH4 oxidation rate and soil pH and temperature under certain lab conditions. Meanwhile, the significant positive correlation between N2O emission rate and CH4 oxidation rate was also observed. The results suggested temperature and pH could be the most important factors controlling the rates of N2O emission and CH4 oxidation of a forest soil under this experiment conditions. PMID- 12533919 TI - [Use of biological activated carbon to degrade benzene and toluene in a biofilter]. AB - The biodegradation of toluene and benzene in a biofilter using cylindrical activated carbon as the filler materials was studied. For total mass loading lower than 350 g/(h.m3), retention time ranging from 15 s to 90 s the biofilter proved to be highly efficient in biodegradation of toluene and benzene, their removal capacity were 120 and 150 g/(h.m3) respectively and toluene was more easily degraded than benzene. The CO2 produced increased with the degradation of benzene and toluene, but the experimental value was lower than the theoretical value. The observation of biotic community demonstrated that the microbes consisted of bacillus, spore bacillus and fungi, of them spore bacillus was dominant. According to basic theories of mass-transfer, adsorption and biodegradation process, mathematical model of removal of VOCs by biofiltration was established and verified. PMID- 12533920 TI - [A backward trajectory inversion model for methane emission over Beijing area]. AB - A Backward trajectory inversion model was established for estimating methane emission over Beijing area, with in situ data of methane measurement. The model was systematically tested for its ability on reflecting diffusion processes in atmospheric boundary layers. In specific condition, the model acted excellently on calculating emission strength and distribution over an area of 100 km scale. By applying to Beijing area with 9 days real methane data and practice meteorological data, the model showed source patches in this area with emission rate 0.0066-0.026 mg/(m2.s), which was correspond well to rice fields emission, the major methane source, in this region. Model results of source patches also showed certain spatial correlation with rice fields distribution. PMID- 12533921 TI - [Calculating critical loads of acid deposition with different percentiles in China]. AB - While mapping critical loads of acid deposition in China, the 1 degree (latitude) x 1 degree (longitude) resolution was always adopted in critical load calculation. However, the results of mapping can not show the difference of sensitivity of ecosystems to acid deposition within a 1 degree x 1 degree grid. For the convenience of policy-makers to formulate acid deposition control strategies based on critical loads, and to improve the representation and practicability of 1 degree x 1 degree results, a series of critical load maps with different percentiles were compiled, which may be accordance with a given economic or technological level, and allows some degree of damage. Based on the cumulative distribution function, the critical load exceedance maps with different percentiles and the maximum allowable deposition of each province was also derived. PMID- 12533922 TI - [Relationship between population variation of fig trees and environment in the tropical rainforests of Xishuangbanna]. AB - The species diversity of the plant plays an important role in the ecological environment conservation. In the tropical rainforest of Xishuangbanna the biological diversity is extremely rich, and which is a key region used to biological diversity research. And the fig is a key plant species in the tropical rainforest. The research results about the relationship between the variation of the figs population and environment of the region were following: 1. There were 69 species, variants and subspecies Ficus plant that was known in the tropical rainforest of Xishuangbanna, thereinto there were 23 species and subspecies owned to the Urostigma, and the Pharmacosyea had four species; the Sycomorrus two species and subspecies; while the Ficus owned 41 species, subspecies and variants, then Xishuangbanna borne the most richness and diversity of fig species and numbers in China. 2. The distribution of the figs had intimate connection with the environment, and the species and the quantities showed the most richness in the tropical primary rainforest that was protected well and almost intact, where the figs were mainly to be arbors and giant arbors; and in the devastated stand and secondary forest dwarf arbors, shrubs, bushes and ligneous liana possessed the most in the fig species; while in such regions of tropical rainforest environment that have been destroyed seriously one proportion of the pioneer species of the figs were the main members. 3. Only the key colony of Ficus can be conserved in the tropical rainforest, can the whole ecological environment of the tropical rainforest be protected and rehabilitated, which was one of the important measures to realize it. PMID- 12533923 TI - [Runoff phosphorus discharge from different land use system in subtropical hilly areas]. AB - The environmental effect of land use becomes one of the most important issues in ecosystem research. This study on phosphorus in runoff was conducted in two contiguous sub-watersheds with three land use types, which located in Meicun watershed of Xuancheng County, Anhui Province. The sub-watersheds differed in land use, but had similar soils and hydrology. The results showed that the runoff dissolved phosphorus (DP) and particulate-associated phosphorus (PAP) concentration of forest-pond-paddy-village system was the highest among the three land use systems. There was no significant difference between DP of the forest pond system and forest-paddy system. However, the PAP and TP of forest-paddy system was higher than that of forest-pond. The rank of the DP, PAP and TP discharges was wholly same with that of the PAP concentration. About 90% of TP was discharged with the particulate soil. There was a significant linear relationship between the concentration of PAP and the particulate of runoff in three land use systems. But only in the forest-pond-paddy-village system existed a logarithm linear relationship between DP and PAP concentration. PMID- 12533924 TI - [Stress effects of simulant acid rain on three woody plants]. AB - Osmanthus fragrana, Chimonanthus praecox and Prunus persica were used as materials to investigate the effect of simulant acid rain on chlorophyll (Chl) content, cell membrane permeability(L%), the content of proline (Pro) and malondialdehyde (MDA) in three woody plants with different resistance, and effects of the light and dark conditions on acid rain injury. The results showed that the change degree of four kinds of physiological and biochemical indexes for these woody plants was as sequence: Osmanthus fragrana > Chimonanthus praecox > Prunus persica. The change of chlorophyll content in these woody plants was not obviously when acid rain stress was influenced by the light and dark. PMID- 12533925 TI - [Sub-acute hepatotoxicity of low doses of microcystins]. AB - In order to study sub-acute hepatotoxicity of low doses of microcystins in vivo, as well as to understand the mechanisms of hepatotoxicity of microcystins, eighty Spague-Dawley rats were injected with microcystins intraperitoneally at the doses of 0, 4, 8 and 12 micrograms.(kg.d)-1, respectively, for 35 days. Then blood and liver samples were used for assay. Several enzymatic levels and pathological changes were detected. Both terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) and immunohistochemical methods were employed to study the apoptosis and proliferating cell nuclear antigen (PCNA). It was shown that the activity of serum gamma-glutamyltransferase (GGT) and concentrations of whole blood glutathione (GSH) decreased, serum activities of lactate dehydrogenase (LDH) and aminotransferase (AST) increased after exposure to MC. No significant change of concentration of serum alanine aminotransferase (ALT) was observed in the tested groups. Characteristic morphological alterations and active proliferation as well as apoptosis of hepatotocytes were observed in the treated groups. It was suggested that oxidative injury and apoptosis of hepatocytes induced by microcystins may be the mechanisms of its hepatotoxicity. PMID- 12533926 TI - [Effect of sewage sludge and its composts on the accumulation of organic pollutants in Ipomoea aquatic grown in paddy soil]. AB - Eight kinds of organic pollutants including 44 compounds in Ipomoea aquatic grown in paddy soil fertilized with sewage sludge and its composts were systematically determined by GC/MS. The results showed that twenty eight compounds consisting mainly of phthalic acid esters, nitrobenzenes, and polycyclic aromatic hydrocarbons were deteced in Ipomoea aquatic. Only individual or very few compounds for each kind of organic pollutants in Ipomoea aquatic were predominant, while others were trace or not detected. Compared with control, compounds detected in Ipomoea aquatic fertilized with sludge were commonly higher to a different extent, but those detected in Ipomoea aquatic fertilized with different sludge composts were commonly lower and even not detected, especially for Ipomoea aquatic fertilized with sludge + rice straw composts was detected only one compound. It was therefore concluded that direct use of sewage sludge as organic fertilizer could result in bioaccumulation of organic pollutants in crops, but sludge compost application would be safe if proper compost treatment for sludge was conducted. PMID- 12533927 TI - [Disinfection and degradation of 2,4-DCP with UV-radiation and on-line ozone in drinking water treatmeant]. AB - A type reactor with on-line O3 was used to do further research of the disinfection of total bacteria, E. coli and degradation of 2,4-DCP. The result was obtained in the following conditions. Only UV-radiation, O3 applied by other machine and by the reactor itself, and other conditions were changed to study the disinfection and degradation. The result showed the satisfied effect of disinfection and degradation would be achieved by using UV/O3 applied outside and when the flow rate was about 400 L.h-1, on-line O3 could be produced and make high efforts to enhance disinfection and degradation. The method of UV/O3 was a promising technology in the treatment of drinking water. PMID- 12533928 TI - [Nitrification of high concentration NH4(+)-N in membrane bioreactor]. AB - Nitrification performance of a membrane bioreactor was investigated under high concentration of NH4(+)-N. The removal rate of NH4(+)-N was over 99% under the condition of influent NH4(+)-N of 2000 mg/L and volumetric loading rate of NH4(+) N 2.0 kg/(m3.d). The specific nitrification rate was kept to be 0.36/d in a period of half year. PMID- 12533929 TI - [Study on removal of nitrobenzene in water by O3/H2O2]. AB - Comparative experiments of ozonation and O3/H2O2 processes were conducted with nitrobenzene as the model organic pollutant. It was found that O3/H2O2 greatly increased the removal efficiency of nitrobenzene compared with that achieved by ozonation alone. It was believed that both in ozonation and O3/H2O2 processes nitrobenzene was primarily oxidized by OH.. Several factors that are likely to influence O3/H2O2 process in practical application were investigated. Experimental results demonstrated that with the same dosage of applied ozone and hydrogen peroxide, the multiple steps addition of ozone and hydrogen peroxide showed a much higher removal efficiency than that obtained by adding hydrogen peroxide within one step before the start of the experiment. Under the conditions of this experiment, the optimum molar ratios of hydrogen peroxide to ozone for the removal of nitrobenzene in distilled water and tap water were proved to be both 0.5, no significant negative effect was observed when the alkalinity levels (as CaCO3) were less than 100 mg/L. PMID- 12533930 TI - [Treatment of drilling wastewater from oil field by using yeast]. AB - Two strains of yeast, namely Wickerhamiella domercqii and Candida boidinii, were acquired through screening from soil samples contaminated by drilling wastewater. A TOC removal of 40.5% was acquired when the mixture of the two yeast strains was used for drilling wastewater treatment, a little higher than that with activated sludge acclimated with wastewater (35.2%). Some organic compounds in the fraction of molecular weight above 60,000 were found to be biodegradable. PMID- 12533931 TI - [Biodegraded characteristics of MGP-wastewater by domestic and screened organisms]. AB - Among 16 bacteria that were screened out from 5 activated sludge, S-2, Y-3, XH-3 and their mixed strains M-3 were selected to biodegrade MGP (Manufacture gas from petroleum)-wastewater. They can remove 38.4%-44.0% CODCr, 95.4%-97.0% hydroxybenzene and 47.1%-53.7% aromatic compound after 60 h treatment, but the removal rate of NH4(+)-N was only 14.0%-17.6%. About 77 toxic organic compounds were detected by GC/MS analysis, the aromatic compounds which had 3 rings and 4-6 rings can be biodegraded by the bacteria. The removal rate of CODCr, NH4(+)-N and extractable organic compound were improved 13.1%-22.9%, 18.4%-22.7% and 13.1% 18.1% respectively when glucose was added in the wastewater, and when ethanol was existing in the wastewater, the corresponding increase were 12.5%-21.2%, 29.7% 42.2% and 6.7%-7.7%. The total removal rates of aromatic compound were increased 17.7%-21.7%, 15.4%-21.2%. The screened strains can improve the biodegradation rate of MGP-wastewater. PMID- 12533932 TI - [The transformation of nonylphenol ethoxylates in sewage treatment]. AB - The concentrations of individual NPnEO (n > 2) were separated and analyzed by normal phase liquid chromatograph-mass spectrometry (NPLC-MS), and those of NP, NP1EO and NP2EO were determined by GC-MS. NP and NPnEO (n = 1-23) were found in the influent water, and the concentration of total NPnEO was 174 nmol/L. Of NPnEO, the abundance of NP2EO (89.6 nmol/L) was the highest, and next was NP (42.2 nmol/L) followed by NP1EO (12.2 nmol/L). 71% NP and NPnEO were removed by sewage treatment process and the adsorption on sludge contributed to the removal for these compounds. It was found that the adsorption of NPnEO on sludge followed the Dubinin-Astakhov model, assuming each NPEO were with similar adsorption potential. PMID- 12533933 TI - [The kinetics of biological treatment of bleaching liquor from straw pulp process]. AB - The batch test of oxygen uptake rate (OUR) measurement was used to simulate biochemical reaction process in aerobic reactor treating bleaching liquor from straw pulp process. Lawrence-McCarty kinetic model was used to analyze the biochemical reaction process. The kinetic equation of bleaching liquor biological treatment, v = 0.72 S/(60.43 + S), was deduced from results of the OUR experiment and the bench experiment. The specific substrate degradation rate estimated by the kinetic equation was higher than that of practical wastewater treatment plant. The phenomenon was attributed to the decrease of activated sludge activity caused by cellulose suspended solid accumulation in aerobic reactor of practical plant. PMID- 12533934 TI - [Removal of arsenate by a new type of ion exchange fiber]. AB - A new type of ion exchange fiber was prepared for the removal of arsenate from water. A batch sorption experiments showed that the fibrous sorbent had high sorption capacity and good kinetic property for arsenate ion. The sorption kinetic data can be described by the Lagergren pseudo-second order rate equation very well. Freundlich model can simulate the adsorption equilibrium data of arsenate ion in the studied concentration range. The removal of arsenate by the fiber reached a maximum in pH value range of 3.5 to 7.0. The column performance was carried to assess the applicability of the ion exchange fiber for the removal of arsenate ion with satisfactory result. The diluted NaOH solution is an efficient eluant for the desorption of arsenate on from the fiber column, and 30 mL of 0.5 mol/L NaOH can quantitatively recover arsenate from water. PMID- 12533935 TI - [Experimental study on prediction model for leachate quality variation in MSW landfills]. AB - The changing rule of leachate quality in Xiaping Sanitary Landfill was studied using pilot-scale leaching test for waste pollutants. And the parameters for Pollutants Leaching Model were determined using the data from the test by exponential regression method. Both the results from the test and the monitoring data in site indicated that the leachate concentration reached its maximum with COD of 31,581 mg/L after 75 days. Meanwhile, the accuracy of this prediction model was testified by comparing with the monitoring data in site, that means the model and its parameters achieved by leaching test for pollutants load can be used to predict the leachate quality variation in MSW landfills. Thus it provides a scientific numerical model and leaching test method for leachate quality prediction and basis for designing and managing leachate treatment facilities. PMID- 12533936 TI - [Horizontal transport of nitrate in main soil groups of Huang-Huai-Hai Plain]. AB - The horizontal transport of nitrate in main soil groups (yellow fluvo-aquic soils and aeolian sandy soil) in Huang-Huai-Hai Plain was studied. The results were as follows: the horizontal transport velocity of nitrate decreased with the distance of the tracer source in power function. Due to different soil property in soil profile, the curves of horizontal transport of nitrate were difference. The horizontal transport velocity of nitrate was controlled by the concentration gradient and soil water potential gradient in 20 cm of horizontal soil column. It was stable after 20 cm, which was controlled by soil matric potential. The horizontal transport velocity of nitrate was in a sharp positive relation with the soil moisture content and changed with exponential function. The concentration of nitrate in horizontal transport decreased with the unsaturated soil water diffusivity increased and changed with logarithmic function. PMID- 12533937 TI - [Transformation of Na, K, Pb and Mn during pyrolysis of coal]. AB - Transformation of Na, K, Pb and Mn in char for Shenhua and Yima coals at pyrolysis temperatures of 300 degrees C-700 degrees C was reported. Effects of atmosphere (N2, H2 and CO2) and pressure on release of the elements in Shenhua coal were also studied. The pyrolysis experiment was carried out in a closed stainless steal reactor. Results show that the bleeding ratio of Na, K, Pb and Mn varied with coal type but all increased with pyrolysis temperature. For example, the bleeding ratio of Na, K, Pb and Mn at 700 degrees C for Shenhua coal was 3, 4.2, 1.4, and 2.4 times larger than that of Yima coal. The effect of pressure was small. Among the N2, H2 and CO2 environments, H2 resulted in the highest bleeding ratio and CO2 the least bleeding ratio. Thermodynamic calculation shows that Pb was the most volatile element among the four inorganic harmful elements. PMID- 12533938 TI - [Sodium-enhanced limestone wet FGD in rotating-stream tray scrubber]. AB - Adding sodium sulfate to limestone slurry can increase SO2 removal efficiency. In this paper, sodium-enhanced limestone flue gas desulfurization(FGD) tests were conducted in rotating-stream tray scrubber. Changes of SO2 removal efficiency and pH value with time were experimentally studied and, at different pH range, the dissolution rates of limestone etc. were analyzed. The effects of plate number on SO2 removal efficiency and pressure drop were investigated at temperature approximated to industrial operating value. The average plate efficiencies were calculated. According to the experimental results, increasing plate number could increase SO2 removal efficiency, but the average plate efficiency decreased. Under the experimental conditions, when the plate number was increased from 1 to 4, the removal efficiency was increased from 25.5% to 48.6% at the liquid-to-gas ratio of 4 L/m3, but the average plate efficiency was decreased from 25.5% to 15.3%. The equation of the relation between the average plate efficiency and plate number was obtained. PMID- 12533939 TI - [Adsorption of Cd and Pb on clays and the influence of humic acids]. AB - Adsorption of cadmium and lead on montmorillonite and kaolin, together with the effect of humic substances was studied. It was found that adsorption amount of Cd and Pb increased as pH value of ambient solution increased. Addition of FA reduced Cd sorption on kaolin at each respective pH value while GHA and BHA increased Cd adsorption amount. A sharp increase was found at pH values higher than 6. Whereas, lead sorption on both clays in the presence of FA unexpectedly decreased as pH values increased when pH > 6. PMID- 12533940 TI - [Extraction of organochlorine pesticides using ASE from wastewater irrigated agricultural soil]. AB - The extraction efficiency of accelerated solvent extraction (ASE) was studied for organochlorine pesticide residuals in soil and compared with that of Soxhlet. The results showed that the efficiency of ASE was generally better than Soxhlet when used for DDT measurement, and equivalent to Soxhlet for BHCs. With ASE, organochlorine pesticide residues were detected in wastewater irrigated and non wastewater irrigated soils from Tianjin. alpha-BHC, beta-BHC, delta-BHC, gamma BHC, p,p'-DDE, p,p'-DDD, p,p'-DDT, o,p'-DDT ranged from 7.5 to 71.1 ng/g in wastewater irrigated vegetable and maize field and from 3.0 to 16.5 ng/g in wastewater irrigated paddy field. The eight pesticide residues from non wastewater irrigated vegetable and maize field was in a range of 3.1-17.6 ng/g. PMID- 12533941 TI - [Determination of total arsenic in sinters by hydride generation atomic fluorescence spectrometry]. AB - A simple and sensitive method was proposed for the determination of total arsenic in sinters by hydride generation atomic fluorescence spectrometry (HGAFS). The method was based on the oxidation of arsenic compounds with strong acid mixtures in the pressure steel bomb. The linear range for AsV determination was 0-300 micrograms/L. The method was validated by analysis of standard reference material (coal fly ash) with satisfactory results and was successfully applied to the determination of total arsenic in sinters with RSD less than 4%. PMID- 12533942 TI - [Cost-benefit analysis to substituting natural gas for coal project in large Chinese cities]. AB - Since China's large cities were faced with serious coal-smoke pollution with PM10 and SO2 as the main pollutants, natural gas is becoming one of the most attractive clean replacers of coal. To clarify the wide disputation and doubt on the rationality of burning natural gas instead of coal, cost-benefit analysis (CBA) of urban natural gas substitution projects in Beijing and Chongqing was done respectively, in which, the health benefit was carefully estimated with epidemical dose-response function as the main external benefit. The final result shows that in large cities with intensively concentrated population and economic activities, natural gas consumption as municipal civil energy has obvious priority in terms of large environmental benefit from reducing non-point and low altitude air pollutant concentration. This paper finally recommends that market oriented system reform in natural gas production and retailing system should be considered. PMID- 12533943 TI - [Effect of triadimefon on activity of peroxidase and soluble protein content in detached cucumber cotyledons]. AB - Contents of chlorophyll a, chlorophyll b, soluble protein and proline, as well as the ratio chlorophyll a/b in detached cotyledons of cucumber were distinctly increased by treating with 20 mg/L triadimefon, which suggested that triadimefon can effectively retard the senescence of detached cotyledons in cucumber. Triadimefon could inhibit the increase of peroxide (POD) activity and H2O2 content in different degree. It was found that contents of chlorophyll and protein were markedly negative correlated with the activity of POD and with the content of H2O2 respectively during the senescence of cotyledons. These results indicated triadimefon inhibit the degradation of chlorophyll and protein in cucumber cotyledons through POD- H2O2 enzyme system. PMID- 12533944 TI - Selected local and peripheral intereactions of some cytokines in etiopathogenesis of preeclampsia. AB - In accordance with presented results and clinical observation it should be assumed that basic phenomena resulting in clinical sign of preeclampsia take place already in implantation and embryogenesis. Abnormal trophoblastic infiltrations and changes in vascular endothelium functioning occur due to disturbed interaction of cytokines and growth factors. Pathologies present in endothelial cells (EC) become the cause of blood overcoagulation and hypertension and promote other clinical symptoms of preeclampsia. PMID- 12533945 TI - Endocrine and intermediate cells in pancreatic ductal carcinoma and in chronic pancreatitis. AB - As most pancreatic ductal carcinomas have the genotype of ductal epithelial cells it is believed that they originate from the epithelium of these structures. It has been proved that endocrine cells are frequently present in ductal carcinoma. Morphological, immunohistochemical, ultrastructural evaluation and statistical analysis revealed the presence of one or a few types of endocrine cells in 46.9% of ductal carcinomas (mainly of I degree or II degree malignancy). These cells were found to secrete insulin. Intermediate cells were not observed in ductal carcinoma. The structure of chronic pancreatitis, mainly the so called tubular complexes, showed various types of endocrine cells and numerous intermediate cells, often with features of destruction. The present study seems to prove that pancreatic cells have a common origin-acinar cells as well as islet cells arise from ductular cells. The intermediate cells observed in chronic pancreatitis are not just a transitory form in the differentiation of precursor cells, but rather the abnormal forms of cells that are produced in that process. PMID- 12533946 TI - Effect of high-fat diet on sarcoplasmic reticulum Ca(++)-ATPase activity in different types of rat skeletal muscle. AB - The aim of the present study was to examine the effect of a high-fat diet on the Ca(2+)-ATPase activity in the sarcoplasmic reticulum of the flexor digitorum longus (fast-twitch, glycolytic muscle) and the soleus (slow-twitch, oxidative muscle). The enzyme activity in the extensor digitorum longus was several-fold higher than in the soleus. The high-fat diet increased the enzyme activity by 22% (p < 0.05) in the soleus but it had no effect in the flexor digitorum longus. It is concluded that a high-fat diet increases the activity of only one isoform of the enzyme, namely the one which is present in the slow-twitch oxidative fibers. PMID- 12533947 TI - Zoosporic and conidial fungi within the Podlasie stretch of the river bug. AB - Zoosporic and conidial fungi species and environmental factors in 5 various sites of the Podlasie stretch of the river Bug were studied. Samples of water were collected in October 1999 and April 2000 for hydrochemical analysis and the order to determine the number of fungal species. Buckwheat-seeds, hemp-seeds, cellophane and snake exuviae were used as bait during the exposure in laboratory. Sixty-four zoosporic fungus species and forty-six conidial species were found in various sites of the Podlasie stretch of the river Bug. 8 species were recorded for the first time from Poland. PMID- 12533948 TI - Further studies on aquatic fungi in the River Narew within the Narew National Park. AB - Zoosporic and conidial fungi species and environmental factors in 10 various sites of River Narew in the Narew National Park were studied. Samples of water were collected in autumn 1999 and spring 2000 for hydrochemical analysis and the order to determine the number of fungal species. Buckwheat-seeds, hemp-seeds, cellophane and snake exuviae were used as bait during the exposure in laboratory. 198 lower aquatic fungus species were found in various sites of River Narew. 13 species were recorded for the first time from Poland. PMID- 12533949 TI - Serum hyaluronan as a possible biomarker of liver insufficiency in cirrhotic patients. AB - Hepatic hyaluronan is a glycosaminoglycan synthesised in the liver mainly by mesenchymal cells and degraded by liver sinusoidal endothelial cells. Serum hyaluronan was measured, using enzyme-linked binding protein assay (Chugai, Japan) in 30 patients with cirrhosis of different etiologies (alcohol n = 15, primary biliary cirrhosis n = 4, hepatitis B n = 5, hepatitis C n = 6). Hyaluronan levels were significantly higher in patients with liver cirrhosis (398 +/- 375.4 ng/ml) than in the controls (36.5 +/- 26.9 ng/ml). There was no significant difference in hialuronan levels between the different etiologies of the disease. Hyaluronan level increased proportionally to the severity of cirrhosis. PMID- 12533950 TI - The formation of oxidative stress condition in the experimental chemically induced hepatotoxicity. AB - In spite of extensive studies concerning the chemically induced hepaototoxicity, no clear-cut practical conclusions were obtained. Wide chemical structure of compounds as well as the different way of administration are the most important factors. We used an acute administration of carbon tetrachloride and paracetamol and a chronic intoxication with acetylphenylhydrazine, cadmium chloride and ethanol. For the acute administration of chemicals in high doses, in the rat liver microsomes a decrease of cytochrome P450, aniline hydroxylase and glutathione (GSH) was noticed with an increase of lipid peroxides even at 2 hours after administration. 24 hours after the chemicals administration, in the blood of rats characteristic changes of hepatotoxicity such as increases glutamate pyruvate transminase, triglycerides and lipid peroxides, while GSH was decreased. In chronic administration, the cummulated concentration of chemical was true main factor. For both ways of chemicals administration, as a critical concentration was reached, an oxidative stress was produced, demonstrated by the increase of lipid peroxides and subsequently decrease of GSH. In both ways of administration, the oxidative stress was the precocious biochemical event occurring even at 2 hours and it triggers other metabolic changes which favor fatty infiltration and liver damage. PMID- 12533951 TI - The influence of cervical lesion on the stress state and strength of tooth with occlusal restoration: a numerical model study. AB - This paper is concerned with bio-engineering problems in dentistry. The analysis of stress in a lower premolar with a deep non-carious cervical lesion and restored occlusal Class I cavity under occlusal load of 100 N was shown. Finite element method was used for the analysis of a two-dimensional model of a human tooth. In order to facilitate the evaluation of the loads, four cross-sections in characteristical points of the model were made. The model was generated and the calculations were done on IBM Pentium III personal computer. The principal and equivalent stresses were calculated and used in qualitative and quantative evaluation of tooth tissue and tooth restoration strength. The results are shown in figures and charts. The analysis of the cross-sections distant to the cervical area has proved that the presence or absence of cervical lesion has no significant influence on the values and stress distribution. Only in the region of sharp-shaped notch in the bottom of defect (4-4) high stresses in the tooth tissues occur. The values of stress did not exceed the compressive and tensile strength in any case under investigation. The study has shown that the presence of cervical angular lesion does not affect the mechanical strength of a tooth with a properly restored occlusal cavity. Clinically, the situation is safe from the biomechanical point of view. PMID- 12533952 TI - Lymphocyte subpopulations in hypertrophied adenoid in children with otitis media with effusion. AB - Otitis media with effusion (OME) usually coexist with the adenoid hypertrophy. The aim of this study was a quantitative evaluation of the lymphocyte subpopulations in hypertrophied adenoid in children with persistent (ove three months) otitis media with effusion. We used flow cytometry for study the subpopulations of the lymphocytes in the pharyngeal adenoid during otitis media in the children, alternative group consist of children with simple adenoid hypertrophy. The monoclonal antibodies selected were specific markers for T lymphocytes (CD3), T-helper cells (CD4), T-suppressor cells (CD8), B cells (CD19) and NK cells (CD56). There were also analyzed T activation markers: HLA-DR and CD25. The percentage of NK cells was significantly decreased in the group of younger children with otitis media with effusion (0.81 +/- 0.51) in comparison to older ones (1.40 +/- 0.59%). Also, in the group of younger children with adenoid hypertrophy, the NK cells percentage was significantly lower (0.85 +/- 0.37%) than in the group of older children--(1.46 +/- 0.83). In both subgroups of younger children the percentage of lymphocytes T with HLA-DR activation marker was close, but it was significantly lower in the group of older children with otitis media with effusion (4.28 +/- 3.40%) in comparison to the group of older children with adenoid hypertrophy (5.03 +/- 3.00%). PMID- 12533953 TI - The decrease in antioxidant potential in human brain tumours. AB - The objective of our research was to estimate the activity of superoxide dismutase (SOD-1), glutathione peroxidase (GSH-Px) and glutathione reductase (GSSG-R), as well as the concentrations of free oxygen radicals "sweepers" (GSH, Vit.C) and the concentrations of the substances reacting with thiobarbituric acid in brain tumours with II, III and IV level of biological malignancy. The research was conducted on 61 samples obtained from the patients operated in the Department of Neurosurgery of the Medical Academy of Bialystok between the years 1996 and 2000. The obtained values of the above enzyme activities and of the concentrations of the examined substances in brain tumours were compared to the adequate values determined in 10 samples of histopathologically unchanged nerve tissue. THE RESULTS: the increase of all enzymes activities in brain tumours is statistically significant (p < 0.05) as compared to the adequate activity level in normal nerve tissue. The significant decrease (p < 0.05) of the GSH and ascorbate concentration and the significant increase (p < 0.05) of the level of the thiobarbituric acid--reactive substances in the examined brain tumours was found in comparison to the adequate concentration of the above substances in normal nerve tissue. PMID- 12533954 TI - Coenzyme Q10 level in plasma of children with inflammatory process. AB - Using a newly developed method by HPLC with ultraviolet detection we measured plasma coenzyme Q10 (CoQ10) level in group of 43 children (19 females and 24 males: ages 1 month-9 years) with an acute inflammatory process. The results for coenzyme Q10 were expressed as molar concentration (mumol/l plasma). Our study confirmed that CoQ10 concentration (median--0.8 mumol/l) was independent of sex, and we established which biochemical parameters influence on ubiquinone levels. The results indicate that CoQ10 concentration is connected with leukocytosis, calcium and magnesium levels. These findings suggested that transferin, amylase and serum glutamic transmainase may also determine the CoQ10 plasma levels. PMID- 12533955 TI - Analysis of the etiology, clinical picture and treatment of epilepsy in children in the north-eastern region of Poland. AB - Epilepsy is one of the most frequent diseases of the central nervous system (CNS) in children. It is considered that roughly 50% of epilepsies begin during childhood. The Department of Childrens' Neurology of the Medical Academy of Bialystok embraces with medical attention over 500 children with epilepsy from the north-eastern region of Poland. The aim of the study was the analysis of etiology, clinical symptoms and treatment of epilepsy in children, who are under the medical attention of our Department. 392 children aged from 7 months to 18 and 2/12 years were included in the study. In the analysed groups were 46.6% girls and 53.4% boys. 44.4% of the children live in cities and 55.6% in small towns or villages. The etiology of epilepsy was established in 48.7% of the children. The most common etiology of epilepsy were hypoxic-ischemic damages in perinatal period, developmental malformation of the brain, craniocerebral injuries and neuroinfections in the neonatal period. In 50.3% of the children the etiology of epilepsy was unknown. The most common types of seizures were primary and secondary generalised tonic-clonic seizures. 92.6% of the patients received the antiepileptic drugs during the study. Monotherapy was administered in 66.0%, politherapy in 26.5% of the children. The results relating to the etiology and clinical picture of epilepsy in our children are consisted with many dates from literature. It is a favourable observation, that in many children monotherapy is an effective treatment. PMID- 12533956 TI - Chronic neurotic and depressive disorders as posttraumatic reactions in the Polish Siberians. AB - The study "Chronic depressive and neurotic disorders as posttraumatic reactions in the Polish Siberians" focuses on the effect of traumatic factors in the form of chronic distress on mental and physical state. Test results confirmed earlier clinical observations of depressive and neurotic disorders, and the Posttraumatic Stress Disorder (PTSD). PTSD is a reaction associated with an event a threat to life or to physical integrity, during with a person experience sever distress. PMID- 12533957 TI - Effect of diabetes and contractile activity on incorporation of the plasma-borne fatty acids into skeletal muscle lipids. AB - There are only few data on skeletal muscle lipid metabolism in diabetes. The aim of the present study was to examine effect of streptozotocin diabetes on incorporation of the blood-borne 14C-palmitic acid into different fractions of skeletal muscle lipids at rest and during contractile activity. The experiments were carried out on male Wistar rats. Streptozotocin was administered intravenously. The control rats received saline. Seven days later, the rats were anaesthetized and the calf muscles of one hindleg were made to contract for 1 min by stimulation of the sciatic nerve with tetanic pulses (30 tetani/min). Thereafter, 14C-palmitic acid was administered intravenously and the stimulation was continued for 5 min. The contralateral resting leg served as a control. Samples of the soleus, red and white gastrocnemius were taken. Lipids were extracted with chloroform/methanol and separated into the following fractions by means of thin layer chromatography: phospholipids, mono-, di-, and triacylglycerols, free fatty acids, cholesterol and cholesterol esters. Radioactivity of each fraction was counted. It has been found, that the label was always incorporated mostly into the fraction of triacylglycerols and phospholipids. Diabetes increased radioactivity of each lipid fraction, both at rest and during contractile activity, comparing to the respective values in the control rats. It is concluded that transport of the blood-borne free fatty acids into the myocytes and their incorporation into different lipid fraction increases in acute diabetes. PMID- 12533958 TI - Fetotoxic action of 2,4-dichlorophenoxyacetic acid (2,4-D). III. Morphological changes in rat kidneys. AB - The paper presents the findings of histological studies on the effect of sodium salt of 2,4-D acid on the changes within kidneys in newborn rats exposed to this herbicide in the prenatal and postnatal period. The experiment was performed on 60 Wistar rats of both sexes, up to 10 weeks of age. The animals were divided into two groups: I group (control)--18 rats fed on a standard diet and given tap water ad libitum, and group II (experimental)--42 rats, whose mothers received sodium salt of 2.4 dichlorophenoxyacetic acid in drinking water at a daily dose of c. 250 mg/kg for 2 months before fertilization and during pregnancy and lactation. The animals were killed after 24 hours, 4, 6 and 10 weeks of the experiment. The sections were taken from the kidneys, fixed in 4% formaldehyde and stained with hematoxylin and eosin. For acid and alkaline phosphatase examination, the kidney section were fixed in Backer's liquid and Gomori histoenzymatic reaction was performed. Histological examination of the first four experimental groups revealed changes in kidney tubules. Histologic changes were nonspecific and a variety of conditions. The presence vacuoles in cytoplasm and necrosis of tubular epithelial cells. Varying degrees of isometric vacuolization of proximal tubular epithelium, tubular microfocal calcification, tubular epithelial inclusion bodies and peritubular capillary congestion were observed. The observations suggest that chronic intoxication with 2,4-D acid leads to renal cell damage in kidneys more intensified in the fetal than in the postnatal period. Following herbicide withdrawal, the most pronounced changes observed in the fetus were found to regress. PMID- 12533959 TI - Early detection of hepatic encephalopathy by recording visual evoked potential (VEP). AB - The visual evoked potential (VEP) record in response to a pattern stimulus is a non invasive and reliable method of detecting central and peripheral nerve system abnormalities. VEP recording have been used in animals with fulminant hepatic failure, and also in-patients with hepatic encephalopathy and acute severe hepatitis. Our aims were: a. to evaluate the potency of PVEP in assessing hepatic encephalopathy. b. to find the rate of pathologic PVEP in patients with advanced liver cirrhosis. VEP was recorded in 14 chronic liver cirrhotic patients (6 alcoholic, 6 HCV-related, 2 cryptogenic) and 14 controls. Patients with any neurologic abnormalities were excluded from the study. All patients were subjected to the Mental State Score (MSS) test, and venous blood ammonia was measured on the same day of VEP recording. In 10/14 (71%) patients some VEP recording abnormality was detected. In the cirrhotic patients, P100 latency was significantly longer (P < 0.05) than in controls. Low amplitude was observed in 8 patients compared to controls. Marked increase of N75 (3 patients) and marked increase of N145 (2 patients) were observed. Mean blood ammonia and MSS score were normal in all patients. No correlation was found between both MSS score and blood ammonia levels and the P100 delay. Five out of 10 patients with pathologic VEP developed hepatic encephalpathy during a follow-up of one year, compared to one out of 4 patients with no pathology on VEP recording. VEP recording may be a valuable tool in assessing patients with early hepatic encephalopathy and in predicting encephalopathy. PMID- 12533960 TI - Occurrence of keratinophilic fungus Lagenidium humanum Karling in the surface waters of Podlasie. AB - The occurrence of keratinophilic fungus Lagenidium humanum Karling was investigated, using the bait method, in relation to the environmental conditions in the surface waters of the River Bug, Suprasl and Zwierzyniec in Podlasie in the years 1995-2000. The fungus was found to be the most common at eutrophized sites of the River Suprasl and Biala, and snake exuviae was the most frequently colonized bait. It was revealed that the occurrence of Lagenidium humanum in the water reservoirs was mostly affected by the content of biogenes, suspended solids, oxidability and temperature. Its appearance was seasonal, being most common in spring and autumn. PMID- 12533961 TI - Ultrustructural and enzymatic studies of rat liver after acute cadmium exposure. AB - The aim of the study was to examine morphological and enzymatic changes in the rat liver following single i.v. administration of CdCl2 (5 and 10 mumol/kg b.w.). In the rats given 5 and 10 mumol CdCl2/kg b.w., a correlation was found between the dose and the intensity of changes in the liver both in pathomorphological, ultrastructural and enzymatic analyses. Pathomorphological and ultrastructural investigations showed that a single dose of 10 mumol CdCl2/kg b.w. caused vacuolar degeneration, and fatty degeneration in single hepatocytes. Structural changes were accompanied by an increase in the activity of indicatory enzymes of the liver-AST and ALT. At the same time, stimulation and activation of B-K cells were observed. However, a single 5 mumol CdCl2/kg b.w. dose induced only blurring of cell structure in single hepatocytes within the peripheral zone of the lobules, which looked as if they had been filled with pinkish "mush". PMID- 12533962 TI - Composition of lipophylic extracts from Ficaria verna HUDS. flowers. AB - From the flowers of pilewort--Ficaria verna HUDS. fractions of triterpenes and sterols were isolated by using column chromatography method (CC). The constituents were identified by using TLC, GC and GC/MS methods and by comparison of their retention times and mass spectra with the corresponding data of the authentic samples. The presence of these compounds in F. verna has been described for the first time. PMID- 12533963 TI - Evaluation of lymphocyte populations and subpopulations extracted from inflamed periodontal tissues. AB - The aim of the present study was evaluation the distribution of B- and T-cells and T-cell subsets in periodontal tissues from patients with different periodontitis forms. Periodontal tissue samples were collected from group P patients during routine surgical procedures, while from group C during tooth extraction for orthodontic or prosthetic purposes. Directly after collection, tissue samples were placed in a criostat or fixed in 10% buffered formalin for 24 h at room temperature. Following fixation the material was embedded in paraffin and subjected to routine histological techniques. Examinations of B- and T lymphocytes populations and T-lymphocytes subsets were made with the use of immunohistochemical method. In C group single T and B lymphocytes were found in histological examination in pocket epithelium zones. In early onset periodontitis (EOP) patients in inflammatory infiltration lymphocytes T were dominating while in adult periodontitis (AP) patients dominating were B lymphocytes. Mean CD4/CD8 ratio in control group was 1.7 and in EOP and AP patients 1.1 and 2.6 respectively. PMID- 12533964 TI - Accumulation and loss of Lindane in the muscles, liver and brain of some freshwater fish after intoxication with gamacarbatox suspension. AB - Experiments were conducted under laboratory conditions on young specimens of Carassius carassius L., Cyprinus carpio L. and on adult specimens of Leucaspius delineatus Haeckel. The fish were subjected to a 30-day exposure to gamacarbatox suspension in a dose of 0.5 mg 1-1, containing 10% of lindane as active substance. Directly after the experiment on day 30, and then 40 and 50, lindane content was assessed in the muscles, liver and brain using the method of gas chromatography with an electron capture detector. In all the fish examined, the highest accumulation of lindane and its longest duration were observed in the brain. Less lindane was noted in the liver. The lowest level of lindane was found in the muscles and its concentration in that tissue decreased most rapidly. PMID- 12533965 TI - Effect of maternal smoking on some markers of iron status in umbilical cord blood. AB - Iron deficiency relatively often observed in pregnant women is assumed to be enhanced by cigarette smoking. The present studies are designed to determine the effect of maternal cigarette smoking during pregnancy on iron status of newborns. The levels of ferritin, as well as other iron-markers were determined in placenta tissue and in serum of umbilical cord blood. Placenta tissues and umbilical cord blood from healthy women (n = 30) were divided into smoking and non-smoking groups according to mothers plasma and urine cotinine levels. It is shown that total iron concentration in serum of umbilical cord was similar in both studied groups. In smoking group it was accompanied by higher total iron binding capacity which indicated that functional iron deficiency is possible. Iron storage ferritin in umbilical cord blood was 94 ng/ml and 163 ng/ml in smoking and non smoking respectively. In placenta tissue mean level of ferritin was 252 micrograms/g in the smoking women whereas in the group of tobacco abstinent it was 320 micrograms/g. Low concentration of ferritin both in placenta and umbilical cord blood indicated that smoking during pregnancy could lead to subclinical iron deficiency in matched maternal-cord pairs. PMID- 12533966 TI - The electrophoretical activity of the buccal epithelium cells nuclei in newborns of smoking mothers. AB - The electrophoretical activity of nuclei (EMN) occurs in all cells of the human body. The EMN is variable and depends on the particular stage of human ontogenesis. The factors that modify the ontogenesis may influence the electrophoretical activity of nuclei. The aim of the study was to compare the index of electrophoretical activity of nuclei (index EMN) derived from the buccal epithelium of 40-week newborns (n = 26) from healthy, single pregnancies of smoking mothers with newborns (n = 31) of non-smoking mothers. The cells of the buccal epithelium were analysed using light microscope in the environment of alternating field (voltage 20-30 V; intensity 0.1 mA; frequency 1-2 Hz) generated by Biotest processor. In each group we estimated the number of cells with mobile nuclei and cells with motionless nuclei per 100 cells being observed. The result of this research was that there is not activity of nuclei in the group of newborns of smoking mothers (index EMN = 0 with p < 0.05). The absence of activity of nuclei makes it impossible to use the EMN method for estimation of biological age in the group of newborns of smoking mothers. PMID- 12533967 TI - Correlation between colorectal cancer Bcl-2 expression and tumour clinicopathological variables. AB - The aim of our study was to elucidate whether and in a what way applying two slightly different immunohistochemical procedures effects Bcl-2 protein detection as well as its relationship to colorectal cancer clinicopathological features. A series of 74 primary colorectal cancers was investigated for expression of the Bcl-2 protein. Two immunohistochemical procedures: first with antigen retrieval using a microwave (art-), second with antigen retrieval by use pressure pot (art+) pre-treatment, were applied in each cancer case. Formalin-fixed, paraffin embedded tissues were treated with ant-Bcl-2 antibody (Dako No M 0887). Bcl-2 (art+) immunostaining was positive in 48 (64.9%) cases of colorectal cancers whereas Bcl-2 (art-) immunohistochemical reaction was positive only in 15 (20%) colorectal tumours. A significant, positive correlation was observed between Bcl 2 (art+) expression and localization of the tumour in the rectum (p = 0.007). At the same time a statistically significant positive correlation between tumour Bcl 2 (art-) expression and colon tumour pT stage (p = 0.007), rectum tumour differentiation grade (p = 0.04) and Bcl-2 (art-) protein immunostaining in lymph node metastases (p < 0.000001) was observed. This data suggest that the type of procedure for antigen retrieval to detection of Bcl-2 protein by immunohistochemical reaction, changes the study results and should be considered when comparing own results with the literature data. PMID- 12533968 TI - Bcl-2 immunohistochemical detection in prostate cancer. AB - Prostate cancer (Pca) is the most prevalent cancer diagnosed and the second leading cause of cancer-related deaths among men in the Poland. The present study was designed to analyse the relationship between expression of Bcl-2 protein in prostate cancer (PCa) following radical prostatectomy to chosen anatomoclinical and morfological parameters of the tumours. Tumours from 28 patients were assessed by immunohistochemistry. Tissue sections were fixed in 10% buffered formaldehyde solution, embedded in paraffin and stained immunohistochemically with the anti-human Bcl-2 antibody (Dako/Clon124). The immunolocalization of Bcl 2 was performed using the Labelled Streptavidin Biotin (LSAB) method. No correlation was found between Bcl-2 protein expression and pT stage, lymph node metastases, Gleason score, seminal vesicles invasion, positive or negative resection margins as well as capsular penetration and preoperative PSA serum level. PMID- 12533969 TI - Expression of Ki-67 and PCNA as proliferating markers in prostate cancer. AB - Prostate carcinoma (PCa) is the most common male cancer in industrialized societies and represents a serious public health problem. The aim of our study was the immunohistochemical evaluation of Ki-67 and proliferating cell nuclear antigen (PCNA) expression in prostate cancer (PCa) following radical prostatectomy and analysis of its relationship to chosen anatomo-clinical and morfological parameters of the tumours. Archival sections from 28 PCas were immunostained with monoclonal antibodies against Ki-67, and proliferating cell nuclear antigen (PCNA). Immunolocalization of Ki-67 and PCNA was performed using LSAB method. No statistically significant correlation was found between the expression of Ki-67, PCNA and preoperative PSA level, lymph node metastases, capsular penetration, seminal vesicle invasive and positive or negative surgical resection margins. However, a strong statistically significant correlation between Ki-67 positive and T stage was found. We also found relationship between Gleason score 7 or above and high expression of Ki-67 and PCNA in prostate cancers (p < 0.004, p < 0.02 respectively). These results suggest that PCNA and especially Ki-67 may be useful as tumour markers in prostate carcinoma, but further studies on larger group are required. PMID- 12533970 TI - Comparison of five commercial kits for DNA extraction from human blood, saliva and muscle samples. AB - DNA can be isolated from a variety of human sample sources including anti coagulated whole blood, bloodstains, hairs, tissue samples and buccal epithelial cells. The purpose of the study was to compare yield and quality of DNA samples obtained with the use of five commercial DNA extraction kits. PMID- 12533972 TI - DNA-based paternity testing in Department of Forensic Medicine, Medical Academy of Bialystok, Poland. AB - Results of 255 disputable paternity cases analysed with the use of DNA techniques were reviewed. Probability calculations based on population data has been generated to demonstrate the usefulness of STR multiplex systems for forensic analysis in paternity determination. PMID- 12533971 TI - Serum concentration of TNF-alpha and IL-1 beta in patients with chronic hepatitis C treated with interferon alpha--a preliminary report. AB - The aim of the study was to evaluate the concentration of TNF-alpha and IL-1 beta in 14 chronic hepatitis C patients during interferon treatment in a correlation with biochemical indicators of hepatic cell damage, AspAT and ALAT activity. The study included the sera obtained every month (10-12 measurements) during 48-week therapy. IL-1 beta and TNF-alpha concentrations were determined by ELISA method. Statistic analysis were performed with AnStat program. The wide range of IL-1 beta and TNF-alpha concentration values may prove a differentiated response to immunomodulating treatment with INF-alpha. Concentrations of TNF-alpha, IL-1 beta, show decreasing tendency during therapy. A positive correlation between proinflammatory cytokines (IL-1 beta and TNF-alpha), ALAT and AspAT indicates the role of these cytokines and their relation to a necrotic-inflammatory process demonstrated by aminotransferases concentration. PMID- 12533973 TI - Cardiac groups revise angina management guidelines. PMID- 12533974 TI - One pharmacy's approach to managing drug shortages. PMID- 12533975 TI - On the moon by the end of the decade. PMID- 12533976 TI - AJHP turns 60. PMID- 12533977 TI - White paper on pharmacy technicians 2002: needed changes can no longer wait. PMID- 12533978 TI - ASHP national survey of pharmacy practice in hospital settings: dispensing and administration--2002. AB - Results of the 2002 ASHP national survey of pharmacy practice in hospital settings that pertain to dispensing and administration are presented. A stratified random sample of pharmacy directors at 1101 general and children's medical-surgical hospitals in the United States were surveyed by mail. SMG Marketing Group, Inc., supplied data on hospital characteristics; the survey sample was drawn from SMG's hospital database. The response rate was 46.7%. During 2002, both inpatient and outpatient hours of service increased compared with 2001. Paradoxically, there was an 8.5% decrease in pharmacy staffing and a 7% vacancy rate, suggesting that pharmacists are busier. Most hospitals (80%) had a centralized inpatient dispensing system, but 44% were planning to become more decentralized. Automated dispensing cabinets were used by 58% of hospitals with decentralized drug distribution systems. Most hospitals (81.4%) dispensed more than three quarters of oral doses as unit doses and 63.3% of injectable doses to non-critical care patients, increases from 1999. A large percentage of hospitals (89%) repackaged both oral and injectable medications. More hospitals were repackaging medications than three years ago, primarily because of lack of commercial availability. Approximately 20% of pharmacies either partially or completely outsourced drug preparation activities. Nurses administered medications in virtually all hospitals (99.7%). Despite widespread recommendations to use bar-code technology to check and document doses administered, only 1.5% of hospitals used this technology, an increase from 1.1% in 1999. Nearly two thirds of hospitals used computer-generated medication administration records. While pharmaceutical services are expanding, workforce issues continue to challenge pharmacists trying to maintain and enhance safe medication systems. Safe systems continue to be in place in most hospitals, but the adoption of new technology to improve safety is slow. PMID- 12533979 TI - Characterization of an extemporaneous liquid formulation of lisinopril. AB - The stability of lisinopril in an extemporaneously prepared suspension stored at or below 25 degrees C for 28 days under ambient light exposure was studied. A formulation of 1-mg/mL oral suspension was prepared from commercially available 20-mg lisinopril tablets, using Bicitra and Ora-Sweet SF as the compounding vehicles to make a final volume of 200 mL. Individual samples, stored in 8-oz amber polyethylene terephthalate bottles, were used for each test performed. All samples were stored at 25 degrees C. Appropriateness of the extemporaneous preparation method was performed by shaking three lots of each suspension for 30, 60, and 90 seconds. To test the robustness and reproducibility of the method, two chemists prepared the suspensions from the same three lots of lisinopril tablets. Chemical and physical stability were established by analyzing duplicate samples at time zero and after one, two, four, and six weeks. The solubility of lisinopril was tested from suspensions stored for four weeks. In-use stability was also examined over four weeks. Photochemical stability was examined by exposing three batches of the suspension to maximum light stress in accordance with the International Conference on Harmonization. Antimicrobial-effectiveness testing was also conducted with freshly prepared suspensions and suspensions stored for six weeks. The preparation method used was appropriate and effective. Lisinopril is fully dissolved in the suspension matrix. Satisfactory chemical, physical, and microbiological results were obtained after the suspensions were stored for six weeks at 25 degrees C and 35% relative humidity. Lisinopril suspensions extemporaneously prepared from tablets are stable for at least four weeks when stored at or below 25 degrees C under ambient light exposure. PMID- 12533980 TI - Stability of tiagabine in two oral liquid vehicles. AB - The stability of tiagabine hydrochloride in two extemporaneously prepared oral suspensions stored at 4 and 25 degrees C for three months was studied. Tiagabine is used for adjunctive therapy for the treatment of refractory partial seizures. It is currently available in a tablet dosage form, which cannot be used in young children who are unable to swallow and given doses in milligrams per kilogram of body weight. No stability data are available for tiagabine in any liquid dosage form. Five bottles contained tiagabine 1 mg/mL in 1% methylcellulose:Simple Syrup, NF (1:6), and another five bottles had tiagabine 1 mg/mL in Ora-Plus:Ora Sweet (1:1). Three samples were collected from each bottle at 0, 7, 14, 28, 42, 56, 70, and 91 days and analyzed by a stability-indicating high-performance liquid chromatographic method (n = 15). At 4 degrees C, the mean concentration of tiagabine exceeded 95% of the original concentration for 91 days in both formulations. At 25 degrees C, the mean concentration of tiagabine exceeded 90% of the original concentration for 70 days in Ora-Plus:Ora-Sweet formulation and for 42 days in 1% methylcellulose:syrup formulation. No changes in pH or physical appearance were seen during this period. The stability data for two formulations would provide flexibility for compounding tiagabine. Tiagabine hydrochloride 1 mg/,mL in extemporaneously prepared liquid dosage forms and stored in plastic bottles remained stable for up to three months at 4 degrees C and six weeks at 25 degrees C. PMID- 12533981 TI - Evaluation of compensation of nonphysician providers. PMID- 12533982 TI - Compatibility of fenoldopam mesylate with other drugs during simulated Y-site administration. PMID- 12533983 TI - Patient and parent preference for growth hormone products. PMID- 12533984 TI - Policies on the use of dietary supplements in teaching hospitals. PMID- 12533985 TI - Direct-to-consumer advertising of controlled substances. PMID- 12533986 TI - Safeguards for imaging-related drugs. PMID- 12533987 TI - [Decreased activity in the aging man. When does testosterone help him?]. PMID- 12533988 TI - [Interview with Prof. E. Nieschlag, Munster. How generously do you administer hormones?]. PMID- 12533989 TI - [Prostate carcinoma. Immediate surgery in every case? (interview by Waltraut Paukstadt)]. PMID- 12533990 TI - [Acute coronary syndrome. 4 out of 5 patients are diabetic!]. PMID- 12533991 TI - [Phytogenic drugs in chronic liver diseases. Can mother nature really help here?]. PMID- 12533992 TI - [Neurocardiogenic syncope. Determining the diagnosis with the tilt table test]. PMID- 12533993 TI - [How extensively should irritable bowel syndrome be investigated? Expensive bowel sounds in the abdomen]. AB - Its high prevalence, chronic course, sometimes very troubling symptoms and rather limited therapeutic options, make the irritable bowel syndrome (IBS) one of the most common conditions seen by the family doctor, internist and gastroenterologist. IBS is characterized by abdominal pain accompanied by altered bowel movements and a facultative association with abdominal distension. IBS runs a chronic recurrent course punctuated by relatively symptom-free phases, and an overall good prognosis for survival (quoad vitam). Visceral hypersensitivity, altered gastrointestinal motility, psychosocial factors, and neurotransmitter imbalance are under discussion as underlying pathophysiology. An orienting medical history, clinical examination and exclusion of such alarm symptoms as weight loss, general debility and rectal bleeding, must be followed by a rational choice of further diagnostic measures. PMID- 12533994 TI - [Proper patient counseling, recommended nutrition, specific medication. The basics of irritable bowel syndrome therapy]. AB - In the treatment of the irritable bowel syndrome, it is important to qualify unrealistic expectations with regard to treatment, at an early stage. The therapeutic spectrum encompasses establishment of good rapport between physician and patient, modification of life style, provision of good patient information, reassurance, coping strategies, and temporal restraints on medication. Depending on the leading symptoms, the latter may range from laxatives to probiotics, anticholinergics or spasmolytics, prokinetic and anti-diarrheal agents, to 5 HT3/HT4 receptor antagonists. In individual patients with frequently recurrent or permanent pain, the use of tricyclic antidepressants may be considered. Painkillers should be reserved for patients in whom other therapeutic strategies have failed. PMID- 12533995 TI - [Acute epiglottitis or so-called pseudocroup? Dyspnea is always a true emergency]. PMID- 12533996 TI - [Study of nitrofen as a health risk. Toxic effect on human DNA has been established]. PMID- 12533997 TI - [Left ventricular hypertrophy. Recent aspects of diagnosis and therapy]. PMID- 12533998 TI - [Dermatologic signs of internal diseases. 3: Hyperpigmentation and vasculitis syndrome]. PMID- 12533999 TI - [Presyncope in a 23-year-old women. What was missed in childhood? Atrial septal defect (ASD III)]. PMID- 12534000 TI - [Efficacy trial in general practice substantiated. New antibiotic with strong effectiveness and tolerance]. PMID- 12534001 TI - [Osteoporosis therapy in general practice. Which measure should be included?]. PMID- 12534002 TI - [Calcium antagonist against hypertension. Arteriosclerosis controlled]. PMID- 12534003 TI - [Breast carcinoma. Preoperative chemotherapy can preserve the breast]. PMID- 12534004 TI - [Young patient with stress dyspnea of uncertain origin. Evaluate for pulmonary hypertension!]. PMID- 12534005 TI - [Respiratory tract infections. Increasingly more established antibiotics fail]. PMID- 12534006 TI - [Coronary heart disease prevention in type 2 diabetic patients. What is the value of fibrates?]. PMID- 12534007 TI - [The human is becoming increasing more transparent. Tumor-free intestine thanks to color images]. PMID- 12534008 TI - [Increased transaminases but usual hepatitis diagnosis is negative. Immune system as liver killer?]. PMID- 12534009 TI - [Anti-aging medicine and cosmetic surgery. Complications are too often not mentioned]. PMID- 12534010 TI - [Minor dyspnea with exertion--but extremely poor prognosis. What reveals pulmonary hypertension?]. PMID- 12534011 TI - [Suggestion--salicylic acid--travel. What really helps control warts? (interview by Dr. Thomas Meissner)]. PMID- 12534012 TI - [Patients with symptoms of dementia. Consider more than the brain!]. PMID- 12534013 TI - [Hypotension--harmless or dangerous? The "orthostasis test" gives the answer]. AB - Patients with non-specific symptoms that manifest when standing, are commonly seen in the doctor's office. The suspected diagnosis orthostatic dysregulation, can be established on the basis of a careful history, physical examination and a standing test. Frequent symptoms of orthostatic dysregulation are orthostatic hypotension, neurocardiogenic syncope and orthostatic intolerance. If a disorder of the autonomic nervous system is suspected, the need for further investigations is evaluated on an individual basis. The aim of such further diagnostic procedures is to differentiate between mild forms of autonomic disturbance that are unlikely to affect the patient's prognosis (mild orthostatic intolerance, neurocardiogenic syncope) and severe forms of autonomic failure, to identify the affected parts of the autonomic nervous system (partial or global autonomic failure) and, whenever necessary, to initiate appropriate treatment. PMID- 12534014 TI - [Effective "home remedies" in orthostatic hypotension. A glass of water before getting up and a brick under the bed]. AB - The symptoms of orthostatic hypotension can be considerably improved by non pharmacological treatment, and patients with such symptoms should not initially be given medication. Many patients, for example, show a pronounced increase in blood pressure shortly after drinking water, and this phenomenon can be utilized to ameliorate symptoms. In patients who respond inadequately, treatment with pharmacological agents is indicated. The most appropriate drug needs to be selected individually on the basis of its action and tolerability. All medications suitable for treating orthostatic hypotension can appreciably increase the blood pressure in the prone patient. PMID- 12534015 TI - [When blood pressure regulation seems to make no sense. Think of baroreceptor reflex failure!]. PMID- 12534016 TI - [Rectal hemorrhage. How much blood has your patient already lost?]. PMID- 12534017 TI - [71-year-old patient with subacute cough and middle lobe atelectasis. A hard nut with benign clinical course]. PMID- 12534018 TI - [Often inadvertently burned. Phototoxic reaction of the skin]. PMID- 12534019 TI - [Swelling of the head area after intubation. What went wrong? Tracheal rupture]. PMID- 12534020 TI - [A frequently underestimated dermatomycosis. Long-term therapeutic success with fungicidal effect]. PMID- 12534021 TI - [Fungi colonizing heads of children. Treating tinea capitis properly]. PMID- 12534022 TI - [New onset headache. Which patients should be sent for CT imaging?]. PMID- 12534023 TI - [Obesity and hypertension. A dangerous liaison]. PMID- 12534024 TI - [Ventricular arrhythmia. A step-wise treatment plan]. PMID- 12534025 TI - [Interpretation of hypertransaminasemia]. AB - Raised serum level of transaminases aspartate (aminotransferase and alanine aminotransferase) is a frequent situation in medical practice. It is considered as moderate when the level is under 10 times normal and as chronic when it lasts for more than 6 months. The most common etiologies for chronically elevated transaminases are alcohol use, viral hepatitis, liver steatosis, diabetes, obesity and medications. Many non invasive tests, including history, physical examination, blood tests (markers for hepatitis A and B, muscular enzymes), and imaging procedures (abdominal ultrasonography) are usually done and lead to a correct diagnosis in 80% of patients. When the diagnosis cannot be determined non invasively a liver biopsy is recommended in order to make diagnosis, to evaluate the prognosis and to start an adapted treatment. PMID- 12534026 TI - [Epidemiology of gram negative bacterial septicemias: data from a Tunisian hospital (1996-1998)]. AB - Gram-negative bacilli (GNB) septicemia are among the most serious infections encountered in the hospital since they generally occur on debilitated patients and are due to the multi-drug resistant bacteria. A retrospective study relating to 195 septicemia was carried out with an aim studying epidemiologic profile, predisposing factors, entry sites for micro-organisms, responsible GNB and their antibiotic susceptibility. GNB septicemia were mainly frequent in intensive care units (34%) and surgery (31%). Previous antibiotherapy, invasive procedures and surgical acts were the principal predisposing factors. The entry sites for micro organisms remained unknown in 1/3 of the cases. The most common source of septicemia was the urinary tract infections. E. coli was the most frequent isolated bacteria (26%) in the community acquired spticemia whereas Klebsiella Enterobacter-Serratia (KES), Acinetobacter and Pseudomonas were mainly encountered in nosocomial infections. Imipenem remained the most active betalactamin on GNB (2% of resistance) with amikacin (16% of resistance) among aminoglycosides. The rate of mortality was 18%. Hospitalization wards (intensive care units, surgery), entry sites unknown, septic shock syndrome were the main factors of prognosis. The development of immunology and molecular biology should improve the outcome of these infections but the preventive measures remain the most effective. PMID- 12534027 TI - [Attitudes and practices in the Sahelian Tunisian population regarding drug usage]. AB - Drugs occupies a strategic place in health care expenses. However, irrational practices characterize its utilization by all actors and particularly the population. The objective of this populational study is to identify attitudes and practices of the households of the sanitary region of Sousse (Tunisia) in the domain of medicine use. It is a descriptive survey concerning a representative sample of 402 households distributed in four localities (two urban, one semi urban and one rural). Data have been collected through a questionnaire managed during an interview with the family's member (most often the chief of the household). We found that beliefs of the population are often erroneous (the expensive medicines are considered more efficient by 52% of interrogated persons and practices were often maladjusted (Self medication, insufficient compliance). A community mobilization via an education of health oriented by results of this exploratory survey would be an important component of the global strategy of promotion of the rational use of drugs in our country. PMID- 12534028 TI - [Anti-erythrocyte alloimmunization in an obstetrical milieu. Study of 2093 cases]. AB - From January 94 to December 99, we were looking for an anti-erythrocytary immunization at 2093 women followed up in obstetrical environment. As a whole, 2232 researches of antierythrocytary antibodies (RAI) were carried out, say 1.07 RAI/women. These RAI concerned, in 4 cases out of 5, some Rh- women having an Rh+ spouse. The results were positive among 78 women, say 3.73% and 71 women having an allo-immunization antibodies (Ab), say 3.39%. The specificity of the Ab is variable: 45 anti-D, 33 Ab other specialities (C, E, K, Fya, JKb, Lua,...) and 16 positive RAI which are difficult to interpret. Besides, the immunization of women according to the number of gestations shows an increasing rate ranging from 0.27%, for a gestation to 7.44% for 4 gestations or more. Finally, the prophylaxis of the allo-immunization to the antigen D, possible thanks to the immunoglobulins anti-D, was only active among 42.13% of Rh women having a Rh+ spouse. PMID- 12534029 TI - [Value of prenatal corticotherapy in the prevention of hyaline membrane disease in premature infants. Randomized prospective study]. AB - OBJECTIVE: The aim of the study was to determine the feasibility, the cost and the effects of antenatal maternal corticosteroid treatment on preventing respiratory distress syndrome in premature neonates of our population. SUBJECTS AND METHODS: Between January, 1, 1998 and June, 31, 1999, 118 pregnant women at 26-34 weeks' gestation and at a high risk of premature delivery, were prospectively randomized in 2 groups: group 1 received intramusculary 24 mg of betamethasone (12 mg every 24 hours), group 2 didn't receive antenatal corticosteroids. At birth, premature neonates were systematically examined by a neonatologist. RESULTS: 131 premature neonates were born (63 from group 1, 68 from group 2). The incidence and the degree of severity of respiratory distress syndrome, appeared substancially reduced (4.8% vs 27.9%) by the use of antenatal corticosteroids. Moreover, neonatal mortality due to respiratory distress syndrome was statistically less in group 1 than in group 2 (22.9% vs 57%). There was no significant difference in the occurrence of maternal or neonatal corticosteroid complications such as infection between treated group and control subjects. We estimated a potential annual savings of 21 thousands tunisian dinars, when the cost implications for antenatal corticosteroid therapy were estimated to 2 thousands tunisian dinars. CONCLUSION: Maternal administration of corticosteroids before preterm delivery results in a decrease in the incidence and severity of respiratory distress syndrome and a decrease in neonatal mortality rate among premature neonates born to treated versus untreated mothers at 26-34 weeks' gestation; added to an annual savings estimated to 21 thousands tunisian dinars. PMID- 12534030 TI - [Erosive buccal lichen planus: nine case reports]. AB - Erosive oral lichen planus (EOLP) is uncommon. We present nine cases of EOLP in patients aged from 39 to 77 years. The aim of our retrospective study was to determine the clinical particularities and the treatment difficulties of this disease. PMID- 12534031 TI - [Diagnostic and surgical strategy for parathyroid adenoma]. AB - From a retrospective study carrying on 8 patients operated for sporadic parathyroid adenoma, we underline the place of medical imagery which allowed the detection of parathyroid adenoma in 7 cases. For all our patients the exploration of thyroid space was bilateral, seeking parathyroid glands. The authors discuss the stages and the various processes of the surgical strategy for parathyroid glands' exploration. PMID- 12534032 TI - [Prognostic evaluation of histological criteria in central nervous system astrocytomas: 51 cases]. PMID- 12534033 TI - [Eclampsia perinatal mortality at the Tokoin University Hospital Center (Lome Togo): report on 91 newborns]. AB - A one-year (January 1-december 31, 1998) case control study was carried out at the Gynaecology and Obstetrics ward and at the Paediatrics ward of the Lome Tokoin Teaching Hospital (Togo). Through following parameters (delivery, sex, pregnancy age at delivery, birth weight, early neonate period issue, moment and number of fits) newborns from 89 mothers with eclampsia were compared to those from 445 pregnant women without high blood pressure admitted during the study period. After every woman with eclampsia (study group) the five following without high blood pressure (control group) were included in the study. Relative risk (95% confidence interval) and chi 2-test (5% significant degree) were calculated in the comparison of some parameters. RESULTS: The perinatal mortality rate was 28.6% (26 out of 91 newborns) in the study group versus 8.8% (44 out of 455) in the control group (relative risk = 2.91; chi 2 = 27.87; p = 0.0001). Birth weight, pregnancy age at delivery and fits number before delivery influenced significantly perinatal mortality rate; meanwhile the lower was the relative risk of eclamptic fits onset, the more frequent were ante-natal visits. CONCLUSION: The perinatal mortality rate due to eclamptic might be reduced through frequent ante-natal visits, sustained ante- and post-natal care specially adequate management of eclamptic patients and their newborns. PMID- 12534034 TI - [Prevalence of anti-GAD autoantibodies in Tunisian children with type 1 diabetes]. AB - The prevalence of GAD antibodies and its correlation with some autoimmune markers: ICA (islet cells antibodies), antinuclear antibodies, thyroid antithymune and antimicrosomes antibodies, were studied in 84 Tunisian type 1 diabetic children. The prevalence of GAD antibodies was 51.2% and decreased as a function of increasing duration of the disease. Their frequency was 84.6% in children with newly diagnosed diabetes (within 6 months of diagnosis) and only 29.41% in those with a longer duration of the diabetes (more than 5 years). ICA were present less frequently (21.4% of the children). 10.7% of the studied samples were positive with the two antibodies (GAD ab ICA), and 40% were positive only with GAD antibodies. We conclude that the GAD antibodies seems to be more associated to the development of type 1 diabetes than ICA. They are detected more frequently in patients with long standing disease, that's make their determination very interesting as diagnostic and predictive marker. PMID- 12534035 TI - [Haemophilus influenzae purulent meningitis in adults: looking for a predisposing factor]. AB - We bring back an adult case of purulent meningitis to Haemophilus influenzae. We insist on the particular aspects of the host of this meningitis type at the adult. These aspects must be searched every time that Haemophilus influenzae is isolated in cerebrospinal fluid in adult's meningitis. PMID- 12534036 TI - [Fatal postoperative crisis in acute hereditary porphyria]. AB - The acute porphyria is an autosomal dominant disorder of the heme biosynthesis enzyme. The aim of this work is to determin a diagnostic step in order to anticipate porphyric crises. We report the observation of a child which was hospitalized in the pediatric service three years ago for peripheric arthralgies and myalgies where the diagnosis of an acute porphyria was not posed. He wase admitted in our service for appendicitis, he was operated, the anesthesie was carried out by the thiopenthal and the succinylcholine. At the first post operative day, a respiratory insufficiency was the cause of death. The rate of coproporphyrines and of uroporphyrines in the urine had confirmed the diagnosis of acute porphyria. The positive diagnostic of acute porphyria is difficult, the association of abdominal, psychological and neurologic signs must suggest the diagnostic, all the more, if the urines have a dark colour. The presence of uroporphyrines and coproporphyrines and certain precursors in the urines or in the high-rated stools confirm the diagnostic. The family survey and dosage of porphyric compounds in the sickness-bearing patient's family members allow to reveal certain asymptomatic forms of the heriditary acute porphyria, it is caused by certain products used in neuropsychiatry, in aneasthesic or in certain circunstances, such as, the infection or the trauma. The treatment of the acute porphyria is an emergency wich requires the transfert of the patient to a unit of intensive care. PMID- 12534037 TI - [Splenic angioma. Value of MRI: two case reports]. AB - The splenic angioma is a rare tumour. Its aspects MRI are still badly known. The authors show that in the 2 brought back cases, there is a hypersignal on the 3 echoes of T2 with a catch of contrast slow and centripetal after Gadolinium injection. The hemorrhagic risk in connection with a spontaneous or traumatic rupture imposes the identification of this lesion. PMID- 12534039 TI - Heparin use during pregnancy. AB - The use of anticoagulants in pregnancy is problematic. In this focus, the authors reviewed the problems associated with the use of vitamin K antagonist during pregnancy. Then, they presented the general considerations on the use of unfractioned heparin and low molecular weight heparin during pregnancy. Finally they established the therapeutic recommendations for anticoagulation during pregnancy including treatment and prophylaxis of venous thromboembolism, anticoagulation of pregnant women with mechanical heart valves and the management of pregnant women with antiphospholipid antibodies and multiple pregnancy losses. PMID- 12534038 TI - [Primary renal non-Hodgkin lymphoma]. AB - Primary renal non Hodgkin lymphoma is rare and unusual because the renal parenchyma does not have lymphatics. We report a case of bilateral primary lymphoblastic T lymphoma presenting with renal insufficiency in a 14 year old girl. We discuss clinical, pathological particularity and prognosis. PMID- 12534040 TI - [Transfusion practice and attitudes]. AB - During last years, blood transfusion and transfusion security were domains which have preoccupied health authorities, the population and health professionals. Beginning with the principle that blood transfusion is a care that must be under medical responsibilities, authors analyse physician's implication in this field. To reach this objective, a study interested 199 young physicians, working in two university hospitals in Tunisia. Results indicate that physician's knowledge about blood transfusion is insufficient to guarantee quality of this medical act. In terms of attitudes and practices, physicians are not really always implicated in the different stages of blood transfusion process. Intervention is limited to prescription and blood transfusion record card writing. Transfusion herself and monitoring were, in particular, delegated to nurses. So transfusion incidents were under--declared. Transfusion medicine must occupy important place in physicians curriculum. At hospital level, quality insurance process, based on audit, have to be implemented to obtain transfusion good practices, in a domain were risks are unfortunately not yet totally circumscribed. PMID- 12534041 TI - [Value of cytology in the diagnosis of thyroid nodules (93 cases)]. AB - This is a prospective study of 93 thyroid cytologic specimen from the department of ORL of Charles Nicole Hospital and analysed by the department of Pathology of the same Hospital. The purpose of this study is to evaluate the accuracy of thyroid cytology in the diagnosis of thyroid nodules. The age average in our study was 40 years with a male-female ratio of 1/6. Thyroid cells were collected by a fine needle aspiration. The following results were obtained: an interpretability rate of 7.52%, a sensitivity of cytological examination as compaired with histopathological examination of 70% and a specificity of 97.43%. PMID- 12534043 TI - [Central agenesis of the hand]. AB - Central aplasia represent under 10% of congenital malformations of the hand. It is characterised by the partial or complete absence of the median ray. We adopted the classification of Swanson and considered this anomaly a stop of median longitudinal development. We separate 2 groups: Hand in folk: the aplasia interested only the median digits, all metacarpal are present. Hand in lobster's pincer: all median rays are absent including the metacarpal. We report a set of 16 children carriers of 23 central aplasia of the hand, it was about 14 hands in lobster'pincer and 9 hands in folk. In any case, the gene was merely aesthetic, the hand's function was complete. A child has been operated, and benefitted a closing of the median crack defect, he had a free thumb. For the other cases, abstention has been decided, indeed to close this cracks deprive these hands of a first corner the alone present. PMID- 12534042 TI - [Delayed growth due to growth hormone deficiency (study of 16 cases)]. AB - Growth hormone deficiency is one of the scarce statural backward causes. It is difficult to make a diagnosis. The purpose of growth hormone treatment is to reach a final normal height and to avoid hypoglycemia after-effects. We give a retrospective account of 16 children (11 boys and 5 girls) who have a growth delay due to a total growth hormone deficiency confirmed by the stimulation tests and who have also been given benefit of a biosynthetic growth hormone from 1990 to 1999. The statural backwardness varies from--2.5 DS to--4 DS, with an average of--3.5 DS. In all the cases it is a matter of a harmonious backwardness discovered at an average age of 6 years. The bone age has revealed an important backwardness of bone maturation: average bone age of 3 years for boys and 4 years for girls. The hormone balance sheet reveals, in all the cases, a total growth hormone deficiency (GH < 5 ng/ml) combined with a corticotrope deficiency (2 cases) and thyreotrope (3 cases). It is a growth hormone idiopathic deficiency among our patients. The growth hormone treatment has been administered at the average age of 8 years. The weekly doses were (0.4 to 0.8 U/kg). The evolution was favorable with an average growth speed that has gone up from 3 cm/year before the treatment to 10 cm in the first year of the treatment and to 5.5 cm during the second year of the treatment. An average statural gain of 0.8 DS and a bone maturation gain of one year over one year treatment. The authors put into relief the importance of diagnosis criteria of growth backwardness through a GH deficiency and suggest a therapeutic diagram, and a follow-up of the GH biosynthetic treatment. PMID- 12534045 TI - [Subretinal fluid drainage in rhegmatogenous retinal detachment]. AB - Subretinal fluid drainage during retinal surgery is a tricky manoeuver that remains controversial. In this study, we carried out a comparative retrospective study about 265 cases with retinal detachment that had a primary episcleral surgery. We compared the results of the group operated with subretinal fluid drainage (199 cases) to the group without (66 cases). The complications met were retinal haemorrhages in 14 cases (7%) and retinal incarceration in one case (0.5%). Anatomical results were 91.45% in the group with drainage and 93.93% in the group without. There was no statistical difference between the two groups. The complications of the subretinal fluid drainage are extremely rare and have no repercussion if it is well checked. Subretinal fluid drainage remains the surgeon decision. It is achieved by most retinal surgeons (75%). PMID- 12534044 TI - [Epidemiology of osteomyelitis and arthritis in infants]. AB - A retrospective study realised over 10 years in the orthopaedic unit of the children's hospital of Tunis allowed to collect 206 cases of osteomyelitis and arthritis in infant (3 months--3 years old). In 74.6% of the cases, infection involved the joint. There is a single focus of the infection in 92.6% of the cases, more often at the lower limbs. The pathogen has been isolated in 33% of the cases. Staphylococcus aureus (52 %) is the more frequent pathogen, especially in osteomyelitis. Enterobateria (16%), Streptococcus (17.7%) (Streptococcus pneumonia: 10.7%), and Hemophylus influenzea (5.3%) were often isolated in arthritis. PMID- 12534046 TI - [Value of reagent strips in screening urinary infection in children]. AB - The aim of this study is to specify the role of rapid tests in the screening of childhood urinary tract infection. During the period between july to december 1998, 572 urinary samples were collected from pediatric out-patient in Hopital d'Enfants de Tunis and aged from 1 month to 15 years. Only 75 samples (12.5%) were culture positive. The predictive value of leucocytes or nitrites test was 97.2%. These results allowed the use of rapid test in the screening of urinary tract infection in children. However, if clinical symptoms are present, the culture of urine must be associated to the rapid test. PMID- 12534047 TI - [Retinal detachment due to macular hole: therapeutic problems]. AB - To compare the success rate and the visual improvement of different surgical techniques for treating 21 retinal detachments due to macular hole in highly myopic eyes. METHODS: Twelve patients were treated by gas tamponade alone, 5 patients underwent vitrectomy with gas tamponade, 4 patients underwent vitrectomy with epiretinal membrane pealing and gas or oil silicone tamponade. RESULTS: Fifty eight % of patients were successfully treated after gas tamponade alone, and 75% after vitrectomy, epiretinal membrane pealing and gas or silicone oil tamponade. After retreatment in 7 patients, the final reattachment rate was 80%. Visual acuity was inchanged in 12 cases, improved to 1/20-1/10 in 8 case and reached 3/10 in 1 case. CONCLUSION: A therapeutic scheme is suggested according to the type of retinal detachment, macular vitreoretinal traction and pigment epithelium and chorioretinal degeneration condition. PMID- 12534048 TI - [Epilepsy with affective symptomatology: three case reports]. AB - Disagreeable affect of fear or anxiety can form the essential of the epileptic critical semiology. The centrum seat is most often found to be temporal and in some rare cases frontal. This form of epilepsy makes it sometimes difficult to establish a differential diagnosis specially when confronted with panic attack. A careful semiologic analysis is needful. Some clinical criteria are in favor of an epileptic etiology like the shortness of the crises, the stereotyped characteristic of the semiology or the association with some other epileptic symptoms. The assumption of neurophysiologic substratum common to both entities is put forward by the authors. Indeed, the current researches are moving towards the likelihood that the limbic structures are stimulated by the epileptic discharge. These structures are taking a part in the pathogeny of the anxiety troubles. Three clinical vignettes are illustrating the difficulties in diagnosing this form of epilepsy. PMID- 12534049 TI - [Mitral valve prolapse and sudden death: a case report]. AB - Mitral valve prolapse (MVP) is a common disorder that, in general, has a good prognosis. Rare occasions of sudden death have been reported in patients with MVP and it is presumed that the basis of sudden death is arrhythmias. We report a case of a 47 years old men affected by MVP complicated by ventricular arrhythmias and sinoatrial block; who died suddenly from ventricular tachycardia. The pathophysiology and risk factors of sudden cardiac death in MVP are discussed. PMID- 12534050 TI - [Pityriasis rotunda: second Tunisian case]. AB - Pityriasis Rotunda is a dermatosis well known in Japan and Black Africa. Its unusual in Europe and North Africa. Several authors consider that Pityriasis Rotunda is an acquired ichtyose and that it represents a real paraneoplastic syndrome. We report the case of Tunisian men suffering from a Pityriasis Rotunda associated to a dilatation of branch. PMID- 12534051 TI - [Complete utero-vaginal duplication with hematocolpos and homolateral renal agenesis]. AB - We relate one observation about a rare case of genital malformation, where is on association of unilateral hematocopos, bicorn uterine and homolateral renal agenesis. The diagnostic was suspected upon the association with a progressive dysmenorrhe and vaginal mass. II was confirmed by ethnographie and IRM. The treatment has been conservative and consisted only in the section of the vaginal partition. PMID- 12534052 TI - [For the family physician allergy season is all year long. Strategies against running noses]. PMID- 12534053 TI - [To unmask allergens. Prick or specific IgE?]. PMID- 12534054 TI - [A case for proton pump blocker... When cough comes from the stomach]. PMID- 12534055 TI - [Painful arm lymphedema after breast cancer surgery. Can surgery still help?]. PMID- 12534056 TI - [Promises of Functional Food are disputed. Can one eat to become healthy?]. PMID- 12534057 TI - [Post-discharge surgical wound infection: what is to be done?]. AB - Infection of the laparotomy wound cannot always be prevented--for a variety of reasons including patient-related, disease-related and surgery-related factors. Of importance is the early detection of a wound infection--with confirmation or exclusion of underlying pathology (e.g. anastomotic leakage)--before sepsis develops. As a rule, a laparotomy wound infection can be adequately controlled by re-opening, debridement and moist management. Innovative dressing techniques involving vacuum sealing and suction accelerate healing. PMID- 12534058 TI - [Abdominal pain following laparotomy. When does the patient require adhesiolysis?]. PMID- 12534059 TI - [Postoperative fistulas. How to close them]. AB - Postoperative enterocutaneous fistulas are usually the result of an anastomotic leak. The likelihood of a postoperative fistula developing depends on concomitant diseases. An established gastrointestinal fistula is associated with considerable morbidity and mortality. Appreciable losses of fluid, protein and electrolytes often result in malnutrition, which has a negative impact on wound infection and the mental performance of the patient. Conservative treatment must aim to compensate these deficits as early as possible. Adequate amounts of energy (carbohydrates and fats) vitamins and trace elements must be ensured by replacement measures. In addition to appropriate drainage of the intra-abdominal focus and the use of antibiotics, treatment with octreotide (somatostatin analogues) may be considered. For conservative treatment, however, free intestinal transit distal to the fistula must be ensured. If an enterocutaneous fistula fails to heal within two or three months under conservative treatment, surgical management must be applied. PMID- 12534060 TI - [Is one antihypertensive agent as good as another?]. PMID- 12534061 TI - [Ophthalmological emergencies. 2. Trauma and acute loss of vision]. PMID- 12534062 TI - [Acute abdominal pain. Huge what was brought to light!]. PMID- 12534063 TI - [Classic nitrate PETN (pentaerythritol tetranitrate). Initial data in pulmonary hypertension: no tolerance]. PMID- 12534064 TI - [Atherosclerosis progression and standard in coronary disease. Nitrate with special talents: PETN (pentaerythritol tetranitrate) is vasoprotective]. PMID- 12534065 TI - [When have you thinned out your insurance last time? Don't throw out any Euros through the window!]. PMID- 12534066 TI - [Chronic hepatitis C. A new chance with Consensus Interferon for patients with handicaps]. PMID- 12534067 TI - [Beta blockers and ACE inhibitors In heart failure to be combined at the beginning!]. PMID- 12534068 TI - [Convenient, effective and well accepted. The weekly pill against osteoporosis]. PMID- 12534069 TI - [In acute coronary syndrome. Seize platelets with pliers]. PMID- 12534070 TI - [From harmless to life threatening--increasingly more people suffer from mycoses. Fungi from head to foot]. PMID- 12534071 TI - [Neglected foot mycosis. Erysipelas is a threat!]. PMID- 12534072 TI - [(Co-) etiology of chronic sinusitis. Fungi in the paranasal sinuses]. PMID- 12534073 TI - [From saprophytes to pathogens. Fungi with an appetite for lipids]. PMID- 12534074 TI - [Systemic fungal infections in severely ill patients. When opportunistic pathogens decided between life or death]. PMID- 12534075 TI - [A new drug approach approved in the USA. Cox-2 inhibitor in intestinal cancer]. PMID- 12534076 TI - [WHO intends to eliminate measles--but we fall short. The German vaccination baffle]. PMID- 12534077 TI - [Left or right. Which testicle hangs lower?]. PMID- 12534078 TI - [Health care reform. An idiotic expression!]. PMID- 12534079 TI - [Proton irradiation of tumors spares surrounding tissue. A new era in cancer therapy? (interview by Dr. Judith Neumaier)]. PMID- 12534080 TI - [In many hypertensive patients the systolic pressure remains uncontrolled. Is the selection of 140 mm Hg as the goal value too low?]. AB - Despite the best preconditions, adequate control of arterial hypertension remains a stub born problem in the primary prevention area. Less than optimal control of hypertension is not confined to Germany--rather too low rates of detection and treatment have long been a problem for health care providers. But hypertensive treatment also inadequately succeeds in lowering systolic pressure to below the target level of 140 mmHg. Close study of the data provided by clinical trials also reveals that even intensified treatment, including the use of combinations of drugs and increased doses of antihypertensives, fails to reduce the systolic value to below 140 mmHg, while the elevated diastolic values can be brought down in more than 90% of the cases. These facts suggest that the practical significance of the target systolic value needs to be discussed. The wish to achieve the best possible control should not tempt us to close our eyes to the realistic potentials--and limitations--of medical care. PMID- 12534081 TI - [When blood pressure control is a problem. Refractory hypertension is rare]. AB - Hypertension is considered refractory when the use of three pharmacologically different antihypertensives (included a diuretic, a sympathetic blocker and a vasodilator) given at maximum dose fails to lower the blood pressure to below 140/90 mmHg. In the event of isolated systolic hypertension, the value should be reduced to below 140 mmHg. A truly refractory form may be assumed in some 2-5% of all hypertensives, and must be differentiated from a pseudo form in which such factors as counter-regulatory mechanisms, inadequate use of general measures, inadequate antihypertensive therapy, interactions with other drugs, or secondary hypertension may play a role. Refractory patients in whom the underlying cause cannot be identified should be referred to an appropriate specialist. Diuretic treatment should first be optimized and antihypertensive doses maximized (with consideration being given to side effects). Further substances such as minoxidil in combination with a beta-blocker and a diuretic may be used. PMID- 12534082 TI - [Ideal combination partners in therapy of hypertension. Calcium antagonists are much better than their reputation]. AB - Calcium antagonists are a chemically heterogeneous group of substances that effectively reduce elevated blood pressure in all age groups and also show organoprotective properties. A number of randomized studies have revealed a reduction in morbidity, in particular stroke. These substances are the drugs of first choice in the elderly, in left-ventricular hypertrophy and in obstructive pulmonary disease. Care must be exercised when congestive heart failure or acute coronary syndrome presents. Calcium antagonists are ideal combination drugs that help achieve targeted blood pressure levels. PMID- 12534083 TI - [Emergencies in general practice. Acute hearing loss]. PMID- 12534085 TI - [Pilot study with pyrimidine nucleotides. Less pain in diabetic polyneuropathy]. PMID- 12534084 TI - [Questionable group concept. How interchangeable are so-called analog drug preparations?]. AB - Medical drugs are usually grouped together because they exhibit identical or similar chemical and pharmacological properties. Thus, for example, all known statins lower morbidity and mortality in coronary patients. In the meantime, three beta blockers have been shown to have a life-prolonging effect in mild-to moderate cardiac insufficiency. The question is: to what extent are the preparations interchangeable? The beta blockers in particular manifest numerous other properties--such as beta 1 selectivity, vasodilatory or antioxidative actions which, under certain circumstances may even be decisive for the success or failure of treatment. The mortality-lowering effect of high-dose ACE inhibitors in patients with a high cardiovascular risk has to date, been shown only for a single preparation. For others, the high dose (effect) has yet to be confirmed. Also, with every new substance, a new side effect profile is introduced, which may result from the tiniest alteration to the active agent molecule, but also by a different preparation method. In consequence, unexpected effects, such as allergic reactions, may occur. This not only makes the grouping of similar substances, but also the interchange of "me-too" or generic preparations, problematic. PMID- 12534086 TI - [Rheumatic disease therapy without regret. Meloxicam dosage safety]. PMID- 12534087 TI - [No loss of effectiveness with rofecoxib. Long-term control of rheumatoid pain]. PMID- 12534088 TI - [Improved antilipemic drug is approved. Cholesterol absorption is inhibited]. PMID- 12534089 TI - [These hypertensive patients are especially at risk for myocardial infarct in the morning. Screen for non-dippers]. PMID- 12534090 TI - [Selective calcineurin inhibitor. Neurodermatitis patients need fewer corticoids]. PMID- 12534091 TI - [Thanks to fast acting insulin analogs, diabetes therapy without hourly schedule]. PMID- 12534092 TI - [Chemotherapy improves prognosis in colorectal carcinoma. Soon oral therapy at home?]. PMID- 12534093 TI - [Effective treatment of migraine attack. But how to deal with headache recurrence?]. PMID- 12534094 TI - [Effective NSAID as optically pure isomer. Therapy of arthrosis becomes more tolerable]. PMID- 12534095 TI - [37. Each jaundice incident is an alarm signal!]. PMID- 12534096 TI - [The mammary gland, an endocrine gland]. AB - Features of acromegaly had developed in a 6-year-old crossbred Belgian shepherd dog (dog A) following treatment with medroxyprogesterone and an 8-year-old beagle dog (dog B) during the luteal phase of the oestrus cycle. Discontinuation of progesterone administration in dog A and ovariohysterectomy in both dogs was followed by a regression of the soft tissue changes and a normalisation of the initially high plasma growth hormone concentrations. These cases illustrate that the mammary gland is not just a gland which delivers milk in response to certain stimuli, but that it also transmits humoral signals, i.e., the mammary gland is an endocrine gland. Progestin-induced growth hormone is produced in areas of hyperplastic mammary epithelium and in mammary tumours. This suggests that locally-produced growth hormone is involved in the proliferation of unaffected and tumorous mammary tissue in an autocrine or paracrine manner. Now that the expression of the growth hormone gene has also been demonstrated in the human mammary gland, further studies are needed to answer questions such as (a) does mammary growth hormone also reach the systemic circulation in women?, (b) what is the role of local growth-hormone production in tumourigenesis?, and (c) does this phenomenon provide possibilities for therapeutic intervention in women with breast cancer? PMID- 12534097 TI - [How much alcohol is too much and why? Comments on socially accepted but excessive alcohol drinking patterns]. AB - Many modern societies increasingly seem to accept drinking patterns that should be qualified as excessive from a health care point of view. This does not concern alcoholism or alcohol addiction but a daily intake of three to eight glasses and the pattern of binge drinking. Although a lot of data are available on the physical consequences of alcohol abuse, little is known about the impact of these drinking patterns on peoples' health. According to the J-shaped curve between alcohol consumption and mortality, an increase in morbidity and mortality can be noticed from two to three drinks a day. This is largely caused by an increase in accidents, malignancies, cardiovascular diseases and neurological disorders. However, data on pathophysiological mechanisms and on the contribution of these drinking patterns in medical consumption are scarce. Because of the estimated extent of the problem and societies' interest, more support should be given to scientific programs and research into this matter. This may not only contribute to a better monitoring of the problem but also guide the development of strategies for education and prevention. PMID- 12534098 TI - [Nutrition and health--favorable effect of wine and wine flavonoids on cardiovascular diseases]. AB - Epidemiological studies have shown a favourable effect of moderate alcohol consumption with regard to atherosclerotic disorders. In addition to alcohol, wine contains a large number of other components including polyphenols. These polyphenols mainly originate from the skins and seeds of grapes and, because of differences in vinification, their variety and concentration is higher in red wine than in white wine. In vitro and ex vivo studies have shown that some of these polyphenols are able to slow down LDL-cholesterol oxidation, stimulate NO production, influence prostaglandin synthesis and inhibit platelet aggregation. However, little is known about their resorption, bioavailability and effectiveness in vivo. Since data from intervention studies with wine polyphenols are also lacking, no statement can yet be made about any clinically relevant effect of these components, in either red or white wine, in terms of cardiovascular diseases. PMID- 12534099 TI - [Chromosomal mosaicism in the placenta]. AB - Chromosomal mosaicism is when two (or more) cell lines with different chromosomal complements are found within one individual. Mosaicism can be found in all tissues but may also be confined to a specific tissue. During pregnancy the aberrant cell line, often with a trisomy, may be confined to the placenta. In prenatal diagnosis, more specifically in chorionic villus sampling, this can interfere with the results and may necessitate follow-up investigation with amniotic fluid cell culture. Confined placental mosaicism can have no visible effect whatsoever on foetal development, but can also lead to serious foetal problems, depending on the chromosome involved. Uniparental disomy in the diploid cell line of the mosaicism may also have a negative effect on the foetal phenotype. In the case of the reverse situation, where the normal diploid cell line is confined to the placenta and the foetus has trisomy 13 or 18, it has been suggested that the diploid cell line might play a role in intrauterine survival; a recent study by the present authors could not confirm this view. PMID- 12534100 TI - [Physicians for ten years: a longitudinal survey of the career development of physicians who began their studies in Groningen, the Netherlands]. AB - OBJECTIVE: To analyse the career development of two cohorts of medical graduates from the University of Groningen, the Netherlands. DESIGN: Structured interviews. METHOD: For all 333 graduates who commenced undergraduate medical training in 1982 and 1983, data were collected for the period 1993-2000 with respect to resident training, income, job load, job satisfaction and scientific research. Gender differences were analysed for all of the variables. This was done by means of structured interviews as well as a strategic search in Medline and the on-line public catalogue of the University of Groningen. RESULTS: In the year 2000, 73% of the graduates had enrolled in resident training, 8% had not succeeded in finding a place. The wish to work part-time had increased strongly over the years. In general, job satisfaction was reasonable. In total 31% of the cohort had published one or more English articles or written a thesis. Gender differences were found for job load and publications. CONCLUSIONS: Ten years after graduation, medical careers had become established. Men had not succeeded in fulfilling their wishes to work part-time. Women were disadvantaged with respect to their scientific career. PMID- 12534101 TI - [Health and death of Wolfgang Amadeus Mozart]. AB - Not much is known about the medical history and cause of death of the composer Wolfgang Amadeus Mozart (1756-1791). Contrary to popular opinion, Mozart was not an exceptionally ill child and as an adult he generally enjoyed good health. At the age of 9 years he experienced a life-threatening illness, which was probably typhoid fever. On the left-hand side he had what has since become known as 'Mozart's ear', a pinna with under-development of the anthelical fold. His psychological condition gives rise to the notion that he was not exceptional, but that he responded to the events in his life in a normal manner. His terminal illness was characterised by edema of hands and feet, without any associated dyspnoea being described. The medical treatment was aggressive: enema, bloodletting on several occasions and treatment with a laxative substance. He was burried in an anonymous grave. The skull that was later ascribed to him was not his own, as no element of the denture fits with the tooth abscess he was known to have experienced. An overdose of mercury during the treatment of syphilis seems to be unlikely, as equally Henoch-Schonlein-purpura and the consequences of acute rheumatism. Trichinosis is a recent hypothesis which fits the terminal symptoms and the fact that Mozart was fond of pork. The diagnosis of his physician Closset was 'hitsiges Frieselfieber', which was possibly caused by one of the many infectious diseases prevalent at that time. PMID- 12534102 TI - ['Aai' Biemond in Propria Cures]. AB - A. Biemond (1902-1973), professor of neurology at the University of Amsterdam from 1947-1971, was the editor of the Amsterdam student weekly Propria Cures during his student years from 1921-1922. The articles he wrote for this publication were characterised by lavish baroque prose and complicated syntax with many inversions. The tone was overconfident, facetious and ironic. Themes included fellow female medical students and democracy. In later years, Biemond became an outstanding clinician and an enthusiastic teacher. He was also a participant in the doctors' resistance movement during the Second World War. PMID- 12534103 TI - [William Osler and the 'chloroforming' of men over 60: a media outcry in 1905]. AB - Sir William Osler (1849-1919) was one of the most charismatic physicians of his generation in the English-speaking world. In 1905, in a light-hearted farewell speech at the Johns Hopkins University before leaving for Oxford, U.K., he discussed the relative uselessness of men over 60. He facetiously referred to a novel in which men above this age retired for a year of contemplation, after which they were peacefully 'chloroformed'. Osler's words were reported out of context by American newspapers and he had to put up with much indignant protest. PMID- 12534104 TI - ['La medecine c'est guerir parfois, soulager souvent, consoler toujours'; in search of the origins of an aphorism]. AB - The aphorism 'La medecine c'est guerir parfois, soulager souvent, consoler toujours' is quoted rather often in the medical world. Several sources state that these words were first voiced by the famous French surgeon Ambroise Pare (1510 1590). However, further investigation suggests otherwise. It is more than likely an old saying that has possibly been roaming through medicine for centuries, but the origins of which have been lost. PMID- 12534105 TI - [Some similarities between the work of M.C. Escher and plastic surgery]. AB - At first sight there would appear to be no similarities between the work of the Dutch graphic artist M.C. Escher and plastic surgery. M.C. Escher was a gifted graphic artist who produced a large collection of work. Most of his fame is due to the works that play with symmetry, space and infinity and leave the viewer astounded. However, how Escher came to produce these works is less well known. A theory which he developed himself formed the basis of the regular plane division. It later became apparent that this theory almost completely agreed with the mathematics of plane division. Two movements (isometries) defined in mathematics, translation and rotation, are equivalent to two techniques for transferring local skin in plastic surgery, namely, advancement and transposition. Escher's performance on the plane of a sheet of paper and a plastic surgeon's performance on the plane of the skin, therefore have a similar mathematical background. Escher has visualised these mathematical rules in an unusual and artistic manner, whereas plastic surgeons apply these rules in the grace of an elastic and healing nature. PMID- 12534106 TI - [Advertising promoting self-referral in Cairo and in Los Angeles]. AB - After being a long-time resident of Cairo (Egypt) and then Los Angeles (California), the author noticed some unexpected similarities between the two cities, especially when compared to the Netherlands. In the medical field most of these similarities are related to the privatisation within the health-care system. This gives rise to a high degree of self-referral by patients and, consequently, encourages health-care providers to advertise in a sometimes aggressive manner, for example for specific hospitals, doctors, blood pressure measurement in the corner of a supermarket and even for a CT body scan. PMID- 12534107 TI - [CBO guideline 'Osteoporosis' (second revision]. PMID- 12534108 TI - [The pregnant patient with acute liver disease]. PMID- 12534109 TI - How long can employers hang on? PMID- 12534110 TI - The baby boom becomes the elder boom. PMID- 12534111 TI - Who's in charge of health? PMID- 12534112 TI - Integrating new technology without going broke. PMID- 12534113 TI - Healthier communities on the horizon. PMID- 12534114 TI - Population-based medicine: a winding road to nowhere? PMID- 12534115 TI - Still no answer for the uninsured. PMID- 12534116 TI - The health policy home fires are still burning. PMID- 12534117 TI - [Sedation in emergency medicine: a definition for emergency]. PMID- 12534118 TI - [Rapid sequence intubation in emergency: is there any place for fentanyl?]. AB - OBJECTIVES: Rapid sequence intubation (RSI) with the association of etomidate and succinylcholine is the French "Gold standard" for urgent "full stomach" endotracheal intubations. The aim of this study is to assess the fentanyl as a co induction agent to take over the sedation between the RSI and the keeping of sedation, which is a critical period in which harmful neuro-vegetatives events, and awakening signs are frequently seen. STUDY DESIGN: Randomized, double blind controlled prospective study, after acceptation by the local ethical committee. PATIENTS AND METHODS: Three groups of patients undergoing RSI in the intensive care unit and by the out-of-hospital medical team were compared: group A patients received fentanyl 3 micrograms kg-1 during RSI, before paralysis was induced. Group B patients received the same dose of fentanyl immediately after endotracheal intubation. Group C patients did not received fentanyl (control group). Outcome measures were awakening signs arrival (respiratory movements, eyes opening, spontaneous limb movements), Ramsay score assessment, and haemody namics. Attempt at intubation and vomiting incident were also measured. Discrete data were compared by chi-2 analysis, continuous data were compared with two-way analysis of variance. A p value < 0.05 was the significant threshold. RESULTS: Thirty-six patients were enrolled and completed the study. All the included patients presented awakening signs. The use of fentanyl did not prevent the recourse of other sedative medications. Ten minutes after endotracheal intubation, significant differences has been noticed for the awakening signs arrival between fentanyl groups (A: 42% and B: 36%) and control group (C: 77%). The Ramsay score evolution follows the same variation. All the patients were intubated on the first attempt, there was no vomiting incident noticed. CONCLUSION: The use of fentanyl, as a co-induction agent with etomidate and succinylcholine during RSI, allows a significant delay of the awakening signs arrival and attenuate the neurovegetative response during the minutes after endotracheal intubation after RSI, without deleterious haemodynamic effects. PMID- 12534119 TI - [Postoperative comfort in pediatric outpatient tonsillectomy]. AB - OBJECTIVE: Evaluate the comfort in children after ambulatory tonsillectomy, disregarding the surgical technique and with a systematic prescription of paracetamol and codeine. The study lasted 4 days including the day of surgery. STUDY DESIGN: [corrected] Prospective study during 5 months. PATIENTS AND METHODS: We performed a prospective study over a five-month period to evaluate the comfort in children after ambulatory tonsillectomy. Opioids were used for analgesia during surgery, morphine and propacetamol in the recovery room, and a systematic prescription of paracetamol-codeine between Day 0 to Day 3 at home. The assessment of pain was made by nurses and the family, considering 6 endpoints: spontaneous pain and when swallowing by verbal scale (0 to 4), occurrence of PONV (0 or 1), quality of sleep (0 or 1), quality of feeding (0 or 1) and play (0 or 1), combined in a global score of 0 to 12, with a score < or = 4 very satisfactory. RESULTS: 78 children were included, 49 tonsillectomy by dissection, 29 by Sluder. The score were < or = 4 at Day 0 and Day 1 in 53 children, but statistical analysis (univariate analysis) showed difference between the two surgical procedures with a global comfort score better and PONV lower with dissection procedure. CONCLUSION: The use of paracetamol-codeine after tonsillectomy offers a reliable analgesia with very satisfactory scores of comfort. Such prescription is effective in ambulatory tonsillectomy if a rigorous selection of patients is made. PMID- 12534120 TI - [Exposure of French emergency medical personnel to violence]. AB - OBJECTIVES: Evaluate the problem of violence in French EMS system and characterize assaults. STUDY DESIGN: Multicentric, descriptive, open study. PATIENTS AND METHODS: A questionnaire was given to a sample of prehospital care providers in Paris area. People were asked about assaults during their careers, typology of the assaults and consequences. Results are presented in percentage and means. RESULTS: Two hundred seventy-six questionnaires were returned. One or more assaults were recounted by 23% (61/271) of the sample (median of 8 +/- 7 years experience on the job). The injuries were bruises in 40% (17/43), wounds in 9% (4/43) and fractures in 2% (1/43). Only 4% of assaults were followed by sick leave, 15% by a complaint. After the assaults, 4% (2/45) reported having got therapy against post-traumatic stress disorder. Eighty-eight per cent reported verbal threat and 41% physical threat. Thirteen per cent (25/200) were threatened with a knife and 12% (23/200) with a gun. Only 9% (24/270) had a formal training for management of violence. CONCLUSION: Formal training in the management of violent encounters and prevention of post-traumatic stress should be developed. PMID- 12534121 TI - [Anesthesiologists in France. First results of the national survey conducted in 1999 by the French College of Anesthesiologist, The French National Society of Anesthesia and Intensive Care with the scientific support of the National Institute for Demographic Studies]. AB - OBJECTIVES: Fears related to the future of anaesthesia manpower in France have led the French College of Anaesthesiologists (Cfar) and the French Society of Anaesthesia and Intensive Care (Sfar), in scientific partnership with the National Institute for Demographic Studies (Ined), to set-up a national survey among French anaesthetists (MAR) practicing in France, to describe their demographic evolution and to analyse their professional activities. METHODS: The survey was based on a personal questionnaire, filled by each individual, approved by the National Commission on Informatics and Freedoms (CNIL). The survey was conducted in November 1998, in the 1484 hospitals, public (590) and private (894) where anaesthetics are performed, under the control of local and regional referents. RESULTS: The anaesthesiologists positions count gave a total of 9741 positions shared between 5694 in public practice (58%), 3569 in private practice (37%) and 478 in private hospitals taking part to the national health service PSPH (5%). The evaluation of the number of anaesthesiologists from the number of positions has made necessary a methodology of rectification of the survey to take in account the lack of response and the multiple sites of activity. The survey allows an evaluation of around 8876 physicians practising anaesthesia and intensive care in France at the beginning of 1999, among them 216 overseas. This census is in concordance with the count made by the Medical Council--Ordre des medecins--which published a number of 8716 anaesthesiologists in France, and 234 overseas, at the 1st January 1999, corresponding to a total of 8950. The annual demographic growth has felt from 9% per year, before 1989, to reach the level zero, in 1999. The masculinisation of the speciality is growing with a proportion of 35.7% of females, as well as ageing, the overage of age increasing from 42.8 in 1989 to 45.9 years in 1999. The pyramid of ages does not correspond to a growing population but to ageing people due to a decrease of the youngest classes. The medical density of 14.75 anaesthesiologists for 100,000 inhabitants in 1999, compared to 12.9 in 1989, is slightly above the European average, but the geographic distribution is very unequal between north and south, the large cities, centre of a university hospital, and the smaller one even if a reduction of differences is observed. The study and the analysis of professional activities bring important data to take in account side of demographic evolution. CONCLUSION: The demographic evolution must integrate non-only the reduction of the entries in the speciality, of the retirements, but also the sociological evolutions linked to the working time reduction. The solutions face to the promised shortcut of manpower consist of a reorganisation of the structures, a new definition of tasks and managements, without the possibility to avoid and adjustment of the anaesthesiologists population. PMID- 12534122 TI - [Discitis after spinal anesthesia for transurethral resection of the prostate]. AB - We described a case of discitis and meningitis following spinal anaesthesia for transurethral resection of the prostate. The patient received antibiotics for a month before surgery, because of Klebsiella prostatitis. Spinal anaesthesia was performed in L3-L4 interspace by using 22G Quincke needle. Bacteriaemia occurred during the first postoperative hours. Ten days after spinal anaesthesia, patient suffered from lumbar pain, exacerbated by vertebral percussion, and motor weakness within lower limb, which was marked on right side. MRI examination showed L3-L4 discitis with psoas abcess in regard, and epiduritis marked around L3 right spinal root. CSF examination confirmed meningitis but no bacteria was found. Antibiotics were administered over a 6 weeks period, and then patient discharged from hospital without neurological sequellae. Infectious discitis related to disk puncture during spinal anaesthesia and postoperative bacteriaemia was likely in our patient. PMID- 12534123 TI - [Unusual complication after hip surgery under spinal anesthesia]. AB - The development of an acute respiratory distress syndrome following hip surgery in elderly patients is suggestive of thromboembolism in most instances. However, we must keep in mind the possibility of rarer complications, which can remain undiagnosed because they are hidden by prominent abnormal behaviours, which can develop following any type of anaesthesia. We report the case of a patient who developed a confusion following an orthopaedic surgery under spinal anaesthesia; this confusion concealed a penetration syndrome resulting from accidental inhalation of a dental crown. Because this patient was old and had previously developed chronic lung disorders, we selected a spinal anaesthesia for performing the surgery; these underlying respiratory disorders worsened the clinical consequences of the inhalation. The dental crown was removed under general anaesthesia with spontaneous ventilation using a bronchoscope after an unsuccessful attempt with a fibrescope due to the size of the foreign body. PMID- 12534124 TI - [Multitruncular block at the elbow for a major hand trauma for prehospital care]. AB - Upper limb peripheral nerve blocks offer many advantages but they are not widely used in prehospital care. We report the case of a multitroncular block at the elbow made by the emergency department team out of the hospital after a major hand trauma. A successful axillary brachial plexus block was done two hours later, without any problem. This case report allows us to discuss about different analgesia techniques useful in prehospital care. PMID- 12534125 TI - [Fulminant coma: think hyperammonemia and urea cycle disorders]. AB - The authors report the case of 14-year-old boy admitted for acute coma without neurological focal symptom. The only relevant finding was the death of one uncle after a coma in the year 1992. This coma was associated with an ammonia blood level of 344 mumol l-1 and it rapidly lead to cerebral death despite a symptomatic treatment. The diagnosis of hereditary ornithine transcarbamylase deficiency was confirmed by liver biopsy in the immediate post-mortem period. The authors recommend the measurement of blood ammonia in every coma without diagnosis, whatever patient's age. PMID- 12534127 TI - [Difficulty of withdrawing epidural catheter due to a knot]. PMID- 12534126 TI - [Hemostasis disorder in a patient with hemangioblastoma of the cerebellum and polycythemia]. PMID- 12534128 TI - [Clostridium difficile small intestinal involvement occurring after total colectomy]. PMID- 12534129 TI - Imported viral haemorrhagic fever with a potential for person-to-person transmission: review and recommendations for initial management of a suspected case in Belgium. AB - Viral haemorrhagic fevers are caused by a wide range of viruses. There are 4 types of viruses well known to spread from person to person and able to cause nosocomial outbreaks with a high case fatality rate: an arenavirus (Lassa fever and more exceptionally the Junin and Machupo virus), a bunyavirus (Crimean-Congo haemorrhagic fever) and the Filoviridae (Ebola and Marburg viruses). So far there have been only a limited number of imported cases of viral haemorrhagic fever in industrialized countries. In recent years an increasing number of outbreaks of filovirus infections have occurred in Africa and in 2000 5 cases of Lassa fever were brought from Sierra Leone to Europe. Therefore European physicians should consider the possibility of a viral haemorrhagic fever in an acutely ill patient just returning from Africa or South-America with fever for which there is no obvious cause. Such patients should be questioned for risk factors for viral haemorrhagic fever. Using universal precautions for handling blood and body fluids and barrier nursing techniques there is little risk that if a patient with viral haemorrhagic fever arrives in Belgium there will be secondary cases. PMID- 12534130 TI - Questionnaires are better than laboratory tests to screen for current alcohol abuse or dependence in a male inpatient population. AB - OBJECTIVE: To assess the diagnostic performance of the CAGE and AUDIT (Alcohol Use Disorder Identification Test) and its derivatives, and laboratory tests for screening alcohol abuse or dependence in a male medical hospital population. DESIGN: A diagnostic cross-sectional prevalence study. SETTING: Three general hospitals and one university hospital. PATIENTS: All male patients older than 18 years admitted to the hospitals, during a period of 6 weeks, were consecutively included in the study (N = 233). MEASUREMENTS: Calculation of diagnostic measurements with 95% CI and ROC curves for different scores of CAGE, AUDIT and derivatives, laboratory tests and % Carbohydrate Deficient Transferrin (CDT), using DSM-III-R as the reference standard, derived from the CIDI. RESULTS: A current diagnosis of alcohol abuse or dependence was found in 29 medical male inpatients, representing 12.4% (95% CI: 8.6-17.5). Ten of these (4.2%) fulfilled criteria of alcohol abuse and 19 (8.2%) the criteria of alcohol dependence. Laboratory tests are useless as screening tools with sensitivities between 10% (%CDT) and 52% (GammaGT). Only the Fiveshot questionnaire seems to yield reasonable diagnostic parameters at the recommended cutpoint of > or = 2.5 with a sensitivity of 79.3% and a specificity of 87.7%. CONCLUSIONS: With a prevalence of 12.4%, our results are similar with other published studies for alcohol abuse and dependence according DSM criteria. The AUDIT as well as the Fiveshot seems to have the best diagnostic properties in this male medical inpatient population, and regarding to brief interventions, these questionnaires can be used as screening instruments as well as opportunities to talk about alcohol problems with admitted patients. PMID- 12534131 TI - Rising digoxin serum levels and intoxication despite digoxin withdrawal in an elderly patient: a case report. PMID- 12534132 TI - Pasteurella multocida in peritoneal dialysis: a rare cause of peritonitis associated with exposure to domestic cats. AB - Pasteurella multocida is a rare cause of peritonitis in peritoneal dialysis patients with only 10 cases reported in the literature so far. All cases were observed in patients with close contact with cats, usually with a direct puncture of the dialysis tubing. We report a case of Pasteurella multocida peritonitis in a patient maintained under continuous cycling peritoneal dialysis (CCPD), who had frequent and close contact with cats. Patients should be made aware of this potential complication and advised to keep domestic animals away from the location of their peritoneal exchanges. PMID- 12534133 TI - Management of the patient with chronic renal failure in the evidence based era. AB - Chronic renal failure is a growing problem with an increasing number of patients suffering from loss of kidney function. The morbidity and mortality of these patients is much higher than that of the general population. The patient with chronic kidney disease, even with only a moderate level of renal failure, falls in the highest risk category for cardiovascular disease. A proper evaluation and management of these patients is necessary to prevent further loss of kidney function, to prevent cardiovascular diseases and to manage the co-morbid conditions associated with renal failure and the complications due to renal failure. The therapies available for these purposes in adult patients with chronic renal disease not (yet) in dialysis, are presented in this article, with emphasis on blood pressure management, antiproteinuric therapies, correction of renal anaemia, approach of lipid disorders. A timely referral to a nephrologist is also shown to be important. PMID- 12534135 TI - Treatment of erectile dysfunction. AB - The evaluation and treatment of Erectile Dysfunction has evolved dramatically over the last 10 years thanks to the improvement of our understanding of the physiology of erection and the development of effective drugs to be taken "on demand" before intercourse. In addition, Erectile Dysfunction is now clearly recognized as a medical disorder. It is beyond the scope of this article to go into details about the physiology and pathophysiology of the sexual cycle and the different diagnostic procedures for evaluating erectile disorders. Rather, we will concentrate on the therapeutic options for managing erectile disorders after a brief review of the most recent concepts of erection physiology and diagnosis. PMID- 12534134 TI - Genetic polymorphisms and clinical practice: the example of osteoporosis. AB - Over the past decades epidemiological research of so-called "complex" diseases, such as diabetes, osteoporosis, cancer, and cardiovascular disease, has identified a number of risk-factors for these common age-related disorders. Examples include obesity for diabetes, low bone mineral density for osteoporosis, and smoking for cancer and cardiovascular disease. Some of these risk factors are used in clinical practice, for example to identify subjects-at-risk and for monitoring treatment of patients with these diseases. More recently, a novel class of risk factors, genetic polymorphisms, has gained considerable interest in this respect. This is mostly due to the Human Genome Project which has identified every human gene and also uncovered a plethora of polymorphic variants of these genes which embody the genetic risk factors. Genetic risk factors are expected to find applications in early identification of subjects-at-risk, prediction of response-to-treatment, and development of novel therapeutic options. We will here discuss some developments in the field of the genetics of osteoporosis. PMID- 12534136 TI - Management of primary osteoporosis. AB - Although there is a great need for better therapeutic approaches to the patient who presents with a fracture, osteoporotic fractures will remain a condition that is more amenable to prevention than treatment. Hormone replacement therapy (HRT) is still considered by many the mainstay for the prevention and the treatment of posrmenopausal osteoporosis. However, there are several controversies regarding HRT, especially the duration of treatment and the risks/benefits ratio. Recent studies have challenged the assumption that HRT conveys real long-term beneficial effects. Raloxifene or other "selective estrogen receptor modulators" (SERMs) should progressively replace HRT in elderly women. Bisphosphonates have demonstrated a clearcut efficacy in the treatment of osteoporosis. Alendronate and risedronate have been the most extensively studied bisphosphonates under randomized controlled trials conditions. Both agents can reduce the risk of vertebral and hip fractures by one-fourth to one-half. However, oral bisphosphonates are not without gastro-intestinal toxicity and strict adherence to constraining therapeutic schemes is mandatory. Intermittent treatments are already in use. Weekly alendronate is as efficient as daily therapy and improves treatment compliance. Newer more potent bisphosphonates, such as oral ibandronate or intravenous zoledronic acid, will allow much less frequent administration. The anti-fracture efficacy of yearly zoledronic acid infusions is thus currently tested. On the other hand, bone-forming agents, such as daily subcutaneous injections of teriparatide (rhPTH 1-34) offer exciting perspectives for the treatment of severe osteoporosis despite the complexity of such therapy. PMID- 12534137 TI - The endless challenges of end-of-life care. PMID- 12534138 TI - The defendant physician's deposition: fighting back--at last! PMID- 12534139 TI - Physician warn patients on CT screening. PMID- 12534140 TI - HIPAA after October 15 ... what now? PMID- 12534141 TI - Professional liability insurance rates--some history & solutions. AB - The medical malpractice business, as with the property and casualty industry as a whole, is cyclical in nature. We have experienced peaks and valleys since the mid 1970s. History would tell us that profitability can be restored in time. Many in the professional liability insurance industry think that a return to stability is possible by 2004. The two most critical factors in restoring this profitability may be adequate pricing and tort reform--time will tell the story. PMID- 12534142 TI - Pain relief at the end-of-life: a clinical guide. AB - Pain at the end-of-life is usually treatable, but most dying patients are under treated and die in unnecessary pain. This brief overview will serve to describe the problem of pain at the end-of-life, define the relevant ethical, medical, scientific, and societal issues, and present an optimal pain management plan for this vulnerable and important population. The most important factor is for physicians to make pain control a matter of paramount importance in the care of dying patients. PMID- 12534143 TI - Relieving pain: what are today's ethical and legal risks? AB - Many physicians have poor understanding about the symptoms of terminal illness. There is also surprisingly little data about treating symptoms compared to treating disease. Subsequently pain and other forms of suffering are often under treated, despite the availability of effective interventions. Fear of retribution is often coupled with the moral notion that aggressive pain treatment may cause premature death. Legal and ethical obligations may compete but an appreciation of both is needed to inform effective palliative care. PMID- 12534144 TI - Questions & answers about hospice: a guide for Missouri's physicians. AB - Hospice provides multidisciplinary care to dying patients with and without cancer. Most adults would prefer to be cared for in their home or that of a family member. This guide provides answers to the questions most commonly asked of physicians. Its goal is to facilitate a better understanding of what hospice does, who is eligible, physician roles, and how physicians can use hospice to help their patients. PMID- 12534145 TI - Spirituality and end-of-life care. AB - As dying patients adjust to the irreversible nature of their illness, their needs and focus of care changes. Spiritual issues may become a central concern for them, and addressing these issues can be key to relieving suffering. Physicians, unfortunately, have little training in this area and are often uncomfortable discussing spirituality. In this article, we address the role of spirituality in end-of-life care, and discuss a format for spiritual assessment. We hope this will encourage more comprehensive patient-centered, end-of-life care. PMID- 12534146 TI - Finding health in folklore, herbs and supplements: the good, the bad & the ugly. Part II-- The ugly. AB - The dark 'bad' side of herbal and supplement use is not only in their direct adverse effects or toxicity, but in their potential for interactions with a variety of agents commonly used by mainstream practitioners. In the next issue, Part III--The Ugly--will focus on the distorted image of these hazardous herbal drifters. PMID- 12534147 TI - Effects of nicotinic acid and lovastatin in combination with cholestyramine in renal transplant patients. AB - We assessed the short and long-term efficacy and safety of the lipid lowering drugs, nicotinic acid, and the HMG-CoA reductase inhibitor, lovastatin, in combination with cholestyramine in four renal transplant patients. Combination therapy with cholestyramine failed to lower the TC and LDL to the levels recommended by the NCEP guidelines. PMID- 12534148 TI - Rationale for the epidermal growth factor receptor as a target for cancer therapy. AB - OBJECTIVES: To provide information regarding tumor-specific molecules and signaling pathways responsible for regulation of tumor growth, and review the development of novel targeted therapies that can block the epidermal growth factor receptor (EGFR) at biologically active and less toxic doses compared with standard chemotherapy. DATA SOURCES: Research articles. CONCLUSIONS: The identification and understanding of novel molecular targets has led to the development of therapeutic agents specific to these targets. Clinical investigations involving EGFR inhibitory agents are being completed. Studies have shown single-agent efficacy and efficacy in combination with chemotherapy and/or radiation therapy. IMPLICATIONS FOR NURSING PRACTICE: Nurses are involved in the administration of targeted therapies. They serve to ensure effective patient education, compliance, and early intervention for toxicities. Knowledge regarding the impact of signaling pathways and the goals of targeted therapy will maximize treatment outcomes for cancer patients. PMID- 12534149 TI - The epidermal growth factor receptor as a novel target for cancer therapy: case studies and clinical implications. AB - OBJECTIVES: To update the success and obstacles using therapies targeted to the epidermal growth factor receptor including IMC-C225, ZD1839, and OSI-774. DATA SOURCES: Research articles, textbooks, clinical protocols, case studies. CONCLUSIONS: Ongoing studies show promise using monoclonal antibodies and small molecule tyrosine kinase inhibitors in the treatment of non-small cell lung cancer and other malignancies. Such therapies can be delivered with limited toxicity to the host, and initial studies have shown single-agent efficacy and efficacy in combination with chemotherapy and/or radiation therapy. IMPLICATIONS FOR NURSING PRACTICE: Nurses are directly involved in the delivery of these novel therapies and the management of associated toxicities. They serve to coordinate treatment protocols and help to ensure patient understanding and compliance. PMID- 12534150 TI - Overview of clinical trials with epidermal growth factor receptor inhibitors in advanced non-small cell lung cancer. AB - OBJECTIVES: To provide an overview of the clinical development of ZD1839 and other epidermal growth factor receptor (EGFR) inhibitors in the treatment of advanced non-small cell lung cancer (NSCLC). DATA SOURCES: Research articles and abstracts. CONCLUSIONS: Increased understanding of cancer biology has led to the development of multiple targeted agents to evaluate clinically in the treatment of various cancers. The EGFR inhibitors appear promising in the treatment of NSCLC. ZD1839 has shown activity and symptom and quality-of-life improvements in two large phase II trials. Ongoing and future trials will continue to explore the optimal use of ZD1839 and other EGFR inhibitors in the treatment of NSCLC. IMPLICATIONS FOR NURSING PRACTICE: Several investigational EGFR inhibitors are likely to become commercially available over the next few years. Nurses should be knowledgeable regarding these agents to provide appropriate patient and family education and to assure safe drug delivery. PMID- 12534151 TI - Even more opinions on PVC and broader lessons. AB - A Scientific Committee of the European Commission has addressed possible toxicity concerns of plasticised poly(vinylchloride) in certain medical devices and has reached conclusions that are different from those adopted by some other organisations. This article discusses the basis for these opinions. PMID- 12534152 TI - Current trends in molecular diagnostics. AB - Rapid progress in molecular diagnostics is expected to fundamentally change health care. This review of some of the latest innovations covers developments in polymerase chain reaction (PCR), several nonPCR methods, and biochips and nanotechnology, which have accelerated advances in the analysis of nucleic acids and proteins. The greatest impact is likely to be in the integration of diagnosis and therapy and the development of individualized treatment. PMID- 12534153 TI - Hydrogel adhesives for wound-care applications. AB - This review of pressure-sensitive adhesives used in wound dressings describes the properties of recently developed hydrogel adhesives. The findings of a study comparing the adhesion and moisture vapour transition rates of the hydrogel adhesives with some existing wound-dressing products are reported. PMID- 12534154 TI - The technological edge. AB - Surface-engineered bearings for hip prostheses have been developed that produce fewer wear particles and extend the life of the prosthesis. PMID- 12534155 TI - Design, development and optimisation using computational fluid dynamics. AB - Interest in more advanced drug delivery systems has increased with an acceleration in the discovery and development of novel therapeutic macromolecules for targeted applications. Computational fluid dynamics is a design tool that allows designers of these and other products to evaluate different designs rapidly and cost-effectively. PMID- 12534156 TI - Determining resistance to environmental stress cracking in luer fittings. AB - Environmental stress cracking (ESC) is a phenomenon associated with disposable plastic products. This article describes new methods to determine ESC in luer fittings. The findings of two experimental studies are reported. PMID- 12534157 TI - Adverse event management during clinical investigations. AB - The requirements are changing for documentation and evaluation of adverse events that occur during clinical investigations of medical devices. This article discusses important amendments to adverse event definitions contained in a draft international standard and methods for improving the process of managing these types of events. PMID- 12534158 TI - Performance evaluation and the IVD Directive. AB - Performance evaluation is a new procedure for many in vitro diagnostics manufacturers. This article describes how to conduct a trial. PMID- 12534159 TI - Standards in the 21st century. Standardising new medical technologies, Part I. PMID- 12534160 TI - The importance of low-fluorescence adhesives. AB - Adhesives that exhibit low auto-fluorescence are important in medical diagnostic devices where fluorescence detection is used to determine an analyte. This article reports on the advantages of these adhesives in the manufacture of products such as microfluidic devices and kinase assays and discusses customization opportunities. PMID- 12534161 TI - Early warning technology. PMID- 12534162 TI - Rotary converting technology. PMID- 12534163 TI - Tagging medical devices. PMID- 12534164 TI - Labour-management forums and workplace performance. Evidence from union officials in health care organizations. AB - Many health care workplaces are adopting more cooperative labour-management relations, spurred in part by sweeping changes in the economic environment that have occurred over the last decade. Labour-management cooperation is seen as essential if health care organizations are to achieve their valued performance objectives. Joint labour-management committees (LMCs) have been adopted in many health care workplaces as a means of achieving better industrial relations. Using data from a sample of Canadian union leaders in the health care sector, this paper examines the impact of labour-management forums and labour climate on employee and organizational outcomes. Research results suggest that labour climate is less important in predicting workplace performance (and change in workplace performance) than is the number of LMCs in operation. However, labour climate is found to be at least as important in predicting union member satisfaction (and change in member satisfaction) as is the wide adoption of LMCs in operation. These findings are consistent with the notion that the greater use of LMCs is associated with augmented workplace performance (and a positive change in workplace performance), notwithstanding the contribution of the labour climate in the workplace. PMID- 12534165 TI - Variation in coding influence across the USA. Risk and reward in reimbursement optimization. AB - Recent anti-fraud enforcement policies across the US health-care system have led to widespread speculation about the effectiveness of increased penalties for overcharging practices adopted by health-care service organizations. Severe penalties, including imprisonment, suggest that fraudulent billing, and related misclassification of services provided to patients, would be greatly reduced or eliminated as a result of increased government investigation and reprisal. This study sought to measure the extent to which health information managers reported being influenced by superiors to manipulate coding and classification of patient data. Findings from a nationwide survey of managers suggest that such practices are still pervasive, despite recent counter-fraud legislation and highly visible prosecution of fraudulent behaviors. Examining variation in influences exerted from both within and external to specific service delivery settings, results suggest that pressure to alter classification codes occurred both within and external to the provider setting. We also examine how optimization influences vary across demographic, practice setting, and market characteristics, and find significant variation in influence across practice settings and market types. Implications for reimbursement programs and evidence-based health care are discussed. PMID- 12534166 TI - From home, to market, to headquarters, to home. Relocating health services planning and purchasing in New Zealand. AB - Health sector restructuring has been in vogue, but no country has engaged in as much health sector restructuring as New Zealand where, in a decade, there have been four different public health sector structures. This article discusses New Zealand's four structures with an emphasis on relocating the critical functions of health care planning and purchasing, and on the development of the present district health board system. The four structures include: an area health board system (1989-1991) with planning and purchasing located at "home" in local areas and closely aligned with service provision; a competitive internal market system (1993-1996) which separated planning and purchasing from service provision; a centralised system with a "headquarters" controlling planning and purchasing (1997-1999) while maintaining the distance from provision; and the district health board system currently under development (1999-) which sees purchasing and planning sent home again to regions and linked closely with service provision. The present system entails the devolution of considerable responsibility to the local level, within a framework of strong central government control. Based on New Zealand's experience, the article notes that all but the market structure appear to have provided an adequate environment for effective health care planning and purchasing. PMID- 12534167 TI - Non-profit multi-hospital organizations: challenges and prospects. AB - This study seeks to determine the relative importance of factors non-profit hospital administrators rely on in their decisions to join a non-profit multihospital organization (MO) and their assessments of an MO in satisfying these motives. A related objective of the study is to determine whether or not the administrators of different types of hospitals (i.e. general vs specialty, member of a national vs non-national MO and church-affiliated vs non-church affiliated) differ in their judgements. The analytical framework of the importance-performance technique is used in analyzing the data gathered from the top administrators of a nation-wide sample of hospitals in the USA. Results and implications of the study are discussed. PMID- 12534168 TI - Evaluating self-managed integrated community teams. AB - After briefly describing self-managed integrated community teams, the authors explore potential and actual methods of evaluating their structures, processes and outcomes. Primary health care staff in three comparable sites were studied using non-participant observation, interviews, focus groups and questionnaires. After describing the fieldwork, the authors examine integrated team structures, which are characterised by a large number of barriers that integrated teams face. Processes surrounding different working practices are explored next. Ways of unifying health care professional practice in integrated teams are suggested using evidence from both the literature and fieldwork. Outcomes that emerged after one year of the new teams' lives are discussed in detail. The difficulty in establishing acceptable outcomes, especially the validity and reliability of outcome measures, is considered. Throughout, the positive and negative aspects of integrated teams emerging from the fieldwork are compared and contrasted with issues in the literature. Finally, recommendations are made to help strengthen integrated teams in the UK. PMID- 12534169 TI - [Humanitarian action: its requirements, its stakes]. AB - Humanitarian action is developing in an increasingly changing and troubled world context. Organizations are subject to more and more complex requirements due to ethical, legal, technical, financial, media-related, and political considerations. They must deal with higher and higher stakes such as the emergency-development trade-off, the need for professionalization, and the revival of governmental humanitarian action. The articles in this special issue deal with all these topics and more. In most cases the authors are players in the field of humanitarian action from non-governmental, intergovernmental, international and national organizations. There are also academics, politicians, and civilians. PMID- 12534170 TI - [Bioforce Development Institute approach: professional training center for humanitarian workers]. AB - The Bioforce Development Institute has been providing training, orientation, and expertise since its founding by Doctor Charles Merieux in 1983. Non-governmental organizations (NGO) require that applicants for humanitarian missions exhibit not only motivation and adaptability but also specific skills. Training is necessary to prepare personnel prior to departure. In addition NGO must hire salaried personnel and develop human resources with the same efficiency as any private sector company. Personnel working for International Relief Organizations (IRO) must be competent in project management. To meet these growing needs, Bioforce offers training to logistics and administrative managers working in international humanitarian organizations. PMID- 12534171 TI - [European Center for Humanitarian Health (CESH). Providing operational know-how directly applicable in crisis situations]. AB - Founded in 1997 at the initiative of Charles Merieux, the European Center for Humanitarian Health Care (French acronym, CESH) is a Public Interest Organization comprising 7 members, i.e., Lyon 1 Claude Bernard University, Aix-Marseille II University of the Mediterranean, the Public Hospital System of Lyon, the Public Hospital System of Marseille, the French Army Health Corps, the Merieux Foundation, and the National School of Veterinary Medicine in Lyon. The CESH is a multifunctional resource dedicated to providing education, information, and research in the field of humanitarian action. The objectives of the CESH's Educational Program are to increase awareness of the complexity of humanitarian action and teach the principles and methods necessary to integrate experienced teams already in the field. Courses including a three-week study program sanctioned by an Interuniversity Degree in Public Health and several 2-to-3-day training modules are open to all humanitarian actors including field workers and decision-makers, health-care professionals, and governmental or other administrative agents. Thanks to a diverse faculty with academic, humanitarian, training, civilian and military backgrounds and to a wide-ranging curriculum, the CESH helps to promote communication and to open up closed pathways of cooperation between the different groups involved in humanitarian action. The module entitled "Humanitarian Assistance and Civilian and Military Cooperation" exemplifies the effort of the CESH to generate collaborative behavior among civilian and military actors. The CESH's Research Program is aimed at providing practical tools for decision-making in the field. The web site of the CESH [http://cesh.univ lyon1.fr] presents the mission and activities of the Center as well as information on course enrolment and humanitarian action. PMID- 12534172 TI - [Management of a cholera epidemic by a humanitarian organization]. AB - Far from disappearing with modern progress, cholera outbreaks have never been more common than in the last decade. Because of its close relationship with poverty, malnutrition, poor access to health care, warfare, and sometimes natural disaster, cholera is a major priority for action by doctors from humanitarian organization. In this article a brief overview about cholera is followed by an account of one non-governmental organization's approach to management of a cholera outbreak. The author describe the circumstances surrounding the alert and the fact-finding mission focused on documenting the presence of an epidemic, measuring its magnitude, and predicting its outcome. The next phase of management involved response planning with development of a control strategy and raising money for intervention, which is often difficult for epidemics not reported in the media. The control modalities were then deployed in the field including assisting patient management, preventing transmission, training personnel, supervising activities, coordinating tasks performed by different actors, and finally program evaluation. This experience shows that successful cholera management programs can be conducted in a professional manner by teams composed essentially of volunteers under the supervision of a knowledgeable staff able to adapt general control techniques to the specific conditions encountered in the field. PMID- 12534174 TI - [Water supply for civilian populations in crisis and post-war situations: example of Mitrovica city (Kosovo) in 1999-2000]. AB - After reviewing the effects of water shortage on sanitation, the author describes minimum water requirement in terms of both quantity and quality during emergency situations and recovery periods. This article also gives general guidelines for water treatment to cover vital needs and provide adequate sanitation to the victimized community. To illustrate his point, the author cites the water management program implemented by the humanitarian action branch of an industrial group in Mitrovica, Kosovo in 1999 and 2000. PMID- 12534173 TI - [Humanitarian intervention, example of the French Civil Defense as a government organization]. AB - The French Civil Defense is a governmental organization dedicated to saving life and alleviating suffering after disasters in France and foreign countries. It provides aid during the crisis phase of the emergency based on detailed assessment of needs. The French Civil Defense organizes a range of emergency services including medical care, water supply, and search and rescue in cooperation with local and foreign teams. PMID- 12534175 TI - [Practical management of nutritional and food problems in crisis situations]. AB - This article deals with one of the most consistent problems arising during crisis situations, i.e., food shortage. The author first presents the international context conducive to natural or man-made famine. He emphasizes the importance of early detection and the need for an evaluation phase using public health survey skills to determine the objectives of relief. He then describes strategies for management of alimentary and nutritional requirements in famine-stricken populations in function of the different types of malnutrition encountered. The article ends with a presentation of preventive measures that must be implemented in association with crisis management. PMID- 12534176 TI - [Military Bioforce, example of governmental humanitarian action]. AB - Governmental humanitarian action is an old concept in France, that is a topic of current interest. To conduct humanitarian action, the French government has deployed various facilities. The military biological risk team was established to control epidemics, provide expertise during epidemiological emergencies, and conduct mass vaccination programs. The author describe the missions carried out by the biological risk team during the 1990s as an illustration of governmental humanitarian action. PMID- 12534177 TI - [Epidemiological surveillance during humanitarian emergencies]. AB - Humanitarian emergencies are complex situations that usually occur after warfare or social upheavals. They result in a dramatic increase in morbidity and mortality in the affected population. Organizations that undertake management of humanitarian emergencies must be prepared to provide comprehensive services covering all aspects of public health including preliminary evaluation, mass vaccination, water and sanitation, food supply and nutrition, shelter, medical care, control of communicable diseases, personnel training, and logistic coordination. Epidemiological surveillance is an integral part of a full-service approach to humanitarian action that should be included in the preliminary evaluation. Epidemiology is a powerful tool for detecting any health problems that might further jeopardize the victim population. By allowing measurement of the impact of humanitarian action, epidemiological data also provides a means of verifying that the victims are benefiting from aid operations. Implementation of a surveillance system should be given high priority during the start-up of a humanitarian operation. The system must have the simplicity and flexibility necessary to adapt changing, unstable situations as well as high sensitivity necessary for early detection of epidemic outbreaks. PMID- 12534178 TI - [Access for all to quality drugs]. AB - The implementation of essential drugs policies and widespread use of generic products in humanitarian programs has ensured access to medication for poverty stricken populations. However rigorous drug selection according to origin is necessary to guarantee quality. A drug is more than a mixture of chemical components, it depends on a complex balance that can be affected by numerous factors. To be beneficial to the patient, a drug must be manufactured, purchased, distributed and administered in a strictly professional and responsible manner. An obvious danger for drugs such as antibiotics is that use of poor quality products producing insufficient bioavailability will promote development of microbial resistance. This could become a worldwide public health problem with particularly dramatic consequences for the treatment of tuberculosis and retroviruses. PMID- 12534179 TI - [Emergency preparedness based on coherent integration of crisis response resources: workshop on mass casualty management in Bethlehem in June 1999]. AB - In June 1999, the Humanitarian Action Division of the Foreign Affairs Department organized a training course on the mass casualty management within the framework of religious festivities related to the Bethlehem 2000 project. This initiative was undertaken at the request of the Palestinian Authority and the General Consulate of France in Jerusalem. Palestinian and French specialists in the field worked together in three workshops devoted to rescue, search and first aid; medical outposts and triage; and emergency and surgical care in referring hospitals. PMID- 12534180 TI - [Workshop of military reconstructive surgery in Conakry May 1999: example of civilian-military cooperation in responding to medical-surgical crisis emergencies]. AB - During the period from 1998 to 1999, civil wars broke out in number of west African countries including Liberia, Sierra Leone, and Guinea Bissau. Due to the situation in surrounding countries, Guinea Conakry was forced to accept nearly 650000 refugees whose presence represented a major risk for the socio-political stability of the country. International organizations and NGOs condemned the atrocities inflicted on civilian populations by the children serving as soldiers in the RUF rebels organizations of Sierra Leone and Liberia. These attacks included murders, gang rapes, abduction of children and young people, and mutilation of extremities of people of all ages ranging from infants to elderly. Treatment of mutilation victims requires the availability of facilities for surgical treatment and prosthetic fitting in Guinea Conakry. The humanitarian action division of the French Foreign Affairs Department and the NGO Handicap International decided to provide specialized training in the management of mutilation injuries to surgical groups in hospitals of Guinea and Sierra Leone. The program consisted in a workshop on reconstructive surgery for war-related injuries to allow optimal prosthetic fitting for reinsertion of mutilation victims into society. PMID- 12534181 TI - [Relations between military health services and humanitarian organizations--the point of view of the International Red Cross]. AB - The classic definition of humanitarian action is any operation conducted for the purpose of helping to reduce human suffering. This concept can be expanded to include a notion of prevention. This expanded definition has opened a wide range of applications for humanitarian assistance. Military personnel are frequently involved in relief operations after natural disasters. Military logistic support is a key factor for coping with the massive needs of victims. However military intervention in humanitarian operations during wartime poses complex problems. The ICRC intervenes during military conflict and has, for several years, expressed its concern over the growing involvement of military forces in humanitarian operations. These considerations provide the backdrop for analysis of the relationship between Military Health Corps Services and the ICRC. The purpose of this article is to evaluate the need to make a clear-cut distinction between political and humanitarian objectives and thus between the actors associated with these goals. In this regard, the role of Military Health Corps Services will be presented in function of the principles underlying any humanitarian program. This article will also discuss areas of agreement and cooperation between the ICRC and Military Health Corps Services. PMID- 12534182 TI - [The law, an indispensable instrument for humanitarian action]. AB - In this article, the author describes, in layman's terms, the legal framework for international humanitarian operations. He explains a number of complex and intricate principles used in these situations. He acknowledges the burden that legal considerations place on humanitarian organizations but also demonstrates that legal expertise is an indispensable tool in the provision of humanitarian services. PMID- 12534183 TI - [Questions concerning humanitarian action]. AB - Although development of humanitarian action is rooted historical events, the dynamics behind today's international relief organizations can only be understood within the context of the modern world. Relief organizations are currently confronted with major challenges and paradoxes. The challenges include the need to enhance professionalization and standardization of assistance operations and exposure to greater risks. The paradoxes involve the need to implement complex, highly publicized programs in a simplistic manner and problems involved in managing the complex relationship between relief workers and victims, tainted with the almighty powers of the actors. PMID- 12534185 TI - [Daily risks in the devaluation of the image of humanitarian action]. AB - After restating the definition, purpose, and role of humanitarian assistance, the author emphasizes the dangers that misuse for political considerations represents for the image of humanitarian action. While recognizing that technical expertise is important for effective provision of aid, the author stresses that humanitarianism requires more than technology. He also warns that collecting the funds necessary for relief operations must not lead to commercial-style marketing of humanitarian assistance. Similarly he cautions that the targets for humanitarian action should not be determined by television pictures. In conclusion the author advocates responsible administration of humanitarian programs. PMID- 12534184 TI - [Humanitarian action threatened by standardization]. AB - The author analyses the new international context in which humanitarian action is being undertaken. He raises the problem caused by the diverging objectives of impartial, neutral humanitarianism and politically motivated actions that implement strategies of prevention and conflict resolution. He reviews the criticism that humanitarian has come under in recent years and that has resulted in establishment of codes of conduct. However he points out the threat that the concepts of control and "jurisdiction" over humanitarian action represent and analyzes discrepancies between minimal standards and universal principles. The article concludes with a presentation of an alternative solution based on the "Quality" platform being developed by several French NGOs. PMID- 12534186 TI - [Impact of war on civilian population health. Republic of the Congo, December 1998-February 2000]. AB - Congo Brazzaville was racked by civil war between 1998 and 2000. Fighting forced a large part of the population to flee from the capital, Brazzaville. Starting in 1999 many people began coming back to the city in dire physical and psychological condition. With little or no press coverage, this return went largely unnoticed by the general public. Volunteers from Medecins Sans Frontieres were present not only to provide assistance and care to these war victims but also as to bear witness so that this tragedy should not be forgotten. By means of epidemiological surveys in the field, workers were able to paint a "picture of suffering" and to document the violence and deprivation inflicted mainly on the civilian population. This article summarizes the findings of this investigation. PMID- 12534187 TI - [A war against civilians. Reflections on humanitarian practice in Congo Brazzaville (1998-2000)]. AB - This article beckons readers to reflect on the extreme acts of violence inflicted on populations when civil warfare degenerates into war against civilians. The authors raise the issue of political, media, and humanitarian choices determining the response of industrial countries to crisis situations in economically challenged countries. They advocate an approach based on the "assist and testify" principle of Medecins Sans Frontieres. PMID- 12534188 TI - [Continual evolution of government humanitarian action]. AB - The author gives a brief historical overview of the administrative structures that have progressively been set up for management of governmental humanitarian action. He stresses that governmental humanitarian programs are no longer limited to a strictly administrative functions but now include a new altruistic interface involving consultation with non-governmental organizations. However he also underlines that governmental humanitarian action is still carried out not only as an act of human assistance but also as an act of foreign policy. He ends with a presentation of the recently created Humanitarian Action Commission (HAC) at the French Foreign Office. PMID- 12534189 TI - [Political dimensions of humanitarian action: an analysis framework]. AB - The author proposes an approach borrowed from political sociology and centered around Conflict/Mediation dialectic pair as a basis for objective analysis of the relationship between Humanitarian Action and Politics. PMID- 12534190 TI - [Civilian-military coordination]. AB - Current humanitarian emergencies create complex, mutidimensional situations that stimulate simultaneous responses from a wide variety of sources including governments, non-governmental organizations (NGO), United Nations agencies, and private individuals. As a result, it has become essential to establish a coherent framework in which each actor can contribute promptly and effectively to the overall effort. This is the role of the United Nations Office for the Coordination of Humanitarian Affairs. Regardless of the circumstances and level of coordination, cooperation and collaboration between humanitarian and military personnel, it is necessary to bear in mind their objectives. The purpose of humanitarian action is to reduce human suffering. The purpose of military intervention is to stop warfare. The author of this article will discuss the three major obstacles to civilian-military coordination (strategic, tactical, and operational). Operations cannot be conducted smoothly and differences cannot be ironed out without mutual respect between the two parties, an explicit definition of their respective duties and responsibilities, a clear understanding of their cultural differences, and the presence of an organization and facilities for coordination and arbitrage by a neutral referee. PMID- 12534191 TI - [Civilian-military actions]. AB - This article presents the concept of civilian and military cooperation (CIMIC) including its definition, origin, goals, and implementation. The author details the value and role of CIMIC in responding to crisis situations. To illustrate his point, the author includes a short description of CIMIC during the Kosovo crisis. PMID- 12534192 TI - [Which relationships between civilian-military actions(CMA), civilian and military cooperation (CIMIC)and non-governmental organizations(NGO)?]. AB - This article questions the existence of "governmental humanitarianism" in opposition to "non-governmental" humanitarianism. The authors report the growing implication of civilian and military actors representing governments in responding to crises and express their concern that this tendency could ultimately lead to use of humanitarian action for diplomatic and economic purposes. They stress that unlike non-governmental organizations, state-run humanitarian agencies are subject to political, economic, and geographic factors. In the context of increasingly frequent application of the CMA doctrine, the authors analyze the role of non-governmental organizations in the field during post-crisis reconstruction. They underline the growing problem of organizing bilateral and multilateral assistance operations. They restate the hope expressed by many humanitarian organizations for a return of the rule of law and conclude on the need to develop complementary activities in the field. PMID- 12534193 TI - [Conclusion: challenges and stakes in humanitarian action in the XXIth century]. AB - The authors in the special issue devoted to humanitarian action use their own experience, analysis, and thought as a basis for identifying the challenges and stakes facing humanitarian action in the twenty-first century. They present their thoughts on the relevance of humanitarian intervention, on the need to combine emergency aid with development assistance, and on the development of governmental humanitarian action. In today's world there is a compelling obligation for communities with resources and means to undertake humanitarian action in the name of human dignity. Adopting a code of conduct, respecting minimal standards, and evaluating the outcome are now universally accepted principles, but the modalities of applying these principles are still subject to discussion especially with regard to standards and accreditation. The importance of adequate training and preparation for humanitarian action has also been established. Currently most humanitarian operations involve numerous actors from different organizations that may be national or international as well as governmental, intergovernmental, or non-governmental. A major portion of this issue is devoted to the relationship between military forces and humanitarian organizations. The need to coordinate action in the field is emphasized. Humanitarian actions are increasingly complex operations carried out in fast-changing situations by numerous players. As a result it has become more and more important that actors in the field know each other in order to work together efficiently and thus better reach their common goal of relieving and preventing human suffering in accordance with ethical principles. PMID- 12534194 TI - [CO2 laser surgery for the treatment of vocal cord carcinomas in early stages (T1 T2)]. AB - BACKGROUND: The standard treatment for patients with early vocal cord carcinoma has been radiotherapy. In recent years, encouraging results with CO2 laser surgery has changed our treatment strategy in selective cases. During the past four years at the Rabin Medical Center we have treated patients with carcinoma of the vocal cord using CO2 laser surgery. PURPOSE: We conducted a retrospective study to investigate the results of the Head and Neck Surgery department with CO2 laser treatment for squamous cell carcinoma of the vocal cord during early stages (T1, T2) without radiotherapy. MATERIALS AND METHODS: Twenty one patients with carcinoma of the vocal cords in early stages were treated with CO2 laser. Nineteen patients suffered from stage T1 carcinoma located in one vocal cord, three of these patients suffered from involvement of the anterior commissure and one patient also had involvement with the anterior part of the second vocal cord. Two patients with carcinoma in stage T2, one with involvement of the ventricular band and one with limited movement of the vocal cord. The follow-up period for the patients ranged from one month to four years. RESULTS: Only one patient in stage T1 developed local recurrence after half a year, no patients died during the follow-up. CONCLUSION: Careful patient selection with endoscopic staging also during the biopsy, removal of the carcinoma in free border, strict follow-up and stroboscopic examination are essential to secure good results in the treatment of CO2 laser for early stages of vocal cord carcinoma. PMID- 12534195 TI - [Cerebral paraplegia due to blunt head injury]. AB - Cerebral paraplegia due to traumatic brain injury is a very rare condition. We present a case study of a man who sustained a blunt head injury, sub-dural hematoma of the falx and dorsal paraplegia. Eventually, he recovered almost completely neurologically and functionally. PMID- 12534196 TI - [Acute rectal bleeding from malignant stromal tumor]. AB - Stromal tumors of the rectum are rare. The most common symptoms are rectal bleeding, pain, constipation and obstruction. A case report of a 61 years old female admitted with acute bleeding due to malignant stromal tumor of the rectum is described. Upon admission the clinical and laboratory findings necessitance blood transfusion. Preoperative assessments revealed a large tumor located in the rectal wall with extension to the adjacent tissues. The histologic features of needle biopsy was compatible with gastrointestinal stromal tumor. Abdominoperineal excision of the rectum was performed. The definitive histological examination showed a gastrointestinal stromal tumor (GISTs), spindle cell type with mucosal invasion, vast necrosis and highly mitotic activity. The tumor cells coexpressed CD34 and smooth muscle actin and were negative for staining desmin and S-100 protein. Adjuvant chemotherapy was given. The patient died 7 months after the operation due to widespread metastases. We decided to describe a rare malignant stromal tumor of the rectum with uncommon clinical presentation. PMID- 12534197 TI - [Repair of anorectal fistulas using fibrin glue tissue adhesive--preliminary experience in 15 patients]. AB - PURPOSE: The goal of this study was to evaluate whether commercially produced fibrin sealant can be used for the treatment of complex anorectal fistulae. METHODS: Fibrin glue was used for patients with primary and recurrent ano-rectal fistulae, but patients with Crohn's disease or AIDS were excluded. The procedure was performed under anesthesia in the operating room. The primary and secondary openings were identified using blue dye. The secondary opening and the tract of the fistula were curetted, and 4 ml of fibrin sealant were injected through the secondary opening until fibrin sealant was seen coming from the primary opening. The patients were discharged on the next day following the procedure, and were followed-up closely in the outpatient clinic. RESULTS: Fifteen consecutive patients underwent one trial of fibrin glue injections. The average follow-up period was 4 months. Overall, eleven of the 15 patients (73.3%) had successful closure of their fistulae. The healing rates for intersphincteric, transsphincteric and suprasphincteric were 50%, 77.7% and 100% respectively. Fibrin glue treatment was effective in five out of the 6 patients (83.3%) who were operated on for recurrent fistulae. Of the 4 patients in whom the treatment failed, the mean time for recurrent symptoms was 4 weeks. No patient suffered from fecal or gas incontinence and all of the patients returned promptly to routine life. CONCLUSION: Sealing the tracts with fibrin glue is a unique mode for the treatment of ano-rectal fistulae, and our preliminary results with this technique are promising. The procedure is safe and easy to perform, and can be offered as an alternative to conventional surgery. Most importantly, this method minimizes the risk of fecal incontinence and the discomfort of prolonged wound healing. PMID- 12534198 TI - [SCIM--spinal cord independence measure (version II): sensitivity to functional changes]. AB - Until recently, the functional ability of patients with spinal cord lesions (SCL) was assessed by standardized scales designed for various disabilities. However, these scales have either a relatively low sensitivity to changes in the functions that are most important for SCL patients or a limited suitability for a specific SCL subgroup. To counter this problem the team of the Spinal Department of Loewenstein Rehabilitation Hospital developed the Spinal Cord Independence Measure (SCIM), which is specific for SCL patients, adjusts for disadvantages of earlier scales, and is user-friendly. It was found to be reliable and more sensitive to functional changes in SCL patients than the Functional Independence Measure (FIM), the most often used disability scale today. A second version (SCIM II) was constructed with improved phrasing of some of the components. It too, was found reliable, even more than the original version for certain functions. The present study examined the sensitivity of the SCIM II to changes in function in SCL patients compared to the FIM. Twenty-six patients with SCL underwent sequential SCIM II and FIM examinations during hospitalization for rehabilitation. A high correlation was found between the total scores of the two scales (r = 0.915; p < 0.0001). The mean change in function score from the first to the last examination was significantly larger with the SCIM II than with the FIM (p < 0.04), and the rate of detection of functional change was usually higher with the SCIM II. The advantage of the SCIM II over the FIM in detecting functional changes was evident in areas in which the two scales differ substantially. These results support the validity of the SCIM II. Studies with larger groups in different countries and cultures are still needed before the scale can be applied on an international basis. PMID- 12534200 TI - [Life and death at the tip of the tongue--the power of words]. AB - The names of various common diseases have an etymological source derived from the names and revelations of scholars, scientists, physicians and famous figures from the near and distant past. This article unveils some of the mystery, the tales and the etymology behind those diseases. PMID- 12534199 TI - [High incidence of resistant pathogens in community acquired bacteriuria from patients in the Jerusalem area with lower urinary tract infections]. AB - Lower urinaty tract infections are among the most common bacterial infections encountered in community practice. The diagnosis is made on a clinical basis, and the treatment provided is therefore empirical. In order to characterize the uropathogens and their antimicrobial susceptability, we have screened posive urine cultures which were isolated from patients in Jerusalem area with symptoms suggesting lower urinary tract infection. We found that, similar to other communities, E. coli is still the most common uropathogen (66.1%) in the Jerusalem area. The antibiotic susceptibility was (for women and men respectively) 29.8%, 25.1% for ampicillin, 51.6%, 41.1% for cephalexin, 86%, 78% for nitrofurantoin, 67%, 54% for TMP-SMX, 62.9%, 49.3% for amoxicillin-clavulanic acid, 92.8%, 77.2% for gentamicin, 88.8%, 64.4% for cefuroxime and 91.4%, 71.7% for ciprofloxacin. According to the above antimicrobial susceptability nitrofurantoin can be recommended for empirical treatment of lower urinary infections in women while in men, non E Coli pathogens were more common and more effective drugs such as cefuroxime and quinolones were needed. PMID- 12534201 TI - [Tissue microarray technology--a new and powerful tool for the molecular profiling of tumors]. AB - Tissue Microarrays (TMA) are the products of a new technology that offers rapid and simultaneous analysis of up to 1000 different archival samples. Sections from TMA blocks can be used for all different types of in situ tissue analysis including immunohistochemistry and in situ hybridization. The technology helps to implement the vast knowledge that was discovered by the human genome project and facilitates the molecular understanding of various benign and malignant processes. In this article we describe the process of TMA construction and review the main studies that applied the technology to characterizing tumors and precancerous lesions. PMID- 12534202 TI - [Adherence to antipsychotic medication in schizophrenia patients]. AB - Antipsychotic agents are the mainstay of treatment for schizophrenia. However, almost 50% of schizophrenia patients exhibit non-compliance with antipsychotic drug therapy. This in turn may account for high relapse rate, reduced quality of life and increased burden on society. Identification of factors that may mediate adherence to treatment is therefore a primary clinical challenge. This paper reviews the literature regarding medication adherence among schizophrenia patients, with an emphasis on four groups of mediating factors: illness-related, patient-related, treatment-related and environmental-related factors. In addition, we describe a modified health-belief model that may facilitate better understanding of empirical findings. Therapeutic interventions to encourage better treatment compliance are discussed. PMID- 12534203 TI - [Non-freezing cold injury in soldiers]. AB - Non-freezing cold injury (NFCI) is an injury of the hands or feet resulting from exposure to wet conditions and temperatures just above freezing, typically found in soldiers. NFCI is due to microvascular endothelial damage, stasis and vascular occlusion. At first, the tissue is cold and anesthetic, progressing to hyperemia in 24-48 hours. Hyperemia is accompanied by an intense painful burning sensation as well as blisters, redness, and possibly, ulcerations. NFCI management raises frustration in both medical officers and commanders. Most authorities are not aware of or remain unimpressed with the severity of NFCI, nor do they realize that it produces lifelong symptomatology. The following review of the available literature attempts to clear up some issues regarding the definition, pathogenesis, symptoms and preventative measures available in NFCI, including our own experience. PMID- 12534205 TI - [New approaches in the treatment of short stature]. AB - Over the past years, the efforts to improve the final height of children with short stature has yielded new modalities of therapy. New types of injection devices, mainly ready-to-use injection pens with multi-dose vials of reconstituted growth hormone (GH), have made the treatment technically easier and less stressful for the patients. A new type of slow-release GH injected once or twice a month has recently been studied. In addition, several types of synthetic GH-secretagogues were examined, with the advantage of being introduced by the oral or nasal route. The recognition of the pivotal role of estrogens in skeletal maturation and subsequent growth arrest, was the basis for the use of aromatase inhibitor in a pioneering study, with promising results. We have concluded the review with an update on the benefit of GH therapy in children with normal short stature, based on 12 studies that provided data on the final height of these children. PMID- 12534204 TI - [Coronary artery ectasia: a therapeutic dilemma]. AB - Coronary artery ectasia (CAE) is an abnormal dilatation of the coronary artery wall. The growing number of coronary angiographies and other invasive cardiologic procedures increased the documented incidence of CAE. There is no consensus about the etiology, prognostic significance and morbidity related to this phenomenon. Atherosclerosis is most probably the main cause of primary or idiopathic ectasia. In addition, it is postulated that chronic exposure to vasodilatatory substances, and therapeutic angioplastic procedures may mediate secondary ectasia. CAE is associated with increased coronary morbidity such as coronary spasm, dissection and thrombus formation. However, its relative contribution to coronary morbidity remains unclear. This uncertainty underlies the current dispute regarding the appropriate management and treatment of patients with CAE. Further studies including prospective therapeutic trials are needed to provide answers to these pending complex questions. PMID- 12534206 TI - [Retinopathy of prematurity--risk factors]. AB - Retinopathy of prematurity (ROP) was first described by Terry in 1942. ROP is considered a multifactorial disease. Low gestational age, low birth weight and oxygen therapy are recognized as risk factors for this condition. Other risk factors including multigestational pregnancy, white race, sepsis, NEC, BPD, intraventricular hemorrhage, lung maturation, steroid treatment, blood transfusions and light exposure were identified by multiple studies. We aim to review these studies in order to identify the independent risk factors for the development of ROP. The reviewed studies confirm that low birth weight, low gestational age, prolonged oxygen treatment and blood transfusions are statistically significant risk factors for the development of ROP. The incidence of all stages of ROP is similar for Caucasian and black infants, although the occurrence of threshold ROP was found higher in the Caucasian group. No relationship was demonstrated between light exposure and the development of ROP. The studies reviewed show decreased frequency and severity of ROP in neonates of mothers who had received antenatal steroid therapy. The findings concerning the influence of postnatal steroid treatment on the incidence of ROP are controversial. PMID- 12534207 TI - [Health Geographic Information System (HGIS)--a tool for health planning and epidemiology]. AB - The development of the Health Geographic Information System (HGIS) concept, associated to the wide availability of adapted software applications, has offered an invaluable tool to Public Health Professionals. However its use is still too often limited to the graphic presentation of data or to structural planning, neglecting its main advantages. The HGIS has proven to be a potent tool for risk assessment, decision making, intervention evaluation and health planning. In spite of all the obstacles, the transformation of geographic information systems (GIS) into an epidemiological tool adapted to the health system (HGIS) can now be done in a relatively short span of time, with a minimal investment in terms of budget and manpower. The use of an outsourcing policy enables to limit the operational running costs in health organizations such as the Health Ministry, Hospitals or Sick Funds. PMID- 12534208 TI - [New approaches to immune against hepatitis C virus]. AB - Hepatitis C virus represents a global health problem as some 170 million individuals are infected worldwide. Studies on virus replication and pathogenesis are hampered by the lack of a reliable cell culture system. In addition, the high rate of mutations reflected in viral antigen variability complicates attempts to prepare an efficient vaccine against HCV. Recently some significant strides were made towards resolving some of the above-mentioned difficulties. The importance of several structural antigens i.e. HVR1 and CD81 was demonstrated with regard to the infection mechanism and as structures that may serve to immunize against the virus. A promising new approach of using recombinant DNA vaccines instead of protein vaccines is discussed. PMID- 12534209 TI - [ISMH--International Society of Medical Hydrology and Climatology]. PMID- 12534210 TI - [The Israeli Medical Association--the first years]. PMID- 12534211 TI - Male age, host effects and the weak expression or non-expression of cytoplasmic incompatibility in Drosophila strains infected by maternally transmitted Wolbachia. AB - In Drosophila melanogaster, the maternally inherited endocellular microbe Wolbachia causes cytoplasmic incompatibility (CI) in crosses between infected males and uninfected females. CI results in a reduction in the number of eggs that hatch. The level of CI expression in this species has been reported as varying from partial (a few eggs fail to hatch) to nonexistent (all eggs hatch). We show that male age in this host species has a large impact on the level of CI exhibited and explains much of this variability. Strong CI is apparent when young males are used in crosses. CI declines rapidly with male age, particularly when males are repeatedly mated. Wolbachia from a Canton S line that was previously reported as not causing CI does in fact induce CI when young males are used in crosses, albeit at a weaker level than in other D. melanogaster strains. The strain differences in CI expression are due to host background effects rather than differences in Wolbachia strains. These results highlight the importance of undertaking crosses with a range of male ages and nuclear backgrounds before ascribing particular host phenotypes to Wolbachia strains. PMID- 12534212 TI - Nucleotide polymorphism at the Atmyb2 locus of the wild plant Arabidopsis thaliana. AB - DNA variation was studied in a 2.2 kb region of the regulatory gene Atmyb2 using 20 ecotypes of Arabidopsis thaliana and one accession each of Arabis gemmifera and Arabidopsis himalaica. Nucleotide diversity (pi) in the region was 0.0027, which was lower than for other loci in A. thaliana. The MYB domain of the Atmyb2 gene (pi = 0.0036) had a larger variation than the non-MYB region (pi = 0.0013). Tajima's test and Fu and Li's test did not give a significant result. In contrast to the low level of polymorphism, the degree of divergence of the Atmyb2 region was higher between A. thaliana and A. gemmifera (K = 0.0730) than for other loci. The MYB domain (K = 0.0436) had smaller divergence than the non-MYB region (K = 0.0939). The HKA test detected significant discordance in the ratio of polymorphism to divergence in some comparisons. The pattern of low polymorphism and high divergence, which is mainly observed in the non-MYB region of the gene, is inconsistent with the neutral mutation theory. Strong purifying selection after establishment of A. thaliana and a species-specific adaptive process could be invoked to account for this pattern of polymorphism and divergence of Atmyb2. PMID- 12534213 TI - The mutational rate of Drosophila viability decline: tinkering with old data. AB - In the first 25 generations of his classical mutation accumulation experiment, T. Mukai estimated a large rate of early linear decay for the relative viability of Drosophila melanogaster chromosome II (delta MII = 0.004). Mukai forced through zero the regression of viability decline on generation number, but it has recently been shown (Fry, 2001) that a similar decline (delta MII = 0.006) is obtained from unforced regression even if generation 32 instead of generation 25 (whose validity has been questioned) is included. We show that, from the perspective of the whole long-term experiment. it is hard to decide up to which generation viability can be considered to decline linearly. Depending on this decision, and on whether or not the regression is forced through the origin, very different estimates are obtained. Furthermore, the particular behaviour of the lines used as control suggests that they could have been different from the remaining lines at the beginning of the experiment, and casts doubts on the adequacy of a forced regression. Estimates from the linear unforced regression (delta MII = 0.011) or from the linear term in a quadratic unforced regression (delta MII = 0.001) are very different. The data fit both models very well, and the choice between them should be based on biological grounds. PMID- 12534214 TI - Inbreeding depression and heterosis in a subdivided population: influence of the mating system. AB - We investigate the joint effects of gene flow and selfing on the level of inbreeding depression, heterosis and genetic load in a subdivided population at equilibrium. Low gene flow reduces inbreeding depression and substantially increases heterosis. However, in highly self-fertilizing populations, inbreeding depression is independent of the amount of gene flow. When migration occurs via pollen, consanguinity of the reproductive system could have a negative influence on subpopulation persistence, in contrast to the case of isolated populations. However, with only seed migration, genetic load and heterosis depend mildly on the mating system. From an evolutionary point of view, we reach two main conclusions: first, outcrossing is selected for if gene flow is low; second, intermediate levels of gene flow could promote mixed mating systems, especially when migration occurs through pollen. PMID- 12534215 TI - Studies of mitochondrial DNA, allozyme and morphometric variation in a house mouse hybrid zone. AB - An unusual chromosomal hybrid zone of the house mouse, Mus musculus domesticus, exists in Upper Valtellina, Northern Italy, consisting of four Robertsonian (Rb) races and the standard (all-acrocentric, or 2n = 40) race, all hybridizing freely within 10 km2. The hybrid zone in Valtellina provides an excellent opportunity to study the role of Rb fusions in reproductive isolation and speciation. This hybrid zone has already been well studied for the distribution of Rb fusions and the fertility of hybrids, but in order to understand the dynamics of the zone, a basic understanding of the origin and genetic similarity of the chromosomal races is necessary. This paper presents the results of three different methods of measuring genetic differentiation: multivariate analysis of morphological traits and analyses of allozyme variation and mitochondrial DNA sequences. The standard race is clearly distinguishable from the three Rb races by all three methods, but the Rb races are not distinguishable from one another. This provides strong evidence for our previous suggestions that the well-established Rb races in Valtellina are closely related, and that the standard race was introduced into the valley more recently from a distant source. The fact that the Rb races are indistinguishable is also consistent with our hypothesis that a within-village speciation event involving two of the races (Hauffe & Searle, 1992) was a recent occurrence. The low level of allozyme heterozygosity among the Rb races suggests that these populations are the products of at least one bottleneck. The present article substantially extends earlier studies and provides the first detailed morphological and molecular analysis of this complex hybrid zone. PMID- 12534216 TI - Molecular linkage map of Einkorn wheat: mapping of storage-protein and soft-glume genes and bread-making quality QTLs. AB - Two molecular maps of Triticum monococcum L were produced and integrated. The integrated map includes a total of 477 markers, 32 RFLPs, 438 AFLPs, one morphological (soft glume (Sog)) and six storage-protein markers, and covers 856 cM. The trait Sog with the recessive allele sog maps to linkage group 2S. Probably, this is the T. monococcum homologue of Tg and Tg2 in hexaploid and tetraploid wheats, respectively. Loci coding for seed storage proteins were allocated to chromosomes 1L (HMW GLU1,2 and Glu1), 1S (LMW GLU6,7, LMW GLU1-4, omega GLI1-4, gamma GLI5 and Gli-1) and 6L (alpha/beta GLI7-14). Parameters related to bread-making quality (SDS sedimentation volume, specific sedimentation volume (SSV) and total protein content) were studied in one of the two populations. A QTL that is consistently present across environments was detected for SDS sedimentation volume and for SSV. The position of the QTL on chromosome 1S was in close agreement with the map positions of storage-protein loci. A second QTL was mapped on chromosome 5. For protein content, two significant QTLs were mapped to linkage groups 1 and 5. PMID- 12534217 TI - QTL mapping for growth and carcass traits in an Iberian by Landrace pig intercross: additive, dominant and epistatic effects. AB - Results from a QTL experiment on growth and carcass traits in an experimental F2 cross between Iberian and Landrace pigs are reported. Phenotypic data for growth, length of carcass and muscle mass, fat deposition and carcass composition traits from 321 individuals corresponding to 58 families were recorded. Animals were genotyped for 92 markers covering the 18 porcine autosomes (SSC). The results from the genomic scan show genomewide significant QTL in SSC2 (longissimus muscle area and backfat thickness), SSC4 (length of carcass, backfat thickness, loin, shoulder and belly bacon weights) and SSC6 (longissimus muscle area, backfat thickness, loin, shoulder and belly bacon weights). Suggestive QTL were also found on SSC1, SSC5, SSC7, SSC8, SSC9, SSC13, SCC14, SSC16 and SSC17. A bidimensional genomic scan every 10 cM was performed to detect interaction between QTL. The joint action of two suggestive QTL in SSC2 and SSC17 led to a genome-wide significant effect in live weight. The results of the bidimensional genomic scan showed that the genetic architecture was mainly additive or the experimental set-up did not have enough power to detect epistatic interactions. PMID- 12534218 TI - [Prion diseases: current views]. AB - This is a review of the problem. The above diseases represent a peculiar type of fatal neurodegenerative diseases of man and animals provoker of which is prion--a low molecular anucleinic protein resistant to inactivation. Human prior diseases include fatal familial sleeplessness, Kreutzfeldt-Jakob disease, kuru, Gerstmann Streussler-Sheinker's syndrome, etc. Prion proteins possess a characteristic capacity to change a tertiary conformation resulting in the formation of a pathological form of protein having infectious properties. Pathogenesis, morphological and clinical manifestations as well as principles of diagnosis and treatment of these diseases are discussed. PMID- 12534219 TI - [Prognostic significance of p53, HER-2/neu, Ki-67 and VEGF expression in chondrosarcomas]. AB - Expression of the mutant protein-suppressor of tumor growth p53 and the receptor of epidermal growth factor of type II--HER-2/neu may serve as prognostic factors for patients with chondrosarcoma of malignancy grade II. Combined expression of these proteins in the tumor tissue correlates with shorter 5-year survival of recurrence-free patients. The data obtained may be used for prognosis of chondrosarcoma course and choice of adequate therapy. PMID- 12534220 TI - [The role of epithelial antigens in diagnosis and staging of breast cancer]. AB - The study of different epithelial antigen expression has been performed in 183 breast cancers. In 73 patients regional lymph nodes were studied as well. Panepithelial antigen Egp-34 (Mab HEA-125) was expressed in 100% of primary tumors and metastases. Antigen MUC-1 (Mab ICO-25) was identified in 93% of breast cancers. Monomorphic type of expression in tumor cells was typical for Egp-34, MUC-1 being in certain cases expressed as a proportion of cells. Additional immunohistochemical study of regional lymph nodes with Mab to Egp34 and MUC-1 provides a 10% increase in the rate of breast cancer metastases detection compared to histological examination alone. The carcinoembryonic antigen (CEA) was useful in identification of metastases only in CEA-positive breast cancers. PMID- 12534221 TI - [Immunohistochemical characteristics and a degree of differentiation of urinary bladder cancer]. AB - 17 cases of severe dysplasia and urinary bladder intraepithelial carcinoma, 15 cases of invasive well differentiated urothelial carcinoma, 14 cases of moderately differentiated and 10 cases of poorly differentiated carcinoma were studied immunohistochemically. The role of metalloproteinases in urinary bladder carcinoma is established. Correlation between proliferative activity, protease expression and the degree of tumor differentiation is found. Viral DNA of human papilloma of 16/18 types in severe dysplasia and carcinoma in situ was revealed by hybridization in situ method. PMID- 12534222 TI - [Histological and molecular-biological markers of malignancy in different variants of thyroid papillary carcinoma]. AB - Material for this study from 61 patients (thyroid papillary carcinoma) is shown to represent a heterogenic group of tumors differing from each other morphologically, molecular-biologically and histogenetically. The most malignant were long-cell and B-cell variants and the least malignant--a follicular variant. 81% of the tumors had neuro-endocrine differentiation and their malignant potential was higher compared to tumors without this differentiation. PMID- 12534223 TI - [Immunohistochemical examinations of dyskeratosis and exocervical neoplasia in gynecological pathology]. AB - An immunohistochemical study was carried out with monoclonal antibodies to estrogen receptors (Ers), Ki-67, BCL-2, P53 and human papillomaviruses (HPV) of normal cervix (7 cases), cervical leukoplakia (5 cases), CIN I (7 cases) and CIN II-III (6 cases). Correlations were also investigated between hyperestrogenemia and molecular changes in the exocervix. About two thirds of patients with leukoplakia and precancerous changes had clinical signs of hyperestrogenemia which led to proliferation of estrogen-sensitive parabasal cells. Due to neoplastic epithelial transformation the number of cells with Ki-67, P53 and HPV positive reactions in the nuclei increased, while Ers and BCL-2 expression decreased. Because active proliferative parabasal cells are needed for HPV replication we consider hyperestrogenemia as a factor stimulating development of neoplastic changes in the uterine cervix. PMID- 12534224 TI - [Morphometric characteristics (assessment of ploidy) of the degree of differentiation of uterine body adenocarcinoma]. AB - 53 histological slides obtained from 4 patients aged 35-68 years with diagnosis of uterine carcinoma without invasion (16 patients) and tumor invasion into uterine muscular membrane (24 patients) were retrospectively analyzed. On the basis of morphometry and ploidometry of 2989 nuclei on the image analyzer Imager CG (Russia) with a computer program Avtan-San, a complex of diagnostic criteria characterizing the grade of malignancy of uterine tumors was obtained. Differential diagnostic ploidometric characteristics of 4 degrees of tumor progression are described. With the process intensification the amount of genetic material in tumor cell nuclei increase 1.7-fold, proliferative activity 2.2-fold, number of polyploid cells--1.5-fold. These data specify the degree of differentiation of uterine body adenocarcinoma and help to plan treatment policy for patients with uterine carcinoma. PMID- 12534225 TI - [Characteristics of tumor cell nucleolar apparatus in stomach carcinoma]. AB - The nucleolus in gastric carcinoma cells is changing in the course of tumor progression. The number of argentophylic granules in the nucleoli depends on tumor cell proliferative and protein-synthetic activity: rise with adenocarcinoma differentiation change from high to low degree. PMID- 12534226 TI - [Morphological characteristics of gastrointestinal lesions in patients after total colectomy for colon polyposis]. AB - 11 patients with juvenile polyposis and 4 patients with familial adenomatous polyposis after total colectomy entered the study. Long-term follow-up with endoscopic examination and multiple biopsies in these patients showed high probability of polyps in preserved regions of the gastrointestinal tract. Therefore, regular prophylactic endoscopy, morphological examinations and biopsies of endoscopically normal mucous membrane are justified. Such policy is recommended especially for patients with familial adenomatous polyposis. PMID- 12534227 TI - [A comparative morphometric analysis of cytograms of invasive ductal and invasive lobular breast cancer]. AB - The most informative for differential diagnosis parameters (surface and nucleus polarization) were determined on the basis of morphometric analysis of 42 cytograms of invasive duct cancer (IDC) and 39 cytograms of invasive lobular breast cancer (ILC) using the computer analyzer MEKOC-C. Combination of these two parameters allowed the authors to elaborate a decisive rule for differential diagnosis of these two cancers (the mistake is reduced to 8.6%). The analysis of 8 informative morphometric nuclear parameters has shown that in ILC the nuclei are more monomorphic, less in square and perimeter, nucleoli are less numerous than in IDC. The study gives objective criteria for preoperative differential cytological diagnosis between IDC and ILC. PMID- 12534228 TI - [Primary culture of prostatic adenocarcinoma: a model for studies of stromal epithelial interactions]. AB - Primary culture of prostatic adenocarcinoma is obtained from surgical material after radical prostatectomy. Typing of the obtained culture with antibodies to vimentin and cytokeratins has shown that the proportion of basic cell components- stromal and epithelial--in vitro correlates with such in the initial tumor tissue. In cultivation in different media active migration and proliferation of all cell types in the presence of embryonal serum and suppression of the stromal component in the medium where the serum was replaced for bovine hypophysis extract were observed. A comparative immunohistochemical analysis registered a release into culture of alpha SMA-positive myofibroblasts and active expression of TGF beta-1 in the medium containing serum. Design of dynamic mixed cell systems may serve a convenient model for investigation of stromal-epithelial interactions and their changes in cancer progression. PMID- 12534229 TI - [A retrospective morphological analysis of endomyocardial biopsies of myocarditis patients]. AB - A repeated retrospective analysis of histoultrastructure of endomyocardial biopsies of patients with diagnosis of myocarditis and with various life span after manifestation of the first disease symptoms. The most unfavorable sign of prognosis is advanced myocardiocyte necrosis, alterative changes and the degree of interstitial cell reaction. Death of cardiocytes due to apoptosis was not typical for myocarditis and was observed only in several patients from the group with long survival. A new type of cardiomyocyte death--extrusion of the nucleus from the cell--was found in the group with low survival. PMID- 12534230 TI - [Morphological features of atherosclerotic plaques of internal carotid artery sinus (biopsy data from carotid endarterectomy)]. AB - A histological study was made of 140 atherosclerotic plaques in the sinus of the internal carotid artery (ICA) removed at carotid endarterectomy. New formed thin walled vessels of various diameter having the structure of arteries, blocking arteries and, presumably, arteriolo-venous anastomoses were found in 112 plaques. The majority of the vessels were oriented along the ICA this indicating blood circulation parallel to that in ICA. It is assumed that deficiency of blocking arteries results in the development of intraplaque hemorrhages from thin-walled vessels. PMID- 12534231 TI - [Secondary autoimmune syndrome and immunomodulating lymphotropic therapy in acute pancreatitis]. AB - Immune status and condition of regional lymphoid tissue were studied in 28 patients with various forms of acute pancreatitis untreated with immunomodulating therapy and in 12 patients receiving combined therapy including lymphotropic regional therapy with T-activin. Acute pancreatitis patients were found to have aquired immunodeficiency, secondary autoimmune syndrome and reaction of toxic immune complexes the severity of which decreased under the action of T-activin therapy. PMID- 12534232 TI - [Intestinal metaplasia of mucosal epithelium of the urinary bladder]. AB - 4 cases of urinary bladder intestinal metaplasia are reported. One adenoma and one adenocarcinoma of intestinal type were discovered. Immunohistochemically, expression of meconial intestinal antigen beta 1 MA in the cytoplasm and on the apical surface of goblet-like cells was observed. PMID- 12534233 TI - [Epidemiology, pathogenesis and pathological anatomy of leptospirosis]. AB - The review gives information on epidemiology, pathogenesis and pathologic anatomy of leptospirosis as well as the role of endotoxin in selective damage to the liver and kidneys, causes of death. PMID- 12534234 TI - [Reorganization of DNA replicating system induced by 5-fluorodeoxyuridine and caffeine as the basis for radiosensitivity in human calls]. AB - Treatment of diploid human fibroblasts with 5-fluorodeoryuridine (FUdR) and caffeine results in the increase in cellular radiosensitivity in terms of survival and chromosomal aberrations, on the one hand, and in radioresistant DNA synthesis (RDS), on the other hand, i.e. rather mimics those in mutant cells from patients with AT, XPII, Down syndrome, and others. A study was made of the autoradiographic length of simultaneously active adjacent replicon clusters. After incubation of diploid human cells with FUdR (10(-6) M, 6 h), this parameter was shown to reduce by two-fold, remaining unchanged upon 5 Gy irradiation. In contrast, after incubation of the cells with caffeine (2 mM, 30 min), this parameter was longer, compared to that in intact cells; upon 5 Gy irradiation the values remained almost the same as in the control. A possible relation of the data to the cellular radiosensitivity and RDS in the cells incubated with FUdR and caffeine is discussed. PMID- 12534235 TI - [Effects of cationic vectors complexed with plasmid DNA on the phagosome-lysosome fusion in murine peritoneal macrophages and J744 macrophages]. AB - Polyethylenimine (PEI) and cationic polypeptides complexed with plasmid DNA are the most efficient nonviral vectors for gene therapy. It is believed that endocytosis is the major pathway for cell entering by PEI/DNA or cationic peptides/DNA complexes. Effects of plasmid DNA complexed with PEI, poly-L-lysine (PLL), poly-D-lysine (PDL) and polyarginine (PA) on the phagosome-lysosome fusion (P-LF) were studied in murine peritoneal macrophages and J774 macrophages. Cationic polypeptide PLL can be hydrolysed by cellular peptidases, but its stereoisomer, PDL, cannot be split by these enzymes. PEI, PDL, and PA have been shown to inhibit P-LF. PLL showed a low effect on the P-LF. On the basis of these studies, we assume that lysosomotropic agents able to change functions of lysosomes in the cell may affect transfection efficiency and thus be used for gene therapy. PMID- 12534236 TI - [A new human cellular protein AUP1. I. In vitro interaction of AUP1 with adenoviral proteins E4ORF3 and E1A]. AB - The 11-kDa product of adenovirus early region 4 (E4) open reading frame (ORF) 3 participates in many processes occurring in infected cell, including post transcriptional steps in late viral gene expression and viral DNA synthesis. In addition, E4ORF3 from adenovirus type 5 (Ad5) displays the features of a viral oncoprotein. It initiates focal transformation of primary rat cells in cooperation with Ad5 El genes and confers multiple additional transformed properties on E1-expressing cells. Biochemical details of E4ORF3 activities in these processes are not well understood. A large body of evidence indicates that its lytic and transforming functions are mediated by physical interactions with viral and cellular components involved in DNA transcription and repair, as well as by host cell factors that regulate the integrity of nuclear multiprotein complexes known as PML oncogenic domains (PODs). In this study we have employed the two-hybrid screen in yeast to isolate human cDNAs encoding for E4ORF3 interacting proteins. Among 15 positive clones five cDNAs encode for a cellular protein called AUP1. In vitro-binding assays demonstrated that AUP1 fused to glutathione S-transferase (GST) specifically binds to E4ORF3 from Ad5, Ad9 and Ad40 generated in a coupled transcription-translation system, whereas no interactions was observed with ORF3 from Ad12. Interestingly, GST-AUP1 interacted also specifically with in vitro translated Ad5 E1A proteins. Regions involved in the Ad5 E4ORF3/AUP1 interaction in vitro map to the central part of E4 protein and the carboxy-terminal region of AUP1, while E1A binds to an amino-terminal segment of the cell protein. Taken together, these studies indicate that AUP1 may represent a cellular target of both adenovirus E4ORF3 and E1A proteins. Additional studies are currently under way to confirm the significance of these interactions in living cells in vivo. PMID- 12534237 TI - [A new human cellular protein AUP1. II. cDNA cloning, genomic organization of Aup1 gene ans preliminary characterization of human AUP1 protein]. AB - We report here the cloning of human cDNA of Aup1 gene (ancient uniquitous protein 1), its genomic organization and preliminary characterization of human AUP1 protein. Genomic length of the human Aup1 gene is composed of 12 exons and 11 introns spanning 3047 bp. A single open reading frame of 1230 bp starts with the first ATG from nucleotide 5630 of the genomic sequence, in accordance with the Kazak rule, and stops with TGA on nucleotide 8500 of the genomic sequence AC005041.2. The 3 noncoding region has a 152 bp length followed by poly(A)tail. All exon-intron junctions conform to 5' donor and 3' acceptor consensus. Exon and intron sizes range from 69 to 187 bp, and from 180 to 262 bp, resp. We have cloned a full-length human Aup1 cDNA. For preliminary characterization of human AUP1 protein, we cloned the full-length human Aup1 cDNA in pcDNA3 vector. In vitro transcription/translation of the cloned cDNA yielded a 45 kDa protein. Human Aup1 coding region demonstrates 90% homology to its rat and mouse homologous at the nucleotide level, and 82% at the amine acid level. PMID- 12534238 TI - [A new human cellular protein AUP1. III. The intracellular localization of AUP1 protein in different human and rat cell lines]. AB - Previously, we cloned a full-length cDNA of human Aup1 and showed that AUP1 may represent a new cellular target for the two adenovirus oncoproteins, E1A Ad5 and E4ORF3. In this study, we generated a polyclonal anti-AUP1 antibody and examined the subcellular localization of AUP1 in MCF7 cells, HeLa cells, H1299 cells, 293 cells, BRK1 cells and transfectants expressing adenoviruse E1 genes. Double staining of AUP1 and various markers for cytoplasmic structures showed that the pattern of AUP1 distribution in the cytoplasm was puctuate and diffuse and without any colocalization with Golgi apparatus or endoplasmic reticulum. Additional studies with ectopically expressed AUP1, fused with red fluorescent protein (RFP) in H1299 and McG7 human cell lines and BRK1 rat cell line, showed cytoplasmic localization of RFP-AUP1. Western blot analysis revealed that AUP1 was expressed at similar levels in all tested cell lines and had the same molecular weight as the rat protein (45 kDa). Taken together, these results suggest that AUP1 is a cytoplasmic protein that is expressed in all cell lines we examined. PMID- 12534239 TI - [Impact of high-energy laser irradiation on pulmonic fibroblast in human embryo]. AB - This impact was attained by medicinal laser application to alexandrite, dye, erbium and gallium arsenid in various conditions on cultures of pulmonic fibroblasts of human embryo. Obtained results were estimated by transmission and scanning microscopy. The highest cell destruction was observed in dye and alexandrite, being less expressed in gallium arsenid under laser irradiation. Impulsive action of erbium laser did not cause any substantional cell destruction. So, laser application to dye alexandrite and gallium arsenide for healing scar and contracture damages is completely justified for skin pathogeny. PMID- 12534240 TI - The cellular basis of neurovascular metabolic coupling. AB - Functional brain imaging studies, such as positron emission tomography and functional nuclear magnetic resonance imaging, are based on the regional increases in cerebral blood flow, glucose consumption and oxygen consumption that are associated with regional increases in neuronal activity. The cellular basis of the signals that are measured is however incompletely understood at present. An increasingly important role for astrocytes in neurovascular metabolic coupling is being described, which was already suggested over 100 years ago as astrocytes are a structural link between endothelial cells of brain capillaries and neurons. Recent data have provided evidence for a role of the astrocytic intracellular sodium concentration in driving astrocytic glycolysis that may provide neurons with lactate as an energy substrate during activation. We have previously described intracellular sodium changes in astrocytes that are associated with astrocytic intercellular calcium waves. These intercellular calcium waves have been demonstrated both in astrocytes in culture as well as in brain slices. In this paper a new hypothesis concerning a role for astrocytic intercellular calcium waves in brain energy metabolism is formulated. PMID- 12534241 TI - The antiphospholipid syndrome. AB - The antiphospholipid syndrome (APS) defines the clinical association between antiphospholipid antibodies and a syndrome of hypercoagulability or thrombophilia (the term of "sticky blood" is sometimes used in APS). Antiphospholipid antibodies comprise a broad family of antibodies including both lupus anticoagulants and anticardiolipin antibodies. The pathogenesis of APS remains unclear. Nevertheless, an understanding of the biology, clinical and laboratory diagnosis, and clinical manifestations of APS are important to the neurologist because the brain is commonly affected by the disease. These points are addressed herein focusing on neurological manifestations of APS. Treatment of APS of which anticoagulation is the cornerstone is also discussed. PMID- 12534242 TI - Should levodopa be used anymore? AB - Levodopa is the most potent dopaminergic oral drug available in clinical practice. After chronic treatment, many patients with Parkinson's disease develop dyskinesia and motor fluctuations which are difficult to manage. It was hoped that introduction of dopaminergic agonists could diminish these side effects while keeping the same efficacy as levodopa. Prospective clinical data do not support this idea with the present drugs. Levodopa remains the most useful treatment and most clinicians believe that it is wise to associate early on levodopa with one of the dopamine agonists. PMID- 12534243 TI - Botulinum toxin-A therapy for palmar and plantar hyperhidrosis. AB - In an open study regarding focal hyperhidrosis, we injected 45-65 mouse units of botulinum toxin A (Btx-A) per palm and 100 per sole intracutaneously to 28 hands and 6 feet. We observed patients for up to 10 months to evaluate the efficacy and tolerability of Btx-A for palmar and plantar hyperhidrosis. The mean sweat production significantly declined for both palmar and plantar hyperhidrosis quantitatively on the first month of therapy (P < 0.01). One patient had transient muscle weakness and mild thenar atrophy interfering with her daily activities for 10 days. Injections were otherwise tolerated well by the patients. In this trial Btx-A injection is found to be an effective and safe method of treatment for palmar and plantar hyperhidrosis. PMID- 12534244 TI - Motor and phosphene thresholds to transcranial magnetic stimuli: a reproducibility study. AB - OBJECTIVES: As repetitive transcranial magnetic stimulation (rTMS) is often applied on different days, it is of interest to know whether motor (MT) and phosphene (PT) thresholds are reproducible across time and whether the intensity determined on the first day can be used in subsequent sessions. METHODS: We studied MT and PT over 5 separate recordings in 10 healthy volunteers using a focal coil and a Magstim(Rapid stimulator. After the initial recording (session 1), the others (2 to 5) were performed respectively after 1 day, 7 days, 1 month and 4 months. RESULTS: Mean MT at rest were 65.30 +/- 5.54%, 65.7 +/- 7.18%, 60.4 +/- 4.27%, 61.8 +/- 4.34%, and 63 +/- 9.1% at sessions 1 to 5. Mean PT were 71.43 +/- 6.68%, 66.29 +/- 10.67%, 60.71 +/- 8.64%, 60.57 +/- 8.08%, and 68.71 +/- 15.48% at sessions 1 to 5. MT and PT were reproducible (ANOVA analysis), however, as shown by coefficients of variation, variability between the first 3 sessions exceeded 10% for MT in 3 subjects and in 4 subjects for PT. CONCLUSIONS: It seems preferable to determine thresholds and adapt output intensity of the stimulator at each rTMS session. PMID- 12534245 TI - Brain function in the vegetative state. AB - Positron emission tomography (PET) techniques represent a useful tool to better understand the residual brain function in vegetative state patients. It has been shown that overall cerebral metabolic rates for glucose are massively reduced in this condition. However, the recovery of consciousness from vegetative state is not always associated with substantial changes in global metabolism. This finding led us to hypothesize that some vegetative patients are unconscious not just because of a global loss of neuronal function, but rather due to an altered activity in some critical brain regions and to the abolished functional connections between them. We used voxel-based Statistical Parametric Mapping (SPM) approaches to characterize the functional neuroanatomy of the vegetative state. The most dysfunctional brain regions were bilateral frontal and parieto temporal associative cortices. Despite the metabolic impairment, external stimulation still induced a significant neuronal activation (i.e., change in blood flow) in vegetative patients as shown by both auditory click stimuli and noxious somatosensory stimuli. However, this activation was limited to primary cortices and dissociated from higher-order associative cortices, thought to be necessary for conscious perception. Finally, we demonstrated that vegetative patients have impaired functional connections between distant cortical areas and between the thalami and the cortex and, more importantly, that recovery of consciousness is paralleled by a restoration of this cortico-thalamo-cortical interaction. PMID- 12534246 TI - More sleep cycles in the mid-luteal phase of a healthy control. PMID- 12534247 TI - A case of trigeminal-vagal neuralgia relieved by peripheral self-stimulation. AB - A case of simultaneous trigeminal and vagoglossopharyngeal neuralgia is described. No microvascular compression was seen at the trigeminal complex while the origin of vagoglossopharyngeal pain could have been due either to Chiari malformation or microvascular compression of the IX-X nerve complex. Decompressive surgery was ineffective. The patient could completely block his facial painful fits by strongly pinching the anterior axillary fold. This case militates against peripheral theories of facial neuralgias, including microvascular compression and ganglion ignition focus theories, and supports a central origin thereof. PMID- 12534248 TI - Biotransformation of alkaloids. AB - Biotransformations of alkaloids over the last decade have continued to encompass a wide variety of substrates and enzymes. The elucidation of novel alkaloid biosynthetic and catabolic pathways will continue to furnish new biocatalysts for the synthetic organic chemist. Furthermore, an improved understanding of the genetic and biochemical basis of metabolic pathways will also permit the engineering of pathways in plants and other heterologous hosts for the production of therapeutically important alkaloids. The combination of increasing commercial interest and advances in molecular biology will facilitate the availability of robust biocatalysts which are a prerequsite to achieve economically feasible processes for the production of alkaloid-based therapeutics. PMID- 12534249 TI - Putrescine, spermidine, spermine, and related polyamine alkaloids. PMID- 12534250 TI - Medicare cost controls and program compliance: the rationale of physician claims edits. AB - This article attempts to demystify and create a context for the enactment of several Medicare cost control and compliance systems for physician reimbursement. The focus is on claims "edits" and Medicare compliance. Portions of Medicare, including health care provider reimbursement, remain fee-for-service programs that can be easily defrauded. To protect the Trust, the Centers for Medicare and Medicaid Services (CMS) has taken a multi-pronged approach, using program administration, enforcement, and rules-based claims editing systems. The Evaluation and Management codes, the Correct Coding Initiative (CCI), and medical necessity rules are claims edits that affect procedure codes. The Medicare program has a complicated system of billing procedures and an apparatus to enforce them. A solid compliance plan must incorporate proper claims editing, because consistent incorrect Medicare billing can be considered abuse. Many resources are available to aid physicians, including computerized tools, new CMS initiatives, and Internet materials. PMID- 12534251 TI - Relative value units: from A to Z, Part 4. AB - The previous three articles in this series discussed relative value unit (RVU) basics and touched on some of the more practical applications of RVUs for managing a medical practice. 1-3 This fourth and final article illustrates the what, why, when, where and how of benchmarking RVUs. Benchmarking is often defined as a standard of measurement or evaluation, but in its truest sense, it is the ongoing process of establishing a standard of excellence and comparing activities to that standard. Health care has yet to establish true benchmarks, but it does have performance indicators for better-performing practices based on several criteria. PMID- 12534252 TI - Interviewing job applicants for a staff opening. AB - It can be very challenging to identify a potentially strong employee for your practice based upon a resume or written application alone. That is why a well structured and carefully prepared interview is such an important part of the recruitment process. However, many interviewers have vague interview goals or do not know how to make their limited time with the applicant count. This article describes the key ingredients for a successful interview and offers practical tips for interview preparation. Sample interview questions, legal considerations, and interview follow-up tips are offered, as well as practical advice about involving members of your staff in the interview process. PMID- 12534253 TI - Geriatric care managers: a collaborative resource to the physician practice. AB - Strongly independent and/or socially isolated elders with multiple health problems are often a physician's most challenging patient. These patients typically require intensive follow-up including phone calls from concerned long distance relatives, frequent and urgent same day physician visits and recurrent hospitalizations. This article will describe the role of a professional geriatric care manager (GCM). This article will discuss how a GCM can greatly assist both frail elders and physicians through comprehensive and ongoing assessment of an elder's needs, implementation of an optimal plan of care and close monitoring and management of information and supportive services. PMID- 12534254 TI - Physician mentoring: a process to maximize the success of new physicians and enhance synchronization of the group. AB - Of the various factors that serve as the ingredients for a healthy and successful medical group, none is more important than the people. In a medical group practice, physicians are the key to ensuring that the culture is grounded on a set of values and principles that promote teamwork and focus. Without it, the environment is destined to be grounded on hypocrisy and distrust. Organizations that take the time and commit the energy to effectively mentor new physicians will have the best chance of maintaining synchronization based on the alignment of individual expectation and the realities of the organization. PMID- 12534255 TI - Practice efficiency and physician compensation: a case study. AB - One of the major challenges facing today's health care executive is that of achieving maximum efficiency. The public also requires an efficient health care system. However, a problem occurs when the health care executive defines "efficiency" in a manner that is diametrically opposed to the public's definition. Maximum efficiency in the physician practice is defined by productivity equal to capacity. Maximum efficiency of the health care system is defined by lower health care costs. If the physician practice is achieving its goal of maximum efficiency, the market economy will likely force a failure in achieving the public's goal of a lower cost health care system! The following case study provides an opportunity to review this health care management conundrum and offers insights into a possible approach for some solution through attention to physician compensation. PMID- 12534256 TI - The Emergency Medical Treatment and Labor Act (EMTALA). AB - The Emergency Medical Treatment and Labor Act (EMTALA) was enacted to prevent patient dumping. It provides patients a considerable amount of protection and leverage in obtaining care under emergency situations. The reimbursement strictures imposed by managed care organizations have compounded the problems. This article summarizes the Act's definition of role and duties of hospitals and physicians and the regulations to which these parties must adhere or face significant penalties. PMID- 12534257 TI - Coding for injuries and skin grafts. PMID- 12534258 TI - Saving for college: 529 plans. PMID- 12534259 TI - First, do one thing right: deposit cash daily. PMID- 12534260 TI - Country living, city income for rural practitioners by implementing the Rural Health Clinics Act. AB - Rural medical practices have historically faced lower reimbursement than their urban counterparts. The relatively little-known Rural Health Clinics Act of 1976 permits a higher level of payment from Medicare and Medicaid through cost-based reimbursement. This article discusses the specifics and requirements of the Act and how practices can qualify and implement the change to rural health clinic participation. PMID- 12534261 TI - Improve compliance and financial performance at the same time. AB - Contrary to conventional wisdom, which holds that compliance is often a net negative to a practice's financial performance, the fact is that compliance, operations, and the financial performance of a medical practice can all be simultaneously improved. This article will illustrate that the basic drivers of effective compliance are often the same fundamental business principles that lead to outstanding operations and enhanced financial performance. The lesson for medical practice managers is that if you improve compliance, you should actually improve your bottom line, not harm it. PMID- 12534262 TI - Cutting out the middleman: physicians can contract directly with employers--a viable alternative to adversarial managed care agreements. AB - HMOs, PPOs, and other managed care "middlemen" control the means by which most physicians do business with employers. As physicians face dwindling reimbursements, greater practice restrictions, and increased pressure to sign adversarial middleman contracts, interest in direct contracting has grown. This article introduces direct contracting as an important alternative to commercial managed care agreements; cites the key advantages and process of direct contracting; and offers practical recommendations for helping physician practices successfully negotiate direct physician/employer agreements. PMID- 12534263 TI - [Factors of Yersinia pestis providing circulation and persistence of plague pathogen in ecosystems of natural foci. Communication 2]. AB - To maintain continuous circulation of plague pathogen in natural foci, the pathogen should be capable of invading host organism, resisting the bactericide protective systems of rodent, and reproducing itself to maintain the content of bacteria at a level sufficient for further transmission by fleas to a new host. Each of these stages of the Yersinia pestis circulation is determined by a variety of factors of plague pathogen, which may act either individually or in combination. Each of the factors itself may be involved in the pathological process at different stages of its development or in pathogen transmission. However, it is only the aggregate of the factors (regardless of significant or insignificant individual contribution to the sum effect) that provides persistence of plague pathogen in natural foci. The plague pathogen factors providing its transmission from one host organism to the next as well as correlation of individual factors of pathogensis and expression of various household genes with plague pathogensis virulence are considered in the second communication. This review was compiled on the basis of not only well-known works but also some sources of limited availability, particularly, for English-speaking audience. PMID- 12534264 TI - [Differences in virulence genes in Vibrio cholerae eltor strains isolated from different sources in Turkmenistan territory]. AB - Polymerase chain reaction (PCR) detected the presence of various genes associated with virulence in genome of strains V. cholerae eltor isolated in Turkmenistan territory during epidemic and epidemic-free perios. It was found that a complete set of virulence genes (ctxA+, tcpA+ and toxR+) contained strains isolated from patients, carriers and environment only in cholera epidemics. Strains isolated from the environment in the period free of epidemics did not contain ctxA and tcpA in 78.2% of cases, but 5.2% of the strains carried a complete set of virulence genes. There were also nontoxigenic strains containing genes tcpA and toxR. Such strains were isolated from the environment (16.6%) and vibrion carriers (42.9%). Isolated were also strains V.cholerae eltor carrying bacteriophage CTX phi with incomplete set of virulence genes and having genotype ctxA-, ace+ and zot+. Almost all the strains ctxA-, tcpA+ carry attRS1-site in genome. This shows that such strains may transform into toxigenic as a result of infection with bacteriophage CTX phi. PMID- 12534265 TI - [Genotyping of Yersinia pestis: variability of locus (CAAA)n in natural strains isolated in territory of former SSSR]. AB - Genome polymorphism by the locus (CAAA)n was studied in 69 strains of Yersinia pestis isolated from natural foci of the former Soviet Union. The polymorphism was found to be represented by ten alleles in chromosomes, which could be regarded as evidence of variability of this VNTR-locus (diversity index, DI = 0.86). The value of DI was found to vary substantially: from 0.24 in a group of vole strains from seven isolates from the Transcaucasian highlands to 0.77 in nine strains from the Central Asia desert focus. The allele polymorphism of the variable locus (CAAA)n in natural strains of Y. pestis was suggested to be used as a possible genetic marker of the strain. It was concluded that the oligonucleotide primers used in polymerase chain reaction should be upgraded to the genotyping accuracy. PMID- 12534266 TI - [Delivery of secreted placental alkaline phosphatase (SEAP) gene in vitro and in vivo as a component of recombinant avian adenovirus (CELO)]. AB - Recombinant adenoviruses capable of expressing the gene of secreted placentary alkaline phosphatase (SEAP) under control of CMV-promoter was obtained on the basis of CELO avian adenovirus and human adenovirus-5 (Ad5) genomes. The efficiency of the CELO vector was determined in experiments with transduction of human (293, A549, and H1299), mouse (B16), and avian (LMH) cell cultures. It was shown in C57BL/6 mice in vivo that SEAP gene is expressed under conditions of intravenous, intranasal, and intratumoral application of recombinant adenovirus CELO-SEAP. The duration of expression of the alkaline phosphatase CELO = SEAP gene in immunocompetent mouse body was 21 days. The level of SEAP gene expression was measured in the allantois fluid of chicken embryo infected with recombinant adenovirus CELO-SEAP. PMID- 12534267 TI - [Z-form of intraphage DNA]. AB - The bacteriophage lambda gt10 DNA containing an insertion of 20 pairs of GC-bases capable of being arranged as Z-form was cloned. Two independent methodological approaches based on the main properties of Z-form were used to study the shape of the insertion: formation of transition bridges composed of unpaired nucleotides between left-rotating Z-forms and right-rotating B-forms of helix (j-domain) and high immunogenic activity of Z-form. O-beta-diethylaminoethylhydroxyamine (OHA), an analogue of hydroxylamine, is capable of reacting specifically with unpaired cytidines. In this work this modification was used to inhibit the process of restriction at BamH1-site adjacent to (gc)10 insertion, that N-Methyl-bis(2 chloethyl) amine (MBCA) is capable of fixing the Z-form of the insertion in situ. Fixed Z-form is conserved even after DNA has been isolated from bacteriophage, thereby providing an opportunity of its identification by anti-Z-antibodies. It was shown that from 4 to 6% of the total number of insertions are in the Z-form. The hypothesis of significant role of Z-form in the process of packing of DNA molecules in capsid is put forward. PMID- 12534268 TI - [Genetic identification of the Crimean-Congo hemorrhagic fever virus during epidemic outbreak in Kazakhstan in 2000]. AB - Sera samples from patients suspected of Crimean-Congo hemorrhagic fever (CCHF) taken during epidemic outbreak at the territory of Sarysusky and Moiynkumsky districts of the Zhambyl region in Kazakhstan, in 2000, were analysed by means of reverse transcription-polymerase chain reaction (RT-PCR) and sequencing of virus genome fragments. Genome RNA of CCHF virus was found in 2 assays. Analysis of nucleotide sequences of fragments of S-segment of viral genome revealed in the Sarysusky districts circulation of CCHF virus, genetically resembled to close phylogenetically to CCHF virus strains from China. PMID- 12534269 TI - [Analysis of insertion-deletion polymorphism in serotonin transporter gene in men of different ethnic appurtenance with acute alcoholic psychosis]. AB - Analysis of insertion-deletion polymorphism of serotonin vector gene (SLC6A4) was carried out in Russian and Tatar men with acute alcoholic psychosis. Significant interpopulation differences in the distribution of SLC6A4 genotype and allele frequencies were revealed. A relationship of L/S gene with the disease was detected in Russians and Tatars, but the presence of heterozygotic genotype was associated with early onset of chronic alcoholization and development of acute alcoholic psychosis in Tatars and with later alcoholization and disease development in Russians. The share of S/S genotype was significantly decreased in Russian patients aged over 35 years, which suggests selection aimed at elimination of short allele homozygotes among patients with this disease and probably different genetic prerequisites for early and late development of the disease in Russians. In Tatars aged over 35 years acute alcoholic psychosis is associated with L/L genotype (RR-3). PMID- 12534270 TI - [Mode of sample preparation for pseudotuberculosis laboratory diagnosis by polymer chain reaction method]. AB - A mode of feces sample preparation was developed for polymerase chain reaction (PCR) assay. It was based on alkaline treatment of the material. This treatment killed the most part of indigenous microflora, whereas Yersinia survived, because it was relatively resistant to alkaline. The mode was tested using human feces artificially contaminated with Yersinia pseudotuberculosis. Positive responses in samples containing 10(3)-10(8) microbial cells per ml were obtained by PCR assay with Yersi and Yers2, Invl and Inv2, YP3 and YP4 primers. Diagnostic efficiency of PCR for patients, small mammals, and washings from environmental objects was 4.75, 1.66, and 2.12 times higher than diagnostic efficiency of bacteriological analysis of these samples, respectively. Positive results in PCR were obtained at the day of the material collection and treatment, whereas Y. pseudotuberculosis was isolated only after 8-20 days. Positive samples in PCR and in bacteriological analysis were found to coincide. A brief scheme of the Y. pseudotuberculosis laboratory diagnosis is suggested. According to this scheme, target-oriented bacteriological assay is performed only in those samples, in which preliminary PCR assay after 1-3 days of incubation gave positive results of Y. pseudotuberculosis DNA detection. PMID- 12534271 TI - Activation loop phosphorylation and catalysis in protein kinases: is there functional evidence for the autoinhibitor model? AB - Many protein kinases are activated strongly by the phosphorylation of a polypeptide region (activation loop) that lies outside the active-site cleft. Analysis of the X-ray crystallographic structures of the insulin receptor with the activation loop in the phosphorylated and dephosphorylated forms offers a testable model for the mechanism of activity regulation by the loop. In this model, the dephosphorylated activation loop can act as an autoinhibitor by blocking substrate access to the active site. Phosphorylation of the loop could then release the autoinhibitor from the active site, allowing substrate binding and catalysis. While this model has been widely invoked, it was not clear if solution studies would support an autoinhibitory model for kinase regulation, in general. We review the results of solution studies on six protein kinases that test the role of the activation loop in controlling active-site access. While loop phosphorylation enhances substrate binding in two cases, four protein kinases display little or no effect on substrate dissociation constants. By comparison, phosphorylation increases catalysis by 2-4 orders of magnitude in all cases. These findings can be used to place the phosphorylatable activation loops into two broad, functional subcategories. (i) Gated activation loops exhibit bifunctional properties restricting substrate access and controlling catalysis. (ii) Nongated activation loops allow free movement of the substrate in and out of the active site irrespective of phosphorylation state but potently modulate the phosphoryl transfer step. Thus, while activation loop phosphorylation greatly modulates catalytic potential, it does not necessarily affect substrate binding, as once widely believed. PMID- 12534272 TI - The structure of photosystem II in Arabidopsis: localization of the CP26 and CP29 antenna complexes. AB - A genetic approach has been adopted to investigate the organization of the light harvesting proteins in the photosystem II (PSII) complex in plants. PSII membrane fragments were prepared from wild-type Arabidopis thaliana and plants expressing antisense constructs to Lhcb4 and Lhcb5 genes, lacking CP29 and CP26, respectively (Andersson et al. (2001) Plant Cell 13, 1193-1204). Ordered PS II arrays and PS II supercomplexes were isolated from the membranes of plants lacking CP26 but could not be prepared from those lacking CP29. Membranes and supercomplexes lacking CP26 were less stable than those prepared from the wild type. Transmission electron microscopy aided by single-particle image analysis was applied to the ordered arrays and the isolated PSII complexes. The difference between the images obtained from wild type and antisense plants showed the location of CP26 to be near CP43 and one of the light-harvesting complex trimers. Therefore, the location of the CP26 within PSII was directly established for the first time, and the location of the CP29 complex was determined by elimination. Alterations in the packing of the PSII complexes in the thylakoid membrane also resulted from the absence of CP26. The minor light-harvesting complexes each have a unique location and important roles in the stabilization of the oligomeric PSII structure. PMID- 12534273 TI - The structure of the carboxyl terminus of striated alpha-tropomyosin in solution reveals an unusual parallel arrangement of interacting alpha-helices. AB - Coiled coils are well-known as oligomerization domains, but they are also important sites of protein-protein interactions. We determined the NMR solution structure and backbone (15)N relaxation rates of a disulfide cross-linked, two chain, 37-residue polypeptide containing the 34 C-terminal residues of striated muscle alpha-tropomyosin, TM9a(251-284). The peptide binds to the N-terminal region of TM and to the tropomyosin-binding domain of the regulatory protein, troponin T. Comparison of the NMR solution structure of TM9a(251-284) with the X ray structure of a related peptide [Li, Y., Mui, S., Brown, J. H., Strand, J., Reshetnikova, L., Tobacman, L. S., and Cohen, C. (2002) Proc. Natl. Acad. Sci. U.S.A. 99, 7378-7383] reveals significant differences. In solution, residues 253 269 (like most of the tropomyosin molecule) form a canonical coiled coil. Residues 270-279, however, are parallel, linear helices, novel for tropomyosin. The packing between the parallel helices results from unusual interface residues that are atypical for coiled coils. Y267 has poor packing at the coiled-coil interface and a lower R(2) relaxation rate than neighboring residues, suggesting there is conformational flexibility around this residue. The last five residues are nonhelical and flexible. The exposed surface presented by the parallel helices, and the flexibility around Y267 and the ends, may facilitate binding to troponin T and formation of complexes with the N-terminus of tropomyosin and actin. We propose that unusual packing and flexibility are general features of coiled-coil domains in proteins that are involved in intermolecular interactions. PMID- 12534274 TI - Structural and dynamic changes in human annexin VI induced by a phosphorylation mimicking mutation, T356D. AB - Phosphorylation of some members of the annexin family of proteins may play a significant role in controlling their calcium-dependent interactions with membranes. Recent electron microscopic studies of annexin VI revealed that the protein's two core domains exhibit a great degree of flexibility and are able to undergo a relative conformational change that could potentially initiate contacts between membranes [Avila-Sakar, A. J., et al. (2000) J. Struct. Biol. 130, 54 62]. To assess the possibility of a regulatory role of phosphorylation in this behavior, the crystal structure of a phosphorylation-mimicking mutant (T356D in the flexible connector region of human annexin VI) was determined to 2.65 A resolution. When the mutant is compared to the wild-type annexin VI, subtle differences are seen at the site of the mutation, while larger changes are evident in one of the calcium-binding loops and in the presence of five calcium ions. Furthermore, biochemical studies provide evidence for additional conformational differences between the T356D and wild-type solution structures. Fluorescence emission and acrylamide quenching suggest a higher level of solvent exposure of Trp-343 in the connector region of T356D in the presence of calcium. Comparisons of retardation coefficients in native gel electrophoresis reveal that T356D has a more extended shape, while proteolytic studies show a greater accessibility of a trypsin cleavage site inside the linker region, indicating a conformation more open than the wild-type form. These data provide insights into a possible regulatory mechanism leading to a higher degree of flexibility and possibly a higher calcium binding affinity of annexin VI upon phosphorylation. PMID- 12534275 TI - A major role for a set of non-active site mutations in the development of HIV-1 protease drug resistance. AB - A major problem in the chemotherapy of HIV-1 infection is the appearance of drug resistance. In the case of HIV-1 protease inhibitors, resistance originates from mutations in the protease molecule that lower the affinity of inhibitors while still maintaining a viable enzymatic profile. Drug resistance mutations can be classified as active site or non-active site mutations depending on their location within the protease molecule. Active site mutations directly affect drug/target interactions, and their action can be readily understood in structural terms. Non-active site mutations influence binding from distal locations, and their mechanism of action is not immediately apparent. In this paper, we have characterized a mutant form of the HIV-1 protease, ANAM-11, identified in clinical isolates from HIV-1 infected patients treated with protease inhibitors. This mutant protease contains 11 mutations, 10 of which are located outside the active site (L10I/M36I/S37D/M46I/R57K/L63P/A71V/G73S/L90M/I93L) and 1 within the active site (I84V). ANAM-11 lowers the binding affinity of indinavir, nelfinavir, saquinavir, and ritonavir by factors of 4000, 3300, 5800, and 80000, respectively. Surprisingly, most of the loss in inhibitor affinity is due to the non-active site mutations as demonstrated by additional experiments performed with a protease containing only the 10 non-active site mutations (NAM-10) and another containing only the active site mutation (A-1). Kinetic analysis with two different substrates yielded comparable catalytic efficiencies for A-1, ANAM-11, NAM-10, and the wild-type protease. These studies demonstrate that non-active site mutations can be the primary source of resistance and that their role is not necessarily limited to compensate deleterious effects of active site mutations. Analysis of the structural stability of the proteases by differential scanning calorimetry reveals that ANAM-11 and NAM-10 are structurally more stable than the wild-type protease while A-1 is less stable. Together, the binding and structural thermodynamic results suggest that the non-active site mutants affect inhibitor binding by altering the geometry of the binding site cavity through the accumulation of mutations within the core of the protease molecule. PMID- 12534276 TI - Solution structure of the RNase H domain of the HIV-1 reverse transcriptase in the presence of magnesium. AB - This paper presents the first solution structure of the RNase H domain of HIV-1 reverse transcriptase (RT) determined by NMR methods. The solution conditions in this study were at physiological pH in the presence of Mg(2+). An investigation of the dependence of the (1)H-(15)N HSQC spectrum of the RNase H domain on [Mg(2+)] indicates that Mg(2+) produces significant, global effects on the amide chemical shifts, implying that divalent metal ion binding is important for stabilizing the structure of the isolated domain in solution. Analysis of amide shift data as a function of MgCl(2) concentration using either a single- or two site binding model indicated that the latter provided a significantly improved fit, with the K(D) for site A = 2.7-3.2 mM and K(D) for site B approximately 35 mM, calculated on the assumption that site A is already occupied. Resonances of the [U-(13)C,(15)N]RNase H domain, measured at pH 6.8, in 80 mM MgCl(2), were assigned and NOESY data collected in order to determine the structure. Assignment of the NOESY spectra using the ARIA program resulted in a high-resolution structure for residues 6-114 which was similar to the crystal structure of the isolated domain,. The data were insufficient to define a compact structure for the C-terminal residues after 114. Residues I134-L138 located at the C-terminus are highly disordered and give rise to relatively sharp and intense amide resonances, while the amide resonances for the segment from E124 to A132 appear to be largely absent and are presumably subject to significant exchange broadening between different conformational states. Comparisons with crystal structure data for the full reverse transcriptase molecule indicate that the corresponding region is absent in nearly all of the crystal structures determined for the P2(1)2(1)2(1) space group, while these residues adopt an alpha-helix in structures determined for other symmetry groups. This structural heterogeneity indicates that significant conformational variability exists for this segment of the full reverse transcriptase enzyme as well, and the structure of the C terminal peptide can be selected or deselected, depending on crystallization conditions. This analysis, along with the structural characterization contained herein, challenges the previous paradigm that the dynamic behavior of the isolated RNase H domain differs substantially from the behavior in the intact enzyme. The poor Mg(2+) binding and conformational flexibility of residues located near the active site indicate that substrate binding is a precondition for metal ion binding and for selecting the active site conformation of the RNase H domain. PMID- 12534277 TI - NMR study of the interaction between Zn(II) ligated bleomycin and Streptoalloteichus hindustanus bleomycin resistance protein. AB - Bleomycin (Bm), a 1.4 kDa glycopeptide excreted by Streptomyces verticillus, is a natural antibacterial compound used in therapy as antineoplastic drug. To counteract its biological activity, cells have developed several resistance mechanisms, one of these based on proteins able to tightly bind Bm. In this paper, the interaction of Zn(2+)-Bm with the Streptoalloteichus hindustanus Bm resistance protein (ShBle) has been investigated by solution state NMR. Sequential nOe and chemical shift index have shown that the fold of the protein (in absence or presence of Bm) is identical to the previously published X-ray structure. The dimeric nature of ShBle is confirmed by the diffusion tensor as determined by NMR relaxation data. Using isotope filtered nOe experiment, intermolecular nOes between Bm and ShBle have been observed as used for modeling. While the interaction of the Bm metal binding site with ShBle appears to be uniquely defined, several conformations of the bithiazole moieties are compatible with the NMR data. Binding of Bm also induces changes of the local dynamics (stretch N85-G91), as shown by (15)N relaxation data. These results are discussed in the context of several Bm analogues able to interact with ShBle and of the recently published X-rays structures. PMID- 12534278 TI - Large-scale context in protein folding: villin headpiece. AB - The villin headpiece folds autonomously in vitro forming three alpha-helical regions. Local propensities, however, strongly disfavor the formation of the C terminal helix because most native residue pairs in that helix are hydrophobic/polar mismatches. Even the N-terminal helix is unfavored according to the AGADIR criterion. Our coarse-grained ab initio simulations reveal three-body correlations in which hydrophobic residues position to protect amide-carbonyl hydrogen bonds from attack by water, thus inducing the growth of the C-terminal helix and guiding the folding process. Similar correlations are also found in all atom simulations with an implicit solvent model that accurately reproduces the results of simulations with explicit solvent molecules. The correlations establish a large-scale, many-body context that may be probed experimentally by introducing mutations of certain nonobvious residues that reside outside the native hydrophobic core but that are predicted to affect the folding rates and dynamics dramatically. PMID- 12534279 TI - The N-terminal repeat domain of alpha-synuclein inhibits beta-sheet and amyloid fibril formation. AB - The conversion of alpha-synuclein into amyloid fibrils in the substantia nigra is linked to Parkinson's disease. Alpha-synuclein is natively unfolded in solution, but can be induced to form either alpha-helical or beta-sheet structure depending on its concentration and the solution conditions. The N-terminus of alpha synuclein comprises seven 11-amino acid repeats (XKTKEGVXXXX) which can form an amphipathic alpha-helix. Why seven repeats, rather than six or eight, survived the evolutionary process is not clear. To probe this question, two sequence variants of alpha-synuclein, one with two fewer (del2) and one with two additional (plus2) repeats, were studied. As compared to wild-type alpha synuclein, the plus2 variant disfavors the formation of beta-sheet-rich oligomers, including amyloid fibrils. In contrast, the truncated variant, del2, favors beta-sheet and fibril formation. We propose that the repeat number in WT alpha-synuclein represents an evolutionary balance between the functional conformer of alpha-synuclein (alpha-helix and/or random coil) and its pathogenic beta-sheet conformation. N-terminal truncation of alpha-synuclein may promote pathogenesis. PMID- 12534280 TI - Assignment of heme resonances and determination of the electronic structures of high- and low-spin nitrophorin 2 by 1H and 13C NMR spectroscopy: an explanation of the order of heme methyl resonances in high-spin ferriheme proteins. AB - The (1)H NMR resonances of the heme substituents of the low-spin Fe(III) form of nitrophorin 2, as its complexes with N-methylimidazole (NP2-NMeIm) and imidazole (NP2-ImH), have been assigned by a combination of (1)H homonuclear two dimensional NMR techniques and (1)H-(13)C HMQC. Complete assignment of the proton and partial assignment of the (13)C resonances of the heme of these complexes has been achieved. Due to favorable rates of ligand exchange, it was also possible to assign part of the (1)H resonances of the high-spin heme via saturation transfer between high- and low-spin protein forms in a partially liganded NP2-NMeIm sample; additional resonances (vinyl and propionate) were assigned by NOESY techniques. The order of heme methyl resonances in the high-spin form of the protein over the temperature range of 10-37 degrees C is 8 = 5 > 1 > 3; the NMeIm complex has 5 > 1 > 3 > 8 as the order of heme methyl resonances at <30 degrees C, while above that temperature, the order is 5 > 3 > 1 > 8, due to crossover of the closely spaced 3- and 1-methyl resonances of the low-spin complex at higher temperatures. This crossover defines the nodal plane of the heme orbital used for spin delocalization as being oriented 162 +/- 2 degrees clockwise from the heme N(II)-Fe-N(IV) axis for the heme in the B orientation. For the NP2-ImH complex, the order of heme methyl resonances is 3 > 5 > 1 > 8, which defines the orientation of the nodal plane of the heme orbital used for spin delocalization as being oriented approximately 150-155 degrees clockwise from the heme N(II)-Fe N(IV) axis. In both low-spin complexes, the results are most consistent with the exogenous planar ligand controlling the orientation of the nodal plane of the heme orbital. In the high-spin form of NP2, the proximal histidine plane is shown to be oriented 135 degrees clockwise from the heme N(II)-Fe-N(IV) axis, again for the B heme orientation. A correlation between the order of heme methyl resonances in the high-spin form of NP2 and several other ferriheme proteins and an apparent 90 degrees shift in the nodal plane of the orbital involved in spin delocalization from that expected on the basis of the orientation of the axial histidine imidazole nodal plane have been explained in terms of bonding interactions between Fe(III), the axial histidine imidazole nitrogen, and the porphyrin pi orbitals of the high-spin protein. PMID- 12534281 TI - Structural requirements for catalysis, expression, and dimerization in the CD26/DPIV gene family. AB - Dipeptidyl peptidase IV (DP-IV/CD26), fibroblast activation protein (FAP), DP like 1 (DPL1), DP8, DP9, and DPL2 comprise the CD26 gene family. CD26/DP-IV has roles in liver disease, T cell costimulation, chemokine biology, type II diabetes, and tumor biology. DPIV substrates include the glucagonlike peptides, neuropeptide Y, and the chemokines CCL3, CCL5, CCL11, CCL22, and CXCL12. We have proposed that the extracellular region of CD26 is analogous to prolyl oligopeptidase in consisting of an alpha/beta hydrolase domain contributed by both N- and C-terminal portions of the polypeptide and a seven-blade beta propeller domain. Replacing the C-terminal portion of the predicted alpha/beta hydrolase domain of CD26 (residues 501-766) with the homologous portion of DP8 or DP9 produced intact proteins. However, these chimeric proteins lacked dimerization and peptidase activity, suggesting that CD26 dimerization requires the C-terminal portion of the alpha/beta hydrolase domain. Deleting some N terminal residues of the alpha/beta hydrolase domain of CD26 ablated peptidase activity and greatly diminished cell surface expression. Together with previous data that CD26 peptidase activity requires the C-terminal 20 residues, this suggests that peptidase activity requires the entire alpha/beta hydrolase domain. The catalytic triad of DP8 was shown to be Ser(739)-Asp (817)-His(849). Glu(259) of DP8, a residue distant from the catalytic triad yet greatly conserved in the CD26 gene family, was shown to be required for peptidase activity. These data concord with our predicted CD26 structure, indicate that biosynthesis of a functional fragment of CD26 is difficult, and confirm the functional homology of DP8 with CD26. PMID- 12534282 TI - Protease-activated receptor mediated RhoA signaling and cytoskeletal reorganization in LNCaP cells. AB - Thrombin and trypsin induce cell signaling through a subclass of G-protein coupled receptors called the protease-activated receptors (PARs). In many cells, PAR signaling results in the activation of RhoA and other members of the Rho family of small GTPases which are involved in cytoskeletal reorganization. The expression of PARs and their role in the activation of Rho GTPases in prostate cancer cells are not clearly known. FACS analysis demonstrated that the androgen dependent LNCaP cells express PAR1, PAR2, and PAR4 but not PAR3. Stimulation with thrombin and trypsin resulted in the rapid activation of RhoA in a dose-dependent manner with an EC(50) of 1.0 and 5 nM, respectively. Activation of RhoA was enhanced by, but not dependent on, the presence of 1 nM dihydrotestosterone. Inhibition of the proteolytic properties of thrombin by hirudin and trypsin by diisopropyl fluorophosphate abolished the observed RhoA activation. Stimulation with 150 microM PAR-activating peptides TFFLRN (PAR1), SLIGKV (PAR2), and AYPGKF (PAR4) demonstrated that PAR1 and PAR2 mediated protease-activated RhoA signaling. Fluorescent microscopy studies showed that LNCaP cells treated with either thrombin (10 nM) or trypsin (10 nM) developed an increased number of filopodia, stress fibers, and focal adhesions relative to untreated cells. These observations represent the first report of PAR signaling in prostate cancer cells as well as the ability of PAR2 to mediate RhoA activation. Since the activation of RhoA is important for cytoskeletal reorganization, we postulate that PAR mediated RhoA activation may be a major signaling pathway in the biology of prostate cancer. PMID- 12534283 TI - Metal ion enhanced binding of AMD3100 to Asp262 in the CXCR4 receptor. AB - The affinity of AMD3100, a symmetrical nonpeptide antagonist composed of two 1,4,8,11-tetraazacyclotetradecane (cyclam) rings connected through a 1,4 dimethylene(phenylene) linker to the CXCR4 chemokine receptor was increased 7, 36, and 50-fold, respectively, by incorporation of the following: Cu(2+), Zn(2+), or Ni(2+) into the cyclam rings of the compound. The rank order of the transition metal ions correlated with the calculated binding energy between free acetate and the metal ions coordinated in a cyclam ring. Construction of AMD3100 substituted with only a single Cu(2+) or Ni(2+) ion demonstrated that the increase in binding affinity of the metal ion substituted bicyclam is achieved through an enhanced interaction of just one of the ring systems. Mutational analysis of potential metal ion binding residues in the main ligand binding crevice of the CXCR4 receptor showed that although binding of the bicyclam is dependent on both Asp(171) and Asp(262), the enhancing effect of the metal ion was selectively eliminated by substitution of Asp(262) located at the extracellular end of TM-VI. It is concluded that the increased binding affinity of the metal ion substituted AMD3100 is obtained through enhanced interaction of one of the cyclam ring systems with the carboxylate group of Asp(262). It is suggested that this occurs through a strong concomitant interaction of one of the oxygen's directly with the metal ion and the other oxygen to one of the nitrogens of the cyclam ring through a hydrogen bond. PMID- 12534284 TI - Influenza neuraminidase inhibitors: structure-based design of a novel inhibitor series. AB - Combinatorial and structure-based medicinal chemistry strategies were used together to advance a lead compound with an activity of K(i) = 58 microM via a potency enhancement of >70 000-fold to an analogue with an activity of K(i) = 0.8 nM against influenza neuraminidase (A/Tokyo/67). Lead optimization was initiated using molecular modeling and combinatorial chemistry. Protein crystal structures revealed that inconsistent structure-activity relationship (SAR) data resulted from different binding orientations of the inhibitor core five-membered rings from one series to another. Binding modes for a series of compounds showed up to a 180 degrees variation in orientation of the five-membered ring within the active site. Potent analogues were only achieved with chemical series that were observed to bind in the same orientation and yielded consistent SAR. In one series, consistent binding was obtained by an unprecedented occupation of a negatively charged binding pocket by a neutral methyl ester unit. The structural rationale for this novel SAR variation, based on protein crystallographic data, is given. PMID- 12534285 TI - Light affects the accessibility of the thylakoid light harvesting complex II (LHCII) phosphorylation site to the membrane protein kinase(s). AB - Redox-controlled, reversible phosphorylation of the thylakoid light harvesting complex II (LHCII) regulates its association with photosystems (PS) I or II and thus, energy distribution between the two photosystems (state transition). Illumination of solubilized LHCII enhances exposure of the phosphorylation site at its N-terminal domain to protein kinase(s) and tryptic cleavage in vitro [Zer et al. (1999) Proc. Natl. Acad. Sci. U.S.A. 96, 8277-8282]. Here we report that short illumination (5-10 min, 15-30 micromol m(-2) s(-1)) enhances the accessibility of LHCII phosphorylation site to kinase(s) activity also in isolated thylakoids. However, prolonged illumination or higher light intensities (30 min, 80-800 micromol m(-2) s(-1)) prevent phosphorylation of LHCII in the isolated membranes as well as in vivo, although redox-dependent protein kinase activity persists in the illuminated thylakoids toward exogenous solubilized LHCII. This phenomenon, ascribed to light-induced inaccessibility of the phosphorylation site to the protein kinase(s), affects in a similar way the accessibility of thylakoid LHCII N-terminal domain to tryptic cleavage. The illumination effect is not redox related, decreases linearly with temperature from 25 to 5 degrees C and may be ascribed to light-induced conformational changes in the complex causing lateral aggregation of dephosphorylated LHCII bound to and/or dissociated from PSII. The later state occurs under conditions allowing turnover of the phospho-LHCII phosphate. The light-induced inaccessibility of LHCII to the membrane-bound protein kinase reverses readily in darkness only if induced under LHCII-phosphate turnover conditions. Thus, phosphorylation prevents irreversible light-induced conformational changes in LHCII allowing lateral migration of the complex and the related state transition process. PMID- 12534286 TI - Structure and dynamics of reduced Bacillus pasteurii cytochrome c: oxidation state dependent properties and implications for electron transfer processes. AB - The solution structure of reduced Bacillus pasteurii cytochrome c, which has only 71 amino acids, has been determined by NMR to an RMSD of 0.46 +/- 0.08 A for all backbone atoms and 0.79 +/- 0.08 A for all heavy atoms and refined through restrained energy minimization. The target function out of 1645 constraints is 0.52 +/- 0.11 A(2), and the penalty function is 66 +/- 12 kJ mol(-)(1). The structure appears very similar to that in the oxidized state, only Trp87 and the propionates showing significant differences. The mobility was investigated through (15)N R(1) and R(2) relaxation rates, (15)N-(1)H NOE, and (1)H/(2)H exchange. It is found that the oxidized form is generally more mobile than the reduced one. By comparing the redox-state dependence of the structural/dynamic properties of Fe-S proteins, cytochrome c, and blue copper proteins, hints are provided for a better comprehension of the electron transfer processes. PMID- 12534287 TI - The ND5 subunit was labeled by a photoaffinity analogue of fenpyroximate in bovine mitochondrial complex I. AB - Fenpyroximate is a potent inhibitor of the mitochondrial proton-translocating NADH-quinone oxidoreductase (complex I). We synthesized its photoaffinity analogue [(3)H](trifluoromethyl)phenyldiazirinylfenpyroximate ([(3)H]TDF). When bovine heart submitochondrial particles (SMP) were illuminated with UV light in the presence of [(3)H]TDF, radioactivity was mostly incorporated into a 50 kDa band. There was a good correlation between radioactivity labeling of the 50 kDa band and inhibition of the NADH oxidase activity, indicating that a 50 kDa protein is responsible for the inactivation of complex I. Blue native gel electrophoresis of the [(3)H]TDF-labeled SMP revealed that the majority of radioactivity was found in complex I. Analysis of the complex I band on an SDS gel showed a major peak of radioactivity at approximately 50 kDa. There are three subunits in complex I that migrate in this region: FP51K, IP49K, and ND5. Further analysis using the 2D gel electrophoresis implied that the labeled protein was the ND5 subunit. Labeling of the ND5 subunit was stimulated by NADH/NADPH but was prevented by various complex I inhibitors. Amiloride derivatives that are known to be inhibitors of Na(+)/H(+) antiporters also diminished the labeling. In agreement with the protective effect, we observed that the amiloride derivatives inhibited NADH-ubiquinone-1 reductase activity but not NADH-K(3)Fe(CN)(6) reductase activity in bovine SMP. These results suggest that the ND5 subunit is involved in construction of the inhibitor- and quinone-binding site(s). Furthermore, it seems likely that the ND5 subunit may participate in H(+)(Na(+)) translocation in coupling site 1. PMID- 12534288 TI - Identification of membrane topography of the electrogenic sodium bicarbonate cotransporter pNBC1 by in vitro transcription/translation. AB - The transmembrane topography of the human pancreatic electrogenic sodium bicarbonate cotransporter pNBC1 was investigated using in vitro transcription/translation of HK-M0 and HK-M1 fusion vectors designed to test membrane insertion properties of pNBC1 hydrophobic sequences (H). These vectors encode N-terminal 101 (HK-M0) or 139 (HK-M1) amino acids of the H,K-ATPase alpha subunit, a linker region and the C-terminal 177 amino acids of the H,K-ATPase beta-subunit that contain five N-linked glycosylation consensus sites (Bamberg, K., and Sachs, G. (1994) J. Biol. Chem. 269, 16909-16919). The glycosylation status of the beta-region was used as a reporter to determine whether a given hydrophobic sequence possesses signal anchor and/or stop transfer properties in the HK-M0 and HK-M1 vectors. The linker region of each vector was replaced either with individual hydrophobic sequences or combinations thereof. The transcription/translation products of these fusion vectors in reticulocyte lysate system +/- microsomal membranes were identified by [(35)S]-autoradiography following separation using SDS-PAGE. The results of the in vitro transcription/translation analysis indicated that 10 (H1, H2N, H3, H5, H6, H7, H8, H9, H11, and H12) out of 12 hydrophobic sequences were able to insert into the plasma membrane. Two hydrophobic sequences, H4 and H10, had no membrane insertion activity even when upstream and downstream sequences were present. These data and immunocytochemical studies indicate that pNBC1 contains 10 transmembrane domains with N- and C-termini oriented intracellularly. This is the first characterization of the membrane topography of a sodium bicarbonate cotransporter. PMID- 12534290 TI - Differential recognition of the free versus bound retinol by human microsomal retinol/sterol dehydrogenases: characterization of the holo-CRBP dehydrogenase activity of RoDH-4. AB - All-trans-retinol is the precursor for all-trans-retinoic acid, the activating ligand for nuclear transcription factors retinoic acid receptors. In the cytosol of various cells, most retinol exists in a bound form, complexed with cellular retinol binding protein type I (holo-CRBP). Whether retinoic acid is produced from the free or bound form of retinol is not yet clear. Here, we present evidence that holo-CRBP is recognized as substrate by human microsomal short chain dehydrogenase/reductase (SDR) RoDH-4 with the K(m) value close to the liver concentration of holo-CRBP. The ability to utilize holo-CRBP differentiates RoDH 4 from a related enzyme, RoDH-like 3alpha-hydroxysteroid dehydrogenase (3alpha HSD), which is 3-fold more active with free retinol than RoDH-4 but is 15-fold less active toward holo-CRBP. Recognition of the cytosolic holo-CRBP as substrate is consistent with RoDH-4 orientation in the membrane. As established by immunoprecipitation and glycosylation scanning, RoDH-4 faces the cytosolic side of the membrane. Purified RoDH-4, stabilized by reconstitution into proteoliposomes, exhibits the apparent K(m) values for substrates and NAD(+) similar to those of the microsomal enzyme and oxidizes holo-CRBP with the catalytic efficiency (k(cat)/K(m)) of 59 min(-1) mM(-1). Apo-CRBP acts as a strong competitive inhibitor of holo-CRBP oxidation with an apparent K(i) value of 0.2 microM. The results of this study suggest that the human retinol-active SDRs are not functionally equivalent and that, in contrast to RoDH-like 3alpha HSD, RoDH-4 can access the bound form of retinol for retinoic acid production and is regulated by the apo-/holo-CRBP ratio. PMID- 12534289 TI - Dimeric structure of the six-domain VibF subunit of vibriobactin synthetase: mutant domain activity regain and ultracentrifugation studies. AB - Nonribosomal peptide synthetases (NRPS), fatty acid synthases (FAS), and polyketide sythases (PKS) are multimodular enzymatic assembly lines utilized in natural product biosynthesis. Previous data on FAS and PKS subunits have indicated that they are homodimers and that some of their catalytic functions can work in trans. When NRPS assembly lines have been probed for comparable formation of stable oligomers, no evidence had been forthcoming that species other than monomer forms were active. In this work we focus on the six-domain (Cy1-Cy2-A-C1 PCP-C2) enzyme VibF from the vibriobactin synthetase assembly line, which contains three other proteins, VibB, VibE, and VibH, that--when purified and mixed with VibF and the substrates ATP, threonine, 2,3-dihydroxybenzoate (DHB), and norspermidine--produce the iron chelator vibriobactin. Using a deletion of the Cy1 domain and separate inactivating mutations in the Cy2, A, PCP, and C2 domains of VibF, we report regain of catalytic activity upon mutant protein mixing that argues for heterodimer formation, stable for hundreds to thousands of catalytic cycles, with acyl chain processing and transfer around blocked domains. Ultracentrifugation data likewise confirm a dimeric structure for VibF and establish that domains within NRPS dimeric modules can act on acyl chains in trans. The results described here are the first indication for an NRPS subunit that homodimerization can occur and that there is a continuum of functional oligomerization states between monomers and dimers in nonribosomal peptide synthetases. PMID- 12534291 TI - Metal ion binding and enzymatic mechanism of Methanococcus jannaschii RNase HII. AB - RNase H degrades the RNA moiety in DNA:RNA hybrid in a divalent metal ion dependent manner. It is essential to understand the role of metal ion in enzymatic mechanism. One of the key points in this study is how many metal ions are involved in the enzyme catalysis. Accordingly, either one-metal binding mechanism or two-metal binding mechanism is proposed. We have studied the thermodynamic properties of four metal ions (Mg(2+), Mn(2+), Ca(2+), and Ba(2+)) binding to Methanococcus jannaschii RNase HII using isothermal titration calorimetry. All of the four metal ions were found to bind Mj RNase HII with 1:1 stoichiometry in the absence of substrate. Together with enzymatic activity assay data, we propose that only one metal ion binding to the enzyme in catalytic process. We also studied the pH dependence of metal binding and enzyme activity and found that at pH 6.5, Mg(2+) did not bind to the enzyme without the substrate but still activated the enzyme to about 2% of its maximum activity (in 10 mM Mn(2+) at pH 8). This implies that the substrate may also be incorporated in metal ion binding and help to position the metal ion. To find which acidic residues correspond to metal ion binding, we also studied the binding thermodynamics and enzymatic activity assay of four mutants: D7N, E8Q, D112N, and D149N in the presence of Mn(2+). The thermodynamic parameters are least affected for the D149N mutant, which has a very low enzymatic activity. This indicates that Asp149 is essential for the enzymatic activity. On the basis of all these observations, we suggest a metal binding model in which D7, E8, and D112 bind the metal ion and D149 activates a water molecule to attack the P-O bond in the RNA chain of the substrate. PMID- 12534292 TI - DNA binding by antitumor trans-[PtCl2(NH3)(thiazole)]. Protein recognition and nucleotide excision repair of monofunctional adducts. AB - Antitumor effects of cis-diamminedichloroplatinum(II) (cisplatin) and the clinical inactivity of its trans isomer (transplatin) have been considered a paradigm for the classical structure-activity relationships of platinum drugs. However, several new analogues of transplatin which exhibit a different spectrum of cytostatic activity including activity in tumor cells resistant to cisplatin have been recently identified. Analogues containing the planar amine ligand of the general structure trans-[PtCl(2)(NH(3))(L)], where L = planar amine, represent an example of such compounds. DNA is believed to be the major pharmacological target of platinum compounds. To contribute to the understanding of mechanisms underlying the activation of trans geometry in transplatin analogues containing planar amine ligands, various biochemical and biophysical methods were employed in previous studies to analyze the global modifications of natural DNA by trans-[PtCl(2)(NH(3))(L)]. These initial studies have revealed some unique features of the DNA binding mode of this class of platinum drugs. As the monofunctional lesions represent a significant fraction of stable adducts formed in DNA by bifunctional antitumor trans-platinum compounds with planar ligands, we analyzed in the present work short DNA duplexes containing the single, site-specific monofunctional adduct of a representative of this class of platinum drugs, antitumor trans-[PtCl(2)(NH(3))(thiazole)]. It has been shown that, in contrast to the adducts of monodentate chlorodiethylenetriamineplatinum(II) chloride or [PtCl(NH(3))(3)]Cl, the monofunctional adduct of trans-[PtCl(2)(NH(3))(thiazole)] inhibits DNA synthesis and creates a local conformational distortion similar to that produced in DNA by the major 1,2-GG intrastrand CL of cisplatin, which is considered the lesion most responsible for its anticancer activity. In addition, the monofunctional adducts of trans-[PtCl(2)(NH(3))(thiazole)] are recognized by HMGB1 domain proteins and removed by the nucleotide excision repair system similarly as the 1,2-GG intrastrand CL of cisplatin. The results of the present work further support the view that the simple chemical modification of the structure of an inactive platinum compound alters its DNA binding mode into that of an active drug and that processing of the monofunctional DNA adducts of the trans-platinum analogues in tumor cells may be similar to that of the major bifunctional adducts of "classical" cisplatin. PMID- 12534293 TI - Escherichia coli MutY and Fpg utilize a processive mechanism for target location. AB - MutY and formamidopyrimidine-DNA-glycosylase (Fpg) are base-excision repair (BER) enzymes involved in the 8-oxoguanine repair pathway in Escherichia coli. An impressive feature of these enzymes is the ability to locate 8-oxoguanine lesions among a large excess of undamaged DNA. To provide insight into the mechanism of target location, the ability of these enzyme to utilize a one-dimensional processive search (DNA sliding) or distributive (random diffusion-mediated) mechanism was investigated. Each enzyme was incubated with double-stranded concatemeric polynucleotides containing a site-specific target site at 25 nucleotide (nt) intervals. The products of each reaction were analyzed after further treatment and denaturation. A rapid accumulation of predominantly 25-nt fragments would indicate the utilization of a processive mechanism, whereas oligomeric multiples of 25-nt fragments would form if a distributive mechanism were used. Both Fpg and MutY were found to function processively on concatemers containing 7,8-dihydro-8-oxo-2'-deoxyguanosine (OG).C and G.A mispairs, respectively. An increase in sodium chloride concentration results in the modulation from a processive to distributive mechanism for both enzymes. Interestingly, processive behavior was not observed in the reaction of MutY with concatemers containing OG.A mispairs. A truncated form of MutY (Stop 225) containing only the N-terminal domain was found to behave in a manner consistent with a processive mechanism with both OG.A- and G.A-containing substrates. This suggests that the C-terminal domain of MutY plays an important role in the mechanism by which the enzyme detects OG.A base pairs in DNA. PMID- 12534294 TI - Phosphorylation of RGS14 by protein kinase A potentiates its activity toward G alpha i. AB - Regulators of G protein signaling (RGS proteins) modulate Galpha-directed signals because of the GTPase activating protein (GAP) activity of their conserved RGS domain. RGS14 and RGS12 are unique among RGS proteins in that they also regulate Galpha(i) signals because of the guanine nucleotide dissociation inhibitor (GDI) activity of a GoLoco motif near their carboxy-termini. Little is known about cellular regulation of RGS proteins, although several are phosphorylated in response to G-protein directed signals. Here we show for the first time the phosphorylation of native and recombinant RGS14 in host cells. Direct stimulation of adenylyl cyclase or introduction of dibutyryl-cAMP induces phosphorylation of RGS14 in cells. This phosphorylation occurs through activation of cAMP-dependent protein kinase (PKA) since phosphate incorporation is completely blocked by a selective inhibitor of PKA but only partially or not at all blocked by inhibitors of other G-protein regulated kinases. We show that purified PKA phosphorylates two specific sites on recombinant RGS14, one of which, threonine 494 (Thr494), is immediately adjacent to the GoLoco motif. Because of this proximity, we focused on the possible effects of PKA phosphorylation on the GDI activity of RGS14. We found that mimicking phosphorylation on Thr494 enhanced the GDI activity of RGS14 toward Galpha(i) nearly 3-fold, with no associated effect on the GAP activity toward either Galpha(i) or Galpha(o). These findings implicate cAMP-induced phosphorylation as an important modulator of RGS14 function since phosphorylation could enhance RGS14 binding to Galpha(i)-GDP, thereby limiting Galpha(i) interactions with downstream effector(s) and/or enhancing Gbetagamma-dependent signals. PMID- 12534295 TI - Assessment of membrane-bound mammal mitochondrial adenine nucleotide translocase topography by experimental antibodies. AB - To gain insight into the immunogenicity of mitochondrial adenine nucleotide translocase (ANT), we raised antibodies against purified bovine heart ANT by induction of ascitic fluid in male Balb/c mice. We identified the antigenic determinants detected by these antibodies by (1) immunodetection of GST-ANT fusion proteins and selected partial constructs of ANT, (2) immunodetection of chemically synthesized overlapping peptides on solid support, and (3) back titration ELISA. Results revealed a short epitope spreading of the antibodies, resulting in a small number of antigenic determinants. Thus, each antibody detects one or two major epitopes located in the putative hydrophilic loops M2 and M3. No evidence for the antigenicity of the first 133 amino acids of ANT was obtained. These well-characterized antibodies were used to study the topography of the membrane-bound ANT by back-titration ELISA with mitochondrial membranes. We demonstrated that amino acids 145-150 and 230-237 are fully accessible to the antibodies in native ANT, whereas regions 133-140 and 244-251 are not. Furthermore, we used mitochondria devoid of the outer membrane (mitoplasts) and inside-out submitochondrial particles (SMP) to establish the matrix or cytosolic orientation of loops M2 and M3. The results clearly show that these loops have a matrix orientation and thus support the six transmembrane segment model of ANT topography in the inner mitochondrial membrane. PMID- 12534296 TI - Oxidative dimer formation is the critical rate-limiting step for Parkinson's disease alpha-synuclein fibrillogenesis. AB - Intraneuronal deposition of alpha-synuclein as fibrils and oxidative stress are both implicated in the pathogenesis of Parkinson's disease. We found that the critical rate-limiting step in nucleation of alpha-synuclein fibrils under physiological conditions is the oxidative formation and accumulation of a dimeric, dityrosine cross-linked prenucleus. Dimer formation is accelerated for the pathogenic A30P and A53T mutant alpha-synucleins, because of their greater propensity to self-interact, which is reflected in the smaller values of the osmotic second virial coefficient compared to that of wild-type synuclein. Our finding that oxidation is an essential step in alpha-synuclein aggregation supports a mechanism of Parkinson's disease pathogenesis in which the separately studied pathogenic factors of oxidative stress and alpha-synuclein aggregation converge at the critical step of alpha-synuclein dimer formation. PMID- 12534297 TI - Formation of two peptide/MHC II isomers is catalyzed differentially by HLA-DM. AB - Major histocompatability class II proteins are transmembrane alphabeta heterodimers that present peptides to T-cells. MHC II may bind exogenous peptides directly at the cell surface. Alternatively, peptides derived from processing of endosomal protein may bind to MHC II in endosomal compartments. There, HLA-DM catalyzes the formation of peptide/MHC complexes, which are then transported to the cell surface. Here we report evidence that the peptide Ii CLIP 81-104 binds to DR*0404 in two alternate registries, whose dissociation rates, while kinetically indistinguishable at pH 5.3 and 37 degrees C, are kinetically resolved in the presence of HLA-DM. In one registry isomer, CLIP Met 91 is placed in the N-terminal P1 pocket of DR*0404, and peptide dissociation is readily catalyzed by HLA-DM. In a second proposed registry, likely with CLIP Leu 97 in the P1 pocket, the complex is substantially less sensitive to HLA-DM catalysis. Without HLA-DM, or at pH 7, the fraction of each isomer formed in solution is relatively insensitive to the duration of incubation with peptide. However, with HLA-DM, the fraction of the DM-insensitive isomer is dramatically influenced by peptide incubation time. The mechanism of isomer formation appears to be determined by the HLA-DM-modified relative association to the two registries, followed by HLA-DM-catalyzed dissociation of each isomer and rebinding, leading to a final isomer composition determined by these kinetic constants. Intramolecular isomer interconversion does not appear to be involved. The behavior of these complexes may provide a model for peptide editing by DM in endosomes. PMID- 12534299 TI - Cyclopentadienyl-carboranyl hybrid compounds: a new class of versatile ligands for organometallic chemistry. AB - A new class of organic-inorganic hybrid compounds bearing a cyclic pi-ligand, an icosahedral carborane moiety, and a bridging group has been developed. These are very versatile ligands, featuring both an organic and an inorganic moiety as well as a bridging ligand. They can be readily converted into mono-, di-, tri- and pentaanionic species under suitable reaction conditions, leading to the generation of a new class of organometallic compounds in which the carboranyl unit can be cast in the role of a bulky substituent, a sigma-, eta(5), eta(6), or eta(7) pi-ligand. This Account provides an overview of our recent work in this area. PMID- 12534301 TI - High-performance nanocatalysts for single-step hydrogenations. AB - Bimetallic nanoparticles (Ru(6)Pd(6), Ru(6)Sn, Ru(10)Pt(2), Ru(5)Pt, Ru(12)Cu(4), and Ru(12)Ag(4)) anchored within silica nanopores exhibit high activities and frequently high selectivities, depending upon the composition of the nanocatalyst, in a number of single-step (and often solvent-free) hydrogenations at low temperatures (333-373 K). The selective hydrogenations of polyenes (such as 1,5,9-cyclododecatriene and 2,5-norbornadiene) are especially efficient. Good performance is found with these nanoparticle catalysts in the hydrogenation of dimethyl terephthalate to 1,4 cyclohexanedimethanol and of benzoic acid to cyclohexanecarboxylic acid or to cyclohexene-1-carboxylic acid, and also in the conversion of benzene to cyclohexene (or cyclohexane), the latter being an increasingly important reaction in the context of the production of Nylon. Isolated atoms of noble metals (Pd, Rh, and Pt) in low oxidation states, appropriately complexed and tethered to the inner walls of nanoporous (ca. 3 nm diameter) silica, are very promising enantioselective hydrogenation catalysts. Nanoporous carbons, as well as other nanoporous oxides, may also be used to anchor and tether the kind of catalysts described here. PMID- 12534300 TI - Alpha-imino esters: versatile substrates for the catalytic, asymmetric synthesis of alpha- and beta-amino acids and beta-lactams. AB - The catalytic asymmetric addition of organic nucleophiles to alpha-imino esters has emerged as one of the most promising and intensely investigated routes to optically enriched alpha- and beta-amino acid derivatives and beta-lactams. The importance of alpha-imino esters stems not only from the vast appeal of the potential product classes,(1) but also from their remarkable reactivity as highly electrophilic imines. With each passing year, the number of publications concerning the asymmetric alkylation of imino esters grows significantly. The asymmetric alkylation of imines(2) and N,O-acetals has been in itself a subject of intense interest.(3) In this Account, we wish to illustrate our contribution to this timely field, as well as to highlight the seminal contributions of others. PMID- 12534302 TI - "Back to the future"--an account discovery of stable quasicrystals. AB - In this Account, I will describe the discoveries of a series of stable quasicrystals in which I participated during the past 15 years. These discoveries led to a paradigm shift in the field of quasicrystals. The story started when I was a graduate student, has continued into the present, and may continue into the future. Of these discoveries, some were made independently by myself and some in collaboration with my colleagues. As it happens, I am the only one related to all of these discoveries, and so it seems appropriate that I tell the stories. PMID- 12534303 TI - Supramolecular systems as microreactors: control of product selectivity in organic phototransformation. AB - This Account reviews the developments in microreactor-controlled selectivity in organic phototransformation. Photocycloaddition of alpha,omega-diaryl compounds with long flexible chains within Y-type zeolite and low-density polyethylene films leads to formation of intramolecular cyclomers to the exclusion of intermolecular products. ZSM-5 zeolite, Nafion membranes, and vesicles as hosts direct photosensitized oxidation of alkenes selectively toward either the energy transfer-mediated or the electron-transfer-mediated products. Zeolites, Nafion membranes, and microemulsions as microreactors remarkably control chemo-, regio-, and stereoselectivity in the photochemical reaction of phenyl phenylacetates, photocycloaddition of anthracenes, and photocyclization of azobenzene and stilbazole. PMID- 12534304 TI - Transition metal-catalyzed enantioselective ring-opening reactions of oxabicyclic alkenes. AB - Over the past five years, several metal-catalyzed asymmetric ring-cleaving reactions have been developed that generate ring-opened products in high yield and enantiomeric excess. These reactions can be carried out with a range of nucleophiles including hydride, stabilized and nonstabilized carbanions, alcohols, amines, and carboxylates. To achieve these results, three different transition metals have been employed, namely nickel, palladium, and rhodium, leading not only to synthetically useful transformations but also to a greater understanding of the reaction manifolds possible with these metals. This Account covers the work by our group and by others toward the development of new metal catalyzed asymmetric ring-opening reactions of oxabicyclic alkenes. PMID- 12534305 TI - Lepadiformine: a case study of the value of total synthesis in natural product structure elucidation. AB - Since the emergence of routine X-ray crystallography and high-field FT NMR in the mid-twentieth century, the importance of total synthesis in structure elucidation has become underappreciated by most organic chemists. However, the limitations and fallibility of spectral methodology has recently been highlighted by the mischaracterization of a number of complex natural products, the correct structures of which were all ultimately assigned by total synthesis. This account describes how total synthesis was not only instrumental in disproving the erroneously assigned structure of the marine alkaloid, lepadiformine, but also was also pivotal in establishing the correct structure and absolute configuration. PMID- 12534306 TI - Pi-nucleophilicity in carbon-carbon bond-forming reactions. AB - Which electrophiles react with which nucleophiles? The correlation log k(20 degrees Celsius) = s(E + N), in which electrophiles (carbocations, metal-pi complexes, diazonium ions) are characterized by one (E) and nucleophiles are characterized by two parameters (N, s), proved to be applicable for a wide variety of electrophile-nucleophile combinations. Since the introduction of this correlation in 1994 (Angew. Chem., Int. Ed. Engl. 1994, 33, 938-957), numerous new reagents have been characterized, and in 2001 (J. Am. Chem. Soc. 2001, 123, 9500-9512), a new method of parametrization was proposed that facilitates a continuous extension of the data sets without the need for reparametrization of existing data. This Account adjusts the N and s parameters of all presently characterized pi-nucleophiles (arenes, alkenes, organometallics) to the new parametrization and illustrates how to employ the resulting reactivity scales for analyzing synthetic and mechanistic problems in organic and macromolecular chemistry. Predictions of absolute rate constants, inter- and intramolecular selectivities, and analyses of reaction mechanisms are discussed. We outline how new compounds can be added to the scales and present our view on the scope and limitations of this approach to polar organic reactivity. PMID- 12534307 TI - [60]fullerene--metal cluster complexes: novel bonding modes and electronic communication. AB - [60]Fullerene can bind a variety of metal clusters via eta(2)-C(60), mu eta(2):eta(2)-C(60), and mu(3)-eta(2):eta(2):eta(2)-C(60) pi-type bonding modes. Multiple C(60) additions to a single cluster core have also been demonstrated. Modification of the coordination sphere of cluster moieties has resulted in novel transformation of the coordination mode of the C(60) ligand between pi and sigma (mu(3)-eta(1):eta(1):eta(2)-C(60) and mu(3)-eta(1):eta(2):eta(1)-C(60)) types as well as reversible interconversion between mu(3)-eta(2):eta(2):eta(2)-C(60) and mu-eta(2):eta(2)-C(60). The mu(3)-eta(2):eta(2):eta(2)-C(60) metal cluster complexes show remarkable electrochemical stability and an unusually strong electronic communication between C(60) and metal cluster centers. PMID- 12534310 TI - Effect of reproductive hormones and selective estrogen receptor modulators on mood during menopause. AB - Periods of intense hormonal fluctuations have been associated with heightened prevalence and exacerbation of underlying psychiatric illness, particularly the occurrence of premenstrual dysphoria, puerperal depression and depressive symptoms during perimenopause. It has been speculated that sex steroids such as estrogens, progestogens, testosterone and dehydroepiandrosterone (DHEA) exert a significant modulation of brain functioning, possibly through interactions with various neurotransmitter systems. It is therefore intuitive that abrupt alterations of these hormones would interfere with mood and behaviour. On the other hand, accumulating data suggest that hormonal interventions may also promote relief or even remission of depressive symptoms, as already demonstrated in studies with patients experiencing postpartum depression and perimenopausal depressive disorders. The extent to which perimenopause, alone, may increase the risk for depression is unclear. However, existing data strongly suggest that some women are particularly vulnerable to developing significant physical and psychological disturbances when entering perimenopause. This article reviews the effect of sex hormones and selective estrogen receptor modulators (SERMs) on mood among peri- and postmenopausal women. There are preliminary, though promising, data on the use of estradiol (particularly transdermal estradiol) to alleviate depression during perimenopause, use of a combination of estrogens and selective serotonin reuptake inhibitors for depression during the postmenopausal period, and the use of testosterone to improve psychological well-being and increase libido among women with induced menopause. Further studies would help to better delineate the usage of hormones as an antidepressant strategy (monotherapy or augmenting treatment) for peri- and postmenopausal women. A brief review of some nonhormonal interventions for the treatment of menopause-related symptoms that may significantly affect a woman's quality of life is also presented. There are some preliminary data suggesting the efficacy of antidepressants for the treatment of hot flushes; existing data on diet supplements and herbal products have shown more mixed results. PMID- 12534311 TI - Role of the immune system in the pathogenesis, prevention and treatment of Alzheimer's disease. AB - The dysregulation in the metabolism of beta-amyloid precursor protein and consequent deposition of amyloid-beta (Abeta) has been envisaged as crucial for the development of neurodegeneration in Alzheimer's disease (AD). Amyloid deposition begins 10-20 years before the appearance of clinical dementia. During this time, the brain is confronted with increasing amounts of toxic Abeta peptides and data from the last decade intriguingly suggest that both the innate and the adaptive immune systems may play an important role in the disorder. Innate immunity in the brain is mainly represented by microglial cells, which phagocytose and degrade Abeta. As the catabolism of Abeta decreases, glial cells become overstimulated and start to produce substances that are toxic to neurons, such as nitric oxide and inflammatory proteins. Pro-inflammatory cytokines can be directly toxic or stimulate Abeta production and increase its cytotoxicity. A therapeutic possibility arises from clinical studies, which demonstrate that nonsteroidal anti-inflammatory drugs (NSAIDs) may delay the onset and slow the progression of AD. Recent data show that in addition to the suppression of inflammatory processes in the brain NSAIDs may decrease the production of Abeta peptides. The role of adaptive immunity lies mainly in the fact that Abeta can be recognised as an antigen. Immunisation with Abeta peptides and peripheral administration of Abeta-specific antibodies both decrease senile plaques and cognitive dysfunction in murine models of AD. A recent trial in humans seems still to be hampered by adverse effects. As adaptive immunity decreases with aging while innate immunity remains intact, immunotherapy for AD will have to be adapted to this situation. Strategies that combine vaccination and inflammatory drug treatment could be considered. PMID- 12534312 TI - Value of endocrine therapy for early and locally advanced prostate cancer. AB - Prostate-specific antigen-based screening and increased public awareness of prostate cancer have led to earlier detection of localised and potentially curable tumour stages. Nevertheless, disease recurrence after radical treatment of prostate cancer remains a problem for a considerable number of patients. Endocrine therapy plays a role in the treatment of almost all stages of prostate cancer, either as definitive or as temporary therapy. Neoadjuvant hormonal treatment has its place mainly in the external beam radiotherapy setting, where it reduces target volume and adverse effects. When used prior to and during external beam radiotherapy, endocrine therapy has shown a survival advantage in the subset of patients with locally advanced prostate cancer (cT2-4, N0-1, M0) with Gleason scores 2-6. Several studies have shown that adjuvant endocrine treatment is able to delay disease progression in any stage. There is, however, an ongoing controversy regarding the possible survival benefit of such treatment that has to be balanced against its considerable adverse effects and costs. Survival improvements associated with adjuvant hormonal treatments have mainly been seen in patients with lymph node metastases or those with a high probability of micrometastatic disease. Several large studies are under way to investigate the role of adjuvant treatment in the field of early prostate cancer. PMID- 12534313 TI - Selecting neuromuscular-blocking drugs for elderly patients. AB - The physiological changes that occur with increasing age can have significant effects on the pharmacokinetics of neuromuscular-blocking drugs. Changes in cardiac output can affect drug distribution and therefore the speed of onset of neuromuscular block. A decrease in muscle mass and increase in body fat with age can also affect their distribution. The deterioration in renal and hepatic function associated with aging affects the clearance and elimination of many neuromuscular-blocking drugs. The effects of these physiological changes on the pharmacokinetics of neuromuscular-blocking agents may not become apparent clinically in healthy individuals until the age of at least 75 years. There is very little evidence to suggest any alteration in the sensitivity of the neuromuscular junction to neuromuscular-blocking drugs with increasing age. Neuromuscular-blocking drugs that undergo a significant degree of organ-dependent elimination, such as pancuronium bromide, vecuronium bromide, rocuronium bromide and doxacurium chloride, may have a significantly prolonged duration of action in elderly patients. These drugs can be used safely in elderly patients if the anaesthetist is aware of their altered pharmacokinetics in this patient group. Appropriate changes must be made to drug dosage and dose intervals. As the pharmacokinetic changes can be unpredictable, monitoring of neuromuscular block is strongly advised when using these drugs in such patients. The risk of residual block occurring postoperatively after the use of pancuronium bromide increases with age. The duration of action of mivacurium chloride may also be prolonged in the elderly; this change has not been demonstrated to be a result of an alteration in plasma cholinesterase activity. In contrast, there is no evidence of an alteration in the action of suxamethonium chloride (succinylcholine chloride) with increasing age. Atracurium besilate and cisatracurium besilate undergo predominantly organ-independent elimination. Onset of block with these two drugs may be prolonged in the elderly, but their clinical duration of action does not alter significantly with age, making them particularly suitable for use in this patient group. Although atracurium besilate may cause histamine release, there is little evidence of it producing haemodynamic changes in the elderly. Its (1R,1R')-isomer, cisatracurium besilate, has very little direct or indirect cardiovascular effect and is, therefore, the most suitable nondepolarising agent to use in elderly patients. PMID- 12534314 TI - Drug treatment of epilepsy in elderly people: focus on valproic Acid. AB - Despite old age being the commonest time of life to develop epilepsy, relatively little is known about the condition in later years. Antiepileptic drugs (AEDs) are the mainstay of treatment and valproic acid (VPA) has been prescribed for older patients with seizures for over 35 years. VPA is available in a variety of formulations. The drug is generally rapidly absorbed, although there are no data on the extent of oral absorption in the elderly. The volume of distribution (Vd) and elimination half-life have been compared in older and younger patients. One study reported no change in either parameter between elderly and younger patients (Vd: 0.16 vs 0.14 L/kg; elimination half-life: 15.3 vs 13.0h), the other found an increase in both for older patients (Vd: 0.19 vs 0.13 L/kg; elimination half-life 14.9 vs 7.2h). Total VPA clearance is similar in young and elderly subjects. The drug does not induce the metabolism of hepatic enzymes, but can act as a metabolic inhibitor, raising plasma concentrations of lamotrigine, phenobarbital (phenobarbitone), carbamazepine-10-11-epoxide, lorazepam, nimodipine and zidovudine. Concomitant use of VPA may also lead to an elevation in phenytoin, diazepam, warfarin, amitriptyline and chlorpromazine concentrations. A number of enzyme-inducing AEDs such as phenytoin, phenobarbital, primidone and carbamazepine can increase the clearance of VPA. Plasma concentrations of VPA may also rise when the drug is administered with felbamate, stiripentol, aspirin (acetylsalicylic acid), naproxen, phenylbutazone, isoniazid, fluoxetine and chlorpromazine. The majority of elderly patients present with partial and/or secondary generalised seizures, although a few have long-standing primary generalised seizures. Results from meta-analyses and randomised studies of patients comparing VPA with other AED monotherapies suggest that the drug is as effective as carbamazepine, phenytoin and phenobarbital in treating these seizure types. Although some of these studies recruited older patients, there have been no randomised double-blind trials examining the efficacy of VPA with other AEDs in an exclusively elderly cohort. There is no direct correlation between efficacy and plasma VPA concentrations. The majority of older patients require lower doses of AEDs than younger adults. Higher VPA doses may be needed in patients taking drugs which induce hepatic microsomal enzymes. Once-daily dosing of the controlled-release preparation can help to improve compliance and may render some frail elderly people seizure free. There is a perception that the elderly are generally more susceptible to the adverse effects of AEDs than younger adults, although there are few data to validate this claim. Dose-dependent and idiosyncratic reactions may be more frequent. Common adverse effects of VPA include gastrointestinal symptoms and tremor. Slow-dose escalation and controlled release preparations may minimise these. In summary, VPA is a long established AED. Its broad spectrum of action and dosing schedule are favourable properties for its use in older people. To accurately establish the place of this and other AEDs in treating elderly patients with epilepsy, well designed clinical trials are urgently required in this vulnerable population. PMID- 12534315 TI - Comparison of the efficacy and tolerability of policosanol with atorvastatin in elderly patients with type II hypercholesterolaemia. AB - BACKGROUND: Hypercholesterolaemia is a risk factor for coronary heart disease (CHD). Clinical studies have shown that lowering elevated serum total cholesterol (TC) levels, and particularly low density lipoprotein-cholesterol (LDL-C) levels, reduces the frequency of coronary morbidity and deaths, whereas high serum levels of high density lipoprotein-cholesterol (HDL-C) protect against CHD. Policosanol is a cholesterol-lowering drug purified from sugar cane wax with a therapeutic dosage range from 5-20 mg/day. Atorvastatin is an HMG-CoA reductase inhibitor which across its dosage range (10-80 mg/day) has shown significantly greater lipid-lowering effects than all previously marketed statins. OBJECTIVE: This study was undertaken to compare the efficacy and tolerability of policosanol with atorvastatin in older patients with type II hypercholesterolaemia. PATIENTS AND METHODS: This randomised, single-blind, parallel-group study was conducted in older patients (60-80 years) with type II hypercholesterolaemia. After 4 weeks on a cholesterol-lowering diet, 75 patients were randomised to policosanol or atorvastatin 10mg tablets taken once daily with the evening meal for 8 weeks. An interim and final check-up were performed at 4 and 8 weeks, respectively, after treatment was initiated. RESULTS: At 4 (p < 0.0001) and 8 (p < 0.00001) weeks, policosanol 10 mg/day significantly lowered serum LDL-C levels by 17.5 and 23.1%, respectively compared with baseline; corresponding values for atorvastatin were 28.4 and 29.8%. At study completion, policosanol significantly (p < 0.0001) reduced serum TC (16.4%), LDL-C/HDL-C ratio (25.5%) and TC/HDL-C ratio (19.3%), as well as (p < 0.001) triglyceride levels (15.4%). Atorvastatin significantly (p < 0.0001) decreased serum TC (22.6%), LDL-C/HDL-C (26.2%) and TC/HDL-C (19.8%) ratios, as well as (p < 0.001) triglyceride levels (15.5%). Atorvastatin was significantly more effective than policosanol in reducing LDL-C and TC, but similar in reducing both atherogenic ratios and triglyceride levels. Policosanol, but not atorvastatin, significantly (p < 0.05) increased serum HDL-C levels by 5.3%. Both treatments were well tolerated. At study completion, atorvastatin mildly, but significantly (p < 0.05) increased creatine phosphokinase (CPK) and creatinine, whereas policosanol significantly reduced AST and glucose (p < 0.01) and CPK (p < 0.05) levels. All individual values, however, remained within normal limits. Three atorvastatin but no policosanol patients withdrew from the study because of adverse events: muscle cramps (1 patient), gastritis (1 patient) and uncontrolled hypertension, abdominal pain and myalgia (1 patient). Overall, no policosanol and seven atorvastatin patients (18.9%) reported a total of nine mild or moderate adverse events during the study (p < 0.01). CONCLUSIONS: This study shows that policosanol (10 mg/day) administered for 8 weeks was less effective than atorvastatin (10 mg/day) in reducing serum LDL-C and TC levels in older patients with type II hypercholesterolaemia. Policosanol, but not atorvastatin, however, significantly increased serum HDL-C levels, whereas both drugs similarly reduced atherogenic ratios and serum triglycerides. Policosanol was better tolerated than atorvastatin as revealed by patient withdrawal analysis and overall frequency of adverse events. Nevertheless, further studies must be conducted in larger sample sizes and using dose-titration methods to achieve target lipid levels in order to reach wider conclusions. PMID- 12534316 TI - Interleukin-12: potential role in asthma therapy. AB - Asthma is an inflammatory disease of the airways leading to significant morbidity and mortality. With advances in the understanding of the molecular and cellular mechanisms involved in the asthmatic response, researchers have identified specific mediators that may be targeted to control the inflammatory state of asthma. The Th2 hypothesis proposes that the inflammation in asthma arises from an imbalance between the two CD4+ T lymphocyte subsets, T helper (Th) type 1 and Th2. Th2 cells release many cytokines that have been shown to regulate the inflammatory response, while the Th1 cytokines counteract this response. The Th1 cytokine, interleukin (IL)-12, has been a target of intense study because it mediates the Th1 response and offers a means of modifying the asthmatic inflammatory response. Numerous murine studies have shown that this cytokine can potently inhibit allergic airway inflammation in asthma. Inhalation of IL-12 has been shown to increase its efficacy in inhibiting allergic inflammation in murine models while decreasing adverse effects seen with systemic administration of this cytokine. However, an initial study of inhaled IL-12 in humans with asthma was terminated because of adverse effects. The use of systemically administered IL-12 in patients with asthma has been limited due to cytokine toxicity. Another treatment option that has the potential of inducing a Th1 cytokine response is the use of IL-12 linked to polyethylene glycol (PEG) moieties. This mode of administration is likely to enhance cytokine delivery to the target organ, while decreasing its toxicity. IL-12 gene therapy has also been examined as a means of suppressing airway hyperreactivity in murine asthma, but its potential in human asthma has not been explored. Several recent studies have investigated the role of CpG DNA motifs as endogenous inducers of IL-12 with encouraging results in both mice and humans. These studies may result in novel Th1- inducing CpG-based immunotherapies for asthma. PMID- 12534317 TI - Recent advances in the development of an inhaled insulin product. AB - Inhaled insulin first entered clinical human testing in the mid 1990s. Since then, the commercial potential and technical challenges of an inhaled insulin product have grown increasingly clear, with several pharmaceutical partnerships now targeting treatment of diabetes mellitus through inhalation products in clinical development. While clinical results to date show the therapy to be generally promising, recent data have raised questions related to human safety and slowed progress toward a commercial product. Answering these questions positively in the coming years will be critical to making inhalation therapy a practical diabetes-care reality. PMID- 12534318 TI - Recombinant human thyroid-stimulating hormone: pharmacology, clinical applications and potential uses. AB - The major functions of pituitary thyroid-stimulating hormone (TSH) are to maintain the biosynthesis and secretion of the thyroid hormones L-thyroxine (T4) and L-3,5,3'triidothyronine (T3). The TSH core contains two apoproteins, the alpha and beta subunits. The alpha subunit is identical to that of pituitary follitropin, pituitary lutropin and placental chorionic gonadotropin, whereas the beta subunit is unique. TSH is a glycoprotein; the glycoprotein components of the alpha and beta subunits account for more than 10% of their mass and are essential for normal thyrotropic action and intravascular kinetics. The hypothalamic tripeptide, TSH-releasing hormone (TRH) is required for optimum TSH biosynthesis, particularly as far as addition of the glycoprotein components is concerned. TRH deficiency is associated with secretion of TSH molecules that are appropriately measured in most assays but have reduced bioactivity. In previous years the TSH used in clinical practice was obtained and purified from bovine pituitaries. Bovine TSH was used to test thyroid function and to augment the uptake of radioiodine in patients with thyroid cancer. Bovine TSH has been largely abandoned as a clinical agent because of adverse immune reactions. A recombinant human TSH (rhTSH; Thyrogen), has been approved by the US FDA for diagnostic use in patients with thyroid cancer. The alpha and beta subunits of Thyrogen are identical to those of human pituitary TSH. Thyrogen has a specific activity of approximately 4 IU/mg and is a potent stimulator of T4, T3 and thyroglobulin (Tg) secretion in healthy volunteers. It also increases thyroid iodide uptake in patients with thyroid cancer or multinodular goitre and in volunteers, even those exposed to large amounts of stable iodide. Thyroid cancer patients who have been treated by thyroidectomy and radioiodine ablation but are at risk of harbouring residual thyroid cancer are candidates for Thyrogen administration to prepare them for whole body iodide scans and serum Tg measurements. In thyroidectomised thyroid cancer patients who are unable to secrete pituitary TSH upon thyroid hormone withdrawal, Thyrogen is the only acceptable method to prepare them for these procedures. Thyrogen has been used on a compassionate basis to prepare patients for radioiodine ablation. rhTSH, in addition to being useful in the management of patients with thyroid cancer, is potentially useful to test thyroid reserve and to aid in thyroid-related nuclear medicine procedures. In the future, TSH analogues that have superagonist or antagonist properties may become available as therapeutic agents. PMID- 12534319 TI - Prevention strategies for type 1 diabetes mellitus: current status and future directions. AB - Type 1 diabetes mellitus affects about 1 in 300 people in North America and Europe. Epidemiological studies indicate that the incidence and thus prevalence of type 1 diabetes is rising worldwide. Intervention in autoimmune type 1a diabetes could occur at the time of diagnosis or, preferably, prior to clinical presentation during the 'prediabetic' period (e.g. prevention). Prediabetes is best recognised by the detection of islet autoantibodies in the serum. Promising intervention strategies include monoclonal antibody therapies (e.g. anti-CD3, anti-CD25, anti-CD52 or anti-CD20 monoclonal antibodies), immunosuppression (e.g. calcineurin inhibitors, B7 blockade, glucocorticoids, sirolimus (rapamycin), azathioprine or mycophenolate mofetil), immunomodulatory therapies (e.g. plasmapheresis, intravenous immunoglobulin, cytokine administration, adoptive cellular gene therapy) and tolerisation interventions (e.g. autoantigen administration or avoidance, altered peptide ligand or peptide-based therapies). To date, islet and pancreas transplantation have essentially been reserved for patients with long-standing diabetes who have complications and are also in need of a concurrent kidney transplant. None of the therapies attempted to date has produced long-term remissions in new-onset type 1 diabetes patients and no therapies have been shown to prevent the disease. Nevertheless, with advances in our understanding of basic immunology and the cellular and molecular mechanisms of tolerance induction and maintenance, successful intervention therapies will be developed. The balance between safety and efficacy is critical. Higher rates of adverse events might be more tolerable in new-onset type 1 diabetes patients if the therapy is extremely effective at inducing a permanent remission. However, therapies must not harm the beta-cells themselves or any organ system that is a potential target of diabetes complications, such as the nervous system, retina, cardiovascular system or kidney. In the treatment of prediabetes, successful therapies should provide a level of safety similar to that of currently used vaccines and a high level of efficacy. PMID- 12534321 TI - Amotosalen: Allogeneic Cellular Immunotherapies system, INTERCEPT Plasma System, INTERCEPT Platelet System, S 59. AB - Adis CommentsCerus Corporation is developing a variety of pathogen-inactivation systems, based on its Helinx technology. Three of the systems include amotosalen [S 59] as the inactivation compound. Amotosalen is a light-activated, DNA-, RNA crosslinking psoralen compound, which is used to neutralise pathogens. The systems that utilise amotosalen are called the INTERCEPT Platelet System, the INTERCEPT Plasma System and the Allogeneic Cellular Immunotherapies (ACIT) system. The INTERCEPT Platelet System and INTERCEPT Plasma System are two of the systems that make up Cerus' INTERCEPT Blood Systems. The other system is the INTERCEPT Red Blood Cell System, which contains S 303 as the inactivation compound rather than amotosalen. Cerus' Helinx technology is able to prevent replication of DNA or RNA that is present in pathogens but not in the blood components being treated (e.g. platelets and plasma). When added to the blood components, the inactivation agent (in this case amotosalen) crosses the membrane or cell wall of the pathogen. When activated by light, amotosalen binds to the nucleic acid of the pathogen and prevents replication. This process prevents infection. INTERCEPT Platelet System: Cerus developed its INTERCEPT Platelet System, in collaboration with Baxter Healthcare, for use in blood centres. Platelets are an essential component of the coagulation process and may be required by patients undergoing surgery, cancer chemotherapy, transplantation or with bleeding disorders. The system is made up of an illuminator device, a compound absorption device and a processing kit containing amotosalen. In October 2002, the two companies announced that CE Mark approval had been received for the illuminator device for the INTERCEPT trade mark Blood System. Application of this technology to platelets is the first to be approved. As it is a new technology, the system is currently undergoing process validation in accordance with European Blood Bank GMP requirements. This validation process is currently being conducted in Denmark, France, Germany, Sweden and the UK. Marketing approval applications for the INTERCEPT Platelet System have also been submitted in Australia and Canada. In addition, the regulatory submission process has begun in the US. A phase III trial (EuroSPRITE) has been conducted in 103 patients in Europe with pooled random donor platelets. The platelets were collected using the buffy coat process. Another two 20-patient clinical trials have also been conducted in Europe, as well as a 40-patient trial using platelets collected by an apheresis collection system. Cerus has also conducted a phase III trial (SPRINT) in the US. The trial was conducted in 671 patients and used platelets collected by Baxter's apheresis collection system. INTERCEPT Plasma System: Cerus is also developing the INTERCEPT Plasma System in collaboration with Baxter Healthcare. The system also combines amotosalen, an illumination device and a compound absorption device. The two companies are currently preparing regulatory applications for the INTERCEPT Plasma System for the US. This application will be followed by a submission for CE Mark designation in Europe. Patients undergoing surgery, or transplantation, or with bleeding disorders, may require transfusions of plasma, often to control bleeding. The type of plasma is stored in frozen form and is called fresh frozen plasma (FFP). The INTERCEPT Plasma System is currently in phase IIIc development in the US. Patient enrolment in the trial is still ongoing. The trial is comparing INTERCEPT trade mark Plasma System treated versus untreated FFP in 30 patients with thrombotic thrombocytopenic purpura. Allogeneic Cellular Immunotherapies system: Cerus is also investigating the potential of its Helinx technology to improve the outcome of bone marrow transplantation procedures (used to treat leukaemia and lymphoma) through the treatmatment for many forms of leukaemia and is most effective when the donor is very closely matched to the patient for the major human leucocyte antigen (HLA) groups. As part of the transplant procedure, patients receive donor T cells to improve engraftment of the bone marrow transplant and strengthen the patient's immune system. However, donor T cells expose the patient to a high risk of contracting graft-versus-host disease (GVHD) caused by the proliferation of donor T cells, which attack the patient's healthy tissue. GVHD has a high mortality rate. Cerus' ACIT system has been developed to decrease the stringency of matching donors to patients and to inhibit the ability of donor T cells to cause GVHD. Light activated amotosalen binds and permanently crosslinks DNA, preventing replication and thus stopping proliferation of donor T cells. Phase I development is currently being conducted in this area in the US using amotosalen as the neutralising agent. Cerus completed a phase I study investigating the safety and tolerability of its ACIT system in 2001. The study was conducted in patients receiving closely matched allogeneic bone marrow transplants for leukaemia. The company is currently collaborating with the National Marrow Donor Program in order to conduct further clinical studies in patients receiving bone marrow transplants from unmatched donors. Cerus has development, manufacturing and marketing agreements with Baxter covering the INTERCEPT Blood Systems, which includes the INTERCEPT Platelet system, the INTERCEPT Plasma System, and the INTERCEPT Red Blood Cell System. Under the terms of the agreements the two companies usually share the very early development activities. Cerus then conducts preclinical and clinical trials, while Baxter is responsible for the development of the systems disposables and devices. Following commercialisation Cerus will supply amotosalen and Baxter will supply the other components of the system and market, sell and distribute the system In January 2001, Cereus announced that it has entered into a collaborative agreement with the Pharmaceutical Division of Kirin Brewery in Japan to develop and market products for stem cell transplantation based on Cerus' proprietary Helinx technology. Under terms of the agreement, Cerus and Kirin will jointly develop the products. Cerus has received an initial license fee of US dollar 1 million. In addition it may receive up to US dollar 11 million in future payments upon achievement of development milestones. Kirin will also fund all development expenses for the Asia-Pacific region and a portion of Cerus' development activities aimed at obtaining product approval in the US. Kirin will market the products in the Asia Pacific region, including Japan, China, Korea and Australia, and Cerus will receive a specified share of product revenues. Cerus will retain marketing rights in the rest of the world, including the US and Europe. PMID- 12534322 TI - Melanoma Vaccine--AVAX Technologies: DNP-VACC, M-Vax. AB - Adis CommentsAVAX Technologies is developing a therapeutic melanoma vaccine [M Vax, DNP-VACC] consisting of autologous tumour cells conjugated to a highly immunogenic hapten, dinitrophenyl, which makes the cancer cells more easily recognised by the immune system. AVAX licensed the autologous cell vaccine technology (AC Vaccine) from Thomas Jefferson University in Philadelphia, USA, where it was originally developed. M-Vax was launched in Australia in the first half of 2000, but was withdrawn from this market in September 2002 due to financial constraints faced by the company and its need to focus its resources on initiatives that provide the greatest return. Although AVAX applied for Federal Government price reimbursement in Australia through the Medical Services Advisory Committee during 2001, the vaccine is not reimbursed in Australia. Obtaining Federal Government reimbursement was a step AVAX considered essential for the success of the M-Vax. AVAX has not ruled out re-entering the Australian market again at a later date. AVAX will now concentrate on gaining approval in the US and Europe. M-Vax has received orphan drug designation from the US FDA. M-Vax is in preregistration in Germany, Japan and The Netherlands for treatment of stage III melanoma. In September 1999, the company announced that it expected to market M-Vax for treatment of stage III melanoma in Germany, Japan and the Netherlands. This announcement came after AVAX's continuing dialogue with senior regulatory authorities in several pharmaceutical markets. The commercial availability of M Vax in Germany, Japan and The Netherlands will be subject to meeting certain requirements specified by the regulatory agency in each country. Phase II data have been submitted for regulatory approval in these countries; phase III data may not be required because the vaccine contains autologous tumour cells. This was the case with the Australian approval of M-Vax, which was on the basis of data from phase II trials. Clinical development: M-Vax was in a pivotal phase III trial for treatment of stage III melanoma in the US, and a multicentre phase II trial in the US for treatment of patients with stage IV melanoma with lung metastases. However, in late March 2001, AVAX announced that the FDA had suspended these trials until the agency had further reviewed them. Subsequently, AVAX received written communication from the FDA indicating that the suspension is related to manufacturing issues. These events triggered the resignations of AVAX's executive Vice-President and Vice-President of operations, at the request of the company's board of directors. AVAX met with the FDA in October 2001 to discuss the clinical holds on M-Vax and O-Vax. AVAX's proposed improvements involving a frozen vaccine were also discussed at the meeting. Following the meeting AVAX was told by the FDA that selected characterisation work would have to be carried out on the new products, and new INDs submitted. In December 2001 AVAX announced that the development of a frozen vaccine and changes to various policies and procedures would ensure that the company complied with the FDA regulations. A new IND was submitted to the FDA for M-Vax in September 2002. In August 2002, AVAX had been unsure whether following approval of its new IND it would re-initiate clinical development for both M-Vax and O-Vax in parallel, or advance one of the agents and wait for further funding for the other. However, in September it indicated that clinical trials of both vaccines would be conducted following approval of the IND. A total of 42 patients are to be enrolled in each trial. In October 2002, AVAX announced that the US FDA had no outstanding issues regarding the IND. AVAX can now proceed with clinical trials as planned. AVAX Technologies was enrolling patients with stage III melanoma in the pivotal US phase III trial for registration of M-Vax trade ed, multicentre trial designed to compare the efficacy of the vaccine against high-dose interferon-alpha, the standard post-surgical treatment for stage III melanoma. The two end-points are rate of melanoma tumour recurrence and overall survival. The dosing regimen chosen for this study is that which was found from several clinical studies to be most effective at eliciting a positive delayed-type hypersensitivity skin response to autologous melanoma cells. The study was being conducted at more than 20 US sites. A low dose of M-Vax was also being evaluated in a phase II study at Thomas Jefferson University in the US. On 16 March 2000, AVAX announced promising interim results from this study, which revealed that 65% of 23 evaluated patients developed an immune response of the same magnitude as that observed with higher doses of M-Vax in previous studies. The study was to enroll a total of 46 patients, who were to receive seven doses of M-Vax over 7 weeks. The advantage of using a low dose of M-Vax is that it requires a smaller amount of the patient's tumour tissue to produce the vaccine (approximately half that required in previous studies) and therefore more patients would be eligible for treatment. On the basis of these results, AVAX modified the pivotal phase III trial to use the low dose of M-Vax to treat additional patients with smaller tumours. On 29 March 2000, AVAX announced that it had initiated a multicentre phase II study in the US in patients with stage IV melanoma and lung metastases. Patients were receiving seven doses of M-Vax at weekly intervals and a booster at 6 months. AVAX initiated the study because of promising results in a study of stage IV melanoma patients with lung metastases in which patients treated with M-Vax had tumour regression and prolonged survival. Commercial agreements: In June 1999, AVAX announced its first international commercialisation opportunity for M-Vax, in Australia, where the company subsequently launched the vaccine (now withdrawn) in the first half of 2000. AVAX formed a subsidiary, AVAX Australia, which was co marketing M-Vax in Australia together with Australian Vaccine Technologies (formerly Neptunus International Holdings). Under the terms of this agreement, Australian Vaccine Technologies purchased dollars A10 million in shares, a 50% interest, in AVAX Australia. The final dollars A3 million installment was made in August 2000. AVAX had an option to purchase up to 5% of shares in Australian Vaccine Technologies. In August 2002, AVAX extended and expanded an existing production agreement with Medigene for approximately 1 year. Under the terms of the agreement, Genopoietic (Medigene) in France will process clinical samples of M-Vax. In October 2002, AVAX signed a distribution agreement with Ferrer International, SA (Grupo Ferrer) for sales and distribution of the AC Vaccines, including M-Vax and O-Vax. The agreement covers Europe, Latin America and certain Asian territories. Under the terms of the agreement AVAX will retain manufacturing rights and will sell the vaccine to Ferrer. In return, Ferrer will make payments to AVAX for the product as well as certain milestone payments for marketing and registration goals. M-Vax was manufactured in Australia by Bioenterprises, a subsidiary of Biotech Australia. However, in 2002, the manufacturer underwent an acquisition with significant changes, which resulted in its decision to discontinue manufacturing M-Vax. PMID- 12534323 TI - Pramlintide: (AC 137, AC 0137, Symlin, Tripro-Amylin). AB - Adis CommentsPramlintide [AC 0137, AC 137, tripro-amylin, Symlin] is a synthetic human amylin analogue with proline substitutions at positions 25, 28 and 29, which limits the self-aggregation seen with native amylin. Pramlintide improves glycaemic control, and appears to reduce postprandial blood glucose peaks and flatten the glucose peaks and troughs observed in diabetic patients. The reduction of hypoglycaemia would be an immediate advantage, and the reduction of hyperglycaemia could potentially prevent diabetic complications. Development - US: Amylin has submitted an NDA in the US for pramlintide acetate (Symlin trade mark ) as an adjunctive therapy for the treatment of type 1 and type 2 diabetes mellitus. However, the FDA's Endocrinologic and Metabolic Drugs Advisory Committee at their meeting on 26 July 2001, voted not to recommend approval of pramlintide for type 1 and type 2 diabetes. Although eight out of nine Committee members were convinced of the potential of pramlintide therapy, the Committee expressed concerns regarding safety issues and requested additional data addressing these concerns. Finally, on 12 October 2001, Amylin received an 'approvable letter' for Symlin- for the treatment of diabetes. In April 2002, Amylin commenced a trial in 250 patients with type 1 diabetes to evaluate the safety issues regarding cases of severe hypoglycaemia with pramlintide in combination with insulin reported in this group of patients. The trial will investigate dose titration in the initial first month of the treatment period combined with insulin adjustment for the optimisation of glucose control. Patients are then treated for 6 months at a steady-state dose of pramlintide or placebo, accompanied by the additional insulin adjustments. Amylin has completed patient enrolment in September 2002. Final approval is subject to satisfactory results from this safety and dose titration study and the four small pharmacology studies already completed or underway. Amylin plans to file an amendment to the pramlintide's NDA in the Q1 of 2003. Development - non-USA: A wholly owned subsidiary of Amylin Pharmaceuticals, Amylin Europe, filed a regulatory submission with the European Agency for Evaluation of Medicinal Products (EMEA) and Switzerland for pramlintide for the treatment of both type 1 and type 2 diabetes under the centralised procedure. Amylin completed pivotal phase III clinical trials with pramlintide acetate (Symlin trade mark ) for the treatment of type 1 and type 2 diabetes mellitus in North America and Europe. However, in October 2002, Amylin announced that following consultation with the Committee for Proprietary Medicinal Products (CPMP) of the EMEA, it has found that additional information is necessary to proceed with review of the MAA for pramlintide for diabetes. Since, the centralised procedure does not allow the adding of new information to the application that is already under review, Amylin has decided to withdraw the MAA for pramlintide. The company will continue discussions with the EMEA to clarify the information required for a resubmission of the application. The submission for pramlintide in Switzerland is currently under review. In a separate phase II programme, Amylin is investigating the use of pramlintide in type 2 diabetes mellitus patients who are not achieving satisfactory results with oral hypoglycaemic agents but who have not progressed to using insulin. Collaborations: Pramlintide was under joint development with Amylin Pharmaceuticals and Johnson and Johnson, as an injectable partner hormone for insulin for the treatment of both type 1 and type 2 diabetes mellitus. The terms of the agreement between Amylin and Johnson and Johnson were that Amylin had primary responsibility for development and regulatory submissions, while Johnson and Johnson had primary responsibility for marketing; development costs and eventual profits were to be shared equally. Later, Johnson and Johnson decided to terminate the collaboration to commercialise pramlintide. An earlier development collaboration betweion between Amylin and Glaxo Wellcome was also discontinued. However, Amylin is in new ongoing discussions with collaborative partners for pramlintide in Europe and Japan. Amylin has signed an agreement with CP Pharmaceuticals in the UK to manufacture pramlintide. PMID- 12534324 TI - Management of calcium channel antagonist overdose. AB - Calcium channel antagonists are used primarily for the treatment of hypertension and tachyarrhythmias. Overdose of calcium channel antagonists can be lethal. Calcium channel antagonists act at the L-type calcium channels primarily in cardiac and vascular smooth muscle preventing calcium influx into cells with resultant decreases in vascular tone and cardiac inotropy and chronotropy. The L type calcium channel is a complex structure and is thus affected by a large number of structurally diverse antagonists. In the setting of overdose, patients may experience vasodilatation and bradycardia leading to a shock state. Patients may also be hyperglycaemic and acidotic due to the blockade of L-type calcium channels in the pancreatic islet cells that affect insulin secretion. Aggressive therapy is warranted in the setting of toxicity. Gut decontamination with charcoal, or whole bowel irrigation or multiple-dose charcoal in the setting of extended-release products is indicated. Specific antidotes include calcium salts, glucagon and insulin. Calcium salts may be given in bolus doses or may be employed as a continuous infusion. Care should be exercised to avoid the administration of calcium in the setting of concomitant digoxin toxicity. Insulin administration has been used effectively to increase cardiac inotropy and survival. The likely mechanism involves a shift to carbohydrate metabolism in the setting of decreased availability of carbohydrates due to decreased insulin secretion secondary to blockade of calcium channels in pancreatic islet cells. Glucose should be administered as well to maintain euglycaemia. Supportive care including the use of phosphodiesterase inhibitors, adrenergic agents, cardiac pacing, balloon pump or extracorporeal bypass is frequently indicated if antidotal therapy is not effective. Careful evaluation of asymptomatic patients, including and electrocardiogram and a period of observation, is indicated. Patients ingesting a nonsustained-release product should be observed in a monitored setting for 12 hours, while those who ingest a sustained-release preparation should be observed for no less than 24 hours. Charcoal should be given to the asymptomatic patient with a history of calcium channel antagonist overdose. PMID- 12534325 TI - Fenofibrate-induced hyperhomocysteinaemia: clinical implications and management. AB - Fenofibrate is among the drugs of choice for treatment of hypertriglyceridaemia and low levels of high-density lipoprotein (HDL)-cholesterol, both recognised as risk factors for cardiovascular disease. Recently, a number of studies have shown an elevation of homocysteine levels with fenofibrate or bezafibrate therapy. Homocysteine is an atherogenic amino acid derived from the methionine cycle. At present, the underlying mechanism for this elevation has not been elucidated. While deterioration of vitamin status does not seem to be involved, impairment of renal function or changes in creatine metabolism are regarded as probable mechanisms. In patients not receiving lipid-lowering drugs, vitamin supplementation with folic acid and vitamin B12 effectively reduces the plasma homocysteine level. Two studies have shown that addition of folic acid or a vitamin combination to fenofibrate prevented most of the homocysteine increase associated with fenofibrate. Although the consequence of increasing homocysteine levels for cardiovascular risk has not been proven at present, it has to be considered that fenofibrate will be given for long-term treatment. Therefore, addition of folic acid and vitamin B12 to fenofibrate can be recommended to prevent the increase of homocysteine associated with fenofibrate, or treatment could be changed to gemfibrozil, which does not increase plasma homocysteine levels. PMID- 12534327 TI - Adverse drug reactions related to the use of fluoroquinolone antimicrobials: an analysis of spontaneous reports and fluoroquinolone consumption data from three italian regions. AB - OBJECTIVE: To analyse and compare with one another and with other antibacterial drugs the adverse drug reactions (ADRs) of the different fluoroquinolones currently used in Italy, spontaneously reported from doctors in three northern Italian regions. METHODS: The data on fluoroquinolones and other antibacterials were obtained from the spontaneous reporting system database of Emilia Romagna, Lombardy and the Veneto, which are the principal contributors to the Italian spontaneous surveillance system. The fluoroquinolone ADRs with a causality assessment of certain, probable or possible (according to WHO criteria), reported between January 1999 and December 2001, were selected and toxicity profiles of individual drugs were described and compared with one another. The reports were also correlated with sex and age of patients and with regional prescription data to estimate individual fluoroquinolone reporting rate of adverse events. RESULTS: During the study period, 10 011 reports were received by the system (a mean annual reporting rate of approximately 185 per million inhabitants): 1920 referred to systemic antimicrobials, of which 432 (22.5%) involved fluoroquinolones. Pefloxacin was associated with the highest reporting rate (982 reports/daily defined dose/1000 inhabitants/day), followed by moxifloxacin (356), rufloxacin (221) and lomefloxacin (196). The most frequently reported reactions to fluoroquinolones involved the skin, but their percentage (25%) was significantly lower (p < 0.01) than those of other systemic antimicrobials (58.5%), whereas the percentages of reactions involving the central nervous (12.2 vs 3.6%), musculoskeletal (14.7 vs 0.3%) and psychiatric systems (9.3 vs 1.8%) were significantly higher (p < 0.01). We found some significant differences in the safety profiles of individual fluoroquinolones: ciprofloxacin was more frequently associated with skin reactions (p < 0.01), levofloxacin and pefloxacin with musculoskeletal (p < 0.01), and rufloxacin with psychiatric disorders (p < 0.05). Levofloxacin was the fluoroquinolone associated with the highest rate of serious tendon disorders; phototoxic reactions were more frequent with lomefloxacin, and toxic epidermal necrolysis and Stevens-Johnson syndrome were seen only with ciprofloxacin. CONCLUSIONS: The differences in the safety profiles should be taken into account when prescribing a fluoroquinolone to individual patients. PMID- 12534328 TI - The pharmacovigilance of pantoprazole: the results of postmarketing surveillance on 11 541 patients in England. AB - OBJECTIVE: To conduct a postmarketing surveillance study involving patients treated with pantoprazole in general practice in the 6-month period following the launch of this drug in November 1996. Prescription-event monitoring (PEM) provides data on a large cohort of patients in 'real life' settings. The aim was to monitor the safety of pantoprazole as used in general practice. It was the third proton pump inhibitor launched in the UK. METHODS: Patients were identified by data from dispensed prescriptions (FP10s) written by general practitioners (GPs) in England for pantoprazole. Green forms were posted to GPs approximately 6 months following the first prescription identified. GPs were asked to list events that occurred during and after treatment. The incidence density of each event was calculated, ranked and the difference between the incidence of each event in the first and subsequent months of exposure was tested by constructing confidence intervals. RESULTS: Data were collected for 11 541 patients. The major indications for treatment were oesophageal reflux (22.7%) and dyspepsia (16.9%). Of GPs expressing an opinion, 81.9% reported pantoprazole to be effective. GPs reported 107 events as adverse drug reactions. The most frequent reason given for stopping was diarrhoea (106 patients), which corresponded with the adverse event with the highest incidence density. CONCLUSION: This PEM study has defined the reported safety profile of pantoprazole as used in general practice in England. The commonest adverse events found in this study have already been reported in the Summary of Product Characteristics. PMID- 12534326 TI - Tolerability of the triptans: clinical implications. AB - The triptans represent a relatively new class of compounds effective in the treatment of migraine. The safety and tolerability of these drugs have been extensively investigated since the first triptan (sumatriptan) became commercially available. A report on a very large population of patients tested during clinical trials and in postmarketing studies, confirms that these drugs are safe and well tolerated when correctly used. Adverse events are frequently reported, but are usually mild and only a few patients discontinue therapy because of them. These adverse events include, in particular, the so-called 'triptan symptoms' (tingling, sensation of warmth, etc.). The exact mechanism of chest symptoms reported by 20% of patients with migraine treated with triptans remains unclear, but are exceptionally related to a cardiac mechanism. CNS adverse events (i.e. somnolence) are also reported, but it is a matter of debate whether they are related to the pharmacological properties (i.e. lipophilicity) of the drug or are symptoms of the disease itself. The potential risk for drug overuse must be taken into account when the triptans are given to patients with a high frequency of migraine attacks. Clinical interaction of triptans with other drugs metabolised in the liver may theoretically influence the incidence of adverse events, but there is little evidence to support this assumption. There is no evidence of a teratogenic risk of triptans in pregnant women taking these drugs. PMID- 12534329 TI - Hospital visits caused by adverse drug reactions: incidence and preventability assessed in French primary care/emergency departments. PMID- 12534330 TI - Management of vasodilatory shock: defining the role of arginine vasopressin. AB - The rationale for an arginine vasopressin (argipressin) infusion was put forward after it was discovered that patients in shock states might have an endogenous arginine vasopressin deficiency. Subsequently, several investigations impressively demonstrated that arginine vasopressin can successfully stabilise haemodynamics even in advanced vasodilatory shock. We report on physiological and pharmacological aspects of arginine vasopressin, and summarise current clinical knowledge on employing a continuous arginine vasopressin infusion in critically ill patients with catecholamine-resistant vasodilatory shock of different aetiologies. In view of presented experimental evidence and current clinical experience, a continuous arginine vasopressin infusion of approximately 2 to approximately 6 IU/h can be considered as a supplemental strategy to vasopressor catecholamines in order to preserve cardiocirculatory homeostasis in patients with advanced vasodilatory shock. Because data on adverse effects are still limited, arginine vasopressin should be reserved for patients in whom adequate haemodynamic stabilisation cannot be achieved with conventional vasopressor therapy or who have obvious adverse effects of catecholamines that result in further significant haemodynamic deterioration. For the same reasons, arginine vasopressin should not be used as a single, alternative vasopressor agent instead of catecholamine vasopressors. Future prospective studies will be necessary to define the exact role of arginine vasopressin in the therapy of vasodilatory shock. PMID- 12534331 TI - Treatment of lupus nephritis. AB - Renal involvement in systemic lupus erythematosus (SLE) is a serious complication of the disease. However, the prognosis of patients with lupus nephritis is continually improving with 10-year survival rates now greater than 75%. This improvement reflects earlier diagnosis due to more sensitive and specific diagnostic tests, better clinical appreciation of the natural history, and improved treatment of SLE and its manifestations. This review of the treatment of lupus nephritis has graded the level of evidence of specific treatment using the guidelines of the US Preventive Service Task Force. Although many new treatments have been advocated, the best evidence for treating proliferative lupus nephritis relies on a strategy combining specific treatment of the SLE as well as generalised treatment of the associated comorbidities. This strategy involves a combination of corticosteroids and cytotoxic agents plus or minus the adjunctive use of antimalarials, coordinated aggressive management of hypertension, proteinuria, infections, dyslipidaemia, thrombotic coagulopathy and potential renal replacement therapies. PMID- 12534332 TI - Management of chronic inflammatory demyelinating polyradiculoneuropathy. AB - This review briefly describes current concepts concerning the nosological status, pathogenesis and management of chronic inflammatory demyelinating polyradiculoneuropathy (CIDP). CIDP is an uncommon variable disorder of unknown but probably autoimmune aetiology. The commonest form of CIDP causes more or less symmetrical progressive or relapsing weakness affecting proximal and distal muscles. Against this background the review describes the short-term responses to corticosteroids, intravenous immunoglobulin (IVIg) and plasma exchange that have been confirmed in randomised trials. In the absence of better evidence about long term efficacy, corticosteroids or IVIg are usually favoured because of convenience. Benefit following introduction of azathioprine, cyclophosphamide, cyclosporin, other immunosuppressive agents, and interferon-beta and -alpha has been reported but randomised trials are needed to confirm these benefits. In patients with pure motor CIDP and multifocal motor neuropathy, corticosteroids may cause worsening and IVIg is more likely to be effective. General measures to rehabilitate patients and manage symptoms, including foot drop, weak hands, fatigue and pain, are important. PMID- 12534334 TI - Amoxicillin/clavulanic acid: a review of its use in the management of paediatric patients with acute otitis media. AB - Amoxicillin/clavulanic acid (Augmentin), Augmentin ES-600 is a well established, orally administered combination of amoxicillin (a semisynthetic antibacterial agent) and clavulanic acid (a beta-lactamase inhibitor). Amoxicillin/clavulanic acid shows good activity against the main pathogens associated with acute otitis media (AOM), including penicillin-susceptible and -intermediate strains of Streptococcus pneumoniae, and beta-lactamase producing strains of Haemophilus influenzae and Moraxella catarrhalis. It has moderate activity against penicillin resistant S. pneumoniae; a high-dose formulation has been developed with the aim of providing better coverage for penicillin-resistant strains. Amoxicillin/clavulanic acid (conventional formulations, mostly 40/10 mg/kg/day in three divided doses) produced clinical response rates similar to those of oral cephalosporin comparators and similar to or significantly greater than those for intramuscular ceftriaxone in randomised trials in paediatric patients with AOM (mean age approximately 2 to 5 years). Clinical response rates were generally similar for amoxicillin/clavulanic acid and macrolide comparators (mean patient age approximately 1 to 6 years), although significantly better clinical and bacteriological responses were seen versus azithromycin in one randomised trial (mean patient age approximately 1 year). The high-dose formulation of amoxicillin/clavulanic acid (90/6.4 mg/kg/day in two divided doses) eradicated a high proportion of penicillin-resistant S. pneumoniae (penicillin MICs 2 or 4 mg/L) in a large noncomparative trial in children with AOM (upper limit of the US indication for S. pneumoniae is 2 mg/L). Amoxicillin/clavulanic acid is generally well tolerated. A low total incidence of adverse events (3.6%) and no serious events were reported from a large paediatric postmarketing study. The most frequently reported adverse events in children are mild gastrointestinal disturbances. Diarrhoea is generally less frequent with twice-daily than with three-times-daily treatment. The new high-dose formulation showed similar tolerability to a conventional twice-daily formulation (45/6.4 mg/kg/day) in a well controlled trial. CONCLUSIONS: Amoxicillin/clavulanic acid is a well established broad-spectrum antibacterial treatment which is effective and well tolerated in the treatment of AOM in paediatric patients. The high-dose combination should prove valuable in treating AOM caused by penicillin intermediate and -resistant S. pneumoniae (approved in the US for penicillin MIC < or =2 mg/L). Based on recent recommendations and the available data, high-dose amoxicillin/clavulanic acid can be considered a treatment of choice for recurrent or persistent paediatric AOM (after failure of amoxicillin alone) where involvement of resistant pathogens is suspected. PMID- 12534335 TI - Effects of rare earth compounds on growth and apoptosis of leukemic cell lines. AB - To explore a new agent for inhibiting leukemic cells, we investigated the effects of rare earth compounds (lanthanum chloride and cerium chloride) on the growth and apoptosis of HL-60 and NB4 cells. The growth of HL-60 and NB4 cells was tested by 3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide (MTT) colorimetric assay. The apoptosis was measured by light microscopy, flow cytometry, and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL) method. The effect of LaCl(3) on normal bone marrow hematopoietic progenitor cells was evaluated by colony-forming unit-granulocyte-macrophage (CFU-GM) assay. Under our experimental conditions, MTT assay showed that 48-h treatment with 1, 2, and 3 mM LaCl(3) or 48- and 72-h treatments with 1 mM LaCl(3) could significantly inhibit the growth of HL-60 cells. Treatment with 2 and 4 mM CeCl(3) for 72 h could significantly inhibit the growth of NB4 cells. Apoptosis could be detected on treatment with 2 mM LaCl(3) for 24 h in HL-60 cells by light microscopic morphology examination, flow cytometric analysis, and TUNEL method. Apoptosis could be also detected on treatment with 2 mM CeCl(3) for 72 h in NB4 cells. Treatment with 1 mM LaCl(3) could arrest the transitions from G0/G1 to S phase. The granulocyte-macrophage colony formation of normal bone marrow cells was not significantly inhibited at lower concentrations of LaCl(3) (0.5 to 2 mM). Our results indicate that at certain concentrations, the rare earth compounds may inhibit the growth of leukemic cells, induce them to apoptosis, and have no significant inhibitory effects on normal bone marrow hematopoietic progenitor cells (CFU-GM). The mechanism needs to be further investigated. PMID- 12534336 TI - Spontaneously beating neonatal rat heart myocyte culture-a model to characterize angiotensin II at(1) receptor autoantibodies in patients with preeclampsia. AB - This report focuses on angiotensin II AT(1) receptor autoantibodies (anti-AT(1) AABs) in preeclamptic women. An enzyme-linked immunosorbent assay was described. Biotinylated peptide was incubated with anti-AT(1)-AABs. Streptavidin-coated magnetic particles bind the protein-autoantibody complex. Detection of anti-AT(1) AABs was performed using anti-human IgG3 peroxidase-coupled antibody. The color reaction of tetramethylbenzidine solution was stopped by adding 0.5 M H(2)SO(4). Optical density was measured at 450 nm (620 nm reference filter). Seventy-nine percent of anti-AT(1)-AAB-positive patients (measured by bioassay) showed an increase in optical density (>145%). The same biotinylated peptide was successfully used for purification of 6/6 anti-AT(1)-AABs. Chronotropic effects of purified antibodies were registered on primary cultured neonatal rat cardiomyocytes with the computer imaging system IMAGOQUANT. Western blot of coimmunoprecipitation of angiotensin II AT(1) receptor shows one band (molecular weight >40.0 kDa) in potassium thiocyanate eluate. PMID- 12534333 TI - Amyotrophic lateral sclerosis: progress and prospects for treatment. AB - Fifteen years ago, a role for excitotoxic damage in the pathology of amyotrophic lateral sclerosis (ALS) was postulated. This stimulated the development of riluzole, the only available treatment for the disease. Since then, the identification of abnormal forms of superoxide dismutase as the genetic basis of certain familial forms of ALS has provided a huge impetus to the search for new effective treatments for this devastating disease. Transgenic mouse models have been developed expressing these aberrant mutants that develop a form of motor neurone disease the progress of which can be slowed by riluzole. Studies in these mice have provided evidence for a role for excitotoxic, apoptotic and oxidative processes in the development of pathology. The mice can be used for testing molecules targeting these processes as potential therapies, to allow the most promising to be evaluated in humans. Several such agents are currently in clinical trials. Many previous clinical trials in ALS were insufficiently powered to demonstrate any relevant effect on disease progression. This situation has been to some extent remedied in the more recent trials, which have recruited many hundreds of patients. However, with the exception of studies with riluzole, the results of these have been disappointing. In particular, a number of large trials with neurotrophic agents have revealed no evidence for efficacy. Nonetheless, the need for large multinational trials of long duration limits the number that can be carried out and makes important demands on investment. For this reason, surrogate markers that can be used for rapid screening in patients of potential treatments identified in the transgenic mice are urgently needed. PMID- 12534337 TI - Bc3h1 myogenic cells produce an infectious ecotropic murine leukemia virus. AB - cDNAs representing an endogenous C-type ecotropic murine leukemia virus were isolated from a cDNA library constructed to represent mRNAs present in BC3H1 myogenic cells but not in C2C12 myogenic cells. RNA blot hybridization analysis using the cDNA inserts as probes revealed that BC3H1 cells produce MuLV-related transcirpts of at least three different size classes. A polymerase chain reaction enhanced assay for reverse transcriptase activity revealed the presence of reverse transcriptase in a viral pellet from medium conditioned by BC3H1 cells. A fungizone enhanced assay for syncitium formation provided further evidence of ecotropic retroviral particle production. Exposure of 3T3 cells to medium conditioned by BC3H1 cells, using conditions that facilitate infection, resulted in infection of the 3T3 cells, as confirmed by the syncitium formation assay. We conclude that BC3H1 cells produce an infectious ecotropic murine leukemia virus. Whether or not this feature of BC3H1 cells contributes to their inability to express some muscle-specific genes or to carry out myotube formation is unknown. Investigators will want to take into account that BC3H1 cells are virus producers when planning experiments that involve coculture of BC3H1 with other cell types, BC3H1 conditioned medium, retrovirally mediated transfection into BC3H1 cells, or study of the mCAT-1 amino acid transporter (the viral receptor) in BC3H1 cells. BC3H1 cells and the virus they produce may be of interest to those studying retroviral genomes and products and their effects. PMID- 12534338 TI - Further characterization of BC3H1 myogenic cells reveals lack of p53 activity and underexpression of several p53 regulated and extracellular matrix-associated gene products. AB - To catalog factors that may contribute to the completion of myogenesis, we have been looking for molecular differences between BC3H1 and C2C12 cells. Cells of the BC3H1 tumor line, though myogenic, are nonfusing, and withdraw from the cell cycle only reversibly, whereas cells of the C2C12 line fuse, differentiate terminally, and express several muscle-specific gene products that BC3H1 cells do not. Relative to C2C12 cells, BC3H1 cells underaccumulated cyclin-dependent kinase inhibitor p21 and underaccumulated transcripts for p21, GADD45, CDO, decorin, osteopontin, H19, fibronectin, and thrombospondin-1 (tsp-1). Levels of accumulation of H19, tsp-1, and larger isoforms of fibronectin messenger ribonucleic acid (mRNA) were found to increase in response to expression of myogenic regulatory factors as shown by their accumulation in differentiated myogenically converted 10T1/2 cells but not in 10T1/2 fibroblasts. BC3H1s accumulated a temperature-insensitive, geldanamycin-sensitive, misfolded form of p53 incapable of transactivating a p53 responsive reporter, consistent with underexpression of p21, GADD45, and tsp-1. BC3H1 and C2C12 cells were similar with respect to upregulation of p27 protein, downregulation of mitogen-activated protein kinase phosphatase-1 (MKP-1) protein, upregulation of retinoblastoma (Rb) mRNA, and nuclear localization of hypophosphorylated Rb. Cells of both lines expressed the muscle-specific 1b isoform of MEF2D. Although nonfusing in the short term, after more than 18 d in differentiation medium, some cultures of BC3H1 cells formed viable multinucleated cells in which the nuclei did not reinitiate synthesis of DNA in response to serum. Our findings suggest participation of tsp-1 and specific isoforms of fibronectin in myogenesis and suggest additional avenues of research in myogenesis and oncogenesis. PMID- 12534339 TI - The isolation of an immortalized and tumorigenic cell line from p21WAF1 null mouse bladders. AB - Given a role for the deregulation of p21WAF1 in the progression of bladder tumors, we examined the growth of cultured urothelial cells from wild-type and p21WAF1 null bladders. Bladders were excised, minced from euthanized p21WAF1 and wild-type mice, treated overnight with dispase, and then placed into flasks coated with collagen type I in Dulbecco modified Eagle medium with 10% fetal calf serum. After an overnight incubation, the media was replaced with a serum-free media and a portion of explants were treated with 12-O-tetrade-canoylphorbol-13 acetate (TPA) on day 7 and continued for either 4 or 9 wk. The urothelial origin of any surviving epithelial cells was determined by reverse transcription polymerase chain reaction (RT-PCR) using uroplakin II-specific primers, and the expression of the cell cycle-related proteins, p16INK4 and p19ARF, was examined by semiquantitative RT-PCR and Western blotting. Isolated wild-type and serially passaged p21WAF1 null epithelial-like cells were then injected subcutaneously into nude mice. We found that phorbol ester treatment at two different concentrations significantly enhanced uroepithelial colony formation from isolated wild-type mouse bladder tissue. On the other hand, significantly fewer urothelial colonies were derived from p21WAF1 null bladder cells treated with phorbol ester. Although there was apparent senescence and cell death of epithelial foci and stromal cells in phorbol ester-treated and -untreated p21WAF1 null cultures, after 3 mo there was an apparent subpopulation of epitheloid cells that overgrew each flask. There was a significant decrease in the number of these serially passaged cells in the G1 phase of the cell cycle when compared with initial explant wild-type or p21WAF1 null cells. This subpopulation of epitheloid cells expressed the mouse uroplakin II gene, indicating a urothelial phenotype, but did not express either the p16INKa or p19ARF proteins, whereas p21WAF1 null bladders express both proteins. There was also a high level of expression of the p53 protein and a significant decrease in the expression of the p19ARF transcript in both p21WAF1 null bladder and p21WAF1 null cells. These p21WAF1 null cells could be easily passaged and when injected subcutaneously into nude mice, large tumors developed. Therefore, it appears that a subpopulation of urothelial cells from the p21WAF1 null bladder can develop a tumorigenic phenotype in vitro. PMID- 12534340 TI - Characterization and application of a vinblastine-selected CACO-2 cell line for evaluation of p-glycoprotein. AB - The role of the adenosine triphosphate-binding cassette (ABC) superfamily of membrane transporters is well documented in tumor cell multidrug resistance. More recently, growing evidence of their influence on oral bioavailability, drug excretion rates, and drug-drug interaction potential at the intestinal level has stimulated much investigation. Our laboratory is interested in evaluating the apical (AP) ABC transporter P-glycoprotein (Pgp [mdr-1]) for its role in xenobiotic efflux at the intestinal level. We propagated Caco-2 cells in the presence of vinblastine (a cytotoxic, Pgp substrate) to promote transporter expression though selection. That is, the cell population expressing Pgp, or with the capacity to up-regulate Pgp expression, survived and expanded in the presence of vinblastine. We have used this selected cell line (Caco-2 VinB) to develop a fluorescent-based assay to study the chemical modulators of Pgp activity. Using the Caco-2 VinB cells, we have successfully demonstrated the differential potency of previously characterized Pgp inhibitors. In addition, we conducted a morphological evaluation of the two cell lines using transmission, scanning, and confocal microscopy. Both cell strains differentiated into highly functional, polarized columnar epithelium, although the vinblastine-selected cell line had lost the phenotypic diversity observed in native Caco-2 populations. Increased Pgp expression was noted in Caco-2 VinB cells compared with the native cell line on Western blot analysis, which was localized to the AP surface using confocal microscopy and functionally demonstrated using transport assays. We believe that the Caco2 VinB cell line is a versatile tool for application in pharmaceutical drug development. PMID- 12534341 TI - Delayed dedifferentiation and retention of properties in dissociated adult skeletal muscle fibers in vitro. AB - Adult skeletal muscle fibers can be isolated and cultured but tend to dedifferentiate and sprout with time in culture. We examined isolated adult mouse flexor digitorum brevis muscle fibers under various culture conditions by monitoring maintenance of the same fibers at 2-d intervals using survival analysis. Fibers plated on laminin and cultured in serum-free media did not show sprouting and exhibited significantly (P < 0.0001) longer survival (median survival time, T(50) = 10.2 d) than fibers in serum-containing media (T(50) = 3.3 d). Cell proliferation was markedly suppressed in serum-free cultures. Multiple or delayed Ca(2+) transients in response to brief field stimulation were often observed in dedifferentiated fibers after several d in serum-containing media but were not observed in fibers in serum-free media. The addition of cytosine arabinoside to serum-containing cultures did not prolong fiber survival (P = 0.39) and did not eliminate sprouting but did greatly suppress proliferation of nonmuscle cells. Fibers cultured in agarose gel with serum exhibited small, bud like extensions but no sprouts and did not survive as long (T(50) = 6.2 d) as fibers plated on laminin and cultured in serum-free media (T(50) = 10.2 d) did. These results demonstrate that both morphological and physiological properties of fibers become modified in serum-containing media but can be retained by culturing without serum. PMID- 12534342 TI - In vitro cultivation of Wolbachia in insect and mammalian cell lines. AB - Wolbachia infecting the small brown planthopper, Laodelphax striatellus, were successfully maintained and cultivated in two insect and one mammalian cell lines. The bacteria with the planthopper ovary were introduced into the flasks with the cultures of the cell lines. The Wolbachia proliferated in mosquito (Aedes albopictus) and lepidopteran (Heliothis zea) cell lines and in the mouse cell line, L929. Proliferation of Wolbachia was confirmed by electron microscopy and quantitative polymerase chain reaction. This simple method for the cultivation of Wolbachia was applicable to other strains of Wolbachia, such as the one found in the lepidopteran eggs, and should facilitate fundamental and applied studies of this important group of microorganisms. PMID- 12534343 TI - Requirement for direct cross-talk between B1 and B2 kinin receptors for the proliferation of androgen-insensitive prostate cancer PC3 cells. AB - Stimulation of endogenous kinin receptors promotes growth of androgen-independent prostate cancer PC3 cells via activation of the mitogenic extracellular-signal regulated kinase (ERK) pathway. In the present study, we show that kinin-mediated mitogenic signalling and prostate-cell growth involves two subtypes of bradykinin (BK) receptors, B1R and B2R. Specific stimulation of either B1R or B2R by their respective agonists des-Arg(9)-BK and Lys-BK promoted ERK activation and cell growth, whereas selective blockade with specific antagonists des-Arg(9) [Leu(8)]BK and Hoe 140 respectively obliterated this effect, indicating the presence of both receptor subtypes. However, blockade of B1R also inhibited B2R mediated ERK activation and cell growth, and, similarly, antagonism of B2R inhibited the B1R-mediated response. Furthermore, both B1R and B2R agonists promoted internalization of B1R, whereas both receptor antagonists blocked this effect. The B1R ligands des-Arg(9)-BK and des-Arg(9)-[Leu(8)]BK had no effect on the binding of BK to B2R, as demonstrated by radioligand competitive binding studies. However, blockade of either B1R or B2R impaired the ability of the reciprocal receptor to produce inositol phosphates, suggesting that the interaction between B1R and B2R is proximal to activation of phospholipase C. These results provide evidence for the existence of B1R-B2R complexes in prostate cancer PC3 cells and demonstrate that antagonism of one receptor interferes with the signalling ability of the other, possibly at the level of receptor-Galpha(q) protein coupling. Selective inhibition of B1R, which is up-regulated in injured and cancerous tissue, may be beneficial for the treatment of advanced prostate cancer. PMID- 12534344 TI - A role for c-Jun N-terminal kinase 1 (JNK1), but not JNK2, in the beta-amyloid mediated stabilization of protein p53 and induction of the apoptotic cascade in cultured cortical neurons. AB - beta-Amyloid (A beta) peptide has been shown to induce neuronal apoptosis; however, the mechanisms underlying A beta-induced neuronal cell death remain to be fully elucidated. The stress-activated protein kinase, c-Jun N-terminal kinase (JNK), is activated in response to cellular stress and has been identified as a proximal mediator of cell death. In the present study, expression of active JNK was increased in the nucleus and cytoplasm of A beta-treated cells. Evaluation of the nature of the JNK isoforms activated by A beta revealed a transient increase in JNK1 activity that reached its peak at 1 h and a later activation (at 24 h) of JNK2. The tumour suppressor protein, p53, is a substrate for JNK and can serve as a signalling molecule in apoptosis. In cultured cortical neurons, we found that A beta increased p53 protein expression and phosphorylation of p53 at Ser(15). Thus it appears that A beta increases p53 expression via phosphorylation-mediated stabilization of the protein. Given the lack of availability of a JNK inhibitor that can distinguish between JNK1- and JNK2-mediated effects, we employed antisense technology to deplete cells of JNK1 or JNK2 selectively. Using this strategy, the respective roles of JNK1 and JNK2 on the A beta-mediated activation of the apoptotic cascade (i.e. p53 stabilization, caspase 3 activation and DNA fragmentation) were examined. The results obtained demonstrate a role for JNK1 in the A beta-induced stabilization of p53, activation of caspase 3 and DNA fragmentation. In contrast, depletion of JNK2 had no effect on the proclivity of A beta to activate capase 3 or induce DNA fragmentation. These results demonstrate a significant role for JNK1 in A beta-mediated induction of the apoptotic cascade in cultured cortical neurons. PMID- 12534345 TI - Physical interaction of tumour suppressor p53/p73 with CCAAT-binding transcription factor 2 (CTF2) and differential regulation of human high-mobility group 1 (HMG1) gene expression. AB - The CCAAT-binding transcription factor (CTF)/nuclear factor I (NF-I) group of cellular DNA-binding proteins recognizes the sequence GCCAAT and is implicated in eukaryotic transcription, as well as DNA replication. Molecular analysis of human CTF/NF-I cDNA clones revealed multiple mRNA species that contain alternative coding regions, apparently as a result of differential splicing. Expression and functional analysis established that individual gene products can bind to GCCAAT recognition sites and serve as both promoter-selective transcriptional activators and initiation factors for DNA replication. The interaction between CTF2 and p53/p73 was shown to modulate their ability to regulate transcription of their respective target genes. In the present paper, we report that p53 down-regulates the activity of the high mobility group 1 (HMG1) gene promoter, whereas p73alpha up-regulates the activity of this promoter. Furthermore, CTF2 transactivates p53 induced p21 promoter activity, but inhibits p73alpha-induced p21 promoter activity. Using deletion mutants, we found that the DNA-binding domains of both p53 and p73alpha are required for physical interaction with CTF2 via the regions between amino acid residues 161 and 223, and 228 and 312 respectively. CTF2 enhances the DNA-binding activity of p53 and inhibits the DNA-binding activity of p73alpha. These results provide novel information on the functional interplay between CTF2 and p53/p73 as important determinants of their function in cell proliferation, apoptosis, DNA repair and cisplatin resistance. PMID- 12534346 TI - The specificities of protein kinase inhibitors: an update. AB - We have previously examined the specificities of 28 commercially available compounds, reported to be relatively selective inhibitors of particular serine/threonine-specific protein kinases [Davies, Reddy, Caivano and Cohen (2000) Biochem. J. 351, 95-105]. In the present study, we have extended this analysis to a further 14 compounds. Of these, indirubin-3'-monoxime, SP 600125, KT 5823 and ML-9 were found to inhibit a number of protein kinases and conclusions drawn from their use in cell-based assays are likely to be erroneous. Kenpaullone, Alsterpaullone, Purvalanol, Roscovitine, pyrazolopyrimidine 1 (PP1), PP2 and ML-7 were more specific, but still inhibited two or more protein kinases with similar potency. Our results suggest that the combined use of Roscovitine and Kenpaullone may be useful for identifying substrates and physiological roles of cyclin-dependent protein kinases, whereas the combined use of Kenpaullone and LiCl may be useful for identifying substrates and physiological roles of glycogen synthase kinase 3. The combined use of SU 6656 and either PP1 or PP2 may be useful for identifying substrates of Src family members. Epigallocatechin 3 gallate, one of the main polyphenolic constituents of tea, inhibited two of the 28 protein kinases in the panel, dual-specificity, tyrosine-phosphorylated and regulated kinase 1A (DYRK1A; IC(50)=0.33 microM) and p38-regulated/activated kinase (PRAK; IC(50)=1.0 microM). PMID- 12534347 TI - The pro-urokinase plasminogen-activation system in the presence of serpin-type inhibitors and the urokinase receptor: rescue of activity through reciprocal pro enzyme activation. AB - The reciprocal pro-enzyme activation system of plasmin, urokinase-type plasminogen activator (uPA) and their respective zymogens is a potent mechanism in the generation of extracellular proteolytic activity. Plasminogen activator inhibitor type 1 (PAI-1) acts as a negative regulator. This system is complicated by a poorly understood intrinsic reactivity of the uPA pro-enzyme (pro-uPA) before proteolytic activation, directed against both plasminogen and PAI-1. We have studied the integrated activation mechanism under the repression of PAI-1 in a purified system. A covalent reaction between pro-uPA and PAI-1 was positively demonstrated but the reaction of PAI-1 with two-chain uPA was found to be at least 1000-fold faster. However, in spite of this very fast inhibition, two-chain uPA still became the dominant plasminogen activator when plasminogen was incubated with pro-uPA and PAI-1. The activity pattern observed under these conditions revealed an initial lag phase, followed by a continuous generation of minute amounts of active two-chain uPA, this uPA having a short lifetime before inhibition but still succeeding to generate new plasmin activity, thus preventing a complete inactivation of the feedback system. This property of the activation system was retained even in the simultaneous presence of PAI-1 and alpha(2) antiplasmin. Addition of soluble uPA receptor to the system did not change the role of pro-uPA and the same pattern was observed when pro-uPA was bound to the uPA receptor on U937 cells. The present mechanism maintains the system at standby level and may be triggered to increased activity without the need for an external initiating event. PMID- 12534348 TI - Protection from inactivation of the adenine nucleotide translocator during hypoglycaemia-induced apoptosis by mitochondrial phospholipid hydroperoxide glutathione peroxidase. AB - We demonstrated that mitochondrial phospholipid hydroperoxide glutathione peroxidase (PHGPx) first suppressed the dissociation of cytochrome c (cyt c) from cardiolipin (CL) in mitochondrial inner membranes and then apoptosis caused by the hypoglycaemia by the prevention of peroxidation of CL [Nomura, Imai, Koumura, Arai and Nakagawa (1999) J. Biol. Chem. 274, 29294-29302; Nomura, Imai, Koumura, Kobayashi and Nakagawa (2000) Biochem. J. 351, 183-193]. The present study shows the involvement of peroxidation of CL in the inactivation of adenine nucleotide translocator (ANT) and the opening of permeability transition pores by using the system of ANT-reconstituted liposome and isolated mitochondria. ANT activity appeared in dioleoyl phosphatidylcholine proteoliposome containing 10% (mol/mol) CL or phosphatidylglycerol (PG), but not other classes of phospholipids. ANT activity was competitively inhibited by the addition of cardiolipin hydroperoxide (CLOOH) in reconstituted liposomes containing CL. However, phosphatidylcholine hydroperoxide failed to inactivate the activity of ANT. The activity of ANT in reconstituted liposomes, including CLOOH, recovered when CLOOH in reconstituted liposome was reduced to hydroxycardiolipin by incubation with PHGPx. The activity of ANT was determined in rat basophil leukaemia RBL2H3 cells after their exposure to 2-deoxyglucose. ANT activity decreased to 50% of the control level by 4 h in response to apoptosis. In parallel, cyt c and apoptosis-inducing factor (AIF) were released from mitochondria. Suppression of the accumulation of CLOOH by overexpression of PHGPx in mitochondria effectively prevented the inactivation of ANT, the opening of permeability transition pores and the release of cyt c and AIF from mitochondria in hypoglycaemia-induced apoptotic cells. These findings suggest that mitochondrial PHGPx might be involved in the modulation of the activity of ANT and the opening of pores for the release of cyt c via the modulation of levels of CLOOH in the mitochondria. PMID- 12534350 TI - Autoimmune thyroid disease: propagation and progression. AB - Autoimmune thyroid disease is the archetype for organ-specific autoimmune disorders. Progress in treating these disorders lies in improvements of our understanding of the predisposing factors responsible, the mechanisms responsible for progression of disease, and the interaction between thyroid antigens and the immune system at the level of the T cell and antibody. In common with other autoimmune diseases, genetic, environmental and endogenous factors are required in an appropriate combination to initiate thyroid autoimmunity. At present the only genetic factors which have been confirmed lie in the HLA complex and CTLA-4 or a closely linked gene. Identifying other predisposing genes will require large scale family studies, or further insights into likely candidate genes. A number of environmental factors are known to predispose to autoimmune thyroid disease, including smoking, stress and iodine intake, while immunomodulatory treatments are revealing new pathways for disease emergence. The thyroid cell itself appears to play a major role in disease progression, interacting with the immune system through expression of a number of immunologically active molecules including HLA class I and II, adhesion molecules, cytokines, CD40 and complement regulatory proteins. New techniques, in particular phage display libraries, are providing the methods with which to identify autoantibody diversity in autoimmune thyroid disease and to provide tools for mapping autoantigenic epitopes. Application of these techniques is likely to lead to an understanding of how TSH receptor antibodies interact with the receptor to cause Graves' disease and also to the identification of novel orbital autoantigens in thyroid-associated ophthalmopathy. PMID- 12534349 TI - Epidermal-growth-factor receptor and metalloproteinases mediate thromboxane A2 dependent extracellular-signal-regulated kinase activation. AB - The signalling pathways that link G-protein-coupled receptors to mitogen activated protein kinases involve receptor and non-receptor tyrosine kinases and protein kinase C (PKC). We explored the pathways that are implicated in the thromboxane (TX) A(2)-dependent activation of extracellular-signal-regulated protein kinase (ERK) and the role of the two TX receptor (TP) isoforms, TP alpha and TP beta. ERK activation by IBOP, a TX analogue, was dependent on epidermal growth-factor receptor (EGFR) in TP alpha- or TP beta-transfected cells and in human aortic smooth muscle cells (hASMCs), since AG1478, a selective inhibitor of tyrosine phosphorylation of the EGFR, strongly blocked ERK and EGFR phosphorylation. In addition, EGFR transactivation leading to ERK activation involved matrix metalloproteinases (MMPs), since BB2516, an inhibitor of MMP, decreased ERK and EGFR phosphorylation in TP alpha- or TP beta-transfected cells. Moreover, we showed that both isoforms activate ERK phosphorylation in an Src kinase-dependent manner, whereas PKC was mainly implicated in ERK activation and EGFR phosphorylation by TP beta. In hASMCs, we showed that ERK activation depended on both pertussis-sensitive and -insensitive G alpha-proteins. We demonstrated further that EGFRs, PKC, Src kinase and MMPs are involved in ERK activation by TX. The results of the present study highlight a role for MMPs and PKC in EGFR transactivation triggered by the TPs and demonstrate this mechanism for the first time in primary cells, i.e. hASMCs. PMID- 12534351 TI - Leptin-induced weight loss is not solely mediated by anorexia. PMID- 12534352 TI - Association of cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) gene polymorphism and non-genetic factors with Graves' ophthalmopathy in European and Japanese populations. AB - OBJECTIVE: The development and severity of Graves' ophthalmopathy (GO) may result from a complex interplay of genetic and environmental factors. The aim of this study was to investigate the association of cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) gene polymorphism and non-genetic factors (age, sex, cigarette smoking) with GO in two different populations, Polish-Caucasians and Japanese. DESIGN: We investigated the distribution of CTLA-4 A49G polymorphism in 264 Caucasian patients with Graves' disease (GD), of which 95 had clinically evident GO (NOSPECS class >or=3) and 319 Japanese patients with GD, of which 99 had ophthalmopathy. The control groups consisted of healthy Polish adults (n=194), Polish centenarians (n=51) and Japanese adults (n=112). RESULTS: Allele G and G/G genotype were significantly increased in Caucasian patients with GD (48% and 25% respectively) and in Japanese patients with GD (69% and 47% respectively) compared with control groups. There were no significant differences in the G allele and G/G genotype frequencies in GO patients compared with GD patients without ophthalmopathy. Multiple logistic regression analysis demonstrated that cigarette smoking (P=0.03, odds ratio (OR)=1.7) and age of onset of GD over 42 Years (P=0.08; OR=1.6) were contributing factors associated with susceptibility to GO in Polish patients. In Japanese patients, a younger age of onset of GD had an effect on the development of GO (P=0.02, OR=1.8). CONCLUSIONS: (i) Allele G and G/G genotype confer genetic susceptibility to GD; (ii) CTLA-4 A49G polymorphism is not associated with the development of GO; (iii) different non genetic factors may contribute to GO in different populations. PMID- 12534353 TI - Serum thyroglobulin and 131I whole body scan after recombinant human TSH stimulation in the follow-up of low-risk patients with differentiated thyroid cancer. AB - OBJECTIVE: The 'standard' postoperative follow-up of patients with differentiated thyroid cancer (DTC) has been based upon serum thyroglobulin (Tg) measurement and (131)I whole body scan ((131)I-WBS) after thyroid hormone (T(4)) treatment withdrawal. However, (131)I-WBS sensitivity has been reported to be low. Thyroid hormone withdrawal, often associated with hypothyroidism-related side effects, may now be replaced by recombinant human thyroid stimulating hormone (rhTSH). The aim of our study was to evaluate the diagnostic accuracy of (131)I-WBS and serum Tg measurement obtained after rhTSH stimulation and of neck ultrasonography in the first follow-up of DTC patients. DESIGN: Ninety-nine consecutive patients previously treated with total thyroidectomy and (131)I ablation, with no uptake outside the thyroid bed on the post-ablative (131)I-WBS (low-risk patients) were enrolled. METHODS: Measurement of serum Tg and (131)I-WBS after rhTSH stimulation, and ultrasound examination (US) of the neck. RESULTS: rhTSH stimulated Tg was 1 ng/ml (Tg+) in 21 patients, including 6 patients with Tg levels >5 ng/ml. (131)I-WBS was negative for persistent or recurrent disease in all patients (i.e. sensitivity = 0%). US identified lymph-node metastases (confirmed at surgery) in 4/6 (67%) patients with stimulated Tg levels >5 ng/ml, in 2/15 (13%) with Tg >1<5 ng/ml, and in 2/78 (3%) who were Tg-negative. CONCLUSIONS: (i) diagnostic (131)I-WBS performed after rhTSH stimulation is useless in the first follow-up of DTC patients; (ii) US may identify lymph node metastases even in patients with low or undetectable serum Tg levels. PMID- 12534354 TI - A new missense mutation in the growth hormone-releasing hormone receptor gene in familial isolated GH deficiency. AB - OBJECTIVE: Mutations in the GH-releasing hormone (GHRH) receptor (GHRHR) gene (GHRHR) cause autosomal recessive familial isolated GH deficiency (IGHD). We searched for GHRHR mutations in two siblings with IGHD type IB and a history of parental consanguinity. DESIGN: We analyzed peripheral genomic DNA of an index patient. After identifying a novel mutation in the GHRHR, we performed functional studies in order to confirm that the mutation causes receptor malfunction. METHODS: The entire GHRHR was analyzed in the index case by denaturing gradient gel electrophoresis. Abnormally migrating bands were isolated and sequenced. The mutated area was then sequenced in all family members whose DNA was available. The newly found mutation was inserted into a GHRHR cDNA. Wild-type and mutant cDNAs were expressed into CHO cells and the cyclic AMP (cAMP) response to GHRH was measured. In order to determine whether the mutant receptor was properly expressed on the cell membrane surface, CHO cells were transfected with wild-type or mutant GHRHR cDNA containing a FLAG epitope tag in the extracellular N terminus. RESULTS: Both patients were homozygous for a new missense mutation in codon 176, corresponding to the second transmembrane domain of the receptor protein that replaces alanine with valine (A176V). The mother and three unaffected siblings were heterozygous for the mutation; DNA from the father was not available. Cells expressing the A176V receptor had a significantly reduced cAMP response to GHRH, despite appropriate expression on the cell surface. CONCLUSIONS: We describe two siblings with IGHD due to a new mutation in the GHRHR that disrupts GHRH signaling and leads to GHRH resistance. PMID- 12534355 TI - Twelve months of treatment with octreotide-LAR reduces joint thickness in acromegaly. AB - OBJECTIVE: To evaluate the role of age, gender, duration and control of acromegaly on the reversibility of arthropathy. PATIENTS AND DESIGN: 30 de novo patients with active acromegaly, 30 cured patients and 30 healthy subjects were studied in a tranverse and an open longitudinal study design. METHODS: Shoulder, wrist and knee thickening was measured by ultrasonography at study entry in all 90 subjects and after 12 Months of treatment with octreotide-LAR (OCT-LAR) at a dose of 10-40 mg every 28 days in the 30 de novo patients. RESULTS: Thickness at all joint sites was greater in the active than in the cured patients and controls (P<0.001), and was greater in the cured patients than in the controls (P<0.001). There was no gender difference, but joint thickness was less in the patients with disease duration >10 Years. Age significantly correlated with wrist (r=-0.55; P<0.001), right knee (r=-0.45; P=0.01), and left knee thickness (r=-0.42; P=0.02) in patients with active disease, and with wrist thickness (r=0.88; P<0.0001) in controls. Twelve Months of OCT-LAR treatment led to disease control in 18 patients (60%). There was a decrease in the thickness of the shoulder (15.1+/ 3.2%), wrist (20.5+/-3.1%), right knee (22.2+/-3.4%) and left knee (18.2+/-2.8%) in all patients but the reduction in joint thickness at all sites was greater in the patients with controlled disease after OCT-LAR treatment than in the uncontrolled patients (P<0.01). Shoulder and right knee thickening normalized in respectively 11 (61.1%) and 16 (88.9%) well-controlled patients. CONCLUSIONS: Growth hormone and insulin-like growth factor-I (IGF-I) suppression by 12 Months' OCT-LAR treatment is accompanied by a significant decrease in the thickness of both weight-bearing and non-weight-bearing joints (mainly in patients whose disease is controlled) regardless of disease duration. These findings suggest that tIssue hypertrophy in the context of the acromegalic arthropathy can be improved by suppressing IGF-I levels. PMID- 12534356 TI - Plasma chromogranin A in patients with sporadic gastro-entero-pancreatic neuroendocrine tumors or multiple endocrine neoplasia type 1. AB - OBJECTIVE: As circulating chromogranin A (CgA) has been claimed to be the best general neuroendocrine marker so far available, we evaluated the usefulness of CgA determination in the clinical assessment of patients with sporadic gastro entero-pancreatic neuroendocrine tumors (GEP NETs) or multiple endocrine neoplasia type 1 (MEN 1). DESIGN AND METHODS: Plasma CgA levels were measured using a commercial enzyme-linked immunosorbent assay in 61 patients with sporadic GEP NET and in 25 with MEN 1 including 16 with GEP NET. Controls were 50 healthy volunteers, 46 patients with pituitary adenoma and 35 patients with primary hyperparathyroidism. RESULTS: The cutoff value for CgA established in our healthy subjects (as mean+2 s.d.) was 20 U/l. CgA levels were above the normal range in 71/77 patients with sporadic or MEN 1-related GEP NETs (92%), in four out of nine MEN 1 patients without GEP NETs (44%), and only in 22/81 control patients with pituitary or parathyroid disease (27%). Furthermore, CgA levels of over 100 U/l occurred in 36/77 patients with GEP NETs (47%) and only in one patient with a non functioning pituitary adenoma. In the patients with GEP NETs, both tumor burden and secretory activity affected CgA levels, and successful surgical resection was associated with markedly decreased CgA values. CONCLUSIONS: Plasma CgA was confirmed to be a reliable marker for GEP NETs. Moreover, in MEN 1 patients the finding of very high CgA levels strongly suggests the presence of a GEP NET, as both primary hyperparathyroidism and pituitary adenomas rarely cause marked CgA increases. PMID- 12534357 TI - During a corticotropin-releasing hormone test in healthy subjects, administration of a beta-adrenergic antagonist induced secretion of cortisol and dehydroepiandrosterone sulfate and inhibited secretion of ACTH. AB - OBJECTIVE: In chronic inflammatory diseases, serum levels of dehydroepiandrosterone (DHEA) sulfate (DHEAS) are low. Interestingly, several non inflammatory diseases display similarly low levels of DHEAS which points to other inhibitory factors such as an activated sympathetic nervous system (SNS) (e.g. in patients with heart failure, fibromyalgia, or cancer cachexia). We aimed to identify the influence of the SNS tone on stimulated adrenal steroid secretion in 16 male and 12 female healthy subjects. METHODS: One group were given oral propranolol 2 h before a corticotropin-releasing hormone (CRH) test, and levels of adrenocorticotropin (ACTH), cortisol, 17-hydroxyprogesterone (17OHP), androstenedione, DHEA, and DHEAS were measured. RESULTS: Propranolol treatment decreased heart rate (by 20%), diastolic blood pressure (by 20%), and plasma ACTH, and increased serum cortisol, serum DHEAS, and the molar ratio of cortisol/17OHP, cortisol/DHEA, and DHEAS/DHEA similarly in female and male subjects. CONCLUSIONS: A beta-adrenergic influence seems to decrease CRH stimulated cortisol in relation to ACTH and 17OHP, and decreases DHEAS in relation to DHEA. Although other workers have found beta-adrenergic stimulation of steroid secretion in cultured adrenocortical cells, the overall systemic influence of the SNS via beta-adrenoceptors seems to inhibit adrenal steroids under unstimulated and stimulated conditions. Sympathetic hyperactivity may be a common denominator for low levels of DHEAS in inflammatory and non-inflammatory diseases. PMID- 12534358 TI - Do reproductive hormones modify insulin sensitivity and metabolism in older men? A randomized, placebo-controlled clinical trial of recombinant human chorionic gonadotropin. AB - OBJECTIVE: In order to assess the hormonal determinants of insulin sensitivity and related components of the metabolic syndrome, we evaluated the effect of subcutaneous recombinant human chorionic gonadotropin (r-hCG; Ovidrel) on insulin sensitivity, vascular reactivity, leptin, insulin-like growth factor-I (IGF-I) and lipids in ambulant, community dwelling men >60 Years of age with serum testosterone 80% reduction of the size of differentiated thyroid carcinomas. Histological analysis of tumor specimens showed extensive necrosis and inflammatory cell infiltrates. The combination of IL-2/HSV-TK plus ganciclovir was significantly more efficient than IL-2 alone in eradicating tumor masses. The bystander effect was also obtained in vivo. CONCLUSIONS: These findings demonstrate the feasibility and efficiency of a combined immunomodulating and suicide gene therapy approach for thyroid carcinomas. PMID- 12534362 TI - Nitric oxide decreases the production of inositol phosphates stimulated by angiotensin II and thyrotropin-releasing hormone in anterior pituitary cells. AB - OBJECTIVE: Nitric oxide (NO) affects the synthesis of several second messengers, such as cyclic nucleotides, arachidonic acid metabolites and the intracellular calcium concentration, involved in the anterior pituitary hormone release. The present study was performed to investigate the effect of NO on phosphoinositide metabolism. METHODS: The synthesis of inositol phosphates (IPs) was studied in primary cultures of anterior pituitary cells from Wistar male rats. IPs (mono, bis and tris phosphates) were determined by ionic exchange chromatography. RESULTS: Sodium nitroprusside and DETA NONOate (DETA/NO) significantly decreased IP synthesis and prolactin release stimulated by angiotensin II (AngII) and thyrotropin-releasing hormone (TRH). These effects were not observed with decayed DETA NONOate (unable to release NO). LY-83583, a guanylyl cyclase inhibitor, completely reversed the inhibitory effect of DETA/NO on AngII-induced IP production. However, BAY 41-2272, a novel stimulator of the soluble guanylyl cyclase, did not mimic the effect of NO donors. Likewise, neither 8-Bromine cyclic GMP (8-Br-cGMP), an analog of cGMP, nor Sp-8-pCPT-cGMPS triethylamine, a cGMP-dependent protein kinase (PKG) stimulator, decreased IP synthesis stimulated by AngII. In addition, Rp-8-pCPT-cGMPS triethylamine, a PKG inhibitor, did not block the effect of NO. The decrease of IPs induced by DETA/NO was fully reversed by guanosine 5'-O-(3-thiotriphosphate) tetralithium salt, a non-hydrolyzable analog of GTP. CONCLUSIONS: The present work indicated that NO decreases IP synthesis stimulated by Ang II and TRH in anterior pituitary cells by a soluble guanylyl cyclase/cGMP/PKG-independent pathway, and suggested that NO affects some regulatory factor located between the plasma membrane receptor and G-protein. PMID- 12534361 TI - Immunohistochemical detection of cell cycle regulators, Fhit protein and apoptotic cells in parathyroid lesions. AB - OBJECTIVE: The pathological distinction between parathyroid neoplasms and hyperplasias remains difficult. Changes in cell cycle control may lead to clonal proliferation and precede tumorigenesis. The parathyroid adenoma 1 oncogene, subsequently identified as the gene encoding cyclin D1, has been shown to be important to parathyroid tumour development. In addition to cell proliferation, the mechanisms of parathyroid cell turnover include apoptosis. The tumour suppressor activity of the fragile histidine triad gene (FHIT) is linked to its proapoptotic function and cell cycle control. We attempted to evaluate the cellular proliferative kinetics and apoptotic function of the parathyroid glands in patients with non-familial hyperparathyroidism (HPT). DESIGN: TIssue specimens were taken from 40 patients with primary HPT (17 adenomas, two carcinomas and 21 primary hyperplasias) and from 30 patients with secondary HPT. Normal glands served as controls. METHODS: In a standard immunohistochemical procedure, monoclonal antibodies to Ki-67 antigen and single-stranded DNA were applied to detect cycling and apoptotic cells respectively; polyclonal antibodies to cyclin D1 and Fhit protein were used. Immunostaining was estimated by image analysis and statistical analysis was subsequently performed. RESULTS: Significantly higher proliferative and apoptotic indexes were detected in the diseased glands in comparison with normal controls. In neoplastic and secondarily hyperplastic glands, apoptotic indexes were higher than in primarily hyperplastic glands; the difference between neoplastic and primarily hyperplastic glands was statistically significant (P=0.034). Cyclin D1 was overexpressed in a considerable proportion of tumours (68.4%). A reduction of Fhit protein immunoreactivity was selectively noticed in carcinomas. CONCLUSIONS: In primary hyperplasia, the remarkable proliferation of parathyroid glands may be due to the reduction of the apoptotic process. FHIT gene abnormalities are worthy of investigation in parathyroid carcinogenesis. PMID- 12534363 TI - A yeast-based functional assay for the detection of the mutant androgen receptor in prostate cancer. AB - OBJECTIVE: Mutations in the ligand-binding domain of the human androgen receptor (AR) figure among the ways used by prostate adenocarcinoma (PCa) cells to escape androgen dependence. These mutations may broaden the specificity and/or affinity of the AR to other hormones, resulting in inappropriate receptor activation and thus affecting the PCa response to physiological stimuli and hormonal therapies. DESIGN: In order to clarify the impact of these mutations on disease progression and treatment, we have developed a yeast-based functional assay that allows the detection of mutant ARs and the analysis of their transactivation capacities in response to different ligands. METHODS: AR cDNA was directly cloned into an expression vector in a yeast strain that carries a reporter gene (ADE2) linked to an androgen-dependent promoter. The expression of the ADE2 gene and consequently the yeast cell growth in a selective medium depleted in adenine depends on the specificity of the AR for the ligand added to the medium. RESULTS: By analysing the transactivation capacities of different AR molecules in response to a broad range of steroid and non-steroid ligands, we have demonstrated that this assay can discriminate among wild-type AR, T877A, C685Y and L701H mutant ARs and that at least 1% of mutant ARs could be detected when mutant and wild-type ARs were mixed at the cDNA level. CONCLUSIONS: The data presented here show that this simple AR assay is convenient for the routine detection of mutant ARs in PCa and is also suitable to evaluate the antagonist activities of anti-androgen molecules. PMID- 12534364 TI - Altered circulating hormone levels, endothelial function and vascular reactivity in the testicular feminised mouse. AB - OBJECTIVE: Testicular feminised (Tfm) mice express a non-functional androgen receptor, and also have reduced levels of circulating testosterone. Recent studies support a cardio-protective role for testosterone since it elicits systemic and pulmonary vasodilatation. The aim of the present study was to determine whether androgen insensitivity and hypotestosteronaemia in the Tfm mouse are associated with abnormal vascular reactivity or hormone status. METHODS: Adult male Tfm and littermate control mice were killed and the blood collected. Femoral (diameter range = 183-508 microm) and pulmonary (diameter range = 320-816 microm) arteries were dissected and loaded in either a wire or pressure myograph, at 100 mmHg or 17.5 mmHg respectively. Pharmacological assessment of the vasoreactivity to potassium chloride (KCl, 80 mmol/l) and either noradrenaline (NA, 1 nmol/l-100 micromol/l) and acetylcholine (ACh, 0.1 100 micromol/l) or testosterone (1 nmol/l-100 micromol/l) was then made. RESULTS: Tfm mice had reduced levels of testosterone (1.8+/-0.3 nmol/l) compared with controls (9.3+/-2.0 nmol/l, P<0.001) and elevated levels of cholesterol (3.6+0.1 mmol/l) compared with controls (3.2+0.1 mmol/l, P<0.05). Femoral arteries from Tfm mice exhibited reduced vasoconstriction to 80 mmol/l KCl (3.27+/-0.23 mN/mm) compared with vessels from controls (4.44+/-0.41 mN/mm, P<0.05), and reduced endothelial-dependent vasodilatation to 0.1-100 micromol/l ACh (23.3+/-3.6% relaxation) compared with vessels from controls (41.6+/-5.4% relaxation, P<0.05). Vasoconstriction to NA (1 nmol/l-100 micromol/l) and vasodilatation to testosterone were unaffected. CONCLUSIONS: Androgen receptor deficiency and hypotestosteronaemia in the Tfm mouse reduced endothelial function and impaired voltage-operated calcium channel activity, which may pre-dispose to cardiovascular disease. Testosterone-induced vasodilatation was unaffected, demonstrating no involvement of the androgen receptor in this response. PMID- 12534365 TI - 5-HT1 and 5-HT2 receptor activation reduces N-methyl-D-aspartate (NMDA) stimulated LH secretion in prepubertal male and female rats. AB - OBJECTIVE: Excitatory amino acids and serotonin are involved in the control of gonadotropin secretion. The actions of these neurotransmitters are interconnected and recently we have reported that 5-HT(1) and 5-HT(2) receptor agonists blunted (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-stimulated GH secretion in prepubertal rats. The present experiments were carried out to analyze the effects of activation of different 5-hydroxytryptamine (5-HT) receptor subtypes on gonadotropin secretion and their role in the N-methyl-d aspartate (NMDA)-stimulated LH release. DESIGN AND METHODS: We analyzed the gonadotropin secretion after manipulation of serotoninergic and aminoacidergic systems and their interactions in 5-, 16- and 23-day-old male and female rats. To this end, serum LH and FSH concentrations were measured in rats treated with 5 hydroxytryptophan methyl ester (5-HTP) (a precursor of 5-HT synthesis) plus Fluoxetine (Fx, a blocker of 5-HT reuptake), d,l-p-chlorophenyl-alanine methyl ester (PCPA, a blocker of 5-HT synthesis), R-(+)-8-hydroxydipropylaminotetralin hydrobromide (8-OH-DPAT, an agonist of 5-HT(1A) receptors), (+/-)-2,5-dimethoxy-4 iodoamphetamine hydrochloride (DOI) and alpha-methyl-5-hydroxytryptamine (alpha Me-5-HT, agonists of 5-HT(2) receptors), and 1-Phenylbiguanide (1-PHE an agonist of 5-HT(3) receptors). In addition, the effects of 8-OH-DPAT and DOI on NMDA stimulated LH secretion were analyzed. RESULTS: Neither the activation nor blockade of the serotoninergic system modified LH secretion. Basal gonadotropin secretion remained unchanged in 23-day-old male and female rats after activation of 5-HT(1A), 5-HT(2) and 5-HT(3) receptors. The stimulatory effect of NMDA on LH secretion was blocked in both sexes after activation of the serotoninergic system, through specific 5-HT(1) and 5-HT(2) receptor agonists. CONCLUSIONS: Activation of serotoninergic receptors decreased the stimulatory effect of NMDA on LH secretion in prepubertal male and female rats. PMID- 12534366 TI - Is programming of glucocorticoid receptor expression by prenatal dexamethasone in the rat secondary to metabolic derangement in adulthood? AB - OBJECTIVE: Glucocorticoids may contribute to the association between retarded growth in utero and insulin resistance in adulthood. Administration of dexamethasone (dex) to pregnant rats results in low birth weight offspring, which develop glucose intolerance, hyperinsulinaemia and hypercorticosteronaemia. This may be explained by tIssue-specific differences in expression of glucocorticoid receptors (GR) in adult offspring: GR is increased in visceral fat and liver, and decreased in hippocampus and soleus muscle. However, cause and effect between altered GR expression, hypercorticosteronaemia, and hyperinsulinaemia remains to be established. DESIGN AND METHODS: Rats were treated with dex (100 microg/kg per day) or saline during the third week of pregnancy. In 5-8-Month-old male offspring, GR expression in insulin target tIssues was quantified by RNase protection assay in rats that were adrenalectomised (ADX group), sham operated (SHAM group), or adrenalectomised with supra-physiological corticosterone replacement (CORT group) (n=7-8 per group), and in rats treated orally with vehicle, metformin (43 mg/kg per day) or rosiglitazone (1 mg/kg per day), after 3 weeks. RESULTS: Manipulation of corticosterone concentration did not affect GR mRNA in skeletal muscle or adipose. In liver, sham-operated animals showed lower GR mRNA, but there was no difference between adrenalectomised and hypercorticosteronaemic animals (SHAM 0.11+/-0.01 ratio to beta-actin, vs ADX 0.22+/-0.02, CORT 0.23+/-0.02, (values expressed as means+/-s.e.m.), P<0.001). Rosiglitazone reduced GR mRNA by approximately 30% in liver of dex- and saline treated offspring (P<0.05), but had no effect on GR in adipose and skeletal muscle. Metformin abolished the 38% up-regulation of liver GR mRNA induced by antenatal dex and also reduced GR mRNA preferentially in muscle of dex-treated animals (0.14+/-0.01 vs 0.10+/-0.01; P=0.03). CONCLUSIONS: We conclude that neither hypercorticosteronaemia nor hyperinsulinaemia are sufficient to cause the changes in GR expression in dex-programmed rats, implying that these changes may be primary in determining the programmed insulin resistant phenotype. Normalisation of GR expression by metformin may be important in the mode of action of this anti-diabetic agent and may be especially useful to reverse programmed up-regulation of GR. PMID- 12534367 TI - Differential production of adrenal steroids by purified cells of the human adrenal cortex is relative rather than absolute. AB - OBJECTIVES: The adrenal cortex produces aldosterone, cortisol and androgens in response to ACTH and angiotensin II. To define the differential response of morphologically distinct cells of the adrenal cortex, we examined the phenotypical and functional characteristics of human adrenocortical cells. RESULTS: Tumour growth factor-beta receptor-1 (TGFbeta-R1) and CYP-11 were found to be expressed predominantly in the zona fasciculata, whereas human leukocyte antigen (HLA-DR) and CYP-17 were localised to the zona reticularis. The angiotensin II receptor, AT-1, was found to be predominantly expressed in the zona glomerulosa. Adrenocortical cells, separated by density, yielded two distinct fractions which displayed differential growth patterns. Lipid-rich cells of fraction I expressed TGFbeta-R1 and produced significantly more cortisol relative to androstenedione than unseparated or fraction II cells, whereas lipid poor cells of fraction II expressed HLA-DR and produced more androstenedione relative to cortisol in the presence of ACTH. Aldosterone production by fraction II was significantly greater than fraction I or unseparated cells. TGFbeta-R1 positive fasciculata-type cells separated into fraction I and HLA-DR-positive cells consistent with reticularis cells separated into fraction II. Aldosterone producing cells indicative of glomerulosa cells separated into fraction II. CONCLUSIONS: Our findings are consistent with the concept that all adrenocortical cells are capable of producing a range of steroids, but the relative production of cortisol, androgen and aldosterone differs. PMID- 12534368 TI - Insulin can block apoptosis by decreasing oxidative stress via phosphatidylinositol 3-kinase- and extracellular signal-regulated protein kinase dependent signaling pathways in HepG2 cells. AB - OBJECTIVE: Insulin has well-known activities in controlling energy metabolism, cellular proliferation and biosynthesis of functional molecules to maintain a biological homeostasis. Recently, several studies have suggested that insulin may protect cells from apoptosis in different cell lines; however, little is known about the nature of its anti-apoptotic activity. In many clinical disorders, including type 2 diabetes mellitus, oxidative stress and the production of reactive oxygen species (ROS) is increased. With these facts as a background, we examined here whether insulin protects HepG2 cells from apoptosis by decreasing oxidative stress and, if so, which signaling steps are involved in this process. METHODS: Intracellular DNA content, the degree of nuclear condensation or poly(ADP-ribose) polymerase hydrolysis was measured to verify the occurrence of apoptotic events. Caspase-3 activity and ROS accumulation within cells were also measured. Western blot analysis was performed to identify signaling molecules activated in response to insulin. RESULTS: Serum starvation resulted in a marked accumulation of ROS, activation of caspase-3, and subsequent apoptotic cell death which were, in turn, markedly blocked by the addition of insulin. The anti apoptotic activity of insulin was sensitive to blockade of two different signaling steps, activations of phosphatidylinositol 3-kinase (PI3 kinase) and extracellular signal-regulated protein kinase (ERK). CONCLUSION: Insulin exerts an anti-apoptotic activity by suppressing the excessive accumulation of ROS within cells through signaling pathways including stimulation of PI3 kinase and ERK in HepG2 cells. PMID- 12534369 TI - High glucose and insulin in combination cause insulin receptor substrate-1 and -2 depletion and protein kinase B desensitisation in primary cultured rat adipocytes: possible implications for insulin resistance in type 2 diabetes. AB - OBJECTIVE: The purpose of this study was to investigate the cellular effects of long-term exposure to high insulin and glucose levels on glucose transport and insulin signalling proteins. DESIGN AND METHODS: Rat adipocytes were cultured for 24 h in different glucose concentrations with 10(4) microU/ml of insulin or without insulin. After washing, (125)I-insulin binding, basal and acutely insulin stimulated d-[(14)C]glucose uptake, and insulin signalling proteins and glucose transporter 4 (GLUT4) were assessed. RESULTS: High glucose (15 and 25 mmol/l) for 24 h induced a decrease in basal and insulin-stimulated glucose uptake compared with control cells incubated in low glucose (5 or 10 mmol/l). Twenty-four hours of insulin treatment decreased insulin binding capacity by approximately 40%, and shifted the dose-response curve for insulin's acute effect on glucose uptake 2- to 3-fold to the right. Twenty-four hours of insulin treatment reduced basal and insulin-stimulated glucose uptake only in the presence of high glucose (by approximately 30-50%). At high glucose, insulin receptor substrate-1 (IRS-1) expression was downregulated by approximately 20-50%, whereas IRS-2 was strongly upregulated by glucose levels of 10 mmol/l or more (by 100-400%). Insulin treatment amplified the suppression of IRS-1 when combined with high glucose and also IRS-2 expression was almost abolished. Twenty-four hours of treatment with high glucose or insulin, alone or in combination, shifted the dose-response curve for insulin's effect to acutely phosphorylate protein kinase B (PKB) to the right. Fifteen mmol/l glucose increased GLUT4 in cellular membranes (by approximately 140%) compared with 5 mmol/l but this was prevented by a high insulin concentration. CONCLUSIONS: Long-term exposure to high glucose per se decreases IRS-1 but increases IRS-2 content in rat adipocytes and it impairs glucose transport capacity. Treatment with high insulin downregulates insulin binding capacity and, when combined with high glucose, it produces a marked depletion of IRS-1 and -2 content together with an impaired sensitivity to insulin stimulation of PKB activity. These mechanisms may potentially contribute to insulin resistance in type 2 diabetes. PMID- 12534370 TI - Characterization of G alpha 13-dependent plasma membrane recruitment of p115RhoGEF. AB - The Ras homology (Rho) guanine nucleotide exchange factor (GEF), p115RhoGEF, provides a direct link between the G-protein alpha subunit, alpha(13), and the small GTPase Rho. In the present study, we demonstrate that activated mutants of alpha(13) or alpha(12), but not alpha(q), promote the redistribution of p115RhoGEF from the cytoplasm to the plasma membrane (PM). We also show that the PM translocation of p115RhoGEF is promoted by stimulation of thromboxane A(2) receptors. Furthermore, we define domains of p115RhoGEF required for its regulated PM recruitment. The RhoGEF RGS (regulators of G-protein signalling) domain of p115RhoGEF is required for PM recruitment, but it is not sufficient for strong alpha(13)-promoted PM recruitment, even though it strongly interacts with activated alpha(13). We also identify the pleckstrin homology domain as essential for alpha(13)-mediated PM recruitment. An amino acid substitution of lysine to proline at position 677 in the pleckstrin homology domain of p115RhoGEF inhibits Rho-mediated gene transcription, but this mutation does not affect alpha(13) mediated PM translocation of p115RhoGEF. The results suggest a mechanism whereby multiple signals contribute to regulated PM localization of p115RhoGEF. PMID- 12534371 TI - Interfacial kinetic analysis of the tumour suppressor phosphatase, PTEN: evidence for activation by anionic phospholipids. AB - We investigated the kinetic behaviour and substrate specificity of PTEN (phosphatase and tensin homologue deleted on chromosome 10) using unilamellar vesicles containing substrate lipids in a background of phosphatidylcholine. PTEN displays the characteristics expected of an interfacial enzyme, since the rate of enzyme activity is dependent on the surface concentration of the substrate lipids used (mol fraction), as well as the bulk concentration. Surface-dilution analysis revealed the catalytic efficiency of PTEN for PtdIns(3,4,5) P (3) to be 200-fold greater than for either PtdIns(3,4) P (2) or PtdIns(3,5) P (2), and 1000-fold greater than for PtdIns3 P. The interfacial K (m) value of PTEN for PtdIns(3,4,5) P (3) was very low, reflecting the small proportions of this lipid that are present in cellular membranes. The catalytic-centre activity ( k (cat)) for PtdIns(3,4,5) P (3) was at least 200-fold greater than that for the water-soluble substrate Ins(1,3,4,5) P (4). The preference for lipid substrates may result from an interfacial activation of the enzyme, rather than processive catalysis of vesicular substrates. Moreover, both PtdIns(4,5) P (2) and univalent salts stimulated the activity of PTEN for PtdIns(3,4,5) P (3), but profoundly inhibited activity against Ins(1,3,4,5) P (4). The stimulatory effect of PtdIns(4,5) P (2) was greater in magnitude and more potent in comparison with other anionic phospholipid species. A mutation in the lipid-binding C2 domain (M-CBR3) that is biologically inactive did not alter overall catalytic efficiency in this model, but decreased the efficiency of the interfacial binding step, demonstrating its importance in the catalytic mechanism of PTEN. PMID- 12534372 TI - Haematopoietic lineage cell-specific protein 1 (HS1) promotes actin-related protein (Arp) 2/3 complex-mediated actin polymerization. AB - HS1 (haematopoietic lineage cell-specific gene protein 1), a prominent substrate of intracellular protein tyrosine kinases in haematopoietic cells, is implicated in the immune response to extracellular stimuli and in cell differentiation induced by cytokines. Although HS1 contains a 37-amino acid tandem repeat motif and a C-terminal Src homology 3 domain and is closely related to the cortical actin-associated protein cortactin, it lacks the fourth repeat that has been shown to be essential for cortactin binding to filamentous actin (F-actin). In this study, we examined the possible role of HS1 in the regulation of the actin cytoskeleton. Immunofluorescent staining demonstrated that HS1 co-localizes in the cytoplasm of cells with actin-related protein (Arp) 2/3 complex, the primary component of the cellular machinery responsible for de novo actin assembly. Furthermore, recombinant HS1 binds directly to Arp2/3 complex with an equilibrium dissociation constant (K(d)) of 880 nM. Although HS1 is a modest F-actin-binding protein with a K(d) of 400 nM, it increases the rate of the actin assembly mediated by Arp2/3 complex, and promotes the formation of branched actin filaments induced by Arp2/3 complex and a constitutively activated peptide of N WASP (neural Wiskott-Aldrich syndrome protein). Our data suggest that HS1, like cortactin, plays an important role in the modulation of actin assembly. PMID- 12534374 TI - Open access endoscopy. PMID- 12534373 TI - L-serine in disease and development. AB - The amino acid L-serine, one of the so-called non-essential amino acids, plays a central role in cellular proliferation. L-Serine is the predominant source of one carbon groups for the de novo synthesis of purine nucleotides and deoxythymidine monophosphate. It has long been recognized that, in cell cultures, L-serine is a conditional essential amino acid, because it cannot be synthesized in sufficient quantities to meet the cellular demands for its utilization. In recent years, L serine and the products of its metabolism have been recognized not only to be essential for cell proliferation, but also to be necessary for specific functions in the central nervous system. The findings of altered levels of serine and glycine in patients with psychiatric disorders and the severe neurological abnormalities in patients with defects of L-serine synthesis underscore the importance of L-serine in brain development and function. This paper reviews these recent insights into the role of L-serine and the pathways of L-serine utilization in disease and during development, in particular of the central nervous system. PMID- 12534375 TI - What maketh a surgeon? PMID- 12534376 TI - Elective resection and anastomosis for colorectal cancer: a prospective audit of mortality and morbidity 1976-1998. AB - BACKGROUND: The purpose of this paper is to audit the morbidity and mortality for colorectal cancer after open elective resection and anastomosis by a single colorectal surgeon (MK), with reference to any differences between intraperitoneal (IPA) and extra-peritoneal anastomoses (EPA). METHODS: The prospective documentation of postoperative complications was compiled by the surgeon, charge nurses and a research assistant (PB). The operations were performed in three hospitals between 1976 and 1998. RESULTS: Some 1392 consecutive patients were treated electively by 1418 resections with anastomosis. There were 23 postoperative deaths (1.6%). Significant adverse events, which were potentially avoidable, occurred in 10 (43.5%) of the patients who died. The morbidity rate was 41.6%. Clinical anastomotic leaks occurred more frequently in EPA (27/581, 4.7%) than in IPA (2/827, 0.2%; P < 0.0001). Anastomotic leak caused the death of two patients (0.14%).Routine prophylactic anticoagulation did not decrease the incidence of pulmonary embolism. Significant thrombophlebitis at the intravenous cannula site occurred in 54 patients (3.8%), wound infection in 29 (2.1%), and postural peripheral nerve injury in the upper limbs occurred in 11 patients (0.8%). Thirty-eight patients (2.7%) were returned to the operating theatre for 42 unscheduled operations. CONCLUSION: Mortality associated with elective resection of colorectal cancer with anastomosis is principally related to age, cardio-vascular disease and avoidable adverse events. A wide range of complications may follow this type of surgery, especially after extra peritoneal operations. A classification of anastomotic leaks is suggested to assist in comparisons of this complication which remains a significant concern following extra peritoneal anastomoses. PMID- 12534377 TI - Open access endoscopy in rural and remote Western Australia: does it work? AB - BACKGROUND: Access to diagnostic endoscopy is limited in rural and remote Western Australia. Published reports suggest open access referrals may result in over servicing, this is reduced by adherence to the American Society for Gastrointestinal Endoscopy (ASGE) guidelines. The aim was to assess whether an outreach surgical service offering open access endoscopy to rural areas was being over utilized. METHODS: Prospective data collection from all patients undergoing upper and lower endoscopy procedures between January 1996 and June 2000 were included in the present study. Indications for referral between the general practitioners and the visiting surgeons were reviewed in patient records and assessed for compliance with the ASGE guidelines. The groups were analysed for appropriateness of referrals and frequency of positive pathology investigations. Records for all patients undergoing colonoscopy were reviewed to determine the reason and number of cancelled procedures. RESULTS: A total of 772 endoscopies were performed and 75% were booked as open access services. The referral rate for procedures was greater for general practitioners (583) compared to the visiting surgeons (189), the overall compliance rate for approved indications using the ASGE guidelines for both groups was 92%. There was no significant difference in pathology found between groups. CONCLUSION: The present study shows that an outreach rural surgical service programme in Western Australia offering open access endoscopy conforms to international guidelines and does not induce unnecessary procedures. Rural patients benefit from a personal cost savings and convenience. There is an associated reduction in government-assisted travel costs to larger centres as well as decreased waiting lists. PMID- 12534378 TI - Prospective randomized trial of pre-emptive analgesics following ambulatory inguinal hernia repair: intravenous ketorolac versus diclofenac suppository. AB - BACKGROUND: A pre-emptive non-steroidal anti-inflammatory drug is routinely given to patients undergoing ambulatory inguinal hernia repair. The present prospective randomized trial was undertaken to compare the efficacy of intravenous ketorolac and rectal diclofenac for ambulatory inguinal hernia repairs. METHODS: Between June 1999 and February 2001, a total of 108 patients who underwent ambulatory inguinal hernia repairs under general anaesthesia were recruited. Patients were randomized to receive either intravenous ketorolac 30 mg immediately prior to induction of general anaesthesia (n = 54) or rectal diclofenac 50 mg after signing consent at the Day Surgery Centre (n = 54). RESULTS: The demographic features, hernia types, anaesthetic time, dosage of anaesthetic medication and operative details of the two groups were comparable. There was no significant difference in total amount of analgesic consumption and linear analogue pain scores after operation. With regard to recovery variables, the respective times taken to regain ambulation and micturition were similar in both groups. CONCLUSION: Diclofenac suppository 50 mg and intravenous ketorolac 30 mg provided equivalent postoperative analgesia following ambulatory inguinal hernia repair under general anaesthesia. Diclofenac suppository is an economical alternative to intravenous ketorolac. In the interests of cost containment rectal diclofenac could be considered the non-steroidal anti-inflammatory drug of choice for pre emptive analgesia. PMID- 12534380 TI - Surgical management of ductal carcinoma in situ in Australia in 1995. AB - BACKGROUND: In the present paper we describe the presentation and management of ductal carcinoma in situ (DCIS) of the breast in women in Australia in 1995. This representative, national data set provides a historical comparator for studies examining DCIS management that follow. METHODS: Surgeons identified by population based cancer registries as having treated a new diagnosis of DCIS between 1 April and 30 September 1995 completed a questionnaire on the presentation and management of each case. RESULTS: Two hundred and five surgeons supplied treatment details on 418 DCIS tumours in 415 women. Half of all tumours were detected at Breast Screen clinics and a further 25% were detected at other mammography centres. Twenty-six percent of tumours were palpable at presentation, 33% were multifocal and 55% were high grade (including comedocarcinoma). Breast conserving therapy (BCT) rather than mastectomy was utilized in 260 (62%)of cases. Tumours that were of low grade, small in size and not multifocal were more likely to be treated by BCT. Surgeons seeing six or moreDCIS cases in the 6-month period were more likely to utilize BCT. Of the conservatively treated cases, 22% were referred for a radiation oncology consultation. The most common reasons for treating DCIS with mastectomy were that the tumour was too extensive or multifocal (63%), it extended to margins of the specimen (42%), or patient concerns about recurrence (34%). CONCLUSIONS: In 1995 the majority of DCIS was treated with breast conserving surgery alone. Surgeons treating more DCIS cases were more likely to perform conservative surgery than surgeons treating only one DCIS case in the study period. PMID- 12534381 TI - Wound blisters post hip surgery: a prospective trial comparing dressings. AB - INTRODUCTION: Wound blisters are formed in the epidermis adjacent to surgical incisions and are a significant cause of morbidity following hip surgery. This study was designed to compare two commonly used primary dressings, namely a nonadherent absorbable (NAA) dressing and paraffin tulle gras (PTG). Monitoring for the subsequent development of wound blisters in the epidermis adjacent to the surgical incision was then undertaken. METHODS: The present study was performed in the setting of a prospective, randomised trial of 50 consecutive patients undergoing hip surgery. It was assumed there would be no difference in the development of blisters between the dressing groups. The alternative hypothesis was that PTG would, because of its properties, provide less friction in the epidermis and contribute to fewer wound blisters. RESULTS: A statistically significant difference (P = 0.0028) between the two dressing groups was observed with the NAA group developing blisters in 17 (64%) patients and the PTG group in only two (8%) of the patients. CONCLUSION: The present study would suggest that PTG produces fewer wound blisters following hip surgery when compared to the NAA type as a primary dressing. PMID- 12534383 TI - Fasciotomy for chronic compartment syndrome in the lower limb. AB - BACKGROUND: The aim of this study was to determine the success of fasciotomy of lower limb compartments with elevated intra-compartment pressure. METHODS: The present report is a retrospective cohort study at No. 3 Combat Support Hospital involving a survey and case note review of patients who had undergone fasciotomy for compartment syndrome at least 6 months earlier (December 1997-December 2000). RESULTS: Fourteen patients met the inclusion criteria. Eleven reported success with complete resolution of symptoms and return to military fitness levels. Two reported no change and significant residual symptoms. One reported worse symptoms and one required re-operation with a successful result. CONCLUSION: Surgical decompression is the accepted treatment of compartment syndrome as proven by pressure manometry, after failure of conservative treatment. PMID- 12534384 TI - Ankle fractures: functional and lifestyle outcomes at 2 years. AB - BACKGROUND: Ankle fractures form a high proportion of the total number of fractures treated in New Zealand. International studies show that there are mixed functional outcomes with differing fracture types and subsequently differing lifestyle outcomes. METHODS: Fracture clinic records and orthopaedic admissions books for Wellington Public Hospital, Capital Coast Health, -Wellington, were retrospectively reviewed to gain a population of patients who sustained ankle fractures for the period January--December 1998. These patients were asked to fill in postal questionnaires detailing their current ankle function and lifestyle, two years after fracturing their ankle. The patients' radiographs were reviewed to classify the types of ankle fractures sustained. RESULTS: Of 141 patients that sustained ankle fractures, 74 were followed up 2 years after their ankle fracture. All fracture types averaged Olerud-Molander ankle scores of 71.1. Weber A fractures averaged ankle function scores of 90, Weber B fractures 80, and Weber C fractures 78. Four patients (5%) achieved 'poor' results, 12 (16%) patients achieved a 'fair' result, 30 (41%) patients gained a 'good' result, 27 (36%) patients attained 'excellent' results. Lifestyle outcomes were reflected in the patient's ankle function outcomes (P < 0.05). CONCLUSION: Patients who sustain ankle fractures can be expected to be still experiencing functional difficulties two years post-treatment. PMID- 12534385 TI - Cholecystectomy for gall bladder dyskinesia: symptom resolution and satisfaction in a rural surgical practice. AB - BACKGROUND: Gall bladder dyskinesia describes a clinical entity with symptoms of biliary colic but without objective evidence of gallstones. Recent work has suggested laparoscopic cholecystectomy as an effective treatment although this approach has not gained widespread acceptance in Australia. METHODS: At the sole nuclear medicine facility in Mildura, Victoria, a retrospective search of patients with gallbladder ejection fractions or = 3 mm with naproxen (25.3%) was significantly higher than that with etoricoxib (7.4%) or placebo (1.4%; P < 0.001); the results were similar for ulcers of > or = 5 mm. CONCLUSIONS: The reduced toxicity of etoricoxib (less faecal blood loss and fewer endoscopically detectable lesions) suggests that use of this drug will may be associated with a reduced incidence of gastrointestinal perforations, ulcers and bleeds. PMID- 12534405 TI - Effect of oral omeprazole in reducing re-bleeding in bleeding peptic ulcers: a prospective, double-blind, randomized, clinical trial. AB - BACKGROUND: Endoscopic therapies and continuous intravenous omeprazole can decrease the morbidity and duration of hospital stay of patients with high-risk peptic ulcer. AIM: To evaluate the role of oral omeprazole in high-risk bleeders. METHODS: After injection therapy of 160 patients with high-risk peptic ulcer, 80 received oral omeprazole and 80 received placebo, and all were followed up. RESULTS: One hundred and forty-nine patients (71 omeprazole and 78 placebo) completed the study. Eleven patients were excluded from the study. Thirty-seven (25%) patients had gastric ulcer and 112 (75%) had duodenal ulcer. Fifty-seven (38%) ulcers showed visible vessels, 80 (54%) showed oozing of blood and 12 (8%) showed a spurting artery. Only one patient died (placebo group). The mean hospital stays were 62.8 +/- 28.6 h and 75 +/- 39 h in the omeprazole and placebo groups, respectively (P = 0.032). The mean amounts of blood transfused were 1.13 +/- 1.36 and 1.68 +/- 1.68 bags in the omeprazole and placebo groups, respectively (P = 0.029). The re-bleeding rate was lower in the omeprazole group than in the placebo group (12 vs. 26, respectively; P = 0.022). CONCLUSION: Oral omeprazole is effective in decreasing the hospital stay, re-bleeding rate and the need for blood transfusion in high-risk ulcer bleeders treated with endoscopic injection therapy. PMID- 12534406 TI - Occurrence of peptic ulcer disease in connective tissue disease patients associated with xerostomia. AB - BACKGROUND: Saliva plays a role in mucosal protection and ulcer healing. AIM: : To study whether decreased salivary production leads to peptic ulcer disease in connective tissue disease patients associated with xerostomia. PATIENTS AND METHODS: Two hundred and two connective tissue disease patients (90 with xerostomia and 112 without xerostomia) were enrolled. Their demographic data and use of medications were recorded. Peptic ulcer disease was confirmed by endoscopy. The stimulated salivary output and secretory epidermal growth factor level were measured. RESULTS: Compared with non-xerostomic counterparts, xerostomic patients manifested a higher occurrence of peptic ulcer disease (31% vs. 12%, P = 0.001), lower stimulated salivary output (9.3 +/- 4.1 vs. 22.9 +/- 5.9 mL/15 min, P < 0.001) and lower stimulated salivary epidermal growth factor output (1.40 +/- 0.77 vs. 3.00 +/- 0.96 ng/min, P < 0.001). Multivariate analysis disclosed that an older age (> or = 60 years) (odds ratio, 4.71; P < 0.001), xerostomia with stimulated salivary output of < or =1 mL/min (odds ratio, 7.54; P = 0.014) and the use of non-steroidal anti-inflammatory drugs (odds ratio, 5.76; P = 0.031) were the risk factors leading to peptic ulcer disease. In addition, xerostomic connective tissue disease patients receiving non-steroidal anti inflammatory drugs manifested an extremely high risk of development of peptic ulcer disease (odds ratio, 19.78; P < 0.001). CONCLUSIONS: Ageing, the use of non steroidal anti-inflammatory drugs and poor salivary function are potential risk factors for the development of peptic ulcer disease in patients with connective tissue disease. If these xerostomic subjects consume non-steroidal anti inflammatory drugs, they will encounter an extremely high peptic ulcer disease risk. PMID- 12534407 TI - Cost-effectiveness in Canada of intravenous proton pump inhibitors for all patients presenting with acute upper gastrointestinal bleeding. AB - BACKGROUND: The administration of proton pump inhibitors intravenously after endoscopic treatment of peptic ulcers significantly reduces the recurrence of bleeding. AIM: To evaluate the incremental cost-effectiveness in Canada of intravenous proton pump inhibitor before endoscopic therapy to patients presenting with acute upper gastrointestinal bleeding, compared with endoscopic treatment alone. METHODS: From a third-party payer perspective, we modelled the costs and effectiveness over 60 days of the two approaches using decision analysis. The probabilities of various outcomes, such as re-bleeding and the need for surgery, were taken from the published literature. We included the costs of intravenous proton pump inhibitor, therapeutic endoscopy, surgical procedures and hospitalizations, all expressed in 2001 Canadian dollars. RESULTS: In a hypothetical cohort of 1000 patients, the intravenous proton pump inhibitor approach resulted in mean savings of 20,700 Canadian dollars with 37 re-bleeding episodes averted. The investigation of uncertainty resulted in a likelihood of intravenous proton pump inhibitor being cost-effective of at least 0.73. CONCLUSION: It is common in Canada to administer intravenous proton pump inhibitors to patients with upper gastrointestinal bleeding even before endoscopic confirmation of bleeding peptic ulcers. Our results suggest that this approach has a high likelihood of being cost-effective. PMID- 12534408 TI - Effective intra-oesophageal acid suppression in patients with gastro-oesophageal reflux disease: lansoprazole vs. pantoprazole. AB - BACKGROUND: : Effective intra-oesophageal acid suppression is an important therapeutic goal in complicated and atypical gastro-oesophageal reflux disease. AIM: : To compare the efficacy of lansoprazole and pantoprazole in normalizing oesophageal acid exposure. METHODS: : Fifty patients with complicated or atypical gastro-oesophageal reflux disease were randomly assigned to receive 30 mg lansoprazole (n = 26) or 40 mg pantoprazole (n = 24) once daily. Three to four weeks after the start of treatment, patients underwent 24-h oesophageal pH monitoring whilst on therapy. If the results were improved but still abnormal, the dosage was doubled and pH monitoring was repeated. If oesophageal acid exposure was not improved, the patient was shifted to the alternative drug regimen. RESULTS: : Oesophageal acid exposure was normalized in all 26 patients treated with lansoprazole (in 35% of cases with a double daily dosage), whereas in six of the 24 (25%) patients treated with pantoprazole it was neither normalized nor lowered (P = 0.008). Accordingly, the mean percentage acid reflux time was significantly lower for the lansoprazole group (2.1) than for the pantoprazole group (5.8) (P = 0.032). CONCLUSIONS: : Effective intra-oesophageal acid suppression can be accomplished more reliably with lansoprazole than with pantoprazole in patients with complicated and atypical gastro-oesophageal reflux disease. PMID- 12534409 TI - Baclofen decreases acid and non-acid post-prandial gastro-oesophageal reflux measured by combined multichannel intraluminal impedance and pH. AB - BACKGROUND: Omeprazole controls acid but not non-acid reflux. The GABA B agonist baclofen decreases acid reflux through the inhibition of transient lower oesophageal sphincter relaxations (TLESRs) and should similarly decrease non-acid reflux. Using combined multichannel intraluminal impedance and pH (MII/pH), we compared acid and non-acid reflux after placebo and baclofen. METHODS: Nine healthy volunteers and nine heartburn patients underwent two 2-h studies of combined MII/pH in right lateral decubitus after a refluxogenic meal in random order: on placebo and after baclofen 40 mg p.o. Tracings were analysed for acid and non-acid reflux episodes, re-reflux and symptoms in the heartburn patients. RESULTS: In normal subjects baclofen significantly reduced the median number of episodes of acid (7 vs. 1, P = 0.02), non-acid (2 vs. 0, P = 0.005), and all reflux combined (10 vs. 2, P = 0.006); re-reflux was not reduced (0 vs. 0, P = N.S.). In heartburn patients, baclofen significantly decreased the median number of episodes of acid (15 vs. 6, P = 0.004), non-acid (4 vs. 2, P = 0.003), re reflux (2 vs. 0, P = 0.02), and all reflux combined (23 vs. 8, P = 0.004); it also reduced the median number of acid-related (9 vs. 1, P = 0.008) and non-acid related (1 vs. 0, P = 0.04) symptoms. CONCLUSIONS: Baclofen reduces post-prandial acid and non-acid reflux and their associated symptoms. GABA B agonists may have a role in treating GERD. PMID- 12534410 TI - A rapid-release 50-mg tablet-based 13C-urea breath test for the diagnosis of Helicobacter pylori infection. AB - BACKGROUND: Recently, a rapid-release 100-mg 13C-urea tablet with citrate supplement (Diabact UBT) showed excellent performance in a European population. AIM: To investigate the accuracy of a 50-mg tablet-based 13C-urea breath test protocol. METHODS: : Consecutive dyspeptic patients referred for upper endoscopy were recruited. 13C-Urea breath test was performed using a 50-mg 13C-urea tablet (Diabact UBT) and compared with the gold standard (rapid urease test and histology). Baseline, 10-min, 20-min and 30-min breath samples were collected in all cases. The cut-off values at each measurement interval were determined by three standard deviations above the mean excess delta 13CO2 excretion of Helicobacter pylori-negative patients. RESULTS: Two hundred patients (150 before therapy and 50 after therapy) were available for analysis, with a mean age of 48.4 years, and 99 patients (50%) were H. pylori positive. The sensitivity and specificity of the 50-mg tablet-based 13C-urea breath test at 10 min, 20 min and 30 min were 100% and 98%, 100% and 100%, and 100% and 98%, respectively. CONCLUSION: A 20-min, 50-mg tablet-based 13C-urea breath test (Diabact UBT) protocol is highly accurate for the diagnosis of H. pylori infection. PMID- 12534411 TI - Effect of different proton pump inhibitors, differences in CYP2C19 genotype and antibiotic resistance on the eradication rate of Helicobacter pylori infection by a 1-week regimen of proton pump inhibitor, amoxicillin and clarithromycin. AB - AIM: To investigate the effect of different proton pump inhibitors, S-mephenytoin 4'-hydroxylase (CYP2C19) genotype and antibiotic susceptibility on the eradication rate of Helicobacter pylori. METHODS: One hundred and eighty-seven H. pylori-infected peptic ulcer patients were randomly treated with either rabeprazole (10 mg b.d.) or lansoprazole (30 mg b.d.) plus amoxicillin (750 mg b.d.) and clarithromycin (400 mg b.d.) for 1 week. The antibiotic susceptibility and CYP2C19 genotype (extensive or poor metabolizer) were investigated. RESULTS: The eradication rates in the rabeprazole-amoxicillin-clarithromycin (RAC) and lansoprazole-amoxicillin-clarithromycin (LAC) groups were 75% and 69%, respectively, on an intention-to-treat basis, and 80% and 75%, respectively, on a per protocol basis. The eradication rate for clarithromycin-resistant strains was significantly lower than that for clarithromycin-sensitive strains (24% vs. 86%, P < 0.05). For clarithromycin-sensitive strains in the LAC group, there was a tendency for a lower eradication rate in extensive than poor metabolizers. The eradication rate in extensive metabolizers in the RAC group tended to be higher than that in extensive metabolizers in the LAC group (89% vs. 78%, P = 0.079726). CONCLUSIONS: The success of the 1-week proton pump inhibitor-amoxicillin clarithromycin regimen depends on the susceptibility of H. pylori to clarithromycin. Moreover, differences in CYP2C19 genotype influence the eradication rates of lansoprazole-based therapy, and the rabeprazole-based regimen has an advantage especially in extensive metabolizers. PMID- 12534412 TI - Double-blind, randomized, placebo-controlled study of nitazoxanide in the treatment of fascioliasis in adults and children from northern Peru. AB - BACKGROUND: Human fascioliasis is a significant world-wide health problem, and massive or repeated infections by Fasciola hepatica can lead to considerable morbidity. AIM: : To evaluate the safety and efficacy of nitazoxanide, when compared with placebo, in the treatment of fascioliasis in adults and children from northern Peru. METHODS: A double-blind, placebo-controlled study was carried out in 50 adults and 50 children infected with F. hepatica. The diagnosis of infection was based on the presence of F. hepatica eggs in one stool sample obtained before inclusion in the study. Patients were randomized to receive treatment with either a 7-day course of nitazoxanide (100 mg b.d., age range 2-3 years; 200 mg b.d., age range 4-11 years; 500 mg b.d., age > 12 years) or matching placebo. Three post-treatment stool examinations were carried out between 30 and 90 days after initiation of treatment. RESULTS: The parasite was eliminated in 18 of 30 (60%) adults completing the study who received nitazoxanide vs. one of eight adults in the placebo group (P = 0.042), and similarly in 14 of 35 (40%) children completing the treatment vs. none of eight in the placebo group (P = 0.038). Only mild, transient adverse events were reported. CONCLUSIONS: A 7-day course of nitazoxanide was effective in adults and children in the treatment of F. hepatica, when compared with placebo. PMID- 12534413 TI - Study of liver function in healthy elderly subjects using the 13C-methacetin breath test. AB - BACKGROUND: The issue of whether or not liver function is compromised in the healthy elderly population remains unsolved. AIM: To investigate the putative age related modifications of hepatic function using the 13C-methacetin breath test. Because endogenous CO2 production changes with age, motor activity and nutrition, a different form of processing the results was investigated. PATIENTS AND METHODS: Twenty-nine elderly subjects (mean age, 79.8 +/- 7.9 years; female/male ratio, 17/12) and 28 adult subjects (mean age, 40.6 +/- 12.3 years; female/male ratio, 13/15) underwent 13C-methacetin breath test and trans-abdominal echosonography with Doppler pulsed wave analysis of the coeliac axis and portal vein. RESULTS: Although the 13CO2 peak occurred within 15-30 min in both elderly and adult subjects, it was significantly decreased in the former (30.66% +/- 9.2% vs. 38.33% +/- 6.05%; P < 0.001), as was the cumulative excretion (33.07% +/- 7.06% vs. 39.81% +/- 5.68%; P < 0.001). When correcting for the effects of CO2 excretion by age, the age-related modification of the cumulative dose became more evident (elderly group 30.15% +/- 6.46% vs. adult group 37.97% +/- 5.92%; P < 0.0001). The elderly group also showed an increase in the intra-hepatic resistance index using Doppler pulsed wave analysis, which inversely correlated with the results of the breath test. CONCLUSIONS: Hepatic function is not well preserved in healthy humans throughout life and may be due to an increase in vascular resistance. PMID- 12534414 TI - Lipid-induced colonic hypersensitivity in irritable bowel syndrome: the role of 5 HT3 receptors. AB - BACKGROUND: Irritable bowel syndrome patients demonstrate colonic hypersensitivity after duodenal lipid infusion. AIM: To investigate the role of 5 hydroxytryptamine-3 (5-HT3) receptors in this sensory component of the gastrocolonic response in irritable bowel syndrome. METHODS: Fifteen female patients with diarrhoea-predominant irritable bowel syndrome completed a trial with the 5-HT3 receptor antagonist alosetron (1 mg b.d.) or placebo (b.d.) over 15 days, followed by the alternative treatment. Each treatment period was followed by a colonic distension trial before and after duodenal lipids. Changes in colonic thresholds, tone and compliance and viscerosomatic referral pattern after lipids were compared between treatments. RESULTS: With placebo, the colonic thresholds after lipids were significantly reduced for all studied sensations, whereas, with alosetron, the thresholds were significantly reduced only for first sensation and discomfort, but not for gas and pain. The reductions in thresholds did not differ significantly between treatments, but the pain threshold after alosetron tended to be less reduced compared with placebo (P = 0.10). The effects of lipids on tone, compliance and viscerosomatic referral pattern were unaffected by alosetron relative to placebo. CONCLUSIONS: 5-HT3 receptor antagonism reduces the lipid-induced colonic hypersensitivity in irritable bowel syndrome. However, 5-HT3 receptors do not seem to be the principal mediator, but may be a cofactor for the exaggerated sensory component of the gastrocolonic response in irritable bowel syndrome. PMID- 12534415 TI - Retained common bile duct stones: a comparison between biliary stenting and complete clearance of stones by electrohydraulic lithotripsy. AB - BACKGROUND: There is some uncertainty as to whether high-risk patients with difficult common bile duct stones should be subjected to a further endoscopic procedure for the complete removal of stones by electrohydraulic lithotripsy or whether permanent biliary stenting should be performed. AIM: To compare the outcome of permanent biliary stenting with electrohydraulic lithotripsy in this group of patients. METHODS: In a prospective study, 36 patients with difficult common bile duct stones were investigated: 19 underwent double pigtail insertion (stent group), whereas 17 underwent complete clearance of stones (electrohydraulic lithotripsy). RESULTS: In the electrohydraulic lithotripsy group, successful stone clearance was achieved in 76.5%, whereas, in the stent group, the success of stenting was 94.7%. A significant difference was detected in the actuarial incidence of recurrent acute cholangitis when the electrohydraulic lithotripsy group was compared with the stent group [one patient (7.7%) vs. 12 patients (63.2%), respectively; P = 0.002, log rank test]. A significant difference was detected in the actuarial frequency of mortality between the electrohydraulic lithotripsy and stent groups [seven patients (41.2%) vs. 14 patients (73.7%), respectively; P = 0.01, log rank test]. CONCLUSIONS: The removal of difficult common bile duct stones by electrohydraulic lithotripsy and further endoscopic retrograde cholangiopancreatography has a high success rate and a low complication rate even in the elderly. PMID- 12534416 TI - "Ask the physician": the modern relevance of an 18th century public health crisis. PMID- 12534417 TI - Cannabis and the evidence that led to its international control: a cautionary tale. PMID- 12534418 TI - Cannabis condemned: the proscription of Indian hemp. AB - AIMS: To find out how cannabis came to be subject to international narcotics legislation. METHOD: Examination of the records of the 1925 League of Nations' Second Opium Conference, of the 1894 Report of the Indian Hemp Drugs Commission and other contemporary documents. FINDINGS: Although cannabis (Indian hemp) was not on the agenda of the Second Opium Conference, a claim by the Egyptian delegation that it was as dangerous as opium, and should therefore be subject to the same international controls, was supported by several other countries. No formal evidence was produced and conference delegates had not been briefed about cannabis. The only objections came from Britain and other colonial powers. They did not dispute the claim that cannabis was comparable to opium, but they did want to avoid a commitment to eliminating its use in their Asian and African territories. PMID- 12534419 TI - Prison-based syringe exchange programmes: a review of international research and development. AB - Journal publications and conference presentations on prison-based syringe exchange (PSE) programmes were identified by a comprehensive search of electronic databases. Experts involved with development and evaluation of current PSE programmes or policy were contacted for reports, documents and unpublished material. Spanish information on PSE was translated for this review. We identified 14 papers specifically on PSE programmes in Switzerland (six papers), Germany (four) and Spain (four). The first PSE programme started in 1992 in Switzerland. As of December 2000, seven PSEs were operating in Switzerland, seven in Germany and five in Spain. There have been six evaluations of prison syringe exchange programmes and all have been favourable. Reports of drug use decreased or remained stable over time. Reports of syringe sharing declined dramatically. No new cases of HIV, hepatitis B or hepatitis C transmission were reported. The evaluations found no reports of serious unintended negative events, such as initiation of injection or of the use of needles as weapons. Staff attitudes were generally positive but response rates to these surveys varied. Overall, this review indicated that prison syringe exchange programmes are feasible and do provide benefit in the reduction of risk behaviour and the transmission of blood borne infection without any unintended negative consequences. PMID- 12534420 TI - Drug use patterns and infection with sexually transmissible agents among young adults in a high-risk neighbourhood in New York City. AB - AIMS: To determine relationships between drug use "hardness" (defined in increasing order of hardness as no drug use, marijuana use, non-injected heroin or cocaine use, crack smoking and injection drug use) and prevalences of several sexually transmissible infections among young adults in a high-risk neighbourhood. Drug users, particularly injection drug users and crack smokers, may be a core group for some sexually transmitted infections. DESIGN: Cross sectional survey and assays of young adults from (a) a household probability sample and (b) a targeted sample of youth who have used injected drugs, crack, other cocaine or heroin. SETTING: Bushwick, an impoverished New York City minority neighbourhood with major drug markets. PARTICIPANTS: A total of 363 18 24-year-olds from a household probability sample; 165 Bushwick 18-24-year-olds who have used injected drugs, crack, other cocaine or heroin. MEASUREMENTS: Drug use by self-report; serum- and urine-based assays for HIV, hepatitis B and C, syphilis, gonorrhoea, chlamydia and herpes simplex (type 2). FINDINGS: Household sample prevalences: HIV, hepatitis C and syphilis, 1%; gonorrhoea 3%; chlamydia 5%; past or present hepatitis B infection 8%; herpes simplex (type 2) 18%. In combined household and targeted samples, hepatitis C and HIV were concentrated among drug injectors. Herpes simplex (type 2), syphilis and hepatitis B increased among women with "hardest drug ever used". CONCLUSIONS: Using "harder" drugs is associated with some but not all of these infections. Prevention efforts should help youth avoid unsafe sex and higher-risk drugs. PMID- 12534421 TI - Psychometric properties of the Blood-borne Virus Transmission Risk Assessment Questionnaire (BBV-TRAQ). AB - AIMS: To develop a standard measure of blood-borne virus transmission risk behaviour, and examine the underlying psychometric properties. DESIGN: The Blood borne Virus Transmission Risk Assessment Questionnaire (BBV-TRAQ) was developed over three consecutive phases of the original BBV-TRAQ study in adherence to classical scale development procedures, culminating in the recruitment of a development sample of current injecting drug users via convenience and snowball sampling. SETTING: Needle and syringe programmes (NSPs), medical clinics, alcohol/drug agencies, peer-based and outreach organizations across inner and outer metropolitan Melbourne. PARTICIPANTS: Two hundred and nine current injecting drug users. The mean age was 27 years, 68% were male, 65% unemployed, 36% with prison history and 25% in methadone maintenance. MEASUREMENTS: BBV-TRAQ items cover specific injecting, sexual and skin penetration risk practices. BBV TRAQ characteristics were assessed via measures of internal and test-retest reliability; collateral validation; and principal components analyses. FINDINGS: The BBV-TRAQ has satisfactory psychometric properties. Internal (a=0.87), test retest (r=0.84) and inter-observer reliability results were high, suggesting that the instrument provides a reliable measure of BBV risk behaviour and is reliable over time and across interviewers. A principal components analysis with varimax rotation produced a parsimonious factor solution despite modest communality, and indicated that three factors (injecting, sex and skin penetration/hygiene risks) are required to describe BBV risk behaviour. CONCLUSIONS: The BBV-TRAQ is reliable and represents the first risk assessment tool to incorporate sufficient coverage of injecting, sex and other skin penetration risk practices to be considered truly content valid. The questionnaire is indicated for use in addictions research, clinical, peer education and BBV risk behaviour surveillance settings. PMID- 12534422 TI - Measuring therapeutic attitudes in the prison environment: development of the Prison Attitude to Drugs scale. AB - AIMS: To develop and test the validity of a scale measuring therapeutic attitudes among prison staff working with drug misusers. DESIGN: A cross-sectional postal questionnaire study using 27 statements with a five-point Likert scale. SETTING: Four prisons in the south-west of England PARTICIPANTS: A total of 252 prison staff (response rate 70%), including 67 for test-retest (response rate 57%). FINDINGS: The study resulted in a three-dimensional, nine-item scale: the Prison Attitude to Drugs scale (PAD). The three subscales measure confidence in skills (four items), personal rewards (three items) and job satisfaction (two items). Test-retest correlations for the questions were above 0.7, with each factor having an internal coherence (coefficient alpha) of greater than 0.7. CONCLUSIONS: The PAD is a reliable tool that can be used in the prison environment. PMID- 12534423 TI - Male drugs-related deaths in the fortnight after release from prison: Scotland, 1996-99. AB - AIMS: To assess if 15-35-year-old males released after 14 + days' imprisonment in Scotland, 1996-99, had a higher drugs-related death rate in 2 weeks after release than during subsequent 10 weeks; higher than expected death rate from other causes; and if drugs-related deaths in the first fortnight were three times as many as prison suicides. DESIGN: Confidential linkage of ex-prisoner database against deaths. SETTING: Scotland's male prisons and young offenders' institutions during July to December 1996-99; 19 486 index releases after 14+ days' incarceration. MEASUREMENTS: Relative risk of drugs-related death in the first 2 weeks after release (34 deaths) versus subsequent 10 weeks (23). Other causes of death (21) relative to expectation. Drugs-related deaths in first 2 weeks after release relative to suicides in prison (12). FINDINGS: Drugs-related mortality in 1996-99 was seven times higher (95% CI: 3.3-16.3) in the 2 weeks after release than at other times at liberty and 2.8 times higher than prison suicides (95% CI: 1.5-3.5) by males aged 15-35 years who had been incarcerated for 14+ days. We estimated one drugs-related death in the 2 weeks after release per 200 adult male injectors released from 14 + days' incarceration. Non-drugs related deaths in the 12 weeks after release were 4.9 times (95% CI: 2.8-7.0) the 4.3 deaths expected. CONCLUSION: Investment in, and evaluation of, prison-based interventions is needed to reduce substantially recently released drugs-related deaths. PMID- 12534424 TI - Concomitant drugs of misuse and drug using behaviours associated with fatal opiate-related poisonings in Sheffield, UK, 1997-2000. AB - AIMS: To examine the degree of involvement of concomitant drugs of misuse and other previously identified behavioural risk factors in acute accidental opiate related poisoning fatalities in Sheffield, 1997-2000. DESIGN: Retrospective analysis of coroners' records. SETTING: Sheffield, UK. PARTICIPANTS: All those who died from an acute accidental opiate-related poisoning in Sheffield between 1 January 1997 and 31 December 2000. MEASUREMENTS: Coronial data were collated under the headings: demographic characteristics, circumstances of death and toxicological findings. FINDINGS: Ninety-four deaths occurred over the study period. The majority of cases were regular users of illicit drugs. Approximately 20% of deaths were preceded by a period of abstinence from drug use, with imprisonment and hospitalization as the most common reasons. Sixty-one per cent of cases had concomitant drugs of misuse detected from toxicology most commonly benzodiazepines and/or alcohol. These were, however, found in relatively small concentrations and opiate blood concentrations were no lower in deaths where multiple substances were involved. Despite evidence to suggest that smoking is the preferred route of heroin administration in this region, the vast majority of cases involved injecting. CONCLUSIONS: Administration of an opiate via intravenous injection was the most consistent factor associated with these deaths over the period of this study. Co-administration of other central nervous system depressants, at least in lower quantities appear to be a feature rather than a risk factor per se in such fatalities. PMID- 12534425 TI - The prevalence and correlates of gamma-hydroxybutyrate (GHB) overdose among Australian users. AB - AIM: The aim of this study was to examine the correlates, context and risk perceptions regarding gamma-hydroxybutyrate (GHB) overdose among a sample of recreational GHB users in Australia. DESIGN: A cross-sectional survey of 76 GHB users who were administered a structured interview on GHB use. They were asked a series of questions regarding whether they had ever experienced a GHB overdose, the context of their most recent GHB overdose, and about their perceptions of the risks of GHB overdose. FINDINGS: This sample of GHB users had not had a long or extensive experience with GHB use; despite this, half (53%) had experienced a GHB overdose. This sample of GHB users appeared to be well-educated, employed and a history of either drug treatment or incarceration was uncommon. There were no differences between those who had or had not overdosed in terms of socio demographic characteristics, extent of other drug use or typical patterns of other drug use when using GHB. However, those who had overdosed on GHB had used it more times during their life-time, and had been using it for a longer period of time. CONCLUSIONS: GHB-related overdoses were common among a sample of GHB users who had only recently begun using the drug. The only apparent distinguishing factor between those who had and had not overdosed on GHB was the amount of experience with GHB use. PMID- 12534426 TI - A multilevel analysis of change in alcohol consumption in Quebec, 1993-98. AB - AIMS: Changes in drinking are subgroup-specific and vary by dimensions of drinking pattern. This study investigates changes in drinking patterns between 1993 and 1998 within various subgroups of Quebec society. DESIGN: A multilevel design was used in which respondents (level 1) were nested within demographic groups (level 2). PARTICIPANTS: Based on the 1993 and 1998 Quebec Health and Social Surveys, 40 598 respondents (level 1) were nested within 220 groups (level 2) based on age, SES status, gender and the year of survey. MEASUREMENTS: The effect of group characteristics were assessed for various dimensions of drinking pattern. FINDINGS: A significant part of the variance in drinking pattern is attributed to group membership (ranging from 8.6% to 18.3%). Except for the frequency of heavy drinking episodes, the data revealed a significant increase between 1993 and 1998 on all drinking pattern dimensions, which was more marked for higher SES groups for current drinking status. However, the frequency of heavy drinking decreased among the lower SES groups and remained stable among the higher SES groups. CONCLUSIONS: These results call for an increase in public health vigilance in monitoring trends in drinking patterns and their potential acute and chronic consequences. PMID- 12534427 TI - Prevalence and correlates of alcohol use: a population-based study in Hong Kong. AB - OBJECTIVE: To estimate the prevalence and pattern of alcohol use and to analyse the socio-demographic and biological correlates of alcohol drinking in Hong Kong Chinese. DESIGN: A population-based cross-sectional study conducted from December 1994 to October 1996. SETTING AND PARTICIPANTS: 2900 randomly selected subjects age 25-74 years who participated in the Cardiovascular Risk Factor Prevalence Study in Hong Kong. RESULTS: Alcohol consumers comprised 55.4% (95% CI: 52.8 58.0) of men and 19.4% (95% CI: 17.4-21.4) of women. The median weekly ethanol consumed by male and female drinkers were 9.6 g and 3.6 g, respectively. Beer was the main source of alcohol; 61.5% of drinkers consumed beer as their main drink. In stepwise multiple regression among drinkers, male sex, smoking, high density lipoprotein cholesterol, primary or below education, diastolic blood pressure and separated or widowed marital status were associated positively with weekly ethanol consumption. CONCLUSION: In this representative sample of Hong Kong adults, the majority were either non-drinkers or very light drinkers, which can be used as a benchmark to measure changes in drinking pattern in the future. The putative protective effect of alcohol on heart disease could be due to the higher level of HDL in moderate drinkers. PMID- 12534428 TI - Gender and ethnic differences in smoking, drinking and illicit drug use among American 8th, 10th and 12th grade students, 1976-2000. AB - AIMS: This paper examines ethnic differences in licit and illicit drug use among American 8th, 10th and 12th grade students, with a particular focus on girls. DESIGN: The study uses cross-sectional data from large, ethnically diverse, nationally representative samples of 8th, 10th and 12th grade girls. SETTING: Data were collected through questionnaires administered in schools. PARTICIPANTS: A total of 40 416 8th grade girls and 37 977 8th grade boys, 35 451 10th grade girls and 33 188 10th grade boys, and 33 588 12th grade girls and 31 014 12th grade boys took part in the study. FINDINGS: Across ethnic groups, drug use is highest among Native American girls and lowest among black and Asian American girls. Trend data suggest that there have been important changes in girls' drug use over time and that girls' and boys' drug use patterns are converging. CONCLUSIONS: Drug use is widespread among American adolescent girls. Future research should examine further girls' drug use and seek to identify whether risk and protective factors identified in past research, based on predominantly white samples, are also important predictors for drug use among non-white girls. PMID- 12534429 TI - A flicker paradigm for inducing change blindness reveals alcohol and cannabis information processing biases in social users. AB - AIM: To apply a new paradigm using transient changes to visual scenes to explore information processing biases relating to "social" levels of alcohol and cannabis use. PARTICIPANTS: Male and female student volunteers (n = 200) not self reporting substance-related problems. SETTING: Quiet testing areas throughout the university campus. DESIGN: A flicker paradigm, for inducing change blindness with lighter and heavier social users of alcohol (experiment 1, n= 100) and social users and non-users of cannabis (experiment 2, n= 100), explored the associations between habitual level of use and the latency to detection of a single substance related or neutral change made to a scene of grouped substance-related and neutral objects. MEASUREMENTS: Alcohol use was measured as the number of units of the heaviest drinking day from the previous week; cannabis use as the number of months of use in previous 12. Change-detection latency comparisons were used to evaluate processing biases. FINDINGS: In both experiments, (i) heavier social users detected substance-related changes quicker than lighter and non-users; (ii) lighter and non-users detected substance-neutral changes quicker than heavier users; (iii) heavier social users detected substance-related quicker than substance-neutral changes; and (iv) lighter and non-users detected substance neutral changes quicker than substance-related changes. CONCLUSIONS: Alcohol and cannabis processing biases are found at levels of social use, have the potential to influence future consumption and for this reason merit further research. PMID- 12534432 TI - Psychiatric research in an international perspective. The role of WPA. PMID- 12534433 TI - Antipsychotics and QT prolongation. AB - OBJECTIVE: To evaluate literature relating to cardiac QT prolongation and the use of antipsychotic drugs. METHOD: Literature searches of EMBASE, Medline, PsychLIT were performed in December 2001 and reference sections of retrieved papers scrutinized for further relevant reports. RESULTS: The Cardiac QTc interval is difficult to measure precisely or accurately but appears to be a useful predictor of risk of dysrhythmia (specifically torsade de pointes) and sudden death. It is less clear that drug-induced QTc prolongation gives rise to similar risks but data are emerging, linking antipsychotic use to increased cardiac mortality. Many antipsychotics have been clearly associated with QTc prolongation. Methodological considerations arguably preclude assuming that any antipsychotic is free of the risk of QTc prolongation and dysrhythmia. CONCLUSION: Available data do not allow assessment of relative or absolute risk of dysrhythmia or sudden death engendered by antipsychotics but caution is advised. Risk of dysrhythmia can very probably be reduced by careful prescribing of antipsychotics in low doses in simple drug regimens which avoid metabolic interactions. Electrocardiographic monitoring may also help to reduce risk but review by specialist cardiologist may be necessary. PMID- 12534434 TI - The importance of the QT interval: a review of the literature. AB - OBJECTIVE: QT interval (QTi) prolongation is generally associated with increased risk of ventricular arrhythmias such as torsade de pointes (TdP) and death. METHOD: Literature review based on publications identified by means of electronic and manual search. RESULTS: It has recently become apparent that not only antiarrhythmic drugs such as sotalol and quinidine, but also a variety of non antiarrhythmic drugs such as certain antihistamines, antimicrobial drugs, psychiatric drugs and cisapride, may have the ability to induce prolongation of the QTi and TdP. Special concern should be drawn to the coadministration of drugs that inhibit the metabolism of these drugs such as ketoconazole, itraconazol and erythomycin. Patients with congenital long QT syndrome, patients with heart disease, with hypokalemia or hypomagnesemia, and women have an increased risk. Every sign of dizziness or syncope should be regarded as a warning sign of possible arrhythmia in patients treated with drugs that potentially prolong the QTi. CONCLUSION: Measurement of the QTi before and during treatment is generally recommended in high-risk patients. PMID- 12534435 TI - Prevalence of mental disorders among 15-17-year-old male adolescent remand prisoners in Denmark. AB - OBJECTIVE: To determine the past year prevalence of mental disorders of 15-17 year-old adolescent remand prisoners in east Denmark. METHOD: One hundred 15-17 year-old boys from east Denmark consecutively remanded during 1 year were interviewed with diagnostic instruments to obtain ICD-10 diagnoses. All were screened in The Danish Psychiatric Case Register and The Danish Criminal Register. RESULTS: Past year prevalence of 'any mental disorder' was 69%, substance use disorders 41%. Two per cent had schizophrenia and 2% schizotypal disorder. Thirty-six per cent had 'probable personality disorder'. Conduct disorder was found in 31% and 1% had a hyperkinetic disorder. Ten per cent had previous registered psychiatric contact. CONCLUSION: The prevalence of mental disorders was found substantially higher compared with literature of population based samples. With an association between mental disorders and violence, early detection and treatment of mental disorders in adolescent delinquents is of importance in the prevention of violence. PMID- 12534436 TI - Taking cognizance of mental illness in schizophrenics and its association with crime and substance-related diagnoses. AB - OBJECTIVE: To analyse how committed crimes and substance-related diagnoses are associated with the age on the first contact with the psychiatric hospital system and the age at diagnosing of schizophrenia among schizophrenics. METHOD: In a register-based study including all Danes diagnosed with schizophrenia born after November 1, 1963, data on criminality, substance-related diagnoses and contacts with the psychiatric hospital system were analysed. RESULTS: Compared with the non-convicted schizophrenics the convicted were older on first contact with the psychiatric hospital system and older when the diagnosis of schizophrenia was first given. In contrast, having a substance-related diagnosis was associated with a younger age on first contact but did not influence the age at which the diagnosis of schizophrenia was given. CONCLUSION: It is important that both psychiatrists and the judicial system are aware of possible psychotic symptoms in criminal and abusing individuals to enable earlier detection and treatment. PMID- 12534437 TI - Do the Spontaneity and Anger and Aggression subscales of the Ward Atmosphere Scale form homogeneous dimensions? A cross-sectional study of 54 wards for psychotic patients. AB - OBJECTIVE: The main aim of this study was to examine the psychometric properties of the two subscales of the Ward Atmosphere Scale (WAS), Spontaneity and Anger and Aggression, which measure the level of expressed emotion. We wanted to examine if the items measuring attitude and behavior form different dimensions. METHOD: A total of 550 patients in 54 different psychiatric wards for psychotic patients completed the WAS and the Good Milieu Index. RESULTS: Removing, respectively, four items from the Spontaneity subscale, three items from the Anger and Aggression subscale the psychometric properties improved. The two revised subscales were named Spontaneous Behavior (SB) and Angry, Aggressive Behavior (AAB). A new 'attitude' subscale, comprising three items, was named Staff Attitude to Expressed Feelings (SAEF). CONCLUSION: For patients the attitude and behavior items seem to form separate dimensions. The three subscales examined are probably of central importance for patient satisfaction. PMID- 12534438 TI - Emotional reactivity to daily life stress in psychosis and affective disorder: an experience sampling study. AB - OBJECTIVE: To investigate the emotional reactivity to small disturbances in daily life in patients with non-affective psychosis (NAP), bipolar disorder (BD) and major depression [major depressive disorder (MDD)]. METHOD: Forty-two patients with NAP, 38 with BD, 46 with MDD, and 49 healthy controls were studied with the experience sampling method to assess (i) appraised subjective stress of small disturbances in daily life and (ii) emotional reactivity, reflected in changes in positive affect (PA) and negative affect (NA). RESULTS: Multilevel regression analyses showed an increase in NA in MDD, a decrease in PA in BD and both an increase in NA and a decrease in PA in NAP in association with the subjectively stressful situations, compared with the control subjects. CONCLUSION: Individuals with NAP, MDD and BD display differences in emotional stress reactivity. Type of mood disorder may exert a pathoplastic effect on emotional reactivity in individuals with MDD and BD. Individuals with NAP may be most vulnerable to the effects of daily life stress. PMID- 12534439 TI - The association between age and depression in the general population: a multivariate examination. AB - OBJECTIVE: In a large general population study we found a close to linear rise with age in the mean score and prevalence of self-reported symptoms of depression. The aim of this study was to examine if this linear relation prevailed when controlled for multiple variables and to examine factors that eventually explained the association. METHOD: Among individuals aged 20-89 years living in Nord-Trondelag County of Norway, 60 869 filled in valid ratings of the Hospital Anxiety and Depression Scale as well as many other variables. Covariates were grouped into a multivariate model with six blocks. Logistic regression was used to model the blocks and variables with caseness of depression as the dependent variable. RESULTS: The model explains a considerable part of the age related pattern on depression. The pattern became less distinct in the age groups above 50 years. Variables within the blocks of somatic diagnoses and symptoms, as well as impairment, had most explanatory power. CONCLUSION: Because of our large sample we were able to control for more relevant variables than earlier studies. In contrast to most other studies, we found that an age-related increase of the prevalence of depression persisted after control for multiple variables. PMID- 12534440 TI - Crazy? So what! Effects of a school project on students' attitudes towards people with schizophrenia. AB - OBJECTIVE: Aiming at promoting young people's mental health and reducing stigma towards people with schizophrenia, project weeks were carried out with secondary school students aged 14-18 years (n=90). Key to the project week is meeting a (young) person with schizophrenia. METHOD: Students' attitudes and behavioural intentions towards people with schizophrenia were assessed before and after the project. Parallelly, a control group of students were questioned (n=60). Assessment was repeated after 1 month. RESULTS: Despite expected ceiling effects, the project led to a significant reduction of negative stereotypes. For social distance, a positive trend could be observed. These developments were not present with the controls. Attitude changes were still evident at the 1-month follow-up. CONCLUSION: Results support the hypothesis that young people's attitudes about schizophrenia are susceptible to change. Antistigma projects at school level could thus be a promising approach to improving public attitudes and to preventing stereotypes from becoming reinforced. PMID- 12534441 TI - Change of immunoglobulins and complement factors in patients with self-injurious behaviour. AB - OBJECTIVE: As stress activates the inflammatory response system, and attempted suicide is connected with severe stress, we hypothesized that patients hospitalized for self-injurious behaviour have changed immunocompetence. METHOD: The concentration of immunoglobulins IgG, IgA, IgM, and the complement components C3 and C4 in 73 patients hospitalized for self-injurious behaviour was compared with those of 122 healthy controls. The immunoglobulins and complement were quantified by nephelometric technique. RESULTS: The levels of IgG and IgM were significantly lower, and the complement C3 and C4 were significantly higher in self-injurious patients compared with controls. This was valid in both genders and the effects did not interact with gender. CONCLUSIONS: This controlled study showed that the concentrations of immunoglobulins were reduced and complement components were increased in patients who are admitted to hospital for self injurious behaviour. PMID- 12534442 TI - Differences in fMRI and MRS in a monozygotic twin pair discordant for schizophrenia (case report). AB - OBJECTIVE: This paper presents functional magnetic resonance imaging (fMRI) and magnetic resonance spectroscopy (MRS) findings in a monozygotic twin pair discordant for schizophrenia. METHOD: Functional magnetic resonance imaging was used to determine hemispheric lateralization for speech and proton MRS (1H-MRS) was employed to assess the extent of putative insult to anterior hippocampus. RESULTS: Despite concordant right handedness, subject with schizophrenia displayed bilateral activation in areas subserving speech with greater extent of the total activated area compared with the healthy twin. The affected twin displayed relative bilateral decrease in N-acetylaspartate/creatin concentration in the anterior hippocampus compared with the healthy one. CONCLUSION: This is an evidence for non-genetic impairment of cerebral lateralization in monozygotic twin with schizophrenia. PMID- 12534444 TI - Methylenetetrahydrofolate reductase gene C677T mutation is related to the defects in the internal elastic lamina of the artery wall. AB - BACKGROUND: The C677T mutation of the methylenetetrahydrofolate reductase (MTHFR) gene leads to C/C, C/T and T/T genotypes, which affect the plasma homocysteine concentration in humans. In mini-pigs, high serum homocysteine levels are associated with defects in the internal elastic lamina (IEL) of the artery wall, which are apparently related to the migration of smooth muscle cells into the intima during atherogenesis. We studied the association between the MTHFR genotypes and the number of gaps in the IEL in the wall of the five major abdominal arteries. MATERIALS AND METHODS: The autopsy study included 123 subjects (90 males and 33 females) aged 18-93. For the light microscopy, a 0.5 cm circular segment of the coeliac, the superior mesenteric, the inferior mesenteric and the renal arteries were cut and embedded in paraffin blocks. The circumference of the IEL, the thickness of the intima and the number of the gaps per millimetre in the IEL were measured by MOP 3 image analysis. RESULTS: The T allele carriers (C/T and T/T) of the MTHFR gene had significantly less gaps in the IEL than the subjects with the C/C genotype in the superior mesenteric and in the left renal arteries (2.02 +/- 2.25 vs. 2.53 +/- 1.89, P < 0.04 and 0.56 +/- 1.09 vs. 1.82 +/- 2.66, P < 0.02, respectively). The trend was similar for the coeliac and the right renal arteries. CONCLUSIONS: Our result suggests that MTHFR polymorphism may be involved in the fragmentation of the IEL. PMID- 12534445 TI - Short-term cortisol infusion in the brachial artery, with and without inhibiting 11 beta-hydroxysteroid dehydrogenase, does not alter forearm vascular resistance in normotensive and hypertensive subjects. AB - BACKGROUND: Vascular tone is increased in primary hypertension, and glucocorticoids affect vascular tone. Local cortisol availability is modulated by activity of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD). As this activity may be decreased in patients with primary hypertension, vascular sensitivity to cortisol may be increased in these patients. We studied the acute effect of cortisol on forearm vascular resistance (FVR) by infusing cortisol directly into the brachial artery, both with and without inhibition of 11 beta-HSD, in normotensive and hypertensive subjects. DESIGN: Twenty normotensive volunteers and 20 patients with primary hypertension participated in the study. After a 10 min infusion of vehicle (glucose 5%), cortisol was infused into the brachial artery in three stepwise increasing doses (3.5, 10.5 and 35 microg per 100 mL of forearm volume), each for 10 min. Next, the participants received placebo or 500 mg glycyrrhetinic acid (GA) orally, and 150 min later the same infusion schedule was repeated. Forearm vascular resistance was measured during the last 5 min of the infused vehicle and of each dose. Arterial and forearm venous plasma samples for measurement of cortisol and cortisone were taken at the end of the infusions of glucose 5% and the highest cortisol dose. RESULTS: In both normotensive and hypertensive subjects, neither the infusion of cortisol nor the administration of GA changed FVR. Also 2 h after the cortisol infusion there remained no change in FVR in both the normotensive and hypertensive groups who received placebo. Following the infusion of the highest cortisol dose, total plasma cortisone levels in the venous plasma were decreased compared with levels in the arterial plasma (36 +/- 3 and 49 +/- 4 nmol L-1, respectively, P < 0.05). The protein bound venous cortisone was 37.1 +/- 4.8 nmol L-1 during the vehicle compared with 23.9 +/- 3.7 nmol L-1 during the cortisol infusion (P < 0.01), whereas the free cortisone level was not altered by the cortisol infusion. CONCLUSIONS: In both normotensive and hypertensive subjects, high-dose cortisol infusion both with and without 11 beta-HSD inhibition did not change FVR either immediately or after 2 h. We could not demonstrate in vivo 11 beta-HSD activity in the forearm vascular tissues. When binding of cortisone to CBG is changed, e.g. during cortisol infusion, arterio-venous changes in cortisone cannot reliably be used to assess (alterations in) local 11 beta-HSD activity. PMID- 12534446 TI - Effects of iloprost on adhesion molecules and F1 + 2 in peripheral ischemia. AB - BACKGROUND: Iloprost has beneficial effects on microcirculation by preventing platelet and leukocyte reciprocal activation, which is known to lead to endothelial damage and acute thrombosis. This drug also reduces inflammatory system activation by decreasing alpha M beta 2 integrin expression on the phagocyte membrane, might have a role in the protection and restoration of endothelial integrity and might interact with coagulation cascade activation. DESIGN: Forty patients were enrolled: 29 with systemic sclerosis (SSc) and 11 with peripheral artery disease (PAD). Iloprost was administered for 5 days in the first group and for 21 days in second group of patients. To ascertain whether iloprost modifies the parameters of endothelial and coagulation cascade activations, the plasma concentrations of S-ICAM-1 and F1 + 2 were detected in patients at baseline, after 5 days and, in PAD patients only, after 21 days of iloprost therapy. S-ICAM-1 is the endothelial counter receptor for alpha M beta 2 integrin and is a marker of endothelial cell activation; and F1 + 2 is a marker of coagulation cascade activation. RESULTS: After infusion of iloprost a significant decrease of S-ICAM-1 was observed in both the SSc (P < 0.002) and PAD patients (P < 0.004). Similarly, a significant decrease of F1 + 2 was observed in the SSc (P < 0.0004) and PAD patients (P < 0.003). CONCLUSIONS: The study provides evidence that iloprost reduces endothelial cells and coagulation cascade activations. Both of these mechanisms are responsible for improvement in microvascular functional capacity and for the long-term clinical benefit observed. After iloprost infusion, the SSc patients showed marked reductions in F1 + 2 and S-ICAM-1 concentrations that were statistically more significant relative to the PAD patients. PMID- 12534447 TI - Plasma vitamin E, total antioxidant status and vascular function in young adults. AB - BACKGROUND: Epidemiological studies have reported an inverse relationship between vitamin E status and coronary heart disease. This relationship has not, however, been confirmed by the majority of intervention studies, which have been carried out relatively late in the disease process. The protective effects of vitamin E may be more important earlier in life, before vascular changes have become established. This study investigated whether dietary vitamin E could prevent preclinical arterial changes in young adults relevant to the development of cardiovascular disease. MATERIALS AND METHODS: Measures of vascular function (arterial distensibility and endothelial-dependent and -independent vascular responses) were assessed by noninvasive high resolution ultrasound and related to plasma vitamin E and total antioxidant concentrations in 326 adults, aged 20-28 years. RESULTS: Neither vitamin E (alone or adjusted for lipids) nor total antioxidant status were significantly related to vascular endothelial function or arterial distensibility in either sex. There was no threshold level of vitamin E above which vascular function improved and neither vitamin E nor total antioxidant status interacted with any risk factor, such as smoking or increased low-density lipoprotein concentrations. CONCLUSIONS: Neither plasma vitamin E concentrations nor total antioxidant status achieved by dietary intake during young adulthood were related to vascular endothelial function or arterial distensibility. PMID- 12534448 TI - Prior aspirin use in unstable angina patients with modified plasma inflammatory markers and endothelial nitric oxide synthase in neutrophils. AB - BACKGROUND: Prior use of aspirin in patients with acute coronary syndrome has been associated with a lower incidence of acute myocardial infarction. The aim of this study was to explore if prior aspirin therapy in unstable angina (UA) patients could modify systemic inflammatory markers such as interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-alpha) and intercellular adhesion molecule-1 (ICAM-1) and the expression of endothelial nitric oxide synthase (eNOS) in neutrophils. MATERIALS AND METHODS: Unstable angina was defined as transient S-T segment changes without significant increases in CK and CK-MB. We studied 50 consecutive patients admitted to hospital within 24 h after the onset of chest pain. The number of patients with prior aspirin was significantly higher (n = 32) than those not taking aspirin (n = 18) on admission. RESULTS: Plasma levels of IL 6 and ICAM-1 were significantly increased in the UA patients when compared with the healthy control volunteers (n = 20) used as a reference for normal values. Plasma levels of both IL-6 and ICAM-1 were reduced in patients taking aspirin. There were no differences in the plasma levels of TNF-alpha between the UA patients and the control volunteers. The eNOS protein expression was also higher in neutrophils from the UA patients taking aspirin than in those not taking aspirin. CONCLUSION: Patients taking aspirin before UA showed a lower systemic inflammatory response and higher eNOS protein expression in their neutrophils PMID- 12534449 TI - Time-dependent effect of statins on platelet function in hypercholesterolaemia. AB - BACKGROUND: Reduction of platelet activity induced by statins has been described as a positive effect exerted by such molecules on vascular thrombotic events. However, the relations among cholesterol (LDL-C) reduction, the timing of the antiplatelet effect, the involved mechanisms and the doses of each statin able to reduce platelet function are not actually well known. The aim of our study was to evaluate the impact of simvastatin (20 mg day-1), atorvastatin (10 mg day-1), fluvastatin (40 mg day-1) and pravastatin (40 mg day-1) on platelet function in hypercholesterolaemic subjects with relation to (LDL-C), oxidized-LDL (ox-LDL) and antiport mechanism modifications. MATERIALS AND METHODS: Sixteen subjects were assigned to each treatment (40 males, 24 females, mean age 48.7 +/- 13.4, LDL-C 5.13 +/- 0,23 mmol L-1) and evaluated for platelet surface P-selectin (P sel), lipid profile, ox-LDL, platelet-associated ox-LDL (Pox-LDL), platelet cholesterol content, antiport mechanisms, and intracellular and systemic NO synthase every 7 days for one month. RESULTS: Our data show a strong relation between enhanced P-sel and Pox-LDL (r = 0.68, P < 0.01). Simvastatin, atorvastatin, fluvastatin and pravastatin reduce platelet activity after 1, 2, 3 and 4 weeks of treatment, respectively (P < 0.001, P < 0.001, P < 0.01, P < 0.05). Pox-LDL are modulated early by simvastatin, atorvastatin and fluvastatin Pox-LDL (r = 0.66, 0.65 and 0.52; P < 0.001, 0.001 and 0.01, respectively) whereas LDL-C and ox-LDL reductions associated to modifications of antiport activity act later. Moreover, they are the most relevant finding in pravastatin related subjects. CONCLUSIONS: Our data suggest a different impact of several statins on platelet function, which is initially related to interference with Pox LDL rather than LDL-C reduction. PMID- 12534450 TI - Is the gender difference in LDL size explained by the metabolic complications of visceral obesity? AB - BACKGROUND: Several studies have reported a significant gender difference in low density lipoprotein (LDL) size, with men being characterized by smaller, denser LDL particles than women, and it has been suggested that the contribution of the greater accumulation of visceral adipose tissue in men compared with women may be a factor potentially contributing to the gender difference in LDL heterogeneity. MATERIALS AND METHODS: We measured LDL particle size by 2-16% nondenaturing polyacrylamide gradient gel electrophoresis in 299 men and 231 women in whom visceral adipose tissue accumulation was measured by computed tomography. A fasting plasma lipoprotein-lipid profile was also obtained in all subjects. RESULTS: Overall, the men were characterized by a more deteriorated metabolic risk factor profile, which included higher plasma insulin and triglyceride levels, a greater visceral adipose tissue accumulation (P < 0.001) and smaller LDL particles (251.7 +/- 5.2 vs. 254.4 +/- 4.2 A, P < 0.0001). This gender difference in LDL peak particle diameter remained significant (252.4 +/- 4.3 vs. 253.5 +/- 4.3 A, P < 0.01) after adjustment for sex-specific differences in plasma triglyceride levels by covariance analysis. Significant negative correlations were noted between the LDL particle diameter and the triglyceride concentrations in both genders (r = -0.52 and r = -0.36, P < 0.0001 for the men and women, respectively), with no gender difference in this relationship being found. However, viscerally obese women (visceral adipose tissue levels > 100 cm2) with increased plasma triglyceride concentrations (> 2.0 mmol L-1) still had larger LDL particles than viscerally obese men with a similar elevation in their triglyceride levels (251.6 +/- 4.9 vs. 248.7 +/- 4.5 A, P < 0.01). CONCLUSIONS: Results of the present study suggest that the reduced LDL particle size observed in men compared with women cannot be entirely explained by their higher visceral adipose tissue accumulation and increased plasma triglyceride levels. Moreover, the gender difference in LDL size could be influenced, at least in part, by the severity of the hypertriglyceridaemic state. PMID- 12534451 TI - Effect of insulin on protein phosphatase 2A expression in muscle in type 2 diabetes. AB - BACKGROUND: Protein phosphatase 2A (PP2A) acts on a number of enzymes involved in the insulin regulation of glucose uptake and glycogen synthesis. This study was carried out to investigate the effect of insulin on PP2A expression in skeletal muscles of type 2 diabetic and control subjects. MATERIAL AND METHODS: Ten type 2 diabetic and 10 matched, control subjects were studied using the euglycaemic hyperinsulinaemic clamp technique combined with indirect calorimetry. Immunoreactive protein levels of the catalytic alpha subunit of PP2A (PP2A-C alpha) were measured in biopsies from the vastus lateralis muscle obtained in the basal and insulin-stimulated state. RESULTS: In type 2 diabetic subjects insulin mediated glucose disposal, glucose oxidation and nonoxidative glucose metabolism were reduced, whereas lipid oxidation was increased (all P < 0.05). Insulin down regulated PP2A-C alpha expression in skeletal muscle of the control subjects (P < 0.05) but not in the type 2 diabetic subjects. In the control subjects, the insulin-mediated decrease in PP2A-C alpha correlated with the insulin-mediated increase in glucose disposal, glucose oxidation, nonoxidative glucose metabolism (all P < 0.05) and decrease in lipid oxidation (P < 0.01). In the type 2 diabetic subjects these relationships were absent. CONCLUSIONS: Down-regulation of PP2A-C alpha expression by insulin in skeletal muscle seems to be associated with a normal insulin action on glucose storage, glucose and lipid oxidation. Impaired down-regulation of PP2A-C alpha expression by insulin may be a marker for insulin resistance and contribute to the pathogenesis of type 2 diabetes. PMID- 12534452 TI - Impact of diabetes duration and cardiovascular risk factors on mortality in type 2 diabetes: the Hoorn Study. AB - BACKGROUND: Several studies have reported differences in the mortality risk between diabetic subjects detected by screening and known diabetic patients. We studied mortality in relation to diabetes duration, and the contribution of other cardiovascular risk factors to the elevated risk. MATERIALS AND METHODS: Participants were type 2 diabetic subjects (n = 174) of a population-based cohort study. Of these, 95 were diagnosed by screening. Known diabetic subjects were grouped into two categories of diabetes duration, with a median duration of 2.4 and 11.2 years, respectively. We assessed the contribution of classical cardiovascular risk factors (dyslipidaemia, hypertension, and prior myocardial infarction), and of new cardiovascular risk factors (microalbuminuria, von Willebrand factor, sVCAM-1 and C-reactive protein) to the mortality risk during nearly 10 years of follow up. Cox's proportional hazards model was used to study the association of diabetes duration and mortality. RESULTS: The age- and sex adjusted relative risks of mortality were 2.06 (95% C.I. 1.04-4.10) and 3.19 (1.64-6.20) for the patients with short- and long-term diabetes compared with the screening-detected diabetic subjects, respectively. Adjustment for cardiovascular risk factors resulted in a reduction of mortality risk in both groups: 1.13 (0.51 2.50) and 2.39 (1.18-4.83), respectively. Mortality risk significantly increased with increasing diabetes duration, even after multiple adjustment (P-value for trend ranged from < 0.001-0.018). CONCLUSIONS: Mortality risk increased with increasing diabetes duration. In subjects with short diabetes duration the mortality risk could largely be attributed to other risk factors. In subjects with a longer diabetes duration, however, the elevated mortality risk was independent of these cardiovascular risk factors. PMID- 12534453 TI - Basal TSH levels compared with TRH-stimulated TSH levels to diagnose different degrees of TSH suppression: diagnostic and therapeutic impact of assay performance. AB - BACKGROUND: The estimated prevalence of endogenous subclinical hyperthyroidism varies from 4% to 6% and a basal thyroid stimulating hormone (TSH) level < 0.5 mU L-1 may be associated with increased mortality in subjects over 60 years of age who are not on thyroid medication. Exogenous TSH suppression is a mainstay in the treatment of thyroid cancer. Because of recent concerns about potential adverse effects, especially of endogenous TSH suppression on bone, the cardiovascular system and cognitive functions, subclinical hyperthyroidism obtained new clinical importance. We therefore re-evaluated the diagnostic value of basal and thyrotrop in TRH-stimulated serum TSH measurements using TSH assays with different sensitivities. MATERIALS AND METHODS: A total of 805 oral and nasal TRH stimulation tests were performed on 409 ambulatory subjects with low basal serum TSH concentrations of less than 0.1 mIU L-1. Basal serum TSH was measured either using a second generation assay (functional sensitivity > 0.03 mIU L-1) or two third generation assays (functional sensitivity 0.01 mIU L-1 and 0.007 mU L-1, respectively). Serum TSH concentration was determined before and 3 h after oral administration of 40 mg of TRH and before and 30 min after nasal administration of 2 mg of TRH. RESULTS: In the oral testing group, the basal TSH levels measured by the different TSH assays were 0.06 +/- 0.03, 0.04 +/- 0.02 and 0.03 +/- 0.02, respectively, whereas the peak TSH levels were 0.4 +/- 0.6, 0.4 +/- 0.6 and 0.3 +/- 0.5 in the patients with subclinical hyperthyroidism. In overt hyperthyroidism, the basal TSH levels were 0.06 +/- 0.02, 0.03 +/- 0.02 and 0.03 +/- 0.02, whereas the peak TSH levels were 0.19 +/- 0.3, 0.16 +/- 0.3 and 0.15 +/ 0.2, respectively. Basal TSH values could discriminate between different degrees of TSH suppression if measured with a third generation assay (P < 0.001), but not with a second generation assay. There was only a weak correlation between basal TSH and peak TSH when measured by a second generation assay (n = 126; r = 0.3; P < 0.001) in contrast to the strong correlation found using the third generation assays (n = 128; r = 0.7; P < 0.001 and n = 69; r = 0.8; P < 0.001, respectively). CONCLUSIONS: In view of the recent concerns about potential adverse effects in TSH suppression and based on our data, it is mandatory to select a TSH assay with a functional sensitivity of < or = 0.01 mIU L-1 for optimal titration of L-T4 suppressive therapy, especially in patients with thyroid cancer. If, however, only a second generation TSH assay is available, additional TRH testing allows a more careful titration of suppressive thyroxine therapy. PMID- 12534454 TI - The hormone-sensitive lipase i6 gene polymorphism and body fat accumulation. AB - BACKGROUND: The hormone sensitive lipase (HSL) catalyses the breakdown of adipose tissue triglycerides into free fatty acids. The objective of this study was to determine whether HSLi6 microsatellite allele 5 (A5) and/or homozygosity for this allele is associated with body fat in Swedes. DESIGN: A large case-control study on gender-specific association for several body fat-related clinical parameters to HSLi6 A5, and to HSLi6 A5 homozygosity, comparing A5 with the other alleles in group. The subjects were 323 obese patients (85 males, 238 females) without other metabolic complication, and 301 nonobese healthy individuals (134 males, 167 females). They were analyzed for various body fat-related clinical parameters, and HSLi6 genotype. RESULTS: Homozygosity for HSLi6 A5 was a risk factor for obesity, BMI > or = 30 kg m-2 (Odds ratio = 1.75, 95% CI 1.58-1.93) and body fat mass > 39.6% (Odds ratio = 1.89, 95% CI 1.60-2.23) in women. This genotype was also associated with increased diastolic blood pressure and triglyceride level among nonobese women, and with increased body fat mass and waist/hip ratio among nonobese men. CONCLUSION: HSLi6 A5 homozygosity is a risk factor for body fat accumulation. PMID- 12534455 TI - P53 codon 72 polymorphism and susceptibility to development of human papilloma virus-associated cervical cancer in Indian women. AB - BACKGROUND: Infection with human papilloma virus (HPV) is an important etiological factor in the development of cervical cancer and it has been proposed that individuals homozygous for Arg/Arg at codon-72 of p53 are seven times more susceptible to HPV-mediated cancer. In this study, we have analyzed the genetic predisposition of the India population to HPV infection and cervical carcinogenesis. METHODS: We investigated 71 cases of squamous cell carcinoma of the cervix, 14 cases of low-grade squamous intraepithelial lesions, 25 cases of high-grade squamous intraepithelial lesions and 29 noncancer controls for presence of HPV16/18 infections by L-1-specific PCR assay and Southern hybridization, and its association with polymorphism at p53 codon 72. RESULTS: We observed that 69.1% (76/110) of the cervical cancer patients were HPV positive, among which the presence of HPV16, 18 and 16/18 coinfection was 40.9%, 8.2% and 13.6%, respectively. The allele frequencies of the three p53 genotypes Arg/Arg, Arg/Pro and Pro/Pro in the HPV-positive tumour samples were 0.34, 0.57 and 0.09 in comparison with frequencies of 0.18, 0.44 and 0.38 for HPV-negative tumours. Hence, there is a significant difference in the allelic frequency of p53 Arg/Arg in high-risk HPV-infected cervical carcinoma cases (0.34) and HPV-negative carcinomas (0.18). CONCLUSION: Our results indicate a striking over representation of homozygous arginine at codon 72 of p53 in HPV-associated cervical carcinogenesis. We conclude that women with Arg/Arg homozygous allele are more prone to infection by HPV16/18, which leads to cervical carcinomas having poor prognosis. PMID- 12534456 TI - Clinical and genetic features underlying the response of patients with bronchial asthma to treatment with a leukotriene receptor antagonist. AB - BACKGROUND: Treatment with antileukotriene drugs results in clinical improvement in many, though not all, patients with asthma. It can be hypothesized that the subpopulation of asthmatic patients, characterized by aspirin intolerance and cysteinyl-leukotriene overproduction, might profit most from antileukotriene treatment. MATERIALS AND METHODS: We compared the clinical response to montelukast in two well-matched groups of patients with mild asthma: 26 aspirin intolerant asthmatics (AIAs) and 33 aspirin-tolerant asthmatics (ATAs). We also searched for possible predictors of the clinical response among the parameters reflecting the expression and production of cysteinyl-leukotrienes (cys-LTs). This was an 8-week, single-blind, placebo-controlled trial. RESULTS: Following a 3-week montelukast 10 mg day-1 treatment compared with placebo, there was a statistically significant reduction in the mean daytime and nocturnal asthma symptoms and beta 2-agonist use, as well as a significant improvement in the morning and evening peak expiratory flows and quality of life. Both groups showed a similar significant improvement in the parameters studied. Clinical response did not correlate with the baseline urinary LTE4 excretion level. Improvement of asthma was observed mostly in patients with a low baseline and non-IL-5 inducible expression of LTC4 synthase (LTC4S) mRNA in eosinophils. There was a trend toward a better response in carriers of LTC4S allele C, but no relationship to the CC10 genetic polymorphism. CONCLUSIONS: No difference in the clinical response to the montelukast treatment was observed between the AIAs and the ATAs. PMID- 12534457 TI - Airway surface liquid recovered by lavage with perfluorocarbon liquid in cats. AB - BACKGROUND: Airway surface liquid (ASL) is difficult to sample. Lavage with an immiscible perfluorocarbon (PFC) liquid to recover ASL was evaluated in cats. MATERIALS AND METHODS: Six wild-type cats underwent bronchoscopic lavage with a PFC (perfluorohexane), with the bronchoscope wedged in the feline equivalent of the right lower lobe. Two cats (control animals) were lavaged with a saline vehicle only. Four procedures were performed on each animal at 2-3-week intervals. Ionic composition of ASL was determined by flame photometry. RESULTS: Cats lavaged with PFC showed significantly more acute respiratory distress than those lavaged with saline (respiratory rate following procedure: PFC, 47 +/- 5 min-1 vs. saline, 27 +/- 2 min-1, P < 0.05; O2 saturation: PFC 80 +/- 1% vs. saline, 91 +/- 1%, P < 0.01). The PFC group also had clinical evidence of chronic respiratory compromise (mean respiratory rate before next anaesthetic; PFC, 37 +/ 2 min-1 vs. saline, 20 +/- 3 min-1, P < 0.01). The PFC-lavaged lungs demonstrated persistent radiographic changes and histological evidence of small airways obstruction with distal alveolar damage. Six PFC lavages yielded ASL samples (> 100 microL) which were sufficient for analysis. Mean (+/- SEM) ionic concentrations in these samples were Na+ 157.4 +/- 14.5 mmol L-1, Cl- 150.5 +/- 16.8 mmol L-1 and K+ 10.1 +/- 1.7 mmol L-1. CONCLUSIONS: Perfluorocarbon lavage can be used to collect unmodified ASL from the distal lung. However, repeated lavage with perfluorohexane was associated with significant pathological changes in this study. PMID- 12534460 TI - Pseudomonas putida: a cosmopolitan opportunist par excellence. PMID- 12534461 TI - Travels of a Pseudomonas, from Japan around the world. PMID- 12534462 TI - Diversity of siderophore-mediated iron uptake systems in fluorescent pseudomonads: not only pyoverdines. AB - Fluorescent pseudomonads are gamma-proteobacteria known for their capacity to colonize various ecological niches. This adaptability is reflected by their sophisticated and diverse iron uptake systems. The majority of fluorescent pseudomonads produce complex peptidic siderophores called pyoverdines or pseudobactins, which are very efficient iron scavengers. A tremendous variety of pyoverdines has been observed, each species producing a different pyoverdine. This variety can be used as an interesting tool to study the diversity and taxonomy of fluorescent pseudomonads. Other siderophores, including newly described ones, are also produced by pseudomonads, sometimes endowed with interesting properties in addition to iron scavenging, such as formation of complexes with other metals or antimicrobial activity. Factors other than iron limitation, and different regulatory proteins also seem to influence the production of siderophores in pseudomonads and are reviewed here as well. Another peculiarity of pseudomonads is their ability to use a large number of heterologous siderophores via different TonB-dependent receptors. A first genomic analysis of receptors in four different fluorescent pseudomonads suggests that their siderophore ligand repertoire is likely to overlap, and that not all receptors recognize siderophores as ligands. PMID- 12534463 TI - Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440. AB - Pseudomonas putida is a metabolically versatile saprophytic soil bacterium that has been certified as a biosafety host for the cloning of foreign genes. The bacterium also has considerable potential for biotechnological applications. Sequence analysis of the 6.18 Mb genome of strain KT2440 reveals diverse transport and metabolic systems. Although there is a high level of genome conservation with the pathogenic Pseudomonad Pseudomonas aeruginosa (85% of the predicted coding regions are shared), key virulence factors including exotoxin A and type III secretion systems are absent. Analysis of the genome gives insight into the non-pathogenic nature of P. putida and points to potential new applications in agriculture, biocatalysis, bioremediation and bioplastic production. PMID- 12534464 TI - Global features of the Pseudomonas putida KT2440 genome sequence. AB - The compositional bias of the G+C, di- and tetranucleotide contents in the 6 181 862 bp Pseudomonas putida KT2440 genome was analysed in sliding windows of 4000 bp in steps of 1000 bp. The genome has a low GC skew (mean 0.066) between the leading and lagging strand. The values of GC contents (mean 61.6%) and of dinucleotide relative abundance exhibit skewed Gaussian distributions. The variance of tetranucleotide frequencies, which increases linearly with increasing GC content, shows two overlapping Gaussian distributions of genome sections with low (minor fraction) or high variance (major fraction). Eighty per cent of the chromosome shares similar GC contents and oligonucleotide bias, but 105 islands of 4000 bp or more show atypical GC contents and/or oligonucleotide signature. Almost all islands provide added value to the metabolic proficiency of P. putida as a saprophytic omnivore. Major features are the uptake and degradation of organic chemicals, ion transport and the synthesis and secretion of secondary metabolites. Other islands endow P. putida with determinants of resistance and defenceor with constituents and appendages of the cell wall. A total of 29 islands carry the signature of mobile elements such as phage, transposons, insertion sequence (IS) elements and group II introns, indicating recent acquisition by horizontal gene transfer. The largest gene carries the most unusual sequence that encodes a multirepeat threonine-rich surface adhesion protein. Among the housekeeping genes, only genes of the translational apparatus were located in segments with an atypical signature, suggesting that the synthesis of ribosomal proteins is uncoupled from the rapidly changing translational demands of the cell by the separate utilization of tRNA pools. PMID- 12534465 TI - The genome structure of Pseudomonas putida: high-resolution mapping and microarray analysis. AB - As part of a collaborative project aimed at sequencing and functionally analysing the entire genome of Pseudomonas putida strain KT2440, a physical clone map was produced as an initial resource. To this end, a high-coverage cosmid library was arrayed and ordered by clone hybridizations. Restriction fragments generated by rare-cutting enzymes and plasmids containing the rrn operon and 23S rDNA of Pseudomonas aeruginosa were used as probes and, parts of the cosmids were end sequenced. This provided the information necessary for merging and comparing the macro-restriction map, cosmid clone order and sequence information, thereby assuring co-linearity of the eventual sequence assembly with the actual genome. A tiling path of clones was selected, from the shotgun clones used for sequencing, for the production of DNA microarrays that represent the entire genome including its non-coding portions. PMID- 12534466 TI - Genomic analysis of the aromatic catabolic pathways from Pseudomonas putida KT2440. AB - Analysis of the catabolic potential of Pseudomonas putida KT2440 against a wide range of natural aromatic compounds and sequence comparisons with the entire genome of this microorganism predicted the existence of at least four main pathways for the catabolism of central aromatic intermediates, that is, the protocatechuate (pca genes) and catechol (cat genes) branches of the beta ketoadipate pathway, the homogentisate pathway (hmg/fah/mai genes) and the phenylacetate pathway (pha genes). Two additional gene clusters that might be involved in the catabolism of N-heterocyclic aromatic compounds (nic cluster) and in a central meta-cleavage pathway (pcm genes) were also identified. Furthermore, the genes encoding the peripheral pathways for the catabolism of p hydroxybenzoate (pob), benzoate (ben), quinate (qui), phenylpropenoid compounds (fcs, ech, vdh, cal, van, acd and acs), phenylalanine and tyrosine (phh, hpd) and n-phenylalkanoic acids (fad) were mapped in the chromosome of P. putida KT2440. Although a repetitive extragenic palindromic (REP) element is usually associated with the gene clusters, a supraoperonic clustering of catabolic genes that channel different aromatic compounds into a common central pathway (catabolic island) was not observed in P. putida KT2440. The global view on the mineralization of aromatic compounds by P. putida KT2440 will facilitate the rational manipulation of this strain for improving biodegradation/biotransformation processes, and reveals this bacterium as a useful model system for studying biochemical, genetic, evolutionary and ecological aspects of the catabolism of aromatic compounds. PMID- 12534467 TI - Detection of multiple extracytoplasmic function (ECF) sigma factors in the genome of Pseudomonas putida KT2440 and their counterparts in Pseudomonas aeruginosa PA01. AB - Pseudomonas putida KT2440 is highly successful in colonizing a variety habitats, including aquatic and edaphic niches. In accordance with this ability and with the need to adapt to changing environmental conditions, P. putida has developed sophisticated mechanisms of transcriptional regulation. We analysed, at the genome level, the repertoire of sigma factors in P. putida KT2440 and identified 24 sigma factors, 19 of which corresponded to the subfamily of extracytoplasmic function (ECF) sigma factors. We detected 13 ECF sigma factors that showed similarity to the Escherichia coli FecI sigma factor, which is involved in iron acquisition. In 11 cases, a fecR-like gene was found adjacent to the fecI-like gene and, in 10 cases, a gene encoding an iron receptor lies in the vicinity of the fecI/fecR cluster. This may explain the ability of P. putida KT2440 to grow under low iron availability conditions. Five fecI/fecR/iron receptor gene clusters from P. putida were also identified in the human pathogen Pseudomonas aeruginosa. PMID- 12534468 TI - Complete sequence of the IncP-9 TOL plasmid pWW0 from Pseudomonas putida. AB - The TOL plasmid pWW0 (117 kb) is the best studied catabolic plasmid and the archetype of the IncP-9 plasmid incompatibility group from Pseudomonas. It carries the degradative (xyl) genes for toluenes and xylenes within catabolic transposons Tn4651 and Tn4653. Analysis of the complete pWW0 nucleotide sequence revealed 148 putative open reading frames. Of these, 77 showed similarity to published sequences in the available databases predicting functions for: plasmid replication, stable maintenance and transfer; phenotypic determinants; gene regulation and expression; and transposition. All identifiable transposition functions lay within the boundaries of the 70 kb transposon Tn4653, leaving a 46 kb sector containing all the IncP-9 core functions. The replicon and stable inheritance region was very similar to the mini-replicon from IncP-9 antibiotic resistance plasmid pM3, with their Rep proteins forming a novel group of initiation proteins. pWW0 transfer functions exist as two blocks encoding putative DNA processing and mating pair formation genes, with organizational and sequence similarity to IncW plasmids. In addition to the known Tn4651 and IS1246 elements, two additional transposable elements were identified as well as several putative transposition functions, which are probably genetic remnants from previous transposition events. Genes likely to be responsible for known resistance to ultraviolet light and free radicals were identified. Other putative phenotypic functions identified included resistance to mercury and other metal ions, as well as to quaternary ammonium compounds. The complexity and size of pWW0 is largely the result of the mosaic organization of the transposable elements that it carries, rather than the backbone functions of IncP-9 plasmids. PMID- 12534469 TI - Analysis of the Pseudomonas aeruginosa oprD gene from clinical and environmental isolates. AB - Genomes are constantly evolving. Our report highlights the wide mutational diversity of clinical as well as environmental isolates, compared with the laboratory strain(s), through the systematic genetic analysis of a chromosomal porin gene (oprD) in relation to a specific antibiotic resistance. Mutational inactivation of the oprD gene is associated with carbapenem resistance in Pseudomonas aeruginosa. The sequence of the oprD gene of 55 Pseudomonas aeruginosa natural isolates obtained from across the world--from sources as diverse as patients and rhizospheres--was analysed. A microscale mosaic structure for this gene--resulting from multiple intra- and possibly interspecies recombinational events--is reported. An array of independent and seemingly fast occurring defective oprD mutations were found, none of which had been described before. A burn wound isolate demonstrated unusually high overall sequence variability typical of mutator strains. We also present evidence for the existence of OprD homologues in other fluorescent pseudomonads. PMID- 12534470 TI - Pseudomonas aeruginosa O-antigen chain length is determined before ligation to lipid A core. AB - Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen that infects immunocompromised patients and trauma victims and causes fatal lung infections in people with cystic fibrosis. This microorganism produces a number of virulence factors, one of which is lipopolysaccharide (LPS), which has been shown to mediate many biological effects including resistance to serum killing and phagocytosis. These biological activities have been correlated to the length of the O-polysaccharide and its distribution on the outer membrane. Wzz is responsible for regulation of the size distribution of the O-antigen. Wzz has been found to participate solely in the Wzy-dependent pathway for LPS biosynthesis, which produces heteropolymeric O-polysaccharide such as the B-band LPS of P. aeruginosa. Our laboratory has previously reported characterization of a Wzz protein encoded in the B-band O-antigen biosynthesis cluster of PAO1. The availability of the genome sequence of P. aeruginosa PAO1 has made it possible to identify a second functional Wzz protein (PA0938, Wzz2). Gene replacement was used to generate an unmarked wzz2delta knock-out and a wzz2delta/wzz1::Gm double knock-out. As expected, the wzz2delta strain produced LPS with modal length imparted by Wzz1, and the wzz2delta/wzz1::Gm strain produced LPS O-antigen with a non-modal (random) length. Both wzz1 and wzz2 from P. aeruginosa PAO1 were cloned and expressed with an N-terminal His6 tag. His6-Wzz1 and His6-Wzz2 were purified to near homogeneity by immobilized metal affinity chromatography (IMAC). These preparations were used to develop specific polyclonal antibodies against each of the proteins. In vivo protein cross-linking followed by Western immunoblotting indicated that Wzz1 forms dimers whereas Wzz2 forms octamers. By generation of a wzz2delta/rmlC double mutant and analysis of the LPS, we have made the novel observation that polymerization of modal chain length-distributed O-antigen occurred before ligation to the lipid A core. We have shown an association between the Wzz proteins and O-antigen polymer chains using immunoprecipitation with anti-O5 O-antigen monoclonal antibody MF15-4. Both Wzz1 and Wzz2 could be co precipitated with O5 polymer. PMID- 12534471 TI - Pseudomonas aeruginosa displays an epidemic population structure. AB - Bacteria can have population structures ranging from the fully sexual to the highly clonal. Despite numerous studies, the population structure of Pseudomonas aeruginosa is still somewhat contentious. We used a polyphasic approach in order to shed new light on this issue. A data set consisting of three outer membrane (lipo)protein gene sequences (oprI, oprL and oprD), a DNA-based fingerprint (amplified fragment length polymorphism), serotype and pyoverdine type of 73 P. aeruginosa clinical and environmental isolates, collected across the world, was analysed using biological data analysis software. We observed a clear mosaicism in the results, non-congruence between results of different typing methods and a microscale mosaic structure in the oprD gene. Hence, in this network, we also observed some clonal complexes characterized by an almost identical data set. The most recent clones exhibited serotypes O1, 6, 11 and 12. No obvious correlation was observed between these dominant clones and habitat or, with the exception of some recent clones, geographical origin. Our results are consistent with, and even clarify, some seemingly contradictory results in earlier epidemiological studies. Therefore, we suggest an epidemic population structure for P. aeruginosa, comparable with that of Neisseria meningitidis, a superficially clonal structure with frequent recombinations, in which occasionally highly successful epidemic clones arise. PMID- 12534472 TI - Pedigree and taxonomic credentials of Pseudomonas putida strain KT2440. PMID- 12534475 TI - Water: Lifesaver or killer? PMID- 12534476 TI - The prehospital airway: more questions or more answers? PMID- 12534477 TI - Decompression sickness. PMID- 12534478 TI - Accidental arterial gas embolism. PMID- 12534479 TI - What you need to know about diving medicine but won't find in a textbook. AB - The old adage that 'if a patient in your emergency department (ED) is wearing a wetsuit, fins and a mask, then he/she probably has a diving related illness' is one that should be remembered. This is an obvious statement that should not need stating; however, simple clues can be missed or disregarded. This article will address issues that may confront emergency physicians and for which there are few resources to find the answers. It aims to explain the reasons behind some of the advice given during consultation with a hyperbaric physician. The second aim is to bring emergency physicians up to date with new diving practices and how these may impact upon traditional diving injuries. To achieve these aims, this article is a compilation of answers to frequently asked or pertinent questions related to diving medicine. PMID- 12534480 TI - Drowning and near-drowning. AB - Water immersion is a frequent cause of accidental death and hospital admission. This article outlines the pathogenesis and principles of treatment. Drowning is defined as death by asphyxia due to submersion in a liquid medium. Near-drowning is defined as immediate survival after asphyxia due to submersion. PMID- 12534481 TI - Survey of major chronic iIlnesses and hospital admissions via the emergency department in a randomized older population in Randwick, Australia. AB - OBJECTIVE: To find out if patients with chronic illnesses living in the community are at risk of unplanned hospital admissions through emergency departments; what types of chronic illnesses may be putative risk factors; and if an increase in the number of chronic illnesses may be associated with increased risk. METHODS: The survey included the completion of a standardized questionnaire for medical illnesses in a random sample of older people dwelling in the community and analysis of admission records to our hospital. The principal diagnoses for admissions were recorded. The risk factors for admissions were analysed. RESULTS: Five hundred and twenty-six (239 men and 287 women) people aged 55 years and over were interviewed. Musculoskeletal disorders, hypertension, gastrointestinal disorders and ischaemic heart disease were the most frequently reported of the chronic illnesses surveyed. A total number of 70 people from the survey group with a total of 115 admissions through emergency departments were recorded. Using logistic regression model, hypertension, ischaemic heart disease and age were found to be risk factors for emergency admissions amongst this group of community dwelling residents. The ratios were 2.03 (95% confidence interval (CI): 1.2 3.44), 2.02 (95% CI: 1.16-3.49) and 1.05 (95% CI: 1.02-1.09), respectively. Furthermore, multiple (three or more) chronic illnesses were found to be a strong predictor of hospital admission via emergency department (chi-square = 16.647, DF = 1, P-value < 0.001). CONCLUSION: We conclude that there was significant association between multiple chronic diseases and emergency admissions for older people. Of these, hypertension and ischaemic heart disease were found to be significant predictors. Age per se was found to be of borderline significance. PMID- 12534482 TI - Validation of a tool to safely triage selected patients with chest pain to unmonitored beds. AB - OBJECTIVE: To externally validate a chest pain protocol that triages low risk patients with chest pain to an unmonitored bed. METHODS: Retrospective study of all patients admitted from the emergency department of a tertiary referral public teaching hospital with an admission diagnosis of 'unstable angina' or suspected ischemic chest pain. Data was collected on adverse outcomes and analysed on the basis of intention-to-treat according to the chest pain protocol. RESULTS: There were no life-threatening arrhythmias, cardiac arrests or deaths within the first 72 h of admission in the group assigned to an unmonitored bed by the chest pain protocol ([0/244]; 0.0%: 95% confidence interval 0.0-1.5%). Four patients had an uncomplicated myocardial infarction, two patients had recurrent ischemic chest pain and one patient developed acute pulmonary oedema ([7/244]; 2.9%: 95% confidence interval 1.2-5.8%). CONCLUSION: This retrospective study externally validated the chest pain protocol. Care in a monitored bed would not have altered outcomes for patients triaged to an unmonitored bed by the chest pain protocol. Compared to current guidelines, application of the chest pain protocol could increase the availability of monitored beds. PMID- 12534483 TI - Adverse events and complications among patients admitted to hospital in the home directly from the emergency department. AB - INTRODUCTION: The nature and incidence of adverse events and complications among patients admitted from the emergency department to hospital in the home has not been investigated. This study aimed to investigate this problem and make recommendations for prevention strategies. METHODS: This was an explicit retrospective chart review of patients admitted from the emergency department directly to hospital in the home between 22 February 1995 and 1 September 2000. A data extraction document was designed specifically for the study and used to extract data relating to patient demographics, diagnosis, past medical history and outcome. The outcomes of interest include adverse events, complications and death. An adverse effect is defined as an unintended injury or complication that results in disability, death or prolonged hospital stay and is caused by health care management. These adverse events may occur prior to or during the index admission and may be noted during or after the index admission. A complication is defined as an undesirable outcome that occurs during the management but not causing disability, death or prolonged hospital stay. RESULTS: Three hundred and fifty-seven patients were enrolled (51.3% male; median age 52 years, range 16-96 years). Fifty-five adverse events were identified: 49 adverse events (89%) were due to management prior to hospital in the home admission and six adverse events (10.9%) were directly attributable to hospital in the home management. This represents a rate of 1.7 adverse events per 100 hospital in the home admissions directly attributable to hospital in the home management. One hundred and eighteen complications were identified. Most complications were easily managed. Thirty-one patients had unplanned re-admissions and two patients died within 28 days of hospital in the home admission. CONCLUSION: Most patients admitted to hospital in the home from the emergency department were managed successfully. Few adverse events arose from hospital in the home treatment. Complications were common but minor in nature. Strategies for the prevention of phlebitis and constipation are recommended. PMID- 12534484 TI - The use of rapid sequence intubation by ambulance paramedics for patients with severe head injury. AB - OBJECTIVE: To determine the effects of rapid sequence intubation in patients with severe head injury performed by paramedics on a helicopter emergency medical service. METHODS: The patient care records for patients with severe head injury who underwent rapid sequence intubation between November 1999 and February 2002 (inclusive) were examined. Data were extracted on the demographics of the patients, as well as the physiological changes before and after rapid sequence intubation. RESULTS: There were 122 patients with severe head injury evaluated at the scene during the study period. Rapid sequence intubation was attempted in 110 patients and was successful in 107 (97%). Intubation was associated with improvements in systolic blood pressure, oxygen saturation and end-tidal carbon dioxide levels, compared with baseline levels. CONCLUSION: Rapid sequence intubation in patients with severe head injury may be safely undertaken by helicopter-based ambulance paramedics and is associated with improvements in oxygenation, ventilation and blood pressure. Further studies of this skill undertaken by road-based paramedics are warranted. PMID- 12534485 TI - Blunt carotid artery injury. AB - Blunt carotid artery injury is an uncommon injury with a potentially devastating outcome. Although treatment is often unhelpful for an established neurologic deficit, there is frequently a delay between the episode of trauma and the onset of neurology. This delay provides a window of opportunity where early detection and intervention may significantly improve outcome and prevent lifelong morbidity in a generally young trauma patient population. This paper will review the literature regarding incidence, outcome, treatment options and optimal methods of diagnosis for this lesion. The following synopsis of a case seen at Royal Perth Hospital Emergency Department illustrates several of the classical features of blunt carotid injury, including delayed presentation and pattern of associated injuries. The patient, a 19-year-old female, presented following a single vehicle motor crash. On arrival she had a Glasgow Coma Scale (GCS) of 14 with no focal neurologic deficit. Multiple injuries were identified, including a large scalp laceration, a compound fracture of the mandible, pneumothorax, sternal fracture, subcapsular haematoma of the liver, and a knee laceration. Initial computed tomography (CT) scan of the head was normal. Twenty hours after the motor vehicle accident the patient's conscious state deteriorated to GCS 9. Repeat CT head demonstrated changes consistent with a left middle cerebral artery infarct. Cerebral angiography revealed traumatic dissection of the left internal carotid artery with complete occlusion of the lumen just distal to its origin. After 13 days in the intensive care unit and 2.5 months in the rehabilitation unit, she was discharged with a persistent severe receptive and expressive aphasia, as well as a dense right hemiparesis. She is now living at home with community supports. PMID- 12534486 TI - Amiodarone: an emergency medicine perspective. AB - Amiodarone is a highly efficacious antiarrhythmic agent for many cardiac arrhythmias, ranging from atrial fibrillation to malignant ventricular rhythm disturbances. Significant interest has developed in recent years with the publication of randomized controlled trials supporting the efficacy of amiodarone over placebo and lignocaine for improving survival to hospital in patients with shock-resistant ventricular fibrillation. Amiodarone has complex pharmacological and pharmacokinetic properties. It has significant long-term adverse effects, but short-term administration of intravenous amiodarone is generally well tolerated. This article will explore issues related to the clinical use of amiodarone from an emergency medicine perspective. PMID- 12534487 TI - Benchmarking in emergency health systems. AB - This paper discusses the role of benchmarking as a component of quality management. It describes the historical background of benchmarking, its competitive origin and the requirement in today's health environment for a more collaborative approach. The classical 'functional and generic' types of benchmarking are discussed with a suggestion to adopt a different terminology that describes the purpose and practicalities of benchmarking. Benchmarking is not without risks. The consequence of inappropriate focus and the need for a balanced overview of process is explored. The competition that is intrinsic to benchmarking is questioned and the negative impact it may have on improvement strategies in poorly performing organizations is recognized. The difficulty in achieving cross-organizational validity in benchmarking is emphasized, as is the need to scrutinize benchmarking measures. The cost effectiveness of benchmarking projects is questioned and the concept of 'best value, best practice' in an environment of fixed resources is examined. PMID- 12534488 TI - Failure of intramuscular antivenom in Red-back spider envenoming. AB - Four cases of Red-back spider envenoming are reported in which there was minimal response to intramuscular antivenom. Intravenous antivenom was then administered in each case with almost complete resolution of symptoms. All cases were followed up to confirm the effect of treatment. This failure of intramuscular Red-back antivenom raises the question of its efficacy. There has been no controlled trial to prove that intramuscular Red-back antivenom is effective and animal work with other antivenoms has demonstrated the intramuscular formulation to have delayed and incomplete effects. Controlled studies should be undertaken to establish the effectiveness of intravenous and intramuscular Red-back antivenom. PMID- 12534489 TI - The toxicity of antidepressant poisoning: is it changing? A comparative study of cyclic and newer serotonin-specific antidepressants. AB - AIM: To compare the clinical features of cyclic antidepressant and newer, non cyclic, serotonin-specific antidepressant poisoning. METHODS: Comparitive, descriptive study of all antidepressant overdose patients admitted to a hospital toxicology service from February 1997 to April 2001. Patient data were entered prospectively into a dedicated toxicology database for subsequent analysis. RESULTS: There were 256 admissions for antidepressant poisoning (17.5% of all poisoning admissions). Cyclic antidepressant poisoning comprised 43% of antidepressant admissions. Statistically significant differences between the two groups included: cyclic antidepressant group had longer median length of stay (23.1 vs 15.9 h, P = 0.0008), greater need for endotracheal intubation (31%vs 4%, OR = 11.5, P < 0.0001) and higher incidence of seizures (7.2%vs 0.7%, OR = 10.4, P = 0.01), faster median pulse rate, longer QRS-interval on admission, and longer intensive care unit stays. However, non-cyclic, serotonin-specific antidepressant poisonings involved larger doses of antidepressants and were more likely to ingest other medications along with these. Serotonin syndrome was only seen in non-cyclic, serotonin-specific poisoning (10.3%, OR = 26.6, P = 0.0002). Patients with serotonin syndrome had a longer median hospital stay (46 vs 16 h, P < 0.0002) compared to other non-cyclic, serotonin-specific patients. There were no deaths during the study period. CONCLUSIONS: Cyclic antidepressants still comprise a significant proportion of antidepressant poisoning and result in more significant morbidity than non-cyclic, serotonin-specific poisoning. Clinicians should also be aware that non-cyclic, serotonin-specific poisoning may result in the development of serotonin syndrome. This was the most significant toxic effect noted following non-cyclic, serotonin-specific poisoning in this study. PMID- 12534490 TI - Intracerebral haemorrhage presenting as olfactory hallucinations. AB - Isolated olfactory hallucinations are a rare event and have been associated with a number of aetiologies including seizures, migraines and psychiatric illnesses. This case report describes a 58-year-old woman with an unusual haemorrhagic lesion as the cause of her olfactory symptoms. A review of the literature concerning the causes and management recommendations for olfactory hallucinations is presented. PMID- 12534498 TI - Quality management in emergency medicine. PMID- 12534504 TI - Efficacy, safety, and cost of office-based surgery: a multidisciplinary perspective. AB - An increasing number of media reports on patient safety risks arising from office based surgery procedures, as well as growing concerns about patient safety issues in general, have brought office-based surgery as well as its practitioners into focus and placed this very cost-effective medical practice in the eye of the media and regulators. Concerted efforts are now being made to understand the causes and true incidence of patient safety risk associated with office-based surgery and to find ways to minimize this risk. PMID- 12534505 TI - Comparison of botulinum toxin types A and B: a bilateral and double-blind randomized evaluation in the treatment of canthal rhytides. AB - BACKGROUND: The obligate bacterium Clostridium botulinum produces exotoxins (A, B, C-1, C-2, D, E, F, G) that are serologically and antigenically distinct. All serotypes have similar neurotoxic properties resulting in flaccid muscle paralysis. Types A and B are commercially available and are used widely for the reduction of dynamic facial rhytides. Although extensive information is known about type A, type B has recently become available; however, there is a limited clinical familiarity. Some of the remaining unknown distinctions between the two subtypes are the extent of toxin diffusion from the site of injection, the onset of action, the dose equivalency, and the duration of effect. OBJECTIVE: The purpose of this preliminary double-blind study was to compare the duration of muscle paralysis and rhytid reduction of botulinum toxin types A and B. Additional relevant information was obtained, all of which can be useful in toxin selection. METHOD: Ten women, ages 28 through 60, voluntarily consented to undergo a double-blind trial and were randomly assigned to have botulinum toxin type A injected into one set of lateral canthal rhytides and toxin type B into the contralateral periocular region. Based on dose-ranging investigations performed in patients with cervical dystonia, participants received treatment at the lowest reported effective ratio of 1:50 (1 U of toxin type A to 50 U of toxin type B). Three injections of 5 U of type A (total 15 U) and three injections of 250 U of undiluted type B (total 750 U) were injected into the lateral fibers of the orbicularis oculi muscle. RESULTS: Patients were evaluated at 7, 30, 60, and 90 days. Findings were compared by the treating physician, patient self assessment, and photographic images. CONCLUSION: All patients noted rapid and satisfactory reduction in the rhytides in both periocular areas. However, upon unblinding of the solutions at the same volumes with a 1:50 ratio, type B toxin was found to be associated with slightly more discomfort upon injection, quicker onset of action, a sensation of "tightness" of the treated area, and a briefer duration of muscle paralysis. PMID- 12534506 TI - Scar quality and physiologic barrier function restoration after moist and moist exposed dressings of partial-thickness wounds. AB - BACKGROUND: There is growing evidence of improved healing of full- and partial thickness cutaneous wounds in wet and moist environments. Retention of biologic fluids over the wound prevents desiccation of denuded dermis or deeper tissues and allows faster and unimpeded migration of keratinocytes over the wound surface. It allows also the naturally occurring cytokines and growth factors to exert their beneficial effect on wound contracture and re-epithelialization. Despite all of these documented benefits, applying the moist healing principles to large surface areas, in particular to large burns, is hindered by the major technical handicap of creating and maintaining a sealed moist environment over these areas. METHODS: From January to September 2001, healing of partial thickness skin graft donor sites was studied in a prospective comparative study of two types of moist dressings, Tegaderm (3M Health Care, St. Paul, MN), a semipermeable membrane occlusive dressing, and moist exposed burn ointment (MEBO) (Julphar; Gulf Pharmaceutical Industries, United Arab of Emirates), an ointment that can provide a moist environment without the need of an overlying occlusive dressing. Healing was assessed both clinically and with serial measurements of transepidermal water loss (TEWL) and moisture. Following healing, scar quality was evaluated by two members of the team separately using a visual analog scale. Results were statistically analyzed. RESULTS: Faster healing was observed clinically with MEBO application. Physiologic healing as determined by TEWL measurements occurred at an extremely significant earlier stage for MEBO, and this was associated with better scar quality, demonstrating a positive relationship between function and cosmetic appearance. Moreover, the ointment is definitely easier to apply than the occlusive self-adhesive membrane, which requires some degree of dexterity and expertise. CONCLUSION: MEBO application is an effective and valid alternative to conventional occlusive dressings. Moreover, the observed improved anatomic and physiologic healing indicates that MEBO may have a positive effect on healing more that the mere fact of passive moisture retention. PMID- 12534507 TI - Indications for lymph node dissection in the treatment of extramammary Paget's disease. AB - BACKGROUND: In extramammary Paget's disease, compared with patients with carcinoma in situ (CIS), patients with malignancies underlying dermis and subcutaneous tissues or lymph node metastasis appear to have a much worse prognosis. OBJECTIVE: The purpose was to examine our extramammary Paget's diseases patients in terms of pathological tumor invasion, lymph node metastasis, and prognosis and to refer to indications of lymph node dissection. METHODS: The 34 patients were divided into four groups by pathological tumor invasion: CIS, microinvasion to the papillary dermis (MIPD), invasion to the reticular dermis (IRD), and invasion to subcutaneous tissues (IST). Their age, sex ratio, follow up periods, history of other cancers, lymph node metastasis, and prognosis were compared among the four groups and referred to indications of lymph node dissection. RESULTS: In the groups of CIS and MIPD, all patients appeared to have no lymph node metastasis and death caused by this disease. In the groups of IRD and IST, 8 of 12 patients showed pathological lymph node metastasis, and all of them died due to metastasis of the tumor. In the cumulative survival rate of the patients according to the level of invasion, 5-year survival rate was 100% in CIS and MIPD groups, 33.3% in the IRD group, and 0% in the IST group. CONCLUSION: In the treatment of extramammary Paget's disease, indications of lymph node dissection are an important issue as well as wide local excision. From this prospective study, we have developed an algorithm for indications of lymph node dissection. PMID- 12534508 TI - Laser scar revision: comparison study of 585-nm pulsed dye laser with and without intralesional corticosteroids. AB - BACKGROUND: Hypertrophic scars affect 1.5% to 4.5% of the general population and remain notoriously difficult to eradicate because of the high recurrence rates and the incidence of side effects associated with treatment. Pulsed dye laser (PDL) treatment and intralesional corticosteroids have individually been reported to be effective in reducing hypertrophic scar bulk and symptoms. OBJECTIVE: To determine whether combination PDL and intralesional corticosteroid treatment produces better hypertrophic scar improvement than PDL treatment alone. METHODS: Bilateral hypertrophic inframammary scars in 22 females were randomly assigned to receive treatment with 585-nm PDL alone or in combination with intralesional corticosteroid. Clinical evaluations and scar pliability scores were determined before each of the two treatment sessions and 6 weeks after the final treatment. Histologic evaluation of skin biopsies obtained before and after treatment was performed in four patients. RESULTS: All scars showed clinical improvement with increased pliability and decreased symptoms (pruritus) after each of the two treatments. Clinical improvement scores were not significantly better with the concomitant use of corticosteroids. Side effects were limited to mild purpura and transient hyperpigmentation. Decreased sclerosis was seen in scars after PDL treatment (with or without concomitant corticosteroids). CONCLUSIONS: Treatment of hypertrophic inframammary scars with 585-nm PDL irradiation alone effected substantial clinical and histologic improvement. The adjunctive use of intralesional corticosteroids did not significantly enhance clinical outcome except in those scars that were most symptomatic. PMID- 12534509 TI - The use of a new diode laser for hair removal. AB - BACKGROUND: Diode and long-pulse 1064 Nd:YAG lasers, because of their long wavelengths and extended pulse durations, have gained popularity over the last several years for photoepilation in individuals with dark skin phenotypes. OBJECTIVE: To analyze the clinical efficacy and histologic features of a new 810 nm diode laser in the treatment of unwanted hair in individuals with Fitzpatrick skin types II-IV. METHODS: Twenty-four female subjects (mean age, 33; range, 17 53) were treated three times at monthly intervals with the MeDioStara diode laser (Zeiss-Meditech, Jenna, Germany): wavelength 810 nm, spot size 12 mm, pulse duration 50 msec; fluence 25-35 J. Patients had skin types II-IV and light to dark brown hair. Hair counts were carried out using handheld magnifiers at baseline and at months 1, 2, 3, and 6. Six individuals had biopsies taken after treatment and at 3 and 6 months. RESULTS: A mean hair removal efficiency of 74% and 79% was noted at 3 and 6 months, respectively, and was best in those individuals with skin type III. Hair counts after treatment and at last follow-up were significantly lower when compared with the baseline counts (P<0.0001). Histologic analysis revealed a range from early catagen induction to complete follicular destruction. CONCLUSIONS: This study supports the clinical photoepilatory efficiency of a new 810-nm diode laser in producing 70% clinical hair removal efficiency at 6 months. Histopathologic evaluation suggests that this decrease in hair counts is probably secondary to heat trauma to the follicular epithelium, resulting in complete follicular destruction and stopping of anagen phase. PMID- 12534510 TI - Algorithm for using a long-pulsed Nd:YAG laser in the treatment of deep cutaneous vascular lesions. AB - BACKGROUND: Conventional therapies for deep cutaneous vascular anomalies have demonstrated poor efficacy and many side effects. New laser systems offer greater potential to treat these difficult lesions, but the lack of specific treatment guidelines has restricted consistent success. OBJECTIVE: To establish a rational, user-friendly algorithm that incorporates basic components of deep vascular lesions to define the correct laser settings required for safe, effective, and reproducible treatment. METHODS: Within 18 months, 162 deep vascular lesions of various types and anatomic sites were evaluated for vessel size, depth, color, and pressure. An algorithm incorporating these characteristics was employed to determine laser parameter settings. Using a high-peak power, long-pulse 1064-nm Nd:YAG laser system, the vascular lesions were then treated. RESULTS: Within 6 months of follow-up, 80% of treated areas demonstrated a 50% or greater resolution after a single treatment session, with complete clearance shown in 19%. Only minimal and transient side effects were observed. Of note, 74% of areas on the extremities and 83% within the oral cavity showed a 50% or greater resolution after one treatment. CONCLUSION: Previously challenging deep cutaneous vascular anomalies may be safely reduced or cleared with the use of an appropriate laser system and this algorithm-directed technique. This represents a significant breakthrough in the management of vascular lesions. PMID- 12534511 TI - Evaluation of the role of exogenous estrogen in postoperative progress after laser skin resurfacing. AB - BACKGROUND: Recent studies indicate that exogenous estrogens may promote wound healing. Many laser skin resurfacing (LSR) patients use hormone replacement therapy (HRT) or oral contraceptive pills (OCPs). OBJECTIVE: To evaluate the effect of exogenous estrogen on LSR postoperative healing. METHODS: This is a retrospective case control study of 44 female patients who underwent combination CO2/Er:YAG full-face LSR. Sixteen postmenopausal patients using oral HRT during procedure and follow-up were compared with 16 controls. Six premenopausal patients on estrogen-containing OCPs during the procedure and follow-up were compared with six controls. Case and control groups were matched by age, skin type, and treatment technique. Premenopausal and postmenopausal groups were compared. The variables evaluated included erythema, swelling, crusting, purpura, pain, pruritus, reepithelialization, complications, and patient assessment of outcome. RESULTS: For premenopausal and postmenopausal women, there were no significant differences in outcome measures of postoperative morbidity between groups receiving and not receiving estrogen. The incidence of complications, time until re-epithelialization, and patient assessment of outcome were similar between groups. CONCLUSION: Our findings suggest that exogenous estrogen in the form of HRT or OCPs does not clinically alter postoperative progress in combination CO2/Er:YAG LSR patients. Premenopausal women heal comparably to postmenopausal women after LSR. PMID- 12534512 TI - Long-term psychological impact and perceived efficacy of pulsed-dye laser therapy for patients with port-wine stains. AB - BACKGROUND: The rationale for treatment of patients with port-wine stain (PWS) birthmarks has focused on the belief that eventual soft-tissue hypertrophy and negative psychological effects might be diminished or avoided. Pulsed-dye laser (PDL) therapy has become widely accepted as the treatment of choice for PWS, and a variety of studies have been conducted that evaluate its short-term efficacy. Long-term data regarding outcome and patient satisfaction are lacking. OBJECTIVE: Our objective was to evaluate the long-term efficacy of pulsed-dye laser therapy from the patient's perspective. METHODS: A survey was mailed to all patients or parents of minor patients whose last known laser treatment was performed between January 1, 1989, and January 1, 1996. Patients were asked to quantify changes in their PWS as well as their psychological well-being. A formal qualitative analysis was performed on their written comments as well. RESULTS: A total of 164 surveys were mailed to the most recent address available by the patient record. Fifty-two surveys were returned as undeliverable, and 55 evaluable surveys were returned, yielding a 49% overall return rate for patients who could be located. The mean age of respondents was 28.8 years, 96% of whom were white and 62% female. The mean number of years since last treatment was 7.04. The vast majority of patients noted little or no change in texture, height, or dimension of their PWS, whereas 62% noted color improvement. A majority or patients (60%) worried less about their appearance after treatment, whereas a similar number (61%) believed their ability to make friends or meet others was unaffected by treatment. Only 19% thought others looked at or treated them differently because of their PWS. Overall, 48% of patients indicated satisfaction with treatment, 24% dissatisfaction, and 28% neutral. On a 10-point scale indicating their likelihood of recommending treatment to someone similarly affected, the mean score was 7.42. Men were significantly and consistently less satisfied with treatment than women, despite rating the degree of color improvement similarly to women. Qualitative analysis of patient comments helped validate quantitative data and revealed gender differences in satisfaction, as well as correlations with adverse events from treatment, desire for additional treatment, and a trend toward more positive comments with the passage of time. CONCLUSION: The pulsed-dye laser improves the color of PWS over long periods of time in a majority of patients. Patients tended to worry less about their appearance after treatment, although most believed treatment did not substantially affect their relationship with others or others' view of them. Most patients were satisfied or neutral with regard to satisfaction with therapy and would recommend treatment to others. A minority of patients was dissatisfied with treatment, and men were more likely to be dissatisfied. Additional long-term and prospective studies will be helpful in assessing the physical and psychosocial impact of PDL for PWS. PMID- 12534513 TI - Millisecond 1064-nm neodymium:YAG laser treatment of facial telangiectases. AB - BACKGROUND: Millisecond pulse duration 1064-nm Neodymium:YAG (Nd:YAG) lasers have been shown to be effective in the treatment of some lower leg telangiectases. OBJECTIVE: To evaluate the efficacy and complication rate of a millisecond pulse duration 1064-nm Nd:YAG laser in the treatment of facial telangiectases. METHODS: Fifteen subjects were evaluated. RESULTS: Moderate to significant improvement was seen in 73% patients at day 30 and in 80% of patients at 3 months. These results were seen in the treatment of both blue/red and red facial telangiectases. CONCLUSION: The millisecond pulse duration 1064-nm Nd:YAG laser is effective for treatment of facial telangiectases. PMID- 12534514 TI - Combined ultrapulse CO2 laser and Q-switched alexandrite laser compared with Q switched alexandrite laser alone for refractory melasma: split-face design. AB - BACKGROUND: : Melasma is common and can cause major psychological impact. To date, the mainstay of treatment, including various hypopigmenting agents and chemical peels, is ineffective and can cause adverse effects. Laser is a new approach and is yet to be explored for its efficacy and safety. OBJECTIVE: : To compare combined Ultrapulse CO2 laser and Q-switched alexandrite laser (QSAL) with QSAL alone in the treatment of refractory melasma. METHODS: : Six Thai females were treated with combined Ultrapulse CO2 laser and QSAL on one side of the face and QSAL alone on the other side. The outcome was evaluated periodically for up to 6 months using the modified Melasma Area and Severity Index score and the modified Melasma Area and Melanin Index score. RESULTS: : The side with combination treatment had a statistically significant reduction of both scores. On the QSAL side, the score reduction was not significant. Two cases developed severe postinflammatory hyperpigmentation and were effectively treated with bleaching agents. Transient hypopigmentation and contact dermatitis were observed with the combination treatment side. CONCLUSIONS: : Combined Ultrapulse CO2 laser and QSAL showed a better result than QSAL alone but was associated with more frequent adverse effects. Long-term follow-up and a larger number of cases are required to determine its efficacy and safety for refractory melasma. PMID- 12534515 TI - 585-nm pulsed dye laser in the treatment of surgical scars starting on the suture removal day. AB - BACKGROUND: The optimal time frame to improve the quality and cosmetic appearance of scars by laser therapy has not been clearly elucidated by prior controlled clinical trials. OBJECTIVE: To determine the efficacy of the 585-nm pulsed dye laser (PDL) in the treatment of surgical scars starting on the day of suture removal. METHODS: Eleven patients (skin types I-IV) with 12 postoperative linear scars that were greater than 2 cm were treated three times on monthly intervals with the 585-nm PDL (450 micro s, 10-mm spot size, 3.5 J/cm2 with 10% overlap) on one scar half, whereas the other half received no treatment. Scars were later evaluated by a blinded examiner using the Vancouver Scar Scale (VSS) for pigmentation, vascularity, pliability, and height. Scars were then blindly examined for cosmetic appearance using a visual analog scale. RESULTS: One month after the last treatment, final scar analysis by the blinded examiner revealed a significant difference between treated and untreated sites, with the treated halves scoring better in all scar parameters in the VSS and in cosmetic appearance. The treated halves demonstrated an overall average improvement in the VSS between the first treatment score and the final score of 54% versus 10% in the controls (P=0.0002). The cosmetic appearance score (0=worst; 10=best) at final assessment was significantly better for the treated scars, scoring 7.3 versus the averaged control score of 5.2 (P=0.016). CONCLUSION: The 585-nm PDL is effective and safe in improving the quality and cosmetic appearance of surgical scars in skin types I-IV starting on the day of suture removal. PMID- 12534516 TI - Transconjunctival laser blepharoplasty of lower eyelids: Asian experience with 1,340 cases. AB - BACKGROUND: Lower eyelid bulging is a result of the aging process, but it can sometimes be severe in young people. This condition can cause considerable aesthetic problems to many patients. OBJECTIVE: To determine the effectiveness of transconjunctival laser blepharoplasty in treating Asian skin. METHODS: Since 1995, our clinic has performed 1,340 procedures to remove the fat bulging of lower eyelids using a carbon dioxide laser. RESULTS: The most common complaints were edema and transient discomfort and red eye; 82% of the patients were satisfied (1,099 patients rated a score of 3 or 4). The incidence of complications was very low. CONCLUSION: Transconjunctival laser blepharoplasty of lower eyelids is a safe, low-risk procedure with excellent cosmetic results. It makes no visible scar and reduces downtime. This is also good for patients with previous surgical scars on their lower eyelid. PMID- 12534517 TI - Single-pass carbon dioxide versus multiple-pass Er:YAG laser skin resurfacing: a comparison of postoperative wound healing and side-effect rates. AB - BACKGROUND: Ablative laser skin resurfacing with carbon dioxide (CO2) and erbium:yttrium-aluminum-garnet (Er:YAG) lasers has been popularized in recent years and their side effects individually reported. No prior study, however, has directly compared the relative healing times and complications rates between the two different systems. OBJECTIVE: To evaluate and compare postoperative wound healing and short- and long-term side effects of single-pass CO2 and multiple pass, long-pulsed Er:YAG laser skin resurfacing for the treatment of facial photodamage and atrophic scars. METHODS: A retrospective chart review and analysis of sequential clinical photographs were performed in 100 consecutive patients who underwent laser skin resurfacing with single-pass CO2 (Ultrapulse 5000; Coherent, Palo Alto, CA, N=50) or multiple-pass, long-pulsed Er:YAG laser resurfacing (Contour; Sciton, Palo Alto, CA, N=50). All laser procedures were performed by a single operator for the amelioration of facial rhytides or atrophic scars. The rate of re-epithelialization, duration of erythema, and presence of complications were tabulated. RESULTS: The average time to re epithelialization was 5.5 days with single-pass CO2 and 5.1 days with long-pulsed Er:YAG laser resurfacing. Postoperative erythema was observed in all patients, lasting an average of 4.5 weeks after single-pass CO2 laser treatment and 3.6 weeks after long-pulsed Er:YAG laser treatment. Hyperpigmentation was seen in 46% of the patients treated with single-pass CO2 and 42% of the patients treated with the long-pulsed Er:YAG laser (average duration of 12.7 and 11.4 weeks, respectively). No incidences of hypopigmentation or scarring were observed. CONCLUSION: Skin resurfacing with single-pass CO2 or multiple-pass long-pulsed Er:YAG laser techniques yielded comparable postoperative healing times and complication profiles. PMID- 12534518 TI - Treatment of trichostasis spinulosa in skin phototypes III, IV, and V with an 800 nm pulsed diode laser. AB - BACKGROUND: : Trichostasis spinulosa (TS) is a common follicular disorder that results from the retention of multiple vellus hairs within pilosebaceous follicles. A variety of treatment modalities have been used with variable but largely transient success. OBJECTIVES: : To determine whether a pulsed diode laser irradiation would provide a therapeutic response to TS for a prolonged period. METHODS: : Thirteen subjects with untreated TS and skin phototypes III, IV, and V were treated with a 800-nm pulsed diode laser at fluences ranging from 24 to 40 J/cm2 (mean, 36 J/cm2) and a 12- to 20-ms (mean 18 ms) pulse width. Two treatments were delivered at 4-week intervals. Evaluation of improvement was performed at 4 and 20 weeks after the last treatment by a blinded assessment of clinical photographs. RESULTS: : Complete clearing of the lesions was achieved for a period of 8 to 12 weeks. A decrease in dark-plug appearance of greater than 50% was noted in half of the subjects 20 weeks after the second treatment. No pigmentary changes and scarring occurred in any subjects. CONCLUSION: : Pulsed diode laser proved to be a safe and long-term effective treatment for TS in dark skinned individuals. PMID- 12534519 TI - Repair of full-thickness nasal alar defect. PMID- 12534520 TI - Technique of periorbital lipoaugmentation. AB - With aging comes atrophy of the periorbital subcutaneous fat. This leads to abnormal relationships between the orbital and neighboring cosmetic units, with a loss of the anterior projection of the upper third of the face, relative skin excess, and inferior displacement of tissues. Multistage, blunt infiltration with autologous fat is an excellent way to correct these incipient senescent changes. PMID- 12534521 TI - Multicentric malignant melanoma in a giant melanocytic congenital nevus 20 years after dermabrasion in adulthood. AB - BACKGROUND: Dermabrasion is one approach to the treatment of treating giant melanocytic congenital nevi. Treatment is recommended to reduce the risk of spontaneous malignant transformation of giant nevi into malignant melanomas that usually occur in childhood. OBJECTIVE: To describe the development of a multicentric malignant melanoma in a giant melanocytic congenital nevus after dermabrasion. METHODS: We report about a 46-year-old male patient who developed a multicentric malignant melanoma in a giant melanocytic congenital nevus. The nevus was located on his left shoulder extending to his neck and chest. Previously, dermabrasion of the nevus was performed twice at the ages of 26 and 28. RESULTS: To our knowledge, this is the first report of malignant transformation of a giant nevus into a multicentric malignant melanoma diagnosed 20 years after the procedure of dermabrasion. CONCLUSION: We conclude that a close follow-up of such patients is mandatory. PMID- 12534522 TI - Bullous malignant melanoma: an unusual differential diagnosis of a hemorrhagic friction blister. AB - BACKGROUND: A 66-year-old woman presented to our outpatient clinic with a 3 x 2.5 cm tense, hemorrhagic-appearing bulla on her forefoot. Histopathology and immunohistochemistry confirmed a transtumoral-transepidermal blister formation within an advanced acrolentiginous malignant melanoma (MM). OBJECTIVE: To study bullous malignant melanoma. METHODS: Blistering in MMs represents a rare but clinically important pitfall in clinical differential diagnosis. The blisters are typically due to the disruption of the cohesion between neoplastic cells and keratinocytes, but physical friction may also contribute. RESULTS: Hemorrhagic blistering is, in many cases, a relatively insignificant finding in which frictional forces are imposed. CONCLUSION: The case reported here underscores that in rare cases MMs, particularly if acrally located, can be complicated by hemorrhagic blistering. Because of the life-threatening consequences, one should be aware of this rare differential diagnosis. PMID- 12534523 TI - Sebaceous carcinoma arising from nevus sebaceus: a case report. AB - BACKGROUND: Sebaceous carcinoma (SC) is a rare, aggressive, malignant tumor that is derived from adnexal epithelium of sebaceous glands that are distributed mostly in the skin of the head, neck, and hair-bearing regions of the body. OBJECTIVE: To discuss the clinical and histologic features and the prognosis of the extraocular SC. METHOD: We report a case of SCs of the scalp arising from a previous lesion clinically compatible with a nevus sebaceous. The lesion was surgically excised completely. RESULTS: The postoperative course was free of complications, and the patient was clinically free of disease at her recent follow-up 2 years after treatment. CONCLUSIONS: The absence of metastasis or cutaneous recurrence in our patient, notwithstanding the size of the lesion and its late removal, almost 25 years after the first signs of its growth, confirms a much improved prognostic behavior for extraocular SCs. PMID- 12534524 TI - Squamous cell carcinoma of the anal margin with pruritus ani of long duration. AB - BACKGROUND: Anal margin is an unusual location for squamous cell carcinoma (SCC). On rare occasions, anal margin carcinoma is the cause of pruritus ani. OBJECTIVE: To describe a case of SCC of the anal margin with pruritus ani of long duration. RESULTS: A 52-year-old man had been aware of perianal itching for over 10 years. Examination of the perianal area revealed a reddish, eroded, hard nodule that was 2.0 x 1.5 x 0.3 cm in size located in the 3 o'clock position. The histopathologic diagnosis was well-differentiated SCC. The nodule was totally excised with a 1-cm margin. No recurrence or metastases were observed for 7 months. CONCLUSION: When pruritus ani does not respond to conservative therapy and when symptoms have existed for a long time, we should suspect the presence of malignancy. PMID- 12534525 TI - Ocular complication of PhotoDerm VL therapy for facial port-wine stain. AB - BACKGROUND: A case of focal damage to the iris with distortion of the pupil secondary to PhotoDerm therapy in a 2-year-old boy is reported. OBJECTIVE: To study ocular complication of photoDerm VL therapy for facial port-wine stain. METHODS. : Observatory case report. RESULTS: PhotoDerm VL therapy may damage ocular tissues. CONCLUSION: Appropriate protection during the procedure is essential. PMID- 12534526 TI - Microprocessor-controlled local anesthesia versus the conventional syringe technique in hair transplantation. PMID- 12534527 TI - Submental scar revision via cervicomental liposculpturing and submental scar resection. PMID- 12534528 TI - Skin protection programmes. AB - Educational programmes, including evidence-based recommendations on skin protection (skin protection programmes), have been proposed for the prevention of irritant contact dermatitis in occupations with a high frequency of this disease. Recent intervention studies directed at wet-work employees showed a positive influence on wet-work behaviour and on clinical skin symptoms. The structure and implementation of skin protection programmes are discussed. PMID- 12534529 TI - A review of the scientific basis for uncertainty factors for use in quantitative risk assessment for the induction of allergic contact dermatitis. AB - Safety evaluations for chemicals which possess the ability to cause sensitization by skin contact have traditionally been done using an ad hoc comparative risk assessment technique. Recently, several papers have been published supporting the use of an alternative, and potentially better, quantitative risk assessment approach. While they represent a relatively new approach to risk assessment for sensitizers, quantitative methods have been used for decades to support risk assessments for systemic toxicity. Historically, these methods have involved the extrapolation of toxicity data - generally from studies in laboratory animals at relatively high doses to human exposures at lower doses. For toxicity endpoints with a threshold, this process has traditionally involved the use of uncertainty factors. For example, uncertainty factors are commonly used to extrapolate from laboratory animals to humans, and from 'average' humans to sensitive subpopulations. In the absence of data to support a different value, a default factor of 10 is widely accepted for each of these areas. Recent papers have advocated the use of a similar approach to characterize the risk of the induction of skin sensitization by allergens of varying potency and potential for skin contact. As with other forms of toxicity, a quantitative assessment of risk for allergic skin reactions can be approached by identifying a NOAEL (no observed adverse effect level) and applying appropriate uncertainty factors. Three major areas of data extrapolation have been identified: inter-individual susceptibility, the influence of vehicle or product matrix, and exposure considerations. This paper provides an overview of each of these areas with an evaluation of the available scientific database to support an uncertainty factor in the range of 1-10 for each area. PMID- 12534530 TI - Variability in responsiveness to irritants: thoughts on possible underlying mechanisms. AB - Patch testing with chemical irritants almost always produces a striking variability in the intensity of reaction between individuals, even amongst normal, healthy subjects. Whilst there have been many attempts to define factors which predispose to heightened or, conversely, to diminished reactivity, the underlying cellular mechanisms responsible for the variability remain poorly understood. In this review, a number of possible explanations are proposed, with a particular emphasis on those which relate to the influence of pre-existing disease or to the genetic regulation of certain immunological and inflammatory processes. PMID- 12534531 TI - Ethnicity as a possible endogenous factor in irritant contact dermatitis: comparing the irritant response among Caucasians, blacks, and Asians. AB - Irritant contact dermatitis (ICD) is a common dermatological diagnosis in which endogenous and exogenous factors have been implicated. This review explores ethnicity as a possible endogenous factor. While there is a clinical consensus that blacks are less reactive and Asians are more reactive than Caucasians, the data supporting this hypothesis rarely reaches statistical significance. The studies reviewed do not provide sufficient evidence to determine that race is a predisposing factor to the degree of irritation in ICD. We conclude that race could be a factor in ICD, which has practical consequences regarding topical product testing requirements, an ever-expanding global market, occupational risk assessment, and the clinical thinking about ICD. Pitfalls in defining differences are discussed. PMID- 12534532 TI - Further important sensitizers in patients sensitive to fragrances. AB - In order to find sensitizers additional to the current fragrance mix (FM) a series of fragrance materials (series II) was evaluated in 6 dermatological centres in Europe. 11 of the test materials were essential oils, the remaining 7 being either mixtures of isomers or simple chemicals of frequent usage in the perfume industry. 1606 patients were consecutively tested with series II and 8% FM. Each patient was classified regarding a history of adverse reactions to scented products: certain, probable, questionable, none. Reactions to FM occurred most frequently in 11.4% of the subjects. The 6 materials with the highest reactivity after the FM were ylang-ylang oil (YY) I (2.6%), YY II (2.5%), lemongrass oil (1.6%), narcissus absolute (1.3%), jasmine absolute (1.2%) and sandalwood oil (0.9%). 48 (3.0%) of the patients reacted only to materials of series II and not to FM. 6.0% of 1606 patients gave a history of adverse reactions to fragrances which was classified as certain. This group reacted to FM only in 22.9%, to series II and FM in 15.6% and to series II only in 5.2%. 63.5% of the patients reacting to both FM and 1 of the materials of series II had some type of positive fragrance history, which was higher in comparison to those with isolated reactions to FM (46.2% of 121) or to series II, respectively, (45.8% of 48). However, this difference was not statistically significant. In conclusion, the materials of series II identified a further subset of patients with a fragrance problem, which would have been missed by the current FM as the single screening tool for patch testing. PMID- 12534533 TI - Gas chromatography: an investigative tool in multiple allergies to essential oils. AB - Essential or fragrant oils are volatile odourous mixtures of organic chemical compounds that are widely used in aromatherapy and in the perfume industry. Because of their frequent use, allergy to essential oils is being increasingly recognized. We report 2 cases of multiple allergies to essential oils in professional aromatherapists. Gas chromatography/mass spectrometry was used to analyse the oils in order to identify a common allergen responsible for the contact dermatitis. In both the cases, alpha- and beta-pinene were found to be the most common constituent in the oils and thus appeared to be key allergens. alpha-pinene was confirmed as an allergen on repeat patch testing with pure alpha pinene in both cases. 12 controls tested were negative for the same. Gas chromatography-mass spectrometry was found to be an extremely useful tool that could be utilized in investigating multiple allergies to essential oils. PMID- 12534534 TI - Sensitizing capacity and cross-reaction pattern of the isomers of diglycidyl ether of bisphenol F in the guinea pig. AB - In this study, the sensitizing capacities of the 3 main low-molecular-weight isomers of epoxy resins based on diglycidyl ether of biphenol F (DGEBF) were investigated in a guinea pig maximization test (GPMT). The isomers are p,p' DGEBF, o,p'-DGEBF, and o,o'-DGEBF. The sensitization capacities were compared to diglycidyl ether of bisphenol A (DGEBA), which is the main constituent in the most common epoxy resin. The cross-reaction pattern between all 4 compounds was also investigated. It was found that all 3 DGEBF isomers were potent sensitizers. It was also found that p,p'-DGEBF, o,p'-DGEBF, and DGEBA cross-reacted with one another, and that animals induced with o,o'-DGEBF reacted significantly only to o,p'-DGEBF, but not to the other 2 DGEBF isomers or DGEBA. PMID- 12534535 TI - 55 cases of allergic reactions to hair dye: a descriptive, consumer complaint based study. AB - Severe facial and scalp dermatitis following the use of permanent hair dyes has been reported in several cases. Para-phenylenediamine (PPD) is known as a potent contact allergen, and PPD is allowed in hair dye at a concentration of 6%. Hair dye reactions are usually diagnosed by the patients themselves, and adverse reactions to hair dye may not necessarily be recorded by the health care system, unless the reactions are especially severe. Based on this assumption, we suspected that hair dye dermatitis was occurring more frequently than reported in the literature. Consumer complaint-based data were obtained by advertising for persons with adverse reactions to hair dye. Among those responding to the advertisement, 55 cases of severe, acute allergic contact dermatitis were identified. The main symptoms were severe oedema of the face, scalp and ears, and clinically this was often mistaken for angio-oedema. The 55 cases comprised a total of 75 visits to the health service and 5 admissions to hospital. 18 persons had sick leave, which supports the impression of very severe dermatitis reactions. 60% were treated with antihistamine, while 52% were treated with corticosteroids. 29% of the cases were patch tested and all were found positive to PPD. Our data presented here clearly show that PPD and its derivatives in hair dye at the present concentrations presents a significant health risk for the population. Furthermore, the severe acute allergic skin reactions are often misdiagnosed in the health care system. The frequency of allergic contact dermatitis resulting from hair dye is likely to be underestimated. New methods to survey the frequency of adverse reactions should be considered. PMID- 12534536 TI - Occupational allergic contact dermatitis caused by isocyanates. AB - Between 1978 and 2001, 22 patients were diagnosed with occupation-related allergic contact dermatitis from isocyanates and/or polyurethanes in our clinic. 13 had a positive reaction to the isocyanates, of whom 10 also reacted to diaminodiphenylmethane (MDA), which is used in the production or processing of isocyanates and polyurethanes; 9 reacted only to MDA. The object of the present study was to identify the trades and industries responsible for the development of contact allergy to these resins. Such patients must be patch tested with the isocyanates contacted at work, and account must be taken of positive reactions to MDA as a marker for isocyanate sensitivity. PMID- 12534537 TI - Allergic contact dermatitis from melphalan and chlorambucil: cross-sensitivity or cosensitization? PMID- 12534538 TI - Urticaria after topical immunotherapy with diphenylcyclopropenone. PMID- 12534539 TI - Immediate and delayed hypersensitivity to ceftriaxone, and anaphylaxis due to intradermal testing with other beta-lactam antibiotics, in a previously amoxicillin-sensitized patient. PMID- 12534540 TI - Contact allergy to Tween 80 in an inhalation suspension. PMID- 12534541 TI - Analysis of immunoglobulin E-mediated responses in central Africa. PMID- 12534542 TI - The immunobiology of allergenic peptidases. PMID- 12534543 TI - Microarrayed recombinant allergens for diagnosis of allergy. AB - We suggest that the coapplication of recombinant allergens and microarray technology can lead to the development of new forms of multi-allergen tests which allow the determining and monitoring of complex sensitization profiles of allergic patients in single assays. The allergen extracts which have so far been used for diagnosis only allowed the determining of whether an allergic patient is sensitized against a particular allergen source, but the disease-eliciting allergens could not be identified. Through the application of recombinant DNA technology a rapidly growing panel of recombinant allergen molecules has become available which meanwhile comprises the epitope spectrum of most of the important allergen sources. We demonstrate that microarray technology can be used to establish multi-allergen tests consisting of microarrayed recombinant allergen molecules. Microarrayed recombinant allergens can be used to determine and monitor the profile of disease-eliciting allergens using single tests that require minute amounts of serum from allergic patients. The wealth of diagnostic information gained through microarray-based allergy testing will likely improve diagnosis, prevention and treatment of allergy. PMID- 12534544 TI - Increased airway inflammatory cells in endurance athletes: what do they mean? AB - BACKGROUND: Inflammatory cells are increased in the airways of endurance athletes, but their role in causing exercise-induced respiratory symptoms and bronchoconstriction, or their possible long-term consequences, are uncertain. AIM: To put the results of athlete studies in perspective, by analysing the pathogenesis of airway cell changes and their impact on respiratory function. RESULTS: Athletes of different endurance sports at rest showed increased airway neutrophils. Elite swimmers and skiers also showed large increases in airway eosinophils and lymphocytes, possibly related to chronic, exercise-related exposure to irritants or cold and dry air, respectively. Post-exercise studies reported variable responses of airway cells to exercise, but found no evidence of inflammatory cell activation in the airways, at variance with exercise-induced neutrophil activation in peripheral blood. The increase in airway inflammatory cells in athletes can result from hyperventilation-induced increase in airway osmolarity stimulating bronchial epithelial cells to release chemotactic factors. Hyperosmolarity may also inhibit activation of inflammatory cells by causing shedding of adhesion molecules, possibly explaining why airway inflammation appears 'frustrated' in athletes. Data on exhaled nitric oxide are few and variable, not allowing conclusions about its usefulness as a marker of airway inflammation in athletes, or its role in modulating bronchial responsiveness. CONCLUSIONS: The acute and long-term effects of exercise on airway cells need further study. Airway inflammatory cells are increased but not activated in athletes, both at rest and after exercise, and airway inflammation appears to regress in athletes quitting competitions. Altogether, these findings do not clearly indicate that habitual intense exercise may be detrimental for respiratory health. Rather, airway changes may represent chronic adaptive responses to exercise hyperventilation. An improved understanding of the effects of exercise on the airways will likely have a clinical impact on sports medicine, and on the current approach to exercise-based rehabilitation in respiratory disease. PMID- 12534545 TI - Analysis of the sensitization profile towards allergens in central Africa. AB - BACKGROUND: Almost no information is available regarding the prevalence of IgE mediated allergies and the disease-eliciting allergens in tropical Africa. OBJECTIVE: To study IgE-mediated allergies and the allergen profile in allergic patients from Zimbabwe. METHODS: The frequency of sensitization to common environmental allergen sources was determined by skin prick testing in 650 allergic patients from Zimbabwe. Fifty representative sera were analysed for IgE reactivity to 20 respiratory and 20 food allergen extracts by multiallergen extract testing. The IgE reactivity profiles to recombinant pollen and mite allergens were compared between grass pollen- and mite-sensitized patients from Zimbabwe and central Europe. Sera from grass pollen-allergic patients were also analysed for IgE reactivity to nitrocellulose-blotted natural timothy grass and Bermuda grass pollen allergens. RESULTS: IgE-mediated allergies were found to be common in Zimbabwe. Similar to the situation in central Europe, mites and grass pollens represented the most prevalent allergen sources. However, the IgE reactivity profiles determined with single recombinant pollen and mite allergens revealed interesting differences between the European and African patients, which most likely reflect the local allergen exposure. CONCLUSIONS: The striking differences regarding sensitization to grass pollen and mite allergens between African and European patients revealed by recombinant allergen-based testing emphasize the need for component-resolved allergy testing to optimize allergy prevention and therapy in different populations. PMID- 12534546 TI - Cloning and expression of Blo t 1, a novel allergen from the dust mite Blomia tropicalis, homologous to cysteine proteases. AB - BACKGROUND: House dust mite allergens have been shown to be a very important stimulus in the causation of asthma and triggers for the exacerbation of symptoms. Therefore, characterization of mite-derived allergens at the molecular level is an important step for the development of effective diagnostic and therapeutic approaches, as well as for epidemiological studies. OBJECTIVE: To clone, express and characterize at the molecular level the cysteine protease from Blomia tropicalis (Bt). METHODS: A full-length cDNA encoding Blo t 1 was cloned from a Bt cDNA library using a PCR and RACE-based strategy. The cDNA was PCR amplified, sequenced and subcloned into a prokaryotic expression vector. The allergenicity of the recombinant Blo t 1 was evaluated for IgE reactivity by Western blot. RESULTS: Blo t 1 cDNA encodes a 221 amino acids polypeptide with an estimated molecular weight of 25 kDa. The recombinant protein is 35% identical to other mite cysteine proteases. Recombinant Blo t 1 (rBlo t 1) bound IgE from 62% of Bt skin test-positive serum. Dermatophagoides pteronyssinus (Dp) skin test positive sera did not react with rBlo t 1, indicating the possible presence of unique IgE epitopes on the rBlo t 1 molecule. A three-dimensional image of Blo t 1, constructed based on predicted analysis, showed conserved secondary and tertiary structure with other cysteine proteases. CONCLUSION: We report the cloning, expression and IgE reactivity of Blo t 1, a novel allergen from the domestic mite Blomia tropicalis (Bt), highly homologous to cysteine proteases. This putative cysteine protease, designated Blo t 1, may play a major role as an immunodominant allergen involved in dust mite-specific IgE-mediated hypersensitivity. PMID- 12534547 TI - German cockroach extract increases bronchial epithelial cell interleukin-8 expression. AB - BACKGROUND: Cockroach exposure has been recognized as a common trigger for asthma. While dust mite and Aspergillus fumigatus aeroallergens have been noted to have direct effects on airway epithelium, direct effects of cockroach proteins have not been determined. OBJECTIVE: The purpose of this study was to investigate whether cockroach extract has a direct pro-inflammatory effect on airway epithelium. METHODS: We examined the effect of crude German cockroach (Blattella germanica) extract on IL-8 expression in a human bronchial epithelial cell line (16HBE14o-cells) and primary human bronchial epithelial cells. Transcription from the IL-8 promoter and protein abundance were determined by reporter assay and enzyme-linked immunosorbent assay (ELISA), respectively. Endotoxin levels in the crude cockroach extracts were determined using the Limulus Amebocyte Lysate assay. Protease activity was assessed using Azocoll as a substrate. RESULTS: We found that crude cockroach extract induced a synergistic increase in TNF-alpha induced transcription from the IL-8 promoter. The synergistic effect was observed with as little as 0.3 micro g/mL of crude cockroach extract, while larger concentrations (30 micro g/mL) approximately doubled TNF-alpha-induced IL-8 promoter activity. Similar effects of cockroach extract on IL-8 protein abundance were observed in both 16HBE14o- and primary human bronchial epithelial cells. Addition of endotoxin at concentrations found in the cockroach extract had no effect on TNF-alpha-mediated IL-8 expression. The serine protease inhibitors aprotinin and phenylmethylsulphonyl fluoride abolished cockroach-induced synergy, while the cysteine protease inhibitors E64 and leupeptin had little effect. Measurement of protease activity using Azocoll as a substrate confirmed the presence of protease activity in cockroach extracts. Addition of recombinant Bla g 2, Bla g 3 and Bla g 5 had no effect on TNF-alpha-induced IL-8 promoter activation. Finally, cockroach extract also increased TNF-alpha-induced transcription from the IL-6 promoter. CONCLUSIONS: German cockroach extract contains novel serine protease activity which has a direct pro-inflammatory effect on airway epithelial cells PMID- 12534549 TI - Allergen-specific conventional immunotherapy decreases immunoglobulin E-mediated basophil histamine releasability. AB - BACKGROUND: Allergen-specific immunotherapy has proven to be clinically effective in the treatment of patients with atopic asthma; however, the mechanisms are still unclear. Several noted immunological changes include an increase of the allergen-specific IgG antibody, a reduction in the allergen-specific IgE antibody subsequent to transient increase, an allergen-specific T cell shift in cytokine production from Th2 to Th1, and a decrease in quantity and activity of basophils and mast cells. OBJECTIVE: To analyse the changes of basophil histamine release in response to IgE-mediated and non-IgE-mediated stimuli before and after conventional house-dust mite immunotherapy in children who suffer from atopic asthma. METHODS: Fourteen Dermatophagoides farinae (Df) sensitive asthmatic children with conventional immunotherapy were examined. Basophil histamine releasability was measured 0 months (just before immunotherapy), 4 months and 9 months after immunotherapy. Basophils were stimulated with Df and goat anti-human IgE antibody as IgE-mediated stimuli; and formyl-Met-Leu-Phe (fMLP) and calcium ionophore A23187 as non-IgE-mediated stimuli. Accordingly, the asthma symptom score was used to assess clinical outcome and the skin test reactivity to Df was measured. RESULTS: In contrast to pre-immunotherapy activity, 4 and 9 months after immunotherapy there were significant decreases in histamine release by Df and by anti-IgE antibody. The histamine release by fMLP and by calcium ionophore showed no significant changes after immunotherapy. Histamine release by Df demonstrated significant correlation to that by anti-IgE antibody and by fMLP, yet there was no observable correlation between histamine release by Df and by calcium ionophore. The asthma symptom score decreased significantly 4 and 9 months after immunotherapy and showed significant correlation with histamine release by Df. The skin test reactivity (allergen/histamine ratio) remained constant 4 months after immunotherapy, but decreased significantly 9 months after immunotherapy. CONCLUSION: Basophils have the potential to play an important role in the early clinical improvement of conventional immunotherapy in children with atopic asthma, which may be a result of the decreased IgE-mediated histamine releasability during immunotherapy. PMID- 12534548 TI - Skin test with a timothy grass (Phleum pratense) pollen extract vs. IgE to a timothy extract vs. IgE to rPhl p 1, rPhl p 2, nPhl p 4, rPhl p 5, rPhl p 6, rPhl p 7, rPhl p 11, and rPhl p 12: epidemiological and diagnostic data. AB - BACKGROUND: The diagnostic approach to grass pollen allergy is now possible by detecting specific IgE to its allergenic components. OBJECTIVE: To compare the IgE reactivity to a timothy grass pollen extract with the IgE reactivity to eight allergenic components from the same source (Phl p 1, 2, 4, 5, 6, 7, 11, 12). Both were compared with the skin test reactivity to a timothy grass extract. METHODS: A population survey was carried out by means of the skin test to identify grass allergic subjects, and to characterize them in terms of demographic and allergological parameters. Seven hundred and forty-nine sera were available for IgE detection to a timothy extract, to the recombinant Phl p 1, 2, 5, 6, 7, 11, 12, and to native Phl p 4 and bromelain. Results were stratified by means of demographic and allergy parameters. RESULTS: Ninety-five per cent of the sera had detectable IgE to the timothy extract. Prevalence of IgE reactivity increased from 86.8% to 93.3% as the number of combined reactive molecules rose from 2 to 8. Adjusted prevalences for each allergen were: rPhl p 1 = 83%, rPhl p 2 = 55%, nPhl p 4 = 70%, rPhl p 5 = 50%, rPhl p 6 = 44%, rPhl p 7 = 7%, rPhl p11 = 43%, rPhl p 12 = 15%. Isolated reactivity to rPhl p 1 was 6%, whereas it was negligible for the remaining molecules. IgE reactivity prevalence and mean values differed when patients were stratified on the basis of their associated pollen reactivity and their skin test reactivity grade. No differences were found when age, symptom type and duration were considered. Up to eight-fold higher IgE concentrations were found when the sum of IgE to molecules was compared with IgE to the extract. Testing for the IgE reactivity to the glycan of the native Phl p 4 allergen showed a possible interference with prevalence and value estimation. Higher prevalence values were found in previously immunotherapy-treated patients. CONCLUSIONS: The use of a complete panel of grass allergenic molecules can mimic the current use of allergenic extracts, but new relevant information, such as individual pattern of reactivity, adjusted prevalence, correct specific IgE concentration, can be achieved only by means of discrete allergenic molecules. PMID- 12534550 TI - Hymenoptera ultra-rush venom immunotherapy (210 min): a safety study and risk factors. AB - BACKGROUND: In this study, which summarizes our last 5 years of experience, we evaluated the side-effects of ultra-rush venom immunotherapy and the possibility to define some risk factors for side-effects as age, Hymenoptera venom used for treatment, treatment phase, severity of prior insect sting reaction, concentration of skin test positivity, and level of specific IgE. METHODS: In our protocol on day 1, an initial venom dose of 0.1 microg was followed by 1, 10, 20 microg at 30-min intervals and then 30 and 40 microg at 60-min intervals. Patients who completed this protocol received two booster injections of 50 microg on day 15 and one of 100 microg on day 45. Subsequently, monthly 100 microg boosters were given. RESULTS: Fifty-one children (9.20 +/- 3.41 years) and 207 adults (40.62 +/- 14.00 years) underwent an ultra-rapid venom immunotherapy (ultra-RVIT). Single ultra-RVIT was administered to 195 patients: 69 with honeybee, 123 with yellow jacket and three with wasp venoms. Two venoms were injected into 59 patients: 42 with yellow jacket and wasp, 17 with yellow jacket and honeybee. Four patients received the three venoms. The frequency of immediate systemic reactions (SR) was not significantly different between adults and children: 11.2% vs. 10.8%. SR were experienced more frequently on day 1 (n= 33). They were uncommon on day 15 (n= 2) and on day 45 (n= 1). No late reactions have been observed. Honeybee venom induced significantly more SR (30%) vs. yellow jacket (3.2%) and wasp venom (6.1%). Among the 33 patients who experienced a SR on day 1, 24 had a reaction grade 1 or 2 and nine had a reaction grade 3 or 4. There is a significant risk for patients with a prior reaction grade 3 or 4 to experience a SR during venom immunotherapy (VIT). The strength of positive skin tests and the level of specific IgE were not related to an increased risk of SR (P= NS). CONCLUSION: Treatment with honeybee extract induced more SR than the treatment with yellow jacket and wasp venom. Doses increase phase on day 1 is risk factors for SR of ultra-RVIT, as well as the severity of the prior reaction. Age, degree of positive skin tests, and specific IgE are not risk factors. PMID- 12534551 TI - Effect of various stabilizing agents on Imperata cylindrica grass pollen allergen extract. AB - BACKGROUND: Allergen extracts are unstable, heat labile or susceptible to proteases. Stability of allergen extracts is important for proper diagnosis and therapy of allergic disorders. OBJECTIVE: The present study was undertaken to determine the preservation and stabilization conditions of Imperata cylindrica (Ic) grass pollen extract. METHODS: The Ic extract was kept with 0.1 mepsilon aminocaproic acid (EACA), 0.75 m sucrose, 5% glycerol, 0.03% human serum albumin (HSA) or 0.4% phenol for different time periods. The extracts were stored for 3, 6 and 12 months each at 4 degrees C, 4 degrees C with daily exposure to room temperature (RT) for 1 h, and RT. The quality of extracts was analysed by SDS PAGE, Western blot, ELISA, ELISA inhibition and skin test. RESULTS: Extracts kept with EACA and sucrose retained most of the protein bands followed by glycerol as determined by SDS-PAGE and Western blot during all storage periods and conditions in comparison with standard extracts. The extracts kept with HSA, phenol and without preservative (WP) showed protein degradation below 33 kDa after 3 months storage at all conditions. However, a 67-kDa allergen was stable in these extracts. EACA extract required 75 to 120 ng of protein for 50% inhibition in IgE binding under different conditions, whereas standard extract required 70 ng for the same. ELISA also demonstrated high allergenic reactivity of EACA extract. ID test on allergy patients with EACA extract demonstrated same allergenic potency as that of standard extract. CONCLUSION: EACA is the best preservative/stabilizing agent of Ic pollen extract, followed by sucrose and glycerol. Ic extract kept with phenol, HSA and without preservative showed degradation within 3 months. EACA preserved extract is equally potent as that of standard extract up to 1 year's storage. PMID- 12534552 TI - Immunoglobulin E-binding and skin test reactivity to hydrophobin HCh-1 from Cladosporium herbarum, the first allergenic cell wall component of fungi. AB - BACKGROUND: For many years, fungal spores have been recognized as potential causes of respiratory allergies. All fungal allergens cloned so far represent either secreted or cytoplasmatic proteins, but nothing is known about the involvement of fungal surface proteins in allergic diseases. METHODS: A phage surface displayed cDNA-library from the mould Cladosporium herbarum was constructed and phage displaying IgE-binding proteins were selectively enriched with immobilized serum IgE from C. herbarum-sensitized individuals. Inserts encoding putative allergens were sequenced, subcloned and used to produce recombinant proteins. Allergenicity of the proteins was evaluated by IgE binding in Western blots, enzyme-linked immunosorbent assay (ELISA) and skin prick test in a total of 84 patients sensitized to either C. herbarum or Aspergillus fumigatus and three healthy controls. RESULTS: After four rounds of affinity selection, the cDNA-library was enriched for clones displaying IgE-binding molecules. Sequencing of inserts showed that one clone contained an open reading frame predicting a protein of 105 amino acids and a calculated molecular weight of 10.5 kDa showing the classical signature of members of the hydrophobin family. The recombinant protein, termed HCh-1, was able to bind IgE from six patients sensitized to fungi in vitro. Two of those patients were also included in a skin prick test survey and showed strong type I skin reactions to HCh-1, demonstrating the allergenic nature of C. herbarum hydrophobin and indicating a prevalence of sensitization in the range of 8-9%. In contrast, the hydrophobin HYP1 from Aspergillus fumigatus was not recognized by the sera of the same patients and controls investigated with HCh-1. CONCLUSION: C. herbarum hydrophobin represents the first component of the cell wall of fungi demonstrated to act as a rare but clinically relevant allergen in vitro and in vivo. PMID- 12534553 TI - Interleukin-10 gene promoter region polymorphism is associated with eosinophil count and circulating immunoglobulin E in adult asthma. AB - BACKGROUND: IL-10 has several functional effects relevant to asthma. It can modulate IgE production and induce apoptosis in eosinophils. Polymorphisms of IL 10 gene have been shown to affect IL-10 production. OBJECTIVE: To establish whether IL-10 polymorphisms are associated with asthma and phenotype-related characteristics. METHODS: The frequency of three single base exchange polymorphisms (at positions - 1082, - 819 and - 592) and corresponding haplotypes of the IL-10 gene were analysed in 245 adult asthmatic subjects and 405 controls using PCR and restriction fragment length polymorphism (RFLP). The data were assessed for correlations with the eosinophil count, serum IgE and lung function. RESULTS: The IL-10 haplotype frequencies were similar in asthmatics and controls. Eosinophil count median was 2.0- to 3.2-fold higher among asthmatics with rare ATA/ATA genotype than in asthmatics with other genotypes. No such difference was seen in the control group. When analysed by IL-10 haplotype carrier state and gender, male asthmatics with ATA haplotype had 2.8-fold higher serum IgE than those without ATA. A converse association was found in male controls with ATA haplotype, who had 1.9-fold lower serum IgE than their ATA-negative counterparts. The high IL-10-producing GCC haplotype was associated with impaired lung function in smoking male controls while in asthmatics no clear effect on lung function was found with any of the haplotypes studied. CONCLUSION: These results suggest that the eosinophil counts and serum IgE are differently regulated by IL-10 genotype in asthmatic and in normal subjects. However, IL-10 polymorphism is not related to susceptibility in asthma. PMID- 12534554 TI - Th1/Th2 profile in peripheral blood in atopic cough and atopic asthma. AB - BACKGROUND: Eosinophilic tracheobronchitis with cough hypersensitivity, abbreviated as atopic cough, is an important cause of chronic cough. The reason for the absence of airway hyper-responsiveness is unknown, differing from asthma, a Th2 cytokine-mediated disorder. OBJECTIVE: To compare the type 1 helper T cell (Th1)/Th2 balance in the peripheral blood from subjects with atopic cough and atopic asthma, we assessed the intracellular cytokine production at the single cell level. METHODS: Thirty-six subjects (10 patients with atopic cough, 18 with atopic asthma, and eight control subjects) were included. Intracellular IL-4 and IFN-gamma were detected in CD4+ T cells by flow cytometry. RESULTS: A significantly lower ratio of IFN-gamma-/IL-4-producing CD4+ T cells after phorbol 12-myristate acetate/ionomycin stimulation was found in patients with atopic cough and atopic asthma compared with normal subjects. In comparison between atopic patients, the ratio of IFN-gamma-/IL-4-producing cells was significantly higher in atopic cough than in atopic asthma. However, the proportion of IL-4 producing CD4+ T cells was significantly higher in patients with atopic asthma than in normal control subjects and no significant difference was detected between patients with atopic cough and normal subjects. No significant difference in the proportion of IFN-gamma-producing cells was found between the subjects. Overall, the total IgE levels were positively correlated to the IL-4-producing cells and inversely correlated to the ratio of IFN-gamma-/IL-4-producing cells. CONCLUSION: These results show the lower degree of Th2 cytokine predominance in atopic cough compared with atopic asthma and suggest the relation between the Th1/Th2 balance and atopic status. PMID- 12534555 TI - Humoral and cellular responses to gliadin in wheat-dependent, exercise-induced anaphylaxis. AB - BACKGROUND: Wheat-dependent, exercise-induced anaphylaxis (WDEIA) is a severe allergy where wheat ingestion together with physical exercise induces anaphylaxis. We have previously shown that patients with WDEIA have IgE antibodies against gliadin proteins and identified omega-5 gliadin (Tri a 19) as a major allergen. OBJECTIVE: The aim of this study was to examine gliadin specific IgG subclass, IgA and IgE antibodies, basophil histamine release and cell-mediated responses in WDEIA. METHODS: Sera and peripheral blood mononuclear cells (PBMC) were obtained from patients with WDEIA and from controls without wheat allergy. Serum antibodies to crude gliadin extract (CGE) and purified omega 5 gliadin were measured by ELISA and basophil reactivity by histamine-release test. Gliadin-induced cell-mediated responses were assessed by lymphocyte proliferation assay, and cytokine mRNA expression with real-time quantitative PCR. RESULTS: All patients with WDEIA, but none of the controls, had IgE antibodies to CGE and omega-5 gliadin. Both allergens released high levels of histamine from the basophils of patients with WDEIA. Levels of IgA antibodies to CGE and omega-5 gliadin were significantly elevated in the patients, but the distribution of IgG subclass antibodies showed no statistically significant differences between the two groups. Proliferative responses of PBMC to CGE were increased in patients with WDEIA, and stimulation of PBMC with CGE caused, both in patients and in controls, a clear induction of IL-10 mRNA. Compared with the controls, induction of IL-10 mRNA expression in patients with WDEIA was significantly (P < 0.01) suppressed. CONCLUSION: These results suggest that, in addition to IgE antibodies against omega-5 gliadin, specific IgA antibodies may be involved in the pathogenesis of WDEIA. Decreased expression of IL-10 mRNA in PBMC during gliadin stimulation may facilitate the development of gliadin specific T cell responses. PMID- 12534556 TI - Smoking, atopy and certain furry pets are major determinants of respiratory symptoms in children: the International Study of Asthma and Allergies in Childhood Study (Ireland). AB - BACKGROUND: Environmental, cultural and health care differences may account for variation among countries in the prevalence of asthma and respiratory symptoms in teenagers. OBJECTIVE: To examine the prevalence of respiratory symptoms and the level of diagnosis, and to compare determinants of asthma and severe wheeze in two countries. METHODS: Self-completion questionnaires based on the International Study of Asthma and Allergies in Childhood (ISAAC) protocol were provided to school children in Ireland (Republic and Northern Ireland). In the Republic of Ireland, all children in classes largely aged 13-14 years from 30 post-primary schools were selected by random sampling stratified by school size, composition and Health Board in Spring 1995. In Northern Ireland, all children largely aged 13-14 years of age from 26 post-primary schools were selected by random sampling stratified by school type, composition and Education and Library Board in Spring 1996. RESULTS: Questionnaires were completed by 2,364 children from Northern Ireland and 2,671 from the Republic, about 90% of those eligible to participate. The prevalences of wheeze at various levels of severity, of diagnosed asthma and of treated wheeze were very similar in Northern Ireland and the Republic of Ireland. A significant proportion of those reporting more severe symptomatology (four or more attacks of wheeze in the past 12 months and/or one or more nights disturbed and/or moderate or greater disruption of daily activities and/or speech restriction due to wheeze) had been neither diagnosed nor treated for asthma (20 37%). To investigate the determinants of the more severe symptomatology of asthma or treated wheeze a series of stepwise multiple regression analyses was performed. A history of atopy, cigarette smoking, the possession of a furry pet other than a dog or cat and age were each independently associated with severe wheeze, whilst atopy, a furry pet (as above) and gender were each independently associated with asthma or treated wheeze. CONCLUSIONS: Cigarette smoking is closely associated with the reporting of significant respiratory symptoms together with atopy and exposure to furry pets. Some 20-37% of severe symptoms were neither diagnosed nor treated as asthma. PMID- 12534557 TI - Clinical effects of air filters in homes of asthmatic adults sensitized and exposed to pet allergens. AB - BACKGROUND: Despite medical advice, many pet-allergic asthma sufferers refuse to remove the pet to which they are sensitized from their home. OBJECTIVE: We aimed to assess the clinical effects of air cleaners in the homes of adult asthma patients sensitized and exposed to cats and/or dogs. METHODS: We performed a randomized, parallel-group study of 30 asthmatic adults sensitized to and sharing a home with cats or dogs. The effects of placing air cleaners in the living room and bedroom for 12 months and using high efficiency particulate air filter vacuum cleaners (active group) were compared with using these vacuum cleaners alone (control group). Measures of airway responsiveness, treatment requirement, lung function, peak flow, reservoir and airborne allergen were recorded before, during and after the interventions. A beneficial clinical response was assessed in terms of a 'combined asthma outcome'. This was defined as a two or more doubling dose improvement in bronchial hyper-reactivity to histamine and/or a reduction in treatment requirement of at least one step change on the British Thoracic Society guidelines for asthma treatment. RESULTS: A beneficial clinical response was observed in 10/15 subjects in the active group compared with 3/15 in the control group after 12 months intervention (P = 0.01). No significant differences between the active and control groups were detected for changes in measures of lung function, reservoir pet allergen and airborne pet allergen during the study. CONCLUSION: Whilst the study design has not allowed complete exclusion of a placebo effect, we believe that this pragmatic study of adult asthmatic patients sensitized and exposed to pets resulted in a small, but significant improvement in combined asthma outcome. PMID- 12534558 TI - Immunoadjuvant properties of glove cornstarch powder in latex-induced hypersensitivity. AB - BACKGROUND: Cornstarch powder present in medical gloves plays an important role in latex-induced hypersensitivity as allergen carrier either, by the inhalation route, by skin contact or by direct contact with mucous membranes. OBJECTIVE: Our objective was to test the hypothesis that cornstarch could act as an immunoadjuvant in immediate type-I latex-induced hypersensitivity. METHODS: Guinea-pigs were sensitized by intraperitoneal route with two different antigens (latex proteins and ovalbumin) with or without cornstarch powder. Airway responsiveness after specific bronchial provocation was evaluated and specific IgG and IgG1 levels were determined by enzyme-linked immunosorbent assay (ELISA). Controls were treated with cornstarch powder or saline alone. RESULTS: Animals sensitized with latex proteins (n = 7 in each group) showed significant bronchoconstriction (P < 0.03) and higher anti-latex antibody levels than the controls (P < 0.005). Guinea-pigs sensitized with latex-contaminated cornstarch had higher levels of specific antibodies than those sensitized with latex alone (P < 0.05). Animals sensitized to latex mixed with cornstarch showed higher bronchospasm than those treated with latex alone (P < 0.003). Animals sensitized to ovalbumin mixed with cornstarch also showed higher antibody and bronchoconstriction levels (P < 0.05) than those immunized with ovalbumin alone but antibody titres were significantly lower than those of the animals treated with ovalbumin and Freund's complete adjuvant (P < 0.01; n = 5 in each group). CONCLUSION: Our findings show that cornstarch powder increases antigen-induced bronchoconstriction and antibody production. This role of immunoadjuvant is not antigen-specific. The cornstarch powder used as donning agent in latex gloves is an allergen carrier and it can enhance latex-induced hypersensitivity. PMID- 12534559 TI - Metalloproteinase-9 is increased after toluene diisocyanate exposure in the induced sputum from patients with toluene diisocyanate-induced asthma. AB - BACKGROUND AND OBJECTIVE: Persistent asthma symptoms are associated with airway inflammation and remodeling, which may be mediated through metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP). The aim of this study was to evaluate MMPs and TIMP involvement in toluene diisocyanate (TDI)-induced asthma. MATERIALS AND METHOD: Induced sputum was collected in eight newly diagnosed TDI-induced asthma subjects (group I) before and 7 h after the TDI and placebo challenges and in 12 subjects with TDI-induced occupational asthma diagnosed 5 years previously with persistent asthma symptoms (group II). Sera was collected in group I at diagnosis, and in group II, they were collected at the time of the study. 12 nonasthmatic healthy subjects were enrolled as controls. MMP-9, MMP-2 and TIMP-1 levels in both sputum and serum were measured by enzyme linked immunosorbent assay (ELISA). Gelatinase activity in the sputum was confirmed by zymographic analysis. RESULTS: The serum TIMP-1 level was significantly higher in asthma patients than in healthy controls (P = 0.01), while MMP-9 level was significantly lower in asthmatic patients (P = 0.03). There was no significant difference in MMP-2 level (P = 0.27). MMP-9 level in the sputum was significantly increased after the TDI challenges (P = 0.01). TIMP-1 level in sputum tended to increase after TDI challenges, but no statistical significance was noted (P = 0.09). MMP-9 and MMP-9/TIMP-1 levels in the sputum were significantly higher in group II than in group I (P = 0.04, P = 0.02) with no significant difference in TIMP-1 level. Minimal amount of MMP-2 was found in sputum. Zymography demonstrated that MMP-9 level increased and active form of MMP 9 was generated after the TDI bronchoprovocation test. CONCLUSION: TDI exposure leads to overproduction of MMP-9, which may induce airway inflammation and remodeling, and then contribute to persistent asthmatic symptoms in TDI-induced asthma. PMID- 12534560 TI - Persistence of bronchopulmonary hyper-reactivity and eosinophilic lung inflammation after anti-IL-5 or -IL-13 treatment in allergic BALB/c and IL 4Ralpha knockout mice. AB - BACKGROUND: Antigen-induced bronchopulmonary hyper-reactivity (BHR) is generally associated with eosinophilia. It involves cytokines produced by Th2 lymphocytes, including IL-4, IL-5 and IL-13, which are implicated in IgE production, eosinophil differentiation and attraction, and related events relevant to allergic inflammation, whose mechanisms remain unclear. OBJECTIVE: To investigate the mechanisms by which Th2 cytokines mediate eosinophilia and subsequent BHR using ovalbumin (OVA)-immunized and OVA-challenged IL-4Ralpha-/- and IL-4-/- mice, which fail to transduce and/or to produce IL-4 and IgE as compared with wild type (WT) mice, and specific neutralizing antibodies. METHODS: On days 0 and 7, mice were immunized subcutaneously (s.c.) with OVA. At day 14, anti-IL-5 or anti-IL-13 antibodies were administered intranasally and/or intravenously before allergenic challenge. Different functional and cellular parameters were studied in vivo and cytokine production was followed with a newly described ex vivo procedure using lung explants. RESULTS: IL-4Ralpha-/- and IL-4-/- mice developed BHR and pulmonary eosinophilia, even though eosinophil recruitment to the bronchoalveolar liquid lavage (BALF) was reduced. In vivo, IL-4-/- and IL-4Ralpha /- mice produced, respectively, no or reduced amounts of IL-5 in the BALF/serum as compared with WT mice, whereas no IL-13 in the BALF was detected. By contrast, ex vivo, surviving lung explants from WT and IL-4-/- or IL-4Ralpha-/- mice produced IL-13 and large amounts of IL-5. The neutralization of IL-5 in vivo (BALF and serum) and ex vivo (from lung explant) in IL-4Ralpha-/- and WT mice failed to suppress BHR and lung eosinophilia, and to modify IL-13 production ex vivo. In addition, neutralization of IL-13 in vivo from lung explant also failed to abrogate BHR and lung eosinophilia, whereas IL-5 was unchanged. CONCLUSION: Antigen-induced BHR can develop independently from IL-4, IL-5 or IL-13 and from the IL-4alpha receptor chain, suggesting a possible novel IL-4, IL-5 and IL-13 independent pathway for the development of BHR in allergic BALB/c mice. The failure of IL-5 or IL-13 antibodies to prevent BHR in IL-4Ralpha-/- mice suggests that neither is indispensable for BHR but does not exclude a role for lung tissue eosinophilia. PMID- 12534561 TI - Constitutive secretion of the granule chymase mouse mast cell protease-1 and the chemokine, CCL2, by mucosal mast cell homologues. AB - BACKGROUND: The mucosal mast cell (MMC) granule-specific beta-chymase, mouse mast cell protease-1 (mMCP-1), is released systemically into the bloodstream early in nematode infection before parasite-specific IgE responses develop and TGF-beta1 induces constitutive release of mMCP-1 by homologues of MMC in vitro. Intraepithelial MMC may also express the chemokine CCL2 (monocyte chemotactic protein-1) during nematode infection but the expression of this chemokine by MMC homologues has not been investigated. OBJECTIVE: To investigate the expression and to compare the mechanisms of constitutive release of the chymase, mMCP-1, and the chemokine, CCL2. METHODS: MMC homologues were generated by culturing bone marrow cells in the presence of TGF-beta1, IL-3, IL-9 and stem cell factor (SCF). The intracellular distribution of mMCP-1 and CCL2 was examined by confocal microscopy. The involvement of the Golgi complex and of protein synthesis in the constitutive release of mMCP-1 and CCL2 was investigated using the Golgi disrupting agent brefeldin A and cycloheximide to block protein synthesis. Secreted analytes were quantified by ELISA. RESULTS: mMCP-1 colocalized with Golgi matrix protein 130 but was most abundant in the granules, whereas CCL2 was not found in the granules but appeared to be located uniquely in the Golgi complex. Extracellular release of mMCP-1 was significantly inhibited ( approximately 40%) by cycloheximide and by the Golgi-disrupting agent brefeldin A, indicating both continuous protein synthesis and transportation via the Golgi complex are required for optimal mMCP-1 secretion. A similar but more marked inhibitory effect with both compounds was demonstrated on the constitutive secretion of CCL2. CONCLUSION: The culture conditions that promote mMCP-1 expression and release by MMC homologues also promote the expression and release of CCL2. Constitutive release involves de novo protein synthesis and requires a functional Golgi complex, suggesting that similar mechanisms of extracellular secretion operate for both mediators. PMID- 12534562 TI - Statutory health assessments for looked-after children: what do they achieve? AB - OBJECTIVE: To examine the outcomes of statutory health assessment of children looked after by an English City Council. DESIGN: Retrospective longitudinal documentary analysis. SETTING AND PARTICIPANTS: Health records of all children looked after by Southampton City Council who had attended at least two statutory health assessments within a designated paediatric service from 1996 to 1999. MAIN VARIABLES STUDIED: Demographic characteristics of the children; physical and mental health problems identified at assessment and extent to which health recommendations were implemented. RESULTS: Twenty-seven boys and 22 girls aged 6 months-15 years were identified who had attended at least two assessments. One hundred and four physical and mental health needs were identified at care entry requiring further assessment or intervention. More health problems were identified for girls than boys. At care entry 15/49 of the children were not fully immunized. At review, on average 14 months later, recommendations had only been implemented in just over half of children. CONCLUSIONS: In common with previous studies this work confirms that the statutory health assessment identifies health need and health neglect that may otherwise go unrecognized. Whereas children's needs and problems were diverse, many continued to suffer health neglect in the system of public care designed to help them. In order to be effective, statutory health assessments must be a health promoting rather than disease screening exercise delivered by professionals skilled to address diverse health needs. Crucially, the heath assessment can only succeed as a tool for health advocacy if complementary to and integrated with local authority care and review. PMID- 12534563 TI - Involving disabled and chronically ill children and young people in health service development. AB - AIM: To investigate the extent and nature of involvement of physically disabled or chronically ill children and young people in local health service development. METHODS: A postal survey of all health authorities (n = 99) and NHS Trusts (n = 410) in England. RESULTS: Seventy-six per cent of health authorities and 59% of Trusts responded. Twenty-seven initiatives involving chronically ill or disabled children and young people in consultation regarding service development were identified. Over half of these were carried out in partnership between health services and other agencies, usually local authorities and/or voluntary organizations. A variety of methods was used for consultation, including child friendly methods such as drawing, drama and making a video. Seventeen initiatives reported that children's involvement had resulted in service changes, but only 11 went beyond consultation to involve children and young people in decision making about service development. Only a third of the organizations had someone with designated responsibility for children's involvement. DISCUSSION: The involvement of this group of children and young people in service development in the NHS is at an early stage. The failure of policy documents on user involvement to identify children and young people as a group for whom methods of consultation need to be developed, and the lack of people with designated responsibility for developing children's involvement may be a reason for slow progress in this area. The initiatives identified show that such involvement is possible and can have a positive impact on services. PMID- 12534564 TI - Development and validation of a tool to measure the impact of childhood disabilities on the lives of children and their families. AB - OBJECTIVE: Information on registers of children with special needs will be more meaningful if a validated measure of the severity of impact of a child's disability on life and family is included. DESIGN: We describe the development and initial validation of a parent-completed questionnaire (Generic Lifestyle Assessment Questionnaire LAQ-G) aimed at measuring such impact. RESULTS: Data were collected on 95 case children, representing various disabilities, and 65 control children without disability, and analysed for case-control, test-re-test and inter-reporter reliability. Multidimensional scaling techniques were then used to derive six domains, representing impact of disability in a structure analogous to the participation domains of the revised International Classification ICF (WHO 2001). CONCLUSIONS: Initial results suggest that the LAQ G is a reliable measure of the impact of disability for children with a range of common disabling conditions. PMID- 12534565 TI - Validation of the measure of processes of care for use when there is no Child Development Centre. AB - INTRODUCTION: Clinical governance requires measurement of the outcomes of health care. The Measure of Processes of Care (MPOC; King, Rosenbaum & King 1995) is a postal questionnaire developed in Canada to reflect parents' perceptions of the quality of services received. AIM: To examine the usefulness of the MPOC for the evaluation of services for children with disabilities and their families. METHODS: The MPOC was revised minimally for British idiom, taking out explicit references to a 'Centre'. Surveys of parents using child disability services were carried out in three rural and two urban communities, where there are no Child Development Centres, with a total of 495 analysable questionnaires returned. In addition, 32 parents using a home nursing service for children with multiple disabilities completed questionnaires. RESULTS: The rate of return ranged from 49% to 67%. Factor analysis confirmed a five-factor solution but only one factor mapped clearly onto the Canadian structure. Using the revised structure, the surveys provide evidence of the discriminating potential of the MPOC, comparing the perceptions of parents who do or do not have a care coordinator, and comparing recipients of a home nursing service with matched users of general services. CONCLUSIONS: The MPOC can be used as a measure of outcome for child disability services of differing organizational structures. PMID- 12534566 TI - Children's experiences of school toilets present a risk to their physical and psychological health. AB - OBJECTIVE: To ascertain why children and parents frequently describe problems with school toilets. SETTING: Two contrasting cities in Northern England (Newcastle upon Tyne 394 pupils) and Southern Sweden (Goteborg/Molndal 157 pupils) METHODS: Self-administered questionnaires were given to children aged 9 11 years in England and Sweden. Researchers administered questionnaires to Head teachers and recorded their observations of facilities according to predetermined basic standards. RESULTS: Children from both countries said they found school toilets unpleasant, dirty, smelly, and frightening and that bullying occurred there. Many children avoided using the school toilets (62% of boys and 35% of girls (in the UK site) and 28% boys and girls in Swedish site avoided using the school toilets to defaecate). Results were similar in both centres. CONCLUSION: European standards are needed for school toilets in order to prevent children developing problems such as constipation, urinary tract infections and incontinence. PMID- 12534567 TI - Cognitive and psychosocial outcome of infants dialysed in infancy. AB - OBJECTIVE: To contribute further to the understanding of cognitive and psychosocial outcome of children with end-stage renal disease undergoing long term peritoneal dialysis. METHODS: In total, 16 surviving infants at a single centre beginning peritoneal dialysis in the first year of life were studied. The age range of the children at assessment was 1.6-12.1 years. Children were assessed using the Griffiths Mental Development Scales, the Wechsler Intelligence Scale for Children-Third Edition UK, and the Strengths and Difficulties Questionnaire. Information regarding the child's hospital stay and family background was also collated. A Pearson's Product Moment correlation was used to analyse the results. RESULTS: Although 67% of the children's scores fell within the average range, 87% were within at least two SDs of the norms (mean IQ = 86.6). Psychosocial adjustment measures revealed that 50% of scores fell within the borderline to abnormal category, suggesting that the frequency of psychological difficulties was above that of the normal population. CONCLUSIONS: These findings lend support to recent studies indicating that, developmentally, children undergoing long-term peritoneal dialysis are faring better than in the past. This may indeed be a reflection of improvements in renal treatment and diet. The behavioural results suggest the need to monitor psychological adjustment in this group of children. PMID- 12534568 TI - Coping with chronic disease from the perspective of children and adolescents--a conceptual framework and its implications for participation. AB - In medical and health psychology, efforts have increasingly been made to assess coping of children and adolescents with chronic conditions. In contrast to the study of coping in adults, approaches to define and assess adaptational processes in children pose a number of problems because coping and development are inherently connected with each other. Issues arising when applying theoretical concepts from developmental psychology to the area of coping in children and adolescents are highlighted. The most prominent approaches to conceptualize and assess coping with chronic disease in childhood and adolescence are illustrated. In future research, there is a need to focus the situational context and content of coping rather than to assess the effort employed and level of a particular coping strategy. Coping is not only a way of regulating emotions, but has an interpersonal meaning, depending on its interactional context. In the medical field, coping has a mediating function for participation and shared medical decision-making in health care processes. PMID- 12534569 TI - Parents' stories of sensitive and insensitive care by paediatricians in the time leading up to and including diagnostic disclosure of a life-limiting condition in their child. AB - This study explored parents' experiences of care by paediatricians in the time leading up to and including diagnostic disclosure of a life-limiting condition in their child. RESEARCH METHODS: Qualitative exploration using in depth interviews with a purposive sample of 30 families whose child was diagnosed with a life limiting condition. RESULTS: Parents' stories centred upon their need to have their concerns and needs as parents validated by paediatricians. Demonstrable qualitative differences were shown to exist between the sensitive and insensitive clinical practices of paediatricians. Parents' stories identify the need for clinical practice to be based upon a humanistic approach to care. These also identify how sensitive paediatricians achieved this ideal through meeting the 'double obligation' of combining technical skills with an understanding of their needs as parents. PMID- 12534572 TI - A review of intractable facial pain secondary to underlying lung neoplasms. AB - We describe a 63-year-old smoker who suffered from intractable facial pain secondary to an underlying lung neoplasm. Data from 30 previously reported and similar cases are also summarized. The clinical triad of a smoker suffering from periauricular pain and an elevated ESR should alert the clinician to the possibility of an occult lung mass. In these cases a computed tomography of the chest should always be obtained. Previously refractory pain typically responds to surgical resection of the mass and/or radiation therapy. PMID- 12534573 TI - A community-based study of headache diagnosis and prevalence in Singapore. AB - We present the results of a community survey based on the diagnostic criteria of the International Headache Society (IHS) describing headache prevalence and symptomatology in the Singapore population. A questionnaire administered by trained personnel was completed by 2096 individuals from a randomized sample of 1400 households. The overall lifetime prevalence of headache was 82.7%. The migraine prevalence was 2.4% in males and 3.6% in females; for episodic tension type headache and chronic tension-type headache the corresponding figures were 11.1%/11.8% and 0.9%/1.8%, respectively. Inclusion of borderline cases (IHS codes 1.7 and 2.3) resulted in prevalences of 9.3% for migraine, 39.9% for episodic tension headache and 2.4% for chronic tension headache. Headaches described by 31.2% of the respondents were unclassifiable. The different premonitory symptoms, precipitants and aggravating factors in migraine and tension-type headache in our study population suggest that they represent two distinct syndromes rather than opposite ends of a clinical spectrum. PMID- 12534574 TI - Increased frequency of migraine in narcoleptic patients: a confirmatory study. AB - Previously we have reported an increased prevalence of migraine in narcoleptic patients. Because of the theoretical and clinical implications of this finding we recruited an independent new study sample of 100 patients with proven narcolepsy and conducted a structured 26-item interview based on the international diagnostic criteria for headache disorders, the Kiel Headache Questionnaire. Narcolepsy symptoms were measured by means of the Stanford Centre for Narcolepsy Sleep Inventory. Migraine prevalence was twofold to fourfold increased in the narcoleptic patients and amounted to 44.4% in women and 28.3% in men. The onset of narcolepsy symptoms was 12.3 +/- 11.4 years before the onset of migraine symptoms. The results might be regarded as indicative of a common pathophysiological pathway relevant to both of the two disorders. PMID- 12534575 TI - Hypnic headache - the first German cases including polysomnography. AB - We report the first four German cases of hypnic headache according to the criteria suggested in the literature. Furthermore, we present the results of polysomnography in two theses cases with hypnic headache occurring during REM sleep. In one case, hypnic headache was also associated with periodic limb movements. Although mean nocturnal oxygen saturation was decreased in another patient, nightly oxygen inhalation did not result in an improvement of the headache. PMID- 12534576 TI - The paroxysmal hemicrania-tic syndrome. AB - Two cases of paroxysmal hemicrania (PH) associated with trigeminal neuralgia are reviewed. The paroxysmal hemicrania component in one patient was episodic, while it was chronic in the other. Each headache type responded completely to separate treatment, highlighting the importance of recognizing this association. We review the six other cases of chronic paroxysmal hemicrania-tic (CPH-tic) reported, and suggest that the term paroxysmal hemicrania-tic syndrome (PH-tic) be used to describe this association. PMID- 12534577 TI - Central 5-HT receptor hypersensitivity in migraine without aura. AB - Serotonin has long been implicated as a key neurotransmitter in migraine. There is a dearth of research specifically examining 5-HT1A receptor sensitivity in migraine despite the importance of this receptor in regulating central serotonergic tone. In this study we examined the hypothesis that migraine without aura is associated with hypersensitivity of central 5-HT1A receptors, using a 5 HT1A neuroendocrine challenge drug and comparing serum prolactin responses between a test group with migraine and a matched group of healthy controls. Twelve female subjects fulfilling International Headache Society (IHS) criteria for migraine without aura were evaluated. Following an overnight fast, subjects presented for testing at 9am. An intravenous canula was inserted and serum prolactin was assessed at baseline and every 30 min for 3 h following a single dose of 30 mg oral buspirone, a 5-HT1A-receptor agonist. Subjects were assessed during the first 5 days of the menstrual cycle. No subjects were taking psychotropic medication or migraine prophylactic treatment. Patients with current or previous psychiatric disorder, daily headache or analgesic overuse were excluded. 16 healthy female volunteers matched for age and menstrual status were also evaluated and served as controls. There was no difference in baseline prolactin between groups. There was a significant rise in prolactin following buspirone in both groups. Subjects with migraine had a significantly increased prolactin response to buspirone (delta max) compared to controls (P < 0.001). This study supports the hypothesis that migraine without aura is associated with a relative hypersensitivity of central 5-HT1A receptors. This is of relevance given the role of the 5-HT1A receptor in controlling raphe 5-HT tone and in the possible association between migraine and anxiety and depression. PMID- 12534578 TI - Trigemino-cervical reflex in patients with headache. AB - Neurophysiological studies have shown abnormal activity of some brainstem nuclei in headache patients. The trigemino-cervical reflex is an anti-nociceptive reflex that gives an opportunity for evaluation of the brainstem interneurone activity. It has not been previously examined in headache patients. We studied 15 patients with predominantly unilateral chronic tension-type headache, 15 patients with migraine without aura and 32 healthy subjects. The trigemino-cervical reflex was recorded bilaterally from the resting sternocleidomastoid muscle using surface electromyographic recordings. In all headache patients the trigemino-cervical reflex on the painful side was with shortened latency compared with the non painful side and with healthy persons. The results suggest decreased activity of the brainstem inhibitory interneurones. We suggest that although the pathophysiological mechanisms of tension-type headache and migraine are different, they share common mechanisms of abnormal pain control. PMID- 12534579 TI - Urinary nitric oxide metabolites and lipid peroxidation by-products in migraine. AB - Enhanced endothelium nitric oxide (NO) and superoxide anion release may cause migraine through related cerebral blood flow changes. Thirty subjects suffering from migraine with and without aura and 20 healthy controls were investigated. Urine samples collected for 24 h during and after the migraine attack, and during the headache-free period, were assayed for urinary NO stable metabolites (NOx) and thiobarbituric acid reactive substances (TBARS). During the headache-free period urinary NOx and TBARS levels were higher in migraine sufferers than in controls (NOx 0.77 +/- 0.14 vs. 0.28 +/- 0.15 mmol/mmol creatinine, P < 0.05; TBARS 0.40 +/- 0.19 vs. 0.26 +/- 0.13 micro mol/mol creatinine, P < 0.05). Also, NOx excretion was higher during the headache-free period than during or after the migraine attack (P < 0.05). Urinary TBARS were increased during the attack with respect to the headache-free period (P < 0.05). No differences were observed in the same parameters between sufferers of migraine with and without aura. Urinary NOx and TBARS might be promising as markers of their systemic levels to evaluate the increased vulnerability to oxidative stress in migraine sufferers. PMID- 12534580 TI - 'Side locked' migraine and trigeminal autonomic cephalgias: evidence for clinical overlap. AB - This paper will discuss evidence which supports a link between 'side locked' migraine (SLM) and the trigeminal autonomic cephalgias (TACs). Recent papers brought strictly unilateral primary headaches into focus, proposing new classification and discussing pathophysiological mechanisms. We reviewed those proposals and present evidence that SLM falls in between the well-defined TACs and side shifting migraine (SSM). It is difficult to differentiate SLM from the recently proposed headache subtype called hemicrania generis incerti (i.e. hemicrania continua unresponsive to indomethacin). We also present cases that may exemplify the considerations made in the paper. PMID- 12534581 TI - Extracephalic jabs/idiopathic stabs. Vaga study of headache epidemiology. AB - Ultrashort cephalic paroxysms are well known. In the parish of Vaga, Norway, 35.2% of the 18-65-year-old subjects (n = 1779) were recently found to have such jabs. In the present work, a search has been made for extracephalic 'jabs'. A questionnaire was in its entirety administered by the same investigator (O.S.) in a 'semistructured' way. Facial jabs were present in three women, and in one of them the pain spread to the head. Four subjects had jabs occurring at random throughout the body, also including the cephalic area. Pure nuchal jabs were present in 12 subjects, 10 of whom were males. This sex preponderance difference differs significantly from that in jabs in general (with 40.2% males). The characteristics of the extracephalic jabs, i.e. the duration and temporal pattern, do not seem to differentiate them essentially from jabs in general. The subjects were not asked specific questions regarding extracranial jabs. Most of the affected individuals gave information spontaneously about their jabs. For these reasons, this study is not a proper prevalence study. It does show, however, that extracranial jabs exist, and it gives some indications as to their frequency. PMID- 12534582 TI - Decreased sTNF-RI in migraine patients? AB - Tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) have recently been found to have a pain-mediating function in addition to their immunological, proinflammatory function. According to the hypothesis of neurovascular inflammation in migraine, these two cytokines could contribute to migraine pain generation. We analysed IL-6 and its soluble receptors sIL-6R and sgp130 as well as TNF-alpha and its soluble receptor sTNF-RI in 27 migraine patients and eight headache-free controls. Migraine patients tended to have less sTNF-RI (794 +/- 158 pg/ml) than controls (945 +/- 137 pg/ml). No differences in cytokine concentrations were observed. If TNF-alpha plays a role in migraine physiopathology, migraine patients may lack sufficient antagonistic sTNF-RI to neutralize hyperalgesic TNF-alpha during a migraine attack. PMID- 12534583 TI - Aspirin in episodic tension-type headache: placebo-controlled dose-ranging comparison with paracetamol. AB - Most people with episodic tension-type headache (TTH) treat themselves with over the-counter analgesics. In the absence of clear evidence of dose-related efficacy of the two most commonly used analgesics, aspirin (acetylsalicylic acid) and paracetamol (acetaminophen), this study compared two doses of each with placebo. In a double-blind, double-dummy, randomized parallel-groups comparative trial, 638 consenting subjects aged 16-65 years with episodic TTH (but not migraine) by IHS criteria were recruited from the UK general population by advertisement. They treated one episode of moderate or severe TTH with a single dose of 500 or 1000 mg aspirin, 500 or 1000 mg paracetamol or placebo. The primary objective was to compare aspirin 1000 mg with placebo, and the primary end-point was subjective pain relief (total or worthwhile) 2 h after treatment ('response'). Additionally, pain intensity on a 100-mm visual analogue scale and functional impairment were monitored regularly for 4 h and at 24 h, although rescue medication was allowed after 2 h. The analysis was of the intention-to-treat population of 542 who took treatment (all providing outcome data). Treatment groups were matched at baseline. Aspirin 1000 mg (75.7% response rate; P = 0.0009) and to a lesser extent aspirin 500 mg (70.3%; P = 0.011) and paracetamol 1000 mg (71.2%; P = 0.007), but not paracetamol 500 mg (63.8%; P = 0.104), were statistically more effective than placebo despite a high placebo-response rate (54.5%). Outcome was not affected by headache intensity at baseline. Secondary end-points including functional recovery (by median times of 4.0-13.5 h) were consistent with these findings, although a minority of subjects recorded long-duration functional impairment (37-54 h). Adverse events reported by 13.4-18.9% of subjects were mild or moderate, and transient. No safety concerns arose. PMID- 12534584 TI - Autonomic activation in experimental trigeminal pain. PMID- 12534585 TI - Treatment of cluster headache with topiramate: effects and side-effects in five patients. PMID- 12534588 TI - The molecular genetics of the genodermatoses: progress to date and future directions. AB - The Human Genome Mapping Project and allied rapid advances in genetic technology over the past decade have facilitated accurate association of allelic variations in several genes with specific skin phenotypes. Currently the genetic bases of the majority of the more common genodermatoses have been elucidated. In scientific terms this work has been extraordinarily successful and has yielded many new biological insights. These advances, although exciting, have yet to be translated into direct benefit for patients with these diseases. Genetic counselling has been greatly aided by gene identification, by the better understanding of genotype-phenotype correlation and by the disclosure of unexpected genetic mechanisms in some families. Knowledge of the molecular basis of these disorders has also been vital in enabling DNA-based prenatal diagnosis in several conditions and DNA-based preimplantation diagnosis has been used in a selected few. While this successful period of gene mapping is now nearing completion, progress towards the next goal, that of developing therapeutic strategies based on the knowledge of these underlying genetic mechanisms, has proven frustratingly slow. Despite the ready access to the skin compared with solid internal organs, the challenges of cutaneous gene therapy are legion and many technical issues need to be surmounted to enable gene replacement or modification of gene expression to have a useful role in these disorders. In this article we make a comprehensive review of progress to date in gene identification, genotype-phenotype correlation, prenatal diagnosis and cutaneous gene therapy, and we examine future directions for research in this field. PMID- 12534589 TI - Degradation of extracellular matrix components by defined proteinases from the greenbottle larva Lucilia sericata used for the clinical debridement of non healing wounds. AB - BACKGROUND: Larvae of the greenbottle fly Lucilia sericata are used routinely for the clinical treatment of difficult necrotic and infected wounds. Degradation by proteinases contained in larval excretory/secretory (ES) products is thought to contribute to wound debridement by removal of dead tissue. However, proteinase activity may also affect host tissue remodelling processes. OBJECTIVES: To identify proteolytic enzymes derived from L. sericata ES products with activities against fibrin and extracellular matrix (ECM) components. METHODS: Larval proteinase activities were assayed in vitro using class-specific substrates and inhibitors. Their action against fibrin and ECM components was examined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis. RESULTS: Three classes of proteolytic enzyme were detected in the secretions using fluorescein isothiocyanate-labelled casein as a model substrate. The predominant activity belonged to serine proteinases (pH optima 8-9) of two different subclasses (trypsin-like and chymotrypsin-like), with a weaker aspartyl proteinase (pH 5) and a metalloproteinase (pH 9) with exopeptidase characteristics also present. Using skin-relevant ECM components as substrates L. sericata ES products solubilized fibrin clots and degraded fibronectin, laminin and acid-solubilized collagen types I and III. Hydrolysis of ECM macromolecules was inhibited by preincubating ES products with phenylmethylsulphonyl fluoride but not 4 amidinophenylmethylsulphonyl fluoride, indicating that degradation was due to the 'chymotrypsin-like' serine proteinase. CONCLUSIONS: These data suggest that a combination of L. sericata ES proteinases involving chymotrypsin-like and trypsin like activities could potentially influence wound healing events when maggots are introduced into necrotic and infected wounds, with the chymotrypsin-like activity involved in the remodelling of ECM components. PMID- 12534590 TI - Isolation of a CD8alphaalpha+ CD4- tumour T-cell clone with cytotoxic activity from a CD4+ CD8- cutaneous T-cell lymphoma. AB - BACKGROUND: We have previously established tumour T-cell lines, both from the skin and from the blood of patients with a cutaneous T-cell lymphoma (CTCL). In one patient, the tumour cells and the derived cell lines had a CD3+ CD4+ CD8- phenotype and a trisomy of chromosome 7. They expressed three T-cell receptor (TCR) beta-chain transcripts, but only one was productively rearranged and expressed at the cell membrane. OBJECTIVES: In the present study, we tried to isolate a fast-growing new tumour T-cell line from the same patient. PATIENTS/METHODS: We performed direct cell cloning of the skin tumour lymphocyte population, which led to the isolation of an interleukin-2-dependent highly proliferative T-cell subclone, named Cou-L3, with a CD3+ TCR-Vbeta13+ CD4- CD8alphaalpha+ phenotype. RESULTS: We demonstrated that Cou-L3 was identical to the original clonal tumour CD3+ Vbeta13+ CD4+ CD8- cells, as it expressed the same rearranged TCR-Vbeta13 chain. We further studied the functional activity of these CD8alphaalpha+ Vbeta13+ Cou-L3 cells. We found that these cells exhibited CD3-redirected cytotoxic activity. CONCLUSIONS: An immunophenotypic shift, with a change from a CD4+ to a CD8+ phenotype, has been already reported in association with disease progression in CTCL. However, in these cases, there has been no demonstration that the phenotypic change involved the same T-cell clone. The present study is the first report of the phenotypic heterogeneity of the tumour clonal cell population in CTCL. PMID- 12534591 TI - Adrenomedullin: expression and possible role in human skin and hair growth. AB - BACKGROUND: Adrenomedullin (AM) is a regulatory peptide that is synthesized and secreted by a wide number of cells and tissues. AM is a potent vasodilator, but also exerts other functions, such as regulating cell growth and antimicrobial defence. Two receptors, L1 and calcitonin receptor-like receptor (CRLR), which are able to bind AM, have been cloned and characterized. OBJECTIVES: To investigate expression of AM protein and its receptors in human skin and during different stages of the human hair cycle and, moreover, because of the suggested antimicrobial function of AM in skin, to investigate AM immunoreactivity (IR) in inflammatory acne lesions compared with healthy pilosebaceous follicles. METHODS: We used immunohistochemistry to determine the distribution of AM and its receptors in human skin and during different stages of the human hair cycle. AM IR in inflammatory acne lesions was investigated to evaluate the antimicrobial function of the protein, and hair follicle cultures were performed to examine the role of AM in differentiation and proliferation of hair follicle keratinocytes. RESULTS: Strong IR for AM and its receptors was present in the suprabasal epidermis, in the melanocytes of the epidermis, and in sweat and sebaceous glands. In the hair follicle, AM protein was strongly expressed in the basal and suprabasal layers of the hair bulb and the proximal outer root sheath (ORS). In the distal ORS, AM expression was increasingly suprabasal, especially in proximity to the bulge region where the basal cell layer was free of IR. IR for the CRLR revealed a similar expression pattern to that seen for AM. In contrast, L1 IR showed a suprabasal pattern of IR throughout the ORS. Similar expression of AM and its receptors was observed in catagen and early anagen follicles. AM expression was not markedly upregulated in acne lesions, suggesting a minor role for this antimicrobial peptide in acne. Despite its well-documented mitogenic effects, particularly in oral and skin keratinocytes, AM had no significant effect on hair follicle growth in vitro. CONCLUSIONS: AM and its receptors are expressed in human hair follicles, and both AM and its receptors are colocalized in the same compartments and cell types of the skin. This finding is consistent with the proposed autocrine/paracrine mechanism in the physiology of AM. PMID- 12534592 TI - Modulation of collagen synthesis and mRNA by continuous and intermittent use of topical hydrocortisone in human skin. AB - BACKGROUND: Glucocorticoids have been shown to downregulate collagen synthesis in human skin in vivo, thereby contributing to skin atrophy. OBJECTIVES: To compare the effects of continuous and intermittent use of topical hydrocortisone on skin collagen synthesis and, furthermore, to elucidate the mechanism of collagen synthesis reduction induced by hydrocortisone. METHODS: Collagen propeptides reflecting the synthesis rate of type I and III collagens were studied from suction blister fluids in nine healthy subjects after 3 weeks of continuous (twice daily) or intermittent (on three consecutive days weekly) topical hydrocortisone treatment and 2 weeks after the termination of treatment. Type I collagen mRNA was studied in the same subjects from skin biopsies by using in situ hybridization (ISH) after 3 weeks of treatment. RESULTS: Three weeks of continuous treatment decreased the types I and III collagen propeptides in suction blister fluid by 89% and 82%, respectively, while intermittent treatment resulted in a corresponding decrease of 53% and 50%. ISH studies from skin biopsies showed type I collagen mRNA to be markedly reduced in fibroblasts after continuous and intermittent steroid treatment. After a 2-week drug-free interval, the synthesis rate was completely restored in both areas, and some subjects even showed upregulation of synthesis in previously steroid-treated skin. CONCLUSIONS: Continuous hydrocortisone for 3 weeks markedly decreases collagen propeptides and corresponding mRNA in human skin. Intermittent hydrocortisone has a less marked effect on the collagen synthesis rate. PMID- 12534593 TI - Effects of staphylococci on cytokine production from human keratinocytes. AB - BACKGROUND: Staphylococcus skin infection is characterized by the infiltration of numerous neutrophils within the epidermis; however, the precise mechanism of epidermal infiltration of neutrophils during skin infection with staphylococci is not well understood and the factors regulating the neutrophil recruitment are yet to be determined. OBJECTIVES: We investigated the effects of staphylococci on cytokine production from keratinocytes, specifically to elucidate the mechanisms of neutrophil infiltration within the epidermis in cutaneous microbial infection. METHODS: Cytokine production from human keratinocytes was examined after stimulation with heat-killed Staphylococcus aureus, S. epidermidis and S. intermedius. RESULTS: Interleukin (IL)-6 and IL-8 were detected in the culture supernatants by enzyme-linked immunosorbent assay but IL-1beta, monocyte chemotactic protein-1 and tumour necrosis factor-alpha were not. IL-6 and IL-8 mRNAs were also confirmed by reverse transcription-polymerase chain reaction in the keratinocytes stimulated with killed staphylococci for 1, 3, 6, 10 and 24 h. CONCLUSIONS: These results could explain the epidermal infiltration of neutrophils in cutaneous infection with staphylococci, suggesting that the analysis of cytokines might add valuable information for the pathogenesis of cutaneous infection with Staphylococcus species. PMID- 12534594 TI - Molecular strain typing of Trichophyton rubrum indicates multiple strain involvement in onychomycosis. AB - BACKGROUND: Trichophyton rubrum is an important cause of onychomycosis. Molecular strain typing methods have recently been developed to address questions of epidemiology and source of relapse following treatment. OBJECTIVES: To determine whether T. rubrum nail infections are caused by one or more strains of this fungus. METHODS: Nail specimens from 10 patients with onychomycosis due to T. rubrum were cultured and five colonies per culture plate were selected for molecular strain typing. DNA was extracted from these isolates and subjected to a polymerase chain reaction-based typing method that analyses variations in numbers of repetitive elements in the non-transcribed spacer region of the ribosomal RNA gene repeats. RESULTS: In six of 10 specimens, there were two or more T. rubrum strain types present. CONCLUSIONS: This preliminary study suggests that in many cases of fungal nail infection by T. rubrum, multiple strains are involved. This has important implications for epidemiological studies and possibly for therapy. PMID- 12534595 TI - Chromosomal abnormalities in relation to clinical disease in patients with cutaneous T-cell lymphoma: a 5-year follow-up study. AB - BACKGROUND: Patients with cutaneous T-cell lymphoma (CTCL) show chromosomal aberrations in skin and blood lymphocytes. OBJECTIVES: To evaluate the significance of peripheral blood clonal or non-clonal chromosomal abnormalities in comparison with the clinical course of cutaneous T-cell lymphoma patients. PATIENTS/METHODS: Five patients with large-plaque parapsoriasis (LPP) or with follicular mucinosis, eight with mycosis fungoides and two with Sezary syndrome were followed for an average of 54 months. G-banding and enzyme-detected in situ hybridization (EDISH) were used to identify aberrations in chromosomes 1, 6, 8, 9, 11, 13/21, 15 or 17, that had previously showed frequent aberrations. RESULTS: The aberration rates of all chromosomes studied differed between patients with active disease and healthy or photochemotherapy-treated controls by EDISH or G banding (P < 0.01 to P < 0.05). Patients in complete remission differed from healthy controls for aberrations of chromosomes 1, 6 and 11, and from patients with active, progressing disease for chromosomes 1, 6, 8, 11 and 17 (P < 0.01 to P < 0.05, EDISH or G-banding). All 11 samples representing active, progressing disease showed elevated levels of chromosome 8 aberrations in EDISH. The change in chromosomal aberration rate and clinical condition between two consecutive samples agreed for chromosomes 1, 8, 9 and 15 (G-banding) and for chromosome 17 (G-banding and EDISH; kappa > 0.5-0.6). Six of seven patients (five CTCL, one LPP patient) with clonal chromosomal aberrations by G-banding showed continuously active disease and four of them, but none of the other patients, died within 30 months of the detection of the clone. CONCLUSIONS: The rate of chromosomal aberrations associates with the activity of CTCL, and has prognostic significance. Aberrations of chromosomes 1, 6 and 11, although increasing with activity of the disease, seem to be a hallmark of existing disease, detectable even in remission. Aberrations of chromosomes 8 and 17 especially associate with active or progressive disease. PMID- 12534596 TI - Effects of skin occlusion in patch testing with sodium lauryl sulphate. AB - BACKGROUND: When evaluating transepidermal water loss (TEWL) in patch testing, the occlusive effect of the patch must be considered as an important artificial impairment of the measurement. OBJECTIVES: To investigate the time course of effects of occlusion. METHODS: Epicutaneous patches with sodium lauryl sulphate (SLS) 0.25%, SLS 0.5%, water and an empty test chamber (control) were applied on the volar forearm for different time intervals (12, 24, 48 h). Test reactions were evaluated by measurement of TEWL immediately, every 15 min during the first hour, every 30 min during the following 3 h and 24 h after patch removal. RESULTS: After patch removal, TEWL values showed a steep increase. When compared with basal values, TEWL values after SLS patch testing remained increased for 24 h, whereas TEWL values on water patch sites were only significantly increased for up to 180 min, and on empty patch sites for only up to 120 min after patch removal. The prolonged increase in TEWL values in SLS patch testing seemed to be induced by barrier function damage caused by SLS itself, as shown in various earlier studies. After the initial increase, TEWL values showed a significant decrease for all patches from 0 to 120 min after patch removal. Patch testing with water gave a significant decrease in TEWL values up to 180 min, and for empty chambers (control) up to 150 min after removal of patches. These data suggest that the occlusive effect on TEWL in patch testing ends 3 h after the removal of test chambers. CONCLUSIONS: We recommend TEWL measurement in SLS patch testing after a period of at least 3 h after patch removal. For practical purposes a 24-h period after patch removal may be useful. PMID- 12534597 TI - Oral lichenoid reactions associated with amalgam: improvement after amalgam removal. AB - BACKGROUND: The pathogenetic relationship between oral lichenoid reactions (OLR) and dental amalgam fillings is still a matter of controversy. OBJECTIVES: To determine the diagnostic value of patch tests with amalgam and inorganic mercury (INM) and the effect of amalgam removal in OLR associated with amalgam fillings. METHODS: In 134 consecutive patients 467 OLR were classified according to clinical criteria. One hundred and fifty-nine biopsies from OLR lesions were histologically diagnosed according to the World Health Organization criteria for oral lichen planus (OLP) and compared with 47 OLP lesions from edentulous patients without amalgam exposure. One hundred and nineteen patients were patch tested with an amalgam series. In 105 patients (357 of 467 lesions) the amalgam fillings were removed regardless of the patch test results and OLR were re examined within a follow-up period of about 3 years. Twenty-nine patients refused amalgam removal and were taken as a control group. RESULTS: Eleven patients with OLR (8.2%) had skin lesions of lichen planus (LP). Histologically, the lesions in the OLR group could not be distinguished from those seen in the OLP group. Thirty three patients (27.7%) showed a positive patch test to INM or amalgam. Amalgam removal led to benefit in 102 of 105 patients (97.1%), of whom 31 (29.5%) were cured completely. Of 357 lesions, 213 (59.7%) cleared after removal of amalgam, whereas 65 (18.2%) did not improve. In the control group without amalgam removal (n = 29) only two patients (6.9%) showed an improvement (P < 0.05). Amalgam removal had the strongest impact on lesions of the tongue compared with lesions at other sites (P < 0.05), but had very little impact on intraoral lesions in patients with cutaneous LP compared with patients without cutaneous lesions (P < 0.05). Patients with a positive patch test reaction to amalgam showed complete healing more frequently than the amalgam-negative group (P < 0.05). After an initial cure following amalgam removal, 13 lesions (3.6%) in eight patients (7.6%) recurred after a mean of 14.6 months. CONCLUSIONS: Of all patients with OLR associated with dental amalgam fillings, 97.1% benefited from amalgam removal regardless of patch test results with amalgam or INM. We suggest that the removal of amalgam fillings can be recommended in all patients with symptomatic OLR associated with amalgam fillings if no cutaneous LP is present. PMID- 12534598 TI - Pulsed dye laser treatment of telangiectasia after radiotherapy for carcinoma of the breast. AB - BACKGROUND: Chronic radiodermatitis after radiotherapy for carcinoma of the breast is a common sequela of treatment and can be distressing for the patient. The skin is atrophic and shows prominent telangiectasia due to dilatation of a reduced or poorly supported skin vasculature. The pulsed dye laser (PDL) is an established treatment of cutaneous telangiectatic disorders including facial telangiectasia and spider naevi, and is safe and efficacious. OBJECTIVES: To study the efficacy of the PDL in the treatment of postradiation telangiectasia of the breast or chest wall. METHODS: Prospective open study of the treatment of eight females with the Candela SPTL1B PDL. Subjective assessments of vessel clearance, adverse effects and patient questionnaires. RESULTS: All treated patients showed complete clearance of vessels. Two patients developed hypopigmentation. All patients reported a high degree of satisfaction with the treatment. CONCLUSIONS: PDL therapy clears postirradiation telangiectasia of the breast and chest wall successfully with minimal adverse reactions, and can be recommended for patients distressed by this disorder. PMID- 12534599 TI - Treatment of acquired junctional melanocytic naevi by Q-switched and normal mode ruby laser. AB - BACKGROUND: Acquired junctional melanocytic naevi are harmless pigmented lesions of the epidermis, which can be of cosmetic concern. Various therapeutic approaches have been used in the treatment, but all these methods produce postoperative scarring or alterations in skin texture. Pigment laser treatment of benign pigmented lesions has shown a low potential for scarring by selectively targeting melanosomes in melanocytes and keratinocytes. OBJECTIVE: To find a fast, effective and safe treatment for the removal of acquired junctional melanocytic naevi. PATIENTS/METHODS: We first studied the effect of the Q switched and normal mode ruby laser on 12 patients (eight women and four men) with acquired melanocytic naevi. The effect was monitored by histology and clinical photography. RESULTS: If the response to one treatment with the Q switched laser mode was not completely effective, the lesions were subsequently treated with one or two sessions with the laser in normal mode. All flat lesions responded completely. After a follow-up period of 1 year they had not recurred. Slightly elevated lesions showed only a partial response, e.g. disappearance of the junctional part of the naevus but recurrence of the dermal part of the naevus. Red-brown junctional naevi as seen in skin types I and II did not respond well to ruby laser treatment. CONCLUSIONS: The Q-switched ruby laser was very successful in completely removing flat (non-palpable) acquired junctional melanocytic naevi, but not compound naevi, with one to three treatment sessions, without any scarring or pigmentary disturbance. PMID- 12534600 TI - Hyaluronan, CD44 and versican in epidermal keratinocyte tumours. AB - BACKGROUND: The high molecular weight polysaccharide hyaluronan is a major component of the extracellular matrix between the vital cells of human skin epidermis. The levels of hyaluronan, and those of the hyaluronan receptor CD44 and the hyaluronan binding proteoglycan versican, correlate with the aggressiveness of different human carcinomas of epithelial origin. OBJECTIVES: To study skin keratinocyte tumours for the expression of hyaluronan, the hyaluronan receptor CD44 and the hyaluronan binding proteoglycan versican. METHODS: Paraffin embedded sections of 114 basal cell carcinomas (BCC), 31 in situ carcinomas (ISC) and 35 squamous cell carcinomas (SCC) were stained with a hyaluronan specific probe, biotinylated hyaluronan binding complex, and with monoclonal antibodies against CD44 and versican. RESULTS: Compared with normal epidermis, ISC and well differentiated SCCs showed an enhanced hyaluronan signal on carcinoma cells while CD44 expression level resembled that of normal skin. Less differentiated SCCs showed reduced and irregular expression of both hyaluronan and CD44 on carcinoma cells. In BCCs, hyaluronan and CD44 signals were absent or very low on the surface of carcinoma cells. However, hyaluronan was frequently present on BCC cell nuclei, a feature completely absent in ISC, SCC and normal epidermis. An accumulation of hyaluronan in the connective tissue stroma around the tumour was more frequent in SCCs than BCCs. Versican staining was positive around hair follicles and dermal blood vessels of normal skin. Peritumoral versican signal was present in a part of the BCCs but not in other tumours. CONCLUSIONS: The completely different hyaluronan and CD44 expression patterns in BCC and SCC probably reflect the different origins of the tumours, with BCC an undifferentiated keratinocyte and SCC a keratinocyte at an early stage in the differentiation pathway. The difference in hyaluronan and CD44 expression between these tumours may also contribute to the difference in their capacity to metastasize. PMID- 12534601 TI - Melanocyte detachment after skin friction in non lesional skin of patients with generalized vitiligo. AB - BACKGROUND: In vitiligo, melanocytes are gradually lost in depigmented macules of the skin. The disappearance of melanocytes has, however, not been clearly observed and consequently the aetiology of the disease (autoimmune, neural, cytotoxic) is still elusive. The starting point of vitiligo macules is frequently determined by local conditions such as wounds and excoriations, but may also follow minor traumas such as pressure or repeated friction. This prominent feature is often neglected. OBJECTIVES: To clarify the biological consequences of repeated friction on the attachment and survival of melanocytes in non lesional vitiligo skin. METHODS: Light reproducible skin friction was performed for 4 min on the volar forearm of 18 patients with extensive vitiligo and five controls with normal healthy skin. Biopsies from the test area and control skin were taken at 1, 4, 24 and 48 h following friction. Serial sections were examined with standard light microscopy, transmission electron microscopy, histochemistry and immunohistochemistry (dihydroxyphenylalanine, HMB-45, E-cadherin and an early apoptosis marker, M30 cytoDEATH antibody). RESULTS: The observation of sections at 1 and 48 h after friction on vitiligo skin and at all time points in controls revealed no changes. In contrast, in vitiligo skin at 4 and 24 h after friction, several melanocytes had undergone detachment and were found in various suprabasal positions, including the stratum spinosum, granular layer, and within and outside the stratum corneum. CONCLUSIONS: Detachment and transepidermal elimination of melanocytes following minor mechanical trauma in non lesional vitiligo skin is probably the cause of depigmentation occurring in the isomorphic response (Koebner phenomenon). We propose that transepidermal elimination of melanocytes in vitiligo should be regarded as a possible mechanism of chronic loss of pigment cells, perhaps previously damaged by another process. PMID- 12534602 TI - Lamin expression in normal human skin, actinic keratosis, squamous cell carcinoma and basal cell carcinoma. AB - BACKGROUND: Aberrant expression patterns of nuclear lamins have been described in various types of cancer depending on the subtype of cancer, its aggressiveness, proliferative capacity and degree of differentiation. In general, the expression of A-type lamins (lamins A and C) has been correlated with a non-proliferating, differentiated state of cells and tissues. OBJECTIVES: To establish and compare the expression patterns of lamins in normal human skin, actinic keratosis (AK), squamous cell carcinoma (SCC) and basal cell carcinoma (BCC). METHODS: Expression patterns of the individual lamin subtypes were studied immunohistochemically. The proliferation capacity of the tumour cells was detected using a specific antibody to Ki-67, and was related to the A-type lamin expression patterns. RESULTS: In normal skin, lamin A was expressed in the suprabasal cell compartment of the epidermis, whereas the basal cells were mostly unstained. BCCs and SCCs stained positive in most cells, while the epidermis overlying BCC and SCC and the epidermis in AK stained homogeneously and strongly in the basal cells in addition to the suprabasal cells. Lamin C was expressed in some basal cells of normal epidermis while the suprabasal cells stained strongly positive. Both BCCs and SCCs stained strongly positive for lamin C, with the difference that in BCC the staining was predominantly present in nucleolar structures with occasional staining of the nuclear envelope. The epidermis overlying SCC showed strong positivity in the lamina of virtually all cells. The expression of lamin C in the basal cells of AK resembled the expression pattern seen in the epidermis overlying BCC, i.e. a nucleolar staining next to nuclear envelope staining. Lamin B1 and B2 were found in virtually all cells in normal epidermis, AK, BCC, SCC and the epidermis overlying cancer. The percentage of Ki-67-expressing cells was highest in BCC (45%), and gradually decreased via epidermis overlying BCC, AK, SCC, and epidermis overlying SCC, to normal skin (11%). Simultaneous expression of A-type lamins and Ki-67 occurred in approximately 50% of the proliferating (Ki 67 positive) cells in BCC and SCC. CONCLUSIONS: Significant changes occur in the expression patterns of A-type lamins in both premalignant and malignant lesions of the skin. The profound overlap of lamin A and Ki-67 staining patterns indicates that the proliferating tumour cells may obtain a certain degree of differentiation. Finally, lamin A expression in the basal cell layer of the apparently normal epidermis overlying BCC may suggest its involvement in the primary process. PMID- 12534603 TI - Gender differences in rising trends in cutaneous malignant melanoma in Spain, 1975-98. AB - BACKGROUND: Cutaneous malignant melanoma (CMM) morbidity and mortality rates have increased over the past several decades all around the world, but some developed countries have experienced recent declines in mortality from CMM among the young and middle-aged and even a reversal in the long-term increase in deaths attributable to CMM. OBJECTIVES: To describe CMM mortality trends, by age and gender, in Spain between 1975 and 1998. METHODS: Crude, age-adjusted, truncated, cumulative, age-specific and potential years of life lost rates of CMM mortality were calculated by gender. Age and period Poisson regression models were fitted to gender- and age-specific CMM mortality rates. RESULTS: In males there were marked increases in all CMM mortality rates between 1975 and 1994, followed by their decrease during 1995-98. In contrast, rates rose in females between 1975 and 1998. There were annual increases of 13% in the risk of dying among males and 33% among females. Relative risks of CMM mortality increased with age in both genders: (i) 21.75 (95% confidence interval, CI 17.32-27.30) in males, and (ii) 28.27 (95% CI 24.99-31.97) in females in the > or = 70-year-old group, when compared with those < 35 years. CONCLUSIONS: CMM mortality rates continue to rise in females while males have experienced a recent reversal of this trend in Spain. Emphasizing the danger of overexposure to ultraviolet radiation may be especially important in females. PMID- 12534604 TI - Prevalence of atopic dermatitis in Japanese adults. AB - BACKGROUND: Adult atopic dermatitis (AD) in Japan has become a significant social problem, with as many as one-third of adult patients with severe AD absenting themselves from work or classes due to aggravation of the disease. Reports of such patients have become increasingly common in recent years. Despite the pressing need for epidemiological studies to clarify the prevalence and distribution of AD and to determine its aetiology, no previous research has been carried out on the prevalence of AD within the adult population in Japan. OBJECTIVES: To clarify the prevalence of adult AD in Japan, using the U.K. Working Party's diagnostic criteria. METHODS: The subjects of this study were mostly government officials or their family members visiting the Medical Center of Health Science, Toranomon Hospital in Tokyo for annual health check-ups in the period from September 1997 to August 1998. Questionnaires completed by 10 762 persons (8076 men and 2686 women) aged 30 years or above were analysed. The questionnaire consisted of 14 questions on allergic disease. The U.K. Working Party's diagnostic criteria were used after translation into Japanese. Three types of prevalence were used as indicators of prevalence: point, 1-year and lifetime prevalence. RESULTS: The point prevalence, 1-year prevalence and lifetime prevalence of AD in Japanese adults were 2.9%, 3.0% and 3.3%, respectively. No significant statistical differences were observed between the sexes or among age groups within each sex. The survey indicated that 88.6% of those who had ever had AD were currently affected by active AD, while 93.4% of those who had had at least one episode of AD in the past had experienced an episode over the previous year. CONCLUSIONS: This study gives the first indication of the prevalence of adult AD among the Japanese, based on the U.K. criteria. Both the internal and external validity of this study are believed to be high; it would be safe to conclude that the 1-year prevalence of AD in Japanese adult populations living in urban areas is 3.0%. PMID- 12534605 TI - Histological changes and involvement of apoptosis after photodynamic therapy for actinic keratoses. AB - BACKGROUND: Photodynamic therapy (PDT), which employs a combination of a tumour localizing photosensitizer and visible light, has been used to treat superficial malignancies in the epidermis. OBJECTIVES: To examine histological changes and the role of apoptosis in lesions of actinic keratosis (AK) after PDT using 5 aminolaevulinic acid (ALA) and excimer dye laser. METHODS: After topical ALA-PDT, biopsy specimens were collected from 18 skin lesions in 15 patients with AK. Paraffin-embedded sections of the skin specimens were stained with haematoxylin and eosin. The detection of apoptosis was performed using a terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labelling (TUNEL) method, antiactivated caspase-3 antibody and anti-Fas antibody. RESULTS: One hour after PDT, cells with eosinophilic cytoplasm and markedly stained nuclei were found, and vacuolation of some tumour cells was noted in the lower layer of the epidermis. An infiltrate of lymphocytes and neutrophils was observed in the upper layer of the dermis. One day after PDT, all layers of the epidermis exhibited slightly degenerative necrosis, with shadow cell formation and chromatin condensation around the nuclear membrane in the lower layer of the epidermis. Necrosis in all layers of the epidermis and lymphocyte infiltration in the dermis were found 3 days after PDT. Tumour cells had disappeared and regenerative thickening of the epidermis was observed 7 days after PDT. TUNEL staining revealed apoptosis-positive cells in the epidermis in 8 of 11 specimens obtained 1 day after PDT. Activated caspase-3 expression was noted in the lower layer of the epidermis in four of these eight TUNEL-positive specimens. CONCLUSIONS: Results suggested that apoptosis is involved in tumour cell death after PDT in patients with AK, and that it occurs within 1 day after PDT. PMID- 12534606 TI - Clinical dose and adverse effects of topical steroids in daily management of atopic dermatitis. AB - BACKGROUND: Topical steroids are used as the first-line therapy for atopic dermatitis. OBJECTIVES: To determine the clinical doses of topical steroids for the daily treatment of atopic dermatitis in clinics and to elucidate their adverse effects. PATIENTS AND METHODS: A multicentre retrospective analysis of a series of 1271 patients (210 infants, 546 children, and 515 adolescents and adults) with atopic dermatitis. RESULTS: Less than 89.5 g, 135 g and 304 g of topical steroid were applied in 90% of the patients in the infant, childhood, and adolescent and adult AD groups, respectively, on the entire body during the 6 month treatment period. The majority of patients were controlled well; however, 7% of infant, 10% of childhood and 19% of adolescent and adult patients remained in a very severe or severe state or experienced exacerbation even though they applied larger amounts of topical steroids. With regard to adverse effects, the incidence of telangiectasia on cheeks tended to increase in patients who had a longer duration of disease and who applied more than 20 g to the face during the 6-month treatment period. The steroid-induced atrophy of the antecubital and popliteal fossae was more frequently observed in males than in females. CONCLUSIONS: Topical steroids are useful for treating atopic dermatitis, but a substantial percentage of patients cannot be satisfactorily treated with topical steroids. For such patients, adjustments of dose and rank of topical steroids and other therapeutic adjuncts are necessary. PMID- 12534607 TI - Efficacy of betamethasone valerate mousse in comparison with standard therapies on scalp psoriasis: an open, multicentre, randomized, controlled, cross-over study on 241 patients. AB - BACKGROUND: The scalp is a common area for plaque psoriasis. Corticosteroid-based lotions are the most widely used therapy in this clinical setting. A new formulation of betamethasone valerate 0.12% in a thermophobic, low-residue foam vehicle (Bettamousse trade mark, Mipharm, Italy; BVM) is available for the treatment of scalp dermatoses. OBJECTIVES: In an open, investigator-blinded, multicentre (28 dermatology clinics), randomized, cross-over study, the efficacy, safety and patient acceptability of BVM in scalp psoriasis were evaluated in comparison with standard therapies (ST, i.e. corticosteroids or vitamin D analogues). ST were chosen by each centre according to its usual therapeutic protocols. METHODS: In total, 241 patients with moderate to severe scalp psoriasis participated in the trial. After a 2-week run-in period, each active treatment (BVM or ST) was applied for 4 weeks, with a wash-out period between the two active treatment phases of at least 4 weeks. Efficacy was evaluated by investigators unaware of treatment sequence analysing a 'target' lesion for erythema, scaling, itching and burning using a five-point grading score. Patient treatment acceptability and assessment of the influence on Psoriasis Disability Index were evaluated using an eight-item modified Finlay-Khan questionnaire at baseline and at the end of each treatment period. Safety was evaluated by recording any adverse event occurring during the study duration. BVM was applied twice daily, and ST were applied once or twice daily, according to the approved scheduled regimens. RESULTS: Analyses were by intention-to-treat. Two hundred and ten patients concluded the study. Fifteen patients withdrew from the study during BVM treatment, and 16 during ST (not significantly different). Both treatments were well tolerated. At baseline, the mean +/- SD clinical global score (the 'Sum' score = erythema + scaling + itching + burning) was 7.6 +/- 2.6. The ST chosen were topical corticosteroids (55% of cases; mainly mometasone and betamethasone dipropionate) or calcipotriol lotion (45% of cases). At the end of active treatments, BVM was significantly superior to ST (P < 0.001) in reducing, as compared with baseline, the mean +/- SD Sum score (1.5 +/- 1.9 with BVM and 3.1 +/- 2.7 with ST). During BVM treatment, 88% (95% confidence interval, CI 82 94%) of patients had a complete or nearly complete resolution of scaling in comparison with 66% (95% CI 58-74%) during ST therapy (P < 0.001). BVM was also considered an easier and more convenient formulation to use in comparison with ST (P < 0.01). CONCLUSIONS: BVM is more effective than lotion-based ST commonly used in the treatment of scalp psoriasis, and has higher patient acceptability. PMID- 12534608 TI - Alternative glucocorticoids for use in cases of adverse reaction to systemic glucocorticoids: a study on 10 patients. AB - BACKGROUND: Reactions to systemically administered corticosteroids are rare, despite their widespread use. OBJECTIVES: To identify alternative glucocorticoids for emergency use in patients with adverse reactions to systemic glucocorticoids. METHODS: Ten patients were identified as having adverse reactions after the use of systemic corticosteroids. Skin prick tests and intradermal tests to hydrocortisone (HC) and methylprednisolone (MP), and intradermal tests to betamethasone and dexamethasone, were performed in all patients, and oral challenge tests to betamethasone (n=10) and deflazacort (n=6). RESULTS: Skin prick tests were negative in all patients, whereas intradermal tests to HC and MP were positive in eight; two patients showed only an isolated cutaneous sensitivity to MP. Intradermal tests to betamethasone and dexamethasone were negative, and oral challenge tests were negative in all patients. CONCLUSIONS: Our results suggest the possibility of an IgE-mediated mechanism for allergic reactions to HC and MP, probably due, at least in part, to a steroid-glyoxal. We suggest that betamethasone and deflazacort could be reserved for emergency use in patients with adverse reactions to other corticosteroids. PMID- 12534609 TI - Valaciclovir as a single dose during prodrome of herpes facialis: a pilot randomized double-blind clinical trial. AB - BACKGROUND: Randomized clinical trials of valaciclovir in recurrent herpes labialis are lacking. OBJECTIVES: To determine whether a single course of valaciclovir, i.e. 500, 1000 or 2000 mg, administered during the prodrome of herpes facialis, could be beneficial. METHODS: Three hundred and forty-five out patients with herpes labialis were screened and randomized for a multicentre, double-blind clinical trial. Ninety-six patients had no recurrence after 6 months of follow-up; 249 patients were finally included in the intent-to-treat (ITT) population. The main outcome measure was the rate of aborted episodes at day 3. The three treatment groups were similar at baseline. RESULTS: There was no statistically significant difference between the groups in rates of aborted lesions at day 3 in the ITT population, in particular between the 500 mg and 2000 mg treatment groups. CONCLUSIONS: Although a placebo group was not included in this pilot study, a single dose of valaciclovir was not considered beneficial in patients with recurrent herpes facialis. PMID- 12534610 TI - Mycophenolate mofetil may serve as a steroid-sparing agent for sarcoidosis. AB - Sarcoidosis is an inflammatory disease with potentially severe mucocutaneous manifestations. Mycophenolate mofetil (MMF) is an immunosuppressive drug extensively used in organ transplantation. Its use has been rapidly expanded into autoimmune and inflammatory diseases. We report the first successful and safe use of MMF in five patients with sarcoidosis. PMID- 12534611 TI - A novel homozygous mutation 371delA in TGM1 leads to a classic lamellar ichthyosis phenotype. AB - Malformation of the cornified cell envelope (CCE) arising from mutations of the transglutaminase (TGase) 1 gene (TGM1) is the cause of some cases of lamellar ichthyosis (LI). However, genotype/phenotype correlation in TGM1 mutations has not yet been fully clarified. We report a typical case of LI caused by a novel mutation in TGM1. The patient, a 33-year-old woman, showed thick, lamellar scales on the entire body surface. Immunofluorescence labelling with anti-TGase 1 antibodies was negative in the patient's epidermis. In situ TGase activity assay detected markedly reduced TGase activity in granular layers of the patient's epidermis. Electron microscopy revealed incomplete thickening of the CCE during keratinization in the epidermis. Sequencing of the entire exons and exon-intron borders of TGM1 revealed that the patient was a homozygote for a novel deletion mutation 371delA in exon 3. This mutation leads to a frameshift resulting in a premature termination codon 43 bp downstream from the mutation site. According to the protein modelling of TGase 1, the truncated protein from this mutated allele loses the entire catalytic core domain of TGase 1. Thus, the present homozygous mutation is expected to cause total loss of TGase 1 activity, resulting in large, dark, lamellar scales on the entire body, the classic phenotype of LI, in this patient. PMID- 12534612 TI - An unusual form of primary systemic amyloidosis: amyloid elastosis: report of a case treated by haematopoietic cell transplantation. AB - Amyloid elastosis is a rare variant of primary systemic amyloidosis characterized by amyloid deposited around elastic fibres. Only two cases, with pseudoxanthoma elasticum-like features and fatal outcome, have been reported. A 56-year-old woman presented with polyneuropathy and a diffuse plane xanthoma-like eruption. Light and electron microscopy studies revealed deposits of amyloid L encasing either normal-looking or short, fragmented elastic fibres in the dermis in a pattern characteristic of amyloid elastosis. The patient had medullary plasmocytosis with lambda light chain restricted expression and underwent autologous stem cell transplantation, which resulted in progressive regression of mucocutaneous signs and stabilization of the polyneuropathy. Our case extends the spectrum of clinical and histopathological presentations of amyloid elastosis. Haematopoietic cell transplantation might improve outcome in patients with multisystem disease. PMID- 12534613 TI - Congenital erythropoietic porphyria associated with myelodysplasia presenting in a 72-year-old man: report of a case and review of the literature. AB - Congenital erythropoietic porphyria (CEP) is a rare autosomal recessive disease owing to the deficient activity of uroporphyrinogen III synthase, the fourth enzyme in the porphyrin-haem synthetic pathway. Of the porphyrias, it is the most mutilating type, usually presenting early in life. To date, 12 documented cases of adult onset CEP have been reported. We report the second oldest documented patient with late onset CEP with incidental findings of thrombocytopenia and myelodysplasia with bone-marrow sideroblasts. We further discuss several current and future treatment options for this therapeutically challenging disease. PMID- 12534614 TI - Localized lichen myxoedematosus (papular mucinosis) associated with morbid obesity: report of two cases. AB - Cutaneous diseases are often found in obese patients but, to our knowledge, mucinous disorders have not been previously reported in association with obesity. Two cases of localized lichen myxoedematosus (papular mucinosis) in two women with morbid obesity are described. Both patients underwent a low-calorie diet for a 1-year period in one case, and for 4 months in the other one, as the only treatment. There was complete resolution of cutaneous lesions at the same time that an important weight loss was observed. Nevertheless, although spontaneous regression is not frequent, it could not be disregarded in either of these two cases. PMID- 12534615 TI - Why do I have psoriasis? PMID- 12534616 TI - The Woronoff ring in psoriasis. PMID- 12534617 TI - Irritant dermatitis due to prolonged contact with Oilatum Plus. PMID- 12534618 TI - Severe urticaria due to recombinant interferon beta-1a. PMID- 12534619 TI - Allergic contact dermatitis to broccoli. PMID- 12534620 TI - Bilateral nail dystrophy associated with vibrating tool use. PMID- 12534621 TI - Mycetoma caused by Scytalidium dimidiatum. PMID- 12534622 TI - An audit of the use of self-administered adrenaline syringes in the east end of Glasgow. PMID- 12534623 TI - Bupropion and generalized acute urticaria: eight cases. PMID- 12534624 TI - Erythromelalgia in a patient with idiopathic thrombocytopenic purpura. PMID- 12534625 TI - Shiitake dermatitis. PMID- 12534626 TI - Malignant proliferating trichilemmal tumour associated with human papillomavirus type 21 in epidermodysplasia verruciformis. PMID- 12534627 TI - Detection of immediate-type reaction to the epitope of beta-lactam antibiotics by the quick patch test. PMID- 12534628 TI - Merkel cell carcinoma composed of small, intermediate and squamous cell foci showing mutually exclusive expression of neuroendocrine markers and cytokeratin 20. PMID- 12534629 TI - Multiple dermatofibroma-like lesions in a human immunodeficiency virus-positive patient coinfected with visceral leishmaniasis. PMID- 12534630 TI - Pigmented wart showing fibrillar intracytoplasmic inclusion bodies. PMID- 12534631 TI - Localization of human beta-defensin-2 and human neutrophil peptides in superficial folliculitis. PMID- 12534634 TI - Good clinical practice: a nuisance, a help or a necessity for clinical pharmacology? AB - This article reviews the impact of good clinical practice (GCP) on clinical pharmacology with particular reference to the new European Union Clinical Trial Directive. The Directive will be applied to both commercial and noncommercial studies on medicinal products for human use. The Directive requires that GCP should be used in all clinical trials except noninterventional studies. GCP is likely to follow the International Conference on Harmonization GCP guidelines in many aspects. GCP will enforce tighter guidelines on ethical aspects of a clinical study. Higher standards will be required in terms of comprehensive documentation for the clinical protocol, record keeping, training, and facilities including computers. Quality assurance and inspections will ensure that these standards are achieved. The additional requirements of GCP are discussed and any advantage to the study subject. The impact of the new Directive within the Research Governance Framework of the UK Department of Health is reviewed. PMID- 12534635 TI - Pharmacokinetics of oral betaine in healthy subjects and patients with homocystinuria. AB - AIMS: Large oral doses of betaine have proved effective in lowering plasma homocysteine in severe hyperhomocysteinaemia. The pharmacokinetic characteristics and metabolism of betaine in humans have not been assessed and drug monitoring for betaine therapy is not available. We studied the pharmacokinetics of betaine and its metabolite dimethylglycine (DMG) in healthy subjects and in three patients with homocystinuria. METHODS: Twelve male volunteers underwent an open label study. After one single administration of 50 mg betaine kg-1 body weight and during continuous intake of twice daily 50 mg kg-1 body weight, serial blood samples and 24 h urines were collected to determine betaine and DMG plasma concentrations and urinary excretion, respectively. Patients were evaluated after one single dose of betaine. RESULTS: We found rapid absorption (t(1/2),abs 00.28 h, s.d. 0.17) and distribution (t(1/2), lambda1 00.59 h, s.d. 0.22) of betaine. A Cmax of 0.94 mmol l-1 (s.d. 0.19) was reached after tmax 00.90 h (s.d. 0.33). The elimination half life t(1/2), z was 14.38 h (s.d. 7.17). After repeated dosage, t(1/2), lambda1 (01.77 h, s.d. 0.75) and t(1/2), z (41.17 h, s.d. 13.50) increased significantly (95% CI 0.73, 01.64 h and 19.90, 33.70 h, respectively), whereas absorption remained unchanged. DMG concentrations increased significantly after betaine administration and accumulation occurred to the same extent as with betaine. Renal clearance was low and urinary excretion of betaine was equivalent to 4% of the ingested dose. Distribution and elimination kinetics in homocystinuric patients appeared to be accelerated. CONCLUSIONS: Betaine plasma concentrations change rapidly after ingestion. Elimination half-life increased during continuous dosing over 5 days. Betaine is mainly eliminated by metabolism. More pharmacokinetic and pharmacodynamic studies in hyperhomocysteinaemic patients are needed to refine the current treatment with betaine. PMID- 12534636 TI - Rectal ropivacaine is absorbed proportionally to the dose, with low intraindividual variability. AB - AIMS: This study investigated the absorption characteristics and the tolerability of rectally administered ropivacaine, a local anaesthetic, intended as a new local therapy for ulcerative colitis. METHODS: Thirty-two healthy volunteers participated in a randomized, placebo-controlled study. In study phase 1 (n = 16, double-blind, crossover) single rectal doses of ropivacaine corresponding to 50, 100 and 200 mg base were given as 20-ml gel enemas. Eight of these subjects also received an i.v. infusion corresponding to 20 mg (2H3)ropivacaine base given with the last rectal dose. In study phase 2 (n = 16, single-blind, crossover) the same rectal doses were given but formulated in 20, 40 and 80 ml gel, respectively. Peripheral venous plasma samples and urine were collected over 12 h after dosing and analysed for ropivacaine base by gas chromatography and (2H3)ropivacaine by gas chromatography-mass spectrometry. Ropivacaine and metabolites were analysed in urine by reversed phase liquid chromatography. RESULTS: AUC was proportional to the dose with a point estimate [95% confidence interval (CI)] for the increase, after doubling the dose, of 1.91 (1.66-2.20) and 1.95 (1.78-2.13) in study phases 1 and 2, respectively. The increase in Cmax was also proportional to the dose with corresponding results of 1.76 (1.52-2.04) and 1.84 (1.70-1.99). The volume of the rectal formulation had no influence on either the extent or the time course of absorption. The mean (s.d.) absolute bioavailability (%F) was 56 (18)%. AUC and Cmax showed a two- to three-fold lower intra- than interindividual variability. Zero-order kinetics dominated the first 4 h of the absorption phase. Thereafter first-order kinetics were observed. The terminal half-lives were similar between the rectal doses and were longer than that after the i.v. administration, indicating an absorption-dependent half-life. The main urinary metabolite was 3-hydroxyropivacaine corresponding to about 23% of the dose. The subjects had no difficulties in retaining the doses and rectal administration of ropivacaine was well tolerated, both locally and systemically. CONCLUSIONS: Plasma drug concentrations were proportional to the dose after rectally administered doses corresponding to 50-200 mg ropivacaine base in a gel formulation. The drug was well-tolerated. Mean bioavailability was about 60% and not influenced by variations in the enema volume. Initial absorption seemed to follow zero-order kinetics and then first-order kinetics after about 4 h. Cmax and AUC showed considerably less intra- compared with inter-individual variability, resulting in more consistent plasma concentrations within subjects. PMID- 12534637 TI - Population pharmacokinetics of theophylline during paediatric extracorporeal membrane oxygenation. AB - AIMS: To determine the population pharmacokinetics of theophylline during extracorporeal membrane oxygenation (ECMO) from routine monitoring data. METHODS: Retrospective data were collected from 75 term neonates and children (age range 2 days to 17 years) receiving continuous infusions of aminophylline (mean rate 9.2 +/- 2.6 micro g kg-1 min-1) during ECMO. A total of 160 plasma concentrations (range 1-8 per patient), sampled at time intervals ranging from 10 h to 432 h, were included. Population PK analysis and model building were carried out using WinNonMix Professional (Version 2.0.1). Cross-validation was used to evaluate the validity and predictive accuracy of the model. RESULTS: A one-compartment model with first order elimination combined with an additive error model was found to best describe the data. Of the covariables tested, bodyweight significantly influenced clearance and volume of distribution, whereas age was an important determinant of clearance, as adjudged by the differences in the -2 x log likelihood (P < 0.005) and the residual error value. The final model parameters were estimated as: clearance (l h-1) = 0.023 x bodyweight (kg) + 0.000057 x age (days) and volume of distribution (l) = 0.57 x bodyweight (kg). The interindividual variability in clearance and volume of distribution was 38% and 40%, respectively. The residual error corresponded to a standard deviation of 3.6 mg l-1. Cross-validation revealed a median (95% confidence interval) model bias of 9.4% (2.9, 16.5%) and precision of 29.5% (24.8, 36.0%). CONCLUSIONS: The estimated clearance is significantly lower, and volume of distribution higher, than previously reported in non-ECMO patients of similar age. These differences are probably a result of the expanded circulating volume during ECMO and altered renal and hepatic physiology in this critically ill group. Large interindividual variability reflects the heterogeneous nature of patients treated on ECMO. PMID- 12534638 TI - Population pharmacokinetic analysis of meloxicam in rheumatoid arthritis patients. AB - AIM: To perform a nonlinear mixed effect modelling (NONMEM) population pharmacokinetic analysis of meloxicam plasma concentrations in rheumatoid arthritis (RA) patients participating in three clinical trials, and to evaluate the effects of age, weight, gender and concomitant medications on meloxicam pharmacokinetics. METHODS: Meloxicam was administered to RA patients once daily for 3 weeks or 6 months at doses between 7.5 and 60 mg. Plasma samples were obtained at least 7 days after the first dose and meloxicam plasma concentrations were quantified by h.p.l.c. RESULTS: NONMEM analysis was conducted on plasma samples derived from 586 patients. A one-compartmental model was found to describe the data adequately. For a typical subject in the population, a clearance of 0.377 l h-1 (95% confidence interval (CI) 0.0304-0.449) in males and 0.347 l h-1 (95% CI 0.274-0.419) in females was obtained. The volume of distribution was estimated to be 14.9 l. The findings were corroborated by subsequent analysis using WinBUGS. Analysis of covariates showed that age and gender both significantly (P < 0.005) affected clearance. The effect of age was relatively small and a dose adjustment of <10% was deemed unnecessary. Differences between males and females were attributed to differences in weight. No clinically relevant drug-drug interactions were found, although sulphasalazine and glucocorticoids both significantly (P < 0.005) affected meloxicam clearance (+19% and - 12%, respectively). The mechanisms by which these agents affect meloxicam clearance remain to be elucidated. CONCLUSIONS: The population pharmacokinetic meloxicam data from patients with RA gave similar results to those obtained from phase I trials. However, uncommon drug interactions may not be detected in phase I trials because of the small number of observations made. PMID- 12534639 TI - Biomarkers for the effects of benzodiazepines in healthy volunteers. AB - Studies of novel centrally acting drugs in healthy volunteers are traditionally concerned with kinetics and tolerability, but useful information may also be obtained from biomarkers of clinical endpoints. A useful biomarker should meet the following requirements: a consistent response across studies and drugs; a clear response of the biomarker to a therapeutic dose; a dose-response relationship; a plausible relationship between biomarker, pharmacology and pathogenesis. In the current review, all individual tests found in studies of benzodiazepine agonists registered for anxiety in healthy volunteers since 1966 were progressively evaluated for compliance with these requirements. A MedLine search yielded 56 different studies, investigating the effects of 16 different benzodiazepines on 73 different (variants of ) neuropsychological tests, which could be clustered into seven neuropsychological domains. Subjective and objective measures of alertness were most sensitive to benzodiazepines. The most consistent effects were observed on saccadic peak velocity (SPV) and visual analogue scores ( VAS) of alertness, where 100% and 79% of all studies respectively showed statistically significant effects. A dose-response relationship could be constructed for temazepam and SPV, which was used to determine dose equivalencies relative to temazepam, for seven different benzodiazepines. These dose equivalencies correlated with the lowest recommended daily maintenance dose (r2 = 0.737, P < 0.05). This relationship between SPV reduction and clinical efficacy could reflect the clinical practice of aiming for maximum tolerated levels, or it could represent a common basis behind SPV reduction and anxiolytic activity for benzodiazepines (probably sedation). The number of tests used in human psychopharmacology appears to be excessive and their sensitivity and reproducibility low. PMID- 12534640 TI - Pharmacokinetics of diclofenac and inhibition of cyclooxygenases 1 and 2: no relationship to the CYP2C9 genetic polymorphism in humans. AB - AIMS: The cytochrome P450 enzyme CYP2C9 catalyses the 4'-hydroxylation of the nonsteroidal analgesic drug diclofenac in humans. We studied the influences of the known amino acid variants, CYP2C9*2 (Arg144Cys) and CYP2C9*3 (Ile359Leu), on diclofenac pharmacokinetics after a 50-mg oral dose of diclofenac in healthy volunteers. As a surrogate marker of diclofenac activity, the ex vivo formation of prostaglandin E2 and thromboxane B2, which reflects COX-2 and COX-1 activity, was measured. METHODS: Genotyping was performed in 516 healthy volunteers to obtain 20 participants with all allelic combinations of the two CYP2C9 variants Arg144Cys (*2) and Ile359Leu (*3). Diclofenac and 4'-hydroxydiclofenac were quantified in plasma by reversed phase h.p.l.c. after oral intake of 50 mg diclofenac. Concentrations of thromboxane B2 (TxB2) and prostaglandin E2 (PGE2) were measured by immunoassays. RESULTS: There was no evidence of impaired metabolism of oral diclofenac in heterozygous and homozygous carriers of the CYP2C9 alleles *2 and *3 compared with the wild type (mean CL/F (95% CI) 20.5 (11, 30) l h-1 for *1/*1, 29.9 (19, 40) l h-1 for *1/*2, 30.0 (4, 56) l h-1 for *2/*2, 22.6 (12, 33) l h-1 for *1/*3, 23.5 (11, 37) l h-1 for *3/*3 and 37.3 ( 15, 89) l h-1 in *2/*3). Furthermore, plasma concentrations of the metabolite 4' hydroxydiclofenac were not lower in carriers of the CYP2C9 low-activity alleles *2 and *3 compared with carriers of the CYP2C9*1/*1 genotype. Marked diclofenac mediated inhibition of COX-1- and COX-2 activity was detected in all individuals independent of CYP2C9 genotype. CONCLUSIONS: Polymorphisms of the CYP2C9 gene had no discernible effect on the pharmacokinetics and pharmacodynamics of diclofenac. The question of whether enzymes other than CYP2C9 play a major role in diclofenac 4'-hydroxylation in vivo or whether 4'-hydroxylation is not a rate-limiting step in diclofenac elimination in vivo, or whether the effect of the CYP2C9 polymorphisms is substrate-dependent, needs further investigation. PMID- 12534641 TI - NAT2 and CYP1A2 phenotyping with caffeine: head-to-head comparison of AFMU vs. AAMU in the urine metabolite ratios. AB - AIMS: (i) To compare the phenotyping of healthy subjects for NAT2 and CYP1A2 activities with caffeine, by the simultaneous assay of the urinary metabolites AFMU and AAMU, and (ii) to ascertain whether NAT2 and CYP1A2 phenotyping is influenced by the use of AFMU or AAMU in the metabolite ratio. METHODS: Thirty five healthy subjects (16 men, 19 women) participated to the study. Caffeine metabolite concentrations were measured in urine collected 8 h after 2.5 mg kg-1 caffeine intake using a new validated h.p.l.c. method. The metabolite ratios AFMU/1X, AFMU/(AFMU+1X+1U), AAMU/1X, AAMU/(AAMU+1X+1 U), and (AFMU+1U+1X)/17U, (AAMU+1U+1X)/17U were determined as indices of NAT2 and CYP1A2 activity, respectively. RESULTS: Slow and rapid acetylators were similarly identified using the four NAT2 metabolite ratios in 139 out of 140 measurements. An appreciable amount of AAMU was present in urine that was immediately acidified and analysed. Consequently, the ratio using AFMU was lower than that using total AAMU following transformation of AFMU in basic conditions. The proportion of AFMU in urine analysed immediately expressed as AFMU/(AFMU+AAMU) ratio did not correlate with urine pH, but was a function of the acetylation phenotype, with a low intergroup variability (64 +/- 3% and 32 +/- 5%, for rapid and slow acetylators, respectively; P < 0.00001, anova). Regarding CYP1A2 activity, a good correlation (r = 0.99) was observed between the metabolite ratios calculated from AFMU and AAMU, although the ratios calculated from AFMU were proportionately and systematically lower P < 0.00001, paired t-test, slope 1.2). CONCLUSIONS: This study demonstrates that both AFMU and AAMU can be used for NAT2 and CYP1A2 metabolite ratio determinations. The reported conversion of AFMU into AAMU is unlikely to explain the large amount of AAMU in urine that was acidified and analysed immediately after voiding. The results suggest that AAMU is formed not solely through a nonenzymatic hydrolysis in urine, but in vivo by a NAT2 phenotype-dependent pathway. PMID- 12534642 TI - Polymorphisms in the cytochrome P450 CYP1A2 gene (CYP1A2) in colorectal cancer patients and controls: allele frequencies, linkage disequilibrium and influence on caffeine metabolism. AB - AIMS: Several single nucleotide polymorphisms (SNPs) of the cytochrome P450 enzyme 1A2 gene (CYP1A2) have been reported. Here, frequencies, linkage disequilibrium and phenotypic consequences of six SNPs are described. METHODS: From genomic DNA, 114 British Caucasians (49 colorectal cancer cases and 65 controls) were genotyped for the CYP1A2 polymorphisms -3858G-->A (allele CYP1A2*1C), -2464T-->delT (CYP1A2*1D), -740T-->G (CYP1A2*1E and *1G), -164A-->C (CYP1A2*1F), 63C-->G (CYP1A2*2), and 1545T-->C (alleles CYP1A2*1B, *1G, *1H and *3), using polymerase chain reaction-restriction fragment length polymorphism assays. All patients and controls were phenotyped for CYP1A2 by h.p.l.c. analysis of urinary caffeine metabolites. RESULTS: In 114 samples, the most frequent CYP1A2 SNPs were 1545T-->C (38.2% of tested chromosomes), -164A-->C (CYP1A2*1F, 33.3%) and -2464T-->delT (CYP1A2*1D, 4.82%). The SNPs were in linkage disequilibrium: the most frequent constellations were found to be -3858G/-2464T/ 740T/-164A/63C/1545T (61.8%), -3858G/-2464T/-740T/-164C/63C/1545C (33.3%), and 3858G/-2464delT/-740T/-164A/63C/1545C (3.51%), with no significant frequency differences between cases and controls. In the phenotype analysis, lower caffeine metabolic ratios were detected in cases than in controls. This was significant in smokers (n = 14, P = 0.020), and in a subgroup of 15 matched case-control pairs (P = 0.007), but it was not significant in nonsmokers (n = 100, P = 0.39). There was no detectable association between CYP1A2 genotype and caffeine phenotype. CONCLUSIONS: (i) CYP1A2 polymorphisms are in linkage disequilibrium. Therefore, only -164A-->C (CYP1A2*1F) and -2464T-->delT (CYP1A2*1D) need to be analysed in the routine assessment of CYP1A2 genotype; (ii) in vivo CYP1A2 activity is lower in colorectal cancer patients than in controls, and (iii) CYP1A2 genotype had no effect on phenotype (based on the caffeine metabolite ratio). However, this remains to be confirmed in a larger study. PMID- 12534643 TI - Diabetes mellitus increases the in vivo activity of cytochrome P450 2E1 in humans. AB - AIMS: Cytochrome P450 2E1 (CYP2E1) is thought to activate a number of protoxins, and has been implicated in the development of liver disease. Increased hepatic expression of CYP2E1 occurs in rat models of diabetes but it is unclear whether human diabetics display a similar up-regulation. This study was designed to test the hypothesis that human diabetics experience enhanced CYP2E1 expression. METHODS: The pharmacokinetics of a single dose of chlorzoxazone (500 mg), used as an index of hepatic CYP2E1 activity, was determined in healthy subjects (n = 10), volunteers with Type I (n = 13), and Type II (n = 8) diabetes mellitus. Chlorzoxazone and 6-hydroxychlorzoxazone in serum and urine were analysed by high performance liquid chromatography. The expression of CYP2E1 mRNA in peripheral blood mononuclear cells was quantified by reverse transcriptase-polymerase chain reaction. RESULTS: The mean +/- s.d. (90% confidence interval of the difference) chlorzoxazone area under the plasma concentration-time curve was significantly (P 85%) of the total inhibitors. Ketoconazole did not affect the proportion of circulating active or total inhibitors accounted for by circulating rosuvastatin. CONCLUSIONS: Ketoconazole did not produce any change in rosuvastatin pharmacokinetics in healthy subjects. The data suggest that neither cytochrome P450 3A4 nor P-gp mediated transport contributes to the elimination of rosuvastatin. PMID- 12534646 TI - Pharmacokinetic interaction between amprenavir and delavirdine after multiple dose administration in healthy volunteers. AB - AIMS: To evaluate the safety and the pharmacokinetic interaction between amprenavir and delavirdine after multiple dose administration in healthy volunteers. METHODS: This was a prospective, open-label, randomized, controlled, two-sequence, two-period multiple dose study with 18 healthy subjects. Volunteers were randomly assigned to amprenavir, 600 mg twice a day, or delavirdine, 600 mg twice a day, for 10 days, followed by both drugs for another 10 days with pharmacokinetic evaluation on day 10 and day 20. Adverse events were recorded throughout the study. RESULTS: Amprenavir decreased all the delavirdine pharmacokinetic parameters apart from tmax. Delavirdine C12h dropped from 7,916 to 933 ng ml-1 (median decrease 5,930 ng ml-1, 95% CI 3,013, 8,955 ng ml-1). A decrease in amprenavir t(1/2) was also seen leading to almost identical median amprenavir C24h values. No serious clinical adverse events were observed during the study. The most frequently reported effects were gastrointestinal symptoms, headache, fatigue and rash. CONCLUSIONS: Amprenavir is an effective inducer of delavirdine metabolism, probably through its effect on hepatic CYP3A4. This could have consequences in other drug-drug interaction situations. Delavirdine is an inhibitor of amprenavir metabolism. The regimen of amprenavir 600 mg and delavirdine 600 mg twice a day is not recommended when an antiretroviral effect from delavirdine is required. PMID- 12534647 TI - MMR vaccine and idiopathic thrombocytopaenic purpura. AB - AIMS: To estimate the relationship between idiopathic thrombocytopaenic purpura (ITP) and the measles, mumps and rubella (MMR) vaccination in children; calculating the relative risk estimate for ITP with in 6 weeks after MMR vaccination and the attributable risk of ITP within 6 weeks after MMR vaccination. METHODS: Using the General Practice Research Database we identified children with a first-time diagnosis of ITP from a base population of children aged less than 6 years between January 1988 and December 1999. After describing the characteristics of all the children identified with ITP, we focused on cases aged 13-24 months to perform a population-based, case-control analysis to estimate the relative risk of developing ITP within 6 weeks after MMR vaccination. We also calculated the risk of ITP attributable to the MMR vaccination. RESULTS: Sixty-three children with a first time diagnosis of ITP were identified; 23 cases were between 13 and 24 months old. The relative risk estimate for ITP within 6 weeks after MMR vaccination, compared to the combined group of unvaccinated children and children vaccinated with MMR more than 26 weeks previously was 6.3 (95% CI 1.3-30.1). The attributable risk of developing ITP within 6 weeks after MMR vaccination was estimated to be 1 in 25,000 vaccinations (95% confidence interval 21,300, 89,400). CONCLUSION: This study confirms the increased risk of ITP within 6 weeks after MMR vaccination. However, the attributable risk of ITP within 6 weeks after MMR vaccination is low. PMID- 12534648 TI - Unwanted pregnancy on self-medication with St John's wort despite hormonal contraception. PMID- 12534649 TI - No turning back: the College, health care and politics. PMID- 12534650 TI - Is it time to introduce repetitive transcranial magnetic stimulation into standard clinical practice for the treatment of depressive disorders? AB - OBJECTIVE: To examine issues relating to the potential introduction of repetitive transcranial magnetic stimulation (rTMS) into clinical practice as a treatment for depression. METHOD: A review of the outcomes literature accompanied by an analysis of issues relating to the potential advantages and pitfalls of the introduction of rTMS as a treatment strategy. RESULTS: Evidence is progressively accumulating that rTMS has antidepressant properties that are clinically relevant. These effects are biologically plausible and supported by basic research. Patients with therapy-resistant depression have few treatment alternatives and experience significant suffering, thus justifying the early introduction of a new treatment such as rTMS for this patient group. However, this must be balanced by a need to foster considerable further research and not to raise expectations unreasonably. CONCLUSIONS: It is timely for rTMS to be made more available to patients with treatment-resistant mood disorders. This need not be limited to clinical research trials but should only occur in medical settings where continual evaluation and research is conducted. PMID- 12534652 TI - Allocation of resources and psychosis. AB - OBJECTIVE: This commentary reviews current expenditure on psychosis in Australia, identifies discretionary expenditure that could be used more efficiently, discusses the factors influencing resource allocation and intervention selection decisions, and suggests priorities for change. METHOD: Cost-of-illness findings from the Low Prevalence Disorders Study (LPDS), and related service use and psychosocial data, are used to highlight patterns of expenditure on psychosis and potential resource allocation issues. Arguments are also presented suggesting that mental health resource allocation in Australia should be informed primarily by treatment efficiency, equity and humanitarian considerations, not differences in the global burden of disease. However, our evidence-base about the effectiveness and costs associated with individual treatments, programmes, and organizational structures is also shown to be limited. CONCLUSIONS: The patterns of service use and expenditure on psychosis suggest certain imbalances, including an over-reliance on hospitalization, low levels of supported community accommodation, and inadequate provision of evidence-based psychosocial treatments, rehabilitation and supported employment programmes. We need to identify and develop efficient interventions and programmes, re-orientate our services to better utilize those interventions, increase community awareness, improve monitoring of outcomes and costs, and undertake timely evaluations at multiple levels, from the individual to the societal perspective. PMID- 12534653 TI - Motherhood and schizophrenic illnesses: a review of the literature. AB - OBJECTIVE: To provide an overview of the current knowledge on the impact of motherhood on women with schizophrenia and schizoaffective disorder. METHOD: The published literature was selectively reviewed and assessed, based on a complete MEDLINE and PsychLIT (1971 to current) search, including English and non-English journals and books. RESULTS: Research to date into motherhood and schizophrenic illnesses has been limited by a number of methodological constraints, limiting the ability to draw conclusions and the prevention of relapses and mother-infant difficulties. These constraints have included: a paucity of prospective studies with initial, antenatal recruitment; variable definitions of the length of the puerperium; significant changes in psychiatric classification; the heterogeneity of postpartum psychotic disorders, with the majority being mood or schizoaffective disorder rather than schizophrenia; selection biases inherent in studying mother-baby unit inpatients; difficulties in life events research in general, such as its retrospective nature and confounding, illness factors; and the specificity versus non-specificity of childbirth as a unique or discrete life event. CONCLUSIONS: Further study is required to explore: the impact of child care, parenting and having a partner on the course of women with schizophrenic and schizoaffective disorders during the first postpartum year; whether women with postpartum relapses of these mental illnesses are likely to have slower recoveries than those women with the same diagnoses but without young children; and protective factors against postpartum relapse. PMID- 12534654 TI - Costs of schizophrenia and other psychoses in urban Australia: findings from the Low Prevalence (Psychotic) Disorders Study. AB - OBJECTIVE: To estimate the costs associated with the treatment and care of persons with psychosis in Australia based on data from the Low Prevalence Disorders Study (LPDS), and to identify areas where there is potential for more efficient use of existing health care resources. METHOD: The LPDS was a one-month census-based survey of people with psychotic disorders in contact with mental health services, which was conducted in four metropolitan regions in 1997-1998. Mental health and service utilization data from 980 interviews were used to estimate the economic costs associated with psychotic disorders. A prevalence based, 'bottom-up' approach was adopted to calculate the government and societal costs associated with psychosis, including treatment and non-treatment related costs. RESULTS: Annual societal costs for the average patient with psychosis are of the order of 46,200 Australian dollars , comprising 27,500 Australian dollars in lost productivity, 13,800 Australian dollars in inpatient mental health care costs and 4900 Australian dollars in other mental health and community services costs. Psychosis costs the Australian government at least 1.45 billion Australian dollars per annum, while societal costs are at least 2.25 billion Australian dollars per annum (including 1.44 billion Australian dollars for schizophrenia). We also report relationships between societal costs and demographic factors, diagnosis, disability and participation in employment. CONCLUSIONS: Current expenditure on psychosis in Australia is probably inefficient. There may be substantial opportunity costs in not delivering effective treatments in sufficient volume to people with psychotic disorders, not intervening early, and not improving access to rehabilitation and supported accommodation. PMID- 12534655 TI - Schema-focused cognitive therapy for bipolar disorder: reducing vulnerability to relapse through attitudinal change. AB - OBJECTIVE: Acceptance of, and adaptability to illness, are major determinants of adherence to treatment and functional recovery. This paper addresses the major psychosocial factors associated with bipolar disorder and the role of psychological interventions in symptom management and adaptability to the illness experience. A new model is presented highlighting the role of developmental experiences and temperament in determining reactions to bipolar disorder. The authors propose that by addressing reactions to the illness experiences and effects on self-concept through schema-focused cognitive therapy, functional recovery is more likely to occur among those patients functioning below expectation. METHOD: A systematic review of the current literature including an Index Medicus/MEDLINE search was conducted, focusing on risk factors, cognitive vulnerabilities and triggers associated with bipolar disorder. Psychological treatments available for the treatment of bipolar disorder are reviewed and details of a novel schema-focused cognitive model for this condition are presented. Traditional models of adaptation to chronic illness are outlined and incorporated into the proposed model. Schema-focused cognitive therapy is proposed as an approach to help patients reduce cognitive vulnerability to relapse in addition to adopting effective mood management strategies. RESULTS AND CONCLUSIONS: There is a need for psychological treatments which reduce the risks associated with poor functionality in patients with bipolar disorder. Schema focused cognitive therapy specifically targets the temperament, developmental experiences and cognitive vulnerabilities that determine adjustment to illness. This proposed treatment, combined with pharmacotherapy, may offer new psychotherapeutic options for the future. PMID- 12534656 TI - The subjective experience of patients who received electroconvulsive therapy. AB - OBJECTIVE: Despite the vast amount of scientific literature available on electroconvulsive therapy (ECT), there is little qualitative focus upon the patients' subjective experience of this procedure. Using an exploratory descriptive methodology, this study aims to provide a more unique insight into what certain patients actually think of ECT. METHOD: Semistructured interviews were conducted to explore eight patients' opinions and experiences of ECT. Interviews were subjected to analysis by a five-step framework approach that identified prominent themes in relation to five broad questions and in conjunction with issues raised by the subjects themselves. RESULTS: Eleven major themes were identified. Four of these were chosen for discussion, not only as the most prevalent themes (in terms of how frequently they were mentioned by the subjects), but also as the most striking (in regards to the intensity of emotions evoked, or their influence on their perception of ECT as a future treatment option). The four themes are fear of ECT, attribution of cognitive decline and memory loss to ECT, positive ECT experiences, and patients' suggestions. CONCLUSIONS: Using such a qualitative approach, the depth of the information obtained has revealed new perspectives on how patients perceive the experience of ECT. Fears reported by patients present an opportunity to address specific areas of the procedure that generate the most angst. These were closely associated with recommendations that many patients proposed throughout the interviews. Patients' perceptions of the cognitive effects of ECT do not necessarily correspond with those commonly reported in the literature on ECT. Positive experiences with ECT were more complex than simply its efficacy. There is a need for future research in order to explore and address patients' experiences of ECT. PMID- 12534657 TI - Use of antipsychotic medications in Australia between July 1995 and December 2001. AB - OBJECTIVE: To examine trends in the use of oral conventional, depot and atypical anti-psychotic medications in Australia between July 1995 and December 2001. METHOD: For each six-month interval between July 1995 and December 2001, prescription data obtained from the Health Insurance Commission of Australia were converted into a measure of drug utilization expressed as the number of defined daily doses per thousand population per day (DDDs/1000/day). RESULTS: Between July 1995 and December 2001, use of atypical medications increased from an estimated 0.27 to an estimated 3.83 DDDs/1000/day. Use of oral conventional medications decreased from 1.76 to 0.91 DDDs/1000/day, while use of depot medications decreased from 1.38 to 0.96 DDDs/1000/day. During the latter half of 2001, atypical medications accounted for 67.3%, oral conventional medications for 16.0%, and depot medications for 16.7% of total prescriptions for antipsychotic medications dispensed through community pharmacies in Australia. The increased use of atypical medications was accounted for largely by a dramatic increase in the use of olanzapine. Between July 2001 and December 2001, prescriptions for olanzapine accounted for 65.0% of total prescriptions for atypical antipsychotic medications. CONCLUSIONS: The results of the present study confirm that atypical antipsychotic medications have replaced conventional medications as the first line pharmacological treatment for psychotic illness in Australia. While this change in prescribing practice is gratifying, the overwhelming preference for the use of olanzapine among clinicians prescribing antipsychotic medication is surprising and is of some concern given the significant increases in body weight that can accompany long-term use of this medication. PMID- 12534658 TI - Vascular risk to late-life depression: evidence from a longitudinal community study. AB - OBJECTIVE: To determine prospectively relationships between minor cerebrovascular episodes and depressive symptoms in a community cohort of older persons. METHOD: In 1988-1989, baseline measurements of vascular risk factors and depressive symptoms were obtained in older community residents (mean age = 67). At 10-year follow-up, three subgroups of subjects still residing in the community were re assessed: those who had suffered a transient ischaemic attack (TIA) (n = 16) in the intervening period; those with hypertension but no TIAs (n = 38); and, those with neither TIAs nor hypertension (n = 40). RESULTS: Of the 16 persons with depressive symptoms at 10-year follow-up, only three had reported depressive symptoms initially. Subjects who had experienced TIAs during the longitudinal phase had higher rates of depressive symptoms than the subjects from the other two groups (38%vs 13%, p < 0.05). CONCLUSIONS: This study supports the notion that cerebrovascular incidents predispose to late-onset depression in older persons residing in the community. Intrinsically, this provides epidemiological support for the validity of the concept of 'vascular depression'. PMID- 12534659 TI - Age-at-first-registration and heterogeneity in affective psychoses. AB - BACKGROUND: Previous research into age of onset in affective disorders has produced conflicting results. This paper examines the influence of heterogeneity on the age-at-first-registration distribution for the ICD-9 diagnostic group 'affective psychosis'. METHOD: For 1979-1991, data for age-at-first-registration for 4985 individuals diagnosed with affective psychosis (ICD-9 296.x) were extracted from a name-linked mental health register. These data were divided into (i) '296.1 only', a category used to code unipolar depression (males = 700; females = 1321); and (ii) '296 other', all 296 cases other than 296.1 (males = 1280; females = 1684). Inception rates for each 5-year age division were adjusted for the background population age-structure as a rate per 100,000 population. RESULTS: The age-at-first-registration distribution for affective psychosis has a wide age range, with women outnumbering men. There is a near-linear increase in inception rates for both men and women with 296.1 only, while the bulk of those with affective psychoses (296 other) have an inverted U-shaped age distribution. Males have an earlier modal age-at-first-registration for 296 other compared to females. CONCLUSION: The heterogeneity in terms of subtypes and sex in affective psychosis clouds the interpretation of age-at-first-registration. Separating those with unipolar psychotic depression from other subclassifications and differentiating by sex may provide clues to factors that precipitate the onset of affective psychosis. PMID- 12534660 TI - Depression in people living with HIV/AIDS attending primary care and outpatient clinics. AB - OBJECTIVE: Our aim was to gain an estimate of the rate of depressive disorder in patients with HIV/AIDS attending general practice and to investigate factors associated with depression. A further objective was to determine the ability of non-mental health medical practitioners to detect depressive symptoms in their patients with HIV/AIDS. METHODS Participants comprised 322 persons living with HIV/AIDS ( (PLWHA); 13 females, 309 males; mean age 41.4, SD = 8.9) who were recruited from four general practice clinics specializing in HIV medicine and from an infectious diseases clinic. Medical, psychiatric and sociodemographic data were obtained. In addition, participants completed the Inventory to Diagnose Depression (IDD), a self-report measure to detect depression. RESULTS: Twenty-two per cent of the sample met criteria for a current Major Depressive Episode (DSM IV defined) on the IDD. Overall, there was moderate agreement between treating doctors' diagnosis of depression and patients' self-report of depressive symptoms. A multivariate model indicated that being in a current relationship was associated with lowered odds of depression (OR = 0.43; CI = 0.23-0.81). The factors strongly associated with increased odds of depression were a past history of illicit drug use (OR = 2.98; CI = 1.60-5.54) and a diagnosis of 'stress' by treating doctors (OR = 5.65; CI = 2.50-12.77). HIV-related medical variables such as immune function, use of antiretro-viral medication and duration of HIV infection were not associated with depression. CONCLUSIONS: There was a high rate of self-reported depression in this group of PLWHA which was also recognized by treating clinicians. Being in a relationship appeared to afford protection against depression while having a history of illicit drug use and current 'stress' were highly associated with depression. Interestingly, HIV-related medical variables including laboratory markers of HIV disease, duration of illness and antiretroviral medication regimen were not related to depression. PMID- 12534661 TI - The association between depression and isotretinoin use in acne. AB - OBJECTIVE: The association between isotretinoin and depression has received little attention in the psychiatric literature despite an increasing number of reports in medical journals. The purpose of this paper is to highlight this association, examine the possible link and review the clinical implications. METHOD: A critical review of the literature pertaining to depression in patients with acne who were treated with isotretinoin was conducted. RESULTS AND CONCLUSIONS: The causal relationship between isotretinoin therapy and depression has not been clearly established and needs further study. Isotretinoin is likely to have a positive psychological impact for the majority of patients who benefit from such a highly efficacious anti-acne treatment. However, it is important to recognize that depression can occur as an idiosyncratic side-effect that requires urgent and appropriate treatment. Therefore, having a low threshold for detection of this uncommon complication and early psychiatric referral to address both the depression and its contributing factors may prevent serious consequences. PMID- 12534662 TI - A survey of the quality of web based information on the treatment of schizophrenia and Attention Deficit Hyperactivity Disorder. AB - OBJECTIVE: To systematically assess the quality, accountability and readability of Internet information on the treatment of schizophrenia and Attention Deficit Hyperactivity Disorder (ADHD), using a standardized pro forma. METHOD: We analysed the 20 most highly ranked pages on the treatment of ADHD and schizophrenia, identified by five common Internet search engines. RESULTS: There was little overlap in the sites identified by different search engines. In the case of schizophrenia, one site was identified three times and another eight sites twice; while for ADHD four sites were identified twice. Accountability (Silberg score), presentation and readability, as assessed by the Flesch-Kincaid Grade Level score, were poor. Mean Silberg, presentation and Flesch-Kincaid Grade Level scores were 3.2 (range 0-9) out of 9, 1.9 (range 0-4) out of 4, and 11.5 (range 6.5-12.25), respectively. There was no statistical difference in scores between the two diagnoses. Depending on the recommendation, agreement with evidence-based practice for schizophrenia ranged from only 2 to 55% (mean = 2.8 (range 0-9) out of 12), while that for ADHD was from 14 to 54% (mean = 1.6 (range 0-6) out of 6). Only 50% of the sites advised readers to clarify information with an appropriate health professional. Interrater reliability in pro forma scores for schizophrenia and ADHD was high (r = 0.96 and 0.95, respectively, p < 0.0001). Sites in the top 10% of scores were significantly more likely to be owned by an organization or have an editorial board than those in the bottom 10%. CONCLUSIONS: The Internet contains misleading information on both schizophrenia and ADHD. The methodology used in this paper could be adapted for other psychiatric conditions. PMID- 12534663 TI - Valid assessment of the clinical features of depression by relatives appears to slip under the RADAR. AB - OBJECTIVE: We report on the development of an observational measure designed for completion by relatives, the Recent Appearance of Depression Assessed by Relatives (RADAR), and consider its validity. METHOD: One hundred and one patients with a current major depressive episode had a relative or close friend complete the RADAR, while psychiatrists and research assistants collected extensive data. RESULTS: Correlated against both patient self-report and psychiatrist-rated depression severity measures, RADAR scores evidenced poor validity overall. Recent Appearance of Depression Assessed by Relatives scores also failed to differentiate depressive subtypes, in that scores for patients with melancholic depression were not significantly higher than for those with non melancholic depression. CONCLUSIONS: Results are consistent with previous findings of poor agreement between clinicians and corroborative witnesses in assessing clinical depressive features, and argue against reliance on corroborative witness reports. PMID- 12534665 TI - Clinical and personality correlates of a new measure of depression: a general practice study. AB - OBJECTIVE: There is a need for a brief, efficient depression screening measure for general practice settings, particularly for identifying those who are at high risk of depression. We therefore test the usefulness of a measure developed in a sample of medically ill hospitalized patients. METHOD: More than 600 patients attending six Sydney general practices completed the Depression in the Medically Ill (DMI-10) measure, in conjunction with sociodemographic, depression history and personality profile measures. The impacts of sociodemographic, personality and lifetime depression variables on DMI-10 scores (and identified 'cases') were examined as a measure of its usefulness. RESULTS: Gender did not influence depression scores, while there were slight associations between DMI-10 scores and age, marital and occupational status. Higher scores were returned by those with more severe, lengthier and perceived stressful medical illnesses. Using a predetermined cut-off score, 36% rated as putative 'cases', a prevalence almost identical to our general hospital study. 'Cases' were distinctly more likely to have had previous depressive episodes, to have sought help for such episodes and to have received antidepressant medication. They also scored higher on measures of anxious (anxious worrying and irritability) and self-critical ('depressive personality') personality styles. CONCLUSIONS: The DMI-10 appears useful as a brief and acceptable screen for depression in a general practice setting, both identifying those who are likely to be currently depressed and those with a background of previous depression. PMID- 12534664 TI - Establishment of a disaster-related psychological screening test. AB - OBJECTIVE: To create a short screening scale for the detection of posttraumatic stress disorder (PTSD) and major depressive disorder (MDE) in earthquake survivors in Taiwan. METHOD: Trained research assistants used the Disaster Related Psychological Screening Test (DRPST) to assess 461 residents of a village that had experienced a major earthquake. The participants were also evaluated by psychiatrists using the Mini-International Neuro-psychiatric Interview (MINI). Best subset regression analysis and the receiver operating characteristics curve were used to select a subset of items and cut-off points from the DRPST. RESULTS: A seven-symptom scale and a three-symptom analogue were selected for PTSD and MDE screening, respectively. Scores of three or more on the PTSD scale and two or more on the MDE scale were used to define a group of positive cases that provide useful information for the patient cohort and will be valuable in long-term follow-up studies of the prevalence of psychiatric diseases following a natural disaster. However, higher scores could also be used to define positive cases under limited psychiatric care resources. CONCLUSION: The DRPST, which was administered for phase 1 of this two-phase study, may be used for effective and rapid screening for PTSD and MDE after an earthquake, despite the usual limitations on resources following a disaster. PMID- 12534666 TI - Mass media and suicide. PMID- 12534667 TI - Psychiatric intensive care units. PMID- 12534668 TI - The clinical profile of psychotic depression. PMID- 12534670 TI - Razing Ockham. PMID- 12534669 TI - The use of intramuscular midazolam in an acute psychiatric unit. PMID- 12534671 TI - Reply to Dr Mahendran. We have not found reported cases of [obsessive-compulsive] OC arising from de novo with olanzapine treatment. PMID- 12534672 TI - Memoriam to Dr George Engel. PMID- 12534705 TI - Third Japan-Australia cardiovascular bioengineering symposium. PMID- 12534706 TI - Recent progress in artificial organ research at Tohoku University. AB - Tohoku University has developed various artificial organs over the last 30 years. Pneumatic driven ventricular assist devices with a silicone ball valve have been designed by the flow visualization method, and clinical trials have been performed in Tohoku University Hospital. On the basis of these developments, a pneumatic driven total artificial heart has been developed and an animal experimental evaluation was conducted. The development of artificial organs in Tohoku University has now progressed to the totally implantable type using the transcutaneous energy transmission system with amorphous fibers for magnetic shielding. Examples of implantable systems include a vibrating flow pump for ventricular assist device, an artificial myocardium by the use of shape memory alloy with Peltier elements, and an artificial sphincter for patients with a stoma. An automatic control system for artificial organs had been developed for the ventricular assist devices including a rotary blood pump to avoid suction and to maintain left and right heart balance. Based upon the technology of automatic control algorithm, a new diagnostic tool for evaluating autonomic nerve function has been developed as a branch of artificial organ research and this new machine has been tested in Tohoku University Hospital. Tohoku University is following a variety of approaches aimed at innovation in artificial organs and medical engineering fields. PMID- 12534707 TI - Korean artificial heart (AnyHeart): an experimental study and the first human application. AB - A Korean artificial heart (AnyHeart) has been implanted in 29 various animals (52 470 kg) to evaluate hemodynamic performance and electromechanical stability. Most were implantable biventricular assist devices in use. A right thoracotomy approach has been a standard technique of implantation. A preclinical fitting test was also performed to observe anatomical feasibility and to compare surgical techniques in 10 human cadavers. The first case of human application was made as a lifesaving procedure on June 12, 2001. PMID- 12534708 TI - Total artificial heart: the adaptation and pathophysiological deviations in the recipient. AB - In the short historical review the principal technical and pathophysiological approaches of the Czech total artificial heart (TAH) research are emphasized. Survival was endangered by arterial and venous hypertension, especially the central venous pressure (CVP) increase, mineralization of driving diaphragms, thromboembolization, and infection. By the appropriate combination of antihypertensive, antimineralization, and anticoagulation treatment and of measures against infection, the survival of the experimental calves was increased (the longest one was 314 days of pumping). Technical and constructional improvement of the blood pumps was another cause of successful experiments. All details are issued in the references. PMID- 12534709 TI - Initial in vivo experience of the VentrAssist implantable rotary blood pump in sheep. AB - VentrAssist (VentrAssist Division, Ventracor Ltd., Chatswood, NSW, Australia) has developed an implantable centrifugal blood pump with an integrated rotor and impeller that is hydrodynamically suspended. Bench testing has been used to assess the performance of the pump under a broad range of operating conditions. This study examined the performance of the pump in vivo up to 90 days implantation. Pumps were implanted via a left lateral thoracotomy. The inflow cannula was inserted at the apex of the left ventricle. The outflow cannula was anastomosed to the descending thoracic aorta. Eighteen implants were performed. Poor recovery from surgery was the main cause of early study termination. These studies demonstrate the suitability of the animal model for evaluation of the VentrAssist rotary blood pump. Further in vivo studies prior to preclinical trials are in progress. PMID- 12534710 TI - Progress in the control system of the undulation pump total artificial heart. AB - The undulation pump total artificial heart (UPTAH) is a small implantable total artificial heart. As the UPTAH generates outflow and inflow at the same time, control of the UPTAH is very difficult. Therefore suitable control methods specifically for the UPTAH should be established. Various motor control, left right flow balance control, and physiological control methods were examined and tried for the UPTAH control in this study. The control system is divided into seven categories. It has a hierarchical structure and all control modes work at the same time. The UPTAH with the newly developed control method has been implanted into the chest cavities of 48 goats. Until now, six goats survived for more than one month, including 63 days in the longest case. The good condition of the UPTAH implanted animal could be maintained with the newly developed control scheme, consisting of the 1/R control and several other additional controls. PMID- 12534711 TI - Computational fluid dynamics analysis of an intra-cardiac axial flow pump. AB - A low rate of hemolysis is an important factor for the development of a rotary blood pump. It is, however, difficult to identify the areas where hemolysis occurs. Computational fluid dynamics (CFD) analysis enables the engineer to predict hemolysis on a computer. In this study, fluid dynamics throughout intracardiac axial flow pumps with different designs were analyzed three dimensionally using CFD software. The computed pressure-flow characteristics of the pump were in good agreement with the measurements. The Reynolds shear stress was computed along particle trace lines. Hemolysis was estimated on the basis of shear stress (tau) and its exposure time (Deltat): dHb/Hb = 3.62 x 10( 7)(tau)(i)(2.416) x Delta(t)(i)(0.785). Particle damage increased with time along the particle trace lines. Hemolysis of each of the pumps was measured in vitro. The computed hemolysis values were in good agreement with the experimental results. CFD is a useful tool for developing a rotary blood pump. PMID- 12534712 TI - Computational flow visualization in vibrating flow pump type artificial heart by unstructured grid. AB - Computational flow visualization in the casing of vibrating flow pump (VFP) was made for various conditions based on the novel techniques of fluid dynamics. VFP type artificial heart can generate the oscillated flow and can be applied to the left ventricular assist device. Flow pattern of blood in an artificial heart is closely connected to mechanical performance and serious biomechanical problems such as hemolysis and blood coagulation. To effectively design the VFP for a left ventricular assist device, the numerical codes for solving Navier-Stokes equations were developed for three-dimensional blood flow based on the finite volume method. Furthermore, the simulation techniques based on the artificial compressibility method and the unstructured grid were also developed here. The numerical calculations were based on the precise configurations and the flow conditions of the prototype device. From the viewpoint of computational fluid dynamics (CFD), the detailed discussion of flow patterns in the casing of VFP, which were closely connected with hemolysis and blood coagulation, was made and the computational results were visualized by the use of the recent technique of computational graphics. Some useful design data of VFP were presented. PMID- 12534713 TI - Numerical analysis of the three-dimensional blood flow in the korean artificial heart. AB - Flow in the blood sac of the Korean artificial heart is numerically simulated by finite element method. Fluid-structure interaction algorithm is employed to compute the three-dimensional blood flow interacting with the sac material. For verification of the numerical method of fluid-structure interaction, two dimensional flow in a collapsible channel with initial tension is simulated and the results are compared with numerical solutions from the literature. Incompressible viscous flow and linear elastic solid are assumed for the blood and the sac material in the device, respectively. The motion of the actuator is simplified by a time-varying pressure boundary condition imposed on the outer surface of the sac. Numerical solutions on the unsteady three-dimensional blood flow in the sac are provided for the cactus-type model in this study. During systole, the inlet is closed and the blood sac is squeezed by the action of the prescribed pressure on the surface. During diastole, the sac is filled with the blood coming from the inlet while the outlet is closed. A strong flow to the outlet and a stagnated flow near the inlet are observed during systole. Shear stress distribution is also delineated to assess the possibility of thrombus formation. We also simulate numerically the hemodynamics of "the reversed model" where the inlet and outlet are reversed for surgical convenience. It is observed that a recirculating flow was generated near the inner corner of the sac in the reversed model. To assess the material strength of the sac, the shear stress distribution in the solid material is also presented. PMID- 12534714 TI - Blood compatible design of a pulsatile blood pump using computational fluid dynamics and computer-aided design and manufacturing technology. AB - Thrombus formation is a critical issue when designing a long-term implantable left ventricular assist system (LVAS). Fluid dynamic characteristics of blood flow are one of the main factors that cause thrombus formation. In this study, we optimized the fluid dynamics of a sac blood pump in our LVAS to ensure minimization of shear-related blood damage that could lead to thrombus formation. A pump housing and a sac chamber were designed with computer-aided design (CAD) software, and fluid dynamics were estimated by computational fluid dynamic (CFD) analysis. We adopted distribution of CFD results for qualitative evaluation, and we also tried to estimate normalized index of hemolysis (NIH) from the results of CFD analysis as a quantitative index of optimization for geometry of the blood pump chamber. A prototype model of the optimized blood pump was made using a three-axis computer machine tool by whittling pieces of nonfoamed polyurethane. Shear stress and theoretical NIH in the redesigned model were lower than those in the first model. Area of flow stagnation that was observed in the first model was not seen in the redesigned model. The results demonstrate that application of CAD/CAM technology to design an artificial heart contributes to optimizing a blood pump chamber for the purpose of reducing thrombus formation. PMID- 12534715 TI - A study on an energy supply method for a transcutaneous energy transmission system. AB - This study proposes a new type of a transcutaneous energy transmission system (TETS) that can supply electrical power for an implanted device without an external battery. In this system, the power is supplied from the floor to the shoes of the patients through coils that are set beneath the floor and the bottom of the shoes. If the patients wear the special shoes, they will be able to move freely on the specially designed floor without an external battery. Direct current (DC)-DC power efficiency was measured in the experiments, and the results showed that it varies with relative positions between the shoe and the floor coils. The results suggested that three-layered floor coils would enable the system to meet the demand for providing the required power anywhere on the floor without intermission. DC-DC power efficiency could be kept over 60% under the practical condition. It can then be concluded that the proposed system has a potential to provide better quality of life for the patients using a TETS. PMID- 12534716 TI - Miniature vibrating flow blood pump using a cross-slider mechanism for external shunt catheter. AB - The prototype of the miniature vibrating flow pump (VFP) is developed for the external shunt catheter. The cross-slider mechanism is applied to vibrate the tube, which causes the pumping effect. This mechanism results in successful development of the miniature and lightweight VFP. By the use of the prototype VFP, the experiment of the basic pump performance is made in detail based on the authorized procedure in the research field of fluids engineering. The typical H-Q curve of VFP, which is the relationship between the pump head and the flow rate, can be obtained. This result suggests that the miniature VFP developed here can be expected to be used as the booster pump for the external shunt catheter in clinical applications. PMID- 12534717 TI - Development of a polymer bileaflet valve to realize a low-cost pulsatile blood pump. AB - The final goal of this study is to realize a low-cost pulsatile blood pump especially for patients with acute heart failure or postoperative low cardiac output syndrome. In support of the pump, two types of polymer bileaflet valves with different configuration of the valve seats were developed. Influence of the leaflet thickness on the hydrodynamics of the prototype was preliminarily investigated among 70 microm, 100 microm, and 150 microm. As to the valves with the thinner leaflets, buckling of the leaflets was observed, which induced a large amount of regurgitation at valve closure. However, by thickening the leaflet to 150 microm, the mean flow of the prototype and the second model could be successfully comparable to the Medtronic-Hall valve. Moreover, accelerated fatigue tests showed that reinforcement of the valve seat with the additional spokes in the second model extended the durability by four times as compared with the prototype, equivalent to an in vivo duration of over one month. PMID- 12534718 TI - Investigation on the mechanical properties of contracted collagen gels as a scaffold for tissue engineering. AB - In this article the mechanical properties of contracted collagen gels were investigated thoroughly by means of uniaxial tensile test. Large type I collagen Dulbecco's Modified Eagle Medium (DMEM) gels (each was 26 ml in volume, 1.67 mg/ml collagen concentration), each populated with about 2.5 x 106 human fibroblasts, were made in 100 mm diameter plastic dishes precoated with albumin for floating the gels in DMEM. Such identically treated gels were divided into three groups for the mechanical measurements at different culture periods (2, 4, and 10 weeks). Rapid contraction occurred within the first 3 days and then the contraction went slowly in the rest period until it reached about 13% of its original size. The stress-strain curve of the contracted collagen gels demonstrated an exponential behavior at low stress region, followed by linear region, a point of yielding, and finally an ultimate stress point at which the maximum stress was reached. The mechanical strength increased in the first few weeks and then decreased as the culture went on. It is obvious that the collagen fibrils formed and were forced to orientate to the tensile direction after the test. The stress relaxation and cyclic creep phenomena were observed. Based on the morphological analysis of transmission electron microscopy (TEM) of the gels, a nonlinear visco-elastic-plastic constitutive formula was proposed, which was able to reproduce the rheological phenomena of the gels. This experiment shows that the human fibroblasts significantly contracted collagen gels so as to achieve certain mechanical strength, which makes it possible to be a scaffold for tissue engineering. However, a further method to reinforce the mechanical strength by several folds must be considered. Meanwhile, the rheological phenomena should be taken into account in the fabrication and application of the structure. PMID- 12534719 TI - Outflow control for avoiding atrial suction in a continuous flow total artificial heart. AB - Continuous flow blood pumps, such as axial flow and centrifugal pumps, have been gaining interest as circulatory devices for total artificial hearts (TAHs) because of their smaller size and simpler structure compared to pulsatile pumps. However, continuous flow pumps are more prone to atrial wall suction than pulsatile pumps are. Sudden increases in flow rate to meet changes in physiological demand, especially in the left pump, often cause atrial wall suction. In this study, a control algorithm to prevent atrial wall suction from occurring in the left atrium by controlling the rotational speed of the right pump, instead of reducing the cardiac output of the left pump, was developed and investigated. The method was tested in a mock circulatory system and in acute animal experiments with adult goats. Two centrifugal pumps were used to totally replace the circulatory function of the natural heart. The cardiac output of each pump was determined independently by a control algorithm running on a computer connected through a serial interface to the pump driving units. Results showed that left atrial wall suction could be prevented using this method, and that the method could be performed simultaneously with physiological control of the artificial heart. PMID- 12534720 TI - In vivo test of pressure head and flow rate estimation in a continuous-flow artificial heart. AB - To avoid using sensors with low biocompatibility and low durability in implantable total artificial heart (TAH) systems, the authors previously proposed a new method for estimating instantaneous values of flow rate and pressure head on the basis of voltage, current, and rotational speed in a motor driven centrifugal pump. The previous in vitro experiments showed that the proposed estimator could automatically compensate for the effect of the change in blood viscosity on the estimation accuracy by employing two kinds of autoregressive exogenous models. In this study, validity and reliability of this estimation method were ascertained in an acute animal experiment. In the experiment, two centrifugal blood pumps were implanted into an adult goat as a total artificial heart. Results of estimation were compared with true values when blood viscosity was changed by injecting physiological saline. The results indicated that the system could successfully estimate pressure head by compensating the change of viscosity, although the estimation accuracy of the in vivo estimation was not so high as that of the previous in vitro tests. PMID- 12534721 TI - Chaos attractors of ventricular elastance to evaluate cardiac performance. AB - Quantitative evaluation of cardiac function is very important in the clinical application of a ventricular assist device. This article reports a new evaluating method of E max, which is the most reliable parameter to evaluate cardiac function. Fluctuation in the E max time series data was evaluated by the nonlinear mathematical analyzing method including chaos and fractal theory. Experimental goats were anesthetized with halothane inhalation, and left ventricular volume and pressure were measured with other hemodynamic parameters to evaluate E max during various drug administrations. E max was evaluated by two methods. One was the conventional pressure volume loop evaluation and the other was the parameter optimization method without left ventricular volume data. As a result, E max evaluated by the parameter optimization method correlated well with the E max with conventional PV curve. Furthermore, interesting results were obtained. There were rhythmical fluctuations in the E max time series data. By the methodology of Takens, E max time series data was embedded into the phase space and a strange attractor was observed. These results may be important when considering E max evaluation during left ventricular assistance. PMID- 12534722 TI - Evaluation of hemolysis in the VentrAssist implantable rotary blood pump. AB - The VentrAssist implantable rotary blood pump (IRBP) is an implantable centrifugal blood pump with a hydrodynamically suspended impeller; optimal efficiency requires small running clearances (70-300 microm). The effect of running clearance and polish on hemolysis was evaluated in vitro. Three different human blood suspensions were compared: phosphate buffered saline (PBS), plasma volume expander (Hemaccel), and whole blood. The test conditions were: blood hematocrit 30%, flow rate 5 L/min, pressure across pump 100 mm Hg, 6 h flow period, and 37 degrees C. Normalized Index of Hemolysis (NIH) for the Biomedicus BP-80, used as a control, was: 0.0040 +/- 0.0023 (n = 9; x +/- SD) and 0.00014 +/ 0.00009 (n = 5) for pooled blood suspensions in PBS and Hemaccel respectively, and 0.00053 +/- 0.0002 (n = 3) in whole blood. Hemolysis was reduced by improved surface finish and unaffected by running clearance. NIH for the VentrAssist IRBP with 0.2 microm Ra surface finish was 0.000167 +/- 0.00007 (n = 4) g/100 L in whole human blood, demonstrating minimal hemolysis. PMID- 12534723 TI - Microvessels of bulbar conjunctiva in UPTAH goats. AB - Aiming to observe directly the microcirculation after total artificial heart (TAH) implantation, we performed a long-term follow-up in 2 goats using conjunctival angioscopy. A short segment of parallel arteriole and venule was photographed and analyzed on computer picture program (magnification 40x). Three main parameters were measured: arteriole diameter, venule diameter, and arteriovenous ratio (A/V ratio). The intrathoracically implanted TAH was the undulation pump total artificial heart (UPTAH) with cardiac output of 100 ml/kg/min. To stabilize the peripheral hemodynamics a 1/R biofeedback control system was used. Our results provided only elementary data about morphology of bulbar microvessels. The main finding was the tendency to general vasoconstriction, more intensive on the venous side (*P < 0.05 in one goat). We did not observe any pathological shapes (e.g., tortuosities, varicosities, or sludge); this result could be attributed to the high effectivity of 1/R control method. These preliminary results should be considered only as an attempt to apply the widely used clinical method of conjunctival angioscopy to the conditions of TAH. PMID- 12534724 TI - Development of totally implantable pulsatile biventricular assist device. AB - Approximately 10% to 15% of all patients implanted with left ventricular assist devices (LVADs) have required right heart support with another device. The necessity of aggressive biventricular support has already been proposed. Therefore, the totally implantable biventricular assist device (BVAD) was developed. The width of the BVAD main body was 87 mm, the thickness 67 mm, and the height 106 mm, while the weight was 785 g. The automatic control algorithm was developed to prevent lung edema and atrial rupture. PMID- 12534727 TI - Fractures of the zygoma. PMID- 12534726 TI - Colonoscopic surveillance after surgery for colorectal cancer: how soon? How often? And how to get the message through? PMID- 12534728 TI - Colonoscopic surveillance after surgery for colorectal cancer. AB - BACKGROUND: Intensive colonoscopic surveillance after resection of colorectal cancer (CRC) has been shown not to improve outcome. The National Health and Medical Research Council of Australia (NHMRC) has recently published guidelines recommending appropriate surveillance intervals after CRC resection. The aims of the present study were to assess current and past patterns of postoperative CRC surveillance and to determine the yield of neoplasia from such surveillance. METHODS: An audit was performed of all patients who underwent colonoscopy following surgical resection of CRC from 1989 to 2001. Two groups were assessed: (i) all patients undergoing surveillance colonoscopies; and (ii) all patients diagnosed with CRC at Sir Charles Gairdner Hospital (SCGH) who subsequently had postoperative colonoscopies. Patients who had their index colonoscopy at the study centre and who subsequently underwent surveillance colonoscopies were studied in detail. Yield for neoplasia, patterns of surveillance and concordance with NHMRC recommendations were determined. RESULTS: There were 990 surveillance examinations performed and colorectal adenomas were identified in 184. However, only one case of recurrent cancer was detected. There were a total of 161 patients who had CRC diagnosed and underwent surveillance at SCGH. Of these patients, 75% underwent colonoscopy at 12 months after resection and 48% of these cases underwent a further examination within 12 months. Only 23% of examinations concurred with NHMRC recommendations and practice has not changed with release of these guidelines. CONCLUSIONS: Resectable CRC recurrences are rarely detected at colonoscopic surveillance. Surveillance colonoscopies are -performed too frequently and release of NHMRC guidelines has failed to change practice. PMID- 12534729 TI - Results of resections for hepatocellular carcinoma in a new hepatobiliary unit. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is the fourth commonest cancer in Singapore. Surgical resection offers the only chance of "cure". Resection tends to be difficult in HCC because of late presentation and concomitant liver cirrhosis. Specialized units with higher volume of operations are known to produce better results. The present review aims to evaluate early results with HCC resection and discuss the correlation between various tumour prognostic factors and the outcome. METHODS: The records of 81 consecutive hepatic resections for HCC in the hepatobiliary unit of the Department of General Surgery at the Singapore General Hospital from 1 January 1996 to 31 December 1998 were retrospectively reviewed. RESULTS: The mean age of the patients was 56.0 +/- 15.4 years. There were more men (M:F: 72:9) and Chinese patients (75 Chinese patients (92.6%); four Malay patients (4.9%); two Indian patients (2.5%)) affected by the disease. Hepatitis B and C carrier status were present in 67.1% (n = 51) and 3.9% (n = 1) of the patients, respectively. Forty-two patients (53.2%) had underlying liver cirrhosis. Twenty-eight patients (34.6%) underwent major hepatectomy and 53 (65.4%) underwent minor hepatectomies. Perioperative mortality was 4.9% (n = 4). The morbidity rate following hepatic resections was 28.4% (n = 23). The median follow up was 21 months (range: 1-52 months). The median survival was 43 months (95% confidence interval (CI): 34.6-51.4 months) after surgery and median time to recurrence was 9.6 months (range: 2-32 months). Overall survival was 79% and 59% at 1 and 3 years, respectively. Disease-free survival was 59% and 30% at 1 and 3 years, respectively. Advance pathological tumour, nodes, metastases (TNM) stage (III and IV), and presence of adjacent organ involvement were risk factors for early recurrence. Advance pathological TNM stage (III and IV) and blood loss of more than 2 L were poor prognostic factors for overall survival. CONCLUSION: The results of hepatectomies for HCC in the newly established unit at Singapore General Hospital have been shown to be comparable to other established specialized hepatobiliary units with similar perioperative mortality and morbidity rates. PMID- 12534731 TI - Cutaneous innervation in man before and after lumbar sympathectomy: evidence for interruption of both sensory and vasomotor nerve fibres. AB - BACKGROUND: Rest pain and severe ischaemia in patients who are unable to be offered (further) surgery to revascularize the lower limb is still problematic. Lumbar sympathectomy has been used for many years but the mechanisms by which this works are not absolutely clear. Both sensory and vasomotor fibres travel in the lumbar sympathetic chain and the effects of lumbar sympathectomy on these nerve types have been investigated in the present paper. METHODS: Immunohistochemical methods were used to detect neuropeptides contained in sensory and vasomotor nerves in the lower limb skin of (i) patients having amputations for peripheral vascular disease (PVD) after previous (chemical or surgical) sympathectomy; (ii) patients having amputations for PVD without previous (chemical or surgical) sympathectomy; and in control normal skin. The three groups are compared and the results are discussed. RESULTS: Normal and PVD controls had intact sensory and vasomotor nerves around dermal cutaneous blood vessels, but these were completely or virtually completely lost after lumbar sympathectomy, by either chemical or surgical means. CONCLUSIONS: Lumbar sympathectomy severs both vasomotor and sensory fibres, suggesting that relief of rest pain may be explained not only by increased cutaneous and muscle blood flow, but also by nociceptive sensory denervation. PMID- 12534732 TI - Thoracoscopic talc pleurodesis for malignant pleural effusion. AB - BACKGROUND: Malignant pleural effusion (MPE) is a common and distressing condition at the end of life for many patients with disseminated cancer. The challenge for the surgeon lies in managing this problem in order to deliver the most effective palliation with the least impact on the limited time available to these patients. METHODS: Herein is reported a retrospective review of outcomes for a consecutive series of 66 MPE (61 patients) treated over a 5-year period from 1995 to 2000. A standard operative technique involving a single-lung anaesthetic and two-port thoracoscopy was employed. Outcomes were determined by contacting the referring practitioner or the patients themselves. Principal outcome measures included time to recurrence of the effusion and survival. RESULTS: Complete follow up was achieved for 60 MPE (55 patients; five of whom were treated for metachronous, bilateral disease). The three most common primary sites were breast, lung and mesothelial tissue. The planned procedure was not completed in two cases due to encasement of the underlying lung by tumour. Primary failure (immediate recurrence of the effusion) occurred in six cases. Delayed recurrence of the effusion occurred in a further 23 MPE resulting in complete control in 31 cases (52%) until death. Overall median survival was 220 days and the 30-day mortality was 0. CONCLUSIONS: Complete and permanent control of a malignant effusion is difficult to achieve. Management based on thoracoscopy and talc insufflation produces satisfactory results with an acceptable morbidity and no early mortality. The ability to inspect the pleural space, break down adhesions and completely drain pockets of fluid to achieve complete lung expansion probably contributes to this. PMID- 12534733 TI - Application of image-guided biopsy for impalpable breast lesions in Chinese women. AB - BACKGROUND: Screening for breast cancer has resulted in an increasing number of mammographically detected lesions that require further management. The Advanced Breast Biopsy Instrumentation system is a recently added biopsy technique for the management of such lesions. The present paper will review the authors' experience in the use of this procedure in Chinese patients whose breast volume was smaller than that of Caucasians. METHODS: Ninety-three patients were listed for the procedure and 78 (84%) underwent the procedure successfully. Ninety-two lesions were biopsied. Advanced Breast Biopsy Instrumentation (ABBI) was performed for clustered microcalcifications or abnormal mass/density. Minimally Invasive Breast Biopsy (MIBB), a suction-assisted core biopsy device, was employed for more scattered lesions. For small volume breasts, it may be required to bring the hand through the aperture to get the targeted lesions onto the digital image or, in the case of ABBI, to excise just beyond the deep margin of the lesion rather than the recommended depth. RESULTS: The ABBI was performed for 43 (46.7%) lesions and MIBB for 49 (53.3%) lesions. Nine (9.8%) were diagnosed to have ductal carcinoma in situ, two (2.2%) had ductal carcinoma in situ with microinvasion and eight (8.7%) had invasive ductal carcinoma. All the malignant lesions required further management. In addition, 19 (20.7%) were found to have atypical hyperplasia. Patients' satisfaction and cosmetic outcome are good. CONCLUSION: The ABBI and MIBB procedures can be applied satisfactorily for biopsy of mammographic lesions with good -cosmetic outcome in Chinese patients. PMID- 12534734 TI - Extra-nodal lymphoma presenting as a mimic of soft-tissue sarcoma. AB - BACKGROUND: A variety of soft-tissue tumours result in soft-tissue masses, which are all differential diagnoses of soft-tissue sarcomas. A rare cause is extra nodal lymphoma which, unlike a soft-tissue sarcoma which it mimics, should not be excised, but rather treated by chemotherapy and/or radiotherapy. The aim of the present study was to identify clinical, radiological, biopsy and management features of lymphoma when presenting as a soft-tissue mass. METHODS: A review of 17 cases of soft-tissue lymphoma presenting as a soft-tissue mass from a total database of 295 soft-tissue tumours was undertaken. Lymphomas arising in cervical, axillary and inguinal nodes were excluded. RESULTS: All patients presented with a soft-tissue mass but none had the symptoms of lymphoma. Computed tomography scanning identified seven (54%) of 13 with regional lymphadenopathy, six (46%) of 13 with encasement of major vascular structures and three with invasion across major soft-tissue boundaries. Core biopsy established a diagnosis in 13 patients (sensitivity 93%). All patients were treated with either chemotherapy and/or radiotherapy and all are alive with a mean follow up of 19 months. None underwent major resectional surgery. CONCLUSION: The present series highlights a rare subgroup of patients with extra-nodal soft-tissue lymphoma mimicking soft-tissue sarcoma. Core biopsy is accurate in their diagnosis. Computed tomography scanning may show features suggesting the diagnosis. PMID- 12534735 TI - Temporal trends and clinical correlates for the ret/PTC1 mutation in papillary thyroid carcinoma. AB - BACKGROUND: The incidence of papillary thyroid carcinoma (PTC) has increased in Australia at a rate exceeding 10% per annum over the past two decades. In Tasmania the increase has averaged 24% per year between 1982 and 1997. Exposure to ionizing radiation is the best characterized risk factor for PTC. Oncogenic mutations of the RET proto-oncogene (ret/PTC rearrangements) have been associated with PTC arising following radiation exposure. In the present study it was sought to determine if PTC incidence trends were associated with an increased occurrence of the ret/PTC1 rearrangement. METHODS: All cases of PTC diagnosed in Tasmania during the even numbered years 1978-1998 inclusive were sought for study (n = 98). Archival histopathology blocks for 62 cases were located. The RNA was successfully extracted from 41 tumours and ret/PTC1 status assessed by reverse transcription-polymerase chain reaction. RESULTS: The ret/PTC1 mutation was found in 26 (63%) of PTC. The mean age at diagnosis for ret/PTC1-positive and ret/PTC1 negative tumours was 46.5 +/- 15.46 and 41.9 +/- 13.45 years, respectively. The ret/PTC1 positivity was significantly associated with larger tumour size. However, ret/PTC1 was not associated with an adverse prognosis. The prevalence of tumours positive for ret/PTC1 remained stable over the study period (1978-1998) and did not exhibit birth year or diagnosis year clustering. CONCLUSION: This is the first study to map temporal trends for the prevalence of ret/PTC1 relative to incidence trends for PTC. Although the ret/PTC1 mutation was frequently identified in Tasmanian PTC, there was no clear relationship between ret/PTC1 and recent PTC incidence trends. PMID- 12534737 TI - Palpable parathyroid adenomas presenting as clinical solitary thyroid nodules and cytologically as follicular thyroid neoplasms. AB - BACKGROUND: A palpable parathyroid mass, in a patient with primary hyperparathyroidism, is presumed to be parathyroid carcinoma until proven otherwise, with other less common causes including parathyroid cysts and adenomas. These parathyroid pathologies can be more difficult to interpret with concomitant thyroid disease. METHODS: A retrospective review was undertaken of a series of three patients with palpable parathyroid adenomas mimicking thyroid nodules. RESULTS: Two of three patients had preoperative biochemical evidence of primary hyperparathyroidism. Sestamibi scanning confirmed the presence of parathyroid pathology in one case. Fine-needle aspiration cytology revealed probable follicular neoplasms of the thyroid in all three cases. Only one of three parathyroid adenomas was diagnosed at neck exploration; the others were diagnosed at subsequent histopathology. CONCLUSION: Parathyroid and thyroid disease often occur simultaneously. Concomitant parathyroid pathology should be considered, even in the absence of biochemical and radiological evidence, at neck exploration for thyroid disease. Macroscopic identification of parathyroid disease at neck exploration can be difficult when within the thyroid gland capsule. Cytology and imaging of parathyroid adenomas may, on occasion, mimic follicular thyroid neoplasms. PMID- 12534739 TI - Relationship between elevated preoperative troponin T and adverse outcomes following cardiac surgery. AB - BACKGROUND: The prognostic value of troponin T (TnT) has been demonstrated in patients following a myocardial infarction. There are limited data regarding the prognostic utility of preoperative TnT in patients undergoing cardiac surgery. The aim of the present study was to determine if elevated preoperative TnT is a predictor of more complex recovery outcomes in the cardiac surgical setting. METHODS: A single preoperative TnT measurement was assessed in 696 patients undergoing isolated coronary artery bypass graft surgery. Elevated preoperative TnT levels were classified as > or =0.2 ng/mL. Preoperative, intraoperative, intensive care and postoperative events were prospectively recorded for all patients, and retrospectively reviewed for the present study. RESULTS: Elevated preoperative TnT levels were detected in 10% (71/696) of patients. Compared to patients with normal TnT levels, elevated preoperative TnT increased the risk of mortality at 30 days (7% vs 1%, P = 0.004, odds ratio (OR) = 6.7) and 2 years (14% vs 3%, P < 0.001, OR = 5.0), and resulted in prolonged intensive care unit (ICU) stays (P < 0.001) and longer postoperative hospitalization (P < 0.001). Elevated preoperative TnT was also associated with an increased need for perioperative and postoperative cardiovascular support, early ischaemic change and postoperative congestive cardiac failure. In multivariate analyses preoperative TnT was a significant independent predictor of 30-day and 2-year mortality, and duration of ICU stay. CONCLUSIONS: Elevated preoperative TnT highlights a subgroup of cardiac surgical patients who are more likely to have a post-operative course with increased morbidity and mortality. PMID- 12534740 TI - Chemical injuries: the Tasmanian Burns Unit experience. AB - BACKGROUND: Chemical burns account for relatively few admissions to a burns unit. These injuries, however, deserve separate consideration because of their ability to cause continuing tissue destruction, their potential to cause systemic toxicity and the value of early treatment with copious lavage. Widespread inexperience in the treatment of chemical burns highlights the potential for greater levels of general awareness and knowledge. METHODS: A review of 31 patients with chemical injuries admitted to the Tasmanian Burns Unit at the Royal Hobart Hospital (RHH) was carried out for the years 1989-1999. RESULTS: The majority of patients were men aged 20-49 years (mean age: 32 years). Fifty-one per cent of injuries occurred in a domestic and 38% in an industrial setting. The more common aetiological agents were cement (25%), sulphuric acid (16%) and hydrofluoric acid (16%). The upper and lower extremities were involved in all but four patients and the mean total body surface area affected was 3.4%. The mean length of hospital stay was 9 days with a range of 1-30 days. Management of injuries consisted of either surgical or conservative treatment. The former included debridement and split-thickness skin grafting or primary closure and the latter of topical treatment with 1% silver sulfadiazine cream and appropriate dressings. CONCLUSION: Widespread inexperience in the treatment of chemical injuries highlights the potential for greater levels of knowledge. This is particularly apparent in the early management of these injuries. PMID- 12534741 TI - Fractured zygomas. AB - BACKGROUND: The purpose of this paper is to provide a review, based on collected data, on the topic of "fractured zygoma". The review is presented under the headings of epidemiology, fracture patterns, treatment modalities and complications. Throughout the paper comparison is made with published data from around the world. METHODS: A 10-year retrospective audit was undertaken of all hospitalized patients, at the Royal Hobart Hospital, Tasmania, who had sustained a fractured zygoma. All Le Fort fractures involving the zygoma were excluded. RESULTS: A total of 263 fractures was sustained, largely due to assault. Alcohol was a significant contributing factor. Tetrapod fractures were the most frequent type of fractures witnessed. Plating was the most frequently employed fixation. Inferior orbital nerve dysfunction and other complications were seen in 24.6% and 20.7% of follow-up cases, respectively. Open reductions were nearly 4 times more likely to be accompanied by complications if inferior orbital nerve dysfunction was excluded. CONCLUSIONS: Both closed and open reductions are good treatment modalities and were used in near equal numbers of patients. There is a higher incidence of postoperative facial deformity in the closed reduction group, but more complications related to the incisions in the open reduction group. Open reduction and internal fixation is advocated for the unstable, markedly displaced or comminuted fractures. Silastic sheeting is a favoured graft for repair of the associated orbital floor defects and is associated with few complications. PMID- 12534742 TI - Retrospective review of paediatric patients with acute scrotum. AB - BACKGROUND: The purpose of the present study was to evaluate the historical features and physical examination findings in boys presenting with acute scrotum and to determine the reliability of these modalities to make the correct clinical diagnosis. METHODS: A retrospective review of all boys presenting with acute scrotal/testicular pathology between January 1994 and December 1998 was performed. RESULTS: Two hundred and four boys were included in the study: 110 with torsion of the testicular appendage (TA), 40 with testicular torsion (TT), 29 with epididymo-orchitis (EO) and 25 with other diagnoses. Seventeen boys were managed conservatively. With the exception of symptom duration, there was no statistically significant difference between boys with TA, TT and EO. Boys with TT sought medical attention significantly earlier (median: 9.5 h) than those with TA (median: 48 h). Boys requiring an orchidectomy (n = 8) presented later than those who did not (median 30 vs 9 h; P < 0.05): seven (88%) were under 7 years of age. The testicular salvage rate was 88% and 36% with a pain duration of <12 h and >12 h, respectively. The preoperative clinical diagnosis was correct in 80% of those boys who underwent surgical exploration. Testicular torsion was misdiagnosed clinically in five boys. Late testicular atrophy was not observed in the present study. CONCLUSIONS: Most boys presenting with acute scrotum have TA. The history and physical examination can reliably identify those boys who can be managed conservatively. Routine surgical exploration is no longer justified in all boys with acute scrotum. PMID- 12534743 TI - Immunological findings in acute and chronic pancreatitis. AB - BACKGROUND: In order to identify the role of mononuclear cells in pancreatitis, the immune picture at the cellular level has been studied. The cytokine production at the individual cell level in response to in vitro stimulation can help in identifying the percentage of specific cell types producing a particular cytokine. METHODS: A total of 30 patients with acute pancreatitis (AP; n = 15) and chronic pancreatitis (CP; n = 15) were entered into the study and peripheral blood samples were drawn at the time of admission. A control group of 12 healthy individuals (age and sex matched) were also included in the study. Mononuclear cells from the peripheral blood from all three groups were separated on Ficoll gradient and labelled for surface antigens (i.e. cluster of differentiation (CD)3, CD4, CD8, CD25, CD14 and human leucocyte antigen D receptor (HLA-DR)) using monoclonal antibodies. The CD3+ and CD14+ cells were also stained for intracellular cytokines by specific monoclonal antibodies (i.e. for interferon (IFN)-gamma, interleukin (IL)-2, IL-4 & IL-10 in CD3+ cells and IL-6 and IL-12 in CD14+ cells). These fluorochrome-labelled cells were analysed on the flow cytometer. RESULTS: The T-cell population is affected in pancreatitis where the CD4+ and CD8+ T-cell numbers are significantly altered. There is a marked reduction in CD4+ T cells in AP and CP. The CD8+ T cells were significantly reduced in AP. The IL-2 receptor is expressed in large numbers in AP and CP patient groups. The T cells from the CP group had a typical IL-2, IL-4 secreting pattern, while AP patients had a high IL-10, IFN-gamma and low IL-2, IL-4 population. The HLA-DR expression on monocytes differed significantly between the two groups, being strikingly reduced in chronic cases and significantly reduced in acute cases. The percentage of IL-6-producing monocytes was significantly higher in patients with AP as well as CP while IL-12 levels were higher in the CP than in the AP group. CONCLUSIONS: T cells in CP are polarized towards the T helper (Th)1 phenotype while the AP patients had a mixed population. This seems to be a part of immune deviation that is associated with development of CP and IL 12 appears to be instrumental in this process. The monocytes synthesize proinflammatory cytokines, which play an important role in the local and systemic consequences of the disease. In the disease clinical assessment such a study can provide useful information about the involvement of the immuno-inflammatory system. PMID- 12534744 TI - Surgical rotations in provincial South Australia: the trainees' perspective. AB - BACKGROUND: Rural general surgery is faced with a shortage of resident surgeons in many parts of Australia. Although it is accepted that an undergraduate rural exposure favourably influences graduates to undertake rural practice, it is not known whether postgraduate terms exert a similar effect. METHOD: Advanced general surgical trainees in 2000 were rotated for 1-month terms to Whyalla, a major provincial centre in South Australia. The trainees were asked to complete a questionnaire before and after the rotation. RESULTS: A total of nine trainees completed a rural term in Whyalla. Eight questionnaires were returned prior to the rotation and seven following the time in Whyalla. The overall experience, and the teaching standards and pathology experienced were rated highly but the term had little effect in changing trainee's attitudes towards eventual practice location. CONCLUSION: Postgraduate surgical terms in South Australia are a relatively new phenomenon compared to other states in Australia. Without a foundation in rural surgery at an undergraduate level, surgical terms for trainees, despite being of high quality, might not be very successful in influencing graduates to practise surgery in rural locations. PMID- 12534745 TI - Melbourne vascular surgical association audit. AB - BACKGROUND: The formation of the Melbourne Vascular Surgical Association has led to the establishment of a vascular surgical audit programme that commenced in January 1999. This has allowed establishment of a benchmark for quality assurance in vascular surgery in Australia. METHODS: A consultative process allowed widespread adoption of the audit across all public hospital vascular units in Melbourne and the two largest regional centres in Victoria. Data were collected at two points during admission: at operation and at discharge. Risk stratification, using logistic regression and risk-adjusted ratios for adverse events was assessed for comparison of outcomes between units for the first 3 years of data collection. There is regular contact with all participants for data feedback and quality control. RESULTS: The standard of vascular surgery across Victoria is consistent, and there has been excellent compliance by all academic vascular units. Private practice data are less complete, and only half of the vascular surgeons have participated. CONCLUSIONS: A statewide audit process is feasible and viable. Coordination by the Melbourne Vascular Surgical Association is crucial for its continued success. PMID- 12534747 TI - The Internet: from basics to telesurgery. PMID- 12534746 TI - Adrenal-sparing surgery for hereditary phaeochromocytoma. PMID- 12534748 TI - Hydatid disease of the liver in pregnancy. PMID- 12534749 TI - Selective use of preoperative endoscopic retrograde cholangiopancreatography in the era of laparoscopic cholecystectomy. PMID- 12534750 TI - Obstructing sigmoid cancer with local invasion in an incarcerated inguinal hernia. PMID- 12534751 TI - Embolization for right hepatic artery pseudoaneurysm following laparoscopic cholecystectomy. PMID- 12534752 TI - External pneumatocoele of the mastoid system. PMID- 12534753 TI - Bilateral brachial plexus injury during laparoscopic sigmoid colectomy. PMID- 12534757 TI - Effect of primary medical care on addiction and medical severity in substance abuse treatment programs. AB - OBJECTIVE: To examine whether the availability of primary medical care on-site at addiction treatment programs or off-site by referral improves patients' addiction severity and medical outcomes, compared to programs that offer no primary care. DESIGN: Secondary analysis of a prospective cohort study of patients admitted to a purposive national sample of substance abuse treatment programs. SETTING: Substance abuse treatment programs in major U.S. metropolitan areas eligible for demonstration grant funding from the federal Substance Abuse and Mental Health Services Administration. RESPONDENTS: Administrators at 52 substance abuse treatment programs, and 2,878 of their patients who completed treatment intake, discharge, and follow-up interviews. MEASUREMENTS: Program administrators reported whether the program had primary medical care available on-site, only off site, or not at all. Patients responded to multiple questions regarding their addiction and medical status in intake and 12-month follow-up interviews. These items were combined into multi-item composite scores of addiction and medical severity. The addiction severity score includes items measuring alcohol and drug use, employment, illegal activities, legal supervision, family and other social support, housing, physical conditions, and psychiatric status. The medical severity score includes measures of perceived health, functional limitations, and comorbid physical conditions. MAIN RESULTS: After controlling for treatment modality, geographic region, and multiple patient-level characteristics, patients who attended programs with on-site primary medical care experienced significantly less addiction severity at 12-month follow-up (regression coefficient, -25.9; 95% confidence interval [95% CI], -43.2 to -8.5), compared with patients who attended programs with no primary medical care. However, on-site care did not significantly influence medical severity at follow-up (coefficient, -0.28; 95% CI, -0.69 to 0.14). Referral to off-site primary care exerted no detectable effects on either addiction severity (coefficient, -9.0; 95% CI, -26.5 to 8.5) or medical severity (coefficient, -0.03; 95% CI, -0.37 to 0.44). CONCLUSIONS: On site primary medical care improves substance abuse treatment patients' addiction related outcomes, but not necessarily their health-related outcomes. Further study is needed to discern the mechanism through which on-site primary care might improve the addiction-related outcomes of substance abuse treatment. PMID- 12534759 TI - Evaluation and management of osteoporosis following hospitalization for low impact fracture. AB - OBJECTIVE: To evaluate the pattern of osteoporosis evaluation and management in postmenopausal women who present with low-impact (minimal trauma) fracture. DESIGN: Retrospective chart review of patients admitted with a fracture in the absence of trauma or bone disease. Telephone follow-up survey was conducted at 12 months after discharge to collect information on physician visits, pharmacological therapies for osteoporosis, functional status, and subsequent fractures. PATIENTS/PARTICIPANTS: Postmenopausal women admitted to a hospital in St. Paul, Minnesota between June 1996 and December 1997 for low-impact fractures were identified. Low-impact fracture was defined as a fracture occurring spontaneously or from a fall no greater than standing height. Retrospective review of 301 patient medical records was conducted to obtain data on pre admission risk factors for osteoporosis and/or fracture, and osteoporosis-related evaluation and management during the course of hospitalization. Follow-up 1 year after the incident fracture was obtained on 227 patients. MEASUREMENTS AND MAIN RESULTS: Two hundred twenty-seven women were included in the study. Osteoporosis was documented in the medical record in 26% (59/227) of the patients at hospital discharge. Within 12 months of hospital discharge, 9.6% (22/227) had a bone mineral density test, and 26.4% (60/227) were prescribed osteoporosis treatment. Of those who were prescribed osteoporosis treatment, 86.6% (52/60) remained on therapy for 1 year. Nineteen women suffered an additional fracture. Compared to women without a prior fracture, women with at least 1 fracture prior to admission were more likely to have osteoporosis diagnosed and to receive osteoporosis related medications. CONCLUSION: Despite guidelines that recommend osteoporosis evaluation in adults experiencing a low-trauma fracture, we report that postmenopausal women hospitalized for low-impact fracture were not sufficiently evaluated or treated for osteoporosis during or after their hospital stay. There are substantial opportunities for improvement of care in this high-risk population to prevent subsequent fractures. PMID- 12534758 TI - Effectiveness of collaborative care depression treatment in Veterans' Affairs primary care. AB - OBJECTIVE: To compare collaborative care for treatment of depression in primary care with consult-liaison (CL) care. In collaborative care, a mental health team provided a treatment plan to the primary care provider, telephoned patients to support adherence to the plan, reviewed treatment results, and suggested modifications to the provider. In CL care, study clinicians informed the primary care provider of the diagnosis and facilitated referrals to psychiatry residents practicing in the primary care clinic. DESIGN: Patients were randomly assigned to treatment model by clinic firm. SETTING: VA primary care clinic. PARTICIPANTS: One hundred sixty-eight collaborative care and 186 CL patients who met criteria for major depression and/or dysthymia. MEASUREMENTS: Hopkins Symptom Checklist (SCL-20), Short Form (SF)-36, Sheehan Disability Scale. MAIN RESULTS: Collaborative care produced greater improvement than CL in depressive symptomatology from baseline to 3 months (SCL-20 change scores), but at 9 months there was no significant difference. The intervention increased the proportion of patients receiving prescriptions and cognitive behavioral therapy. Collaborative care produced significantly greater improvement on the Sheehan at 3 months. A greater proportion of collaborative care patients exhibited an improvement in SF 36 Mental Component Score of 5 points or more from baseline to 9 months. CONCLUSIONS: Collaborative care resulted in more rapid improvement in depression symptomatology, and a more rapid and sustained improvement in mental health status compared to the more standard model. Mounting evidence indicates that collaboration between primary care providers and mental health specialists can improve depression treatment and supports the necessary changes in clinic structure and incentives. PMID- 12534760 TI - Are older patients more satisfied with hospital care than younger patients? AB - OBJECTIVE: Determine relationships between age, self-reported health, and satisfaction in a large cohort of hospitalized patients. DESIGN: Cross-sectional survey. SETTING: Thirty-one hospitals in a large Midwestern metropolitan area. PATIENTS/PARTICIPATION: Randomly selected medical and surgical patients (N = 64,900; mean age, 61 years; 56% female; 84% white) discharged during specific time periods from July 1990 to March 1995 who responded to a mailed survey (overall response rate, 48%). MEASUREMENTS AND MAIN RESULTS: Patients' overall ratings of hospital quality and satisfaction with 5 aspects of care (physician care, nursing care, information provided, discharge instructions, and coordination of care) were measured by a validated survey, which was mailed to patients after discharge. Analyses compared satisfaction in 5 age groups (18 to 35, 36 to 50, 51 to 65, 66 to 80, and > 80 years). Scores for the 5 aspects of care initially increased with age (P <.001) and then declined (P <.001). A similar relationship was found in analyses of the proportion of patients who rated overall quality as "excellent" or "very good." Satisfaction was also higher in patients with better self-reported health (P <.001). In analyses of patients with poor to fair health, satisfaction scores peaked at age 65 before declining. However, for patients with good to excellent health, scores peaked at age 80. Moreover, declines in satisfaction in older patients were lower in patients with better health. These findings were consistent in multivariable analyses adjusting for potential confounders. CONCLUSIONS: Satisfaction exhibits a complex relationship with age, with scores increasing until age 65 to 80 and then declining. This relationship was consistent across individual satisfaction scales, but was modified by health status. The results suggest that age and health status should be taken into account when interpreting patient satisfaction data. PMID- 12534761 TI - Improving physicians' knowledge of the costs of common medications and willingness to consider costs when prescribing. AB - OBJECTIVES: To determine the effectiveness of an educational intervention designed to improve physicians' knowledge of drug costs and foster willingness to consider costs when prescribing. DESIGN: Pre- and post-intervention evaluation, using physicians as their own controls. SETTING: Four teaching hospitals, affiliated with 2 residency programs, in New York City and northern New Jersey. PARTICIPANTS: One hundred forty-six internal medicine house officers and attendings evaluated the intervention (71% response rate). Of these, 109 had also participated in a pre-intervention survey. INTERVENTION: An interactive teaching conference and distribution of a pocket guide, which listed the average wholesale prices of over 100 medications commonly used in primary care MEASUREMENTS AND MAIN RESULTS: We administered a written survey, before and 6 months after the intervention. Changes in attitudes and knowledge were assessed, using physicians as their own controls, with Wilcoxon matched-pairs signed-rank tests. Eighty-five percent of respondents reported receiving the pocket guide and 46% reported attending 1 of the teaching conferences. Of those who received the pocket guide, nearly two thirds (62%) reported using it once a month or more, and more than half (54%) rated it as moderately or very useful. Compared to their baseline responses, physicians after the intervention were more likely to ask patients about their out-of-pocket drug costs (22% before vs 27% after; P <.01) and less likely to feel unaware of drug costs (78% before vs 72% after; P =.02). After the intervention, physicians also reported more concern about the cost of drugs when prescribing for patients with Medicare (58% before vs 72% after; P <.01) or no insurance (90% before vs 98% after; P <.01). Knowledge of the costs of 33 drugs was more accurate after the intervention than before (P <.05). CONCLUSION: Our brief educational intervention led to modest improvements in physicians' knowledge of medication costs and their willingness to consider costs when prescribing. Future research could incorporate more high-intensity strategies, such as outreach visits, and target specific prescribing behaviors. PMID- 12534762 TI - Patient preference for physician discussion and practice of spirituality. AB - OBJECTIVE: To determine patient preferences for addressing religion and spirituality in the medical encounter. DESIGN: Multicenter survey verbally administered by trained research assistants. Survey items included questions on demographics, health status, health care utilization, functional status, spiritual well-being, and patient preference for religious/spiritual involvement in their own medical encounters and in hypothetical medical situations. SETTING: Primary care clinics of 6 academic medical centers in 3 states (NC, Fla, Vt). PATIENTS/PARTICIPANTS: Patients 18 years of age and older who were systematically selected from the waiting rooms of their primary care physicians. MEASUREMENTS AND MAIN RESULTS: Four hundred fifty-six patients participated in the study. One third of patients wanted to be asked about their religious beliefs during a routine office visit. Two thirds felt that physicians should be aware of their religious or spiritual beliefs. Patient agreement with physician spiritual interaction increased strongly with the severity of the illness setting, with 19% patient agreement with physician prayer in a routine office visit, 29% agreement in a hospitalized setting, and 50% agreement in a near-death scenario (P <.001). Patient interest in religious or spiritual interaction decreased when the intensity of the interaction moved from a simple discussion of spiritual issues (33% agree) to physician silent prayer (28% agree) to physician prayer with a patient (19% agree; P <.001). Ten percent of patients were willing to give up time spent on medical issues in an office visit setting to discuss religious/spiritual issues with their physician. After controlling for age, gender, marital status, education, spirituality score, and health care utilization, African-American subjects were more likely to accept this time trade off (odds ratio, 4.9; confidence interval, 2.1 to 11.7). CONCLUSION: Physicians should be aware that a substantial minority of patients desire spiritual interaction in routine office visits. When asked about specific prayer behaviors across a range of clinical scenarios, patient desire for spiritual interaction increased with increasing severity of illness setting and decreased when referring to more-intense spiritual interactions. For most patients, the routine office visit may not be the optimal setting for a physician-patient spiritual dialog. PMID- 12534763 TI - Linguistic and cultural barriers to care. AB - CONTEXT: Primarily because of immigration, Asian Americans are one of the fastest growing and most ethnically diverse minority groups in the United States. However, little is known about their perspectives on health care quality. OBJECTIVE: To examine factors contributing to quality of care from the perspective of Chinese- and Vietnamese-American patients with limited English language skills. DESIGN: Qualitative study using focus groups and content analysis to determine domains of quality of care. SETTING: Four community health centers in Massachusetts. PARTICIPANTS: A total of 122 Chinese- and Vietnamese American patients were interviewed in focus groups by bilingual interviewers using a standardized, translated moderator guide. MAIN OUTCOME MEASURES: Domains of quality of care mentioned by patients in verbatim transcripts. RESULTS: In addition to dimensions of health care quality commonly expressed by Caucasian, English-speaking patients in the United States, Chinese- and Vietnamese-American patients with limited English proficiency wanted to discuss the use of non Western medical practices with their providers, but encountered significant barriers. They viewed providers' knowledge, inquiry, and nonjudgmental acceptance of traditional Asian medical beliefs and practices as part of quality care. Patients also considered the quality of interpreter services to be very important. They preferred using professional interpreters rather than family members, and preferred gender-concordant translators. Furthermore, they expressed the need for help in navigating health care systems and obtaining support services. CONCLUSIONS: Cultural and linguistically appropriate health care services may lead to improved health care quality for Asian-American patients who have limited English language skills. Important aspects of quality include providers' respect for traditional health beliefs and practices, access to professional interpreters, and assistance in obtaining social services. PMID- 12534764 TI - Four simple questions can help screen for eating disorders. AB - Current screening instruments for eating disorders are cumbersome to administer and have not been validated in primary care populations. We compared the performance characteristics of 2 screening tools, the SCOFF clinical prediction guide, and a new set of questions, the Eating disorder Screen for Primary care (ESP), using the Questionnaire for Eating Disorders Diagnosis as the independent standard, in 104 consecutive patients from a primary care practice and 129 university students. Twelve percent of the combined population had an eating disorder. One or no abnormal responses to the ESP ruled out an eating disorder (likelihood ratio [LR] 0.0), whereas 3 or more abnormal responses ruled one in (LR 11). The SCOFF questions were less sensitive than predicted (1 or no abnormal responses, LR 0.25), but were as effective at ruling in an eating disorder (3 or more abnormal responses, LR 11). PMID- 12534765 TI - Adverse drug event monitoring at the Food and Drug Administration. AB - The Food and Drug Administration (FDA) is responsible not only for approving drugs but also for monitoring their safety after they reach the market. The complete adverse event profile of a drug is not known at the time of approval because of the small sample size, short duration, and limited generalizability of pre-approval clinical trials. This report describes the FDA's postmarketing surveillance system, to which many clinicians submit reports of adverse drug events encountered while treating their patients. Despite its limitations, the spontaneous reporting system is an extremely valuable mechanism by which hazards with drugs that were not observed or recognized at the time of approval are identified. Physicians are strongly encouraged to submit reports of adverse outcomes with suspect drugs to the FDA, and their reports make a difference. The FDA is strengthening its postmarketing surveillance with access to new data sources that have the potential to further improve the identification, quantification, and subsequent management of drug risk. PMID- 12534766 TI - Measuring errors and adverse events in health care. AB - In this paper, we identify 8 methods used to measure errors and adverse events in health care and discuss their strengths and weaknesses. We focus on the reliability and validity of each, as well as the ability to detect latent errors (or system errors) versus active errors and adverse events. We propose a general framework to help health care providers, researchers, and administrators choose the most appropriate methods to meet their patient safety measurement goals. PMID- 12534767 TI - Integrating medical care and addiction treatment. PMID- 12534768 TI - Barriers to postfracture osteoporosis care in postmenopausal women. PMID- 12534769 TI - Remedies needed to address the pathology in reporting adverse reactions and Food and Drug Administration use of reports. PMID- 12534770 TI - History of chronic obstructive pulmonary disease (COPD), wheezes, and forced expiratory time (FET)--in evaluating test measures for COPD and for assessing time intervals for performing bedside maneuvers. PMID- 12534771 TI - The likelihood ratio as one of--if not the most important--operating characteristic of a diagnostic test. PMID- 12534774 TI - The management of portal hypertension: controversies. PMID- 12534775 TI - The role of aflatoxins and hepatitis viruses in the etiopathogenesis of hepatocellular carcinoma: A basis for primary prevention in Guinea-Conakry, West Africa. AB - Aflatoxins and hepatitis B virus (HBV) are major risk factors for hepatocellular carcinoma (HCC) in South-east Asia and Africa, parts of the world where this cancer is most prevalent. Exposure to both factors is endemic, occurring from early in life. There is evidence from both epidemiological studies and animal models that the two factors can act synergistically to increase the risk of HCC, but the underlying cellular and molecular mechanisms of interaction are as yet undefined. One possibility suggested by studies in HBV transgenic mice is that chronic liver injury alters the expression of carcinogen metabolizing enzymes, thus modulating the level of binding of aflatoxin to DNA. Primary prevention of HCC in high incidence areas of the world should primarily be focused on provision of the safe, effective vaccine against HBV. However, measures to reduce the high levels of aflatoxin exposure, where chronic HBV infection is currently epidemic, would also significantly contribute to reducing HCC incidence. In Guinea-Conakry, West Africa, surveys of HBV infection and aflatoxin exposure have established baseline data for the implementation of a community-based intervention study. This study will evaluate the effectiveness of improving the post-harvest processing and storage of the groundnut crop, a major source of aflatoxins, using aflatoxin-albumin adducts as the outcome measurement. PMID- 12534776 TI - Overview: past and future of immunologic intervention in the pathogenesis, prophylaxis and therapeusis of hepatitis B. PMID- 12534777 TI - Public health measures in the control of viral hepatitis: a World Health Organization perspective for the next millennium. PMID- 12534778 TI - Animal models for the study of HBV infection and the evaluation of new anti-HBV strategies. AB - BACKGROUND: Our aim was to evaluate the anti-HBV activity of a novel L-nucleoside analog, 2',3'-dideoxy-2',3'-didehydro-beta-L-5-fluorocytidine (beta-L-Fd4C), in study models of HBV infection. METHOD: Its mechanism of action was evaluated on the in vitro expressed duck HBV (DHBV) reverse transcriptase and in primary hepatocyte cultures of duck and human origin. The capacity of antiviral therapy to clear viral infection was analyzed in vivo in the duck and woodchuck models. RESULTS: beta-L-Fd4C-TP exhibited a more potent inhibitory effect on the RT activity of the DHBV polymerase than other cytidine analogs (lamivudine-TP, ddC TP, beta-L-FddC-TP). In primary duck hepatocyte cultures, beta-L-Fd4C exhibited a long-lasting inhibitory effect on viral DNA synthesis but could not clear viral cccDNA. In vivo treatment with beta-L-Fd4C in infected ducklings and woodchucks, induced a greater suppression of viremia and intrahepatic viral DNA synthesis than with lamivudine. However, covalently closed circular DNA persistence explained the relapse of viral replication after treatment withdrawal. Viral spread was strongly reduced in the case of early therapeutical intervention, but the number of infected cells did not decline when therapy was started during chronic infection. Liver histology analysis showed a decrease in the inflammatory activity of chronic hepatitis while no ultrastructural modification of liver cells was observed in electron microscopy studies. Furthermore, in human primary hepatocyte cultures, beta-L-Fd4C induced a significant inhibition of HBV DNA synthesis. CONCLUSION: beta-L-Fd4C is a potent inhibitor of hepadnavirus RT and inhibits viral DNA synthesis in hepatocytes both in vitro and in vivo. These experimental studies allowed as to show that beta-L-Fd4C is a promising anti-HBV agent. Combination therapy should be evaluated to eradicate viral infection. PMID- 12534779 TI - Hepatitis viruses: a pandora's box? AB - The term hepatitis virus is reserved for those viruses that are predominantly hepatotropic, although several new agents have been assigned to this category in the absence of hepatotropism and clinical disease. The hepatitis viruses can be broadly divided into those transmitted via the fecal-oral route, and those by blood, blood products and body fluids. Hepatitis A (picornaviridae), hepatitis B (hepadnaviridae) and hepatitis C (flaviviridae) represent the major public health problems. The epidemiology of hepatitis A virus (HAV) and hepatitis B virus (HBV) is changing in response to vaccination. In the case of HAV, older age groups are now deemed at risk, particularly of fulminant hepatitis if exposed over the age of 50. Chronic hepatitis B in some regions is now predominantly of the so-called precore mutant type where high levels of HBV replication persist in the presence of anti-hepatitis B virus (HBe) antibodies. The HBV vaccination is among the most cost-effective health care measures. The epidemiological significance of mutations found increasingly in the HBV S gene isolated from vaccinated children is unclear. Evidence that hepatitis G and TT virus are significant causes of hepatitis is lacking. Of interest, however, is the finding that the related GBV-B agent of monkeys may be a model for developing new antiviral agents against HCV. Animal models of hepatitis infections are providing new insights into the pathogenesis of hepatitis in humans. Indeed it is possible that hepatitis E is primarily an agent of pigs and other domesticated livestock. Intriguingly, the new TT virus shares many properties with the circoviruses, significant pathogens of chickens and pigs. The challenge in the next decade will be to assess the significance of these new agents in terms of public health and resources. Value judgements will have to be made in assessing the risks associated with blood containing trace amounts of these adventitious agents. PMID- 12534780 TI - Hepatitis C virus: clades and properties. AB - Hepatitis C virus (HCV) is a member of the virus family Flaviviridae. At present HCV is classified into a discrete hepacivirus genus and is represented by six clades according to genome sequencing. Each clade is further divisible into subtypes, which may prove important for the study of clinical differences and epidemiological studies. Limited homology also exists with hepatitis G/GB viruses, despite the fact that the hepatotropic nature of the latter agents remains contentious. The variability amongst the six HCV clades is less than that observed between the four serotypes of dengue, suggesting that each clade may represent a distinct virus were tests such as plaque neutralization to become available for delineating HCV isolates. The distribution worldwide varies, with Clades 1 and 2 predominating in most regions-an important consideration for the development of any vaccine. In addition, the clade distribution among cohorts may vary according to age. Point source outbreaks of HCV, for example in large numbers of women inadvertently infected with HCV-contaminated anti-D globulin, offers an opportunity to study the evolution of HCV genotypes over several decades. Parallel studies in chimpanzees have shown that the hypervariable region of E2 may play a role in HCV immunity, with quasispecies rapidly replacing the predominant subtype as immunity develops to the initiating virus strain. There is some evidence that an IFN-sensitive motif exists in the NS5 gene which may have some predictive value in determining the likely outcome of IFN treatment. A database is available for all HCV sequences, together with information about their properties and guidance for the evaluation of new isolates (http://s2as02.genes.nig.ac.jp). PMID- 12534781 TI - Emerging therapies of hepatitis B and C. PMID- 12534782 TI - Pathophysiology of portal hypertension. PMID- 12534783 TI - Immunological tolerance: the liver effect. PMID- 12534784 TI - Molecular characterization of hepatitis B virus surface antigen mutants in Singapore patients with hepatocellular carcinoma and hepatitis B virus carriers negative for HBsAg but positive for anti-HBs and anti-HBc. AB - BACKGROUND AND AIMS: Mutations on the a-determinant of hepatitis B virus surface antigen (HBsAg), capable of escaping detection and vaccination, are identified in HBsAg-positive/anti-HBs-positive vaccinated infants. We studied the prevalence of these mutants in HBsAg-negative/anti-HBc-positive chronic HBV carriers and patients with hepatocellular carcinoma (HCC). METHODS: DNA sequence coding for the antigenic a-determinant of HBsAg was amplified from either HCC genomic DNA or serum samples of the selected patients and sequenced. The replicative mutant genomes were reconstituted in vitro and their reactivity to commercial kits measured. RESULTS: Mutations within and/or outside the a-determinant were identified in patients seronegative for HBsAg. They were then reconstituted in vitro and transiently transfected into HepG2 cells. Culture medium containing secreted HBV viral particles was collected and assayed for their binding to commercial kits. Drastic decrease of reactivity to these kits was seen with most of the identified mutations, including those located outside the a-determinant. CONCLUSION: The existence of a more complex antigenic structure of HBsAg is indicated by the decreased reactivity to detection of mutations, some of which are outside the a-determinant, escape vaccination and may persist in seronegative patients. The high proportion of HBsAg mutants that are integrated in HCC genomes suggests a role of these mutants in hepatocarcinogenesis, possibly leading to mutant HBV-related HCC. PMID- 12534785 TI - Hepatitis B virus mutants: an overview. PMID- 12534786 TI - Prevention of human hepatocellular carcinoma with natural lymphoblastoid alpha interferon. AB - Hepatitis B surface antigen (HBsAg) positive hepatocellular carcinoma (HCC) patients have a high risk of recurrence within 5 years of hepatic resection and suitable long-term maintenance therapy is required. An open-ended controlled trial on 30 HBsAg positive Child Pugh's grade A cirrhotic patients was started in September 1986 using Wellferon (natural lymphoblastoid alpha interferon). Ten patients had standard adriamycin and mitomycin C monthly for 6 months, then discontinued when the course was completed. Twenty other patients had this regime, plus the addition of Wellferon given at 3 MU i.m. daily for 10 days. No HCC recurrence was observed in the interferon group. Followed to 14 years, 11 of the interferon-treated patients are still well and HCC free, whereas none of the chemotherapy patients are alive. High incidence of recurrence occurred in those who discontinued interferon (IFN), reduced the dose, or took the scheduled dose, but at longer intervals of more than 3 months. Wellferon, a natural alpha-IFN has potent inhibitory effects and reduces the risk of HCC development. It is a suitable agent for long-term maintenance prophylactic therapy for high-risk resected HBsAg positive HCC patients. PMID- 12534787 TI - The discovery of the hepatitis B virus and the invention of the vaccine: a scientific memoir. PMID- 12534788 TI - Liver disease history in India. PMID- 12534789 TI - Past hepatitis studies in Japan. PMID- 12534790 TI - History of hepatitis and liver disease in Indonesia. PMID- 12534791 TI - Memoir on hepatitis and liver disease history in Hong Kong. PMID- 12534797 TI - The spectrum of cutaneous lesions in rheumatoid arthritis: a clinical and pathological study of 43 patients. AB - INTRODUCTION: Rheumatoid arthritis (RA) is an idiopathic arthropathy syndrome that has a propensity to affect the small joints of the hands and feet with extra articular manifestations comprising skin lesions, neuropathy, pericarditis, pleuritis, interstitial pulmonary fibrosis and a systemic polyarteritis nodosa (PAN)-like vasculitic syndrome. The most widely recognized skin lesion is the rheumatoid nodule. Other skin manifestations are poorly defined. MATERIALS AND METHODS: Using a natural language search of the authors' outpatient dermatopathology databases, skin biopsies from 43 patients with RA were selected for retrospective analysis in an attempt to define the dermatopathological spectrum of RA and its clinical correlates. RESULTS: The biopsies were categorized by the dominant histologic pattern, recognizing that in most cases there were additional minor reaction patterns. Palisading and/or diffuse interstitial granulomatous inflammation was the dominant pattern seen in 21 patients; the lesions included nodules, plaques and papules with a predilection to involve skin over joints. Besides interstitial histiocytic infiltrates and variable collagen necrobiosis, these cases also showed interstitial neutrophilia, vasculitis and pauci-inflammatory vascular thrombosis. The dominant morphology in 11 other patients was vasculopathic in nature: pauci-inflammatory vascular thrombosis, glomeruloid neovascularization, a neutrophilic vasculitis of pustular, folliculocentric, leukocytoclastic or benign cutaneous PAN types, granulomatous vasculitis, and lymphocytic vasculitis and finally occlusive intravascular histiocytic foci for which the designation of "RA-associated intravascular histiocytopathy" is proposed. Rheumatoid factor (RF) positivity and active arthritis were common in this group, with anti-Ro and anticardiolipin antibodies being co-factors contributing to vascular injury in some cases. Immunofluorescent testing in three patients revealed dominant vascular IgA deposition. In nine patients, the main pattern was one of neutrophilic dermal and/or subcuticular infiltrates manifested clinically as urticarial plaques, pyoderma gangrenosum and panniculitis. CONCLUSIONS: The cutaneous manifestations of RA are varied and encompass a number of entities, some of which define the dominant clinical features, such as the rheumatoid papule or subcutaneous cords, while others allude to the histopathology, i.e. rheumatoid neutrophilic dermatosis. We propose a more simplified classification scheme using the adjectival modifiers of "rheumatoid-associated" and then further categorizing the lesion according to the dominant reaction pattern. Three principal reaction patterns are recognized, namely extravascular palisading granulomatous inflammation, interstitial and/or subcuticular neutrophilia and active vasculopathy encompassing lymphocyte-dominant, neutrophil-rich and granulomatous vasculitis. In most cases, an overlap of the three reaction patterns is seen. Co factors for the vascular injury that we believe are integral to the skin lesions of RA include RF, anti-endothelial antibodies of IgA class, anti-Ro and anticardiolipin antibodies. PMID- 12534798 TI - p63 expression in normal human epidermis and epidermal appendages and their tumors. AB - BACKGROUND: p63, a member of the p53 gene family, is expressed in basal cells of several different organs. METHODS: The immunoreactivity of p63 was examined in normal human epidermis and epidermal appendages and their tumors, and compared with proliferative activity as evaluated by Ki-67. RESULTS: In normal skin, p63 expression was seen in basal/suprabasal cells of the epidermis, outer root sheath and hair matrix cells of the hair follicle, seboblast situated in the outermost layer of sebaceous glands, and outer layer cells of the ductal portion and myoepithelial cells of the secretory portion of the sweat glands. p63 expression was confined to the cells forming a continuous basal rim along the normal epithelial structure. In tumors, p63 expression resembled that in normal tissue in that tumor components originating from p63-positive cells were constantly positive for p63. In normal and tumor tissues, not all p63-positive cells were positive for Ki-67. CONCLUSIONS: p63 expression may be a marker of basal/progenitor cells in tumors of epidermis and epidermal appendages, and may be a diagnostic marker of these tumors. PMID- 12534799 TI - Expression of syndecan-1 is a sensitive marker for cutaneous plasmacytoma. AB - BACKGROUND: Cutaneous plasmacytoma is a well-recognized, yet infrequent, occurrence in multiple myeloma (MM). There are limitations in the morphologic assessment, and as such, the diagnosis presents some difficulty, particularly with the plasmablastic type. METHODS: Pathology reports of 2357 patients with a diagnosis of MM were reviewed. Twenty patients yielded a total of 25 plasmacytomas, 10 of which were analyzed for syndecan-1 immunoreactivity. Bartl grade of bone marrow and cutaneous plasmacytoma was compared and immunoglobulin secretory status of the patients was assessed. RESULTS: The incidence of cutaneous plasmacytoma was found to be 1 in 118 patients with MM. Immunoglobulin secretion was found to be predominantly IgG. There was a trend for the plasmacytoma Bartl grade to be equal to or greater than that of the corresponding bone marrow Bartl grade, suggesting a more aggressive phenotype in the metastatic lesion. CONCLUSION: Syndecan-1 was found to be a sensitive marker for plasmacytomas, independent of cytologic differentiation. PMID- 12534800 TI - Fatty acid synthase expression in melanoma. AB - BACKGROUND: Fatty acid synthase (FAS), the key enzyme responsible for the synthesis of fatty acids, is weakly expressed in some normal human tissues. Recently, FAS has been demonstrated to be overexpressed in many non-neoplastic highly proliferative lesions and in aggressive carcinomas with poor outcome, including colon, breast and ovary carcinomas. METHODS: In order to evaluate the prognostic significance of FAS in human melanoma, we analysed by means of immunohistochemistry, using a monoclonal anti-FAS antibody, 77 primary melanomas and 30 nodal and cutaneous metastasis. Thirty nevi (15 dermal and 15 junctional nevi) were used as controls. All patients were followed-up for 5 years. RESULTS: Thirty-four melanomas expressed strong FAS immunostaining; the remaining 43 cases showed weak expression or were negative. All cutaneous and nodal metastasis were strongly positive. All patients with metastases deceased during the follow up period. Control specimens expressed weak staining. None of these patients developed recurrence. Statistical analysis revealed significant association of FAS expression with Breslow thickness (p = 0.012). The intensity of FAS immunostaining was also predictive of prognosis (p = 0.049). CONCLUSIONS: FAS is a reliable prognostic marker in human melanomas. FAS predictive strength is increased when associated with Breslow thickness. The observation of FAS in human melanomas may stratify patients for stricter follow-ups and suggest different therapeutic approaches. PMID- 12534801 TI - Multiple follicular cysts, infundibular type with vellus hairs and solar elastosis of the ears: a new dermatoheliosis? AB - BACKGROUND: Vellus hair cyst is an uncommon developmental abnormality of the vellus follicle histologically defined as a stratified squamous epithelial-lined cyst containing one or more vellus hairs. METHODS: Herein, we report three patients with a heretofore described clinicopathologic condition consisting of multiple contiguous cysts of the ear helices showing solar elastosis and multiple cysts containing vellus hairs, microscopically. CONCLUSION: Given the anatomic location, history of excessive ultraviolet exposure and pathologic alterations, we surmise that this is a condition related to Favre-Racouchot syndrome, which is predisposed by excessive ultraviolet light exposure. PMID- 12534803 TI - Localized amyloidosis of the glans penis: a case report and literature review. AB - BACKGROUND: Primary localized cutaneous amyloidosis is an uncommon lesion with a varied pathogenesis. METHODS: We report the case of a 67-year-old-male discovered to have a localized amyloid lesion of the glans penis. RESULTS: Biopsy of the lesion revealed dermal deposits of amorphous eosinophilic material which stained positive with Congo red and amyloid P protein. Additional stains, including kappa and lambda light chains, amyloid A, and transthyretin, were negative. The lesion has remained asymptomatic, with no evidence of systemic disease identified, and no further treatment has been necessary. CONCLUSIONS: This is the sixth reported case of localized amyloidosis of the glans penis. Based on the clinical behavior and pathologic characteristics, this type of lesion is best classified as primary localized cutaneous amyloidosis, in the same family as the macular/lichenoid type lesions. PMID- 12534802 TI - Syringocystadenoma papilliferum contiguous to a verrucous cyst. AB - BACKGROUND: Syringocystadenoma papilliferum (SCAP) and verrucous cyst are two uncommon benign tumors. The simultaneous occurrence of the two lesions has not been reported before. METHODS: We report four cases of the simultaneous occurrence of the two rare lesions as so-called collision lesions with a review of the literature. CONCLUSION: The relationship of SCAP with viral infection needs further investigation. PMID- 12534806 TI - An antifungal compound produced by Bacillus subtilis YM 10-20 inhibits germination of Penicillium roqueforti conidiospores. AB - AIMS: To identify and characterize an antifungal compound produced by Bacillus subtilis YM 10-20 which prevents spore germination of Penicillium roqueforti. METHODS AND RESULTS: The antifungal compound was isolated by acid precipitation with HCl. This compound inhibited fungal germination and growth. Identification by HPLC and mass spectrometry analysis showed high similarity to iturin A. Permeabilization and morphological changes in P. roqueforti conidia in the presence of the inhibitor were revealed by fluorescence staining and SEM, respectively. CONCLUSOINS: The iturin-like compound produced by B. subtilis YM 10 20 permeabilizes fungal spores and blocks germination. SIGNIFICANCE AND IMPACT OF THE STUDY: Fluorescence staining in combination with flow cytometry and scanning electron microscopy are efficient tools for assessing the action of antifungal compounds against spores. Iturin-like compounds may permeabilize fungal spores and inhibit their germination. PMID- 12534807 TI - Influence of phenolic acids on growth and inactivation of Oenococcus oeni and Lactobacillus hilgardii. AB - AIMS: To determine the effect of several wine-associated, phenolic acids on the growth and viability of strains of Oenococcus oeni and Lactobacillus hilgardii. METHODS AND RESULTS: Growth was monitored in ethanol-containing medium supplemented with varying concentrations of hydroxybenzoic acids (p hydroxybenzoic, protocatechuic, gallic, vanillic and syringic acids) and hydroxycinnamic acids (p-coumaric, caffeic and ferulic acids). Progressive inactivation was monitored in ethanol-containing phosphate buffer supplemented in a similar manner to the growth experiments. Hydroxycinnamic acids proved to be more inhibitory to the growth of O. oeni than hydroxybenzoic acids. On the other hand, some acids showed a beneficial effect on growth of Lact. hilgardii. p Coumaric acid showed the strongest inhibitory effect on growth and survival of both bacteria. CONCLUSIONS: Most phenolic acids had a negative effect on growth of O. oeni, for Lact. hilgardii this effect was only noted for p-coumaric acid. Generally, O. oeni was more sensitive to phenolic acid inactivation than Lact. hilgardii. SIGNIFICANCE AND IMPACT OF THE STUDY: Eight wine-derived, phenolic acids were compared for their effects on wine lactic acid bacteria. Results indicate that phenolic acids have the capacity to influence growth and survival parameters. The differences found between phenolic compounds could be related to their different chemical structures. PMID- 12534808 TI - UDP-galactose 4-epimerase: a key enzyme in exopolysaccharide formation by Lactobacillus casei CRL 87 in controlled pH batch cultures. AB - AIMS: To evaluate the relationship between exopolysaccharide (EPS) production and the sugar nucleotide biosynthetic enzymes in Lactobacillus casei CRL 87 under optimum growth conditions for polymer formation: controlled pH on galactose or glucose. Studies with an EPS mutant were carried out to determine the key enzymes in EPS synthesis under the above culture conditions. METHODS AND RESULTS: EPS concentration was estimated by the phenol/sulphuric acid method, while the activities of the biosynthetic enzymes were determined spectrophotometrically by measuring the formation or disappearance of NAD(P)H at 340 nm. An environmental pH of 5.0, using galactose as carbon source, markedly improved not only polymer production and yield but also, cell growth and lactic acid production. Analysis of the activities of the EPS precursor-forming enzymes revealed that polysaccharide synthesis was correlated with uridine-diphosphate (UDP)-glucose pyrophosphorylase and UDP-galactose 4-epimerase under these growth conditions. CONCLUSIONS: EPS synthesis by Lact. casei CRL 87 was considerably improved at a controlled pH of 5.0 with galactose as carbon source, and was correlated with the activity of UDP-glucose pyrophosphorylase and UDP-galactose 4-epimerase. The results obtained with the wild-type and EPS- strains suggest that UDP-galactose 4 epimerase plays an essential role in EPS formation. SIGNIFICANCE AND IMPACT OF THE STUDY: Unravelling the key enzymes involved in EPS biosynthesis under optimum culture conditions for polymer production provides important information for the design of strategies, via genetic engineering, to enhance polysaccharide formation. PMID- 12534809 TI - The effect of acid shock on sporulating Bacillus subtilis cells. AB - AIMS: To study the effect of acid shock in sporulation on the production of acid shock proteins, and on the heat resistance and germination characteristics of the spores formed subsequently. METHODS AND RESULTS: Bacillus subtilis wild-type (SASP-alpha+beta+) and mutant (SASP-alpha-beta-) cells in 2 x SG medium at 30 degrees C were acid-shocked with HCl (pH 4, 4.3, 5 and 6 against a control pH of 6.2) for 30 min, 1 h into sporulation. The D85-value of B. subtilis wild-type (but not mutant) spores formed from sporulating cells acid-shocked at pH 5 increased from 46.5 min to 78.8 min, and there was also an increase in the resistance of wild-type acid-shocked spores at both 90 degrees C and 95 degrees C. ALA- or AGFK-initiated germination of pH 5-shocked spores was the same as that of non-acid-shocked spores. Two-dimensional gel electrophoresis showed only one novel acid-shock protein, identified as a vegetative catalase 1 (KatA), which appeared 30 min after acid shock but was lost later in sporulation. CONCLUSIONS: Acid shock at pH 5 increased the heat resistance of spores subsequently formed in B. subtilis wild type. The catalase, KatA, was induced by acid shock early in sporulation, but since it was degraded later in sporulation, it appears to act to increase heat resistance by altering spore structure. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first proteomic study of acid shock in sporulating B. subtilis cells. The increasing spore heat resistance produced by acid shock may have significance for the heat resistance of spores formed in the food industry. PMID- 12534810 TI - Investigation of Salmonella contamination and disinfection in farm egg-packing plants. AB - AIMS: As part of a field-based study of the distribution and persistence of Salmonella infection on commercial egg-laying farms, sampling was carried out on one or more occasions in egg-packing areas of 12 farms infected with Salm. Enteritidis. METHODS AND RESULTS: Salmonellas were isolated by cultural methods. Contamination was common, with Salmonella being found in 23.1% of floor swab samples, 30.8% of grading tables, 23.1% of conveyor belts or rollers and 23.8% of candlers. Four farms were sampled after cleaning and disinfection of packing plants had been carried out on the previous day, and residual contamination was found on 6.9% of samples from grading tables, 16.0% holding/sorting tables, 12.6% of conveyors or rollers, 16.7% of vacuum egg lifters, 21.4% of floor surfaces and 5.0% of egg store floor surfaces. Sterilized eggs passed through five farm packing plants showed a contamination rate of at least 16/5,948 (0.3%) egg passages. CONCLUSIONS: It is apparent that contamination in egg-packing plants may be a significant contributory factor to external contamination of shell eggs, and improved methods of cleaning and disinfecting egg-handling equipment are required. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of Salmonella contamination in egg-packing plants presents a contamination hazard for eggs from Salmonella-free flocks. Samples from equipment in the packing plant could also be used for screening for detection of Salmonella in the throughout of the plant. PMID- 12534811 TI - Employment of broad-range 16S rRNA PCR to detect aetiological agents of infection from clinical specimens in patients with acute meningitis--rapid separation of 16S rRNA PCR amplicons without the need for cloning. AB - AIMS: The aim of this study was to develop a polyacrylamide gel electrophoresis (PAGE) method for the rapid separation of 16S rRNA PCR amplicons from aetiological agents of acute meningitis. METHODS AND RESULTS: Blood samples from 40 patients with suspected acute meningococcal meningitis were examined for the presence of causal agents, including Neisseria meningitidis employing two methods: (i) broad-range 16S rRNA PCR in conjunction with PAGE and automated sequencing and (ii) species-specific PCR employing ABI TaqMan technology for N. meningitidis. Analysis of clinical specimens employing 16S rRNA PCR yielded 33/40 (82.5%) positive for the presence of bacterial DNA. Species-specific PCR yielded 30/40 (75%) clinical specimens positive for N. meningitidis. Prior to separation by PAGE, only 6/33 (18.2%) amplicons were able to be identified by sequence analysis, the remaining amplicons (n=27) did not yield an identification due to the presence of mixed 16S rRNA PCR amplicons. Following separation, amplicons were re-amplified and sequenced, yielding 24/27 (88.9%) positive for N. meningitidis and three specimens positive for Acinetobacter sp., Staphylococcus aureus and Streptococcus pneumoniae. One specimen was positive for both N. meningitidis and Streptococcus spp. and another specimen was positive for N. meningitidis and Pseudomonas sp., by broad-range PCR. Seven clinical specimens were negative for N. meningitidis and other eubacteria using both detection techniques. CONCLUSIONS: Clinical specimens including blood and cerebrospinal fluid from patients with suspected acute bacterial meningitis, may become contaminated with commensal skin flora, resulting in difficulties in downstream sequencing of pathogen plus contaminant DNA. This study allows for the rapid separation of amplified pathogen from contaminant DNA. SIGNIFICANCE AND IMPACT OF STUDY: This study demonstrated the usefulness of the rapid separation of multiple 16S rRNA PCR amplicons using a combination of PAGE and automated sequencing, without the need of cloning. Adoption of this technique is therefore proposed when trying to rapidly identify pathogens in clinical specimens employing broad range 16S rRNA PCR. PMID- 12534812 TI - Effects of the antibiotic ionophores monensin, lasalocid, laidlomycin propionate and bambermycin on Salmonella and E. coli O157:H7 in vitro. AB - AIMS: To examine the effects of ionophores on Salmonella and Escherichia coli O157:H7 in pure and mixed ruminal fluid cultures. METHODS AND RESULTS: Four Salmonella serotypes (Dublin, Derby, Typhimurium, and Enteriditis) and two strains of E. coli O157:H7 (ATCC 43895 and FDIU 6058) were cultured in the presence of varying concentrations of ionophores (monensin, lasalocid, laidlomycin propionate, and bambermycin) in pure and mixed ruminal fluid cultures. Bacterial growth rates in pure culture were not affected (P > 0.10) by ionophores at concentrations up to 10 times the approximate rumen ionophore concentration under normal feeding regimens. Likewise, ionophores had no effect (P > 0.10) on Salmonella or E. coli CFU plated from 24-h ruminal fluid incubations. Ionophore treatment decreased (P < 0.01) the acetate : propionate ratio in ruminal fluid cultures as expected. CONCLUSIONS: Ionophores had no effect on the foodborne pathogens Salmonella and E. coli O157:H7 in vitro. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that ionophore feeding would have little or no effect on Salmonella or E. coli populations in the ruminant. PMID- 12534813 TI - Isolation and biochemical characterisation of enterocins produced by enterococci from different sources. AB - AIMS: Comparison of enterocins produced by six Enterococcus faecium strains and one Ent. faecalis strain isolated from different origin with regard to their microbiological and biochemical characteristics in view of their technological potential and practical use. METHODS AND RESULTS: The seven enterococci were sensitive to the glycopeptide antibiotics vancomycin and teicoplanin and did not show haemolytic activity. The absence of the glycopeptide-resistant genotypes and the genes involved in the production of the lantibiotic cytolysin was confirmed by PCR. The enterocins were active towards Listeria innocua and other lactic acid bacteria. Their temperature stability was dependent on the pH and their activity was higher at acidic pH. A bactericidal and bacteriolytic effect was shown. PCR analyses revealed that the gene of enterocin A was present in the genome of Ent. faecium CCM 4231, Ent. faecium 306 I.2.20 and Ent. faecalis Y; both enterocin A and B genes were present in the genome of Ent. faecium LMG 11423T, Ent. faecium RZS C5 and Ent. faecium RZS C13. Enterocin P was detected in the genome of Ent. faecium RZS C5 and Ent. faecium RZS C13. No signal was found for Ent. faecium SF 68. Enterocins from Ent. faecium RZS C5, Ent. faecium RZS C13 and Ent. faecium SF 68 were purified to homogeneity. CONCLUSIONS: Ent. faecium RZS C5 and Ent. faecium RZS C13 produced an enterocin with a molecular mass of 5460 and 5477 Da, respectively, which was in the range of that of enterocin B. The amino acid sequence analysis of the enterocin from Ent. faecium RZS C13 revealed 24 N terminal residues, which were identical to those of enterocin B. The enterocin from Ent. faecium SF 68 had a molecular mass of 4488 Da, which did not correspond to any enterocin known so far. SIGNIFICANCE AND IMPACT OF THE STUDY: The number of characterized enterocins is increasing. As this type of work is tedious and time-consuming, it may be interesting to include PCR as a first step to know if the Enterococcus strain in study produces either a known or a new enterocin. Also, it is important to check the absence of cytolysin and resistance to vancomycin for a further application of the Enterococcus strain in food or health applications. PMID- 12534814 TI - Degradation of straight-chain aliphatic and high-molecular-weight polycyclic aromatic hydrocarbons by a strain of Mycobacterium austroafricanum. AB - AIMS: Our goal was to characterize a newly isolated strain of Mycobacterium austroafricanum, obtained from manufactured gas plant (MGP) site soil and designated GTI-23, with respect to its ability to degrade polycyclic aromatic hydrocarbons (PAHs). METHODS AND RESULTS: GTI-23 is capable of growth on phenanthrene, fluoranthene, or pyrene as a sole source of carbon and energy; it also extensively mineralizes the latter two in liquid culture and is capable of extensive degradation of fluorene and benzo[a]pyrene, although this does not lead in either of these cases to mineralization. Supplementation of benzo[a]pyrene containing cultures with phenanthrene had no significant effect on benzo[a]pyrene degradation; however, this process was substantially inhibited by the addition of pyrene. Extensive and rapid mineralization of pyrene by GTI-23 was also observed in pyrene-amended soil. CONCLUSIONS: Strain GTI-23 shows considerable ability to mineralize a range of polycyclic aromatic hydrocarbons, both in liquid and soil environments. In this regard, GTI-23 differs markedly from the type strain of Myco. austroafricanum (ATCC 33464); the latter isolate displayed no (or very limited) mineralization of any tested PAH (phenanthrene, fluoranthene or pyrene). When grown in liquid culture, GTI-23 was also found to be capable of growing on and mineralizing two aliphatic hydrocarbons (dodecane and hexadecane). SIGNIFICANCE AND IMPACT OF THE STUDY: These findings indicate that this isolate of Myco. austroafricanum may be useful for bioremediation of soils contaminated with complex mixtures of aromatic and aliphatic hydrocarbons. PMID- 12534815 TI - Activity and mechanisms of action of selected biocidal agents on Gram-positive and -negative bacteria. AB - AIMS: This study investigates the antimicrobial activity and mode of action of two natural products, eugenol and thymol, a commonly utilized biostatic agent, triclocarban (TCC), and two surfactants, didecyldimethylammonium chloride (DDDMAC) and C10-C16 alkyldimethyl amine N-oxides (ADMAO). METHODS AND RESULTS: Methods used included: determination of minimum inhibitory concentrations (MICs), lethal effect studies with suspension tests and the investigation of sub-MIC concentrations on growth of E. coli, Staph. aureus and Ps. aeruginosa using a Bioscreen microbiological analyser. Leakage of intracellular constituents and the effects of potentiating agents were also investigated. Only DDDMAC was bactericidal against all of the organisms tested. Eugenol, thymol and ADMAO showed bacteriostatic and bactericidal activity, but not against Ps. aeruginosa. TCC was only bacteristatic against Staph. aureus, but like the other agents, it did affect the growth of the other organisms in the Bioscreen experiments. All of the antimicrobial agents tested were potentiated by the permeabilizers to some extent and leakage of potassium was seen with all of the agents except TCC. CONCLUSIONS: DDDMAC was bactericidal against all organisms tested and all compounds had some bacteriostatic action. Low level static effects on bacterial growth were seen with sub-MIC concentrations. Membrane damage may account for at least part of the mode of action of thymol, eugenol, DDDMAC and ADMAO. SIGNIFICANCE AND IMPACT OF THE STUDY: The ingredients evaluated demonstrated a range of bactericidal and bacteriostatic properties against the Gram-negative and -positive organisms evaluated and the membrane (leakage of intracellular components) was implicated in the mode of action for most (except TCC). Sub-MIC levels of all ingredients did induce subtle effects on the organisms which impacted bacterial growth, even for those which had no true inhibitory effects. PMID- 12534816 TI - Response of bacterial community during bioremediation of an oil-polluted soil. AB - AIM: To study the response of the bacterial community to bioremediation of a soil with an aged contamination of crude oil. METHODS AND RESULTS: The bacterial community in laboratory soil columns during a 72-day biostimulation treatment was followed by analysing the number of total cultivable hydrocarbon-degrading bacteria, soil respiratory activity and the 16S-23S rDNA internal transcribed spacer homoduplex heteroduplex polymorphisms (ITS-HHP) of total soil bacterial DNA. ITS-HHP permits an estimate of both length and sequence polymorphism in a 16S-23S rDNA spacer population, using to advantage the homoduplex and heteroduplex fragments that are generated during PCR. The treatment, made by air sparging and biostimulation with a mineral nutrient and surfactant solution, resulted in a 39.5% decrease of the total hydrocarbon content. Within 4 days of treatment onset the bacterial community underwent a first phase of activation that led to a substantial increase in the observable diversity. Subsequently, after a 12-day period of stability, another activation phase was observed with further shifts of the community structure and an increase in the abundance and diversity of catechol-2,3-dioxygenase (C23O) genes. CONCLUSIONS: The overall data suggest an important contribution of uncultivable bacteria to the soil bioremediation, since, during the second activation phase, the increases of the respiratory activity, bacterial diversity and C23O gene abundance and diversity were not accompanied by a corresponding increase of the cultivable bacteria number. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that successive phases of activation of bacterial populations occur during a bioremediation treatment of oil-polluted soil. PMID- 12534817 TI - Mechanisms underlying the toxicity of lactone aroma compounds towards the producing yeast cells. AB - AIMS: To study the fundamental mechanisms of toxicity of the fruity aroma compound gamma-decalactone, that lead to alterations in cell viability during its biotechnological production by yeast cells; Yarrowia lipolytica that is able to produce high amounts of this metabolite was used here as a model. METHODS AND RESULTS: Lactone concentrations above 150 mg l-1 inhibited cell growth, depolarized the living cells and increased membrane fluidity. Infrared spectroscopic measurements revealed that the introduction of the lactone into model phospholipid bilayers, decreased the phase transition temperature. Moreover, the H+-ATPase activity in membrane preparations was strongly affected by the presence of the lactone. On the other hand, only a slight decrease in the intracellular pH occurred. CONCLUSIONS: We propose that the toxic effects of gamma-decalactone on yeast may be initially linked to a strong interaction of the compound with cell membrane lipids and components. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings may enable the elaboration of strategies to improve yeast cell viability during the process of lactones bioproduction. PMID- 12534818 TI - Degradation of benzyldimethylalkylammonium chloride by Aeromonas hydrophila sp. K. AB - AIMS: The aim of this work was to study the biodegradation of benzyldimethylalkylammonium chloride (BAC) by Aeromonas hydrophila sp. K, an organism isolated from polluted soil and capable of utilizing BAC as sole source of carbon and energy. METHODS AND RESULTS: High performance liquid chromatography and gas chromatography-mass spectrometry (GC-MS) analysis was used to study BAC degradation pathway. It was shown that during BAC biodegradation, formation of benzyldimethylamine, benzylmethylamine, benzylamine, benzaldehyde and benzoic acid occurred. Formation of benzyldimethylamine as the initial metabolite suggested that the cleavage of Calkyl-N bond occurred as the first step of BAC catabolism. Liberation of benzylmethylamine and benzylamine likely resulted from subsequent demethylation reactions, followed by deamination with formation of benzaldehyde. Benzaldehyde was rapidly converted into benzoic acid, which was further degraded. CONCLUSIONS: Aer. hydrophila sp. K is able to degrade BAC. A degradation pathway for BAC and related compounds is proposed. SIGNIFICANCE AND IMPACT OF STUDY: These findings are significant for understanding biodegradation pathways of benzyl-containing quaternary ammonium compounds. PMID- 12534819 TI - Development of an immunomagnetic separation-polymerase chain reaction (IMS-PCR) assay specific for Enterocytozoon bieneusi in water samples. AB - AIMS: Microsporidia have become widely recognized as important human pathogens. Among Microsporidia, Enterocytozoon bieneusi is responsible for severe gastrointestinal disease. To date, no current therapy has been proven effective. Their mode of transmission and environmental occurrence are poorly documented because of the lack of detection methods that are both species-specific and sensitive. In this study, we developed a sensitive and specific molecular method to detect E. bieneusi spores in water samples. METHODS AND RESULTS: The molecular assay combined immunomagnetic separation (IMS) and polymerase chain reaction (PCR) amplification to detect E. bieneusi spores. A comparison was made of IMS magnetic beads coated with two different monoclonal antibodies, one specific for the Encephalitozoon genus that cross-reacts with E. bieneusi and the other specific only for the E. bieneusi species itself. CONCLUSIONS: Immunotech beads coated with the antibody specific for E. bieneusi were found to be the most effective combination. SIGNIFICANCE AND IMPACT OF THE STUDY: The highly specific IMS-PCR assay developed in this study provides a rapid and sensitive means of screening water samples for the presence of E. bieneusi spores. PMID- 12534820 TI - The response of Leuconostoc mesenteroides to low external oxidoreduction potential generated by hydrogen gas. AB - AIMS: The physiological consequences of low external oxidoreduction potential in Leuconostoc mesenteroides were investigated. METHODS AND RESULTS: Leuconostoc mesenteroides was grown under two initial oxidoreduction potential conditions (Eh7: +200 mV and -400 mV) using nitrogen and hydrogen as reducing agents. Growth was affected by Eh7; the lag phase increased from 1 h at an initial Eh7 of +200 mV to 6 h at an initial Eh7 of -400 mV; the maximum specific growth rate at -400 mV was 68% of the one observed at +200 mV. The NADH/NAD+ ratio and (NADH + NAD+) pool were independent of the external Eh7. CONCLUSIONS: This study shows that changing the external oxidoreduction potential from +200 to -400 mV has a strong effect on the Leuc. mesenteroides physiology. The constancy of the maximum carbon and energetic fluxes (qglu, qATP) under the two Eh7 conditions accompanied by the decrease of YX/S and YATP suggested the existence of an uncoupling phenomenon, namely that some catabolized glucose and hence ATP was not associated with biomass production. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper demonstrates the usefulness of taking into account, the effect of the oxidoreduction potential on the growth of Leuc. mesenteroides in the fermentation process. PMID- 12534821 TI - Antibiotic resistance and genotyping of clinical group B Salmonella isolated in Accra, Ghana. AB - AIMS: The purpose of this study was to investigate the antibiotic resistance and clonal lineage of serogroup B Salmonella isolated from patients suspected of suffering from enteric fever in Accra, Ghana. METHODS AND RESULTS: Serogroup B Salmonella were isolated from blood (n=28), cerebral spinal fluid (CSF) (n=1), or urine (n=2), and identified based on standard biochemical testing and agglutinating antisera. Isolates were examined for their susceptibility to ampicillin, chloramphenicol, tetracycline and trimethoprim-sulfamethoxazole. Most of the isolates could be classified as multiple-drug resistant. Furthermore, the genetic location of resistance genes was shown to be on conjugative plasmids. Genetic fingerprinting by plasmid profiling, enterobacterial repetitive intergenic consensus (ERIC)-PCR, and repetitive element (REP)-PCR were performed to determine the diversity among the isolates. Plasmid profiling discriminated five unique groupings, while ERIC-PCR and REP-PCR resulted in two and three groupings, respectively. CONCLUSIONS: A high rate of antibiotic resistance was associated with the Salmonella isolates and the genes responsible for the resistance are located on conjugative plasmids. Also, there appears to be minimal diversity associated with the isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: As a result of the increasing antibiotic resistance among bacteria of all genera, surveys to monitor microbial populations are critical to determine the extent of the problem. The inability to treat many infectious diseases with current antibiotic regimens should prompt the medical community to be more prudent with its antibiotic use. PMID- 12534822 TI - Diversity of moderately halophilic bacteria producing extracellular hydrolytic enzymes. AB - AIMS: The aim of this study was to determine the diversity of moderately halophilic bacteria with hydrolase activities. METHODS AND RESULTS: Screening bacteria from different hypersaline environments in South Spain led to the isolation of a total of 122 moderately halophilic bacteria able to produce different hydrolases (amylases, DNases, lipases, proteases and pullulanases). These bacteria are able to grow optimally in media with 5-15% salts and in most cases up to 20-25% salts. In contrast to strains belonging to previously described species, that showed very little hydrolase activities, environmental isolates produced a great variety of hydrolases. These strains were identified as members of the genera: Salinivibrio (55 strains), Halomonas (25 strains), Chromohalobacter (two strains), Bacillus-Salibacillus (29 strains), Salinicoccus (two strains) and Marinococcus (one strain), as well as eight non-identified isolates. CONCLUSIONS: Moderately halophilic bacteria are a source of hydrolytic enzymes such as amylases, DNases, lipases, proteases and pullulanases. SIGNIFICANCE AND IMPACT OF THE STUDY: Although most culture collection strains are not able to produce hydrolases, it has been shown that environmental isolates can produce these potentially biotechnological important enzymes. PMID- 12534823 TI - Initial characterization of new bacteria degrading high-molecular weight polycyclic aromatic hydrocarbons isolated from a 2-year enrichment in a two liquid-phase culture system. AB - AIMS: To characterize some polycyclic aromatic hydrocarbons (PAH)-degrading microorganisms isolated from an enriched consortium degrading high molecular weight (HMW) PAHs in a two-liquid-phase (TLP) soil slurry bioreactor, and to determine the effect of low molecular weight (LMW) PAH on their growth and HMW PAH-degrading activity. METHODS AND RESULTS: Several microorganisms were isolated from a HMW-PAH (pyrene, chrysene, benzo[a]pyrene and perylene) degrading consortium enriched in TLP cultures using silicone oil as the organic phase. From 16S rRNA analysis, four isolates were identified as Mycobacterium gilvum B1 (99% identity),Bacillus pumilus B44 (99% identity), Microbacterium esteraromaticum B21 (98% identity), and to the genus Porphyrobacter B51 (96% identity). The two latter isolates have not previously been associated with PAH degradation. Isolate B51 grew strongly in the interfacial fraction in the presence of naphthalene vapours and phenanthrene compared with cultures without LMW PAHs. Benzo[a]pyrene was degraded in cultures containing a HMW PAH mixture but pyrene had no effect on its degradation. The growth of isolates B1 and B21 was improved in the aqueous phase than in the interfacial fraction for cultures with naphthalene vapours. Pyrene was required for benzo[a]pyrene degradation by isolate B1. For isolate B21, pyrene and chrysene were degraded only in cultures without naphthalene vapours. CONCLUSION: Consortium enriched in a TLP culture is composed of microorganisms with different abilities to grow at the interface or in the aqueous phase according to the culture conditions and the PAH that are present. Naphthalene vapours increased the growth of the microorganisms in TLP cultures but did not stimulate the HMW PAH degradation. SIGNIFICANCE AND IMPACT OF THE STUDY: New HMW PAH-degrading microorganisms and a better understanding of the mechanisms involved in HMW PAH degradation in TLP cultures. PMID- 12534824 TI - Raw potato starch and short-chain fructo-oligosaccharides affect the composition and metabolic activity of rat intestinal microbiota differently depending on the caecocolonic segment involved. AB - AIMS: In vitro studies have suggested that fructo-oligosaccharides (FOS) and resistant starch (two fermentable non-digestible carbohydrates) display different fermentation kinetics. This study investigated whether these substrates affect the metabolic activity and bacterial composition of the intestinal microflora differently depending on the caecocolonic segment involved. METHODS AND RESULTS: Eighteen rats were fed a low-fibre diet (Basal) or the same diet containing raw potato starch (RPS) (9%) or short-chain FOS (9%) for 14 days. Changes in wet content weights, bacterial populations and metabolites were investigated in the caecum, proximal and distal colon and faeces. Both substrates exerted a prebiotic effect compared with the Basal diet. However, FOS increased lactic acid-producing bacteria (LAPB) throughout the caecocolon and in faeces, whereas the effect of RPS was limited to the caecum and proximal colon. As compared with RPS, FOS doubled the pool of caecal fermentation products, while the situation was just the opposite distally. This difference was mainly because of the anatomical distribution of lactate, which accumulated in the caecum with FOS and in the distal colon with RPS. Faeces reflected these impacts only partly, showing the prebiotic effect of FOS and the metabolite increase induced by RPS. CONCLUSIONS: This study demonstrates that FOS and RPS exert complementary caecocolonic effects. SIGNIFICANCE AND IMPACT OF THE STUDY: The RPS and FOS combined ingestion could be beneficial by providing health-promoting effects throughout the caecocolon. PMID- 12534825 TI - Discrimination of Staphylococcus aureus biotypes by pulsed-field gel electrophoresis of DNA macro-restriction fragments. AB - AIMS: To examine whether pulsed-field gel electrophoresis (PFGE) of DNA macro restriction fragments could provide better discrimination among the different biotypes previously described within the species Staphylococcus aureus than the traditional biochemical approach. METHODS AND RESULTS: Seventy three Staph. aureus strains from various sources (human, animal or food origin) and belonging to eight biotypes, including the poultry-like biotype, tentatively designated as an 'abattoir' biotype, were genotyped by PFGE after SmaI digestion of DNA. The PFGE patterns were compared using the average linkage matching method (UPGMA) with the Dice coefficient. A total of 61 PFGE patterns were observed, showing between 31 and 100% similarity. In most cases, strains with the same biotype were grouped specifically into one, two or three separate sub-clusters. Strains from the 'abattoir' biotype were clustered in one separate sub-cluster. CONCLUSIONS: The PFGE typing is useful to distinguish the traditional biotypes of Staph. aureus and has a more discriminatory power than the biochemical typing. SIGNIFICANCE AND IMPACT OF THE STUDY: The PFGE typing confirms the 'abattoir' biotype as a separate group on a genetic level and is well suited to investigate modes of staphylococcal contamination of food. PMID- 12534826 TI - Liquid formulation of the biocontrol agent Candida sake by modifying water activity or adding protectants. AB - AIMS: To evaluate the effect of modification of water activity (aw) and the addition of protective substances in the preservation medium of liquid formulations of the biocontrol agent Candida sake stored at 4 and 20 degrees C. METHODS AND RESULTS: The aw of the preservation medium of C. sake was modified from 0.72 to 0.95 by adding glycerol or polyethylene glycol (PEG). Moreover, several protectant substances at different concentrations were evaluated. Modification of lower aw-levels (0.721-0.901) with glycerol did not maintain the viability of the yeast cells. Higher aw-levels (0.93-0.95) with either glycerol or PEG improved the viability but not at acceptable viability levels. C. sake cells maintained viabilities >60% when sugars, such as trehalose, and polyols, such as glycerol and PEG were used as protectants in liquid formulations. Moreover, liquid formulations of C. sake stored at 4 degrees C showed higher number of viable counts than at 20 degrees C. When different sugars were tested, all of them, except 10% fructose, resulted in a viability higher than 50% of the C. sake formulations. Biocontrol of liquid formulation treatments was similar to fresh cells in controlling Penicillium expansum on wounded apples. CONCLUSIONS: Sugars such as lactose and trehalose could be considered as good protectants in order to obtain liquid formulations of C. sake cells as they maintain the viability >70% for 4 months at 4 degrees C. SIGNIFICANCE AND IMPACT OF STUDY: This study shows that a suitable liquid formulation for commercial application can be produced with high viability and conservation of biocontrol efficacy. Moreover, if 10% lactose is the protectant used in the formulation, the economic costs would not be limiting for industrial production. PMID- 12534827 TI - The pathogenicity of strains of Salmonella paratyphi B and Salmonella java. AB - AIMS: To relate the diseases caused by strains of Salmonella paratyphi B and S. java to pathogenic mechanisms expressed by these bacteria for the purpose of organism discrimination. METHODS AND RESULTS: Epidemiological data relating to cases of disease caused by strains of S. paratyphi B and S. java, isolated over a 10-year period, were analysed with respect to patients' symptoms, particularly those involving enteric fever. Strains of S. paratyphi B and S. java were also examined for a range of known pathogenic mechanisms. Infection with S. paratyphi B involved pyrexia in 12.5% of patients compared with 2.2% of patients infected with S. java. These organisms could not be differentiated based on the pathogenic properties examined. CONCLUSIONS: Strains of S. paratyphi B appear not to be a major cause of enteric fever but primarily a cause of gastroenteritis, in common with S. java. Both organisms express similar pathogenic mechanisms, and strains of S. java are probably d-tartrate utilizing variants of S. paratyphi B. SIGNIFICANCE AND IMPACT OF THE STUDY: Strains of S. paratyphi B are very closely related organisms, primarily causing gastroenteritis. From this study it would appear that strains of S. paratyphi B are not a major cause of enteric fever. PMID- 12534828 TI - Aspects of development of digestive activity of intestine in young chickens, ducks and geese. AB - The experiment comprised 100 chickens, 100 ducks and 100 geese fed diets based on mixed grains of maize, barley and wheat. Nutrient concentrations in the mixtures were in accordance with the requirements of poultry species. The birds were kept in metabolic cages from the 1st up to the 42nd day of life. The measurements of body weight, activity of alpha-amylase and lipase in pancreatic tissue, enzymatic activity of intestinal wall cells, parameters of carbohydrate fermentations, ileal and total amino acids digestibility were conducted on the 1st, 3rd, 5th, 7th, 28th and 42nd day of life or on the 14th, 28th and 42nd day of life. The pancreas alpha-amylase activity in chickens during the whole period was quite constant; in ducks and geese it was very low, but from the 28th day a dynamic increase in the activity of the pancreatic enzymes was observed in these birds. The activity of lipase was low but from the 28th day of experiment an increase was noted and was much higher in waterfowl than in chickens. The highest caecal concentration of SCFA (Short chain fatty acids) (in total high) was noted in chickens; these values were slightly lower in geese and ducks. A high (70%) ileal amino acids digestibility was seen in very young chickens; during the period of 15-42 days this value amounted up to 73%. In geese, the obtained values were 63% and in ducks 43-61%, with a mean of 55%. Faecal digestibility of amino acids had a mean of up to 86% for all species but the digestibility of single amino acids such as cystine, glycine, histidine and tyrosine was diversified and relatively lower than the others. PMID- 12534829 TI - Reproduction of the raccoon dog (Nyctereutes procyonoides) after feeding or food deprivation in winter. AB - The wild raccoon dog (Nyctereutes procyonoides, Canidae, Carnivora) goes through autumn fattening followed by winter sleep. Farmed raccoon dogs also exhibit autumn fattening but not winter sleep, as a result of daily feeding and lack of nests. We studied the effects of food deprivation and winter sleep or active winter feeding on the physiology and reproduction of farm-born raccoon dogs. Eighty-six animals were put on a 2-month fast in November-December. The fast caused no deleterious effects on the health of the raccoon dogs. In the spring the food-deprived animals had slightly more cubs per mated female than the fed animals. There was a significant negative correlation between the number of cubs obtained and the mean body mass of the females at the beginning of the mating season. The highest mean number of cubs was obtained by the females that weighed 5-7 kg. The results indicate that the raccoon dog is finely adapted to a long period of food deprivation in the winter. Furthermore, winter sleep and food deprivation could be introduced to farm conditions by providing the raccoon dogs with nestboxes and withholding food for a period of 6-8 weeks in mid-winter. PMID- 12534830 TI - Influence of high levels of vitamin E on semen parameters of cocks. AB - This study was an attempt to find whether the reproductive performance of cocks would be influenced by oral administration of different amounts of dietary vitamin E over a long period of time. For that purpose 60 cocks were divided into five dietary groups of 12 animals each, and supplemented with 0 (control group), 100, 1,000, 10,000 or 20,000 IU alpha-tocopherol/kg diet, respectively, over a period of 12 months. The effect on semen parameters and biochemical parameters measured in pooled semen samples and the weight of the testes were determined. The weight of testes decreased with increasing amounts of supplemented vitamin E. Volume, pH, colour, consistency and motility were not influenced by the diets, but density of ejaculate (sperm/microl), total amount of spermatozoa and morphology of sperm were significantly lowered by increasing amounts of supplemented vitamin E. The alpha-tocopherol concentration in ejaculates increased significantly in relation to the diet whereas phospholipid content and thiobarbituric acid-reactive substances (TBARS) of the semen samples decreased significantly with increasing vitamin E supplementation. The reproductive performance of cocks was negatively influenced by high doses of vitamin E although decreased TBARS indicated rising oxidative defence. PMID- 12534831 TI - Factors affecting ascorbic acid biosynthesis in chickens: III. Effect of dietary fluoride on L-gulonolactone oxidase activity and tissue ascorbic acid (AsA) concentration. AB - The inconsistent beneficial responses to dietary ascorbic acid (AsA) may be due to dietary factors that alter biosynthesis or tissue turnover of AsA. It has been suggested on the basis of altered tissue AsA that dietary fluoride is a determinant of biosynthesis in chickens. Fluoride may enter the food chain of poultry via industrial contamination, feed ingredients and drinking water. The goal of this study was to ascertain whether dietary fluoride at 300 mg/kg influences l-gulonolactone oxidase (GLO) activity in commercial meat-type chickens. The experimental diet was fed from day-old to 3 weeks and responses measured. Growth and feed conversion were not affected by fluoride in the diet. Dietary fluoride neither inhibited nor enhanced GLO activity nor did it increase or decrease AsA concentration in plasma, liver, kidney, adrenal gland and muscle (pectoralis major). Tissue AsA concentration in ascending order was adrenal > liver > kidney > pectoralis major > plasma. The results are consistent with that reported for the rat and calculations based on the results eliminate fluorine contamination for the inconsistent responses of immature chickens to dietary AsA. PMID- 12534832 TI - Effects on health and blood plasma parameters of laying hens by pure nivalenol in the diet. AB - The effects of trichothecene nivalenol (NIV) on the health and production of laying hens were determined in a feeding study. White Leghorn hens, 55 weeks old, were fed diets containing 0, 1, 3 and 5 mg NIV/kg during 50 days. Feed intake was reduced by NIV, but there were no effects on body weight, egg production and egg quality. Traces of unaltered NIV were found in liver and bile. Alkaline phosphatase in the blood plasma increased in the hens fed 0, 1 and 3 mg NIV/kg diet but decreased in the birds fed 5 mg NIV/kg feed. Total protein and glucose were slightly reduced in hens fed 5 mg NIV/kg diet. Pathological examination revealed that 40-75% of hens fed NIV (3 and 5 mg/kg) supplemented diet showed gizzard lesions, haemorrhages in the duodenum and swollen cloaca and oviducts with immature eggs while certain birds in the 1 mg NIV group showed light and fragile livers. NIV and the metabolite de-epoxy-NIV in faeces were found to constitute up to 10% of ingested NIV. PMID- 12534833 TI - Energy and protein metabolism in pregnant sows fed two levels of dietary protein. AB - The present study was performed to quantify the energy and nutrient metabolism of pregnant sows fed high (HP) or low (LP) dietary protein [18.3 vs. 13.5% of dry matter (DM)]. A total of nine sows (four on HP and five on LP diet) were subjected to balance and respiration trials four times during their second pregnancy (approximately on days 30, 61, 80 and 104 of gestation). The digestibility of protein (83.0 vs. 79.9) (p < 0.01) and energy (84.9 vs. 83.7%) (p < 0.05) was higher for the HP diet. Daily intake of metabolizable energy (ME) and retained energy (RE) were similar for the two groups, with an average of 28.37 MJ ME and 3.94 MJ RE, respectively. Heat production (HE) measured according to the respiratory quotient (RQ) and carbon-nitrogen (CN) method was similar (464 vs. 454 kJ/kg 0.75/day, respectively). Sows fed HP retained more energy in protein (3.33 vs. 2.00 MJ/day) (p < 0.001) and tended to retain less energy in fat (1.59 vs. 2.50 MJ/day) than LP sows. Retained nitrogen (N) (22.3 vs. 13.4 g/day) (p < 0.001) and utilization of N (retained/digested) (45.2 vs. 38.1%) was higher for HP sows compared with LP sows. In late pregnancy, retained N, retained fat, HE and oxidation of carbohydrates increased, while oxidation of fat was reduced to zero. In conclusion, both diets provided adequate N for retention in maternal tissue and conception products. In spite of the lower utilization of N in LP sows, the N excretion was depressed by 5.6 g/day compared with HP sows, because of the lower N intake. PMID- 12534834 TI - Adaptation to the digestion of nutrients of a starch diet or a non-starch polysaccharide diet in group-housed pregnant sows. AB - A trial was conducted with twenty group-housed pregnant sows to study the adaptation in nutrient digestibility to a starch-rich diet or a diet with a high level of fermentable non-starch polysaccharides (NSP) during a time period of 6 weeks. The starch-rich diet was primarily composed of wheat, peas and tapioca, whereas soya bean hulls and sugar beet pulp, which both are highly fermentable NSP sources, were used to formulate the NSP-rich diet. The starch-rich diet and the NSP-rich diet were formulated to contain different levels of starch (298 vs. 78 g/kg) and fermentable NSP (100 vs. 300 g/kg), but a similar level of net energy (NE) (8.36 MJ/kg). The trial consisted of a 1-week adaptation period followed by a 5-week collection period. Weekly apparent faecal digestibilities of dry matter (DM), crude protein (CP), crude fat, ash and NSP were measured by using the acid-insoluble ash marker method. Apparent faecal digestibilities of DM and organic matter (OM) of both diets were similar. Faecal digestibility of CP and crude fat was lower (p < 0.001) whereas that of NSP was higher (p < 0.001) for sows that received the NSP-rich diet. Calculated NE values of both diets were similar. Sows fed the NSP-rich diet produced faeces that contained a lower (p < 0.001) DM content compared with sows that were fed with the starch-rich diet. The quantity of dry faeces was the same on both diets, therefore total faeces production (as-is basis) was higher (p < 0.01) for the sows fed the NSP-rich diet. During the 5-week collection period, no changes were observed in the digestibility of DM, OM and NSP in the NE value of the diets. Digestibilities of CP and fat, however, were lower in week 1 (p < 0.05) compared with weeks 2-5 for both diets. The DM content of the faeces and the quantity of dry faeces did not change from weeks 1-5. Diet by time interaction was not observed for any of the response variables indicating that sows adapt as quickly to a diet with a high level of fermentable NSP as to a starch-rich diet. The present trial shows that, with regard to digestibility of nutrients, pregnant sows completely adapt to a NSP-rich diet (i.e., NSP from sugar beet pulp) in 2 weeks and that the time period necessary to adapt to a starch-rich diet or a diet with a high level of fermentable NSP is similar. PMID- 12534835 TI - Characterization of 18:1 and 18:2 isomers produced during microbial biohydrogenation of unsaturated fatty acids from canola and soya bean oil in the rumen of lactating cows. AB - Ruminal production of biohydrogenation intermediates in response to unsaturated oils was assessed using 24 Jersey cows fed a control diet or the control diet supplemented at 35 g/kg dry matter (DM) with canola, soya bean, or a mixture of equal amounts of canola plus soya bean oil for 4-weeks. Total fatty acid content averaged 63 or 35 g/kg DM for oil-supplemented diets or control. Oleic acid accounted for 6, 29, 21 or 12 g/kg DM in the control, canola, mixture, or soya bean oil diet, respectively. Linoleic acid averaged 17, 19, 26, or 33 g/kg DM and linolenic acid 5, 5, 6 or 8 g/kg DM for control, canola, mixture, or soya bean oil. Concentrations of cis12-, trans11-, trans13+14, and trans15-18:1 were 0.81, 2.99, 2.24, and 0.73 mg/g rumen fluid, respectively, in response to soya bean oil and were 126, 90, 45, and 38% greater compared with other diets. Trans11cis15-, cis9trans11- and cis9 cis11-18:2 also were greater when soya bean oil (0.30, 0.34 and 0.01 mg/g, respectively) was fed compared with other treatments (0.12, 0.21 and 0.004 mg/g, respectively). Feeding canola oil resulted in greater concentrations of trans4-, trans5-, trans6+7+8-, trans9- and trans10-18:1 (0.20, 0.25, 0.87, 0.39 and 0.70 mg/g, respectively) compared with other diets (0.09, 0.15, 0.36, 0.20 and 0.46 mg/g, respectively). Trans10cis12-18:2 concentration did not differ as a result of diet and averaged 0.002 mg/g rumen contents. The pattern of 18:1 and 18:2 isomers formed during ruminal biohydrogenation depends greatly on dietary profile of unsaturated fatty acids. PMID- 12534838 TI - Delirium symptoms in post-acute care: prevalent, persistent, and associated with poor functional recovery. AB - OBJECTIVES: To determine the prevalence of delirium symptoms at the time of admission to post-acute facilities, the persistence of delirium symptoms in this setting, and the association of delirium symptoms with functional recovery. DESIGN: Prospective cohort study. SETTING: Eighty-five post-acute care facilities: 55 rehabilitation hospitals and 30 skilled nursing facilities in 29 states. PARTICIPANTS: Five hundred fifty-one consenting patients aged 65 and older newly admitted to participating facilities from acute care hospitals. MEASUREMENTS: Data were collected as part of a field study effort related to the Minimum Data Set (MDS). Basic demographic data, medical comorbidity, delirium symptoms, and functional status--activities of daily living (ADLs) and instrumental activities of daily living (IADLs)--were obtained from MDS assessments performed within 4 days of admission and again 1 week later by the patient's primary nurse. Six delirium symptoms (easily distracted, periods of altered perception, disorganized speech, periods of restlessness, periods of lethargy, and mental function varies over the course of a day) were assessed after appropriate training. RESULTS: Of the 551 patients (mean age +/- standard deviation 78 +/- 7, 64% women), 126 had delirium symptoms on post-acute admission, for an overall prevalence of 23%. In patients with delirium symptoms on the admission assessment, 1 week later, 14% had completely resolved, 22% had fewer delirium symptoms, 52% had the same number of symptoms, and 12% had more symptoms. Of those with no delirium symptoms on admission, 4% had new symptoms 1 week later. Patients who had the same number of or more delirium symptoms at the second assessment had significantly worse ADL and IADL recovery than those with fewer or resolved delirium symptoms or those with no delirium symptoms at either assessment. Persistent delirium symptoms remained significantly associated with worse ADL and IADL recovery after adjusting for age, comorbidity, dementia, and baseline functional status. CONCLUSIONS: The data from this study provide strong preliminary evidence that, in patients newly admitted to post-acute care facilities from acute care hospitals, delirium symptoms are prevalent, persistent, and associated with poor functional recovery. Educational efforts are warranted to help post-acute facility staff recognize and manage this common and morbid condition. PMID- 12534839 TI - Cost effect of managing methicillin-resistant Staphylococcus aureus in a long term care facility. AB - OBJECTIVES: The purpose of this study was to measure the total consumption of resources involved in the care of a long-term care facility (LTCF) resident infected with methicillin-resistant Staphylococcus aureus (MRSA). DESIGN: A retrospective cohort study. SETTING: A 375-bed LTCF that provides two levels of care. PARTICIPANTS: Ninety LTCF residents infected with Staphylococcus aureus (mean age +/- standard deviation for methicillin-sensitive Staphylococcus aureus (MSSA) patients = 85 +/- 8.8, for MRSA patients = 82 +/- 9.5, P =.127; 49 MSSA and 41 MRSA patients). Inclusion criteria consisted of identification of a positive S. aureus culture in addition to symptoms/signs consistent with infection. Patients colonized with S. aureus were excluded. MEASUREMENTS: A standardized data collection tool was used to conduct chart and database review throughout the defined infection period. The type of information collected included demographic, infection characterization, antibiotic regimen, resource assessment, and cost data. The cost data were further categorized into total pharmaceutical, infection management, physician care, nursing care, and total infection cost. RESULTS: One hundred eleven cases were identified, with 90 cases eligible for evaluation. No difference in population demographics was noted between groups. A significantly higher number of patients in the MRSA group had an indwelling device (P <.001), pressure ulcer(s) (P =.028), or diabetes mellitus (P =.007). There was a significantly higher number of patients with congestive heart failure in the MSSA group (P =.047), but no difference existed in the primary infection site (P =.297) or the incidence of patients with more than two comorbidities (P =.509). The infection characterization variables included were also similar between groups. The most prevalent infection site was the urinary tract (48%) followed by skin/skin structure (38%). Because the majority of patients (82%) developed infection at least 30 days after their LTCF admission, the infections may be considered to have been largely LTCF acquired. The median infection management cost of an MRSA infection was six times greater than that of a MSSA infection (P <.001), whereas the median associated nursing care cost was two times greater (P =.001). The median overall infection cost associated with MRSA was 1.95 times greater than that of MSSA (median (range): MSSA 1,332 US dollars (268-7,265 US dollars) vs MRSA 2,607 US dollars (849-8,895 US dollars), P <.001). Nursing care cost constituted the major portion of the overall infection cost in both groups (MSSA 51%, MRSA 48%). Evaluation of antimicrobial management revealed that infected residents were treated with a wide array of combination therapies (65% of patients received combination therapy). CONCLUSIONS: The management of a resident infected with MRSA was much more costly to the LTCF than that of an MSSA-infected patient. The general care of the patient and not the specific antibiotic regimen influenced the large difference in cost between groups. The approach to the antibiotic management of these patients was variable. A more streamlined approach to infection management that facilitates a faster cure rate may dramatically lower resource consumption and improve patient outcomes. PMID- 12534840 TI - Pneumonia versus aspiration pneumonitis in nursing home residents: diagnosis and management. AB - OBJECTIVES: To determine the frequency of aspiration pneumonitis in nursing home residents with an initial diagnosis of pneumonia and to compare the clinical characteristics, management, and outcome of aspiration pneumonitis with those of pneumonia. DESIGN: Retrospective chart review. SETTING: Hospital geriatric unit for nursing home residents. PARTICIPANTS: Nursing home residents admitted to the inpatient geriatric unit with suspected pneumonia between May 1999 and April 2001 (n = 195 episodes). MEASUREMENTS: Aspiration events were defined as definite (witnessed or unwitnessed) or suspected. Aspiration pneumonitis was defined as symptoms/signs of lower respiratory tract infection plus a history of an aspiration event plus a lower lobe infiltrate on chest radiograph. Pneumonia was defined as symptoms/signs of lower respiratory tract infection plus an infiltrate on chest radiograph plus no history of an aspiration event. RESULTS: The 195 episodes were stratified into three clinical groups: aspiration pneumonitis (n = 86; aspiration history/infiltrate), pneumonia (n = 43; no aspiration history/infiltrate), and an aspiration event (n = 66; aspiration history/no infiltrate). In general, symptoms, signs, and laboratory tests were not useful in distinguishing between the three groups. Survivors with aspiration pneumonitis (13/75 (17%)) or with an aspiration event (20/60 (33%)) were significantly more likely not to be treated with an antibiotic or to be treated for 1 day or less than those with pneumonia (0/41; P <.001). Excluding those not treated, significantly more patients with pneumonia (33/40 (83%)) were discharged on antibiotic treatment than those with aspiration pneumonitis (35/70 (50%)) or an aspiration event (21/51 (41%); P <.001). There was no significant difference in hospital mortality between the three clinical groups. CONCLUSIONS: The findings of this study have implications for the diagnosis and management of suspected pneumonia in nursing home residents but require prospective validation. PMID- 12534841 TI - Identifying older people at risk of abuse during routine screening practices. AB - OBJECTIVES: To examine the association between various characteristics of community-based older people and a constructed measure of potential elder abuse. DESIGN: Cross-sectional design. SETTING: Public community-based long-term care programs in Michigan. PARTICIPANTS: Individuals aged 60 and older seeking home and community-based services in Michigan between November 1996 and October 1997 (N = 701). MEASUREMENTS: Data were collected using the Minimum Data Set for Home Care (MDS-HC) assessment. The dependent variable is a constructed measure of potential elder abuse reflecting physical and emotional abuse and neglect. Independent variables include demographic characteristics; diagnoses; behavioral measures; and cognitive, physical, and social functioning. RESULTS: Several measures of social support and social function were strongly associated with the signs of a potentially abusive environment: brittle support (odds ratio (OR) = 3.5, 90% confidence interval (CI) = 1.5-8.1), older person feels lonely (OR = 2.4, 90% CI = 1.3-4.5), and older person expresses conflict with family/friends (OR = 2.3, 90% CI = 1.2-4.3). Home care participants' alcohol abuse, psychiatric illness, lack of ease interacting with others, and short-term memory problems were also significantly associated with the signs of potential elder abuse. CONCLUSIONS: The results of this study suggest that the signs of potential elder abuse are associated with a diminishing social network and poor social functioning, although some characteristics of the older person's health are contributing factors. Improved understanding of the link between those characteristics and potential abuse will help healthcare providers, case managers, and others identify older people at high risk of abuse. PMID- 12534842 TI - Amnestic behavior in dementia: symptoms to assist in early detection and diagnosis. AB - OBJECTIVES: This study evaluated two amnestic behavior changes (repetitive questioning and repetitive actions) to determine their utility in screening for early dementia. DESIGN: Patient data were collected through a retrospective chart review. Comparison data from nondemented older people were collected prospectively from acquaintances of clinic patients. SETTING: The setting was a hospital-based outpatient memory disorder clinic. PARTICIPANTS: Participants were older individuals with no cognitive impairment (n = 25), undetermined dementia (n = 50), and definite dementia (n = 25). The undetermined cases were followed for at least 1 year to assess for conversion to dementia. MEASUREMENTS: Amnestic behaviors were assessed using informant-report for all participants. The behaviors were examined for their ability to distinguish between definite dementia cases and noncases. They were further evaluated for the ability to differentiate undetermined cases that eventually converted to definite dementia from cases that did not convert. RESULTS: Results indicated that repetitive behaviors were common in early and more-severe dementia cases. Repetitive behaviors were relatively uncommon in cognitively intact older participants. In analyses of the full study sample, engagement in repetitive behaviors had high sensitivity (0.97) in identifying dementia cases. CONCLUSIONS: Assessing repetitive behaviors in patients may be a useful means for family members and primary care physicians to screen for early cognitive impairment. The assessment can help to identify individuals that should be evaluated further for dementia. PMID- 12534843 TI - Bone loss predicts subsequent cognitive decline in older women: the study of osteoporotic fractures. AB - OBJECTIVES: To determine whether the rate of bone loss predicts subsequent cognitive decline independently of baseline bone mass and whether apolipoprotein E (ApoE) genotype explains the association. DESIGN: A prospective cohort study. SETTING: Clinical centers in Baltimore, Maryland; Minneapolis, Minnesota; Pittsburgh, Pennsylvania; and Portland, Oregon. PARTICIPANTS: Four thousand four hundred sixty-two women aged 70 and older (mean = 75.8) participating in the Study of Osteoporotic Fractures. MEASUREMENTS: Total hipbone mineral density (BMD) was measured 2 and 6 years after enrollment (mean follow-up = 3.5 years), and expressed as annualized percentage rate of bone change. A modified Mini Mental State Examination (mMMSE) was administered at 6 and 10 years (mean follow up = 4.5 years) and defined cognitive decline as a decline of three or more points on repeat mMMSE score. ApoE genotype information was available on 883 women. RESULTS: Cognitive decline occurred in 12% of the women with the least bone loss (by quartile), 14% in the second, 16% in the third, and 20% in those with the greatest bone loss. After adjustment for age, education, stroke, functional status, estrogen use, body mass index, and smoking, the results were similar. Those who lost the most BMD were almost 40% more likely than women in the lowest quartile to develop cognitive decline in the multivariate model (odds ratio (OR) = 1.4, 95% confidence interval (CI) = 1.1-1.8). A similar association between hipbone loss and cognitive decline was observed in the multivariate model further adjusting for ApoE e4 (OR = 1.5, 95% CI = 0.8-2.7). CONCLUSIONS: Women with more rapid hipbone loss were more likely to develop cognitive decline than those who had lower rate of loss (or who gained bone mass). Differences in functional status, estrogen use, and ApoE did not explain this association. Further investigation is needed to determine the mechanisms that link osteoporosis and cognitive decline. PMID- 12534844 TI - Is there a relationship between excessive drinking and functional impairment in older persons? AB - OBJECTIVES: To investigate the relationship between (1) two thresholds of excessive alcohol drinking, (2) binge drinking, and (3) impairments in functional status in older drinkers. DESIGN: Cross-sectional study. SETTING: Ten internal medicine practices affiliated with an academic medical center. PARTICIPANTS: One hundred sixty-one persons aged 60 and older who reported drinking one or more drinks in the previous 3 months. MEASUREMENTS: Two commonly used thresholds of excessive drinking: (1) eight to 14 drinks per week for women and men (lower threshold) and (2) more than 14 drinks per week for women and men (higher threshold); a measure of binge drinking (> or =3 drinks per occasion for women or > or =4 drinks per occasion for men); and self-reported instrumental activities of daily living (IADLs) and advanced activities of daily living (AADLs). RESULTS: Compared with older persons consuming seven or fewer drinks per week, those exceeding the higher threshold of excessive drinking were more likely to have impairments in IADLs (adjusted odds ratio (AOR) = 8.4) and, to a lesser extent, AADLs (AOR = 3.7); those exceeding the lower threshold were more likely to have impairments in IADLs (AOR 5 6.0) but not in AADLs (AOR = 1.7). Binge drinkers were also more likely to have impairments in IADLs (AOR = 3.0) but not in AADLs (AOR = 1.5). CONCLUSIONS: In this group of older men and women, drinking more than seven drinks per week was associated with impairments in IADLs and, to a lesser extent, AADLs. Drinking more than three drinks per occasion was associated with impairments in IADLs. PMID- 12534845 TI - Risk of hip fracture in disabled community-living older adults. AB - OBJECTIVES: To determine the rate of hip fracture and risk factors associated with hip fractures in disabled older persons who enroll in the Program of All Inclusive Care for the Elderly (PACE), a program providing comprehensive care to community-living nursing-home-eligible persons. DESIGN: Prospective cohort study between January 1990 and December 1997. SETTING: The twelve PACE demonstration sites: San Francisco, California; Columbia, South Carolina; Detroit, Michigan; Denver, Colorado; East Boston, Massachusetts; El Paso, Texas; Milwaukee, Wisconsin; Oakland, California; Portland, Oregon; Rochester, New York; Sacramento, California; and the Bronx, New York. PARTICIPANTS: Five thousand one hundred eighty-seven individuals in PACE; mean age 79, 71% female, 49% white, 47% with dementia. MEASUREMENTS: Functional status, cognitive status, demographics, and comorbid conditions were recorded on all the participants, who were tracked for occurrence of a hip fracture. The goals were to determine the rate of hip fracture and identify risk factors. RESULTS: Two hundred thirty-eight hip fractures (4.6%) occurred during follow-up. The rate of hip fracture was 2.2% per person-year. Four independent predictors of hip fracture were identified using Cox proportional hazard analysis: age of 75 and older (adjusted hazard ratio (HR) = 2.0, 95% confidence interval (CI) = 1.4-2.8); white ethnicity (HR = 2.1, 95% CI = 1.6-2.8); ability to transfer independently to and from bed, chair, and toilet (HR = 3.0, 95% CI = 1.2-7.2); and five or more Short Portable Mental Status Questionnaire errors (HR = 1.6, 95% CI = 1.3-2.1). The incidence of hip fracture ranged from 0.5% per person-year in persons with zero to one independent risk factors to 4.7% per person-year in those with all four independent risk factors. CONCLUSIONS: The rate of hip fracture in this cohort of disabled community-living older adults was similar to that reported in nursing home cohorts. Older age, white race, ability to transfer independently, and cognitive impairment were independent predictors of hip fracture. Persons with these risk factors should be targeted for preventive interventions, which should include strategies for making transferring safer. PMID- 12534846 TI - Adherence to pressure ulcer prevention guidelines: implications for nursing home quality. AB - OBJECTIVES: This study aims to assess overall nursing home (NH) implementation of pressure ulcer (PU) prevention guidelines and variation in implementation rates among a geographically diverse sample of NHs. DESIGN: Review of NH medical records. SETTING: A geographically diverse sample of 35 Veterans Health Administration NHs. PARTICIPANTS: A nested random sample of 834 residents free of PU on admission. MEASUREMENTS: Adherence to explicit quality review criteria based on the Agency for Healthcare Research and Quality Practice Guidelines for PU prevention was measured. Medical record review was used to determine overall and facility-specific adherence rates for 15 PU guideline recommendations and for a subset of six key recommendations judged as most critical. RESULTS: Six thousand two hundred eighty-three instances were identified in which one of the 15 guideline recommendations was applicable to a study patient based on a specific indication or resident characteristic in the medical record. NH clinicians adhered to the appropriate recommendation in 41% of these instances. For the six key recommendations, clinicians adhered in 50% of instances. NHs varied significantly in adherence to indicated guideline recommendations, ranging from 29% to 51% overall adherence across all 15 recommendations (P <.001) and from 24% to 75% across the six key recommendations (P <.001). Adherence rates for specific indications also varied, ranging from 94% (skin inspection) to 1% (education of residents or families). Standardized assessment of PU risk was identified as one of the most important and measurable recommendations. Clinicians performed this assessment in only 61% of patients for whom it was indicated. CONCLUSIONS: NHs' overall adherence to PU prevention guidelines is relatively low and is characterized by large variations between homes in adherence to many recommendations. The low level of adherence and high level of variation to many best-care practices for PU prevention indicate a continued need for quality improvement, particularly for some guidelines. PMID- 12534847 TI - Value of exercise capacity and heart rate recovery in older people. AB - OBJECTIVES: To evaluate the prognostic value in older adults of two predictors of mortality: impaired functional capacity and an attenuated heart rate recovery. SETTING: Academic medical center. DESIGN: Prospective study with mean 3.7 years follow-up. PARTICIPANTS: Seven thousand three hundred fifty-four adults aged 65 and older consecutively referred for exercise testing between 1990 and 1999. Patients with heart failure, valvular disease, atrial fibrillation, and pacemakers were excluded. MEASUREMENTS: The primary endpoint was all-cause mortality. Impaired functional capacity was defined as the peak exercise workload in the lowest quintile of metabolic equivalents achieved according to prespecified strata of age and sex. Heart rate recovery was defined as the fall in heart rate during the first minute after exercise and was abnormal if 12 or fewer beats per minute, except for patients undergoing stress echocardiography, in which case 18 or fewer beats per minute was abnormal. RESULTS: There were 842 deaths. Patients with impaired functional capacity were at increased risk for death (23% vs 9%, hazard ratio (HR) = 2.7, 95% confidence interval (CI) = 2.2 3.1, P <.0001) as were patients with an abnormal heart rate recovery (17% vs 9%, HR = 2.0, 95% CI = 1.8-2.3, P <.0001). After adjusting for age, sex, coronary history, and other confounders, impaired functional capacity (adjusted HR = 2.1, 95% CI = 1.8-2.4) and an abnormal heart rate recovery (adjusted HR = 1.5, 95% CI = 1.3-1.7) independently predicted death. No interactions between these two variables with age were noted. CONCLUSIONS: In older patients, impaired functional capacity and heart rate recovery were independent predictors of death. PMID- 12534848 TI - Family members' preferences for nutrition interventions to improve nursing home residents' oral food and fluid intake. AB - OBJECTIVES: To measure family members' preferences for nutrition interventions to improve the oral food and fluid intake of their relatives in a nursing home. DESIGN: Cross-sectional descriptive. SETTING: Three skilled nursing facilities in Southern California. PARTICIPANTS: One hundred five residents from the three skilled nursing facilities and their respective family members. MEASUREMENTS: A mailed questionnaire to family members that consisted of 15 forced-choice comparisons between six nutrition interventions. An assessment of oral food and fluid intake during mealtime for 3 days (nine meals) for each participant using direct observations and estimations of percentage consumed (0% to 100%) by trained research staff. RESULTS: In order of most to least desirable, the family members preferred the following interventions to improve their relative's oral food and fluid intake: (1) improve quality of food; (2) improve quality and quantity of feeding assistance; (3) provide multiple small meals and snacks throughout the day; (4) place resident in preferred dining location; (5) provide an oral liquid nutritional supplement between meals; and (6) provide a medication to stimulate appetite. The average +/- standard deviation total percentage intake for residents whose family members reported that they thought their relative had a problem with their intake was 50%+/- 16%. CONCLUSIONS: Family members prefer that other nutrition interventions be attempted before the use of oral supplements or pharmacological approaches. Family members perceive a need for interventions when residents consume, on average, only half of the food and fluid items provided during mealtime. PMID- 12534849 TI - Nursing home characteristics associated with tube feeding in advanced cognitive impairment. AB - OBJECTIVES: To identify nursing homes factors associated with the use of tube feeding in advanced cognitive impairment. DESIGN: Descriptive study. SETTING: The On-line Survey Certification of Automated Records (OSCAR) was used to obtain facility characteristics from 1,057 licensed nursing homes in six states from 1995 to 1996. PARTICIPANTS: Residents aged 65 and older with advanced cognitive impairment who had a feeding tube placed over a 1-year period were identified using the Minimum Data Set. MEASUREMENTS: Nursing home characteristics independently associated with feeding tube placement were determined. RESULTS: Having a full-time speech therapist on staff, more licensed nurses and fewer nursing assistants were independently associated with greater use of tube feeding in severely cognitively impaired residents. Other features associated with tube feeding included larger facility size, higher proportion of Medicaid beds, absence of an Alzheimer's disease unit, pressure ulcers in 10% or more of residents, and a higher proportion of residents lacking advance directives and with total functional dependency. CONCLUSIONS: Assessment by a speech therapist, staffing ratios, advance directives, fiscal considerations, and specialized dementia units are potentially modifiable factors in nursing homes that may influence the practice of tube feeding in advanced cognitive impairment. PMID- 12534850 TI - Does inflammation or undernutrition explain the low cholesterol-mortality association in high-functioning older persons? MacArthur studies of successful aging. AB - OBJECTIVES: To explore the effect of inflammation and undernutrition on the association between hypocholesterolemia and higher overall mortality in high functioning older persons. DESIGN: Prospective cohort study. SETTING: Three U.S. communities. PARTICIPANTS: A cohort of 870 participants from the MacArthur Studies of Successful Aging. MEASUREMENTS: Baseline information was obtained for serum levels of cholesterol, C-reactive protein, interleukin-6, and albumin; body mass index; prevalent medical conditions; health behaviors; and medications. Crude and multivariate logistic regression analyses were used to examine the association between serum total cholesterol levels and 7-year all-cause mortality, while adjusting for potential confounders. RESULTS: In univariate analysis, the risk ratio of low serum total cholesterol level (<169 mg/dL) for 7 year total mortality was 1.90 (95% confidence interval (CI) = 1.18-3.07). The multiple adjusted risk ratios were 1.82 (95% CI = 1.10-3.00) after controlling for markers of inflammation and nutrition and 1.39 (95% CI = 0.80-2.40) after adjustment for additional cardiovascular risk factors. Sex was an important confounding variable that contributed to the observed inverse association between low serum cholesterol and overall mortality in univariate analysis. CONCLUSIONS: Hypocholesterolemia is not an independent risk factor for increased overall mortality in high-functioning community-dwelling older men and women. The association between low total cholesterol and high mortality observed in crude analysis is mainly confounded by common cardiovascular risk factors, rather than underlying inflammation or undernutrition. PMID- 12534851 TI - Malnutrition in institutionalized seniors: the iatrogenic component. AB - OBJECTIVES: To provide practitioners with benchmarks for micronutrient supplementation policies for older residents of long-term care (LTC) facilities, based upon residents' usual energy intake. DESIGN: Retrospective secondary analysis of nutrient intake and delivery data, obtained from 21 consecutive collection days. SETTING: A fully accredited geriatric teaching facility, affiliated with the University of Toronto's Medical School, that includes a chronic care hospital and a home for the aged, housing more than 800 senior residents. PARTICIPANTS: Twenty-three cognitively impaired residents who retained the ability to feed themselves. MEASUREMENTS: Average daily micronutrient (vitamin and mineral) content of two common diet types--unrestricted diet as tolerated and lactose-free--were evaluated based upon all foods served during a 28-day cycle of menus, providing an estimate of total possible micronutrient intake. Energy intake needed to meet recommended intakes for each of the micronutrients was then determined and compared with actual dietary intakes. RESULTS: Even if entirely consumed, neither diet (providing approximately 2,000 kcal/d) supplied sufficient quantities of vitamins (vitamin E, pantothenic acid) and minerals (calcium, zinc, copper and manganese) to enable residents to meet recommended intakes, making these deficiencies iatrogenic in nature. As projected intake levels fell to reflect ranges more consistently observed in LTC residents (1,000-1,500 kcal/d), the number and severity of risk nutrients increased. Similar predictions, based upon actual intake measurements, were made and confirmed. CONCLUSIONS: The traditional approach of developing LTC menus using resources such as Canada's Food Guide or the Food Guide Pyramid of the United States results in iatrogenic malnutrition. New guidelines are needed that are targeted toward the special needs of older adults who have low food intakes. In the interim, all older LTC residents require full-spectrum vitamin and mineral supplements. PMID- 12534852 TI - Development of a scale to predict decline in patients with mild Alzheimer's disease. AB - OBJECTIVES: To develop a scale that can assist in predicting likelihood of decline from mild dementia over 1 year in patients with Alzheimer's disease (AD). DESIGN: Retrospective cohort study. SETTING: University Memory and Aging Center. PARTICIPANTS: Patients with probable or possible AD and Clinical Dementia Rating (CDR) of 1 at baseline, divided into development and validation cohorts (n = 118 each). MEASUREMENTS: The CDR and neurological and neuropsychological assessments were given at baseline and 1 year later. RESULTS: In the development cohort, high education, low Mini-Mental State Examination score, poor insight, psychotic symptoms, and greater activity of daily living impairment predicted decline in CDR from 1 to 2 or 3. Receiver operating characteristics (ROC) curve analysis identified cutoff scores that maximized sensitivity and specificity for each significant predictor of decline. Based on the cutoff, raw scores were recoded to reflect risk for decline, weighted, and summed to create a final scale score. ROC curve analysis established a cutoff to indicate risk for decline on the final scale score. Sensitivity, specificity, and area under the ROC were 0.76, 0.74, and 0.83 in the development cohort and 0.77, 0.69, and 0.80 in the validation cohort, respectively. Positive and negative predictive values were 0.71 and 0.78 in the development cohort and 0.68 and 0.78 in the validation cohort, respectively. CONCLUSIONS: Decline from mild to moderate or severe impairment represents significant clinical change, with implications for patient and caregiver quality of life and treatment options. The clinical scale developed uses data to enhance prediction about change from mild to moderate or severe stages of AD. PMID- 12534853 TI - The MDS-CHESS scale: a new measure to predict mortality in institutionalized older people. AB - OBJECTIVES: To develop a scale predicting mortality and other adverse outcomes associated with frailty. DESIGN: Observational study based on Minimum Data Set (MDS) 2.0 and mortality data. SETTING: Ontario chronic hospitals. PARTICIPANTS: All chronic hospital patients (N = 28,495) assessed with the MDS 2.0 after mandatory implementation in July 1996 followed until May 1999. MEASUREMENTS: MDS 2.0 assessments done as part of normal practice mainly by registered nurses or multidisciplinary teams in a chronic hospital. Mortality data are available from the accompanying discharge tracking form. RESULTS: The MDS-Changes in Health, End stage disease and Symptoms and Signs (CHESS) score is a composite measure addressing changes in health, end-stage disease, and symptoms and signs of medical problems. It is a strong predictor of mortality (P <.0001) independent of the effects of age, sex, activities of daily living impairment, cognition, and do not-resuscitate orders. It is also strongly associated with physician activity, complex medical procedures, and pain (P <.001 for each dependent variable). CONCLUSIONS: The CHESS score provides a useful new MDS-based test to predict mortality and to measure instability in health as a clinical outcome. PMID- 12534854 TI - Testosterone supplementation therapy for older men: potential benefits and risks. AB - Serum testosterone levels decline gradually and progressively with aging in men. Many manifestations associated with aging in men, including muscle atrophy and weakness, osteoporosis, reduced sexual functioning, and increased fat mass, are similar to changes associated with testosterone deficiency in young men. These similarities suggest that testosterone supplementation may prevent or reverse the effects of aging. A MEDLINE search was performed to identify studies of testosterone supplementation therapy in older men. A structured, qualitative review was performed of placebo-controlled trials that included men aged 60 and older and evaluated one or more physical, cognitive, affective, functional, or quality-of-life outcomes. Studies focusing on patients with severe systemic diseases and hormone deficiencies related to specific diseases were excluded. In healthy older men with low-normal to mildly decreased testosterone levels, testosterone supplementation increased lean body mass and decreased fat mass. Upper and lower body strength, functional performance, sexual functioning, and mood were improved or unchanged with testosterone replacement. Variable effects on cognitive function were reported, with improvements in some cognitive domains (e.g., spatial, working, and verbal memory). Testosterone supplementation improved exercise-induced coronary ischemia in men with coronary heart disease, whereas angina pectoris was improved or unchanged. In a few studies, men with low testosterone levels were more likely to experience improvements in lumbar bone mineral density, self-perceived functional status, libido, erectile function, and exercise-induced coronary ischemia with testosterone replacement than men with less marked testosterone deficiency. No major unfavorable effects on lipids were reported, but hematocrit and prostate specific antigen levels often increased. Based on these results, testosterone supplementation cannot be recommended at this time for older men with normal or low-normal testosterone levels and no clinical manifestations of hypogonadism. However, testosterone replacement may be warranted in older men with markedly decreased testosterone levels, regardless of symptoms, and in men with mildly decreased testosterone levels and symptoms or signs suggesting hypogonadism. The long-term safety and efficacy of testosterone supplementation remain uncertain. Establishment of evidence-based indications will depend on further demonstrations of favorable clinical outcomes and symptomatic, functional, and quality-of-life benefits in carefully performed, long-term, randomized, placebo-controlled clinical trials. PMID- 12534855 TI - Telomeres, telomerase, and telomerase inhibition: clinical implications for cancer. AB - Telomeres are located at the ends of eukaryotic chromosomes. The enzyme telomerase synthesized them, and they are responsible for maintaining the lengths of chromosomes. Absence of telomerase is associated with telomere shortening and aging of somatic cells, but high telomerase activity is observed in over 90% of human cancer cells. Although the disappearance of telomerase with aging is considered a natural defense against development of cancer, it is not known what triggers the reappearance of telomerase in cancer cells. Telomerase activity is directly correlated with the expression of its active catalytic component, the human telomerase reverse transcriptase (hTERT), which is controlled primarily at the level of transcription. An earlier paper discussed the relationship of telomerase with aging. In this article, the contemporary literature is reviewed to explore the associations between telomerase, telomerase inhibition, and cancer. Because most cancers occur in old age, with the aging of the population, the number of people suffering from cancer is expected to increase in the coming decades. It is not known what roles telomerase and hTERT play in the complex relationship between aging and cancer. Data from experimental studies suggest that telomerase assay could potentially play a role in the diagnosis and prognosis of cancers. There is also evidence that telomerase inhibitors might be used as anticancer agents. As the knowledge of the relationships between telomerase and cancer and between telomerase and aging advances, it is hoped that more about the interacting relationships between telomerase, aging, and cancer will be learned. PMID- 12534856 TI - American College of Cardiology/American Heart Association Chronic Heart Failure Evaluation and Management guidelines: relevance to the geriatric practice. AB - BACKGROUND: Heart failure (HF) is the only cardiovascular disease with increasing incidence and prevalence. Most HF patients are older adults. With the aging of the population and effective treatment of hypertension and coronary artery disease, the two major underlying causes of HF, the number of older Americans with HF is expected to rise significantly in the coming decades. HF is the number one hospital discharge diagnosis for older adults. It is one of the causes of frequent hospital readmissions, reflecting acute decompensation and compromised quality of life for patients and increased cost and resource use for the healthcare system. It is also associated with approximately 300,000 deaths annually, most in older adults. Advances in the management of HF in the past several decades have significantly decreased the mortality and morbidity associated with this condition. Randomized controlled trials have demonstrated the beneficial effects of angiotensin-converting enzyme (ACE) inhibitors, angiotensin receptor blockers, beta-blockers, and spironolactone on survival and quality of life of HF patients, but there is evidence of underuse of evidence based care for HF. Several national guidelines have been published since 1994 that recommended evidence-based evaluation and management of HF. In 1995, the American College of Cardiologists (ACC) and the American Heart Association (AHA) published their first HF guidelines that recommended left ventricular (LV) function evaluation for all patients presenting with HF and use of ACE inhibitors for all patients with LV systolic dysfunction (LVSD) unless contraindicated. The guidelines recommended the use of hydralazine and isosorbide dinitrate in patients who could not use ACE inhibitors. In addition, digoxin was recommended in patients with HF due to LVSD but not adequately responsive to ACE inhibitors and diuretics and in those with atrial fibrillation and rapid ventricular rates. Diuretic use was recommended for symptomatic patients with evidence of fluid overload. Use of anticoagulation was restricted to patients with atrial fibrillation or to those with a history of systemic or pulmonary embolism. Beta blockers were reserved for HF patients after acute myocardial infarctions. Recent advances in the management of HF called for a revision of the guidelines. PURPOSE: The purpose of revising the 1995 ACC/AHA guidelines was to incorporate recent advances in pharmacological and nonpharmacological approaches to HF treatment and to assist physicians in clinical decision-making in the management of HF. METHODS: The ACC/AHA invited representatives from the American College of Chest Physicians, the Heart Failure Society of America, the International Society for Heart and Lung Transplantation, the American Academy of Family Physicians, and the American College of Physicians-American Society of Internal Medicine to participate in the preparation of the guidelines. The writing committee searched pertinent medical literature in English using computerized databases such as MEDLINE and EMBASE and manually searching the bibliographies of the selected articles. The writing committee classified HF into four stages, including patients who are at high risk for developing HF. According to this new classification, Stage A and B represent asymptomatic patients, the former at high risk for developing HF but no structural disorder, the latter with structural disorders of the heart. Stage C encompasses patients with underlying structural heart disorders who are symptomatic, either currently or in the past. Stage D includes end-stage HF patients. This new classification is intended to complement the New York Heart Association (NYHA) functional classification of HF into Class I (asymptomatic), Class II (symptomatic on less than ordinary exertion), Class III (symptomatic on ordinary exertion), and Class IV (symptomatic at rest). RESULTS: The recommendations are categorized into those for evaluation and management. PMID- 12534858 TI - Financial incentives for placing feeding tubes in nursing home residents with advanced dementia. PMID- 12534859 TI - Donepezil for nursing home patients with dementia: a reinterpretation of the evidence. PMID- 12534861 TI - Getting donepezil into the nursing home. PMID- 12534862 TI - Interdisciplinary care in orthogeriatrics: a good cost-benefit model of care. PMID- 12534864 TI - Capillary blood glucose testing in older people: reliable and accurate. PMID- 12534865 TI - International aging--India. PMID- 12534866 TI - Case report: a swollen and red ear. PMID- 12534867 TI - The Vulnerable Elders Survey: a tool for identifying vulnerable older people in the community. PMID- 12534868 TI - Apparent transmission of Citrobacter koseri in catheterized residents on a 17-bed nursing home wing. PMID- 12534871 TI - Guillain-Barre syndrome and hyponatraemia. AB - BACKGROUND: Hyponatraemia is a recognized association of the Guillain-Barre syndrome (GBS) and is also known to occur after the administration of intravenous infusion of gamma globulin (IVIG), a treatment often used in management of GBS. AIMS: To document serum sodium concentration in GBS patients before, during and after management procedures (including IVIG) positive pressure ventilation (PPV) and the use of medications capable of causing a low serum sodium. To consider whether pseudohyponatraemia might be a contributory factor. To assess whether hyponatraemia is a predictor of poor outcome. METHODS: Clinical record audit of 84 patients with GBS admitted to Christchurch Hospital, New Zealand, over a 10 year period. RESULTS: Serum sodium concentration was significantly low (<133 mmol/L) in 26 of 84 (31%) patients. In 12 of these cases the hyponatraemia developed during or after IVIG, suggesting that pseudo-hyponatraemia was a contributing factor. Six (7%) significantly hyponatraemic patients died, but no eunatraemic or slightly hyponatraemic patient died (P = 0.001). Six of 38 patients aged > or =50 years died, whereas none of the 46 patients aged <50 years died (P = 0.007). Five of 19 ventilated patients died but only one of 65 non ventilated died (P = 0.002). CONCLUSIONS: Significant hyponatraemia occurred in 26 (31%) of the patients, but in 12 of these it appeared likely that pseudohyponatraemia caused by IVIG was a contributing factor. Hyponatraemic patients have a poorer prognosis than eunatraemic patients, however it is difficult to separate this factor from other poor prognostic factors (older age and PPV). PMID- 12534872 TI - Evaluation of dosage adjustment in patients with renal impairment. AB - AIMS: To determine whether appropriate dosage adjustments are made in patients with significant renal impairment for drugs with a high fractional renal clearance. METHODS: Evaluation of dosage adjustment was performed in patients who were admitted to a 480-bed metropolitan hospital (Princess Alexandra Hospital, Brisbane, Australia) with an estimated creatinine clearance of < or =40 mL/min. All drugs had a high fractional renal excretion. A prescribed dose within 30% of the calculated dose was considered appropriate. RESULTS: Doses were found to be inappropriately high in 111 (44.8%) of 248 admission prescriptions of the targeted drugs. Doses were appropriately reduced in hospital in 26 patients (23.4%). Seventy-three (29.3%) prescriptions were continued with excessive doses. Only 34 prescriptions for the target drugs were initiated in hospital, of which 88.2% were appropriately dosed. CONCLUSIONS: A significant percentage of patients with renal impairment are admitted to hospital on inappropriately high doses of drugs, with a high fractional renal excretion and low therapeutic index. Doses are appropriately reduced in hospital in some patients but there is still room for improvement [corrected]. PMID- 12534873 TI - What is the prognosis of mild normocytic anaemia in older people? AB - BACKGROUND: There is reduced survival for people with all types of anaemia and it has been shown that there is a low yield from investigations for older people with normocytic anaemia. However, the longer-term outcomes of people with mild normocytic anaemia are not known. AIMS: To determine the 5-year prognosis of older people with mild normocytic anaemia, compared with non-anaemic controls. METHODS: This was a prospective cohort study, with some retrospective data collection, comparing 52 people aged > or =65 years (haemoglobin 100-119 g/L; mean cell volume 80-99 fL) without an obvious cause at presentation with 52 non anaemic controls. The incidence of death, new malignancies and significant illnesses was compared between the two groups, 5 years later. RESULTS: Anaemic patients were less likely to be living at home at baseline. Although survival was lower for the anaemic group at all follow-up times, these differences were not significantly different. Over 5 years, there were no significant differences between groups in the types of problems that developed. CONCLUSIONS: A mild normocytic anaemia in older people, in the absence of an obvious cause, is not associated with significant differences in outcomes but is associated with poorer independence and a trend to lower survival. We suggest it may therefore be a marker of frailty in older people. PMID- 12534875 TI - Endoscopic ultrasound: diagnostic and therapeutic applications. AB - Endoscopic ultrasound (EUS) comprises several techniques of performing high frequency ultrasound via an endoscope placed in the gastrointestinal tract (oesophagus, stomach, duodenum and pancreaticobiliary tree and rectum). It has rapidly become an important tool in the investigation of a variety of lumenal disorders as well as locoregional staging of gastrointestinal malignancies. Needle biopsy of peri-intestinal structures, such as lymph nodes and pancreatic masses, can also be performed under real-time ultrasound control. To date, the utilization of this technology in Australia has been limited by cost constraints and a paucity of training opportunities. EUS continues to be a rapidly growing area in clinical gastroenterology. Recent studies continue to define its role, particularly in the loco-regional staging of a variety of malignancies. In addition, new instruments permit tissue sampling and a variety of therapeutic manoeuvres under direct ultrasound guidance.33; 26-32) PMID- 12534874 TI - Utility of a weight-based heparin nomogram for patients with acute coronary syndromes. AB - BACKGROUND: Unfractionated heparin has been pivotal in the management of acute coronary syndromes (ACS), and continues to be used widely despite the emerging role of low molecular weight heparins (LMWH). The apparent superiority of LMWH over unfractionated heparin may, at least partially, reside in its more predictable achievement of therapeutic effect, with high rates of non-therapeutic activated partial thromboplastin time (APTT) results being observed in the intravenous heparin treatment groups. AIM: To evaluate the impact of introduction of a weight-based heparin nomogram developed for use in patients with ACS on frequency of 'therapeutic' APTT results. METHODS: The effectiveness of an existing non-weight-based heparin nomogram in achieving a therapeutic APTT was compared sequentially with that of a weight-based heparin nomogram in 89 and 84 consecutive patients admitted with a diagnosis of ACS. RESULTS: Patients in whom heparin dosage adjustment was weight based rapidly achieved therapeutic APTT. The median time to achieve an APTT within the target range was 8.75 h in the weight based group versus >24 h in the non-weight-based group. Utilization of a weight based nomogram was associated with markedly increased proportions of readings within the therapeutic APTT range at 6 h and at 24 h (51%vs. 26% and 72%vs. 36%, respectively). CONCLUSIONS: The current study confirms the marked superiority of the weight-based heparin regimen for treatment of patients with ACS. The nomogram dramatically facilitated the attainment of therapeutic APTT, and may represent the optimal method for titration of heparin dosage to individual heparin requirements in patients with ACS. PMID- 12534876 TI - Adjuvant therapy for high-risk primary and resected metastatic melanoma. AB - Patients with thick, primary melanoma and regional lymph-node metastases are at moderate to high risk of recurrence and death, despite apparent complete surgical removal. Immune responses can be demonstrated against melanoma and this has prompted the conduct of a number of randomized trials of immunotherapy. Several trials have been completed and show minimal benefit in prolonging survival or recurrence from melanoma. Similarly, a large number of trials has been conducted to test the efficacy of alpha-2-interferon (IFN-alpha2) in therapy. Clear benefit in recurrence-free survival was shown in several trials, however there is a lack of convincing evidence of an effect on overall survival. Several trials of vaccine and IFN-alpha2 therapy are still in progress and their results are awaited with great interest. The use of high-dose IFN-alpha2 therapy remains a contentious subject, however available evidence suggests the standard of care remains good surgical management. PMID- 12534877 TI - 6-thioguanine: a new old drug to procure remission in inflammatory bowel disease. PMID- 12534878 TI - Vitamin D deficiency is common and unrecognized among recently arrived adult immigrants from The Horn of Africa. AB - The prevalence of and risk factors for vitamin D deficiency were examined for recent adult immigrants from East Africa living in Melbourne. Demographic data were collected via a face-to-face questionnaire and risk factors predicting deficiency were examined. Suboptimal levels <25 nmol/L were found in 61 patients (53%; 74% of women; 20% of men). Lower levels were more likely in: (i) patients with a longer duration of residence in Australia, (ii) patients who were mostly covered when outdoors and (iii) women. Routine assessment for vitamin D deficiency should be considered for male and female immigrants from East Africa. PMID- 12534879 TI - Bilateral adrenal enlargement in Burkitt's lymphoma: infiltration or hyperplasia? PMID- 12534880 TI - Benserazide-induced gynaecomastia. PMID- 12534881 TI - Bartonella henselae infective endocarditis in north Queensland. PMID- 12534882 TI - Meningococcal disease: treatable but still terrifying. PMID- 12534883 TI - Methods for comparison of laboratory tests can be improved. PMID- 12534885 TI - Autoimmune myasthenia gravis: place of thymectomy and preferred technique. PMID- 12534886 TI - One journey, many pathways. PMID- 12534901 TI - Evidence-based meta-analysis of pharmacotherapy for benign prostatic hypertrophy. AB - Evidence-based diagnosis and treatment have been proposed recently in various medical fields. Evidence-based diagnostic and therapeutic guidelines for benign prostatic hypertrophy (BPH), one of the most common urological diseases, have been proposed in foreign countries. This paper examines common therapeutic drugs for BPH in Japan from the viewpoint of evidence-based medicine (EBM). The term 'BPH' and drugs indicated for BPH were used as key words to search related articles in the PubMed website. A total of 813 articles extracted as of October 2001 were examined. The articles were ranked in levels ranging from I-V, where Level I indicated a large randomized controlled trial and Level V indicated a non controlled case accumulation study. Among the 813 articles extracted, 132 clinical articles were suitable for evaluation. There were many reliable articles on the effectiveness of alpha-blockers. However, it seemed necessary to examine further how to choose the optimum alpha-blocker for each clinical case and the combination of antiandrogen drugs with alpha-blockers. It was also considered necessary to evaluate the effectiveness of drugs, such as eviprostat and hachimi jio-gan, that are available only in Japan. The present study evaluated the effectiveness of commonly available therapeutic drugs for BPH in Japan from a viewpoint of EBM. PMID- 12534902 TI - Laparoscopic nephroureterectomy for dysplastic kidney in children: an initial experience. AB - BACKGROUND: Laparoscopic nephroureterectomy for dysplastic kidney is now becoming a widely accepted procedure. We report here our initial experience with laparoscopic nephroureterectomy in four girls. METHODS: Between 1993 and 1999, laparoscopic nephroureterectomy was performed in four girls (mean age 5.3 years). Three patients had an ectopic dysplastic kidney with ectopic ureter, and one patient had hydronephrosis with megaureter due to distal ureteral atresia of the upper moiety in a duplicated dysplastic kidney. The transperitoneal approach was used in all cases. RESULTS: Mean operative time was 195 min (range 150-266). Blood loss was minimal. All operations were completed successfully and there were no intraoperative or postoperative complications except for subcutaneous emphysema in one patient (case 4). Postoperative analgesia was used in three patients and administered in the form of diclofenac sodium suppositories 12.5 mg (cases 1 and 2) or acetaminophen suppositories 50 mg (case 3) for 1-2 days. One patient did not require any analgesia (case 4). Oral fluid intake was resumed on the first postoperative day and ambulation began within 1-3 days (mean 1.6, cases 1, 2 and 3) and 6 days (case 4). All children returned to normal activity within 3-6 days of surgery. Mean postoperative hospital stay was 7.3 days. All cases had uneventful courses after discharge. CONCLUSION: Laparoscopic nephroureterectomy can be performed safely, with minimal postoperative pain, excellent cosmetic results and early ambulation. We advocate the use of laparoscopy for the diagnosis and treatment of dysplastic kidney with ectopic ureter. PMID- 12534903 TI - Gamma-knife radiosurgery for brain metastasis of renal cell carcinoma: results in 42 patients. AB - BACKGROUND: The present study provides data from clinical experience with gamma knife radiosurgery (GK) in patients with brain metastasis from renal cell carcinoma (RCC) and shows the value of this less invasive treatment modality. METHODS: Forty-two patients received GK. Twenty of the 42 cases had multiple brain metastases. Extracranial metastases were observed in the lung (38 cases), bone (12 cases), liver (9 cases), lymph node (5 cases) and skin (6 cases). RESULTS: Neurological symptoms seen in 40 patients were rapidly improved after GK in 32 patients (80%). Magnetic resonance imaging (MRI) evaluation after GK in 32 patients showed the disappearance of brain tumor in 9 patients (28%). Complete response was obtained by GK in tumors up to 30 mm in diameter. Repeated GK for newly developed lesions was conducted in 11 patients. Extracranial tumor resection was conducted in 7 cases (lung: 3, skin: 2, liver: 1, adrenal: 1). Chemo-radiotherapy or immunotherapy was effective in 8 cases (lung: 5, liver: 2, bone: 1). The actual one-, two- and three-year survival rates were 44.9%, 16.8%, and 11.2%, respectively. The median survival time was 12.5 months. In univariate analysis, the patients with successfully treated extracranial metastases had significantly better prognosis. In multivariate analysis, the patients with Karnofsky performance scale (KPS) > or = 80%, who were treated by GK more than once and obtained complete response (CR) or partial response (PR) by GK, had significantly better prognosis. CONCLUSION: Gamma-knife radiosurgery for RCC is an effective non-invasive modality of treatment. It offers a high local control rate and an improved quality of life and survival rate. PMID- 12534906 TI - D-xylose absorption after urinary orthotopic bladder replacement: colon neobladder compared with ileal neobladder. AB - BACKGROUND: To evaluate the digestive and absorptive status using the D-xylose test in patients who underwent radical cystectomy and orthotopic bladder replacement either by colon or ileal segment. METHODS: D-xylose serum levels after an oral load, nutritional status, plasma vitamin B12 levels, and acid-base and electrolyte balances were studied in 18 patients with colon neobladder and 12 patients with ileal neobladder. Mean follow-up period was 51 months. Results of both types of bladder replacement and a healthy control group were compared. RESULTS: Although no significant difference in the changes of plasma levels of D xylose after oral load was observed between patients with colon neobladder and healthy controls, plasma levels of D-xylose 90 min after oral load in patients with ileal neobladder were significantly lower than those with colon neobladder. In contrast, there was no significant difference in nutritional status, plasma levels of vitamin B12, and acid-base and electrolyte balances between patients with colon and ileal neobladders. CONCLUSION: Despite acceptable nutritional status, intestinal malabsorption might be present in patients with ileal neobladder, as indicated by the plasma levels of D-xylose, while the colon neobladder group showed no significant differences compared with normal controls. Therefore, absorptive and metabolic status should be carefully monitored after ileal neobladder creation. PMID- 12534907 TI - Extracorporeal shock wave lithotripsy treatment in paraplegic patients with bladder stones. AB - BACKGROUND: Spinal cord injury patients are at increased risk of developing urolithiasis and many will require treatment, most commonly with extracorporeal shock wave lithotripsy (ESWL). METHODS: We examined 20 consecutive spinal cord injury patients (all male) undergoing ESWL for the treatment of bladder stones between April 1992 and May 2000. The mean number of shock waves administered in these 20 patients was 3600 and the shock power was 19 kW/s. All patients were treated without anesthesia. After ESWL, the bladder was irrigated with povidone iodine solution and stone fragments were evacuated through a 22F endoscopic sheath. RESULTS: All patients became stone-free after ESWL. Stone fragments were observed in the urethra of 3 patients, which were removed by urethroscopy. No major complication was observed; however, 5 patients had minor hematuria (which subsided within 24 h) and subfebrile fever (37.2-37.5 degrees C). CONCLUSION: ESWL and transurethral bladder irrigation is a safe, effective and simple modality for the treatment of bladder stones in spinal cord injury patients. PMID- 12534908 TI - Feasibility and usefulness of laparoscopic radical prostatectomy: Kobe University experience. AB - OBJECTIVES: The objective of this study was to present the clinical outcomes of 26 patients who underwent laparoscopic radical prostatectomy at our institution. METHODS: We performed laparoscopic prostatectomy on patients who were clinical stage T1 or T2. The mean age was 70 years old (range: 52-76). The mean level of pre-treatment prostate-specific antigen (PSA) was 8.7 ng/mL (range: 3.3-45). The Gleason score of the needle biopsy was < 7 in 21 patients and > or = 7 in five patients. Clinical stage was T1c in 17 patients, T2a in 6 patients and T2b in 3 patients. Operative techniques followed those of the French groups. Five trocars were introduced into the peritoneal cavity. The vas deferens and seminal vesicles were dissected to reach the posterior wall of the prostate and the retroperitoneal space was dissected around the urinary bladder. Incision of endopelvic fascia and dorsal vein complex (DVC) ligation were performed. The bladder neck and prostate were divided, then the distal urethra was cut. The lateral pedicles of the prostate were cut and the entire prostate was removed. Vesico-urethral anastomosis was performed at eight points. RESULTS: Mean operation time was 7 h 30 min. Mean bleeding volume (including urine volume) was 850 mL (range: 32-3135). All patients underwent autologous blood transfusion. Only one patient required further blood transfusion. Gleason scores of resected specimens were < 7 in 10 patients, and > or = 7 in 16 patients. Pathological stage was T0 in 1 patient, T2a in 6 patients, T2b in 13 patients, T3a in 5 patients and T3b in 1 patient. The PSA value was undetectable in all patients one month after surgery. Ten patients who survived for 6 months after surgery had complete urinary continence without a pad. In 7 of the 12 patients who were potent before surgery, neurovascular bundles were preserved, and 5 of them (71%) achieved complete or incomplete erection 3 months after surgery. However, only one patient (14%) could have sexual intercourse. CONCLUSION: Although longer follow-up is necessary to evaluate this surgical technique, laparoscopic prostatectomy seems to be a reasonable option in the treatment of organ-confined prostate cancer. PMID- 12534909 TI - Hydrogen-potassium ATPase inhibitors induce relaxation on rabbit prostatic strips in vitro. AB - BACKGROUND: To determine the relaxant effect of omeprazole and lansaprazole, hydrogen-potassium (H+-K+) ATPase inhibitors, on rabbit prostatic tissue in vitro. METHODS: Male New Zealand white rabbits were sacrificed and their prostatic tissues were removed. The prostatic stromal strips were mounted in organ baths and relaxation responses were obtained in precontracted tissues with phenylephrine, carbachol and potassium chloride (KCl). Relaxation responses were controlled in the presence of various antagonists to explain the mechanism for relaxation exerted by omeprazole and lansaprazole. RESULTS: Omeprazole and lansaprazole caused similar relaxation responses in the prostatic strips precontracted with phenylephrine, carbachol and KCl. The addition of prostaglandin synthase inhibitor indomethacin, nitric oxide synthase inhibitor L NAME, potassium channel blockers, glibenclamide and tetraethylammonium into the organ baths did not change the relaxations induced by omeprazole and lansaprazole in vitro. CONCLUSION: Omeprazole and lansaprazole cause a relaxation in prostatic stromal tissue precontracted with phenyephrine, carbachol and KC1 in vitro. This relaxant effect is independent of H+-K+ ATPase inhibition. Additionally, cyclooxygenase and nitric oxide pathways do not contribute to this relaxant effect. Further studies are required to determine whether these drugs may have a beneficial effect in the non-operative treatment of benign prostatic hyperplasia. PMID- 12534911 TI - Renal oncocytomatosis in a long-term hemodialysis patient treated by laparoscopic surgery. AB - A 50-year-old female, who had been on maintenance hemodialysis for 22 years, consulted our clinic because of a left renal mass detected incidentally by ultrasonography. Computed tomography (CT) demonstrated a solid hypervascular mass, suggesting a renal cell carcinoma (RCC), in the left atrophic kidney. Left hand-assisted laparoscopic nephrectomy (HALN) was performed. The histopathological diagnosis was renal oncocytomatosis. Renal oncocytomatosis in a long-term hemodialysis patient is extremely rare. We report the first case of renal oncocytomatosis in a long-term hemodialysis patient treated with hand assisted laparoscopic nephrectomy. PMID- 12534912 TI - Kidney autotransplantation in a patient with renal pelvic and bladder carcinomas. AB - A 66-year-old Japanese woman, whose left kidney was non-functioning, was referred to us with a diagnosis of invasive bladder carcinoma and right renal pelvic carcinoma. The latter tumor was clearly packed in the renal calyx and was considered to be of low-grade malignancy. Nephron preserving surgery of the right kidney was performed, followed by radical cystectomy and construction of an ileal conduit. The postoperative course was good, without significant complications. Renal function was restored and a postoperative imaging study showed a well functioning transplanted kidney. At present (20 months after surgery), the patient is doing well and has presented no signs of recurrence of the disease. PMID- 12534914 TI - Renal metastasis of thyroid carcinoma. AB - We report here the sixth known case of metastatic renal tumor of thyroid carcinoma. In 1999, a 37-year-old man was referred to us with a left renal mass found incidentally by ultrasound during an annual check-up. Computed tomography (CT) revealed a well-defined, hyperdense mass 3 cm in diameter in the middle of the left kidney. Left radical nephrectomy was performed under the preoperative diagnosis of renal cell carcinoma. Histologically, the tumor was metastatic of typical papillary thyroid carcinoma. PMID- 12534913 TI - Extensive bilateral renal pelvis, ureter and bladder leukoplakia. AB - We are presenting a case of leukoplakia spread to the bilateral renal pelvis, ureter and bladder in a 52-year-old woman. We believe this case is important because it involves leukoplakia that is more extensive than in any cases previously cited in the literature. No other possible etiologic cause except a heavy smoking habit was defined. Conservative follow-up was preferred to aggressive surgery. PMID- 12534915 TI - Elevated serum estradiol suggesting recurrence of Leydig cell tumor nine years after radical orchiectomy. AB - We report a case of a Leydig cell tumor that showed retroperitoneal lymph node metastasis 9 years after radical orchiectomy. Elevated serum estradiol (E2) suggested recurrence of the Leydig cell tumor. Retroperitoneal lymph node dissection (RPLND) was performed and the lymph node was proved histopathologically to have a metastatic Leydig cell tumor. After RPLND, serum E2 returned to the normal range. This is the first reported case in which changes in the endocrinological findings were useful as likely tumor markers to detect the recurrence of a Leydig cell tumor. PMID- 12534916 TI - Annual changes of the incidence and clinical characteristics of magnesium ammonium phosphate urinary stones. AB - BACKGROUND: Magnesium ammonium phosphate (MAP) urinary stones account for the majority of staghorn stones and frequently cause a non-functioning kidney. In the present study, we examined the annual changes of the number and clinical characteristics of MAP stones. METHODS: The annual incidence of MAP stones was investigated in 2619 patients with urinary stones in whom composition of the stone was analysed at Chiba University Hospital between 1964 and 1999. In addition, the annual number of patients with MAP stones was examined at Funabashi Clinic. In a total of 644 patients with MAP stones, age and sex of the patients, location and size of the MAP stones, urinary cultures and etiological factors were analysed. RESULTS: The number of MAP stones in the lower urinary tract was relatively constant. In contrast, MAP stones in the upper urinary tract had dramatically decreased since 1989, resulting in an increase in the rate of MAP stones in the lower urinary tract. Age distribution of the MAP stone patients ranged from 10 years to > 80 years, with the majority aged 30-60 years. The proportion of larger MAP stones in the upper urinary tract increased. There was no significant difference in prevalence of urine cultures. Among etiological factors for MAP stones, difficulty on urination tended to be common in recent years. CONCLUSION: The number of MAP stones, especially in upper urinary tract, has been decreasing during the last decade. At present, treatment of urinary tract obstruction seems important for the management of MAP stones in lower urinary tract. PMID- 12534918 TI - Usefulness of diagnostic imaging in primary hyperparathyroidism. AB - BACKGROUND: In patients with primary hyperparathyroidism, prevention of urinary stone recurrence can be achieved by surgical removal of the enlarged parathyroid gland. To ensure the efficacy of surgery for primary hyperparathyroidism, preoperative localization of the enlarged gland is important. In the present study, usefulness of diagnostic imaging for localization of the enlarged gland was investigated in primary hyperparathyroidism. METHODS: We retrospectively examined the findings of imaging studies and clinical records in 79 patients (97 glands) who underwent surgical treatment for primary hyperparathyroidism at Chiba University Hospital between 1976 and 2000. The detection rates of accurate localization were investigated for imaging techniques, such as ultrasonography (US), computed tomography (CT), magnetic resonance imaging (MRI), thallium-201 and technetium-99m pertechnetate (Tl-Tc) subtraction scintigraphy and 99mTc methoxyisobutylisonitrile (MIBI) scintigraphy, and analysed in relation to the size and weight of the gland and pathological diagnosis. RESULTS: The detection rates by US, CT, MRI, Tl-Tc subtraction scintigraphy and MIBI scintigraphy were 70%, 67%, 73%, 38% and 78%, respectively. The overall detection rate changed from 50% to 88% before and after 1987. The detection rate of MIBI scintigraphy was superior to Tl-Tc subtraction scintigraphy. CONCLUSION: In primary hyperparathyroidism, improvement of accurate localization of an enlarged parathyroid gland was demonstrated along with recent advances in imaging techniques including MIBI scintigraphy. PMID- 12534920 TI - Cell proliferation, apoptosis, angiogenesis and growth rate of incidentally found renal cell carcinoma. AB - BACKGROUND: Our previous study showed that the growth rate of incidentally found renal cell carcinoma (RCC) varied, and that the initial clinical and pathological features did not predict subsequent growth of the carcinoma. The objective of this study was to determine the relationships between cell proliferation, apoptosis, angiogenesis and the growth rates of these RCC. METHODS: We examined cell proliferation, apoptosis, and angiogenesis in 16 incidentally found cases of RCC. Cell proliferation was assessed by immunohistochemical staining with a Ki-67 antibody. Apoptosis was assessed by the terminal deoxynucleotidyl transferase (TdT) mediated deoxy-UTP biotin nick end labeling (TUNEL) technique. The Ki-67 labeling index (KI) and the apoptotic index (AI) were determined as the ratio of immunohistochemically positive cells per 1000 cancer cells. The KI/AI ratio was also determined. Angiogenesis was evaluated by CD34 immunostaining. Finally, we investigated the correlation between these parameters and the growth rate of primary lesions of incidentally found RCC. RESULTS: The KI ranged from 7 to 73 (median, 20), AI ranged from 6 to 171 (median, 26), and microvessel density (MVD) ranged from 21 to 673 (median, 265) for incidentally found RCC. Ki-67 labeling index, AI and MVD were not closely correlated to each other. Furthermore, these parameters were not associated with growth rates of incidentally found RCC. Only the KI/AI ratio was strongly correlated to the growth rate of incidentally found RCC (r = 0.709; P = 0.0083). CONCLUSION: Our results suggest that the balance between cell proliferation and apoptosis partly determines the growth rate of primary lesions of incidentally found RCC. PMID- 12534921 TI - Clinical outcome of conservative therapy for stage T1, grade 3 transitional cell carcinoma of the bladder. AB - BACKGROUND: The objective of this study was to retrospectively investigate the effectiveness of transurethral resection of bladder tumor (TURBT) and intravesical instillation therapy for stage T1, grade 3 (T1G3) transitional cell carcinoma (TCC) of the urinary bladder. METHODS: Between January 1995 and December 1997, 97 patients with T1G3 TCC of the urinary bladder were treated by TURBT and adjuvant intravesical instillation with bacillus Calmette-Guerin (BCG) or other anticancer agents. The recurrence-free survival rates were evaluated according to several clinicopathological factors. The cases that progressed to muscle invasive disease were also analysed. RESULTS: In this series, the median follow-up period was 25 months (range, 5- 41) after the initial TURBT. Intravesical recurrence was noted in 44 patients (45%), and the 1, 2, and 3 year recurrence-free survival rates were 72%, 58%, and 42%, respectively. Multivariate analyses revealed that the risk of intravesical recurrence was significantly higher for patients who did not receive BCG therapy, irrespective of age, gender, tumor size, multiplicity, pathological stage, concomitant carcinoma in situ, and lymphovascular involvement. Moreover, after a median of 10 months, disease progression occurred in seven patients (7%), of which only one patient was treated by BCG therapy after initial TURBT. CONCLUSION: These findings suggest that intravesical instillation with BCG combined with TURBT is an effective conservative treatment for T1G3 TCC of the bladder. Patients with negative prognostic factors should be treated by BCG rather than other anticancer agents after TURBT. PMID- 12534922 TI - Antegrade extraperitoneal approach to radical cystectomy and ileal neobladder. AB - BACKGROUND: We describe a new operative technique for the surgical treatment of bladder cancer. METHODS: Male patients with invasive bladder cancer were managed by radical cystoprostatectomy using a technically different approach than the conventional method. The main feature of this method includes a small vertical incision between pubis and umbilicus, keeping the abdominal peritoneum closed during the surgery. Reperitonealization is done at completion to isolate the urinary anastomoses from the bowel anastomoses. RESULTS: Seventy-six consecutive patients with bladder cancer underwent this operation. The mean operation time was 4 h 30 min and the mean hospital stay was 11 days. No mortality was seen in the early postoperative period. Only one patient developed serious bowel distension. Two patients developed pneumonia. Wound infection was seen in two patients. Two patients experienced hydronephrosis in the late period of follow up. Four patients developed pelvic lymphocele. There was no evidence of postoperative electrolyte loss in any of the patients. Three patients developed abdominal hernia and this was corrected with surgical treatment. Urodynamic evaluation of 15 patients showed a low capacity reservoir. All patients were continent during the daytime. CONCLUSION: This technique keeps the abdominal peritoneum closed during radical cystoprostatectomy, preventing the patients from complications, such as infection, water and electrolyte imbalances. The technique also decreases the recovery time. PMID- 12534924 TI - Experimental and clinical trial of measuring urinary velocity with the pitot tube and a transrectal ultrasound guided video urodynamic system. AB - BACKGROUND: The pitot tube is a common device to measure flow velocity. If the pitot tube is used as an urodynamic catheter, urinary velocity and urethral pressure may be measured simultaneously. However, to our knowledge, urodynamic studies with the pitot tube have not been reported. We experimentally and clinically evaluated the feasibility of the pitot tube to measure urinary velocity with a transrectal ultrasound guided video urodynamic system. METHODS: We carried out a basal experiment measuring flow velocity in model urethras of 4.5-8.0 mm in inner diameter with a 12-Fr pitot tube. In a clinical trial, 79 patients underwent transrectal ultrasound guided video urodynamic studies with the 12-Fr pitot tube. Urinary velocity was calculated from dynamic pressure (Pd) with the pitot tube formula and the correcting equation according to the results of the basal experiment. RESULTS: Velocity measured by the pitot tube was proportional to the average velocity in model urethras and the coefficients were determined by diameters of model urethras. We obtained a formula to calculate urinary velocity from the basal experiment. The urinary velocity could be obtained in 32 of 79 patients. Qmax was 8.1 +/- 4.3 mL/s (mean +/- SD; range, 18.4-1.3 mL/s), urethral diameter was 7.3 +/- 3.0 mm (mean +/- SD; range, 18.7 4.3 mm) and urinary velocity was 69.4 +/- 43.6 (mean +/- SD; range, 181.3-0 cm/s) at maximum flow rate. The correlation coefficient of Qmax measured by a flowmeter versus Qdv flow rate calculated with urethral diameter and velocity was 0.41 without significant difference. CONCLUSION: The use of the pitot tube as an urodynamic catheter to a transrectal ultrasound-guided video urodynamic system can measure urethral pressure, diameter and urinary velocity simultaneously. However, a thinner pitot tube and further clinical trials are needed to obtain more accurate results. PMID- 12534925 TI - Transabdominal vesical sonography of urethral syndrome and stress incontinence. AB - BACKGROUND: Transabdominal ultrasonography was used to study the bladder neck morphology in women with urethral syndrome or stress urinary incontinence, in order to determine the ultrasonographic findings of these conditions. METHODS: A total of 210 female patients with a normal bladder, asymptomatic trigonitis, urethral syndrome, and stress incontinence were studied. The mucosal thickness around the bladder neck, the length of the anterior base plate of the bladder, and the anteroposterior vesical wall angle (APVA) at the bladder neck were measured on sagittal transabdominal vesical ultrasonograms with the patient in the supine position. RESULTS: Patients with asymptomatic trigonitis or urethral syndrome had thicker mucosa around the bladder neck than the subjects with a normal bladder, and the subjects with stress incontinence had normal mucosa. The APVA was 158 +/- 17 (mean +/- SD) degrees in the subjects with a normal bladder. It was smaller in symptomatic patients and decreased to 109 +/- 10 degrees in those with conservative therapy-resistant incontinence. The anterior edge of the vesical base plate was visible approximately 2 cm from the bladder neck in subjects without incontinence, while it tended to be absent in patients with incontinence and an APVA of less than 126 degrees. CONCLUSION: A small APVA appears to reflect bladder neck descent, while a small APVA without a visible anterior base plate edge may reflect hypotonia of the vesical base plate. Transabdominal vesical ultrasonography with the patient in the supine position provides useful information and can be carried out as a routine examination in female patients with micturition disorders. PMID- 12534926 TI - Urinary oxalic acid excretion differs after oral loading of rats with various oxalate salts. AB - BACKGROUND: To compare urinary oxalate excretion after the oral administration of oxalic acid, disodium oxalate, or calcium oxalate in rats. METHODS: Male Wistar rats were divided into four groups of six rats each and were intravenously hydrated with normal saline, and then were administered normal saline (control group), 10 mg of oxalic acid, equimolar disodium oxalate, or equimolar calcium oxalate via a gastrostomy. Urine specimens were collected just before administration and at hourly intervals up to 5 h afterwards. The urinary oxalate, calcium, magnesium and phosphorus levels were measured. RESULTS: Urinary oxalate excretion peaked at 1-2 h after administration of oxalic acid or equimolar disodium oxalate, while administration of calcium oxalate only caused a small increase of urinary oxalate excretion. Cumulative urinary oxalate excretion during 5 h was 1.69 +/- 0.10 mg (mean +/- SD; 17%), 1.43 +/- 0.13 mg (13%), and 0.22 +/- 0.03 mg (2%) after the administration of oxalic acid, disodium oxalate, and calcium oxalate, respectively. Urinary calcium excretion showed a decrease in the oxalic acid and disodium oxalate groups, while urinary magnesium or phosphorus excretion did not change significantly. CONCLUSION: The upper gastrointestinal tract seems to be the major site of oxalic acid absorption and only free oxalate is absorbed irrespective of whether it is the sodium salt or not. After binding to calcium in the gut, oxalic acid absorption seems to be inhibited in the presence of calcium and this means that calcium oxalate is poorly absorbed (at least in the upper gastrointestinal tract). PMID- 12534927 TI - Erythropoietin-producing renal cell carcinoma in chronic hemodialysis patients: a report of two cases. AB - Erythropoietin (EPO)-producing renal cell carcinomas in two hemodialysis patients are reported. Despite deteriorated kidney function, these patients did not manifest anemia at diagnosis and their elevated serum EPO levels rapidly returned to within the normal range after nephrectomy. Immunohistochemical staining of the resected specimens showed production of erythropoietin in the tumor cells in one case and in the lining cells of the cyst wall in the other case. Renal cell carcinoma could cause an increase of blood hematocrit level in dialysis patients. PMID- 12534928 TI - Advanced renal cell carcinoma in which a combination of IFN-alpha and meloxicam was thought to be effective. AB - An 83-year-old man with left renal cell carcinoma (RCC; pT4N0M0) was treated with postoperative combined subcutaneous injection therapy of alpha interferon (IFN alpha) and IFN gamma-1a (IFN-gamma-1a). Metastasis to the pleura occurred 3 months after surgery. The metastatic lesion grew while the treatment was changed to intramuscular injection of IFN-alpha-2b due to the presence of severe general malaise, which seemed to be caused by IFN-alpha and IFN-gamma therapy. As melosalgia associated with sciatica was also severe, treatment with meloxicam, which is known as a potent cyclooxigenase-2 inhibitor among commercially available non-steroidal anti-inflammatory drugs, was combined, resulting in significant improvement in activity of daily life, 43.2% decrease in the size of the pleural metastasis and complete regression of retroperitoneal residual tumor. PMID- 12534929 TI - Calcium nephrolithiasis and distal tubular acidosis in type 1 glycogen storage disease. AB - A 36-year-old man was admitted to hospital due to right flank pain as a result of ureteral stones. He had been followed up for type 1 glycogen storage disease since the age of 11 years. He had four episodes of spontaneous stone birth during the previous 2 years, and each stone was composed mainly of calcium oxalate. Intravenous pyelography showed right hydronephrosis due to ureteral stones and bilateral multiple renal stones. We carried out transurethral ureterolithotripsy (TUL) on the right ureteral stones. The composition was a mixture of calcium oxalate and calcium phosphate. Laboratory evaluation demonstrated the association of distal renal tubular acidosis (RTA). These observations suggest that hypocitraturia and distal RTA are strongly correlated to recurrence of calcium nephrolithiasis. The patient's serum uric acid and urinary citrate excretion levels normalized after allopurinol and potassium citrate administration. PMID- 12534930 TI - Primary bilateral spermatocele. AB - We report the case of a bilateral, relatively huge, multilocular spermatocele diagnosed at sonography with surgical and pathological correlation. A 45-year-old man presented with bilateral, large inguinoscrotal masses. He had had the masses since his adolescence and their size gradually increased in the past 5 years. Scrotal ultrasonographic examination showed cystic fluid masses that were 65 x 45 x 50 mm in size on the right and 55 x 45 x 40 mm in size on the left side in multilocular spaces. The cystic masses were excised via bilateral inguinoscrotal incision staying near the body of the epididymis. The patient had fathered three children with no fertility problem. PMID- 12534932 TI - The 'oestrogen hypothesis'- where do we stand now? AB - The original 'oestrogen hypothesis' postulated that the apparent increase in human male reproductive developmental disorders (testis cancer, cryptorchidism, hypospadias, low sperm counts) might have occurred because of increased oestrogen exposure of the human foetus/neonate; five potential routes of exposure were considered. This review revisits this hypothesis in the light of the data to have emerged since 1993. It addresses whether there is a secular increasing trend in the listed disorders and highlights the limitations of available data and how these are being addressed. It considers whether new data has emerged to support the suggestion that increased oestrogen exposure could cause these abnormalities and reviews new data on potential routes via which such increased exposure could have occurred. Secular trends: The disorders listed above are now considered to represent a syndrome of disorders (testicular dysgenesis syndrome, TDS) with a common origin in foetal life. Testicular cancer has increased in incidence in Caucasian men worldwide and lifetime risk is 0.3-0.8%. Secular trends in cryptorchidism are unclear but it is by far the commonest (2-4% at birth) congenital abnormality in either sex. Secular trends for hypospadias are not robust, although most studies suggest a progressive increase; registry data probably under-estimates incidence, but based on this data hypospadias is the second most common (0.3-0.7% at birth) congenital malformation. Retrospective analyses of sperm count data show a global downward trend but this is inconclusive - prospective studies using standardized methodology show significant differences between countries and very low sperm counts in the youngest cohort of men. For all disorders, other then testis cancer, standardized prospective studies are the best way forward and are in progress across Europe. Oestrogen effects: Evidence that foetal exposure to oestrogens can induce the above disorders has strengthened. New pathways via which such changes could be induced have been identified, including suppression of testosterone production by the foetal testis, suppression of androgen receptor expression and suppression of insulin-like factor-3 (InsL3) production by foetal Leydig cells. Other evidence suggests that the balance between androgen and oestrogen action may be important in induction of reproductive tract abnormalities. Oestrogen exposure: Although many new environmental oestrogens have been identified, their uniformly weak oestrogenicity excludes the possibility that they could induce the above disorders. However, emerging data implicates various environmental chemicals in being able to alter endogenous levels of androgens (certain phthalates) and oestrogens (polychlorinated biphenyls, polyhalogenated hydrocarbons), and the former have been shown to induce a similar collection of disorders to TDS. Other mechanisms via which increased fetal exposure to pregnancy oestrogens might occur (increasing trend in obesity, dietary changes) are also discussed. PMID- 12534933 TI - Androgen insensitivity and male infertility. AB - Abnormal human spermatogenesis can be caused by defects in androgen action because of androgen insensitivity. A variety of mutations have been described in the human androgen receptor gene associated with male infertility. These can be attributed to two molecular mechanisms. First, point mutations in the androgen receptor gene cause alterations in the amino acid sequence and, hence, lead to apparently slight changes in the androgen receptor effector mechanisms and mild androgen insensitivity. Secondly, variations in the polymorphic poly glutamine segment within the N-terminal end of the androgen receptor have been ascribed to correlate with fertility aspects possibly because of modifications of transcriptional regulatory mechanisms. It has been postulated that longer poly glutamine segments are associated with decreased sperm counts. However, the molecular mechanisms that lead to inhibition of spermatogenesis because of a mutated androgen receptor are poorly understood and will need more focus in the future. PMID- 12534934 TI - Androgen receptor and prostate cancer invasion. AB - Evidence indicates that androgen-sensitive prostate cancer cells have a lower malignant potential. We previously demonstrated that expression of androgen receptor (AR) by transfection of the androgen-independent prostate cancer cell line PC3 decreases invasion and adhesion of these cells through modulation of alpha6beta4 expression. Treatment with the androgen further reduced adhesion and invasion of the cells without, however, modifying alpha6beta4. Here we investigated whether the androgen has a direct effect on alpha6beta4-EGF receptor (EGFR) interaction and signalling leading to invasion of these cells. Immunoconfocal microscopy demonstrated that in control cells (PC3-Neo), alpha6beta4 and EGFR colocalize and redistribute in response to epidermal growth factor (EGF). In PC3-AR cells colocalization and redistribution between the two molecules was reduced and abolished by pre-treatment with R1881. Co immunoprecipitation studies demonstrated that tyrosine phosphorylation of beta4 in response to EGF was reduced in PC3-AR cells compared to PC3-Neo. Immunoconfocal and co-immunoprecipitation studies demonstrated colocalization at membrane level and co-immunoprecipitation of EGFR and AR, indicating an interaction between the two proteins. PI3K activity, a key signalling pathway for invasion of these cells, was decreased in PC3-AR cells in response to EGF and further reduced by treatment with R1881. EGFR internalization was strongly reduced in PC3-AR compared with PC3-Neo cells and was reduced by treatment with R1881. In conclusion, the expression of AR by transfection in PC3 cells confers a less malignant phenotype by interfering with EGFR--alpha6beta4 interaction and signalling leading to invasion through a mechanism involving an interaction between the classic AR and EGFR. PMID- 12534935 TI - Effect of neonatal treatment of rats with potent or weak (environmental) oestrogens, or with a GnRH antagonist, on Leydig cell development and function through puberty into adulthood. AB - This study addressed whether reduced Sertoli cell number or manipulation of the neonatal hormone environment has an influence on final Leydig cell number per testis in the rat, by applying neonatal treatments known to affect these parameters, namely administration of a GnRH antagonist (GnRHa) or diethylstilboestrol (DES, in doses of 10, 1 or 0.1 microg per injection). The effect of treatment with either of two 'environmental oestrogens', bisphenol-A (Bis-A) or octylphenol (OP), was also evaluated. Leydig (3beta-hydroxysteroid dehydrogenase immunopositive) cell development and function (plasma testosterone levels) were studied through puberty into adulthood. Treatment with GnRHa impaired testis growth, Leydig cell (nuclear) volume per testis and testosterone levels during puberty, when compared with controls, but final Leydig cell volume/number in adulthood was comparable with controls. As adult testis weight was reduced by 45% in GnRHa-treated rats, the percentage Leydig cell volume per testis was approximately double (p < 0.01) that in controls, and also at day 35. Testosterone levels in adulthood in GnRHa-treated rats were lower (p < 0.01) than in controls but were within the lower end of the normal range. Treatment with DES caused largely dose-dependent suppression of testis growth, Leydig cell (nuclear) volume per testis and testosterone levels up to day 35. Although by adulthood, Leydig cell volume/number per testis was comparable with controls in DES-treated rats, testosterone levels remained grossly subnormal. Neonatal treatment with either Bis-A or OP had little consistent effect on any of the parameters studied except that both treatments significantly elevated testosterone levels on day 18, as did treatment with DES-0.1 microg. The present findings are interpreted in the context of what is known about the hormonal regulation of Leydig cell development. These lead to the conclusion that final Leydig cell number per testis is not determined by the number of Sertoli cells per testis and appears not to be influenced in any major way by gonadotrophins, androgens or oestrogens in the first 2 weeks of postnatal life. This implies that adult Leydig cell number may be determined prior to birth. PMID- 12534936 TI - Expression and regulation of the prointerleukin-1alpha processing enzymes calpain I and II in the rat testis. AB - Interleukin-1alpha (IL-1alpha) is constitutively expressed in an age- and stage dependent manner by rat Sertoli cells. However, the mechanism of regulation of IL 1alpha is unclear in testis. We studied this regulation at the level of the enzyme calpain, a potential regulator that cleaves 32 kDa proIL-1alpha to produce mature 17 kDa IL-1alpha. Both calpain I and II were found to cleave recombinant rat testis 32proIL-1alpha in vitro. A temporary age-related increase in messenger RNA (mRNA) levels of calpain I was found in testis of 20- and 25-day-old rats, coinciding with important events of spermatogenesis and a gradual increase in IL 1alpha, while calpain II expression was constant. In response to lipopolysaccharide (LPS), calpain I protein levels were down-regulated in the seminiferous tubules, while calpain II was less affected. By contrast, the liver after LPS treatment showed up-regulated calpain I and II immunoreactive protein and reverse transcriptase chain reaction (RT-PCR) signal. Depleting Leydig cells by ethane 1,2-dimethane sulphonate treatment resulted in down-regulated calpain I mRNA and protein expression, whereas calpain II remained unchanged. In summary, there is a differential expression of calpain I and II under pathological conditions induced either by endotoxin stimuli or Leydig cell depletion, which may produce a differential effect on IL-1alpha processing. PMID- 12534937 TI - Absence of association of androgen receptor trinucleotide expansion and poor semen quality. AB - This study investigated the relationship between variation in the polymorphic CAG trinucleotide repeat (TNR) region of the human androgen receptor (AR) gene and semen quality in a Caucasian sample population. These men were patients attending the New Zealand Centre for Reproductive Medicine in Christchurch. The AR TNR region was amplified by polymerase chain reaction and then DNA sequenced to determine exact numbers of CAG repeats for each sample. In addition, the samples were screened for microdeletions within the AZFc region of the Y-chromosome. A total of 105 men with poor semen quality were compared with a group of 93 men with normal semen quality. Men with poor semen quality had similar CAG repeat number to men with normal semen quality (21.46 +/- 0.30 vs. 20.99 +/- 0.28, p = 0.126). Y-chromosome microdeletions were only detected in men with suboptimal semen parameters (7.4%). However, the presence of a deletion was not related to CAG repeat number. The CAG repeat number in the men with normal semen quality in the present study is similar to the Australian and German samples, but lower than those reported for the Swedes, Dutch and Danes. These results are contrary to the hypothesis that higher CAG repeats are associated with infertility in men, but strongly suggest that different populations may show different numbers of CAG repeats in addition to racial variation reported in previous studies. PMID- 12534938 TI - Differential expression of succinyl CoA transferase (SCOT) genes in somatic and germline cells of the mouse testis. AB - Succinyl CoA:3-oxo acid CoA transferase (SCOT/OXCT; EC 2.8.3.5) is a key mitochondrial enzyme in the metabolism of ketone bodies in various organs (but not in the liver). We identified a cDNA clone of the testicular germ cell specific succinyl CoA transferase isozyme (SCOT-t). We then isolated a mouse orthologue of the SCOT/OXCT cDNA (SCOT-s) and determined the expression of the two types of SCOT in the testis. The mRNAs of scot-s and scot-t were expressed exclusively in testicular somatic cells (i.e. Leydig and Sertoli cells) and germ cells, respectively. SCOT enzymatic activities were assayed in Leydig cell (SCOT s) and sperm (SCOT-t) fractions. The SCOT activity in sperm was 2.5-fold higher than that in Leydig cells. We conclude that germ cells and somatic cells differentially express the SCOT enzymes and that the SCOT activity of sperm caused exclusively by SCOT-t should play an important role in sperm activity. PMID- 12534939 TI - Comparison of sibling oocyte outcomes after intracytoplasmic sperm injection and in vitro fertilization in severe teratozoospermic patients in the first cycle. AB - To follow up the outcome of sibling oocytes subjected to both conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in the first cycles of severe teratozoospermic patients with normal sperm morphology (NSM)or=6 cell embryos on day 3 and blastocyst formation on days 5 and 6, did not differ significantly between the two groups. There was a trend towards a high pregnancy rate cycle in mixed ICSI/IVF embryo transfer (ET) (49.1%). In conclusion, this study showed that in couples with only severe teratozoospermia, there was a benefit in subjecting sibling oocytes to both IVF and ICSI in the first cycle because 24 (28.2%) cycles of total fertilization failure were avoided. Furthermore, despite initially significant higher fertilization rates in ICSI than IVF oocytes, subsequent rates of development from >or=6 cells up to blastocyst stage were the same suggesting that ICSI should be used with caution, as after day 3, ICSI-derived embryo development was compromised compared with IVF. PMID- 12534940 TI - Paucity of functional CTL epitopes in the E7 oncoprotein of cervical cancer associated human papillomavirus type 16. AB - Many specific antiviral and antitumour immune responses have been attributed to the protective effects of antigen-specific CD8+ cytotoxic T lymphocytes (CTL). Recognition of virus infected or tumour cells by CTL requires presentation of at least one peptide epitope from a virus or tumour-specific antigen by the relevant MHC Class I molecule. Viral genes with mutations which remove CTL epitopes may thus be favoured for survival. Human cervical cancers are caused by papillomavirus infection, and these cancers consistently express the E7 protein of the oncogenic papillomavirus. We therefore investigated the MHC Class I restricted T cell epitopes of the human papillomavirus type 16 E7 oncoprotein using mice of five different genetic backgrounds, and an IFN-gamma ELISPOT assay, to determine the frequency with which MHC Class I epitopes might be expected in this small oncoprotein (98 amino acids). No MHC Class I restricted responses were detected in E7 immunized BALB/c (H-2d), CBA/CaH (H-2 k), FVB/N (H-2q) or A2KbH2b human HLA2.1 transgenic mice. In C57BL/6 J (H-2b) mice, a previously identified single antigenic epitope was detected. Therefore, we conclude that there is a paucity of MHC Class I restricted T cell epitopes in HPV16 E7 protein because of its small size. This might be advantageous to the virus. Furthermore here we present a quick and easy method to exhaustively determine CD8 T cell epitopes in proteins using a unique set of overlapping 8, 9 and 10 mer synthetic peptides. PMID- 12534941 TI - IL-4 responsive CD4+ T cells specific for myelin basic protein: IL-2 confers a prolonged postactivation refractory phase. AB - This study compared myelin basic protein-specific T cells from Lewis rats that were derived in the presence of either rat IL-4 or IL-2. Interleukin-4 was a maintenance factor that enabled derivation of long-term T cell lines. When activated, IL-4 dependent lines were lacking in IL-2 production capacity but maintained high levels of responsiveness to IL-2 and recognized IL-2 as a dominant growth factor. Activated IL-4 dependent T cells rapidly reverted to a quiescent phenotype in the presence of IL-4 and rapidly regained myelin basic protein reactivity. In contrast, activated IL-2 dependent T cells that were propagated in IL-2 had a more persistent blastogenic phenotype and a prolonged refractory phase. Interleukin-4 dependent lines that were propagated in IL-2 up regulated the capacity to produce IL-2 and also acquired prolonged postactivation refractoriness. Thus, IL-2 was a dominant growth factor that conferred prolonged activation-dependent non-responsiveness. The coupling of dominant growth factor activity with prolonged postactivation refractoriness may be associated with the requisite role of IL-2 in homeostatic self-tolerance. PMID- 12534942 TI - Gates, GAVI, the glorious global funds and more: all you ever wanted to know. AB - Global immunization programmes have achieved some remarkable successes. In 1977, Frank Fenner's Commission declared smallpox to have been eradicated by an 11-year long intensive campaign. The Expanded Programme on Immunization encompassed six important childhood vaccines and reached over three-quarters of the world's children. Polio eradication has gone remarkably well, with only 10 out of 200 countries reporting residual cases. But amidst all the good news, there is also bad news. Coverage is variable; infrastructure is crumbling; and newer vaccines are not being incorporated in many country programmes. The Bill and Melinda Gates Foundation has introduced a new dynamic here. From their initial gift of $100 million in December 1998, their commitment to date is US$1.5 billion - and rising. At the centre is a Global Children's Vaccine Fund which permitted the launch, in January 2000, of the Global Alliance for Vaccines and Immunization. This is targeted to the 74 poorest countries of the world and is designed to improve vaccination infrastructure, to purchase newer vaccines and to support research and development. Even before we know how successful this programme will be, it has had its imitators. The Global Fund to Fight AIDS, TB and Malaria borrowed many concepts from GAVI. The Global Alliance for Improved Nutrition announced in May 2002 does so as well, and is heavily supported by Gates. Highly effective parasite control programmes antedate all this but will be much strengthened. However, we still face a sizeable budgetary gap both for research and for bringing the best advances to all people who need them. PMID- 12534943 TI - Mucosal immunity: some historical perspective on host-pathogen interactions and implications for mucosal vaccines. PMID- 12534944 TI - Improving vaccines against tuberculosis. AB - Tuberculosis remains a major cause of mortality and physical and economic deprivation worldwide. There have been significant recent advances in our understanding of the Mycobacterium tuberculosis genome, mycobacterial genetics and the host determinants of protective immunity. Nevertheless, the challenge is to harness this information to develop a more effective vaccine than BCG, the attenuated strain of Mycobacterium bovis derived by Calmette and Guerin nearly 90 years ago. Some of the limitations of BCG include the waning of the protective immunity with time, reduced effectiveness against pulmonary tuberculosis compared to disseminated disease, and the problems of a live vaccine in immuno-compromised subjects. Two broad approaches to vaccine development are being pursued. New live vaccines include either attenuated strains of Mycobacterium tuberculosis produced by random mutagenesis or targeted deletion of putative virulence factors, or by genetic manipulation of BCG to express new antigens or cytokines. The second approach utilizes non-viable subunit vaccines to deliver immunodominant mycobacterial antigens. Both protein and DNA vaccines induce partial protection against experimental tuberculosis infection in mice, however, their efficacy has generally been equivalent to or less than that of BCG. The comparative effects of cytokine adjuvants and vaccines targeting antigen presenting cells on enhancing protection will be discussed. Coimmunization with plasmid interleukin-12 and a DNA vaccine expressing Antigen 85B, a major secreted protein, was as protective as BCG. The combination of priming with DNA-85B and boosting with BCG was superior to BCG alone. Therefore it is possible to achieve a greater level of protection against tuberculosis than with BCG, and this highlights the potential for new tuberculosis vaccines in humans. PMID- 12534945 TI - Bacterial otitis media: current vaccine development strategies. AB - Otitis media is the most common reason for children less than 5 years of age to visit a medical practitioner. Whilst the disease rarely results in death, there is significant associated morbidity. The most common complication is loss of hearing at a critical stage of the development of speech, language and cognitive abilities in children. The cause and pathogenesis of otitis media is multifactorial. Among the contributing factors, the single most important are viral and bacterial infections. Infection with respiratory syncytial virus, influenza viruses, para-influenza viruses, enteroviruses and adenovirus are most commonly associated with acute and chronic otitis media. Streptococcus pneumoniae, non-typeable Haemophilus influenzae and Moraxella catarrhalis are the most commonly isolated bacteria from the middle ears of children with otitis media. Treatment of otitis media has largely relied on the administration of antimicrobials and surgical intervention. However, attention has recently focused on the development of a vaccine. For a vaccine to be effective against bacterial otitis media, it must, at the very least, contain antigens that induce a protective immune response in the middle ear against the three most common infecting bacteria. Whilst over the past decade there has been significant progress in the development of vaccines against invasive S. pneumoniae disease, these vaccines are less efficacious for otitis media. The search for candidate vaccine antigens for non-typeable H. influenzae are well advanced whilst less progress has been made for M. catarrhalis. No human studies have been conducted for non-typeable H. influenzae or M. catarrhalis and the concept of a tribacterial vaccine remains to be tested in animal models. Only when vaccine antigens are determined and an understanding of the immune responses induced in the middle ear by infection and immunization is gained will the formulation of a tribacterial vaccine against otitis media be possible. PMID- 12534946 TI - Immune responses to hepatitis B surface antigen following epidermal powder immunization. AB - Langerhans cells in the epidermis of skin are potent antigen-presenting cells that trigger the immune system to respond to invading microorganisms. We have previously shown that epidermal powder immunization with a powdered inactivated influenza virus vaccine, by targeting the Langerhans cell-rich epidermis, was more efficacious than deeper tissue injection using a needle and syringe. We now report enhanced humoral and cellular immune responses to recombinant hepatitis B surface antigen following epidermal powder immunization. We observed that epidermal powder immunization with unadjuvanted hepatitis B surface antigen elicited an antibody titre equivalent to that induced by the alum-adjuvanted vaccine delivered by intramuscular injection, suggesting that epidermal powder immunization can overcome the need for adjuvantation. We demonstrated that synthetic CpG oligonucleotides (CpG DNA) could be coformulated with hepatitis B surface antigen and delivered by epidermal powder immunization to further augment the antibody response and modulate T helper cell activities. Epidermal powder immunization of hepatitis B surface antigen formulated with CpG DNA formulations resulted in 1.5-2.0 logs higher IgG antibody titres than alum-adjuvanted commercial vaccines administered by intramuscular injection. Formulation of hepatitis B surface antigen with CpG DNA elicited an augmented IgG2a antibody response and increased frequency of IFN-gamma secreting cells. In addition, CpG DNA was found to activate epidermal Langerhans cells and stimulate the production of TNF-alpha and IL-12 cytokines by epidermal cells, explaining its strong adjuvant activity following epidermal powder immunization. These results show that epidermal powder immunization is a safe and effective method to deliver hepatitis B surface antigen and the addition of new adjuvants, such as CpG DNA, may further enhance the efficacy of this vaccine. PMID- 12534947 TI - CPG ODN allows lower dose of antigen against hepatitis B surface antigen in BALB/c mice. AB - We have evaluated alum, immunostimulatory cytosine guanine dinucleotide containing oligodeoxy-nucleotides (CPG ODN), and an alum/CPG ODN combination as adjuvants with hepatitis B surface antigen, to compare their potential to allow lower doses of antigen to be used for induction of humoral responses. BALB/c mice were immunized by intramuscular injection with 0.01, 0.1 or 1.0 micro g recombinant hepatitis B surface antigen without adjuvant or with alum and/or CPG ODN added. When given without adjuvant or with alum, each 10-fold decrease in amount of antigen resulted in a similarly reduced titre of antibody against hepatitis B surface antigen. In contrast, CPG ODN, on its own or combined with alum, allowed high anti-hepatitis B surface antigen titres even with a 1000-fold reduction in amount of antigen. These findings may have important immunological and economical consequences for vaccine development. PMID- 12534948 TI - Immune responses in hepatitis C virus infection: the role of dendritic cells. AB - Cellular immune responses are critical for the clearance of hepatitis C virus. Persistent infection results from a narrow and weak cellular immune response, in direct contrast to the broad, strong response associated with viral clearance in acute infection. The presence of dendritic cells in the liver facilitates presentation of viral antigens to both CD4+ and CD8+ T cell populations. Exploiting the potent antigen presentation capability of dendritic cells for immunotherapy of chronic hepatitis C is attractive; however, infection or transfection of segments or the entire hepatitis C virus genome appears to impair the allostimulation capacity of dendritic cells. If dendritic cell immunotherapy for hepatitis C virus infection is to become a reality, the mechanism behind the defective allostimulatory capacity needs to be deciphered. PMID- 12534949 TI - Maturation of dendritic cells with lipopeptides that represent vaccine candidates for hepatitis C virus. AB - The ability of antigens to elicit immune responses depends upon their initial recognition, uptake, processing and presentation by dendritic cells. This fact has been recognized by many workers and dendritic cells are now regarded as natural 'adjuvants' in the business of vaccine design. One way of persuading dendritic cells to become interested in foreign material is to decorate it with lipid moieties found in bacteria. This approach has been used in the context of synthetic peptide-based immunogens and depending on the nature of the epitopes included, can provide highly immunogenic structures capable of eliciting antibody or cytotoxic T cell responses. In this paper we describe the results of experiments in which the stimulatory effects of peptide-based vaccine candidates on human dendritic cells are examined. Our findings indicate that lipidated structures comprising vaccine target sequences of viral origin coupled to the synthetic lipid groups of bacteria are able to induce the maturation of dendritic cells, as measured by the expression of cell surface MHC class II molecules. PMID- 12534950 TI - Generation of CTL responses using Kunjin replicon RNA. AB - The Kunjin replicon was used to express a polytope that consisted of seven hepatitis C virus cytotoxic T lymphocyte epitopes and one influenza cytotoxic T lymphocyte epitope for vaccination studies. The self-replicating nature of, and expression from, the ribonucleic acid was confirmed in vitro. Initial vaccinations with one dose of Kun-Poly ribonucleic acid showed that an influenza specific cytotoxic T lymphocyte response was elicited more efficiently by intradermal inoculation compared with intramuscular delivery. Two micrograms of ribonucleic acid delivered in the ear pinnae of mice was sufficient to elicit a detectable cytotoxic T lymphocyte response 10 days post-vaccination. Further vaccination studies showed that four of the seven hepatitis C virus cytotoxic T lymphocyte epitopes were able to elicit weak cytotoxic T lymphocyte responses whereas the influenza epitope was able to elicit strong, specific cytotoxic T lymphocyte responses following three doses of Kun-Poly ribonucleic acid. These studies vindicate the use of the Kunjin replicon as a vector to deliver encoded proteins for the development of cell-mediated immune responses. PMID- 12534951 TI - Aspects of cancer immunotherapy. AB - Cancer immunotherapy has traditionally undergone a 'revolution' every decade, from the use of Bacille Calmette-Guerin by scarification in the 1970s, to interleukin-2 therapies in the 1980s, and monoclonal antibody treatments in the early 1990s. Usually the early reports on the use of such agents were encouraging, but when more patients were studied in multiple centres, the initial promising results could not be confirmed. Now in a new century, we have more reagents and methods available than ever before - indeed, with such a plethora of reagents it is difficult to envisage them being fully and appropriately tested within the next decade, by which time there will be even more reagents to test. However, there have been three major advances which should lead to substantial progress in cancer immunotherapy: (1) the widespread use of genetic engineering, enabling identification of candidate vaccine proteins and manipulation of their sequences; (2) the production of antigens, antibodies and cytokines in large amounts by recombinant technologies, and (3) an understanding of the mode of presentation of peptides by major histocompatibility complex Class I and Class II molecules and their recognition by T cells. Despite these advances, there are major problems facing cancer immunotherapy, such as the ability of tumours to mutate and evade the immune system and the difficulty of precisely defining the interactions of effector cells in mediating 'rejection' or destruction of a tumour. There are clearly immunological similarities with diseases such as malaria and schistosomiasis, where the invading foreign organisms can use a variety of strategies to resist an elicited immune response. The failure to find a suitable vaccine for these diseases must lead to some pessimism for the development of immunotherapy for an autologous tumour. However, there are promising studies now in progress which should give an indication of the most important directions to follow. This review provides a commentary on aspects of cancer immunotherapy and in particular will deal with: (1) the selection of antigens as vaccine components; (2) the modes of presentation of antigens, particularly by major histocompatibility complex Class I molecules; and (3) new modes of delivery of vaccine immunogens. PMID- 12534952 TI - Hypoallergenic derivatives of major grass pollen allergens for allergy vaccination. AB - Grass pollen-induced hay-fever and allergic asthma represent a major health problem in industrialized countries. Whereas the symptoms of these allergic conditions can be controlled by pharmacotherapy, specific immunotherapy vaccination is the only causative approach towards the treatment of these type 1 allergies. Specific immunotherapy is based on administration of increasing amounts of the disease-causing allergens in the form of allergen-containing extracts. However, the extracts used for immunotherapy consist of allergenic and non-allergenic components and may induce severe anaphylactic side-effects upon therapeutic administration. With recent developments in molecular biology of pollen allergens it has become feasible to produce modified hypoallergenic derivatives of recombinant allergens with abrogated or greatly reduced likelihood of anaphylactic side-effects as compared to extract-based treatments. We have demonstrated this concept through reducing the anaphylactic potential of major rye grass pollen allergens by introducing a few point mutations which leave the overall structural fold of the molecule unaltered. These modified forms are expected to make allergen-specific immunotherapy more widely used in the future. PMID- 12534953 TI - Modelling the 3-year risk of myocardial infarction among participants in the Data Collection on Adverse Events of Anti-HIV Drugs (DAD) study. AB - OBJECTIVES: To estimate the 3-year risk of myocardial infarction (MI) among participants in the Data Collection on Adverse Events of Anti-HIV Drugs (DAD) study. METHODS: Conventional cardiovascular risk equations were applied to baseline data from the DAD study to estimate the 3-year risk of MI. Best estimates were obtained by simply applying the risk equations, with upper and lower limits based on worst case and optimistic case scenarios. Three-year risks of AIDS or death were also estimated based on a prognostic scoring system for patients receiving antiretroviral (ARV) treatment, and on estimated AIDS rates in untreated people with HIV for those patients not on ARVs or if they were to cease ARVs. RESULTS: Analyses were based on 17 600 patients (24.3% female) recruited into the DAD study with baseline data and no previous MI. The overall 3-year risk of MI was estimated to be 0.72% (lower limit 0.35, upper limit 1.12%), corresponding to a total predicted 127 (65-197) MIs over a 3-year follow-up period. The risk was much greater for men than women (0.92% vs. 0.07%), with only three (2-8) MIs predicted in women. The 3-year risk of MI was estimated to increase from 0.30% (0.20-0.38%) in ARV naive patients to 1.07% (0.43-1.77%) in patients receiving ARVs from all three drug classes. The estimated 3-year risk of AIDS or death was in the range 6.2% to 11.1% in patients receiving ARVs if they continued treatment, and 22.5% to 29.4% if they ceased ARVs. DISCUSSION: These models suggest that although the increase in relative risk of MI as a result of ARV treatment may be as high as threefold in a worst case scenario, the absolute risk is modest with a best estimate of 3-year risk less than or equal to 1% in all groups of patients, and is outweighed by the benefits of ARV treatment in terms of reduced risk of AIDS and death in most patients. As estimates are based on models not validated for people receiving ARV drugs, all estimates should be interpreted cautiously. PMID- 12534954 TI - Findings from the first national clinical audit of treatment for people with HIV. AB - OBJECTIVES: We reviewed the impact of and assessed adherence to British HIV Association (BHIVA) guidelines in routine clinical practice. Feedback has been provided to clinical centres to facilitate any necessary change. METHODS: We used a questionnaire to gauge clinicians' views on the guidelines and availability of antiretroviral therapy (ART) drugs and specialized tests. A case note review of 2044 patients was conducted to assess adherence to guideline recommendations plus patterns of use of HIV resistance testing. RESULTS: Most clinicians (74.1%) report that BHIVA guidelines have influenced care at their centres. A significant minority report problems with access to specialized tests. Most patients who started ART did so at CD4 counts lower than guidelines recommend but in most cases this reflected the CD4 count at diagnosis of HIV. Of patients on ART, an overwhelming majority (97.6%) were receiving three or more drugs. Of those on three or more drugs, 58.9% had latest viral load (VL) below 50 HIV-1 RNA copies/mL and a further 18.1% below 500 copies/mL. Only 19.3% of patients had been tested for HIV resistance, of whom more than half showed resistance to more than one class of drugs. CONCLUSIONS: This clinical audit provides encouraging evidence of the quality of care offered to people with diagnosed HIV in the UK. However late diagnosis means most people start ART at a more advanced stage than guidelines recommend. PMID- 12534955 TI - Prevalence, risk factors and outcome of hyperlactataemia in HIV-infected patients. AB - OBJECTIVE: We describe the prevalence, risk factors and outcome of hyperlactataemia (HL) in a cohort of 140 HIV-infected patients. PATIENTS AND METHODS: Patients were enrolled consecutively within a 3-month period (July to September 1999) and followed until 31 October 2000. One hundred and forty HIV infected patients had venous plasma lactate levels measured. HL was defined at baseline by two consecutive lactate levels > 2.1 mmol/L (upper limit of normal). We compared baseline demographic characteristics, immuno-virological parameters, antiretroviral therapy and outcome between patients with HL (cases) or without HL (controls). We described the clinical features of patients with HL. RESULTS: Among 129 patients included in the analysis, HL was found in 11 patients (8.5%), all of whom were receiving nucleoside reverse transcriptase inhibitors (NRTIs). Cases were more likely than controls to receive didanosine or stavudine (82% vs. 19%, P= 2.7 x 10(-6) and 82% vs. 48%, P= 0.03, respectively). Only 4/11 cases (36%) had symptoms consistent with HL. After a median follow-up of 15 months, lactate level returned to normal in all three patients who discontinued NRTIs, but in only 2/8 patients who did not (P = 0.06). Only one case experienced lactic acidosis and died during follow-up. Mortality rate was similar in cases and controls. CONCLUSION: HL is associated with NRTI use, in particular didanosine and stavudine, and discontinuation of NRTIs seems to be associated with rapid resolution of HL. Lactic acidosis remains rare and the long-term outcome of patients with HL does not seem to be poorer than controls. PMID- 12534956 TI - Patterns of antiretroviral use in the United States of America: analysis of three observational databases. AB - OBJECTIVE: To characterize patterns of antiretroviral use in HIV-infected patients and explore variation by patient characteristics and disease stage. METHODS: Three large patient databases recording information derived from routine clinical attendance were analyzed: HIV Insight (n = 10 873), Target Management Services (n = 2226) and Clinical Partners (n = 1505). Each database records the dates of starting and stopping individual antiretroviral agents over time, measurements of CD4 T-cell counts and HIV-RNA levels at approximately 6-monthly intervals, and the demographic characteristics of patients. The number, frequency and duration of different antiretroviral combinations over time and their relationship to stage of HIV-disease and demographic characteristics were explored. RESULTS: Over 2000 different combinations of antiretroviral agents are recorded. From 1987 onwards, the use of zidovudine increased, with 23% of patients receiving monotherapy by 1990. The majority of treated patients remained on monotherapy until the introduction of highly active antiretroviral therapy (HAART) in 1996. By 1999, the standard of care was HAART, with 84% of patients beginning antiretroviral therapy with HAART. Those of African American race (odds ratio 0.59) and funded by Medicaid (odds ratio 0.72) were significantly less likely to begin antiretroviral therapy on HAART. Until 1995, there was a significant decrease in CD4 T-cell count when starting antiretroviral therapy. No significant trend was observed in either CD4 T-cell count or viral load after this time. Those starting on HAART therapies were significantly less likely to stop or switch regimens than those on nucleoside reverse transcriptase inhibitor (NRTI)-only therapies (P < 0.001). CONCLUSIONS: Complex patterns of antiretroviral treatment are observed in this large population. Changes over time mirror the introduction of the new antiretroviral agents. PMID- 12534957 TI - Assessing limiting factors to the acceptance of antiretroviral therapy in a large cohort of injecting drug users. AB - OBJECTIVE: A comprehensive questionnaire was designed to assess the knowledge and understanding of injecting drug users (IDUs) regarding their HIV disease, and to determine any factors that may increase the acceptance of antiretroviral therapy (ART) by this group. RESULTS: Twenty percent of the total IDU cohort attending the GUIDE (GenitoUrinary Medicine and Infectious Diseases) clinic participated in the study. Fifty-two percent had been homeless in the past 5 years and 84% are unemployed. Seventy-two percent of patients did not complete second level education and 10% were illiterate. Fifty-one percent had siblings or parents with a history of injection drug misuse, and 25% had at least one sibling also HIV positive. Forty-seven percent started using drugs before the age of 13 years, and the most common initial drug was heroin (44%). Ninety-five percent had attended for methadone maintenance therapy (MMT), with 39% currently attending for daily therapy. The majority of patients were unable to simply explain or interpret CD4 cell counts (54%) and 'viral loads' (65%). Fifty-seven percent of patients were receiving highly active antiretroviral therapy (HAART). There was a statistically significant association between patients receiving HAART and both attendance at a primary care physician for methadone maintenance therapy (P = 0.005), and weekly take-outs of methadone (P = 0.005). There was also an association between adherence to HAART and attendance at a methadone maintenance clinic (P = 0.04). CONCLUSIONS: This study highlights the chaotic lifestyle and complex social background of the IDU. Such factors were not, however, associated with acceptance of HAART. The primary factor associated with both the acceptance of and adherence to HAART was regular and stable attendance for methadone therapy. PMID- 12534958 TI - Drug resistance mutations and outcome of second-line treatment in patients with first-line protease inhibitor failure on nelfinavir-containing HAART. AB - OBJECTIVES: To determine resistance mutations emerging in HIV-1-infected patients experiencing their first protease inhibitor (PI)-failure on nelfinavir-containing highly active antiretroviral therapy (HAART), and to assess virological response to rescue regimens. METHODS: Plasma HIV-1 RNA from 24 patients failing nelfinavir containing HAART was sequenced. Failure was defined as two consecutive measurements of viral load > 400 HIV-1 RNA copies/mL. Patients with previous failure on other PIs were excluded. Data on response to second-line treatment was extracted from patient files. RESULTS: At failure primary protease mutations were found in 14 patients (58%). Ten patients had D30N (38%), five patients had L90M (19%), two patients had V82A/F (8%) and two patients had M46I/L (8%). Two patients had both D30N and L90M. Pronounced increases of secondary protease mutations were seen at codon 88 (Delta: 33%), codon 36 (Delta: 30%) and codon 71 (Delta: 17%). Of eight patients with N88D, seven also harboured D30N (P < 0.01). Polymorphisms at codon 63 were detected at baseline in all patients who developed primary resistance mutations at failure (P < 0.01). On rescue regimens, 78% achieved viral loads below limit of detection (BLD). The presence of primary protease mutations was not associated with a higher risk of failure on second line treatment. CONCLUSION: In patients failing nelfinavir-containing HAART, D30N was detected frequently and L90M occasionally. A pronounced accumulation of the secondary protease mutations N88D, M36I, and A71V/T was found, and D30N was strongly associated with N88D. A high proportion of patients became undetectable on second-line treatment and the presence of primary resistance mutations did not negatively affect the outcome of rescue regimens. PMID- 12534959 TI - The use of calcium carbonate in nelfinavir-associated diarrhoea in HIV-1-infected patients. AB - OBJECTIVES: To evaluate the efficacy of oral calcium supplements in HIV-infected patients with nelfinavir (NFV)-associated diarrhoea, and to investigate the influence on the pharmacokinetics of nelfinavir and the active metabolite M8. METHODS: An open-label prospective trial with enrolment of 15 patients with NFV associated diarrhoea. Study subjects received either calcium carbonate or calcium gluconate/calcium carbonate in addition to highly active antiretroviral therapy (HAART), and were randomized to (i) calcium supplements for 14 days followed by 14 without calcium supplements, or (ii) 14 days without calcium supplements followed by calcium supplements for 14 days. Clinical endpoint was the severity of diarrhoea, graded and summarized for the specific 14 day-period. In the pharmacokinetic evaluation concentrations of NFV and M8 were measured before morning dosing, and 3 h after dosing. RESULTS: Nine patients were treated with calcium carbonate, and six with calcium gluconate/calcium carbonate. In the paired analysis, neither of the groups had a significant improvement in diarrhoea score when treated with calcium supplements (P = 0.34 and 0.46, respectively). We found no significant differences in the concentrations of NFV and M8 between the calcium and control periods. CONCLUSIONS: Oral calcium supplements did not significantly improve nelfinavir-associated diarrhoea. In the pharmacokinetic analysis calcium supplements did not induce major alterations in plasma concentrations of NFV and M8. PMID- 12534960 TI - Virological and immunological profiles among patients with undetectable viral load followed prospectively for 24 months. AB - OBJECTIVE: To quantify HIV-RNA in plasma, in lymphoid tissue and proviral DNA in peripheral blood mononuclear cells and to relate these to immunological markers among patients with plasma viral load counts of /= 1 measurement with 21 200 and 25% had >/= 1 sample with plasma HIV-RNA > 200 copies/mL. Lymphoid tissue viral load was low at enrolment and declined further during follow-up. Baseline HIV-DNA and immunoglobulin (IgA) differed significantly between the plasma viral load rebound groups (P < 0.05). CONCLUSION: In this cohort, selected solely on the basis of having a plasma viral load of or=50% at the end of the treatment phase) and partial-responders (seizure reduction of >or=50% at the end of the first month of treatment but 50% (OR 1.91), pump time (OR 1.67 per hour), open chamber (OR 1.95); and successfully eliminated the dummy variable gender (P = 0.6). This analysis indicates that the design of future stroke risk factor studies in the setting of cardiac surgery can and should adopt a neuroimaging evidence-based investigative approach of consolidating global with focal deficits. PMID- 12534998 TI - Cerebral vasomotor reactivity of the posterior circulation in patients with carotid occlusive disease. AB - We evaluated the hemodynamic features of the posterior circulation in patients with severe carotid stenosis by assessing and comparing cerebral vasomotor reactivity (VMR) in the middle cerebral (MCA) and vertebral arteries (VA) by transcranial Doppler and the Diamox (1 g acetazolamide i.v.) test. Sixty symptomatic and 111 asymptomatic patients with unilateral severe (>70%) internal carotid artery stenosis were studied. The VMR was 19.2 +/- 18.9% for the MCA ipsilateral to the stenosis and 27.3 +/- 17.4% on the contralateral side (P < 0.0001) for all patients. It was 18.2 +/- 23.2% for the VA ipsilateral to the stenosis and 19.7 +/- 21% on the contralateral side (P = NS). The symptomatic patients' VMR of the MCA on the side of stenosis and the opposite side were 19.2 +/- 17.6 and 29 +/- 17.2%, respectively (P < 0.03). The VMR of the VA remained similar (15.1 +/- 21 and 21.6 +/- 6%, respectively, P = NS). The asymptomatic patients' VMR of the MCA on the side of the stenosis was also lower (19.2 +/- 19.7 vs. 26.5 +/- 17.5% on the opposite side, P < 0.001). In contrast, the VMR in the VA was similar (19.8 +/- 21.4 and 18.7 +/- 19.5%, respectively, P < 0.6, NS). Thus, the VMR of the posterior circulation remained similar regardless of carotid stenosis and a symptomatic/asymptomatic course of carotid occlusive disease, suggesting an independent cerebral vascular reserve capacity of the posterior circulation. PMID- 12534999 TI - Olanzapine efficacy in the treatment of essential tremor. AB - We investigated the anti-tremor effect of olanzapine, a novel atypical antidopaminergic drug, in 37 patients with essential tremor (ET) in an open-label and prospective study by clinical scoring and patient self-evaluation. Olanzapine monotherapy appears to be efficacious for the treatment of ET. Further clinical trials with control groups are indicated to establish the efficacy of olanzapine in patients with ET. PMID- 12535000 TI - Subacute rhombencephalitis optica responsive to intravenous immunoglobulins. AB - A 53-year-old man presented with optic neuritis, followed within 10 weeks by a subacute progression of weakness, ataxia, and multiple cranial nerve palsies. Cranial magnetic resonance imaging demonstrated multiple T2-hyperintense lesions extending from the ponto-medullary junction to the thalamus and internal capsule. Cerebrospinal fluid analysis showed lymphocytic pleocytosis with elevated protein content. A brain biopsy revealed inflammatory changes. After a month long period of inexorable worsening, the patient was treated with intravenous immunoglobulins (2 g/kg). He responded to treatment, and recovered over the subsequent 3 months. The patient's response to treatment suggests that intravenous immunoglobulins should be considered in patients with inflammatory brain disorders. PMID- 12535001 TI - EMG myokymia as a cause of ptosis in hypothyroidism. AB - Ptosis is known to be associated with thyroid disorders. We describe two biochemically corrected hypothyroid patients presenting with isolated bilateral ptosis. EMG of the orbicularis oculi showed continuous grouped motor unit potentials. In the absence of obvious aetiology, it is hypothesised that focal demyelination of terminal branches to the orbicularis oculi may play a role in the generation of the discharges. PMID- 12535002 TI - Effect of vagal nerve stimulation on patients with bitemporal epilepsy. AB - Patients with bitemporal epilepsy are characterized by the existence of independent bitemporal seizure onset zones. The aim of this study was to evaluate the effect of chronic vagal nerve stimulation (VNS) on eight patients with bitemporal epilepsy. We demonstrated the gradually increased effect of VNS on the reduction of seizures as compared with baseline seizure frequency in patients with bitemporal epilepsy. The average seizure reduction increased from 4.2% at the 3-month follow-up visit to 18.2, 34.4 and 42.2% at the 6, 12 and 18-month follow-up visits. Similarly, a >or=50% reduction of complex partial seizures was reported at the 3-month follow-up visit in no patients (0%); at the 6-month follow-up visit in one patient (12.5%); at the 12-month follow-up visit in three patients (37.5%); and at the 18-month follow-up visit in five patients (62.5%). These data demonstrate the positive and long-lasting effect of VNS on seizure reduction in patients with intractable bitemporal epilepsy. The main mechanism of this chronic effect is not fully understood. PMID- 12535003 TI - 'Gender gap' in multiple sclerosis: magnetic resonance imaging evidence. AB - The authors evaluated the gender difference in the magnetic resonance imaging characteristics of the lesions occurring in the brain of 413 multiple sclerosis (MS) patients. Men had fewer contrast-enhancing lesions (P = 0.01), but a higher proportion of lesions evolving into 'black holes' (P = 0.001), when compared with women. Thus, our data indicate that men with MS are prone to develop less inflammatory, but more destructive lesions than women. This study results provides support for a modulation of the MS pathological changes by gender. PMID- 12535004 TI - Intracerebral haemorrhage as a manifestation of Lyme neuroborreliosis? AB - Lyme Neuroborreliosis (LNB) has repeatedly been reported to cause cerebral vasculitis. However, there is no reliable information about the incidence of cerebral vessel affection. The majority of reports deal with ischaemic consequences, and there are a few reports of subarachnoid haemorrhage (SAH). We report a case of otherwise unexplained intracerebral haemorrhage (ICH) where clinical and laboratory tests have shown LNB. Late stage LNB might not only cause ischaemic, but haemorrhagic stroke as well. PMID- 12535006 TI - Camptocormia as a form of dystonia in Parkinson's disease. PMID- 12535005 TI - Yes/no reversals as neurobehavioral sequela: a disorder of language, praxis, or inhibitory control? AB - This study identifies a linguistic phenomenon suggestive of damage to fronto subcortical circuitry. Our objective was to determine the occurrence and neuroradiological/neurobehavioral correlates of yes/no reversals in corticobasal degeneration (CBD), and document occurrence of reversals in other neurological conditions. In a prospective study, we evaluated 34 CBD patients using a neuropsychologic battery and magnetic resonance imaging. Patients were subdivided into two groups: those with (n = 11) and without (n = 23) yes/no reversals. In a retrospective study conducted during the period of 1991-2001, we identified 33 patients for whom yes/no reversals occurred to compare correlates with prospective study findings. In the prospective study, 11 patients (32.3%) had yes/no reversals. Significant between-group differences were found in scores of lexical fluency (P = 0.02) and prehension (P = 0.03). Prehension scores correlated with facial praxis (P < 0.0001) and upper limb praxis scores (P < 0.0001) in the yes/no reversal group only. In the retrospective study, nine CBD patients and 24 non-CBD patients had yes/no reversals, with damage to fronto subcortical areas present in all patients. Results suggest an association with deficits in mental flexibility and inhibitory control. High within-group correlations of lexical fluency and prehension with praxis scores suggest a relationship of yes/no reversals with multiple factors. PMID- 12535007 TI - How specific are the pontine MRI hyperintensities (the cross sign)? PMID- 12535008 TI - Acute cervical hyperextension-hyperflexion injury may precipitate and/or exacerbate symptomatic multiple sclerosis. PMID- 12535022 TI - Ultraviolet radiation enhances both the nodular and ulcerative forms of Mycobacterium ulcerans infection in a Crl:IAF(HA)-hrBR hairless guinea pig model of Buruli ulcer disease. AB - BACKGROUND: Ultraviolet radiation (UV) pre-exposure enhances intracellular mycobacterial infections, however, its effect upon the pathogenesis of the extracellular Mycobacterium ulcerans parasite had not been previously examined. The hypothesis tested was that UV pre-exposure enhances both the nodular and ulcerative forms of M. ulcerans infection in the Crl:IAF(HA)-hrBR hairless guinea pig. METHODS: Groups of five animals were exposed to total cumulative UV doses of 0 (control), 3 or 30 kJ/m2 followed 3 days later by subcutaneous infection with 3 x 10(4) CFU of M. ulcerans in order to induce the nodular form of the disease. The resultant nodules were then measured for the next 22 days. The experiment was then repeated using intradermal infection with 2 x 10(6) CFU in order to induce the ulcerative form of the disease. The resultant ulcers were measured for the next 30 days. In both experiments, the animals were tested for delayed-type hypersensitivity (DTH) reactivity to Burulin-S as a marker of the onset of the reactive phase of the disease. RESULTS: Following low inoculum subcutaneous infection, distinct, well-demarcated, subcutaneously situated skin nodules were present at infected skin sites between 7 and 22 days post-infection. Between days 14 and 21, the mean nodule diameters of the UV irradiated groups were significantly (P < 0.03) greater than that of the control group. UV pre-exposure resulted in significant (P < 0.035) suppression of DTH responses to Burulin-S challenge. High inoculum intradermal infection resulted in the development of ulcerative lesions. Between 10 and 30 days post-infection, the mean lesion diameters and mean ulcer development times of UV irradiated groups were significantly (P < 0.05) greater than those of the controls. However, UV irradiation did not affect DTH responses to Burulins in the high inoculum experiment. In both experiments, the lesions were histologically consistent with human Buruli ulcer disease. These results demonstrate that UV pre-exposure results in enhanced M. ulcerans infection in the hairless guinea pig model of Buruli ulcer disease and suggest that UV exposure may be a relevant factor in the pathogenesis of human forms of the disease. PMID- 12535023 TI - The effect of amniotic membrane extract on the expression of iNOS mRNA and generation of NO in HaCaT cell by ultraviolet B irradiation. AB - BACKGROUND/PURPOSE: Amniotic membrane (AM) is the innermost fetal membrane, which contains several proteinase inhibitors and expresses several growth factors. Nitric oxide (NO) has been implicated in the pathogenesis of various inflammatory diseases including sunburn and ultraviolet induced erythema. The expression of inducible nitric oxide synthase (iNOS) is up regulated by UVB irradiation and inhibited by TGF-beta and EGF-beta. We evaluated the effect of AM extract on the expression of iNOS mRNA by UV irradiation in HaCaT cell (immortalized human keratinocyte cell line). METHODS: HaCaT cells were irradiated UVB 30 mJ/cm2 and AM extract was added. The iNOS mRNA was isolated by RT-PCR and NO production was assessed by spectrophotometric method based on Griess reaction. RESULTS: The expression of iNOS mRNA was induced by UVB irradiation in HaCaT cell and the expression of iNOS mRNA was higher at 48 h than that at 24 h. AM extract down regulated the induction of iNOS mRNA in HaCaT cell by UVB irradiation. NO generation was increased by UVB irradiation, but down regulated by AM extract treatment in HaCaT cells. CONCLUSION: These results assured that the expression of iNOS mRNA and generation of NO are up regulated by UVB irradiation and showed that AM extract down regulated the induction of iNOS mRNA and decreased generation of NO by UVB irradiation. PMID- 12535024 TI - The mutagenic effect of ultraviolet-A1 on human skin demonstrated by sequencing the p53 gene in single keratinocytes. AB - BACKGROUND: Sun exposure is accepted as the major risk factor for developing skin cancer, the most common cancer in the western world. Ultraviolet-B (UV-B) radiation is considered the causative agent, but recently several findings suggest a role also for ultraviolet-A (UV-A) radiation. Repeated suberythemal doses of ultraviolet-A1 (UV-A1) on healthy human skin induce an increase of p53 immunoreactive cells in epidermis, which may indicate cell cycle arrest and/or occurrence of p53 mutations. METHODS: We have investigated the possible mutagenic effect of UV-A1 on skin by sequencing exons 4-11 and adjacent intron sequence of the p53 gene in immunoreactive single cells from three healthy individuals. Previously unexposed buttock skin was irradiated three times a week for 2 weeks with physiological fluences (40 J/cm2) of UV-A1. Punch biopsies were taken before and at different time-points after the exposure, and from these single p53 immunoreactive cells were isolated by using laser-assisted microdissection. RESULTS: Three mutations--all being indicative of oxidative damage and most likely related to UV-A exposure--were found among the 37 single cells from exposed skin, whereas no mutations were found in the 22 single cells taken before exposure. CONCLUSIONS: The findings indicate a mutagenic effect of low-dose UV-A1 on healthy human skin, which further demonstrates the importance of considering UV-A when taking protective measures against skin cancer. PMID- 12535025 TI - Skin cancer and ultraviolet-B radiation under the Antarctic ozone hole: southern Chile, 1987-2000. AB - BACKGROUND: Punta Arenas, Chile, the southernmost city in the world (53 degrees S), with a population of 154,000, is located near the Antarctic ozone hole (AOH) and has been regularly affected by high levels of ultraviolet-B (UV-B) radiation each spring for the last 20 years. Large increases in UV-B associated with the AOH have been measured with increases in UV-B at 297 nm of up to 38 times those of similar days with normal ozone. Recently we reported significant increases in sunburns during the spring of 1999 on days with low ozone because of the AOH. METHODS: A surveillance of skin cancers occurring from 1987 to 2000 was performed. Age, sex, location, type of skin cancer and skin phototype were recorded. A Brewer Spectrophotometer was used in order to obtain in situ measurements of ozone and UV-B. Total Ozone Mapping Spectrometer (TOMS) data from National Aeronautics and Space Administration (NASA) was used in order to establish pre-ozone hole climatology. RESULTS: Ozone levels as low as 145 DU (Dobson Units) were recorded, a 56% decrease in ozone, and UV-B levels up to 4.947 J/m2. These levels are close to summertime levels at mid latitudes. For the 14-year period--from 1987 to 2000--173 cases of skin cancer were diagnosed, 65 during the first 7 years, 108 during the second, an increase of 66%. Cutaneous malignant melanoma (CMM), 19% of the cases, increased by 56%, raising the rate from 1.22 to 1.91 per 100,000. Non-melanoma skin cancer (NMSC), 81% of the total, increased the rate from 5.43 to 7.94 per 100 000 (P < 0.05), a 46% increase. Patients with CMM and NMSC had skin phototypes I-II in 59% and 54% of cases, respectively. Days with more than 25% ozone loss occurred in 143 days during the last 20 springs. Significant increases of UV-B were observed under ozone hole conditions, especially around 300 nm, the most carcinogenic wavelengths. CONCLUSIONS: Highly unusual ozone loss and UV-B increases have occurred in the Punta Arenas area over the past two decades resulting in the non-photoadapted population being repeatedly exposed to an altered solar UV spectrum with a greater effectiveness for erythema and photocarcinogenesis. This phenomenon has not previously been reported over other populated areas and an additional increase in the skin cancer rate attributable to the AOH may be occurring. Research on the clinical and subclinical impact of these abnormalities is urgently needed. PMID- 12535026 TI - Polymorphous light eruption in African Americans: pinpoint papular variant. AB - BACKGROUND: Polymorphous light eruption (PMLE) is the most common chronic idiopathic photodermatosis usually manifesting as a papular eruption (3-6 mm), with several other morphological variants described. METHODS: Between June 1998 and August 2001, nine patients presented with complaints of a pruritic pinpoint papular eruption associated with sun exposure. A detailed history and complete skin examination were performed along with a skin biopsy if active lesions were present. Phototesting to ultraviolet-A (UV-A), ultraviolet-B (UV-B) and visible light was performed in four patients. Antinuclear antibody (ANA) testing was performed in three patients. The diagnosis of PMLE was made based on the history, morphology of the lesions, results of phototesting and skin biopsy if available. RESULTS: In all patients, pinpoint papules (1-2 mm) were observed on sun-exposed areas, sparring the face and flexural surfaces. All patients were African American women with skin type IV-VI and a mean age of 39.3 years (range 21-52 years). Phototest results were normal in three patients; one patient, who was on glyburide, had a decreased minimal erythema dose to UV-A. ANA testing was negative. Two histopathologic patterns were observed: (i) focal lichenoid and perivascular lymphohistiocytic infiltrate with red blood cell extravasation in four specimens and (ii) superficial and deep interstitial lymphocytic infiltrate with papillary dermal edema in the remaining three specimens. All patients responded to topical corticosteroids, broad-spectrum sunscreens and antihistamines. CONCLUSION: Recognition of this pinpoint papular variant of PMLE in dark-skinned individuals is important in the evaluation and management of these patients. PMID- 12535027 TI - Long-term treatment of erythropoietic protoporphyria with cysteine. AB - A Phase III 3 year placebo-controlled trial, in which patients were blinded as to when placebo was given, was conducted to determine if the administration of L cysteine-HCl was effective in preventing or ameliorating the photosensitivity associated with erythropoietic protoporphyria. Forty-seven patients participated in the trial. Placebo was administered in the first month of the trial and then 500 mg of cysteine was given in two divided doses per day for the duration of the study. To measure efficacy, baseline and follow-up visit history forms and light exposure diaries were filled in by the patients, and those seen in Boston were phototested. Analysis of the data from the questionnaires, the minutes of symptom free light exposure recorded in diaries and history forms and the results of the phototests showed that cysteine increased the time of symptom-free light exposure to a statistically significant degree. PMID- 12535029 TI - Chronic actinic dermatitis secondary to simvastatin. PMID- 12535030 TI - A provocative case: phototesting does not reproduce the lesions of actinic granuloma. PMID- 12535031 TI - Do we need a new muscle relaxant in paediatrics? PMID- 12535032 TI - Tracheomalacia and bronchomalacia in children: pathophysiology, assessment, treatment and anaesthesia management. AB - Tracheomalacia and bronchomalacia are becoming increasingly well recognized. Although pathologically benign conditions, they are responsible for considerable morbidity, occasional mortality and significant difficulties in the operating theatre and intensive care unit. We performed an extensive literature search to identify causal associations, methods of clinical and investigative assessment, treatment modalities and anaesthetic experience with these conditions. PMID- 12535033 TI - Ventilatory effects of morphine infusions in cyanotic versus acyanotic infants after thoracotomy. AB - BACKGROUND: Previous studies show that the age of an infant affects morphine clearance but that ventilatory effects from morphine infusions are similar at the same morphine steady-state concentration in infants and children after surgery. The presence of cyanotic heart disease in infants receiving postoperative morphine infusions was studied for its potential effect on ventilatory effort. METHODS: Twenty infants under 90 days of age (10 cyanotic, 10 acyanotic) received morphine intravenous infusions following thoracotomy. PaCO2 during spontaneous breathing, CO2 rebreathing trials during morphine infusions and time to wean from assisted mechanical ventilation after decreasing infusion rates were compared. Analgesia scoring in the first postoperative 24 h was also compared between cyanotic and acyanotic infants. RESULTS: PaCO2 during spontaneous breathing and CO2 response curve slopes were not different between groups. The time to wean from assisted mechanical ventilation after decreasing morphine infusion rates was not statistically different (median 6.9 h versus 5.8 h in cyanotic and acyanotic, respectively). Median morphine concentrations were higher in cyanotic infants at the time of ventilatory testing (32 versus 20.5 ng x ml-1 in cyanotic versus acyanotic infants, respectively, P = NS). Both groups had good pain relief. Steady-state morphine clearance rates showed two- to three-fold variability between infants. They were not statistically different in cyanotic versus acyanotic infants but tended to be lower in the cyanotic. Morphine clearance increased significantly with increasing age. CONCLUSIONS: Morphine i.v. infusions given to infants following thoracotomy show the same ventilatory effects in infants with cyanotic heart disease as in acyanotic infants. Age is the more important determinant of morphine response by its effect on morphine clearance than cyanosis in infants aged less than 3 months. PMID- 12535034 TI - Haemodynamic changes during low-pressure carbon dioxide pneumoperitoneum in young children. AB - BACKGROUND: Both mechanical and pharmacological effects may contribute to the haemodynamic consequences of carbon dioxide (CO2) pneumoperitoneum. The aim of the present study was to evaluate the haemodynamic effects of low-pressure pneumoperitoneum [intra-abdominal pressure (IAP) 5 mmHg] in young children (< 3 years). METHODS: Thirteen children, aged 6-36 months, ASA physical status I-III, who were scheduled for laparoscopic fundoplication for gastro-oesophageal reflux were investigated in the head-up position (10 degrees ). Noninvasive thoracic electrical bioimpedance cardiac index (CI), stroke volume index (SVI), heart rate (HR), mean arterial pressure (MAP) and peak inspiratory pressure (PIP) were recorded, together with PetCO2 and PaCO2 at five time points: before insufflation, 20, 35 and 70 min after start of CO2 insufflation and 12 min after desufflation. During insufflation, minute ventilation was not adjusted and the IAP was maintained at 5 mmHg. RESULTS: During insufflation, PetCO2 increased from 29 +/- 4 to 37 +/- 5 mmHg (P < 0.001) and PaCO2 increased from 31 +/- 4 to 39 +/- 5 mmHg (P < 0.01). CI increased from 2.39 +/- 0.86 to 2.92 +/- 0.94 l x min-1 x m2 (P < 0.01), HR increased from 108 +/- 10 to 126 +/- 22 b x min-1 (P < 0.01), MAP increased from 52 +/- 10 to 63 +/- 9 (P < 0.05) and PIP increased from 16 +/- 3 to 18 +/- 3 cm H2O (P < 0.001). There were no changes in SVI and arterial oxygen saturation. CONCLUSIONS: We conclude that low-pressure CO2 pneumoperitoneum (with IAPs not exceeding 5 mmHg) for laparoscopic fundoplication in infants and children does not decrease their cardiac index. PMID- 12535035 TI - Cerebrovascular carbon dioxide reactivity in children anaesthetized with propofol. AB - BACKGROUND: Propofol, by virtue of its favourable pharmacokinetic profile, is suitable for maintenance of anaesthesia by continuous infusion during neurosurgical procedures in adults. It is gaining popularity for use in paediatric patients. To determine the effects of propofol on carbon dioxide cerebrovascular reactivity in children, middle cerebral artery blood flow velocity was measured at different levels of endtidal (PECO2) by transcranial Doppler sonography. METHODS: Ten ASA I or II children, aged 1-6 years undergoing elective urological surgery were enrolled. Anaesthesia comprized propofol aimed at producing an estimated steady-state serum concentration of 3 microg x ml-1 and a caudal epidural block. PECO2 was adjusted randomly in an increasing or decreasing fashion between 3.3, 5.2 and 7.2 kPa (25, 40 and 55 mmHg) with an exogenous source of CO2 while maintaining ventilation parameters constant. RESULTS: Cerebral blood flow velocity increased as PECO2 increased from 3.3 to 5.2 kPa (25-40 mmHg) (P < 0.001) and from 5.2 to 7.2 kPa (40-55 mmHg) (P < 0.001). Mean heart rate and blood pressure did not change significantly. CONCLUSIONS: This study demonstrates that cerebrovascular CO2 reactivity is maintained over PECO2 values of 3.3, 5.2 and 7.2 kPa (25, 40 and 55 mmHg) in healthy children anaesthetized with propofol. PMID- 12535036 TI - The effectiveness and safety of spinal anaesthesia in the pyloromyotomy procedure. AB - BACKGROUND: Hypertrophic pyloric stenosis is a relatively common disorder of the gastrointestinal tract in infancy, causing projectile vomiting and metabolic abnormalities. Surgical management in the form of pyloromyotomy under general anaesthesia has been reported as safe for relieving the obstructed bowel. A number of studies have demonstrated the advantages of spinal anaesthesia over general anaesthesia in high risk infants undergoing minor infraumbilical surgery. The purpose of this study was to evaluate spinal anaesthesia as an alternative option to general anaesthesia in infants undergoing pyloromyotomy. METHODS: Twenty-five infants undergoing pyloromyotomy under spinal anaesthesia were studied. Haemodynamic and respiratory parameters were noted before performing the spinal block, 5 min after the spinal block, and every 10 min after performing the spinal block; for a total period of 30 min. The spinal block was performed using spinal isobaric bupivacaine 0.5%, 0.8 mg.kg-1. Blood pressure, heart rate, respiratory rate and oxygen saturation values were recorded. RESULTS: The sensory levels achieved ranged between T3-T5 thoracic segments within 6-8 min and were sufficient to perform surgery in 23 cases. There were no statistically significant differences in the oxygen saturation, systolic blood pressure and respiratory rate compared with before the spinal block and after 5, 10, 20 and 30 min. CONCLUSIONS: This study proposes that spinal anaesthesia is an alternative option to general anaesthesia in infants undergoing pyloromyotomy, and should be considered in infants undergoing pyloromyotomy. PMID- 12535037 TI - A comparison between local anaesthetic dorsal nerve block and caudal bupivacaine with ketamine for paediatric circumcision. AB - BACKGROUND: Ketamine has been shown to prolong analgesia produced by caudal local anaesthetic block and is now in common use. This study compares caudal block using bupivacaine/ketamine with dorsal nerve block of the penis. METHODS: Sixty boys undergoing elective circumcision were given either 0.5 ml x kg-1 of bupivacaine 0.15% with ketamine 0.5 mg x kg-1 (n = 30) or dorsal nerve block of the penis with bupivacaine 0.5% (n = 30) as a supplement to general anaesthesia. Postoperative pain was assessed by parents using a modified objective pain score, and the time taken to first requirement of analgesia was recorded. Motor weakness, time to first micturition, postoperative nausea and vomiting (PONV), eating habits, sleep disturbance and behaviour were also assessed. RESULTS: There was no difference between the groups in time to first requirement for analgesia or number of doses of paracetamol given in the first 24 h. Almost half the boys in the caudal group had motor weakness, and there was a significant increase in time to first micturition in that group. There was no difference between the groups in PONV, eating, sleeping or behavioural disturbance. CONCLUSIONS: Caudal anaesthesia with bupivacaine/ketamine does not confer any advantage over a dorsal nerve block with the doses used in this study. Because of the higher incidence of side-effects and technique failure in the caudal group, dorsal nerve block is perhaps the preferred technique. PMID- 12535038 TI - The use of magnesium to prevent laryngospasm after tonsillectomy and adenoidectomy: a preliminary study. AB - BACKGROUND: Laryngospasm is the most common cause of upper airway obstruction after tracheal extubation. Magnesium has a central nervous system depressant property, which contributes to the depth of anaesthesia. It also has calcium antagonist properties, which provide muscle relaxation. In this study, we aimed to determine the effect of magnesium on preventing laryngospasm. METHODS: After approval of the Ethics Committee and informed parental consent, 40 patients, ASA I-II, aged 3-12 years, who were scheduled for tonsillectomy or/and adenoidectomy, were randomly divided into two groups. Anaesthesia was induced with sevoflurane, lidocaine 1 mg x kg-1, alfentanil 10 micro g x kg-1, vecuronium 0.1 mg x kg-1 and maintained with sevoflurane 2% and 60% nitrous oxide in oxygen. After intubation, patients in group I received 15 mg.kg-1 magnesium in 30 ml 0.9% NaCl over 20 min. Patients in group II received 0.9% NaCl alone in the same volume. After reversal of neuromuscular blockade, all patients were extubated at a very deep plane of anaesthesia. The incidence of laryngospasm was determined until the time of discharge from the postanaesthesia care unit. RESULTS: Although laryngospasm was not observed in group I, it was observed in five patients in group II (25%). The incidence of laryngospasm in group II was significantly higher than group I. The plasma magnesium concentrations were significantly higher in group I than group II. CONCLUSIONS: We found a significant decrease in the incidence of laryngospasm in paediatric patients receiving magnesium. It is suggested that the use of intravenous magnesium intraoperatively may prevent laryngospasm. PMID- 12535039 TI - Haemodynamic changes during high spinal anaesthesia in children having open heart surgery. AB - BACKGROUND: This prospective series examined the haemodynamic effects of high spinal anaesthesia in combination with light general anaesthesia in infants and children undergoing open heart surgery who were candidates for immediate or early postoperative extubation. METHODS: After midazolam premedication and sevoflurane inhalation induction, 30 patients, aged 7 months to 13 years, who were undergoing open heart surgery, received spinal anaesthetics with 0.5% tetracaine D10 mixed with morphine. The spinal blocks were placed at the L2,3 or L3,4 interspace with cephalad spread being promoted by positioning the patient in 30 degrees of Trendelenburg for a minimum of 10 min. Maintenance of anaesthesia was with isoflurane 0.2-0.5% in 70% nitrous oxide to maintain heart rate and blood pressure within 20% of postinduction baseline values. Haemodynamic values were recorded at predetermined timed intervals and intraoperative events up to and including aortic cannulation. For analysis of the data, patients were divided into four age groups (< 1 years, 1-3 years, 4-6 years and > 7 years). RESULTS: Haemodynamic stability was demonstrated in all four age groups. Statistically significant slowing of the heart rate did occur in the groups older than 1 year at 25 min, although clinically significant bradycardia requiring treatment never occurred. Hypotension did occur during specific surgical manipulations but recovered spontaneously. Atropine, fluid boluses and vasopressors were never used. At the conclusion of surgery, all patients met extubation criteria and could move all four extremities. CONCLUSIONS: High spinal anaesthesia with hyperbaric tetracaine and morphine in combination with light general anaesthesia is well tolerated haemodynamically by the paediatric population studied. PMID- 12535040 TI - Propofol anaesthesia and metabolic acidosis in children. AB - BACKGROUND: We aimed to investigate the effect of propofol infusion anaesthesia on acid-base status and liver and myocardial enzyme levels of children during short-term anaesthesia. METHODS: Thirty-six children, aged 3-12 years, were randomized into two groups. In group P (n = 18), induction and maintenance were performed with propofol, 3 mg x kg-1 and 20, 15 and 10 mg x kg-1 x h-1, respectively. In group H (n = 18) following induction with 5 mg x kg-1 thiopenthal, anaesthesia was maintained with 2-3% halothane. Blood samples were obtained following anaesthesia induction and 30, 60 and 120 min after discontinuation of anaesthesia. RESULTS: There was no difference in lactate dehydrogenase, myocardial creatininephosphokinase, aspartate aminotransferase, alanine aminotransferase and cholesterol levels between and within the groups. All postoperative triglyceride levels were higher and pH levels were lower in group P than group H (P < 0.05) and there was no difference within the groups. CONCLUSIONS: In these healthy patients, short-term use of propofol did not result in significant acidaemia, nor alterations in hepatic or myocardial enzyme levels. PMID- 12535041 TI - Combining low inspired oxygen and carbon dioxide during mechanical ventilation for the Norwood procedure. AB - BACKGROUND: Staged reconstruction has become the preferred approach for treating neonates with hypoplastic left heart syndrome (HLHS). The haemodynamic instability of a single ventricle providing blood flow in parallel to the systemic and the pulmonary circulation, combined with the effects of cardiopulmonary bypass (CPB), results in precarious perioperative conditions. The two ventilatory manoeuvres commonly used for increasing pulmonary vascular resistance are (i) decreasing the fraction of inspired oxygen to < 0.21 and (ii) adding carbon dioxide (CO2) to the ventilatory circuit. Whether molecular nitrogen (N2) or CO2 is used in these situations is a matter of physician and institutional preference. The effect of the two modalities in combination has not been studied in depth. METHODS: This prospective observational study was designed to look at the effects of adding inhaled CO2 to children with HLHS who were already on a hypoxic mixture during the immediate perioperative period. RESULTS: Twelve suitable neonates were enrolled in the study. Combining the two ventilatory modalities had an additive effect. The effect was more significant in the prebypass (83% of patients) compared with the postbypass period (25% of patients). CONCLUSIONS: Low inspired oxygen and CO2 have an additive vasoconstrictive effect on the pulmonary vessels. The use of both of these ventilatory manoeuveres is less effective postoperatively compared with the prebypass period. PMID- 12535042 TI - Rapid emergence does not explain agitation following sevoflurane anaesthesia in infants and children: a comparison with propofol. AB - BACKGROUND: Emergence agitation in children is frequently associated with the use of the new highly insoluble volatile anaesthetics. Rapid emergence has been cited as one of the possible causes. Propofol also permits rapid emergence from general anaesthesia but is not associated with agitation. METHODS: The emergence characteristics of children receiving sevoflurane and propofol anaesthesia were examined. After induction with sevoflurane, 53 children, aged 2-36 months, who were undergoing ambulatory surgery, were randomized to receive maintenance anaesthesia with either sevoflurane or propofol. Introperative analgesia with either 2 micro g x kg-1 of intravenous fentanyl or a caudal block with 0.25% bupivacaine was supplied according to surgical procedure. An observer blinded to anaesthetic technique recorded the time to achieve extubation and recovery and assessed emergence behaviour. Data were analysed using Wilcoxon scores, Kruskal Wallis test, chi-square and multiple regression analysis. RESULTS: The results showed that the time to extubation and recovery were similar between the two study groups, but that emergence agitation was significantly higher in the sevoflurane group compared with the propofol group. No relationship between analgesic technique and agitation scores was found. CONCLUSIONS: Although both sevoflurane and propofol allow for rapid emergence from general anaesthesia, only sevoflurane is associated with a high incidence of emergence agitation in infants and young children. Rapid emergence does not fully explain this phenomena. PMID- 12535043 TI - Congenital laryngotracheo-oesophageal cleft: successful ventilation with the Laryngeal Mask Airway. AB - A congenital laryngotracheo-oesophageal cleft is a rare airway malformation which results from incomplete separation of the larynx and trachea from the hypopharynx and oesophagus. Patients usually present with stridor, aspiration and cyanosis associated with feeding. For early diagnosis, a high index of suspicion is needed. Unless an appropriate diagnostic approach is taken, the diagnosis can be missed. The successful ventilation of a neonate with the Laryngeal Mask Airway is described. PMID- 12535044 TI - Perioperative care of the child with the Johanson-Blizzard syndrome. AB - The Johanson-Blizzard Syndrome (JBS) is an autosomal recessive disorder with a characteristic phenotype, including dwarfism, a beaked nose with aplastic alae nasi, a high forehead, mid-line ectodermal scalp defects with sparse hair and absent eyelashes/eyebrows, prominent scalp veins, low set ears, a large anterior fontanelle, micrognathia, thin lips, absent permanent dentition and microcephaly. In addition to the characteristic facial features, associated conditions include congenital heart disease, exocrine/endocrine pancreatic dysfunction, hypothyroidism, hypopituitarism, mental retardation, sensorineural hearing loss and vesico-ureteral reflux. A case is presented and the potential anaesthetic implications of this syndrome are discussed. PMID- 12535045 TI - Case report of a 13-year-old struck by lightning. AB - Lightning strikes kill 1,000 people per year worldwide. Cardiac arrests resulting from lightning strikes have good survival rates but there is a significant degree of morbidity amongst the survivors. This is the case report of a 13-year-old boy who had a cardiac arrest following a direct lightning strike, and his subsequent management. PMID- 12535046 TI - Popliteal pterygium syndrome with syngnathia. AB - We report a case of perioperative management of a neonate with popliteal pterygium syndrome complicated by interalveolar syngnathia. Syngnathia were excised in the operating room without a major anaesthetic. We discuss our management of this case, as well as other possible strategies to secure the airway in neonates with syngnathia. We also reviewed the literature regarding airway management in presence of syngnathia in similar situations. PMID- 12535047 TI - Anaesthetic considerations for a 2-month-old infant with suspected complex I respiratory chain deficiency. PMID- 12535048 TI - Anaesthetic/analgesic management of extracorporeal shock wave lithotripsy in paediatric patients. PMID- 12535049 TI - Airway management in a baby with femoral hypoplasia--unusual facies syndrome. PMID- 12535050 TI - A clinical procedure to predict the value of temporary occlusion therapy in keratoconjunctivitis sicca. AB - PURPOSE: To evaluate the effects of dissolvable collagen punctal plugs on the symptoms, tear stability and volume in aqueous deficient dry eyes. METHODS: Sixty two aqueous deficient dry eye patients of mixed aetiology underwent lacrimal punctal occlusion with dissolvable collagen plugs. The subjects were randomly allocated to one of two treatment groups: group I (n = 36) had their lower puncta occluded and group II (n = 26) had both their upper and lower puncta occluded. The effectiveness of this treatment was clinically assessed by (1). scoring subject symptoms and (2). measuring the tear parameters of tear thinning time (TTT) and tear meniscus height (TMH) as indicators of tear stability and volume, respectively. Following baseline measurements, patients were reviewed at time intervals of 5 and 12 days post-occlusion. A group of age- and gender-matched normals (n = 45) was recruited for comparison (group III). RESULTS: Tear volume and stability were significantly higher in group III compared with I and II at baseline. In the treated groups on both days 5 and 12: (1). symptom score reduced significantly from a median value of 7 to 3 (p = <0.001); (2). tear stability increased significantly from a median value of 3 to 5 s by day 5 (p or=0.2 were between 5 and 25% of evaluations. Caution should be exercised when using stereoscopic and non-stereo digital evaluations of CD ratio interchangeably to assess longitudinal progression in a multi-clinician setting. PMID- 12535058 TI - Cardinal points and image-object magnification with an accommodative lens implant (1 CU). AB - A simple mathematical method for the determination of the cardinal points of pseudophakic eyes after implantation of an accommodative intraocular lens [posterior chamber intraocular lenses (PCIOL)] is presented. The purpose of this study was to explore the changes during pseudophakic accommodation (PAC) in (1). the positions of the cardinal points, (2). the distance of the object conjugate with the retina, and (3). the image-object magnification. These theoretical accommodation data are compared with clinical measurements. METHODS AND PATIENTS: Using biometrical measurements of the axial length, equivalent power of the cornea and the anterior chamber depth (ACD) in the non-accommodated state we used linear geometric optics for determination of the cardinal points and object distance as well as lateral magnification (the ratio of image to object size). With the measurement of ACD decrease (following pharmacological stimulation of the ciliary muscle with 2% pilocarpine eye drops) we determined the changes of the cardinal points and magnification to assess PAC amplitude from the shortening of the object distance. Calculated values of PAC amplitude were compared with the respective measured values derived from amplitude measures by accommodometer, defocusing and streak retinoscopy. We analysed the results of a prospective study on 35 eyes of 28 patients after cataract surgery (target refraction: -0.2 D) and accommodative PCIOL implantation (1 CU, Human Optics AG, Erlangen, Germany) 3 months after surgery. RESULTS: After pilocarpine eye drops, ACD (mean +/- S.D., range; median) decreased by 0.88 +/- 0.48 mm (0.51-1.91; 0.66). Distance of the in-focus object decreased from the non-accommodated state (-5.62 +/- 1.83 m, -25 to -1.1; -4.83 m) to the accommodated state (ACD decrease) (-0.81 +/- 0.21, -2.11 to -0.65; -0.79 m). For a theoretical ACD decrease of 1.0 mm (the intrinsic limitation of the PCIOL design) it was -0.59 +/- 0.28, -1.31 to -0.51; -0.63 m and resulted in an objective accommodative response of 1.49 +/- 0.16, 1.21-1.81; 1.46 D, depending on the actual geometry of the individual eye. On average, magnification as induced by PAC in contrast to that induced by adequate spectacle addition differed by only about 1%. Accommodation measured with defocusing and the accommodometer correlated significantly with the theoretical value based on IOLMaster measurement of ACD decrease (r = 0.752, p = 0.005 and r = 0.676, p = 0.02). Likewise, accommodation measured with streak retinoscopy correlated weakly with the theoretical value based on IOLMaster ACD decrease (r = 0.465, p = 0.05). CONCLUSIONS: Using geometrical optics, PAC can be derived from the biometric data of the eye and the measured ACD decrease. This approach may be an additional indicator for the accommodative response in pseudophakic patients and may allow a subdivision of the measured accommodation into true PAC and pseudoaccommodation, for example, because of increased depth of focus induced by pupillary constriction. PMID- 12535059 TI - The effect of changing from glasses to soft contact lenses on myopia progression in adolescents. AB - At the end of a clinical trial of bifocals as myopia treatment, subjects were allowed to select any type of optical correction they wished and were asked to return in 1 year. This report gives results of that last examination with emphasis on how progression rates differed between those remaining in their original type of glasses compared with those who switched to soft contact lenses. We found that myopia progressed at an age-adjusted average rate of 0.74 D in 19 children who switched to soft contact lens wear compared with 0.25 D for 24 children remaining in glasses (p < 0.0001). Increased growth of the vitreous chamber appeared to account for much of this excess myopia progression, although the difference in that variable did not reach statistical significance (p = 0.101). We also noted a 0.203 D steepening in the corneal curvature in contact lens wearers compared with spectacle wearers whose corneas steepened very little (0.014 D, p = 0.007). Soft contact lens wear was also accompanied by a greater change in the near-point phoria which moved 4.5 prism dioptres in the exo direction compared with spectacle wearers who experienced only a 1.4 prism dioptre divergent shift (p = 0.048). PMID- 12535060 TI - The combined influence of knowledge, training and experience when grading contact lens complications. AB - PURPOSE: A study was conducted to evaluate the influence of knowledge, training and experience (clinical skills set) when assessing the severity of contact lens complications. METHODS: Nine optometrists (who were in possession of a relevant clinical skills set) and nine 'non-optometrists' (subjects without the clinical skills set) were each invited to grade - to the nearest 0.1 increment - an image of each of 16 contact lens complications using Efron Grading Scales for Contact Lens Complications. This procedure was repeated 2 weeks later, yielding a total data base comprising 576 individual grading estimates. RESULTS: The mean of the test and retest grading estimates was the same for the optometrists (2.8 +/- 0.7) and the non-optometrists (2.6 +/- 0.9) (F1,15,1 = 1.3, p = 0.26); that is, non optometrists can grade accurately. Median grading reliability for optometrists (+/-0.41) was lower than (i.e. superior to) that for non-optometrists (+/-0.67) (p = 0.001). Non-optometrists tended to display a reluctance to grade by interpolation and to less reliably grade subtle clinical signs. CONCLUSIONS: When averaged over several attempts, non-optometrists will arrive at similar estimates of severity to optometrists when grading ocular complications of contact lens wear; however, they will do so less reliably. The relative contribution of the three attributes of the clinical skills set to grading performance is presently unclear. PMID- 12535061 TI - Relationship between goblet cell density and tear function tests. AB - AIM: The objective of this study was to find a relationship between goblet cell density (GCD) and other diagnostic tests of dry eye in a group of normal healthy Chinese subjects. The capability of using GCD as a tear function test was assessed. METHOD: A total of 42 optometry students with no contact lens wear history were recruited. Subjects were required to fill in a dry eye questionnaire. The non-invasive tear break-up time (NITBUT) of each subject was measured, followed by phenol red thread test (PRTT) and tear break-up time (TBUT) tests. Conjunctival epithelial cells from the inferior nasal bulbar conjunctiva were harvested by the impression cytology technique. The specimens collected were labelled and stained with PAS (Periodic Acid Schiff)-haematoxylin. The goblet cells and the conjunctival epithelial cells were counted under a light microscope of 100x magnification. Spearman's rho (rho) correlation test was used to analyse the relationship between GCD and the other tear function tests. RESULTS: With a probability level of 0.0051 (corrected for a cumulative familywise error rate of p = 0.05), GCD was found to have no correlation with NITBUT (rho = -0.193, p = 0.222), McMonnies Dry Eye Scores (MCDES) (rho = -0.052, p = 0.742), PRTT (rho = - 0.188, p = 0.234) and TBUT (rho = 0.246, p = 0.117). CONCLUSION: There was no correlation between GCD and MCDES, NITBUT, PRTT and TBUT. The GCD alone is not a useful diagnostic test for tear film instability in normal eyes although it is a good indicator in the diagnosis of ocular surface diseases. PMID- 12535063 TI - EspH, a new cytoskeleton-modulating effector of enterohaemorrhagic and enteropathogenic Escherichia coli. AB - Enteropathogenic Escherichia coli (EPEC) and enterohaemorrhagic E. coli (EHEC) are closely related pathogens. During infection, EPEC and EHEC use a type III secretion system (TTSS) to translocate effector proteins into the infected cells and thereby modify specific host functions. These include transient filopodium formation which is Cdc42-dependent. Filopodia formation is followed by assembly of actin pedestals, the process enhanced by inhibition of Cdc42. We discovered that orf 18 of the enterocyte effacement locus encodes a new effector, which we termed EspH. We show that EspH is translocated efficiently into the infected cells by the TTSS and localizes beneath the EPEC microcolonies. Inactivation of espH resulted in enhanced formation of filopodia and attenuated the pedestals formation. Furthermore, overexpression of EspH resulted in strong repression of filopodium formation and heightened pedestal formation. We also demonstrate that overexpression of EspH by EHEC induces marked elongation of the typically flat pedestals. Similar pedestal elongation was seen upon infection of COS cells overexpressing EspH. EspH transiently expressed by the COS cells was localized to the membrane and disrupted the actin cytoskeletal structure. Our findings indicate that EspH is a modulator of the host actin cytoskeleton structure. PMID- 12535064 TI - Human milk lactoferrin is a serine protease that cleaves Haemophilus surface proteins at arginine-rich sites. AB - Lactoferrin is a member of the lactotransferrin family of non-haem, iron-binding glycoproteins and is found at high concentrations in all human secretions, where it plays a major role in mucosal defence. In recent work, we observed that lactoferrin has proteolytic activity and attenuates the pathogenic potential of Haemophilus influenzae by cleaving and removing two putative colonization factors, namely the IgA1 protease protein and the Hap adhesin. Experiments with protease inhibitors further suggested that lactoferrin may belong to a serine protease family. In the present study we explored the mechanism of lactoferrin protease activity and discovered that mutation of either Ser259 or Lys73 results in a dramatic decrease in proteolysis. Examination of the crystal structure revealed that these two residues are located in the N-terminal lobe of the protein, adjacent to a 12-15 A cleft that separates the N-lobe and the C-lobe and that can readily accommodate large polypeptide substrates. In additional work, we found that lactoferrin cleaves IgA1 protease at an arginine-rich region defined by amino acids 1379-1386 (RRSRRSVR) and digests Hap at an arginine-rich sequence between amino acids 1016 and 1023 (VRSRRAAR). Based on our results, we conclude that lactoferrin is a serine protease capable of cleaving arginine-rich sequences. We speculate that Ser259 and Lys73 form a catalytic dyad, reminiscent of a number of bacterial serine proteases. In addition, we speculate that lactoferrin may cleave arginine-rich sequences in a variety of microbial virulence proteins, contributing to its long-recognized antimicrobial properties. PMID- 12535065 TI - Lack of SeqA focus formation, specific DNA binding and proper protein multimerization in the Escherichia coli sequestration mutant seqA2. AB - In Escherichia coli wild-type cells newly formed origins cannot be reinitiated. The prevention of reinitiation is termed sequestration and is dependent on the hemimethylated state of newly replicated DNA. Several mutants discovered in a screen for the inability to sequester hemimethylated origins have been mapped to the seqA gene. Here, one of these mutants, seqA2, harbouring a single amino acid change in the C-terminal end of the SeqA protein, was found to also be unable to form foci in vivo. The SeqA foci seen in the wild-type cells are believed to arise from multimerization of SeqA on hemimethylated DNA at the replication fork, presumably representing organization of newly formed DNA by SeqA. The result suggests that the process of origin sequestration is closely tied to the process of focus maintenance at the replication fork. In vitro, purified SeqA2 protein was found incapable of forming highly ordered multimers that bind hemimethylated oriC. The mutant protein was also incapable of restraining negative supercoils. Both in vivo and in vitro results support the idea that origin sequestration is an integral part of organization of newly formed DNA performed by SeqA. PMID- 12535066 TI - Conservation of a gene conversion mechanism in two distantly related paralogues of Anaplasma marginale. AB - Anaplasmataceae, the causative agents of anaplasmosis and ehrlichiosis, persist in the bloodstream of their mammalian hosts, allowing acquisition and transmission by tick vectors. Anaplasma marginale establishes persistent infection characterized by sequential cycles of rickettsaemia in which new antigenic variants emerge. The two most immunodominant outer membrane proteins, MSP2 and MSP3, are paralogues, each encoded by a distinct family of related genes. This study demonstrates that, although the two gene families have diverged substantially, each has maintained a similar mechanism to generate structurally and antigenically polymorphic surface antigens. Like MSP2, MSP3 is expressed from a single locus in which variation of the expressed msp3 gene is generated by recombination using msp3 pseudogenes. Each of the msp3 pseudogenes encodes a unique central variable region (CVR) flanked by conserved 5' and 3' regions. Changes in the CVR of the expressed msp3, concomitant with invariance of the pseudogenes, indicate that expression site variation is generated using gene conversion. A. marginale thus maintains two large, separate systems within its small genome to generate antigenic variation of its surface proteins, while analogous structural elements indicate a common mechanism. PMID- 12535067 TI - A missense mutation in ftsZ differentially affects vegetative and developmentally controlled cell division in Streptomyces coelicolor A3(2). AB - Streptomyces coelicolor A3(2) undergoes at least two kinds of cell division: vegetative septation leading to cross-walls in the substrate mycelium; and developmentally regulated sporulation septation in aerial hyphae. By isolation and characterization of a non-sporulating ftsZ mutant, we demonstrate a difference between the two types of septation. The ftsZ17(Spo) allele gave rise to a classical white phenotype. The mutant grew as well as the parent on plates, and formed apparently normal hyphal cross-walls, although with a small reduction in frequency. In contrast, sporulation septation was almost completely abolished, resulting in a phenotype reminiscent of whiH and ftsZdelta2p mutants. The ftsZ17(Spo) allele was partially dominant and had no detectable effect on the cellular FtsZ content. As judged from both immunofluorescence microscopy of FtsZ and translational fusion of ftsZ to egfp, the mutation prevented correct temporal and spatial assembly of Z rings in sporulating hyphae. Homology modelling of S. coelicolor FtsZ indicated that the mutation, an A249T change in the C-terminal domain, would be expected to alter the protein on the lateral face of FtsZ protofilaments. The results suggest that cytokinesis may be developmentally controlled at the level of Z-ring assembly during sporulation of S. coelicolor A3(2). PMID- 12535068 TI - Motility modes of Spiroplasma melliferum BC3: a helical, wall-less bacterium driven by a linear motor. AB - Spiroplasma are members of the Mollicutes (Mycoplasma, Acholeplasma and Spiroplasma) - the simplest, minimal, free-living and self-replicating forms of life. The mollicutes are unique among bacteria in completely lacking cell walls and flagella and in having an internal, contractile cytoskeleton, which also functions as a linear motor. Spiroplasma are helical, chemotactic and viscotactic active swimmers. The Spiroplasmal cytoskeleton is a flat ribbon composed of seven pairs of fibrils. The ribbon is attached to the inner side of the cell membrane along its innermost (shortest) helical line. The cell's geometry and dynamic helical parameters, and consequently motility, can be controlled by changing differentially and in a co-ordinated manner, the length of the fibrils. We identified several consistent modes of cell movements and motility originating, most likely, as a result of co-operative or local molecular switching of fibrils: (i). regular extension and contraction within the limits of helical symmetry (this mode also includes straightening, beyond what is allowed by helical symmetry, and reversible change of helical sense); (ii). spontaneous and random change of helical sense originating at random sites along the cell (these changes propagate along the cell in either direction and hand switching is completed within approximately 0.08 second); (iii). forming a deformation on one of the helical turns and propagating it along the cell (these helical deformations may travel along the cell at a speed of up to approximately 40 microm s-1); (iv). random bending, flexing and twitching (equivalent to tumbling). In standard medium (viscosity = 1.147 centipoise) the cells run at approximately 1.5 microm s 1, have a Reynolds number of approximately 3.5 x 10-6 and consume approximately 30 ATP molecules s-1. Running velocity, duration, persistence and efficiency increase with viscosity upon adding ficoll, dextran and methylcellulose to standard media. Relative force measurements using optical tweezers confirm these findings. PMID- 12535069 TI - The bacterial linear motor of Spiroplasma melliferum BC3: from single molecules to swimming cells. AB - Spiroplasma melliferum BC3 are wall-less bacteria with internal cytoskeletons. Spiroplasma, Mycoplasma and Acholeplasma belong to the Mollicutes, which represent the smallest, simplest and minimal free-living and self-replicating forms of life. The Mollicutes are motile and chemotactic. Spiroplasma cells are, in addition, helical in shape. Based on data merging, obtained by video dark field light microscopy of live, swimming helical Spiroplasma cells and by cryoelectron microscopy, unravelling the subcellular structure and molecular organization of the cytoskeleton, we propose a functional model in which the cytoskeleton also acts as a bacterial linear motor enabling and controlling both dynamic helicity and swimming. The cytoskeleton is a flat, monolayered ribbon constructed from seven contractile fibrils (generators) capable of changing their length differentially in a co-ordinated manner. The individual, flat, paired fibrils can be viewed as chains of tetramers approximately 100 A in diameter composed of 59 kDa monomers. The cytoskeletal ribbon is attached to the inner surface of the cell membrane (but is not an integral part of it) and follows the shortest helical line on the coiled cellular tube. We show that Spiroplasma cells can be regarded, at least in some states, as near-perfect dynamic helical tubes. Thus, the analysis of experimental data is reduced to a geometrical problem. The proposed model is based on simple structural elements and functional assumptions: rigid circular rings are threaded on a flexible, helical centreline. The rings maintain their circularity and normality to the centreline at all helical states. An array of peripheral, equidistant axial lines forms a regular cylindrical grid (membrane), by crossing the lines bounding the rings. The axial and peripheral spacing correspond to the tetramer diameter and fibril width (100 A) respectively. Based on electron microscopy data, we assign seven of the axial grid lines in the model to function as contractile generators. The generators are clustered along the shortest helical paths on the cellular coil. In the model, the shortest generator coincides with the shortest helical line. The rest, progressively longer, six generators follow to the right or to the left of the shortest generator in order to generate the maximal range of lengths. A rubbery membrane is stretched over (or represented by) the three-dimensional grid to form a continuous tube. Co-ordinated, differential length changes of the generators induce the membranal cylinder to coil and uncoil reversibly. The switch of helical sense requires equalization of the generators' length, forming a straight cylindrical tube with straight generators. The helical parameters of the cell population, obtained by light microscopy, constitute several subpopulations related, most probably, to cell size and age. The range of molecular dimensions in the active cytoskeleton inferred from light microscopy and modelling agrees with data obtained by direct measurements of subunit images on electron micrographs, scanning transmission electron microscopy (STEM) and diffraction analysis of isolated ribbons. Swimming motility and chemotactic responses are carried out by one or a combination of the following: (i). reciprocating helical extension and compression ('breathing'); (ii). propagation of a deformation (kink) along the helical path; (iii). propagation of a reversal of the helical sense along the cell body; and (iv). irregular flexing and twitching, which is functionally equivalent to standard bacterial tumbling. Here, we analyse in detail only the first case (from which all the rest are derived), including switching of the helical sense. PMID- 12535070 TI - Specialized osmotic stress response systems involve multiple SigB-like sigma factors in Streptomyces coelicolor. AB - Whereas in Bacillus subtilis, a general stress response stimulon under the control of a single sigma factor (SigB) is induced by different physiological and environmental stresses (heat, salt or ethanol shock), in Streptomyces coelicolor, these environmental stresses induce independent sets of proteins, and its genome encodes nine SigB paralogues. To investigate possible functions of multiple sigB like genes in S. coelicolor, Pctc, a promoter routinely used to assay SigB activity in vivo, was analysed as a heterologous reporter. The fact that Pctc was activated by osmotic shock, but not by heat or ethanol, confirmed that stress response system(s) could operate independently in S. coelicolor. Pctc was also induced transiently during growth of liquid cultures, presumably by nutritional signals. We purified an RNA polymerase holoenzyme from crude extracts that catalysed specific transcription of Pctc in vitro. Its sigma subunit was identified as a product of the sigH gene, which is co-transcribed downstream of a putative antisigma factor gene (prsH). Although the sigH function was not needed for normal colony morphology, prsH was conditionally required for both aerial hyphae formation and regulation of antibiotic biosynthesis. Levels of two different sigH-encoded proteins were growth phase dependent but not significantly changed by osmotic stress, implying the important roles of post-translational regulatory elements such as PrsH. In addition, synthesis of three other SigH-like proteins was induced by osmotic stress, but not by ethanol or heat. Two of them were genetically assigned to sigH homologous loci sigI and sigJ and shown to be independently regulated. This family of SigH-like proteins displayed different osmotic response kinetics. Thus, in contrast to many other bacteria, S. coelicolor uses an osmotic sensory system that can co-ordinate the activity of multiple paralogues to control the relative activity of promoters within the same stress stimulon. Such specialized stress response systems may reflect adaptive functions needed for colonial differentiation. PMID- 12535071 TI - AraC/XylS family members, HilD and HilC, directly activate virulence gene expression independently of HilA in Salmonella typhimurium. AB - Salmonella typhimurium is a Gram-negative enteric pathogen that can infect intestinal epithelial cells and induce inflammation of the intestinal mucosa. These processes are mediated by a type III secretion system (TTSS), which is encoded on Salmonella pathogenicity island 1 (SPI1). Previous studies showed that four SPI1-encoded transcriptional regulators, HilD, HilC, HilA and InvF, act in an ordered fashion to co-ordinately activate expression of the SPI1 TTSS. HilD and HilC derepress hilA transcription. HilA activates invF as well as SPI1 genes that encode components of the TTS apparatus. InvF then activates genes that encode proteins secreted by the SPI1 TTS apparatus. In this scheme, HilD and HilC indirectly activate expression of the SPI1 TTS apparatus and its secreted substrates by affecting hilA expression. Here, we report that HilD and HilC can also activate expression of a subset of SPI1 genes independently of HilA. Our studies show that HilD and HilC activate transcription of invF from a promoter that is far upstream of its HilA-dependent promoter. This activation is most probably through direct binding of HilD and HilC to sequences upstream and downstream of this alternative HilA-independent promoter. We conclude that HilD and HilC have a second role in SPI1 gene regulation that is separate from their role in co-ordinating expression of the SPI1 TTSS through hilA. PMID- 12535072 TI - The crk1 gene encodes an Ime2-related protein that is required for morphogenesis in the plant pathogen Ustilago maydis. AB - The fungal pathogen Ustilago maydis alternates between budding and filamentous growth during its life cycle. This dimorphic transition is regulated by environmental factors and mating. We cloned a new gene, crk1, which encodes a protein with sequence similarity to Ime2, a kinase involved in developmental choices in S. cerevisiae. Disruption of the crk1 gene in U. maydis resulted in cells that are unable to respond in an appropriate manner to environmental stimuli and show defects in morphogenesis and cell cycle adjustment to changing conditions. We have analysed the regulation of the crk1 gene and demonstrated that cAMP and MAPK pathways have opposite influences on the transcript levels for crk1. Furthermore, we have shown that alterations in the components of these pathways impair the ability of the cellular machinery to adapt to changing conditions. These results demonstrate an important role for the crk1- encoded protein in the morphogenesis and environmental adaptation in Ustilago maydis. PMID- 12535073 TI - Domestication of wild Saccharomyces cerevisiae is accompanied by changes in gene expression and colony morphology. AB - Although colonies from Saccharomyces cerevisiae laboratory strains are smooth, those isolated from nature exhibit a structured fluffy pattern. Environmental scanning electron microscopy shows that the cells within wild fluffy colonies are connected by extracellular matrix (ECM) material. This material contains a protein of about 200 kDa unrelated to the flocculins, proteins involved in cell cell adhesion in liquid media. The matrix material binds to concanavalin A. Within a few passages on rich agar medium, the wild strains switch from the fluffy to the smooth colony morphology. This domestication is accompanied by loss of the ECM and by extensive changes in gene expression as detected by DNA microarrays. The expression of about 320 genes was changed in smooth colonies. The major changes comprise carbohydrate metabolism, cell wall, water channels, Ty transposons and subtelomeric genes, iron homeostasis, vitamin metabolism and cell cycle and polarity. The growth in fluffy colonies may represent a metabolic strategy for survival of yeast under unfavourable conditions that is switched off under felicitous laboratory conditions. PMID- 12535074 TI - The DnaAcos allele of Escherichia coli: hyperactive initiation is caused by substitution of A184V and Y271H, resulting in defective ATP binding and aberrant DNA replication control. AB - Chromosomal DNA replication is regulated at the level of commitment to this biochemical pathway. In Escherichia coli, DnaA protein appears to regulate this process. A mutant form, DnaAcos, carrying four amino acid substitutions, is apparently defective in responding to regulatory signals, because it induces hyperactive initiation from the bacterial replication origin (oriC). In this report, the phenotype of hyperactive initiation is shown to be the result of two specific amino acid substitutions. One (A184V) immediately adjacent to a Walker A box (P loop motif) causes a defect in ATP binding (Carr and Kaguni, 1996, Mol Microbiol 20: 1307-1318). The second amino acid substitution (Y271H) appears to stabilize the activity of the mutant protein carrying the A184V substitution. The mutant protein carrying both amino acid substitutions (A184V + Y271H) is defective in modulating the frequency of initiation from oriC, as demonstrated by marker frequency analysis of oriC and a locus near the replication terminus. These results indicate that a defect in ATP binding results in aberrant control of DNA replication. PMID- 12535075 TI - Characterization of potassium transport in wild-type and isogenic yeast strains carrying all combinations of trk1, trk2 and tok1 null mutations. AB - Saccharomyces cerevisiae cells express three defined potassium-specific transport systems en-coded by TRK1, TRK2 and TOK1. To gain a more complete understanding of the physiological function of these transport proteins, we have constructed a set of isogenic yeast strains carrying all combinations of trk1delta, trk2delta and tok1delta null mutations. The in vivo K+ transport characteristics of each strain have been documented using growth-based assays, and the in vitro biochemical and electrophysiological properties associated with K+ transport have been determined. As has been reported previously, Trk1p and Trk2p facilitate high affinity potassium uptake and appear to be functionally redundant under a wide range of environmental conditions. In the absence of TRK1 and TRK2, strains lack the ability specifically to take up K+, and trk1deltatrk2delta double mutant cells depend upon poorly understood non-specific cation uptake mechanisms for growth. Under conditions that impair the activity of the non-specific uptake system, termed NSC1, we have found that the presence of functional Tok1p renders cells sensitive to Cs+. Based on this finding, we have established a growth-based assay that monitors the in vivo activity of Tok1p. PMID- 12535076 TI - Modified yeast cells to investigate the coupling of G protein-coupled receptors to specific G proteins. AB - G protein-coupled receptors (GPCRs) help to regulate the physiology of all the major organ systems. They respond to a multitude of ligands and activate a range of effector proteins to bring about the appropriate cellular response. The choice of effector is largely determined by the interaction of individual GPCRs with different G proteins. Several factors influence this interaction, and a better understanding of the process may enable a more rational approach to identifying compounds that affect particular signalling pathways. A number of systems have been developed for the analysis of GPCRs. All provide useful information, but the genetic amenability and relative simplicity of yeast makes them a particularly attractive option for ligand identification and pharmaceutical screening. Many, but not all, GPCRs are functional in the budding yeast Saccharomyces cerevisiae, and we have developed reporter strains of the fission yeast Schizosaccharomyces pombe as an alternative host. To provide a more generic system for investigating GPCRs, we created a series of yeast-human Galpha-transplants, in which the last five residues at the C-terminus of the yeast Galpha-subunit are replaced with the corresponding residues from different human G proteins. These enable GPCRs to be coupled to the Sz. pombe signalling machinery so that stimulation with an appropriate ligand induces the expression of a signal-dependent lacZ reporter gene. We demonstrate the specificity of the system using corticotropin releasing factor (CRF) and CRF-related peptides on two CRF receptors. We find that different combinations of ligand and receptor activate different Galpha transplants, and the specificity of the coupling is similar to that in mammalian systems. Thus, CRF signalled through the Gs- and Gi-transplants, consistent with its regulation of adenylate cyclase, and was more active against the CRF-R1A receptor than against the CRF-R2B receptor. In contrast, urocortin II and urocortin III were selective for the CRF-R2B receptors. Furthermore, urocortin, but not CRF, induced signalling through the CRF-R1A receptor and the Gq transplant. This is the first time that human GPCRs have been coupled to the signalling pathway in Sz. pombe, and the strains described in this study will complement the other systems available for studying this important family of receptors. PMID- 12535077 TI - A new Escherichia coli metabolic competency: growth on fatty acids by a novel anaerobic beta-oxidation pathway. AB - Escherichia coli uses fatty acids as a sole carbon and energy source during aerobic growth by means of the enzymes encoded by the fad regulon. We report that this bacterium can also grow on fatty acids under anaerobic conditions provided that a terminal respiratory electron acceptor such as nitrate is available. This anaerobic utilization pathway is distinct from the well-studied aerobic pathway in that (i). it proceeds normally in mutant strains lacking various enzymes of the aerobic pathway; (ii). it functions with fatty acids (octanoate and decanoate) that cannot be used by wild-type E. coli strains under aerobic conditions; and (iii). super-repressor mutants of the fadR regulatory locus that block aerobic growth on fatty acids fail to block the anaerobic pathway. We have identified homologues of the FadA, FadB and FadD proteins required for aerobic fatty acid utilization called YfcY, YfcX and YdiD, respectively, which are involved in anaerobic growth on fatty acids. A strong FadR binding site was detected upstream of the yfcY gene consistent with microarray analyses, indicating that yfcYX expression is negatively regulated by FadR under aerobic growth conditions. In contrast, transcriptional regulation of ydiD appears to be independent of FadR, and anaerobic growth on fatty acids is not under FadR control. These three genes are conserved in the available genome sequences of pathogenic E. coli, Shigella and Salmonella strains. PMID- 12535078 TI - Translocation of YopE and YopN into eukaryotic cells by Yersinia pestis yopN, tyeA, sycN, yscB and lcrG deletion mutants measured using a phosphorylatable peptide tag and phosphospecific antibodies. AB - Yersinia pestis, the causative agent of plague, exports a set of virulence proteins called Yops upon contact with eukaryotic cells. A subset of these Yops is translocated directly into the cytosol of host cells. In this study, a novel protein tag-based reporter system is used to measure the translocation of Yops into cultured eukaryotic cells. The reporter system uses a small bipartite phosphorylatable peptide tag, termed the Elk tag. Translocation of an Elk-tagged protein into eukaryotic cells results in host cell protein kinase-dependent phosphorylation of the tag at a specific serine residue, which can subsequently be detected with phosphospecific antibodies. The YopN, TyeA, SycN, YscB and LcrG proteins function to prevent Yop secretion before host cell contact. The role of these proteins was investigated in the translocation of Elk-tagged YopE (YopE129 Elk) and YopN (YopN293-Elk) into HeLa cells. Y. pestis yopN, tyeA, sycN and yscB deletion mutants showed reduced levels of YopE129-Elk phosphorylation compared with the parent strain, indicating that these mutants translocate reduced amounts of YopE. We also demonstrate that YopN293-Elk is translocated into HeLa cells and that this process is more efficient in a Yersinia yop polymutant strain lacking the six translocated effector Yops. Y. pestis sycN and yscB mutants translocated reduced amounts of YopN293-Elk; however, tyeA and lcrG mutants translocated higher amounts of YopN293-Elk compared with the parent strain. These data suggest that TyeA and LcrG function to suppress the secretion of YopN before host cell contact, whereas SycN and YscB facilitate YopN secretion and subsequent translocation. PMID- 12535079 TI - An extended role for the nucleoid structuring protein H-NS in the virulence gene regulatory cascade of Shigella flexneri. AB - The H-NS nucleoid structuring protein has been shown previously to play a negative role in controlling virulence gene expression in Shigella flexneri by repressing transcription of the virF and virB regulatory genes and the VirF dependent icsA structural gene under non-permissive growth conditions. Here, we show that H-NS also acts at the promoters of the VirB-dependent structural genes in the regulatory cascade. H-NS protein binds to the promoter regions in vivo and in vitro. The promoters were shown physically and by in silico analysis to contain regions of DNA curvature, a feature of H-NS binding sites. H-NS binding sites were determined by DNase I footprinting at the icsB and the virA promoters. The locations of these sites were consistent with a role for H-NS as a transcription repressor. The VirB-dependent structural gene promoters were found to respond directly to the H-NS repressor, revealing a level of control that is additional to that exerted by the H-NS-dependent virB activator gene. Moreover, the promoters were sensitive to the level of VirB protein in the cell, requiring a threshold level of VirB to be reached before becoming active. A model is discussed in which the levels of expression of the structural genes reflect the outcome of competition between the countervailing regulatory activities of the H NS and VirB proteins. PMID- 12535080 TI - The transmembrane domains of the sensor kinase KdpD of Escherichia coli are not essential for sensing K+ limitation. AB - The sensor kinase/response regulator system KdpD/KdpE of Escherichia coli regulates the expression of the kdpFABC operon, which encodes the high affinity K+ transport system KdpFABC. The membrane-bound sensor kinase KdpD consists of four transmembrane domains, a large cytoplasmic N-terminal domain and a cytoplasmic C-terminal transmitter domain. To elucidate the role of the four transmembrane domains, various deletions were introduced in kdpD and the activities of the resulting truncated derivatives of KdpD were determined. A KdpD protein lacking all four transmembrane domains was able to sense low K+ concentrations, whereas at higher K+ concentrations kdpFABC expression was constitutive. These and further results with various truncated KdpD proteins lacking distinct parts of the transmembrane domains or derivatives in which a linker peptide or two transmembrane domains of PutP, the Na+/proline transporter of Escherichia coli, replaced the missing part indicated that the transmembrane domains are not essential for sensing of K+ limitation, but may be important for the correct positioning of the large N- and C-terminal cytoplasmic domains to each other. PMID- 12535081 TI - Acyl-homoserine lactone acylase from Ralstonia strain XJ12B represents a novel and potent class of quorum-quenching enzymes. AB - N-acylhomoserine lactones (AHLs) are used as signal molecules by many quorum sensing Proteobacteria. Diverse plant and animal pathogens use AHLs to regulate infection and virulence functions. These signals are subject to biological inactivation by AHL-lactonases and AHL-acylases. Previously, little was known about the molecular details underlying the latter mechanism. An AHL signal inactivating bacterium, identified as a Ralstonia sp., was isolated from a mixed species biofilm. The signal inactivation encoding gene from this organism, which we call aiiD, was cloned and successfully expressed in Escherichia coli and inactivated three AHLs tested. The predicted 794-amino-acid polypeptide was most similar to the aculeacin A acylase (AAC) from Actinoplanes utahensis and also shared significant similarities with cephalosporin acylases and other N-terminal (Ntn) hydrolases. However, the most similar homologues of AiiD are deduced proteins of undemonstrated function from available Ralstonia, Deinococcus and Pseudomonas genomes. LC-MS analyses demonstrated that AiiD hydrolyses the AHL amide, releasing homoserine lactone and the corresponding fatty acid. Expression of AiiD in Pseudomonas aeruginosa PAO1 quenched quorum sensing by this bacterium, decreasing its ability to swarm, produce elastase and pyocyanin and to paralyze nematodes. Thus, AHL-acylases have fundamental implications and hold biotechnological promise in quenching quorum sensing. PMID- 12535082 TI - Streptococcus pyogenes recruits collagen via surface-bound fibronectin: a novel colonization and immune evasion mechanism. AB - This study aimed to characterize matrix assembly mechanisms on the surface of the human pathogen Streptococcus pyogenes. Among 125 S. pyogenes isolates, 61% were able to recruit collagen type IV via surface-bound fibronectin. Streptococcus gordonii expressing the fibronectin-binding repeat domain of S. pyogenes SfbI protein was equally potent in recruiting collagen, indicating that this domain was sufficient to promote fibronectin-mediated collagen recruitment. Electron microscopic analysis of streptococci revealed that fibronectin-mediated collagen recruitment led to matrix deposition on and between streptococcal cells, which induced the formation of large bacterial aggregates. Furthermore, collagen recruiting streptococci were able to colonize collagen fibres and were protected from adhering to human polymorphonuclear cells in the presence of opsonizing antibodies. Fibronectin-mediated collagen recruitment thus represents a novel aggregation, colonization and immune evasion mechanism of S. pyogenes. PMID- 12535083 TI - History and evolution of the arctic flora: in the footsteps of Eric Hulten. AB - A major contribution to our initial understanding of the origin, history and biogeography of the present-day arctic flora was made by Eric Hulten in his landmark book Outline of the History of Arctic and Boreal Biota during the Quarternary Period, published in 1937. Here we review recent molecular and fossil evidence that has tested some of Hulten's proposals. There is now excellent fossil, molecular and phytogeographical evidence to support Hulten's proposal that Beringia was a major northern refugium for arctic plants throughout the Quaternary. In contrast, most molecular evidence fails to support his proposal that contemporary east and west Atlantic populations of circumarctic and amphi Atlantic species have been separated throughout the Quaternary. In fact, populations of these species from opposite sides of the Atlantic are normally genetically very similar, thus the North Atlantic does not appear to have been a strong barrier to their dispersal during the Quaternary. Hulten made no detailed proposals on mechanisms of speciation in the Arctic; however, molecular studies have confirmed that many arctic plants are allopolyploid, and some of them most probably originated during the Holocene. Recurrent formation of polyploids from differentiated diploid or more low-ploid populations provides one explanation for the intriguing taxonomic complexity of the arctic flora, also noted by Hulten. In addition, population fragmentation during glacial periods may have lead to the formation of new sibling species at the diploid level. Despite the progress made since Hulten wrote his book, there remain large gaps in our knowledge of the history of the arctic flora, especially about the origins of the founding stocks of this flora which first appeared in the Arctic at the end of the Pliocene (approximately 3 Ma). Comprehensive analyses of the molecular phylogeography of arctic taxa and their relatives together with detailed fossil studies are required to fill these gaps. PMID- 12535084 TI - Divergent selection maintains adaptive differentiation despite high gene flow between sympatric rainbow smelt ecotypes (Osmerus mordax Mitchill). AB - In this study, we investigate the relative role of historical factors and evolutionary forces in promoting population differentiation in a new case of sympatric dwarf and normal ecotypes of the rainbow smelt (Osmerus mordax Mitchill) in Lac Saint-Jean (Quebec, Canada). Our first objective was to test the hypothesis that the evolution of sympatric smelt ecotypes in Lac Saint-Jean has been contingent upon the secondary contact between two evolutionary lineages in postglacial times. Secondly, the QST method was applied to test the null hypothesis that the extent of phenotypic differences relative to that of neutral marker variation would be similar in comparisons involving populations within and among ecotypes. Thirdly, we applied a quantitative-genetic method as an exploratory assessment as to whether the amount of gene flow observed between populations could affect divergence in adaptive traits under specific conditions. This study revealed a unique situation of dwarf and normal smelt ecotypes that are, respectively, characterized by selmiparous and iteroparous life histories and the occurrence in each of two genetically distinct populations that synchronously use the same spawning habitat in two tributaries. Historical contingency has apparently played little role in the origin of these populations. In contrast, an important role of divergent natural selection in driving their phenotypic divergence was suggested. While divergent selection has apparently been strong enough to maintain phenotypic differentiation in the face of migration, this study suggests that gene flow has been sufficiently important to modulate the extent of adaptive differentiation being achieved between ecotypes, unless the extent of stabilizing selection acting on smelt ecotypes is much more pronounced than usually reported in natural populations. PMID- 12535085 TI - Population genetic consequences of extreme variation in sexual and clonal reproduction in an aquatic plant. AB - Most plants combine sexual reproduction with asexual clonal reproduction in varying degrees, yet the genetic consequences of reproductive variation remain poorly understood. The aquatic plant Butomus umbellatus exhibits striking reproductive variation related to ploidy. Diploids produce abundant viable seed whereas triploids are sexually sterile. Diploids also produce hundreds of tiny clonal bulbils, whereas triploids exhibit only limited clonal multiplication through rhizome fragmentation. We investigated whether this marked difference in reproductive strategy influences the diversity of genotypes within populations and their movement between populations by performing two large-scale population surveys (n = 58 populations) and assaying genotypic variation using random amplified polymorphic DNA (RAPDs). Contrary to expectations, sexually fertile populations did not exhibit higher genotypic diversity than sterile populations. For each cytotype, we detected one very common and widespread genotype. This would only occur with a very low probability (< 10-7) under regular sexual recombination. Compatibility analysis also indicated that the pattern of genotypic variation largely conformed to that expected with predominant clonal reproduction. The potential for recombination in diploids is not realized, possibly because seeds are outcompeted by bulbils for safe sites during establishment. We also failed to find evidence for more extensive movement of fertile than sterile genotypes. Aside from the few widespread genotypes, most were restricted to single populations. Genotypes in fertile populations were very strongly differentiated from those in sterile populations, suggesting that new triploids have not arisen during the colonization of North America. The colonization of North America involves two distinct forms of B. umbellatus that, despite striking reproductive differences, exhibit largely clonal population genetic structures. PMID- 12535086 TI - Lack of genetic and plumage differentiation in the red-billed quelea Quelea quelea across a migratory divide in southern Africa. AB - A migratory divide usually signals the presence of a geographical region over which other traits, such as morphology and genotypes, also undergo rapid change. A migratory divide has been hypothesized in central southern Africa for the abundant migratory weaver, the red-billed quelea Quelea quelea. The positioning of the divide in the region is based on the patterns of rainfall in the region that stimulate the annual migrations of queleas. Evidence indicates that premigratory queleas near the divide show two distinct preferred directions for migration. We used eight polymorphic microsatellite loci and a range of plumage characters to determine whether there was population structure among red-billed queleas in southern Africa, and specifically whether this structure coincided with the location of the migratory divide. There was no evidence of population genetic structure. An amova revealed no significant differences between samples taken either side of the migratory divide. Similarly, there was no geographical variation in plumage patterns across southern Africa. For both microsatellites and plumage characteristics, the variation that does exist occurs within each sampled site, with little differentiation between sites. We were therefore unable to find any evidence that either plumage or microsatellite genotypes varied in a similar way to migratory direction preference in red-billed queleas in southern Africa. This is perhaps because the migratory divide does not act to separate individuals into populations within which genetic and plumage differentiation can be maintained. PMID- 12535087 TI - Extinction-colonization dynamics structure genetic variation of spotted sunfish (Lepomis punctatus) in the Florida Everglades. AB - The population genetics of aquatic animals in the Florida Everglades may be strongly influenced by extinction and colonization dynamics. We combined analyses of allozyme and microsatellite loci to test the hypothesis that two levels of population structure are present for spotted sunfish (Pisces: Centrarchidae: Lepomis punctatus) inhabiting the Everglades. We hypothesized that annual cycles of marsh dry-down increase local-scale genetic variation through a process of local extinction and colonization; we hypothesized that barriers to gene flow by levee/canal systems create a second, regional level of genetic variation. In 1996 and 1997, we sampled spotted sunfish from 11 Everglades sites that were distributed in three regions separated by levees. We documented patterns of genetic variation at 7 polymorphic allozyme loci and 5 polymorphic microsatellite loci. Most genetic variation was present among local populations, according to both types of genetic markers. Furthermore, samples from marsh sites were heterogeneous, while those from canals were not. These data supported our hypothesis that dry-down events and local population dynamics in the marsh have a significant effect on population genetic structure of spotted sunfish. We found no support for our hypothesis that water-management structures superimpose a second level of genetic structure on this species, possibly because canals obscure historical structure by facilitating gene flow or because the complete canal system has been in place for fewer than 20 generations of this species. Our data suggests a continent-island (canal-marsh) structure of populations with high gene flow among regions and recurrent mixing in marshes from canal and creek habitats. PMID- 12535088 TI - DNA sequence variation in BpMADS2 gene in two populations of Betula pendula. AB - The PISTILLATA (PI) homologue, BpMADS2, was isolated from silver birch (Betula pendula Roth) and used to study nucleotide polymorphism. Two regions (together about 2450 bp) comprising mainly untranslated sequences were sequenced from 10 individuals from each of two populations in Finland. The nucleotide polymorphism was low in the BpMADS2 locus, especially in the coding region. The synonymous site overall nucleotide diversity (pis) was 0.0043 and the nonsynonymous nucleotide diversity (pia) was only 0.000052. For the whole region, the pi values for the two populations were 0.0039 and 0.0045, and for the coding regions, the pi values were only 0 and 0.00066 (for the corresponding coding regions of Arabidopsis thaliana PI world-wide pi was 0.0021). Estimates of pi or theta did not differ significantly between the two populations, and the two populations were not diverged from each other. Two classes of BpMADS2 alleles were present in both populations, suggesting that this gene exhibits allelic dimorphism. In addition to the nucleotide site variation, two microsatellites were also associated within the haplotypes. This allelic dimorphism might be the result of postglacial re-colonization partly from northwestern, partly from southeastern/eastern refugia. The sequence comparison detected five recombination events in the regions studied. The large number of microsatellites in all of the three introns studied suggests that BpMADS2 is a hotspot for microsatellite formation. PMID- 12535089 TI - Fine-scaled geographical population structuring in a highly mobile marine species: the Atlantic cod. AB - Compared with many terrestrial and freshwater environments, dispersal and interbreeding is generally much less restricted in the marine environment. We studied the tendency for a marine species, the Atlantic cod, to be sub-structured into genetically differentiated populations on a fine geographical scale. We selected a coastal area free of any obvious physical barriers and restricted sampling to a 300-km region, well within the dispersal ability of this species. Screening 10 polymorphic microsatellite loci in 6 samples we detected a weak, but consistent, differentiation at all 10 loci. The average FST over loci was small (0.0023) but highly significant statistically, demonstrating that genetically differentiated populations can arise and persist in the absence of physical barriers or great distance. We found no geographical pattern in the genetic differentiation and there was no apparent trend of isolation by distance along the coastline. These findings lend support to the notion that low levels of differentiation are due to passive transport of eggs or larvae by the ocean currents rather than to adult dispersal, the latter being strongly dependent on distance. PMID- 12535090 TI - Multilocus sequence typing suggests the chytrid pathogen of amphibians is a recently emerged clone. AB - Chytridiomycosis is a recently identified fungal disease associated with global population declines of frogs. Although the fungus, Batrachochytrium dendrobatidis, is considered an emerging pathogen, little is known about its population genetics, including the origin of the current epidemic and how this relates to the dispersal ability of the fungus. In this study, we use multilocus sequence typing to examine genetic diversity and relationships among 35 fungal strains from North America, Africa and Australia. Only five variable nucleotide positions were detected among 10 loci (5918 bp). This low level of genetic variation is consistent with the description of B. dendrobatidis as a recently emerged disease agent. Fixed (i.e. 100%) or nearly fixed frequencies of heterozygous genotypes at two loci suggested that B. dendrobatidis is diploid and primarily reproduces clonally. In contrast to the lack of nucleotide polymorphism, electrophoretic karyotyping of multiple strains demonstrated a number of chromosome length polymorphisms. PMID- 12535091 TI - The recent evolutionary origin of the phenotypically novel amphipod Hyalella montezuma offers an ecological explanation for morphological stasis in a closely allied species complex. AB - Numerous molecular studies have identified morphologically cryptic, freshwater invertebrate species, but have not suggested possible mechanisms for their phenotypic stasis. The amphipod crustacean genus Hyalella contains numerous morphologically cryptic species in the H. azteca complex, as well as a small number of morphologically very divergent, narrowly endemic taxa. One such taxon, Hyalella montezuma, is the sole planktonic filter-feeder within the North American amphipod fauna, and is known only from Montezuma Well, a fishless travertine spring mound in Arizona, USA. In this study, we conduct a phylogenetic analysis of mtDNA sequence data using likelihood, Bayesian and cladistic approaches to determine both the relationship of H. montezuma to the H. 'azteca' species complex, and to ascertain if its morphological and ecological differentiation have been comparatively recent. The results show that H. montezuma has a very close phylogenetic affiliation with one lineage in the H. azteca complex, indicating that its origin has been recent. We present evidence suggesting that fish predation is an important ecological factor, which constrains morphological and ecological diversification within the genus Hyalella, and that Montezuma Well has provided a relaxation on this constraint. PMID- 12535092 TI - Tracing the geographical origin of Megastigmus transvaalensis (Hymenoptera: Torymidae): an African wasp feeding on a South American plant in North America. AB - Determining the geographical origin of an introduced organism can be critical to understanding or managing a non-native species, but is often difficult when the organism is small or inconspicuous. We used a phylogeographical approach to identify the region of endemism and determine the geographical origin of world populations of the seed-feeding wasp Megastigmus transvaalensis (Hussey). This wasp feeds on African Rhus species and South American Schinus species in various locations around the world. Because it is present both in Africa and in South America, it is unclear whether the wasp was originally an African Rhus-feeder that has begun feeding on Schinus or a South American Schinus-feeder that has started feeding on Rhus. Phylogenetic analysis of 800 bp of mitochondrial cytochrome oxidase I sequence data found extensive variation and phylogeographical structure within African M. transvaalensis. Specimens from other locations around the world were all identical in COI sequence and were phylogenetically nested within the African samples. We conclude that M. transvaalensis was originally an African Rhus-feeder that readily attacks Schinus. We evaluate potential pathways of introduction of this wasp to the New World, and we discuss implications of our results for biocontrol efforts against invasive Schinus populations. PMID- 12535093 TI - Possible causes of morphological variation in an endemic Moroccan groundsel (Senecio leucanthemifolius var. casablancae): evidence from chloroplast DNA and random amplified polymorphic DNA markers. AB - Genetic variation was assessed in Senecio leucanthemifolius var. casablancae (Compositae), a Moroccan Atlantic coast endemic, in order to examine possible causes of atypical leaf morphology in three populations south of the known range. Evidence for introgression from S. glaucus ssp. coronopifolius and/or divergence was investigated with molecular markers. Both random amplified polymorphic DNA (RAPD) and chloroplast (cp) DNA restriction fragment length polymorphism (RFLP) differentiated the species well. Some evidence that hybridization may have occurred between the two species was provided by cpDNA markers. However, biparentally inherited RAPD markers failed to provide any support for the hypothesis that intermediate leaf morphologies in atypical populations arose through hybridization. Consequently, they are most likely to have arisen via divergence caused by drift and/or selection. Genetic distances among populations of S. leucanthemifolius were significant in all but one case. Isolation by distance was indicated by a significant positive correlation between genetic and geographical distances (r = 0.68, P = 0.01, Mantel test). These results suggest that long-distance achene dispersal is rare, despite the presence of a well developed pappus. The observed loss of pappus at achene maturity may explain this unexpected result. Due to the morphological distinction of var. casablancae from other varieties of S. leucanthemifolius, we suggest elevation to species rank and treatment of the atypical material at infraspecific rank. PMID- 12535094 TI - Large extent of mitochondrial DNA transfer from Oreochromis aureus to O. niloticus in West Africa. AB - Introgressive hybridization has an important evolutionary significance in terms of gene diversity and speciation. Among the major groups of vertebrates, fish show a strong propensity to hybridize. In order to highlight the possible occurrence of gene flow between two tilapia species, Oreochromis niloticus and O. aureus, a comparison of allozyme and mitochondrial DNA (mtDNA) polymorphism was performed on sympatric and allopatric populations of these two species. Nuclear data were congruent with the morphological identification of O. niloticus and O. aureus populations. In opposition, the mtDNA analysis resulted in two strictly differentiated groups which did not follow the morphological and nuclear DNA classification. The first group consisted of East African O. niloticus populations and the second included all the O. aureus populations and the West African O. niloticus populations. Moreover, in some cases, the same sequences were detected in both species. These data strongly support a differential introgression of mtDNA from O. aureus to O. niloticus involving all the West African area. This work points out the risk of misinterpretation of mtDNA or nuclear DNA data when only one single class of marker is used. PMID- 12535095 TI - Introgression patterns in the mosaic hybrid zone between Mytilus edulis and M. galloprovincialis. AB - Hybrid zones are fascinating systems to investigate the structure of genetic barriers. Marine hybrid zones deserve more investigation because of the generally high dispersion potential of planktonic larvae which allows migration on scales unrivalled by terrestrial species. Here we analyse the genetic structure of the mosaic hybrid zone between the marine mussels Mytilus edulis and M. galloprovincialis, using three length-polymorphic PCR loci as neutral and diagnostic markers on 32 samples along the Atlantic coast of Europe. Instead of a single genetic gradient from M. galloprovincialis on the Iberian Peninsula to M. edulis populations in the North Sea, three successive transitions were observed in France. From South to North, the frequency of alleles typical of M. galloprovincialis first decreases in the southern Bay of Biscay, remains low in Charente, then increases in South Brittany, remains high in most of Brittany, and finally decreases again in South Normandy. The two enclosed patches observed in the midst of the mosaic hybrid zone in Charente and Brittany, although predominantly M. edulis-like and M. galloprovincialis-like, respectively, are genetically original in two respects. First, considering only the various alleles typical of one species, the patches show differentiated frequencies compared to the reference external populations. Second, each patch is partly introgressed by alleles of the other species. When introgression is taken into account, linkage disequilibria appear close to their maximum possible values, indicating a strong genetic barrier within all transition zones. Some pre- or postzygotic isolation mechanisms (habitat specialization, spawning asynchrony, assortative fertilization and hybrid depression) have been documented in previous studies, although their relative importance remains to be evaluated. We also provided evidence for a recent migratory 'short-cut' connecting M. edulis-like populations of the Charente patch to an external M. edulis population in Normandy and thought to reflect artificial transfer of spat for aquaculture. PMID- 12535096 TI - Glacial refugia, haplotype distributions, and clonal richness of the Daphnia pulex complex in arctic Canada. AB - As part of a large international Arctic biodiversity expedition (Tundra Northwest '99), we examined the distribution of members of the arctic Daphnia pulex complex (Cladocera, Anomopoda) from 121 tundra ponds, spread across 16 sites spanning a large portion of arctic Canada (i.e. from 62 degrees 22' N to 79 degrees 01' N; 66 degrees 45' W to 139 degrees 37' W). Using allozyme electrophoresis and mitochondrial (mt)DNA analyses, we examined the population genetic (clonal) structure of these populations. The following taxa were detected in this complex: Daphnia pulicaria, D. middendorffiana and D. tenebrosa. Clear geographical differences in mean clonal richness and diversity were observed, with most western sites exhibiting higher clonal richness and diversity, than sites in the eastern Canadian Arctic. For both the pulicaria group (i.e. D. pulicaria and D. middendorffiana) and D. tenebrosa, the highest mean regional clonal richness was detected from the southern section of Banks Island, an unglaciated site situated on the edge or directly in the eastern fringe of the Beringian glacial refuge. A significant negative correlation was found between geographical distance from the Beringian edge, and overall regional clonal richness (i.e. sites closer to the edge harboured greater clonal richness). These results clearly indicate that more recently deglaciated regions (i.e. eastern Canadian Arctic) harbour lower levels of clonal richness than western regions nearer Beringia. We discuss the role that glacial refugia have played in influencing both biotic and genetic diversity in arctic taxa. PMID- 12535097 TI - Paternity success and the direction of sexual selection in a field population of a semelparous marsupial, Antechinus agilis. AB - Antechinus agilis is a small sexually size dimorphic marsupial with a brief annual mating period of 2-3 weeks. All males die after this period, and females give birth to up to 10 young. Mating is thought to be promiscuous, however, there is no field data to confirm this. Using microsatellites, we investigated paternity patterns over two seasons in a wild population. Male weight was significantly positively related to the number of females fertilized and with the number of offspring sired, in both years. Furthermore, selection gradients indicated selection for larger males. Both results suggest that size dimorphism in A. agilis can be explained by sexual selection for larger males. The proportion of offspring sired within litters, did not relate to male size. Therefore, larger males are more successful through higher mating access, not through their sperm outcompeting that of smaller males. As expected from their known ranging behaviour, the number of offspring within litters left unassigned to a father did not depend on the grid location of the mother. Female size did not differ between successful reproducing and unsuccessful females. However, females that weaned offspring had larger heads than females that did not wean offspring. Males did not 'prefer' mating with larger females, nor did assortative mating occur. From our results, the mating system of A. agilis is clearly promiscuous. Selection for larger males occurred in both years, even though in one year the operational sex ratio was highly female biased, suggesting that the potential reproductive rate is a better predictor of the direction of sexual selection in A. agilis. PMID- 12535098 TI - Sperm length influences fertilization success during sperm competition in the snail Viviparus ater. AB - Sperm form and size is tremendously variable within and across species. However, a general explanation for this variation is lacking. It has been suggested that sperm size may influence sperm competition, and there is evidence for this in some taxa but not others. In addition to normal fertilizing sperm, a number of molluscs and insects produce nonfertile sperm that are also extremely morphologically variable, and distinct from fertilizing forms. There is evidence that nonfertile sperm play an indirect role in sperm competition by decreasing female remating propensity in Lepidopterans, but in most taxa the function of parasperm is unknown. We investigated the role of nonfertile (oligopyrene) sperm during sperm competition in the fresh water snail Viviparus ater. Previous studies found that the proportion of oligopyrene sperm increased with the risk of sperm competition, and hence it seems likely that these sperm influence fertilization success during competitive matings. In mating experiments in which females were sequentially housed with males, we examined a range of male characteristics which potentially influence fertilization success. We found that the size of oligopyrene sperm was the best predictor of fertilization success, with males having the longer sperm siring the highest proportion of offspring. Furthermore, we found a positive shell size and sperm concentration effect on paternity, and females with multiply sired families produced more offspring than females mating with only one male. This result suggests polyandry is beneficial for female snails. PMID- 12535099 TI - Impact of oilseed rape expressing the insecticidal cysteine protease inhibitor oryzacystatin on the beneficial predator Harmonia axyridis (multicoloured Asian ladybeetle). AB - Insect-resistant transgenic plants have been suggested to have deleterious effects on beneficial predators through transmission of the transgene product by the pest to the predator. To test this hypothesis, effects of oilseed rape expressing the cysteine protease inhibitor oryzacystatin-1 (OC-1) on the predatory ladybird Harmonia axyridis were investigated using diamondback moth Plutella xylostella as the pest species. As expected, oilseed rape expressing OC 1 had no effects on either development or survival of the pest, which utilizes serine digestive proteases. Immunoassays confirmed accumulation of the transgene product in pest larval tissues at levels of up to 3 ng per gut. Characterization of proteolytic digestive enzymes of H. axyridis demonstrated that larvae and adults utilize cysteine and aspartic proteases; the former activity was completely inhibited by oryzacystatin in vitro. However, when H. axyridis larvae consumed prey reared on OC-1 expressing plants over their entire life cycle, no significant effects upon survival or overall development were observed. The inhibitor initially stimulated development, with a shortening of the developmental period of the second instar by 27% (P < 0.0001) accompanied by a 36% increase in weight of second instar larvae (P = 0.007). OC-1 had no detrimental effects on reproductive fitness of adult H. axyridis. Interestingly there was a significant increase in consumption of OC-1 dosed prey. The results show that prey reared on transgenic plants expressing a protein which inhibited ladybird digestive enzymes in vitro had no effects in vivo; the ladybird was able to up-regulate digestive proteases in response to the inhibitor. PMID- 12535100 TI - Habitat differentiation vs. isolation-by-distance: the genetic population structure of Elymus athericus in European salt marshes. AB - We investigated genetic differentiation among populations of the clonal grass Elymus athericus, a common salt-marsh species occurring along the Wadden Sea coast of Europe. While E. athericus traditionally occurs in the high salt marsh, it recently also invaded lower parts of the marsh. In one of the first analyses of the genetic population structure in salt-marsh species, we were interested in population differentiation through isolation-by-distance, and among strongly divergent habitats (low and high marsh) in this wind- and water-dispersed species. High and low marsh habitats were sampled at six sites throughout the Wadden Sea. Based on reciprocal transplantation experiments conducted earlier revealing lower survival of foreign genotypes we predicted reduced gene flow among habitats. Accordingly, an analysis with polymorphic cross-species microsatellite primers revealed significant genetic differentiation between high and low marsh habitats already on a very small scale (< 100 m), while isolation by-distance was present only on larger scales (60-443 km). In an analysis of molecular variance we found that 14% of the genetic variance could be explained by the differentiation between habitats, as compared to only 8.9% to geographical (isolation-by-distance) effects among six sites 2.5-443 km distant from each other. This suggests that markedly different selection regimes between these habitats, in particular intraspecific competition and herbivory, result in habitat adaptation and restricted gene flow over distances as small as 80 m. Hence, the genetic population structure of plant species can only be understood when considering geographical and selection-mediated restrictions to gene flow simultaneously. PMID- 12535101 TI - Variation of haplotype distributions of two genomic regions of Citrus tristeza virus populations from eastern Spain. AB - Genetic variation in natural populations of Citrus tristeza virus (CTV) was studied using haplotypes detected by single-strand conformation polymorphism (SSCP) analysis of two genomic regions (p20 gene and segment A, located in ORF1a). Analysis of 254 samples from 125 trees, collected at 12 different sites, yielded 8 different haplotypes for p20 and 5 for segment A. The most frequent haplotype of p20 was predominant at all sites, but several sites differed in the predominance of segment A haplotypes. At most sites, the homozygosity observed for the p20 gene tended to be higher than expected in a neutral evolution, whereas the opposite was true for segment A. Comparison of the populations at different sites showed that 44 of the 66 possible population pairs were genetically distinct for segment A, but only six pairs differed for the p20 gene. Analysis of molecular variance grouping trees by site, scion variety, rootstock or age, showed that variation in segment A was significantly affected by site, tree age and rootstock, and that variation between trees in each group and within trees was even more important. In contrast, variation in p20 was affected only by site and rootstock, each factor contributing to < 2% of the variation. The data suggest that sequence variations in segment A must be functionally less important and that it has less evolutionary constraints than p20. Detection of different haplotypes in neighbour trees or in samples from the same tree may help explain part of the variability observed in CTV symptom expression. PMID- 12535102 TI - The clonal structure of Quercus geminata revealed by conserved microsatellite loci. AB - The scrub oak communities of the southeastern USA may have existed at their present locations for thousands of years. These oaks form suckers, and excavations of root systems suggest that clones may occupy very large areas. Resolution of the clonal nature of scrub oaks is important both to manage the tracts of this ecosystem that remain, and in conducting long-term ecological studies, where the study area must substantially exceed the area occupied by any single clone. Microsatellites were used to determine the genetic diversity of a dominant oak species within a 2-ha long-term experimental site on Merritt Island at the Kennedy Space Center. This area contains a long-term study of the effects of elevated CO2 on the ecosystem. Conservation of seven microsatellite loci, previously identified in the sessile oak, Quercus petraea, was tested in two Florida scrub oak species, Q. geminata and Q. myrtifolia. Sequence analysis revealed that all seven microsatellite loci were conserved in Q. geminata and five loci were conserved in Q. myrtifolia. Six microsatellite loci were polymorphic in Q. geminata and these were subsequently used to investigate the clonal structure of the Q. geminata population. Twenty-one unique combinations of microsatellites, or haplotypes, occurred only once, whereas the remaining 26 individuals belonged to a total of seven different haplotypes. Trees with identical haplotypes were in close proximity, supporting the interpretation that they were clones. The results showed that there is significant genetic diversity within the 2-ha experimental site. Microsatellites provided a powerful and noninvasive tool for distinguishing individual genotypes and determining an adequate area for long-term ecosystem studies. PMID- 12535103 TI - Confirmation of low genetic diversity and multiple breeding females in a social group of Eurasian badgers from microsatellite and field data. AB - The Eurasian badger (Meles meles) is a facultatively social carnivore that shows only rudimentary co-operative behaviour and a poorly defined social hierarchy. Behavioural evidence and limited genetic data have suggested that more than one female may breed in a social group. We combine pregnancy detection by ultrasound and microsatellite locus scores from a well-studied badger population from Wytham Woods, Oxfordshire, UK, to demonstrate that multiple females reproduce within a social group. We found that at least three of seven potential mothers reproduced in a group that contained 11 reproductive age females and nine offspring. Twelve primers showed variability across the species range and only five of these were variable in Wytham. The microsatellites showed a reduced repeat number, a significantly higher number of nonperfect repeats, and moderate heterozygosity levels in Wytham. The high frequency of imperfect repeats and demographic phenomena might be responsible for the reduced levels of variability observed in the badger. PMID- 12535104 TI - Widespread occurrence of a domestic dog mitochondrial DNA haplotype in southeastern US coyotes. AB - Sequence analysis of the mitochondrial DNA control region from 112 southeastern US coyotes (Canis latrans) revealed 12 individuals with a haplotype closely related to those in domestic dogs. Phylogenetic analyses grouped this new haplotype in the dog/grey wolf (Canis familiaris/Canis lupus) clade with 98% bootstrap support. These results demonstrate that a male coyote hybridized with a female dog, and female hybrid offspring successfully integrated into the coyote population. The widespread distribution of this haplotype from Florida to West Virginia suggests that the hybridization event occurred long ago before the southeastern USA was colonized by coyotes. However, it could have occurred in the southeastern USA before the main front of coyotes arrived in the area between male coyotes released for sport and a local domestic dog. The introgression of domestic dog genes into the southeastern coyote population does not appear to have substantially affected the coyote's genetic, morphological, or behavioural integrity. However, our results suggest that, contrary to previous reports, hybridization can occur between domestic and wild canids, even when the latter is relatively abundant. Therefore, hybridization may be a greater threat to the persistence of wild canid populations than previously thought. PMID- 12535106 TI - Nuclear DNA identification of migrating bull trout captured at the Puget Sound Energy diversion dam on the White River, Washington State. AB - Bull trout (Salvelinus confluentus) is a char listed as threatened under the United States Endangered Species Act throughout its range in the coterminous United States. Substantial morphological similarities between bull trout and Dolly Varden (S. malma) make field identification difficult. This has resulted in an incomplete understanding of their distribution and abundance in Washington State where these two species occur sympatrically. We used three diagnostic nuclear loci to determine the species of char collected at a trap on the White River in southern Puget Sound (Washington State, USA). Each of the 104 samples revealed the expected bull trout genotype at all three loci. This work presents three principle results: (i) the presence of a migratory bull trout population in southern Puget Sound; (ii) no evidence of migratory Dolly Varden over 3 years; and (iii) no evidence of hybridization was detected. These results also demonstrate how molecular markers can provide information essential to the conservation and management of these species. PMID- 12535105 TI - Phylogeography and molecular rates of subterranean aquatic Stenasellid Isopods with a peri-Tyrrhenian distribution. AB - The subterranean Isopods belonging to the genus Stenasellus have an interesting disjunct distribution in the peri-Tyrrhenian area with morphologically closely related taxa occurring in Sardinia, Corsica, Tuscany and in the Pyrenees phreatic and interstitial waters. Because the dispersal capacities of these organisms are limited, their distribution has been associated traditionally with the tectonic events leading to the separation of the Sardinia-Corsica microplate from the Pyrenees and its subsequent movement towards the Italian peninsula. We sequenced a fragment of the mtDNA cytochrome oxidase I gene (COI) for multiple populations of the S. racovitzai species-group (Corsica, Sardinia, Tuscany) and S. virei (Pyrenees). We found that multiple phylogenetic analyses always gave the same topology, which is consistent with the genetic relations found using allozyme data, and with the palaeogeography of the area. The molecular data suggest that a combination of vicariance and dispersal events explain most effectively the present distribution pattern of these organisms. We also calculated COI rates and calibrated them against absolute time, taking advantage of the availability of two geologically based time estimates. Rates on all substitutions are similar to those published for other crustaceans for the same COI fragment, including taxonomically and ecologically distant groups. Rates on third codon positions or on transversions are generally lower than those found in other crustaceans. PMID- 12535108 TI - New century, new challenges for community based medical education. PMID- 12535109 TI - Rural medical education: how different is it? PMID- 12535110 TI - Techniques used by "expert" and "non-expert" tutors to facilitate problem-based learning tutorials in an undergraduate medical curriculum. AB - BACKGROUND: There is inconclusive debate within the literature as to whether the best problem-based learning (PBL) tutors are subject experts or not. The debate hinges on whether knowledgeable tutors are tempted to intervene too often in PBL discussions compared to non-expert tutors, and whether the latter may not be able to sufficiently challenge the students' level of understanding. PURPOSE: To describe approaches used by tutors in PBL tutorials and to identify differences between tutors from medical and non-medical backgrounds. METHODS: The research reported in this paper was undertaken during the academic session 1999-2000 at the University of Liverpool Faculty of Medicine. A qualitative exploratory case study method was used and two PBL groups were observed. One of these groups had a medically qualified tutor and the other had a tutor from a humanities background. The focus of the observation was the discourse between tutor and students, which was analysed using a framework drawn from linguistics. Results were fed back to both the tutors and the students to check their perceptions of the interactions. RESULTS: Analysis of the tutorial group interaction revealed that tutors from both backgrounds used similar techniques to raise students' awareness, facilitate the group process and direct students' learning. Differences were noted between the two tutors: the medical tutor set out to raise students' awareness by using questioning techniques herself, whereas the non-medical tutor expected students to question each other. The non-medical tutor was observed to facilitate the group process more often than the medical tutor. CONCLUSIONS: Qualitative analysis of spoken discourse in PBL tutorials provides valuable insights into the processes involved in PBL, thereby generating material which is useful for both training of and giving feedback to PBL tutors. PMID- 12535111 TI - Does problem-based learning lead to deficiencies in basic science knowledge? An empirical case on anatomy. AB - INTRODUCTION: Problem-based learning (PBL) is supposed to enhance the integration of basic and clinical sciences. In a non-integrative curriculum, these disciplines are generally taught in separate courses. Problem-based learning students perceive deficiencies in their knowledge of basic sciences, particularly in important areas such as anatomy. Outcome studies on PBL show controversial results, sometimes indicating that medical students at PBL schools have less knowledge of basic sciences than do their colleagues at more traditional medical schools. We aimed to identify differences between PBL and non-PBL students in perceived and actual levels of knowledge of anatomy. METHODS: Samples of Year 4 students in all eight medical schools in the Netherlands completed a questionnaire on perceived knowledge and took part in a computerised anatomy test consisting of both clinically contextualised items and items without context. RESULTS: Problem-based learning students were found to have the same perceived level of anatomy knowledge as students at other medical schools. Differences in actual levels of knowledge were found between schools. No significant effects on knowledge levels were found for PBL schools versus non-PBL schools. CONCLUSION: The results of this study show that PBL does not result in a lower level of anatomy knowledge than more traditional educational approaches. It remains to be ascertained whether the levels students attain are adequate. Subjects for further study are the desired level of anatomy knowledge at the end of undergraduate medical education and the effectiveness of basic science learning within a clinical context and with repetition over the course of the curriculum. PMID- 12535112 TI - Hitting the mark: negotiated marking and performance factors in the communication skills element of the VOICE examination. AB - INTRODUCTION: Communication skills assessment is complex. Standardised patient use is widespread, but anxiety exists around the use of role players as assessors of competence in high stakes examinations. This study measures the level of agreement between scoring examiners and role players, and considers their influence on each other. Examiner status and question choices are analysed as variables. METHOD: The valid oral interactive contextualised examination (VOICE) is a general practice examination styled as an objective structured clinical examination (OSCE) of six 15-minute stations, which include two role-played consultations with professional role players. The examination candidates are final year medical students. Clinical components are examined by a general practitioner (GP). Communication skills are assessed by these examiners in conjunction with the role players, through a process of negotiation. Descriptive professionalism/attitude bandings are used as percentage-scoring guidelines. Checklists are not used. For this study, the initial (independently) perceived marks of the two scoring groups and their agreed final (awarded) marks were recorded, along with other variables including gender, performance factors, demographics and the nature of the question. Data represents 512 students undertaking 1024 simulated consultations, examined by 28 role players and 46 examiners. Analysis was carried out using SPSS Version 10. RESULTS: Results show that the examination and negotiation process is consistent. Role players have a direct influence on scoring. The examiner's background is a significant variable [F9,1014 = 4.207, P < 0.001]. Students perform less well on questions involving higher degrees of clinical information giving. Question choice is not significant [F30,3039 = 1.397, P=0.074]. DISCUSSION: The variables in the examination do not indicate any discrepancy substantial enough to bias a student's grade. Negotiated marking in this context is considered safe and reliable. PMID- 12535113 TI - Perceived stress during undergraduate medical training: a qualitative study. AB - INTRODUCTION: Medical education is long and emotionally taxing. It can involve levels of stress that lead to disruptions in both physical and mental health. This qualitative study explores the views of Year 5 medical students on the causes of stress throughout their undergraduate medical training. METHOD: Semi structured interviews were conducted with 21 final year medical students at the University of Birmingham between January and May 2001. RESULTS: Pressure of work, especially in terms of preparing for examinations and acquiring professional knowledge, skills and attitudes were reported as the most stressful aspects of medical training. Transition periods, particularly between school and medical school, preclinical and clinical training, and clinical training to approaching qualification were highlighted as particularly stressful. A perceived lack of support from the medical school authorities also appeared to add to student stress levels. DISCUSSION: Student stress may be alleviated by greater guidance and support from the medical school during crucial transition periods. Aspects of professional socialisation may also need to be addressed to reduce the levels of stress associated with undergraduate training for future generations of medical students. PMID- 12535114 TI - Sexual and gender-related harassment in medical education and research training: results from a Swedish survey. AB - OBJECTIVE: The aims of this study were to establish the level of perceived sexual and gender-related harassment in undergraduate and doctoral studies, in which environment the events occurred, which categories of persons had committed the harassment, and other aspects of sexual harassment at the Faculty of Medicine, Gothenburg University. METHODS: A questionnaire was distributed to all registered male and female undergraduate students (n= 605) and doctoral students (n=743) by mail to their home addresses. RESULTS: The response rate was 62% (840/1348). Of the total study population, 59% (495/840) of respondents reported at least one experience of derogatory jokes and comments, 54% (454/840) of respondents reported at least one experience of gender-related discrimination, and 22% (187/840) of respondents reported at least one incident of sexual harassment. More severe types of sexual harassment were reported by 9% (79/840) of respondents. Women, and especially undergraduate women, were more often exposed to all kinds of harassment than were men. Lecturers/professors, doctors and co students were the categories most often identified as the harassers. The harassment mostly occurred during lectures, clinical work and coffee breaks. The most common types of self-perceived mistreatment were derogatory jokes and comments. CONCLUSION: This survey shows that sexual harassment happens to both men and women, although it is more commonly experienced by female undergraduate and doctoral students, and that it occurs in both the university and hospital environments. Universities should develop action plans to prevent such events. Students and teachers should be well informed about appropriate measures to take in situations where harassment is known or suspected to occur. PMID- 12535115 TI - Early clinical exposure to people who are dying: learning to care at the end of life. AB - BACKGROUND: The nature of medical care at the end of life and, in particular, the way in which caring is learned remain problematic for medical educators and the profession. Recent work has indicated that doctors learn to care, in an emotional and intimate way, from people who are dying. METHODS: This paper reports on the development of a programme designed for medical students in their first clinical year who spend time with a person who is dying and their family. The students are required to produce a portfolio assignment that includes a personal reflection of the experience. The findings from a phenomenological study undertaken using these personal reflections are reported. These reflections and comments are interpreted as being embedded in five key themes. RESULTS: The actual encounters differed from the medical students' anticipation of them. Students identified an emotional component to the experience; they explored their own and the patient's understandings of spirituality; they reflected on personal meanings of the encounter and they suggested ways in which they might learn to care more effectively for people who are dying. DISCUSSION: The way in which many of these students approach end-of-life care has been altered through a transformative educational experience that encouraged them to draw on their own experiences and skills. Their learning was facilitated by the writing of accounts and the discussion that each group held with teaching staff at the conclusion of the programme. PMID- 12535116 TI - Learning from lives: a model for health and social care education in the wider community context. AB - OBJECTIVES: To develop and evaluate a model for medical education which draws upon the whole frontline workforce in primary care and which enables medical students to develop skills and competencies in patient-centred teamworking across organisational boundaries. METHODS: Over a period of 3 years, 517 undergraduate medical students undertook a 4-week community hospital-based attachment developed in partnership with frontline staff. Pre- and post-course questionnaires and qualitative evaluation were sought from students, patients, tutors and frontline staff. RESULTS: The performance of students in assessment was very good. Before receiving the assessment results, students perceived a high degree of achievement of the specified learning outcomes and reported significant changes in attitude. Qualitative comments were overwhelmingly positive, with clear indications that students appreciated the unique learning opportunities available in this model. Patients were very positive about continuing involvement and valued the opportunity to influence future doctors. Staff were enthusiastic and committed despite competing service pressures. CONCLUSIONS: We have successfully developed a partnership model of community-based education and shown that it leads to significant changes in attitude in students and enables them to learn in an active, patient-focused way about the complexities of delivery of care outside the secondary care environment. We have identified the key requirements for implementation of the model in other centres. PMID- 12535117 TI - Evaluating the spoken English proficiency of international medical graduates: detecting threats to the validity of standardised patient ratings. AB - PURPOSE: To investigate potential threats to the validity of the spoken English proficiency ratings provided by standardised patients (SPs) in high-stakes clinical skills examinations. METHOD: Spoken English ratings from 43 327 patient encounters were studied. These involved over 5000 candidates, 40% of whom were female and 33% of whom self-reported English to be their native language. Over 100 SPs were involved in the study, 51% of whom were female and 90% of whom were native English speakers. Possible performance differences in English ratings were studied as a function of candidate and SP gender, and as a function of candidate and SP native language (English versus all other languages). RESULTS: No significant candidate by SP gender effect was detected. There were no meaningful differences in mean English ratings as a function of SP or candidate gender. Likewise, English ratings did not vary as a function of either candidate or SP native language. While candidate mean English ratings were not associated with the native language of the SP, native English-speaking candidates did achieve significantly higher ratings. DISCUSSION: The lack of significant interaction between candidate and SP gender, and candidate and SP native language, suggests that the SPs provided unbiased English ratings. These results, combined with the expected higher English ratings given to candidates with English-speaking backgrounds, provides additional evidence to support the validity and fairness of spoken English proficiency ratings provided by standardised patients. PMID- 12535118 TI - Bottom drawer papers. PMID- 12535119 TI - Overseas-trained refugee doctors. PMID- 12535120 TI - Computer-aided surgical training. PMID- 12535121 TI - Lambert Schuwirth. PMID- 12535123 TI - Rapid reduction of Legionella pneumophila on stainless steel with zeolite coatings containing silver and zinc ions. AB - AIMS: To determine the rate of reduction of Legionella pneumophila by stainless steel surfaces with zeolite ceramic coatings containing 2.5% (w/w) silver (Ag) and 14% zinc (Zn) ions. METHODS AND RESULTS: Stainless steel pans with and without Ag/Zn coatings were inoculated with solutions of Leg. pneumophila ATCC 33155 and incubated at 37 degrees C. Survival was monitored using the spread plate technique on selective buffered charcoal yeast extract agar. Significant reductions of Leg. pneumophila were effected by the Ag/Zn zeolite coatings within 2 h of exposure. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: Zeolite ceramic Ag/Zn coatings impart significant anti-Legionella properties to stainless steel surfaces. Coated stainless steel could be used in the manufacture of air ducts, condensation pans and intake and exhaust vents. These products have the potential to reduce numbers of Legionella in air-handling systems. PMID- 12535124 TI - Fate of Escherichia coli O157:H7 during the manufacture of Mozzarella cheese. AB - AIMS: The fate of Escherichia coli O157:H7 was investigated during the manufacture of Mozzarella cheese. METHODS AND RESULTS: The Mozzarella cheese was made from unpasteurized milk which was inoculated to contain ca 10(5) cfu ml( 1)E. coli O157:H7. Two different heating temperatures (70 and 80 degrees C), commonly used during curd stretching, were investigated to determine their effects on the viability of E. coli O157:H7 in Mozzarella cheese. Stretching at 80 degrees C for 5 min resulted in the loss of culturability of E. coli O157:H7 strains, whereas stretching at 70 degrees C reduced the number of culturable E. coli O157:H7 by a factor of 10. CONCLUSIONS: The results show that stretching curd at 80 degrees C for 5 min is effective in controlling E. coli O157:H7 during the production of Mozzarella cheese. Brining and storage at 4 degrees C for 12 h was less effective than the stretching. SIGNIFICANCE AND IMPACT OF THE STUDY: Mozzarella cheese should be free of E. coli O157:H7 only if temperatures higher than or equal to 80 degrees C are used during milk processing. PMID- 12535126 TI - Control of pathogenic Vibrio spp. by Bacillus subtilis BT23, a possible probiotic treatment for black tiger shrimp Penaeus monodon. AB - AIMS: The present study evaluated the in vitro and in vivo antagonistic effect of Bacillus against the pathogenic vibrios. METHODS AND RESULTS: Cell-free extracts of Bacillus subtilis BT23 showed greater inhibitory effects against the growth of Vibrio harveyi isolated by agar antagonism assay from Penaeus monodon with black gill disease. The probiotic effect of Bacillus was tested by exposing shrimp to B. subtilis BT23 at a density of 106-108 cfu ml-1 for 6 d before a challenge with V. harveyi at 103-104 cfu ml-1 for 1 h infection. The combined results of long- and short-term probiotic treatment of B. subtilis BT23 showed a 90% reduction in accumulated mortality. CONCLUSIONS: This study reports that pathogenic vibrios were controlled by Bacillus under in vitro and in vivo conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Results indicated that probiotic treatment offers a promising alternative to the use of antibiotics in shrimp aquaculture. PMID- 12535125 TI - Characterization of amplified polymerase chain reaction glnB and nifH gene fragments of nitrogen-fixing Burkholderia species. AB - AIMS: To clone and sequence polymerase chain reaction (PCR)-amplified glnB and nifH genes of the nitrogen-fixing bacteria Burkholderia brasilensis strain M130, B. tropicalis strain PPe8 and B. kururiensis strain KP23. METHODS AND RESULTS: The glnB and nifH gene fragments were amplified by PCR using universal degenerated primers. A very high percentage of similarity for the nifH (100%) and glnB (96%) genes was observed between strains M130 and KP23. A similarity of 100% for the nifH gene was also observed between strains M130 and PPe8. However, the identity for the glnB gene was 98% and the similarity 88%. The phylogenetic tree of the nifH gene showed a very high degree of similarity to the 16S rDNA gene. CONCLUSIONS: The nitrogen-fixing bacteria of the Burkholderia genus formed a cluster separated from the other species of the genus mainly when the nifH rather than the glnB gene was used to construct the phylogenetic tree. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of the nifH and glnB gene sequences of B. brasilensis, B. tropicalis and B. kururiensis will support new studies on the diversity of these diazotrophs in natural environments. PMID- 12535127 TI - Identification of Thiothrix unzii in two distinct ecosystems. AB - AIMS: Molecular procedures were used to identify Thiothrix spp. in biofilms from sulphide-rich waters in two distinct ecosystems. METHODS AND RESULTS: Biofilm samples were obtained from two groundwater-fed systems in central and northern Florida, including an artesian spring and municipal water tank. The 16S rDNA in each sample was directly amplified by polymerase chain reaction. CONCLUSIONS: Clonal libraries of biofilm 16S rDNA from each site contained rDNA sequences that were 99-99.5% similar to Thiothrix unzii. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of T. unzii in a natural system. Biofilm formation by Thiothrix spp. can cause fouling in groundwater processing equipment, including municipal water-processing facilities, agricultural irrigation systems and spring water bottling plant filters. Biofouling can have severe economic and human health impacts as it will influence flow rates and related water treatments. Characterization of specific fouling bacteria and their molecular ecology is essential for their regulation. PMID- 12535128 TI - Levels of male-specific RNA bacteriophage and Escherichia coli in molluscan bivalve shellfish from commercial harvesting areas. AB - AIMS: Current measures for controlling the public health risks associated with bivalve molluscan shellfish consumption rely on the use of Escherichia coli to indicate the sanitary quality of shellfish harvesting areas. However, it has been demonstrated that E. coli is an inadequate indicator of the viral risk associated with shellfish. An alternative indicator organism, male-specific RNA (FRNA) bacteriophage has been proposed for this role. This study compared the distribution of E. coli and FRNA bacteriophage in shellfish harvesting areas. METHODS AND RESULTS: A total of 608 shellfish samples from 49 shellfish harvesting areas were analysed for E. coli and FRNA bacteriophage using standard published methods. The geometric mean concentration of FRNA bacteriophage in all samples was over three times greater than that of E. coli (1800 and 538 counts/100 g for FRNA bacteriophage and E. coli, respectively). In contrast to E. coli, FRNA bacteriophage concentrations were strongly influenced by season with a geometric mean count of 4503 PFU/100 g in the winter (October-March) compared with 910 PFU/100 g in the summer (April-September). CONCLUSIONS: FRNA bacteriophage were present in shellfish at higher concentrations than E. coli. Elevated levels of FRNA bacteriophage observed in the winter concur with the known increased viral risk associated with shellfish harvested at that time of year in the UK. Levels of FRNA bacteriophage found in many shellfish from category B harvesting areas would not be eliminated by conventional treatment processes. SIGNIFICANCE AND IMPACT OF THE STUDY: Data from this study will inform future proposals to introduce FRNA bacteriophage as an indicator of the viral risk associated with shellfish. PMID- 12535129 TI - Microemulsions are highly effective anti-biofilm agents. AB - AIMS: The demonstration of the antibiofilm effects of pharmaceutical microemulsions. METHODS AND RESULTS: Microemulsions were prepared as physically stable oil/water systems. Previous work by this group has shown that microemulsions are highly effective antimembrane agents that result in rapid losses of viability in planktonic populations of Pseudomonas aeruginosa and Staphylococcus aureus. In this experiment a microemulsion preparation was used upon established biofilm cultures of Ps. aeruginosa PA01 for a period of 4 h. The planktonic MIC of sodium pyrithione and the planktonic and biofilm MICs of cetrimide were used as positive controls and a biofilm was exposed to a volume of normal sterile saline as a treatment (negative) control. Results indicate three log-cycle reductions in viability within the microemulsion treated biofilm, as compared to those observed in control treatments of similar biofilms (one log cycle reduction in viabilities). CONCLUSIONS: The results indicate that the microemulsions are highly effective antibiofilm agents. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that microemulsions may have a role in the treatment of industrial and environmental biofilms. PMID- 12535130 TI - Comparison of sample collection methods for the PCR detection of oral anaerobic pathogens. AB - AIMS: To provide evidence that DNA-PCR diagnostics of oral pathogens based on standard sample collection by paper point insertion from the depth of the periodontal pocket can be replaced by a novel non-invasive collection method based on swab technique from the gingiva. METHODS AND RESULTS: In this study we compared the results from two collection methods performed in 35 patients with chronic adult periodontitis. Statistical analysis showed a highly significant association of diagnostic results between both collection techniques. CONCLUSIONS: The Pocket-out method represents a reliable alternative to the standard collection technique for PCR diagnosis of oral pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Due to its simplicity and non-invasiveness, the Pocket out collection could be performed in any physician office, or even by the patient himself. With respect to the putative association between periodontal disease and various systemic illnesses, this method could be integrated with various screening programs of oral pathogens. PMID- 12535131 TI - Reference isolates for the clonal complexes of Campylobacter jejuni. AB - AIMS: To identify and make available through the National Collection of Type Cultures (NCTC) a set of reference isolates for the clonal complexes of Campylobacter jejuni. METHODS AND RESULTS: The development of a multilocus sequence typing scheme for C. jejuni enabled the genetic characterization of a large number of isolates (n = 814) from cases of human disease, animals, birds and their food products. The nucleotide sequence data were used to assign each isolate an allelic profile or sequence type (ST) and examine the C. jejuni population structure in terms of clonal complexes. The clonal complexes consisted of an abundant central or founder genotype (ST), after which the complex was named, together with very closely related, generally less abundant genotypes differing from the founder at one, two or three loci. The clonal complex is an informative unit for the study C. jejuni epidemiology. It provides data which enabled the choice of 13 C. jejuni founder isolates for submission to the NCTC as a representative cross-section of the C. jejuni population. CONCLUSIONS: These 13 isolates provide a defined resource for further research into aspects of C. jejuni biology such as genomic diversity, virulence and adaptation to particular hosts or environmental survival. SIGNIFICANCE AND IMPACT OF STUDY: This isolate collection is available through the NCTC and provides a resource for further research. PMID- 12535132 TI - Antifungal activity of the essential oil of Agastache rugosa Kuntze and its synergism with ketoconazole. AB - AIMS: To evaluate the fungitoxic activity of the essential oil of Agastache rugosa alone and to determine its combination effect with ketoconazole against Blastoschizomyces capitatus. METHODS AND RESULTS: The antifungal activities of the essential oil of A. rugosa and its main constituent estragole were investigated using the broth microdilution, disk diffusion methods and checkerboard microtitre assay. Both estragole and the essential oil exhibited strong activities against the tested fungi and showed synergism with ketoconazole against B. capitatus. CONCLUSIONS: Both estragole and the essential oil of A. rugosa have significant growth-inhibiting activities against B. capitatus showing strong synergistic effect with ketoconazole. SIGNIFICANCE AND IMPACT OF THE STUDY: The essential oil of A. rugosa, combined with ketoconazole, may be particularly useful against B. capitatus, a rare pathogenic fungus documented to cause severe and fatal mycoses in immunocompromised patients. PMID- 12535133 TI - Use of polymerase chain reaction to detect the soft rot pathogen, Pythium myriotylum, in infected ginger rhizomes. AB - AIMS: The aims are to establish a polymerase chain reaction (PCR)-based method for detecting Pythium myriotylum in the rhizome of ginger and diagnosing ginger soft rot and screening health seed ginger. METHODS AND RESULTS: A booster PCR method was established for detection of P. myriotylum using a specific primer selected from rDNA ITS1 region coupled with universal primer ITS2. It successfully applied to the detection of P. myriotylum in naturally infected ginger rhizomes but not from DNA of ginger rhizomes collected from field without target fungus. CONCLUSIONS: A specific method for detecting P. myriotylum was achieved. SIGNIFICANCE AND IMPACT OF THE STUDY: The new PCR method has allowed us to monitor ginger for the presence of P. myriotylum as a way of disease diagnosis or healthy seed ginger examination. PMID- 12535134 TI - Expression of an artificial polypeptide with a repeated tripeptide glutamyl tryptophanyl-lysine in Saccharomyces cerevisiae. AB - AIMS: Artificial genes, which encode 48 or 64 repeats of a tripeptide, glutamyl tryptophanyl-lysine have been cloned to the yeast expression vector pAM82 containing the PHO5 promoter and expressed in Saccharomyces cerevisiae AH22. METHODS AND RESULTS: When the yeast cells harbouring recombinant plasmids pALTG6 2 and pALTG4-4 were derepressed in Burkholder minimal medium (Toh-e, A., Ueda, Y., Kakimoto, S.I. and Oshima, Y. (1973) Journal of Bacteriology113, 727-738) containing low phosphate (0.03 g l-1 KH2PO4 and 1.5 g l-1 KCl), the expression was the highest after 24 h induction and the artificial polypeptides were synthesized to about 10% (pALTG6-2) and 14% (pALTG4-4) of the total cell protein. CONCLUSIONS: The artificial polypeptides produced in yeast were made to react with the rabbit antiserum against the polypeptide purified from Escherichia coli and found only in the pellet fraction of cell lysates, indicating the formation of inclusion body. Artificial polypeptide consisting of Glu-Trp-Lys may be useful as partial supplement in food and feeds. SIGNIFICANCE AND IMPACT OF THE STUDY: The production of single cell enriched with homopolymers of an essential amino acid in yeast might be an important tool of supplementing cereal diets and feed grain rations and could be used as means for improvement of the amino acid profile of single cell protein and production of pharmaceutical peptides. PMID- 12535135 TI - Reliability of clinician judgements of bruxism. AB - Bruxism is considered to be a parafunctional disorder requiring treatment and is viewed as a risk factor for the development of temporomandibular disorders (TMDs). The purpose of this investigation is to examine the reliability of clinician judgements of bruxism severity. Twenty dentists who are faculty members in a dental school examined 29 stone casts and gold-plated models of individual teeth for evidence of bruxism. Ordinal ratings of bruxism severity for the 29 augmented models were made on two occasions, approximately 3 months apart. Inter rater reliability among all clinicians, evaluated using intraclass correlation coefficients (ICCs), was poor at both time one and time two (i.e. ICC = 0.33 and 0.32, respectively), with somewhat better reliability found among those clinicians with above-average time elapsed since completion of dental training (i.e. ICC = 0.48 and 0.50 for time 1 and time 2, respectively). Three-month test retest reliabilities were fair (ICC = 0.46) for the full group of raters and were unrelated to clinicians' degree of confidence in their ratings. These results indicate a need to standardize methods for clinical assessment of bruxism. Additionally, they have implications for studies using clinical assessments of bruxism to test the association between bruxism and other conditions such as TMDs. PMID- 12535136 TI - Bonding abutments to cast metal post/cores: comparison of pre-treatment effects. AB - Bond strengths were evaluated for (1) metal primer systems when the metal was contaminated by a dentin conditioner and (2) a dentin adhesive system when dentin was contaminated by metal primers. Disc specimens were cast in a silver-palladium copper-gold (Ag-Pd-Cu-Au) alloy and dentin specimens were prepared by grinding the labial surface of bovine teeth. Specimens were treated with (1) metal primer alone, (2) dentin conditioner alone, (3) metal primer followed by dentin conditioner and (4) dentin conditioner followed by metal primer. A resin cement was poured into a mould over a restricted bonding area and allowed to set. Metal specimens were shear stressed to failure after thermocycling (4-60 degrees C; 20,000 cycles). Dentin specimens were stressed in the same manner after 24 h of immersion in 37 degrees C water. The results were compiled and analysed by anova. Data for dentin specimens treated with dentin conditioner only or with the combination of dentin conditioner and metal primer were not significantly different, statistically. Post-thermocycled groups indicated that bond strengths to the alloy significantly decreased (P < 0.05) when the primed metal surface was contaminated with dentin conditioner regardless of the timing of its application. PMID- 12535137 TI - Changes in surface roughness and colour stability of soft denture lining materials caused by denture cleansers. AB - Soft denture lining materials were immersed into solutions of denture cleansers for 8 h at room temperature, and immersed into distilled water for the remainder of the 24-h period at 37 degrees C. Surface roughness of the soft denture lining materials was measured by contact type surface roughness instrument. For the colour stability test, soft denture lining materials were immersed in the denture cleansers as described above for 180 days. Finally, the colour changes of each material were quantitatively measured by a photometrical instrument to obtain the colour differences between newly processed specimen and immersed specimens (P < 0.01). An autopolymerizing silicone material, Evatouch, exhibited severe changes in surface roughness by all denture cleanser, and the generic material GC Denture Relining showed the minimal changes. Severe colour changes were also observed with some liner and cleanser combinations (P < 0.01). Except for Evatouth, the four silicone soft liners were more stable in surface roughness and in colour change than the two acrylic soft liners. One autopolymerizing silicone (GC denture relining) and one heat curing silicone (Molloplast B) demonstrated the best stability. PMID- 12535138 TI - Longitudinal study of the influence of removable partial denture and chemical control on the levels of Streptococcus mutans in saliva. AB - Some studies have evaluated the salivary levels of mutans streptococci (MS) in removable partial denture (RPD) users. Saliva samples (2.0 mL) were obtained from 31 patients in six periods: (T0): immediately before installation of RPD; (T8): 8 days after T0; (T48): 48 days after T0; (T92): 92 days after T0; (T140): 140 days after T0 and (T189): 189 days after T0. The samples were vortexed and serially diluted from 10(-1) to 10(-6) in 0.05 m phosphate buffer (pH 7.4). From each dilution, 0.025 mL was plated on Mitis Salivarius Bacitracin (MSB). The plates were incubated in 5% CO2 at 37 degrees C for 72 h. There was an increase (t-test, P < 0.05) in the number of MS between periods T0 and T48 (mean/s.d., CFU mL(-1) of saliva): T0: 2.26/4.43 x 10(6) and T48: 0.47/1.48 x 10(8). After this, intensive treatment with CHX was accomplished in 29 patients. Saliva samples were obtained after treatment in four periods: (T24 h): 24 h after T0; (T14): 14 days after T24 h; (T28): 28 days after T24 h, and (T63): 63 days after T24 h. The number of MS in saliva did not decrease (t-test, P > 0.05). A new CHX formulation was applied in 15 patients. Saliva samples were obtained in periods: (T0): before new CHX application; (T24 h): 24 h after T0 and (T82): 82 days after T0. The new CHX reduced MS levels in saliva: (mean/s.d., CFU mL(-1) of saliva): T0: 6.64/8.47 x 10(6) and T24 h: 3.2/4.27 x 10(5) (sign rank, P < 0.05). In conclusion, there was a significant increase in the number of MS in saliva after the installation of RPD. The intensive treatment with a properly formulated CHX was effective in the reduction of MS, between 24 h and 82 days after its application. PMID- 12535139 TI - First night effect of an interocclusal appliance on nocturnal masticatory muscle activity. AB - The purpose of this study was to examine the effect of an interocclusal appliance on nocturnal masticatory muscle activities. Six healthy Japanese males (mean age: 26.8 years) participated in this study. Electromyographic (EMG) activities of the right anterior temporalis and masseter muscles were recorded using a portable EMG recording unit at night both with and without an interocclusal appliance. In both muscles, the maximal EMG activity and the number of bruxing events decreased significantly by wearing the appliance. Moreover, the duration of a higher level of muscle activity was decreased while that of a lower level of muscle activity increased by wearing the appliance in both muscles. These findings suggest that nocturnal masticatory muscle activity is significantly reduced by wearing an interocclusal appliance, and that the use of such an appliance at night could help to relax masticatory muscles. PMID- 12535140 TI - Validity and reliability of a newly developed method for evaluating masticatory function using discriminant analysis. AB - The objective of the study was to assess validity and reliability of a newly developed method for evaluating mixing ability of masticatory function. A two coloured paraffin wax cube was used as a test food in the mixing ability test. Eleven full dentate and 33 denture wearers participated in the validity assessment. They were divided into three groups according to their dental state. Validity was assessed by the Spearman's coefficient of correlations between the mixing ability test and the sieving method used as a 'gold standard' test of the masticatory performance. The subjects were asked to masticate the test foods for 5, 7, 10, 15, 20 and 30 strokes in each test. Significant correlations (P < 0.001) were obtained in natural dentition group (r(s) = 0.66), in denture-natural dentition group (r(s) = 0.72) and in denture-denture group (r(s) = 0.56). In the reliability assessment, three full dentate subjects were asked to repeat the mixing ability test a number of times. The intraclass correlation coefficients were 0.94 in interexaminer consistency, 0.98 in intra-examiner consistency, 0.89 in test-retest consistency and 0.99 in measuring consistency. These results indicated that the mixing ability test had the validity and the reliability for evaluation of the masticatory performance. PMID- 12535141 TI - Postnatal development of protein gene product 9.5 and calcitonin gene-related peptide immunoreactive nerve fibres in rat temporomandibular joint disc. AB - Protein gene product 9.5 (PGP 9.5), an immunohistochemical marker of whole nerve fibres, and calcitonin gene-related peptide (CGRP), a marker of thin nerve fibres, were used to elucidate the postnatal development of nerve fibres in rat temporomandibular joint (TMJ) disc. At birth, PGP 9.5-immunoreactive nerve fibres exhibited running towards the central area of the disc, invading by approximately 95 m from the disc attachment. The nerve fibres existing inside the disc became longer during postnatal development. The number of nerve fibres in the disc increased in a progressive manner up to 40 days after birth. CGRP-immunoreactive nerve fibres also presented changes essentially similar to those of PGP 9.5 immunoreactive nerve fibres. However, the proportion of CGRP-immunoreactive nerve fibres to PGP 9.5-immunoreactive ones was approximately 80%, and remained constant up to 40 days after birth. In conclusion, the distribution and the number of nerve fibres are variable during postnatal development, although the ratio of thin nerve fibres remains invariable. It is emphasized that these changes of innervation in the TMJ are associated with the development of masticatory function. PMID- 12535142 TI - The interaction of glass-ionomer cements containing vinylphosphonic acid with water and aqueous lactic acid. AB - A glass-ionomer cement containing an acrylic acid/vinyl phosphonic acid copolymer, has been investigated for its interaction with water and with aqueous lactic acid and the results compared with those from conventional glass-ionomers based on polyacrylic acid. Cylindrical specimens (12 mm high x 6 mm diameter) were placed in 8 cm3 of aqueous lactic acid (20 mmol dm(-3); pH 2.7) for 1 week, at the end of which the pH was determined. Each specimen was then placed in a fresh 8 cm3 volume of lactic acid and the pH determined after a further week. This procedure was continued for a total of 13 weeks for each specimen. Experiments were also carried out on similar specimens exposed to 8 cm3 of water for 1 week only. Further experiments were carried out in which discs of cement were exposed to thin films (0.15 mm) of lactic acid at pH 4.5, with pH values determined at 30 s, 1, 2, 5 and 10 min. After 1 week, cements had changed the pH of the lactic acid to a mean value of 3.63 (SD 0.08) while in weeks 2-13 they changed it to a mean value of 3.31 (SD 0.11). After 1 week in aqueous lactic acid the cements had gained mass by a mean of 1.26% (SD 0.59%) compared with 1.22% (SD 0.14%) in water. After 13 weeks in lactic acid, cements had lost a mean of 2.83% (SD 0.74%) in mass. Thin films of aqueous lactic acid changed from pH 4.5-5.1 at 30 s and to a steady value of 5.6 (SD 0.3) between 2 and 10 min. These results were similar to those for conventional glass-ionomer cements based on carboxylic acid polymers. Hence, it was concluded that the presence of the vinylphosphonic acid units made no significant difference to the interaction of cements with aqueous solutions. PMID- 12535143 TI - A clinical and radiographic evaluation of fixed partial dentures (FPDs) prepared by dental school students: a retrospective study. AB - In recent years, there has been an increasing interest in fixed partial dentures (FPD) quality, expected lifetime and possible reasons for failure. This study probed the FPD success rate and the relationship among various factors and their effect on FPD failure. Patients treated by dental students under supervision and installed FPDs were examined clinically and radiographically. A total of 247 FPDs were evaluated after a mean service period of 6.3 years (ranging from 1 to 25 years) after cementation. Caries was found to be the most frequent cause of failure and non-vital abutments decreased survival rates. PMID- 12535144 TI - Comparison of flexural properties of composite restoratives using the ISO and mini-flexural tests. AB - The purpose of this study was to investigate the flexural properties (flexural strength and flexural modulus) of four commercial composite restoratives (Silux Plus, Z100, Ariston and Surefil) using the ISO 4049 flexural test (IFT) and a mini-flexural test (MFT). Both tests involved the use of three-point loading and the same fixture. The difference between the tests was in the length of the composites specimens and the distance between the supports [20 mm (IFT) and 10 mm (MFT)]. Six specimens were made for each material and flexural test. Test specimens [25 x 2 x 2 mm (IFT) and 12 x 2 x 2 mm (MFT)] were fabricated according to manufacturers' recommendations. After light-polymerization, the specimens were stored in distilled water at 37 degrees C for 24 h. The specimens were subsequently blotted dry, measured and subjected to flexural testing using an Instron Universal Testing Machine with a crosshead speed of 0.75 mm min(-1). Data was analysed using anova/Scheffe's, paired samples test (P < 0.05) and Pearson's correlation (P < 0.01). For both IFT and MFT, results of statistical analysis of flexural strength were identical. Silux had significantly lower flexural strength compared with the other composites and the flexural strength of Ariston was significantly lower than Z100 and Surefil. For IFT, the flexural modulus of Z100 was significantly higher than Silux, Ariston and Surefil while for MFT, Silux had significantly lower modulus compared with Z100, Ariston and Surefil. A significant, strong and positive correlation (r = 0.95) was observed for flexural strength between IFT and MFT. Correlation for flexural modulus was also significant and positive but was weaker (r = 0.53). As MFT has the advantage of ease of specimen fabrication and is more clinically realistic, it is suggested for the testing of composite restoratives. CLINICAL RELEVANCE: The mini-flexural test may be better than the ISO flexural test for screening of composite restoratives for clinical applications. PMID- 12535145 TI - Effect of fluoride-releasing adhesive system on decalcified dentin. AB - The aim of this study was to evaluate the effect of fluoride-releasing adhesive systems on human decalcified dentin in vitro. Two fluoride-releasing adhesive systems, Reactmer bond (RB, Shofu) and ABF (AF, Kuraray), an experimental system, and a commercial adhesive system without fluoride release, SE bond (SE, Kuraray), were used in this study. The amount of fluoride release from adhesive in deionized water was measured every week for 10 weeks. Class V cavities were prepared on extracted human pre-molars and decalcified dentin was promoted by using a bacterial caries induction system at the cavity floor. The cavities preserving decalcified dentin were restored with resin composite (AP-X, Kuraray) after treatment by each adhesive system. The specimens without treatment by adhesive system and restoration were used for control. The specimens with restoration were then incubated for 4 weeks at 37 degrees C, 100% humidity. Microradiographs of the specimens showed that the radiopacities of the decalcified dentin layers in RB and AF groups with fluoride release were significantly higher than those in SE or control groups without fluoride release. This result suggested that the fluoride-releasing adhesive systems enhanced mineralization of decalcified dentin. PMID- 12535146 TI - Influence of abutment substrates on the colour of metal-free polymer crowns. AB - New-generation composite materials have excellent strength and wear resistance, and thus can be used to make metal-free crowns. However, composite materials are translucent, and so when using them to make metal-free crowns, it is necessary to take the effect of the abutments into consideration. In this study, five types of materials including three types of new-generation composite materials, a conventional composite material, and a ceramic material were used to examine the translucency (contrast ratio) of the materials and the effects of the colour of the abutments on the final appearance of metal-free crowns. It was found that the materials varied slightly from one-another in translucency and that it was possible to reproduce the desired colour when gold alloy was used for the background. However, it was difficult to produce an adequate colour match when silver-palladium alloy was used for the background. When a tooth colour material that was darker than the target colour was used for the abutment teeth, some materials could not reproduce the target colour satisfactorily. PMID- 12535147 TI - Facial and glossal distribution of anaesthesia after inferior alveolar nerve block. AB - The aim of this study was to subjectively determine the distribution of anaesthesia by mapping areas of sensory loss following inferior alveolar nerve block. Fifty healthy dental students were the subjects of this study (men 32, women 18). They were asked to draw the anaesthetized area on a diagram of the face and tongue 20 min after inferior alveolar nerve block. They evaluated the degree of anaesthesia by touching their faces and moving their tongues. All of the 50 subjects reported anaesthesia in the facial area. Of these, 21 (42%) reported the cutaneous distribution of anaesthesia on mental nerve territory only. Seventeen subjects (34%) reported anaesthesia on mental and buccal nerve territory. Nine subjects (18%) reported anaesthesia on mental, buccal, and auriculotemporal nerve territory. Two subjects (4%) reported anaesthesia on mental and auriculotemporal nerve territory and one subject (2%) on mental, buccal and infra-orbital nerve territory. Forty-seven of the 50 subjects (94%) reported anaesthesia of the tongue with the various degree of anaesthesia according to the area. Of these, 17 subjects (34%) reported strong anaesthesia on the anterior area and weak anaesthesia on the middle part of the tongue. Nineteen subjects (38%) reported strong anaesthesia of the lateral area and weak anaesthesia on the medial area, and 11 subjects (22%) reported anaesthesia on only the lateral side of the tongue. Three subjects (6%) reported no anaesthesia of the tongue. The distribution of anaesthesia of the facial and glossal regions determined subjectively after inferior alveolar nerve block, varies significantly between individuals. PMID- 12535148 TI - Evaluation of alternative intra-oral repair techniques for fractured ceramic fused-to-metal restorations. AB - Ceramic fractures are serious and costly problems in dentistry. Moreover, they pose an aesthetic and functional dilemma both for the patient and the dentist. This problem has created demand for the development of practical repair options which do not necessitate the removal and remake of the entire restoration. Published literature on repair techniques for fractured fixed partial dentures, concentrating on the data obtained both from in vitro and in vivo studies, reveals that the repair techniques based on sandblasting and silanization are the most durable in terms of adhesive and cohesive failures compared with those using different etching agents. PMID- 12535149 TI - A comparison of the erosive susceptibility of cuspal and cervical enamel using an ultrasonic technique. AB - The aim of this study was to compare the erosive susceptibility of cuspal and cervical enamel from human premolar and molar teeth. Small blocks of cervical and cuspal enamel were immersed in either orange juice or Coca-Cola at 37 degrees C and the surface enamel loss was measured by surfometry at 1, 2, 3 and 4 h. Additionally, once-hourly enamel loss was measured, specimens were placed in an ultrasonic bath containing water and ultrasonicated for 5 s to determine the degree of surface demineralization. A further set of enamel specimens were prepared that had 100 microm of the enamel surface removed. This was done to remove the hypermineralized surface enamel layer. Surface enamel loss in orange juice at 4 h, following ultrasonication, ranged from 13.2 to 16.9 microm. The surface enamel loss in Coca-Cola at 4 h, following ultrasonication, ranged from 21.7 to 27.5 microm. Subsurface enamel loss in orange juice at 4 h, following ultrasonication, ranged from 10.7 to 16.1 microm. The subsurface enamel loss in Coca-Cola for 4 h, following ultrasonication, ranged from 36.8 to 37.2 microm. Overall, little difference was found in the erosive susceptibility of cervical and cuspal enamel to the effects of orange juice or Coca-Cola. PMID- 12535150 TI - The influence of war stress on the prevalence of signs and symptoms of temporomandibular disorders. AB - The aim of this study was to determine the prevalence of signs and symptoms of temporomandibular disorders in patients with post-traumatic stress disorder (PTSD) occurring as a consequence of exposure to war stress. The examined group consisted of 100 male subjects, aged from 25 to 50 years, who had taken part in the War in Croatia, and in whom PTSD had previously been diagnosed. The control group comprised 100 subjects, without PTSD, of the same sex and age group, who had not taken part in the War. A functional evaluation of the stomatognathic system was performed using the Helkimo Anamnestic and Clinical Dysfunction Index. Eighty-two per cent of the group with PTSD had at least one symptom, and 98% at least one sign of dysfunction. Eight per cent reported mild symptoms and 74% severe symptoms. Twenty-eight per cent showed mild clinical signs, 22% had signs of moderate, and 48% of severe dysfunction. Twenty-four per cent of the control group of subjects had at least one symptom, and 52% at least one clinical sign of dysfunction. Twenty-two per cent reported mild and 2% severe symptoms. Mild clinical signs of dysfunction were found in 50% of subjects in the control group, moderate in 2% and none exhibited severe clinical signs. It is concluded that correlation exists between war stress and temporomandibular disorders. PMID- 12535151 TI - Effect of finishing and polishing procedures on the surface roughness of new tooth-coloured restoratives. AB - The purpose of this study was to evaluate the effect of various finishing and polishing procedures on the surface roughness of three new tooth-coloured restorative materials. The materials included a hybrid composite resin (Filtek Z250), a packable composite resin (Filtek P60), and an ormocer (organically modified ceramic) (Definite). Thirty-two specimens of each material were prepared and randomly divided into four finishing and polishing groups (n = 8): (i) diamond bur/silicone polishers, (ii) diamond bur/Sof-Lex discs, (iii) carbide bur/silicone polishers, (iv) carbide bur/Sof-Lex discs. Average surface roughness (Ra) in micrometers was measured with a Mitutoyo Surftest-402 Surface Roughness Tester and the data compared using anova, at P < or = 0.05. Surface topography was also assessed using the scanning electron microscope (s.e.m) from samples of each group. The Mylar strip produced the smoothest surface and finishing/polishing procedures; (ii) and (iv) were significantly smoother than (i) and (iii). The lowest variability in initial surface roughness between materials was also observed with ormocer group. PMID- 12535153 TI - In vivo gene transfer studies on the regulation and function of the vasopressin and oxytocin genes. AB - Novel genes can be introduced into the germline of rats and mice by microinjecting fertilized one-cell eggs with fragments of cloned DNA. A gene sequence can thus be studied within the physiological integrity of the resulting transgenic animals, without any prior knowledge of its regulation and function. These technologies have been used to elucidate the mechanisms by which the expression of the two genes in the locus that codes for the neuropeptides vasopressin and oxytocin is confined to, and regulated physiologically within, specific groups of neurones in the hypothalamus. A number of groups have described transgenes, derived from racine, murine and bovine sources, in both rat and mouse hosts, that mimic the appropriate expression of the endogenous vasopressin and genes in magnocellular neurones (MCNs) of the supraoptic and paraventricular nuclei. However, despite considerable effort, a full description of the cis-acting sequences mediating the regulation of the vasopressin-oxytocin locus remains elusive. Two general conclusions have nonetheless been reached. First, that the proximal promoters of both genes are unable to confer any cell specific regulatory controls. Second, that sequences downstream of the promoter, within the structural gene and/or the intergenic region that separates the two genes, are crucial for appropriate expression. Despite these limitations, sufficient knowledge has been garnered to specifically direct the expression of reporter genes to vasopressin and oxytocin MCNs. Further, it has been shown that reporter proteins can be directed to the regulated secretory pathway, from where they are subject to appropriate physiological release. The use of MCN expression vectors will thus enable the study of the physiology of these neurones through the targeted expression of biologically active molecules. However, the germline transgenic approach has a number of limitations involving the interpretation of phenotypes, as well as the large cost, labour and time demands. High-throughput somatic gene transfer techniques, principally involving the stereotaxic injection of hypothalamic neuronal groups with replication-deficient adenoviral vectors, are now being developed that obviate these difficulties, and which enable the robust, long-lasting expression of biologically active proteins in vasopressin and oxytocin MCNs. PMID- 12535152 TI - The effects of the phytoestrogen, coumestrol, on gonadotropin-releasing hormone (GnRH) mRNA expression in GT1-7 GnRH neurones. AB - Phytoestrogens can produce inhibitory effects on gonadotropin secretion in both animals and humans, although little is known about the mechanisms and the role of direct action on oestrogen receptors (ER) in this process. We examined the effect of coumestrol, alone and combined with ER antagonists, on gonadotropin-releasing hormone (GnRH) mRNA expression in GT1-7 cells. Coumestrol was found to have an inhibitory effect compared to controls, which was blocked by R,R-THC, a selective ER beta antagonist. These results suggest that ER beta is involved in the suppression of GnRH mRNA expression by coumestrol. PMID- 12535154 TI - Interleukin-1 beta depolarizes paraventricular nucleus parvocellular neurones. AB - Interleukin-1 beta (IL-1 beta) is involved in hypothalamic regulation of corticotropin releasing hormone (CRH) secretion and consequent downstream modulation of the neuroimmune response. In this study, whole-cell patch clamp recordings of rat parvocellular neurones in a slice preparation of the paraventricular nucleus (PVN) of the hypothalamus were performed to examine the cellular effects of IL-1 beta. In response to 1 nm IL-1 beta, 65% of parvocellular neurones tested exhibited a clear depolarization, which was abolished in the presence of tetrodotoxin (TTX). This depolarization was partially dependent on nitric oxide formation, as demonstrated by attenuation of the response in the presence of N-omega-nitro-L-arginine methylester, a nitric oxide synthase inhibitor. The effects of IL-1 beta on responsive parvocellular neurones were associated with a decrease in the frequency of inhibitory post synaptic potentials (IPSPs). Bicuculline administration blocked the effects of IL 1 beta, suggesting that this cytokine modulates GABA-ergic output, resulting in a decrease in inhibitory input (IPSPs) and consequent depolarization. These data support the conclusion that IL-1 beta influences the excitability of parvocellular neurones in the PVN, as a secondary consequence of nitric oxide generation and modulation of GABAergic inhibitory input to these cells. They elucidate cellular correlates underlying the well-established neuroimmune roles of IL-1 beta in the paraventricular nucleus of the hypothalamus. PMID- 12535155 TI - Bisphenol A increases progesterone receptor immunoreactivity in the hypothalamus in a dose-dependent manner and affects sexual behaviour in adult ovariectomized rats. AB - Recently, we reported that bisphenol A (BPA), an endocrine disrupter, increased progesterone receptor (PR) mRNA in the preoptic area (POA) in adult ovariectomized rats. In the present study, we examined whether BPA also induced expression of PR proteins in both the POA and the ventromedial hypothalamic nucleus (VMH), and whether those proteins were involved in the induction of sexual behaviour. Two weeks after ovariectomy, rats received a subcutaneous (s.c.) injection of BPA, 17 beta-oestradiol or vehicle. Twenty-four hours after the injection, the rats were killed and their tissues were examined by immunocytochemistry. Some rats that received a s.c. injection of BPA, E2 or vehicle alone on the day before were injected with progesterone at 15.00 h and examined for sexual behaviour 5-7 h later. As expected, injection of 10 microg E2 significantly increased the number of PR immunoreactive cells in both the POA and the VMH compared to the number after injection of vehicle alone. In both the POA and the VMH, injection of BPA at a dose of 10 mg also significantly increased the number of PR immunoreactive cells compared to the number after injection of sesame oil alone. Furthermore, BPA induced a dose-dependent increase in the number of PR immunoreactive cells in both the POA and the VMH, demonstrating that the number of PR cells was significantly increased by as little as 100 microg of BPA. Ovariectomized (OVX) rats that were primed with 10 mg of BPA, followed by 1 mg of progesterone, displayed mainly rejection behaviour, but not lordosis as typically observed in OVX rats primed with E2 followed by progesterone. The present study suggests that BPA influences reproductive functions, including sexual behaviour even in adulthood, by altering the PR system in the hypothalamus. PMID- 12535156 TI - Lipopolysaccharide endotoxin potentiates the effect of osmotic stimulation on vasopressin synthesis and secretion in the rat hypothalamus. AB - Vasopressin secreted by magnocellular neurones of the hypothalamic supraoptic and paraventricular nuclei is essential for water balance. In this study, we examined magnocellular neurone responses to osmotic stimulation in vehicle-injected controls or rats receiving an intraperitoneal (i.p.) injection of 250 microg/100 g of lipopolysaccharide (LPS), 3 h or 6 h earlier. LPS injection had no effect on plasma vasopressin concentrations in control rats but it caused marked and transient potentiation of the responses to a single i.p. injection of hypertonic saline (five- and two-fold, 3 and 6 h after LPS, respectively). The enhancement of plasma vasopressin responses was independent of plasma sodium concentrations or changes in blood pressure. Basal vasopressin mRNA expression in the paraventricular and supraoptic nuclei decreased slightly 6 h after LPS injection, without changes in vasopressin transcription as indicated by vasopressin heteronuclear (hn) RNA levels. Parvocellular neurones showed expected increases in vasopressin hnRNA expression following LPS injection and a further increase after i.p. hypertonic saline injection (due to the painful component). In contrast to magnocellular vasopressin mRNA expression, the effects of LPS and hypertonic saline injections in parvocellular neurones were additive and not synergistic. Light microscopic immunohistochemical examination revealed an increase in size of vasopressin but not oxytocin axonal terminals in the neural lobe 3 h after LPS injection. Osmotic stimulation caused marked depletion of vasopressin immunoreactivity in axonal terminals of the neural lobe in both control and LPS-pretreated rats. The changes in vasopressin axon terminals were accompanied by induction of interleukin (IL)-1 beta and IL-6 in the posterior pituitary. The data show that endotoxemia causes morphological and functional alterations of the hypothalamic neurohypophyseal system, resulting in facilitation rather than inhibition of vasopressin synthesis, and secretion in response to osmotic stimulation. PMID- 12535157 TI - Effects of acute treatment with 8-OH-DPAT and fluoxetine on aggressive behaviour in male song sparrows (Melospiza melodia morphna). AB - The role of serotonin in modulating male aggressive behaviour was investigated in male song sparrows, Melospiza melodia morphna, using two different serotonergic drugs, fluoxetine and 8-OH-DPAT. Fluoxetine is a selective serotonin reuptake inhibitor of the neuronal reuptake pump increasing synaptic concentrations of serotonin, and 8-OH-DPAT is a specific serotonin (5-HT1A) receptor agonist. The serotonergic control of aggression in passerines has not been previously investigated. We examined these behaviours within a controlled setting using a laboratory simulated territorial intrusion, with a hierarchical scale to quantify male-male aggressive behaviour. Utilizing this scale, we quantified the extent of male aggressive behaviour in two experiments. In experiment 1, song sparrows were given 100 micro l, s.c. injections of either fluoxetine (10 mg/kg) or 8-OH-DPAT (1 mg/kg). Experiment 2 was a dose-response study using three doses of 8-OH-DPAT (0.1, 1 and 10 mg/kg). In both studies, aggressive behaviour was measured 1 h after injection for 10 min in response to the presence of a novel male decoy combined with playback of conspecific song. Both drugs significantly reduced male aggressive behaviour, and 8-OH-DPAT did so in a dose-dependent manner. The effect of the two drugs upon general activity was also measured using infra-red perch hop detectors. Activity levels were not effected by either fluoxetine or 8-OH DPAT at all of the respective doses, indicating that the reduction in aggressive behaviour was specific. These results demonstrate that, in a passerine species, the serotonergic system negatively regulates male-male aggressive behaviour. These results further demonstrate that aggression can be effectively studied in a laboratory setting and natural aggressive responses can be elicited using this method. PMID- 12535158 TI - Photoperiodic and temporal influences on chemosensory induction of brain fos expression in female prairie voles. AB - Prairie voles (Microtus ochrogaster) typically stop breeding during winter. Male prairie voles respond to winter day lengths with gonadal regression, whereas female voles are relatively unresponsive to photoperiod. Unlike commonly studied laboratory rodents, female prairie voles do not exhibit spontaneous oestrous cycles. Instead, females are induced into oestrus by chemosensory cues from conspecific male urine. The present study investigated the interaction among day length, chemosensory cues and the initial brain responses during oestrus induction in female voles. A single drop of male conspecific urine, saline or skimmed milk was applied to the nares of female prairie voles housed for 9 weeks in either long (LD 16 : 8 h) or short (LD 8 : 16 h) days. Animals were killed 0.5, 1, 2 or 24 h after chemosensory treatment and their brains were processed for Fos immunocytochemistry. Body mass and ovarian fat pad mass were higher, but uterine and ovarian mass were lower, in short-day compared to long-day females. Regardless of photoperiod, Fos- immunoreactivity increased in the granule layer of the accessory olfactory bulb (AOB), the supraoptic nucleus and bed nucleus of the stria terminalis (BNST) (anterior medial) in females treated with male urine compared to the two control groups. Fos staining intensified in the AOB, medial and posterocortical medial amygdala and BNST (posterior ventral), 1 h and 2 h after urine treatment. In the medial preoptic area, anterior and lateral hypothalamus, and ventromedial nucleus of the hypothalamus, Fos-immunoreactivity was elevated in females 2 h after receiving urine. Overall, long-day females displayed higher Fos expression in response to urine than females maintained in short days. These results identify a putative neural circuitry of oestrus induction in this species, and provide an approximate time line of activation in the brain circuit responsible for oestrus induction in prairie voles. PMID- 12535159 TI - The melanocortin agonist Melanotan-II reduces the orexigenic and adipogenic effects of neuropeptide Y (NPY) but does not affect the NPY-driven suppressive effects on the gonadotropic and somatotropic axes in the male rat. AB - Neuropeptide Y (NPY) is a strong orexigenic neurotransmitter also known to modulate several neuroendocrine axes. alpha-Melanocyte-stimulating hormone (MSH) is an essential anorectic neuropeptide, acting on hypothalamic MC3/4 receptor subtypes. When given as an intracerebroventricular bolus injection, Melanotan-II (MT-II), a non selective MC receptor agonist, inhibits feeding, suppresses the NPY orexigenic action, and reduces basal insulinaemia. We evaluated the effects of a 7-day central infusion of MT-II (15 nmol/day) given either alone or in association with NPY (5 nmol/day) in male Sprague-Dawley rats. MT-II produced almost full anorexia for 1-2 days but then feeding gradually returned to normal despite continued MT-II infusion. When coinfused with NPY, MT-II also produced the same initial anorectic episode but then maintained feeding to upper normal levels, thus cancelling the hyperphagia driven by NPY. Whereas NPY infusion produced a doubling of fat pad weight, MT-II reduced adiposity by a factor of two compared to pair-fed rats, and vastly curtailed the NPY-driven increase in fat pad weight. MT-II infusion also significantly curtailed the NPY-induced rise in insulin and leptin secretions. NPY infusion significantly inhibited hypothalamic pro-opiomelanocortin mRNA expression, most likely cancelling the alpha-MSH anorectic activity. As expected from previous studies, chronic NPY infusion strongly inhibited both the gonadotropic and somatotropic axes, and coinfusion of MT-II did not reverse these NPY-driven effects, in sharp contrast with that seen for the metabolic data. MT-II infusion alone had little effect on these axes. In conclusion, chronic MT-II infusion generated a severe but transient reduction in feeding, suggesting an escape phenomenon, and clearly reduced fat pad size. When coinfused with NPY, MT-II was able to cancel most of the NPY effects on feeding, but not those on the neuroendocrine axes. It appears therefore that, as expected, NPY and alpha-MSH closely interact in the control of feeding, whereas the neural pathways by which NPY affects growth and reproduction are distinct and not sensitive to MC peptide modulation. PMID- 12535160 TI - Vasopressin and oxytocin decrease excitatory amino acid release in adult rat supraoptic nucleus. AB - Oxytocin and vasopressin reduce the amplitude of excitatory postsynaptic responses in magnocellular neuroendocrine cells of the supraoptic nucleus (SON). To test whether synaptic glutamate release is modulated by these neuropeptides, we examined the combined effect of vasopressin and oxytocin on depolarization induced glutamate and aspartate release from acutely dissected rat SON or fronto parietal cortex punches. Glutamate release was stimulated with 60 mm K+ for 5-10 min and measured using ion exchange chromatography or high-performance liquid chromatography. During depolarization with high K+, extracellular glutamate levels increased, on average, to 204% of control values. In the presence of vasopressin/oxytocin, K+-stimulated glutamate and aspartate release were significantly reduced by 34% and 62%, respectively, in the SON. Treatment with the aminopeptidase inhibitor amastatin did not mimic the effects of exogenous vasopressin/oxytocin on glutamate or aspartate release, suggesting that, under the conditions tested here, amastatin treatment may produce more complex effects. The effects of exogenous neuropeptides are likely mediated by oxytocin and/or vasopressin receptors, as the oxytocin- and V1a-receptor antagonist, Manning Compound (10-100 micro m), partially reversed the effects of vasopressin/oxytocin on SON glutamate release. In contrast, in cortical punches, glutamate release was enhanced by high K+, but vasopressin/oxytocin did not significantly reduce glutamate/aspartate release, consistent with the relatively sparse distribution of vasopressin/oxytocin receptors in fronto-parietal cortex. These findings suggest that locally released oxytocin and vasopressin may autoregulate SON magnocellular neuroendocrine cell activity in part by modulating the release of excitatory amino acids from afferent terminals targeting these cells and/or from other cellular sources. PMID- 12535161 TI - Termination of neuroendocrine refractoriness to melatonin in Siberian hamsters (Phodopus sungorus). AB - Siberian hamsters maintained from birth in a short day length (DL), unlike their long-day counterparts, fail to undergo reproductive development by 5 weeks of age. Instead, reproductive maturation of short-day males is delayed for approximately 20 weeks, at which point neuroendocrine refractoriness to the inhibitory effects of short DLs develops, resulting in growth of the gonads. To terminate refractoriness and re-establish responsiveness to short photoperiods, 10-15 weeks of long-day exposure is required. We assessed whether continuous exposure to long days is necessary to terminate refractoriness or whether the first few weeks of long days initiate a process that culminates several months later in the breaking of refractoriness. Male hamsters refractory to short DLs were transferred to a long-day photoperiod, pinealectomized (PINx) after 0, 3, 6 or 15 weeks, and subsequently infused for 6 weeks with a short-day melatonin signal. This melatonin treatment induces gonadal regression in photosensitive but not in photorefractory hamsters. Six percent of males PINx at week 0 and 88% of those PINx at week 15 underwent gonadal atrophy by the end of the melatonin infusion treatment initiated on week 15. Among hamsters PINx on week 6, 17% versus 76% underwent testicular involution in response to melatonin infusions initiated on week 6 and week 15, respectively. This finding indicates that a fraction of the long days that hamsters experience during spring and summer are sufficient to trigger the processes that restore responsiveness to short DLs. Additional groups of pineal-intact photorefractory animals were given 3, 6 or 15 weeks of long-day exposure and then returned to a short DL for several months; only those treated for 15 weeks terminated refractoriness. The breaking of refractoriness, once triggered by long-day melatonin signals, proceeds to completion only in the absence of short-day melatonin signals. PMID- 12535162 TI - Effect of intracerebroventricular administration of the octadecaneuropeptide on the expression of pro-opiomelanocortin, neuropeptide Y and corticotropin releasing hormone mRNAs in rat hypothalamus. AB - Intracerebroventricular (i.c.v.) administration of the octadecaneuropeptide (diazepam-binding inhibitor [33-50]; ODN) exerts a potent anorexigenic effect in the rat. We studied the effect of ODN on three neuropeptides involved in feeding behaviour: the orexigenic peptide neuropeptide Y (NPY) and two anorexigenic peptides, corticotropin-releasing hormone (CRH) and the pro-opiomelanocortin (POMC)-derived peptide alpha-melanocyte-stimulating hormone. The effect of i.c.v. administration of ODN (0.1 microg/kg and 1 microg/kg) on mRNA expression of the peptides in male rat hypothalamus was evaluated by semiquantitative in situ hybridization. In the arcuate nucleus, NPY-expressing neurones were mostly found in the inner zone in close proximity of the third ventricle. ODN at the dose of 0.1 microg/kg induced a significant decrease of 17.4% in NPY mRNA expression, while the depressing effect was more marked (31.4%) with the highest dose of ODN (1 microg/kg). POMC-expressing neurones were more laterally located in the arcuate nucleus. Administration of ODN at 0.1 microg/kg and 1 microg/kg doses induced increases of 33.5% and 27.4% in POMC mRNA expression, respectively. Labelling obtained with the CRH cRNA probe was essentially distributed throughout the medial parvocellular area of the hypothalamic paraventricular nucleus. ODN, at doses of 0.1 and 1 microg/kg, resulted in 17.8% and 32.8% decreases in CRH mRNA expression, respectively. The present data suggest that ODN might exert its anorexigenic effect by increasing mRNA expression of POMC and decreasing mRNA expression of NPY in the arcuate nucleus. PMID- 12535163 TI - A gonadotropin-releasing hormone insensitive, thapsigargin-sensitive Ca2+ store reduces basal gonadotropin exocytosis and gene expression: comparison with agonist-sensitive Ca2+ stores. AB - We examined whether distinct Ca2+ stores differentially control basal and gonadotropin (GTH-II)-releasing hormone (GnRH)-evoked GTH-II release, long-term GTH-II secretion and contents, and GTH-II-beta mRNA expression in goldfish. Thapsigargin (Tg)-sensitive Ca2+ stores mediated neither caffeine-evoked GTH-II release, nor salmon (s)GnRH- and chicken (c)GnRH-II-stimulated secretion; the latter responses were previously shown to involve ryanodine (Ry)-sensitive Ca2+ stores. Surprisingly, Tg decreased basal GTH-II release. This response was attenuated by prior exposure to sGnRH and caffeine, but was insensitive to the phosphatase inhibitor okadaic acid, the inhibitor of constitutive release brefeldin A and cGnRH-II. GTH-II-beta mRNA expression was decreased at 24 h by 2 microm Tg, and by inhibiting (10 microm Ry) and stimulating (1 nm Ry) Ry receptors. Transient increases in GTH-II-beta mRNA were observed at 2 h and 12 h following 10 microm and 1 nm Ry treatment, respectively. Effects of Tg, Ry and GnRH on long-term GTH-II secretion, contents and apparent production differed from one another, and these changes were not well correlated with changes in GTH II-beta mRNA expression. Our data show that GTH-II secretion, storage and transcription can be independently controlled by distinct Ca2+ stores. PMID- 12535164 TI - Chemical coding of GABA(B) receptor-immunoreactive neurones in hypothalamic regions regulating body weight. AB - Gamma-aminobutyric acid (GABA) interacts with hypothalamic neuronal pathways regulating feeding behaviour. GABA has been reported to stimulate feeding via both ionotropic GABA(A) and metabotropic GABA(B) receptors. The functional form of the GABA(B) receptor is a heterodimer consisting of GABA(B) receptor-1 (GABA(B)R1) and GABA(B) receptor-2 (GABA(B)R2) proteins. Within the heterodimer, the GABA-binding site is localized to GABA(B)R1. In the present study, we used an antiserum to the GABA(B)R1 protein in order to investigate the cellular localization of GABA(B)R1-immunoreactive neurones in discrete hypothalamic regions implicated in the control of body weight. The colocalization of GABA(B)R1 immunoreactivity with different chemical messengers that regulate food intake was analysed. GABA(B)R1-immunoreactive cell bodies were found in the periventricular, paraventricular (PVN), supraoptic, arcuate, ventromedial hypothalamic, dorsomedial hypothalamic, tuberomammillary nuclei and lateral hypothalamic area (LHA). Direct double-labelling showed that glutamic acid decarboxylase (GAD) positive terminals were in close contact with GABA(B)R1-containing cell bodies located in all these regions. In the ventromedial part of the arcuate nucleus, GABA(B)R1-immunoreactive cell bodies were found to contain neuropeptide Y, agouti related peptide (AGRP) and GAD. In the ventrolateral part of the arcuate nucleus, GABA(B)R1-immunoreactive cell bodies were shown to contain pro-opiomelanocortin and cocaine- and amphetamine-regulated transcript. In the LHA, GABA(B)R1 immunoreactivity was present in both melanin-concentrating hormone- and orexin containing cell populations. In the tuberomammillary nucleus, GABA(B)R1 immunoreactive cell bodies expressed histidine decarboxylase, a marker for histamine-containing neurones. In addition, GAD and AGRP were found to be colocalized in some nerve terminals surrounding GABA(B)R1-immunoreactive cell bodies in the parvocellular part of the PVN. The results may provide a morphological basis for the understanding of how GABA regulates the hypothalamic control of food intake and body weight via GABA(B) receptors. PMID- 12535165 TI - Metyrapone-induced glucocorticoid depletion modulates tyrosine hydroxylase and phenylethanolamine N-methyltransferase gene expression in the rat adrenal gland by a noncholinergic transsynaptic activation. AB - The hypothalamic corticotropin-releasing hormone system and the sympathetic nervous system are anatomically and functionally interconnected and hormones of the hypothalamic-pituitary-adrenocortical axis contribute to the regulation of catecholaminergic systems. To investigate the role of glucocorticoids on activity of the adrenal gland, we analysed plasma and adrenal catecholamines, tyrosine hydroxylase (TH) and phenylethanolamine N-methyltransferase (PNMT) mRNA expression in rats injected with metyrapone or dexamethasone. Metyrapone-treated rats had significantly lower epinephrine and higher norepinephrine production than control rats. Metyrapone increased TH protein synthesis and TH mRNA expression whereas its administration did not affect PNMT mRNA expression. Dexamethasone restored plasma and adrenal epinephrine concentrations and increased PNMT mRNA levels, which is consistent with an absolute requirement of glucocorticoids for PNMT expression. Adrenal denervation completely abolished the metyrapone-induced TH mRNA expression. Blockage of cholinergic neurotransmission by nicotinic or muscarinic receptor antagonists did not prevent the metyrapone induced rise in TH mRNA. Finally, pituitary adenylate cyclase activating polypeptide (PACAP) adrenal content was not affected by metyrapone. These results provide evidence that metyrapone-induced corticosterone depletion elicits transsynaptic TH activation, implying noncholinergic neurotransmission. This may involve neuropeptides other than PACAP. PMID- 12535166 TI - Central administration of neuropeptide FF causes activation of oxytocin paraventricular hypothalamic neurones that project to the brainstem. AB - Neuropeptide FF (NPFF), a morphine modulatory peptide, is emerging as an important neuromodulator in the context of central autonomic and neuroendocrine regulation. NPFF immunoreactivity and receptors have been identified in discrete autonomic regions within the brain and spinal cord, including the hypothalamic paraventricular nucleus (PVN). In this study, we examined the effects of intracerebroventricular (i.c.v.) administration of NPFF on activation of chemically identified PVN neurones that project to the brainstem nucleus of the solitary tract (NTS). In conscious rats, i.c.v. NPFF at a dose of 10 micro g, but not 8 micro g, caused an increase in arterial blood pressure. Immunohistochemical analysis revealed a dose-dependent increase in activated (Fos positive) PVN neurones following i.c.v. NPFF administration compared to controls receiving i.c.v. saline. Activated PVN neurones were located predominantly in the parvocellular compartment of the nucleus with relatively few Fos positive cells in the magnocellular subdivision. Chemical identification of activated neurones revealed significant number of activated cells to be oxytocin positive, whereas only few vasopressin, tyrosine hydroxylase (TH) or corticotrophin-releasing factor (CRF) neurones were double-labelled. Injection of the retrograde tracer fluorogold into the NTS resulted in labelling of significant numbers of parvocellular oxytocin, but not vasopressin, TH or CRF, PVN neurones. We conclude that centrally administered NPFF stimulates brainstem-projecting oxytocin PVN neurones. Oxytocin released from terminals within the NTS oxytocin thus modulate the activity of ascending visceral autonomic pathways that synapse initially within the NTS. PMID- 12535167 TI - Mechanisms of cytosolic Ca2+ suppression by prostaglandin E2 receptors in rat melanotrophs. AB - We have previously reported that voltage-dependent Ca2+ (VDC) channels of rat melanotrophs are inhibited by prostaglandin E2 (PGE2). In this study, mechanisms involved in the inhibitory actions of PGE2 receptors of rat melanotrophs were analysed using reverse transcriptase-polymerase chain reaction (RT-PCR), Ca2+ imaging and whole-cell, patch-clamp techniques with recently developed EP agonists, each of which is selective for the known four subclasses of EP receptors (EP1-4). PGE2 reversibly suppressed the cytosolic Ca2+ concentration ([Ca2+]i). The maximum reduction in [Ca2+]i by PGE2 was comparable to that by dopamine or to that by extracellular Ca2+ removal. RT-PCR analysis of all four EP receptors revealed that EP3 and EP4 receptor mRNAs were expressed in the intermediate lobe. The effects of PGE2 to suppress [Ca2+]i were mimicked by the selective EP3 agonist, ONO-AE-248, whereas three other EP agonists, ONO-DI-004 (EP1), ONO-AE1-259 (EP2) and ONO-AE1-329 (EP4), had little or no effect on [Ca2+]i. All four G-protein activated inward rectifying K+ (GIRK) channel mRNAs were identified in intermediate lobe tissues by RT-PCR. Dopamine concentration dependently activated GIRK currents, whereas PGE2 did not activate GIRK currents, even at the concentration causing maximal inhibition of VDC channels. These results suggest that PGE2 acts on EP3 receptors to suppress Ca2+ entry of rat melanotrophs by selectively inhibiting VDC channels of these cells. We have compared the possible cellular and molecular mechanisms of inhibition by dopamine and PGE2. PMID- 12535168 TI - Further delineation of the sequences required for the expression and physiological regulation of the vasopressin gene in transgenic rat hypothalamic magnocellular neurones. AB - We have introduced transgenes into rats with a view to defining genomic regions that mediate the cell-specific and physiological regulation of the vasopressin gene. These transgenes consist of the rat vasopressin structural gene with a reporter inserted into exon III, flanked by different lengths of upstream and downstream sequences. 11-VCAT-3 is flanked by 11 kbp of upstream sequences and 3 kbp of downstream sequences. The previously described 5-VCAT-3 is flanked by 5 kbp of upstream and 3 kbp of downstream sequences. 3-VCAT-3 has 3 kbp of upstream and 3 kbp of downstream sequences, and 3-VCAT-0.2 is flanked by 3 kbp of upstream and 0.2 kbp of downstream sequences. All four transgenes elicit the same expression patterns; low basal expression is seen in the magnocellular supraoptic and paraventricular nuclei, and is negligible in the suprachiasmatic nucleus. Expression increases markedly in vasopressin magnocellular cells following dehydration. The sequences responsible for the cell-specific expression and physiological regulation of our transgenes thus reside within the confines of the smallest construct studied, 3-VCAT-0.2. PMID- 12535169 TI - Orexin-A immunoreactivity and prepro-orexin mRNA expression in hyperphagic rats induced by hypothalamic lesions and lactation. AB - Orexins are endogenous neuropeptides that potently facilitate appetite and food consumption. In the present study, we examined orexin immunoreactivity and prepro orexin mRNA expression in the lateral hypothalamus by immunohistochemistry and competitive reverse transcription-polymerase chain reaction (RT-PCR) methods in different models of hyperphagia in rats. Hyperphagia was induced by lesions of either the ventromedial hypothalamus (VMHL) or the paraventricular nucleus (PVNL), and we also compared lactating rats to nonlactating controls. Both VMHL and PVNL increased food intake and body weight compared to shams. On day 7 post lesion, serum leptin and insulin concentrations exhibited 3.2- and 2.8-fold increases in VMHL rats, and nonsignificant 1.8- and 1.8-fold increases in PVNL rats; there were significant decreases (48% and 33%) in lactating rats on day 12 postpartum compared to controls, respectively. Serum glucose concentrations were not significantly changed compared to controls in these rats. Quantification by image analysis suggests that VMHL significantly decreased the number and mean staining intensity of orexin-A immunoreactive neurones compared to those in the sham-lesioned group; while PVNL did not change orexin-A immunoreactivity. Competitive RT-PCR analysis showed that VMHL significantly decreased the prepro orexin mRNA expression compared to those in the sham-lesioned group, and PVNL did not change it. Lactating rats on days 11-12 of lactation had significantly greater number and mean staining intensity of orexin-A immunoreactive neurones, prepro-orexin mRNA expression food intake and body weight than nonlactating postpartum rats. Thus, changes in orexin-A immunoreactivity and prepro-orexin mRNA expression were not consistent between the hyperphagia models. These results suggest that the hyperphagia from VMHL or PVNL and lactating rats differ in their involvement of orexin-A, and the change in circulating leptin and insulin concentrations may be involved in the change of orexin-A immunoreactivity in these rats. PMID- 12535170 TI - Suppression of pulsatile luteinizing hormone secretion but not luteinizing hormone surge in leptin resistant obese Zucker rats. AB - The adipose tissue-derived hormone leptin may be a primary mediator linking nutritional status and reproduction. The present study used the leptin-resistant obese female Zucker rat to investigate whether leptin signalling is required for normal pulsatile luteinizing hormone (LH) secretion and/or generation of the LH surge. For the pulsatile LH secretion study, an indwelling atrial catheter was implanted and a low dose of oestrogen given as a subcutaneous implant to lean and obese ovariectomized (OVX) Zucker rats. One week following OVX, blood samples were collected every 10 min for 3 h during the morning. Plasma LH concentrations were measured by radioimmunoassay. For the LH surge study, lean and obese OVX rats were given a high dose of oestrogen as a subcutaneous implant. Two days later, rats were given progesterone at 09.00 h to induce a proestrus-like LH surge. Blood samples were collected from an indwelling atrial catheter throughout that and the following day and plasma LH concentrations were measured by radioimmunoassay. LH pulse amplitude and mean LH secretion were profoundly attenuated in obese Zucker rats compared with lean littermates, whereas LH pulse frequency was not significantly different between phenotypes. The opioid receptor antagonist naloxone did not affect the pattern of pulsatile LH secretion in obese rats, suggesting that leptin does not exert its facilitatory effects on LH secretion through an opioidergic pathway. Both lean and obese rats showed characteristic steroid-induced LH surges. It therefore appears that a leptin signal is required for generation of a normal pattern of pulsatile LH secretion, but is not a necessary component of the steroid-induced LH surge. PMID- 12535171 TI - Nociceptin/orphanin FQ content is decreased in forebrain neurones during acute stress. AB - We examined the effects of acute and chronic stress on neurotransmission of nociceptin/orphanin FQ (N/OFQ) in a variety of brain regions. Four groups of rats were exposed to chronic variable stress, and/or a single acute stress before decapitation. Group 1 served as unstressed controls. The rats in group 2 (chronic stress/no acute stress) were exposed to a 10-day regimen of chronic stress (two unpredictable stressors per day). These rats were decapitated 20 h after the last stressor. The rats in group 3 (no chronic stress/acute stress) were not exposed to chronic stress, but they were restrained for 30 min prior to decapitation. The rats in group 4 (chronic stress/acute stress) were chronically stressed for 10 days, and were then restrained prior to decapitation. Trunk blood was collected, and plasma adrenocorticotrophic hormone (ACTH) and corticosterone (CORT) were assayed by radioimmunoassay (RIA). The rats' brains were dissected, and N/OFQ content was measured by RIA in a variety of brain regions, and in spinal cord. Chronic stress exposure altered the hormonal responses to the acute stress exposure. In the rats that were exposed to chronic stress without acute stress (group 2), N/OFQ content did not differ from the content of the unstressed controls in any of the dissected brain regions. In the two groups that were stressed acutely just before decapitation (groups 3 and 4), N/OFQ content was decreased by 25-30% in the basal forebrain. Accordingly, the neuronal content of N/OFQ is decreased in basal forebrain neurones during acute stress exposure. In light of our previous finding that N/OFQ administration increases circulating ACTH and CORT concentrations, and augments hormonal responses to an acute stressor, the current finding raises the possibility that endogenous N/OFQ participates in neuronal regulation of hormonal responses to acute stress exposure. PMID- 12535172 TI - Cerebral insulin increases brain response to glucose. AB - The hypothalamus participates in the regulation of carbohydrate metabolism involving a feedback loop between the brain and the periphery in which glucose sensitive hypothalamic areas appear to be involved. We have previously shown that a glucose injection (9 mg/kg) in the carotid artery toward the brain, in an amount that did not modify glycaemia, caused a rapid and transient increase in plasma insulin concentrations. To determine whether central insulin could influence this response, we investigated the change in central glucose-induced insulin secretion in intracerebroventricular (i.c.v) insulin-injected rats and in hyperinsulinaemic obese Zucker rats. Central glucose-induced insulin secretion was increased by 50% in i.c.v. insulin-injected rats compared to control rats. When a similar test was performed at a lower dose of glucose (3 mg/kg), a significant insulin secretion was observed only in rats submitted to a prior central insulin injection. These data indicate an increase in the brain response to glucose after insulin treatment. Using an identical lower glucose dose, we also demonstrated an enhanced brain glucose sensitivity in hyperinsulinaemic and insulin-resistant obese Zucker rats. Together, these results indicate that acute i.c.v. insulin or pathological hyperinsulinaemic state (i.e. obese Zucker rat) modulates the nervous control of insulin secretion by increasing the brain response to glucose. PMID- 12535173 TI - Central cholecystokinin-octapeptide accelerates the activity of the hypothalamic gonadotropin-releasing hormone pulse generator in goats. AB - To clarify central actions of cholecystokinin-octapeptide (CCK-8) on reproduction, effects of an intracerebroventricular (i.c.v.) administration of CCK-8 on the activity of the gonadotropin-releasing hormone (GnRH) pulse generator were examined in ovariectomized (OVX) goats in the absence or presence of oestradiol. Goats were chronically fitted with recording electrodes in the mediobasal hypothalamus, and electrophysiological manifestations of the GnRH pulse generator were monitored as characteristic increases in the multiple-unit activity (MUA volleys). In OVX goats, a bolus i.c.v. injection of as little as 0.01 nmol of CCK-8 induced a MUA volley with a short latency, which resulted in a significant decrease in the post-treatment volley interval compared to that in the saline injected control. Administration of higher doses of CCK-8 (0.1 and 2 nmol) did not further accelerate the occurrence of the MUA volley, but stimulatory effects were observed for a longer period than that after the 0.01 nmol injection. When goats were treated with oestradiol, while a bolus i.c.v. injection of 0.01 nmol CCK-8 had no effect, an injection of 0.1 nmol of the peptide significantly decreased the post-treatment volley interval. On continuous i.c.v. infusion of CCK-8 at 3 nmol per 200 micro l/h for 3 h, MUA volleys with shorter intervals than those in the control were successively induced without any apparent change in basal plasma luteinizing hormone levels in OVX goats. These results demonstrate that central CCK-8 strongly accelerates the activity of the GnRH pulse generator in goats. PMID- 12535174 TI - Thyroid hormone regulation of N-methyl-D-aspartic acid receptor subunit mRNA expression in adult brain. AB - Thyroid hormone is an essential modulator of brain development, but little is known about its actions in the adult brain. Hypothyroidism is associated with gene expression changes in both central and peripheral nervous tissue. Functional consequences of adult-onset hypothyroidism include an inability to produce long term potentiation in rat hippocampus and impaired learning and memory in both rats and man. Long-term potentiation is a form of learning that is dependent on functional N-methyl-d-aspartic acid (NMDA)-preferring ionotropic glutamate receptors. This work examines the expression of ionotropic glutamate receptor subunit mRNA following surgical thyroidectomy with or without thyroid hormone replacement. In situ hybridization histochemistry was used to determine the mRNA levels of the NMDA receptor subunits NR1, NR2A, NR2B, the AMPA receptor subunit GluR1, and the kainate receptor subunit KA2. Reducing circulating concentrations of thyroid hormone by surgical removal of the thyroid gland 2 weeks before sacrifice decreased the expression of NR1 mRNA exclusively in the hippocampus. Conversely, hyperthyroidism selectively reduced NR2B mRNA expression in the dorsal hippocampus. Altering thyroid hormone status had no effect on the expression of KA2 or GluR1 subunit mRNA. The regulation of expression of NR1 and NR2B mRNA by thyroid hormone is a novel mechanism for explaining the relationship between thyroid hormone and cognitive function. PMID- 12535175 TI - The anti-inflammatory effect of bee venom stimulation in a mouse air pouch model is mediated by adrenal medullary activity. AB - Cutaneous electrical or chemical stimulation can produce an anti-inflammatory effect, which is dependent on adrenal medullary-sympathetic activation. We have previously shown that peripheral injection of bee venom (BV) also produces a significant anti-inflammatory effect that is neurally mediated. In the present study, we examined whether this anti-inflammatory effect is also dependent on the adrenal gland using the mouse inflammatory air pouch model. Subcutaneous (s.c.) BV injection produced a marked suppression of leucocyte migration and tumour necrosis factor (TNF)-alpha concentration induced by zymosan injection into the air pouch. The role of the adrenal gland in this suppression was evaluated in adrenalectomized mice. Adrenalectomy significantly reversed the suppression of leucocyte migration and TNF-alpha elevation caused by BV. Serum concentrations of corticosteroid were increased in mice with zymosan-induced air-pouch inflammation and this increase was reduced by BV administration, suggesting that adrenal corticosteroid release is not involved in mediating the anti-inflammatory effects of BV. To test this hypothesis, the corticosteroid receptor antagonist (RU486) was administered and found not to affect the BV-induced inhibition of leucocyte migration. By contrast, pretreatment with the beta-adrenergic antagonist propranolol reversed the BV-induced inhibitory effect on leucocyte migration. These results suggest that the anti-inflammatory effect of s.c. BV administration is mediated in part by the release of catecholamines from the adrenal medulla. PMID- 12535176 TI - Temperature influences the ontogenetic expression of aromatase and oestrogen receptor mRNA in the developing tilapia (Oreochromis mossambicus) brain. AB - Water temperature has a differential influence on the development of central neurotransmitter systems according to the developmental period in tilapia (Oreochromis mossambicus). Aromatase and oestrogen receptors (ERs) represent important components of the mechanism of brain differentiation. Gene expression of aromatase and ERs is modulated by neurotransmitters in the developing brain. In the present study, the quantitative reverse transcription-polymerase chain reaction method was used to investigate the effects of temperature on the ontogenetic expression of aromatase and ERs in the developing tilapia brain. Before day 10 posthatching, exposure to a higher temperature (32 degrees C) resulted in a significant increase in the expression of brain aromatase; conversely, a lower temperature (20 degrees C) resulted in a decrease. ERalpha expression was depressed in accordance with the decrease of temperature, but ERbeta was unaffected by temperature. Between days 10 and 20, neither brain aromatase nor ERalpha expression was altered by temperature, whereas ERbeta expression was significantly enhanced by exposure to 32 degrees C. Between days 20 and 30, brain aromatase significantly increased at the higher temperature and decreased at 20 degrees C, but neither ERalpha nor ERbeta was affected by temperature. The expression of both brain aromatase and ERs, differentially regulated according to the temperature and to the developmental period, could be related to brain-sex differentiation. PMID- 12535178 TI - Evaluation of global analysis algorithms for single frequency fluorescence lifetime imaging microscopy data. AB - Global analysis of fluorescence lifetime image microscopy (FLIM) data can be used to obtain an accurate fit of multi-exponential fluorescence decays. In particular, it can be used to fit a bi-exponential decay to single frequency FLIM data, which is not possible with conventional fitting techniques. Bi-exponential fluorescence decay models can be used to analyse quantitatively single frequency FLIM data from samples that exhibit fluorescence resonance energy transfer (FRET). Global analysis algorithms simultaneously fit multiple measurements acquired under different experimental conditions to achieve higher accuracy. To demonstrate that bi-exponential models can indeed be fitted to single frequency data, we derive an analytical solution for the special case of two measurements and use this solution to illustrate the properties of global analysis algorithms. We also derive a novel global analysis algorithm that is optimized for single frequency FLIM data, and demonstrate that it is superior to earlier algorithms in terms of computational requirements. PMID- 12535179 TI - Transmitted light microscopy of a fibre reinforced metal. AB - A method is presented for studying fibre damage in continuous fibre reinforced composites. It is based on contrasting the transmission of light through intact translucent fibres with the light through fractured or dead-ended fibres. The method is applied in order to detect processing-induced fibre fractures in aluminium reinforced with continuous alumina fibres. PMID- 12535180 TI - Quantitative polarized light microscopy. AB - We describe a simple modification to a confocal microscope, which analyses the state of polarization of light emerging from the specimen so as to permit quantitative polarized light microscopy to be performed. The system uses a novel form of rotating analyser which, together with lock-in detection, permits images to be obtained where the image contrast corresponds to both specimen retardance and orientation (e.g. in the case of a birefringent specimen). Images are presented from a wide range of specimens and the origin of the contrast observed from simple point scatterers is investigated both theoretically and experimentally. PMID- 12535181 TI - Labelling quality and chromosome morphology after low temperature FISH analysed by scanning far-field and near-field optical microscopy. AB - A non-enzymatic, low temperature fluorescence in situ hybridization (LTFISH) procedure was applied to metaphase spreads and interphase cell nuclei. In this context 'low temperature' means that the denaturation procedure of the chromosomal target DNA usually applied by heat treatment and chaotropic agents such as formamide was completely omitted so that the complete hybridization reaction took place at 37 degrees C. For LTFISH, the DNA probe had to be single stranded, which was achieved by means of separate thermal denaturation of the DNA probe only. The DNA probe pUC1.77 was used for all LTFISH experiments. The labelling quality (number of binding sites, relative background intensity, relative intensity of major and minor binding sites) was analysed by confocal laser scanning microscopy (CLSM). An optimum in specificity and signal quality was obtained for 15 h hybridization time. For this hybridization condition of LTFISH, the chromosomal morphology was analysed by scanning near-field optical microscopy (SNOM). The results were compared with the morphology of chromosomes after (a) labelling of all centromeres using the same chemical treatment in the FISH procedure but with the application of target denaturation, and (b) labelling of all centromeres using a standard FISH protocol including thermal denaturation of the DNA probe and the chromosomal target. Depending on the FISH-procedure applied, SNOM images show substantial differences in the chromosome morphology. After LTFISH the chromosome morphology appeared to be much better preserved than after standard FISH. In contrast, the application of the LTFISH chemical treatment accompanied by heat denaturation had a very destructive influence on chromosomal morphology. The results indicate that, at least for certain DNA probes, specific chromosome labelling can be obtained without the usually applied heat and chemical denaturation of the DNA target, resulting in an apparently well preserved chromatin morphology as visualized by SNOM. LTFISH may be therefore a useful labelling technique whenever the chromosomal morphology had to be preserved after specific labelling of DNA regions. Binding mechanisms of single stranded DNA probes to double-stranded DNA targets are discussed. PMID- 12535182 TI - Automatic quantification of viability in epithelial cell cultures by texture analysis. AB - Quantification of live cells in phase contrast microscopy images allows in vivo assessment of the viability of cultured cells. An automatic screening procedure seems advisable because of the large number of cells that must be counted to achieve reasonable accuracy. This paper presents a method that quantifies necrosis in cell cultures by texture analysis of microscope images. The image is divided into regions of equal size that are classified by means of a segmentation algorithm based on texture analysis into three categories: live cells, necrotic cells and background. The classification uses three discriminant functions, built from parameters derived from the histogram and the co-occurrence matrix and calculated by performing an initial stepwise discriminant analysis on 21 sample images from a training set. The areas occupied by live and necrotic cells and number of live cells have been obtained for primary cellular cultures in intervals of 48 h during 2 weeks. The results have been compared with those obtained by an experienced observer, showing a very good correlation (Pearson's coefficient 0.95, kappa 0.87, N= 1600). A method has been developed that provides an accuracy similar to that provided by an expert, while allowing a much higher number of fields to be counted. PMID- 12535183 TI - A method of normalizing cathodoluminescence images of electron transparent foils for thickness contrast applied to InGaN quantum wells. AB - The uniformity of panchromatic cathodoluminescence (CL) from In0.09Ga0.91N/GaN quantum wells at 100 K was investigated using a combined transmission electron microscope-cathodoluminescence instrument. A technique for correcting CL images of electron-transparent wedge specimens for thickness contrast artefacts is presented. The foil thickness was estimated using the dynamical formulation of the relationship between the thickness and the (experimentally observed) transmitted electron intensity. For a given thickness the CL intensity was calculated using the Everhart-Hoff depth-dose function and also taking into account surface recombination losses. Experimental CL images were normalized by dividing by the calculated CL value at each point. The procedure was successful in calculating the underlying materials contrast in CL images of thin specimens of InGaN single quantum wells. Non-uniformities in the CL emission on the scale of approximately 0.7 micro m were observed. PMID- 12535184 TI - First tests of a dipole lens for a scanning electron microscope. AB - We have previously shown that a dipole lens has superior properties that are particularly suited for use in a low voltage scanning electron microscope (SEM) (Tsai & Crewe, 1996). The aberrations are lower than for any other type of lens and lead to a prediction of high resolution. We describe the construction details of a microscope based on this principle and present some early results. PMID- 12535185 TI - Fluorescence resonance energy transfer from cyan to yellow fluorescent protein detected by acceptor photobleaching using confocal microscopy and a single laser. AB - One manifestation of fluorescence resonance energy transfer (FRET) is an increase in donor fluorescence after photobleaching the acceptor. Published acceptor photobleaching methods for FRET have mainly used wide-field microscopy. A laser scanning confocal microscope enables faster and targeted bleaching within the field of view, thereby improving speed and accuracy. Here we demonstrate the approach with CFP and YFP, the most versatile fluorescent markers now available for FRET. CFP/YFP FRET imaging has been accomplished with a single laser (argon) available on virtually all laser-scanning confocal microscopes. Accordingly, we also describe the conditions that we developed for dual imaging of CFP and YFP with the 458 and 514 argon lines. We detect FRET in a CFP/YFP fusion and also between signalling molecules (TNF-Receptor-Associated-Factors or TRAFs) that are known to homo- and heterotrimerize. Importantly, we demonstrate that appropriate controls are essential to avoid false positives in FRET by acceptor photobleaching. We use two types of negative control: (a) an internal negative control (non-bleached areas of the cell) and (b) cells with donor in the absence of the acceptor (CFP only). We find that both types of negative control can yield false FRET. Given this false FRET background, we describe a method for distinguishing true positive signals. In summary, we extensively characterize a simple approach to FRET that should be adaptable to most laser-scanning confocal microscopes, and demonstrate its feasibility for detecting FRET between several CFP/YFP partners. PMID- 12535189 TI - In this issue: defined patient populations are required to establish genetic pathogenesis. PMID- 12535190 TI - WNTs: multiple genes, multiple functions. PMID- 12535191 TI - Homeobox genes, fetal wound healing, and skin regional specificity. PMID- 12535192 TI - Collagenase and impaired wound healing. PMID- 12535193 TI - Thrombospondin-1 plays a critical role in the induction of hair follicle involution and vascular regression during the catagen phase. AB - Hair growth is associated with pronounced vascular-endothelial-growth-factor induced perifollicular angiogenesis, whereas the catagen regression phase is characterized by apoptosis-driven blood vessel regression. The biologic relevance of endogenous inhibitors of angiogenesis in the control of hair cycling, however, has remained unknown. We studied the expression and biologic role of the angiogenesis inhibitor thrombospondin-1 (TSP-1) during the induced adult hair follicle cycle in wild-type, TSP-1 deficient, and TSP-1 overexpressing transgenic mice. TSP-1 expression was absent from hair bulb and dermal papilla cells during early to mid-anagen but was highly upregulated throughout the catagen involution phase. In TSP-1 deficient mice, the follicle growth phase was significantly prolonged, associated with increased perifollicular vascularization and vascular proliferation. Conversely, hair follicle growth was delayed in K14/TSP-1 transgenic mice that expressed high levels of TSP-1 in outer root sheath keratinocytes, associated with reduced perifollicular vascularization. These effects were most probably mediated via its antiangiogenic effects because TSP-1 did not affect the growth of cultured murine vibrissae in the absence of a functional vascular system. These results identify a critical role of TSP-1 in the induction of anagen follicle involution, with potential implications for the therapeutic modulation of hair follicle growth. PMID- 12535194 TI - Shift of localized growth zones contributes to skin appendage morphogenesis: role of the Wnt/beta-catenin pathway. AB - Skin appendage formation represents a process of regulated new growth. Bromodeoxyuridine labeling of developing chicken skin demonstrated the presence of localized growth zones, which first promote appendage formation and then move within each appendage to produce specific shapes. Initially, cells proliferate all over the presumptive skin. During the placode stage they are organized to form periodic rings. At the short feather bud stage, the localized growth zones shifted to the posterior and then the distal bud. During the long bud stage, the localized growth zones descended through the flank region toward the feather collar (equivalent to the hair matrix). During feather branch formation, the localized growth zones were positioned periodically in the basilar layer to enhance branching of barb ridges. Wnts were expressed in a dynamic fashion during feather morphogenesis that coincided with the shifting localized growth zones positions. The expression pattern of Wnt 6 was examined and compared with other members of the Wnt pathway. Early in feather development Wnt 6 expression overlapped with the location of the localized growth zones. Its function was tested through misexpression studies. Ectopic Wnt 6 expression produced abnormal localized outgrowths from the skin appendages at either the base, the shaft, or the tip of the developing feathers. Later in feather filament morphogenesis, several Wnt markers were expressed in regions undergoing rearrangements and differentiation of barb ridge keratinocytes. These data suggest that skin appendages are built to specific shapes by adding new cells from well-positioned and controlled localized growth zones and that Wnt activity is involved in regulating such localized growth zone activity. PMID- 12535195 TI - Fas and c-kit are involved in the control of hair follicle melanocyte apoptosis and migration in chemotherapy-induced hair loss. AB - Chemotherapy alters the structure and function of hair follicle melanocytes. Molecular mechanisms controlling melanocyte responses during chemotherapy-induced hair loss, however, remain largely unknown. Using immunohistology and multicolor confocal microscopy, we show here that cyclophosphamide administration to C57BL/6 mice alters the activity and fate of hair follicle melanocytes. After 24-48 h, hair bulb melanocytes expressing Fas undergo apoptosis. The number of apoptotic follicular melanocytes is significantly reduced (p<0.01) in cyclophosphamide treated Fas knockout mice compared to wild-type controls, suggesting that Fas signaling contributes to chemotherapy-induced melanocyte death. After 3-5 d, surviving hair bulb melanocytes express c-kit receptor, proliferate, and appear to migrate up the outer root sheath. Tyrosinase-positive and melanogenically active cells then appear in the epidermis. By Western blotting and immunohistochemistry, expression levels of the c-kit ligand, stem cell factor, in skin and epidermis are strongly increased after cyclophosphamide treatment. Cyclophosphamide-induced migration of the hair follicle melanocytes into epidermis is completely abrogated by administration of c-kit neutralizing antibody. These data suggest that chemotherapy induces a complex response in the hair follicle melanocytes, which includes apoptosis, proliferation, and migration. Pharmacologic manipulation of Fas and c-kit signaling pathways might be useful for the correction of skin hyperpigmentation as a side-effect of chemotherapy. PMID- 12535196 TI - Bone morphogenetic proteins and their antagonists in skin and hair follicle biology. AB - Bone morphogenetic proteins (BMP) are members of the transforming growth factor beta superfamily regulating a large variety of biologic responses in many different cells and tissues during embryonic development and postnatal life. BMP exert their biologic effects via binding to two types of serine/threonine kinase BMP receptors, activation of which leads to phosphorylation and translocation into the nucleus of intracellular signaling molecules, including Smad1, Smad5, and Smad8 ("canonical" BMP signaling pathway). BMP effects are also mediated by activation of the mitogen-activated protein (MAP) kinase pathway ("noncanonical" BMP Signaling pathway). BMP activity is regulated by diffusible BMP antagonists that prevent BMP interactions with BMP receptors thus modulating BMP effects in tissues. During skin development, BMPs its receptors and antagonists show stringent spatiotemporal expressions patterns to achieve proper regulation of cell proliferation and differentiation in the epidermis and in the hair follicle. In normal postnatal skin, BMP are involved in the control of epidermal homeostasis, hair follicle growth, and melanogenesis. Furthermore, BMP are implicated in a variety of pathobiologic processes in skin, including wound healing, psoriasis, and carcinogenesis. Therefore, BMPs represent new important players in the molecular network regulating homeostasis in normal and diseased skin. Pharmacologic modulation of BMP signaling may be used as a new approach for managing skin and hair disorders. PMID- 12535197 TI - Apoptosis regulators and responses in human melanocytic and keratinocytic cells. AB - Apoptosis in keratinocytes is required for epidermal turnover, stratum corneum formation, and removal of ultraviolet-damaged premalignant cells. Its role in melanocyte homeostasis and transformation, on the other hand, has not been defined, although apoptosis resistance is a commonly recognized feature of melanoma. We examined the expression of apoptosis regulators in melanocytes, keratinocytes, melanoma, and HaCat cells. Melanocytic cells expressed relatively high levels of Bcl-2, Bcl-X(L), Mcl-1, C-IAP-1, C-IAP-2, XIAP, Livin, and Apaf-1. The only apoptotic regulator that was differentially expressed in melanoma cells and not melanocytes was Survivin, whereas Bax was expressed in melanocytes but not in most melanoma lines. Keratinocytic cells, on the other hand, expressed high levels of FLIP and were relatively deficient in Bcl-2 family proteins. Levels of p53 were highest in HaCat cells and some of the melanoma lines, and barely detectable in melanocytes and keratinocytes. Next, susceptibility of these cells types to apoptosis induced by ultraviolet B, the tyrosine analog 4-tert butylphenol, and cytotoxic drugs was examined. Melanocytes were relatively resistant to ultraviolet B, whereas keratinocytes were unresponsive to 4-tert butylphenol. Melanocytes and keratinocytes were generally less susceptible than melanoma lines and HaCat cells to etoposide, cisplatin, and staurosporine. Induction of apoptosis in these cell types was generally associated with decreased levels of Mcl-1, XIAP, and Livin, and increased levels of p53, whereas levels of other apoptotic regulators were unaltered. These results provide insights into the potential roles of apoptosis in the function and transformation of epidermal melanocytes and keratinocytes. PMID- 12535198 TI - Helium-neon laser irradiation stimulates migration and proliferation in melanocytes and induces repigmentation in segmental-type vitiligo. AB - Low-energy helium-neon lasers (632.8 nm) have been employed in a variety of clinical treatments including vitiligo management. Light-mediated reaction to low energy laser irradiation is referred to as biostimulation rather than a thermal effect. This study sought to determine the theoretical basis and clinical evidence for the effectiveness of helium-neon lasers in treating vitiligo. Cultured keratinocytes and fibroblasts were irradiated with 0.5-1.5 J per cm2 helium-neon laser radiation. The effects of the helium-neon laser on melanocyte growth and proliferation were investigated. The results of this in vitro study revealed a significant increase in basic fibroblast growth factor release from both keratinocytes and fibroblasts and a significant increase in nerve growth factor release from keratinocytes. Medium from helium-neon laser irradiated keratinocytes stimulated [3H]thymidine uptake and proliferation of cultured melanocytes. Furthermore, melanocyte migration was enhanced either directly by helium-neon laser irradiation or indirectly by the medium derived from helium neon laser treated keratinocytes. Thirty patients with segmental-type vitiligo on the head and/or neck were enrolled in this study. Helium-neon laser light was administered locally at 3.0 J per cm2 with point stimulation once or twice weekly. The percentage of repigmented area was used for clinical evaluation of effectiveness. After an average of 16 treatment sessions, initial repigmentation was noticed. Marked repigmentation (>50%) was observed in 60% of patients with successive treatments. Basic fibroblast growth factor is a putative melanocyte growth factor, whereas nerve growth factor is a paracrine factor for melanocyte survival in the skin. Both nerve growth factor and basic fibroblast growth factor stimulate melanocyte migration. It is reasonable to propose that helium-neon laser irradiation clearly stimulates melanocyte migration and proliferation and mitogen release for melanocyte growth and may also rescue damaged melanocytes, therefore providing a microenvironment for inducing repigmentation in vitiligo. PMID- 12535199 TI - A commercial sunscreen's protection against ultraviolet radiation-induced immunosuppression is more than 50% lower than protection against sunburn in humans. AB - Ultraviolet radiation (280-400 nm)-induced suppression of cutaneous cell-mediated immunity plays an important part in the development of skin cancer. Sunscreens are widely advocated to protect against skin cancer but if they offer insufficient protection against immunosuppression they may inadvertently increase skin cancer risk. This human study evaluated immunoprotection afforded by a commercial sunscreen preparation (labeled sun protection factor 15) offering primarily ultraviolet B (280-320 nm) protection. Indirectly, it also investigated whether ultraviolet A (320-400 nm) plays a part in ultraviolet radiation-induced immunosuppression. Healthy white-skinned volunteers were used (n=119). Ultraviolet radiation exposures were on previously unexposed buttock skin with an ultraviolet radiation source that complied with European recommendations for sunscreen testing. Ultraviolet radiation dose-response curves for sunburn/erythema and suppression of the contact hypersensitivity response were generated either with or without sunscreen in vivo and protection factors were derived for both end-points. The ultraviolet radiation wavelengths transmitted by the sunscreen were determined in vitro and showed that the sunscreen was primarily an ultraviolet B absorber, with relatively poor absorption in the ultraviolet A region. The sun-screen protected against both erythema and immunosuppression but protection against immunosuppression (IPF=4.9, 95% confidence interval: 2.3-10.6) was less than half that for erythema (Ery PFg=14.2, 95% confidence interval: 10.2-19.8). Failure of the sunscreen to afford comparable protection against both end-points was probably due to immunosuppression by ultraviolet A, a part of the solar spectrum that does not readily cause sunburn. The sunscreen protected against both end-points, which supports the use of sunscreens to reduce immunosuppression but protection against immunosuppression may be improved if sunscreens are formulated to offer equivalent protection against ultraviolet B and ultraviolet A. PMID- 12535200 TI - Terahertz pulse imaging of ex vivo basal cell carcinoma. AB - Terahertz pulse imaging has been used for the first time to study basal cell carcinoma ex vivo, the most common form of skin cancer. This noninvasive technique uses part of the electromagnetic spectrum in the frequency range 0.1 2.7 THz. A total of 21 samples were imaged; the study was performed blind and results were compared to histology. Each image consisted of possible diseased tissue and normal tissue from the same patient. The diseased tissue showed an increase in absorption compared to normal tissue, which is attributed to either an increase in the interstitial water within the diseased tissue or a change in the vibrational modes of water molecules with other functional groups. Seventeen of the images showed a significant difference between the normal and the diseased tissue. These were confirmed by histology to be basal cell carcinomas. Of the remaining four cases, three showed no contrast and were confirmed as blind controls of normal tissue; the fourth case was a suspected basal cell carcinoma but showed no contrast, and histology showed no tumor. Cross-sections of the terahertz images, showing the terahertz absorption, were compared to histology. Regions of increased terahertz absorption agreed well with the location of the tumor sites. Resolutions at 1 THz of 350 microm laterally and 40 microm axially in skin were attainable with our system. These results demonstrate the ability of terahertz pulse imaging to distinguish basal cell carcinoma from normal tissue, and this macroscopic technique may, in the future, help plan surgery. PMID- 12535201 TI - Characteristics of the epidermis and stratum corneum of hairless mice with experimentally induced diabetes mellitus. AB - Diabetes mellitus induces many pathophysiologic changes in the skin. Even so, dermatologists still lack an animal model of diabetes that enables the direct evaluation of the various functional properties of the skin. Our group induced two types of an experimental type 1 diabetes model in hairless mice by administering either streptozotocin or alloxan, in order to examine the properties of the stratum corneum and epidermis of these animals. The plasma glucose concentrations of the mice at 3 wk after their i.v. injection were significantly higher than those of control mice (streptozotocin, 3.2-fold; alloxan, 3.7-fold). The stratum corneum water content was significantly reduced in both types of diabetic mice, whereas the transepidermal water loss remained unchanged. The amino acid content with normal epidermal profilaggrin processing was either normal or elevated in the stratum corneum of the streptozotocin treated mice. In contrast, the stratum corneum triglyceride content in the streptozotocin-treated mice was significantly lower than the control level, even though the levels of ceramides, cholesterols, and fatty acids in the stratum corneum were all higher than the control levels. The streptozotocin-treated group also exhibited decreases in basal cell proliferation and epidermal DNA content linked with an increase in the number of corneocyte layers in the stratum corneum, suggesting that the rates of epidermal and stratum corneum turnover were slower in the streptozotocin-treated animals than in the normal controls. In contrast, there were no remarkable changes in any of the epidermal differentiation marker proteins examined. This finding in diabetic mice, namely, reduction in both the epidermal proliferation and stratum corneum water content without any accompanying impairment in the stratum corneum barrier function, is similar to that found in aged human skin. Our new animal model of diabetes will be useful for the study of diabetic dermopathy as well as the mechanisms of stratum corneum moisturization. PMID- 12535202 TI - Pertussis toxin-sensitive secretory phospholipase A2 expression and motility in activated primary human keratinocytes. AB - Secretory phospholipase A2 and cycloxygenase-2 are coexpressed in activated primary keratinocytes. These proteins are known to be functionally linked, mediating proliferation of human keratinocytes during epidermal wound repair. Primary human keratinocytes grown at low densities (15-30%; nonconfluent) produce high levels of prostaglandin E2 important for proliferation and are a good model for studying activated keratinocytes after injury. In this study, we used this model to assess the role of secretory phospholipase A2 and cycloxygenase-2 in keratinocyte motility. Initial work showed 24 h pretreatment with 20 ng pertussis toxin per ml, an inhibitor of the inhibitory G-protein, decreased prostaglandin E2 production and both secretory phospholipase A2 and cycloxygenase-2 protein expression. This suggested that inhibitory G-protein may be involved in mediating expression of these proteins. Pertussis toxin also caused changes in cell morphology, actin organization, and keratinocyte motility. Pretreatment with 5 microm 12-epi-scalaradial, a secretory phospholipase A2 inhibitor, caused similar changes in cell motility and actin organization; however, the specific cycloxygenase-2 inhibitor, SC-58236 (20 nm) was much less effective. These results suggested that secretory phospholipase A2 plays a part in keratinocyte motility that is independent of its functional linkage to cycloxygenase-2 and prostaglandin E2 biosynthesis. PMID- 12535203 TI - Identification of recurrent mutations in the ARS (component B) gene encoding SLURP-1 in two families with mal de Meleda. AB - Mal de Meleda is a rare, autosomal recessive form of palmoplantar keratoderma. The disease has been mapped to chromosome 8qter, and in a recent study mutations in the ARS gene have been identified in families with this disorder. Here, we report two unrelated families with mal de Meleda, in which two different homozygous mutations in the ARS gene were identified. These findings support the notion that mutations in the ARS gene are pathogenic in mal de Meleda. PMID- 12535204 TI - Interleukin-10 promoter polymorphisms and susceptibility to skin squamous cell carcinoma after renal transplantation. AB - After organ transplantation, susceptibility to cancer is multifactorial, especially for skin carcinomas. Risk factors may include genetic susceptibilities, such as the control of cytokine production. Interleukin-10 is a cytokine that is implicated in tumorigenesis, and it has been shown that polymorphisms in its gene promoter correlate with differential amounts of production. The aim of this study was to investigate a possible association between interleukin-10 gene promoter polymorphisms and the occurrence of skin carcinomas after renal transplantation. Seventy kidney transplant recipients who developed a squamous cell carcinoma or a basal cell carcinoma were examined for polymorphisms in the interleukin-10 gene promoter using polymerase chain reaction based methods. Single base pair mutations were studied at positions -1082, -819, and -592. These patients were compared to 70 healthy controls and to 70 matched renal transplant recipients without cancer. The interleukin-10 secretion capability was tested in a subgroup of 40 of these patients by in vitro stimulation of peripheral mononuclear cells. Interleukin-10 genotypes and haplotypes were differently distributed in kidney transplant recipients who developed a skin carcinoma, but especially a squamous cell carcinoma, with an increased frequency of the GCC haplotype and a decreased frequency of the ATA haplotype. Subsequently, we found a shift in the predicted phenotypes from the low production phenotype to the high production phenotype. Secretion of interleukin-10 was strongly correlated to the production predicted phenotype, and tended to be higher in patients who developed a squamous cell carcinoma than in the others. These results indicate that interleukin-10 gene polymorphisms and interleukin-10 production capability may contribute to the development of skin squamous cell carcinomas after renal transplantation. PMID- 12535205 TI - The role of subclass switching in the pathogenesis of endemic pemphigus foliaceus. AB - Endemic pemphigus foliaceus, like the sporadic form seen in the developed world, is mediated by IgG antibodies to desmoglein-1. We studied an endemic focus in Limao Verde, Brazil, where disease prevalence is 3.4%. We previously detected IgG antibodies to desmoglein-1 in 97% of patients, but also in 55% of normal subjects in the endemic focus, with progressively lower levels in normal subjects in surrounding areas. An environmental trigger is hypothesized to explain these and other findings. In this study we sought to determine if patients and enzyme linked-immunosorbent-assay-positive normal subjects in Limao Verde differ in IgG subclass response to desmoglein-1. We developed a sensitive and specific subclass enzyme-linked immunosorbent assay using recombinant desmoglein-1 and standardized the assay to enable comparability between the four subclasses. We found that normal subjects have an IgG1 and IgG4 response, whereas patients have similar levels of IgG1 but a mean 19.3-fold higher IgG4 response. Patients in remission have a weak IgG4 response, and a 74.3-fold higher IgG4 response is associated with active disease. Finally, in five patients in whom we had blood samples from both before and after the onset of clinical disease, a mean 103.08-fold rise in IgG4 was associated with onset of clinical disease, but only a mean 3.45-fold rise in IgG1. These results suggest that the early antibody response in normal subjects living in the endemic area and in patients before the onset of clinical disease is mainly IgG1. Acquisition of an IgG4 response is a key step in the development of clinical disease. PMID- 12535206 TI - Farnesyl thiosalicylic acid chemosensitizes human melanoma in vivo. AB - Malignant melanoma is well known for its poor response to a variety of chemotherapeutic agents. Testing of numerous treatment strategies has identified dacarbazine as the most active single drug; however, its response rates in various clinical settings are quite limited. Defective apoptosis in combination with oncogenic proteins (such as activated Ras) in cell proliferation pathways plays a key part in both the development and disease progression of human melanoma. Farnesyl thiosalicylic acid, a novel Ras inhibitor, dislodges Ras proteins from the cell membrane, leading to inhibition of cell transformation and tumor growth. In this study we evaluated the effect of farnesyl thiosalicylic acid treatment on established human melanoma xenografts grown in mice with severe combined immunodeficiency as well as the chemosensitizing effect of farnesyl thiosalicylic acid in combination with dacarbazine. Daily administration of 10, 20, or 40 mg per kg of farnesyl thiosalicylic acid resulted in a concentration dependent reduction in tumor growth, with growth inhibition reaching a mean value of 45+/-7%, at the highest concentration. The combination of farnesyl thiosalicylic acid (10 mg per kg per day) and dacarbazine (80 mg per kg per day) resulted in a significant reduction of 56%+/-9%, in mean tumor growth. Analysis of toxicologic parameters (mouse weight, blood cell counts, and blood chemistry) showed an acceptable and similar toxicity profile for both the single-agent farnesyl thiosalicylic acid treatment and the combination of farnesyl thiosalicylic acid plus dacarbazine treatment. Given the observed preclinical treatment responses and the low toxicity, our results support the notion that farnesyl thiosalicylic acid in combination with dacarbazine may qualify as a rational treatment approach for human melanoma. PMID- 12535207 TI - IL-12 completely blocks ultraviolet-induced secretion of tumor necrosis factor alpha from cultured skin fibroblasts and keratinocytes. AB - Interleukin-12 is an important regulator of other cytokines. Although interleukin 12 is considered to act primarily on lymphocytes, provoking a shift from T helper 2 to T helper 1 cells and an increase in lymphocyte-derived tumor necrosis factor alpha, we hypothesized that interleukin-12 might also affect tumor necrosis factor alpha secretion from skin cells. In this study, keratinocytes were treated with ultraviolet-B, ultraviolet-A, or sham irradiation, without or with exogenous interleukin-12. Remarkably, the exogenous interleukin-12 totally blocked ultraviolet-B-induced tumor necrosis factor alpha production. Both ultraviolet-A and ultraviolet-B were capable of inducing interleukin-12 production. To determine the molecular mechanism of this effect, we used a chloramphenicol acetyl transferase reporter under the control of a 1.2 kb fragment of the wild type (-308G) human tumor necrosis factor alpha promoter and found significant suppression of promoter activity with interleukin-12. Studies using the -308A variant of the human tumor necrosis factor alpha promoter showed much higher promoter activity overall, but also a greater sensitivity to suppression by interleukin-12. The mechanism did not involve blockage of the interleukin-1 receptor, because interleukin-12 did not suppress interleukin-1-mediated induction of collagenase mRNA. To determine the role of endogenous interleukin 12, we found that anti-interleukin-12 antibodies enhanced ultraviolet-B-induced tumor necrosis factor alpha secretion. Thus, interleukin-12 strongly inhibits tumor necrosis factor alpha production by noninflammatory skin cells, mostly or entirely through inhibition of gene transcription via an element within the first 1.2 kb of the tumor necrosis factor alpha promoter. The result is a shift in tumor necrosis factor alpha production from noninflammatory cells to T helper 1 cells. Because tumor necrosis factor alpha is central to the pathogenesis of several photosensitive skin diseases and certain forms of immune suppression, interleukin-12 may have important physiologic, pathophysiologic, and therapeutic roles. PMID- 12535208 TI - Ultraviolet B irradiation of the eye activates a nitric oxide-dependent hypothalamopituitary proopiomelanocortin pathway and modulates functions of alpha melanocyte-stimulating hormone-responsive cells. AB - Ultraviolet B radiation increases DOPA-positive melanocytes in the skin specifically at the site of exposure. We found unexpectedly that ultraviolet B irradiation of the eye increased the concentration of alpha-melanocyte stimulating hormone in plasma and systemically stimulated epidermal melanocytes in mice. To test the possible involvement of hypothalamopituitary proopiomelanocortin system in the systemic activation of skin melanocytes, ultraviolet B was also irradiated to the eye after hypophysectomy. Hypophysectomy strongly inhibited the ultraviolet B-induced stimulation of melanocytes. To elucidate the pathway by which ultraviolet B irradiation of the eye activated the hypothalamopituitary system, we examined the effect of bilateral ciliary ganglionectomy and denervation of the optic nerves on the ultraviolet B-induced melanocyte stimulation. Ciliary ganglionectomy, but not optic nerve denervation, strongly inhibited melanocyte stimulation by localized irradiation of the eye. Furthermore, melanocyte stimulation by localized ultraviolet B irradiation of the eye was not observed in mice that lack the inducible type of nitric oxide synthase. These results clearly indicate that a signal evoked by ultraviolet B irradiation of the eye is transmitted in a nitric oxide-dependent manner through the ciliary ganglia involving the first branch of the trigeminal nerve to the hypothalamopituitary proopiomelanocortin system, resulting in upregulation of alpha-melanocyte-stimulating hormone secretion and consequent stimulation of melanocytes in the skin. The novel network involving the trigeminal nerve and nitric oxide-dependent signaling pathway might play important parts in the activation of proopiomelanocortin-dependent biologic reactions, such as alpha melanocyte-stimulating hormone-induced stimulation of melanocytes in the skin, in ultraviolet B-enriched environments. PMID- 12535209 TI - Possible involvement of gelatinases in basement membrane damage and wrinkle formation in chronically ultraviolet B-exposed hairless mouse. AB - A number of studies indicate that matrix metalloproteinase might be involved in photoaging, but little is known about their direct contribution to ultraviolet induced histologic and morphologic changes in the skin in vivo. This study reports the relationship between changes of matrix metalloproteinase activities and ultraviolet B-induced skin changes in hairless mouse. The role of matrix metalloproteinase in the skin changes was studied by topical application of a specific matrix metalloproteinase inhibitor. The backs of mice were exposed to ultraviolet B three times a week for 10 wk. Histologic studies showed that the basement membrane structure was damaged, with epidermal hyperplasia, in the first 2 wk of ultraviolet B irradiation, followed by the appearance of wrinkles, which gradually extended in the latter half of the ultraviolet B irradiation period. We observed enhancement of type IV collagen degradation activity, but not collagenase or matrix metalloproteinase-3 activity, in extracts of ultraviolet B irradiated, wrinkle-bearing skin. Gelatin zymographic analysis revealed that gelatinases, matrix metalloproteinase-9 and matrix metalloproteinase-2, were significantly increased in the extract. In situ zymographic study clarified that the activity was specifically localized in whole epidermis of ultraviolet B irradiated, wrinkled skin in comparison with normal skin. The activity was induced around the basal layer of the epidermis by a single ultraviolet exposure of at least one minimal erythema dose. Furthermore, topical application of a specific matrix metalloproteinase inhibitor, CGS27023A, inhibited ultraviolet B induced gelatinase activity in the epidermis, and its repeated application prevented ultraviolet B-induced damage to the basement membrane, as well as epidermal hyperplasia and dermal collagen degradation. Ultraviolet B-induced wrinkles were also prevented by administration of the inhibitor. These results, taken together, suggest that ultraviolet B-induced enhancement of gelatinase activity in the skin contributes to wrinkle formation through the destruction of basement membrane structure and dermal collagen in chronically ultraviolet B exposed hairless mouse, and thus topical application of matrix metalloproteinase inhibitors may be an effective way to prevent ultraviolet B-induced wrinkle formation. PMID- 12535210 TI - Deletion of the homeobox gene PRX-2 affects fetal but not adult fibroblast wound healing responses. AB - The phenotype of fibroblasts repopulating experimental wounds in vivo has been shown to influence both wound healing responses and clinical outcome. Recent studies have demonstrated that the human homeobox gene PRX-2 is strongly upregulated in fibroblasts within fetal, but not adult, mesenchymal tissues during healing. Differential homeobox gene expression by fibroblasts may therefore be important in mediating the scarless healing exhibited in early fetal wounds. RNase protection analysis demonstrated that murine Prx-2 expression was involved in fetal but not adult wound healing responses in vitro. Using fibroblasts established from homozygous mutant (Prx-2-/-) and wild-type (Prx 2+/+) murine skin tissues it was demonstrated that Prx-2 affected a number of fetal fibroblastic responses believed to be important in mediating scarless healing in vivo; namely cellular proliferation, extracellular matrix reorganization, and matrix metalloproteinase 2 and hyaluronic acid production. These data demonstrate how Prx-2 may contribute to the regulation of fetal, but not adult, fibroblasts and ultimately the wound healing phenotype. This study provides further evidence for the importance of homeobox transcription factors in the regulation of scarless wound healing. A further understanding of these processes will, it is hoped, enable the targeting of specific therapies in wound healing, both to effect scarless healing and to stimulate healing in chronic, nonhealing wounds such as venous leg ulcers. PMID- 12535211 TI - Different expression of mu-opiate receptor in chronic and acute wounds and the effect of beta-endorphin on transforming growth factor beta type II receptor and cytokeratin 16 expression. AB - There is evidence that neuropeptides, especially the opiate receptor agonists, are involved in wound healing. We have previously observed that beta-endorphin, the endogenous ligand for the mu-opiate receptor, stimulates the expression of cytokeratin 16 in a dose-dependent manner in human skin organ cultures. Cytokeratin 16 is expressed in hyperproliferative epidermis such as psoriasis and wound healing. Therefore we were interested to study whether epidermal mu-opiate receptor expression is changed at the wound margins in acute and chronic wounds. Using classical and confocal microscopy, we were able to compare the expression level of mu-opiate receptors and the influence of beta-endorphin on transforming growth factor beta type II receptor in organ culture. Our results show indeed a significantly decreased expression of mu-opiate receptors on keratinocytes close to the wound margin of chronic wounds compared to acute wounds. Additionally beta endorphin upregulates the expression of transforming growth factor beta type II receptor in human skin organ cultures. These results suggest a crucial role of opioid peptides not only in pain control but also in wound healing. Opioid peptides have already been used in animal models in treatment of wounds; they induce fibroblast proliferation and growth of capillaries, and accelerate the maturation of granulation tissue and the epithelization of the defect. Furthermore opioid peptides may fine-tune pain and the inflammatory response while healing takes place. This new knowledge could potentially be used to design new locally applied drugs to improve the healing of painful chronic wounds. PMID- 12535212 TI - Severely impaired wound healing in the collagenase-resistant mouse. AB - Collagen in the skin undergoes dramatic reorganization during wound repair. Matrix metalloproteinases degrade and remodel the collagen in a tightly controlled process. The collagenase-resistant mouse, Col1a1(tm1Jae), has been developed to produce collagen type I, which is resistant to degradation by human matrix metalloproteinase 1. These mice grow normally but develop thickened skin with age. We investigated the effect of this mutant collagen on wound repair. Incisional wounds were made on Col1a1(tm1Jae) homozygous mutant (Col1a1(r/r)) and wild-type (Col1a1+/+) mice and these wounds were harvested at 1 and 6 h, 1, 2, 3, 7, 10, 14, and 70 d post wounding. Wound healing was severely delayed in Col1a1(r/r) wounds, with wounds remaining significantly wider than wild-type for the first 2 wk after injury. Reepithelialization of the Col1a1(r/r) wounds took 7 d longer than in the wild-type. The Col1a1(r/r) wounds had a prolonged early inflammatory response. Immunostaining for matrix metalloproteinases revealed significant upregulation of matrix metalloproteinase 13 in Col1a1(r/r) wounds, but minimal changes in other matrix metalloproteinases. There was no significant difference in scarring between Col1a1(r/r) and Col1a1+/+ wounds after 70 d. PMID- 12535213 TI - Fas ligand in pemphigus sera induces keratinocyte apoptosis through the activation of caspase-8. AB - The Fas/Fas ligand system triggers the extrinsic apoptotic pathway and is involved in several inflammatory conditions, also at the skin level. The Fas/Fas ligand cell death pathway plays a major role in anoikis, a type of apoptosis characterized by cell detachment. As pemphigus is characterized by loss of cell to cell adhesion, we evaluated the role of anoikis and Fas ligand in this bullous disease. We report that, in suprabasal epidermis from perilesional pemphigus skin, most keratinocytes are apoptotic. Moreover, Fas ligand levels are markedly increased in sera from pemphigus patients, whereas they are undetectable in sera from patients undergoing steroid treatment. Sera from untreated patients but not from patients under steroids induce keratinocyte apoptosis. Pemphigus-sera induced cell death is partially inhibited by pretreatment with anti-Fas ligand antibodies and by incubation with caspase-8 inhibitor Z-IETD-FMK. Finally, caspase-8 is activated in keratinocytes provided with sera from pemphigus patients, whereas cleavage is partially blocked by pretreatment of sera with anti Fas ligand antibody. These results suggest that increased Fas ligand in pemphigus sera is responsible for keratinocyte apoptosis, which occurs through the activation of a caspase-8-driven extrinsic apoptotic pathway. PMID- 12535214 TI - p75 Neurotrophin receptor antagonist retards apoptosis-driven hair follicle involution (catagen). PMID- 12535215 TI - Novel transglutaminase 1 gene mutations (R348X/Y365D) in a Japanese family with lamellar ichthyosis. PMID- 12535219 TI - The cellular rejection of xenografts--recent insights. PMID- 12535220 TI - Presidential address: 6th Congress of the International Xenotransplantation Association. PMID- 12535221 TI - Experimental xenotransplantation: a personal history. PMID- 12535222 TI - Are anti-endothelial cell antibodies a pre-requisite for the acute vascular rejection of xenografts? AB - BACKGROUND: Vascular rejection occurring within the first few weeks after transplantation is still the major immunological barrier to the long term survival of xenografts. Currently there is no consensus about what to call this type of rejection (acute vascular rejection, delayed xenograft rejection or acute humoral xenograft rejection), nor about how to prevent or treat it. METHODS: A review of published evidence to define the heterogeneity of this phase of rejection and examine the role of antibodies, complement and graft-infiltrating inflammatory cells. RESULTS: i) antibodies are always involved in acute vascular rejection; ii) this antibody-mediated rejection may be complement-dependent or independent; iii) inflammatory cells may mediate an antibody- and complement independent phase of rejection in some small animal models (which, in its pure form cannot be called 'vascular rejection') iv) there remain significant questions about the relevance of 'accommodation' and the importance of coagulation abnormalities. CONCLUSIONS: Without doubt, future research would be helped by distinguishing between these different forms of delayed xenograft rejection, using terminology to reflect the involvement of specific pathophysiological mechanisms. An updated classification of the stages of xenograft rejection is proposed here. PMID- 12535223 TI - Fetal porcine thymus engraftment, survival and CD4 reconstitution in alphaGal-KO mice is impaired in the presence of high levels of antibodies against alphaGal. AB - Xenospecific T-cell tolerance can be induced among murine and human T-cells by porcine thymic grafting. However, anti-alpha 1,3-galactosyltranserase (alphaGal) (Galalpha1-3Galbeta1-4GlcNAc-R) natural antibodies (NAbs) pose a major barrier to porcine xenografts in humans. We used alphaGal knockout (KO) and muchain KO mice to explore the effect of natural anti-alphaGal and other xenoantibodies on porcine thymic engraftment and to examine the potential of thymic tissue to tolerize anti-alphaGal antibody-producing cells. Thymectomized [adult thymectomy (ATX)] non-immunized and rabbit red blood cell (RRBC) pre-transplant immunized alphaGal-KO (knockout), wild-type (WT) and mu chain KO B6 mice were treated with 3Gy total body irradiation (TBI), and T and natural killer (NK) cell depleting monoclonal antibodies (mAbs). These conditioned mice were grafted with fetal porcine thymus and liver (FP THY/LIV) tissue under the kidney capsule. Flow cytometric analysis was performed to follow CD4 reconstitution as a measure of FP THY engraftment and function. Only mice with >10% CD4+ peripheral blood lymphocytes (PBL) were considered successfully engrafted. Enzyme-linked immunosorbent assay (ELISA) was used to assess the kinetics of immunoglobulin M (IgM) and IgG anti-alphaGal antibodies. Anti-pig antibodies were monitored by flow cytometry (FCM). FP THY engrafted successfully in most of the immunoglobulin deficient mice (11 out of 12, 92%) and the outcome was similar in WT B6 controls (8 out of 12, 67%). Non-immunized alphaGal-KO mice grafted with FP THY had a similar success rate (7 out of 11) to that observed in non-immunized alphaGal-WT controls (2 out of 4). In contrast, alphaGal-KO mice immunized pre-transplant with RRBC, then grafted with FP THY/LIV, showed a significant reduction in the success of thymic grafting (2 out of 9, 22%) compared with pre-transplant immunized WT controls (4 out of 7; 57%) and non-immunized alphaGal-KO mice (7 out of 11, 64%). Anti-Gal and anti-pig antibody levels were not markedly augmented by porcine thymus grafts in mice with successful thymus grafts. FP THY engraftment is impaired in the presence of high levels of anti-alphaGal xenoantibodies. However, low levels of anti-alphaGal antibodies and other mouse anti-pig NAbs appear not to play a major role in the rejection of FP THY. Although grafting FP THY expressing the alphaGal epitope did not tolerize B cells producing anti alphaGal antibodies in a T-cell independent manner, it prevented T-cell dependent sensitization by inducing T-cell tolerance to porcine antigens. PMID- 12535224 TI - Immune parameters relevant to neural xenograft survival in the primate brain. AB - The lack of supply and access to human tissue has prompted the development of xenotransplantation as a potential clinical modality for neural cell transplantation. The goal of the present study was to achieve a better understanding of the immune factors involved in neural xenograft rejection in primates. Initially, we quantified complement mediated cell lysis of porcine fetal neurons by primate serum and demonstrated that anti-C5 antibody treatment inhibited cell death. We then developed an immunosuppression protocol that included in vivo anti-C5 monoclonal antibody treatment, triple drug therapy (cyclosporine, methylprednisolone, azathioprine) and donor tissue derived from CD59 or H-transferase transgenic pigs and applied it to pig-to-primate neural cell transplant models. Pre-formed alphaGal, induced alphaGal and primate anti mouse antibody (PAMA) titers were monitored to assess the immune response. Four primates were transplanted. The three CD59 neural cell recipients showed an induced anti-alphaGal response, whereas the H-transferase neural cell recipient exhibited consistently low anti-alphaGal titers. Two of these recipients contained surviving grafts as detected by immunohistochemistry using selected neural markers. Graft survival correlated with high dose cyclosporine treatment, complete complement blockade and the absence of an induced PAMA response to the murine anti-C5 monoclonal antibodies. PMID- 12535225 TI - Exploiting virus stealth technology for xenotransplantation: reduced human T cell responses to porcine cells expressing herpes simplex virus ICP47. AB - Direct recognition of porcine major histocompatibility complex (MHC) proteins by human T cells is well documented. Eliminating donor (porcine) MHC proteins may therefore be beneficial in pig-to-human xenotransplants. To this end, we have attempted to exploit viral stealth mechanisms to eliminate pig MHC class I cell surface expression. PK(15) (pig kidney) cells stably transfected with the herpes simplex virus (HSV) ICP47 gene [PK(15)-ICP47 cells] exhibited a dramatic reduction of MHC class I cell-surface expression when compared with untransfected PK(15) cells. To test the effect of down-regulation of porcine MHC class I on human cellular immune responses, a human CD8+ enriched T cell line (anti-PK15 T cells) with reactivity towards PK(15) cells was derived by repeated stimulation of human T cells with PK(15) cells stably transfected with the costimulatory molecule B7.1 [PK(15)-B7.1 cells]. Anti-PK15 T cells efficiently lyzed PK(15) cells but not PK(15)-ICP47 (class I negative) cells. Consistent with effector function, anti-PK15 T cells showed a robust proliferative response to PK(15)-B7.1 cells but did not proliferate at all to PK(15)-B7.1 cells which also expressed HSV ICP47. These results suggest that virus stealth technology can be exploited for xenotransplantation. PMID- 12535226 TI - Xenogeneic milieu markedly remodels endocrine cell populations after transplantation of fish islets into streptozotocin-diabetic nude mice. AB - It is unknown whether irrelevant foreign endocrine products secreted by xenografts would be biologically active and potentially harmful to recipients; even if entirely inert, continuous production might result in harmful circulating antigen-antibody complexes. We examined the fate of such a product using a fish (tilapia)-to-mouse islet xenograft model. Teleost fish islets, like mammalian islets, are composed primarily of cells producing insulin, glucagon or somatostatin; however, teleost fish have two different populations of somatostatin (SST) producing delta cells, one producing SST-14, a 14 amino acid SST identical to mammalian SST, which is derived from the pre-proSST-I gene which is present in all vertebrates, and the other a "large" (i.e. 22 to 28 amino acid) SST derived from a pre-proSST-II gene, which is not found in mammals. In contrast to 'large' SST, which has no mammalian homolog, teleost fish insulins, glucagons and SST-14 exhibit significant biological activity in mammals. Tilapia islets were transplanted under the kidney capsules of streptozotocin-diabetic nude mice, and mice with functioning grafts were killed at various times after transplantation. Serial sections of graft-bearing kidneys were stained by immunoperoxidase for insulin, SST-14, SST-25 and glucagon positive cells, and the areas of each cell type in the graft were measured using image analysis. Sections of untransplanted tilapia islets (both in situ and after harvest/culture) were also immunostained and measured as controls. Xenotransplantation of fish islets into diabetic nude mice resulted in the rapid degeneration and near total loss of SST-25+ cells, as well as a marked redistribution of the proportions of the remaining endocrine cell types. The proportions of cell types in the grafts gradually changed from a piscine pattern to that of mammalian islets. PMID- 12535227 TI - Pre-transplant analysis of accommodation in donor pigs. AB - Accommodation could lead to xenograft acceptance without the need for severe immune suppression. Generally graft accommodation is appreciated in the sensitized recipient, after transplantation. By inducing accommodation in chimeric donors, however, the risk and cost of inducing accommodation in the recipient would be reduced. An indirect assay of accommodation in the donor pig is needed for screening donors prior to procurement of the xenograft. The resistance of peripheral blood lymphocytes to cytolysis by antibody and complement was assessed in chimeric pigs and compared with control pigs. Peripheral blood lymphocytes (PBL) from chimeric pigs demonstrated a wide range of cytolysis (0 to 85%, median 13%) whereas PBL from control pigs were consistently lysed with these conditions (86 to 99%, median 96.5%, P < 0.0001). Accommodation or reduction in cytolysis did not correlate with the amount of chimerism. A longitudinal study demonstrated persistent accommodation of the PBL for as long as 15 weeks, when the donors averaged 68 kg in weight. Accommodation has been induced by low levels of antibodies interacting with the target tissue. An ELISA for sheep IgG was developed and the serum from newborn pigs assessed. Sheep IgG (up to 4.6 microg/ml) was detected in four of seven piglets with chimerism detectable by flow cytometry and in one of four piglets with minimal chimerism, detectable only by PCR. Lymphocyte accommodation was observed in all pigs with detectable sheep IgG. Of four pigs without accommodation, none had sheep IgG. Three pigs without detectable sheep IgG also had accommodation, suggesting that factors other than sheep IgG may induce accommodation. Acute vascular rejection was not observed in the heterotopic heart transplants from six donors with PBL accommodation. Only one incident of moderate diffuse cellular rejection (grade 3) was observed. PMID- 12535228 TI - Xenotransplantation public perceptions: rather cells than organs. AB - The aim of this study was to describe some of the factors that might play a role in influencing attitude to xenotransplantation: first, the consideration of receiving cells and tissue from xenotransplants in relation to whole xeno-organs; secondly, the fact that there is greater uncertainty regarding the result and risk of infection associated with xenotransplantation than with allotransplantation. We also describe the attitude to research on xenotransplantation, and the relationship between the attitude to receiving a xenotransplant and an allotransplant. Finally, we describe the attitude to xenotransplantation in relation to treatment for renal failure and waiting-time for allotransplantation. A questionnaire was sent to randomly selected members of the public aged 18 to 75 (n=1,000) and to all patients in the same age range who were waiting for kidney transplants in Sweden in the spring of 1998 (n=460). The response rate was 60% among the public and 87% among the patients. Both study groups were positive to a greater extent in their attitude to receiving cells and tissue than to receiving a whole organ such as a kidney. The response 'rather positive' to receiving organs was generally favored by the public, whereas the most generally favored response to receiving cells and tissue was 'very positive'. When there was suggested to be a greater uncertainty regarding the outcome with xenotransplantation compared with allotransplantation, the number of negative and uncertain respondents increased, both among the public and the patients. Eighty percent of the public and about 90% of the patients were in favor of continued research on xenotransplantation. Of those members of the public who responded, the attitude to receiving an organ from a human was positive in 86% of cases, with an emphasis on 'very positive'. There was a moderate relation between the attitude to receiving an organ from a human and to receiving a xenotransplant. Among the patients, there was no systematic or strong relation between the attitude to xenotransplantation and the kind of dialysis treatment they were on. Neither was there any systematic or strong relation to the waiting-time. The overall impression is that the attitude to xenotransplantation seems to be most influenced by whether the xenotransplant would involve whole organs or cells and uncertainty regarding the outcome. PMID- 12535229 TI - An exploratory investigation of the effect of arsenic trioxide on anti-Gal antibody production in baboons. AB - BACKGROUND: Arsenic trioxide (As2O3) is an anticancer drug that has been reported to induce apoptosis and inhibit differentiation in human plasmacytoma and normal plasma/B cells without significant myelosuppression. We assessed the ability of As2O3 as single therapy or in combination with an anti-CD20 monoclonal antibody (mAb) and whole body irradiation (WBI) to deplete B and plasma cells, both in vitro and in vivo, and to reduce the level of anti-alphaGal1-3Gal antibody (anti Gal Ab) in baboons. METHODS: In vitro the effect of As2O3 on antibody secretion (anti-Gal IgM, total IgG and IgM) was measured by enzyme-linked immunospot assay (ELISPOT). Its inhibition of proliferation of baboon splenocytes and the NCI-H929 human plasmacytoma cell line was measured by tritiated thymidine uptake. In vivo: all baboons (n=7) had undergone splenectomy. The effects of As2O3 (0.18 to 0.36 mg/kg) on B/plasma cell depletion and anti-Gal Ab production were assessed in three baboons. For comparison, three baboons received either WBI (2 x 150 cGy) or anti-CD20 mAb (20 mg/kg x 4 doses), or both WBI and anti-CD20 mAb. A final baboon received As2O3 + WBI (150 cGy) + anti-CD20 mAb. Anti-Gal Ab levels were measured daily by ELISA. Depletion of B cells from blood and bone marrow (BM) was monitored by flow cytometry and by histology of lymph nodes (LN). Autopsy was performed in three baboons. RESULTS: In vitro: As2O3 (at 5 x 10-6 mol/l) reduced anti-Gal IgM and total IgM secretors by 76% (P=0.53) and 95% (P < 0.001), respectively, but did not reduce total IgG secretors. As2O3 inhibited in a dose dependent manner the proliferation of activated splenocytes and of the NCI-H929 plasmacytoma cell line; complete inhibition was achieved at a dose of 1 x 10-5 mol/l. In vivo: As2O3 was found to be toxic at the doses given and was associated with the deaths of two of the four baboons that received it. Daily intravenous therapy with As2O3 alone reduced B cells (CD20+) in the blood (by 50 to 90%), BM (40%) and LN (20 to 30%), but anti-Gal Ab levels were not significantly decreased. Anti-CD20 mAb therapy alone or WBI alone depleted B cells by 100% in the blood and BM, and 80 to 100% in the LN. The combination of anti-CD20 mAb + WBI led to depletion of B cells in blood, BM and LN for 3 months, but reduction of anti-Gal Ab remained marginal. The combination of As2O3 + anti-CD20 mAb + WBI did not reduce anti-Gal Ab levels further. At autopsy in the latter baboon, B cells remained present in Peyer's patches and tonsils. CONCLUSIONS: In vitro: As2O3 reduced B/plasma cell numbers and suppressed IgM secretors, but not IgG secretors. In vivo: As2O3 was not as effective as either anti-CD20 mAb or WBI in depleting B/plasma cells, and was largely ineffective in reducing anti-Gal Ab levels. Its administration was associated with considerable toxicity. Autopsy in one baboon suggested that B cells in Peyer's patches and tonsils may be resistant to therapy and remain a source of continuing production of anti-Gal Ab. PMID- 12535231 TI - Value in health: "bridging the gap" among our disciplines. PMID- 12535230 TI - An investigation of the specificity of induced anti-pig antibodies in baboons. AB - AIM: To provide information on the specificity of induced anti-pig antibodies (Abs) in baboons after exposure and sensitization to pig antigens. MATERIALS AND METHODS: Baboons (n=7) received either porcine mobilized peripheral blood progenitor cells (n=3), kidney (n=3) or heart (n=1) transplants. After rejection of these cells or organs, pre- and post-rejection sera were analyzed by enzyme linked immunosorbent assay (ELISA) and flow cytometry to detect and measure anti Galactosealpha1,3Galactose (Gal) and anti-non-Gal Abs. To study the anti-non-Gal carbohydrate response, the sera were incubated with pig red blood cells pretreated with alpha-galactosidase (to remove Gal) and three other exoglycosidases to remove other potential oligosaccharide epitopes, and studied by flow cytometry. To study the anti-swine leukocyte antigen (SLA) response, non Gal Abs from two baboons sensitized with kidneys from inbred miniature swine of dd or aa haplotype, respectively, were adsorbed on cells of aa, cc, or dd haplotypes, and binding to aa, cc or dd cells was measured by flow cytometry. Cytotoxicity of anti-non-Gal Abs was tested in vitro by a complement-mediated cytotoxicity assay, using pig cells as targets. RESULTS: In pre-transplant and pre-rejection sera, anti-Gal Abs were detected, but anti-non-Gal Abs were either absent or at minimal levels. After exposure to pig antigens, baboons developed induced anti-Gal and anti-non-Gal Abs. No anti-non-Gal Abs directed to the tested carbohydrate epitopes could be detected. Anti-non-Gal Abs showed minor evidence of specific SLA haplotype reactivity, suggesting that the major Ab response was to pan-pig determinants. Anti-non-Gal Abs showed a low level of complement mediated lysis of pig cells in vitro. CONCLUSIONS: In this limited study, no Ab response to non-Gal carbohydrates was observed, and anti-SLA specificity was minor, indicating that most induced anti-non-Gal Ab was directed against non specific pig proteins, including SLA-epitopes. PMID- 12535232 TI - Making decisions on technology availability in the British National Health Service--why we need reliable models. PMID- 12535233 TI - The ISPOR Good Practice Modeling Principles--a sensible approach: be transparent, be reasonable. PMID- 12535234 TI - Principles of good practice for decision analytic modeling in health-care evaluation: report of the ISPOR Task Force on Good Research Practices--Modeling Studies. AB - OBJECTIVES: Mathematical modeling is used widely in economic evaluations of pharmaceuticals and other health-care technologies. Users of models in government and the private sector need to be able to evaluate the quality of models according to scientific criteria of good practice. This report describes the consensus of a task force convened to provide modelers with guidelines for conducting and reporting modeling studies. METHODS: The task force was appointed with the advice and consent of the Board of Directors of ISPOR. Members were experienced developers or users of models, worked in academia and industry, and came from several countries in North America and Europe. The task force met on three occasions, conducted frequent correspondence and exchanges of drafts by electronic mail, and solicited comments on three drafts from a core group of external reviewers and more broadly from the membership of ISPOR. RESULTS: Criteria for assessing the quality of models fell into three areas: model structure, data used as inputs to models, and model validation. Several major themes cut across these areas. Models and their results should be represented as aids to decision making, not as statements of scientific fact; therefore, it is inappropriate to demand that models be validated prospectively before use. However, model assumptions regarding causal structure and parameter estimates should be continually assessed against data, and models should be revised accordingly. Structural assumptions and parameter estimates should be reported clearly and explicitly, and opportunities for users to appreciate the conditional relationship between inputs and outputs should be provided through sensitivity analyses. CONCLUSIONS: Model-based evaluations are a valuable resource for health care decision makers. It is the responsibility of model developers to conduct modeling studies according to the best practicable standards of quality and to communicate results with adequate disclosure of assumptions and with the caveat that conclusions are conditional upon the assumptions and data on which the model is built. PMID- 12535235 TI - How well have practices followed guidelines in prescribing antihypertensive drugs: the role of health insurance. AB - BACKGROUND: The US Joint National Committee (JNC) on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure issues guidelines on the optimal first-line drug therapy in treating hypertension. Despite broad dissemination of these guidelines, prescribing practices have long remained discrepant with recommendations. The purpose of this study was to examine the role of insurance type in the selection of drugs for hypertension treatment in light of the JNC guidelines. METHODS: Subjects were derived from the 1996 Medical Expenditures Panel Survey who had a diagnosis of essential hypertension and who were prescribed a diuretic, beta-blocker, calcium channel blocker (CCB), or ACE inhibitor (ACEI) as monotherapy. Using the nationally representative sample, this study presents the first estimates of the impact of insurance policies on the choice of antihypertensive drugs while controlling for predisposing, enabling, and need variables in the context of a logistic health-care utilization model. RESULTS: Nationally in 1996, more than twice as many subjects (7.3 million) were taking ACEIs or CCBs compared to diuretics or beta-blockers (3.1 million) as the first-line drug therapy, a sharp contrast to the JNC guidelines. Patients with health maintenance organization (HMO) insurance were much less likely than fee for service (FFS) patients to follow the JNC guidelines in this respect (odds ratio 0.50, P <.01), controlling for all other factors. Individuals with all other public insurance and no insurance were not statistically different from the FFS group in the use of the study drugs. Other significant factors in the regression model were being of African American descent, being unmarried, having higher out-of-pocket payment, being in excellent physical health, having diabetes, and being diagnosed with essential hypertension after 1988. Each was associated with a decreased likelihood of following the JNC recommendations for the use of diuretics or beta-blockers. CONCLUSIONS: After controlling for other predisposing, enabling, and need variables, patients who had HMO coverage were significantly more likely than FFS patients to receive ACEIs or CCBs. Given a popular public perception of HMOs being most cost conscious in providing health care, further research is needed to understand why prescribing patterns associated with HMOs have poorly followed the JNC recommendations. PMID- 12535236 TI - Computerized health information and the demand for medical care. AB - OBJECTIVE: Consumer health information, once the domain of books and booklets, has become increasingly digitized and available on the Internet. This study assessed the effect of using computerized health information on consumers' demand for medical care. METHODS: The dependent variable was self-reported number of visits to the doctor in the past year. The key independent variable was the use of computerized health information, which was treated as endogenous. We tested the effect of using computerized health information on physician visits using ordinary least squares, instrumental variables, fixed effects, and fixed-effects instrumental variables models. The instrumental variables included exposure to the Healthwise Communities Project, a community-wide health information intervention; computer ownership; and Internet access. Random households in three cities were mailed questionnaires before and after the Healthwise Communities Project. In total, 5909 surveys were collected for a response rate of 54%. RESULTS: In both the bivariate and the multivariate analyses, the use of computerized health information was not associated with self-reported entry into care or number of visits. The instrumental variables models also found no differences, with the exception that the probability of entering care was significantly greater with the two-stage conditional logit model (P <.05). CONCLUSIONS: Although providing people with health information is intuitively appealing, we found little evidence of an association between using a computer for health information and self-reported medical visits in the past year. This study used overall self-reported utilizations as the dependent variable, and more research is needed to determine whether health information affects the health production function in other important ways, such as the location of care, the timing of getting care, or the intensity of treatment. PMID- 12535237 TI - Functional status and somatization as predictors of medical offset in anxious and depressed patients. AB - OBJECTIVE: Certain anxious/depressed primary care patients decrease medical utilization after mental health treatment. Previous research has established demo graphic and medical comorbidities as distinguishing these patients. We asked whether characteristics such as symptom severity, somatization, or health-related quality of life (HRQoL) could also distinguish patients who reduce or increase primary care utilization after mental health care. METHODS: Primary care patients in a mixed-model HMO were screened for untreated anxiety with and without depression, using the Symptom Checklist (SCL-90-R) and medical records abstractions, and also for HRQoL (SF-36). We identified 165 symptomatic patients who subsequently received mental health treatment and then defined two subgroups: 1) offset patients (reduced medical utilization the year after initiation of mental health treatment) (N=97); and 2) no-offset patients (increased utilization) (N =68). RESULTS: Three HRQoL domains (general health perceptions, physical functioning, and role functioning- physical) predicted increased offset savings in the year after initiation of mental health treatment. Each point of improved functioning in these domains was associated with 4 dollars to 10 dollars of additional offset savings. Somatization-related comorbidities were predictive of greater additional costs (230 dollars). CONCLUSION: Using models to predict individual patient costs, we found that HRQoL and somatic comorbidities did not predict by anxiety/depression symptom severity or medical comorbidities, but by increasing or decreasing utilization after mental health care. Patients with higher functioning levels and no somatic comorbidities were most likely to reduce utilization. These findings support growing evidence for the need of inclusion of reliable indicators of somatization and patients' functioning in offset research and inpatient care. PMID- 12535238 TI - Drug-related problems and quality of life in arthritis and low back pain sufferers. AB - OBJECTIVE: The objective of this study was to determine the relationship between drug-related problems (DRPs) and health-related quality-of-life (HRQoL) in ambulatory, community-dwelling patients with musculoskeletal disorders. METHODS: A 12-month, prospective, observational study was conducted in 12 independent community pharmacies in eastern Iowa. Ambulatory patients with self-reported diagnoses of osteoarthritis, rheumatoid arthritis, or low back pain were invited to participate. During quarterly visits to the pharmacy, patients used touch screen computers to fill out the Short Form-36 (SF-36) general health survey. Using the results of these point-of-service health status assessments, community pharmacists interviewed patients to assess for DRPs. To examine the influences of different DRP characteristics on HRQoL and controlling for potential confounders, both univariate and multivariate analyses were performed using the change in physical component summary (PCS) score and mental component summary (MCS) score of the SF-36 from baseline to 12 months as the dependent variables. In each regression, the independent variables were those significant variables from the univariate analyses, as well as the types of DRPs and their outcomes. RESULTS: A total of 461 patients were enrolled in the study. Through 12 months, 926 cumulative DRPs were identified. Overall regression models were significant for the PCS and MCS scores, respectively. Two types of DRPs showed significant negative associations with change in PCS: wrong drug and needs additional drug therapy. One type of DRP showed significant negative association with change in MCS: needs additional drug therapy. Resolution or improvement in DRPs showed a significant positive correlation with change in MCS but not PCS. CONCLUSIONS: Two DRPs, needs additional drug therapy and wrong drug, are associated with reduced self-reported physical health in arthritis and low back pain, while the DRP needs additional drug therapy is also associated with reduced self-reported mental health. Resolution of DRPs is associated with improvement in mental health in this cohort. PMID- 12535239 TI - Economic burden of long-term complications of deep vein thrombosis after total hip replacement surgery in the United States. AB - BACKGROUND: Estimates of the cost of long-term complications of a primary deep vein thrombosis (DVT), including the post-thrombotic syndrome (PTS) and recurrent venous thromboembolism (VTE), may be relevant for resource allocation decisions. OBJECTIVE: The objective of this study was to provide US cost estimates of the long-term complications of a primary DVT, which occurs in approximately 5% to 20% (with adequate thromboprophylaxis) and 50% (in the absence of thromboprophylaxis) of total hip replacement surgeries (THRS). METHODS: A literature-based model was used to project the excess long-term complication costs of DVT following THRS. The model simulated the natural history of DVT complications using published estimates of the incidence and prognosis of PTS and recurrent VTE. Each complication was assigned a cost obtained by multiplying the amount of resources used in its management by the unit price of these resources. RESULTS: The annual per-patient cost of each complication was as follows: mild-to-moderate PTS, 839 dollars in the first year and 341 dollars in subsequent years; severe PTS, 3817 dollars in the first year and 1677 dollars in subsequent years; DVT, 3798 dollars; and pulmonary embolism, 6604 dollars. The average discounted lifetime cost of DVT complications was estimated to be 3069 dollars (95% interval 2091 dollars-4279 dollars). CONCLUSIONS: The long-term complications of a primary DVT represent a significant economic burden. Preventing a DVT could arguably lead to substantial savings in long-term DVT complications. PMID- 12535240 TI - Costs of asthma in a cohort of Swiss adults: associations with exacerbation status and severity. AB - OBJECTIVE: This article will address the effect of exacerbation status, disease severity (defined by medication required), and other variables on resource use and costs for asthma in Switzerland in 1996 to 1997. METHODS: A retrospective chart-based study was performed. A sample of 422 adults was analyzed. Target variables were stratified by disease severity and exacerbation status. Bivariate associations were assessed. Multiple linear regression was performed on the logarithm of direct medical costs. RESULTS: The probability of exacerbations was positively associated with disease severity. Resource use and costs were associated with both of these variables. Multiple linear regression identified age, presence of asthma-related comorbidities, degree of severity, exacerbation status, quick reliever versus controller therapy, and diagnosis or treatment by a pulmonologist as independent influences on direct costs. An interaction between severity and exacerbation status was also noted. Regression identified direct costs in the highest severity group to be 2.5 times higher than those in the lowest group, if there were no exacerbations. If exacerbations were present, costs were 5.7 times higher. CONCLUSIONS: Because of its high prevalence, asthma has a high impact on public health. This impact depends on disease severity and, according to these findings, may also depend on the extent to which exacerbations are avoided or at least controlled. PMID- 12535241 TI - Vaccination politics. PMID- 12535242 TI - Plasmodium falciparum multiple infections in Mozambique, its relation to other malariological indices and to prospective risk of malaria morbidity. AB - We describe the frequency of Plasmodium falciparum clones infecting individuals living in a rural area of southern Mozambique and analyse the relationship between multiplicity of infection, age and other malariometric indices, including prospective risk of clinical malaria. The genotyping was based on the use of restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) analysis of P. falciparum merozoite surface protein 2 (msp2). We analysed 826 samples collected during five cross-sectional surveys from residents of Manhica ranging in age from 4 months to 83 years. We also determined the multiplicity of infection in samples obtained from 6-month-old infants (n = 79) and children <10 years (n = 158) who were then treated and followed prospectively for 1 year or 75 weeks, respectively. Multiplicity of infection did not vary significantly during the first year of life, but increased thereafter, and decreased during adulthood to the levels found in infants. With increasing multiplicity of infection, there was a statistically significant decrease in the risk of submicroscopic infections. There was also a significant correlation between multiplicity of infection and parasite density in infants, children <4 years of age and adults, suggesting that high densities increase the probability of discriminating more clones in complex infections. We found that the relationship between multiple infections and malaria morbidity is age-dependent. In infants, the risk of subsequent episodes of clinical malaria was related to the parasite density but not to baseline multiplicity of infection. In older children, however, the more clones a child carried, the more likely they were to have a clinical malaria episode, and this was true after adjusting for parasite densities. This change in the association between multiplicity and risk of clinical malaria may indicate a shift in the host response to P. falciparum. PMID- 12535243 TI - Seizure disorders among relatives of Kenyan children with severe falciparum malaria. AB - PURPOSE: The cause of seizures in children with falciparum malaria is unclear. In malaria endemic areas, children who develop severe falciparum malaria with seizures may have a genetically higher risk of epilepsy or febrile seizures. We used the history of seizures in relatives of children previously admitted with malaria to determine if there is evidence for a familial predisposition of seizures in children admitted with malaria and seizures or cerebral malaria. METHODS: Family history of seizures were obtained from the parents/guardians of 81 children (35 children previously admitted with severe malaria and 46 children matched for age who had not been admitted with severe malaria). Data were collected on frequency, duration, age of onset, presence of fever and causes of seizures. RESULTS: The prevalence of seizures in the relatives of children not admitted with severe malaria was 4.3%, of whom 2.2% had a history of seizures compatible with febrile seizures, and 1.1% with epilepsy. Overall the odds ratio (OR) for relations of children admitted with malaria, to have a seizure disorder was 1.41 [95% confidence interval (CI) 1.06-1.88]. There was a significant risk of the relatives dying if they had epilepsy [relative risk 1.88 (95% CI 1.11 3.19)], but not for other seizure disorders (i.e. febrile, single or unclassifiable seizures). CONCLUSION: Relatives of children admitted with severe falciparum malaria are more likely to have a seizure disorder compared with controls, but it is unclear if this is because of a genetic propensity or caused by exogenous factors such as malaria. PMID- 12535244 TI - Malaria and amphetamine 'horse tablets' in Thailand. AB - During recent clinical malaria research in Thailand we found a high frequency of amphetamine misuse and withdrawal amongst patients admitted to hospital with Plasmodium falciparum malaria. This comorbidity may cause diagnostic confusion, alter malaria pathophysiology and lead to drug interactions. PMID- 12535245 TI - Therapeutic efficacy of chloroquine plus sulphadoxine/ pyrimethamine compared with monotherapy with either chloroquine or sulphadoxine/pyrimethamine in uncomplicated Plasmodium falciparum malaria in Laos. AB - In a southern border province of Lao PDR, we compared the efficacy of antimalarial drug combinations in patients aged >or=1 year with uncomplicated Plasmodium falciparum malaria: monotherapy with either mefloquine (MQ), chloroquine (CQ), or sulphadoxine/pyrimethamine (SP) vs. the combination of both CQ and SP. Follow-up time was 14 days. Of 265 P. falciparum positive patients, 119 were enrolled in the drug trial. Significantly more patients treated with CQ than with SP developed early or late treatment failure [44.8%vs. 17.9%, relative risk (RR) = 2.51, 95% CI 1.03-6.12]. In the SP group, 82.1% were sensitive and 17.9% were treatment failures. The combination treatment CQ plus SP resulted in 83.3% sensitivity and 16.7% treatment failures. Combination treatment has no advantage over monotherapy with SP (RR = 1.01, 95% CI 0.8-1.3). All patients who received MQ for treatment (total dose 25 mg/kg) were cured within the 14 days of follow-up. The findings of this study suggest that use of CQ as first-line treatment of uncomplicated malaria in the Lao PDR has to be reconsidered. The combination of both CQ and SP has been discussed as a cost-effective alternative treatment, but in our patient population achieved no better results than single therapy with SP. PMID- 12535246 TI - In vitro activity of chloroquine, quinine, mefloquine and halofantrine against Gabonese isolates of Plasmodium falciparum. AB - OBJECTIVES: To determine the in vitro activity of antimalarial drugs against isolates of Plasmodium falciparum in Gabon. METHODS: Plasmodium falciparum isolates were collected from symptomatic infections in the hospitals of Bakoumba and Franceville, south-east Gabon and in 2000. In vitro activity of chloroquine, quinine, mefloquine, halofantrine was measured by the isotopic microtest. RESULTS: A total of 60 and 62 isolates gave interpretable data in Franceville and Bakoumba, respectively. In Franceville, 50.0% (mean IC50 = 111.7 nm), 0% (mean IC50 = 156.7 nm), and 21.2% (mean IC50 = 12.4 nm) of isolates, respectively, showed in vitro resistance to chloroquine, quinine and mefloquine. In Bakoumba, we saw resistance to chloroquine, quinine, mefloquine and halofantrine in 95.0% (mean IC50 = 325.8 nm), 10.2% (mean IC50 = 385.5 nm), 47.5% (mean IC50 = 24.5 nm) and 18.2% (mean IC50 = 1.9 nm) of isolates, respectively. Activities of chloroquine and mefloquine, chloroquine and quinine, and mefloquine and quinine were positively correlated. CONCLUSIONS: Antimalarial drug resistance is high in this area of Gabon. The extent of resistance is disparate, as all tested drugs were less efficacious in Bakoumba than in Franceville. PMID- 12535247 TI - Myocardial damage in falciparum malaria detectable by cardiac troponin T is rare. AB - OBJECTIVES: To estimate the prevalence of myocardial damage in falciparum malaria by serum concentration of cardiac troponin T. METHODS: Retrospective study of stored sera and patient files; assessment of acute myocardial damage by serum concentration or activity of cardiac troponin T, creatine kinase, creatine kinase MB and myoglobin and by routine electrocardiography. RESULTS: A total of 161 patients with falciparum malaria were included in the study; troponin T was elevated in one case (0.6%), no CK-MB elevations were found, myoglobin was elevated in 10 of 161 patients (6.2%), all of whom were elderly and had concomitant elevated serum concentration levels of cystatin C; ECG abnormalities were seen in 23 patients. CONCLUSION: Assessed by troponin T, myocardial damage in falciparum malaria is rare. PMID- 12535248 TI - Lower levels of the circulating neuropeptide somatostatin in Schistosoma mansoni infected patients may have pathological significance. AB - In recent years, cases of severe morbidity (fibrosis, haematemesis, hepatosplenomegaly, ascites) caused to Schistosoma mansoni infections are on the rise in Northern Senegal. The neuropeptide somatostatin is reported to decrease portal pressure, control variceal bleeding and fibrosis, and reduce Schistosoma caused clinical morbidity in the rodent model. The aim of this study was to delineate the role of somatostatin in S. mansoni-caused pathogenesis, by studying host levels of somatostatin in the peripheral blood of uninfected and S. mansoni infected individuals. Subjects from the district dispensary at Richard Toll, in the Medical Region of Saint-Louis, Senegal, infected with S. mansoni and suffering from severe morbidity were selected. A separate group consisted of individuals resident in the same region but uninfected at the time of the study. Significantly lower somatostatin levels were detected in severe morbidity patients, compared with the exposed but uninfected subjects. In patients with schistosomiasis physiological levels of somatostatin may determine disposition of particular individuals towards severe morbidity, as opposed to others. Host pathology can thus be alleviated by the therapeutic ability of somatostatin to treat bleeding oesophageal varices, reduce portal pressure and prevent progression to severe fibrosis. PMID- 12535249 TI - Efficacy and efficiency of new Bacillus thuringiensis var israelensis and Bacillus sphaericus formulations against Afrotropical anophelines in Western Kenya. AB - We evaluated the efficacy of new water-dispersible granular (WDG) formulations of Bacillus thuringienis var. israelensis (Bti; VectoBac) and B. sphaericus (Bs; VectoLex), Valent BioScience Corp., Illinois, USA) for the control of larval Anopheles gambiae sensu lato Giles mosquitoes in a malaria-endemic area around Lake Victoria, Western Kenya. WDG and powder formulations were compared in laboratory bioassays and followed by efficiency and residual effect assessments of both WDG formulations in open field experiments. LC50 and LC95 values for the Bti/Bs strains and their formulations show high susceptibility of A. gambiae sensu stricto under laboratory conditions. The larvae proved more susceptible to Bs than to Bti and the WDG formulations were slightly superior to the powder formulations. High efficiency was also shown in the open field trials, and a minimum dosage of 200 g/ha Bti WDG, representing the LC95 of the laboratory tests, was sufficient to fully suppress emergence of mosquitoes when applied at weekly intervals. Bti WDG did not show a residual effect, irrespective of the concentration applied. The Bs WDG formulation, however, showed significant larval reductions up to 11 days post-treatment at application doses of either 1 or 5 kg/ha. We conclude that the main malaria vector in our study area is highly susceptible to these microbial control agents. Minimum effective dosages to achieve elimination of the larval population in a given habitat are extremely low and environmental impact is negligible. Microbial products for larval control have therefore great potential within Integrated Vector Management programmes and may augment control efforts against adult vector stages, such as the use of insecticide-treated bednets, in many parts of Africa. PMID- 12535250 TI - Malaria control in complex emergencies: the example of East Timor. AB - Malaria control in complex emergencies forms part of the World Health Organization's Roll Back Malaria (RBM) initiative. The underlying principle is a partnership with other UN agencies and non-governmental organizations, with the RBM programme providing support to its implementing partners through a Technical Support Network. This innovative approach was applied for the first time in 1999 2000, following the return of stability and security to East Timor. The RBM programme assessed the malaria situation during the acute emergency phase and identified programme priorities. Two non-governmental organizations were subsequently invited to operate as implementing partners. Individual proposals were developed and funding obtained, but no overall organizational and planning framework was established. Implementation commenced quickly, addressing aspects of prevention, diagnosis and treatment. As East Timor progressed into the transitional phase towards independence, the programme was not realigned to the changing context. Absence of monitoring and evaluation was a significant factor contributing to the resulting continuation of emergency malaria control activities well into the transitional phase. East Timor's example of malaria control in complex emergencies provides important lessons: (i). partnership roles and responsibilities should be clearly defined from the beginning of the programme, (ii). planning, monitoring and evaluation should be instigated from the start with the aim to develop long-term strategies and policies, (iii). expert support is necessary to guide implementation at all stages, (iv). the flexibility to react to changing priorities should be ensured as the context of the emergency and the available health structure changes, and (v). the implementation process, and alternatives for continuation of activities once the RBM complex emergency partnership has terminated, should be clarified to all partners involved. PMID- 12535251 TI - Treatment seeking behaviour and costs due to acute and chronic forms of lymphatic filariasis in urban areas in south India. AB - Previous estimates on the economic burden of lymphatic filariasis (LF) in India and elsewhere were primarily based on studies in rural areas. We investigated the treatment costs due to acute and chronic forms of LF in urban areas, where nearly one-third of the affected people live. Almost 98% of the patients with acute episodes of adenolymphangitis (ADL) underwent treatment and 49% of chronic patients also received treatment. The average treatment cost per ADL episode (n = 108) was Rs 22.21 +/- 53.84 (US dollars 0.46 +/- 1.12). The overall (n = 200) treatment costs incurred by a chronic patient per visit were Rs 16.71 +/- 62.36 (US dollars 0.35 +/- 1.30); for those who paid (n = 98) they were Rs 34.10 +/- 85.90 (US dollars$ 0.71 +/- 1.79). These costs are considerably higher than in rural areas. Government health centres and private practitioners were important sources of treatment. Treatments received from private practitioners were considerably more expensive than those from government health facilities. The cost of medicine accounted for 44% and 50% of the total expenditure on treatment for acute and chronic disease patients, respectively. The medical personnel from these treatment sources need to be trained on the new morbidity management methods, which are likely to be more effective than the current methods of treatment. PMID- 12535252 TI - The absence of HIV seropositivity contrasts with a high prevalence of markers of sexually transmitted infections among registered female sex workers in Toliary, Madagascar. AB - In a cross-sectional study in 1998 we assessed human immunodeficiency virus (HIV) and syphilis infections and their risk factors among the 316 registered female sex workers (FSWs) of Toliary, south-west Madagascar. No case of HIV infection was detected, but 18.4% of registered FSWs had syphilis. Only half of these women regularly used condoms. In a multiple logistic regression analysis, risk factors for syphilis infection were multiple clients per week and, paradoxically, regular use of condoms. The variables associated with irregular use of condoms were younger ages of registered FSWs, multiple clients per week and Malagasy clients. The high prevalence of syphilis infection associated with irregular use of condoms might facilitate a very fast spread of HIV infection among these FSWs. Promotion of condom use and surveillance of sexually transmitted infections and HIV infection incidence are needed in the south of Madagascar. PMID- 12535253 TI - Health and nutritional status of orphans <6 years old cared for by relatives in western Kenya. AB - One of the consequences of the HIV/AIDS epidemic in sub-Saharan Africa is the increase in the number of orphans, estimated to have reached 6-11% of children <15 years old in 2000. Orphans who stay in their communities may be at increased risk for poor health due to reduced circumstances and loss of parental care. We have used data from a population-based study in rural western Kenya to compare basic health and nutritional indicators between non-orphaned children <6 years old and children who lost either or both of their parents. In June 2000, all children <6 years old who had been recruited for a cross-sectional survey in 60 villages of Rarieda Division, western Kenya, in June 1999 were invited to return for a follow-up survey. Basic demographic characteristics, including the vital status of the child's parents, and health histories were requested from all 1190 participants of the follow-up survey, along with a finger-prick blood sample for determination of malaria parasite status and haemoglobin (Hb) levels. Height-for age (H/A) and weight-for-height (W/H) Z-scores were also calculated from anthropometric measurements. Overall, 7.9% of the children had lost one or both their parents (6.4% had lost their father, 0.8% had lost their mother and 0.7% had lost both parents). While there was no difference between orphans and non orphans regarding most of the key health indicators (prevalence of fever and malaria parasitaemia, history of illness, Hb levels, H/A Z scores), W/H Z-scores in orphans were almost 0.3 standard deviations lower than those of non-orphans. This association was more pronounced among paternal orphans and those who had lost a parent more than 1 year ago. These results suggest that the health status of surviving orphans living in their community is similar to that of the non orphan population, but longitudinal cohort studies should be conducted to determine better the overall impact of orphanhood on child health. PMID- 12535254 TI - Hybridoma cell agglutination as a novel test to detect circulating antigen of Schistosoma japonicum. AB - We developed a serodiagnostic test which is based on the agglutination of hybridoma cells. In the presence of specific antigen, agglutination of the fixed and stained cells occurs and can be visualized in analogy to traditional erythrocyte agglutination. The procedures were developed with a murine cell line producing a monoclonal antibody against a schistosome gut protein and sera of patients and mice infected with Schistosoma japonicum. This test is capable of detecting circulating antigen during pre-patency in mice infected with 50 cercariae. Its sensitivity was high with acute schistosomiasis japonica (97%, n = 32) and moderate with chronic cases (75%, n = 57). No positive reactions were obtained with healthy persons (n = 78) or patients infected with other parasites (Chlonorchis sinensis, n = 20; Paragonimus westermani, n = 20; Plasmodium vivax, n = 10) or suffering from lupus erythomatodus (n = 5) or mononucleosis (n = 10). PMID- 12535255 TI - Urinary schistosomiasis in preschool children in settlements around Oyan Reservoir in Ogun State, Nigeria: implications for control. AB - We determined the prevalence of urinary schistosomiasis in preschool children below the age of 5 years in three settlements around Oyan Reservoir in Ogun State, Nigeria. Of 209 children screened, 150 (71.8%) had an infection, with no significant difference between males and females; 42.9% of infants were infected. Both prevalence and intensity of infection increased significantly with age (P < 0.005). Most (62.7%) infections were light (<50 eggs/10 ml urine). A 17.7 percentage of the children had visible haematuria, which increased with age (P < 0.005). Focus group discussions (FGDs) with adult men and women revealed that infection in preschool children was primarily because of exposure occasioned by the mothers' domestic (washing and bathing) and occupational (fishing) activities, while older children could go swimming on their own. Although the participants claimed that using a different water supply may not be effective in combating the disease, as their entire existence was tied to the reservoir, we propose that health education geared towards changing behaviour and attitudes is necessary. As preschool children are a source of both contamination and transmission, control programmes must take them into account. PMID- 12535256 TI - Profile of eye lesions and vision loss: a cross-sectional study in Lusambo, a forest-savanna area hyperendemic for onchocerciasis in the Democratic Republic of Congo. AB - OBJECTIVE: To determine baseline data regarding eye lesions and vision loss in five villages of Lusambo, an onchocerciasis-hyperendemic forest-savanna area in the Democratic Republic of Congo (DRC), in preparation of mass ivermectin distribution. METHODS: Five villages were selected by simple randomization. Through a cross-sectional design, 750 subjects were examined ophthalmologically. The eye examination included acuity visual measurement, slit-lamp examination, ophthalmoscopy, intraocular pressure measurement, and visual field assessment by the Wu-Jones test. RESULTS: There was a high prevalence of onchocerciasis-related eye lesions compared with non-onchocercal lesions. Chorioretinitis (20%) was the most frequent disease, others were punctate keratitis and microfilariae in the anterior chamber in equal frequency (13.8%), white intraretinal deposits (10.4%) and iridocyclitis (8%). Vision loss was discovered in 8.5% of the subjects, of whom 0.5% had bilateral blindness, 2.2% had monocular blindness and 5.7% had visual impairment. Vision loss was mostly caused by onchocerciasis-related diseases, especially those affecting the anterior segment of the eye. CONCLUSION: Features of ocular onchocerciasis usually described in forest and savanna areas were both found in this forest-savanna zone of the DRC. PMID- 12535257 TI - Assessment of functional limitations caused by Mycobacterium ulcerans infection: towards a Buruli ulcer functional limitation score. AB - The purpose of this study of treated Buruli ulcer patients in Ghana was to identify and assess late sequelae of treated Buruli ulcer using a goniometer, and to develop a scoring system for functional limitations. Of 78 patients, 58% (n = 45) had a reduction in the range of motion of one or more joints: 30% (n = 23) had one or more functional limitations of the leg and 21% (n = 16) of the arm; 49% (n = 38) had a functional limitation. Of all patients with affected knees, the predicted average extent of limitation was 63%. In patients with affected ankles, limitation was 78% on average; in those with elbow involvement, this was 76% on average, and in wrists involved, 65%. All of the hands involved were markedly restricted. We propose a simplified and functional scoring system that should be tested for validation in a second patient sample, and, if properly validated and adjusted, can be used in future intervention trials. PMID- 12535258 TI - Influenza and the transplant recipient. PMID- 12535259 TI - The pathogenesis and management of influenza virus infection in organ transplant recipients. AB - Infection with influenza viruses poses specific problems in adult and pediatric organ transplant recipients, including a higher rate of pulmonary and extra pulmonary complications. Also, data suggest that influenza is associated with acute cellular rejection and chronic allograft dysfunction. The main strategy of influenza prevention has been influenza immunization in order to stimulate local and systemic antibodies. However, studies have shown that antibody response to inactivated influenza vaccine is decreased in all groups of organ transplant recipients. A live attenuated influenza virus vaccine is nearing approval in the United States. However, studies are needed in organ transplant recipients to determine whether the live attenuated influenza virus vaccine can enable these patients to mount a protective immune response and what degree of protection or amelioration of illness is provided by such vaccine. It is also important to verify the safety of this vaccine in organ transplant recipients because live virus may cause severe disease in these patients. Therefore, other modalities of prevention against influenza, such as chemoprophylaxis with antiviral drugs, should be considered in this patient population. The current review provides an overview of the incidence, clinical manifestations, and strategies for the prevention and management of influenza in organ transplant recipients. PMID- 12535260 TI - Cryptococcal meningitis: an analysis among 5,521 consecutive organ transplant recipients. AB - Cryptococcal meningitis has been reported to be an important cause of morbidity and mortality in renal transplant recipients. However, additional studies of recipients of other organ transplants suggested that these patients might be at low risk for cryptococcal meningitis. We examined the incidence and clinical features of cryptococcal meningitis among different groups of organ transplant patients at the University of Pittsburgh Medical Center. From January 1989 through July 1999, 28 patients were diagnosed with cryptococcal meningitis among 5,521 transplant recipients. These included liver (11/2,539), heart (8/372), kidney (7/2,122), lung (1/432), and small bowel (1/56) recipients. The incidence of cryptococcal meningitis was higher in heart and small bowel recipients compared to other transplant populations (P = 0.005). The cryptococcal meningitis related mortality in transplant recipients was 50% and was associated with altered mental status (P = 0.001), absence of headache (P = 0.02), and liver failure (P = 0.002). Multivariable analysis indicated that liver failure was the only independent risk factor for poor prognosis (P = 0.043). All cases of liver failure occurred among liver transplant recipients. Cryptococcal meningitis is associated with significant mortality among organ transplant recipients. The presence of allograft failure in liver transplant recipients with cryptococcal meningitis may be an indicator of poor prognosis in this patient population. PMID- 12535261 TI - Clinical characteristics of 13 solid organ transplant recipients with ganciclovir resistant cytomegalovirus infection. AB - BACKGROUND: Ganciclovir-resistant (GCV-R) cytomegalovirus (CMV) is now being reported with increasing frequency in solid organ transplant recipients. OBJECTIVE: To describe the clinical characteristics and outcomes of all solid organ transplant patients with GCV-R CMV seen between 1990 and 2000 at a single center. METHODS: Patients with clinically suspected GCV resistance had viral isolates subjected to phenotypic analysis by plaque reduction assay, and also genotypic analysis. Medical records of the 13 patients with GCV-R CMV were reviewed for demographic, microbiologic, clinical, and pathologic data. RESULTS: Thirteen patients were identified, including 5 kidney, 1 heart, and 7 lung transplant recipients. All but one patient (92%) were CMV donor seropositive, recipient negative (D+/R-), and 11/13 (85%) had tissue-invasive CMV. CMV viremia was recurrent in 9/13 (69%); in 2 others, the first CMV episode was fatal. Overall, 9/13 (69%) of patients have died, all of CMV or its complications. Of the 10 who received foscarnet, only one survived. All patients had received GCV based prophylactic regimens; 8/13 patients (62%) had received CMV hyperimmune globulin (CMVIG) as part of prophylaxis, 6/13 (46%) had received oral ganciclovir, and 5/13 (38%) had received intermittent (3 x/week) IV ganciclovir for prophylaxis. CONCLUSIONS: GCV-R CMV is associated with CMV D+/R- status, tissue-invasive disease, and high mortality even with foscarnet therapy. Exposure to less than fully therapeutic levels of GCV, in the form of oral or intermittent IV GCV, is common. The use of CMVIG in prophylaxis does not appear to prevent resistance. Further work remains to be done to elucidate the risk factors and optimal mode of prophylaxis and treatment for GCV-R CMV. PMID- 12535262 TI - The incidence of invasive aspergillosis among solid organ transplant recipients and implications for prophylaxis in lung transplants. AB - BACKGROUND: Invasive aspergillosis (IA) is associated with significant morbidity and mortality in solid organ transplant recipients but data on the incidence rates stratified by type of solid organ are limited. OBJECTIVE: To describe the attack rates and incidence of IA in solid organ transplant recipients, and the impact of universal Aspergillus prophylaxis (aerosolized amphotericin B or oral itraconazole) in lung transplant recipients. PATIENTS: The 2,046 patients who received solid organ transplants at the Cleveland Clinic Foundation from January 1990 through 1999 were studied. METHODS: Cases were ascertained through computerized records of microbiology, cytology, and pathology reports. Definite IA was defined as a positive culture and pathology showing septate hyphae. Probable IA was clinical disease and either a positive culture or histopathology. Disseminated IA was defined as involvement of two or more noncontiguous anatomic sites. RESULTS: We identified 33 cases of IA (28% disseminated) in 2,046 patients (attack rate = 1.6%) for an incidence of 4.8 cases per 1,000 patient-years (33 cases/6,813 pt-years). Both the attack and the incidence rates were significantly higher for lung transplant recipients vs. other transplant recipients: lung 12.8% (24 cases/188 patients) or 40.5 cases/1,000-pt year vs. heart 0.4% (3/686) or 1.4 per 1,000-pt year vs. liver 0.7% (3/439) or 2.1 per 1,000-pt year vs. renal 0.4% (3/733) or 1.2 per 1,000-pt year (P < 0.01). The incidence of IA was highest during the first year after transplantation for all categories, but cases occurred after the first year of transplantation only in lung transplant recipients. The attack rate of IA in lung transplant recipients was significantly lower after institution of routine Aspergillus prophylaxis (4.9% vs. 18.2%, P < 0.05). CONCLUSIONS: The highest incidence and attack rate of invasive aspergillosis among solid organ transplant recipients occurs in lung transplant recipients and supports the routine use of Aspergillus prophylaxis for at least one year after transplantation in this group. PMID- 12535263 TI - Ganciclovir-resistant cytomegalovirus encephalitis in a bone marrow transplant recipient. AB - A 20-year-old patient, who received a bone marrow transplant in order to treat metachromatic leukodystrophy (MLD), succumbed to cytomegalovirus (CMV) encephalitis. After CMV viremia developed, the patient received ganciclovir, but he was switched to foscarnet when ganciclovir resistance was suspected. Foscarnet was discontinued because of concern about its potential central nervous system toxicity. Autopsy samples of brain and cerebrospinal fluid contained CMV DNA with a UL97 mutation (M460V) known to confer ganciclovir resistance. No foscarnet resistance mutations were found. PMID- 12535264 TI - Disseminated Pseudallescheria boydii (Scedosporium apiospermum) infection in a renal transplant patient. AB - Transplant recipients receive a number of immunosuppressive medications that result in an increased risk of infection, including infections with microbes that are normally not pathogenic. We describe a patient with end-stage renal disease who underwent kidney transplantation. Six months postoperatively, he presented with a lesion on his ankle, multiple thigh nodules, and right testicular pain. Biopsy of the ankle lesion demonstrated Pseudallescheria boydii (Scedosporium apiospermum), a common environmental fungus. Following orchiectomy, multiple fungal elements were found that were initially described as Aspergillus species, but later identified as P. boydii. In addition, multiple brain abscesses were found on magnetic resonance imaging. Despite treatment with multiple antifungal medications, the patient died of cardiac dysrhythmia. Current diagnostic and therapeutic alternatives for P. boydii are reviewed. PMID- 12535265 TI - Breakthrough Scedosporium apiospermum (Pseudallescheria boydii) brain abscess during therapy for invasive pulmonary aspergillosis following high-risk allogeneic hematopoietic stem cell transplantation. Scedosporiasis and recent advances in antifungal therapy. AB - Systemic scedosporiasis due to the anamorph or asexual form Scedosporium apiospermum (Pseudallescheria boydii) has become an important cause of opportunistic mycosis, especially in patients undergoing high-risk hematopoietic stem cell transplantation. We report a case of rapidly progressive cerebellar hyalohyphomycosis due to Scedosporium apiospermum in an allogeneic marrow graft recipient receiving treatment for severe graft-versus-host disease. This fatal breakthrough intracranial abscess, due to amphotericin B-resistant (minimum inhibitory concentration > 16 micro g/ml) mold, developed during the course of systemic antifungal therapy given for multicentric pulmonary aspergillosis. Despite treatment with high-dose Abelcet (10 mg/kg daily), free amphotericin B was not detected in postmortem cerebellar tissue. A broad-spectrum triazole-based agent (voriconazole/UK-109, 496--Vfend), and a novel fungal cell wall inhibitor, an echinocandin/pneumocandin analog (caspofungin/MK-0991--Cancidas), which exhibit excellent in vitro activity against most clinical Pseudallescheria boydii Scedosporium apiospermum isolates, have recently become available in the United States and may provide much needed treatment options for patients at risk. PMID- 12535266 TI - An unusual abdominal mass in a renal transplant recipient. AB - Renal transplant recipients are at increased risk of malignancy and infection. We present the case of a 72-year-old-man with recurrent bladder carcinoma, abdominal aortic aneurysm repair, and end-stage renal failure due to renovascular disease. He received a cadaveric renal allograft into his left iliac fossa, was given cyclosporin A, azathioprine, and prednisolone triple therapy immunosuppression, and had no rejection episodes. He presented four years post-transplantation with a two-year history of intermittent sweats and fevers. Previous episodes had been investigated with no firm diagnosis made. This time he had right iliac fossa pain of three weeks' duration. Examination revealed a tender mass. Investigations showed unchanged graft function, but elevated inflammatory indices. Ultrasonography and computed tomography detailed an infiltrating mass associated with the sigmoid colon, which colonoscopy failed to visualise. At laparotomy a 6 cm tumor was removed, with adherent sigmoid colon and bladder dome. Macroscopically the mass was an abscess, and microscopy found acute and chronic inflammatory giant cells and fibrillary masses suggestive of actinomycosis, with no malignancy. The patient recovered uneventfully on antibiotics. At six months' follow-up, examination, inflammatory markers, and radiographic imaging showed no evidence of recurrence. Twelve months later the patient died of rupture of his proximal abdominal aorta. There was no evidence of recurrence at postmortem examination. We conclude with a brief review of actinomycosis in transplant recipients. PMID- 12535267 TI - Disseminated Nocardia asteroides and coinfection with Trichophyton rubrum in a renal transplant recipient. AB - Disseminated Nocardia infection has a high mortality and morbidity rate in solid organ transplant recipients, even when appropriate antibiotics are given. In this report a renal transplant recipient is described, who developed disseminated Nocardia asteroides infection with lung, eye, and brain involvement, in addition to coinfection with Trichophyton rubrum. PMID- 12535268 TI - Efficacy of Aspergillus galactomannan-directed preemptive therapy for the prevention of invasive aspergillosis in organ transplant recipients. PMID- 12535269 TI - Fusion and fission events in the endocytic pathway of Dictyostelium. AB - The endocytic pathway in Dictyostelium appears as a short circuit between endocytosis and exocytosis. Within the hour that elapses between internalization of nutrients and release of remnants, digestion by lysosomal enzymes occurs. Meanwhile, the maturing endosome undergoes a complex series of fusion and fission events, which change its character profoundly and which are far from being fully understood. This review attempts to order the dynamic events into a sequence of stages that is most consistent with present knowledge. PMID- 12535270 TI - The role of BEACH proteins in Dictyostelium. AB - The BEACH family of proteins is a novel group of proteins with diverse roles in eukaryotic cells. The identifying feature of these proteins is the BEACH domain named after the founding members of this family, the mouse beige and the human Chediak-Higashi syndrome proteins. Although all BEACH proteins share a similar structural organization, they appear to have very distinct cellular roles, ranging from lysosomal traffic to apoptosis and cytokinesis. Very little is currently known about the function of most of these proteins, few binding-partner proteins have been identified, and no molecular mechanism for any of these proteins has been discovered. Thus, it is important to establish good model systems for the study of these novel proteins. Dictyostelium contains six BEACH proteins that can be classified into four subclasses. Two of them, LvsA and LvsB, have clearly distinct roles in the cell. LvsA is localized on the contractile vacuole membrane and is essential for cytokinesis and osmoregulation. LvsB is most similar in sequence to the mammalian beige/Chediak-Higashi syndrome proteins and shares with them a common function in lysosomal trafficking. Structural and functional analysis of these proteins in Dictyostelium will help elucidate the function of this enigmatic novel family of proteins. PMID- 12535271 TI - Membrane traffic in virtual reality. PMID- 12535272 TI - Self-assembly is important for TIP47 function in mannose 6-phosphate receptor transport. AB - TIP47 (tail-interacting protein of 47 kDa) binds to the cytoplasmic domains of mannose 6-phosphate receptors and is required for their transport from endosomes to the trans-Golgi network in vitro and in living cells. TIP47 occurs in cytosol as an oligomer; it chromatographs with an apparent mass of approximately 300 kDa and displays an S-value of approximately 13. Recombinant TIP47 forms homo oligomers that are likely to represent hexamers, as determined by chemical cross linking. Removal of TIP47 residues 1-151 yields a protein that behaves as a monomer upon gel filtration, yet is fully capable of binding mannose 6-phosphate receptor cytoplasmic domains. The presence of an oligomerization domain in the N terminus of TIP47 was confirmed by expression of N-terminal residues 1-133 or 1 257 in mammalian cells. Co-expression of full-length TIP47 with either of these fragments led to the formation of higher-order aggregates of wild-type TIP47. Furthermore, the N-terminal domains expressed alone also occurred as oligomers. These studies reveal an N-terminal oligomerization domain in TIP47, and show that oligomerization is not required for TIP47 recognition of mannose 6-phosphate receptors. However, oligomerization is required for TIP47 stimulation of mannose 6-phosphate receptor transport from endosomes to the trans-Golgi in vivo. PMID- 12535273 TI - Arf regulates interaction of GGA with mannose-6-phosphate receptor. AB - The role of ADP-ribosylation factor (Arf) in Golgi associated, gamma-adaptin homologous, Arf-interacting protein (GGA)-mediated membrane traffic was examined. GGA is a clathrin adaptor protein that binds Arf through its GAT domain and the mannose-6-phosphate receptor through its VHS domain. The GAT and VHS domains interacted such that Arf and mannose-6-phosphate receptor binding to GGA were mutually exclusive. In vivo, GGA bound membranes through either Arf or mannose-6 phosphate receptor. However, mannose-6-phosphate receptor excluded Arf from GGA containing structures outside of the Golgi. These data are inconsistent with predictions based on the model for Arf's role in COPI veside coat function. We propose that Arf recruits GGA to a membrane and then, different from the current model, 'hands-off' GGA to mannose-6-phosphate receptor. GGA and mannose-6 phosphate receptor are then incorporated into a transport intermediate that excludes Arf. PMID- 12535274 TI - SopD2 is a novel type III secreted effector of Salmonella typhimurium that targets late endocytic compartments upon delivery into host cells. AB - Salmonella typhimuriumis a facultative intracellular pathogen that utilizes two type III secretion systems to deliver virulence proteins into host cells. These proteins, termed effectors, alter host cell function to allow invasion into and intracellular survival/replication within a vacuolar compartment. Here we describe SopD2, a novel member of the Salmonella translocated effector (STE) family, which share a conserved N-terminal type III secretion signal. Disruption of the sopD2 gene prolonged the survival of mice infected with a lethal dose of Salmonella typhimurium, demonstrating a significant role for this effector in pathogenesis. Expression of sopD2 was induced inside host cells and was dependent on functional ssrA/B and phoP/Q, two component regulatory systems. HA-tagged SopD2 was delivered into HeLa cells in a SPI-2-dependent manner and associated with both the Salmonella-containing vacuole and with swollen endosomes elsewhere in the cell. Subcellular fractionation confirmed that SopD2 was membrane associated in host cells, while the closely related effector SopD was localized to the cytosol. A SopD2 fusion to GFP associated with small tubular structures and large vesicles containing late endocytic markers, including Rab7. Surprisingly, expression of N-terminal amino acids 1-150 of SopD2 fused to GFP was sufficient to mediate both binding to late endosomes/lysosomes and swelling of these compartments. These findings demonstrate that the N-terminus of SopD2 is a bifunctional domain required for both type III secretion out of Salmonella as well as late endosome/lysosome targeting following translocation into host cells. PMID- 12535275 TI - Meeting report from the 55th Harden Conference: Dynamics of membrane traffic, 26 29th August 2002. PMID- 12535278 TI - Hachiro Tagami, MD: Japanese dermatologist in a global community. PMID- 12535279 TI - Mechanobiology of force transduction in dermal tissue. AB - BACKGROUND/AIMS: The influence of mechanical forces on skin has been examined since 1861 when Langer first reported the existence of lines of tension in cadaver skin. Internal tension in the dermis is not only passively transferred to the epidermis but also gives rise to active cell-extracellular matrix and cell cell mechanical interactions that may be an important part of the homeostatic processes that are involved in normal skin metabolism. The purpose of this review is to analyse how internal and external mechanical loads are applied at the macromolecular and cellular levels in the epidermis and dermis. METHODS: A review of the literature suggests that internal and external forces applied to dermal cells appear to be involved in mechanochemical transduction processes involving both cell-cell and cell-extra-cellular matrix (ECM) interactions. Internal forces present in dermis are the result of passive tension that is incorporated into the collagen fiber network during development. Active tension generated by fibroblasts involves specific interactions between cell membrane integrins and macromolecules found in the ECM, especially collagen fibrils. Forces appear to be transduced at the cell-ECM interface via re-arrangement of cytoskeletal elements, activation of stretch-induced changes in ion channels, cell contraction at adherens junctions, activation of cell membrane-associated secondary messenger pathways and through growth factor-like activities that influence cellular proliferation and protein synthesis. CONCLUSIONS: Internal and external mechanical loading appears to affect skin biology through mechanochemical transduction processes. Further studies are needed to understand how mechanical forces, energy storage and conversion of mechanical energy into changes in chemical potential of small and large macromolecules may occur and influence the metabolism of dermal cells. PMID- 12535280 TI - Quantitative description of human skin water dynamics by a disposition decomposition analysis (DDA) of trans-epidermal water loss and epidermal capacitance. AB - BACKGROUND: In vivo water assessment would greatly benefit from a dynamical approach since the evaluation of common related variables such as trans-epidermal water loss or "capacitance" measurements is always limited to instantaneous data. Mathematical modelling is still an attractive alternative already attempted with bi-exponential empirical models. A classical two-compartment interpretation of such models raises a number of questions about the underlying fundamentals, which can hardly be experimentally confirmed. However, in a system analysis sense, skin water dynamics may be approached as an ensemble of many factors, impossible to discretize, but conceptually grouped in terms of feasible properties of the system. The present paper explores the applicability of this strategy to the in vivo water dynamics assessment. METHODS: From the plastic occlusion stress test (POST) skin water balance is assessed by modelling trans-epidermal water loss (TEWL) and "capacitance" data obtained at skin's surface. With system analysis (disposition-decomposition analysis) the distribution function, H(t), modelled as a sum of exponential terms, covers only the distribution characteristics of water molecules traversing the skin. This may correspond macroscopically to the experimental data accessed by "corneometry". Separately, the hyperbolic elimination function Q(TEWL) helps to characterise the dynamic aspects of water influx through the skin. DISCUSSION AND CONCLUSION: In the observable range there seems to be a linear relationship between the net amount of water lost at the surface by evaporation, and the capability of the system to replenish that loss. This may be a specific characteristic of the system related to what may be described as the skin's "intrinsic hydration capacity" (IHC) a new functional parameter only identified by this strategy. These new quantitative tools are expected to find different applicabilities (from the in vivo skin characterisation to efficacy testing) contributing to disclose the dynamical nature of the skin water balance process. PMID- 12535281 TI - Water, salts and skin barrier of normal skin. AB - BACKGROUND: We recently reported that open application of seawater for 20 min ameliorated experimental irritant contact dermatitis induced by sodium lauryl sulphate (SLS) cumulative irritation. The efficacy was overall contributed by 500 mm of sodium chloride (NaCI) and 10 mm of potassium chloride (KCl), which are consistent with the each concentration in seawater. Although the usefulness of mineral water with 500 mm NaCl and 10 mm KCl to treat atopic dermatitis (AD) or irritated skin was considered, seawater or its components would induce a feel of stinging in patients with AD. Furthermore, 20 min application was thought to be too long to use everyday as a treatment. OBJECTIVE: We report the effects of 3 types of mineral water with NaCl and KCl to check the further efficacy with lesser stinging by 2 min application. RESULTS: A mineral water with 250 mm NaCl and 50 mm KCl was the most effective water to prevent disruption of skin barrier and stratum corneum water content after cumulating irritation by SLS. Moreover, improvement of skin dryness and pruritus were shown 2 weeks after the application of the mineral water to a 6-year-old boy with atopic dermatitis. CONCLUSION: Our results suggested the possible usefulness of the mineral water with 250 mm NaCl and 50 mm KCl as the therapy of atopic dermatitis of other chronic dermatitis. Although the mineral water would not cure those skin diseases, it could be an adjunctive therapy. Further controlled clinical trials with evaluation by TEWL and capacitance are required to declare the efficacy of the mineral water in the treatment of patients with AD or other chronic dermatitis. PMID- 12535282 TI - Impact of stress of marital dissolution on skin barrier recovery: tape stripping and measurement of trans-epidermal water loss (TEWL). AB - BACKGROUND: Psychological stress of marital disruption is associated with significant increases in a variety of psychological and physical disorders. The effect of stress on the immune system is well documented and skin disorders have been reported to exacerbate during stressful situations. This study was designed to observe the effects of stress on skin barrier strength and recovery. Twenty eight healthy females age 21-45 who were in the process of marital separation were tested for skin barrier strength and recovery. The panel was chosen on the basis of the intensity of self perceived stress. The control group was an age matched group of self perceived 'happy' subjects. Servomed evaporimeter was used to measure trans-epidermal water loss (TEWL) from cheek area of the face, before and after removing stratum corneum layers with tape strippings. Skin barrier strength was defined as the number of tape strippings required to disrupt skin barrier, which is a TEWL of 18 g/m2/h or more. Barrier recovery was denoted by the level of TEWL, 3 h and 24 h after barrier disruption. RESULTS: There was no correlation between the degree of stress and barrier strength. However, individuals with high stress recovered slower than the individuals with low stress after 3 h (R = 0.64) and 24 h (R = 0.74). CONCLUSIONS: Psychological stress of marital dissolution does not appear to change skin barrier strength but has a negative impact on skin barrier recovery. PMID- 12535283 TI - Radiation-induced skin fibrosis after treatment of breast cancer: profilometric analysis. AB - BACKGROUND/AIM: Treatment of breast cancer involves an association of conservative surgery and radiotherapy. This implies various cutaneous complications, well known for their clinical and histological aspects. Little data are available concerning modifications of the cutaneous microrelief after radiotherapy. We have done a profilometric analysis of the skin of breast cancer patients treated with surgery and radiotherapy. The results obtained on the irradiated breast have been compared with the ones of the controlateral normal breast. METHODS: Twenty women, 43-55 year old were enrolled in the study 6-16 months after the end of a treatment associating conservative surgery and radiotherapy for breast cancer. Imprints using a silicone rubber material were performed over symmetrical areas of the treated breast and the controlateral one. All measures were performed over the upper-medial quadrant of each breast. The imprints are then blindly analysed using an image analyser. The following parameters were measured: RA (average roughness of the skin), RZ (average of the furrows depth), RS (residual length), Rn (number of furrows) and AR (space between the furrows). RESULTS: The comparison of the imprints shows important modifications of the microrelief after irradiation. We observe a slight increase of the skin roughness (RA: 26, 39 +/- 2, 58 versus 21, 84 +/- 1, 59), a significantly increase of the furrows depth (RZ: 121, 66 +/- 10, 46 versus 101, 26 +/- 6, 99) along with an increase of the residual length (RS: 580, 60 +/- 60, 60 versus 450, 46 +/- 48, 43). The number of furrows has not significantly decreased but the space between the furrows has increased on the irradiated breast. CONCLUSION: This study shows a breakdown effect of irradiation on the skin in correlation with the fibrosis inducing by ionising radiation. The imprints modifications are clearly different from those usually observed in the ageing process. Our results can be a basis for studying the effects of treatments on cutaneous complications linked to the radiation-induced fibrosis. PMID- 12535284 TI - The use of different concentrations of betaine as a reducing irritation agent in soaps monitored visually and non-invasively. AB - BACKGROUND/AIMS: Products containing detergents can damage the skin and give rise to irritant contact dermatitis. Therefore, attempts have been made to find less irritating detergents as well as substances decreasing undesired side-effects of detergents, and a novel approach is offered by betaine. The aim of the study has been to determine the irritating properties of some liquid soaps for personal hygiene and to map the effect of different concentrations of betaine using electrical impedance, trans-epidermal water loss and visual inspection. METHODS: Twenty-eight healthy subjects were patch tested with different commercial soaps with and without betaine and sodium lauryl sulphate on both volar forearms for 24 h. A site with distilled water and an unoccluded area were used as references. Responses of the skin reactions were evaluated by visual inspection and by measuring trans-epidermal water loss and electrical impedance before application and 24 h after removal of the chambers. RESULTS/CONCLUSIONS: Significant skin reactions were found for all soaps tested but the soaps containing betaine were the least irritating. However, the skin irritation did not decrease with increasing concentrations of betaine in the tested range. On the whole the differences between the products were not large. The non-invasive methods used were more sensitive than visual assessment for evaluation of invisible or barely visible skin responses. PMID- 12535285 TI - The ability of betaine to reduce the irritating effects of detergents assessed visually, histologically and by bioengineering methods. AB - BACKGROUND/AIMS: A novel approach for reducing the undesired irritating properties of detergents on skin might be offered by betaine, which is a natural product derived from the sugar beet. The aim of the study was to explore the ability of betaine to reduce the irritating effects of two surfactants, sodium lauryl sulphate (SLS) and cocoamidopropylbetaine (CAPB). For evaluation of changes in skin reactions visual scoring, electrical impedance, transepidermal water loss and histology were used. METHODS: Twenty-one healthy subjects were patch tested for 24 h with SLS and CAPB alone and together with betaine, betaine alone, and the two controls distilled water and an unoccluded test site on both volar forearms. Responses were evaluated by measuring electrical impedance and transepidermal water loss before exposure and 24 h after the removal of the test substances, and also by visual inspection and histology. The electrical impedance device enables measurements at 31 frequencies and relevant information was extracted from the spectra using four indices. RESULTS: CAPB was found to be less irritating than SLS. The used detergents gave rise to distinctive impedance patterns also reflected by different types of histopathological skin responses. After the adding of betaine, the irritant reaction decreased for both detergents. CONCLUSIONS: Betaine is a promising ingredient to reduce the side effects of detergents and electrical impedance is a suitable tool both to quantify the degree of irritation as well as to differentiate between various types of reactions. PMID- 12535286 TI - Differences among moisturizers in affecting skin susceptibility to hexyl nicotinate, measured as time to increase skin blood flow. AB - BACKGROUND/AIMS: A wide range of branded and generic moisturizers is frequently used for the prevention and treatment of dry skin. The influence of moisturizers on the skin permeability is pertinent to the understanding of their therapeutic efficacy. The aim of the present study was to compare the effect of two moisturizers on the skin permeability barrier, assessed as skin reactivity to a vasodilating substance. METHODS: The study was parallel, randomized and double blind on 53 healthy volunteers. One of the creams contained 5% urea, whereas the other contained no humectant but had a high lipid content. The participants were instructed to apply the cream twice daily for three weeks on the volar aspect of one of their forearms. The skin was then exposed to hexyl nicotinate, which induces vasodilation. The time-course and magnitude of the microvascular changes in the two skin areas were monitored with a non-invasive optical technique (laser Doppler flowmetry) with two measuring probes. RESULTS: The lag-time between application and initial response was significantly longer for the urea-treated site compared with the other cream. Furthermore, the time for maximum response was shorter for the lipid-rich cream than for its placebo. CONCLUSION: The study shows differences in action between moisturizers, which may influence the skin susceptibility to other irritants and allergens in the environment. PMID- 12535287 TI - Validation and clinical relevance of a novel scalp coverage scoring method. AB - BACKGROUND/AIMS: Distinctive patterns of defective scalp skin coverage or alopecia have been identified by clinicians but do not help measuring the dynamics of hair changes over time. Hence, we conceived the scalp coverage scoring (SCS) method that integrates cumulative changes of hair growth or loss. In this paper, we report the results of validation studies demonstrating that the SCS can be applied in real time in the clinic or on global photographs for evaluating the severity of androgenetic alopecia (AGA) of male subjects and for monitoring changes over time. METHODS: Different panels totalling 38 different male subjects with AGA were classified according to a modified Norwood-Hamilton scale and were evaluated with the SCS system (real time or global photographs). The contrast-enhanced phototrichogram method (CE-PTG), a refined non-invasive and validated analytical technique was used for calibration purposes. RESULTS: The real time SCS was correlated with SCS on global photographs. The inter- and intra observer variation of SCS obtained on global photographs was less than 3%. The real time SCS was negatively correlated with the clinical staging of male pattern baldness according to the modified Norwood-Hamilton scale, confirming less scalp coverage in more severly affected subjects. In 12 male subjects with AGA, after SCS evaluation of the top of the head, a 1-cm2 central target site was explored with CE-PTG. There was a significant correlation of real time SCS with the percentage of thick anagen hair (r = 0.53; P < 0.08) and with the density of thin hair (r = -0.68; P < 0.02). Changes over time as documented on global photographs (18 subjects at baseline and after 12 and 60 months) were rated by a panel of independent experts (paired photographs) who were not blinded as to time. Randomised individual photographs were also examined one at a time for SCS. This procedure documents data obtained without control of baseline photographs and establishes blinding as to time. The changes calculated from SCS (after-baseline) were correlated (r = 0.9; P < 0.0001) with the rating of the panel of experts. CONCLUSIONS: When minimum technical precautions are taken prior to the scoring session, the main points of interest of the SCS method are: Non-invasive quantitative and reproducible technique. Measures dynamics of scalp hair growth and loss. Easily applied in the hair clinic. PMID- 12535288 TI - Analysis of skin line pattern for lesion classification. AB - BACKGROUND/PURPOSE: It has been observed that skin patterning tends to be disrupted by malignant but not by benign skin lesions. This suggests that measurements of skin pattern disruption on simply captured white light optical skin images could be a useful contribution to a diagnostic feature set. Previous work using a measurement of line strength by a consistent high-value profiling technique followed by local variance measurement or a region agglomerative classifier to measure skin line pattern disruption was extremely promising but computationally intensive, suggesting that the idea of measuring skin pattern disruption was useful but a simpler method was required. METHODS: The skin pattern was extracted by high-pass filtration and enhanced by adaptive anisotropic (spatial variant) filtering which smoothes along skin lines but not across them. The skin line main direction and direction variance were estimated using a local image gradient matrix and the difference of these measures across the lesion image boundary was used as a lesion classifier. RESULTS: A set of images of malignant melanoma and benign naevi were processed as above and the scatter plot of results in a two-dimensional feature (line direction and line variation difference) space showed excellent separation of benign and malignant lesions. An ROC plot enclosed an area of 0.88. CONCLUSIONS: The experimental results showed that the local line direction and the local line variation were promising features for distinguishing malignant melanoma from benign lesion and the methods used were effective and computationally low-cost. PMID- 12535289 TI - Reproducibility of repeated measurements on post-burn scars with Dermascan C. AB - The present study investigated the reproducibility of measurements with the Dermascan C on post-burn scars. Intra- and inter-rater reproducibility and test retest reliability with a one week time lapse were examined on 40 post-burn scars of 6 volunteering burn patients by means of intra-class- correlation coefficients (ICC) and standard error of measurements (SEM). Results showed good to excellent values for ICC in all the three conditions. On the basis of these results, we concluded that the instrument provides reliable information and can be used in comparative clinical trials that examine therapeutic strategies on healing scars. PMID- 12535290 TI - The ADEMEX study and its implications for peritoneal dialysis adequacy. AB - In an attempt to improve patient outcomes in peritoneal dialysis (PD), national organizations (such as the National Kidney Foundation Dialysis Outcomes Quality Initiative [NKF-DOQI] process) have formulated clinical practice guidelines based on clinical evidence available at the time of development. For "adequacy" of PD it was acknowledged that there was no prospective randomized interventional clinical trial that evaluated the effect of an increase in peritoneal clearance on outcome. The ADEMEX study is the first such study designed to do this. It was well done and adequately powered for the primary analysis. The study findings indicate that over the range of solute clearances studied, an increase in peritoneal clearance is not associated with an incremental improvement in patient outcome. However, it is noted that the cause of dropout was different between groups, with more dropout for "uremia" in the control group. There are also some limitations in the generalizability of the results. First, the exclusion criteria were likely to exclude patients who were small in body size or were high transporters, patients with the highest relative risk of death. Second, although there was an increase in small solute clearance between control and intervention groups, there was not likely to be an increase in clearance of other potential uremic solutes such as middle molecules. Third, the study did not examine outcomes for patients on cycler therapy. Nevertheless it was a provocative, well run clinical study which does have implications for clinical practice. It confirms that one prescription does not fit all patients, that many patients below current NKF-DOQI targets for small solute clearance are likely to be adequately dialyzed, and provides evidence-based clinical information for national societies to consider when preparing for the next revision of their guidelines. PMID- 12535291 TI - Treating interdialytic hyperkalemia with fludrocortisone. AB - Hyperkalemia is a frequent and dangerous problem in dialysis patients. Many factors contribute to potentially life-threatening potassium elevation and most remedies used to treat hyperkalemia are handicapped by the consequences of the separate pools of intra- and extracellular potassium. Besides the kidney, the colon has the ability to excrete potassium, which can help lower total body potassium. Several prior authors have addressed the colon's ability to up regulate potassium secretion, including the effect of aldosterone on fecal potassium content. Potentially dangerous intradialytic maneuvers to lower potassium levels may be avoidable with the use of the mineralocorticoid agonist fludrocortisone. PMID- 12535292 TI - How will the results of the HEMO study impact dialysis practice? PMID- 12535297 TI - Approach to patients with "complex" hemodialysis access problems. AB - The approach to patients with "complex" permanent hemodialysis (HD) access problems remains poorly defined. The purpose of this review is to outline our current algorithm for patients presenting for dialysis access and to address the management of specific problems that complicate this objective of establishing permanent access. The key components of the algorithm include noninvasive imaging in the diagnostic vascular laboratory to determine all the possible access configurations and invasive imaging with both venography and arteriography to confirm the optimal choice. The specific access-related problems addressed include inadequate ipsilateral vein, inadequate arterial inflow, central vein stenosis/occlusion, multiple previous access failures, and obesity. Despite the label of "complex" access problems, it is possible to construct native arteriovenous fistulas (AVFs) in almost all patients presenting for access using the standard principles of vascular surgery that are based on establishing adequate arterial inflow, adequate venous outflow, and selecting an appropriate conduit. PMID- 12535298 TI - Infection with antimicrobial-resistant microorganisms in dialysis patients. AB - The prevalence of antimicrobial-resistant microorganisms in various health care settings, including outpatient dialysis facilities, has increased dramatically in the last decade. Antimicrobial use and patient-to-patient transmission of resistant strains are the two main factors that have contributed to this rapid increase. Methicillin-resistant Staphylococcus aureus (MRSA) and coagulase negative staphylococci are commonly isolated as a cause of hemodialysis (HD) catheter-related bacteremia and peritoneal dialysis (PD)-related catheter infection and peritonitis. The widespread use of vancomycin in dialysis patients is of concern because of an increase in the prevalence of vancomycin-resistant enterococci (VRE) in dialysis patients. Staphylococci with reduced sensitivity to vancomycin have also appeared in dialysis patients. A more recent problem is the appearance of S. aureus isolates with a high degree of resistance to the topical antimicrobial agent mupirocin. This has been seen in PD patients who have received prophylactic application of mupirocin at the peritoneal catheter exit site. Appropriate antimicrobial use will help protect the efficacy of currently used antibiotics, such as vancomycin. Published guidelines for use of vancomycin should be followed. New antimicrobials such as linezolid and quinupristin/dalfopristin have activity against VRE and MRSA, but resistance to these agents has already occurred. Preventing transmission of antimicrobial resistant microorganisms in health care settings, including outpatient dialysis facilities, is important in limiting the spread of these resistant organisms. PMID- 12535299 TI - Tuberculosis and chronic renal disease. AB - There is an increased risk (6.9- to 52.5-fold) of tuberculosis (TB) in patients with chronic renal failure and on dialysis as compared to the general population. The symptomatology in renal patients is often insidious and nonspecific, mimicking uremic symptoms, whereas the localization is often extrapulmonary (most frequently tuberculous lymphadenitis and peritonitis). Tuberculous peritonitis makes up a large part (37%) of the total number of TB cases in continuous ambulatory peritoneal dialysis (CAPD) patients. The prognosis is very much dependent on early diagnosis and treatment. Renal physicians should be aware of the unusual presentation and localization, and include TB in the differential diagnosis of any patient having nonspecific symptoms like anorexia, fever, and weight loss. All efforts should then be made (including invasive investigations) to reach an early diagnosis, a major determinant of the outcome. However, if this is not possible or the result is negative and the diagnosis remains strongly suspected, an empirical trial with anti-TB medication is justified, especially in endemic areas. In view of the increased prevalence of the disease in the dialysis population, TB prophylaxis is recommended in those patients with a positive tuberculin (Mantoux) skin test and radiographs suggestive of old TB. PMID- 12535300 TI - The effect of chronic renal failure on hepatic drug metabolism and drug disposition. AB - There is abundant evidence that chronic renal failure (CRF) and end-stage renal disease (ESRD) alter drug disposition by affecting protein and tissue binding and reducing systemic clearance of renally cleared drugs. What is not fully appreciated is that CRF can significantly reduce nonrenal clearance and alter the bioavailability of drugs predominantly metabolized by the liver. Animal studies in CRF have shown a major down-regulation (40-85%) of hepatic cytochrome P-450 metabolism involving specific isozymes. Phase II reactions such as acetylation and glucuronidation are also involved, with some isozymes showing induction and others inhibition. Hepatic enzymes exhibiting genetic polymorphisms such as N acetyl-transferase-2 (NAT-2), which is responsible for the rapid and slow acetylator phenotypes, have been shown to be inhibited by ESRD and reversed by transplantation. There is some evidence pointing to the possibility of inhibitory factors circulating in the serum in ESRD patients which may be dialyzable. This review includes all significant animal and clinical studies using the search terms "chronic renal failure,""cytochrome P-450," and "liver metabolism" over the past 10 years obtained from the National Library of Medicine MEDLINE database, including relevant articles back to 1969. PMID- 12535301 TI - Cardiopulmonary resuscitation and dialysis: outcome and patients' views. AB - Cardiopulmonary resuscitation (CPR) was initially described as an intervention to be used in otherwise healthy individuals suffering acute cardiorespiratory arrest. Over the years, CPR has been extended to all hospitalized patients unless specific orders not to resuscitate have been written with the informed consent of the patient and/or surrogate. The 14-15% survival to hospital discharge reported for in-hospital CPR has not changed over the past three decades. Compared with other diseases, chronic kidney disease reduces long-term survival (more than 6 months) following CPR, and the functional status of the few who survive is often quite poor. Nevertheless, most dialysis patients want to be resuscitated. Unfortunately television shows portraying resuscitation imply that survival after CPR is much more common than it really is. Such misinformation contributes to the overwhelming choice for CPR despite the dismal prognosis. Dialysis unit staff need to educate patients and families about the expected success and complications of CPR as part of the advance care planning process that should now be routine. PMID- 12535302 TI - The pathophysiology of immune-mediated heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is an important side effect of heparin therapy associated with significant morbidity and mortality if unrecognized. The platelet count typically falls below 150,000/ micro l 5-14 days after heparin is started. Thrombosis is the major clinical complication. The diagnosis is confirmed with a variety of functional and antigenic assays. Heparin binds to PF4, resulting in a conformational change in the molecule that exposes neo epitopes that act as immunogens. Antibodies form against the heparin-PF4 complex, the major target antigen. The IgG-heparin-PF4 immune complex binds either via its Fab domain to the platelet surface or via its Fc domain to the FcgammaIIA receptor on the surface of the platelet, resulting in further platelet activation. Continued release of PF4 from activated platelets leads to increasing PF4-heparin complex formation, and a self-propagating cycle of platelet consumption and generation of procoagulant platelet-derived microparticles. Other procoagulant effects of the HIT antibody include endothelial cell damage, stimulation of platelet-leukocyte aggregates, and release of tissue factor from monocytes. PMID- 12535303 TI - Alternative methods of anticoagulation for dialysis-dependent patients with heparin-induced thrombocytopenia. AB - Dialysis patients who are continually exposed to heparin are at risk for heparin induced thrombocytopenia (HIT). Heparin-induced antibodies have been reported to occur in 0-12% of hemodialysis (HD) patients. The diagnosis or suspicion of HIT in this patient population requires careful confirmation of the diagnosis and substitution of heparin with an alternate anticoagulant for dialysis. Alternate agents such as the direct thrombin inhibitors (hirudin and argatroban) are available, but careful dosing and monitoring of the anticoagulant effect are required. Despite careful dosing, hemorrhagic complications have occurred with these agents. Unfortunately there are limited options for treatment of hemorrhagic complications and no specific antidotes are available for the direct thrombin inhibitors. In this report the currently available alternatives to heparin for dialysis, including dosing and monitoring recommendations, are reviewed. PMID- 12535304 TI - Does uremia protect against the demyelination associated with correction of hyponatremia during hemodialysis? A case report and literature review. AB - Rapid correction of chronic hyponatremia is known to cause demyelination syndromes, which are attributed to the rapid shift of water out of the brain. In uremic patients with hyponatremia, depending on the dialysate sodium concentration and delivered Kt/V, serum sodium levels may be rapidly corrected inadvertently during the hemodialysis (HD) session. It is not known whether uremic patients are as susceptible to the development of demyelination as patients with normal renal function. Since urea diffuses slowly across the blood brain barrier, it can act as an effective osmole between plasma and the brain if levels are changed abruptly. During HD, blood urea levels drop suddenly and significantly and cerebral edema may develop (dialysis disequilibrium syndrome). This effect may counteract the fluid shift out of the brain during correction of hyponatremia. Therefore, theoretically, uremic patients may be less prone to develop demyelination. We present a patient with renal failure whose hyponatremia was corrected rapidly during HD to illustrate the potential problem. The patient tolerated rapid correction of hyponatremia without sustaining any neurologic damage. We performed a literature search looking for similar case reports and reviewed the scientific evidence behind the above hypothesis. PMID- 12535305 TI - Peritoneal dialysis catheter replacement: "save the patient and not the catheter". AB - A 63-year-old man on ambulatory peritoneal dialysis (APD) developed peritonitis with Enterococcus faecalis, treated with 2 g intraperitoneal (IP) vancomycin, with rapid clearing of the effluent white blood cells, but persistence of positive cultures. Vancomycin was redosed on day 8 (2 g IP), and then weekly. Gentamicin 140 mg IP loading dose, followed by 40 mg IP once a day was added after cultures were still positive at 2 weeks. The two drugs were continued for six additional weeks. Although the patient was asymptomatic, the effluent cultures continued to grow E. faecalis and the catheter was replaced 2 months after the onset of peritonitis. There was no evidence of either tunnel infection or intra-abdominal abscess. Refractory peritonitis is defined as continuation of peritonitis after 5 days of appropriate therapy. This patient had persistently positive cultures but quickly became asymptomatic and signs of inflammation resolved readily. The most likely etiology appears to have been colonization of the slime layer of the intra-abdominal portion of the catheter with the organism. The vancomycin dosing schedule may have played a role in the persistently positive cultures. A recent pharmacokinetic study suggested that patients on APD require 35 mg/kg IP of vancomycin as a loading dose, followed by 15 mg/kg IP once a day, given in a long day exchange. Simultaneous placement and removal as a single procedure was successful in this patient and can be done safely in patients whose effluent white blood cell count (WBC) is less than 100/mm3. Most patients can then be subsequently managed by doing supine dialysis (using a cycler) with decreased exchange volumes and a dry abdomen until healing occurs (usually 1-2 weeks). In this way hemodialysis can be avoided. By minimizing the effect on the patient's lifestyle, the patient is more likely to agree to a catheter exchange. PMID- 12535306 TI - Black holes and ring nebulae. PMID- 12535307 TI - Insomnia on dialysis nights. PMID- 12535308 TI - The risk of preoperative hyperkalemia. PMID- 12535309 TI - Angiotensin receptor blockers and epoetin resistance. PMID- 12535311 TI - Muscle strength, disability and mortality. AB - The aims of this review are to address (1) the role of muscle strength in the disablement process and (2) muscle strength as a predictor of length of life using data from prospective studies. Functional limitations, such as slow walking speed, predispose older people to disabilities. How much strength is needed for daily motor tasks such as walking varies according to other impairments present. For example, when postural balance is good, only minimum amount of strength is needed for walking. However, in the presence of balance impairment, having good level of strength may help to compensate for the deficit. Having strength well above the required level indicates reserve capacity. It was studied using data from the Honolulu Heart Program launched in 1965 among 8006 men initially aged 45 68 years, whether reserve of strength would be protective of development of future disability. All men with documented diseases at baseline were excluded from the analyses. Those men who were in the lowest third of the distribution of grip strength at baseline were at two to three times greater risk of developing disabilities assessed 25 years later compared to the highest third. It is possible that before they reach the disability level, those with greater reserve of strength may afford to lose more strength, for example following bed rest and inactivity associated with an illness. Midlife grip strength was also found to predict long-term total mortality: those with poorer strength at baseline were more likely to die over the follow-up period of 30 years. The association between muscle strength and disability is largely explained by biomechanical mechanisms. However, the mechanism explaining the association between muscle strength and mortality risk still remains to be explored. PMID- 12535312 TI - The influence of genetic factors on physical functioning and exercise in second half of life. AB - During the past decades, a number of studies of families and twins in particular have assessed the relative contribution of genetic and environmental factors to traits reflecting various aspects of physical functioning: maximal O2 uptake, muscular endurance, muscular strength, muscle cross sectional area, flexibility, and trainability. Although the estimate of the size of the genetic component differs between the various studies, they point towards a moderate to substantial genetic influence on these phenotypes. Most of the studies have used only young and healthy study subjects, although in recent years phenotypes of particular importance to the elderly and the oldest-old (e.g., activities-of-daily living abilities) have also been shown to have substantial genetic component. Furthermore, behavioural studies have also revealed a genetic contribution to the disposition to level of leisure time physical activity. At present, there is still a few association studies on specific genetic variants, and the results have either been inconsistent or failed to show an association with physical functioning. Therefore, the mechanisms through which the genetic influence is expressed, is still an enigma. Here, we summarise the evidence currently available for a genetic influence on physical functioning and disposition to leisure time physical activity with a focus on recent Danish twin data. PMID- 12535313 TI - Muscle performance, sex hormones and training in peri-menopausal and post menopausal women. AB - Age-related deterioration in muscle performance is one of the major reasons for decreased functional capacity and disability in older people. In women, impaired muscle performance has already been observed during peri-menopause in concert with rapid and dramatic decrease in ovarian hormone production. This observation suggests that female sex steroids may have an important role among other agents in regulating muscle performance in middle-aged and older women. Previous experimental studies have shown that hormone replacement therapy (HRT) and intensive physical training have positive effects on muscle force and explosive power in healthy post-menopausal women. Hormone replacement therapy in combination with physical training may exert even greater gains in muscle performance than HRT and training alone. Despite the significant mean increase in muscle force and power by HRT and/or training, a considerable variability in the individual responses is observed. The mechanism, by which female sex steroids act on muscle performance, is still unclear. Therefore, more research is needed in order to explore all the pathways, by which these steroids could act on skeletal muscle in peri-menopausal and post-menopausal women. PMID- 12535314 TI - Effects of ageing and human whole body and muscle protein turnover. AB - Prevalence of sarcopenia is up to 60% of those individuals over 80 years of age and is associated with increased disability. The causes behind the age-related loss of muscle are difficult to discern. Measurements of protein synthesis/breakdown and net protein balance are important, and further methodological development is warranted. Whole body protein turnover is changed only little - if at all - with ageing, when corrected for fat free mass of the individuals. Discrepancies in reports are often related to inconsistent recordings of energy intake especially protein and variation in subject, gender and physical activity level. Ageing is associated with reduced sensitivity toward amino acids, increased first pass uptake in a splanchnic region and a reduced postprandial stimulation of protein synthesis. Physical activity and amino acids are additive in effect also in elderly individuals, and timing of training and protein intake is crucial, in that early intake of amino acids is advantageous with regards to stimulation of protein synthesis. PMID- 12535315 TI - Ageing and local growth factors in muscle. AB - Muscle responds to mechanical overload by increasing its size. In contrast, as a muscle gets older it atrophies. The mechanisms regulating these differing responses are not fully understood. Animal studies have shown that older muscles are less well able to repair following contraction-induced injury than young muscles. It is becoming clear that local growth factors produced within the muscle may play important roles in both repair, adaptation and ageing. The growth hormone/insulin like growth factor 1 (GH/IGF-I) axis is important during growth and development, but circulating levels of these hormones decline in later life. However, many tissues including muscle, produce IGF-I for autocrine and paracrine actions. Genetic manipulation of IGF-I in muscle has shown that it has considerable anabolic affects on muscle both in young and old animals. Insulin like growth factor 1 exists in multiple isoforms and one isoform, which differs from the systemic or liver type (IGF-IEa), appears to be particularly sensitive to mechanical signals and to muscle damage. This isoform (IGF-IEc) has been termed mechano growth factor (MGF). The anabolic actions of IGF-I and MGF are through stimulating protein synthesis and by playing a role in the activation, proliferation and differentiation of satellite cells. These effects are discussed in relation to human studies of muscle adaptation to strength training in older people who seem to retain an ability to increase muscle mass and strength through this type of exercise. PMID- 12535316 TI - Muscle fibre type adaptation in the elderly human muscle. AB - This short review discusses changes in the fibre type distribution, myosin heavy chain isoform composition and histological appearance of the very elderly human skeletal muscle. Point of origin of the discussion comes from data that we have obtained from muscle biopsies from the vastus lateralis muscle of a group of frail very elderly subjects (age: 88 +/- 3 years, range 85-97). Myosin heavy chain composition of muscle homogenates and single fibres, fibre type distribution, fibre size and capillary density were examined and compared with muscle biopsies from the young vastus lateralis muscle. Histological preparations of the muscle biopsies from our elderly subjects showed extended "grouping" (Nygaard & Sanchez, Anat Rec 1992: 202: 451-459) of the fibre types as well as significant changes in the appearance and size of the individual muscle fibres. On average, the fibre type composition of our very elderly subjects do not seem to be different to what is observed in a corresponding young group when examined with ATPase histochemistry. Likewise, the MHC composition of the muscle homogenates is comparable to what is observed in young subjects. Nevertheless, a detailed examination of the MHC composition of single fibres from the old subjects revealed that the most prominent phenotype was fibres co-expressing MHC I and MHC IIA. This is very different from what is observed in the young muscle. Detailed investigation of longitudinally cut fibres indicated that some fibres in the very old muscle, in contrast to the young muscle, switch fibre type along the length of the fibre or contain areas or nuclear domains in which the MHC expression is different from the remaining part of the fibre. PMID- 12535317 TI - Satellite cells and training in the elderly. AB - In the present review, we describe the effects of ageing on human muscle fibres, underlining that each human muscle is unique, meaning that the phenotype becomes specifically changed upon ageing in different muscles, and that the satellite cells are key cells in the regeneration and growth of muscle fibres. Satellite cells are closely associated with muscle fibres, located outside the muscle fibre sarcolemma but beneath the basement lamina. They are quiescent cells, which become activated by stimulation, like muscle fibre injury or increased muscle tension, start replicating and are responsible for the repair of injured muscle fibres and the growth of muscle fibres. The degree of replication is governed by the telomeric clock, which is affected upon excessive bouts of degeneration and regeneration as in muscular dystrophies. The telomeric clock, as in dystrophies, does not seem to be a limiting factor in ageing of human muscle. The number of satellite cells, although reduced in number in aged human muscles, has enough number of cell divisions left to ensure repair throughout the human life span. We propose that an active life, with sufficient general muscular activity, should be recommended to reduce the impairment of skeletal muscle function upon ageing. PMID- 12535318 TI - Possible beneficial role of exercise in modulating low-grade inflammation in the elderly. AB - Aging is associated with increased levels of tumor necrosis factor-alpha (TNF) and interleukin (IL)-6. These two cytokines are tightly linked in that TNF induces production of IL-6, which again inhibits TNF gene expression. In epidemiological studies, both cytokines have been associated with obesity, insulin resistance and atherosclerosis. However, based on basal studies, we suggest that TNF (and not IL-6) is the driver behind insulin resistancy. Thus, it is possible that selective enhancement of the IL-6 level may inhibit TNF-induced insulin resistance. Muscle contractions induce production and release of IL-6, but not TNF, into the circulation, in both young and elderly humans. We suggest that muscle-derived IL-6 contributes to mediate the beneficial metabolic effects of exercise and may contribute to inhibit TNF-production and thereby insulin resistance. PMID- 12535319 TI - Physical activity in an ageing population. AB - This review over previously published articles, describes physical activity changes during ageing from age 50-85 in the 1914-cohort in Glostrup, Denmark, and analyses the association between physical activity and mortality, myocardial infarction, hip fractures and functional ability. The 1914-cohort in Glostrup was examined at age 50 in 1964 and re-examined 1974, 1984, 1989, 1994 and 1999. Some analyses were based on pooled data (CCPPS: Copenhagen Centre for Prospective Population Studies) from three longitudinal population studies conducted in Copenhagen, Denmark: The Copenhagen County Centre for Preventive Medicine in Glostrup, The Copenhagen City Heart Study and The Copenhagen Male Study. Physical activity of work and leisure time was classified into four levels based on questions originally constructed by Saltin and Grimby. MAIN OUTCOME MEASURES: all cause mortality, fatal and non-fatal myocardial infarction, hip fracture and functional ability (Avlund mobility index of dependency). The lifetime risk from a sedentary leisure time was analysed in multivariate regression analyses controlling for covariates describing gender, age, life style, education and chronic diseases. Less than one third of the population had a sedentary leisure time. Physical activity definitely influenced health and quality of life over the life course in a positive way: compared to the group of inactive men and women, the mortality was about 60%, the incidence of myocardial infarction 70% and the incidence of hip fractures 75% in the moderate active groups. On top the physically active persons gained independency in activities of daily living. PMID- 12535320 TI - Effects of training on functional performance in 65, 75 and 85 year-old women: experiences deriving from community based studies in Odense, Denmark. AB - The aim of the Odense training studies is to elucidate if regular physical training influences the expected decline in physical functional ability in order to assess capacity for postponing dependence in old age. All participants were healthy community-dwelling women representing three different age-cohorts of 65, 75 and 85 year-old subjects. The 65 and 85 year-old participants of the training group took part in physical class-based exercises for eight months with one session of 60 min a week, whereas the 75 year-old women trained twice a week over eight months. The multicomponent training consisted of various exercises typically involving body awareness, rhythm, aerobic performance (walking), muscle strength and muscle endurance, flexibility, reaction and balance exercises. Physical ability was measured as physical performance test (PPT), isometric muscle strength of the trunk, hip and leg, aerobic capacity and walking speed. This shows that regular training can significantly improve physical ability of elderly women with regard to PPT, maximal oxygen uptake and maximal walking speed, suggesting that both young-old and old-old women are able to benefit from regular tailored exercise training. Thus, physical training of old community dwelling women appears to represent a prophylactic remedy that merits further research aimed at evaluating the preliminary findings of the present studies in larger, less selective groups of participants. PMID- 12535321 TI - Exercise and injury prevention in older people. AB - This review aims to provide the reader with up to date evidence in relation to the role of exercise in the reduction of risk factors and the prevention of falls and injuries. Falls and injury may lead to a spiral of inactivity and decline that take older people close to or below the critical "thresholds" of performance necessary for everyday activities. Yet, low strength and power, poor balance, poor gait and functional ability, and fear of falling are all risk factors for falls modifiable with tailored exercise. Although the evidence on types, amounts and specificity of exercise to prevent falls is not complete, recommendations have been published that have been effective, either as an exercise stand-alone intervention or with exercise as part of a multifactorial intervention. It is clear that the target population must be at risk or already fallers, they must be "not too fit" and "not too frail". Supervised home-based exercise programs may be effective in those aged over 80 because they fall more frequently, injure more easily, and recover more slowly. In younger, community-dwelling, fallers multifactorial group interventions including targeting of balance, strength, power, gait, endurance, flexibility, co-ordination and reaction may be more effective. There are, however, research questions that still need answering - whether there are certain types of exercise harmful in certain subgroups of older people, what is the ideal intensity, frequency and duration of exercise for different subgroups of older people (primary and secondary prevention) and the relative value of the different components of fitness to prevention of falls and injuries. This review highlights the necessity of tailored, specific balance and strength exercise in the multidisciplinary prevention of falls and injuries. PMID- 12535322 TI - Unilateral and bilateral vasectomy in the dog: alkaline phosphatase as an indicator of tubular patency. AB - The objective of this research was to use a model of unilateral and bilateral occlusion of the ductus deferens in the dog to study the use of alkaline phosphatase (AP) as an indicator of tubular patency. Seven healthy cross bred dogs weighing 10-15 kg BW with normal spermiogram and AP concentrations in semen were used. From each dog, three semen samples were obtained before (intact) and after right (unilateral) and left (bilateral) vasectomy. The AP concentrations were measured in duplicates by a colorimetric method in each of the three fractions (first, second (sperm-rich), third) of each ejaculate. In addition, a macroscopic and microscopic evaluation of each ejaculate was carried out to assure its quality. Data were analysed by least squares analysis of variance using SAS(R). In intact and unilateral vasectomized dogs, 96.6% of AP measured in semen corresponded to the second sperm-rich fraction whereas 1.53 and 1.83% corresponded to the first and third fractions respectively. Total AP concentrations (first and second and third fraction) in vasectomized dogs were lower than in intact animals (19.857 vs 2284.431 +/- 4.347 UAL; p < 0.001). AP concentrations were much lower in bilateral than in unilateral vasectomized dogs (142 vs 39.572 +/- 4.347 UL, p < 0.001). In summary, AP concentrations in semen can be used as an early indicator of unilateral or bilateral lack of patency of the epididymal and deferent ducts in the dog. PMID- 12535323 TI - Immunohistochemical studies on oestrogen receptor alpha (ER alpha) and the proliferative marker Ki-67 in the sow uterus at different stages of the oestrous cycle. AB - In order to better understand physiological changes during the different stages of the oestrous cycle, immunohistochemistry was used in the present study to investigate the distribution of oestrogen receptor alpha (ER alpha) as well as the proliferative marker Ki-67, in the sow uterus during the oestrous cycle. Uterine samples were collected from multiparous sows with normal reproductive performance at selected stages of the oestrous cycle: at late dioestrus (d 17), prooestrus (d 19), oestrous (d 1), early dioestrus (d 4) and dioestrus (d 11-12), respectively. The tissue samples were fixed in 10% formaldehyde, embedded in paraffin and subjected to immunohistochemistry using monoclonal antibodies against ER alpha (C-311) and Ki-67 (MM-1). In general, the immunostaining of both ER alpha and Ki-67 was confined to nuclei of the target cells. Variations were seen, not only at the different stages of the oestrous cycle, but also in the different tissue compartments of the uterus. In the epithelia, the strongest ER alpha staining and highest amount of positive Ki-67 cells were found at early dioestrus. In the myometrium, the highest levels of staining of both ER alpha and Ki-67 positive cells were found at pro-oestrus and oestrus. For the proliferative marker, Ki-67, no positive cells were found at dioestrus and late dioestrus in the epithelium and myometrium. In the connective tissue stroma (subepithelial layer), the highest number of ER alpha positive cells were found at oestrus, which was significantly different compared with other stages (p< or = 0.05), whereas the levels of Ki-67 positive cells were relatively low and did not differ between the stages examined. Significant correlations between the number of ER alpha positive cells in the stroma and Ki-67 positive cells in the epithelia were observed. This suggests indirect regulatory mechanisms on epithelial proliferation via ER alpha in the stroma. In conclusion, these findings in the sow uterus show that the presence of ER alpha as well as Ki-67 protein varies not only between different stages of the oestrous cycle but also between different tissue compartments of the uterus. These findings indicate various regulatory mechanisms and stress the importance of localising ER alpha and proliferating cells in different uterine tissues. PMID- 12535324 TI - Investigations on the re-establishment of the positive feedback of oestradiol during anoestrus in the bitch. AB - To test for the re-establishment of the positive feedback of oestradiol (E2) during anoestrus in the dog, the hypothalamo-pituitary-ovarian axis of five beagle bitches was challenged by treatments with oestradiol benzoate (EB), mimicking the course of the pro-oestric E2 secretion. Treatments in anoestrus started 7 days following the decline of progesterone (P) <1 ng/ml; they were repeated in 5 week intervals until onset of pro-oestrus; another treatment was performed during dioestrus 50 days after onset of the preceding pro-oestric bleeding. Each dog served as its own control by receiving vehicle-treatments in one of the following cycles. Each observation period covered a time window of 168 h and blood samples were collected for the determination of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and E2 in 6 (0-24 h) and 8 h (24-168 h) intervals. In the control periods and as indicated by the parameters area under curve (AUC), basal and maximal values, the availability of LH, FSH and E2 decreased from dioestrus to early anoestrus to increase again during the course of anoestrus (p < 0.05), indicating a gradual desensitization of the hypothalamus towards the negative feedback of oestradiol. At all times treatments with EB lowered the availability of FSH (decreased AUC and basal levels). A delay in the occurrence of the first LH peak after treatments with EB (p < 0.001) and decreased maximal values (p < 0.001) indicated a suppression of the LH-release. In no case treatment with EB led to a pre-ovulatory like LH-surge. In each dog the last trial with EB in anoestrus passed over into pro-oestrus/oestrus, with a reduced AUC and peak value of the pre-ovulatory LH-surge being the only differences to the control group. The observed differences in the response of LH and FSH to treatments with EB point towards subtle differences in the mechanisms controlling the release of these two hormones during anoestrus. From the data obtained, it may be concluded that the time window for E2 to act via a positive feedback seems to be very small and restricted to the end of anoestrus, and that full follicular function is a pre-requisite to allow for this phenomenon. PMID- 12535325 TI - In vitro maturation and fertilization of buffalo oocytes: effects of storage of ovaries, IVM temperatures, storage of processed sperm and fertilization media. AB - Studies were conducted to examine the possibility of preserving slaughterhouse derived buffalo ovaries at 4 degrees C for 0 (control), 12 and 24 h to maintain the developmental competence of the oocytes (experiment 1), to assess the effect of incubation temperature during oocyte maturation on rates of in vitro maturation (IVM) and in vitro fertilization (IVF) of buffalo oocytes and embryo development (experiment 2), and to examine the effect of storage at 25 degrees C for 0 (control), 4 and 8 h of frozen-thawed buffalo sperm and BO and H-TALP as sperm processing and fertilization media on cleavage and embryo development in vitro of buffalo oocytes (experiment 3) in order to optimize the IVF technology in buffalo. Results suggested that storage of ovaries at 4 degrees C for 12 or 24 h significantly (p < 0.05) reduced the developmental potential of oocytes. Incubation temperatures during the IVM influenced the fertilization rate but had no significant effect on maturation and subsequent embryo development. The incubation temperature of 38.5 degrees C during IVM was found to be optimum for embryo production in vitro. Storage of frozen-thawed sperm at 25 degrees C for 8 h significantly (p < 0.05) decreased its ability to cleave the oocytes. Sperm processed in BO medium had significantly (p < 0.05) higher ability to cleave the oocytes than the H-TALP medium. PMID- 12535326 TI - Studies on the intracellular Ca2+ concentration of thawed dog spermatozoa: influence of Equex from different sources, two thawing diluents and post-thaw incubation in capacitating conditions. AB - The addition of 0.5% (v/v) of Equex STM Paste (Nova Chemical Sales, Scituate Inc., MA, USA), whose active ingredient is sodium dodecyl sulphate (SDS), to a Tris-egg yolk extender was demonstrated to improve the longevity of frozen-thawed dog spermatozoa during in vitro incubation at 38 degrees C. The aim of the first experiment was to compare the effects of two SDS-containing compounds, Equex STM Paste and Equex Pasta (Minitub, Tiefenbach, Germany), when added to a Tris-egg yolk based extender, on the post-thaw longevity of dog spermatozoa, as well as on the intracellular Ca2+ concentration of spermatozoa, during post-thaw incubation at 38 degrees C. The post-thaw sperm survival and longevity, as well as the quality of the sperm movement, were significantly better when using Equex STM Paste. Such prolonged sperm longevity, however, was associated to a higher intracellular Ca2+ concentration in a large subpopulation of the live spermatozoa. A second experiment was aimed to evaluate the effects of sperm dilution immediately post-thaw with a Tris buffer containing glucose or fructose. The two Tris buffers were no different for any of the sperm parameters studied. The aim of a third experiment was to evaluate the sperm longevity, motility patterns and intracellular Ca2+ concentration of cryopreserved dog spermatozoa during post-thaw incubation in capacitating conditions [canine capacitating medium (CCM) with or without 5 microg/ml of heparin]. Heparin had no significant effects on any of the sperm parameters evaluated. During the first 8 h of incubation, the majority of the live spermatozoa had a high intracellular Ca2+ content. However, after 8-10 h of incubation, it had significantly declined. The highest proportion of fast motile sperm, and the highest curvilinear velocity, average path velocity and amplitude of lateral head displacement for the total motile sperm were observed during the 2-4-h incubation period. It was concluded that: (a) the addition of 0.5% (v/v) of Equex STM Paste to a Tris-egg yolk based extender significantly improved the post-thaw longevity of dog spermatozoa, but the same concentration of Equex Pasta had no significant beneficial effects; (b) sperm dilution after thawing with a Tris buffer containing glucose or fructose made no difference in post-thaw sperm longevity; (c) the addition of 5 microg/ml of heparin to CCM had no significant capacitating effects on frozen-thawed dog spermatozoa. PMID- 12535327 TI - Improving the reproductive efficiency, pregnancy diagnosis and monitoring the resumption of luteal activity in indigenous damascus goats. AB - Seventy-five female Damascus goats aged between 1.5 and 5.5 years were used to evaluate the effectiveness of the intravaginal sponges and prostaglandin analogue on oestrous synchronization and fecundity; to diagnose pregnancy and to monitor the resumption of the luteal activity. Females were divided randomly, during the breeding season, into three equal groups, S, P and C. Females in group S were fitted with sponges containing 45 mg of flugestone acetate (FGA) for 14 days and injected with pregnant mare serum gonadotrophin (PMSG) at the sponge withdrawal. Females in group P were given two injections of prostaglandin F2alpha analogue at 11-day intervals, whereas females in group C (control) received no treatment. The results showed that there was a significant difference (p < 0.001) in oestrous exhibition between females in group S as compared with those in groups P and C, with means being 30 +/- 10, 172 +/- 115 and 217 +/- 75 h for groups S, P and C, respectively. Kidding rates resulting from the first and all matings were 80 and 88, 52 and 88, and 68 and 80% for groups S, P and C, respectively. Fecundity rates were 215, 175 and 180% for groups S, P and C, respectively, with a significant difference (p < 0.05) between the S and both P and C groups. Using an ultrasound pregnancy detector performed on days 57 +/- 3 after mating, positive pregnancy diagnosis was 93.3% and 100% for non-pregnancy. Females in the control group showed functional corpus luteum starting in September. It is concluded that FGA sponges plus PMSG treatment could be successfully used to synchronize oestrus and improve fecundity; whereas prostaglandin treatment was not effective to synchronize oestrus. It is also concluded that pregnancy can be diagnosed accurately and successfully using an ultrasound pregnancy detector. In addition, ovarian activity in the Damascus goat in Syria resumes in September. PMID- 12535328 TI - Time empty and ketone body status in the early postpartum period of dairy cows. AB - This study aimed to assess the effect of ketone body status early postpartum on time empty in 74 multiparous dairy cows under field conditions. Animals were equally distributed across eight farms and were controlled by the same herd fertility-monitoring programme. Cows were visited twice antepartum and six times postpartum at weekly intervals between 0530 and 0830 a.m. On these occasions, body condition scores and milk yields were measured, blood and milk samples were taken, cows were gynaecologically examined, and parameters of reproduction were recorded. Cows with a time empty of less or more than 80 days were classified as early and late conceiving cows (EC and LC, respectively). A time empty of 80 days results in calving-to-calving intervals of 1 year and classification based on this threshold value resulted in groups of equal size and equal distribution of EC and LC within farms. Ketone bodies measured were beta-hydroxybutyrate in blood and acetoacetate and acetone in blood and milk. Blood and milk ketone body concentrations, as well as the ratios of acetoacetate and acetone to beta hydroxybutyrate, over the first 6 weeks postpartum were higher in LC than in EC, whereas plasma glucose and non-esterified fatty acids and milk fat, protein and urea concentrations did not exhibit clear differences between groups. Ketone body concentrations were as good predictors of time empty as ketone body ratios and might have practical impact in herd-fertility-monitoring programmes. PMID- 12535329 TI - Oncology and ethics. PMID- 12535330 TI - Effects of ram introduction after the second prostaglandin F2 alpha injection on day 11 on the LH surge characteristics in fat-tailed ewes. AB - The aim of this study was to evaluate the effects of ram introduction after the second prostaglandin F2alpha (PG F2alpha) injection on day 11 on the secretion characteristics of pre-ovulatory LH surge of fat-tailed ewes. Multiparous Morkaraman ewes (n=12) were divided into three groups by balancing the groups for liveweight (BW) and body condition score (BCS). On the day of second PGF2 alpha injection (0 h), performance tested rams (n=2) were either introduced to the ewes at 0 h (ram 0 group, n=4) or at 18 h (ram 18 group, n=4) or were not introduced (control group, n=4). Blood samples were collected at 6, 18, 42, 48, 56, 62, 66, 70, 74, 78 and 90 h for the determination of pre-ovulatory LH surge. BCS and BW during the experimental period were 2.2 +/- 0.2 units and 50.9 +/- 2.3 kg, 2.4 +/ 0.4 units and 49.2 +/- 6.2 kg, 2.1 +/- 0.3 units and 45.9 +/- 4.4 kg, respectively for the ram 0, ram 18 and control groups (p > 0.05). No significant difference was observed in LH surge characteristics for the experimental groups. Peak LH concentrations were also not different between groups (p > 0.05) and they were 12.2 +/- 8.3, 29.1 +/- 9.9 and 15.8 +/- 9.5 microg/l for the ram 0, ram 18 and control groups, respectively. There was, however, a significant correlation between peak LH concentrations and BCS (p < 0.05, R2=0.373). In conclusion, it appears that, compared with ram introduction, variability in body condition of the ewe has much pronounced effect on the amount of LH secreted after the usage of two PGF2 alpha injections (11 days apart) as a tool for oestrus synchronization. PMID- 12535331 TI - Meiotic response of in vitro matured canine oocytes under different proteins and heterologous hormone supplementation. AB - The impact of TCM-199 supplemented with different proteins and heterologous hormones on the in vitro maturation (IVM) rate of bitch oocytes was evaluated by nuclear staining under fluorescence microscopy. Oocytes were recovered by slicing of ovaries from bitches presented at various stages of oestrous cycle to ovariohysterectomy. The basic culture medium was TCM-199 supplemented with 25 mM Hepes/l, with 10% heat-inactivated oestrous cow serum (ECS), 50 microg/ml gentamicin, 2.2 mg/ml sodium bicarbonate and 22-microg/ml pyruvic acid, 1.0 microg/ml oestradiol (E 8875; Sigma), 0.5-microg/ml follicle-stimulating hormone (FSH) (Folltropin-V; Vetrepharm Inc., Ontario, Canada) and 0.03 IU/ml human gonadotropin (hCG) (Profasi HP; Serono, Aubonne, Switzerland). Oocytes were distributed randomly between basic culture medium (control) and the corresponding experimental treatment. Hormone treatments were: oocytes cultured in; (1) medium without FSH, (2) control medium supplemented with 20 microg/ml oestradiol, or (3) medium supplemented with 1 microg/ml human somatotropin (hST; Humatrope, Lilly, Saint Cloud, France). The second experiment consisted of oocytes cultured in medium supplemented with 0.4% (w/v) bovine serum albumin (BSA, fraction V; Gibco Grand Island, NY, USA) instead of ECS, or oocytes cultured in medium with 10% inactivated oestrous bitch serum (EBS) instead of ECS. Oocytes were cultured in 100 microl droplets (up to 25 oocytes per drop) under mineral oil at 37 degrees C in a 100% humidified atmosphere containing 5% CO2 in air. After 72 h of IVM, the highest rates (p < 0.05) of meiotic resumption were achieved with the 0.4% BSA supplementation. A positive influence on the metaphase II (MII) acquisition rate was observed with hST supplement. Oocytes cultured with 10% EBS supplementation did not develop to the MII stage. The results in this study show that the protein and hormone supplements to TCM-199 culture medium tested did not promote the final steps of IVM of bitch oocytes. PMID- 12535332 TI - Sialomucin complex (Muc4) expression in porcine endometrium during the oestrous cycle and early pregnancy. AB - The non-invasive type of implantation in the pig is characterized by the maintenance of a thick glycocalyx coating on the uterine epithelial surface microvilli. Present study investigated the alteration in the sialomucin complex (Muc4) expression during the oestrous cycle and early pregnancy in the pig. Endometrial tissue samples were immunostained with the primary antibody to the Muc4 transmembrane subunit ASGP-2. Muc4 immunostaining increased in the surface and glandular epithelia between days 5 and 10 of oestrous cycle. Immunostaining continued to increase on day 12 with the greatest intensity of uterine Muc4 immunostaining detected on day 15 of the oestrous cycle and early pregnancy. Endometrial Muc4 expression in cyclic gilts decreased dramatically during early proestrous but continued to remain abundant in the surface and glandular epithelium of pregnant gilts during the period of conceptus attachment to the uterine surface. PMID- 12535333 TI - Analysis of in vitro fertilizing capacity to evaluate the freezing procedures of boar semen and to predict the subsequent fertility. AB - A porcine in vitro fertilization (IVF) system and seminal quality parameters of frozen-thawed boar semen were used to assess the effectiveness of two different thawing rates of frozen boar semen, and to address the question of whether differences between fertility of ejaculates could be predicted in a limited field trial. In the first experiment, two thawing procedures were analysed (37 degrees C, 30 s; 50 degrees C, 12 s) and no differences in sperm quality were found. However, when the procedure was 50 degrees C, 12 s the IVF results showed a higher number of sperm per penetrated oocyte and a near 10 points higher rate of pronuclear formation. In the second experiment, the fertility results obtained in the limited field trial show to be efficient enough for application in a commercial use, especially for three of the employed boars (fertility > or = 80%). In this limited study, the conventional seminal parameters are not accurate enough to discriminate good and bad boars in relation to fertility. On the contrary, parameters of in vitro penetrability are more precise to predict subsequent fertilities. As conclusion, the IVF fertilization system seems to be a good tool to evaluate the quality of frozen-thawed boar semen previous to its commercial way, to verify the bank semen storage quality and a good way to assay new sperm freezing procedures, as it is the more precise evaluating method in estimating the potential fertilizing ability. PMID- 12535334 TI - Ethics and genetic selection in purebred dogs. AB - There is an ongoing revolution in medicine that is changing the way that veterinarians will be counselling clients regarding inherited disorders. Clinical applications will emerge rapidly in veterinary medicine as we obtain new information from canine and comparative genome projects (Meyers-Wallen 2001: Relevance of the canine genome project to veterinary medical practice. International Veterinary Information Service, New York). The canine genome project is described by three events: mapping markers on canine chromosomes, mapping gene locations on canine chromosomes (Breen et al. 2001: Genome Res. 11, 1784-1795), and obtaining the nucleotide sequence of the entire canine genome. Information from such research has provided a few DNA tests for single gene mutations [Aguirre 2000: DNA testing for inherited canine diseases. In: Bonagura, J (ed), Current Veterinary Therapy XIII. Philadelphia WB Saunders Co, 909-913]. Eventually it will lead to testing of thousands of genes at a time and production of DNA profiles on individual animals. The DNA profile of each dog could be screened for all known genetic disease and will be useful in counselling breeders. As part of the pre-breeding examination, DNA profiles of prospective parents could be compared, and the probability of offspring being affected with genetic disorders or inheriting desirable traits could be calculated. Once we can examine thousands of genes of individuals easily, we have powerful tools to reduce the frequency of, or eliminate, deleterious genes from a population. When we understand polygenic inheritance, we can potentially eliminate whole groups of deleterious genes from populations. The effect of such selection on a widespread basis within a breed could rapidly improve health within a few generations. However, until we have enough information on gene interaction, we will not know whether some of these genes have other functions that we wish to retain. And, other population effects should not be ignored. At least initially it may be best to use this new genetic information to avoid mating combinations that we know will produce affected animals, rather than to eliminate whole groups of genes from a population. This is particularly important for breeds with small gene pools, where it is difficult to maintain genetic diversity. Finally, we will eventually have enough information about canine gene function to select for specific genes encoding desirable traits and increase their frequencies in a population. This is similar to breeding practices that have been applied to animals for hundreds of years. The difference is that we will have a large pool of objective data that we can use rapidly on many individuals at a time. This has great potential to improve the health of the dog population as a whole. However, if we or our breeder clients make an error, we can inadvertently cause harm through massive, rapid selection. Therefore, we should probably not be advising clients on polygenic traits or recommend large scale changes in gene frequencies in populations until much more knowledge of gene interaction is obtained. By then it is likely that computer modelling will be available to predict the effect of changing one or several gene frequencies in a dog population over time. And as new mutations are likely to arise in the future, these tools will be needed indefinitely to detect, treat and eliminate genetic disorders from dog populations. Information available from genetic research will only be useful in improving canine health if veterinarians have the knowledge and skills to use it ethically and responsibly. There is not only a great potential to improve overall canine health through genetic selection, but also the potential to do harm if we fail to maintain genetic diversity. Our profession must be in a position to correctly advise clients on the application of this information to individual dogs as well as to populations of dogs, and particularly purebred dogs. PMID- 12535336 TI - Reduced amino acid content in transgenic potato tubers due to antisense inhibition of the leaf H+/amino acid symporter StAAP1. AB - Transport processes across the plasma membrane of leaf vascular tissue are essential for transport and distribution of assimilates. In potato, leaves are the predominant sites for nitrate reduction and amino acid biosynthesis. From there, assimilated amino acids are exported through the phloem to supply tubers with organic nitrogen. To study the role of amino acid transporters in long distance transport and allocation of organic nitrogen in potato plants, a gene encoding a functional, leaf-expressed amino acid permease StAAP1 was isolated. Similar to the sucrose transporter SUT1, StAAP1 expression was induced during the sink-to-source transition, indicating a role in phloem loading. To test the role of StAAP1, expression was inhibited by an antisense approach. Transgenic plants with reduced StAAP1 expression were phenotypically indistinguishable from wild type, as were photosynthetic capacity and tuber yield. However, tubers from antisense StAAP1 plants showed up to 50% reduction in free amino acid contents. In comparison, starch content was not affected or tended to increase relative to wild type. The reduction in all amino acids except aspartate in the antisense plants is consistent with the properties of amino acid permeases (AAPs) found in heterologous systems. The results demonstrate an important role for StAAP1 in long-distance transport of amino acids and highlight the importance of plasma membrane transport for nutrient distribution in plants. PMID- 12535337 TI - Biochemical and functional analysis of CTR1, a protein kinase that negatively regulates ethylene signaling in Arabidopsis. AB - CTR1 encodes a negative regulator of the ethylene response pathway in Arabidopsis thaliana. The C-terminal domain of CTR1 is similar to the Raf family of protein kinases, but its first two-thirds encodes a novel protein domain. We used a variety of approaches to investigate the function of these two CTR1 domains. Recombinant CTR1 protein was purified from a baculoviral expression system, and shown to possess intrinsic Ser/Thr protein kinase activity with enzymatic properties similar to Raf-1. Deletion of the N-terminal domain did not elevate the kinase activity of CTR1, indicating that, at least in vitro, this domain does not autoinhibit kinase function. Molecular analysis of loss-of-function ctr1 alleles indicated that several mutations disrupt the kinase catalytic domain, and in vitro studies confirmed that at least one of these eliminates kinase activity, which indicates that kinase activity is required for CTR1 function. One missense mutation, ctr1-8, was found to result from an amino acid substitution within a new conserved motif within the N-terminal domain. Ctr1-8 has no detectable effect on the kinase activity of CTR1 in vitro, but rather disrupts the interaction with the ethylene receptor ETR1. This mutation also disrupts the dominant negative effect that results from overexpression of the CTR1 amino-terminal domain in transgenic Arabidopsis. These results suggest that CTR1 interacts with ETR1 in vivo, and that this association is required to turn off the ethylene-signaling pathway. PMID- 12535338 TI - The GLOBULAR ARREST1 gene, which is involved in the biosynthesis of folates, is essential for embryogenesis in Arabidopsis thaliana. AB - We identified a mutation in Arabidopsis that resulted in defective embryos, and we designated this mutation globular arrest1 (gla1). The predicted amino acid sequence encoded by the GLA1 gene is homologous to the amino acid sequences of folylpolyglutamate synthetase (FPGS) and dihydrofolate synthetase (DHFS), which participate in folate biosynthesis. The defect of gla1 in the formation of calli was rescued by the supplement of 5-formyl tetrahydrofolate. These results indicated that GLA1 is involved in the biosynthesis of tetrahydrofolate. The gla1 embryos developed normally in the early stage of development but did not undergo the transition to the heart stage. Thus, the function of the GLA1 gene in embryogenesis appears to be required after the globular stage. However, when the levels of GLA1 transcripts in transgenic plants were increased by introduction of several copies of a GLA1 transgene (GLA6.8), the gla1 embryos that grew on gla1/gla1 GLA6.8/- plants developed as far as the heart to bent-cotyledon stage. This result suggests that the GLA1 function is provided to embryos by maternal tissues until embryos reach the globular stage. PMID- 12535339 TI - Susceptible to intolerance--a range of hormonal actions in a susceptible Arabidopsis pathogen response. AB - Ethylene and salicylic acid (SA) are key intermediates in a host's response to pathogens. Previously, we have shown using a tomato compatible interaction that ethylene and SA act sequentially and are essential for disease symptom production. Here, we have examined the relationship between the two signals in the Arabidopsis-Xanthomonas campestris pv. campestris (Xcc) compatible interaction. Preventing SA accumulation by expression of the nahG gene reduced subsequent ethylene production and altered the development of disease symptoms, with plants showing no visible chlorosis. The ethylene insensitive lines, etr1-1 and etr2-1, on the other hand, accumulated SA and exhibited normal but precocious symptom development. Therefore, Arabidopsis, like tomato, was found to exhibit co operative ethylene and SA action for the production of disease symptoms. However, in Arabidopsis, SA was found to act upstream of ethylene. Jasmonic acid and indole-3-acetic acid levels were also found to increase in response to Xcc. In contrast to ethylene, accumulation of these hormones was not found to be dependent on SA action. These results indicate that the plants response to a virulent pathogen is a composite of multiple signaling pathways. PMID- 12535340 TI - Two different novel cis-acting elements of erd1, a clpA homologous Arabidopsis gene function in induction by dehydration stress and dark-induced senescence. AB - Many plant genes have been shown to be induced by water stress and function in stress tolerance. The erd1 gene has been shown to be upregulated in response to both water stress and etiolation. Promoter studies using the erd1 promoter region fused to the luciferase (LUC) reporter gene in Arabidopsis thaliana were performed to identify the putative cis elements involved. Results indicated that the cis elements, responsible for gene expression during dehydration and etiolation, are separately located in two discrete portions of the erd1 promoter. Base substitution analysis showed that a 14-bp region from -599 to -586, and a myc recognition motif from -466 to -461 are necessary for the induction of LUC activity in dehydrated plants. On the other hand, base substitution analysis revealed that both an abscisic acid responsive element (ABRE)-like sequence (from -199 to -195) and an ACGT sequence (from -155 to -152) are required for an etiolation-induced increase in LUC activity. LUC activity measurements from etiolated transgenic plants incubated in either water, N6-benzyleadenine (BA), or a 1% sucrose solution found that while BA was able to delay the increase in LUC activity seen in water-treated plants, no increase in LUC activity was seen in plants incubated in sucrose. These results indicate that the erd1 promoter contains two different regulatory systems that are involved in upregulation by dehydration stress and dark-induced senescence. PMID- 12535341 TI - Diverse RNA viruses elicit the expression of common sets of genes in susceptible Arabidopsis thaliana plants. AB - Systemic infections of plants by viruses require that viruses modify host cells in order to facilitate infections. These modifications include induction of host factors required for replication, propagation and movement, and suppression of host defense responses, which are likely to be associated with changes in host gene expression. Past studies of the effects of viral infection on gene expression in susceptible hosts have been limited to only a handful of genes. To gain broader insight into the responses elicited by viruses in susceptible hosts, high-density oligonucleotide probe microarray technology was used. Arabidopsis leaves were either mock inoculated or inoculated with cucumber mosaic cucumovirus, oil seed rape tobamovirus, turnip vein clearing tobamovirus, potato virus X potexvirus, or turnip mosaic potyvirus. Inoculated leaves were collected at 1, 2, 4, and 5 days after inoculation, total RNA was isolated, and samples were hybridized to Arabidopsis GeneChip microarrays (Affymetrix). Microarray hybridization revealed co-ordinated changes in gene expression in response to infection by diverse viruses. These changes include virus-general and virus specific alterations in the expression of genes associated with distinct defense or stress responses. Analyses of the promoters of these genes further suggest that diverse RNA viruses elicit common responses in susceptible plant hosts through signaling pathways that have not been previously characterized. PMID- 12535342 TI - Transgenic expression of a putative calcium transporter affects the time of Arabidopsis flowering. AB - PPF1 is a gibberellin-induced, vegetative growth-specific gene, first isolated from short-day (SD)-grown G2 pea plants. In the current work, we found that transgenic Arabidopsis plants overexpressing the PPF1 gene (PPF1 (+)) flowered much later and had a significantly longer lifespan compared to control plants, whereas suppression of this gene (PPF1 (-)) resulted in a very rapid reproductive cycle. Western blotting analyses of PPF1 (+) and (-) plant lines revealed a positive correlation between the amount of antibody-reactive protein and the time of flowering. Green fluorescent protein (GFP) co-expression assays showed that the PPF1 protein is likely localized in chloroplast membranes. Transgenic expression of PPF1 affected the calcium storage capacities since chloroplasts isolated from PPF1 (+) plants contained high Ca2+ levels while chloroplasts of PPF1 (-) plants contained very low amounts of calcium ion. Using Novikoff human hepatoma cells, we demonstrated that expression of PPF1 leads to a significant inward calcium ion current that was absent in untransformed cells. We conclude that, as a putative calcium ion carrier, PPF1 affects the flowering time of higher plants by modulating Ca2+ storage capacity within chloroplasts. PMID- 12535343 TI - Leaves of the Lamiaceae species Glechoma hederacea (ground ivy) contain a lectin that is structurally and evolutionary related to the legume lectins. AB - A novel lectin has been isolated and cloned from leaves of Glechoma hederacea (ground ivy), a typical representative of the plant family Lamiaceae. Biochemical analyses indicated that the G. hederacea agglutinin (Gleheda) is a tetrameric protein consisting of four subunits pairwise linked through an interchain disulphide bridge and exhibits a preferential specificity towards N acetylgalactosamine. Cloning of the corresponding gene and molecular modeling of the deduced sequence demonstrated that Gleheda shares high sequence similarity with the legume lectins and exhibits the same overall fold and three-dimensional structure as the classical legume lectins. The identification of a soluble and active legume lectin ortholog in G. hederacea not only indicates that the yet unclassified Lamiaceae lectins belong to the same lectin family as the legume lectins, but also sheds a new light on the specificity, physiological role and evolution of the classical legume lectins. PMID- 12535344 TI - GeBP, the first member of a new gene family in Arabidopsis, encodes a nuclear protein with DNA-binding activity and is regulated by KNAT1. AB - Trichomes of Arabidopsis are single-celled epidermal hair that are a useful model for studying plant cell fate determination. Trichome initiation requires the activity of the GLABROUS1 (GL1) gene whose expression in epidermal and trichome cells is dependent on the presence of a 3'-cis-regulatory element. Using a one hybrid screen, we have isolated a cDNA, which encodes for a protein, GL1 enhancer binding protein (GeBP), that binds this regulatory element in yeast and in vitro. GeBP and its three homologues in Arabidopsis share two regions: a central region with no known motifs and a C-terminal region with a putative leucine-zipper motif. We show that both regions are necessary for trans-activation in yeast. A translational fusion with the Yellow Fluorescent Protein (YFP) indicates that GeBP is a nuclear protein whose localization is restricted to, on average, 3-5 subnuclear foci that might correspond to nucleoli. Transcriptional fusion with the GUS reporter indicates that GeBP is mainly expressed in vegetative meristematic tissues and in very young leaf primordia. We looked at GeBP expression in plants mutated in or misexpressing KNAT1, a KNOX gene, expressed in the shoot apical meristem and downregulated in leaf founder cells, and found that GeBP transcript level is regulated by KNAT1 suggesting that KNAT1 is a transcriptional activator of GeBP. This regulation suggests that GeBP is acting as a repressor of leaf cell fate. PMID- 12535345 TI - Arabidopsis actin gene ACT7 plays an essential role in germination and root growth. AB - Arabidopsis contains eight actin genes. Of these ACT7 is the most strongly expressed in young plant tissues and shows the greatest response to physiological cues. Adult plants homozygous for the act7 mutant alleles show no obvious above ground phenotypes, which suggests a high degree of functional redundancy among plant actins. However, act7-1 mutant plants are at a strong selective disadvantage when grown in competition with wild-type plants and therefore must have undetected physical defects. The act7-1 and act7-4 alleles contain T-DNA insertions just after the stop codon and within the first intron, respectively. Homozygous mutant seedlings of both alleles showed less than 7% of normal ACT7 protein levels. Mutants displayed delayed and less efficient germination, increased root twisting and waving, and retarded root growth. The act7-4 mutant showed the most dramatic reduction in root growth. The act7-4 root apical cells were not in straight files and contained oblique junctions between cells suggesting a possible role for ACT7 in determining cell polarity. Wild-type root growth was fully restored to the act7-1 mutant by the addition of an exogenous copy of the ACT7 gene. T-DNA insertions just downstream of the major polyadenylation sites (act7-2, act7-3) appeared fully wild type. The act7 mutant phenotypes demonstrate a significant requirement for functional ACT7 protein during root development and explain the strong negative selection component seen for the act7-1 mutant. PMID- 12535346 TI - SAD: a new DOF protein from barley that activates transcription of a cathepsin B like thiol protease gene in the aleurone of germinating seeds. AB - Functional analysis of hydrolase gene promoters, induced by gibberellin (GA) in aleurone cells following germination, has identified a GA-responsive complex (GARC) as a tripartite element containing a pyrimidine-box motif 5'-CCTTTT-3'. We describe here the characterization of a new barley gene (Sad gene) encoding a transcription factor (SAD) of the DNA binding with One Finger (DOF) class that binds to the pyrimidine box in vitro and activates transcription of a GA-induced protease promoter in bombarded aleurone layers. RT-PCR and in situ hybridization analyses showed that the Sad transcripts accumulated in all tissues analysed, being especially abundant in the scutellum and aleurone cells upon seed germination. The SAD protein, expressed in bacteria, binds in a specific manner to two oligonucleotides containing the sequence 5'-G/CCTTTT/C-3', derived from the promoter region of the Al21 gene encoding a cathepsin B-like cysteine protease. Although the Sad transcript accumulation did not respond to external GA incubation in aleurone cells, in transient expression experiments in co-bombarded aleurone layers, SAD trans-activated transcription from the Al21 promoter in a similar manner as did GAMYB, a MYB protein previously shown to respond to GA and to activate several hydrolase gene promoters in barley aleurone cells. In vivo interaction between the GAMYB and SAD proteins was shown in the yeast two-hybrid system, where GAMYB potentiates the SAD trans-activation capacity through interaction with its C-terminal domain. PMID- 12535347 TI - De-submergence-induced ethylene production in Rumex palustris: regulation and ecophysiological significance. AB - Rumex palustris responds to total submergence by increasing the elongation rate of young petioles. This favours survival by shortening the duration of submergence. Underwater elongation is stimulated by ethylene entrapped within the plant by surrounding water. However, abnormally fast extension rates were found to be maintained even when leaf tips emerged above the floodwater. This fast post submergence growth was linked to a promotion of ethylene production that is presumed to compensate for losses brought about by ventilation. Three sources of ACC contributed to post-submergence ethylene production in R. palustris: (i) ACC that had accumulated in the roots during submergence and was transported in xylem sap to the shoot when stomata re-opened and transpiration resumed, (ii) ACC that had accumulated in the shoot during the preceding period of submergence and (iii) ACC produced de novo in the shoot following de-submergence. This new production of ethylene was associated with increased expression of an ACC synthase gene (RP ACS1) and an ACC oxidase gene (RP-ACO1), increased ACC synthase activity and a doubling of ACC oxidase activity, measured in vitro. Out of seven species of Rumex examined, a de-submergence upsurge in ethylene production was seen only in shoots of those that had the ability to elongate fast when submerged. PMID- 12535348 TI - Molecular properties of the putative nitrogen sensor PII from Arabidopsis thaliana. AB - Although the signal sensing protein PII is well known to play a central role in bacterial nitrogen metabolism, the structure and function of PII in plants remains only partially understood. Comparative modeling was undertaken based on the high degree of amino acid identity between Escherichia coli and Arabidopsis PII. The mature Arabidopsis PII predicted structure superimposes very well onto the E. coli PII structure (Calpha root mean square deviation < 0.4 A). The model of the highly conserved T-loop suggests a molecular mechanism by which the plant PII may regulate putative post-translational modification in response to metabolite binding. Consistent with the presence of key conserved residues necessary for trimer formation, gel filtration showed the oligomeric structure of Arabidopsis thaliana PII to be a homotrimer. We have demonstrated that Arabidopsis PII binds to the small molecules, ATP, ADP, 2KG, and with lesser affinity to OAA, using isothermal titration calorimetry. We have determined the metabolite dissociation constants and compared these with known physiological concentrations of these metabolites in the plant to identify the Arabidopsis PII effector molecules and their possible roles. We predict that the plant PII is likely continually bound by ATP, and its ligand-bound state only varying with respect to the degree of 2KG binding. Based on our in vitro binding studies, the function of plant PII as a 2KG sensor is suggested. PMID- 12535349 TI - The circadian clock regulated RNA-binding protein AtGRP7 autoregulates its expression by influencing alternative splicing of its own pre-mRNA. AB - The clock-regulated RNA-binding protein AtGRP7 is part of a negative feedback circuit through which the protein influences circadian oscillations of its own transcript. Constitutive overexpression of AtGRP7 in transgenic plants leads to the appearance of a low amount of an alternatively spliced Atgrp7 transcript with a premature stop codon. It is generated by the use of a 5' cryptic splice site in the middle of the intron at the expense of the fully spliced mRNA, indicating a role for AtGRP7 in splice site selection. Accelerated decay of this transcript accounts for its low steady state abundance. This implicates a mechanism for the AtGRP7 feedback loop: Atgrp7 expression is downregulated, as AtGRP7 protein accumulates over the circadian cycle, partly by the generation of an alternate transcript that due to its instability does not accumulate to high levels and does not produce a functional protein. Recombinant AtGRP7 protein specifically interacts with the 3' untranslated region and the intron of its transcript, suggesting that the shift in splice site selection and downregulation involves binding of AtGRP7 to its pre-mRNA. AtGRP7 also influences the choice of splice sites in the Atgrp8 transcript encoding a related RNA-binding protein, favoring the production of an alternatively spliced, unstable Atgrp8 transcript. This conservation points to the importance of this regulatory mechanism to control the level of the clock-regulated glycine-rich RNA-binding proteins and shows how AtGRP7 can control abundance of target transcripts. PMID- 12535350 TI - The DNA-binding activity of an AP2 transcriptional activator HvCBF2 involved in regulation of low-temperature responsive genes in barley is modulated by temperature. AB - An HvCBF2 cDNA was isolated from barley leaves. It encoded a protein containing an AP2 DNA-binding domain homologous to C-repeat (CRT)/dehydration-responsive element (DRE) binding factors (CBF/DREB1). In contrast to the previously reported cold-inducible CBF/DREB1 genes, HvCBF2 was expressed in barley leaves under non stress conditions. Only a transient increase in the HvCBF2 transcript level was observed during cold treatment. Transactivation analysis showed that HvCBF2 was a transcriptional activator, capable of activating expression of a reporter gene driven by a low-temperature and drought-responsive HVA1s promoter in barley leaves. The activity of HvCBF2 as a transcriptional activator was upregulated by low temperature. DNA-binding analysis revealed that HvCBF2 did not bind to the CRT/DRE motif at 30 degrees C. A low, but detectable, binding activity was observed at 25 degrees C and the binding activity gradually increased as the temperature decreased. The binding activity at 0 degrees C was the highest and more than 10 times higher than that at 25 degrees C. The activation and inactivation of HvCBF2 activity were reversible and were achieved in a cell-free system simply by temperature change. Analysis of the binding sequence showed that HvCBF2 bound to a (G/a)(T/c)CGAC core motif, where the lower-case letters are less efficient bases. These data suggest that HvCBF2 is a transcription factor interacting with the core CRT/DRE motif containing a preferred sequence of GTCGAC and its DNA-binding activity is regulated by temperature. This represents a new type of activation mechanism for transcriptional activators. PMID- 12535351 TI - Serine/threonine kinase activity in the putative histidine kinase-like ethylene receptor NTHK1 from tobacco. AB - A histidine kinase-based signaling system has been proposed to function in ethylene signal transduction pathway of plants and one ethylene receptor has been found to possess His kinase activity. Here we demonstrate that a His kinase-like ethylene receptor homologue NTHK1 from tobacco has serine/threonine (Ser/Thr) kinase activity, but no His kinase activity. Evidence obtained by analyzing acid/base stability, phosphoamino acid and substrate specificity of the phosphorylated kinase domain, supports this conclusion. In addition, mutation of the presumptive phosphorylation site His (H378) to Gln did not affect the kinase activity whereas deletion of the ATP-binding domain eliminated it, indicating that the conserved His (H378) is not required for the kinase activity and this activity is intrinsic to the NTHK1-KD. Moreover, confocal analysis of NTHK1 expression in insect cells and plant cells suggested the plasma membrane localization of the NTHK1 protein. Thus, NTHK1 may represent a distinct Ser/Thr kinase-type ethylene receptor and function in an alternative mechanism for ethylene signal transduction. PMID- 12535352 TI - The role of invertases and hexose transporters in controlling sugar ratios in maternal and filial tissues of barley caryopses during early development. AB - To analyse carbohydrate metabolism and its role during early seed development of barley we characterised genes encoding two cell wall-bound invertases (HvCWINV1 and HvCWINV2) and two putative hexose transporter-like genes (HvSTP1 and HvSTP2). No typical vacuolar invertase gene could be identified. Instead, a gene encoding sucrose:fructan 6-fructosyltransferase (HvSF6FT1), an enzyme with soluble acid invertase activity, was isolated and characterised. Furthermore, enzyme activities and sugar levels were measured. HvSF6FT1-mRNA levels and acid soluble invertase activity are highest in the maternal pericarp 1-2 days after flowering (DAF). HvSF6FT1 is strongly expressed in regions flanking the main vascular bundle and to a lower extent in filial endospermal transfer cells, which persist until maturity and never accumulate starch. In contrast, cell wall-bound invertase HvCWINV2 is expressed early in development mainly in the style region and later on in pericarp areas which transiently accumulate starch and undergo degradation later in development. The hexose transporter HvSTP2 shows a temporal and spatial expression pattern similar to HvCWINV2. Transcripts of HvCWINV1 have been localised within the first row of endospermal cells and in the outermost area of the nucellar projection as well as in endospermal transfer cells before starch filling; the same regions of the endosperm are labelled with a hexose transporter HvSTP1-probe. HvSTP1 is expressed at very low levels within the pericarp but much higher in the syncytial endosperm at 3 DAF and in endospermal transfer cells 7 DAF. The temporal and spatial association of HvCWINV1 and HvSTP1 expression indicates that hexoses liberated by the invertase within the endospermal cavity are taken up by the transporter to be delivered into the central uncellularised space of the endosperm to supply mitotically active endosperm cells with hexoses. The results are discussed and compared with published data on the role of soluble sugars as signal molecules in seed developmental processes. PMID- 12535353 TI - The Arabidopsis ABORTED MICROSPORES (AMS) gene encodes a MYC class transcription factor. AB - Visual screening of a T-DNA mutagenised population of Arabidopsis thaliana for an absence of silique elongation lead to the isolation of the aborted microspores (ams) mutant that shows a sporophytic recessive male sterile phenotype. Homozygous mutant plants are completely devoid of mature pollen. Pollen degeneration occurs shortly after release of the microspores from the tetrad, prior to pollen mitosis I. Premature tapetum and microspore degeneration are the primary defects caused by this lesion, while a secondary effect is visualised in the stamen filaments, which are reduced in length and lie beneath the receptive stigma at flower opening. The disrupted gene was isolated and revealed a T-DNA element to be inserted into the eighth exon of a basic helix-loop-helix (bHLH) gene located on chromosome II. This protein sequence contains a basic DNA binding domain and two alpha helices separated by a loop, typical of a transcription factor belonging to the MYC sub family of bHLH genes. Therefore, AMS plays a crucial role in tapetal cell development and the post-meiotic transcriptional regulation of microspore development within the developing anther. PMID- 12535354 TI - Combined microsurgical and endovascular management of complex intracranial aneurysms. AB - OBJECTIVE: The disciplines of microneurosurgery and cranial base surgery have reached maturity, and technical advances in the surgical management of aneurysms are limited. Although most aneurysms can be clipped microsurgically or coiled endovascularly, a subset of patients may require a combined approach. A consecutive series of patients with aneurysms in one surgeon's cerebrovascular practice was reviewed retrospectively to analyze strategies for integrating microsurgical and endovascular techniques in the management of complex aneurysms. METHODS: Between 1997 and 2001, 596 aneurysms in 491 patients were treated microsurgically by the senior author (MTL) at the University of California, San Francisco, and 77 of these patients (96 aneurysms) were managed with a multimodality approach comprising a total of eight different combinations: selective revascularization and aneurysm occlusion (n = 23), endovascular and surgical trapping (n = 1), clipping of the aneurysm after attempted or incomplete coiling (n = 22), coiling after attempted or incomplete clipping (n = 5), clipping of recurrent aneurysm after coiling (n = 6), coiling of recurrent aneurysm after clipping (n = 1), clipping and coiling of multiple remote aneurysms (n = 13), and coiling after previous surgery (n = 6). RESULTS: A total of 96 aneurysms were treated with combined therapy, of which 43% were large or giant in size and 34% had fusiform or dolichoectatic morphology. Complete angiographic obliteration was achieved in 91 aneurysms (95%). Overall, 66 patients (86%) had good outcomes (Glasgow Outcome Scale score of 4 or 5; mean follow-up, 9 mo). The treatment mortality rate was 9.1% (seven patients), and permanent treatment-associated neurological morbidity rate was 5.2% (four patients). CONCLUSION: Evolving endovascular technologies need to be integrated into the microsurgical management of aneurysms. Multimodality approaches are best used with complex aneurysms in which conventional therapy with a single modality has failed. Revascularization remains a unique surgical contribution to the overall management of aneurysms with which current endovascular techniques cannot be used. Multimodality management should be considered an elegant addition to the therapeutic armamentarium that, through simplification and increased safety, improves the treatment of complex aneurysms beyond what is achievable by performing clipping or coiling alone. PMID- 12535355 TI - Hypomagnesemia after aneurysmal subarachnoid hemorrhage. AB - OBJECTIVE: Hypomagnesemia frequently occurs in hospitalized patients, and it is associated with poor outcome. We assessed the frequency and time distribution of hypomagnesemia after aneurysmal subarachnoid hemorrhage (SAH) and its relationship to the severity of SAH, delayed cerebral ischemia (DCI), and outcome after 3 months. METHODS: Serum magnesium was measured in 107 consecutive patients admitted within 48 hours after SAH. Hypomagnesemia (serum magnesium <0.70 mmol/L) at admission was related to clinical and initial computed tomographic characteristics by means of the Mann-Whitney U test. Hypomagnesemia at admission and during the DCI onset period (Days 2-12) was related to the occurrence of DCI and hypomagnesemia at admission, and hypomagnesemia that occurred any time during the first 3 weeks after SAH was related to outcome. RESULTS: Hypomagnesemia at admission was found in 41 patients (38%) and was associated with more cisternal (P = 0.006) and ventricular (P = 0.005) blood, a longer duration of unconsciousness (P = 0.007), and a worse World Federation of Neurosurgical Societies scale score at admission (P = 0.001). The crude hazard ratio for DCI with hypomagnesemia at admission was 2.4 (95% confidence interval, 1.0-5.6), and after multivariate adjustment it was 1.9 (95% confidence interval, 0.7-4.7). The hazard ratio of hypomagnesemia from Days 2 to 12 for patients with DCI was 3.2 (range, 1.1-8.9) after multivariate adjustment. The crude odds ratio for poor outcome (Glasgow Outcome Scale score, 1-3) with hypomagnesemia at admission was 2.5 (range, 1.1-5.5). Hypomagnesemia at admission did not contribute to the prediction of outcome in the multivariate model. CONCLUSION: Hypomagnesemia is frequently present after SAH and is associated with severity of SAH. Hypomagnesemia occurring between Days 2 and 12 after SAH predicts DCI. PMID- 12535356 TI - Neurosurgical management of intracranial aneurysms previously treated with endovascular therapy. AB - OBJECTIVE: With the increased use of endovascular therapy, an increasing number of patients with incompletely treated intracranial aneurysms are presenting for further surgical management. This study reviews our experiences with such patients. METHODS: During a 7-year period, 38 patients with 40 intracranial aneurysms who were initially treated with endovascular therapy underwent surgical obliteration of refractory or recurrent lesions. All patients were recorded in a prospective registry, and their clinical data and imaging studies were analyzed retrospectively. RESULTS: Twenty-six anterior and 14 posterior circulation aneurysms were treated. Four aneurysms were on the cavernous internal carotid artery, 13 were on the distal internal carotid artery, 6 were on the anterior communicating artery complex, 2 were on the middle cerebral artery, 3 were on the posteroinferior cerebellar artery, 1 was at the vertebrobasilar junction, 3 were on the superior cerebellar artery, 4 were at the basilar apex, 2 were on the posterior cerebral artery, and 1 was on the distal vertebral artery. Two pseudoaneurysms-one on the petrocavernous segment of the internal carotid artery and one on the distal VA-also were treated. The median time until recurrence was 6 months. Thirty-one aneurysms were clip-ligated, and six were treated with trapping. Three extracranial-intracranial bypasses were performed. One aneurysm was treated with muslin wrapping. Two aneurysms required the use of surgical approaches that involved hypothermic circulatory arrest. Nine aneurysms required coil mass extraction and/or complex vascular reconstruction to complete lesion obliteration. All aneurysms except the single wrapped aneurysm were successfully excluded from the intracranial circulation. Two deaths occurred as a result of the operative procedures, and another patient died as a result of subarachnoid hemorrhage-induced massive myocardial infarction. Ultimately, 86.8% of patients achieved an excellent or good recovery. CONCLUSION: With endovascular therapy assuming an increasing role in the treatment of patients with intracranial aneurysms, more lesions that are refractory to initial treatment will require surgical management. Our experience indicates that good results are attainable, although technical challenges are frequently encountered. PMID- 12535357 TI - Analysis of factors predictive of success or complications in arteriovenous malformation radiosurgery. AB - OBJECTIVE: This study was undertaken to determine which factors were statistically predictive of radiological and clinical outcomes in the radiosurgical treatment of arteriovenous malformations (AVMs). METHODS: The computerized dosimetric and clinical data for 269 patients were reviewed. The AVM nidus was hand-contoured on successive enhanced computed tomographic slices through the nidus, to allow detailed determinations of nidus volume, target miss, normal brain tissue treated, dose conformality, and dose gradient. In addition, a number of patient and treatment factors, including Spetzler-Martin grade, presenting symptoms, dose, number of isocenters, radiological outcome, and clinical outcome, were subjected to multivariate analysis. RESULTS: Two hundred twenty-five patients were treated with radiosurgery for the first time, and 44 patients underwent radiosurgical retreatment. One hundred forty-three patients had AVMs located in or near "eloquent" brain areas and 126 patients did not. Seventy patients demonstrated preoperative neurological findings related to the AVM and 199 did not. Twenty-six patients had previously undergone endovascular treatment and 10 patients had previously undergone surgical treatment of their AVMs. Of the 269 patients studied, 228 experienced no complication, 10 (3.7%) experienced a transient radiation-induced complication, 3 (1%) experienced a permanent radiation-induced complication, and 28 (10%) experienced posttreatment hemorrhage. CONCLUSION: None of the analyzed factors was predictive of hemorrhage after radiosurgery in this study. The 12-Gy volume was predictive of permanent radiation-induced complications. Eloquent AVM location and 12-Gy volume were correlated with the occurrence of transient radiation-induced complications. Better conformality was correlated with a reduced incidence of transient complications. Lower Spetzler-Martin grades, higher doses, and steeper dose gradients were correlated with radiological success. PMID- 12535358 TI - Radiation tolerance of functioning pituitary tissue in gamma knife surgery for pituitary adenomas. AB - OBJECTIVE: This study is intended to contribute to a determination of the sensitivity of preserved hypophyseal function to focal radiation in pituitary adenomas. METHODS: We compared two subgroups of patients followed up for a median of 5 years after gamma knife surgery (GKS). Subgroup 1 (n = 30) showed postirradiation hypopituitarism. Subgroup 2 (n = 33) was continually eupituitary. These subgroups were taken from a previously published study relating to a larger group of 163 patients with pituitary adenomas treated by GKS and evaluated after a median follow-up period of 2 years. A relatively high treatment dose was used in this larger group (median, 20 Gy to the tumor margin for growth control in nonfunctioning adenomas; median, 35 Gy for hypersecreting adenomas). Early results approached those of microsurgery, and there were only a few side effects. In the present study, we compared 16 different variables in the same two subgroups to discover the relationships that caused a delayed appearance of postirradiation hypopituitarism. The main pretreatment and treatment parameters were measured on reconstructed treatment plans. This database was used for statistical evaluation. RESULTS: The relationship between the mean dose and the volume of functioning hypophysis was stronger in terms of worsening of pituitary function than that of the spot dose to different intrasellar structures. We found that for our group of patients, the safe mean dose of radiation to the hypophysis was 15 Gy for gonadotropic and thyrotropic functions and 18 Gy for adrenocorticotropic function. The worsening of pituitary function was also significantly dependent on the dose to different anatomic levels of the infundibulum, but we did not succeed in fully characterizing this relationship. In addition, we discovered significant levels of dependency of postirradiation pituitary damage to different pretreatment and treatment variables. CONCLUSION: Knowledge of the radiation tolerance of functioning pituitary structures subjected to GKS can ensure better preservation of pituitary function after irradiation. This is valid for the group of patients we studied. Our study's findings can be used as a guideline for GKS treatment of new patients with pituitary adenomas, and it can serve for comparison with the experience of other gamma knife centers. PMID- 12535359 TI - Fibrous dysplasia of the clivus. AB - OBJECTIVE: Fibrous dysplasia is a developmental skeletal disorder that may lead to distortion, expansion, and weakening of the bone. Craniofacial involvement by this entity is well recognized and is known to cause neurovascular impingement and cosmetic deformity; fibrous dysplasia of the clivus, however, is unrecognized and seldom reported. Differentiating this entity from more aggressive disease processes affecting the clivus is central for the proper management of lesions in this area. We have studied fibrous dysplasia of the clivus with the goal of depicting its manifestations, outlining its management, and heightening awareness of this disease entity. METHODS: We retrospectively reviewed our database and identified patients with the diagnosis of fibrous dysplasia of the clivus. The demographic data, the clinical and radiological findings, and the management of these patients were reviewed. RESULTS: Eight patients who had experienced fibrous dysplasia of the clivus were identified. They either were asymptomatic (four patients) or presented with headache (four patients). Of the patients who presented with headache, one also had XIIth cranial nerve paralysis and another had dysphagia. The radiological findings for this entity were consistent, with typical findings of hypointensity on T1- and T2-weighted magnetic resonance imaging studies and ground-glass appearance on computed tomographic scans. Four of the patients in our series had pathological confirmation. Treatment was determined by the patient's symptoms. CONCLUSION: Fibrous dysplasia should be considered in the differential diagnosis of lesions affecting the clivus. Its clinical and radiological presentations permit the establishment of the diagnosis. Its management is usually conservative, unless the patient presents with nerve compression or extensive symptomatic involvement of the condyle. PMID- 12535360 TI - Combined-modality treatment of leptomeningeal gliomatosis. AB - OBJECTIVE: Leptomeningeal gliomatosis (LG) is a clinically uncommon metastatic complication of high-grade gliomas (HGGs), for which there is no consensus regarding treatment. The goal of this study was to determine the toxicity and response rate of combined-modality therapy for the treatment of patients with HGGs and LG. METHODS: Eighteen patients (10 men and 8 women), ranging in age from 28 to 70 years (median, 38 yr), with clinically, neuroradiologically, and cytologically documented LG received intraventricular chemotherapy. Tumor histological types included anaplastic astrocytoma (10 patients) and glioblastoma multiforme (8 patients). Concurrent radiotherapy (11 patients) or systemic chemotherapy (13 patients) was administered as clinically indicated. Methotrexate was administered initially, and treatment was continued for patients in stable or improved condition. For patients who experienced progression, cytosine arabinoside was administered as second-line therapy, followed by N,N',N" triethylenethiophosphoramide as third-line therapy. Patients underwent bimonthly evaluations with cerebrospinal fluid cytological assessments and neurological examinations. RESULTS: Four to 13 cycles (median, 5 cycles) of intraventricular chemotherapy were administered. Toxicity included aseptic meningitis (12 patients), radiation-induced enteritis (2 patients), and myelosuppression of Grade II or less (4 patients). No patient required hospitalization or transfusions, and no treatment-related deaths occurred. Partial responses were observed for 6 patients, and 12 patients demonstrated progressive disease. The median duration of response was 3 months (range, 2-4 mo). Survival times after the initiation of intraventricular chemotherapy ranged from 2 to 8 months (median, 3.5 mo). The cause of death was progressive LG (14 patients), combined LG and HGG (3 patients), and HGG (1 patient). CONCLUSION: For this small cohort of patients, combined-modality therapy had modest toxicity but minimal palliative efficacy. For the majority of patients with LG, supportive care should be considered. PMID- 12535361 TI - Surgical management and outcome of tuberculous atlantoaxial dislocation: a 15 year experience. AB - OBJECTIVE: Tuberculous atlantoaxial dislocation is a rare disease entity. However, tuberculosis continues to be endemic in developing countries. Its earliest clinical presentation may be nonspecific, and delay in diagnosis may lead to irreversible neurological deficit. The management of tuberculous atlantoaxial dislocation includes ventral cervicomedullary decompression, occipitocervical arthrodesis, and administration of antituberculous medications. METHODS: Eighteen patients with tuberculous atlantoaxial dislocation who presented with neck pain and/or occipital headache, restriction of neck movement, difficulty swallowing, and signs of myelopathy were studied. Four patients had evidence of associated pulmonary tuberculosis. Plain x-rays of the cervical spine, computed tomographic scans, and magnetic resonance images were obtained in all patients for diagnosis and to assess the degree of dislocation and cervicomedullary compression. Simultaneous anterior neural decompression, via a transcervical retropharyngeal approach, and posterior arthrodesis were performed on all patients while they remained under anesthesia. Antituberculous chemotherapy was continued for 18 months. RESULTS: Histopathological analysis of excised tissue was consistent with tuberculosis in all patients. However, Ziehl Neelsen staining for acid-fast bacilli was positive in two cases, and culture for Mycobacterium tuberculosis was negative in all patients. Patients with severe myelopathy experienced marked improvement. One patient died of fulminant resistant tuberculous meningitis. CONCLUSION: The transcervical retropharyngeal approach to the craniovertebral junction provides direct access to the lesion and avoids the potential bacterial contamination of the oral and pharyngeal cavity. It also prevents the development of persistent fistulae. Posterior stabilization should be performed directly after anterior neural decompression, while the patient remains under anesthesia, to prevent neurological deterioration before subsequent posterior fixation. This technique also is helpful for early mobilization of patients. The aim of surgical treatment should be to obtain biopsy tissue and to perform radical excision of epidural granulation tissue/abscess and infected bone using microsurgical technique. Antituberculous medication must be continued for 18 months with four drug regimens, and continuous monitoring of drug toxicity should be performed throughout the course of treatment. PMID- 12535362 TI - Comparison of cerebral blood flow in computed tomographic hypodense areas of the brain in head-injured patients. AB - OBJECTIVE: Hypodense lesions identified on computed tomographic (CT) scans are often assumed to indicate ischemia. The purpose of this study was to investigate regional cerebral blood flow (rCBF) in hypodense areas of the brain after severe traumatic brain injury. METHODS: CBF was measured by stable xenon-enhanced CT scans. Hypodense areas were identified, and rCBF values as well as CT density were averaged for the region. RESULTS: Thirty (60%) of the 50 patients had a total of 45 hypodense regions, which were associated with either contusion (n = 30) or areas of infarction (n = 15). rCBF in the hypodense regions was variable, ranging from a low of 3.3 to a high of 72.5 ml/100 g/min. The cause of the lesion was the major factor associated with the level of rCBF. Although the average decrease in CT density was similar for the two types of lesions, the average rCBF was significantly lower and the difference in rCBF between the lesion and the contralateral side was greater when the hypodense lesion was associated with a contusion. A critical reduction in rCBF (<20 ml/100 g/min) was found in 19 (63%) of the hypodense regions associated with contusions but in only 4 (27%) of those from areas of infarction. CONCLUSION: Hypodensity on plain CT scans does not always indicate reduction in CBF. This association was found more commonly when the low-density area was associated with a contusion. In hypodense areas associated with infarction, rCBF was variable and not commonly in the ischemic range at the time the CBF measurement was obtained. PMID- 12535363 TI - Shunt revision for asymptomatic failure: surgical and clinical results. AB - OBJECTIVE: Shunt obstruction represents a permanent threat for patients with shunts, and its prevention and treatment are important parts of neurosurgeons' duties. Although there is little discussion regarding the need to reoperate for treatment of symptomatic shunt failure (SSF), the need to reoperate for treatment of asymptomatic shunt failure (ASF) is debated, as are the guidelines for the follow-up monitoring of patients with shunts. The goal of this study was to assess the effects of systematic follow-up monitoring and shunt revision for ASF; we reviewed our database to compare the results of shunt revision for ASF versus SSF. METHODS: We defined ASF as shunt failure diagnosed for an asymptomatic patient during a systematic consultation. In our institution, children who receive shunts for treatment of hydrocephalus are systematically monitored in an outpatient clinic, with clinical examinations and plain x-rays. Among 1,564 children with shunts, who were monitored for a mean of 10.7 years, 1106 (70.7%) required at least one shunt revision. The indication for the first revision was SSF in 609 cases and ASF in 305 cases; the indication was not specified in 192 cases. We studied the surgical outcomes after the first shunt revision and compared the results for SSF and ASF. RESULTS: After the first revision, shunt infections and subsequent shunt failure were significantly less frequent in the ASF group, compared with the SSF group. The interval between the first shunt revision and subsequent shunt failure was significantly longer in the ASF group. CONCLUSION: Our data support the practice of systematic follow-up monitoring for patients with shunts, for the early diagnosis and systematic treatment of ASF. PMID- 12535364 TI - Familial aggregation of intracranial aneurysms in an Inuit patient population in Kalaallit Nunaat (Greenland). AB - OBJECTIVE: The incidence of subarachnoid hemorrhage (SAH) and intracranial aneurysm (IA) has been reported to be higher in Greenlandic Inuits than in Caucasian Danes, but the rate of familial aggregation in Inuits is unknown. METHODS: This study retrospectively compared the rate of familial aggregation of SAH and IA (at least one first- or second-degree relative with presumed SAH and/or IA) in 120 Inuit patients from Greenland admitted to the Copenhagen University Hospital in Copenhagen, Denmark, from 1978 to 1998 with a diagnosis of ruptured IA with that in 1,037 Caucasian Danes admitted from 1978 to 1983. RESULTS: Inuit patients had a much higher rate of familial history of SAH (23.1%) and of IA (9.6%) than Danish patients (4.3 and 1.6%, respectively). In both populations, familial SAH was associated with lower age at the time of aneurysm rupture. Danish patients with familial SAH showed a higher rate of middle cerebral artery aneurysms (40 versus 26% in sporadic SAH). In Inuit patients with familial and nonfamilial SAH, 42 and 38% of the aneurysms originated from the middle cerebral artery. The overall rate of multiple aneurysms was highest among Inuits, and in both populations, it was increased in the presence of a positive family history. CONCLUSION: The rate of a positive family history of presumed SAH and IA is high among Inuits who present with SAH compared with Caucasian Danes who present with SAH. This finding, coupled with a higher rate of multiple aneurysms and younger age at presentation, suggests a potential genetic influence among Inuit families. PMID- 12535365 TI - Far posterior subtemporal approach to the dorsolateral brainstem and tentorial ring: technique and clinical experience. AB - OBJECTIVE: Surgical access to the posterolateral mesencephalon or tentorial ring can be problematic, depending on the angle of the tentorium and associated venous structures. A far posterior subtemporal approach was developed that uses the wide opening of the tentorium and the option of supratentorial retraction of the cerebellum to provide an excellent angle of approach to this region. Details of this technique and a series of eight patients with lesions treated by this approach are presented. METHODS: Seven cavernous malformations and one dural arteriovenous malformation were approached in eight patients. In this far posterior subtemporal approach, a horseshoe incision is centered slightly behind the ear, with the caudal extent of the craniotomy defined by the temporal fossa floor. The entry of the vein of Labbe into transverse sinus is identified intradurally. If the vein enters anteriorly in the exposure, it is mobilized. Retractors are then placed, and the tentorium is visualized and opened laterally, avoiding the trochlear nerve. A thin, tapered retractor can be used to retract the cerebellum posterolaterally away from the brainstem. RESULTS: This approach was used to treat eight patients, and their lesions were successfully resected. Seven patients experienced good outcomes, and one patient, who presented with Hunt-Hess Grade IV subarachnoid hemorrhage from a dural arteriovenous malformation, experienced a fair outcome. CONCLUSION: The far posterior subtemporal approach is effective for approaching carefully selected lesions of the posterolateral mesencephalon and tentorial ring. PMID- 12535366 TI - Minimally invasive cervical expansile laminoplasty: an initial cadaveric study. AB - OBJECTIVE: Expansile laminoplasty has been successfully used to treat cervical myelopathy attributable to canal stenosis. However, detachment of the posterior cervical muscles is thought to contribute to postoperative axial neck pain and kyphosis. Minimizing the amount of muscular dissection might reduce the likelihood of these sequelae. METHODS: Six human cadaveric spines were used to assess the feasibility of a minimally invasive laminoplasty technique. A 22-mm tubular dilator port was used to access the lamina-facet junctions from C2 to C7, through bilateral stab incisions at C4-C5 and C5-C6. Troughs at the lamina-facet junctions were drilled bilaterally, and the contiguous laminae were lifted en bloc from one side. Ten-millimeter rib allograft spacers were inserted to maintain a gap on the open side. RESULTS: Exposure of six cervical levels was easily accomplished with two small incisions on each side. Drilling was achieved without dural violations. The midsagittal spinal canal diameter was increased by a mean of 38% and the spinal canal area was increased by an average of 43% at the level of C5. CONCLUSION: A minimally invasive approach for cervical laminoplasty could be performed in human cadavers. The measured increases in spinal canal space approximated those demonstrated to be associated with stabilization or improvement of neurological status. PMID- 12535367 TI - Endoscopic endonasal approach to the pterygopalatine fossa: anatomic study. AB - OBJECTIVE: The pterygopalatine fossa is a relatively small anatomic region. Because of its rich vasculonervous contents and its connections with several intracranial and extracranial compartments, it is of particular surgical interest. Because of its deep localization and despite its small size, however, it can require extensive anatomic approaches, especially for invasive cranial base lesions. We performed a cadaveric study through a minimally invasive endoscopic endonasal approach to the pterygopalatine fossa. METHODS: We studied 16 pterygopalatine fossae in eight adult cadaveric heads in which the arteries and veins were injected with latex. For visualization, we used rod-lens endoscopes, 4 mm in diameter and 18 cm in length, with 0-, 30-, 45-, and 70 degree lenses. An endonasal middle meatal transpalatine approach, an endonasal middle meatal transantral approach, and an endonasal inferior turbinectomy transantral approach were used. RESULTS: The middle meatal transpalatine approach allows for medial exposure of the pterygopalatine fossa contents, the middle meatal transantral approach allows a lateral view, and the inferior turbinectomy transantral approach allows the widest view and room for surgical maneuvering in the medial and lateral compartments of the pterygopalatine fossa and the infratemporal fossa. CONCLUSION: Our anatomic study shows that this approach can be considered a valid minimally invasive option to approach pterygopalatine fossa lesions. PMID- 12535368 TI - Intraneural colchicine inhibition of adenoviral and adeno-associated viral vector remote spinal cord gene delivery. AB - OBJECTIVE: The mechanism of remote viral gene delivery to the spinal cord is unknown. The present experiment demonstrates that intraneural injection of colchicine is capable of inhibiting remote delivery of both adenoviral and adeno associated viral (AAV) vectors, implicating axonal transport in this process. METHODS: The right sciatic nerves of adult Sprague-Dawley rats were injected with phosphate-buffered saline (PBS) (n = 5) or 10 (n = 7) or 100 (n = 4) microg colchicine. Two days later, the nerves of all animals were initially injected with 1.2 x 10(7) plaque-forming units of Ad5RSVntLac-Z. Two separate groups were injected concurrently with vector and PBS (n = 5) or 10 microg colchicine (n = 5). In a second experiment, the right sciatic nerves of CD1 mice were preinjected with PBS (n = 6) or 10 microg colchicine (n = 5). Two days later, the nerves were injected with rAAVCAG-EGFPwpre (an adeno-associated vector carrying the green fluorescent protein gene). In both experiments, sciatic nerves and spinal cords were removed and analyzed for gene expression. RESULTS: Sciatic nerve vector injection resulted in expression in both the nerve injection site and neuronal cell bodies located predominantly in the ipsilateral ventral horn. Analysis of variance revealed a significant treatment effect for 10 and 100 microg intraneural colchicine with inhibition of remote adenoviral delivery at 10 microg and blockade of remote delivery at 100 microg (P < 0.001). Colchicine injection concurrent with and before vector injection had similar inhibitory effects. Two way analysis of variance revealed significant colchicine inhibition of remote delivery in both adenovirus- and AAV-injected animals (P < 0.003) but no dose-by vector interaction, suggesting that both vectors are equally inhibited by colchicine. CONCLUSION: Colchicine inhibits remote spinal cord delivery of adeno associated and adenoviral vectors in a dose-dependent manner, suggesting that remote delivery is dependent on retrograde axonal transport. PMID- 12535369 TI - Prolonged convection-enhanced delivery into the rat brainstem. AB - OBJECTIVE: Prolonged convection-enhanced delivery was used in an attempt to achieve large volumes of distribution (V(d)) in the rat brainstem. Clinical assessment and histological analysis were performed to establish the safety of this approach. METHODS: For evaluation of V(d,), 10 rats underwent stereotactic cannula placement into the brainstem. Five rats underwent a 24-hour infusion (volume of infusion [V(i)], 200 microl), and 5 rats underwent a 7-day infusion (V(i), 2 ml) of fluorescein isothiocyanate-dextran. Serial brainstem sections were imaged with ultraviolet illumination, and V(d) was assessed. For assessment of clinical tolerance, 30 additional rats underwent chronic infusions of an isotonic saline solution into the brainstem. Serial neurological examinations were performed, followed by histological analysis after the animals' death. RESULTS: No animal demonstrated clinically recognized neurological deficits. Foci of organizing necrosis were limited to the site of infusion and cannula tract. V(d) increased linearly with increasing V(i) (range, 24.8-130.6 mm(3)). Maximal cross sectional area of fluorescence and craniocaudal extent of fluorescence increased with increasing V(i). Fluorescence was detected throughout the entire brainstem beyond the compact area of highly concentrated tracer. CONCLUSION: Prolonged convection-enhanced delivery can be applied safely in the rat brainstem with no recognized limitations of V(d) and minimal histological changes beyond the site of infusion. Chronic brainstem infusions may enhance the potential application of convection-enhanced delivery for therapeutic purposes in treating diffuse pontine gliomas. PMID- 12535370 TI - Protective effect of C1 esterase inhibitor on reperfusion injury in the rat middle cerebral artery occlusion model. AB - OBJECTIVE: The complement system is thought to play a major role in initiating some of the inflammatory events that occur during reperfusion injury. The aim of this study was to assess the effects of C1 esterase inhibitor (C1-INH) on ischemic injury in the rat model of middle cerebral artery suture occlusion and reperfusion. METHODS: Thirty-six male Wistar rats were used. Intraluminal middle cerebral artery occlusion was performed for 60 minutes. Just before reperfusion, C1-INH (50 international units/kg) (C1-INH group, n = 19) or saline solution (control group, n = 17) was administered. Physiological parameters (arterial blood gas values, mean arterial blood pressure, and heart rate) and local cerebral blood flow were recorded during the experiment. Forty-eight hours after reperfusion, all rats were killed, and assessments of leukocyte infiltration with a myeloperoxidase activity assay and histological analyses with 2,3,5-triphenyl tetrazolium chloride staining were performed. RESULTS: The physiological parameters and local cerebral blood flow values were not significantly different in the two groups. The infarction volume was significantly smaller and the myeloperoxidase activity was significantly lower in the C1-INH group (84.9 +/- 69.1 mm(3) and 0.40 +/- 0.29 units/g, respectively) than in the control group (202.3 +/- 98.3 mm(3) and 1.41 +/- 0.44 units/g, respectively) (P < 0.01). Myeloperoxidase activities were strongly correlated with infarction volumes (r = 0.73, P < 0.01). CONCLUSION: The results of this study indicated that C1-INH reduced polymorphonuclear leukocyte accumulation and neuronal damage in focal ischemia and reperfusion. PMID- 12535371 TI - Endovascular restorative neurosurgery: a novel concept for molecular and cellular therapy of the nervous system. AB - The amalgam of molecular biology and neurosurgery offers immense promise for neurorestoration and the management of neurodegenerative deficiencies, developmental disorders, neoplasms, stroke, and trauma. This article summarizes present strategies for and impediments to gene therapy and stem cell therapy of the central nervous system and advances the concept of a potential new approach, namely endovascular restorative neurosurgery. The objectives of gene transfer to the central nervous system are efficient transfection of host cells, selective sustained expression of the transgene, and lack of toxicity or immune excitation. The requisite elements of this process are the identification of candidate diseases, the construction of vehicles for gene transfer, regulated expression, and physical delivery. In the selection of target disorders, the underlying genetic events to be overcome, as well as their spatial and temporal distributions, must be considered. These factors determine the requirements for the physical dispersal of the transgene, the duration of transgene expression, and the quantity of transgene product needed to abrogate the disease phenotype. Vehicles for conveying the transgene to the central nervous system include viral vectors (retroviruses, lentiviruses, adenoviruses, adeno-associated viruses, and herpes simplex virus), liposomes, and genetically engineered cells, including neural stem cells. Delivery of the transgene into the brain presents several challenges, including limited and potentially risky access through the cranium, sensitivity to volumetric changes, restricted diffusion, and the blood-brain barrier. Genetic or cellular therapeutic agents may be injected directly into the brain parenchyma (via stereotaxy or craniotomy), into the cerebrospinal fluid (in the ventricles or cisterns), or into the bloodstream (intravenously or intra arterially). The advantages of the endovascular route include the potential for widespread distribution, the ability to deliver large volumes, limited perturbation of neural tissue, and the feasibility of repeated administration. PMID- 12535372 TI - The Charcot library. AB - The Charcot library was officially created at the Salpetriere Hospital in 1907 after the donation of the private library of Jean-Martin Charcot, which Dr. Charcot had accumulated progressively throughout his professional career. Increased by several other endowments (the most important being the collection of the Resident's Library, begun in 1886) and other private donations, the library became officially affiliated with the Paris VI University in 1985. It now functions as an entity oriented toward the neurosciences and offers a large number of 19th-century books (including all of Charcot's manuscripts) and more recent documents. Digitization of the most classic holdings, including atlases, will soon make them accessible through the library's web site. PMID- 12535373 TI - First Turkish neurosurgical journal (1936-1947): Modern Cerrahi ve norosirurji mecmuasi. AB - The publication of neurosurgery-related journals began in the late 1920s. Most of these journals are of Estonian, German, English, American, and French origin. The appearance of such journals is related to the development of neurosurgery and the number of neurosurgeons. This study presents a neurosurgical journal, Modern Cerrahi ve norosirurji mecmuasi (Journal of Modern Surgery and Neurosurgery), published between 1936 and 1947 in Turkey. The contents of the journal and some selected articles are also presented. The existence of this journal, which was published before most Western neurosurgical journals, should be regarded from a historical point of view. PMID- 12535374 TI - Research progress in the last quarter of the 20th century at the University of Tokyo and Tokyo Women's Medical University. AB - Professor Keiji Sano described the history of neurosurgery in Japan until 1975. After World War II, not only neurosurgery but all fields of medicine were devastated in Japan. Professor Sano contributed greatly to the reform and modernization of neurosurgery during that very difficult era in Japan. He performed much research by himself and also as a leader of research groups on stereotactic and functional neurosurgery, cerebrovascular diseases, head injuries, and brain tumors. He organized the Fifth International Congress of Neurological Surgery in Tokyo in 1973. I succeeded in the chairmanship of the Department of Neurosurgery of the University of Tokyo in 1981. We have performed research on the treatment of brain tumors and cerebrovascular diseases. To obtain the best results for brain tumor treatment, we have introduced several new radiotherapeutic methods, such as the gamma knife, heavy-particle irradiation, and the photon radiosurgery system. To improve surgical treatment, we have energetically engaged in medical engineering research on computer-assisted surgical systems (intraoperative monitoring and navigation systems). We have also performed much research on chemotherapy and immunotherapy. In the field of cerebrovascular diseases, the main research projects have been focused on the mechanism and treatment of vasospasm and brain edema after subarachnoid hemorrhage. I summarize the results of our research performed in the Department of Neurosurgery of the University of Tokyo until 1992 and at Tokyo Women's Medical University after 1992, in the last quarter of the 20th century. PMID- 12535375 TI - Internal carotid plexus schwannoma of the cavernous sinus: case report. AB - OBJECTIVE AND IMPORTANCE: Schwannomas of the central nervous system usually originate from the vestibular nerve and occasionally originate from the trigeminal nerve. Sympathetic plexus schwannomas are extremely rare and have never been noted within the cavernous sinus. CLINICAL PRESENTATION: A 23-year-old man experienced occasional double vision for a period of 6 months. Magnetic resonance imaging studies revealed an isointense lesion, with enhancement after gadolinium administration, located inferomedial to the internal carotid artery within the left cavernous sinus. INTERVENTION: We explored the cavernous sinus via a left-sided extradural-pterional approach and found the tumor inferomedial to the cavernous segment of the internal carotid artery. Microsurgical gross total resection of the tumor was performed. The IIIrd (oculomotor) to VIth (abducens) cranial nerves within the cavernous sinus were not related to the tumor and were preserved. The operative findings and the anatomic location of the tumor demonstrated that it originated from the internal carotid plexus within the cavernous sinus. The patient's postoperative course was uneventful, and he exhibited no cranial nerve deficits. However, incomplete Horner's syndrome was present on the treated side. CONCLUSION: We present the first reported case of an internal carotid plexus schwannoma, and we describe in detail its anatomic and neuroradiological characteristics. The microneurosurgical resection of this unusual tumor within the cavernous sinus was successful and without morbidity. PMID- 12535376 TI - Scalp arteriovenous malformation draining into the superior sagittal sinus associated with an intracranial arteriovenous malformation: just a coincidence? Case report. AB - OBJECTIVE AND IMPORTANCE: Recent experimental and clinical evidence suggests that hemodynamic changes in the venous system can induce the formation of new arteriovenous malformations (AVMs). In a rat model, increased venous pressure induces the formation of soft tissue and dural AVMs. We report a clinical observation that may support these data. CLINICAL PRESENTATION: A 4-year-old boy with a midline scalp AVM draining into the superior sagittal sinus had an associated intracranial/parenchymal AVM. The cerebral AVM increased venous pressure in the superior sagittal sinus as revealed by angiography. INTERVENTION: The scalp AVM was resected, and the intracranial AVM was treated by use of the gamma knife. CONCLUSION: On the basis of reported experimental data and the morphological and hemodynamic characteristics in this patient's two lesions, we suggest that the scalp AVM might have been induced by hypertension in the superior sagittal sinus. This clinical observation supports the notion suggested by experimental studies that hemodynamic changes can induce the formation of associated AVMs. PMID- 12535377 TI - Minimally invasive lateral mass screws in the treatment of cervical facet dislocations: technical note. AB - OBJECTIVE: The technique of lateral mass screw and rod or plate fixation is a major advancement in the posterior instrumentation of the cervical spine. This technique provides rigid three-dimensional fixation, restores the dorsal tension band, and provides highly effective stabilization in patients with many types of traumatic injuries. METHODS: Patient 1 was a 32-year-old man who had been in a motor vehicle accident. He presented with right C5 radiculopathy. X-ray findings included 45% anterolisthesis of C4 on C5, bilateral facet disruption, and right unilateral C4-C5 facet fracture and dislocation. The patient was placed in Gardner-Wells tongs, and the fracture was reduced with 25 pounds of traction. Patient 2 was a 56-year-old woman who had been in a motor vehicle accident that resulted in complete quadriplegia. Her initial imaging studies revealed a C3-C4 right unilateral facet fracture with subluxation. She was placed in traction, and her neurological status was reassessed. The findings of her neurological examination revealed improvement: she was found to have Brown-Sequard syndrome. Patient 3 was a 33-year-old man who was involved in a diving accident that resulted in bilaterally jumped facets at C3-C4. The patient was neurologically intact, and attempts at closed reduction were not successful. RESULTS: Patients 1 and 2 underwent anterior cervical discectomy with iliac crest autograft fusion and plating. They were then placed in the prone position, and a dilator tubular retractor system was used to access the facet joint at the level of interest. The facet joints were then denuded and packed with autograft. Lateral mass screws were then placed by means of the Magerl technique, and a rod was used to connect the top-loading screws. Patient 3 underwent posterior surgery that included only removal of the superior facet, intraoperative reduction, and bilateral lateral mass screw and rod placement. CONCLUSION: This technical note describes the successful placement of lateral mass screw and rod constructs with the use of a minimally invasive approach by means of a tubular dilator retractor system. This approach preserves the integrity of the muscles and ligaments that maintain the posterior tension band of the cervical spine. PMID- 12535378 TI - Vertebral autograft used as bone transplant for anterior cervical corpectomy: technical note. AB - OBJECTIVE: In this prospective patient study, we used a surgical technique for autograft bone fusion during anterior cervical corpectomy (ACC) in patients experiencing cervical spondylotic myelopathy. We packed the resected bone material of the corpectomy into a titanium mesh cage. To evaluate the efficacy of our autograft technique, we analyzed the results according to neurological outcome, radiological outcome, and complications. METHODS: Between 1995 and 1998, 27 ACC operations were performed for cervical spondylotic myelopathy caused by multisegmental cervical spondylosis. In all patients, decompression of the cervical canal and/or spinal nerve roots was performed by a median cervical corpectomy by an anterior approach. After the ACC was completed, a titanium mesh cage, which was variable in diameter and length, was filled with morselized and impacted bone material from the cervical corpectomy and was then implanted. An anterior cervical plate was placed in all patients to achieve primary stability of the cervical vertebral column. Age, sex, pre- and postoperative myelopathy, number of decompressed levels, radiological results, and complications were assessed. The severity of myelopathy was graded according to the scoring system of the Japanese Orthopaedic Association. RESULTS: Symptomatic improvement of neurological deficits was achieved in 80% of the patients. The mean preoperative Japanese Orthopaedic Association score improved from 13.1 to 15.2 postoperatively (P < 0.05). No patient demonstrated worsening of myelopathic symptoms. Radiological follow-up studies demonstrated complete bony fusion in all patients. A vertical movement of 2.25 +/- 0.43 mm of the titanium cage into the adjacent vertebral bodies was observed in 24 patients. In patients with either a lordotic or neutral cervical spinal axis postoperatively, the axis remained unchanged during the entire follow-up period. CONCLUSION: The results of this study demonstrate that transplantation of autograft bone material harvested during the ACC integrated well in the cage and in the adjacent vertebral bodies. Thus, complications associated with explantation of autograft material from other donor sites, e.g., the iliac crest, could be avoided. The early postoperative and midterm follow-up periods provided no evidence of morphological or functional instability of the operated cervical segments when this autograft technique was used in combination with cervical instrumentation. PMID- 12535379 TI - Internal carotid artery rotation technique to expose an aneurysm neck: technical note. AB - OBJECTIVE: Full exposure of a neck of an aneurysm is important to obliterate it completely. METHODS: A 51-year-old woman in whom an unruptured aneurysm projecting medially under the optic nerve was incidentally found underwent clipping surgery. The edge of the circumferentially cut distal dural ring was pulled with forceps to rotate the internal carotid artery and to expose the neck of the aneurysm. RESULTS: The neck of the aneurysm was completely obliterated under direct vision. CONCLUSION: This single, neat method exposes the neck of an unruptured paraclinoid aneurysm of the internal carotid artery. PMID- 12535380 TI - Successful transarterial Guglielmi detachable coil embolization of posttraumatic posterior communicating artery-cavernous sinus fistula: technical note. AB - OBJECTIVE: Carotid-cavernous fistulae are uncommon but well-documented sequelae of craniofacial trauma. A rare subset may arise from the posterior communicating artery instead of from the carotid artery proper. The presentation is similar to that of carotid-cavernous fistulae, with ocular pain, chemosis, and proptosis being the common symptoms. The first successful transarterial coil embolization of this type of lesion is described. METHODS: A 42-year-old man presented with severe craniocerebral injury, including multiple craniofacial fractures, after an industrial accident. He required emergent craniotomy for an open depressed cranial fracture and epidural hematoma. Six weeks after presentation, the patient began to exhibit progressive chemosis and proptosis. Magnetic resonance imaging revealed findings consistent with a carotid-cavernous fistula. RESULTS: Angiography revealed a fistula between the posterior communicating artery and the cavernous sinus. The origin of the fistula in the posterior communicating artery was successfully obliterated with Guglielmi detachable coiling. Subsequent studies demonstrated no flow through the fistula and good opacification of the ipsilateral posterior cerebral artery by the vertebrobasilar system. CONCLUSION: Posterior communicating artery-cavernous fistulae are a rare sequel of trauma. They may be treated successfully with the use of transarterial coil embolization. PMID- 12535381 TI - Robot-assisted thoracoscopic resection of a benign mediastinal neurogenic tumor: technical note. AB - OBJECTIVE: Robotic surgery systems were introduced recently with the objective of enhancing the dexterity and view during procedures that use a videoscope. The first case report of robot-assisted thoracoscopic removal of a benign neurogenic tumor in the thorax is presented. METHODS: A 46-year-old woman presented with a history of paravertebral pain. A chest x-ray revealed a left paravertebral mass. A magnetic resonance imaging scan revealed a well-encapsulated mass that was suspected to be a neuroma at the level of T8-T9, separate from vascular structures, without extension in the foramina, and without a spinal canal component. RESULTS: A left robot-assisted thoracoscopic resection of the tumor was performed. After placement of six trocars, the tumor was carefully dissected and removed through one of the trocar openings. The histopathological findings revealed an ancient schwannoma. CONCLUSION: This case report demonstrates the feasibility of robot-assisted thoracoscopic extirpation of a thoracic neurogenic tumor. Robot-assisted surgery may prove to be of additional value in challenging thoracoscopic surgery, such as the delicate surgical removal of benign neurogenic tumors, because of the support in manipulation and visualization during videoscopic interventions. PMID- 12535383 TI - Elective high frequency oscillatory ventilation versus conventional ventilation for acute pulmonary dysfunction in preterm infants. AB - BACKGROUND: Respiratory failure due to lung immaturity is a major cause of mortality in preterm infants. Although intermittent positive pressure ventilation (IPPV) saves lives, lung distortion during its use is associated with lung injury and chronic lung disease (CLD). Conventional IPPV is provided at 30-80 breaths per minute while a newer form of ventilation called high frequency oscillatory ventilation (HFOV) provides 'breaths' at 10-15 cycles per second. This has been shown to result in less lung injury in experimental studies. OBJECTIVES: The objective of this review is to determine whether the elective use of high frequency oscillatory ventilation as compared to conventional ventilation (CV) in preterm infants who are mechanically ventilated for the respiratory distress syndrome decreases the incidence of chronic lung disease, without adverse effects. SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal Trials, MEDLINE, EMBASE, previous reviews including cross references, abstracts, conferences and symposia proceedings, expert informants, journal hand searching by the Cochrane Collaboration, mainly in the English language. The search was updated in October 2002. SELECTION CRITERIA: Randomized controlled trials comparing HFOV and CV in preterm or low birth weight infants with pulmonary dysfunction, mainly due to RDS, who are to be given IPPV. Randomization and commencement of treatment should have been as soon as possible after the start of IPPV and usually in the first 12 hours of life. DATA COLLECTION AND ANALYSIS: The methodological quality of each trial was independently reviewed by the various authors. Each author extracted data separately; they were compared and differences were resolved. Treatment effects were expressed using relative risk (RR) and risk difference (RD). From 1/RD the number needed to treat (NNT) for benefits, and number needed to harm (NNH) for adverse effects, were calculated. 95% confidence intervals were used. MAIN RESULTS: Meta-analysis of the ten eligible studies comparing HFOV with CV revealed no evidence of effect on mortality at 28-30 days of age or at approximately term equivalent age. These results were consistent across studies. HFOV caused a significant reduction in CLD in survivors at term equivalent GA. However, the effect was not large in absolute terms [NNT 17 (10, 50)], and was inconsistent across studies. HFOV caused a significant reduction in the aggregated outcome, death or CLD at term equivalent GA. Again, however, the effect was not large [(NNT 20 (11, 100)] and was not consistent across studies. Pre-specified subgroup analyses according to use of a high volume strategy, or use of surfactant, did not identify subgroups in which there was evidence of effect on death, or in which the size of effect on CLD was substantially increased, or in which heterogeneity of treatment effect on CLD was removed. Short term neurological morbidity caused by HFOV was found in some studies, but this effect was not statistically significant overall. The subgroup of two trials not using a high volume strategy with HFOV found increased rates of Grade 3 or 4 IVH and of periventricular leukomalacia. An adverse effect of HFOV on longer term neurodevelopment was found in one large trial but not in two other small trials which reported this outcome. REVIEWER'S CONCLUSIONS: There is strong evidence that HFOV had no significant effect on death as this result was consistent across trials. Although HFOV caused a modest reduction in CLD, this evidence is weaker, because of inconsistencies across trials in this effect. In general, subgroup analyses according to use of high volume strategy, or surfactant, did not remove this heterogeneity, which therefore remains largely unexplained. Any future trials on elective HFOV should target those infants who are at most risk of CLD (extremely preterm), compare different strategies for generating HFOV and report important long term pulmonary and neurodevelopmental outcomes. Economic analysis should also be incorporated. PMID- 12535382 TI - Differential gene expression in human cerebrovascular malformations. AB - OBJECTIVE: We sought to identify genes with differential expression in cerebral cavernous malformations (CCMs), arteriovenous malformations (AVMs), and control superficial temporal arteries (STAs) and to confirm differential expression of genes previously implicated in the pathobiology of these lesions. METHODS: Total ribonucleic acid was isolated from four CCM, four AVM, and three STA surgical specimens and used to quantify lesion-specific messenger ribonucleic acid expression levels on human gene arrays. Data were analyzed with the use of two separate methodologies: gene discovery and confirmation analysis. RESULTS: The gene discovery method identified 42 genes that were significantly up-regulated and 36 genes that were significantly down-regulated in CCMs as compared with AVMs and STAs (P = 0.006). Similarly, 48 genes were significantly up-regulated and 59 genes were significantly down-regulated in AVMs as compared with CCMs and STAs (P = 0.006). The confirmation analysis showed significant differential expression (P < 0.05) in 11 of 15 genes (angiogenesis factors, receptors, and structural proteins) that previously had been reported to be expressed differentially in CCMs and AVMs in immunohistochemical analysis. CONCLUSION: We identify numerous genes that are differentially expressed in CCMs and AVMs and correlate expression with the immunohistochemistry of genes implicated in cerebrovascular malformations. In future efforts, we will aim to confirm candidate genes specifically related to the pathobiology of cerebrovascular malformations and determine their biological systems and mechanistic relevance. PMID- 12535384 TI - Hormones for suspected impaired fetal growth. AB - BACKGROUND: It has been suggested that oestrogens may improve fetal growth due to an increase in nutritional supply to the fetus from greater uterine blood flow. OBJECTIVES: The objective of this review was to assess the effects of hormone administration for suspected impaired fetal growth and perinatal outcome. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register (1 November 2002). SELECTION CRITERIA: Acceptably controlled trials of hormone administration for suspected impaired fetal growth which report fetal, perinatal or maternal outcomes. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed. MAIN RESULTS: No studies were included since none of the potentially relevant trials reported clinical outcomes. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the clinical use of hormone administration for suspected impaired fetal growth. PMID- 12535385 TI - Immunotherapy for recurrent miscarriage. AB - BACKGROUND: Immunologic aberrations might be the cause of recurrent early pregnancy loss in some women. OBJECTIVES: The objective of this review was to assess the effects of leukocyte immunisation or other immunologic treatments such as intravenous immune globulin (IVIG) on the live birth rate in women with previous unexplained recurrent miscarriages. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched (September 2002). Individual patient data were obtained from the American Society for Reproductive Immunology. SELECTION CRITERIA: Randomised trials of immunotherapy in women with three or more prior miscarriages and no more than one live birth; all recognised non-immunologic causes ruled out and no simultaneous treatment intervention. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: Nineteen trials of high quality were included. The various forms of immunotherapy did not show significant differences between treatment and control groups in terms of subsequent live births: paternal cell immunisation (11 trials, 596 women), odds ratio (OR) 1.05, 95% confidence intervals (CI) 0.75 to 1.47; third party donor cell immunization (3 trials, 156 women), OR 1.39, 95% CI 0.68 to 2.82; trophoblast membrane infusion (1 trial, 37 women), OR 0.40, 95% CI 0.11 to 1.45; intravenous immune globulin (IVIG), OR 0.98, 95% CI 0.61 to 1.58. REVIEWER'S CONCLUSIONS: Paternal cell immunization, third party donor leukocytes, trophoblast membranes, and intravenous immune globulin provide no significant beneficial effect over placebo in preventing further miscarriages. PMID- 12535386 TI - Instruments for chorionic villus sampling for prenatal diagnosis. AB - BACKGROUND: Chorionic villus sampling (CVS) is the method of choice for obtaining fetal tissue for prenatal diagnosis before 15 weeks of pregnancy. CVS can be performed using either transabdominal or transcervical approach. The type of instrument used could have a significant impact on the success rate of the procedure. An ability to manoeuvre the instrument within the uterine cavity without puncturing the gestational sac, and to see the tip of the instrument on ultrasound scanning are particularly important. OBJECTIVES: The objective of this review was to assess the effects of instruments used to obtain chorionic tissue in early pregnancy by transabdominal or transcervical route (chorionic villus sampling). The outcomes of interest were technical difficulties during the procedure, quality and quantity of obtained tissue, maternal adverse effects, pregnancy outcome and cost-effectiveness. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. Date of last search: November 2002. SELECTION CRITERIA: Randomised trials comparing different instruments (forceps, cannula, needle) for chorionic villus sampling using either transabdominal or transcervical approach. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by two reviewers. MAIN RESULTS: There were no trials comparing instruments for transabdominal CVS. Forceps and cannula were evaluated in five transcervical CVS trials involving 472 women. When a cannula was used, operators obtained an inadequate sample (less than 5 mg) more often (relative risk (RR) 4.21, 95% confidence interval (CI) 2.15 to 8.25). Compared with forceps, cannulae had to be re-inserted more often (RR 2.98, 95% CI 1.62 to 5.47). Also, inserting a cannula was more painful (RR 1.93, 95% CI 1.11 to 3.37). One study reported the cost of the procedures and found CVS with cannula to be more expensive (weighted mean difference $183.7, 95% confidence interval 152.62 to 214.78). When different types of cannulae were compared, Portex cannula was more likely to result in an inadequate sample and a difficult or painful procedure when compared with either the silver or aluminum cannula respectively. REVIEWER'S CONCLUSIONS: Although there is some evidence to support the use of small forceps for transcervical chorionic villus sampling, the evidence is not strong enough to support change in practice for clinicians who have become familiar with aspiration cannulae. PMID- 12535387 TI - Vaccines for preventing malaria. AB - BACKGROUND: Four types of malaria vaccine, SPf66 and MSP/RESA vaccines (against the asexual stages of the Plasmodium parasite) and CS-NANP and RTS,S vaccines (against the sporozoite stages), have been tested in randomized controlled trials in endemic areas. OBJECTIVES: To assess malaria vaccines against Plasmodium falciparum, P. vivax, P. malariae and P ovale in preventing infection, disease and death. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register (July 2002), the Cochrane Controlled Trials Register (The Cochrane Library Issue 2, 2002), MEDLINE (1966 to July 2002), EMBASE (1980 to May 2002), Science Citation Index (1981 to July 2002), and reference lists of articles. We also contacted organizations and researchers in the field. SELECTION CRITERIA: Randomized controlled trials comparing vaccines against Plasmodium falciparum, P. vivax, P. malariae or P. ovale with placebo or routine antimalarial control measures in people of any age receiving a challenge malaria infection. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Eighteen efficacy trials involving 10,971 participants were included. There were ten trials of SPf66 vaccine, four trials of CS-NANP vaccines, two trials of RTS,S vaccine, and two of MSP/RESA vaccine. Results with SPf66 in reducing new malaria infections (P. falciparum) were heterogeneous: it was not effective in four African trials (Peto odds ratio (OR) 0.96, 95% confidence interval (CI) 0.81 to 1.14), but in five trials outside Africa the number of first attacks was reduced (Peto OR 0.77, 95% CI 0.67 to 0.88). Trials to date have not indicated any serious adverse events with SPf66 vaccine. In three trials of CS-NANP vaccines, there was no evidence for protection by these vaccines against P. falciparum malaria (Peto OR 1.12, 95% CI 0.64 to 1.93). In a small trial in non-immune adults in the USA, RTS,S gave strong protection against experimental infection with P. falciparum. In a trial in an endemic area of the Gambia in semi-immune people, there was a reduction in clinical malaria episodes in the second year of follow up, corresponding to a vaccine efficacy of 66% (CI 14% to 85%). In a trial in Papua New Guinea, MSP/RESA had no protective effect against episodes of clinical malaria. There was evidence of an effect on parasite density, but this differed according to whether the participants had been pretreated with sulfadoxine/pyrimethamine or not. The prevalence of infections with the parasite subtype of MSP2 in the vaccine was reduced compared with the other subtype (Peto OR 0.35, CI 0.23 to 0.53). REVIEWER'S CONCLUSIONS: There is no evidence for protection by SPf66 vaccines against P. falciparum in Africa. There is a modest reduction in attacks of P. falciparum malaria following vaccination with SPf66 in other regions. Further research with SPf66 vaccines in South America or with new formulations of SPf66 may be justified. There was not enough evidence to evaluate the use of CS-NANP vaccines. The RTS,S vaccine showed promising result, as did the MSP/RESA vaccine, but it should include the other main allelic form of MSP2. The MSP/RESA trial demonstrated that chemotherapy during a vaccine trial may reduce vaccine efficacy, and trials should consider very carefully whether this practice is justified. PMID- 12535388 TI - Maternal oxygen administration for suspected impaired fetal growth. AB - BACKGROUND: Fetal hypoxaemia is often a feature of fetal growth impairment. It has been suggested that perinatal outcome after suspected impaired fetal growth might be improved by giving mothers continuous oxygen until delivery. OBJECTIVES: The objective was to assess the effects of maternal oxygen therapy in suspected impaired fetal growth on fetal growth and perinatal outcome. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register (November 2002). SELECTION CRITERIA: Acceptably controlled trials comparing maternal oxygen therapy with no oxygen therapy in suspected impaired fetal growth. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality was assessed. MAIN RESULTS: Three studies involving 94 women were included. Oxygenation compared with no oxygenation was associated with a lower perinatal mortality rate (relative risk: 0.50, 95% confidence interval 0.32 to 0.81). However, higher gestational age in the oxygenation groups may have accounted for the difference in mortality rates. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the benefits and risks of maternal oxygen therapy for suspected impaired fetal growth. Further trials of maternal hyperoxygenation seem warranted. PMID- 12535389 TI - Prophylactic methylxanthines for extubation in preterm infants. AB - BACKGROUND: When preterm infants have been given intermittent positive pressure ventilation (IPPV) for respiratory failure, weaning from support and tracheal extubation may be difficult. A significant contributing factor is thought to be the relatively poor respiratory drive and tendency to develop hypercarbia and apnea, particularly in very preterm infants. Methylxanthine treatment started before extubation might stimulate breathing and increase the chances of successful weaning from IPPV. OBJECTIVES: In preterm infants being weaned from IPPV and in whom endotracheal extubation is planned, does treatment with methylxanthine reduce the use of intubation and IPPV, without clinically important side effects? SEARCH STRATEGY: The standard search strategy of the Cochrane Neonatal Review Group was used. This included searches of the Oxford Database of Perinatal Trials, The Cochrane Central Register of Controlled Trials (The Cochrane Library, Issue 3 2002), MEDLINE (1966 to October 2002). SELECTION CRITERIA: All published trials utilising random or quasi-random patient allocation, in which treatment with methylxanthines (theophylline or caffeine) was compared with placebo or no treatment to improve the chances of successful extubation of preterm or low birth weight infants, were included. DATA COLLECTION AND ANALYSIS: The standard methods of the Cochrane Collaboration and its Neonatal Review Group were used. The second author assessed the quality of trials and extracted data independently. Results are expressed as relative risk (RR) and risk difference (RD) with 95% confidence intervals. MAIN RESULTS: Overall analysis of the six published trials shows that methylxanthine treatment results in a reduction in failure of extubation within one week [summary RR 0.47 (0.32, 0.70)]. Overall there is an absolute reduction of 27 % in the incidence of failed extubation [summary RD -.27 (-.39, -.15)]. Thus, overall in these six trials the number needed to treat (NNT) with methylxanthine to prevent one case of failed extubation is 3.7 (2.7, 6.7). There is significant heterogeneity in the RD meta analysis (p=0.007) related to the large variation in baseline rate in the control groups (range 20 - 100%). One study (Durand 1987) found that treatment was effective in reducing failed extubation in those born at less than 1000 grams and who were less than one week old. In the small prespecified subgroups in this trial, infants of less than 1 kg birth weight and older than one week and those of birth weight 1000-1250 grams who had failed extubation once, no significant benefit was found. REVIEWER'S CONCLUSIONS: Implications for practice. Methylxanthines increase the chances of successful extubation of preterm infants within one week. One trial suggests that this benefit is principally in infants of extremely low birth weight extubated in the first week. There is insufficient information to assess side effects or longer term effects on child development. Implications for research. Further trials are required comparing methylxanthines with placebo for extubation of very preterm infants. There is a need to stratify infants by gestational age (a better indicator of immaturity) rather than birth weight in future studies. Caffeine, with its wider therapeutic margin (Blanchard 1992, Steer 2002) would be the better treatment to evaluate against placebo. Side effects and neuro-developmental status at follow up should be included as outcomes. PMID- 12535390 TI - Maternal nutrient supplementation for suspected impaired fetal growth. AB - BACKGROUND: One way of attempting to improve fetal growth has been nutrient supplementation for the mother when fetal growth is impaired. Different nutrients such as carbohydrates and amino acids have been suggested as treatments for impaired fetal growth. OBJECTIVES: The objective of this review was to assess the effects of nutrient administration for suspected fetal growth impairment on fetal growth and perinatal outcome. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register. Date of last search: 1 November 2002. SELECTION CRITERIA: Acceptably controlled trials of nutrient administration for suspected impaired fetal growth compared to placebo or no treatment. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: Three studies involving 121 women were included. They were small and/or had methodological limitations. No difference was detected in the number of small for gestational age infants whose mothers had received nutrients compared to those who did not (relative risk for calf blood extract 0.54, 95% confidence interval (CI) 0.20 to 1.47; for glucose 1.11, 95% CI 0.64 to 1.92; for galactose 0.78, 95% CI 0.39 to 1.54. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of nutrient therapy for suspected impaired fetal growth. The studies were too small to detect any rare adverse effects. PMID- 12535391 TI - Drugs for preventing malaria-related illness in pregnant women and death in the newborn. AB - BACKGROUND: Malaria contributes to maternal illness and anaemia in pregnancy, especially in first-time mothers, and could harm the mother and the baby. Interventions to prevent or mitigate the effects of malaria during pregnancy are often recommended. OBJECTIVES: To assess drugs given to prevent malaria infection and its consequences in pregnant women living in malarial areas. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register (July 2002); the Cochrane Controlled Trials Register (Issue 3, 2002); MEDLINE (1966-July 2002); EMBASE (1974-July 2002); and LILACS (accessed July 2002). We contacted researchers in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials in pregnant women of drugs given regularly that aim to mitigate the effects of malaria in pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. Data extraction was done by two reviewers using standard criteria. MAIN RESULTS: 14 trials included (n=3454); only 2 were adequately concealed. For women of all parity groups, the meta-analysis (n=2890) showed lower parasitaemia and placental malaria in the intervention arm. For women having the first or second baby, there were 9 studies (n=3454). Severe antenatal anaemia was less common (RR 0.62, 95%CI 0.50 to 0.78, 4 studies), perinatal mortality appeared lower (RR 0.73, 95% CI 0.73 to 0.99, 3 studies). Maternal parasitaemia was lower with the intervention (RR 0.24, 95%CI 0.14 to 0.42, random effects model, 6 studies), and mean birthweight higher (WMD 122 g, 95%I 81 to 164 g, 8 studies), and low birthweight was less common (RR 0.49, 95%CI 0.36 to 0.65, 6 studies). REVIEWER'S CONCLUSIONS: Drugs given routinely for malaria during pregnancy reduce severe antenatal anaemia in the mother, and are associated with higher birthweight and probably reduced perinatal mortality. This effect appears to be limited to low parity women. PMID- 12535392 TI - Short-term low-dose corticosteroids vs placebo and nonsteroidal antiinflammatory drugs in rheumatoid arthritis. AB - BACKGROUND: The effect of low dose corticosteroids, equivalent to 15 mg prednisolone daily or less, in patients with rheumatoid arthritis has been questioned. We performed a systematic review of trials which compared corticosteroids with placebo or non-steroidal, anti-inflammatory drugs. OBJECTIVES: To determine whether short-term (i.e. as recorded within the first month of therapy), oral low-dose corticosteroids (corresponding to a maximum of 15 mg prednisolone daily) is superior to placebo and non-steroidal, anti inflammatory drugs in patients with rheumatoid arthritis. SEARCH STRATEGY: Medline Silverplatter, The Cochrane Controlled Trials Register, reference lists and a personal archive. Date of last search May 2002. SELECTION CRITERIA: All randomised studies comparing an oral corticosteroid (not exceeding an equivalent of 15 mg prednisolone daily) with placebo or a non-steroidal, anti-inflammatory drug were eligible if they reported clinical outcomes within one month after start of therapy. For adverse effects, long-term trials and matched cohort studies were also selected. DATA COLLECTION AND ANALYSIS: Decisions on which trials to include were made independently by two observers based on the methods sections of the trials. Standardised mean difference (random effects model) was used for the statistical analyses. MAIN RESULTS: Ten studies, involving 320 patients, were included. Prednisolone had a marked effect over placebo on joint tenderness (standardised mean difference 1.31, 95% confidence interval 0.78 to 1.83), pain (1.75, 0.87 to 2.64) and grip strength (0.41, 0.13 to 0.69). Measured in the original units, the differences were 12 tender joints (6 to 18) and 22 mm Hg (5 to 40) for grip strength. Prednisolone also had a greater effect than non steroidal, anti-inflammatory drugs on joint tenderness (0.63, 0.11 to 1.16) and pain (1.25, 0.26 to 2.24), whereas the difference in grip strength was not significant (0.31, -0.02 to 0.64). Measured in the original units, the differences were 9 tender joints (5 to 12) and 12 mm Hg (-6 to 31). The risk of adverse effects, also during moderate- and long-term use, seemed acceptable. REVIEWER'S CONCLUSIONS: Prednisolone in low doses (not exceeding 15 mg daily) may be used intermittently in patients with rheumatoid arthritis, particularly if the disease cannot be controlled by other means. Since prednisolone is highly effective, short-term placebo controlled trials studying the clinical effect of low-dose prednisolone or other oral corticosteroids are no longer necessary. PMID- 12535393 TI - Doxapram for ventilatory failure due to exacerbations of chronic obstructive pulmonary disease. AB - BACKGROUND: COPD is a progressive illness and in the later stages, exacerbations may lead to ventilatory failure. The combination of hypoxia and hypercapnia can lead to coma and death. Correction of these blood gas abnormalities is a medical emergency. Doxapram is a respiratory stimulant used to stimulate respiration in this setting. OBJECTIVES: The objective of this review was to assess the effects of doxapram on gas exchange and clinical outcomes in people with ventilatory failure due to acute exacerbations of chronic obstructive pulmonary disease. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and reference lists of articles. We also contacted experts in the field, study authors and drug companies. Following electronic searches conducted in November 2001 one further unpublished study has been included in the review. SELECTION CRITERIA: Randomised trials comparing doxapram with other treatments or placebo in people with ventilatory failure due to exacerbations of chronic obstructive pulmonary disease. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. MAIN RESULTS: Four trials involving 176 people were included. The trials were of variable quality. Doxapram was marginally superior to placebo in preventing blood gas deterioration. In the two studies comparing doxapram and non-invasive ventilation the results were conflicting: an early small study suggested non-invasive ventilation was superior. However, a subsequent larger study in severe participants suggested doxapram was equally effective in terms of blood gases changes, with no differences observed in mortality and frequent treatment failure. REVIEWER'S CONCLUSIONS: Doxapram can improve blood gas exchange over the first few hours of treatment. Newer techniques such as non-invasive ventilation may prove to be more effective, although there is no randomised trial evidence to this effect. PMID- 12535394 TI - Anticoagulants for preventing recurrence following presumed non-cardioembolic ischaemic stroke or transient ischaemic attack. AB - BACKGROUND: After a first ischaemic stroke, further vascular events due to thromboembolism (especially myocardial infarction and recurrent stroke) are common and often fatal. Anticoagulants could potentially reduce the risk of such events, but any benefits could be offset by an increased risk of fatal or disabling haemorrhages. OBJECTIVES: The objective of this review was to assess the effect of prolonged anticoagulant therapy (compared with placebo or open control) following presumed non-cardioembolic ischaemic stroke or transient ischaemic attack. SEARCH STRATEGY: We searched the Cochrane Stroke Group trials register. We contacted companies marketing anticoagulant agents. The most recent search for this review was carried out in August 2002. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing at least one month of anticoagulant therapy with control in people with previous presumed non cardioembolic ischaemic stroke or transient ischaemic attack. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials for inclusion, assessed trial quality and extracted the data. MAIN RESULTS: Eleven trials involving 2487 patients were included. The quality of the 9 trials which predated routine computerised tomography scanning and the use of the International Normalised Ratio to monitor anticoagulation was poor. There was no evidence of an effect of anticoagulant therapy on either the odds of death or dependency (two trials, odds ratio 0.83, 95% confidence interval [CI] 0.52 to 1.34) or of 'non-fatal stroke, myocardial infarction, or vascular death' (four trials, odds ratio 0.96, 95% CI 0.68-1.37). Death from any cause (odds ratio 0.95, 95% CI 0.73 to 1.24) and death from vascular causes (odds ratio 0.86, 95% CI 0.66 to 1.13) were not significantly different between treatment and control. The inclusion of two recent completed trials did not alter these conclusions. There was no evidence of an effect of anticoagulant therapy on the risk of recurrent ischaemic stroke (odds ratio 0.85, 95% CI 0.66 to 1.09). However, anticoagulants increased fatal intracranial haemorrhage (odds ratio 2.54, 95% CI 1.19 to 5.45), and major extracranial haemorrhage (odds ratio 3.43, 95% CI 1.94 to 6.08). This is equivalent to anticoagulant therapy causing about 11 additional fatal intracranial haemorrhages and 25 additional major extracranial haemorrhages per year for every 1000 patients given anticoagulant therapy. REVIEWER'S CONCLUSIONS: Compared with control, there was no evidence of benefit from long-term anticoagulant therapy in people with presumed non-cardioembolic ischaemic stroke or transient ischaemic attack, but there was a significant bleeding risk. PMID- 12535395 TI - Educational interventions for asthma in children. AB - BACKGROUND: Self-management education programs have been developed for children with asthma, but it is unclear whether such programs improve outcomes. OBJECTIVES: To determine the efficacy of asthma self-management education on health outcomes in children. SEARCH STRATEGY: Systematic search of the Cochrane Airways Group's and Cochrane Schizophrenia Group's Special Registers of Controlled Trials and hand searches of the reference lists of relevant review articles. SELECTION CRITERIA: Randomized and controlled clinical trials of asthma self-management education programs in children and adolescents aged 2 -18 years. DATA COLLECTION AND ANALYSIS: All studies were assessed independently by two reviewers. Disagreements were settled by consensus. Study authors were contacted for missing data or to verify methods. Subgroup analyses examined the impact of type and intensity of educational intervention, self-management strategy, trial type, asthma severity, adequacy of follow-up, and study quality. MAIN RESULTS: Of 45 trials identified, 32 studies involving 3706 patients were eligible. Asthma education programs were associated with moderate improvement in measures of airflow (standardized mean difference [SMD] 0.50, 95% confidence interval [CI] 0.25 to 0.75) and self-efficacy scales (SMD 0.36, 95% CI 0.15 to 0.57). Education programs were associated with modest reductions in days of school absence (SMD 0.14, 95% CI -0.23 to -0.04), days of restricted activity (SMD -0.29, 95% CI 0.49 to -0.08), and emergency room visits (SMD -0.21, 95% CI -0.33 to -0.09). There was a reduction in nights disturbed by asthma when pooled using a fixed effects but not a random-effects model. Effects of education were greater for most outcomes in moderate-severe, compared with mild-moderate asthma, and among studies employing peak flow versus symptom-based strategies. Effects were evident within the first 6 months, but for measures of morbidity and health care utilization, were more evident by 12 months. REVIEWER'S CONCLUSIONS: Asthma self management education programs in children improve a wide range of measures of outcome. Self-management education directed to prevention and management of attacks should be be incorporated into routine asthma care. Conclusions about the relative effectiveness of the various components are limited by the lack of direct comparisons. Future trials of asthma education programs should focus on morbidity and functional status outcomes, including quality of life, and involve direct comparisons of the various components of interventions. PMID- 12535396 TI - Selegiline for Alzheimer's disease. AB - BACKGROUND: Alzheimer's disease is the most common cause of dementia in older people accounting for some 60% of cases with late-onset cognitive deterioration. It is now thought that several neurotransmitter dysfunctions are involved from an early stage in the pathogenesis of Alzheimer's disease-associated cognitive decline. The efficacy of selegiline for symptoms of Alzheimer's disease remains controversial and is reflected by its low rate of prescription and the lack of approval by several regulatory authorities in Europe and elsewhere. Reasons for this uncertainty involve the modest overall effects observed in some trials, the lack of benefit observed in several trials, the use of cross-over designs which harbour methodological problems in a disease like dementia and the difficulty in interpreting results from trials when a variety of measurement scales are used to assess outcomes. OBJECTIVES: The objective of this review is to assess whether or not selegiline improves the well-being of patients with Alzheimer's disease. SEARCH STRATEGY: The Cochrane Dementia and Cognitive Impairment Group Register of Clinical Trials, was searched using the terms 'selegiline', 'l-deprenyl', "eldepryl" and "monamine oxidase inhibitor-B". MEDLINE, PsycLIT and EMBASE electronic databases were searched with the above terms in addition to using the group strategy (see group details) to limit the searches to randomised controlled trials. SELECTION CRITERIA: All unconfounded, double-blind, randomised controlled trials in which treatment with selegiline was administered for more than a day and compared to placebo in patients with dementia. DATA COLLECTION AND ANALYSIS: An individual patient data meta-analysis of selegiline, Wilcock 2002 provides much of the data that are available for this review. Seven studies provided individual patient data and this was pooled with summary statistics from the published papers of the other nine studies. Where possible, intention-to-treat data were used but usually the meta analyses were restricted to completers' data (data on people who completed the study). MAIN RESULTS: There are 17 included trials. There were very few significant treatment effects and these were all in favour of selegiline; cognition at 4-6 weeks and 8-17 weeks, and activities of daily living at 4-6 weeks. There is little evidence of adverse effects caused by selegiline, and few withdrew from trials, apart from the Sano trial. The analyses were conducted on data available. There was no attempt to correct for missing patients because there were so few and withdrawal was probably unconnected with treatment. All trials examined the cognitive effects of selegiline, and in addition 12 trials examined the behavioural and mood effects. The meta-analysis revealed benefits on memory function, shown by improvement in the memory tests from several cognitive tests (the Randt Memory Index from Agnoli 1990 and Agnoli 1992, the BSRT from Sunderland 1992, prose recall from Filip 1991, ADAS-cog from Lawlor 1997, the Wechsler Memory Scale from Loeb 1990 and Mangoni 1991, the Rey AVL from Piccinin 1990, and the MMSE from Sano 1995, Tariot 1998, Filip 1991, Freedman 1996, Burke 1993 and Riekkinen 1993). The combined memory tests, and overall the combined cognitive tests, analysed using standardised mean differences, showed an improvement due to selegiline compared with placebo at 4-6 weeks (SMD 0.39, 95%CI 0.07 to 0.72, P = 0.02, random effects model ) and 8-17 weeks, ( SMD 0.44, 95%CI 0.04 to 0.84, P = 0.03, random effects model). The meta analyses of emotional state show no treatment effects. Several studies assessed activities of daily living using several different scales, the GBS-motor function from Agnoli 1990, the NOSIE-daily living from Filip 1991, the BDS-daily living from Loeb 1990 and Mangoni 1991, the DS from Sano 1995 and PIADL from Tariot 1998. The combined tests, analysed using the standardised mean difference, showed an improvement due to selegiline at 4-6 weeks (SMD -0.27, 95% CIs -0.41 to -0.13, P = <.001). The global rating scales, the BDS used by Burke 1993 and Tariot 1998, and the GBS used by Agnoli 1990 and Agnoli 1992, and the GDS used by Freedman 1996 and the CGI by Filip 1991, analysed using standardised mean differences showed no effect of selegiline. A variety of adverse effects were recorded, but very few patients left a trial as a direct result. Four studies reported no side effects. Mangoni 1991 reported poor tolerability for 3 patients out of 68 on treatment and 1 out of 51 on placebo, resulting in dropouts. Small numbers found equally in both groups reported anxiety, agitation, dizziness, nausea and dyspepsia. Piccinin 1990 reported that selegiline was well tolerated with few adverse reactions (dizziness and orthostatic hypotension) and no resulting drop outs. Burke 1993 and Loeb 1990 both reported that selegiline was very well tolerated with no serious side effects. Sano 1995 reported 49 categories of adverse events but found no differences between the 4 arms of the factorial trial. Freedman 1996 reported unequal numbers of dropouts in the trial with 7 subjects withdrawing from the selegiline group and only 1 subject from the placebo group. The meta-analyses of the numbers suffering adverse effects, and of the numbers of withdrawals before the end of the trial show no difference between control and selegiline. REVIEWER'S CONCLUSIONS: Despite its initial promise, ie the potential neuroprotective properties, and its role in the treatment of Parkinson's disease sufferers, selegiline for Alzheimer's disease has proved disappointing. Although there is no evidence of a significant adverse event profile, there is also no evidence of a clinically meaningful benefit for Alzheimer's disease sufferers. This is true irrespective of the outcome measure evaluated, ie cognition, emotional state, activities of daily living, and global assessment, whether in the short, or longer term (up to 69 weeks), where this has been assessed. There would seem to be no justification, therefore, to use it in the treatment of people with Alzheimer's disease, nor for any further studies of its efficacy in Alzheimer's disease. PMID- 12535397 TI - Psychotherapy for bulimia nervosa and binging. AB - BACKGROUND: Bulimia nervosa and like syndromes, such as binge eating disorder, are common in young Western women. A specific manual based psychotherapy, cognitive behaviour therapy (CBT) has been developed for the treatment of bulimia nervosa (CBT-BN). Other psychotherapies, some from a different theoretical framework, and some modifications of CBT are also used. OBJECTIVES: To evaluate the efficacy of psychotherapeutic treatments for those with binge eating syndromes, that have been tested in randomised controlled trials. The efficacy of CBT in the specific treatment of bulimia nervosa and binge eating disorder was evaluated. CBT therapy was compared with waiting list or a non-treatment group, any other psychotherapy, CBT in a "pure self-help" form and CBT augmented by exposure and response therapy. In addition, the review aimed to evaluate the evidence for the efficacy of other psychotherapies when compared to a no treatment control group and to evaluate the evidence for the efficacy of other psychotherapies when compared to a 'placebo' therapy. SEARCH STRATEGY: A handsearch of The International Journal of Eating Disorders since its first issue; database searches of MEDLINE, EXTRAMED, EMBASE, PSYCHLIT, CURRENT CONTENTS, LILACS, SCISEARCH, The Cochrane Collaboration Controlled Trials Register and the Cochrane Depression, Anxiety and Neurosis Controlled Trials Register; citation list searching and personal approaches to authors were used. SELECTION CRITERIA: All studies that have tested any form of psychotherapy for adult patients with non-purging bulimia nervosa, binge eating disorder and/or EDNOS of a bulimic type, and which have applied a randomised controlled and standardized outcome methodology, were sought for the purpose of this review. DATA COLLECTION AND ANALYSIS: Data were entered into a spreadsheet programme, and into the REVMAN analysis program. Relative risk analyses were conducted of binary outcome data. The relative risk analysis was used rather than the odds ratio as the outcome measures proposed were not measuring a rare event (such as death) and the total number of studies was small. Standardized mean difference analyses were conducted of continuous variable outcome data, as the continuous outcome measures were not consistent across studies. Sensitivity analyses were conducted of a number of measures of trial quality. Data were not reported in such a way to permit subgroup analyses, but the effects of treatment on depressive symptoms, psychosocial and/or interpersonal functioning, general psychiatric symptoms and weight were examined where possible. Chi-square tests for homogeneity were done, at 5% level of significance, using a fixed effects model. Funnel plots to evaluate presence of publication bias were completed and are available in a text file upon request. MAIN RESULTS: To date, more than 1365 trials have been generated by searching and over 100 trials have been evaluated in detail. Because of a relatively high number of original exclusions (n=12) the trial inclusion criteria were broadened to include those with non-blinded outcome assessment, providing 34 trials for analyses. Because of incomplete published and available data, at best up to 12 studies had data available for any single analysis. The maximum number of total patients included in a single analysis was 602. The majority of studies evaluated patients with bulimia nervosa of a purging type. The review supported the efficacy of cognitive-behavioural psychotherapy (CBT) and particularly CBT-BN in the treatment of people with bulimia nervosa and also (but less strongly due to the small number of trials) like eating disorder syndromes. CBT had been used with efficacy in group settings. Other psychotherapies were also efficacious, particularly interpersonal psychotherapy in the longer-term. Self-help approaches that used highly structured CBT treatment manuals, were promising albeit with more modest results generally, and their evaluation in bulimia nervosa approach merits further research. Exposure and response prevention did not appear to enhance the efficacy of CBT. Psychotherapy alone is unlikely to reduce or change body weight in people with bulimia nervosa or similar eating disorders. REVIEWER'S CONCLUSIONS: There was a small body of evidence for the efficacy of cognitive-behaviour therapy in bulimia nervosa and similar syndromes, but the quality of trials was very variable (e.g. the majority were not blinded) and sample sizes were often small in comparison to pharmacotherapy trials. More trials are needed, particularly for binge eating disorder and other EDNOS syndromes, and trials evaluating other psychotherapies and less intensive psychotherapies. PMID- 12535398 TI - Vaginal misoprostol for cervical ripening and induction of labour. AB - BACKGROUND: Misoprostol (Cytotec, Searle) is a prostaglandin E1 analogue marketed for use in the prevention and treatment of peptic ulcer disease. It is inexpensive, easily stored at room temperature and has few systemic side effects. It is rapidly absorbed orally and vaginally. Although not registered for such use, misoprostol has been widely used for obstetric and gynaecological indications, such as induction of abortion and of labour. This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. OBJECTIVES: To determine the effects of vaginal misoprostol for third trimester cervical ripening or induction of labour. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register (October 2002), the Cochrane Controlled Trials Register (The Cochrane Library, Issue 3, 2002) and bibliographies of relevant papers. SELECTION CRITERIA: The criteria for inclusion included the following: (1) clinical trials comparing vaginal misoprostol used for third trimester cervical ripening or labour induction with placebo/no treatment or other methods listed above it on a predefined list of labour induction methods; (2) random allocation to the treatment or control group; (3) adequate allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusions. DATA COLLECTION AND ANALYSIS: A strategy was developed to deal with the large volume and complexity of trial data relating to labour induction. This involved a two-stage method of data extraction. The initial data extraction was done centrally, and incorporated into a series of primary reviews arranged by methods of induction of labour, following a standardised methodology. The data will be extracted from the primary reviews into a series of secondary reviews, arranged by category of woman. To avoid duplication of data in the primary reviews, the labour induction methods have been listed in a specific order, from one to 25. Each primary review includes comparisons between one of the methods (from two to 25) with only those methods above it on the list. MAIN RESULTS: Sixty-two trials have been included. Compared to placebo, misoprostol was associated with increased cervical ripening (relative risk of unfavourable or unchanged cervix after 12 to 24 hours with misoprostol 0.09, 95% confidence interval (CI) 0.03 to 0.24). It was also associated with reduced failure to achieve vaginal delivery within 24 hours (relative risk (RR) 0.36, 95% CI 0.19 to 0.68). Uterine hyperstimulation, without fetal heart rate changes, was increased (RR 11.7 95% CI 2.78 to 49). Compared with vaginal prostaglandin E2, intracervical prostaglandin E2 and oxytocin, vaginal misoprostol labour induction was associated with less epidural analgesia use, fewer failures to achieve vaginal delivery within 24 hours and more uterine hyperstimulation. Compared with vaginal or intracervical prostaglandin E2, oxytocin augmentation was less common, with misoprostol and meconium-stained liquor more common. Compared with intracervical prostaglandin E2, unchanged or unfavourable cervix after 12 to 24 hours was less common with misoprostol. Lower doses of misoprostol compared to higher doses were associated with more need for oxytocin augmentation, less uterine hyperstimulation, with and without fetal heart rate changes, and a non significant trend to fewer admissions to neonatal intensive care unit. Use of a gel preparation of misoprostol versus tablet was associated with less hyperstimulation and more use of oxytocin and epidural analgesia. Information on women's views is conspicuously lacking. REVIEWER'S CONCLUSIONS: Vaginal misoprostol appears to be more effective than conventional methods of cervical ripening and labour induction. The apparent increase in uterine hyperstimulation is of concern. Doses not exceeding 25 mcg four-hourly of concern. Doses not exceeding 25 mcg four-hourly appeared to have similar effectiveness and risk of uterine hyperstimulation to conventional labour inducing methods. The studies reviewed were not large enough to exclude the possibility of rare but serious adverse events, particularly uterine rupture, which has been reported anecdotally following misoprostol use in women with and without previous caesarean section. The authors request information on cases of uterine rupture known to readers. Further research is needed to establish the ideal route of administration and dosage, and safety. Professional and governmental bodies should agree guidelines for the use of misoprostol, based on the best available evidence and local circumstances. PMID- 12535399 TI - Self-management education and regular practitioner review for adults with asthma. AB - BACKGROUND: A key component of many asthma management guidelines is the recommendation for patient education and regular medical review. A number of controlled trials have been conducted to measure the effectiveness of asthma education programmes. These programmes improve patient knowledge, but their impact on health outcomes is less well established. This review was conducted to examine the strength of evidence supporting Step 6 of the Australian Asthma Management Plan: "Educate and Review Regularly"; to test whether health outcomes are influenced by education and self-management programmes. OBJECTIVES: The objective of this review was to assess the effects of asthma self-management programmes, when coupled with regular health practitioner review, on health outcomes in adults with asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and reference lists of articles. SELECTION CRITERIA: Randomised trials of self-management education in adults over 16 years of age with asthma. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. Study authors were contacted for confirmation. MAIN RESULTS: Thirty six trials, which compared self-management education with usual care, were included. Self-management education reduced hospitalisations (relative risk 0.64, 95% confidence interval 0.50 to 0.82); emergency room visits (relative risk 0.82, 95% confidence interval (0.73 to 0.94); unscheduled visits to the doctor (relative risk 0.68, 95% confidence interval 0.56 to 0.81); days off work or school (relative risk 0.79, 95% confidence interval 0.67 to 0.93); nocturnal asthma (relative risk 0.67, 95% confidence interval 0.0.56 to 0.79); and quality of life (standard mean difference 0.29, confidence interval 0.11 to 0.47). Measures of lung function were little changed. REVIEWER'S CONCLUSIONS: Education in asthma self-management which involves self-monitoring by either peak expiratory flow or symptoms, coupled with regular medical review and a written action plan improves health outcomes for adults with asthma. Training programmes that enable people to adjust their medication using a written action plan appear to be more effective than other forms of asthma self-management. PMID- 12535400 TI - Moderately early (7-14 days) postnatal corticosteroids for preventing chronic lung disease in preterm infants. AB - BACKGROUND: Corticosteroids have been used late in the neonatal period to treat chronic lung disease (CLD) in preterm babies, and early to try to prevent it. CLD is likely to be the result of persisting inflammation in the lung and the use of powerful anti-inflammatory drugs like dexamethasone has some rationale. Early use tends to be associated with increased adverse effects so that studies of moderately early treatment (7-14 days postnatal) might have the dual benefits of fewer side effects and onset of action before chronic inflammation is established. OBJECTIVES: To determine if moderately early (7-14 days) postnatal corticosteroid treatment vs control (placebo or nothing) is of benefit in the prevention and/or treatment of early chronic lung disease in the preterm infant. SEARCH STRATEGY: Randomised controlled trials of postnatal corticosteroid therapy were sought from the Oxford Database of Perinatal Trials, Cochrane Database of Controlled Trials, MEDLINE (1966 - October 2002), hand searching paediatric and perinatal journals, examining previous review articles and information received from practicing neonatologists. Authors of all studies were contacted, where possible, to confirm details of reported follow-up studies, or to obtain any information about long-term follow-up where none had been reported. SELECTION CRITERIA: Randomised controlled trials of postnatal corticosteroid treatment from 7-14 days of birth in high risk preterm infants were selected for this review. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including mortality, CLD (including late rescue with corticosteroids, or need for home oxygen therapy), death or CLD, failure to extubate, complications during the primary hospitalisation (including infection, hyperglycaemia, hypertension, hypertrophic cardiomyopathy, pneumothorax, severe intraventricular haemorrhage (IVH), necrotizing enterocolitis (NEC), gastrointestinal bleeding, and severe retinopathy of prematurity (ROP)), and long term outcome (including blindness, deafness, cerebral palsy and major neurosensory disability), were abstracted and analysed using RevMan 4.1. MAIN RESULTS: Seven studies enrolling a total of 669 participants were eligible for inclusion in this review. Moderately early steroid treatment (vs placebo or nothing) reduced mortality by 28 days, chronic lung disease at 28 days and 36 weeks, and death or chronic lung disease at 28 days or 36 weeks. Earlier extubation was facilitated. There was no significant effect on the rates of pneumothorax, severe ROP, or NEC. Adverse effects included hypertension, hyperglycaemia, gastrointestinal bleeding, hypertrophic cardiomyopathy and infection. Steroid-treated infants were less likely to need late rescue with dexamethasone. There were limited data from four studies of long term follow-up; these did not show evidence of an increase in adverse neurological outcomes. REVIEWER'S CONCLUSIONS: Moderately early corticosteroid therapy (started at 7-14 days) reduces neonatal mortality and CLD, but at the cost of important short term adverse effects. Limited evidence concerning long term effects is provided by the trials included in this review. The methodological quality of the studies determining the long-term outcome is limited in some cases, the children have been assessed predominantly before school age, and no study has been sufficiently powered to detect important adverse long-term neurosensory outcomes. Therefore, given the risk:benefit ratio of short-term effects and the limited long-term follow-up data, it seems appropriate to reserve moderately early corticosteroid treatment to infants who cannot be weaned from mechanical ventilation and to minimise the dose and duration of any course of therapy. More research is urgently needed, including long term follow-up of survivors included in previous and any future trials, before the benefits and risks of postnatal steroid treatment, including initiation at 7-14 days, can be reliably assessed (See DART study; Doyle 2000a). PMID- 12535401 TI - Delayed (>3 weeks) postnatal corticosteroids for chronic lung disease in preterm infants. AB - BACKGROUND: Many preterm babies who survive, having had respiratory distress syndrome (RDS) or not, go on to develop chronic lung disease (CLD). This is probably due to persistence of inflammation in the lung. Corticosteroids have powerful anti-inflammatory effects and have been used to treat established CLD. However it is unclear whether any beneficial effects outweigh the adverse effects of these drugs. OBJECTIVES: To determine if late (usually > 3 weeks) postnatal corticosteroid treatment vs control (placebo or nothing) is of benefit in the treatment of chronic lung disease (CLD) in the preterm infant. SEARCH STRATEGY: Randomised controlled trials of postnatal corticosteroid therapy were sought from the Oxford Database of Perinatal Trials, the Cochrane Controlled Trials Register, MEDLINE 1966 through October 2002, hand searching paediatric and perinatal journals, examining previous review articles and information received from practising neonatologists. Authors of all studies were contacted, where possible, to confirm details of reported follow-up studies, or to obtain any information about long-term follow-up where none had been reported. SELECTION CRITERIA: Randomised controlled trials of postnatal corticosteroid treatment initiated at predominantly > 3 weeks of age in preterm infants with CLD were selected for this review. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including mortality, CLD (including need for home oxygen, or need for late rescue with corticosteroids), death or CLD, failure to extubate, complications in the primary hospitalisation (including infection, hyperglycaemia, glycosuria, hypertension, echodensities on ultrasound scan of brain, necrotising enterocolitis (NEC), gastrointestinal bleeding, intestinal perforation, and severe retinopathy of prematurity (ROP)), and long term outcome (including blindness, deafness, cerebral palsy and major neurosensory disability), were abstracted and analysed using RevMan 4.1. MAIN RESULTS: Nine trials enrolling a total of 562 participants were eligible for this review. Delayed steroid treatment had no significant effect on mortality. Beneficial effects of delayed steroid treatment included reductions in failure to extubate by 7 or 28 days, chronic lung disease at 36 weeks, need for late rescue treatment with dexamethasone, discharge to home on oxygen therapy, and death or CLD at 36 wk. There was no evidence of increase in risk of infection, necrotising enterocolitis, or gastrointestinal bleeding. Short term adverse affects included glycosuria and hypertension. There was an increase in severe retinopathy of prematurity, of borderline significance, but no significant increase in blindness. The trend to an increase in cerebral palsy was partly offset by a trend in the opposite direction in death before late follow up. The combined rate of death or cerebral palsy was not significantly different between steroid and control groups. Major neurosensory disability, and the combined rate of death or major neurosensory disability, were not significantly different between steroid and control groups. REVIEWER'S CONCLUSIONS: The benefits of late corticosteroid therapy may not outweigh actual or potential adverse effects. Although there continues to be concern about an increased incidence of adverse neurological outcomes in infants treated with postnatal steroids (see also review of Early postnatal corticosteroids), this review of postnatal corticosteroid treatment for CLD initiated predominantly after three weeks of age suggests that late or delayed therapy may not significantly increase the risk of adverse long-term neurodevelopmental outcomes. However, the methodological quality of the studies determining the long-term outcome is limited in some cases, the children have been assessed predominantly before school age, and no study has been sufficiently powered to detect important adverse long-term neurosensory outcomes. Given the evidence of both benefits and harms of treatment, and the limitations of the evidence at present, it appears prudent to reserve the use of late corticosteroids to infants who cannot be weaned from mechanical ventilation, and to minimise the dose and duration of any course of treatment. PMID- 12535402 TI - Early postnatal (<96 hours) corticosteroids for preventing chronic lung disease in preterm infants. AB - BACKGROUND: Chronic lung disease (CLD) remains a major problem in neonatal intensive care units. Persistent inflammation in the lungs is the most likely underlying pathogenesis. Corticosteroids have been used to either prevent or treat CLD because of their potent anti-inflammatory effects. OBJECTIVES: To determine if postnatal corticosteroid treatment is of benefit in the prevention of chronic lung disease (CLD) in the preterm infant. This review examines the outcome of trials where preterm infants at risk of CLD were given postnatal steroids within 96 hours after birth. SEARCH STRATEGY: Randomised controlled trials of postnatal corticosteroid therapy were sought from the Oxford Database of Perinatal Trials, the Cochrane Controlled Trials Register, MEDLINE (1966 - October 2002), hand searching paediatric and perinatal journals, examining previous review articles and information received from practising neonatologists. Authors of all studies were contacted, where possible, to confirm details of reported follow-up studies, or to obtain any information about long-term follow up where none had been reported. SELECTION CRITERIA: Randomised controlled trials of postnatal corticosteroid treatment within 96 hours of birth (early) in high risk preterm infants were selected for this review. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including mortality, CLD (including late rescue with corticosteroids, and need for home oxygen therapy), death or CLD, failure to extubate, complications during the primary hospitalisation (including infection, hyperglycaemia, hypertension, pulmonary air leak, patent ductus arteriosus (PDA), severe intraventricular haemorrhage (IVH), periventricular leucomalacia (PVL), necrotising enterocolitis (NEC), gastrointestinal bleeding, intestinal perforation, severe retinopathy of prematurity (ROP), and long-term outcome (including blindness, deafness, cerebral palsy and major neurosensory disability) were abstracted and analysed using RevMan 4.1. MAIN RESULTS: Twenty-one randomised controlled trials enrolling a total of 3072 participants were eligible for inclusion in this review. A meta-analysis of these trials demonstrated significant benefits as regards earlier extubation and decreased risks of CLD at both 28 days and 36 weeks, death or CLD at 28 days and 36 weeks, PDA and severe ROP. There were no significant differences in the rates of neonatal or subsequent mortality, infection, severe IVH, PVL, NEC or pulmonary haemorrhage. Gastrointestinal bleeding and intestinal perforation were important adverse effects and the risks of hyperglycaemia and hypertension were also increased. In the nine trials which have reported late outcomes, several adverse neurological effects were found at follow-up examinations of survivors treated with early steroids: developmental delay (not defined), cerebral palsy and abnormal neurological exam. However, major neurosensory disability was not significantly increased, either overall in the 4 studies where this outcome could be determined, or in the 2 individual studies where the rate of cerebral palsy and abnormal neurological exam were significantly increased. Moreover, the rate of the combined outcome of death or major neurosensory disability was not significantly increased. REVIEWER'S CONCLUSIONS: The benefits of early postnatal corticosteroid treatment (< 96 hours) may not outweigh the known or potential adverse effects of this treatment. Although early steroid treatment facilitates extubation and reduces the risk of chronic lung disease, it causes short-term adverse effects including gastrointestinal bleeding, intestinal perforation, hyperglycaemia, hypertension, hypertrophic cardiomyopathy and growth failure. Long-term follow-up studies report an increased risk of abnormal neurological exam and cerebral palsy. However, the methodological quality of the studies determining the long-term outcome is limited in some cases, the children have been assessed predominantly before school age, and no study has been sufficiently powered to detect important adverse long-term neurosensory outcomes. There is a compelling need for the long term follow-up and reporting of late outcomes, especially neurological and developmental outcomes, among surviving infants who participated in all randomised trials of early postnatal corticosteroid treatment. The role of inhaled steroids remains to be elucidated. PMID- 12535403 TI - Mycobacterium vaccae immunotherapy for treating tuberculosis. AB - BACKGROUND: Some authorities have advocated Mycobacterium vaccae immunotherapy for treating tuberculosis and other infections caused by mycobacteria. OBJECTIVES: To assess the effects of Mycobacterium vaccae as an adjunct to chemotherapy for treating tuberculosis. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register (September 2002), the Cochrane Controlled Trials register (Issue 3, 2002), MEDLINE (1966 to October 2002), EMBASE (1980 to September 2002), and reference lists of articles. We also contacted organisations and individuals working in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials using whole, killed Mycobacterium vaccae for patients with tuberculosis. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. MAIN RESULTS: Seven trials met the inclusion criteria. There was no effect on mortality (4 trials, OR 1.09, 95% CI 0.79 to 1.49). No consistent effect on sputum negativity or sputum culture was shown. Most immunotherapy recipients experienced local adverse reactions (2 trials, OR 18.2, 95% CI 9 to 37), some of which progressed to ulceration and scarring. REVIEWER'S CONCLUSIONS: Mycobacterium vaccae does not benefit patients with tuberculosis. PMID- 12535404 TI - Topical agents or dressings for pain in venous leg ulcers. AB - BACKGROUND: Venous leg ulcers affect up to 1 per cent of people at some time in their life. These ulcers are often painful and some clinicians choose dressings and topical treatments (analgesia/ local anaesthetic) to reduce the pain both during and between dressing changes. OBJECTIVES: To assess the effectiveness of dressings, local anaesthetics or topical analgesia for pain relief in venous leg ulceration. SEARCH STRATEGY: Cochrane Wounds Group Register and the Cochrane Collaboration Field in Complementary Medicine were searched in June 2002. Cochrane Pain Palliative and Supportive Care Group and Cochrane Wounds Group strategy were combined and used. SELECTION CRITERIA: All randomised controlled trials which evaluated local interventions used to relieve venous leg ulcer pain were considered. Pain was defined as either persistent pain or pain at dressing changes or debridement. DATA COLLECTION AND ANALYSIS: Eligibility for inclusion was confirmed by two reviewers who independently assessed the potential trials. Details of eligible studies were summarised using a data extraction sheet which was checked by the second reviewer. MAIN RESULTS: No trials evaluating interventions for persistent pain were identified for the initial review in 1999 nor the update in 2002. Three trials were included in the 1999 review comparing a eutectic mixture of local anaesthetic (EMLA) versus placebo for pain at debridement. In 2002 a further 3 trials were available (6 trials in total with 317 patients). The studies were considered sufficiently similar to pool and meta analysis found a statistically significant reduction in debridement pain scores with EMLA 5% cream. EMLA was associated with a reduction in pain scores (measured on a 100 mm scale) of 20.6 mm (95% Confidence Interval 29.11-12.19). One small trial measured healing as an outcome and found no difference in numbers of ulcers healed at the end of the study. REVIEWER'S CONCLUSIONS: EMLA provides effective pain relief for venous leg ulcer debridement however, the effect of the product on ulcer healing is unknown. Research is required to determine the impact of debridement and of EMLA on ulcer healing. There were no trials addressing the treatment of persistent pain (between and at dressing changes) and further research is warranted. PMID- 12535405 TI - Interventions to modify sexual risk behaviors for preventing HIV infection in men who have sex with men. AB - BACKGROUND: MSM remain at great risk for HIV infection. Program planners and policy makers need descriptions of interventions and quantitative estimates of intervention effects to make informed decisions concerning prevention efforts. OBJECTIVES: 1. To locate and describe outcome studies evaluating the effects of behavioral and social interventions targeting MSM. 2. To summarize the effectiveness of these interventions among MSM. 3. To stratify results by characteristics of interventions and participants. 4. To identify gaps and indicate future research, policy, and practice needs. SEARCH STRATEGY: We searched electronic databases (MedLine, PsycInfo, etc.); several current journals (e.g., AIDS, AIDS and Behavior, AIDS Education and Prevention, American Journal of Public Health, Journal of Acquired Immune Deficiency Syndromes, etc.); manuscripts submitted by researchers; bibliographies of relevant articles; and other published reviews, for published and unpublished reports from 1988 through 1997. SELECTION CRITERIA: Studies were considered in scope if they examined the effects of behavioral interventions to reduce risk for HIV or STD transmission. We reviewed studies in scope for criteria of outcome relevance (measurement of at least one of a list of behavioral or biologic outcomes, e.g., unprotected sex or incidence of HIV infections) and methodologic rigor (randomized controlled trials or certain strong quasi-experimental designs with comparison groups). DATA COLLECTION AND ANALYSIS: As of June 1998 we had identified 13 eligible studies. Twelve studies (7 trials of small group interventions, 3 community-level interventions, and 2 individual level interventions) reported intervention effects on unprotected sex. Because few studies reported effects on condom use (3 studies), number of sex partners without regard to condom use (4 studies), or HIV or STD incidence (1 study in which no infections occurred) we do not address these outcomes at this time. We present those analyses which can be performed in the current Cochrane RevMan software, followed by more complete analyses that permit inclusion of community-level studies, adjustment for baseline conditions, calculation of effect sizes from a wider variety of statistics (e.g., an F statistic from a one-way ANOVA), and simultaneous meta-analysis of continuous and dichotomous outcomes (Johnson 2002b). We translate the summary effect to reduction in risk behavior based on the background prevalence of unprotected sex. Finally we provide analyses stratified by intervention content (interpersonal skills addressed or not), intervention format (community vs small group or individual) and mean age of participants (23 to 31 vs. 32 to 36). MAIN RESULTS: A summary measure of intervention effects on reducing unprotected sex was favorable (odds ratio = 0.73) and statistically significant (CI, 0.60 to 0.88), corresponding to a 23% reduction in the proportion of men engaging in unprotected sex. Effects were homogeneous among studies, but were slightly more favorable among community-level interventions, those that served populations in their 20s rather than their 30s, and those that promoted interpersonal skills. REVIEWER'S CONCLUSIONS: These studies demonstrate that interventions can promote risk reduction among MSM. Yet given the epidemiology of HIV in Pattern I countries, the small number of rigorous controlled intervention trials for this population is striking. Many more rigorous evaluations of HIV prevention efforts with MSM are needed to ascertain with confidence the effects of specific intervention components, population characteristics, and methodologic features. PMID- 12535406 TI - Community interventions for preventing smoking in young people. AB - BACKGROUND: Decisions to smoke are made within a broad social context. Community interventions use co-ordinated, widespread, multi-component programmes to try and influence behaviour. OBJECTIVES: To determine the effectiveness of community interventions in preventing the uptake of smoking in young people. SEARCH STRATEGY: The Tobacco Addiction group specialised register, Medline and other health, psychology and public policy electronic databases were searched, the bibliographies of identified studies were checked and contact was made with content area specialists. Searches were updated in September 2002. SELECTION CRITERIA: Randomised and non randomised controlled trials that assessed the effectiveness of multi-component community interventions compared to no intervention or to single component or school-based programmes only. Reported outcomes had to include smoking behaviour in young people under the age of 25 years. DATA COLLECTION AND ANALYSIS: Information relating to the characteristics and the content of community interventions, participants, outcomes and methods of the study was extracted by one reviewer and checked by a second. Studies were combined using qualitative narrative synthesis. MAIN RESULTS: Seventeen studies were included in the review, 46 studies did not meet all of the inclusion criteria. All studies used a controlled trial design, with six using random allocation of schools or communities. Of thirteen studies which compared community interventions to no intervention controls, two, which were part of cardiovascular disease prevention programmes, reported lower smoking prevalence. Of three studies comparing community interventions to school-based programmes only, one found differences in reported smoking prevalence. One study reported a lower rate of increase in prevalence in a community receiving a multi-component intervention compared to a community exposed to a mass media campaign alone. One study reported a significant difference in smoking prevalence between a group receiving a media, school and homework intervention compared to a group receiving the media component only. REVIEWER'S CONCLUSIONS: There is some limited support for the effectiveness of community interventions in helping prevent the uptake of smoking in young people. PMID- 12535407 TI - Colloid solutions for fluid resuscitation. AB - BACKGROUND: Colloids are widely used in the replacement of fluid volume. However doubts remain as to which colloid is best. Different colloids vary in their molecular weight and therefore in the length of time they remain in the circulatory system. Because of this and their other characteristics, they may differ in their safety and efficacy. OBJECTIVES: To compare the effects of different colloid solutions in patients thought to need volume replacement. SEARCH STRATEGY: We searched the Cochrane Injuries Group specialised register, the Cochrane Controlled Trials Register (2002 Issue 3), MEDLINE (1994-2002/07), EMBASE (1974-2002 August week 1), and the National Research Register (2002 issue 3). Bibliographies of trials retrieved were searched, and drug companies manufacturing colloids were contacted for information. The search was last updated in September 2002. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing colloid solutions in critically ill and surgical patients thought to need volume replacement. The main outcomes measured were death, amount of whole blood transfused, and incidence of adverse reactions. DATA COLLECTION AND ANALYSIS: Two authors independently extracted the data and assessed the quality of the trials. MAIN RESULTS: Fifty-seven trials met the inclusion criteria, with a total of 3659 participants. Quality of allocation concealment was judged to be adequate in 20 trials and poor or uncertain in 37. Deaths were obtained from 36 trials. For albumin or PPF versus hydroxyethyl starch (HES) 20 trials (n=1029) reported mortality. The pooled relative risk (RR) was 1.17 (95% CI 0.91, 1.50). For albumin or PPF versus gelatin four trials (n=542) reported mortality. The RR was 0.99 (0.69, 1.42). For gelatin vs HES 11 trials (n=945) reported mortality, RR was 1.00 (0.78,1.28). RR was not estimable in the albumin vs dextran, gelatin vs dextran, and HES vs dextran groups. Thirty-six trials recorded the amount of blood transfused, however quantitative analysis was not possible due to skewness and variable reporting. Fifteen trials recorded adverse reactions, but none occurred. REVIEWER'S CONCLUSIONS: From this review, there is no evidence that one colloid solution is more effective or safe than any other, although the confidence intervals are wide and do not exclude clinically significant differences between colloids. Larger trials of fluid therapy are needed if clinically significant differences in mortality are to be detected or excluded. PMID- 12535408 TI - Olanzapine for schizophrenia. AB - BACKGROUND: Olanzapine is an atypical antipsychotic that is reported to be effective without producing the disabling extrapyramidal side effects associated with the older, typical antipsychotic drugs. OBJECTIVES: To determine the clinical effects and safety of olanzapine as compared with placebo, typical and other atypical antipsychotic drugs for schizophrenia and schizophreniform psychoses. SEARCH STRATEGY: The reviewers undertook electronic searches of Biological Abstracts (1980-1999), The Cochrane Library (Issue 2, 1999), EMBASE (1980-1999), MEDLINE (1966-1999), PsycLIT (1974-1999) and The Cochrane Schizophrenia Group's Register (October 2000). References of all identified studies were searched for further trials, and the reviewers contacted relevant pharmaceutical companies and authors of trials. SELECTION CRITERIA: All randomised clinical trials comparing olanzapine to placebo or any antipsychotic treatment for those with schizophrenia or schizophreniform psychoses. DATA COLLECTION AND ANALYSIS: Data were independently extracted. For homogeneous dichotomous data the random effects relative risk (RR), the 95% confidence intervals (CI) and, where appropriate, the number needed to treat (NNT) were calculated on an intention-to-treat basis. For continuous data the reviewers calculated weighted mean differences. MAIN RESULTS: Twenty one trials are included. Attrition from olanzapine versus placebo studies was so great (olanzapine - 61%, placebo - 73% by six weeks, RR 0.85 CI 0.7-0.98, NNT 8 CI 5 40) that interpretation of results is problematic. Olanzapine appeared superior to placebo at six weeks for the outcome of 'no important clinical response' (RR 0.88 CI 0.8-0.98, NNT 8 CI 5-27) and global mental state scores. Although dizziness and dry mouth were reported more frequently in the olanzapine-treated group, this did not reach statistical significance. Tthe olanzapine group gained more weight. When compared to typical antipsychotic drugs, data from several small trials are incomplete; but, for the short term outcome of 'no important clinical response', olanzapine seem as effective as typical antipsychotics (n=2778, RR 0.9 CI 0.76-1.06). Brief Psychiatric Rating Scale (BPRS) data tended to be equivocal but Positive and Negative Syndrome Scale (PANSS) rating of total score and negative and positive symptom sub-scores favoured olanzapine. With high attrition in both groups (olanzapine - 36%, typical drug - 49% by 6 weeks, n=2738, RR 0.85 CI 0.66-1.1; olanzapine - 83%, typical drug - 90% by 1 year, n=2738, RR 0.9 CI 0.86-1.02), the assumptions included in all continuous data are considerable. Participants allocated olanzapine experienced fewer extrapyramidal side effects than people given haloperidol. Weight change data for the short term are not conclusive (n=2455, WMD 0.8kg CI -0.6-2.2) but the three to 12 month results suggest an average gain of four kilograms (n=233, WMD 4 CI 0.3-7.8). It is difficult to distinguish between olanzapine and other atypical drugs, although it may cause fewer extrapyramidal side effects than risperidone (n=339, RR 0.6 CI 0.4-0.9, NNH 8 CI 4-29). Olanzapine did cause more weight gain than its comparators but current data are not statistically significant (3-12 months, n=535, WMD 2.2kg CI -0.6-5). One study (n=180) found no clear differences between olanzapine and clozapine for people with treatment-resistant illness. REVIEWER'S CONCLUSIONS: The large proportions of participants leaving the studies early, in the large multi-centre trials makes it difficult to draw firm conclusions on clinical effects. For people with schizophrenia olanzapine may offer antipsychotic efficacy with fewer extrapyramidal side effects than typical drugs but more weight gain. Large, long-term randomised trials with participants, interventions and primary outcomes that are familiar to those wishing to help those with schizophrenia are long overdue. PMID- 12535409 TI - Nebulised hypertonic saline for cystic fibrosis. AB - BACKGROUND: The lung disease in cystic fibrosis is characterised by impaired mucociliary clearance. Hypertonic saline (HS) has been shown to enhance mucociliary clearance in-vitro and this may act to lessen the destructive inflammatory process in the airways. OBJECTIVES: To investigate the effects of treatment with nebulised hypertonic saline on people with CF compared to placebo and or other treatments that enhance mucociliary clearance. SEARCH STRATEGY: 'We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group trials register which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings. Date of the most recent search of the Group's register: October 2001. SELECTION CRITERIA: All controlled trials (any language) assessing the effect of hypertonic saline compared to placebo or other mucolytic therapy, for any duration or dose regimen in people with cystic fibrosis of any age or severity. DATA COLLECTION AND ANALYSIS: All identified trials were independently reviewed by both reviewers & all data collected. Trial quality was assessed along with allocation concealment. MAIN RESULTS: Fourteen controlled trials were identified. Nine trials met the inclusion criteria; these involved 235 participants with an age range of 6 to 46 years. Two short-term trials of immediate effect on mucociliary clearance demonstrated that HS increased isotope clearance compared to control. Lung function as measured by improvement in Forced Expiratory Volume at one second (FEV1 l/min) was observed in four trials. When 3% to 7% saline was used in a volume of 10mls twice a day, in comparison to placebo, HS led to a significant increase in FEV1, WMD 12.20 (95%CI 4.30 to 20.10). In comparison to deoxyribonuclease (DNase) two trials used a similar concentration and volume of HS. Over a three week period the groups showed a similar increase in FEV1, WMD -1.60 (95%CI -11.16 to 7.96). However after 12 weeks treatment in participants with moderate to severe lung disease compared to DNase, HS 5mls twice a day showed less benefit to FEV1, WMD -13.00 (95%CI -22.46 to -3.54). No serious adverse events were noted. REVIEWER'S CONCLUSIONS: Nebulised hypertonic saline improves mucociliary clearance in short term clinical trials and appears to increase lung function compared to control. In comparison to DNase it may be less effective at improving lung function, after three months. At this stage there is insufficient evidence to support the use of hypertonic saline as routine treatment for people with cystic fibrosis. PMID- 12535410 TI - Corticosteroid therapy for nephrotic syndrome in children. AB - BACKGROUND: In nephrotic syndrome protein leaks from the blood to the urine through the glomeruli resulting in hypoproteinaemia and generalised oedema. Children with untreated nephrotic syndrome frequently die from infections. The majority of children with nephrotic syndrome respond to corticosteroids. However about 70% of children experience a relapsing course with recurrent episodes of oedema and proteinuria. Corticosteroid usage has reduced the mortality rate in childhood nephrotic syndrome to around 3%, with infection remaining the most important cause of death. However corticosteroids have known adverse effects such as obesity, poor growth, hypertension, diabetes mellitus, osteoporosis and adrenal suppression. The original treatment schedules for childhood nephrotic syndrome were developed in an ad hoc manner. The optimal doses and durations of corticosteroid therapy that are most beneficial and least harmful have not been clarified. OBJECTIVES: The aim of this review was to determine the benefits and harms of different corticosteroid regimens in preventing relapse in children with steroid sensitive nephrotic syndrome (SSNS). SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (Cochrane Library, Issue 2, 2002), Cochrane Renal Group Specialised Register (July 2002), MEDLINE (1966 - July 2002) and EMBASE (1980 - July 2002) without language restriction, reference lists of articles, abstracts from proceedings and contact with known investigators in the area. SELECTION CRITERIA: Randomised controlled trials were included if they were carried out in children (aged three months to 18 years) in their initial or subsequent episode of SSNS, if they compared different durations, total doses or other dose strategies using prednisone or other corticosteroid agent and if they had outcome data at six months or more. DATA COLLECTION AND ANALYSIS: Two reviewers independently reviewed all eligible studies for inclusion, assessed study quality and extracted data. The principle outcome measure was the number of children with and without relapse after six and 12-24 months. Secondary outcomes sought included the number of children who developed frequently relapsing nephrotic syndrome and adverse events. A random effects model was used to estimate summary effect measures (relative risk (RR), risk difference (RD)) after testing for heterogeneity. Meta-regression was used to explore potential between study differences due to the baseline risk of relapse, study quality and types of interventions used. MAIN RESULTS: Five additional trials were included in the review for a total of 17 trials. A meta-analysis of six trials, which compared two months of prednisone with three months or more in the first episode, showed that the longer duration significantly reduced the risk of relapse at 12 - 24 months (RR 0.70; 95% CI 0.58 to 0.84) without an increase in adverse events. There was an inverse linear relationship between the duration of treatment and risk of relapse (RR = 1.26 - 0.112 duration; r(2) = 0.56; p = 0.03). The number of children who became frequent relapsers and the mean relapse rate/patient/year were also significantly reduced without increase in serious adverse events. In children with frequently relapsing nephrotic syndrome, deflazacort was significantly more effective in maintaining remission than prednisone (RR 0.44; 95% CI 0.25 to 0.78). REVIEWER'S CONCLUSIONS: Children in their first episode of SSNS should be treated for at least three months with an increase in benefit being demonstrated for up to seven months of treatment. In a population with a baseline risk for relapse following the first episode of 60% with two months of prednisone, daily prednisone for four weeks followed by alternate day therapy for six months would be expected to reduce the number of children experiencing a relapse by about 33%. In children who relapse frequently, deflazacort deserves further study. PMID- 12535411 TI - Mobilisation strategies after hip fracture surgery in adults. AB - BACKGROUND: Post-operative care programmes after hip fracture surgery include strategies for mobilisation, such as early weight bearing, gait retraining and other physical therapy interventions. OBJECTIVES: To evaluate the effects of different mobilisation strategies and programmes after hip fracture surgery. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group specialised register (to August 2002), MEDLINE (1966 to August week 5 2002), the National Research Register (Issue 2, 2002), Current Controlled Trials, conference proceedings and reference lists of articles. SELECTION CRITERIA: All randomised or quasi-randomised trials comparing different mobilisation strategies/programmes after hip fracture surgery. DATA COLLECTION AND ANALYSIS: The reviewers independently assessed trial quality, using a 10 item scale, and extracted data. Wherever appropriate and possible, the data are presented graphically. MAIN RESULTS: Our second update included one new study. Only two of the six included trials involved a similar comparison. All trials had methodological limitations, including inadequate follow-up. Two trials, involving a total of 188 patients, compared a more intensive with a less intensive regimen of physiotherapy. One reported a lack of demonstrable difference in recovery of the two patient groups at nine weeks follow-up. The other found a higher level of drop-out in the more intensive group with no difference in length of hospital stay. Only limited outcome data were available for both trials. One trial of 80 patients evaluated a quadriceps muscle strengthening exercise programme. Improved mobility, leg extension power and Barthel score were reported for the intervention group. A treadmill gait retraining programme was compared with a conventional gait retraining programme in 40 patients. More patients in the treadmill group had recovered their pre-fracture level of mobility by the time of hospital discharge, which tended to happen earlier than for the control group. Neither of these differences were statistically significant. One trial of 27 patients compared neuromuscular stimulation of the quadriceps muscle with placebo stimulation. More patients in the stimulation group had recovered their pre-fracture mobility at 13 weeks follow-up. One trial involving 273 patients with a displaced intracapsular fracture treated by internal fixation compared weight bearing at two weeks after surgery with delayed weight bearing at 12 weeks after surgery. From the limited data available, there were no statistically significant differences between the two methods of treatment for non-union, mortality and overall unfavourable outcome at one year. REVIEWER'S CONCLUSIONS: There is insufficient evidence from randomised trials to determine the effects of more frequent or a more intensive programme of physiotherapy, quadriceps strengthening exercises, treadmill gait retraining, or neuromuscular stimulation after hip fracture surgery. There is also insufficient evidence to determine the effects of early weight bearing after the internal fixation of an intracapsular proximal femoral fracture. PMID- 12535412 TI - Interventions for vaginismus. AB - BACKGROUND: Vaginismus is involuntary vaginal muscle contraction which makes sexual intercourse difficult or impossible. It is one of the more common female psychosexual problems. Various therapeutic strategies for vaginismus, such as sex therapy and desensitization, have been considered in uncontrolled studies. OBJECTIVES: The aim of this review is to determine the clinical effectiveness of treatments for vaginismus and also to examine the role of partner participation in the effectiveness of the treatment. SEARCH STRATEGY: The Cochrane Collaboration Depression, Anxiety & Neurosis Controlled Trials Register (Issue 3, 2000), the Cochrane Controlled Trials Register (Cochrane Library, Issue 2, 2002), MEDLINE (1966 to Nov 2002), EMBASE (1980 to Nov 2002), PsycINFO (1967 to Nov 2002) and CINAHL (1982 to Nov 2002) were searched. The Journal of Sex Research (1974 to 1999), Sexual & Marital Therapy (1986 to 1999), Sexual Dysfunction (1998 to 1999) and the Journal of Sex and Marital Therapy (1974 to 1999) were handsearched. Reference lists and conference abstracts were searched. Experts in the field were contacted regarding unpublished material. SELECTION CRITERIA: Controlled trials comparing treatments for vaginismus with another treatment, a placebo treatment, treatment as usual or waiting list control. DATA COLLECTION AND ANALYSIS: The reviewers extracted data which were verified with the trial investigator. MAIN RESULTS: Two controlled trials were identified although data were only available from one trial. The second trial compared two forms of systematic desensitization and reported no discernible differences between them. REVIEWER'S CONCLUSIONS: In spite of encouraging results reported from uncontrolled series of patients there is very limited evidence from controlled trials concerning the effectiveness treatments for vaginismus. Further trials are need to compare therapies with waiting list control and with other therapies. PMID- 12535413 TI - Laparoscopic techniques versus open techniques for inguinal hernia repair. AB - BACKGROUND: Inguinal hernia repair is the most frequently performed operation in general surgery. The standard method for inguinal hernia repair had changed little over a hundred years until the introduction of synthetic mesh. This mesh can be placed by either using an open approach or by using a minimal access laparoscopic technique. Although many studies have explored the relative merits and potential risks of laparoscopic surgery for the repair of inguinal hernia, most individual trials have been too small to show clear benefits of one type of surgical repair over another. OBJECTIVES: The objective of this review was to compare minimal access laparoscopic mesh techniques with open techniques. Comparisons of open mesh techniques versus open non-mesh techniques have been considered in a separate Cochrane review. SEARCH STRATEGY: We searched MEDLINE, EMBASE, and The Cochrane Central Controlled Trials Registry for relevant randomised controlled trials. The reference list of identified trials, journal supplements, relevant book chapters and conference proceedings were searched for further relevant trials. Through the EU Hernia Trialists Collaboration (EUHTC) communication took place with authors of identified randomised controlled trials to ask for information on any other recent and ongoing trials known to them. Specialists involved in research on the repair of inguinal hernia were contacted to ask for information about any further completed and ongoing trials. The world wide web was also searched. SELECTION CRITERIA: All published and unpublished randomised controlled trials and quasi-randomised controlled trials comparing laparoscopic groin hernia repair with open groin hernia repair were eligible for inclusion. Trials were included irrespective of the language in which they were reported. DATA COLLECTION AND ANALYSIS: Individual patient data were obtained, where possible, from the responsible trialist for all eligible studies. All reanalyses were cross-checked by the reviewers and verified by the trialists before inclusion. Where IPD were unavailable additional aggregate data were sought from trialists and published aggregate data checked and verified by the trialists. IPD were available for 25 trials, additional aggregated data for seven and published data only for nine. Where possible, time to event analysis for hernia recurrence and return to usual activities were performed on an intention to treat principle. The main analyses were based on all trials. Sensitivity analyses based on the data source and trial quality were also performed. Pre defined subgroup analyses based on recurrent hernias, bilateral hernias and femoral hernias were also carried out. MAIN RESULTS: 41 published reports of eligible trials were included involving 7161 participants. Sample sizes ranged from 38 to 994, with follow-up from 6 weeks to 36 months. Duration of operation was longer in the laparoscopic groups (WMD 14.81 minutes, 95% CI 13.98 to 15.64; p<0001). Operative complications were uncommon for both methods but more frequent in the laparoscopic group for visceral (Overall 8/2315 versus 1/2599) and vascular (Overall 7/2498 versus 5/2758) injuries. Length of hospital stay did not differ between groups (WMD -0.04 days, 95% CI -0.08 to 0.00; p=0.05, but return to usual activity was earlier for laparoscopic groups (HR 0.56, 95%CI 0.51 to 0.61; p<0.0001 - equivalent to 7 days). The data available showed less persisting pain (Overall 290/2101 versus 459/2399; Peto OR 0.54, 95% CI 0.46 to 0.64; p<0.0001), and less persisting numbness (Overall 102/1419 versus 217/1624; Peto OR 0.38, 95% CI 0.4286 to 0.49; p<0.0001) in the laparoscopic groups. In total, 86 recurrences were reported amongst 3138 allocated laparoscopic repair and 109 amongst 3504 allocated to open repair (Peto OR 0.81, 95% CI 0.61 to 1.08; p = 0.16). The use of mesh during laparoscopic hernia repair is associated with a reduction in the risk of hernia recurrence, significantly so for the transabdominal preperitoneal repair (TAPP) versus open non-mesh repair (overall 26/1440 versus preperitoneal repair (TAPP) versus open non-mesh repair (overall 26/1440 versus 47/1119; Peto OR 0.45, 95% CI 0.28 to 0.72; p=0.0009). However, no difference was detected when comparing laparoscopic methods with open mesh methods of hernia repair. REVIEWER'S CONCLUSIONS: The use of mesh during laparoscopic hernia repair is associated with a relative reduction in the risk of hernia recurrence of around 30-50%. However, there is no apparent difference in recurrence between laparoscopic and open mesh methods of hernia repair. The data suggests less persisting pain and numbness following laparoscopic repair. Return to usual activities is faster. However, operation times are longer and there appears to be a higher risk of serious complication rate in respect of visceral (especially bladder) and vascular injuries. PMID- 12535414 TI - Continuous nasogastric milk feeding versus intermittent bolus milk feeding for premature infants less than 1500 grams. AB - BACKGROUND: Most premature infants less than 1500 grams birth weight must be fed initially by tube because of their inablity to suck effectively, or to coordinate sucking, swallowing and breathing. Milk feedings can be given by tube either intermittently, typically over 10-20 minutes every two or three hours, or continuously, using an infusion pump. Although theoretical benefits and risks of each method have been proposed, effects on clinically important outcomes remain uncertain. OBJECTIVES: To examine the evidence from randomized trials regarding the effectiveness of continuous versus intermittent bolus nasogastric milk feeding in premature infants less than 1500 grams. The primary outcomes reviewed included feeding tolerance, days to reach full enteral feeding, somatic growth, days to discharge and incidence of necrotizing enterocolitis (NEC). SEARCH STRATEGY: Searches were performed of MEDLINE, CINAHL, and HealthSTAR, each up to February 2002, and the Cochrane Controlled Trials Register (The Cochrane Library, Issue 1, 2002). As well, studies identified from abstracts and conference proceedings and references from relevant publications were retrieved. SELECTION CRITERIA: Randomized and quasi-randomized clinical trials that met the following criteria for relevance: a) Enrollment of infants < 1500 grams birth weight with no major congenital anomalies which might interfere with feeding tolerance b) Comparison of continuous nasogastric versus intermittent bolus tube feedings using breastmilk or formula c) Assessment of relevant outcomes including feeding tolerance, days to full feeds, somatic growth, days to discharge, and complications such as NEC or apnea DATA COLLECTION AND ANALYSIS: All articles retrieved from the complete search were assessed independently by the two reviewers for relevance (see selection criteria), and for methodologic quality using the following criteria: blinding of randomization, blinding of intervention, complete follow-up and blinding of outcome measurement. Only those articles judged by both reviewers to be relevant and to have appropriate methodologic quality were included in the analysis. Differences were resolved through discussion and consensus of the reviewers. MAIN RESULTS: Infants fed by continuous tube feeding method took longer to reach full enteral feeds (weighted mean difference 3.0 days; 95% CI 0.7, 5.2). Although there was no evidence of a difference in the days to discharge overall, one study suggested a trend toward earlier discharge for infants less than 1000 grams birth weight fed by the continuous tube feeding method (mean difference (MD) -11 days; 95% CI -21.8, 0.2). Overall, there was no evidence of a difference in somatic growth (weight, length, head circumference or skinfold thickness) between the two groups, but subgroup analyses in one study suggested that infants less than 1000 grams and 1000 - 1250 grams birthweight gained weight faster when fed by the continuous tube feeding method (MD 2.0 g/day; 95% CI 0.5, 3.5; MD 2.0 g/day; 95% CI 0.2, 3.8, respectively). There was no evidence of a difference in the incidence of NEC. One study showed a trend toward more apneas during the study period in infants fed by the continuous tube feeding method (MD 14.0 apneas during study period; 95% CI -0.2, 28.2). REVIEWER'S CONCLUSIONS: Infants fed by the continuous tube feeding method took longer to reach full feeds, but there was no significant difference in somatic growth, days to discharge, or the incidence of NEC for infants fed by continuous versus intermittent bolus tube feeds. Small sample sizes, methodologic limitations and conflicting results of the studies to date, together with inconsistencies in controlling variables that may affect outcomes, make it difficult to make universal recommendations regarding the best tube feeding method for premature infants less than 1500 grams. The clinical benefits and risks of continuous versus intermittent nasogastric tube milk feeding cannot be reliably discerned from the limited information available from randomized trials to date. PMID- 12535415 TI - Dipyridamole for preventing stroke and other vascular events in patients with vascular disease. AB - BACKGROUND: Patients with transient ischaemic attacks (TIA) and minor ischaemic strokes are at risk of serious vascular events (death from all vascular causes, non-fatal stroke, or non-fatal myocardial infarction). Their risk of vascular events lies between 4 and 11 percent per year. Aspirin only, in a daily dose of 30 mg or more, offers only modest protection in such patients: it reduces the incidence of major vascular events by 13 percent. In a single trial, adding dipyridamole (an alternative antiplatelet agent) to aspirin was associated with a 22 percent reduction in the risk of major vascular events compared with aspirin alone. However, a systematic review of all trials of antiplatelet agents by the Antithrombotic Trialists' Collaboration showed that, in high risk patients, there was virtually no difference between the aspirin-dipyridamole combination and aspirin alone. We therefore sought to assess the effects of dipyridamole in more detail. OBJECTIVES: 1) To assess the efficacy of dipyridamole versus control in the secondary prevention of vascular events in patients with vascular disease in the presence and absence of other antiplatelet drugs. 2) To assess the safety of dipyridamole versus control in the secondary prevention of vascular events in patients with vascular disease in the presence and absence of other antiplatelet drugs. SEARCH STRATEGY: The Cochrane Stroke Group Trials Register was searched and other relevant Cochrane Groups were contacted to search their specialised registers (last search 29 April 2002). In addition the Cochrane Controlled Trials Register, MEDLINE and EMBASE were searched and Dutch manufacturers of dipyridamole were contacted to identify further published and unpublished studies. SELECTION CRITERIA: Randomised long term secondary prevention trials with concealed treatment allocation, treatment for > 1 month, starting within 6 months after presentation of a arterial vascular disease were selected (coronary artery disease, myocardial infarction, angina pectoris, retinopathy, nephropathy, peripheral arterial disease, stroke, TIA, amaurosis fugax). Therapy consisted of dipyridamole in any dose in the presence or absence of other antiplatelet drugs compared with no drug or an antiplatelet drug(s) other than dipyridamole (control group). DATA COLLECTION AND ANALYSIS: Two reviewers selected trials which met the inclusion criteria and extracted details of randomisation methods, blinding of treatments and assessments, whether intention-to-treat analysis was possible from the published data, whether treatment groups were comparable with regard to major prognostic factors, the number of patients who were excluded or lost to follow up, definition of outcome events, and entry and exclusion criteria. The methodological quality of each trial was assessed by the two reviewers on the basis of these extracted data. In addition, dose and type of antiplatelet treatments, duration of follow-up and the numbers of defined outcome events were recorded. Data published by the Antithrombotic Trialists' Collaboration were used. The remaining data were independently extracted by the same two reviewers and cross checked. Data were analysed according to the intention-to-treat principle. Relative and absolute risk reductions were calculated by means of the statistical software provided by the Cochrane Collaboration (RevMan). MAIN RESULTS: Twenty-six trials were included, with a total of 19842 patients, among whom 1399 vascular deaths and 3085 fatal and non-fatal vascular events occurred during follow-up. Compared with control, dipyridamole had no clear effect on vascular death (RR 1.02, 95% CI 0.90 to 1.17). This result was not influenced by the dose of dipyridamole or type of presenting vascular disease. Compared with control, dipyridamole appeared to reduce the risk of vascular events (RR 0.90, 95% CI 0.83 to 0.98), but this effect was only statistically significant due to a single large trial in patients presenting with cerebral ischaemia (6602 patients). Comparing dipyridamole plus aspirin with aspirin alone: there was no clear difference in vascular death, the RR was 1.03 (95% CI 0.87 to 1.22); the combination was associated with fewer vascular events with an RR of 0.90 (95% CI 0.80 to 1.00). Combination treatment of dipyridamole and aspirin compared with placebo had an RR of 0.89 (95% CI 0.79 to 1.01) for vascular death and an RR of 0.74 (95% CI 0.68 to 0.80) for vascular events. REVIEWER'S CONCLUSIONS: This review found that, for patients who presented with arterial vascular disease, there was no evidence that dipyridamole, in the presence or absence of an other antiplatelet drug (chiefly aspirin) reduced the risk of vascular death, though it may reduce the risk of further vascular events. However, this benefit was found in only a single large trial and only in patients presenting after cerebral ischaemia. There was no evidence that dipyridamole alone was more efficacious than aspirin. Further trials comparing the effects of the combination of dipyridamole with aspirin with aspirin alone are justified. PMID- 12535416 TI - Work conditioning, work hardening and functional restoration for workers with back and neck pain. AB - BACKGROUND: Physical conditioning programs, variously called work conditioning, work hardening and functional restoration/exercise programs, aim to improve work status and function. Previous attempts have been made to evaluate the efficacy of work-oriented back pain management programs, but none have focused exclusively on work or functional outcomes. OBJECTIVES: To compare the effectiveness of physical conditioning programs with management strategies that do not include physical conditioning programs, for workers with back and neck pain, in reducing time lost from work and increasing functional status. SEARCH STRATEGY: We searched the following databases to 31 May 2000: MEDLINE from 1966, EMBASE from 1980, CINAHL from 1982, Biomedical Collection: I (from 1993), II (from 1995), III (from 1995), IV (from 1995), PsycINFO from 1967, the Cochrane Central Register of Controlled Trials, PEDro. SELECTION CRITERIA: Randomized controlled trials (RCTs) that studied adults with work disability related to back or neck pain, who were included in physical conditioning programs. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed trial quality. Where data could be pooled, a meta-analysis was performed using RevMan. For continuous outcomes, we nominated a saving of 10 sick days as the smallest treatment effect that would be clinically worthwhile. For dichotomous outcomes, we considered that an intervention that affected less than one in 10 people would not be clinically worthwhile. MAIN RESULTS: Eighteen RCTs were identified in 20 publications. Twenty-three relevant contrasts were investigated. There is evidence that physical conditioning programs that include a cognitive-behavioural approach can reduce the number of sick days lost at 12 months follow-up by an average of 45 days, when compared to general practitioner usual care or advice, for workers with chronic back pain. For work-related outcomes, there is little evidence for or against the efficacy of specific exercises that are not accompanied by a cognitive-behavioural approach, in reducing sick days lost due to back pain, for workers with either acute or chronic back pain. REVIEWER'S CONCLUSIONS: Physical conditioning programs that include a cognitive-behavioural approach plus intensive physical training (specific to the job or not) that includes aerobic capacity, muscle strength and endurance, and coordination; are in some way work related; and are given and supervised by a physiotherapist or a multidisciplinary team, seem to be effective in reducing the number of sick days for some workers with chronic back pain, when compared to usual care. However, there is no evidence of their efficacy for acute back pain. PMID- 12535417 TI - Dressings and topical agents for arterial leg ulcers. AB - BACKGROUND: It is estimated that people in industrialised countries have a 1% chance of suffering from a leg ulcer at some time in their life. The majority of leg ulcers are associated with circulation problems; poor blood return in the veins causes venous ulcers (around 70% of ulcers) and poor blood supply to the legs causes arterial ulcers (around 25% of ulcers). Treatment of arterial leg ulcers is directed towards correcting the poor arterial blood supply, for example, by surgically correcting arterial blockages, and by supporting ulcer healing using topical agents (medicines in cream/ointment) and wound dressings. There are a large number of topical agents and wound dressings available and it is unclear what impact these have on ulcer healing. OBJECTIVES: To determine whether topical agents and wound dressings affect the rate of healing in arterial ulcers. To compare healing rates, costs and patient-centred outcomes between wound dressings and topical agents. SEARCH STRATEGY: Publications describing (or potentially describing) randomised controlled trials (RCTs), or controlled clinical trials (CCTs) of dressings and topical agents for arterial leg ulcers were sought through the Specialised Trials Registers of the Cochrane Wounds Group (last searched January 2002) and the Cochrane Peripheral Vascular Diseases Group (last searched April 2002). SELECTION CRITERIA: RCTs or CCTs (trials with non randomised concurrent comparison groups) were eligible for inclusion. The participants had to have ulcers that were described as arterial, and the time to healing, proportion completely healed, or rate of reduction in ulcer area had to be reported. All wound dressings and topical agents were eligible for inclusion in this review. DATA COLLECTION AND ANALYSIS: Information on the participants' characteristics, the interventions, and outcomes, as well as data on the trial methods, such as blinding of patients and clinicians, and allocation concealment were extracted using a standardised data extraction form. MAIN RESULTS: One trial met the inclusion criteria. This small trial compared ketanserin ointment with vehicle alone, changed twice a day. The trial was too small and for too short a follow-up period to be able to determine whether there was any difference in healing rates. REVIEWER'S CONCLUSIONS: There is insufficient evidence to determine whether the choice of topical agent or dressing affects the healing of arterial leg ulcers. Inadequate description of the people in the one included trial means that the results cannot be easily applied to other clinical populations. PMID- 12535418 TI - Interventions for smoking cessation in hospitalised patients. AB - BACKGROUND: An admission to hospital provides an opportunity to help people stop smoking. Individuals may be more open to help at a time of perceived vulnerability, and may find it easier to quit in an environment where smoking is restricted or prohibited. Providing smoking cessation services during hospitalisation may help more people to attempt and sustain a quit attempt. OBJECTIVES: To determine the effectiveness of interventions for smoking cessation in hospitalised patients. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group register, CINAHL and the Smoking and Health database in March 2002 for studies of interventions for smoking cessation in hospitalised patients, using terms including (hospital and patient*) or hospitali* or inpatient* or admission* or admitted. SELECTION CRITERIA: Randomised and quasi-randomised trials of behavioural, pharmacological or multicomponent interventions to help patients stop smoking conducted with hospitalised patients who were current smokers or recent quitters. We excluded studies of patients admitted for psychiatric disorders or substance abuse, those that did not report abstinence rates and those with follow-up of less than six months. DATA COLLECTION AND ANALYSIS: Two authors extracted data independently for each paper, with disagreements resolved by consensus. MAIN RESULTS: Seventeen trials met the inclusion criteria. Intensive intervention (inpatient contact plus follow-up for at least one month) was associated with a significantly higher quit rate compared to control (Peto Odds Ratio 1.82, 95% CI 1.49-2.22, six trials). Interventions with less than a month of follow-up did not show evidence of significant benefit (Peto Odds Ratio 1.09, 95% CI 0.91-1.31, seven trials). There was no evidence to judge the effect of very brief (<20 minutes) interventions delivered only during the hospital stay. Longer interventions delivered only during the hospital stay were not significantly associated with a higher quit rate (Peto Odds Ratio 1.07, 95% CI 0.79-1.44, three trials). Although the interventions increased quit rates irrespective of whether nicotine replacement therapy (NRT) was used, the results for NRT were compatible with other data indicating that it increases quit rates. There was no strong evidence that clinical diagnosis affected the likelihood of quitting. REVIEWER'S CONCLUSIONS: High intensity behavioural interventions that include at least one month of follow-up contact are effective in promoting smoking cessation in hospitalised patients. The findings of the review were compatible with research in other settings showing that NRT increases quit rates. PMID- 12535419 TI - Personalised risk communication for informed decision making about entering screening programs. AB - BACKGROUND: There is a trend towards greater patient involvement in health care decisions. Adequate discussion of the risks and benefits associated with different choices is often required if involvement is to be genuine and effective. Achieving adequate involvement of consumers and informed decision making are now seen as important goals for any screening programme. Individualised risk estimates have been shown to be effective methods of risk communication in general, but the effectiveness of different strategies has not previously been examined. OBJECTIVES: To assess the effects of different types of individualised risk communication for consumers making decisions about participating in screening. SEARCH STRATEGY: We searched the Cochrane Consumers and Communication Review Group specialised register (searched March 2001), MEDLINE (1985 to 2001), EMBASE (1985 to 2001), CancerLit (1985 to 2001), CINAHL (1985 to 2001), ClinPSYC (1989 to 2001), and the Science Citation Index Expanded (searched March 2002). Follow-up searches involved hand searching Preventive Medicine, citation searches on seven authors, and searching reference lists of articles. SELECTION CRITERIA: Randomised controlled trials addressing the decision by consumers of whether or not to undergo screening, incorporating an intervention with a 'personalised risk communication element' and reporting cognitive, affective, or behavioural outcomes. A 'personalised risk communication element' is based on the individual's own risk factors for a condition (such as age or family history). It may be calculated from an individual's risk factors using formulae derived from epidemiological data, and presented as an absolute risk or as a risk score, or it may be categorised into, for example, high, medium or low risk groups. It may be less detailed still, involving a listing, for example, of a consumer's risk factors as a focus for discussion and intervention. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. Data about the nature and setting of the intervention, and the relevant outcome data were extracted, along with items relating to methodological quality. MAIN RESULTS: Thirteen studies were included. Personalised risk communication (whether written, spoken or visually presented) was associated with increased uptake of screening tests (odds ratio (OR) 1.5 (95% confidence interval (CI) 1.11 to 2.03). There was no evidence from these studies that this increase in uptake of tests was related to informed decision making by consumers. More detailed personalised risk communication was associated with a smaller increase in uptake of tests. That is, for personalised risk communication which used and presented numerical calculations of risk, the OR for test uptake was 1.22 (95% CI 0.56 to 2.68). For risk estimates or calculations which were categorised into high, medium or low strata of risk, the OR was 1.42 (95% CI 1.07 to 1.88). For risk communication that simply listed risk personal risk factors the OR was 1.7 (95% CI 1.17 to 2.48). Most of the included studies addressed mammography programmes. These studies showed slightly smaller effects than the overall dataset, again with numerical calculated risk estimates being associated with lower ORs for uptake of tests (OR 1.13; 95% CI 0.98 to 1.29) than the other categories of (less detailed) personalised risk communication. The four studies examining risk communication in high risk individuals showed larger odds ratios for uptake of tests than the other studies. The OR for numerical calculated risk estimates was 1.48 (95% CI 1.06 to 2.07), compared to 4.66 (95% CI 2.24 to 9.69) for categorised risk estimates and 2.64 (95% CI 1.42 to 4.9) for listed personal risk factors. There were insufficient data from the included studies to report odds ratios on other key outcomes such as: intention to take tests, anxiety, satisfaction with decisions, decisional conflict, knowledge and risk perception. REVIEWER'S CONCLUSIONS: Personalised risk communication (as currently implemented in the included studies) is associated with increased uptake of screening programmes, but this may not be interpretable as evidence of informed decision making by consumers. PMID- 12535420 TI - Carbamazepine versus phenobarbitone monotherapy for epilepsy. AB - BACKGROUND: In developing countries, phenobarbitone is commonly used but its use in Europe and the USA has decreased due to concerns over adverse effects. Carbamazepine is recommended as the drug of choice for partial onset seizures, and there is concern that it may worsen some generalized onset seizure types. We report a review using individual patient data in which carbamazepine and phenobarbitone are compared. OBJECTIVES: To review the effects of carbamazepine compared to phenobarbitone monotherapy for people with partial onset seizures or generalized onset tonic-clonic seizures. SEARCH STRATEGY: The Cochrane Controlled trials register (Cochrane Library Issue 2, 2002); MEDLINE; EMBASE; handsearching; contacting experts and original trial investigators; contacting manufacturers of carbamazepine. SELECTION CRITERIA: Randomized or quasi-randomized, blinded or unblinded controlled trials in children or adults with partial onset seizures or generalized onset tonic-clonic seizures. DATA COLLECTION AND ANALYSIS: Outcome measures were (i) time to withdrawal of allocated treatment, (ii) time to 12 month remission, and (iii) time to first seizure. Data were analysed using a stratified logrank analysis with results expressed as hazard ratios (HR) and 95% confidence intervals (CIs), where a HR>1 indicates an event is more likely on phenobarbitone. A test for interaction between treatment and seizure type (partial versus generalized onset) was also undertaken. MAIN RESULTS: Data are available for 684 participants from four trials, representing 59% of the participants recruited into the nine trials that met our inclusion criteria. The main overall results (HR 95% CI) adjusted for seizure type were, (i) time to withdrawal 1.63(1.23 to 2.15), (ii) time to 12 month remission 0.87(0.65 to 1.17), (iii) time to first seizure 0.85(0.68 to 1.05). The review suggests that time to withdrawal is significantly improved with carbamazepine compared to phenobarbitone. No overall difference between drugs is identified for the outcomes 'time to 12 month remission' and 'time to first seizure'. Statistical heterogeneity was not encountered. An interaction between treatment and seizure type, confirmed statistically, was identified for time to first seizure, where phenobarbitone was favoured for partial onset seizures and carbamazepine for generalized onset tonic-clonic seizures. REVIEWER'S CONCLUSIONS: We found no overall difference between carbamazepine and phenobarbitone for time to 12 month remission or time to first seizure, however, subgroup analyses for time to first seizure suggest an advantage with phenobarbitone for partial onset seizures and a clinical advantage with carbamazepine for generalized onset tonic-clonic seizures. Phenobarbitone is significantly more likely to be withdrawn, indicating that it is less well tolerated than carbamazepine. PMID- 12535421 TI - Pharmacological interventions for non-ulcer dyspepsia. AB - BACKGROUND: The commonest cause of upper gastrointestinal symptoms is non-ulcer dyspepsia (NUD) and yet the pathophysiology of this condition has been poorly characterised and the optimum treatment is uncertain. It is estimated that pound 450 million is spent on dyspepsia drugs in the UK each year. OBJECTIVES: This review aims to determine the effectiveness of six classes of drugs (antacids, histamine H2 antagonists, proton pump inhibitors, prokinetics, mucosal protecting agents and antimuscarinics) in the improvement of either the individual or global dyspepsia symptom scores and also quality of life scores patients with non-ulcer dyspepsia. SEARCH STRATEGY: Trials were located through electronic searches of the Cochrane Controlled Trials Register (CCTR), MEDLINE, EMBASE, CINAHL and SIGLE, using appropriate subject headings and text words, searching bibliographies of retrieved articles, and through contacts with experts in the fields of dyspepsia and pharmaceutical companies. SELECTION CRITERIA: All randomised controlled trials (RCTs) comparing drugs of any of the six groups with each other or with placebo for non-ulcer dyspepsia (NUD). DATA COLLECTION AND ANALYSIS: Data were collected on dyspeptic symptom scores either individual or global symptom assessments and also quality of life scores and adverse effects. MAIN RESULTS: A total of 11796 citations were obtained. 155 trials were retrieved and 96 trials fulfilled our eligibility criteria. However, subsequent data extraction was not possible in 31 trials. The final 65 trials were included in the meta-analysis. Prokinetics (14 trials with dichotomous outcomes generating 1053 patients; relative risk reduction [RRR] = 48%; 95% confidence intervals [CI] = 27% to 63%), H2RAs (11 trials generating 2,164 patients; RRR = 22%; 95% CI = 7% to 35%) and PPIs (7 trials generating 3,031 patients; RRR = 14%; 95% CI = 5% to 23%) were significantly more effective than placebo. Bismuth salts (6 trials generating 311 patients; RRR = 40%; 95% CI = -3 to 65%) were superior to placebo but this was of marginal statistical significance. Antacids (one trial generating 109 patients; RRR = -2%; 95% CI = -36% to 24%) and sucralfate (two trials generating 246 patients; RRR = 29%; 95% CI = -40% to 64%) were not statistically significantly superior to placebo. A funnel plot suggested that the prokinetic and H2RA results could be due to publication bias. REVIEWER'S CONCLUSIONS: There is evidence that anti-secretory therapy may be effective in NUD. The trials evaluating prokinetic therapy are difficult to interpret as the meta-analysis result could have been due to publication bias. The effect of these drugs is likely to be small and many patients will need to take them on a long-term basis so economic analyses would be helpful and ideally the therapies assessed need to be inexpensive and well tolerated. PMID- 12535422 TI - Diaphragm versus diaphragm with spermicides for contraception. AB - BACKGROUND: The diaphragm is usually used with a spermicide. However, some practitioners have suggested that spermicides offer no additional contraceptive protection and have advocated alternative guidelines for the use of diaphragms. OBJECTIVES: The objective of this review was to compare the effectiveness, safety and acceptability of the diaphragm with and without spermicide. SEARCH STRATEGY: We searched MEDLINE, EMBASE, POPLINE, CINAHL, the Cochrane Controlled Trials Register, and reference lists of relevant articles. In addition, we contacted experts in the field to identify unpublished studies. SELECTION CRITERIA: Randomized controlled trials comparing women of reproductive age using the diaphragm with and without spermicide as the sole contraceptive method that reported clinical outcomes. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data on outcomes and trial characteristics and any discrepancies were resolved by consensus or by consultation with the third reviewer. The results of the one identified study are presented descriptively. MAIN RESULTS: We identified only one study. No significant difference was found in the pregnancy rates (with typical use or consistent use) or discontinuation rates between the diaphragm-with-spermicide and diaphragm-without-spermicide groups. There was a trend towards higher pregnancy rates in the diaphragm-without spermicide group. However, this study failed to recruit the planned number of participants and was consequently underpowered. REVIEWER'S CONCLUSIONS: As only one underpowered study was identified, we cannot distinguish between the contraceptive effectiveness of the diaphragm with and without spermicide. We cannot draw any conclusion at this point, further research is needed. However, the study provides no evidence to change the commonly recommended practice of using the diaphragm with spermicide. PMID- 12535423 TI - Leflunomide for treating rheumatoid arthritis. AB - BACKGROUND: Rheumatoid arthritis (RA) is a chronic inflammatory joint disease. Leflunomide, as an inhibitor of pyrimidine synthesis, has a different mechanism of action than other existing disease modifying anti-rheumatic drugs (DMARD). OBJECTIVES: To determine the efficacy and toxicity of leflunomide compared to placebo or other DMARDs in the treatment of RA. SEARCH STRATEGY: We conducted a search in MEDLINE, EMBASE, Current Contents and the Cochrane Controlled Trial Register for trials up to December 2001. We also hand-searched reference lists and consulted content experts. SELECTION CRITERIA: Two independent reviewers selected the trials that met predetermined inclusion criteria. DATA COLLECTION AND ANALYSIS: Two independent reviewers extracted data and assessed methodologic quality using standardized forms. MAIN RESULTS: Six trials were included in this review. Using the ACR20 improvement criteria, there was an absolute difference in improvement of 28% (95% confidence interval: 21 - 35%) favouring leflunomide (232 out of 413 leflunomide treated patients compared to 89 out of 311 placebo patients met the criteria). There was no difference in ACR20 response rate between the patients treated with leflunomide and SSZ or MTX at 6 and 12 months. Other clinical outcomes were improved significantly in the leflunomide group compared to placebo but not different from SSZ or MTX. Withdrawals due to adverse events with leflunomide were 10% greater than placebo (70 out of 416 compared to 18 out of 311 respectively). Important adverse events included gastrointestinal symptoms, elevated liver function tests, alopecia, and infections. Overall adverse events and withdrawals in the leflunomide group were not significantly different from SSZ or MTX. REVIEWER'S CONCLUSIONS: Leflunomide appears to improve all clinical outcomes and delay radiologic progression at both 6 and 12 months of treatment compared to placebo. Its efficacy and adverse events at 2 years of treatment are comparable to SSZ and MTX. Long-term efficacy and toxicity remains to be established. PMID- 12535424 TI - Intravenous midazolam infusion for sedation of infants in the neonatal intensive care unit. AB - BACKGROUND: The need for sedation for neonates undergoing uncomfortable procedures in the neonatal intensive care unit (NICU) has often been overlooked. Proper sedation may reduce stress and avoid complications during procedures such as mechanical ventilation. Midazolam is a short acting benzodiazepine that has been increasingly used in the NICU. However, the effectiveness of intravenous midazolam as a sedative in neonates has not been systematically evaluated. OBJECTIVES: To determine whether intravenous midazolam infusion is an effective sedative, as evaluated by behavioural and/or physiologic measurements, for critically ill neonates undergoing intensive care, and to assess clinically significant short and long term adverse effects associated with its use. SEARCH STRATEGY: Literature search according to the Cochrane Neonatal Review Group search strategy. Randomized and quasi-randomized controlled trials of intravenous midazolam use in neonates were identified by searching the Cochrane Controlled Trials Register (The Cochrane Library, Issue 3, 2002), MEDLINE (1985-2002), EMBASE (1980-2002), reference lists of published studies, personal files, and abstracts published in Pediatric Research from 1990-2002. SELECTION CRITERIA: Randomized and quasi-randomized controlled trials of intravenous midazolam infusion in infants 17 years with refractory Crohn's disease defined by conventional clinical, radiological and endoscopic criteria, which was categorized as being active (Crohn's disease activity index >150). DATA COLLECTION AND ANALYSIS: OUTCOME MEASURES: The outcome measure was the rate of induction of remission and complete withdrawal from steroids in the treatment and control groups after 16 weeks of treatment. A secondary outcome was induction of remission with reduction in steroid dose of at least 50%. Selection of trials: The results of the searches above were reviewed independently by two observers and relevant studies selected according to the predefined selection criteria. Any disagreement among reviewers was resolved by consensus. The same two reviewers assessed the methodological quality of each trial (details of randomization method, including whether intention-to-treat analysis was possible from the published data, number of patients lost to follow-up, and if a blinded outcome assessment was used). A standard data extraction form was used. Appropriateness of combining results: Trials were first reviewed to assess the clinical comparability of trial protocols and study populations. MAIN RESULTS: Three randomized placebo-controlled trials were identified. The three studies differed with respect to participants, intervention, and outcomes to the extent that it was considered to be inappropriate to combine the data statistically. Two studies which employed low doses of methotrexate orally showed no statistically significant difference between methotrexate and placebo treated patients, and one which employed a higher dose intramuscularly showed substantial benefit (number needed to treat, NNT=5). Adverse effects were more common with high dose intramuscular methotrexate therapy than with placebo. REVIEWER'S CONCLUSIONS: There is evidence from a single large randomized trial on which to recommend the use of methotrexate 25 mg intramuscularly weekly for induction of remission and complete withdrawal from steroids in patients with refractory Crohn's disease. Although adverse effects are more common than with placebo, they were not severe. There is no evidence on which to base a recommendation for use of lower dose oral methotrexate. PMID- 12535476 TI - Amantadine in Parkinson's disease. AB - BACKGROUND: Although levodopa is the most common drug prescribed to relieve the symptoms of Parkinson's disease it is associated with motor and psychiatric side effects. Consequently, interest has turned to alternative drugs with improved side-effect profiles to replace or augment levodopa. Amantadine, originally used as an antiviral drug, has been shown to improve the symptoms of Parkinson's disease. OBJECTIVES: To compare the efficacy and safety of amantadine therapy (monotherapy or adjuvant therapy) versus placebo in treating people with Parkinson's disease. SEARCH STRATEGY: Electronic searches of The Cochrane Controlled Trials Register (The Cochrane Library Issue 3, 2001), MEDLINE (1966 2001), EMBASE (1974-2001), SCISEARCH (1974-2001), BIOSIS (1993-2001), GEROLIT (1979-2001), OLDMEDLINE (1957-1965), LILACS (1982-2001), MedCarib (17th Century - 2001), PASCAL (1973-2001), JICST-EPLUS (1985-2001), RUSSMED (1973-2001), DISSERTATION ABSTRACTS (2000-2001), SIGLE (1980-2001), ISI-ISTP (1990-2001), Aslib Index to Theses (2001), Clinicaltrials.gov (2001), metaRegister of Controlled Trials (2001), NIDRR (2001) and NRR (2001) were conducted. Grey literature was hand searched and the reference lists of identified studies and reviews examined. The manufacturers of amantadine were contacted. SELECTION CRITERIA: Randomised controlled trials comparing amantadine with placebo in the treatment of patients with a clinical diagnosis of idiopathic Parkinson's disease. DATA COLLECTION AND ANALYSIS: Data was abstracted independently by NC and KD onto standardised forms and disagreements were resolved by discussion. MAIN RESULTS: Six randomised controlled trials were found comparing amantadine monotherapy or adjuvant therapy with placebo in the treatment of idiopathic Parkinson's disease. Five examined amantadine as adjuvant therapy with optimal levels of levodopa or anticholinergics and one examined amantadine as an adjuvant therapy with minimum tolerated levels of anticholinergics or as a monotherapy. Five were double-blind cross-over studies and one was a double-blind parallel group study. In total they examined 215 patients. The parallel group study allowed the randomisation codes to be broken and allowed patients in the placebo group to then receive amantadine. This could have led to bias. One study did not present the results of the placebo arm of the trial, hence we could not determine the difference between the two treatment groups. Two cross-over studies presented the results of the combined data from both treatment and placebo arms. The risk of carry-over effect into the second arm meant that these results could not be analysed. The final two studies presented at least some of their data from the end of the first arm of the trials. However only means were given, without standard deviations, so we could not determine the statistical significance of any difference between the amantadine and placebo groups. Although the authors did report on the side-effects from amantadine (such as livido recticularis, dry mouth and blurred vision), they state that none of them were severe. REVIEWER'S CONCLUSIONS: A considerable amount of evidence on the effectiveness of amantadine has accrued from non-controlled trials, often in patients with Parkinsonian conditions other than idiopathic Parkinson's disease. However, rigorous analysis of the six randomised controlled trials of amantadine reveals insufficient evidence of its efficacy and safety in the treatment of idiopathic Parkinson's disease. PMID- 12535477 TI - Parent-mediated early intervention for young children with autism spectrum disorder. AB - BACKGROUND: Recent estimates concerning the prevalence of autistic spectrum disorder are much higher than those reported 30 years ago, with at least 1 in 400 children affected. This group of children and families have important service needs. The involvement of parents in implementing intervention strategies designed to help their autistic children has long been accepted as helpful. The potential benefits are increased skills and reduced stress for parents as well as children. OBJECTIVES: The objective of this review was to determine the extent to which parent-mediated early intervention has been shown to be effective in the treatment of children aged 1 year to 6 years 11 months with autistic spectrum disorder. In particular, it aimed to assess the effectiveness of such interventions in terms of the benefits for both children and their parents. SEARCH STRATEGY: A range of psychological, educational and biomedical databases were searched. Bibliographies and reference lists of key articles were searched, field experts were contacted and key journals were hand searched. SELECTION CRITERIA: Only randomised or quasi-randomised studies were included. Study interventions had a significant focus on parent-implemented early intervention, compared to a group of children who received no treatment, a waiting list group or a different form of intervention. There was at least one objective, child related outcome measure. DATA COLLECTION AND ANALYSIS: Appraisal of the methodological quality of included studies was carried out independently by two reviewers. Differences between the included studies in terms of the type of intervention, the comparison groups used and the outcome measures were too great to allow for direct comparison. MAIN RESULTS: The results of this review are based on data from two studies. Two significant results were found to favour parent training in one study: child language and maternal knowledge of autism. In the other, intensive intervention (involving parents, but primarily delivered by professionals) was associated with better child outcomes on direct measurement than were found for parent-mediated early intervention, but no differences were found in relation to measures of parent and teacher perceptions of skills and behaviours. REVIEWER'S CONCLUSIONS: This review has little to offer in the way of implications for practice: there were only two studies, the numbers of participants included were small, and the two studies could not be compared directly to one another. In terms of research, randomised controlled trials involving large samples need to be carried out, involving both short and long term outcome information and full economic evaluations. Research in this area is hampered by barriers to randomisation, such as availability of equivalent services. PMID- 12535478 TI - Skin patch and vaginal ring versus combined oral contraceptives for contraception. AB - BACKGROUND: The delivery of combination contraceptive steroids from a skin patch or vaginal ring offers potential advantages over those administered via the traditional oral route. The skin patch and vaginal ring could possibly require a lower dose due to increased bioavailability and improved user compliance. OBJECTIVES: The review's objective is to compare the contraceptive efficacy, cycle control, compliance, and safety of the contraceptive skin patch versus combination oral contraceptives and the contraceptive vaginal ring versus combination oral contraceptives. SEARCH STRATEGY: We searched the computerized databases MEDLINE, Popline, Cochrane Controlled Trials Register, EMBASE, and LILACS for trials of the contraceptive patch or the contraceptive ring. We also searched the references of the publications identified for inclusion. We contacted the manufacturers of the skin patch and the contraceptive ring in an attempt to identify published or unpublished trials that we might have missed. SELECTION CRITERIA: All randomized controlled trials in any language comparing the combination contraceptive skin patch with a combination oral contraceptive or the combination contraceptive vaginal ring with a combination oral contraceptive. DATA COLLECTION AND ANALYSIS: Titles and abstracts identified from the literature searches were assessed for potential inclusion. Data were abstracted by both the primary and second reviewers to ensure accuracy and were entered into RevMan 4.1. Peto odds ratios with 95% confidence intervals were calculated for all outcomes. Six- and 13-cycle Kaplan-Meier pregnancy rates were also entered into "Additional Tables." No sensitivity analyses were conducted since the eligible trials used different comparison oral contraceptives. The trials were critically appraised by examining the following factors: the study design; blinding; randomization method; group allocation concealment; exclusions after randomization; loss-to follow-up; and early discontinuation. MAIN RESULTS: We found three trials of the skin patch and no eligible randomized controlled trials of the combination contraceptive vaginal ring. The six-cycle and 13-cycle Kaplan-Meier cumulative probabilities of pregnancy showed that the combination contraceptive skin patch was similar to the control combination oral contraceptive in contraceptive efficacy. One trial found that patch users discontinued early from the trial more often than oral contraceptive users (OR 1.6; 95% CI, 1.3-2.0) but a second trial found no differences in discontinuation between the groups (OR 2.6; 95% CI, 1.0 6.7). Patch users had more self-reported cycles of compliance than oral contraceptive users; the odds ratio of compliance was 2.1 (95% CI, 1.8-2.3). Patch users were more likely to report breast discomfort than oral contraceptive users with an odds ratio of 3.1 (95% CI, 2.3-4.2). The remaining commonly reported adverse events did not differ by group assignment. One trial found that women assigned to use the patch were more likely to discontinue due to adverse events than those assigned to use the oral contraceptive (OR 2.3; 95% CI, 1.6 3.3), but a second trial found no difference (OR 2.8; 95% CI, 0.7-11.3). Few serious adverse events occurred that were considered possibly or likely related to the use of the patch or oral contraceptive. REVIEWER'S CONCLUSIONS: The randomized controlled trials comparing a combination contraceptive skin patch to a combination oral contraceptive showed similar efficacy rates for the two methods. The patch group had better self-reported compliance than the oral contraceptive group. On the other hand, breast tenderness was more common among those randomized to the patch. The patch might lead to early discontinuation (overall and due to adverse events), but the trial results on this outcome were mixed. No conclusions can be drawn regarding the vaginal ring as no eligible trials were available. PMID- 12535479 TI - Interventions for treating collagenous colitis. AB - BACKGROUND: Collagenous colitis is a disorder that is recognized as a cause of chronic diarrhea. Treatment has been based mainly on anecdotal evidence. This review was performed to identify therapies for collagenous colitis that have been proven in randomized trials. OBJECTIVES: To determine effective treatments for patients with clinically active collagenous colitis. SEARCH STRATEGY: Relevant papers published between 1970 and October 2002 were identified via the MEDLINE, PUBMED, and EMBASE databases. Manual searches from the references of identified papers, as well as review papers on collagenous or microscopic colitis were performed to identify additional studies. Abstracts from major gastroenterological meetings were searched to identify research submitted in abstract form only. Finally, the Cochrane Controlled Trials Register and the Cochrane Inflammatory Bowel Disease Group Specialized Trials Register were searched for other studies. SELECTION CRITERIA: Four randomized trials were identified. One trial studied bismuth subsalicylate (published in abstract form only), and 3 trials (1 published in abstract form only) studied budesonide in the therapy of collagenous colitis. DATA COLLECTION AND ANALYSIS: Data were extracted independently by each author onto 2x2 tables (treatment versus placebo and response versus no response). For therapies assessed in one trial only, p-values were derived using the chi-square test. For therapies assessed in more than one trial, summary test statistics were derived using the Peto odds ratio and 95% confidence intervals. Data were combined for analysis only if the outcomes were sufficiently similar in definition. MAIN RESULTS: There were 9 patients with collagenous colitis in the trial studying bismuth subsalicylate (nine 262 mg tablets daily for 8 weeks). Those randomized to active drug were more likely to have clinical (p=0.003) and histological (p=0.003) improvement than those assigned to placebo. A total of 94 patients were enrolled in 3 trials studying budesonide (9 mg daily for 6 to 8 weeks). The pooled odds ratio for clinical response to treatment with budesonide was 12.32 (95% CI 5.53-27.46), with a number needed to treat of 2 patients. There was significant histological improvement with treatment in all 3 trials studying budesonide therapy. REVIEWER'S CONCLUSIONS: Budesonide is effective in the treatment of collagenous colitis. The evidence for bismuth subsalicylate is weaker, but still important. The roles of these and other therapies in inducing or maintaining remission (as opposed to clinical or histological improvement) of collagenous colitis are unknown. PMID- 12535480 TI - Elective delivery of women with a twin pregnancy from 37 weeks' gestation. AB - BACKGROUND: The optimal timing of birth for women with an otherwise uncomplicated twin pregnancy at term is uncertain, with clinical support for both elective delivery at 37 weeks, as well as expectant management (awaiting the spontaneous onset of labour). OBJECTIVES: To assess a policy of elective delivery from 37 weeks' gestation compared with an expectant approach for women with an otherwise uncomplicated twin pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register (searched 8 July 2002), Cochrane Controlled Trials Register (The Cochrane Library, Issue 2, 2002) and PubMed (January 1966 to 8 July 2002). SELECTION CRITERIA: Randomized controlled trials with reported data which compared outcomes in mothers and babies who underwent elective delivery from 37 weeks' gestation in a twin pregnancy with outcomes in controls who were managed expectantly. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. Outcomes listed were collected from the identified trial. MAIN RESULTS: A single randomised controlled trial comparing elective induction of labour at 37 weeks for women with a twin pregnancy with expectant management was identified. A total of 36 women were recruited to the trial with 17 women allocated to the induction of labour group and 19 women to the expectant management group. For primary outcomes, there were no statistically significant differences between elective induction of labour and expectant management with regards to all caesarean births (relative risk (RR) 0.56, 95% confidence interval (CI) 0.16 to 1.90), caesarean birth for fetal distress (RR 0.37, 95% CI 0.02 to 8.53), or perinatal death (RR not estimable). For secondary outcomes, there were no statistically significant differences between the two interventions with regards to haemorrhage requiring blood transfusion (RR 0.37, 95% CI 0.02 to 8.53), meconium stained liquor (RR 0.10, 95% CI 0.01 to 1.77), Apgar score of less than seven at five minutes (RR not estimable), and infant birth weight less than 2500 grams (RR 0.95, 95% CI 0.49 to 1.82). REVIEWER'S CONCLUSIONS: The small trial identified was underpowered to detect the outcome measures of interest. Consequently, there are insufficient data available to support a practice of elective delivery from 37 weeks gestation for women with an otherwise uncomplicated twin pregnancy at term. PMID- 12535481 TI - Interventions for replacing missing teeth: surgical techniques for placing dental implants. AB - BACKGROUND: Many variations in the surgical technique for the placement of dental implants have been developed since the introduction of implant surgery into clinical practice. These include variations in the timing of implant placement in relation to the tooth removal, and variations in the way the recipient bone site is prepared, amongst others. OBJECTIVES: To test the null hypothesis of no difference in the success, function, morbidity, patient satisfaction and cost effectiveness of different surgical techniques for placing dental implants, against the alternative hypothesis of a difference. SEARCH STRATEGY: The Cochrane Oral Health Group Trials Register, the Cochrane Central Register of Controlled Trials, MEDLINE and EMBASE were searched. In addition, the bibliographies of review articles were checked for studies outside the handsearched journals and personal references were searched. 55 implant companies were also contacted. SELECTION CRITERIA: Randomised controlled clinical trials (RCTs) of implant surgical techniques. DATA COLLECTION AND ANALYSIS: Authors were contacted for details of randomisation and data and quality assessment was carried out (ME, PC). Data were independently extracted, in duplicate, by two reviewers (HW, PC). The Cochrane Oral Health Group's statistical guidelines were followed. MAIN RESULTS: Four RCTs (six publications) were suitable for inclusion in this review of the nine RCTs (11 publications) identified. Two different aspects of implant surgical technique were reported in these RCTs. These were, two versus four implants to support a mandibular overdenture and crestal versus vestibular incision for implant placement. At the patient level there were no statistically significant differences for any of these alternative techniques with respect to implant failures, marginal bone levels, morbidity or patient satisfaction. REVIEWER'S CONCLUSIONS: This review included studies evaluating the surgical techniques of two versus four implants to support a mandibular overdenture and crestal versus vestibular incision for implant placement. Based on the available results of RCTs, there is no strong evidence supporting superior success with one or other of the alternative techniques for either of these two aspects of surgical technique. These conclusions are based on a few RCTs for each aspect of surgical technique and some with relatively short follow-up periods and few patients. PMID- 12535482 TI - Reduced dietary salt for prevention of cardiovascular disease. AB - BACKGROUND: Restricting sodium intake in elevated blood pressure over short periods of time reduces blood pressure. Long term effects (on mortality, morbidity or blood pressure) of advice to reduce salt in patients with elevated or normal blood pressure are unclear. OBJECTIVES: To assess in adults the long term effects (mortality, cardiovascular events, blood pressure, quality of life, weight, urinary sodium excretion, other nutrients and use of anti-hypertensive medications) of advice to restrict dietary sodium using all relevant randomised controlled trials. SEARCH STRATEGY: The Cochrane Library, MEDLINE, EMBASE, bibliographies of included studies and related systematic reviews were searched for unconfounded randomised trials in healthy adults aiming to reduce sodium intake over at least 6 months. Attempts were made to trace unpublished or missed studies and authors of all included trials were contacted. There were no language restrictions. SELECTION CRITERIA: Inclusion decisions were independently duplicated and based on the following criteria: 1) randomisation was adequate; 2) there was a usual or control diet group; 3) the intervention aimed to reduce sodium intake; 4) the intervention was not multifactorial; 5) the participants were not children, acutely ill, pregnant or institutionalised; 6) follow-up was at least 26 weeks; 7) data on any of the outcomes of interest were available. DATA COLLECTION AND ANALYSIS: Decisions on validity and data extraction were made independently by two reviewers, disagreements were resolved by discussion or if necessary by a third reviewer. Random effects meta-analysis, sub-grouping, sensitivity analysis and meta-regression were performed. MAIN RESULTS: Three trials in normotensives (n=2326), five in untreated hypertensives (n=387) and three in treated hypertensives (n=801) were included, with follow up from six months to seven years. The large, high quality (and therefore most informative) studies used intensive behavioural interventions. Deaths and cardiovascular events were inconsistently defined and reported; only 17 deaths equally distributed between intervention and control groups occurred. Systolic and diastolic blood pressures were reduced at 13 to 60 months in those given low sodium advice as compared with controls (systolic by 1.1 mm Hg, 95% CI 1.8 to 0.4, diastolic by 0.6 mm hg, 95% CI 1.5 to -0.3), as was urinary 24 hour sodium excretion (by 35.5 mmol/ 24 hours, 95% CI 47.2 to 23.9). Degree of reduction in sodium intake and change in blood pressure were not related. People on anti hypertensive medications were able to stop their medication more often on a reduced sodium diet as compared with controls, while maintaining similar blood pressure control. REVIEWER'S CONCLUSIONS: Intensive interventions, unsuited to primary care or population prevention programmes, provide only minimal reductions in blood pressure during long-term trials. Further evaluations to assess effects on morbidity and mortality outcomes are needed for populations as a whole and for patients with elevated blood pressure. Evidence from a large and small trial showed that a low sodium diet helps in maintenance of lower blood pressure following withdrawal of antihypertensives. If this is confirmed, with no increase in cardiovascular events, then targeting of comprehensive dietary and behavioural programmes in patients with elevated blood pressure requiring drug treatment would be justified. PMID- 12535483 TI - Group-based parent-training programmes for improving emotional and behavioural adjustment in 0-3 year old children. AB - BACKGROUND: Emotional and behavioural problems in children are common. Research suggests that parenting has an important role to play in helping children to become adjusted, and that the first few months and years of a child's life are especially important in establishing patterns of emotional, cognitive and social functioning which will in turn influence the child's future development and in particular, their mental health. Parenting programmes may therefore have a role to play in improving the emotional and behavioural adjustment of infants and toddlers. OBJECTIVES: The objectives of this review are as follows: a) To establish whether group-based parenting programmes are effective in improving the emotional and behavioural adjustment of children less than three years of age b) To assess the role of parenting programmes in the primary prevention of emotional and behavioural problems. SEARCH STRATEGY: A range of biomedical and social science databases were searched including MEDLINE, EMBASE, CINAHL, PsychLIT, Sociofile, Social Science Citation Index, ASSIA, the Cochrane Library including SPECTR, CENTRAL, National Research Register (NRR) and ERIC. SELECTION CRITERIA: Only randomised controlled trials of group based parenting programmes were included, and studies that had used at least one standardised instrument to measure emotional and behavioural adjustment. DATA COLLECTION AND ANALYSIS: The treatment effect for each outcome in each study was standardised by dividing the mean difference in post-intervention scores for the intervention and treatment group by the pooled standard deviation, to obtain an effect size. The results for each outcome in each study have been presented, with 95% confidence intervals. Where appropriate the results have been combined in a meta-analysis using a random effect model. MAIN RESULTS: Five studies were included in the review, and there was sufficient data from five studies to combine the results in a meta analysis. Meta-analyses were conducted for both parent-reports and independent assessments of children's behaviour. The result for parent reports shows a non significant result favouring the intervention group (ES -0.5, CI -1.06 to 0.08). The result for independent observations of children's behaviour shows a significant result favouring the intervention group (ES -0.54, CI -0.84 to 0.23). A meta-analysis of the limited follow-up data available shows a small non significant result favouring the intervention group (ES -0.24, CI -0.56 to 0.09). REVIEWER'S CONCLUSIONS: The findings of this review provide some support for the use of group-based parenting programmes to improve the emotional and behavioural adjustment of children under the age of 3 years. There is, however, insufficient evidence to reach any firm conclusions regarding the role that such programmes might play in the primary prevention of such problems. Furthermore, there is limited data available concerning the long-term effectiveness of these programmes, and the results from the two studies for which data was available produced borderline insignificant findings. Further research is required. PMID- 12535484 TI - Oral immunoglobulin for the treatment of rotavirus infection in low birth weight infants. AB - BACKGROUND: Rotavirus infection is the most common neonatal nosocomial viral infection. Epidemics with the newer P(6)G9 strains have been reported in neonatal units worldwide. These strains can cause severe symptoms in most infected infants. Infection control measures become necessary and the utilisation of hospital resources increase. Local mucosal immunity in the intestine to rotavirus is important in the resolution of infection and protection against subsequent infections. Boosting local immunity by oral administration of anti-rotaviral immunoglobulin preparations might be a useful strategy in treating rotaviral infections, especially in low birth weight babies. OBJECTIVES: To determine the effectiveness and safety of oral immunoglobulin preparations for the treatment of rotavirus infection in hospitalised low birthweight infants (birth weight less than 2500 gms) SEARCH STRATEGY: Electronic databases including The Cochrane Controlled Trials Register (The Cochrane Llibrary, Issue 2, 2002), MEDLINE (1966 June 2002), EMBASE (1980 - June 2002) and CINAHL (1982 - June 2002) were searched by the strategy outlined in the protocol. Science Citation Index search for all articles which referenced Barnes 1982 were searched. The proceedings of the Pediatric Academic Societies which were published in the journal, Pediatric Research, from 1991 were searched as well as abstracts of the doctoral dissertations and theses from 1960. All of the above search strategies were completed in June 2002. Authors prominent in the field were contacted for any unpublished articles and more information on published articles was sought. Reference lists of identified clinical trials and personal files were also reviewed. SELECTION CRITERIA: The criteria used to select studies for inclusion were: 1) DESIGN: randomised or quasi-randomised controlled trials 2) Hospitalised low birthweight infants with rotavirus infection 3) INTERVENTION: Oral immunoglobulin preparations compared to placebo OR no intervention 4) At least one of the following outcomes were reported: All cause mortality during hospital stay, mortality due to rotavirus infection during hospital stay, duration of diarrhoea, need for rehydration, duration of viral excretion, duration of infection control measures, length of hospital stay in days, recurrent diarrhoea or chronic diarrhoea DATA COLLECTION AND ANALYSIS: The two reviewers were to independently abstract data from eligible trials. No data analysis was possible at this point. MAIN RESULTS: No eligible randomised controlled trials were found. REVIEWER'S CONCLUSIONS: We found no randomised controlled trials which assessed the effectiveness or safety of oral immunoglobulin preparations for the treatment of rotavirus infections in hospitalised low birthweight infants. PMID- 12535485 TI - Interventions for the physical aspects of sexual dysfunction in women following pelvic radiotherapy. AB - BACKGROUND: Following pelvic radiotherapy (RT), a proportion of women experience problems related to sexual function, which are multifactorial in origin. The physical components relate to distortion of the perineum and vagina, which may occur as a result of surgery and/or radiotherapy and compromise sexual activity resulting in considerable distress. OBJECTIVES: The aim of this review was to evaluate the evidence for treatment options addressing the physical components of sexual dysfunction arising from pelvic radiotherapy as prevention or treatment of acute or late complications. SEARCH STRATEGY: The concepts used included synonyms for radiation therapy and brachytherapy and synonyms for the spectrum of physical aspects of sexual dysfunction in women. randomized. We searched the Cochrane Controlled Trials Register (CENTRAL), issue 1, 2002, MEDLINE 1966 to 2002, EMBASE 1980 to 2002, CANCERCD 1980 to 2002, Science Citation Index 1991 to 2002, CINAHL 1982 to 2002, as well as sources of grey literature. We also hand searched relevant textbooks and contacted experts in the field. SELECTION CRITERIA: Any study describing the therapeutic trial of a treatment to relieve the physical aspects of female sexual dysfunction which had developed following pelvic radiotherapy was considered. The quality of each study was then assessed by two reviewers independently to determine its suitability for inclusion in statistical analysis. DATA COLLECTION AND ANALYSIS: Thirty-two references met the inclusion criteria for the search but of these only four were suitable to be included for statistical analysis. MAIN RESULTS: The strongest evidence for benefit is the grade IC data in the topical oestrogens and benzydamine sections which describes the treatment of acute radiation vaginal changes. The use of vaginal dilators to prevent the development of vaginal stenosis is supported by grade IIC evidence. The value of hyperbaric oxygen therapy and surgical reconstruction is supported by the much weaker grade IIIC evidence in the form of case series. REVIEWER'S CONCLUSIONS: These findings reflect the quality of published data regarding interventions for this aspect of the management of radiation induced complications. Although there is grade IC evidence, these studies are not recent, the allocation concealment is unclear in the text, and overall there is a variable level of assessment of the response, emphasising the need for more studies to be conducted with improved designs to clarify the investigative process and support the final result. PMID- 12535486 TI - Closed reduction methods for treating distal radial fractures in adults. AB - BACKGROUND: Fracture of the distal radius is a common clinical problem, particularly in older white women with osteoporosis. Displaced fractures are usually reduced using closed reduction methods, which are non-surgical and generally comprise traction and manipulation, and the resulting position stabilised by external means, typically plaster cast immobilisation. OBJECTIVES: To examine the evidence for the relative effectiveness of different methods of closed reduction for displaced fractures of the distal radius in adults. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group specialised register (to July 2002), the Cochrane Controlled Trials Register (The Cochrane Library, Issue 3, 2002), MEDLINE (1966 to July week 4 2002), EMBASE (1988 to 2002 week 31), CINAHL (1982 to June week 4 2002), the UK National Research Register (Issue 2, 2002) and reference lists of articles. We also handsearched the British Volume of the Journal of Bone and Joint Surgery supplements (1996 onwards), and abstracts of the American Orthopaedic Trauma Association annual meetings. SELECTION CRITERIA: Randomised or quasi-randomised clinical trials evaluating different methods of closed reduction. We also included trials in which the use (or not) of anaesthesia could be classed as a co-intervention. DATA COLLECTION AND ANALYSIS: All trials judged as fitting the selection criteria by both reviewers were independently assessed by both reviewers for methodological quality. Data were extracted independently by one reviewer and checked by the other. Quantitative data are presented using relative risks or mean differences together with 95 per cent confidence limits. No pooling was possible. MAIN RESULTS: Three trials involving a total of 404, mainly female and older, patients with displaced fractures of the distal radius were included. These failed to assess functional outcome, and only one trial reported on complications. One trial found no statistically significant differences between mechanical reduction using finger trap traction and manual reduction in anatomical outcomes. One trial compared a novel method of manual reduction where the non-anaesthetised patient actively provided counter-traction versus traditional manual reduction under intravenous regional anaesthesia. While patients of the novel method group suffered more, yet not intolerable, pain during the reduction procedure, the latter was shorter in duration. No differences in anatomical outcome were detected. The third study compared mechanical reduction involving a special device without anaesthesia versus manual reduction under haematoma block (local anaesthesia). Less pain during the reduction procedure was recorded for the mechanical traction group. Both methods yielded similar radiological results. Fewer patients in the mechanical traction group had signs of neurological impairment, mainly finger numbness, at five weeks but this difference was not statistically significant by one year. REVIEWER'S CONCLUSIONS: There was insufficient evidence from comparisons tested within randomised trials to establish the relative effectiveness of different methods of closed reduction used in the treatment of displaced fractures of the distal radius in adults. Given the many unresolved questions over the management of these fractures, we suggest an integrated programme of research, which includes consideration of reduction methods, is the way forward. PMID- 12535487 TI - Homeopathy for dementia. AB - BACKGROUND: Dementia is a common illness in older people and has major implications for individuals with the disease, their carers and society. A meta analysis of population based studies in Europe found the prevalence of dementia in individuals over 65 to be 6.4%. Homeopathy (also spelt "homoeopathy") is a popular form of "complementary" or "alternative" treatment. Some studies have found evidence for efficacy of homeopathic treatment for some conditions, but any mechanism of action of the ultra molecular dilutions used in homeopathy is not comprehensible in terms of current scientific concepts. OBJECTIVES: To evaluate the effectiveness and safety profile of homeopathically prepared medications used in treating dementia, as established by randomized controlled trials. SEARCH STRATEGY: The trials were identified from a search of the Specialized Register of the Cochrane Dementia and Cognitive Improvement Group on 10 May 2002 using the terms alum*, homeop*, "nat sulph" and "natrum sulphate". This Register contains records from all major medical databases. In addition the reviewers searched CISCOM, AMED and Hom-Inform. Leading homeopathic researchers, practitioners and manufacturers were also contacted. SELECTION CRITERIA: All types of randomized controlled trials with a sample size of more than 20 were considered. DATA COLLECTION AND ANALYSIS: The database searches resulted in one trial being identified. After examining the abstract, a paper copy was obtained and independently assessed for inclusion by RM and JW. MAIN RESULTS: There were no studies that fulfilled the criteria for inclusion and no data to present. REVIEWER'S CONCLUSIONS: In view of the absence of evidence it is not possible to comment on the use of homeopathy in treating dementia. The extent of homeopathic prescribing for people with dementia is not clear and so it is difficult to comment on the importance of conducting trials in this area. PMID- 12535488 TI - Occlusal adjustment for treating and preventing temporomandibular joint disorders. AB - BACKGROUND: There has been a long history of using occlusal adjustment in the management of temporomandibular disorders (TMD). It is not clear if occlusal adjustment is effective in treating TMD. OBJECTIVES: To assess the effectiveness of occlusal adjustment for treating TMD in adults and preventing TMD. SEARCH STRATEGY: We searched the Cochrane Oral Health Group's Trials Register (April 2002); the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library Issue 2, 2002); MEDLINE (1966 to 8th April 2002); EMBASE (1980 to 8th April 2002) and handsearched journals of particular importance to this review. Additional reports were identified from the reference lists of retrieved reports and from review articles of treating TMD. There were no language restrictions. Unpublished reports or abstracts were considered from the SIGLE database. SELECTION CRITERIA: All randomised or quasi-randomised controlled trials (RCTs) comparing occlusal adjustment to placebo, reassurance or no treatment in adults with TMD. The outcomes were global measures of symptoms, pain, headache and limitation of movement. DATA COLLECTION AND ANALYSIS: Data were independently extracted, in duplicate, by two reviewers, Holy Koh (HK) and Peter G Robinson (PR). Authors were contacted for details of randomisation and withdrawals and a quality assessment was carried out. The Cochrane Oral Health Group's statistical guidelines were followed and relative risk values calculated using random effects models where significant heterogeneity was detected (P<0.1). MAIN RESULTS: Over 660 trials were identified by the initial search. Six of these trials, which reported results from a total of 392 patients, were suitable for inclusion in the review. From the data provided in the published reports, symptom-based outcomes were extracted from trials on treatment. Data on incidence of symptoms were extracted from trials on prevention. Neither showed any difference between occlusal adjustment and control group. REVIEWER'S CONCLUSIONS: There is an absence of evidence, from RCTs, that occlusal adjustment treats or prevents TMD. Occlusal adjustment cannot be recommended for the management or prevention of TMD. Future trials should use standardised diagnostic criteria and outcome measures when evaluating TMD. PMID- 12535489 TI - Prophylactic intravenous antifungal agents to prevent mortality and morbidity in very low birth weight infants. AB - BACKGROUND: Invasive fungal infection is an increasingly common cause of mortality and morbidity in very low birth weight infants. As the diagnosis is often difficult, and treatment is often delayed, there is a need to assess whether antifungal prophylaxis is beneficial. OBJECTIVES: To assess whether prophylactic intravenous antifungal therapy reduces mortality and adverse neurodevelopmental outcomes in very low birth weight infants. SEARCH STRATEGY: We used the standard search strategy of the Cochrane Neonatal Review Group. This included searches of the Cochrane Controlled Trials Register (The Cochrane Library, Issue 3, 2002), MEDLINE (1966 - September 2002), EMBASE (1980 - September 2002), conference proceedings, and previous reviews. SELECTION CRITERIA: Randomised controlled trials that compared the effect of prophylactic intravenous antifungal therapy versus placebo, or no drug, or another antifungal agent, in very low birth weight infants. DATA COLLECTION AND ANALYSIS: We extracted the data using the standard methods of the Cochrane Neonatal Review Group, with separate evaluation of trial quality and data extraction by each author, and synthesis of data using relative risk and risk difference. The pre specified outcomes were death prior to hospital discharge, longer term neurodevelopment, incidence of invasive fungal infection, emergence of antifungal resistance, and adverse drug reactions. MAIN RESULTS: We identified two eligible trials enrolling a total of 203 infants. Both trials compared the effect of prophylactic intravenous fluconazole versus placebo. 29 of the infants recruited to the two studies died. Meta-analysis revealed a statistically significantly reduced risk of death prior to hospital discharge for the infants who received fluconazole prophylaxis: typical relative risk: 0.44 (95% confidence interval 0.21, 0.91); typical risk difference: -0.11 (95% confidence interval -0.21, 0.02); number needed to treat: 9 (95% confidence interval 5, 50). There were not any data on longer term outcomes. REVIEWER'S CONCLUSIONS: We have found some evidence that prophylactic intravenous fluconazole reduces mortality prior to hospital discharge in very low birth weight infants. The meta-analysis suggests that there will be one fewer death in every nine infants treated with this intervention, but the 95% confidence interval around this estimate of effect is wide. The longer term neurodevelopmental consequences for infants exposed to this intervention remain to be determined. It will be important to identify any subgroups of very low birth weight infants that receive the most benefit from this intervention. There is also a need for further data on the effect of the intervention on the emergence of organisms with stable antifungal resistance. PMID- 12535490 TI - Single versus double intrauterine insemination (IUI) in stimulated cycles for subfertile couples. AB - BACKGROUND: Intra uterine insemination (IUI), with or without controlled ovarian hyperstimulation (COH), is one of the treatment modalities offered to couples who have tried to conceive for at least one year (subfertile couples). It has been suggested that increasing the number of inseminations from one per cycle to two might increase the probability of conception. OBJECTIVES: To determine if there is a difference in live birth or pregnancy rates for subfertile couples using single or double intrauterine insemination in stimulated cycles. SEARCH STRATEGY: We searched the Cochrane Menstrual Disorders and Subfertility group trials register (searched 8 July 2002), the Cochrane Controlled Trials Register (Cochrane Library Issue 1, 2002), MEDLINE (January 1966 to July 2002), EMBASE (January 1988 to July 2002), SCIENCE Direct Database (January 1966 to July 2002), Confsci (January 1973 to 15 July 2002), Pascal (January 1984 to July 2002) and reference lists of articles. We also contacted researchers in the field. SELECTION CRITERIA: Randomised controlled, parallel trials of single versus double intrauterine inseminations in simulated cycles in subfertile couples. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. Study authors were contacted for additional information. MAIN RESULTS: Three studies involving 386 women were included. There was no data for the main outcome measure of live birth per couple or ongoing pregnancy rates, and none presented comparative data for adverse events. The results of two studies that reported pregnancy rate per couple did not show a significant effect of using double insemination (Peto OR 1.45, 95% CI 0.78-2.70). REVIEWER'S CONCLUSIONS: Based on the results of pregnancy rate per couple of two trials, double intrauterine insemination showed no significant benefit over single intrauterine insemination in the treatment of subfertile couples with husband semen. There are no meaningful data to offer advice regarding clinical practice on the basis of this review. PMID- 12535491 TI - Interventions for replacing missing teeth: different times for loading dental implants. AB - BACKGROUND: To minimize the risk of implant failure, osseointegrated oral implants are conventionally kept load-free during the healing period. During healing removable prostheses are used, however many patients find these temporary prostheses rather uncomfortable and it would be beneficial if the healing period could be shortened without jeopardizing implant success. Nowadays immediate and early loaded implants are commonly used in mandibles of good bone quality. It would be useful to know whether there is a difference in success rates between immediately or early loaded implants compared with conventionally loaded implants. OBJECTIVES: To test the null hypothesis of no difference in the clinical performance between osseointegrated implants loaded at different times one year after loading. SEARCH STRATEGY: The Cochrane Oral Health Group Trials Register, The Cochrane Central Register of Controlled Trials, MEDLINE and EMBASE were searched. Handsearching included several dental journals. Authors of all identified trials and 55 oral implant manufacturers were contacted to find unpublished RCTs. The last electronic search was conducted on 8 May 2002. SELECTION CRITERIA: All RCTs of root-form osseointegrated oral implants having a follow-up of one year comparing the same osseointegrated root-form oral implants loaded at different times. DATA COLLECTION AND ANALYSIS: Data were independently extracted, in duplicate, by two reviewers. Authors were contacted for details of randomization and withdrawals and a quality assessment was carried out. The Cochrane Oral Health Group's statistical guidelines were followed. MAIN RESULTS: Only three RCTs were identified and two trials including 68 patients in total were suitable for inclusion in this review. Implants have been either immediately loaded after insertion (two to three days), early loaded (six weeks) or conventionally loaded (three to eight months) in edentulous mandibles of adequate bone quality and shape. On a patient, rather than per implant basis, there were no statistically significant differences for prosthesis failures, implant failures and marginal bone loss on intra-oral radiographs. REVIEWER'S CONCLUSIONS: While it is possible to successfully load oral implants immediately after their placement in mandibles of adequate bone density and height of carefully selected patients, it is yet unknown how predictable this approach is. More well designed RCTs are needed to understand how predictable immediate and early loading are. Such trials should be simply designed and should be reported according to the CONSORT guidelines (http://www.consort-statement.org/). It is suggested that priority should be given to trials assessing the effectiveness of immediately loaded implants rather than early loaded ones. PMID- 12535492 TI - Short versus long duration infusions of paclitaxel for any adenocarcinoma. AB - BACKGROUND: Paclitaxel has become a standard drug used in a number of common cancers. At first long infusions were used to reduce the rate of inflow of the drug and as a result reduce the occurrence of hypersensitivity types of allergic reactions. Trials with shorter durations of infusion, and using a cocktail of anti-allergic drugs to prevent hypersensitivity reactions, some randomised, were begun. These were interpreted as showing that effectiveness of treatment was not lessened by a short infusion time. These studies also appeared to show that some important toxicities were less common with short infusions and that they were more convenient for the patient and the hospital. OBJECTIVES: To assess the effect of varying the duration of infusion of paclitaxel on its anti-cancer effectiveness and side-effects. SEARCH STRATEGY: Electronic searches of the Cochrane Gynaecological Cancer CRG, the Cochrane Register of Controlled Trials, MEDLINE, EmBase, CANCERLIT, PDQ, Meta-register (mRCT) and the M.D. Anderson Cancer Centre, GOG were carried out. Information from the manufacturer and authors of reports of studies was also acquired. SELECTION CRITERIA: The review was restricted to randomised controlled trials of single agent paclitaxel or paclitaxel with other drugs, where the only variable was the duration of paclitaxel infusion. The review only included patients with advanced adenocarcinoma. DATA COLLECTION AND ANALYSIS: Data was extracted by two independent reviewers and where there was disagreement this was resolved by discussion. Where possible the data was synthesised in a meta-analysis. MAIN RESULTS: Three hour paclitaxel infusions appear to result in a smaller fall in white blood cell count, less fever, infection and sore mouth than 24 hour infusions. In contrast, 24 hour infusions cause less nerve toxicity. Other side effects are not dependent on the duration of infusion. Evidence from individual trials suggesting efficacy may be slightly greater with 24 hour infusions is inconclusive. Combination of data from trials of different cancer sites in a meta analysis must be considered speculative, but the combined data also suggest that 24 hour infusions of paclitaxel may be slightly more effective. REVIEWER'S CONCLUSIONS: This review confirms that, apart from neurological effects, three hour infusion of paclitaxel causes significantly less side effects than 24 hour infusion. Insufficient data exists to state whether varying the duration of infusion has a significant effect on its anti-cancer effectiveness. Further study would be required to establish whether there genuinely is a significant difference in efficacy according to the duration of infusion of paclitaxel. PMID- 12535493 TI - Continuous ambulatory peritoneal dialysis (CAPD) versus hospital or home haemodialysis for end-stage renal disease in adults. AB - BACKGROUND: Renal replacement therapy (RRT) with dialysis and transplantation is the only means of sustaining life for patients with end-stage renal disease (ESRD). Although transplantation is the treatment of choice, the number of donor kidneys are limited and transplants may fail. Hence many patients require long term or even life-long dialysis. Continuous ambulatory peritoneal dialysis (CAPD) is an alternative to hospital or home haemodialysis for patients with ESRD. OBJECTIVES: To assess the benefits and harms of CAPD versus hospital or home haemodialysis for adults with ESRD. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (Cochrane Library, Issue 2, 2002), the Cochrane Renal Group's Specialised Register (May 2002), MEDLINE (1966 - May 2002), EMBASE (1980 May 2002), BIOSIS, CINHAL, SIGLE and NRR without language restriction. Reference lists of retrieved articles and conference proceedings were searched and known investigators and biomedical companies were contacted. SELECTION CRITERIA: Randomised controlled trials (RCTs) or quasi-RCTs comparing CAPD to hospital or home haemodialysis for adults with ESRD were to be included. DATA COLLECTION AND ANALYSIS: Two reviewers were to independently assess the methodological quality of studies. Data was to be abstracted from included studies onto a standard form by one reviewer and checked by another. Statistical analyses were to be performed using the random effects model and the results to be expressed as relative risk (RR) with 95% confidence intervals (95% CI). MAIN RESULTS: Despite extensive searching, no RCTs or quasi-RCTs were identified. REVIEWER'S CONCLUSIONS: Data are not available to allow conclusions to be drawn about the relative effectiveness of CAPD compared with hospital or home haemodialysis for adults with ESRD. Efforts should be made to start and complete adequately powered RCTs, which compare the different dialysis modalities. PMID- 12535494 TI - Short versus standard duration oral antibiotic therapy for acute urinary tract infection in children. AB - BACKGROUND: The optimal duration of oral antibiotic therapy for urinary tract infection (UTI) in children has not been determined. A number of studies have compared single dose therapy to standard therapy for UTI, with mixed results. A course of antibiotics longer than a single dose but shorter than the usual 7-10 days might decrease the relapse rate and still provide some of the benefits of a shortened course of antibiotics. OBJECTIVES: The objective of this review was to assess the benefits and harms of short-course (2-4 days) compared to standard duration (7-14 days) oral antibiotic treatment for acute UTI in children. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (Cochrane Library Issue 3, 2002) MEDLINE (1966 - September 2002) and EMBASE (1988 -September 2002) without language restriction. SELECTION CRITERIA: Randomised and quasi-randomised controlled trials comparing short-term (2-4 days) with standard (7-14 days) oral antibiotic therapy were selected if they studied children aged three months to 18 years with culture proven UTI. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. Statistical analyses were performed using the random effects model and the results expressed as relative risk (RR) with 95% confidence intervals (95% CI). MAIN RESULTS: Ten trials were identified in which 652 children with lower tract UTI were evaluated. There was no significant difference in the frequency of positive urine cultures between the short (2-4 days) and standard duration oral antibiotic therapy (7-14 days) for UTI in children at 0-10 days after treatment (eight studies: RR 1.06; 95% CI 0.64 to 1.76) and at one to 15 months after treatment (10 studies: RR 0.95; 95% CI 0.70 to 1.29). There was no significant difference between short and standard duration therapy in the development of resistant organisms in UTI at the end of treatment (one study: RR 0.57, 95% CI 0.32 to 1.01) or in recurrent UTI (three studies: RR 0.39, 95% CI 0.12 to 1.29). REVIEWER'S CONCLUSIONS: A 2-4 day course of oral antibiotics appears to be as effective as 7-14 days in eradicating lower tract UTI in children. PMID- 12535495 TI - Haemoglobin and haematocrit targets for the anaemia of chronic renal disease. AB - BACKGROUND: Anaemia affects 60-80% of patients with renal impairment, reduces quality of life and is a risk factor for early death. Treatment options are blood transfusion, erythropoietin (EPO) alpha or beta and darbepoetin alfa. Recently higher haemoglobin (Hb) and haematocrit targets have been widely advocated because of positive data from observational studies. However, higher targets may lead to access thrombosis and hypertension and are costly. OBJECTIVES: This review assesses the benefits and harms of low (Hb 100 g/L or HCT > 30%) targets in pre- and post-dialysis patients receiving any treatment for anaemia. SEARCH STRATEGY: We searched The Cochrane Renal Group specialised register (September 2002), Cochrane Controlled Trials Register (The Cochrane Library, Issue 3, 2002) MEDLINE (1966 - September 2002), EMBASE (1988 - September 2002) and reference lists of retrieved articles. SELECTION CRITERIA: Randomised controlled trials (RCTs) and quasi-RCTs comparing low Hb/HCT targets (Hb 100 g/dL) in patients with anaemia of chronic renal disease. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. Statistical analyses were performed using the random effects model and the results expressed as relative risk (RR) for dichotomous outcomes and weighted mean difference (WMD) for continuous outcomes, with 95% confidence intervals (95% CI). MAIN RESULTS: Fifteen trials were identified in which 2096 patients were included. Twelve trials (673 patients) compared placebo with EPO and three trials (1423 patients) compared two doses of EPO. Hb values of 100 g/L (obtained with low EPO doses) were associated with lower mortality compared to Hb values of 140 g/L or more (obtained with high EPO doses) in the population with chronic renal disease and cardiovascular impairment (two trials, 1379 patients: RR 0.82; 95% CI 0.68 to 0.99). Lower targets obtained with a placebo resulted in an increased risk for seizures (four trials, 219 patients: RR 5.25; 95% CI 1.13 to 24.34) as compared to higher targets reached with EPO treatment. Finally, there was a reduced risk for hypertensive episodes with lower Hb targets reached with a placebo as compared to higher targets reached with EPO (six trials, 387 patients: RR 0.50; 95% CI 0.33 to 0.76). Quality of life was not adequately evaluated in the studies. REVIEWER'S CONCLUSIONS: Lower Hb targets of 100 g/L were associated with a lower risk of death in the population with cardiovascular impairment and chronic renal disease as compared to Hb 140 g/L. Lower Hb targets (Hb < 100 g/L) were also significantly associated with an increased risk for seizures and a reduced risk of hypertension compared to Hb > 100 g/L. There is a need of more adequately powered, well-designed and reported trials in this area. In particular, randomised controlled trials comparing the benefits and harms of low (Hb < 100 g/L) versus intermediate (Hb 130 g/L) and high (Hb 140 g/L) targets in the pre-dialysis population with chronic renal disease are necessary. In fact, there is a large deficiency of trials in the pre-dialysis population. The new trials should focus on hard outcomes and also look at outcomes which were previously not studied adequately, such as seizures and quality of life, which is to be assessed with validated measures. PMID- 12535496 TI - Psychological therapies for the management of chronic and recurrent pain in children and adolescents. AB - BACKGROUND: An increasing number of children suffer with pain that lasts for six months or longer. Traditional treatment for such pain has been pharmacological and/or physical. Increasingly, following developments in the field of adult chronic pain management, psychological therapies are being employed to treat children with chronic or recurrent pain. OBJECTIVES: To assess the effectiveness of psychological therapies in treating chronic or recurrent pain in children and adolescents, and to test the null hypothesis that psychological therapies are no more effective than placebo, waiting list control or standard medical care. SEARCH STRATEGY: Electronic searches of the Cochrane Register of Randomised Controlled Trials, MEDLINE (1966-1999), Social Sciences Citation Index (1981 1999) and PsycLit (1974-1999) were made. RCTs were also sought in references of all identified studies, meta-analyses and reviews, and first authors and experts within the field were contacted. Date of the most recent search: December 1999. SELECTION CRITERIA: RCTs with at least five participants in each study arm which compared psychological therapies with placebo, waiting list or standard medical care for children or adolescents with chronic or recurrent pain were eligible for inclusion. DATA COLLECTION AND ANALYSIS: Data were inspected for heterogeneity. For homogeneous dichotomous data the odds ratio with 95% confidence interval were calculated on an intention to treat basis. MAIN RESULTS: Thirty papers were recovered, representing 28 RCTs. Of these, 18 were analysable and included a total of 808 patients, 438 of whom entered treatment conditions. Fifteen were trials of chronic or recurrent headache; two for recurrent abdominal pain; and one for sickle cell pain. Only pain experience data from 13 trials were meta analysable. Two meta-analyses were conducted. The first analysis of single treatments versus controls gave a pooled odds ratio of 8.83 (95% CI 4.33 to 18.03; z=5.98, P < 0.00001, df = 12 ). The second analysis (combined treatment versus control) produced a similar estimate: pooled odds ratio = 8.64 ( 95% CI = 4.13 to 18.07; z-5.73, P < 0.00001, df = 9 ). Both analyses indicate that psychological treatment is effective when compared with a pooled group of control conditions. From the pooled data set the NNT was 2.32 (95%CI 1.96 to 2.88). REVIEWER'S CONCLUSIONS: There is very good evidence that psychological treatments, principally relaxation and cognitive behavioural therapy, are effective in reducing the severity and frequency of chronic headache in children and adolescents. There is at present no evidence for the effectiveness of psychological therapies in attenuating pain in conditions other than headache, and little evidence for the effectiveness of psychological therapies in improving non-pain outcomes. PMID- 12535497 TI - Human menopausal gonadotropin versus recombinant follicle stimulation hormone for ovarian stimulation in assisted reproductive cycles. AB - BACKGROUND: hMG and recombinant FSH, have both been used successfully for controlled ovarian hyperstimulation in in vitro fertilization and embryo transfer (IVF-ET). OBJECTIVES: To compare the effectiveness of hMG with rFSH in ovarian stimulation protocols in IVF or ICSI treatment cycles. SEARCH STRATEGY: We searched the Cochrane Menstrual Disorders and Subfertility Group trials register (searched 3rd Jan 2002), PubMed, MEDLINE, Web of Science (all searched 1985 to May 15 2002), and reference lists of articles. We also contacted manufacturers and researchers in the field. SELECTION CRITERIA: Randomised trials comparing hMG with rFSH for ovarian stimulation in IVF or ICSI treatment for treatment of infertility in normogonadotrophic women. DATA COLLECTION AND ANALYSIS: The main outcome measure was ongoing pregnancy/live birth per woman. Secondary outcomes included total gonadotrophin dose used, cancellation, number of oocytes retrieved, implantation, clinical pregnancy per woman, multiple pregnancy, spontaneous abortion and ovarian hyperstimulation syndrome. Peto odds ratios (OR) for hMG relative to rFSH were calculated after testing for homogeneity of treatment effect across all trials. Analyses were performed separately for the three different GnRHa protocols used: (1) without GnRHa down-regulation, (2) with GnRHa down-regulation using a short protocol and (3) with GnRHa down-regulation using a long protocol. MAIN RESULTS: Eight trials that met the inclusion criteria could be identified. One trial did not use down-regulation, one trial used a short protocol and six trials used a long down-regulation protocol. In the one trial with non-down-regulated women and in the one trial that used a short down regulation protocol there was no evidence of a difference between hMG and rFSH in any clinical outcome. Data of four truly randomised trials in women down regulated using a long protocol could be pooled. There was no evidence of a difference between hMG and rFSH in ongoing pregnancy/live birth per woman (OR 1.27; 95% CI 0.98 to 1.64). Furthermore there was no clear difference on any of the secondary outcomes, although the clinical pregnancy rate per woman was of borderline significance in favour of hMG (summary OR 1.28; 95% CI 1.00 to 1.64). The other secondary outcomes were comparable for both gonadotrophins. REVIEWER'S CONCLUSIONS: For all three GnRHa protocols analysed there is insufficient evidence of a difference between hMG and rFSH on ongoing pregnancy or live birth. More large randomised trials are needed to estimate the difference between hMG and rFSH more precisely. Such trials should preferably (1) use a consistent long GnRHa protocol and (2) use a fixed dose of gonadotrophin such to prevent potentially subjective decisions of the clinician in dosing and (3) take live birth as primary endpoint. At this moment in time however, in prescribing gonadotrophins for ovarian hyperstimulation in IVF one should use the least expensive medication. PMID- 12535498 TI - Placebo treatment versus no treatment. AB - BACKGROUND: Placebo interventions are often believed to improve patient reported and observer reported outcomes, but this belief is not based on evidence from randomised trials that compare placebo with no treatment. OBJECTIVES: To assess the effect of placebo interventions. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (The Cochrane Library, issue 3, 1998), MEDLINE (Jan 1966 to Dec 1998), EMBASE (Jan 1980 to Dec 1998), Biological Abstracts (Jan 1986 to Dec 1998), PsycLIT (Jan 1887 to Dec 1998). Experts on placebo research were contacted and references in the included trials were read. SELECTION CRITERIA: Randomised placebo trials with a no-treatment control group investigating any health problem were included. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. Study authors were contacted for additional information. MAIN RESULTS: Outcome data were available in 114 out of 130 included trials, investigating 40 clinical conditions. Outcomes were binary in 32 trials (3795 patients) and continuous in 82 (4730 patients). We found no statistically significant pooled effect of placebo in studies with binary outcomes, relative risk 0.95 (95 per cent confidence interval 0.88 to 1.02). The pooled relative risk for subjective (patient reported) outcomes was 0.95 (0.86 to 1.05) and for objective (observer reported) outcomes 0.91 (0.80 to 1.04). There was statistically significant heterogeneity (P < 0.03), but no evidence of sample size bias (P = 0.56). We found an overall positive effect of placebo treatments in trials with continuous outcomes, standardised mean difference -0.28 (95 per cent confidence interval -0.38 to -0.19). The standardised mean difference for subjective outcomes was -0.36 (-0.47 to -0.25), whereas no statistically significant effect was found for objective outcomes, standardised mean difference -0.12 (-0.27 to 0.03). There was statistically significant heterogeneity (P < 0.001), and evidence of sample size bias (P = 0.05). There was no statistically significant effect of placebo interventions in eight out of nine clinical conditions investigated in three trials or more (nausea, relapse in prevention of smoking and depression, overweight, asthma, hypertension, insomnia and anxiety), but confidence intervals were wide. There was a modest apparent analgesic effect of placebo interventions, standardised mean difference -0.27 (-0.40 to -0.15), but also a substantial risk of bias. REVIEWER'S CONCLUSIONS: There was no evidence that placebo interventions in general have clinically important effects. A possible moderate effect on subjective continuous outcomes, especially pain, could not be clearly distinguished from bias. PMID- 12535499 TI - Noradrenergic agonists for acute traumatic brain injury. AB - BACKGROUND: Although there have been considerable gains in understanding the cascade of events that lead to secondary injury after traumatic brain injury (TBI), efforts to translate this understanding into new therapeutic, so-called neuroprotective, approaches have so far proven disappointing. Animal models suggest an alternative strategy: agents enhancing monoaminergic transmission, particularly amphetamines, have been shown to promote motor recovery from focal brain injury and it has been suggested that this might represent a complementary means of therapeutic intervention in the later post-injury phase. OBJECTIVES: To evaluate the evidence that amphetamines improve final outcome after traumatic brain injury. SEARCH STRATEGY: We searched MEDLINE, EMBASE, Science Citation Index, Cochrane Controlled Trials Register and the Cochrane Injuries Group's Specialised Register of Controlled Trials. Researchers and authors of published trials were also contacted. SELECTION CRITERIA: Randomised controlled trials comparing the use of a noradrenergic agonist (together with conventional non pharmacological rehabilitative therapy) versus conventional non-pharmacological rehabilitative therapy alone. DATA COLLECTION AND ANALYSIS: Two reviewers independently screened records, extracted data and assessed trial quality. MAIN RESULTS: Although there is a limited clinical literature addressing this topic, none of the studies identified fully meets inclusion criteria for this review. REVIEWER'S CONCLUSIONS: At present there is insufficient evidence to support the routine use of methylphenidate or other amphetamines to promote recovery from TBI. PMID- 12535500 TI - Steroids for improving recovery following tonsillectomy in children. AB - BACKGROUND: Tonsillectomy continues to be one of the most common surgical procedures performed worldwide. Despite advances in anesthetic and surgical techniques, post-tonsillectomy morbidity remains a significant clinical problem. OBJECTIVES: To assess the clinical efficacy of a single intra-operative dose of dexamethasone in reducing post-tonsillectomy morbidity. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (Issue 1, 2002), MEDLINE (from 1966 - February 2002), EMBASE (from 1974 - February 2002) and reference lists of relevant articles. We contacted leading experts for information on any relevant unpublished data. SELECTION CRITERIA: Randomized, double-blind, placebo controlled trials of a single dose of intravenous, intra-operative corticosteroid for pediatric patients (age < 18 years) who underwent tonsillectomy or adenotonsillectomy were included. DATA COLLECTION AND ANALYSIS: Data regarding the primary outcome measures and measurement tools were extracted by the first author from the published studies. Data regarding study design, patient ages, procedures performed, dose of corticosteroid and method of delivery, as well as methodologic quality were also recorded by the first author. When data were missing from the original publications, the authors were contacted for more information. Data analysis was performed with a random effects model, using the RevMan 4.1 software developed by the Cochrane Collaboration. MAIN RESULTS: Children receiving a single intra-operative dose of dexamethasone (dose range = 0.15 to 1.0 mg/kg; maximum dose range = 8 to 25 mg) were two times less likely to vomit in the first 24 hours than children receiving placebo (RR = 0.54, CI95 = 0.42, 0.69; p < 0.00001). Routine use in four children would be expected to result in one less patient experiencing post-tonsillectomy emesis (RD = -0.25, CI95 = -0.37, -0.13; p = 0.00004). Additionally, children receiving dexamethasone were more likely to advance to a soft/solid diet on post-tonsillectomy day 1 (RR = 1.69, CI95 = 1.02, 2.79; p = 0.04) than those receiving placebo. Due to missing data and varied outcome measurement tools, pain could not be meaningfully analyzed as a distinct outcome measure. REVIEWER'S CONCLUSIONS: The evidence suggests that a single intravenous dose of dexamethasone is an effective, relatively safe and inexpensive treatment for reducing morbidity from pediatric tonsillectomy. No adverse events attributable to dexamethasone were reported in these trials. Additionally, in our 10-year experience of routine use of a single intravenous dose of dexamethasone during pediatric tonsillectomy, there have been no attributable, adverse events. Lastly, we found no reports in the literature of complications from use of a single intravenous dose of corticosteroid during pediatric tonsillectomy. PMID- 12535501 TI - Corticosteroid injections for shoulder pain. AB - BACKGROUND: While many treatments, including corticosteroid injections in and around the shoulder, are advocated to be of benefit for shoulder pain, few are of proven efficacy. This review of corticosteroid injections for shoulder pain is one in a series of reviews of varying interventions for shoulder disorders. OBJECTIVES: To determine the efficacy and safety of corticosteroid injections in the treatment of adults with shoulder pain. SEARCH STRATEGY: MEDLINE, EMBASE, CINAHL, Central and Science Citation Index were searched up to and including June 2002. SELECTION CRITERIA: Randomised and pseudo-randomised trials in all languages of corticosteroid injections compared to placebo or another intervention, or of varying types and dosages of steroid injection in adults with shoulder pain. Specific exclusions were duration of shoulder pain less than three weeks, rheumatoid arthritis, polymyalgia rheumatica and fracture. DATA COLLECTION AND ANALYSIS: Trial inclusion and methodological quality was assessed by two independent reviewers according to predetermined criteria. Results are presented separately for rotator cuff disease, adhesive capsulitis, full thickness rotator cuff tear and mixed diagnoses, and, where possible, combined in meta-analysis. MAIN RESULTS: Twenty-six trials met inclusion criteria. The number, site and dosage of injections varied widely between studies. The number of participants per trial ranged from 20 to 114 (median 52 participants). Methodological quality was variable. For rotator cuff disease, subacromial steroid injection was demonstrated to have a small benefit over placebo in some trials however no benefit of subacromial steroid injection over NSAID was demonstrated based upon the pooled results of three trials. For adhesive capsulitis, two trials suggested a possible early benefit of intra-articular steroid injection over placebo but there was insufficient data for pooling of any of the trials. One trial suggested short-term benefit of intra-articular corticosteroid injection over physiotherapy in the short-term (success at seven weeks RR=1.66 (1.21, 2.28). REVIEWER'S CONCLUSIONS: Despite many RCTs of corticosteroid injections for shoulder pain, their small sample sizes, variable methodological quality and heterogeneity means that there is little overall evidence to guide treatment. Subacromial corticosteroid injection for rotator cuff disease and intra-articular injection for adhesive capsulitis may be beneficial although their effect may be small and not well-maintained. There is a need for further trials investigating the efficacy of corticosteroid injections for shoulder pain. Other important issues that remain to be clarified include whether the accuracy of needle placement, anatomical site, frequency, dose and type of corticosteroid influences efficacy. PMID- 12535502 TI - Splints/orthoses in the treatment of rheumatoid arthritis. AB - BACKGROUND: Splints/orthoses are often recommended to patients with rheumatoid arthritis (RA) to decrease pain, decrease swelling and/or prevent deformity. These orthoses include resting hand splints, wrist supports, finger splints and special shoes and shoe inserts. OBJECTIVES: To assess the effectiveness of splints/orthoses in relieving pain, decreasing swelling and/or preventing deformity and determine the impact of splints/orthoses on strength, mobility and function in people with RA. SEARCH STRATEGY: We searched MEDLINE, EMBASE, the PEDro data base and Current Contents up to January 2002, and the Cochrane Controlled Trials Register to Issue 4, 2001 using the search strategy developed by the Cochrane Collaboration. Unpublished studies were sought by hand searching conference proceeding and contacting key experts. SELECTION CRITERIA: All randomized control trials (RCTs) and controlled clinical trials (CCTs), case control and cohort studies comparing the use of specific orthoses against placebo, another active intervention (including another type of orthoses) or regular treatment were selected, according to an a priori protocol. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected the studies and abstracted data. The methodological quality of the RCTs and CCTs was assessed using a validated scale. MAIN RESULTS: Twelve papers reporting on 10 studies met the inclusion criteria. These studies dealt with the following: working wrist splints (5), resting hand and wrist splints (2), special shoes and insoles (3). There is evidence that wearing working wrist splints statistically significantly decreases grip strength and does not affect pain, morning stiffness, pinch grip, quality of life after up to 6 months of regular wear. We found no evidence that resting wrist and hand splints change pain, grip strength, Ritchie Index or number of swollen joints. However, patients who wore these splints for 2 months reported that they preferred use to non-use and padded resting splints to unpadded ones. The one study of special shoes provided evidence of significant benefits of wearing extra-depth shoes for 2 months including less pain on walking and stair climbing and more minutes pain free walking time. Extra-depth shoes with semi-rigid insoles provided better pain relief than extra-depth shoes alone when worn over 12 weeks. Posted insoles prevented progression of hallux valgus angle but did not affect pain or function. REVIEWER'S CONCLUSIONS: There is insufficient evidence to make firm conclusions about the effectiveness of working wrist splints in decreasing pain or increasing function for people with RA. Potential adverse effects such as decreased range of motion do not seem to be an issue, although some of these splints decrease grip strength and dexterity. Similarly, preliminary evidence suggests that resting hand and wrist splints do not seem to affect range of motion or pain, although patients preferred wearing a resting splint to not wearing one. There is evidence that extra-depth shoes and molded insoles decreases pain on weight-bearing activities such as standing, walking and stair-climbing. Posted insoles may be effective in preventing progression of hallux abductus angle but do not appear to have an impact on pain. PMID- 12535503 TI - Effects of low sodium diet versus high sodium diet on blood pressure, renin, aldosterone, catecholamines, cholesterols, and triglyceride. AB - BACKGROUND: One of the controversies in preventive medicine is, whether a general reduction in sodium intake can decrease the blood pressure of a population and thereby reduce cardiovascular mortality and morbidity. In recent years the debate has been extended by studies indicating that reducing sodium intake has effects on the hormone and lipid profile. OBJECTIVES: To estimate the effects of low sodium versus high sodium intake on systolic and diastolic blood pressure (SBP and DBP), plasma or serum levels of renin, aldosterone, catecholamines, cholesterol and triglycerides. SEARCH STRATEGY: "MEDLINE" and reference lists of relevant articles were searched from 1966 through December 2001. SELECTION CRITERIA: Studies randomising persons to low sodium and high sodium diets were included if they evaluated at least one of the above outcome parameters. DATA COLLECTION AND ANALYSIS: Two authors independently extracted the data, which were analysed by means of Review Manager 4.1. MAIN RESULTS: In 57 trials of mainly Caucasians with normal blood pressure, low sodium intake reduced SBP by -1.27 mm Hg (CI: -1.76; -0.77)(p<0.0001) and DBP by -0.54 mm Hg (CI: -0.94; -0.14) (p = 0.009) as compared to high sodium intake. In 58 trials of mainly Caucasians with elevated blood pressure, low sodium intake reduced SBP by -4.18 mm Hg (CI: -5.08; - 3.27) (p < 0.0001) and DBP by -1.98 mm Hg (CI: -2.46; -1.32) (p < 0.0001) as compared to high sodium intake. The median duration of the intervention was 8 days in the normal blood pressure trials (range 4-1100) and 28 days in the elevated blood pressure trials (range 4-365). Multiple regression analyses showed no independent effect of duration on the effect size. In 8 trials of blacks with normal or elevated blood pressure, low sodium intake reduced SBP by -6.44 mm Hg (CI: -9.13; -3.74) (p < 0.0001) and DBP by -1.98 mm Hg (CI: -4.75; 0.78) (p = 0.16) as compared to high sodium intake. The magnitude of blood pressure reduction was also greater in a single trial in Japanese patients. There was also a significant increase in plasma or serum renin, 304% (p < 0.0001), aldosterone, 322%, (p < 0.0001), noradrenaline, 30% (p < 0.0001), cholesterol, 5.4% (p < 0.0001) and LDL cholesterol, 4.6% (p < 0.004), and a borderline increase in adrenaline, 12% (p = 0.04) and triglyceride, 5.9% (p = 0.03) with low sodium intake as compared with high sodium intake. REVIEWER'S CONCLUSIONS: The magnitude of the effect in Caucasians with normal blood pressure does not warrant a general recommendation to reduce sodium intake. Reduced sodium intake in Caucasians with elevated blood pressure has a useful effect to reduce blood pressure in the short term. The results suggest that the effect of low versus high sodium intake on blood pressure was greater in Black and Asian patients than in Caucasians. However, the number of studies in black (8) and Asian patients (1) was insufficient for different recommendations. Additional long-term trials of the effect of reduced dietary sodium intake on blood pressure, metabolic variables, morbidity and mortality are required to establish whether this is a useful prophylactic or treatment strategy. PMID- 12535504 TI - Early versus deferred treatment for early stage multiple myeloma. AB - BACKGROUND: Early stage multiple myeloma (MM) represents about 20% of MM. Most of the patients are asymptomatic. Thus, it is far less dramatic than advanced disease and may require different treatment strategies. For these patients, it is not clear whether it is better to start chemotherapy right after the diagnosis or to delay the treatment until symptoms become obvious as the disease progresses. OBJECTIVES: To identify and synthesize all available research evidence on whether early treatment intervention results in improved clinical outcomes when compared with observation alone. The main outcomes of interest that were examined included mortality, disease progression, response rate, and toxicity of early treatment. SEARCH STRATEGY: Searches of the following electronic databases were undertaken: MEDLINE, EMBASE, CANCERLIT, LILLIACS and Cochrane Database of RCTs. We have recently compiled a comprehensive database of RCTs in myeloma. This search was updated and supplemented by hand-search of abstracts from main society meetings such as the ASH (American Society of Hematology), ASCO (American Society of Clinical Oncology), and EHA (European Haematology Association). In addition, we compared our list with a list of RCTs maintained by the Oxford Clinical Trial Service Unit. SELECTION CRITERIA: Randomized controlled trials (RCT) with a parallel design that compared early versus deferred treatment of patients with early stage multiple myeloma based on Durie-Salmon (D-S) staging system. We also considered those trials that did not define early stage myeloma according to D-S staging system, but enrolled patients according to clinical uncertainty about the benefits of immediate intervention. DATA COLLECTION AND ANALYSIS: Data synthesis was performed for all studies and according to the defined quality criteria. The first reviewer and the contact reviewer of this proposal independently extracted data. Disagreement was resolved by consensus. Revman software (ver 4.1) was used to combine results from all studies and expressed as an overall odds ratio or Peto's Odds Ratio, with 95% confidence interval. MAIN RESULTS: Three trials were included with a total of 131 patients in each of the early treatment and deferred treatment groups. Early MM is asymptomatic stage I in these trials. All trials used standard Melphalan treatment but not stem cell transplantation. No statistically significant heterogeneity among the studies was detected. Beneficial effects of early treatment were seen in delay of myeloma progression (Peto's OR = 0.16, 95% CI: 0.09-0.29), and reduced vertebral compression (OR = 0.18, 95%CI: 0.02-1.59, NNT = 23, 95% CI: an NNT of 11, via infinity, to an NNH of 50). No significant effects on mortality and response rate were seen (Peto's OR = 1.11, 95% CI: 0.67-1.84, and OR = 0.63, 95% CI: 0.33-1.23, respectively). Early treatment may increase the risk of acute leukemia (Peto's OR = 3.20, 95% CI: 0.55-18.73, NNH = 44, 95% CI: an NNT of 63, via infinity, to an NNH of 15). REVIEWER'S CONCLUSIONS: Early treatment of early stage multiple myeloma inhibits disease progression, and may reduce vertebral compression. However, early treatment may increase the risk of acute leukemia. However, the data on vertebral compression and leukaemic transformation may not be interpretable due to very small numbers. Based on the current evidence, mortality and response rate are not significantly affected by introducing early treatment in the progression of myeloma. However, it is quite possible that the lack of beneficial effects of early intervention in myeloma is a false negative result due to the paucity of the existing evidence. In addition, data on quality of life and toxicity were sparsely reported adding to additional difficulties about management decisions in early stage myeloma. PMID- 12535505 TI - Day hospital versus admission for acute psychiatric disorders. AB - BACKGROUND: Inpatient treatment is an expensive way of caring for people with acute psychiatric disorders. It has been proposed that many of those currently treated as inpatients could be cared for in acute psychiatric day hospitals. OBJECTIVES: To assess the effects of day hospital versus inpatient care for people with acute psychiatric disorders. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (Cochrane Library, issue 4, 2000), MEDLINE (January 1966 to December 2000), EMBASE (1980 to December 2000), CINAHL (1982 to December 2000), PsycLIT (1966 to December 2000), and the reference lists of articles. We approached trialists to identify unpublished studies. SELECTION CRITERIA: Randomised controlled trials of day hospital versus inpatient care, for people with acute psychiatric disorders. Studies were ineligible if a majority of participants were under 18 or over 65, or had a primary diagnosis of substance abuse or organic brain disorder. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers and cross-checked. Relative risks and 95% confidence intervals (CI) were calculated for dichotomous data. Weighted or standardised means were calculated for continuous data. Day hospital trials tend to present similar outcomes in slightly different formats, making it difficult to synthesise data. Individual patient data were therefore sought so that outcomes could be reanalysed in a common format. MAIN RESULTS: Nine trials (involving 1568 people) met the inclusion criteria. Individual patient data were obtained for four trials (involving 594 people). Combined data suggested that, at the most pessimistic estimate, day hospital treatment was feasible for 23% (n=2268, CI 21 to 25) of those currently admitted to inpatient care. Individual patient data from three trials showed no difference in number of days in hospital between day hospital patients and controls (n=465, 3 RCTs, WMD -0.38 days/month CI -1.32 to 0.55). However, compared to controls, people randomised to day hospital care spent significantly more days in day hospital care (n=265, 3 RCTs, WMD 2.34 days/month CI 1.97 to 2.70) and significantly fewer days in inpatient care (n=265, 3 RCTs, WMD -2.75 days/month CI -3.63 to -1.87). There was no significant difference in readmission rates between day hospital patients and controls (n=667, 5 RCTs, RR 0.91 CI 0.72 to 1.15). For patients judged suitable for day hospital care, individual patient data from three trials showed a significant time-treatment interaction, indicating a more rapid improvement in mental state (n=407, Chi-squared 9.66, p=0.002), but not social functioning (n=295, Chi squared 0.006, p=0.941) amongst patients treated in the day hospital. Four of five trials found that day hospital care was cheaper than inpatient care (with cost reductions ranging from 20.9 to 36.9%). REVIEWER'S CONCLUSIONS: Caring for people in acute day hospitals can achieve substantial reductions in the numbers of people needing inpatient care, whilst improving patient outcome. PMID- 12535506 TI - Valproate for acute mood episodes in bipolar disorder. AB - BACKGROUND: Bipolar disorder is a common debilitating illness, characterised by acute affective episodes with full or partial inter-episode remission. Effective and acceptable treatment of acute episodes is required. Valproate has become a leading adjunctive and alternative mood stabilising treatment to lithium in bipolar disorder. OBJECTIVES: To determine the efficacy and acceptability of valproate in the treatment of acute episodes of bipolar disorder. SEARCH STRATEGY: The search included the Cochrane Collaboration Depression, Anxiety and Neurosis Controlled Trials Registrar (CCDANCTR), the Cochrane Controlled Clinical Trials Register (CCTR), reference lists of relevant papers and books, and contact with authors of trials, experts and pharmaceutical companies. SELECTION CRITERIA: Randomised controlled trials comparing valproate with placebo, other mood stabilisers and antipsychotic medication in the treatment of any bipolar affective episode. Participants were of both sexes, of all ages, with a diagnosis of bipolar affective disorder approximating to ICD 10 Code F31 and DSM IV 296. DATA COLLECTION AND ANALYSIS: Methodological quality was assessed independently by two reviewers blind to the authorship and source of papers. Ten randomised controlled trials were found comparing valproate with other interventions in mania. None was found examining its use in depression or mixed affective episodes. Data were extracted on the main outcome 'failure to respond by the end of the study' assessed by a less than 50% reduction in the Young Mania Rating Scale or the SADS-S mania scale. Three trials (316 participants) compared valproate with placebo. Three trials (158 participants) compared valproate with lithium. Two trials (363 participants) compared valproate with olanzapine. One trial (36 participants) compared valproate with haloperidol. Two trials (59 patients) compared valproate with carbamazepine. Acceptability of treatment was estimated using the outcome measure 'total number of subjects withdrawing from the study'. Three trials (321 patients) contributed to the comparison between valproate and placebo, two studies (144 patients) contributed to the comparison with lithium. One study (30 patients) provided data on this outcome in the comparison between valproate and carbamazepine. Pooled relative risks (with 95% confidence intervals) were calculated using fixed effect approaches. MAIN RESULTS: Valproate was more efficacious than placebo (RRR 38%; RR 0.62; 95% C.I. 0.51 to 0.77) in the treatment of mania. There was no significant difference between valproate and lithium (RRI 5%; RR 1.05; 95% C.I. 0.74-1.50) or between valproate and carbamazepine (RRR 34%; RR 0.66; 95% C.I. 0.38 to 1.16). Valproate was less effective than olanzapine (failure to achieve clinical response; RRI 25%; RR 1.25, 95% C.I. 1.01 to 1.54; average of 2.8 point less change on the Mania Rating Scale (95% CI 0.83 to 4.79). There were no significant differences in acceptability as measured by total number of subjects withdrawing from the study. There were significant differences in the side effect profiles of valproate and olanzapine, with more sedation and weight gain on olanzapine. REVIEWER'S CONCLUSIONS: There is consistent, if limited, evidence to suggest that valproate is an efficacious treatment for acute mania. Valproate may be less effective than olanzapine but may cause less sedation and weight gain. More well designed, randomised controlled trials investigating the relative efficacy and acceptability of valproate in the treatment of the full range of acute affective episodes occurring in bipolar disorder are required. PMID- 12535507 TI - Interventions for mucous membrane pemphigoid and epidermolysis bullosa acquisita. AB - BACKGROUND: Mucous membrane pemphigoid and epidermolysis bullosa acquisita are acquired autoimmune blistering diseases of the skin. Although they are rare, both can result in scarring of mucous membranes, which may lead to blindness and life threatening respiratory complications. OBJECTIVES: To assess the effects of treatments for mucous membrane pemphigoid and epidermolysis bullosa acquisita. SEARCH STRATEGY: Randomised Controlled Trials (RCTs) of patients with MMP or EBA were identified from MEDLINE and EMBASE from their inception to March 2000. The Cochrane Skin Group Specialised Register and the Cochrane Controlled Trials Register (CCTR) were last examined in February 2002. The bibliographies from identified studies were searched. The author who has conducted clinical trials in the field was contacted to identify unpublished trials. SELECTION CRITERIA: RCTs involving participants of any ages, and with a diagnosis confirmed by immunofluorescence. Where no RCTs were located, studies with other designs were considered. DATA COLLECTION AND ANALYSIS: Data were extracted from all included studies using a defined electronic data extraction protocol. Two reviewers evaluated the studies in terms of the inclusion criteria. The data from identified RCTs was extracted independently by three reviewers and subsequently checked for discrepancies. Any disagreements were resolved by discussion with each other and the fourth reviewer. Meta-analysis was not appropriate due to a lack of data. MAIN RESULTS: We found two small RCTs of MMP, both conducted in patients with severe eye involvement. The same author conducted both trials. In the first trial cyclophosphamide was superior to prednisone after six months of treatment; all 12 patients responded well to cyclophosphamide versus a good response in only five of 12 patients treated with prednisone (relative risk 2.40, 95% confidence interval 1.23 to 4.69). In the second trial all 20 patients treated with cyclophosphamide responded well to it after three months of treatment, but only 14 of 20 patients responded to the treatment with dapsone (relative risk 1.4, 95% confidence interval 1.07 to 1.90). We were not able to identify a RCT of therapeutic interventions in EBA. Thirty reports of uncontrolled studies of treatment for MMP involving five or more patients and 11 reports of treatment for EBA involving two or more patients were found, but were difficult to interpret. REVIEWER'S CONCLUSIONS: There is limited evidence (from two small trials) that severe ocular mucous membrane pemphigoid responds best to treatment with cyclophosphamide combined with corticosteroids, and that mild to moderate disease in most patients seems effectively suppressed by treatment with dapsone. It is difficult to make any treatment recommendations for EBA in the absence of reliable evidence sources. PMID- 12535508 TI - Radiofrequency denervation for neck and back pain. A systematic review of randomized controlled trials. AB - BACKGROUND: The diagnosis of cervical or lumbar zygapophyseal joint pain can only be made by using local anesthesia to block the nerves supplying the painful joint. There is a lack of effective treatment for chronic zygapophyseal joint pain or discogenic pain. Radiofrequency denervation appears to be an emerging technology, with substantial variation in its use between countries. OBJECTIVES: To assess the effectiveness of radiofrequency denervation for the treatment of musculoskeletal pain disorders. SEARCH STRATEGY: We searched MEDLINE, PsycLIT, and EMBASE from start to February 2002, plus the Cochrane Library 2002, Issue 2. The references of identified articles were checked and three experts in the field of radiofrequency treatment were consulted to identify studies we might have missed. SELECTION CRITERIA: Randomized controlled trials (RCTs) of radiofrequency denervation for musculoskeletal pain disorders, with no language or date restrictions. DATA COLLECTION AND ANALYSIS: Two reviewers selected RCTs that met predefined inclusion criteria, extracted the data, and assessed the main results and methodological quality of the selected trials, using standardized forms. Qualitative analysis was conducted to evaluate the level of scientific evidence. MAIN RESULTS: We found only nine articles, reporting on seven relevant RCTs. Six of the seven were considered to be high-quality. The selected trials included 275 randomized patients, 141 of whom received active treatment. One study examined cervical zygapophyseal joint pain, two cervicobrachial pain, three lumbar zygapophyseal joint pain, and one discogenic low-back pain. The study sample sizes were small, follow-up times short, and there were some deficiencies in patient selection, outcome assessments, and statistical analyses. The level of scientific evidence for the short-term effectiveness of radiofrequency denervation was limited for cervical zygapophyseal joint and cervicobrachial pain, and conflicting for lumbar zygapophyseal joint pain. There was limited evidence suggesting that intradiscal radiofrequency thermocoagulation was not effective for discogenic low-back pain. REVIEWER'S CONCLUSIONS: The selected trials provide limited evidence that radiofrequency denervation offers short-term relief for chronic neck pain of zygapophyseal joint origin and for chronic cervicobrachial pain; conflicting evidence on the short-term effect of radiofrequency lesioning on pain and disability in chronic low-back pain of zygapophyseal joint origin; and limited evidence that intradiscal radiofrequency thermocoagulation is not effective for chronic discogenic low-back pain. There is a need for further high-quality RCTs with larger patient samples and data on long term effects, for which current evidence is inconclusive. Furthermore, RCTs are needed in non-spinal indications where radiofrequency denervation is currently used without any scientific evidence. PMID- 12535509 TI - Non-invasive positive pressure ventilation for treatment of respiratory failure due to exacerbations of chronic obstructive pulmonary disease. AB - BACKGROUND: Non-invasive positive pressure ventilation (NPPV) is being used increasingly in the management of patients admitted to hospital with acute respiratory failure secondary to an exacerbation of chronic obstructive pulmonary disease (COPD). OBJECTIVES: To determine the effectiveness of NPPV in the management of patients with respiratory failure due to an acute exacerbation of COPD. SEARCH STRATEGY: An initial search was performed using the Cochrane Airways Group trials register and other relevant electronic databases. SELECTION CRITERIA: Randomised controlled trials comparing NPPV plus usual medical care versus usual medical care alone were selected. Trials needed to recruit adult patients admitted to hospital with respiratory failure due to an exacerbation of COPD and with PaCO2 > 6 kPa (45 mmHg). DATA COLLECTION AND ANALYSIS: Two reviewers independently selected articles for inclusion, evaluated methodological quality of the studies and abstracted the data. MAIN RESULTS: Eight studies were included in the review. NPPV resulted in decreased mortality (Relative Risk [RR] 0.41; 95% Confidence Intervals [CI] 0.26, 0.64), decreased need for intubation (RR 0.42; 95%CI 0.31, 0.59), reduction in treatment failure (RR 0.51; 95%CI 0.39, 0.67), rapid improvement within the first hour in pH (Weight Mean Difference [WMD] 0.03; 95%CI 0.02, 0.04), PaCO2 (WMD -0.40 kPa; 95%CI -0.78, -0.03) and respiratory rate (WMD -3.08 bpm; 95%CI -4.26, -1.89). In addition, complications associated with treatment (RR 0.32; 95%CI 0.18, 0.56) and length of hospital stay (WMD -3.24 days; 95%CI -4.42, -2.06) were also reduced in the NPPV group. REVIEWER'S CONCLUSIONS: Data from good quality randomised controlled trials permit NPPV to be recommended as the first line intervention, coupled with usual medical care, in all suitable patients with respiratory failure secondary to an acute exacerbation of COPD. A trial of NPPV should be considered early in the course of respiratory failure, and before severe acidosis ensures, as a means of avoiding endotracheal intubation, reducing mortality and treatment failure. PMID- 12535510 TI - Prophylactic antibiotic therapy for chronic bronchitis. AB - BACKGROUND: The use of prophylactic antibiotics to reduce the frequency and severity of acute exacerbations of chronic bronchitis is controversial. OBJECTIVES: To determine if prophylactic antibiotics reduce the frequency of exacerbations and/or days of disability in subjects with chronic bronchitis. SEARCH STRATEGY: We searched the Cochrane Airways Group Register of Clinical Trials and the bibliographies of relevant articles. SELECTION CRITERIA: Randomised controlled trials of prophylactic antibiotics in patients with chronic bronchitis and/or COPD were selected. DATA COLLECTION AND ANALYSIS: The eligibility of studies for inclusion was evaluated by three independent reviewers. MAIN RESULTS: Nine trials involving 1055 subjects were included in the analysis. All were performed before 1970. Concealment of allocation was assessed as clearly adequate in only 3 studies. The likelihood of having a exacerbation at any time during the course of the study was decreased with treatment (Relative Risk 0.91, 95% Confidence Intervals (CI) 0.84, 0.99). There was a small reduction in the number of exacerbations per patient per year with prophylactic antibiotics but this was not statistically significant ( Weighted Mean Difference (WMD) 0.15, 95%CI -0.34, 0.04 ). There was a modest but significant reduction of 22% in the number of days of disability per patient per month treated ( WMD -0.95, 95%CI -1.89 to - 0.01 ). A parallel reduction in the days of disability for each exacerbation (WMD -2.08, 95% CI -4.08 to -0.07) was seen. There was a small increase in adverse effects with antibiotics. REVIEWER'S CONCLUSIONS: Prophylactic antibiotics in chronic bronchitis / COPD have a small but statistically significant effect in reducing the days of illness due to exacerbations of chronic bronchitis. They do not have a place in routine treatment because of concerns about the development of antibiotic resistance and the possibility of adverse effects. The available data are over 30 years old, so the pattern of antibiotic sensitivity may have changed and there is a wider range of antibiotics in use. PMID- 12535511 TI - Options for self-management education for adults with asthma. AB - BACKGROUND: Asthma education and self-management are key recommendations of asthma management guidelines because they improve health outcomes. There are several different modalities for the delivery of asthma self-management education. OBJECTIVES: We evaluated programmes that: 1) Optimised asthma control through inhaled corticosteroid use by regular medical review or optimised asthma control by individualised written action plans 2) Used written self-management plans based on peak expiratory flow self-monitoring compared with symptom self monitoring 3) Compared different options for the delivery of optimal self management programmes. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and reference lists of articles. SELECTION CRITERIA: Randomised trials of asthma self-management education interventions in adults over 16 years of age with asthma. DATA COLLECTION AND ANALYSIS: Fifteen trials met the inclusion criteria. Trial quality was assessed and data were extracted independently by two reviewers. Study authors were contacted for confirmation. MAIN RESULTS: 1) Six studies compared optimal self-management allowing self adjustment of medications according to an individualised written action plan to adjustment of medications by a doctor. These two styles of asthma management gave equivalent effects for hospitalisation, ER visits, unscheduled doctor visits and nocturnal asthma. 2) Self-management using a written action plan based on PEF was found to be equivalent to self-management using a symptoms based written action plan in the six studies which compared these interventions. 3) Three studies compared self-management options. In one, that provided optimal therapy but tested the omission of regular review, the latter was associated with more health centre visits and sickness days. In another, comparing high and low intensity education, the latter was associated with more unscheduled doctor visits. In a third, no difference in health care utilisation or lung function was reported between verbal instruction and written action plans. REVIEWER'S CONCLUSIONS: Optimal self-management allowing for optimisation of asthma control by adjustment of medications may be conducted by either self-adjustment with the aid of a written action plan or by regular medical review. Individualised written action plans based on peak expiratory flow are equivalent to action plans based on symptoms. Reducing the intensity of self-management education or level of clinical review may reduce its effectiveness. PMID- 12535512 TI - A new connection: chaperones meet a mitochondrial receptor. AB - Cytosolic chaperones stabilize cellular proteins under stress conditions and protect nascent protein chains during normal growth. Recent data from Young et al. (2003) extend the function of chaperones by demonstrating that Hsp90 and Hsp70 specifically interact with the mitochondrial protein import receptor Tom70 at the outer membrane and are required for translocation of precursor proteins. PMID- 12535513 TI - Dissecting various ATP-dependent steps involved in proteasomal degradation. AB - Proteolysis by archaeal 20S proteasomes and the PAN (proteasome-activating nucleotidase) regulatory complex, a homolog of the eukaryotic 19S AAA ATPases, requires ATP hydrolysis through multiple steps. ATP hydrolysis, activated by binding of substrates to PAN, is utilized for substrate unfolding, gate opening of 20S proteasomes, and substrate translocation. PMID- 12535514 TI - Electrostatic fasteners hold the T cell receptor-CD3 complex together. AB - Assembly of the T cell receptor includes the formation of trimers stabilized by electrostatic interactions inside the membrane (Call et al., 2003). Such interactions can strongly stabilize subunit associations while permitting conformation changes during signaling. PMID- 12535515 TI - The forkhead transcription factor Foxo1: a possible link between obesity and insulin resistance. AB - The forkhead transcription factor Foxo1 has previously been shown to be a mediator of insulin action in liver and pancreas. New data from demonstrate that it also functions in adipose cells to couple insulin signaling to adipogenesis, which involves switching preadipocytes from proliferation to terminal differentiation. PMID- 12535516 TI - The unfolding story of polypeptide release factors. AB - Two recent cryo-EM reconstructions of the ribosome-bound release factor RF2 reveal an open, tri-lobed shape of RF2, in contrast to the comma-shaped molecule seen in the crystal structure. This indicates that RF2 undergoes a conformational change upon binding to the ribosome. Moreover, RF2 does not seem to be a molecular mimic of tRNA as is the case for elongation factor G. PMID- 12535517 TI - Akt phosphorylates the Yes-associated protein, YAP, to induce interaction with 14 3-3 and attenuation of p73-mediated apoptosis. AB - We have used an affinity purification method to identify substrates of protein kinase B/Akt. One protein that associates with 14-3-3 in an Akt-dependent manner is shown here to be the Yes-associated protein (YAP), which is phosphorylated by Akt at serine 127, leading to binding to 14-3-3. Akt promotes YAP localization to the cytoplasm, resulting in loss from the nucleus where it functions as a coactivator of transcription factors including p73. p73-mediated induction of Bax expression following DNA damage requires YAP function and is attenuated by Akt phosphorylation of YAP. YAP overexpression increases, while YAP depletion decreases, p73-mediated apoptosis following DNA damage, in an Akt inhibitable manner. Akt phosphorylation of YAP may thus suppress the induction of the proapoptotic gene expression response following cellular damage. PMID- 12535518 TI - Accelerated degradation of HMG CoA reductase mediated by binding of insig-1 to its sterol-sensing domain. AB - Sterols accelerate degradation of the ER enzyme 3-hydroxy-3-methylglutaryl CoA reductase (HMG CoA reductase), which catalyzes a rate-controlling step in cholesterol biosynthesis. This degradation contributes to feedback inhibition of synthesis of cholesterol and nonsterol isoprenoids. Here, we show that degradation of HMG CoA reductase is accelerated by the sterol-induced binding of its sterol-sensing domain to the ER protein insig-1. Accelerated degradation is inhibited by overexpression of the sterol-sensing domain of SREBP cleavage activating protein (SCAP), suggesting that both proteins bind to the same site on insig-1. Whereas insig-1 binding to SCAP leads to ER retention, insig-1 binding to HMG CoA reductase leads to accelerated degradation that is blocked by proteasome inhibitors. Insig-1 appears to play an essential role in the sterol mediated trafficking of two proteins with sterol-sensing domains, HMG CoA reductase and SCAP. PMID- 12535519 TI - A novel assay system implicates PtdIns(3,4)P(2), PtdIns(3)P, and PKC delta in intracellular production of reactive oxygen species by the NADPH oxidase. AB - Activated neutrophils assemble an NADPH oxidase enzyme complex to produce superoxide for microbial killing. Much of the initial oxidase assembly occurs on intracellular granules, followed by movement of the oxidase to phagolysosomes and the plasma membrane. We have developed a novel assay system using Streptolysin-O permeabilized neutrophils that recapitulates the initial intracellular activation process while maintaining the ultrastructural features of this granulocytic cell type. Using this system, we biochemically dissect molecular events and signaling pathways involved in NADPH oxidase assembly and demonstrate specific roles for PKC delta, PI(3,4)P(2)/PI(3,4,5)P(3), and PI(3)P in the PMA-dependent intracellular activation process. This system should be of great utility for the study of cell signaling events that regulate the intracellular production of reactive oxygen species by neutrophils. PMID- 12535520 TI - Structural determinants of integrin recognition by talin. AB - The binding of cytoplasmic proteins, such as talin, to the cytoplasmic domains of integrin adhesion receptors mediates bidirectional signal transduction. Here we report the crystal structure of the principal integrin binding and activating fragment of talin, alone and in complex with fragments of the beta 3 integrin tail. The FERM (four point one, ezrin, radixin, and moesin) domain of talin engages integrins via a novel variant of the canonical phosphotyrosine binding (PTB) domain-NPxY ligand interaction that may be a prototype for FERM domain recognition of transmembrane receptors. In combination with NMR and mutational analysis, our studies reveal the critical interacting elements of both talin and the integrin beta 3 tail, providing structural paradigms for integrin linkage to the cell interior. PMID- 12535521 TI - Identification of a new cryptochrome class. Structure, function, and evolution. AB - Cryptochrome flavoproteins, which share sequence homology with light-dependent DNA repair photolyases, function as photoreceptors in plants and circadian clock components in animals. Here, we coupled sequencing of an Arabidopsis cryptochrome gene with phylogenetic, structural, and functional analyses to identify a new cryptochrome class (cryptochrome DASH) in bacteria and plants, suggesting that cryptochromes evolved before the divergence of eukaryotes and prokaryotes. The cryptochrome crystallographic structure, reported here for Synechocystis cryptochrome DASH, reveals commonalities with photolyases in DNA binding and redox-dependent function, despite distinct active-site and interaction surface features. Whole genome transcriptional profiling together with experimental confirmation of DNA binding indicated that Synechocystis cryptochrome DASH functions as a transcriptional repressor. PMID- 12535522 TI - ATP hydrolysis by the proteasome regulatory complex PAN serves multiple functions in protein degradation. AB - To clarify the role of ATP in proteolysis, we studied archaeal 20S proteasomes and the PAN (proteasome-activating nucleotidase) regulatory complex, a homolog of the eukaryotic 19S ATPases. PAN's ATPase activity was stimulated similarly by globular (GFPssrA) and unfolded (casein) substrates, and by the ssrA recognition peptide. Denaturation of GFPssrA did not accelerate its degradation or eliminate the requirement for PAN and ATP. During degradation of one molecule of globular or unfolded substrates, 300-400 ATP molecules were hydrolyzed. An N-terminal deletion in the 20S alpha subunits caused opening of the substrate-entry channel and rapid degradation of unfolded proteins without PAN; however, degradation of globular GFPssrA still required PAN's ATPase activity, even after PAN-catalyzed unfolding. Thus, substrate binding activates ATP hydrolysis, which promotes three processes: substrate unfolding, gate opening in the 20S, and protein translocation. PMID- 12535523 TI - N-linked glycans direct the cotranslational folding pathway of influenza hemagglutinin. AB - For proteins that traverse the secretory pathway, folding commences cotranslationally upon translocation into the endoplasmic reticulum. In this study, we have comprehensively analyzed the earliest maturation steps of the model glycoprotein influenza hemagglutinin (HA). These steps include cleavage of the signal sequence, glycosylation, binding by the chaperones calnexin and calreticulin, and the oxidoreductase ERp57, and oxidation. Our results show that the molecular choreography of the nascent HA chain is largely directed by multiple glycans that are strategically placed to elicit the binding of lectin chaperones. These chaperones are recruited to specific nascent chain locations to regulate and facilitate glycoprotein folding, thereby suggesting that the positioning of N-linked glycans in critical regions has evolved to optimize the folding process in the cell. PMID- 12535524 TI - Structural basis of the ribosomal machinery for peptide bond formation, translocation, and nascent chain progression. AB - Crystal structures of tRNA mimics complexed with the large ribosomal subunit of Deinococcus radiodurans indicate that remote interactions determine the precise orientation of tRNA in the peptidyl-transferase center (PTC). The PTC tolerates various orientations of puromycin derivatives and its flexibility allows the conformational rearrangements required for peptide-bond formation. Sparsomycin binds to A2602 and alters the PTC conformation. H69, the intersubunit-bridge connecting the PTC and decoding site, may also participate in tRNA placement and translocation. A spiral rotation of the 3' end of the A-site tRNA around a 2-fold axis of symmetry identified within the PTC suggests a unified ribosomal machinery for peptide-bond formation, A-to-P-site translocation, and entrance of nascent proteins into the exit tunnel. Similar 2-fold related regions, detected in all known structures of large ribosomal subunits, indicate the universality of this mechanism. PMID- 12535525 TI - The critical role of the universally conserved A2602 of 23S ribosomal RNA in the release of the nascent peptide during translation termination. AB - The ribosomal peptidyl transferase center is responsible for two fundamental reactions, peptide bond formation and nascent peptide release, during the elongation and termination phases of protein synthesis, respectively. We used in vitro genetics to investigate the functional importance of conserved 23S rRNA nucleotides located in the peptidyl transferase active site for transpeptidation and peptidyl-tRNA hydrolysis. While mutations at A2451, U2585, and C2063 (E. coli numbering) did not significantly affect either of the reactions, substitution of A2602 with C or its deletion abolished the ribosome ability to promote peptide release but had little effect on transpeptidation. This indicates that the mechanism of peptide release is distinct from that of peptide bond formation, with A2602 playing a critical role in peptide release during translation termination. PMID- 12535526 TI - Coupled mRNA stabilization and translational silencing of cyclooxygenase-2 by a novel RNA binding protein, CUGBP2. AB - Cyclooxygenase-2 (COX-2) expression is translationally silenced in epithelial cells undergoing radiation-induced apoptosis. CUGBP2, a predominantly nuclear protein, is also rapidly induced in response to radiation and translocates to the cytoplasm. Antisense-mediated suppression of CUGBP2 renders radioprotection through a COX-2-dependent prostaglandin pathway, providing an in vivo demonstration of translation inhibition activity for CUGBP2. CUGBP2 binds to two sets of AU-rich sequences (AREs) located within the first sixty nucleotides of the COX-2 3' untranslated region (3'UTR). Upon binding, CUGBP2 stabilizes a chimeric luciferase-COX-2 3'UTR mRNA but inhibits its translation. These findings identify a novel paradigm for RNA binding proteins in facilitating opposing functions of mRNA stability and translation inhibition and reveal a mechanism for inhibiting COX-2 expression in cancer cells. PMID- 12535527 TI - The Q motif: a newly identified motif in DEAD box helicases may regulate ATP binding and hydrolysis. AB - SF1 and SF2 helicases have structurally conserved cores containing seven to eight distinctive motifs and variable amino- and carboxyl-terminal flanking sequences. We have discovered a motif upstream of motif I that is unique to and characteristic of the DEAD box family of RNA helicases. It consists of a 9 amino acid sequence containing an invariant glutamine. A conserved phenylalanine occurs 17 aa further upstream. Sequence alignments, site-specific mutagenesis, and ATPase assays show that this motif and the upstream phenylalanine are highly conserved, that they are essential for viability in the yeast Saccharomyces cerevisiae, and that they control ATP binding and hydrolysis in the yeast translation-initiation factor eIF4A. These results are consistent with computer studies of the solved crystal structures. PMID- 12535528 TI - Ligand-dependent nuclear receptor corepressor LCoR functions by histone deacetylase-dependent and -independent mechanisms. AB - LCoR (ligand-dependent corepressor) is a transcriptional corepressor widely expressed in fetal and adult tissues that is recruited to agonist-bound nuclear receptors through a single LXXLL motif. LCoR binding to estrogen receptor alpha depends in part on residues in the coactivator binding pocket distinct from those bound by TIF-2. Repression by LCoR is abolished by histone deacetylase inhibitor trichostatin A in a receptor-dependent fashion, indicating HDAC-dependent and independent modes of action. LCoR binds directly to specific HDACs in vitro and in vivo. Moreover, LCoR functions by recruiting C-terminal binding protein corepressors through two consensus binding motifs and colocalizes with CtBPs in the nucleus. LCoR represents a class of corepressor that attenuates agonist activated nuclear receptor signaling by multiple mechanisms. PMID- 12535529 TI - A shared surface of TBP directs RNA polymerase II and III transcription via association with different TFIIB family members. AB - The TATA box binding protein TBP is highly conserved and the only known basal factor that is involved in transcription by all three eukaryotic nuclear RNA polymerases from promoters with or without a TATA box. By mutagenesis and analysis on a selected set of four model pol II and pol III TATA box-containing and TATA-less promoters, we demonstrate that human TBP utilizes two modes to achieve its versatile functions. First, it uses a different set of surfaces on the conserved and structured TBP core domain to direct transcription from each of the four model promoters. Second, unlike yeast TBP, human TBP can use a shared surface to interact with two different TFIIB family members--TFIIB and Brf2--to initiate transcription by different RNA polymerases. PMID- 12535530 TI - The response regulator BvgA and RNA polymerase alpha subunit C-terminal domain bind simultaneously to different faces of the same segment of promoter DNA. AB - Examination of the binding of FeBABE-conjugated BvgA to the fha promoter of Bordetella pertussis has revealed that three dimers, formed by head-to-head association of monomers, bind one face of the DNA helix from the inverted-heptad primary binding site to the -35 region. The orientation of BvgA monomers within the dimers is the same as that recently demonstrated by X-ray crystallographic methods for a dimer of the C-terminal domain of NarL bound to DNA. Use of FeBABE conjugates of RNAP alpha subunit C-terminal domain showed that binding of this domain is linearly coincident with binding of the BvgA dimers, but to a different helical face. These results reveal a previously undescribed mode of interaction between RNAP alpha-CTD and a transcriptional activator. PMID- 12535531 TI - A cell cycle-regulated GATA factor promotes centromeric localization of CENP-A in fission yeast. AB - CENP-A, the centromere-specific histone H3 variant, plays a crucial role in organizing kinetochore chromatin for precise chromosome segregation. We have isolated Ams2, a Daxx-like motif-containing GATA factor, and histone H4, as multicopy suppressors of cnp1-1, an S. pombe CENP-A mutant. While depletion of Ams2 results in the reduction of CENP-A binding to the centromere and chromosome missegregation, increasing its dosage restores association of a CENP-A mutant protein with centromeres. Conversely, overexpression of CENP-A or histone H4 suppresses an ams2 disruptant. The intracellular amount of Ams2 thus affects centromeric nucleosomal constituents. Ams2 is abundant in S phase and associates with chromatin, including the central centromeres through binding to GATA-core sequences. Ams2 is thus a cell cycle-regulated GATA factor that is required for centromere function. PMID- 12535532 TI - Temporal regulation of topoisomerase IV activity in E. coli. AB - We isolated a mutant allele of dnaX, encoding the tau and gamma subunits of the DNA polymerase III holoenzyme, that causes extreme cell filamentation but does not affect either cell growth or DNA replication. This phenotype results from a defect in daughter chromosome decatenation during rapid growth. In these cells, ParC, one subunit of topoisomerase IV, no longer associated with the replication factory, as occurs in wild-type cells, and was instead distributed uniformly on the nucleoid; the distribution of ParE, the other subunit of topoisomerase IV, was unaffected. In addition, the majority of topoisomerase IV activity in synchronized cell populations was restricted to late in the cell cycle, when replication was essentially complete. These observations suggest that topoisomerase IV activity in vivo might be dependent on release of ParC from the replication factory. PMID- 12535533 TI - An ATR- and Cdc7-dependent DNA damage checkpoint that inhibits initiation of DNA replication. AB - We have analyzed how single-strand DNA gaps affect DNA replication in Xenopus egg extracts. DNA lesions generated by etoposide, a DNA topoisomerase II inhibitor, or by exonuclease treatment activate a DNA damage checkpoint that blocks initiation of plasmid and chromosomal DNA replication. The checkpoint is abrogated by caffeine and requires ATR, but not ATM, protein kinase. The block to DNA synthesis is due to inhibition of Cdc7/Dbf4 protein kinase activity and the subsequent failure of Cdc45 to bind to chromatin. The checkpoint does not require pre-RC assembly but requires loading of the single-strand binding protein, RPA, on chromatin. This is the biochemical demonstration of a DNA damage checkpoint that targets Cdc7/Dbf4 protein kinase. PMID- 12535534 TI - Progressive structural transitions within Mu transpositional complexes. AB - Assembly of the Mu transpososome is dependent on specific binding sites for the MuA transposase near the ends of the phage genome. MuA also contacts terminal nucleotides but only upon transpososome assembly, and base-specific recognition of the terminal nucleotides is critical for assembly. We show that Mu ends lacking the terminal 5 bp can form transpososomes, while longer DNA substrates with mutated terminal nucleotides cannot. The impact of the mutations can be suppressed by base mismatches near the end of Mu. Deletion of the flanking strands or mutation of the terminal nucleotides has differential effects on the cleavage and strand transfer reactions. These results show that the terminal nucleotides control the assembly and activation of transpososomes by influencing conformational changes around the active site. PMID- 12535535 TI - Excision of the Drosophila mariner transposon Mos1. Comparison with bacterial transposition and V(D)J recombination. AB - It has been proposed that the modern immune system has evolved from a transposon in an ancient vertebrate. While much is known about the mechanism by which bacterial transposable elements catalyze double-strand breaks at their ends, less is known about how eukaryotic transposable elements carry out these reactions. We have examined the mechanism by which mariner, a eukaryotic transposable element, performs DNA cleavage. We show that the nontransferred strand is cleaved initially, unlike prokaryotic transposons which cleave the transferred strand first. First strand cleavage is not tightly coupled to second strand cleavage and can occur independently of synapsis, as happens in V(D)J recombination but not in transposition of prokaryotic transposons. Unlike V(D)J recombination, however, second strand cleavage of mariner does not occur via a hairpin intermediate. PMID- 12535536 TI - Structural and dynamic functions establish chromatin domains. AB - Drosophila and mammalian proteins protect genes from heterochromatic repression in Saccharomyces cerevisiae by two different mechanisms. Factors termed genuine boundary activities (BAs) establish a structural, unidirectional bulwark against heterochromatin. In contrast, factors termed desilencing activities (DAs) act by the formation of a bidirectional, euchromatic island that blocks spreading of heterochromatin. The Drosophila boundary protein BEAF and, unexpectedly, the mammalian factor Sp1 exhibited a robust BA in yeast. In contrast, mammalian CTCF, Drosophila GAGA factor, yeast Gcn5p, and many mammalian transcription factors, although inactive as upregulators of nonsilenced genes, work as DAs. DAs but not BAs protect telomere-linked genes from silencing, presumably due to looping of telomeres and ensuing multidirectional silencing. The data demonstrate that "genetic autonomy" of chromatin domains is established by both passive and active mechanisms. PMID- 12535537 TI - Conformational flexibility underlies ubiquitin ligation mediated by the WWP1 HECT domain E3 ligase. AB - Ubiquitin ligases (E3) select proteins for ubiquitylation, a modification that directs altered subcellular trafficking and/or degradation of the target protein. HECT domain E3 ligases not only recognize, but also directly catalyze, ligation of ubiquitin to their protein substrates. The crystal structure of the HECT domain of the human ubiquitin ligase WWP1/AIP5 maintains a two-lobed structure like the HECT domain of the human ubiquitin ligase E6AP. While the individual N and C lobes of WWP1 possess very similar folds to those of E6AP, the organization of the two lobes relative to one another is different from E6AP due to a rotation about a polypeptide hinge linking the N and C lobes. Mutational analyses suggest that a range of conformations achieved by rotation about this hinge region is essential for catalytic activity. PMID- 12535538 TI - A conserved RING finger protein required for histone H2B monoubiquitination and cell size control. AB - Monoubiquitination of histone H2B is required for methylation of histone H3 on lysine 4 (K4), a modification associated with active chromatin. The identity of the cognate ubiquitin ligase is unknown. We identify Bre1 as an evolutionarily conserved RING finger protein required in vivo for both H2B ubiquitination and H3 K4 methylation. The RING domain of Bre1 is essential for both of these modifications as is Lge1 (Large 1), a protein required for cell size control that copurifies with Bre1. In cells lacking the euchromatin-associated histone variant H2A.Z, BRE1, RAD6, and LGE1 are each essential for cell viability, supporting redundant functions for H2B ubiquitination and H2A substitution in the formation of active chromatin. Notably, analysis of mutants demonstrates a function for Bre1/Lge1-dependent H2B monoubiquitination in the control of cell size. PMID- 12535539 TI - Bre1, an E3 ubiquitin ligase required for recruitment and substrate selection of Rad6 at a promoter. AB - Ubiquitination of histone H2B catalyzed by Rad6 is required for methylation of histone H3 by COMPASS. We identified Bre1 as the probable E3 for Rad6's role in transcription. Bre1 contains a C3HC4 (RING) finger and is present with Rad6 in a complex. The RING finger of Bre1 is required for ubiquitination of histone H2B, methylation of lysine 4 and 79 of H3 and for telomeric silencing. Chromatin immunoprecipitation experiments indicated that both Rad6 and Bre1 are recruited to a promoter. Bre1 is essential for this recruitment of Rad6 and is dedicated to the transcriptional pathway of Rad6. These results suggest that Bre1 is the likely E3 enzyme that directs Rad6 to modify chromatin and ultimately to affect gene expression. PMID- 12535540 TI - A heterotrimeric PCNA in the hyperthermophilic archaeon Sulfolobus solfataricus. AB - The sliding clamp, PCNA, of the archaeon Sulfolobus solfataricus P2 is a heterotrimer of three distinct subunits (PCNA1, 2, and 3) that assembles in a defined manner. The PCNA heterotrimer, but not individual subunits, stimulates the activities of the DNA polymerase, DNA ligase I, and the flap endonuclease (FEN1) of S. solfataricus. Distinct PCNA subunits contact DNA polymerase, DNA ligase, or FEN1, imposing a defined architecture at the lagging strand fork and suggesting the existence of a preformed scanning complex at the fork. This provides a mechanism to tightly couple DNA synthesis and Okazaki fragment maturation. Additionally, unique subunit-specific interactions between components of the clamp loader, RFC, suggest a model for clamp loading of PCNA. PMID- 12535541 TI - Field electron and ion emission from charged surfaces: a strategic historical review of theoretical concepts. AB - The field-electron (FE) and field-ion techniques directly observe and measure atomic-level surface processes that occur in very high electric fields. In theoretical terms, the high fields put large additional terms into Hamiltonians and free energies, and significantly modify many aspects of the surface physics and chemistry, as compared with the field-free situation. This paper presents a strategic review of the fundamental science of some of these high-field surface effects and processes, as developed in the context of the field electron and ion emission techniques. It outlines the main theoretical concepts developed, notes some twists of scientific history, and suggests useful contributions made to mainstream science. Topics covered are basic aspects of FE emission, surface field ionisation, localised field adsorption, charged surfaces theory, field-ion image contrast theory and associated imaging-gas kinetics, field evaporation, and aspects of the thermodynamics of charged surfaces. Despite many years of effort, important aspects of the theory remain incomplete. Some theoretical challenges are noted. PMID- 12535542 TI - Nanomaterials for field electron emission: preparation, characterization and application. AB - We review the current development in synthesis and deposition techniques of quasi one-dimensional nanomaterials and recent achievements in the study of the field electron emission properties of these materials. Major findings are given from our recent experimental study of the field electron emission characteristics of carbon nanotubes, copper sulfide and silicon carbide nanorods. The study reveals that the above nanomaterials are promising candidates as cold cathode electron emitters. Furthermore, we review the recent development of techniques for growing and depositing the above nanomaterials in device structures. Typical device structures and their performance are also shown. PMID- 12535543 TI - On the application of quantum transport theory to electron sources. AB - Electron sources (e.g., field emitter arrays, wide band-gap (WBG) semiconductor materials and coatings, carbon nanotubes, etc.) seek to exploit ballistic transport within the vacuum after emission from microfabricated structures. Regardless of kind, all sources strive to minimize the barrier to electron emission by engineering material properties (work function/electron affinity) or physical geometry (field enhancement) of the cathode. The unique capabilities of cold cathodes, such as instant ON/OFF performance, high brightness, high current density, large transconductance to capacitance ratio, cold emission, small size and/or low voltage operation characteristics, commend their use in several advanced devices when physical size, weight, power consumption, beam current, and pulse repletion frequency are important, e.g., RF power amplifier such as traveling wave tubes (TWTs) for radar and communications, electrodynamic tethers for satellite deboost/reboost, and electric propulsion systems such as Hall thrusters for small satellites. The theoretical program described herein is directed towards models to evaluate emission current from electron sources (in particular, emission from WBG and Spindt-type field emitter) in order to assess their utility, capabilities and performance characteristics. Modeling efforts particularly include: band bending, non-linear and resonant (Poole-Frenkel) potentials, the extension of one-dimensional theory to multi-dimensional structures, and emission site statistics due to variations in geometry and the presence of adsorbates. Two particular methodologies, namely, the modified Airy approach and metal-semiconductor statistical hyperbolic/ellipsoidal model, are described in detail in their present stage of development. PMID- 12535544 TI - Fowler-Nordheim theory for a spherical emitting surface. AB - In tests on a field emitter whose dimensions and work function were known, Fowler Nordheim (F-N) theory as usually stated for a planar emitter was found to give poor agreement with observations. The effect of curvature of the emitting surface has been modelled by including (a) non-linear variation of potential with distance from the surface; (b) the consequent changes in the exponent and pre exponential terms in the F-N expression for current; and (c) the variation of current density over the surface, modelled by an effective solid angle. Application of the resulting expression to the measured data gives estimates for apex radius which agree much more closely with the measured value than the value from planar theory does. PMID- 12535545 TI - Some comments on models for field enhancement. AB - To estimate the apex field-enhancement factor gamma(a)associated with a pointed protrusion on a flat planar surface, the simple physical models of a 'floating sphere at emitter-plane potential' and a 'hemisphere on a post' are often discussed. The corresponding mathematical expressions have the form: gamma(a)=m+h/rho, where rho is the sphere or hemisphere radius, h is its 'height above the emitter plane', and m is a constant variously taken as 0, 2 or 3. Recent numerical simulations for the 'hemisphere on a post' model, reported elsewhere by two of us (CJE and GV) and by Kokkaris, Modinos and Xanthakis, have shown that all of these simple formulae significantly overpredict gamma(a) if h/rho is large. This article first reexamines the basis of these simple formulae and confirms that they are less secure than is sometimes thought. The formulae reported elsewhere as fits to the numerical results are then quoted and compared with the simple formulae, and with the known exact analytical result for the 'hemi-ellipsoid on a plane' model. Discrepancies can be rationalised. Some general conclusions are drawn. PMID- 12535546 TI - Image potential between closely separated quantum-size film and a metal. AB - By theoretical analysis we have demonstrated that for degenerate films with a discrete electron energy spectrum (quantum-size (QS) films) the thickness dependences for the potential barrier height h(L) between a closely spaced QS film and a metal is quasi-oscillatory. This non-monotonic dependence of h(L) on the thickness d of the QS film, as well as the non-monotonic dependence of the main bulk characteristics of the QS film, can determine the dimensional dependence of the tunnel characteristics for spatially limited structures. PMID- 12535547 TI - Resonant field emission through amorphous diamond thin films (a model study). AB - A model for field emission through an amorphous diamond thin film with defects is constructed. Theoretical study shows that the emission is enhanced by attractive defects which would make the resonant emission observable for films with thickness of about 10nm. The emitted current density in typical parameters is calculated as functions of thickness, field strength and defect density. The energy distribution of emitted electrons is attained. PMID- 12535548 TI - A cold cathode lighting element prototype. AB - In this study, we present a prototype of lighting element in which a carbon containing field emission material is used as the cathode. The operating characteristics, i.e. current-voltage characteristics, current stability, luminance and lifetime of the lighting element are tested. By applying high voltage phosphor, a maximum brightness of 10000cd/m(2) has been recorded and a lighting element under lifetime test has been operating in DC driving mode for over 2500h without decay in emission current and brightness. These results imply that the prototype lighting element is suitable for commercial application. PMID- 12535549 TI - Field emission from individual multiwalled carbon nanotubes prepared in an electron microscope. AB - Individual multiwalled carbon nanotube field emitters were prepared in a scanning electron microscope. The angular current density, energy spectra, and the emission stability of the field-emitted electrons were measured. An estimate of the electron source brightness was extracted from the measurements. The results show that carbon nanotubes are promising candidates to replace existing sources in high-resolution electron beam instruments. PMID- 12535550 TI - Effect of carbon nanotube structural parameters on field emission properties. AB - Experimental studies were devoted to the effect of structural parameters, i.e., tube diameter and density, on the field electron emission characteristics of carbon nanotubes. Thermal chemical vapor deposition system was employed to synthesize carbon nanotubes. Nanotubes with different diameters and densities were obtained by adjusting the thickness of the iron (Fe) catalyst film. The morphologies of the Fe and carbon nanotube film were characterized by scanning electron microscopy respectively. Further field emission measurement confirmed that the tube diameter and density could significantly affect the electron emission properties of the carbon nanotube. Possible physical reasons for the effect are discussed. PMID- 12535551 TI - Field electron emission and nanostructural correlations for diamond and related materials. AB - Results are presented on a complex study of field electron emission (FEE) and structural correlations for nanocrystalline diamond and nitride films. It was found that all the samples studied showed similar dependences of the Fowler Nordheim work function and effective emitting area on the threshold emission field. Besides it was generally observed that FEE occurred at nanosized regions on the boundary of high and low conducting areas, and peaks of the emission intensity were associated with a lowered surface electron potential. Based on the experimental data, the following mechanism of low-FEE from the materials studied can be supposed. Electrons are transferred from the conducting channel into a vacuum through the low-dimensional region where the emission probability is high due to the quantum well effect. A physical model of the electron escape from a quantum well was analyzed. As follows from the estimations, the quantum size effect, being combined with a moderate field enhancement (the field enhancement factor beta=10-100), allows us to explain the observed variation in the energetic parameters for the samples studied. The function of the insulating grains is mainly to support the conducting channels in the sample body. Also, the grains can fulfill an additional function of the heat sink. PMID- 12535552 TI - Field emission from multiwall carbon nanotubes in controlled ambient gases, H2, CO, N2 and O2. AB - Adsorption and desorption on clean pentagons at a tip of multiwall carbon nanotube (MWNT) have been investigated by field emission microscopy (FEM) in an atmosphere of various gases, i.e., hydrogen, carbon monoxide, nitrogen and oxygen. A MWNT with clean surface which is obtained by heat treatment gives FEM patterns consisting of six bright pentagonal rings. Adsorbates are recognized as bright spots in the FEM pattern. They reside preferentially on the pentagonal sites where the strong electric field is concentrated, and bring about stepwise increase in the emission current. Heat treatment of the MWNT emitter at about 1300K allows adsorbates to desorb. After the desorption of hydrogen and nitrogen, the original clean surface with pentagons is recovered, while the tip structure is destroyed after the desorption of oxygen. PMID- 12535553 TI - Effects of hot electron emission on a low-conductivity tetracyanoethylene polymer layer including studies of the corrugation of the film surface. AB - The effect of strong field electron emission (FEE) on a tetracyanoethylene (TCNE) polymer layer was studied by Field Ion Microscopy (FIM) using TCNE and Ne as the imaging gases. The TCNE polymer was formed on each tungsten tip by radical polymerisation before FEE. The FIM images show field emission spots all over the surface of the tip. The FEM images show a random distribution of several field emission areas at the onset of FEE. After sometime at a current of about microA, there is a transition to higher currents at the same voltage, in which the electron emission pattern changes to have only one emitting area. After this transition, two different types of FIM images were observed, depending on the imaging gas that was used. Neon FIM images at low tip voltages show spots in the areas where the electron emission current was greatest, and at much higher voltages these images show emission from other areas with lower surface corrugation. However, the FIM images with TCNE as the imaging gas do not show any differences between the areas with and without electron emission. The FIM images remain as before FEE, which can be explained by the formation of a new polymer by the reaction of the surface layer with the imaging gas. It is assumed that chemically reactive fragments at the polymer/vacuum interface, which are needed for the polymerisation reaction, are formed by pyrolysis and sputtering processes during FEE. PMID- 12535554 TI - Measurements of the self-sustained enhancement of field emission by carbon fiber microemitters. AB - Two types of self-sustained enhancement in field emission by carbon fibers are described. In the first, the field is increased until the emission current switches from zero to between 1 and 10 microA. Next the field is reduced, but not so far that the current would drop. Then the current remains for several hours to several days, with transient increases from the 10 microA to between 14 and 22 microA. It is believed that the transients are caused by the activation of new microtips on the fiber surface. These effects were noted when the carbon fiber tip was mounted in a closed glass vacuum bulb pumped by barium getters, and also in a vacuum system using the combination of a molecular drag pump and ion pumps. The second type of enhancement occurs under ultrahigh vacuum conditions, during in situ thermal treatment of the carbon fiber tip while the emission current is about 2.5 microA. A specially built cathode assembly enables heating the tip to approximately 725 degrees C. After continuous heating at 570 degrees C for 20 to 35 h, the current suddenly increases to between 13 and 25 microA. This enhancement is reversible if the emitted current is kept at the newly increased value for at least 30 min. The current-voltage characteristics at several temperatures were recorded and analyzed. Similar field-forming phenomena were previously observed with Molybdenum and ZnO-W tips. PMID- 12535555 TI - Stabilization of carbon-fiber cold field-emission cathodes with a dielectric coating. AB - A comprehensive investigation has been carried out to determine the source of an inherent temporal instability in the spatial distribution and the electron emission current obtained from field-emitting carbon fiber tips. These instability effects were successfully overcome by coating the tip with a sub micron layer of dielectric epoxy resin coating. The influence of the coating thickness was studied and an optimum thickness of 0.2-0.3 microm that produced high emission stability was found. A large reduction in the intensity fluctuations of the emission image, at this coating thickness is demonstrated by using chart recorder traces in addition to slow scans of an optically monitored screen signal. The current-voltage (I-V) characteristics were obtained at a threshold field that is a few times lower than that of the uncoated tip. At low emission current levels linear F-N plots were obtained with a slope value lower than that of the uncoated emitter. The spatial distribution consisted of a very bright spot without any internal structure. The total energy distribution of the emitted electrons demonstrated a non-metallic behavior. The spectra obtained consisted of a single peak for low currents and a double peak for higher currents. The electron energy was measured relative to the Fermi level of tungsten and a spectral shift was shown to be a function of the current. Experiments have shown that the coated tips are not affected by the variations of pressure conditions down to 10(-6) mbar. These results suggest that a resin coated fiber tip offers superior performance to tungsten as a cold field emission electron source. Numerous improvements in the performance are underway. This includes a variety of polymeric coatings and more emissive carbon fibers. PMID- 12535556 TI - A high power density submicron electron beam source. AB - An electron-optical system (with a thermal field electron source-cathode) is introduced. The system is designed to form on the target surface submicron microprobes of a power density exceeding the ablation threshold ( approximately 10(8)W/cm(2)) of condensed media. The numerical modeling of electron paths carried out using a software package (TAU) developed by the authors has shown that the proposed electron-optical system is capable of forming microprobes of a power density comparable with that of modern high-power pulsed laser sources (up to approximately 10(11)W/cm(2)). PMID- 12535557 TI - Energy distributions of field emitted electrons from a multi-wall carbon nanotube. AB - Field emission energy distributions of electrons from one of the six pentagons located at the end of a multi-wall carbon nanotube have been measured by means of a high-resolution cylindrical energy analyzer. In a clean pentagon, the sub-peak was obtained at about 500 meV below the main peak, exhibiting a shift with increasing applied voltage. For electrons emitted from an adsorbate onto the pentagon, no fine structure was observed in the spectra. The broadening of the leading edge was also observed for both clean and adsorbed pentagon, indicating the field penetration into the nanotube due to its semimetallic nature. The full width at half-maximum was 280 meV at the applied voltage of 660 V and increased linearly with applied voltage. PMID- 12535558 TI - Field emission from diamond particles studied by scanning field emission microscopy. AB - Field emission properties from diamond particles (DPs) are studied. The DPs with thin chemically vapor deposited (CVD) diamond overcoat, dispersed onto metal substrate, essentially exhibit negative electron affinity (NEA). Field emission, approximately 1mA/cm(2) under a macroscopic electric field of 3.5kV/mm are observed. Microscopic electrical properties were studied by scanning tunneling microscopy/spectroscopy. Most parts of the DP surface exhibit narrow gap and p type characteristics. The localized regions, which have wide gap like bulk diamond properties, are randomly distributed near the top of DP. The field emission current distribution depicted by scanning field emission microscopy (SFEM) show that the electron emission is originating from a localized region on the selected DPs. We found, through SFEM measurement, some favorable field emission spots ("hot spots") where measured emission current is several orders higher than that of the other DPs ("normal spots"). Field emission spectroscopy (FES) results suggest that a poorly conducting layer is present along the electron path from the metal electrode to vacuum.We propose two models for field emission from "hot spots", which involve two main mechanisms. One is electron injection from the metal substrate to the DP, which is attributed to the electric field enhancement at intrinsic non-doped diamond (i-diamond) layer sandwiched between the metal substrate and the surface conductive layer (p-diamond) of the CVD diamond overcoat on the DP. The other is electron emission at the top site of NEA DP through the local i-diamond region or the depletion region of the p diamond, which is caused by the applied electric field. PMID- 12535559 TI - The application of carbon nanotubes in high-efficiency low power consumption field-emission luminescent tube. AB - In this report, details are given of our recent experimental study of using carbon nanotubes (CNTs) as cathode of the luminescent tubes. The CNT film is synthesized by thermal chemical vapor deposition. Two kinds of luminescent tubes that have different gaps have been fabricated. The luminescent tube with 0.1mm gap has a low threshold voltage of about 780V and high emission current of 300 microA when the gate voltage is 970V. The DC power consumption of the luminescent tube with a 0.1mm gap is approximately 1.12W. The results show that the CNTs are very good emission sources and suitable for application in the luminescent tube. PMID- 12535560 TI - Non-kinkwise field evaporation and kink relaxation on stepped W(112) surface. AB - Low-temperature field evaporation of the [111] steps on a W(211) surface was investigated by field ion microscopy (FIM). The atoms at the (112)[1;1;1] kink site are anomalously stable against field evaporation. This effect results in non kinkwise field evaporation near the kink of this type. The non-kinkwise field evaporating steps [211]<111> usually produce the reconstructed atomic chain with double-space arranged adatoms. The experimental results reveal atomic relaxation effects at steps and kinks. The normal to surface differential relaxation of the kink-site atoms was estimated by the geometrical method of indirect magnification and by simulation of the FIM image. PMID- 12535561 TI - STM observation of steps and terraces on tungsten (211) surface. AB - Thermally cleaned W(211) surface with 0.7 degrees miscut consists of (211) terraces separated by monoatomic steps. When the surface is exposed to oxygen and subsequently annealed at 1100-1900 K, the width of (211) terraces increases and multilayer steps are formed. Similar step bunching is observed during routine cleaning of the sample by annealing in oxygen and thermal flashing in ultra high vacuum. During such cleaning procedure islands of c(6 x 4) reconstruction are observed. PMID- 12535562 TI - On the temperature dependence of the electric characteristics and mass spectra of liquid metal alloy ion sources. AB - This paper reports on an extensive study of the characteristics of AuGeSi and CoNd liquid metal alloy ion sources (Focused Ion Beams From Liquid Metal Ion Sources, Research Studies Press, Taunton, Somerset, UK, 1991). Such characteristics include current-voltage curves and ion beam mass spectra. A careful investigation has been undertaken where the emitter temperature is the main variable experimental parameter. Theoretical models, or theoretical attempts in certain cases, tend to support the experimental results. The experimental results, in conjunction with existing theories, suggest that simply for reasons of self-consistency two mechanisms must be operative for the production of doubly charged ions from the AuGeSi source: direct field-evaporation and post ionisation. PMID- 12535563 TI - A new approach to gas field ion sources. AB - A new approach to gas field ion sources is described. It is based on a structure made by inserting a field emission tip inside a small diameter tube. The tube supplies gas to the tip from a high-pressure chamber into a high-vacuum chamber where ionization takes place. Comparison of projection electron and ion micrographs shows that ionization results from a field ionization process taking place at the very end of the tip. Emission currents in the 10nA range, for a few kV emission voltages, are obtained with various gases including neon, air and hydrogen. Lifetime experiments with H(2) show stable emission for days. PMID- 12535564 TI - Study on tip-substrate interactions by STM and APFIM. AB - Processes occurring at the interface of two materials coming in contact, separating or moving with respect to each other have been studied with the scanning tunnelling microscope (STM) and atom-probe (AP) field ion microscopy (APFIM). STM probe tips have been first characterised by field ion microscopy (FIM), brought into well-defined contact in the STM and afterwards inspected by time-of-flight AP. The results from mechanical contact and indentation experiments, showing material transfer and neck formation, are in reasonable good agreement with computer-based simulations on metal tip-surface interactions. PMID- 12535565 TI - Efficient sampling for three-dimensional atom probe microscopy data. AB - The best calculation of concentration profiles, isoconcentration surfaces or Gibbsian interfacial excesses from three-dimensional atom-probe microscopy data requires a compromise between spatial positioning error and statistical sampling error. For example, sampling from larger spatial regions decreases the statistical error, but increases the error in spatial positioning. Finding the appropriate balance for a particular calculation can be tricky, especially when the three-dimensional nature of the data presents an infinite number of degrees of freedom in defining surfaces, and when the statistical error is changing from one region of a sample to another due to differences in collection efficiency or atomic density. We present some strategies for approaching these problems, focusing on efficient algorithms for generating different spatial samplings. We present a unique double-splat algorithm, in which an initial, fine-grained sampling is taken to convert the data to a regular grid, followed by a second, variable width splat, to spread the effective sampling distance to any value desired. The first sampling is time consuming for a large dataset, but needs only be performed once. The second splat is done on a regular grid, so it is efficient, and can be repeated as many times as necessary to find the correct balance of statistical and positioning error. The net effect is equivalent to a Gaussian spreading of each data point, without the necessity of calculating Gaussian coefficients for millions of data points. We show examples of isoconcentration surfaces calculated under different circumstances from the same dataset. PMID- 12535566 TI - Time-of-flight atom probe measurements on Ni3Al and Co3W. AB - In this study, a VG FIM100 was taken into operation, consisting of a field-ion microscope (FIM), a time-of-flight atom probe (TOFAP) and an imaging atom probe. A tungsten specimen was used to calibrate the conversion of flight times to m/n values. The resulting relative mass resolution of the TOFAP was calculated to be m/Deltam approximately 500 FWHM. In time-of-flight measurements of homemade boron doped Ni-rich Ni(3)Al, a so-called ladder diagram was constructed from the evaporation data of a <001> pole. This ladder diagram revealed a very high degree of ordering in the alternating pure Ni and mixed Ni/Al planes: only 0.4% of the detected Al atoms were located on pure Ni planes. The number of null pulses to start a new plane was found to be much higher for Ni/Al planes (5 x 10(2)) than for Ni planes (1 x 10(2)). Moreover, the ladder diagram showed that boron was uniformly distributed through the matrix with nearly all boron found on pure Ni planes. The suggestion that boron preferentially settles on these planes would be supported by reports of a strong Ni-B bond, since on Ni planes, B atoms can be accommodated on octahedral interstitial sites surrounded by only Ni atoms. Finally, we performed time-of-flight measurements on Co(3)W. The ion species observed in these measurements included a wide variety of WN(n)(2+) ions, with 0 0.1), were greater for gilts fed the high energy, versus the low-energy, diet during gestation. Gilts that were fed the low energy diet during gestation consumed more feed during week 2 of lactation (P = 0.06). Our results suggest that altering the level of energy in the diets of gestating swine can influence circulating and milk concentrations of leptin, as well as feed consumption, during lactation. PMID- 12535587 TI - Effect of an asynchrony between ovulation and insemination on the results obtained after insemination with fresh or frozen sperm in rabbits. AB - The effects of the introduction of an 8-h asynchrony between ovulation and insemination on litter size components from rabbits were assessed. A total of 202 females belonging to a maternal line were used. Fresh and frozen sperm were used to perform the inseminations. Sperm was frozen with an extender composed of 1.75 M DMSO and 0.05 M sucrose. Four experimental groups were obtained depending on the type of sperm used (fresh or frozen) and on the moment that ovulation had been induced relative to the insemination (at the same time as insemination (t(0)) or 8 h before insemination (t(8))). Laparoscopy was performed on 12th day of pregnancy in pregnant females, and the ovulation rate, normal and total implanted embryos were noted. At kindling, total and live-born rabbits were noted. Results showed that better results were obtained after insemination with fresh semen than with frozen sperm (for females in the group t(0): 79% versus 61% fertility rate, 10.2 versus 6.4 normal implanted embryos and 8.1 versus 5.2 total number born, for fresh and frozen sperm, respectively). On the other hand, after the introduction of an 8-h asynchrony between ovulation and insemination, results were lower for both fresh (50% fertility rate, 7.5 normal implanted embryos and 5.7 total number born for the group of the asynchrony) and frozen sperm (31% fertility rate, 4.6 normal implanted embryos and 3.4 total number born for the group of the asynchrony). Although an approach between the moment of insemination and ovulation is justified when sperm survival could be compromised, results observed after the induction of an 8-h asynchrony were not those expected, perhaps due to the ageing of the oocytes before being fertilised, leading to both lack of fertilisation or early embryonic mortality. PMID- 12535588 TI - Fertility estimation: a review of past experience and future prospects. AB - Fertility has many components and stages which require that males and females be functionally capable of carrying out all critical stages if each generational reproductive cycle is to be completed. To accomplish this, the male must produce and ejaculate normal fertile sperm. The female must produce, store and ovulate normal fertilizable oocytes. Furthermore, the female must provide a reproductive system compatible with sperm transport, capacitation, and fertilization of the oocytes, embryo and fetal development, and finally birth of healthy young. Reproductive success or failure at several of these points can be estimated quantitatively on a population basis, and in a few situations on an individual basis. It is important that fertility estimates be determined accurately and with precision to be most useful to researchers and managers of animal enterprises. Many studies have underestimated the biological relationship of fertility to other traits because the estimates lacked precision. Many in vitro manipulations of sperm in artificial insemination, of gametes in various assisted reproductive technologies, and of embryos in embryo transfer are utilized in animal breeding programs. Accurate estimation of reproductive efficiency of these in vitro procedures also is important. Conditions surrounding different sets of fertility estimates almost certainly will be different. These conditions should be described as precisely as possible, and appropriate controls included in all experiments. When possible, experiments should be replicated over time and place to determine the repeatability of the various criteria used to estimate fertility and reproductive efficiency. Advances in genomic information and molecular biology should facilitate characterizing more fully inherent potential fertility of animals at birth. In vitro tests will improve, and automated techniques will facilitate making multiple determinations possible on a large scale. Reliability of fertility estimates will increase, with the potential for enhanced animal reproductive performance through more accurate selection, genetic engineering, and enlightened animal care. Simultaneously, it is important to recognize that prediction of future fertility is more hazardous than estimating fertility, as a completely new set of circumstances may occur which are not predictable. Because fertility estimation may be applied under a myriad of conditions, principles and factors affecting fertility will be emphasized in this review as being more useful than a compilation of numerical examples. PMID- 12535589 TI - Critical periods for foetal mortality in gilts identified by analysing the length distribution of mummified foetuses and frequency of non-fresh stillborn piglets. AB - The objective of this study was to investigate the timing of foetal mortality in gilts of a segregating F2 cross of Large White and Meishan pigs on the basis of the length distribution of mummified foetuses and the frequency of non-fresh stillborn piglets in order to establish whether critical periods for foetal mortality exist. All expelled conceptuses and placentae of 192 farrowing gilts with a normal health status were meticulously investigated to recover all mummified foetuses. The length of each mummified foetus was measured. The predicted number of foetuses present per gilt at the early foetal stage of gestation was calculated as the sum of numbers of mummified foetuses and non fresh stillborn, fresh stillborn and liveborn piglets. Foetal loss was calculated as the sum of mummified foetuses and non-fresh stillborn piglets. The average foetal mortality rate per gilt was 8.7%. In total 162 mummified foetuses were found (average 0.84 per litter), ranging in length from 0.4 to 33.0 cm. This indicates a range in foetal age at death of approximately 35-100 days. Although mummified foetuses of all lengths within the above mentioned range were found, relatively many had a length of less than 4 cm or of 10-21 cm. The total number of non-fresh stillborn piglets (i.e. late foetal deaths) was 58 (average 0.30 per litter). It can be concluded that foetal mortality occurred in these gilts throughout the period from day 35 to term, with relatively high incidences at the early foetal stage (days 35-40), shortly after mid-pregnancy (days 55-75) and after approximately day 100 of gestation. These three periods coincide with reported periods of change in porcine placental growth. PMID- 12535590 TI - Preliminary evidence that copper inhibits the degradation of DDT to DDE in pip and stonefruit orchard soils in the Auckland region, New Zealand. AB - Orchards (n=13) were sampled as part of a larger survey investigating agrichemical residues (pesticides and trace elements) in cropping soils in the Auckland region, New Zealand. SigmaDDT concentrations in orchard soils ranged from <0.03 to 24.41 mg kg(-1). DDT (o,p'- and p,p'-) comprised at least 40% of the SigmaDDT residues in 67% of orchards in which DDT residues were detected. There was a highly significant negative correlation (-0.924, P<0.001) between copper concentration (21-490 mg kg-1) and the ratio of DDE:DDT (0.4-5.2) in pip and stonefruit orchard soils. In further investigations involving five pip and stone fruit orchard sites and one grazing paddock it was found that soil respiration and the ratio of soil microbial carbon to soil carbon (%Cmic/Org-C) in orchard soils decreased with increasing copper concentration. These findings are consistent with the conclusion that elevated soil copper concentrations in pip and stone fruit orchard soils in the Auckland region may have reduced the ability of the indigenous soil microbial community to degrade DDT to DDE PMID- 12535591 TI - Temporal-spatial trends in heavy metal contents in sediment-derived soils along the Sea Scheldt river (Belgium). AB - The aim of this study was to survey the alluvial plains of the Sea Scheldt river in Belgium for the presence of old sediment-derived soils, and to appraise the heavy metal contamination at these sites. Historically, sediments of periodical dredging operations have been disposed in the alluvial plain without concern for the potential presence of contaminants. Up to 96% of the areas that were affected by sediment disposal (ca. 120 ha) was found to be polluted by at least one of the metals Cd, Cr, Zn or Pb. Concentrations of Cd, Cr and Zn were, in 10% of the cases, higher than 14, 1400 and 2200 mg/kg DM, respectively. Based on the Flemish decree on soil sanitation, Cu and Ni concentrations were of less environmental concern on any site. The pollution in the Sea Scheldt alluvial plain nevertheless is lower than for the Upper Scheldt alluvial plain. The sediment-derived soils in the most upstream part near Ghent were used for disposal of sediments from dredging operations elsewhere. Metal concentrations were explored and both spatial and temporal trend were analysed. The pollution levels encountered warrant caution as most of the soils affected by historical dredged sediment disposal are currently in use for pasture. PMID- 12535592 TI - Phosphate-induced metal immobilization in a contaminated site. AB - To assess the efficiency of P-induced metal immobilization in soils, a pilot scale field experiment was conducted at a metal contaminated site located in central Florida. Phosphate was applied at a P/Pb molar ratio of 4.0 with three treatments: 100% of P from H3PO4, 50% of P from H3PO4+ 50% of P from Ca(H2PO4)2, and 50% of P from H3PO4+5% phosphate rock in the soil. Approximately 1 year after P application, soil and plant samples were collected to determine mobility and bioavailability of selected metals (Pb, Zn, and Cu) using sequential extraction procedure and mineralogical characterization using X-ray diffraction (XRD) and scanning electron microscope-energy dispersive X-ray (SEM-EDX) analysis. Phosphorus distribution and soil pH effects were also evaluated. Phosphate was more effective in transforming soil Pb (to 53%) from the non-residual to the residual phase than soil Zn (to 15%) and soil Cu (to 13%). This was because Pb was immobilized by P via formation of an insoluble pyromorphite-like mineral in the surface and subsurface of the soil, whereas no phosphate mineral Zn or Cu was identified. While P amendment enhanced metal uptake in the roots of St. Augustine grass (Stenotaphrum secundatum), it significantly reduced metal translocation from root to shoot, especially Pb via formation of a pyromorphite-like mineral on the membrane surface of the root. A mixture of H3PO4 and phosphate rock was effective in metal immobilization, with less soil pH reduction and less soluble P. Although H3PO4 was effective in immobilizing Pb, its use should be limited to minimize soil pH reduction and potential eutrophication risk. PMID- 12535593 TI - Acid mine drainage arising from gold mining activity in Johannesburg, South Africa and environs. AB - The Witwatersrand region of South Africa is famous for its gold production and a major conurbation, centred on Johannesburg, has developed as a result of mining activity. A study was undertaken of surface and ground water in a drainage system in this area. Soils were also analysed from a site within the mining district. This study revealed that the ground water within the mining district is heavily contaminated and acidified as a result of oxidation of pyrite (FeS2) contained within mine tailings dumps, and has elevated concentrations of heavy metals. Where the water table is close to surface, the upper 20 cm of soil profiles are severely contaminated by heavy metals due to capillary rise and evaporation of the ground water. The polluted ground water is discharging into streams in the area and contributes up to 20% of stream discharge, causing a lowering of pH of the stream water. Much of the metal load is precipitated in the stream: Fe and Mn precipitate as a consequence of oxidation, while other heavy metals are being removed by co-precipitation. The oxidation of iron has created a redox buffer which controls the pH of the stream water. The rate of oxidation and of dilution is slow and the deleterious effect of the addition of contaminated water persists for more than 10 km beyond the source. PMID- 12535594 TI - The role of herbicides in the erosion of salt marshes in eastern England. AB - Laboratory studies and field trials were conducted to investigate the role of herbicides on saltmarsh vegetation, and their possible significance to saltmarsh erosion. Herbicide concentrations within the ranges present in the aquatic environment were found to reduce the photosynthetic efficiency and growth of both epipelic diatoms and higher saltmarsh plants in the laboratory and in situ. The addition of sublethal concentrations of herbicides resulted in decreased growth rates and photosynthetic efficiency of diatoms and photosynthetic efficiency of higher plants. Sediment stability also decreased due to a reduction in diatom EPS production. There was qualitative evidence that diatoms migrated deeper into the sediment when the surface was exposed to simazine, reducing surface sediment stability by the absence of a cohesive biofilm. Sediment loads on leaves severely reduced photosynthesis in Limonium vulgare. This, coupled with reduced carbon assimilation from the effects of herbicides, could have large negative consequences for plant productivity and over winter survival of saltmarsh plants. The data support the hypothesis that sublethal herbicide concentrations could be playing a role in the increased erosion of salt marshes that has occurred over the past 40 years. PMID- 12535595 TI - Accumulation of metals, trace elements and semi-volatile organic compounds on exterior window surfaces in Baltimore. AB - Organic films have been found to develop on window surfaces [Diamond et al., Environmental Science and Technology 34 (2000a) 2900]. The film contains organic compounds that are in dynamic equilibrium with the gas-phase in air, and organic and inorganic compounds and elements associated with deposited air particles. In this study, the exterior surfaces of windows were sampled in downtown and suburban Baltimore, Maryland. Higher concentrations of PCBs, PAH, metals and trace elements were found at downtown than a suburban site. PCBs in the films at downtown sites were dominated by penta and hexa homologue groups and PAH signatures resembled that of vehicle emissions. Twenty-six metals and trace elements were separated into two groups according to their enrichment factors (EF). Ag, Hg, Se, Sb and Zn had EF > 100, suggesting anthropogenic sources of these metals in the films; whereas Fe, Ca, Co, Cr and others had EF <10, suggesting a crustal origin. An unusual profile dominated by deca- and nona- PCBs and relatively higher concentrations of Ag, Hg and Zn may be attributable to emissions from a medical waste incinerator. Dry deposition fluxes of selected metals on windows were 1.4-94 times higher on windows with than without films, indicating that the film increases the dry deposition of particles and their associated chemicals. This implies that film development, which is associated with elevated VOC and SOC emissions to urban air, will increase film development that will, in turn, increase the accumulation of other atmospherically deposited constituents such as metals. PMID- 12535596 TI - Polybrominated diphenyl ethers in influents, suspended particulate matter, sediments, sewage treatment plant and effluents and biota from the Netherlands. AB - Polybrominated diphenyl ethers (PBDEs) have been determined in 133 samples of suspended particulate matter (SPM), sediments, sewage treatment plant (STP) influents and effluents, fish and mussels from various locations in The Netherlands, as a part of a large Dutch national study on estrogenic contaminants in the aquatic environment (LOES project). Some PBBs were also analysed but not found in any of the samples at detectable levels. PBDEs and PBBs were included in this study because indications of long term effects on the balance of endocrine systems were found in the literature. High concentrations of decaBDE (up to 4600 g/kg dry weight) were found in SPM from the Western Scheldt. These levels are possibly related to spillage during use of PBDEs in industries upstream the river Scheldt in Belgium. SPM was identified as an important carrier for higher brominated diphenyl ethers in the aquatic environment. DecaBDE was not found at detectable levels in flounder, bream and mussels. The bioaccumulation of decaBDE in these fish and shellfish samples is apparently limited. Lower brominated PBDE congeners (tetra/penta) were also found in the Western Scheldt as well as in the Rhine delta and the river Meuse, but in much lower concentrations than the decaBDE. In contrast with decaBDE, the tetra and pentaBDEs were found in biota. It was concluded that at least a small part of the PBDE can pass STPs. PMID- 12535597 TI - Polychlorinated naphthalenes in urban soils: analysis, concentrations, and relation to other persistent organic pollutants. AB - We determined the concentrations of 35 PCNs, 12 PCBs, and 20 PAHs in 49 urban topsoils under different land use (house garden, roadside grassland, alluvial grassland, park areas, industrial sites, agricultural sites) and in nine rural topsoils. The sums of concentrations of 35 PCNs (sigma35 PCNs) were <0.1-15.4 microg kg(-1) in urban soils and <0.1 to 0.82 microg kg(-1) in rural soils. The PCN, PCB, and PAH concentrations were highest at industrial sites and in house gardens. While rural soils receive PCNs, PCBs, and PAHs by common atmospheric deposition, there are site-specific sources of PCNs, PCBs, and PAHs for urban soils such as deposition of contaminated technogenic materials. The PCN, PCB, and PAH concentrations decreased from the central urban to the rural area. In the same order the contribution of lower chlorinated PCNs and PCBs increased because they are more volatile and subject to increased atmospheric transport. The PCNs 52+60, and 73 were more abundant in soil samples than in Halowax mixtures, indicating that combustion contributed to the PCN contamination of the soils. PMID- 12535598 TI - Trace element accumulation by moss and lichen exposed in bags in the city of Naples (Italy). AB - This paper presents the results of a bioaccumulation study of trace elements in the Naples urban area based on the use of the moss Sphagnum capillifolium (Ehrh.) Hedw. and the lichen Pseudevernia furfuracea (L.) Zopf exposed in bags in 23 sites. Moss and lichen bags were exposed for 4 months starting from the beginning of July 1999. Bags gathering was carried out after 10 weeks of exposure, at the end of the dry season, and after 17 weeks, during the wet season. The elements Al, As, Ca, Cd, Cr, Co, Cu, Fe, K, Mg, Mn, Mo, Ni, Pb, Ti, V and Zn were analysed by inductively coupled plasma-mass spectrometry ICP-MS in both biomonitors. For the majority of the elements the total amounts found in S. capillifolium were higher than in P. furfuracea whether considering the whole period of exposure or the weekly uptake. It was observed that there was a much greater difference in metal accumulation by P. furfuracea between the dry and wet seasons compared with S. capillifolium. In the wet period, the lichen seems to accumulate a larger quantity of metals. With the exception of Mn, trace element concentrations did not appear to be significantly affected by the washing away of rainfall. K loss during exposure suggested cell membrane damage in both organisms. For P. furfuracea the K leakage was limited to the dry period of exposure. A clear distinction between "lithophilic" and "anthropogenic" elements was achieved by cluster analysis. Significant correlations were found among Fe-Cu-Cr-Ni, Pb-Cd Co, V-Cr-Ni, Zn-Ni-Pb, suggesting a common source for each group of elements. PMID- 12535599 TI - Characterization of urban air quality using GIS as a management system. AB - Keeping the air quality acceptable has become an important task for decision makers as well as for non-governmental organizations. Particulate and gaseous emissions of pollutant from industries and auto-exhausts are responsible for rising discomfort, increasing airway diseases, decreasing productivity and the deterioration of artistic and cultural patrimony in urban centers. A model to determine the air quality in urban areas using a geographical information system will be presented here. This system permits the integration, handling, analysis and simulation of spatial and temporal data of the ambient concentration of the main pollutant. It allows the users to characterize and recognize areas with a potential increase or improvement in its air pollution situation. It is also possible to compute past or present conditions by changing basic input information as traffic flow, or stack emission rates. Additionally the model may be used to test the compliance of local standard air quality, to study the environmental impact of new industries or to determine the changes in the conditions when the vehicle circulation is increased. PMID- 12535600 TI - Relations between Scots pine needle element concentrations and decreased needle longevity along pollution gradients. AB - Scots pine (Pinus sylvestris L.) shoots were sampled along transects near one urban pollution source and two smelters. Needle Mg, P and K concentrations decreased from the second to the fourth age class linearly with needle survival along the urban pollution gradient. Still, over 80% of the average concentration of these nutrients remained in the fourth needle age class. Decreased needle longevity was closely related to the increased heavy metal concentrations near the smelters. Near the urban pollution source, it was related to the increased annual needle mass and the increased needle nutrient concentrations. Decreased Mn accumulation along with needle age was detected near all pollution sources. Leaching of Mn from needles and especially from soil as a cause of decreased needle concentrations is discussed. PMID- 12535601 TI - Ecotoxicological suitability of floodplain habitats in The Netherlands for the little owl (Athene noctua vidalli). AB - This study describes the actual risks of exposure to contaminants, which little owls (Athene noctua vidalli) face in Dutch river floodplains. The results indicate that PCBs pose a risk: not only are levels in little owls from floodplains higher than levels found in little owls from a reference site but the PCB patterns in owls from the floodplains also indicate induction of hepatic cytochrome P450 enzymes by dioxin like compounds, possibly PCBs. Of the heavy metals, only cadmium is thought to pose a risk in certain conditions, for example, when little owls are feeding only on earthworms over a prolonged period of time. The results do not indicate any effects on the occurrence of prey items of the little owl like for instance earthworm, beetles and shrews. Hence, it is not expected that little owls will be affected by diminishing prey availability due to contamination. PMID- 12535602 TI - Aging and temperature effects on DOC and elemental release from a metal contaminated soil. AB - The combined effect of time and temperature on elemental release and speciation from a metal contaminated soil (Master Old Site, MOS) was investigated. The soil was equilibrated at 10, 28, 45, 70 and 90 degrees C for 2 days, 2 weeks, and 2 months in the laboratory. Dissolved organic carbon (DOC), total soluble elements (by ICP), and labile metals (by DPASV) were determined in the filtered (0.22 microm) supernatants. For the samples equilibrated at 90 degrees C, DOC fractions were size fractionated by filtration and centrifugation; a subsample was only centrifuged while another was also filtered through a 0.45 microm filter. Analyses of the supernatants (ICP, DPASV, DOC) were performed on all size fraction subsamples. Dissolved organic carbon (DOC) increased both with temperature and incubation time; however, metal behavior was not as uniform. In general, total soluble metal release (ICP) paralleled the behavior of DOC, increasing with both time and temperature, and confirming the importance of soil organic matter (SOM) in metal retention. Voltammetric analysis (dpasv) of Cu and Zn showed that very little of these metals remains labile in solution due, presumably, to complexation with dissolved organic matter. Labile concentrations of Cd, on the other hand, constituted a significant portion (50%) of total soluble Cd. Copper and Al increased in solution with time (up to 2 months) and temperature up to 70 degrees C; however, at 90 degrees C the soluble concentration declined sharply. The same behavior was observed after equilibration for longer periods of time (550 days) at lower temperatures (23 and 70 degrees C). While concentrations of labile Cu and total soluble Cu and Al increased in the unfiltered samples, the trend remained the same. DPASV analysis showing shifts in labile Cu complexes with temperature and time, together with the results from the unfiltered samples, lead to the hypothesis that Cu was complexing with large polymers that could form at the elevated temperature, and thus be removed from the analyzed solution. It is possible that Cu and Al released by SOM oxidation has re-sorbed or complexed to more recalcitrant organic matter or to mineral phases. Variations in the relative molecular size fractions present within the DOC pool produced by increased time and temperature may influence the element-DOC complexes present in solution and their behavior in soil environments. PMID- 12535603 TI - Effects of airborne volatile organic compounds on plants. AB - Routine measurements of volatile organic compounds (VOCs) in air have shown that average concentrations are very much smaller than those used in laboratory experiments designed to study the effects of VOCs on plants. However, maximum hourly concentrations of some VOCs can be 100 times larger than the average, even in rural air. Experimental studies have rarely extended for longer than a few days, so there is little information on potential long-term effects of exposure to small concentrations. This review considers the available evidence for long term effects, based on laboratory and field data. Previous reviews of the literature from Germany and the USA are cited, prior to an assessment of the effects of individual VOCs. Although hydrocarbons from vehicle exhausts have been implicated in the observed effects on roadside vegetation, the evidence suggests that it is the nitrogen oxides in the exhaust gases that are mostly responsible. There is evidence that aromatic hydrocarbons can be metabolised in plants, although the fate of the metabolites is not known. There is a large literature on the effects of ethylene, because of its role as a plant hormone. Effects have been reported in the field, in response to industrial emissions, and dose response experiments over several weeks in laboratory studies have clearly identified the potential for effects at ambient concentrations. The main responses are morphological (e.g. epinasty), which may be reversible, and on the development of flowers and fruit. Effects on seed production may be positive or negative, depending on the exposure concentration. Chlorinated hydrocarbons have been identified as potentially harmful to vegetation, but only one long-term experiment has studied dose-response relationships. As for ethylene, the most sensitive indication of effect was on seed production, although long-term accumulation of trichloroacetic acid in tissue may also be a problem. There is little evidence of the direct effects of oxygenated hydrocarbons on plants. Plants are a significant emission source of short-chain alcohols, aldehydes and ketones. Peroxyacetyl nitrate (PAN) has a well-documented history as damaging to vegetation. There have been few long-term experimental studies despite the field evidence for damaging effects. Early studies in California have been followed by more recent data from east Asia, but there is still a dearth of information on the potential for effects of PAN and related peroxyacyl nitrates on vegetation typical of regions around tropical and sub-tropical cities where PAN pollution is increasingly important. The lack of long-term measurements, coupled with the available evidence that effects are not linearly related to 'dose' measured as the product of exposure concentration and time, means that the possibility of adverse effects of VOCs on vegetation cannot be safely rejected, particularly in urban and industrial areas. Although reproductive processes (flowering, seed production) appear to be most sensitive, there have been no experimental studies on subsequent seed viability and the consequences at the ecosystem level of changes to plant phenology. The potential for VOC metabolites to accumulate in plant tissue has been demonstrated, but any subsequent effects on herbivores and phytophagous insects have yet to be investigated. PMID- 12535604 TI - Change in maltose- and soluble starch-hydrolyzing activities of chimeric alpha glucosidases of Mucor javanicus and Aspergillus oryzae. AB - The chimeric alpha-glucosidases of Mucor javanicus and Aspergillus oryzae, which has high activity toward not only maltooligosaccharides but also soluble starch and has high activity toward maltooligosaccharides but faint activity toward soluble starch, respectively, were constructed by shuffling the C-terminal regions where low homology is observed between the two enzymes. The chimera genes were expressed in Pichia pastoris to produce and secrete the enzymes that have predicted molecular masses in the culture medium. The two chimeric M. javanicus alpha-glucosidases, of which the N- and C-terminal regions are substituted for those of A. oryzae, respectively, decreased in soluble starch-hydrolyzing activity, however, increased in maltose-hydrolyzing activity by 2.1 and 4.9 times higher than that of the native form of M. javanicus alpha-glucosidase, respectively. The chimeric enzymes changed on the V(max) values for maltose significantly, whereas the K(m) values were similar to that of the native enzyme. PMID- 12535605 TI - Change and stabilization of the amyloid-beta(1-40) secondary structure by fluorocompounds. AB - The misfolding of the amyloid peptide, which is the result of a well-known alpha to-beta transition, causes neurodegenerative disorder. Fluorinated alcohols have been described in the literature as potent solvents which can refold the beta conformation. The present studies demonstrate the effectiveness of differently fluorinated alcohols for the beta-to-alpha refolding process on fibrillar aggregated amyloid beta(1-40). The regenerated helical structure is shown to be maintained in the absence of the fluoroalcohols, a behaviour which was found to contrast with immunoglobulin. We interpret this difference on the basis of the hydrophilic/hydrophobic domains in the amyloid sequence and present some speculations regarding the free-energy levels of the folded states of both proteins. The effect of the -CF(3) group on the observed conformational changes is interpreted as a result of alterations of the hydration shell of the peptides. Moreover, based on the results achieved with fluoroalcohols, we have used novel fluorinated amphiphiles possessing blood-compatibility properties and studied their effect on amyloid beta(1-40). First results point in the direction of a beta-to-alpha transition. Therefore, the use of fluorine groups in the development of new drugs is considered a new possibility requiring further investigation for the prevention of amyloidosis. PMID- 12535606 TI - Expression and characterization of the FXYD ion transport regulators for NMR structural studies in lipid micelles and lipid bilayers. AB - The proteins PLM (phospholemman), CHIF (channel inducing factor), and Mat8 (mammary tumor protein 8 kDa) are members of the FXYD family of ion transport regulatory membrane proteins. Here we describe their cloning and expression in Escherichia coli, and their purification for NMR structural studies in lipid micelles and lipid bilayers. The molecular masses of the purified recombinant FXYD proteins, determined from SDS-PAGE and from MALDI TOF mass spectrometry, reflect monomeric species. The solution NMR and CD spectra in SDS micelles show that they adopt helical conformations. The solid-state NMR spectra in lipid bilayers give the first view of their transmembrane architecture. PMID- 12535607 TI - Multiple sugar binding sites in alpha-glucosidase. AB - Twenty-five analogs of D-glucose were examined as reversible inhibitors of yeast alpha-glucosidase (EC 3.2.1.20). The K(i) values range from 0.38 mM for 6-deoxy-D glucose (quinovose) to 1.0 M for D-lyxose at pH=6.3 (0.1 M NaCl, 25 degrees ). All the monosaccharides and the three disaccharides (maltose, isomaltose and alpha,alpha-trehalose) were found to be linear competitive inhibitors with respect to alpha-p-nitrophenyl glucoside (pNPG) hydrolysis. Multiple inhibition analysis reveals that there are at least three monosaccharide binding sites on the enzyme. One of these can be occupied by glucose [K(i)=1.8(+/-0.1) mM], one by D-galactose [K(i)=164(+/-11) mM] and one by D-mannose [K(i)=120(+/-9) mM]. The pH dependence for glucose binding closely follows that of V/K [pK(a1)=5.55(+/-0.15), pK(a2)=6.79(+/-0.15)], but the binding of mannose does not. Although the glucose subsite can be occupied simultaneously with the mannose or galactose subsites in the enzyme-product complex, no transglucosylation can be detected between pNPG and either mannose or galactose. This suggests that neither of these nonglucose subsites can be occupied in a productive manner in the covalent glucosyl-enzyme intermediate. PMID- 12535608 TI - Biochemical identification of proteasome-associated endonuclease activity in sunflower. AB - Proteasomes have been purified from sunflower hypocotyles. They elute with a molecular mass of 600 kDa from gel filtration columns and two-dimensional gel electrophoresis indicates that the complex contains at least 20 different protein subunits. Peptide microsequencing revealed the presence of four subunits homologous to subunits Beta2, Beta6, Alpha5 and Alpha6 of plant proteasomes. These proteasomes have chymotrypsin-like activity and the highly purified fraction of this complex is associated with an endonuclease activity hydrolyzing Tobacco mosaic virus RNA and Lettuce mosaic virus RNA with a cleavage pattern showing fragments of well-defined size. This is the first evidence of a RNA endonuclease activity associated with plant proteasomes. PMID- 12535609 TI - Structure and function correlations between the rat liver threonine deaminase and aminotransferases. AB - The rat liver threonine deaminase is a cytoplasmic enzyme that catalyses the pyridoxal-phosphate-dependent dehydrative deamination of L-threonine and L-serine to ammonia and alpha-ketobutyrate and pyruvate, respectively, in vivo. During deamination, a molecule of the cofactor is converted to pyridoxamine phosphate. Recently, the ability of this enzyme to accomplish an inverse half-reaction, restoring pyridoxal-phosphate and L-alanine or L-aminobutyrate, respectively, from pyruvate or 2-oxobutyrate, was reported. In order to investigate the molecular mechanisms of this transaminating activity, a molecular model of rat liver threonine deaminase was constructed on the basis of sequence homology with the biosynthetic threonine deaminase of Escherichia coli, the crystal structure of which is known. The model has structural features shared by aminotransferases, suggesting that tertiary structural elements may be responsible for the transaminating activity observed for rat liver threonine deaminase. PMID- 12535610 TI - Disulfide bonds Cys(9)-Cys(57), Cys(34)-Cys(88) and Cys(38)-Cys(90) of the beta subunit of human chorionic gonadotropin are crucial for heterodimer formation with the alpha-subunit: experimental evidence for the conclusions from the crystal structure of hCG. AB - Human chorionic gonadotropin (hCG) is a heterodimeric glycoprotein hormone essential for the establishment and maintenance of pregnancy. The alpha- and beta subunits of hCG are highly cross-linked internally by disulfide bonds that seem to stabilize the tertiary structures required for the noncovalent association of the subunits to generate hormonal activity. This paper describes the results of our studies on the role of the disulfide bonds of hCG-beta in heterodimer formation with the alpha-subunit. Six disulfide peptides incorporating each of the six disulfide bonds of hCG-beta were screened, along with their linear counterparts, for their ability to competitively inhibit the recombination of alpha- and beta-subunits. The disulfide peptides Cys (9-57), Cys (34-88) and Cys (38-90) were found to inhibit the alpha/beta recombination whereas the remaining three disulfide peptides viz. Cys (23-72), Cys (26-110) and Cys (93-100) did not exhibit any inhibition activity. Interestingly, none of the linear peptides could inhibit the alpha/beta recombination. Results clearly demonstrate that the disulfide bonds Cys(9)-Cys(57), Cys(34)-Cys(88) and Cys(38)-Cys(90) of the beta subunit of hCG are crucial for heterodimer formation with the alpha-subunit thus providing experimental confirmation of the conclusions from the crystal structure of the hormone. PMID- 12535611 TI - Water plays a different role on activation thermodynamic parameters of alcoholysis reaction catalyzed by lipase in gaseous and organic media. AB - The effect of water on the alcoholysis of methyl propionate and n-propanol catalyzed by immobilized Candida antarctica lipase B (CALB) has been compared in a continuous solid-gas reactor and in an organic liquid medium. The enthalpic and entropic contributions of water to the Gibbs free energy of activation in the gas phase were different from the ones in the organic phase, the inverse trends being observed for the variation of both DeltaH* and DeltaS* with water activity. Different phenomena were identified for their influence on the thermodynamic parameters. When increasing a(w), the enhanced flexibility of the enzyme was predominant in the gas phase whereas substrate-solvent interactions due to an increased polarity of the solvent affected mainly the thermodynamic parameters in the organic phase. The observed variations of DeltaG* with water activity were in accordance with kinetics results previously obtained in both reaction media. PMID- 12535612 TI - Distinctive interactions in the holoenzyme formation for two isoforms of glutamate decarboxylase. AB - The interactions between glutamate decarboxylase (GAD) and its cofactor pyridoxal phosphate (PLP) play a key role in the regulation of GAD activity. The enzyme has two isoforms, GAD65 and GAD67. A comparison of binding constants, rate constants, and kinetic profiles for the formation of holoenzyme (holoGAD65 and holoGAD67) revealed that the two isoforms interact distinctively with the cofactor. GAD67 exhibits a higher binding constant for PLP binding, making it more difficult to dissociate PLP from holoGAD67 than holoGAD65. Meanwhile, PLP binding occurs at a much slower rate for GAD67 than GAD65, as evidenced by lower rate constants and a slower initial rate of the holoenzyme formation. Job's plots revealed a stoichiometry of 1:1 for PLP binding to GAD65 before and after the saturation level of PLP, while 1:2 for PLP binding to GAD67 prior to the saturation of PLP and 1:1 at the saturation level of PLP. These results suggested that the two binding sites of GAD65 exhibit similar affinities for PLP. In contrast, one binding site of GAD67 exhibits a significantly higher affinity for PLP than the other binding site. Based on these findings, it was proposed that a slower PLP binding to GAD67 than GAD65 and a less ease to dissociate PLP from holoGAD67 than holoGAD65 are important underlying factors. This attributes to GAD67 being more highly saturated by PLP and GAD65 being less saturated by PLP. A larger conformation change constant for GAD67 than GAD65 supported a significant conformational change induced by the initial PLP binding to GAD67, which affects the other binding site affinity of GAD67. The present studies provided valuable insights into distinctive properties between the two isoforms of GAD. PMID- 12535613 TI - A novel DNA polymerase inhibitor and a potent apoptosis inducer: 2-mono-O-acyl-3 O-(alpha-D-sulfoquinovosyl)-glyceride with stearic acid. AB - Sulfo-glycolipids in the class of sulfoquinovosyl diacylglycerol (SQDG) including the stereoisomers are potent inhibitors of DNA polymerase alpha and beta. However, since the alpha-configuration of SQDG with two stearic acids (alpha-SQDG C(18)) can hardly penetrate cells, it has no cytotoxic effect. We tried and succeeded in making a permeable form, sulfoquinovosyl monoacylglycerol with a stearic acid (alpha-SQMG-C(18)) from alpha-SQDG-C(18) by hydrolysis with a pancreatic lipase. alpha-SQMG-C(18) inhibited DNA polymerase activity and was found to be a potent inhibitor of the growth of NUGC-3 cancer cells. alpha-SQMG C(18) arrested the cell cycle at the G1 phase, and subsequently induced severe apoptosis. The arrest was correlated with an increased expression of p53 and cyclin E, indicating that alpha-SQMG-C(18) induced cell death through a p53 dependent apoptotic pathway. PMID- 12535614 TI - Isolation by zinc-affinity chromatography of the histidine-proline-rich glycoprotein molecule associated with rabbit skeletal muscle AMP deaminase. Evidence that the formation of a protein-protein complex between the catalytic subunit and the novel component is critical for the stability of the enzyme. AB - The histidine-proline-rich glycoprotein (HPRG) component of rabbit skeletal muscle AMP deaminase under denaturing and reducing conditions specifically binds to a Zn(2+)-charged affinity column and is only eluted with an EDTA-containing buffer that strips Zn(2+) from the gel. The isolated protein is homogeneous showing an apparent molecular weight (MW) of 95000 and the N-terminal sequence L T-P-T-D-X-K-T-T-K-P-L-A-E-K-A-L-D-L-I, corresponding to that of rabbit plasma HPRG. The incubation with peptide-N-glycosidase F promotes the reduction of the apparent MW of isolated HPRG to 70000, characterizing it as a N-glycosylated protein. The separation from AMP deaminase of an 85-kDa component with a blocked N terminus is observed when the enzyme is applied to the Zn-charged column under nondenaturing conditions. On storage under reducing conditions, this component undergoes an 85- to 95-kDa transition yielding a L-T-P-T-D-X-K-T-T-K-P-L N terminal sequence, suggesting that the shift in the migration on SDS/PAGE as well as the truncation of the protein at its N terminus are promoted by the reduction of a disulfide bond present in freshly isolated HPRG. The separation of HPRG induces a marked reduction in the solubility of AMP deaminase, strongly suggesting a role of HPRG in assuring the molecular integrity of the enzyme. PMID- 12535615 TI - Characterization of short-chain dehydrogenase/reductase homologues of Escherichia coli (YdfG) and Saccharomyces cerevisiae (YMR226C). AB - Short-chain dehydrogenase/reductase homologues from Escherichia coli (YdfG) and Saccharomyces cerevisiae (YMR226C) show high sequence similarity to serine dehydrogenase from Agrobacterium tumefaciens. We cloned each gene encoding YdfG and YMR226C into E. coli JM109 and purified them to homogeneity from the E. coli clones. YdfG and YMR226C consist of four identical subunits with a molecular mass of 27 and 29 kDa, respectively. Both enzymes require NADP(+) as a coenzyme and use L-serine as a substrate. Both enzymes show maximum activity at about pH 8.5 for the oxidation of L-serine. They also catalyze the oxidation of D-serine, L allo-threonine, D-threonine, 3-hydroxyisobutyrate, and 3-hydroxybutyrate. The k(cat)/K(m) values of YdfG for L-serine, D-serine, L-allo-threonine, D-threonine, L-3-hydroxyisobutyrate, and D-3-hydroxyisobutyrate are 105, 29, 199, 109, 67, and 62 M(-1) s(-1), and those of YMR226C are 116, 110, 14600, 7540, 558, and 151 M( 1) s(-1), respectively. Thus, YdfG and YMR226C are NADP(+)-dependent dehydrogenases acting on 3-hydroxy acids with a three- or four-carbon chain, and L-allo-threonine is the best substrate for both enzymes. PMID- 12535616 TI - Secretory proprotein convertases PACE4 and PC6A are heparin-binding proteins which are localized in the extracellular matrix. Potential role of PACE4 in the activation of proproteins in the extracellular matrix. AB - PACE4, PC6 and furin are potent subtilisin-like proprotein convertases (SPCs) which are responsible for the activation of transforming growth factor-beta (TGFbeta)-related factors such as bone morphogenetic proteins. Heparan sulfate proteoglycan within the extracellular matrix (ECM) is known to regulate the biological activity of various differentiation factors including TGFbeta-related molecules. PACE4 binds tightly to heparin and its heparin-binding region was found to be a cationic stretch of amino acids between residues 743 and 760. Furthermore, PACE4 was detected in the extracellular material fraction of the HEK293 cells, defined as the material remaining on the culture plate following the removal of the cells from the plate. PACE4 bound to the extracellular fraction was selectively dislodged by heparin into the culture medium. Heparin has no inhibitory activity against PACE4. Similarly, PC6A is also able to bind to heparin, whereas soluble furin does not. In human placenta, PACE4 is mainly present in syncytiotrophoblasts and can be released by heparin. These results suggest that PACE4 and PC6 are unique SPC family proteases that anchor heparan sulfate proteoglycans at the ECM. The interaction between PACE4 and heparan sulfate proteoglycans might play an important role in the delicate spatiotemporal regulation of TGFbeta-related factors' biological activity. PMID- 12535617 TI - Contributions of individual residues in the N-terminal region of cystatin B (stefin B) to inhibition of cysteine proteinases. AB - The importance of individual residues in the N-terminal region of cystatin B for proteinase inhibition was elucidated by measurements of the affinity and kinetics of binding of N-terminally truncated, recombinant variants of the bovine inhibitor to cysteine proteinases. Removal of Met-1 caused an 8- to 10-fold lower affinity for papain and cathepsin B, decreased the affinity also for cathepsin L but only minimally affected cathepsin H affinity. Additional truncation of Met-2 further weakened the binding to papain and cathepsin B by 40-70-fold, whereas the affinity for cathepsins L and H was essentially unaffected. Removal of Cys-3 had the most drastic effects on the interactions, resulting in a further affinity decrease of approximately 1500-fold for papain, approximately 700-fold for cathepsin L and approximately 15-fold for cathepsin H; the binding to cathepsin B could not be assessed. The binding kinetics could only be evaluated for papain and cathepsin H and showed that the reduced affinities for these enzymes were predominantly due to increased dissociation rate constants. These results demonstrate that the N-terminal region of cystatin B contributes appreciably to proteinase inhibition, in contrast to previous proposals. It is responsible for 12-40% of the total binding energy of the inhibitor to the proteinases investigated, being of least importance for cathepsin H binding. Cys-3 is the most important residue of the N-terminal region for inhibition of papain, cathepsin L and cathepsin H, the role of the other residues of this region varying with the target proteinase. PMID- 12535618 TI - Crystallization and preliminary X-ray crystallographic analysis of plant ferritin from Glycine max. AB - The iron storage protein ferritin from soybean (Glycine max) was expressed in E. coli and crystallized using the hanging drop vapor diffusion method with sodium tartrate as the precipitant. The crystals belong to the tetragonal I4(1)22 space group, with unit cell parameters a=b=324.0, c=182.7 A. The diffraction data were collected up to a resolution of 3.0 A with a multi-wire area detector. PMID- 12535619 TI - In vitro action of leptin on FSH and LH production in rainbow trout (Onchorynchus mykiss) at different stages of the sexual cycle. AB - The short-term effect of recombinant human leptin (rhleptin) on FSH and LH production (release+intracellular content) was studied in vitro using pituitary cells from male and female rainbow trout during the first gametogenesis cycle. In our rearing conditions, we found a direct action of rhleptin at the pituitary level, which depends on the sexual stage of the fish. No effect of rhleptin on FSH or LH release and cellular content could be detected in immature fish and post-ovulatory females. However, throughout the process of spermatogenesis and ovogenesis, high concentrations (0.5 and 1 x 10(-6)M) of rhleptin stimulated FSH and LH release, without observable action on intracellular content of gonadotropins. A relatively constant response to rhleptin for FSH was observed throughout gonad maturation, while LH response tended to be higher at the first stages of gametogenesis (beginning of spermatogenesis and endogenous vitellogenesis). Preliminary results on the potential interaction of rhleptin and salmon GnRH (sGnRH) suggest a possible synergistic effect of high concentration of rhleptin (10(-6)M) and sGnRH only at restricted phases of gonadal development when the gametogenetic process was already fully started (full spermatogenesis and early vitellogenesis). The direct action of leptin on FSH and LH release, evident only when gametogenesis had already started suggests that leptin is not the unique signal for the activation of the gonadotropic axis but requires a combined action with other promoting factors. PMID- 12535620 TI - Hypothalamic indolamines during embryonic development and effects of steroid exposure. AB - The serotonin system has been implicated in the modulation of endocrine and behavioral components of reproduction. In this study, we examined endogenous hypothalamic indolamines during sexual differentiation and long-term effects of exogenous steroids during this time. In Experiment 1, Japanese quail were studied during the last half of embryonic development and early post-hatch. Samples were taken at embryonic day 10 (E10), E12, E14, E16, hatch (day 0), and days 3 and 5, post-hatch. Hypothalamic indolamines, including serotonin (5-HT) and its metabolite, 5-hydroxy indole acetic acid (5-HIAA) were measured by HPLC-EC detection. Females had relatively higher hypothalamic 5-HT at E14 than males, with both sexes showing increasing levels thereafter. By day 5, post-hatch, hypothalamic 5-HT content was higher in males than in females. When turnover was estimated by comparing relative concentrations of 5-HT to 5-HIAA, males were significantly higher at E12 and E14 than females. These data suggest that there are stage specific changes in the serotonin system, as well as sexually dimorphic patterns in the ontogeny and activity of this system. In Experiment 2, we investigated the effects of embryonic steroid hormone treatment on the serotonin system and on male sexual behavior. Birds were treated with either estradiol benzoate (EB), testosterone propionate (TP) or sesame oil (vehicle control) at selected embryonic days (E10, E12, E14, E16, 0, D3, and D5). At 4 weeks post hatch, birds were transferred to short photoperiod (16D:8L) for 3 weeks to prevent photostimulated reproductive development. At 7 weeks of age, males were implanted with a 20mm silastic capsule filled with testosterone and sexual behavior was tested 1 week later. Brains were collected from both males and females, and preoptic area (POA) indolamines were measured. Steroid treatment at E10 or E12 resulted in the loss of male sexual behavior. Moreover, males treated with EB or TP on E12 also had increased POA 5-HT content as adults, compared to control males. Females treated with EB on either E10 or E 12 also had higher POA 5-HT content than control or TP treated females. These data provide evidence for sexual dimorphism in the hypothalamic 5-HT system at specific stages during embryonic development. Moreover, males were sensitive to exogenous EB and TP on E12, whereas females appeared to be affected by EB only and appeared to be sensitive to steroid effects over a longer period of time in development. Moreover, exogenous steroids at E12 in males also correlated with impaired sexual behavioral. These data suggest that long-term effects of embryonic steroid exposure may be mediated in part through effects on the serotonin neurotransmitter system. PMID- 12535621 TI - Localization of gonadotropin-releasing hormone in the central nervous system and a peripheral chemosensory organ of Aplysia californica. AB - Gonadotropin-releasing hormone (GnRH) is a neurohormone crucial for the regulation of reproductive and neural functions in vertebrates. Recent discoveries of GnRH immunoreactivity (IR) in a number of invertebrates raised the possibility that GnRH may be an ancient molecule that had arisen before the emergence of Phylum Chordata. We previously demonstrated the presence of a GnRH IR similar to the mammalian (m) and tunicate I (tI) forms of GnRH in the hemolymph and ovotestis of an opisthobranch mollusk, Aplysia californica; however, the presence of GnRH in the central nervous system (CNS) of A. californica could not be detected with the available antisera against various forms of chordate GnRH. In the present study, we performed immunohistochemistry (IHC) to localize the presence of GnRH in the CNS and a peripheral chemosensory organ, the osphradium, of A. californica. A newly generated antiserum against tI GnRH revealed the strong expression of GnRH IR in neurons of all CNS ganglia. A notable asymmetry in immunostaining was detected in the left and right abdominal hemiganglia. The CNS is rich in tI-GnRH immunoreactive neurons but lacks mGnRH IR, whereas the osphradium contains abundant mGnRH immunoreactive neurons but lacks tI-GnRH IR. The extract of CNS failed to stimulate the release of LH from mouse pituitary, demonstrating that the A. californica GnRH IR is structurally different from what is required to bind and activate mammalian GnRH receptor. Together, these results indicate the presence of at least two distinct GnRH systems in A. californica. The presence of GnRH in the osphradium is consistent with the long-standing anatomical relationship between GnRH and the chemosensory system observed in vertebrates. PMID- 12535622 TI - Brain tyrosine hydroxylase in the catfish Heteropneustes fossilis: annual and circadian variations, and sex and regional differences in enzyme activity and some kinetic properties. AB - Dynamics of tyrosine hydroxylase (TH) was studied in various brain regions and pituitary in relation to annual/seasonal and circadian variations in either sex of the catfish Heteropneustes fossilis. alpha-Methylparatyrosine inhibited TH activity significantly in a time-(in vivo study) and concentration-(in vitro study) dependent manner. The inhibition was higher in resting phase than preparatory phase and in females than males (time-course study). The apparent kinetic constant of inhibition (K(i)) varied both seasonally and regionally. The values were higher in resting phase and in the forebrain regions (telencephalon and hypothalamus) than medulla oblongata. No significant sex difference was noticed in the K(i) values in the same season. TH activity showed significant annual/seasonal variations with telencephalon and hypothalamus showing higher activity than medulla oblongata. The females showed significantly higher enzyme activity than males with a distinct activity peak in June (prespawning phase). Such an activity peak was apparently absent in males. TH activity showed significant circadian/diurnal variations, the highest activity was noticed at 12 h and the lowest at 24 h. The apparent Michaelis-Menten constant (K(m)) values (hypothalamic TH) for substrate (L-tyrosine) and cofactor (DMPH(4)) showed seasonal variations with the highest values in the resting phase, decreasing through preparatory and prespawning phases, to the lowest values in spawning phase. V(max) was the lowest in the resting phase and highest in the spawning phase. TH activity was low in the pituitary and could be detected in pooled samples from March (preparatory phase) to July (spawning phase). Activity showed significant variations, which could be correlated with the gonadosomatic index. The results show that TH activity could be positively correlated with the annual reproductive cycle. The enzyme activity could be also correlated with seasonal, sex, and regional variations in the apparent K(m) and V(max) values suggesting apparent differences in the affinity of the enzyme towards substrate and cofactor. PMID- 12535623 TI - Dopamine infusion into the third ventricle increases gene expression of hypothalamic vasoactive intestinal peptide and pituitary prolactin and luteinizing hormone beta subunit in the turkey. AB - Turkey prolactin (PRL) secretion is controlled by vasoactive intestinal peptide (VIP) neurons residing in the infundibular nuclear complex (INF) of the hypothalamus. The VIPergic activity is modulated by dopamine (DA) via stimulatory D(1) DA receptors. DA (10 nmol/min for 40 min) was infused into the third ventricle of laying turkey hens to study its effect on circulating PRL, hypothalamic VIP and pituitary PRL and LHbeta subunit mRNA levels. Plasma PRL was significantly elevated after 20 min of DA infusion and remained elevated 30 min after cessation of infusion. Hypothalamic VIP mRNA content was significantly greater in the INF of DA-infused birds than it was in the INF of vehicle-infused control birds. No increase in VIP mRNA due to DA infusion was noted in the preoptic area. Pituitary PRL and LHbeta subunit mRNAs were increased in DA infused hens as compared to vehicle-infused controls but the rate of increase was more in PRL than LHbeta subunit. This study demonstrates that exogenous DA activates hypothalamic VIP gene expression and this increased expression is limited exclusively to the avian INF. The increased VIP mRNA in the INF is correlated with increased levels of circulating PRL and PRL and LHbeta mRNAs in the anterior pituitary. PMID- 12535624 TI - Effects of season, sex, and sample collection on concentrations of fecal cortisol metabolites in red deer (Cervus elaphus). AB - Seasonal variation, sex differences, and invasive sample collection may confound glucocorticoid measures as indices of stress. We investigated the effects of sex and season on glucocorticoid production on a non-invasive basis by measuring concentrations of cortisol metabolites in feces of undisturbed red deer (Cervus elaphus). Although feces can be collected easily, assignment to individuals is difficult. Anonymous fecal samples may cause overrepresentation of particular individuals thus introducing a source of error when estimating mean hormone levels within a population. We therefore examined the effects of collecting anonymous fecal samples on mean fecal cortisol metabolite levels. Neither sex nor sample collection mode significantly affected mean fecal cortisol metabolite concentrations in the studied population of red deer. Fecal glucocorticoid excretion varied seasonally with a peak during December and January. Out of several potential predictor variables investigated, minimum ambient temperature and snow proved to be the only factors exerting a significant effect on fecal glucocorticoid excretion. We suggest that high winter glucocorticoid levels may act via catabolic function during adaptation of deer to cold winter month when resources are limited. PMID- 12535625 TI - Seasonal changes of responses to gonadotropin-releasing hormone analog in expression of growth hormone/prolactin/somatolactin genes in the pituitary of masu salmon. AB - Gonadotropin-releasing hormone (GnRH) is considered to stimulate secretion of growth hormone (GH), prolactin (PRL), and somatolactin (SL) at particular stages of growth and sexual maturation in teleost fishes. We therefore examined seasonal variation in the pituitary levels of GH/PRL/SL mRNAs, and tried to clarify seasonal changes of responses to GnRH in expression of GH/PRL/SL genes, in the pituitaries of growing and maturing masu salmon (Oncorhynchus masou). Pituitary samples were monthly collected one week after implantation with GnRH analog (GnRHa). The levels of mRNAs encoding GH, PRL, and SL precursors in single pituitaries were determined by a real-time polymerase chain reaction method. The fork lengths and body weights of control and GnRHa-implanted fish of both sexes gradually increased and peaked out in September of 2-year-old (2+) when fish spawned. GnRHa implantation did not stimulate somatic growth, nor elevate gonadosomatic index (GSI) of 1+ and 2+ males, whereas it significantly increased GSI of 2+ females in late August to early September. The GnRHa-implanted 1+ males had higher levels of GH and PRL mRNAs in July, and SL mRNA from June to August than the control males. The levels of GH, PRL, and SL mRNAs in the control and GnRHa-implanted 1+ females, however, did not show any significant changes. Afterward, the PRL mRNA levels elevated in the control 2+ fish of both sexes in spring. GnRHa elevated the GH mRNA levels in both males and females in 2+ winter, and the PRL mRNA levels in females in early spring. Regardless of sex and GnRHa implantation, the SL mRNA levels increased during sexual maturation. In growing and maturing masu salmon, expression of genes encoding GH, PRL, and SL in the pituitary is thus sensitive to GnRH in particular seasons probably in relation to physiological roles of the hormones. PMID- 12535626 TI - Comparative urinary androstanes in the great apes. AB - Urinary androstanes from seven species of male great apes (human, bonobo, chimpanzee, lowland gorilla, mountain gorilla, Bornean orangutan, and Sumatran orangutan) were separated by HPLC and detected by RIA using two testosterone antibodies. All animals examined showed the presence of testosterone and six additional immunoreactive peaks. Although testosterone was the dominant peak (85%) in human urine, its proportion in urine was much less in the other apes, ranging from a high of 59% in the bonobo and chimpanzee to a low of 24% in the mountain gorilla. Urinary androstanes were also directly visualized using nano spray mass spectrometry (nanoESI-MS). Although the RIA can qualitatively produce a strong signal for testosterone in unchromatographed urine, it is quantitatively present only as a trace metabolite, as demonstrated by nanoESI-MS. The combination of the two techniques showed large differences in androstane metabolism between the seven species. A previously undescribed testosterone metabolite (tentatively identified as either delta1- or delta6-testosterone sulfate) was present in significant proportions in all of the non-human apes examined. We conclude that in the great apes, testosterone is only a trace metabolite in urine, and as a consequence, its measurement may not produce results that parallel the levels of serum testosterone. The RIA measurement of urinary testosterone in part records additional androstane metabolites, which vary even between closely related genera, making the results neither equivalent with nor comparable to different species. PMID- 12535627 TI - Cold-induced enhancement of avian uncoupling protein expression, heat production, and triiodothyronine concentrations in broiler chicks. AB - The relationships among avian uncoupling protein (avUCP) mRNA expression, heat production, and thyroid hormone metabolism were investigated in 7-14-day-old broiler chicks (Gallus gallus) exposed to a low temperature (cold-exposed chicks, CE) or a thermoneutral temperature (TN). After 7 days of exposure, CE chicks exhibited higher heat production (+83%, P<0.01), avUCP mRNA expression (+20%, P<0.01), and circulating triiodothyronine (T(3)) levels (+104%, P=0.07) for non statistically different body weights and feed intake between 3 and 7 days of exposure as compared to TN chicks. Plasma thyroxine (T(4)) concentration was clearly decreased in CE chicks (-33%, P=0.06). The lower hepatic inner-ring deiodination activity (-47%) and the higher renal outer-ring deiodination activity (+75%) measured in CE compared to TN chicks could partly account for their higher plasma T(3) concentrations. This study describes for the first time the induction of avUCP mRNA expression by low temperature in chickens, as it has been previously shown in ducklings, and supports the possible involvement of avUCP in avian thermogenesis. PMID- 12535628 TI - Hormonal changes associated with the transition between nursing and natural fasting in northern elephant seals (Mirounga angustirostris). AB - To better interpret previously described hormonal changes observed during the natural postweaning fast (2-3 months) endured by pups of the northern elephant seal (Mirounga angustirostris), we compared plasma cortisol, thyroid hormones, and leptin in pups (n=5) measured during nursing and fasting periods. Blood samples were taken at four times; early (9 days postpartum) and late (18-22 days postpartum) nursing, and early (second week postweaning) and late (eighth week postweaning) fasting. Plasma cortisol increased 39% between early and late nursing and almost 4-fold by late fasting. After the early nursing period, cortisol and body mass were negatively correlated (y=28.3-0.19 x; R=0.569; p=0.027). Total thyroxine (tT(4)), free T(4) (fT(4)), total triiodothyronine (tT3) and reverse T(3) (rT(3)) were greatest at early nursing and reduced by late nursing and remained so throughout the fast, with the exception of tT(4), which increased between late nursing (17.7+/-2.1 ng mL(-1)) and late fasting (30.1+/ 2.8 ng mL(-1)) periods. Leptin remained unaltered among the four sampling periods and was not correlated with body mass. Pups appear to exhibit a shift in the relationship between cortisol and body mass suggesting a potential role for cortisol in the regulation of body fat. The higher concentrations of tT(3) and tT(4) during early nursing may reflect enhanced growth and development during this period, however the increase late in fasting is likely physiologically insignificant and an artifact of reduced metabolic clearance of these hormones. Transition of the pups from nursing to fasting states is characterized by a striking lack of change in cortisol, thyroid hormones, and leptin suggesting that any metabolic alterations associated with this transition may occur independent of these hormones. PMID- 12535629 TI - The liver of Fundulus heteroclitus expresses deiodinase type 1 mRNA. AB - The presence of a type 1 deiodinase (D1) in the liver of teleosts has been a controversial issue. Recently we characterized the deiodinase activity in rainbow trout and killifish liver and found that the liver of both species co-expresses the two enzymes (D1 and D2) that catalyze the outer ring-deiodinating pathway. We here report the cloning and characterization of an mRNA from the liver of the killifish Fundulus heteroclitus that encodes a D1 (FhD1). The cDNA amplified by RT-PCR from F. heteroclitus liver is 1314 nt long and encodes a protein of 248 aa. It contains a TGA codon in its open reading frame and a selenocysteine insertion sequence in its 3(') untranslated region, consistent with the structure of a selenoenzyme mRNA. The deduced peptide sequence is 73% identical to that encoded by the tilapia D1 cDNA cloned from kidney and 46% identical to the D1s reported in other vertebrates. Northern blot analysis shows that FhD1 mRNA is expressed in F. heteroclitus liver, consistent with prior biochemical evidence for hepatic D1 activity. Furthermore, heterologous expression of the FhD1 cDNA resulted in a protein with properties similar to the D1-like activity in F. heteroclitus liver. The cloned enzyme, like the native species, is relatively insensitive to inhibition by PTU, but mutation of Ser-159 in FhD1 to the Pro residue found in D2 and D3 isoforms increased the sensitivity to PTU. Our results show that, under basal conditions, killifish liver indeed expresses a D1 enzyme that is homologous to mammalian D1s, establishing this as a useful model in which to study the regulation of D1 and D2 concurrently. PMID- 12535630 TI - Isolation, synthesis, and biological activity of flounder [Asn1,Ile5,Thr9] angiotensin I. AB - A novel angiotensin I (ANG I) has been isolated from incubates of plasma and kidney extracts of the flounder, Platichthys flesus, using ion-exchange, gel permeation, and reverse-phase high performance liquid chromatography (HPLC). Its sequence was determined as H-Asn-Arg-Val-Tyr-Ile-His-Pro-Phe-Thr-Leu-OH by sequence analysis and mass spectrometry. No vasopressor activity was detected at the elution position of [Asp(1)] ANG I in ion-exchange HPLC. The sequence was confirmed by identity of the elution position with the synthetic peptide in two different HPLC systems. When compared with ANG I isolated from other teleost fish, flounder ANG I uniquely has an isoleucine at position 5 rather than valine. Injection of angiotensin II (ANG II) into chronically cannulated flounder resulted in a dose-dependent pressor response, native [Asn(1),Ile(5)] ANG II, was found to elicit pressor responses comparable with those seen when teleost [Asn(1),Val(5)] ANG II and human [Asp(1),Ile(5)] ANG II were injected into flounder over the dose range 0.02-1.00 nmol/kg(-1). Plasma concentrations of the neurohypophysial peptide AVT were measured in chronically cannulated flounder following the injection of ANG II to examine the effect of ANG II on circulating AVT concentration. The injection of [Asn(1),Ile(5)] ANG II (1 nmolkg(-1)) or [Asp(1),Ile(5)] ANG II (2.5 nmolkg(-1)) resulted in a significant fall in the circulating levels of AVT suggesting that ANG II either directly or indirectly negatively influences AVT secretion. PMID- 12535631 TI - Structural and functional analysis of 3' untranslated region of mouse Period1 mRNA. AB - In order to investigate the post-transcriptional regulation of Period1 (Per1), the 3(')-untranslated region (3(')UTR) of mouse Per1 (mPer1) mRNA was characterized. In addition to high similarity between human and mouse Per1 3(')UTRs, AU-rich element and differentiation control element were found in both species. Transient transfection assays using LUC::mPer1 3(')UTR fusion genes revealed that the mPer1 3(')UTR repressed its own expression in a post transcriptional manner. The region critical for this translational down regulation was confined to nucleotide positions 322-517. These results suggest that the mPer1 3(')UTR could be involved in the generation of time lag between the transcriptional and translational products of mPer1 in the suprachiasmatic nucleus. PMID- 12535632 TI - Inhibition of human intestinal brush border membrane vesicle Na+-dependent phosphate uptake by phosphophloretin derivatives. AB - Hyperphosphatemia and II(o) hyperparathyroidism are common and severe complications of chronic renal failure. Reduced dietary phosphorus has been shown to be an effective treatment in reducing serum phosphate and serum PTH. 2(') Phosphophloretin inhibited small intestine apical membrane Na(+)/phosphate cotransport and reduced serum phosphate in adult rats. 2(')-PP and phosphoesters of phloretin were tested for inhibition of human small intestine brush border membrane alkaline phosphatase activity and for inhibition of Na(+)-dependent phosphate uptake. The IC(50)'s for inhibition of alkaline phosphatase suggested an order of inhibitory potency of 4-PP > phloretin > 4(')-PP > 2(')-PP. Inhibition of Na(+)-dependent phosphate uptake followed the sequence 2(') PPz.Gt;4(')-PP > 4-PP > phloretin. These results are consistent with 2(')-PP being a specific inhibitor of human intestinal brush border membrane Na(+)/phosphate cotransport. PMID- 12535633 TI - Mastoparan induces Ca2+-independent cortical granule exocytosis in sea urchin eggs. AB - In most species, cortical granule exocytosis is characteristic of egg activation by sperm. It is a Ca(2+)-mediated event which results in elevation of the vitelline coat to block permanently the polyspermy at fertilization. We examined the effect of mastoparan, an activator of G-proteins, on the sea urchin egg activation. Mastoparan was able to induce, in a concentration-dependent manner, the egg cortical granule exocytosis; mastoparan-17, an inactive analogue of mastoparan, had no effect. Mastoparan, but not sperm, induced cortical granule exocytosis in eggs preloaded with BAPTA, a Ca(2+) chelator. In isolated egg cortical lawns, which are vitelline layers and membrane fragments with endogenously docked cortical granules, mastoparan induced cortical granule fusion in a Ca(2+)-independent manner. By contrast, mastoparan-17 did not trigger fusion. We conclude that in sea urchin eggs mastoparan stimulates exocytosis at a Ca(2+)-independent late site of the signaling pathway that culminates in cortical granule discharge. PMID- 12535634 TI - TGF-beta1 induces the expression of fast inactivating K+ (I(A)) channels in rat vascular myofibroblasts. AB - It is well established that transforming growth factor-beta1 (TGF-beta1) can induce the transformation of fibroblasts to myofibroblasts. The molecular mechanisms of the phenotypic change remain unknown. The effect of TGF-beta1 on the expression of K(+) channels in cultured rat vascular fibroblasts was investigated by using the patch-clamp technique and quantitative RT-PCR. In fibroblasts, the only voltage-dependent outward K(+) current that can be electrophysiologically detected is non-inactivating. In myofibroblasts, induced by the treatment of fibroblasts with TGF-beta1, we report the emergence of an additional transient outward K(+) current The TGF-beta1-induced outward current is inhibited by 4-aminopyridine. K(V2.1), the transcript for a non-inactivating potassium channel gene, was detected by quantitative RT-PCT in both cultured fibroblasts and myofibroblasts. In contrast, the transcript of the transient I(A) gene, K(V4.1), can be detected only in myofibroblasts. The results suggest that TGF-beta1-induced phenotypic transformation of vascular fibroblasts to myofibroblasts is accompanied by the induction of I(A) channels. PMID- 12535635 TI - A real-time analysis of QacR-regulated multidrug resistance in Staphylococcus aureus. AB - Here we describe the construction and characterization of a biosensing reporter where luxCDABE genes from Photorhabdus luminescens, engineered for expression in Gram-positive organisms, are under the transcriptional control of QacR repressor from Staphylococcus aureus. In non-pathogenic S. aureus model system we analyzed the activity of the regulatory region acting as multidrug-resistance mediator in wild type strains. The use of full-length bacterial luciferase and the measurement of real-time light emission from intact, living cells make the present system suitable to follow the short term activity of different inducers. Among the tested molecules, tetracyclines showed a peculiar behavior by giving very high induction in a fashion seemingly more related to a general stress condition of the culture than to the direct binding and displacement of QacR from its operator. Temperature shocks confirmed that active transcription from qacA promoter is started in response to unspecific conditions where cell growth is strongly inhibited. PMID- 12535636 TI - Alpha-hairpin stability and folding of transmembrane segments. AB - Molecular Dynamics (MD) simulations at low dielectric constant have been carried out for peptides matching the double spanning segments of transmembrane proteins. Different folding dynamics have been observed. The peptides folded into the stable helix-turn-helix conformation-alpha-hairpin-with antiparallel-oriented strands or unstable alpha-hairpin conformation that unfolded later into the straight helical structure. The peptide having flexible residues in the TM helices often misfolded into a tangled structure that can be avoided by restricting the flexibility of these residues. General conclusions can be drawn from the observed folding dynamics. The stability and folding of some double spanning transmembrane fragments are self-assembling. The following and/or neighboring peptide chains of the protein may support the stability of the hairpin structure of other fragments. The stability of the TM helices containing flexible residues could be maintained due to contacts with neighboring TM segments. PMID- 12535637 TI - Human ADAM33: protein maturation and localization. AB - ADAM33 (a disintegrin and metalloprotease) was recently found to be a novel asthma susceptibility gene. Domain-specific antibodies were used to study its expression and processing. When the pro-domain and catalytic domain were expressed by a stable-transfected cell line, the pro-domain was removed by cleavage within a putative furin cleavage site. The catalytic domain was active in an alpha(2)-macroglobulin complex formation assay and mutation of the catalytic site glutamic acid (E346A) eliminated activity. In transient transfections using the full-length protein, a pro-form and mature form were detectable and alternate glycosylation was demonstrated at sites within the catalytic domain. ADAM33 was detected on the cell surface, with the majority of protein detected intracellularly. The E346A mutation had no significant effect on protein processing. Endogenous ADAM33 was detected in bronchus tissue, bronchial smooth muscle cells, and MRC-5 fibroblasts, consistent with a role in the pathophysiology of asthma. PMID- 12535638 TI - BetaB2-crystallin undergoes extensive truncation during aging in human lenses. AB - Based on the present literature, it is unclear whether betaB2-crystallin undergoes age-related truncation in human lenses. To answer this question, the purpose of this study was to determine in vivo truncation of betaB2-crystallin in human lenses during aging by examining its fragments in the beta(H)-crystallin fraction. The WS-protein fraction was isolated from lenses of desired ages and separated by a size-exclusion Agarose A 1.5m column to recover alpha-, beta(H)-, beta(L)-, and gamma-crystallin fractions. The beta(H)-crystallin fractions, isolated from lenses of 24- and 70-year-old donors, were utilized for two dimensional (2D) gel electrophoresis (isoelectric focusing in the first dimension followed by SDS-PAGE in the second dimension). The partial N-terminal sequences of the desired fragments (Molecular weights [M(r)]<18-19kDa) from a 2D-gel of WS proteins from lenses of a 70-year-old donor were determined. More than 37 crystallin fragments with M(r) between 4 and 19kDa were observed on a 2D-gel. Nine fragments in beta(H)-crystallin fraction were from betaB2-crystallin but additional single fragments of alphaA-, gammas-, betaA4, and of either gammaB-, gammaC- or gammaD-crystallins were also observed. Seven cleavage sites in the betaB2-crystallin were identified, which included two sites at Q(7)-A(8) and A(8) G(9) bonds in the N-terminal extension, two sites at E(46)-K(47) and G(49)-S(50) bonds in the motif 1, one site at S(94) -S(95) in the motif 2, and two sites at N(115)-F(116) and Q(135)-Y(136) in motif 3. No fragments with cleavage in the motif 4 and C-terminal extension of betaB2-crystallin were seen. Apparently, three betaB2-crystallin fragments with only N-terminal cleavage and five with both N- and C-terminal cleavages were observed. Additional fragments with cleavage sites at Q(54)-Y(55) in alphaA-crystallin, at E(112)-N(113) in betaA4 crystallin, at G(4)-T(5) in gammas-crystallin, at M(69)-G(70) in either gammaB-, gammaC- or gammaD-crystallins (three have identical sequences at the cleaved bond), and at G(1)-K(2) in gammaB or gammaC (both have identical sequences at the cleavage site) were observed.Conclusions. The results showed that betaB2 crystallin undergoes age-related truncation producing fragments with M(r) between 4 and 19kDa that existed in the beta(H)-crystallin oligomer. The beta(H) crystallin fraction also contained single fragments of alpha-, betaA4-, gammas-, and other gamma-crystallins. PMID- 12535639 TI - Activation of PPARgamma increases PTEN expression in pancreatic cancer cells. AB - The PI3K pathway contributes to the invasive properties and apoptosis resistance that epitomize pancreatic cancers. PPARgamma is a ligand-activated transcription factor with anti-inflammatory and anti-tumor effects; the mechanisms of tumor suppression are unknown. The purpose of this study was to examine whether activation of PPARgamma can increase the expression of the tumor suppressor PTEN and inhibit PI3K activity. AsPC-1 human pancreatic cancer cells, transfected with a PPRE-luciferase construct, demonstrated increased luminescence following treatment with PPARgamma ligands, indicating the presence of functional PPARgamma protein. The selective PPARgamma ligand rosiglitazone increased PTEN expression in AsPC-1 cells; concurrent treatment with GW9662, which inhibits PPARgamma activation, prevented the increase in PTEN protein levels. Levels of phosphorylated Akt decreased as PTEN levels increased, indicating inhibition of PI3K activity. Taken together, our results suggest that activation of PPARgamma may represent a novel approach for the treatment of pancreatic cancer by increasing PTEN levels and inhibiting PI3K activity. PMID- 12535640 TI - Intracellular delivery of HSP70 using HIV-1 Tat protein transduction domain. AB - Heat shock protein 70 (HSP70) is an intracellular stress protein that confers cytoprotection to a variety of cellular stressors. Several lines of evidence have suggested that augmentation of the heat shock response by increasing the expression of HSP70 represents a potential therapeutic strategy for the treatment of critically ill patients. The Tat protein of human immunodeficiency virus 1 (HIV-1) has been used previously to deliver functional cargo proteins intracellularly when added exogenously to cultured cells. We generated a Tat HSP70 fusion protein using recombinant methods and treated HSF -/- cells with either Tat-HSP70 or recombinant HSP70 prior to exposure to hyperoxia or lethal heat shock. We showed that biologically active, exogenous HSP70 can be delivered into cells using the HIV-1 Tat protein, and that the Tat-mediated delivery of HSP70 confers cytoprotection against thermal stress and hyperoxia and may represent a novel approach to augmenting intracellular HSP70 levels. PMID- 12535641 TI - Generation of affinity matured scFv antibodies against mouse neural cell adhesion molecule L1 by phage display. AB - The recognition molecule L1 plays important functional roles in the nervous system and in non-neural tissues. Since antibodies to L1 are of prime importance to study its functional properties, we have generated affinity matured human single chain variable fragment (scFv) antibodies against mouse L1 by introducing random mutations in the complementarity determining regions (CDRs) of a previously isolated scFv antibody heavy chain (CDR1 and CDR2) and light chain (CDR3). After biopanning the mutant library, a clone (5F7) that gave the strongest ELISA signal was expressed, purified, and characterized. The dissociation constant of 5F7 (2.86 x 10(-8)M) was decreased 60-fold compared to the wild type clone G6 (1.72 x 10(-6)M). 5F7 detected L1 by Western blot analysis in mouse brain homogenates and recognized L1 in L1 transfected cells and cryosections from mouse retina and optic nerve by immunofluorescence. Bivalent 5F7 scFv antibody (5F7-Cys) was also generated and showed a dissociation constant of 5.22 x 10(-9)M that is 5.5-fold lower than that of monomeric 5F7 antibody. The bivalent affinity matured L1 scFv antibody thus showed stronger binding by a factor of 310 compared to the wild type clone. This antibody should be useful in various biological assays. PMID- 12535642 TI - A novel zinc finger protein, ZZaPK, interacts with ZAK and stimulates the ZAK expressing cells re-entering the cell cycle. AB - ZAK has been implicated in cell cycle arrest regulation through its function on decreasing cyclin E expression. To explore the mechanistic basis for this regulation, the yeast two-hybrid system was used with a novel Kruppel-type C2H2 zinc finger member cloned. This cloned cDNA encodes a novel protein with Kruppel type zinc fingers designed as ZZaPK (zinc finger and ZAK associated protein with KRAB domain) and is widely expressed. ZZaPK, when it is expressed in cells, is growth promoted and might lead to increasing E2F expression and induce cyclin E/CDK2 activity, which counteracts the ZAK function. The model proposed here is that ZAK might play a role as an upstream signal to suppress the ZZaPK function and decrease E2F expression. PMID- 12535643 TI - Calcineurin-mediated pathway involved in the differentiated phenotype of smooth muscle cells. AB - The calcineurin-mediated pathway is involved in skeletal and cardiac hypertrophy and vascular development in vivo, but the relationship between this pathway and the phenotype of smooth muscle cells (SMCs) remains unknown. Using visceral SMCs in culture as a model system of differentiated SMCs, we investigated the role of the calcineurin-mediated pathway in maintaining the differentiated phenotype of SMCs, which depends on the insulin-like growth factor (IGF-I)-triggered activation of the phosphatidylinositol 3-kinase (PI3-K)/protein kinase B (PKB(Akt)) pathway. Treatment with calcineurin inhibitors, cyclosporin A or FK506, or the forced expression of the natural calcineurin inhibitor, CAIN, induced SMC dedifferentiation. Notably, suppression of the promoter activities of the SMC molecular markers caldesmon and alpha1 integrin by blocking the PI3 K/PKB(Akt) pathway was rescued by the forced expression of constitutively active calcineurin Aalpha, suggesting that the calcineurin-mediated pathway is critical for maintaining the differentiated phenotype of SMCs and works downstream of the PI3-K/PKB(Akt) pathway. PMID- 12535644 TI - Activated beta-catenin induces osteoblast differentiation of C3H10T1/2 cells and participates in BMP2 mediated signal transduction. AB - Wnt glycoproteins are important regulators of cellular differentiation and embryonic development. Some Wnt proteins induce stabilization of beta-catenin which cooperatively regulates gene expression with LEF/Tcf transcription factors. Here we demonstrate a direct role for beta-catenin signaling in osteoblast differentiation and in BMP2-mediated signal transduction. Similar to treatment with BMP-2 protein, ectopic expression of stabilized beta-catenin in C3H10T1/2 cells or activation of endogenous beta-catenin signaling with LiCl induces expression of alkaline phosphatase mRNA and protein, a defined marker of early osteoblast differentiation. Unlike BMP2 protein, stabilized beta-catenin does not induce osteocalcin gene expression, a marker of late osteoblast differentiation. BMP2-induced differentiation also leads to activation of endogenous beta-catenin signaling thus implicating beta-catenin in early steps of BMP2-mediated osteoblast differentiation. Effects of beta-catenin and BMP2 on C3H10T1/2 differentiation are not completely overlapping, implying that some aspects of BMP2-induced differentiation may be mediated by beta-catenin signaling and that beta-catenin can also participate in non-BMP2-dependent differentiation processes. PMID- 12535645 TI - Structural determinants of Rab and Rab Escort Protein interaction: Rab family motifs define a conserved binding surface. AB - Rab proteins are a large family of monomeric GTPases with 60 members identified in the human genome. Rab GTPases require an isoprenyl modification to their C terminus for membrane association and function in the regulation of vesicular trafficking pathways. This reaction is catalysed by Rab geranylgeranyl transferase, which recognises as protein substrate any given Rab in a 1:1 complex with Rab Escort Protein (REP). REP is therefore able to bind many distinct Rab proteins but the molecular basis for this activity is still unclear. We recently identified conserved motifs in Rabs termed RabF motifs, which we proposed to mediate a conserved mode of interaction between Rabs and REPs. Here, we tested this hypothesis. We first used REP1 as a bait in the yeast two-hybrid system and isolated strictly full-length Rabs, suggesting that REP recognises multiple regions within and properly folded Rabs. We introduced point mutations in Rab3a as a model Rab and assessed the ability of the mutants to interact with REP using the yeast two-hybrid system and an in vitro prenylation assay. We identified several residues that affect REP:Rab binding in the RabF1, RabF3, and RabF4 regions (which include parts of the switch I and II regions), but not other RabF regions. These results support the hypothesis that Rabs bind REP via conserved RabF motifs and provide a molecular explanation for the preferential recognition of the GDP-bound conformation of Rab by REP. PMID- 12535646 TI - A third human carnitine/organic cation transporter (OCTN3) as a candidate for the 5q31 Crohn's disease locus (IBD5). AB - Organic cation transporters function primarily in the elimination of cationic drugs in kidney, intestine, and liver. The murine organic cation/carnitine (Octn) transporter family, Octn1, Octn2, and Octn3 is clustered on mouse chromosome 11 (NCBI Accession No. NW_000039). The human OCTN1 and OCTN2 orthologs map to the syntenic IBD5 locus at 5q31, which has been shown to confer susceptibility to Crohn's disease. We show that the human OCTN3 protein, whose corresponding gene is not yet cloned or annotated in the human reference DNA sequence, does indeed exist and is uniquely involved in carnitine-dependent transport in peroxisomes. Its functional properties and inferred chromosomal location implicate it for involvement in Crohn's disease. PMID- 12535647 TI - Agouti-related protein has an inhibitory paracrine role in the rat adrenal gland. AB - alpha-Melanocyte-stimulating-hormone (alpha-MSH) is an agonist at the melanocortin 3 receptor (MC3-R) and melanocortin 4 receptor (MC4-R). alpha-MSH stimulates corticosterone release from rat adrenal glomerulosa cells in vitro. Agouti-related protein (AgRP) an endogenous antagonist at the MC3-R and MC4-R, is expressed in the adrenal gland. We investigated the expression of the MC3-R and MC4-R and the role of AgRP in the adrenal gland. MC3-R and MC4-R expression was detected in rat adrenal gland using RT-PCR. The effect of AgRP on alpha-MSH induced corticosterone release was investigated using dispersed rat adrenal glomerulosa cells. AgRP administered alone did not affect corticosterone release, but co-administration of AgRP and alpha-MSH attenuated alpha-MSH-induced corticosterone release. To investigate glucocorticoid feedback, adrenal AgRP expression was compared in rats treated with dexamethasone to controls. AgRP mRNA was increased in rats treated with dexamethasone treatment compared to controls. Our findings demonstrate that adrenal AgRP mRNA is regulated by glucocorticoids. AgRP acting via the MC3-R or MC4-R may have an inhibitory paracrine role, blocking alpha-MSH-induced corticosterone secretion. PMID- 12535648 TI - A kinase-inactive type II TGFbeta receptor impairs BMP signaling in human breast cancer cells. AB - Dominant negative receptor mutants are often utilized in order to abrogate signaling induced by growth factors. We have previously shown that expression of a dominant negative type II TGFbeta receptor (dnTbetaRII) in MDA-MB-231 breast cancer cells effectively abrogates TGFbeta signaling. In this letter, we report that expression of dnTbetaRII also impairs BMP2-mediated Smad1 phosphorylation as well as BMP2-mediated Smad-dependent transcriptional responses, resulting in an attenuation of BMP-mediated anti-proliferative effects. The fact that dnTbetaRII not only abrogates TGFbeta signaling but BMP signaling as well has important implications for the interpretation of data in which dominant negative mutants are utilized. PMID- 12535649 TI - A membrane potential-sensitive dye for vascular smooth muscle cells assays. AB - Changes in membrane potential of rat aorta smooth muscle cells were investigated using the bis-oxonol sensitive probe DIBAC2(3). We compared the changes in membrane potential induced by a high external KCl concentration in aorta smooth muscle cells from normotensive 2 kidney (2K) and from renal hypertensive 2 kidney 1 clip (2K-1C) rats. The spectral properties of the membrane potential were first characterized in aqueous buffers and in cultured smooth muscle cells from 2K and 2K-1C rat aortas. Fluorescence emission and the images were recorded using a laser scanning confocal microscope. The relationship between fluorescence intensity (FI) and membrane potential (psi(m)) as a function of the increasing extracellular KCl concentration was linear in the 5-40 mmol/L KCl range in both 2K and 2K-1C rat aorta cells. Cell membranes from 2K-1C rat aorta cells were more depolarized (-55 mV) than 2K rat aorta cells (-65 mV). The results show that in 2K-1C aorta cells only 10 mmol/L KCl was needed to induce complete membrane depolarization while in 2K cells 40 mmol/L KCl was needed to induce a similar effect. This study clearly shows that the method is suitable to measure the membrane potential in cultured smooth muscle cells. PMID- 12535650 TI - Dimerization of presenilin-1 in vivo: suggestion of novel regulatory mechanisms leading to higher order complexes. AB - A growing body of evidence indicates that presenilins could exist and be active as oligomeric complexes. Using yeast two-hybrid and cell culture analysis, we provide evidence that presenilin-1 (PS1) may self-oligomerize giving rise to specific full-length/full-length homodimers. When expressed in N2A and HEK239T cultured cells, full-length PS1-wt and 5(')myc-PS1-wt form specific homodimers corresponding to twice their molecular weight. The Alzheimer's disease-associated PS1 mutations Y115H, M146L, L392V, deltaE10(PS1(1-289/320-467)), the gamma secretase dominant negative mutant D257A, and the PS1 polymorphism mutant E318G do not affect their ability to self-oligomerize. Under non-denaturing conditions, endogenous PS1 forms specific homo-oligomers in human cultured cells. The results obtained herein suggest that PS1 associates intramolecularly to form higher order complexes, which may be needed for endoproteolytic cleavage and/or gamma secretase-associated activity. PMID- 12535651 TI - The role of maternal and zygotic Gprk2 expression in Drosophila development. AB - G protein-coupled receptor activity is controlled by a number of factors including phosphorylation by the family of G protein-coupled receptor kinases. This phosphorylation is an important first step in desensitization of the receptor. The role of G protein-coupled receptor kinases in cellular physiology has been extensively studied, but less is known about their role in development. A Drosophila G protein-coupled receptor kinase mutant (gprk2(6936)) has developmental defects throughout the life cycle of the fly. This allows the opportunity to address G protein-coupled receptor kinase's function in vivo. Using a series of transgenic flies in which the wild type Gprk2 gene is expressed under the control of the hsp70 or germline-specific promoter, in combination with germline mosaic analysis, we have made a detailed analysis of the developmental stages in which Gprk2 expression is required and the tissues that must express Gprk2 for rescue of the gprk2(6936) mutant. These studies have shown that Gprk2 expression is required in the germline for proper formation of the anterior egg structures and for early embryogenesis. In the absence of maternal Gprk2 activity, zygotic expression affords partial rescue of egg hatching, suggesting that Gprk2 also plays an important role in late embryogenesis. PMID- 12535652 TI - Nascent structure in the kinase anchoring domain of microtubule-associated protein 2. AB - Biological processes are often viewed as highly ordered interactions between well folded protein domains. The specific interactions exhibited by certain highly abundant neuronal proteins such as microtubule-associated protein 2 (MAP2) and tau stand in stark contrast because these proteins do not show evidence of structure by standard biophysical assays, yet they do bind to specific targets. It is conceivable that there are regions of MAP2 and tau with propensity to form structural domains upon binding a target. To search for evidence of such regions, limited proteolysis experiments were carried out on MAP2c, the smallest MAP2 isoform. Increased protease resistance was observed around the binding site for the RII subunit of cAMP-dependent protein kinase. Protein constructs spanning this region were produced based on the long-lived tryptic fragments Ser44-Arg93 and Ile94-Arg182, and were probed for structure using spectroscopic methods. The results support the existence of regions of nascent structure in the N-terminal region of MAP2c, which are believed to contribute to its regulatory function. PMID- 12535653 TI - Aspirin inhibits human coronary artery endothelial cell proliferation by upregulation of p53. AB - Aspirin (acetylsalicylic acid, ASA) is effective in the primary and secondary prevention of vascular events. This effect is mediated in large part by platelet inhibition; however, non-platelet-mediated effects may also be relevant in the overall efficacy of ASA. We determined the effect of ASA on the synthesis of DNA and total proteins in cultured human coronary endothelial cells (HCAECs). Fourth generation HCAECs were cultured and treated with ASA and rate of synthesis of DNA and total proteins was determined by incorporation of [3H]thymidine and [3H]proline, respectively. ASA inhibited DNA synthesis by 50% at a concentration of 1mM and protein synthesis by 50% at a concentration of 2mM. The inhibitory effect of ASA was observed as early as 2h after treatment of HCAECs. The inhibition of DNA and protein synthesis could be reversed within 24h after removal of the drug from the culture medium. Indomethacin also inhibited DNA and protein synthesis. Western blot analysis revealed that the expression of p53 protein was increased after treatment of the cells with ASA. These observations indicate that ASA decreases endothelial cell proliferation through cell cycle arrest mediated by enhanced p53 expression. Arrest of endothelial proliferation and activation may be an important mechanism of the beneficial effect of ASA in acute coronary syndromes. PMID- 12535654 TI - Identification of apoptosis-inducing factor in human coronary artery endothelial cells. AB - Apoptosis has been observed in vascular tissues in response to pro-inflammatory and pro-atherosclerotic stimuli, such as oxidized low density lipoproteins (ox LDL), angiotensin II (Ang II), and tumor necrosis factor-alpha (TNF-alpha). Apoptosis is believed to be mediated via caspase-dependent pathway. Recently, a 57 kDa molecule, apoptosis-inducing factor (AIF), has been described as a basis for cell injury via a caspase-independent pathway. This study was designed to identify the presence of AIF and the regulation of its gene expression in human coronary artery endothelial cells (HCAECs). Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to determine AIF mRNA and protein expression. Cultured HCAECs were treated with ox-LDL (10-40 microg/ml), angiotensin II (10(-9)-10(-6)M), or TNF-alpha (0.1-10n g/ml). AIF was barely detectable in unstimulated HCAECs; however, treatment with ox-LDL, but not with Ang II or TNF-alpha, significantly increased the expression of AIF in a concentration- and time-dependent manner. DNA sequencing analysis substantiated the existence of AIF in the HCAECs. Treatment of cells with the caspase inhibitor with Z-VAD-fmk did not change ox-LDL-mediated AIF protein expression. Ox-LDL mediated upregulation of AIF expression was inhibited by actinomycin D, suggesting transcriptional regulation. Further, upon treatment of cells with ox LDL AIF translocated from mitochondria to the nucleus, as determined by immunocytochemistry. These data suggest that AIF is expressed in HCAECs and is upregulated by ox-LDL. PMID- 12535655 TI - An increased expression of nucleolin is associated with a physiological nucleolar segregation. AB - Nucleolar segregation is the most striking cellular phenotypic feature of cold acclimatized carp and depicts the cyclical reprogramming that the physiology of the fish undergoes between summer and winter, where a clear differential expression of some nucleolar related genes occurs. We characterized carp nucleolin, a nucleolar protein involved in multiple steps of ribosome biogenesis, and evaluated its expression upon fish acclimatization. We show that the carp cDNA deduced amino acid sequence exhibits the same tripartite structural organization found in other species. Nevertheless, we observed that nucleolin mRNA expression was strongly induced in the cold-adapted carp as was the nuclear protein content, assessed by immunocytochemistry in liver sections. The physiological up-regulation of nucleolin in the cold-acclimatized carp, where rRNA transcription and processing are depressed concomitantly with the nucleolus segregation, is consistent with the notion that nucleolin plays a fundamental role in repressing rRNA synthesis. PMID- 12535656 TI - Differential calcium response in HeLa and HeLa-Fas cells by cytotoxic T lymphocytes. AB - We constructed a CD95 overexpressing HeLa cell line which was extremely sensitive towards CD95 mediated apoptosis. In these CD95 overexpressing cells, CD95 blocks the nuclear calcium signal induced by perforin positive and CD95 ligand positive killer cells. This phenomenon is highly relevant in states of inflammatory syndromes such as systemic inflammatory response syndrome (SIRS) and sepsis which are associated with a high probability to reactivate latent viruses due to a functional deficiency of cytotoxic effectors. PMID- 12535657 TI - Generation and characterisation of functional recombinant antibody fragments against RNA replicase NIb from plum pox virus. AB - A monoclonal antibody (mAb 2A) able to react against the RNA replicase NIb from plum pox virus (PPV) was obtained and used for generating a specific scFv fragment. The VH and VL coding sequences were cloned and expressed as a fusion scFv protein to alkaline phosphatase. This fusion protein was able to recognise viral NIb in both Western and tissue-print ELISA blots. The affinity and specificity of scFv2A for NIb was similar to that of the parental mAb and the region YLEAFY from PPV-NIb was identified by PEPSCAN assay as the putative epitope. Isolated VH domains from scFv2A were also expressed as fusion to alkaline phosphatase. However, their ability to react against NIb was greatly altered. scFv2A fragments were transiently expressed in the cytosol of Nicotiana benthamiana and although they accumulated to low levels, inhibition-ELISA results indicated that they retained antigen-binding activity. PMID- 12535658 TI - Molecular cloning and characterization of a novel cystatin-like molecule, CLM, from human bone marrow stromal cells. AB - The cystatins are physiological cysteine proteinase inhibitors. Here we report the cloning of a novel human cystatin-like molecule (CLM) from human bone marrow stromal cell (BMSC) cDNA library. The putative CLM protein contained 159 residues with a 29-residue signal peptide. CLM protein was highly homologous to family 2 cystatins, especially mouse and human testatin. The CLM gene spanned two exons and was mapped on chromosome 20p11.2, among cystatin superfamily gene clusters. CLM mRNA was barely detected in most tumor cell lines except for breast adenocarcinoma MCF-7 cells and glioblastoma U251 cells, but after LPS or PMA stimulation, CLM expression was increased in myelogenous leukemia cell lines HL 60 and U-937. Northern blot analysis revealed CLM was ubiquitously expressed in normal tissues, which was clearly different from the testis-specific expression pattern of most family 2 cystatins. When overexpressed in 293 cells, GFP-fused CLM targeted extracellularly through secretory pathway by Golgi apparatus. The results indicated that the secreted CLM protein might play roles in hematopoietic differentiation or inflammation. PMID- 12535659 TI - Novel drug designing approach for dual inhibitors as anti-inflammatory agents: implication of pyridine template. AB - Compounds incorporating thiophene moiety, a pi excess five membered heterocycle, have attracted a great deal of research interest, owing to the therapeutic utility of the template as useful drug molecular scaffolding. We report the synthesis and pharmacological evaluation of thiophenes substituted with 4 methanesulfonyl benzoyl moiety at the fifth position of the ring, as possible anti-inflammatory lead candidates. The aryl sulfonyl methyl thiophene analogs AP29, AP82, and AP37, when screened for anti-inflammatory activity in carrageenin induced rat paw edema, an acute in vivo model, exhibited moderate to good activity at a dose level of 100 mg/kg body weight P.o compared to Ibuprofen. In a five day formalin induced rat paw edema, a chronic in vivo anti-inflammatory model, candidates AP29, AP82, and AP37 inhibited the disease progression by 53%, 34%, and 65%, respectively on the fifth day, at a dose level of 100 mg/kg body weight P.o compared to Rofecoxib, Ibuprofen, and Dexamethasone at therapeutic doses which gave a protection of 53.8%, 81.5%, and 81.5%, respectively. The replacement of the 4-methanesulfonyl benzoyl moiety in AP82 with the pyridine template, 3,5-dimethyl-4-methoxy-2-pyridyl function, gave rise to AP84, which was less active in the acute model, but gave 54% and 75% protection both during the first day and fifth day, respectively, in the chronic model. A dual mechanism of action is proposed for AP84, a non-steroidal drug which has exhibited remarkable activity when compared to the steroid dexamethasone. These results open up new avenues in designing novel anti-inflammatory drugs as dual inhibitors with the incorporation of a pyridine template as part of the pharmacophore. PMID- 12535660 TI - Acetylation and level of mitochondrial transcription factor A in several organs of young and old rats. AB - To gain further information on the role of mitochondrial transcription factor A (TFAM) in mitochondrial biogenesis, we studied the post-translational modifications of the protein in 6- and 28-month-old rat liver. Mass spectrometry and immunoblot analysis revealed that TFAM was acetylated at a single lysine residue and that the level of acetylation did not change with age. The measurement of the content of TFAM and of mitochondrial DNA (mtDNA) in several organs (cerebellum, heart, kidney, and liver) of young and old rats showed an age related increase of mtDNA and TFAM in all the organs analyzed, except in heart. These data are discussed in the light of the multiple roles of TFAM in mitochondrial biogenesis and of the age-related change of the mitochondrial transcription. PMID- 12535661 TI - Inorganic tripolyphosphate (PPP(i)) as a phosphate donor for human deoxyribonucleoside kinases. AB - Inorganic tripolyphosphate (PPP(i)) and pyrophosphate (PP(i)) were examined as potential phosphate donors for human deoxynucleoside kinase (dCK), deoxyguanosine kinase (dGK), cytosolic thymidine kinase (TK1), mitochondrial TK2, and the deoxynucleoside kinase (dNK) from Drosophila melanogaster. PPP(i) proved to be a good phosphate donor for dGK, as well as for dCK with dCyd, but not dAdo, as acceptor substrate, illustrating also the dependence of donor properties on acceptor. Products of phosphorylation were shown to be 5(')-phosphates. In striking contrast to ATP, the phosphorylation reaction follows strict Michaelis Menten kinetics, with K(m) values of 74 and 92 microM for dCK and dGK, respectively, and V(max) values 40-50% that for ATP. With the other three enzymes, as well as for dCK with dAdo as acceptor, no, or only low levels (0.05). In addition, pre-ischemic bumetanide treatment reduced the ipsilateral water content increase by 70% (P<0.05). Inhibition of NKCC1 did not attenuate poly (ADP-ribose) polymerase cleavage or the number of TUNEL labeled apoptotic cells in ischemic brains. These results suggest that inhibition of NKCC1 attenuates cytotoxic edema and necrotic neuronal death during focal ischemia. Activation of NKCC1 activity plays a role in the early stage of ischemic damage. PMID- 12535775 TI - Opioid receptor activation attenuates nicotinic enhancement of spontaneous GABA release in lateral spiriform nucleus of the chick. AB - We examined the effects of opioids on the nicotinic enhancement of spontaneous GABA release from presynaptic terminals in the lateral spiriform nucleus (SpL) of the chick. Whole cell recordings from SpL neurons in brain slices were used to monitor spontaneous GABA release. Nicotine (1 microM) produced an 8-fold increase in the frequency of GABA events without changing their amplitude, consistent with an increase of GABA release from presynaptic terminals. L-enkephalin (1 microM) blocked these effects of nicotine on presynaptic GABA release, and the opioid antagonist naloxone (100 nM) antagonized the actions of L-enkephalin. The selective mu agonist DAMGO (300 nM) also attenuated the nicotine-mediated enhancement of GABA release, and the mu selective antagonist CTOP (1 microM) blocked the actions of DAMGO. In contrast, the kappa opioid agonist U50488 (3 microM) and the delta opioid agonist DPDPE (1 microM) had no effect. The results demonstrate that presynaptic release of GABA in the SpL can be regulated by both nicotinic agonists and mu opioids. While mu opioids have little effect on GABA release by themselves, they are able to block the marked enhancement of GABA release normally produced by nicotine. Since both cholinergic and enkephalinergic nerves are present in the SpL, the interactions of these two neurotransmitter systems may serve to precisely regulate GABA release in this brain region. PMID- 12535776 TI - Reduction in parasympathetic reflex vasodilatation following stereotaxic ear-bar insertion: importance of reduced afferent input. AB - As in our previous report, when cats were fitted with stereotaxic ear-bars 'type A' animals (26 out of 41) still exhibited a parasympathetic reflex lip blood flow (LBF) increase in response to lingual nerve stimulation, while in 'type B' animals (the remaining 15) it was greatly reduced or abolished. We compared (in both magnitude and in their sensitivity to hexamethonium, 10 mg/kg, i.v.) the LBF responses evoked by electrical stimulation of various sites within the reflex arc (lingual nerve, trigeminal ganglion, spinal trigeminal nucleus (Vsp)) in type A and type B animals to examine where the suppressive effect of ear-bar insertion might be exerted (using artificially ventilated, cervically vago-sympathectomized cats deeply anesthetized with alpha-chloralose and urethane). After ear-bar insertion: (a) in type A animals, stimulation of both lingual nerve and Vsp evoked a similar, hexamethonium-sensitive LBF increase; (b) in type B animals (in which lingual-nerve stimulation evoked no LBF increase), Vsp stimulation evoked a hexamethonium-sensitive LBF increase; (c) in both type A and type B animals, trigeminal ganglion stimulation consistently elicited an LBF increase (abolished by hexamethonium in type A, but reduced by only 50% in type B). These results suggest (i) that abolition of the lingual nerve-induced parasympathetic reflex vasodilatation by ear-bar insertion is due to reduced afferent traffic (in peripheral trigeminal or facial nerves) rather than to a damaged efferent output, and (ii) this effect in type B animals seems somehow to allow an antidromic (hexamethonium-insensitive) vasodilatation to occur on trigeminal ganglion stimulation. PMID- 12535777 TI - Presynaptic dopaminergic function is largely unaltered in mesolimbic and mesostriatal terminals of adult rats that were prenatally exposed to cocaine. AB - Fast-scan cyclic voltammetry in brain slices and postmortem tissue content assessment were used to evaluate presynaptic dopaminergic function in the caudate putamen and nucleus accumbens of adult male rats (180+ days old) that were prenatally treated with either cocaine or saline. Experiments were carried out to test whether there were differences in dopamine release, reuptake, autoreceptor function or the tissue levels of dopamine and its metabolites between cocaine- and saline-exposed rats. We report that presynaptic dopaminergic function remains largely intact in adult rats that were prenatally exposed to cocaine. The ability of terminals in the caudate putamen and nucleus accumbens to release and regulate dopamine is unaltered by prenatal cocaine exposure. However the tissue content of dopamine in the caudate putamen was decreased, representing a diminution in the dopamine storage pool. We conclude, therefore, that behavioral changes that have previously been observed in rats that were prenatally exposed to cocaine are not mediated through alteration of presynaptic dopaminergic mechanisms in these brain regions. PMID- 12535778 TI - Modulation of afferent inflow during the control of balancing tasks using the lower limbs. AB - This study investigated the control of sensory inputs during the performance of an inverted-pendulum balancing task. Experiments were conducted to examine modulation of proprioceptive inputs during balance tasks of varying difficulty. It was hypothesized that proprioceptive inputs to both spinal and cortical levels would be facilitated during a challenged balance task. In contrast, during challenged balance control, results revealed task-specific facilitation of sensory inputs to the cortex and inhibition of the spinal reflex pathway. Observations of increased transmission of proprioceptive inputs to the cortex and decreased transmission at the spinal level suggest that the cortex plays an important role in challenged balance, whereas the role for the spinal stretch reflex appears to be less important. PMID- 12535779 TI - A postnatal sensitive period for plasticity of cortical afferents but not cortical association fibers in rat piriform cortex. AB - Male and female rats underwent unilateral naris occlusion or sham surgery on either post-natal day (PN) 1 or after PN30. Following at least 30 days of unilateral olfactory deprivation, rats were urethane anesthetized and recordings were made from anterior piriform cortex (aPCX). Shock stimulation of afferent fibers (lateral olfactory tract) and association/commissural fibers evoked field potentials in aPCX that were analyzed across groups and between ages. The results demonstrate that early-onset unilateral olfactory deprivation depresses field potentials evoked by stimulation of the deprived cortical afferent, while late onset deprivation did not. In contrast, intracortical association fiber mediated field potentials in the deprived cortex were enhanced after both early-onset and late-onset deprivation. These results suggest differential developmental plasticity of afferent and association fiber pathways in paleocortex that mirrors that previously described in neocortical sensory systems. PMID- 12535781 TI - Immunohistochemical localization of the sigma1-receptor in oligodendrocytes in the rat central nervous system. AB - By using a new polyclonal antibody raised against a 21-amino acid peptide sequence corresponding to the fragment 138-157 of the cloned rat sigma(1) receptor, we demonstrated by immunoperoxidase and double immunofluorescence techniques, that rat oligodendrocytes express the sigma(1)-receptor. Experiments in vivo and in vitro showed that sigma(1)-receptor colocalized with specific markers of progenitor (A2B5) and mature oligodendrocytes (GalC, RIP). These results suggest that sigma(1)-receptor in oligodendrocytes might be involved in myelination by direct implication in cholesterol biosynthesis or by interaction with endogenous ligands such as neurosteroids. PMID- 12535780 TI - The association of beta-site APP cleaving enzyme (BACE) C786G polymorphism with Alzheimer's disease. AB - The deposition of amyloid beta-peptide (Abeta) plays a crucial role in the pathogenesis of Alzheimer's disease (AD). Beta-site APP cleaving enzyme (BACE) is the rate-limiting enzyme in the Abeta formation. BACE mutations/polymorphisms may be associated with AD. We searched the BACE coding region mutations/polymorphisms of cDNA in 25 AD patients and 100 healthy controls by single-strand conformational polymorphism. A polymorphism at BACE coding region was identified and confirmed to be 786C/G polymorphism by nucleotide sequencing. Based on these findings, we investigated the association of this polymorphism with the occurrence of AD by PCR-RFLP. A total of 98 AD patients along with 138 controls were recruited in the present study. The allele frequencies of the 786C/G polymorphism were 0.622 for C and 0.378 for G in AD. In controls, the C and G allele frequencies were 0.691 and 0.309, respectively. No significant association of this polymorphism with the occurrence of AD can be established. Larger sample size may be necessary to identify other potential mutations/polymorphisms among BACE gene. PMID- 12535782 TI - Decreased NPY innervation of the hypothalamic nuclei in rats with cancer anorexia. AB - Whether the decrease in food intake that occurs at the onset of anorexia in tumor bearing (TB) rats is related to a change in the hypothalamic neuropeptide Y (NPY) system was tested by comparing NPY expression in sham operated Fischer Control and anorectic TB rats. Coronal cryocut sections of their fixed brain were processed by the peroxidase-antiperoxidase method with NPY polyclonal antibodies. NPY-immunoreactive fibers were widely distributed throughout the forebrain, but were most prominent in the hypothalamic paraventricular, suprachiasmatic, arcuate and periventricular nuclei. NPY-immunoreactive neurons were visualized in Control and anorectic TB rats in the preoptic region, the arcuate nucleus, and occasionally in the lateral hypothalamus. Semiquantitative image analysis showed a significant decrease in the NPY immunostaining in some hypothalamic nuclei of the anorectic TB rats, most prominently in the supraoptic nucleus, the parvocellular portion of the paraventricular nucleus, and, to a lesser extent, the suprachiasmatic and arcuate nuclei. No changes in NPY innervation were seen in the ventromedial nucleus and the lateral hypothalamus. The data support the hypothesis of an altered hypothalamic NPY system at the onset of anorexia in TB rats and also reveal the hypothalamic nuclei through which NPY influences food intake. PMID- 12535783 TI - Influence of inhibitory and excitatory inputs on serotonin efflux differs in the dorsal and median raphe nuclei. AB - The dorsal (DRN) and median raphe nuclei (MRN) are two major sources of serotonergic projections to forebrain that are involved in regulation of behavioral state and motor activity, and implicated in affective disorders such as depression and schizophrenia. To investigate afferent influences on serotonergic neurons, this study compared the role of endogenous GABA and glutamate in the DRN and MRN using microdialysis and measurement of locomotor activity in freely behaving rats. Local infusion of the GABA(A) receptor antagonist bicuculline increased serotonin (5-HT) efflux in the DRN but not the MRN. In contrast, infusion of glutamate receptor antagonists produced larger decreases in 5-HT efflux in the MRN compared with the DRN. Moreover, glutamate receptor antagonists attenuated the increase in 5-HT efflux produced by GABA receptor blockade in the DRN. Thus, the disinhibitory effect of GABA blockers could be ascribed in part to an enhanced influence of glutamate. Measurements of locomotor activity indicate that changes in 5-HT were not simply correlated with behavioral activity induced by drug infusion. In summary, the role of inhibitory and excitatory afferents was strikingly different in the DRN and MRN. GABA afferents were the predominant tonic influence on serotonergic neurons in the DRN. In contrast, glutamatergic but not GABAergic afferents had a strong tonic influence on serotonergic neurons in the MRN. PMID- 12535784 TI - Cyclooxygenase-2 mRNA expression is associated with c-fos mRNA expression and transient water ADC reduction detected with diffusion MRI during acute focal ischemia in rats. AB - Cyclooxygenase-2 (COX-2) plays an important role in the development of injury during cerebral ischemia and inhibition of its activity can reduce infarct size. COX-2 expression during acute ischemia is caused by activation of post-synaptic glutamate receptors, which occurs during spreading depression and ischemic depolarization. Both of these phenomena cause a reduction in the apparent diffusion coefficient of water (ADC), which can be detected with diffusion weighted magnetic resonance imaging. The reduction is believed to be caused by cellular swelling that occurs as cells depolarize. The goal of this work was to determine the spatial relationship between cyclooxygenase-2 mRNA (cox-2) expression, c-fos mRNA expression and ADC reduction during acute focal cerebral ischemia. Adult rats were subjected to either 30- or 60-min permanent occlusion of the middle cerebral artery. A 2-Tesla scanner was used to acquire diffusion weighted echo-planar images throughout the ischemic period, which were used to calculate ADC maps. Cox-2 and c-fos mRNA were detected with (35)S in situ hybridization. The results indicate that, for rats subjected to 60-min ischemia, cox-2 was observed in superficial layers of cortex, where transient ADC reduction and c-fos expression were observed. The same was true for most rats subjected to 30-min ischemia. However, in a small number of rats of the 30-min group, cox-2 mRNA expression was observed in regions exhibiting transient and persistent ADC reduction with no c-fos expression. The results suggest that cox-2 mRNA expression during acute MCA occlusion is caused by either or both spreading depression and transient ischemic depolarization. PMID- 12535785 TI - Subtoxic concentration of manganese synergistically potentiates 1-methyl-4 phenylpyridinium-induced neurotoxicity in PC12 cells. AB - Endogenous or exogenous substances that are toxic to dopaminergic cells have been proposed as possible cause of idiopathic Parkinson's disease (PD). 1-Methyl-4 phenylpyridinium (MPP(+)) and manganese are dopaminergic neurotoxins causing a parkinsonism-like syndrome. Here, we studied the possible synergistic reaction between these two neurotoxins using rat PC12 pheochromocytoma cells. MPP(+) induced a delayed neurotoxicity in PC12 cells. Although low concentration of manganese did not cause cell damage, it markedly enhanced MPP(+)-induced neurotoxicity with characteristics of apoptosis, such as DNA laddering and activation of caspase-3. To understand the mechanism of enhancement of subtoxic concentration of manganese on MPP(+)-induced neurotoxicity, we investigated the reactive oxygen species (ROS) generation using a molecular probe, 2',7' dichlorofluorescein diacetate. Although subtoxic concentration of manganese alone did not induce ROS increase, it significantly enhanced the ROS generation induced by MPP(+). We also determined the intracellular MPP(+) content. A time- and concentration-dependent increase of MPP(+) levels was found in PC12 cells treated with MPP(+). The accumulation of MPP(+) by PC12 cells was not affected by manganese. Taken together, these studies suggest that co-treatment with MPP(+) and manganese may induce synergistic neurotoxicity in PC12 cells and that subtoxic concentration of manganese may potentiate the effect of MPP(+) by an ROS dependent pathway. PMID- 12535786 TI - A novel trivalent cation chelator Feralex dissociates binding of aluminum and iron associated with hyperphosphorylated tau of Alzheimer's disease. AB - Aluminum (Al(III)) and iron (Fe(III)) are reported to accumulate in neurofibrillary tangles of the Alzheimer's disease (AD) brain. In these lesions Al (III) and Fe (III) bind with hyperphosphorylated tau (PHFtau), the major constituent of the lesions, and induce its aggregation. It is thought that inhibition and dissociation of such Al (III)/Fe (III) binding associated with PHFtau could slow or halt the tau-related neurofibrillary degeneration in patients with AD. A study, using a previously developed in vitro system in which Al (III) and Fe (III) interact with PHFtau on AD brain sections and on immunoblot membranes showed that the potent Al (III)/Fe (III) chelator desferrioxamine elicited Al (III) chelation when subjected to autoclave heating. Here, the ability of a recently developed chemical chelator Feralex-G to remove PHFtau bound Al (III)/Fe (III), using reaction conditions at 37 degrees C, was examined and compared with that of desferrioxamine. Chelation of Fe(III) was achieved by both compounds with no discernible difference in their chelating ability. In contrast, in the present system, the two chelators gave a different Al (III) chelation response. When incubated at 37 degrees C, desferrioxamine failed to attain notable Al (III) chelation, while Feralex-G displayed efficient Al (III) chelation. Thus, when considering competitive Al (III) removal from brain PHFtau, Feralex-G is a stronger chelator for Al(III) than desferrioxamine. The efficient Al (III) chelation attainable by Feralex-G adds weight to its potential clinical usefulness as a medicine in the aluminum/iron chelation therapy for patients with AD. PMID- 12535787 TI - Formalin- or adjuvant-induced peripheral inflammation increases neurokinin-1 receptor gene expression in the mouse. AB - Substance P (SP) has been widely studied as a mediator of nociception. The release of SP from primary afferent neurons is increased during nociception, and SP activates neurokinin-1 (NK-1) receptors in the spinal cord and periphery. Nociception-evoked alterations in NK-1 receptor gene expression have been studied in rat models of persistent pain but have not been characterized in any murine models of peripheral inflammation. This study assessed behavioral responses and NK-1 receptor mRNA gene expression in mice receiving formalin or Freund's complete adjuvant (CFA) as an inflammatory stimulus. Mechanical withdrawal thresholds were measured before injection of formalin or CFA and hind paw licking/biting timed during the late-phase of the formalin response. Two and 24 hours after formalin or CFA injection, mechanical withdrawal thresholds were measured and the mice euthanized. Solution hybridization-nuclease protection assays were used to quantify NK-1 receptor mRNA levels. Results demonstrated that inflamed hind paws were edematous, and the withdrawal thresholds of the inflamed hind paws were significantly lower after formalin or CFA injection. Neurokinin-1 receptor mRNA levels in the ipsilateral dorsal spinal cords of mice were higher at 24 h after formalin injection or 4 days after CFA injection. These results confirm that mice are hyperalgesic at late time points after formalin or adjuvant injection when NK-1 receptor gene expression is elevated in the dorsal spinal cord. This supports the hypothesis that increased NK-1 receptor gene expression contributes to the development and maintenance of a hyperalgesic state. PMID- 12535788 TI - Orientation mosaic in barn owl's visual Wulst revealed by optical imaging: comparison with cat and monkey striate and extra-striate areas. AB - Using the technique of intrinsic signal optical imaging, orientation preference maps were obtained from the Wulst of the barn owl in the area that represents central vision, and from the visual cortices (V1 and V2) of cat and marmoset monkey. Iso-orientation domains in barn owl's visual Wulst were patch-like structures with an inter-patch distance of approximately 0.9 mm, arranged in a pinwheel-like manner around singularity points. The size of the iso-orientation domains in barn owl was larger than those found in area V1, but comparable to those found in area V2, of cat and monkey. Superficial layers of the owl visual Wulst may be equivalent to extra-striate visual areas of primates and carnivores, as already suggested by electrophysiologists discussing the much increased radial dimensions of the Wulst compared with neocortex in mammals. PMID- 12535789 TI - Maternal stress differently alters nociceptive behaviors in the formalin test in adult female and male rats. AB - The long-term effects of restraint stress in Wistar rats during the last week of gestation were investigated on the acute and tonic phases of the specific biphasic nociceptive behavioral response in the formalin test in offspring, females and males, at 90 days. Prenatal stress produced significant changes in formalin-induced pain, which was more pronounced in females as compared to males. The distorted response in females was more at the supraspinal level with an increased intensity of the licking response in both phases as well as with an increased their duration. Results concerning changes of the interphase length indicate the impairments of inhibitory mechanisms in the central nervous system. Furthermore, profound difference in the effects of prenatal stress on the first phase but similarity in these effects on the second phase in females and males are indirect strong support of the view that the second phase in the formalin test can not be mediated by central sensitization alone but greatly depends on signals ongoing from nociceptive primary afferents. Finally, the results obtained in males are important argument in favor of assumption about different mechanisms of acute and tonic pain. Taken together, these studies show that prenatal stress alters nociceptive behaviors in the formalin test in rats at 90 days in a sex specific manner. PMID- 12535790 TI - Fox odour affects corticosterone release but not hippocampal serotonin reuptake and open field behaviour in rats. AB - Group-housed Sprague-Dawley (SD) rats exposed for 1 h to 2,5-dihydro-2,4,5 trimethylthiazoline (TMT, a component of fox feces) did not display changes in hippocampal serotonin (5-HT) metabolism and [3H]5-HT reuptake, compared to water or butyric acid. Such an observation extended to isolated SD and Fischer 344 rats. When group-housed SD rats were tested 1 week after a 1-h exposure to TMT, hippocampal 5-HT metabolism, [3H]5-HT reuptake, and [3H]paroxetine binding at the 5-HT transporter remained unchanged. This study questions TMT as a specific predatory stimulus as both butyric acid and TMT increased plasma corticosterone levels whilst leaving intact open field behaviour (at least in group-housed SD rats). PMID- 12535791 TI - Voltage-dependent Ca2+ currents in rat cardiac dorsal root ganglion neurons. AB - This study presents the kinetic and pharmacological properties of voltage-gated Ca(2+) currents in anatomically defined cardiac dorsal root ganglion (DRG) neurons in rats. The neurons were labelled by prior injection of fluorescent tracer Fast Blue into the pericardial sack. There were three distinct groups of neurons with respect to cell size: small (27% of total; cell capacitance <30 pF), medium (65% of total; capacitance 30-80 pF) and large neurons (8% of total; capacitance >80 pF). The properties of Ca(2+) currents were tested in small and medium-sized neurons. In large neurons currents could not be adequately controlled and were not analysed. Ca(2+) currents did not completely inactivate during 100 ms depolarising voltage steps. The activation thresholds in small ( 36.9+/-1.3 mV) and medium (-39.0+/-1.3 mV) size neurons were similar. Current densities were 105.8 pA/pF in small and 97.4 pA/pF in large neurons and also did not differ. The kinetic properties of activation and inactivation did not differ between small and medium-sized cardiac DRG neurons. At membrane potentials between -50 and -60 mV (the expected resting membrane potential in these neurons) 55 to 70% of Ca(2+) currents in small and medium-sized neurons were available for activation. Both, small and medium-sized neurons expressed similar proportions of L (7.5%), N (25%) and P/Q (36%) type Ca(2+) currents. We conclude that small and medium-sized cardiac DRG neurons are homogeneous with respect to the expression and properties of voltage-gated Ca(2+) currents. Voltage-gated Ca(2+) currents probably play an important role in action potential generation in cardiac DRG neurons due to their availability for activation at resting membrane potential, their high density and voltage threshold close to the threshold for voltage-gated Na(+) currents. PMID- 12535793 TI - Chemokine receptors in inflammation: an overview. AB - Chemokine receptors play a key role in directing the migration of inflammatory cells into various injured or infected organs. However, migration of inflammatory cells into tissues can in itself be a cause and amplifier of tissue damage and disease, particularly in chronic autoimmune or allergic disorders. On this basis, much effort is currently devoted at the identification of molecular signals regulating the recruitment of inflammatory cells into tissues and at developing novel strategies to inhibit discrete pathways in this process. Great progress has recently been made in identification of a number of chemokine receptors involved in the process of leukocyte migration. The challenge is now to elucidate the specific contribution and involvement of the different receptors in distinct inflammatory processes and diseases and to prove that interference with any of these pathways may lead to development of novel therapeutics. PMID- 12535794 TI - The study of CXCR3 and CCR7 pharmacology using [35S]GTPgammaS exchange assays in cell membranes and permeabilized peripheral blood lymphocytes. AB - The GTPgammaS exchange assay is a functional model corresponding to the first step of G protein-coupled receptor activation. We provide simple methodologies and controls for setting up GTPgammaS exchange assays in both cell membranes and permeabilized peripheral blood lymphocytes. Specifically, we use guanosine 5' [35S]triphospate ([35S]GTPgammaS) exchange, in concert with radioligand binding assays, to assess the expression and function of two chemokine receptors important in the trafficking of T lymphocytes: CXCR3 and CCR7. The studies presented here illustrate the utility of GTPgammaS exchange assays in the study of chemokine receptor pharmacology. PMID- 12535795 TI - Development and characterisation of tetracycline-regulated phosphoinositide 3 kinase mutants: assessing the role of multiple phosphoinositide 3-kinases in chemokine signaling. AB - A combination of pharmacological, biochemical, molecular and genetic evidence supports a key role for phosphoinositide 3-kinase (PI3K) and its associated signalling cascade in cell migration in response to members of the chemokine family. PI3Ks can be divided into three main classes on the basis of their in vitro lipid substrate specificity, structure and likely mode of regulation. The prototypical class I PI3Ks are heterodimers consisting of the class I(A) 85-kDa regulatory/adaptor subunit and a catalytic 110-kDa subunit and the class I(B) PI3K (PI3Kgamma), which is stimulated by G protein betagamma subunits. Whilst genetic evidence supports a key role for PI3Kgamma in mediating chemotactic responses, it is clear that other PI3K isoforms can be activated by chemokines and can potentially contribute to the chemotactic responses to chemokines. In order to get a more accurate picture of the precise role of individual PI3Ks in functional responses to chemokines, we report development of tetracycline inducible dominant-negative constructs of the class I(A) and class I(B) PI3Ks and their expression in the leukemic T cell line Jurkat. SDF-1/CXCR4-mediated chemotaxis of Jurkat cells is strongly, but incompletely abrogated (e.g. approximately 60-70%) in clones expressing the dominant-negative PI3Kgamma construct. Interestingly, Jurkat cells expressing a dominant-negative mutant of class I(A) PI3K also exhibited marked abrogation of chemotactic responses to SDF 1, albeit to lesser extent (e.g. approximately 30-40% inhibition) than observed with the class I(B) mutant. These data suggests that both class I(A) and class I(B) isoforms can contribute to chemotactic responses, and both are required for optimal migratory responses to SDF-1. Furthermore, neither isoform alone is able to support optimal migration in the absence of the other. This may reflect an important interplay between the two different forms of PI3K that has yet to be fully elucidated. The use of inducible expression systems such as that described here will be an important approach in assessing the role of not only individual PI3Ks, but also their downstream effector proteins, in supporting actin polymerisation and cytoskeletal rearrangements as well as chemotaxis and adhesion molecule up-regulation. PMID- 12535796 TI - Detecting rapid and transient upregulation of leukocyte integrin affinity induced by chemokines and chemoattractants. AB - The majority of integrins expressed on circulating leukocytes are quiescent and do not mediate adhesion. In the process of emigrating from blood into tissues, leukocytes are exposed to a variety of stimuli, including chemokines, which rapidly activate integrin function through changes in affinity and/or avidity. High affinity is brought about by a change in integrin conformation and results in formation of a stable bond with ligand. This can be measured in soluble ligand binding assays. Integrin conformation changes also result in exposure of previously masked epitopes that can be detected with monoclonal antibodies. In this report, methods used to detect high affinity integrins are reviewed. Four different approaches are discussed, including detection of masked epitopes with monoclonal antibodies, and binding of soluble ligands, ligand-coated beads, and ligand-mimetic peptides. Improved techniques to measure rapid changes in integrin affinity in real time may be valuable both to study mechanisms of inside-out integrin activation and as a tool to screen for effective inhibitors of integrin activation in inflammatory disease models. PMID- 12535797 TI - A real time in vitro assay for studying leukocyte transendothelial migration under physiological flow conditions. AB - The mechanisms underlying leukocyte migration across endothelial barriers are largely elusive. Most of the current knowledge on transendothelial migration (TEM) of leukocytes has been derived from in vitro modified Boyden chamber transfilter migration assays. In these assays, leukocyte migration towards chemokine gradients constructed across the endothelial barrier is measured under shear-free conditions. These assays do not incorporate the contribution of shear flow to leukocyte adherence and migration across the endothelial barrier. Furthermore, transfilter assays do not reconstitute the physiological distribution of endothelial chemokines shown to be displayed in vivo at high levels on vessel walls. To overcome these two drawbacks, we have recently developed a novel in vitro assay to follow real time leukocyte migration across endothelial barriers under physiological flow conditions. Using this assay, we have found that apically displayed endothelial chemokines could trigger robust lymphocyte TEM through signaling to lymphocyte-expressed G-protein coupled receptors. This migration required continuous exposure of lymphocytes, adherent to the endothelial barrier, to fluid shear, but did not require a chemotactic gradient across the barrier. In the present review, we describe this new flow based migration assay and discuss future applications for investigating TEM processes of different types of leukocytes across distinct endothelial barriers. PMID- 12535798 TI - In situ binding assay for studying chemokine interactions with endothelial cells. AB - The association of chemokines with endothelial cells (EC) and extracellular matrices is required for the prototypical pro-emigratory and pro-migratory in vivo activity of these molecules, respectively. In order to investigate chemokine binding to intact microanatomical structures, e.g. venular EC, we have developed an in situ binding assay. This is an autoradiographic morphological method in which the saturable binding of radiolabeled chemokines is studied in vitro in pieces of viable tissues. This article discusses the general applicability, advantages and shortcomings of the in situ binding assay in comparison with the other techniques available for visualizing chemokine receptor binding by cells in the tissues. We used this assay to demonstrate: (a) selective specific binding of CXC and CC chemokines to the EC of postcapillary venules but not capillaries or arteries; (b) selective specific binding of CC chemokines to the EC of afferent lymphatic vessels; and (c) selective specific binding of inflammatory chemokines to the EC lining high endothelial venules (HEV) in lymph nodes. The assessment of ligand cross-competition provided a fingerprint of chemokine-binding specificity of the EC in each of these microanatomical sites. This fingerprint could be paralleled with the chemokine-binding profiles of two non-signaling chemokine binding molecules, Duffy antigen receptor for chemokines (DARC) and D6, present in venular and lymphatic EC, respectively. These observations allowed us to put forward the hypotheses regarding the involvement of EC DARC and D6 in chemokine transport and presentation by the EC. PMID- 12535799 TI - Knock out models to dissect chemokine receptor function in vivo. AB - Chemokines are a family of small proteins involved in numerous biological processes ranging from hematopoiesis, angiogenesis and lymphocyte trafficking to the extravasation and tissue infiltration of leukocytes in response to inflammatory agents, tissue damage and bacterial or viral infection. Chemokines exert their effects by binding to specific G-protein-coupled seven-transmembrane receptors. In vitro studies suggest that the chemokine system is highly redundant in that most chemokines bind to more than one receptor and most receptors bind multiple chemokines. Therefore, targeted deletion of chemokine receptors has proved to be a useful tool for determining the distinct biological role of these molecules in vivo. PMID- 12535800 TI - Analysis of the role of chemokines in angiogenesis. AB - Chemokines, a large family of inflammatory cytokines, have been shown to play a critical role in the regulation of angiogenesis during several pathophysiologic processes, such as tumor growth, wound healing and ischemia. Semiquantitative or quantitative angiogenesis assays are commonly utilized to screen the angiogenic or angiostatic activity of chemokines. These include in vitro endothelial cell activation assays and ex vivo or in vivo models of neovascularization. Chemokines may exert their regulatory activity on angiogenesis directly or as a consequence of leukocyte infiltration and/or the induction of growth factor expression. The effect of chemokines on endothelium can be assessed by performing in vitro assays on purified endothelial cell populations or by in vivo assays. Nevertheless, each model used to evaluate the angiogenic or angiostatic activity of a discrete factor has advantages and limitations. Thus, in order to avoid under- or overestimating the regulatory effect of chemokines on angiogenesis and to evaluate all aspects of the angiogenic process, multiple assays are usually performed. This review summarizes past and recent studies on chemokines as modulators of angiogenesis with particular emphasis on the methods currently used for the assessment of chemokine-mediated angiogenic or angiostatic responses. PMID- 12535801 TI - Analysis of homing receptor expression on infiltrating leukocytes in disease states. AB - Chemokines represent a large family of polypeptides that signal through G-protein coupled receptors and have a role in chemotaxis, leukocyte homing, inflammation, hematopoiesis, angiogenesis and tumor growth. The chemokine/chemokine receptor system acts in coordination with a complex cytokine network to elicit and direct leukocyte infiltration into the inflamed tissue. In addition to promoting movement into the inflamed tissue, the chemokine/chemokine receptor system may also activate infiltrating cells, such as neutrophils and eosinophils, and induce local damage. In recent years, the elucidation of intricate chemokine networks has led to the identification of potential target molecules for therapeutic intervention. Of considerable interest has been the role of chemokine/chemokine receptors in regulating allergic lung inflammation. In this review, techniques to study in situ expression of chemokine receptors in inflamed tissues are presented and discussed. PMID- 12535802 TI - New models of intravital microscopy for analysis of chemokine receptor-mediated leukocyte vascular recognition. AB - Chemokines control selective targeting of circulating leukocytes to the microvasculature by triggering inside-out signal transduction pathways leading to integrin-dependent adhesion. Presently, a few examples of presentation of chemokines by the inner surface of blood vessels responsible for triggering leukocyte arrest are available. A complete pattern of chemokine expression and presentation by the endothelium in different tissues and in homeostatic versus inflammatory conditions is still lacking. This review describes a novel intravital microscopy model allowing visualization of cerebral vessels through the skull, and analysis of the interactions between different leukocyte subpopulations and the endothelium in brain superficial microvasculature of mice. It has been recently shown that inflamed brain endothelium expresses a combination of adhesion ligands and activating factor(s) for G(i)-linked receptors that together mediate lymphocyte recruitment, and that the combination of molecules involved in this central nervous system (CNS) venule adhesion cascade strongly favors the arrest of activated versus nai;ve T cells. However, the identification of the chemokine(s) expressed by the endothelium leading to integrin activation and arrest still remains to be determined. The new approach presented here to study the brain microcirculation may provide an useful tool for further investigations of physiologic and pathologic events that occur in the CNS. PMID- 12535803 TI - The SCID-hu Skin mouse as a model to investigate selective chemokine mediated homing of human T-lymphocytes to the skin in vivo. AB - Here we report the establishment of an in vivo mouse model that allows monitoring of human T cell migration into human skin. This model is based on the use of severe combined immunodeficiency (SCID) mice transplanted with human skin (SCID hu Skin mice). Adoptively transferred human T helper (Th)2 cells obtained from atopic dermatitis skin lesions or peripheral blood T cells selectively migrate to the human skin grafts of these SCID mice in response to defined chemokines locally injected in the human skin grafts. Homing of human T cells into the human skin on SCID-hu Skin mice is a specific process since it only occurs in response to chemokine ligands that are specific for the chemokine receptors expressed on the migrating T cells. This mechanistic model allows analysis of the relevant steps involved in human T-lymphocyte migration into inflamed skin. In addition, it is successfully used for preclinical testing of drug candidates that are highly selective for human target molecules associated with the different steps of T cell migration in an environment that resembles the physiologic or pathologic conditions occurring in man. PMID- 12535804 TI - Myocardial laser revascularization for the treatment of end-stage coronary artery disease. AB - Myocardial laser revascularization is a novel therapeutic technique aimed at delivering oxygenated blood via a series of channels to the ischemic regions of the heart. These channels may be created surgically or via a less invasive percutaneous approach. In patients with end-stage coronary artery disease, both transmyocardial laser revascularization (TMR) and percutaneous myocardial laser revascularization (PMR) have been associated with a reduction in symptoms, improved exercise tolerance, and enhanced quality of life. However, the mechanism of action of laser therapy is incompletely understood, the results of objective cardiac perfusion measurements are inconclusive, and multiple randomized trials have failed to demonstrate an increase in survival. In addition, the positive results seen in TMR trials have been questioned because of a lack of blinding, raising the possibility that the benefit may have been due to the placebo effect. Finally, two recent sham-controlled, randomized clinical trials of PMR have not shown any benefit of the procedure, but instead have highlighted the important role of the placebo effect in the response to PMR. Further research is, therefore, needed to elucidate the value of myocardial laser revascularization. PMID- 12535805 TI - Sirolimus-eluting stent for treatment of complex in-stent restenosis: the first clinical experience. AB - OBJECTIVES: In this study, we assess the value of sirolimus eluting stent (SES) implantation in patients with complex in-stent restenosis (ISR). BACKGROUND: The treatment of ISR remains a therapeutic challenge, since many pharmacological and mechanical approaches have shown disappointing results. The SESs have been reported to be effective in de-novo coronary lesions. METHODS: Sixteen patients with severe, recurrent ISR in a native coronary artery (average lesion length 18.4 mm) and objective evidence of ischemia were included. They received one or more 18 mm Bx VELOCITY SESs (Cordis Waterloo, Belgium). Quantitative angiographic and three-dimensional intravascular ultrasound (IVUS) follow-up was performed at four months, and clinical follow-up at nine months. RESULTS: The SES implantation (n = 26) was successful in all 16 patients. Four patients had recurrent restenosis following brachytherapy, and three patients had totally occluded vessels preprocedure. At four months follow-up, one patient had died and three patients had angiographic evidence of restenosis (one in-stent and two in lesion). In-stent late lumen loss averaged 0.21 mm and the volume obstruction of the stent by IVUS was 1.1%. At nine months clinical follow-up, three patients had experienced four major adverse cardiac events (two deaths and one acute myocardial infarction necessitating repeat target vessel angioplasty). CONCLUSIONS: The SES implantation in patients with severe ISR lesions effectively prevents neointima formation and recurrent restenosis at four months angiographic follow-up. PMID- 12535806 TI - Variation in the alpha2B-adrenoceptor gene as a risk factor for prehospital fatal myocardial infarction and sudden cardiac death. AB - OBJECTIVES: Our aim was to corroborate the observed association between the deletion/deletion (DD) genotype of the insertion/deletion polymorphism in the alpha(2B)-adrenoceptor (AR) and increased risk for acute myocardial infarction (AMI), and to study whether this genotype also confers an increased risk for sudden cardiac death (SCD). BACKGROUND: Vasospasm has been suggested to play a role in AMI. Alpha(2)-AR mediate coronary vasoconstriction in humans, and studies on mice suggest the involvement of the alpha(2)-AR subtype B in vasoconstriction. A deletion variant of the human alpha(2B)-AR has been associated with impaired receptor desensitization in vitro. In a population-based prospective study of 912 middle-aged men, the DD genotype of the alpha(2B)-AR conferred an increased risk for AMI. METHODS: A series of 700 unselected sudden out-of-hospital deaths of middle-aged white men subjected to medico-legal autopsy was analyzed. RESULTS: Genotype information was obtained for 683 men (DD = 22%, insertion/deletion = 51%, insertion/insertion = 27%). Carriers of the DD genotype had an increased risk for SCD (n = 278, odds ratio [OR] = 2.0, p = 0.01) and fatal AMI (n = 84, OR = 2.1, p = 0.04) compared with the other two genotypes combined. The risks for SCD and fatal AMI were higher in carriers of the DD genotype who died before the age of 55 years (OR = 4.5 and 5.0, p < 0.001 for both). CONCLUSIONS: Middle-aged white men carrying the DD genotype of the alpha(2B)-AR have a significantly increased risk for SCD and AMI, especially before the age of 55 years. PMID- 12535807 TI - Emerging importance of alpha-adrenergic coronary vasoconstriction in acute coronary syndromes and its genetic background. PMID- 12535808 TI - Analysis and results of prolonged resuscitation in cardiac arrest patients rescued by extracorporeal membrane oxygenation. AB - OBJECTIVES: We conducted this study to determine the result of prolonged cardiopulmonary resuscitation (CPR) with extracorporeal membrane oxygenation (ECMO) and the predictive factors for hospital discharge and ECMO weaning. BACKGROUND: Prolonged CPR carries considerable associated mortality and morbidity. As yet, ECMO for prolonged CPR has no definite results. Only small groups of patients and no detailed analysis have been reported. METHODS: Candidates for ECMO resuscitation were patients in cardiac arrest receiving CPR >10 min without return of spontaneous circulation and no absolute contraindication. Venoarterial ECMO was set up during CPR. We reviewed the data of 57 prolonged CPR patients who received ECMO during CPR over a six-year period. RESULTS: The mean duration of CPR was 47.6 +/- 13.4 min and that of ECMO was 96.1 +/- 87.9 h. The rate of weaning was 66.7%, and the survival rate was 31.6%. Multiple-organ failure was the major reason for mortality, despite successful weaning. Among survivors, long-term follow-up revealed 88.9% survival, and only 5.6% had a severe neurologic deficit. The results indicate that a shorter CPR duration, postcardiotomy arrest, myocardial indicators, a hepatic indicator, and lactic acid are significantly correlated with both weaning and survival, whereas late damage (level on the third or seventh day of reperfusion) rather than initial damage (level on the first day) was more predictive of the results. CONCLUSIONS: Prolonged CPR rescue by ECMO provides an acceptable survival rate and outcome in survivors. Our results of the selected cases encourage further investigations of the wider application of ECMO in CPR. PMID- 12535809 TI - RITZ-5: randomized intravenous TeZosentan (an endothelin-A/B antagonist) for the treatment of pulmonary edema: a prospective, multicenter, double-blind, placebo controlled study. AB - OBJECTIVES: The objective of this study was to evaluate the addition of intravenous (IV) tezosentan to standard therapy for patients with pulmonary edema. BACKGROUND: Tezosentan is an IV nonselective endothelin (ET)-1 antagonist that yields favorable hemodynamic effects in patients with acute congestive heart failure (CHF). METHODS: Pulmonary edema was defined as acute CHF leading to respiratory failure, as evidenced by an oxygen saturation (SO(2)) <90% by pulse oxymeter despite oxygen treatment. All patients received oxygen 8 l/min through a face mask, 3 mg of IV morphine, 80 mg of furosemide, and 1 to 3 mg/h continuous drip isosorbide-dinitrate according to their blood pressure level and were randomized to receive a placebo or tezosentan (50 or 100 mg/h) for up to 24 h. RESULTS: Eighty-four patients were randomized. The primary end point, the change in SO(2) from baseline to 1 h, was 9.1 +/- 6.3% in the placebo arm versus 7.6 +/- 10% in the tezosentan group (p = NS). The incidence of death, recurrent pulmonary edema, mechanical ventilation, and myocardial infarction during the first 24 h of treatment was 19% in both groups. Reduced baseline SO(2), lower echocardiographic ejection fraction, high baseline mean arterial blood pressure (MAP), and inappropriate vasodilation (MAP reduction at 30 min of <5% or >30%) correlated with worse outcomes. A post-hoc analysis revealed that the outcome of patients who received only 50 mg/h tezosentan was better than patients in the placebo group whereas patients receiving 100 mg/h had the worst outcomes. CONCLUSIONS: In the present study, tezosentan (an ET-1 antagonist) did not affect the outcome of pulmonary edema, possibly because of the high dose used. PMID- 12535810 TI - Complications of inappropriate use of spironolactone in heart failure: when an old medicine spirals out of new guidelines. AB - OBJECTIVES: This study was designed to investigate the appropriateness and complications of the use of spironolactone for heart failure (HF) in clinical practice. BACKGROUND: Spironolactone was reported by one prospective randomized trial to decrease morbidity and mortality in patients with New York Heart Association (NYHA) class III and IV HF. With this report (Randomized Spironolactone Evaluation Study [RALES] trial), we noted a marked increase in widespread use of spironolactone in patients with HF. Long-term outcome data with respect to safety and utilization of this medication in HF are not available. METHODS: To investigate the use of spironolactone for HF in a clinical setting, we analyzed the application of the RALES trial protocol to the care of 104 patients, whom we identified as being started on spironolactone for HF after prerelease of the RALES trial. RESULTS: We found broader use, less intensive follow-up, and increased complications with spironolactone treatment compared with the RALES trial. Cardiologists provided more appropriate care than did primary care providers. CONCLUSIONS: These data suggest that spironolactone is being used widely in HF without consideration of the NYHA class and ejection fraction, and without optimization of background treatment with angiotensin converting enzyme inhibitors and beta-blockers. Clinical follow-up does not adhere to the RALES trial guidelines, resulting in higher complications. We conclude that long-term studies with further safety and efficacy data are needed. PMID- 12535811 TI - Spironolactone in chronic heart failure:all's well that ends well. PMID- 12535812 TI - Gender, age, and heart failure with preserved left ventricular systolic function. AB - OBJECTIVES: This study was designed to determine if women are more likely than men to have heart failure (HF) with preserved systolic function after adjustment for potential confounders, including age. BACKGROUND: Although prior evidence suggests an independent association between female gender and preserved left ventricular systolic function (LVSF) in patients with HF, existing studies are limited by referral biases, small sample sizes, or the inability to adjust for a wide range of potential confounding variables. METHODS: This is a cross-sectional study using data from retrospective medical chart abstraction of a national sample of Medicare beneficiaries hospitalized with the principal discharge diagnosis of HF in acute-care nongovernmental hospitals in the U.S. between April 1998 and March 1999. Patients were eligible for this analysis if they were age 65 years or older, had documentation of LVSF, and corroboration of the diagnosis of HF. We used multivariable logistic regression to identify the correlates of preserved LVSF, which was defined as qualitatively normal function or quantitatively reported ejection fraction > or =0.50. Stratified regressions by gender were performed to identify significant interactions. RESULTS: Of the 19,710 patients in the analysis, preserved LVSF was present in 6,700 (35%), 79% of whom were women. In contrast, among the 12,956 patients with impaired LVSF, only 49% were women. Patients with preserved LVSF were 1.5 years older than those with impaired LVSF. After adjustment for age and other patient factors, female gender remained strongly associated with preserved LVSF (calculated risk ratio = 1.71; 95% confidence interval 1.63 to 1.78). The association was consistent in all age groups, and was similar in patients with or without coronary artery disease, hypertension, pulmonary disease, renal insufficiency, or atrial fibrillation. CONCLUSIONS: In elderly patients hospitalized with HF, preserved systolic function is primarily a condition of women, independent of important demographic and clinical characteristics. PMID- 12535813 TI - The less familiar face of heart failure. PMID- 12535814 TI - Avoidance of the left lateral decubitus position during sleep in patients with heart failure: relationship to cardiac size and function. AB - OBJECTIVES: We sought to determine whether patients with congestive heart failure (CHF) avoid the left lateral decubitus (LLD) position during sleep and, if so, whether this avoidance would be more pronounced in those with greater degrees of cardiomegaly. BACKGROUND: Anecdotal reports suggest that, in patients with CHF, the LLD position is associated with discomfort due to the enlarged apical heart beat and greater degree of dyspnea (trepopnea) than other positions. It has also been suggested that the LLD position is associated with increased sympathetic nervous activity. METHODS: A total of 75 patients with CHF and 75 control subjects underwent nocturnal polysomnography with monitoring of body position. Echocardiography was performed in all patients with CHF to determine left ventricular end-diastolic diameter (LVEDD). A total of 40 patients underwent cardiac catheterization from which pulmonary capillary wedge pressure (PCWP) and cardiac output (CO) were obtained. RESULTS: Patients with CHF spent significantly less time in the LLD position than in the right lateral decubitus position. No such difference was observed among control subjects. Among patients with CHF, those with larger LVEDD, higher PCWP, and lower CO spent less time in the LLD position. CONCLUSIONS: Patients with CHF avoid the LLD position spontaneously during sleep. This may be a protective strategy to avoid discomfort from the enlarged apical heart beat or further hemodynamic or autonomic compromise. PMID- 12535815 TI - Comparison of the prognostic value of cardiac iodine-123 metaiodobenzylguanidine imaging and heart rate variability in patients with chronic heart failure: a prospective study. AB - OBJECTIVES: We sought to prospectively compare the prognostic value of cardiac iodine-123 (I-123) metaiodobenzylguanidine (MIBG) imaging with that of heart rate variability (HRV) in patients with mild-to-moderate chronic heart failure (HF). BACKGROUND: Cardiac I-123 MIBG imaging, which reflects cardiac adrenergic nerve activity, provides prognostic information on chronic HF patients. Reduced HRV, indicating derangement in cardiac autonomic control, was also reported to be associated with a poor prognosis in chronic HF patients. METHODS: At study entry, I-123 MIBG imaging and 24-h Holter monitoring were performed in 65 chronic HF outpatients with a radionuclide left ventricular ejection fraction <40%. The cardiac MIBG heart to mediastinum ratio (H/M) and washout rate (WR) were obtained from MIBG imaging. The time and frequency domain parameters of HRV were calculated from 24-h Holter recordings. RESULTS: At a mean follow-up of 34 +/- 19 months, WR (p < 0.0001), H/M on the delayed image (p = 0.01), and normalized very low-frequency power (n-VLFP) (p = 0.047) showed a significant association with the cardiac events (sudden death in 3 and hospitalization for worsening chronic HF in 10 patients) on univariate analysis. Multivariate analysis revealed that WR was the only independent predictor of cardiac events, although the predictive accuracy for the combination of abnormal WR and n-VLFP significantly increased, compared with that for abnormal WR (82% vs. 66%, p < 0.05). CONCLUSIONS: Cardiac MIBG WR has a higher prognostic value than HRV parameters in patients with chronic HF. The combination of abnormal WR and n-VLFP would be useful to identify chronic HF patients at a higher risk of cardiac events. PMID- 12535816 TI - Usefulness of invasive electrophysiologic testing to stratify the risk of arrhythmic events in asymptomatic patients with Wolff-Parkinson-White pattern: results from a large prospective long-term follow-up study. AB - OBJECTIVES: The aim of this study was to assess in a large cohort of asymptomatic subjects with Wolff-Parkinson-White (WPW) pattern the usefulness of invasive electrophysiologic testing (EPT) in predicting the occurrence of arrhythmic events over a five-year follow-up. BACKGROUND: Sudden death may be the first clinical manifestation of the WPW syndrome in previously asymptomatic patients. Serial EPTs have been proposed to identify patients at risk. METHODS: A total of 212 consecutive asymptomatic WPW patients were enrolled after a baseline EPT; patients were followed for five years, and 162 patients (115 noninducible and 47 inducible) patients underwent a second EPT. RESULTS: After a mean follow-up of 37.7 months, 33 patients became symptomatic. Of the 115 noninducible patients, 18.2% lost anterograde accessory pathway (AP) conduction, 30% retrograde AP conduction, and only 4 (3.4%) developed symptomatic supraventricular tachycardia (SVT). Of the 47 inducible patients, 25 with sustained atrioventricular reciprocating tachycardia (AVRT) and atrial fibrillation (AF), and 4 with nonsustained AVRT and AF became symptomatic for SVT (n = 21) and AF (n = 8). They were younger, had shorter AP anterograde refractory periods, and multiple APs compared to patients who remained asymptomatic (for all comparisons, p < 0.0001). Of the eight patients with symptomatic episodes of AF and inducible sustained AF, two had a resuscitated cardiac arrest and one died suddenly; all three patients were inducible for AVRT and AF and had multiple APs. CONCLUSIONS: In asymptomatic WPW subjects, EPT may be a valuable tool to stratify the risk of symptomatic and fatal arrhythmic events. PMID- 12535817 TI - Asymptomatic Wolff-Parkinson-White syndrome: is it time to revisit guidelines? PMID- 12535818 TI - Amiodarone and the risk of bradyarrhythmia requiring permanent pacemaker in elderly patients with atrial fibrillation and prior myocardial infarction. AB - OBJECTIVES: The aim of this study was to determine whether the use of amiodarone in patients with atrial fibrillation (AF) increases the risk of bradyarrhythmia requiring a permanent pacemaker. BACKGROUND: Reports of severe bradyarrhythmia during amiodarone therapy are infrequent and limited to studies assessing the therapy's use in the management of patients with ventricular arrhythmias. METHODS: A study cohort of 8,770 patients age > or =65 years with a new diagnosis of AF was identified from a provincewide database of Quebec residents with a myocardial infarction (MI) between 1991 and 1999. Using a nested case-control design, 477 cases of bradyarrhythmia requiring a permanent pacemaker were matched (1:4) to 1,908 controls. Multivariable logistic regression was used to estimate the odds ratio (OR) of pacemaker insertion associated with amiodarone use, controlling for baseline risk factors and exposure to sotalol, Class I antiarrhythmic agents, beta-blockers, calcium channel blockers, and digoxin. RESULTS: amiodarone use was associated with an increased risk of pacemaker insertion (OR: 2.14, 95% confidence interval [CI]: 1.30 to 3.54). This effect was modified by gender, with a greater risk in women versus men (OR: 3.86, 95% CI: 1.70 to 8.75 vs. OR: 1.52, 95% CI: 0.80 to 2.89). Digoxin was the only other medication associated with an increased risk of pacemaker insertion (OR: 1.78, 95% CI: 1.37 to 2.31). CONCLUSIONS: This study suggests that the use of amiodarone in elderly patients with AF and a previous MI increases the risk of bradyarrhythmia requiring a permanent pacemaker. The finding of an augmented risk of pacemaker insertion in elderly women receiving amiodarone requires further investigation. PMID- 12535819 TI - Amiodarone versus placebo and class Ic drugs for cardioversion of recent-onset atrial fibrillation: a meta-analysis. AB - OBJECTIVES: This meta-analysis compared amiodarone with placebo and class Ic drugs for the cardioversion of recent-onset atrial fibrillation (AF), defined as lasting less than seven days. BACKGROUND: Despite the lack of trials that support its efficacy convincingly, amiodarone is widely used for conversion of recent onset AF. METHODS: We searched Medline and EMBASE databases, as well as the Cochrane Controlled Trials Register for randomized trials on recent-onset AF comparing amiodarone to placebo or class Ic drugs. Data were combined according to a fixed effect model. The primary end point was the rate of conversion at 24 h. To study time-dependency of the drugs, efficacy at 1 to 2 h, 3 to 5 h, 6 to 8 h, and at 24 h was analyzed. RESULTS: We found six studies randomizing amiodarone versus placebo (595 patients) and seven studies versus class Ic drugs (579 patients). There was no significant difference between amiodarone and placebo at 1 to 2 h, but significant efficacy was found after 6 to 8 h (relative risk [RR] 1.23, p = 0.022) and at 24 h (RR 1.44, p < 0.001). Efficacy with amiodarone was inferior to class Ic drugs for up to 8 h (RR 0.67, p < 0.001) but no difference was seen at 24 h (RR 0.95, p = 0.50). There were no major adverse effects. CONCLUSIONS: Amiodarone is superior to placebo for cardioversion of AF, and even though the onset of conversion is delayed, its efficacy is similar at 24 h compared with class Ic drugs. These results favor amiodarone as a reasonable alternative for patients with recent AF in whom class Ic and other, more rapidly acting antiarrhythmic drugs cannot be used. PMID- 12535820 TI - Combined intense lifestyle and pharmacologic lipid treatment further reduce coronary events and myocardial perfusion abnormalities compared with usual-care cholesterol-lowering drugs in coronary artery disease. AB - OBJECTIVES: The purpose of this study was to determine if combined intense lifestyle and pharmacologic lipid treatment reduce myocardial perfusion abnormalities and coronary events in comparison to usual-care cholesterol lowering drugs and whether perfusion changes predict outcomes. BACKGROUND: Lifestyle and lipid drugs separately benefit patients with coronary artery disease (CAD). METHODS: A total of 409 patients with CAD, who underwent myocardial perfusion imaging by dipyridamole positron emission tomography at baseline and after 2.6 years, had quantitative size/severity of perfusion defects measured objectively by automated software with follow-up for five additional years for coronary artery bypass graft, percutaneous coronary intervention, myocardial infarction, or cardiac death. Patients were categorized blindly according to prospective, predefined criteria as "poor" treatment without diet or lipid drugs, or smoking; "moderate" treatment on American Heart Association diet and lipid-lowering drugs or on strict low-fat diet (<10% of calories) without lipid drugs; and "maximal" treatment with diet <10% of calories as fat, regular exercise, and lipid active drugs dosed to target goals of low-density lipoproteins <2.3 mmol/l (90 mg/dl), high-density lipoproteins >1.2 mmol/l (45 mg/dl), and triglycerides <1.1 mmol/l (100 mg/dl). RESULTS: Over five years, coronary events occurred in 6.6%, 20.3%, and 30.6% of patients on maximal, moderate, and poor treatment, respectively (p = 0.001). Size/severity of perfusion abnormalities significantly decreased for patients receiving maximal treatment and increased for patients undergoing moderate and poor treatment (p = 0.003 and 0.0001, respectively). Combined intense lifestyle change plus lipid active drugs and severity/change of perfusion abnormalities independently predicted cardiac events. CONCLUSIONS: Intense lifestyle and pharmacologic lipid treatment reduce size/severity of myocardial perfusion abnormalities and cardiac events compared with usual-care cholesterol-lowering drugs. Perfusion changes parallel treatment intensity and predict outcomes. PMID- 12535821 TI - In the statin era, how important are intense lifestyle changes? PMID- 12535822 TI - Raised serum levels of soluble CD40 ligand in patients with familial hypercholesterolemia: downregulatory effect of statin therapy. AB - OBJECTIVES: In the present study, we investigated the effects of statins on serum levels of soluble CD40 ligand (sCD40L) in patients with familial hypercholesterolemia (FH). BACKGROUND: Atherosclerotic disease seems to involve inflammatory and immunologic mechanisms, and sCD40L has recently been identified as one of the key players in the atherosclerotic process. HMG-Co A reductase inhibitors, statins, have been recognized as immunomodulators and reduce cardiovascular events and mortality, but the effects of statins on sCD40L has not been clarified. METHODS: In a randomized, double-blind, clinical trial, as part of the Atorvastatin versus Simvastatin on Atherosclerosis Progression (ASAP) trial, 110 patients with FH were given atorvastatin 80 mg/daily (n = 57) or simvastatin 40 mg/daily (n = 53) for two years. RESULTS: Our main findings were: 1) at baseline patients with FH had significantly higher (approximately 27-fold) serum levels of sCD40L than healthy controls; 2) statin therapy markedly decreased serum levels of sCD40L (approximately 40% reduction); 3) this decrease in sCD40L was found during both "aggressive" (i.e., atorvastatin) and "conventional" (i.e., simvastatin) statin therapy and was not correlated with the degree of reduction in cholesterol levels. CONCLUSIONS: Our findings may suggest enhanced CD40L-CD40 interaction in FH and that this inflammatory response may be downregulated by statins. PMID- 12535823 TI - Echocardiographic characteristics of professional football players. AB - OBJECTIVES: We examined the echocardiographic characteristics of highly trained American football players. BACKGROUND: Intense physical training is associated with morphologic and physiologic cardiac changes often referred to as the "athlete's heart." Echocardiographic features peculiar to elite football players have not been described. METHODS: We studied cardiac morphology and function as assessed by rest and stress echocardiography in 156 asymptomatic National Football League players. Resting and stress ejection fraction (EF), wall thickness, and diastolic left ventricular internal diameter (LVID) were measured. Left ventricular (LV) mass was calculated, as was relative wall thickness (RWT) defined as septal and posterior wall thickness divided by LVID. Control data were obtained from published studies. RESULTS: The mean LVID (53 +/- 0.5 mm) and maximal wall thickness (11.2 +/- 0.2 mm) were increased over normal reported control subjects. There was a correlation between LVID and body weight (p = 0.01) and body surface area (BSA) (p = 0.01). The average LVID indexed to BSA was 23 +/ 2 mm/M(2). There was also a correlation between maximal wall thickness and body weight (p = 0.01) and BSA (p = 0.01). The average wall thickness indexed to BSA was 5.05 +/- 0.88 mm/M(2). Of the players, 23% had evidence of LV hypertrophy. Two players had an increased septal-to-posterior-wall-thickness ratio (> or =1.3), although no player had an outflow gradient. The RWT for the players was 0.424 (+/- 0.1). The mean resting EF was 58% (+/- 4.4%), and every player undergoing exercise testing had an appropriate hyperdynamic response in cardiac function. CONCLUSIONS: Both wall thickness and LVID of elite American football players are increased and correlate with body size. There is a high RWT, reflecting an emphasis on strength training. The LV EF was normal and not supranormal, as is sometimes believed. Regardless of the resting EF, all players had hyperdynamic cardiac responses with exercise. PMID- 12535824 TI - Intracardiac echocardiography guided device closure of atrial septal defects. AB - OBJECTIVES: This study was designed to determine the feasibility and accuracy of intracardiac echocardiography (ICE) in guiding percutaneous closure of atrial septal defects (ASD). BACKGROUND: Intracardiac echocardiography is a novel imaging technique that might be used to guide interventional procedures. The sensitivity and specificity of ICE, compared to standard imaging techniques, in detecting potentially adverse procedural events and guiding remedial action will be an important consideration in its use. METHODS: In a prospective study, 24 patients underwent device closure of ASD using ICE as the primary echocardiographic imaging modality. Feasibility was expressed as proportion of cases in which complete diagnostic ICE imaging was achieved. Accuracy was expressed as the percent agreement between ICE and simultaneously performed transesophageal echocardiography (TEE). RESULTS: High-quality ICE images were acquired in all patients, though images were limited in two patients with aneurysmal septa. Intracardiac echocardiography successfully guided closure of 24 out of 25 ASDs (96%) in 23 patients. There was close agreement between ICE and TEE in their assessment of device position and the adequacy of septal capture before device release (98%) and in identifying the presence of significant residual shunts. Intracardiac echocardiography detected all potentially adverse events, including four malpositions, and guided appropriate remedial action. CONCLUSIONS: Intracardiac echocardiography guided device closure of secundum ASDs is feasible in the majority of patients and provides diagnostic data comparable to TEE. These data indicate that ICE may be used to guide routine closure of ASDs in adults without the need for TEE and general anesthesia. PMID- 12535825 TI - Impact of aging on substrate metabolism by the human heart. AB - BACKGROUND: Results of studies in experimental animals have shown that, with age, myocardial fatty acid metabolism decreases, and glucose metabolism increases. Whether similar changes occur in humans is unknown. METHODS: Seventeen healthy younger normal volunteers (six males, 26 +/- 5 years) and 19 healthy older volunteers (nine males, 67 +/- 5 years) underwent positron emission tomography (PET) under resting conditions in the fasted state. Myocardial blood flow (MBF), myocardial oxygen consumption (MVO(2)), myocardial fatty acid utilization (MFAU) and oxidation (MFAO), and myocardial glucose utilization (MGU) were quantified by PET with (15)O-water, (11)C-acetate, (11)C-palmitate, and(11)C-glucose, respectively. RESULTS: Although MBF was similar between the groups, MVO(2) was higher in the older subjects (5.6 +/- 1.6 micromol/g/min) compared with younger subjects (4.6 +/- 1.0 micromol/g/min, p < 0.04). Rates of MFAU and MFAO (corrected for MVO(2)) were significantly lower in older subjects than in younger subjects (MFAU/MVO(2): 35 +/- 10 vs. 51 +/- 20 nmol free fatty acids (FFA)/nmol O(2) x 10(-3), p < 0.005, and MFAO/MVO(2): 33 +/- 10 vs. 48 +/- 18 nmol FFA/nmol O(2) x 10(-3), p < 0.004). In contrast, the rates of MGU corrected for MVO(2) did not differ between the groups. CONCLUSIONS: With aging, humans exhibit a decline in MFAU and MFAO. Although absolute rates of MGU do not increase, by virtue of the decline in MFAU there is likely an increase in relative contribution of MGU to substrate metabolism. The clinical significance of this metabolic switch awaits further study. PMID- 12535826 TI - Gender differences in post-infarction hypertrophy in end-stage failing hearts. AB - OBJECTIVES: We explored whether there are gender differences in cardiac remodeling and whether etiology influences organ and cellular remodeling in advanced heart failure (HF). BACKGROUND: Several studies have shown a survival benefit for women compared to men with symptomatic HF. This observation may be related to gender differences in cardiac remodeling. METHODS: We studied hearts from 100 patients (72 men and 28 women) receiving cardiac transplantation at our institution. Cardiac morphology was assessed with echocardiography and direct measurement of cardiac mass. Cardiac myocyte volume, length, width, cross sectional area, and contraction were measured using previously validated techniques. RESULTS: Among 50 patients with idiopathic cardiomyopathy (CM), we observed no gender-based differences in cardiac or cellular remodeling. In contrast, among 50 patients with ischemic cardiomyopathy (ICM), the heart weight index was significantly greater in men, and there was a strong trend toward an increased left ventricular (LV) mass index as well. These gender differences in cardiac and LV mass were paralleled by marked gender differences in myocyte volume, such that average myocyte volume was 36% greater in men than in women, in association with a 14% increase in resting cell length. CONCLUSIONS: Our studies demonstrate a multilevel gender difference in post-infarction remodeling, with women exhibiting reduced hypertrophy. Our studies further demonstrate that gender differences in cardiac remodeling in ICM are largely related to fundamental differences in cellular remodeling rather than simply differences in infarct size or expansion. Distinctions observed between ischemic and idiopathic CM suggest that gender may influence local myocardial responses to injury. PMID- 12535827 TI - Gender differences in recovery after coronary artery bypass surgery. AB - OBJECTIVES: This study was designed to examine whether female gender is associated with poorer recovery after coronary artery bypass graft (CABG) surgery. BACKGROUND: The risks and benefits associated with CABG surgery in women are not as well established as they are in men, and there are concerns that women may have worse outcomes. The recovery period after CABG (the first four to eight weeks after the surgery) is a vulnerable time, with higher risks of complications and hospital readmission. There is little information on patients' experiences during this phase, particularly among women. METHODS: We prospectively followed 1,113 patients (804 men and 309 women) who underwent first CABG consecutively between February 1999 and February 2001. Patients were interviewed at baseline and between six and eight weeks after surgery. Clinical data were abstracted from medical records. RESULTS: Compared with men, women were older and more often had unstable angina and congestive heart failure, lower physical function (PF), and more depressive symptoms in the month before surgery. At six to eight weeks after CABG surgery, after adjustment for baseline characteristics, the rate of hospital readmission was 20.5% in women and 11.0% in men (p = 0.005), and the mean number of physical symptoms and side effects was 2.5 in women and 2 in men (p = 0.0009). Whereas, on average, PF remained unchanged in men (an increase in score of 0.3 points, 95% confidence interval [CI], -1.1 to 1.8) and depressive symptoms improved (a decrease of 0.2 depressive symptoms, 95% CI, -0.4 to -0.04), women showed, on average, a 13-point decline in physical function (95% CI, -15.8 to 10.4) and an increase of 0.5 in depressive symptoms (95% CI, 0.1 to 0.9). CONCLUSIONS: After CABG surgery, women have a more difficult recovery compared with men, which is not explained by illness severity, presurgery health status, or other patient characteristics. PMID- 12535828 TI - Maximum left ventricular thickness and risk of sudden death in patients with hypertrophic cardiomyopathy. AB - OBJECTIVES: We sought to assess the relationship between maximum left ventricular (LV) wall thickness and outcome in patients with hypertrophic cardiomyopathy (HCM). BACKGROUND: An association between maximum LV wall thickness and risk of sudden death was suggested in HCM. This finding requires further investigation, given the important implications for risk stratification and treatment. METHODS: We analyzed the mortality and risk profile of 237 patients (age 41 +/- 17 years; 63% male) classified into five groups based on echocardiographic maximum LV thickness. RESULTS: During follow-up (12 +/- 7 years), 36 patients died of cardiovascular causes, including 16 sudden deaths. Maximum LV thickness was not associated with a risk of sudden death (p = 0.37) nor with overall cardiovascular mortality (p = 0.7). With the exception of the small subset with thickness values < or =15 mm, with a consistently benign clinical course, the distribution of sudden death and overall cardiovascular mortality was not significantly different among the other four classes, ranging from 16 to 19 mm to > or =30 mm. Among 30 patients with extreme LV thickness (> or =30 mm), only one sudden event occurred among six patients diagnosed at <18 years of age (17%) and none among 24 diagnosed at > or =18 years of age. The prevalence of nonsustained ventricular tachycardia, syncope, an abnormal blood pressure response to exercise, and atrial fibrillation was similar among the five thickness classes. CONCLUSIONS: During 12 year follow-up, we observed no association between maximum LV thickness and cardiovascular mortality in a community-based population with HCM. The degree of maximum LV wall thickness should be considered in the context of a multifactorial approach to risk stratification, rather than as an isolated risk factor. Only in those patients diagnosed at a very young age might the presence of extreme LV wall thickness represent, per se, a potential marker of risk of sudden death. PMID- 12535829 TI - A clinical and histopathologic comparison of cardiac sarcoidosis and idiopathic giant cell myocarditis. AB - OBJECTIVES: The goal of this study was to determine the prognostic value of clinical data available at presentation and histology in cardiac sarcoidosis (CS) and idiopathic giant cell myocarditis (IGCM). BACKGROUND: The prognosis of patients with nonischemic cardiomyopathy is partly dependent on the histologic diagnosis. Survival in IGCM is poor. The prognosis of a histologically related entity, cardiac sarcoidosis (CS), is less well established, and the prognostic value of the distinction between CS and IGCM on endomyocardial biopsy (EMB) is unknown. METHODS: We identified 115 patients from the Multicenter IGCM Registry with CS (n = 42) and IGCM (n = 73). We compared the clinical data for these two groups using Cox proportional-hazards models to assess the association between histologic diagnosis and survival. In order to determine whether histologic features could reliably differentiate these two entities, two cardiac pathologists semiquantitatively graded the inflammatory infiltrate components and compared the results between groups. RESULTS: Black race was more frequent in the CS group (31% vs. 4%, p < 0.0001). Syncope and atrioventricular block were also more frequently observed in CS than IGCM (31% vs. 5%, p = 0.0002 and 50% vs. 15%, p < 0.0001, respectively). Left-sided heart failure was more common in IGCM (40% vs. 64%, p = 0.013). In CS patients diagnosed by EMB, the five-year transplant free survival after diagnosis was 69.8% versus 21.9% for IGCM (p < 0.0001, log rank test). In multivariate models, presentation with heart failure predicted IGCM, and presentation with heart block or more than nine weeks of symptoms predicted CS. Eosinophils, myocyte damage, and foci of lymphocytic myocarditis were more frequent in IGCM, while granulomas and fibrosis were more frequent in CS. CONCLUSIONS: Transplant-free survival is better for patients with CS than for IGCM diagnosed by EMB. Presentation with heart failure predicted IGCM, and presentation with heart block or more than nine weeks of symptoms predicted CS. PMID- 12535830 TI - Long-term outcome in patients with Marfan syndrome: is aortic dissection the only cause of sudden death? AB - OBJECTIVES: We sought to assess outcomes in a series of young patients with Marfan syndrome and to define the prevalence of ventricular arrhythmias in this patient population. BACKGROUND: While sudden death is a well-recognized outcome in Marfan syndrome, ventricular arrhythmias are not well described. METHODS: Patients were followed with echocardiography, electrocardiography, and ambulatory electrocardiography. The prevalence and associated factors for ventricular dysrhythmias were defined. RESULTS: Seventy patients with Marfan syndrome diagnosed at birth to 52 years were followed for a period of up to 24 years. All patients had cardiovascular involvement and were started on medical therapy. No patient died from aortic dissection, while 4% died from arrhythmias. Ventricular arrhythmias were present in 21% and were associated with increased left ventricular size, mitral valve prolapse, and abnormalities of repolarization. CONCLUSIONS: Cardiac complications are rare in young patients with Marfan syndrome receiving medical therapy and close clinical follow-up. Sudden death still occurs, and appears more common in patients with a dilated left ventricle. Left ventricular dilation may predispose to alterations of repolarization and fatal ventricular arrhythmias. PMID- 12535831 TI - Acute systemic inflammation enhances endothelium-dependent tissue plasminogen activator release in men. AB - OBJECTIVES: The purpose of this study was to investigate in vivo the effects of acute systemic inflammation on the endogenous fibrinolytic capacity in men. BACKGROUND: Systemic inflammation and endogenous fibrinolysis play a major role in the pathogenesis of coronary artery disease. Although previous studies have shown impaired endothelium-dependent vasomotor function, the effects of inflammation on the endothelial release of the fibrinolytic factor tissue plasminogen activator (t-PA) are unknown. METHODS: In a double-blind randomized placebo-controlled crossover trial, we administered a mild inflammatory stimulus, Salmonella typhi vaccine, or saline placebo to eight healthy men on two separate occasions. Six hours after vaccination, blood flow and plasma fibrinolytic variables were measured in both arms during intrabrachial infusions of bradykinin (40 to 1,000 pmol/min), acetylcholine (5 to 20 microg/min), and sodium nitroprusside (2 to 8 microg/min). RESULTS: Compared with placebo, the S. typhi vaccination caused a rise in white cell count (11.1 +/- 0.5 x10(9)/l vs. 7.9 +/- 0.8 x10(9)/l; p = 0.004) and plasma interleukin-6 concentrations (6.9 +/- 1.4 pg/ml vs. 1.6 +/- 0.4 pg/ml; p = 0.01) in addition to a significant augmentation of t-PA antigen (45 +/- 9 ng/100 ml/min at peak dose vs. 24 +/- 8 ng/100 ml/min at peak dose; p = 0.016, analysis of variance) and activity (104 +/- 15 IU/100 ml/min vs. 54 +/- 12 IU/100 ml/min; p = 0.006, analysis of variance) release during bradykinin infusion. Forearm blood flow increased in a dose-dependent manner after bradykinin, acetylcholine and sodium nitroprusside infusions (p < 0.001), but this was unaffected by vaccination. CONCLUSIONS: Our results showed that acute systemic inflammation augmented local forearm t-PA release in men, which suggests that acute inflammation may invoke a protective response by enhancing the acute endogenous fibrinolytic capacity in healthy men. Further studies are needed to clarify whether this response is impaired in patients with cardiovascular disease. PMID- 12535832 TI - Carvedilol increases the production of interleukin-12 and interferon-gamma and improves the survival of mice infected with the encephalomyocarditis virus. AB - OBJECTIVES: This study was designed to examine the effects of carvedilol in a murine model of viral myocarditis induced by encephalomyocarditis virus (EMCV) infection. BACKGROUND: Cytokines play an important role in the pathophysiology of viral myocarditis. Catecholamines influence the production of cytokines via beta adrenergic receptors, suggesting that beta-adrenergic blockers could modulate the production of cytokines and exert a therapeutic effect in viral myocarditis by blocking the beta-stimulating action of endogenous catecholamines. In clinical trials, the third-generation, nonselective beta-blocker carvedilol was the first among several beta-blockers to reduce mortality in heart failure. However, the effects of carvedilol in acute viral myocarditis and on cytokine production are unknown. METHODS: This study compared the effects of carvedilol, the selective beta(1)-blocker metoprolol, and the nonselective beta-blocker propranolol in a murine model of viral myocarditis induced by EMCV. RESULTS: Carvedilol improved the 14-day survival of the animals, attenuated myocardial lesions on day 7, and increased myocardial levels of interleukin (IL)-12 and interferon (IFN)-gamma, whereas reducing myocardial virus replication. Propranolol also attenuated myocardial lesions, but to a lesser extent, and increased IL-12 and IFN-gamma levels. Metoprolol had no effect in this model. Encephalomyocarditis virus infection increased plasma catecholamine levels. CONCLUSIONS: These results suggest that by blocking the beta(2)-stimulating effects of catecholamines, carvedilol exerts some of its beneficial effects by increasing the production of IL-12 and IFN-gamma. Carvedilol may be effective in patients with viral myocarditis by boosting IL-12 and IFN-gamma production. PMID- 12535833 TI - President's page: medical history: a valuable tool to help us frame the present and predict the future. PMID- 12535834 TI - Chromatographic performance of large-pore versus small-pore columns in micellar liquid chromatography. AB - Micellar liquid chromatography (MLC) is useful in bioanalysis because proteinaceous biofluids can be directly injected onto the column. The technique has been limited in part because of the apparently weak eluting power of micellar mobile phases. It has recently been shown [Anal. Chem. 72 (2000) 294] that this may be overcome by the use of large pore size stationary phases. In this work, large-pore (1000 A) C(18) stationary phases were evaluated relative to conventional small-pore (100 A) C(18) stationary phases for the direct sample injection of drugs in plasma. Furthermore, the difference between the large and small pore phases in gradient elution separations of mixtures of widely varying hydrophobicities was investigated. Large-pore stationary phases were found to be very effective for eluting moderately to highly hydrophobic compounds such as ibuprofen, crotamiton, propranolol, and dodecanophenone, which were highly retained on the small-pore stationary phases typically used in MLC. The advantages of direct introduction of biological samples (drugs in plasma) and rapid column re-equilibration after gradient elution in MLC were maintained with large-pore phases. Finally, recoveries, precision, linearity, and detection limits for the determination of quinidine and DPC 961 in spiked bovine plasma were somewhat better using MLC with wide pore phases. PMID- 12535835 TI - Alkylation of DNA by melphalan: investigation of capillary liquid chromatography electrospray ionization tandem mass spectrometry in the study of the adducts at the nucleoside level. AB - Nitrogen mustards are among the oldest cancer chemotherapeutic agents and remain the drugs of choice for treatment of many human cancers. A serious complication of treatment with nitrogen mustards is the increased risk of a secondary leukaemia in long-term survivors because not all alkylating agent interactions with DNA result in cell death. In an earlier study 2'-deoxy-5' mononucleotide/melphalan adducts have been analysed by us by LC-ES MSMS. In this work we want to present the first results of the analysis of the corresponding 2' deoxynucleoside/melphalan adducts from DNA hydrolysates by column switching/capillary LC-ES tandem mass spectrometry. Nucleosides, compared to nucleotides, give better chromatographic results and show a good sensitivity under electrospray (+) [ES(+)] ionisation. Several adducts were identified under ES(+) conditions. Mono-alkylated nucleoside adducts alkylated at the base moiety were identified for dGuo, dCyd and dAdo. Structures were identified by recording the low-energy CAD product ion scans. Also a mono-alkylated nucleotide pdA with alkylation position at the phosphate moiety could be detected. This proves that in the case of phosphate alkylation the enzymatic dephosphorylation reaction was inhibited. A Jurkat cell suspension was treated with melphalan (1 mM) and incubated at 37 degrees C (5% CO(2)). After 6 and 48 h, the DNA was isolated and enzymatically hydrolysed. The corresponding nucleoside pool was evaluated with the developed LC-MS method. In the 48-h experiment, one adduct could be identified as a N-7 alkylated dGuo. In the 6-h experiment, no adducts could be found. Additional experiments were done wherein Jurkat-DNA, isolated from a non treated cell culture, was treated with melphalan. These results were analogous with the data found in melphalan-treated calf thymus DNA. Additionally, we tried to determine the exact alkylation position by interpreting high-resolution fragmentation spectra. PMID- 12535836 TI - Determination of riboflavin in urine by capillary electrophoresis-blue light emitting diode-induced fluorescence detection combined with a stacking technique. AB - A simple, inexpensive and reliable method for the simultaneous, routine analysis of riboflavin in urine by capillary electrophoresis-light emitting diode (LED) induced fluorescence detection is described. Using a blue LED as the light source, the detection limit of riboflavin was determined to be 0.48 microg/ml and was improved to 20 ng/ml when a stacking technique was applied. In the analysis of an actual sample, various concentrations of riboflavin were distributed in the urine samples over a period of 9 h after the ingestion of a vitamin B(2) tablet. PMID- 12535837 TI - Liquid chromatography-tandem mass spectrometry analysis of oleuropein and its metabolite hydroxytyrosol in rat plasma and urine after oral administration. AB - We describe a liquid chromatography-electrospray ionisation tandem mass spectrometry method for the qualitative and quantitative determination of the secoiridoid oleuropein and its bioactive metabolite hydroxytyrosol in rat plasma and urine. Samples were prepared by liquid-liquid extraction using ethyl acetate with a recovery for both compounds of about 100% in plasma and about 60% in urine. The chromatographic separation was performed with a RP-ODS column using a water-acetonitrile linear gradient. The calibration curve was linear for both biophenols over the range 2.5-1000 ng/ml (LOD 1.25 ng/ml) for plasma and 5-1000 ng/ml (LOD 2.5 ng/ml) for urine. Plasma concentrations of oleuropein and hydroxytyrosol were measured after oral administration of a single dose (100 mg/kg) of oleuropein. Analysis of treated rat plasma showed the presence of unmodified oleuropein, reaching a peak value of 200 ng/ml within 2 h, with a small amount of hydroxytyrosol, whereas in urine, both compounds were mainly found as glucuronides. PMID- 12535838 TI - Determination of uric acid in human saliva by high-performance liquid chromatography with amperometric electrochemical detection. AB - The aim of the present study is to establish a highly sensitive method for the determination of uric acid (UA) in human saliva. The monitoring of UA levels in less invasive biological samples such as saliva is suggested for the diagnosis and therapy of gout, hyperuricemia, and the Lesch-Nyhan syndrome, and for detecting such conditions as alcohol dependence, obesity, diabetes, high cholesterol, high blood pressure, kidney disease, and heart disease. Reversed phase high-performance liquid chromatography with electrochemical detection (HPLC ED) was employed for the determination of UA obtained by solid-phase extraction from saliva. To quantify UA, we compared the ED efficiencies of an amperometric ED (Ampero-ED) with a single electrode and a coulometric ED (Coulo-ED) with a multiple electrode array. The results showed that the detection limits (S/N=3) were 3 nM for Ampero-ED and 6 nM for Coulo-ED, and the linearity of the calibration curves of 60-6000 nM had correlation coefficients exceeding 0.999. In addition, the total analytical time was 10 min. In the sample preparation of UA in saliva, an Oasis MAX solid-phase cartridge was used. The recoveries of UA spiked at 0.6 and 3 microM in saliva were above 95% with a relative standard deviation (RSD) of less than 15%. Therefore, the present method may be used in the routine and diagnostic determination of UA in human saliva. PMID- 12535839 TI - Measurement of the anti-cancer agent gemcitabine in human plasma by high performance liquid chromatography. AB - A reversed-phase HPLC assay has been developed to determine the concentration of the anti-metabolite 2',2'-difluorodeoxycytidine (gemcitabine, dFdC) in human plasma over the concentration range of 0.5-150 microM (0.13-39.44 microg/ml), and 2',2'-difluorodeoxyuridine (dFdU), the deaminated, inactive metabolite, over the range of 1.0-227 microM (0.26-60 microg/ml). After the addition of 20 nmol 2' fluorodeoxycytidine (FdC) as an internal standard, 0.5-ml samples of plasma were subjected to acetonitrile precipitation, followed by analysis using a gradient reversed-phase HPLC assay with UV detection. A Phenomenex Columbus C(18) column, 5 microm, 150 x 4.6 mm, and a Waters C(18), 4 microm, Nova-Pak Sentry guard column were used to achieve separation. FdC, dFdC and dFdU were monitored at 282, 269 and 258 nm, respectively, on a Waters 996 photodiode array detector. The mobile phase, run at a total flow-rate of 1.5 ml/min, was composed of two solvents: 50 mM ammonium acetate pH 5.0 in either 2% (solvent A) or 10% methanol (solvent B, v/v); 100% solvent A was run for 17 min, followed by a linear gradient to 100% solvent B over 14 min. FdC, dFdC and dFdU were resolved from endogenous compounds and had retention times of 13.6+/-0.5, 18.1+/-1.1 and 29.0+/ 0.6 min, respectively. The assay was useful in measuring the plasma levels of both analytes in samples obtained from adult cancer patients participating in a Phase I trial of gemcitabine given as either a 1- or 2-h infusion weekly for 3 of 4 weeks. PMID- 12535840 TI - Validation of a simple gas chromatographic-mass spectrometric method for the determination of gamma-butyrolactone in human plasma. AB - A gas chromatographic-mass spectrometric (GC-MS) method is described for the determination of human plasma levels of gamma-butyrolactone (GBL) is described. The method is sensitive and simple. The plasma sample spiked with the internal standard was extracted by dichloromethane (CH(2)Cl(2)) in acidic conditions, and the concentrated organic layer was injected into GC-MS. Because of endogenous GBL in human plasma, the method used a standard calibration curve. The calibration curve was linear from 10 to 1000 ng/ml. The method has been validated for accuracy and precision with the relative error and C.V. for intra- and inter-day within 10%. GBL-spiked plasma samples stored at -80 degrees C were stable for a 3 month period. The stability of plasma samples after three cycles of freezing and thawing and of prepared samples on an autosampler for 48 h were demonstrated. Plasma concentrations of GBL before and after administration of UFT were 24.3+/ 14.2 and 84.9+/-22.4 ng/ml, respectively. PMID- 12535841 TI - Quantification of the O- and N-demethylated metabolites of hydrocodone and oxycodone in human liver microsomes using liquid chromatography with ultraviolet absorbance detection. AB - High-performance liquid chromatographic assays for the O- and N-demethylated oxidative metabolites of hydrocodone and oxycodone formed in human liver microsomes are described. A solvent-solvent extraction/re-extraction procedure followed by reversed-phase HPLC with UV detection at 210 nm allows for the quantification of hydromorphone, norhydrocodone, oxymorphone and noroxycodone. Calibration curve concentration ranges were 0.63-400 microM (0.18-114 microg/ml) and 1.25-400 microM (0.36-114 microg/ml) for hydromorphone and norhydrocodone, respectively and 0.13-20 microM (0.04-6.03 microg/ml) and 1-200 microM (0.30-60 microg/ml) for oxymorphone and noroxycodone, respectively. Assay performance was determined by intra- and inter-assay precision and inaccuracies for quality control samples and was <15% for all metabolites at each quality control concentration. These methods provide good precision, accuracy and sensitivity for use in in vitro kinetic studies investigating the oxidative metabolism of hydrocodone and oxycodone in human liver microsomes. PMID- 12535842 TI - Simultaneous determination of molecular species of monoacylglycerols, diacylglycerols and triacylglycerols in human very-low-density lipoproteins by reversed-phase liquid chromatography. AB - The aim of the present study was to investigate the applicability of a previously developed method for the analysis of triacylglycerol molecular species to the simultaneous determination of triacylglycerols, diacylglycerols and monoacylglycerols of human very-low-density lipoproteins (VLDL). Ten elderly women were recruited for the study. Blood was obtained in fasting conditions and VLDL were isolated by ultracentrifugation. Neutral lipids were separated by solid phase extraction and were subsequently injected on a reversed-phase HPLC system, with an elution system composed of acetone in acetonitrile. The method allowed the separation of four monoacylglycerols, 18 diacylglycerols and 24 triacylglycerols, including the resolution of positional isomers of diacylglycerols. Monoacylglycerols were composed of oleic, linoleic, palmitic and stearic acids. The major diacylglycerols were 1,2-dilinoleoyl-glycerol and 1,3 dilinoleoyl-glycerol (14.24+/-1.02 and 17.93+/-1.42%, respectively). The main triacylglycerols quantified were dioleoyl-stearoyl-glycerol (OOS), oleoyl dipalmitoyl-glycerol (OPP), trilinoleoyl-glycerol (LLL) and linoleoyl-distearoyl glycerol (LSS), accounting for 11.25+/-2.15, 10.14+/-2.05, 9.35+/-2.30 and 8.56+/ 1.56%, respectively. An inverse relationship between polarity and fatty acid disappearance from triacylglycerols (r(2)=0.82, P<0.05) and from diacylglycerols (r(2)=0.93, P<0.01) was discovered. In conclusion, the method allowed, for the first time, the easy, rapid and simultaneous determination in a single chromatogram of triacylglycerol, diacylglycerol and monoacylglycerol molecular species of human VLDL by reversed-phase HPLC. PMID- 12535843 TI - Improved method for measurement of human plasma xanthine oxidoreductase activity. AB - The XOR activity in human plasma was measured by quantifying the XOR-derived uric acid (UA) in plasma using the high-performance liquid chromatography (HPLC) equipped with a UV detector. Chromatographic separation consisted of the mobile phase (a mixture of 0.1% trifluoroacetic acid in Milli-Q water and 0.085% trifluoroacetic acid in acetonitrile in a mix ratio of 99:1) running through a Zorbax StableBond SB-C(18) column at a flow-rate of 1 ml/min. Deproteinization with heat-treatment of plasma samples after the reaction was used in the assay to avoid splitting of the UA and xanthine peaks caused by acid deproteinization that could interfere the accurate determination of human plasma XOR activity in our case. Based on the examination of the dependence of XOR activity on added amounts of xanthine and reaction times, the amount of xanthine and reaction time for XOR activity assay were determined to prevent the errors caused by the limiting effect of substrates and plateau phase of the reaction. Using this method, human plasma XOR activities of 25 healthy people were measured. The average human plasma XOR activity was 2.1+/-0.8 (x10(-3) U/ml). PMID- 12535844 TI - Sensitive and rapid liquid chromatography-tandem mass spectrometry method for the determination of meloxicam in human plasma. AB - Meloxicam was quantified in human plasma after a single 15 mg oral dose of the drug was given to 26 healthy volunteers. An Applied Biosystems Sciex API 2000 triple quadrupole mass spectrometer in multiple reaction monitoring (MRM) mode, using TurboIonSpray (TIS) in the positive ion mode, was used. Protein precipitation with acetonitrile was followed by C(18) reverse phase liquid chromatography and tandem mass spectrometry. The mean recovery for meloxicam was 92% with a lower limit of quantification of 8.96 ng/ml. Piroxicam was used as the internal standard. This assay method makes use of the increased sensitivity and selectivity of tandem mass spectrometry (MS-MS) detection to allow for a more rapid (extraction and chromatography) and selective method for the determination of meloxicam in human plasma than has previously been described. PMID- 12535845 TI - Determination of valdecoxib and its metabolites in human urine by automated solid phase extraction-liquid chromatography-tandem mass spectrometry. AB - A simple, sensitive and specific automated SPE-LC-MS-MS assay was developed and validated for determination of valdecoxib (I), its hydroxylated metabolite (II) and carboxylic acid metabolite (III) in human urine. The analytes (I, II and III) and a structural analogue internal standard (I.S.) were extracted on a C(18) solid-phase extraction cartridge using a Zymark RapidTrace automation system. The chromatographic separation was performed on a narrow-bore reverse phase HPLC column with a mobile phase of acetonitrile-water (50:50, v/v) containing 10 mM 4 methylmorpholine (pH 6.0). The analytes were ionized using negative electrospray mass spectrometry, then detected by multiple reaction monitoring with a tandem mass spectrometer. The precursor to product ion transitions of m/z 313-->118, m/z 329-->196 and m/z 343-->196 were used to measure I, II and III, respectively. The assay exhibited a linear dynamic range of 1-200 ng/ml for I and II and 2-200 ng/ml for III in human urine. The lower limit of quantitation was 1 ng/ml for I and II and 2 ng/ml for III. Acceptable precision and accuracy were obtained for concentrations over the standard curve ranges. Run time of 5.5 min for each sample made it possible to analyze a throughput of 70 human urine samples per run. The assay has been successfully used to analyze human urine samples to support clinical phase I and II studies. PMID- 12535846 TI - Liquid chromatographic-mass spectrometric determination of cyclooxygenase metabolites of arachidonic acid in cultured cells. AB - A liquid chromatographic-electrospray ionization-mass spectrometric (LC-ESI-MS) technique was developed to simultaneously determine the cyclooxygenase metabolites of arachidonic acid (6-keto-PGF(1alpha), PGD(2), PGE(2), PGF(2alpha), and PGJ(2)) produced by cultured cells. Samples were separated on a C(18) column with water-acetonitrile mobile phase, ionized by electrospray, and detected in the positive mode. Selected ion monitoring (SIM) of m/z 353, 335, 335, 319, and 317 were used for quantifying 6-keto-PGF(1alpha), PGD(2), PGE(2), PGF(2alpha), and PGJ(2), respectively. Prostaglandins were detected at concentrations as low as 1 pg (S/N=3) on the column. The method was used to determine the production of PGs from bovine coronary artery endothelial cells (ECs) and human prostate cancer cells (PC-3) with different degree of invasiveness. Bradykinin (10(-6) M) stimulated a marked increase in the production of 6-keto-PGF(1alpha), PGE(2), and PGF(2alpha) and a small increase of PGD(2) by ECs. 6-Keto-PGF(1alpha) was the major metabolite in these cells. The production of PGE(2) was threefold higher and PGD(2) was twofold higher in PC-3-S (invasive) cells than in PC-3-U (non invasive) cells. PMID- 12535847 TI - Simultaneous determination of free and conjugated bile acids in serum by cyclodextrin-modified micellar electrokinetic chromatography. AB - A simultaneous determination of 15 free and most conjugated forms of bile acids (BA) in serum using capillary electrophoresis is described. The optimized and validated method proposed in this work is straightforward and rapid, employing affordable equipment. A background electrolyte of 5 mM beta-cyclodextrin, 5 mM 2 hydroxypropyl-beta-cyclodextrin, 50 mM SDS and sodium borate-dihydrogen phosphate pH 7.0 with 10% of acetonitrile was used. The complete separation of 15 BA, not easily achievable with other methods, is performed in less than 12 min using a UV detector with good precision and accuracy. BA were extracted from pretreated serum samples using a C(18)-solid-phase extraction and the recovery values ranged from 65 to 107.8%. Limits of quantitation were between 0.58 and 3.2 microM. This method proved to be suitable to determine individual BA profiles which are more useful than total serum bile acids as indicators of metabolic disorders and hepatobiliary diseases. PMID- 12535848 TI - Simultaneous determination of oleuropein and its metabolites in plasma by high performance liquid chromatography. AB - A method based on high-performance liquid chromatography (HPLC) with a diode array detection system was developed and validated aiming at the simultaneous determination of oleuropein (OE) and its metabolites, hydroxytyrosol (HT) and tyrosol (T), in human plasma. These phenolic components are believed to play a vital role in the prevention of coronary artery disease and atherosclerosis. The proposed method includes a clean-up solid-phase extraction procedure (using a C(18) column) with high recovery efficiency (85-100%). The statistical evaluation of the method reveals good linearity, accuracy and reproducibility for all the compounds analyzed with RSD values less than 6.5%, while the detection limit is 50 ng/ml for both OE and T and 75 ng/ml for HT. This assay can be employed in bioavailability studies of olive oil phenolic compounds, thus assisting the evaluation of their pharmacological role. PMID- 12535849 TI - Determination of thalidomide in transport buffer for Caco-2 cell monolayers by high-performance liquid chromatography with ultraviolet detection. AB - We report simple validated HPLC methods for the determination of thalidomide in the transport buffer for the human colonic cell line (Caco-2) cell monolayers. An aliquot of 50 microl of the mixture was injected onto a Spherex C(18) column (150 x 4.6 mm; 5 microm) at a flow-rate of 0.5 ml/min of mobile phase consisting of acetonitrile-10 mM ammonium acetate buffer (24:76, v/v, pH 5.5), and thalidomide was detected by ultraviolet detector at a wavelength of 220 nm. Calibration curves for thalidomide were constructed at the concentration range of 0.025-1.0 and 1.0-50 microM in transport buffer. The validated methods were used to determine the transport of thalidomide by Caco-2 monolayers. The transport across the monolayers from the apical (A) to basolateral (B) side was similar to that from B to A side. The apparent permeability coefficient (P(app)) values of thalidomide at 10-300 microM from the A to B and from B to A side was 2-6 x 10( 5) cm/s, with a marked decrease in P(app) values from A to B side at increased thalidomide concentration. The A to B transport appears to be dependent on temperature and sodium ion. Sodium azide, 2,4-dinitrophenol (both ATP inhibitors), 5-fluorouracil, cytidine and glutamic acid significantly inhibited the transport of thalidomide. These results indicate that the transport of thalidomide by Caco-2 monolayers was rapid, which might involve an energy dependent mechanism. PMID- 12535850 TI - Validation of liquid chromatography assay for the quantitation of (Z)-3-[2,4 dimethyl-5-(2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-1H-pyrrol-3-yl]propionic acid (SU006668) in human plasma and its application to a phase I clinical trial. AB - The validation of an analytical method to quantify the antiangiogenic, (Z)-3-[2,4 dimethyl-5-(2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-1H-pyrrol-3-yl]propionic acid (SU006668) for pharmacokinetic determination in a phase I clinical trial, is described. HPLC, with a gradient mobile phase and UV detection at 440 nm, was used. SU006668 was extracted from plasma by precipitation of proteins with acetonitrile. The assay was linear from 25 to 2000 ng/ml (r(2)=0.997); sensitive (limit of quantification 25 ng/ml), accurate (RE 2.6-11.9%) and reproducible (inter-batch precision C.V. 3.2%). Pharmacokinetic data for six patients are presented. They show linear pharmacokinetics with a low volume of distribution and induction at doses of 50, 100 and 200 mg/m(2). PMID- 12535851 TI - Simultaneous determination of oleuropein and hydroxytyrosol in rat plasma using liquid chromatography with fluorescence detection. AB - Oleuropein, the main glycoside present in olives, and hydroxytyrosol, the principal degradation product of oleuropein present in olive oil, have been linked to reduction of coronary heart disease and certain cancers. In the present study a direct and sensitive reversed-phase high-performance liquid chromatographic assay was developed for simultaneous quantification of both oleuropein and hydroxytyrosol. The plasma protein was precipitated with acetonitrile, samples were then centrifuged and supernatants were dried, and reconstituted with water prior to injection. The chromatographic analysis was carried out using a phenyl column and an isocratic elution of acidified water and acetonitrile with fluorescence detection at 281 and 316 nm for excitation and emission, respectively. The calibration curve was linear and limits of quantification were 30 ng/ml and 3 microg/ml for hydroxytyrosol and oleuropein, respectively. The method has been successfully applied to monitor oleuropein and hydroxytyrosol plasma levels in the rat. PMID- 12535852 TI - Confirmation of ofloxacin precipitation in corneal deposits by microbore liquid chromatography-quadrupole time-of-flight tandem mass spectrometry. AB - We investigated the corneal precipitate of a 6-year-old boy with vernal keratoconjunctivitis (VKC), treated with topical ofloxacin 0.3% eyedrops. Because of the extremely small sample amount (corneal scraping), a very sensitive and specific method was needed with the possibility of an unambiguous identification of ofloxacin, supposed to be present in the precipitate. In this respect, tandem Q-TOF mass spectrometry combined with micro LC (1 mm I.D.) was chosen. Confirmation of the presence of ofloxacin in the deposit was obtained by means of the characteristic product ion spectrum produced by CID. This clearly indicated that the precipitate, removed by corneal scraping from the 6-year-old boy with VKC, contained ofloxacin. PMID- 12535853 TI - Different inhibitors of plasmin differentially affect angiostatin production and angiogenesis. AB - Plasmin is a broad-spectrum serine proteinase, which is presumed to cleave many extracellular proteins and affect angiogenesis. In the present work, we studied the effect of two different inhibitors of plasmin (epsilon-aminocaproic and alpha(2)-antiplasmin) on angiogenesis in vivo using the chicken embryo chorioallantoic membrane assay, and in vitro using human umbilical vein endothelial cells. Epsilon-aminocaproic acid inhibited, while alpha(2) antiplasmin induced, angiogenesis, as well as human umbilical vein endothelial cell proliferation, migration and tube formation on matrigel in a dose-dependent manner. Since plasmin has been implicated in the production of angiostatin, we studied the effect of the two plasmin inhibitors on angiostatin protein amounts in the chicken embryo chorioallantoic membrane. In this tissue, the 38- and 45 kDa isoforms of angiostatin are differentially affected by the two inhibitors: epsilon-aminocaproic acid increased, while alpha(2)-antiplasmin decreased the amounts of both isoforms. These data suggest that plasmin may have an antiangiogenic role in vivo through generation of angiostatin. Moreover, plasmin inhibitors differentially affect in vivo angiogenesis, depending on the mechanism by which they inhibit plasmin activity. PMID- 12535854 TI - Amphetamine enhances Ca2+ entry and catecholamine release via nicotinic receptor activation in bovine adrenal chromaffin cells. AB - Amphetamine, a psychostimulant, has been shown to act as a channel blocker of muscle nicotinic receptors and to induce a Ca(2+)-dependent secretion from adrenal chromaffin cells. In this study, the relationship between amphetamine and nicotinic receptors was studied using bovine adrenal chromaffin cells as a model system. Our results show that D-amphetamine sulfate alone induced an increase in the cytosolic Ca(2+) concentration ([Ca(2+)](c)) and [3H]norepinephrine release in a dose-dependent and extracellular Ca(2+)-dependent manner. Two common nicotinic receptor antagonists, hexamethonium and mecamylamine, suppressed the D amphetamine sulfate-induced [Ca(2+)](c) rise and [3H]norepinephrine release. In addition, D-amphetamine sulfate inhibited the 1,1-dimethyl-4-phenyl-piperazinium iodide (DMPP)-induced [Ca(2+)](c) rise and [3H]norepinephrine release, but not the high K(+)- or veratridine-induced [Ca(2+)](c) increase and [3H]norepinephrine release. Antagonists, including alpha-bungarotoxin and choline, that are more specific for alpha7 nicotinic receptors were capable of inhibiting the D amphetamine sulfate-induced [Ca(2+)](c) rise, while D-amphetamine sulfate was found to be capable of inhibiting the [Ca(2+)](c) rise induced by the alpha7 nicotinic receptor agonists, epibatidine and choline. Moreover, D-amphetamine sulfate dose-dependently suppressed [3H]nicotine binding to chromaffin cells. We, therefore, conclude that D-amphetamine sulfate acts as a nicotinic receptor agonist to induce [Ca(2+)](c) increase and [3H]norepinephrine release in bovine adrenal chromaffin cells. PMID- 12535855 TI - Beta 1-adrenoceptor selectivity of nebivolol and bisoprolol. A comparison of [3H]CGP 12.177 and [125I]iodocyanopindolol binding studies. AB - There is an ongoing discussion on whether or not high beta(1)-adrenoceptor selectivity of beta-adrenoceptor antagonists may be favorable in the treatment of patients with heart failure. The present study compared the beta(1)-adrenoceptor selectivity of nebivolol and bisoprolol with that of carvedilol in the human myocardium, using a binding assay in conjunction with either the hydrophilic ligand (+/-)-[3H]4-(3-tertiarybutylamino-2-hydroxypropoxy)-benzimidazole-2-on HCl ([3H]CGP 12.177) or the lipophilic ligand [125I]iodocyanopindolol as radiolabeled compound. Measurements were made using membrane preparations obtained from identical nonfailing donor hearts. beta-adrenoceptor density was found to be slightly higher when [125I]iodocyanopindolol was used compared to [3H]CGP 12.177 (256+/-15 and 213+/-18 fmol/mg protein, respectively). When the highly beta(1) adrenoceptor-selective compound 2-hydroxy-5-(2-(hydroxy-3-(4((1-methyl-4 trifluoromethyl)-1-H-imidazol-2-yl)-phenoxy)-propyl)-aminoethoxyl)-benzamide (CGP 20.712A) and the highly beta(2)-adrenoceptor-selective compound erythro-(+/-)-1 (7-methylindan-4-yloyl)-3-isopropylaminobutan-2-ol HCl (ICI 118.551) were used in competition experiments, a similar proportion of beta(1)-adrenoceptors was seen for [3H]CGP 12.177 (69.3+/-1.6%) and for [125I]iodocyanopindolol (67.0+/-2.1%). K(i)(beta(1)) and K(i)(beta(2)) were obtained in the presence of 50 nM ICI 118.551 and 300 nM CGP 20.712A. The rank order of beta(1)-adrenoceptor selectivity (K(i)(beta(2))/K(i)(beta(1)) ratio) was nebivolol (for [3H]CGP 12.177 46.1 and for [125I]iodocyanopindolol 22.5)>bisoprolol (13.1 and 6.4)>carvedilol (0.65 and 0.41). To investigate whether in vivo metabolized nebivolol retains high beta(1)-adrenoceptor selectivity, serum specimens were collected before and 2 h after oral administration of 5 mg nebivolol. The samples were used for [125I]iodocyanopindolol binding studies with the myocardial membrane preparations. In these samples, the binding of [125I]iodocyanopindolol to beta(1) adrenoceptors was inhibited by 46.4+/-5.3%, whereas the binding to beta(2) adrenoceptors was inhibited by 20.5+/-1.1% compared to that of control samples. It is concluded that nebivolol is approximately 3.5 times more beta(1) adrenoceptor-selective than bisoprolol in the human myocardium. Furthermore, in vivo metabolized nebivolol retains beta(1)-adrenoceptor selectivity. PMID- 12535856 TI - Alfentanil potentiates anaesthetic and electroencephalographic responses to ketamine in the rat. AB - The interactions between mu-opioid and N-methyl-D-aspartate (NMDA) receptors have important implications for clinical pain management. We recently examined the pharmacokinetics of ketamine in rats following i.v. infusion of ketamine (racemate, 50 mg/kg/5 min) and found increased central nervous system distribution of ketamine in the presence of low constant plasma alfentanil concentrations (approximately 50 ng/ml). We now report on the effects of low plasma alfentanil concentrations on the duration of anaesthetic and electroencephalographic (EEG) responses to i.v. infusion of ketamine. Compared to ketamine alone, alfentanil significantly increased both the duration of anaesthesia (by 130%, P=0.00022) and the processed EEG power (microV(2)/s) (by 48%, P=0.0040). The plasma ketamine concentration producing half-maximal EEG effect was significantly reduced (by 60%, P<0.0001) in the presence of alfentanil. The results indicate that low plasma alfentanil concentrations potentiate the anaesthetic and EEG effects produced by ketamine. PMID- 12535857 TI - (-)-Linalool produces antinociception in two experimental models of pain. AB - Linalool is a monoterpene compound commonly found as a major component of the essential oils of several aromatic plant species, many of which are used in traditional medical systems as analgesic and anti-inflammatory remedies. We previously reported that (-)-linalool, the natural occurring enantiomer, plays a major role in the anti-inflammatory activity displayed by different essential oils, suggesting that linalool-producing species are potentially anti inflammatory agents. In this study, the antinociceptive activity of (-)-linalool was examined in two different pain models in mice: the acetic acid-induced writhing response, a model of inflammatory pain, and the hot plate test, a model of supraspinal analgesia. Moreover, the effect of (-)-linalool on spontaneous locomotor activity (25, 50, 75 and 100 mg/kg) was evaluated. The results show that this compound induced a significant reduction of the acid-induced writhing at doses ranging from 25 to 75 mg/kg. Such effect was completely reversed both by the opioid receptor antagonist naloxone and by the unselective muscarinic receptor antagonist atropine. In the hot plate test, only the dose of 100 mg/kg of (-)-linalool resulted in a significant effect. (-)-Linalool induced a dose dependent increase of motility effects, thus ruling out the confounding influence of a possible sedative effect. The more pronounced effect of (-)-linalool on the writhing test with respect to the hot plate test is consistent with the observation that (-)-linalool possesses anti-inflammatory activity. Finally, the activation of opioidergic and cholinergic systems appears to play a crucial role in (-)-linalool-induced antinociception. PMID- 12535858 TI - Does Cl-/HCO3- exchange play an important role in reperfusion arrhythmias in rats? AB - The protective effects of Cl(-)/HCO(3)(-) exchange inhibitors, 4,4' diisothiocyano-stilbene-2,2'-disulfonic acid (DIDS) and 4-acetamido 4'isothiocyanato-stilbene-2,2'-disulfonic acid (SITS), against reperfusion induced arrhythmias were investigated in anesthetized rats. Rats were subjected to 5-min occlusion of the left coronary artery followed by 10-min reperfusion. All drugs were intravenously administered 5 min before the onset of occlusion. DIDS (75 mg/kg) reduced the incidence of ventricular fibrillation and mortality to 0%, whereas SITS (75 mg/kg) only decreased these parameters to 60%. DIDS simultaneously decreased the mean blood pressure and heart rate, and prolonged PQ and QRS intervals, whereas SITS produced a weaker effect on these parameters and no change in QRS interval. Mexiletine (5 mg/kg), which had been demonstrated to suppress the arrhythmias and reduce the heart rate and mean blood pressure in this model, was shown to prolong PQ and QRS intervals. Verapamil (0.5 mg/kg) or diltiazem (0.4 mg/kg) suppressed the arrhythmias, simultaneously decreasing the heart rate and mean blood pressure and prolonging PQ interval. The results indicate that the protective effect of DIDS on reperfusion arrhythmias in the anesthetized rats is unlikely to be attributed to the inhibitory action on Cl( )/HCO(3)(-) exchange, but possibly mediated by its blocking effects on cardiac ion channels, such as Na(+) or Ca(2+) channels. PMID- 12535859 TI - Cardiovascular effects of Y-27632, a selective Rho-associated kinase inhibitor, assessed in the halothane-anesthetized canine model. AB - Y-27632, (+)-(R)-trans-4-(1-aminoethyl)-N-(4-pyridyl) cyclohexanecarboxamide dihydrochloride monohydrate, is a selective Rho-associated kinase inhibitor, which has been suggested to possess multiple clinical applications based on the in vitro observations. Since information regarding in vivo cardiovascular effects of Y-27632 is still limited, we assessed them using the halothane-anesthetized, closed-chest canine model. Administration of Y-27632 in a dose of 0.01 mg/kg, i.v. significantly decreased total peripheral vascular resistance together with an increase of cardiac output without affecting other cardiovascular parameters. Moreover, additional administration of Y-27632 in a dose of 0.1 mg/kg, i.v. significantly decreased blood pressure and left ventricular end-diastolic pressure, increased the heart rate and cardiac contractility, enhanced atrioventricular conduction and shortened the repolarization process as well as the effective refractory period. These results indicate that Y-27632 exerts a potent arterio-venodilator action with cardiostimulatory effects possibly through the sympathetic reflex in the in vivo canine model. PMID- 12535860 TI - Anti-thrombin action of low-dose acetylsalicylic acid. AB - It is known that low-dose aspirin is effective in coronary artery therapy, although it has not yet been clarified how it exerts its action. Here, we report that treatment of coronary artery patients with 100 mg/day of aspirin does not attenuate thrombin generation, but reduces free thrombin by favouring the formation of thrombin/antithrombin (TAT) complexes. Antithrombin hyperactivation is mediated by inhibition of platelet factor 4 release from alpha-granules, leading to higher heparin availability. PMID- 12535861 TI - Elevation of histidine decarboxylase activity in the stomach of mice by ulcerogenic drugs. AB - Histamine is involved in the development of gastric lesions. To examine the contribution of the histamine-forming enzyme, histidine decarboxylase, to drug induced gastric lesions, we compared the effects of aspirin, indomethacin and dexamethasone on histidine decarboxylase activity in mice. Administration of these drugs, orally or intraperitoneally, elevated histidine decarboxylase activity in the stomach but not in the liver, lung or spleen, dexamethasone being the most potent. In contrast, acetaminophen (a non-ulcerogenic drug) was inactive. These results and our previously reported findings (elevation of histidine decarboxylase activity by lipopolysaccharide, interleukin-1 and tumour necrosis factor, and by different types of stress) suggest that an elevation of histidine decarboxylase activity in the stomach may be a common feature of the responses to ulcerogenic stimuli. The possible participation of histidine decarboxylase in gastric lesions is discussed on the basis of the known actions of histamine, our findings and the effect of histamine H(2) receptor antagonists on histidine decarboxylase activity. PMID- 12535862 TI - Dopamine D1 receptor agonists induce penile erections in rats. AB - The dopamine receptor agonist apomorphine has been recently introduced in the treatment of erectile dysfunction. While it is well established that dopamine D2 like receptors play a crucial role in this effect, conflicting result are reported in the literature as for the role of dopamine D1-like receptors. The aim of this study was to determine the effect of systemic administration of dopamine D1-like receptor agonists on penile erection in rats. Male Wistar rats were treated with three different, and not structurally related, dopamine D1-like receptor agonists: the partial agonists SKF38393 ((+) 2,3,4,5-tetrahydro-7,8 dihydroxy-1-phenyl-1H-3-benzazepine) and CY 208-243 ((-)-4,6,6a,7,8,12b-exahydro 7-methylindole [4,3-ab]fenantridine), and the full agonist A 77636 ((-)-(1R,3S)-3 Adamantyl-1-(aminomethyl)-3,4-dihydro-5,6-dihydroxy-1H-2-benzopyran hydrochloride). All three compounds dose-dependently increased the number of penile erections, with the full agonist A77636 showing a more pronounced effect with respect to the other two. Moreover, the dopamine D1-like receptor antagonist SCH 23390 ((R)-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3 benzazepine) dose-dependently antagonised A77636 effect. These results show that systemic administration of dopamine D1-like receptor agonists induce penile erection in rats. This observation suggests that dopamine D1-like receptor agonists might be considered as a possible alternative to apomorphine in the treatment of erectile dysfunction, thus avoiding the typical side effects related to the stimulation of dopamine D2-like receptors such as nausea. PMID- 12535863 TI - Differential modulation of murine lung inflammation by bradykinin B1 and B2 selective receptor antagonists. AB - The effect of bradykinin receptor antagonists was studied in a mouse (C57Bl/6) model of allergic lung inflammation. Bradykinin B(2) receptor antagonist HOE-140 (D-Arg-[Hyp(3),Thi(5),Dtic(7)-Oic(8)]bradykinin) or bradykinin B(1) receptor antagonist R-954 (Ac-Orn[Oic(2),alphaMePhe(5),betaD-Nal(7)Ile(8)]des-Arg(9) bradykinin) were given i.p. to ovalbumin sensitized mice 30 min before antigen challenge. After 24 h, bronchoalveolar lavage was performed for cell analysis and the lungs were removed for evaluation of airway hyperreactivity and histopathology. Treatment with HOE-140 caused a significant increase in bronchoalveolar lavage cell number: eosinophils (182%), neutrophils (98%), lymphocytes CD(4)(+) (192%), CD(8)(+) (236%), B220 (840%), Tgammadelta(+) (194%) and NK1.1(+) (246%). Hyperreactivity and mucus secretion were not significantly affected in this group. Treatment with R-954 significantly reduced eosinophil (79%) and neutrophil (83%) but has no effect on lymphocytes number in bronchoalveolar lavage fluid. Airway hyperreactivity and mucus secretion were reduced by this treatment (84% and 35%, respectively). These results show important modulatory effect of bradykinin B(1) and B(2) receptors on allergic lung inflammation. PMID- 12535865 TI - Nitric oxide synthase inhibition by N(G)-nitro-L-arginine methyl ester retards vascular sprouting in angiogenesis. AB - The present study was designed to evaluate the role of nitric oxide (NO) for angiogenesis. Angiogenesis was elicited upon mouse cornea by chemical cautery with silver nitrate. Angiogenic activity was evaluated by measuring the length of vascular sprout with or without administration of NO synthase (NOS) inhibitor, N((G))-nitro-L-arginine-methyl ester (L-NAME). In the pericorneal plexus, a circulatory loop situated in the same topological situation for all individuals was selected to observe vascular sprouting. At 72 h after cauterization, the length of the longest vascular sprout was measured using the perfused whole-mount cornea. The length of nontreated mice (83 +/- 83 microm) was significantly longer than that of L-NAME treated mice (33 +/- 24.6 microm). To address the possible contribution of production of vascular endothelial growth factor (VEGF) and NO, we measured mRNAs of VEGF and inducible NOS. The mRNA level of VEGF increased to 170% of the nontreated level at 12 h after cauterization and returned to the nontreated level by 24 h after cauterization. mRNA of inducible NOS remained elevated 24 h after cauterization. These results suggest that the response of preexisting vessels to angiogenic stimulus via NO is of importance in the process of angiogenesis, i.e., vascular sprouting is promoted by NO production. This might be attributable to enhancement of an increase in vascular permeability and /or vasodilation via NO. PMID- 12535866 TI - Inhibition of TIMP1 enhances angiogenesis in vivo and cell migration in vitro. AB - Neovascular invasion into a 3-dimensional matrix is controlled, in part, by matrix metalloproteinases (MMPs) and their inhibitors, tissue inhibitors of metalloproteinases (TIMPs). We tested the hypothesis that increasing MMP activity, via a specific blocking antibody to TIMP1, would enhance fibrovascular invasion into a PVA sponge. In vivo, inhibition of TIMP1 doubled the amount of angiogenic invasion (percentage area of invasion 33.5 +/- 3.5 vs 16.9 +/- 9.5, P = 0.003). The blocking antibody to TIMP1 did not increase the proportion of cells that were proliferating in the sponge implants, underscoring the importance of migration. In vitro, human microvascular endothelial cells (hmEC) and dermal fibroblasts treated with the antibody did not secrete greater amounts of collagenase but migrated significantly farther on collagen I (increase in distance migrated 26.6 +/- 9.4%, P = 0.003). Human dermal microvascular endothelial cells exposed to the TIMP1 blocking antibody exhibited a significant change in cell shape to a more elongated morphology. In conclusion, inhibition of TIMP1 increased angiogenesis into a PVA sponge in vivo and enhanced the migration of dermal hmEC and fibroblasts on collagen I in vitro. We propose that blocking TIMP1 improves angiogenesis by increasing cell motility during fibrovascular invasion. PMID- 12535867 TI - Retinal endothelial cells are more susceptible to oxidative stress and increased permeability than brain-derived endothelial cells. AB - Increased vascular permeability and oxidative stress are important in diabetic retinopathy. Because the cerebral microcirculation is much less affected in diabetes, our objectives are to compare: (1) glutathione peroxidase activity, (2) superoxide dismutase levels, (3) superoxide production, and (4) junctional protein (ZO-1) levels between retinal and brain-derived endothelial cells. Bovine brain and retinal endothelial cell cultures are incubated in medium containing either mM or 30 mM glucose for 5 days. Superoxide is measured in the medium and endothelial cells are then lysed and analyzed for glutathione peroxidase activity as well as levels of superoxide dismustase and ZO-1. The results demonstrate that, compared to brain-derived endothelial cells, retinal endothelial cells release high levels of superoxide, have less glutathione peroxidase activity and lower levels of superoxide dismutase, and ZO-1. Also, unlike brain-derived endothelial cells where ZO-1 levels increased in response to glucose, in retinal endothelial cells, ZO-1 levels are unaffected by glucose. These findings suggest that greater oxidative stress and lower junctional protein levels in retinal endothelial cells may contribute to blood/retinal barrier dysfunction in diabetic retinopathy. PMID- 12535868 TI - Pharmacological investigations on lipopolysaccharide-induced permeability changes in the blood-brain barrier in vitro. AB - Lipopolysaccharide-induced changes in blood-brain barrier (BBB) permeability were investigated with a pharmacological approach in vitro. Lipopolysaccharide induced a concentration- and time-dependent (non)reversible opening of the BBB, and brain astrocytes make brain capillary endothelial cells (BCEC) resistant to this BBB disruption. De novo protein synthesis was essential for the recovery, because cycloheximide prevented the recovery process. Dexamethasone pretreated BCEC were more resistant to lipopolysaccharide, while no protective response was induced by heat shock nor by inhibition of P-glycoprotein. BBB opening was tempered by free radical inhibitors (i.e., pretreatment with N-acetyl-cysteine or uric acid combined with deferroxamine mesylate). No effects of modulators of prostanoid-, leukotriene-, or platelet-activating factor pathways were observed. Therefore, lipopolysaccharide-induced BBB opening seems to be primarily mediated by excessive free radical production. PMID- 12535869 TI - pO(2) measurements by phosphorescence quenching: characteristics and applications of an automated system. AB - An automated system for pO(2) analysis based upon phosphorescence quenching was tested. The system was calibrated in vitro with capillary samples of saline and blood. Results were compared to a conventional measuring procedure wherein pO(2) was calculated off-line by computer fitting of phosphorescence decay signals. PO(2) measurements obtained by the automated system were correlated (r(2) = 0.98) with readings simultaneously generated by a blood gas analyzer, accuracy being highest in the low (0-20 mm Hg) and medium pO(2) ranges (21-70 mm Hg). Measurements in in vivo studies in the hamster skin-fold preparation were similar to previously reported results. The automated system fits the phosphorescence decay data to a single exponential and allows repeated pO(2) measurements in rapid sequence. PMID- 12535870 TI - Propranolol and atropine do not alter choroidal blood flow regulation during isometric exercise in healthy humans. AB - Recent studies indicate that the human choroid has a considerable capacity to keep blood flow constant despite exercise-induced increases in perfusion pressure. The mechanisms underlying this vasoconstrictor response remain unclear. We hypothesized that pharmacological modulation of the autonomic nervous system may alter the choroidal pressure/flow relationship during squatting. To test this hypothesis, we performed a randomized, double-masked, placebo-controlled, three way crossover study in 15 healthy male volunteers. Subjects received, on different study days, intravenous infusions of the beta-adrenoceptor antagonist propranolol, the muscarinic receptor antagonist atropine, or placebo. During these infusions, subjects performed squatting for 6 min. Choroidal blood flow was assessed with laser Doppler flowmetry and ocular perfusion pressure (OPP) was calculated from mean arterial pressure and intraocular pressure. As expected, propranolol reduced basal pulse rate, whereas atropine increased pulse rate, indicating that the drugs were administered at systemically effective doses. None of the drugs altered the choroidal pressure/flow relationship during isometric exercise. These data indicate that the regulatory vasoconstrictor capacity of the choroid during exercise is not affected by systemic blockade of beta adrenoceptors or muscarinic receptors. PMID- 12535871 TI - Angiogenic stimulus determines the positioning of pericytes within capillary sprouts in vivo. PMID- 12535872 TI - Blood flow analysis in mesenteric microvascular network by image velocimetry and axial tomography. PMID- 12535873 TI - A model for temporal heterogeneities of tumor blood flow. AB - Tumor blood flow (TBF) plays a fundamental role in tumor growth and treatment, and is characterized by spatial and temporal heterogeneities. Here we show that the interstitial fluid pressure (IFP), which is higher in tumoral tissue than in normal tissue, coupled with the tumor microvascular pressure (MVP) and the higher permeability of tumoral vessels, can explain the sustained oscillatory behavior of TBF, observed in vivo. PMID- 12535874 TI - Mediastinal fat: a source of cells for tissue engineering of coronary artery bypass grafts. PMID- 12535875 TI - Laser-Doppler imaging of osteoarthritis in proximal interphalangeal joints. PMID- 12535900 TI - Proceedings of the Symposium on Graduated Diver Licensing: Documenting the Science of GDL. Chatham, Massachusetts, USA. November 5-7, 2002. PMID- 12535901 TI - Teenage drivers: patterns of risk. AB - PROBLEM: To determine patterns of risk among teenage drivers. METHOD: Review and synthesis of the literature. RESULTS: On most measures, crash rates during the teenage years are higher than at any other age, for both males and females. Risk among teenagers varies greatly by driving situation; it is particularly low in some situations (e.g., the learner period) and particularly high in others (e.g., right after licensure, late at night, with passengers present). In some of these high-risk driving situations, risk is elevated for drivers of all ages (e.g., late night driving), in others risk is elevated more for teens than adults (e.g., driving after consuming alcohol), and in others the risk is unique to teen drivers (e.g., having passengers). IMPACT ON RESEARCH, PRACTICE, AND POLICY: These varying patterns of risk form the basis for graduated licensing systems, which are designed to promote low-risk and discourage high-risk driving. PMID- 12535902 TI - The genesis of GDL. AB - This paper discusses the early research that lead to graduated driver licensing, some of the educational principals on which it is based, obstacles to its acceptance, and some of the early efforts in the U.S. and elsewhere. EARLY RESEARCH: The research underlying the concept of graduated driver licensing was a 1971 North Carolina study that identified the overrepresentation of young drivers in crashes at night and when another young person was the right front passenger. EDUCATIONAL PRINCIPALS: Efforts to reduce the risk to young novice drivers applied what was known about learning. The concepts included distributed learning (i.e., over time) and progressing from simple to complex skills. A PROPOSAL: The proposed graduated licensing system based on learning principals included (a) initial experience under low risk conditions, (b) extended supervised practice, (c) gradual move to more complex conditions, and (d) harsher penalties for deliberate risk-taking. OBSTACLES: There were several most common objections raised against graduated licensing. Raising the licensing age decreased mobility. Some young drivers were "good" drivers. Enforcement is difficult. Fear of parental objections. Parents are not driver educators and some young people do not have an available parent. Administrative costs are too high. ACCEPTANCE: Driver educators were the first to see the benefits of a graduated system in the 1970s and 1980s. Toronto nearly adopted a graduated system in 1976. New Zealand was the first to adopt a graduated licensing system in 1984. Michigan in 1997 was the first state to require parental certification of extended supervised driving practice. PMID- 12535903 TI - The evolution and effectiveness of graduated licensing. AB - This paper traces the history of graduated licensing, starting about the point in time when Pat Waller's paper on the genesis of the concept ends, and examines the extent to which graduated licensing has produced reductions in collisions. It concludes with some general observations about future research needs, anticipating several of the papers that follow. The evolution of graduated licensing is chronicled, beginning with the early and largely unsuccessful efforts to introduce it in the United States in the late 1970s, through the pioneering efforts in New Zealand, which resulted in the first truly graduated system in 1987, to Canada where the program was introduced 7 years later, to the United States where it has flourished in more recent years. This 25-year history lesson hopefully creates an appreciation for the somewhat torturous journey that graduated licensing has experienced in achieving acceptance among the public and policy-makers-a journey that is not yet over, as subsequent papers in the symposium will show. The proliferation of graduated licensing in recent years is a mixed blessing-the wider adoption of graduated licensing has been a very positive development, but the programs that have evolved are anything but homogeneous in structure or content. Although this is often necessary for various reasons, it is worrisome that some programs are graduated licensing in name only. This suggests that future efforts to promote graduated licensing must emphasize adherence to the fundamental risk reduction and multistage principles on which the concept is based. The paper also considers the extent to which graduated licensing achieves its objective of reducing collisions among those covered by the program. Understandably, most jurisdictions would not introduce graduated licensing until it was shown to be effective and this, to some extent, slowed the process of implementation. The obvious irony is that it could not be shown to be effective until it was introduced. Fortunately, as history demonstrates, some jurisdictions were prepared to try the system based on its very sound empirical rationale. And, their confidence has been rewarded. A growing body of research, which shows that graduated licensing has been associated with significant and substantial reductions in collisions, is briefly described. The paper concludes with some general observations designed to anticipate the papers that follow. First, it outlines questions that still remain unanswered about graduated licensing-why or how it works, with whom it works, and what features are most effective. Precise and unambiguous answers to these questions are essential for the design of a system that maximizes the potential for reducing collisions, injuries, and deaths. Second, it signals a note of caution on the limits of graduated licensing-it is important to recognize just how effective and beneficial this program is; it is equally important to recognize that it is not the sole panacea for the problem of collisions involving new drivers. PMID- 12535904 TI - The learner's permit. AB - PROBLEM: This paper considers the role and value of an extended learner's period in a graduated licensing system. METHOD: Review and synthesis of the literature. RESULTS: The learner's permit allows beginners to practice under supervision before attempting the road test for a driver's license. A learner's permit stage was an integral part of the "provisional" (graduated) licensing model initially formulated by NHTSA in the mid-1970s. Almost all Canadian provinces and U.S. states now have graduated licensing (GDL) programs that typically include an extended period of supervised driving. Most parents and teens favor the learner stage, and consistent with the intent of GDL, most learners are practicing and gaining driving experience under low-risk conditions. Research shows that very few learners crash while under supervision and that an extended learner stage has safety benefits. IMPACT ON RESEARCH, PRACTICE AND POLICY: Further research is needed on the safety benefits of an extended learner stage and on its optimal features. PMID- 12535905 TI - Provisional license. AB - The Provisional or Intermediate license is the central phase in a comprehensive Graduated Licensing System. Young drivers qualify for a Provisional license by completing their Learners phase, reaching a specified age, and passing a road test. Provisional license holders are allowed to drive unsupervised subject to various restrictions. Typical restrictions include prohibitions against night driving, passengers and alcohol. The goal is to limit teen exposure to risky driving situations during their first few months of licensure, a time when their crash rates are extremely high. States with such restrictions have been shown to have lower teen crash rates than states without. Parents indicate strong support both for Graduated Licensing Systems and for the specific restrictions. PMID- 12535906 TI - The provisional license: nighttime and passenger restrictions--a literature review. AB - PROBLEM: Due to inexperience and inadequate driving skills, the road is a very risky place for young and beginning drivers, yet such experience and skills can only be built by increased driving and exposure to risks on the road. Graduated driver licensing (GDL) allows beginning drivers to get their initial driving experience under less risky conditions and gradually eases them into more complex driving situations. This paper reviews the literature exploring two key features of the intermediate licensing phase of GDL, nighttime driving restrictions and passenger restrictions. METHOD: Literature review. RESULTS: Nighttime driving restrictions have been shown to effectively reduce the number and rate of crash involvements on the part of teenage drivers. Data suggest that having passengers in the car increases the likelihood of a fatal injury in young drivers and that this risk increases with the number of passengers. Young drivers were more likely to cause a crash when accompanied by their peers. DISCUSSION: Nighttime driving and passenger restrictions are effective in decreasing injuries among teenage drivers and their passengers, especially in the context of a full GDL system. Several research questions remain to be answered in order to fully refine and optimize the impact of these provisional measures. PMID- 12535907 TI - Graduated driver licensing in the United States: evaluation results from the early programs. AB - BACKGROUND: Seventeen states enacted graduated driver licensing (GDL) programs that were implemented from 1996 through 1999 and for which evaluations are of interest. METHODS: We received evaluation results reported for six states for which data were available. Summarizing results is difficult in other than the most global terms because of differences in pre-GDL programs, differences in GDL programs, and differences in evaluation methodology. RESULTS: All states identified some crash reduction among teen drivers following GDL implementation. This positive effect was observed across different geographic regions, and with different GDL programs. Simple counts are down-fewer teens are experiencing crashes and becoming injured. After calculating crash rates to adjust for changes over time in populations or licensed drivers, reductions generally were still found. Population-adjusted risks of injury/fatal crash involvement of 16-year-old drivers in Florida and Michigan were reduced by 11% and 24%, respectively. Population-adjusted risks of any crash involvement of 16-year-old drivers in Michigan and North Carolina were reduced by 25% and 27%, respectively. Reductions in night (restricted hours) crash risk were impressive in Florida, Michigan, and North Carolina. A comparison state design was only possible in the Florida evaluation, and results showed greater crash reductions under GDL. Change-point analyses of Michigan's crash data trends over time provided additional support of GDL's effectiveness in reducing crashes. DISCUSSION: Taken as a whole, and including the preliminary findings from California, Ohio, and Pennsylvania, these reports demonstrate the early effectiveness of GDL in reducing the crash risk of teen drivers. The impact of these studies and others to come will guide future research, practice, and policy. PMID- 12535908 TI - Other high-risk factors for young drivers--how graduated licensing does, doesn't, or could address them. AB - PROBLEM: Young drivers, particularly those who are newly licensed, have a very high crash risk. This paper examines the risk factors underlying their high crash rates and assesses the extent to which existing graduated licensing programs address these risks and whether improvements to these programs should be considered. METHOD: Review and synthesis of the literature. RESULTS: The elevated risk among young drivers of being in an injury crash is the result of a number of factors found alone or in combination, such as risky driving, alcohol use, seat belt nonuse, driver distraction, fatigue, and vehicle choice. Nighttime and passenger restrictions, adopted widely in the United States, work by keeping drivers out of hazardous situations rather than by addressing risk factors directly. However, the risk factors remain in play in driving situations not specifically restricted by law. Although other graduated licensing components adopted around the world-more stringent exit tests (i.e., you need to pass a test to move to the next stage), hazard perception tests, and restrictions on speed, vehicle power, and roadway access-make sense based on the identified risk factors, they are not yet supported by research. Should research findings warrant it, consideration should be given to providing guidance to parents about how to keep their beginning drivers safe, including information on vehicle choice. IMPACT ON RESEARCH, PRACTICE, AND POLICY: Researchers should continue to monitor and to evaluate innovative approaches to reduce the crash risk of young drivers. The effectiveness of new approaches should be established before adoption on a wider scale takes place. PMID- 12535909 TI - Enhancing the effectiveness of graduated driver licensing legislation. AB - PROBLEM: Many states have enacted graduated driver licensing (GDL) systems in an effort to reduce the very high crash rates of young beginning drivers. This article addresses how to achieve the maximum benefit from GDL by ensuring compliance with protective restrictions. ENHANCING GDL THROUGH SYSTEM STRUCTURE: The major crash reductions due to GDL systems result from the protective restrictions during the initial two levels, which isolate novice drivers from the highest risk driving situations. Accordingly, GDL systems should include protective restrictions that adequately control the greatest dangers facing young drivers: multiple teen passengers and night driving before midnight. ENCOURAGING COMPLIANCE THROUGH SYSTEM STRUCTURE: Including protective restrictions that are supported by parents and teens will encourage compliance. Furthermore, linking a teen's advancement through GDL to demonstrated responsible driving will likely encourage compliance more than threatening punishment for violations. ENCOURAGING COMPLIANCE THROUGH ENFORCEMENT: Parents are in a prime position to enforce most GDL restrictions, but there is some evidence they do not enforce those restrictions that they consider too extreme. Little is currently known about the involvement of law enforcement in GDL systems, but there is potential for high visibility law enforcement activities to encourage compliance with restrictions. IMPACT ON RESEARCH, PRACTICE, AND POLICY: There is a need for better designed GDL systems in many states; more research is needed to examine compliance with restrictions and to evaluate enforcement efforts by parents and law enforcement. PMID- 12535910 TI - Graduated driver licensing and safer driving. AB - Graduated Driver Licensing (GDL) inserts between the leaner permit and full licensure an intermediate or "provisional" license that allows novices to drive unsupervised but subject to provisions intended to reduce the risks that accompany entry into highway traffic. Introduction of GDL has been followed by lowered accident rates, resulting from both limiting exposure of novices to unsafe situations and by helping them to deal with them more safely. Sources of safer driving include extended learning, early intervention, contingent advancement, and multistage instruction. To extend the learning process, most GDL systems lengthen the duration of the learner phase and require a specified level of adult-supervised driving. Results indicate that extended learning can reduce accidents substantially if well structured and highly controlled. Early intervention with novice traffic violators have shown both a general deterrent effect upon novice violators facing suspension and a specific effect upon those who have experienced it. Making advancement to full licensure contingent upon a violation-free record when driving on the provisional license has also evidenced a reduction in accidents and violations during that phase of licensure. Multistage instruction attempts development of advanced skills only after novices have had a chance to master more basic skills. Although this element of GDL has yet to be evaluated, research indicates crash reduction is possible in situations where it does not increase exposure to risk. While the various elements of GDL have demonstrated potential benefit in enhancing the safety of novice drivers, considerable improvement in the nature and enforcement of GDL requirements is needed to realize that potential. PMID- 12535911 TI - How well do parents manage young driver crash risks? AB - Motor vehicle crashes are extremely high among young drivers during at least the first year of licensure. Crash risks decline with increased experience, but the more newly licensed teenagers drive, the greater their risk exposure. Hence, the dilemma facing policy makers and parents is how to provide young drivers with driving experience without unduly increasing their crash risk. Graduated driver licensing policies serve to delay licensure and then limit exposure to the highest risk conditions after licensure, allowing young drivers to gain experience only under less risky driving conditions. A similar strategy is needed to guide parents. Parents do not appear to appreciate just how risky driving is for novice drivers and tend to exert less control over their teenage children's driving than might be expected. Recent research has demonstrated that simple motivational strategies can persuade parents to adopt driving agreements and impose greater restrictions on early teen driving. PMID- 12535912 TI - Graduated driver licensing: the New Zealand experience. AB - In New Zealand, on 1 August 1987, a three-stage graduated driver licensing (GDL) system that applied to all new drivers aged 15-24 years was introduced. The essential elements of GDL were a 6-month learner license (supervised driving) and an 18-month restricted license stage (with restrictions on night driving and carrying passengers). A blood alcohol limit of 0.03 mg% applied at both stages. EVALUATION STUDIES: Early studies indicated that young people were reasonably accepting of the restrictions, with the passenger restriction being the least acceptable. Problems of compliance with the restricted license driving restrictions were reported. Evaluations of the impact of the graduated driver licensing (GDL) on serious traffic-related injury showed that up until 1991-1992, an 8% reduction could be attributed to GDL. At this time, it was considered that reduced exposure was the main reason for this reduction. However, the number of fatalities and hospital admissions among young people continued to decline, as did the population rate and the rate per number of licensed drivers among the young driver age group. A further evaluation study showed that drivers with a restricted license had a smaller proportion of crashes at night, and with passengers, compared with drivers licensed before GDL. IMPACT OF GDL: These results suggested that GDL restrictions had contributed to the reduction in crashes among young people and that it was not simply a case of reduced exposure to risk. An update of the most recent crash statistics indicated that, compared with older age groups, the fatal and serious injury crash rate among young people has remained substantially below the pre-GDL level. This suggests that the impact of GDL has not diminished over time. PMID- 12535913 TI - What we know, what we don't know, and what we need to know about graduated driver licensing. AB - On November 5-7, 2002, the Symposium on Graduated Driver Licensing in Chatham, MA, brought together 75 researchers and practitioners from the United States, Canada, Australia, and New Zealand to document the current science of graduated driver licensing (GDL) and to outline research needs. Participants reviewed 12 background papers and discussed the papers in depth. The symposium's background papers are published in this issue of the Journal of Safety Research. This paper summarizes and provides a quick reference to information from the symposium papers and participant discussions. It cites the 12 symposium papers, which in turn provide more information and cite original sources. Issues and recommendations not followed by a citation were raised in the symposium discussions. This paper is divided into seven sections. The first six sections summarize information from the symposium papers and discussions. The sections are: (1) The need for graduated driver licensing; (2) Effectiveness of GDL as implemented; (3) The learner's permit phase; (4) The provisional license phase; (5) The roles of teens, parents, and public agencies; and (6) Enacting and implementing GDL. In each of these six sections, research needs are classified as either high priority (important for designing and implementing effective GDL programs) or lower priority (useful but not critical for GDL at this time). The final section summarizes the discussion of research issues and priorities from the symposium's closing session. This section has three topics: general research, issues involving parents, and issues involving graduated licensing legislation and implementation. It presents participants' collective views on both broad priorities and specific issues. In providing a concise summary of presentations and discussions from the symposium, this paper necessarily omits some information and points of discussion. The views and judgments expressed are the authors' best attempt to capture the symposium's consensus, but they do not necessarily represent the views of the authors, their organizations, or any other individual symposium participant. In particular, they are not necessarily endorsed by the symposium's sponsors: General Motors, the National Highway Traffic Safety Administration, the National Safety Council, and Nationwide. PMID- 12535914 TI - Speech by the Hon. Carol J. Carmody. PMID- 12535915 TI - Portraying Caroline Herschel. PMID- 12535917 TI - Endeavour's greatest Briton. PMID- 12535918 TI - Cyrus Levinthal, the Kluge and the origins of interactive molecular graphics. AB - In the mid-1960s, a group of scientists at Massachusetts Institute of Technology, led by Cyrus Levinthal, took hold of one of the early interactive graphics terminals and used it to visualize, study and model the structure of proteins and nucleic acids. From this encounter between cutting-edge computer technology and molecular biology emerged the crucial elements for the development of a research technology field known today as interactive molecular graphics. The following account is not only about how computer graphics technology has literally changed the way scientists view the molecular realm, but also a look at how an epistemic and institutional space was created to integrate this technology into scientific research. PMID- 12535919 TI - Mineralogy and mineral collections in 18th-century France. AB - During the 18th century, mineralogy constituted an integral part of natural history, sharing the concerns of botany and zoology over collection and classification. In Paris, many people owned private mineral collections, but these have been largely neglected by historians. Here, I examine the place of private collections in the history of mineralogy, arguing that they contributed socially, economically and intellectually to the field in a period before the dominance of the large national collection. I also show how the interests of private collectors diverged from those of the curators of public collections, particularly following the French Revolution. PMID- 12535920 TI - A brief history of x-rays. AB - The identity of x-rays was initially mysterious, their connotations including morbidity and the otherworldly. Gradually they became routine, finding a place in the high-energy end of the electromagnetic spectrum. Radiologists established a niche in hospitals, contributing to the industrialization of healthcare. Control of x-ray tube output was improved while an infrastructure enforcing radiation protection accompanied the continued growth of the field. X-ray diffraction proved a powerful tool with which to examine substances at the atomic scale. Since 1950, x-ray researchers have taken advantage of synchrotrons' copious production of continuous x-ray spectra, and the marriage of x-rays with electronics has led to a cornucopia of cheap and highly efficient analytical tools. PMID- 12535921 TI - Of frogs and men: the origins of psychophysiological time experiments, 1850-1865. AB - Towards the end of the 1840s, Hermann von Helmholtz began to investigate experimentally the propagation of stimuli within nerves. Helmholtz's experiments on animals and human subjects opened a research field that in the following decades was intensively explored by neurophysiologists and experimental psychologists. Helmholtz's pioneering investigations justify the central place he occupies in accounts of the history of modern psychophysiology. Studying the concrete experimental settings and their local contexts shows how deeply the work of scholars such as Helmholtz is embedded in the history of culture and technology. In particular, the rapidly growing technologies of electromagnetism, which gave rise to telegraphy and electric clocks, facilitated the time measurements of 19th-century physiologists and psychologists. PMID- 12535922 TI - Linus Pauling and the scientific debate over fallout hazards. AB - From 1954 to 1963, numerous scientists engaged in a public debate over the possible hazards from radioactive fallout from nuclear weapons testing. Nobel laureate Linus Pauling, a California Institute of Technology chemist, was one of the most prominent. His scientific papers relating to the fallout debate reveal many of the scientific, social and political issues involved in the controversy. Although the public controversy ended after the signing of the 1963 Limited Test Ban Treaty, many of the scientific questions about the possible hazards of low level radiation remain under debate within the scientific community. Moreover, the fallout debate was a prototype of current controversies over environmental and public-health hazards. PMID- 12535923 TI - Reform in the garden. AB - English gardens powered by steam engines offer an intriguing view of how technological and scientific progress was naturalized in the landscape of 19th century Prussia, and in Prussian culture, during the early years of industrial modernization. One such garden is Peacock Island, in the Havel River near Potsdam. A product of the reform era that preceded and followed the Napoleonic Wars, it expressed the goal of Friedrich Wilhelm III and his government to enter into the British system of factory production and world trade. Modern science, as represented especially by Alexander von Humboldt, played a prominent role. PMID- 12535930 TI - Connective tissue growth factor in Alzheimer's disease. PMID- 12535931 TI - Peripheral administration of lipopolysaccharide enhances the expression of guanosine triphosphate cyclohydrolase I mRNA in murine locus coeruleus. AB - GTP cyclohydrolase I is the first and rate-limiting enzyme for the de novo biosynthesis of tetrahydrobiopterin, which is the cofactor for tyrosine hydroxylase. Lipopolysaccharide can modulate tetrahydrobiopterin production by upregulating GTP cyclohydrolase I protein expression in the locus coeruleus in the mouse brain. The increased supply of tetrahydrobiopterin in the locus coeruleus leads to increased tyrosine hydroxylase activity without affecting the level of tyrosine hydroxylase protein expression, resulting in an increase in norepinephrine turnover at the site. This study was performed to address whether the increase in GTP cyclohydrolase I protein is dependent on the de novo synthesis of GCH in the locus coeruleus. After i.p. administration of lipopolysaccharide, the mRNA expression of GTP cyclohydrolase I was examined. The expression level increased within 2 h, and reached to maximum level at 4 h after the lipopolysaccharide administration. However, the mRNA expression level of 6 pyruvoyl-tetrahydropterin synthase and sepiapterin reductase, both of which are involved successively after GTP cyclohydrolase I in tetrahydrobiopterin biosynthesis, were not affected by the lipopolysaccharide administration. These results suggest that GTP cyclohydrolase I upregulation alone is enough to modulate tetrahydrobiopterin production in the locus coeruleus. In addition, the mRNA level of tyrosine hydroxylase was also not affected by the lipopolysaccharide administration. Taken together, the data indicate that GTP cyclohydrolase I plays a crucial role in regulating norepinephrine biosynthesis by a pathway the activity of which is triggered by lipopolysaccharide i.p. administration. PMID- 12535932 TI - Serotonin persistently activates the extracellular signal-related kinase in sensory neurons of Aplysia independently of cAMP or protein kinase C. AB - Activation of the extracellular signal-related kinase is important for long-term increases in synaptic strength in the Aplysia nervous system. However, there is little known about the mechanism for the activation of the kinase in this system. We examined the activation of Aplysia extracellular signal-related kinase using a phosphopeptide antibody specific to the sites required for activation of the kinase. We found that phorbol esters led to a prolonged activation of extracellular signal-related kinase in sensory cells of the Aplysia nervous system. Surprisingly, inhibitors of protein kinase C did not block this activation. Serotonin, the physiological transmitter involved in long-term synaptic facilitation, also led to prolonged activation of extracellular signal related kinase, but inhibitors of protein kinase A or protein kinase C did not block this activation. We examined whether the protein synthesis-dependent increase in excitability stimulated by phorbol esters was dependent on phorbol ester activation of extracellular signal-related kinase, but increases in excitability were still seen in the presence of inhibitors of extracellular signal-related kinase activation. Our results suggest that prolonged phosphorylation of extracellular signal-related kinase in the Aplysia system is not mediated by either of the classic second messenger activated kinases in this system, protein kinase A or protein kinase C and that extracellular signal related kinase is not important for phorbol ester induced long-term effects on excitability. PMID- 12535934 TI - Transient transfection protects PC6-3 cells from apoptosis induced by nerve growth factor deprivation. AB - Some mammalian neurons undergo apoptosis after neurotrophin deprivation. We studied neuronally differentiated PC6-3 pheochromocytoma cells, which are highly dependent on nerve growth factor for survival. We found that transient transfection with green fluorescent protein or beta-galactosidase protected cells from apoptosis induced by nerve growth factor deprivation. Individual transfection reagent components did not produce increased viability of nerve growth factor-deprived cells. This apparent neuroprotective effect from transient transfection was specific to neurotrophin deprivation, as cells treated with H(2)O(2) or staurosporine were not protected. To determine the mechanism of neuroprotection after transfection, the transfection status of identified groups of cells was assessed both before and after nerve growth factor deprivation. The results were consistent with a model whereby cells that are transfected but not yet expressing the transfected protein are relatively protected from nerve growth factor deprivation. We suggest that apoptosis induced by neurotrophin deprivation may interact with processes of transient transfection and expression of foreign genes in neuronal cells. Not only should these interactions be considered in transient transfection studies of neurotrophin-deprived neurons, but also their elucidation could lead to novel methods for achieving neuroprotection. PMID- 12535933 TI - Cocaine-induced proliferation of dendritic spines in nucleus accumbens is dependent on the activity of cyclin-dependent kinase-5. AB - Repeated exposure to cocaine produces an enduring increase in dendritic spine density in adult rat nucleus accumbens. It has been shown previously that chronic cocaine administration increases the expression of cyclin-dependent kinase-5 in this brain region and that this neuronal protein kinase regulates cocaine-induced locomotor activity. Moreover, cyclin-dependent kinase-5 has been implicated in neuronal function and synaptic plasticity. Therefore, we studied the involvement of this enzyme in cocaine's effect on dendritic spine density. Adult male rats, receiving intra-accumbens infusion of the cyclin-dependent kinase-5 inhibitor roscovitine or saline, were administered a 28-day cocaine treatment regimen. Animals were killed 24-48 h after the final cocaine injection and their brains removed and processed for Golgi-Cox impregnation. Our findings demonstrate that roscovitine attenuates cocaine-induced dendritic spine outgrowth in nucleus accumbens core and shell and such inhibition reduces spine density in nucleus accumbens shell of control animals. These data indicate that cyclin-dependent kinase-5 is involved in regulation of, as well as cocaine-induced changes in, dendritic spine density. PMID- 12535935 TI - Evidence of melanin-concentrating hormone-containing neurons supplying both cortical and neuroendocrine projections. AB - In the rat, melanin-concentrating hormone-containing projections are detected in the median eminence and in the neural lobe of the pituitary. After vascular injections of the retrograde tracers fluorogold or fastblue, melanin concentrating hormone neurons are retrogradely labeled in the rostromedial zona incerta and adjacent perifornical region. These neurons may be the source of the melanin-concentrating hormone projections toward the median eminence and posterior pituitary, and may release their secretory products into the bloodstream. After fastblue injections in the cerebral cortex and vascular fluorogold injections, some melaninconcentrating hormone neurons contain both tracers, indicating that they send collaterals in the cerebral cortex and in the median eminence/posterior pituitary. No such collaterals have been described for the classical neuroendocrine systems. The melanin-concentrating hormone system is thought to play a role in arousal in correlation with specific goal oriented behaviors such as feeding or reproduction. Some MCH neurons may be involved in such functions by modulating directly cortical activity as well as being neuroendocrine. PMID- 12535936 TI - Generalized epilepsy with febrile seizures plus type 2 mutation W1204R alters voltage-dependent gating of Na(v)1.1 sodium channels. AB - Nine mutations that cause generalized epilepsy with febrile seizures plus have been identified in the SCN1A gene encoding the alpha subunit of the Na(v)1.1 voltage-gated sodium channel. The functional properties of two of these mutations (T875M and R1648H) have previously been described. T875M was shown to enhance slow inactivation, while R1648H dramatically accelerated recovery from inactivation. In this report, we have cloned, expressed and characterized the functional effects of a third generalized epilepsy with febrile seizures plus mutation, W1204R (Am J Hum Genet 68 (2001) 866). The mutation was cloned into the orthologous rat channel, rNa(v)1.1, and at the same time a single base pair insertion at base 120 in the original rNa(v)1.1 clone was corrected. The level of expression of the corrected wild-type rNa(v)1.1 was approximately 1000-fold higher than that of the original clone and comparable to that achieved with other neuronal sodium channels expressed in Xenopus oocytes. The properties of the W1204R mutant in the corrected rNa(v)1.1 were determined in the absence and presence of the beta1 subunit in Xenopus oocytes. The W1204R mutation resulted in approximately 11 mV hyperpolarized shifts in the voltage-dependence of activation and steady-state inactivation when expressed as an alpha subunit alone. When the channels were coexpressed with the beta1 subunit, the hyperpolarized shifts were still present but smaller, approximately 5 mV in magnitude. All other properties that we examined were comparable for the mutant and wild-type channels. The negative shift in activation would increase channel excitability, whereas the negative shift in inactivation would decrease excitability. The negative shifts in both properties also shifted the window current, which is the voltage region in which sodium channels can continue to open because some percentage of channels are activated and not all of the channels are inactivated. The shift in window current for the W1204R mutation could result in hyperexcitability because the neuron's potential is more likely to reach the more negative range. These results demonstrate that a third SCN1A mutation that causes generalized epilepsy with febrile seizures plus 2 alters the properties of the sodium channel in a different manner than the previous two mutations that were studied. The diversity in functional effects for these three mutations indicates that a similar clinical phenotype can result from very different underlying sodium channel abnormalities. PMID- 12535937 TI - Administration of brain-derived neurotrophic factor suppresses the expression of heat shock protein 27 in rat retinal ganglion cells following axotomy. AB - Optic nerve transection results in the apoptotic cell death of the majority of retinal ganglion cells by 14 days. The neurotrophin brain-derived neurotrophic factor (BDNF) enhances survival of retinal ganglion cells. In addition, the small heat shock protein Hsp27, with its anti-apoptotic effects, may be important for neuron survival following axotomy or trophic factor withdrawal. We recently reported the induction and expression of Hsp27 in a subset of retinal ganglion cells following axotomy. Here we have examined the effect of BDNF administration on the expression of Hsp27 in axotomized adult rodent retinal ganglion cells. Retinal ganglion cells were pre-labeled with Fluorogold prior to optic nerve transection and concomitant intraocular injection of BDNF or vehicle. Hsp27 immunofluorescence was examined in retinal sections from 4 to 28 days following injury. Consistent with previous survival studies, the number of Fluorogold labeled retinal ganglion cells declined from 100% at 4 days to approximately 15% by 14 days following axotomy and vehicle injection. In contrast, with BDNF administration, retinal ganglion cell survival was maintained at 100% to 7 days following axotomy. We report that the number of Hsp27-positive injured retinal ganglion cells, as detected by immunohistochemical staining, was decreased by 50% in BDNF-treated retinas, when compared with vehicle-treated controls. This decreased expression of Hsp27 in response to BDNF treatment was seen both at early (4 days) and delayed (14 days) times. BDNF following optic nerve transection significantly reduced the expression of Hsp27 in retinal ganglion cells. These results indicate that BDNF may down-regulate alternate cell survival pathways, including the stress-induced expression of Hsp27, and may help to explain the failure of chronic neurotrophin treatment to maintain long-term retinal ganglion cell survival. PMID- 12535938 TI - Peroxynitrite generated in the rat spinal cord induces apoptotic cell death and activates caspase-3. AB - We previously demonstrated that the peroxynitrite concentration increases after impact spinal cord injury. This study tests whether spinal cord injury-elevated peroxynitrite induces apoptotic cell death. Peroxynitrite was generated at the concentration and duration produced by spinal cord injury by administering S morpholinosydnonimine through a microdialysis fiber into the gray matter of the rat spinal cord. Fragmented DNA was visualized by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling. Transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling-positive neurons were quantitated by counting the transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling and neuron-specific enolase double-stained neurons along the fiber track in the sections removed at 6, 12, 24 and 48 h post peroxynitrite exposure. Peroxynitrite significantly increased transferase mediated deoxyuridine triphosphate-biotin nick end-labeling-positive neurons at all time points examined (P< or =0.001) compared with artificial cerebrospinal fluid controls (Two-way analysis of variance followed by Tukey test), peaking at 24 h post-exposure. Electron microscopic observation of characteristic features of apoptosis confirmed peroxynitrite-induced neuronal apoptosis. Total transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling-positive cells were counted in areas near and 0.2 mm away from the fiber track. The counts both peaked at 24 h with no significant difference between the two areas. However, at 6 and 12 h post-exposure the counts were significantly higher near than away from the fiber track (P=0.03 and P=0.007 respectively, paired t test). Immunohistochemical staining indicates caspase-3 was activated by peroxynitrite; this activation peaked at 6 h post-exposure, suggesting that activation of caspase-3 might be an early event in the apoptotic cell death cascade. We conclude that 1) peroxynitrite generated in the cord at the level produced by spinal cord injury induces neuronal apoptosis, indicating a role for peroxynitrite in secondary spinal cord injury; 2) caspase activation might be involved in peroxynitrite-induced neuronal apoptosis; 3) therefore removal of peroxynitrite should reduce secondary cell death after spinal cord injury. PMID- 12535939 TI - Sex differences in mouse cortical thickness are independent of the complement of sex chromosomes. AB - Although the morphology of the cerebral cortex is known to be sexually dimorphic in several species, to date this difference has not been investigated in mice. The present study is the first to report that the mouse cerebral cortex is thicker in males than in females. We further asked if this sex difference is the result of gonadal hormones, or alternatively is induced by a direct effect of genes encoded on the sex chromosomes. The traditional view of mammalian neural sexual differentiation is that androgens or their metabolites act during early development to masculinize the brain, whereas a feminine brain develops in the relative absence of sex steroids. We used mice in which the testis determination gene Sry was inherited independently from the rest of the Y chromosome to produce XX animals that possessed either ovaries or testes, and XY animals that possessed either testes or ovaries. Thus, the design allowed assessment of the role of sex chromosome genes, independent of gonadal hormones, in the ontogeny of sex differences in the mouse cerebral cortex. When a sex difference was present, mice possessing testes were invariably masculine in the morphology of the cerebral cortex, independent of the complement of their sex chromosomes (XX vs. XY), and mice with ovaries always displayed the feminine phenotype. These data suggest that sex differences in cortical thickness are under the control of gonadal steroids and not sex chromosomal complement. However, it is unclear whether it is the presence of testicular secretions or the absence of ovarian hormones that is responsible for the thicker male cerebral cortex. PMID- 12535940 TI - Methyl-CpG-binding protein 2 is localized in the postsynaptic compartment: an immunochemical study of subcellular fractions. AB - Methyl-CpG-binding protein 2 is a characteristic member of the methyl-CpG-binding protein family of transcription regulators. In conjunction with Sin3, MeCP2 recruits class I histone deacetylases to methyl-CpG regions to suppress transcription. Rett syndrome, a disorder characterized by mental retardation and autistic features, is associated in a majority of cases with mutations within the coding region of the MeCP2 gene. Considering that defective MeCP2 has mainly been related to Rett syndrome and other neurologic manifestations, we examined methyl CpG-binding protein 2 cellular and subcellular compartmentalization in normal brain by immunochemical methods. Methyl-CpG-binding protein 2 immunoreactivity is present mainly in neurons; while the few immunostained glia show label confined to nuclei, many neurons also show slight perikaryal staining. Using well characterized tissue fractions, we found that methyl-CpG-binding protein 2 but not Sin3 is found in both nuclear and postsynaptic compartments. This novel extranuclear localization is not unique to methyl-CpG-binding protein 2, since it has been previously reported for other transcription regulators such as c-Fos. These findings support the concept that methyl-CpG-binding protein 2 may link synaptic activity and transcriptional regulation in neurons. PMID- 12535941 TI - Modulation of spinal nociceptive processing through the glutamate transporter GLT 1. AB - GLT-1 is the predominant glutamate transporter in most brain regions and therefore plays a major role in terminating synaptic transmission and protecting neurons from glutamate neurotoxicity. In the present study we assessed (i) the regulation of GLT-1 expression in the spinal cord after peripheral nociceptive stimulation and (ii) the nociceptive behavior of rats following inhibition or transient knockdown of spinal GLT-1. Formalin injection into one hindpaw caused a rapid transient upregulation of GLT-1 protein expression in the spinal cord which did not occur when rats were pretreated with morphine (10 mg/kg, i.p.) suggesting that the nociceptive input specifically caused the increase of GLT-1 transcription. Inhibition of GLT-1 by the transportable inhibitor trans pyrrolidine-2,4-dicarboxylic acid resulted in a significant reduction of nociceptive behavior in the rat formalin assay. Similar results were obtained with a transient reduction of GLT-1 protein expression by antisense oligonucleotides. These data suggest that inhibition of GLT-1 activity or expression reduces excitatory synaptic efficacy and thereby nociception. Mechanisms that might explain this phenomenon may include activation of inhibitory metabotropic glutamate receptors, postsynaptic desensitization or disturbance of glutamate recycling. PMID- 12535942 TI - Suppression of the morphine-induced rewarding effect and G-protein activation in the lower midbrain following nerve injury in the mouse: involvement of G-protein coupled receptor kinase 2. AB - The present study was designed to investigate whether a state of neuropathic pain induced by sciatic nerve ligation could alter the rewarding effect, antinociception, and G-protein activation induced by a prototype of mu-opioid receptor agonist morphine in the mouse. The sciatic nerve ligation caused a long lasting and profound thermal hyperalgesia. Under this neuropathic pain-like state, an i.c.v. morphine-induced place preference was observed in sham-operated mice but not in sciatic nerve-ligated mice. However, no differences in the antinociceptive effect of i.c.v.-administered morphine were noted between the groups. The increases in the binding of guanosine-5'-o-(3 [(35)S]thio)triphosphate induced by morphine in lower midbrain membranes including the ventral tegmental area, which contributes to the expression of the rewarding effect of opioid, were significantly attenuated in sciatic nerve ligated mice. On the other hand, there were no differences in the stimulation of guanosine-5'-o-(3-[(35)S]thio)triphosphate binding to pons/medulla membranes, which plays an important role in the antinociception of mu-opioid receptor agonists, between the groups. In addition, no changes in levels of guanosine-5'-o (3-[(35)S]thio)triphosphate binding by either the selective delta- or kappa opioid receptor agonists were noted in membrane of the lower midbrain and limbic forebrain membranes obtained from sciatic nerve-ligated mice. Reverse transcription-polymerase chain reaction analysis showed that sciatic nerve ligation did not alter the mRNA product of mu-opioid receptors in the lower midbrain, indicating that a decrease in some mu-opioid receptor functions may result from the uncoupling of mu-opioid receptors from G-proteins. We found a significant increase in protein levels of G-protein-coupled receptor kinase 2, which causes receptor phosphorylation in membranes of the lower midbrain but not in the pons/medulla, obtained from mice with nerve injury, whereas there were no changes in the protein level of phosphorylated-protein kinase C in the lower midbrain. These results suggest that the uncoupling of mu-opioid receptors from G proteins by G-protein-coupled receptor kinase 2 in the lower midbrain may, at least in part, contribute to the suppression of the rewarding effect of morphine under neuropathic pain. PMID- 12535943 TI - Presynaptic inhibition of synaptic transmission by adenosine in rat subthalamic nucleus in vitro. AB - Whole-cell patch clamp recordings were made from the subthalamic nucleus in rat brain slice preparations to examine the effect of adenosine on inhibitory and excitatory synaptic transmission. Adenosine reversibly inhibited both GABA mediated inhibitory and glutamate-mediated excitatory postsynaptic currents. Adenosine at 100 microM reduced the amplitude of inhibitory and excitatory postsynaptic currents by 42+/-5% and 34+/-6%, respectively. Reductions in the amplitude of both inhibitory and excitatory postsynaptic currents were accompanied by increases in paired-pulse ratios. In addition, adenosine decreased the frequency of spontaneous miniature excitatory postsynaptic currents but had no effect on their amplitude. These results are consistent with a presynaptic site of action. The adenosine A(1) receptor antagonist 8-cyclopentyl-1,3 dipropylxanthine completely reversed the adenosine-induced attenuation of inhibitory and excitatory postsynaptic currents, but 8-cyclopentyl-1,3 dipropylxanthine alone had no effect on synaptic currents evoked at 0.1 Hz. However, 8-cyclopentyl-1,3-dipropylxanthine inhibited a time-dependent depression of excitatory postsynaptic currents that was normally observed in response to a 5 Hz train of stimuli, suggesting that endogenous adenosine could be released during higher frequencies of stimulation. These results suggest that adenosine inhibits synaptic release of GABA and glutamate by stimulation of presynaptic A(1) receptors in the subthalamic nucleus. PMID- 12535944 TI - Ultrastructural localization of serotonin2A receptors in the middle layers of the rat prelimbic prefrontal cortex. AB - Cortical serotonin(2A) receptors are hypothesized to be involved in the pathology and treatment of schizophrenia. Light microscopic studies in the rat prefrontal cortex have localized serotonin(2A) receptors to the dendritic shafts of pyramidal and local circuit neurons. Electrophysiological studies have predicted that these receptors are also located on glutamate terminals, whereas neurochemical studies have hypothesized that they are located on dopamine terminals in this area. The present study sought to determine the ultrastructural localization of immunoperoxidase labeling for serotonin(2A) receptors in the middle layers of the prelimbic portion of the rat prefrontal cortex. Serotonin(2A) receptor immunoreactivity was observed in 325 identifiable structures. Of these, 73% were postsynaptic profiles that were composed of either dendritic shafts (58%) or dendritic spine heads and necks (42%). Twenty-four percent of the labeled profiles were presynaptic axons and varicosities; most of these had morphological features that were characteristic of monoamine axons: thin diameter, lack of myelination, occasional content of dense-cored vesicles, and infrequent formation of synapses in single sections. The remainder of the labeled profiles (4%) were glial processes. These findings suggest that serotonin(2A) receptor-mediated effects within the rat prelimbic prefrontal cortex are primarily postsynaptic in nature, affecting both the spines of pyramidal cells and the dendrites of pyramidal and local circuit neurons in this area. The results further suggest that serotonin acts presynaptically via this receptor subtype, most likely at receptors on monoamine fibers, and only rarely directly on glutamate axons. PMID- 12535945 TI - Intracerebroventricular injection of a neuropeptide Y-Y1 receptor agonist increases while BIBP3226, a Y1 antagonist, reduces the infarct volume following transient middle cerebral artery occlusion in rats. AB - Recent studies using middle cerebral artery occlusion in the rat have suggested a role of neuropeptide Y in ischemic pathophysiology. In this study, we investigated the effects of an i.c.v. injection of a neuropeptide Y-Y2 receptor agonist, neuropeptide Y 3-36, a Y1 receptor agonist, [Leu(31),Pro(34)] neuropeptide Y, or a Y1 receptor antagonist, BIBP3226, on infarct volume and hemodynamic parameters following middle cerebral artery occlusion. Adult male Sprague-Dawley rats were subjected to transient middle cerebral artery occlusion for 2 h. A single i.c.v. injection of neuropeptide Y 3-36 (15 microg/kg), [Leu(31),Pro(34)]-neuropeptide Y (30 microg/kg), or BIBP3226 (5, 15, or 45 microg/kg) was given at 30 min of ischemia. Blood pressure, heart rate, and regional cerebral perfusion were monitored during ischemia and reperfusion. The rats were decapitated after 70 h of reperfusion, and their brains were cut into 2 mm-thick coronal slices before reaction with a 2% solution of 2,3,5 triphenyltetrazolium chloride to reveal the infarct. When compared with an infarct volume of 17.4+/-4.4% of the ipsilateral hemisphere following injection of neuropeptide Y 3-36, administration of the Y1 receptor analogs significantly modified the infarct volume (ordinary one-way analysis of variance (ANOVA), P<0.0001). [Leu(31),Pro(34)]-neuropeptide Y increased the infarct volume to 32.0+/-4.1% (Student-Newman-Keuls post-test, P<0.01), whereas BIBP3226 at 15 microg/kg decreased the infarct volume to 6.5+/-1.0% (post-test P<0.05). Although there was no major difference in the hemodynamic parameters among the groups, injection of [Leu(31),Pro(34)]-neuropeptide Y tended to further reduce cerebral perfusion during ischemia, while injection of BIBP3226 at 15 microg/kg appeared to have the opposite effect. In addition to glutamate, calcium ion and nitric oxide, activation of the neuropeptide Y-Y1 receptors may mediate cerebral damage during focal ischemia. Conversely, inhibiting the Y1 receptors may protect the brain against ischemic injury. Further studies are warranted to confirm the neuroprotective potential of neuropeptide Y-Y1 receptor inhibition. PMID- 12535946 TI - Catechol O-methyltransferase mRNA expression in human and rat brain: evidence for a role in cortical neuronal function. AB - Catechol O-methyltransferase (COMT) is involved in the inactivation of catecholamines, including the neurotransmitter dopamine. A Val(108/158) Met functional polymorphism of the COMT gene has been shown to affect working memory associated frontal lobe function in humans. In the present study, in situ hybridization histochemistry was employed to determine the mRNA expression profile of COMT in the human prefrontal cortex, striatum and midbrain and in the rat forebrain. In both species, COMT mRNA signals were observed in large pyramidal and smaller neurons in all cortical layers of the prefrontal cortex as well as in medium and large neurons in the striatum. Levels of COMT mRNA were obviously higher in neurons than in glia. The striatum, which receives a dense dopaminergic input, expressed lower levels of COMT mRNA as compared with the prefrontal cortex. Consistent with previous protein expression data, COMT mRNA was abundant in ependymal cells lining the cerebral ventricles. In the midbrain, COMT mRNA was detected in dopaminergic neurons in both species, albeit at low levels. In the rat forebrain, dense labeling was also detected in choroid plexus and hippocampal dentate gyrus and Ammon's horn neurons. Contrary to expectations that COMT would be expressed predominantly in non-neuronal cells, the present study shows that neurons are the main cell populations expressing COMT mRNA in the prefrontal cortex and striatum. Combined with previous data about protein localization, the present results suggest that the membrane-bound isoform of COMT having a high affinity for dopamine is expressed at neuronal dendritic processes in human cortex, consistent with functional evidence that it plays an important role in dopaminergic neurotransmission. PMID- 12535948 TI - Distribution of gephyrin in the human brain: an immunohistochemical analysis. AB - Gephyrin is an ubiquitously expressed protein that, in the central nervous system, generates a protein scaffold to anchor inhibitory neurotransmitter receptors in the postsynaptic membrane. It was first identified as a protein component of the glycine receptor complex. Recent studies have demonstrated that gephyrin is colocalized with several subtypes of GABA(A) receptors and is part of postsynaptic GABA(A) receptor clusters. Here, we describe a study of the regional and cellular distribution of gephyrin in the human brain, determined by immunohistochemical localisation at the light and confocal laser scanning microscopic levels. At the regional level, gephyrin immunoreactivity was observed in most of the major brain regions examined. The most intense staining was in the cerebral cortex, hippocampus and caudate-putamen, in various brainstem nuclei with more moderate levels in the thalamus and cerebellum. At the cellular level gephyrin immunoreactivity was present on the plasma membranes of the soma and dendrites of pyramidal neurons throughout the various cortical regions examined. In the hippocampus, intense staining was observed on the granule cells of the dentate gyrus, and neurons of the CA1 and CA3 regions showed intense punctate gephyrin staining on their apical dendrites and cell bodies. Gephyrin immunoreactivity was also observed on neurons in the thalamus, globus pallidus and substantia nigra. In the putamen intense labelling of the striosomes was observed; most of the medium-sized neurons in the caudate-putamen were weakly labelled and many large neurons of the striatum were conspicuously stained. Many of the brainstem nuclei, notably the dorsal motor nucleus of the vagus, hypoglossal nucleus, trigeminal nucleus and inferior olive were all labelled with gephyrin. The spinal cord also showed high levels of gephyrin immunoreactivity. Our results demonstrate that the anchoring protein gephyrin is ubiquitously present in the human brain. We therefore suggest that gephyrin may have a central organizer role in assembling and stabilizing inhibitory postsynaptic membranes in human brain and is similar in function to those observed in the rodent brain. These findings contribute towards elucidating the role of gephyrin in the human brain. PMID- 12535947 TI - ERK1/2 activation in rat ventral tegmental area by the mu-opioid agonist fentanyl: an in vitro study. AB - Opioid receptors in the ventral tegmental area, predominantly the mu-opioid receptors, have been suggested to modulate reinforcement sensitivity for both opioid and non-opioid drugs of abuse. The present study was conducted to study signal transduction proteins, which may mediate the functioning of mu-opioid receptors in the neurons of the ventral tegmental area. Therefore, brain slices of the ventral tegmental area were exposed in vitro to the specific mu-opioid agonist fentanyl and immunohistochemically stained for four different activated proteins using phospho-specific antibodies. Fentanyl dose-dependently activated extracellular signal-regulated protein in brain slices of the ventral tegmental area. This activation was reversible with naloxone. Furthermore, naloxone itself also activated extracellular signal-regulated protein kinase. Under the present conditions fentanyl did not affect extracellular signal-regulated protein kinase 1 and 2, Stat and cyclic AMP-response element-binding protein activity. The direct activation of extracellular signal-regulated protein kinase in ventral tegmental area slices by the mu-opioid agonist fentanyl may suggest a role of extracellular signal-regulated protein kinase in reward processes. PMID- 12535949 TI - Excitation by dopamine of rat subthalamic nucleus neurones in vitro-a direct action with unconventional pharmacology. AB - Recent anatomical and physiological studies have pointed to a functional innervation of the subthalamic nucleus by dopamine. This nucleus has a pivotal role in basal ganglia function and voluntary movement control and the possibility that dopamine, and dopaminergic medication used in Parkinson's disease, might directly influence its activity is of considerable interest. We have evaluated electrophysiologically the action and pharmacology of dopamine on single subthalamic neurones in rat brain slices. Dopamine increased firing rate to up to a mean of 60% in 98% of the 261 neurones tested when examined using extracellular single-unit recording. This excitation was unaffected by the GABA antagonist picrotoxin, and the glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3 dione, and persisted in a low Ca(2+)/raised Mg(2+) solution, indicative of a direct action, independent of synaptic transmission. Of the 33 cells examined using whole patch-clamp recording, only 13 showed measurable increases in firing rate and/or depolarisations in response to dopamine. Dopamine-responsive cells displayed significantly greater access resistance, suggesting that an unidentified cytoplamic constituent, removed by whole-cell dialysis, was required for the response. Using extracellular recording, the D2-like dopamine receptor agonists quinpirole and bromocryptine, but not the D1-like receptor agonist 1 phenyl-2,3,4,5-tetrahydro-(1H)-3-benzazepine-7,8-diol, also consistently caused an excitation. This was mimicked by the catecholamine releaser amphetamine in 60% of cells tested. However, the dopamine excitation was not significantly reduced either by the D1-like receptor antagonist 7-chloro8-hydroxy-3-methyl-1-phenyl 2,3,4,5-tetrahydro-1H-3-benzazepine or the D2-like receptor antagonists (-) sulpiride, eticlopride and (+)-butaclamol, and the quinpirole excitation was also unaffected by (-)-sulpiride. In contrast, (-)-sulpiride, eticlopride and (+) butaclamol all abolished the D2-like receptor-mediated inhibition by dopamine of substantia nigra pars compacta neurones. The alpha-adrenoceptor antagonist phentolamine was a weak antagonist of dopamine excitations, but not of those caused by quinpirole. Dopamine excitations also showed weak sensitivity to the 5 HT(2) antagonist ritanserin, but were unaffected by the alpha(1)-adrenoceptor antagonist prazocin and the beta-adrenoceptor antagonist propranolol. The pharmacology of this dopamine excitation is inconsistent with an action on any known catecholamine receptor. However, the effect of amphetamine indicates that an unidentified monamine--possibly dopamine--can be released within the subthalamic nucleus to cause an excitation. The anomalies of its pharmacological characterisation do not strongly support a physiologically relevant direct action of dopamine in the rat subthalamic nucleus. PMID- 12535950 TI - Activation of nitric oxide-producing neurons in the brain stem during cardiac sympathoexcitatory reflexes in the cat. AB - Our previous studies have shown that selective inhibition of nitric oxide in the brain reduces pressor responses to activation of cardiac sympathetic afferents, thus suggesting that nitric oxide is involved in central regulation of cardiac cardiovascular sympathoexcitatory reflexes. Central neural regions in which nitric oxide-producing neurons are activated during these reflexes have not been well characterized. In the present study, we located nitric oxide-producing neurons in the brain stem activated by the input from cardiac sympathetic afferents by detecting colocalization of c-Fos immunoreactivity with nitric oxide synthesizing neurons. Expression of c-Fos has been used as a marker of activated neurons. Nitric oxide-producing neurons were identified by histochemical labeling of nicotine adenine dinucleotide phosphate-diaphorase (NADPH-d). In anesthetized cats with bilateral barodenervation and cervical vagotomy, bradykinin (1-10 microg in 0.1 ml; n=6) was applied to the anterior surface of the left ventricle six times every 20 minutes. Repetitive application of bradykinin consistently increased blood pressure, while the vehicle for bradykinin (0.9% saline, n=5) produced no responses. A substantial fraction (6-27%) of NADPH-d positive neurons displayed Fos immunoreactivity in the nucleus of the solitary tract, caudal and rostral ventral lateral medulla, lateral tegmental field, locus coeruleus and parabrachial nucleus in the bradykinin-treated cats. However, either no or rare (1-4%) double-labeled cells were found in these regions in control animals. Thus, nitric oxide-producing neurons are activated in several regions in the brain stem during stimulation of cardiac sympathetic afferents by bradykinin. Our data suggest that nitric oxide functions as a neurotransmitter/modulator in these areas to regulate the cardiac sympathoexcitatory reflexes. PMID- 12535951 TI - Blockade of epinephrine priming of the cerebral auditory evoked response by cortical cholinergic deafferentation. AB - The present study tested hypotheses derived from a neurobehavioral model of anxiety that posits an important role of the basal forebrain cholinergic system in the cortical processing of anxiety-associated stimuli and contexts. We hypothesized that visceral afferent activity induced by systemic administration of epinephrine would enhance the processing of auditory stimuli as evidenced by the cerebral auditory evoked response. We further predicted that selective lesions of the basal forebrain cortical cholinergic projection system would disrupt this processing, and would further block the effects of epinephrine. Results confirmed these hypotheses. Epinephrine was found to enhance the amplitude of the P70 component of the auditory evoked response in rats. Selective lesions of the basal forebrain corticopetal cholinergic projection, by intrabasalis infusions of 192 IgG saporin, delayed and reduced the amplitude of the P70 component, and blocked the potentiating effects of epinephrine on the auditory evoked response. The present results are consistent with the view that visceral afferent input may modulate cortical processing of sensory signals via the basal forebrain cholinergic system. These considerations emphasize the potential importance of ascending, bottom-up modulation of processing by telencephalic circuits that may impact on a wide range of behavioral functions. PMID- 12535952 TI - Differential increase in the expression of heat shock protein family members during sleep deprivation and during sleep. AB - Although sleep is thought to be restorative from prior wakeful activities, it is not clear what is being restored. To determine whether the synthesis of macromolecules is increased in the cerebral cortex during sleep, we subjected C57BL/6 mice to 6 hours of sleep deprivation and then screened the expression of 1176 genes of known function by using cDNA arrays. The expression of the heat shock proteins (HSP), endoplasmic reticulum protein (ERp72) and glucose-regulated protein (GRp78), was among the genes whose expression was significantly elevated in the cortex during sleep deprivation, whereas GRp78 and GRp94 mRNAs were elevated in the cortex during recovery sleep after sleep deprivation, as confirmed by conventional and quantitative real-time polymerase chain reaction and/or Northern analyses. A systematic evaluation of the expression of six heat shock protein family members (ERP72, GRp78, GRp94, HSP27, HSP70-1, and HSP84) in seven brain regions revealed increased mRNA levels in cortex, basal forebrain, hypothalamus, cerebellum and medulla during sleep deprivation, whereas increased mRNA levels during recovery sleep were limited to the cortex and medulla. Immunohistochemical studies identified increased numbers of GRp78-, GRp94-, and ERp72-immunoreactive cells in the dorsal and lateral cortex during sleep deprivation but, during recovery sleep, elevated numbers of these cells were found only in the lateral cortex. In the medulla, increased numbers of GRp94 immunoreactive cells were observed in nucleus tractus solitarius, dorsal motor nucleus of the vagus and the rostroventrolateral medulla during recovery sleep. The widespread increase of heat shock protein family mRNAs in brain during sleep deprivation may be a neuroprotective response to prolonged wakefulness. In contrast, the relatively limited heat shock protein family mRNA expression during recovery sleep may be related to the role of heat shock proteins in protein biogenesis and thus to the restorative function of sleep. PMID- 12535953 TI - Hippocampal network patterns of activity in the mouse. AB - Genetic engineering of the mouse brain allows investigators to address novel hypotheses in vivo. Because of the paucity of information on the network patterns of the mouse hippocampus, we investigated the electrical patterns in the behaving animal using multisite silicon probes and wire tetrodes. Theta (6-9 Hz) and gamma (40-100 Hz) oscillations were present during exploration and rapid eye movement sleep. Gamma power and theta power were comodulated and gamma power varied as a function of the theta cycle. Pyramidal cells and putative interneurons were phase locked to theta oscillations. During immobility, consummatory behaviors and slow wave sleep, sharp waves were present in cornu ammonis region CA1 of the hippocampus stratum radiatum associated with 140-200-Hz "ripples" in the pyramidal cell layer and population burst of CA1 neurons. In the hilus, large amplitude "dentate spikes" occurred in association with increased discharge of hilar neurons. The amplitude of field patterns was larger in the mouse than in the rat, likely reflecting the higher neuron density in a smaller brain. We suggest that the main hippocampal network patterns are mediated by similar pathways and mechanisms in mouse and rat. PMID- 12535954 TI - Dose- and sex-dependent effects of the neurotoxin 6-hydroxydopamine on the nigrostriatal dopaminergic pathway of adult rats: differential actions of estrogen in males and females. AB - Epidemiological and clinical studies provide growing evidence for marked sex differences in the incidence of certain neurological disorders that are largely attributed to the neuroprotective effects of estrogen. Thus there is a keen interest in the clinical potential of estrogen-related compounds to act as novel therapeutic agents in conditions of neuronal injury and neurodegeneration such as Parkinson's disease. Studies employing animal models of neurodegeneration in ovariectomised female rats treated with estrogen support this hypothesis, yet experimental evidence for sex differences in the CNS response to direct neurotoxic insult is limited and, as yet, few studies have addressed the role played by endogenously produced hormones in neuroprotection. Therefore, in this study we aimed to determine (1) whether the prevailing levels of sex steroid hormones in the intact rat provide a degree of protection against neuronal assault in females compared with males and (2) whether sex differences depend solely on male/female differences in circulating estrogen levels or whether androgens could also play a role. Using the selective, centrally administered neurotoxin 6-hydroxydopamine, which induces a lesion in the nigrostriatal dopaminergic pathway similar to that seen in Parkinson's disease, we have demonstrated a sexually dimorphic (male-dominant), dose-dependent susceptibility in rats. Furthermore, following gonadectomy, dopamine depletion resulting from a submaximal dose of 6-hydroxydopamine (1 microg) was reduced in male rats, whereas in females, ovariectomy enhanced dopamine depletion. Administration of the nonaromatizable androgen dihydrotestosterone to gonadectomized animals had no significant effect on 6-hydroxydopamine toxicity in either males or females, whereas treatment of gonadectomized males and females with physiological levels of estrogen restored the extent of striatal dopamine loss to that seen in intact rats, viz, estrogen therapy reduced lesion size in females but increased it in males. Taken together, our findings strongly suggest that there are sex differences in the mechanisms whereby nigrostriatal dopaminergic neurones respond to injury. They also reveal that the reported clinically beneficial effects of estrogen in females may not be universally adopted for males. While the reasons for this gender-determined difference in response to the activational action of estrogen are unknown, we hypothesize that they may well be related to the early organizational events mediated by sex steroid hormones, which ultimately result in the sexual differentiation of the brain. PMID- 12535955 TI - Effects of lateral hypothalamic lesion with the neurotoxin hypocretin-2-saporin on sleep in Long-Evans rats. AB - Narcolepsy, a disabling neurological disorder characterized by excessive daytime sleepiness, sleep attacks, sleep fragmentation, cataplexy, sleep-onset rapid eye movement sleep periods and hypnagogic hallucinations was recently linked to a loss of neurons containing the neuropeptide hypocretin. There is considerable variability in the severity of symptoms between narcoleptic patients, which could be related to the extent of neuronal loss in the lateral hypothalamus. To investigate this possibility, we administered two concentrations (90 ng or 490 ng in a volume of 0.5 microl) of the neurotoxin hypocretin-2-saporin, unconjugated saporin or saline directly to the lateral hypothalamus and monitored sleep, the entrained and free-running rhythm of core body temperature and activity. Neurons stained for hypocretin or for the neuronal specific marker were counted in the perifornical area, dorsomedial and ventromedial nucleus of the hypothalamus. More neuronal nuclei (NeuN) cells were destroyed by the higher concentration of hypocretin-2-saporin (-55%) compared with the lower concentration (-34%) in the perifornical area, although both concentrations lesioned the hypocretin neurons almost equally well (high concentration=91%; low concentration=88%). The high concentration of hypocretin-2-saporin also lesioned neurons in the dorsomedial nucleus of the hypothalamus and ventromedial nucleus of the hypothalamus. Narcoleptic-like sleep behavior was produced by both concentrations of the hypocretin-2-saporin. The high concentration produced a larger increase in non rapid eye movement sleep amounts during the normally active night cycle than low concentration. Neither concentration of hypocretin-2-saporin disrupted the phase or period of the core temperature or activity rhythms. The low concentration of unconjugated saporin did not significantly lesion hypocretin or neurons and did not alter sleep. The high concentration of unconjugated saporin produced some loss of neuronal nuclei-immunoreactive (NeuN-ir) neurons and hypocretin immunoreactive neurons, but only a transient increase in non-rapid eye movement sleep. These results led us to conclude that the extent of hypocretin neuronal loss together with an accompanying loss of cells in the lateral hypothalamus may explain the differences in severity of symptoms seen in human narcolepsy. PMID- 12535956 TI - Lack of vesicular zinc in mossy fibers does not affect synaptic excitability of CA3 pyramidal cells in zinc transporter 3 knockout mice. AB - Zinc is found throughout the CNS in synaptic vesicles of glutamatergic neurons and has been suggested to have a modulatory role in the brain because of its interaction with voltage- and ligand-gated ion channels. We took advantage of zinc transporter 3 knockout mice, which lack vesicular zinc, to study the possible physiological role of this heavy metal in hippocampal mossy fiber neurotransmission. We examined postsynaptic responses evoked by mossy fiber activation, recorded in CA3 pyramidal cells in hippocampal slices prepared from zinc transporter 3 knockout and wild-type mice. Field-potential response threshold and amplitude, input-output curves, and paired-pulse evoked responses were the same in slices from zinc transporter 3 knockout and wild-type mice. Furthermore, neither amplitude nor duration of pharmacologically isolated N methyl-D-aspartate, non-N-methyl-D-aspartate, GABA(A), and GABA(B) receptor mediated postsynaptic potentials differed between zinc transporter 3 knockout and wild-type mice. There was no difference in the magnitude of epileptiform discharges evoked by repetitive stimulation or kainic acid application. However, in slices from zinc transporter 3 knockout mice, there was greater attenuation of GABA(A)-mediated inhibitory postsynaptic potentials during tetanic stimulation compared with slices from wild-type animals. We conclude that lack of vesicular zinc in mossy fibers does not significantly affect the mossy fiber-associated synaptic excitability of CA3 pyramidal cells; however, zinc may modulate GABAergic synaptic transmission under conditions of intensive activation. PMID- 12535957 TI - Suppression of genioglossus muscle tone and activity during reflex hypercapnic stimulation by GABA(A) mechanisms at the hypoglossal motor nucleus in vivo. AB - The genioglossus muscle is involved in the maintenance of an open airway for effective breathing. Inhibitory neurotransmitters may be responsible for the major suppression of hypoglossal motor output to genioglossus muscle that occurs in certain behaviours such as rapid-eye-movement sleep. There is evidence for GABA(A) receptor-mediated inhibition of hypoglossal motoneurons in vitro. However, comparable studies have not been performed in vivo and the interactions of such mechanisms with integrative reflex respiratory control have also not been determined. Urethane-anaesthetised, tracheotomized and vagotomized rats were studied whilst diaphragm and genioglossus muscle activities, blood pressure and the electroencephalogram were recorded. Microdialysis probes were implanted into the hypoglossal motor nucleus, with sites verified by histology. Genioglossus responses to microdialysis perfusion of muscimol (GABA(A) agonist: 0, 0.1, 1 and 10 microM in artificial cerebrospinal fluid) were recorded at inspired CO(2)s of 0, 5 and 7.5% in six rats. Responses to bicuculline (GABA(A) antagonist, 0, 1, 10, 100 and 1000 microM) were also studied in six rats with and without CO(2) stimulation. Genioglossus activity decreased with muscimol (P<0.0001), with major suppression at 1 and 10 microM during air breathing (decreases=70.2% and 92.8%, P<0.005). Genioglossus activity increased with CO(2) (P=0.003), but genioglossus activation with 5 and 7.5% CO(2) were almost abolished with 10-microM muscimol. Responses were specific to genioglossus muscle as there were no changes in diaphragm, respiratory rate or blood pressure with muscimol (P>0.144). Antagonism of GABA(A) receptors increased genioglossus activity (P<0.001). These results show that GABA(A) receptor stimulation at the hypoglossal motor nucleus suppresses both genioglossus muscle tone and activity in the presence of reflex stimulation produced by hypercapnia. Recruitment of such mechanisms may contribute to the major suppression of genioglossus activity observed with and without CO(2) stimulation in behaviours such as rapid-eye-movement sleep. PMID- 12535958 TI - Glial cell line neurotrophic factor-family receptor alpha-1 is present in central neurons with distinct phenotypes. AB - Glial cell line neurotrophic factor(GDNF) is a potent survival factor for several types of neurons. GDNF binds with high affinity to the GDNF-family receptor alpha 1 (GFRalpha-1) which is expressed in different brain areas. In the present study, by using anatomical techniques, we document the phenotypic diversity among GFRalpha-1 expressing neurons in the CNS. GFRalpha-1 expression was found in GABA (gamma-aminobutyric acid)-containing neurons distributed in the cortex, reticular thalamic nucleus and septum. While high expression of GFRalpha-1 was often observed in cholinergic motoneurons in the spinal cord, very few septal cholinergic neurons were found to express GFRalpha-1. GFRalpha-1 transcripts were also detected in catecholaminergic neurons in the periventricular hypothalamic nucleus, dorsal raphe nucleus and locus ceruleus. Within the raphe nucleus, GFRalpha-1 expression was prominent in many serotonergic neurons and in few neurons containing the enzyme nitric oxide synthase. As GFRalpha-1 is activated by GDNF and GDNF-related neurotrophic factors, the widespread distribution of GFRalpha-1 in neurons with different phenotypes indicates that the neuronal activity of these neurons is likely to be affected by GDNF and GDNF-related neurotrophic factors. This would result in the regulation of diverse neuronal pathways in the adult brain. Published by Elsevier Science Ltd on behalf of IBRO. PMID- 12535959 TI - Single administration of interleukin-1 increased corticotropin releasing hormone and corticotropin releasing hormone-receptor mRNA in the hypothalamic paraventricular nucleus which paralleled long-lasting (weeks) sensitization to emotional stressors. AB - Single exposure to the proinflammatory cytokine interleukin-1 induces sensitization of the adrenocorticotropin hormone and corticosterone responses to stressors weeks later (hypothalamus-pituitary-adrenal sensitization). Hypothalamus-pituitary-adrenal responses are controlled by corticotropin releasing hormone and arginine-vasopressin secreted from parvocellular corticotropin-releasing hormone neurons of the hypothalamic paraventricular nucleus and may involve autoexcitatory feedback mechanisms. Therefore, we studied the temporal relationship between resting levels of corticotropin-releasing hormone, corticotropin-releasing hormone-R1 and arginine-vasopressin receptor (V1a, V1b) mRNAs in the paraventricular nucleus and the development of hypothalamus-pituitary-adrenal sensitization to an emotional stressor (novelty). The adrenocorticotropin hormone precursor molecule proopiomelanocortin hnRNA in the pituitary gland served as an index for acute activation. Single administration of interleukin-1 induced sensitization of the hypothalamus pituitary-adrenal to novelty from 3 to 22 days later, but not after 42 days. Single administration of interleukin-1 induced biphasic increases in corticotropin-releasing hormone and corticotropin-releasing hormone-R1 mRNAs in the paraventricular nucleus: an early peak within 24 h, followed by a delayed (>7 days) increase that peaked after 22 days. Hypothalamic V1a and V1b mRNA levels were unaffected. In contrast, in the pituitary gland, there was an early decrease in corticotropin-releasing hormone-R1 mRNA (from 10.5 to 3 h after interleukin-1) and V1b receptor mRNA (3 to 6 h), which returned to control levels from 24 h onwards. Thus, interleukin-1-induced long-lasting hypothalamus-pituitary-adrenal sensitizations associated with prolonged activation of corticotropin-releasing hormone and corticotropin-releasing hormone-R1 mRNA expression in the paraventricular nucleus, but not with changes in the expression of proopiomelanocortin hnRNA or V1b receptor or corticotropin-releasing hormone R1 mRNAs in the pituitary gland. We propose that transient exposure to immune events can induce long-lasting hypothalamus-pituitary-adrenal sensitization, which at least in part involves long-term hypothalamic adaptations that enhance central corticotropin-releasing hormone signaling. PMID- 12535960 TI - Effects of basolateral amygdala dopamine depletion on the nucleus accumbens and medial prefrontal cortical dopamine responses to stress. AB - In vivo voltammetry was used to study the effects of basolateral amygdala dopamine depletion on stress-induced dopamine release in the nucleus accumbens and medial prefrontal cortex. Male Long-Evans rats received bilateral microinjections of 6-hydroxydopamine or vehicle into the basolateral amygdala. Changes in dopamine signal were monitored in the nucleus accumbens and in the right and left hemispheres of medial prefrontal cortex, in lesioned animals and shams. Animals were subjected to a physical stressor (tail pinch) and a species typical threat (fox odour); each stressor was presented twice over four consecutive daily sessions. The results indicate that the nucleus accumbens dopamine responses to both stressors are significantly potentiated by dopamine depleting lesions to basolateral amygdala. In contrast, while the dopamine stress response in the left medial prefrontal cortex did not differ between lesioned animals and shams, the right medial prefrontal cortical dopamine response to tail pinch, but not fox odour stress, was significantly attenuated in lesioned animals. Therefore, basolateral amygdala dopamine depletion had opposite effects on the nucleus accumbens and medial prefrontal cortical dopamine responses to stress, although the effect on the latter is lateralized to the right hemisphere in a stressor-specific manner. These data indicate that stress-induced activation of meso-amygdaloid dopamine exerts an inhibitory influence on the nucleus accumbens dopamine response to stress. They also suggest the possibility that meso-amygdaloid dopamine influences the nucleus accumbens dopamine response to stress indirectly by modulating stress-induced dopamine release in medial prefrontal cortex. These findings add to a growing body of evidence of a preferential involvement of right medial prefrontal cortical dopamine in a wide range of physiological responses to stress. PMID- 12535961 TI - Independent modulation of basal and feeding-evoked dopamine efflux in the nucleus accumbens and medial prefrontal cortex by the central and basolateral amygdalar nuclei in the rat. AB - Interactions of the central and basolateral nuclei of the amygdala with the mesocorticolimbic dopamine system are implicated in the acquisition and performance of conditioned responses for food reward. This study investigated whether dopamine transmission in the nucleus accumbens and the medial prefrontal cortex of the rat is influenced by the amygdala and if so, to assess the significance of the interaction in free feeding of a palatable food. To this end, we examined the effects of reverse-dialysis of the sodium channel blocker lidocaine into either the central or basolateral on dopamine efflux in the nucleus accumbens and the medial prefrontal cortex as determined by microdialysis and high-pressure liquid chromatography with electrochemical detection. The present results revealed for the first time that inactivation of the central decreased basal levels of dopamine efflux in the nucleus accumbens, but not in the medial prefrontal cortex. Furthermore, administration of lidocaine into the central significantly attenuated feeding-evoked increases in dopamine efflux in both terminal regions. These neurochemical effects were accompanied by feeding related behaviours akin to the Kluver-Bucy syndrome. In contrast, inactivation of the basolateral affected neither food intake nor dopamine efflux in the nucleus accumbens, but triggered dramatic long-lasting oscillations in dopamine efflux in the medial prefrontal cortex, irrespective of whether food was presented or not. Overall, these findings indicate that the central and basolateral independently modulate dopamine transmission in both terminal regions of the mesocorticolimbic dopamine system. The central, in particular, and its influence on the dopamine system, may be involved in the regulation of food intake. PMID- 12535962 TI - Effects of oligonucleotide antisense to dopamine D3 receptor mRNA in a rodent model of behavioural sensitization to levodopa. AB - Levodopa-induced dyskinesias are abnormal involuntary movements that develop as a side-effect of long-term treatment with levodopa for Parkinson's disease. The mechanisms underlying such effects are unclear but may include abnormal stimulation of dopamine D(3) receptors. Elevations in dopamine D(3) receptor mRNA and binding are seen in the denervated striatum of hemiparkinsonian rats treated chronically with levodopa, and these changes correlate well with behavioural sensitization in this model. Further investigation of dopamine D(3) receptor involvement in levodopa-induced dyskinesias is hampered by the lack of appropriately selective ligands for this receptor. Here, in vivo administration of an antisense oligonucleotide designed to reduce striatal dopamine D(3) receptor expression provides a level of specificity not available through traditional pharmacological approaches. Following chronic treatment with levodopa, hemiparkinsonian rats received intrastriatal infusion of oligonucleotide antisense to dopamine D(3) receptor mRNA for 5 days. Antisense treatment effectively and selectively reduced striatal dopamine D(3) receptor binding and blocked behavioural sensitization to the effects of repeated levodopa. These findings confirm the importance of the D3 receptor in the expression of behavioural sensitization to levodopa in animals with dopaminergic denervation and contribute to our limited understanding of the functional significance of this receptor. In that sensitization to the effects of repeated levodopa in this setting may be analogous to medication-induced dyskinesias in humans, our findings furthermore suggest that drugs which block D(3) function may be helpful in the treatment of dyskinesias, without necessarily exacerbating Parkinsonism. PMID- 12535963 TI - Environmental carpetbaggers. PMID- 12535964 TI - The cold-water coral Lophelia pertusa (Scleractinia) and enigmatic seabed mounds along the north-east Atlantic margin: are they related? AB - In this study, an updated distribution of Lophelia pertusa between the Porcupine Seabight and Norwegian shelf is presented. It seems unlikely that enigmatic mound structures observed at water depths of more than 570 m during acoustic seabed surveys, particularly to the west of the Shetland Islands, are related to the occurrence of L. pertusa. At these depths in the Faroe-Shetland Channel, the predominant influence of cold Arctic water precludes its growth. Iceberg dumpsites are also considered unlikely explanations for the origin of these mounds, and they are interpreted as most likely to be related to the release of fluids at the seabed. When mound structures were investigated, no scleractinian corals were recovered at water depths >500 m. This study shows the importance of seabed temperature as an environmental control on cold-water coral distribution. The significance of cold-water coral habitats in sustaining high levels of local scale biodiversity is now becoming apparent in parallel with increased hydrocarbon extraction and fishing activity beyond the shelf edge. There is growing evidence that these areas have been marked by the passage of deep-water trawls. It seems likely that trawling activity has already reduced the extent of cold-water coral distribution in this region of the north-east Atlantic. PMID- 12535965 TI - The EU directive on port reception facilities for ship-generated waste and cargo residues: current availability of facilities in the North Sea. AB - The aim of this paper is to evaluate the current availability of port reception facilities within the North Sea area. The evaluation is based primarily on original survey data from the summer of 2001, supplemented by existing information from surveys conducted by the International Maritime Organization. The EU Directive on port reception facilities for ship-generated waste etc. (Directive 2000/59/EC) requires that all ports provide facilities to meet the needs of those vessels normally calling in at them. In order for the Directive to be implemented effectively, the physical availability of such facilities is vital. This paper audits the wide range of port types within the region and also outlines the wide range of vessel types using these ports, and their requirements for a range of facilities. On the basis of this analysis, conclusions are drawn about the extent to which the provision requirement of the Directive is already being met. PMID- 12535966 TI - Can we always ignore ship-generated food waste? AB - Considerable quantities of food waste can be generated at a rapid rate in ships, particularly those with large numbers of people onboard. By virtue of the amounts involved and its nature, food waste is potentially the most difficult to manage component of a ship's garbage stream, however, in most sea areas it may be dealt with by the simple expedient of direct discharge to sea. As a consequence, only minimal attention is paid to food waste management by many ship and port operators and advisory bodies, and there is a paucity of information in the available literature. The determination that management of ships' food waste is inconsequential is, however, incorrect in many circumstances. Disposal to sea is not always possible due to restrictions imposed by MARPOL 73/78 and other marine pollution control instruments. Effective management of food waste can be critical for ships that operate in areas where disposal is restricted or totally prohibited. PMID- 12535967 TI - The concept of "sustainable ecological succession"; and its value in assessing the recovery of sediment seabed biodiversity from environmental impact. AB - The problem of determining whether the biodiversity of an impacted muddy seabed is recovering can be resolved by drawing on a concept termed "sustainable ecological succession". At a site impacted by discharge of mine tailings, a suite of approximately 6 primary opportunist species (mostly polychaete worms) had started to sustain itself within 1-2 years after discharge ceased (1995), within the mix of 100+ other species which were not sustaining themselves. The start of a sustaining ecological succession is easily measurable by repeat surveys, and requires only the services of one taxonomic identifier to demonstrate the consistent presence and numbers of a limited range of species. At the assessed site, by 2000, some secondary opportunist species had entered the succession, and the species richness of the impacted area had come to equal that of the reference areas. PMID- 12535968 TI - Recovery simulations of grossly polluted sediments in the Bilbao Estuary. AB - The Bilbao Estuary is one of the most contaminated estuaries on the north coast of Spain, and vast efforts have been made to abate pollution there. In fact, the local water authority has forecast a biological recovery of the native fauna after a substantial increase in dissolved oxygen to normoxic levels. In order to assess this prediction by evaluating the extent of natural regeneration of these polluted sediments, two long-term bioassays (t=90 d) were performed. In both of them, lethal (differences in survival) and sublethal (differences in length and weight growth) effects were measured by using juvenile individuals of the autochthonous clam, Scrobicularia plana (Da Costa, 1778). The sediments tested differed in pollution levels, as measured by a set of indicators including PAHs, PCBs, heavy metals, volatile organic matter and coprostanol. All sediments were finally exposed to normoxic conditions in situ in the Bilbao Estuary (DO approximately 6.3 mg l(-1)). In the first experiment, concerning moderately polluted sediments from the Bilbao Estuary and reference sediments from the "pristine" Plentzia Estuary, no significant differences (P>0.05) were found regarding animal survival (approximately 94.5%) or growth in length or weight between the sediments tested. In the second experiment, also involving grossly polluted sediments (GPS) from the Bilbao Estuary, survival (24.5%) was statistically lower (P<0.05) than in the other sediments (approximately 93%). No significant differences were found in growth (length, weight) between animals exposed to moderately polluted or reference sediments. We interpret this dramatic difference in survival as the lethal effect on the animals tested of the GPS of the Bilbao Estuary, indicating a situation where biological recovery is not possible due to the adverse consequences of contaminants sorbed into sediments. The extensive use of this inexpensive bioassay could help to distinguish sediments in which homeostatic recovery is possible from grossly polluted "hot spots", which need costly remedial actions. PMID- 12535969 TI - Spills of fuel oil #6 and Orimulsion can have indistinguishable effects on the benthic meiofauna. AB - Fuel oil #6 is used for the production of electrical power in the United States. Orimulsion is being considered as an alternative fuel, but its value and risk compared to fuel oil #6 need to be assessed. Our study examined the relative impact of accidental spills of the two hydrocarbons on the meiofaunal community. To do so, we maintained microcosms of the shallow, sandy, subtidal environment for three months. Treatment microcosms received a single application of hydrocarbon-coated sand. As indicators of effect, we used copepod and nematode abundance and copepod species diversity, sex ratio, fecundity, age structure, and neutral-lipid content. A comparison of the hydrocarbon treatments showed no significant differences. The tests had adequate power to detect ecologically significant changes. Our results indicate that a spill of Orimulsion would have approximately the same impact as a spill of fuel oil #6 on the meiofauna. PMID- 12535970 TI - A toxicity identification evaluation of silty marine harbor sediments to characterize persistent and non-persistent constituents. AB - Sediment toxicity in silty marine harbor sediments is frequently dominated by ammonia or sulfide, leaving the adverse effects of persistent toxic substances unnoticed. To investigate the latter, we subjected interstitial water from three contaminated silty sediments to toxicity identification evaluation (TIE) phase I manipulations and tested for toxicity with four bioassays: the amphipod Corophium volutator (survival as an endpoint), the sea urchin Psammechinus miliaris (fertilization, embryo development) and the bacterium Vibrio fischeri (bioluminescence inhibition). The graduated pH manipulations identified the prominent toxicity of ammonia in the amphipod and sea urchin embryo tests, and also sulfide toxicity in the bacterium test. In two of the three samples tested with the amphipods, sea urchin embryos and bacteria, a small but significant reduction in interstitial water toxicity was achieved by removing persistent compounds through C(18) solid phase extraction. EDTA chelation resulted in a slight detoxification of the interstitial water for the amphipods and sea urchin embryos, but this was not related to any measured trace metals. Despite the presence of toxic levels of ammonia and sulfide in the harbor sediments, we established the adverse biological effects of persistent constituents by means of the TIE manipulations and in vivo interstitial water bioassays. PMID- 12535971 TI - Survivorship and growth of Fucus gardneri after transplant to an acid mine drainage-polluted area. AB - Acid mine drainage (AMD) from an abandoned copper mine at Britannia Beach, British Columbia, Canada, enters the marine environment through Britannia Creek. The surrounding intertidal zone is devoid of rockweed, Fucus gardneri Silva, a seaweed that dominates nearby shores. Rockweed plants were transplanted to the intertidal zone near Britannia Creek and monitored for changes in percent cover, survivorship, growth rate and Cu content. Autumn and winter transplants to within 100 m of Britannia Creek resulted in negative growth rates and high mortality within 57 days of exposure to AMD, with Cu levels in rockweed surpassing 2,300 ppm in dry tissue. Summer transplants to sites 300-700 m from Britannia Creek showed no consistent differences between AMD-exposed rockweed and control plants, possibly because the plants were stressed by desiccation. The results are consistent with ecological effects observed in other studies, and provide strong evidence for the role of AMD in excluding rockweed from the shores near Britannia Creek. PMID- 12535972 TI - Emergent effects of heavy metal pollution at a population level: Littorina brevicula a study case. AB - Changes in genetic variability and allele frequency can be responses from natural populations when encountering a novel contaminated environment. The genetic diversity and population structuring of natural populations of the gastropod Littorina brevicula from heavy-metal polluted and unpolluted environments along the southeast coast of Korea were examined using two mtDNA markers, cyt b and ND6. This study applied a nested clade analysis to test the existence of structuring association of haplotype distribution with environments (polluted and unpolluted). No genetic differences within cyt b mtDNA were detected between environments. On the other hand, differences in population haplotype diversity and structuring were found within ND6 mtDNA between polluted and unpolluted environments. The ND6-mtDNA haplotype (=genetic) diversity was significant lower in polluted environments. This decreased genetic diversity along with differences in the haplotype distribution within heavy-metal polluted environments compared to those unpolluted ones stand out as emergent effects from pollution at a population level. In this study, we propose the use of different approaches, such as the NCA, that takes into account the rare haplotypes, when assessing the effects of pollution on population genetic structuring. PMID- 12535973 TI - A field survey of chemicals and biological products used in shrimp farming. AB - This study documented the use of chemicals and biological products in marine and brackish water shrimp farming in Thailand, the world's top producer of farmed shrimp. Interviews were conducted with 76 shrimp farmers in three major shrimp producing regions, the eastern Gulf coast, the southern Gulf coast and the Andaman coast area. Farmers in the study used on average 13 different chemicals and biological products. The most commonly used products were soil and water treatment products, pesticides and disinfectants. Farmers in the southern Gulf coast area used a larger number of products than farmers in the other two areas. In the study, the use of more than 290 different chemicals and biological products was documented. Many of the pesticides, disinfectants and antibiotics used by the farmers could have negative effects on the cultured shrimps, cause a risk for food safety, occupational health, and/or have negative effects on adjacent ecosystems. Manufacturers and retailers of the products often neglected to provide farmers with necessary information regarding active ingredient and relevant instructions for safe and efficient use. PMID- 12535974 TI - Identifying potential marine pests--a deductive approach applied to Australia. AB - Introduced marine organisms continue to threaten coastal resources around the world. Many agencies rely on lists of potentially harmful species for risk assessment and to prioritise management responses to an incursion. This study outlines a deductive hazard assessment technique to identify potential marine pests that may arrive via ballast water and/or hull fouling. This technique is then applied in an Australian context to identify potential "next pests" for this region. An extensive literature review identified 851 introduced marine species from around the world. The following selection criteria were then applied to develop a next pest list: (a) the species has been reported in a shipping vector or has a ship-mediated invasion history; (b) the vector still exists; (c) the species is responsible for economic or environmental harm; and, (d) it is exotic to Australia or present in Australia but subject to official control. The selection criteria are transparent and consistent with other international and national biosecurity initiatives. Thirty three of the species identified in the initial literature review satisfied all four selection criteria. These species are described here together with their associated vectors and impacts. PMID- 12535975 TI - Acoustic characteristics of fish bombing: potential to develop an automated blast detector. AB - The use of explosives to catch fish has caused extensive damage to coral reefs throughout Southeast Asia, but the frequency with which they are used is largely unknown. The aim of this work is to develop a detection system capable of distinguishing underwater explosions from background noise, and locating their origin by triangulation. Blast signals have been recorded over a range of distances and the key features that differentiate them from background noise have been determined. For small charges the effective range of such a detector is more than 12 km and may extend up to 50 km depending on the mass of the charges being used. Such a system would help to determine the scale of the problem, identify areas at greatest risk and quantify the effectiveness of management intervention designed to control destructive fishing practices. It may assist with fisheries enforcement in some areas. PMID- 12535976 TI - Cytochrome P4501A expression, chemical contaminants and histopathology in roach, goby and sturgeon and chemical contaminants in sediments from the Caspian Sea, Lake Balkhash and the Ily River Delta, Kazakhstan. AB - Roach, goby and sturgeon were examined for cytochrome P4501A (CYP1A) expression and histopathology, in relation to contaminant burdens in fish and sediment. Gradients of induction of CYP1A were observed. Roach from the Ural and Ily River Deltas and roach and goby from the two stations nearest the Caspian Sea oil fields displayed higher levels of CYP1A expression in several organs than was observed in fish from further offshore. Great sturgeon and Russian sturgeon showed higher levels of CYP1A expression than was seen in starred sturgeon and gobies in the Ural delta. No fish showed evidence of contaminant-related histopathologies in the organs examined, despite the elevated CYP1A levels. Low levels of polychlorinated biphenyls and elevated levels of inshore and riverine petroleum hydrocarbons from these habitats suggest that this ongoing hydrocarbon exposure, and that from natural sources and long-term oil exploration on the Northeastern Caspian shore, contributed to the CYP1A induction observed. PMID- 12535977 TI - Electrokinetic potentials of mineral oxides and calcium carbonate in artificial seawater. AB - Electroacoustic method was used to measure the zeta potentials of mineral oxides and calcium carbonate in artificial sea water (pH 8), and the following values were obtained: synthetic hematite: 0.87 mV; goethite: 1.01 mV; fumed silica: 3.56 mV; quartz: -1.38 mV; calcium carbonate: 1.91 mV. The absolute values of the zeta potentials obtained in the present study are substantially lower than the zeta potentials in seawater reported by other authors. Our results indicate that the effect of electrostatic repulsion in the interactions between the particles of these materials in seawater is negligible. PMID- 12535978 TI - The distributions of particulate heavy metals and its indication to the transfer of sediments in the Changjiang Estuary and Hangzhou Bay, China. AB - Particulate heavy metals (Cu, Pb, Cd) were measured following intensive sampling in June and October 1994 at 70 stations in the Changjiang Estuary and Hangzhou Bay, China. In the study area, particulate Cu and Pb have a similar level that is higher than the concentration of particulate Cd. Cu, Pb and Cd concentrations in suspended sediments are higher than those in bed load. In the middle of Hangzhou Bay, heavy metal concentrations in suspended sediments and bed load are similar. This may be the result of the frequent exchange between them, which is due to the strong dynamic condition. A negative relationship was observed between concentrations of heavy metals in bed load and bulk density. Fine-grained sediments were the main carriers of heavy metals. Sedimentary dynamics dominate the fate of heavy metals in these sites. The distributions of metals (especially Cu) in suspended sediments can indicate the transfer of sediments in this area. PMID- 12535979 TI - Quantification and classification of marine litter on the municipal beach of Ensenada, Baja California, Mexico. PMID- 12535980 TI - Trace metals in ribbed mussels from Arthur Kill, New York/New Jersey, USA. PMID- 12535981 TI - Trace metals in Fanga'uta Lagoon, Kingdom of Tonga. PMID- 12535982 TI - Cone pigment variations in four genera of new world monkeys. AB - Previous research revealed significant individual variations in opsin genes and cone photopigments in several species of platyrrhine (New World) monkeys and showed that these in turn can yield significant variations in color vision. To extend the understanding of the nature of color vision in New World monkeys, electroretinogram flicker photometry was used to obtain spectral sensitivity measurements from representatives of four platyrrhine genera (Cebus, Leontopithecus, Saguinus, Pithecia). Animals from each genus were found to be polymorphic for middle to long-wavelength (M/L) sensitive cones. The presence of a short-wavelength sensitive photopigment was established as well so these animals conform to the earlier pattern in predicting that all male monkeys are dichromats while, depending on their opsin gene array, individual females can be either dichromatic or trichromatic. Across subjects a total of five different M/L cone pigments were inferred with a subset of three of these present in each species. PMID- 12535983 TI - Dexanabinol (HU-211) has a beneficial effect on axonal sprouting and survival after rat optic nerve crush injury. AB - Dexanabinol (HU-211) is a synthetic non-psychotropic cannabinoid and a non competitive NMDA-receptor antagonist. The beneficial effect of dexanabinol on prevention of degeneration and promotion of regeneration was studied on the crush injured rat optic nerve model. Sprague-Dawley rats were subjected to a calibrated crush injury of the optic nerve and treated with a single intraperitoneal injection of dexanabinol (7 mg/kg), its vehicle only or were untreated. Transmission electron microscopic analysis of the excised optic nerves was performed after 30 days. In the dexanabinol treated rats, the site of injury was traversed by unmyelinated and thinly myelinated axons, possibly indicative of regenerative growth. No such growth was detectable in the controls. Viable axons were found 0.5 mm distal to the site of injury in 6 of 8 dexanabinol treated rats, but in only 1 of 10 rats in the control groups. These results have clinical implications for the prevention of secondary degeneration and promotion of regeneration after injuries to the central nervous system. PMID- 12535984 TI - Sensitivity to contrast modulation: the spatial frequency dependence of second order vision. AB - We consider the overall shape of the second-order modulation sensitivity function (MSF). Because second-order modulations of local contrast or orientation require a carrier signal, it is necessary to evaluate modulation sensitivity against a variety of carriers before reaching a general conclusion about second-order sensitivity. Here we present second-order sensitivity functions for new carrier types (low pass (1/f) noise, and high pass noise) and demonstrate that, when first-order artefacts have been accounted for, the shape of the resulting MSFs are similar to one another and to those for white and broad band noise. They are all low pass with a likely upper frequency limit in the range 10-20 c/deg, suggesting that detection of second-order stimuli is relatively insensitive to the structure of the carrier signal. This result contrasts strongly with that found for (first-order) luminance modulations of the same noise types. Here the noise acts as mask and each noise type masks most those frequencies that are dominant in its spectrum. Thus the shape of second-order MSFs are largely independent of the spectrum of their noise carrier, but first-order CSFs depend on the spectrum of an additive noise mask. This provides further evidence for the separation of first- and second-order vision and characterises second-order vision as a low pass mechanism. PMID- 12535985 TI - Temporal asymmetry in motion masking: a shortening of the temporal impulse response function. AB - Morgan and Chubb observed a striking temporal asymmetry in motion masking (Vis. Res. 39 (1999) 4217). Motion was produced with a two-frame sequence of gratings presented in spatial quadrature phase; the second grating (100 ms) was presented immediately after the first grating (100 ms), with no temporal overlap. The contrast threshold for detecting the direction of motion of the stimulus pair was facilitated when the first grating was of low-contrast and the second grating was of high-contrast, but strong masking occurred when the order was reversed, so the high-contrast grating came first. We replicated this result, but showed that the masking mostly disappeared when the two gratings temporally overlapped only slightly. The high sensitivity to the precise temporal pattern of the stimulus can be explained by a small temporal 'shortening' of the temporal impulse response function (IRF) as stimulus contrast is increased. The IRF is biphasic with a negative inhibitory lobe. When the first grating has high-contrast, its flash response (owing to the shortening of the IRF) may be in a fairly strong negative phase by the time that the positive response to the second, lower contrast grating has reached appreciable strength--this reduces the magnitude of the motion signal generated by the two flashes and can account for the masking. A shortening of the IRF with increased contrast (a nonlinearity) is supported by psychophysical studies in humans and by recordings of magnocellular retinal ganglion cells in macaque, and the present results bolster this concept. PMID- 12535986 TI - Simulation of artificial vision: I. Eccentric reading of isolated words, and perceptual learning. AB - Simulations of artificial vision were performed to assess "minimum requirements for useful artificial vision". Retinal prostheses will be implanted at a fixed (and probably eccentric) location of the retina. To mimic this condition on normal observers, we projected stimuli of various sizes and content on a defined stabilised area of the visual field. In experiment 1, we asked subjects to read isolated 4-letter words presented at various degrees of pixelisation and at various eccentricities. Reading performance dropped abruptly when the number of pixels was reduced below a certain threshold. For central reading, a viewing area containing about 300 pixels was necessary for close to perfect reading (>90% correctly read words). At eccentricities beyond 10 degrees, close to perfect reading was never achieved even if more than 300 pixels were used. A control experiment using isolated letter recognition in the same conditions suggested that lower reading performance at high eccentricity was in part due to the "crowding effect". In experiment 2, we investigated whether the task of eccentric reading under such specific conditions could be improved by training. Two subjects, naive to this task, were trained to read pixelised 4-letter words presented at 15 degrees eccentricity. Reading performance of both subjects increased impressively throughout the experiment. Low initial reading scores (range 6%-23% correct) improved impressively (range 64%-85% correct) after about one month of training (about 1 h/day). Control tests demonstrated that the learning process consisted essentially in an adaptation to use an eccentric area of the retina for reading. These results indicate that functional retinal implants consisting of more than 300 stimulation contacts will be needed. They might successfully restore some reading abilities in blind patients, even if they have to be placed outside the foveal area. Reaching optimal performance may, however, require a significant adaptation process. PMID- 12535987 TI - Human temporal impulse response speeds up with increased stimulus contrast. AB - It is well known that raising mean luminance speeds-up the visual response to temporal change. At higher mean luminance, the temporal impulse response function (IRF) becomes more transient or biphasic. An analogous effect is observed physiologically when stimulus contrast is increased, at constant mean luminance. As stimulus contrast is raised, the temporal response to flicker advances in phase and becomes more transient (bandpass). The MC (magnocellular) retinal ganglion cells manifest this temporal contrast gain control, but the PC (parvocellular) cells do not. We show psychophysically that the temporal response in humans speeds-up in an analogous manner as stimulus contrast is raised. Low spatial-frequency gratings, of suprathreshold contrast, were presented as pairs of pulses, separated by brief delays. Responses became more transient with increasing contrast in both our motion task (direction discrimination) and in our flicker task ('agitation' discrimination), mimicking the temporal contrast gain control seen in the physiological studies. Results could be modeled with a nonlinearity, in which the IRF shortens with increasing contrast. PMID- 12535988 TI - Visual evoked potentials to line- and luminance-defined triangles. AB - Processing of line- and luminance-defined triangles was investigated by studying visual potentials (VEPs) evoked by triangles. Twenty-six subjects were randomly presented with line, grey, and illusory triangles. Relative to VEPs elicited by grey and illusory triangles, VEPs to line triangles included P220s that were smaller at frontal sites but larger at occipital sites, and N260s that were smaller over both temporal and occipital sites. It is proposed that, in contrast to triangle defined by a line, illusory and luminance-defined triangles include information involved in the processing of surface. PMID- 12535989 TI - The influence of cyclovergence on unconstrained stereoscopic matching. AB - In order to perceive depth from binocular disparities the visual system has to identify matching features of the two retinal images. Normally, the assigned disparity is unambiguously determined by monocularly visible matching constraints. The assigned disparity is ambiguous when matching is unconstrained, such as when we view an isolated long oblique disparate line. Recently we found that in order to perceive a depth probe at the same depth as the oblique line, the probe needs to have the same horizontal disparity as the line (i.e. matching occurs along horizontal "search-zones" [Vis. Res. 40 (2000) 151]). Here we examined whether the depth probe disparity in unconstrained matching of long lines is influenced by cyclovergence, by cyclorotation between stereogram half images, or by combinations of the two. We measured retinal rotation (>6 deg in cyclovergence conditions). We found that in those conditions in which the retinal images were the same (a condition with, say, both zero cyclovergence and zero cyclorotation between the half-images, creates the same retinal images as a condition with both 6 deg cyclovergence and 6 deg cyclorotation) assigned depth was the same too, i.e. independent of cyclovergence. Thus, the assigned depth of the test-line seems to be determined solely by the retinal test-line orientation, implying that the binocular matching algorithm does not seem to incorporate the eyes' cyclovergence when matching is unconstrained. PMID- 12535990 TI - Comparison of performance on memory-guided saccade and delayed spatial match-to sample tasks in monkeys. AB - To investigate the sources of spatial error in memory-guided saccades (MGS), we have trained monkeys on two different tasks: a MGS task and a delayed spatial match-to-sample (MTS) task. We first tested the effect of introducing a post saccadic visual feedback on the accuracy of MGS. We found that visual feedback had a pronounced effect on the systematic saccade error, but less of an effect on the variable error. Visual feedback can improve the accuracy of saccadic eye movements over several days, while feedback removal can decrease accuracy in a reversible way. These effects also depend both on target eccentricity and the duration of the memory delay. To test whether saccade error is due to the accuracy of spatial memory storage or arises downstream from that memory, we measured behavioral performance on a spatial MTS task both before and after training with visual feedback. The results showed no significant difference in performance of the MTS task before and after feedback training despite significant changes in MGS accuracy. The results suggest that the accuracy of spatial memory is not the source of the systematic errors that accompany MGS. PMID- 12535991 TI - Oculomotor strategies for the direction of gaze tested with a real-world activity. AB - Laboratory-based models of oculomotor strategy that differ in the amount and type of top-down information were evaluated against a baseline case of random scanning for predicting the gaze patterns of subjects performing a real-world activity- walking to a target. Images of four subjects' eyes and field of view were simultaneously recorded as they performed the mobility task. Offline analyses generated movies of the eye on scene and a categorization scheme was used to classify the locations of the fixations. Frames from each subject's eye-on-scene movie served as input to the models, and the location of each model's predicted fixations was classified using the same categorization scheme. The results showed that models with no top-down information (visual salience model) or with only coarse feature information performed no better than a random scanner; the models' ordered fixation locations (gaze pattern) matched less than a quarter of the subjects' gaze patterns. A model that used only geographic information outperformed the random scanner and matched approximately a third of the gaze patterns. The best performance was obtained from an oculomotor strategy that used both coarse feature and geographic information, matching nearly half the gaze patterns (48%). Thus, a model that uses top-down information about a target's coarse features and general vicinity does a fairly good job predicting fixation behavior, but it does not fully specify the gaze pattern of a subject walking to a target. Additional information is required, perhaps in the form of finer feature information or knowledge of a task's procedure. PMID- 12535992 TI - Disorders of vertical optokinetic nystagmus in patients with ocular misalignment. AB - PURPOSE: To compare responses to vertical and horizontal optokinetic (OK) stimulation in patients with disorders of ocular alignment. METHODS: Using the magnetic search coil technique, we measured horizontal and vertical rotations of both eyes in six patients with strabismus since childhood and eight normal subjects. The OK stimulus subtended 72 degrees horizontally and 60 degrees vertically, consisted of black-and-white stripes with a spatial frequency of 0.04 cycles/degree, and moved either vertically or horizontally at 22.5 or 12 degrees/s. All patients and controls were tested with both eyes viewing and monocularly. RESULTS: Vertical OK responses were asymmetric in most normals and patients. The direction of this asymmetry varied between individuals, but upward stimuli more commonly elicited a greater response than downward stimuli. Monocular horizontal OK responses were symmetric in normals; patients showed either an asymmetry with greater responses for nasal motion, or a directional bias. During monocular and binocular viewing, vertical OK stimulation induced vertical nystagmus in normal subjects, but all patients showed diagonal responses, with horizontal components that were significantly greater than controls. The inappropriate horizontal component of the response increased at the higher stimulus speed, and was not simply due to latent nystagmus. CONCLUSIONS: Patients with disorders of ocular alignment since childhood show an inappropriate horizontal response to vertical OK stimuli, indicating directional abnormality of either motion vision pathways or the ocular motor response. PMID- 12535993 TI - Differential ambiguity reduces grouping of metastable objects. AB - Two coaxial, ambiguously rotating objects tend to be perceived as corotating. Such grouping could be the consequence of bottom-up, cooperative interactions between the stimuli, or the top-down selection of object properties consistent with a model of the objects or scene. However, we find that the coupling between an ambiguous and unambiguous object is sharply reduced, presenting a challenge for both explanations of grouping. We describe experiments that support the idea that top-down feedback is necessary to select and stabilize a perceptual interpretation for ambiguous figures. Reduced coupling between objects of differing ambiguity can be explained if the feedback is global and proportional to ambiguity. PMID- 12535994 TI - Motion detection and the coincidence of structure at high and low spatial frequencies. AB - We used filtered random dot kinematograms and natural images to examine how motion detection depends the relative locations of structures defined at low and high spatial frequencies. The upper displacement limit of motion (D(max)), the lower displacement limit (D(min)) and motion coherence thresholds were unaffected by the degree of spatial coincidence between high and low spatial frequency structures i.e. whether they were consistent or inconsistent with a single feature. However motion detection was possible between band-pass filtered random dot patterns whose peak frequencies were separated by up to 4 octaves. The first result implicates spatial frequency selective motion detectors that operate independently. The second result implicates a motion system that can integrate the displacements of edges defined by widely separated spatial frequencies. Both are required to account for the two results, and they appear to operate under very similar conditions. PMID- 12535995 TI - Poor visibility of motion in depth is due to early motion averaging. AB - Under a variety of conditions, motion in depth from binocular cues is harder to detect than lateral motion in the frontoparallel plane. This is surprising, as the nasal-temporal motion in the left eye associated with motion in depth is easily detectable, as is the nasal-temporal motion in the right eye. It is only when the two motions are combined in binocular viewing that detection can become difficult. We previously suggested that the visibility of motion-in-depth is low because early stereomotion detectors average left and right retinal motions. For motion in depth, a neural averaging process would produce a motion signal close to zero. Here we tested the averaging hypothesis further. Specifically we asked, could the reduced visibility observed in previous experiments be associated with depth and layout in the stimuli, rather than motion averaging? We used anti correlated random dot stereograms to show that, despite no depth being perceived, it is still harder to detect motion when it is presented in opposite directions in the two eyes than when motion is presented in the same direction in the two eyes. This suggests that the motion in depth signal is lost due to early motion averaging, rather than due to the presence of noise from the perceived depth patterns in the stimulus. PMID- 12535996 TI - Representation of statistical properties. AB - Everyday scenes often contain sets of similar objects. Perceptual representations may summarize these with statistical descriptors. After determining the psychological mean of two sizes, we measured thresholds for judging the mean with arrays of 12 circles of heterogeneous sizes. They were close to those for the size of elements in homogeneous arrays and single elements, and were little affected by either exposure duration (50-1000 ms) or memory delays (up to 2s). They were only slightly more accurate within the same distribution than across different distributions (normal, uniform, two-peaks, and homogeneous), confirming that subjects were indeed averaging sizes. PMID- 12535997 TI - Correlation between hand preference and intraocular pressure from right- and left eyes in right- and left-handers. AB - To test whether there is a relationship between handedness and the intraocular pressure and there is a lateralization in the intraocular pressure, the intraocular pressures of the right- and left-eyes were compared in right- and left-handed students. The intraocular pressures were higher in the right-eye than in the left-eye in men, right-handers, and right-eyed subjects; there was no right-left difference in females, left-handers, left-eyed, and both-eyed subjects. The intraocular pressure of right- and left-eyes was higher in left handers than right-handers. And there were significant negative correlations between hand preference and the intraocular pressure of both right- and left eyes. We have concluded that the dominant eye has higher intraocular pressure compared to the nondominant one, and there is a relationship between hand preference and the intraocular pressure. PMID- 12535998 TI - Coordinating one hand with two eyes: optimizing for field of view in a pointing task. AB - We previously found that subjects switched 'ocular dominance' as a function of horizontal gaze direction in a reaching task [Vision Res. 41 (14) (2001) 1743]. Here we extend these findings to show that when subjects pointed to targets across the horizontal binocular field, they aligned the fingertip with a vertical plane located between the eyes and the target. This eye-target plane gradually shifted from aligning with the left eye (leftward targets) to between the two eyes (intermediate targets) to the right eye (rightward targets). We suggest that this occurs to optimize eye-hand alignment towards the eye with the best overall field of view. PMID- 12535999 TI - An evaluation of the lag of accommodation using photorefraction. AB - OBJECTIVE: The lag of accommodation which occurs in most human subjects during reading has been proposed to explain the association between reading and myopia. However, the measured lags are variable among different published studies and current knowledge on its magnitude rests largely on measurements with the Canon R 1 autorefractor. Therefore, we have measured it with another technique, eccentric infrared photorefraction (the PowerRefractor), and studied how it can be modified. METHODS: Particular care was taken to ensure correct calibration of the instrument. Ten young adult subjects were refracted both in the fixation axis of the right eye and from the midline between both eyes, while they read text both monocularly and binocularly at 1.5, 2, 3, 4 and 5 D distance ("group 1"). A second group of 10 subjects ("group 2"), measured from the midline between both eyes, was studied to analyze the effects of binocular vs monocular vision, addition of +1 or +2 D lenses, and of letter size. Spherical equivalents (SE) were analyzed in all cases. RESULTS: The lag of accommodation was variable among subjects (standard deviations among groups and viewing distances ranging from 0.18 to 1.07 D) but was significant when the measurements were done in the fixation axis (0.35 D at 3 D target distance to 0.60 D at 5 D with binocular vision; p<0.01 or better all cases). Refracting from the midline between both eyes tended to underestimate the lag of accommodation although this was significant only at 5 D (ANOVA: p<0.0001, post hoc t-test: p<0.05). There was a small improvement in accommodation precision with binocular compared to monocular viewing but significance was reached only for the 5 D reading target (group 1- lags for a 3/4/5 D target: 0.35 vs 0.41 D/0.48 vs 0.47 D/0.60 vs 0.66 D, ANOVA: p<0.0001, post hoc t-test: p<0.05; group 2--0.29 vs 0.12 D, 0.33 vs 0.16 D, 0.23 vs -0.31 D, ANOVA: p<0.0001, post hoc t-test: p<0.05). Adjusting the letter height for constant angular subtense (0.2 deg) induced scarcely more accommodation than keeping letter size constantly at 3.5 mm (ANOVA: p<0.0001, post hoc t-test: n.s.). Positive trial lenses reduced the lag of accommodation under monocular viewing conditions and even reversed it with binocular vision. CONCLUSIONS: After consideration of possible sources of measurement error, the lag of accommodation measured with photorefraction at 3 D (0.41 D SE monocular and 0.35 D SE binocular) was in the range of published values from the Canon R-1 autorefractor. With the measured lag, simulations of the retinal images for a diffraction limited eye suggest surprisingly poor letter contrast on the retina. PMID- 12536000 TI - Short-latency disparity-vergence eye movements in humans: sensitivity to simulated orthogonal tropias. AB - Small disparity stimuli applied to large random-dot patterns elicit machine-like vergence eye movements at short latency. We have examined the sensitivity of these eye movements to simulated orthogonal tropias in three normal subjects by recording (1) the effects of vertical disparities on the initial horizontal vergence responses elicited by 2 degrees crossed and uncrossed (horizontal) disparity stimuli, and (2) the effects of horizontal disparities on the initial vertical vergence responses elicited by 1.2 degrees left-hyper and 0.8 degrees right-hyper (vertical) disparity stimuli. Initial vergence responses were strongest when the orthogonal disparity was close to zero, and decreased to zero as the orthogonal disparity increased to 3 degrees -5 degrees, i.e., there was only a limited tolerance for orthogonal disparity. Tuning curves describing the dependence of the initial change in the vergence angle on the orthogonal disparity were well fit by a Gaussian function. An additional subject, who had an esotropia of approximately 10 degrees in our experimental setup, showed almost no horizontal vergence responses but did show vertical vergence responses to vertical disparity stimuli at short latency (albeit slightly longer than normal) despite the fact that her esotropia resulted in uncrossed disparities that would have totally disabled the vertical vergence mechanism of a normal subject, cf., anomalous retinal correspondence. PMID- 12536001 TI - Reading a population code: a multi-scale neural model for representing binocular disparity. AB - Although binocular neurons in the primary visual cortex are sensitive to retinal disparity, their activity does not constitute an unambiguous disparity signal. A multi-spatial-scale neural model for disparity computation is developed to examine how population activity might be interpreted to overcome ambiguities at the single neuron level. The model incorporates a front end that encodes disparity by a family of complex cell-like energy units and a second stage that reads the population activity. Disparity is recovered by matching the population response to a set of canonical templates, derived from the mean response to white noise stimuli at a range of disparities. Model predictions are qualitatively consistent with a variety of psychophysical results in the literature, including the effects of spatial frequency on stereoacuity and bias in perceived depths, and the effect of standing disparity on increment thresholds. Model predictions are also consistent with data on qualitative appearance of complex stimuli, including depth averaging, transparency, and corrugation. The model also accounts for the non-linear interaction of disparities in compound grating stimuli. These results show that a template-match approach reduces ambiguities in individual and pooled neuronal responses, and allows for a broader range of percepts, consistent with psychophysics, than other models. Thus, the pattern of neural population activity across spatial scales is a better candidate for the neural correlate of depth perception than the activity of single neurons or the pooled activity of multiple neurons. PMID- 12536002 TI - Contrast sensitivity in subgroups of developmental dyslexia. AB - It has been proposed that developmental dyslexia is associated with a deficit in the magnocellular pathway of the visual system. Other research focuses upon the heterogeneous nature of developmental dyslexia, and evidence that subgroups of dyslexia may be identified based on selective deficits in specific component reading skills. This study tested the hypothesis that visual processing deficits may be present in different subgroups of developmental dyslexia by comparing the visual contrast sensitivity of three subgroups of dyslexic children (phonological, surface and mixed) and controls. The stimulus designed to measure magnocellular visual function was a low spatial frequency Gaussian blob, flickered sinusoidally at a temporal frequency of 8.33 Hz. The control stimulus, designed to measure parvocellular visual function, was a relatively high spatial frequency Gaussian windowed grating (8 c/deg) slowly ramped on and off. There were no significant differences between the groups of dyslexic and control children in contrast sensitivity to either stimulus. The findings do not support the existence of a magnocellular system deficit in dyslexia. PMID- 12536003 TI - Mechanical properties of the rabbit iris smooth muscles. AB - The study focuses on obtaining the visco-elastic properties of the iris sphincter and dilator muscles. Two kinds of experiments were performed: the isometric contraction experiment and the isotonic quick release experiment. The length tension relationship was obtained from the former experiment. This relationship clarified the contribution of each muscle in determining the statics of the pupil. The viscous and serial elastic properties were obtained from the latter experiment. The viscosity could be expressed by the expanded Hill's equation as a function of velocity and contractile tension. We argue that serial elasticity is independent of contractile tension. These properties provide insights into the pupillary mechanism. PMID- 12536004 TI - Lactation and weaning effects on physiological and behavioral response to stressors. AB - Two experiments tested the effects of lactation and weaning on heart rate (HR), corticosterone, and behavioral responses to stress in Wistar rats. In Experiment 1, HRs in lactating, weaning, and control animals were recorded for 10-min periods before, during, and after immobilization stress. Compared with control animals, lactating and weaning animals showed a diminished initial HR response. In addition, HRs of weaning animals failed to habituate and showed a delayed decline after stress termination. In Experiment 2, behaviors, HRs, and corticosterone levels in the elevated plus maze (EPM) were compared among lactating, weaning, and control animals. Compared with control animals, weaning animals exhibited more anxiety behaviors. Contrary to expectation, compared to the other two groups, lactating animals exhibited more closed-arm entries, although they may have been motivated by maternal behavior, rather than anxiety. Initial HR responses to the plus maze were attenuated in lactating animals. Corticosterone levels after the plus maze were lowest in the lactating dams and highest in the control animals. The results from these two experiments are consistent with effects of breast-feeding and weaning observed in humans. In general, lactation is associated with an attenuated initial HR response to stress, while weaning is associated with exacerbated response to stressors. PMID- 12536005 TI - Energy intake and body composition in anorexia and bulimia nervosa. AB - Energy intake during the weight gain phase and the weight maintenance phase was examined in three groups of inpatients: 64 anorectic restrictors (AN-R), 37 anorectic bulimics (AN-B), and 74 normal-weight bulimics (BN). The influence of body composition and other variables such as weight, exercise, and bingeing and purging frequencies on energy intake was analyzed. Eating disorder subgroups were found to differ in energy intakes to gain weight and to maintain weight within a target weight range. Anorectic restrictors consumed significantly more energy to gain weight than did the anorectic bulimics (a mean of 3055 kcals per day vs. a mean of 2788 kcal per day). Energy intake for weight gain was inversely related to admission body mass index (BMI) and to admission fat-free mass. Bingeing and purging frequencies did not predict caloric intake for either weight gain or maintenance for anorectics. Energy intake of the anorectic patients when they were maintaining weight within a target weight range did not differ between restrictors and anorectic bulimics (means of 2204 and 2134 kcal/day, respectively). Energy needed to maintain weight within a target weight range was predicted by the amount of fat-free mass at target weight and by the mean number of calories ingested per week during weight gain. Bulimics consumed fewer calories than either of the two anorectic groups during the weight maintenance phase (1538 kcal/day). Other than BMI at hospital admission, bulimics' energy intakes were unrelated to all demographic and eating disorder-related variables, including body composition. PMID- 12536006 TI - Two effects of high-fat diets on the satiating potency of cholecystokinin-8. AB - Chronic ingestion of diets containing 34% or 54% fat have been reported [Peptides 19 (1998) 1407] to decrease the inhibitory effect on food intake of doses of cholecystokinin-8 (CCK-8) less than 1 microg/kg i.p. We attempted to replicate this phenomenon in three experiments by comparing the effect of high-fat and low fat diets on the threshold dose of CCK-8 for inhibition and on the dose-response function for doses of CCK-8 that ranged from 0.125 to 2.0 microg/kg. The first experiment tested rats five times per week. Rats on a 34% fat diet had a higher threshold (1.0 microg/kg) than rats on a 5% fat diet (0.25 microg/kg). The dose response functions, however, were not significantly different. This result replicated the earlier report [Peptides 19 (1998) 1407]. The second experiment tested rats maintained on the same diets every other day as in the original report. It failed, however, to replicate the results of the first experiment or the earlier report because the threshold doses and the dose-response functions of CCK-8 were not significantly different between rats on 34% and 5% fat diets. The third experiment compared the potency of CCK-8 in rats on a 60% fat diet with the potency in rats on a 5% fat diet. CCK-8 was significantly more potent in the rats on the 60% fat diet because the threshold dose of these rats was lower (0.125 microg/kg) than in rats on the 5% fat diet (0.25 microg/kg), and the dose response function in rats on the 60% fat diet was significantly more potent than in rats on the 5% fat diet. Thus, we observed two effects of the chronic ingestion of high-fat diets on the inhibitory potency of CCK-8: (1). A 34% fat diet increased the threshold dose of CCK-8 without changing the dose-response function in one of two experiments. (2). A 60% fat diet increased the potency of CCK-8 because it decreased the threshold dose and increased the dose-response function significantly. PMID- 12536007 TI - Effects of endurance training on the gluconeogenic capacity of periportal and perivenous hepatocytes. AB - This study was undertaken to assess the effect of 8 weeks of endurance training (treadmill, last 4 weeks: 60 min at 26 m/min, 8-10% grade) on the gluconeogenic capacity of periportal (PP-H) and perivenous (PV-H) hepatocytes of overnight fasted rats. Isolated PP-H and PV-H, obtained by selective destruction after liver perfusion with digitonin and collagenase, were incubated with saturating concentrations of a mixture of lactate and pyruvate (20:2 mM; Lac+Pyr) or alanine (20 mM; Ala) to determine the glucose production flux (J(glucose)) in the incubation medium. Endogenous J(glucose) as well as J(glucose) from substrates were significantly higher (P<.05) in PP-H than PV-H in the untrained state. Following training, a selective increase (P<.05) in J(glucose) from endogenous substrates and from Lac+Pyr was observed in PV-H only, resulting in the disappearance (P>.05) of the difference of J(glucose) between PP-H and PV-H. It is concluded that the increase in the gluconeogenic capacity of the liver following endurance training is first observed in PV-H. PMID- 12536008 TI - Self-injurious behavior in male rhesus macaques does not reflect externally directed aggression. AB - Self-injurious behaviors (SIB), such as self-biting and self-wounding, have been observed in a small percentage of captive nonhuman primates. Because rhesus monkeys that exhibit SIB also tend to be more aggressive, it was hypothesized that SIB is related to externally directed aggression and is associated with contexts in which physical contact between participants is prevented. The purpose of this study was to test the hypothesized relationship between SIB and outward aggression. Subjects were first presented with videotapes of conspecifics, scenery and a blank screen, and their behavior was recorded. Levels of salivary cortisol, an indicator of stress, were also measured before and after presentation of the videos. Although aggression increased when subjects viewed tapes containing conspecifics, neither cortisol levels nor self-biting behavior varied as a function of tape content. The subjects were then placed in two additional test situations: an empty room and the same room containing an unfamiliar conspecific. Aggression was significantly higher in the stranger condition compared to the empty room condition. The two situations yielded parallel increases in cortisol, suggesting that being alone was just as stressful as being paired with an unfamiliar conspecific. Self-biting rates were also similar in these two conditions. Thus, contrary to our prediction, increases in aggression did not correlate with increases in SIB. These results suggest that under similarly stressful conditions, SIB and externally directed aggression are unrelated. PMID- 12536009 TI - Food availability and photoperiod affect reproductive development and maintenance in the marsh rice rat (Oryzomys palustris). AB - To examine whether photoperiod and food availability interact to influence reproductive development (Experiment 1), we exposed juvenile male and female rice rats to 16L:8D or 14L:10D and to ad lib, 80% of ad lib, or 60% of ad lib food intake from 3 to 8 weeks of age and recorded body and reproductive organ masses. Absolute paired testis masses were similar in ad lib and 80% of ad lib groups but significantly different than the 60% of ad lib group in both photoperiods. Relative paired testis masses were significantly different in the 80% and 60% of ad lib groups on 16L:8D only. Absolute seminal vesicle masses (SVM) were directly dependent upon the level of food restriction in both photoperiods, but relative SVMs were different only in the 60% of ad lib group. Terminal body masses were also directly dependent upon the level of food restriction and were greater on 16L:8D than on 14L:10D at most levels of food availability. In juvenile females, absolute uterine mass was only affected in the 60% of ad lib group on 14L:10D, while absolute paired ovary masses were affected on both photoperiods in the 60% of ad lib groups only. There was no effect of photoperiod or food on relative uterine and paired ovary masses. Terminal body mass was affected by food intake in both photoperiods. Lastly, in adult males (Experiment 2), photoperiod and food restriction affected reproductive function. Within a photoperiod, there was no effect of food restriction (75% of ad lib) on the testes, seminal vesicles, or testosterone levels in animals housed on 16L:8D, but terminal body mass was significantly reduced. On 12L:12D, however, food restriction significantly decreased testes and SVMs relative to ad lib-fed controls. Testosterone levels were reduced regardless of food availability. There was no effect of food restriction on terminal body mass. These results suggest that multiple potential environmental cues can be utilized to affect gonadal status in both juvenile and adult marsh rice rats. PMID- 12536010 TI - Assessment of sucrose and ethanol reinforcement: the across-session breakpoint procedure. AB - We have demonstrated previously that the use of an across-session progressive ratio procedure yields breakpoint values for 10% ethanol (10E) that are stable and comparable to those measured for other drugs of abuse [Alcohol. Clin. Exp. Res. 23 (1999) 1580]. The aims of the present experiment were twofold: (1). to determine whether this procedure is sensitive to changes in reinforcer magnitude using a reinforcer previously demonstrated to affect operant responding in a predictable fashion and (2). to determine whether ethanol reinforcement produced similar changes in behavior. Male, Long-Evans rats were trained to respond for either 3% sucrose (3S) or 10E using the sipper tube appetitive/consummatory procedure where the completion of a single response requirement results in access to a liquid solution for 20 min. Three successive breakpoints were determined for this "baseline" solution by increasing the response requirement each day until it was not completed. The concentration of the solutions was then manipulated such that breakpoints for the Sucrose Group were assessed for 1%, 3%, 5% and 10% sucrose, and breakpoints for the Ethanol Group were assessed for 2%, 5%, 10% and 20% ethanol. The concentration manipulation showed that sucrose concentration had a greater impact on seeking and consumption than did ethanol concentration. Breakpoints in the Sucrose Group were highly correlated with sucrose concentration, whereas in the Ethanol Group, breakpoint was unrelated to ethanol concentration. Ethanol intake patterns suggested that pharmacological factors might have been regulating intake, and that when physiologically detectable amounts of ethanol were consumed, there was a dissociation between seeking and intake with slightly elevated ethanol seeking. Overall, the across-session breakpoint procedure confirmed that sweet taste was highly related to seeking and consumption, whereas ethanol-motivated responding may be controlled by different regulatory mechanisms that are distinct to seeking and consumption. PMID- 12536011 TI - Hormonal control of grooming behavior in domestic goats. AB - In this study, further documentation of the modulation of self-grooming by testosterone was provided by an experiment which showed that self oral grooming in long-term castrated male goats of the dwarf Shiba breed was significantly down regulated to one-third of the baseline grooming rate by testosterone supplementation. Observations on grooming in gonadally intact female Shiba goats in the summer (July) and in the fall (October) showed that the oral grooming rate was significantly higher in the fall, occurring at about twice the rate in the summer. The female oral grooming rate in the fall was the same as the baseline rate of castrated males before testosterone supplementation. The seasonal increase in females was not attributed to changes in gonadal hormones. The adaptive significance of hormonal regulation of the timing mechanism for grooming in both male and female goats, and possibly other ungulates, is discussed. PMID- 12536012 TI - Effect of naltrexone on food intake and body weight in Syrian hamsters depends on metabolic status. AB - Opioids are a family of neuropeptides involved in the control of food intake and regulation of body weight. In general, nonselective opioid antagonists have inhibited food intake in a variety of paradigms in rodent species. Syrian hamsters may be an exception to the general findings. In a previous report, we showed that systemic administration of an opioid antagonist, naltrexone, for 2 days increased body weight in female Syrian hamsters. To confirm the extent of these finding we designed the present experiment testing the effect of a chronic 6-day infusion of naltrexone on food intake, water intake, and body weight in freely feeding male hamsters. In addition, we examined the effect of acute administration of naltrexone on food intake in both ad-libitum-fed and food deprived hamsters. We found that chronic systemic administration of naltrexone caused a significant increase in food intake and body weight. Second, acute administration of naltrexone decreased food intake after a 48-h fast but had no effect in ad-libitum-fed hamsters. Water consumption was not altered in any experimental paradigm. Our results suggest that opioid circuits in Syrian hamsters may function tonically to suppress food intake and body weight when Syrian hamsters are in positive energy balance. Paradoxically, opioids may enhance food intake after a sustained fast. PMID- 12536013 TI - Effects of tGLP-1 on feeding behaviour and neurohypophysial function under chronic osmotic stimulation. AB - Effects of repeated daily intracerebroventricular injections of 1 microg of glucagon-like peptide-1 (7-36) amide (tGLP-1) on feeding and drinking behaviour, as well as neurohypophysial hormone secretion, was investigated in rats drinking tap water or 2% saline for 6 days. In euhydrated rats, tGLP-1 decreased moderately food and water consumption without a marked reduction of body weight. In salt-loaded rats, tGLP-1 considerably inhibited saline intake. On the other hand, food consumption and body weight changes were similar in vehicle- and tGLP 1-treated rats drinking 2% saline. Osmotic stimulation resulted in the augmented release of both neurohypophysial hormones. tGLP-1 did not alter plasma vasopressin and oxytocin concentrations either in euhydrated or osmotically stimulated rats. It is concluded that tGLP-1 may modify feeding and drinking behaviour under conditions of normal or disturbed water-electrolyte balance in the rat. PMID- 12536014 TI - Lambs prevented from suckling their mothers display behavioral, immune and endocrine disturbances. AB - The aim of the experiment was to assess the effects of either maternal deprivation or prevented suckling on lambs. Thirty Comisana lambs were assigned to a control dam-suckled group (DS) and two test groups of 10 each. Test lambs were maintained with their mothers 24 to 30 h after parturition and subsequently offered ewe milk from buckets: EM lambs were separated from their mothers, while EM+D animals were reared with their dams but suckling was denied by covering ewe udders with juta bags. Behavioral, immune and cortisol responses of lambs were assessed throughout the experiment. EM+D lambs spent less time (P<.001) on investigative behavior compared to the two other groups. When isolated in a novel environment, EM+D lambs exhibited a shorter duration of movement (P<.01) and a longer latency time compared to DS and EM lambs (P<.001 and P<.05, respectively) as well as a higher number of bleats (P<.01) than EM lambs. EM+D lambs also displayed higher (P<.001) plasma cortisol levels than the other two groups when isolated at 4 days of age. When subjected to discrimination tests, EM+D lambs spent less time near their companions (P<.01) than EM lambs and took a longer time to reach their pen mates (P<.01) than DS and EM lambs. EM+D lambs displayed reduced growth (P<.001) compared to DS animals during the 0-7 and 8-14-day periods. We conclude that frustration arising from maternal feeding deprivation results in altered endocrine and behavioral responses and reduced growth suggesting emotional disturbances of lambs. PMID- 12536015 TI - Strain differences in hippocampal EEG are related to strain differences in behaviour in rats. AB - To date, EEG studies towards strain differences have focussed on pharmacologically altered or pathological EEG activity, but only few studies have investigated strain differences and normal EEG activity. A strong relation between behaviour and EEG activity has been demonstrated, especially for hippocampal EEG activity. This relation is known to be similar across species and strains, but no direct comparisons between rat strains within one study have been made. This study compared two rat strains (Sprague-Dawley and Long-Evans) with regard to open-field behaviour and concurrent hippocampal EEG recordings. A detailed behavioural analysis was made and spectral power was calculated for corresponding EEG activity in eight frequency bands. The two strains differed in exploratory activity and in spectral power in the 9-10-Hz frequency band (high frequency rhythmical slow activity [RSA] 6-10 Hz). Long-Evans rats showed higher exploratory activity and higher 9-10 Hz spectral power for voluntary movement and sniffing behaviours. Our results demonstrated these behaviour-specific strain differences in RSA power, although the relation between EEG and behaviour within each strain was similar. The strain differences in EEG were interpreted in relation to strain differences in exploratory behaviour, attributing the differences to a main motor component but also to a smaller sensory component integrated in exploratory behaviour. This is in accordance with theories on the sensory-motor function of the hippocampus and hippocampal theta activity. PMID- 12536016 TI - Combined effects of temperature and noise on human discomfort. AB - The trade-off between noise and temperature and their combined effects on discomfort were studied on 108 lightly clothed subjects (0.6 clo), individually exposed for 2 h in a climatic chamber. Every 10 min of the first hour, subjects could modify the experimental conditions by deciding a change in temperature or noise. However, any change in one parameter was experimentally associated with a fixed change in the other parameter according to eight predetermined designs and all trials for thermal improvement were detrimental to acoustic comfort and conversely. Four initial exposures started at thermoneutrality (24 degrees C) in a noisy environment (85 dBA, recorded fan noise), the reduction of noise being linked to a temperature change towards cool or warm climates. The other four conditions started at a low noise level (35 dBA) but in a cool (14 or 19 degrees C) or warm (29 or 34 degrees C) environment, the reduction of thermal discomfort towards 24 degrees C leading to a louder noise. After six possible voluntary changes, the environment was kept constant for 1 h. Ambient parameters, skin temperatures, and subjective estimates were recorded. Results showed that females accepted noisier environments than males, suggesting that thermal comfort is dominant for women. Noise was rated as the most unpleasant factor when initial conditions were noisy whereas temperature was the most disturbing factor when subjects began the experiment with thermal conditions far from thermoneutrality. Finally, although the combined effects of noise and temperature did not influence the physiological data, our results suggest that noise may alter thermal pleasantness in warm conditions. PMID- 12536017 TI - Vigorous intake of oil emulsion caused by chronic food deprivation remains after recovery in rats. AB - Oil emulsion intake over a 30-min period was compared under different dietary conditions (ad libitum feeding and chronic food deprivation) and at various concentrations of oil in rats. The pattern of intake for each dietary condition was extremely different. Food-deprived rats ingested more emulsion when the solution was thicker but intake amount became less at too high a concentration. Ad libitum feeding rats ingested less emulsion than deprived rats with no difference among the concentrations. Rats on a restricted diet clearly differentiated between concentrations, selecting the thicker emulsion when provided two-bottle selection between 10% oil and some other concentration. On the other hand, rats fed ad libitum differentiated between only extremely weak solutions. The vigorous intake of oil emulsion induced by chronic food deprivation was maintained after 2 weeks of normal feeding. Significant difference between prior dietary conditions was maintained at lower concentrations. Dietary timing for food-deprived rats affected little on emulsion intake. These results indicate that response to oil emulsion intake differs by concentration in rats. PMID- 12536018 TI - Air and shock two-way shuttlebox avoidance in C57BL/6J and 129X1/SvJ mice. AB - Despite multiple advantages of the use of electric shock as an aversive stimulus, reasons exist for considering alternative aversive stimuli. In the present study, we examined and compared the acquisition of two-way shuttlebox avoidance with 275.8-kPa (40-psi) pulsed air and continuous 0.4-mA shock in two strains of mice commonly employed in targeted gene mutation research, C57BL/6J and 129X1/SvJ. Each trial consisted of a 5-s warning stimulus (WS, light) during which shuttling to the other side cancelled delivery of the aversive stimulus. Once initiated, the aversive stimulus remained active for 20 s or until an escape response occurred. For C57BL/6J mice, air and shock were equally and highly effective aversive stimuli. In contrast, air was less effective than shock for 129X1/SvJ mice. C57BL/6J mice outperformed 129X1/SvJ mice for both stimulus types. For 129X1/SvJ mice, longer escape latencies were observed initially for air, suggesting that shock is more effective. However, these differences in latency dissipated within the first seven sessions. Nevertheless, by the end of the 17 day study, asymptotic levels of avoidance proficiency were substantially lower for air than for shock in 129X1/SvJ mice. These results indicate that air is a suitable substitute for shock as an aversive stimulus in shuttlebox active avoidance; however, the relative efficacies of these aversive stimuli appear to depend upon the strain chosen for study. PMID- 12536019 TI - Longitudinal study of daily variation of rats' behavior in the elevated plus maze. AB - We conducted a longitudinal study about daily variation of Wistar male rats' behavior in the elevated plus-maze (EPM) evaluated in the 1st, 2nd, 3rd, 6th, 12th, and 18th months of life. Animals were submitted to the plus-maze in 12 sessions at 2-h intervals (n=72, 6 per time point). Spontaneous rest-activity rhythm of four animals was assessed by observation of 24-h videotape records. Time series were analyzed by Cosinor method. Behavioral rates on the six occasions and in light and dark phases were compared by means of two-way ANOVA with repeated measures. Exploratory behavior in EPM was smaller in the light phase and in older animals. Higher values of open and closed arms exploration were observed in the first and third months of the dark phase, and in the first month of the light phase. Adjustment to the 24-h period was significant at all stages for rest-activity data, number of entries in closed arms, and time on center, and for three to five stages for open-arm exploration. In general, 24 h variability was more pronounced in younger animals compared with older ones. The present study showed that: (1). a significant amount of total variability of the behavioral indexes analyzed could be attributed to 24 h variation, (2). light/dark phases differences in EPM exploration were present at all developmental stages, (3). older Wistar rats explored less the EPM and were less active in their home cage compared with younger ones, and (4). behavioral indexes (EPM) decrease was phase related and partially related to a reorganization of rest-activity rhythm. PMID- 12536020 TI - Consolidation and expression of a shock-induced odor preference in rat pups is facilitated by opioids. AB - To support nipple attachment and huddling, rat pups must learn to approach and prefer maternal odor. Similar to other altricial species, rat pups have a sensitive period for learning this odor preference, which ends around postnatal day (PN) 10 and coincides with the emergence of walking. One characteristic of this sensitive period is that an odor paired with moderate shock elicits an odor preference. After PN10, this behavioral training produces an odor aversion, although pain threshold remains unchanged. Recently, we demonstrated that the endogenous opioid system might be a key element in the acquisition of the shock induced odor preference during the sensitive period since antagonism of this system disrupts odor preference learning. In older pups, acquisition of a shock induced odor aversion was unaffected by opioid system manipulation. The purpose of these experiments was to further elucidate the role of opioids in infant olfactory learning through assessment of memory consolidation and expression during and after the sensitive period. In Experiment 1, we demonstrate that naltrexone (NTX), a nonspecific opioid antagonist, given immediately following odor-shock conditioning during the sensitive period, blocks odor preference formation and yields an odor aversion. However, the same treatment does not disrupt consolidation of an odor aversion in older pups. In Experiment 2, we demonstrate that during the sensitive period, NTX disrupts expression of the shock-induced odor preference, but not the learned odor aversion in older pups. Results using this model of attachment suggest that opioids have an important role in the acquisition, consolidation, and expression of early olfactory preferences. Furthermore, since prenatal drug exposure is known to alter the endogenous opioid system, these results highlight the capacity of prenatal opiate exposure to disrupt early infant learning and attachment. PMID- 12536021 TI - Meal size, satiety and cholecystokinin in gastrectomized humans. AB - A wealth of data supports the idea that the stomach and cholecystokinin octapeptide (CCK-8) normally play important roles in meal size and satiety. We studied long-term gastrectomized humans to further evaluate this possibility. Ten humans, who were gastrectomized 8 (3-12) years earlier, and eight controls ate a meal from a plate placed on a scale connected to a computer and estimated their satiety every minute using a computerized rating scale. Blood levels of CCK-8 were measured before and after the meal. There was no difference between the groups in the amount of food consumed or in the perception of satiety during the meal. Gastrectomized humans had higher blood levels of CCK-8 than controls before the meal; the levels increased after the meal in the controls but not in the gastrectomized subjects. It is suggested that although the stomach and CCK-8 normally are involved in the control of meal size and satiety, their roles are dispensable. PMID- 12536022 TI - Expression of biologically active rat apolipoprotein AIV in Escherichia coli. AB - Rat apolipoprotein AIV (apo AIV) is a 43-kDa intestinal apolipoprotein that is important in lipid metabolism and the suppression of food intake. In this study, a full-length rat apo AIV was expressed in Escherichia coli and purified in a bioactive form. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) and mass spectrometric analysis revealed that the isolated recombinant protein has a molecular mass of approximately 43 kDa, similar to that of natural rat apo AIV. Immunoblot analysis and N-terminal amino acid sequencing confirmed the identity of the recombinant apo AIV protein as natural rat apo AIV. The recombinant protein was functional in lipoprotein binding assays. Biological activity was assessed behaviorally in that the recombinant protein suppressed food intake of fasted rats comparably to natural rat apo AIV. Neither native nor recombinant apo AIV elicited a conditioned taste aversion (CTA) at doses that suppress feeding. These results indicate that the recombinant apo AIV is structurally and functionally indistinguishable from rat natural apo AIV, making this overexpression and purification scheme a powerful tool for future structure and function studies. PMID- 12536023 TI - Effects of N-methyl-D-aspartate-induced amygdala lesion in rats submitted to the elevated T-maze test of anxiety. AB - The elevated T-maze (ETM) has been used to generate two defensive behaviors in the same rat, inhibitory avoidance and one-way escape, which have been related to generalized anxiety and panic, respectively. In the present study, we investigate the role of the amygdala on the modulation of these two behaviors. Male Wistar rats were tested in the ETM test 2, 7, or 14 days after bilateral N-methyl-D aspartate (NMDA)-induced lesion of the amygdala. The animals were also tested in an open field for evaluation of motor performance. The results showed that animals tested 7 days after NMDA injection had impairment in the acquisition of inhibitory avoidance, indicating an anxiolytic effect. Lesion of the amygdala did not change one-way escape in any of the tested groups. These results provide further evidence for the involvement of the amygdala in the modulation of defensive behaviors that have been associated to generalized anxiety. PMID- 12536024 TI - The relation between saliva flow after different stimulations and the perception of flavor and texture attributes in custard desserts. AB - Salivary flow rates were measured at rest and after three types of stimulation; odor, Parafilm chewing, and citric acid. The highest flow rate was elicited by citric acid followed by Parafilm and odor, while the lowest flow rate was unstimulated. In order to investigate whether and how the amount of saliva a subject produces influences the sensory ratings, the four types of salivary flow rates were correlated with sensory ratings of three different types of vanilla custard dessert. No significant correlation could be found between any of the salivary flow rates and the sensory ratings. A subject with a larger saliva flow rate during eating did not rate the foods differently from a subject with less saliva flow. The same pattern was seen for all types of stimulation. This finding could indicate that subjects are used to their respective amounts of saliva to such a degree that the differences in sensory ratings between subjects cannot be explained by the interindividual difference in saliva flow rate. PMID- 12536026 TI - Iron deficiency and the intellect. AB - Children are especially liable to iron-deficiency anaemia in developing countries, and in the inner cities of developed countries. Does the lack of iron cause impaired physical and mental development, and can this in certain circumstances be a permanent effect? One of the reasons that this is such a difficult question to answer is that there can be so many confounding factors, from other nutritional deficiencies, to helminthic infections and malaria in tropical countries. If there is a definite relationship, children in the first 2 years of life will be at particular risk during the mayor spurt of brain growth. Lack of iron can affect brain cells, myelin, or neurotransmitters, so there is certainly a theoretical basis for possible brain damage, or there could be an effect from lack of oxygen. Also anaemic children are likely to feel ill and unwilling to co-operate with tests to assess for developmental defects. Many studies of the possible results of iron deficiency on the development of children have been carried out in various countries, and some of these from 1983 onwards are recorded. It is difficult to draw conclusions from these trials, partly due to the variability in their construction, but on balance the evidence suggests that treatment of iron deficiency is justifiable, whether this is associated with anaemia or not. It is equally important to stress the importance of prevention, although more research is needed on the best method to use, which is both effective and affordable. PMID- 12536027 TI - Virological and immunological aspects of seizure disorders. AB - The brain is a symptom-producing organ, and one of the symptoms due to a basic brain dysfunction is epilepsy. The pathophysiologic background is in most epilepsies multifactorial, as different pre-, peri-, and postnatal triggers or environmental conditions influence one or several genetic factors, where also gender is of importance. One of the genetic factors is immunodysfunction, and the trigger mechanism may be a virus infection. Viruses are the most common agents to which the human being is exposed throughout life. The herpes virus group is of special interest with respect to complications of the central nervous system. Herpes viruses, especially herpes simplex virus type 1 (HSV-1), human herpes virus type 6 (HHV-6), cytomegalovirus (CMV) and Epstein-Barr virus (EBV), are capable of establishing latent infection and reactivating under a variety of stimuli. In this review especially HHV-6 will be emphasized, as well as CMV in relation to Rasmussen's syndrome. The immunological aspects will focus on immunoglobulins, antibodies, especially the glutamate receptors, human leukocyte antigens, T- and B-lymphocytes, and their respective interaction with the antigen presenting cell. This course of events concerns the 'immunological synapse'. Finally, reports on herpes virus genomes in the human brain are discussed. A study on herpes viral DNA in brain tissue from patients operated for focal epilepsy is briefly mentioned. PMID- 12536028 TI - Characteristics of dipoles in clustered individual spikes and averaged spikes. AB - The aim of this study is to analyze the characteristics of dipoles in clustered individual spikes and averaged spikes, we compared electroencephalography (EEG) dipole localizations from patients with intractable extratemporal lobe epilepsy (IETLE) and from patients with benign epilepsy with centrotemporal spikes (BECTS). We studied 10 patients; five with IETLE who underwent epilepsy surgery after subdural EEG and five with BECTS. We recorded 19-channel digital scalp EEGs and used clustering analysis for individual spikes to characterize interictal spikes. We selected and averaged one representative spike group at the maximum negative peak electrode. We used a single dipole method with three-shell spherical head model. We compared dipole localizations of both averaged and individual spikes.IETLE data had more identifiable spike clusters and fewer spikes in each cluster than BECTS (P<0.05). Dipole sources with goodness-of-fit >or=95% in averaged spikes were less frequent in IETLE than in BECTS (P<0.05). For IETLE, averaged spikes showed no dipoles (two patients), while individual spikes gave dipole sources reliably in the epileptic region. For BECTS, individual and averaged spike sources were clustered. More than 80% of dipoles in averaged spikes were stable, in close proximity, for prolonged periods in BECTS. More spike groups after clustering and fewer acceptable dipoles from averaged spikes in IETLE reflect variable spike activity over extensive epileptic regions. Fewer spike groups producing more acceptable dipoles in BECTS correlate with stable spike sources within the isolated epileptic central region. Characteristics of clustered interictal spikes need careful examination before the use of dipole analysis of averaged spikes for epilepsy evaluation. PMID- 12536029 TI - Muscle ultrasonography and electromyography correlation for evaluation of floppy infants. AB - Floppiness in an infant may have a number of different etiologies from disorders of the brain to spinal cord lesions, neuropathies, neuromuscular junction disorders and myopathies. In this study we aimed to investigate the correlation of muscle ultrasonography (US) and electromyography (EMG) in the diagnosis of floppy infants. The study encompassed 41 floppy infants aged 2-24 months. The muscle US and EMG examinations were performed without awareness of the clinical diagnosis. The final diagnosis was established by molecular genetic tests or muscle/nerve biopsy. The neurogenic group consisted of 16 infants according to their US and EMG findings. Fifteen of them had spinal muscular atrophy proven by genetic analysis and one had polyneuropathy diagnosed by nerve biopsy. Six infants were in the myopathic group according to their muscle US and EMG results. All of them underwent muscle biopsy and microscopic examination revealed five congenital muscular dystrophy and one glycogen storage disease. In two infants the US and EMG data conflicted. Their biopsies were also insufficient for the diagnosis. Seventeen infants had normal US and EMG findings but pathologic cranial magnetic resonance imaging or metabolic/genetic tests. They were considered in the group of central hypotonia. Our results suggest a high concordance of US and EMG findings in the diagnostic work-up of neurogenic and myopathic disorders. PMID- 12536030 TI - Apnea associated with hypoxia in preterm infants: impact on cerebral blood volume. AB - The present study analyzed changes in cerebral blood volume (CBV) during apnea associated with hypoxia compared to apnea without hypoxia. Hypoxia was defined as pulsoxymetric oxygen saturation <80%>10 s. The employed technique was near infrared spectroscopy combined with electrocardiogram, electroocologram, pulsoxymetry, sidestream capnography and two respiratory effort sensors. In 24 preterm infants 44 incidences of apnea were analyzed. Two main patterns were observed: a significant decrease or a significant increase of CBV. In the 'CBV decrease' group deltaCBV was -55 microl/100 g brain in hypoxic apnea, and -62 microl/100 g brain in non-hypoxic apnea. In the 'CBV increase' group the rise of CBV above preapneic values was +50 microl/100 g brain in hypoxic apnea, and +47 microl/100 g brain in non-hypoxic apnea. Heart rate showed a significant decrease only in the 'CBV decrease' group. Endexpiratory CO(2) increased significantly 1 min after apnea. In conclusion, this study observed significant changes of CBV during apnea in preterm infants, but no difference in CBV behavior regarding whether incidents of apnea were associated with hypoxia or not. It remains unclear which regulatory mechanisms are responsible for the two observed patterns of deltaCBV during apnea. PMID- 12536031 TI - Dendritic overgrowth and alterations in laminar phenotypes of neocortical neurons in the newborn with semilobar holoprosencephaly. AB - We analyzed neuronal phenotypes and dendritic growth in the newborn with semilobar holoprosencephaly and 18p deletion. We found that the holoprosencephalic neocortex retained its basic six-layered lamination but displayed a number of intralaminar and modular architectonic alterations and contained a mixture of normal and aberrant neuronal phenotypes. The most conspicious finding was the presence of the pronounced increase in soma size and total basal dendritic length of holoprosencephalic layer III pyramidal neurons in comparison to age-matched control brains. The dramatic (5-fold) dendritic overgrowth observed in associative cortico-cortical pyramidal neurons is probably related to the pronounced diminution of the cortical afferent input. PMID- 12536032 TI - Magnetic resonance imaging in late-onset ornithine transcarbamylase deficiency. AB - We examined brain magnetic resonance imaging (MRI) in a cohort of seven patients with ornithine transcarbamylase deficiency (OTCD), and correlated MRI findings with clinical manifestations. Seven patients with OTCD, aged 3-27 years, all with a missense mutation, were involved in the study. We classified the OTCD patients clinically into four stages. MR study was performed with a 1.5-T superconducting magnet during asymptomatic periods. MRI revealed white matter lesions in two patients with an advanced clinical stage, i.e. T1 and T2 prolongated round lesions in the deep white matter and posterolateral angle of the lateral ventricle in one patient; small foci of T2 and T1 prolongation in the subcortical white matter in another. Parenchymal lesions, and cerebral and cerebellar atrophy were not found in the other five patients. MRI might be normal in the early stage of the disease, and progress in proportion to the clinical stage of OTCD. OTCD should be considered as a differential diagnosis of small foci in the white matter in children. PMID- 12536033 TI - Cerebral proton magnetic resonance spectroscopy of a patient with giant axonal neuropathy. AB - Magnetic resonance imaging of a girl with giant axonal neuropathy revealed a progressive white matter disease. In close agreement with histopathological features reported previously, localized proton magnetic resonance spectroscopy at 9 and 12 years of age indicated a specific damage or loss of axons (reduced N acetylaspartate and N-acetylaspartylglutamate) accompanied by acute demyelination (elevated choline-containing compounds, myo-inositol, and lactate) in white matter as well as a generalized proliferation of glial cells (elevated choline containing compounds and myo-inositol) in both gray and white matter. PMID- 12536034 TI - Anti-epileptic drugs-induced de novo absence seizures. AB - The authors present three patients with de novo absence epilepsy after administration of carbamazepine and vigabatrin. Despite the underlying diseases, the prognosis for drug-induced de novo absence seizure is good because it subsides rapidly after discontinuing the use of the offending drugs. The gamma aminobutyric acid-transmitted thalamocortical circuitry accounts for a major part of the underlying neurophysiology of the absence epilepsy. Because drug-induced de novo absence seizure is rare, pro-absence drugs can only be considered a promoting factor. The underlying epileptogenecity of the patients or the synergistic effects of the accompanying drugs is required to trigger the de novo absence seizure. The possibility of drug-induced aggravation should be considered whenever an unexpected increase in seizure frequency and/or new seizure types appear following a change in drug treatment. By understanding the underlying mechanism of absence epilepsy, we can avoid the inappropriate use of anticonvulsants in children with epilepsy and prevent drug-induced absence seizures. PMID- 12536035 TI - Linear scleroderma associated with progressive brain atrophy. AB - Linear scleroderma (LS) is characterized by scleroatrophic lesions affecting limbs and legs, unilaterally. Neurological involvement may be associated with ipsilateral facial and skull involvement in disorders referred to clinically as LS 'en coup de sabre', and Parry-Romberg syndrome. We report a child with LS presenting with a severe neurological disorder characterized by epilepsy, progressive mental deterioration and a rapid process of atrophy involving the ipsilateral cerebral hemisphere, but not associated with an overlying facial structure involvement. Functional brain studies showed a reduction in the diameter of the left internal carotid and of the left middle cerebral artery. Our observations suggest that neuroimaging studies should be considered in all patients with linear scleroderma, and such studies become necessary when neurological symptoms occur. PMID- 12536036 TI - Centronuclear myopathy and cardiomyopathy requiring heart transplant. AB - Centronuclear myopathy has been extremely rarely associated with cardiomyopathy, which can lead to heart failure and premature death. We report the case of a 3.5 year-old girl with early-onset dilated cardiomyopathy, biventricular hypertrophy and histologic features suggestive of centronuclear myopathy. After unsuccessful medical treatment for heart failure, she underwent cardiac transplantation at the age of 4.5 years. Results of a skeletal muscle biopsy showed increased central nuclei and perinuclear vacuolations with aggregates of mitochondria. Examination of the heart at the time of transplantation confirmed a diagnosis of dilated cardiomyopathy. Histologic results revealed hypertrophic myocardiocytes, focal areas of infarction and endocardial fibroelastosis, most prominently in the left ventricle. Although cardiomyopathy is commonly associated with other childhood myopathies, to our knowledge, this is the youngest patient reported with centronuclear myopathy presenting with heart failure caused by cardiomyopathy, and the first patient to successfully undergo cardiac transplantation. One year after the heart transplant, there were no signs of rejection. We recommend detailed cardiac assessment with regular follow-up for children with histologic features consistent with centronuclear myopathy. PMID- 12536037 TI - The 6th Scientific Meeting of the Taiwan Child Neurology Society, Taipei, 18 May 2002. PMID- 12536038 TI - Serotonin receptor mRNA expression in rat dorsal root ganglion neurons. AB - In the present study, we have used in situ hybridization to examine the distribution of serotonin (5-HT) receptors in rat dorsal root ganglion (DRG) neurons. Within DRG neurons, mRNAs for 5-HT1B, 5-HT1D, 5-HT2A, 5-HT2B, 5-HT3B and 5-HT4 receptors were readily detected in small (<25 microm), medium (25-45 microm) and large (>45 microm) diameter neurons. In contrast mRNAs for 5-HT1A, 5 HT1E, 5-HT2C, 5-HT5A, 5-HT5B, 5-HT6 and 5-HT7 receptors were undetectable in these neurons. The present study provides an insight into the molecular profile of 5-HT receptor subtypes in neurons responsible for modulating sensory information. PMID- 12536039 TI - Short-term effects of 3,4-methylenedioximethamphetamine on noradrenergic activity in locus coeruleus and hippocampus of the rat. AB - This study was undertaken to investigate the effects of the administration of 3,4 methylenedioxymethamphetamine (MDMA) on the locus coeruleus firing rate, on the sensitivity of the alpha(2)-adrenoceptors which regulate neuronal activity and on the in vivo tyrosine hydroxylase activity in hippocampus. The basal firing rate was not modified by either a single dose or repeated doses of MDMA, although the latter produced a shift to the right in the dose-response curve for clonidine induced inhibition of the firing rate (ED(50) increased by 59%) and a reduction in tyrosine hydroxylase activity (20%) in the hippocampus. However, 8 days after the final dose alpha(2)-adrenoceptor sensitivity and tyrosine hydroxylase activity had returned to control values. Our results show a desensitization of alpha(2)-adrenoceptors in locus coeruleus and the existence of short-term changes in the noradrenergic system. PMID- 12536040 TI - Two classes of astrocytes in the adult human and pig retina in terms of their expression of high affinity NGF receptor (TrkA). AB - Astrocytes have been implicated in axon guidance and synaptic regeneration in the retina and these processes involve activation of the high affinity nerve growth factor receptor, known as the tyrosine kinase A (TrkA) receptor. The purpose of the present study was to characterize the expression of TrkA in astrocytes of the adult pig and human retina. To this end, sections of human and pig retinas were immunolabeled with a combination of antibodies to glial fibrillary acidic protein (GFAP) and TrkA. Our study revealed that most of the GFAP-positive cells express TrkA, whereas a rare, novel subpopulation of astrocytes was found to be devoid of TrkA. Our results support the idea that astrocytes play an important neurotrophic role in the retina. PMID- 12536041 TI - Central administration of cocaine- and amphetamine-regulated transcript inhibits food intake in chicks. AB - The present study was done to clarify whether intracerebroventricular (ICV) injection of cocaine- and amphetamine-regulated transcript (CART) affects feeding in chicks. ICV injection of CART significantly inhibited fasting-induced feeding of broiler chicks. In layer chicks, on the other hand, CART inhibited food intake in birds with ad libitum access to feed but only weakly affected intake of fasted birds. In addition, the ICV injection of CART attenuated neuropeptide Y (NPY) induced feeding in both broiler and layer chicks. These results indicate that CART is one of the important regulators of feeding in chicks, but the suppressive effect on feeding is somewhat different between strains. Furthermore, the present study also demonstrates that CART interacts with NPY in the central nervous system to regulate feeding in chicks. PMID- 12536042 TI - Nitric oxide inhibits adenosine 5'-triphosphate-induced Ca2+ response in inner hair cells of the guinea pig cochlea. AB - To investigate the interaction between Ca(2+) and nitric oxide (NO) in inner hair cells of the guinea pig cochlea (IHCs), the extracellular adenosine 5' triphosphate (ATP)-induced NO production and the effects of NO on ATP-induced increase of intracellular Ca(2+) concentrations ([Ca(2+)](i)) were investigated in IHCs using the NO-sensitive dye DAF-2 and the Ca(2+)-sensitive dye Fura-2. Extracellular ATP induced an increase in DAF-2 fluorescence, which thus indicates NO production in IHCs. The ATP-induced NO production was mainly due to Ca(2+) influx through the activation of P2 receptor. L-N(G)-nitroarginine methyl ester, a NO synthesis inhibitor, enhanced the ATP-induced [Ca(2+)](i) increase in IHCs while S-nitroso-N-acetylpenicillamine, a NO donor, inhibited it. We conclude that NO inhibits the ATP-induced [Ca(2+)](i) increase in IHCs by a negative-feedback mechanism. PMID- 12536043 TI - Facilitatory action of halothane at subanesthetic concentrations on glutamatergic excitatory synaptic transmission in the CA1 area of adult rat hippocampus. AB - Whole-cell recordings were made from pyramidal cells visually identified in the CA1 field of adult rat hippocampal slices, and the effects of subanesthetic concentrations of halothane on excitatory postsynaptic currents mediated by non-N methyl-D-aspartate subtype glutamate receptors were investigated. Halothane concentrations were measured by gas chromatography. At concentrations of 0.2 mM and 0.6 mM, halothane reversibly decreased the amplitude of excitatory postsynaptic currents (EPSCs) evoked by electrical stimulation of Schaffer collateral fibers, and the decrease was accompanied by enhanced paired-pulse facilitation, consistent with the previously reported presynaptic site of halothane's inhibitory action. By contrast, at lower concentrations (0.02 mM and 0.05 mM), halothane increased the amplitude of EPSCs without any appreciable changes in paired-pulse facilitation. Moreover, the frequency of miniature EPSCs arising spontaneously in the presence of tetrodotoxin (mEPSCs) was increased by subanesthetic halothane, but the amplitude of the mEPSCs did not change significantly. These observations suggest that at subanesthetic concentrations halothane postsynaptically enhances glutamatergic excitatory synaptic transmission. This may provide a vital clue to elucidation of the neural mechanisms of the nociceptive reflex enhancement and excitatory state that occur at light levels of anesthesia. PMID- 12536044 TI - Somatostatin receptor immunoreactivity in the eye of the adult newt (Pleurodeles waltlii Michan). AB - The neuropeptide somatostatin is found in the retina of many species, yet its role in the visual process remains to be elucidated. The aim of the present study was to examine the expression and cellular localization of somatostatin receptor subtypes (sst; sst(2A), sst(2B) and sst(3)) in the eye of the adult newt Pleurodeles waltlii using immunohistochemistry. sst(2A) immunoreactivity was observed in bipolar cells, in the inner segments of cone photoreceptors, as well as in the region corresponding to connecting cilia of rods. sst(2B) immunoreactivity was not detected. sst(3) immunostaining was localized most intensely in the inner segments of cones, and in cilia of rods. These results suggest that somatostatin acting via sst(2A) and sst(3) receptors may play an important role in retinal physiology of the lower vertebrates. PMID- 12536045 TI - A comparison of hyperalgesia and neurogenic inflammation induced by melittin and capsaicin in humans. AB - Melittin (a main compound of bee venom) and capsaicin were injected intradermally in healthy human volunteers: (1) to study secondary mechanical hyperalgesia (static hyperalgesia and dynamic hyperalgesia) around the injection site; and (2) to correlate the sensory changes to the neurogenic inflammation assessed by laser doppler blood flowmetry. Melittin 50 microg and capsaicin 10 microg induced comparable spontaneous pain and increased blood flow (neurogenic inflammation). Intradermal injection of melittin induced regions of secondary mechanical hyperalgesia around the injection site, however, they were not as large as the hyperalgesia induced by capsaicin. This is the first report studying mechanical hyperalgesia induced by melittin in humans, and the results were in agreement with the previous observations in rats. Melittin seems to be a valuable model to study a possible contribution of neurogenic inflammation to hyperalgesia in humans. PMID- 12536046 TI - Investigation of the complex descending innervation of the dorsal cochlear nucleus in the rat: a transneuronal tract-tracing study using pseudorabies virus. AB - The afferent neuronal connections of the dorsal cochlear nucleus were investigated in rats by using a trans-synaptic retrograde tract-tracing method. The neurotropic viral tracer, the Bartha strain of the pseudorabies virus was stereotaxically injected into the dorsal cochlear nucleus, ipsilaterally. Neurons, which project directly or indirectly (one or multiple relays by other neurons) to the dorsal cochlear nucleus were infected and visualized by immunohistochemistry. Labeled neurons were found in each components of the auditory pathway, some of the monoaminergic cell groups in the lower brainstem, the hypothalamus and in some limbic areas. PMID- 12536047 TI - Acupuncture decreases nitric oxide synthase expression in periaqueductal gray area of rats with streptozotocin-induced diabetes. AB - Acupuncture has been used as a clinical treatment in Oriental medicine for various diseases including diabetes mellitus, one of the most common metabolic disorders in humans. In the present study, the effect of acupuncture on the expressions of neuronal nitric oxide synthase (nNOS) and nitric oxide synthase (NOS) in the dorsolateral periaqueductal gray (DL-PAG) area of rats with streptozotocin (STZ)-induced diabetes was investigated via nNOS immunohistochemistry and nicotinamide adenine dinucleotide phosphate-diaphorase histochemistry. Enhanced expression of nNOS and NOS was detected in the DL-PAG of rats with STZ-induced diabetes, and acupunctural treatment at Zusanli acupoint suppressed the diabetes-induced enhancement in the expression of nNOS and NOS. The present results demonstrate that acupuncture is effective in the modulation of the expression of nNOS and NOS in the DL-PAG under diabetic conditions. PMID- 12536048 TI - Synaptic mitochondrial changes in the motor cortex following unilateral cortical lesions and motor skills training in adult male rats. AB - Unilateral lesions of the forelimb sensorimotor cortex and motor skills training interact to enhance synaptic plasticity in layer V of the motor cortex contralateral to the lesion in male rats. In this study, we assessed the metabolic correlates of these synaptic changes by quantifying the number and size of mitochondria in synaptic axonal terminals with perforated or non-perforated post-synaptic densities (PSDs) and single or multiple post-synaptic contacts. The number of synaptic mitochondria per neuron was significantly greater in rats with lesions. Neither manipulation affected mitochondrial size or number of mitochondria per synapse. Independent of experimental condition, terminals with perforated PSDs had more mitochondria than those with non-perforated PSDs and, among those with non-perforated PSDs, terminals with multiple synaptic contacts had more mitochondria than those forming single synapses. PMID- 12536049 TI - Action potentials from human neuroblastoma cells in magnetic fields. AB - The patch-clamp method was used to measure transmembrane Na(+) and K(+) currents of the action potential in SH-SY5Y neuroblastoma cells exposed to static magnetic fields of 1, 5, and 75 G, 60 Hz fields of 1 and 5 G, and to combined static and low-frequency fields tuned for resonance of Na(+) and K(+). The maximum currents and their inactivation rates, and the activation rate of the Na(+) current were measured. Application of the magnetic fields did not result in detectable changes in any of the parameters of the action potential chosen for study. The occurrence of effects due to the fields could be excluded down to at least one part in 1000. The results suggest that magnetic fields of the type studied do not affect the cellular mechanisms responsible for generating the action potential. PMID- 12536050 TI - Inflammation induced increase of fluoride resistant acid phosphatase (FRAP) activity in the spinal dorsal horn in rats. AB - Fluoride-resistant acid phosphatase (FRAP) has been suggested as an enzymatic marker for nociceptive primary afferent terminals in the spinal dorsal horn, however there has not been demonstrated a direct functional relation between FRAP activity and an increased nociceptive transmission. For this purpose, we quantitated FRAP activity in the spinal dorsal horn of the rat in a heat-induced cutaneous inflammatory model. Male Sprague-Dawley rats anaesthetised with thiopental were separated in two groups where the left hindpaw was submerged during 60 s either in water at room temperature (control group) or in water at 60 degrees C (inflammation group) which induce in this group a progressive hindpaw inflammation. After 8 h, the lumbar enlargement of the spinal cord was extracted, cut in slices and 1 mm micropunch fragments were obtained from the right and left dorsal horn. The activity of FRAP was determined using the Gomori colorimetric method and corrected by the protein concentrations. FRAP activity in the left dorsal horn was statistically higher than right dorsal horn in the inflammation group (3.05+/-0.54 versus 1.91+/-0.23 u/g per l; P<0.05). Also, FRAP activity from the left dorsal horn of the control and inflammation groups show a significant increase in the last group (3.05+/-0.54 versus 2.17+/-0.23 u/g per l; P<0.05). This results demonstrate that FRAP is not only an enzymatic marker for neuronal and fibre integrity of nociceptive primary afferents but also it is associated to the nociceptive afferent activation. PMID- 12536051 TI - Epilepsy and pregnancy: Report of an Epilepsy Research Foundation Workshop. AB - Pregnancy in women with epilepsy (WWE) is known to be associated with a higher risk of congenital malformations than is associated with pregnancy in non epileptic women. Several factors have been identified to account for the increased risk, including the direct teratogenic effects of antiepileptic drug (AED) therapy, indirect effects of these drugs by interfering with folate metabolism, genetic abnormalities in drug or folate metabolism, and possibly an arrhythmogenic effect of maternal drug therapy on the embryonic heart, leading to ischaemia in developing tissues. A harmful effect of maternal seizures on the developing embryo has not been proven, although seizures and status epilepticus account for most of the excess maternal mortality in women with epilepsy. Abrupt withdrawal of drug therapy by the mother may be an important contributory factor. Less is known about the psychomotor development of children born to mothers with epilepsy because few studies have been designed to follow their progress throughout childhood. Retrospective studies suggest that impaired cognitive development may be associated with maternal drug therapy, particularly valproate. There is an urgent need to evaluate these risks and, with this in mind, several prospective registers have been set up to collect data from pregnancies in women with epilepsy. PMID- 12536052 TI - Safety and tolerance of rapidly infused Depacon. A randomized trial in subjects with epilepsy. AB - BACKGROUND: Valproate sodium injection (Depacon(R)) is an intravenous form of valproate for use in absence and complex partial seizures when circumstances preclude oral administration. Certain situations may warrant larger and more rapid infusions than permitted by the original labeling. This study evaluated the safety of more rapid infusions. METHODS: Subjects with epilepsy were randomized in a 2:1 ratio to receive up to 15 mg/kg of valproate sodium infused at 3.0 or 1.5 mg/kg/min. Up to four infusions were allowed within 24 h to achieve target plasma valproate concentrations of 50-100 mcg/ml. Primary safety endpoints were the changes in the 5-min and minimum post-first infusion blood pressures (BPs). RESULTS: One hundred twelve subjects were treated, (3.0 mg/kg/min group: n=72, 1.5 mg/kg/min group: n=40). No significant treatment differences were detected for changes in the primary BP endpoints. Two subjects in the 3.0 mg/kg/min group had potentially clinically significant low systolic BP values during the study. Similar proportions of subjects in the two groups reported adverse events during or within 6 h following the first infusion. CONCLUSIONS: Valproate sodium injection dosages up to 15 mg/kg and rates of 1.5 and 3.0 mg/kg/min were well tolerated in this population. PMID- 12536053 TI - alpha-[11C]-Methyl-L-tryptophan PET identifies the epileptogenic tuber and correlates with interictal spike frequency. AB - Epilepsy surgery has been successfully performed in patients with tuberous sclerosis complex (TSC) and seizures arising from a restricted epileptogenic area. The outcome of cortical excision depends on accurate pre-surgical identification of the epileptogenic tuber. [11C] alpha-methyl-L-tryptophan (alpha MTrp) was originally developed to measure serotonin synthesis in vivo with positron emission tomography (PET). However in pathologic conditions its uptake may also depend on the synthesis of quinolinic or kynurenic acid via the kynurenine pathway. Increased levels of serotonin and quinolinic acid have been observed in epileptogenic lesions, raising the possibility that alpha-MTrp PET may localize the epileptogenic area. The aim of this study was to correlate alpha MTrp PET uptake with the localization of the epileptogenic area and with interictal spike frequency in patients with TSC. alpha-MTrp uptake was measured in 8 patients (2 males, mean age 29.6+/-14.9 years, range 3-50 years) with intractable partial epilepsy due to TSC. All patients underwent scalp EEG monitoring during the PET scan. In four (50%), increased uptake of alpha-MTrp occurred in the epileptogenic area alone. Two (25%) patients showed multifocal abnormalities and the remaining two (25%) did not show focal changes. PET localization was mostly seen in patients with frequent interictal abnormalities on the EEG. Furthermore, there was a significant correlation between alpha-MTrp uptake and the frequency of interictal spikes (r=0.6; P<0.05). alpha-MTrp PET is a promising diagnostic tool in the localization of the epileptogenic area in patients with TSC. PMID- 12536054 TI - Differential effects of dihydropyridine calcium channel blockers in kainic acid induced experimental seizures in rats. AB - The anticonvulsant effects of the dihydropyridine calcium channel blockers nifedipine, nicardipine and nimodipine were studied on experimental seizures induced by intra-hippocampal injection of kainic acid (KA) in chloralhydrate anesthetized Wistar rats. The rats were anesthetized and placed in a stereotaxic apparatus. After midline incision four screw electrodes were placed over the left and right frontal and parietal cortex and KA was injected into left dorsal hippocampus via 5-microliter Hamilton microsyringe. The changes in electroencephalograph (EEG) activity and EEG power spectra were recorded in basal conditions and 5, 10, 15, 30 and 60 min following KA injection. KA-induced excitatory changes in the surface EEG activity were associated with the marked increase in EEG power spectra in the frequency range from 14.5-22 Hz. Pretreatment with nifedipine, nicardipine and nimodipine revealed that they exerted certain differences in their anticonvulsant properties. Nimodipine significantly delayed the onset of seizures and prevented the KA-induced changes in EEG and in EEG power spectra in all recorded channels and in a dose dependent manner. Nifedipine exerted significant anticonvulsant effect only in channel four, while nicardipine was ineffective. Our results suggest that anticonvulsant action of some dihydropyridine calcium channel blockers, especially nimodipine may be in part independent of its antagonism on L-type voltage-gated calcium (Ca(2+)) channels. PMID- 12536055 TI - Anxiety as a differential factor in epileptic versus psychogenic pseudoepileptic seizures. AB - Anxiety may be a hypothetical factor responsible for psychogenic pseudoepileptic seizures. The purpose of this study was to analyse anxiety indexes manifested in the Minnesota Multiphasic Personality Inventory. Patients were divided into three groups on the basis of a neurological examination and long-term video-monitoring. Group One (N=70; 58 F, 12 M) had only psychogenic pseudoepileptic seizures. Group Two (N=40; 31 F, 9 M) had both epileptic seizures and psychogenic pseudoepileptic seizures. Group Three (N=42; 30 F, 12 M) had only epileptic seizures and served as the control group. Patients with psychogenic pseudoepileptic seizures, alone or in combination with epileptic seizures scored significantly higher than the epileptic group on the following anxiety measures: Anxiety Sign (Gough) - AxS - (P<0.001), Expressive-repressive Index (Sandford, Webster and Freedman) - ERI (P<0.001), and the Neurotic score (Ruesch and Bowman) - NS (P<0.001). Difference between all three groups were found for the Triad Elevation Index (Lovell)-TI (P<0.001) and the Frustration Tolerance Index (Beall and Panton) - FT (P<0.005). These findings suggest that the existence of psychogenic pseudoepileptic seizures or the predisposition to such seizures is reflected in the anxiety dimensions of the personality profile. Psychological evaluation of anxiety may help us to gain a better understanding of, and discrimination between, patients with psychogenic pseudoepileptic seizures, mixed seizures and epileptic seizures. PMID- 12536056 TI - Prevalence of adult epilepsy in Estonia. AB - Epidemiological data about epilepsy from central and eastern Europe is scarce and results are controversial. The aim of this study was to estimate the main prevalence-related characteristics of active epilepsy in an adult population in Estonia. Review of all databases and lists related to epilepsy in Tartu supplemented by re-examination of patients to identify all persons with active epilepsy aged >/=20 years on January 1, 1997. Special attention was paid to the extensive adoption of definitions and criteria proposed by ILAE guidelines for epidemiologic studies. Both, crude and age-adjusted (to the 1970 US population) prevalence rates were 5.3 per 1000. The age-specific rates were constant in age groups 30-69 years and declined in the oldest age groups. Of the seizure types, partial seizures had highest rates, over half were secondarily generalized seizures. The largest syndromic categories were localization-related symptomatic and cryptogenic epilepsies. Risk factors for epilepsy were identified in 39.6% cases. Some 22% of all subjects did not take antiepileptic medication. Prevalence of active epilepsy and other prevalence-related characteristics in the adult population of Tartu was comparable to those reported from the developed countries. The predominance of localization-related syndromes and partial seizures is due to the age distribution of the study. PMID- 12536057 TI - Unaltered control of extracellular GABA-concentration through GAT-1 in the hippocampus of rats after pilocarpine-induced status epilepticus. AB - The uptake of the inhibitory transmitter GABA (gamma-aminobutyric acid) limits the efficacy of synaptic and tonic inhibition in brain tissue. It has been reported that GABA-uptake is down-regulated in temporal lobe epilepsy. This down regulation may increase the inhibitory action of GABA but may also limit the anticonvulsant activity of GABA-uptake blockers. We have directly compared the function of GABA-uptake in hippocampal slices from normal and chronically epileptic rats. We raised the global extracellular concentration of GABA by bath application of the agonist in the absence and presence of the GABA-uptake blocker tiagabine. GABA-induced currents were measured in dentate granule cells and CA1 pyramidal neurons in hippocampal slices. The potentiation of currents by tiagabine was taken as a measure for the efficacy of GABA-uptake in the hippocampal tissue. There was no difference between cells from control- or pilocarpine-treated animals in the response to GABA or in the conductance increase following application of tiagabine. Our data show that in the chronic phase of the pilocarpine-model GABA-uptake maintains its ability to control the extracellular background concentration of GABA. PMID- 12536058 TI - Effects of remacemide in two models of genetically determined generalized epilepsy, the GAERS and the audiogenic Wistar AS. AB - The antiepileptic effects of remacemide were assessed in two models of genetically determined generalized epilepsy. The model of non-convulsive epilepsy used was a model of absence seizures, the GAERS (genetic absence epilepsy rats from Strasbourg), and the model of convulsive seizures was an audiogenic rat model, the Wistar AS. In the eight GAERS studied, the three doses of remacemide (20, 40, and 80 mg/kg) dose-dependently reduced the expression of spike-and-wave discharges (SWDs) that had almost totally disappeared at the highest dose used, 80 mg/kg. However, at the latter dose, the effect of remacemide may be partly due to a change in the vigilance level of the animals. In the Wistar AS, the dose of 20 mg/kg prolonged by twofold the latencies to wild running and tonic seizures, and prevented their expression in one rat out of the eight studied. At 40 mg/kg, the expression of wild running and tonic seizures was inhibited in seven and maintained in one of the eight rats studied. The present results support the effects of remacemide in tonic/clonic seizure, which was the first target of the drug, and confirm the effect of the anticonvulsant on absence seizures. PMID- 12536059 TI - Suppression of epileptiform activity by GABA(B) receptors in wild type and weaver hippocampus 'in vitro'. AB - Inhibition by GABA(B) receptors comprises activation of K(+) conductance and inhibition of Ca(2+) conductance, thereby reducing action potential dependent transmitter release and silencing neuronal activity. We compared epileptiform activity and its inhibition by the activation of GABA(B) receptors in homozygous weaver (wv/wv) and wild type (+/+) CA3 neurons disinhibited by GABA(A) receptor blockade. In wv/wv mice GABA(B) receptors have lost their ability to activate K(+) conductance (J. Neurosci. 18 (1998) 4001). Spontaneous synchronous burst discharges in elevated [K(+)](o) displayed only subtle differences in +/+ and wv/wv slices, except that the GABA(B) receptor agonist R-baclofen in low concentration (0.1 microM) strongly reduced the frequency of synchronous bursts in +/+ CA3 neurons, but not in wv/wv CA3 neurons. A high affinity GABA(B) antagonist, CGP55845A (0.5 microM) promoted the incidence of bursts in low [K(+)](o). Concentration dependence of the reduction of evoked EPSCs was identical in wv/wv and +/+ neurons (IC(50)=0.3 microM). Amplitudes of evoked IPSCs were reduced by 0.01 microM R-baclofen in +/+, but not in wv/wv CA3 neurons. The effect of the low concentration was abolished by Ba(2+), which is known to block Kir conductance. The data suggest that activation of Kir conductance is important for the control of GABA release by GABA(B) autoreceptors in the CA3 network. We conclude that the loss of a contribution of Kir conductance to GABA(B) receptor-mediated autoinhibition reduces the inclination towards spontaneous bursts of wv/wv CA3 pyramidal neurons. PMID- 12536060 TI - Concurrent treatment for alcohol and tobacco dependence: are patients ready to quit both? AB - The prevalence of smoking among alcohol abusers is high, yet little is known about this dual-dependency. This study examines mechanisms involved in changing both alcohol and tobacco use concurrently using the transtheoretical model (TTM) measures of change. Alcohol and tobacco dependent outpatients (N=115) entering a dual-substance dependence program were compared on baseline measures of motivation, self-initiated change activities, and self-efficacy associated with each substance use behavior. Differences on these measures were expected for drinking versus smoking. Motivation to change each behavior was also examined as a potential predictor of retention in treatment. Results indicated that patients reported higher self-efficacy to abstain and lower temptation to use alcohol relative to cigarettes. Change activities were also initiated at higher levels for drinking compared with smoking. An interaction between drinking and smoking motivation for change was found in the prediction of treatment retention; those with higher motivation for changing their alcohol use and lower motivation to quit smoking remained longer in treatment, while those who were higher in motivation for changing both behaviors dropped out the earliest. Overall, participants in this dual-dependence program were more confident and active in changing their alcohol use. Initiating cessation of both behaviors equally and simultaneously may prove difficult for this population. This study initiates an understanding of the mechanisms involved in changing alcohol-tobacco dependence and may provide guidance for developing dual cessation interventions. PMID- 12536061 TI - Primary health care professionals' activity in intervening in patients' alcohol drinking during a 3-year brief intervention implementation project. AB - AIM: The study examined the change of primary health care professionals' activity in asking patients about alcohol and giving advice to heavy drinkers during a 3 year brief intervention implementation project. METHOD: A patient questionnaire survey after consultation blind to the primary health care professionals before and after the implementation project. Brief intervention implementation was based on action research. Subjects were 1,000 consecutive 16-65-year-old patients consulting general practitioners at two primary health care centres at baseline and follow-up, of whom 655 and 768, respectively, participated. RESULTS: No statistically significant differences were found regarding asking or advising about alcohol between baseline and 3-year follow-up. Of all patients, 19.1% (125/655) at baseline versus 19.7% (151/768) at follow-up were last asked about alcohol during the consultation in question or during past year (P=0.784). Likewise, of heavy drinkers, 30.9% (30/97) versus 33.9 (38/112) were asked (P=0.644). Of heavy drinkers, 13.4 (13/97) versus 14.9% (17/114) were advised about alcohol during the consultation in question (P=0.754). CONCLUSION: The activity to do brief intervention remained stable. This may be due to the short follow-up time, the way brief intervention was implemented in the present study or the saturation in the activity reached already before the present study. PMID- 12536062 TI - Socio-economic status and binge drinking in Israel. AB - Modern Israeli society is comprised primarily of two nationality groups-Jews and Arabs, with disparate religious and cultural attitudes toward alcohol drinking. We recently described higher rates of past-month drinking among Jewish adults, although Arabs who drink were more likely to report binge drinking. The goal of the present study is to examine the relationship between binge drinking and socio economic status (SES) among Arab and Jewish adults in Israel. Data from a 1995 nationally representative household survey on drug and alcohol use were analyzed. Participants included male and female Arabs (n=982) and Jews (n=4,972) aged 18-40 living in Israel. SES was assessed using education, household income, and occupation. The prevalence of binge drinking was highest among Arab men (21.4%) followed by Jewish men (15.2%), Arab women (7.3%), and Jewish women (4.0%). Prevalence rates and odds ratios (ORs) from logistic models controlling for age, gender, marital status and religiosity show that increased household income and occupation are associated with increased binge drinking among Arabs (OR>2.0) and decreased binge drinking among Jews (OR congruent with 0.6). Higher educational achievement was protective against binge drinking in both nationality groups. Varied results for income and occupation, and education indicate the need to examine the association between each SES indicator and alcohol consumption independently, especially in culturally diverse populations. PMID- 12536063 TI - Patterns of use and harms associated with non-medical ketamine use. AB - AIM: To (1) identify current patterns of non-medical ketamine use; and (2) identify potential harms associated with non-medical ketamine use. DESIGN: Cross sectional survey of lifetime ketamine users. SETTING: Semi-structured interviews took place in public and private settings in Sydney Australia. PARTICIPANTS: One Hundred ketamine users. MEASUREMENTS: Self-reported experiences with and attitudes towards ketamine use. FINDINGS: Ketamine appeared to be added to an already extensive drug use repertoire of a well-educated and informed sample. Many users reported regularly experiencing effects such as an inability to speak, blurred vision, lack of co-ordination and increased body temperature, which resulted in some either reducing their dose or stopping use. CONCLUSIONS: Many users had experienced significant negative effects, such that some had either reduced their dose or stopped use altogether and expressed concerns over some others. This study reinforces the need to develop harm minimisation campaigns that match the experiences and attitudes of their target group through careful needs assessment and appropriate evaluation. PMID- 12536064 TI - Drug use among two American Indian populations: prevalence of lifetime use and DSM-IV substance use disorders. AB - American Indians (AIs) have often reported higher rates of drug use than have other racial/ethnic groups. However, the majority of these studies have focused on drug use among high school adolescents, with little attention to pathological use such as drug abuse or dependence. This study is among the first to report lifetime drug use and disorder (abuse/dependence) information from community samples of two culture groups of AI people-one in the Southwest (SW), one in the Northern Plains (NP)-ranging in age from 15 to 57 years old. Analyses were conducted within four groups: SW men, SW women, NP men, and NP women. Across the four groups, lifetime use rates for marijuana (36.9-57.5%), cocaine (4.3-21.5%), and inhalants (3.6-17.0%) were the highest drug use rates; heroin (0.5-2.1%), the lowest. Lifetime drug disorder rates were highest for marijuana (4.5-14.1%), cocaine (1.1-2.3%), and stimulants (0.7-1.7%). Lifetime polydrug use disorder rates from 1.2 to 4.5%. Women generally had lower prevalence rates than did men in their culture group. The SW women generally had the lowest rates of use and disorder. Lifetime use and disorder rates among the youngest group were often not different from rates of the older groups. Overall, 40-60% had never used any drugs; 85-95% had not developed any drug disorder. Despite widespread concern and rhetoric about drug problems among AIs, many who had used various drugs either were using them without serious consequences or had quit use altogether. PMID- 12536065 TI - A comparison of African American and non-Hispanic Caucasian cocaine-abusing outpatients. AB - An understanding of how different racial and ethnic groups vary with respect to demographic characteristics and severity of psychosocial problems is necessary to better address the treatment needs of these subgroups. This study evaluated similarities and differences between African American and non-Hispanic Caucasian cocaine users initiating substance abuse treatment at community-based programs. At intake to treatment, patients completed the addiction severity index (ASI). African Americans (n=161) and Caucasians (n=86) differed with respect to gender, income and age. After controlling these demographic variables and treatment site, race-related differences were noted in many of the ASI composite scores. While African Americans had more severe employment problems, they had less severe alcohol, legal, family/social and psychiatric difficulties than Caucasians. These differences suggest that African Americans and Caucasians may benefit from treatment emphasizing different aspects of their psychosocial problems. Specifically, for many African American patients, employment-training programs may be particularly useful. Caucasians may be in greater need of treatment that focuses on concurrent cocaine and alcohol problems, as well as family and psychiatric difficulties. PMID- 12536066 TI - Physical and sexual abuse, depression and alcohol use disorders in adolescents: onsets and outcomes. AB - Adolescents with alcohol use disorders (AUDs) often have major depressive disorder (MDD). While physical abuse and sexual abuse (PS Abuse) have been observed to be common in adolescents with AUDs, the influence of PS Abuse on comorbid MDD and AUD has not been determined. The effect of pre-existing PS Abuse on the young adulthood outcomes of adolescents with AUDs has also not been adequately explored. This study examined the relationships among PS Abuse, MDD, and AUD in adolescence, as well as related young adult outcomes. Adolescents (mean age: 16.4 years; range: 14-18 years) were recruited from clinical and community sources and classified into four groups: (1) AUD+PS Abuse (n=154), (2) AUD only (n=255), (3) PS Abuse only (n=74), and (4) Controls (n=268). Subjects were longitudinally assessed through young adulthood (age 19 years or older). Measures included interview assessments of DSM-IV AUD and MDD, classified as "primary" or "secondary", and questionnaire measures of alcohol consumption and depression. Primary MDD preceded AUD whereas secondary MDD had a later onset than AUD. PS Abuse accelerated the onsets of primary MDD, secondary MDD and AUD. While affected adolescents had typically improved in both alcohol consumption and depression at the young adult assessment, the majority of those with adolescent AUD had AUDs in young adulthood, and MDD remained common in those with a history of PS Abuse. These results indicate that MDD among adolescents with AUD may be partly attributable to PS Abuse. PMID- 12536067 TI - Changes in smoking status among substance abusers: baseline characteristics and abstinence from alcohol and drugs at 12-month follow-up. AB - The impact of change in smoking status on 12-month substance abuse (SA) treatment outcomes was examined among an HMO population seeking SA treatment. Of the 749 participants who entered the study at baseline, 649 (86.9%) were retained at the 12-month follow-up. At treatment entry, 395 participants were smokers and 254 were nonsmokers. At 12-month follow-up, 13% of the 395 baseline smokers reported quitting smoking and 12% of the 254 baseline nonsmokers reported starting/relapsing to smoking. Those who quit smoking were less likely to be diagnosed as alcohol dependent compared to those that remained smokers. Those who started/resumed smoking were more likely to be diagnosed as both alcohol and drug dependent at treatment entry compared to all other groups. Total days abstinent from alcohol and illicit drugs was greatest for individuals who quit smoking (adjusted M=310.6) or who were nonsmokers (adjusted M=294.7) and lowest for those who started/resumed smoking (adjusted M=246.6) or remained smokers (adjusted M=258.2), even after controlling for demographic (i.e. age, income), psychosocial (ASI psychiatric severity), and other treatment characteristics (length of treatment stay, prescribed bupropion) that were associated with days abstinent at 12 months. Self-initiated smoking cessation does not appear to be detrimental to SA treatment outcomes, and may be beneficial. Starting/resuming smoking after entering SA treatment may be a clinical marker for individuals at greater risk of relapse. Future studies may want to measure the smoking status of all participants at all time points in order to include this higher-risk group of substance using smokers. PMID- 12536068 TI - What does it take to retain substance-abusing adolescents in research protocols? Delineation of effort required, strategies undertaken, costs incurred, and 6 month post-treatment differences by retention difficulty. AB - Research retention rates vary widely due to practical difficulties that can be exacerbated when participants are minors. This article describes: (1) the range of effort required and type of follow-up strategies used to complete face-to-face follow-up interviews with substance-abusing adolescent research participants; (2) common locations of follow-up interviews; and (3) characteristics of difficult- versus easy-to-retain adolescent participants. Diverse contact strategies and numerous contact attempts were needed to obtain a 94% 1-month and 92% 6-month retention rate among substance-abusing adolescent research participants. About half of the youth did not respond to basic telephone tracking and required enhanced tracking efforts. Approximately 40% of the youth required 6 or more contacts prior to interview completion. The majority of follow-up interviews (60%) were conducted in community settings such as fast food restaurants, constituting the adolescent's preferred interview location. Telephone interviews were infrequent since adolescents wanted privacy and were concerned that a household member would listen to their answers. Those youth proving difficult-to retain were significantly more likely to report serious problem behavior and poorer outcomes 6-months post-treatment within the alcohol/drug, juvenile justice, family, and educational domains. It was estimated that an additional $85 per participant per follow-up wave (over and above project budgets) was needed to adequately track, locate and interview an adolescent research participant. This expenditure appears reasonable to ensure a reliable/valid data set. Assessing the cost/benefit of different methods used in preventing attrition, identifying the minimum standards that avoid response bias and examining the impact of interviewer/participant alliances on data reliability/validity is discussed. PMID- 12536069 TI - Agreement between head of household informant and self-report in a community survey of substance use in India. AB - This survey of 500 households in a New Delhi urban slum compared reports of substance use by the head of the household informant with individual self-report. Information from the two sources was compared for 1,132 people above the age of 15 years. The paired agreement regarding the use of substances was high (kappa=0.92; S.E.=0.01, z=92.0). The agreement regarding the presence of symptoms and classification of dependence for the use of alcohol, tobacco and opiates ranged from good to excellent and head of household reports had a high positive predictive value for the use of these substances. This method provides useful estimates of drug use and dependence for substances associated with observable physiologic withdrawal syndromes, and is less costly and quicker to perform than traditional self-report methodologies. PMID- 12536070 TI - Comparing homeless and domiciled pregnant substance dependent women on psychosocial characteristics and treatment outcomes. AB - The present study compared pregnant drug-dependent women reporting homelessness (n=117) or being domiciled (n=118) at treatment enrollment on initial psychosocial functioning and subsequent drug treatment outcome. Homeless pregnant women presented with greater drug use and medical problems, less social service income, and more family/social difficulties than domiciled women. Homeless women had greater rates of psychiatric problems including suicidality and higher rates of physical, emotional and sexual abuse than their domiciled counterparts. Treatment retention was poorer for homeless than domiciled pregnant women. Results suggest that treatment should address the housing needs of pregnant drug abusers as well as their related social and psychiatric problems. PMID- 12536072 TI - Effect of separate conjugated linoleic acid isomers on murine mammary tumorigenesis. AB - Recent studies have linked conjugated linoleic acid (CLA) to altered tumorigenesis of several sites. We showed recently that a mixture of CLA isomers was able to significantly decrease mammary tumor metastasis in mice. That effect was seen with as little as 0.1% CLA in the diet. Other studies with dietary CLA have shown that various isomers may have differential effects. The purpose of this work was to assess which individual CLA isomers had similar effects in alteration of mouse mammary tumor metastasis. For that, we fed six 20% (w/w) total fat diets which contained either no CLA, low (0.1%, w/w) or high (0.25%, w/w) levels of cis9,trans11-CLA (c9,t11), trans10,cis12-CLA (t10,c12) or a mixture of the 2 isomers (0.125% of each, w/w) as free fatty acids. Neither the separate isomers nor the mixture had an effect on the latency or growth of primary line 4526 tumors when compared to the group without CLA. However, all diets containing CLA significantly decreased the total tumor burden (volume of tumor, mm(3)) in lungs of mice from both spontaneous metastasis (reduced by 42 73%) as well as implantation and survival of the metastatic cell (reduced by 46 61%) when compared with diets containing no CLA. Diets containing a greater concentration of either c9,t11 or t10,c12 had a significantly greater effect compared to the lower concentrations of the respective isomers when metastatic nodule size and total tumor burden were assessed. The diet containing both isomers decreased total tumor burden similarly to the diets containing the lower concentration of each of the isomers. Thus, the effects of c9,t11 and t10,c12 may not be additive and possibly share similar mechanisms for decreasing metastatic tumor burden in mice transplanted with mammary tumor cells. PMID- 12536071 TI - Histidine kinases and histidine phosphorylated proteins in mammalian cell biology, signal transduction and cancer. AB - Intensive investigation of protein tyrosine, serine and threonine phosphorylation has lead to advances in signal transduction research and cancer treatment. This feature summarizes research on mammalian proteins exhibiting histidine phosphorylation. Histidine kinases are well known in prokaryotic and lower eukaryotic systems where they form the 'two-component' signal transduction system. The relative invisibility of histidine phosphorylation in mammalian cells may result from technical obstacles such as its acid lability, which precludes detection in electrophoretic systems, amino acid sequencing, etc. Emerging data have identified mammalian histidine kinases for the kinase suppressor of ras, a scaffold molecule for the Map kinase pathway, as well as histone H4, aldolase C and the beta-subunit of heterotrimeric G proteins. Additional mammalian proteins of interest to signal transduction and cancer research exhibit histidine phosphorylation, including P-selectin, annexin I and the 20S proteasome. Other candidate histidine phosphorylated proteins are identified. These data suggest the existence of another series of phosphorylation patterns in signal transduction. PMID- 12536073 TI - Prolonged effects of beta-estradiol 3-benzoate on thyroid tumorigenesis in gonadectomized rats pretreated with N-bis(2-hydroxypropyl)nitrosamine. AB - The prolonged modulatory effects of beta-estradiol 3-benzoate (EB), a synthetic estrogenic compound, were investigated in a rat two-stage thyroid tumorigenesis model. One week after a single subcutaneous (s.c.) injection of N-bis(2 hydroxypropyl)nitrosamine, gonadectomized F344 rats of both sexes were s.c. implanted with fused pellets containing EB for 32 weeks. Doses of EB at 0, 0.004, 0.02 and 0.1mg were achieved by varying the ratio of EB to cholesterol in the pellet. Major organs including the thyroid, pituitary, liver, kidneys, uterus and brain were weighed and histopathological observation was performed. Serum was assayed for triiodothyronine (T(3)), thyroxine (T(4)) and thyroid-stimulating hormone (TSH). Thyroid weights were increased by the EB pellet implantation in a dose-dependent manner and significantly (P<0.05) elevated in the 0.1mg EB male group and in the 0.02 and 0.1mg EB female groups. The EB treatments dose dependently suppressed serum T(4) levels and inversely elevated serum TSH levels in both sexes but without statistical significance in females. Histopathologically, EB increased the occurrence of thyroid proliferative lesions in males and showed a tendency for increase in females. Interestingly, the effect of EB was more intensive in males than in females, even the lowest dose inducing a follicular carcinoma in a male. These results, thus indicate the possible contribution of prolonged EB stimulation at lower doses to thyroid tumorigenesis without additional promotive condition. PMID- 12536074 TI - LY294002, an inhibitor of phosphatidylinositol-3-kinase, causes preferential induction of apoptosis in human multidrug resistant cells. AB - Protein kinase B (PKB), a kinase downstream of phosphatidylinositol 3-kinase (PI3 kinase) provides anti-apoptotic and survival signals via phosphorylation of various targets. Inhibiting PI3-kinase with a 12 h exposure to 10 microM LY294002 induces levels of apoptosis of 30.39+/-1.53% in the KB-V1 multidrug resistant (MDR) cell line compared to 4.54+/-1.00% in drug sensitive KB-3-1 cells (P<0.001). This occurred in conjunction with a preferential reduction in activated PKB in MDR cells. These results suggest the PI3-kinase/PKB signalling pathway is important for the survival of MDR cells and inhibition of this pathway results in the selective induction of apoptosis in MDR cells. PMID- 12536075 TI - Glutathione S-transferase polymorphisms in thyroid cancer patients. AB - Glutathione S-transferases (GST) are enzymes involved in the metabolism of many carcinogens and mutagens, also acting as important free-radical scavengers. The existence of different genetic polymorphisms in human populations has proven to be a susceptibility factor for different tumours. Nevertheless, as far as we know, for thyroid cancer no study has been conducted until now linking its incidence to genetic susceptibility biomarkers. The present investigation has been conducted to detect the possible association between polymorphism at the GSTM1, GSTT1 and GSTP1 genes and thyroid cancer incidence. Thus, 134 thyroid cancer patients and 116 controls, all from the urban district of Barcelona (Spain), have been included in this study. The results indicate that, according to the calculated odds ratio, the frequencies of the different genotypes found in the group of cancer patients do not significantly differ from those values obtained in the controls. This is true for the overall data as well as for the tumour characterization as follicular and papillar types. In addition, none of the possible combinations of mutant genotypes were shown to be risk factors. Finally, when the sex of the patients, the age of tumour onset, and life-style habits were also taken into account, no influence was observed related to the different genotypes. In conclusion, the results obtained in this study clearly suggest that those susceptibility factors related to the different GST polymorphic enzymes are not a predisposing factor in thyroid cancer disease. PMID- 12536076 TI - Myenteric denervation reduces the incidence of gastric tumors in rats. AB - The influence of myenteric denervation on the development of gastric tumors induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) was studied after chemical denervation of the rat stomach with benzalkonium chloride (BAC). Three groups were evaluated: control, denervated and denervated with pyloroplasty. Random bred male Wistar rats were given MNNG in drinking water (100 mg/l) for 28 weeks. After the sacrifice of animals, the stomachs were removed for morphological study. BAC reduced myenteric neurons number, increased the gastric mucosa area and decreased the adenocarcinomas number and size. This decrease was more evident when denervation was associated with pyloroplasty. These results indicate that myenteric denervation reduces the incidence of experimentally induced gastric tumors. PMID- 12536077 TI - Anti-proliferative effects of 20-epi-vitamin-D3 analogue, KH1060 in human neuroblastoma: induction of RAR-beta and p21(Cip1). AB - We determined the in vitro biological activities of 1 alpha, 25-dihdroxyvitamin D(3) (1,25-D(3)) and its analogue, 20-epi-22-oxa-24a, 26a, 27a-trihomo-1 alpha, 25 (OH)(2) vitamin D(3) (KH1060) in six human neuroblastoma (NB) cell lines (SH SY5Y, NB69, SK-N-AS, IMR5, CHP-134, NGP). The ability of these compounds to inhibit cell growth and DNA synthesis was evaluated by 3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyl tetrazolium bromide assay and BrdU incorporation, respectively. The induction of cell death was monitored by caspase-3 activity. Their antineoplastic effect was assessed by clonal proliferation in soft agar. KH1060 was more effective than 1,25 D(3) in inhibiting cell growth and DNA synthesis. The IC-(50) (inhibition of 50% cell viability) indicated that KH1060 was about 10 20-fold more potent than 1,25 D(3). This growth inhibition was also accompanied by induction of caspase-3 activity, indicating that these compounds induce cell death in a caspase-dependent fashion. Moreover, KH1060 exerted potent antineoplastic activity by suppressing the clonal proliferation of the six NB cells. For the first time we demonstrate that KH1060 induces the expression of retinoic acid receptor-beta and p21(Cip1) suggesting that these proteins in part mediate the growth inhibitory effects. Taken together, all the six NB cells were more susceptible to growth inhibition by KH1060 than 1,25-D(3), suggesting its possible use in NB to potentiate the action of retinoids, which are in clinical use for this disease. PMID- 12536078 TI - Physiological and molecular effects of Apo2L/TRAIL and cisplatin in ovarian carcinoma cell lines. AB - Combining of tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL) with a chemotherapeutic drug, cisplatin, in ovarian carcinoma cell lines exerted potent anti-tumor effects that exceeded the effects of each drug alone. In order to investigate mechanisms of anti-tumor activity of cisplatin/Apo2L/TRAIL combination, we assessed in detail the molecular effects of cisplatin and Apo2L/TRAIL-activated cell death in two ovarian carcinoma cell lines, OVCAR3 and SKOV3, using cDNA array hybridization, Western blot and flow cytometry. We observed differential induction of apoptosis-related molecules by cisplatin and Apo2L/TRAIL. Cisplatin upregulated the expression of both death and decoy TRAIL receptors, as well as of TRAF5 and -6, downregulated the anti apoptotic proteins, Bcl-2, and induced activation of caspases-3, -8 and -9. Apo2L/TRAIL induced the expression of pro-apoptotic proteins, Bad and Bax; downregulated the anti-apoptotic proteins, Bcl-2 and Bcl-xL; and activated caspases-3, -7, -8, -9 and -10. Cisplatin/Apo2L/TRAIL combination resulted in further downregulation of expression of anti-apoptotic proteins, Bcl-2 and Bcl xL, as well as an increase in mitochondrial permeability transition and activation of caspases-3, -8, and -10. These data demonstrate positive cooperation of cisplatin and Apo2L/TRAIL and emphasize the potential clinical usefulness of cisplatin/Apo2L/TRAIL combination therapy. PMID- 12536079 TI - Mutations of the D310 mitochondrial mononucleotide repeat in primary tumors and cytological specimens. AB - A mononucleotide repeat (D310) in mitochondrial DNA has been recently identified as a mutational hot spot in primary tumors. We analyzed 56 tumors for insertion/deletion mutations in the D310 repeat. A total of 13 mutations were detected. The highest frequency of mutations was found for cervical cancer, followed by bladder tumors, breast cancer and endometrial neoplasia. No alterations were observed in four patients suspected of malignancy but without evidence of malignant tumor. We detected identical changes in four of four urine sediments from patients with bladder cancer and in three of three fine needle aspirates of patients with breast cancer. Our results indicate that D310 abnormalities are detectable in cytology specimens from patients with cancer and support the notion that D310 analysis may represent a new molecular tool for cancer detection. PMID- 12536080 TI - Characterization of the gene expression profile of neuroblastoma cell line IMR-5 using serial analysis of gene expression. AB - The serial analysis of gene expression (SAGE) technique was used to generate a database of the most abundant transcripts of the MYCN-amplified neuroblastoma cell line IMR-5. A total of 8568 tags were sequenced and shown to represent 4034 unique tags, each of which corresponds to an individual transcript. Expression levels of genes are reflected by the frequency of occurrence of the respective tags. To validate fidelity of SAGE data, relative abundances of seven transcripts were evaluated by semiquantitative reverse transcriptase-polymerase chain reaction. Transcripts that were detected nine times or more (>0.1% of the total tag population) accounted for 36% of the total messenger RNA mass but only 3% of the total number of individual transcripts. A strong preponderance of genes involved in protein synthesis, in particular those encoding for ribosomal proteins, were observed among these high-abundance transcripts. Tags corresponding to the amplified gene DDX1 were conspicuously overrepresented in comparison to the other amplified genes MYCN, neuroblastoma amplified gene and MEIS1, which suggests an additional mechanism apart from genomic amplification contributing to the strong upregulation of this gene. This study provides a comprehensive gene expression profile of neuroblastoma cell line IMR-5 and may be used as a reference database for identification of candidate genes that are involved in etiology and pathogenesis of neuroblastoma. PMID- 12536081 TI - Cyclin A1 is highly expressed in aggressive testicular germ cell tumors. AB - Cyclin A1 is a tissue-specific A-type cyclin that is essential for spermatogenesis. Overexpression of cyclin A1 was found in acute myeloid leukemia and cyclin A1 induced leukemia in a transgenic mouse model. We used quantitative real-time reverse transcription-polymerase chain reaction to analyze cyclin A1 expression in solid tumors. Cyclin A1 expression was very low in breast cancer, non-small cell lung cancer and in cervical carcinoma. However, substantial expression of cyclin A1 was found in testicular and ovarian cancer and in endometrial cancer. In testis specimens, cyclin A1 expression was much higher in testicular tumors compared to Sertoli cell only syndrome that lacks spermatogenesis. Compared to normal spermatogenesis, testicular cancers expressed on average lower levels of cyclin A1. Among the different histological subtypes of testicular tumors, embryonal cell carcinomas and immature teratomas expressed the highest levels of cyclin A1. The cyclin A1 levels in these tumors were similar to those seen in normal testis. Seminomas and yolk sac tumors expressed intermediate levels, whereas cyclin A1 expression was very low in mature teratomas. These findings indicate that cyclin A1 is expressed in selected solid tumors. Its known oncogenic function and the high expression levels in aggressive testicular tumors suggest a role for cyclin A1 in germ cell tumorigenesis. PMID- 12536082 TI - Expression of thymidylate synthase and thymidine phosphorylase in human breast carcinoma: implication for method to detect expression of these molecules in clinic. AB - Thymidine phosphorylase (dThdPase) is identical to platelet-derived endothelial cell growth factor, and has an angiogenic activity in experimental models and human solid tumors. A critical step in the de novo pathway of DNA synthesis is the production of the pyrimidine nucleotide dTMP from dump and this reaction is catalyzed by thymidylate synthase (TS). Both dThdPase and TS levels seemed to be related to response to 5-fluorouracil (5-FU) therapy in different types of human solid tumors. The present study evaluated dThdPase and TS expression levels by immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT PCR), using a same set of 39 breast carcinoma tissues for both methods. An inverted-relationship in the expression levels of TS and dThdPase was observed. Further, immunohistochemical analysis may be a better tool than analysis by RT PCR in detection of dThdPase and TS, because of both dThdPase and TS expression in cells besides carcinoma cells. These imply that immunohistochemical analysis of dThdPase and TS is available for selection of patients who will be received 5 FU based chemotherapy. PMID- 12536083 TI - Co-cultivation of pancreatic cancer cells with orthotopic tumor-derived fibroblasts: fibroblasts stimulate tumor cell invasion via HGF secretion whereas cancer cells exert a minor regulative effect on fibroblasts HGF production. AB - The intensive stromal reaction is one of characteristics of pancreatic exocrine carcinoma. The mutual interaction between pancreatic cancer cells and orthotopic tumor-derived fibroblasts have not been clarified yet. In this study, we sought to elucidate the mechanism underlying the tumor-stromal interaction with an in vitro coculture experimental system. Considerable strong c-Met expression was detected in seven out ten lines of human pancreatic carcinoma cells, as determined by Western blotting. For hepatocyte growth factor (HGF)-production, however, none or only trace amounts of HGF could be detected in those ten cell lines. Of the two lots of tumor-derived fibroblasts obtained from two pancreatic cancer patients, the fibroblasts capable to produce HGF could initiate an apparent invasion-stimulating response in strong c-Met-expressed Suit-2 and Panc 1 cells but not in faint expressed Mia PaCa-2 and BxPC-3 cells. A specialized HGF antagonist, NK4 would effectively inhibit the fibroblast-mediated invasive growth, thus proving the key role of the paracrine-fashioned HGF/c-Met pathway in the tumor-stromal interaction. On the other hand, the regulative action of cancer cells on HGF expression of fibroblasts was also investigated using direct or indirect coculture systems. For the fibroblasts that originally did not produce HGF, cancer cells failed to show any HGF-inductive effect. For the HGF-producing fibroblasts, despite of somewhat upregulation or downregulation in fibroblast HGF expression, the feedback regulation by studied pancreatic cancer cells in both coculture modes were relatively limited. This in vitro study sketched out the interaction between cancerous and stromal compartments with an emphasis on HGF/c Met signal pathway, thus possibly helping to unveil the more complicated mutual modulation in vivo between pancreatic cancer and host mesenchymal tissues. PMID- 12536084 TI - In vitro effects of estradiol, dydrogesterone, tamoxifen and cyclophosphamide on proliferation vs. death in human breast cancer cells. AB - The effects of 17 beta-estradiol, dihydrodydrogesterone, tamoxifen and cyclophosphamide upon parameters of cell maturation (Mucine1 expression), cell proliferation (Cyclin D1 expression) and apoptosis (loss of nuclear DNA) were studied in estrogen receptor positive (ER+) and negative (ER-) human breast cancer cells. Tamoxifen was the most potent inducer of apoptosis in ER+ and ER- breast cancer cells. 17 beta-estradiol in a concentration of 10(-6) M induced proliferation in ER+ cells after 144 h. incubation, while equimolar co-incubation with dihydrodydrogesterone prevented this effect and even induced a significant increase of cell death. It is speculated that the continuous use of combined 17 beta-estradiol plus dihydrodydrogesterone might be given as hormone replacement therapy without increased risk of breast cancer and even may reduce the relapse rate in breast cancer patients. PMID- 12536085 TI - MRI findings in Tolosa-Hunt syndrome before and after systemic corticosteroid therapy. AB - Tolosa-Hunt syndrome (THS) is characterized by painful ophthalmoplegia due to a granulomatous inflammation in the cavernous sinus. Corticosteroid therapy dramatically resolves both the clinical and radiological findings of THS. We present MRI findings of six patients with a clinical history of at least one episode of unilateral or bilateral orbital-periorbital pain, clinical findings of associated paresis of one or more of 3rd, 4th, 5th or 6th cranial nerves. All of the patients revealed an enlargement of the symptomatic cavernous sinus on magnetic resonance imaging (MRI) scans. Five patients revealed total resolution of the clinical findings within 1-8 weeks, following systemic corticosteroid treatment. One patient revealed only minor regression of clinical findings within 2 weeks after the initiation of the treatment, so the cavernous sinus lesion was reevaluated as meningioma on MRI, and the patient underwent surgical resection of the mass with resultant histopathological finding of cavernous sinus meningioma. A follow-up MRI scan was performed for five patients at the end of 8-weeks of steroid therapy. Three of these five patients showed total resolution of the cavernous sinus lesions whereas two of them revealed a partial regression of the cavernous sinus lesions. MRI findings before and after systemic corticosteroid therapy are important diagnostic criteria to put the definitive diagnosis of THS and to differentiate it from other cavernous sinus lesions that simulate THS both clinically and radiologically. PMID- 12536086 TI - Hypoxia-ischemic encephalopathy in full-term neonate: correlation proton MR spectroscopy with MR imaging. AB - INTRODUCTION: To evaluate 1H Magnetic Resonance Spectroscopy (1HMRS) in the diagnosis of hypoxia-ischemic encephalopathy (HIE) of full-term neonates correlated with Magnetic Resonance Imaging (MRI). MATERIALS AND METHODS: Thirty eight cases of full-term neonates diagnosed as HIE clinically were selected to perform MRI and 1HMRS examination. The ages ranged from 7 to 17 days, with median age of 8.2 days. In which, 26 cases were followed up and/or MRI reexamined at 6 months of age or later. Eight healthy neonates, with no evidence of birth asphyxia, also underwent 1HMRS for comparison. SE sequences were used for routine MR examination; point resolved spectroscopy sequence was required for 1HMRS. The metabolites in the spectra includes: N-acetylaspartate (NAA), choline compounds (CHO), creatine compounds (CR), myo-inositol (MI), lactate (LAC), glutamate and glutamine (Glu-Gln). RESULTS: The peaks of NAA were fall in two cases; the peaks of LAC, which were elevated, appeared as typical double-peaks appearance in 26 cases; the peaks of Glu-Gln, which were also elevated, appeared as zigzag appearance in nine cases. The peaks of CR were decreased in 11 cases, while those of MI were increased in seven cases. Mild type of lesions was present on MRI in 12 cases whose LAC/CR ratio lower than 0.5; mild and moderate types of lesions were present in 15 cases whose LAC/CR ratio between 0.5 and 1.5. Whereas, nine cases of severe lesions and two cases of moderate lesions were present on MRI in 11 cases whose LAC/CR ratio greater than 1.5. Twenty-six of 38 cases were followed up and/or MRI reexamined after 6 months, in which, sequelae were present in 12 cases. Among them, eight cases of sequelae in nine cases whose LAC/CR ratio greater than 1.5 were present (account for 88.89%). CONCLUSION: 1HMRS plays an important role to diagnose and predict outcome of HIE. PMID- 12536087 TI - Early-postoperative magnetic resonance imaging in glial tumors: prediction of tumor regrowth and recurrence. AB - OBJECTIVE: This study investigated the value of early-postoperative magnetic resonance (EPMR) imaging in the detection of residual glial tumor and investigated the role of EPMR for the prediction of tumor regrowth and recurrence. METHODS AND MATERIALS: We retrospectively analyzed pre- and post operative magnetic resonance imaging results from 50 adult patients who underwent surgical treatment for supratentorial glial tumor. There were glioblastoma multiforme in 25 patients, astrocytoma (grades II and III) in 11 patients, oligodendroglioma (grades II and III) in 9 patients, and oligoastrocytoma (grades II and III) in 5 patients. EPMR imaging was performed within 24 h after surgery. EPMR findings were compared with the neurosurgeon's intraoperative estimation of gross tumor removal. Patterns of contrast enhancement at the resection site, in residual and developing tumor tissue and blood at the resection site were evaluated on EPMR and in follow-up studies. 'Residual tumor' was defined as contrast enhancing mass at the operative site on EPMR. 'Regrowth' was defined as contrast enhancing mass detected on follow-up in the same location as the primary tumor. 'Recurrence' was defined as appearance of a mass lesion in the brain parenchyma distant from the resection bed during follow-up. RESULTS: Nineteen patients showed no evidence of residual tumor, regrowth, or recurrence on EPMR or any of the later follow-up radiological examinations. EPMR identified 20 cases of residual tumor. Follow-up showed tumor regrowth in 10 patients, and tumor recurrence in 1 case. EPMR showed contrast enhancement of the resection bed in 45 of the 50 patients. Four of the 20 residual tumors showed a thick linear enhancement pattern, and the other 16 cases exhibited thick linear-nodular enhancement. No thin linear enhancement was observed in the residual tumor group. Nine of the 10-regrowth tumors showed a thick linear-nodular enhancement pattern, and one exhibited thin linear enhancement in EPMR. For predicting regrowth tumor EPMR sensitivity was 91%, specificity was 100%, positive predictive value 1; negative predictive value was 0.9375. CONCLUSION: EPMR, depending on the surgical site enhancement pattern, is a valuable means of demonstrating residual tumors, and can be used to predict possible regrowth after surgery. PMID- 12536088 TI - MR features of aneurysmal bone cyst of the mandible and characteristics distinguishing it from other lesions. AB - Aneurysmal bone cyst (ABC) is a relatively rare, non-neoplastic expansile lesion of bone. Bleeding may occur during an operation or biopsy for ABC, as this cyst is an aneurys with numerous pools of blood. Therefore, it is necessary to diagnose ABC before treatment or biopsy. In the present report, we describe the characteristic computed tomography (CT) and magnetic resonance imaging (MRI) features of ABC in the mandible. Based on the literature and on our own experiences, we compare with the features of ABC with the corresponding features of other lesions showing similar conventional radiographic appearances. In the present case, bone-targeting CT showed the characteristic feature, which reflected the histopathological appearance of a partially cystic meshwork divided by coarse septa. MRI showed almost homogeneous intermediate signal intensity, including a partial slight low-signal-intensity area on the T1-weighted image, and homogeneous high signal intensity, which showed a 'bubbly' appearance, on T2 weighted image. On the enhanced T1-weighted image, the intermediate signal intensity areas apart from the areas that showed slight low-signal intensity on the non-enhanced T1-weighted image, were well enhanced. This creates a 'honeycomb' appearance. The 'meshwork' appearance on bone-targeting CT, the 'bubbly' appearance on the T2WI and the 'honeycomb' appearance on Gd-T1WI may be the characteristic features of ABC. PMID- 12536089 TI - Tracheobronchopathia osteochondroplastica presenting as a respiratory insufficiency: diagnosis by bronchoscopy and MRI. AB - Tracheobronchopathia osteochondroplastica (TO) is a rare benign disorder affecting the trachea and occasionally the bronchi. We report a case of TO presenting as a respiratory insufficiency. Chest radiograph revealed an irregular narrowing of the intra thoracic trachea and a parenchymal consolidation of the left lower lobe. Magnetic resonance examination of the chest showed a diffuse irregular thickening of the trachea and central bronchi, which had an intermediate signal intensity with punctiform low signal intensity suggesting calcifications and no contrast enhancement. The diagnosis was confirmed by bronchoscopy and biopsies. PMID- 12536090 TI - Lung parenchyma changes in ankylosing spondylitis: demonstration with high resolution CT and correlation with disease duration. AB - OBJECTIVE: To analyze the spectrum of the lung parenchyma changes in ankylosing spondylitis (AS) with high resolution computed tomography (HRCT) and correlate the findings with disease duration. MATERIAL AND METHODS: Twenty patients (18 male, 2 female) with the diagnosis of AS according to New York criteria were included in the study. None of the patients had history of tuberculosis, prolonged inorganic dust exposure and hospitalization for pneumonia. Seven of the patients were smokers, three patients were ex-smokers, and 10 patients were nonsmokers. The patients were assigned to three groups depending on disease duration. Group 1: patients with disease duration or=6 years but or=11 years (N: 12 patients). HRCT and pulmonary function tests (PFT) were performed in all patients. RESULTS: HRCT demonstrated pathology in 17 patients (85%). Two patients in group 1, 4 patients in group 2 and 11 patients in group 3 had pulmonary parenchyma changes. Emphysema (9/20), septal thickening (9/20) and pleural thickening (9/20) were the most common changes followed by nodule (8/20) and subpleural band formation (7/20). Three patients had apical fibrosis (AF). Septal and pleural thickening (both 4/10) were the most common changes when only nonsmokers were considered. Among nine patients with emphysema three were nonsmokers. CONCLUSION: There is a wide spectrum in pulmonary parenchyma changes in AS. These changes begin in early stages of the disease and increase with disease duration. Although smoking complicates the spectrum of changes in pulmonary parenchyma, they are predominately in the form of interstitial inflammation. PMID- 12536091 TI - Radiological characteristics of pulmonary hydatid disease in children: less common radiological appearances. AB - OBJECTIVE: To evaluate the chest roentgenogram and CT characteristics of pulmonary hydatid disease (PHD). MATERIAL AND METHODS: Forty-seven (27 male and 20 female, aged between 3 and 11 years) consecutive pediatric patients with surgically proven pulmonary hydatid cysts were enrolled for the study. Posteroanterior and lateral chest roentgenograms, CT of the chest, and laboratory findings (latex agglutination, Casoni skin test, and eosinophil count) were obtained from all of the patients. The radiological features (localization, internal architecture, number, diameter) were determined. RESULTS: On CT examination, a total of 79 cysts were determined. On chest roentgenogram, 57 of 79 cysts were detected in all patients. Single cysts were seen in 33 patients, while multiple cysts were seen in 14. Median CT density of the cysts was 21 Hounsfield units (HU) (0-80). There were six giant cysts (>10 cm of cyst diameter). The crescent sign, water lily sign, and air-fluid level were seen in two, five and eight of the cysts, respectively. Apart from the classically described features of pulmonary hydatid cysts of the lung, a crescent-shaped rim of air at the lower end of the cyst (inverse crescent sign) was detected in three cysts. All of the liquid content of the cyst was expelled to the bronchial system (dry cyst sign) was observed in seven cysts. There were two infected cysts. Heavily calcified curvilinear cyst wall was present in one cyst. Pericystic reaction in the lung tissue was observed in five patients. Other features included pleural effusion (n=2), mediastinal shift (n=6) and atelectasis (n=7). CONCLUSIONS: Chest roentgenogram is helpful for diagnosis of intact cysts but, it is impossible to define entire morphology of the complicated cysts. CT imaging recognize certain details of the lesions and discover others that are not visible by conventional radiography. In conclusion, CT examination should be done to elucidate cystic nature of the lung mass and for accurate localization in the preoperative period. In addition, inverse crescent sign should be recognized as feature of pulmonary hydatid cysts on CT. PMID- 12536092 TI - Pulmonary hilar lymph nodes in lung cancer: assessment with 3D-dynamic contrast enhanced MR imaging. AB - PURPOSE: We performed 3D-dynamic MRI on patients with primary lung cancer to identify its usefulness for detecting hilar adenopathy shown at surgery. METHODS AND MATERIALS: 30 consecutive patients with peripheral lung cancer underwent preoperative 3D-dynamic Gd-DTPA-enhanced MRI. Two thoracic radiologists blinded to histopathologic findings reviewed those studies independently for hilar adenopathy visualization. The results were correlated with surgical and histopathologic findings. Interreader agreement for the detection of hilar adenopathy was assessed by means of the kappa statistic. RESULTS: Dynamic MRI demonstrated hilar adenopathy, with or without metastasis revealed at surgery, in all of 15 patients. Adenopathy without metastasis was shown in four patients. Dynamic MRI also revealed metastatic adenopathy in 11 of 12 patients with pathologically proven metastasis. There was only one case with lymph node metastasis that did not have adenopathy either on MRI or even at surgery. The diagnostic accuracy of dynamic MRI for adenopathy with or without metastases revealed at surgery were as follows; sensitivity, 100%; specificity, 100%; positive predictive value, 100%; and negative predictive value, 100%, respectively. The diagnostic accuracy of dynamic MRI for hilar lymph nodes metastasis were as follows; sensitivity, 92%; specificity, 78%; positive predictive value, 73%; and negative predictive value, 93%. Interreader agreement was substantial (kappa=0.73) for detection of hilar adenopathy. CONCLUSION: Hilar adenopathy on 3D-dynamic MRI correlated well with that of surgical finding on patients with primary lung cancer. It may have the potential to make an accurate preoperative evaluation of hilar lymph node metastasis from lung cancer. PMID- 12536093 TI - Comparison of standard reading and computer aided detection (CAD) on a national proficiency test of screening mammography. AB - OBJECTIVE: To evaluate the role of computer aided detection (CAD) in improving the interpretation of screening mammograms MATERIAL AND METHODS: Ten radiologists underwent a proficiency test of screening mammography first by conventional reading and then with the help of CAD. Radiologists were blinded to test results for the whole study duration. Results of conventional and CAD reading were compared in terms of sensitivity and recall rate. Double reading was simulated combining conventional readings of four expert radiologists and compared with CAD reading. RESULTS: Considering all ten readings, cancer was identified in 146 or 153 of 170 cases (85.8 vs. 90.0%; chi(2)=0.99, df=1, P=0.31) and recalls were 106 or 152 of 1330 cases (7.9 vs. 11.4%; chi(2)=8.69, df=1, P=0.003) at conventional or CAD reading, respectively. CAD reading was essentially the same (sensitivity 97.0 vs. 96.0%; chi(2)=7.1, df=1, P=0.93; recall rate 10.7 vs. 10.6%; chi(2)=1.5, df=1, P=0.96) as compared with simulated conventional double reading. CONCLUSION: CAD reading seems to improve the sensitivity of conventional reading while reducing specificity, both effects being of limited size. CAD reading had almost the same performance of simulated conventional double reading, suggesting a possible use of CAD which needs to be confirmed by further studies inclusive of cost-effective analysis. PMID- 12536094 TI - A digital equalisation technique improving visualisation of dense mammary gland and breast periphery in mammography. AB - INTRODUCTION: In mammographic imaging, use of high contrast screen-film combinations results in under-exposed and over-exposed film areas corresponding to dense mammary gland and breast periphery (BP), respectively, characterised by degraded image contrast. A digital equalisation technique was designed and developed in order to deal with the problem of poor visualisation of these regions. METHODS AND MATERIAL: The technique is based on the film-digitiser characteristic curve and a layer model of the breast region, as depicted on a mammogram. It remaps each layer grey level (GL) values by a correction factor that accounts for thickness variation in BP and the presence of dense fibroglandular tissues at the mammary gland. The major steps of the technique are segmentation, to isolate the breast region from mammogram background, and adaptive layer GL remapping. RESULTS: The performance of the technique was initially evaluated on a sample of 60 mammograms. Comparative evaluation between the initial and processed images was performed on the basis of nine anatomical features situated at dense mammary gland and BP. The mammographic images resulting from application of the proposed technique are GL equalised and the visualisation improvement of all anatomical features was found to be statistically significant (P<0.05) or highly significant (P<0.0001). The proposed technique was also compared with contrast-limited adaptive histogram equalisation (CLAHE) and found to be more effective in the visualisation of all anatomical features examined, for both dense breast (DB) and BP. DISCUSSION AND CONCLUSION: Application of the proposed technique results in improved visualisation of both dense mammary gland and BP regions. The proposed technique is independent of breast size, breast symmetry and mammographic view. The technique contributes to breast dose minimisation by eliminating the need for a second acquisition. PMID- 12536095 TI - Hematocolpos in double vagina associated with uterus didelphus: US and MR findings. AB - The malformative syndrome of double vagina in association with uterus didelphus and kidney agenesis is a rare condition, often asymptomatic: if this condition is accompanied by imperforated obstructed hemivagina, the clinical manifestations depend on the presence of hematocolpos. MRI plays an important role for diagnosis because it allows to characterize the nature of the lesion and to evaluate the anatomical details of the uterine malformation. The authors describe this complex genital malformation and discuss the main US and MRI features. PMID- 12536096 TI - A direct comparison of signal behavior between 4.0 and 1.5 T: a phantom study. AB - INTRODUCTION: Higher magnetic fields (>or=3 T) afford higher spatial and/or temporal resolution in MR imaging with contrast agents, however, studies containing direct comparisons of signal intensity among different magnetic fields are substantially sparse. Our aim was to quantify the differences in terms of signal-to-noise ratios (SNRs) and contrast-to-noise ratios (CNRs) between higher and lower ( 5 min) per hour were compared between different periods (total, diurnal, and nocturnal). RESULTS: For the reflux index, the sensibility, the specificity, the positive predictive value and the negative predictive value of the awake period in comparison with the continuous 24 h record, considering the same standards for both periods, were 97%, 46%, 72% and 92% respectively. The ROC graph analysis showed that the cut off adaptation for the reflux index and for the number of reflux per hour, did not allow to improve the day pH-metry performance, since sensibility and specificity moved respectively from 66% to 56% for reflux index, and from 75% to 56% for the number of reflux per hour. CONCLUSION: In children explored for recurrent respiratory diseases, the night record has a bad diagnostic value and a negative diurnal record is strongly predictive of absence of pathological gastroesophageal reflux in these children. Because of the absence of paediatric specific standards for this disorder, it is not possible to reduce the recording time to the diurnal period. PMID- 12536105 TI - [Factors impacting on antiretroviral therapy compliance in HIV positive adolescents]. AB - Is poor treatment observance frequently observed in HIV positive adolescent population, due to the adolescent developmental process, their experience of living with HIV, or lack of information on the disease or treatment? METHODS: We interviewed the 29 HIV positive adolescents followed up in one pediatric reference center. Confidential interviews were performed according to a standard questionnaire by psychologist and research nurse. RESULTS: Seventy-nine percent had stopped at least once a time their treatment who was the major barrier to their sense of freedom behind their adolescence (one-third of them decided to stop it more than one month), although 75% knew the potential consequences and had a good information about their seeks. Fifty-five percent expressed feelings of loneliness or depression, yet 75% consider they had a pleasant life because they had a normal life. In fact, the secret and silence about HIV were the price for this normality. CONCLUSION: Being dependent upon a treatment is a major constraint on the lives of HIV positive adolescents. Although they are fully informed, the deliberate interruption of treatment could attest of their expressed need for autonomy and medical prescription have to be careful with this problem. PMID- 12536106 TI - [Sudden unexpected death in an infant due to a ruptured coronary aneurysm revealing Kawasaki disease]. AB - The early mortality of Kawasaki disease is low, resulting from coronary complications, mainly aneurismal thrombosis with myocardial infarction. The aneurysmal rupture is an exceptional cause of death. CASE REPORT: We report on a six-month-old girl who died suddenly and unexpectedly. The autopsy showed a cardiac tamponade caused by an important ruptured aneurysm of the left coronary artery. Multiple aneurysms, with or without thrombosis, on the right coronary were also present. There was no ischaemic lesion. Three weeks before death, this infant demonstrated several clinical signs of Kawasaki disease whose diagnosis had not been done. CONCLUSION: Even if the rupture of coronary aneurysm is an exceptional initial complication of Kawasaki disease, a better knowledge of the atypical or incomplete forms, particularly below the age of one year, should allow an early diagnosis and treatment, decreasing the risks of coronary complications. This observation furthermore illustrates the interest of post mortem examination in all sudden or unexpected infant deaths. PMID- 12536107 TI - [Chronic recurrent multifocal osteomyelitis of the mandible. A case report]. AB - Chronic recurrent multifocal osteomyelitis (CRMO) is a disorder rarely localized to the lower jaw. CASE REPORT: A fourteen-year-old boy complained of a swollen of his lower jaw. After a CT Scan, a bone biopsy was performed and yielded S. oralis against which an adapted intravenous antibiotherapy was administered without efficacy. The absence of malignant process and the revelation of an other focus of fixation at the Tc bone scan localized on humerus called to mind the diagnosis of CRMO. CONCLUSION: The diagnosis of this disease is difficult and based on a number of concording arguments:clinical and radiological signs of osteomyelitis, multifocal presentation, recurrent relapses and remissions, inaction of antibiotics, elimination of the other differential diagnosis, in particular the infectious osteitis. PMID- 12536108 TI - [Andersen syndrome, ventricular arrhythmias and channelopathy (a case report)]. AB - INTRODUCTION: Recent advances in molecular genetic research have provided new insights into severe ventricular arrhythmias related to channelopathies. CASE REPORT: A case of Andersen's syndrome followed during fourteen years is reported. This rare familial periodic paralysis is characterized by its association with dysmorphic features (micrognatia) and ventricular arrhythmias. COMMENTS: Andersen's syndrome has been attributed to a mutation in the KCNJ2 gene which is involved not only in stabilizing cardiac rhythm, but also in modulating the excitability of skeletal muscle and in morphogenesis. This disease must be distinguished from hyperkalemic periodic paralysis due to a mutation in the skeletal muscle sodium channel gene (SCN4A) and from hypokalemic periodic paralysis related to dihydropyridine receptor mutation (CACNL1A3). Furthermore, it may not be confused with others rhythmic channelopathies (long QT syndromes, catecholaminergic polymorphic ventricular tachycardia and Brugada's syndrome). PMID- 12536109 TI - [Life-threatening Plasmodium falciparum parasitemia, in an infant returning from the tropics]. AB - INTRODUCTION: Severe falciparum malaria is a polymorphous life-threatening disease. Hyperparasitemia is rare among non-immune children returning from tropical areas. CARE REPORT: We report a case of an infant returning from Ivory Coast who presented with fever, prostration and marked thrombocytopenia (22,000 platelets/mm(3)). Blood slide showed Plasmodium falciparum ring forms. Parasite density dramatically increased from 1.7% to 45%, and the child developed several features of severe malaria, according to WHO definition. CONCLUSION: In this non immune infant, prostration and thrombopocytopenia seemed to be earlier predictors of severity than hyperparasitemia and other WHO criteria, which have not been yet validated in travelling children. PMID- 12536110 TI - [Difficult asthma in children]. AB - Difficult asthma in children is defined as the persistence of exacerbations or frequent symptoms requiring rescue bronchodilator, or persistent airway obstruction in spite of treatment with inhaled steroid >/= 800 microg/d beclomethasone or equivalent and beta-2 long acting agonist. Management of difficult asthma in children first requires to identify conditions that may mimic asthma, asthma with bad compliance to treatment, and difficult asthma in relation with avoidable factors that worsen symptoms. The pathological bases of genuine difficult asthma remain unknown. Different patterns have been described according to the cells that are involved (eosinophil, neutrophil), the degree of airway remodeling, or the distal localization of the lesions. Difficult asthma requires specialized management including airway inflammation evaluation. Studies on bronchoalveolar lavage and bronchial mucosa biopsies will perhaps help to better understand the pathophysiology and to improve the management. PMID- 12536111 TI - [The neonatal behavioral observation: a pertinent source of medical informations]. AB - The neonatal behavioral observation is an important source of medical information in three domains: 1) assessment of development which can be done with the Assessment of Preterm Infant Behavior, 2) assessment of pain with the analysis of facial expression using validated pain scales such as the Neonatal Facial Coding System, 3) assessment of brain injuries with the Quality Assessment of General Movements. Such a behavioral observation of the newborn using validated tools is a useful complement of the neuro-imaging techniques. PMID- 12536112 TI - [Management of central precocious puberty]. PMID- 12536113 TI - [Buried penis: a poorly known entity]. PMID- 12536114 TI - [A child dies in hospital. How to help parents bereave?]. PMID- 12536115 TI - [Oral administration of medicines to infants: the pacifier that eases the burden]. PMID- 12536116 TI - [Acute suppurative parotitis in an infant]. PMID- 12536117 TI - [Streptococcal blistering dactylitis]. PMID- 12536118 TI - Inhibition of factor VIIa/tissue factor with nematode anticoagulant protein c2: from unique mechanism to a promising new clinical anticoagulant. AB - Recombinant nematode anticoagulant protein c2 (rNAPc2) is a potent (K(i) = 10 pM) inhibitor of the factor VIIa/tissue factor complex (fVIIa/TF) that involves the pre-requisite binding to either zymogen or activated factor X (fX) prior to the formation of the final quaternary complex with fVIIa/TF. The formation of the binary complex with circulating fX governs the pharmacokinetic profile of rNAPc2 in humans, resulting in a prolonged elimination half-life of >50 h. The clinical antithrombotic potential of rNAPc2 has been evaluated in a phase-II trial in which the incidence of deep-vein thrombosis was reduced over 50% compared to historic controls with low-molecular-weight heparin in patients undergoing knee replacement surgery. A second phase-IIa trial demonstrated the safety of rNAPc2 and the significant suppression of thrombin generation in patients undergoing elective percutaneous coronary intervention treated with standard anticoagulant and antiplatelet therapies. Overall, rNAPc2 is a unique inhibitor of the fVIIa/TF complex and a promising new clinical anticoagulant. PMID- 12536119 TI - Heparin activates antithrombin anticoagulant function by generating new interaction sites (exosites) for blood clotting proteinases. AB - The plasma protein, antithrombin, and its polysaccharide activator, heparin, are essential anticoagulant regulators of blood clotting proteinases that are critical for maintaining hemostasis. Heparin activates antithrombin both by inducing conformational changes in the protein that specifically enhances factor Xa binding and by providing a surface to promote thrombin or factor Xa binding alongside antithrombin in a ternary bridging complex. Although x-ray structures of antithrombin, free and complexed with heparin, have suggested that exposure of a reactive proteinase binding loop is a key feature of conformational activation, mutagenesis of reactive loop residues indicates that the function of this structural change is not to present an optimal loop sequence to target clotting proteinases. Rather, the reactive loop sequence provides only the minimal requirements for recognition by either thrombin or factor Xa, and heparin activation enhances antithrombin recognition by these proteinases through the presentation of exosites outside of the reactive loop. These and other findings suggest that the reactive loop sequence of antithrombin was designed not for optimal recognition by procoagulant proteinases but rather to prevent recognition by the anticoagulant proteinase, activated protein C, thus ensuring that antithrombin functions as an effective anticoagulant. PMID- 12536120 TI - Desmin cytoskeleton: a potential regulator of muscle mitochondrial behavior and function. AB - Alterations in mitochondrial distribution and morphology are associated with normal cellular processes such as cell division and differentiation, as well as a variety of pathological conditions, including muscular dystrophy and cardiomyopathy. These observations have illuminated the necessity for a cellular machinery that mediates mitochondrial behavior and function. One important candidate member of this machinery is the cytoskeleton, all three members of which seem to associate with mitochondria. The role and significance of such association with the intermediate filament (IF) cytoskeleton in muscle was until recently completely unknown. Recent studies with desmin-deficient mice revealed the importance of desmin IFs in mitochondrial behavior and function. This review summarizes recent findings that link desmin cytoskeleton to muscle mitochondrial distribution and function. In particular, hypotheses are presented on the potential mechanism by which desmin's absence from cardiac muscle leads to abnormal mitochondrial behavior and compromised function, potentially responsible for the development of dilated cardiomyopathy and heart failure in desmin-null mice. PMID- 12536121 TI - Functional consequences of mutations in the myosin heavy chain at sites implicated in familial hypertrophic cardiomyopathy. AB - The primary cause of familial hypertrophic cardiomyopathy (FHC) has been attributed to mutations in the genes that encode the contractile proteins of the muscle cell. A majority of these mutations have been found in myosin, the principal component of the thick filament. Most in vitro studies have concluded that FHC mutations cause a loss of function in the biochemical and mechanical properties of myosin. Hypertrophy would then follow as a compensatory mechanism to raise the work and power output of the failing heart. Several recent studies, however, have thrown this mechanism into doubt by providing evidence that FHC mutations in the myosin heavy chain (MHC) can enhance the functional properties of myosin. This review discusses the problems encountered in reaching a definitive answer on the effect of MHC mutations. PMID- 12536122 TI - Addressing the insulin resistance syndrome: a role for the thiazolidinediones. AB - The global epidemic of type 2 diabetes and cardiovascular disease (CVD) is mirrored by increasing prevalence of the Insulin Resistance Syndrome (IRS) or Metabolic Syndrome. Accumulating data indicate that insulin resistance is the common denominator underlying this cluster of related CVD risk factors. Therapeutic interventions that address insulin resistance and other components of the IRS may be of benefit in reducing the significant health and socioeconomic burden presented by diabetes and CVD. Evidence is discussed that the thiazolidinediones, which improve glycemic control by directly targeting insulin resistance, have the additional benefit of improving many of the CVD risk factors in the IRS, and thus have the potential to reduce CVD in patients with type 2 diabetes. PMID- 12536123 TI - Harnessing the response to tissue hypoxia: HIF-1 alpha and therapeutic angiogenesis. AB - In most cell types a family of transcriptional regulatory proteins termed hypoxia inducible factors, of which HIF-1 is the most prevalent, mediates the physiologic response to hypoxia. Although much has been learned over the past decade since the discovery of the first HIF family member, the proximal signaling events linking a decline in oxygen concentration to the activation of HIF-dependent signaling are only now being clarified. Activation of HIF-1 in eukaryotes induces expression of many genes that assist in adapting the organism to an environment in which oxygen is limiting, such as of genes involved in new blood vessel formation, including isoforms of vascular endothelial growth factor and angiopoietins, among others. Targeted expression of constitutively active HIF transgenes to ischemic tissues may be beneficial as a form of therapeutic angiogenesis. PMID- 12536124 TI - CAMP and Ca2+ interplay: a matter of oscillation patterns. AB - Ca(2+) and cAMP signalling pathways are tightly interconnected and the cellular effects mediated by the two second messengers depend strictly on reciprocal modulation. The signalling network that derives from such interplay holds the potential for expanding the information content of the signal enormously and, thus, can contribute substantially to the specificity and diversity of the response. Recent work points to interdependent Ca(2+) and cAMP oscillation patterns as a new paradigm for signal transduction. PMID- 12536125 TI - Na+ channel Nav1.9: in search of a gating mechanism. AB - Voltage-gated Na(+) channels play key roles in generating and propagating action potentials. Their gating is believed to rely exclusively on changes in membrane potential. However, recent data from Blum, Kafitz and Konnerth provide direct evidence that the opening of Na(v)1.9, a member of the voltage-gated Na(+) channel family, is mediated by ligand binding rather than by voltage. This is arguably one of the most influential ideas brought to us in the history of the Na(+) channel field. PMID- 12536126 TI - Towards cracking the code: LIM protein complexes in the spinal cord. AB - Combinatorial transcription codes are used widely in the developing nervous system to specify the development of many distinct cell types. Although this strategy maximizes the use of small numbers of proteins, the molecular basis for imposing specificity remains unclear. A recent study has addressed this question by demonstrating that the favoring of hexameric complexes of specific LIM homeodomain proteins over tetramers could dictate the choice between motor neuron versus interneuron fate. PMID- 12536127 TI - Multiple paths towards repair in multiple sclerosis. AB - Multiple sclerosis (MS) is an inflammatory demyelinating disease of the CNS, affecting approximately 1/1000 individuals in the Western world. Available treatments limit CNS inflammation and strategies to repair damage in the CNS offer the potential of recovery of both tissue and function. With further fundamental knowledge developing, this area is ripe for 'translation' to clinical application. PMID- 12536128 TI - From synchrony to sparseness. AB - The neurons in the antennal lobe of the locust had been shown to encode the identity of odorants using spatially distributed synchronized patterns of neural activity. Recent work describes how such neural patterns are detected. By using non-linear membrane properties, one set of target neurons, the Kenyon cells of the mushroom bodies, are able to act as coincidence detectors, sensitive to synchronized activity. In addition, the specific circuitry between the antennal lobe and the mushroom body refines the spatial-temporal selectivity of the Kenyon cells. In this process, the neural representation of odor identity is transformed from a dense spatial-temporal code into a sparse code. PMID- 12536129 TI - Memory traces unbound. AB - The idea that new memories are initially 'labile' and sensitive to disruption before becoming permanently stored in the wiring of the brain has been dogma for >100 years. Recently, we have revisited the hypothesis that reactivation of a consolidated memory can return it to a labile, sensitive state - in which it can be modified, strengthened, changed or even erased! The data generated from some of the best-described paradigms in memory research, in conjunction with powerful neurobiological technologies, have provided striking support for a very dynamic neurobiological basis of memory, which is beginning to overturn the old dogma. PMID- 12536130 TI - Traffic at the intersection of neurotrophic factor signaling and neurodegeneration. AB - Advances in understanding the biology of neurotrophic factors and their signaling pathways have provided important insights into the normal growth, differentiation and maintenance of neurons. Stimulated by neuropathological observations and genetic discoveries, studies in cell and animal models of neurodegenerative disorders have begun to clarify pathogenetic mechanisms. We examine the intersection of these research themes and identify several potential mechanisms for linking failed neurotrophic factor signaling to neurodegeneration. Studies of nerve growth factor signaling in a mouse model of Down syndrome encourage the views that neuronal dysfunction and atrophy might be linked to failed neurotrophic support and that additional studies focused on this possibility would enhance our understanding of the mechanisms of neurodegenerative disorders and their treatment. PMID- 12536131 TI - The Yin and Yang of NMDA receptor signalling. AB - Ca(2+) entry through the NMDA subtype of glutamate receptors has the power to determine whether neurons survive or die. Too much NMDA receptor activity is harmful to neurons - but so is too little. Is it a case of too much or too little Ca(2+) influx causing cell death or do other factors, such as receptor location or receptor-associated proteins, play a role? Understanding the mechanisms behind this dichotomous signalling is an important area of molecular neuroscience with direct clinical implications. PMID- 12536132 TI - Bridging the gap between genes and behavior: recent advances in the electrophysiological analysis of neural function in Caenorhabditis elegans. AB - The nematode Caenorhabditis elegans has long been popular with researchers interested in fundamental issues of neural development, sensory processing and behavior. Recently, advances in applying electrophysiological techniques to C. elegans have made this genetically tractable organism considerably more attractive to neurobiologists studying the molecular mechanisms of synaptic organization and function. The development of techniques that involve voltage clamp of specific neurons and muscles has allowed the coupling of genetic perturbation techniques with electrophysiological analyses of nervous system function. Recent studies combining these biophysical and genetic techniques have provided novel insights into the mechanisms of presynaptic neurotransmitter release, postsynaptic responses to neurotransmitters and information processing by neural circuits. PMID- 12536133 TI - The neuroanatomical and functional organization of speech perception. AB - A striking property of speech perception is its resilience in the face of acoustic variability (among speech sounds produced by different speakers at different times, for example). The robustness of speech perception might, in part, result from multiple, complementary representations of the input, which operate in both acoustic-phonetic feature-based and articulatory-gestural domains. Recent studies of the anatomical and functional organization of the non human primate auditory cortical system point to multiple, parallel, hierarchically organized processing pathways that involve the temporal, parietal and frontal cortices. Functional neuroimaging evidence indicates that a similar organization might underlie speech perception in humans. These parallel, hierarchical processing 'streams', both within and across hemispheres, might operate on distinguishable, complementary types of representations and subserve complementary types of processing. Two long-opposing views of speech perception have posited a basis either in acoustic feature processing or in gestural motor processing; the view put forward here might help reconcile these positions. PMID- 12536134 TI - How accurate is magnetic resonance imaging of brain function? AB - Since it was introduced a decade ago, functional magnetic resonance imaging (fMRI) has come to dominate research on the human brain. However, fMRI maps are based on secondary metabolic and hemodynamic events that follow neuronal activity, and not on the electrical activity itself. Therefore, the representation provided by fMRI cannot be assumed a priori to be exact. The accuracy of these maps depends on the spatial extent of the metabolic and hemodynamic changes induced by neuronal activity, and the role played by the vasculature in converting these changes to signals detected by magnetic resonance imaging. Significant progress has been made in both areas, suggesting that it is possible to obtain both spatially accurate and quantitative data on brain function from magnetic resonance methodologies. PMID- 12536135 TI - A gain-control model relating nulling results to the duration of dynamic motion aftereffects. AB - Strength of the motion aftereffect (MAE) is most often quantified by its duration, a high-variance and rather 'subjective' measure. With the help of an automatic gain-control model we quantitatively relate nulling-thresholds, adaptation strength, direction discrimination threshold, and duration of the dynamic MAE (dMAE). This shows how the nulling threshold, a more objective two alternative forced-choice measure, relates to the same system property as MAE durations. Two psychophysical experiments to test the model use moving random pixel-arrays with an adjustable luminance signal-to-noise ratio. We measure MAE duration as a function of adaptation strength and compare the results to the model prediction. We then do the same for nulling-thresholds. Model predictions are strongly supported by the psychophysical findings. In a third experiment we test formulae coupling nulling threshold, MAE-duration, and direction discrimination thresholds, by measuring these quantities as a function of speed. For the medium-to-high speed range of these experiments we found that nulling thresholds increase and dMAE-durations decrease about linearly, whereas direction discrimination thresholds increase exponentially with speed. The model description then suggests that the motion-gain decreases, while the noise-gain and model's threshold increase with speed. PMID- 12536136 TI - Adaptation to vertical disparity induced-depth: implications for disparity processing. AB - Depth aftereffects produced by prolonged inspection of an object in depth can be mediated by monocular and binocular depth cues. The adapting mechanisms responsible for such effects have not yet been fully determined. Theories of binocular depth aftereffects typically posit a role of an adaptive horizontal disparity sensitive mechanism, implying multiple cue-specific mechanisms for depth aftereffects. Here we examined whether binocular depth aftereffects can be attributed to such a cue-specific mechanism. In Experiment 1 we did so using a technique allowing us to maintain horizontal disparities and vergence constant for our adaptation stimuli, whilst manipulating simulated depth by virtue of a vertical disparity induced-depth effect. We found that depth aftereffects were almost identical to those produced by adaptation to stimuli of equivalent depth produced by conventional horizontal disparity modulations. In Experiment 2, we examined depth aftereffects following adaptation to apparently frontal surfaces produced by different combinations of horizontal and vertical disparity modulations. Aftereffects were close to zero. These results suggest that binocular depth aftereffects are not due to adaptation of a horizontal disparity sensitive mechanism, and we argue that adaptation occurs at the level of a 3D shape sensitive mechanism derived from multiple cues. Experiment 3 was a control to examine whether the two types of adaptation stimuli in Experiment 1 were indeed perceptually the same, since in theory they may differ if vertical disparities influenced metric depth scaling. We found no evidence of this, and concluded that the two classes of stimuli used in Experiment 1, though consisting of very different patterns of disparity, were perceptually equivalent. PMID- 12536137 TI - Large capacity storage of integrated objects before change blindness. AB - Normal people have a strikingly low ability to detect changes in a visual scene. This has been taken as evidence that the brain represents only a few objects at a time, namely those currently in the focus of attention. In the present study, subjects were asked to detect changes in the orientation of rectangular figures in a textured display across a 1600 ms gray interval. In the first experiment, change detection improved when the location of a possible change was cued during the interval. The cue remained effective during the entire interval, but after the interval, it was ineffective, suggesting that an initially large representation was overwritten by the post-change display. To control for an effect of light intensity during the interval on the decay of the representation, we compared performance with a gray or a white interval screen in a second experiment. We found no difference between these conditions. In the third experiment, attention was occasionally misdirected during the interval by first cueing the wrong figure, before cueing the correct figure. This did not compromise performance compared to a single cue, indicating that when an item is attentionally selected, the representation of yet unchosen items remains available. In the fourth experiment, the cue was shown to be effective when changes in figure size and orientation were randomly mixed. At the time the cue appeared, subjects could not know whether size or orientation would change, therefore these results suggest that the representation contains features in their 'bound' state. Together, these findings indicate that change blindness involves overwriting of a large capacity representation by the post-change display. PMID- 12536138 TI - Utility of a dynamic termination criterion in the ZEST adaptive threshold method. AB - Bayesian adaptive threshold procedures may be run for a fixed number of trials, or may be stopped when the calculated confidence interval for the threshold reaches a selected limit (a dynamic termination criterion). This study used Monte Carlo simulations to determine whether the confidence interval is a useful predictor of errors in the estimated threshold. No difference was found between the distribution of errors in a fixed trial procedure versus a dynamically terminated procedure of the same average number of trials. In addition, the width of the confidence interval failed to usefully predict observer variability arising from a shallow psychometric function slope or increased false positive response probabilities. This study suggests that dynamic termination criteria are of little use in Bayesian adaptive threshold procedures. PMID- 12536139 TI - Segregation from direction differences in dynamic random-dot stimuli. AB - Previous research has shown that a field of random dots in which each dot alternates between a slow and a fast speed, can give rise to the percept of two superimposed sheets of moving dots when the alternations are out of phase or asynchronous with each other [Vis. Res. 35 (1995) 1691]. Under those conditions, observers can discriminate changes in the slow speed independent of changes in the fast speed. The present study investigated whether such motion-based segregation could result when dots alternated between two different directions. Three observers viewed a variety of displays containing two directions of motion, one upward and one oblique, with the task of discriminating small trial-to-trial changes in the direction of the upward component. The oblique direction component also changed direction from trial-to-trial. The field of dots either alternated synchronously (all dots moved in the same direction and switched to the other direction simultaneously) or asynchronously. Results showed that when the dots alternated synchronously between the directions, observers' direction discrimination performance was generally poor. However, when dots switched directions asynchronously, direction discrimination was only slightly elevated in comparison to that produced by a field of dots all moving in a single direction. Additional experiments demonstrated that this performance was not due to judging the global direction of the random-dot display. Thus the visual system had to segregate the stimulus into its component directions before integrating to arrive at the motion signal to be discriminated. It is concluded that for displays comprising elements that alternate between different directions, local direction signals can be used by the human visual system to effectively segregate a display so long as both direction signals are present simultaneously. PMID- 12536140 TI - Stereopsis is perturbed by vergence error. AB - Fixation disparity or vergence noise produce instantaneous vergence errors. These errors are analogous to the imposition of a pedestal disparity, which is known to elevate stereothresholds. In this study, stereothresholds were measured as a function of induced vergence errors in subjects with normal binocular vision. Stereo half-images were viewed in the dark through a custom mirror haploscope. Vergence constant error and vergence variability were induced by introducing horizontal disconjugate position offsets in a pair of moveable mirrors within the haploscope, resulting in forced vergence demands of 6(Delta) base-in to 12(Delta) base-out. In addition, vergence variability was simulated by producing oscillatory disconjugate retinal image motion via motion of the moveable mirrors. The motion of the mirrors was either sinusoidal (frequency=2-6 Hz) or random, with peak-to-peak amplitudes of 0 degrees -0.5 degrees per eye. Stereopsis worsened systematically with induced or simulated VV that exceeded approximately 1.5' disparity. The results were similar regardless of whether the vergence error was induced by forced vergence or was simulated by periodic or random disconjugate retinal image motion. Stereothresholds were invariant with the frequency of disconjugate oscillation, within the range of frequencies and amplitudes tested. Hence, the simulated vergence velocity is not the essential factor that limits stereopsis within Panum's fusional area. The results indicate that the stereothreshold is elevated if the vergence error exceeds a critical value. PMID- 12536141 TI - Stereothresholds with simulated vergence variability and constant error. AB - Stereothresholds are elevated by vergence constant error (fixation disparity), vergence noise, or both. This study investigated the separate and combined effects of simulated vergence constant error and variability on stereothresholds in four normal observers. Targets were 30 arc min bright vertical lines presented separately to the two eyes for 150 ms in darkness. Vergence constant error, simulated as a pedestal disparity, was induced by altering the screen positions of the stereo half-images relative to a previously visible binocular fixation point. Vergence variability was simulated simultaneously by disconjugate motion (amplitude=0-0.5 deg per eye; frequency=2 or 4 Hz) of a pair of scanning mirrors in a Wheatstone stereoscope that was used to present the images to each eye. Various combinations of pedestal disparity and simulated vergence variability produce equivalent stereothresholds whenever the calculated mean deviation (sigma/instantaneous vergence errors//N) of the stimulus from the fixation plane is the same. In particular, stereothresholds are optimal for mean deviations up to approximately 1.4 arc min and then increase according to a power function with an exponent of 0.61. The results imply that vergence constant errors and vergence variability impair stereothresholds because of the resulting mean deviation from the horopter. PMID- 12536142 TI - The selective tuning model of attention: psychophysical evidence for a suppressive annulus around an attended item. AB - The selective tuning model [Artif. Intell. 78 (1995) 507] is a neurobiologically plausible neural network model of visual attention. One of its key predictions is that to simultaneously solve the problems of convergence of neural input and selection of attended items, the portions of the visual neural network that process an attended stimulus must be surrounded by inhibition. To test this hypothesis, we mapped the attentional field around an attended location in a matching task where the subject's attention was directed to a cued target while the distance of a probe item to the target was varied systematically. The main result was that accuracy increased with inter-target separation. The observed pattern of variation of accuracy with distance provided strong evidence in favor of the critical prediction of the model that attention is actively inhibited in the immediate vicinity of an attended location. PMID- 12536143 TI - Cortical OFF-potentials from the S-cone pathway reveal neural damage in early glaucoma. AB - We used background adaptation and silent substitution techniques to record full field ON and OFF-visually evoked cortical potentials (VECPs) selectively from the S-cone and L- and M-cone pathways in 15 patients with primary open angle glaucoma (POAG), and 15 normal controls. The most statistically significant (P=0.01) VECP test for POAG was a voltage change in the S-cone pathway cortical OFF-response. A sensitivity of 93% was achieved, with 60% specificity at minimum error rate. This is presumed to reflect reduced activity in the S-cone and magnocellular visual pathways, and may provide an effective method for research and monitoring change in early glaucoma. PMID- 12536144 TI - Disease-causing mutations in the cellular retinaldehyde binding protein tighten and abolish ligand interactions. AB - Mutations in the human cellular retinaldehyde binding protein (CRALBP) gene cause retinal pathology. To understand the molecular basis of impaired CRALBP function, we have characterized human recombinant CRALBP containing the disease causing mutations R233W or M225K. Protein structures were verified by amino acid analysis and mass spectrometry, retinoid binding properties were evaluated by UV-visible and fluorescence spectroscopy and substrate carrier functions were assayed for recombinant 11-cis-retinol dehydrogenase (rRDH5). The M225K mutant was less soluble than the R233W mutant and lacked retinoid binding capability and substrate carrier function. In contrast, the R233W mutant exhibited solubility comparable to wild type rCRALBP and bound stoichiometric amounts of 11-cis- and 9 cis-retinal with at least 2-fold higher affinity than wild type rCRALBP. Holo R233W significantly decreased the apparent affinity of rRDH5 for 11-cis-retinoid relative to wild type rCRALBP. Analyses by heteronuclear single quantum correlation NMR demonstrated that the R233W protein exhibits a different conformation than wild type rCRALBP, including a different retinoid-binding pocket conformation. The R233W mutant also undergoes less extensive structural changes upon photoisomerization of bound ligand, suggesting a more constrained structure than that of the wild type protein. Overall, the results show that the M225K mutation abolishes and the R233W mutation tightens retinoid binding and both impair CRALBP function in the visual cycle as an 11-cis-retinol acceptor and as a substrate carrier. PMID- 12536145 TI - Identification of a novel domain in two mammalian inositol-polyphosphate 5 phosphatases that mediates membrane ruffle localization. The inositol 5 phosphatase skip localizes to the endoplasmic reticulum and translocates to membrane ruffles following epidermal growth factor stimulation. AB - SKIP (skeletal muscle and kidney enriched inositol phosphatase) is a recently identified phosphatidylinositol 3,4,5-trisphosphate- and phosphatidylinositol 4,5 bisphosphate-specific 5-phosphatase. In this study, we investigated the intracellular localization of SKIP. Indirect immunofluorescence and subcellular fractionation showed that, in serum-starved cells, both endogenous and recombinant SKIP colocalized with markers of the endoplasmic reticulum (ER). Following epidermal growth factor (EGF) stimulation, SKIP transiently translocated to plasma membrane ruffles and colocalized with submembranous actin. Data base searching demonstrated a novel 128-amino acid domain in the C terminus of SKIP, designated SKICH for SKIP carboxyl homology, which is also found in the 107-kDa 5-phosphatase PIPP and in members of the TRAF6-binding protein family. Recombinant SKIP lacking the SKICH domain localized to the ER, but did not translocate to membrane ruffles following EGF stimulation. The SKIP SKICH domain showed perinuclear localization and mediated EGF-stimulated plasma membrane ruffle localization. The SKICH domain of the 5-phosphatase PIPP also mediated plasma membrane ruffle localization. Mutational analysis identified the core sequence within the SKICH domain that mediated constitutive membrane association and C-terminal sequences unique to SKIP that contributed to ER localization. Collectively, these studies demonstrate a novel membrane-targeting domain that serves to recruit SKIP and PIPP to membrane ruffles. PMID- 12536146 TI - Tyrosine dephosphorylation and ethanol inhibition of N-Methyl-D-aspartate receptor function. AB - The inhibitory effect of ethanol on N-methyl-d-aspartate receptors (NMDARs) is well documented in several brain regions. However, the molecular mechanisms by which ethanol affects NMDARs are not well understood. In contrast to the inhibitory effect of ethanol, phosphorylation of the NMDAR potentiates channel currents (Lu, W. Y., Xiong, Z. G., Lei, S., Orser, B. A., Dudek, E., Browning, M. D., and MacDonald, J. F. (1999) Nat. Neurosci. 2, 331-338). We have previously shown that protein kinase C activators induce tyrosine phosphorylation and potentiation of the NMDAR (Grosshans, D. R., Clayton, D. R., Coultrap, S. J., and Browning, M. D. (2002) Nat. Neurosci. 5, 27-33). We therefore hypothesized that the ethanol inhibition of NMDARs might be due to changes in tyrosine phosphorylation of NMDAR subunits. In support of this hypothesis, we found that tyrosine phosphorylation of both NR2A and NR2B subunits was significantly reduced following in situ exposure of hippocampal slices to 100 mm ethanol. Specifically, phosphorylation of tyrosine 1472 on NR2B was reduced 23.5%. These data suggest a possible mechanism by which ethanol may inhibit the NMDAR via activation of a tyrosine phosphatase. Electrophysiological studies demonstrated that ethanol inhibited NMDAR field excitatory postsynaptic potential slope and amplitude to a similar degree as previously reported by our laboratory and others (Schummers, J., Bentz, S., and Browning, M. D. (1997) Alcohol Clin. Exp. Res. 21, 404-408). Inclusion of bpV(phen), a potent phosphotyrosine phosphatase inhibitor, in the recording chamber prior to and during ethanol exposure significantly reduced the inhibitory effect of ethanol on NMDAR field excitatory postsynaptic potentials. Taken together, these data suggest that phosphatase-mediated dephosphorylation of NMDAR subunits may play an important role in mediating the inhibitory effects of ethanol on the N-methyl-D-aspartate receptor. PMID- 12536147 TI - Permanent nucleosome exclusion from the Gal4p-inducible yeast GCY1 promoter. AB - The promoter of the galactose-inducible yeast GCY1 gene allows high rates of basal transcription and is kept free of nucleosomes regardless of growth conditions. The general regulatory factor, Reb1p, as well as the nucleotide sequence of a single Gal4p-binding site, structurally cooperate to exclude nucleosomes from about 480 bp of DNA that spans the UAS(GAL), the Reb1p-binding site, the TATA-box, and the transcriptional initiation sites. Gal4p, which induces transcription of GCY1 about 25-fold in the presence of galactose, is not required for the alteration in chromatin configuration in the promoter upstream region since the hypersensitive site is unchanged when Gal4p is inactive or absent. As soon as either the Reb1p-binding site or the UAS(GAL) or both are mutated, nucleosomes slip into the promoter of GCY1 paralleled by a reduction of basal transcription activity to about 30% in either single mutant and to <10% in the double mutant. In the mutant of the Reb1p-binding site, induction by galactose/Gal4p restores a nucleosome-free state to an extent resembling the GCY1 wild-type promoter, showing that, in principle, activated Gal4p can exclude nucleosomes on its own. Northern blots of GCY1 transcripts confirm that Reb1p modulates basal transcription and has little influence on the galactose-induced state. PMID- 12536148 TI - Tau phosphorylation by cyclin-dependent kinase 5/p39 during brain development reduces its affinity for microtubules. AB - The microtubule-associated protein tau is a developmentally regulated neuronal phosphoprotein. The phosphorylation of tau reduces its ability to bind and stabilize axonal microtubules during axonal growth. Although tau is phosphorylated by cyclin-dependent kinase 5 (Cdk5) in vitro, its in vivo roles remain unclear. Here, we show that tau is phosphorylated by Cdk5/p39 during brain development, resulting in a reduction of its affinity for microtubules. The activity of Cdk5 is tightly regulated by association with its neuronal activators, p35 or p39. The p35 and p39 expression levels were investigated in the developing mouse brain; the p39 expression level was higher in embryonic hind brain and spinal cord and in postnatal cerebral cortex, whereas that of p35 was most prominent in cerebral cortex at earlier stages of development. The ability of Cdk5 to phosphorylate tau was higher when in association with p39 than in association with p35. Tau phosphorylation at Ser-202 and Thr-205 was decreased in Cdk5-/- mouse brain but not in p35-/- mouse brain, suggesting that Cdk5/p39 is responsible for the in vivo phosphorylation of tau at these sites. Our data suggest that tau phosphorylation by Cdk5 may provide the neuronal microtubules with dynamic properties in a region-specific and developmentally regulated manner. PMID- 12536149 TI - Mapping the ligand binding pocket in the cellular retinaldehyde binding protein. AB - Retinoid interactions determine the function of the cellular retinaldehyde binding protein (CRALBP) in the rod visual cycle where it serves as an 11-cis retinol acceptor for the enzymatic isomerization of all-trans- to 11-cis-retinol and as a substrate carrier for 11-cis-retinol dehydrogenase (RDH5). Based on preliminary NMR studies suggesting retinoid interactions with Met and Trp residues, human recombinant CRALBP (rCRALBP) with altered Met or Trp were produced and analyzed for ligand interactions. The primary structures of the purified proteins were verified for mutants M208A, M222A, M225A, W165F, and W244F, then retinoid binding properties and substrate carrier functions were evaluated. All the mutant proteins bound 11-cis- and 9-cis-retinal and therefore were not grossly misfolded. Altered UV-visible spectra and lower retinoid binding affinities were observed for the mutants, supporting modified ligand interactions. Altered kinetic parameters were observed for RDH5 oxidation of 11 cis-retinol bound to rCRALBP mutants M222A, M225A, and W244F, supporting impaired substrate carrier function. Heteronuclear single quantum correlation NMR analyses confirmed localized structural changes upon photoisomerization of rCRALBP-bound 11-cis-retinal and demonstrated ligand-dependent conformational changes for residues Met-208, Met-222, Trp-165, and Trp-244. Furthermore, residues Met-208, Met-222, Met-225, and Trp-244 are within a region exhibiting high homology to the ligand binding cavity of phosphatidylinositol transfer protein. Overall the data implicate Trp-165, Met-208, Met-222, Met-225, and Trp-244 as components of the CRALBP ligand binding cavity. PMID- 12536150 TI - TRPC1 is required for functional store-operated Ca2+ channels. Role of acidic amino acid residues in the S5-S6 region. AB - The exact role of TRPC1 in store-operated calcium influx channel (SOCC) function is not known. We have examined the effect of overexpression of full-length TRPC1, depletion of endogenous TRPC1, and expression of TRPC1 in which the proposed pore region (S5-S6, amino acids (aa) 557-620) was deleted or modified by site-directed mutagenesis on thapsigargin- and carbachol-stimulated SOCC activity in HSG cells. TRPC1 overexpression induced channel activity that was indistinguishable from the endogenous SOCC activity. Transfection with antisense hTRPC1 decreased SOCC activity although characteristics of SOCC-mediated current, I(SOC), were not altered. Expression of TRPC1 Delta 567-793, but not TRPC1 Delta 664-793, induced a similar decrease in SOCC activity. Furthermore, TRPC1 Delta 567-793 was co immunoprecipitated with endogenous TRPC1. Simultaneous substitutions of seven acidic aa in the S5-S6 region (Asp --> Asn and Glu --> Gln) decreased SOCC mediated Ca(2+), but not Na(+), current and induced a left shift in E(rev). Similar effects were induced by E576K or D581K, but not D581N or E615K, substitution. Furthermore, expressed TRPC1 proteins interacted with each other. Together, these data demonstrate that TRPC1 is required for generation of functional SOCC in HSG cells. We suggest that TRPC1 monomers co-assemble to form SOCC and that specific acidic aa residues in the proposed pore region of TRPC1 contribute to Ca(2+) influx. PMID- 12536152 TI - Running a neurogenetic clinic. PMID- 12536153 TI - Basic genetics for the clinical neurologist. PMID- 12536154 TI - Genetic neuromuscular disease. PMID- 12536155 TI - Genetics of movement disorders and ataxia. PMID- 12536156 TI - Surfing for neurogenetics. PMID- 12536157 TI - Genetics of the dementias. PMID- 12536158 TI - Genetics of epilepsy. PMID- 12536160 TI - An early oscillator model: studies on the biodynamics of the piano strike (Bernstein & Popova, 1930). PMID- 12536161 TI - The effects of intermittent vision on prehension under binocular and monocular viewing. AB - The aim of the present study was to examine the effects of intermittent binocular and monocular vision on the preparation and execution of the transport and grasp phases of prehension, and hence the temporal limit of binocular and monocular integration. Participants in two groups (speed or accuracy) performed prehensile movements of two amplitudes (20 and 40 cm) to either a large or small object (6 x 6 x 2 and 6 x 4x 2 cm) under conditions of binocular and monocular viewing. The interval between visual samples was manipulated with liquid crystal goggles (continuous vision, 20on/60off, and 20on/120off ms). A kinematic analysis indicated that participants modified variables associated with the preparation and execution of prehension in the intermittent vision conditions when instructed to emphasize accuracy. Participants instructed to emphasize speed, modified variables associated with the preparation phase only. The impact of intermittent vision was similar under binocular and monocular viewing. Thus, for prehension, it appears that consecutive binocular or monocular samples need to occur less than 60 ms apart in order to be fully integrated for limb control. PMID- 12536162 TI - Prism exposure affects the proprioceptive frames of reference for interlimb rhythmic coordination. AB - Adaptation to prisms can produce a change in felt arm position, termed proprioceptive shift. We studied the effects of prism-induced proprioceptive shift on interlimb rhythmic coordination performed under proprioceptive guidance, in the absence of vision. Relative to interlimb rhythmic coordination performed before prism exposure, the observed steady states of relative phase for postexposure coordination were shifted by a small but reliable amount. The shift was in the direction expected, given the direction of optical displacement. The amount of variability of interlimb rhythmic coordination was unaffected by prism exposure. The results suggest that the same spatial frames of reference altered by prism adaptation are involved in the production of interlimb rhythmic coordination patterns. PMID- 12536163 TI - A role of the basal ganglia in movement: the effect of precues on discrete bi directional movements in Parkinson's disease. AB - The effect of a precue on improving movement initiation (i.e., reaction time; RT) is well understood, whereas its influence on movement execution (i.e., movement time; MT) has rarely been examined. The current study investigated the influence of a directional precue (i.e., left vs. right) on the RT and MT of simple and discrete bi-directional movements in a large sample of Parkinson's disease patients and healthy control participants. Both patients and controls were tested twice, with testing sessions separated by 2 hours. Patients were tested first following an overnight levodopa withdrawal and again after they had taken their medication. Both patients and controls demonstrated a significant RT improvement when information was provided in advance. MT in both healthy participants and medicated patients was, however, slower with the provision of advance information, while unmedicated patients showed no significant MT effects. These results suggest that while the basal ganglia may not be involved in motor program selection, they may dynamically modulate movement execution. PMID- 12536168 TI - Presenilins and Cellular Damage; A Link through Amyloid? PMID- 12536164 TI - Further insights into post-exercise effects on H-reflexes and motor evoked potentials of the flexor carpi radialis muscles. AB - The present study investigated the relative contribution of the cortical and spinal mechanisms for post-exercise excitability changes in human motoneurons. Seven healthy right-handed adults with no known neuromuscular disabilities performed an isometric voluntary wrist flexion at submaximum continuous exertion. After the subjects continued muscle contraction until volitional fatigue, the H reflexes induced by an electric stimulation and motor evoked potentials (MEPs) induced by a transcranial magnetic stimulation (TMS) from a flexor carpi radialis (FCR) muscle were recorded 7 times every 20 s. The H-reflex was used to assess excitability changes at the spinal level, and the MEP was used to study excitability changes at the cortical level. Hreflexes showed a depression (30% of control value) soon after the cessation of wrist flexion and recovered with time thereafter. On the other hand, an early (short latency) MEP showed facilitation immediately after the cessation of wrist flexion (50% of control value) and thereafter decreased. A possible mechanism for the contradictory results of the 2 tests, in spite of focusing on the same motoneuron pool, might be the different test potential sizes between them. In addition, a late (long latency) MEP response appeared with increasing exercise. With regard to the occurrence of late MEP response, a central mechanism may be proposed to explain the origin-that is, neural pathways with a high threshold that do not participate under normal circumstances might respond to an emergency level of muscle exercise, probably reflecting central effects of fatigue. PMID- 12536169 TI - [Treatments of psoriasis: some news]. PMID- 12536170 TI - [Publications of French dermatology professors from 1996 to 1998]. AB - INTRODUCTION: The number and level of the publications reflect the scientific activity of physicians. The assessment of this activity is required for nomination and for the internal promotion of University Professors--Hospital Practitioners (UP-HP). We present the results of such an assessment. MATERIAL AND METHODS: The list of the individual publications of 62 UP-HP was established from research on the Medline on the PubMed website. We eliminated the journals without an Impact Factor and the articles of continued medical training. A score was obtained for each of the authors on each of their publications. This was calculated by multiplying the rank coefficient of the author by the coefficient of the type of publication (letter, clinical case or article) and by the coefficient of the journal (ranging from 1 to 4). The global score of a UP-HP was the sum of the scores per publication. The mean score of each UP-HP was assessed by dividing their global score by the number of their publications. RESULTS: One thousand thirty-one scientific publications in 3 years was divided into 502 articles, 316 clinical cases and 213 letters. The yearly number of publications per UP-HP was a mean of 5.5 (+/- 4.2) ranging from 0 to 17.3. The publications were mainly written in English (p=10(-2)). There were more articles than clinical case reports and letters (p=10(-1)). The yearly number of publications was greatest among those aged 46-55. No difference was noted in gender, function (head of a department or not) or the size of the town. The UP-HP working in a team including other UPs had written a greater number of publications (p=10(-2)). The global scores did not significantly differ between the 3 age ranges considered; conversely, the mean score decreased significantly with age (p=0.04). The UP-HP working in teams including several UPs had significantly higher global score (p=0.03). DISCUSSION: These results show the heterogeneity of the "publishing" activity. The method used in this study for the scientific assessment could be subject to improvement; its principle is indisputable, but its methods of use remain to be specified. The best "publication structure" of young UP-HP may be related to age, demonstrating a decline in scientific activity during the working life, or a cohort-effect--i.e., the progressive improvement in recruitment conditions and scientific activity over the past 20 years. The better performance of the UP-HP working in a team including several UP-HP must be interpreted with care; however, it may help to define a geographical policy of Dermatology. PMID- 12536171 TI - [Malignant blue nevus: clinicopathologic study with AgNOR measurement. Seven cases]. AB - BACKGROUND: Malignant blue nevus is a very rare tumor. Argyrophilic nucleolar organizer regions (AgNORs) have been reported to be both a diagnostic and prognostic clue in various tumors, especially if standardized using an image analysis systems. PATIENTS AND METHODS: Seven cases of malignant blue nevus were retrospectively recorded between 1974 and 1999, and their clinical and pathological features described. Using an image analysis system, AgNOR measurements were studied in all cases. These results were compared with those obtained in 10 cases of commun blue nevus, 10 cases of cellular blue nevus and 10 cases of malignant melanoma. RESULTS: The most frequent location of malignant blue nevus was the scalp. Clinically, the tumor generally consisted of a blue nodule, 2.5 cm in diameter. There was no single histopathological criterion for the diagnosis of malignant blue nevus. AgNOR measurement was significantly higher in malignant blue nevus in comparison with commun blue nevus (p<0.0004) or cellular blue nevus (p<0.012), whereas there was no difference between malignant blue naevus and malignant melanoma (p > 0.50). DISCUSSION: Our results confirm the severe prognosis of malignant blue nevus and highlight the necessity of removing all blue tumours located on the scalp. AgNOR measurement using an image analysis system appears to be a useful tool for the diagnosis of malignant blue nevus, but further studies remain necessary. PMID- 12536172 TI - [Recurrent aphthous stomatitis: treatment with colchicine. An open trial of 54 cases]. AB - INTRODUCTION: Colchicine is a treatment used for aphthous stomatitis but its efficacy has never been proved. We report the results of an open trial of 54 patients treated with colchicine for aphthous stomatitis. The aim of our study was to assess the long and short term efficacy and tolerance of this molecule in this disease. PATIENTS AND METHODS: The case reports of 64 patients presenting with severe, recurrent aphthous stomatitis treated with colchicine between 1986 and 2000 were analyzed retrospectively. Only immunocompetent patients exhibiting idiopathic aphthous stomatitis or Behcet's disease were included. Ten patients were excluded: 4 cases of Behcet's disease treated with colchicine and systemic corticosteroids, 4 cases of aphthous stomatitis secondary to a hematological cause and 2 patients who were lost to follow-up after 3 month's treatment. Colchicine was prescribed at a dose of 1 to 1.5 mg/d for at least 3 months. All the patients were assessed after 3 month's treatment and all were contacted by phone in May 2000. Our end-point criteria were: the frequency and duration of the episodes, intensity of pain and impact on the quality of life of the patients. RESULTS: Fifty-four patients were included: 39 women (mean age=44 +/- 16.8) and 15 men (mean age=49 +/- 13.5). The disease had progressed for a mean of 11.6 +/- 13.5 years. After 3 month's treatment, 12 patients (22 p. 100) no longer had aphta and were in complete remission, 22 patients (41 p. 100) were significantly improved, since the frequency and duration of the lesions had decreased by at least 50 p. 100 and the latter were no longer painful. Treatment failed or tolerance was poor in 20 patients (37 p. 100). Patients were monitored and assessed for a mean of 4.7 years (range: 6 months to 13 years). Six patients were lost to follow-up. Three patients (6 p. 100) were considered as cured, since they were still in complete remission after suspension of colchicine respectively 6, 24 and 72 months later. Fifteen patients (31 p. 100) had still improved. Among the latter, 10 continued colchicine for a mean of 27 months, and 5 patients stopped treatment, which they considered tedious, after a mean of 16 months. These 15 patients noted a significant reduction in the frequency and duration of the lesions. Moreover, the subjective end-point criteria had improved significantly for them not to wish to change treatment. Thirty patients (63 p. 100) were not improved by the treatment. Some benign adverse events were noted in 10 patients (18.5 p. 100) and led to suspension of treatment in 4 cases. DISCUSSION: This study, which is the first in size (n=54) and duration of follow up (4.7 years), showed that colchicine is an efficient preventive treatment of severe aphthous stomatitis for 63 p. 100 of patients after 3 month's treatment and that this improvement was lasting in 37 p. 100 of them. Colchicine should therefore be proposed in first intention in severe recurrent aphthous stomatitis, since it is effective, well tolerated and easy to use. PMID- 12536173 TI - [Positive photobiological investigation in Jessner's lymphocytic infiltration of the skin]. AB - BACKGROUND: Jessner's lymphocytic infiltration of the skin is a rare and benign disorder. Its clinical course is cyclic with remissions and exacerbations. In this disease, photosensitivity was previously noticed by authors and recently demonstrated. We report four new cases with positive photobiological investigation. CASE REPORTS: Four patients (2 males, 2 females), with a mean age 36 years were seen with erythematous papules or discoid plaques on face, arms and upper trunk. The onset of disease occurred after sun exposure in summer. Skin biopsies showed perivascular lymphocytic infiltration in the dermis. All patients relapsed cyclically with incomplete healing during winter. Photobiological investigation elicited skin lesions in a broad spectrum of UV: UVB (2 cases), UVA (1 case), UVA and UVB (1 case). In all patients treatment with oral antimalarials and external photoprotection was effective. DISCUSSION: Our data suggest that photosensitivity history in patients with Jessner's lymphocytic infiltration of the skin should be searched for, and confirmed by provocative phototesting. This relevant event could guide the therapeutic strategy because antimalarials were effective for the Jessner's lymphocytic infiltration cases with photosensitivity. PMID- 12536174 TI - [Anti-TNF alpha in dermatology]. AB - The discovery of the major role of TNF alpha in the physiopathology of certain inflammatory diseases and notably in rheumatoid arthritis and Crohn's disease has led to the development of anti-TNF alpha drugs. These new therapeutic arms issued from bio-technology have rapidly demonstrated their efficacy in the treatment of these two diseases. The anti-TNF alpha arsenal is currently dominated by etanercept, a fusion protein composed of a soluble TNF alpha receptor, and infliximab, a chimeric monoclonal antibody. However, new molecules will soon enrich this arsenal. TNF alpha is a major cytokine of inflammatory diseases of the skin. Many dermatological diseases will probably benefit from these new treatments. Two studies have already demonstrated their interest in cutaneous and articular psoriasis. Encouraging sporadic results suggest other potential indications (Behcet's disease, bullous dermatitis, neutrophilic dermatitis, toxic epidermal necrolysis, systemic vascularitis,.). These promising new treatments, although expensive, and with yet unknown long term side effects, justify rigorous assessment of their efficacy and tolerance in each indication. Here again the dermatologist has a major role to play in post-marketing pharmacovigilance. PMID- 12536175 TI - [Contact dermatitis with Hirudo medicinalis]. AB - BACKGROUND: Hirudine is the first natural anti-coagulant ever described. It is used for its anti-coagulant properties in plastic surgery or for the treatment of post-phlebitic diseases. Natural hirudine is extracted from the saliva of the Hirudo medicinalis leech, but it can also be found in crushed leech and included in a cream (Hirucreme). Side effects to hirudine are considered to be rare. CASE REPORT: We report a contact eczema caused by an extract from the medicinal Hirudo medicinalis leech. This was confirmed by the patch tests. However, we noticed a negativity of these tests with two analogs of the recombinant hirudine. DISCUSSION: Several cases of contact dermatitis with Hirucreme have been described. The analogs of recombinant hirudine, which share similar biological activity, have a very close molecular structure. They are indicated via the systemic route for thrombopenia related to heparin for the prevention of severe thromboses. The negative patch tests does not allow definite conclusion, but they prove that these molecules do not always lead to cross-allergies. PMID- 12536176 TI - [Perforating skin ulcers occurring in an adult with dermatomyositis]. AB - BACKGROUND: Skin ulcerations are rarely reported in dermatomyositis of the adult. We report on a case of perforating ulcers resistant to classical treatments in a woman with dermatomyositis. CASE REPORT: A 34 year-old woman, treated for typical dermatomyositis, developed a few weeks after starting her treatment, multiple perforating skin ulcers on her shoulders, elbows and wrists though general signs had completely disappeared. In spite of the increase in oral steroids, the patient presented again with worsened lesions and a new perforating ulcer on the hand which lead to the breaking of a tendon. She underwent hand surgery and was also treated with a bolus of steroids IV (1 g daily during 3 days), followed by oral steroids, IM methotrexate and antimalarial drugs. The lesions healed completely after one month, with no atrophy. DISCUSSION: Skin ulcers are rarely described in dermatomyosites of the adult and are associated with underlying vasculitis and bad prognosis. Our case is remarkable by the absence of vasculitis, a very slow cure requiring aggressive treatment and excellent prognosis after three years of follow-up. PMID- 12536177 TI - [Juvenile colloid milium]. AB - INTRODUCTION: Colloid Milium is a rare disease. It is characterized by the development, on sun-exposed areas, of clusters of yellow and translucid, hard papules, that seeps a gelatinous substance when opened. This dermatitis predominantly affects elderly patients and, exceptionally, children. We report a case of juvenile colloid milium. OBSERVATION: A 14 year-old boy consulted for papular, translucid or amber-brown lesions, occasionally hemorrhagic on the cheeks, nose upper lip and the upper edge of the helix of both ears. He was born to consanguineous parents. Living in a rural area, he was exposed to sun during agricultural activities. The lesions had progressed since he was 6 years old and were exacerbated in the summertime. There was no similar case in the family. Histological exploration of a papular lesion showed areas of atrophic epidermis and the presence of a few vacuolized keratinocytes. The epidermis was lifted by nodular lumps of amorphous eosinophilic material deposited in the superficial dermis. Staining with Congo red was negative. DISCUSSION: Juvenile colloid milium is a benign but unesthetic dermatitis. The inducing role of sun exposure is obvious, as in colloid milium of adults, but other pathogenic hypotheses also exist. PMID- 12536178 TI - [Ectopic breast cancer of the axilla]. AB - INTRODUCTION: Breast cancer is the most frequent cancer occurring in women. These cancers may rarely develop within ectopic breast tissue. We report a case of ectopic axillary lobular carcinoma presenting as an isolated skin lesion with prolonged evolution. CASE-REPORT: A 62 year-old women had a cutaneous lesion located in her left axilla for the past 4 years. The skin biopsy showed an infiltrating lobular carcinoma. A mammogram and an MRI were as unremarkable as two past mammograms performed since the beginning of the skin lesion. Surgical resection with lymph node dissection, associated with radiotherapy and hormonotherapy with tamoxifen, led to complete remission. DISCUSSION: Ectopic breast tissue is subject to physiological and pathologic changes such as benign or malignant tumours. Cancer occurring in ectopic breast tissue remains rare but this diagnosis must be suspected when confronted with a skin lesion or mass located near the normal breasts. PMID- 12536179 TI - [Genital leiomyomas of the male nipple: two cases]. AB - BACKGROUND: Genital leiomyoma is a rare benign solitary skin tumor, not painful, developed from smooth muscle. Genital leiomyoma arising from the nipple is extremely rare, especially in males. CASES REPORT: A 47 year-old male had a 2 cm cutaneous plaque with nodules located on the right nipple. This plaque was circumscribed, erythematous, pruriginous and was not painful. The lesion had been noticed by the patient two years ago. A 37 year-old male showed a 1.5 cm cutaneous plaque located on the left nipple. The plaque was slightly erythematous, pruriginous, not painful and had been noticed by the patient 5 years earlier. Histology provided the diagnosis of genital leiomyoma in both cases. No surgical therapy was performed. DISCUSSION: Cutaneous leiomyomas are classified in 3 types regarding their origin: multiple or solitary piloleiomyoma, arising from arectores pilorum muscles, solitary genital leiomyoma, arising from the dartoic, vulvar, or mammillary muscles, and solitary angioleiomyoma, arising from the vein muscles. Clinically, genital leiomyoma is a 1 cm diameter solitary erythematous, firm nodule. According to many authors, genital leiomyoma is asymptomatic, but in the 2 patients, the lesions were pruriginous. Surgical excision is usually performed. PMID- 12536180 TI - [The recurrence of syphilis: 3 cases in HIV sero-negative patients]. PMID- 12536181 TI - [Infiltration of the nostril]. PMID- 12536182 TI - [Cutaneous nodules and hematuria]. PMID- 12536183 TI - [Psoriatic arthritis]. PMID- 12536184 TI - [Psoriasis of the scalp]. PMID- 12536185 TI - [Hypersensitivity to corticosteroids of the D1 group]. PMID- 12536186 TI - [Hypersensitivity to budesonide]. PMID- 12536189 TI - [Is still necessary to distinguish seborrheic dermatis and psorasis?]. PMID- 12536190 TI - The genes and proteins associated with poly-cystic kidney diseases. AB - Genetically based polycystic kidney diseases include autosomal dominant (ADPKD) and recessive (ARPKD) polycystic kidney diseases, nephronophthisis and medullary cystic disease. The PKD1 and PKD2 genes responsible for ADPKD and their respective encoded proteins polycystin-1 and polycystin-2 are under intense study and clues are developing as to their function and roles in the disease process. Structure-function analysis suggests that polycystins form multiprotein complexes with focal adhesion and cell-cell adherens junction proteins, which then initiate intracellular signaling events culminating in regulation of transcription of genes controlling proliferation and differentiation. Although less is known about the PKHD-encoded fibrocystin responsible for ARPKD or about the NPH1-encoded nephrocystin responsible for nephronophthisis, it is proposed that they function in the same cellular pathway involving protein-protein interactions, signal transduction and regulation of gene transcription. ADPKD epithelia are more adherent to collagen, less migratory, fail to recruit FAK to polycystin complexes and show aberrant, persistent expression of the fetal genes Erb-B2 and beta2 subunit of NaK-ATPase after birth. It is suggested that the function of the polycystin complex is to act as a key developmental regulator of renal tubule morphogenesis. PMID- 12536191 TI - Simultaneous pancreas-kidney and pancreas transplantation. AB - The best available method currently for achieving steady normoglycemia in individuals with type 1 diabetes mellitus (DM) is replacing the pancreas, e.g. whole pancreas transplantation. Pancreatic transplantation, as either simultaneous pancreas-kidney (SPK) or solitary pancreas transplantation alone (PTA), has moved beyond simple metabolic or quality-of-life goals. It is now an effective treatment to reverse or minimize metabolic abnormalities and complications of type 1 DM as well as potentially extend the life span of those afflicted by type 1 DM and its many co-morbid complications. Candidates for SPK and PTA transplantation need to meet various criteria even to undergo the transplant procedure and receive a pancreatic allograft that is deemed suitable. SPK and PTA recipients, though free from insulin use, still may encounter common post-transplant medical complications, e.g. cardiovascular disease, high blood pressure, as well as complications unique to SPK and PTA transplantation. The advantages of PTA and SPK transplantation are frankly now more obvious as improvements in surgical technique and new immunosuppression have made an increasing number of PTA and SPK transplants viable and functional long-term. The idea of pancreas transplantation can be touted as a therapeutic advance for type 1 DM. It can improve survival and limit many diabetic-related complications, while improving quality of life, especially in those individuals also afflicted with diabetic-related kidney disease. PMID- 12536192 TI - [Predicting factors of vesicoureteral evolution in children]. AB - BACKGROUND: Despite the large number of children with reflux, management among urologists is still controversial. One of the most debated aspects is the choice between observation treatment or surgical treatment. METHODS: We assessed the natural course of children with vesico-ureteral reflux in the period 1990-1995, to correlate factors and identify patients with high risk of renal damage. We retrospectively reviewed the clinical course of 80 children with vesicoureteral reflux. Thirty-two were diagnosed during prenatal ultrasound scan. In the other cases urologic evaluation was requested because of urinary tract infections. The following data were analyzed: medical records, diagnostic and follow-up cystogram, renal imaging, medical therapy or surgical treatment carried out according to the reflux grade, diagnostic age, congenital reflux nephropathy or postnatal acquired scarring, voiding patterns, spontaneous resolution during medical management. Follow up ranged from 5 to 10 years. RESULTS: Vesicoureteral reflux resolved spontaneously in 29 patients: 25 were affected from moderate reflux, 4 from IV grade reflux. Surgical correction was carried out in 32 patients. Endoscopic treatment was performed in 25. Twenty-two children are still receiving prophylaxis and 12 were lost to follow-up. Congenital renal pathology correlate with poor outcome. CONCLUSIONS: The conclusion is drawn that there is a wide clinical variability in children with vescicoureteral reflux. The most important is host's susceptibility to urinary tract infection, but the severity of reflux, age of patients and congenital reflux nephropathy influence prognosis and long-term outcome. PMID- 12536193 TI - [Erectile dysfunction in Peyronie's disease. Experience in 132 patients]. AB - BACKGROUND: The exact percentage of sexual dysfunctions resulting from Peyronie's disease remains very difficult to evaluate even today. The purpose of the present paper is to examine the series observed in our Department in order to make an assessment of the erectile function of patients found to be suffering from Peyronie's disease. METHODS: Over the course of 9 years, 132 patients have come to our observation; they were originally examined for sexual function disturbances or for other pathologies of a uro-andrological nature. RESULTS: In all, a total of 208 lesions were observed. Only 21 patients (22.5%) were found to be completely asymptomatic at the time of diagnosis. The most frequent symptom was shaft curvature (alone or in association with pain: 47%). The most commonly observed site for plaques was the back of the corpus cavernosum (43%). Impotentia erigendi was observed in 29 patients (22%) whereas impotentia coeundi was found in 50 (38). The functionally most significant lesions were those affecting the septum or in some way involved with the septum: 19 of 29 patients with impotentia erigendi presented such lesions. CONCLUSIONS: Considering the generally progressive course of the disease, the therapeutic approach taken by our Department is to fit a prosthesis to patients who have been suffering from Peyronie's disease for at least six months; conservative therapy is only undertaken in the rare cases of patients who have a perfectly preserved erection but who suffer from impotentia coeundi (as a result of curvature or pain). PMID- 12536194 TI - [Microproteinuria as a marker of renal damage in children]. AB - BACKGROUND: Elevated urinary levels of microproteinuria or renal enzyme have shown to be associated with renal injury. Data collected in children with vesico ureteral reflux (VUR) or hydronephrosis have been evaluated to identify a means of predicting renal damage. METHODS: Levels of urinary microproteinuria (N-acetyl beta-glucosaminidase, aminoalanina peptidase, lisozyme, beta(2)-microglobulin) were evaluated with immunoassay in catheterized or voided urine. The levels of urinary creatinine were also determined. Of the 85 children enrolled in this study 22 were affected from reflux, 16 had hydronephrosis. Urine was obtained from the bladder in all children and also from renal pelvis in 9 patients at surgery for ureteropelvic junction obstruction. In addition, urine was obtained from 27 patients with a variety of pathophysiological conditions (neoplasia, ematuria, parenteral nutrition, ecc.). Normal healthy controls were performed in 20 children who volunteered for the study. RESULTS: In the group of children affected from reflux urinary microprotein elevated levels were in 18. In children with hydronephrosis pathological findings were observed in 2 cases. Abnormal pattern was found in 27 patients as response to various agents. Data obtained in healthy children were normal. Sensitivity in reflecting renal damage makes urinary enzymes measurement attractive as an indicator of renal dysfunction. CONCLUSIONS: Urinary microproteinuria evaluation may be a useful marker of progression of renal injury in children with reflux. However, excretion level of these microproteins is not helpful in identifying upper tract obstruction in small children, but values arise in those left untreated. PMID- 12536195 TI - [Chronic renal failure and tuberous sclerosis. Report of two clinical cases]. AB - Tuberous sclerosis complex (TSC) is an autosomal dominant hereditary disease characterized by a high rate of spontaneous mutations involving at least two loci: TSC(1) (9q34) and TSC(2) (16p13). It results in hamartomas or tumours which can affect a variety of organs, most commonly the brain, skin and kidneys. At least half of patients with TSC have underlying renal pathology, most commonly angiomyolipomas (AML) and/or cysts with, more rarely, adenocarcinoma, but oncocytomas, sarcomas, interstitial fibrosis and glomerulosclerosis have all been reported. Renal disorders may be asymptomatic or associated with acute lumbar ache, hematuria, abdominal mass, retroperitoneal hemorrhage. Renal failure is infrequent. The diagnosis of this disease is often performed, as in the present cases, very late and it is made possible by radiological examinations such as TC scan o RMI (when renal failure is present), usually performed after macrohaematuria or abdominal or renal colics or renal failure. When fatty tissue cannot be demonstrated within renal lesion (as in the female case), biopsy can be undertaken to exclude malignancy. Histology at the edge of an AML may look like renal carcinoma, but recent studies suggest that it can be differentiated by staining for HMB-45 which is positive in AML and negative in carcinoma. Two cases of tuberous sclerosis with different neurological fenotype, with bilateral renal angiomyolipomatosis and heavy renal failure, are presented. PMID- 12536196 TI - Ribozyme-mediated cleavage of the human survivin mRNA and inhibition of antiapoptotic function of survivin in MCF-7 cells. AB - Survivin is a new member of the inhibitor of apoptosis protein (IAP) family that is implicated in the control of cell proliferation and the regulation of cell life span. This protein is selectively expressed in most human carcinomas but not in normal adult tissues. To down-regulate a human survivin expression as a strategy for cancer gene therapy, we designed two hammerhead ribozymes (RZ-1, RZ 2) targeting human survivin mRNA. RZ-1 and RZ-2 efficiently cleaved the human survivin mRNA at nucleotide positions +279 and +289, which was identified by in vitro cleavage assay using in vitro transcribed ribozymes and truncated survivin mRNA substrate. To investigate the function of the ribozymes in cells, the sequences of the ribozymes were cloned into replication-deficient adenoviral vector and transferred to breast cancer cell, MCF-7. The infection with adenovirus encoding the ribozymes resulted in a significant reduction of survivin mRNA (74% and 73%, respectively) and protein. As revealed by nuclear condensation/ fragmentation and flow cytometry analysis, inhibition of survivin gene by ribozymes increased apoptosis and sensitivity induced by etoposide or serum starvation. Our results suggest that the designed hammerhead ribozymes against survivin mRNA are good candidates for feasible gene therapy in the treatment of cancer. PMID- 12536197 TI - Antitumor effect of secreted Flt3-ligand can act at distant tumor sites in a murine model of head and neck cancer. AB - The Flt3 ligand (Flt3-L) manifests antitumor activity, presumably due to its capacity to recruit dendritic cells and cause their proliferation. To assess whether local production of Flt3-L can mediate a "distant bystander" effect, murine B4B8 squamous cell carcinoma cells were transfected with a plasmid encoding a secretory form of Flt3-L to produce B4B8FL cells. Similarly, B4B8FL and B4B8 cells were transfected with herpes simplex virus thymidine kinase (HSVTK) to produce B4B8TK and B4B8FL/TK cells, which should be sensitive to ganciclovir (GCV), to know whether the effects of Flt3-L and HSVTK/GCV would be synergistic. To test for a distant bystander effect in vivo, B4B8FL, B4B8TK, and B4B8FL/TK cells were injected subcutaneously into the left flank of syngeneic Balb/c mice, and naive B4B8 cells were injected into the right flank. The formation of tumors derived from B4B8FL and B4B8FL/TK cells was significantly delayed in both flanks compared with naive B4B8 and B4B8TK cells. Growth of B4B8TK tumors in the ipsilateral flank was retarded following GCV treatment, but in contrast to B4B8FL and B4B8FL/TK cells, no distant bystander effect in the contralateral flank was observed. Immunohistochemistry showed lymphocytic infiltrates in both flanks of the B4B8FL and B4B8FL/TK groups. The data indicate that in these cells, local secretion of Flt3-L is sufficient to evoke a distant bystander effect but that expression of HSVTK, even after GCV administration, is not. Furthermore, the combination of local Flt3-L and HSVTK production, together with GCV administration, does not enhance the distant bystander effect produced by Flt3-L alone. PMID- 12536198 TI - Bifidobacterium adolescentis as a delivery system of endostatin for cancer gene therapy: selective inhibitor of angiogenesis and hypoxic tumor growth. AB - In order to overcome difficulties that hampered widespread application of antiangiogenesis in cancer therapy, a highly specific delivery system may be engaged in vivo to deliver and express antiangiogenic genes. We selected a strain of Bifidobacterium adolescentis (B. adolescentis) as the delivery system to transport endostatin gene to solid tumors. B. adolescentis with endostatin gene were injected into tumor-bearing mice through the tail vein. After the mice were sacrificed, the tumor and some normal tissues of the mice were examined. B. adolescentis were only found in the tumors and no bacilli were found in other normal tissues. Also, a strong inhibition of angiogenesis had been shown to inhibit local tumor growth in the administrated group. These results suggested that B. adolescentis only germinated and proliferated in solid tumors and might be a highly specific and efficient vector for transporting anticancer genes into target tumor in cancer gene therapy. PMID- 12536199 TI - Stable expression of antisense urokinase mRNA inhibits the proliferation and invasion of human hepatocellular carcinoma cells. AB - Urokinase-type plasminogen activator (u-PA) plays a key role in malignant tumor behavior. We have previously shown that the expression of high levels of u-PA mRNA in human hepatocellular carcinoma (HCC) biopsies was inversely correlated with the survival of the patients. In order to evaluate the involvement of u-PA in the invasive and infiltrating properties of HCC cells, the SKHep1C3 cell line was stably transfected with an expression vector containing the 5' portion (257 bp) of u-PA cDNA in the antisense orientation. u-PA mRNA expression and its protein level and enzymatic activity were specifically inhibited in the antisense transfectants. A comparable inhibition of the u-PA receptor (u-PAR) mRNA and protein was also evidenced in the antisense transfected cells compared with the control ones. At the functional level, the SKHep1C3-AS cells showed a significant reduction in proliferation, Matrigel invasion, and motility assays compared to parental and vector-alone cells. These results indicate that u-PA is an essential factor in the growth and invasiveness of human hepatocarcinoma cells. Antisense u PA strategy might be a potential approach to reduce tumor growth as well as the invasive capacity of the malignant cells in HCC. PMID- 12536200 TI - Generation of an adenoviral vector containing an addition of a heterologous ligand to the serotype 3 fiber knob. AB - As an initial assessment of the feasibility of employing the adenovirus serotype 3 (Ad3) fiber knob as a locale for introducing a tropism-modifying motif, we generated an adenoviral vector containing a six-histidine tag genetically fused to the carboxy-terminus of the Ad3 fiber knob. The heterologous tag proved to be accessible for binding in the context of the virion and, moreover, had rendered the modified vector capable of mediating gene transfer through an artificial, non Ad3 receptor. PMID- 12536201 TI - RNA interference is a functional pathway with therapeutic potential in human myeloid leukemia cell lines. AB - BACKGROUND: RNA interference (RNAi) is a cellular pathway of gene silencing in a sequence-specific manner at the messenger RNA level. The basic mechanism behind RNAi is the breaking of a double-stranded RNA (dsRNA) matching a specific gene sequence into short pieces called short interfering RNA, which trigger the degradation of mRNA that matches its sequence. In this study, we explored the effects of RNAi in reducing the target gene expression in human myeloid leukemia cell lines. METHODS: Four myeloid leukemia cell lines (HL-60, U937, THP-1, and K562) were transfected with dsRNA duplexes corresponding to the endogenous c-raf and bcl-2 genes and the gene expression inhibition was assessed. The effect of RNAi on cell differentiation was studied; the apoptosis induction and the sensitization of the leukemia cell lines to etoposide and daunorubicin were quantified by flowcytometric methods. RESULTS: Transfection of the myeloid leukemia cell lines with dsRNA corresponding to c-raf and bcl-2 genes decreased the expression of Raf-1 and Bcl-2 proteins. RNAi for c-raf gene blocked the appearance of the monocytic differentiation induced by treatment with TPA. Combined RNAi for c-raf and bcl-2 induced apoptosis in HL-60, U937, and THP-1 cells and increased chemosensitivity to etoposide and daunorubicin. CONCLUSIONS: RNAi is a functional pathway in human myeloid leukemia cell lines and combined RNAi of c-raf and bcl-2 genes may represent a novel approach to leukemia, providing a means to overcome the resistance to chemotherapeutic agents and ultimately to augment the efficacy of chemotherapy in myeloid leukemia. PMID- 12536202 TI - Fas ligand-expressing tumors induce tumor-specific protective immunity in the inoculated hosts but vaccination with the apoptotic tumors suppresses antitumor immunity. AB - The interaction between Fas and Fas ligand (FasL) is involved in the apoptotic death of a number of cells including lymphocytes. Forced expression of FasL in tumors can induce apoptosis of infiltrating Fas-positive T cells; accordingly, tumors can survive in the milieu of systemic immune responses. However, FasL expressing murine lung carcinoma (A11) and melanoma (B16) cells did not develop subcutaneous tumors and FasL-expressing A11 (A11/FasL) cells produced few spontaneous lung metastatic foci in syngeneic mice. The mice that rejected A11/FasL cells were resistant to subsequent challenge of parent A11 but not irrelevant B16 cells. Vaccination of mice with UV-treated A11/FasL, but not UV treated A11 cells, however, augmented the growth rate of A11 but not B16 tumors, both of which were subsequently inoculated. The number of lung metastatic foci of A11 cells was also increased in the mice that received UV-treated A11/FasL but not UV-treated A11 cells. Intraperitoneal injection of UV-treated A11/FasL cells resulted in the production of larger amounts of immunosuppressive TGF-beta in peritoneal exudate than that of UV-treated A11 cells. Expression of the CD80 costimulatory molecule in tissues where UV-treated A11/FasL cells were inoculated was lower than the expression at an untreated A11/FasL-injected site. Our results indicated that apoptotic FasL-expressing tumor cells could impair host immune responses against the tumors, in contrast to potent antitumor immunity generated by viable FasL-expressing tumors. PMID- 12536203 TI - Selective suppression of cathepsin L by antisense cDNA impairs human brain tumor cell invasion in vitro and promotes apoptosis. AB - Invasion and metastasis of certain tumors are accompanied by increased mRNA protein levels and enzymatic activity of cathepsin L. Cathepsin L has also been suggested to play a role in the proteolytic cascades associated with apoptosis. To investigate the role of cathepsin L in brain tumor invasion and apoptosis, the human glioma cell line, IPTP, was stably transfected with full-length antisense and sense cDNA of cathepsin L. Down-regulation of cathepsin L by antisense cDNA significantly impaired (up to 70%) glioma cell invasion in vitro and markedly increased glioma cell apoptosis induced by staurosporine. Compared to control and parental cell lines, antisense down-regulation of cathepsin L was associated with an earlier induction of caspase-3 activity. Up-regulation of cathepsin L activity by sense cDNA was associated with reduced apoptosis and later induction of caspase-3 activity. Moreover, down-regulation of cathepsin L lowered the expression of antiapoptotic protein Bcl-2, whereas up-regulation increased the expression of Bcl-2, indicating that cathepsin L acts upstream of caspase-3. These data show that cathepsin L is an important protein mediating the malignancy of gliomas and its inhibition may diminish their invasion and lead to increased tumor cell apoptosis by reducing apoptotic threshold. PMID- 12536204 TI - Adenovirus E1a protein enhances the cytotoxic effects of the herpes thymidine kinase-ganciclovir system. AB - Cancer gene therapy based on the use of suicide genes, such as the thymidine kinase gene, is not producing satisfactory results. Several approaches have been delineated to enhance the therapeutic responses, including augmentation of the bystander effect, the combination of the herpes simplex virus thymidine kinase ganciclovir (HSVTK-GCV) system into replication competent adenoviruses and others. Moreover, because usually less than 20% of human malignant cells are in S phase, the HSVTK-GCV system is not as efficient as expected. To increase the cytotoxic effects of the HSVTK-GCV system, we hypothesized that concomitant expression of E1a protein, which drives cells to proliferation and S-phase, could increase the effects of the HSVTK-GCV system. Several retroviruses were constructed carrying bicistronic sequences of TK and E1a 12S genes under the control of the CMV promoter. The constructions were tested in murine (NIH-3T3, MSC11A5) and human cells (IMR90, HeLa, MDA-MB435). A clear increase of the HSVTK GCV system killing effect in nonconfluent cells was observed in the cells studied, especially in NIH-3T3, MSC11A5, IMR90, and MDA-MB435 expressing cells. In confluence, the NIH3T3 and IMR90 E1a-TK-expressing cells were also very sensitive and most malignant E1a-TK-expressing cells showed an irreversible G2-M cell cycle arrest. Moreover, the concomitant expression of adenovirus E1a and the HSVTK-GCV system increased the sensitivity to anticancer agents such as cisplatin. These results show that adenovirus E1a protein expression clearly enhances the cytotoxic effects of the HSVTK-GCV system and the response to treatment with cisplatin. PMID- 12536205 TI - Coat protein fold and maturation transition of bacteriophage P22 seen at subnanometer resolutions. AB - Bacteriophage P22 is a prototypical biological machine used for studying protein complex assembly and capsid maturation. Using cryo-EM, we solved the structures of P22 before and after the capsid maturation at 8.5 A and 9.5 A resolutions, respectively. These structures allowed visualization of alpha-helices and beta sheets from which the capsid protein fold is derived. The capsid fold is similar to that of the coat protein of HK97 bacteriophage. The cryo-EM shows that a large conformational change of the P22 capsid during maturation transition involves not only the domain movement of individual subunits, but also refolding of the capsid protein. PMID- 12536206 TI - A dynamic mechanism of nuclear receptor activation and its perturbation in a human disease. AB - Nuclear receptors are transcription factors that activate gene expression in response to ligands. The C-terminal helix (helix 12) of the ligand-binding domain plays a critical role in the activation mechanism. When bound to activating ligands, helix 12 adopts a conformation that promotes the binding of co-activator proteins. Helix 12 also adopts this 'active' position in several ligand-free structures, raising questions as to the exact role of helix 12. We proposed that the dynamic properties of helix 12 may be critical for the activation mechanism and, to test this, have used fluorescence anisotropy techniques to directly monitor the mobility of helix 12 in PPARgamma. Our results suggest that helix 12 is significantly more mobile than the main body of the protein. Upon ligand binding, helix 12 shows reduced mobility, accounting for its role as a molecular switch. We also show that natural mutations in human PPARgamma, associated with severe insulin resistance and diabetes mellitus, exhibit perturbations in the dynamic behavior of helix 12. Our findings provide the first direct observations of the mobility of helix 12 and suggest that the dynamic properties of this helix are key to the regulation of transcriptional activity. PMID- 12536207 TI - Germline transmission of RNAi in mice. PMID- 12536208 TI - Dissociated neural representations of intensity and valence in human olfaction. AB - Affective experience has been described in terms of two primary dimensions: intensity and valence. In the human brain, it is intrinsically difficult to dissociate the neural coding of these affective dimensions for visual and auditory stimuli, but such dissociation is more readily achieved in olfaction, where intensity and valence can be manipulated independently. Using event-related functional magnetic resonance imaging (fMRI), we found amygdala activation to be associated with intensity, and not valence, of odors. Activity in regions of orbitofrontal cortex, in contrast, were associated with valence independent of intensity. These findings show that distinct olfactory regions subserve the analysis of the degree and quality of olfactory stimulation, suggesting that the affective representations of intensity and valence draw upon dissociable neural substrates. PMID- 12536209 TI - Actin has a molecular scaffolding, not propulsive, role in presynaptic function. AB - We used actin tagged with enhanced green fluorescent protein (EGFP-actin) to characterize the distribution and dynamics of actin in living presynaptic terminals in rat CNS neurons. Actin was preferentially concentrated around--and appeared to surround--the presynaptic vesicle cluster. In resting terminals, approximately 30% of actin was found to be in a polymerized but dynamic state, with a remodeling time scale of approximately 20 s. During electrical activity, actin was further polymerized and recruited from nearby axonal regions to the regions surrounding vesicles. Treatment of terminals with the actin monomer sequestering agent latrunculin-A completely dispersed the actin network and abolished activity-dependent actin dynamics. We used a variety of methods to examine the role of actin in the presynaptic vesicle cycle. These data rule out a propulsive role for actin, either in maintaining the vesicle cluster or in guiding vesicle recycling. Instead, we propose that actin acts as a scaffolding system for regulatory molecules in the nerve terminal. PMID- 12536210 TI - Stop-signal inhibition disrupted by damage to right inferior frontal gyrus in humans. PMID- 12536211 TI - Capricious expression of cortical columns in the primate brain. PMID- 12536212 TI - Self-control during response conflict by human supplementary eye field. PMID- 12536213 TI - The eye is necessary for a circadian rhythm in the suprachiasmatic nucleus. PMID- 12536214 TI - PKA phosphorylation of AMPA receptor subunits controls synaptic trafficking underlying plasticity. AB - The regulated incorporation of AMPA receptors into synapses is important for synaptic plasticity. Here we examine the role of protein kinase A (PKA) in this process. We found that PKA phosphorylation of the AMPA receptor subunits GluR4 and GluR1 directly controlled the synaptic incorporation of AMPA receptors in organotypic slices from rat hippocampus. Activity-driven PKA phosphorylation of GluR4 was necessary and sufficient to relieve a retention interaction and drive receptors into synapses. In contrast, PKA phosphorylation of GluR1 and the activity of calcium/calmodulin-dependent kinase II (CaMKII) were both necessary for receptor incorporation. Thus, PKA phosphorylation of AMPA receptor subunits contributes to diverse mechanisms underlying synaptic plasticity. PMID- 12536215 TI - Integrating transcriptional and metabolite profiles to direct the engineering of lovastatin-producing fungal strains. AB - We describe a method to decipher the complex inter-relationships between metabolite production trends and gene expression events, and show how information gleaned from such studies can be applied to yield improved production strains. Genomic fragment microarrays were constructed for the Aspergillus terreus genome, and transcriptional profiles were generated from strains engineered to produce varying amounts of the medically significant natural product lovastatin. Metabolite detection methods were employed to quantify the polyketide-derived secondary metabolites lovastatin and (+)-geodin in broths from fermentations of the same strains. Association analysis of the resulting transcriptional and metabolic data sets provides mechanistic insight into the genetic and physiological control of lovastatin and (+)-geodin biosynthesis, and identifies novel components involved in the production of (+)-geodin, as well as other secondary metabolites. Furthermore, this analysis identifies specific tools, including promoters for reporter-based selection systems, that we employed to improve lovastatin production by A. terreus. PMID- 12536217 TI - Flow-cytometric isolation of human antibodies from a nonimmune Saccharomyces cerevisiae surface display library. AB - A nonimmune library of 10(9) human antibody scFv fragments has been cloned and expressed on the surface of yeast, and nanomolar-affinity scFvs routinely obtained by magnetic bead screening and flow-cytometric sorting. The yeast library can be amplified 10(10)-fold without measurable loss of clonal diversity, allowing its effectively indefinite expansion. The expression, stability, and antigen-binding properties of >50 isolated scFv clones were assessed directly on the yeast cell surface by immunofluorescent labeling and flow cytometry, obviating separate subcloning, expression, and purification steps and thereby expediting the isolation of novel affinity reagents. The ability to use multiplex library screening demonstrates the usefulness of this approach for high throughput antibody isolation for proteomics applications. PMID- 12536216 TI - A genomics-guided approach for discovering and expressing cryptic metabolic pathways. AB - Genome analysis of actinomycetes has revealed the presence of numerous cryptic gene clusters encoding putative natural products. These loci remain dormant until appropriate chemical or physical signals induce their expression. Here we demonstrate the use of a high-throughput genome scanning method to detect and analyze gene clusters involved in natural-product biosynthesis. This method was applied to uncover biosynthetic pathways encoding enediyne antitumor antibiotics in a variety of actinomycetes. Comparative analysis of five biosynthetic loci representative of the major structural classes of enediynes reveals the presence of a conserved cassette of five genes that includes a novel family of polyketide synthase (PKS). The enediyne PKS (PKSE) is proposed to be involved in the formation of the highly reactive chromophore ring structure (or "warhead") found in all enediynes. Genome scanning analysis indicates that the enediyne warhead cassette is widely dispersed among actinomycetes. We show that selective growth conditions can induce the expression of these loci, suggesting that the range of enediyne natural products may be much greater than previously thought. This technology can be used to increase the scope and diversity of natural-product discovery. PMID- 12536218 TI - Phosphorylated serine 199 of microtubule-associated protein tau is a neuronal epitope abundantly expressed in youth and an early marker of tau pathology. AB - Microtubule-associated protein tau is abnormally phosphorylated in many neurodegenerative disorders, and is the major component of neurofibrillary degeneration, a degenerating process with many biochemical phenotypes. The serine 199 (S199) residue of tau is phosphorylated at early and late stages of Alzheimer's disease (AD). We studied the immunohistochemical distribution of this phosphorylated epitope in AD and other neurodegenerative disorders, as well as in controls of different ages. The phosphorylated S199 (S199P) epitope was observed in tau lesions from numerous diseases with neurofibrillary degeneration. This epitope was found to be abundantly expressed in the hippocampus formation in childhood and in young adult brain samples, and more specifically in subsets of neurons vulnerable to neurodegeneration. Interestingly, our data suggests that S199P is particularly resistant to phosphatase activity occurring during post mortem delays. We suggest a peculiar and important role of the S199 residue as a qualitative indicator of the normal and pathological phosphorylation status of tau proteins. PMID- 12536219 TI - The relationship between extramotor ubiquitin-immunoreactive neuronal inclusions and dementia in motor neuron disease. AB - Dementia is thought to be an uncommon complication of motor neuron disease (MND). In addition to the characteristic motor system degeneration, pathological studies of MND patients with dementia have demonstrated changes in extramotor cortex; ubiquitin-immunoreactive inclusions are present in neocortical layer II neurons and hippocampal dentate granule cells. To examine how specifically this pathology is associated with dementia in MND, we performed ubiquitin immunohistochemistry on sections of hippocampus, prefrontal and temporal neocortex from 29 cases of MND, 10 with dementia and 19 with no clinical history of cognitive impairment. All cases with dementia had numerous ubiquitin-positive inclusions in dentate granule cells, whereas involvement of the neocortex was more variable. Six (32%) of the nondemented cases had ubiquitin pathology, which was similar to the demented cases in its morphology and distribution but of slightly less severe degree. These findings demonstrate that, although ubiquitinated inclusions in extramotor cortex are a consistent finding in MND with dementia, they are also common in MND in the absence of documented cognitive abnormalities. Such cases may either represent a subclinical stage of pathology or indicate that cognitive dysfunction is an underrecognized complication of MND. PMID- 12536220 TI - Influences of intranuclear inclusion on nuclear size - morphometric study on pontine neurons of neuronal intranuclear inclusion disease cases. AB - In looking for a possible influence of nuclear inclusions (NIs) on neurodegeneration in human brains, we quantified morphological features of pontine neurons of three unrelated cases of neuronal intranuclear inclusion disease (NIID) and five control cases. Cross-sectional area of each neuronal nucleus and the indices for its deformity (long axis/short axis and circularity index defined as deviation from the perfect circle) were measured on pontine sections and their relation to NIs was statistically analyzed. Cross-sectional area of neuronal nuclei harboring ubiquitin-immunopositive NIs was significantly larger (110.6+/-1.6 micro m(2), mean +/- SE), while that of nuclei not harboring NIs was smaller (77.8+/-1.5 micro m(2)) than that in controls (90.5+/-0.7 micro m(2)). This difference remained significant even when the cross-sectional area occupied by NIs was subtracted from that of the nucleus harboring the NI (97.4+/ 1.5 micro m(2)). This could hardly be explained if nuclear shrinkage is accelerated in the presence of NI. On the contrary, NI formation in pontine neurons of NIID might be linked, either directly or indirectly, to a mechanism, which counteracts rather than accelerates nuclear shrinkage. Because nuclear deformity was apparent even in neurons with NIs, whose nuclei were significantly larger than controls, the nuclear deformity is not secondary to its shrinkage and represents another aspect of neurodegeneration independent of nuclear shrinkage. Association of NIs to neurons of larger nuclear size in NIID brain indicates that NIs are not necessarily toxic to neurons. PMID- 12536221 TI - Anti-neurofilament antibodies in neuropathy with monoclonal gammopathy of undetermined significance produce experimental motor nerve conduction block. AB - Elevated levels of serum antibodies to neurofilament proteins have been associated with a variety of neurological diseases, including autoimmune disorders such as neuropathy with monoclonal gammopathy of undetermined significance (MGUS). The pathological significance of anti-neurofilament antibodies in sera of affected patients, however, remains unclear. In this study, we report our findings of polyclonal antibodies in sera from 4 of 16 IgG MGUS neuropathy patients that react strongly on immunoblot with a high molecular weight neurofilament protein (NFH). The effect of anti-NFH polyclonal antibody on peripheral nerve function was tested in vivo by intraneural injection. Sera containing anti-NFH antibody, but not sera from age-matched control subjects, injected into the endoneurium of rat sciatic nerve significantly attenuated proximal-evoked motor nerve compound muscle action potential (CMAP) amplitudes in a complement-dependent manner. In contrast, ankle-evoked CMAP amplitudes were unaffected by intraneural injection of sera containing anti-NFH antibody. Anti NFH serum-injected nerves showed changes in both axon caliber (shrinkage) and myelin ultrastructure (vesiculation and ovoid formation), suggestive of intramyelinic edema. Preincubation of sera containing anti-NFH antibody with purified NFH protein abolished immunoreactivity to NFH protein and neutralized the serum-mediated toxicity. The data suggest that anti-NFH polyclonal antibodies occurring in sera of some patients with IgG MGUS neuropathy may elicit peripheral nerve conduction block independent of the patients' IgG paraprotein. Anti-neural polyclonal antibodies in sera of IgG MGUS neuropathy patients may have a greater pathological significance than previously anticipated. PMID- 12536222 TI - Cholinergic neuronal loss in the basal forebrain and mesopontine tegmentum of progressive supranuclear palsy and corticobasal degeneration. AB - Progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD) are clinically characterized by atypical parkinsonism and cognitive disorders and classified in the same histopathological category showing neuronal and glial neurofibrillary tangles (NFTs). To characterize the vulnerability of the forebrain and midbrain cholinergic systems in PSP and CBD, we performed a comparative study of cholinergic neuronal changes in the nucleus basalis of Meynert (NBM) and the laterodorsal tegmental and pedunculopontine tegmental nuclei (LdtgN and PptgN) of brains obtained at autopsy (three cases of PSP, one case of CBD and six cases without neurological diseases) by immunohistochemistry for choline acetyltransferase (ChAT) and Gallyas-Braak staining. In the NBM, the number of neurons and the ChAT-positivity rate of remaining neurons were decreased more in CBD than PSP. On the other hand, in the PptgN and LdtgN neurons were reduced much more, and more NFTs were observed in PSP than CBD. PSP showed a severe decrease of neurons and the ChAT-immunopositive neurons in the LdtgN but less in the PptgN. In CBD, there was a mild deletion of the ChAT-immunostained neurons in the PptgN, but not in the LdtgN. In PSP, cholinergic neurons in the LdtgN are likely to be more vulnerable than PptgN and NBM. PMID- 12536223 TI - Breakdown of the blood-brain barrier and neuropathological changes induced by Phoneutria nigriventer spider venom. AB - The blood-brain barrier (BBB) is responsible for selective flux of substances between blood and brain. The selective permeability of the BBB is crucial for the maintenance of the brain microenvironment homeostasis, and alterations in the barrier may be involved in many pathophysiological processes. Phoneutria nigriventer armed spider venom produces excitatory signals and symptoms in humans, and its recognized neurotoxic action suggests a potential ability to alter BBB permeability. The aim of the present study was to investigate the capacity of P. nigriventer venom (PNV) in promoting BBB breakdown in adult rats. After intravenous injection of 850 micro g/kg of the whole venom, BBB lesions were evaluated after 18 h to 9 days by ultrastructural methods using the extracellular tracer lanthanum nitrate. Clinical signs and symptoms of rats showed acute neurotoxicity, with some of the animals presenting convulsions, but which were clinically resolved by 12 h post-envenoming. The results showed that PNV is able to increase BBB permeability, particularly in the hippocampus. Changes were first detected in arterioles and post-capillary venules 18 h to 5 days after venom inoculation. The increased permeation of the extracellular tracer peaked on day 1, representing about 42% of the examined vessels (P<0.01). This appeared to occur by both transendothelial and intercellular routes, i.e., by pinocytic transport and through interendothelial junctions. Concomitantly, the surrounding tissue showed vasogenic edema and swollen astrocytic processes, without inflammatory infiltrates. The peak of the edema occurrence was observed on day 3, in about 60% of the vessels (P<0.001). Enhanced capillary permeability was observed on day 9, and affected 36% of all capillaries (P<0.05). The affected capillaries were characterized by increased number of pinocytotic vesicles, which, in addition, were filled with the extracellular tracer, but without visible transport through the interendothelial pathway. This study demonstrates that systemic PNV inoculation induces BBB breakdown through trans- and paracellular routes. It is concluded that BBB breakdown is an event not associated with the acute neurotoxicity exhibited by the rats. PMID- 12536224 TI - Amyloid beta plaque-associated proteins C1q and SAP enhance the Abeta1-42 peptide induced cytokine secretion by adult human microglia in vitro. AB - Pro-inflammatory cytokines released by activated microglia could be a driving force in Alzheimer's disease (AD) pathology. We evaluated whether the presence of complement factor C1q and serum amyloid P component (SAP) in Abeta deposits is related to microglial activation. Activated microglia accumulate in SAP- and C1q immunoreactive fibrillar amyloid beta (Abeta) plaques in AD temporal cortex. No clustered microglia are seen in SAP- and C1q-positive circumscript, non fibrillar, tau-negative Abeta plaques in AD caudate nucleus and non-demented control temporal cortex. In addition, no clustered microglia were observed in C1q and SAP-negative, irregular shaped, diffuse plaques in AD caudate nucleus and in non-demented control temporal cortex, which suggests that microglia are attracted and activated in Abeta deposits of certain fibrillarity that, in addition, have fixed SAP and C1q. Therefore, the effects of Abeta(1-42), SAP and C1q on cytokine secretion by human postmortem microglia in vitro were assessed. Abeta(1-42) alone had little to no effect. Abeta(1-42) peptides in combination with C1q or C1q and SAP increased microglial interleukin (IL)-6 secretion four- and eightfold, respectively. Tumor necrosis factor (TNF)-alpha, as well as intracellular IL 1alpha and IL-1beta levels, also increased upon exposure of microglia to Abeta(1 42)-SAP-C1q complexes. Combined with earlier findings, that amyloid and activated microglia accumulate at a relatively early stage of cognitive decline in AD patients, this suggests that clustering of activated, cytokine-secreting microglia in SAP- and C1q-containing Abeta deposits precedes neurodegenerative changes in AD, and thus may provide a "therapeutic window". PMID- 12536225 TI - Distribution, progression and chemical composition of cortical amyloid-beta deposits in aged rhesus monkeys: similarities to the human. AB - A comprehensive investigation of the incidence, distribution, progression and chemical composition of Abeta deposits in the brains of two young (5 years) and seven aged (25-30 years) rhesus monkeys was conducted to determine the similarity of this phenomenon to that in the human. The brains of the young rhesus were devoid of Abeta deposits. In contrast, Abeta deposits were observed within the cerebral cortex of all aged animals. In animals with mild Abeta burden, deposits were observed primarily in association cortical zones. In animals with moderate Abeta burden, many paralimbic cortical zones also contained Abeta deposits. Finally, in an animal with a heavy burden of Abeta, core limbic cortical zones were also involved. The primary sensory and motor cortices were relatively free of Abeta deposits. A higher proportion of plaques contained Abeta40 as compared with Abeta42. Abeta deposits contained a number of other constituents. Cholinesterases were present in nearly 50% of plaques and displayed the exact same biochemical characteristics as those in the human. Nearly 20% of Abeta deposits also contained apolipoprotein E and a smaller proportion contained heparin sulfate proteoglycans and alpha1-anti-chymotrypsin. The latter three chemicals were present in many compact plaques. These results indicate that the distribution, progression and chemical composition of plaques in the aged rhesus monkey display many similarities to those observed in the aged human and Alzheimer's disease. Therefore, despite some differences from the human, the aged rhesus may be a good model for studies of the pathological effects of Abeta in the primate brain. PMID- 12536226 TI - Human GLUT5 immunolabeling is useful for evaluating microglial status in neuropathological study using paraffin sections. AB - In neuropathological studies it is important to detect both resting and reactive microglia in paraffin sections. We examined the usefulness of human (h) GLUT5, a glucose transporter, as a microglial marker. We produced an hGLUT5 antibody against its C-terminal sequence and stained human brain tissue sections. The hGLUT5 antibody consistently stained microglia in cryostat sections. In paraffin sections fixed with formalin, paraformaldehyde or ethanol, both resting and reactive microglia were stained; the latter were stained more intensely than the former. The hGLUT5 and glial fibrillary acidic protein labeling did not overlap each other in double immunofluorescence analyses. Oligodendrocytes, perivascular cells, choroid plexus epithelium and ependymal cell were negative for hGLUT5. Even after 1-month fixation in formalin, the hGLUT5 antibody stained microglia well. Microwave pretreatment enhanced the immunoreactivity of hGLUT5. As compared with other microglial markers, KP-1, KiM1p, CR3.43 and RCA-1, the hGLUT5 antibody could be considered good morphological marker. hGLUT5 immunolabeling clearly showed the detailed microglial processes, whereas immunolabeling with Ki-M1P and KP-1 showed cytoplasmic granules, and it was difficult to trace the microglial processes. The hGLUT5 antibody stained both resting and reactive microglia, whereas CR3.43 stained only reactive microglia, and RCA-1 labeled microvessels more intensely than microglia. Thus, hGLUT5 is a marker that is suitable for routine histopathological staining procedures. PMID- 12536228 TI - Characterization of eosinophilic hyaline droplets in schwannoma. AB - In schwannoma, degenerative structures, such as bizarre hyperchromatic nuclei, lipofuscin deposition, hemorrhage, edema, etc., are frequently encountered and are regarded as hallmarks of the benign nature of the neoplasm. Hyaline globules or eosinophilic hyaline droplets (EHDs) have been observed in a large variety of tumors, but they are rare histological manifestations of schwannoma, and the details have not been described as yet. We reviewed 171 cases of schwannoma of conventional histology. We classified them based on the sites where they occurred as follows: 43 acoustic nerves, 9 other cranial nerves, 30 spinal nerve roots, and 89 soft parts. We found EHDs in 8/43, 1/9, 1/30, 2/89 cases, respectively. The droplets were located in both the perikarya and processes. Ultrastructurally, they were electron-dense, round to ovoid organelles, and indistinguishable from secondary lysosomes. MIB-1 immunolabeling of the droplet-bearing cells was almost negative; however, they maintained a viable nuclear appearance without karyorrhexis, pyknosis or apoptotic changes. We therefore conclude that EHD is a new member of degenerative structures in schwannoma, and a hallmark of low growth potential. Furthermore, their higher frequency in acoustic schwannoma may suggest their distinct nature from those originated in the other sites. Possible relevance of EHDs and cellular senescence is also discussed. PMID- 12536227 TI - Glial involvement in diffuse Lewy body disease. AB - Diffuse Lewy body disease (DLBD) is characterized by the presence of Lewy bodies (LB) in the neurons and neurites of cortical, subcortical, and brain stem structures. Recently, alpha-synuclein (alphaS) has been found to be a central constituent of LB. In DLBD, abnormal accumulation of alphaS has been reported in both neurons and glia, but studies on glial lesions in DLBD have been limited. We examined in detail the constituents and distribution of glial lesions in eight patients with DLBD and report the pathogenesis of the glial lesions. alphaS positive neuronal cytoplasmic inclusions (NI), neuropil threads (NT), and coiled bodies (CB) showed similar immunostaining profiles. Without pretreatment, NI, NT, and CB were detected by all antibodies against alphaS. The immunostaining profile of star-like astrocytes (SLA) was quite different from those of NI, NT, and CB. A few SLA were stained by an antibody against the non-Abeta component portion of alphaS without pretreatment, but formic acid pretreatment dramatically enhanced SLA immunoreactivity. SLA and CB were found in all eight brains with DLBD. SLA were scarce in the brain stem, but there were hundreds of SLA per visual field at x100 magnification in the temporal cortex of most cases, while CB were found diffusely in both the cerebral cortex and brain stem, similar to NI. This suggests that the pathogenesis of SLA is different from those of NI and CB. PMID- 12536229 TI - Synphilin in normal human brains and in synucleinopathies: studies with new antibodies. AB - Mutations in the gene encoding alpha-synuclein (alpha-syn) have recently been linked to rare hereditary forms of Parkinson's disease. A yeast two-hybrid screen with alpha-synuclein (alpha-syn) identified synphilin as an alpha-syn-interacting protein, potentially implicating synphilin in the pathogenesis of synucleinopathies. Co-transfection of synphilin and the central (NAC) region of alpha-syn in HEH293 cells resulted in synuclein inclusions. Furthermore, synphilin immunoreactivity has been observed in Lewy bodies (LBs) and glial cytoplasmic inclusions of synucleinopathies. To further characterize synphilin, we utilized two new anti-synphilin antibodies for biochemical and immunohistochemical studies in normal and disease brain tissues. In normal brain tissue, synphilin localized predominantly to large neurons, such as substantia nigra neurons, hippocampal pyramidal and cerebellar Purkinje cells. However, in a few pathological cases synphilin immunoreactivity was present in glial cells and a small percentage of cortical and nigral LBs. In brain extracts, synphilin was observed primarily as a 90-kDa band but protein bands of 50 and 65 kDa were also present in both soluble (high salt) and lipid (Triton X-100) fractions. Additionally, less abundant higher molecular mass species, including a 120-kDa band of similar size to that of synphilin expressed in transiently transfected cells were recovered in 8 M urea-solubilized pellets after sequential extraction of brain tissue with buffers of increasing strengths. The presence of the synphilin of higher molecular mass was detected regardless of alpha-syn pathology and may represent an immature form of synphilin. Thus, although synphilin may be an alpha-syn-interacting protein present in some alpha-syn lesions, it still remains to be determined whether synphilin plays a critical role in mechanisms of brain degeneration in human synucleinopathies. PMID- 12536230 TI - Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) with hypertrophic spinal radiculopathy mimicking neurofibromatosis. AB - This report illustrates a case of chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) masquerading as neurofibromatosis due to multifocal enlargements of spinal nerve roots. The patient initially complained of intermittent numbness of the hands and leg weakness at age 62. Nerve conduction velocities were reported to be abnormally slow, suggesting a diagnosis of demyelinating neuropathy. A complaint of progressive lower back pain 4 years later prompted a lumbar CT myelogram, which demonstrated bilateral nerve root enlargements. A biopsy of an enlarged lumbar root obtained during decompressive laminectomy was interpreted as consistent with a plexiform neurofibroma. He suffered recurrent paraparesis, at times with a sensory level indicating spinal cord compression, which responded to corticosteroid therapy. An autopsy 15 years after the onset of symptoms revealed hypertrophic radiculopathy and peripheral neuropathy due to CIDP with no evidence of neurofibromatosis. This case illustrates how the hypertrophic neuropathy accompanying CIDP can be mistaken for neurofibromatosis. PMID- 12536232 TI - Chiari anomaly type II without cerebellar herniation. AB - This is the first neuropathological documentation of a less known variant of Chiari anomaly type II without cerebellar herniation. It should be established that Chiari anomalies type II and type I belong to the same category of the basicranio-cervical hypoplasia and are not a primary CNS malformation. PMID- 12536231 TI - Lewy bodies in Betz cells of the motor cortex in a patient with Parkinson's disease. AB - Lewy bodies (LBs) were observed in the giant pyramidal cells of Betz in the motor cortex of a patient with Parkinson's disease (PD) of 8-year duration. The patient had shown typical clinical features of PD. No dementia or pyramidal tract signs had been observed. The LBs, found in 8 of 747 Betz cells counted (1.1%), appeared as homogeneous or laminated, spherical inclusions with a clear surrounding halo and were strongly immunoreactive for alpha-synuclein. To our knowledge, this is the first demonstration of LBs in Betz cells. Considering the significant loss of neurons in the predilection sites for LBs, it is possible that in this patient, the motor cortex was also involved in the disease process of PD. PMID- 12536233 TI - Response to B.s. Kasper, acta neuropathol (2002) 103:307. PMID- 12536234 TI - Analysis of HLA expression in human tumor tissues. AB - Cancer cells can be detected and destroyed by cytotoxic T lymphocytes in many experimental tumor systems, and--as has been well-documented--in some human tumors. In humans however, most diagnosed tumors are not eliminated by T cells but grow steadily, invading and metastasizing until the host is destroyed. Evidence is accumulating that progressive tumor growth occurs not because the immune system is defective or deteriorated, but because the cancer cell is capable of developing a variety of strategies to escape immune recognition. In addition, cancer cells acquire new biological properties to generate invasive capacity in order to migrate and colonize new tissues. Major histocompatibility complex (MHC) antigens are molecules that are specialized in communicating with the T cell receptor and natural killer (NK) cell ligands. With the former, they use the interaction with peptides derived from processed cellular and exogenous proteins to monitor self and non-self status. With the latter, they determine the degree of activation and killing capacity of NK cells by interacting with NK receptors. Any change in the MHC profile of tumor cells (including classical and nonclassical MHC molecules) may therefore have a profound influence on the immune recognition and immune rejection of cancer cells. We have reviewed the data from our laboratory and other groups, and have presented a standardized procedure for analyzing the MHC profile of human tumors with special emphasis on the quality and laboratory use of the material obtained from microdissected tumor samples. Appropriate tissue processing is of particular relevance, since it is not possible to obtain tumor cell lines from most patients. Oncologists require rapid information on the MHC profile of the tumor if gene therapy is envisaged to restore normal MHC class I gene expression. PMID- 12536235 TI - Intra-arterial administration of TNF-alpha followed by arterial ablation is an effective therapy for a regionally confined TNF-resistant rat mammary adenocarcinoma. AB - Tumor necrosis factor-alpha (TNF-alpha) is an immunomodulatory cytokine that has exhibited anti-tumor activity in a variety of experimental systems. However, the toxicities associated with systemic administration of TNF-alpha have limited its clinical utility and have led to the investigation of targeted delivery techniques with the ability to present the TNF-alpha dose directly to the vascular bed of the tumor. The intra-arterial (IA) administration of TNF-alpha to patients with liver metastases represents one such approach, and recent work suggests that subsequent ablation of the tumor's arterial supply via embolization may enhance the efficacy of intra-arterial treatments (hepatic chemoembolization). The present study was undertaken to test the hypothesis that IA administration of TNF-alpha is superior to the intravenous (IV) route for inhibition of tumor growth in a regionally confined rat mammary adenocarcinoma model that provides for ablation of the arterial supply to the tumor following cytokine therapy. Rats bearing hind limb mammary adenocarcinomas received single IA or IV infusions of 8 x 10(5), 1 x 10(6), and 1.5 x 10(6) units of TNF-alpha via the common femoral artery (CFA) followed 1 h later by ligation of the artery. Control animals received either no treatment or IA infusion of 2% normal rat serum (NRS) followed by ipsilateral CFA ligation. Tumor size was measured every other day after treatment. Tumor growth inhibition occurred in the first 5 to 10 days after treatment. IV administration of TNF-alpha did not result in visual tumor necrosis or significant reduction in the rate of tumor growth. IA administration of TNF-alpha resulted in statistically significant diminution of tumor size as compared to untreated controls and animals receiving IA 2% normal rat serum (NRS; P<0.05 at days 6, 8 and 10), regardless of the dose employed. The maximum growth inhibition with IA TNF-alpha was a 91% reduction in tumor volume that was achieved with a dose of 1 x 10(6) U TNF-alpha. These results demonstrate improved anti-tumor activity with the IA administration of TNF-alpha over the IV route in a regionally confined mammary adenocarcinoma. IA administration of biologic response modifiers like TNF-alpha may therefore be a useful approach for the hepatic chemoembolization of breast adenocarcinomas metastatic to the liver. PMID- 12536236 TI - Rapid generation of antigen-presenting cells from leukaemic blasts in acute myeloid leukaemia. AB - The ability of acute myeloid leukaemia (AML) cells to acquire dendritic cell (DC) like characteristics in vitro with a rapid culture method based either on the phorbol ester PMA or calcium ionophores has been studied in comparison to conventional AML-DC cultures with the cytokines granulocyte-macrophage colony stimulating factor (GM-CSF), tumour necrosis factor-alpha (TNF-alpha), interleukin-3 (IL-3), SCF, FLT3-L and IL-4. In all AML patients, antigen presenting cells (APC) could be generated from leukaemic cells in 2 days by incubation with PMA or calcium ionophore (A23187 or ionomycin) in the presence as well as in the absence of IL-4. In 30 out of 36 patients APC could be generated after 2 weeks of culture in cytokine-enriched medium. AML-APC cultured with PMA or calcium ionophores immunophenotypically and functionally were at a more mature stage than those cultured in cytokine-enriched medium. The most mature APC were generated by calcium ionophore A23187 plus IL-4, as evidenced by the higher expression of CD40, CD80, CD86 and HLA-DR. Autologous T cell mediated cytotoxicity towards AML blast cells in vitro was observed in 2 cases tested. The persistence of cytogenetic abnormalities confirmed the leukaemic origin of the AML-APC. The generation of AML-APC was possible from freshly isolated as well as cryopreserved material. Our data show that generation of sufficient AML-APC by A23187 plus IL-4 is feasible, for vaccination purposes, in approximately 70% of AML specimens, offering a time-saving and cost-effective approach in preparing anti-leukaemia vaccines. PMID- 12536237 TI - Interferon-gamma secretion and perforin expression are impaired in CD8+ T lymphocytes from patients with undifferentiated carcinoma of nasopharyngeal type. AB - An efficent antitumor and antiviral cellular immune response requires optimal interferon-gamma (IFN-gamma) secretion and perforin expression in CD8(+) T cells. The aim of this study was to define whether CD4(+) and CD8(+) T cells from patients with undifferentiated carcinoma of nasopharyngeal type (UCNT), a tumor regularly associated with the Epstein-Barr virus (EBV), have abnormal phenotype profiles, cytokine production, perforin and CD3-zeta expressions. Our data showed that CD4 and CD8 subset distribution was not grossly altered in the peripheral blood of UCNT patients, while tumor biopsies contained an increased proportion of CD8(+) T cells. The analysis of the CD4(+) subset showed a defect in interleukin 2 (IL-2) production and a moderate increase of IL-10 production, a situation consistent with a Th1/Th2 imbalance. We have also demonstrated that CD8(+) lymphocytes from UCNT patients had a marked impairment of IFN-gamma secretion and perforin expression. This impairment was not related to the presence of detectable EBV DNA in the plasma. In UCNT patients, the blockade of the perforin pathway and of IFN-gamma production may constitute important mechanisms for immune escape by the tumor and for impaired control of EBV replication. PMID- 12536238 TI - A series of immune responses leading to the induction of T cell IL-12/IL-18 responsiveness in patients with relatively large tumor burdens. AB - The induction of interleukin-12 (IL-12) responsiveness in T cells depends on T cell receptor (TCR) triggering, and is regarded as a parameter of recently TCR sensitized T cells. Here, we investigated whether IL-12 responsiveness could be detected in freshly prepared T cells from tumor-bearing patients, and if so whether such patients exhibited additional immunological parameters related to IL 12 responsiveness. CD4(+) and CD8(+) T cell populations from an appreciable proportion of tumor-bearing patients exhibited high levels of IL-12 responsiveness as evaluated by IL-12-stimulated interferon-gamma (IFN-gamma) production. T cell populations with high IL-12 responsiveness were observed in the group of patients with moderate to large tumor mass or tumor metastases rather than in patients with small tumors. The frequency of such a T cell population was also lower in post-surgery tumor-free patients, showing the correlation between IL-12 responsiveness and the presence of a certain extent of tumor burden. More importantly, a higher incidence of IL-12 responsiveness was observed in tumor-bearing patients exhibiting detectable plasma IL-12 levels, and correlated with IL-18 responsiveness. T cell IL-12 and IL-18 responsiveness is induced by TCR triggering and subsequent IL-12 stimulation respectively. Furthermore, TCR-triggered T cells stimulate antigen-presenting cells (APC) to produce IL-12. Therefore, the present observations suggest that an immune response loop from TCR sensitization to the induction of IL-12/IL-18 responsiveness via IL-12 production operates in tumor-bearing patients, particularly in those with relatively large tumor burdens. PMID- 12536239 TI - Clinical response after intradermal immature dendritic cell vaccination in metastatic melanoma is associated with immune response to particulate antigen. AB - Metastatic melanoma is poorly responsive to treatment, and immunotherapeutic approaches are potentially beneficial. Predictors of clinical response are needed to identify suitable patients. We sought factors associated with melanoma specific clinical response following intradermal vaccination with autologous melanoma peptide and particulate hepatitis B antigen (HBsAg)-exposed immature monocyte-derived dendritic cells (MDDC). Nineteen patients with metastatic melanoma received a maximum of 8, 2-weekly vaccinations of DC, exposed to HBsAg in addition to autologous melanoma peptides. A further 3 patients received an otherwise identical vaccine that did not include HBsAg. Patients were assessed 1 2 monthly for safety, disease volume, and cellular responses to HBsAg and melanoma peptide. There was no significant toxicity. Of 19 patients receiving HBsAg-exposed DC, 9 primed or boosted a cellular response to HBsAg, and 10 showed no HBsAg response. HBsAg-specific responses were associated with in vitro T cell responses to melanoma peptides and to phytohemagglutinin (PHA). Zero out of 10 non-HBsAg-responding and 4/9 HBsAg-responding patients achieved objective melanoma-specific clinical responses or disease stabilization - 1 complete and 2 partial responses and 1 case of stable disease ( P=0.018). Development of melanoma-specific cellular immunity and T cell responsiveness to mitogen were greater in the group of patients responding to HBsAg. Therefore stimulation of an immune response to nominal particulate antigen was necessary when presented by melanoma peptide-exposed immature DC, to achieve clinical responses in metastatic melanoma. Since general immune competence may be a determinant of treatment response, it should be assessed in future trials on DC immunotherapy. PMID- 12536240 TI - Selective reduction of natural killer cells and T cells expressing inhibitory receptors for MHC class I in the livers of patients with hepatic malignancy. AB - Natural killer (NK) and CD56(+) T cells are thought to play a central role in antitumour immunity. Their cytolytic activities are controlled by a variety of receptors including CD94 and killer immunoglobulin-like receptors (KIR), which bind to major histocompatibility complex (MHC) class I molecules on target cells and mediate cell activation or inhibition. We have examined the numbers, phenotypes and antitumour cytotoxic functions of hepatic NK and CD56(+) T cells isolated from 22 patients with hepatic malignancy and 19 healthy donors. Flow cytometry revealed that NK cell numbers were increased among hepatic mononuclear cells in malignancy compared to histologically normal livers (mean: 38% vs 27%; P=0.03), but CD56(+) T cell numbers were not (28% vs 27%). NK cells and CD56(+) T cells from tumour-bearing livers exhibited lymphokine-activated killing of K562 targets and T cell receptor-mediated lysis of P815 cells. The expression of CD94 and the KIR isotypes CD158a, CD158b and KIR3DL1 by CD56(+) T cells and NK cells was significantly and consistently reduced in tumour-bearing livers compared to healthy livers ( P<0.05 in all cases). Simultaneous ligation of CD158a, CD158b and KIR3DL1 caused an overall partial inhibition of CD56(+) T cell cytotoxic activity, suggesting that the observed reductions in KIR(+) cell numbers in malignancy are likely to lead to enhanced cytotoxicity. Our results suggest that, while hepatic CD56(+) T cells are not expanded in malignancy, downregulation of KIR and CD94 expression may be a mechanism by which the hepatic immune system can be activated to facilitate tumour rejection. PMID- 12536242 TI - Respiratory motion artifacts on PET emission images obtained using CT attenuation correction on PET-CT. AB - PET-CT scanners allow generation of transmission maps from CT. The use of CT attenuation correction (CTAC) instead of germanium-68 attenuation correction (Ge AC) might be expected to cause artifacts on reconstructed emission images if differences in respiratory status exist between the two methods of attenuation correction. The aim of this study was to evaluate for possible respiratory motion artifacts (RMA) in PET images attenuation corrected with CT from PET-CT in clinical patients. PET-CT scans were performed using a Discovery LS PET-CT system in 50 consecutive patients (23 males, 27 females; mean age 58.2 years) with known or suspected malignancy. Both CTAC and Ge AC transmission data obtained during free tidal breathing were used to correct PET emission images. Cold artifacts at the interface of the lungs and diaphragm, believed to be due to respiratory motion (RMA), that were seen on CTAC images but not on the Ge AC images were evaluated qualitatively on a four-point scale (0, no artifact; 1, mild artifact; 2, moderate artifact; 3, severe artifact). RMA was also measured for height. Curvilinear cold artifacts paralleling the dome of the diaphragm at the lung/diaphragm interface were noted on 84% of PET-CT image acquisitions and were not seen on the (68)Ge-corrected images; however, these artifacts were infrequently severe. In conclusion, RMA of varying magnitude were noted in most of our patients as a curvilinear cold area at the lung/diaphragm interface, but were not diagnostically problematic in these patients. PMID- 12536241 TI - Different regulation of PKC isoenzymes and MAPK by PSK and IL-2 in the proliferative and cytotoxic activities of the NKL human natural killer cell line. AB - The activation of natural killer (NK) cells and induction of cytotoxicity are complex processes whose molecular mechanisms have not been clearly elucidated. Stimulation of the NKL human NK cell line with interleukin-2 (IL-2) or protein bound polysaccharide K (PSK) leads to sustained growth and cytolytic activity in comparison to unstimulated NKL cells. Our previous results shown that IL-2 and PSK regulate different nuclear transcription factors in NKL cells, and that the signal transduction pathway used by these inducers is different. To determine the molecular basis for the different action of IL-2 and PSK, we investigated the upstream effects generated in human NKL cells by IL-2 and PSK on protein kinase C (PKC) isoenzymes and mitogen-activated protein kinases (MAPK). Here we report the profile of unstimulated NKL cells as: PKCbeta>PKCalpha>PKCdelta =PKCepsilon. The PKCeta form was not expressed. The effects of PSK and IL-2 on these isoenzymes were different. IL-2 increased the expression of PKCalpha, PKCdelta and PKCepsilon, whereas PSK decreased the expression of PKCalpha, and also increased PKCdelta and PKCepsilon to higher levels than did IL-2. In MAPK expression we found that unstimulated NKL cells have the following profile: ERK2>ERK6>p38gamma>p38beta>ERK1. ERK3, ERK3 rel, ERK5/ERK4 and p38delta were not expressed. IL-2 decreased the expression of ERK2, whereas PSK did not, and both agents increased the expression of ERK3. These results shown that PSK and IL-2 produce different variations in PKC isoenzymes and MAPK in NKL cells. PMID- 12536243 TI - In vitro and in vivo study of 99mTc-MIBI encapsulated in PEG-liposomes: a promising radiotracer for tumour imaging. AB - Encapsulation of technetium-99m sestamibi ((99m)Tc-MIBI) in polyethyleneglycol liposomes ((99m)Tc-MIBI-PEG-liposomes) could extend the duration of its circulation in blood and alter its biodistribution, enabling its concentration in tumours to be increased. An original method to encapsulate (99m)Tc-MIBI in PEG liposomes is described. The (99m)Tc-MIBI-PEG-liposomes were compared with free (99m)Tc-MIBI with respect to (a) tumour availability (b) ability to distinguish between chemotherapy-sensitive and -resistant cells and (c) uptake ratio in tumour imaging. PEG-liposomal systems composed of distearoylphosphatidylcholine/cholesterol/PEG(2000)-distearoyl phosphatidylethanolamine and lissamine-rhodamine B-labelled liposomes were used. The encapsulation of (99m)Tc-MIBI in liposomes was achieved using the K(+) diffusion potential method. We compared the uptake of free versus encapsulated (99m)Tc-MIBI by sensitive and resistant erythroleukaemia (K562) and breast tumour (MCF-7ras) cells. To assess the internalisation of these liposomes into cells, rhodamine B-labelled PEG-liposomes were used and visualised by fluorescence microscopy. Biodistribution and imaging characteristics of encapsulated and free radiotracer were determined in rats and tumour-bearing nude mice. The efficiency of (99m)Tc-MIBI encapsulation in PEG-liposomes was 50+/-5%. Use of (99m)Tc-MIBI PEG-liposomes did not impair the ability of this tracer to distinguish between chemotherapy-sensitive and -resistant tumour cells; the percentage of radioactivity accumulated in the sensitive K562 cells was 1.24+/-0.04%, as compared with 0.41+/-0.04% in the resistant K562 cells. One hour post injection in rats, PEG-liposomes showed a ten times higher activity in blood than free (99m)Tc-MIBI, whereas activity of free (99m)Tc-MIBI in kidneys and bladder was markedly higher than that of encapsulated (99m)Tc-MIBI, indicating faster clearance of the free radiotracer. In the (MCF7-ras)-bearing nude mice, PEG liposome uptake in tumour was two times that of free (99m)Tc-MIBI. Summarising, the (99m)Tc-MIBI-PEG-liposomes demonstrated a longer blood circulation time, enabled distinction between chemotherapy-sensitive and -resistant cells and improved tumour to background contrast in in vivo imaging. (99m)Tc-MIBI-PEG liposomes therefore show promising potential for tumour imaging. PMID- 12536244 TI - Performance evaluation of the microPET R4 PET scanner for rodents. AB - The microPET R4 scanner is a dedicated positron emission tomograph (PET) for studies of rodents. A number of scanner parameters such as spatial resolution, sensitivity, scatter, and count rate performance were determined in this work, which showed that the microPET R4 is a suitable PET scanner for small animals like mice and rats. In the center of the field of view (FOV) a maximal sensitivity of 43.66 cps/kBq for a centered point source was calculated from a measurement with a germanium-68 line source within an energy widow of 250-750 keV. A spatial resolution of 1.85 mm full-width at half-maximum (FWHM) in the axial direction and 1.66 mm FWHM in the transaxial direction was measured in the center with a 1-mm-diameter sodium-22 point source. Within the inner 20 mm of the FOV the volumetric resolution is better than 15.6 micro l, corresponding to a linear resolution of less than 2.5 mm in all three dimensions. Images of a high resolution phantom and from mice and rat studies illustrate the good performance of the scanner. A maximal noise equivalent count rate (NECR) was reached at 174 kcps for a mouse phantom and at 93 kcps for a rat phantom (energy window 250-750 keV). Scatter fractions were measured between 0.30 and 0.42 for an energy window of 250-750 keV and phantom diameters similar to mice and rats. A comparison with the microPET P4 model for primates illustrates the gain in sensitivity due to a smaller detector ring diameter but also the changes in NECR. PMID- 12536245 TI - Radionuclide voiding patterns in children with vesicoureteral reflux. AB - The purpose of this study was to evaluate bladder function in children with vesicoureteral reflux (VUR) by means of indirect radionuclide cystography (IRNC), and to investigate whether IRNC can identify those children with voiding dysfunction. The study enrolled 74 neurologically intact children, 14 boys and 60 girls aged 2-14 years, with VUR documented using contrast micturating cystourethrography as the initial method. In all patients, IRNC was performed using technetium-99m diethylene triamine penta-acetate (DTPA). Based on the urodynamic findings, three groups were distinguished: a group with VUR and normal urodynamic findings ( n=27), a group with VUR and detrusor hyper-reflexia ( n=43) and a group with VUR and detrusor-sphincter dyssynergia ( n=4). A control group comprised 64 healthy children, aged 2-13 years, without any symptoms of lower urinary tract dysfunction. The dynamics of bladder emptying were studied in the posterior view after intravenous injection of 37 MBq/10 kg b.w. DTPA, with acquisition of 90 2-s frames during voiding. The parameters evaluated were: voided urine volume (VV), bladder capacity, functional bladder capacity (FBC), residual urine, voiding time, average flow rate, peak flow rate (PFR) and ejection fraction (EF). With regard to the final urodynamic diagnosis, FBC, PFR and EF were found to be significant IRNC predictor variables using the logistic regression method. If abnormality on at least two of the three significant predictor variables was taken as the criterion of voiding dysfunction, the overall sensitivity, specificity and accuracy of IRNC in the detection of voiding dysfunction were 81%, 78% and 80%, respectively. Three radionuclide voiding patterns were detected in children with VUR: (1) a normal voiding pattern characterised by normal FBC with near-normal PFR and EF values, (2) a markedly reduced FBC with significantly reduced VV, PFR and EF values (in children with bladder instability), and (3) a higher bladder volume with a near-normal value for PFR and a slightly reduced or near-normal EF (in children with detrusor sphincter dyssynergia). This study confirmed the association between lower urinary tract dysfunction and congenital VUR. IRNC was found to be a simple, non invasive method which allows reliable differentiation of voiding patterns in patients with VUR. IRNC can be used as a first-line method for screening in children with VUR to identify those with abnormal voiding patterns. PMID- 12536246 TI - Sestamibi is a substrate for MDR1 and MDR2 P-glycoprotein genes. AB - Technetium-99m sestamibi has attracted interest for assessment of the function of P-glycoproteins, which are well expressed in the liver and have roles in biliary transport and the removal of chemotherapeutic drugs. To further examine the cross reactivity of (99m)Tc-sestamibi for P-glycoprotein family members, we conducted studies in animals. Hepatobiliary secretion of (99m)Tc-sestamibi was determined in normal FVB/N mice, mutant mice with specific P-glycoprotein deficiencies in the FVB/N background, normal Long-Evans Agouti (LEA) rats, and Long-Evans Cinnamon (LEC) rats with abnormal copper transport and liver disease but intact P glycoprotein expression. After intrasplenic injection, (99m)Tc-sestamibi was rapidly incorporated in the mouse and rat liver, with maximal accumulation after 102+/-31 and 109+/-16 s, respectively ( P=NS). In normal mice and rats, 55%+/-11% and 55%+/-6%, respectively, of the maximal sestamibi activity was retained in the liver after 1 h ( P=NS). In double knockout mice lacking both mdr1a and mdr1b homologs of the human MDR1 ( ABCB1) gene, 88%+/-11% of maximal sestamibi activity was retained in the liver after 1 h ( P<0.001). In knockout mice deficient in either mdr1a gene or mdr2 ( ABCB4) gene, biliary sestamibi excretion was also impaired, although this impairment was relatively less pronounced in ABCB4 deficient mice than in double knockout mice lacking both ABCB1 gene homologs ( P<0.03). Hepatobiliary sestamibi excretion in LEC rats was not different from that in control normal rats, despite the presence of significant liver disease in the former. Hepatobiliary sestamibi excretion requires P-glycoproteins and is unperturbed in chronic liver disease. Sestamibi appears to be a substrate for both ABCB1 and ABCB4 genes, although the former utilizes it far more efficiently. Assessment of P-glycoprotein activity with sestamibi should consider how regulation of ABCB1 and related family members might modulate sestamibi incorporation. PMID- 12536247 TI - Is correction for age necessary in neuroimaging studies of the central serotonin transporter? AB - The central neurotransmitter serotonin plays a major role in a number of neuropsychiatric disorders. However, results from post-mortem and in vivo neuroimaging studies of the influence of age on the number of presynaptic serotonin transporters (SERT) are controversial. To provide further evidence of relevance for this ongoing discussion, SERT were imaged in 22 persons without neuropsychiatric impairment using [(123)I]2beta-carbomethoxy-3beta (iodophenyl)tropane ([(123)I]beta-CIT) and high-resolution single-photon emission tomography (SPET). The SPET analysis method was based on magnetic resonance image co-registration in order to quantify SERT with acceptable inter- and intra observer variability. Using this technique, we found a significant age-related decline in the thalamic/hypothalamic and midbrain/upper brainstem SERT density, which was more pronounced than that previously reported in the literature. From these results we conclude that age-related changes in SERT density need to be considered in future clinical studies. PMID- 12536248 TI - Sterols in microorganisms. AB - Sterols are vital components of all eukaryotic cells. This review describes the variety of sterol structures found in microalgae, yeasts, fungi, protozoans and microheterotrophs. Reports of the occurrence of sterols in prokaryotic cells are critically assessed. Methylotrophic bacteria contain unusual 4-methylsterols, but reports of 4-desmethyl sterols in cyanobacteria and other bacteria are limited and many of these seem dubious. Possible application areas for sterols derived from mass culture of microalgae and other microorganisms are highlighted. PMID- 12536249 TI - Biotechnological applications of penicillin acylases: state-of-the-art. AB - This review describes the most recent developments in the biotechnological applications of penicillin acylases. This group of enzymes is involved mainly in the industrial production of 6-aminopenicillanic acid and the synthesis of semisynthetic beta-lactam antibiotics. In addition, penicillin acylases can also be employed in other useful biotransformations, such as peptide synthesis and the resolution of racemic mixtures of chiral compounds. Particular emphasis is placed on advances in detection of new enzyme specificities towards other natural penicillins, enzyme immobilization, and optimization of enzyme-catalyzed hydrolysis and synthesis in the presence of organic solvents. PMID- 12536250 TI - Regulation of gene expression in industrial fungi: Trichoderma. AB - The genus Trichoderma comprises a group of filamentous ascomycetes that are now widely used in industrial applications because of their ability to produce extracellular hydrolases in large amounts. In addition, strong inducible promoters together with high secretory capacity have made Trichoderma an attractive host for heterologous protein production. Several promoters of genes encoding hydrolytic enzymes have been investigated in detail regarding their cis acting elements and trans-acting factors. Potent inducer molecules, for both xylanolytic and cellulolytic enzyme systems, have been identified and characterized. Furthermore, models for the recognition of the insoluble substrates cellulose and xylan have been developed based on a large set of experiments. This mini-review summarises the considerable amount of data accumulated over the past three decades. PMID- 12536251 TI - Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems. AB - In response to the rapidly growing field of proteomics, the use of recombinant proteins has increased greatly in recent years. Recombinant hybrids containing a polypeptide fusion partner, termed affinity tag, to facilitate the purification of the target polypeptides are widely used. Many different proteins, domains, or peptides can be fused with the target protein. The advantages of using fusion proteins to facilitate purification and detection of recombinant proteins are well-recognized. Nevertheless, it is difficult to choose the right purification system for a specific protein of interest. This review gives an overview of the most frequently used and interesting systems: Arg-tag, calmodulin-binding peptide, cellulose-binding domain, DsbA, c-myc-tag, glutathione S-transferase, FLAG-tag, HAT-tag, His-tag, maltose-binding protein, NusA, S-tag, SBP-tag, Strep tag, and thioredoxin. PMID- 12536252 TI - Optimization of isonovalal production from alpha-pinene oxide using permeabilized cells of Pseudomonas rhodesiae CIP 107491. AB - Optimization studies on the synthesis of isonovalal from alpha-pinene oxide by Pseudomonas rhodesiae CIP 107491 operated in a biphasic medium are presented. Three key parameters are identified. The first is the need for a permeabilization of cells by freezing them and then treating the thawed material with an organic solvent such as chloroform, toluene or diethyl ether. This operation allows both enzyme release into the aqueous phase outside the cells and an improvement in the transport properties of both substrate and product across the cell membrane, strongly increasing reaction rates. The second is that the enzyme alpha-pinene oxide lyase, which exhibits an irreversible inactivation by isonovalal (or a by product), presents a constant turn-over, i.e., the total product synthesis is proportional to the biomass loading and is close to 108 mmol (16.4 g) isonovalal l(-1) g(-1) biomass. The third phenomenon is that the biphasic system used is not phase-transfer-limited, a feature attributed to the spontaneous formation of an oil-in-water emulsion. It is thus possible to carry out a very efficient process, allowing the recovery of 2.63 mol isonovalal l(-1) (400 g l(-1)) from 25 g biomass l(-1) in 2.5 h, corresponding to an average reaction rate as high as 0.70 mmol min(-1) g(-1) cells (160 g l(-1) h(-1)). PMID- 12536253 TI - Effect of major nutrients on podophyllotoxin production in Podophyllum hexandrum suspension cultures. AB - The effect of major medium ingredients (sugar, nitrogen source and phosphate) in Podophyllum hexandrum suspension cultures was investigated in order to increase the production of podophyllotoxin, the raw material in the synthesis of anticancer drugs. Amongst B5, Eriksson, MS, Nitsch, Street and White's medium, MS medium resulted in high growth and podophyllotoxin accumulation. The optimum level of nitrogen was found to be 60 mM, with a combination of ammonium salts and nitrate in the ratio of 1:2. The highest level of podophyllotoxin was obtained at 60 g glucose/l and at 1.25 mM phosphate after 30 days. Statistical design was adopted to determine the optimum levels of the parameters for cell growth and podophyllotoxin production. PMID- 12536254 TI - Metabolic flux redistribution in Corynebacterium glutamicum in response to osmotic stress. AB - Osmotic stress constitutes a major bacterial stress factor in the soil and during industrial fermentation. In this paper, we quantified the metabolic response, in terms of metabolic flux redistribution, of a lysine-overproducing strain of Corynebacterium glutamicum grown under continuous culture, to gradually increasing osmolality. Oxygen and carbon dioxide evolution rates, and the changes in concentration of extracellular, as well as intracellular, metabolites were measured throughout the osmotic gradient. The metabolic fluxes were estimated from these measurements and from the mass balance constraints at each metabolite node of the assumed metabolic reaction network. Our results show that formation rates of compatible solutes--trehalose first and proline at a later stage of the gradient--increased with osmotic stress to equilibrate the external osmotic pressure. Estimated flux distributions indicate that the observed increase in the glucose specific uptake rate with osmotic stress is channeled through the main energy generating pathways-- glycolysis and the tricarboxylic acid cycle--while the flux through the pentose phosphate pathway remains constant throughout the gradient. This results in a significant increase in the net specific ATP production rate, which may possibly be used to support the higher energy requirements required for cellular maintenance at high osmolalities. Finally, nodal analysis confirmed that the PEP/pyruvate node is essentially rigid and that the glucose-6-phosphate, oxaloacetate and alpha-ketoglutarate nodes are flexible and therefore adaptable to changes in osmotic pressure in C. glutamicum. PMID- 12536255 TI - Development of a low-cost technology for mass production of the free-living nematode Panagrellus redivivus as an alternative live food for first feeding fish larvae. AB - The free-living nematode Panagrellus redivivus is a suitable food source for first feeding fish. In the present report, a new method for the mass production of P. redivivus is presented. The technique involves multiplication of the nematode in monoxenic (single microorganism: Saccharomyces cerevisiae) solid culture (fluid media supported by 1- to 4-cm(3) sponge cubes) in autoclavable plastic bags (size range: 50 x 30 cm to 75 x 67 cm). Two growing media were tested: oat-meal medium (OM), which is an oat-based medium (16.7% oat-meal flour in 0.8% saline solution), and purified ingredient medium (PIM), a semi-synthetic medium (1.64% meat peptone, 0.94% yeast extract, 12.6% corn starch, 0.24% glucose, 1.48% sunflower oil, in 0.8% saline solution). The bags were inoculated with 350 nematodes/g medium. After an average period of 12 days (11-13 days) at 25 degrees C, the average yield (number of nematodes/g medium) was 241 x 10(3) for OM and 333 x 10(3) for PIM in 12-l bags (50 x 30 cm). The production scale has currently reached a bag volume of 50 l (75 x 67 cm); using PIM and the conditions described above, it was possible to harvest more than 1.3 x 10(9) nematodes/bag (291 x 10(3) nematodes/g medium). In PIM, when sun flower oil was replaced with the same amount of fish oil or cod liver oil, yields of 259 x 10(3) and 290 x 10(3) nematodes/g medium, respectively, were attained. The technology for mass production and formulation of P. redivivus should enable fish-hatchery operators to rely on a cheap, standardised, and permanently available live food product for first feeding fish larvae. PMID- 12536256 TI - Increasing ethanol productivity during xylose fermentation by cell recycling of recombinant Saccharomyces cerevisiae. AB - The influence of cell recycling of xylose-fermenting Saccharomyces cerevisiae TMB3001 was investigated during continuous cultivation on a xylose-glucose mixture. By using cell recycling at the dilution rate ( D) of 0.05 h(-1), the cell-mass concentration could be increased from 2.2 g l(-1) to 22 g l(-1). Consequently, the volumetric ethanol productivity increased ten-fold, from 0.5 g l(-1) h(-1) to 5.35 g l(-1) h(-1). By increasing the biomass concentration, the xylose consumption rate increased from 0.75 g xylose l(-1) h(-1) without recycling to 1.9 g l(-1) h(-1) with recycling. The specific ethanol productivity was in the range of 0.23-0.26 g g(-1) h(-1) with or without cell recycling, showing that an increased cell-mass concentration did not influence the efficiency of the yeast. PMID- 12536257 TI - Food-grade host/vector expression system for Lactobacillus casei based on complementation of plasmid-associated phospho-beta-galactosidase gene lacG. AB - A new food-grade host/vector system for Lactobacillus casei based on lactose selection was constructed. The wild-type non-starter host Lb. casei strain E utilizes lactose via a plasmid-encoded phosphotransferase system. For food-grade cloning, a stable lactose-deficient mutant was constructed by deleting a 141-bp fragment from the phospho-beta-galactosidase gene lacG via gene replacement. The deletion resulted in an inactive phospho-beta-galactosidase enzyme with an internal in-frame deletion of 47 amino acids. A complementation plasmid was constructed containing a replicon from Lactococcus lactis, the lacG gene from Lb. casei, and the constitutive promoter of pepR for lacG expression from Lb. rhamnosus. The expression of the lacG gene from the resulting food-grade plasmid pLEB600 restored the ability of the lactose-negative mutant strain to grow on lactose to the wild-type level. The vector pLEB600 was used for expression of the proline iminopeptidase gene pepI from Lb. helveticus in Lb. casei. The results show that the food-grade expression system reported in this paper can be used for expression of foreign genes in Lb. casei. PMID- 12536258 TI - Expression of bacterial poly(3-hydroxybutyrate) synthesis genes in hairy roots of sugar beet (Beta vulgaris L.). AB - Three genes from Ralstonia eutropha necessary for poly(3-hydroxybutyrate) (PHB) synthesis were introduced into the hairy roots of sugar beet. Transformation of a vector construct harbouring the PHB genes, each fused to the coding region of the pea ribulose-bisphosphate carboxylase plastid targeting sequence, resulted in 20 transgenic hairy-root clones, producing up to 55 mg high molecular PHB/g dry weight, as identified by gas chromatography, gel permeation chromatography and HPLC. Accumulation of PHB polymer in sugar beet root leucoplasts was confirmed by transmission electron microscopy. Thus, for the first time, plastidic PHB production was demonstrated for roots of a carbohydrate-storing crop plant. PMID- 12536259 TI - Construction of a sodA::luxCDABE fusion Escherichia coli: comparison with a katG fusion strain through their responses to oxidative stresses. AB - A recombinant bioluminescent Escherichia coli strain, EBHJ, (sodA::luxCDABE), containing the promoter for the manganese superoxide dismutase ( sodA) gene fused to the Vibrio fischeri luxCDABE operon, was successfully constructed and characterized. Redox-cycling agents, such as paraquat and chromium, strongly induced a sodA- regulated response in dose-dependent manners, resulting in an increase of the bioluminescence. In a comparison with an existing oxidative stress responsive strain, DPD2511 (katG::luxCDABE), which is sensitive to H(2)O(2), the mechanism of chemicals that cause oxidative damage was elucidated via the key transcriptional factors involved in induction of the sodA and katG promoters, i.e. SoxRS and OxyR, respectively. It was found that responses from the katG- and sodA-based strains were significantly different dependent upon the chemicals being tested. Therefore, EBHJ, alone or in parallel with DPD2511, can be used to characterize and monitor chemicals that cause oxidative damage. PMID- 12536260 TI - Isolation of a novel thermophilic fungus Chaetomium sp. nov. MS-017 and description of its palm-oil mill fiber-decomposing properties. AB - Palm-oil mill fiber (POMF) is a fibrous, natural hard material discharged in enormous amounts from palm-oil mills in tropical plantations; therefore, research to find microorganisms that decompose POMF was conducted. As the result of screening, a new thermophilic fungus, Chaetomium sp. nov. MS-017, exhibiting rapid growth on POMF was isolated from rotted wood. Based on partial characterization of the decomposition of POMF, it was shown that MS-017 preferentially assimilates polysaccharides, especially hemicelluloses such as xylan. A preliminary composting study indicated that MS-017 produced 855 g of decomposed product from 1,000 g of intact POMF in 12 days under optimized solid culture conditions. The decomposition rate of POMF was 23% (w/w), and the cell yield calculated from consumed POMF was as high as 36% (w/w). These results indicate that MS-017 has a very high potential to decompose POMF and that it is suitable for economical production of compost to recycle by-product biomass from oil-palm plantations. PMID- 12536261 TI - Characteristics of a bioflocculant produced by Bacillus mucilaginosus and its use in starch wastewater treatment. AB - A bioflocculant, MBFA9, was produced from a strain of bioflocculant-producing bacteria isolated from a soil sample and identified as Bacillus mucilaginosus. MBFA9 had a good flocculating capability and could achieve a flocculating rate of 99.6% for kaolin suspension at a dosage of only 0.1 ml/l. The major component of MBFA9 was found to be polysaccharide composed mainly of uronic acid (19.1%), neutral sugar (47.4%) and amino sugar (2.7%). Infrared spectrum analysis showed the presence of carboxyl and hydroxyl groups in the bioflocculant. MBFA9 is nontoxic and can be used in food industries for suspended solids (SS) recovery. When applied to starch wastewater treatment, MBFA9 greatly accelerated the formation of flocs and the settling of organic particles in the presence of Ca(2+) salt. After 5 min of settling, the removal rate of SS and chemical oxygen demand were up to 85.5% and 68.5%, respectively, which is better than traditional chemical flocculants. PMID- 12536262 TI - Functional and structural analyses of trichloroethylene-degrading bacterial communities under different phenol-feeding conditions: laboratory experiments. AB - The effects of different phenol-feeding conditions on trichloroethylene (TCE) biodegradation and bacterial population structure in an aquifer soil community were studied. The soil sample, minerals, phenol, and TCE were mixed in glass bottles, which were then incubated under three different phenol-feeding conditions. First, phenol was supplied only once at 0.2 mM (condition 0.2P); second, it was added at 2.0 mM (condition 2.0P); and third, it was periodically supplied ten times at 0.2 mM (condition 0.2PS). TCE concentrations remained stable under conditions 0.2P and 2.0P. In contrast, TCE was completely degraded under condition 0.2PS. TCE/phenol-degrading bacteria were enumerated indirectly and functionally by quantitative PCR. The low- K(s) (half saturation constant) group of phenol-degrading bacteria, exhibiting high TCE-degrading activity, yielded a 50-fold higher population under condition 0.2PS than under condition 2.0P. The bacterial community structure under condition 0.2PS was studied by denaturing gradient gel electrophoresis targeting the genes encoding 16S rRNA and the largest subunit of multicomponent phenol hydroxylase. Sequence analysis of the major bands detected indicated the predominance of the low- K(s) group of TCE/phenol-degrading bacteria belonging to beta-Proteobacteria. These results suggest that continuous supplementation with phenol at a low concentration increases the population of the low- K(s) group of TCE/phenol-degrading bacteria. PMID- 12536263 TI - Effects on sleep architecture of pindolol, paroxetine and their combination in healthy volunteers. AB - RATIONALE: The combination of pindolol with a serotonergic antidepressant has been used to speed up the antidepressant response and to augment in cases of resistant depression. Animal studies have suggested that this increased response occurs because of 5HT(1A) antagonist properties of pindolol, which in combination with a serotonergic antidepressant produces a synergistic increase in 5HT in the synapse. OBJECTIVES: To test whether the combination of pindolol with a serotonergic antidepressant produces a synergistic increase in synaptic 5HT by examining the effects on measures of sleep, psychomotor performance and ratings of anxiety. METHODS: Twelve healthy male volunteers took part in randomised crossover study in which they received paroxetine 20 mg/day (or its placebo) for 9 days with a washout period of 5 days between. On day 7 and 9 of each treatment they also received pindolol 2.5 mg (or its placebo) three times a day. Sleep EEG recordings were made on each of the nights on pindolol (or its placebo) and ratings of saccadic eye movement parameters, subjective sleep, anxiety and other adverse events recorded on the following days. Four drug conditions were therefore tested: placebo, pindolol alone, paroxetine alone and paroxetine+pindolol. RESULTS. The combination of paroxetine+pindolol produced an increase in REM suppression and a reduction in SWS compared with other drug combinations. There were no significant effects on the other measures of 5HT function recorded in this study. CONCLUSIONS: REM suppression by the combination was approximately equal to the sum of REM suppression by each drug individually and thus does not show a synergistic effect. However, there was a significant reduction in SWS produced by only the combination treatment, which may suggest a specific effect of the combination on non-REM sleep mechanisms. PMID- 12536264 TI - Nicotine increases alcohol self-administration and reinstates alcohol seeking in rats. AB - RATIONALE AND OBJECTIVE: Alcohol and tobacco are often co-abused in humans and previous studies found that nicotine increases alcohol consumption in rats. Here, we studied whether nicotine would reinstate alcohol-taking behavior in drug-free rats and whether this effect would be enhanced by prior exposure to nicotine during alcohol self-administration training. METHODS: Rats were trained to press a lever for alcohol (12% w/v, 1 h/day), and following stable alcohol intake groups of rats ( n=11-12) were given daily vehicle or nicotine (0.2, 0.4 or 0.8 mg/kg, SC) injections just prior to the self-administration sessions for 10 days. Rats were then given 6 days of alcohol self-administration in the absence of nicotine and an additional 5-10 drug-free days during which lever presses were not reinforced (extinction). Subsequently, rats were tested for reinstatement of alcohol seeking following exposure to priming injections of vehicle or nicotine (0.4 mg/kg, SC). RESULTS: Nicotine increased alcohol self-administration in a dose- and time-dependent manner over the 10-day period. Nicotine also reinstated alcohol seeking after extinction of the alcohol-reinforced behavior, and this effect was strongly enhanced by prior nicotine exposure. CONCLUSIONS: The present data extend previous studies on the effect of nicotine on alcohol self administration, and further indicate that nicotine is an effective stimulus for reinstatement of alcohol seeking during drug-free periods. PMID- 12536265 TI - Effects of second generation of histamine H1 antagonists, cetirizine and ebastine, on the antitussive and rewarding effects of dihydrocodeine in mice. AB - RATIONALE: Little information is available about the interaction between dihydrocodeine and second-generation antihistamine drugs such as cetirizine and ebastine, with particular reference to the rewarding effect of dihydrocodeine. OBJECTIVE: The effects of second generation histamine H(1) antagonists, such as cetirizine and ebastine on the antitussive and rewarding effect of dihydrocodeine were examined in mice. METHODS: Mice were exposed to a nebulized solution of capsaicin (30 micromol/l) under conscious and identical conditions, using a body plethysmograph. The coughs produced during a 3-min exposure period were counted. Effects of H(1) antagonists on the reinforcing effect of dihydrocodeine were assessed by using the conditioned place preference procedure in mice. RESULTS: The antitussive effect of dihydrocodeine was enhanced by the simultaneous administration of either cetirizine or ebastine. There was no statistical difference between the ED(50) of dihydrocodeine in combination with ebastine and that of dihydrocodeine in combination with cetirizine. Concurrent dosing of dihydrocodeine and ebastine produced a significant place preference. This behavioral potentiation was antagonized by SCH23390, a dopamine D(1) antagonist. Moreover, ebastine enhanced the central dopamine turnover ratio, but cetirizine could not, in this study. CONCLUSION: Taken together, the potentiation of place preference of dihydrocodeine with ebastine may be due, at least in part, to stimulation of the central dopaminergic system via D(1) receptors. However, combination of dihydrocodeine with cetirizine does not potentiate place preference at all, nor does it potentiate the central dopaminergic system. Thus, it is likely that cetirizine may be a useful constituent in opioid-containing, antitussive preparations that would not potentiate the development of psychological dependence. PMID- 12536266 TI - Methemoglobin formation in children with congenital heart disease treated with inhaled nitric oxide after cardiac surgery. AB - OBJECTIVE: Inhaled nitric oxide (NO) is used as a therapy of pulmonary hypertension in children after cardiac surgery. Hemoglobin binds to NO with great affinity and forms methemoglobin by oxidation in the erythrocyte. Once produced, methemoglobin is unable to transport and unload oxygen in the tissues. The amount of available hemoglobin in the body for oxygen transport is thereby reduced. Anemia, acidosis, respiratory compromise and cardiac disease may render patients more susceptible than expected for a given methemoglobin level. The goal of the present study was to review the cumulative effect of inhaled NO on methemoglobin formation in critically ill children. We therefore looked for methemoglobin levels in children with congenital heart disease after cardiac surgery who were treated with inhaled NO in a range of 5-40 ppm. METHODS: We retrospectively reviewed the medical charts of 38 children with congenital heart disease after cardiac surgery. We extracted demographic data and physiological measurements at the following time points: (1) T0 = before starting inhaled NO therapy, (2) T1 = 24 h after the beginning of inhaled NO therapy, (3) T2 = half-time therapy, (4) T3 = end of therapy, (5) T4 = 24 h after finishing inhaled NO therapy. RESULTS: The median duration of inhaled NO therapy was 5.5 days (interquartile range 6, range 2-29), NO concentrations at T1 and T2 were 16 ppm (10, 5-40) and 12.5 ppm (12.3, 2-40), respectively. The median cumulative dose of inhaled NO was 1699 ppm (2313, 193-7018). Methemoglobin levels increased moderately, but significantly, during therapy ( T0 vs T1 p<0.05 and T0 vs T2 p<0.001). The highest methemoglobin level measured was 3.9%. Methemoglobin levels correlated positively with the inhaled NO doses applied at T1 ( r(2)=0.8376; p<0.01) and at T2 ( r(2)=0.8945; p<0.01). At T1 the methemoglobin level correlated negatively with the T1 blood pH value. The overall mortality rate was 13.2% (5 of 38 study patients died). There was no significant difference in methemoglobin levels between survivors and non survivors. CONCLUSION: We conclude from our data that the use of inhaled NO therapy for children with congenital heart disease after cardiac surgery in the described range of 5-40 ppm, resulting in a maximum of 4% methemoglobin blood level, is feasible and safe. However, we recommend the use of the minimal effective dose of inhaled NO and continuous monitoring of methemoglobin levels, especially in cases of anemia or sepsis in critically ill children. PMID- 12536267 TI - Reproducibility of the blood flow index as noninvasive, bedside estimation of cerebral blood flow. AB - OBJECTIVE: To investigate the feasibility and reproducibility of the blood flow index (BFI) method for measuring cerebral blood flow. DESIGN AND SETTING: Prospective functional study in pediatric intensive care. PATIENTS AND PARTICIPANTS: 14 consecutive patients with median age of 2 months (range 1 days 11 years) requiring artificial ventilation, invasive arterial blood pressure monitoring, and central venous access. INTERVENTIONS: The first passage of an intravenous indocyanine green (ICG) bolus through the cerebral vasculature was monitored by noninvasive near-infrared spectroscopy. BFI was calculated by dividing maximal ICG absorption change by rise time. Reproducibility was evaluated by six ICG injections at 5-min intervals. RESULTS: Of all ICG injections 6% were canceled, and 4% were eliminated due to injection failures. Median BFI of 17 reproducibility determinations was 71 (range 12-213) and median coefficient of variation (CV) of BFI was 10% (4.9-18.5). The quantity of ICG bolus did not affect the CV (0.1 vs. 0.3 mg ICG/kg). Eight reproducibility tests in patients after cardiac surgery had smaller CV than the others, and the eight in newborns had higher CV than in older children. Patient parameters such as arterial blood pressure, endtidal CO(2), and percutaneous oxygen saturation were stable and showed CV below 2% during reproducibility determination. CONCLUSIONS: The BFI method allows rapid and repeated measurements of CBF with good feasibility and reproducibility. As a relative but not absolute measure of CBF, BFI seems to be suited for clinical evaluation of intraindividual CBF changes during determination of cerebrovascular reactivities or during therapeutic interventions. PMID- 12536268 TI - Fluid responsiveness in mechanically ventilated patients: a review of indices used in intensive care. AB - OBJECTIVE: In mechanically ventilated patients the indices which assess preload are used with increasing frequency to predict the hemodynamic response to volume expansion. We discuss the clinical utility and accuracy of some indices which were tested as bedside indicators of preload reserve and fluid responsiveness in hypotensive patients under positive pressure ventilation. RESULTS AND CONCLUSIONS: Although preload assessment can be obtained with fair accuracy, the clinical utility of volume responsiveness-guided fluid therapy still needs to be demonstrated. Indeed, it is still not clear whether any form of monitoring-guided fluid therapy improves survival. PMID- 12536269 TI - Surviving intensive care: a report from the 2002 Brussels Roundtable. AB - The traditional goal of intensive care has been to decrease short-term mortality. While worthy, this goal fails to address the issue of what it means to survive intensive care. Key questions include whether intensive care survivors have optimal long-term outcomes and whether ICU care decisions would change if we knew more about these outcomes. The 2002 Brussels Roundtable, "Surviving Intensive care", highlighted these issues, summarizing the available evidence on natural history and risk factors for critical illness and outlining future directions for care and research. Critical illness is associated with a wide array of serious and concerning long-term sequelae that interfere with optimal patient-centered outcomes. Although traditional short-term outcomes, such as hospital mortality, remain extremely important, they are not likely to be adequate surrogates for subsequent patient-centered outcomes. As such, it is important to focus specifically on how critical illness and intensive care affects a patient's and relatives' long-term health and well-being. There are a large number of potential pre-, intra-, and post-ICU factors that may improve or worsen these outcomes, and these factors are subjects for future research. In addition, future clinical trials of ICU therapies should include long-term follow-up of survival, quality of life, morbidity, functional status, and costs of care. Follow-up ought to be for at least six months. The SF-36 and EuroQOL EQ-5D are the best-suited instruments for measuring quality of life in multicenter critical care trials though further methodologic research and instrument design is encouraged. There are also opportunities today to improve care. Key to taking advantage of such opportunities is the need for a global awareness of critical illness as an entity that begins and ends outside the ICU 'box'. Specific interventions that show promise for improving care include ICU discharge screening tools and ICU follow up clinics. PMID- 12536270 TI - Puumala virus infection with multiorgan involvement. PMID- 12536271 TI - External validation of the SAPS II, APACHE II and APACHE III prognostic models in South England: a multicentre study. AB - OBJECTIVE: External validation of three prognostic models in adult intensive care patients in South England. DESIGN. Prospective cohort study. SETTING: Seventeen intensive care units (ICU) in the South West Thames Region in South England. PATIENTS AND PARTICIPANTS: Data of 16646 patients were analysed. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: We compared directly the predictive accuracy of three prognostic models (SAPS II, APACHE II and III), using formal tests of calibration and discrimination. The external validation showed a similar pattern for all three models tested: good discrimination, but imperfect calibration. The areas under the receiver operating characteristics (ROC) curves, used to test discrimination, were 0.835 and 0.867 for APACHE II and III, and 0.852 for the SAPS II model. Model calibration was assessed by Lemeshow-Hosmer C-statistics and was Chi(2 )=232.1 for APACHE II, Chi(2 )=443.3 for APACHE III and Chi(2 )=287.5 for SAPS II. CONCLUSIONS: Disparity in case mix, a higher prevalence of outcome events and important unmeasured patient mix factors are possible sources for the decay of the models' predictive accuracy in our population. The lack of generalisability of standard prognostic models requires their validation and re calibration before they can be applied with confidence to new populations. Customisation of existing models may become an important strategy to obtain authentic information on disease severity, which is a prerequisite for reliably measuring and comparing the quality and cost of intensive care. PMID- 12536273 TI - Should the autopsy be resuscitated? PMID- 12536272 TI - Lung computed tomography during a lung recruitment maneuver in patients with acute lung injury. AB - OBJECTIVE: To assess the acute effect of a lung recruitment maneuver (LRM) on lung morphology in patients with acute lung injury (ALI) or acute respiratory distress syndrome (ARDS). PATIENTS: Ten patients with ALI/ARDS on mechanical ventilation. DESIGN: Prospective clinical study. SETTING: Computed tomography (CT) scan facility in a teaching hospital. INTERVENTIONS: An LRM performed by stepwise increases in positive end-expiratory pressure (PEEP) of up to 30-40 cm H(2)O. Lung basal CT sections were taken at end-expiration (patients 1 to 5), and at end-expiration and end-inspiration (patients 6 to 10). Arterial blood gases and static compliance (C(st)) were measured before, during and after the LRM. MEASUREMENTS AND MAIN RESULTS: Poorly aerated and non-aerated tissue at PEEP 10 cm H(2)O accounted for 60.0+/-29.1% of lung parenchyma, while only 1.1+/-1.8% was hyperinflated. Increasing PEEP to 20 and 30 cm H(2)O, compared to PEEP 10 cm H(2)O, decreased poorly aerated and non-aerated tissue by 16.2+/-28.0% and 33.4+/ 13.8%, respectively ( p<0.05). This was associated with an increase in PaO(2) and a decrease in total static compliance. Inspiration increased alveolar recruitment at all PEEP levels. Hyperinflated tissue increased up to 2.9+/-4.0% with PEEP 30 cm H(2)O, and to a lesser degree with inspiration. No barotrauma or severe hypotension occurred. CONCLUSIONS: Lung recruitment maneuvers improve oxygenation by expanding collapsed alveoli without inducing too much hyperinflation in ALI/ARDS patients. An LRM during the CT scan gives morphologic and functional information that could be useful in setting ventilatory parameters. PMID- 12536274 TI - Animal cooperation among unrelated individuals. AB - The evolution of cooperation has long been a topic near and dear to the hearts of behavioral and evolutionary ecologists. Cooperative behaviors run the gamut from fairly simple to very complicated and there are a myriad of ways to study cooperation. Here I shall focus on three paths that have been delineated in the study of intraspecific cooperation among unrelated individuals: reciprocity, byproduct mutualism, and group selection. In each case, I attempt to delineate the theory underlying each of these paths and then provide examples from the empirical literature. In addition, I shall briefly touch upon some recent work that has attempted to examine (or re-examine) the role of cognition and phylogeny in the study of cooperative behavior. While empirical and theoretical work has made significant strides in the name of better understanding the evolution and maintenance of cooperative behavior in animals, much work remains for the future. "From the point of view of the moralist, the animal world is on about the same level as the gladiator's show. The creatures are fairly well treated, and set to fight; whereby the strongest, the swiftest and the cunningest live to fight another day. The spectator has no need to turn his thumb down, as no quarter is given em leader the weakest and the stupidest went to the wall, while the toughest and the shrewdest, those who were best fitted to cope with their circumstances, but not the best in any other way, survived. Life was a continuous free fight, and em leader a war of each against all was the normal state of existence." (Huxley 1888) PMID- 12536275 TI - Chemical interactions between amino acid and RNA: multiplicity of the levels of specificity explains origin of the genetic code. AB - The emergence of the genetic code remains an enigma. Proposed mechanisms are based on random, historical, thermodynamic and natural selection. However, they introduce chance as a key factor for overcoming the difficulties encountered by the model. We propose here a model in which three successive levels of chemical specificity generated the nucleotide assignments of amino acids in the genetic code. The first level results from hydrophobic and stereospecific interactions between amino acids and short oligonucleotides (termed oligons). The second and third levels of specificity are determined by conditions of energy transfer from loaded oligons (amino acid-oligomer covalently linked) to formation of phosphodiester bond (second level of specificity) and peptidic bond (third level of specificity), while these reactions are catalyzed by RNA templates. This model is sustained by the relationships observed between dipole moments of the nucleotides (forming the anticodon) and reactivity of the amino acyl linkage of the loaded oligon. Moreover, analysis of modern tRNAs reveals that they were probably generated by loose duplication of the nucleotide sequence forming the oligons, after emergence of the 'genetic code.' Indeed, the similarity of nucleotide composition with that of the anticodon decreases with the tRNA domain's distance from the anticodon, but the acceptor stem is relatively more similar to the anticodon than other stems closer to it. This would be because energy transfer constraints that existed between anticodon and amino acid in prebiotic loaded oligonucleotides still affect the structures of modern tRNA acceptor stems. In the model presented, the genetic code is inherent to the most archaic 'molecular physiology' in protolife, even before emergence of a functional 'protein world.' Simple physical processes, in which a level of specificity is integrated in an emerging meta-structure expressing new properties, generate a parsimonious and realistic explanation of emergence of the genetic code. PMID- 12536276 TI - Adult flight in a water strider exposed to habitat drydown in the larval stages. AB - Aquarius paludum inhabits a wide variety of water surfaces, ranging from temporary to permanent ponds and lakes throughout the Palearctic. Thus, some larvae of A. paludum will be exposed to habitats that dry out. We assess here the influence of drying on water strider growth and development. When exposed to drying-out in the larval stage, the growth period was prolonged. Drying-out during one of the first three instars was associated with longer maintenance of flight muscles without histolysis and reduced the number of eggs laid by long winged (LW) adults that shifted to a "flyer" strategy. Alternatively, adults exposed to dry habitats during whole larval period expressed shorter preoviposition periods and shifted to a "non-flying, reproducer" strategy. These contrasting responses to drying can be understood as flexible physiological functions depending on the drying history to which larvae are exposed. This flexibility may be a significant factor in allowing A. paludumto occupy such a wide range of habitats in Eurasia. PMID- 12536277 TI - Mammalian neurohormones: potential significance in reproductive physiology of St. John's wort (Hypericum perforatum L.)? AB - Melatonin and serotonin are indoleamine neurohormones that function as photoperiod signals in many species and have recently been found in St. John's wort, a medicinal plant used in the treatment of depression. There is no known role for melatonin in higher plants but melatonin functions as a signal of changes in photoperiod in other species. In the current study, serotonin and melatonin were quantified during flower development. Higher concentrations of serotonin were found in flower buds at the tetrad stage of microspore development and higher melatonin concentrations were detected during uninucleate mircosporogenesis. Additionally, the regeneration potential of isolated anthers was highest in the same stage that had elevated melatonin concentrations. These data provide the first evidence of the presence of melatonin during flower development and raise many questions about the potential roles of serotonin and melatonin as regulatory molecules in the reproductive flexibility of higher plants. PMID- 12536278 TI - Mimicry of queen Dufour's gland secretions by workers of Apis mellifera scutellata and A. m. capensis. AB - The development of the Dufour's gland of workers of the two honey bee races Apis mellifera scutellata and A. m. capensis was measured. The Dufour's glands of A. m. capensis workers were longer and increased in length more rapidly than the glands of workers of A. m. scutellata at comparable ages. Analysis of the Dufour's gland secretions of workers and queens of both races revealed that there were caste and racial differences. Secretions of queenright A. m. scutellata workers were dominated by a series of long-chain hydrocarbons. In contrast the secretions of the A. m. capensis workers both under queenright and queenless conditions were a mixture of hydrocarbons and wax-type esters, as were those of queens. Multivariate analysis of the secretion profiles indicated that laying workers of both races mimic queens. The secretions of the A. m. capensis laying workers mimicked queen secretions most closely, enabling them to act as successful social parasites. PMID- 12536279 TI - Seven suggestive quantitative trait loci influence hygienic behavior of honey bees. AB - In 1964, Walter Rothenbuhler proposed a two-gene model to explain phenotypic variance in the remarkable behavior in which honey bee workers remove dead brood from their colonies. Rothenbuhler's model proposed that one locus controls the uncapping of brood cells containing dead pupae, while a second controls the removal of the cell contents. We show here, through molecular techniques and quantitative trait loci (QTL) linkage mapping, that the genetic basis of hygienic behavior is more complex, and that many genes are likely to contribute to the behavior. In our cross, we detected seven suggestive QTLs associated with hygienic behavior. Each detected QTL controlled only 9-15% of the observed phenotypic variance in the character. PMID- 12536280 TI - Aerosols: unexpected disequilibrium phenomena between airborne radio activities of lead-210 and its progenies bismuth-210 and polonium-210. AB - For the first time, concentrations of the long lived radon progenies (210)Pb, (210)Bi and (210)Po were measured in the mine atmosphere of the so called "healing gallery" in Badgastein, Austria, a region famous for its radioactive springs. These investigations were performed in order to study the radioactive equilibrium between the (210)Pb-(210)Bi and the (210)Pb-(210)Po pairs so as to gain more information about the aerosol-forming processes in the mine. The particle size distribution of the aerosols was determined under different ventilation conditions. Six-stage and eight-stage cascade impactors with working ranges from 0.15 to 5 micro m and from 0.063 to 8 micro m, respectively, were used to collect the mine aerosols. These samples were analysed in the laboratory and measured by liquid scintillation spectrometry. The most surprising results were found under full ventilation, when the total activity concentrations of (210)Pb, (210)Bi and (210)Po were 4.6, 2.0 and 16.5 mBq/m(3), respectively. In this case (210)Po/(210)Pb activity ratios ranged between 1.8+/-0.3 and 4.3+/-0.3. These unexpected results were confirmed by the eight-stage impactor samples. For the smallest particles, between 0.062 and 0.125 micro m, an even higher value of 7.5 was observed. As outside sources could be excluded, such (210)Po enrichments must occur during the aerosol-forming process itself inside the mine. PMID- 12536281 TI - The functionality of female reciprocal calls in the Iberian midwife toad (Alytes cisternasii): female-female acoustic competition? AB - Female midwife toads (genus Alytes) emit highly variable reciprocal calls of unclear function prior to and during courtship. In some species, female-female competition, expressed as physical fighting, has been reported. Males of Majorcan midwife toads (Alytes muletensis) show phonotactic response to female calls, and females of Iberian midwife toads (Alytes cisternasii) respond differently according to the male call characteristics. In this study, I test the hypothesis of female-female acoustic competition as an additional function of female reciprocal calls. Playback tests indicate that female calls are not clearly involved in female acoustic competition in the Iberian midwife toad, therefore female calls could be directed at males rather than towards competitive females. PMID- 12536282 TI - Tooth hardness increases with zinc-content in mandibles of young adult leaf cutter ants. AB - A wide variety of arthropods and members of other phyla have elevated concentrations of Zn, Mn, other heavy metals and halogens in their jaws, leg claws, and other "tools" for interacting with the environment. While measured Zn concentrations reach 25% of dry mass in scorpion stings, concentrations are often lower than this and the enriched structures are not heavily biomineralized like vertebrate teeth and the radula of mollusks. For this reason, the degree to which the inorganic components of these structures modify their mechanical properties is in question. Here we address this problem by measuring hardness during the development of Zn accumulations in ant mandibles. We found that Zn is incorporated into the mandibular teeth of leaf-cutter ants during early adult life, reaching concentrations of about 16% of dry mass. We show that the hardness of the mandibular teeth increases nearly three-fold as the adults age and that hardness correlates with Zn content ( r=0.91). We suggest that young adults rarely cut leaves partly because their mandibles are not yet rich in Zn. Zinc enrichment (along with enrichment by other heavy metals and halogens) may play an unrecognized role in the behavioral ecology and evolution of a wide variety of invertebrates. PMID- 12536283 TI - Glutamic acid decarboxylase autoantibodies and neurological disorders. AB - Glutamic acid decarboxylase (GAD) is the enzyme that catalyses the production of GABA, a major neurotransmitter of the central nervous system. Antibodies to GAD (GAD-Ab) were first recognised in a patient affected by stiff-person syndrome; subsequently they were reported in a large number of cases with type 1 diabetes. Recently GAD-Ab have been described in a number of patients affected by chronic cerebellar ataxia, drug-resistant epilepsy and myoclonus. These cases usually harbour other autoantibodies or are affected by organ-specific autoimmune diseases. The role of GAD-Ab is still unclear; the lack of experimental models makes it difficult to investigate their potential pathogenetic role. However two mechanisms have been suggested: the reduction by GAD-Ab of GABA synthesis in nerve terminals or the interference with exocytosis of GABA. PMID- 12536284 TI - Famous face recognition and naming test: a normative study. AB - Tests of famous face recognition and naming, and tasks assessing semantic knowledge about famous people after presentation either of their faces or their names are often used in the neuropsychological examination of aphasic, amnesic and demented patients. A total of 187 normal subjects took part in this study. The aim was to collect normative data for a newly devised test including five subtests: famous face naming, fame judgement after face presentation and after name presentation, semantic knowledge about famous people after face presentation and after name presentation. Norms were calculated taking into account demographic variables such as age, sex and education and adjusted scores were used to determine inferential cut-off scores and to compute equivalent scores. Multiple regression analyses showed that age and education influenced significantly the performance on most subtests, but sex had no effect on any of them. Scores of the subtest evaluating fame judgements after name presentation were significantly influenced only by education. The only subtest whose scores were not influenced by any demographic variable was fame judgement after face presentation. PMID- 12536285 TI - General Knowledge of the World: a standardised assessment. AB - An entirely verbal enquiry of "General Knowledge of the World" made up by 168 questions exploring 14 domains of knowledge (12 questions each) has been standardised on 175 (97 women and 78 men) healthy Italian subjects with at least 8 years of formal education. Norms for each set of questions are provided. An unexpected finding is that age did not play an influential role on performance, whereas education did: the higher it was, the better the score. Women proved to fare significantly worse than men. A feasibility check on 30 Alzheimer patients with very mild overall cognitive impairment showed that the general knowledge enquiry was relatively easy to administer. The discrimination power between normal controls and Alzheimer patients for each subtest has been calculated. PMID- 12536286 TI - Continuous infusion of midazolam in the treatment of refractory generalized convulsive status epilepticus. AB - We studied the efficacy and safety of midazolam given as a continuous infusion in the treatment of refractory generalized convulsive status epilepticus (RGCSE). We carried out a prospective, open study, in 19 patients (11 men) with RGCSE in the intensive care unit at Firat Medical Center in Elazig. When intravenous administration of 0.3 mg/kg diazepam (three times at 5-min intervals), 20 mg/kg phenytoin, and 20 mg/kg phenobarbital failed to bring the episode under control, patients were administered an intravenous bolus of midazolam (200 microg/kg) followed by a continuous infusion at 1 microg/kg min. The dose was increased by 1 microg/kg min every 15 min until the episode of seizure was brought under control. The time from beginning of treatment to control of seizures, infusion rate, and side-effects were monitored. The mean age of the patients was 40.4 years (range 16-87 years). The clinical etiology of RGCSE was idiopathic epilepsy (6 cases), anoxicischemic cerebral insult due to cardiac arrest (3), viral encephalitis (2), intrahemispheric hematoma due to hemorrhagic stroke (1), cerebral infarct due to ischemic stroke (1), pituitary adenoma (1), post traumatic epilepsy (1), renal failure (1), tuberculous meningitis (1), and unknown (2). In eighteen (94.7%) patients, seizures were completely controlled in a mean time of 45 min (range, 5-120 min) at a mean infusion rate of 8 microg/kg min (range, 3-21 microg/kg min). In one patient seizures did not stop. Midazolam administration did not cause any significant change in blood pressure, heart rate, oxygen saturation, or respiratory status. The mean time to full consciousness for patients after stopping the infusion was 1.6 hours (range, 2.0 8.5 hours). The mean infusion duration of midazolam was 14.5 hours (range, 12-25 hours). Midazolam is an effective and safe drug to control RGCSE, and may represent a substantial improvement over current therapeutic approaches such as pentobarbital anesthesia. PMID- 12536287 TI - Serum uric acid and multiple sclerosis. AB - Several studies indicate that patients with multiple sclerosis (MS) have low serum levels of the endogenous antioxidant uric acid (UA), although it has not been established whether UA is primarily deficient or secondarily reduced due to its peroxynitrite scavenging activity. We measured serum urate levels in 124 MS patients and 124 age- and sex-matched controls with other neurological diseases. In addition, we compared UA levels when MS patients were stratified according to disease activity (by means of clinical examination and MRI), duration, disability and course. MS patients had significantly lower serum urate levels than controls (p= 0.001). However, UA levels did not significantly correlate with disease activity, duration, disability or course. Our study favors the view that reduced UA in MS is a primary, constitutive loss of protection against oxidative agents, which deserves further pathogenetic elucidation aimed at future therapeutic strategies. PMID- 12536288 TI - Macrophage-secreted myogenic factors: a promising tool for greatly enhancing the proliferative capacity of myoblasts in vitro and in vivo. AB - In this work we set out to determine if the murine macrophage J774 cell line can be used to produce myogenic growth factors. Activated J774 macrophages were grown in serum-free conditions. The macrophage-conditioned medium (MCM) was then used to treat cultures of primary myoblasts and regenerating muscle tissue, in vitro and in vivo respectively. MCM activity in vitro was tested by analyzing the expression of muscle-specific transcription factors, in parallel with the proliferation and differentiation rates of the cells. The macrophage-secreted factors greatly enhanced the proliferative potential of both rat and human primary myoblasts and were found to be highly muscle-specific. In vivo, MCM administration markedly enhanced the regenerative processes in damaged muscles. The ability to produce large amounts of macrophage-secreted myogenic factor(s) in the absence of serum holds great promise for its biochemical characterization and successive application in therapeutic protocols, both for ex vivo gene therapy and for muscle repair. PMID- 12536289 TI - X-linked Charcot-Marie-Tooth disease caused by a novel point mutation in the connexin-32 gene. AB - We report the clinical and electrophysiological findings of a patient with X linked Charcot-Marie-Tooth disease and a novel point mutation in the connexin-32 gene. A 31-year-old man presented with a 5 year history of progressive imbalance and distal weakness in his legs. Electrophysiological studies confirmed an asymmetric, predominantly axonal sensorimotor neuropathy with some demyelinating features. Genetic testing revealed a G/A transition (Ala40Thr) in a conserved transmembrane region of the connexin-32 gene. PMID- 12536290 TI - Subarachnoid haemorrhage with "Ecstasy" abuse in a young adult. AB - Abuse of the drugs like amphetamine, cocaine and "Ecstasy" may be complicated by intracerebral, subdural or subarachnoid haemorrhage. Contrary to historical opinion, drug-related intracranial haemorrhage (ICH) is frequently related to an underlying vascular malformation. We report the case of an 18-year-old man with a history of Ecstasy abuse preceding the onset of severe occipital headache. Cerebral computed tomography revealed right-sided subarachnoid haemorrhage and cerebral angiography showed right-sided middle cerebral artery aneurysm of 1 cm diameter. The patient was treated surgically with aneurysm clipping. Three weeks after onset of intracranial haemorrhage, neurological examination demonstrated normal findings. A history of severe headache immediately after using amphetamine, Ecstasy, or cocaine should alert doctors to the possibility of intracerebral haemorrhage. Arteriography should be part of the evaluation of most young patients with stroke or non-traumatic ICH. PMID- 12536291 TI - Pancreatic encephalopathy: a 7-year follow-up case report and review of the literature. AB - Pancreatic encephalopathy is a rare complication of acute pancreatitis. Clinical features include focal neurological signs and acute onset of dementia. This picture can fluctuate over time: cyclic progression with remission and relapses has been described. We present the case of a 43-year-old man who, after an acute episode of pancreatitis, experienced five relapses, with alternating focal signs. The patient has improved, but cognitive impairment persists after a 7-year follow up. PMID- 12536292 TI - Gene transfer by adenovirus-mimetic peptides in the presence of a cationic lipid and/or adenovirus. Analysis of the contribution of the viral and nonviral components. AB - Peptide and cationic lipid-based gene transfer vectors have shown promise for gene therapy but are still less efficient than viral gene transfer vectors. We have examined the mechanism of gene transfer of different adenovirus-mimetic peptides in the presence and absence of a cationic lipid, lipofectamine and/or adenovirus with the aim of improving the design of nonviral vectors for efficient gene transfer. Three polylysine-adenovirus-mimetic peptides were synthesised and examined for their efficacy for gene transfer. Transfection levels in four cell lines: adenovirus permissive human tracheal epithelial (56FHTE8o(-)), human lung carcinoma (A549), human colon carcinoma (Caco-2) cells, and adenovirus low permissive Chinese hamster ovary (CHO) cells, were examined. The polylysine adenovirus-mimetic peptides increased the level of transfection of a reporter transgene in all cell lines. Transfection was substantially increased when an adenovirus was added to cells after pre-incubation with the vector complexes. Formulation of the peptide vector complexes with lipofectamine increased their transfection efficacy and the subsequent addition of an adenovirus increased transfection levels even further but only in permissive cells. Pre-incubation of cells with lipofectamine-peptide vector complexes increased cell binding of the adenovirus but uptake was only increased in intermediate- or non-permissive cells. The addition of lipofectamine increased transgene expression of a recombinant adenovirus in non-permissive cells but not in permissive cells. Enhancement with an adenovirus of peptide vector gene transfer is probably due to more efficient endosome escape while enhancement of gene transfer by peptide vectors complexed to lipofectamine is due to an increase in cellular binding and/or internalisation of the adenovirus. PMID- 12536293 TI - Analysis of oropouche virus L protein amino acid sequence showed the presence of an additional conserved region that could harbour an important role for the polymerase activity. AB - We described here the complete nucleotide sequence of the L RNA segment of Oropouche virus (genus Orthobunyavirus, family Bunyaviridae). We found the L RNA segment is 6846 nucleotides long and encodes a putative RNA polymerase of 2250 amino acids. Phylogenetic analysis showed that ORO virus cluster to the Orthobunyavirus genus confirming the serological classification. It also showed that Bunyamwera and California viruses, from the Orthobunyavirus genus, are more closely related to each other than to ORO virus. Sequence comparisons performed between the L proteins of 15 bunyaviruses and the PB1 proteins of 3 influenza viruses revealed that ORO L protein contains the 3 regions characteristic of arenaviruses and bunyaviruses. These comparisons also showed the existence of an additional fourth conserved region in the L protein of bunyaviruses that contains at least two active sites. PMID- 12536294 TI - Host range of poliovirus is restricted to simians because of a rapid sequence change of the poliovirus receptor gene during evolution. AB - The host range of most poliovirus (PV) strains is restricted to simians. This host range specificity is believed to be determined by the interaction between PV and its receptor molecule. To elucidate the molecular basis of this species specific infection of PV, we cloned orthologs of the PV receptor (PVR) gene ( pvr) as well as those of PV receptor-related genes 1 and 2 ( prr1 and prr2) from various mammalian species. These three genes are widely present in mammalian genomes including those of non-susceptible species. Comparison of the deduced amino acid sequences of PVR orthologs revealed that the NH(2)-terminal immunoglobulin-like domain (domain 1), which is the virus binding site in the human PVR, is highly variable among species, whereas that of PRR1 is highly conserved. Domain 1 of the PVR orthologs for the ring-tailed lemur and rabbit, which are not susceptible to PV, show only 51 and 61% amino acid sequence identity to that of human PVR, respectively. Chimeric PVR proteins that have the domain 1 of the ring-tailed lemur and rabbit PVRs failed to serve as receptors for PV. These results suggest that rapid changes in the domain 1 sequence during mammalian evolution determined the host range restriction of PV. PMID- 12536295 TI - Identification of differentially expressed genes in brains of newborn Borna disease virus-infected rats in the absence of inflammation. AB - Infection of newborn rats with Borna disease virus (BDV) leads to viral persistence in the central nervous system without overt signs of inflammation. Nevertheless, these rats display distinct behavioral and neurodevelopmental abnormalities. The molecular basis of the latter is still unknown. Using a cDNA array representing 1200 genes, we sought to identify cellular genes which are differentially expressed following perinatal BDV-infection. RNA samples prepared from different brain regions were analysed at various time points before or after BDV-induced defects become evident. In infected brains, we found upregulated expression of genes encoding brain fatty acid binding protein (B-FABP), beta2 microglobulin (beta2m) and, as described previously, the chemokine IP-10. Kinetic studies revealed sustained increased expression of B-FABP in infected frontal cortices beginning about three weeks p.i. Moreover, a slight transient increase of B-FABP expression in infected hippocampi was observed 3-5 weeks p.i. In situ hybridization studies combined with immunohistochemistry suggested that expression of beta2m was predominantly upregulated in glial cells and possibly also in some neurons. Employing cultured infected hippocampus slices and infected genetically modified mice, we provide evidence, that the observed upregulation of beta2m expression is not triggered by IFN-gamma, but rather by IFN-alpha/beta. PMID- 12536296 TI - Phylogenetic analysis of rabbit haemorrhagic disease virus in France between 1993 and 2000, and the characterisation of RHDV antigenic variants. AB - The first molecular epidemiological study of Rabbit haemorrhagic disease virus undertaken in France between 1988 and 1995, identified three genogroups, two of which (G1, G2) disappeared quickly. We used immunocapture-RT-PCR and sequencing to analyse 104 new RHDV isolates collected between 1993 and 2000. One isolate was obtained in 2000 from a French overseas territory, the Reunion Island. The nucleotide sequences of these isolates were aligned with those of some French RHDV isolates representative of the three genogroups previously identified, of some reference strains and German and American RHDV antigenic variants. Despite the low degree of nucleotide sequence variation, three new genogroups (G4 to G6) were identified with significant bootstrap values. Two of these genogroups (G4 and G5) were related to the year in which the RHDV isolates were collected. Genogroup G4 emerged from genogroup G3, which has now disappeared. Genogroup G5 is a new independent group. The genogroup G6 contained an isolate collected in mainland France in 1999 and the isolate collected from the Reunion Island, as well as German and American RHDV variants. Multiple sequence alignments of the VP60 gene and antigenic analysis with monoclonal antibodies demonstrated that these French isolates are two new isolates of the RHDV variant. PMID- 12536297 TI - Molecular diversity of RNA-2 genome segments in pecluviruses causing peanut clump disease in West Africa and India. AB - The complete nucleotide sequence of RNA-2 genome segments of four isolates of Peanut clump virus (PCV) and two isolates of Indian peanut clump virus (IPCV) were determined. Comparisons among the complete RNA-2 sequences of six isolates from this study and two published earlier, revealed a high degree of variability in size (between 4290 and 4652 nucleotides) and nucleotide sequence identities (between 58 % and 79 %). Amino acid sequence alignments of the five open reading frames (ORF) showed that ORF 4, which encodes the second of the triple gene block proteins, is highly conserved (90 to 98 % identical) whereas the protein encoded by ORF 2, whose function is unknown, is less conserved (25 to 60 % identical). The coat protein of the eight isolates showed amino acid identities between 37 % and 89 % and contained several conserved residues. Phylogenetic comparisons, based on complete RNA-2 sequences, revealed that the eight isolates grouped into two distinct clusters with no geographical distinction between PCV and IPCV isolates. Phylogenetic tree topologies for individual ORFs showed an overall similarity with that obtained from entire RNA-2 sequences, although the relative positions of individual isolates vary within each cluster. The results indicate that there is substantial divergence among the RNA-2 genomes of pecluviruses and suggest that different proteins have evolved differently, possibly due to different selection pressures. PMID- 12536298 TI - Interaction of Bombyx mori nucleopolyhedrovirus BRO-A and host cell protein laminin. AB - The Bombyx mori nucleopolyhedrovirus (BmNPV) contains five baculovirus repeated ORF ( bro) genes, all of which are expressed as delayed early genes. We have recently reported that BmNPV BRO proteins, specially BRO-A and BRO-C, contain a nucleic acid binding activity and are involved in nucleosome structures in nuclei of infected cells. To further understand the function of bro-a gene, we looked for factors interacting with BmNPV BRO-A using the yeast two-hybrid system. Fifteen clones obtained from a cDNA library of mock-infected cells and one from a library prepared at 2 h postinfection (p.i.) were found to comprise one distinct gene, which was identified as the Bombyx homolog (bLaminin) of Drosophila laminin beta1. A direct interaction between BRO-A and N-terminal region of bLaminin was demonstrated by in vitro pull-down experiments. Further pull-down assays using BmN cell extracts and anti-laminin antibodies also showed interaction of both proteins. In addition, two more clones were obtained from cDNA library of 12 h p.i. and were found to encode BRO-A itself, indicating that BRO-A forms an oligomer. Taken together, we propose that BRO-A may function as a laminin binding protein. PMID- 12536299 TI - Genetic diversity of avian infectious bronchitis virus California variants isolated between 1988 and 2001 based on the S1 subunit of the spike glycoprotein. AB - Twenty-nine isolates of avian infectious bronchitis virus (IBV) recovered from commercial chicken flocks in California between 1988 and 2001 and identified as California variants by serotype and direct automated cycle sequencing of the IBV spike glycoprotein S1 subunit, were further characterized phylogenetically and by nucleotide sequence comparison. California variants were grouped according to production type of chicken, by comparison with public access sequence databases (NCBI GenBank and EMBL), or based on phylogenetic analysis. Fisher's Exact test was used to compare mutations per year, purifying and positive selection, predictive antigenicity, and a > or = 6 bp deletion between California variant groups.A high number of mutations at the nucleotide level ( p = 0.013) and a > or = 6 bp deletion in the nucleotide sequence ( p = 0.006) was significantly associated with broiler-type chickens. However, 88% of significant comparisons at the amino acid level such as purifying and positive selection were seen in layer type chickens. A pronounced predictive antigenicity in the HVR2 region was also associated with layer-type chickens ( p = 0.001). The study indicates that IBV in California is in a phase of slow evolution with different evolutionary patterns being associated with the production type of chicken. PMID- 12536300 TI - Mapping of the bovine herpesvirus 1 glycoprotein C promoter region and its specific transactivation by the viral BICP27 gene product. AB - We investigated the cis-acting sequences of the promoter regulating the gene encoding the glycoprotein C (gC) of bovine herpesvirus 1 (BoHV-1), a gene of the gamma1 class. S1 nuclease protection assays revealed that gC transcription initiated predominantly at position C18281 of the viral genome, corresponding to 21 and 56 bases downstream from putative TATA-like and CAAT boxes, respectively. To map the gC promoter (pgC), we measured the ability of a series of nested deletions of the region +71 to -1155 with respect to the major gC transcriptional start site, to drive expression of the firefly luciferase (Luc) reporter gene following transient transfection of a cell host, in the absence as well as in the presence of viral factors expressed in trans. We show that the minimal pgC sequences required to drive maximal BoHV-1 independent expression was within the region +71 to -76 which harbours the putative TATA and CAAT boxes, the mRNA start site, and the complete 5' transcript leader sequence. This small pgC fragment was highly trans-activated by the co-expression of the BoHV-1-encoded BICP27 protein, but not BICP0 nor BTIF. In contrast, the pgC fragment spanning the region +71 to 1155 was only minimally trans-activated by BICP27, but substantially stimulated by BICP0. These findings thus suggest that gC gene regulation may involve the combined action of several viral transactivators. PMID- 12536301 TI - Apoptosis in canine distemper. AB - Canine distemper is a systemic viral disease characterized by immunosuppression followed by secondary infections. Apoptosis is observed in several immunosuppressive diseases and its occurrence on canine distemper in vivo has not been published. In this study, the occurrence of apoptosis was determined in lymphoid tissues of thirteen naturally infected dogs and nine experimentally inoculated puppies. Healthy dogs were used as negative controls. Samples of lymph nodes, thymus, spleen and brain were collected for histopathological purposes. Sections, 5 microm thick, of retropharingeal lymph nodes were stained by HE, Shorr, Methyl Green-Pyronin and TUNEL reaction. Shorr stained sections were further evaluated by morphometry. Canine distemper virus nucleoprotein was detected by immunohistochemistry. Retropharingeal lymph nodes of naturally and experimentally infected dogs had more apoptotic cells per field than controls. In addition, DNA from thymus of infected dogs were more fragmented than controls. Therefore, apoptosis is increased in lymphoid depletion induced by canine distemper virus and consequently play a role in the immunosuppression seen in this disease. PMID- 12536302 TI - Complete nucleotide sequence of spring beauty latent virus, a bromovirus infectious to Arabidopsis thaliana. AB - Spring beauty latent virus (SBLV), a bromovirus, systemically and efficiently infected Arabidopsis thaliana, whereas the well-studied bromoviruses brome mosaic virus (BMV) and cowpea chlorotic mottle virus (CCMV) did not infect and poorly infected A. thaliana, respectively. We constructed biologically active cDNA clones of SBLV genomic RNAs and determined their complete nucleotide sequences. Interestingly, SBLV RNA3 contains both the box B motif in the intercistronic region, as does BMV, and the subgenomic promoter-like sequence in the 5' noncoding region, as does CCMV. Sequence comparisons of SBLV, BMV, CCMV, and broad bean mottle virus demonstrated that SBLV is closely related to BMV and CCMV. PMID- 12536303 TI - Identification of porcine alveolar macrophage glycoproteins involved in infection of porcine respiratory and reproductive syndrome virus. AB - The aim of this study was to identify the receptor(s) for PRRSV on porcine alveolar macrophages (PAMs) by producing monoclonal antibodies (MAbs) against these cells. Hybridoma supernatants were selected for their ability to block PRRSV infection. Four MAbs, 1-8D2, 9.4C7, 9.9F2, and 3-3H2 inhibited infection and recognised cell surface, PAM-specific antigens as shown by immunofluorescence and immunoperoxidase monolayer assay. These MAbs were then used to identify cellular proteins involved in PRRSV infection by radioimmunoprecipitation assays (RIPAs). MAbs 1-8D2 and 9.9F2 each recognised a 150 kDa-polypeptide doublet, while MAbs 9.4C7 and 3-3H2 both recognised a 220 kDa-polypeptide. Glycosidase treatment demonstrated all these polypeptides to be N-glycosylated. Thus, multiple glycoproteins appear to be involved in infection of PAMs by PRRSV. PMID- 12536304 TI - The complete nucleotide sequence of Pepper mottle virus-Florida RNA. AB - The Pepper mottle virus-Florida (PepMoV-FL) RNA genome was cloned and sequenced, and shown to consist of 9,717 nucleotides (nt) excluding the poly (A) tail. A single open reading frame was identified beginning at nucleotide position 169 encoding a polyprotein of 3068 amino acids. Phylogenetic sequence analysis revealed that of 44 full-length viral RNA genomes analyzed within the family Potyviridae, PepMoV-FL was most closely related to PepMoV-California (PepMoV-CA), Potato virus Y-H (PVY-H), PVY-N, PVY(o) and Potato virus V-DV42 (PVV-DV42). Using the PepMoV-FL sequence as a basis for comparison, the overall nucleotide sequence identity was highest between PepMoV-FL and PepMoV-CA at 93%, while the relationship was more distant with PVV-DV42 at 64% and for the PVY strains at 61%. A unique direct repeat sequence of 76 nucleotides was identified in the PepMoV-FL 3'-untranslated region (UTR), and this repeat sequence was confirmed not to occur in the PepMoV-CA sequence. Since the Florida isolate was among the first of the PepMoV isolates described, extensive biological and serological data on this isolate are available, and it has now been cloned and sequenced, we recommend that PepMoV-FL be recognized as the PepMoV type strain. PMID- 12536305 TI - An unusual large intergenic region in the S-RNA of a Bulgarian tomato spotted wilt virus isolate. AB - The complete S-RNA sequences of four Bulgarian and one German isolate of tomato spotted wilt tospovirus were determined. All isolates show a high conservation in their N proteins, while the NSs proteins and the intergenic regions (IGR) are more variable. The Bulgarian isolate 10HK96 has the largest S-RNA (3364 nucleotides) among tomato spotted wilt viruses reported so far. The enlargement is based on an insertion of 365 nts in the IGR that may have resulted from stuttering of the viral polymerase or non-homologous recombination. This insertion is present in the N protein gene subgenomic messenger, upstream of a proposed transcription termination signal. PMID- 12536308 TI - Molecular biology of laryngeal squamous cell carcinoma. AB - Some of the mechanisms involved in neoplastic transformation and progression of laryngeal squamous cell carcinoma (LSCC) are discussed. Although tumor suppressor inactivation of p53 and p16 is common in these tumors (about 50% each), oncogenic activation is less well characterized. Cyclin D1 and epidermal growth factor receptor amplification have been reported in one-third and one-quarter of LSCCs, respectively, both related to advanced stages, whereas c-myc could be amplified in 13% of cases although without associated overexpression. The role of ras in LSCC is, at most, exceptional, and the role of human papillomavirus infection in these neoplasms could have been largely overestimated. The AIS (amplified in squamous carcinoma) gene has been recently proposed as the main oncogenic target in head and neck squamous carcinomas and is a promising line of investigation. This, along with the link that exists between p53 and INK4 suppressor pathways through ARF and MDM-2, and the role of the universal cdk inhibitors (the Cip/Kip family) in these neoplasms deserve further investigation. Not forgotten are the mechanisms leading to cell immortalization and invasive capabilities acquisition, some of which are also briefly described. PMID- 12536309 TI - Parvovirus B19: a pathogen responsible for more than hematologic disorders. AB - The clinical and pathomorphological patterns of parvovirus B19 (PVB19)-associated diseases is the result of a balance between virus, host target cells and immune response. It is a characteristic feature of PVB19 that in patients with various other preexisting diseases, e.g., many hemolytic anemias, immune complex-mediated vasculitic disorders, and primary or secondary immunodeficiencies, the underlying diseases can be triggered, aggravated or complicated by severe organ manifestations. Identification of PVB19 by means of routine histology and immunohistology is only given in lytic infections occurring in transient aplastic anemia or nonimmune hydrops fetalis by the detection of viral inclusion bodies in erythroid precursor cells. In all other PVB19-associated diseases, molecular pathological methods must be applied. In this report, quantitative real-time polymerase chain reaction was used to determine the viral load in formalin-fixed and paraffin-embedded tissues derived from various organs. Using in situ hybridization it was demonstrated that endothelial cells of the microcirculatory periphery of the heart and hepatobiliar system in lytic infections are PVB19 specific target cells in children and adults. Because treatment of lytic PVB19 infection has been successfully applied, the pathologist should be alerted to include PVB19 into the diagnostic spectrum of viral disease, especially in immunocompromised patients. PMID- 12536310 TI - High-grade dysplasia and superficial adenocarcinoma in Barrett's esophagus: histological mapping and expression of p53, p21 and Bcl-2 oncoproteins. AB - In order to characterize the early morphological and molecular stages of the neoplastic progression of Barrett's mucosa, we performed the entire histological examination of ten specimens of resected Barrett's esophagus with high-grade dysplasia or superficial adenocarcinoma. The expression of p53, p21 and Bcl-2 proteins was assessed by immunohistochemistry. The surface of Barrett's mucosa ranged from 2.6 cm(2) to 31 cm(2). Dysplasia and adenocarcinoma always developed in specialized mucosa and often occupied small surfaces. High-grade dysplasia was multifocal in eight cases. There was no preferential site for neoplastic transformation into high-grade dysplasia or superficial adenocarcinoma in Barrett's mucosa. Three superficial adenocarcinomas and four high-grade dysplasias overexpressed p53 protein. p21 protein was focally expressed in nondysplastic mucosa and overexpressed in two superficial adenocarcinomas, one high-grade dysplasia and two low-grade dysplasias. In most cases, the expression of p21 and p53 proteins was unrelated. Bcl-2 protein was detected in only one area of low-grade dysplasia. In our study, high-grade dysplasia and superficial adenocarcinoma appeared as tiny lesions, often multifocal for high-grade dysplasia confirming the need for an extensive sampling of Barrett's mucosa in the endoscopic surveillance. p53 dysfunction plays a major role in the progression from dysplasia to carcinoma in Barrett's esophagus and appears unrelated to p21 and Bcl-2 expression. PMID- 12536311 TI - Acral myxoinflammatory fibroblastic sarcoma: a report of five cases and review of the literature. AB - Five cases of acral myxoinflammatory fibroblastic sarcoma that occurred in the distal extremities within the subcutaneous tissue are described. In one case, recurrence and metastases were recognized rather rapidly, only 3 months after the first excision. There have been no reports of early recurrence or metastases, especially the latter. The predominant type of constituent cells, cellularity of the neoplastic cells and density of inflammatory cells varied microscopically among cases. However, characteristic ganglion-like cells, Reed-Sternberg-like cells, round mononuclear cells and myxoid stroma, sometimes only seen focally, were found in all cases. Positive immunoreaction for vimentin was present in all cases. There was no correlation between positivity of MIB-1 or p53 for the primary tumor and presence of recurrence or metastases. In conclusion, we should be more cautious about the possibility of recurrence or metastases in earlier phases of acral myxoinflammatory fibroblastic sarcoma. Identification of the atypical bizarre fibroblastic component as the manifestation of the malignant nature of this lesion is vital to correct diagnosis, and it is important to attend to the myxoid and hyalinized zones, the inflammatory infiltrate, the presence of ganglion-like cells and acral location as features of acral myxoinflammatory fibroblastic sarcoma. PMID- 12536312 TI - Actin isoform pattern expression: a tool for the diagnosis and biological characterization of human rhabdomyosarcoma. AB - The diagnosis and characterization of rhabdomyosarcoma requires the use of combined histological and immunohistochemical criteria due to the variety of its histological patterns. The identification of actin isoform expression is accepted as a useful adjunct to the diagnosis and classification of soft tissue tumors. Using a new antibody specific for alpha-cardiac actin, obtained according to a recently described strategy for the production of polyclonal antibodies against actin isoforms [9], we have analyzed a series of 17 rhabdomyosarcomas, including all histological subtypes. In addition, we have evaluated the presence in these tumors of alpha-skeletal and alpha-smooth muscle actins. All specimens examined revealed a positive immunostaining for alpha-cardiac actin. Tumoral cells of eight cases also expressed alpha-smooth muscle actin and only three cases (all embryonal subtypes) were positive for alpha-skeletal actin. Our results indicate that immunohistochemical screening for alpha-cardiac actin expression is a useful tool for the diagnosis of rhabdomyosarcoma. They also suggest that the expression of alpha-skeletal actin is valuable in determining the subtype and possibly the state of differentiation of these tumors. PMID- 12536313 TI - Tissue remodeling following submassive hemorrhagic necrosis in rat livers induced by an intraperitoneal injection of dimethylnitrosamine. AB - We examined regeneration and fibrosis in the necrotic areas of hepatic stellate cells (HSCs). Acute hepatic injury was induced in rats by administration of an intraperitoneal injection of high-dose dimethylnitrosamine (50 mg/kg body weight). Liver samples were obtained from rats 6, 12, 24, 36 h and 2, 3, 5, 7, 10, and 14 days after the injection. They were examined by light and electron microscopy and by immunohistochemical methods. Hemorrhagic necrosis became most prominent 36 h after treatment and extended into zones 3 and 2. In the submassive necrotic areas the sinusoidal structure was destroyed. No HSCs positive for alpha smooth muscle actin or desmin were present. On day 5, when necrotic tissues were almost removed by infiltrating macrophages, HSCs strongly positive for alpha smooth muscle actin and desmin appeared along the surface of the preserved parenchyma and migrated into the necrotic areas along the residual reticulin fibers. By day 14 most of the necrotic areas were almost completely replaced by the regeneration of hepatocytes and central to central (C-C) bridging fibrosis. Our results indicate that following submassive complete necrosis, HSCs in the preserved liver parenchyma have roles in the formation of sinusoidal wall for remodeling in necrotic areas via their activation, proliferation, and migration into the necrotic areas. PMID- 12536314 TI - Diffuse mesenterial sclerosis: a characteristic feature of chronic small-bowel allograft rejection. AB - Chronic rejection is the major cause of late intestinal allograft dysfunction. The aim of this study was to analyze in detail the histopathological features of chronic rejection in the ACI-to-Lewis rat model of intestinal transplantation. Chronic rejection was achieved in orthotopic small-bowel allografts (ACI-Lewis) by limited immunosuppression with cyclosporin A (CyA). Isogeneic transplants (ACI ACI) as well as native bowels (ACI) with and without immunosuppression served as controls. Bowels were removed together with the mesenteries 90 days postoperatively and analyzed using sections stained with hematoxylin and eosin as well as Masson's trichrome. The slides were coded, randomized and analyzed by grading of histological abnormalities. The most striking alterations of the allografts were noticed in the mesenteries exhibiting an extensive infiltration by mononuclear cells accompanied by a progressive diffuse fibrosis with shrinking of the mesenteries. These changes were most pronounced in the perivascular areas of the mesenteric arteriae and venae rectae. Three of five allografts showed vasculitis with myointimal proliferation of the arteriae rectae. Focally, there was spill-over of the inflammatory cells onto the intestinal muscularis propria. The mucosa of the allografts showed mild blunting, lymphocytic infiltration of the crypt epithelium and increased crypt cell apoptoses. The submucosa was unaffected, and there were no detectable abnormalities of the enteric ganglion cells. The present data support the view that chronic rejection of intestinal allografts is characterized by a diffuse sclerosing mesenteritis which may significantly contribute to late graft dysfunction. The present model may be useful to study the pathomechanisms of this inflammatory fibrosing process. PMID- 12536315 TI - Increase of inactive intra-alveolar surfactant subtypes in lungs of asthmatic Brown Norway rats. AB - We tested the hypothesis whether allergic airway inflammation in ovalbumin sensitized and challenged Brown Norway rats is associated with intrinsic surfactant alteration and dysfunction. The determination of intra-alveolar surfactant subtypes and alveolar edema within their original microenvironment is only possible using an ultrastructural stereological approach. Therefore both lungs of control and asthmatic rats were fixed by vascular perfusion. The volume fractions of surfactant subtypes and the epithelial surface fraction covered with alveolar edema were determined by point and intersection counting. Furthermore, lung resistance was measured by means of whole-body plethysmography. The surface activity of surfactant from bronchoalveolar lavage was determined as minimum surface tension at minimal bubble size with a pulsating bubble surfactometer. Compared with controls, in asthmatics (1) the fraction of inactive unilamellar forms was significantly increased from 56% to 66%, (2) the fraction of alveolar epithelium covered with alveolar edema visible by light microscopy was significantly increased from 0.7% to 5.0%, (3) the fraction of alveolar epithelium covered with fluid seen by electron microscopy expanded significantly from 5% to 21%, (4) lung resistance was significantly elevated from 14% to 86% and (5) surface tension was enhanced from 6 mN/m to 12 mN/m. Thus, the inflammatory process after allergen challenge of sensitized Brown Norway rats causes intra-alveolar surfactant alterations. These surfactant alterations might contribute to small airway dysfunction. PMID- 12536317 TI - Minimally invasive (encapsulated) follicular carcinoma of the thyroid gland is the low-risk counterpart of widely invasive follicular carcinoma but not of insular carcinoma. AB - Minimally invasive (encapsulated) follicular carcinoma (MIFC), widely invasive follicular carcinoma (WIFC), and insular carcinoma (IC) are three distinct types of thyroid carcinomas composed of follicular cells, IC representing the poorly differentiated counterpart. Our aim was to test whether there is a time-dependent relationship among these three entities, in particular whether MIFC is the low risk form of the entire group. We used overall survival analysis and multiple correspondence analysis (MCA), the latter exploring the clinicopathological characteristics at onset (sex, age, TNM/pTNM classification). The study included 18 MIFCs, 29 WIFCs, and 27 ICs. On the basis of both overall survival and MCA, WIFC emerged as the high-risk counterpart of MIFC, occurring in older age, with a larger size, a widespread growth into the thyroid parenchyma, and presenting with distant metastases. In both types of follicular carcinoma distant metastases involved only lung and bone, and there was no tendency for local spread beyond the thyroid capsule or nodal metastases. By contrast, IC confirmed to be a distinct high-risk lesion, showing both follicular and papillary-related biological behavior characteristics. PMID- 12536316 TI - Expression of carbonic anhydrase-related protein CA-RP VIII in non-small cell lung cancer. AB - Carbonic anhydrase-related protein (CA-RP VIII) lacks a Zn-binding motif which is essential for carbonic anhydrase activity. Therefore, CA-RP VIII is believed to have a no catalytic activity and a new biological property. In the present study, CA-RP VIII expression in non-tumorous lung and non-small cell lung carcinomas was investigated. Little or no expression of CA-RP VIII was observed in human lungs by Northern-blot analysis. Reverse-transcription polymerase chain reaction analysis demonstrated CA-RP VIII mRNA in developing human lungs and, to a lesser extent, in normal lungs. Subsequent immunohistochemical staining revealed that CA RP VIII was expressed in the pulmonary epithelium in developing lungs; however, CA-RP VIII expression was restricted in bronchial ciliated cells in adult lungs. Neither bronchial gland nor squamous metaplasia of interstitial pneumonia expressed CA-RP VIII. In contrast, CA-RP VIII was strongly expressed in almost all archival lung cancer specimens, which included 24 squamous cell carcinomas, 25 adenocarcinomas, and 6 adenosquamous cell carcinomas. Cancer cells at the front of tumor progression expressed CA-RP VIII in particular abundance. The present findings suggest that CA-RP VIII may play a role in non-small lung cell carcinomas. PMID- 12536318 TI - A gastric carcinosarcoma with neuroendocrine cell differentiation and undifferentiated spindle-shaped sarcoma component possibly progressing from the conventional tubular adenocarcinoma; an immunohistochemical and ultrastructural study. AB - A 56-year-old Japanese man underwent total gastrectomy with lymph-node dissection for a gastric tumor. In the oral rim of the excised large ulcerated tumor, a small nodular mass demonstrated a unique histology of a carcinosarcoma composed of more than three distinctive components. A major part of the large ulcerated tumor tissue consisted of conventional tubular adenocarcinoma, with the coincidental focal distribution of solid cell nests of poorly differentiated or neuroendocrine cell (small cell) carcinoma. Sarcoma cells were another of the constituents of the small nodular tumor with atypical spindle-shaped cells. All the excised lymph-node metastasis demonstrated the histology of tubular adenocarcinoma. In the sarcoma cells, many of the lineage-specific immunohistochemical markers were negative. Only vimentin was positive. Electron microscopy revealed that the neuroendocrine cell carcinoma contained cells with abundant small aggregations of neurosecretory-type granules in their cellular cytoplasm. Primitive cellular junctions and incomplete basal lamina were seen in the sarcoma cells. No transitional or hybrid-type cells were seen between carcinoma and sarcoma cells. The interfaces between the cancer cell nests and the proliferation of sarcoma cells are clearly discerned. It was speculated that the unique histology of carcinosarcoma might have progressed from the conventional type of pre-existing advanced tubular adenocarcinoma. PMID- 12536319 TI - Malakoplakia of the pancreas with diffuse lymph-node involvement. AB - We report a case of malakoplakia involving the pancreas in a 74-year-old man with associated regional lymphoadenopathy. Histological examination of both pancreas and lymph nodes revealed a diffuse histiocytic infiltrate containing numerous Michaelis-Gutmann bodies. Electron microscopy supported the diagnosis of malakoplakia and showed bacterial-like structures. Differential diagnosis includes myofibroblastic inflammatory tumor and histiocytic neoplasms. Lymph-node involvement during malakoplakia is extremely rare and it has never been documented microscopically. Lymphohematogenous spread of bacteria may be the cause of the nodal involvement, which, however, does not appear to influence the clinical course of the disease. PMID- 12536320 TI - The ancestral role of Brachyury: expression of NemBra1 in the basal cnidarian Nematostella vectensis (Anthozoa). AB - The T-Box transcription factor Brachyury plays important roles in the development of all bilateral animals examined so far. In order to understand the ancestral function of Brachyury we cloned NemBra1, a Brachyury homolog from the anthozoan sea anemone Nematostella vectensis. Anthozoa are considered the basal group among the Cnidaria. First NemBra1 expression could be detected at the blastula/gastrula transition and gene activity persists until adulthood of the animals. In situ hybridization shows that NemBra1 expression in gastrulae and early planula larvae is restricted to a circle around the blastopore. When the larvae begin to metamorphose into primary polyps, the expression zone extends into the developing mesenteries. In adult polyps Brachyury expression persists in the mesenteries, but is excluded from the septal filament and the differentiated retractor muscles, which also develop from the mesenteries. We conclude that the ancestral function of Brachyury was in specifying the blastopore and its endodermal derivatives. PMID- 12536321 TI - Localization of ecdysone receptor protein during colour pattern formation in wings of the butterfly Precis coenia (Lepidoptera: Nymphalidae) and co-expression with Distal-less protein. AB - Butterfly wing colour patterns are determined during late larval and early pupal development, a part of metamorphosis controlled by ecdysteroid hormones via their nuclear hormone receptors. We have sequenced a fragment of the common regions of the ecdysone receptor (EcR) from the butterflies Precis coenia and Bicyclus anynanaand found high identities (83.5% to 100%) to EcR from a moth, Manduca sexta. In P. coenia, we sequenced a putative EcR-B1 isoform with 80.4% identity with the A/B-region of the M. sexta-EcR-B1. Consequently, we used antibodies generated against MsEcR-B1 to localise EcR protein during wing development of P. coenia. Nuclear staining of EcR was observed in different cell types during the course of colour pattern formation. Major observations are as follows: EcR is expressed in cell nuclei corresponding to wing lacunae and prospective veins. EcR is expressed early in pupal wing development in "focal" cells which are thought to release determining signals in a process leading to eyespot formation. Scale forming cells differentiate first and show EcR signal in the eyespot foci and most of the wing sheet, but not in areas corresponding to prospective eyespots. In the eyespots, 20-24 h after pupation, EcR expression seems to play a role in formation of scale rows preceding a later expression (28 h) in scale-forming cells. The results demonstrate that EcR is locally expressed in correlation to all major events of wing development and colour pattern formation. In particular, EcR expression patterns in prospective eyespots show that these are special pattern elements which are specified in concert with other factors of colour pattern formation such as the transcription factor Distal-less. In eyespot foci, Distal-less is expressed simultaneously with EcR, but clearly precedes EcR expression in eyespot scale-forming cells. This indicates a potential interaction between "short-range" signalling systems and "long-range" hormonal systems. PMID- 12536322 TI - Transcriptional pattern of a novel gene, expressed specifically after the point of-no-return during sexualization, in planaria. AB - We have investigated sexualization of asexual worms in the planarian Dugesia ryukyuensis. During sexualization there is a point from which an animal cannot return to the asexual state (point-of-no-return). To isolate the genes related to the point-of-no-return, we performed differential screening and isolated one novel gene that was expressed specifically in yolk glands of the worms after the point-of-no-return and named it Dryg. It encoded 655 amino acids with a predicted molecular mass of 79 kDa. We performed a series of experiments using Dryg as a molecular marker in the yolk gland. At first, we monitored how the yolk gland was formed during sexualization. The expression in sexualizing worms at stage 3 is limited to a single type of cell that has characteristics of neoblasts, the totipotent somatic cells; however, the expression is observed in the yolk gland in sexualized worms. Furthermore, we monitored yolk glands for expression during regeneration. The original yolk glands seem to disappear after ablation, then new yolk glands appeared along the ventral nerve cords. Because this expression pattern looks like that of sexualizing worms at stage 3, we speculate that yolk gland cells may differentiate from neoblasts during regeneration as observed during sexualization. PMID- 12536323 TI - Phenylthiourea disrupts thyroid function in developing zebrafish. AB - Thyroid hormone (T4) can be detected in thyroid follicles in wild-type zebrafish larvae from 3 days of development, when the thyroid has differentiated. In contrast, embryos or larvae treated with goitrogens (substances such as methimazole, potassium percholorate, and 6-n-propyl-2-thiouracil) are devoid of thyroid hormone immunoreactivity. Phenythiourea (PTurea; also commonly known as PTU) is widely used in zebrafish research to suppress pigmentation in developing embryos/fry. PTurea contains a thiocarbamide group that is responsible for goitrogenic activity in methimazole and 6-n-propyl-2-thiouracil. In the present study, we show that commonly used doses of 0.003% PTurea abolish T4 immunoreactivity of the thyroid follicles of zebrafish larvae. As development of the thyroid gland is not affected, these data suggest that PTurea blocks thyroid hormone production. Like other goitrogens, PTurea causes delayed hatching, retardation and malformation of embryos or larvae with increasing doses. At doses of 0.003% PTurea, however, toxic side effects seem to be at a minimum, and the maternal contribution of the hormone might compensate for compromised thyroid function during the first days of development. PMID- 12536324 TI - Vasa expression and germ-cell specification in the spider mite Tetranychus urticae. AB - The specification of germ cells is an important process during the development of all animals. Expression of an evolutionarily conserved gene such as vasa can be used as a marker for germ cell fate. We have isolated a vasa-related gene from the two-spotted spider mite (Tetranychus urticae) and used it to examine the segregation of germ cells in this animal. In spider mites, vasa expression first appears in a group of cells that do not join the initial blastoderm surface. Instead, these cells remain in the interior of the blastoderm and then migrate to posterior regions of the embryo, where they form a cluster that appears in regions of the embryo consistent with the gonads. The expression pattern of this spider mite vasa homologue implies a novel process acts to specify germ cells in this species and that the specification of germ cells is an evolutionarily labile process. PMID- 12536325 TI - Developmental expression of the Xenopus laevis Tbx20 orthologue. AB - We have isolated the Xenopus orthologue of the T-box gene, Tbx20, and characterized its developmental expression profile. We show that Tbx20 is one of the earliest markers of heart tissue in Xenopus, and is expressed throughout all cardiac tissue during later stages of development. In addition, we also observe expression in the cement gland, the jugular vein, the lung bud, the cloacal aperture, rhombomeres 2, 4, 6 and 8, and in a subset of motor neurons. PMID- 12536326 TI - The Xenopus arx gene is expressed in the developing rostral forebrain. AB - The human aristaless-related homeobox ( ARX) gene is mutated in several patients with X-linked mental retardation and/or other neurologic pathologies. We report the isolation and expression pattern of a Xenopus arx gene. Similar to other vertebrate arx genes, Xenopus arx is expressed in the developing telencephalon, diencephalon, and floor plate. PMID- 12536327 TI - Endoscopic versus operative treatment in high-risk ulcer bleeding patients - results of a randomised study. AB - BACKGROUND AND AIMS: The treatment of peptic ulcer bleeding is still a matter of controversy in high-risk patients with arterial bleeding or a visible vessel. Patients with recurrent bleeding during hospital stay carry an increased risk of death. Therapeutic concepts using early elective operation compete with solitary endoscopic treatment. Numerous prospective studies have contrasted comparable improvements for the different therapeutic regimens but there is still a lack of a randomised comparison. PATIENTS AND METHODS: We initiated a multicentre randomised clinical trial comparing endoscopic fibrin glue injection with early elective operation in peptic ulcer patients with arterial bleeding or a visible vessel > or =2 mm. After initial endoscopic control of bleeding, patients were randomised to repeated fibrin glue injection or early elective operation. Outcome criteria were recurrent bleeding and death. The study was terminated after a planned interim analysis. RESULTS: Due to strict inclusion and exclusion criteria 61 patients were randomised and 55 patients could be included in the per-protocol analysis, 23 in the early elective operation group and 32 in the patient group with endoscopic therapy. The type of surgery in the early elective operation group was usually gastric resection (79%). Recurrent bleeding occurred in 50% of the endoscopically treated patient group, and in the operative group in one patient (relative risk: 11.5; 95% CI: 1.6 to 80.7). There were no statistically significant differences between the two treatment groups with respect to mortality (relative risk: 0.7, 95% CI: 0.1 to 4.8). CONCLUSION: Early elective surgery is an effective procedure in bleeding peptic ulcer patients at high risk for re-bleeding. Fibrin glue injection carries a risk for re-bleeding, however, the majority of these re-bleeding episodes can be controlled by re-endoscopic treatment, but a subgroup will need an emergency operation with a fatal outcome in individual patients. PMID- 12536328 TI - Cystic duct carcinoma: a proposal for a new "working definition". AB - BACKGROUND: Farrar's criteria for cystic duct carcinoma (histopathological diagnosis of a carcinoma strictly limited to the cystic duct) have practical limitations. We propose new "working definition": a gallbladder tumor, the center of which is located in the cystic duct. PATIENTS AND METHODS: Between 1980 and 2000 we diagnosed cystic duct carcinoma in 31 patients, 28 of whom (90%) had increased serum bilirubin concentrations. Extrahepatic bile duct resection and cholecystectomy were performed in 10 patients; in others, extended right hepatectomy (16), right hepatectomy (3), and liver bed resection (2) were necessary as well for a potentially curative resection. Additional procedures were portal vein resection (10) and pancreatoduodenectomy (7). RESULTS: All tumors were adenocarcinomas. Depth of invasion was T2 in 3 patients, T3 in 12, and T4 in 16. Thirteen patients (42%) had lymph node metastasis. Curative resection was performed in 24 patients (77%). Hospital mortality was 5 of 31 (16%). Actuarial 5-year survival rate excluding hospital deaths was 22%. CONCLUSIONS: The proposed "working definition" avoids the problems associated with Farrar's criteria and describes a distinct patient group with an approximately equal proportion of men and women, advanced T stage, but a lower than expected frequency of lymph node metastasis. It establishes a basis for standard reporting of results. PMID- 12536330 TI - Operative treatment of renal autonomous hyperparathyroidism: cause of persistent or recurrent disease in 304 patients. AB - BACKGROUND: Subtotal parathyroidectomy (SPTX) and total parathyroidectomy with autotransplantation (TPTX and AT) are standard procedures in the treatment of renal autonomous hyperparathyroidism. In contrast to primary hyperparathyroidism, the persistence/recurrence rate is reported of up to 12.0%. PATIENTS AND METHODS: Between 1986 and 2000 we operated on 304 patients with renal autonomous hyperparathyroidism including 14 patients who were admitted after a primary operation in an outside hospital. Mean observation period was 51.4+/-38.9 months. RESULTS: The overall persistence/recurrence rate in our patients was 9.0% (26/290). After SPTX, excluding patients with an incomplete operation, it was 3.7%, and after TPTX and AT it was 6.0%. Reasons for developing recurrent or persistent disease in these patients were removal of less than 3.5 glands ( n=12), hyperplastic autograft ( n=5), and supernumerary gland ( n=4). After the first reoperation 7 patients (26.9%) had persistent or recurrent disease. CONCLUSIONS: An incomplete primary operation caused by missed cervical glands was the major reason for persistent ( n=8) or recurrent ( n=4) disease after different operative strategies in renal autonomous hyperparathyroidism. PMID- 12536329 TI - Liver abscess after non-operative management of blunt liver injury. AB - BACKGROUND: The non-operative management of blunt liver trauma can be applied in almost 80% of patients with this type of injury, with the advantages of the need for fewer blood transfusions, less intra-abdominal sepsis, and a better survival rate, than with the operative approach. However, liver abscess, as a known complication of the non-operative management of blunt liver trauma, is discussed infrequently. Therefore, we herein review our experience and describe this complication in detail. MATERIALS AND METHODS: From 1995 to 2001, 674 patients were admitted to our hospital due to blunt hepatic trauma. Among these patients, 279 underwent laparotomy and the remaining 395 patients were treated non operatively. Twenty-two patients were identified as having liver abscess, with 16 of them belonging to the operative group, and six to the non-operative group. A retrospective review of these six patients and their characteristics, as well as pathogenesis, diagnosis, and the management of the liver abscesses, was conducted. RESULTS: These six patients were all male, with a median age of 19.5 years (range 3-24). The median injury severity score was 16.5 (range 9-25); three patients sustained grade-3 hepatic injury, and the other three were grade 4. The main diagnostic tool was abdominal computed tomography, and the abscesses took a median of 6 days (range 1-12) to form and be diagnosed. The abscesses were usually caused by infection from mixed organisms, and an abscess resulting from Clostridium infection developed within 1 day after injury. These abscesses were treated with antibiotics and drainage, and the median length of hospital stay was 26 days (range 8-44), without mortality or long-term morbidity. CONCLUSION: Liver abscess as a complication of the non-operative management of blunt hepatic trauma is a rare entity, with an incidence of 1.5% (6/395). It is usually seen in severe liver injury (grade 3 and above), but all our patients were all treated successfully, with no mortality. However, prolonged hospitalization may be required in this patient group. PMID- 12536331 TI - Reimplantation of extracorporeal irradiated bone segments in musculoskeletal tumor surgery: clinical experience in eight patients and review of the literature. AB - BACKGROUND: Two methods are available for extracorporeally devitalizing resected tumor-bearing bone specimens, the simpler of which, autoclaving, has substantial disadvantages. We present our experience with the alternative reconstruction technique of reimplantating extracorporeally irradiated specimens (IEIR) PATIENTS AND METHODS: Eight patients who had primary malignant bone tumors of the long bones were managed with wide en bloc resection and IEIR. The segments were fixed by either plate osteosynthesis, knee arthrodesis rod, or intramedullary nailing. All seven patients with high-grade tumors received chemotherapy. RESULTS: After a median follow-up of 66 months (40-76) five of eight patients were continuously free of disease. One man with a chondroblastic osteosarcoma developed pulmonary metastases which were resected. One woman with teleangiectatic osteosarcoma developed a local recurrence in the soft tissue without contact to the irradiated reimplant. At the latest follow-up, 58 months after resection of the pulmonary metastases and 28 months after resection of the locally recurrent tumor, there was no evidence of disease in either patient. Another woman 67 months after IEIR for an osteosarcoma of the distal femur developed a subcutaneous metastasis of the scalp and the thoracic wall and an ossifying pulmonary metastasis. At the time of writing she is receiving chemotherapy. After a median duration of 4.3 months (2.3-25.2) all graft-host junctions had healed. The functional result was good in four patients and excellent in the other four. CONCLUSIONS: Limb salvage using reimplantation of extracorporeally irradiated tumor-bearing bone segments for reconstruction offers an excellent reconstruction method in appropriately selected patients compared with other options of management. PMID- 12536332 TI - Extent of oncological resection in laparoscopic vs. open colorectal surgery: meta analysis. AB - BACKGROUND: Laparoscopy is not yet wide accepted in cancer colorectal surgery. This study investigated whether the extent of anatomical resection in laparoscopic colorectal surgery is equivalent to that in the open colorectal surgery. MATERIALS AND METHODS: We examined reports on the results of laparoscopic or open colorectal procedures published between 1990 and 1999 and selected 35 with data on lymph node count and distal margin clearance (total of 3935 patients). There were 16 comparative studies, 6 open series, and 13 laparoscopic series of patients. Several meta-analytic models were used to evaluate the difference between open and laparoscopic surgery, including multiple outcome random-effects models that account for correlation between multiple outcomes. RESULTS: More lymph nodes were extracted laparoscopically (0.3-2.14 lymph nodes more); however, only for the fixed-effects single-outcome model this difference was statistically significant. The average distal margin clearance was 4.6 cm with the laparoscopic approach and 5.3 with the open approach Accounting for random variation among the studies, the estimate of this difference obtained from meta-analysis was 0.64-1.19 cm, favoring the open approach; this difference was statistically significant. The estimated within-study correlation between lymph node count and distal margin clearance was 0.26 with the open and 0.19 with the laparoscopic procedure. The estimated within-study correlation between outcomes of laparoscopic and open procedures was 0.86 for the lymph node count and 0.96 for the distal margin clearance. CONCLUSIONS: This meta-analysis shows that laparoscopic approach is as adequate as the conventional approach. Estimates of difference between the two therapeutic options obtained from random-effects models were more precise than those from fixed-effects models. PMID- 12536333 TI - Plasma vitamin E and A changes during cardiopulmonary bypass and in the postoperative course. AB - BACKGROUND: Cardiopulmonary bypass induces a generalized inflammatory reaction accompanied by free radical generation. Depletion of antioxidants could result and is reported for vitamin E and C. We investigated the effect of cardiopulmonary bypass on plasma concentrations of alpha-tocopherol, retinol, and biochemical variables (e.g., triacylglycerol, cholesterol, and C-reactive protein). PATIENTS AND METHODS: Plasma levels of all parameters were investigated by serial sampling in ten men undergoing elective coronary artery bypass grafting. Samples were taken before, during, and up to 48 h after bypass to obtain time profiles of the laboratory indices. RESULTS: alpha-Tocopherol and retinol decreased during cardiopulmonary bypass when not adjusted for confounders. After adjustment for hemodilution and lipids, no significant change was noted during bypass. However, a reduction in retinol was observed 48 h postoperatively. CONCLUSIONS: These data indicate that vitamin E and A analysis to ascertain links to their consumption via the production of free radicals under conditions of cardiopulmonary bypass may be inappropriate. Specifically, during bypass a reduction in systemic vitamin E and A seems to be a response to changes in blood volume and liver function. PMID- 12536334 TI - Splenic tissue autotransplantation in rabbits: no restoration of host defense. AB - BACKGROUND: The loss of spleen may increase the incidence of overwhelming sepsis. To prevent this, splenic autotransplantation has been performed in humans and experimental animals. However, there is still controversy about the effectiveness of regenerated splenic tissue in preventing infection. This study explored the effectiveness of splenic tissue autotransplantation in restoring host defense. MATERIALS AND METHODS: Rabbits were divided into three groups: splenic autotransplantation, sham operation, and total splenectomy. Histomorphology, T lymphocyte count, serum lysozyme levels, hemolysin titers, and pneumococcal clearance were observed as read-out parameters over 24 weeks. RESULTS: Histological study showed that the white pulp was poorly developed and central arterioles were missing in the regenerated splenic tissue of the autotransplanted rabbits. The weight of regenerated spleens recovered 6 months later in the splenic autotransplantation group was 11% of that in the sham operation group and was significantly less than the weight at implantation. There was no significant difference in the number of T lymphocytes or level of serum lysozyme between the three groups. A poor antibody response by the rabbits in the splenic autotransplantation and total splenectomy groups was noted after the primary intravenous administration of sheep red blood cells compared to those of sham operation group. After the challenge with type 3 pneumococci intravenously, pneumococcal clearance from the bloodstream in the splenic autotransplantation group did not differ significantly from that in the total splenectomy group, but was markedly delayed compared with that in the sham operation group. CONCLUSIONS: The low quantity and poor quality of the regenerated splenic tissue contribute to the inferior immunoprotective ability of animals autotransplanted with one-third of the original spleen. This suggests that the regenerated spleen cannot compensate for the immunological function of the original one, especially host resistance to infection. PMID- 12536335 TI - Scorecard endoscopy: a pilot study to assess basic skills in trainees for upper gastrointestinal endoscopy. AB - BACKGROUND: The development of training models and structured training courses for endoscopic techniques provides practical experience. To assess individual performance and progress in this training we developed and tested a scorecard system. METHODS: Three test groups were compared: group 1, ten physicians without previous endoscopic experience; group 2, ten students, without endoscopic experience; group 3, a control group of experienced endoscopists. Groups 1 and 2 underwent 1 week of training with a theoretical introduction and practical demonstrations. They were assessed by the scorecard daily by an experienced tutor. The individual scores and learning curves of the two beginner groups were compared with those of the expert group using a biosimulation model was used. RESULTS: Each participant improved significantly during the 1-week course. Mean scores on the first day in groups 1-3 were, respectively, 26.7+/-10.7, 33.4+/ 5.3, and 72.0+/-5.8, and on day 6 they were 62.2+/-6.6, 63.4+/-7.6, and 86.6+/ 4.3. The difference between group 3 and the other two groups was significant but not that between groups 1 and 2. CONCLUSIONS: Training in endoscopy can be assessed using our training model and our scorecard protocol, which distinguishes between various levels of experience. In physicians beginning in the field of gastrointestinal endoscopy this approach could help to reduce risks to patients, shorten learning curves, and exclude unskilled individuals from further fruitless interventions. PMID- 12536336 TI - "Operative treatment of renal autonomous hyperparathyroidism: causes for persistent or recurrent disease in 304 patients" (comments on C. Dotzenrath et al.). PMID- 12536338 TI - Coagulation index to distinguish severe preeclampsia from normal pregnancy. AB - The purpose of this study was to establish a new coagulation index to distinguish severe preeclampsia from normal pregnancy using optimal coagulation parameters. The difference between platelet counts in early gestation and before delivery (DeltaPlt = [platelet counts before delivery] - [platelet counts in early gestation]) was calculated as an index of changes in platelet counts. Antithrombin (AT) activity, thrombin-antithrombin (TAT) complex, fibrin degradation products (FDP) D-dimer, and DeltaPlt were investigated in 72 cases of normal pregnant women in the third trimester of pregnancy and 56 cases of severe preeclampsia. The new coagulation index was calculated using multivariate logistic regression analysis. As a result, in a case using four parameters, the following formula was obtained: Y = (-0.019 x AT activity) + (0.067 x TAT) + (0.067 x D-dimer) + (-0.064 x DeltaPlt) + 0.706. According to this formula, coagulation indices in normal pregnant women and in patients with severe preeclampsia were -0.77 +/- 0.51 and 1.41 +/- 1.56, respectively (p < 0.0001). Among patients with severe preeclampsia, coagulation and fibrinolysis disorders before delivery were typical in patients terminated by cesarean section (coagulation index = 1.62 +/- 1.66) compared with those with successful vaginal delivery (coagulation index = 0.52 +/- 0.34) (p < 0.05). These facts suggest that an excessive hypercoagulable state is associated with the termination of pregnancy resulting from the aggravation of preeclampsia. From the viewpoint of coagulation and fibrinolysis disorders, the predictive value in order to decide the optimal time for the termination of pregnancy could be recommended when the coagulation index exceeded 1.20. PMID- 12536339 TI - The M235T variant of the angiotensinogen gene and the body mass index are useful markers for prevention of hypertension in pregnancy: a tree-based analysis of gene-environment interaction. AB - We sought to perform a tree-based analysis of lifestyle risk factors for hypertension in pregnancy (HP) with univariate and multivariate analyses. Seventy eight HP patients and 199 normal controls were recruited from primiparous women 20 to 34 years of age. Data from angiotensinogen (AGT) genotyping and data from a self-administered questionnaire about lifestyle were subjected to univariate and multivariate analyses. By dividing the subjects into two subgroups--those who possessed "the TT genotype of AGT" and "body mass index (BMI) < 24" and those who did not--we were able to examine the acquired risk factors for HP during pregnancy in these two groups. Multivariate analysis selected "mentally stressful condition" and "no antenatal training during pregnancy" in the former group, and "poorly balanced diet" in the latter group. Determination of factors obvious before pregnancy, such as genotype or prepregnancy BMI, may be useful for devising effective individualized strategies for preventing HP. PMID- 12536340 TI - An association between maternal smoking and preeclampsia in Japanese women. AB - In order to determine whether maternal smoking before or during pregnancy, or both, is associated with a reduced risk for preeclampsia in Japanese subjects, we conducted a case-control study that took other risk factors for preeclampsia into consideration. Seventy-one preeclampsia patients were matched with 142 controls for parity and age. Information from a self-administered questionnaire and clinical data such as maternal age, parity, family history of hypertension, prepregnancy body mass index (BMI), and pregnancy outcomes were analyzed. No significant difference was found between the groups in smoking rates before and during pregnancy (38.0 and 18.3% in preeclampsia patients and 38.0 and 16.9% in controls, respectively). However, classification of the subjects according to the presence of "prepregnancy high body mass (BMI > or = 24)" revealed a significant association between maternal smoking before pregnancy and preeclampsia in women with a prepregnancy high body mass (a smoking rate of 47.6% in patients with preeclampsia and 7.1% in controls, p < 0.05). This result suggests that there is a clear racial difference in the manifestation of preeclampsia with respect to the effect of smoking and that early intervention, particularly before pregnancy, to get obese women to stop smoking may be effective for preventing preeclampsia. PMID- 12536341 TI - Anticholinergics induce eclamptic seizures. AB - Severe preeclampsia is a pathophysiological disorder specific to pregnancy and characterized by vasoconstriction and hypercoagulability. Eclampsia (convulsion associated with preeclampsia) and hemolysis, elevated liver enzymes, and low platelet count associated with preeclampsia (HELLP syndrome) are serious complications in patients with severe preeclampsia. They are thought to be characterized by generalized vasoconstriction and reduction in blood flow to various organs that may be explained by increased sensitivity of the vascular smooth muscles and increased vasopressors. Liver involvement in eclampsia and preeclampsia is referred to as HELLP syndrome, and epigastric and right upper quadrant pain is often a symptom of severe preeclampsia and may be indicative of imminent convulsions. In addition, marked dilatation of the stomach and the colon is often demonstrated in these patients. These phenomena result from the hyperactivity of the sympathetic nervous system but are not caused by the hyperactivity of the parasympathetic nervous system. The authors experienced two cases of eclamptic seizures after the administration of an anticholinergic (scopolamine butylbromide) in patients with severe preeclampsia complicated by HELLP syndrome. Anticholinergics, blocking agents of the parasympathetic nervous system, can enhance the hyperactivity of the sympathetic nervous system; therefore, vasospasms of the vessels may be easily aggravated, and eclamptic seizures may be induced in patients with severe preeclampsia, especially in those complicated by HELLP syndrome. The administration of anticholinergics should be avoided in patients with severe preeclampsia, especially when there is epigastralgia. PMID- 12536342 TI - Gestational thrombocytopenia and pregnancy-induced antithrombin deficiency: progenitors to the development of the HELLP syndrome and acute fatty liver of pregnancy. AB - The syndrome of hemolysis, elevated liver enzymes, and low platelet count (HELLP syndrome) and of acute fatty liver of pregnancy (AFLP) do not have an abrupt onset. Thrombocytopenia or reduced antithrombin activity, or both, seen at presentation do not result from these complications. There are a small number of pregnant women who exhibit a gradual antenatal decline in platelet counts or antithrombin activity, or both, irrespective of the presence or absence of preeclampsia. Those who develop a profound decrease in either platelet counts or antithrombin activity are at an increased risk for developing perinatal aspartate aminotransferase (AST) elevation. Thrombocytopenia or reduced antithrombin activity, or both, precede the onset of these diseases. Therefore, monitoring of platelet counts and antithrombin activity during pregnancy is clinically useful for identifying women at an increased risk of the HELLP syndrome and AFLP. Because women with twin pregnancies are likely to exhibit a decrease in platelet counts or antithrombin activity, or both, compared with women with singleton pregnancies, HELLP syndrome and AFLP are more likely to occur in women with twin pregnancies. PMID- 12536343 TI - Perioperative changes in plasma antithrombin activity and platelet counts in patients undergoing gynecologic surgery. AB - The relationships among perioperative changes in plasma antithrombin (AT) activity, platelet counts, and clinical risk factors for postoperative thromboembolism has not been studied extensively in gynecologic patients. AT activity and platelet counts were determined preoperatively and on postoperative days 1 and 7 in 74 patients who underwent gynecologic surgery. The relationships between each clinical risk factor for postoperative thromboembolism and the decrease in AT activity and platelet count were analyzed using a univariate logistic regression model. No clinically significant thromboembolism occurred in any patient. AT activity and platelet counts decreased significantly on postoperative day 1 and exceeded preoperative levels on day 7. Postoperative patients with malignancy were more likely to exhibit decreased AT activity. AT activity decreased significantly with increases in operating time and blood loss. Known risk factors for postoperative thromboembolism did not appear to be associated with postoperative decreases in platelet counts. Transient hyperconsumption of AT and platelets occurs immediately after gynecologic surgery. Surgery-induced AT deficiency may contribute to postoperative thromboembolism in patients with long operating times and significant blood loss. PMID- 12536344 TI - A case of pregnancy with a history of paradoxical brain embolism. AB - Brain embolisms in younger persons are rare but are often caused by a paradoxical embolism, the embolic entry of a venous thrombus into the systemic circulation through a right-to-left shunt. A 27-year-old pregnant woman presented with hemiplegia that had been treated with an antiplatelet agent since the occurrence of a paradoxical brain embolism via the pulmonary arteriovenous fistula. A tendency of hypercoagulation is generally observed during pregnancy, so a patient with this condition has a strong risk factor for venous thromboembolism during pregnancy and even more so for arterial thromboembolism under the intense strain of labor, which is much stronger than that of the Valsalva maneuver. This case had been controlled well with an antiplatelet agent and an anticoagulant while the levels of coagulation and fibrinolytic factors were monitored and was followed by a successful pregnancy outcome. PMID- 12536345 TI - Influence of low- and high-molecular-weight plasminogen activators on the onset of labor and on the hemostatic system. AB - Measurement of activities of urinary (u-PA) and tissue plasminogen activator (t PA) and analysis of euglobulin lysis times (ELTs) and prekallikrein were performed simultaneously using the plasma of normal pregnant women and nonpregnant women. The activities of plasminogen activators were determined by the method of electrophoretic zymography. The activity of low-molecular-weight (LMW) u-PA was significantly decreased at the onset of labor (p = 0.05) compared with that during the latter half of pregnancy. The same tendency was observed in the analysis of prekallikrein. On the contrary, no significance was shown in activities of high-molecular-weight (HMW) u-PA or t-PA. It is suggested that the action of kallikrein on low-molecular-weight u-PA causes it to change into plasmin at the onset of labor. PMID- 12536346 TI - Improvement of vitamin K status of breastfeeding infants with maternal supplement of vitamin K2 (MK40). AB - The present study is aimed at evaluating the efficacy of maternal vitamin K2 supplementation on the vitamin K status of newborn infants using the measurement of des-gamma-carboxyprothrombin (PIVKA-II [protein induced by vitamin K absence]) and the hepaplastin test (HPT). PIVKA-II and HPT were measured at the 1st month of age in two groups: 31 infants with maternal vitamin K supplementation (15 mg/d Menatetrenone since the 14th day after parturition) (group 1) and 46 without maternal supplementation (group 2). All infants received vitamin K2 syrup twice within the 1st week of life. The PIVKA-II levels of 31 infants (group 1) were 23.6 mAU/mL (standard deviation [SD] 5.8), showing extremely low levels, and close to healthy adult levels, with a smaller deviation than what was seen in group 2. The levels of the 46 infants in group 2 were 27.8 (SD 16.0). This does not differ significantly from group 1, but a small number of infants showed a modestly high level in PIVKA-II. There also was no significant difference between the two groups in the HPT. These data would indicate that maternal vitamin K supplementation can maintain the vitamin K status throughout the late neonatal period and prevent an onset of vitamin K-deficient hemorrhage. PMID- 12536347 TI - Prothrombin levels in newborn infants by the carinactivase-1 method. AB - A recently developed method to quantitate prothrombin in plasma uses the carinactivase-1 (CA-1) method. The present study was designed to establish the reference value by the CA-1 method in the neonatal period and to explore the effect of gestational age, birth weight, concurrent diseases, and vitamin K administration on the prothrombin levels. We enrolled 78 consecutive neonates. The gestational ages ranged from 28 to 41 weeks, and the birth weight ranged from 850 to 3750 g. Twenty-nine infants had concurrent diseases, and the others (n = 49) were healthy. A 300 microL blood sample was drawn into a plastic syringe containing 60 U freeze-dried buffered heparin. Prothrombin levels did not differ between appropriate-for-date (AFD) and light-for-date (LFD) babies (p = 0.090) or between groups with and without concurrent diseases (p = 0.210). In healthy AFD babies, prothrombin levels correlated with gestational age (r = 0.465, p = 0.003) and birth weight (r = 0.458, p = 0.003). In healthy low-birth-weight infants (n = 14) and those with concurrent diseases (n = 17), the changes after vitamin K administration were not significant. The CA-1 method is of clinical use in monitoring coagulation during the early neonatal period. PMID- 12536348 TI - Increased expression of plasminogen activator inhibitor-1 with fibrin deposition in a murine model of aging, "Klotho" mouse. AB - Although aging accompanies specific pathological changes, including thrombosis and organ sclerosis, the underlying mechanisms of these processes remain to be elucidated. In the present study, we analyzed the gene expression of plasminogen activator inhibitor-1 (PAI-1), a key molecule in the development of thrombosis, in a murine model of aging, klotho mutant ( kl/kl) mice. Active PAI-1 antigen in plasma and PAI-1 mRNA in several tissues were strikingly elevated in kl/kl mice as compared with wild-type mice. This increased PAI-1 expression was age dependent and linked to the development of ectopic calcification and glomerular fibrin deposition in the kidneys. In situ hybridization analysis of kl/kl mice demonstrated that strong signals for PAI-1 mRNA were localized in renal tubular epithelial cells, cardiomyocytes, adrenal medullar cells, and smooth muscle and endothelial cells in Monckeberg's arteriosclerotic vessels. Renal glomerular fibrin deposition, as evaluated immunohistochemically, was occasionally observed only in kl/kl mice, and the number of fibrin-positive glomeruli increased as the kl/kl mice aged. These observations suggest that in the process of aging the PAI 1 gene expression is increased, contributing to the development of thrombosis. PMID- 12536349 TI - Aging and thrombosis. AB - Advanced age is associated with a dramatic increase in the rates of venous and arterial thrombotic events. Increases in fibrinogen, factors VIII and IX, and other coagulation proteins, without a proportional increase in anticoagulant factors, likely contribute to this risk. Recent studies have delineated a role for genomic elements in controlling age-related expression of some coagulation proteins. Enhanced platelet activity as well as molecular and anatomic changes in the vessel wall also contribute to the thrombotic propensity. Advanced age is associated with elevated interleukin-6 (IL-6) and C-reactive protein levels, indicating an inflammatory state that may be an important stimulus for thrombus formation in the elderly. Despite evidence of a prothrombotic state, many elderly people do not experience clinical thrombotic events. It is possible that the increase in coagulation proteins and activation markers conveys a survival advantage, such as inhibiting tumor angiogenesis. The recent epidemic in obesity may heighten thrombotic risks in the elderly because adipose tissue is an important source of inflammatory cytokines and plasminogen activator inhibitor-1 (PAI-1). As the population ages, further studies will be warranted to define the mechanisms for thrombosis in the elderly. PMID- 12536350 TI - Thromboembolic disease in surgery for malignancy-rationale for prolonged thromboprophylaxis. AB - Patients undergoing surgery for malignancy are at increased risk of initial and recurrent venous thromboembolism (VTE). Several factors have been found to increase the risk of deep vein thrombosis (DVT) in cancer patients both during the first days after the operation and after discharge from hospital. Although, in general, thromboprophylaxis is provided to cancer patients after surgery, the length of time these patients require prophylaxis has not yet been established. Autopsy series, clinical series, and clinical trials indicate that up to about 40% of VTE occurs post discharge. General surgical patients undergoing major abdominal surgery require VTE prophylaxis, and prolonged thromboprophylaxis should be considered in the post-discharge period in high-risk patients, particularly those with cancer. Evidence from studies in general and orthopedic surgery show that prolonged prophylaxis reduces the number of thromboembolic events after discharge from hospital. Prophylaxis should be simple, safe, and effective and should be administered easily to allow continuation of therapy after discharge. Low-molecular-weight heparins are potentially the most suitable agents for long-term thromboprophylaxis in cancer patients. PMID- 12536351 TI - Venous thromboembolic risk and its prevention in hospitalized medical patients. AB - In medical patients, the risk of venous thromboembolism (VTE) is substantially underestimated and prophylaxis is used far less than in surgical patients, reflecting the scarcity of evidence supporting antithrombotic therapy in nonsurgical settings. However, current consensus documents recommend assessment of all medical, as well as surgical, patients for thromboembolic risk and provide prophylaxis recommendations according to the risk level, determined by the presence of different clinical and molecular risk factors. Although long-term, underlying clinical and molecular risk factors also have a major impact on overall risk in medical patients; risk clearly varies with the type of medical condition. Myocardial infarction, stroke, and malignant disease are linked to a high rate of VTE; recent evidence highlights patients with cardiopulmonary disease as a distinct risk group. Despite skepticism in some quarters, high quality evidence confirms the efficacy of unfractionated heparin (UFH) and low molecular-weight heparins (LMWH) in reducing asymptomatic deep vein thrombosis (DVT) in a broad spectrum of medical patients; further studies are required to clarify the effect on fatal pulmonary embolism (PE). Emerging data from the recent PRINCE and MEDENOX studies demonstrate that the LMWH enoxaparin provides effective and well-tolerated VTE prevention in patients with severe cardiopulmonary disease. Emerging evidence has led to a grade 1A recommendation for the use of thromboprophylaxis in these patients in the most recent consensus conference on thromboprophylaxis. Further studies, however, are required to clarify the optimal duration of prophylaxis in medical patients and to evaluate the potential role of outpatient prophylaxis. Accurate risk assessment and prompt implementation of appropriate prophylaxis, selected on the basis of evidence from well-designed controlled clinical trials, may reduce the future morbidity and mortality due to VTE in medical patients. PMID- 12536355 TI - Regulation of gonadotropin secretion: implications for polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is a disorder characterized by hyperandrogenism and chronic anovulation. Although the etiology of PCOS is unknown, perturbations of gonadotropin secretion are one of the hallmarks of this disorder. In normal menstrual physiology, the monotropic rise of plasma follicle-stimulating hormone (FSH) during the luteal-follicular transition is critical for follicular development and subsequent ovulation. One of the mechanisms by which FSH is differentially synthesized involves the luteal slowing of gonadotropin-releasing hormone (GnRH) pulse frequency by ovarian steroids. In PCOS, plasma leutinizing hormone (LH) is commonly increased, FSH is typically in the lower follicular range, and LH (and by inference GnRH) pulse frequency is persistently rapid at approximately one LH pulse per hour. The etiology of the neuroendocrine abnormalities in PCOS remain unclear; however, recent studies have revealed decreased sensitivity of the GnRH pulse generator to inhibition by ovarian steroids, particularly progesterone. This abnormality is reversed by the androgen receptor antagonist flutamide, suggesting that elevated androgen levels may alter the sensitivity of the hypothalamic GnRH pulse generator to steroid inhibition and lead to enhanced LH secretion. As such, women with PCOS require higher levels of progesterone to slow the frequency of GnRH pulse secretion, resulting in inadequate FSH synthesis and persistent LH stimulation of ovarian androgens. The decreased sensitivity of the GnRH pulse generator may help to explain the genesis of PCOS during puberty. In normal early puberty, sleep-entrained increases in LH stimulate ovarian steroids, which subsequently suppress LH frequency and amplitude during the subsequent day. In hyperandrogenemic girls destined to develop PCOS, this nocturnal increase in ovarian steroids may not be adequate to suppress the GnRH pulse generator, leading to a persistently rapid LH pulse frequency, impaired FSH production, and inadequate follicular development. PMID- 12536357 TI - Gonadotropinomas. AB - Advances in immunocytochemistry, electron microscopy, cell culture, and molecular techniques have demonstrated that 80 to 90% of the clinically nonfunctioning pituitary adenomas are gonadotrope-derived and recently recognized as gonadotropinomas, which account for as many as 40 to 50% of all pituitary macroadenomas. Patients usually present with mass effects including visual field loss and headache, hypogonadism, and hypopituitarism. Commonly, the tumor is found incidentally. Recently, a few patients with gonadotropinomas were reported to have hormonal hypersecretion syndromes such as ovarian hyperstimulation, testicular enlargement, and precocious puberty. The tumors can be divided into two broad categories: functioning gonadotropinomas with positive immunostaining for follicle-stimulating hormone, leutinizing hormone, and/or their subunits; and nonfunctioning gonadotropinomas or null cell tumors with negative immunostaining for all pituitary hormones but positive nuclear immunostaining for steroid factor 1 or DAX-1 characteristic of gonadotrope differentiation, with evidence of gonadotropin production or gene expression at the mRNA level. Gonadotropinomas are monoclonal in origin but the pathogenesis of these tumors is unknown and factors that stimulate clonal proliferation not yet determined. A new pituitary oncogene, pituitary tumor transforming gene, has recently been found to be overexpressed in about two thirds of these tumors but it is also detected in all other pituitary tumor subtypes. Alterations of tumor hormone receptors and local growth factors may also play a role in the tumor development and/or progression. Transphenoidal surgery remains the principal therapy for the macroadenomas. Radiosurgery using gamma knife, the linear accelerator, or proton beam therapy showed promising results, especially for controlling the residual or recurrent tumors. Medical therapy with somatostatin analogs, dopamine agonists, and gonadotropin-releasing hormone agonists and antagonists are rarely effective in reducing tumor size. Experimental therapy with intraoperative local chemotherapy or potential gene therapy requires further investigation. PMID- 12536356 TI - Hypogonadotropic hypogonadism. AB - Hypogonadotropic hypogonadism is characterized by failure of gonadal function secondary to deficient gonadotropin secretion, resulting from either a pituitary or hypothalamic defect, and is commonly seen in association with structural lesions or functional defects affecting this region. Although the genetic basis for idiopathic hypogonadotropic hypogonadism is largely unknown, mutations in several genes involved in the hypothalamo-pituitary-gonadal axis development and function have recently been implicated in the pathogenesis of this condition. Genes currently recognized to be involved include KAL-1 (associated with X-linked Kallmann Syndrome), gonadotropin-releasing hormone (GnRH) receptor, gonadotropins, pituitary transcription factors (HESX1, LHX3, and PROP-1), orphan nuclear receptors (DAX-1, associated with X-linked adrenal hypoplasia congenital, and SF-1), and three genes also associated with obesity (leptin, leptin receptor, and prohormone convertase 1 [ PC1]). Study of these mutations provides an important contribution in the understanding of the different stages of the reproductive axis development and physiology. Treatment options currently available for puberty induction, maintenance replacement therapy, and fertility induction are considered here. Gametogenesis can be induced with either exogenous gonadotropin or pulsatile GnRH therapy, depending on the etiology. PMID- 12536358 TI - GnRH agonists, antagonists, and assisted conception. AB - Pituitary stimulation with pulsatile gonadotropin-releasing hormone (GnRH) induces both follicle-stimulating hormone and luteinizing hormone (LH). Blockade of pituitary gonadotropin secretion occurs upon desensitization when a continuous GnRH stimulus is provided by means an agonist or when the pituitary receptors are occupied with a competitive antagonist. The most common indication for blockade of pituitary gonadotropin secretion is with assisted reproduction treatment (ART) where it prevents premature luteinization. Originally by lack of clinically available GnRH antagonist, prolonged daily injection of agonist with its desensitizing effect was introduced for this purpose. Today, single- and multiple dose injectable antagonists are also available to block the LH surge. This review provides an overview of the use of GnRH agonists and antagonists in assisted reproduction. PMID- 12536359 TI - Hyperprolactinemia: etiology, diagnosis, and management. AB - Hyperprolactinemia is the most common endocrine disorder of the hypothalamic pituitary axis. A prolactinoma is the most common cause of chronic hyperprolactinemia once pregnancy, primary hypothyroidism, and drugs that elevate serum prolactin levels have been excluded. Patients can present with hypogonadism, infertility, galactorrhea, osteopenia, and mass effects of the tumor. When hyperprolactinemia is confirmed, a cause for the disorder needs to be sought. This involves a careful history and examination, followed by laboratory tests and diagnostic imaging of the sella turcica. The goals of treatment are to normalize prolactin levels, restore gonadal function, and reduce the effects of chronic hyperprolactinemia. Dopamine agonists are the treatment of choice for the majority of patients. Transsphenoidal surgery is usually reserved for patients who are intolerant of or resistant to dopamine agonists or when hyperprolactinemia is caused by non-prolactin-secreting tumors compressing the pituitary stalk. Cabergoline has been shown to be more effective and better tolerated than bromocriptine. However, there are more data on the safety of the latter drug during pregnancy and bromocriptine, therefore, remains the treatment of choice in hyperprolactinemic women wishing to conceive. PMID- 12536360 TI - Corticosteroids and pregnancy. AB - Pregnancy results in major changes in the hypothalamo-pituitary-adrenal (HPA) axis, which in turn influence fetal growth and the timing of labor. From the beginning of the second trimester maternal cortisol secretion increases, and in late pregnancy the placenta, in large part mediated through corticotroph releasing hormone, plays a crucial role in the regulation of the fetal HPA axis to ensure the synchronization of the various processes involved in parturition. Exposure of the fetus to excess glucocorticoid results in intrauterine growth failure and possibly "programs" the development of cardiovascular disease in adult life. Biochemical assessment of the HPA axis is complicated by the estrogen induced elevation of circulating cortisol-binding globulin, resulting in misleadingly high circulating cortisol levels The hypercortisolemia of Cushing's syndrome causes infertility, but if pregnancy does occur it can result in increased morbidity and mortality in mother and fetus. However, the prospects of a successful pregnancy are greatly improved with control of hypercortisolemia by surgery and medical therapy with metyrapone. Hypoadrenalism can be difficult to diagnose during pregnancy but, once the diagnosis is made, with careful monitoring, dose adjustment as indicated, and parenteral cover for labor, a successful pregnancy should result. PMID- 12536361 TI - Thyroid dysfunction: reproduction and postpartum thyroiditis. AB - Thyroid function during pregnancy is characterized by changes in circulating thyroid hormone concentrations related to alterations in thyroxine binding globulin (TBG), human chorionic gonadotropin (hCG), and iodine status. The immunology of normal pregnancy shows a reduction in antibody titer during gestation and an increase in T helper-2 (TH2) immune responses. Thyroid dysfunction may cause menstrual disturbances in hyper- and hypothyroidism but less marked disturbances of sexual function in men. Fertility is reduced in hypo- and hyperthyroid females. Accumulating evidence suggests a strong association between the presence of thyroid antibodies and fetal loss, although the data relating to recurrent abortion are not so convincing. Asymptomatic maternal gestational hypothyroidism may occur in up to 2.5% of women; studies have shown a significant impact of this condition in causing a decrease of child IQ, suggesting that screening for maternal hypothyroidism with intervention may be justified. Postpartum thyroid disease occurs in 5 to 9% of women and thyroid dysfunction postpartum is seen in 50% of thyroid peroxidase antibody positive (TPO Ab+ve) women. There is a significant rate of hypothyroidism in long-term follow-up of women who have transient postpartum thyroid dysfunction. PMID- 12536362 TI - Appetite control and reproduction: leptin and beyond. AB - It is now recognized that appropriate regulation of reproduction, energy intake, and energy expenditure, and thus maintenance of body weight and fertility, relies on complex hypothalamic neurocircuitry. Feeding and reproductive function are closely linked. During times of undernourishment and falling body fat the reproductive axis is down-regulated. Circulating factors and hypothalamic circuits coordinate these responses. Leptin has been described to be an important peripheral signal that indicates body fat stores to the hypothalamus and thus links nutrition and reproduction. Leptin acts by altering neuropeptide circuits in the hypothalamus, which alter gonadotropin-releasing hormone release and food intake. The importance of key neuropeptide systems identified in rodents is now being established in man. Notably mutations in the melanocortin MC4 receptor are found in up to 4% of the morbidly obese, and in a proportion of patients with anorexia nervosa mutations have been identified in the agouti-related peptide gene (AgRP), which codes for an endogenous antagonist of this receptor. Intranasal administration of a melanocortin fragment known to activate the MC4 receptor decreases adiposity in humans. The melanocortin system has been shown to influence the reproductive axis in rodents. However, the role of the melanocortin system in the control of reproduction in humans remains to be established. Since the discovery of leptin, attention has also been focused on peripheral signals that regulate reproduction, food intake, and energy expenditure, either directly or via feedback on hypothalamic circuits. Notable new discoveries in this area include the gastric hormone ghrelin. Circulating ghrelin stimulates food intake in rodents and humans, although an influence on the reproductive axis is yet to be reported. PMID- 12536363 TI - White matter disease in cerebral organic acid disorders: clinical implications and suggested pathomechanisms. AB - White matter abnormalities (including dys-, hypo-, demyelination and delayed myelation) are frequently found in cerebral organic acid disorders, a recently delineated subgroup of inherited organic acid disorders presenting predominantly with a neurological symptomatology. Biochemically, this disease subgroup lacks metabolic derangements, such as hypoglycemia, hyperammonemia and acidosis, and is characterized by an accumulation of organic acids that share structural similarities with the excitatory amino acid glutamate (D-2-, L-2-, 3 hydroxyglutarate, glutarate) or have been suggested as neurotransmitters/neuromodulators ( N-acetylaspartylglutamate). Evidence from in vitro and in vivo studies is growing that relevant organic acids significantly contribute to the neuropathology of these diseases via interference with glutamatergic or GABAergic neurotransmission or impairment of energy metabolism. This article provides an overview on the clinical and neuroradiological presentation of white matter disease in cerebral organic acid disorders, focusing on the suggested pathomechanistic relevance of excitotoxicity, oxidative stress, and impaired energy metabolism. PMID- 12536364 TI - Congenital creatine transporter deficiency. AB - BACKGROUND: Two inborn errors of metabolism of creatine synthesis as well as the X-linked creatine transporter (SLC6A8) deficiency have been recognized. This report describes the features of five identified male patients and their female relatives who are carriers of the X-linked creatine transporter deficiency syndrome. METHODS: Proton MR spectroscopy was used to recognize creatine deficiency in the patients. Molecular analysis of the SLC6A8 gene was performed, confirming the diagnosis of homozygous males and heterozygous females. RESULTS: We describe four families from a metropolitan area in the U. S. with X-linked creatine transporter deficiency. All affected males present with developmental delay and severe developmental language impairment. Proton MR spectroscopy shows significantly depressed to essentially absent creatine and phosphocreatine in the male patients. Nonsense mutations and amino acid deletions were found in the SLC6A8 gene in the affected families. CONCLUSION: Creatine transporter deficiency may be a more common X-linked genetic disorder than originally presumed. The affected males exhibit mental retardation with severe expressive language impairment. PMID- 12536365 TI - Relationship between head circumference and brain volume in healthy normal toddlers, children, and adults. AB - OBJECTIVE: To quantify the relationship between brain volume and head circumference from early childhood to adulthood, and quantify how this relationship changes with age. METHODS: Whole-brain volume and head circumference measures were obtained from MR images of 76 healthy normal males aged 1.7 to 42 years. RESULTS: Across early childhood, brain volume and head circumference both increase, but from adolescence onward brain volume decreases while head circumference does not. Because of such changing relationships between brain volume and head circumference with age, a given head circumference was associated with a wide range of brain volumes. However, when grouped appropriately by age, head circumference was shown to accurately predict brain volume. Head circumference was an excellent prediction of brain volume in 1.7 to 6 years old children (r = 0.93), but only an adequate predictor in 7 to 42 year olds. CONCLUSIONS: To use head circumference as an accurate indication of abnormal brain volume in the clinic or research setting, the patient's age must be taken into account. With knowledge of age-dependent head circumference-to-brain volume relationship, head circumference (particularly in young children) can be an accurate, rapid, and inexpensive indication of normalcy of brain size and growth in a clinical setting. PMID- 12536366 TI - Neurological and perceptual-motor outcome at 5 - 6 years of age in children with neonatal encephalopathy: relationship with neonatal brain MRI. AB - OBJECTIVE: The aims of this study were 1) to determine the incidence of minor neurological dysfunction and perceptual-motor difficulties in children aged 5-1/2 -- 6-1/2, who had been born full-term but presented with neonatal encephalopathy (NE) and low Apgar scores and 2) to examine the relationships between the presence/absence of these difficulties with neonatal brain MRI. PARTICIPANTS AND METHODS: Sixty-eight full-term infants with one minute Apgar scores less than or equal to 5 and neurological abnormalities during the first 48 hours after birth were included in the study. All children had a neonatal MRI brain scan. Surviving infants were assessed between the age of 5 and 6 years using the Touwen Examination, the Movement ABC and the WPPSI-R. RESULTS: Fifteen of the 68 infants (22 %) died in the neonatal period. Of the 53 surviving infants, 19 (36 %) had cerebral palsy. The remaining 34 were considered normal at 2 years of age but, when assessed at school age, 8 (15 %) had minor neurological dysfunction and/or perceptual-motor difficulties, 1 (2 %) had only cognitive impairment and 25 (47 %) were normal. The outcome largely reflected the pattern of lesions on brain imaging. While 83 % of those with a normal outcome had normal scans or minimal white matter lesions, 80 % of those with minor neurological dysfunction and/or perceptual-motor difficulties had mild or moderate basal ganglia or more marked white matter lesions. CONCLUSION: Continued surveillance is recommended for children with apparently normal outcome at two years of age after NE, particularly when abnormalities are detected on brain MRI. PMID- 12536367 TI - Congenital myasthenic syndrome (CMS) in three European kinships due to a novel splice mutation (IVS7 - 2 A/G) in the epsilon acetylcholine receptor (AChR) subunit gene. AB - Mutations in the epsilon-acetylcholine receptor (AChR epsilon) subunit gene cause congenital myasthenic syndromes (CMS) with postsynaptic neural transmission defects. We present 3 male and 2 female patients from three unrelated Croatian, Hungarian, and Russian families with autosomal recessive CMS. All patients manifested with variable degrees of ophthalmoparesis and generalized, fatiguable muscle weakness since birth or early infancy. Electrophysiological studies showed a decremental response in all patients indicating a neuromuscular transmission defect. Pyridostigmine treatment improved the proximal muscle weakness whereas the ophthalmoparesis remained unchanged in all patients. Analysis of the AChR epsilon subunit gene showed homozygosity for a novel splice site mutation of intron 7 epsilon(IVS7-2A/G) in the two Croatian siblings. epsilon-mRNA analysis by RT-PCR and direct sequencing revealed that exon 7 was spliced directly to exon 9 with skipping of exon 8. The Hungarian and Russian patients were heteroallelic carriers of the same mutation epsilon(IVS7-2A/G) and of a frameshifting mutation epsilon 70insG and epsilon 1293insG, respectively. We hypothesize that altered splice products may not be expressed as functional receptors at the cell surface. A haplotype analysis with polymorphic markers revealed a high degree of similarity for the epsilon(IVS7-2A/G) carrying allele in all families and may therefore indicate a common origin of the mutation. PMID- 12536368 TI - Epilepsy surgery outcome in children with focal epilepsy due to tuberous sclerosis complex. AB - Tuberous sclerosis complex (TSC) is frequently associated with focal epilepsy due to cerebral tubers. Seizures are the first symptoms in most patients with brain involvement. These epilepsies are frequently severe, drug-resistant and may have a negative impact on the child's global development. Although most epilepsies are multicentric, these patients may be candidates for epilepsy surgery, if it is possible to determine a leading epileptogenic tuber. Nine patients with TSC were examined with long-term video-EEG monitoring, different neuroimaging techniques and neuropsychological tests. A main epileptogenic tuber could be identified in all of our patients. We found good correlations between neuroimaging and EEG. Surgery was performed in eight patients. Seizure outcome was good in all. Two patients became seizure-free, one patient had a single prolonged seizure five days postoperatively, four patients had a significant seizure reduction of more than 75 % and one patient had a seizure reduction of more than 50 % after surgery without additional neurologic deficits. In conclusion, patients with TSC and drug resistant epilepsy may benefit from epilepsy surgery with reduction in frequency and severity of seizures as well as improved mental and behavioural development leading to a better quality of life. In view of recent developments in functional and metabolic imaging, primary epileptogenic lesions will be more easily detectable in patients with diffuse brain involvement in TSC and surgical treatment may be more specifically applied at an earlier age to a selected subgroup of patients with this disorder. PMID- 12536369 TI - Hypercytokinemia in hemiconvulsions-hemiplegia syndrome associated with dual infection with varicella zoster and Epstein-Barr viruses. AB - Hemiconvulsions-hemiplegia (HH) syndrome is an acquired condition in which hemiplegia develops after a preceding febrile unilateral status epilepticus in a previously healthy child. Although viral encephalitis or vascular diseases may be the underlying etiology, the pathogenesis remains unknown in the majority of cases. We measured both plasma and cerebrospinal fluid cytokine levels in a girl with HH syndrome, and found elevated plasma concentrations of soluble interleukin 2 receptor and tumor necrosis factor-alpha, and a slightly increased plasma level of interleukin-6. Furthermore, she had a high serum concentration of soluble E selectin, which is a marker of inflammatory endothelial activation. These findings suggest that proinflammatory cytokine-induced cerebrovascular endothelial injury could play a role in the pathogenesis of HH syndrome. PMID- 12536370 TI - A case of diffuse leptomeningeal oligodendrogliomatosis associated with HHV-6 variant A. AB - We describe a rare case of diffuse leptomeningeal oligodendrogliomatosis associated with the human herpes virus 6 variant A (HHV-6 A). A 2-year-old boy presented with progressive neurological symptoms and hydrocephalus. The patient had a VP shunt placement but did not fully recover. HHV-6 A was detected in both CSF and serum by nested PCR. His symptoms improved repeatedly, but temporarily, on antiviral treatment. An open brain biopsy, ten months after presentation, revealed leptomeningeal tumour as well as the presence of viral DNA in the tumour tissue. The role of HHV-6 A could be that of a reactivated opportunist. However, this case also raises the question whether this neurotropic virus, with malignant transforming properties in vitro, may have a role in pathogenesis in some cases of brain malignancy. PMID- 12536371 TI - Hypothalamic growth hormone deficiency and supplementary GH therapy in two patients with mitochondrial myopathy, encephalopathy, lactic acidosis and stroke like episodes. AB - Two pediatric patients with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes were diagnosed with growth hormone deficiency with the primary lesion identified as the growth hormone-releasing factor producing cells of the hypothalamus. Stimulation tests with insulin, levodopa and sleep did not overcome the deficient pattern of growth hormone secretion. By comparison, the growth hormone-releasing factor stimulation test generated a normal growth hormone response in these two patients. Growth hormone supplementary therapy was effective in terms of growth gain without adverse effects. PMID- 12536372 TI - Severe infantile measles encephalitis occurred three months after neonatal measles. AB - A 3-month-old girl who developed severe measles encephalitis after neonatal measles is reported. Her mother had measles when she was ten days old and she was admitted to our hospital with low grade fever, Koplik spot, and mild exanthema seventeen days after birth, and she recovered in 7 days without any complication. At three months of age, she was readmitted because of intractable seizures. The levels of IgM and IgG antibodies against measles in the cerebrospinal fluid were elevated. The measles virus genome, amplifying the region encoding the nucleocapsid protein, was detected from the brain specimen by reverse transcriptase-polymerase chain reaction. Magnetic resonance imaging showed a focal destructive lesion and diffuse cerebral atrophy. The electroencephalogram did not show periodic synchronous discharges. Although the neonatal measles was believed to be relatively mild in severity, the possible development of measles encephalitis should be carefully monitored in an infant who had neonatal measles. PMID- 12536374 TI - Visualization of molecular and cellular events with green fluorescent proteins in developing embryos: a review. AB - During the past 5 years, green fluorescent protein (GFP) has become one of the most widely used in vivo protein markers for studying a number of different molecular processes during development, such as promoter activation, gene expression, protein trafficking and cell lineage determination. GFP fluorescence allows observation of dynamic developmental processes in real time, in both transiently and stably transformed cells, as well as in live embryos. In this review, we include the most up-to-date use of GFP during embryonic development and point out the unique contribution of GFP visualization, which resulted in novel discoveries. PMID- 12536373 TI - The genetic heritage of the earliest settlers persists both in Indian tribal and caste populations. AB - Two tribal groups from southern India--the Chenchus and Koyas--were analyzed for variation in mitochondrial DNA (mtDNA), the Y chromosome, and one autosomal locus and were compared with six caste groups from different parts of India, as well as with western and central Asians. In mtDNA phylogenetic analyses, the Chenchus and Koyas coalesce at Indian-specific branches of haplogroups M and N that cover populations of different social rank from all over the subcontinent. Coalescence times suggest early late Pleistocene settlement of southern Asia and suggest that there has not been total replacement of these settlers by later migrations. H, L, and R2 are the major Indian Y-chromosomal haplogroups that occur both in castes and in tribal populations and are rarely found outside the subcontinent. Haplogroup R1a, previously associated with the putative Indo-Aryan invasion, was found at its highest frequency in Punjab but also at a relatively high frequency (26%) in the Chenchu tribe. This finding, together with the higher R1a-associated short tandem repeat diversity in India and Iran compared with Europe and central Asia, suggests that southern and western Asia might be the source of this haplogroup. Haplotype frequencies of the MX1 locus of chromosome 21 distinguish Koyas and Chenchus, along with Indian caste groups, from European and eastern Asian populations. Taken together, these results show that Indian tribal and caste populations derive largely from the same genetic heritage of Pleistocene southern and western Asians and have received limited gene flow from external regions since the Holocene. The phylogeography of the primal mtDNA and Y chromosome founders suggests that these southern Asian Pleistocene coastal settlers from Africa would have provided the inocula for the subsequent differentiation of the distinctive eastern and western Eurasian gene pools. PMID- 12536375 TI - Rapid determination of rice seed vigour by spontaneous chemiluminescence and singlet oxygen generation during early imbibition. AB - Using a highly sensitive single photon counter, a spontaneous chemiluminescence (CL) study on rice (Oryza sativa L.) seeds stored in different years was carried out. We first observed that the degree of ageing in rice seeds was related to the intensity of spontaneous CL during early imbibition (0-30 min). Rice seeds stored for a shorter time had a stronger intensity of CL in early imbibition. The germination rate of rice seeds showed an obvious positive correlation with the intensity of spontaneous CL. Singlet oxygen ((1)O(2)) in rice seeds during early imbibition was investigated by a CL method using a cypridina luciferin analogue, 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo [1,2alpha] pyrazin-3-one (MCLA), as a selective CL probe. Additional experimental evidence for the formation of (1)O(2) came from the quenching effect of sodium azide (NaN(3)) on MCLA-mediated rice seeds' CL. Analysis based on the experimental results demonstrated that spontaneous CL in rice seeds during early imbibition was mainly contributed by singlet oxygen ((1)O(2)). PMID- 12536376 TI - Analysis of several fluorescent detector molecules for protein microarray use. AB - The utility of several streptavidin-linked fluorescent detector molecules was evaluated on two protein microarray platforms. Tested detector molecules included: Alexa Fluor 546; R-phycoerythrin (RPE), orange fluospheres; Cy3 containing liposomes (Large Unilamellar Vesicles, LUV) labelled with Cy3; and an RPE-antibody complex. The two array architectures tested consisted of an array of murine Fc-biotin and an array of murine IgG (the murine IgG array was probed with a biotinylated rabbit anti-murine IgG). These platforms allowed for the direct comparison of detector utility by detector recognition of array-bound biotin. All of the fluorescent detectors examined demonstrated utility on each of the array platforms. For the Fc-biotin array, detector signal intensity (background adjusted) was as follows: RPE-antibody complex > fluospheres > RPE > liposomes > Alexa 546: for the IgG array: RPE/antibody complex > RPE > fluospheres > Alexa546 > liposomes. The RPE-antibody complex fluoresced 67% and 150% more intensely than the next closest detector molecule for the Fc-biotin and the murine IgG arrays, respectively. A marked increase in background fluorescence (as compared to RPE alone) did not accompany the increase in signal intensity gained through RPE antibody complex use (a true increase in signal:noise ratio). These results suggest that the RPE-antibody complex is superior to other molecules for fluorescent detection of analytes on protein microarrays. PMID- 12536377 TI - Determination of phenol using an enhanced chemiluminescent assay. AB - Enhanced chemiluminescence (ECL) describes the phenomenon of increased light output in the luminol oxidation reaction catalysed by horseradish peroxidase (HRP) in the presence of certain compounds, such as para-iodophenol. In this work, the effects of phenol on the para-iodophenol-enhanced HRP-catalysed chemiluninescent reaction intensity in an aqueous buffer (Tris-HCl buffer, pH 8.5) and in a surfactant-water-octane mixture were compared. Preincubation of HRP at low phenol concentrations stimulated the chemiluminescent intensity in the assay performed in an aqueous buffer, but did not have significant effect in the sodium bis(2-ethylhexyl)sulphosuccinate) (Aerosol OT, AOT) applied system. It was also observed that HRP preincubation with phenol concentration higher than 0.003 mg/mL produced an inhibitory effect on the enzyme activity for both assay systems. Only an inhibitory effect of phenol on the chemiluminescent intensity in the surfactant system in octane (as organic solvent) was observed. Three assays were developed to determine phenol concentration in water and in an organic solvent mixture. The detection limits were 0.006, 0.003 and 0.0005 mg/mL, respectively, for the buffer-containing system, the AOT-applied system with phenol standard solutions in water and for the AOT-applied system with phenol standard solutions in octane. PMID- 12536378 TI - Imaging of ultra-weak bio-chemiluminescence and singlet oxygen generation in germinating soybean in response to wounding. AB - Ultra-weak bio-chemiluminescence (UBC) from germinating soybean (Glycine max L. Merr) cotyledon under mechanical wounding was observed using a high-sensitivity imaging system based on an ICCD detector and a highly sensitive single photon counter (SPC) device. The UBC imaging showed that the intensity at the injury location on a wounded cotyledon was obviously enhanced as compared with that at the non-injured point. The UBC intensity of wounded cotyledons was initially very high and reached a stationary state after about 5 min. Wounding-induced emission could be suppressed by wounding in the presence of sodium azide. Deuterium oxide amplified the emission intensity. It was concluded that singlet oxygen ((1)O(2)) was the main cause of the emission during the wounding phase. PMID- 12536379 TI - Studies of visible oscillating chemiluminescence with a luminol-H2O2-KSCN-CuSO4 NaOH system in batch reactor. AB - Oscillating chemical reactions are complex systems involving a large number of chemical species. In oscillating chemical reactions some species, usually reaction intermediates, exhibit fluctuation in concentration. Visible oscillating chemiluminescence, produced by the addition of luminol (3-aminophthalhydrazide) to the oscillating system H(2)O(2)-KSCN-CuSO(4)-NaOH, was investigated. In this study the effect of varying the concentration of H(2)O(2), KSCN, CuSO(4), NaOH and luminol was investigated in a batch reactor. We showed that the concentration of all components involved in the oscillating chemilumenscent reaction influenced the light intensity and the oscillation period. PMID- 12536380 TI - Chemiluminescence of lucigenin by electrogenerated superoxide ions in aqueous solutions. AB - The chemiluminescence reaction of lucigenin (Luc(2+)c2NO(3) (-), N,N'-dimethyl 9,9'-biacridinium dinitrate) at gold electrodes in dioxygen-saturated alkaline aqueous solutions (pH 10) was investigated in detail by the use of electrochemical emission spectroscopy. We noted that both O(2) and Luc(2+) are reduced on a gold electrode in aqueous solution of pH 10 in almost the same potential region. From this fact, we expected chemiluminescence based on a radical-radical coupling reaction of superoxide ion (O(2).(-)) and one-electron reduced form of Luc(2+) (Luc.(+), a radical cation). Chemiluminescence was actually observed in the potential range where O(2) and Luc(2+) were simultaneously reduced at the electrodes. The effects were examined upon addition of enzymes, i.e. superoxide dismutase (SOD) and catalase, into the solution and the substitution of heavy water (D(2)O) for light water (H(2)O) as a solvent on the chemiluminescence. In the presence of native and active SOD, chemiluminescence was completely absent. On the other hand, chemiluminescence was observed, unchanged in the presence of either denatured and inert SOD or catalase. In addition, the amount of chemiluminescence in D(2)O solution was about three times greater than that in H(2)O solution. These results, together with cyclic voltammetric results, suggest that O(2).(-) participates directly in the chemiluminescence but H(2)O(2) does not, and the chemiluminescence results from the coupling reaction between O(2).(-) and Luc(.+) under the present experimental conditions. These chemically unstable species, O(2).(-) and Luc.(+), are produced during the simultaneous electroreduction of O(2) and Luc(2+). The coupling reaction between those radical species would lead to the formation of a dioxetane-type intermediate and, finally, to chemiluminescence. The chemiluminescence reaction mechanism is discussed. PMID- 12536381 TI - Linear polarized near-infrared irradiation stimulates mechanical expansion of the rat sagittal suture. AB - Mechanical sutural separation has been carried out in clinical orthodontics for controlling the growth of the craniofacial skeleton. This study was designed to evaluate the effects of linear polarized near-infrared ray irradiation on the sutural expansion of rat sagittal suture. Twenty 8-week-old Wistar strain male rats were equally divided into experimental and control groups. Suture expansion was carried out for 5 days for all animals using an expansion appliance. The experimental animals were subjected to linear polarized near-infrared irradiation. This study has demonstrated that linear polarized near-infrared irradiation stimulates sutural expansion without any pathological changes. PMID- 12536382 TI - Assessment of training effects on activity levels of alveolar macrophage in matured rats using chemiluminescent technique. AB - Chemiluminescence responses have been used for the evaluation of phagocyte function. In this study, to evaluate effects of training started after maturation on pulmonary immunity, the activity levels of rat alveolar macrophages (AMs) were assessed as reactive oxygen species (ROS) generating capacity, measured by lucigenin- and luminol-dependent chemiluminescence, using a parallel luminometer. One group of male Wistar rats started training at 11 weeks old and another group at 17 weeks old. The experimental period was 12 weeks, and about half of the rats were sacrificed after 6 weeks. The forced and voluntary exercises affect the mean levels of body weights and cell populations in the bronchoalveolar lavage fluid in younger animals; however, the voluntary exercise group in younger animals seemed to adapt after 12 weeks. By contrast, chemiluminescence responses in older rats observed after 6 weeks suggest that AMs are primed, and the maximum releasing activities of ROS are reduced. These changes in AM activity may be caused by the exercise and separation stresses and the rats may adapt to those stressors after 12 weeks. The chemiluminescent technique is thought to be useful to evaluate the changes of AM activity. PMID- 12536383 TI - Forest conservation in the Congo Basin. PMID- 12536384 TI - [Observation of repair of wounded rat skin by affinity histochemical method of SJA]. AB - Time expression of SJA tissue receptors surrounding wounded skin of 40 rats was studied by affinity histochemical method. The results were compared to that by EGFR immunohistochemical method and c-myc situ hydridization method. It was observed that SJA receptors began to decrease after the skin was injured and reached the lowest quantities in 15 minutes. After that, they began to increase and reached the highest quantities in 2 hours. It needs to be further studied whether there is the correlation between SJA receptors and EGFR has the function of repairing wounded skin. PMID- 12536385 TI - [The SEM observation of brain tissues and vascular corrosion cast of closed diffuse brain injuries]. AB - The morphologic changes of the closed injured brain of rats were observed by SEM. The rats either died immediately after conclusive injuries or were killed after 5 minutes to 5 days after injuries. The main changes were as follows: the diffuse disorder, twist, wave-like distortion and break of neuron fibers; axonal swelling; formation of axonal retraction balls; stripping and denotation of myelin sheath The ball-like swelling of neuron, break of neuron membrane and vascular wall, and microthrombus formations were also observed. These damages worsened with prolongation of surviving time of the rats. The axonal retraction ball appeared 8 hours after the injury and was approximately 3-5 cm in diameter, and developed to 7-8 cm after 3-5 days. It is observed that frontal lobe, cerebellum and brain stem were severely damaged. PMID- 12536386 TI - [A pathological study of hemorrhage and necrosis of pancreas due to poisoned death: analysis of 22 cases]. AB - Pathological changes of pancreas in 22 cases died of intoxication were studied and the mechanism was discussed. The results showed that: in death cases by eating of toxicants, the hemorrhage and necrosis of pancreas is extensive mainly in caput and corpus, while inhaling of carbon monoxide is focal in corpus and cauda. PMID- 12536387 TI - [Study of elastic fiber changes of human skin in distinction between antemortem and postmortem wounds]. AB - Elastic fiber changes of volunteers' antemortem and postmortem skin wounds of various time were observed in order to find possible differences between them. These sections of the wounds were stained by Hart's modification of Weigert's elastic tissue stain.. However, this study showed that there were no differences in the nature and distribution of the elastic fibers in the dermis of volunteers' antemortem and postmortem human skin wounds. Therefore, this suggests that the appearances of the elastic fibers in the dermis should not be as a means of differentiating antemortem skin wounds from postmortem wounds in forensic identification. PMID- 12536388 TI - [A preliminary immunohistochemical study of fibronectin in viral myocarditis]. AB - For postmortem diagnosis of viral myocarditis, 12 specimens of heart in autopsy were studied immunohistochemically with anti-fibronectin(FN) antibody. Among 5 cases with definite or suspected myocarditis, intensive FN-positive could be found in all monocytes, macrophages and some neutrophils and there were 3 cases with FN-positive cardiomyocytes. While in 3 violent death cases with leucocytes infiltration in the interstitial tissue of myocardia, no FN-positive cardiomocytes were observed, which was the same as that of 4 case without cardiac pathological changes. Moreover, there were rarely FN-positive leucocytes in the 7 cases without viral myocarditis. The results suggest that the immunohistochemical observation of FN in myocardia might be of value for detecting slight degeneration of cardiomyocytes and determining the inflammatory leucocyte infiltration in myocarditis. PMID- 12536389 TI - [Study of the high polymorphism STR locus-FGA in Chinese population]. AB - Population genetic study of the complicated STR locus-FGA was performed by the method of PCR in Han population samples. A total of 21 different alleles including 7 interalleles and 63 different genotypes were observed in 349 unrelated individuals. The observed genotype discrimination showed no significant deviation from Hardy-Weinberg equilibrium and 51 family studies showed no mutation. According to the results obtained in this study(DP = 0.9612, PE = 0.7007, PIC = 0.8333), this system can be used as an useful means in forensic identification of both criminal and paternity case. PMID- 12536390 TI - [Polymorphism analysis of D19S253 locus in Guangzhou area population]. AB - Frequency data for STR system D19S253 were obtained from 105 unrelated individuals of Guangzhong population. PCR products were detected by horizontal native polyacrylamide gel electrophoresis and a total of 9 alleles were identified by side-by-side comparison with a sequenced allelic ladder prepared by ourselves. The observed genotype distribution conformed with Hardy-Weinberg equilibrium. The high information content found in this system(heterozygosity rate was 0.8353, the mean exclusion chance was 0.7499, the discrimination power was 0.9211, the polymorphism information content was 0.8203) indicated it is a useful means in forensic routine casework both in criminal and paternity cases. PMID- 12536391 TI - [Research of the extraction method of morphine from biological fluids]. AB - A solid-phase extraction method of morphine from urine and blood has introduced. The effect of 5 SPE columns, 3 eluents and pH on morphine recovery has been investigated systematically. Derivative GC was used as a method of detection. The result showed that the column and the eluent of such as GDX-301, GDX-403 and C18 chloroform:isopropanol (9:1) had good behaviors to extraction of morphine. When GDX-301 was used as a sorbent, the recovery of morphine from urine was above 90% at pH 9, then went down with the increase of pH. While the recovery from blood was growing with the increase of pH, which reached above 90% in strong alkaline. The extraction method is simple, inexpensive, efficient and reproducible, which provides an effective and practical method to extract morphine and similar illicit drugs from biological fluids. PMID- 12536392 TI - [Analysis of penicillins in whole blood by reversed high-performance liquid chromatography]. AB - A reversed-HPLC method was established to determinate penicillins in whole blood. Thiamphenicol as internal standard was added to 0.5 ml blood. Proteins in blood were precipitated with acetonitrile, then the separate supernatant was directly injected onto the chromatography column. The range of five penicillins' linearities was 0.9918 to 0.9996, their range of relative recovery was 91.75% to 107.33%, their CV was 3.56% to 9.85%. Effects of different storage temperatures on the stability of analytes and interference of other antibiotics were also studied. PMID- 12536393 TI - [SPE analysis of 4 rodenticides in whole blood and liver by HPLC]. AB - A solid phase extraction (SPE) method has been developed for the analysis of 4 rodenticides: warfarin, couatetralyl, bromadiolone and brodifacoum, using reverse phase high performance liquid chromatograph (HPLC) as determination system. The recovery is 62.6-90.0% (RSD5.1-7.8%) and 37.5-67.7% (RSD5.4%-12.8%) in whole blood and liver sample respectively. The procedure is simple and stable. And the method was confirmed to be effective in the animal experiment. PMID- 12536394 TI - [A study of 44 forensic autopsy cases of adrenal hemorrhage]. AB - 44 cases of adrenal hemorrhage (AH) were analyzed. The forensic autopsy showed that AH occurred mainly in male aged from 20 to 40 years old. The traumatic AH was more often seen in young men and the pathologic AH was mostly seen in elderly persons. AH were more likely found in bilateral adrenal corticomedullary junction. The pathological grade of AH was not related to the interval time of survival. The pathologic mechanism of AH was discussed as well. PMID- 12536395 TI - [Study on assessments of injury grade in 203 cases of traffic accident]. AB - The assessments of victims' injury grade in 203 cases of traffic accidents were reviewed in this article, sorts of these traffic accidents and the best time for assessment of injury grade after traffic accident were also discussed. PMID- 12536396 TI - [The phenomenon of gene linkage and recombination in the paternity test]. AB - The phenomenon of gene linkage and recombination may nearly be overlooked in paternity test of one single child, but it is likely encountered in paternity test of twin or more. In a case of paternity test, the results of 17 items including eight DNA loci were analyzed and the phenomenon of gene linkage and recombination was discussed in detail. This phenomenon should be brought into necessary attention in the paternity test. PMID- 12536397 TI - [The changes of GFAP, PCNA after brain contusion: an immunohistochemical study in forensic pathology]. AB - After the model of experimental brain contusion was set up, the changes of GFAP, PCNA were studied by immunohistochemistry SP method combined with image quantity analysis. After contusion, the gray degrees and areas of GFAP-positive cells increased significantly at the 3rd hour, reached the highest level in the 4th day and kept until the 7th day. PCNA-positive cells appeared in the 12th hour, simultaneously, their gray degrees tended to increase and positive areas tended to decrease. It showed regular changes of GFAP, PCNA with various survival times, and these changes were useful in the diagnosis of brain contusion especially in the period from the second to 7th day after injury. The numbers of astrocytes increased and reached the highest level in the 3rd day after contusion while the reactive proliferation of astrocytes is only a minor phenomenon. PMID- 12536398 TI - [MBP content in serum after closed diffuse cranial injury in rats]. AB - The MBP content change in serum was observed by ELISA in rats with experimental closed diffuse cranial injury. The MBP content was 6.1633 +/- 1.5301 ng/ml (X +/- S) in the normal group, 11.3818 +/- 2.6574 ng/ml in the first group died immediately after injury, 10.8319 +/- 2.3135 ng/ml in the second group of 15 min after injury. This increased level of MBP was lasting in the following 3 days and returned to normal at the 4th and 5th day after injury. The difference of the MBP levels between the groups after injury and that in the normal group is significant (P < 0.01). PMID- 12536399 TI - [Forensic medical identification of sudden death due to tuberculosis of adrenal glands]. AB - Two cases died of tuberculosis of adrenal glands were reported. The pathological findings are presented and their medico-legal significance was also discussed. In particular, emphasis is placed on the importance of histopathologic examination of the adrenal glands in the tuberculous subjects dying suddenly and in whom no obvious cause of death was found at necropsy. PMID- 12536400 TI - [Forensic pathology studies of sudden death caused by pulmonary embolism]. AB - The analytic result of 50 sudden death cases caused by pulmonary embolism showed that their main causes were trauma (42%), heart diseases (32%), surgical operation (20%) and so on. In majority of these cases (80%), thrombus embolus came from deep veins of lower limbs, in some cases (20%) came from cardiac cavity. These cases occurred most frequently in the period of October to December and rarely in the period of April to June. Finally, some problems in forensic examination in this sort of sudden death were discussed in this article. PMID- 12536401 TI - [Immunohistochemical observation of actin in viral myocarditis]. AB - 10 specimens of heart in autopsy were immunohistochemically studied by using anti actin(At) antibody. In 5 cases with definite or suspected viral myocarditis according to Dallas criteria, actin in myocardium showed focal or diffuse weak staining, and occasionally showed focal absence of staining. In 2 cases with myocardial infarction, actin showed large pieces absent of staining, while actin in 3 cases with non-cardiac death showed strongly positive, well-distributed staining. The results indicated the immunohistochemical observation of actin in myocardium is useful for improving the sensitivity in diagnosing viral myocarditis and differentiating the slight injuries of myocytes caused by ischemia or inflammation. PMID- 12536402 TI - [Quantification of female teenagers' competence of sexual cognition]. AB - Compiling a questionnaire of sexual cognition for surveying 224 female teenagers (11 to 14 years old), the results showed that they were deficient in the knowledge of sexual physiology, mortality and law. And also, the deficiency had a high co-relationship with the age of surveyed female teenagers. It suggested that this questionnaire be valuable in the quantification of sexual cognition. PMID- 12536403 TI - [Allele and genotype frequency distribution of D19S400 locus in several populations]. AB - Using polymerase chain reaction and PAG electrophoresis, D19S400 Locus was investigated in the populations of Chinese, Germany, Slovakian, and American Black Population. 620 samples collected from the four populations were tested and a total of 11 alleles (*7 to *17) and 47 genotypes were found. The heterozygosity was observed between 0.78 and 0.88, and the discriminating power was between 0.9385 and 0.9664. The distributions of genotypes in the four populations were in accordance with Hardy-Weinberg equilibrium. Significant differences were observed in the distribution of allele frequencies among the three major racial populations (Mongoloid, Caucasian, Negroid). According to the results obtained in this study, D19S400 locus is a robust STR for forensic and genetic purposes. PMID- 12536404 TI - [Research on DNA typing in the forensic identification of formalin-fixed paraffin embedded tissues]. AB - Successful extraction of DNA from formalin-fixed Paraffin embedded tissues is important to DNA typing for forensic purpose. In the research, 26 samples were analyzed by method of PM and HLA-DQA1 loci typing. 24 of these samples were successful. It showed that this technique and method could be applied to individual identification and paternity test by use of these sorts of samples. PMID- 12536405 TI - [Polymorphic study of CSF1PO locus in Han population in Wuhan]. AB - Using the primers designed by Yoshida, the amplified fragment length polymorphism of the STR locus CSF1PO was studied. 8 alleles were determined from 312 unrelated individuals of Han population living in Wuhan. Sensitivity and suitability of the PCR-STR typing using Yoshida's primers were compared with the one using Hammond's. Results proved that Yoshida's method were more suitable in forensic identification than that of Hammond, especially in the typing of severely degraded DNA samples. PMID- 12536406 TI - [Polymorphism and haplotype study of Y-chromosomal multi-STRs loci in Han population in Guangzhou]. AB - Using polymerase chain reaction and silver stain, polymorphism and haplotype study of Y-chromosomal multi-STRs loci: DYS19, DYS389I/II, DYS390 were studied. 111 samples of male were collected from Guangzhou area. 5 alleles were determined in DYS19 locus, 4 alleles in DYS389I locus, 5 alleles in DYS389II locus and 5 alleles in DYS390 locus. Compared with other racial populations, differences of distribution of allele frequencies existed significantly. 72 haplotype were present in this study. PMID- 12536407 TI - [Application of solid-phase microextraction technique to the detection of amphetamines in urine by GC]. AB - A simple and rapid detection of nine amphetamines co-existing in urine was described. In the test, the method of solid-phase micro-extraction (SPME) by GC technique was used. Urine (1.0 ml), NaCl (0.3 g) and 4-phenylbutylamine (internal standard) were added into a vial (1.5 ml), then the sample was adjusted to pH 12 with 10% NaOH and sealed with a teflon-coated septum. After immersion of the SPME fiber (100 PDME) in the sample for 15 min, the SPME needle was inserted into the injection port of the GC and extruded for 3 min. The result showed that each peak from nine amphetamines compounds and internal standard was clearly separated. The calibration curves were linear from 0.2 to 15 micrograms/ml for most of five amphetamines with r between 0.9928-0.9995. The CV were less 10%. It is concluded that the method is simple, quick, accurate and useful for the practical detection of urine concentration of amphetamines. PMID- 12536408 TI - [Analysis of fluoroacetamide in biological samples with GC/NPD, GC/FID]. AB - A GC/NPD and GC/FID method has been developed for the analysis of fluoroacetamide (1081), using AC-20 and FFAP capillary column. The recovery is 85.1% (RSD 5.4%), 85.1% (RSD 7.1%) and 81.6% (RSD 10.1%) in liver, whole blood and stomach tissue, respectively. PMID- 12536409 TI - [Determination of metabolites of heroin in urine and discrimination of heroin abuse]. AB - This article describes a sensitive method that detects morphine, 6 monoacetylmorphine, morphine-3-glucuronide and codeine in urine for qualifying the abuse of heroin. The analytes were extracted by solid phase C18. The limits of detection (LOD) for morphine and codeine were 50 ng/ml and 50 ng/ml, respectively. The RSD of morphine and codeine were 11.3% (n = 5), and 14.2% (n = 5) respectively. For urine, it does not need to be hydrolyzed before extracted, and for all analytes, also need not to be derivated. The difference ratio of morphine and codeine in the chromatography can be used to discriminate between the abuse of heroin and the administration of compound liquorice mixture. PMID- 12536410 TI - [The effect of injuries on Fn synthesized by FBs and its implication in wound timing]. AB - cFn synthesized by FBs around the wound was detected by immunochemistry in vitro. Using the method of ELISA, The concentration of cFn in the culture media was observed. The concentration of cFn had no change within 30 minutes after injury, but started to increase after 1 hour of the injury and kept this trend within next five hours after injury. By static analysis, it can be obtained that the concentration of cFn in culture media is time-dependent after injury. PMID- 12536411 TI - [A study of HSP70 and NF in brain contusion timing]. AB - The changes of HSP70 and NF after brain contusion were studied by the immunohistochemistry method. HSP70 and NF were seen at 30 min after injury around the wound area, but faded away after 12-24 h. The result suggests that these two proteins could be used to estimate brain contusion time, to diagnose brain wound and to distinguish antemortem and postmortem brain contusion. PMID- 12536412 TI - [Analysis of the type, reason for death and cause of non-malpractice tangle]. AB - 206 autopsy cases related to the death of non-malpractice medical tangle from 1972 to 1998 were studied retrospectively. 111 (53.88%) were died of natural diseases, 82 (38.81%) inevitable complications and 13 (6.31%) unpredictable medical accidents. For the cause of medical tangles, the diagnosis of 92 cases (44.66%) was questioned. The other disputes focused on the infusion/blood transfusion (31 cases, 15.05%), the injection (20 cases, 9.71%), drug misuse and delay in making diagnosis or giving treatment (18 cases, 8.74% respectively). 13 cases (13.11%) involved complains on medical staff's rudeness and hospital management. PMID- 12536413 TI - [The changes of GFAP and PCNA after human brain contusion: an immunohistochemical study in forensic pathology]. AB - In order to find out changes of GFAP and PCNA after human brain trauma, samples of 38 cases were studied by the immunnohistochemistry method combined with quantitative image analysis. The gray degree and area of PCNA positive cells increased to the maximum at the 4th day and then decreased gradually. GFAP positive cells began to increase significantly 12 hours after contusion, reached the maximum at the 4th day and stayed in high level until the 7th day. These results suggest that: (1) GFAP, PCNA show regular changes with survival time after brain contusion; (2) GFAP and PCNA can be used as time markers of brain contusion from 2-20 days. PMID- 12536414 TI - [An experimental study on the effect of positional asphyxia on cardiac and pulmonary functions]. AB - The purpose of the study is to explore the effect of positional asphyxia on cardiac and pulmonary functions and its role in causing death. respiration, pulmonary artery pressure and cardiac output were measured by inserting the swan ganz into the pulmonary artery from the femoral vein. Results showed that: (1) the respiration was stable before hanging, getting faster during hanging, and becoming irregular at the terminal stage of hanging; (2) the pulmonary artery pressure increased significantly during and at the terminal stage of hanging as compared with that before hanging; (3) the cardiac output was stable before and during hanging, but decreased significantly at the terminal stage of hanging. It is concluded that hanging may cause respiratory imbalance and hypoxia, resulting in cardiac and pulmonary dysfunction. The death caused by hanging belongs to respiratory death, the characteristic of which is primary respiratory arrest followed by secondary cardiac arrest. PMID- 12536415 TI - [Expression of ET of early ischemic myocardium in rats]. AB - To study the expression of ET of early ischemic myocardium, we designed a model of early myocardial ischemia (EMI) in rats. The study was conducted with SABC method and HE staining. Cardiac myocytes of ischemia showed a little ET-positive at 60-min. Cardiac myocytes, endothelium and smooth muscle of vessel showed ET positive staining in the area of EMI after 120-min. Staining was negative for Ischemia myocardium at 30-min and for the control group. No change was seen with HE staining in all groups. The data indicates that ET-SABC method may reveal the injury of myocardial ischemia 60-min and longer and may provide a new marker to diagnose EMI in forensic pathology. PMID- 12536416 TI - [Automated fluorescent analysis of STR profiling and sex determination]. AB - Denaturing PAGE coupled with the ABI377 fluorescent automated DNA sequencer was used to test the performance and reproducibility of the automated DNA profiling systems at vWA31A, TH01, F13A01, FES, TPOX, CSF1PO and Amelogenin gene. The allele designation windows at the 7 genetic markers were established and implemented into the genotype reading software. Alleles differing in just 1 bp in length could easily be discriminated. Furthermore, the interpretation guidelines were outlined for the 7 genetic systems by investigating the relative peak areas of heterozygote peaks and relative stutter peak areas in various monoplex systems. Our results indicate that if the ratio between two peaks is equal to or higher than 0.404, a herozygote could be determined, otherwise the homozygote be made. PMID- 12536417 TI - [The MN typing of whole blood and bloodstains by the method of one-step sandwich ELISA]. AB - The samples of whole blood and bloodstain were MN-typed by the method of one-step sandwich ELISA, monoclonal antibodies include anti-M, N and glycern-protein. The lowest sample amount in this study is: for whole blood is 0.065 microliter, for bloodstain about 10 to 50 ng. 455 cases of fresh blood and 200 cases of fresh bloodstain are correctly typed. For 58 cases of old bloodstains, the success rate is 96.6%. PMID- 12536418 TI - [Detection of antidepressant and antipsychotic drugs in human hair and evaluation of the results]. AB - The presence of therapeutic drugs in the hair of psychiatric patients was investigated, and the method for simultaneous determination of antidepressant and antipsychotic drugs by GC/NPD and GC/MS was also established. Eight different psychotroic drugs were detected from the hair of 32 subjects, and chlorpromazine and clozapine concentrations in the hair were found to be dependent on the dosage and the correlation coefficients were 0.8047 (P < 0.001, n = 16) and 0.7097 (P < 0.001, n = 16) respectively. Segmental analysis demonstrated that there was a correlation between the subject's drug exposure history and the distribution of drug along the hair shaft. PMID- 12536419 TI - [Determination of lidocaine in human spinal cord by high-performance liquid chromatography]. AB - A RPHPLC method for determining lidocaine in human's spinal cord. Was established. The linear range of the method was 0.20-20.0 micrograms/ml(r = 0.9999). The detection limit was 0.02 ng/ml(S/N > or = 3) and the clean-up recovery of lidocaine in human spinal cord was 82.4%-92.7%. The CV% were no more than 6.0% (N = 4). No interfere by other compounds was observed in study results using this method. RPHPLC method shows an excellent recovery, high sensitivity and good precision. PMID- 12536420 TI - [Polymorphism of 13 STR loci for establishment of Chinese criminal DNA database]. AB - Allele frequency distributions for thirteen STR loci: D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D16S539, THO1, TPOX, CSF1PO, and D7S820, have been determined by multiplex amplification and subsequent automatic fluorescent detection. The result of statistical analysis shows that the 13 loci satisfy Hardy-Weinberg expectation. The data obtained from this study will be used as reference data for forensic DNA analysis in China. PMID- 12536421 TI - [Immunohistochemical studies on vascular endothelial growth factor (VEGF) for the postmortem diagnosis of sudden coronary death]. AB - The expression of VEGF in myocardium of the heart of sudden coronary death (SCD; n = 16) and of the control group (n = 15) was studied quantitatively by immunohistochemistry SABC method, image analysis and statistical process. The results showed that local infarcted myocardium of the heart from all SCD had strong positive expression of VEGF, especially in peri-infarction area. The rate of positive expression was 100%, while only two of the 15 in control group had mild positive expression sparsely in local myocardium. Image analysis and statistical process showed that the positive index in SCD group was 13.68 +/- 2.73, and in the control group 2.05 +/- 0.84. There was a significant difference between the two groups (P < 0.01). The results suggested that the immunohistochemical staining of VEGF with image analysis and statistical process be used as an objective marker for the postmortem diagnosis of SCD. PMID- 12536422 TI - [An experimental study on the relationship between early postmortem intervals and DNA content of spleen cells in rats by computerized image analysis]. AB - 15 rats were used in this experiment. After the rats were executed, spleen cells were collected and smeared in every hour within 24 hours, then fixed with formalin solution and stained with Feulgen's method. Mean and integral optical density and abnormal index of cell DNA were measured with a auto-TV-image system, and the data were statistically analyzed. Results showed that the DNA degeneration rate of rat spleen cells had a linear relationship with early postmortem intervals. PMID- 12536423 TI - [Expression of bcl-2 and Bax protein in the failing human heart--an inhibitor or accelerator of apoptosis?]. AB - In general, myocyte death in myocardial infarctions (MI) is attributed to necrosis, but recently the involvement of apoptosis has been suggested. Loss of myocytes is an important mechanism in the development of cardiac failure of either ischemic or nonischemic origin. The expression of two proto-oncogenes influence apoptosis. Myocytes with positive bcl-2 immunoreactivity were seen in the heart of acute MI, localized only in the salvaged areas surrounding the infarcted tissues. Myocytes with overexpressed positive Bax immunoreactivity were observed in the salvaged areas of old MI. PMID- 12536424 TI - [Validation studies on the analysis of the HLA-DQA1 locus using PCR and its application in forensic science]. AB - Validation Studies were conducted to evaluate the forensic applicability of HLA DQA1 locus. The locus was typed by a reverse dot blot approach and allele specific oligonucleotide probes were immobilized on a nylon membrane strip. Allele and genotype frequency for HLA-DQA1 locus was determined in shanghai Han population. The frequency data can be used in individual identification and paternity test. PMID- 12536425 TI - [Multiplex PCR fluorescent detection of DNA loci and its application in forensic science]. AB - A method is reported using PCR multiplex amplification and fluorescent-labeled primers to genotype D3S1358, D16S539, TH01, TPOX, CSF1PO, D7S820 and Amelogenin loci. It is simple, accurate, sensitive and very useful for personal identification and paternity testing. PMID- 12536426 TI - [DNA typing of old skeletons and teeth with fluorescence tagged short tandem repeats]. AB - In order to identify the individual status of old human skeletons and teeth, studies were conducted on DNA extraction. The extracted DNA was then amplified and automatic typed with 373A sequence's gene scan system (made by PE Company) and fluorescence tagged Hum vWA, Hum LPL kit. This is a rapid, convenient and effective method in DNA typing of old skeletons and teeth in forensic investigation. PMID- 12536427 TI - [The application of minisatellite MS31A MVR-PCR digital coding technique in forensic science]. AB - Using isotope incorporate amplification technique of special 31A, 31A-A, 31A-G primers and alpha-32PdCTP, the minisatellite MS31A (located at D7S21 loci) was studied. A rapid, simple, and accurate MVR-PCR technique was successfully established. The technique can be applied in individual identification minutes amples, such as blood stains, semen stains contaminated by vaginal fluid, hair and bones. The sensitivity analysis revealed that this technique could detect 1 ng genoma DNA. It is also discribed about the application of the method in 40 criminal cases of rape and murder. PMID- 12536428 TI - [Determination of fenfluramine in corpse using GC/NPD and GC/MS]. AB - A method was described for the determination of fenfluramine by GC/MS in blood, urine, liver, lung, kidney and heart tissues. After liquid-liquid extraction of fenfluramine in the specimens obtained from autopsy, the samples were qualitatively identified by GC/MS, and quantitatively determined by GC/NPD. 4 phenyl butylamine was used as internal standard. The concentrations of fenfluramine in blood, urine and liver were 7.8 micrograms/ml, 64.2 micrograms/ml and 31.3 micrograms/g respectively. The findings of autopsy and the results of toxicological analysis were also discussed. PMID- 12536429 TI - [Evaluation of competence for serve a sentence]. AB - In order to explore the factors influencing the evaluation of competence for serve a sentence, 102 cases of forensic psychiatric expertise were reviewed. A questionnaire of "scale of adaptability to punishment circumstance" was designed to investigate these cases. The results showed that the patients of psychotic mental disorders and the patients with history of mental disorders have a relatively high rate of non-competence of serving sentences, and that they can not adapt theirself to punishment circumstance. PMID- 12536430 TI - [Immunohistochemical study on myocardial acute ischemia reperfusion injury in rats with anti-FOS protein antibody]. AB - To investigate law of FOS protein induced by myocardial ischemia reperfusion (MI/R) in acute period, a model in 20 anaethetized SD rats was established. Rats with normal and ischemia were used as control groups. Specimens were studied immunohistochemically with c-fos antibody. After ischemia 20 minutes, followed by 30 minutes reperfusion, the area of MI/R showed nuclei of myocytes positive staining in cryosection slides. In C2 group, the area showed nuclei of myocytes (37.76% +/- 9.66%) positive staining weakly. In C3 group, nuclei of cardiac myocytes(40.34% +/- 3.32%) was significantly positive. In C4 group it began to attenuate(35.36% +/- 4.81%). The myocardium in normal and ischemic control groups showed negative staining. No changes were seen with HE staining. Our data indicated that immunohistochemical method may reveal acute MI/R injury of ischemia 20 min and reperfusion 30 min with anti-FOS protein staining and there is problely a peak between 60-120 min after reperfusion. It is possible that the method be used to diagnose sudden cardiac death in forensic medicine. PMID- 12536431 TI - [An experimental study of corpse decay degree and timing of death]. AB - In this study, we put dead pigs indoors, outdoors and underground respectively, and kept a detailed record of the natural decay process, especially the tension of the pig's abdomen produced by corruptive gas. The result showed that the decay speed was closely related to temperature, humidity, rainfall, position of corpses on the ground, insect parasitism. The obtained data could provide an important referential basis for inferring time after death. PMID- 12536432 TI - [Changes of potassium and sodium content in erythrocyte of rabbit cardiac blood after death and timing of death]. AB - We studied the changes of potassium and sodium contents in erythrocytes and the activity of erythrocyte membrane Na(+)-K(+)-ATPase in 40 cardiac blood samples of rabbit corpses at different postmortem intervals (PMI). The erythrocyte potassium content (RBCK) decreased linearly as the PMI increased (R = -0.829, P < 0.025). during postmortem 48 hours. The erythrocyte potassium content and the time after death had significant correlation. The activity of erythrocyte membrane Na(+) K(+)-ATPase had no significant changes over postmortem 48 hours. PMID- 12536433 TI - [HLA-DRB1 typing by PCR-sequence specific primers for paternity determination]. AB - HLA-DRB1 typing by PCR-SSP technique was first used for paternity testing. The analysis of 42 paternity cases shows that this identification method is simple, fast and reliable and has high exclusion probability of paternity (66.3%). This method can also be applied to the study of transplantation, HLA associated disease and anthropogenesis. PMID- 12536434 TI - [Segmental analysis of hair from meperidine abusers and evaluation of the results]. AB - Using meperidine abusers as subjects for this study, we detected in hair meperidine and its metabolites normeperidine, N-OCH3-meperidine and N-AC meperidine. The content of meperidine and normeperidine of 60 abusers were 103 +/ 130 ng/mg and 117 +/- 143 ng/mg respectively. Meperidine was stably present in hair for at least 20 months. Normeperidine level decreased from hair root to tip. The segmental analysis of hair revealed that there were correlations between the drug abuse history and the distribution of drug along the hair shaft, and between the dose of meperidine and its content presented in hair. PMID- 12536435 TI - [Determination of heroin metabolites in biological fluids, tissues and hair of heroin addicts using GC/MS-SIM]. AB - Using GC/MS-SIM, we examined morphine distribution in human bodies of 2 delayed death cases and analyzed content of morphine and 6-acetylmorphine (MAM) in hair specimens of 7 cases. After hydrolysis and extraction, MOR and 6-MAM were derived with acetic anhydride or BSTFA and then the derivatives were qualitatively and quantitatively analyzed by GC/MS-SIM. The results indicated that urine, bile and liver were the best samples to reveal the heroin-related deaths. Hair analysis has its unique advantage for its long time of abusing monitor, compared to biological fluids. PMID- 12536436 TI - [Studies on individual characters of juvenile delinquents in Fujian province]. AB - Ashenk's Character Questionnaire was adopted to study the relationship between juvenile delinquency and individual characters. 621 delinquents at the age of 13 19 and 1371 school students (as control group) were questioned, and results were statistically analyzed. For male delinquents of 16-19, scores of extroversion, psychosomatic and nervosity were significantly higher than for control group (P < 0.01). For male delinquents of 13-15, scores of psychosomatic and nervosity were significantly higher than for control group (P < 0.01). For female delinquents of 16-19, score of nervosity was lower than for control group (P < 0.05). Different types of delinquency had no statistical relations with characters. The study reveals that characters of psychosomatic and nervosity might be related with male delinquent crimes, and that stabilization with female delinquent crimes. PMID- 12536437 TI - [Interval of death analysis of 21 cases of cranio-cerebral injuries caused by accidental falling]. AB - Forensic pathology materials of 21 cases of cranio-cerebral injuries caused by accidental falling were statistically analyzed. The injury causes were discussed. The characteristics of the injuries were investigated and compared with that caused by falling from high, traffic accident and fighting. PMID- 12536438 TI - [Forensic medical examination of "5.8" Shenzhen air crash]. AB - The air crash of B-2928 on May 8 1997 in Shenzhen was reported, which caused 35 dead and 37 wounded. The interval and cause of death, degree of injuries, cause of the accident and individual identification were investigated. Forensic measures to deal with such tragedies were suggested and discussed. PMID- 12536439 TI - [Forensic pathology analysis of factors in causing trouble by schizophrenics]. AB - We analyzed the data of different schizophrenics. The data of those who caused trouble were compared to those who did not cause trouble and to data of other mental patients. It was found that schizophrenics had a higher rate to cause trouble and commit murder. Schizophrenics who made trouble got worse therapeutic efficacy. They seldom got married and were not supported by their family. It is suggest that keeping strict supervision and providing good treatment to schizophrenics be positive ways to secure our society. PMID- 12536440 TI - [Brainstem changes in death from head injury]. AB - The cortex and brainstem changes in death from head injury were observed. There were hemorrhage, edema and neuron necrosis in brain injury group, and also in disputed group of brainstem injury. PMID- 12536441 TI - [Forensic medical identification of death due to poisoning of tetrodotoxin in puffer fish]. AB - One case died of poisoning of tetrodotoxin in dry fugu fillet was reported. Through the detailed forensic examination for this lethal case, many questions about forensic medical identification of death due to poisoning of tetrodotoxin was discussed and its medico-legal significance was also discussed. In particular, emphasis was placed on the details of a case, extraction of TTX in specimen, quantitative detection of TTX, purplish mouth and nails, dilatation of stomach and intestines, excluding other causes and combining animal experiment in order to get definite forensic medical identification. PMID- 12536442 TI - [Cardic death and apoptosis of myocardium]. AB - TdT-mediated Dutp nick end labeling (TUNEL) method was applied to detect apoptosis of myocardium of corpses dying from cardiac death. For arteriosclerotic coronary heart death, diffused apoptosis was observed around the border zones of histologically infarcted myocardium, and in the epithelial and smooth muscle of arteries. Few apoptosis cells were present for hypertensive heart death, and none for corpses dying from falling and hanging. Myocardium apoptosis may provide a new target for heart failure. PMID- 12536443 TI - [Enzymehistochemical and immunohistological changes of skeletal muscle motor end plates and muscle fibers and their relation to the time of death]. AB - In order to investigate the relationship between the retrograde changes of the skeletal muscle and the time of death in various postmortem intervals (PMI), a systemic study of the enzymehistochemical activity of AChE, SDH, LDH, Ca(2+) ATPase and the immunohistochemical reaction of SYN in motor end-plates and muscle fibers was conducted in rats under different temperatures and at various PMI. The results were analyzed and compared by an image processing system. It was found that these changes were related to the PMI, especially AChE changes. The AChE could be used as a sign-enzyme of skeletal muscle to date death. PMID- 12536444 TI - [Evaluation of anti-muscle actin monoclonal antibody HHF35 in the diagnosis of early myocardial infarction]. AB - In order to explore the specificity of anti-muscle actin monoclonal antibody HHF35 in the postmortem diagnosis of early myocardial infarction, the changes of HHF35 staining in the infarcted myocardia and other non-infarcted myocardia with direct or indirect myocardial injury were studied with immunohistochemical S-P method. The results showed that the loss of HHF35 staining of different degrees was found in the infarcted myocardia, but also in the other non-infarcted myocardia with direct or indirect myocardial injury, such as cardiac contusion, myocarditis, hemorrhagic shock, electrocution, mechanical asphyxia. So it should be cautious in diagnosing early myocardial infarction with HHF35 immunohistochemistry. PMID- 12536445 TI - [PCR-based DNA analysis of vWA31A, TH01, F13A01, FESFPS, TPOX and CSF1PO]. AB - The allele and genotype frequencies of 6 tetranucleotide STR loci were investigated in a sample of 132 unrelated individuals from Chinese Han population. The PCR products were analyzed on 6% denaturing PAGE and detected using fluorescently labeled primers in an automated 377 sequencer(PE). All loci meet Hardy-Weinberg equilibrium. There was no random association of alleles among the 6 loci. The allele frequencies were compared with other population databases. Except locus vWA31A, the observed heterozygosity at other 5 loci was significantly lower than that reported in Caucasian and Black population studies. The calculated DP = 0.99999, PE = 0.9708, pM = 1.059 x 10(-5). The allelic frequency data can be used in forensic identification and paternity testing. PMID- 12536446 TI - [Population study of the CSF1PO, TPOX and TH01 STR polymorphisms in east China]. AB - Allele and genotype frequencies of four short tandem repeat loci were determined in a population sample from east China by using PCR and fluorescence techniques. After denaturing PAGE, 9 alleles were identified for CSF1PO, 8 alleles for TPOX, and 7 alleles for TH01. All loci met Hardy-Weinberg expectations. In addition, the theoretical values of these loci for paternity and forensic casework were evaluated. PMID- 12536447 TI - [Analysis of 50 cases of sexual dysfunction]. AB - Fifty cases of male sexual dysfunction were analyzed in order to investigate the incidence and causes of the erectile dysfunction in the different injuries. The results showed that the incidence of erectile dysfunction of the patients with brevic fracture and urethra injury is 62.50%, main caused by the damage of pudendal nervous and arterial system. The incidence in the patients with the spinal fracture and light spinal cord injury is 50%, caused by the damage of nervous system. The erection in the 17 cases of the patients with only one testis injury is normal. Two cases with bilateral testis injury have suffered from erectile dysfunction. PMID- 12536448 TI - [Medicolegal appraisals of 22 cases of ocular penetration]. AB - 22 cases of ocular penetration injury were analyzed and a clinical criteria of the injury's degree was proposed. The reasons causing ocular penetration and the characteristics of injuries were discussed. PMID- 12536449 TI - [The foundation of the injury model on cultured human fetal lung fibroblasts and its immunohistochemistry research]. AB - Cultured human fetal lung Fbs was first used to study the relationship between wound age and the change of cFn synthesized by Fbs in Vitro. A sterilized injector was applied to scrap the cultured confluent Fbs and a "wound" was obtained. Fbs around the wound was seen to change from a stationary stage to an active stage. Using the method of immunochemistry and image analysis system (IAS), cFn synthesized by Fbs around the wound was observed at once, 30 min, 1 h, 3 h and 6 h after injury. It was found that cFn began to change 1 h after injury and increased continually within 6 h. It suggests that the change of cFn synthesized by Fbs is time-dependent after injury and this approach breaks a new path in the study on estimation of wound age. PMID- 12536450 TI - [An experimental study on the relationship between the estimation of early postmortem interval and DNA content of liver cells in rats by image analysis]. AB - 15 rats were used in this experiment. After rats were killed, cells of liver were collected, smeared, fixed with Formalin in every hour within the first 24 hours then stained with Feulgen's method, With an auto-TV-image system, mean and integral optical density and abnormal index of cell DNA were measured and analyzed statistically. The results showed that the DNA degeneration rate of liver cells had a linear relationship to early postmortem period in rats. PMID- 12536451 TI - [The expression of splicing fibronectin variants of cutaneous injuries in rats]. AB - In situ hybridization with Digoxingenin labeled probes were used to build the relationship between incised cutaneous wounding intervals and the expression of splicing fibronectin variants (total, E III A and E III B Variants) in rats. The results showed that all forms of FN variants increased notably with the wounding time going on. E III A and E III B Variants were expressed abundantly 30 and 60 minutes after injury, while few in normal skin. PMID- 12536452 TI - [The incidence of medical tangles related to death--the first report of a retrospective study on 27 years forensic autopsy work of medical tangles]. AB - In order to investigate the characteristics of medical tangle autopsy, 318 autopsy cases of medical tangle from 1972-1998 were studied retrospectively. In the first report, the following conclusions were presented about the happening of medical tangles related to death: (1) The case number of medical tangle autopsy increased greatly from 23 in 1972-1983 to 295 in 1984-1998; (2) There was not much sexual difference in deaths; (3) There were two peaks in age distribution, 104 cases (37.21%) were below 10 years old, 161 cases (50.63%) were at the age of 20-49; (4) Most cases (195 cases, 61.32%) happened in middle size hospitals of cities and counties. In recent years, disputes in private-owned clinics were increasing; (5) The surgery (71 cases, 22.33%) and obstetrics (58 cases, 18.24%) were the most involved departments. The reasons resulting in medical tangles of 206 cases of non-malpractice and 99 cases of malpractice were discussed briefly. PMID- 12536453 TI - [Personal identification using multiplex PCR and fluorescent detection of STR and Amelogenin locus]. AB - Nine tetranucleotide short tandem repeat (STR) locus of D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820 and one Amelogenin loci were analyzed using multiplex PCR with fluorescent-labeled primers following capillary electrophoresis. This method is simple, accurate, sensitive and very useful for personal identification. PMID- 12536454 TI - [The evaluation of the forensic applications of D1S80, DQA1 + PM and CTT]. AB - Studies were conducted to evaluate the application of D1S80, PM + DQA1 and CTT to Forensic Science. The methods used to type D1S80, DQA1 + PM and CTT are simple, accurate, sensitive, and also were useful for personal identification and paternity testing. PMID- 12536455 TI - [Analysis of electrophoretic patterns of different body hairs of the same individual]. AB - Keratin contents of different hairs from the head, pubic, armpit and leg of one individual were analyzed by SDS-PAGGE. Samples were collected from 20 individuals. The results showed that the electrophoretic patterns of different body hairs of the same individual were almost identical. The relative contents of keratin composition from the same individual were tested by Laser Densimetic Scanning. No obvious difference was found. Electrophoretic keratin patterns and relative keratin contents of head hairs from different parts (crown, left side, right side, forehead and occiput) of the same individual were observed. No obvious difference was found. It is concluded the analysis of electrophoretic keratin pattern of human body hairs can be used as an important evidence for forensic individual identification. PMID- 12536456 TI - [Gas chromatography with dual nitrogen-phosphorus detection and flame ionization detection for screening abuse drugs]. AB - A dual-detection Gas chromatographic method for screening out abuse drugs is described. A single capillary separation column is connected to a nitrogen phosphorus detection (NPD) and a flame ionization detection (FID) by an Y connector. The ratio responded by NPD/FID is influenced by operational conditions. But the relative ratio is not influenced when carbetane is used as a reference. The ratio and the relative ratio can be used to screening abuse drugs. PMID- 12536457 TI - [An autopsy study of 7 sudden death cases due to acute hemorrhagic enteritis]. AB - 7 sudden death cases due to acute hemorrhagic enteritis were analyzed to investigate the age, sex, clinical symptoms, medical diagnosis and anatomic results. The results showed that, because the pathological changes were not typical and clinical symptoms were varid, It was easy to come to erroneous diagnosis and cause medical disputes. PMID- 12536458 TI - [Analysis on forensic medical appraisals and VEP characteristics of ocular trauma caused by traffic accidents]. AB - In order to investigate the characteristics of forensic medical appraisals of ocular trauma caused by traffic accidents, Visual Evoked Potential examinations were performed in 30 cases and the results were analyzed. It was found that, compared with 30 cases of ocular trauma caused by ordinary injuries, (1) the proportion of craniocerebral injuries (83%) and cranial nerve injuries (53%) was much higher, so were the proportion of diplopia (30%), optic never injuries (20%), and orbital wall fracture (20%). (2) because victims always claimed that their visual acuity was reduced by traffic accidents, it was difficult to identify simulated lower visual acuity by ordinary method, and VEP examination was often used. Different degrees of visual functional disorders showed different patterns in VEP, such as no VEP wave, lowed P1 wave amplitude, delayed P1 period, and so on. PMID- 12536460 TI - [Cystic fibrosis: current therapy]. PMID- 12536461 TI - [Cystic fibrosis: current therapy. Molecular genetic diagnosis]. PMID- 12536462 TI - [Cystic fibrosis: current therapy. Nutrition]. PMID- 12536464 TI - [Cystic fibrosis: current therapy. Diabetes]. PMID- 12536463 TI - [Cystic fibrosis: current therapy. Pancreatic enzyme supplementation]. PMID- 12536465 TI - [Cystic fibrosis: current therapy. Diagnosis and treatment of hepatobiliary diseases]. PMID- 12536466 TI - [Cystic fibrosis; current therapy. Infections]. PMID- 12536467 TI - [Cystic fibrosis: current therapy. Physical therapy and rehabilitation]. PMID- 12536468 TI - [Cystic fibrosis: current therapy. Fertility versus sterility]. PMID- 12536469 TI - [Cystic fibrosis: current therapy. Antibiotics]. PMID- 12536470 TI - [Cystic fibrosis: current therapy. Bronchodilators]. PMID- 12536471 TI - [Cystic fibrosis: current therapy. Inhalation therapy]. PMID- 12536472 TI - [Cystic fibrosis: current therapy. Inhalation antibiotics]. PMID- 12536473 TI - [Cystic fibrosis: current therapy. Topical (inhalation) steroids]. PMID- 12536474 TI - [Cystic fibrosis: current therapy. Inhalation therapy with recombinant DNAse]. PMID- 12536475 TI - [Cystic fibrosis: current therapy. Hemoptysis]. PMID- 12536476 TI - [Cystic fibrosis: current therapy. Pneumothorax]. PMID- 12536477 TI - [Cystic fibrosis: current therapy. Allergic bronchopulmonary aspergillosis (ABPA)]. PMID- 12536478 TI - [Mucoviscidosis: current therapy. Indications for oxygen therapy and non-invasive mechanical ventilation]. PMID- 12536479 TI - [Cystic fibrosis: current therapy. Indications for lung transplantation]. PMID- 12536480 TI - [Mucoviscidosis: current therapy. Gene therapy]. PMID- 12536481 TI - [Cloning and the basic characteristics of a novel cDNA induced by ischemia and reperfusion]. AB - OBJECTIVE: To identify and clone genes induced by ischemia and reperfusion. METHODS: cDNA fragments responding to ischemia and reperfusion in porcine myocardium were isolated by mRNA differential display and then the cDNA library from porcine heart was screened. After cloning, sequencing, and hybridization, the basic characteristics of the gene were analyzed. RESULTS: One cDNA fragment from a novel gene up-regulated by ischemia and reperfusion was identified and cloned subsequently; a novel full-length cDNA containing an open reading frame encoding 608 amino acid was obtained by screening the library. Sequencing results revealed that both DNA and amino acid sequences had no substantial homology to previously described DNA or protein sequences. Northern blot analysis confirmed that this gene expression in ischemia and reperfusion myocardium was significantly increased, and the expression was also able to be detected in normal porcine heart, liver, lung, kidney, spleen, intestine, brain, and skeletal muscle. Southern blot analysis clearly showed that this novel gene had high evolutional conservation in human beings, pigs, rabbits, and mice. CONCLUSION: The cloned gene is a novel one induced by ischemia and reperfusion, and it may play a very important role in the response to ischemia and reperfusion. PMID- 12536482 TI - [Contrast of losartan, fosinopril and amlodipine on cardiomyocyte apoptosis and left ventricular remolding in hypertensive rats]. AB - OBJECTIVE: To investigate effects of losartan, fosinopril and amlodipine on cardiomyocyte apoptosis, cardiac remolding in the spontaneously hypertensive rats (SHR). METHODS: SHRs were treated with lorsartan (SHR-L), fosinopril (SHR-F), amlodipine (SHR-A), and untreated (SHR-C) respectively for 8 and 16 weeks. Cardiomyocyte apoptotic index (APOI), left ventricular mass, left ventricular mass index (LVM, LVMI), and plasma and myocardium angiotensin II(PAng II, MAng II) concentrations were examined. RESULTS: 1. The systolic blood pressure was decreased similarly in all treatment groups in 8 and 16 weeks. LVMIs were reduced significantly in all treatment groups. LVMI was significantly lower in SHR-F group than that in other two treatment groups in 16 weeks. 2. APOIs were decreased significantly in SHR-F group in 8 weeks and in all treatment groups, especially in SHR-F group in 16 weeks. 3. Compared with SHR-C group in both periods, PAng II and MAng II were significantly increased in SHR-L group, but MAng II concentration was only decreased significantly in SHR-F group in 8 weeks, and in SHR-F and SHR-A groups in 16 weeks. CONCLUSION: Losartan, amlodipine, and especially fosinopril can inhibit cardiomyocyte apoptosis, prevent myocardial fibrosis, and reverse heart hypertrophy. Inhibition of myocardium rennin- angiotension--aldsteron system may be the mechanism of the three drugs' cardioprotective effects. PMID- 12536483 TI - [Influence of human cytomegalovirus infection on the expression of HOXB5, HOXB6, HOXB7, and HOXB8 genes in gliomaous cells]. AB - OBJECTIVE: The expressions of HOXB5, HOXB6, HOXB7, and HOXB8 genes of U251 cell infected by human cytomegalovirus and/or treated with all trans-retinoic acid(ATRA) were detected by semi-quantitative RT-PCR. The results were that U251 cell did not express HOXB5, HOXB6, and HOXB8 but expressed HOXB7 without infecting HCMV and/or being treated with ATRA. After infected with HCMV and/or treated with ATRA, the expressions of HOXB7 and HOXB8 were up-regulated and the expression of HOXB7 lasted the fourth generation, while HOXB8 was up-regulated only in the second generation. These results indicate that the abnormal expression of HOXB gene induced by HCMV may play an important role in the maldevelopment. PMID- 12536484 TI - [Analysis of synovial membrane apoptosis in different arthropathy]. AB - OBJECTIVE: To study the relationship between synovial membrane apoptosis and different arthropathy for the pathogenesis of cartilage lesion in arthropathy. METHODS: Synovial specimens (28 cases) were divided into control group (9 cases), osteoarthritis group (7 cases), dislocation group (5 cases), meniscus injury group (5 cases) and arthropyosis group (2 cases). Synovial histological sections were stained with methylene blue, and examined under microscope. DNA was extracted from synovial specimens, and analysed by gel electrophoresis. RESULTS: DNA ladder pattern was the most clear in the arthropyosis group, moderately clear in the dislocation group (especially in a case of 20 years-old shoulder dislocation), less clear in the osteoarthritis group, the least clear in the meniscus injury group, and was not seen in the control group. CONCLUSION: It can be concluded that the increased apoptosis is related to causes, course and seriously of arthropathy, and it might be a pathogenesis of cartilage lesion in arthropathy. PMID- 12536485 TI - [Effect of hyperinsulinism on NO production in vascular smooth muscle cells]. AB - Nitrate reductasing and immunoblotting test were used to investigate the modulation of protein kinase C (PKC) activity in cultured rat vascular smooth muscle cells (VSMCs) with high concentration of insulin (HI) and the effect of HI on lipopolysaccharides + gamma-interferon (LPS + gamma-IFN)-induced nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression with or without PKC inhibitor H7. The results were that membrane PKC activity preincubated with HI was significantly higher than that with the control group(P < 0.05), and NO production pretreated by HI was obviously lower than that of the control group(P < 0.01). PKC inhibitor H7 ameliorated the down-regulation of LPS + gamma-IFN induced NO production by high concentration of insulin. Immunoblotting test revealed a decrease induced by the high level insulin in the expression of iNOS in VSMCs. It is suggested that hyperinsulinism may activate PKC to partly inhibit the expression of iNOS and decrease NO production in VSMCs. PMID- 12536486 TI - [Changes of renal blood flow and serum NO2-/NO3- in rabbits treated with mannitol at different dosages]. AB - To observe the changes of renal blood flow and serum NO2-/NO3- in rabbits administrated with mannitol at different dosages, and investigate the mechanism of acute renal failure (ARF) induced by massive mannitol. Eighteen healthy male Zealand rabbits were randomly divided into 3 groups: minor mannitol group(n = 6, 8 g.kg-1 within 2 hours), control group (n = 6, saline of the same volume), massive mannitol group with free water taking(n = 6, mannitol 40-60 g.kg-1 with 3 days). The changes of renal function, renal blood flow and NO2-/NO3- in serum and urine were observed. The results showed that: no significant change in creatinine clearance (Ccr), renal blood flow (RBF) and serum NO2-/NO3- was found between minor mannitol group and control group. In massive mannitol group with free water taking, Ccr and RBF significantly decreased and serum NO2-/NO3- significantly increased (P > 0.05). The results suggest that ARF induced by massive mannitol is associated with a significant decline of RBF, on the other hand, endothelium derived relaxing factor(EDRF) maybe compensatively increase. PMID- 12536487 TI - [Treatment of cervical carcinoma in mice by injecting monoclonal antibody of epidermal growth factor receptor into solid tumor]. AB - OBJECTIVE: To observe the effect of treating cervical carcinoma in mice by injecting monoclonal antibody of epidermal growth factor receptor (EGFR-McAb) into solid tumor. METHODS: Six hundred and fifteen strain mice bearing U14 cervical cancer cell were subcutaneously injected with EGFR-McAb in Group A and B. The mice in control Group A and B were injected with 0.9% sodium chloride instead of EGFR-McAb. The volume of the tumor in Group A as measured for 2 weeks consecutively. The survival time of the mice both in Group A and in control Group A were observed. The mice in Group B were killed in the 4th, 7th, 10th, 14th, 16th, and 17th days. Tissue section of the tumor as stained immunohistochemically for CD31. Active areas of angiogenesis chosen under microscope and average microvessels counted per field(40x) were taken as intratumor microvessel density(IMD). RESULTS: (1) The mean survival time in Group A was 42.8 day while in control Group A it was 28.3 day. (2) The tumor growth inhibitory rate in Group A in the 4th, 7th, 10th, and 14th days was 13%, 42%, 83%, and 84%, respectively, and in the 7th, 10th and 14th days there was a significant difference(P < 0.05). (3) The Group B was 15.6 +/- 4.0, while in control Group B it was 31.1 +/- 14.6. The average IMD in the Group B was lower than that in the control Group B. CONCLUSIONS: An effective treatment to cervical cancer in mice can be made with EGFR-McAb. It is suggested that EGFR-McAb may be a target for treating cervical cancer. PMID- 12536488 TI - [Pharmacokinetics of high-dosage naftopidil capsules in dogs]. AB - OBJECTIVE: To establish a high-performance liquid chromatography (HPLC) for the determination of naftopidil in plasma, and make a study of pharmacokinetics of high-dose naftopidil capsules in dogs. METHODS: Five healthy dogs were treated with naftopidil capsules in an oral single dose of 200 mg. Naftopidil in dog plasma was determined by HPLC. The mobile phase consisted of acetonitrile and 0.05 mol.L-1 phosphate buffer (pH 6.5) (60:40), verapamil was selected as internal standard, and the eluate was monitored at 230 nm. Pharmacokinetics calculations were carried out by practice pharmacokinetics programs (Version 97, 3P97). RESULTS: The calibration curve of naftopidil was linear within the range of 10-1,200 ng.ml-1, and the correlation coefficient was 0.9992, the recovery rate of HPLC was (100.23 +/- 3.00)%, within day RSD and between day RSD were no more than 5.56% and 3.30% respectively. The plasma drug concentration-time course in dogs conformed to the one-compartment model. T1/2Ke value was (1.91-4.99) h, Tmax was (1.87-3.21) h, Cmax was (338.79-414.04) ng.ml-1. CONCLUSION: HPLC is accurate and reliable, and can be used for studying the pharmacokinetics of high dose naftopidil capsules in dogs. PMID- 12536489 TI - [Histomorphometric analysis in ovariectomized rat with osteoporosis]. AB - OBJECTIVE: To establish the pattern of bone loss of Sprague-Dawley rats after ovariectomy, to investigate and assess the pattern of bone remodeling in ovariectomized rats and the therapeutic action of nilestriol and levenorgestrel. METHODS: Histomorphometric analysis for bone tissues was performed with the instrument of computerized image analysis. RESULTS: The trabecular bone volume, mean trabecular thickness, and trabecular bone perimeter were all significantly decreased (P < 0.01) in ovariectomized rats in the 14th week postsurgery compared to the sham operated group. But the osteoblastic index, osteoclastic index, resorptive surface of trabecular, and trabecular surface covered by osteoblast were significantly increased (P < 0.01). There was no significant difference between the treatment group and the sham operated group. CONCLUSIONS: The pattern of bone loss of rats after ovariectomy has been established successfully. The study provides evidence that the cause of bone loss in the ovariectomized rats is attributable to the increase of bone turnover and negative imbalance in bone remodeling process. The treatment of nilestriol and levenorgestrel may decrease the bone turnover and correct the imbalance to cure the osteoporosis. PMID- 12536490 TI - [Analysis of the HLA A/B local haplotype among the females of Han nationality in Hunan province]. AB - OBJECTIVE: To investigate and analyse HLA A/B local haplotype gene frequency (HGF) in the females of Han nationality in Hunan. METHODS: HLA-A, B local antigen polymorphisms were investigated via microcytotoxicity assay. Haplotype was inferred by means of linkage disequilibrium parameter. RESULTS: Among the 46 haplotypes, HGF high were: A11-B60 (HGF = 0.1381), A2-B60 (HGF = 0.0861), A24-B48 (HGF = 0.0709). Three haplotype's linkage disequilibrium parameter have significant difference, they are A2-B52, A11-B13, A24-B52. CONCLUSION: The female of Han Nationality of Hunan HLA-A, B especially B local antigen frequency exists difference sompare with Han Nationality in Hunan and other areas in China of mix public of both sexes. When study relativity of the female disease of Han Nationality of Hunan with HAL, should notice HLA distributing particularity, be assure data veracity. Among the 46 haplotypes, A2-B52, A11-B13, A24-B52 three haplotypes exist significant linkage disequilibrium, probably they are own haplotypes at the female of Han Nationality in Hunan. PMID- 12536491 TI - [A case of primary clear cell carcinoma of lung]. PMID- 12536492 TI - [Relationship between the level of plasma D-dimer and diabetic microangiopathy]. AB - OBJECTIVE: To study the relationship between the level of plasma D-dimer and microangiopathy of the type 2 diabetes mellitus. METHODS: The level of plasma D dimer and blood viscosity were measured in 52 cases with type 2 diabetes mellitus, and these patients were divided into three groups according to the level of urinary albumin excretion (UAE) together with 15 age-matched healthy volunteers as the control (NC group). RESULTS: In the cases with higher urinary albumin excretion rate, including 28 cases with trace albuminuria (EDN group) and 10 cases with clinical albuminuria (CDN group), plasma D-dimer and blood viscosity were increases than 14 cases without albuminuria (SDM group) and NC group. D-dimer was positively correlated with diabetic retinopathy, Serum insulin and diabetic durations, but not correlated with blood viscosity and fibrinogen. CONCLUSION: The type 2 diabetes mellitus complicated with microangiopathy had a higher level of plasma D-dimer. As a marker of thrombus formation, D-dimer may play an important role in the pathogenesis of diabetic microangiopathy. PMID- 12536493 TI - [Efficacy of plasma exchange combined with fetal hepacyties on viral hepatitis gravis]. AB - OBJECTIVE: To evaluate the efficacy of mid-artificial liver support system (ALSS) on viral hepatitis gravis. METHODS: One hundred and thirty eight patients with hepatitis gravis were treated with plasma exchange combined with fetal hepacyties, fifty six patients were treated with plasma exchange and other forty eight patients were treated with fetal hepacyties respectively. The liver function was examined in all patients before ALSS. The liver function, amino acid spectrum and cardiac muscle enzyme were examined before and after ALSS in patients treated with plasma exchange and fetal hepacyties. RESULTS: It showed that the survival rate of the patients treated with plasma exchange combined with fetal hepacyties was higher than that of the patients only treated with plasma exchange or fetal hepacyties (P < 0.01). The liver function, BCAA/AAA ratio and cardiac muscle enzyme also significantly changed in patients treated with plasma exchange and fetal hepacyties before and after ALSS (p < 0.01 or 0.05). CONCLUSION: Plasma exchange combined with fetal hepacyties can effectively treat viral hepatits gravis. PMID- 12536494 TI - [Diagnostic value of transbronchial lung biopsy via fiberbronchoscope of peripheral or diffuse pulmonary disease]. AB - OBJECTIVE: To understand the diagnostic value of peripheral and diffuse pulmonary disease examined with Transbronchial lung biopsy (TBLB) via fiberoptic bronchoscope. METHODS: Four hundred and sixty patients of peripheral and diffuse pulmonary disease were diagnosed with TBLB via fiberoptic bronchoscope without fluoroscopic guidance. The result of clinical data and pathologic diagnosis were underwent retrospectively analyses. RESULTS: Confirmed diagnosis was obtained in 351 patients; the total positive diagnostic rate was 76.3%. Diseases were included lung cancer, pulmonary tuberculosis, diffuse interstitial pulmonary fibrosis, pulmonary sarcoidosis etc. The positive diagnostic rate of TBLB on pleural pulmonary disorders was the lowest. There were no any other serious complications except bleeding, blood sputum, fever and pneumothorax occasionally. CONCLUSIONS: TBLB in diagnoses of peripheral and diffuse pulmonary disease is a reliable, safe, handy cheap, may repeatable operational method. PMID- 12536495 TI - [Influence of low-molecular weight heparin on urine interleukin-6 in patients with proliferative glomerulonephritis]. AB - OBJECTIVE: To explore the influence of low-molecular weight heparin (LMWH) on interleukin-6 (IL-6) in patients with proliferative glomerulonephritis, 24 patients which had been distinctly diagnosed by renal biopsy were randomly divided into a control group (n = 12) and a treatment group (n = 12). In addition to the treatments of the control group, the patients of the treatment group were given with LMWH by hypodermic injection for four weeks. The level of urine IL-6 was detected by radioimmunoassay (RIA). The results were that the level of urine IL-6 was higher in patients than that in healthy persons (n = 24); after the treatment, the level of urine IL-6 was significantly decreased both in the control and treatment group, but it was lower in the treatment group than that in the control group. The effect of LMWH on patients with proliferative glomerulonephritis may be associated with inhibiting the proliferation of mesangial cell and the accumulation of mesangial matrix. PMID- 12536496 TI - [Effects of prostaglandin E1 on hepatic function and hepatofibrosis in patients with chronic hepatitis B]. AB - To search for an effective drug to cure patients with chronic hepatitis B. Twenty eight patients with chronic hepatitis B were treated with prostaglandin E1. The comprehensive indexes, including glutamic pyruric tranasaminase (GPT), serum total bilirubin (TBIL), total bile acid (TBA), hyaluronic acid (HA) and precollagen type III (PCIII), were examined before and after treatment. The levels of GPT, TBIL, TBA, HA, PCIII after 1 month of the treatment were significantly lower than those before the treatment (P < 0.05), there were significant differences between the treated and the controlled group patients with chronic hepatitis B (P < 0.05). The results suggest that the treatment with PGE1 might improve hepatic function, and resist the hepatic fibrosis in patients with chronic hepatitis B. PMID- 12536497 TI - [Clinical analysis of 8 cases of painful opthalmoplegia]. PMID- 12536498 TI - [Clinical evaluation of intermittent strophanthin K therapy for congestive heart failure combined coronary artery disease]. AB - OBJECTIVE: To evaluate the effects and safety of intermittent strophanthin K therapy (ISKT) for congestive heart failure combined coronary artery disease with sinus rhythm. METHODS: Two hundred patients divided into Group A (98 cases with maintenance digoxin therapy) and Group B (102 cases with ISKT). They were studied for 3 months and some of them for longer period. RESULTS: Comparing the pretreatment data: 1. heart rate (HR, bpm), 2. left ventricular ejection fraction (LVEF), 3. blood pressure (Bp, mmHg, calculated values as mean Bp for statistics). In group A, item 1. and 2. were significantly improved (item 1. 88 +/- 12 and 68 +/- 12, P < 0.01; item 2. 0.32 +/- 0.12 and 0.40 +/- 0.12, P < 0.01; item 3. showed no significant difference (126 +/- 21/90 +/- 6 and 128 +/- 21/80 +/- 5, P > 0.05). In group B, item 1., 2. and 3. were significantly improved (item 1. 90 +/- 10 and 70 +/- 11, item 2. 0.32 +/- 0.10 and 0.45 +/- 0.10, item 3. 128 +/- +/- 20/91 +/- 7 and 110 +/- 10/76 +/- 10, the p valves are the same < 0.01). As compared with the posttreatment data of both group A and B, HR, P > 0.05, there was no significant difference, LVEF, P < 0.05, there was significant difference, Bp, P < 0.01, there was significant difference. It showed no significant difference in total occurrence rate of digitalis overload or toxication between two groups also. CONCLUSION: ISKT for congestive heart failure combined coronary artery disease with sinus rhythm is effective and safe, with better improvement of heart function and Bp level. PMID- 12536499 TI - [Therapeutic effect of gamma-ray stereotactic radiotherapy on brain metastases]. AB - OBJECTIVE: To evaluate the therapeutic effects of gamma-ray stereotactic radiotherapy (SRT) plus whole brain irradiation and radiotherapy alone in brain metastases. METHODS: Forty-three patients with brain metastases were treated by SRT plus whole brain irradiation and 50 patients were treated by routine radiotherapy. SRT was given with the dosage of 15-27 Gy, and whole brain irradiation was given with the dosage of 30-40 Gy. RESULTS: One-year survival rate, median survival period, and tumor local control rate in SRT plus whole brain irradiation group were higher or longer than those in the routine radiotherapy group (P < 0.01). Mortality in SRT plus whole brain irradiation group was lower than that in the routine radiotherapy group (P < 0.05). CONCLUSION: The therapeutic effect of SRT plus whole brain irradiation on brain metastasis of cancer is superior to the routine radiotherapy. PMID- 12536500 TI - [Ultrasonographic evaluation of adrenal masses]. AB - OBJECTIVE: To identify the diagnostic value of ultrasonography on adrenal masses. METHOD: Retrospectively analyzed the ultrasonographic features of 116 patients who suffered from adrenal masses, which were verified histologically after operation. RESULTS: Of all patients, the correct rate of localization and determination is 94.8% and 93.6%, respectively. CONCLUSION: Ultrasonography can be considered as an accurate means of scanning adrenal masses. PMID- 12536502 TI - [Microendoscopic disectomy in the treatment of lumbar disc herniation]. AB - OBJECTIVE: To investigate the clinical value of microendoscopic disectomy for the treatment of lumbosacral disc herniation(LDH). METHODS: Microendoscopic disectomy (MED) technique was used to remove the nucleus and decompress the narrowed nerve root canal through an incision of 1.6 cm in length under the guidance of the "C" arm X-ray machine. RESULTS: Eighty-two patients with LDH underwent MED were followed up for 5 months. Eighty patients were evaluated as excellent, two as good according to the Macnab Scale. CONCLUSION: The microendoscopic disectomy can clearly expose the intervertibra and the nerve root, has little influence on the stability of the posterior structure of the spine, and can keep the spine intact. So the microendoscopic disectomy is a safe and an effective technique. PMID- 12536501 TI - [PLA2 activity in serum and cutaneous tissues in patients with psoriasis vulgaris]. AB - We examined the activity of PLA2 by quick simple trace titration method in the serum and cutaneous tissues in patients with psoriasis vulgaris patients than that in the control group. The activity of PLA2 was significantly higher in psoriasis vulgaris patients than that in the control group. PLA2 activity in sera of psoriasis patients in the active stage or rash area > 30% was higher than that in the inactive stage or rash area < 30%. It is suggested that the sera PLA2 activity may be related to the disease activity in patients with psoriasis vulgaris, and PLA2 may play an important role in the pathogenesis of the psoriasis. PMID- 12536503 TI - [Detection of brain vascular diseases with CT angiography]. AB - OBJECTIVE: To identify the diagnostic effect of spiral CT in brain vascular diseases. METHODS: Sixty four patients underwent CT angiography, including 22 patients with subarachnoid hemorrhage (SAH), 6 patients with oculomotor nerve paralysis, 5 patients with brain substance hemorrhage, 6 patients with arteriovenous malformation (AVM), 4 patients with mass or aneurysm, 5 patients with brain infarction, 16 patients with epilepsy or headache and dizzy. 8 cases also underwent digital subtraction angiography (DSA), 4 cases were performed operation in 64 cases. RESULTS: Spiral computed tomography angiography (SCTA) was all succeeded in 64 cases. 26 aneurysms and 8 AVMs were found by SCTA; in 6 of 8 cases of AVMs, their supply arteries and drainage veins were clearly displayed; SCTAs of 5 cases with substance hemorrhage were normal; in 1 of 5 cases with cerebral hemisphere infarction, its internal carotid obstruction was demonstrated by SCTA; the results of 12 SCTAs were identical with the results of their DSA or operation. CONCLUSION: SCTA is a useful non-invasive inspection method for detecting brain vascular diseases such as aneurysms and AVMs. PMID- 12536504 TI - [Associated sedation of propofol and midazolam in small dosage and gastroscopy]. AB - OBJECTIVE: To search for a sedative method that is more suitable for gastroscopy. METHODS: All of patients were randomly divided into control group and experimental group. The experimental group was treated with propofol and midazolam, the control group was treated with propofol alone. The cumulation dosage of propofol, sedative effect, variation of BP and SaO2 were observed in all patient. RESULTS: The cumulative dosage of propofol in the experimental group was lower than that in the control group [(73.21 +/- 18.67) mg and (117.23 +/- 21.57) mg respectively]; the oblivious degree in the experimental group was higher than that in the control group (95.65% and 80.00%); the onset time and the descendant range of BP and SaO2 were also lower in the experimental than those in the control group. There was not remarkable difference in sedative effect and veriviscont time between the control group and the experimental group. CONCLUSIONS: In such a rapid operation of gastroscopy, the dosage of propofol in the experimental group is obviously less than that in the control group, while it does not affect the effect of sedation, the diagnose and cure time in gastroscopy room, and has more security and less cost. PMID- 12536505 TI - [Relationship between basic fibroblast growth factor (bFGF) and cell proliferation, invasion in pituitary adenoma]. AB - Basic fibroblast growth factor (bFGF) and proliferating cell nuclear antigen (PCNA) were examined by immunohistochemistry in fifty-one pituitary adenomas. The results showed that the intensity of bFGF expression was strong or intermediate in twenty-nine adenomas, which was positively related to cell proliferation, there was positive relationship between bFGF expressin and tumor invasion in non functional adenomas. The data suggest that bFGF may play some role in the origin and progression of pituitary adenomas. PMID- 12536506 TI - [Relationship between phenomenon of acquired activated protein C resistance and antiphospholipid antibodies in patients with systemic lupus erythematosus]. AB - OBJECTIVE: To determine the occurrence of activated protein C resistance (APCR), to identify APCR is associated with thrombotic events (TEs), and acquired APCR is associated with the presence of antiphospholipid antibodies (APLAs) in 30 patients with systemic lupus erythematosus (SLE). METHODS: Laboratory tests included dilute Russell's viper venom time assay for LA (dRVVT-LA), ELISA assay for ACL, APC sensitivity ratio, and factor V Leiden were detected by PCR-Mnl/I digestion. RESULTS: Acquired APCR was presented in 14(46.67%) of 30 patients. Factor V Leiden was not found in any patients. The incidence of TEs in the APCR positive patients was significantly higher than that in the APCR-negative patients (42.85% vs 6.25%, P < 0.05). The incidence of TEs in the LA-positive patients was also significantly higher than that in the LA-negative patients (50% vs 11.1%, P < 0.05). CONCLUSIONS: The presence of either APCR or LAs is associated with one of the risk factors of TEs (P < 0.05). There is not a significant interaction between APCR and LAs in the association with TEs. Acquired APCR may not reflect the interference of LAs with the protein C pathway which may represent a mechanism of LA-associated TEs. PMID- 12536507 TI - [Study of self-threading pins in the preservative restoration of teeth with large area defects]. AB - OBJECTIVE: To investigate the application of self-threading pins in the preservative of teeth with large area defects. METHODS: A follow-up study of 42 out of 50 patients with large area defects in vital teeth were restored by self threading pins. RESULTS: Among the cases that had been followed up for 5 years, except 2 upper molars which were extracted because of fraction, other cases achieved good effects. CONCLUSION: Self-threading pins are very suitable to restore large area defects of teeth with healthy pulp, and can be widely used because of significant clinical effects. PMID- 12536508 TI - [Insulin and hyperosmotic glucose solution external used for treating pressure sore]. AB - OBJECTIVE: To observe the efficacy of insulin and hyperosmotic glucose solution for treating pressure sore. METHODS: Using moist burn ointment as the control drug, comparisons were made between the two groups in terms of average healing time. RESULTS: In trial group the average healing time of II degree pressure sore was 11.6 +/- 2.7 days, while in the control group 12.9 +/- 3.4 days, The difference was not statistically significant. The average healing time of III degree pressure sore in trial group was significantly shorter than that of the control group (22.3 +/- 4.3 days vs 24.8 +/- 3.9 days, P < 0.05). CONCLUSION: Insulin and hyperosmotic glucose solution is effective in treating pressure sore. PMID- 12536509 TI - [Changes in plasma von Willebrand factor and nitric oxide levels in patients with pregnancy induced hypertension]. AB - OBJECTIVE: To study the changes and clinical significance of plasma von Willebrand factor (vWF) and nitric oxide (NO) levels in patients with pregnancy induced hypertension (PIH). METHODS: Plasma vWF and NO were detected by ELISA and Griess assay respectively in 36 patients with PIH and 18 healthy pregnant women. RESULTS: The levels of plasma vWF in patients with moderate and severe PIH increased significantly. than those in healthy pregnancy. The levels of plasma NO in patients with PIH were significantly lower than those in healthy pregnancy. The higher the vWF levels or the lower the NO levels, the more severe in PIH. There was a negative correlation between the plasma vWF and NO levels. CONCLUSION: The results suggest that plasma vWF and NO concentration can be used as indicators in the judgement of the severity of PIH. PMID- 12536510 TI - [Early operation with anterior screw-plate system for cervical spine fracture and dislocation]. AB - OBJECTIVE: To evaluate the function of the cervical spine anterior screw-plate system in the early operation of cervical spine fracture and dislocation. METHODS: One hundred and fifteen patients with fracture and dislocation of lower cervical spine treated by cervical anterior decompression, reduction, iliac crest autograft, fixed with cervical anterior locking plate (AO, spine-tech, Orion). RESULTS: All the cases were free from complication except one patient was complicated with infection of the surgical wound and another one with a transient dysfunction of recurrent laryngeal nerve. Symptom of patients were significantly improved in postoperation and no hardware failure. CONCLUSIONS: Early anterior decompression operation for cervical fracture and dislocation can achieve satisfactory clinical outcomes, anterior cervical vertebral screw-plate system is safety, simplicity, rigidity and can provide good conditions for early anterior cervical spine operation. PMID- 12536511 TI - [Application of suboccipital craniotomy by transconfluence sinuses in treatment of riding transverse sinuses hematoma]. AB - OBJECTIVE: To study the best methods of operation for riding transverse sinuses hematoma. METHODS: Analysed 12 patients with riding transverse sinuses hematoma treated by transconfluence sinuses suboccipital craniotomy. RESULTS: Of twelve cases, 10 cases were cured, 1 cases was in severe deformity and the rest 1 died after operation. CONCLUSION: The advantages of suboccipital craniotomy are anatomical replacement, less postoperative complications caused by suboccipital craniotomy, so suboccipital craniotomy by transconfluence sinuses is a better operative way in treating riding transverse sinuses hematoma. PMID- 12536512 TI - [Simultaneous determination of catecholamine levels in plasma by high performance liquid chromatography]. AB - OBJECTIVE: To use a high performance liquid chromatography with electrochemical detection to simultaneously determine catecholamine(CA) levels in plasma and to diagnose chromaffin cell tumors and neuroblastoma. METHODS: The plasma samples after flowing extraction by ion-moderated partition were determined with electrochemical method to detect CA levels in plasma; the lowest detective limit, the precision, recovery, sensitivity of CA levels were tested and a reference range was established based on the respective data of 18 healthy persons. RESULTS: The recovery rates of epinephrine and norepinephrine were 86.2% and 90.5%. The method of testing epinephrine was linear at the range of 0.2 to 10.6 nmol.L-1 with 0.05 nmol.L-1 of the lowest detective limit. The intra- and inter assay coefficients of variation were lower than 10.7% and 11.1%, respectively. CONCLUSION: The method is simple, accurate, sensible in the clinical diagnosis of CA levels. PMID- 12536513 TI - [Study of HCMV infection in newborns by PCR and ELISA]. PMID- 12536514 TI - [Clinical application of bronchography with spiral CT scan]. PMID- 12536515 TI - [Clinical analysis of 20 patients with Crohn's disease]. PMID- 12536516 TI - [Measurement of serum IgG subclass concentration in systemic lupus erythematosus]. PMID- 12536517 TI - [A case report of cerebra resuscitation with subhypothermia therapy after penetrating cardiac trauma]. PMID- 12536518 TI - [Alterations of acid-base and electrolyte in the pigs of piggyback orthotopic liver transplantation]. PMID- 12536519 TI - [Study on the fusion expression of FBXO30: a novel member of F-box protein family]. AB - OBJECTIVE: The cDNA sequence of the coding region of FBXO30 (F-box only protein 30), which is a novel member of F-box protein family, was cloned into the mammalian expression vector pEGFP-C2 by non-directional cloning method and introduced into the NIH 3T3 cells by liposome transfection. Observation under the fluorescent microscopy after transfection showed that EGFP (enhanced green fluorescent protein)/FBXO30 was expressed and existed mainly in the cytoplasm. At the same time, the cDNA sequence of the coding region of FBXO30 was cloned into prokaryotic expression vector pGEX-4T-2 by directional cloning strategy. The GST (glutathione S-transferase)/FBXO30 fusion protein was expressed under the induction of IPTG (isopropylthio-beta-D-galactoside) in E. coli. A new band (approximately 65.5 kD) was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. Purification of the GST-fused FBXO30 was carried out by affinity chromatography with glutathione sepharose 4B. The fusion expression of FBXO30 indicates that: FBXO30 protein is a cytoplasmic soluble protein, and the stable fusion expression of FBXO30 in the prokaryotic expression system can provide a base for the preparation of the specific antibody of FBXO30 and further functional study of FBXO30. PMID- 12536520 TI - [Screening of mouse connexin 31 genomic clone and mapping by fluorescence in situ hybridization]. AB - Connexin 31(Cx31) is one of the human genetic deafness disease genes. To determine the structure and function of Cx31 gene of mouse, the mouse Cx31 gene clone was screened from the genomic library with the probe obtained according to the sequence of the mouse Cx31 gene cDNA. Cx31 gene was mapped to the center of mouse chromosome 4 by fluorescence in situ hybridization. The present study lays a foundation for researches on structure and function of Cx31 gene, as well as establishment of the relative transgenic mice and gene knockout mice. PMID- 12536521 TI - [Retrospective analysis of 102 cycles of intracytoplasmic single sperm injection in treating severe male infertility and fertilization failure]. AB - The aim of this study is to retrospectively analyze 102 cycles which received intracytoplasmic single sperm injection (ICSI) in our lab from January 1998 to December 2000. With the improvements of the technique and environment of the microscopic manipulation from February 2000, the survival rate (90.6%). fertilization rate (73.3%) and embryo development rate (97.3%) of the later stage were significantly higher than those of the previous (68.8%, 39.0%, 78.7% respectively). The clinical pregnancy rate reached a higher level (41.3% and 21.4% respectively). The ICSI technique established currently in our lab can effectively treat the severe male infertility and fertilization failure. PMID- 12536522 TI - [9 cases of heterotopic pancreas]. PMID- 12536523 TI - [Ischemic preconditioning inhibits ischemia/reperfusion induced myocardial apoptosis in rabbits in vivo]. AB - OBJECTIVE: To investigate the effects of ischemic preconditioning on myocardial apoptosis induced by ischemia/reperfusion. METHODS: Twenty four rabbits were randomly allocated to three groups(n = 8), pseudo-operation group(group P), ischemia/reperfusion group(group IR), and ischemic preconditioning group (group IP). Group IR and group IP were subjected to three hours of left anterior descending coronary artery occlusion followed by three hours of reperfusion. Ischemic preconditioning was achieved by three 5-minute cycles of ischemia, each followed by 5 minutes of reperfusion. Infarct size and area at risk were defined by dual staining with triphenyltetrazolium chloride and Evans blue dye. DNA laddering in the border zone myocardium of ischemic area at risk was revealed with agarose gel electrophoresis, and apoptosis index(AI) was obtained with flow cytometry. RESULTS: Infarct size, expressed as a percentage of the area at risk(IS:AR), was(60.8 +/- 10.8)% in group IR. Ischemic preconditioning reduced myocardial infarct size significantly to (33.1 +/- 4.9)% (P < 0.05). AI was (13.83 +/- 3.98)% in group IR, and were reduced significantly to (5.85 +/- 1.59)% in group IP(P < 0.01). DNA laddering indicative of fragmented DNA was clearly demonstrated in myocardial specimens sampled from the lateral border zones of the ischemic area at risk in group IR but were attenuated in group IP. CONCLUSION: It suggests that ischemic preconditioning inhibits the rabbit myocardial apoptosis induced by ischemia/reperfusion in vivo. PMID- 12536524 TI - [In vitro culture of modified neonatal pig islets]. AB - A cell-isolation-and-cell-culture-in-vitro method which can produce a mass of new born pig islet-like clusters was introduced. The new-born pig pancreas were incubated in V-type collagenase (0.5-1.0 mg.ml-1) for 8-12 minutes, and then cultured in vitro for 7 days before large amount of highly purified porcine islets were produced. The neonatal pig islets had good insulin secretion and normal sub-cell structure. The results suggest that the neonatal pig islets are probably a fine donor source for clinical xenotransplantation of islets. PMID- 12536525 TI - [Stable expression of human cytochrome P450 2E1 cDNA in nasopharyngeal cancer cell line CNE-2]. AB - OBJECTIVE: This study as designed to investigate the function of CYP2E1 protein in chemical carcinogen metabolism in vitro and its action mechanisms in nasopharyngeal carcinogenesis. METHODS: DAN recombination technique was used to construct a eukaryotic expression vector pcDNA3.1-2E1 of human cytochrome P450 2E1 cDNA. The vector was introduced into a nasopharyngeal cancer cell line CNE-2 mediating by lipofectamine. After these transforming cells were treated with G418 for 2 weeks, several G418-resistant clones were selected and amplified in culture. RESULTS: The authors acquired two cell clones CNE2-2E1-1 and CNE2-2E1-2 with stable integration and expression of exogenous CYPE2E1 cDNA. After Southern blot analysis, RT-PCR and Western blot were fulfilled. CONCLUSION: The cell models setting here will be a good tool in the further study of CYP2E1. At the same time, its use will help to screen chemical carcinogenesis and to elucidate the mechanism of carcinogenesis which is related to chemical carcinogen activated by CYP2E1. PMID- 12536526 TI - [Growth behavior and metastatic pattern of cervical cancer U14 transfected with human papillomavirus in inbred mouse]. AB - OBJECTIVE: To investigate the growth behavior and metastatic pattern of murine cervix cancer U14 transfected with human papillomavirus(HPV) in inbred 615-strain mouse. METHODS: We transfected HPV 16 E6 and E7 genes into mouse cervix carcinoma cell strain NO. 14(U14) by electroporation and liposome, respectively. The transfectants were selected by G418, and several high-expressed HPV16 E6 and E7 clonal cell lines (E6+ U14, E7+ U14) were detected by PCR and by immunohistochemistry. We transplanted those cells into inbred 615-strain mice both subcutaneously and intraperitoneally to observe the growth behavior and metastasis of them. RESULTS: The durations of tumor appearance were 5-7 d, 11-14 d, and 8-10 d after having been transplanted subcutaneously with wild type U14, E6+ U14, and E7+ U14, respectively(P < 0.05). The mean survival times of mice were 29 d, 43 d, and 35 d, respectively. Metastasis could be found both in lymph nodes (90%, 30%, and 40%, respectively) and lungs (60%, 10%, and 20%, respectively). After intraperitoneal inoculation, the mean survival durations of mice were 14.2 d, 20.6 d and 18.3 d. We could not find metastasis both in lymph nodes and lungs. CONCLUSION: Murine cervix cancer U14 cells transfected with HPV16 E6 and E7 have different growth behavior and metastatic patterns after transplanted in inbred mice, which provide useful models for studying their immunotherapy or other strategies for cervical cancer with E6 and E7 as a target. PMID- 12536527 TI - [Operative correction of pectus accompany with atrial septal defect]. PMID- 12536528 TI - [Protective effects of scopolamine on rabbits with acute brain injury]. AB - OBJECTIVE: To study the protective effects of scopolamine (Sco) (a muscarinic receptor antagonist) on rabbits with acute brain injury, and preliminarily explore the roles of acetylcholine (ACh) in the early pathologic changes of traumatic brain injury(TBI). METHODS: The model of acute brain injury was established by a free-falling device, and the rabbits received Sco intraperitoneally at 5 minutes and 2 hours after brain injury respectively. Transcranial Doppler (TCD) was used to monitor cerebral blood flow velocity (CBV) and pulsatility index (PI) of middle cerebral artery (MCA). The levels of Ca2+, the activities of superoxide dismutase (SOD), the contents of malondiadehyde (MDA) and Evans blue (EB) were measured, and the pathological changes were observed in brain tissue. RESULTS: This model consisted with the pathologic changes of accelerated brain injury. Sco reduced the levels of Ca2+, EB and MDA, increased the activities of SOD, and improved cerebral blood flow following brain injury. CONCLUSIONS: In the early stage of brain injury, ACh can cause neuron calcium overload, oxide free radical reaction, cerebral vasospasm, and increase blood-brain barry(BBB) permeability. Scopolamine, a muscarinic receptor antagonist, can improve these pathologic changes. As a result, Sco has protective effects on brain tissue injury. PMID- 12536529 TI - [Kindling model of SD rats induced by injection of PTX intraperitoneally]. AB - OBJECTIVE: To observe the kindling model of Sprague-Dawley (SD) rats induced by injection of different doses of picrotoxin (PTX) intraperitoneally, and to evaluate the effect of the drug on the model. METHODS: SD rats were devided randomly into four groups: control group, epilepsy Group I (PTX, 1.5 mg.kg-1.d 1), epilepsy Group II (PTX, 2 mg.kg-1.d-1) and the phenobarbital group. The kindling model of rats was induced by daily injection of PTX intraperitoneally. The seizures in the rats were observed, and the results of electroencephalography (EEG) were recorded. RESULTS: On the 20th day after injection of PTX, 71% SD rats were kindled in epilepsy Group I; 4 rats were dead, 2 rats were at stage IV of convulsions, and 2 rats were at stage V of convulsions in epilepsy Group II; 17% rats were kindled in the phenobarbital group. The EEG demonstrated spike discharges in the kindled rats. CONCLUSIONS: In this models, the better dose of PTX is 1.5 mg.kg-1.d-1, and phenobarbital sodium can protect SD rats injected PTX intraperitoneally from seizure. PMID- 12536530 TI - [p16 and cyclinD1 protein expression and p16 gene mutation in primary human hepatocellular carcinoma]. AB - OBJECTIVE: To elucidate the relationship between expression of p16 and cyclinD1 proteins and evaluate the role of p16 gene exon 2 mutation in hepatocellular carcinogenesis. METHODS: Streptavidin-peroxidase conjugated method(SP) was used to detect expressions of p16 and cyclinD1 proteins in 44 cases of hepatocellular carcinoma(HCC), and polymerase chain reaction single-strand conformation polymorphism analysis(PCR-SSCP) to detect p16 gene mutation in exon 2 in 12 cases of HCC and liver tissues adjacent to HCC. RESULTS: Expression rate and positive signal intensity of p16 protein in HCC were significantly lower(P < 0.01) and those of cyclinD1 protein in HCC were significantly higher (P < 0.05) than those in pericarcinomatous tissues. Of 12 fresh HCC tissues, p16 gene mutation in exon 2 was found in 2 cases, whereas that was not found in pericarcinomatous tissues. CONCLUSIONS: 1. Inactivation or/ and deletion of p16 protein may be one of important reasons which result in proliferation unbalance of cells. 2. p16 gene mutation in exon 2 presents in HCC, but it does not frequently occur in Chinese hepatocarcinogenesis. 3. p16 gene abnormality and cyclinD1 over expression may coact in HCC. PMID- 12536531 TI - [Inflammatory-inducing effect of lumbar disc tissue: an experimental study]. AB - OBJECTIVE: To study whether herniated or degenerative lumbar disc tissues have the potentials to produce or induce peripheral blood white cells to produce inflammatory mediator phospholipase A2 (PLA2), and to find the effective inhibitors of PLA2. METHODS: Ten specimens of lumbar discs were obtained when patients were performed operations. The peripheral blood samples were drawn out from the same ten patients (7 ml per patient). Mononuclear white cells were separated from each peripheral blood sample. Then, cell culture was performed according to the following combinations: Group 1 was mononuclearcytes; Group 2 was disc tissue; Group 3 was devitalized disc tissue + mononuclearcytes; Group 4 was devitalized disc + mononuclearcytes + dexamethasone; Group 5 was devitalized disc + mononuclearcytes + artemisinin. The activity of PLA2 was detected at last. RESULTS: Compared with Group 3, other groups had a significant difference, (P < 0.01). Group 5 compared with Group 4 had no significant difference (P > 0.05). CONCLUSIONS: 1. Some components in disc tissue can induce mononuclearcytes to produce high activity of PLA2 which is an inflammatory mediator very possible to cause back pain and radiculopathy in lumbar disc herniation. 2. Artemisinin has similar effectiveness to dexamethasone to inhibit the production of PLA2 in vitro. PMID- 12536532 TI - [The effects of bizhongxiao decotion (BZX) on T-lymphocyte subsets in the peripheral blood of patients with rheumatoid arthritis]. AB - OBJECTIVE: To observe the effect of BZX on T-lymphocyte subsets in the peripheral blood of patients with rheumatoid arthritis (RA) and study the mechanism of curative effects of BZX in immunization. METHODS: 42 patients with RA in active stage were divided into 1. treatment group with BZX 2. control group with western medicine and twenty healthy control group (HCG) was arranged before treatment. Their CD4+ and CD8+ were tested and the ratio of CD4+ cells to CD8+ cells was calculated before treatment and after 3 months treatment. RESULTS: In the peripheral blood of patients with RA, CD4+ was more than that of the HCG. but CD8+ was lower and the ratio of CD4+ cell to CD8+ cells was higher than that of the HCG. After 3 months treatment, CD4+ decreased, CD8+ increased and the ratio of CD4+ cells to CD8+ cells decreased. In addition, all results were similar to the normal. We found there were no significant differences between the two treatment groups in the results after treatment. CONCLUSION: There were cell immunoregulation disorders in patients with RA. BZX can regulate the disorder of T-lymphocyte subsets, help rebuild the immune steading and have effects of immune suppression similar to MTX combined medicine. PMID- 12536534 TI - [Location of the pre-excitation part in Wolff-Parkinson-White syndrome by Doppler tissue imaging]. AB - OBJECTIVE: To evaluate the feasibility of newly developed Doppler tissue imaging (DTI) for locating the early contraction sites in Wolff-Parkinson-White (WPW) syndrome. METHODS: The time-sequential changes in ventricular wall motion of thirty-three patients suffering from WPW syndrome were examined by DTI. The pre excitation site was represented as a red region appearing on the myocardium right after the delta wave of the electrocardiogram and confirmed by electrophysiology. RESULT: Of 21 patients with left-sided accessory pathways, the early contractive sites were correctly identified by DTI in 18 patients, while the sites were determined by DTI in 8 of 12 patients with right-sided accessory pathways. There was no significant difference between the two groups (P > 0.05). CONCLUSION: The DTI system is helpful to localize the accessory pathways and evaluate the results after radiofrequency ablation. PMID- 12536533 TI - [Experimental study of bovine bone morphogenetic protein combined with sintered bone in the treatment of bone defects]. AB - OBJECTIVE: To confirm the capability of ectopic bone induction of the bovine bone morphogenetic protein (bBMP), and investigate the bBMP combined with sintered bone (SB) in the treatment of bone defects. METHODS: bBMP was separated from the bovine cortical bone and implanted into the muscle pouches of the balb/c mouses. 4 mouses were killed at the 1st, 2nd, 3rd and 4th week, and their tissues were fixed in 4% formalin for histologic examination. The bBMP was combined with sintered bovine cancellons bone (SB), and segmental defects were made at raddi in 16 New Zealand rabbits. One side of the rabbits was implanted with bBMP-SB, and another side implanted with SB. The animals were killed in the 2nd, 4th, 6th and 8th week after operation for X-ray and histologic examination. RESULTS: One week after bBMP implantation, the chondrocyte was induced in muscle pouches, the woven bone was seen in 2 weeks, and the trabecular bone was formed in 4 weeks. In bone defects models, the X-ray and histologic examination showed that all parameters (chondrocyte induction, the mount of the trabecular bone, defects heal) were better in the animals treated with bBMP-SB than that with SB. CONCLUSION: 1. bBMP can induce ectopic bone formation. 2. bBMP-SB can improve the healing of the bone defects. PMID- 12536535 TI - [Expression of the neural cell adhesion molecule in human astrocytomas]. AB - OBJECTIVE: To investigate the expression of the neural cell adhesion molecule (NCAM) in human astrocytomas, and evaluate the relationship between NCAM and proliferating cell nuclear antigen (PCNA). METHODS: Forty-eight astrocytomas were studied by SP method with monoclonal antibody against NCAM and PCNA proteins. RESULTS: All normal brain tissues expressed NCAM; NCAM expression was positive in Grade I-II glioma [95% (19/20)], Grade III glioma [38.9% (7/18)], Grade IV glioma [20% (2/10)] respectively. An inverse correlation was observed between the degree of NCAM expression and that of PCNA (r = -0.657, P < 0.05). CONCLUSION: NCAM is a down-regulated factor in the development of the malignancy of astrocytomas; and it is suggested that the reduced NCAM expression might be involved in the development of biological malignancy. PMID- 12536536 TI - [Detection of serum type IV collagen and the alteration of this collagen in various liver diseases]. AB - OBJECTIVE: To detect serum Type IV collagen levels in patients with various hepatic diseases. METHOD: A sandwich enzyme-immunoassay, using two monoclonal antibodies to recognize 7S and TH of Type IV collage, was used to detect serum levels of Type IV collagen in 428 patients with various liver diseases. RESULTS: Serum levels of Type IV collagen in healthy control subjects were (63.2 +/- 31.9) microgram.L-1. Type IV collagen levels in the serum of patients with heavy hepatitis were the highest [(801.8 +/- 431.2) microgram.L-1]. All Types of hepatic diseases were significantly higher than those in healthy subjects (All P < 0.01). CONCLUSION: The assay used in this study is simple and practical and can be applied to clinical evaluation of hepatic fibrosis. PMID- 12536537 TI - [Detection of telomerase activity level in human non-small-cell lung cancer]. AB - OBJECTIVE: To investigate the telomerase activity in human non-small-cell lung cancer (NSCLC) and to investigate the possibility of telomerase as a tumor biological marker. METHODS: The semi-quantitative assay and expression of PCR based telomeric repeats amplification Protocol ELISA (TRAP-PCR-ELISA) and PAGE (TRAP-PCR-PAGE) were detected in 15 NSCLC tissues and 15 tumor-adjacent tissues, 3 normal lung tissues. RESULTS: No telomerase activity in 15 tumor-adjacent tissues and 3 normal lung tissues was detected, However, of 15 NSCLC tissues, 13 were positive for telomerase activity with a 86.7% positive rate, telomerase activity was significantly higher in NSCLC than in Tumor-adjacent tissues and normal lung tissues (P < 0.001). CONCLUSION: The results suggest that telomerase is a tumor specific gene marker. PMID- 12536538 TI - [Efficacy and safety of low dose amiodarone for paroxysmal atrial fibrillation in the aged patients with no-valvular heart diseases]. AB - OBJECTIVE: To study the efficacy and safety of low-dose amiodarone (AD) for paroxysmal atrial fibrillation in the aged patients with no-valvular heart diseases. METHOD: 40 inpatients were treated with load-dose AD (600 mg.d-1) for 6 days and then low maintenance dose of 50 mg or 100 mg daily. Clinical efficacy was evaluated with 24 h dynamic electrocardiogram. RESULTS: AD maintained efficaciously sinus rhythm about 90.0% (36/40) during 6 months, about 83.3% (25/30) during 12 months, about 72.7% (16/22) during 24 months. Only 2 patients were stopped oral AD because sinus bradycardia(< or = 45 bpm) occurred. CONCLUSION: Low dose AD (50-100 mg.d-1) is a safe and efficient maintenance of sinus rhythm in aged patients with no-valvular heart diseases and paroxysmal atrial fibrillation. PMID- 12536539 TI - [Analysis on three-year follow-up results of excimer laser photorefractive keratectomy in treatment for myopia and myopic astigmatism]. AB - OBJECTIVE: To evaluate the efficacy of excimer laser photorefractive keratectomy(PRK) on myopia and myopia astigmatism. METHODS: PRK was performed with VISXC20/20 on 377 eyes of 209 patients. Treated eyes were divided into two groups, according to their conditions before operation: Group I included 238 eyes (-1.50 to -6.00 D); Group II, 139 eyes (-6.25 to -16.00 D). All patients were followed-up for more than three years. RESULTS: Percentages of uncorrected visual acuity (UCVA) > or = 10/20 and 20/20 were 99.6% and 85.3% in Group I, and 79.9% and 48.2% in Group II respectively. Percentage of diopter < +/- 1.00 in Group I was 94.5%, and 61.9% in Group II. The rate of corneal haze of grade 0 was 100% in Group I, and 98.6% in Group II, but corneal haze of grade 2 was found in 1.2% of eyes in Group II. The postoperative intraocular pressure (IOP) of all operated eyes was normal. CONCLUSION: The results suggest that excimer laser PRK is an effective method for treating myopia and myopic astigmatism, especially for low and moderate myopia. PMID- 12536540 TI - [Prevention and treatment of the complications for whole-scale cataract surgery in rural hospitals]. AB - OBJECTIVE: To investigate the method of prevention and treatment of the complications of cataract surgery carried out on a large scale concentrated effort in rural hospitals. METHOD: The cataract extraction combined with intraocular lens (IOL) implantation in 394 eyes of 385 cases were analyzed in review. RESULTS: In 394 eyes, IOL implanting rate was 86%, cornea edema was observed in 8 eyes, hemorrhage anterior chamber in 2 eyes, secondary glaucoma in 1 eye, and IOL ccentricity in 3 eyes. The corrected visual acuity of these patients after being discharged from hospitals was distinctly improved comparing with pre-operation (P < 0.01), the rate of visual acuity relieved from blindness was 98%, and the rate of visual acuity relieved from visual handicap was 86%. CONCLUSION: The attentive examination before operation, careful manipulation at the time of operating and conscientious observation of treatment after operation are the keys of preventing and decreasing post-operation complications. PMID- 12536541 TI - [Relationship between serum C-reactive protein and creatine kinase isoenzyme in acute myocardial infarction]. AB - OBJECTIVE: To investigate the relationship between serum c-reactive protein(CRP) and creatine kinase isoenzyme (CK-MB) in the diagnosis of acute myocardial infarction (AMI). METHODS: We assessed the serum CK-MB and CRP concentration in 52 patients with definite AMI at different time after their infarction. RESULTS: 12 hours after infarction there were 100% and 44.2% patients with positive results in CK-MB and CRP respectively, and 48 hours are 57.7% and 86.5%. By continuous observation we found that the peaks of CK-MB and CRP occurred respectively at 12 hours and 48 hours after infarction. CONCLUSION: It is suggested that as a diagnostic marker CRP is superior to CK-MB in sabacent myocardial infarction patients. PMID- 12536542 TI - [Clinical research on the relationship between the orthondontic treatment and the temporomandibular joint noises]. AB - The sign of the temporomandibular joint(TMJ) noise was assessed through the comparison among seventy orthodontic patients before and after orthondontic treatment. The results showed, after orthondontic treatment the TMJ noise was less than before, no significant correlation was found between TMJ noise and the orthondontic treatment with or without tooth extraction. PMID- 12536543 TI - [A case of Marfan syndrome in midtrimester of pregnancy]. PMID- 12536544 TI - [Sequential BiPAP following invasive mechanical ventilation in COPD patients with hypercapnic respiratory failure]. AB - OBJECTIVE: To estimate the feasibility and the efficacy of extubation and sequential bilevel positive airway pressure (BiPAP) in chronic obstructive pulmonary disease (COPD) patients with exacerbated hypercapnic respiratory failure. METHODS: All patients were randomly divided into a control group and an experimental group. In addition to conventional therapy, the experimental group was treated with bilevel positive airway pressure via facial mask immediately, the control group was treated with conventional therapy alone. RESULTS: There were no difference in clinical characteristics and gas exchange at entry. pH value, PaCO2 and PaCO2 were significantly improved in experimental group following the first 45 minutes ventilation (P < 0.05), but no additional improvement was observed when ventilation was extended to 12 hours (P > 0.05). Reintubation rate was higher in the control group compared with the experimental group (P < 0.05). CONCLUSION: BiPAP can be well tolerated, and can improve ventilation function markedly. It can also facifiated the extubation, and reduced the reintubation rate in regular mechanical ventilation. PMID- 12536545 TI - [Analysis of 136 children with gastrointestinal hemorrhage]. AB - OBJECTIVE: To investigate the pathogeny of children with gastrointestinal hemorrhage, pathogeny distribution and the cause of misdiagnosis before gastrointestinal endoscopy check-up. METHODS: We detected 136 children suspected upper gastrointestinal hemorrhage (UGH) by gastrointestinal endoscopy and among them a few cases were diagnosed by celiotomy. RESULTS: One hundred and three cases of UGH, 7 cases of small intestine bleeding, and 4 cases of gulping blood syndrome were diagnosed; but the pathogeny of the other 22 cases was not clear. CONCLUSION: Gastric ulcer and duodenal ulcer are the common causes of gastrointestinal hemorrhage of elder children and inheriting alimentary canal abnormality leads to gastrointestinal hemorrhage of infants. Some children with bellyache were misdiagnosed as ascariasis or enterospasm. The endoscopy examination should carry on within 48 hours after the latest bleeding to reach the highest positive rate. PMID- 12536546 TI - [Treatment of complicated hypospadias in children]. AB - OBJECTIVE: To study the operation results of complicated hypospadias. METHODS: 29 cases of complicated hypospadias in recent 5 years were studied retrospectively. All cases were operated for the second time, methods used were urethroplasty with pedicled prepuce or transmitted penis prepuce tube, urethroplasty with penis scrotum skin tube in-situ and urethroplasty with combination of scrotum skin tube and transmitted prepuce tube. RESULTS: Except 2 fistulae sequela occurred after urethroplasty with penis skin tube in-situ in 1 case, all the other 28 cases were a success, the success rate was 96.5%. CONCLUSIONS: The key point for success is precise appraisal before operation and appropriate choice of operation method. PMID- 12536547 TI - [Changes of serum soluble tumor necrosis factor receptor I in patients with rheumatic fever]. AB - OBJECTIVE: To study the changes and the effects of soluble tumor necrosis factor receptor I (sT-NFR I) in patients with rheumatic fever, and to find out the laboratorial marker of clinic activity in patients with rheumatic fever. METHODS: The serum concentrations of sTNFR I in 16 patients with ARF, 16 patients with chronic inactive rheumatic heart disease (CRHD) and 15 healthy controls were measured by methods of ELISA and the relationship between sTNFR I and ESR or CRP were studied. RESULTS: The sTNFR I serum concentrations in patients with ARF (3.92 +/- 1.28) ng.ml-1 were significantly higher than those with CRHD (1.43 +/- 0.66) ng.ml-1 and healthy controls (1.06 +/- 0.42) ng.ml-1 (all P < 0.01). The serum concentrations of sTNFR I in patients with CRHD were almost the same with those in healthy controls (P > 0.05); Their positive correlation was significantly observed between sTNFR I and CRP(P < 0.001), and wasn't observed between sTNFR I and ESR. CONCLUSION: The Patients with ARF have high serum levels of sTNFR I. It may be used as the laboratorial marker in the clinic assessment of activity in patients with rheumatic fever. PMID- 12536548 TI - [Establishment of rabbit model for peritoneal dialysis related peritonits]. AB - OBJECTIVE: To establish a rabbit model corresponding with clinical characteristics of peritoneal dialysis related peritonitis. METHODS: Twelve New Zealand rabbits were randomly divided into peritonitis group and control group. They were infused standard living Staphylococcus aureus 3 x 10(9) colony forming unit (cfu) (peritonitis group) or 0.9% NaCl 1 ml (control group). After 48 hours, Laboratory rabbits in acute uremic model underwent regular dialysis for 6 hours. At different points of time, the concentration of glucose in dialysate and the dialysate/plasma (D/P) ratio of the concentration creatinine (Cr), total protein(TP), album(Alb) and IgG, the counts of leukocyte in dialysate and water ultrafiltrate were determined. RESULTS: D/D0 were significantly lower in the peritonitis group than that in the control group (P < 0.05). The counts of leukocyte and D/P ratio of Cr, TP, Alb and IgG were significantly increased in the peritonitis group (all P < 0.05). The drained volume was lower in the peritonitis group than that in the control group (but P > 0.05). CONCLUSION: In the rabbit model infused intraperoneally staphylococcus aureus, (1) transport peritoneum of low and large molecular weight solutes is increased, (2) the volume of ultrafiltrate is decreased, (3) the counts of leukocyte in dialysate is increased. These change in this model is basic all correspond with the clinical feature of patients of peritoneal dialysis related peritonitis. So this rabbit model of peritoneal dialysis related peritonitis is a successful experimental infection model. PMID- 12536549 TI - [Determination of 17-hydroxycorticosteroid by capillary zone electrophoresis]. AB - OBJECTIVE: To detect and dissociate 17-hydroxycorticosteroids(17-OHCS) in urine by using high performance capillary zone electrophoresis(CZE) so as to solve the problem that phenolic compounds interfere with the determination of urine 17-OHCS by chromatographic-spectrophotometry. METHODS: Pigments were absorbed from samples by activated kaolin in which the internal standard was theophyline. Urine 17-OHCS was absorbed by neutral styrene-divinylbenzene resin. Thus pigments and phenolic compounds were separated roughly. Then 17-OHCS was washed down by alcohol and separated and detected by capillary zone electrophoresis. RESULTS: Urinary 17-OHCS was separated successfully. Urinary 17-OHCS had a good linearity in the range of 2-50 mg.L-1 (r = 0.994). The average recovery rate was 96.17%. The relative standard deviation was less than 5%. The consistence detection limit was 0.3 mg.L-1. CONCLUSIONS: Compared with chromatographic-spectrophotometry, the high performance capillary electrophoresis has much less interference and is more simple, rapid, and accurate. PMID- 12536550 TI - [Application of the fiber bronchus lens in difficult endotracheal intubation]. PMID- 12536551 TI - [DNA sequence analysis of a novel variate type of PML/RAR alpha fusion gene transcript]. PMID- 12536552 TI - [Effects of angiotension II receptor antagonist on the level ET and NO in the patients with renal hypertension and protection of renal function]. PMID- 12536553 TI - [Cloning, sequencing of beagle E-selectin genomic DNA and comparison of it with other species]. AB - OBJECTIVE: Selectins are a group of members of Cell Adhesion Molecular family; they play very important roles in inflammation and neovascularization. The aim of this study is to clone, sequence Beagle E-selectin genomic DNA and make homogeneous analyses to compare it with those of other animals. METHODS: Twelve fragments of genomic DNA of Beagle E-selectin were obtained by PCR, and the 12 fragments were cloned in T-vector. At last, the genomic DNA sequences of Beagle E selectin were obtained through gene sequencing, assembling and base calling, and the gene structure was analyzed. RESULTS: The sequence of Beagle E-selectin genomic DNA that contained the whole open reading box was acgured. The results of homogeneous analyses showed a great extent of E-selectin gene homology among human, porcine and dog. CONCLUSION: The demonstration of a large extent of the gene homology has substantiated in this connection the important roles of E selectin in transplantation immunity and neoplasm development. PMID- 12536554 TI - [The blood drainage of Langerhans islets with special reference to the functional and clinical significance of pancreatic exocrine part]. AB - OBJECTIVE: This study sought to gain an insight into the blood drainage of Langerhans islets with special reference to the functional and clinical significance of pancreatic exocrine part. METHODS: Observations were carried out in 11 human dead bodies, 40 monkeys, 24 dogs, 62 rats and 24 rabbits by scanning electron microscopy and light microscopy with the technique of retaining microcirculatory dynamic and tissular information in the static specimens, microvascular serial reconstruction, and/or by intravital fluorecence microscopy with FITC-labeled erythrocytes. RESULTS: Three patterns of islet drainage channels drained into different acinar regions with different features and insulo insular drainage channels observed in the primates were found in this study. The authors suggest that these patterns might be termed as follows: 1. Continuous drainage vessels. All islets possessed these capillary-sized portal vessels which ran a short distance, and then drained into the peri-islet acinar region. 2. Convergent drainage vessels. Some islets possessed one or two, occasionally more, of these portal vessels, which were relatively long and/or thick and drained into the acinar region far away from the islet in the lobule. 3. Translobular drainage vessels. These portal vessels crossed the interlobular septum into an adjacent lobule where sometimes no islet existed, and then drained into the exocrine acinar region 4. Insulo-insular drainage vessels. Some islets in the monkey and human, possessed these vessels which drained into an adjacent small islet through the insulo-insular drainage vessels. CONCLUSION: Langerhans islets possessed the consummate drainage system which drained into different exocrine acinar regions, suggesting that the release of islet hormones is in some way necessary for the exocrine secreting function, and that the reduction of local insular hormone levels in insuloacinar portal circulation and the impairment of insular drainage vessels would be the morphological basis of the pancreatic exocrine pathologic lesion in human diabetes. PMID- 12536555 TI - [Activation of NF-kappa B induced by stretching and the gene expression of beta MHC regulated by NF-kappa B in cardiac myocytes]. AB - OBJECTIVE: To determine the mechanism of reexpression of beta-MHC gene caused by the stimulation of stretch. METHODS: The specimens were taken from the 1-, 2- and 3-day old SD rats. Immunohistochemical method was applied to observe the activation of NF-kappa B in stretched cardiac myocytes cultured on the silicon membrane, and then with the system of image analysis, the beta-MHC protein content was detected in four groups where NF-kappa B was activated or not activated, respectively. RESULTS: NF-kappa B was activated significantly by stretching cardiac myocytes for five hours. The beta-MHC protein content in stretched myocytes increased significantly, compared with that in the control myocytes, NAC-treated myocytes, and stretch-treated myocytes in the presence of NAC. CONCLUSION: The above data demonstrate that NF-kappa B signaling system is involved in regulating the stretch-induced gene expression of beta-MHC. PMID- 12536556 TI - [The role of YIGSR and RGD peptides in regulating fenestrae of sinusoidal endothelial cells]. AB - OBJECTIVE: To investigate the effects of Tye-Ile-Gly-Ser-Arg (YIGSR) and Arg-Gly Asp (RGD) polypeptides on the fenestrae of the liver sinusoidal endothelial cells (SECs). METHODS: By in situ collagenase perfusion and two-step percoll gradient centrifugation, SECs were isolated from normal Wistar rats and cultured on collagen type I or laminin-coated coverslips. The population of SECs fenestrae and their diameters were observed using scanning electron microscopical technique; and the synthesis of collagen type IV in SECs was assessed by radio immunoassay (RIA). RESULTS: The average number of fenestrae and their diameter in the SECs cultured on laminin-coated coverslips were significantly lower than those on collagen type I-coated coverslips. After the treatment with YIGSR pentapeptides (50 micrograms/ml) and RGD tripeptides (50 micrograms/ml) together for 48 hrs, the number and diameters of fenestrae in the SECs that grew on laminin-coated coverslips increased markedly. Meanwhile, the YIGSR and RGD peptides could significantly decrease the synthesis of collagen type IV in SECs that grew on laminin-coated coverslips. CONCLUSION: The results suggested that the YIGSR and RGD polypeptides may have an important role in regulating the morphology and function of SECs. PMID- 12536557 TI - [Molecular cloning and expression in E. coli of the surface-exposed lipoprotein LipL41 gene of Leptospira lai]. AB - OBJECTIVE: To construct a recombinant expressive plasmid using LipL41 gene of leptospira lai for further research of subunit vaccine of leptospira. METHODS: A pair of oligonucleotide primers were designed based on the sequence of LipL41 of leptospira kirschneri RM 52 in Genbank. With genomic DNA of leptospira lai 017 as template, a fragment was amplified by PCR and DNA sequencing analysis showed this fragment to be the gene that encodes LipL41. A recombinant vector was constructed using plasmid pGEX1 lambda T and the expression of LipL41 gene was tested. RESULTS: The production of PCR was LipL41 gene. The recombinant plasmid was constructed and testified by nuclease digestion and PCR, and LipL41 gene could express in E. coli. CONCLUSION: The recombinant plasmid has been constructed successfully and LipL41 gene can express stably at high level. PMID- 12536558 TI - [Pyruvate oxidase gene from Streptococcus sanguis: molecular cloning and sequence analysis of the gene]. AB - OBJECTIVE: To clone and sequence the gene of pyruvate oxidase (Sopox) from Streptococcus sanguis. METHODS: The PCR primers for Sopox gene were designed and synthesized according to the sequence of pyruvate oxidase (spxB) gene of S. pneumonia. The amplified PCR product was cloned into pUC18 and then subcloned into M13mp18 and M13mp19. The DNA sequence of the gene was analyzed. RESULTS: Sopox gene was successfully amplified from S. sanguis ATCC10557. The nucleotide sequence of the whole gene was revealed to be 1788 base pairs with one open reading frame coding pyruvate oxidase with 591 amino acid residuals. CONCLUSION: The clone and DNA sequence of Sopox gene were obtained which could serve as a foundation on which to elucidate the molecular mechanisms of hydrogen peroxide production and its regulation by oral streptococci. PMID- 12536559 TI - [Effect of para-aminobenzoic acid on the adhesion of Streptococcus mutans]. AB - OBJECTIVE: To investigate the effects of Para-aminobenzoic acid (PABA) on the adhesion of Streptococcus mutans to saliva-coated hydroxyapatite (S-HA). METHODS: The adhesion model in vitro established by Gibbons was adopted in this study to quantify adsorptive cells through [3H] thymidine labelling. RESULTS: PABA solution over a range of concentration from 10(-9) to 10(-3) g/L could inhibit the attachment of S. mutans to S-HA effectively, and the inhibition declined when the concentration of PABA solution was 10(-3) g/L. CONCLUSION: PABA can inhibit the adherence of Streptococcus mutans to S-HA effectively. PMID- 12536560 TI - [The expression of BDNF and NT-3 in dorsal root ganglion following peripheral and central axotomy--an immunohistochemical study]. AB - OBJECTIVE: To explore the expression of brain-derived neurotrophic factor (BDNF) and neurotrophin-3(NT-3) in dorsal root ganglion (DRG) following peripheral and central axotomy. METHODS: Fifteen male adult cats were divided into intact group, peripheral axotomy group and central axotomy group. The ten cats of the latter two groups were subjected to unilateral peripheral cut and central processes cut respectively. The animals were allowed to survive 4 days long; L5 DRG was taken from each animal and made into frozen sections 20 microns thick. The sections were stained using antisera of BDNF and NT-3 respectively by immunohistchemical ABC method. The neuronal numbers of BDNF and NT-3 of the three groups were counted and compared. RESULTS: The positive products of BDNF were mainly distributed in small to medium sized neurons (13-57 microns) while those of NT-3 were mainly in large sized neurons (57-100 microns). The neuronal numbers of BDNF and NT-3 in peripheral and central axotomy groups (24.1 +/- 3.5, 29.4 +/- 5.2; 19.4 +/- 2.4, 20.2 +/- 1.9) increased significantly than those of intact group (12.1 +/- 2.2, 13.3 +/- 2.9), P < 0.01. And the neuronal numbers of BDNF and NT-3 in peripheral axotomy groups were more than those of central axotomy group(P < 0.05). CONCLUSION: The results indicate that peripheral axotomy and central axotomy lead to the increase of the expression of BDNF and NT-3 in DRG neurons. And the effect of peripheral axotomy is greater than that of central axotomy. PMID- 12536561 TI - [A study of on the transport of neurotrophin-4 in facial nerve]. AB - OBJECTIVE: This is a study on the transport information of neurotrophin-4 (NT-4) in facial nerve using radioactive tracer technique. METHODS: After one lateral facial nerve trunk of adult rabbit was transected, a silicone chamber was inserted between the stumps, and 7.4 MBq of 131I-NT-4 was administered into the chamber. At the distinct moment of post-injection, the head of one rabbit was imaged at coronary position, the bilateral facial nerve trunk and brain-stem of the others were collected and counted respectively. After 1 mg of brain-derived neurotrophic factor (BDNF) and 7.4 MBq of 131I-NT-4 were administered into the chamber of one rabbit, the rabbit was imaged at coronary position of head at distinct moment. RESULTS: 5.08% of 131I-NT-4 was transported into facial nerve trunk at 4 h of post-injection. 131I-NT-4 presented high peak in facial nerve trunk during the period of 8, 12 h, the peak values were 20.58% and 22.74% respectively. 34.75% and 45.57% of 131I-NT-4 were transported into the brain-stem of experimental side at 8 h and 12 h respectively. The transport of 131I-NT-4 was markedly restrained by BDNF in facial nerve. CONCLUSION: NT-4 displays a receptor mediated retrograde transport in facial nerve. PMID- 12536562 TI - [The effect of ketamine on the expression of HSP70 induced by endotoxin in liver of rat]. AB - OBJECTIVE: To observe the effect of ketamine on the expression of heat shock proteins (HSP70) in liver of rat with septic shock. METHODS: Sixty-four rats were randomly allocated to control group, endotoxin group and ketamine group. The model of septic shock was used to examine the expression of HSP70 in rat liver and the TNF-alpha level of blood at 2 hours after endotoxin treatment and dying time for endotoxin group rats. The survival rates for the three groups were compared. RESULTS: There was a significant increase in the expression of HSP70 and the level of TNF-alpha in the endotoxin group as compared with those of control group (P < 0.05). A significant increase in the expression of HSP70, a decrease in the level of TNF-alpha and a significantly higher survival rate were seen in ketamine group when compared with those of endotoxin group, respectively. CONCLUSION: Ketamine could increase the expression of HSP70 in rat liver, restrain the release of TNF-alpha caused by endotoxin, and raise the survival rates of rats with septic shock. PMID- 12536563 TI - [Pilot study on mutations of p53 gene in laryngeal carcinoma]. AB - OBJECTIVE: To determine the characteristic and frequency of the point mutation of p53 gene. METHODS: 22 cases of laryngeal squamous cell carcinoma (LSCC) with positive expression of mutant p53 protein and 2 cases of normal laryngeal epithelium as control were examined by means of polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP), single strand conformation polymorphism (SSCP) and DNA sequencing analyses. RESULTS: 3 cases of LSCC had the point mutation on codons 248, 250 and 254 of exon 7 in p53 gene. None of the cases had the point mutation on codon 249. CONCLUSION: These findings suggest that the point mutation on codon 249 of exon 7 in p53 gene may be uncommon in LSCC. SSCP and DNA sequencing analyses are sensitive and rapid methods for detecting p53 gene mutation. PMID- 12536564 TI - [p53 mutation in lung cancer in Gejiu and Xuanwei regions of Yunnan Province]. AB - OBJECTIVE: To incidence rates of lung cancer among tin miners in Gejiu and among peasants in Xuanwei are higher than the average incidence rate in Yunnan Province, and the causes of the disease in these two regions have been preliminarily clarified. This study was directed to the molecular biological basis of its pathogenesis. METHODS: Twenty-six specimens of the lung cancers of Gejiu tin miners(GTM) and 15 ones of Xuanwei peasants (XP) were examined by single-strand conformation polymorphism analysis of polymerase chain reaction products. RESULTS: Using PCR primers for the most highly conserved region of the p53 including exons 5-8, we have identified p53 mutations in 13 lung cancer tumor DNA samples of GTM and in 4 ones of XP. Nucleotide sequencing of one of the exon 5 mutations of GTM indicated that there was a C to T transition, which resulted in substitution of Val for Ala at amino acid 135, and there was an AAC to TTC transertion in the exon 7 of XP, which resulted in substitution of Phe for Asn at amino acid 247. CONCLUSION: p53 mutation is a frequent incident in lung cancer of GTM and XP, and two new mutation sites were found in p53 gene, which have not been reported before now. There was no statistically significant correlation between the detection of p53 mutation and the patient's age, sex, mining history and histologic type of the cancer. PMID- 12536565 TI - [Expression of cell cycle associated proteins cyclin D1 and P16 in endometrial carcinoma and the correlation between their expression status]. AB - OBJECTIVE: To study the expression of cell cycle associated proteins Cyclin D1, P16 in endometrial carcinoma and their correlation to clinical parameters, and to assess the correlation between their expression status. METHODS: Immunohistochemical method was used to detect Cyclin D1 and P16 expressions in 64 cases of endometrial carcinoma. RESULT: The positive rate of Cyclin D1 was 54.68%. The Cyclin D1 expression was significantly associated with FIGO stage (P < 0.05). The positive rate of P16 was 53.13% and its expression was related to age, histological grade and FIGO stage (P < 0.05). There was an inverse correlation between Cyclin D1 and P16, r = -0.4007 (P < 0.01). CONCLUSION: Cyclin D1 and P16 had cooperative effect and may play an important role in the development and progression of endometrial carcinoma. PMID- 12536566 TI - [Expression of KDR in medulloblastoma and its clinical implication]. AB - OBJECTIVE: To gain an insight into the possible relationship between the expression of kinase insert domain-containning receptor (KDR) and the prognosis of medulloblastoma. METHODS: Fifty cases of medulloblastoma of the posterior fossa and ten cases of normal cerebellar tissue were studied via a combination of immunnohistochemical staining in formalin fixed paraffin-embedded sections. RESULTS: KDR was expressed weakly in 1 of 10 normal cerebellar tissues, while KDR was either moderately or strongly expressed in most of the medulloblastomas (49 of 50 cases). Moreover, KDR was expressed not only in the endothelial cells of tumor vasculature, but also in tumor cells. Correlation analysis indicated that expression of KDR was correlated significantly with survival time (r = -0.527, P < 0.01). CONCLUSION: KDR was expressed in most of the medulloblastomas, not only in the endothelial cells of tumor vasculature, but also in tumor cells. The expression level of KDR had a negative correlation with the survival time, thus it might be used as one of the prognosticators of medulloblastoma. PMID- 12536567 TI - [Microsatellite instability and loss of heterozygosity in squamous cell carcinoma of the head and neck]. AB - OBJECTIVE: To identify the frequency of loss of heterozygosity (LOH) and microsatellite instability (MI) in Chinese sample of squamous cell carcinoma of the head and neck (SCCHN). METHODS: 18 microsatellite markers were selected and used in the analysis of 30 paired SCCHN for LOH and MI by polymerase chain reaction. RESULTS: In 30 cases of SCCHN, LOH was identified in 12 cases (40%) with at least 1 marker. The prevalence rate of LOH was 0 to 21%, much lower than those reported for most markers. 7 out of the 18 markers (D7S480, D7S522, D9S162, D9S168, D9S304, D9S171 and D17S520) were identified as significant (P < 0.05). However there was no statistically significant correlation between the LOH at these loci and the clinical parameters such as pathological types, tumor size and lymph-node metastasis. MI occurred in only 4 patients, but no MI could be observed using the common criteria for defining MI in two or more markers. CONCLUSION: The most common LOH at D7S480, D7S522, D9S162, D9S168, D9S304, D9S171 and D17S520 might imply the existence of potential tumor suppressor gene of a subset of SCCHN, while MI might not be a crucial event. PMID- 12536568 TI - [Expression and significance of urokinase-type plasminogen activator in medulloblastoma]. AB - OBJECTIVE: To detect the expression and evaluate the clinical significance of urokinase-type plasminogen activator (uPA) in medulloblastoma. METHODS: LSAB (labelled streptavidin biotin method) immunohistochemical technique was applied for detecting the expression of uPA in 50 patients with medulloblastoma, and a follow-up survey with Cox regression analysis was made. RESULTS: The staining for uPA was localized on tumor cells and endothelial cells. Cox regression analysis showed that uPA was an independent prognostic factor affecting survival and it had a negative correlation with the prognosis. CONCLUSION: uPA appears to be an independent marker for predicting the prognosis of medulloblastoma. PMID- 12536569 TI - [Effects of angiotensin II on proliferation and expression of interleukin-6 of human kidney fibroblasts]. AB - OBJECTIVE: To investigate the effects of Angiotensin(Ang) II on the proliferation and expression of Interleukin (IL)-6 in human kidney fibroblasts(KFB). METHODS: The human KFB were cultured and identified, the proliferation of KFB was measured by MTT, and the level of IL-6 was measured by radioimmunoassay(RIA). RESULTS: Angiotensin II could stimulate the proliferation of KFB and enhance the expression of IL-6 in protein level on KFB. CONCLUSION: Angiotensin II can stimulate KFB's proliferation and enhance the expression of IL-6 of KFB. These findings suggest that Ang II might play a part in the mechanisms for modulating tubulointerstitial changes and induceing renal fibrosis. PMID- 12536570 TI - [Relation of plasma glucose levels to serum lipids and apolipoproteins levels in middle and old age males]. AB - OBJECTIVE: To gain an insight into the relation of plasma glucose levels to lipids and apolipoproteins levels. METHODS: Fasting 12-14 h blood glucose (BG), high density lipoprotein cholestrol(LDL-C), apolipoproteins(apo) A I, A II, B100, C II, C III and E levels were measured in 735 men aged 40-70 in Chengdu area. The subjects studied were divided into three groups by the plasma glucose levels: 1. normal group (S1), 2. decreased glucose tolerance group (S2), 3. diabetes mellitus (DM) group (S3). RESULTS: Among 735 cases studied, groups S2 and S3 had more hyperlipidemia than group S1 (64%, 45% and 32.2% respectively). The fasting serum mean TG levels in groups S2 (3.62 +/- 2.52 mmol/L) and S3 (4.45 +/- 3.82 mmol/L) group were significantly higher than that in group S1 (1.92 +/- 1.39 mmol/L) (P < 0.01 and P < 0.01), and the mean TG levels in groups S2 (5.68 +/- 1.15 mmol/L) and S3 (5.68 +/- 1.12 mmol/L) were higher than that in group S1 (5.13 +/- 1.06 mmol/L) (P < 0.05, P < 0.05). The serum apo B100, C II, C III and E levels in groups S2 and S3 were significantly higher as compared with that in the normal group (P < 0.01, P < 0.05). The correlation analysis indicated that there was a positive correlation of BG with serum TG in groups S1, S2 and S3 whose correlation coefficients (r) were 0.1800, 0.1436 and 0.3212 respectively (P < 0.001); and a negative correlation with HDL-C levels in groups S2 and S3 (r = 0.2534 and -0.1477, P < 0.001). The plasma glucose levels were positively correlated with apo B100, C III and E levels in groups S1, S2 and S3, P < 0.001), with apo C II in groups S1 and S2 (P < 0.001, P < 0.01); and negatively correlate to apoA I only in group S2 (r = -0.1182, P < 0.01). CONCLUSION: The results suggest that serum TG, TC, HDL-C, apoB100, C II, C III and E levels be the main factors that related to plasma glucose levels. PMID- 12536571 TI - [The paraoxonase Gln-Arg192 polymorphism in patients with coronary heart disease in Chinese population]. AB - OBJECTIVE: To investigate the 192 Gln-Arg polymorphism of paraoxonase (PON) gene and its relationship with serum lipids levels in patients with coronary heart disease in Chinese population. METHODS: The genotype and allele frequency of paraoxonase gene 192 Gln-Arg polymorphism was assayed by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP). Serum lipids were measured by enzymatic kits in 118 CHD patients and 128 healthy subjects whose fasting serum TG levels were < 1.82 mmol/L and TC levels < 6.2 mmol/L from a population of Chinese Han nationality in Chengdu area. RESULTS: Both in CHD group and control group, the QR genotype of PON gene was the major one, and its frequencies were 0.578 and 0.525 respectively. No differences were found in PON gene Gln-Arg polymophism in the CHD group when compared with the control group. The frequency of R allele of PON gene in Chinese was significant higher than that in European and American Caucasians (0.51 vs. 0.26-0.31, P < 0.01) and Indian (0.51 vs. 0.17, P < 0.01), and significantly lower than that in Japanese (0.51 vs. 0.65, P < 0.01). CONCLUSION: These results suggested that the Gln-Arg192 polymorphism of the PON1 gene was not associated with coronary heart disease in Chinese population. PMID- 12536572 TI - [Serum apolipoprotein A I, B100 and E levels and apolipoprotein E polymorphism in patients with Alzheimer's disease and multiple infarction dementia in Chinese population]. AB - OBJECTIVE: To investigate the relationship of serum apolipoprotein (apo) A I, B100 and E levels and apo E polymorphism to Alzheimer's disease (AD) and multiple infarction dementia (MID) in Chinese population. METHODS: The apoE phenotypes were assayed by isoelectric focusing and immunoblotting, and apolipoproteins were determined by radial immunodiffusion assay in 75 patients with AD, 36 patients with MID and 60 control subjects. RESULTS: The frequency of apo E4 allele (epsilon 4) was significantly higher and the frequency of epsilon 2 allele was lower in AD group, compared with those in the control group (0.2333 vs 0.0666, 0.0467 vs 0.0833, P < 0.05). The frequency of epsilon 4 allele in MID group was also higher than that in the control group (0.2083 vs 0.0666, P < 0.05). The fasting serum apoA 1 and E levels in AD and MID groups were remarkably lower than those in the control group (P < 0.001 and P < 0.05). CONCLUSION: Apo E4 allele epsilon 4 was associated with AD and MID. ApoE4 might be a risk factor for AD and MID, and apo E2 might be a protective factor for AD. The serum apo A I and apoE levels were significantly decreased in patients with Alzheimer's disease and multiple infarction dementia. PMID- 12536573 TI - [Appraisal of occupational stress and strain in primary and secondary school teachers]. AB - OBJECTIVE: This study was conducted to assess occupational stress and strain in primary and secondary school teachers. METHODS: A test of occupational stress and strain was carried out by using Occupational Stress Inventory Revised Edition (OSI-R) in 1460 primary and secondary school teachers (teacher group) and 319 mental workers in non-educational area (non-teacher group as control). RESULTS: The results showed the level of occupational stress in role overload and physical environment in the teacher group was significantly higher than that in the non teacher group (P < 0.05). In teacher group the level of occupational stress and strain increased with age; the occupational stress and strain in male teachers were significantly higher than those in female teachers (P < 0.01); the occupational stress and strain in secondary school teachers were significantly higher than those in primary school teachers. CONCLUSION: These results indicate: to protect and promote primary and secondary school teacher's health, particularly male teachers' health, to mitigate their work pressure and to raise the quality of education are important tasks in the area of occupational health. PMID- 12536574 TI - [Assessing health state utility using the short form 36]. AB - OBJECTIVE: To explore the methods of utility assessment using the short form 36. METHODS: The authors selected 70 people stratified by age, sex, and occupation in a city in Sichuan province, and investigated the preference of the population on each domain or item by paired comparison with other domains or items. Based on the preference, the weights of the domains and items were calculated. RESULTS: The weights of all the 8 domains and the items within 7 domains except the first domain "physical functions" were identified. The total score and the scores of the domains calculated by weighted adding method were significantly different from those calculated by simple adding method. However, only little variation of the scores of the domains between the two methods appeared. CONCLUSION: Utility calculated by weighted adding method may reflect people's preference to the health states more accurately. However, the value is limited in using weighted adding method for calculating the scores of domains. PMID- 12536575 TI - [The influence of somatostatin 14 on the proliferation of human retroocular fibroblasts]. AB - OBJECTIVE: To elucidate the possible mechanism of Somatostatin 14 in the treatment of Graves' ophthalmopathy, thus providing laboratory data with the intent to underpin therapeutic developments. METHODS: The human retro-ocular fibroblasts (HROFb) was cultured and MTT colorimetry assay was used to determine the proliferation of HROFb. RESULTS: MTT colorimetry assay could precisely reflect the proliferation of HROFb. The absorbance was directly proportional to the numbers of the cells (r = 0.996, P < 0.001). Somatostatin 14 could enhance the cell proliferation percentage at lower levels (< 25 nmol/L) and inhibit that at higher levels (> 25 nmol/L). The cell proliferation correlated positively to Somatostatin 14 levels significantly (r = -0.9054, P < 0.001). CONCLUSION: Somatostatin 14 has a dual effect on HROFb. PMID- 12536576 TI - [Studies on insulin-like growth factor I receptor in fetal rats with intrauterine growth retardation]. AB - OBJECTIVE: This study was conducted to detect the localization of the insulin like growth factor I receptor (IGF-I R) in the cells and tissues in the rats with intrauterine growth retardation (IUGR) and normal growth rats. METHODS: 21 pregnant rats were randomly divided into the experimental group (IUGR) (n = 12) and the control group (n = 9). Animal models for IUGR were established by clamping the uterine vasculature of pregnant rats for 20 minutes on day 14 of gestation. The control group rats were subjected to Sham-operation. On day 22 of gestation, fetal rats were delivered by cesarean section. Immunohistochemical assay was performed to detect the IGF-I R expression on liver and lung using specific polyclonal antibodies to rat IGF-I R. RESULTS: In the experimental group, the IUGR rats' body weight, height, and the weight of their liver, lung and the placenta were significantly lower than those in the control group. In the IUGR rats' liver, the area ratio of IGF-I R was increased and the average gray level was decreased, compared with those in the control group. In the IUGR rats' lung, the area ratio of IGF-I R was increased, compared with that in the control group, but no difference of average gray level was observed between the two groups. CONCLUSION: The increased expression of IGF-I R in rats with IUGR, may be due to the decrease of IGF-I and the compensative mechanism of the body. PMID- 12536577 TI - [The murine submandibular gland conditioned medium induce the development of CFU GM and CFU-meg]. AB - OBJECTIVE: Our purpose is to determine whether murine submandibular gland produce hematopoietic growth factors or not and which hematopoietic growth factor are produced by murine submandibular gland. METHODS: By using the techniques of culture of hematopoietic progenitor cells(CFU-GM, CFU-Meg) ex vivo, flow cytometry, the effects of murine submandibular gland conditioned medium(SGCM) on the development of CFU-GM and CFU-Meg in mice was studied. RESULTS: SGCM can stimulate the development of CFU-GM and CFU-Meg (without any exogeneous HGF), almost no colony in control (without any exogeneous HGF and SGCM); the promoting activity of male SGCM is higher than female SGCM's in augmenting colony formation of CFU-Meg (P < 0.05), there is no difference of CFU-GM(P > 0.05). The stimulating activity of anemic mice SGCM is higher normal mice SGCM's (P < 0.05). IL-3 can collaborate with female SGCM to stimulate proliferation of CFU-Meg. SGCM can promote bone morrow cell entiring the active proliferative phase(S/G2). CONCLUSION: Submandibular gland can stimulate the development of CFU-GM and CFU Meg by secreting hematopoietic growth factor-like activities. Our study provide the experimental evidences for clarifying the relationship between submandibular gland and hematopoiesis. PMID- 12536578 TI - [The effect of rIFN alpha-2b on mice NK cell activity]. AB - OBJECTIVE: This animal experiment was made to study the effect of the recombined interferon alpha-2b(rIFN alpha-2b) on natural killer cells activity of mice spleen in vivo, and by way of this, to test the activity of rIFN alpha-2b. METHODS: BALB/c mice were given rIFN alpha-2b subcutaneously. The inhibiting 3H TdR intaking method was used to test the NK activity of spleen cells of BALB/c mice. RESULTS: The NK cell activity levels of the high-dose group, the mid-dose group and the low-dose group were 65.19 +/- 17.56, 44.19 +/- 14.30 and 35.46 +/- 17.49, respectively. CONCLUSION: rIFN a-2b can enhance NK cells activity observably and its effect is dose dependent. PMID- 12536579 TI - [The effect of ETA receptor antagonist WS009A on the pathogenesis of hypoxic pulmonary hypertension in rats]. AB - OBJECTIVE: To investigate the effect of ETA receptor antagonist, WS009A, on the pathogenesis of hypoxic pulmonary hypertension in rats. METHODS: In the course of chronic hypoxia, rats were treated with WS009A (10 mg/kg) by a siliastic catheter inserted in the left external jugular vein. RESULTS: The measured values of mean pulmonary arterial pressure (mPAP) were 2.7 +/- 0.33, 3.97 +/- 0.47 [symbol: see text] 2.39 +/- 0.27 kPa in the control, the hypoxic and the WS009A treated groups, respectively. The percentages of vascular wall area/total vascular area (MA%) were 0.26 +/- 0.03, 0.52 +/- 0.04, 0.32 +/- 0.07, respectively. The percentages of vascular wall thickness/vascular external diameter(MT%) were 0.14 +/- 0.02, 0.31 +/- 0.03, 0.18 +/- 0.05, respectively. The ratios of right ventricular wall to left ventricle plus septum[RV/(LV + S)] were 0.24 +/- 0.02, 0.35 +/- 0.03, 0.26 +/- 0.03, respectively. The variance analysis revealed there were decreases in mPAP and the ratio of RV/(LV + S) in WS009A group (P < 0.01, compared with those in hypoxic group). There were no haemodynamic and right ventricular hypertrophy differences between WS009A group and control group. The MT% and MA% in WS009A groups were higher than those in controls, but there were significantly decreased MT% and MA% in WS009A group as compared with those in hypoxic group (P < 0.01). CONCLUSION: The ETA receptor antagonist, WS009A, attenuates the pulmonary vascular wall remodelling and the pulmonary hypertension induced by chronic hypoxia. PMID- 12536580 TI - [A study of the acting mechanism of aspirin for resistance to oxidative damage]. AB - OBJECTIVE: To explore the mechanism of aspirin for resistance to oxidative damage in endothelial cells. METHODS: Using cultured endothelial cells, we measured the levels of aspirin induced ferritin expression on resistance to hydrogen peroxide toxicity toward cells in the presence of the iron chelator desferrioxamine added to FeCl3. RESULTS: Aspirin at low concentration (0.1 mmol/L) induced significant increase of ferritin expression in a time- and concentration-dependent fashion up to 25% over basal levels(P < 0.05). Preincubating the cells for 8 h with aspirin (0.1 mmol/L) reduced lactate dehydrogenase(LDH) release rate by 50%, toxicity reduction by 40%, and significant decrease of malondialdehyde (MDA) production. Aspirin induced cytoprotection from H2O2 damage was also in a concentration- and time-dependent fashion. However, in the presence of the iron chelator desferrioxamine, aspirin enhanced ferritin synthesis was abrogated, in contrast, FeCl3 increased aspirin induced ferritin synthesis in cells. CONCLUSION: The study suggested that the antioxidation of aspirin was brought into action by affecting the cellular iron metabolism pathway to induce ferritin synthesis. PMID- 12536581 TI - [Protective effect of berberine on isolated perfused heart in heart failure]. AB - OBJECTIVE: To examine the protective action of berberine against the development of heart failure. METHODS: Wister rats were divided into two groups. The Langendorff perfusion of isolated heart was performed and Verapamil was used to bring about acute heart failure. The experiment group was given berberine (10(-6) mol/L) before the use of Verapamil, but the control group was not given the berberine. A comparison was made on the degree of heart failure between the two group. RESULTS: The degree of heart failure in the experiment group was significantly less severe than that in the control group (P < 0.001). CONCLUSION: Berberine has the protective action against to the development of heart failure. PMID- 12536582 TI - [The preparation of collagen burn pellicle of compound sulfadiazine silver and assessment of its efficacy in an animal experiment on deep partial thickness burn wound]. AB - OBJECTIVE: To prepare the collagen burn pellicle of compound sulfadiazine sliver and observe its therapeutic effect on deep partial thickness burn wound. METHODS: The initator method was adopted for preparing the collagen burn pellicle of compound sulfadiazine sliver. A model of deep partial thickness burn wound was established in 84 SD rats for the observation of the effect of collagen burn pellicle of compound sulfadiazine sliver on burn wound healing. RESULTS: The collagen burn pellicle of compound sulfadiazine sliver enhanced the coming off of necrotic tissues and the healing of burn wound. The hydroxyproline content of burn wound was higher in the experiment group than that in the comparison groups, (P < 0.05). The percentage of G0/G1-phase in full skin cells of burn wound at 5, 7, 10 and 14 days after burn was lower than that in the comparison groups (P < 0.05). The percentage of S-phase at 5, 7 and 14 days was higher in the experiment group than that in the comparison groups (P < 0.05). The water content in full skin cells of burn wound at 24, 36 and 48 hours after burn was significantly lower in the experiment group than that the comparison groups (P < 0.05). CONCLUSION: The collagen burn pellicle of compound sulfadiazine sliver can enhance wound healing in the management of deep partial thickness burn wound. PMID- 12536583 TI - [The preparation and study on hepatic targeting tendency of galactosyl-anti-CD3 McAb in mice]. AB - OBJECTIVE: This study sought to reduce the recurrence rate of primary liver cacer (PLC) after hepatectomy by increasing the concentration of infiltrating lymphocytes(TILs) at the target organ. METHODS: It has been reported that galactosyl-anti-CD3-McAb has an obvious hepatic targeting tendency in vitro. On the basis of that work, the present authors conducted a study in mice. Galactosyl anti-CD3-McAb was prepared and its carbohydrate density was measured with the phenol-sulphyric acid method, and then anti-CD3-McAb (labeled with 125I) and galactosyl-anti-CD3-McAb (labeled with 131I) were infused respectively via the peripheral vein and the radioactivity in each organ was measured. RESULTS: The carbohydrate density of galactosyl-anti-CD3-McAb in this experiment was 58.12, which effectively guaranteed the specific binding between galactosyl-anti-CD3 McAb and hepatic binding protein(HBP). It was also noticed that anti-CD3-McAb tended to aggregate in the lungs while being infused via the peripheral vein, and that galactosyl-anti-CD3-McAb had an obvious hepatic targeting tendency in vivo and it stayed in liver for quite a long period. CONCLUSIONS: Galactosyl-anti-CD3 McAb has an obvious hepatic targeting tendency in vivo while being infused through peripheral vein; this may be due to the specific binding between galactose and HBP. PMID- 12536584 TI - [Study of peak expiratory flow in 728 normal adolescents in Chengdu area]. AB - OBJECTIVE: This study was carried out to build the normal values and regression equations of peak expiratory flow (PEF) in normal adolescents. METHODS: PEF and routine ventilatory functions, FVC, V75, V50, V25 were measured in 728 normal adolescents aged 8-19. RESULTS: The PEF and FVC, V75, V50, V25 rose with the increase of age. The regression equations showed that PEF was highly positively correlated with age and FVC both in male and female adolescents, and also positively correlated with body weight in males; the coefficients of multiple correlation were 0.85 and 0.78 in male and female ones respectively. Correlations of PEF with V75, V50, V25 were also high, especially with V75 of which the correlation coefficients came up to 0.98 both in male and female adolescents. The coefficients of variation were 15%-20% both in male and female adolescents, and were in normal ranges. CONCLUSION: The normal ranges and regression equations of PEF provided for normal adolescents can be used to evaluate the pulmonary functions of patients who are in the same age-group suffering from bronchial asthma and other respiratory diseases. PMID- 12536585 TI - [Measurement of the parameters of dose-response curve obtained after the methacholine challenge test]. AB - OBJECTIVE: To investigate the clinical value in the measurement of parameters of the dose-response curve (DRC) obtained after the methacholine challenge test. METHODS: Twenty-seven cases of cough variant asthma (CVA), 29 of mild asthma, 19 of moderate asthma, and 15 healthy volunteers underwent the methacholine challenge test. The dose-response curves were constructed. The following parameters were calculated: position (PC35sGaw), dose-response curve (DRS), ratio of the area under the curve to the logarithm of the maximal concentration (AUC/lg[Cmax]) and maximal response. RESULTS: A plateau appeared in 11 cases among the healthy subjects, while a plateau appeared only in 2 cases of CVA and 2 of asthma. There were no significant differences among the asthmatic groups in the measurements of PC35sGaw and AUC/lg[Cmax] (geometric mean PC35sGaw: mild asthmatics = 0.369, moderate asthmatics = 0.251, cough variant asthmatics = 0.547), AUC/lg[Cmax] was significantly greater in the asthmatic groups than in the normal group [AUC/lg [Cmax](mean +/- s): mild asthmatics = 24.7 +/- 4.7, moderate asthmatics = 26.6 +/- 4.3, cough variant asthmatics = 25.6 +/- 3.7, normal subjects = 15.5 +/- 4.3, P < 0.01]. DRS of moderate asthmatics was significantly greater as compared with cough variant asthmatics (geometric mean DRS: mild asthmatics = 30.761, moderate asthma-tics = 59.020, cough variant asthmatics = 19.231, P < 0.05), but there was no difference between the other groups. PC35sGaw was negatively correlated with DRS (r = 0.866, P < 0.001) and with AUC/lg[Cmax] (r = 0.502, P < 0.001). CONCLUSION: Four parameters of DRC obtained after the methacholine challenge represent airway sensitivity and reactivity respectively. The parameters of DRC of asthmatic patients are different from those of normal subjects. The sensitivity is related to the reactivity, but they are not completely parallel with each other. PMID- 12536586 TI - [The levels of angiotensin and aldosterone in gases of hyperthyroid Graves' disease with and without hypokalemic periodic paralysis]. AB - OBJECTIVE: To investigate the relationships between the changes of plasma angiotensin I (AT I). Angiotensin II (AT II). serum aldosterone (Ald) levels and hypokalemic periodic paralysis (THPP) complicating hyperthyroid Graves' disease. METHODS: The levels of serum potassium, TT3, TT4, FT3, FT4, Ald and the plasma AT I, AT II and 24 hours urine potassium output were determined in 73 cases of hyperthroid Graves' disease, of which 43 were complicated by THPP and the other 30 were grouped as NTHPP cases. RESULTS: The mean level of serum potassium in THPP group was significantly lower than that in NTHPP group (P = 0.000), and the mean level of 24 hours urine potassium output in THPP group was significantly higher than that in NTHPP group(P = 0.000); The mean levels of plasma AT II and serum Ald in THPP group in upright position were significantly higher than those in NTHPP group (P = 0.008 and 0.013, respectively). The mean levels of 24 hours urine potassium output, plasma AT II and serum Ald in upright position were negatively correlated to serum potassium in both groups. The level of serum Ald in upright position was positively correlated to the level of 24 hours urine potassium out-put. The mean levels of plasma AT I and AT II in supine position were positively correlated to serum T3, and the level of plasma AT II in upright position was positively correlated to serum FT3. CONCLUSION: The level of 24 hours urine potassium output rises in THPP patients; the high levels of AT and Ald may contribute to the pathogenesis of the disease. PMID- 12536587 TI - [Research on the high strength all ceramic crown by the combination of CAD/CAM and glass infiltrated alumina blocks]. AB - OBJECTIVE: To search for a method that fabricates the high strength all ceramic crown by the combination of CAD/CAM and glass-infiltrated alumina. METHODS: Alumina coping was milled by the Cerec II CAD/CAM and glass was infiltrated into alumina by the capillary effect. Vita alpha ceramic was used to veneer the surface of substructure for all ceramic crown. RESULTS: This technology need not duplicate die and sinter alumina substructure for 10 h, compared with the conventional In-ceram technology. The time of glass infiltration has been decreased from 4 h to 40 minutes by the high capillary effects of prefabricated alumina blocks. CONCLUSION: This is a new method for all ceramic crown by using Cerec II CAD/CAM to mill a simple ceramic block so as to fabricate prosthetics in clinic. PMID- 12536588 TI - [A study on the bond interface between low-fusing dental porcelain and pure titanium]. AB - OBJECTIVE: To evaluate the bond interface between low fusing dental porcelain and pure titanium by observing the topography and detecting the ionic diffusion in the interface area. METHODS: The low fusing-porcelain La-porcelain produced by the authors or Vita Titankeramik porcelain was fused to the surfaces of pure titanium. The topography of the interface between pure titanium and porcelain, and the structure of experimental materials were observed with SEM. The state of ionic diffusion in the interface area was investigated with EPMA. RESULTS: Excellent permeation and diffusion of La-porcelain were observed on the surfaces of pure titanium. The diffusion of ions of stannum and silicon was discovered in the interface area. The microstructure of La-porcelain to pure titanium bond interface was finer than that of Vita Titankeramik porcelain. CONCLUSION: Excellent bond can be produced in the interface between La-porcelain and pure titanium. The bonding mechanism may involve mechanical bond and chemical bond. The ionic diffusion of stannum plays an important role in the bonding of porcelain to pure titanium. PMID- 12536589 TI - [Detection of interleukin-6 in gingival crevicular fluid from active and non active adult periodontitis sites]. AB - OBJECTIVE: To reveal the relationship between IL-6 and the activity of periodontitis. METHODS: The concentration of IL-6 in gingival crevicular fluid (GCF) was measure by ELISA. Sixty teeth from 39 patients with adult periodontitis were sampled, among which there were 29 teeth in active phase, while the rest were in non-active phase. RESULTS: Significant difference was found in the level of IL-6 in GCF between the active group (10.65 +/- 22.05 ng/ml) and the non active group (1.73 +/- 0.53 ng/ml, P < 0.05). CONCLUSION: The level of IL-6 in GCF may be a useful indicator of the activity of periodontitis. PMID- 12536590 TI - [The measurement of 153Sm-EDTMP bone uptake rate and its relationship with therapeutic effect]. AB - OBJECTIVE: To evaluate the measurement of 153Sm-EDTMP Bone uptake rate with whole body scintigraphy and analyze the relationship between bone uptake rate and therapeutic effect. METHODS: 66 patients with bony metastases were examined with whole-body scintigraphy 10 minutes and 6 hours after the administration of 153Sm EDTMP, and then bone uptake rate was calculated. RESULTS: The bone uptake rate in 66 patients ranged from 31.9% to 86.6%, (mean 56.0%). The patients with higher bone uptake rate had better overall therapeutic effect. According to the bone uptake rate calculated by whole-body scintigraphy, the therapy activity administered within the limit of bone marrow absorbed dose of 1400 mGy in these patients was 1.40-2.27GBq (mean 1.90 GBq). CONCLUSION: The present authors have developed a whole-body scintigraphic technique which is simple and reliable in calculating prospectively a safe and effective dose of 153Sm-EDTMP for palliating the patient's painful bone metastases without myelotoxicity. PMID- 12536591 TI - [A study comparing 131I versus 131I plus antithyroid drug in the management of Graves' disease]. AB - OBJECTIVE: To ascertain whether the therapoutic effect of 131I is affected by the additional use of thyrostatic medication. METHODS: One hundred and eighty-seven patients with Graves' disease (GD) were randomly assigned to treatment with 131I alone or plus thyrostatic medication(PTU) 3 days after the beginning of 131I therapy. All patients were examined every month for six months after treatment. RESULTS: The cure of hyperthyroidism occurred in 71 of 93 patients(76.3%) treated with 131I alone and in 76 of 94 patients(80.5%) treated with 131I plus thyrostatic medication(PTU). There were no significant differences in cure rate between the two groups. CONCLUSION: The additional use of thyrostatic medication 3 days after the beginning of 131I therapy will not affect the efficacy of 131I in the treatment of GD. PMID- 12536592 TI - [Clindamycin soluble suppository-production and its efficacy in treating bacterial vaginosis]. AB - OBJECTIVE: To make the soluble suppository of clindamycin which is an effective drug for treatment of Bacterial vaginosis and to treat patients in clinical trials. METHODS: Outpatients who had vaginal infections from Sept. 1998 to Jan. 1999 were investigated. The diagnoses of bacterial vaginosis were made according to Amsel's criteria and 80 patients were included in this study. They were divided into two groups to be treated respectively by metronidanole suppository and clindamycin suppository which were made by our hospital pharmacy. RESULTS: The cure rate of metronidanole was 87.5% and that of clindamycin 90.3%; No significant difference was noted between the two groups(P > 0.05). The new clindamycin suppository was not irritant and it was easy to dissolve in vagina. There was no discontinued treatment because of side-effect. CONCLUSION: The clindamycin suppository does not irritate the mucosa of vagina, it is an efficacious remedy for bacterial vaginosis. PMID- 12536593 TI - [Quantitative detection of flow indices in tubal flow via pulsed Doppler ultrasound, in vitro]. AB - OBJECTIVE: This paper deals with the method and feasibility of quantitative detection of flow indices via commercially available Pulsed Doppler Ultrasound. METHODS: The indices, such as Reynolds number, relative spectral broadening (RSB), laminar shear stress and Reynolds normal stress (RNS) were selected and evaluated. Detection was done temporally and spatially, upstream and downstream of the stenosis. The data were evaluated qualitatively and quantitatively. RESULTS: The roughest turbulence was met at 1/2R sample volume, 1.0 cm downstream of the stenosis. CONCLUSION: The methodology in this study is proved to be feasible and may have good prospects in clinical use. PMID- 12536594 TI - [Determination of paraoxonase activities in serum and HDL]. AB - OBJECTIVE: [corrected] To establish a reliable method of determining the activity of paraoxonase(PON) in serum and high density lipoprotein(HDL). METHODS: We used phenylacetate(PA) as substrate and investigated the hydrolysis of PA catalyzed by serum PON or HDL-PON. The PON activities were calculated from the velocities of reaction that had been determined by the increasing absorbance of product p phenol within the first 5 minutes. RESULTS: Through the studies on the effects of substrate concentration, pH, activator Ca2+, and inhibitor EDTA, we found the best conditions of reaction to be: substrate concentration 5 mmol/L, pH8.0, and Ca2+ concentration 2 mmol/L. And we established the method with good reproducibility and stability. CONCLUSION: Under the above-stated conditions, it is reliable to determine the activity of paraoxonase in serum and HDL. PMID- 12536595 TI - [The millipore rapid screening of hydantoinase production strains]. AB - OBJECTIVE: To find a strain with a higher hydantoinase activity. METHODS: The millipore rapid screening method was established to select the strains. The effects of several solvents on the rate of biotransformation were compared. RESULTS: A hydantoinase production strain has been selected by the millipore rapid screening method. Tween-80 (1%) is found to be a suitable solvent to dissolve the substrate. CONCLUSION: A new method is provided for rapid selection of hydantoinase production strains from large-scale samples. The difficulty in dissolving the substrate has been resolved to some extent. PMID- 12536596 TI - [Determination of trace amount of cadmium with vapor generation and atomic absorption spectrometry]. AB - OBJECTIVE: This study aimed to establish a sensitive method for the determination of trace amount of cadmium. METHODS: The reaction systems for the generation of volatile cadmium species were investigated with vapor generation and atomic absorption spectrometry. RESULTS: It was found that potassium ferricyanide with hydroxylamine hydrochloride greatly enhanced the generation efficiency of the volatile species. The reaction parameters using ferricyanide-hydroxyamine tetrahydroborate in acidic acqueous solution were optimized for efficient generation of volatile cadmium species. A detection limit (3 sigma blank, peak height mode) of 11 pg/ml was obtained under optimized conditions. The RSD of the determinations for 0.2 ngCd/ml was 3.4%. The effects of 33 diverse ions on the generation of the cadmium volatile species were investigated. The method was applied to the determination of cadmium in standard reference materials, the results obtained were in good agreement with the certified values. CONCLUSION: The method can be applied to the determination of trace cadmium in environment and food samples. PMID- 12536597 TI - [Determination of 35 polycyclic aromatic hydrocarbons in airborne suspended particulate by GC/MS]. AB - OBJECTIVE: To establish capillary column GC/MS method for the determination of 35 polycyclic aromatic hydrocarbons (PAHs) in airborne suspended particulate. METHODS: The air samples collected by an electrostatic particle collector were Soxhlet-extracted using 3:1 benzene/ethanol, and then subjected to the procedures of concentration and clean-up. Finally, the target PAHs were determined by the proposed method. RESULTS: All the target PAHs could be well separated except for benzo[b] fluoranthene, benzo[j] fluoranthene and benzo [k] fluoranthene through temperature programming. The relative standard deviations of the method were 1.8% 17.7% and the recoveries ranged from 93.4% to 101.9%. The absolute detection limit ranged from 0.0001 ng to 0.1 ng. CONCLUSION: The method is applicable to the determination of more than 30 PAHs in the airborne suspended particulate samples. PMID- 12536598 TI - [Establishment and implication of an assay for high density lipoprotein phospholipids in human serum]. AB - OBJECTIVE: To develop an assay for high density lipoprotein phospholipids in human serum based on ascorbutate reduction method. METHODS: HDLs were separated from apolipoprotein B-containing lipoproteins by precipitation of phosphotungstic acid and magnesium chloride. Phospholipids of HDL were extracted by ethanol/ether, and dried. After the dried phospholipids were digested by sulphuric acid and perchloric acid, the color was developed by adding ammonium molybdate in ascorbutate. The levels of high density lipoprotein phospholipids (HDL-PL) were measured by spectrophotometry at 700 nm. RESULTS: The coefficients of variation (CV) were 3.6% and 3.7% within two batches of assays. Recovery of isolated HDL-PL added to serum ranged from 98% to 107%, averagely 103%. The established assay for human serum HDL-PL was used to measure the serum levels of 30 hypercholesterolemic subjects, 30 hypertriglyceridemic subjects, 30 combined hyperlipidemic subjects, and 30 normolipidemic subjects. The hypertriglyceridemic subjects had lower HDL-PL level than normolipidemic subjects and hypercholesterolemic subjects (The P values are 0.005 and 0.007 respectively). CONCLUSION: A simple and specific method for assay of HDL-phos-pholipids in human serum has been developed. The above data collected by the use of this method demonstrate the closer relationship between human HDL-PL metabolism and triglyceride metabolism, suggesting that lower HDL-PL level might serve as an index in the assay for type IV hyperlipidemia. PMID- 12536599 TI - [Counting platelets by flow cytometry with platelet-specific antibody CD42a]. AB - OBJECTIVE: To investigate a reference method for platelet (PLT) count. METHODS: 76 samples of daily whole blood of high, normal and low values in patients were collected and analyzed by flow cytometry (FCM) with CD42a monoclonal antibody that could combine specifically with the platelet membrane glycoprotein Ib-Ix (GPIb-Ix). PLT counts were analyzed by multiplying PLT/RBC ratio from FCM with the RBC that was counted by a calibrated blood analyzer. Meanwhile, the results of PLT from FCM were compared with that from blood analyzers SE-9500, NE-1500, CD 1600 or manual count. RESULTS: The within-replicate coefficient of variance(CV) of FCM was less than 4% for a high count(501 x 10(9)/L) and middle count(184 x 10(9)/L), the CV of low count(23 x 10(9)/L) was 11.432%, the between-replicate CV was 10.792%, and the overall reproducibility was less than 5%. Linearity was demonstrated well(r = 0.9975 P < 0.001) when the PLT counts were 20 x 10(9)/L-700 x 10(9)/L. PLT count was stable at room temperature within 6 hr. The carry-over was 0.87%. The results between FCM and SE-9500, NE-1500, CD-1600 or manual count was comparable (r > 0.975, P < 0.001). CONCLUSION: These findings confirm that FCM for PLT count is specific, accurate and precise and could be considered as a candidate of reference method for PLT count. PMID- 12536601 TI - [Construction of three-dimensional finite element model of rat molar]. AB - OBJECTIVE: This paper was to construct three-dimensional finite element model of rat molar, and to access mechanical stress distribution in the tooth and periodontia. METHODS: The first maxillary molar of adult rat was moved for 2 weeks with initial force of 40 g and 100 g. The histological specimen of tooth and surrounding structures were sectioned serially at 4 microns-thick slices in the coronary direction. The section's microscopic photographic images were made every 140 microns. The structural information of images was traced and fed into computer for three-dimensional reconstruction of the structure and finite element model. RESULTS: The stress distribution in periodontia which lies between tooth and alveolar bone was very complicated. CONCLUSION: The three-dimensional finite element model of rat molar is simple, precise, and effective for analyzing stress in tooth and periodontia. PMID- 12536600 TI - [Effects of heat shock pretreatment on the translocation and expression of alpha B-crystallin in cultured neonatal rat cardiomyocytes]. AB - The effects of heat shock pretreatment on translocation and expression of alpha B crystallin were evaluated in neonatal rat cardiomyocytes. RESULTS: 1. Western blot analysis demonstrated that cytosolic soluble alpha B-crystallin rapidly translocated to insoluble intracellular structure at 10 or 30 minutes and returned to cytosolic soluble compartment at about 1-2 h after heat shock. 2. Double staining of immunofluorescence showed that alpha B-crystallin redistributed in cytosol after heat shock, while HSC70 translocated from cytosol to nucleus. 3. Quantitative analysis with immunoelectronic microscopy demonstrated alpha B-crystallin translocated to cytoskeletal structure such as Z line and nuclear envelope after heat shock. 4. Northern-blot showed that heat shock induced alpha B-crystallin mRNA expression. The rapid translocation and increasing expression of alpha B-crystallin after heat shock suggests that this small heat shock protein may play an important protective roles during the early and late phases of myocardial ischemic preconditioning. PMID- 12536602 TI - [Experimental study of low-molecular weight heparin on ameliorating progressive glomerulosclerosis]. AB - OBJECTIVE: This article was to study the effect and mechanism of low-molecular weight heparin (LMWH) on ameliorating progressive glomerulosclerosis. METHODS: The progressive glomerulosclerosis models were established by repeated injection of adriamycin to the rats after the removal of their right kidneys. The therapeutic group was received the treatment of LMWH for six weeks. The proteinuria, blood-biochemistric, and renal pathologic lesion were investigated respectively; the sclerotic index of glomerulus were calculated and the expression of collagen IV (Col-IV), laminin, platelet-derived growth factor B(PDGF-B) and basic fibroblast growth factor (bFGF) in glomerulus were determined by immunohistochemistry simultaneously. RESULTS: LMWH not only reduced the proteinuria, elevated the serum albumin, improved the renal function, alleviated the renal pathologic lesion, and reduced the sclerotic index of glomerulus, but also significantly decreased the accumulation of Col-IV and laminin and the expression of PDGF-B and bFGF in glomerulus. CONCLUSION: LMWH may obviously ameliorate the progressive glomerulosclerosis; the decreased accumulation of mesangial matrix by inhibiting the expression of PDGF-B and bFGF in glomerulus may be one of the important mechanisms. PMID- 12536603 TI - [Effect of activation of p-MAPK on activating c-fos and c-jun proteins in breast cancer]. AB - OBJECTIVE: This paper was to investigate the relationship between expression of p MAPK and oncogenesis of breast cancer. METHODS: Immunohistochemical technique was used to detect the expression of p-MAPK and c-fos and c-jun proteins in 68 cases of breast cancers, 42 cases of pericarcinomatous tissues and 7 cases of normal breast tissues. RESULTS: Positive stainings of p-MAPK, c-fos, and c-jun were localized in cancer cell nuclei. The positive rates of p-MAPK, c-fos, and c-jun were 86.8% (59/68), 82.4% (56/68), and 77.9% (53/68), respectively, which were much higher than that in pericarcinomatous tissues (P < 0.01). Of some cases, p MAPK positive staining was also found in the nuclei of the fibroblastic and angioendothelial cells of cancer stroma. CONCLUSION: Activated or overexpressive MAPK activates the immediately-early oncogenes(c-fos, c-jun), which might play an important role in carcinogenesis of breast cancer and be an early event of oncogenesis of breast cancer. PMID- 12536604 TI - [Role of functional inactivation of p53 from MDM2 overexpression in hepatocarcinogenesis]. AB - OBJECTIVE: This paper was to evaluate the role of p53 mutation, and p53 and MDM2 proteins expression in hepatocarcinogenesis. METHODS: Using streptavidin peroxidase conjugation method (SP), the expression of p53 and MDM2 proteins was observed in 61 cases of primary hepatocarcinomas (HCC) and 59 cases of corresponding paracancerous tissue, among which p53 mutations in exons 5-8 were detected in 21 cases by polymerase chain reaction single-strand confirmation polymorphism analysis (PCR-SSCP). RESULTS: Positive nuclear p53 and MDM2 immunostainings were demonstrated in 57.38% (35/61) and 26.23% (16/61) of HCC, and 1.69% (1/59) and 3.39% (2/59) of corresponding paracancerous tissue, respectively. The expressions of p53 and MDM2 proteins in HCCs were significantly higher than those in paracancerous tissues (P < 0.01). The expressions of p53 and MDM2 were not significantly correlated (P > 0.05). There were 42.86% (9/21) mutations in exon 7 of p53 gene and no mutation was observed in exons 5, 6, 8 in HCCs and in paracancerous tissues. In cases of p53 mutations, there were 66.67% (6/9) of p53 overexpression and 11.11% (1/9) of overexpression of both p53 and MDM2. MDM2 overexpression also appeared in 25% (3/12) of cases without mutation. CONCLUSIONS: Mutation of p53 gene and functional inactivation of p53 resulting from MDM2 overexpression play an important role in carcinogenesis of HCC. It is possible that p53 mutations and MDM2 overexpression induced by other mechanisms are involved in carcinogenesis of HCC. PMID- 12536605 TI - [Effects of Stat3 phosphorylation and expression of c-fos and c-jun proteins on hepatocarcinogenesis]. AB - Expression of phosphorylated Stat3 (p-Stat3), c-fos and c-jun proteins was detected by immunohistochemical technique in 55 hepatocellular carcinomas (HCC) and their surrounding liver tissues. The results showed that the positive rates and signal intensity of p-Stat3, c-fos and c-jun protein in HCCs were significantly higher than those in pericarcinomatous tissues. The expressive intensity of c-fos and c-jun proteins was positively related to p-Stat3 expression in HCCs, whereas the positive correlation of expressive intensity was only observed between c-jun protein and p-Stat3 in pericarcinomatous tissues. The data suggest that the phosphorylation of Stat3 may be an early event in hepatocarcinogenesis; the overexpression of p-Stat3 protein, which activates c fos and c-jun genes, may contribute to malignant transformation of hepatocytes; hepatocytes which expressed p-Stat3, c-fos or c-jun proteins may be potentially malignant pre-cancer cells in pericarcinomatous liver tissues. PMID- 12536606 TI - [Ultrastructural observation and analysis of thin basement membrane nephropathy]. AB - In order to improve the diagnosis of thin basement membrane nephropathy, we observed the ultrastructure of renal biopsy specimens from five cases with thin basement membrane nephropathy, and selectively measured the thickness of the basement membrane. The result showed: 1. The thickness of the basement membrane of five patients was less than 250 nm. 2. The thickness of the basement membrane was positively related to the course of disease (r = 0.65), While negatively related to the grade of haematuria (r = -0.39). The results suggest that hypoplasia may be the cause of extensively thinning of the basement membrane. PMID- 12536607 TI - [Study on molecular epidemiology of HCMV infection in mothers and their newborns in Changsha]. AB - The detection of HCMV-IgG, IgM in plasma by ELISA and DNA in plasma and P(C) BMCs from mothers and their newborns by PCR was carried out. Positive HCMV-IgG, HCMV IgM, P(C)BMCs HCMV DNA and plasma HCMV DNA were demonstrated in 119(95.2%), 7(5.6%), 27(21.6%) and 14(11.2%) in 125 mothers and 117(93.6%), 1(0.8%), 18(14.4%) and 7(5.6%) in their mewborns, respectively. There were significant differences among the positive rate of newborn's HCMV-IgM, plasma HCMV DNA and CBMCs HCMV DNA (P < 0.05), but the difference between HCMV-IgM and plasma HCMV DNA had no statistic significance (P > 0.05). This study suggests that HCMV-IgG and CBMCs HCMV PCR are preferable epidemiological index of HCMV infection of pregnant women and newborns respectively. PMID- 12536608 TI - [Effects of nao-yi-an on expressions of nerve growth factor and interleukin-1 beta following injury induced by hemoglobin in cultured hippocampal neurons]. AB - OBJECTIVE: To investigate the effects of Nao-yi-an (NYA) on protecting the cultured hippocampal neurons against the injury induced by hemoglobin (Hb) and modulating the expressions of nerve growth factor (NGF) and interleukin-1 beta (IL-1 beta) following Hb injury. METHODS: The experimental techniques of neuronal culture, alive cells count, Northern blotting, and enzymelinked immunosorbent assay (ELISA) were employed to detect the survival rate of cultured neurons, the expression level of NGF and IL-1 beta following Hb injury in cultured hippocampal neurons. RESULTS: 1. The hippocampal neurons survived for more than 5 months in Dulbecco's modified Eagle medium (DMEM) containing 2% bovine serum. 2. 38.5% cultured neurons were dead in Lock' s solution with 5 mmol.L-1 glucose containing 50 mumol.L-1 Hb. The rate of survival neurons was significantly increased by addition of decoction of NYA. 3. The expression of IL-1 beta was markedly increased at 1 h, 3 h, 6 h, and 12 h after Hb injury. The expression of NGF was temporarily increased. 4. The levels of IL-1 beta mRNA and protein were significantly decreased in NYA treated group, while the expression of NGF mRNA and protein was not reduced. CONCLUSION: NYA has the effects on increasing the neuronal survival, decreasing the expression of IL-1 beta, and maintaining the expression of NGF following Hb injury in cultured neurons. PMID- 12536609 TI - [Analysis of corneal pathologic changes and laboratory parameters in herpes simplex karatitis patients with ganhuo-shangyanzheng or ganshengyinxuzheng]. AB - In order to investigate the relationship between Ganhuo shangyanzheng and Can shengyin xuzheng in herpes simplex karatitis patients, we observed corneal pathologic changes and examined blood levels of prostaglandin F2 (PGF2), prostaglandin E2 (PGF2), tumor necrosis factor (TNF), arginine vasopressin (AVP), norepinephrine (NR), epinephrine (E) in sixty herpes simplex karatitis patients with Ganhou shangyanzheng or Gan-shengyinxuzheng. The results showed that the corneal pathologic changes were corneal ulcer infiltrating to stroma of cornea in Ganhuo shangyanzheng patients, and refractary corneal ulcer with large amount of corneal neovascularizaton and infiltration of corneal stroma in Gan shengyinxuzheng patients, the blood levels of PGF2, PGE2, TNF, AVP, NE, E in Ganhuo shangyanzheng patients were higher than those in Gan-shengyinxuzheng patients or healthy persons. The results suggest that these parameters may be objective parameters for differential diagnosis of traditional Chinese medicine Zheng types in patients with herpes simplex karatitis. PMID- 12536610 TI - [Effects of di-zhen granules on catecholamine transmitter and gonadotropin of clamacteric rats with deficiency of yin brings about the interior heat-syndrome]. AB - Fifteen-month-old Sprague-Dawley rats were given with some kinds of hot property of herb to produce model of Deficiency of yin brings about the interior heat syndrome. The weight, water drinking volume, and body temperature of model rats were observed at the process of producing model and the treatment of di-zhen granules. Hypothalamic NE, DA and plasma NE, E concentration were determined by high-performance liquid chromatography, and serum E2, FSH, LH levels were determined by radioimmunoassay. The results were that after having been given the hot property of herbs for 2 weeks, the model rats increased water drinking volume and body temperature, which is similar to the appearance of patients with Deficiency of yin brings about the interior heat syndrome, and after having been treated with di-zhen granules, the water drinking volume and temperature of model rats decreased significantly (P < 0.01). The hypothalamic NE, DA, plasma NE, E, and the serum FSH, LH of the group treated with di-zhen granules were significantly lower than that of the model group and the normal group(P < 0.01). The results show that our model mentioned above possesses some characteristic features of the human climacteric syndrome which exhibits Deficiency of yin brings about the interior heat-syndrome. Di-zhen granules may regulate the function of GnRH neurons and other neurotic nucleus in hypothalamus, and then rectify the endocrine disorders of climacteric. PMID- 12536611 TI - [Variability of female peak bone mineral density at different skeletal regions and its effect on diagnosis of osteoporosis]. AB - OBJECTIVE: To evaluate the changes in variability (V) of female peak bone mineral density (PBMD) at different skeletal regions and its effects on diagnosis of osteoporosis. METHODS: Using a Hologic QDR 4500A fan beam X-ray bone densitometer to measure the BMD at different skeletal regions (anteroposterior and supine lateral spine, hip, and forearm) in 1,810 women between the ages of 15-96 from Changsha city (Hunan province) in the People's Republic of China, the population grouped in every 5 years of age, then to get PBMD and its variability. RESULTS: According to every 5 years of age, the analysis showed that PBMD occurred varied in the age of 20-44, the variability ranged 6.0% to 17.0%, in an average of (11.4 +/- 3.1)%, the maximal value (17%) of V is at Ward's triangle, the minimal value of V is at distal one-third site of forearm (6%). The incidence of osteoporosis changed with V of PBMD at different skeletal regions. With different diagnostic standards, the incidences of OP also differed. CONCLUSION: Changes in PBMD and its V at different skeletal regions and the different diagnostic standard obviously affects the incidence of OP. PMID- 12536612 TI - [Changes of ultrastructure and function of the aortic endothelium in streptozotocin-diabetic rats and effect of perindopril]. AB - To investigate the alteration of ultrastructure and function of the aortic endothelium in streptozotocin (STZ)-diabetic rats and the effect of perindopril, male SD rats were randomly divided into normal group (NC), diabetes control group (DC), diabetes group treated with perindopril (2 mg.kg-1.d-1) which was administered after 4 weeks. At 4, 8, 16 weeks after injecting STZ, glucose, plasma endothelin-1 and angiotensin II were respectively measured, and we also observed aortic endothelial cell under the electron microscope. RESULTS: In DC group, there were mitochondrial edema and vacuolization obviously in aortic endothelial cells at 8 weeks, and extensive endothelial cell necrosis and exfoliation were observed at 16 weeks, while pathological changes in the DP group were abated significantly. Plasma Ang II levels were increased significantly at different times in DC group, and plasma ET-1 levels were obviously increased at 8 weeks and greatly decreased at 16 weeks. In DP groups, plasma Ang II levels obviously decreased, ET-1 levels declined at 8 weeks and statistically elevated at 16 weeks. CONCLUSION: There are changes of ultrastructure and function in aortic endothelial cell in different durations of diabetes. Plasma ET-1 level may be a marker of aortic endothelial cell injury, perindopril may have protective effect on aortic endothelial cell in diabetic rats. PMID- 12536613 TI - [Retrospective analysis of argon laser for treating retinal periphlebitis]. AB - OBJECTIVES: This article was to investigate the curative effect of argon laser photocoagulation for retinal periphlebitis treatment, and then to discuss the stage of the treatment. METHODS: According to the condition of vitreous hemorrhage and the range of retinal neovascularization, 31 cases (34 eyes) were divided into the light group and the heavy group, and the therapeutic effective rate, the condition of visual acuity and vitreous hemorrhage were compared. RESULTS: The therapeutic effective rate was 64.71%, there was significant difference in effective rate and prognosis of visual acuity between two groups (P < 0.01). After laser treatment, the morbidity of vitreous hemorrhage in heavy group decreased from 100% to 18.70%. CONCLUSION: Argon laser photocoagulation is an effective method for treating retinal periphlebitis and early treatment is recommended. PMID- 12536614 TI - [Application of blood cells separator in the treatment of high leukocyte acute leukemia]. PMID- 12536615 TI - [High efficiency gene transfer into breast cancer cells by pseudotyped retroviral vector]. AB - OBJECTIVE: To investigate the efficiency of gene transfer mediated by pseudotyped retroviral vector in breast cancer, and to provide an useful vector for breast cancer gene therapy. METHODS: We used a MuLV vector pseudotyped with the vesicular stomatitis virus glycoprotein(VSV-G) envelope to transduce breast cancer cells MDA-MB-435, To compare the transfer efficiency with MuLV. RESULTS: The transfer efficiency is (92 +/- 12)% by using MuLV/VSV-G vector, (24 +/- 5)% by MuLV vector. The transfer efficiency of MuLV/VSV-G is 3.8 times that of MuLV vector. CONCLUSION: Our results suggest that pseudotyped retroviral vector MuLV/VSV-G is a kind of high efficiency gene transfer vector in breast cancer cells. PMID- 12536617 TI - [Radiofrequency catheter abaltion of right atrioventricular accessory pathway by superior vena cava approach]. AB - The radiofrequency ablation effect was observed in 21 patients with single right atrioventricular accessory pathway by superior vena cava approach. Because of many factors, radiofrequency ablation by routine inferior vena cava approach failed. However, all of these accessory pathways were successfully ablated by the superior vena cava approach with could significantly reduce operation and radiofrequency ablation time and exposed time in x-ray, and there were not any complications. The success was probably related to the good contact between the ablation catheter tip and ablation target point. It is suggested that radiofrequency ablation of right atrioventricular accessory pathways by superior vena cava approach is effective and safe when the routine inferior vena cava approach is difficult or unsuccessful. PMID- 12536616 TI - [Clinical study of ischemic preconditioning on isolated lung perfusion with chemotherapeutic agents in the treatment of unresectable lung cancer]. AB - OBJECTIVE: To determine the effect of ischemic preconditioning on isolated lung perfusion (ILP) with chemotherapeutic agents in the treatment of unresectable lung cancer. METHODS: Eight patients with unresectable cancer or metastatic sarcomas in lungs underwent isolated single lung perfusion with doxorubicin. Eight patients were randomly divided into two groups: control group (Group C) and ischemic preconditioning group(Group IP). Group C was only performed isolated lung perfusion with doxorubicin; Group IP was performed isolated lung perfusion with doxorubicin after ischemic preconditioning (in ischemic preconditioning procedure, right or left pulmonary artery was clamped for 10 minutes, then released for 15 minutes). RESULTS: The mean pulmonary artery pressure (MpaP) after ILP in Group IP was much lower than that in Group C (P < 0.05). The PaO2 after ILP in Group IP was much higher than that in Group C(P < 0.01). The lung histologic examination after ILP showed that pulmonary edema, inflammatory cell infiltration, mild focal hemorrhage and alveolar disruption in Group C were significantly serious than those in Group IP, but there was no hospital death in Group C or in Group IP. The complications included hypovolemia shock and acute lung injury. Following up 2 months to 10 months, no death was observed, and the tumours diminished in various degrees or disappeared in the two groups. CONCLUSION: Isolated lung perfusion with chemotherapy can be done safely and effectively in patients with unresectable lung malignancies and metastatic sarcoma in the lung, and ILP can cause lung injury, but lung ischemic preconditioning can reduce the lung injury after isolated lung perfusion. PMID- 12536618 TI - [Level change of IL-10, IL-12 and TNF alpha in the serun of patients with hepatocellular carcinoma]. AB - OBJECTIVE: To study the relationship and clinical significance between level change of serum Interleukin 10 (IL-10), interleukin 12 (IL-12), tumor necrosis factor-alpha (TNF alpha) and primary hepatic carcinoma (PHC). METHODS: Serum IL 10, IL-12, TNF alpha in 40 patients with PHC, 36 cases with hepatocirrhosis and 33 normal controls were measured with ELISA. RESULTS: The higher serum levels of IL-10, IL-12, TNF alpha were observed in patients with hepatocirrhosis than in controls (P < 0.01), although IL-12, TNF alpha in the serum of patients with PHC were in the proximity of that of controls (P > 0.05), the IL-10 level was significantly higher than that of control (P < 0.01), further more, IL-10 being opposed to IL-12, TNF alpha is higher in patients with tumor above 5.0 cm in diameter than below 5.0 cm, IL-12 and TNF alpha were declined and IL-10 increased with tumor enlargement. CONCLUSION: The immune dysfunction in patients with PHC may be related to level increase of IL-10. The level decrease of IL-12, TNF alpha and increase of IL-10 is helpful to clinical differential diagnoses between cirrhotic nodule and PHC, and this suggests cancerous tendency. PMID- 12536619 TI - [Evaluation on severity of esophageal varices by color Doppler parameters]. AB - To assess the value of "left gastric vein shunt index" (LGVSI) in the esophageal varices using color Doppler sonography, 32 variceal patients who had large varices(large varices group), 20 patients who had small or medium varices (small or medium varices group), and 15 control subjects were examined with color Doppler sonography. The results showed that the LGVSI in the large varices group was significantly higher than that in the small or medium varices group and in control subjects (P < 0.01). When the best cut-off value of the LGVSI was 0.12, its sensitivity and specificity were 90.6%, 90.0% respectively. Our results suggest that the LGVSI is a useful Doppler ultrasound parameter in evaluating severity of esophageal varices. PMID- 12536620 TI - [Classification of urogenital tract infection in 518 cases and drug sensitivity analysis]. AB - In 518 cases with urogenital tract infection detected with mycoplasma identification susceptibility testing reagent box, 308 cases (59.4%) were infected by ureaplasma urealyticum (Uu), 21 cases (4.1%) were infected by mycoplasma hominis (Mh), and 189 cases (36.5%) were infected by both Uu and Mh. Drug sensitivity to the macrolides (erythromycin, roxithromycin, josamycin, azithromycin), the tetracyclines (doxycycline, minocin), and the quinolines (ofloxacin, ciprofloxacin) were detected at the same time. The results showed that the sensitivity to various antibiotics in Uu infection was different from that in Mh infection. Macrolides were more sensitive for Uu infection, and quinolines were more sensitive for Mh infection, while the mixed infection of Mh and Uu had resistance to most antibiotics. PMID- 12536621 TI - [Clinical analysis of 15 cases with tetramethylenedisulfotetramine]. AB - In our study, we observed the clinical manifestations of 15 tetramethylenedisulfotetramine (TEM) toxicosis emergency patients diagnosed by plasma toxin chemical analysis, and evaluated the effect of emergency treatments. All cases could not be differentiated from the other kinds of toxicosis by clinical manifestations, and toxin chemical analysis was the sole definite diagnosis method. The effect of gastric lavage in early stage was especially obvious, 13 cases with convulsions got immediate relief after plasma exchange, the use of large dosage of tranquilizer resulted in better control on convulsion in 4 cases treated with ventilator than that in the others. Gastric lavage and chemical toxin analysis in early stage are of vital importance, plasma exchange, tracheotomy and mechanical ventilation are the most effective reatment methods. PMID- 12536622 TI - [Prolactin in normal pregnancy and severe pregnancy-induced hypertension]. AB - OBJECTIVE: This study was to investigate the relationship between serum prolactin and pregnancy-induced hypertension. METHOD: Maternal serum prolactin was estimated by radioimmunoassay in 89 normal pregnant women and 52 patients with severe pregnancy-induced hypertension at 28 to 42 week gestation. RESULTS: In normal pregnancy, prolactin level increased progressively from a mean value of 270 mg.L-1 in the 24th week to 440 mg.L-1 in the 34th week and decreased progressively from the 38th week to the 40th week. In severe pregnancy-induced hypertension increased progressively too; 31 of 52 patients with severe prolactin showed high prolactin levels in zone A (> mean value + standard of prolactin values in the normal pregnancy), 10 patients in zone B (mean + standard to mean) and 11 patients in zone C (< mean). CONCLUSION: These results suggest that high prolactin levels may play an important role in the pathogenesis of severe pregnancy-induced hypertension. PMID- 12536623 TI - [Value of autobiopsy in the diagnosis of peritoneum disease guided by ultrasound]. PMID- 12536624 TI - [Study on diagnosis of small intestinal hemorrhage]. AB - To explore the etiology and different methods of diagnosis for 33 cases of small intestinal hemorrhage, which were confirmed by surgery and/or pathology. METHODS: 33 cases of small intestinal hemorrhage were diagnosed by endoscopy, barium studies, radionucleotide scanning, angiograpy, exploratory laparotomy and intraoperative colonscopy, which were confirmed by surgery and/or pathology. RESULTS: Small intestinal tumors were found in 13 cases (39.39%), Leiomyoma in 8; infectious diseases in 8(24.24%); Meckel's diverticulum in 7(21.21%); and vascular malformation in 3(9.1%) and others in 2(6.06%). Among them, black stool (42.42%) is the most commonly seen. The common clinical presentation was middle anemia and patients with unexplained abdominal pain (36.36%). Radionucleotide scanning was positive in 5 out of 7 cases, and all of five were Meckel's diverticulum. Endoscopy revealed 5 out of 20 cases. Exploratory laparotomy was performed in 6, 3 were found to have some lesions, and the other 3 were diagnosed by intraoperative colonscopy. CONCLUSIONS: Tumor is the nost common cause of small intestinal hemorrhage, mied anemia and black stool are the common clinical presentations, Radionucleotide scanning is one of the most useful diagnostic methods for Meckel's diverticulum and intraoperative colonscopy could elevate veracity of diagnosis. PMID- 12536625 TI - [Delayed cardioprotection of captopril in patients undergoing open heart surgery]. AB - OBJECTIVE: To investigate whether there is delayed cardioprotection of captopril pretreatment in open heart surgery. METHODS: Twenty patients with ventricular septal defect (VSD) undergone open heart surgery were randomly divided into captopril group (CAP group) and control group. In the CAP group, patients were pretreated with captopril (1 mg.kg-1, oral) at 48 hours before operation, No special treatment was given at the same time in the control group. Release of creatine phosphokinase-MB (CK-MB) and myocardial adenosine triphosphate (ATP) content was studied. Heart shock protein 70 (HSP70) in myocardum was examined using Western blotting analysis. RESULTS: No statistically significant difference was found in release of CK-MB between control group and CAP group. ATP depletion was (1.40 +/- 0.56) mumol.g-1, (2.06 +/- 0.72) mumol.g-1 in CAP group and control group respectively (P < 0.05). Western blotting analysis showed that both group had HSP70 expression. In CAP group, HSP70 expression was higher than that in control group. CONCLUSION: The results suggest that captopril pretreatment 48 hours before operation could reduce ATP depletion in pediatric patients of VSD during open-heart surgery, and HSP70 might be involved in the delayed cardioprotection induced by captopril. PMID- 12536626 TI - [Investigation of relationship between concentrated platelets and basic platelets in donors]. AB - OBJECTIVE: To investigate the relationship between the concentrated platelets and basic platelets of donors. METHODS: We observed the harvest rate of concentrated platelets of different basic platelets in donors from Changsha area and non Changsha area. RESULTS: The number of concentrated platelets in Group B was higher than that in Group A (P < 0.01). The harvest rates were 65.4% and 67.5% respectively in donors from Changsha area and non-Changsha area. CONCLUSION: There is positiver relationship between the harvest rate of concentrated platelets and basic platelets, and the transportation of blood does not effect the harvest rate of platelets. PMID- 12536627 TI - [Protective effects of sevoflurane pretreatment on myocardial ischemia reperfusion injury in rabbits]. AB - To observe the myocardial protective effects of sevoflurane pretreatment on ischemia-reperfusion injury, forty eight rabbits were divided into six groups randomly: control group, ischemia-preconditioning group, sevoflurane preconditioning group, ischemia preconditioning plus glyburide group, sevoflurane preconditioning plus glyburide group, glyburide group, each group subjected to three hours of left anterion descending coronary artery occlusion followed by three hours of reperfusion. Infarct size and area at risk were defined by staining. Infarct size [expressed as a percentage of the area at risk (IS:AR)] was (60.8 +/- 10.8)% in controls, and reduced to (33.1 +/- 4.9)% and (30.9 +/- 6.8)% respectively in ischemic preconditioning groups and sevoflurane preconditioning groups. Glyburide pretreatment eliminated the preconditioning effects of ischemia and sevoflurane [IS:AR = (59.3 +/- 11.2)% and (56.6 +/- 11.9)%, respectively; not significant]. Glyburide alone did not increase infarct size [IS:AR = (63.2 +/- 12.5)%, not significant]. It suggests that sevoflurane pretreatment protects the rabbit heart subjected to three hours of coronary occlusion from infarction. PMID- 12536628 TI - [Expression of bcl-2, c-myc genes and their relationship in patients with acute leukemia]. AB - OBJECTIVE: This paper was to explore the expressions of bcl-2, c-myc genes and the relationship between them in bone marrow from patients with acute leukemia (AL). METHODS: In situ hybridization was used to detect bcl-2 and c-myc mRNA. Immune cytochemistry was used to detect bcl-2 and c-myc protein. RESULTS: The expressions of bcl-2, c-myc mRNA and proteins were significantly increased in AL; There were not significant differences in bcl-2 mRNA and protein, c-myc mRNA and protein between the patients with acute lymphocytic leukemia and acute nonlymphocytic leukemia; There were linear correlations between bcl-2 mRNA and bcl-2 protein, between c-myc mRNA and c-myc protein, between bcl-2 mRNA and c-myc mRNA, between bcl-2 protein and c-myc protein. CONCLUSION: bcl-2 and c-myc genes play an important role in pathogenesis and the development of AL. PMID- 12536629 TI - [Clinical diagnosis of lacrimal duct obstruction by digital subtraction dacryocystography]. AB - OBJECTIVE: To illustrate the value of digital subtraction dacryocystography in the diagnosis of lacrimal duct obstruction. METHODS: Eighty-two normal eyes and 56 eyes with epiphora were investigated with digital subtraction dacryocystography and the dacryocystogram were studied. RESULTS: Lacrimal drainage system imaging by the digital subtraction dacryocystography provided reliable and accurate demonstration of lacrimal duct obstruction. CONCLUSION: Digital subtraction dacryocystography is a rapid, effective and noninvasive imaging technique, and it is helpful in the diagnosis, treatment and study of lacrimal duct obstruction. PMID- 12536630 TI - [Using L-form bacterium ATP bioluminescence assay for rapid testing L-form bacterial susceptibility]. AB - Adenosine triphosphate is a kind of necessary metabolites in living cells. The authors detected ATP contents by using bioluminescence method in 111 strains of L form bacteria after exposing to 5 kinds of antibiotics. The results showed that the mean value of CPM was less than (35 +/- 10.2)%.s-1. Thus, the value could be acted as a critical concentration between susceptibility and resistance. The conincidence rate of this method and K-B method was 95.3%. It indicates that the bioluminescence method has a high sensitivity. It can be used to detect L-form bacterium-drug susceptibility quickly and may play an important role for choosing the antibiotics. PMID- 12536631 TI - [Comparison of anesthesia in infant undergoing abdominal surgery]. PMID- 12536632 TI - [Clinical study of oral mucosal diseases caused by the unhealthy prosthesis]. PMID- 12536633 TI - [Preliminary study of the gene structure of human glycosylphosphatidylinositol specific phospholipase D]. AB - To explore the cDNA and its genomic gene structure of human glycosylphosphatidylinositol specific phospholipase D (GPI-PLD), the stromal cells from human bone marrow were cultured, and the GPI-PLD cDNA was successfully cloned from stromal cells (GenBank Accession AY007546). After analyzing this cDNA, we found that it codes the signal peptide as well as the mature peptide (817 AA) of GPI-PLD. Compared with the cDNA cloned from human pancreas and liver, the homology of the cDNA is 99% and 95% respectively. After searching GenBank, we aligned the genomic gene of the GPI-PLD by DNATools software, and found that the GPI-PLD genomic gene was located in the human 6p22.1-22.3 and contained 25 exons, TATA box, CAAT box, enhancer and the sequence binding homo domain of Pit-1/GHF-1. PMID- 12536634 TI - [Infection of mutated mouse complement receptor type II by Epstein-Barr virus]. AB - Site-directed mutagenesis method was used to introduce two desired mutations, which were confirmed by DNA sequencing, into mouse complement receptor Type II gene(MCR2). Then the constructed eukaryotic expression vectors containing wild type mouse CR2/1(wtMCR2/1), mutant type mouse CR2/1 (mtMCR2/1) and human CR2 (hCR2) cDNA were transferred into mouse SP2/0 cells by electroporation. After two week screening by G418, the stably transfected clones were obtained. Several ways including PCR, RT-PCR, and immunohistochemistry were utilized to screen those clones with interesting genes integrated and expressed. Then Epstein-Barr virus(EBV) was used to infect these transfected cells and EBER-1 (EBV encoded RNAs) hybridization results showed that only hCR2 and mtMCR2 transfected SP2/0 cells could be infected by EBV, but positive rate of the former was much higher than the latter. This study sets groundwork for elucidating the mechanism by which EBV enters the cells and for establishing the animal model of EBV-related nasopharyngeal carcinoma (NPC). PMID- 12536635 TI - [Study on base insertion/deletion of CAG-rich region in insulin receptor substrate-1 5'-regulatory sequence]. AB - To study the relationship of the polymorphism of the insulin receptor substrate-1 (IRS-1) gene 5'-flanking regulatory sequence and Type 2 diabetes, the IRS-1 gene 5'-flanking regulatory sequence was scanned by PCR-SSCP in 78 healthy control subjects and 76 Type 2 diabetic subjects. Applying PCR-denatured polyacrylamide gel electrophoresis and silver staining, the insertion/deletion polymorphism of the CAG-rich region was analyzed. The genome DNA of the normal and variant subjects was amplified with high-fidelity pfu DNA polymerase. The purified and digested target fragments were then subcloned into the pCAT Basic vector. Each allele was identified according to the mobility by the restrictive endonuclease digestion of the recombinant combined with denatured polyacrylamide gel electrophoresis and silver staining, and finally the constructive plasmids containing different alleles were analyzed by DNA sequencing. Firstly, we found several insertion/deletion variations in the CAG-rich region of IRS-1 gene. Secondly, 7 genotypes and 6 alleles(T1-T6) in this site were detected. Moreover, T5 and T6 were only observed in Type 2 diabetic group. PMID- 12536636 TI - [Differentiation and gene expression of human osteosarcoma cell line MG-63]. AB - To observe the differentiation and gene expression of human osteosarcoma cell line MG-63 in culture. Alkaline phosphatase (ALP) activity was determined by p nitrophenyl phosphate assay; bone Gla protein (BGP) was measured by radioimmunoassay; type I collagen, matrix metalloproteinase (MMP)-1, tissue inhibitor of matrix metalloproteinase (TIMP)-1 mRNA were examined using semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis; MG-63 cells were stained by the Van GieSon method. Type I collagen mRNA expression achieved a maximum level on the 17th day in MG-63 cells; MMP-1 mRNA was not expressed until the 5th day of culture, and gradually increased; TIMP-1 mRNA was nearly constant; ALP activity gradually up-regulated during 0-12 days, and decreased on the 18th day. By Van GieSon staining, MG-63 cells displayed nodule formation at the 12th day, and became more prominent on the 18th day. The results indicate that human osteosarcoma cell line MG-63 has the osteoblast phenotype; during the differentiation of MG-63 cells, there are the following three principle periods: proliferation, extracellular matrix maturation and mineralization. PMID- 12536637 TI - [Comprehensive assessment of the ovariectomized rat model of postmenopausal osteoporosis]. AB - Dual-energy X-ray absorptiometry(DEXA), scanning electron microscopy, and bone biomechanical test were used to assess comprehensively bone quantity and quality of ovariectomized rats. In OVX rats, not only bone mineral density (BMD) of lumbar vertebrae in vivo and vitro, but also BMD of femora (except for R3 region) and proximal metaphysis(R1 region) in vitro decreased obviously (P < 0.01), whose bone loss rates of L5 and L6 were the highest and achieved 13%; the trabeculae of OVX rats were few, fine, and discontinued and there were lacunae on the surface; in OVX rats, both compressive strength of vertebral bodies and the mechanical properties of femora decreased; the falling degree of the former was greater; the maximal compressive power of lumbar vertebrae decreased with 33.32%. We conclude that after 4-month post-ovariectomy, the bone quantity and quality of six-month old rats decreased, especially in the area of abundant cancellous bone such as vertebral bodies, distal femora; the assessment of the efficiency of new drugs to prevent and treat postmenopausal osteoporosis using rat models should include these drugs' effects to the bone mass, the bone structure, and the bone strength. PMID- 12536638 TI - [Construction of recombinant antisense RNA IgG2aVH region of murine systemic lupus erythematous]. AB - OBJECTIVE: To construct recombinant plasmid of IgG2aVH region antisense RNA of systemic lupus erythematous. METHODS: Total RNA was isolated from spleen cell of BWF1 mice. Using spleen cell total RNA as the template, We designed specific primers from A6.1 region sequences, and amplified the IgG2aVH region 375 bp DNA fragments by RT-PCR. The IgG2aVH cDNA was cloned by T/A and inserted into pcDNA 3.1 plasmid of vector. RESULTS: The IgG2aVH antisense RNA plasmid expressing recombinant was identified by restriction enzyme digestion and DNA sequence analysis. CONCLUSION: There is IgG2aVH gene in F1 mice spleen cell; the IgG2aVH antisense RNA expressing recombinants is constructed successfully. PMID- 12536639 TI - [Effects of lorsartan, fosinopril on myocardial fibrosis, angiotensin II and cardiac remolding in hypertensive rats]. AB - OBJECTIVE: To investigate effects of lorsartan, fosinopril on myocardial fibrosis, angiotensin II and cardiac remolding in the spontaneously hypertensive rats (SHR). METHODS: 16-week-old SHRs were divided randomly into 3 groups: SHR-L (treated with lorsartan), SHR-F (treated with fosinopril) and SHR-C (untreated), each group consisting of 10 rats. After 8 weeks' and 16 weeks' therapeutic period, collagen volume fraction (CVF), perivascular circumferential area (PVCA), plasma and myocardium angiotensin II concentrations were examined by pathological examination with computed processing and radioimmunoassay respectively. RESULTS: (1) Compared with SHR-C after 8 weeks' and 16 weeks' therapeutic period, the systolic blood pressure (SBP) was decreased similarly in both treatment groups. Heart and left ventricular weights, heart weight and eft ventricular mass indexes were lower significantly in both treatment groups than in SHR-C. Left ventricular mass index was reduced to a lower extent in SHR-F group than in SHR-L group after 16 weeks. (2) Compared with SHR-C, CVF, PVCA after 8 weeks and 16 weeks were reduced significantly in SHR-F and SHR-L. Meanwhile, CVF after 16 weeks in SHR-F than in SHR-L. (3) Compared with SHR-C after both therapeutic periods, plasma and myocardium angiotensin II concentrations were increased Significantly in SHR-L, but plasma angiotensin II concentrations were not altered significantly in SHR-F. However, myocardium angiotensin II concentrations were reduced significantly in SHR-F after 8 weeks and 16 weeks in SHR-F. CONCLUSION: Lorsartan, fosinopril inhibit myocardial fibrosis and reverse heart hypertrophy. Fosinopril may be more effective in these above effects than Lorsartan. The mechanism of the both drug's cardioprotective effects was related to inhibition of myocardium rennin angiotension-aldsteron system. PMID- 12536640 TI - [Study on p53 gene expression following transient ischemia in the gerbil]. AB - OBJECTIVE: To study the expression of p53 gene after transient ischemia in the gerbil. METHODS: Northern blot was used to detect the expression of the p53 mRNA in the forebrain during different periods after post-ischemic reperfusion in the gerbil. RESULTS: The expression of p53 mRNA increased in the forebrain during the 6th hour to the 3rd day of reperfusion following 6 min of ischemia (P < 0.05). CONCLUSION: The high level expression of p53 might induce neuronal apoptosis after ischemia. PMID- 12536641 TI - [Study on expression of PCNA and estrogen, progesterone receptors in endometrial carcinoma]. AB - OBJECTIVE: To investigate the proliferating ability of tumor cells in 74 cases with endometrial carcinoma (EMC), as well as define the correlation between clinical pathology, prognostic features and estrogen receptor (ER), progesterone receptor (PR). METHODS: Proliferating cell nucleus antigen (PCNA) and ER/PR content were examined using immunohistochemical assay. RESULTS: PCNA was expressed in both benign hyperplasia and EMC, but positive rate in the latter was significantly higher than that in the former (82.4% vs 30.0%) (P < 0.01). EMC well differentiated had lower proliferating index (PI) score than those moderately and poorly differentiated(P < 0.001), and the PI score was negatively related to the grade of differentiation(r = -0.52, P < 0.01). PI score in early cases (I/II) was significantly lower than that in advanced ones(III/IV) (P < 0.05), and the PI score was positively related to the clinical stages (r = 0.55, P < 0.01). Positive rates of ER, PR were 66.2% (49/74) and 75.7%(56/74) respectively, expression of PCNA was negatively related with ER and PR(r = -0.42, -0.51, respectively, P < 0.05). CONCLUSIONS: EMC cells with poorer differentiation or in more advanced stages have faster proliferating ability and aggressive biological behavior. Expression of PCNA combined with detection of ER/PR not only apts to retrospective study, but also has some prognostic value. PMID- 12536642 TI - [Study on the differentiation and apoptosis of HL-60 cell line induced by Puerarin]. AB - OBJECTIVE: To investigate the differentiation of HL-60 cells induced by different doses of Purerarin(PR) comparing with all-trans retinoic acid(ATRA) and to see if PR can induce apoptosis of HL-60 cells. METHODS: Cell differentiation was analyzed by NBT reduction, the ratio of NBT/MTT and CD11b, apoptosis by morphology, DNA electrophoresis, and flow cytometry(FCM). RESULTS: 80 micrograms.ml-1, 160 micrograms.ml-1, 320 micrograms.ml-1 PR could induce differentiation of HL-60 cells, no significant difference was observed between the cells treated with 1 mumol.L-1 ATRA and 320 micrograms.ml-1 PR. Treated with 320 micrograms.ml-1, 640 micrograms.ml-1 PR, HL-60 cells exhibited a morphological characteristic of apoptosis and typical DNA ladder on gel electrophoresis. FCM analysis showed that PR could interfere with cell cycle in HL-60 cells, with a increased ratio of sub-G1 in HL-60 cells. CONCLUSION: PR exerts effect on differentiation and induces apoptosis in HL-60 cells. PMID- 12536643 TI - [Inhibitory effect of the combination of DHPAs and hyperbaric oxygen on Japanese B encephalitis virus infection in mice]. AB - OBJECTIVE: To investigate the inhibitory effect of the combination of 2,3 dihydroxypropyl-adenine (DHPAs) and hyperbaric oxygen (HBO) on Japanese B encephalitis virus infection in mice. METHODS: Mice infected P3-55 and P3-53 virus were used in the experiments. Virus was inoculated intraperitoneally to Kumming strain mice (8.5-9.5 g). Mice were divided into groups and treated in the incubation period of the infection. The duration of treatment was 5 days, and the experiments were terminated at the end of 2 weeks after treatment. The living and the dead were then counted finally. RESULTS: The intergroup comparison showed marked difference between the control group and combination of DHPAs and hyperbaric oxygen group (P < 0.025 and P < 0.01), but there was no statistical difference between other groups and the control group (P > 0.05). CONCLUSION: The combined treatment of (S) DHPA and/or (RS)-DHPA and HBO has inhibitory effect on Japanese encephalitis virus infection in experimental mice. PMID- 12536644 TI - [Effects of diclofenac sodium combined with dionine in cases with fibrinous membrane after intraocular lens implantation]. AB - OBJECTIVE: The effects of diclofenac sodium(DFNa) combined with dionine in cases with fibrin exudation membrane on intraocular lens (IOL) were studied. METHODS: Thirty-two eyes, derived from sixteen adult pure bred New Zealand rabbits, were divided at random into two groups after extracapsular lens extraction with posterior chamber IOL implantation: (1) rabbits received DFNa eyedrops combined with dionine eyedrops; (2) rabbits received Pred forte eyedrops. The sum of fibrinous exudation membrane on IOL was observed on 1, 3, 5, 7, 14, 21, 30d postoperation. RESULTS: The fibrin exudations in the DFNa combined with dionine group is less than the pred forte group (P > 0.05). CONCLUSION: DFNa combined with dionine is effective in treating fibrin exudation membrane after extracapsular lens extraction and IOL implantation, and it is more effective than the pred forte. PMID- 12536645 TI - [Assessment of dry body-weight in hemodialysis patients by plasma cyclic 3'5' guanosine monophosphate]. AB - OBJECTIVE: The aim of this paper was to investigate the changes in cyclic 3'5' guanosine monophosphate (cGMP) before and after hemodialysis to estimate the value of cGMP to the dry boby-weight. METHODS: Plasma cGMP levels (by radioimmunoassay), cardiothoracic ratio (CTR), and the body weight (BW) before and after hemodialysis were determined in chronic hemodialysis patients and clinical signs and symptoms were observed at the same time. RESULTS: 1. The predialytic cGMP value of the patients was significantly higher than that of healthy controls (P < 0.05). 2. The postdialytic cGMP level was significantly lower than the predialytic cGMP level (P < 0.01). 3. Postdialytic CTR and BW values were significantly lower than predialytic values (P < 0.01). 4. Compared to those of predialysis, postdialytic clinic signs and symptoms of the patients were significantly relieved. CONCLUSIONS: 1. The plasma cGMP level can sensitively reflect the hydration state and is a reliable marker for dry body weight estimation. 2. The measurement of plasma cGMP combined with clinical parameters and radiological indexes permit a more accurate dry body-weight estimation. PMID- 12536646 TI - [Clinical use of interlocking intramedullary nail treating in complex and nonunion fracture of femur and tibia]. AB - Fifty-one complex and nonunion fractures of femur and tibia in forty eight patients were treated with interlocking intramedullary nail. All patients were followed-up for 6 to 29 months to evaluate the curative effect. Forty-nine fractures in forty-six patients were healed(95.8%). The time of healing was 16 weeks(10-24 weeks) in femur and 18 weeks (12-28 week) in tibia in fresh fracture, and 20 weeks(14-46 weeks) in femur nonunion and 18 weeks (12-28 weeks) in tibia nonunion. In femur, the efficacy of 28 cases was excellent or good (93.3%), and 2 cases mediate (6.7%); in tibia, 18 cases were all assessed as excellent or good. The complications were observed in 8 patients (16.67%), including distal screw misinserting, extremity shortening, screw loosening, distal shaft fracture and fracture nonunion. The results suggest that interlocking intramedullary nail can be used in many kinds of femur and tibia fracture with satisfactory result, especially in the complex fracture and nonunion after fixation failure. PMID- 12536647 TI - [Study on in vitro nitric oxide and interleukin-6 levels induced from peripheral mononuclear cells in patients with osteosarcoma]. AB - Peripheral blood mononuclear cells from 32 cases of osteosarcoma and 20 normal controls were separated and induced by lipopolysaccharide, followed by 48 hour incubation in vitro, then the supernatant were collected. The levels of IL-6 were measured by enzyme linked immunosorbent assays, the concentrations of NO were measured by Griess methods. The results were as follows: The concentrations of IL 6 and NO were significantly higher than those in normal controls (P < 0.01). There were positive correlation between the levels of IL-6 and NO in patients with osteosarcoma (r = 0.652, P < 0.01). The results suggest that the immune function of peripheral mononuclear cell in patients with osteosarcoma was disordered. It may activate peripheral mononuclear cells to produce high levels of IL-6 and NO, which may take part in the pathogenesis of osteosarcoma. PMID- 12536648 TI - [A measure of chronic respiratory disease questionnaire for clinical trail]. AB - OBJECTIVE: The aim of this paper was to investigate the reliability and validity of the Chronic respiratory disease questionnaire (CRQ) in China. METHOD: Sixty eight patients with chronic respiratory disease who were in hospital from January to November, 1999 were surveyed with CRQ. Eighteen patients were investigated repeatedly with CRQ in one week to test the reliability; before and the 10th day after treatment, 50 patients were surveyed twice with CRQ and the peakflow of these patients were also detected. The same doctor explained questionnaires and tested peakflow. RESULTS: The correlated analysis of reliability was positive (r = 0.732, P < 0.01). The correlated analysis between the difference of twice questionnaire scores and the difference of twice-peakflow value was also positive (r = 0.565, P < 0.01). CONCLUSION: CRQ can be used by the clinical doctors of China. PMID- 12536649 TI - [Study on endothelial dysfunction and platelet activation in elderly hypertensive patients complicated with cerebral infarction]. AB - OBJECTIVE: The aim of this paper was to investigate endothelial dysfunction, platelet activation, and inflammation in elderly hypertensive patients and those complicated with cerebral infarction. METHODS: Twenty-eight elderly hypertensive patients complicated with cerebral infarction(within 72 hours after onset of neurological symptoms), thirty-one Stage I-II elderly hypertensive patients and twenty-eight elderly healthy individuals were selected as subjects. Their plasma concentrations of von Willebrand factor(vWF), alpha granule membrane protein 140(GMP-140), and serum concentrations of C-reaction protein(CRP) were examined. RESULTS: Elderly hypertensive patients complicated with cerebral infarction had significantly higher plasma vWF, GMP-140, and serum CRP than elderly hypertensive patients and elderly healthy individuals. Plasma vWF and GMP-140 were higher in elderly hypertensive patients than elderly healthy individuals, whereas serum CRP was increased slightly and there was no significant difference between elderly hypertensive patients and elderly healthy individuals. CONCLUSION: This study suggests that elderly hypertensive patients complicated with cerebral infarction have obvious endothelial dysfunction, platelet activation, and inflammatory change. Stage I-II elderly hypertensive patients have obvious endothelial dysfunction and platelet activation as well. PMID- 12536650 TI - [Relationship between sex hormone levels and blood calcitonin gene-related peptide/endothelin-1 in postmenopausal women with coronary heart disease]. AB - OBJECTIVE: To investigate the relationship between sex hormone levels and blood calcitonin gene-related peptide/endothelin-1, and to evaluate the therapeutic effects of nylestriol. METHODS: Forty perimenopausal women without coronary heart disease (CHD) and 30 postmenopausal women with CHD were studied, and 30 postmenopausal women were divided into two groups and treated randomly with either 2 mg nylestriol or placebo. Serum 17 beta-estradiol (E2), testosterone (T), plasma calcitonin gene-related peptide (CGRP) and endothelin-1 (ET-1) were measured. RESULTS: In postmenopausal women, serum levels of E2 and plasma concentrations of CGRP decreased, while serum levels of T and plasma concentrations of ET-1 increased; E2 was positively related to CGRP, and negatively related to ET-1; T was negatively related to CGRP or ET-1. In the nylestriol group, compared with the parameters before treatment, plasma CGRP increased and ET-1 decreased after treatment. CONCLUSION: Estradiol possesses certain effects on endothelial cell, which may be an important aspect of its protective effects. PMID- 12536651 TI - [Surgical treatment of 219 cases of primary tumor and cyst of the mediastinum]. AB - Two hundred and nineteen patients with primary tumor and cyst in the mediastinum were retrospectively analysed. All patients received surgical treatment and pathologic examination. Results showed that: (1) the commonest mediastinal tumors was the thymoma (42.0%) (2) The second teratoma (20.1%). The incidence of neurogenic tumors was 16.4%. The over-all incidence of these three tumors was 78.5%. A total excision of the tumor was performed in 198 patients; incomplete excision was performed in 18 patients; 3 patients underwent biopsy only. The anatomic locations, clinical manifestations, complication and mortality of surgical treatment of these tumors were analysed. PMID- 12536652 TI - [Analysis of monoclonal protein in 72 cases of multiple myeloma]. AB - Serum and urinary monoclonal proteins (M protein) were measured in 72 cases of multiple myeloma (MM) using rate nephelometry. In IgG and IgA types of MM, the level of immunoglobulin (Ig) corresponding to the malignant isotype was significantly higher and that of Ig uncorresponding to the malignant isotype lower than the normal level. The light chain corresponding to the malignant isotype in serum was increased and the light chain uncorresponding to that in serum was decreased either kappa-IgG, IgA types or lambda-IgG, IgA types. Either kappa-LC or lambda-LC type of MM, the serum light chain corresponding to the malignant isotype was in the normal range and uncorresponding to that was decreased, and the corresponding light chain in urine was significantly elevated. Kappa/lambda ratio in serum and urine was all significantly abnormal in IgG, IgA, and LC types of MM. Our data suggest that any quota among kappa light chain > 20 g.L-1 or < 5 g.L-1, lambda light chain > 10 g.L-1 or < 2 g.L-1 in serum and kappa/lambda ratio > 5 or < 0.75 in serum and urine has an important value for diagnosing MM. PMID- 12536653 TI - [Surgical treatment of congenital heart defect complicating severe pulmonary infection in infants and young children]. AB - OBJECTIVE: To review the experience of surgical management of left to right shunt congenital heart defect complicating severe pulmonary infection. METHOD: Twenty patients aged 4-17 months who weighed 3.6-11.6 kg were analysed. Among them, 16 underwent surgical treatment. RESULTS: The mortality rate of 4 who received conservative therapy was 75% (3/4). The mortality rate of the operative group 16 Cases, was 12. 5% (2/16), 14 were doing well by follow-up of 8-24 months. CONCLUSION: Infants of Congenital heart defect Complicating Severe Pulmonary infection, Should be performed operation actively. PMID- 12536654 TI - [Clinical efficacy of burning mouth syndrome treated by livial]. AB - OBJECTIVE: To observe the clinical efficacy of livial on postmenopause women with burning mouth syndrome. METHODS: Fifty six postmenopause women with burning mouth syndrome were randomly divided into two groups, 26 patients were treated with livial as the treatment group, 30 patients were treated with oryzanol and vitamin E as the control. To evaluate the effect, all the patients were observed in 3-6 months after therapy. RESULTS: The result showed that the total effective rate of the treatment group was 84.62% after 3 months, 88.46% after 6 months, and significantly higher than that in the control(P < 0.005). CONCLUSION: It is indicated that livial is safer and more effective than nylestriol in treatment with burning mouth syndrome. PMID- 12536655 TI - [Clinical features of acute myocardial infarction in the young and middle-aged patients: analysis of 68 cases of coronary artery angiography]. AB - OBJECTIVE: This study was to evaluate the clinical features of acute myocardial infarction (AMI) in young and middle-aged patients. METHODS: Eighty-seven patients were classified into two groups according to the age. Among them, 61 patients were 60 years old and above, 26 patients below 50 year old. Selective coronary artery angiography was performed on 68 cases and a scoring method which reflects the extent of narrowing of coronary artery was also used. RESULTS: The older group had more risk factors than the young group. The angina pectoris in the young group was less than the older group. The incidence of acute heart failure in the young group was higher than the older group. The mortality of both two groups had no significant differences. CONCLUSION: The high risk of AMI in the young and middle-aged patients should be paid more attention in the clinic. PMID- 12536656 TI - [Relationship between vertical root fracture and the habits of chewing betel nut]. AB - OBJECTIVE: The aim of this article was to study the causes of vertical root fractures. METHODS: Age and gender, personal history of the sickness and clinical features of vertical root fracture in 27 patients were studied. RESULTS: Vertical root fracture tended to occur in patients with a higher mean age (54 years) and were more frequent in male (70%) than in female. Mesial or mesial-buccal roots of the first molars (78%) were the predilection site of vertical root fracture. All of the patients had chewed betel nut for 10 to 25 years. The occlusal surfaces were abrasive. CONCLUSION: Vertical root fracture is associated with an excessive, repetitive, and heavy masticatory stress applied to a tooth. PMID- 12536657 TI - [Evaluation of clinical therapeutic efficacy by injecting adriamycin to the nerve in primary trigeminal neuralgia]. AB - Sixty-eight cases suffering from primary trigeminal neuralgia were performed by injecting adriamycin to the nerve. The results show that the symptoms of trigeminal neuralgia completely disappeared in all the cases during two weeks after the operation in which 63.2% in 24 hours, 28.9% in one week and 7.9% in two weeks. the recurrence rate in three years was 10.3% and in three to five years was 15.4%. This technique has the features of low recurrence rate, steady therapeutic effect, safety and microtrauma. PMID- 12536658 TI - [Spiral CT evaluation in diagnosis of endobronchial tuberculosis]. AB - OBJECTIVE: To determine the value of spiral CT in diagnosis of endobronchial tuberculosis (EBTB) after analysing the spiral CT appearance. METHODS: Retrospective analysis of CT signs in 13 cases who were finally pathologically found suffering from EBTB via biopsy with bronchoscope. RESULTS: Five cases have signs of bronchial structure with obstructive atelectasis and pneumonia. Two cases have signs of irregular bronchial wall. Four cases have signs of scattering dot and patchy increase density in lung fields. Two cases' bronchi and lung tissues are normal. CONCLUSION: CT examination can determine the site and extent of EBTB and help the localization of bronchoscope. PMID- 12536659 TI - [Telomerase activity in bladder cancer tissues]. AB - OBJECTIVE: To study the correlation between telomerase and bladder cancer. METHODS: The telomerase activity was detected by telomerase PCR ELISA in 41 cases of bladder cancer tissues and 23 cases of non-cancerous bladder tissues. RESULTS: The telomerase activity was positive in 34 cases of bladder cancer tissues (82.9%) and 1 case of non-cancerous tissues (4.3%); the difference of positive rate was significant (P < 0.01). CONCLUSIONS: Telomerase activity is related with bladder cancer closely; telomerase activation may be an important molecular step in the initiation and development of bladder cancer; telomerase may be an excellent molecular biomarker in finding bladder cancer at an early stage and in postoperative following-up. PMID- 12536660 TI - [Study of Chlamydia trachomatis infection on cervical secretion of women with early pregnancy and secondary sterility]. AB - OBJECTIVE: This study was to investigate the Chlamydia trachomatis (CT) infection rate in cervical secretion of early pregnant and secondary sterility women. METHODS: CT-DNA in the cervical secretion of the early pregnant, the secondary sterility, and the control women was detected with polymerase chain reaction (PCR). RESULTS: The infection rates of CT in the early pregnant and secondary sterility women (14.28% and 13.33%) were higher than in the control women (3.33%, P < 0.05), and there was no significant difference between the former two groups (P > 0.05). CONCLUSION: There are higher infection rates in the early pregnant and secondary sterility women. Detecting CT and treating in the perinatal period are needed in the early pregnant women who want to be pregnant again. PMID- 12536661 TI - [Effects of percutaneous transluminal coronary angioplasty and stenting on QT dispersion in patients with coronary heart disease]. AB - OBJECTIVE: The aim of this study was to approach the effects of percutaneous transluminal coronary angioplasty (PTCA) and stenting on QT dispersion (QTd) in patients with coronary heart disease. METHODS: PTCA and stenting were performed successfully on 42 patients with coronary heart disease. QTd and corrected QTd (QTcd) were obtained with a standard 12-lead ECG before and after PTCA + Stent. RESULTS: QTd and QTcd after PTCA + Stent were reduced significantly compared to those before PTCA + Stent (P < 0.01). There were no significant difference in QTd and QTcd before PTCA + Stent between single vessel lesion and multi-vessel lesion, but after PTCA + Stent, QTd and QTcd in single vessel lesion were decreased significantly compared to those in multi-vessel lesion. The ventricular arrhythmia in 9 patients was over after PTCA + Stent. CONCLUSIONS: QTd and QTcd were decreased significantly after PTCA + Stent because of the improvement of myocardial ischemia and heterogeneous repolarization in patients with coronary heart disease. The degree in decreasing QTd and QTcd was associated with compensatory circulation in coronary artery. PMID- 12536662 TI - [A comparative study of serum sFas in patients with hepatocellular cancer and chronic hepatitis]. AB - OBJECTIVE: To study the effects of sFas in hepatocellular cancer (HCC) and chronic hepatitis (CH). METHODS: The serum sFas was detected in 18 patients with HCC, 12 patients with CH and 6 cases of normal control by ELISA. RESULTS: The serum sFas in HCC was obviously increased and had significant difference with the patients of CH and normal control (P < 0.01). The serum sFas had positive correlation with the serum TBIL(P < 0.01), but negative correlation with the ALB, PTA and the ratio of ALT/AST(P < 0.01). CONCLUSIONS: sFas may resist the occurrence of HCC apoptosis. In CH, sFas has correlation with the severity of CH. The role of sFas in viral hepatitis is uncertain. PMID- 12536663 TI - [A case of the recurrent left atrial mignant tibrous histocytome]. PMID- 12536664 TI - [Urinary RBP and glucocorticoid hormone therapy sensibility in adult primary nephrotic syndrome]. AB - OBJECTIVE: To clarify the relationship between the renal tubular function and the efficacy of glucocorticoid, in adult patients with primary nephrotic syndrome. METHODS: Pro- and post-therapy urinary RBP and NAG were determined double antibodies sandwich ELISA and color comparimetry with p-nitrate reductase, respectively, in eighty adult patients with primary nephritic syndrome, according to the concentrations of urinary protein, these patients were divided into three no remission group, partial remission group and fully remission group, fifty-one normal persons as control group. Renal tubular function parameters among groups were compared before and after therapy, and the results were analysed when those parameters were used to predict the sensibility of glucocorticoid therapy. RESULTS: 1. There were no significant differences in urinary RBP, NAG and protein levels before treatment among patient groups (P > 0.05); 2. There were significant decreases in urinary RBP and NAG following glucocorticoid therapy in those response to glucocorticoid (P < 0.01); 3. There were significant differences in urinary protein, RBP and NAG among three groups of patients with primary nephrotic syndrome after treatment. CONCLUSION: Determination of urinary RBP may predict the sensibility of the therapy in the adult patients with primary nephrotic syndrome and its diagnostic efficiency is better than urinary NAG. PMID- 12536665 TI - [Isolation of high-quality genomic DNA from plants]. AB - In order to isolate high-quality genomic DNA from medicinal plant tissues enriching polyphenols and polysaccharides, a simple and rapid method based on CTAB extraction for isolating high-quality intact DNA was established by modifying several existing methods. With this technique, the absorbance ratio (A260/A280) of DNAs obtained from fresh and/or dried roots of Panax ginseng, P. Quinquefolius and P. notoginseng was 1.8 approximately. The restriction fragments of DNAs were directly digested with restriction enzyme (EcoR I/Mse I), linked up by T4 DNA ligase and amplified by nested PCR. Reproducible amplified fragment length polymorphism (AFLP) genomic DNA fingerprinting profiles were established with the isolated DNAs. The results demonstrate that the modified technique may be efficient and reliable in isolating high-quality and high-molecular-weight DNAs from fresh and/or dried medicinal plants containing a high content of polyphenols and polysaccharides. We expect that this method can also be applied to other plants. PMID- 12536666 TI - [Establishment of two-dimensional gel electrophoretic protein map of human spermatozoa]. AB - OBJECTIVE: To establish the method of 2-dimensional electrophoresis(2-DE) for proteins of human spermatozoa and to construct a protein map of human spermatozoa. METHODS: The sperm pellet was prepared with simple Percoll layer protocol. We studied the effects of various sample preparation methods, loading quantities and isoelectric-focusing protocols on the quality of silver-stained 2 DE map, and constructed a primary protein map of human spermatozoa. RESULT: Up to 703 protein spots were acquired with sample preparation Method I while only 194 210 spots with Method II. With immobilized pH gradients and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(IPG-DALT) we could acquire over 700 spots while only 280-300 with isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis(ISO-DALT). CONCLUSION: It is satisfactory to lyse sperm with sample preparation Method I and to separate sperm proteins by IPG DALT for establishing 2-D map of human sperm. PMID- 12536667 TI - [Establishment of method for determining serum beta-hydroxybutyric acid and its meaning in diabetes ketosis acidism]. AB - OBJECTIVE: This study was to establish enzyme-rate method for determining serum beta-hydroxybutyric acid, and to discuss its meaning in diabetes ketosis acidism. METHODS: Enzyme-rate method was used to determine serum beta-hydroxybutyric acid in 60 cases of normals, 30 cases of diabetes, and 8 cases of ketosis acidism by Hitachi 7170A autochemistry analyzer. RESULTS: The recovery rate is 108%, the linerity extent 0-4.0 mmol.L-1, the coefficient of variation within-run is 2.2% and day-to-day is 2.6%. CONCLUSION: The enzyme-rate method for determining serum beta-hydroxybutyric acid save time, gives accurate results and has wide linerity, so it can used for diagnosis of diabetes ketosis acidism. PMID- 12536668 TI - [Clinical study on 143 cases of myelodysplastic syndrome]. PMID- 12536669 TI - [Treatment of deep infection after total knee joint replacement]. PMID- 12536670 TI - Caspase inhibitors as anti-inflammatory and antiapoptotic agents. AB - The striking efficacy of Z-VAD-fmk in the various animal models presented above may reflect its ability to inhibit multiple enzymes including caspases. In accord with this, more selective, reversible inhibitors usually show low efficacy in multifactorial models such as ischaemia, but may offer some protection against NMDA-induced excitotoxicity and hepatitis. Importantly, caspase inhibitors may exhibit significant activity in vivo even when they are applied post insult. As far as the CNS is concerned, the first systemically active inhibitors have emerged. Functional recovery could be achieved in some ischaemia models, but long term protection by caspase inhibitors is still being questioned. Recent developments in drug design enabled the first caspase inhibitors to enter the clinic. Although initially directed towards peripheral indications such as rheumatoid arthritis, caspase inhibitors will no doubt eventually be used to target CNS disorders. For this purpose the peptidic character of current inhibitors will have to be further reduced. Small molecule, nonpeptidic caspase inhibitors, which have appeared recently, indicate that this goal can be accomplished. Unfortunately, many fundamental questions still remain to be addressed. In particular, the necessary spectrum of inhibitory activity required to achieve the desired effect needs to be determined. There is also a safety aspect associated with prolonged administration. Therefore, the next therapeutic areas for broader-range caspase inhibitors are likely to involve acute treatment. Recent results with synergistic effects between MK-801 and caspase inhibitors in ischaemia suggest that caspase inhibitors may need to be used in conjunction with other drugs. It can be expected that, in the near future, research on caspases and their inhibitors will remain a rapidly developing area of biology and medicinal chemistry. More time, however, may be needed for the first caspase inhibitors to appear on the market. PMID- 12536671 TI - RNA as a drug target. AB - In the antiviral and antibacterial area, increasing drug resistance means that there is an ever growing need for novel approaches towards structures and mechanisms which avoid the current problems. The huge increase in high resolution structural data is set to make a dramatic impact on targeting RNA as a drug target. The examples of the RNA binding antibiotics, particularly, the totally synthetic oxazolidinones, should help persuade the skceptics that clinically useful, selective drugs can be obtained from targeting RNA directly. PMID- 12536672 TI - ACAT inhibitors: the search for a novel and effective treatment of hypercholesterolemia and atherosclerosis. PMID- 12536673 TI - Growth hormone secretagogues: discovery of small orally active molecules by peptidomimetic strategies. PMID- 12536675 TI - [Influence of human cytomegalovirus infection on the expressions of HOXB1, HOXB5, HOXB6, and HOXB9 genes in human embryo lung cells]. AB - OBJECTIVE: To study the expressions of HOXB1, HOXB5, HOXB6, and HOXB9 genes in human embryo lung (HEL) cells and influence of human cytomegalovirus (HCMV) infection on the expressions of these genes. METHOD: The expressions of HOXB1, HOXB5, HOXB6, and HOXB9 genes were detected with semi-quantitative RT-PCR method. RESULTS: 1. HEL cells expressed HOXB5 and HOXB6 genes, but they did not express HOXB1 and HOXB9 genes. 2. After HCMV infection, HEL cell was induced to express HOXB9 gene, as the expression of HOXB6 gene was elevated. The expression of HOXB5 gene had no significant change. HOXB1 was still not expressed. 3. Treated with all-trans retinoic acid (ATRA), the expression of HOXB9 gene in HEL cells infected by HCMV was significantly increased. But during the advanced infect period, the expression of HOXB6 was significantly decreased. CONCLUSION: HCMV can induce abnormal expressions of HOXB6 and HOXB9 genes, which may play an important role in HCMV-induced abnormal embryogenesis. PMID- 12536674 TI - Inhibitors of hepatitis C virus NS3.4A protease: an overdue line of therapy. PMID- 12536676 TI - [Effect of hepatitis C virus NS3 protein on expression of iNOS mRNA in hepatocarcinogenesis]. AB - OBJECTIVE: To investigate hepatocarcinogenesis by detecting the effect of HCV NS3 protein on expression of inducible nitric oxide synthase (iNOS) mRNA in hepatocellular carcinoma (HCC) and pericarcinomatous liver tissues(PCLT). METHODS: The expression of HCV NS3 protein and iNOSmRNA was detected by immunohistochemical technique (SP method) and in situ hybridization in specimens of HCC and PCLT from 52 patients with HBV(-). The relationship between the positive rate and the signal strength was tested statistically. RESULTS: The positive rate of HCV NS3 protein in HCC was lower (46.2% vs. 67.3%), and signal strength was weaker than those in PCLT. The signal strength of HCV NS3 protein was correlated with the degree of carcinomatous cells differentiation in HCC (P < 0.01). The positive rates (80.8% and 90.4%) of iNOSmRNA in HCC and PCLT had no statistical difference. Signal intensity of iNOSmRNA in HCC was weaker than that in PCLT. The higher HCVNS3 protein expression level was, the stronger iNOSmRNA signals in PCLT (P < 0.01). In particular, the localization and patterns of HCV NS3 protein were much similar to those of iNOSmRNA in the PCLT of some patients. CONCLUSIONS: HCV NS3 protein may exert its hepatocarcinogenic effect in the early stage on host cells by increasing product of nitric oxide, which may bring about transformation of hepatocytes. PMID- 12536677 TI - [Studies on mutation of isoniazid resistant genes in M. tuberculosis]. AB - OBJECTIVE: To understand mutation of isoniazid-resistant genes in M. tuberculosis clinical isolates, and to develop the method for rapid detection of drug resistance. METHOD: Analyzing the katG genes in 30 M. tuberculosis clinical isolates with PCR-SSCP(single stranded conformation polymorphism) and PCR-DS (direct sequencing) techinques. RESULTS: With the reference of M. tuberculosis strain H37 Rv was used as control, all isoniazid-sensitive isolates and 4 isoniazid-resistant isolates displayed ab normal PCR-SSCP and PCR-DS patterns. But of 12 isoniazid-resistant isolates, 4(33.4%) isolates displayed abnormal katG PCR-SSCP pattern, 8 (46.7%) isolates displayed abnormal katG PCR-SSCP pattern, 5(41.7%) isolates were missense mutation at codon 94, 3(25%) isolates were cosense mutation at codon 96. CONCLUSIONS: Resistance to isoniazid in most M. tuberculosis isolates is due to the mutations on katG genes. PCR-SSCP techniques could be used as a method for rapid detection of isoniazid-resistance in M. tuberculosis. PMID- 12536678 TI - [Changes in somatosensory evoked potentials and brainstem auditory evoked potentials during acute intracranial hypertension in rabbits]. AB - OBJECTIVE: To study the changes in somatosensory evoked potentials (SEPs) and brainstem auditory evoked potentials (BAEPs) during intracranial hypertension. METHOD: A rabbit model of acute intracranial hypertension was developed. Thirty New Zealand rabbits were divided into 3 group. Both Group A and B were groups with intracranial hypertension; Group C was the control group. In Group A, balloons were placed to the right parietal lobe beneath the dura mater; in Group B, to temporal at the middle fossa near the brainstem. RESULT: (1) In Group A, when ICP rose to (1.48 +/- 0.35) kPa, P1 amplitude of SEPs was significantly changed, whereas latencies of Wave IV, V, and VII of BAEPs were prolonged when ICP rose to (9.40 +/- 1.60) kPa. In group B, P1 amplitude of SEPs was changed when ICP reached (8.10 +/- 1.25) kPa, and latency of Wave III of BAEPs was prolonged when ICP rose to (3.11 +/- 0.80) kPa. (2) P1 amplitude increased with ICP value when the balloon volume increased from V = 0.0 ml, ICP = (0.43 +/- 0.10) kPa to V = 0.4 ml, ICP = (2.44 +/- 0.65) kPa and then decreased afterward. CONCLUSIONS: (1) Changes in SEPs and BAEPs relate to not only ICP value but also the location of the balloon. (2) With the increasing of ICP, P1 amplitude increases firstly and then decreases. PMID- 12536679 TI - [Effects of losartan on renal ultrastructure in diabetic rats]. AB - OBJECTIVE: To observe the effects of losartan on renal ultrastructure in streptozotocin diabetic rats. METHODS: Male SD rats were randomly divided into 2 groups: normal control group (NC group) and diabetic group. Diabetic group was induced by streptozocin (65 mg.kg-1) abdominal injection. Four weeks later, diabetic rats were further divided into 2 groups: diabetic rats treated with losartan (DL group, 20 mg.kg-1.d-1, by gavage) and diabetic unteated control group (DC group). Renal ultrastructure of each group was observed before and after 12 weeks of treatment respectively. RESULTS: The ultrastructure alterations in DL group including mesangial expansion and thickening of glomerular basement membrane (GBM), were lighter than those in DC group. CONCLUSION: Losartan can prevent renal pathological progress in diabetic rats. It is suggested that losartan may have some renal protective effects. PMID- 12536680 TI - [Effect of peritonitis pathomorphology on rabbit model in peritoneal dialysis]. AB - The study about peritonitis rabbit model is rarely, although peritonitis have been considered mainly complication in peritoneal dialysis. Twelve New Zealand rabbit were divided into peritonitis group and control group. The peritonitis group rabbit were injected with the dialysate containing alive staphylococcus aureus and the control ones with 0.9% saline into the peritoneum. The pathomorphology changes of the peritonitis were observed. The results showed (1) There was proliferation and swollen of mesothelial cells, vessel dilation and infiltration of inflammation cells in the peritoneum in the peritonitis group; and no such changes in the control group. (2) The D/D0 glucose concentration in the dialysate of peritonitis group was lower than that of the control ones at any point except the 0 min (P < 0.01). (3) The ratio of Cr concentration in the dialysate to Cr concentration in the plasma in the peritnitis group was significantly higher than that of the control group (P < 0.05). (4) The WBC count in the dialysate of peritonitis group was obviously higher than that of the control group all points except 0. These results implied that after injection of alive staphylococcus aureus to the peritoneal cavity of New Zealand rabbit for 48 hours, the pathomorphology change was similar with the characteristics of peritonitis complicated with peritoneal dialysis in patient. PMID- 12536681 TI - [Expression of metalloproteinases and tissue inhibitor of metalloproteinases in human osteoblastic MG-63 cells and normal human osteoblasts]. PMID- 12536682 TI - [Experimental research of protective and therapeutic effects of zinc and vitamin E on mouse liver radiational damage]. AB - OBJECTIVE: To study the effects of zinc and vitamin E on lipid peroxide (LPO) contents and glutathione peroxidase (GSH-Px) activities in liver homogenates of radiational damage mice. METHODS: Two hundred and forty mice were divided randomly into five groups: normal control group (Group A), 60Co gamma-ray irradiator group (Group B), 60Co + zinc (Zn) group (Group C), 60Co + vitamin E(VE) group (Group D), and 60Co + Zn + VE group (Group E). After irradiated with 60Co gamma-ray 7.5 Gy, the mice were protectively treated with Zn and VE at different times. LPO contents and GSH-Px activities in liver homogenates of the mice were determined. RESULTS: The level of liver LPO was significantly higher (P < 0.01), but the level of liver GSH-Px was significantly lower (P < 0.01) in Group B than those in Group A; the content of liver GSH-Px was markedly higher in Group C and Group D than that in Group B (P < 0.01), while the content of liver LPO was markedly lower in Group D than that in Group B (P < 0.05); the concentration of liver GSH-Px was markedly higher in Group E than that in Group D (P < 0.01). CONCLUSIONS: VE may play an important role in lowering liver LPO contents and raising liver GSH-Px activities in radiational damage mice. vitamin E can cooperate with zinc in raising liver GSH-Px activities, stopping liver cells from lipids peroxidation and keeping integrity of liver cell membrane. PMID- 12536683 TI - [Growth inhibition of epidermal growth factor-stimulated human glioblastoma cells by nicardipine in vito]. AB - OBJECTIVE: To study the growth effects of epidermal Growth Factor (EGF) and the calcium channel antagonist nicardipine (NC) on U251 MG, a human malignant glioma cell lime, which has high-affinity EGF receptors. METHODS: The growth effects of EGF and NC on U251 MG cultured in serum-free and serum-containing (10% fetal bovine serum, FBS) media respectively were observed by MTT colorimeritric analysis. RESULTS: (1) EGF significantly enhanced the growth of U251 MG in a dose dependent manner in serum-free conditions. The near-maximal effect was obtained at 10 mg.ml-1. The addition of serum obscured this effect. (2) NC decreased U251 MG cells proliferation, especially in serum-containing media, and completely blocked the growth-stimulated effect of EGF. The combinated effects of EGF (10 ng.ml-1) and NC equaled those of NC alone. CONCLUSIONS: When serum is absent U251 MG cells showed a pronounced mitogenic response to EGF in a dosedependent manner, which approximated that achieved with 10% FBS. NC suppressed the growth of U251 MG cells and completely blocked the growth-stimulated effects of EGF, it can be used as an adjuvent therapy for human malignant gliomas. PMID- 12536684 TI - [Experimental study on using diclofenac sodium or dionine topically to treat fibrinous membrane after intraocular lens implantation]. AB - OBJECTIVE: To find a reasonable method for treating and preventing fibrinous membrane after intraocular lens (IOL) implantation by using diclofenac sodium or dionine topically. METHODS: Forty-eight eyes of twenty-four adult pure bred New Zealand rabbits were divided into three groups randomly. Group A: rabits received diclofenac sodium eyedrops; Group B: rabbits received dionine eyedrops; Group C: controlled. All the eyes received lens extraction combined with posterior chamber intraocular lens implantation. The fibrinous membrane was observed after 1, 3, 5, 7, 14, 21 and 30 days postoperatively. RESULTS: The fibrinous membran in Group A was less than that of Group B and Group C, (P < 0.05); the difference between group B and group C on fibrinous membrance was also statistically significant (P < 0.05). CONCLUSION: The study suggests that both diclofenac sodium and the dionine eyedrops can treat fibrinous membrane after IOL implantation. The effect of diclofenac sodium is better than that of dionine. PMID- 12536685 TI - [Bcl-2 and bax expression in rat cerebral ischemic preconditioning]. AB - OBJECTIVE: To study the expression of apoptosis regulated genes bcl-2 and bax nocoprotein expression and its significance in rat cerebral ischemic preconditioning. METHODS: With immunohistochemical assay, bcl-2 and bax expression were studied in rat brains which were divided into four groups: the sham-operation control group, the cerebral ischemic group and the cerebral ischemic preconditioning group. RESULTS: Bcl-2 oncoprotein was induced in cerebral ischemic preconditioning control group. Bax oncoprotein expression had no obviously difference in four groups. CONCLUSIONS: The expression of bcl-2 and bax oncoprotein after sublethal ischemic preconditioning might be associated with the preconditioning protection against neuronal damage following subsequent lethal ischemia. PMID- 12536686 TI - [Study on telomerase activity and its clinical value in human non small cell lung cancer]. AB - OBJECTIVE: To investigate telomerase activity and its clinical value in human non small cell lung cancer. METHODS: Using the method of TRAP-PCR-ELISA, we measured telomerase activity in human non small cell lung cancerous, adjacent, lung benign lesion, and normal lung tissues. RESULTS: The positive rate of telomerase activity was 75%(27/36) in lung cancerous tissues; 6.25%(2/32) in adjacent tissues; 11.1%(1/9) in lung benign lesion tissues; and 0%(0/9) in normal lung tissues. There was no significant difference in telomerase activity among the pathological type, the degree of tissue differentiation, the size of tumor, lymph node metastasis, and clinical stage in the patients with lung cancer. CONCLUSION: The telomerase activity may become an important molecular marker in the diagnosis and differential diagnosis of human non small cell lung cancer. PMID- 12536687 TI - [Experimental study of pBMP/MPTCP in bone defects therapy]. AB - To develop a new substitute of bone graft, which is composed of porcine bone morphogenetic protein (pBMP) and the magnetic porous tricalcium phosphate (MPTCP), we investigated the effects of pBMP/MPTCP (Group A), autologous red bone marrow/porus tricalcium phosphate (BM/PTCP) (Group B), pBMP/PTCP(Group C) on the repair of bone defects created surgically in rabbits. By gross inspection, roentgenography examination, histological examination, immunohistochemistry test of osteocalcim, quantitive histological measurement of new bone formation and biomechanical test, three kinds of substitutes were proved to be biocompatible. Group A induced more new bone formation than other two groups, and showed a better biomechanical function(P < 0.01). It is believed that pBMP/MPTCP has characteristics of strong osteoinductive capacity, good biocompatibility, excellent filling of bone defects and gradual biodegradation. PMID- 12536688 TI - [Detection of serum Lp(a) level of coronary heart disease and its clinical significance]. AB - OBJECTIVE: To study the association of human serum Lp(a) level and coronary heart diseases. METHODS: Objects examined were conposed of 2 groups: CHD (39 cases) and healthy controls (52 cases). Lp (a), TC, HDL-C, TG, apoB of two groups were determined and the results were done with statistic analysis. RESULTS: The mean serum Lp(a) concentrations (mg.L-1) in coronary heart disease group were shown higher significantly than that in the control group (P < 0.01). However, there were no significant correlation between the mean serum Lp(a) and the mean serum TC, HDL-C, TG, spoAI and aopB.(P > 0.05). CONCLUSIONS: Serum Lp(a) level is closely related to the occurrence of coronary heart disease. Lp(a) is a single risk factor for coronary heart disease. Detecting the determination of serum Lp(a) is extremely valuable to the clinical prediction and diagnosis of CHD. PMID- 12536689 TI - [Clinical application of a new nasolacrimal stent]. AB - Seventy-two patients(ninety-one eyes) with epiphore due to obstruction of the nasolacrimal duct underwent nasolacrimal stent of our own disign. The period postoperative follow-up was 6 to 20(mean 14.5) months. Seventy-six eyes demonstrated obvious resolution of epiphore, twelve eyes had partial resolution, and three eyes had no resolution. The success rate were 96.7%. The common complications included occluded stent and a slightly blood-stanied nasal discharge. The placement of the nasolacrimal stent is a safe simple and effective method for treatment of epiphore due to obstruction of the nasolacrimal duct. PMID- 12536690 TI - [Changes in plasma concentration of interleukin-6 in elderly patients with coronary artery disease complicated with heart failure]. AB - OBJECTIVE: The aim of this study was to observe the concentrations of plasma interleukin-6(IL-6) in elderly patients with coronary artery disease(CAD). METHODS: Forty-eight CAD patients with heart failure, 15 CAD patients without heart failure, and 30 normal subjects were studied. The plasma concentration of interleukin-6(IL-6) was measured by enzyme-linked immunosorbent assay. RESULTS: The plasma concentration of IL-6 in CAD patients with heart failure was significantly higher than that without heart failure and normal subjects, respectively (P < 0.05). There was no significant difference between CAD patients without heart failure and normal subjects in the plasma levels of IL-6. There was a negative correlation between the concentration of IL-6 and left ventricular ejection fraction(LVEF) (r = -0.60, P < 0.01). The plasma concentration of IL-6 of the group with re-hospitalization because of severe heart failure was significantly higher than that of the group without hospital readmission (P < 0.01). CONCLUSION: The plasma concentration of IL-6 in CAD patients with heart failure may provide the important information for the severity of the disease and for the prognostic prediction in the disease. PMID- 12536691 TI - [Relationship between protein loss and dialysis duration results of PET and KT/Vurea in CAPD patients]. AB - The study focused on the relationship between protein loss and dialysis duration, results of PET and KT/Vurea in two groups of CAPD patients. 16 patients of one group were included in CAPD for only 30 days while 19 patients of the other group were included in CAPD for more than one year. The results showed that value of protein loss had no obvious significance between both groups. Compared with the patients with value of PET D/Pcr < 0.65, the patients with value of PET D/Pcr > or = 0.65 had no significant protein loss. Between the patients whose index of urea clearance > or = 2.0, and those index of uera clearance < 2.0 the protein loss had no difference. It is suggested that, in routine dialysis duration peritoneal transport of small solutes and dialysis adequency. The conclusion shows furtherly that transportion of large protein and small solutes through different ways. PMID- 12536692 TI - [MRI analysis of non-traumatic multiple vertebral collapse in aged people]. AB - OBJECTIVE: To evaluate the MRI in the differential diagnosis of acute non traumatic vertebral collapse. METHODS: Sixty-eight cases were retrospected. All cases were studied with sagittal T1-weighted MR images, T2-weighted MR images, gadolinium-enhanced T1-weighted MR images, and axial T1-weighted MR images. Twelve of 68 were performed extra coronal T1-weighted MR images and 22 with additional axial gadolinium-enhanced T1-weighted images. RESULTS: The causes of vertebral collapse were: (1) osteoporosis (n = 31), their images showed retropulsion of bony fragments of collapsed vertebral body which kept the normal signal into the spinal canal as the distinctive sign; (2) metastatic tumor (n = 30), their images showed destructions of pedicles of vertebral and epidural mass as the distinctive sign, and hyperintense or mixed hyperintense signal on T2WI and enhanced T1WI of compressed vertebrae; and (3) multiple myeloma (n = 7), their images showed long T1WI and long T2WI, without destruction of pedicle and epidural mass in compressed vertebrae. CONCLUSION: MRI is useful to the differentiations of the non-traumatic vertebral collapse in aged. PMID- 12536693 TI - [Clinical study of the effect of buflomedil on peripheral neuropathy in diabetic patients]. AB - OBJECTIVE: To observe the effect of buflomedil on clinical symptom and nerve conduction velocity (NCV) of diabetic peripheral neuropathy patients. METHODS: 58 cases of diabetic patents with peripheral neuropathy were divided into diabetes group treated with buflomedil (DB) 100 mg.d-1 and diabetes control group (DP) treated with PGE1 200 micrograms.d-1. NCV was measured and peripheral nerve symtom was observed in two groups before treatment and after 2 weeks. RESULTS: NCV and peripheral nerve symptom were obviously improved both in DB group and DP group (P < 0.01). The effective rate were respectively 80.0% and 89.3% in DB group and DP group. The diversity was no significant in DB group and DP group (P > 0.05). CONCLUSION: buflomedil is an effective and safe drug for treating diabetic peripheral neuropathy patients. PMID- 12536695 TI - [Statistical analysis of 47 cases with Holt-Qram syndrome]. AB - By statistical analysis of 47 cases with Holt-Qram syndrome(HOS), we found that the severity of the upper limb abnormalities and cardiac defects in HOS varied significantly with different individuals. The variations of appearance were related with the types and positions of mutatons of TBX5 gene which could damage the gene function and cause HOS. It is suggested that the genetic heterogeneity in HOS may be caused by the mutations of different genes. PMID- 12536694 TI - [Analysis of P interval dispersion and its correlative factors among Chinese]. AB - OBJECTIVE: To establish mean value of P interval dispersion of 12-lead simultaneous body surface electrocardiogram in healthy Chinese and to analyse its correlative factors. METHODS: 12-lead simultaneous body surface electrocardiogram data were taken from 2078 healthy Chinese aged from 1 to 87 [mean (39.4 +/- 17.3)] years with SR-1000A automated electrocardiography made by Zhongshan, Guangdong province. These data were stored in the floppy disk. RESULTS: Mean value of Pmax, Pmin, Pd and Pcd seemed to be prolonged, and Pcmax, Pcmin to be shortened with increasing of age, but there was no significant difference between age and sex. Mean value of Pmax of the all subjects (1-87 years) was (109.85 +/- 7.88) ms, Pmin (80.97 +/- 11.42) ms, Pcmax (123.13 +/- 12.00) ms, Pcmin (90.85 +/ 14.40) ms, Pd (28.89 +/- 10.74) ms, Pcd (32.39 +/- 12.10) ms; mean values of Pmax, Pmin, Pd and Pcd among the children of 1-14 years were (101.54 +/- 9.49) ms, (76.85 +/- 10.19) ms, (24.82 +/- 7.51) ms and (31.10 +/- 9.89) ms, respectively; 77.04% of Pmax was in II lead and 14.39% in V5 lead, 74.69% of Pmin was in V1 lead and 7.84% in V2 lead. CONCLUSION: The mean value of Pmax is 96-120 ms, Pd 8-50 ms and Pcd 10-56 ms among healthy Chinese. Pd is not significantly related with heart rate, age and sex. PMID- 12536696 TI - [CT diagnosis of hemorrhagic renal cysts]. AB - CT signs of nineteen cases of hemorrhagic renal cysts that surgically and pathologically proved were analyzed. 8 of the 19 cases revealed as high density hemorrhagic lesion. CT signs in those 8 patients include low density rim sign, cyst immerging sign, and perirenal fascia thickening. The other 11 of 19 cases revealed as low density lesion. CT signs include thickened cyst wall and inhomogeneous density of the cystic content. The hounsfield unit of the lesion is correlated with the duration of disease. It is suggested that CT is valuable in diagnosis of high density hemorrhagic renal cysts, and it gives a clue to the diagnosis of low density hemorrhagic renal cysts. PMID- 12536697 TI - [Influence of zafirlukast on pulmonary functions and quality of life in patients with asthma]. AB - OBJECTIVES: To explore the influence of zafirlukast on pulmonary functions and quality of life (QOL) in patients with mild-to-moderate asthma at the stage of attack and at the stable stage. METHODS: Patients at the stage of attack were randomly divided into two groups: Group one is zafirlukast 20 mg twice daily plus inhaled the half quantity of glucocorticoid(budelade) of the Global Initiative for Asthma (GINA) standard quantity, long-effection aminophyllin 0.2 g twice daily and inhaled salbutamol when necessary; Group two: inhaled budelade of GINA standard quantity, long-effection aminophyllin 0.2 g twice daily and inhaled salbutamol when necessary). Patients' peak expiratory flow (PEF)/PEF predicted value % and delta PEFR% were determined at the second week and the forth week before and after the treatment. On the standard treatment, patients with the stable stage were added zafirlukast 20 mg twice a day. PEF values in the morning and evening and QOL were evaluated. RESULTS: After treating with zafirlukast, the PEF/PEF predicted value % and delta PEFR% in Group one were higher and lower than those of Group two respevtively. These changes were appeared in two days and were peaked in two weeks and the peak had been gone on for more than 4 weeks, but these were no significant difference between the Groups (P > 0.05); PEF values in the morning and evening in patients with the stable asthma increased 5.9% and 4.6% respectively and their QOL were improved in asthma symptoms, limitative activities, avoiding stimuli, and reacting on stimuli. CONCLUSION: Zafirlukast can improve pulmonary function, reduce the inhalation quantity of steroid in patients with mild-to-moderate asthma, and play an important role in treating and improving QOL in patients with the stable asthma. PMID- 12536698 TI - [Study of delayed encephalopathy after acute carbon monoxide poisoning by clinical characteristic, CT and MRI features]. AB - OBJECTIVE: To explore the diagnosis value of CT and MRI in delayed encephalopathy after acute carbon monoxide poisoning (DEACMP). METHODS: The clinical, CT and MRI data of 20 patients with DEACMP were analysed. RESULTS: The main clinical manifestations of the disease were intellectual disturbances and personality disorders. CT scan in DEACMP showed that some characteristic changes (such as diffuse low density) was observed within bilateral cerebral white matter, prominently within bilateral or unilateral globus pallidus or basal ganglia areas. The MRI showed that brain lesions in DEACMP were bilateral symmetric punctate, spotty or confluent foci within periventricular white matter and centrum semioval, which showed high signal intensity in T2-weighted and low signal intensity in T1-weighted. The chronic ischemia changes were found in the globus pallidus or basal ganglia areas. Cerebral cortex lesions were observed occasionally. CONCLUSIONS: The diagnosis of DEACMP mainly depends on case history, clinical features and neuroradiological presence. MRI is more valuable in the diagnosis and the differential diagnosis than CT. PMID- 12536699 TI - [Expression of urokinase-type plasminogen activator and its receptor protein in synovial tissues from osteoarthritis]. AB - OBJECTIVE: To study the expression of urokinase-type plasminogen activator (uPA) and its receptor (uPAR) in the synovial tissures, and the significance of uPA and uPAR in degradation of extracellular matrices in osteoarthritis (OA), then analyse the possible relationship between uPA and uPAR. METHODS: Immunohistochemical analysis technique was used to detect uPA and uPAR protein expression and distribution in synovial tissues in 26 OA patients and 10 mormal individuals. RESULTS: Positive staining of uPA and uPAR protein were detected in 19 cases in the 26 OA samples (73%), while only 2 positive cases were seen in the 10 mormal tissues (20%). The expression rate of uPA protein was markedly higher in OA than in the normal (P < 0.01). The expression of uPAR was seen in 14 cases in all 26 OA samples (54.6%), while there was only one case in 10 normal samples (10%), the positive rate was significantly higher in OA than in normal samples (P < 0.05). Positive expression of uPA and uPAR proteins were found in synovial lining cells, mononuclear cells, macrophage-like cells and endothelial cells. Using the correction analysis, we found a positive correlation between uPA and uPAR reactivity in the synovial tissues in OA (r = 0.920, P < 0.01). CONCLUSIONS: The high expression of uPA and uPAR protein in OA synovial tissues indicate that the uPA system may play an important role in the process of synovitis and degradation of OA cartilage extracellular matrices. In the pathologic process of OA, uPA and uPAR coordinate each other, lead to the genesis and development of OA cartilage degradation. PMID- 12536700 TI - [Effects of noninvasive positive pressure ventilation on gas exchange and patients' transformation in chronic obstructive pulmonary disease and respiratory failure]. AB - OBJECTIVE: This paper was to observe the effects of noninvasive positive pressure ventilation (NPPV) on gas exchange and patients' transformation and to evaluate their clinical value. METHODS: Patients were randomly treated by either NPPV, or conventional medicine. The arterial blood gas readings, heart rates (HR), and respiratory rates (HR) were obtained at admission after the treatment. It was necessary to intubate when the clinical deterioation was marked by PaCO2 > 70 mmHg, PaCO2 gradually increasing 5-10 mmHg, pH decreasing 0.05-0.1 or mental obstacle, or PaO2 < 45 mmHg. RESULTS: NPPV significantly decreased the PaCO2 level, HR, and RR, significantly increased the PaO2 level, and also decreased the opportunity of intubation. CONCLUSION: The results suggest that NPPV can improve the gas exchange and reduce the need of intubation in COPD patients with acute respiratory failure. PMID- 12536702 TI - [A case of embryo reduction of a five gestation pregnancy]. PMID- 12536701 TI - [Relationship between serum thyroid hormone and GH-IGF axis, growth failure in nephrotic rats]. AB - OBJECTIVE: To investigate the individual effects of malnutrition, nephrosis and glucocorticoid therapy on serum thyroid hormone, and explore the relationship between serum thyroid hormone and GH-IGF axis, growth failure in nephrotic rats. METHODS: Twenty-four male Sprague-Dawley (SD) rats were randomly divided into control group, pair-fed group, doxorubincin-induced nephrotic group (nephrotic group) and dexamethasone-treated nephrotic group (des-treated group). Serum T3/T4, GH and IGF-I were measured by RIA, serum IGFBPs were measured by Western ligand blot, liver GHR and IGF-I/IGFBPs mRNA were analyzed by radio-receptor assay and RT-PCR respectively. RESULTS: 1. Serum thyroid hormone was low in pair fed group, lower in nephrotic group and lowest in des-treated group except for high serum T4 in pair-fed group. 2. Serum thyroid hormone was positively related with liver GHR, IGF-I/IGFBP-3 mRNA and serum IGF-I/IGFBP-3, and negatively related with serum IGFBP-2. 3. Serum thyroid hormone was positively related with nose-tail length and weight (both P < 0.01). CONCLUSIONS: The hypothyroidism is a possible mechanism of reducing serum IGF-I levels and biologic action, and resulting in growth failure in nephrotic syndrome. PMID- 12536703 TI - [Analysis of 48 cases of brain abscess]. AB - To analyze brain abscess for its origins of infection, diagnosis, and methods of treatment, 48 patients with brain abscess were retrospectively reviewed and they were treated with nonoperative way, puncture and aspiration, and resection. The results were that the cure rate was 81.25%, the improvement was 12.5%, and the mortality was 6.25%. The chronic otogenic infection and cryptogenic infection are the main origins of infection of brain abscess. The diagnosis of brain abscess depends on CT and MRI, and methods of its treatment are decided by the condition and speciality of brain abscess. PMID- 12536704 TI - [Relationship between the axially lymph nodes metastases and the bone metastases in patients with the breast cancer]. AB - OBJECTIVE: To investigate the relationship between the axially lymph nodes metastases and the bone metastases in patients with the breast cancer. METHODS: Lymphadenotomy of the axially lymph nodes had been performed on patients with the breast cancer, and the excised axially lymph nodes were observed by pathological examination. All patients underwent postoperatively total bone scan with single photon emission computer tomography (SPECT) periodically. RESULTS: Among 292 patients with the breast cancer, the bone metastatic rates of low and high differentiating tumor were 27.3% (6/22) and 45.2% (122/272) respectively, the bone metastatic rates in both tumors had no difference (P > 0.1). The bone metastatic rate was 54.3% (100/184) in patients suffered from the axially lymph nodes metastases, and 25.9% (28/108) in non-metastases patients respectively, the difference of their bone metastatic rates was significant (P < 0.001). The bone metastatic rate was 48.4% (62/128) in patients suffered from the metastases of axially lymph nodes after operation less 2 years, and 67.8% (38/56) after operation more than 2 years respectively, the difference of their bone metastatic rates was not significant (P < 0.5). CONCLUSIONS: There is no relationship between the bone matastatic rates and the pathological type of tumor. The bone metastatic rates of patients with the axially lymph nodes metastases are higher than those with non-metastases of axially lymph nodes. In addition, the bone metastatic rates of patients with the axially lymph nodes metastases increase with time prolongation. PMID- 12536705 TI - [Analysis of early feeding and gastrointestinal dysfunction in neonates with asphyxia]. AB - OBJECTIVE: This article was to investigate the effect of early initiation of enteral feeding in neonates with perinatal asphyxia on gastrointestinal function. METHODS: Eighty-four cases of enteral feeding of neonates with asphyxia were analysed. RESULTS: The common dysfunctions were vomiting and abdominal distension, gastric bleeding as well; the percentages of gastrointestinal dysfunction in the early four days were 65.2%, 24.1%, 8.9%, 1.12% respectively and presented a decreasing tendency following the increasing days after birth; The percentage of gastrointestinal dysfunction on the first day was the highest in the early three days (P < 0.05) and not only dextrose but also dilute milk resulted in the dysfunctions. Fasting caused the lower percentage of the dysfunction(2.15%, P < 0.05), but dextrose and dilute milk had no significant difference(P > 0.05). CONCLUSION: It's not optimal for neonates with asphyxia to initial early enteral feeding, especially in the early three days; it's optimal for the neonates with asphyxia not to be fed on the first day. PMID- 12536706 TI - [Clinical significance and the treatment of myocardial bridge]. AB - To explore the incidence and significance of myocardial bridge in the patients undertaken coronary angiography, 861 patients were retrospectively analyzed. Fourteen myocardial bridges was detected and the prevalence rate was 1.6%. All myocardial bridges were in left anterior descending branches and among which 1 with Grade I stenosis, 4 with Grade II stenosis and 9 with Grade III stenosis. All cases had clinical symptoms and electrocardiographic changes in myocardial ischemia. Eight cases were performed test of exercise and 7 were positive and 1 was suspected positive. Three patients examined nuclide myocardial perfusion showed anterior wall myocardial ischemia of left ventricle. After having been treated by beta-blocking agents, symotoms of angina pectoris were relieved obviously and after 0.5-6 year's follow-up, no acute myocardial infarction, sudden death, and acute heart failure occured in patients with myocardial bridge. The prognosis in patients with myocardial bridge was better than those with coronary heart diseases with stable stenosis. Myocardial bridge in coronary arteries may cause myocardial ischemia and angina, and therapy of beta-blocking agents is an effective treatment. PMID- 12536707 TI - [Treatment of femoral neck fractures in the elderly patients with the compression screw]. AB - The treatment of 50 patients above 70 years with femoral neck fractures were reviewed. All the patients were treated by internal fixation with the compression screw. Followed-up 1-5 years reviews showed that the rate of fracture union was 83%, the satisfactory rate of the function was 96%, no death was found. It is suggested that internal fixation with compression screw may be a good treatment in the elderly patients with femoral neck fractures. PMID- 12536708 TI - [Research of cryosurgery in the treatment of maxillo-facial deepish hemangioma]. AB - OBJECTIVE: To observe the effect of cryosurgery on maxillofacial deepish hemangioma, and the facial nerve recovery. METHODS: Twelve hemangioma patients were observed for the effect of cryosurgery on hemangiomaa in maxillo-facial deepish expose lesion after flapping and four rabbits were used for the observation of the recovery of the facial nerve after cryosurgery. RESULTS: The facial nerve function of rabbits recovered within six to eight weeks, the twelve hemangioma patients'-facial nerve also recovered with six to eight weeks, only one reoccured in five years. CONCLUSIONS: Cryosurgery after flapping is an effective; little-traumatic, recover-quickly and spreadingworth method for treatment of maxillo-facial deepish hemangioma. PMID- 12536709 TI - [Treatment of epistaxis by radiofrequency]. AB - OBJECTIVE: To evaluate the therapeutic effect of radiofrequency on epistaxis. METHOD: Three hundred and ten patients with epistaxis were treated with radiofrequency and Nd:YAG laser, respectively. RESULT: The effective rate of radiofrequency was 88.8%, Nd:YAG laser 90.5%. CONCLUSION: The therapeutic effects of epistaxis by radiofrequency and ND:YAG laser are similar, but radiofrequency is simpleier, easy to perform, more and useful than Nd:YAG laser. PMID- 12536710 TI - [Pattern reversal visual evoked potentials analysis in patients with noninsulin dependent diabetes mellitus]. AB - OBJECTIVE: To explore the alteration of pattern reversal visual evoked potentials (PVEPs) in patients with noninsulin-dependent diabetes mellitus (NIDDM) and its values in clinical application. METHODS: PVEPs were recorded in 30 cases with NIDDM and 30 cases of normal individuals. RESULTS: The abnormal rate of PVEPs in patients with NIDDM was 58.33%, and the latency of P100 was markedly prolonged. The wave-forms were poor-differentiated, and the amplitudes of waves were low. CONCLUSION: Abnormal PVEPs can be regarded as one of the index of visual function in patients with NIDDM. The abnormality of PVEPs is related to blood sugar in empty stomach, course of NIDDM. peripheral nerve lesions and diabetic retinopathy. PMID- 12536711 TI - [Efficacy of prostaglandin E1 in the treatment of chronic pulmonary heart disease in the elderly people at the acute stage]. AB - OBJECTIVE: To investigate the effects of prostaglandin E1 (PGE1) in the treatment of chronic pulmonary heart disease in the elderlys at the acute stage. METHODS: Forty cases of chronic pulmonary heart disease at the acute stage were treated with PGE1 at the dosage of 100 micrograms.d-1 for 7 days by intravenous drop, other 40 patients with the same disease were treated without PGE1, and 20 healthy subjects were selected as controls. The level of plasma D-dimer and serum fibrin degradation products (FDP), and blood PaO2 and PaCO2 were determined. The patients' symptoms and signs were observed before and after the treatment. RESULTS: The levels of plasma D-dimer and serum FDP were significantly increased (P < 0.01) in both the PGE1 group and the control group (no PGE1) compared to the healthy subjects. In PGE1 and no PGE1 groups the levels of plasma D-dimer and serum FDP were significantly decreased (P < 0.01) after the treatment. The level of PaO2 was increased and there was no significant difference (P > 0.05) between the two groups, but the level of PaCO2 was significantly decreased (P < 0.01) in both PGE1 and no PGE1 groups, but the decreasing in the PGE1 group was lower than that in the no PGE1 group. The symptoms and signs in PGE1 group were more markedly improved than those in the no PGE1 group. CONCLUSIONS: PGE1 is effective in the treatment of chronic pulmonary heart disease in old people at the acute stage. PMID- 12536712 TI - [Lithotripsy for residual cholelithes by rigid choledochoscopy]. AB - OBJECTIVE: To evaluate a new lithotomy for postoperatively residual cholelithes. METHOD: Using rigid choledochoscopy to extract the residual cholelithes which could hardly be extracted by fibrocholedochoscopy through the T-tube sinus in 31 cases. RESULT: The residual cholelithes were extracted in 30 cases, reoperation was avoided. CONCLUSION: The residual cholelithes can be effectively extracted by the rigid choledochoscopy plus mechanical lithotripsy through T-tube sinus. PMID- 12536713 TI - [Value of transvaginal sonography in diagnosis of placenta previa]. AB - OBJECTIVE: To investigate the value of transvaginal sonography (TVS) in diagnosis of placenta previa. METHODS: Transabdominal sonography(TAS) and TVS localization of the placenta were performed in 78 pregnant women after 35 week's gestation. It was suspected of having placenta previa when the placental edge appeared to be over or within 2 cm in diameter of the internal cervical os demonstrated by TAS or TVS. Final diagnosis depended on diagnosis at delivery. The diagnostic sensitivity, specificity, false-positive rate and coincidence rate were made in comparison between TVS and TAS. RESULTS: The sensitivity, specificity, false positive rate and coincidence rate of TVS were 100%, 90%, 10% and 93.6%, while that of TAS were 71.4%, 60%, 40% and 64.1%, respectively. The coincidence rate of TVS was better than TAS (p < 0.01). CONCLUSION: TVS is superior to TAS in diagnosis of placenta previa. PMID- 12536714 TI - [Gly40Ser mutation of glucagon receptor gene and NIDDM in Han nationality]. AB - OBJECTIVE: To ascertain whether Gly40Ser mutation in exon 2 of glucagon receptor (GCG-R) gene is associated with late-onset non-insulin-dependent diabetes millitus (NIDDM) in Chinese. METHODS: We selected 82 NIDDM patients and 136 controls of Han nationality in Hunan province, and detected Gly40Ser mutation using polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP). RESULTS: None of Gly40Ser mutation was found in all subjects. CONCLUSIONS: The mutation doesn't associate with NIDDM in Chinese, even though the mutation was reported to be associated with NIDDM in certain French and Caucasians, genetic heterogeneity of NIDDM among ethnic groups should be considered. PMID- 12536715 TI - [Two cases of child's dissecting aneurysm]. PMID- 12536716 TI - [The molecular mechanism of high glucose-induced insulin resistance in 3T3-L1 adipocytes]. AB - We investigated the cellular effect of high glucose using 3T3-L1 adipocytes on glucose transport activity, the expression of insulin signaling proteins and IRS1 tyrosine phosphorylation. Results showed that adipocytes treated with different high glucose (10, 15 and 25 mmol.L-1) for 24 hours showed to impair the basal and insulin-induced increase in glucose uptake in a dose-dependent manner and decreased significantly IRS1 tyrosine phosphorylation. High concentration of glucose produced opposite effects on IRS1 and IRS2: down-regulated IRS1 protein expression level and tightly up-regulated IRS2 contents. p85 and PKB were unaffected. Chronic exposure to high glucose can inhibit glucose uptake and induce insulin resistance. The mechanism may be involved in affecting the expression of insulin signaling peptides and tyrosine phosphorylation. PMID- 12536717 TI - [Expression of matrix metalloproteinases in metastatic lesions of cutaneous squamous cell carcinoma]. AB - OBJECTIVE: To investigate the expression of matrix metalloproteinases (MMPs) and its correlation with basigin/CD147 expression in metastatic lesions of cutaneous Sequamous cell carcinoma (SCC). METHODS: Expressions of MMPs and basigin/CD147 were examined by DAB and ALP immunohistochemical staining in 14 metastatic SSCs, and immunoblot analysis was used to observe the basigin/CD147 expression in metastatic samples. RESULTS: Normal skin and lymph nodes were basically negative for MMPs, but expressions of MMP-1, MMP-2, MMP-3 and MTl-MMP were increased in both tumor cells and stromal cells in metastatic SCC. Strongly positive or positive staining was observed in 4 or 8 of metastatic SCCs for basigin/CD147 in tumor cells respectively. Significant correlations were found between basigin/CD147 expression in tumor cells and MMP-1, MMP-2, MMP-3, MTl-MMP expression in surrounding stromal cells (p-values: 0.012, 0.024, 0.047 and 0.026, respectively). Immunoblot analysis revealed that basigin/CD147 expression increased in metastatic SCC. CONCLUSION: These results suggest that MMPs produced by both tumor cells and stromal cells play a role in metastasis of cutaneous SCC, and increased MMPs in stromal cells might be induced by basigin/CD147 expressed in tumor cells. PMID- 12536718 TI - [Inhibitory effect of yiqijiedu powder to telomerase and telomerase RNA in nasopharyngeal tumorigenesis of rats]. AB - OBJECTIVE: Telomerase activation is a common event in malignant transformation, and is associated with tumorigenesis. This has been clarified in nasopharyngeal carcinoma. YIQIJIEDU powder (Chinese traditional medicine) is used to treated the patients with nasopharyngeal carcinoma (NPC) after radio-therapy and patients with precancerous lesion of NPC. We investigated whether YIQIJIEDU powder inhibits the telomerase activity in nasopharyngeal carcinogenesis. METHODS: Nasopharyngeal carcinomas in rats were induced by N, N' Dinitrosopiperazine (DNP), and some of them were treated with YIQIJIEDU powder. Telomerase and telomerase RNA in the rats' nasopharyngeal tissue were assayed by using PCR ELISSA and Nested-PCR, and were pathologically diagnosed. RESULTS: YIQIJIEDU powder could inhibit telomerase activity and nasopharyngeal carcinogenesis in the nasopharyngeal tissue of the rats treated, but no telomerase RNA. CONCLUSION: The findings suggest that the anticancer effect of YIQIJIEDU powder links with their inhibitory effects on telomerase. PMID- 12536719 TI - [Listeria monocytogenes induces thymocyte apoptosis in mice]. AB - The murine thymocyte apoptosis induced by Listeria monocytogenes(LM) was detected with morphology, FCM, and DNA electrophoresis. The results were that LM elicited typical morphological changes of thymocyte apoptosis; the typical apoptosis peak was displayed with FCM, and typical "ladder pattern" with agarose gel electrophoresis. The apoptotic cells were found at 8 h after the mice had infected LM and reached climax at 48 h. The thymus weight significantly reduced at 16 h, and reached the lowest at 48 h after the mice had infected LM. The percentage of apoptotic cells was raised with the increasing of LM. These results suggest that LM induces thymocyte apoptosis in dose- and time-dependent manner. PMID- 12536720 TI - [Alterations of glial fibrillary acidic protein in rat brain after gamma knife irradiation]. AB - OBJECTIVE: To study glial fibrillary acidic protein (GFAP) immunoreactivity in different time and water content of the rat brain treated with gamma knife radiotherapy and to understand the alteration course of the brain lesion after a single high dose radiosurgical treatment. METHODS: In the brains of the normal rats were irradiated by gamma knife with 160 Gy-high dose. The irradiated rats were then killed on the 1st day, 7th day, 14th day, and 28th day after radiotherapy, respectively. The positive cells of GFAP in brain tissue were detected by immunostaining; the water content of the brain tissue was measured by microgravimetry. The histological study of the irradiated brain tissue was performed with H.E. and examined under light microscope. RESULTS: The numbers of GFAP-positive astrocytes began to increase on the 1st day after gamma knife irradiation. It was enlarged markedly in the number and size of GFAP-stained astrocytes over the irradiated areas. Up to the 28th day, circumscribed necrosis foci (4 mm in diameter) was seen in the central area of the target. In the brain tissue around the necrosis, GFAP-positive astrocytes significantly increased (P < 0.01, compared with the control group). The swelling of cells in irradiated region was observed on the 1st day; after irradiation endothelial cells degenerated and red blood cells escaped from blood vessel on the 7th day; leakage of Evans blue dye was observed in the target region on the 14th day. There was a significant decrease of specific gravity in the irradiated brain tissue the 14th and 28th day after irradiation. CONCLUSION: The results suggest that GFAP can be used as a marker for the radiation-induced brain injury. The brain edema and disruption of brain-blood barrier can be occurred during the acute stage after irradiation. PMID- 12536721 TI - [Expression and cloning of two novel genes induced by ischemia and reperfusion]. AB - OBJECTIVE: To identify and clone the differentially expressed genes in brief ischemia and reperfusion myocardium. METHODS: Ischemia and reperfusion were induced by repeated brief ligation of the porcine left anterior descending coronary artery. Total RNA which was isolated from myocardium subjected ischemia and reperfusion was used for mRNA differential display. After cloning and sequencing the cDNA fragments which showed change in expression, their expression were further confirmed by Northern-Blot analysis. RESULTS: Two differentially expressed cDNAs (W12 and W28) were identified and cloned. Their expression were subsequently confirmed to be truly differentially expressed. The expression of both genes in ischemia and reperfusion myocardium was obvious higher than that in nonischemia and reperfusion: W12 expression level was 2-fold (P < 0.05), and W28 expression level 1.9-fold (P < 0.05). In addition, mRNAs of W12 and W28 were existed in all tested organs including heart, liver, lung, kidney, spleen, intestine, brain and skeletal muscle. DNA sequencing analysis showed that there was no homology between W12, W28 and known genes, implying that they would represent novel gene respectively. CONCLUSION: Two novel genes induced by ischemia and reperfusion are identified, cloned and confirmed. PMID- 12536722 TI - [Protective effect of ginsenosides on human peritoneal mesothelial cell against lactate-based peritoneal dialysis solutions--induced injury in vitro]. AB - OBJECTIVE: The aim of this research was to observe the influence of ginsenosides (GS) on viability and proliferation of human peritoneal mesothelial cell (HPMC) which were inhibited by L-PDS in vitro. METHOD: Mesothelial cells were isolated from human omental specimens by trypsin disaggregation and established a stable HPMC culture model. The viability and capacity of proliferation of HPMC were assessed by lactate dehydrognase (LDH) release and tetrazolium salt colorimetry assay (MTT assay). RESULTS: Compared with the control group, the release of LDH in L-PDS groups (2.5%, 4.25% glucose) significantly increased. But there was no difference between the two glucose concentrations. As HPMC pre-exposed to L-PDS prolonged, the viability of cells decreased gradually. GS decreased the release of LDH and increased the cells viability (P < 0.01), as well as enhanced the proliferation of HPMC. CONCLUSION: The results suggest that GS has some protective effects on cell viability and proliferation of HPMC inhibited by L-PDS in vitro. PMID- 12536723 TI - [Culture and characterization of human peritoneal mesothelial cells]. AB - OBJECTIVE: To establish a reproducible model for culture of human peritoneal mesothelial cells (HPMCs), and observe the expressions of extracellular matrix (ECM) protein and interleukin-8 (IL-8) in HPMC. METHODS: Mesothelial cells were isolated from human omenta by trypsin EDTA disaggregation. HPMCs were identified by morphology and streptomyces protein-peroxidase (SP) method. In the same way, expressions of fibronectin, collagen type III, V and transforming growth factor beta 1 (TGF beta 1) were measured. Expression of IL-8 mRNA and FN mRNA in the HPMCs was detected by reverse transcription (RT) and polymerase chain reaction (PCR) amplification. RESULTS: Confluent HPMCs appeared multipolar under microscope; numerous surface microvilli and an abundant endoplasmic reticulum were observed. Cultured HPMCs coexpressed cytokeratin and vimentin, and synthesized fibronectin, TGF beta 1 and collagen types III (instead of type V). Expressions of IL-8 mRNA and FN mRNA were also observed in HPMCs. CONCLUSION: Establishment of cultured HPMCs model will provide the basis of the research in preventment of peritoneal fibrosis and roles of IL-8 in peritoneal dialysis. PMID- 12536725 TI - [Expression of platelet-derived growth factor-b and transforming growth factor beta 1 in the alveolar macrophages of allergic rats]. AB - OBJECTIVE: To investigate the role of alveolar macrophages in airway remodeling in asthma. METHOD: Sixteen Wistar rats were divided into two groups: control group and asthmatic group. The expressions of platelet-derived growth factor-b (PDGF-b) and transforming growth factor-beta. (TGF-beta 1) in the alveolar macrophages of allergic rats were detected by immunohistochemistry and in situ hybridization. RESULTS: The expression of TGF-beta 1 in alveolar macrophages in allergic group was increased in both protein level (P < 0.05) and TGF-beta 1 mRNA(P < 0.01); but the expression of PDGF-b had no difference between the two groups. CONCLUSION: The alveolar macrophages may involve in the airway remodeling of asthma. PMID- 12536724 TI - [Expressions of p-MAPK, cyclin D1, p53 protein and their relationship in osteosarcoma]. AB - OBJECTIVE: To detect the expressions of p-MAPK, cyclin D1 and p53 protein, and investigate their relationship in osteosarcomas. METHODS: SP immunohistochemical technique was used to detect the expressions of of p-MAPK, cyclin D1 and p53 protein among the 58 osteosarcomas and 14 osteoid osteomas samples. RESULTS: The positive rates of p-MAPK, cyclin D1 and p53 protein were 77.6% (45/58), 60.3% (35/58) and 44.8% (26/58) respectively in 58 osteosarcomas, and 21.4% (3/14), 21.4% (3/14) and 7.1% (1/14) in 14 osteoid osteomas. Their positive rates and expressive intensities in osteosarcomas were higher than those in osteiod osteomas. Their positive correlation was significantly observed between p-MAPK and cyclin D1 proteins (P < 0.01), and wasn't observed between p-MAPK and p53 proteins (P < 0.05) in osteosarcomas. CONCLUSIONS: MAPK phosporylation may be one of causes for activation of cyclin D1, which play an important role in osteosarcogenesis. The mutation of p53 oncogene may not result in osteosarcogenesis by MAPK signaling pathway. Detecting p-MAPK by immunohistochemistry may be one of diagnostic indexes for osteosarcoma and benign osteoma. PMID- 12536726 TI - [Effect of NOS positive neurons in rats with learning-remembering barrier]. AB - To study the variation of nitric oxide synthase (NOS) neurons in rats with learning-remembering barrier by immunohistochemistry and histochemistry. The number of NOS neurons decreased apparently in hippocampus CA1-4 of rats with learning-remembering barrier, there were lots of new NOS neurons in dentate gyrus granular cell grays and pear-shaped zone's cortex. It is suggested that NOS neurons of hippocampus exert very important action in rat's learning remembering; new NOS neurons in pear-shaped zone cortex and dentate gyrus granular cell grays may be a compensative reaction of the learning-remembering barrier, and they might exert important function in following recovery of learning-remembering function. PMID- 12536727 TI - [Analysis of amino acids and vitamins in ginkgo biloba leaves]. AB - The contents of amino acids and 10 kinds of vitamins in ginkgo biloba leaves were analyzed via HPLC. The results shows that it contains 92.26 mg amino acids per gram leaves the rate of essential amino acids amount to total amino acids amount is 46.9% and the contents of Vc, Vpp, VE, VB6, PGA is 81.47 mg, 9.43 mg, 4.09 mg, 2.90 mg, 1.69 mg per gram leaves respectively. The present effective data will be helpful for further research in its chemical composition and comprehensive utilization. PMID- 12536728 TI - [Simultaneously analysis of 15 parameters in the patients with the liver-blood deficiency syndrome]. AB - OBJECTIVE: Using HPLC-ED or RIA, we determined simultaneously 15 indexes in 27 patients with the liver-blood deficiency syndrome (LDBD). By means of multivarivate hierarchical cluster analysis and selections of typical variate, the results showed that 15 indexes were classified into 5 groups, and the typical variates of each groups were NE, T3, TXB2, ALD and cGMP. It suggests that LBDS has some pathopysiological characteristics such as decreased functions of sympathetic nerve activation, lower T3 syndrome, imbalance of the active substance regulating cardiovascular function and metabolism of salt and water, and abnormalities second signal substance in cellular membrane. PMID- 12536729 TI - [Effect of low molecular weight heparin on unstable angina]. AB - OBJECTIVE: To evaluate the effect of low molecular weight heparin (LMWH) and verus urokinase (UK) on unstable angina (UA). METHODS: Sixty seven patients with UA were divided in two groups by a single blind, controlled, randomized method: LMWH (anti-thrombi) group and UK (thromblytic therapy) group. Total ischemia load in 24 hours ambulatory ECG, mortality and cases of acute myocardial infarction (AMI) in 30 days were employed to compare the effect and safety between two groups. RESULTS: There was no significant difference in decreasing attacks of angina pectoris and total ischemia load between two groups. The UK group had two cases of AMI and five cases of complications of bleeding (two cases of epistaxis, two cases of bleeding gums, one case of hematochezia). CONCLUSION: Antithrombin therapy is prior to thromblitic therapy, treatment of LMWH is safer than that of low dose UK in UA. PMID- 12536730 TI - [Clinical observation of combined therapeutic effect of prostaglandin E1 and mecobalamin on diabetic peripheral neuropathy]. AB - OBJECTIVE: To observe the clinical effect of combined treatment of prostaglandin E1 and mecobalamin on diabetic peripheral neuropathy(DPN). METHODS: Seventy two patients of DPN were divided into 3 groups, they were given the drug of prostaglandin E1 (PGE1), mecobalamin, PGE1 plus mecobalamin (combined therapeutic group) and compared the therapeutic effects respectively. RESULTS: The improvement of DPN symptoms and nerve conducting speeds of combined therapeutic group was obviously better than that of the single PGE1 group or mecobalamin group(P < 0.01). CONCLUSION: Combined therapy of PGE1 and mecobalamin is better than each of single drug for improvement of DPN symptoms. PMID- 12536731 TI - [Clinical classification and the operative curative effects of microendoscopic discectomy in the elderly lumbar disc herniation]. AB - OBJECTIVE: To study the value of the classification of the elderly lumbar disc herniation(LDH) as an indication of microendoscopic discectomy(MED). METHODS: Based on the pathological features of the elderly LDH, eighty-one patients over fifty years old (Group A) were proved to be intraspinal complications by traditional operation and were divided into I and II types which were compared with what were seen in the operation. Sixty-two patients over 50 years old (Group B) were treated with MED and reviewed retrospectively, and their curative effects were compared with their types. RESULTS: A general intraspinal narrowness rate of Group A was 62.07% in Type I, 107.69% in Type II (P < 0.01). Group B was all followed up (mean 10 months). According to the macnnab criterion, the satisfactory rate was 100% in Type I, 75% in Type II (P < 0.05). CONCLUSION: The clinical classification of the elderly LDH makes a significant effect on the treatment of MED, can be used as an indication of MED, and can be popularized in clinic. PMID- 12536732 TI - [Study of intraoperative mitomycin C in preventing recurrence after pterygium surgery]. AB - OBJECTIVE: To explore the feasibility of intraoperative application of mitomycin C (MMC) in pterygium and observe the ultrastructure change of pterygium with or without intraoperative application of MMC. METHODS: Sixty two eyes of 57 patients with primary pterygium were randomly divided into MMC group and control group, 29 patients (32 eyes) in MMC group underwent transposition with intraoperative application of MMC (0.4 mg.ml-1 for 1 minute), whereas 28 patients (30 eyes) in control group underwent transposition. The recurrence rate, complications and side effects were observed and compared between the two groups. The ultrastructure of pterygium with or without intraoperative application of MMC was studied with transmission electron microscopy. The mean follow-up was(7.6 +/- 3.4) months (1-13 minths). RESULT: Eleven eyes of the 30 eyes in control group (36.%) showed recurrence, whereas only 2 of 32 eyes (6.3%) in MMC group showed recurrence (P < 0.005). No severe complication and side effect appeared during the follow-up. Most of fibroblasts in pterygium that underwent intraoperative MMC occurred the change of rough endoplasmic reticulum (RER) dilatation. CONCLUSION: Intraoperative MMC appears to be an effective treatment in preventing recurrence of primary pterygium. The fibroblasts in pterygium have RER dilatation after application of MMC for 1 minute during the operation. PMID- 12536733 TI - [Effects of katamine on cardiopulmonary bypass-induced interleukin-6 and interleukin-8 response and its significance]. AB - OBJECTIVE: To investigate the effects of ketamine on cardiopulmonary bypass induced IL-6 and IL-8 response. METHODS: Twenty-four adult patients undergoing cardiac valve replacement were divided into control and ketamine group (n = 12). In the ketamine group, 1 mg.kg-1 ketamine was infused intravenously at the beginning of operation and CBP. Plasma levenof IL-6 and IL-8 were measured at different time after induction of anesthesia. RESULTS: In both groups, the levels of IL-6 and IL-8 increased significantly compared with those at the time before CBP (P < 0.05), but compared with those in control groups, IL-6 and IL-8 levels decreased obviously in ketamine group(P < 0.05). CONCLUSIONS: Ketamine is effective to reduce cardiopulmonary bypass-induces IL-6 and IL-8 response and protect reperfusion myocardium. PMID- 12536735 TI - [Cardioprotective effect of exogenous phosphocreatine in patients undergoing open heart surgery]. AB - Twenty patients with rheumatic heart valve disease undergoing valve replacement were randomly divided into 2 group (n = 10): phosphocratine (neoton) group (group CP) in which 4.0 g neoton was used intravenously 2 days before operation and control group (group C) in which 5% G. S instead of neoton. The results showed that spontaneous recovery rate of heart beat in group CP was significantly higher than that in group C; requirement of inotropic drugs, serum MDA value and ultrastructural changes of myocardium were less in group CP than those in group C. There were no differences in release of myocardial enzymes and myocardial ATP content between 2 groups. It is suggested that neoton is an effective cardioprotective drug. PMID- 12536736 TI - [CT diagnosis of maxillary mucoceles]. AB - CT signs of thirty-six cases of maxillary mucocele surgically and pathologically proved were analyzed. 20 of 36 cases revealed as low density lesion. 16 of 36 cases revealed as isodensity or high density lesion. They were morphologically classified as three types: nodular type, inflammatory type, bony defect type. Cystic morphology and fluid density within the lesion obviously indicated mucocele. Differential diagnosis should be made between mucocele and polyps, when the lesion revealed as soft tissue density. Inflammatory type need to be distinguished from maxillary inflammation. And bony defect type should be distinguished from the benign or malignant tumor of maxillary sinus. The islet sign and bone sclerosis are considered to be valuable for the differential diagnosis. PMID- 12536737 TI - [Family support and quality of life among hemodialysis patients]. AB - OBJECTIVE: Sixty chronic renal failure patients who had received hemodialysis for 3 months or longer were interviewed with the Quality of Life Questionnaire and Social Support from Family Scale. The results were that the hemodialysis patients had a lower of quality of life and there was a positive correlation between family support and the patients' quality of life. The results suggest that both the patients' condition and their family background should be estimated so as to improve the quality of life of the chronic renal failure patients who had been received hemodialysis. PMID- 12536738 TI - [Expression of bcl-2 protein in uterine leiomyomas and normal myometrium]. AB - OBJECTIVE: To study the expression of the apoptosis regulatory protein bcl-2 in the uterine leiomyomas and in the normal myometrium. METHODS: The expression of bcl-2 protein was examined by immunohistochemical ABC staining in 40 cases of uterine leiomyomas and normal myometrium. RESULTS: bcl-2 expression was higher in the uterine leiomyomas than that in the normal myometrium, both in the proliferative phase and in the secretory phase of the menstrual cycle (P < 0.01); bcl-2 expression in leiomyomas in the secretory phase was stronger than that in the proliferative phase of the menstrual cycle (P < 0.01). CONCLUSIONS: The increase of apoptosis inhabiting gene bcl-2 may play an important role in the growth of the uterine leiomyomas. This action may be regulated by progesterone. PMID- 12536739 TI - [Treatment of cervical disc herniation with percutaneous arthroscopic microdisectomy]. AB - OBJECTIVE: To study the indications and efficacy of percutaneous cervical arthroscopic microdiscectomy (PCAD). METHODS: Twenty eight patients with cervical discs herniation or disorders received endoscopic spine surgery, 37 intervertebral cervical discs underwent partial nuclectomy or total nucleotomy; the efficacy and complication were evaluated in three months after operation. RELUT: Good/excellent results were obtained in 23/28(82.2%) cases according to MacNab criteria; no serious complication was found; the cervical stability did not decrease in most cases. CONCLUSION: PCAD is a safe, accuracy and rational method in diagnosing and treating herniated cervical disc and discogenic disorders. PMID- 12536740 TI - [Diagnostic and therapeutic value of fiberbronchoscope for the adult in the children with respiratory diseases]. AB - OBJECTIVE: To understand the diagnostic and therapeutic value of fiberbronchoscope for the adult in the children with respiratory diseases. METHODS: The results of diagnostic and therapeutic use in 107 child cases using fiberbronchoscope for adult were analysed. RESULTS: Pathogenic diagnosis was acquired in 107 cases; 17(17/21) cases of bronchial foreign bodies were extracted; 22 cases underwent bronchoalveolar lavage locally. CONCLUSION: Various type of BF of fiberbronchoscope for adult can be used in children at various ages, and to diagnose the child's diseases. Fiberbronchoscope for adult is safe and effective in the removing of foreign body and reliable in the treatment of atelectasis caused by inflammation, post-operation, and aspiration of foreign bodies. Fiberbronchoscope for adult can replace fiberbronchoscope for child unless the latter is used generally. PMID- 12536741 TI - [Facial nerve dysfunction after various surgical managements for benign parotid tumor]. AB - OBJECTIVE: To observe the incidence of facial nerve dysfunction following parotidectomy and the relationship of the extent of parotid gland resection and the histopathology types. METHODS: Clinical observation of 99 patients who underwent parotid surgery from 1996 to 2000 was studied to analyse the relation between facial nerve dysfunction and the extent of parotidectomy with House Backmann grading system. RESULTS: The overall incidence of facial dysfunction (HB > 1) was 36.4% for temporary and 3% for permanent dysfunction; Most of the dysfunction were partial and most concerning the marginal mandibular branch (34/99). The temporary facial dysfunction rate in total parotidectomy is higher than that of superficial and local parotidectomy. CONCLUSION: The dysfunction of facial nerve branches is correlated with the surgical managements. The most facial never dysfunction is temporary. The size of the lesion and the histopathology types will influence the choice of surgical managements. A proper surgical managements would reduce the incidence of facial dysfunction. PMID- 12536742 TI - [Clinical use of ultrasonic in diagnosis of ascites]. PMID- 12536743 TI - [Effect of dextran combination with flurosemide on nephrotic syndrome]. AB - OBJECTIVE: To compare the diuretic effect of coadministration of dextran and flurosemide with that of coadministration of Albumin and flurosemide on primary nephrotic syndrome. METHODS: In a double-blind placebo-controlled study, eighteen primary nephrotic syndrome patients on standard sodium chloride intake, random by divided into three groups and received by intravenous administration for 60 minutes for three days (a) FU (1 mg.kg-1) combination with a sham infusion, (b) FU (1 mg.kg-1) combination with 50 ml of 20% solution of Albumin, or (c) FU (1 mg/kg) combination with 250 ml dextran 40. Urinary volume, sodium and plasma atrial nartiuretic peptide concentration were assessed. RESULTS: The results showed that administration of FU alone increased mean cumulative urinary sodium and volume excretion as compared with the administration of sham infusion without treatment (P < 0.05). The administration of FU and albumin or FU and dextran 40 caused an even more marked increase of urinary sodium and volume excretion (P < 0.01), as compared with the administration of sham infusion without treatment. Plasma ANP increased significantly on both albumin infusion days and dextran 40 infusion days (P < 0.05). CONCLUSION: Coadministration of albumin and FU or dextran 40 and FU can increase the urinary volume and urinary Na and ANP significantly, dextran 40 can take the place of albumin because there was no difference between the administration of FU and albumin and FU and dextran in UV, Una and ANP. PMID- 12536744 TI - [Effect of artificial liver support system on the patients with severe hepatitis]. AB - OBJECTIVE: To study the effective treatment for severe hepatitis. METHODS: The severe hepatitis patients were treated by artificial liver support system on basis of the generalized treatment. RESULTS: After treatment the level of serum total bilirubin was significant decreased (P < 0.01); the level of aminotransferase were significant decreased (P < 0.01); the prothrombin time was significantly shortened(P < 0.05). The survival rate of severe hepatitis was significantly increased(72%, 31/43)(P < 0.05). CONCLUSION: The artificial liver support system can elevate the survival rate of severe hepatitis. PMID- 12536745 TI - [Perioperative observation of plasma endothelin in patients with congenital heart disease with pulmonary hypertension underwent open heart surgery]. AB - To elucidate the correlation between endothelin(ET) and pulmonary hypertension, plasma ET was measured perioperatively by radioimmunoassay in 22 patients with pulmonary hypertension who underwent open heart surgery(PH group) in comparison with 16 patients without pulmonary hypertension(non-PH group) also underwent cardiopulmonary bypass(CPB) surgery. RESULTS: Plasma ET was significantly increased during CPB in both group, Plasma ET decreased rapisly and returned to preoperative level within 6 hours after operation in the non-PH group. In the PH group, however, plasma ET decreased slowly and did not return to preoperative level within 48 hours postoperation. PMID- 12536746 TI - [Diagnostic value of measuring resistance of airway in the bronchus diastole test for asthma]. AB - OBJECTIVE: To discuss the variation of resistance of airway (Raw) and the diagnostic value of the bronchus diastole test for asthma patient. METHODS: The forced expiratory volume of the first second (FEV1) and Raw were measured. RESULTS: 1. The positive rate of FEV1 increase after the test was 68% (20/31). 2. There was a significant difference in percentage of FEV1 increase after the test between positive group and negative group. 3. After the bronchus diastole test, the Raw decreased significantly in both groups. CONCLUSION: The decrease of Raw after bronchus diastole test is more valuable for the asthma diagnosis than that of FEV1. PMID- 12536747 TI - [Study of acute supratentorial and infratentorial localized lesions by transcranial Doppler]. AB - OBJECTIVE: To evaluate the correlation between increased intracranial pressure in supratentorial, infratentorial and TCD parameters of basal arteries. METHODS: Sixteen New Zealand white rabbits were randomly divided into two groups: supratentorial group and infratentorial group. Intracranial pressures were raised after inflation of a preplaced epidural balloons. TCD parameters of the BA, rMCA were examined at intervals of 15 minutes after inflation of the balloon. ICP, ASP, HR, R were also recorded simultaneously. RESULTS: 1. The normal spectra of BA were characterized by a high-resistance, low-flow waveform. 2. At the 0.4 ml, 0.5 ml balloon volume level, ICP in supratentorial group was significantly increased than that in infratentorial group; 3. The correlation between ICP and MCA's TCD parameters were better than that between ICP and BA's TCD parameters in the supratentorial group. In the infratentorial group, the correlation between ICP and BA's TCD parameters were better than that between ICPs and MCA's TCD. CONCLUSIONS: 1. There are intracranial pressure gradients in mass lesions, so fibroptic probe should be placed at the place where the mass lesion takes place. 2. To perform non-invasive ICP monitoring by TCD, the basal artery should be chosen according to the mass lesions. PMID- 12536748 TI - [A case report of diploetic malignant meningiomas]. PMID- 12536749 TI - [Clinical X-ray analysis for 20 atypical lung cancer]. AB - OBJECTIVE: To analyze clinical X-ray features of atypical lung cancer, expected to improve the diagnosis for atypical lung cancer. METHODS: To collect 20 cases with lung cancer from Jan. 1987 to Jan. 2000, those patients were diagnosed as pneumonia infiltration tuberculosis or lymphoma by X-ray and CT scanning firstly. To analyse retrospectly theses cases and contrast with operation and pathology. RESULTS: All patients were verified as lung cancer after following reexamine and by pathology. CONCLUSION: The cases with sigle pathy shadow in the lung fields could not exclude lung cancer when they had hemoptysis or had no typical pneumoma signs. PMID- 12536750 TI - [Diagnosis of subtypes and lateralization for primary hyperaldosteronism]. AB - OBJECTIVE: To evaluate the value of CT and posture stimulation test (PST) in the differential diagnosis and lateralization of primary hyperaldosteronism(PA). METHOD: Retrospective study was done based on the clinical data and pathological examinations of 22 patients with PA. RESULTS: The baseline plasma aldosterone concentration (PAC) of most aldosterone-producing adenomas (APAs) was over 250 pg.ml-1. In the diagnosis of APA, the sensitivity of CT was 75% and the specificity of CT was 100%; the sensitivity and specificity of PST were 87% and 100%, respectively. The examination results of CT and PST had no statistical significant correlativity (P > 0.05) and had bad consistency (P > 0.05), and the diagnostic efficiency of them had no significant difference (P > 0.05). CONCLUSIONS: The baseline PAC and the result of PST suggest as the best diagnostic index and methods of APA. Both CT and PST have high value in the differential diagnosis. Both CT and PST cannot be substituted for each other. PMID- 12536751 TI - [Reamed interlocking intramedullary nail in the treatment of tibial and fibular fractures]. AB - To analyse retrospectively the results of thirty-eight fractures of the tibial and fibular treated with reamedintramedullary interlocking nail from May 1997 to June 2000. The patients were followed for an average of 14.6(range: 6-32) months. The mean age was 32.8(range: 18-64) years, the sex distribution was 29 males and 9 females. 26 fractures were closed (One case was failure of plate and screws) and 12 were opened: 6 were graded I, 4 were II, 2 IIIa, according to Gustilo's classification. A solid union was achieved in all patients within a period of 6 months. There were no deep infections except one case of superficial infection. Cases of deformity or peroneal nerve injury were no noticed. It is believed that reamed intramedullary interlocking nail allows a stable and safety fixation in open or closed tibial and fibular fractures. PMID- 12536752 TI - [Effect of lithium chloride on insulin release and DNA synthesis of cultured human fetal islet cells]. PMID- 12536753 TI - [Clinical efficacy of the combination chemotherapy of high-dose epirubicin in non small cell lung cancer]. PMID- 12536754 TI - [CBF measured by SPECT and intelligence scale in attention deficit hyperactivity disorder]. PMID- 12536756 TI - Changes in the chemical industry: the perspective of a catalyst supplier. PMID- 12536755 TI - Determining the structure of active sites, transition states and intermediates in heterogeneously catalysed reactions. PMID- 12536757 TI - The specific contribution of phosphorus in dendrimer chemistry. AB - Besides properties commonly found for all types of dendrimers, phosphorus containing dendrimers possess some specific properties seldom or never found for other types of dendrimers. Emphasis will be put on these specificities. PMID- 12536758 TI - A potassium selective 'nanosponge' with well defined pores. AB - The novel spherical cluster-compound K2Na6[K20 [symbol: see text] Mo80 V22O282(SO4)12(H2O)66]. approximately 140H2O 1 has been obtained in a 'one-pot' reaction and exhibits several remarkable structural features: 30 open shell centers, i.e. 8 Mov and 22 VIV spanning an icosidodecahedron, the incorporation of 20 K+ cations capping 20 appropriate [M6O6] pores/rings present at the surface of the spherical cluster-anion, and an encapsulated aggregate of water molecules. PMID- 12536759 TI - [NC4H12]2[(UO2)6(H2O)2(SO4)7]: the first organically templated actinide sulfate with a three-dimensional framework structure. AB - [NC4H12][(UO2)6(H2O)2(SO4)7] is the first organically templated actinide sulfate with a three-dimensional framework structure; it has channels of dimensions 8.5 A x 8.5 A, running along the [001] direction, containing tetramethylammonium cations. PMID- 12536760 TI - An hermaphroditic [c2]daisy chain. AB - A cyclic dimeric daisy chain compound, which has been assembled from a disfunctional [2]rotaxane in a sequence of noncovalent and covalent synthetic steps, the most important of which is a bis-Wittig reaction, has been characterised by X-ray crystallography. PMID- 12536761 TI - Electrochemically controlled interactions between TTF-based dendrimers and an electron-rich oligomer. AB - Electrochemically controlled interactions have been shown to occur between TTF containing dendrimers 1 and 2 and the electron-rich oligomer 3. PMID- 12536762 TI - Polymorphism of pure p-tert-butylcalix[4]arene: conclusive identification of the phase obtained by desolvation. AB - In its pure form, the well-known host compound p-tert-butylcalix[4]arene forms at least two polymorphs; one of these phases is now identified as that formed upon guest removal. PMID- 12536763 TI - Self-assembled oligonucleotide-polyester dendrimers. AB - A new approach to the controlled synthesis of multicomponent dendrimers is presented, in which three oligonucleotide-dendron conjugates were synthesized using solid phase techniques and hybridized to create a second generation polyester dendrimer with DNA as a core and bearing two types of peripheral functional groups. PMID- 12536764 TI - Designing a non-volatile imaging switch for mass-persistent, chemically amplified photolithography: a model study. AB - An acid catalysed rearrangement that transforms a bicyclic lactone into a phenolic carboxylic acid has been tested for potential use in chemically amplified microlithographic imaging. PMID- 12536765 TI - A very bulky carboxylic perchlorotriphenylmethyl radical as a novel ligand for transition metal complexes. A new spin frustrated metal system. AB - Reaction of a carboxylic perchlorotriphenylmethyl radical with Cu2(O2CCH3)(4).2H2O using different molar ratios yields two different transition metal complexes with strong intramolecular antiferromagnetic interactions, which in the case of complex 2 follows a butterfly spin frustrated model. PMID- 12536766 TI - Mechanochemical assembly of hydrogen bonded organic-organometallic solid compounds. AB - Solvent-free reactions with molecular systems have been exploited to prepare hybrid organic-organometallic solids: grinding of the complex [Fe(eta 5 C5H4COOH)2] with solid bases B generates quantitatively the corresponding hydrogen bonded salts [Fe(eta 5-C5H4COOH)(eta 5-C5H4COO)][HB] (B = 1,4 diazabicyclo[2.2.2]octane, 1,4-phenylenediamine); gas-solid reactions are also possible with volatile bases. PMID- 12536767 TI - In control of the speed of rotation in molecular motors. Unexpected retardation of rotary motion. AB - Surprisingly, a new motor with a tetrahydronaphthalene upper part rotates slower than the original molecular motor with a tetrahydrophenanthrene upper part despite decreased steric hindrance. PMID- 12536768 TI - Chemoselective transfer hydrodechlorination of aryl chlorides catalyzed by Cp*Rh complexes. AB - An effective Cp*Rh catalyzed transfer hydrodechlorination of aryl chlorides was achieved with high tolerance towards a variety of functional groups using 2 butanol as a hydrogen source. PMID- 12536769 TI - The first observation of four-electron reduction in [60]fullerene-metal cluster self-assembled monolayers (SAMs). AB - Self-assembled monolayers (SAMs) of a mu 3-eta 2:eta 2:eta 2-C60 triosmium cluster complex Os3(CO)8(CN(CH2)3Si(OEt)3)(mu 3-eta 2:eta 2:eta 2-C60) (2) on ITO or Au surface exhibit ideal, well-defined electrochemical responses and remarkable electrochemical stability being reducible up to tetranionic species in their cyclic voltammograms. PMID- 12536770 TI - Thermally reversible C60-based donor-acceptor ensembles. AB - Diels-Alder cycloaddition of anthracene derivatives--bearing fused pi-extended TTFs--to C60 yielded thermally reversible donor-acceptor materials which function as fluorescence switches. PMID- 12536771 TI - Functionalized heteropolyanions: high-valent metal nitrido fragments incorporated into a Keggin polyoxometalate structure. AB - Three examples of nitrido-functionalized polyoxometalate species are reported, namely (n-Bu4N)4[PW11O39(OsN)] (1), (n-Bu4N)4[PW11O39(ReN)] (2), and (n Bu4N)3[PW11O39(ReN)] (3), which feature the incorporation of [OsVI identical to N]3+, [ReVI identical to N]3+ and [ReVII identical to N]4+ fragments, respectively, into the framework of a Keggin-type heteropolyanion. PMID- 12536772 TI - High efficiency dye-sensitized nanocrystalline solar cells based on ionic liquid polymer gel electrolyte. AB - An ionic liquid polymer gel containing 1-methyl-3-propylimidazolium iodide (MPII) and poly(vinylidenefluoride-co-hexafluoropropylene) (PVDF-HFP) has been employed as quasi-solid-state electrolyte in dye-sensitized nanocrystalline TiO2 solar cells with an overall conversion efficiency of 5.3% at AM 1.5 illumination. PMID- 12536773 TI - New routes to high nuclearity cages: a fluoride-based hexaicosametallic manganese cage. AB - The reaction of MnF3 with benzotriazole (BTAH) in hot MeOH gives the Mn(III) complex [Mn26O16(OH)10(OMe)6(F)10(BTA)20(BTAH)2(MeOH)13(H2O)] which displays hysteresis and temperature-independent magnetisation relaxation at low temperature. PMID- 12536774 TI - Synthesis of 2-silyl substituted phospharuthenocenes and an elaboration into the first phospharuthenocene-phosphine. AB - A [1,5] shift protocol transiently hindering the 2-position of phospholide anions provides an access to sterically unencumbered phospharuthenocenes and the first phospharuthenocene-phosphine. PMID- 12536775 TI - A cyclic hexacopper(II) fluoro complex that encapsulates two fluoride anions. AB - The complex [[Cu3(HpztBu)4(mu-pztBu)2(mu-F)2(mu 3-F)]2]F2 (HpztBu = 3[5]-tert butylpyrazole) has a cyclic, C2v-symmetric hexacopper core. The two non coordinated F- anions are encapsulated within cavities formed by three HpztBu ligands. PMID- 12536777 TI - Conformational landscape surfing induced by off-on pi-pi stacking in a porphyrin quinone dyad. AB - A covalently linked porphyrin-quinone dyad crystallizes with two orientations of the quinone, extended away from (off) and cofacial with the porphyrin macrocycle (on), which induce different conformations of the macrocycle and model the recently proposed structural effect of a nearby residue on the heme prosthetic group of a nitric oxide synthase. PMID- 12536776 TI - Syntheses and magnetic properties of hexanuclear [Cp2Mn3(L1)4]2 and octanuclear [Mn8(L2)12(mu 4-O)2] (L1 = 2-HNC5H5N, L2 = 2-NH-3-Br-5-MeC5H3N, Cp = C5H5). AB - The reactions of manganocene, Cp2Mn, with 2-aminopyridine (L1H) or 2-amino-3 bromo-5-methylpyridine (L2H) give the novel hexanuclear and octanuclear Mn(II) amido cage compounds [Cp2Mn3(L1)4]2 (1) and [Mn8(L2)12(mu 4-O)2] (2); magnetic measurements on which provide a rare insight into the magnetic properties of amido-bridged metal clusters. PMID- 12536778 TI - First total synthesis of the marine illudalane sesquiterpenoid alcyopterosin E. AB - The first synthesis of the marine illudalane sesquiterpenoid alcyopterosin E was accomplished through a concise ABC ring-formation strategy using a rhodium(I) catalysed intramolecular alkyne cyclotrimerisation as key connection. PMID- 12536779 TI - Synthesis and structure of a novel supramolecular Zn3Co8 assembly containing a linear Zn3 core. AB - Reaction of Zn(OAc)(2).2H2O (OAc = acetate) with LH2 in THF followed by reaction with Co2(CO)8 yields the novel supramolecular assembly Zn3L2(OAc)2[Co2(CO)6](4).0.5CH2Cl2, which was characterized by X-ray diffraction (LH2 = (CH2)3(N=CH-2-OH-5-C identical to CSiMe3C6H3)2 or N,N'-propylenebis(2 hydroxy-5-trimethylsilylethynl-1-aldimine). PMID- 12536780 TI - Charge-transfer band shifts in iron(III) hexacyanoferrate(II) by electro intercalated cations via ground state-energy/lattice-energy link. AB - It is currently important to achieve and understand adjustments of optical properties: "guest cation" induced CT spectral shifts in Prussian Blue are shown to be driven (via its specific effect on the Fe(CN)6 CT-donor entity) by the cation lattice-energy interaction, as inferred from microgravimetry of introduced alkali-metal ions, and from independent solubility correlations for other intercalated cations. PMID- 12536781 TI - A rare ether-bridged cobalt complex which gives rise to an unusual 'serpentine' metal-ligand binding motif. AB - An unusual metal-ligand binding motif is found in dimeric cobalt(II) complexes coordinated by diamidoether ligands that bridge the metals in a 'serpentine' fashion through the ether donors of the ligand backbone rather than the amido groups. PMID- 12536782 TI - Production, isolation and structural characterization of [92]fullerene isomers. AB - High-resolution 13C NMR has been used to structurally characterize a single isomer possessing C2 molecular symmetry as well as an inseparable mixture of other isomers of [92]fullerene, produced from dysprosium arc-burned soot, separated and isolated via multi-stage recycling HPLC. PMID- 12536783 TI - Novel chiral gallium Lewis acid catalysts with semi-crown ligands for aqueous asymmetric Mukaiyama aldol reactions. AB - Asymmetric Mukaiyama aldol reactions in aqueous media (water-ethanol = 9:1) were catalyzed by chiral gallium catalysts with semi-crown ligands to give aldol products with good yields, syn-diastereoselectivities and enantioselectivities. PMID- 12536784 TI - The first structural characterization and determination of the isomerization activation parameters of a chiral phosphatitanocene. AB - The activation parameters (delta G++298 = 11.5 (+/- 1.0) kcal mol-1, delta H++ = 16.3 (+/- 3.0) kcal mol-1, delta S++ = 16 (+/- 11) cal mol-1 K-1) have been determined for the rac to meso isomerization of a phosphametallocene, bis(3,4 dimethyl-2-phenylphospholyl)titanium dichloride, 2, which has been structurally characterized. PMID- 12536785 TI - Catalytic insertion of diazo compounds into N-H bonds: the copper alternative. AB - The complexes TpxCu (Tpx = homoscorpionate) catalyse the insertion of diazo compounds into nitrogen-hydrogen bonds of amines and amides, under very mild conditions, with quantitative yields being obtained with equimolar ratios of reactants. PMID- 12536786 TI - Novel bis-acac-O,O-Ir(III) catalyst for anti-Markovnikov, hydroarylation of olefins operates by arene CH activation. AB - A novel, thermally stable, homogeneous Ir catalyst for the anti-Markovnikov, hydroarylation of olefins is shown to operate by arene CH activation via the formation of a bisacac-O,O phenyl-Ir(III) species. PMID- 12536787 TI - A new method for the synthesis of hydrophobized, catalytically active Pt nanoparticles. AB - A single step method for the synthesis of catalytically active, hydrophobic Pt nanoparticles by the spontaneous reduction of aqueous PtCl(6)2- ions by hexadecylaniline molecules at a liquid-liquid interface is described. PMID- 12536788 TI - Photochemistry of single wall carbon nanotubes embedded in a mesoporous silica matrix. AB - By embedding single wall carbon nanotubes in a mesoporous silica matrix (SWNT@SiO2) the photochemical properties have been measured upon laser excitation at 266 nm; the SWNT@SiO2 exhibits long-lived emission (lambda em = 400 nm, tau = 0.95 microsecond), transient absorption (lambda max = 390 nm, tau = 11 microseconds) and is able to generate singlet oxygen in D2O. PMID- 12536789 TI - Novel coordination of dicyanamide, [N(CN)2]-: preferential binding of the amide nitrogen. AB - CoII[N(CN)2]2(H2BiIm)2, 1, and [CoII[N(CN)2](H2BiIm)2]Cl, 2 (H2BiIm = 2,2' biimidazole) have been structurally, spectroscopically, and magnetically characterized with both containing dicyanamides bound in unprecedented manners; namely, solely via the amide nitrogen for 1, and via an imide N forming 1-D helical chains for 2. PMID- 12536790 TI - Synthesis and structure of an asymmetric copper(I) dimer with two-coordinate and four-coordinate copper(I) sites. AB - An asymmetric copper(I) dimer Cu(1)2(pip)2 [pip = (2-picolyliminomethyl)pyrrole anion] with both two-coordinate and four-coordinate copper(I) sites was synthesized and studied by X-ray diffraction and DFT calculations. PMID- 12536791 TI - Supported ionic liquid catalysis investigated for hydrogenation reactions. AB - The concept of supported ionic liquid catalysis (silc) has successfully been applied to hydrogenation reactions, which significantly reduced the required amounts of ionic liquid phase and enabled the usage of fixed-bed technology; the resulting catalysts exhibited high activity and outstanding stability. PMID- 12536792 TI - Development of a new sterically protecting auxiliary of the metacyclophane type and application to unsymmetrical diphosphenes, 1,3-diphosphaallene and 1,4 diphosphabutatriene. AB - A new bulky bromobenzene of the metacyclophane type was converted to an unsymmetrically substituted 1,4-diphos-phabutatriene whose spin-spin coupling constant (3Jpp) turned out to be larger, for the first time, than the 2Jpp value of the corresponding 1,3-diphosphaallene. PMID- 12536794 TI - Vesicles with asymmetric membranes from amphiphilic ABC triblock copolymers. AB - A new amphiphilic ABC triblock copolymer (poly(ethylene oxide)-poly(dimethyl siloxane)-poly(methyl oxazoline)) has been synthesized and demonstrated to form vesicular structures with asymmetric membranes in aqueous media. PMID- 12536793 TI - Stable supramolecular dimer of self-complementary benzo-18-crown-6 with a pendant protonated amino arm. AB - The novel 4'-(ammoniummethylene)benzo-18-crown-6 cation was synthesized as a hexafluorophosphate salt and found to exist as a cyclophane-like dimer in the solid state, gas phase and in acetonitrile solution. PMID- 12536795 TI - Hybrids of silver nanoparticles with amphiphilic hyperbranched macromolecules exhibiting antimicrobial properties. AB - Hybrids of silver particles of 1 to 2 nm in size with highly branched amphiphilically modified polyethyleneimines adhere effectively to polar substrates providing environmentally friendly antimicrobial coatings. PMID- 12536796 TI - New chiral pi-conjugated polymers based on a (1R,2R)-diiminocyclohexane chiral unit with weak interchain pi stacking. AB - New chiral pi-conjugated polymers consisting of alternating conjugated segments and (1R,2R)-diiminocyclohexane units with C2 symmetry were prepared by a palladium-catalyzed coupling reaction, they exhibited very high specific optical rotations ([alpha] up to -3000 degrees) and strong Cotton effects ([theta] 10(6) deg cm2 mol-1). PMID- 12536797 TI - Formation of organometallic polymer nanorods using a nanoporous alumina template and the conversion to magnetic ceramic nanorods. AB - Polyferrocenylsilane nanorods were prepared using a porous anodic aluminium oxide template followed by chemical etching; pyrolysis was used to obtain magnetic iron oxide-containing ceramic nanorods. PMID- 12536798 TI - Direct organocatalytic aldol reactions in buffered aqueous media. AB - Organocatalytic cross-aldol reactions catalyzed by cyclic secondary amines in aqueous media provide a direct route to a variety of aldols including carbohydrate derivatives and may warrant consideration as a prebiotic route to sugars. PMID- 12536799 TI - Potential-controlled switching of 2-(2'-thienyl)pyridine adsorbed on Au(111). AB - Switching between three different adsorbed states was achieved using potential control to separately induce electrochemical behaviour associated with changes in binding and orientation of the thiophene and pyridine moieties of 2-(2' thienyl)pyridine adsorbed to Au(111) in neutral and basic solution. PMID- 12536800 TI - Switchable thiophene coordination in Ru(II) bipyridyl phosphinoterthiophene complexes. AB - Switching between P,S- and P,C coordination modes of 3'-phosphinoterthiophene to Ru(II) results in substantial differences in the electronic spectra and cyclic voltammetry of these complexes. PMID- 12536802 TI - Spin alignment of orthogonal pi-radicals of directly meso-linked porphyrin arrays. AB - High-spin states of poly pi-radical cations of directly meso-linked Zn(II)porphyrin arrays were observed by ESR spectroscopy because of the stabilization of the high-spin states by sterically orthogonal alignment of adjacent pi-radicals. PMID- 12536801 TI - Spin trapping of superoxide in the presence of beta-cyclodextrins. AB - The trapping of superoxide anion with DMPO and DEPMPO has been carried out in the presence of a methylated beta-cyclodextrin, Me-beta-CD; inclusion of the spin adducts in the cavity of Me-beta-CD resulted in a seven-fold increase of their half-life (t1/2 = 96 min for DEPMPO-superoxide spin adduct) and in their protection towards glutathione peroxidase (Gpx) and also ascrobate anion in the case of DEPMPO. PMID- 12536803 TI - A convenient palladium-catalyzed aerobic oxidation of alcohols at room temperature. AB - A simple Pd-catalyzed aerobic oxidation of benzylic and aliphatic alcohols to the corresponding aldehydes and ketones at room temperature is described. PMID- 12536804 TI - Catalytic asymmetric dihydroxylation of olefins using a recoverable and reusable OsO(4)2- in ionic liquid [bmim][PF6]. AB - The use of the solvent systems water/ionic liquid or water/ionic liquid/tert butanol provides a recoverable, reusable, robust and simple system for the asymmetric dihydroxylation of olefins, based on the immobilization of the osmium ligand catalyst in the ionic liquid phase. PMID- 12536805 TI - Osmium tetroxide-(QN)2PHAL in an ionic liquid: a highly efficient and recyclable catalyst system for asymmetric dihydroxylation of olefins. AB - In Os-catalysed asymmetric dihydroxylation using NMO as a co-oxidant, the combination of an ionic liquid and the new bis-cinchona alkaloid 2 generated in situ from (QN)2PHAL during reaction provided a simple and practical approach to the recycling of both catalytic components (osmium tetraoxide and chiral ligand). PMID- 12536806 TI - Interfacing supramolecular and macromolecular chemistry: metallo-supramolecular triple-helicates incorporated into polymer networks. AB - Metallo-supramolecular diiron(II) triple-helical cores based on pyridylimine ligands and decorated with acrylate or methacrylate units are copolymerised with methyl methacrylate (MMA) as major comonomer using AIBN. PMID- 12536807 TI - Versatile enantiocontrolled synthesis of (+)-fostriecin. AB - Fostriecin, a potent protein phosphatase inhibitor and antitumor agent, has been enantioselectively synthesized in naturally occurring form via a versatile route, which also allows one to secure all possible stereoisomeres of the C1-C13 fragment including the C11 stereocenter and the geometry of the delta 12-double bond. PMID- 12536808 TI - A novel dicationic phenoxaphosphino-modified xantphos-type ligand--a unique ligand specifically designed for a high activity, selectivity and recyclability. AB - A novel phenoxaphosphino-modified ligand has been prepared and successfully employed in the rhodium catalysed hydroformylation of 1-octene in ionic liquids showing unprecedented high selectivity and activity without detectable rhodium or phosphorus leaching during recycling experiments. PMID- 12536809 TI - Solid state 13C NMR studies of methane dehydroaromatization reaction on Mo/HZSM-5 and W/HZSM-5 catalysts. AB - Methane dehydroaromatization on Mo/HZSM-5 and W/HZSM-5 catalysts was studied by solid state 13C NMR spectroscopy, both variation of the state of transition metal component and products such as ethane, benzene, ethene adsorbed on or in zeolite were observed after high temperature (900-1000 K) reaction. PMID- 12536810 TI - Novel Zn/ZnI2-promoted cross-coupling of acrylic acid esters with arylaldehydes to alpha-aroyladipic acid esters. AB - Cross-coupling of alkyl acrylates and arylaldehydes was achieved by treatment with metallic zinc and zinc(II) iodide in DMF, giving dialkyl alpha-aroyladipates in moderate yields. PMID- 12536811 TI - Amino acid functionalisation of water soluble carbon nanotubes. AB - High solubility of SWNTs and MWNTs in water is obtained by organic functionalisation; derivatisation with N-protected glycine is also easily achieved. PMID- 12536812 TI - beta-Hydroxysulfoxides as chiral cyclic ketone equivalents: enantioselective synthesis of polysubstituted cyclohexanones, cyclohexenones and cyclohexenediones. AB - The beta-hydroxysulfoxide moiety, after oxidation to sulfone, acts as a masked carbonyl group in a cyclic system, opening an easy access to differently substituted enantiomerically pure cyclic ketones by means of aluminium-mediated conjugate additions, stereoselective reductions and elimination by retrocondensation in basic medium. PMID- 12536813 TI - Supramolecular graphyne: a C(sp)-H...N hydrogen-bonded unique network structure of 2,4,6-triethynyl-1,3,5-triazine. AB - The crystallization of 2,4,6-triethynyl-1,3,5-triazine (2) leads to a pi-stacked layered structure of a C(sp)-H...N hydrogen-bonded unique hexagonal network structure, which may be regarded as a supramolecular analogue of a hitherto unknown graphyne network: in-plane intermolecular interactions are short and linear H...N contacts (2.31 and 2.34 A) and the interlayer separation is 3.23 A. PMID- 12536814 TI - Ligand exchange reactions within the coordination sphere of a molybdenum eta 2(4e)-alkyne complex: the formation of an indole in a cascade reaction involving an alkyne and isonitrile ligands. AB - The exchange of phosphorus-containing ligands within the coordination sphere of molybdenum alkyne complexes is shown to be facile, a similar reaction with CNxyl (xyl = C6H3Me(2)-2,5) results in a remarkable coupling between the alkyne and three isonitrile ligands and C-H bond activation. PMID- 12536816 TI - Triarylation of eta 6-dihydronaphthalene-Cr(CO)3 complexes. AB - With ease, eta 6-dihydronaphthalene tricarbonylchromium(0) complexes undergo a triple arylation with concomitant ring closure, when submitted to Heck reactions under 'Jeffery conditions'. PMID- 12536815 TI - Unexpected formation and structural characterisation of a novel rhodium B12 analogue. AB - The metalation reactions of the 2,2'-bidipyrrin 4 with different rhodium(I) precursors yield the complexes 5 and 6 and the unusual corrinoid 7, depending only on the type of the ancillary ligand employed. PMID- 12536817 TI - Nucleophilic substitution of alkylchlorodihydro[60]fullerenes: thermodynamic stabilities of alkylated C60 cation intermediates. AB - Thermodynamic stabilities of alkylated C60 cations (RC60+) were determined based on the activation free energies of SN1 solvolysis of the title compounds, indicating that these cations have stabilities comparable to that of the tert butyl cation. PMID- 12536818 TI - Aqueous synthesis of III-V semiconductor GaP and InP exhibiting pronounced quantum confinement. AB - A mild aqueous synthesis route was successfully established to synthesize well crystallized and monodisperse GaP and InP nanocrystals, which were proved to exhibit pronounced quantum confinement by room-temperature UV/Vis adsorption and photoluminescence (PL) spectra. PMID- 12536819 TI - An artificial ionophore based on a polyhydroxylated steroid dimer. AB - The Na+ transporting properties of the first member of a new class of artificial ionophores, based on a C2-symmetric polyhydroxylated steroid dimer, are described. PMID- 12536820 TI - Evidence for an anomalous redox ionic pair between Ru and Mn in SrRu0.5Mn0.5O3: an X-ray absorption spectroscopy approach. AB - X-ray absorption spectroscopy studies of the polycrystalline SrRu0.5Mn0.5O3 have been performed at Ru, Mn-L2,3 edges. The 2p-->t2g related peak at the Ru-L2 edge is more intense than the Ru-L3 edge and a shift of the 2p-->eg related peak by approximately 0.8 eV to higher energy than that of the Ru(IV) compound, SrRuO3 is observed. In combination with a crystal field multiplet calculation approach, a possible anomalous change in the spectral features is explained based on the existence of a redox ionic pair involving Ru(IV)/Ru(v)<-->Mn(III)/Mn(IV). PMID- 12536821 TI - Absolute configuration assignment and enantiopurity determination of chiral alkaloids and coumarins derived from O- and C-prenyl epoxides. AB - A combination method of ozonolysis and chiral stationary phase (CSP)-GC-MS analysis has been developed to determine the enantiopurity values and absolute configurations of a range of alkaloid and coumarin hemiterpenoids derived from C- and O-prenyl epoxides. PMID- 12536822 TI - [Life with dignity and peaceful death]. PMID- 12536823 TI - [Actual situation and problems about catheter care in patients with benign intestinal failure under home parenteral nutrition (HPN)]. AB - It is important for patients with benign intestinal failure under HPN, such as those with inflammatory bowel syndrome and short bowel syndrome, to maintain his/her indwelling catheter patent for a long time. The results of catheter care affects their quality of life. We describe the results of 18 patients who were under HPN. Their average age was 35.2 (range 21-49) years. The underlying diseases were Crohn's disease (n = 13), short bowel syndrome (n = 3), Behcet disease (n = 1), and nutrition support for pre-operative state of lung transplantation (n = 1). A totally implantable subcutaneous infusion port (SIP) was used for 16 patients and Broviac's catheter for two patients. Fundamentally, the self-infusion method was selected and nocturnal cyclic TPN was performed. The period of HPN was a mean of 601 (range 73-1, 159) days. Seven catheters (4 patients) were exchanged due to occlusion or infection, and one due to infection of the subcutaneous pocket. The average life of the catheters was 415 +/- 320 days. Catheter related complications occurred 0.74/1,000 days of catheter in place. We conclude that HPN is managed safely and that it is useful to maintain the QOL of patients with benign intestinal failure. All patients were satisfied with HPN. PMID- 12536824 TI - [Experience of Aozora Medical Clinic in EMInet]. AB - We present here the EMInet (Electronic Medical Information network), a closed information network established by the City government and the Medical Association in Matsudo City, Chiba, Japan. In this network, medical information is shared by multiple certified medical facilities under the informed consent of the patients. The medical record is written only by the physician in charge of the patient and reserved in the server computer. The medical record cannot be rewritten or modified by anyone except the physician in charge of the patient. Other medical professionals can only have access to the record. In the emergency room of certified hospitals, a physician can open the patient's record by using the IC card carried by the patient. PMID- 12536825 TI - [Home care for terminal patients and information technology]. AB - An electronic medical record sharing system, which supports management of terminally ill patients at home, was developed in Matsudo City as a trial project. The system interconnects several facilities including four general practitioners (GPs), a visiting nurse center and a supporting municipal hospital. It provides the latest patients' information from GPs to the municipal hospital in case of the acute change of the patients' condition and their transfer. Using this system the discussion between GPs and visiting nurses became much more fruitful than we had expected, while some doctors did not use the system because of their computer literacy. Problems in the system security and privacy protection as well as clarification of users' responsibility remain to be solved. A more expanded system is now being developed as a new enterprise of Matsudo Medical Association following this system. PMID- 12536826 TI - [Home-medical-care and IT--for the further diffusion of the tele-health system]. AB - In this study, we examined the economic benefit of the tele-health system based on a field survey performed in Kamaishi City in Iwate Prefecture, Nishiaizu Town and Katsurao Village in Fukushima Prefecture, and Sangawa Town in Kagawa Prefecture. The economic benefit analyzed here was based upon willingness to pay (WTP). The cost items of the system included those of initial introduction and maintenance; the former included costs of equipment and software, whereas the latter the salaries of staff and others. We compared the benefit to cost ratio (B/C ratio) among these cases. We also discussed measures to further expand the tele-health system. PMID- 12536827 TI - [Reason why home hospice is not widespread in Japan]. AB - A home hospice care program was begun at Fukushima Rousai Hospital in 1987. Nowadays, fifty percent or more of patients who die of cancer use this program, and patients who die in their own homes have reached about 40 percent. Based on our experience, we analyzed the factors in why home hospice care is not widespread in Japan, and pointed out some factors as follows. The main factors are insufficient disclosure of information regarding diagnosis and disease progression, poor palliative care, and incompleteness of the visit medical treatment system. Other factors are doctors' zeal for cure, patients' and/or families' excessive expectation of recovery, insufficient numbers of doctors and nurses with knowledge and skills concerning the home hospice care, doctors' and/or families' conviction that death in the hospital is natural, lack of social systems that support the family, limitations in the use of medicine at insurance drugstores, and the fact that insurance cannot be adjusted for medical treatment in the patients' home. PMID- 12536828 TI - [Home medical care and long-term care insurance]. AB - The difference in medical care at home before and after the introduction of the Long-term Care Insurance is discussed and reported on the basis of my 20 years of experience in the home medical care. I appreciate the introduction of the Long term Care Insurance in terms of its effect in making the general public interested in nursing as well as in those who need care. On the other hand, since emphasis was placed only on the degree of demand for the care in evaluating the qualification of the users of nursing facilities under the Long-term Care Insurance, public interest was focused simply on the degree of demand for the care. In the meantime, the fundamental problem of how to institute medical care at home appears to be neglected. As a result, many of those whose demand for the care have been assessed low make claims and request re-assessment, which results in a lot of personnel and financial waste in office procedures. Moreover, the families of those with the care necessity grade of 1 or more tend to falsely feel that they have been given the right to use the facilities, which decreases their motivation toward the home care. In reality, most facilities are crowded with users and improvement of the facility does not catch up with the rapid increase of users. Needless to say, the present situation is attributed to the inadequate governmental policies taken at the time of introduction of the Long-term Care Insurance. Immediate measures for improving the situation are required. In the case of elderly insurance facilities, in particular, in addition to evaluation of the conditions for using the facility solely on the basis of the degree of demand for the care, their positioning as the rehabilitation facilities aiming at achievement of home care should be clarified like before the introduction of the Long-term Care Insurance. A certain limitation is necessary regarding the duration of the use of the facility. Other than that, various problems have been proposed. We hope to examine these problems as well and report the results. We believe the most important thing is to exercise our ingenuity in creating a system enabling medical care at home in each community. PMID- 12536829 TI - [Elderly diabetics and care insurance system--does the home care service contribute to the home medical treatment?]. AB - To investigate whether home medical care service based on the public care insurance system contributes to the home medical treatment of elderly diabetics, the following aspects were evaluated. 1. Did glycemic control improve? 2. Were the patients pleased with the results obtained? 3. Was a load to the home care of the family reducing? 4. Did dependence on medical treatment decrease? The investigation was carried out targeting 17 patients among elderly diabetic patients of 65 years of age for whom home care service was provided. 1. HbA1c level did not change after the start of the care service. 2. Some degree of patient's satisfaction was obtained. 3. The burden of the family concerning home care decreased. 4. Dependence on medical treatment (repeated visits or hospitalization) decreased. Therefore, it was suggested that the care service contributed to the home medical treatment of elderly diabetic patients. It was raised as a problem what correspondence to the cases those who dislike the home care service, or an authorization application. From now on, it is considered important to introduce a care service in which a dietitian, a nurse, a pharmacist, etc, are involved. PMID- 12536830 TI - [Discussion on care management operation of a visiting nurse--a case of increased ADL by the support of independent life]. AB - In addition to the visiting nursing service conventionally provided, the Department of Long-term Care Insurance Service of this hospital inaugurated the home care supporting service in April 2000. Senior citizens rated higher in the degree of necessity of care in the initial accreditation and in the renewal accreditation of the Long-term Care Insurance tend to have fewer changes in the services for the last two years. At present, care managers of various professions are involved in the home care supporting services and have no choice but to provide care in non-specialty areas. Under the situation, care management by the visiting nurse helped an elderly increase ADL and live on his own, and the case is introduced in this article. Mr. K.T. developed angina pectoris at the age of 76, had recurrences of complications and repeated transfers of hospitals and was eventually admitted to the hospital. Though he had declined muscular strength and ADL because of the long bed-ridden life, he was discharged from the hospital. Nursing services centered on visiting nursing were provided as the home care supporting service when home medical care for the patient was started. Since Mr. K.T. required medical management, he and his family members were not sure whether it is possible to provide care for him at home and required guidance about health/life and mental supports. Therefore, visiting nursing care was provided by a nurse to assess needs or condition of the person, which reduced anxiety and encouraged the person. As a result, ADL increased and the degree of necessity of care decreased from 4 to 2. This success is attributed to the visiting nurse's appropriate care management based on the medical expertise from the perspective of nursing and the introduction of necessary services at the necessary time based on the appropriate assessment of changes in the physical condition or willingness and the nursing condition of family members. Coordination with the staffs engaged in each service also contributed to the success. As shown by this case, it may be necessary for care managers who can exercise their specialty to be engaged in the service or to change care managers depending on the condition of the care service receiver or considering the specialty of care managers for the benefit of the care service receiver and for the improvement of the efficiency of the operations. PMID- 12536831 TI - [Home medical care using long-term care insurance--centered on home oxygen therapy]. AB - About two-thirds of the patients undergoing home oxygen therapy apply for the Long-term Care Insurance. Most of them are aged 75 or over but the degree of necessity of care is relatively low. Two-thirds of them apply for the insurance for the medical care-related reason, which are twice as many as those apply for the insurance because for the medical care-related reason. Medical cares covered by the Long-term Care Insurance include disease control by the visiting nurse, the assistance for the hospital visit using visiting care or day service/day care and respiratory rehabilitation using day care. Special elderly nursing homes are the good place to accept elderly people living alone. The merit of the Long-term Care Insurance is rapid information supply from various professionals to doctors. Since it is also possible to disseminate the doctor's instruction to each staff, it is possible to carry out disease control at home as if they were admitted to the hospital. For these reasons, the Long-term Care Insurance is extremely effective to prolong the period when patients undergoing oxygen therapy are treated at home or to improve their QOL. PMID- 12536832 TI - [Effectiveness of chemotherapy for outpatients with gastric or colorectal cancer]. AB - We retrospectively evaluated the efficacy of chemotherapy regarding symptom control, toxicity and discharge rate in 39 patients with gastric or colorectal cancer. Treatment consisted of TS-1 (n = 16), TS-1 + CPT-11 (n = 8), CDDP + CPT 11 (n = 5), paclitaxel (n = 8) and MTX + 5-FU (n = 4) for gastric cancer and 5-FU + l-leucovirin (n = 6), 5-FU + CPT-11 (n = 5), MMC + CPT-11 (n = 8) and 5-FU protracted continuous infusion (n = 5) for colorectal cancer. The rates of symptom improvement were the following: pain 60% (10/15), general fatigue 56% (5/9) and abdominal fullness 53% (8/15). 87% (34/39) of the patients were discharged from hospital and continued chemotherapy as outpatients grade 3 toxicities were the following: anemia 10.3%, nausea and/or vomiting 7.7%, diarrhea 5.1%. There was no treatment related death. The rates of outpatient based treatment duration improvement were the following: gastric cancer: 47.6%, colorectal cancer: 72%. These data suggest that these treatments for gastric and colorectal cancer are safe and improve the patients' QOL. PMID- 12536833 TI - [Experience with chemotherapy at the Pitie-Salpetriere Hospital, Paris, France]. AB - Thanks to a scholarship of the French government, in 2000 I had the possibility to work at the Medical Oncology Service (Professor D. Khayat) of the Pitie Salpetriere Hospital, Paris, France as an intern for 9 months. I describe the use of chemotherapy in France based on my experience. Chemotherapy for solid tumors is provided mainly on an outpatient basis. In the case of inpatients, the period is very short. The program works because of the patients' way of thinking, good supportive care, and a well-constructed visiting nursing system. Especially, administration of steroids, G-CSF and erythropoietin as a secondary prevention for myelosuppresion seemed very effective. This system would be helpful for chemotherapy provided on an outpatient basis in Japan. PMID- 12536834 TI - [Rules to carry out hepatic arterial infusion chemotherapy at home for patients with liver metastasis of colorectal cancer]. AB - We analyzed the effectiveness and adverse effects of hepatic arterial infusion chemotherapy (HAI) for patients with liver metastasis of colorectal cancer in order to clarify the rules of HAI. We provided HAI to 72 patients after hepatic resection and to 119 patients with unresectable liver metastases. The preventive effect on hepatic recurrence was recognized in the group administered more than 15 g of 5-FU (total dose). The response rate of patients with unresectable metastases was 60.3%. Adverse effects were observed in 65.8% of the patients. It is important to establish a follow-up system in each hospital to treat the patients as soon as possible when patients develop an adverse reaction. PMID- 12536835 TI - [Assistance for improving QOL of patients with terminal disease cared at home]. AB - A visiting nursing service was provided for a female patient with the terminal stage of transverse colon cancer. The patient, who strongly wanted to stay at home, was discharged from hospital under continuous subcutaneous injection of morphine hydrochloride in late April 2001. The visiting nurse supported her life including the procedures for the continuous subcutaneous injection for attenuating pain as the main symptom. As a result, her fear of pain was reduced and she became able to control pain by oral medication. She became able to walk to the rest room and take a shower and have increased ADL while regaining the strength of will. Family members were concerned with the potential sudden change in her conditions or intensification of pain at home but the worry was contained by understanding the procedures to follow in such cases. QOL can be improved even at the terminal stage if: 1. the patient understand the pathological condition; 2. the patient discloses his/her worry; 3. the patient can choose the way of living; 4. caregiver can cope with the change; 5. caregiver can maintain the nursing capability; 6. the medical provider's assistance system is established. PMID- 12536836 TI - [Medical network in home palliative care]. AB - Medical network in home palliative care for terminal cancer patients is more difficult compared to that in home care for patients with other diseases because of the following reasons: 1. only a little time for the rest, 2. difficulty of total pain control including physical pain; and 3. difficulty of family care. We think that in the case of home palliative care it is necessary to establish a closer connection and a relationship of mutual trust among medical coworkers. PMID- 12536837 TI - [A discussion of opinion survey among bereaved families of patients with terminal cancer under home care]. AB - Since its foundation, general medical care as well as home medical care for patients with terminal cancer are provided at the hospital. In order to clear the doubt if patients/family members feel secure or insecure about home medical care provided since the initial consultation till death at home only by the hospital staffs, the opinion survey was conducted for family members who were also member of the team. At the hospital, home medical care is initiated if the patient himself/herself accepts the pathological condition. It is impossible to continue such a medical care without establishing the reliable relationship between patients/family members. Medical providers like us are just the advisors and collaborators to help patients peacefully accept the terminal disease. This survey reminded us of the necessity of conducting the opinion survey so that medical care won't be centered on medical providers. PMID- 12536838 TI - [Standard indication of home oxygen therapy for terminal cancer patients]. AB - Home oxygen therapy for terminal cancer patients is not suitable for present standard. As the result of the questionnaire answered by home care doctors, many of them agreed to reconsider the present standard. We propose the following revision, 1. Standard that highly regard clinical symptoms, 2. subdivision of the fee, 3. addition of home care fee. PMID- 12536839 TI - [Roles of nursing counseling room in home medical care]. AB - The hospital inaugurated the service of visiting nursing in 1985 and the scope of the service was subsequently expanded to patients at the terminal stage and patients under artificial respiratory management. At present, however, the service of visiting nursing is restricted to the assistance to visiting medical care as a result of the diversification of the services of Nursing Counseling Room and the provision of better services at the visiting nursing station established in the city. The author reviewed the roles of the Nursing Counseling Room based on the experiences of discharge arrangements made in 2000 and 2001 in a patient with terminal disease, a patient with neurological refractory disease under artificial respiratory management and a child also under artificial respiratory management. Experiences in these patients made us realize the necessity of acting on the department of welfare of the municipal government, coordinating with the visiting nursing station, effectively holding conferences to discuss discharge and the necessity of the home doctor. We are determined to work to find solutions for these challenges. PMID- 12536840 TI - [Consideration concerning a care orientated medicine for the patient with terminal stage of cancer]. AB - Home care medicine plays an important role in the care of terminal patients with redundant diseases such as cancer. In this study, we propose a "simple-style medicine" consisting in the participation of both medical and co-medical staffs and the use of Kampo medicine. This system satisfied various and individual requests from terminal patients. We present 2 terminal patients for whom this "simple-style", home care-orientated medicine was effective. This system can contribute to the quality of life of patients who need palliative, home care medicine. PMID- 12536841 TI - [Assistance for improving QOL of patients using artificial respirator--a case of the very aged]. AB - The recent reform of the health insurance/Long-term Care Insurance resulted in the introduction of healthcare technology to home and patients can use even an artificial respirator in home medical care. However, both patients and family members have significant mental/physical burden, and in many cases, it may often be impossible to provide care to improve ADL/QOL. This time, a very old patient aged 88, who underwent artificial respiratory management, HOT, tracheotomy, tubal feeding, balloon catheterization and decubitus treatment among other medical cares, was discharged from the hospital and QOL was successfully improved by the care of family members and other various services. The success was mostly attributed to the endeavors of caregivers. In this article, we review the visiting nursing and assess what cares contributed to the improvement of QOL and discuss future assistance. PMID- 12536842 TI - [Progress of introduction of artificial respirator at home--the knowledge I have gained from a 19 year-old patient requiring a home respiratory management care and through the friendship with his family members]. AB - The patient developed right cerebellar hemorrhage at the age of 15 and spent four years at hospital. The patient and the family members had a strong wish to bring the patient back home and the patient was admitted to the hospital to undergo guidance about home medical care. The patient had paralysis of right upper and lower extremities and generalized strong ataxic motion and was under gastric feeding and had underwent tracheotomy to cope with hypoventilation, which required use of an artificial respirator at night hours. Since the patient was still as young as 19, we want to have the patient "go home" as they wanted. Fortunately, the mother who had to play key roles was still young and the parents worked at home. It didn't take long for them to master the techniques though they had various anxieties. It is considered important in the guidance on home medical care to what degrees the caregivers accept the condition of the patient and can imagine the long lasting care at home. We provided assistance while repeatedly interviewing with the patient and family members considering the significance to confirm their willingness and psychological condition. As a result, the patient was discharged from the hospital in two months. Now, two years have passed, and the patient is still cared at home and the possibility of the patient is sought for. The progress is reported here in this article. PMID- 12536843 TI - [Examination of living environment upon return to home for patients with cervical spinal cord injury--report of a case]. AB - Living environment is a problem for spinal cord injury patients upon their return to the society. There are also many instances of home care problems after hospital discharge. For cervical spinal cord injury patients in particular, appropriate intervention/assistance provided by the authorities in accordance with the level of residual function is essential, but there have been few reports on this issue. Here we report on a municipal facility for cervical spinal cord injury patients where a patient moved into upon hospital discharge that did not provide a suitable environment, and the burden posed on his family increased. The patient was a 55-year-old man with traumatic cervical spinal cord injury who was commuting to our rehabilitation center after hospital discharge. We investigated the environment in the municipal housing facility he had moved into, and examined the relationship between the level of cervical spinal cord injury function and ADL assessment before and after hospital discharge. The main problems were an increase in care provided by his wife due to unsuitable toilet facilities and insufficient public facilities concerning the living environment. In terms of ADL by FIM (functional independence measure), namely, toilet transfer, toilet motion and defecation control which he had performed independently at the time of hospital discharge, all had decreased to a level requiring assistance after he moved into the present apartment. Therefore, approach via welfare equipment compensation were undertaken. As a result, the burden for his wife diminished, but the ADL and FIM scores obtained were low. Thus, the living environment facilities for patients with cervical spinal cord injuries should be in accordance with the level of function and ADL, suggesting the necessity for the authorities to establish a cooperative system for running home facilities. PMID- 12536844 TI - [Changes in peripheral lymphocytes count in unconscious patients treated at home with Japanese herbal medicines (Hozai)]. AB - We investigated changes in peripheral white blood cells count, lymphocytes count, and serum albumin level in five unconscious patients administered Japanese herbal medicines (Hozai). The causes of unconsciousness were intracerebral hemorrhage in 3, cerebral infarction in 1, and craniopharyngioma in 1. The administered Hozai were Hotyu-ekki-to in 4 and Jyuzen-taiho-to in 1. To assess the effect of Hozai on the immune system of unconscious patients, we investigated these parameters before and after treatment. We observed normalization of WBC counts, increase of lymphocytes count, and unchanged levels of serum albumin after Hozai treatment. These findings suggest that Hozai may improve the immunological competence of unconscious patients. One of the problems affecting unconscious patients treated at home is infectious diseases, especially respiratory infections. It is considered that improvement of their immunological competence would contribute to prevent infections. These results suggest that Hozai treatment might be useful adjuvants to support the general condition of the patients treated at home. PMID- 12536845 TI - [The necessity of community rehabilitation for outpatients with a communication disorder]. AB - In order to explore the necessity of community rehabilitation, we conducted a questionnaire survey of patients with a communication disorder who had finished speech therapy at our hospital during the period of April 1998 to November 2001. A total of fifty-four subjects answered the questionnaire. The following observations were made based on the results. 1. Fifty-nine percent of the respondents was experiencing some degree of difficulties in their daily life, and facing some sorts of psychological and social problems. 2. Fifty nine percent considered community rehabilitation was necessary, while only one respondent was actually receiving it. 3. The majority of respondents wished there were speech therapists in their area to provide a variety of services such as recreation and rehabilitation. These results suggest the importance of speech therapy through local welfare and social care facilities. PMID- 12536846 TI - [Present situation of terminal care and home rehabilitation at Karuizawa Hospital]. AB - Rehabilitation is usually considered to have the connotation of regaining function or reintegration into society, and no direction or achievement regarding the time of involvement or details of request has been suggested for the terminal rehabilitation in the patients with cancer who can't expect to have any improvement. Dieatz defines four phases for the goal of rehabilitation in the patients with cancer; Preventive Phase, Recovering Phase, Supportive Phase and Alleviative Phase. This time, we encountered a couple of cases in patients with cancer: patients who gained QOL through developing environment and setting goals that enable them to maintain the active daily life (ADL) at the supportive phase; and patients who realized us when relieving the pain while losing self consciousness due to psychosomatic changes at the alleviative phase. In this report, we discuss what care can be provided at the phase when trust of the patient/family members is established, at the phase when care providers can grasp needs, and at the phase when the needs can be satisfied. We also report on how the patient and family member reacted to the direct hand massage provided by the physical therapist at the terminal stage. The rehabilitation system for home care at each phase is discussed in association with practical side and problems of the rehabilitation in the future terminal care. PMID- 12536847 TI - [Experiences in bereavement care for caregivers]. AB - PURPOSE: To investigate the mental and physical changes of bereaved caregivers. METHODS: We had 5 patients under residential care (1 with prostate cancer, 3 with urinary bladder cancer and 1 bedridden patient without malignant disease) and one month and one year after their death, we visited their family to provide bereavement care. RESULTS: Before the patient's death, all caregivers were uneasy and confused. At the deathbed, all of them were filled with deep grief and prostration. One month later, unrest and fatigue were reduced, all had the satisfaction of having taken care of their patients. One year later, only grief persisted and prostration was deserved in one caregiver. CONCLUSION: We provided bereavement care to 5 caregivers. It is suggested that in some cases long-term bereavement care is necessary. PMID- 12536848 TI - [Role of public hospitals regarding the support of patients at home]. AB - Eight sickbeds for the support of patients at home were established in Chiba Municipal Hospital in response to a request from the Chiba City Medical Association in 1997. These sickbeds were run by physicians and nurses in the hospital and nurses from Chiba City Public Health Center to promote a tight relation among the medical care, the health care and the public welfare systems. The role of the public health nurses is to receive and manage the documents of the patients from regional medical practitioners and to communicate with the related organizations. These sickbeds were set up to offer medical care, educational teaching to the family, and the social environment to support the patients at home. The patients, who were consulted by regional medical practitioners, were pre-registered by them. The length of stay at the hospital was limited to 2 weeks. The number of patients who were registered and hospitalized increased every year: thus 374 patients receiving treatment from 44 medical practitioners have already been registered. The average age of the patients was 81.1 and almost all of the patients were sick in bed for the whole day at home. Sixty-three patients were hospitalized making use of those sickbeds this year. Further promotion of the relation among the organizations involved in medical care, health care, and welfare is very important to support the patients to be able to stay at home. PMID- 12536849 TI - [Formulation of policies for promoting home medical care by a university hospital -report of planned and convened case review sessions]. AB - It is no exaggeration to say that the linkage among the persons in various specialties, medical staff including the home doctors and visiting nurses, and those related to social welfare is essential for supporting and promoting the terminal care at home of the patients heavily dependent on medical care, as well as the patients and their families with problems in the nursing ability. Good understanding of other specialties and compensation for each other are considered to enable effective service to the patients. At our hospital, we have planned and convened case review sessions for the past two years. The sessions were proved to have the following merits that helped creation of teamwork for smoothly promoting medical care at home: 1. Persons in each specialty became able to play respective roles properly; 2. Measures to be taken were revealed, thereby inducing changes in the subsequent care; 3. The efforts made in the community could be understood and the problems to be wrestled within the hospital were suggested; 4. Smooth cooperation with visiting nurses and home doctors was achieved; 5. Nursing from a broader point of view became possible; and 6. A place for identifying problems and tasks under the present situation was provided. PMID- 12536850 TI - [Introduction of a community network system to visiting care]. AB - A community-based electronic medical record sharing network system (EMI net) was developed in Matsudo City. This system facilitates communication between GPs and visiting nurses in charge of patients under homecare and it has proved useful from the view point of visiting nurses. For example nurses can read the information input by visiting GPs timely, share common view on the patients with GPs, and write a report illustrated with photographs at any time. Moreover, they can communicate with GPs through E-mail. On the other hand, EMI net remains to be improved in a few aspects, such as the need of a system to indicate a record has been read by GPs and to decrease the labor for entering data. We plan to increase the services provided by EMI net by enrolling other facilities. PMID- 12536851 TI - [Opinion survey among nursing students on family]. AB - Earlier discharges of patients from hospital are increasing needs for supports for home medical care, which is causing changes in the activities of medical providers for patients and family members. Meanwhile, it is suggested that the younger generation is more indifferent to family members along with the increase of nuclear families or households consisting of only one member. This time, the author conducted an opinion survey among nursing students for the purpose of understanding how nursing students think of the family and using the data as a reference for the education. The survey was conducted in FY2000 and FY2001 for the questions selected from the "National Lifestyle Popularity Survey". The results indicate that nursing students intend to have families while keeping working and are willing to fulfill their responsibilities as a family member of which relation is close. It is considered possible to guide nursing students to practice nursing respecting the position and feeling of the patient by letting them understand social changes and the individuality of the patient based on the way nursing students think of the family. PMID- 12536852 TI - [Improved ambulatory syringe pump system for pain control therapy at home]. AB - Terumo Corporation has been developing products for use at home, such as the Cafty pump, based on the opionions and suggestions received through Home-Joint. This time we improved the ambulatory syringe pump TE-361 PCA, so that patients could use it at home. RESULTS: We had patients 49 to 78 years of age use it, continuously, at an injecting speed of 0.1 to 1 ml/h for 5 to 26 days. Patients requiring large volumes of a medicine were offered two or more pumps pre-filled with the medicine. Drug transportation problems were not reported, because of the doctor in charge transported the medicine constantly. Terumo call center did not receive any incidence of trouble, and no trouble was reported in the collected questionnaire. From these facts, it was suggested that this apparatus can be used safely for in-home medical treatment. CONCLUSION: The burden of expenses when two or more pumps are required and installation of the back-up equipment, etc. are problems that should be coped with separately. Moreover, a cooperation system is required in preparation for troubles. We want to establish a system that can be used with confidence by the patients. PMID- 12536853 TI - [The present situation and subject]. AB - Terumo Corporation has been supporting the regional medical network, developing new products for home health care and providing service through the Home-Joint system in the fields of HPN, HEN, and HOT. In the chain of servicing, we are carrying out call center operations that can receive inquiries regarding machine operation and machine trouble from the patients, for 24 hours a day, all year round. Most inquiries are settled then and there, but some cases need further treatment. The principal cases are, Lowness mastery degree of patients or tenders, wants of knowledge of the person in charge of answering inquiries, and coming under the medical treatment. It is indispensable to connect closely with Hospitals, the visiting nurse, and agencies. Today's Subject: Pursuit of simplicity and safety of equipment based on On-Call experience. Thorough, careful confirmation for several days from the start. Standardization of On-call information and sharing with the agency and hospital. Advancement of knowledge for answering inquiries. Making a system to cope with medical facilities. PMID- 12536854 TI - Funding the essentials: the Australian Health Care Agreements, 2003-2008. AB - This editorial reviews a number of papers in a special issue of the Australian Health Review covering the Australian Health Care Agreements to be concluded by June 2003. These include a report on consultations by the Australian Healthcare Association with industry representatives from July-October 2002. For hospitals, the agreements will set the main financial parameters for the next five years. Apart from the quantum of Commonwealth grants, the issues seen as most important involved linkages with primary care providers and aged care facilities, the dominance of inpatient work in current arrangements, workforce planning and public/private sector relationships. The possibility of recent private health insurance changes reducing the sums available for public hospitals was noted. Some estimates are presented of the possible effects of private insurance reform, together with some data from a special AHA survey of public hospital activity this year. PMID- 12536855 TI - Renegotiation of the Australian Health Care Agreements 2003-2008: from a state's perspective. AB - The renegotiation process for the next Australian Health Care Agreements 2003 2008 presents an opportunity for State and Territory Governments to suggest reforms to improve acute and sub acute health services and health outcomes. Four key issues to Victoria are discussed, emergency departments and primary care interface, workforce planning, aged care and the continuum of care. PMID- 12536856 TI - The Australian Health Care Agreements--a teaching hospital perspective. AB - The Australian Health Care Agreements (AHCAs) are important for patients, health care professionals and hospital CEOs alike. The current agreements have not been fully effective in promoting necessary reforms, and this paper suggests priority areas for attention in the next round. Five areas for targeted reform are suggested. These are pharmaceuticals, workforce planning, continuum of care across settings, education and research, and safety and quality. It is time to give some radical thought to the role and scope of the AHCAs. Healthcare policy reform needs to be across the continuum of care. PMID- 12536857 TI - The next Australian Health Care Agreements: what clinicians want. AB - For the first time experienced clinicians are to be involved in the development of the Australian Health Care Agreements (AHCAs) (2003-2008). As a result doctors, nurses and allied health professionals are hoping that current impediments to necessary change will be removed. Numerous suggestions to improve the Agreement will come forward from the "front line". All clinicians will argue that the next Agreement must result in a marked increase in the funding available to public hospitals. The new Agreement must remove barriers hindering our capacity to integrate all of our healthcare services. Safe, appropriate and cost effective healthcare delivery must embrace a continuum of care involving patients and their primary care physician, community health services and hospitals. The Agreement must embody arrangements for markedly enhancing our public health efforts in the area of prevention of disease. Australia's clinicians are worried about inequity in terms of access and outcome for their patients and are willing to be partners in health care governance to improve the situation. PMID- 12536858 TI - The 2003-2008 Australian Health Care Agreements: an opportunity for reform. AB - The Australian Health Care Agreements govern the relationship between the Commonwealth and States about public hospital funding. The incentives enshrined in the Agreements can shape policy priorities. Although they provide for the largest specific purpose grant a State/Territory receives, the current negotiations should not be seen as providing the only opportunity for reform of health care for the next five years. This paper argues that the negotiations should focus on two key areas where Commonwealth-State frictions are high but reform is feasible. Specifically it is suggested that the Commonwealth should contribute its funding of public hospital inpatient services on a casemix basis, and secondly, should fund outpatient services directly. PMID- 12536859 TI - The 2003-2008 Australian Health Care Agreements--an industry perspective. AB - This paper presents a public hospital and health care industry perspective on the development of the 2003-2008 Australian Health Care Agreements. The Australian Healthcare Association conducted a national industry consultation exercise from June to September 2002 in the lead up to the development of the next round of agreements. While acknowledging that the size of the funding commitment from the Commonwealth to the states will be the central focus of negotiations, health industry representatives identified issues of equal importance. The AHCA's linkages with other health programs need to reflect that health care has moved beyond the confines of the hospitals. Adjustments and output targets need to provide incentives to improve and reform the industry. The success of private health insurance policy has not yet translated into benefits for the public hospital sector, and any funding contingencies between the two programs cannot be justified at this time. Special priority areas such as health workforce will need specific policy and program responses. PMID- 12536860 TI - Policy change and private health insurance: did the cheapest policy do the trick? AB - From the introduction of Australia's national health insurance scheme (Medicare) in 1984 until recently, the proportion of the population covered by private health insurance declined steadily. Following an Industry Commission inquiry into the private health insurance industry in 1997, a number of policy changes were effected in an attempt to reverse this trend. The main policy changes were of two types: "carrots and sticks" financial incentives that provided subsidies for purchasing, or tax penalties for not purchasing, private health insurance; and lifetime community rating, which aimed to revise the community rating regulations governing private health insurance in Australia. This paper argues that the membership uptake that has occurred recently is largely attributable to the introduction of lifetime community rating which goes some way towards addressing the adverse selection associated with the previous community rating regulations. This policy change had virtually no cost to government. However, it was introduced after subsidies for private health insurance were already in place. The chronological sequencing of these policies has resulted in substantial increases in government expenditure on private health insurance subsidies, with such increases not being a cause but rather an effect of increased demand for private health insurance. The paper also considers whether the decline in membership that has occurred since the implementation of lifetime community rating presages the re-emergence of an adverse selection problem in private health insurance. Much of the decline to date may be attributable to failure on the part of some members to honour premium payments when they first fell due. However, the changing age composition of the insured pool since September 2000, resulting in an increasing average age of those insured, suggests the possible reappearance of an adverse selection dynamic. Thus the 'trick' delivered by lifetime community ratings may not be maintained in the longer term. PMID- 12536861 TI - The price of choice: private health insurance in Australia. AB - Private Health Insurance (PHI) is an integral part of the financing of the Australian health care system. PHI is popular and has strong political support because it is perceived to give choice of access and responsiveness. However, in the past increasing premiums have led to a progressive decline in membership. A package of reforms by the Commonwealth Government in support of the private health insurance has reinvigorated the industry over the last three years. Some strategies for achieving a sustainable PHI industry are described. The key challenge is to control claims cost to maintain affordable premiums. Many techniques to do this compromise choice and challenge the very rationale for purchasing the product. Funds and providers will have to establish a new level of relationship to meet this challenge. PMID- 12536862 TI - Health insurance: managing risk in the private sector. AB - How does Australia, in both its private and public sectors, provide the health care that Australians want and need at prices they can afford to pay? If this is to be sustainable in the private sector health funds need to be given a capacity to manage their risk exposure. Provided insurers continue to insure sick people, costs will be driven not so much by the age profile of the insured population (though it will have some effect) as by technology and its capacity to supply and create the demands of the aged (not ageing per se). Handling these costs is the real challenge for the future. PMID- 12536863 TI - Informing policy and service development at the interfaces between acute and aged care. AB - This paper argues that policies to address the interfaces between acute care and aged care should view older people as members of the wider Australian population entitled to a range of health services under Medicare rather than focusing only on supposed "bed blockers". In seeking to explain the current level of policy interest in this area, three areas are canvassed: pressures on acute hospital care, particularly those attributed to population ageing; shrinking provision of residential aged care; and the proliferation of post acute services. If policy development is to maintain a wider rather than narrower perspective, attention needs to be given to improving collection and analysis of critical data that are currently unavailable, to developing system-wide funding arrangements for post acute care, and to reassessing what constitutes appropriate hospital activity for younger and older age groups alike. PMID- 12536864 TI - Has the increase in private health insurance uptake affected the Victorian public hospital surgical waiting list? AB - It was anticipated that increase uptake of Private Health Insurance (PHI) would reduce demand on public sector surgical waiting lists. The best measure of changed demand is the comparison of the actual cases added to that projected given previous trends in PHI uptake. Detailed Victorian data is available up to 2000-1. The total waiting list has varied little, reflecting significant decreases in both in patients added to and removed. There was a marked increase in private sector elective surgery cases coinciding with the fall in additions to the public sector waiting list and in public sector elective surgical cases. The June 2001 Victorian surgical waiting list would have been 69,599 not 41,838 if the PHI uptake rate had continued to fall in line with pre-1999 trends, and that of June 2002 about 100,000 compared to 40,458 in March 2002. Limited data from other states suggests the Victorian trends are representative of all Australia. PMID- 12536865 TI - The lore about private health insurance and pressure on public hospitals. PMID- 12536866 TI - Waiting time information services: how well do different statistics forecast a patient's wait? AB - This study investigates how accurately the waiting times of patients about to join a waiting list are predicted by the types of statistics disseminated via web based waiting time information services. Data were collected at a public hospital in Sydney, Australia, on elective surgery activity and waiting list behaviour from July 1995 to June 1998. The data covered 46 surgeons in 10 surgical specialties. The accuracy of the tested statistics varied greatly, being affected more by the characteristics and behaviour of a surgeon's waiting list than by how the statistics were derived. For those surgeons whose waiting times were often over six months, commonly used statistics can be very poor at forecasting patient waiting times. PMID- 12536867 TI - The significant impacts of workplace change on medical scientists in Victoria. AB - Pathology in Victoria has undergone considerable change in the last decade. The need for productivity gains, the effects of downsizing and budget cuts, alongside the effects of casemix funding, privatisation and the advent of Enterprise Bargaining Agreements (EBAs) have all placed considerable strain on medical scientists. This paper outlines the impacts of workplace change on medical scientists in Victoria, the most significant workplace stressors on medical scientists today, and what medical scientists feel needs to happen to enable them to regain control over their working lives. The findings from key informant interviews and focus groups of medical scientists show that radical changes in service requirements have occurred. The restructuring of laboratories to 'core' business and subsequent changes in client expectations has impacted on the standards of service delivery in pathology. These changes have resulted in considerable stress as the scientists struggle to cope with work intensification, increases in rostered work during 'unsociable' hours, the lack of control over change, and challenges to their professionalism as they feel that quality standards are not being met. PMID- 12536868 TI - Improving nursing recruitment and retention in a sub-acute health service. AB - The current worldwide shortage of nurses is well-documented. This paper describes one organisation's "back to basics" approach to nursing recruitment and the results of a vigorous recruitment program in 2001. Within 12 months, this organisation employed more than 200 new nursing staff, opened 67 beds, reduced agency use by 69% and reduced vacancies by 80%. Based on agency premium costs, there were minimum savings in the order of $800,000 per annum. PMID- 12536869 TI - Safe working hours--doctors in training a best practice issue. AB - In 1995, the Australian Medical Association launched its Safe Working Hours campaign. By 1998, this had been developed into a National Code of Conduct that continues to resonate in the Australian public health system. However, and particularly in respect of Doctors in Training (DITs) who continue to work long hours, there are levels of resistance to proposals that seek to re-organise work or change prevailing professional and cultural expectations. Long working hours have substantial impacts on a DIT's capacity to consistently deliver high quality patient care, dilute the effectiveness of their training regime and have negative consequences on their health, social life and family responsibilities. While public hospitals often maintain the view that minimal budget flexibility restricts their capacity to affect change in a positive way, in fact devisable productivity and efficiency gains can be achieved by reducing working hours. Further, the medical profession needs to consider whether long hours provide an optimal environment for quality learning and performance. PMID- 12536870 TI - Factors associated with young adults' obtaining general practitioner services. AB - This paper explores factors associated with the use of general practitioner services by young adults aged 20 to 24 years. The analyses examined sociodemographic and health data obtained for a community survey of respondents who lived in Canberra or Queanbeyan in 1999 and 2000 and linked this information to Medicare records of general practitioner visits over the following six months. In line with the Andersen model of health behaviour, predictor variables were classified as measures of physical and mental health needs for such services or as factors that predispose or enable the individual to obtain those services. Those who obtained general practitioner care were more likely to be female, to have been or be undertaking higher education and to be living with children. They were also identified by their having poorer physical health, more chronic diseases, higher levels of suicidal ideation and by their regularly using marijuana. General practitioners need to be aware of the potential mental health and substance use problems experienced by consumers in this age group. Strategies are also needed to encourage young men to seek general practitioner services when they are ill. PMID- 12536871 TI - Variation in levels of uptake of enhanced primary care item numbers between rural and urban settings, November 1999 to October 2001. AB - We aimed to report on variation in levels of uptake of enhanced primary care item numbers between rural and urban Divisions of General Practice between November 1999 and October 2001. Most providers of EPC services and most services (close to 70%) are located in capital cities and other metropolitan centres. The average number of health assessments done per provider was slightly lower (8-14) in remote than urban and rural (20-30) areas. A similar pattern was observed for care plans, but rates of case conferences were similar in rural and urban areas. However, adjusted for population aged 75 years and over, in all jurisdictions except South Australia, between 30% and 144% more health assessments were done per full time equivalent general practitioner (FTE GP) in rural divisions. For rural and urban Divisions of General Practice, there is a wide range in the rate of services provided, between and within Divisions. However, overall, more services are provided per FTE GP in rural Divisions. PMID- 12536872 TI - Video-conferencing: under-used by rural general practitioners. AB - The objective was to investigate the use of and value General Practitioners place on video-conferencing as a tool in providing rural health care. The participants were 8 rural general practitioners in rural Victoria towns. I found that six out of the eight GPs did not value video-conferencing as a tool to assist with patient care, and the other two GPs were interested in the technology only for certain aspects of support with patient consultations and continuing education. In conclude that there needs to be a review of whether video-conferencing equipment should continue to be implemented in the same way that it has been so far in Victoria, and of the cost-effectiveness of providing video-conferencing facilities in rural health services. In particular, there needs to be a review of whether more training and support for rural general practitioners is needed to increase the uptake of video-conferencing. Alternatively, analysis can be undertaken of the intrinsic value of using video-conferencing as an interactive tool for obtaining specialist support for patient care or undertaking continuing education via video-conferencing, and the program discontinued if it is found to be unwarranted. PMID- 12536873 TI - The Well Person's Health Check: a population screening program in indigenous communities in north Queensland. PMID- 12536874 TI - A decade of change in a community health service: a shift to acute and short-term care. AB - There have been major policy changes impacting on community health services such as early discharge and hospital in the home initiatives. The effects of such policies on the client profile of community health services are not well known. Administrative data was used to explore a decade of changes in South Western Sydney Area Health Service community health services. There is a clear shift from a service primarily focused on child health screening to one that is increasingly providing acute and shorter-term care to adults with physical health problems. The changes are reflective of policies that are moving acute treatment services into the community. PMID- 12536875 TI - Reorganisation of an ailing aged care service. AB - This paper evaluates the impact of reorganisation of geriatric and general medical services in a tertiary referral hospital. The patients involved were predominantly elderly with multiple medical problems. We used an iterative process of bed utilisation review, stakeholder consultation and service remodelling to improve bed management. Reduced length of stay and increased throughput allowed closure of between 22 and 30 beds and generated recurrent cost savings of $900,000 annually. Even under pressure it is possible to describe and evaluate the process of change. Such description and evaluation may help to avoid expensive mistakes. PMID- 12536876 TI - Unassigned Geriatric Evaluation and Management program: preventing sub-acute hospital admissions. AB - The purpose of this study was to describe and evaluate the Unassigned Geriatric Evaluation and Management (UGEM) program recently developed at Bundoora Extended Care Centre (BECC). The UGEM program resembles a bed substitution service and aims to prevent hospital admissions by providing community case management services to clients who would otherwise require inpatient admission. Data was collected on 36 clients who had received UGEM services. Twenty-six clients/carers also took part in a follow-up telephone satisfaction interview. Overall, the evaluation indicates that the UGEM program is a flexible, valuable and effective service. Most UGEM clients could be maintained at home with services, and this suggests that the program succeeds in assisting many clients to avoid hospitalisation in crisis situations. Further, clients and carers displayed a very high level of satisfaction with the service. The ongoing funding of such program is crucial in order to maintain the reduction in hospital admissions and hence the high costs associated with inpatient care. PMID- 12536877 TI - Treating organisational illness: a practical approach to facilitating improvements in health care. AB - The health sector contains many problems that are widely recognised and ought to be easily resolved, and yet some organisations seem to be powerless to act. We argue that this mainly reflects weaknesses in the organisational culture, and present an approach that we have been using to address them. We describe some simple analytical tools, and report our experiences in using them in organisations in several countries. We conclude that most people believe organisational weaknesses are important, are willing and eager to try to address them, and do in fact find ways of making some useful changes--at least, in the short-term. PMID- 12536878 TI - Organisational culture and safety: an interdependent relationship. AB - Since the early 1990s, a body of evidence regarding the lack of quality in health care has emerged in many countries including Australia, the United Kingdom, New Zealand and the United States of America. It has brought the subject of health care safety to the top of the policy agenda and the forefront of the public debate worldwide. Studies show not only that failure of quality occurs, but also that it inflicts harm and wastes resources on a large scale. Experts in risk management, both within and outside the health care industry, emphasize system failures and system-driven errors over direct human error, and accentuate the crucial role that organisational culture plays in ensuring safety. Examination of the interrelationship between culture and safety in organisations demonstrates that organisational relationships influence both culture and safety and that effective two-way communication is pivotal to the success of the development of a corporate 'safety culture'. PMID- 12536879 TI - Developing a positive organisational culture using a management development strategy. AB - This paper evaluates a management development program, which was piloted at Albury Base Hospital during 2001 for nurse managers. The program uses quantitative assessments of participants using a lifestyle inventory tool and enables managers to identify opportunities and strategies to assist them to become more effective managers. The program also helps identify management "blind spots" and increased awareness of the effect of management's behaviour on team efficiency. The program was evaluated as being relevant to managers from any health discipline and was a means by which workplace culture and behaviour could be improved. PMID- 12536880 TI - Improving the quality of hospital services: how diverse groups of consumers prefer to be involved. AB - Consumer participation in hospitals is increasingly being advocated for as a way to contribute to improving safety and quality. This article reports on the results of a study conducted to investigate if, and how, a range of consumers would prefer to be involved in improving the quality of hospital services. Face to-face interviews were conducted with 100 users of a hospital, telephone interviews were conducted with 2,005 members of the broader South Australian community, focus groups were conducted with 22 representatives of consumer and community groups and in-depth interviews conducted with four representatives who could not attend the focus groups. The results indicate that for users of the hospital and members of the broader community, there is a significant preference to be involved in more passive methods of participation that are initiated by the hospital, such as written and phone surveys. This is preferable to writing letters of complaint or compliment, participating in more active methods that require one-off commitment (focus groups and public planning forums) and ongoing commitment such as being members of working groups to address specific issues and hospital committees. In contrast, representatives of consumer and community groups have a strong preference to be involved in active methods of participation such as working groups to address specific issues and hospital committees. We found that there are considerable differences between age groups and educational attainment and preference for participation. If these issues are not taken into account, and a range of participation methods used, then there is the potential for some population groups to be excluded from having some input into improving the safety and quality of hospital services. PMID- 12536881 TI - The association between musculoskeletal disorders and obesity. AB - The aim of this study was to examine the association between musculoskeletal disorders and the level of obesity (as defined by the body mass index) for a sample of the Australian population aged 20-64. A logistic regression model was used to estimate the association between musculoskeletal disorders and obesity, controlling for a range of socio-demographic characteristics. Individual-level data on obesity, musculoskeletal disorders, and various socio-demographic characteristics were extracted from the Australian Bureau of Statistics (ABS) 1995 National Health Survey (NHS). Individuals with musculoskeletal disorders were identified using ICD-9 codes 710-739 from a sample of 28,376 individuals from the non-institutionalised population. Estimates from the logistic regression equation indicate that there is a statistically significant positive relationship between the probability of having a musculoskeletal disorder and the level of obesity. Socio-demographic variables such as age, sex, origin, income level, employment status and geographic location also had a statistically significant relationship. This information can be used by public health practitioners and educators to identify those at risk and to design health strategies that target at-risk patients. PMID- 12536882 TI - Cardiac care and surgery: future trends for America's #1 center of excellence. PMID- 12536883 TI - Trends in cardiac care: great news for patients and suppliers but a mixed outlook for providers. PMID- 12536884 TI - Market briefing: 'dollars and stents': new technology will have major impact on heart care. PMID- 12536885 TI - Medicare outpatient payment rates to rise almost 6 percent in 2003. PMID- 12536886 TI - Six steps to better labor management. PMID- 12536887 TI - Developing a plan to improve the odds of retaining your staff. PMID- 12536888 TI - What you as a manager can do to overcome verbal abuse of staff. AB - Employees and their work performance determine the success of any business. Permitting verbal abuse and other disruptive behaviors is bad management. Well written policies, physician champions, strong executives, and the willingness of OR managers to try new approaches to assist the staff to develop new skill sets will help to create healthier workplaces where everyone knows words don't hurt. PMID- 12536889 TI - Soap, alcohol rub OK for surgical scrub. PMID- 12536890 TI - Outpatient payments to rise by 3.7% in 2003. PMID- 12536891 TI - Expect big changes in JCAHO survey process. PMID- 12536892 TI - Borrowing from Toyota to improve quality. PMID- 12536893 TI - Equipment planning for a new OR suite. PMID- 12536894 TI - What's the best way to reward staff? PMID- 12536895 TI - Helping patients prepare for surgery. PMID- 12536896 TI - Perspectives. Thompson goes on road to seek ideas, support for demos. PMID- 12536897 TI - Between the lines. PMID- 12536898 TI - A piece of the action. PMID- 12536899 TI - There's the rub. PMID- 12536900 TI - Outpatient clinics. Down the line. AB - A hospital with a 157-week wait for orthopaedic outpatient appointments has introduced a telephone booking system which has reduced waiting by 53 weeks. The system, introduced with a 23,000 Pounds grant from the local health group, has cut non-attendance from 17 to 5 per cent and is estimated to have saved the orthopaedics department 216,000 Pounds. Two part-time clerical workers staff the initiative. Telephone booking is now being rolled out to include outpatient clinics of other specialties and orthopaedic day case lists. PMID- 12536901 TI - Hospital staffing. Vacant possession. AB - A review of nursing at an acute London trust with a deficit of 5 m Pounds has halved the vacancy rate and saved 500,000 Pounds a year on agency staff. The review took the director of nursing 170 hours. It has cut nurse turnover by over one-third. Once made aware of the costs of employing agency staff, nurses were encouraged to join the hospital bank. The trust has moved from from a one-star rating to three stars. PMID- 12536902 TI - Primary care. Home truths. AB - Charging social services for delays in patients being discharged from hospital is unlikely to result in improved care for older people. A new kind of community support worker dedicated to care of the elderly is needed. These workers could provide sustained support and prevent avoidable admissions to residential care. Care packages should take more account of what service users want. Government focus needs to shift from the management of beds to community resources. PMID- 12536903 TI - [Epidemiology. Status of bladder cancer in the world in the year 2000]. PMID- 12536904 TI - [Diagnosis of infiltrating tumor of the bladder]. PMID- 12536905 TI - [Definition and classification of infiltrating tumors of the bladder]. PMID- 12536906 TI - [Management of a cystectomy specimen: total cysto-prostatectomy or anterior pelvic surgery]. PMID- 12536907 TI - [Pathologic anatomy of infiltrating epithelial tumors of the bladder]. PMID- 12536908 TI - [Histologic prognosis of infiltrating urothelial tumors]. PMID- 12536909 TI - [Pathologic anatomy of infiltrating non-epithelial tumors of the bladder]. PMID- 12536910 TI - [Pre-therapeutic evaluation of infiltrating tumors of the bladder]. PMID- 12536911 TI - [Total cystectomy: surgical techniques]. PMID- 12536912 TI - [Urinary diversion: surgical techniques]. PMID- 12536913 TI - [Mortality, morbidity, and functional results of total cystectomy]. PMID- 12536914 TI - [Results of cystectomy for cancer]. PMID- 12536915 TI - [Prostatic invasion and indications for urethra surgery]. PMID- 12536916 TI - [pN+ lymphatic invasion and total cystectomy]. PMID- 12536917 TI - [Analysis of total cystectomy in elderly patients]. PMID- 12536918 TI - [Quality of life and urinary diversion]. PMID- 12536919 TI - [Prognostic factors of survival after total cystectomy]. PMID- 12536920 TI - [Fortuitous disclosure of prostatic cancer after examination of the cystoprostatectomy specimen]. PMID- 12536921 TI - [Chemotherapy of locally advanced T4 and metastatic N+, M+ bladder tumors]. PMID- 12536922 TI - [Chemotherapy protocols: new perspectives]. PMID- 12536923 TI - [Neo-adjuvant and adjuvant chemotherapy in radical treatment]. PMID- 12536924 TI - [Metastatic or locally advanced tumors of the bladder: is there a role for adjuvant surgery in chemotherapy?]. PMID- 12536925 TI - [Transurethral resection of bladder only]. PMID- 12536926 TI - [External radiotherapy]. PMID- 12536927 TI - [Simultaneous combined radiotherapy and chemotherapy]. PMID- 12536928 TI - [Conservative alternatives of bladder reservoir with more unusual indications]. PMID- 12536929 TI - [Molecular markers of infiltrating cancer of the bladder]. PMID- 12536930 TI - [Tumor recurrence in the upper urinary tract after total cystectomy]. PMID- 12536931 TI - [Tumor recurrence in male anterior urethra after total cystectomy]. PMID- 12536932 TI - [Pelvic tumor recurrence after total cystectomy]. PMID- 12536933 TI - [Intra-diverticular urothelial tumors of the bladder]. PMID- 12536934 TI - [Epidermoid carcinoma of the bladder]. PMID- 12536935 TI - [Adenocarcinoma of the bladder]. PMID- 12536936 TI - [Small cell and/or neuroendocrine cell cancer of the bladder]. PMID- 12536937 TI - [Sarcomatoid carcinoma]. PMID- 12536939 TI - [Synthesis on the treatments of infiltrating tumors of the bladder]. PMID- 12536938 TI - [Post-therapeutic surveillance of infiltrating tumors of the bladder]. PMID- 12536940 TI - [AFU recommendations 1998. "Committee on Cancer of the French Association of Urology"]. PMID- 12536941 TI - [EAU Recommendations 2001. "Guidelines on bladder cancer"]. PMID- 12536942 TI - [Bladder cancer. Clinical cases]. PMID- 12536943 TI - Religious identity and the health care market: mergers and acquisitions involving religiously affiliated providers. PMID- 12536944 TI - Engelhardt, the IK, and the Foundations of Bioethics. PMID- 12536945 TI - Bioethics in an old key. PMID- 12536946 TI - Of intellectual history, postmodern ethical banality, and the search for moral content. PMID- 12536947 TI - Scofield's misdiagnosis of Engelhardt's Foundations of Christian Bioethics. PMID- 12536948 TI - After voids: Scofield vs. Engelhardt. PMID- 12536949 TI - Eyes wide shut: Scofield on Engelhardt. PMID- 12536950 TI - Philosophy vs. religion in bioethics: Scofield vs. Engelhardt. PMID- 12536951 TI - Taking the lead for accountability. PMID- 12536952 TI - Don't read too much into Oregon single-payer vote. PMID- 12536953 TI - Cashing in on obesity? PMID- 12536954 TI - Providers turn to fee-for-service charges to make up revenue lost under capitation. PMID- 12536955 TI - Blues on the move. PMID- 12536956 TI - Beware the hidden consequences of the malpractice crisis. PMID- 12536957 TI - Placing faith in technology to improve members' choices. Interview by Patrick Mullen. PMID- 12536958 TI - Advisory boards create company-plan cooperation. PMID- 12536959 TI - Same-day appointments promise increased productivity. PMID- 12536960 TI - Modified HIPAA privacy rule affects research, marketing, security. PMID- 12536961 TI - Providers, plans misinformed about vendor software capability. PMID- 12536962 TI - M+C meltdown hinders access to medications. PMID- 12536963 TI - [Evaluation of hearing thresholds of 40Hz auditory event related potential and auditory brainstem response]. AB - Recordings of pure tone response, 40 Hz auditory event-related potential (40 Hz AERP) and auditory brainstem response (ABR) were obtained for 74 ears of 42 cases (32 normal ears, 42 injured ears). 40 Hz AERP (0.5-2 kHz) of 20 ears was tested under natural sleep and awake conditions. The results showed that the threshold of 40 Hz AERP was higher than that of tone pip, with the difference of 12.7 +/- 6.4 dBnHL at 0.5 kHz, 14.7 +/- 6.3 dBnHL at 1 kHz, and 15.6 +/- 5.6 dBnHL at 2 kHz respectively. The threshold of 40 Hz AERP increased in natural sleep compared with that in awake state. The threshold of ABR was higher than the behavioral. The data suggests that jointly using several tests get more accurate and objective results in evaluating hearing loss. PMID- 12536964 TI - [Motor evoked potentials of upper limbs]. AB - This study aims to explore a method to record and analyze motor evoked potentials (MEP) of upper limbs. Compound muscle action potentials (CMAP) are recorded from abductor digiti minimi (ADM) and abductor pollicis brevis (APB) by magnetic stimulation of human brain and peripheral nervous. Results show that the latency of left and right Cz-APB is 25.23 ms and 25.19 ms respectively. The difference of the bilateral latencies is 2.24 ms. The rate of amplitude, the rate of area for bilateral sides is 0.45 and 0.37. The left and right central conduction time (CCT) of Cz-APB is 11.79 ms and 11.75 ms. The bilateral difference is 2.00 ms. The latency of left and right Cz-ADM is 25.23 ms and 25.19 ms respectively. The difference of bilateral latencies is 2.24 ms. The rate of amplitude, the rate of area for bilateral sides is 0.45 and 0.37. The left and right CCT of Cz-ADM is 11.79 ms and 11.75 ms. The bilateral difference is 1.86 ms. PMID- 12536965 TI - [Development and application of the software measuring the size of injured body surface]. AB - A software to measure the area, length and angle of injured body surface is introduced. It is suitable for evaluating injury degrees with 2 or 3 dimension measurement. The software is developed by Visual Basic 6.0 on Windows 98 terrace, supporting digital video camera, digital camera and scanner input. It is easy to use and has been applied in more than 200 cases. PMID- 12536966 TI - [Immunohistochemical study of basic fibroblast growth factor of early myocardial infarction]. AB - The expression of basic fibroblast growth factor (bFGF) was first studied in 61 cases of early myocardial infarction. The results revealed that all 24 cases of proven infarcts was positive in nuclei of myocardium and vascular endothelium, and that some was positive in cytoplasm, especially in myocardium around the border zone of infarction. Of 21 suspected infarcts cases, 17 showed strong positive and 4 mild positive. In the control group of 16 samples, only 3 was mild positive sparsely in nuclei of myocardium and vascular endothelium. PI of control group was significantly lower than that of proven and suspected infarcts (P < 0.01). There was no significant difference between PI of proven infarcts and suspected infarcts (P > 0.05). These findings suggest that SABC-bFGF method could be used for morphologic diagnosis of early myocardial infarction of sudden cardiac death. PMID- 12536967 TI - [Expression of TGF-beta 1 and proliferation of collagen in myocardium in viral myocarditis]. AB - To investigate the tissue repair of acute viral myocarditis, and to explore the diagnostic method of slight viral myocarditis in forensic pathology. Slight viral myocarditis model was induced in Balb/c murine by CVB3. Collagen proliferation in myocardium of mice with myocarditis was observed by special staining. The hearts of mice and human(9 cases) with myocarditis were studied LSAB immunohistochemically with anti-TGF-beta 1 antibody. In the study, the proliferation of collagen was seen in myocardium in acute viral myocarditis. Generous expression of TGF-beta 1 was found in the myocardium of mice and human with myocarditis. The quantity of collagen proliferation and expression of TGF beta 1 was positive correlation. It is concluded that the tissue repair exists in acute viral myocarditis and that positive staining of myocardium for TGF-beta 1 is a sensitive index of myocardial damage and tissue repair. PMID- 12536968 TI - [Experimental study on basic fibroblast growth factors in viral myocarditis]. AB - In order to know earlier pathological changes in myocarditis, 8 rats were inoculated with CVB3, and a model of viral myocarditis was built. The changes of basic fibroblast growth factors (bFGF) in acute viral myocarditis were observed by immunohistochemistry. It was found that the bFGF stain started to show positive at the third day after viral infection and that the positive degree increased afterwards. With normal HE staining, obvious lesion usually appeared under microscopy at least five days after infection. This suggests that the changes of bFGF in myocardium can be used for examination of early and light lesions of viral myocarditis. PMID- 12536969 TI - [Study on histology and the expression of Bax and Bcl-2 after cerebral contusion in rats]. AB - To set up the experimental model of focal cerebral contusions in rats, and to study the histology and the expression of Bax and Bcl-2 after cerebral catusions. It was found that neurons and astrocytes around the wound area were changed morphologically and that polymorphonuclear leucocytes were present at 12-24 h after injury. A lot of foamy cells and astrocytes having a big dark nucleus appeared around the wound area on the 4th day after injury. On the 8-10th day, liquefaction necrosis was seen in areas of contusions. On the 12-14th day, the contusion focus healed up and became cystiform or glial node. Meantime, lots of astrocytes, capillary vessel and hemosiderin were observed. The expression of Bax and Bcl-2 increased after injury. There existed a relationship between increased Bax/Bcl-2 and different intervals after brain injury. PMID- 12536970 TI - [Usefulness of Cofiler PCR amplification kit in parentage testing]. AB - Parentage testing was performed by using multiple PCR and automated genescan and genotype analysis. Parentage was excluded in 36 cases, including 22 of 149 paternity trios and 14 of 37 paternity duos. In the 22 trios and 14 duos in which paternity was excluded, a mean of 6.1 and 5 incompatible STR markers were noted. The application value of six loci which included in the Cofiler amplification kit was showed here. The discrimination power ranged from 0.8023 for TPOX to 0.9203 for D16S539. The cumulative power of exclusion was provided as 0.9825. Three mutation were found (one each at D3S1358, TPOX, CSF1PO). For most nonexcluded paternity duos, only when the Cofiler amplification kit was used in conjunction with Profiler Plus kit, the RCP value was acceptable. The data showed that the Cofiler kit could play an important role in cases of paternity duos. PMID- 12536971 TI - [Evaluation of 9 STR loci in paternity identification]. AB - 9 STR loci obtained by four-dye fluorescent labeling technique in paternity identification provides much information at one test and the cumulative chance of exclusion gets up to 0.9999. Our result of 268 paternity test cases shows that there are at least two incompatible loci in all Mother-Child-Alleged Father (M-C AF) exclusive cases. To those unexclusive cases, The RCP all reaches international standard. It is suggested that more STR loci be used for accurate test in Child-Alleged Father(C-AF) case. PMID- 12536972 TI - Induction of syngeneic intradermal and orthotopic epidermal carcinomas in hairless Skh-1 mice as a reproducible model for experiments. AB - Epidermoid carcinomas, clinically and histologically similar to human squamous cell carcinomas (SCC), were obtained in hairless Skh-1 mice. Tumor cells originated from chemically-induced skin cancers. We developed three models of orthotopic skin tumors: (1) intradermal injection of a tumor cell suspension, (2) superficial abrasion of the skin, cell grafting and application of a hydrocolloid dressing, (3) skin incision, seeding and application of a hydrocolloid dressing. Intradermal injection was 100% successful. Skin incision, displaying histological evidence of rapid invasive tumor growth, was 75% successful. Though skin tumor growth after abrasion was only 20% successful, the tumor histogenesis exactly imitated human SCC development. These carcinomas provide research models for further experiments such as photodynamic therapy or antiangiogenesis therapy. PMID- 12536973 TI - [Assessment of single-stranded DNA breaks in spleen lymphocytes using the DNA comet assay in the long term after acute irradiation of mice at sublethal and moderately-lethal doses]. AB - The single-strand DNA level in spleen lymphocytes of BALB/c male-mice after 11 month acute exposure to gamma-radiation at doses 1, 3 and 6 Gy has been investigated by comet assay. The results of our study showed that at 11 month after irradiation at different doses a significant increase in the level of DNA breaks in spleen lymphocytes and decrease in the total number of these cells in mice was registered. It is possible that the increase in the DNA breaks is due to the effort of the compensatory proliferation process in lymphoid system that can give the increase in the number of different genetic disturbances in lymphocytes. PMID- 12536975 TI - Medical school applications. Compensating factors do not compensate for poor A level performance. PMID- 12536976 TI - Medical school applications. Applications have increased again for second year running. PMID- 12536978 TI - Public access defibrillators. Defibrillators in public places save lives. PMID- 12536979 TI - Public access defibrillators. Fire extinguisher model of public access defibrillation is unlikely to prevent many deaths. PMID- 12536980 TI - Public access defibrillators. Data from Cuba. PMID- 12536981 TI - Warfarin and extractions. PMID- 12536982 TI - Shock waves. PMID- 12536983 TI - Premolar restoration. PMID- 12536984 TI - Theme issue for medics and health informed public. What the future might hold for the BMJ in 2013. PMID- 12536985 TI - Antibiotic prophylaxis. PMID- 12536986 TI - Professional opinion. PMID- 12536987 TI - So, you'd like a handheld computer. PMID- 12536988 TI - The Erice Statement on drug advertising to consumers. PMID- 12536989 TI - CYP2D6 extensive, intermediate, and poor phenotypes and genotypes in a Polish population. AB - OBJECTIVE: This study investigated the distribution of the CYP2D6 genotypes and phenotypes in a Polish population and compared the concordance of the two methods. METHODS: Six hundred unrelated healthy individuals from southwestern Poland were studied. The CYP2D6 phenotype was analyzed in 300 individuals using sparteine as a model drug. The CYP2D6 genotype was analyzed in 300 individuals by polymerase chain reaction amplification and restriction fragment length polymorphism techniques for the CYP2D6*1, CYP2D6*3, and CYP2D6*4 alleles. Additionally, in 60 randomly selected healthy individuals both the CYP2D6 phenotype and genotype was assessed to determine accordance between the methods. RESULTS: Of 300 participants in the study 25 (8.3%) were classified as poor metabolizers, 44 (14.7%) as intermediate metabolizers, and 231 (77%) as extensive metabolizers of sparteine. The frequency of CYP2D6*1, CYP2D6*3, and CYP2D6*4 alleles among the genotyped 300 persons was 75.7%, 1.3%, and 23.0%, respectively. The frequency of CYP2D6 deficient genotypes in a Polish population (8.0%) was similar to phenotyping results. The comparison of phenotype and genotype in 60 randomly selected individuals showed a good concordance of the obtained results. CONCLUSIONS. The frequencies of poor metabolizers for CYP2D6 phenotype (8.3%) and genotype (8.0%) in a Polish population from the southwestern region are in concordance and compare well with most results of poor oxidation metabolizers in other white populations. PMID- 12536990 TI - Mulitvariate cluster analysis of a human hepatic cytochrome P450 database. PMID- 12536991 TI - Successful treatment of multi-resistant Stenotrophomonas maltophilia meningitis with ciprofloxacin in a pre-term infant. PMID- 12536992 TI - How frequent is coeliac disease in Down syndrome? PMID- 12536993 TI - Macrophage activation syndrome after autologous stem cell transplantation for systemic juvenile idiopathic arthritis. PMID- 12536994 TI - The impact of the control of serum phenylalanine levels on osteopenia in patients with phenylketonuria. PMID- 12536995 TI - Acute neonatal collapse resulting from pericardial effusion. PMID- 12536996 TI - Use of unconventional medicine in children in Italy. PMID- 12536998 TI - Bibliography of toxinology. PMID- 12536997 TI - The European Registry of Cardiac Catheter Interventions 1997. PMID- 12536999 TI - You're the flight surgeon. Acute diverticulitis. PMID- 12537000 TI - Current status of sexing mammalian spermatozoa. AB - Thousands of offspring have now been produced via artificial insemination with spermatozoa sexed by flow cytometry and cell sorting. We are unaware of any other practical approach to sexing spermatozoa that maintains fertility. Accuracy of sexing usually is 85-95% in most species, but somewhat lower with human spermatozoa. Spermatozoa are sexed in series, one at a time, at routine rates of about 3000 live spermatozoa of each sex per second for most species, and nearly twice that rate under optimal conditions for some species. Owing to various constraints and statistical considerations, there appears to be an upper theoretical limit to sexing spermatozoa of about 10,000 live spermatozoa of each sex per second with current methodology. About a quarter of the spermatozoa processed are sexed; the rest are discarded in the process or lost due to logistical constraints. Spermatozoa undergo some damage during sorting, although much less in terms of viability than with routine cryopreservation; fertility is lower with sexed than control spermatozoa. Offspring from sexed spermatozoa appear to have no more abnormalities than do controls, and both groups grow and thrive similarly. Despite high costs and complex procedures, sexing spermatozoa, usually followed by cryopreservation, is being used commercially for cattle and horse production in several countries, and is used to produce girls to avoid X chromosome-linked genetic diseases. PMID- 12537001 TI - Cutaneous reaction to oral spironolactone with positive patch test. PMID- 12537002 TI - Nuclear medicine asleep in sleep research? PMID- 12537003 TI - 90Y-DOTATOC and nephrotoxicity. PMID- 12537004 TI - Troubling lesion subsets: bifurcation disease. PMID- 12537005 TI - Landmarks: examining the foundation for GP IIb/IIIa inhibitors in PCI. PMID- 12537006 TI - Is sentinel lymph node investigation useful for early tumour stages only? PMID- 12537007 TI - Occupancy of dopamine D2 receptors in the mouse brain measured using ultra-high resolution single-photon emission tomography and [123]IBF. AB - Functional imaging of small animals, such as mice and rats, using ultra-high resolution positron emission tomography (PET) and single-photon emission tomography (SPET) should be a valuable tool in studies of drug occupancy of cerebral binding sites. In this study we aimed to demonstrate the feasibility of using ultra-high-resolution SPET to measure the occupancy of dopamine D2 receptors by a competing drug, using the dopamine D2 receptor-specific radioligand iodine-123 5-iodo-7-N-[(1-ethyl-2-pyrrolidinyl) methyl] carboxamido 2,3-dihydrobenzofuran ([123I]IBF). Fourteen normal male mice (CD-1) were jugular vein-cannulated and a bolus infusion protocol was used to deliver 360 MBq [123I]IBF into the mouse (bolus-to-infusion ratio 1.8:1). The mice were scanned using an ultra-high-resolution triple-headed SPET system equipped with pinhole collimators. After sustained equilibrium had been achieved, varying doses of raclopride, a potent dopamine D2 receptor antagonist, were injected through the tail vein and the tracer was allowed to regain equilibrium. A simple equilibrium ratio of striatum to cerebellum provided a measure of D2 receptor binding both before and after injection of raclopride. Following raclopride administration, the system returned to equilibrium with lower specific binding in the striatum, while the counts in the cerebellum were unaffected. Receptor occupancy was 5.2% +/- 2.9% (control), 52.1% +/- 11.1% (0.3 mg/kg), 79.3% +/- 4.8% (1.0 mg/kg), and 94.7% +/- 2.2% (3.0 mg/kg), which gave an ED50=0.26 +/- 0.03 mg/kg using a single receptor site saturation model. This study has demonstrated clearly that ultra high-resolution SPET of small animals is capable of measuring displacement and occupancy of dopamine D2 receptors by competing ligands. PMID- 12537009 TI - For: Is LSO the future of PET? PMID- 12537008 TI - Effect of angiotensin converting enzyme inhibition on myocardial phosphoinositide metabolism visualised with 1-[1-11C]-butyryl-2-palmitoyl-rac-glycerol in myocardial infarction in the rat. AB - We recently reported that myocardial phosphoinositide (PI) metabolism can be visualised by 1-[1-11C]-butyryl-2-palmitoyl-rac-glycerol (11C-DAG) in rats with myocardial infarction (MI). Angiotensin II, the receptors for which are expressed predominantly in infarcted areas with active fibrogenesis rather than in non infarcted regions, is involved in the upstream signalling systems of PI metabolism and plays an important role in the process of left ventricular (LV) remodelling after MI. We therefore hypothesised that the distribution of 11C-DAG after MI may be affected by the inhibition of angiotensin converting enzyme, which is one of the most important factors in the development of LV remodelling after MI. Rats were injected with 11C-DAG after 3 or 10 weeks of treatment with captopril or no treatment following coronary artery ligation, and quantitative autoradiography was performed. Cells occupying the infarcted region were identified by immunohistochemistry. Compared with untreated rats, treatment with captopril for 3 weeks after MI elicited a reduction in the 11C-DAG uptake in the infarcted region (P<0.05) but not in the non-infarcted region, and was associated with a 22% decrease in the heart weight/body weight ratio. The thallium-201 distribution in the infarcted area was similarly low in the rats with and rats without the 3-week captopril treatment after MI. Abundant macrophages and myofibroblasts occupied the infarcted area in both rats with and rats without the captopril treatment for 3 weeks after MI. The 11C-DAG radioactivity in the infarcted region in the untreated rats was lower 10 weeks after MI than 3 weeks after MI (P<0.01). This finding was in agreement with the results of immunohistochemistry demonstrating that the number and size of macrophages and myofibroblasts were remarkably reduced in rats 10 weeks after MI compared with 3 weeks after MI. Captopril treatment for 10 weeks after MI did not decrease the 11C-DAG radioactivity in the infarcted area further. These data suggest that 11C DAG is useful for visually detecting regions with activated PI metabolism after MI, and that captopril reduces PI metabolism in the infarcted region in the relatively early phase of MI, which might contribute to the attenuation of ventricular remodelling. PMID- 12537010 TI - Against: Is LSO the future of PET? PMID- 12537011 TI - New North American research network focuses on biliary atresia and neonatal liver disease. PMID- 12537013 TI - Abstracts of the 10th German Mapping Meeting. Giessen, Germany. September 28-29, 2001. PMID- 12537012 TI - Gastric tonometry in the care of the newborn. PMID- 12537015 TI - Introducing the mouse--in As, Cs, Ts, and Gs. PMID- 12537016 TI - Dermatological infectiology. PMID- 12537017 TI - The synthesis of anti-fixed 3-methyl-3-deaza-2'-deoxyadenosine and other 3H imidazo[4,5-c]pyridine analogs. AB - Rotation of a heterocyclic base around a glycosidic bond allows the formation of syn and anti conformations in nucleosides. The syn conformation has been observed primarily in purine-purine mismatches in DNA duplexes. Such mismatches give rise to false positive oligonucleotide hybridization in DNA-based diagnostics. Here we describe the synthesis of an analog of 2'-deoxyadenosine that retains its Watson Crick functional groups, but cannot form the syn conformation. In this analog, the N3 atom of 2'-deoxyadenosine is replaced by a C-CH3 group to give 7-methyl-1 beta-D-deoxyribofuranosyl-1H-imidazo[4,5-c]pyridin-4-ylamine or 3-methyl-3-deaza 2'-deoxyadenosine (3mddA). This modification sterically prevents the syn conformation and 3mddA becomes an anti-fixed nucleoside analog of 2' deoxyadenosine. The synthesis and conformational analysis of 3mddA and several analogs with an 3H-imidazo[4,5-c]pyridine skeleton are described, as well as their potential applications. PMID- 12537018 TI - Affinity modification of EcoRII DNA methyltransferase by the dialdehyde substituted DNA duplexes: mapping the enzyme region that interacts with DNA. AB - Affinity modification of EcoRII DNA methyltransferase (M x EcoRII) by DNA duplexes containing oxidized 2'-O-beta-D-ribofuranosylcytidine (Crib*) or 1-(beta D-galactopyranosyl)thymine (Tgal*) residues was performed. Cross-linking yields do not change irrespective of whether active Crib* replaces an outer or an inner (target) deoxycytidine within the EcoRII recognition site. Chemical hydrolysis of M x EcoRII in the covalent cross-linked complex with the Tgal*-substituted DNA indicates the region Gly268-Met391 of the methylase that is likely to interact with the DNA sugar-phosphate backbone. Both specific and non-specific DNA interact with the same M x EcoRII region. Our results support the theoretically predicted DNA binding region of M x EcoRII. PMID- 12537019 TI - Synthesis of 2'-deoxy-2'-fluoroguanyl-(3',5')-guanosine. AB - The protected analogue of 2-amnio-6-chloropurine arabinoside (3b) was subjected to reaction with diethylaminosulfur trifluoride (DAST) and subsequently treated with NaOAc in Ac2O/AcOH to give N2, O3', O5'-triacetyl-2'-deoxy-2' fluoroguanosine (5a). After deacetylation of the sugar moiety and protection of 5'-OH by a 4,4'-dimethoxytrityl group, this nucleoside component was converted to 2'-deoxy-2'-fluoroguanyl-(3',5')-guanosine (6c, GfpG). PMID- 12537020 TI - Targeting of A701G nucleotide at the human ATP1A1 locus using a RNA/DNA chimera. AB - The single base substitution mediated by chimeric RNA/DNA oligonucleotide is a new promising approach of gene therapy for single base mutation diseases. We exploited this approach to render HeLa cells resistant to ouabain by introducing a single base substitution in the alpha 1 subunit of the NA+/K+ ATPase human gene. The chimeric oligonucleotide was administered to HeLa cells by electroporation and the frequency of ouabain resistant cells determined. The results showed that the chimeric RNA/DNA oligonucleotide failed to enhance the frequency of ouabain resistant cells supporting the controversy about the conflicting results of the technique. PMID- 12537021 TI - A comparison of the properties and the solution structure for RNA and DNA quadruplexes which contain two GGAGG sequences joined with a tetranucleotide linker. AB - We have determined solution structure of r(GGAGGUUUUGGAGG) (R14) by NMR; the RNA 14-mer forms an intra-strand parallel quadruplex with a G-tetrad and a hexad, in which a G-tetrad core is augmented by association of two A residues. The quadruplex further forms a dimer through stacking interaction between the hexads. In order to obtain insight into the difference between RNA and DNA quadruplexes, we synthesized the corresponding DNA 14-mer, d(GGAGGTTTTGGAGG) (D14), and examined its properties and structure by CD, gel electrophoresis, and NMR. K+ ions increased the thermal stability of both R14 and D14 structures. The binding affinity of K+ ions to R14 was much higher than that to D14. The CD and gel electrophoretic studies suggest that D14 forms a quadruplex entirely different from that of R14 in the presence of K+ ions; two molecules of D14 form a quadruplex with both antiparallel and parallel strand alignments and with diagonal loops at both ends of the stacked G-tetrads. The NMR study also gave results that are consistent with such structure: alternate glycosidic conformation, 5'G(syn)-G(anti)3', and characteristic chemical shift data observed for many quadruplexes containing diagonal TTTT loops. PMID- 12537022 TI - Procedure for selecting starting conformations for energy minimization of nucleosides and nucleotides. AB - The purpose of this study was to carry out a thorough search of the conformational space of various adenine-containing nucleotides, applying a previously published searching procedure, known as the representative method. This method, which reduces the number of starting conformations required to explore all the important regions of conformational space, appears to be successful in finding all (or nearly all) the putative low-energy conformations of each molecule. PMID- 12537023 TI - Kinetic and molecular modeling of nucleoside and nucleotide inhibition of malate dehydrogenase. AB - We studied the inhibition of mitochondrial malate dehydrogenase (mMDH) by the nucleotides cAMP, AMP, ADP, ATP. The experimental kinetic studies showed that the nucleotides were competitive inhibitors and that cAMP was probably the most potent inhibitor. To explain these observations, we used molecular modeling to determine the location, orientation, and relative binding energy of the nucleotides to mMDH. The order of the calculated binding energies, from lowest (most favorable) to highest, was cAMP, AMP, ADP, and ATP, which corresponded somewhat to the order of the experimentally determined inhibition constants. PMID- 12537024 TI - Synthesis and antiviral evaluation of isatin ribonucleosides. AB - A series of novel substituted isatin ribonucleosides 3b-3f were synthesized in good yields by a TMSOTf catalysed coupling reaction between the silylated nitrogenated base (1b-1f) and 1-O-acetyl-2,3,5-tri-O-benzoyl-beta-D-ribofuranose (2). Isatin nucleoside 3a previously reported was also prepared using this method giving high yield. From the compounds tested, ribonucleoside 3f proved to be the most active one when assayed for antiviral activitiy on HSV-1 infected cells, leading to 66% of inhibition of virus yield. All the isatin derivatives tested did not inhibit HIV-1 Reverse Transcriptase (RT) activity. PMID- 12537025 TI - A new class of biheterocyclic thioglycosides from pyridine-2-(1H)-thiones. AB - A reported method for preparation of a new class of biheterocyclic thioglycosides via reaction of pyridinethiones with 2,3,4,6-tetra-O-acetyl-alpha-D-gluco- and galactopyranosyl bromides has been studied. PMID- 12537026 TI - Convenient synthesis of fluorinated nucleosides with perfluoroalkanesulfonyl fluorides. AB - Perfluoroalkanesulfonyl fluorides are effective dehydroxy-fluorination agents for the hydroxyl group at the sugar moiety of nucleoside derivatives and give the corresponding fluorinated nucleosides in good yield with an inversion of configuration in a single step. PMID- 12537027 TI - Synthesis, structure, and conformation of anti-tumor agents in the solid and solution states: hydroxyl derivatives of Ftorafur. AB - The pyrimidine antimetabolite Ftorafur [FT; 5-fluoro-1-(tetrahydro-2 furyl)uracil] has shown significant antitumor activity in several adenocarcinomas with a spectrum of activity similar to, but less toxic than, 5-fluorouracil (5 FU). It is considered as a prodrug that acts as a depot form of 5-FU, and hence the two drugs exhibit a similar spectrum of chemotherapeutic activity. Ftorafur is metabolized in animals and humans when hydroxyl groups are introduced into the tetrahydrofuran moiety. These metabolites are also thought to be as active as ftorafur but less toxic than 5-FU. Hydroxyl derivatives: 2'-hydroxyftorafur (III), 3'-hydroxyftorafur (IV) and 2',3'-dihydroxyftorafur (II) were synthesized and X-ray and NMR studies of these hydroxyl derivatives were undertaken in our laboratories to study the structural and conformational features of Ftorafur and its metabolites in the solid and solution states. X-ray crystallographic investigations were carried out with data collected on a CAD-4 diffractometer. The structures were solved and refined using the SDP crystallographic package of Enraf-Nonius on PDP 11/34 and Microvax computers. All of the compounds studied had the base in the anti conformation. The glycosidic torsion angles varied from 20 to 60 degrees. There is an inverse correlation between the glycosyl bond distances and the chi angle. Molecules with a lower chi angle have a larger bond distance and vice versa. The sugar rings show a wide variation of conformations ranging from C2'-endo through C3'-endo to C4'-exo. The crystal structures are stabilized by hydrogen bonds involving the base nitrogen atom N3 and the hydroxyl oxygen atoms of the sugar rings as donors and the keto oxygens O2 and O4 of the base and the hydroxyl oxygen atoms O2' and O3' as acceptors. The NMR studies were carried out on Bruker 400 and 600 MHz instruments. Simulated proton spectra were obtained through Laocoon, and pseudorotational parameters were solved by Pseurot. Presence of syn or anti forms was demonstrated with the use of NOE experiments. The glycosyl conformations in solution vary more widely than in the solid state. The conformations of the sugar molecules are in agreement with the values obtained in the solid state. The studies of the structure and conformation in the solid and solution states give a model for the Ftorafur molecule that could be used in structure, function and biological activity correlation studies. PMID- 12537028 TI - Application of Mitsunobu reaction to solid-phase peptide nucleic acid (PNA) monomer synthesis. AB - PNA type I monomer backbone with a reduced peptide bond was synthesized on a Merrifield resin in Mitsunobu reaction of Boc-aminoethanol with resin-bound o nitrobenzenesulfonylglycine. The pseudodipeptide secondary amine group was deprotected by thiolysis and acylated with thymin-1-ylacetic acid. The monomer was released as a methyl ester. The procedure seems to be of general applicability and allows various modifications of PNA structure by using diverse alcohols and amino acid esters. PMID- 12537029 TI - Synthesis of novel 8-substituted carbocyclic analogs of 2',3'-dideoxyadenosine with activity against hepatitis B virus. AB - Synthesis and antiviral activity of several new 8-substituted carbocyclic analogs of D-2',3'-dideoxyadenosine are described. The new 8-substituted analogs were synthesized via lithiation of carbocyclic 2',3'-dideoxyadenosine followed by quenching with electrophiles. This methodology allows for a divergent synthesis of a variety of 8-substituted analogs from carbocyclic 2',3'-dideoxyadenosine in high yields. 8-Methyl and 8-halogenated carbocyclic 2',3'-dideoxyadenosine analogs showed 6-25 fold more activity against hepatitis B virus than the unsubstituted carbocyclic D-2',3'-dideoxyadenosine. PMID- 12537030 TI - The link between cannabis use and psychosis: furthering the debate. PMID- 12537031 TI - Focal brain stimulation with repetitive transcranial magnetic stimulation (rTMS): implications for the neural circuitry of depression. PMID- 12537032 TI - Cannabis dependence and psychotic symptoms in young people. AB - BACKGROUND: The aims of this research were to use data gathered over the course of a 21 year longitudinal study to examine the linkages between cannabis dependence at ages 18 and 21 and rates of psychotic symptoms taking into account previous symptom levels and other confounding factors. METHOD: Data were gathered during the course of the Christchurch Health and Development Study (CHDS). The CHDS is a longitudinal study of a birth cohort of 1,265 children who have been studied from birth to age 21. As part of this study, data were gathered on cannabis dependence and psychotic symptoms at ages 18 and 21. RESULTS: Young people meeting DSM-IV criteria for cannabis dependence had elevated rates of psychotic symptoms at ages 18 (rate ratio = 3.7; 95% CI 2.8-5.0; P < 0.0001) and 21 (rate ratio = 2.3; 95% CI 1.7-3.2; P < 0.0001). These associations were adjusted for previous psychotic symptoms and a range of other confounding factors using a generalized estimating equation model. This analysis showed that after adjustment for confounding factors, those meeting criteria for cannabis dependence still had an increased rate of psychotic symptoms (rate ratio = 1.8; 95 % CI 1.2-2.6; P < 0.005). CONCLUSIONS: The results show that the development of cannabis dependence is associated with increased rates of psychotic symptoms in young people even when pre-existing symptoms and other background factors are taken into account. PMID- 12537033 TI - Effects of cannabis and psychosis vulnerability in daily life: an experience sampling test study. AB - BACKGROUND: Epidemiological findings suggest that cannabis use is a risk factor for the emergence of psychosis, and that the induction of psychotic symptoms in the context of cannabis use may be associated with a pre-existing vulnerability for psychosis. This study investigated in a non-clinical population the interaction between cannabis use and psychosis vulnerability in their effects on psychotic experiences in daily life. METHOD: Subjects (N = 79) with high or low levels of cannabis use were selected among a sample of 685 undergraduate university students. Experience sampling method (ESM) was used to collect information on substance use and psychotic experiences in daily life. Vulnerability to develop psychosis was measured using a clinical interview assessing the level of psychotic symptoms. Statistical analyses were performed using multilevel linear random regression models. RESULTS: The acute effects of cannabis are modified by the subject's level of vulnerability for psychosis. Subjects with high vulnerability for psychosis are more likely to report unusual perceptions as well as feelings of thought influence than subjects with low vulnerability for psychosis, and they are less likely to experience enhanced feelings of pleasure associated with cannabis. There is no evidence that use of cannabis is increased following occurrence of psychotic experiences as would be expected by the self-medication model. CONCLUSION: Cannabis use interacts with psychosis vulnerability in their effects on experience of psychosis in daily life. The public health impact of the widespread use of cannabis may be considerable. PMID- 12537034 TI - Double-blind controlled investigation of bilateral prefrontal transcranial magnetic stimulation for the treatment of resistant major depression. AB - BACKGROUND: The efficacy and safety of bilateral prefrontal repetitive transcranial magnetic stimulation (rTMS) for treating resistant major depression were examined in a double-blind, placebo-controlled study. METHOD: Nineteen medication-resistant depressed subjects were randomly assigned to 3 weeks of active or sham rTMS. Effects on mood and neuropsychological function were assessed. RESULTS: Both groups improved significantly in mood over the 3 weeks, but there was no significant difference between active and sham treatments. There were no significant neuropsychological effects. CONCLUSIONS: Bilateral rTMS was not superior to sham in treating resistant depression in this pilot study, but caused no neuropsychological impairment. PMID- 12537035 TI - Effects of acute tryptophan depletion on cognitive function in Alzheimer's disease and in the healthy elderly. AB - BACKGROUND: The cholinergic system is profoundly impaired in senile dementia of Alzheimer type (SDAT) and replacement therapy produces only modest clinical benefits. The serotonergic system is also impaired and may contribute both to cognitive and non-cognitive symptoms in SDAT. To investigate this further we assessed the effects of lowering brain serotonin using the technique of acute tryptophan depletion on cognitive function in patients with SDAT and in age matched control subjects. METHOD: Sixteen patients with probable SDAT and 17 healthy elderly subjects received two amino acid drinks in a within subject, double-blind, placebo-controlled, counterbalanced, crossover design. One of the drinks was nutritionally balanced and contained tryptophan (placebo), the other was identical but contained no tryptophan. A battery of detailed neuropsychological tests was performed between 4 and 6 h after the drink. Mood rating scales and other ratings of behavioural and emotional symptoms were also performed on both occasions. RESULTS: Acute tryptophan depletion resulted in impairment on tasks of working memory in both groups. There was no group specific effect. Female SDAT subjects performed better on a task of pattern recognition during acute tryptophan depletion compared with placebo. There were no changes in behavioural symptoms during acute tryptophan depletion in either group. CONCLUSION: Compromised serotonergic function may be an important contributor to cognitive decline in SDAT and in ageing. Strategies targeting specific 5HT receptors may be helpful in SDAT. PMID- 12537036 TI - Symptom fluctuation in bulimia nervosa: relation to menstrual-cycle phase and cortisol levels. AB - BACKGROUND: Individuals with bulimia nervosa report significant symptom fluctuation, and some studies have suggested a premenstrual exacerbation of binge frequency. The purpose of this study is to explore the hormonal correlates of symptom fluctuation in bulimia nervosa. METHOD: For five consecutive weeks (one full menstrual cycle), eight women with bulimia nervosa and eight non-eating disordered control women collected morning saliva samples and recorded several mood characteristics; the bulimic women also recorded binge and purge episodes. Subsequently, salivary cortisol and androgen levels were determined by radioimmunoassay. RESULTS: Bulimic symptoms were exacerbated in both the mid luteal and premenstrual phases, when compared with the follicular and ovulatory phases (F(3,21) = 3.76, P = 0.026; contrast analysis t(7) = 3.47, P < 0.01). Fluctuation in cortisol was closely correlated with fluctuation of bulimic symptoms, with elevatedcortisol secretion following symptom exacerbation (r(24) = 0.64, P = 0.001). CONCLUSIONS: Bulimic symptom fluctuation appears to be related to two hormonal phenomena--phase of the menstrual cycle and cortisol secretion- with menstrual-cycle phase influencing bulimic symptom severity, and bulimic symptom severity effecting increases in cortisol secretion. Improved understanding of the hormonal causes and consequences of symptom fluctuation may lead to improved psychological and pharmacological treatments for bulimia nervosa. PMID- 12537037 TI - Vulnerability and resiliency to suicidal behaviours in young people. AB - BACKGROUND: We aimed to examine factors that influence vulnerability/resiliency of depressed young people to suicidal ideation and suicide attempt. METHOD: Data were gathered during a 21-year longitudinal study of a birth cohort of 1,265 New Zealand young people. Measures included: suicide attempt; suicidal ideation; major depression; childhood, family, individual and peer factors. RESULTS: Young people who developed major depression had increased rates of suicidal ideation (OR = 54: 95% CI 4.5-6.6) and suicide attempt (OR = 12.1; 95% CI 7.9-18.5). However, the majority of depressed young people did not develop suicidal ideation or make suicide attempts, suggesting that additional factors influence vulnerability or resiliency to suicidal responses. Factors influencing resiliency/vulnerability to suicidal responses included: family history of suicide; childhood sexual abuse; neuroticism; novelty seeking; self-esteem; peer affiliations; and school achievement. These factors operated in the same way to influence vulnerability/resiliency among those depressed and those not depressed. CONCLUSIONS: Vulnerability/resiliency to suicidal responses among those depressed (and those not depressed) is influenced by an accumulation of factors including: family history of suicide, childhood sexual abuse, personality factors, peer affiliations and school success. Positive configurations of these factors confer increased resiliency, whereas negative configurations increase vulnerability. PMID- 12537038 TI - Factors associated with the wish to hasten death: a study of patients with terminal illness. AB - BACKGROUND: There is considerable debate regarding the clinical issues surrounding the wish to hasten death (WTHD) in the terminally ill. The clinical factors contributing to the WTHD need further investigation among the terminally ill in order to enhance understanding of the clinical assessment and treatment needs that underlie this problem. A more detailed understanding may assist with the development of appropriate therapeutic interventions. METHOD: A sample of terminally ill cancer patients (N = 256) recruited from an in-patient hospice unit, home palliative care service and a general hospital palliative care consulting service from Brisbane Australia between 1998-2001 completed a questionnaire assessing psychological (depression and anxiety), social (family relationship, social support, level of burden on others) and the impact of physical symptoms. The association between these factors and the WTHD was investigated. RESULTS: A high WTHD was reported by 14 % of patients. A discriminant function analysis revealed that the following variables were associated with a high WTHD (P < 0.001): higher levels of depressive symptoms, being admitted to an in-patient hospice setting, a greater perception of being a burden on others, lower family cohesion, lower levels of social support, higher levels of anxiety and greater impact of physical symptoms. CONCLUSIONS: Psychological and social factors are related to a WTHD among terminally ill cancer patients. Greater attention needs to be paid to the assessment of psychological and social issues in order to provide appropriate therapeutic interventions for terminally ill patients. PMID- 12537039 TI - Psychiatrists' attitudes to maintenance medication for patients with schizophrenia. AB - BACKGROUND: Maintenance antipsychotic medication is the mainstay of relapse prevention in patients with schizophrenia. Long acting depot antipsychotics were developed to promote treatment adherence and yet their utilization is variable, perhaps due to negative attitudes of both patients and psychiatrists. Recently, a shift away from depots has occurred, in favour of the newer atypical oral antipsychotics. METHOD: This study investigated the current attitudes and knowledge concerning depots, with a newly designed questionnaire, in a cross sectional postal survey of qualified psychiatrists working in south-east England. RESULTS: A substantial minority of psychiatrists believe that depots are old fashioned (40%), stigmatizing (48%) and are associated with more side-effects than typical oral antipsychotics (38%). Many believe that depots are as efficacious as oral medication (91%) but are less acceptable to patients (69%) and relatives (66%). A large majority consider depots enhance patient compliance (81%) and prevent relapse (94%). Psychiatrists would be persuaded to prescribe depots if they were associated with fewer side-effects, in patients where compliance is an issue, and if atypical depot antipsychotics were available, presumably because they would have a lower incidence of side-effects. Additionally, psychiatrists' knowledge about depots was positively associated with attitudes. More favourable patient-centred attitudes were reported by psychiatrists with higher depot use. CONCLUSION: Practising psychiatrists have several strongly endorsed attitudes towards depot medication that are associated with knowledge and prescribing habits. By updating psychiatrists' knowledge about depots, in turn their attitudes may become more positive and prescribing practices may subsequently change. PMID- 12537040 TI - The family and compliance in schizophrenia: the influence of clinical variables, relatives' knowledge and expressed emotion. AB - BACKGROUND: . A variety of factors are related to compliance with medication in schizophrenia, but little attention has been paid to the role of families. Carers' knowledge or expressed emotion (EE) may be related to compliance. The aim of the present study was to evaluate the relevance of these two factors, as well as their relationships with other variables for the prediction of compliance. METHOD: A sample of patient-carer pairs (N = 79) involved in a family intervention for schizophrenia trial was recruited. Compliance, symptoms, social functioning and attitudes to their carers were assessed in patients. Carers' EE, knowledge and psychopathology were also evaluated. RESULTS: A number of factors were related to compliance, including carers' EE and patients' psychotic symptoms, which contributed independently to not taking medication. Carers' knowledge about schizophrenia and other groups of symptoms was not related to compliance. CONCLUSIONS: EE may be an important factor to account for in the understanding of patients' compliance and the direction of the relationship between EE and compliance should be the subject of further study. PMID- 12537041 TI - Does treatment delay in first-episode psychosis really matter? AB - BACKGROUND: Relatively few predictors of outcome in first-episode psychosis are potentially malleable and duration of untreated psychosis (DUP) is one. However, the degree to which DUP is mediated by other predictors of outcome is unclear. This study examines the specific effects of DUP on 12-month outcome after adjusting for effects of potential confounders and moderating variables. METHOD: The sample comprised 354 first-episode psychosis patients followed up 12-months after remission/stabilization of their psychotic symptoms. Outcome measures included functional outcome, severity of positive symptoms and negative symptoms. Hierarchical multiple regression assessed whether DUP significantly predicted 12 month outcome after adjusting for other predictors. Contrast analysis further clarified the differential effects of DUP on 12-month outcome. RESULTS: DUP remained a significant predictor of outcome after adjusting for the effects of other variables. This finding remained robust for the subset of patients with schizophrenia or schizophreniform disorder. Functional outcome appeared to decline substantially even after very short treatment delays (> 7 days), with more gradual deterioration in functioning until very long DUP (> 1 year). Good outcome was variably associated with good pre-morbid adjustment, female gender, diagnosis of affective disorder, short duration of prodromal symptoms, and treatment within the Early Psychosis Prevention and Intervention model in contrast to other models of care. CONCLUSIONS: DUP consistently predicts outcome independently of other variables, and is not simply a proxy for other factors. As one of the few potentially malleable factors influencing outcome, DUP could prove to be a target for secondary preventive efforts in early psychosis. PMID- 12537042 TI - Insight: its relationship with cognitive function, brain volume and symptoms in schizophrenia. AB - BACKGROUND: Lack of insight is frequently observed in schizophrenia. Relationships have been noted between poor insight, clinical symptoms and cognitive impairments but the findings are inconsistent. There have been some recent attempts to relate poor insight to neuro-anatomical measures. METHOD: We assessed insight, positive and negative symptoms of schizophrenia, cognitive performance, and whole brain volumes in a sample of 78 DSM-IV male schizophrenics and 36 normal male comparison subjects matched for age and IQ. Subjects underwent a dual-echo MRI brain scan to establish grey, white and whole brain volumes. RESULTS: Poor Wisconsin Card Sorting Test performance inversely correlated with insight in schizophrenic patients, as did the symptoms alogia, anhedonia, avolition/apathy, affective flattening, inappropriate affect, thought disorder and delusions. The presence of inappropriate affect, delusions and thought disorder, showed the most significant impact on insight levels. There were no significant correlations between whole brain, white and grey matter volume and degree of insight. CONCLUSIONS: The results suggest that poor insight is significantly related to schizophrenic psychopathology, and confirm that there is a relationship between insight and executive performance. Awareness of illness is not related to any global brain measures, suggesting future investigations should pay attention to more specific cortical regions such as the frontal cortex. PMID- 12537043 TI - Impaired distractor inhibition in patients with schizophrenia on a negative priming task. AB - BACKGROUND: Numerous studies have suggested, via the interpretation of negative priming effects, that subjects with schizophrenia are less able than controls to inhibit irrelevant distracting information. Further issues concerning impairment in inhibitory processes are investigated here. First, recent research has revealed that negative priming (NP) effects can be caused by different processes, distractor inhibition or perceptual review. Therefore, conclusions concerning reduced inhibition in patients with schizophrenia are not possible from previous NP research. Secondly, previous NP studies have required subjects to identify some feature of the target. This is the first study to examine NP that uses a spatial task in patients with schizophrenia. METHOD: Twenty-eight subjects with schizophrenia and 28 age and sex matched non-psychiatric control subjects completed a computerized NP task that eliminated the possible contribution of perceptual review. RESULTS: Subjects with schizophrenia had reduced levels of NP compared to control subjects on this spatial NP task (t = 2.46, P < 0.02). Current age, positive, negative or total PANNS scores did not correlate with negative priming scores, but post hoc analyses revealed that clozapine-treated patients had significantly greater levels of negative priming than patients receiving typical antipsychotic medications. CONCLUSIONS: The present experiment eliminated the contribution of perceptual review to negative priming and demonstrated that when a pure measure of inhibition is taken on a localization task, patients with schizophrenia were less able to inhibit irrelevant distracting stimuli. The fact that NP was reduced in a spatial task suggested a more diffuse reduction in inhibition than previous studies that examined only identification-based responses. PMID- 12537044 TI - Source monitoring and memory confidence in schizophrenia. AB - BACKGROUND: The present study attempted to extend previous research on source monitoring deficits in schizophrenia. We hypothesized that patients would show a bias to attribute self-generated words to an external source. Furthermore, it was expected that schizophrenic patients would be overconfident regarding false memory attributions. METHOD: Thirty schizophrenic and 21 healthy participants were instructed to provide a semantic association for 20 words. Subsequently, a list was read containing experimenter- and self-generated words as well as new words. The subject was required to identify each item as old/new, name the source. and state the degree of confidence for the source attribution. RESULTS: Schizophrenic patients displayed a significantly increased number of source attribution errors and were significantly more confident than controls that a false source attribution response was true. The latter bias was ameliorated by higher doses of neuroleptics. CONCLUSIONS: It is inferred that a core cognitive deficit underlying schizophrenia is a failure to distinguish false from true mnestic contents. PMID- 12537045 TI - Behavioural phenotypes associated with specific genetic disorders: evidence from a population-based study of people with Prader-Willi syndrome. AB - BACKGROUND: Prader-Willi syndrome (PWS) is a genetic disorder resulting in obesity, short stature, cryptorchidism, learning disabilities (mental retardation) and severe neonatal hypotonia. Associated with the syndrome are a number of behaviours that are sufficiently distinctive that the syndrome is considered to have a specific 'behavioural phenotype'. METHODS: Through multiple sources we attempted to identify all people with PWS living in one region in the U K. This cohort was augmented by people with PWS from other regions, and a contrast group of people with learning disabilities of varied aetiologies. The main carers were interviewed, using structured and semi-structured interview schedules, to establish the presence and severity of specific behaviours, and PWS diagnostic criteria. The intellectual functioning and attainments of all were determined. Blood samples were obtained for genetic diagnosis from all consenting participants. RESULTS: Although excessive eating was recognized as a potentially severe problem in those with PWS, it was almost universally controlled by food restriction, and therefore not seen as a 'problem behaviour'. Those with PWS differed from a learning disabled group of other aetiologies in the prevalence rates of skin picking, temper tantrums, compulsive behaviours and mood fluctuations, and also in the profile of their adaptive behaviours. CONCLUSIONS: The study confirms the distinct behavioural phenotype of PWS. Specific behaviours occurred significantly more frequently in PWS, compared with an age and BMI matched learning disabled comparison group. A factor analysis of the behaviours involved resulted in three factors that we hypothesized to be independent, and to arise from different mechanisms. PMID- 12537046 TI - The health effects of peace-keeping in the UK Armed Forces: Bosnia 1992-1996. Predictors of psychological symptoms. AB - BACKGROUND: Peace-keeping duties are associated with unique stresses for military personnel. There have been few reports on the effects of peace-keeping on psychological health. METHOD: We used data from a cross-sectional study originally established to examine health effects of service in the Gulf War, which included a control group who had been deployed in Bosnia (N= 2049). This group was examined to establish which demographic, occupational, and deployment related risk factors were associated with psychological distress measured on the General Health Questionnaire (GHQ-12) and stress symptoms akin to a post traumatic stress reactions. RESULTS: The main risk factors for stress symptoms in the Bosnia group were--lower rank, being deployed early in the campaign, having more deployment-related exposures, and serving on staff duties. There was no protective effect of previous deployments to the Falklands or Northern Ireland, and time off following deployment was not protective. For GHQ-12 cases, the main risk factors were: female gender, lower rank, increased deployment-related exposures and serving on staff duties. CONCLUSIONS: This study suggests that stress symptoms and GHQ-12 cases, are strongly predicted both by experiences during deployment and demographic factors. PMID- 12537047 TI - Seasonality of symptom onset in first-episode schizophrenia. AB - BACKGROUND: There have been numerous reports of seasonal trends in psychotic illnesses. In schizophrenia, seasonal trends in incidence have been shown to be especially apparent in first-episode cases. Most previous research has used date of admission as a proxy for date of incidence of disorder; we present results of an investigation into seasonal trends in dates of onset of symptoms in a group of 295 first-episode cases of schizophrenia and schizophreniform disorder. METHOD: Data were analysed using statistical methods appropriate for detecting seasonal trends in pooled data over 6 years. RESULTS: Only male cases of schizophrenia and schizophreniform disorder showed a significant seasonal distribution to dates of onset of symptoms, with a peak in August (winter). CONCLUSION: True seasonal patterns are present in schizophrenia incidence, but their consistency with other published studies and wider significance, is difficult to ascertain because of different methods used in dating incidence of disorder. PMID- 12537048 TI - Implicit memory for words presented in short texts is preserved in Alzheimer's disease. AB - BACKGROUND: The level of efficiency of implicit memory in Alzheimer's disease remains unclear as previous studies using stem completion tasks have led to contradictory results. METHOD: The present study used target words embedded in significant short texts that subjects were required to read aloud (i.e. to enhance semantic processing). Texts were presented in two perceptual situations: 'simple' (blank spaces delimitating words) and complex' (spaces were filled by '8's). In the completion phase, patients had to write the first word that came to mind in order to complete a three-letter stem. The recognition phase explored explicit memory performance. The performance of 24 Alzheimer patients was compared to a matched sample of healthy controls. RESULTS: Reading times differed between groups and were shorter for healthy controls. Recognition was dramatically lower in patients, thus confirming the alteration of explicit memory in this pathology. However, a significant priming effect (e.g. the tendency to complete the stem with the aid of a previously explored word) was present in both groups and did not differ between patients and healthy controls. CONCLUSIONS: The absence of a correlation between priming and recognition scores suggests that this result cannot be explained by an explicit memory bias. Moreover, as the priming level was identical whatever the perceptual aspect of the text, we suggest that the priming effect is not only mediated by perceptual processes but also by lexical and conceptual processes, which to some extent are preserved during the light and moderate stages of this disease. PMID- 12537049 TI - Surgery for pediatric vocal cord paralysis: a retrospective review. AB - To determine the outcome of surgical procedures for bilateral vocal cord paralysis in children, we performed a retrospective review of children under 18 years of age with bilateral vocal cord paralysis and a previous tracheotomy who underwent a primary procedure at a single tertiary care institution with an aim of decannulation. The primary outcome measure was the operation-specific decannulation rate (OSDR). The overall decannulation rates, as well as morbidity rates, were also recorded. Fifty-two children met the inclusion criteria (mean age at time of primary surgery, 6.2 years; SD, 5 years). Vocal cord lateralization procedures combined with a partial arytenoidectomy achieved the highest OSDR (17/24 or 71%). This OSDR was statistically higher than the OSDRs for CO2 laser cordotomy or arytenoidectomy procedures (OSDR, 5/17 or 29%, p = .008), for isolated arytenoidopexy procedures (OSDR, 1/4 or 25%, p = .000004), or for posterior costal cartilage graft procedures (OSDR, 3/5 or 60%, p = .0004). Neither of the 2 children who underwent isolated arytenoidectomy achieved primary decannulation. The incidence of aspiration following posterior cartilage graft procedures was 15% (2/15). Subanalysis by age failed to reveal differences in OSDR. We conclude that vocal cord lateralization procedures with partial arytenoidectomy afford the highest OSDR among primary procedures for pediatric vocal cord paralysis. The CO2 laser procedures, while having limited success as a primary procedure, are effective for revision. PMID- 12537050 TI - Treatment of recurrent respiratory papillomatosis in children with the microdebrider. AB - The mainstay of treatment of children with recurrent respiratory papillomatosis has been CO2 laser laryngoscopy. Powered microdebriders allow precise removal of laryngeal lesions without thermal damage. The microdebrider may reduce costs by reducing operative time and limiting the need for expensive laser-related equipment. To compare the use of the microdebrider with that of the CO2 laser in children undergoing surgery for recurrent respiratory papillomatosis, we retrospectively reviewed the charts of 18 patients from 2 institutions covering a treatment period between December 1998 and October 2001. These patients were initially treated with the CO2 laser, but were switched to microdebrider resection during the review period. We recorded the number of procedures, the technique(s) of resection used, and the operative time for each procedure. We identified 18 patients, 11 male and 7 female, ages 18 months to 14 years. A total of 127 procedures were performed with the CO2 laser, and a total of 50 procedures were performed with the microdebrider. The mean number of procedures per patient was 10 (range, 2 to 17 procedures). The mean surgical time per procedure using the CO2 laser was 59.2 minutes (range, 8 to 130 minutes). The mean time per procedure with the microdebrider was 32.4 minutes (range, 12 to 47 minutes). This reduction in operative time was clinically and statistically significant (p = .0001, unpaired Student's t-test). We concluded that microdebrider resection of laryngeal papilloma in children allowed more rapid surgery with potentially reduced treatment costs. Other advantages of this technique include precise excision without thermal injury. Our report details useful refinements of technique with the microdebrider. PMID- 12537051 TI - Use of the endoscopic microdebrider in the management of airway obstruction from laryngotracheal carcinoma. AB - Endoscopic debridement of obstructing laryngotracheal carcinoma with the microdebrider is an alternative to CO2 laser excision or emergency tracheotomy. This is a retrospective study of 27 patients with advanced laryngotracheal carcinoma treated with the microdebrider for airway obstruction from 1998 to 2002. In 26 patients, the carcinoma was debrided successfully and tracheotomy was safely avoided. Postoperative bleeding was encountered in 1 patient with a coagulopathy. Two of the patients later required tracheotomy during radiotherapy secondary to radiation-induced laryngeal edema. Both were safely managed without acute airway compromise. Laryngeal debridement provided up to 3 months of stable airway until definitive oncological treatment was performed. Endoscopic microdebridement of obstructing carcinoma is a relatively simple and highly accurate approach to restoring a stable airway. It provides extra time for preoperative oncological, psychological, and nutritional planning. PMID- 12537052 TI - Topical nasal anesthesia and laryngopharyngeal sensory testing: a prospective, double-blind crossover study. AB - Laryngopharyngeal sensory discrimination testing (LPSDT) has become a popular means of detecting sensory deficits in patients with voice and/or swallowing complaints. During this procedure, transnasal fiberoptic laryngoscopy (TFL) using a specialized fiberoptic scope is performed in order to deliver discrete pulses of air to the laryngopharyngeal mucosa. Historically, topical anesthesia has been applied to the nasal mucosa to lessen discomfort during TFL. Because of the concern that topical nasal anesthesia could also anesthetize the laryngopharynx, it is usually not used during sensory testing. To prospectively compare LPSDT scores after the nasal administration of cocaine hydrochloride, oxymetazoline hydrochloride, and saline solution, we prospectively evaluated 15 subjects undergoing LPSDT. Each had the procedure performed on 3 separate occasions. Before examination, each patient was administered either 4% cocaine, 0.05% oxymetazoline, or saline solution by atomizer (2 sprays of 1-second duration to each naris). By the conclusion of the study, each patient had undergone sensory testing with each of the test agents. During each examination, the minimal air pulse eliciting the laryngeal adductor reflex was recorded for both sides of the laryngopharynx. Both patient and examiner were blinded to the test agent used. The mean sensory score for saline solution was 3.9 mm Hg. The sensory scores for oxymetazoline and cocaine were 3.4 and 3.5 mm Hg, respectively. When compared to saline solution and oxymetazoline independently, sensory testing of the laryngopharynx during which the nose was anesthetized with cocaine resulted in similar thresholds for eliciting the laryngeal adductor reflex (p = .40 and p = .85). We conclude that topical anesthesia can be applied to the nasal cavity during LPSDT without altering laryngeal sensation. PMID- 12537053 TI - Simple technique for tracheocutaneous fistula closure in the pediatric population. AB - To review our experience with cauterization of persistent tracheocutaneous fistulas in children, we performed a retrospective review of patients who underwent cauterization of tracheocutaneous fistulas by the senior author (O.E.B.) from 1986 to 2001 in an academic, tertiary care children's hospital. We studied 13 pediatric patients ranging in age from 2.5 to 17.5 years of age at the time of surgery. Twelve patients underwent cauterization under endoscopic visualization. One patient underwent superficial cauterization of the tract without endoscopy. All patients had at least a 1-year history of an indwelling tracheotomy. All patients were decannulated at least 1 year before fistula cauterization. Of the 12 patients who underwent intraoperative airway endoscopy, the internal orifice of the fistula tract was specifically visualized and seen to be patent in 10. One patient was noted to have internal mucosalization of the tract, and no discrete opening to the trachea was noted in the other patient. Eleven patients had complete closure of the fistula site at follow-up (range, 2 weeks to 2 years). One patient developed a leak during a coughing spell 2 days after the operation, and the fistula was noted to be closing with a small leak at follow-up. Another patient (with Treacher Collins syndrome) ultimately required a repeat tracheotomy for persistent obstructive sleep apnea. This patient was the only one admitted after the operation, for a pulmonary infiltrate. No other patients required airway support in the immediate postoperative period. Cauterization of tracheocutaneous fistulas in children is a relatively simple, effective, and safe technique. PMID- 12537054 TI - Postoperative dysphagia versus neurogenic dysphagia: scintigraphic assessment. AB - In order to differentiate the features of dysphagia that occur after supraglottic horizontal laryngectomy from those that occur during neurologic diseases, we divided 38 subjects into 3 groups and submitted them to oropharyngoesophageal scintigraphy. Group 1 (control group) included 15 healthy volunteeers; group 2 comprised 8 patients who had residual dysphagia at least 1 year after supraglottic laryngectomy; and group 3 included 15 patients with various neurologic and neuromuscular disorders. In group 1, the mean values (+/- 2 SD) of selected semiquantitative parameters were consistent with those reported in the literature for normal subjects. In group 2, oral, pharyngeal, and esophageal transit times were not significantly altered, and moderate tracheobronchial post deglutitive aspiration was present (maximum value, 6.7%; mean value, 2.04%). The pharyngeal retention index was significantly increased (p = .0003) as compared to normal subjects in all cases (maximum value, 40%; mean value, 23%) and was associated in all cases with slight but consistent post-deglutitive aspiration. In group 3, the oral and esophageal phases were significantly prolonged and the retention indices were significantly increased. Statistical analysis documented a significant increase in oral transit time (p = .003), esophageal transit time (p = .01), oral retention index (p = .006), pharyngeal retention index (p = .0007), and esophageal retention index (p = .009) as compared to normal subjects. The swallowing pattern was also altered by 1) an early loss of the bolus from the oral cavity; 2) bolus fragmentation due to double or triple deglutition, reduced lingual propulsion, or the return of a small part of the bolus into the oral cavity during deglutition; and/or 3) double pharyngeal peaks in the activity-time curves. Tracheobronchial aspiration (maximum value, 90%; mean value, 9.70%) was present in some cases, mainly in patients affected by post-stroke dysphagia. On the basis of the obtained results and considering the low doses of radiation delivered to the patient (0.043 Gy), the limited invasiveness, and the excellent patient tolerance, scintigraphy appears to be clinically valid in the functional study of swallowing and in identifying different deglutition disorders. PMID- 12537055 TI - Endoscopic treatment of upper airway and digestive tract lesions caused by caustic agents. AB - We present our experience in the endoscopic management of upper aerodigestive tract lesions caused by caustic agents. Between 1985 and 2000, 112 patients with upper airway and digestive tract lesions due to caustic agents were treated at the Airways Endoscopic Surgery Unit of Padua Hospital: 44 were male and 68 were female, and the median age was 42.6 years. A retrospective statistical analysis of our case series was made to evaluate the factors that most influenced the severity of injuries and the outcome of treatment. All of the patients underwent videoendoscopic assessment. In 79 cases, the patients came under our observation in the acute phase, whereas 33 presented a picture of chronic lesions. Acute lesions were classified into 3 grades. All acute grade 1 lesions healed spontaneously. In the 32 patients with grade 2 lesions, endoscopic treatment based on removal of necrotic tissue, dilations, and cleansing of abnormal fibrin adhesions resolved the disease in 30 cases (94%). Nine patients (43%) with acute grade 3 lesions developed severe chronic lesions that required subsequent treatments. Rigid endoscopy with diode laser-assisted radial lysis was performed in 32 patients with chronic cicatricial lesions and was successful in 30 cases (94%). We reiterate the need for a standardized multidisciplinary protocol for treating lesions caused by caustic agents and emphasize the essential role of airway and digestive canal videoendoscopy in the diagnosis and treatment of both the acute lesions and chronic cicatricial sequelae. PMID- 12537056 TI - New therapy for orolaryngeal manifestations of Crohn's disease. AB - Crohn's disease is a chronic inflammation that may involve the entire gastrointestinal tract, from the mouth to the anus. The most widely accepted etiologic theory involves an immunologic aberration leading to local tissue destruction. Cell-mediated immunity with increased tumor necrosis factor (TNF) production may play a role in mucosal damage. Oral and laryngeal involvement are rare manifestations of Crohn's disease that are usually treated successfully by steroids. We here report a rare case of extra-intestinal Crohn's disease resistant to steroid therapy, which was successfully treated with infliximab, a chimeric antibody directed against TNF-alpha that is the only registered agent for the treatment of Crohn's disease. The relative safety, efficacy, and efficiency of infliximab make it an alternative treatment of which otolaryngologists should be aware. PMID- 12537057 TI - Predictive value of sleep nasendoscopy in the management of habitual snorers. AB - Sleep nasendoscopy (SN) is claimed to identify the site of obstruction in habitual snoring and is used to select patients for palatal surgery. The validity of SN is, however, unproven. This is the first prospective blind assessment of the role of SN in the management of habitual snorers. Fifty-five consecutive patients listed for laser-assisted uvulopalatoplasty (LAUP) for simple snoring on clinical criteria underwent SN immediately before surgery. Snoring severity and Epworth Sleepiness Scales were completed before the operation and at least 3 months after the operation. The postoperative scores on the snoring severity scale had significantly improved after LAUP, for each SN grade: grade 1 (palatal obstruction; p = .04) and grade 2 (multisegmental obstruction; grade 2A, p = .003, and grade 2B, p = .01). The Epworth Sleepiness Scale showed significant improvement in the whole group (p = .02). We conclude that SN grading of sites of pharyngeal collapse is an insufficient basis to exclude patients from LAUP. Sleep nasendoscopy is thus not of value in the management of habitual snoring, as the measure is a poor predictor of favorable outcome after palatal surgery. PMID- 12537058 TI - Passive marker computer-aided sinonasal and cranial base surgery: observations from a learning curve. AB - To assess the feasibility of passive marker computer-aided surgery in a single institution, we performed 22 procedures in 21 patients with disorders including sinonasal tumors (n = 9), fungal sinusitis (n = 4), recurrent polyps (n = 3), chronic sinusitis (n = 3), and cerebrospinal fluid rhinorrhea (n = 2). Passive marker computer-aided surgery was successful in 19 of the 21 patients. The accuracy was on the order of 1.35 mm. Probe conversion, rotation, and cordlessness were helpful in all 19 cases. The system helped with landmarks (n = 14), margins (n = 7), skull base (n = 6), orbit (n = 5), and approach (n = 4). Computer-aided surgery accurately confirmed the location of an instrument and demonstrated tumor-normal tissue interfaces. It aided the surgeon in procedures on the sinonasal area and cranial base. The potential advantages of a passive marker system as compared with other available technologies center around the ability to convert and/or rotate virtually any instrument to a cordless imaging probe on demand during the operation. PMID- 12537059 TI - Rate of recovery of taste function after preservation of chorda tympani nerve in middle ear surgery with special reference to type of disease. AB - To study the recovery of function of the chorda tympani nerve, we examined by electrogustometry 79 patients (83 ears) with both preoperative normal function of the nerve and operative preservation of the nerve, every 2 days during hospitalization and at 6 months after surgery. For symptoms such as tongue numbness and taste disturbance, patients with noninflammmatory (NI) diseases (13/20 or 65.0%) showed a significantly higher rate of symptoms than did patients with chronic otitis media (COM; 13/35 or 37.1%) at 2 weeks after surgery (p = .032). The patients with NI diseases (5/20 or 25.0%) tended to show a higher rate of symptoms than did the COM patients (2/35 or 5.7%) or cholesteatoma patients (2/28 or 7.1%) at 6 months after surgery. The rate of recovery of the EGM threshold to normal at 2 weeks after surgery was significantly lower in NI disease patients (6/20 or 30.0%) than in COM patients (23/ 35 or 62.9%) or cholesteatoma patients (19/28 or 67.9%; p = .015 and .008, respectively). Thus, the patients with NI diseases had postoperative symptoms and elevation of EGM threshold more frequently than did the patients with inflammatory diseases. PMID- 12537060 TI - Minimally invasive orbital decompression for Graves' ophthalmopathy. AB - We reviewed a 7-year experience at a tertiary-care, academic medical center with balanced, minimally invasive decompression for Graves' ophthalmopathy, in an effort to define the goals, risks, and outcomes of surgical intervention. Endoscopic medial decompression was performed in 26 patients; 23 underwent lateral decompression as well, and 13 also had inferior decompression. Septoplasty, turbinate reduction, and orbital rim augmentation were performed as needed. The indications for surgery were threat to vision (n = 10) and proptosis with a desire to return to the predisease state (rehabilitative, n = 16). The exophthalmos improved by a mean of 4.4 mm (p < .001). All patients who had surgery for threatened vision had improved vision after the operation. There were 3 patients with new-onset postoperative diplopia, 2 of whom underwent strabismus surgery. There was 1 case of postoperative sinusitis, which resolved with oral antibiotics and nasal decongestion, and 1 case of transient ulnar neuropathy. There were no other intraoperative or postoperative complications. Modern methods of orbital decompression provide a minimally invasive, effective, and relatively safe approach to the treatment of Graves' ophthalmopathy. PMID- 12537061 TI - Post-tonsillectomy and -adenoidectomy hemorrhage in nonselected patients. AB - This retrospective study was undertaken to evaluate the incidence of hemorrhage in nonselected patients requiring surgical treatment after adenoidectomy (group 1; 7,785 patients) or tonsillectomy with or without adenoidectomy (group 2; 6,794 patients). Postoperative bleeding from the tonsillar fossae occurred in 200 patients (2.94%). Primary hemorrhage (< 24 hours) clearly prevailed in groups 1 (86%) and 2 (78%). There was 1 case with a lethal outcome following tonsillectomy (0.007%). Bleeding from the epipharynx occurred in 17 patients of group 1 (0.21%). A total of 5 patients received blood transfusions, all of them belonging to group 2 (0.07%). The latest bleeding occurred 6 days (adenoidectomy) and 18 days (tonsillectomy) after surgery. Hemorrhage following both procedures is a rare complication predominantly occurring several hours after surgery and in nonselected patients. Male gender, age over 70 years, infectious mononucleosis, and a history of recurrent tonsillitis were found to be risk factors for posttonsillectomy hemorrhage. PMID- 12537062 TI - Tongue representation in the lateral cortical motor region of the human brain as assessed by transcranial magnetic stimulation. AB - Motor evoked potentials (MEPs) of the lingual muscles elicited by focal cortical transcranial magnetic stimulation (TMS) with a figure 8-shaped coil were investigated in 17 healthy subjects with special regard to amplitude and onset latency as a function of the coil position on the head surface. Bilateral reproducible responses could be observed at coil positions mostly varying from approximately 6 to 13 cm lateral to the vertex. During moderate muscle activation, maximum responses were obtained at a mean stimulus position of about 11 cm lateral and 3 cm anterior to the vertex with similar onset latencies, but with significantly higher amplitudes on the contralateral side (3.5 +/- 1.9 mV, 9.5 +/- 1.1 ms contralateral; 2.6 +/- 1.5 mV, 9.7 +/-1.0 ms ipsilateral). Comparing our data on the orbicularis oculi muscle with the results obtained on lower lip muscles, we found a considerable overlap of those coil positions from which reproducible MEPs could be elicited in both groups of target muscles, but the lingual area was placed more laterally. Thus, a statistically significant separation of the cortical representation areas of lingual and lower lip mimetic muscles is possible by focal cortical TMS, reflecting somatotopic organization of the face-associated motor cortex. PMID- 12537063 TI - Nicotinic cholinergic receptor expression in the human nasal mucosa. AB - Twenty-four nasal mucosa specimens were obtained from the inferior or middle turbinates of 6 normal subjects and 18 patients with chronic sinusitis, inflammatory polyp formation, or sinus allergies. Reverse transcription polymerase chain reaction analysis was used to identify the non-neuronal nicotinic cholinergic receptor (nAChR) subunits that were expressed in the nasal mucosa. Collectively, transcripts for alpha (alpha1, alpha2, alpha3, alpha4, alpha6, alpha7) and beta (beta2, beta3, beta4) nAChR subunit genes were detected in the respiratory mucosa. The alpha3, alpha7, and beta2 subunits were expressed in 92%, 88%, and 75% of the subjects, respectively. There was a high degree of interindividual variation in nAChR subunit gene expression among subjects. A significant univariate association was found between tissue type and beta4 expression and between gender and beta3 expression. These data suggest that cells in the nasal mucosa express the necessary messenger RNAs (mRNAs) for numerous nAChR combinations. Moreover, our identification of nAChR subunit mRNAs in the nasal mucosa extends the findings of other functional studies of nAChRs in nasal epithelial cells and implies that nicotine from tobacco products such as cigarette smoke and nicotine nasal spray may have direct cellular effects on nasal mucosa cells through activation of homogeneous or heterogeneous nAChRs. A significant number of patients receiving nicotine nasal spray have reported nasal irritation, and there are reports of transient irritation of the throat and trachea with the use of smoke-free nicotine cigarettes. These adverse respiratory effects may be due to activation of nAChRs in epithelial cells of the nose and trachea. PMID- 12537064 TI - Validation assessment of the Chinese-version Chronic Ear Survey: a comparison between data from English and Chinese versions. AB - The Chronic Ear Survey (CES) is a valid, disease-specific measure for the evaluation of health status and treatment effectiveness for adults with chronic suppurative otitis media (CSOM). This study compares the validation properties of the English and Chinese versions of the CES. The CES was translated into Mandarin Chinese by means of a parallel model. The Chinese version of the CES (CCES) was administered to 103 patients in a prospective manner, then was validated according to established criteria for reliability, validity, and longitudinal sensitivity. The CCES demonstrated good test-retest reliability and internal consistency (Cronbach's a = 0.81). The CCES significantly correlated with the Mandarin Chinese (Taiwan) version of the generic 36-Item Short-Form Health Survey (TSF-36). The standardized response mean for the CCES total score was 2.1, indicating excellent sensitivity to clinical change. This validation study demonstrated that the performance characteristics of the CCES were equivalent to those of the English-version CES. The CCES is a valid tool for evaluation of adults with CSOM among the Chinese-speaking population. PMID- 12537065 TI - Olfactory changes at threshold and suprathreshold levels following septoplasty with partial inferior turbinectomy. AB - Partial inferior turbinectomy with septoplasty is routinely carried out for airway obstruction. However, its effects on the sense of smell have not been systematically evaluated. The aim of this prospective study was to evaluate the influence of septoplasty with partial inferior turbinectomy on threshold and suprathreshold olfactory acuity. The subjects were 30 patients undergoing septoplasty including partial inferior turbinectomy. Olfactory function was determined by the "Sniffin' Sticks," which allow the assessment of odor thresholds, odor discrimination, and odor identification. The patients rated both olfactory function and nasal airflow using visual analog scales. Nasal airflow was measured by anterior rhinomanometry. Multivariate analyses of variance for repeated measures were used to analyze the results before and after surgery (mean interval, 9.1 weeks). After operation, 87% of the patients had increased airflow, 80% had improved olfactory function in terms of odor identification, and in 70% odor discrimination was found to be improved - but only 54% had improved olfactory function in terms of odor thresholds. Surgery increased ratings of nasal airflow in 93%, and those of olfactory function in 77% (p < .001). Similarly, bilateral inspiratory nasal flow increased (p < .001) and olfactory function was improved (p < .001) after surgical treatment. However, this increase was most pronounced for suprathreshold tests, while it was moderate for odor thresholds (interaction "surgery" x "olfactory test," p = .001). The present investigation suggests that septoplasty in combination with inferior turbinectomy has a beneficial effect on olfaction, mainly on suprathreshold olfactory functions. This effect may be partly due to interactions between the increased perception of nasal airflow and cognitive factors involved in olfactory sensitivity. According to the present results and data from the literature, a moderate decrease of olfactory function appears to occur in as many as 20% of patients. However, anosmia seems to be an extremely rare complication of septoplasty and partial turbinectomy. PMID- 12537066 TI - Coccidioidomycosis of the head and neck. PMID- 12537067 TI - Let's get real. The nether side revealed. PMID- 12537068 TI - Listening to nurses' moral voices: building a quality health care environment. AB - In this paper we describe a research project in nursing ethics aimed at exploring the meaning of ethics for nurses providing direct care with clients. This was a practice-based project in which participants who were staff nurses, nurses in advanced practice, and students in nursing were asked to tell us (or describe to us) how they thought about ethics in their practice, and what ethical practice meant to them. We then undertook to analyze, describe and understand the enactment of ethical practice, the opportunities for and barriers to such enactment, as well as the resources nurses need for ethical practice. We drew out implications of these findings for nursing leaders. We identified practice realities that create a climate for ethical or moral distress, and the way in which nurses attempt to maintain their moral agency. Practice realities included nurses' ethical concerns about policies guiding care; the financial, human and temporal resources available for care; and the power and conflicting loyalties nurses encounter inproviding good care. Maintaining moral agency involved use of a variety of ethical resources and the identification of resources needed to provide good care, as well as the processes used to enact moral agency. Nurse leaders are also moral agents. Important implications of these findings for nursing leaders are that they need moral courage to be self-reflective, to name their own moral distress, and to act so that their nursing staff are able to be moral agents. Nurse leaders need to be the moral compass for nurses, using their power as a positive force to promote, provide and sustain quality practice environments for safe, competent and ethical practice. PMID- 12537069 TI - Conflict in healthcare organizations. AB - Healthcare organizations struggle with internal and external causes of conflict. Successful organizations evaluate and retool existing conflict management systems to constructively, cost-effectively and practically control the negative impact. PMID- 12537070 TI - A multi-disciplinary professional practice model: supporting autonomy and accountability in program-based structure. AB - This article addresses the challenge of maintaining high quality patient care in a large, diverse health care organization as it merged and moved from a functional to a program-based management structure. The article describes the Professional Practice Model (PPM) which was developed to address the professional issues inherent in a program-based management structure. The operationalization of the PPM, including the committee structures which were put in place to support it, are discussed. The final section of the article highlights the strengths and challenges which have arisen through the development and implementation of the Professional Practice Model. PMID- 12537071 TI - Canadian Forces seek out civilian nurses for Case Managers. AB - This article describes a novel case management program implemented by the Canadian Forces Health Services to care for its ill and injured members. A brief overview of the military environment is followed by the reasons why the military looked to the civilian sector and selected case management as a strategy for its continuity of care issues. Principles guiding the design and operations of the program are highlighted along with a description of the core case management activities. Staff roles are outlined including the reasoning behind hiring baccalaureate prepared civilian nurses as Case Managers. The article ends with a description of its current status and notes that preliminary member satisfaction findings demonstrate that nurses are making a positive difference in lives of soldiers that are ill or injured. PMID- 12537072 TI - Harrowing experience of attempting to express our concerns, observations, or symptoms, to Dr. God. PMID- 12537073 TI - Older women's experiences with chronic pain: daily challenges and self-care practices. AB - The subjective nature of chronic pain leads to challenges in describing, treating, and accommodating pain. Twenty older women, who engaged in focus groups, and eight of whom also participated in individual interviews, discussed the influence of chronic pain on their perception of self as well as daily functioning. Managing pain forced many of the women to withdraw from social activities that they valued and interfered with sleep patterns, household tasks and recreational activities. Varied acceptance by social network members added to the challenge of adjusting to complications associated with pain. Self-care practices that supported independence contributed to women's perceived success in daily functioning. PMID- 12537075 TI - Retirement for women: the impact of employment. AB - This study compares the retirement experiences of professional and nonprofessional women. The analysis is based on 48 interviews with women ranging in age from 63-83 years. Participants provided personal and occupational histories and described their retirement decisions, transitions, and lives in retirement. The women's retirement experiences differed in five areas: (1) attachment to work, (2) professional identity, (3) social contacts, (4) family roles/obligations, and (5) community involvement. Findings indicate investment in the work role does affect how women transition to retirement as well as how they structure their time in retirement. PMID- 12537074 TI - Gender and racial inequities in retirement resources. AB - Two waves of a Social Security Beneficiary survey were analyzed to consider differences in the retirement resources of women and men based on marital status and race/ethnicity. Despite increased workforce participation the economic situation of single women, including white women, worsened over time. A bifurcation in retirement resources was found, with men relying more on private income sources and women depending more on Social Security. Current retirement policies based on privatization will continue to adversely impact women who work at low-paying jobs, receive lower wages, and live longer than men. PMID- 12537076 TI - Looking back . . . looking forward: addressing the lives of lesbians 55 and older. AB - This study explored the life course experiences of older lesbians and their concerns and needs as they age. Interviews with sixty-two lesbians, age 55 and older, living in the three west coast states, revealed common themes related to coming out, family and partner relationships, sources of support, aging-related concerns, personal accomplishments, and hopes for the future. Implications for research with older lesbians and for appropriate social work practice and service delivery with this aging population are discussed. PMID- 12537077 TI - "My mind's made up": assumptions and decision-making in accounts of caregiving women. AB - In this qualitative study, rural women caregivers for elderly relatives or friends were interviewed about their decision-making. Women deliberating decisions differed from women implementing decisions in how they perceived their caregiving tasks and in their experience of stress. The difference in deliberative and implemental accounts suggests that mindset is a mediating factor for the women in the present study. Women in the process of implementing decisions regarding caregiving described their experiences in a more positive light and reported less stress. Deliberating decisions regarding caregiving, however, was likely to color the experience of caregiving, leaving the caregiver feeling vulnerable, doubtful about herself, and more stressed. PMID- 12537078 TI - Variations in nutritional risk among black and white women who receive home delivered meals. AB - The maintenance of good nutritional status is a cornerstone of health, function, and quality of life. To assess indicators and correlates of high levels of nutritional risk, sociodemographic, functional status, and nutritional data were collected from a diverse sample of 729 older women. Results from ordered logit analysis indicated that being Black, having an income < or = 125% of the federal poverty level, living alone, and being in the younger-old age group (ages 60 to 74 years) were associated with moderately high and very high levels of nutritional risk. An examination of individual and multiple nutritional risk indicators suggested a heightened vulnerability of Black women to inadequate dietary intake, even net of income and other influential variables. With demographic projections of an increasingly numerous and diverse older population who may face health and nutritional challenges, researchers must strengthen efforts to assist policymakers and service providers to properly target nutrition services. PMID- 12537079 TI - Historical, economic, and political contexts of aging in African America. AB - This article is an attempt to present some of the historical and political contexts of older women in general and to communicate pieces of the lives of African American older women, in particular. It presents some of the present day distortions as well as a sense of the historical, economic, and political realities of older African American lives, while trying to limit the use of the usual comparison approach to this topic. PMID- 12537080 TI - Influence of yard work and weight training on bone mineral density among older U.S. women. AB - The purpose of this study was to determine the influence of type of physical activity on bone mineral density among older U.S. women. Findings from the present study indicate that jogging, swimming and calisthenics were weak predictors for high bone density values. Bicycling, aerobics, walking and dancing were moderate predictors for positive bone density. Yard work and weight training were strong and independent predictors for positive bone density. PMID- 12537081 TI - Health-related interactions and the self in marriage. AB - This study investigated the association between self processes and married partners' (N = 59 couples) perspectives of their health-related social interactions. Findings revealed that wives' self processes were associated with their social behavior from the perspective of each partner. The wives' self processes differentially predicted wives' and husbands' perspectives of their interactions, however. These findings demonstrate that wives' future expectations for the health of their husband, as well as for their role in maintaining his future health, motivate their current social behavior to promote the positive health lifestyle behaviors of their husband. PMID- 12537082 TI - Taking charge of self: breast health practices of older rural women. AB - The purpose of this grounded theory study was to explore breast health practices of older Vermont women residing in rural communities. Although the three components of breast health-mammography, clinical breast exam, and self-breast exam-are recommended for women 40 years and over, minimal research has empirically analyzed the breast health practices of healthy women to ascertain if, in fact, these procedures are followed, and if so, what the motivation is for doing so. Twelve women, 50-64 years, participated in face-to-face, audiotaped interviews. Data analysis, including line-by-line and constant comparative approaches, occurred concurrently with data collection. Taking Charge of Self, the generated theory, describes participants' engagement in a process of learning how to take charge of their lives. The analysis indicates that health care providers have a powerful role in the lives of women in this age group. With an increasing emphasis on health promotion and disease prevention, health care providers are positioned to cultivate and support women's development of the personal or internal motivation for health and well-being. PMID- 12537083 TI - Midlife myths and realities: women reflect on their experiences. AB - This was one in a series of studies whose goal was creation of a rich, deep description of women's midlife psychological development. All studies in the series used qualitative approaches of unstructured focus groups followed by grounded-theory data analysis. The participants were a culturally and ethnically mixed group of 11 women between ages 40 and 60 who met in Manhattan and discussed their lives, feelings, behaviors, and perspectives during this important period. Data analysis led to a three-part developmental model describing women's midlife experiences. Data from this group of participants offered particularly rich descriptions of how they reacted to social stereotypes about midlife. PMID- 12537084 TI - Drug-eluting stents to prevent reblockage of coronary arteries. AB - Restenosis limits the success of percutaneous transluminal coronary interventions. Coronary artery stenting decreases restenosis, improves outcomes, and is currently the most commonly used percutaneous coronary intervention in the United States. However, in-stent restenosis continues to occur at an unacceptable rate. In-stent restenosis is a neointimal hyperplastic response resulting primarily from vascular smooth muscle cell proliferation. Treatment with anti proliferative agents presents a logical approach to eradicating restenosis, however, these drugs are highly toxic. Coating stents with anti-proliferative agents allows local delivery of high doses and avoids systemic side effects. In 2001, the results of two clinical trials, RAVEL and ELUTES, using sirolimus- and paclitaxil-coated stents demonstrated nearly complete elimination of in-stent restenosis. These dramatic results represent a tremendous advance in the treatment of coronary heart disease. PMID- 12537085 TI - Current and potential applications of left ventricular assist devices. AB - The syndrome of heart failure is epidemic, causing increased hospital admissions, poor survival rates, and a dismal quality of life. Treatment choices include pharmacologic, psychological, and surgical interventions. Recent data suggest that mechanical heart pumps, or assist devices, are viable additions or alternatives to transplantation in management of advanced heart failure. A ventricular assist device (VAD) can support the circulation when the natural heart is unable to maintain adequate perfusion. Although its use continues to be explored, VADs can be used in three ways: as a bridge to transplantation, as a destination therapy (or implanted permanently), or as a bridge to recovery. As VAD science advances, nurses will needs to understand its practice implications. PMID- 12537086 TI - The AbioCor totally implantable replacement heart. AB - An artificial heart with adequate circulatory support and an acceptable quality of life remains one of the holy grails of heart failure medicine and surgery. The totally implantable AbioCor is powered electrically via an external power source and has no skin-piercing cables. To date, seven critically ill patients with end stage heart failure have been implanted with it. Four patients survived beyond 2 months, and two patients were discharged from the hospital. Both enjoyed improved quality of life with frequent social excursions; another patient is about to be discharged. While three patients died, early trials suggest that this device holds promise. PMID- 12537087 TI - Cardiovascular tissue engineering: constructing living tissue cardiac valves and blood vessels using bone marrow, umbilical cord blood, and peripheral blood cells. AB - Although atherosclerosis and valvular heart disease are among the leading causes of morbidity and mortality in developed nations, the substitute blood vessels and heart valves currently available all have significant limitations. During the past 10 years, a new field called tissue engineering has emerged, and several research groups are focusing their efforts on constructing living tissue replacement blood vessels and heart valves. In 2001 several exciting developments occurred with the use of progenitor and stem cells. This article introduces the essential concepts of cardiovascular tissue engineering, reviews achievements in the field, discusses the basic developmental biology of heart valves and blood vessels, and summarizes the 2001 research on progenitor and stem cells. PMID- 12537088 TI - Gene therapy with vascular endothelial growth factor reduces angina. AB - A placebo-controlled, double-blind, randomized study found that subjects randomized to the vascular endothelial growth factor (VEGF) gene-receiving treatment group showed a greater level of angina reduction in comparison to control subjects who received saline as a placebo. These data provide hope for a new treatment option for those who are not candidates for invasive therapeutic procedures and are refractory to medical therapy for angina. Furthermore, the findings are important to the areas of therapeutic angiogenesis and gene therapy as a whole. This article discusses VEGF and its brief history as a form of gene therapy in the context of the VEGF gene therapy trial that the American Heart Association has recognized as one of the top 10 scientific advances of 2001. PMID- 12537089 TI - Cholesterol-lowering drugs bring benefits to high-risk populations even when LDL is normal. AB - The Heart Protection Study (HPS) results were presented shortly after the National Cholesterol Education Program released the third report of clinical practice guidelines for the evaluation and management of elevated cholesterol in adults. Both the guidelines and the HPS results support an aggressive approach to managing high-risk individuals. The HPS showed that cardiovascular events and mortality were reduced in high-risk patients taking simvastatin regardless of baseline low-density lipoprotein (LDL) cholesterol levels. Some of these patients would not have met criteria for drug treatment according to the guidelines. The conclusion was that all high-risk patients with LDL > or = 100 mg/dL should be treated with drug therapy with the goal of reducing LDL to < 100 mg/dL. PMID- 12537090 TI - Genetic predictors of cardiovascular disease: the use of chip technology. AB - Cardiovascular disease, a primary cause of mortality in the United States, has a complex pathologic process involving many genes. The high-throughput technology (microarray or "DNA chip") used to decipher the human genome is now being employed to identify key genes in its development. A study focusing on candidate genes associated with premature cardiovascular disease discovered that missense variations in the thrombospondin 1 and 4 genes were associated with premature coronary artery disease, while a mutation in the non-coding region of a thrombospondin 2 gene imparts protection from developing heart disease. Although the clinical implications of microarray technology are still under investigation, this research may lead to a diagnostic test to determine a patient's risk for developing heart disease. PMID- 12537091 TI - Cell transplants offer promise for stroke recovery. AB - Cell transplantation is an experimental approach to restore brain function in neurodegenerative disorders such as Parkinson's and Huntington's disease. Transplantation also represents a possible strategy to repair the brain after a stroke. Various cell types are under investigation in experimental stroke studies. This review discusses the different graft sources and presents preliminary data on the transplantation of neural progenitor cells after stroke in rats. Following transplantation, progenitor cells proliferated and differentiated into all the different brain cell types, including neurons, and they repopulated the ischemic infarct. These results suggest that cell transplantation may serve as a future restorative therapy for stroke and other neurologic disorders such as Parkinson's disease, Alzheimer's disease, trauma, and multiple sclerosis. PMID- 12537092 TI - Behavior and biology: the prevention of type 2 diabetes. AB - Type 2 diabetes mellitus (DM), characterized by insulin resistance and a beta cell secretory defect, appears to result from a number of gene and environmental interactions. There are marked differences in the phenotypic expression of type 2 DM with individuals exhibiting varying levels of insulin resistance and impairments in insulin secretion. Study results indicate that a number of healthy lifestyle behaviors, such as increased physical activity and reduced intake of dietary fat, are associated with decreased development of type 2 DM. This article explores the genetic and environmental factors associated with the development of type 2 DM along with the role of lifestyle modifications in the prevention of this disease. PMID- 12537093 TI - Passive smoking and vascular disease. AB - Passive smoking, or environmental tobacco smoke, is a causative factor in cardiovascular disease. A 30-minute passive smoking exposure was found to affect coronary flow velocity reserve in nonsmokers, indicating endothelial dysfunction in coronary circulation. This article summarizes empirical work on passive smoking and heart disease. Clinically relevant findings include a dose-response relationship between passive smoking exposure and heart disease and partial reversibility of physical effects after eliminating passive smoking exposure. Appropriate assessment of passive smoking exposure in a variety of settings is warranted, as well as recommendations to avoid such exposure. Policy-based public health initiatives to eliminate passive smoking in the workplace and other public areas are needed. PMID- 12537094 TI - Introduction of Mark E. Siddall as the Henry Baldwin Ward Medalist for 2002. PMID- 12537095 TI - Resolution and independence--acceptance of the 2002 Henry Baldwin Ward medal. PMID- 12537096 TI - Introduction of J. Richard Seed, recipient of the 2002 Meritorious and Distinguished Service Award. PMID- 12537097 TI - Intensity-dependent alteration of minnow (Pimephales promelas) behavior by a brain-encysting trematode. AB - We examined the relationship between the numbers of brain-encysting trematodes (Ornithodiplostomum ptychocheilus) and the magnitude of altered behaviors in fathead minnows (Pimephales promelas). Because cysts develop within a brain region that integrates visual stimuli with motor response. we evaluated the standard optomotor response (OMR). Monitoring this task involved recording the time minnows spent following a spinning drum, on which alternating black and white stripes had been painted. Minnows were exposed to 0, 5, 20, 120, and 300 cercariae and then their OMR was evaluated at 2-wk postinfection. Surprisingly, only minnows that had high numbers of parasites (155 +/- 31 worms/fish) or low numbers of parasites (3 +/- 3 worms/ fish) differed significantly in their optomotor performance compared with controls. Reduced OMR of heavily infected minnows was positively correlated with reduction in minnow activity. In contrast, reduced OMR in lightly infected minnows was independent of host activity and was likely associated with the rapid development of parasite larvae within the optic tecta. The nonlinear relationship between parasite intensity and effect on host behavior was consistent with an earlier study, but the underlying mechanisms producing this pattern are unknown. PMID- 12537098 TI - Spatial and temporal variations in the trematode component community of the mudsnail Hydrobia ventrosa in relation to the occurrence of waterfowl as definitive hosts. AB - The component community of larval trematodes infecting the mudsnail Hydrobia ventrosa (Montagu) was examined in coastal lagoons of the southern Baltic Sea among different host subpopulations in relation to the structure of the waterfowl community. The 10 trematode species observed represent the families Notocotylidae (1), Echinostomatidae (1 or 2), Heterophyidae (2). Monorchidae (1). Microphallidae (3 or 4), Psilostomatidae (1), and Hemiuridae (1). Eight of these species infect waterfowl as adults. The structure of the trematode communities was similar between sampling sites. Seven trematode taxa were commonly found at all sampling sites. Prevalence values of the 6 most abundant taxa, which infect birds as final hosts, were significantly different between neither sampling sites nor across year. Overall trematode prevalence in H. ventrosa fluctuated seasonally. Prevalence usually peaked in summer between July and September or October. Low prevalences were observed in late winter and early spring. In contrast, the seasonal maximum in waterfowl numbers differed between areas because of significant spatial differences in the bird community structure. The species composition of the component trematode community of H. ventrosa in the coastal lagoons of the southern Baltic Sea is more or less independent of the species composition of the waterfowl community. This independence presumably results from the lack of host specificity in most of the observed trematode species. Otherwise, the low host specificity in combination with the enormous waterfowl diversity in the coastal lagoons might explain the stability of the prevalence pattern of the component trematode community. PMID- 12537099 TI - Metazoan parasite infracommunities of Florida pompano (Trachinotus carolinus) from the coast of the Yucatan Peninsula, Mexico. AB - Metazoan parasite infracommunities of the Florida pompano (Trachinotus carolinus) were studied in terms of species composition, species richness, diversity, numerical dominance, and similarity. Seventy-five fishes were collected from 4 localities along the Yucatan Peninsula coast and 24 parasite species recovered. Most were digeneans (8 species) and nematodes (7). Other species were monogeneans (3). aspidogastreans (2), cestodes (1), acanthocephalans (1), and crustaceans (2). Only 4 species were common in at least I locality. Mean values for species richness, abundance, diversity, numerical dominance, and similarity in total (all species in the individual fish), gastrointestinal, and ectoparasite infracommunities were within ranges observed for most helminth infracommunities of marine fishes from temperate and tropical latitudes. These infracommunities had low species richness, abundance, diversity, and predictability (except ectoparasite infracommunities) and high dominance. Within the predictable element (common species), the specialist monogenean Pseudobicotylophora atlantica was the main reason for the increase in predictability because it was the only common species at all 4 localities. Host feeding habits, the distribution of intermediate hosts and infective stages, the local species pool, and a phylogenetic component seem to be determining the characteristics of these metazoan parasite infracommunities. PMID- 12537100 TI - Neospora caninum in vitro: evidence that the destiny of a parasitophorous vacuole depends on the phenotype of the progenitor zoite. AB - We previously reported that Neospora caninum can be induced to express BAGI, a bradyzoite antigen, within 3 days of culture under stress conditions. The main goals of the present experiment were to increase the expression of BAGI in vitro (in part by extending cultures for 9 days), to observe parasitophorous vacuoles at various points of stage differentiation, and to test the ability of organisms produced in vitro to function like mature bradyzoites. Expression of BAG1 and of a tachyzoite antigen (NcSAGI) was monitored using a double-label immunofluorescence assay. For the purpose of this study, organisms expressing NcSAG1 were designated as tachyzoites, those expressing BAG1 were designated as bradyzoites, and those expressing both antigens were designated as intermediate zoites. The greatest percentage of intermediate zoites and bradyzoites (14%) occurred in bovine monocytes maintained for 9 days. These bradyzoites did not appear to be functionally mature; they did not induce patent infections in dogs. in contrast to bradyzoites that were produced in chronically infected mice. In vitro, large parasitophorous vacuoles contained either a pure population of tachyzoites or a mixture of tachyzoites and intermediate zoites, which is indicative of asynchronous stage conversion of organisms within a vacuole. Bradyzoites were first observed within small vacuoles on day 6. and bradyzoites never shared vacuoles with tachyzoites. This finding suggests that vacuoles containing bradyzoites may develop only if the cell is invaded by a zoite that has already begun bradyzoite differentiation. An alternative possibility is that cysts may develop if the establishing tachyzoite undergoes bradyzoite differentiation before multiplying. Cysts do not appear to arise from transformation of tachyzoites within large parasitophorous vacuoles. PMID- 12537101 TI - Structural and biochemical alterations in Giardia lamblia cysts exposed to ozone. AB - Because of its efficacy in inactivating waterborne protozoan cysts and oocysts, ozone is frequently used for disinfection of drinking water. The effect of ozone on cysts of Giardia lamblia was investigated in gerbils (Meriones unguiculatus), using an infectivity assay by scanning electron microscopy, immunoblotting, and flow cytometry. Cysts recovered from experimentally infected gerbils were exposed to an initial ozone concentration of 1.5 mg/L for 0, 30, 60, and 120 sec. This treatment resulted in a dose-dependent reduction in cysts concentration, loss of infectivity in gerbils, and profound structural modifications to the cyst wall. Exposure for 60 sec or longer resulted in extensive protein degradation and in the disappearance of a cyst wall and a trophozoite antigen. PMID- 12537102 TI - Relationship between long-term resistance to Trypanosoma cruzi and latent infection, examined by antibody production and polymerase chain reaction in mice. AB - Trypanosoma cruzi infections persist for the lifetime of humans and laboratory animals as either latent or pathogenic parasitism. Mice inoculated with a nonpathogenic, attenuated strain (TCC) display resistance against virulent challenge, with a strong control of parasitemia and protection against tissue lesions for more than 12 mo. Three main approaches were used to test whether protection by TCC inocula is based on a latent infection or on a "sterile" immunological memory: curative Benznidazole (Bzl) treatment, serological reactions, and detection of infection by polymerase chain reaction (PCR). If resistance is maintained in the absence of infection, it should not be reduced by Bzl treatment and TCC-inoculated animals should not maintain long-term serological or PCR reactivity. The Bzl treatment after TCC inoculations did not reduce, after periods of up to 420 days, TCC-induced resistance to challenge. But TCC inocula given during Bzl treatment conferred short-term, but not long-term. protection. Maintenance of high antibody levels and protection were better in the virulent Tulahuen (TUL) strain than in the attenuated TCC strain infections, and trypomastigote inocula of either strain were better inducers of antibodies and resistance than epimastigotes. PCR detection of T. cruzi DNA was positive in almost all TUL strain-inoculated animals and negative in immunocompetent animals inoculated with TCC epimastigotes, although high numbers of TCC trypomastigotes produced persistent PCR signals of infection in newborn BALB mice. Thus, 2 polar models were developed, where latent infection by TCC was either demonstrated or excluded. In both, resistance to virulent challenge was maintained during long periods. But late declination of antibody titers (>200 days) and resistance to challenge (>350 days) was observed in animals displaying clearance of all signals of infection. PMID- 12537103 TI - Characterization of Neospora caninum protease, NcSUB1 (NC-P65), with rabbit anti N54. AB - NcSUB1 (formerly known as NC-p65) is the first molecularly described proteolytic enzyme of the intracellular protozoan parasite Neospora caninum. This report describes the characterization of a rabbit anti-N54, which is an antiserum generated against an internal fragment of NcSUB1 (amino acids 649-783). In immunofluorescence studies rabbit and-N54 labeled the apical end of the fixed parasite. By immuno-gold electron microscopy, the antibody bound primarily to the microneme organelles of the parasite. Analysis of secreted parasitic proteins indicated that a protein of molecular weight 65 kDa (reduced) or 55 kDa (nonreduced) was recognized bythe antibody. The same secreted proteins were affinity purified with rabbit anti-N54-coupled resins and were shown to contain major proteolytic activity by zymography. Thus, rabbit anti-N54 is the first antibody developed for N. caninum that binds to themicroneme proteins and recognizes a major secreted enzyme. PMID- 12537104 TI - Experimental infection of sheep with Neospora caninum oocysts. AB - The purpose of the present study was to investigate the potential of Neospora caninum oocysts to infect sheep and determine whether N. caninum DNA could be detected by polymerase chain reaction (PCR) assay in blood and brain of sheep after oocyst inoculation. Six ewes were inoculated per os with 10(4) N. caninum oocysts, whereas 2 ewes served as uninoculated controls. All sheep were bled weekly for 7 wk after inoculation. Blood was analyzed for the presence of N. caninum DNA by 2 different PCR assays, as well as for the presence of antibodies to recombinant and native N. caninum antigens. Neospora caninum DNA was detected in 2 sheep as early as 7 days after oocyst inoculation (DAOI). All 6 sheep were PCR positive by 32 days and remained positive until the end of the study at 49 DAOI. Aside from 1 ewe, all sheep inoculated with N. caninum oocysts contained detectable N. caninum DNA in the brain tissue collected at 49 DAOI. Unlike with PCR, no lesion or parasite was detected by immunohistochemistry. Antibodies were detected by enzyme-linked immunosorbent assay, Neospora agglutination test, or immunoblotting to either native or recombinant N. caninum antigens in sheep inoculated with oocysts. PMID- 12537105 TI - In vitro and in vivo encystment of the cercariae of Echinostoma caproni. AB - In vitro and in vivo studies were conducted on the cercariae of Echinostoma caproni. Of the 15 media tried, 2 resulted in effective in vitro encystment in petri dish cultures maintained at 23 +/- 1 C. They were a Locke's--artificial springwater (ASW) (1:1) medium (67% encystment) and a Biomphalaria glabrata embryonic cell line medium (23% encystment). To obtain large numbers of in vitro- formed cysts, finger bowl cultures containing 40 ml of the Locke's-ASW (1:1) medium were used at 23 +/- 1 C. Of 3,000 cercariae tested, 1,890 (63%) were encysted in this medium by 48 hr. Most of these cysts looked similar to those formed in vivo, although some showed abnormalities in the outer cyst wall and other malformations. A total of 200 in vitro-formed cysts treated in an alkaline trypsin-bile salts (TB) medium for 2 hr at 41 C showed 94% excystation. In vitro formed cysts fed to mice produced ovigerous adults within 2 wk postinfection (PI). Eggs from these worms gave rise to miracidia that produced patent intramolluscan infections in B. glabrata snails. In vivo encystment was studied in lab-raised juvenile Helisoma trivolvis (Colorado strain) snails, 1-3 mm in shell diameter. From 6 to 24 hr PI, 93-100% of the cercariae were recovered as metacercarial cysts in the snail tissue. Treatment of these cysts in the TB medium resulted in 96% excystation within 2 hr at 41 C. PMID- 12537106 TI - Gastrointestinal helminths of ponies in Louisiana: a comparison of species currently prevalent with those present 20 years ago. AB - A survey in Louisiana of gastrointestinal helminths recovered at necropsy from 117 ponies with minimal exposure to anthelmintics between 1989 and 2000 is compared with a survey conducted 20 yr earlier in the same region. An objective of this study was to determine whether species diversity has been affected by the advent and use of the macrocyclic lactone (ML) parasiticides and by the increased anthelmintic pressure on the helminth species infecting the general equine population. Twenty-six cyathostome species and 8 strongyle species were recovered. Two cyathostome species that were not found before, Cylicostephanus asymetricus and C. bidentatus, and 1 species of large strongyle, Oesophagodontus robustus, were added to the list of species found in Louisiana. All cyathostome and large strongyle species found previously were still present. But prevalences and intensities were significantly reduced for almost all large and small strongyle species. Prevalences and intensities of Oxyuris equi adults and larvae were reduced, whereas the prevalence of Parascaris equorum remained constant. The tapeworm Paranoplocephala mamillana was added to the list of parasite species found in Louisiana. Anoplocephala perfoliata remained the most common cestode. This species was found at the same level of intensity but increased slightly in prevalence. Anoplocephala magna was found less frequently than previously. The overall diversity of species remained reatively unchanged. The reasons for the differences in intensity and prevalence of strongyles between these 2 periods are unknown but might be related to the development and use of the broad-spectrum ML anthelmintics in the intervening period, a difference in the population of equids surveyed, different techniques used to identify the parasites, or differences in numbers of parasites identified (or to all). PMID- 12537107 TI - Infection patterns of Tylodelphys barilochensis and T. crubensis (Trematoda: Diplostomatidae) metacercariae in Galaxias maculatus (Osmeriformes: Galaxiidae) from two Patagonian lakes and observations on their geographical distribution in the southern Andean region, Argentina. AB - In the Patagonian Andean region, 2 species of diplostomatids parasitize the brains of Galaxias maculatus. The purpose of this study was to evaluate seasonal variation, spatial variation, and association with host age in the transmission of Tylodelphys barilochensis and T. crubensis in several oligotrophic lakes in Argentinian Patagonia. Fishes were captured monthly in Lake Gutierrez and bimonthly in Lake Escondido. One summer or autumn sample was also taken in several other Patagonian lakes. Infection parameters were calculated and compared using nonparametric tests. The 2 species co-occurred in most of the sampled lakes, with high values of prevalence and mean intensity. In Lake Gutierrez and Lake Escondido, the intensity of both diplostomatid species did not show significant differences between sexes and co-varied with host length. All age classes were infected; maximum prevalence values were reached before maximum mean intensity values in the 1-yr age class. A seasonal pattern of prevalence and mean intensity of the 2 parasite species with autumn mean intensity values differing significantly from those of the other seasons was evident only in Lake Gutierrez. PMID- 12537108 TI - Parasitic helminths of red-bellied woodpeckers (Melanerpes carolinus) from the Apalachicola National Forest in Florida. AB - Seventy-four red-bellied woodpeckers (Melanerpes carolinus) from the Apalachicola National Forest (30 degrees 10'N, 84 degrees 40'W) in northwest Florida were examined for helminths. The most prevalent parasites were the nematode Aproctella stoddardi (11%) and the acanthocephalan Mediorhynchus centurorum (11%). New host records include Pseudaprocta samueli, A. stoddardi, Tridentocapillaria tridens, Diplotriaena americana, Dispharynx nasuta, Procyrnea pileata, Orthoskrjabinia rostellata, and Brachylaima fuscatum. The helminth fauna was characterized by low prevalences and intensities of infection and low numbers of species per bird (1.2). The frequency of prescribed burning and habitat understory flora composition did not influence the prevalences or intensities of helminths in red bellied woodpeckers collected from 2 similar but differently managed sites within the forest. PMID- 12537109 TI - Experimental life cycle of Lagochilascaris major leiper, 1910 (Nematoda: Ascarididae) in cats (Felis domesticus). AB - The life cycle of Lagochilascaris major was studied using eggs collected from a natural clinical case in a domestic cat. Twenty-seven white mice (Mus musculaus), 5 hamsters (Mesocricetus auratus), and 1 vesper mouse (Calomys callosus) were orally inoculated with 800-1,300 embryonated eggs. When examined from 73 to 246 days postinoculation (PI), encysted third-stage larvae were seen in skeletal muscles and less frequently in connective tissue, liver, and lungs. Twenty-two of the 23 cats orally inoculated with 40-430 encysted larvae from these rodents, and necropsied from 1 hr to 185 days PI, became infected. Third-stage larvae were located in the stomach, esophagus, and oropharynx from 1 to 24 hr PI. At 48 hr, larvae, from mainly the fourth stage, were only found, unilaterally or bilaterally, inside a "sac" in the region of the semilunar fold of the palatine tonsil at the base of the tongue. Adult worms were found in this location from 10 to 175 days PI. No fistulated abscess to the outside medium was found. Adult worms were also found in the middle ears of 2 cats showing purulent otitis. Eggs in the ear secretion were under different stages of development. Eggs in feces were first observed on days 14 and 15 PI, and 1 cat shed them until 178 days PI. Six infected cats were treated with fenbendazole at 50 mg/kg of body weight for 3 consecutive days, eliminating all the parasites present in the tonsils. The drug was not effective against the parasites present in the middle ear. No stage of the parasite was found in the tissues of 5 cats given 4,000-5,200 eggs orally and examined after 19 and 50 days PI. This indicates that the life cycle of L. major requires an obligate paratenic host and is characterized by heteroxenic cycle. PMID- 12537110 TI - Life cycle of Sarcocystis neurona in its natural intermediate host, the raccoon, Procyon lotor. AB - Sarcocystis neurona causes encephalomyelitis in many species of mammals and is the most important cause of neurologic disease in the horse. Its complete life cycle is unknown, particularly its development and localization in the intermediate host. Recently, the raccoon (Procyon lotor) was recognized as a natural intermediate host of S. neurona. In the present study, migration and development of S. neurona was studied in 10 raccoons that were fed S. neurona sporocysts from experimentally infected opossums; 4 raccoons served as controls. Raccoons were examined at necropsy 1, 3, 5, 7, 10, 14, 15, 22, 37, and 77 days after feeding on sporocysts (DAFS). Tissue sections of most of the organs were studied histologically and reacted with anti-S. neurona-specific polyclonal rabbit serum in an immunohistochemical test. Parasitemia was demonstrated in peripheral blood of raccoons 3 and 5 DAFS. Individual zoites were seen in histologic sections of intestines of raccoons euthanized 1, 3, and 5 DAFS. Schizonts and merozoites were seen in many tissues 7 to 22 DAFS, particularly in the brain. Sarcocysts were seen in raccoons killed 22 DAFS. Sarcocysts at 22 DAFS were immature and seen only in skeletal muscle. Mature sarcocysts were seen in all skeletal samples, particularly in the tongue of the raccoon 77 DAFS; these sarcocysts were infective to laboratory-raised opossums. This is the first report of the complete development of S. neurona schizonts and sarcocysts in a natural intermediate host. PMID- 12537111 TI - Improved production of Neospora caninum oocysts, cyclical oral transmission between dogs and cattle, and in vitro isolation from oocysts. AB - Scarce information is available about Neospora caninum oocysts and sporozoites, in part because only small numbers of oocysts have typically been produced by experimentally infected dogs. We hypothesized that I reason for low experimental production of oocysts is that dogs have been fed tissues from experimentally infected mice instead of tissues from cattle (which are natural intermediate hosts of N. caninum). In this study, 9 dogs were fed tissues from N. caninum infected calves, and oocyst production was compared with 6 dogs that were fed infected mouse carcasses. The number of oocysts produced by dogs that ingested infected calf tissues (mean = 160,700) was significantly greater (P = 0.03) than the number of oocysts shed by dogs that ingested infected mice (mean = 5,400). The second goal of our experiment was to demonstrate cyclical oral transmission of N. caninum between dogs and cattle. As few as 300 oocysts were used to successfully infect calves, and tissues from these calves induced patent infections in 2 of 3 dogs; oocysts from I of these dogs were administered to another calf, and tissues from this calf subsequently induced a third dog to shed oocysts. Oocysts were confirmed to be N. caninum using a species-specific polymerase chain reaction technique. In addition, sporulated oocysts were used to recover N. caninum in vitro after digestion in an acid-pepsin solution and inoculation of cell monolayers. PMID- 12537112 TI - Experimental induction of equine protozoan myeloencephalitis (EPM) in the horse: effect of Sarcocystis neurona sporocyst inoculation dose on the development of clinical neurologic disease. AB - The effect of inoculation dose of Sarcocystis neurona sporocysts on the development of clinical neurologic disease in horses was investigated. Twenty four seronegative weanling horses were subjected to the natural stress of transport and then randomly assigned to 6 treatment groups of 4 horses each. Horses were then immediately inoculated with either 10(2), 10(3), 10(4), 10(5), or 10(6) S. neurona sporocysts or placebo using nasogastric tube and housed indoors. Weekly neurologic examinations were performed by a blinded observer. Blood was collected weekly for antibody determination by Western blot analysis. Cerebrospinal fluid was collected before inoculation and before euthanasia for S. neurona antibody determination. Horses were killed and necropsied between 4 and 5 wk after inoculation. Differences were detected among dose groups based on seroconversion times, severity of clinical neurologic signs, and presence of microscopic lesions. Seroconversion of challenged horses was observed as early as 14 days postinfection in the 10(6) sporocyst dose group. Mild to moderate clinical signs of neurologic disease were produced in challenged horses from all groups, with the most consistent signs seen in the 10(6) sporocyst dose group. Histologic lesions suggestive of S. neurona infection were detected in 4 of the 20 horses fed sporocysts. Parasites were not detected in equine tissues by light microscopy, immunohistochemistry, or bioassay in gamma-interferon gene knockout mice. Control horses remained seronegative for the duration of the study and had no histologic evidence of protozoal infection. PMID- 12537113 TI - Two new species of Yorkeria (Tetraphyllidea: Onchobothriidae) from Chiloscyllium punctatum (Elasmobranchii: Hemiscylliidae) in Thailand. AB - Two new species of the tapeworm genus Yorkeria are described from the spiral intestine of a specimen of the Brownbanded bambooshark, Chiloscyllium punctatum, collected from a fish market in Bangsarai, Thailand in 1996. The larger of the 2 new tapeworm species, Y. hilli, possesses fewer proglottids, fewer testes, and is smaller than most of its congeners. The smaller of the 2 new species, Y. kelleyae, conspicuously differs from all of its congeners in its possession of medial hooks that are only slightly larger than the lateral hooks (rather than conspicuously larger). Scanning electron microscopy shows both species to possess elongate filitriches on their distal prehook locular surfaces and elongate filitrichs and spinitrichs on their proximal bothridial, distal posthook locular surfaces, pedicels, and cephalic peduncles. In both species, the spinitrichs on the proximal bothridial surfaces, pedicels, and peduncles are very large and readily visible with light microscopy. Whereas the spinitrichs on these 4 surfaces are bluntly rounded in Y. hilli, they are pointed in Y. kelleyae. Examination of specimens identified as Y. parva, collected by several previous workers from a diversity of hosts and localities, calls into question the conspecificity of at least some of this material. The unusual scolex form suggests that the configuration of the cerebral ganglionic mass in species of Yorkeria may differ from that seen in most other tetraphyllidean taxa. PMID- 12537114 TI - Neodiplostomum leei n. sp. (Digenea: Neodiplostomidae) from chicks infected with metacercariae from the grass snake Rhabdophis tigrina. AB - Neodiplostomula from the European grass snake Rhabdophis tigrina, in the Republic of Korea, were thought to represent 2 species: Pharyngostomum cordatum (large sized neodiplostomula), an intestinal trematode of cats, and Neodiplostomum seoulense (small-sized neodiplostomula), an intestinal trematode of humans and rodents. The present study describes N. leei n. sp. (Digenea: Neodiplostomidae) on the basis of adult flukes recovered from the small intestines of chicks experimentally infected with small-sized neodiplostomula from the grass snake. The new species differs from N. seoulense in terms of the more extensive distribution of vitellaria, the more severe bilobation of 2 testes (each having a dumbbell shape), the absence of a genital cone, and the smaller size of the tribocytic organ and eggs. Neodiplostomula of the new species and N. seoulense are almost indistinguishable. The new species develops into an adult in chicks, whereas N. seoulense matures in mice and rats. Neodiplostomula of the new species migrate to the liver in mice and rats, without maturing. Results show that there are at least 3 species of neodiplostomid trematodes in the grass snake in the Republic of Korea, i.e.. P. cordatum, N. seoulense and N. leei n. sp. PMID- 12537115 TI - Calliobothrium spp. (Eucestoda: Tetraphyllidea: Onchobothriidae) in Mustelus schmitti (Chondrichthyes: Carcharhiniformes) from Argentina and Uruguay. AB - Three species of Calliobothrium inhabit the spiral intestine of Mustelus schmitti in Argentina and Uruguay. Calliobothrium verticillatum australis is redescribed and its taxonomic status modified to species as C. australis. Calliobothrium barbarae n. sp. can be distinguished from all other species of Calliobothrium, which are small bodied, nonlaciniate, and without accessory piece between the bases of axial hook, by worm length, number of segments, cocoon morphology, and hooks shape. Calliobothrium lunae n. sp. is different from other Calliobothrium spp., which are small bodied, nonlaciniate. and have an accessory piece, by the number of segments and testes, hook shape, cocoon morphology, and the presence of ciliumlike projections on the distal surface of muscular pads. Calliobothrium australis is clearly distinguished from other large-bodied, laciniate species of the genus by worm length, number of testes, ovary shape, cocoon morphology, hook shape, and in being hyperapolytic. The oioxenous specificity involving Calliobothrium spp. and Mustelus spp. described by previous authors is confirmed in this study. PMID- 12537116 TI - Haematoloechus danbrooksi n. sp. (Digenea: Plagiorchioidea) from Rana vaillanti from Los Tuxtlas, Veracruz, Mexico. AB - Haematoloechus danbrooksi n. sp. from the lungs of Rana vaillanti in Veracruz state, Mexico, was found. The new species is most similar morphologically to H. medioplexus, H. parviplexus, and H. meridionalis in having a ventral sucker less than half the diameter of the oral sucker. It differs from these species by the extension of the vitellaria, which are shorter in the new species, and in the shape of the tegumental spines, which are blunt in the new species. It differs from all other known species of Haematoloechus in the distribution of the uterine loops that are arranged diagonally and that present several short, extracecal. longitudinal loops in the postacetabular region. The new species shows 1.2% sequence divergence in partial 28S sequence with respect to H. medioplexus, 1.1% with H. parviplexus. and 2.5% with H. meridionalis, sequence divergences complementing the morphological differences. PMID- 12537117 TI - Corynosoma magdaleni in gray seals from the Gulf of Bothnia, with emended descriptions of Corynosoma strumosum and Corynosoma magdaleni. AB - Because of the likelihood that Corynosoma magdaleni Montreuil, 1958. has been confused with C. strumosum (Rudolphi, 1802) in reports of parasites from seals and to clarify its distribution in the Baltic Sea, acanthocephalans from 26 young gray seals from the southwestern Finnish archipelago (western Baltic Sea) were examined. All harbored C. semerme (Forssell. 1904). In addition to C. semerme, 12 had both C. strumosum and C. magdaleni, 3 had only C. strumosum, and 9 had only C. magdaleni. Most anatomical structures of C. strumosum are similar to, but larger than, those of C. magdaleni. The most conspicuous differences are the longer and more robust trunk of C. strumosum, the routinely longer proboscis (mean, 653 vs. 476 microm) and larger proboscis hook (mean, 69 vs. 58 microm) of C. strumosum, and the greater extent of ventral trunk spines (mean, 61 vs. 47%) in C. magdaleni. In addition, C. strumosum consistently possesses 18 longitudinal rows of proboscis hooks, whereas C. magdaleni has 17-23, with 20 being the usual number by far. In seals aged 3.6 mo, on average, C. strumosum was more prevalent and abundant than was C. magdaleni, whereas in seals of age 2.0-3.0 mo the reverse was true, with C. strumosum being nearly absent. These differences might reveal the age-dependent food habits of the very young seals. No site segregation was found between C. strumosum and C. magdaleni in the small intestine, but they clearly segregated from C. semerme, which occurred mainly in the cecum, large intestine, and rectum. All species matured equally well in gray seals, with 62, 67, and 53% of the C. magdaleni, C. strumosum, and C. semerme, respectively, comprising gravid worms (possessing eggs with fully formed acanthors). PMID- 12537118 TI - Description of a new Eimeria sp. and associated lesions in the kidneys of double crested cormorants (Phalocrocorax auritus). AB - A new Eimeria sp. is described as the cause of an outbreak of renal coccidiosis in double-crested cormorants (Phalacrocorax auritus) in Georgia. Sporulated oocysts were spherical to subspherical and measured 16.1 x 16.5 (13.8-18 x 14-19) microm, with an average length-width ratio of 1:1. Oocyst wall was thin (1 microm), greenish, and pitted on the outer surface. Micropyle, micropylar cap, Stieda body, and polar bodies were absent. Small oocyst residuum (4-8 granules) was usually absent but occasionally present. Sporocysts were oval and measured 6.6 x 9.3 (6-7 x 8-10.5) microm, with an average length-width ratio of 1:1.4. A sporocyst residuum was present, located in between sporozoites, and was composed of numerous granules of unequal size. A small refractile body was present in each sporozoite. Collecting duct and distal renal tubular epithelial cells were distended by large oocysts in their cytoplasm, and many oocysts were present in the lumen of dilated tubules. Various stages of meronts, gamonts, and developing oocysts were present in other renal tubular epithelial cells. Multiple infections of parasitized cells were frequently observed, with cells containing up to 12 gamonts or developing oocysts. The importance of this Eimeria sp. on double crested cormorant populations is not known. But in this case, significant lesions and mortality were associated with infection. PMID- 12537119 TI - Qualitative evaluation of selective tests for detection of Neospora hughesi antibodies in serum and cerebrospinal fluid of experimentally infected horses. AB - Neospora hughesi is a newly recognized protozoan pathogen in horses that causes a myeloencephalitis similar to Sarcocystis neurona. There are no validated serologic tests using the gold standard sera that are currently available to detect specific N. hughesi antibodies and, thus, no tests available to detect antemortem exposure or estimate seroprevalence in the horse. The objectives of the present study were to establish a bank of gold standard equine sera through experimental infections with N. hughesi and to assess several serologic tests for the detection of related protozoan antibodies. Seven horses were inoculated with N. hughesi tachyzoites, and 7 horses received uninfected cell culture material. The horses were monitored, and blood and cerebrospinal fluid were collected repeatedly over a 4-mo period. With the sera, 4 different serologic techniques were evaluated. including a whole-parasite lysate enzyme-linked immunosorbent assay (ELISA), a recombinant protein ELISA, a modified direct agglutination test, and an indirect fluorescent antibody test. Qualitative and quantitative evaluation of the results showed that the N. hughesi indirect fluorescent antibody test (IFAT) consistently discriminated between experimentally infected and noninfected horses, using a cutoff of 1:640. Sera from 3 naturally infected horses had titers >1:640. Cerebrospinal fluid in all but I infected horse had very low N. hughesi IFAT titers (<1:160), starting at postinoculation day 30. PMID- 12537120 TI - Isolation of viable Toxoplasma gondii from naturally infected aborted bovine fetuses. AB - Neospora caninum and Toxoplasma gondii are related parasites. The former is a common cause of abortion in dairy cattle. The latter has not been conclusively demonstrated in bovine fetuses. During the course of attempts to isolate N. caninum from aborted fetuses, T. gondii was isolated from 2 aborted fetuses, 1 from Portugal and 1 from the United States. Both isolates were made by bioassay of fetal brains in mice. The fetus from Portugal was about 5 mo in gestational age, and the fetus from the United States was a full-term stillborn. PMID- 12537121 TI - The filarial endosymbiont Wolbachia sp. is absent from Setaria equina. AB - Wolbachia sp. was first reported in filarial nematodes over 25 yr ago. Today, much research is focused on the role of these bacteria in filarial worm biology. The filarial symbionts are closely related to arthropod symbionts, which are known to modify host reproduction and biology through various mechanisms. Similarly, it has been suggested that Wolbachia sp. is essential for long-term survival and reproduction of filariae. We report that Wolbachia sp. 16S rDNA was not found in the equine filarial nematode Setaria equina, using either polymerase chain reaction (PCR) or DNA hybridization. In addition, ultrastructural analysis of adult worms did not reveal the presence of Wolbachia sp. in hypodermal cords or reproductive tissues. These data suggest that like Onchocerca flexuosa and Acanthocheilonema vitae, S. equina may not be dependent on Wolbachia sp. for survival. PMID- 12537122 TI - Prevalence of antibodies to Neospora caninum and Sarcocystis neurona in sera of domestic cats from Brazil. AB - Parasite Biology, Epidemiology and Systematics Laboratory, Animal and Natural Resources Institute, Agricultural Research Service, United States Department of Agriculture, Building 1001, Beltsville, Maryland 20705-2350 Antibodies to Neospora caninum and Sarcocystis neurona were determined in serum samples of 502 domestic cats from Brazil using direct agglutination tests with the respective antigens. Antibodies to S. neurona were not found in 1:50 dilution of any serum in the S. neurona agglutination test. suggesting that domestic cats from Sao Paulo city were not exposed to S. neurona sporocysts from opossums. Antibodies to N. caninum were found in 60 (11.9%) of 502 cats with titers of 1:40 in 36 cats, 1:80 in 18 cats, 1:160 in 5 cats, and 1:800 in 1 cat using the Neospora agglutination test (NAT). Antibodies to N. caninum were confirmed by Western blotting in the sera of 10 cats with NAT titers of 1:80 to 1:800; this finding suggests that at least 10 cats had N. caninum-specific antibodies confirmed by 2 tests. This is the first documentation of natural exposure of cats to N. caninum. PMID- 12537123 TI - Effects of high temperature and disinfectants on the viability of Sarcocystis neurona sporocysts. AB - The effect of moist heat and several disinfectants on Sarcocystis neurona sporocysts was investigated. Sporocysts (4 million) were suspended in water and heated to 50, 55, 60, 65, and 70 C for various times and were then bioassayed in interferon gamma gene knockout (KO) mice. Sporocysts heated to 50 C for 60 min and 55 C for 5 min were infective to KO mice, whereas sporocysts heated to 55 C for 15 min and 60 C or more for 1 min were rendered noninfective to mice. Treatment with bleach (10, 20, and 100%), 2% chlorhexidine, 1% betadine, 5% o benzyl-p-chlorophenol, 12.56% phenol, 6% benzyl ammonium chloride, and 10% formalin was not effective in killing sporocysts. Treatment with undiluted ammonium hydroxide (29.5% ammonia) for 1 hr killed sporocysts, but treatment with a 10-fold dilution (2.95% ammonia) for 6 hr did not kill sporocysts. These data indicate that heat treatment is the most effective means of killing S. neurona sporocysts in the horse feed or in the environment. PMID- 12537124 TI - Giardia in beaver (Castor canadensis) and nutria (Myocastor coypus) from east Texas. AB - We examined the prevalence of Giardia sp. infection in nuisance beaver (Castor canadensis) and nutria (Myocastor coypus) in east Texas. From October 1992 through September 1993, 100 beaver and 30 nutria were collected in routine wildlife management activities conducted by the U.S. Department of Agriculture Animal and Plant Health Inspection Service-Texas Animal Damage Control Service. Fecal and duodenal mucoid samples were preserved from each animal. Fecal samples were examined for the occurrence of Giardia sp. cysts using the Merifluor immunoassay detection kit: 30 beaver (30.0%) and 20 nutria (66.7%) were positive for Giardia sp. Duodenal mucoid samples were examined for Giardia sp. trophozoites using trichrome staining. with 26 beaver (26.0%) and 20 nutria (73.3%) testing positive. Combining both techniques resulted in 33 beaver (33.0%) and 22 nutria (73.3%) testing positive for Giardia sp. We found no relationship between Giardia sp. and host age, sex, river system, habitat, county, or season in beaver. However, a relationship was found when season and habitat were considered together. No relationship was found between Giardia sp. and age, river system, habitat, county, or season in nutria: however, more males (87.5%) were infected than females (46.4%). PMID- 12537125 TI - A PCR-based RFLP analysis of Sarcocystis cruzi (Protozoa: Sarcocystidae) in Yunnan Province, PR China, reveals the water buffalo (Bubalus bubalis) as a natural intermediate host. AB - A polymerase chain reaction-based restriction fragment length polymorphism (RFLP) approach is used to examine Sarcocystis cruzi-like taxa from the atypical intermediate host, water buffalo, in Yunnan, People's Republic of China. The loci examined lie within the 18S rRNA gene. A total of 15 water buffalo isolates are compared with those of 10 S. cruzi from cattle. RFLP patterns for the S. cruzi isolates from cattle and the S. cruzi-like taxon from water buffalo are found to be identical with all the 12 restriction enzymes used. Interpopulation variation between samples from Kunming and Gengma (Yunnan) is found to be undetectable at these loci for both S. cruzi and the S. cruzi-like taxon. But RFLPs are found between the S. cruzi taxa and S. suihominis from pigs at the same study sites. These findings support the hypothesis that S. cruzi is able to use the water buffalo as an intermediate host and is not restricted to cattle as was previously supposed. PMID- 12537126 TI - Seroprevalence of antibodies to Neospora caninum in dogs from Spain. AB - The prevalence of Neospora caninum antibodies was determined in sera of 139 dogs from Catalonia (northeastern Spain) using the indirect immunofluorescence antibody test (IFAT). Antibodies in the IFAT were found in 17 of 139 dogs (12.2%) with titers ranging from 1:50 to 1: 1,600. Seroprevalence was higher in dogs over 1 yr old compared with dogs younger than 1 yr (P < 0.05). No statistical difference was observed when sex, breed, purpose, or modus vivendi was compared with seropositivity. Most dogs had low antibody titers, which indicated subclinical infection in the area studied. No neosporosis-related disease was reported from any dog, although a German shepherd with an antibody titer of 1:800 showed pododermatitis. All sera were also screened using a commercial direct agglutination test (DAT). The DAT showed a similar specificity but a lower sensitivity when compared with IFAT as a reference technique. PMID- 12537127 TI - Crab louse infestation in pre-Columbian America. AB - Until now, Pthirus pubis infestation in ancient human populations had only been recorded in the Old World. We found crab lice on South American mummified bodies from the Atacama Desert region. Crab louse eggs were found attached to the pubic hairs of a 2,000-yr-old Chilean mummy. Well-preserved adults were found in sediment and clothing from a Peruvian mummy dated 1,000 yr ago. Paleoparasitological evidence expands the knowledge of the distribution of this ectoparasite in ancient populations. As with many other parasites, pubic lice recorded in Andean populations show the antiquity of this parasite in the New World. It is likely that P. pubis entered the continent with early human migration to the New World. PMID- 12537128 TI - Relationship between helminth parasites and demographic attributes of a population of the subterranean rodent Ctenomys talarum (Rodentla: Octodontidae). AB - The helminth parasite fauna of a natural population of the octodontid, Ctenomys talarum, was studied. Parasites that were found included the nematodes Heligmostrongylus sp. and Trichuris sp. Total prevalence of parasitism was 92.3%, mean intensity of infection was 22.7 worms, and mean abundance was 21 worms. Prevalence and mean abundance of infection with Heligmostrongylus sp. were higher in C. talarum males relative to females. Ecological and physiological causes, as well as the mating system of the host species, influence the likelihood of sex differences in parasite infection. The low parasite burden and diversity of C. talarum are associated with restrictions imposed by the subterranean habitat and with life-history traits of these rodents. Whether these findings apply to other Ctenomys spp. is unknown. PMID- 12537129 TI - Stochasticity accelerates nematode egg development. AB - Day-degrees models of nematode development assume that temperature stochasticity has no effect on the development rate of infective stages as long as the mean temperature is held constant. This assumption was tested in this study. Unembryonated Heterakis gallinarum eggs were subjected to nocturnal and diurnal daily temperature cycles at 12 and 17 C. respectively, and embryonation was compared with eggs subjected to similar stochastic daily cycles, in which random normal variations in the temperature were added to the 2 temperatures. The prediction that there is no effect of stochasticity was refuted. Embryonation of eggs subjected to variable daily cycles occurred significantly earlier than that of eggs subjected to deterministic daily cycles, suggesting that stochastic variation in temperature accelerated embryonation even though mean temperatures were the same. These findings show that the development time of H. gallnarum eggs is decreased by stochastic variation in temperature, which may have important implications for the effects of climate change on parasite availability. PMID- 12537130 TI - Biological and molecular characterization of a raccoon isolate of Trypanosoma cruzi from South Carolina. AB - Biological and molecular characteristics of a raccoon isolate of Trypanosoma cruzi (R36) were compared with those of a known virulent strain (Brazil). Included in the characterization were growth rate in liver infusion tryptose medium, infectivity for murine fibroblasts, intracellular amastigote replication and trypomastigote release rates, polymerase chain reaction (PCR) profiling of the mini-exon gene, isoenzyme and random amplified polymorphic DNA (RAPD) profiles, and in vivo virulence for C3H/HeJ mice. Similar growth curves were noted for both strains; however, infectivity and rates of intracellular amastigote replication and trypomastigote release were significantly lower for the R36 isolate than for the Brazil strain. To determine virulence, C3H/ HeJ mice were exposed intraperitoneally to the R36 isolate. No parasite was observed in blood by direct examination or in tissues by histology; however, T. cruzi was detected by PCR in tissues (quadriceps and spleen) at 21 days postinfection. Analyses of the mini-exon gene, isoenzyme, and RAPD profiles indicate that R36 is in the T. cruzi II group and the Brazil strain is in the T. cruzi I group. Although infectivity and virulence of the raccoon isolate were lower than those for the Brazil strain, autochthonous infections in the United States have been reported, which suggests the need for further study of local T. cruzi isolates. PMID- 12537131 TI - Coinoculation of Borrelia spp. with tick salivary gland lysate enhances spirochete load in mice and is tick species-specific. AB - C3H/HeN mice were inoculated with 10(6) spirochetes, either Borrelia burgdorferi strain N40 or the Portuguese strain of B. lusitaniae, PotiB2. Mice receiving spirochetes coinoculated with salivary gland lysate (SGL) demonstrated significantly higher spirochete loads in target organs as measured by quantitative real-time polymerase chain reaction. This effect was tick dependent, in that Ixodes ricinus SGL specifically enhanced B. lusitaniae load, whereas I. scapularis SGL specifically increased B. burgdorferi N40 load, but did not significantly affect the dissemination of B. lusitaniae. Protein profile analysis indicated at least 5 major protein differences between I. scapularis and I. ricinus SGL, which can possibly account for this specific tick-spirochete interaction. PMID- 12537132 TI - Superannuation guarantee changes: what you need to know. PMID- 12537133 TI - Critical shortage of diagnostic (and research) pathologists in Australia. PMID- 12537134 TI - Three Australian veterinarians working closely with the restoration of animal health services within the century's first nation, Timor-Leste (East Timor). PMID- 12537135 TI - Seeking out animals to kill. PMID- 12537136 TI - Odd sheep were found walking on their knees and in obvious agony. PMID- 12537137 TI - Effect of declining owned-cat population on veterinary practices in Sydney. AB - OBJECTIVES: To investigate relationships between declining owned-cat populations and cat-related clinical work and practice activities. DESIGN: Sydney practitioners were interviewed, and practice records were examined, from small animal veterinary practices (n = 56) in Sydney. Using data that covered a 4 year period, the age distribution of cats was examined and associated trends in cat related clinical work were analysed. RESULTS: Important changes in the pet population attending veterinary practices were found, with 38.2% of respondents reporting a fall in cat desexings from 1996 to 2000. CONCLUSIONS: While economic impact could not be determined, the qualitative data demonstrated that the majority of practices in the survey were active in the promotion of cat ownership. The study provides a useful model for future investigations. To facilitate this process and to enhance principals' knowledge of their practice's population dynamics, we advocate a minimum standard of data collection that records week-by-week activities and the deaths of pets. PMID- 12537138 TI - The surgical management of a chronic inflammatory oropharyngeal lesion utilising magnetic resonance imaging for accurate localisation in a dog. PMID- 12537139 TI - Cryptococcosis in ferrets: a diverse spectrum of clinical disease. AB - Cryptococcosis was diagnosed in seven ferrets (five from Australia; two from western Canada) displaying a wide range of clinical signs. Two of the ferrets lived together. One (5-years-old) had cryptococcal rhinitis and presented when the infection spread to the nasal bridge. Its sibling developed cryptococcal abscessation of the right retropharyngeal lymph node 12 months later, soon after developing a severe skin condition. DNA fingerprinting and microsatellite analysis demonstrated that the two strains isolated from these siblings were indistinguishable. Two ferrets (2- to 3-years-old) developed generalised cryptococcosis: one had primary lower respiratory tract disease with pneumonia, pleurisy and mediastinal lymph node involvement, while in the other a segment of intestine was the primary focus of infection with subsequent spread to mesenteric lymph nodes, liver and lung. The remaining three ferrets (1.75 to 4-years-old) had localised disease of a distal limb, in one case with spread to the regional lymph node. Cryptococcus bacillisporus (formerly C. neoformans var gattii) accounted for three of the four infections in Australian ferrets where the biotype could be determined. The Australian ferret with intestinal involvement and the two ferrets from Vancouver had C. neoformans var grubii infections. PMID- 12537140 TI - Using modelling techniques to estimate epidemiological characteristics when information is scarce. PMID- 12537141 TI - The utility of Bayes' theorem and Bayesian inference in veterinary clinical practice and research. PMID- 12537142 TI - The estimated prevalence of Johne's disease infected sheep flocks in Australia. AB - OBJECTIVE: To estimate the likely geographical distribution and flock-prevalence of ovine Johne's disease (OJD) in Australia. DESIGN: A cross-sectional study design was used. PROCEDURE: The results of abattoir surveillance for OJD carried out during 2000 were analysed to estimate the prevalence of infected flocks in three regions of New South Wales and in other States. A Bayesian approach was used to adjust apparent prevalence estimates for the assumed flock-sensitivity and flock-specificity of abattoir surveillance, and to allow for uncertainty about the true values of these measures. RESULTS: The 95% probability limits for flock-prevalence at 31 December 2000 were 0.04%-1.5%, 8%-15% and 29%-39% for low, moderate and high prevalence regions of New South Wales respectively. The other States generally had an upper 97.5% probability limit of about 1% or less. Based on these estimates about 6 to 10% of flocks in New South Wales and 2.4 to 4.4% of flocks Australia-wide are likely to be infected. CONCLUSION: This study suggests that OJD has a highly clustered distribution in Australia, and provides estimates of the prevalence of infected flocks by State or region. Based on this analysis there were probably between 2000 and 3700 infected flocks in Australia at 31 December 2000, with more than 80% of these in a relatively small geographic area of central and southern New South Wales. Some States, such as Queensland and Western Australia, may have a prevalence equal or close to 0%, however the technique used was unable to demonstrate the absence of infection in these States with the intensity of surveillance undertaken to date. PMID- 12537143 TI - Herbert Robert Seddon Dvsc (1887-1964). PMID- 12537144 TI - Vitamin B12 injection for preventing cobalt deficiency in lambs. PMID- 12537145 TI - Health-education partnerships: a reflection on professional and personal past. PMID- 12537146 TI - Scholarly productivity: are nurse academics catching up? AB - The aim of this study was to document the amount of recent change in Australian nurse academics' scholarly productivity and to investigate the influence of demographic factors such as gender, academic rank, qualifications, increase in qualifications, state of residence, university and university size. Scholarly productivity was calculated from an audit of journal articles. The findings of this study indicate that, while there has been a slight increase in scholarly productivity in the last five years, nursing still lags behind other disciplines. Scholarly productivity was found to be positively associated with highest academic qualification, academic rank and promotion. The study indicates the continuing need for senior nurse academics to provide mentoring to colleagues and foster the development of skills associated with scholarly productivity. PMID- 12537147 TI - Non-adherence to HIV antiretroviral medications: 'The drugs are working and I'm a continuing success story'. AB - Non-adherence to antiretroviral (ARV) regimens has been associated with HIV drug resistance. Anecdotal evidence in our client group, people living with HIV/AIDS infection in Central Sydney Area Health Service (CSAHS), suggested that non adherence was an issue. Eighty three participants completed an anonymous questionnaire; with a 98% response rate. The mean age of respondents was 44 years with 68% over age 40 years. 68% of respondents missed doses of ARV, 54% because they forgot. 54% stated they did not report the non-adherence to their doctor. They were more likely to miss the lunch time dosage (n=49, p<0.05). The results demonstrate that non-adherence with medications is a significant issue and further research into strategies for interventions to aid adherence is required. PMID- 12537148 TI - Predictors of influenza vaccination amongst Australian nurses. AB - The Centers for Disease Control and Prevention recommend that health care workers receive influenza immunisation annually. There is no available data on factors predicting influenza immunisation or nurses' acceptance of the need for immunisation in Australia. To determine the predictors of influenza immunisation amongst nurses in Australia, a cross sectional survey of nurses in a 200-bed hospital was conducted. 232 of the 290 questionnaires were returned. In a logistic regression model, nurses having had previous influenza vaccination, severity of influenza if contracted, and, knowledge that the vaccine does not lead to clinical signs of influenza, were the factors most predictive of intention of influenza vaccination in the immediate future. Influenza immunisation rates among nurses may be increased by strategies addressing misconceptions about influenza immunisation and reinforcing predictors of influenza immunisation through education programs targeted to nurses. PMID- 12537149 TI - Systematic reviews of interpretive research: interpretive data synthesis of processed data. AB - This paper discusses the synthesis of qualitative data during the systematic review of interpretive research. Current approaches to data synthesis can be broadly classified as either descriptive or interpretive. Descriptive data synthesis is achieved through narrative and tabulation. While a number of different terms have been used, interpretive synthesis is achieved through processes similar to those used by primary interpretive researchers. However, there has been only limited professional discussion and debate of this issue and existing methods are in need of further investigation and the validity and usefulness of the product of this synthesis examined. PMID- 12537150 TI - Emotion and cardiac technology: an interpretive study. AB - This paper presents a frequently overlooked aspect of advanced technological care -that of the human dimension and emotions. Emotionality is defined as the emotional ways that a client experiences their embodied experience as a recipient of a cardiac pacemaker. One individual's story from a larger interpretive study of clients who received pacemakers is presented and interpreted. Kev's story encapsulates the difficulties of dealing with and understanding cardiac technology. When Kev's heart malfunctions he confronts a new reality; an experience where the 'technological body' is linked confusingly with emotion. This complex interplay between technology, the body and emotionality is discussed to demonstrate the importance of the mediating role that nurses can and should play in clients' adaptation and recovery. PMID- 12537151 TI - Living with breast cancer: the influence of rurality on women's suffering and resilience. a postmodern feminist inquiry. AB - This paper focuses on rural living as a dimension of women's experiences of living through breast cancer. The findings presented emanate from a feminist narrative research project that examined the experiences of rural women from south-west Queensland who were long-term survivors of breast cancer. This project aimed to listen, report and interpret rural women's stories of resilience in surviving breast cancer and moving on with their lives. The participants reported that there were both positive and negative aspects of living in a rural setting, especially when ill. Eight of the nine participants, however, felt strongly that the positive aspects of rural living outweighed the difficulties. This suggests that rurality in the context of health and illness must be considered as a multifaceted dimension, with resources to support cancer survivors building on the existing strengths in rural communities. PMID- 12537152 TI - The prevalence and correlates of consanguineous marriages in Yemen: similarities and contrasts with other Arab countries. AB - Using data on 9762 women from the 1997 Yemen Demographic and Maternal and Child Health Survey, this paper examines the prevalence and socioeconomic correlates of consanguineous marriages in Yemen. The results indicate that 40% of marriages are consanguineous, over 85% of which are between first cousins. The prevalence of consanguineous marriages appears to have increased over time, particularly for the last marriage cohort. As for socioeconomic correlates, the study confirms the inverse association between consanguineous marriages and women's education and occupation, age at marriage and economic status. However, no statistically significant difference in the prevalence of consanguinity has been found by place of residence and geographical region. Somewhat unexpected results have been obtained by husband's background characteristics, with higher educated men and those working in the modern sector of the economy being more likely to be married to cousins. PMID- 12537153 TI - The impact of rural-urban migration on under-two mortality in India. AB - This paper examines the impact of rural-urban migration on under-two mortality in India, using data from the 1992/93 Indian National Family Health Survey. Multilevel logistic models are fitted for mortality in three age groups: neonatal, early post-neonatal, and late post-neonatal and toddler. Migration status was not a significant determinant of mortality in any of the three age groups. Further analysis shows that a relationship between migration status and mortality exists when socioeconomic and health utilization variables are omitted from the models. The relationship between migration and mortality is thus explained by differences in socioeconomic status and use of health services between rural-urban migrant and nonmigrant groups. The selectivity of rural-urban migrants on socioeconomic characteristics creates mortality differentials between rural-urban migrants and rural non-migrants. Problems faced by migrants in assimilating into urban societies create mortality differentials between rural urban migrants and urban non-migrants. These results highlight the need to target migrants in the provision of health services, and demonstrate that rural areas continue to have the highest levels of infant-child mortality. Further research is needed to understand the health care needs of rural-urban migrants in order to inform the provision of appropriate health care. PMID- 12537154 TI - Secular gains in fluid intelligence: evidence from the Culture-Fair intelligence test. AB - There is no doubt about the reality of the secular increase in cognitive test scores. However, there is disagreement about a key issue: does the observed increase reflect a genuine upward trend in intelligence? Evidence from the Raven test is clear, although there are some doubts about its adequacy as a fine grained measure of fluid intelligence. Evidence from the so-called 'method of correlated vectors' is much less clear. When a crystallized battery is considered, the results leave little doubt: the increase does not reflect gains in general intelligence. However, when a fluid battery is analysed, the increase does reflect gains in general intelligence. The present study uses one of the best available measures of fluid intelligence (the Culture-Fair intelligence test) to provide new evidence for the secular increase in fluid intelligence, beyond the findings from the Raven test and the method of correlated vectors. A total of 4498 Spanish high school students and high school graduates were tested within a time interval of 20 and 23 years, respectively. The results show that there is a clear upward trend in intelligence. Moreover, students show an average increase equivalent to 6 IQ points, while graduates show an average increase of 4 IQ points. Therefore, more selected people (graduates) show a smaller increase than less selected people (students). Some implications are discussed. PMID- 12537155 TI - Pills, injections and audiotapes: reaching couples in Pakistan. AB - An innovative social marketing intervention in Pakistan distributes audiocassettes via chemist shops and Lady Health Visitors (LHVs) to reach women in a segregated society with accurate information on hormonal contraceptives. Operations research was done to assess the utility of the cassette in knowledge dissemination and adoption of hormonal use. In total 187 structured questionnaires were completed with couples who had obtained a cassette. Listeners were significantly more knowledgeable than non-listeners about correct use of hormonals (OR = 8.6 for women and OR = 12.7 for men). Hormonal use increased from 12% to 25%. LHVs also organized discussion groups for women, and attending such a chat group was the strongest predictor for adoption of pills and injectables (OR = 4.15). Equivalent male groups are suggested to reach apprehensive men. By providing accurate information to urban couples and by acquiring a knowledgeable critical mass of satisfied users, the cassette could be a powerful catalyst to further contraceptive diffusion. PMID- 12537156 TI - Determinants of waiting time to third pregnancy using censored linear regression. AB - The intervals between pregnancies have important effects on fertility and maternal and infant health outcomes. This study uses linear regression with censored observation to assess the determinants of the waiting time to third pregnancy. The analysis is applied to data from the Second Malaysian Family Life Survey consisting of 1172 women who had their second delivery ending in a live birth. Contraceptive use, age of the woman, duration of breast-feeding, length of previous pregnancy interval and education of the woman all affect the waiting time to third pregnancy significantly. PMID- 12537157 TI - Adolescents' decision-making and attitudes towards abortion in north-east Brazil. AB - Three groups of adolescents are compared with regard to their own considerations of abortion and when they believe abortion is justified. One group of adolescents terminated their pregnancies (n=95), a second became pregnant and carried their pregnancies to term but considered abortion (n=68), and the third also carried their pregnancies to term but did not consider abortion (n=204). The study was carried out between 1995 and 1998 in Fortaleza, Brazil. Adolescents were interviewed at the time of their hospitalization or their first prenatal visit and again at 6 weeks and 1 year post-abortion or postpartum. Friends and family recommended abortion to at least half of the teenagers in each group. Teenagers who aborted were more accepting of abortion than those who did not abort, while those who considered abortion found the practice more justified than those who did not consider abortion. Teenagers who aborted became less accepting a year later, while those who did not consider abortion became more accepting. A better understanding of adolescent attitudes towards abortion and their decision-making process should help adults and professionals meet the needs of adolescents for support in the process and in the reduction of the number of unintended pregnancies in the future. PMID- 12537158 TI - Inbreeding pattern and reproductive success in a rural community from Galicia (Spain). AB - The present paper is based on information on marital patterns and reproductive performance (reconstituted families) in the rural community of Los Nogales, Galicia, Spain. Biodemographic data were used to analyse temporal changes and structure of consanguinity in the population, as well as consanguinity versus reproductive success. Of 2347 marriages celebrated between 1871 and 1977, 5.15% were between relatives (up to third degree). The population's inbreeding coefficient was above the Spanish average after 1890, and levels were high during the first part of the 20th century. A possible underestimation of inbreeding coefficients is attributed to the elevated frequency of extramarital births occurring in the Los Nogales population (13.47%). The structures of consanguineous matings, expressed as C22/C33 and C33/C44, turned out to be 0.86 and 0.82 respectively. Regarding migration, before 1920 consanguinity increased, but net migration was negative. In the period 1920-1929 consanguinity began to decrease, prior to maximum emigration. As for reproductive success, information from 1503 reconstituted families shows that complete fertility was slightly higher among consanguineous families despite a greater infant mortality. Lower infertility in consanguineous matings, as well as lower age at marriage and first maternity, could explain the above results. PMID- 12537159 TI - Seasonality of births is associated with seasonality of marriages in Malta. AB - This study was carried out to quantify secular trends in seasonal variation in births in Malta, a small Mediterranean country where the vast proportion of births occur in wedlock due to a predominantly Roman Catholic population. It also related such variations to seasonal variation in marriages. Annual seasonal peaks of marriages and births were analysed over the period 1950-1996 by X11 ARIMA. A significant peak in marriages (n = 111,932) in the third quarter of the year was found for almost the entire period under study. This was paralleled by a peak in births (n = 299,558) for the period 1970-1996, which lagged after the peak in marriages by 13-14 months. For the period 1994-1996, when monthly data for monthly pregnancies were available by pregnancy order, the peak in births was caused by first pregnancies only. Seasonal patterns in births occur almost universally due to cultural and/or biometeorological factors. The best known patterns include those of the southern United States, where births decline in April and May, and in northern Europe, where births peak in March and April. In Malta, the late summer peak in births appears to be due to a practical and planned approach by Maltese couples to contraceptive planning, probably influenced by the Roman Catholic ethos and social pressures, with unprotected intercourse occurring only after marriage. In Malta, birth control, albeit by so called natural methods, was introduced in the 1960s. Prior to this period, births peaked towards the beginning/end of the year, and this may be the more natural seasonality of births in Malta. PMID- 12537160 TI - The effect of obesity on overall, circulatory disease- and diabetes-specific mortality. AB - This paper explores the relationship between body mass and risk of death among US adults. The National Health Interview Survey-Multiple Cause of Death linked data set is used for the years 1987-1997, and Cox proportional hazard models are employed to estimate the association between obesity, as measured by the body mass index (BMI), and overall, circulatory disease-specific and diabetes-specific mortality. A U-shaped relationship is found between BMI and overall mortality. Compared with normal weight individuals, mortality during the follow-up period is 34% higher among obese class II individuals and 77% higher among obese class III individuals, controlling for age and sex. A J-shaped relationship exists between circulatory disease mortality and obesity, with a slightly higher risk of death for all categories of BMI. The relationship between BMI and diabetes mortality is striking. Compared with normal weight individuals, obese class I individuals are 2.8 times as likely to die, obese class II individuals are 4.7 times as likely to die, and obese class III individuals are 9.0 times as likely to die of diabetes during the follow-up period, controlling for age and sex. These results demonstrate that obesity heightens the risk of overall and circulatory disease mortality, and even more substantially increases the risk of diabetes mortality. These mortality findings, together with the substantial recent increases in obesity, lend urgency to public health programmes aimed at reducing the prevalence and consequences of obesity. PMID- 12537161 TI - Role of health-seeking behaviour in child mortality in the slums of Karachi, Pakistan. AB - The role of family health-seeking behaviour in under-five-year child mortality was explored through the combined approach of examining health-seeking behaviour regarding treatment generally, and in specific in relation to illness before death. A population-based case control study was carried out during the period 1993-1994 using 222 deaths from diarrhoea and acute respiratory illness (ARI) in children under five years of age in six slums of Karachi as cases, and 419 controls matched on age, disease (diarrhoea and ARI) and slum. Factors significantly associated (p<0.05) with child mortality in the multivariate analysis were: mothers changing healers quickly, using a traditional healer or an unqualified doctor and mothers to whom doctors did not explain the treatment, even when maternal education was controlled for. Seeking effective medical services is highly influential on whether the child survives or succumbs to ARI or diarrhoea. As mothers are the first providers of care, an attempt should be made to try and improve their skills through health education so that they can use simple and effective treatments for minor illnesses. They should also be taught to recognize potentially life-threatening conditions, to seek care early and to persist with treatment. PMID- 12537162 TI - Sex differences on the progressive matrices: some data from Hong Kong. AB - There has been a debate between N. J. Mackintosh and the first author of this paper on whether the sex difference on the Progressive Matrices is zero or, at most, 1-2 IQ points in favour of either sex, as maintained by Mackintosh, or whether from the age of 15 years onwards males obtain higher average scores than females by more than 2 IQ points, as maintained by Lynn. New data relevant to this controversy are presented from Hong Kong consisting of sex differences on the standardization sample of the Advanced Progressive Matrices on 15- to 18-year olds. The results are that males obtained a higher mean score than females of 1.6 raw score points, equivalent to an advantage of 3.2 or 4.1 IQ points, according to two alternative methods of calculation. The results provide further confirmation that in later adolescence and among adults, males obtain significantly higher mean IQs on the Progressive Matrices than females. PMID- 12537163 TI - Family physicians increase provision of well-infant care despite decline in prenatal services. AB - Over the past 20 years, family physicians and general practitioners (FP/GPs) and pediatricians have upheld their commitment to preventive care for infants. Non Metropolitan Statistical Areas (non-MSAs) depend on family physicians for almost one half of their well-infant care. FP/GPs have increased their overall provision of well-infant care despite a decline in delivery of prenatal services. This commitment to child health care demands continued excellence of family physician training in pediatric medicine, preventive care, and child advocacy. PMID- 12537164 TI - Why is aspirin a contraindication for trigger-point injections? PMID- 12537165 TI - How effective is bracing for treatment of scoliosis? PMID- 12537166 TI - Complementary and alternative medicine: examining the evidence. PMID- 12537167 TI - Are metered-dose inhalers with holding chambers better than nebulizers for treating acute asthma? PMID- 12537168 TI - Diagnosing secondary hypertension. AB - Secondary hypertension is elevated blood pressure that results from an underlying, identifiable, often correctable cause. Only about 5 to 10 percent of hypertension cases are thought to result from secondary causes. The ABCDE mnemonic can be used to help determine a secondary cause of hypertension: Accuracy of diagnosis, obstructive sleep Apnea, Aldosteronism, presence of renal artery Bruits (suggesting renal artery stenosis), renal parenchymal disease (Bad kidneys), excess Catecholamines, Coarctation of the aorta, Cushing's syndrome, Drugs, Diet, excess Erythropoietin, and Endocrine disorders. An algorithm showing the general strategy to help screen for factors involved in secondary hypertension is presented. Routine urinalysis, complete blood cell count, blood chemistry profile (potassium, sodium, creatinine, fasting glucose, fasting lipid levels), and a 12-lead electrocardiogram are recommended for all patients with hypertension. PMID- 12537170 TI - Information from your family doctor. Echinacea: what should I know about it? PMID- 12537169 TI - Echinacea. AB - Echinacea is the name of a genus of native North American plants, commonly known as the purple coneflower. The most widely used herbal product in the United States is a liquid extract made from the root of Echinacea purpurea. Because the active component of the plant has not been identified, commercial echinacea products are not typically standardized to any particular component. The research literature on echinacea is difficult to evaluate because of the heterogeneity of the products used in various studies. The herb has been recommended as a prophylactic treatment for upper respiratory infection and is widely used for this indication. However, based on the current literature, it appears that prophylactic echinacea does not have a significant impact on the frequency, severity, or duration of upper respiratory infection. The data regarding treatment of upper respiratory infection appear to support a modest positive effect. No significant herb-drug interactions with echinacea have been reported; adverse effects reported generally have been uncommon and minor, including abdominal upset, nausea, and dizziness. PMID- 12537171 TI - Tarsal navicular stress fractures. AB - Stress fractures of the tarsal navicular bone are being recognized with increasing frequency in physically active persons. Diagnosis is commonly delayed, and outcome often suffers because physicians lack familiarity with the condition. Navicular stress fractures typically present in a running athlete who has gradually increasing pain in the dorsal mid-foot with occasional radiation of pain down the medial arch. Because initial plain films are often normal, the next diagnostic test of choice is triple-phase bone scan, which is positive early in the process and localizes the lesion well. After a positive bone scan, a computed tomographic scan should be obtained to provide anatomic detail and guide therapy. Nondisplaced, noncomminuted fractures respond well to six weeks of non-weight bearing cast immobilization. Displacement, comminution, and delayed or nonunion fractures are indications for surgical open reduction internal fixation. PMID- 12537172 TI - Perioperative management of diabetes. AB - Maintaining glycemic and metabolic control is difficult in diabetic patients who are undergoing surgery. The preoperative evaluation of all patients with diabetes should include careful screening for asymptomatic cardiac or renal disease. Frequent self-monitoring of glucose levels is important in the week before surgery so that insulin regimens can be adjusted as needed. Oral agents and long acting insulin are usually discontinued before surgery, although the newer long acting insulin analog glargine may be appropriately administered for basal insulin coverage throughout the surgical period. The usual regimen of sliding scale subcutaneous insulin for perioperative glycemic control may be a less preferable method because it can have unreliable absorption and lead to erratic blood glucose levels. Intravenous insulin infusion offers advantages because of the more predictable absorption rates and ability to rapidly titrate insulin delivery up or down to maintain proper glycemic control. Insulin is typically infused at 1 to 2 U per hour and adjusted according to the results of frequent blood glucose checks. A separate infusion of dextrose prevents hypoglycemia. Potassium is usually added to the dextrose infusion at 10 to 20 mEq per L in patients with normal renal function and normal preoperative serum potassium levels. Frequent monitoring of electrolytes and acid-base status is important during the perioperative period, especially in patients with type 1 diabetes because ketoacidosis can develop at modest levels of hyperglycemia. PMID- 12537173 TI - Dermatophyte infections. AB - Dermatophytes are fungi that require keratin for growth. These fungi can cause superficial infections of the skin, hair, and nails. Dermatophytes are spread by direct contact from other people (anthropophilic organisms), animals (zoophilic organisms), and soil (geophilic organisms), as well as indirectly from fomites. Dermatophyte infections can be readily diagnosed based on the history, physical examination, and potassium hydroxide (KOH) microscopy. Diagnosis occasionally requires Wood's lamp examination and fungal culture or histologic examination. Topical therapy is used for most dermatophyte infections. Cure rates are higher and treatment courses are shorter with topical fungicidal allylamines than with fungistatic azoles. Oral therapy is preferred for tinea capitis, tinea barbae, and onychomycosis. Orally administered griseofulvin remains the standard treatment for tinea capitis. Topical treatment of onychomycosis with ciclopirox nail lacquer has a low cure rate. For onychomycosis, "pulse" oral therapy with the newer imidazoles (itraconazole or fluconazole) or allylamines (terbinafine) is considerably less expensive than continuous treatment but has a somewhat lower mycologic cure rate. The diagnosis of onychomycosis should be confirmed by KOH microscopy, culture, or histologic examination before therapy is initiated, because of the expense, duration, and potential adverse effects of treatment. PMID- 12537174 TI - An office-based approach to influenza: clinical diagnosis and laboratory testing. AB - Vaccination is the primary measure for preventing morbidity and mortality from influenza. During the influenza season, family physicians must distinguish influenza from the common cold and other flu-like illnesses. Signs and symptoms of influenza include abrupt onset of fever, severe myalgias, anorexia, sore throat, headache, cough, and malaise. Clinical diagnosis can be difficult or nonspecific when patients have other symptoms (e.g., stuffy nose, sneezing, cough, sore throat) that can be caused by various respiratory viruses or bacterial pathogens. Family physicians can improve diagnostic accuracy by being aware of the epidemiology of influenza. During outbreaks of influenza, commercially available rapid assays can be used to identify type A and B viruses. On average, rapid in-office tests are more than 70 percent sensitive and 90 percent specific for viral antigens. The assays vary in complexity, specificity, sensitivity, time to obtain results, specimen analyzed, and cost. The results of rapid viral tests can guide treatment decisions. PMID- 12537175 TI - Colorectal cancer screening. PMID- 12537176 TI - Generalized anxiety disorder. PMID- 12537177 TI - Photo quiz. Skin rash in a patient with diabetes. PMID- 12537178 TI - Information from your family doctor. Choosing the right diet to lose weight. PMID- 12537180 TI - Information from your family doctor. Healthy food choices for losing weight. PMID- 12537179 TI - Information from your family doctor. What it takes to lose weight. PMID- 12537181 TI - Recommended childhood and adolescent immunization schedule, United States, 2003 and update on childhood immunizations. PMID- 12537182 TI - Negotiating a request for nondisclosure. PMID- 12537183 TI - Final thoughts on a year of significant change. PMID- 12537184 TI - New inside and out. PMID- 12537185 TI - Pork or beef? PMID- 12537186 TI - Comparison of bovine and porcine heparin in heparin antibody formation after cardiac surgery. AB - BACKGROUND: Heparin-induced thrombocytopenia (HIT) is a potentially devastating complication of heparin therapy. The incidence of clinical HIT after cardiovascular surgery is less than 2%, although asymptomatic antibodies to heparin-platelet factor 4 (PF4) occur more frequently. Bovine heparin is thought to cause more HIT than porcine heparin, although this has never been established for heparin use during coronary artery bypass grafting. We therefore undertook a randomized, prospective study of heparin-PF4 antibody formation in patients undergoing first-time CABG given intraoperative bovine or porcine heparin. METHODS: Two hundred seven patients (108 porcine, 99 bovine) completed the study. Heparin given pre- or postoperatively was always porcine. Platelet counts and heparin-PF4 antibody tests (enzyme-linked immunosorbent assays) were performed preoperatively and daily until postoperative day 7 or discharge if earlier. RESULTS: The overall incidence of heparin-PF4 antibody formation was 42%. Six patients (2.9%) were positive preoperatively, of which, 1 developed clinical HIT. When these were excluded, seroconversion rates were 44 of 99 (44.4%) and 33 of 108 (30.6%) for bovine and porcine heparin, respectively (p = 0.041). Among patients who produced antibodies, most (90% bovine, 85% porcine) seroconverted after postoperative day 2. There were no differences in postoperative platelet counts; only 1 patient developed thrombosis associated with seroconversion, but without developing thrombocytopenia. The seroconversion rates for patients having cardiopulmonary bypass or off-pump surgery were not significantly different. CONCLUSIONS: This study confirms the high frequency of heparin-PF4 antibodies after coronary artery bypass grafting and demonstrates a significantly higher incidence after bovine heparin. However, because some patients may seroconvert after discharge, our study may underestimate the true incidence. PMID- 12537187 TI - Retrograde autologous priming: is it useful in elective on-pump coronary artery bypass surgery? AB - BACKGROUND: The effect of reduced cardiopulmonary bypass (CPB) prime volume by retrograde autologous priming (RAP) was studied. METHODS: Twenty patients undergoing elective coronary artery bypass grafting were randomized to either standard prime (SP) volume (1,602 +/- 202 mL crystalloid prime, n = 10) or RAP (395 +/- 150 mL). RAP was performed by draining crystalloid prime from the arterial and venous lines into a recirculation bag before CPB. Cardiac index, pulmonary vascular resistance index, systemic vascular resistance index, alveolar arterial oxygen tension difference, pulmonary shunt fraction, extravascular lung water (EVLW), plasma colloid osmotic pressure (COP), crystalloid fluid balance, body weight, and clinical parameters were evaluated perioperatively. RESULTS: Demographic data and operative parameters were equal for patients in both groups. During CPB, COP was reduced by 55% in the SP group (9.8 +/- 2.0 vs 21.4 +/- 2.1 mm Hg) and by 41% in the RAP group (12.4 +/- 1.1 vs 20.9 +/- 1.8 mm Hg) (p = 0.008, SP vs RAP group). Compared with preoperatively, EVLW was unchanged in the RAP group 2 hours post-CPB, but it was elevated by 21% in the SP group (p = 0.002, SP vs RAP group). End-CPB crystalloid fluid balance was significantly reduced in the RAP group (1,857 +/- 521 vs 2,831 +/- 637 mL). Postoperative (day 2) weight gain in the SP group (1.5 +/- 1.2 kg, p = 0.021) was absent in the RAP group (0.1 +/- 0.9, NS). Postoperative time to full mobilization was shorter in the RAP group. Postpump cardio-respiratory function did not differ among groups. CONCLUSIONS: This small-scale pilot study indicates that by reducing crystalloid fluid administration and fall of COP during CPB, RAP reduces postpump EVLW accumulation and weight gain in uncomplicated coronary artery bypass graft patients with no associated effects on cardio-respiratory function. PMID- 12537188 TI - Durability and outcome of aortic valve replacement with mitral valve repair versus double valve replacement. AB - BACKGROUND: The purpose of this study was to evaluate morbidity and mortality after double valve replacement (DVR) and aortic valve replacement with mitral valve repair (AVR + MVP). METHODS: From 1977 to 2000, 379 patients underwent DVR (n = 299) or AVR + MVP (n = 80). Actuarial survival and freedom from reoperation were determined by the Kaplan-Meier method. Potential predictors of mortality and reoperation were entered into a Cox multiple regression model. Propensity score was introduced for the multivariable regression modeling for adjustment of a selection bias. RESULTS: Survival 15 years after surgery was similar between the groups (DVR, 81% +/- 3%; AVR + MVP, 79% +/- 7%; p = 0.44). Freedom from thromboembolic event at 15 years was similar between the groups (p = 0.25). Freedom from mitral valve reoperation at 15 years was significantly better for the DVR group (54% +/- 5%) as compared with the AVR + MVP group (15% +/- 6%; p = 0.0006), primarily due to progression of mitral valve pathology and early structural deterioration of bioprosthetic aortic valve used for patients with AVR + MVP. After AVR + MVP, freedom from mitral reoperation at 15 years was 63% +/- 16% for nonrheumatic heart diseases, and 5% +/- 5% for rheumatic disease (p = 0.04). CONCLUSIONS: Although both DVR and AVR + MVP provided excellent survival, DVR with mechanical valves should be the procedure of choice for the majority of patients because of lower incidence of valve failure and similar rate of thromboembolic complications compared with AVR + MVP. MVP should not be performed in patients with rheumatic disease because of higher incidence of late failure. PMID- 12537189 TI - Clinical and hemodynamic evaluation of small Perimount aortic valves in patients aged 75 years or older. AB - BACKGROUND: There is the potential for iatrogenic aortic stenosis and poor quality of life when small aortic valve bioprostheses are used in elderly patients. The alternative is enlarging the aortic annulus to accommodate larger size prostheses, increasing operative mortality. It was hoped that bovine pericardial valves would improve hemodynamic performance in the smaller valve sizes. METHODS: To determine long-term results and in vivo hemodynamic performance of small-size aortic Carpentier-Edwards bovine pericardial valves (Perimount) in elderly patients, we analyzed our follow-up and echocardiographic data from patients 75 years of age or older receiving isolated 19-mm and 21-mm Perimount valves. Ninety-four patients with a mean age of 77 +/- 2.2 years were followed for 12 years. Seventeen patients with 19-mm and 25 patients with 21-mm Perimount valves underwent transthoracic echocardiograms. RESULTS: Operative mortality was 6.3% (6 of 94). Twelve-year survival was 82.7%. Freedom from thromboembolism was 86.9% at 12 years. Two patients had anticoagulation-related bleeding. Overall New York Heart Association class decreased from 3 +/- 1 to 1.6 +/- 0.7 at the end of follow-up. Hemodynamic performances were satisfactory in both 19-mm and 21-mm Perimount valves, with low peak and mean transvalvular gradients and good effective orifice areas, orifice area indices, and performance indices. CONCLUSIONS: Perimount aortic valve in the small aortic annulus has yielded excellent long-term results and hemodynamic performances. Perimount is a very satisfactory option in elderly patients. Implantation of a Perimount bioprosthesis avoids enlargement of the small aortic annulus, reducing mortality and morbidity associated with this procedure. PMID- 12537190 TI - Open mitral commissurotomy in the current era: indications, technique, and results. AB - BACKGROUND: The present retrospective study is focused on indications, techniques, and results of open mitral commisurotomy in the current era. METHODS: Of the 1,280 patients undergoing open-heart surgical procedures for rheumatic mitral stenosis between January 1990 and July 2000, 276 (21.6%) patients underwent open mitral commissurotomy. Major indications included presence of left atrial thrombus/clot (n = 82, 29.7%), severe subvalvular disease (n = 110, 39.8%), mitral valve calcification (n = 42, 15.2%), mild mitral regurgitation (n = 28, 10.0%), associated aortic valve disease (n = 55, 19.9%), organic tricuspid valve disease (n = 20, 7.2%), and failure or restenosis after closed or balloon mitral valvuloplasty (n = 55, 19.9%). Age of patients ranged from 7 to 67 years (mean, 30.2 +/- 12 years). The majority (76%) were in New York Heart Association class III or IV, and 6.9% were in congestive heart failure. Atrial fibrillation was present in 134 (48.6%) patients. Mitral valve area ranged from 0.3 to 0.7 cm2 (mean, 0.52 +/- 0.12 cm2). Mid-diastolic gradients across the mitral valve ranged from 8 to 34 mm Hg (mean, 14.5 +/- 6.2 mm Hg), and end-diastolic gradients ranged from 8 to 42 mm Hg (mean, 15.2 +/- 5.7 mm Hg). Open mitral commissurotomy was performed using standard cardiopulmonary bypass. Associated aortic valve procedure was performed in 55 patients, and either tricuspid valvotomy or repair was performed in 28 patients. RESULTS: There were four early deaths. All these patients had associated aortic valve procedure (Ross procedure in 2 and homograft aortic valve replacement in 2). Three patients developed severe mitral regurgitation in early postoperative period (< or = 30 days) and required reoperation. Predischarge echocardiography showed mitral valve area from 1.4 to 3.5 cm2 (mean, 2.6 +/- 0.6cm2) and moderate mitral regurgitation in 4 patients. Follow-up ranged from 1 to 130 months (mean, 64.5 +/- 28.6 months). There was no late death. There were three reoperations for mitral valve failure, and an additional 2 patients developed severe mitral stenosis (mitral valve area < 1.0 cm2). In operative survivors, freedom from mitral valve failure at 10 years was 87.0% +/- 3.5%. In patients with isolated open mitral commissurotomy, the incidence of thromboembolism was 0.5%/patient-year. CONCLUSIONS: Open mitral commissurotomy provides excellent early and long-term results in a selected group of patients. PMID- 12537191 TI - Tricuspid valve replacement: an analysis of 25 years of experience at a single center. AB - BACKGROUND: Tricuspid valve replacement is seldom used in clinical practice, but the choice between mechanical and biologic prostheses remains controversial. METHODS: Between 1977 and 2002, 97 patients underwent tricuspid valve replacement and were followed at the Montreal Heart Institute Valve Clinic. Patients underwent replacement with bioprostheses (n = 82) and mechanical valves (n = 15). RESULTS: Patients with bioprosthetic tricuspid replacements averaged 53 +/- 13 years of age compared with 48 +/- 11 years in those with tricuspid mechanical valve replacements (p = 0.2). Isolated tricuspid valve replacement was performed in 11 patients (73%) in the mechanical valve group compared with 31 patients (38%. p = 0.01) in the bioprosthetic replacement group. In patients undergoing bioprosthetic tricuspid replacement, 51 (62%) underwent multiple associated valve replacements. The 5-year survival after tricuspid replacement averaged 60% +/- 13% in the mechanical valve group and 56% +/- 6% in the biologic replacement group (p = 0.8). The 5-year freedom rate from tricuspid valve reoperation averaged 91% +/- 9% in patients with mechanical valves and 97% +/- 3% in those with biologic valves (p = 0.2). CONCLUSIONS; Patient survival after tricuspid valve replacement is suboptimal but related to the clinical condition at operation. The use of biologic prostheses for tricuspid valve replacement remains a good option in young patients because of limited life expectancy unrelated to the type of tricuspid prostheses at long-term follow-up. PMID- 12537192 TI - The effect of maze operations on atrial volume. AB - BACKGROUND: Unmodified maze III operations show long-term eradication of atrial fibrillation (AF) in more than 85% of patients with or without structural heart disease. The effect of this procedure on atrial volumes is not known. METHODS: Two patient populations were studied: (1) patients undergoing unmodified maze III operations combined with surgical structural heart disease, mostly mitral valve operations (group A; n = 32); and (2) patients with only AF selected for unmodified maze III operations (group B; n = 32). In groups A and B, transthoracic Doppler echocardiographic studies were prospectively made preoperatively, and at 3 and 12 months postoperatively. Left and right atrial dimensions and volumes and atrial contractions were determined and compared with base line patient characteristics and 12 months arrhythmia outcomes. RESULTS: One year postoperatively all patients were alive. In groups A and B, 92% were free of AF and other atrial arrhythmias. A significant reduction of left atrial volume at 1 year postoperatively was apparent in group A, whereas the left atrial volume did not change significantly in group B. The reduction observed in group A was not related to postoperative age, type or duration of AF, or late atrial arrhythmia outcome. In both groups the right atrial volume remained unchanged at 12 months postoperatively. CONCLUSIONS: The unmodified maze III operation does not affect atrial volume in patients without structural heart disease. In patients with structural heart disease, the mitral valve operation contributes to the reduction of left atrial volume and dimension by improving the hemodynamic condition. PMID- 12537193 TI - Surgical outcome of the maze procedure for atrial fibrillation in mitral valve disease: rheumatic versus degenerative. AB - BACKGROUND: The results of the maze procedure are known to be less satisfactory in rheumatic mitral disease than in nonrheumatic mitral valve disease. The aim of this study was to determine whether the etiology of mitral valve disease affected surgical outcome. METHODS: From July 1997 to January 2001, 129 consecutive patients with chronic atrial fibrillation associated with mitral valve disease had mitral valve operations with the maze procedure. The underlying mitral pathology was rheumatic in 86 patients (group R) and degenerative in 43 (group D). Echocardiograms and electrocardiograms were performed immediately and then repeated 3 months and 6 months postoperatively. RESULTS: The mean age, duration of atrial fibrillation, and preoperative left atrial size were similar between the groups. There was no operative mortality and no significant difference in cardiopulmonary bypass and aortic cross-clamp times. The sinus conversion rate at 7 days postoperatively was 86% in both groups, and at 6 months it was 95.3% in group R and 97.7% in group D (p > 0.05). The transmitral A wave detection rates in groups R and D at 7 days and 6 months postoperatively were, respectively, 63.1% versus 67.4% and 90.4% versus 91.9% (p > 0.05). The transmitral A wave velocity (cm/second) at the same times (7 days and 6 months postoperatively) was 41.9 +/- 41.6 versus 45.5 +/- 37.7 and 67.8 +/- 38.2 versus 69.8 +/- 35.8 in groups R and D, respectively (p > 0.05). CONCLUSIONS: The maze procedure is equally effective in treating chronic atrial fibrillation in patients with either rheumatic or nonrheumatic mitral valve disease in terms of sinus conversion rate and left atrial transport function. PMID- 12537194 TI - On-pump coronary artery surgery versus off-pump exclusive arterial coronary grafting: a matched cohort comparison. AB - BACKGROUND: It is unknown whether coronary artery bypass grafting without cardiopulmonary bypass and with exclusive use of arterial grafts (arterial off pump CABG) offers any significant short-term advantages over standard CABG with cardiopulmonary bypass. Accordingly, we performed a comparison of the short-term outcomes of arterial off-pump and standard CABG patients matched for preoperative risk and number of grafts. METHODS: We studied 90 consecutive arterial off-pump CABG patients during a 2-year period, obtained demographic and clinical features and surgical characteristics, and calculated their predicted surgical risk (EuroSCORE). Using a database of 750 contemporaneous patients treated with standard CABG, we created a matched cohort of 90 patients using an iterative process prioritizing number of grafts, target vessels, EuroSCORE, age, and sex. We compared the two groups for baseline features and short-term clinical outcomes. RESULTS: There were no differences in age (65.9 versus 64.7 years), sex, EuroSCORE (3.3 versus 3. 6), number of grafts (2.1 versus 2.1), and preoperative left ventricular function. Arterial off-pump CABG, however, was associated with decreased duration of operation (213 versus 252 minutes; p < 0.0013), decreased peak postoperative troponin I levels (mean, 10.8 versus 29.1 ng/mL; p < 0.0001), decreased peak norepinephrine dose (2.3 versus 4.1 microg/ min; p < 0.0082), and decreased likelihood of receiving red blood cell transfusion (17.8% versus 40%; p = 0.0016). There were no differences in duration of intensive care unit or hospital stay, incidence of atrial fibrillation, or other clinical complications. There was one death in each group. CONCLUSIONS: After matching for number of grafts and other important preoperative risk markers, arterial off-pump CABG still decreases the need for red blood cell transfusion and offers other moderate clinical advantages compared with standard on-pump CABG. PMID- 12537195 TI - Readmission and mortality in patients discharged the day after off-pump coronary bypass surgery. AB - BACKGROUND: The fate of patients discharged the day following off-pump coronary bypass (OPCAB) has not previously been reported. We studied the mortality and readmissions of a consecutive series of patients discharged after OPCAB, and compared the outcomes of those discharged the day following surgery to the rest of the group. METHODS: All patients having OPCAB through median sternotomy during the calendar year 2000 by a single surgeon were retrospectively reviewed. Demograghics, intraoperative variables and postoperative complications, readmissions and mortality were recorded. Factors were analyzed to determine associations with time of discharge and readmission. RESULTS: One hundred fifteen patients had isolated OPCAB averaging 3.1 grafts. Two patients (1.8%) died before discharge. Sixty-three of 113 patients (55.8%) were discharged on day 1 and 8 (12.7%) required readmission compared to 13 of 50 (26%) discharged later. Diabetes (p = 0.04) and renal failure (p = 0.01) exhibited univariate association with day 1 discharge while multivariate analysis added infarction. The combination of previous bypass, obesity, acute myocardial infarction, and hypertension was associated with readmission in the entire OPCAB group but not in day 1 discharged patients. CONCLUSIONS: The readmission rate for the entire group (18.6%) was high but lower in day 1 discharge patients (12.7%). Day 1 discharge (55.8%) was unusual in patients with diabetes, renal failure, or recent infarction. Previous bypass, obesity, acute myocardial infarction, and hypertension were associated with readmission for the entire group only. Day 1 discharged patients had no deaths or serious consequences, and there were no readmissions in more than 87%. PMID- 12537196 TI - Variation in mortality risk factors with time after coronary artery bypass graft operation. AB - BACKGROUND: Differences in mortality risk factor sets during different time periods (eg, short-term versus intermediate-term) after coronary artery bypass grafting have been reported. However, little is known about the time-varying effects of mortality risk factors after the operation. METHODS: We analyzed 11,815 veterans who had coronary artery bypass grafting at any of the 43 Veterans Affairs cardiac surgery centers from October 1997 to September 1999. Time-varying effects of 14 mortality risk factors during the 210 days after coronary artery bypass grafting were evaluated using Cox B-spline regression, which provides an estimate of risk for each variable for each day after operation. RESULTS: Eight variables showed significant time-varying effects after operation. The effect of prior heart operation was very high immediately after operation, but disappeared within 1 week. Three other cardiac variables (prior myocardial infarction, preoperative intraaortic balloon pump, and Canadian Cardiovascular Society anginal class III or IV) also conferred the highest risk on the day of operation and decreased thereafter. In contrast, the four time-varying noncardiac risk variables (age, impaired functional status, chronic obstructive pulmonary disease, and renal dysfunction) showed little or no association with mortality immediately after operation, but had increasing impact during the several months after operation. CONCLUSIONS: A sizable number of mortality risk factors have time-varying effects after coronary artery bypass grafting. Several cardiac risk factors have peak impact immediately after operation but dissipate thereafter. Several noncardiac risk factors confer little risk immediately after operation, but these risks increase during several months. This information may help clinicians focus management strategies for patients during the 7 months after operation. PMID- 12537197 TI - Nonthyroidal illness syndrome in off-pump coronary artery bypass grafting. AB - BACKGROUND: Cardiopulmonary bypass (CPB) is an established cause of nonthyroidal illness syndrome (NTIS). Off-pump coronary artery bypass (OPCAB) has been reported to be less invasive than coronary artery bypass grafting (CABG) with CPB. We prospectively evaluated thyroid metabolism in OPCAB patients. METHODS: We analyzed free thyroid hormones (FT3 and FT4), thyroid-stimulating hormone (TSH), and reverse T3 (rT3) in 20 consecutive patients undergoing CABG surgery. Nine patients underwent CABG with CPB, and 11 underwent OPCAB. Blood samples were taken on admission, on the day of surgery (7:30 AM), after sternotomy, at the end of the operation, and at 2, 6, 12, 24, 36, 48, 72, 96, 120, and 144 hours postoperatively. The concentrations of FT3, FT4, and TSH were determined on each sample. Reverse T3 concentration was measured in 10 patients up to 48 hours and at 144 hours postoperatively. RESULTS: Baseline, operative, and postoperative variables were similar in the two groups. FT3 concentration dropped significantly (p < 0.0001), reaching its lowest value 12 hours postoperatively. There were no significant differences between CPB and OPCAB patients. FT4 varied significantly in both groups (p < 0.0001), but remained in the normal range. TSH variation was not significant. rT3 concentration rose significantly (p = 0.0002) in both groups, peaking 24 hours after surgery. CONCLUSIONS. OPCAB induces a NTIS similar to that observed after CPB, probably due to the inhibition of T4 conversion to T3. This finding suggests that NTIS is a nonspecific response to stress. CPB should not be considered as the sole trigger of NTIS in cardiac surgical patients. PMID- 12537198 TI - Graft of choice to right coronary system in left-sided bilateral internal thoracic artery grafting. AB - BACKGROUND: The complementary graft of choice to the right coronary artery system in patients undergoing left-sided bilateral internal thoracic artery grafting has yet to be determined. Saphenous vein graft (SVG) was compared with right gastroepiploic artery (RGEA) as the supplemental conduit to the right coronary artery when left-sided bilateral internal thoracic artery grafting is implemented. METHODS: From April 1996 to July 1999, 234 patients underwent bilateral internal thoracic artery grafting to the left coronary system with RGEA grafted to the posterior descending artery (RGEA group). They were compared with 127 patients with left-sided bilateral internal thoracic artery in whom SVG was used for grafting the right coronary system (SVG group). RESULTS: Female sex (27% versus 14.5%), diabetic patients (40% versus 27%), emergency cases (21% versus 7.3%), and left main coronary artery disease (34% versus 23%) were more prevalent in the SVG group. Number of grafts per patient was higher in the SVG group (3.8 versus 3.5, p = 0.04). Thirty-day mortality was 3.9% in the SVG and 2.6% in the RGEA group (not significant). Occurrence of postoperative complications (myocardial infarctions, strokes, bleeding, and sternal infections) was similar. Return of angina was similar (1.6% versus 3.8% in the SVG and RGEA groups, respectively). Midterm follow-up (4 to 56 months) showed comparable 1-year and 4 year survival (Kaplan-Meier) for both groups (92.8% and 91.7% in the SVG group, and 94.7% and 88% in the RGEA group, respectively). CONCLUSIONS: In patients undergoing left-sided bilateral internal thoracic artery grafting, the use of RGEA for revascularization of the right coronary system does not confer clinical benefits over SVG after midterm follow-up. PMID- 12537199 TI - Prospective randomized trial of high thoracic epidural analgesia for coronary artery bypass surgery. AB - BACKGROUND: Postoperative pain may be severe after coronary artery bypass surgery. High thoracic epidural analgesia (HTEA) provides intense analgesia. METHODS: Eighty patients were randomized to HTEA or intravenous morphine analgesia (control). Patients received coronary artery bypass surgery (CABG) with cardiopulmonary bypass. Pain was measured by visual analogue scale 0 to 10. Psychologic morbidity, intraoperative hemodynamics, ventricular function, lung function, and physiotherapy cooperation were also assessed. On the third postoperative day HTEA and morphine were ceased and only oral medications were used. Acetaminophen, indomethacin, and tramadol were allowed as supplemental analgesics in both groups. RESULTS: The primary endpoint of pain scores was significantly less with HTEA on postoperative days 1 and 2 at rest, 0.02 +/- 0.2 versus 0.8 +/- 1.8 (p = 0.008) and 0.1 +/- 0.4 versus 1.2 +/- 2.7 (p = 0.022), respectively, and with coughing 1.2 +/- 1.7 versus 4.4 +/- 3.1 (p < 0.001) and 1.5 +/- 2.0 versus 3.6 +/- 3.1 (p = 0.001), respectively. When HTEA and morphine were ceased on day 3, there were no significant differences. The secondary endpoints of postoperative depression (p = 0.033) and posttraumatic stress subscales (p = 0.021) of the Minnesota Multiphasic Personality Inventory were lower with HTEA. Extubation occurred earlier with HTEA, 2.6 versus 5.4 hours (p < 0.001). HTEA showed improved physiotherapy cooperation (p < 0.001), arterial oxygen tension (p = 0.041), and peak expiratory flow rate (p = 0.001). Mean arterial pressure was lower with HTEA (p = 0.036), otherwise there were no differences in intraoperative hemodynamics or ventricular function. CONCLUSIONS: Epidural analgesia reduces pain after coronary operation and is associated with improved physiotherapy cooperation, earlier extubation, and reduced risk of depression and posttraumatic stress. PMID- 12537200 TI - Occlusive wrap dressing reduces infection rate in saphenous vein harvest site. AB - BACKGROUND: Infection in the saphenous vein harvest site is a common problem. We developed an occlusive circumferential wrap dressing technique that reduces skin edge tension, eliminates dead space, and prevents external contamination. We compared the surgical site infection rate using the wrap dressing technique with that of standard longitudinal dressings. METHODS. One hundred fifty-two consecutive patients were randomly assigned to receive either standard dressings or the wrap dressing. Data were collected in the hospital and then 4 to 6 weeks postoperatively. Superficial and deep wound infections were defined by the standard criteria from the Centers for Disease Control and Prevention. RESULTS: The infection rate in the wrap group was 14% compared with 35%, for the standard group (p = 0.006). Multivariate analysis showed that wrap technique was the only significant predictor (negative) of infection (odds ratio, 0.19; p = 0.001). CONCLUSIONS: In saphenous vein harvest wounds, the occlusive wrap dressing technique has the potential to reduce the rate of infection by 50%. This simple and inexpensive technique is also readily applicable to the radial artery harvest site in the arm and may provide similar benefit. PMID- 12537201 TI - Blunt traumatic aortic transection: the endovascular experience. AB - BACKGROUND: Thoracic aortic transection resulting from blunt trauma is usually fatal. It is almost always associated with multiple, complex, nonaortic injuries that could be adversely affected by standard surgical repair of the aorta. Endovascular stenting techniques offer these patients a less physiologically disruptive treatment option. We studied the feasibility and safety of endovascular stent graft placement for treatment of acute traumatic aortic transection. METHODS: Between 1994 and 2001, 9 patients were treated emergently for aortic transections with stent graft placement. The first patient had a custom-made prototype, and the other 8 patients had the Cook-Zenith thoracic stent graft implanted. All were polyester-covered Z-stent construction and deployed through a femoral 20- to 24-F delivery sheath. RESULTS: Stent graft placement successfully sealed the aorta in all patients. One patient died as a result of a cerebrovascular accident. One patient required a brachial thrombectomy to relieve arm ischemia. The remaining eight patients were alive and without complications during the follow-up period (mean 21 months). CONCLUSIONS: Endovascular repair for acute aortic transection is a safe, effective, and timely treatment option. It may be the treatment of choice in patients with extensive associated injuries. PMID- 12537202 TI - Delayed paraplegia after thoracic and thoracoabdominal aneurysm repair: a continuing risk. AB - BACKGROUND: Paraplegia or paraparesis after otherwise successful thoracic or thoracoabdominal aortic reconstruction is a devastating complication for patient and physician. Interventions for its prevention have focused primarily on the intraoperative period. We have recently noted a significant incidence of delayed onset neurologic deficit. METHODS: We reviewed our most recent 5-year experience with thoracic and thoracoabdominal reconstruction to examine the incidence of and potential contributors to delayed paraplegia or paraparesis. RESULTS: Between June 1996 and June 2001, 60 patients (29 men, 31 women) underwent repair of isolated thoracic (n = 26) or thoracoabdominal aortic aneurysm (Crawford I, n = 7; Crawford II, n = 14; Crawford III, n = 12; Crawford IV, n = 1) by the cardiac and vascular surgical services collaboratively. Repair was performed endovascularly in 6, and open with either circulatory arrest in 12, partial left heart bypass in 37, or partial femorofemoral bypass in 5. Operative mortality was 9.3% (5 of 54 patients) for open repair and 0% for endovascular repair. Paraplegia or paraparesis occurred in 6 (10%) patients of which 83.3% (5 of 6) were delayed in onset. All patients with delayed paraplegia or paraparesis had degenerative aneurysms of Crawford extent II (n = 3) or III (n = 2), had intraoperative left heart bypass, and had perioperative spinal drainage. Delayed paraplegia or paraparesis occurred up to 27 days postoperatively, and was associated with a documented episode of hypotension in 60% (3 of 5) of patients. CONCLUSIONS: Improvements in intraoperative management may have reduced immediate paraplegia or paraparesis among vulnerable patients only to leave them at risk of delayed-onset deficit. Postoperative care, including assiduous attention to avoidance of even transient hypotension, must be tailored to this patient population. PMID- 12537203 TI - Total aortic arch replacement through the L-incision approach. AB - BACKGROUND: Even though the median sternotomy is the standard approach for surgery involving the aortic arch, access to the site of distal anastomosis is problematic when the aortic pathology involves the distal arch. We recently developed an "L-incision" approach (a combination of a left anterior thoracotomy and upper half median sternotomy) for total arch replacement. METHODS: We reviewed our surgical technique and operative results for 11 patients who underwent total aortic arch replacement through the L-incision between July 1999 and July 2000. With a patient in a left anterolateral position, a left anterior thoracotomy was performed through the fourth to sixth intercostal space. An upper half median sternotomy followed. Operative exposure was enhanced with spring retractors. The proximal anastomosis (between the four branched graft and ascending aorta) was accomplished first. Upon completion of the proximal anastomosis, the heart was reperfused from one branch of the graft. The three arch vessels were subsequently reconstructed under deep hypothermia and retrograde cerebral perfusion. Antegrade cerebral perfusion was accomplished through the graft as the distal anastomosis (between the graft and descending thoracic aorta) was performed. RESULTS: No early operative deaths were observed. One patient sustained a permanent neurologic deficit. A transient recurrent laryngeal nerve palsy lasting 1 month occurred in 1 patient. No patient required reoperations for bleeding, nor did any patient develop a postoperative phrenic nerve palsy, aspiration pneumonia, or renal dysfunction. CONCLUSIONS: The L incision allows extensive replacement of the aortic arch and is associated with a low incidence of postoperative bleeding and respiratory insufficiency. PMID- 12537204 TI - A long-term experience with the Cabrol root replacement technique for the management of ascending aortic aneurysms and dissections. AB - BACKGROUND: Little information exists regarding the long-term results of the Cabrol technique. This study aimed at exploring, over a 16-year period, the results of aortic root replacements employing this procedure. METHODS: Between 1986 and 2002 forty-five patients (mean age 58.7 +/- 13.8 years old, 84.4% male) underwent a Cabrol procedure. Aortic dissection was the most frequent cause of aortic disease in this series (n = 17), followed by annuloaortic ectasia (n = 10), atherosclerotic aneurysm (n = 5) and poststenotic dilatation (n = 5). Six patients (13.4%) had undergone a previous aortic operation, 8 (17.7%) had a Marfan syndrome and five (11.1%) underwent concomitant arch replacement. Mean clinical follow-up was 87.3 +/- 24.3 months. Twenty-eight patients (93.3% of survivors) had a transesophageal echocardiography (TEE) performed at a mean of 64 +/- 32 months postoperatively. RESULTS: Early mortality was 20%. It was 9.1% for patients with an ascending aortic aneurysm and 41.2% for dissections (p = 0.026). Independent multivariate predictors of early mortality were: aortic dissection (p = 0.009), emergency operation (p < 0.001), operative year (p = 0.02), cross-clamp time (p = 0.001), and CPB duration (p < 0.001). Actuarial survival was 0.77 +/- 0.06, 0.72 +/- 0.06, 0.59 +/- 0.04 and 0.59 +/- 0.04 at 1, 5, 10, and 16 years, respectively. Multivariate analysis revealed age (p = 0.007), cross-clamp time (p = 0.0006) and CPB duration (p = 0.009) as strong predictors of poor late survival. A periprosthetic jet with significant valve regurgitation was detected by TEE in 3 patients. In one of them, an infected periprosthetic space-right ventricular fistula was demonstrated requiring reoperation. Altogether, freedom from reoperation and endocarditis at 16 years was 0.97 +/- 0.02 and 0.94 +/- 0.03, respectively. CONCLUSIONS: The Cabrol technique demonstrated a nonnegligible incidence of early and long-term complications. It should be rarely used and only when a "button" technique is not feasible. PMID- 12537205 TI - Biventricular repair after Norwood palliation. AB - BACKGROUND: In general, neonates with severe left ventricular outflow tract obstruction, aortic valvar stenosis or atresia, and arch hypoplasia with either interruption or coarctation, and a small left ventricle undergo Norwood palliation followed classically by a bidirectional cavopulmonary shunt and eventual modified Fontan. However, a subset of patients, usually neonates with a ventricular septal defect, may have adequate left ventricle and mitral valve sizes making them candidates for future biventricular repair (BVR). In view of the long-term advantage of BVR, the feasibility and outcome of this approach was studied. Additionally, echocardiographic data were reviewed in an attempt to develop objective prognostic criteria for selection of patients suitable for BVR. METHODS: During a 4-year period, 8 of 58 infants undergoing Norwood palliation were identified as potential two-ventricle candidates. Their mean age was 6 days. Diagnoses included aortic atresia (n = 1), or aortic valve stenosis and subaortic stenosis (n = 7), with an interrupted aortic arch in 3 and coarctation in 4. All patients had a ventricular septal defect and a left ventricle that was considered to be apex forming. Mean mitral valve size was 11 mm (z-score = -1.7). Mean aortic valve size was 4.1 mm (mean z-score = -8.4). RESULTS: All 8 patients survived Norwood palliation. Six subsequently underwent BVR with ventricular septal defect closure and a right ventricle to pulmonary artery conduit at a mean age of 7 months. One patient is awaiting repair, and 1 underwent a cavopulmonary shunt. At the time of BVR, mean mitral valve z-score was essentially unchanged at -1.4 (14 mm). No early deaths or late deaths occurred during a mean follow-up of 32 months. CONCLUSIONS: A small subset of patients requiring Norwood palliation as newborns may be candidates for eventual BVR with low risk. In general, patients suitable for BVR have a mitral valve z-score of more than -3 and a normal-sized left ventricle. Recognition of neonatal BVR candidates enables consideration of complete neonatal repair. However, single-stage repair needs to be compared with the excellent results obtainable with the staged approach. PMID- 12537206 TI - Modified Rastelli procedure for double outlet right ventricle with left malposition of the great arteries: report of 9 cases. AB - BACKGROUND: Double outlet right ventricle with L-malposition of the great arteries is a rare type of double outlet right ventricle. This article reviews our experience in treating this disease surgically. METHODS: Between September 1995 and May 1999, 9 patients with double outlet right ventricle [S, D, L] underwent modified Rastelli procedure at Fu Wai Hospital. In all patients, the two great arteries originated completely from the right ventricle. The malformation was associated with subaortic conus and L-transposition of the great arteries. The ventricular septal defect (VSD) was subpulmonary in 3 patients, and non-committed in 6. All but one patient had either pulmonary valvular stenosis or subpulmonary stenosis. No patient had any type of palliative operation before. A right ventriculotomy was made to repair the VSD with a Dacron or vascular prosthesitic patch, and an intraventricular tunnel was made between the left ventricle and the aorta. The main pulmonary artery was divided and the proximal end was closed. A homograft conduit was implanted between the inlet of the right ventricle and the main pulmonary artery. In the Rastelli procedure, the conduit is usually positioned between the right ventricular outflow tract and the pulmonary artery. RESULTS: All patients survived and recovered uneventfully. Echocardiography demonstrated that all intraventricular tunnels and valved conduits were functioning well. The results were satisfactory. CONCLUSIONS: Modified Rastelli procedure is an optimal method for surgically treating double outlet right ventricle with left-malposition of the great arteries. It can completely correct the right ventricular outflow tract stenosis, and right to left shunt, and avoid injuring the right coronary artery. PMID- 12537207 TI - Long-term results after reconstruction of the left ventricular outflow tract by aortoventriculoplasty. AB - BACKGROUND: Aortoventriculoplasty is an established method of reconstruction of complex left ventricular outflow tract (LVOT) obstruction by insertion of a mechanical valve prosthesis after patch enlargement of the LVOT. Little data exist with respect to long-term outcome. METHODS: Between March 1991 and June 2001, 24 patients with a median age of 10.7 (range, 2.1 to 66) years underwent aortoventriculoplasty, which was performed as a primary procedure in 4 and as a secondary intervention in 20 patients. On follow-up, all patients were restudied with an actual prospective evaluation. Data were statistically analyzed using a paired t test. RESULTS: There was one early death from low cardiac output. Four patients had to be reoperated on for bleeding. All 23 postoperative survivors were followed up for 63 (range, 12 to 123) months. Aortic anulus size of 14 (7 to 19) mm could be significantly enlarged to a size of 24 (19 to 27) mm for insertion of a mechanical valve prosthesis (p < 0.001). Blood flow velocity across the LVOT significantly decreased to 1.8 (1.3 to 2.9) m/s (preoperative, 4.1 [2.7 to 5.8] m/s) (p < 0.001). There were no late deaths. One patient underwent late repair of a paraprosthetic leak. On follow-up, there was no hemorrhage related to anticoagulation observed, but there was one minor thromboembolic complication. Relief of LVOT obstruction and good function of the valve prostheses could be demonstrated in all patients. CONCLUSIONS: Aortoventriculoplasty is an easily applicable, low-risk procedure for the effective relief of complex LVOT obstruction, and provides excellent long-term results. PMID- 12537208 TI - The modified Konno procedure for complex left ventricular outflow tract obstruction. AB - BACKGROUND: Complex left ventricular outflow tract (LVOT) obstruction with normal aortic valve function requires aggressive resection in the subaortic region and preservation of the aortic valve. The modified Konno procedure allows generous exposure of the LVOT from the left ventricular apex to the inter leaflet trigones of the aortic valve. Widespread use of this procedure has been limited by concern over injury to the aortic valve, the conduction system, and possibility of residual ventricular septal defect (VSD). METHODS: Retrospective analysis of pertinent data for all patients undergoing the modified Konno procedure (1994 to 2001) at the University of Iowa were reviewed. RESULTS: The modified Konno procedure was used in 18 patients (age 1 to 31) for LVOT obstruction associated with diffuse narrowing of the LVOT (n = 7), a discrete fibrous ring (n = 7), or a fibrous ring associated with abnormal mitral attachments (n = 4). Eight patients had previously undergone LVOT resection. There were no perioperative deaths. Estimated LVOT peak gradients by echocardiogram were 70.4 +/- 24.2 mm Hg (preoperative) and 19.2 +/- 20.4 mm Hg (postoperative) at most recent followup (p < 0.001 vs preop). Aortic insufficiency was moderate in one patient (present preop) and mild or less in all other patients. There were no cases of permanent heart block. Small residual VSDs were present in five patients (28%). Median follow-up is 3.1 years. CONCLUSIONS: The modified Konno procedure can effectively relieve complex LVOT obstruction and preserve aortic valve function. Extension of this procedure for use in the initial presentation of LVOT may be appropriate in cases at increased risk of recurrent LVOT obstruction. PMID- 12537209 TI - Pancreatitis in Fontan patients is related to impaired ventricular relaxation. AB - BACKGROUND: Pancreatitis following cardiopulmonary bypass is a well-known complication in adults undergoing cardiac surgery. However, the occurrence of pancreatitis in pediatric patients undergoing repair of congenital heart disease is under-appreciated. Post-Fontan patients are particularly prone to postoperative pancreatitis. In an effort to identify specific perioperative factors predictive of postoperative pancreatitis, we retrospectively reviewed a group of Fontan patients. METHODS: From June 1996 to June 2001, 40 patients underwent a modified Fontan operation. Four patients developed acute pancreatitis postoperatively. The preoperative, intraoperative, and postoperative hemodynamics and ventricular function parameters were retrospectively analyzed and compared to 10 randomly selected Fontan patients who did not have pancreatitis. Preoperative echocardiographic and angiographic data, including digitized ventricular pressure tracings, were reviewed to obtain ventricular relaxation time constant (tau), pulmonary vascular resistance (PVR), ventricular end diastolic pressure (VEDP), positive dp/dt and negative dp/dt values. RESULTS: Patients developing acute pancreatitis had very high mortality (50%) compared to no mortality in the control group. The mean preoperative tau was significantly prolonged (41 ms vs 26 ms in control group, p < 0.001), and pre operative systemic output (Qs) lower in the pancreatitis group (mean 2.75 L/min/m2) compared with controls (Qs of 5.09 L/min/m2, p < 0.03). CONCLUSIONS: Impaired ventricular relaxation and decreased preoperative cardiac output are predictive of increased risk of postoperative pancreatitis in Fontan patients. Evaluation of preoperative diastolic function in these patients may provide additional insights in to clinical outcome following the Fontan procedure. PMID- 12537210 TI - Stent placement in superior vena cava syndrome. AB - BACKGROUND: Superior vena cava syndrome (SVCS) is often seen in the natural history of malignant thoracic diseases. SVCS is characterized by unpleasant symptoms that usually lead to death. The purpose of our study is to show the efficiency of percutaneous stenting in the superior vena cava for relieving SVCS and the possibility of repeated stenting after recurrence. METHODS: Twenty patients with SVCS caused by malignant diseases who had one or more stents placed in the superior vena cava or its main tributaries were evaluated. RESULTS: Out of 20 patients, 1 died of myocardial infarction 24 hours after the procedure without any signs of pulmonary embolus, hemorrhage, or malposition of the stent. SVCS was successfully controlled in 94% of patients until death or completion of the study. In 3 patients the procedure was repeated (3 to 20 weeks later) because of the recurrence of symptoms. CONCLUSIONS: Percutaneous venous stent placement in the superior vena cava is a simple and effective technique to relieve rapid SVCS caused by malignancies. When recurrence occurs, repeated stenting can be performed successfully. PMID- 12537211 TI - Increased S100B in blood after cardiac surgery is a powerful predictor of late mortality. AB - BACKGROUND: Long-term outcome in patients who suffered stroke after undergoing a cardiac operation has been investigated sparingly, but increased long-term mortality has been reported. S100B is a biochemical marker of brain cell ischemia and blood-brain barrier dysfunction. The aim of this investigation was to record the long-term mortality in consecutive patients undergoing cardiac operations and to explore whether increased concentrations of S100B in blood had a predictive value for mortality. METHODS: Prospectively collected clinical variables, including S100B, in 767 patients who survived more than 30 days after a cardiac operation, were analyzed with actuarial survival analysis and 678 patients were analyzed with Cox multiple regression analysis. RESULTS: Forty-nine patients (6.4%) were dead at follow-up (range, 18 to 42 months); 11.5% (88 of 767 patients) had elevated S100B 2 days after operation (range, 38 to 42 hours). The probability for death at follow-up was 0.239 if the S100B level was more than 0.3 microg/L, and 0.041 if it was less than 0.3 microg/L. The clinical variables independently associated with mortality were preoperative renal failure, preoperative low left ventricular ejection fraction, emergency operation, severe postoperative central nervous system complication, and elevated S100B values, which turned out to be the most powerful predictor. CONCLUSIONS: Even slightly elevated S100B values in blood 2 days after cardiac operation imply a bad prognosis for outcome, and especially so in combination with any central nervous system complication. PMID- 12537212 TI - Beneficial effect of autologous cell transplantation on infarcted heart function: comparison between bone marrow stromal cells and heart cells. AB - BACKGROUND: Cell transplantation may restore function after myocardial infarction, but the optimal cell type remains controversial. We compared autologous bone marrow stromal cells (BMCs) with autologous heart cells (HCs) in a porcine myocardial infarction model. METHODS: Yorkshire pigs underwent coil occlusion of the left anterior descending artery. Bone marrow stromal cells were obtained from sternal marrow and HCs were obtained by left ventricular biopsy, then cultured for 4 weeks. Four weeks after infarction, a 99mTc-sestamibi single photon emission tomography (99mTc-MIBI SPECT) scan was performed and the pigs were then transplanted with BMCs (n = 7), HCs (n = 7), or culture medium (n = 14). Four weeks after transplantation, 99mTc-MIBI SPECT scanning was repeated to evaluate regional perfusion. Pressure-volume loops were constructed from micromanometer and conductance catheter data to evaluate left ventricular function. Hearts were evaluated histologically. RESULTS: Bone marrow stromal cells and HCs engrafted within the infarct and assumed a myocyte morphology. SPECT MIBI scans showed increased perfusion in the infarct in cell-transplanted pigs, while perfusion decreased in the control pigs. Heart cell transplantation improved preload-recruitable stroke work and HC and BMC transplantation both shifted the end-systolic pressure-volume relation to the left. Both BMCs and HCs prevented thinning and expansion of the infarct region, and some BMCs differentiated into endothelial cells in newly formed blood vessels perfusing the infarct. CONCLUSIONS: Both BMCs and HCs engrafted in the infarct region and improved let ventricular function by preventing infarct thinning. Bone marrow stromal cells demonstrated greater plasticity in vivo, and may offer a practical alternative to HC transplantation to restore function and perfusion after a myocardial infarction. PMID- 12537213 TI - Smooth muscle cell hypertrophy of renal cortex arteries with chronic continuous flow left ventricular assist. AB - BACKGROUND: Pathophysiology of long-term continuous flow left ventricular assist is not well described. With many of these devices becoming available, it is important to examine for possible pathologic effects. In this study we examined the relationship between diminished pulsatility and pathologic changes in renal cortical arteries. METHODS: Twenty-nine calves were implanted with various continuous flow left ventricular assist systems in a left ventricle-descending thoracic aorta bypass configuration. Pulsatility was quantified by pulse pressure and pulsatility index. Pathologic changes of the renal cortex arteries were described and evaluated by medial thickness, medial/vascular cross-sectional area ratio, and smooth muscle cell count, to quantify hypertrophy or hyperplasia. Seven calves, which underwent a sham-implant, were used as controls. RESULTS: Systolic arterial pressure, pulse pressure, and pulsatility index were significantly lower and diastolic pressure was significantly higher than before implant in pump-implanted animals. Twenty-three of 29 pumpimplanted calves (79.3%) had medial smooth muscle cell hypertrophy in renal cortex arteries, whereas none of sham-implanted calves had any abnormal lesions. When the pump implanted calves were grouped according to the presence of smooth muscle cell hypertrophy, there was a clear trend toward lower pump flow rate in calves with lesions. Renal function was within the normal range in all calves. CONCLUSIONS: There appears to be a relationship between smooth muscle cell hypertrophy in renal cortex arteries and continuous flow left ventricular assist. Furthermore, although the pathologic changes are likely multifactorial, these lesions appear to be related to lower pump assist rates. PMID- 12537214 TI - Deferoxamine enhances neovascularization and recovery of ischemic skeletal muscle in an experimental sheep model. AB - BACKGROUND: Iron chelators have been reported to interfere with inflammatory cells and possibly enhance vascular growth factor expression. The objective of this study was to investigate the efficacy of the iron chelator deferoxamine mesylate in preventing skeletal muscle ischemia. METHODS: The latissimus dorsi muscle (LDM) was mobilized in 20 adult sheep. Two separate pockets were created in each sheep. Autologous fibrin sealant with or without 100 mg/mL of deferoxamine mesylate (10 pockets) was added to the pockets. Deferoxamine mesylate alone was also applied to another 10 pockets, whereas the 10 other pockets served as controls. RESULTS: Conventional, indirect immunofluorescent enface staining showed that in nonmobilized, nonischemic LDM the capillary density was 196 +/- 14 capillaries/mm2 in the distal and 207 +/- 19 capillaries/mm2 in the middle part. After severe ischemic shock (subtotal mobilization), the muscle did not recover completely even after 2 months (149 +/- 15 capillaries/mm2 in the distal part and 177 +/- 16 capillaries/mm2 in the middle part of the LDM). Fibrin application only increased muscle neovascularization. The number of capillaries per mm2 of muscle increased to 250 +/- 25 in the distal part and to 271 +/- 24 in the middle part of the LDM. However, when fibrin was applied with added deferoxamine mesylate, the capillary density increased to 361 +/- 25 capillaries/mm2 in the distal part (p < 0.05 vs fibrin only; controls) and to 401 +/- 20 capillaries/mm2 in the middle part of the LDM (p < 0.05 vs fibrin only and p < 0.001 vs controls). The data are concordant with the blood flow estimation before and after mobilization (severe ischemic shock) in the different parts of the LDM. CONCLUSIONS: Local application of deferoxamine mesylate enhances neovascularization and recovery of surgically induced skeletal muscle ischemia in a sheep model. PMID- 12537215 TI - Direct compression of the failing heart reestablishes maximal mechanical efficiency. AB - BACKGROUND: In failing hearts, homeostatic mechanisms contrive to maximize stroke work and maintain normal arterial blood pressure at the expense of energetic efficiency. In contrast dobutamine reestablishes maximal mechanical efficiency by promoting energetically optimal loading conditions. However, dobutamine also wastefully increases nonmechanical oxygen consumption. We investigated whether direct mechanical cardiac compression would reestablish maximal mechanical efficiency without the oxygen-wasting effect. METHODS: The pressure-volume relationship and myocardial oxygen consumption were derived in sheep using left ventricular pressure and volume from manometer-tipped and conductance catheters, and coronary flow from Transonics flow probe. RESULTS: Propranolol hydrochloride and atropine sulfate were administered to reduce ejection fraction to 21% when ventricular elastance fell to 1.35 mm Hg/mL and mechanical efficiency to 79% of maximal. Low-pressure direct mechanical compression of the failing heart restored mechanical efficiency to 94% of maximal and realigned optimal left ventricular end-systolic pressure with operating left ventricular end-systolic pressure without altering nonmechanical oxygen consumption. CONCLUSIONS: We conclude that direct cardiac compression restores mechanical efficiency to normal maximum without wasting energy on additional nonmechanical activity. PMID- 12537216 TI - Effect of L-arginine or nitroglycerine during deep hypothermic circulatory arrest in neonatal lambs. AB - BACKGROUND: The role of nitric oxide (NO) in ischemia-reperfusion injury remains controversial. This study evaluated the effects of L-arginine (NO precursor) or nitroglycerine (NO donor) on cardiac and lung function after deep hypothermic circulatory arrest in neonatal lambs. METHOD: Three groups of anesthetized lambs underwent cardiopulmonary bypass, deep hypothermic circulatory arrest (120 minutes at 18 degrees C), and rewarming (40 minutes). During reperfusion, L arginine (5 mg/kg per minute), nitroglycerine (2 microg/kg per minute), or saline (control group) was infused for 100 minutes. All animals were separated from cardiopulmonary bypass and observed for 3 additional hours. Preload recruitable stroke work, cardiac index, pulmonary vascular resistance, alveolar-arterial oxygen difference, and lung compliance plasma nitrate/nitrite levels (NO metabolites) were measured before and after cardiopulmonary bypass. Malondialdehyde in heart tissue and lung tissue was measured 3 hours after cardiopulmonary bypass. RESULTS: Recovery of preload recruitable stroke work and cardiac index were significantly higher in the L-arginine and nitroglycerine groups than in the control group (p < 0.05). Pulmonary vascular resistance was significantly lower in the L-arginine and nitroglycerine groups than in the control group (p < 0.05). Levels of NO metabolites and issue malondialdehyde did not differ among groups. CONCLUSIONS: L-arginine and nitroglycerine improved recovery of left ventricular function and reduced pulmonary vascular resistance after deep hypothermic circulatory arrest. The mechanism of beneficial action could involve increased NO levels, but we did not find higher levels of NO metabolites compared with controls. Tissue malondialdehyde levels were not affected by L-arginine or nitroglycerine. These results show that, at these dosage levels, provision of substrate for NO production or provision of an NO donor were beneficial to the recovery of myocardial and pulmonary vascular function. PMID- 12537217 TI - Therapeutic angiogenesis using autologous bone marrow stromal cells: improved blood flow in a chronic limb ischemia model. AB - BACKGROUND: We evaluated the effect of autologous marrow stromal cells (MSCs) on neovascularization and blood flow in an animal model of chronic limb ischemia. METHODS: Chronic hind limb ischemia was created by ligating the left common iliac artery of male Lewis rats. Three weeks after ligation, 5.0 million LacZ+MSCs (n = 10) or culture medium (n = 10) were injected into the anteromedial muscle compartment of the left thigh. At 4 and 6 weeks after injection, half the animals (n = 5) from each group underwent femoral artery ultrasonic blood flow measurements of the ischemic and nonischemic limbs to obtain a flow ratio. The animals also underwent angiography and measurements of blood vessel density and arteriolar density. Qualitative histologic assessment of the limb muscles was performed. RESULTS: LacZ+MSCs were found to differentiate into endothelium (F VIII+), vascular smooth muscle (positive a-smooth muscle actin), skeletal muscle (positive desmin), and adipocytes. Ischemic hind limbs where MSCs were implanted had greater vascular density and arteriolar density than control limbs (p < 0.001). Femoral artery flow index (left femoral artery flow/right femoral artery flow) was 0.89 +/- 0.12 and 0.90 +/- 0.06 for rats injected with MSCs measured at 4- and 6-weeks, respectively, compared with 0.50 +/- 0.15 and 0.50 +/- 0.10 for the control rats (p < 0.001). Angiography demonstrated reconstitution of the left femoral artery in rats that received MSC implantation through pelvic and abdominal wall collateral formation. CONCLUSIONS: Local MSC implantation induces a neovascular response resulting in a significant increase in blood flow to the ischemic limb. Marrow stromal cells are also capable of spontaneously regenerating the various components of muscular tissues. PMID- 12537218 TI - Aprotinin inhibits leukocyte-endothelial cell interactions after hemorrhage and reperfusion. AB - BACKGROUND: The serine protease inhibitor aprotinin has been successfully used to reduce blood loss in patients undergoing cardiac operations. We studied aprotinin for its ability to modulate leukocyte-endothelial cell interactions after ischemia and reperfusion. METHODS: The effects of aprotinin on leukocyte endothelial cell interactions were observed by intravital microscopy in the rat mesenteric microcirculation and immunohistochemical analysis. The inflammatory cascade (leukocyte rolling, firm adherence, and transmigration) was studied after thrombin stimulation and after hemorrhage and reperfusion. RESULTS: Intravenous bolus administration of aprotinin treatment (20,000 U/kg) significantly reduced leukocyte rolling from 55 +/- 8 to 17 +/- 3 cells/min (p < 0.01) and adherent cells from 12 +/- 2 to 7 +/- 1.4 cells (p < 0.05) along the venous endothelium of the rat mesentery after thrombin activation. In addition, aprotinin pretreatment significantly inhibited transmigration of leukocytes from 11.3 +/- 1.2 to 6.0 +/- 1.1 cells (p < 0.05) through the microvascular endothelial wall. Similarly, aprotinin decreased leukocyte-endothelium interaction after hemorrhagic shock. Moreover, immunohistochemistry demonstrated that aprotinin significantly attenuated P-selectin expression by the intestinal vascular endothelium. CONCLUSIONS. Our data demonstrate that aprotinin potently inhibits recruitment of leukocytes in the microvasculature by interfering with endothelial cell polymorphonuclear neutrophil interaction, and is a potent endothelial protective agent in clinically relevant doses. Thus, aprotinin pretreatment may be useful for primary prevention of inflammatory tissue injury mediated by ischemia reperfusion injury such as shock, trauma, open heart operation, or other extensive vascular surgical procedures. PMID- 12537219 TI - Outcomes after esophagectomy: a ten-year prospective cohort. AB - BACKGROUND: The Department of Veterans Affairs National Surgical Quality Improvement Program is a unique resource to prospectively analyze surgical outcomes from a cross-section of surgical services nationally. We used this database to assess risk factors for morbidity and mortality after esophagectomy in Veterans Affairs Medical Centers from 1991 to 2001. METHODS: A total of 1,777 patients underwent an esophagectomy at 109 Veterans Affairs hospitals with complete in-hospital and 30-day outcomes recorded. Bivariate and multivariable analyses were completed. RESULTS: Thirty-day mortality was 9.8% (174/1,777) and the incidence of one or more of 20 predefined complications was 49.5% (880/1,777). The most frequent postoperative complications were pneumonia in 21% (380/1,777), respiratory failure in 16% (288/1,777), and ventilator support more than 48 hours in 22% (387/1,777). Preoperative predictors of mortality based on multivariable analysis included neoadjuvant therapy, blood urea nitrogen level of more than 40 mg/dL, alkaline phosphatase level of more than 125 U/L, diabetes mellitus, alcohol abuse, decreased functional status, ascites, and increasing age. Preoperative factors impacting morbidity were increasing age, dyspnea, diabetes mellitus, chronic obstructive pulmonary disease, alkaline phosphatase level of more than 125 U/L, lower serum albumin concentration, increased complexity score, and decreased functional status. Intraoperative risk factors for mortality included the need for transfusion; intraoperative risk factors for morbidity included the need for transfusion and longer operative time. CONCLUSIONS: These data constitute the largest prospective outcomes cohort in the literature and document a near 50% morbidity rate and 10% mortality rate after esophagectomy. Data from this study can be used to better stratify patients before esophagectomy. PMID- 12537220 TI - Low-dose oral amiodarone prophylaxis reduces atrial fibrillation after pulmonary resection. AB - BACKGROUND: Atrial fibrillation after pulmonary resection increases morbidity and costs. To evaluate the efficacy of low-dose oral amiodarone (LDOA) as prophylaxis for atrial fibrillation after pulmonary resection, we reviewed all patients 60 years or older having pulmonary resections by thoracotomy in a 30-month period. METHODS: We identified 31 patients who received prophylactic LDOA (200 mg by mouth every 8 hours) while hospitalized and 52 patients who received no prophylactic treatment. The groups were comparable for sex, age, comorbidities, and surgical procedure. RESULTS: Twenty of 83 patients (24%) had postoperative atrial fibrillation: 17 of 52 patients (33%) without prophylaxis and 3 of 31 (9.7%) with prophylaxis (odds ratio, 0.221; 95% confidence interval, 0.059 to 0.829; p = 0.0253). The median total hospital charge was $30,800 (range, $20,400 $96,900) for 50 patients without prophylaxis and $26,700 (range, $11,000-$55,900) for 31 patients with prophylaxis (p = not significant). Patients receiving LDOA had lower accumulated charges per day of hospital stay (p = 0.0011). CONCLUSIONS: LDOA prophylaxis significantly reduces the incidence of atrial fibrillation after pulmonary resection. Its use in this population may be cost-effective. Results of this pilot study provide a rationale for a prospective randomized trial. PMID- 12537221 TI - Mediastinal lymph node staging in suspected lung cancer: comparison of positron emission tomography with F-18-fluorodeoxyglucose and mediastinoscopy. AB - BACKGROUND: In patients with bronchogenic carcinoma, mediastinal lymph node staging is essential for determining treatment options. In this retrospective analysis we compared the results of positron emission tomography (PET) using F-18 fluorodeoxyglucose with those of mediastinoscopy in nodal staging for suspected bronchogenic carcinoma. METHODS: From March 1997 to June 2001, 102 patients (86 male,16 female, age 62 +/- 9 years) underwent both PET and mediastinoscopy for radiologically suspected mediastinal lymph node disease in bronchogenic carcinoma. Total body emission scans were acquired 90 to 150 minutes after injection of 230 MBq of F-18 fluorodeoxyglucose. Mediastinoscopic evaluation of lymph node stations was performed according to the method of Mountain and Dresler (1R, 1L, 2L, 2R, 4L, 4R,7). Patients were eligible if surgical staging was performed within 6 weeks after the PET scan. RESULTS. Of the 102 patients, benign lesions were diagnosed in 15. In 87 patients malignant disease was proven by histology, and bronchogenic carcinoma was found in 82. Of 469 nodal stations analyzed, malignancy was documented by histology in 84. In PET analysis 79 true positive and 304 true-negative samples were found. Five lymph node stations were false negative, and 81 samples were false positive. False-positive findings in PET frequently were seen in inflammatory lung disease. The sensitivity of PET was 94.1%, specificity was 79% with a diagnostic accuracy of 81.6%. The positive predictive value of PET was 49.3%, and the negative predictive value was 98.4%. CONCLUSIONS: In patients with positive PET scan results histologic verification appears necessary for exact lymph node staging. In view of the negative predictive value mediastinoscopy can be omitted in patients with bronchogenic carcinoma whose PET scan results were negative. PMID- 12537222 TI - Limited resection for non-small cell lung cancer: observed local control with implantation of I-125 brachytherapy seeds. AB - BACKGROUND: Limited resection for lung cancer has been associated with a relatively high incidence of local recurrence. This retrospective study evaluates the impact of implanting radioactive iodine-125 (125I) seeds along the resection margin in these patients. METHODS: Thirty-three patients with lung cancer who were not candidates for lobectomy or pneumonectomy underwent a limited resection of 35 primary non-small cell lung cancers. 125I brachytherapy seeds were implanted along the resection margin to reduce the risk of local recurrence. Survival using the Kaplan-Meier method and sites of recurrence were documented. Follow-up ranged from 20 to 98 months (median, 51 months). RESULTS: The 5-year survival was 47% for all patients. For patients with T1N0 tumors, it was 67%, and for patients with T2N0 tumors, it was 39%. However, the cancer-specific survivals were 77% and 53% for patients with T1N0 and T2N0 tumors, respectfully. Ten patients experienced recurrence, with two local (at the resection margin) and six regional recurrences (five mediastinum, one chest wall). Both local recurrences and one regional recurrence occurred in the 19 patients with T1N0 tumors. CONCLUSIONS: 125I seed implantation along the resected margin for compromised patients undergoing limited resection of lung cancer results in a relatively low incidence of local recurrence and may prolong survival. PMID- 12537223 TI - Metastatic thoracic lymph node carcinoma with unknown primary site. AB - BACKGROUND: Metastatic cancer in thoracic lymph nodes without a primary site is rare. The purpose of this study is to draw attention to this probably underestimated entity, to speculate on its possible origins, and to suggest guidelines for its treatment. METHODS: Eight heavy smokers with no past medical history of cancer were diagnosed at operation to have malignant cells in intrathoracic lymph nodes (N1 or N2) with no primary site in the lung. All patients underwent an exploratory thoracotomy with a presumed diagnosis of lung cancer except one who presented with a middle lobe mucosa-associated lymphoid tissue lymphoma. We reviewed the type of surgical resection, histologic and immunohistochemical analysis of resected specimens, treatments, survival, and long-term results. RESULTS: Resections performed were pneumonectomy (n = 4), lobectomy (n = 3), and bilobectomy (n = 1). All patients underwent complete mediastinal lymph node dissection. Lung resection was performed for mucosa associated lymphoid tissue lymphoma (n = 1) and for tumorlike lesions that appeared to be tuberculoma (n = 1) and intrapulmonary metastatic lymph nodes (n = 6). Malignant cells were located in intrapulmonary lymph nodes alone (n = 3) or also in mediastinal lymph nodes in three other cases. All these tumors were cytokeratin-positive, demonstrating their epithelial nature. Pulmonary origin was confirmed in two cases (thyroid transcription factor 1-positive and thyroglobulin negative). No other origin could be demonstrated by immunochemistry. Three patients died within the first year. All other patients are still alive without recurrence (Kaplan-Meier 5-year survival rate, 62.5%). CONCLUSIONS: Frequency of metastatic cancer in thoracic lymph nodes without a primary site is probably underestimated because the cancer is routinely diagnosed by mediastinoscopy and considered as metastatic disease not amenable to operation. The origin of the disease, either pulmonary, endogenous, or from extrathoracic sites, is often difficult to assess. Nevertheless, our data confirm those of the literature and demonstrate that survival can be increased by operation. This implies diagnosis of the entity and consideration that thoracic lymph node involvement can apparently be isolated and therefore resectable. PMID- 12537224 TI - Outcome at 2 years of infants with congenital diaphragmatic hernia: a population based study. AB - BACKGROUND: Management of neonates with congenital diaphragmatic hernia (CDH) has undergone many changes associated with increased survival of high-risk CDH. However, little is known about the long-term outcome of CDH infants. METHODS: Follow-up was performed in 85 newborn infants with CDH admitted in our neonatal intensive care unit between January 1991 and December 1998. Early (< 2 months) and late mortality (> or = 2 months), and respiratory, nutritional, musculoskeletal, and neurosensory outcome at 2 years were recorded. RESULTS: Surgical repair was performed in 59 infants (69%) at a median postnatal age of 124 (range, 38 to 246) hours. Extracorporeal membrane oxygenation was used in 26 (30%) newborn infants. Survival at 2 years was 51 of 85 (60%) (early death, 28/85 [33%]; late death, 6/85 [7%]). Late deaths occurred because of persistent pulmonary hypertension or iatrogenic complications. Twelve of 51 (24%) newborn infants were oxygen dependant at the postnatal age of 28 days, and 1 of 51 (1.9%) was still oxygen dependant at 2 years. Growth failure was noted in 9 of 51 (18%), mainly related to severe gastroesophageal reflux and oral aversion. Scoliosis was diagnosed in 2 infants. Neurologic examination at 2 years was normal in 45 of 51 (88%). Cerebral palsy and developmental delay were observed in 2 and 4 infants, respectively. Four infants (8%) experienced associated problems. Respiratory, nutritional, and musculoskeletal morbidity was higher in infants treated by extracorporeal membrane oxygenation (p < 0.05). CONCLUSIONS: CDH infants are at risk for adverse nutritional and respiratory outcome. Despite severe respiratory failure at birth, prolonged oxygen therapy above 2 years of age is uncommon. Conversely, failure to thrive related at least in part to gastroesophageal reflux and oral dysfunction remains the major problem at 2 years of age. However, both nutritional and respiratory problems tend to improve with age. PMID- 12537225 TI - Intrathymic inoculation of donor bone marrow induces long-term acceptance of lung allografts. AB - BACKGROUND: We investigated whether intrathymic inoculation of donor bone marrow at the time of transplantation induced long-term acceptance of lung allografts. METHODS: Four- to-six-week-old August Copenhagen Irish (ACI) and Wistar Furth (WF) rats were used as donors and recipients, respectively. After being inoculated intrathymically with either donor-specific (ACI) or third-party (F344) bone marrow (2.0 x 10(7) cells/lobe), the recipient (WF) animal received a left lung transplant from an ACI donor. A short course of tacrolimus (1 mg/kg per day for 5 days) was administered. Animals were sacrificed at timed intervals after transplantation, and rejection was graded on a scale of 0 (none) to 4 (severe). RESULTS: At 28 days, animals receiving donor-specific bone marrow have lower (p < 0.01) median rejection grade (MRG = 0.25; n = 6) than those receiving third-party bone marrow (MRG = 3; n = 6) and controls (no bone marrow; MRG = 2.5; n = 6). Animals receiving intrathymic donor bone marrow accepted lung allografts up to 380 days with minimal rejection (MRG = 2; n = 6). Long-term lung recipients also accepted a challenging donor-specific heart graft (n = 4) for more than 150 days. In mixed lymphocyte reaction assays, T lymphocytes of WF recipients that had received intrathymic bone marrow (from ACI donor) exhibited low response (similar to self antigens) to donor (ACI) cells, but reacted strongly (five times higher) to third-party (F344) cells. CONCLUSIONS: Intrathymic inoculation of donor bone marrow at the time of transplantation along with a short course of tacrolimus induces long-term acceptance of lung allografts in rats. This simple approach of tolerance induction may have clinical application. PMID- 12537226 TI - Favorable outcome with bivalirudin anticoagulation during cardiopulmonary bypass. AB - An 81-year-old man with previous syncopal episodes, progressive shortness of breath, pulmonary edema, severe calcific aortic stenosis, and a history of heparin-induced thrombocytopenia required aortic valve replacement. Bivalirudin, a thrombin-specific anticoagulant, was used in place of heparin. The patient received a 50 mg bivalirudin bolus followed by an infusion between 1.5 mg x kg( 1) x h(-1) and 1.75 mg x kg(-1) x h(-1). Adequate anticoagulation was readily obtained resulting in an uneventful cardiopulmonary bypass. Activated clotting time (ACT) values steadily declined after discontinuation of the bivalirudin infusion. Bivalirudin is a practical alternative to heparin during cardiac surgical procedures. PMID- 12537227 TI - Isolated cerebral perfusion for intraoperative cerebral malperfusion in type A aortic dissection. AB - Cerebral malperfusion due to expansion of a false lumen can occur acutely during aortic repair when retrograde femoral perfusion is initiated. We detected this catastrophe by a rapid decrease in regional cerebral oxygenation and successfully treated it by immediate isolation of the cerebral circulation from the systemic circulation. The surgical management, including the above technique, for this rare event is described. PMID- 12537228 TI - Late quadriplegia after adult coarctation repair. AB - Quadriplegia developed suddenly 1 month after coarctation repair in a 53-year-old human. Cervical cord ischemia caused by thrombosis in an enlarged anterior spinal artery collateral was diagnosed on magnetic resonance scan and spinal angiography. After urgent cord decompression and anticoagulation there was neurologic improvement over a period of several months. Urgent investigation of neurologic abnormalities occurring late after coarctation repair may enable intervention to avert permanent neurologic sequelae. PMID- 12537229 TI - Internal mammary artery steal in a dialysis patient. AB - A 67-year-old man with chronic renal failure who was being treated with dialysis through an arteriovenous fistula in the left arm was scheduled to undergo off pump coronary artery bypass grafting. Angiography detected the steal phenomenon in the left internal mammary artery during the diastolic phase. Postoperative study with Doppler guidewire showed that the steal of the left internal mammary artery was due to an ipsilateral arteriovenous fistula. Preoperative angiography of the internal mammary artery is therefore recommended for patients on dialysis. PMID- 12537230 TI - The fenestrated Kawashima operation for single ventricle with interrupted inferior vena cava. AB - An 8-month-old boy with double outlet right ventricle with hypoplastic left ventricle, heterotaxy, left atrial isomerism, bilateral superior vena cavae without bridging vein, and interruption of the inferior vena cava with azygous continuation to the left superior cava underwent a bilateral bidirectional cavopulmonary anastomosis. A calibrated 3-mm connection between the right pulmonary artery and the common atrium was constructed with the proximal right superior vena cava to allow right to left shunting, analogous to a fenestration in a Fontan operation. We hypothesize that in small young patients undergoing the Kawashima operation a fenestration may improve postoperative hemodynamics. PMID- 12537231 TI - Coil embolization of a patent ductus venosus in a 52-day-old girl with congenital heart disease. AB - We present the case of 52-day-old girl with a common atrioventricular canal. Severe liver dysfunction persisted following complete repair of the cardiac defect. A patent ductus venosus appeared to be the source of the hemodynamic disturbance responsible for hepatic dysfunction. Given her critical condition, coil embolization of the ductus venosus was performed, after which the patient improved rapidly. The ductus venosus should be tested for patency when liver dysfunction persists after the corrective cardiac surgery, and coil embolization is the treatment of choice in gravely ill children. PMID- 12537232 TI - Acquired pulmonary vein stenosis as a cause of life-threatening pulmonary hypertension. AB - We report the case of an infant born prematurely at 27 weeks gestational age with life-threatening pulmonary hypertension crisis as a result of left upper pulmonary vein stenosis. Surgical treatment consisted of a lobectomy, which is a safe and effective procedure. Evidence strongly suggests that the venous stenosis may have resulted from hypertonic drugs infused through an umbilical catheter facing the upper left venous-atrial junction. PMID- 12537233 TI - Bilateral pulmonary artery banding for resuscitation in hypoplastic left heart syndrome. AB - We report a case of a hypoplastic left heart syndrome with a nearly intact atrial septum and an obstructed anomalous pulmonary to systemic venous connection. Surgical atrial septectomy followed by bilateral pulmonary artery banding provided an optimal condition for the Norwood operation. PMID- 12537234 TI - Absent pulmonary valve syndrome with intact ventricular septum and patent ductus arteriosus: report of two cases and a short review of the literature. AB - We describe two patients who both presented shortly after birth with congestive heart failure due to an absent pulmonary valve and patent ductus arteriosus. Diagnostic evaluation revealed in both cases an aneurysmatic dilation of the pulmonary vascular tree and an abundant left-to-right shunt over a large patent ductus arteriosus. Ventricular septal defects were not detected. Early surgical closure of the patent ductus arteriosus improved the hemodynamic situation so that implantation of a homograft valve could be delayed. PMID- 12537235 TI - Heart transplant for anomalous origin of left coronary artery from pulmonary artery. AB - Anomalous origin of the left coronary artery from the pulmonary artery is a congenital coronary artery malformation most commonly present in infancy. A variety of surgical procedures have been described to achieve physiological correction of the coronary flow abnormalities. These techniques are effective as long as there is potential for myocardial recovery. However the sequelae of chronic myocardial ischemia that characterize this entity often irreversibly damage the heart and preclude correction and palliation of the native anomaly. In this type of setting, heart transplantation is a realistic option. Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) occasionally presents in adulthood. Anatomic repair with a two coronary artery system may not be optimal in patients presenting with ischemic cardiomyopathy. We report an adult patient with platelet factor 4 (PF4) antibodies who underwent orthotopic heart transplantation (OHT) for ALCAPA. PMID- 12537236 TI - Late-onset chylothorax after heart-lung transplantation. AB - Chylothorax has rarely been reported after heart and lung transplantation. We describe a case of a bilateral chylothorax that occurred in a 53-year-old man 4 months after heart-lung transplantation and review the three additional reports in the literature of chylothorax after heart transplantation. Chyle drainage may cause compromise of the transplant patient. The good nutritional status of our recipient, however, allowed conservative therapy with chest tube drainage and enteral low-fat diet, without the need for surgical intervention. We concluded that chylothorax can presents late after lung transplantation and that conservative treatment should be considered in patients with good nutritional status. PMID- 12537237 TI - Successful treatment of Novacor pump pocket infection by omental transposition. AB - Device-related infection remains a major factor restricting the long-term use of left ventricular assist systems. Severe pocket infection is especially difficult to manage and removal of the device has been the only curative treatment in most cases. We report a case of a Novacor device pocket infection treated successfully with continuous local irrigation and transposition of omental flap. This procedure provides another option for the management of pocket infection, which is mandatory for destination therapy the permanent usage of LVAS for the purpose of circulatory supports in patients with end-stage heart failure, who are not indicated for heart transplantation. PMID- 12537238 TI - Esophageal-pericardial fistula with purulent pericarditis secondary to esophageal carcinoma presenting with tamponade. AB - A case of esophago-pericardial fistula secondary to esophageal carcinoma causing pericardial effusion and tamponade is presented. Palliation can be achieved effectively by limited thoracotomy, pericardial resection and drainage, and in selected cases esophageal stenting. PMID- 12537239 TI - Modified Dumon stent for the treatment of a bronchopleural fistula after pneumonectomy. AB - We describe the case of a 72-year-old man in whom a bronchopleural fistula developed after induction chemotherapy followed by right pneumonectomy for lung cancer. Resuturing of the right main bronchial stump, endoscopic treatment, and repair using a latissimus dorsi muscle flap were not effective. Consequently we placed a modified Dumon stent in the carina, which effectively closed the stump. PMID- 12537240 TI - Giant hydatid cysts of lung and liver. AB - A 13-year-old girl was admitted to our clinic with the complaints of dyspnea, abdominal distention, cough, and right-sided chest pain.The diagnosis of disease was made with physical examination and radiologic evaluation. She underwent right thoracotomy and phrenotomy for the management of the hydatid cysts of the lung and the liver. The postoperative course was uneventful and she was discharged on the 10th postoperative day. We believe that the simultaneous management of concomitant pulmonary and hepatic hydatid cysts through a thoracicroute is a convenient option. PMID- 12537241 TI - Delayed para-aortic hematoma after Bentall operation. PMID- 12537242 TI - Radical resection of a giant chondrosarcoma of the anterior chest wall. PMID- 12537243 TI - Stapled explant of the Jarvik 2000 left ventricular assist device. AB - Currently the most common indication for placement of a left ventricular assist device is as a bridge to heart transplantation. One of the new generation axial flow left ventricular assist devices is the Jarvik 2000. This device is placed in the apex of the left ventricle and the outflow graft passes through the left pleural space and is anastomosed to the descending thoracic aorta. The course of the outflow graft presents technical challenges during explant for heart transplantation. Opening the posterior pericardium and use of a vascular stapler to control the outflow graft at the level of the descending thoracic aorta facilitates easy explantation. PMID- 12537244 TI - Modified technique for mitral repair without ring annuloplasty. AB - Mitral valve repair is the procedure of choice to correct mitral regurgitation. Most operative techniques use an annuloplasty ring to provide stability and durability to the correction. We present a modification of existing repair techniques, without the use of an annuloplasty ring, in which plication sutures allow both annular remodeling and stability. Clinical and echocardiographic follow-up in our series of 60 patients with a mean follow-up of 29 months is presented. PMID- 12537245 TI - Modified Daggett's technique for early repair of postinfarct posterior ventricular septal rupture. AB - A surgical modification for safe early repair of posterior septal rupture is described. This technique is based on the method described by Daggett, but adds one internal patch, plus the application of fibrin glue between the internal and external patch for minimizing bleeding. This modification is a simple and reliable one for repairing posterior ventricular septal rupture. PMID- 12537246 TI - Pulmonic valve annular enlargement with valve repair in tetralogy of Fallot. AB - We present an operative technique of pulmonic valve annular enlargement with concomitant valve repair using two pericardial patches to reduce pulmonary regurgitation after complete repair of tetralogy of Fallot. We have used this technique in 18 patients with tetralogy of Fallot with excellent results. PMID- 12537247 TI - Off-pump coronary artery bypass grafting: not a flash in the pan. AB - During the past decade, technical improvements have made off-pump coronary artery bypass operations a routine procedure. During this time, off-pump coronary artery bypass has been audited against conventional techniques by many observational, case-matched, and prospective randomized studies. There is evidence in the literature suggesting that off-pump coronary artery bypass operations reduce postoperative morbidity, organ dysfunction, and costs, without compromising midterm outcome compared with conventional coronary operations. The available evidence also supports the view that high-risk patients might benefit the most from revascularization on the beating heart. High quality follow-up data are still needed to assess the impact of off-pump coronary artery bypass operations on long-term clinical outcome. PMID- 12537248 TI - Depression and anxiety and outcomes of coronary artery bypass surgery. AB - Psychological and psychiatric disorders independently increase the risk of cardiovascular disease and worsen the prognosis in patients with established cardiovascular lesions. The objective of this literature review is to discuss recent data concerning the relationships between depression and anxiety and the outcomes of coronary artery bypass grafting. Pathophysiological hypotheses are put forward to explain observed links. We suggest recommendations aimed at improving the psychological evaluation and management of heart surgery candidates, as well as postbypass patients, in the hope of improving quality of life and cardiovascular outcomes in these patients. PMID- 12537249 TI - Can any conclusion be justified with a 27% loss of patients at follow-up? PMID- 12537250 TI - Descending aorta-obtuse marginal coronary artery bypass grafting using an automatic anastomotic device. PMID- 12537251 TI - One-stage repair of complete atrioventricular canal, double-outlet right ventricle, left superior vena cava, cor triatriatum and pulmonary stenosis. PMID- 12537252 TI - Ultraflex expandable metallic stents: difficulties in benign tracheal stenoses. PMID- 12537253 TI - Mammary leptin synthesis, milk leptin and their putative physiological roles. AB - This paper reviews data on mammary leptin and leptin receptor gene expression as well as on blood and milk leptin levels during the pregnancy-lactation cycle in humans, rodents and ruminants, with the aim of better understanding milk leptin origin and functions. The few published papers report that leptin may be produced by different cell types in the mammary tissue, and may act as a paracrine factor on mammary epithelial cell proliferation, differentiation and/or apoptosis via adipose-epithelial and/or myoepithelial-epithelial cellular interactions. In addition to leptin synthesis, epithelial cells may transfer leptin from the blood, and these two mechanisms may account for the presence of leptin in the milk. The respective parts of these two processes remain to be determined, as well as the true milk leptin levels. Indeed, reported concentrations for milk leptin vary strongly according to species and mainly according to the milk fractions and the assay methods used. If leptin levels in milk (and specially colostrum) are found to be significant, this hormone could be involved in neonate physiology. PMID- 12537254 TI - Muscle fibre ontogenesis in farm animal species. AB - In farm animals (bovine, ovine, swine, rabbit and poultry), muscle fibre characteristics play a key role in meat quality. The present review summarises the knowledge on muscle fibre characteristics and ontogenesis in these species. Myofibre ontogenesis begins very early during embryonic life, with the appearance of two or three successive waves of myoblasts which constitute the origin of the different types of muscle fibres. In small animals (rodents and poultry), a primary and a secondary generation of fibres arise respectively during the embryonic and foetal stages of development. In the largest species (bovines, sheep, pigs) a third generation arises in the late foetal or early postnatal period. Following these two or three waves of myogenesis, the total number of fibres is fixed. This occurs during foetal life (bovines, ovines, pigs and poultry) or during the first postnatal month in rabbits. Contractile and metabolic differentiation proceed by steps in parallel to myogenesis and are partially linked to each other. In bovines and ovines, the main events occur during foetal life, whereas they occur soon after birth in the pig, poultry and rabbit, but some plasticity remains later in life in all species. This comparative survey shows that the cellular processes of differentiation are comparable between species, while their timing is usually species specific. PMID- 12537255 TI - The impact of genetic polymorphisms on the protein composition of ruminant milks. AB - The purpose of this review is to give an overview of our current knowledge on the polymorphisms occurring in genes coding for milk proteins and responsible for quantitative variability in their expression, thus influencing the protein composition of livestock ruminant milk. The overall genomic organisation of the 6 main ruminant milk protein genes: alpha-lactalbumin, beta-lactoglobulin and the four caseins (alpha(s1), alpha(s2), beta and kappa), their chromosomal location and their expression pattern are first summarised before presenting general mechanisms controlling gene expression both at the transcriptional and the post transcriptional levels. Polymorphisms found in cis-regulatory elements, mainly within the 5'-flanking region of the genes encoding beta-lactoglobulin and alpha(s1)- and alpha(s2)-caseins, have been found, in cattle, to influence their transcription rate. In addition, polymorphisms found in the transcription unit, within intron as well as exon sequences, have been shown to be responsible for defects in the processing of primary transcripts and/or the export of messenger RNA to the cytoplasm. Mutations responsible for the occurrence of premature stop codons in alpha(s1)- and beta-casein mRNAs have been shown to be associated both with a decrease in the level of the relevant transcripts and the existence of multiple forms of messengers due to alternative splicing (exon skipping, usage of cryptic splice sites). Such a situation, well-exemplified by the gene encoding alpha(s1)-casein in the goat, may have dramatic biological consequences (secretion pathway, casein micelle structure, fat content, etc.) by modifying the message and accordingly the primary structure of the protein as well as its expression. Since some of these polymorphisms dramatically affect technological properties of milk, including cheese yields and organoleptic characteristics, methods mainly based on the PCR technique have been designed and applied in selection and breeding programmes to improve milk protein quality. PMID- 12537257 TI - Osteoarthritis: biological markers for the future? PMID- 12537256 TI - Bioactive substances of plant origin in food--impact on genomics. AB - In the past decade, substantial progress has been made concerning our knowledge of bioactive components in plant foods and their links to health. Human diets of plant origin contain many hundreds of compounds which cannot be considered as nutrients, but appear to play a role in the maintenance of health. These substances are called nutraceuticals. In some cases where the disease process is at least partially understood, elements of protection can be related to a single compound or structurally related group of compounds in the diet. Bioactive components of food which are of special interest include the following groups: polyphenols, phytoestrogens, phytosterols, phytates and polyunsaturated fatty acids. Most of them are featured by antioxidant properties. In the first part of this review, we indicate the main groups of bioactive compounds giving a description of their localisation, chemical properties and biological actions. Recently, it was shown, however, that the bioavailability of potential antioxidants from plant foods is generally too low to have any substantial direct effect on reactive oxygen species. As a result of that it is postulated that dietary compounds, even in very low concentrations, may have a far greater impact than previously appreciated on the regulation of gene expression. The second part of this paper concerns the action of the literally most important bioactive substances on the molecular mechanisms of the control of genes which in turn affect cellular metabolism. A few current studies on the action of selected nutraceuticals on the activity of transcription factors such as AP-1, NF-kappaB, SREBPs, PPARs as final targets in the signal transduction cascade and gene regulation are included. A detailed analysis of numerous factors of dietary origin with their targets is far beyond the scope of this paper. However, continuing research on the effects of nutraceuticals on gene expression should provide insight into the mechanisms of prevention of diseases such as obesity, diabetes, atherosclerosis, hypertension and cancer by dietary manipulations. PMID- 12537258 TI - Are antidepressants effective in fibromyalgia? AB - Amitriptyline and other imipramine antidepressants are the cornerstone of drug therapy in fibromyalgia. However, some patients fail to respond to antidepressant therapy, and in responders the beneficial effects wear off after some time. The effect on fibromyalgia seems independent from the effects on depression. PMID- 12537259 TI - Prescribing outside the limits of marketing authorizations and reimbursement by the French universal health insurance system. PMID- 12537260 TI - Diagnostic tools for amyloidosis. AB - Demonstration of amyloid deposits in biopsy specimens is the only means of confirming the diagnosis of amyloidosis. In experienced hands, nonsurgical biopsies of the rectal mucosa or, preferably, of the abdominal fat pad or labial salivary glands provide the diagnosis in 80 to 85% of cases. Immunolabeling studies help to determine the histological type of amyloidosis but are not performed routinely in everyday practice. In patients with a family history of amyloidosis, studies of the genome and amyloid protein can identify the protein variants capable of causing systemic amyloidosis. Once the diagnosis of amyloidosis is established, the extent of systemic involvement with amyloid should be evaluated by performing renal and hepatic function tests, a proteinuria assay, and an echocardiogram. Scintigraphy with radiolabeled serum amyloid P (SAP) component is a rapid and specific investigation that provides a map of the amyloid deposits. Deposits are usually seen in the liver and spleen. SAP component scintigraphy can provide support for the diagnosis of amyloidosis in patients with negative histological studies. Tissue retention of radioactivity predicts survival. PMID- 12537261 TI - Work-related musculoskeletal disorders of the upper limb. AB - Work-related musculoskeletal disorders of the upper limb (WRMSDs-UL) account for over two-thirds of all occupational disorders recognized in France. This broad term encompasses a vast array of disorders whose development is facilitated by environmental factors present at the workplace. Numerous epidemiological studies have established the key role of occupational activities in the genesis of WRMSDs UL. This role is mediated by biomechanical factors (repetitive motion, strenuous effort, extreme joint postures) and/or psychosocial factors. Biological plausibility supports the epidemiological data. The high incidence of WRMSDs-UL indicates a need for greater emphasis on prevention. PMID- 12537262 TI - Rheumatological management of patients with hemophilia. Part II: Muscle hematomas and pseudotumors. AB - Although less common than hemarthroses, muscle hematomas nearly always threaten limb function and can threaten life. Their treatment is not as well standardized as that of hemarthrosis but also rests on clotting factor replacement, prevention of contractures, and rehabilitation. Pseudotumors raise even greater treatment challenges and usually require surgery in adults. Presence of an inhibitor is no longer a contraindication to surgery. In HIV-infected patients, surgery is possible only if the CD4+ count is sufficiently high. PMID- 12537263 TI - Hip involvement in patients with familial Mediterranean fever. A review of ten cases. AB - Hip involvement is uncommon in familial Mediterranean fever (FMF) and can result either from a process specific to this disease or from a coexisting chronic inflammatory joint disease, usually suggestive of ankylosing spondylitis (AS). We report ten cases of FMF with radiologically-documented inflammatory hip disease. Five patients had AS and one had juvenile idiopathic arthritis. There were six men and four women, with a mean age of 34.4 years +/- 17.6 (range, 15-70 years). Onset of the inflammatory hip disease occurred after bouts of acute hip symptoms in one of the patients with isolated FMF and after protracted hip arthritis in another; the two other patients had no history of hip symptoms. The HLA-B27 antigen was looked for in two of the five patients with FMF and AS, with negative results in both; another patient in this subgroup had severe ulcerative colitis. Total hip replacement or replacement of the acetabulum was required in six patients, including two with isolated FMF. Chronic joint disease has been estimated to contribute fewer than 5% of the joint manifestations in FMF. In previous studies, the hips and knees were affected in 75% of patients with chronic joint disease related to FMF. The association of FMF and AS (usually without the HLA-B27 antigen) has been well documented, although the pathogenic mechanisms that link these two conditions remain unknown. PMID- 12537264 TI - Isokinetic evaluation of the knee in patients with rheumatoid arthritis. AB - OBJECTIVES: Rheumatoid arthritis is a chronic inflammatory disease with a clinical picture of arthritis, muscle hypotrophy, loss of range of motion, loss of strength and disability. The objective of this study was to evaluate knees of patients with rheumatoid arthritis using an isokinetic dynamometer. METHODS: Fifty patients with a diagnosis of rheumatoid arthritis and 50 control subjects were evaluated using an isokinetic dynamometer (Cybex 6.000) regarding the following parameters: peak torque, peak torque angle, power, total work, peak torque acceleration time, set total work, torque acceleration energy ('explosion') and endurance. Comparisons between rheumatoid arthritis and the control group, left side versus right side, flexors and extensors and the proportion between flexors and extensors were made. The subjects were also evaluated through the Health Assessment Questionnaire, visual and analogical scale of pain, EPM-ROM and goniometry of the knee. RESULTS: The results showed that patients with rheumatoid arthritis have less strength than the control group (P < 0.05); the extensors are stronger than the flexors (P < 0.05); no significant differences between the right and the left knee for rheumatoid arthritis and the control group were observed and the proportion between flexors and extensors is the same in both groups. We concluded that regarding the isokinetic parameters, the knees of subjects with rheumatoid arthritis are different from normal knees with decreased strength parameters, maintaining the proportion between flexors and extensors with a global loss of strength in the segment, excluding the high speeds in some of the parameters. PMID- 12537265 TI - Sausage-like toe and heel pain: value for diagnosing and evaluating the severity of spondyloarthropathies defined by Amor's criteria. A retrospective study in 161 patients. AB - AIM: To evaluate the frequency of sausage-like toe and talalgia in spondyloarthropathies diagnosed by the 12 items of Amor's criteria; to study the frequency of a diagnosis which would be impossible without these two symptoms; and to study the correlations between Amor's criteria score and the clinical and biological symptoms in the presence of both of these. METHODS: Retrospective study of 161 cases (age, 38.8 +/- 13.1 years; duration, 7.12 +/- 7.3 years; HLA B27, 112). Peripheral involvement is observed in 89 cases (52 cases of pure peripheral form). RESULTS: Eighty cases (group N1) experience talalgia and/or sausage-like toe and 80 do not (group N2). Talalgia concerns 68 patients (42.2%) and sausage-like toe 37 (23.3%); both signs are present in 18. Age, duration, morning stiffness, SR, CRP are similar in both groups. Ankle involvement and oligoarthritis are significantly more frequent in group N1. In 17 cases the presence of talalgia or sausage-like toe are indispensable to the diagnosis (10.6%): 11 undetermined spondyloarthropathies, six psoriatic arthritis. In the N1 group, a positive correlation is present between the score of the 12 items and, respectively, morning stiffness (+ 0.69; P = 0.0001), SR (+ 0.6; P = 0.001 8), CRP (+ 0.59; P = 0.001 8) in patients with sausage-like toe, probably because of the associated oligoarthritis, but neither in patients with talalgia (N1) nor in patients of the group N2. CONCLUSION: Talalgia and sausage-like toe are present respectively in 42.2 and 23.6% of spondyloarthropathies and are indispensable to the diagnosis using Amor's criteria in 10.6% of cases. The sausage-like toe would indicate a more severe spondyloarthopathy because of the synovitis, even though talalgia (pure enthesopathy) does not reflect the inflammatory process. PMID- 12537266 TI - Epidemiology of osteoarthritis and enthesopathies in a European population dating back 7700 years. AB - OBJECTIVES: To investigate the epidemiology of bone changes related to degenerative joint disease and enthesopathies in the earliest Neolithic population identified in Europe (dated back 7700 years), to compare the findings with those in medieval and contemporary populations, and to draw etiological inferences. METHODS: Examination of the skeletal remains of 119 individuals older than 20 years of age at death and buried in two Central European cemeteries dated as far back as 7700 years (5700 BC). RESULTS: The propensity for developing degenerative disease seemed to vary among individuals, as it does today. The hands, shoulder, and hip were common targets of osteoarthritis, as they are today, and the elbow was more likely to be affected in the Neolithic and medieval populations than in contemporary populations. When age of the individuals is taken into account, the prevalence of knee osteoarthritis does not seem to have changed substantially overtime. CONCLUSIONS: These results attest to the major role of microtrauma in osteoarthritis of the upper limb joints. At the other joints, after correction for age, the prevalence of degenerative disease and the combinations of joint involvements have remained remarkably constant over time, suggesting a major role for genetic factors (bone formers) already present in European populations in the early Neolithic period. PMID- 12537267 TI - Computed tomography in low back pain and sciatica. A retrospective study of 132 patients in the Haute-Vienne district of France. AB - AIMS: To evaluate physician compliance with the guidelines of the National Agency for Accreditation and Health Evaluation (ANAES) and the Consensus Conference on the use of medical imagery in low back pain and sciatica. METHODS: We performed a retrospective study of 132 computed tomography scans (CTs) of the lumbar spine performed in one public and one private healthcare facility in the Haute-Vienne district, France. For each patent, the clinical findings, results of other investigations, prescriptions, and procedures reimbursed by the universal health insurance system were recorded. RESULTS: Guidelines on imagery were followed in 2% of patients with chronic nonspecific low back pain. In 72% of patients, CT results had no influence on the subsequent clinical management. The guidelines were followed more often in patients with sciatica: 85% underwent CT more than 4 weeks after the initial painful episode. However, before CT was ordered, only 54% received appropriate initial treatment with analgesics, nonsteroidal anti inflammatory drugs (NSAIDs), and/or muscle relaxants. Among these patients, 25% also received second-line medical therapy consisting of facet joint injection, conventional traction and, after the initial acute phase, physical therapy. In 39% of the sciatica patients, the imaging results had no effect on subsequent management. Among these patients, 12% underwent surgery for disk herniation. CONCLUSIONS: Ten years after the consensus conference and despite the publication of the ANAES guidelines, there is still a wide gap between observed practice and recommendations for optimal management. The consequences of this extend beyond unnecessary expenses for the universal health insurance system to include important deleterious effects on the patients. In particular, prompt appropriate management may help to avoid progression to chronic low back pain and unnecessary imaging studies and surgical procedures, which often have devastating social and occupational consequences. PMID- 12537268 TI - Beneficial effects of information leaflets before spinal steroid injection. AB - How beneficial is the provision of information leaflets to low back pain patients before steroid injection under fluoroscopy? OBJECTIVES: To compare the value of information leaflets with verbal information on steroid injection under fluoroscopy. METHODS: Alternate month design. One hundred and twenty-three low back pain patients hospitalized for steroid injection under fluoroscopy were enrolled in the trial. Fifty-two patients received both written standardized information and non-standardized verbal information (intervention group), seventy one patients received only non-standardized verbal information (control group). Anxiety assessed at baseline evaluation and just before the injection; satisfaction related to the information received assessed on discharge day; knowledge about steroid injection assessed 4 hours and 1 month after the injection. RESULTS: Patients had a high anxiety level at baseline evaluation. Written standardized information did not decrease significantly anxiety (P = 0.068) before the injection, had no effect on pain during the injection, but increased patients' knowledge about the adverse effects on the day of injection and 1 month later (P = 0.040 and P = 0.084 respectively), and improve satisfaction with information received about potential complications of the steroid injections (P = 0.018). CONCLUSIONS: Providing an information leaflet to low back pain patents undergoing steroid injection under fluoroscopy tends to reduce state anxiety, and increases patients' knowledge and satisfaction with information about the risks of the injection. PMID- 12537269 TI - Septic arthritis of the hip caused by Yersinia enterocolitica: a case report. AB - We report a case of bacteriologically documented hip infection caused by Yersinia enterocolitica. A 67-year-old male with a history of valvular disease was admitted for pain and motion range limitation in the left hip with a fever. No organisms were recovered by needle aspiration, but Yersinia enterocolitica grew in joint fluid obtained by surgical arthrotomy. Investigations of the gastrointestinal tract were normal, and there was no evidence of endocarditis. After 6 weeks of appropriate antibiotic therapy and immobilization with transtibial traction, the clinical and laboratory test abnormalities improved. However, the patient died from an intercurrent condition. Y. enterocolitica, a well-known cause of reactive arthritis, can cause septic arthritis. PMID- 12537270 TI - Hip dislocation revealing hip tuberculosis. A case report. AB - Hip dislocation is an uncommon presentation of hip tuberculosis. We report a case in an 18-year-old woman with active hip tuberculosis. An attempt to reduce the dislocation 7 weeks into antituberculous therapy was followed by necrosis of the femoral head. Although severe forms of hip tuberculosis are common in endemic areas, dislocation is exceedingly rare. Capsule laxity and/or synovial hypertrophy probably contribute more to the occurrence of dislocation than does the accumulation of pus. PMID- 12537271 TI - Concomitant compression of median and ulnar nerves in a hemophiliac patient: a case report. AB - A 15-year-old boy, with a diagnosis of hemophilia A, suffered bleeding into his left forearm 5 months before being admitted to our medical center. His neurological examination revealed a pronounced median neuropathy and a minor ulnar neuropathy on the left side. There was marked muscle atrophy on the thenar side and, to a lesser degree, on the hypothenar side and in the forearm. Electromyographic findings demonstrated an evident, nearly complete, sensorimotor axonal loss in the median nerve. Magnetic resonance imaging studies showed atrophy in muscles of the left forearm and median nerve. The patient was diagnosed as having median nerve axonotmesis and ulnar nerve neuropraxia due to compartment syndrome. In hemophiliac patients, frequent single nerve compressions (often involving the femoral nerve) can be seen. However, concomitant median and ulnar nerve injuries with differing severity are rare. PMID- 12537272 TI - Rheumatoid nodulosis: a new case report. PMID- 12537273 TI - Large geodes in rheumatoid arthritis without joint destruction. AB - Although subchondral geodes are a well-known radiological feature of rheumatoid arthritis (RA), large geodes are uncommon. Progressive bone damage with pathological fractures has been reported. We report the case of a 49-year-old man with seropositive RA in whom large, rapidly progressive geodes in the wrists, hands, and feet contrasted with the absence of joint destruction, good functional tolerance, and moderate abnormalities of markers for inflammation. The location and rapid progression of the cyst-like lesions in this patient were highly unusual. PMID- 12537274 TI - Short-term serum C-reactive protein response after shoulder replacement. PMID- 12537276 TI - Introduction: health expectations for older women: international perspectives. PMID- 12537275 TI - A primary non-Hodgkin's lymphoma of bone in Tunisia. A retrospective study of 15 cases. PMID- 12537277 TI - Patterns of active life among older women: differences within and between groups. AB - This study examines the distribution of total, unimpaired, and impaired life for several groups of older women defined by race, education, and marital history. Using data from the 1984-1990 Longitudinal Study of Aging, we model transitions among functional statuses using discrete-time Markov chains, and use microsimulation to produce summary indices of active life. Remaining years of life and the proportion of remaining years with disability vary substantially, both within each group of women studied and between pairs of groups. Of all groups studied, never-married, more-educated white women live the longest, healthiest lives. Ever-married nonwhite women with low education have the shortest life expectancy, and experience the most disability. Our findings show that life expectancy is an incomplete indicator of the time women, in particular sub-groups, can expect to live with and without impairment. These findings highlight the heterogeneity of disability processes and life expectancy for older women. PMID- 12537278 TI - Health profiles and profile-specific health expectancies of older women and men: The Netherlands. AB - This study focuses on gender differences in health profiles, and examines which health profiles drive gender differences in remaining life expectancy in women and men aged 65 and over in The Netherlands. Data from the first two cycles of the Longitudinal Aging Study Amsterdam (n = 2,141 and 1,659, respectively) were used to calculate health profiles for individuals of 65-85 years. For both women and men, six profiles were found: I. cancer; II. "other" chronic diseases; III. cognitive impairment; IV. frailty or multimorbidity; V. cardiovascular diseases; and VI. good health. The further characterization of these types showed some gender differences. Remaining life expectancy for women was greater than for men in each health profile. A decomposition into health expectancies showed that both women and men could expect to live about 5 years in good health from age 66. The greatest gender differences in years spent with health problems were found for profile IV and for profile III. Their greater number of years spent in these health states have direct consequences for the type and cost of care women need. PMID- 12537279 TI - Life with and without disease: women experience more of both. AB - This paper examines gender differences in life with and without six major diseases, including both mortal and morbid conditions. Disease prevalence and health behavior data are from the 1993-1995 National Health Interview Surveys for the United States. Vital registration data are the source of mortality rates used in computing life expectancy. The Sullivan method is used to estimate life lived with and without disease and risky behavior for men and women at various ages. Women live more years with each of the diseases examined, and, for arthritis, the extended years with disease are greatest. Women also live more years than men free of each of these diseases with the exception of arthritis. Gender differences in life without two health-risk behaviors are also discussed. Men spend more years of their lives overweight and have fewer years during which they see a doctor. PMID- 12537280 TI - Gender differences in disability-free life expectancy for selected risk factors and chronic conditions in Canada. AB - This article shows how mortality and morbidity patterns differ for women and men 45 years of age and older. The impact on disability-free life expectancy was calculated for selected risk factors and chronic conditions: low income, low education, abnormal body mass index, lack of physical activity, smoking, cancer, diabetes, and arthritis. For each factor, the expected number of years free of disability was calculated for men and women using multi-state life tables. In terms of disability-free life expectancy, the greatest impacts on affected women were for diabetes (14.1 years), arthritis (8.8 years), and physical inactivity (6.0 years), while for affected men, the greatest impacts were for diabetes (10.5 years), smoking (6.9 years), arthritis (6.5 years), and cancer (6.4 years). The implications of these results are discussed from the perspective of developing programs designed to improve population health status. PMID- 12537281 TI - Gender differences in life expectancy free of impairment at older ages. AB - This article uses data from the United Kingdom Medical Research Council Cognitive Function and Ageing study (MRC CFAS) to analyze morbidity associated with three areas of impairment. We use cognitive status, functional status, and physical illness to examine differences in the proportion of time that older women and men will spend with co-morbidity. We also analyze differences among various impairments, and investigate the relationship between missing data and sex. Women have a larger burden of impairment than men, and, by including cognitive impairment together with functional impairment, a very large impairment burden is highlighted at all ages. Policy implications of the findings from the perspective of older women in the United Kingdom are discussed. PMID- 12537282 TI - Global patterns of healthy life expectancy for older women. AB - This paper focuses on patterns of healthy life expectancy for older women around the globe in the year 2000, and on the determinants of differences in disease and injury for older ages. Our study uses data from the World Health Organization for women and men in 191 countries. These data include a summary measure of population health, healthy life expectancy (HALE), which measures the number of years of life expected to be lived in good health, and a complementary measure of the loss of health (disability-adjusted life years or DALYs) due to a comprehensive set of disease and injury causes. We examine two topics in detail: (1) cross-national patterns of female-male differences in healthy life expectancy at age 60; and (2) identification of the major injury and disability causes of disability in women at older ages. Globally, the male-female gap is lower for HALE than for total life expectancy. The sex gap is highest for Russia (10.0 years) and lowest in North Africa and the Middle East, where males and females have similar levels of healthy life expectancy, and in some cases, females have lower levels of healthy life expectancy. We discuss the implications of the findings for international health policy. PMID- 12537283 TI - European perspectives on healthy aging in women. AB - Using data from the 1994 European Community Household Panel, we compare active life expectancy differentials at age 65 years between women and men in 12 European countries. We seek to explain the extent to which differences are a reflection of gender differentials in life expectancy at 65 years or reflect differences in active life expectancy earlier in life. Considerable variation in the gender differentials in both total and active life expectancies at age 65 years exist within Europe, with some countries experiencing 20% lower life expectancy at age 65 years for men compared to women. Some evidence was found to suggest that gender differentials in active life expectancy may continue from younger ages through to later life. PMID- 12537284 TI - Health expectancies in Japan: gender differences and policy implications for women. AB - Based on prospective observation of elderly people in the community in Japan, we compared the time-course of development and progression of physical disability between women and men. Men experienced disability at a younger age and at a faster rate than did women. The duration of time spent with disability in women was twice as long as in men. Consequently, women consume about two-thirds of the total resources of formal caregiving services in Japan. Women in Japan are increasingly educated, postponing marriage to higher ages, and less likely to care for parents in the home. Given these changes in family structure and social norms, the capacity for informal family caregiving has decreased dramatically. A recently enacted national long-term care insurance system may further change the picture of caregiving. PMID- 12537285 TI - Disability among older women and men in Fiji: concerns for the future. AB - This study examines the composition of elderly population at risk of disability and speculates the impact of disability on the quality of their lives and their longevity. Using census and survey data collected in Fiji, life table estimates of unimpaired life expectancy across time are presented for older people and the potential costs of disability, in terms of productive years of life lost. From a planning perspective, the study discusses medical and support services that may be needed to support older individuals in Fiji. The study also describes policy implications of the findings, focusing on the older women, and considers the implications for older women of other developing countries. PMID- 12537286 TI - Recent perspectives on active life expectancy for older women. AB - This article provides a critical review of recent active life expectancy literature, describing trends of special interest to women. We review findings from leading perspectives used to study life expectancy and active life expectancy, including gender, racial and socioeconomic differences, disease specific effects, and biodemography. We examine three competing theories of population health that frame active life expectancy research-compression of morbidity, expansion of morbidity, and dynamic equilibrium-concluding there is support for both the compression of morbidity and dynamic equilibrium theories. Policy implications for women include a greater understanding of the role of education and racial and ethnic diversity in active life trends, and an increased public policy emphasis on prevention and treatment of chronic disease, together with adoption of more healthy lifestyles. PMID- 12537287 TI - Provisional surveillance summary of the West Nile virus epidemic--United States, January-November 2002. AB - This report presents provisional summary surveillance data about West Nile virus (WNV) activity that were reported to CDC during January 1-November 30, 2002, through the ArboNET surveillance network. In 2002, the reported numbers of human and animal infections increased, and the geographic range of WNV activity expanded substantially. These data underscore the need for intensive surveillance to detect and quantify WNV activity in areas where humans are at risk, public education to teach persons how to prevent mosquito bites, and sustained and integrated mosquito-control activities. PMID- 12537288 TI - Laboratory-acquired West Nile virus infections--United States, 2002. AB - West Nile virus (WNV), a mosquito-borne flavivirus introduced recently to North America, is a human, equine, and avian neuropathogen. The majority of human infections with WNV are mosquito-borne; however, laboratory-acquired infections with WNV and other arboviruses also occur. This report summarizes two recent cases of WNV infection in laboratory workers without other known risk factors who acquired infection through percutaneous inoculation. Laboratory workers handling fluids or tissues known or suspected to be WNV-infected should minimize their risk for exposure and should report injuries and illnesses of suspected occupational origin to their supervisor. PMID- 12537289 TI - Intrauterine West Nile virus infection--New York, 2002. AB - West Nile virus (WNV), a mosquito-borne flavivirus and human neuropathogen, is epidemic in the United States. In 2002, newly recognized mechanisms of person-to person WNV transmission were described, including possible transmission from mother to infant through breast milk. WNV has not been previously associated with intrauterine infection or adverse birth outcomes. This report describes a case of transplacental WNV transmission. Pregnant women should take precautions to reduce their risk for WNV or other arboviral infection and should undergo diagnostic testing when clinically appropriate. PMID- 12537290 TI - Enzyme polymorphisms in Rhizoglyphus robini and R. echinopus and their application in paternity analysis. AB - Cellulose acetate electrophoresis (CAE) was used to investigate enzyme polymorphism in two congeneric species of Acaridae, Rhizoglyphus robini and R. echinopus. Using homogenates of individual mites, 27 enzymes were examined in two buffers. Five enzymes showed interspecific polymorphism, 15 exhibited intraspecific variation in R. robini and 12 in R. echinopus. Polymorphic PGDH was used to analyse paternity in progeny of R. robini females that had mated with two males. The second male sired on average 67% of progeny. Enzyme polymorphism can be used to study poorly known aspects of acarid mites biology, such as their mating systems and population structure under natural conditions and in stored foods. PMID- 12537291 TI - Diel periodicity of emergence of adult citrus rust mites in central Florida. AB - Emergence of adult citrus rust mites, Phyllocoptruta oleivora (Ashmead) from the imagochrysalis occurred with a diel periodicity that peaked between mid morning and early afternoon. The pattern of emergence did not differ for mites reared from eggs held under different photoperiod regimes. The transition of nymphs to the imagochrysalis stage also showed a diel periodicity, occurring in the evening. The average duration of the imagochrysalis stage at 28 degrees C was 14 15 h. Wetting by rain or dew was simulated by immersing fruit in water each night, but the transition to the imagochrysalis stage or the subsequent emergence pattern of adults was not affected. Adult mites reared from eggs laid during two, consecutive 12-h periods emerged at the same time of day, but the peaks of emergence were separated by 24 h. Eclosion of larvae from eggs laid during two, consecutive 12-h periods did not show a diel periodicity. Eggs continuously immersed in water during their development did not hatch, whereas there was a small, but significant, reduction in the viability of eggs immersed in water each night. These results are discussed in relation to external cues that may entrain the diel patterns of development and adult emergence, and to the adaptive responses of citrus rust mite to cyclic fluctuations of key environmental variables in central Florida. PMID- 12537292 TI - Morphological variation in different host populations of Abacarus hystrix (Acari: Prostigmata: Eriophyoidea). AB - The present systematic classification of Eriophyoidea is widely recognised as artificial and not reflecting plant-herbivore linkage. Quantitative description of host-related morphological variation can provide the basic information needed to improve the eriophyoid taxonomic system and enhance our understanding of mechanisms generating this variation. The purpose of this study was to investigate quantitative morphological traits of populations of cereal rust mite Abacarus hystrix (Nalepa 1896) living on different hosts. Three populations of A. hystrix collected from different grasses (Lolium perenne, Bromus inermis and Elytrigia repens) were examined morphologically. MANOVA analysis revealed significant differences in vectors of means among the three populations. Discriminant analysis yielded 11 traits that significantly differentiate the three populations. Analysis of canonical loadings showed that traits, which best discriminate the populations living on different hosts, are: body elongation, length of setae and overall body size. Host-dependent morphological variation is interpreted in terms of adaptation to specific environmental conditions created by the host. Hypotheses on the sources of this variation are discussed. PMID- 12537293 TI - Susceptibility of cypress seedlings to the eriophyoid mite Trisetacus juniperinus. AB - In Italian nurseries and young groves of evergreen cypress (Cupressus sempervirens L.), the eriophyoid mite Trisetacus juniperinus (Nal.) is considered a very serious pest. A rating system of damage symptoms was developed to investigate the susceptibility of different cypress seedling families to the mite. Based on this system, the seedlings were evaluated for three years in the nursery and in two field locations after transplanting. Data obtained in the nursery allowed the cypress families to be allocated to at least two different levels of susceptibility. These levels of susceptibility were generally also maintained in the field. However, the environmental conditions of the two transplanting localities significantly affected the susceptibility of each family. In all families, scores for each of the damage categories were strongly correlated positively to each other and negatively with the average increment in the height of plant over the duration of the field experiment. Assessment of the intensity of the symptoms peculiar to damage category A (buds enlarged, deformed, russet and/or branch apex folded) was sufficient to give the same susceptibility evaluation as if data for all damage categories were used. The evaluation of susceptibility on the basis of injury pattern may return very useful information for selection and certification of families of known susceptibility to eriophyoid mites. PMID- 12537294 TI - Demographic traits of Eotetranychus pruni from Belgian and Tunisian orchards, in comparison with Tetranychus urticae. AB - Eotetranychus pruni was recorded in Belgian and Tunisian apple orchards for the first time. The aim of this study was to compare a Belgian (Vieusart) and a Tunisian strain (Grombalia), with respect to their life history and demographic traits. Furthermore, a Tunisian strain of Tetranychus urticae (Sousse) was also added to the study, in order to evaluate the potential status of the Tunisian strain of E. pruni. Experiments were carried out in the laboratory (24 degrees C) on apple leaf discs. Significant differences were noticed between the two species, with respect to development time (males), fecundity and fertility (egg hatching), but no differences could be detected between the two strains of E. pruni. Life tables were set up to describe the population statistics. The calculated intrinsic rates of natural increase (rm) appear to be rather low per se and do not correspond to those of typical "r strategic pests". The Bootstrap procedure allows us to estimate the confidence interval for these rm values. By using this method, the two Tunisian strains (T. urticae and E. pruni) appear to be quite similar, and contrast with the Belgian strain (E. pruni). The potential pest status of the Tunisian strain of E. pruni is discussed. PMID- 12537295 TI - Pyrethroid-Acarophenax lacunatus interaction in suppressing the beetle Rhyzopertha dominica on stored wheat. AB - Any biocontrol agent used against stored grain insects will have to interact with insecticides used as grain protectants due to their heavy use especially in tropical areas. Therefore, the interaction between the pyrethroid insecticides bifenthrin and deltamethrin and Acarophenax lacunatus (Cross & Krantz) (Prostigmata: Acarophenacidae), an egg parasite of the stored cereal pest Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae), was assessed in a range of mite densities and insecticide doses for each compound. Despite the mite species presence on all insecticide doses, the lowest instantaneous rate of increase of A. lacanatus was recorded with the highest doses of insecticides. The presence of the biological control agent at all doses of both insecticides suggests its tolerance to these compounds. A. lacunatus showed higher parasitism rate with bifenthrin than with deltamethrin indicating a higher selectivity of the first compound to the biocontrol agent. PMID- 12537296 TI - Resistance to hexythiazox in Brevipalpus phoenicis (Acari: Tenuipalpidae) from Brazilian citrus. AB - The objective of this study was to collect baseline information for implementing an acaricide resistance management program of Brevipalpus phoenicis (Geijskes) to hexythiazox in Brazilian citrus groves. The egg susceptibility of B. phoenicis to hexythiazox was measured by a direct contact bioassay. The estimated LC50 for the S strain was 0.89 mg hexythiazox L(-1) of water (95% FL 0.75-1.03). After hexythiazox selection of a field-collected population associated with intense hexythiazox use, a resistance ratio greater than 10,000-fold was detected. Results from a survey revealed a great variability in the frequency of resistance in populations of B. phoenicis collected from citrus groves located in the State of Sao Paulo. No relationship was observed between the intensity of hexythiazox use and the frequency of resistance. Studies on dynamics of resistance showed that the resistance of B. phoenicis to hexythiazox is stable under laboratory conditions. Therefore, there is an urgent need to implement resistance management of B. phoenicis to hexythiazox in order to prolong its effective use in Brazilian citrus groves. PMID- 12537297 TI - The stability of tebufenpyrad resistance in two-spotted spider mite (Acari: Tetranychidae) under laboratory conditions. AB - The successful use of chemical rotations to manage insecticide resistance requires reversion between alternate chemical applications. We tested a tebufenpyrad resistant population of Tetranychus urticae Koch after some 55 months laboratory culture without pesticide selection and found LC50 level resistance had dropped from 63.29- to 2.41-fold. However, the population was still heterogeneous with LC99 level resistance at 38.03-fold. It is likely that a lack of reversion contributed directly to the initial tebufenpyrad control failure. PMID- 12537298 TI - Parasitism of lizards by immature stages of the blacklegged tick, Ixodes scapularis (Acari, Ixodidae). AB - From 1982-1985 and 1993-1999, a total of 309 individual reptiles, mostly lizards and snakes, belonging to 12 species (American alligator, six lizard species, five snake species) was captured on St. Catherine's Island, Liberty County, Georgia, USA, and examined for ticks. Three lizard species, the broad-headed skink Eumeces laticeps, southeastern 5-lined skink Eumeces inexpectatus, and eastern glass lizard Ophisaurus ventralis, were severely infested with larvae and nymphs of the blacklegged tick, Ixodes scapularis. Ticks were not found on any of the other reptile species. Overall, 80% of 65 E. inexpectatus examined were parasitized by a mean intensity of 21.5 larvae and 88% were parasitized by a mean intensity of 4.8 nymphs. Corresponding figures for E. laticeps (n=56) were 93% and 51.3 for larvae and 89% and 7.4 for nymphs, and for O. ventralis (n=3) were 67% and 22.5 for larvae and 100% and 21.3 for nymphs. Larvae and nymphs attached along the lateral grooves of O. ventralis. Nymphs attached mainly behind the ears and in the foreleg axillae whereas larvae mainly attached to these sites and on the hindlegs in Eumeces spp. Seasonally, both larvae and nymphs were recorded on lizards from April through October. A unimodal larval peak was recorded in May or June. Seasonal data for nymphs did not reveal any distinct peaks but small bimodal peaks in mean intensities may have occurred (one in early summer, the other in late summer) suggesting that some ticks complete their life cycle in one year, and others in two years, on St. Catherine's Island. Potential epidemiological consequences of these findings with respect to Lyme disease in the southeastern United States are briefly addressed. PMID- 12537299 TI - Intracellular Ca++/Mg++ homeostasis during postnatal growth of experimental rats. Multiple time-point study. AB - In most tissues, various cell membrane ion transporting systems are not fully developed and/or maximally active at the prenatal and early postnatal stage. Their progressive development and expression are a function of growth and maturity. We performed a multiple time-point study, in order to investigate the ability of a variety of tissues to maintain appropriate Ca++ and Mg++ homeostasis at different stages of postnatal development. Total intracellular Ca++ in one week-old rat liver, brain and spinal cord tissues was significantly elevated, compared to mature animals. It increased further through the first three weeks of gestation. Intracellular Ca++ gradually and significantly declined in adult and mature animal groups. Alterations in total intracellular Mg++ of the same tissue samples, although not so profound, paralleled changes in total intracellular Ca++. We conclude that a developmental switch in intracellular Ca++ and Mg++ homeostasis occurs one to three weeks following birth. It might be related to the incomplete development of Ca++ and Mg++ transmembrane transporting systems, previously reported as being only partially expressed at the early postnatal stage. These developmental alterations in total intracellular Ca++ and Mg++ content might serve as a regulatory mechanism, adjusting cell activities to the physiological requirements of the growing and maturing animal. PMID- 12537300 TI - Assessing the effect of litter size on growth pattern and homeothermy acquisition in the Pampas mice Akodon azarae (Rodentia, Muridae). AB - We evaluated in pups of Akodon azarae both the growth pattern from birth to 48 days of age and the effect of litter size on growth pattern and homeothermy acquisition from birth to weaning age. Individual pups gained weight as expected by a Gompertz growth pattern. Until weaning, litter size affected both the slope of the relationship between body temperature and age and the rate of growth of pups. Pups from small litters increased both body temperature and weight until body temperature and body weight of adults at higher rates than those from large litters. PMID- 12537301 TI - Growth and development of male "little" mice assessed with Parks' theory of feeding and growth. AB - This work was designed to characterize the appetite kinetics and growth of male C57BL/6J (lit) mice. Those variables were assessed with Parks' function of ad libitum feeding and growth. Heterozygous mice (lit/+) attained their mature weight at 12-15 weeks of age, peak growth rate (3.5 g/week) at 5 weeks and displayed the normal decay of food conversion efficiency as a function of age. The homozygous genotype has a chronic defect in the synthesis and secretion of growth hormone (GH). Homozygous mice could not be assessed with Park's function. From the 4th to the 15th week of age, body weight increased linearly and exhibited constant food conversion efficiency. Food intake of both genotypes was commensurate with their body weights. Lit/lit mice became progressively obese. At 40 weeks of age, body fat of lit/lit mice was fivefold that of lit/+ and their body weight was similar to their heterozygous controls. The chronic deficiency of growth hormone produced a lower bone mass (compared to heterozygous controls). Bone mass of both genotypes attained maturity at 12-15 weeks with a maximum growth rate at 5 weeks. Body weight and bone mass grow harmoniously in lit/+ but not in lit/lit mice. PMID- 12537302 TI - Differences in muscle fiber growth in slow-twitch muscles of the forelimb and hindlimb of the rat: role of the pituitary and food intake. AB - This investigation tested the hypothesis that differences in the growth of fore- and hindlimb muscles in the rat are regulated by the pituitary and food intake. Using morphometric techniques, the growth of muscle fibers was compared in two slow-twitch muscles, the flexor carpi ulnaris (FCU) of the forelimb, and the soleus of the hindlimb, in male Wistar rats fed ad libitum, food restricted (FR) or hypophysectomized (hypox) from age 60 days. Growth was defined as an increase in fiber diameter and/or type 1 fiber percentage. The soleus had larger diameter fibers than the FCU in controls and FR, but not hypox rats. The growth in diameter, between 60 and 180 days, of both types 1 and 2 fibers in the soleus and type 2 fibers in the FCU was inhibited by hypox and, to a lesser extent, FR. Neither type 1 fiber diameter nor percentage of type 1 fibers in the FCU increased with age nor was it affected by hypox or FR. The percentage of type 1 fibers was higher in the soleus than the FCU and was further increased in the soleus of hypox rats. Food restriction produced a smaller rise than hypox in type 1 fiber percentage in the soleus. Thus, differences in fore- and hindlimb muscle fiber growth are modulated by pituitary hormones and, to a lesser extent, by food intake. PMID- 12537303 TI - Intracerebral sources of human auditory steady-state responses. AB - The objective of this study was to localize the intracerebral generators for auditory steady-state responses. The stimulus was a continuous 1000-Hz tone presented to the right or left ear at 70 dBSPL. The tone was sinusoidally amplitude-modulated to a depth of 100% at 12, 39, or 88 Hz. Responses recorded from 47 electrodes on the head were transformed into the frequency domain. Brain electrical source analysis treated the real and imaginary components of the response in the frequency domain as independent samples. The latency of the source activity was estimated from the phase of the source waveform. The main source model contained a midline brainstem generator with two components (one vertical and lateral) and cortical sources in the left and right supratemporal plane, each containing tangential and radial components. At 88 Hz, the largest activity occurred in the brainstem and subsequent cortical activity was minor. At 39 Hz, the initial brainstem component remained and significant activity also occurred in the cortical sources, with the tangential activity being larger than the radial. The 12-Hz responses were small, but suggested combined activation of both brainstem and cortical sources. Estimated latencies decreased for all source waveforms as modulation frequency increased and were shorter for the brainstem compared to cortical sources. These results suggest that the whole auditory nervous system is activated by modulated tones, with the cortex being more sensitive to slower modulation frequencies. PMID- 12537304 TI - Task-specific expectation is revealed in scalp-recorded slow potentials. AB - The contingent negative variation (CNV) is known as an electrical manifestation of expectancy, readiness and attention. The purpose of this study was to evaluate whether slow potentials recorded from the human scalp during the expectation to perform different tasks or during the expectation of the second stimulus in the pair demonstrate specificity regarding the type of visual stimuli. Participants were nineteen healthy adults. Each trial began with one of the two cues (S1) followed by consecutive pictures (S2 and S3). Each picture of this pair was a photograph of a familiar face with superimposed abstract dot pattern. One cue directed attention to compare faces and another to compare patterns. EEG was recorded with a 128-channel net. Slow potentials following S1 and S2 presentation (SP1 and SP2) are reported here for Face and Pattern tasks. The duration of each segment of the trial was 1500 ms from the stimulus onset, and here the last 1000 ms was analyzed. Statistical comparisons of amplitudes and current density mapping were performed. Within the same task, SP1-SP2 differences were complex, implying multiple dissimilarities in the processes during the two expectation periods. Slow potentials in SP1 differed between the tasks mostly in the anterior recording sites in 700-1500 ms. These latencies corresponded to the late CNV (or stimulus preceding negativity, SPN) showing task-specificity during SP1. For SP2, the differences were located centrally in 500-900 ms, the period that reflects the superimposition of processes generating the early CNV and the late positive complex. Task-specific differences appeared only in SP1. PMID- 12537305 TI - Reliability and validity of neuroelectric source imaging in primary somatosensory cortex of human upper limb amputees. AB - The present study investigated the test-retest reliability of EEG source localizations of somatosensory evoked potentials (SEPs) in human upper limb amputees over a long time frame (several months) and examined the validity of source reconstruction. In two sessions spaced several months apart five unilateral upper limb amputees were stimulated at the first and fifth digit of the intact hand and at the left and right lower corner of the mouth. To examine the validity of the results of the neuroelectric source reconstruction a comparison with neuromagnetic source localization was performed for two subjects. The source localizations of the SEP components were found to be highly reproducible: the mean standard deviation of the dipole locations was 8.80 mm in the x-, 7.00 mm in the y- and 4.15 mm in the z-direction. The match of the comparison of EEG and MEG data was in the range of one centimeter. These results support the use of multi-electrode EEG recordings combined with MRI as an adequate method for the investigation of the functional organization of the somatosensory cortex in upper limb amputees and suggest high stability of cortical reorganization in these subjects. PMID- 12537306 TI - Detection of non-linearity in schizophrenic patients using magnetoencephalography. AB - OBJECTIVE: The aim of this study is to investigate the presence of any non linearity in the magnetoencephalographic recordings (MEG) from the temporal lobe of schizophrenic patients in comparison with controls, in order to find the differences underlying the brain waves. We calculated the correlation dimension, which is a measure of the complexity of the dynamic system, as well as the first Lyapunov exponent that indicates the system's unpredictability. METHODS: The schizophrenic group consisted of 3 men and 7 women aged 23-32 years (mean 27.2, SD=3.5) and the control group of 3 men and 6 women aged 26-35 years (mean 31.6 +/ 4.1). There were no significant differences between the two groups as far as age and sex were concerned. None of them received any medication. RESULTS: The analysis of the MEG in the schizophrenic group showed lower dimension complexity and moreover the first Lyapunov exponent presented lower values compared with the corresponding ones in the control group, which means lower information processing. CONCLUSION: EEG findings as determined by MEG and non-linear analysis may offer important perspectives to better understand brain function in schizophrenia. PMID- 12537307 TI - Contact heat evoked potentials to painful and non-painful stimuli: effect of attention towards stimulus properties. AB - The study aim was to evaluate the effect of different attentional tasks on the amplitudes and latencies of painful and non-painful contact heat evoked potentials (CHEPs). CHEPs were recorded in 12 healthy subjects during two experimental conditions, in which attention was oriented towards the intensity and the distress caused by the stimuli and were compared with CHEPs recorded during a neutral condition. The painful heat stimulation produced a negative potential at Cz vertex with a latency around 540 ms (Cz/N540), a positive peak at Cz electrode around 730 ms (Cz/P730) and, lastly, a positive peak around 1000 ms (Pz/P1000) in the Pz traces. The Cz/P730 wave was significantly higher in amplitude only during the painful stimulation and is probably related to coding the nociceptive activity. Varying the attentional target towards different properties of the stimulus did not cause any significant change in CHEP responses amplitude and latencies compared with the neutral condition. Our results suggest that CHEPs represent a reliable functional measure of the nociceptive pathways and that they are generated by the activation of different cerebral areas involved in pain processing. The high activation level of each of these area or their spatial neighbouring might explain the strong similarity of CHEP components recorded during different attentional manipulations. PMID- 12537308 TI - Single-trial classification of elementary comparison processes on the basis of instantaneous EEG and MEG coherences. AB - The present study is focused on the evidence of possible single-trial EEG/MEG analysis of information processing. The discrimination between thinking modalities of concept activation and pattern comparison for single tasks of elementary comparison procedures is investigated. A neural network classifier with backpropagation learning algorithm is used. The input vector is constructed by parameters of instantaneous coherence (13-20 Hz) between several channel pairs of the EEG and/or of the MEG. Thereby, the strength of synchronization and the time location of synchronization phenomena are taken into consideration. The combination of EEG and MEG coherence parameters led to a classification accuracy of 85-94% for single subjects. Generally, results reached by neural network classifier show a better generalization than linear discriminant analysis. PMID- 12537309 TI - THG113: a novel selective FP antagonist that delays preterm labor. AB - PGF2alpha is an important smooth muscle contractile agent that exerts significant effects on myometrium and is implicated in labor. THG113 was recently identified as a PGF2alpha receptor (FP) antagonist. We characterized the specificity and selectivity of THG113, tested its effects on PGF2alpha-induced smooth muscle contraction, and assessed its efficacy in a model of endotoxin (LPS)-induced preterm labor. [125I]THG113 bound specifically to FP-expressing but not to native (not expressing FP) HEK293 cells. In FP-expressing HEK293 cells, THG113 markedly reduced PGF2alpha-elicited phosphoinositide hydrolysis (IC50 27 nM). Similarly, PGF2alpha-evoked microvascular (retinal) contraction was noncompetitively blocked (by > 90%) by THG113. In contrast, contraction to agonists of homologous prostanoid receptors EP1 and TP (17-phenyl-trinor PGE2 and U46619) was unaffected (< 1%) by high concentrations of THG113 (100 micromol/L); THG113 (100 micromol/L) also did not affect contraction to numerous other agents including platelet activating factor, endothelin, and angiotensin II. Force and duration of PGF2alpha-evoked contractions of myometrial strips of pig (non-pregnant, luteal phase) and mouse (immediately postpartum) were markedly reduced by THG113. In an endotoxin-induced preterm mouse model, lipopolysaccharide (50 microg intraperitioneal) injection at 16 days' gestation resulted in 100% delivery within 15 h; in contrast, 70% of those treated with THG113 (1 mg/day) delivered > 24 h later (at 18 days' gestation; term: 19 days). In addition, in mice injected with lipopolysaccharide and treated 6 h later with THG113 (0.1 mg bolus followed by 1 mg/day) 40% delivered > 48 h later. Fetuses of pregnant mice treated with THG113 were born alive, had higher birth weights (1.6 +/- 0.1 v 1.4 +/- 0.05 g), and appeared healthy. This study describes an effective and selective noncompetitive FP antagonist, THG113, which significantly delays preterm delivery; this provides the basis for future investigations for its use in tocolysis. PMID- 12537310 TI - The adverse renal effects of prostaglandin-synthesis inhibitors in the newborn rabbit. AB - Nonsteroidal anti-inflammatory drugs are frequently used during pregnancy (premature labor, polyhydramnios) and the immediate postnatal period (closure of patent ductus arteriosus). This article evaluates the renal effect of 3 nonspecific COX inhibitors (aspirin, indomethacin, and ibuprofen) in newborn rabbits. Five groups of anesthetized, ventilated, normoxemic 6-day-old rabbits (n = 52) were administered intravenous aspirin (40 mg/kg), indomethacin (2 mg/kg), and ibuprofen (0.02, 0.2, 2.0 mg/kg, respectively). Renal function and hemodynamics as assessed by inulin and para-aminohippuric acid clearances were measured before and in the hour after drug administration. In all groups of animals, the nonselective COX inhibitors induced an increase in renal vascular resistance and a consequent decrease in glomerular filtration rate and renal blood flow. Urine flow rate decreased significantly in all groups, except in the group receiving the lowest dose of ibuprofen. In newborn rabbits, aspirin, indomethacin, and ibuprofen induced intense renal vasoconstriction, which resulted in impaired renal function. This observation illustrates the major renal protective role played by the vasodilatory prostaglandins during the neonatal period, when the kidney is perfused at very low perfusion pressure. We conclude that all COX inhibitors should be administered with the same caution to the preterm neonate. PMID- 12537311 TI - Hyperoxemia caused by resuscitation with pure oxygen may alter intracellular redox status by increasing oxidized glutathione in asphyxiated newly born infants. AB - In a prospective, randomized, blinded trial we have studied the effects of resuscitation upon oxygenation in a group of asphyxiated newly born infants receiving room air or 100% oxygen as the gas source. During the acute phase of asphyxia and until the resuscitation procedure concluded, we determined serial blood gases as well as reduced and oxidized glutathione, enzymes involved in the glutathione redox cycle, and antioxidant enzyme activities. The use of 100% oxygen caused a remarkable increase of partial pressures of oxygen in arterial blood, with values that were frequently above physiological levels (> 100 mm Hg). In addition, we have found a significant correlation between hyperoxemia and the intra-erythrocyte GSSG (oxidized glutathione) concentration. We hypothesize that hyperoxemia may be 1 of the triggering factors responsible for an increased oxidation of GSH (reduced glutathione). Moreover, an increased antioxidant enzyme activity, which reflects an oxidative stress, indicates that the antioxidant capacity of the newly born infant may have been surpassed. PMID- 12537312 TI - The effect of blood transfusion on the hemoglobin oxygen dissociation curve of very early preterm infants during the first week of life. AB - A study was conducted during the first week of life to determine the changes in P50 (PO2 required to achieve a saturation of 50% at pH 7.4 and 37 degrees C) and the proportions of fetal hemoglobin (HbF) and adult hemoglobin (HbA) prior to and after transfusion in very early preterm infants. Eleven infants with a gestational age < or = 27 weeks have been included in study. The hemoglobin dissociation curve and the P50 was determined by Hemox-analyser. Liquid chromatography was also performed to determine the proportions of HbF and HbA. The mean gestational age of the 11 infants was 25.1 weeks (+/- 1 weeks) and their mean birth weight was 736 g (+/- 125 g). They received 26.9 mL/kg of packed red cells. The mean P50 prior and after transfusion was 18.5 +/- 0.8 and 21.0 +/- 1 mm Hg (P = .0003) while the mean percentage of HbF was 92.9 +/- 1.1 and 42.6 +/- 5.7%, respectively. The data of this study show a decrease of hemoglobin oxygen affinity as a result of blood transfusion in very early preterm infants prone to O2 toxicity. The shift in HbO2 curve after transfusion should be taken into consideration when oxygen therapy is being regulated for these infants. PMID- 12537313 TI - Combined effects of bilirubin and hypoxia on cultured neurons from the developing rat forebrain. AB - Hyperbilirubinemia and hypoxia are common causes of brain injury in the newborn. To determine the effects of free bilirubin associated with transient hypoxia on developing rat neurons, the cells were exposed to bilirubin (0.25 to 5 micromol/L) and/or to hypoxia for 3 or 6 hours (95% N2-5% CO2). Glutamate receptor antagonists were added to some cultures. Cell death characteristics, energy metabolism, and protein synthesis were analyzed for 96 hours. Bilirubin increased apoptotic cell death. When associated with hypoxia, the neuronal loss was worsened. Bilirubin reduced energy metabolism, whereas a 6-hour exposure to hypoxia increased it for at least 24 hours, with no influence of additional bilirubin. Bilirubin with or without hypoxia induced 2 increases in protein synthesis, at 1 and 72 hours. In this model, bilirubin may promote programmed neuronal death. When bilirubin is associated with hypoxia, the deleterious effects are enhanced. The suppression of bilirubin induced neuronal damage by the NMDA (N-methyl-D-aspartate) receptor antagonist MK801 suggests the involvement of glutamate. PMID- 12537314 TI - The flow-pressure plot: a new look on the patient-ventilator interaction in neonatal care. AB - Most modern neonatal ventilators have now a built-in flow sensor and, as a spin off of their mechanical action, provide some information about lung function characteristics as compliance and resistance after computation of the flow and pressure signals. Additionally, respiratory graphics as volume-pressure and flow volume plots can be displayed. In clinical practice, however, they are rarely used to refine the ventilator setting. A nonconventional flow-pressure plot is presented in this article, constructed from the volume or flow, and pressure outputs of a Babylog 8000 (Drager, Lubeck, Germany). This flow-pressure diagram appears to be useful in the real-time computation of respiratory mechanics based on the Rahn's law of respiratory motion. Its major advantage, however, is the easy pattern recognition of subtle changes in infant-ventilator interaction, ie, excessive triggering, fighting against the ventilator, augmented breath, tube subobstruction. It may be useful to add the flow-pressure plot to the classical respiratory graphics allowing to monitor mechanical ventilation more accurately and to fine tune the ventilator setting accordingly. PMID- 12537315 TI - The role of endogenous and exogenous nitric oxide on airway function. AB - Coordinated regulation of airway caliber is important for the maintenance of effective ventilation. Therefore, we sought to characterize the role of endogenously released, and exogenously administered, nitric oxide (NO) in mediating airway smooth muscle relaxation during early postnatal life. In both rat pup and piglet models, cholinergically mediated airway contractile responses were diminished by simultaneous release of endogenous NO and cGMP activation in both central airways and peripheral contractile elements. This ability of endogenously released NO to oppose airway constriction may be impaired in response to hyperoxic exposure or in animal models of cystic fibrosis. Additional studies in piglets have shown that exogenously administered NO causes a modest, but significant, reduction in lung resistance, analogous to the decrease in pulmonary vascular resistance induced by inhaled NO. Clinical trials are now underway in preterm infants at risk for chronic neonatal lung injury to determine whether inhaled NO has a beneficial effect on the development of bronchopulmonary dysplasia and whether exogenous NO modulates airway function in such infants. PMID- 12537317 TI - Genetic influences in respiratory distress syndrome: a twin study. AB - This is a twin study of the contribution of genetic influences on the pathogenesis of respiratory distress syndrome (RDS). Retrospectively, the files of twins born between 1976 and 1995 in the Academic Medical Centre (before Wilhelmina Gasthuis) of the University of Amsterdam were studied with a gestational age of 30 to 34 weeks and 1 or both with RDS. Data were collected on gestational age, birth weight, Apgar score, or diabetes in the mother as risk factors. All children were born vaginally. One hundred ninety-four pairs of twins were born with a gestational age of 30 to 34 weeks. Twenty-six pairs were excluded, because neither of the babies developed RDS. Of the 168 pairs, we were able to determine zygosity in 80 pairs, of which 18 were monozygotic and 62 were dizygotic twins. This is a normal ratio for the gestational age of 30 to 34 weeks. Risk factors as formulated above were not different between the monozygotic and dizygotic twins. RDS occurred more frequently in both twins when the twins were monozygotic (12 of 18, 67%) than when the twins were dizygotic (18 of 62, 29%). This difference was statistically significant, P < .05 by Chi square analysis. When only 1 twin developed RDS, it was the second born in 4 (67%) of the other 6 pairs of monozygotic twins and in 34 (77%) of the 44 dizygotic twins (77%). This twin study supports the notion of a genetic contribution to RDS. PMID- 12537316 TI - Nitrogen utilization and bone mineralization in very low birth weight infants fed partially hydrolyzed preterm formula. AB - Partialy hydrolyzed formulas have been proposed for term and preterm infants, but decreased nitrogen and calcium intestinal absorption rates, together with plasma amino acid imbalances have been reported in preterm infants. We evaluated a new formula with modified nitrogen and calcium sources (glycerophosphate). During their second week of life, 16 preterm infants were randomly assigned to 1 of 2 groups: 9 were fed the new partially hydrolyzed formula and 7 a conventional formula. A nutrient balance was performed at the end of the first month of life. Amino acid concentrations and whole-body mineralization were measured at the end of study period (theoretical term). Birth weight and gestational age (mean +/- SD) were similar in the 2 groups (28.9 +/- 7.0 wks and 1,183 +/- 242 g v 27.7 +/- 1.0 wks and 1,139 +/- 162 g). Median nitrogen absorption rate (85% v 89%; P = .03) was lower in infants fed the new formula than in those fed the conventional one. After correction for difference in nitrogen intake, there was no significant difference in N retained between the 2 groups (P = .11). Plasma amino acid concentrations were also similar in the 2 groups. At theoretical term, median bone mineral content was not significantly different between the 2 groups (50 g/kg v 55 g/kg; P = .17) and it was close to the reference values obtained in term neonates just after birth. As long as nitrogen content is 10% higher in protein hydrolyzed formula than in entire protein formula, appropriate nitrogen retention, plasma amino acid profile can be achieved with the new partially hydrolyzed formula. In both groups, bone mineralization at theoretical term was close to the mineralization observed term neonates just after birth. PMID- 12537318 TI - Surfactant proteins and genetic predisposition to respiratory distress syndrome. AB - Respiratory distress syndrome (RDS) is caused by surfactant deficiency at birth. The risk of RDS decreases from the gestational age of 24 weeks to full-term. Genetic and acquired factors additionally influence the risk of RDS. Surfactant deficiency in RDS is mainly caused by immaturity and a lack of differentiation of the alveolar epithelial cells involved in surfactant synthesis and secretion. A network of hormones and growth factors regulate perinatal development. Host related factors, including the levels of expression of surfactant proteins (SP), modulate the responsiveness of growth factors. SP-A has roles in surface activity and regulatory roles particularly in innate immunity; SP-B is essential for the processing of surfactant and for the surface activity; SP-C has roles in surfactant metabolism and function; the regulatory roles of SP-D mainly pertain to innate immunity. The genetic variation of SP-A and SP-B genes and the risk of RDS have been studied. Both SP-A and SP-B associate with susceptibility to RDS. The association between the SP-A allele and genotypes and the risk of RDS is dependent on the SP-B genotype and significantly influenced by the degree of prematurity, antenatal glucocorticoid therapy, multiple birth, and birth order. The alleles/genotypes of SP-A, SP-C, or SP-D also associate with several other inflammatory lung and airway diseases. Rare mutations in SP-B or SP-C cause serious, often fatal lung diseases. Genetic and post-genomic research is likely to eventually result in new diagnostic applications and specific therapies for the prevention of respiratory failure and inflammatory lung diseases. PMID- 12537319 TI - Is the BPD epidemic diminishing? AB - In a previous study of very low birth weight neonates, < or = 1500 g, admitted to the Vanderbilt University Neonatal Intensive Care Unit (NICU) from 1976-1990, improvements in survival were accompanied by a corresponding increase in the incidence of bronchopulmonary dysplasia (BPD). Since then, certain neonatal and perinatal interventions have been introduced and may influence neonatal outcomes. In this study, we have continued the analysis of the incidence of 3 outcomes: 1) Neonatal death (NEOD), 2) BPD, and 3) NEOD or BPD (NEOD/BPD) for an additional 7 years, 1991-1997. A retrospective study was performed of 3,837 patients with birth weight < or = 1500 g and admitted to the Vanderbilt NICU within 24 hours of birth from 1976 through 1997. The outcomes NEOD, BPD, or NEOD/BPD were modeled by using multiple logistic regression with the following risk factors included as covariates: birth weight, gestational age, Apgar scores at 1 and 5 minutes, gender, race, birth location, diagnosis of hyaline membrane disease, maternal age, maternal diabetes, delivery method, multiple births, duration of ruptured membranes, and biologically relevant interactions among these covariates. To assess time trends in the risk factors and outcomes, patients were divided into time periods (1 = 1976-80, 2 = 1981-85, 3 = 1986-90, 4 = 1991-95, and 5 = 1996 97). For each outcome, only covariates or interactions among covariates found to be significant were retained in the final model. Adjusted odds ratios and 95% confidence intervals were calculated to measure the risk associated with a given time period in comparison to the preceding period. There was a progressive decline in NEOD across all time periods. The previously described increase in BPD from period 1 through period 3 is followed by a decrease in periods 4 and 5. The risk of NEOD/BPD remained fairly constant from period 1 to period 3, but then showed a significant decrease over the two most recent periods. Prior to 1991, the cost of improved survival among very low birth weight infants in this large NICU was an increased incidence of BPD. Since 1991, the risk of BPD has been decreasing even though survival continues to improve. If these findings are also representative of other NICUs, they signify an important reduction in the impact of BPD as one of the costly sequelae of prematurity. PMID- 12537321 TI - Pathological gambling--a review of current clinical considerations. PMID- 12537320 TI - World report on violence and health. PMID- 12537322 TI - Drug expenditure in Ireland 1991-2001. PMID- 12537323 TI - Creating a safer Europe for our children. PMID- 12537324 TI - The six week postnatal check: women's choice of service provider. AB - This study describes mothers' choice between General Practitioner (GP) and hospital clinics for the six week postnatal check and elucidates influencing factors for their choice, using structured interviews with mothers within one week of discharge from hospital. Intended uptake of the six week check was high overall. For mother's own check, 203 (45%) planned to attend hospital clinics and 116 (26%) planned to attend the GP, while for the baby's check 115 (26%) planned to attend hospital clinics and 245 (55%) planned to attend the GP. The remainder planned to attend a consultant privately or had been referred to the specialised gynaecological/paediatric clinic. Factors influential for those choosing GPs were: convenience, familiarity and opportunity for discussion. Hospital instructions and the expected quality of examination were important for those choosing the hospital. Some possible misconceptions regarding provision of aspects of postnatal care were uncovered. This study highlights underuse of the GP service for the routine six week check and makes recommendations in relation to this. PMID- 12537325 TI - Evaluation of the Helicobacter pylori stool antigen (HpSA) test in routine clinical practice--is it patient-friendly? AB - A new immunoassay has been developed to detect the presence of Helicobacter pylori antigens in stool specimens. The aim of our study was to assess the accuracy and utility of the H. pylori stool antigen (HpSA) test in routine clinical practice. Dyspeptic patients undergoing endoscopy were studied. H. pylori status was defined before treatment by CLOtest and histology, and by 13C urea breath test (UBT) after eradication therapy. A standard universal container was provided for stool collection and the HpSA test was performed by an investigator blind to the results of the other diagnostic tests. Patients also provided a venous blood sample prior to endoscopy for H. pylori serology. Sixty patients (30 M : 30 F: mean age 47 yr) were enrolled in the study. The pretreatment sensitivity, specificity, positive and negative predictive value of the HpSA test were respectively, 93%, 94%, 96%, and 90%. Twenty five patients returned for post treatment 13CUBT, but only 14 (56%) provided a stool sample for analysis. The post treatment sensitivity, specificity, positive and negative predictive value of the HpSA test were respectively, 67%, 100%, 67%, and 92%. The HpSA test was negative in 19% of the patients found positive for anti-H. pylori antibodies on serology testing. All H. pylori antibody-negative patients had a negative HpSA test. Our results suggest that the HpSA test provides accurate pretreatment diagnosis of H. pylori infection but the reliability of the test after treatment is uncertain. A potential problem with the HpSA test appears to be patient reluctance about stool handling and this could prove a significant obstacle to patient compliance and the acceptability of the test in everyday clinical practice. PMID- 12537326 TI - Legionnaires' disease in Ireland--a cause for concern? AB - Legionnaires' disease is a multi-system illness that can have widespread clinical symptoms, though the principle manifestation is pneumonia. A review of all legionnaires' disease case notifications in Ireland in the 4 year period 1998 2001 was undertaken to assess the degree of concern health authorities here should have in relation to this disease. There were 17 cases of legionnaires' disease (all were legionella pneumophila serogroup 1) notified to the National Disease Surveillance Centre in the period 1998-2001. Nine cases were travel associated cases with the majority of cases occurring during the summer months. There were 2 deaths. It would appear that legionnaires' disease is rare in Ireland but when compared with other European countries Ireland is conspicuous by its low rate. This would suggest that a major degree of under diagnosis and under reporting of legionnaires' disease currently exists in Ireland. PMID- 12537327 TI - Late presentation of recurrent congenital tracheoesophageal fistula. PMID- 12537328 TI - Generic drug packaging leading to a critical incident. PMID- 12537329 TI - Carbon monoxide toxicity. PMID- 12537330 TI - Klippel-Trenaunay syndrome: an unusual cause of haematuria. PMID- 12537331 TI - Oaths & courses. PMID- 12537332 TI - Digit span changes from puberty to old age under different levels of education. AB - The goal of this study is to demonstrate the age-related changes in multimodality digit span under a research design in which level of education is controlled. Volunteer participants (n = 1183) were distributed over levels of age (13-98 years) and education (5-8, 9-11, and 12+ years). Digit span was measured through 11 scores of the Visual Aural Digit Span Test-Revised on aural or visual stimulation and oral or written response execution, thus allowing for the measurement of intra- and intersensory integration. The increase in digit span scores reversed to a decrease with early adulthood. The slope of the regression line was small but significant. A 4 x 3 x 2 multivariate analysis of variance showed a significant effect of age and education on a combined score comprising the 11 digit span scores. Differences of age and education were predicted by the auditory and visual input scores. The article discusses the cognitive correlates and the age-related changes in digit span from the biological standpoint. PMID- 12537333 TI - Memory functioning following traumatic brain injury in children with premorbid learning problems. AB - This study examines the memory functioning of 25 children who sustained a traumatic brain injury (TBI) and who had prior learning problems, 48 children with TBI who did not have prior learning problems, and 23 noninjured controls. The children with TBI and prior learning problems displayed significantly worse memory abilities than both the control participants and the children with TBI and no prior learning problems. They differed significantly from these 2 groups on measures of general memory, verbal memory, sound-symbol learning, and attention. The results suggest that children with premorbid learning problems who sustain TBI have less cognitive reserve and a lower threshold for the expression of cognitive impairments in areas that reflect preexisting learning and language problems, compared to children without premorbid learning problems. PMID- 12537334 TI - Electric brain responses obtained from newborn infants to changes in duration in complex harmonic tones. AB - Event-related brain potentials of 10 full-term newborns in response to duration changes in complex, harmonic tones known to elicit prominent mismatch negativity (MMN) response in adults were investigated. Here we report that duration changes elicited prominent MMN responses in a 11 newborns tested. In contrast, MMN has been reported in as little as 50% of the infants in some previous studies using other sound attributes. When the infant MMN latency was compared with that obtained from adults, and 4- as well as 8-year-old children, it was found that the MMN latency was a bit later in newborns than in older age groups. This result is consistent with previous findings. The MMN amplitude, however, was surprisingly large in infants compared to older children. PMID- 12537335 TI - Inhibition of prepotent responding and attentional flexibility in treated phenylketonuria. AB - Inhibition of prepotent responding and attentional flexibility were assessed in 58 early and continuously treated phenylketonuria (PKU) patients and 69 controls, age 7 to 14 years. A computerized task was used requiring participants to process consecutive stimuli according to various attentional sets. Analysis of error rate suggested poorer inhibition of prepotent responding in PKU patients compared with controls. No influence of concurrent plasma phenylalanine (phe) was shown, neither in the younger (age < 11 years) nor in the older participants (age > or = 11 years). Analysis of error rate provided strong evidence for poorer attentional flexibility in PKU patients compared with controls. The difference between attentional flexibility in controls and PKU patients could mainly be attributed to younger PKU patients, with concurrent phe levels higher than 360 micromol/L. Younger PKU patients with phe levels below 360 micromol/L performed at the same level as age-matched controls. Performance of PKU patients was strongly associated with phe levels in age periods during the first 10 years of life, which are characterized by a strong development of executive functioning (ages 2 7 and age 9). High phe levels during these age periods could delay development of inhibitory control and attentional flexibility. With regard to treatment, analyses with lifetime and concurrent phe levels support strict dietary control throughout the first decade of life, after which the phe-restricted diet can be relaxed. However, based on the evidence that development of specific executive functions continues until approximately age 12, it is recommended to maintain phe levels below 360 micromol/L throughout early adolescence. PMID- 12537336 TI - Neuropsychological evaluation of deficits in executive functioning for ADHD children with or without learning disabilities. AB - This study investigates multiple aspects of executive functioning in children with attention deficit/hyperactivity disorder (ADHD). These areas include attentional components, impulsiveness, planning, and problem solving. The rationale of the study is based on neurophysiological studies that suggest frontal lobe dysfunction in ADHD. As frontal lobe functioning is related to abilities in executive control, ADHD is hypothesised to be associated with deficits in various areas of executive functioning. The specific effect of comorbidity of learning disability (LD) was also investigated. Eighty-three children with ADHD and 29 age-matched controls (age 7-13) participated in the study. A battery of neuropsychological tests was utilized to evaluate specific deficits in speed of processing, selective attention, switching attention, sustained attention, attentional capacity, impulsiveness, planning and problem solving. Findings indicated that children with ADHD have slower verbal responses and sustained attention deficit. Deficits in selective attention and attentional capacity observed were largely related to the presence of LD. No specific deficit associated with ADHD or the comorbidity of LD was identified in switching attention, impulsiveness, planning, and problem solving. These results revealed that ADHD is not associated with a general deficit in executive functioning. Instead, ADHD is related to a specific deficit in regulation for attentional resources. The importance of isolating the deficit related to LDs for examining the specific deficit associated with ADHD is highlighted. Results also emphasised the importance of isolating the effect of lower level of abilities (e.g., speed of processing) and the utilization of specific definition for the examination of executive functions. PMID- 12537337 TI - Longevity of two-unit cantilevered resin-bonded fixed partial dentures. AB - PURPOSE: To examine the clinical performance of two-unit cantilevered resin bonded fixed partial dentures that were inserted at The Prince Philip Dental Hospital, Hong Kong by students and staff between 1992 and 1998. MATERIALS AND METHODS: A search of the Hospital computer records system revealed a list of 130 patients who had received two-unit cantilevered resin-bonded fixed partial dentures (RBFPD) placed more than 24 months previous to the review date. For each patient clinically examined, the following data were recorded: gender, age, operator, cementation date, endodontic treatment if performed, bone support, tooth mobility, the presence of shimstock contacts on abutment or pontic in intercuspal position, the presence of interproximal contacts adjacent to the prosthesis assessed by dental floss. Date of any debonds with subsequent treatment was recorded and the patient was asked qualitative questions about their prosthesis. RESULTS: 82 prostheses were placed in 69 patients and were found to have a mean service life of 36.7 +/- 15.4 months and a range of 4.3 months to 95.4 months. In total, four prostheses were reported to have debonded resulting in a clinical retention rate of 95.1%. No rotation, drifting or tipping was observed for any of the prostheses during the time of this study. Overall patients' satisfaction with RBFPDs was good with an average assessment rating of 8.2. PMID- 12537338 TI - Efficiency and cytotoxicity of resin-based desensitizing agents. AB - PURPOSE: To compare in vitro the efficacy of five resin-based desensitizing agents at reducing human dentin permeability and to compare their cytotoxicity. The tested hypothesis was that their different curing techniques cause variations in efficiency and cytotoxicity. MATERIALS AND METHODS: Dentin slices (0.5 +/- 0.05 mm thick) were prepared from human third molars (10 per group) and their hydraulic conductance was recorded before and after application of one of the desensitizing agents with a Flodec device. Six desensitizing agents were studied: one light curing agent (Seal and Protect); one self-curing agent (Pain Free); the resin-based agents without any polymerization initiator (Health-Dent, Gluma Desensitizer, Isodan); one oxalate-based agent served as a control (Protect). A MTT assay on L 929 fibroblasts was performed to measure the cytotoxicity of the six desensitizing agents applied onto additional dentin slices (10 per group). RESULTS: All the desensitizing agents resulted in a large decrease in dentin permeability. The best results were obtained with Gluma Desensitizer, Isodan, Pain Free and Protect. A statistically significant difference was found among the materials (P = 0.001). All the materials were non-cytotoxic. Cell viability ranged from 88% for Seal and Protect to 100% for Isodan. No difference was found among their cytotoxicity. PMID- 12537339 TI - Effect of phosphoric acid etching prior to self-etching primer application on adhesion of resin composite to enamel and dentin. AB - PURPOSE: To investigate the effect of phosphoric acid etching prior to the application of self-etching primer on the adhesion of resin composite to tooth substrates. MATERIALS AND METHODS: Bovine teeth were randomly divided in four groups of 20 samples each. Tooth surface conditions were as follows: Group 1: enamel ground with 600-grit SiC paper; Group 2: ground enamel was etched with 35% phosphoric acid gel for 15 seconds, water-rinsed and completely dried; Group 3: ground dentin; Group 4: dentin etched using the same method as Group 2. The samples in each group were divided in two subgroups of 10 each. UniFil Bond (UB) and Clearfil SE Bond (SE) were applied as adhesive systems with self-etching primers, and a layer of resin composite (AP-X) was placed and light-cured for 40 seconds. The tensile bond strengths (TBS) were measured and the resin-tooth interfaces were observed with scanning electron microscopy. RESULTS: The mean TBS values were 11.2 MPa (Group 1-UB), 14.3 MPa (Group 1-SE), 16.3 MPa (Group 2-UB), 20.5 MPa (Group 2-SE), 13.4 MPa (Group 3-UB), 16.7 MPa (Group 3-SE), 9.3 MPa (Group 4-UB) and 12.6 MPa (Group 4-SE). Two-way ANOVA and Scheffe's F test showed that the enamel etching significantly increased the TBS values but dentin etching significantly decreased the TBS values. PMID- 12537340 TI - Three-year clinical evaluation of One-Step in non-carious cervical lesions. AB - PURPOSE: To clinically evaluate the 3-year performance of a one-bottle dentin adhesive in the restoration of non-carious cervical lesions (NCCLs) with resin composite. MATERIALS AND METHODS: 105 NCCLs were restored with either AElite Flow, Bisco Glaze (both flowable resin-based composites) or Silux (a microfilled resin-based composite), using One-Step dentin adhesive. The cavities were cleaned with pumice and water slurry, the dentin lightly roughened with a fine diamond bur, and the restorations placed according to the manufacturer's instructions. Patients were recalled at 6 months, 1 year, 2 years and 3 years, and restoration integrity and marginal discoloration assessed. RESULTS: The overall retention rates were: 6 months, 100%; 1 year, 95%; 2 years, 87%; 3 years, 75%. Marginal discoloration was negligible, and there was no significant difference among the materials, although there was a trend for it to be greater around the Bisco Glaze restorations. PMID- 12537342 TI - Microleakage and damage to adjacent teeth when finishing Class II adhesive preparations using either a sonic device or bur. AB - PURPOSE: To detetmine the damage to adjacent tooth surfaces and the quality of the marginal seal of a resin-based composite restoration when a sonic preparation device (Sonicsys) was used to finish the preparation outline. MATERIALS AND METHODS: Three general practitioners prepared 60 Class II box-type cavities in extracted teeth placed in an artificial jaw in a phantom-head. In 30 preparations a bevel was made with a bur, in the other 30 restorations the margins were finished using the sonic preparation device. All preparations were restored with a total-etch technique and a posterior composite (SA primer-Photo Bond-Clearfil PhotoPosterior). Microleakage was tested and the adjacent teeth were inspected for iatrogenic damage. RESULTS: No statistically significant differences in microleakage were observed between the experimental and control groups. In the control group, 80% of the approximal surfaces of adjacent teeth were damaged while only 30% of the adjacent surfaces were damaged in the experimental group (P < 0.05, Chi-square test). PMID- 12537341 TI - Two-year clinical evaluation of Dyract in small Class I cavities. AB - PURPOSE: To evaluate the 2-year clinical performance of Dyract in small Class I cavities in non stress-bearing areas. MATERIALS AND METHODS: On 36 patients, 87 small cavity preparations were performed. The lesions were diagnosed macroscopically with a probe. They involved fissures, were shallow and had reached dentin but the lateral spread was limited and localized in dentin. Cavities were designed to be on non stress bearing areas. Cavities' average faciolingual widths were prepared to be 1/3 or less than the intercuspal width. At baseline, 1- and 2-year recalls, the restorations were evaluated according to the modified Ryge criteria by two calibrated, experienced examiners. RESULTS: None of the restorations was lost and retention rate was 100% at the end of 1 year. After 2 years, one restoration (1.1%) had to be replaced due to caries lesion adjacent to its margin and the rate of retention was 98.9%. At the 2-year recall, marginal discoloration (rated Bravo) was observed in 19.8% of the restorations but did not require the replacement of any restorations. Except the failed restoration, no other was clinically unacceptable in regard to color match, wear or loss of anatomic form, marginal discoloration, caries, marginal adaptation and surface texture. PMID- 12537343 TI - Effect of changing application times on adhesive systems bond strengths. AB - PURPOSE: To examine and compare the effect of increasing the priming time and the adhesive resin pre-curing time on shear bond strength of new dentin bonding systems (DBS). MATERIALS AND METHODS: Two multi-step/total etch DBS, Scotchbond Multi-Purpose Plus (SBMP+) and OptiBond FL (OB FL), and one one-bottle/total etch DBS, Single Bond (SB), were used. One hundred flat dentin specimens were prepared from extracted human molars. Approximately 1 mm of dental structure was ground to expose superficial dentin. The specimens were randomly assigned to the three DBS, and minimum of five specimens were used for each test condition. For each multi step DBS, 40 specimens were divided into four groups with priming times of 10, 20, 30, and 40 seconds. Each group was further subdivided into two subgroups with adhesive resin pre-curing times of 20 and 40 seconds. The SB specimens were divided into four subgroups with a waiting time of 10, 20, 30, and 40 seconds before drying and curing of its one-bottle adhesive. Shear bond strength tests were performed using a universal testing machine (Instron) at a crosshead speed of 0.5 mm/minute. Results were analyzed using ANOVA and Duncan's multiple range tests. RESULTS: For the multi-step DBS, increasing the priming time up to 30 or 40 seconds caused a significant increase in the mean shear bond strength values (P < 0.0001). Lengthening the adhesive resin pre-curing time did not cause any significant increase (P = 0.05) in the mean shear bond strength values except with 30-second and 40-second priming times. For SB, increasing the one-bottle adhesive waiting time before drying and curing to 30 seconds resulted in a statistically significant increase (P < 0.002) in the mean shear bond strength. PMID- 12537344 TI - Enhanced bond strengths of compomers using two dentin bonding systems. AB - PURPOSE: To measure bond strengths of three commercial compomers using recent dentin bonding systems, and to observe the micromorphology of the cross-sectional interface of the bonded specimens. MATERIALS AND METHODS: One hundred and ten bovine teeth stored frozen were used. The dentin surfaces were ground flat with #600-grit silicon carbide paper, and divided into three groups for bonding: original bonding system group (Xeno Bond/Xeno, Prime&Bond 2.1/Dyract AP, and F2000 Primer/Adhesive/F2000 compomer), Clearfil Liner Bond 2V group (LB 2V/Xeno, LB2V/Dyract AP, LB2V/F2000 compomer, LB2V/AP-X), and Single Bond group (SB/Xeno, SB/Dyract AP, F2000 compomer, SB/Z100). After the bonding procedures had been performed, the bonded assemblies were stored in water for 24 hours at 37 degrees C. Then, tensile bond strengths were measured using a universal testing machine at a crosshead speed of 2 mm/minute. All data were statistically analyzed using one-way ANOVA and Fisher's PLSD at 95% of confidence. Specimens for SEM observations were bonded in the same manner as for the tensile bond test, cut in half, and embedded in epoxy resin. These were then polished to high gloss, gold sputter-coated, and observed with the SEM. RESULTS: Compomers bonded to dentin with recent dentin bonding systems showed significantly higher bond strengths than those with the original bonding systems (P< 0.05). Tensile bond strength of Clearfil Liner Bond 2V was also significantly higher than that of Single Bond (P< 0.05). SEM observations showed good interaction between the compomers, adhesive systems, and dentin with any restorative material used. For Xeno Bond/Xeno, Prime & Bond 2.1/Dyract AP, and Primer/Adhesive/F2000 compomer, the thicknesses of the hybrid layer were approximately 0.8-1.1 microm, 0.3-0.5 microm, and 0.7-1.5 microm respectively. On the other hand, the hybrid layer produced by LB2V was approximately 0.5 to 1.0 microm, whereas for SB was approximately 3.1 to 4.1 microm thick. The thickness of the hybrid layers created in each bonded sample depended on the adhesive material used. PMID- 12537345 TI - Clinical evaluation of the role of glutardialdehyde in a one-bottle adhesive. AB - PURPOSE: To evaluate clinically whether the addition of glutardialdehyde to the one-bottle enamel-dentin adhesive Gluma Comfort Bond (GCB) has a desensitizing function, as insinuated by the brand name Gluma Comfort Bond+Desensitizer (GCB+D). MATERIALS AND METHODS: Two pain studies following identical protocols were conducted in Abu Dhabi (A) and Bologna (B). Each of the 60 and 59 patients selected, respectively, had two sensitive cervical tooth sites, characterized by a score three or higher on a five-step pain scale: 1 (no), 2 (slight), 3 (mild), 4 (severe), 5 (very severe) discomfort upon application of a 2-second cold air stimulus. The buccal sensitive sites treated in A were generally small, in B larger and in most cases extending into the proximal tooth area. The patients indicated their discomfort level according to the pain scale prior to and immediately after topical application of GCB and GCB+D, respectively, and then at recalls after 1 week, 1 (A) or 3 (B) months, and 6 months. The data was statistically analyzed by chi2-tests (P < 0.05). RESULTS: With the cumulative scores 1 and 2, the following percentage success rates were found for GCB/GCB+D. A: postoperatively 93/98, 1 week and 1 month 98/98, and at 6 months 92/100. In B, the respective decrease in sensitivity was uniformly 43/100 throughout the entire evaluation period, postoperatively through 6 months. The 33 teeth (57%) of GCB with scores 3 and 4 in the B-study received a rescue treatment with Gluma Desensitizer immediately after the 1 week recall. This regimen reduced the sensitivity instantaneously to scores 1 and 2 until the end of the evaluation after 6 months. In the A-study, GCB+D was significantly more effective than GCB after 1 week, 1 month, and 6 months, in the B-study at all recall sessions. It is hypothesized that a thin film of adhesive applied to a sensitive area may not polymerize and seal the dentin surface adequately due to oxygen inhibition. In particular, proximal sensitive tooth sites with poor access for adhesive application are suspected to remain inadequately sealed with a polymer film and thus sensitive. In this latter case however, glutardialdehyde as a component of the adhesive compound will result in coagulation of peripheral tubular liquid protein and thus desensitization, due to hampering of the liquid shift across the tubules upon excitation of a cold stimulus. In conclusion, glutardialdehyde played an active and effective role as a desensitizing agent, when added to the ethanol-water based one-bottle adhesive Gluma Comfort Bond. PMID- 12537346 TI - Inhibition of calculus formation by dentifrice formulations containing essential oils and zinc. AB - PURPOSE: To examine the effect of three essential oil-containing dentifrice formulations with different zinc salts and concentrations on dental calculus formation in rats. MATERIALS AND METHODS: A total of 150 rats were randomly distributed into five litter and sex-matched groups of 30. They were maintained on a calculogenic diet and topically treated twice daily with undiluted dentifrices. Each experimental dentifrice formulation contained a zinc salt, a fixed combination of essential oils (i.e. eucalyptol, menthol, methyl salicylate, and thymol) and a hydrated silica abrasive. Commercial zinc-free and zinc containing dentifrices were included as negative and positive controls, respectively. After 17 days calculus was scored post-mortem on molar teeth. RESULTS: All groups of rats treated with zinc-containing dentifrices had significantly (P < 0.05) lower mean calculus scores than the group treated with a zinc-free control dentifrice. Other variables (e.g. fomulation type, essential oil content, zinc salt and concentration) did not significantly affect calculus scores. Thus, all three essential oil/zinc-containing dentifrices and a commercial, zinc-containing, tartar-control dentifrice inhibited calculus formation in this rat model. PMID- 12537348 TI - Orthodontic and prosthodontic treatment in dental avulsion cases. AB - A clinical case of traumatic dental avulsion with endodontics, replantation and bracket fixation is presented. Subsequent follow-up led to extraction of the replanted teeth. The edentulous space was restored with a Maryland bridge. The factors which contribute to a successful tooth replantation as well as choosing the right time to begin orthodontic treatment in the case of tooth avulsion and replantation are described. PMID- 12537347 TI - Regeneration of soft and hard tissue periodontal defects. AB - Periodontitis is characterized by the formation of periodontal pockets and bone loss. Although the basic treatment emphasizes the control of bacterial plaque, the clinician is confronted with the need to correct soft and/or hard tissue defects that develop as a consequence of the disease. This article reviews the current status of regenerative approaches in treating soft and hard tissue defects (based mainly on findings from our own laboratory) and assessed the global applicability of these procedures. Many different techniques have been suggested to treat those defects with, in general, a high degree of success. From the present knowledge it can be concluded that periodontal soft and hard tissue regeneration is possible. Treatment of areas with localized gingival recession or insufficient keratinized gingiva can be achieved with soft tissue grafts or pedicle flaps, as well as with the use of dermal allografts. The treatment of hard tissue defects around teeth and implants can be approached using different types of bone grafts, guided tissue or bone regeneration, or a combination of these. The predictability of many of these therapies, however, still needs to be improved. Since most of these techniques are sensitive, specific, and expensive, their present universal application is limited. PMID- 12537350 TI - Malate dehydrogenases--structure and function. AB - Malate dehydrogenases (MDH, L-malate:NAD oxidoreductase, EC 1.1.1.37), catalyze the NAD/NADH-dependent interconversion of the substrates malate and oxaloacetate. This reaction plays a key part in the malate/aspartate shuttle across the mitochondrial membrane, and in the tricarboxylic acid cycle within the mitochondrial matrix. They are homodimeric molecules in most organisms, including all eukaryots and the most bacterial species. The enzymes share a common catalytic mechanism and their kinetic properties are similar, which demonstrates a high degree of structural similarity. The three-dimensional structures and elements essential for catalysis are conserved between mitochondrial and cytoplasmic forms of MDH in eukaryotic cells even though these isoenzymes are only marginally related at the level of primary structure. PMID- 12537349 TI - Mitogen-activated protein kinases and their role in regulation of cellular processes. AB - Mitogen-activated protein kinases (MAPKs) are evolutionary conserved enzymes connecting cell-surface receptors to critical regulatory targets within cells. The three major MAPK cascades are known, the extracellular signal-regulated protein kinase (ERK) cascade, c-Jun amino-terminal protein kinase/stress activated protein kinase (JNK/SAPK) cascade and p38-MAPK cascade. This paper is focused on characterization of these MAPK cascades in terms of their distribution and biological role in some pathological processes (apoptosis, hypertrophy) with a special orientation on the role of MAPKs in cardiovascular system during ischemia/reperfusion. PMID- 12537351 TI - Glycoproteins of chicken liver nuclei cross-linked to DNA by cis diamminedichloroplatinum. AB - Glycoproteins which participate in DNA-protein cross-links induced by action of cis-diamminedichloroplatinum (cis-DDP) in intact nuclei of chicken liver were investigated. Digoxigenin-labelled lectins with different sugar specificity were used for detection and characterization of these glycoproteins. Our results showed the presence of glycoproteins bearing high mannose as well as complex type oligosaccharides in chicken liver nuclei. In most cases of complex oligosaccharides, sialic acid residues bound in alpha(2-6) but not in alpha(2-3) linkage were present. PMID- 12537352 TI - Effects of mechanical interaction between two rabbit cardiac muscles connected in parallel. AB - The hypothesis that myocardium mechanical inhomogeneity produces a substantial effect on mechanical function was tested. Muscle inhomogeneity was studied in isolated papillary muscles or trabeculae excised from rabbit right ventricle and connected in a parallel duplex. Each muscle was placed in a separate perfusion bath. One end of each muscle was fastened to an individual force transducer and the other to the common lever of a servomotor. This arrangement allowed both muscles, being excited independently, to pull jointly a load applied to the lever. Separate electrodes for each perfusion bath allowed to stimulate muscles with a time delay. Tension developed in the individual muscles and their interaction were studied. Developed tension was critically dependent on the timing and sequence of excitation. Using mathematical modeling, patterns of tension distribution experimentally observed in parallel duplexes were simulated. These results suggest that changes both in Ca(2+) transients and in the time course of Ca(2+)-troponin complexion due to the duplexed muscles interaction offset the effect of mechanical inhomogeneity. PMID- 12537354 TI - Three electrophysiological phenotypes of cultured human umbilical vein endothelial cells. AB - The conventional whole cell patch-clamp technique was used to measure the resting membrane conductance and membrane currents of nonstimulated cultured human umbilical vein endothelial cells (HUVECs) in different ionic conditions. Three electrophysiological phenotypes of cultured HUVECs (n = 122) were determined: first, 20% of cells as type I mainly displaying the inwardly rectifying potassium current (IKi); second, 38% of cells as type II in which IKi was super-posed on a TEA-sensitive, delayed rectifying current; third, 27% of cells as type III predominantly displaying the outwardly rectifying current which was sensitive to TEA and slightly inhibited by a chloride channel blocker niflumic acid (N.A.). In cells of type I, the mean zero-current potential (V0) was dependent on extracellular K+ ([K+]o) but not on Cl-, indicating major permeability to K+. Whereas V0 of type II was also affected by extracellular Cl- ([Cl-]o), indicating the contribution of an outward Cl- current in setting V0. The cells of type III were not sensitive to decrease of [Cl-]o and the outward current was activated in a relative stable voltage range. This varying phenotypic expression and multipotential behavior of HUVECs suggests that the electrical features of HUVEC may be primarily determined by embryonic origin and local effect of the microenvironment. This research provided the detailed electrophysiological knowledge of the endothelial cells. PMID- 12537353 TI - Sodium and ATP affinities of the cardiac Na(+),K(+)-ATPase in spontaneously hypertensive rats. AB - The Na(+),K(+)-ATPase is postulated to be involved in systemic vascular hypertension through its effects on smooth muscle reactivity and cardiac contractility. Investigating the kinetic properties of the above enzyme we tried to assess the molecular basis of alterations in transmembrane Na(+)-efflux from cardiac cells in spontaneously hypertensive rats (SHR). In the investigated group of SHR the systolic blood pressure and the heart weight were increased by 48% and by 60%, respectively. Upon activating the cardiac Na(+),K(+)-ATPase with substrate, its activity was lower in SHR in the whole concentration range of ATP. Evaluation of kinetic parameters revealed a decrease of the maximum velocity (Vmax) by 28% which was accompanied with lowered affinity of the ATP-binding site as indicated by the increased value of Michaelis-Menten constant (Km) by 354% in SHR. During activation with Na(+), we observed an inhibition of the enzyme in hearts from SHR at all tested Na(+) concentrations. The value of Vmax decreased by 37%, and the concentration of Na(+) that gives half maximal reaction velocity (KNa) increased by 98%. This impairment in the affinity of the Na(+)-binding site together with decreased affinity to ATP in the molecule of the Na(+),K(+)-ATPase are probably responsible for the deteriorated efflux of the excessive Na(+) from the intracellular space in hearts of SHR. PMID- 12537355 TI - Mechanisms of iron-induced oxidative modifications of creatine kinase in rat brain in vitro. possible involvement of HNE. AB - The model of oxidative stress induced by Fe/ascorbate in rat brain in vitro was used to compare the antioxidant capacity of known antioxidants. Creatine kinase (CK) was selected as a marker of protein injury in such studies. Of the antioxidant enzymes (catalase, superoxide dismutase), oxygen radical scavengers (mannitol, glutathione), and the chelator (EDTA) tested in this work and this system, only catalase and glutathione prevented the injury induced by oxidative stress, indicating that H2O2 and the glutathione peroxidase reaction were involved in the preventive effect. Additionally, the preventive effect of glutathione may be caused also by the fact that glutathione easily reacts with 4 hydroxynonenal (HNE), generated in rat brain homogenate, thus protecting CK from inactivation by this aldehyde. To find out whether and if at which concentrations CK may be oxidatively modified by HNE, pure CK was incubated in the presence of 10 and 64 micromol/l HNE for 30 min at 37 degrees C. The activity of CK incubated with HNE decreased significantly. Simultaneously, the protein carbonyls, determined by electrophoresis and immunoblotting increased at 10 micromol/l HNE or disappeared probably due to crosslinking of CK at 64 micromol/l HNE. The concentration of HNE in rat brain homogenates after oxidative stress was determined by HPLC and was in the range of 10-16 nmol/mg prot., corresponding to a concentration of 10-16 micromol/l HNE. This indicates that CK of rat brain homogenates oxidized by Fe/ascorbate may be impaired not only directly by oxygen radicals but also secondarily by HNE. PMID- 12537357 TI - Chemometric contributions to the evolution of metabonomics: mathematical solutions to characterising and interpreting complex biological NMR spectra. AB - The pharmaceutical industry has embraced emerging technologies such as genomics, proteomics and metabonomics over the past decade with a view to minimizing attrition and expanding drug development pipelines. Metabonomic technology, based on the multivariate analysis of complex biological profiles generated from spectroscopic instruments, has enabled the construction of successful expert systems for toxicity screening and disease diagnosis. Here we describe the evolution of chemometric and bioinformatic methodologies to accommodate the multi and megavariate data generated by high resolution NMR spectroscopy of biofluids, tissues and cell cultures and explore their potential role in mining, modeling and predicting metabolic data. PMID- 12537358 TI - A simple PDMS-based electro-fluidic interface for microchip electrophoretic separations. AB - High voltage electrodes for electrophoresis have been integrated into a polymer layer that can be reversibly bound to glass microchips for electrophoretic separations. By using the liquid precursor to the polymer polydimethylsiloxane (PDMS), platinum electrodes and reservoirs can be positioned prior to solidification, providing a simple and flexible method for electrode interface construction. Field strengths up to 875 V cm(-1) over an 8 cm separation channel can be applied to the system without any loss in performance of the interface. The interface can function as an electro-fluidic interface between the high voltage power supply and the separation channel and, when reversibly sealed to an etched glass plate, functions as a cover plate establishing a hybrid PDMS-glass microchip in which the electrodes are directly integrated onto the device. The versatility of this approach is not only demonstrated by separating DNA fragments in a novel buffer sieving matrix, but also with the molecular diagnostic analysis of a variety of DNA samples for Duschenne Muscular Dystrophy and cytomegalovirus (CMV) infection, using both microchip interface configurations. PMID- 12537359 TI - Performance of a simple UV LED light source in the capillary electrophoresis of inorganic anions with indirect detection using a chromate background electrolyte. AB - Light emitting diodes (LEDs) are known to be excellent light sources for detectors in liquid chromatography and capillary electromigration separation techniques, but to date only LEDs emitting in the visible range have been used. In this work, a UV LED was investigated as a simple alternative light source to standard mercury or deuterium lamps for use in indirect photometric detection of inorganic anions using capillary electrophoresis with a chromate background electrolyte (BGE). The UV LED used had an emission maximum at 379.5 nm, a wavelength at which chromate absorbs strongly and exhibits a 47% higher molar absorptivity than at 254 nm when using a standard mercury light source. The noise, sensitivity and linearity of the LED detector were evaluated and all exhibited superior performance to the mercury light source (up to 70% decrease in noise, up to 26.2% increase in sensitivity, and over 100% increase in linear range). Using the LED detector with a simple chromate-diethanolamine background electrolyte, limits of detection for the common inorganic anions, Cl-, NO3-, SO4(2-), F- and PO4(3-) ranged from 3 to 14 microg L(-1), using electrostatic injection at -5 kV for 5 s. PMID- 12537356 TI - Effects of aminoguanidine Schiff's base on biomarkers of the oxidative stress, 4 hydroxy-2-nonenal and conjugated dienes, in the model diabetes mellitus. AB - Diabetes mellitus evoked by streptozotocine in rats is associated with the oxidative stress. We examined the effect of Schiff's base 2,5 dihydroxybenzaldehyde with a well-known antidiabetic drug aminoguanidine, 2,5 dihydroxybenzilideneaminoguanidine (BAG) on the production of markers of oxidative stress such as 4-hydroxy-2-nonenal (4HNE) and conjugated dienes in diabetic rats. BAG administration did not affect glucose level in diabetic rats but significantly decreased the production of 4HNE and conjugated dienes. On the other hand, BAG caused the elevation of conjugated dienes and an insignificant increase of 4HNE levels in the control animals. PMID- 12537360 TI - Homogeneous chemical derivatisation of carbon particles: a novel method for funtionalising carbon surfaces. AB - The homogeneous reduction of diazonium salts by hypophosphorous acid, provides an easy methodology for the derivatisation of carbon powder. The resulting derivatised carbon was characterised electrochemically showing the behaviour expected of a surface bound species. This procedure provides an easy, inexpensive methodology for functionalising carbon which might find applications in ion exchange resins or combinatorial chemistry. PMID- 12537361 TI - Selective amperometric detection of dopamine using OPPy-modified diamond microsensor system. AB - Highly boron-doped diamond microfiber electrodes (BDDMF) were fabricated and characterized by the use of Scanning Electron Microscopy (SEM), Raman spectroscopy, and cyclic voltammetry. Amperometric detection of dopamine (DA), a neurotransmitter was achieved at pH 7.0, using BDDMF electrodes. The interferences from ascorbic acid (AA) and DOPAC were efficiently eliminated by using overoxidized polypyrrole-modified BDDMF electrodes, which also increased the sensitivity for the detection of dopamine. The limit of detection (S/N = 3) for dopamine was 0.1 nM, which is one order lower than that observed for carbon microfiber electrodes (CMFE), and the linear dynamic range was obtained from 0.5 nM to 100 microM (r2 = 0.997). The amperometric response for 0.5 nM dopamine has shown high stability with an RSD of 5.4% (n = 5). Highly reproducible results were obtained with an RSD of 6.2% for 10 measurements of 1 nM DA taken during 10 h and also remained the same, during measurements for 7 days, with no variation in efficiency for rejection of AA and DOPAC. PMID- 12537362 TI - Stepwise surface regeneration of electrochemical immunosensors working on biocatalyzed precipitation. AB - A new strategy of stepwise surface regeneration for electrochemical immunosensors, working on a biocatalyzed precipitation reaction, has been developed. The strategy is based on the combination of deposited product thin film dissolution and bound-protein displacement reactions from the modified sensor surfaces. As a model system, surfaces functionalized with biotin groups and their affinity recognition/ displacement reactions with antibiotin antibody molecules were chosen and investigated for affinity-sensing and stepwise regeneration reactions. PMID- 12537363 TI - Case-based reasoning (CBR) for multicomponent analysis using sensor arrays: application to water quality evaluation. AB - This paper reports by the first time the use of a simple, convenient and improved method of artificial intelligence (case-based reasoning, CBR) for multicomponent analysis, involving the data treatment afforded by electrochemical sensor arrays. The suitability of this method has been shown in aqueous solutions containing mixtures of ions of different nature and concentration, being applied in this case to a rapid recognition and evaluation of mineral water samples chosen as an illustrative example. The convenience of such a procedure opens the way to dispose of a versatile tool applicable for obtaining analytical information about ions present in a wide range of other multicomponent systems, and also to ascribe properties inherent to the examined samples. PMID- 12537364 TI - Real-time monitoring of lactate extrusion and glucose consumption of cultured cells using a lab-on-valve system. AB - Microsequential injection (microST) provides microfluidic operations that are ideally suited for cellular function studies and as a means of validating targets for drug discovery. MicroSI carried out within the lab-on-valve (LOV) manifold, is an ideal platform for spectroscopic studies on living cells that are grown on microcarrier beads and kept thermostated while their metabolism is probed in real time. In this paper a microbioreactor is integrated into the LOV manifold allowing measurement of cellular lactate extrusion and glucose consumption rates of a cell culture that is automatically renewed prior to each measurement. Glucose consumption and lactate extrusion are monitored using NAD-linked enzymatic assays. The microSI-LOV setup has demonstrated a linear analysis range of 0.05-1.00 mM for lactate and 0.1-5.6 mM for glucose. These assays were conducted in a serial fashion requiring 3 microL of cellular perfusate and 10 s for glucose determination and 30 s for the lactate assay. Overall waste generated per lactate/glucose assay is < 200 microL. This work was performed using two different transfected hepatocyte cell lines, which adhere to Cytopore microcarrier beads. This novel approach to metabolic screening allows for the rapid evaluation of the effects of dosing cells with chemical agents. PMID- 12537365 TI - The characterization of biomolecular secondary structures by surface plasmon resonance. AB - Recently there has been considerable interest in using surface plasmon resonance (SPR) for the measurement of conformational changes of immobilized biomolecules that are induced by an exogenous analyte. While a number of studies have shown the analytical utility of such measurements, there has been no report which characterizes the specific secondary structure that actuates the change in SPR signal. The use of SPR to indicate the type of secondary structure present in two immobilized polypeptides, poly-L-lysine (PL) and poly-L-glutamic acid (PGA), and a globular protein, concanavalin A (Con A) is described in this report. The PL, PGA and Con A were modified with N-succinimidyl 3-(2-pyridyldithiol) propionate (SPDP) to introduce disulfide groups to facilitate the attachment onto gold coated surfaces via self-assembly. Ethanol and 2,2,2-trifluoroethanol (TFE) were used to induce changes in the secondary structure of the immobilized polypeptides and the protein respectively. Using both circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopies, it has been demonstrated that it is possible to correlate the signal changes observed in SPR to the secondary conformation of the biomolecule. Both CD and FTIR showed that a decrease in SPR signal corresponded to a high content of beta, turn or unordered structures while an increase corresponded to a high alpha-helical content. The sensitivity of the SPR technique is comparable to that obtained in solution with CD and FTIR spectroscopies. These results are the first demonstration that SPR can be used to characterize secondary structures. There is potential, therefore, for SPR to be used as a technique to study secondary conformational changes of immobilized polypeptides and proteins. PMID- 12537366 TI - Acoustic coupling of transverse waves as a mechanism for the label-free detection of protein-small molecule interactions. AB - An on-line acoustic transverse wave device has been used to study the binding interactions of human serum albumin with the small molecule drug, warfarin. Four linking systems for the covalent attachment of the protein to the surface of the gold electrode of the sensor were employed, namely thioctic acid, cysteamine, an N-hydroxysuccinimide ester and 11-mercaptoundecanoic acid. All the attachment protocols involve the ability of thiols to form gold-sulfur bonds at the metal surface. The functional group present at the distal end of each thiol was chemically activated in order to facilitate covalent attachment of the protein. On-line sensor measurements of acoustic parameters show that the binding of warfarin to the protein can be detected, and depending on the linking monolayer used three of four possible combinations of changes in series resonance frequency and motional resistance are observed. Calculations of possible mass and thickness viscoelastic effects demonstrate that these conventional notions are invalid in terms of an explanation of the acoustic signals observed for the warfarin-protein interaction. The responses are ascribed to acoustic coupling phenomena. PMID- 12537367 TI - Headspace solid-phase microextraction--comprehensive two-dimensional gas chromatography of wound induced plant volatile organic compound emissions. AB - Plant emissions of volatile organic compounds from mechanically wounded Agrostis stolonifera, Pennisetum clandestinum, Eucalyptus leucoxylon and Trifolium repens have been sampled by headspace-solid phase microextraction (HS-SPME) and analysed by using comprehensive two-dimensional gas chromatography (GCxGC) for measurement of the plant emissions. GCxGC produces a fingerprint of the volatile organic compounds in a 2D separation space that may be approximately interpreted as a boiling point-polarity space, and may then be presented as a two-dimensional contour plot. This allows identification of sample-dependent variations in component distributions in the 2D plot, which will contain information about plant differences and should therefore facilitate recognition of different plant materials and displays the gross differences in volatiles between each plant species. PMID- 12537368 TI - Selectivity differences between sol-gel coated and immobilized liquid film open tubular columns for gas chromatography. AB - The solvation parameter model is used to determine the system constants for two sol-gel coated open-tubular columns at five equally spaced temperatures in the range 60-140 degrees C. Differences in the system constants as a function of temperature are used to determine the affect of sol-gel structure on the selectivity of SolGel-l and SolGel-Wax columns compared with conventionally coated and immobilized poly(dimethylsiloxane) and poly(ethylene glycol) stationary phases. The sol-gel columns should be suitable for similar separations to those presently performed on conventional immobilized liquid film columns of the same type but selectivity differences for polar compounds, which depend on temperature, should be anticipated. PMID- 12537369 TI - Anion exchanger as a reaction/separation medium-absorptiometric determination of trace amounts of boron in waters by on-line complexation with chromotropic acid presorbed on the anion-exchange column. AB - A novel method of on-line absorptiometric determination for trace amounts of boron was developed based on the complexation with chromotropic acid presorbed on an anion-exchange column. On-line reaction and separation were achieved by controlling pH conditions in solutions to accelerate the 1:2 complex formation in the concentration process at pH 3 and to stabilize the complex in the separation process at pH 8. About 75% of the boron introduced into the stream was kinetically collected as the 1:2 complex on the column under the experimental conditions. The 1:2 complex was satisfactorily separated from excess reagent and matrix components by changing the concentration of NaClO4 in the eluent and its peak height on the chromatogram monitored at 350 nm was used for calibration. The sensitivity could be enhanced by increasing the sample amount introduced and the detection limits (3sigma) were 162 ng dm(-3) and 45 ng dm(-3) of boron, when 5.0 cm3 and 13.4 cm3 of the samples were used, respectively. The method has been successfully applied to the determination of boron in samples of river water, tap water and ion-exchanged water. PMID- 12537370 TI - Optimized separation and determination of methyl sulfone metabolites of polychlorinated biphenyls (PCBs) and p,p'-DDE in biota samples. AB - An optimised method is described for the determination of 27 methyl sulfone polychlorobiphenyls (PCBs) and DDE in biota samples. Initially, the samples were extracted by hot Soxhlet and the methyl sulfones were separated by liquid/liquid extraction with concentrated sulfuric acid and back-extracted with hexane. The parameters of the back-extraction were studied and it was found that for a quantitative extraction of the methyl sulfones from the concentrated acid layer, a 50% dilution with cold water should be done. The hexane layer containing the methyl sulfones was further cleaned-up on basic silica (33% KOH) and Florisil. After concentration, the extract was analysed by gas chromatography-mass spectrometry (GC-MS) with electron capture negative ionisation (ECNI) in selected ion monitoring mode (SIM). It was shown that, for methyl sulfones, the ion formation was dependent on the chlorine substitution, position of the MeSO2-group and the ion source temperature. If the ion source temperature was higher than 200 degrees C, [M-CH3]- was the predominant ion for most methyl sulfones. Therefore, for increased sensitivity, quantitation of most congeners was done using [M-CH3]- ions instead of the molecular ion as used in previously reported methods. The method was validated for the determination of 26 tri- to hepta- 3- and 4 substituted MeSO2-PCBs and 3-MeSO2-DDE in animal and human tissues. Good sensitivity and selectivity of the method were obtained. Limits of detection (LODs) ranged from 0.06 to 0.10 ng g(-1) lipid weight. Average recoveries of individual congeners from vegetable oil spiked with individual standards (3.33 ng g(-1)) ranged from 73 to 112% with a mean value of 89%. The coefficients of variation ranged from 5.2 to 12.2%, which is within the acceptable range for environmental analyses. PMID- 12537371 TI - Development of an LC/MS method for the trace analysis of triacetone triperoxide (TATP). AB - The detection and quantification of triacetone triperoxide (TATP) using LC/MS is investigated. GC/MS analysis of TATP is hindered by stationary phase activation in very short periods of time. Due to the lower temperatures used in LC. this problem is not encountered. This study presents a method that is suitable for the detection of TATP at levels as low as 100 pg microl(-1) (10 ng per 100 microl). Initial findings are also reported for the investigation of a secondary chromatographic peak, which is thought to be caused by separation of two conformers. This study concludes that LC/MS is a suitable technique for the analysis of trace levels of TATP. PMID- 12537372 TI - Competitive immunoassay for vancomycin using capillary electrophoresis with laser induced fluorescence detection. AB - A competitive immunoassay using capillary electrophoresis with laser-induced fluorescence was developed for vancomycin. Capillary electrophoresis using a Tris glycine running buffer provided adequate separation of the antibody-bound from the unbound fluorescent probe (tracer) in less than 4 min. Laser-induced fluorescence polarization (LIFP) provided high sensitivity detection and simultaneous monitoring of fluorescence intensity and polarization. A fluorescence polarization value of 0.30 confirmed the formation of the antibody tracer complex. Calibration curves showed a working linear range of 2-3 orders of magnitude with a minimum detectable concentration of 0.98 ng mL(-1) (or 1.1 fg vancomycin). Clinical samples obtained from patients undergoing vancomycin treatment were analyzed for vancomycin and the results correlated well with a standard immunoassay based on latex particle detection that was routinely used by a hospital laboratory. Only 1/10 of the reagents were needed as compared with the standard immunoassay. PMID- 12537373 TI - Development of a sequential enzymolysis approach for the evaluation of the bioaccessibility of Cd and Pb from cocoa. AB - An in vitro model simulating enzymatic activity in the gastrointestinal tract was developed for the assessment of the potential bioaccessibility of Cd and Pb in cocoa powder and liquor. The model was based on the sequential extraction with simulated gastric and intestinal juices; the residue after the latter extraction was further investigated by using, in parallel, solutions of phytase and cellulase. The solubility of Cd and Pb in the corresponding enzymatic extracts was measured by ICP MS. The bioaccessibility of Cd in cocoa varied from 10 to 50% in gastrointestinal conditions. An additional 20 or 30% of Cd could be recovered by phytase and cellulase, respectively. The bioaccessibility of Pb in gastrointestinal conditions did not exceed 5-10%. Only a few percent more of this metal could be recovered by extraction with phytase and cellulase. PMID- 12537374 TI - Evaluation of the responses of a semiconductor gas sensor to gaseous mixtures under the application of temperature modulation. AB - A novel gas-sensing system based on a dynamic nonlinear response is reported to evaluate the effect of gaseous mixtures on the sensor response. A sinusoidal temperature perturbation was applied to a semiconductor gas sensor and the resulting conductance of the sensor was analyzed by fast Fourier transformation (FFT). The sensor response, which changed characteristically depending on the composition of the gaseous mixture, could be classified into three types (enhanced, suppressed, and preferential responses) by the mixture. To monitor the progress of the reaction of gases, the sample gas was analyzed by gas chromatography. The coexistent effect on the response to gaseous mixtures was theoretically simulated by considering the kinetics of gas molecules on the semiconductor surface. PMID- 12537375 TI - Renewable-surface sol-gel derived carbon ceramic electrode fabricated by [Ru(bpy)(tpy)Cl]PF6 and its application as an amperometric sensor for sulfide and sulfur oxoanions. AB - A highly sensitive and fast responding sensor for the determination of thiosulfate, sulfite, sulfide and dithionite is described. It consists of a chemically modified carbon ceramic composite electrode (CCE) containing [Ru(bpy)(tpy)Cl]PF6 complex that was constructed by the sol-gel technique. A reversible redox couple of Ru(II)/Ru(III) was observed as a solute in acetonitrile solution and as a component of carbon based conducting composite electrode. Electrochemical behavior and stability of modified CCE were investigated by cyclic voltametry, the apparent electron transfer rate constant (kappa(S)) and transfer coefficient (a) were determined by cyclic voltametry which were about 28 s(-1) and 0.43 respectively. Electrocatalytic oxidation of S(2-), SO3(2-), S2O4(2-) and S2O3(2-) were effective at the modified electrode at significantly reduced overpotentials and in the pH range 1-11. Optimum pH values for amperometric detection of thiosulfate, dithionite, sulfide and sulfite are 7, 9, 2 and 2. Under the optimized conditions the calibration curves are linear in the concentration ranges 1-500, 3-80, 2-90 and 1-100 microM for S2O3(2-), SO3(2 ), S2- and S2O4(2-) determination. The detection limit (signal to noise is 3) and sensitivity are 0.5 and 12, 2.8 and 6, 1.6 and 8, and 0.65 microM and 80 nA microM(-1) for thiosulfate, sulfite, sulfide and dithionite detection. The modified carbon ceramic electrode doped with Ru-complex shows good reproducibility, a short response time (t < 2 s), remarkable long term stability (> 6 month) and especially good surface renewability by simple mechanical polishing (RSD for eight successive polishing is 2%). The advantages of this sulfur compound amperometric detector based on ruthenium doped CCE are high sensitivity, inherent stability at a broader pH range, excellent catalytic activity, less expense and simplicity of preparation in comparison with recently published papers. This sensor can be used as a chromatographic detector for analysis of sulfur derivatives. PMID- 12537376 TI - Speciation analysis of aluminium(III) in natural waters and biological fluids by complexing with various catechols followed by differential pulse voltammetry detection. AB - The biological effects of aluminium have received much attention in recent years. Speciation of Al is of basic relevance as it concerns its reactivity and bioavailability. A differential pulse voltammetry (DPV) procedure is proposed for speciation analysis of Al(III) in natural waters and biological fluids using six catechols (L-dopa, dopamine, epinephrine, norepinephrine, caffeic acid and o benzenediol) as electroactive ligands. The decrease of the DPV anodic peak current for each catechol ligand is linear with the increase of Al concentration. This speciation analysis idea is based on the measurement of the complexation capacity, namely, different affinities of Al(III) for catechols and organic ligands under two pH conditions. The labile monomeric Al fraction (mainly inorganic aluminium) is determined at pH 4.6, while the total monomeric Al fraction is determined at pH 8.5. The principle for Al(III) speciation analysis by an electrochemical method is discussed. This sensitive and simple fractionation method is successfully applied to the speciation analysis of Al in natural waters and the results agree well with those of Driscoll's method. The speciation analysis of Al in biological fluids is also explored and the results are compared with those obtained by ultrafiltration and dialysis. Compared with other speciation protocols the electrochemical method possesses some remarkable advantages: rapidity, high sensitivity, cheap instrumentation and a simple operation procedure. PMID- 12537377 TI - A comparison of the Kjeldahl and Dumas methods for the determination of protein in foods, using data from a proficiency testing scheme. AB - Both the Kjeldahl and the Dumas methods for the determination of protein in foodstuffs are currently in use, but the empirical nitrogen factors used to convert the determined nitrogen content to protein content are based on the Kjeldahl method alone. Non-equivalence between the two methods could therefore result in some laboratories reporting an incorrect protein content. We report here a study using data accumulated over several years in the results of a proficiency testing scheme. On average the Dumas method provided results that were relatively higher by about 1.4% than the Kjeldahl method, but the difference between the methods depended on the type of foodstuff. The methodology of looking for bias between analytical methods is critically discussed. PMID- 12537378 TI - Precision estimates produced by specially-designed ruggedness tests compared with those derived from collaborative trials, in relation to estimation of measurement uncertainty. AB - We have compared the reproducibility precisions provided by collaborative trials with precisions obtained from ruggedness tests specially designed to try to simulate between-laboratory variation. We found that the ruggedness tests underestimated the reproducibility variability consistently for those analytical methods requiring empirical calibration. The ruggedness tests provided on average a precision merely comparable with repeatability precision. This finding has implications for the estimation of uncertainty. PMID- 12537379 TI - From experimental design to uncertainty estimation for the European Pharmacopoeia HPLC analysis of human insulin. AB - In this paper the process from experimental design (e.g. ruggedness test) to uncertainty estimation is described. The uncertainty estimate was calculated for the peak area of insulin plus A21 desamido insulin resulting from an HPLC analysis of a sample of an injectable human insulin preparation, Actrapid HM 100 IU ml(-1) (Novo Nordisk A/S). The analytical method used was the European Pharmacopoeia assay. (4) An expanded uncertainty (1) of 1.8% (of the area from the HPLC analysis) at an approximately 95% confidence level was found and confirmed by a validation study. This uncertainty refers to the peak area of the analyte from a single injection of the sample. The input parameters to the uncertainty estimate were found from a factorial experimental design (e.g. ruggedness test) consisting of 9 factors applied to the HPLC analysis. The input parameters were chosen to cover the probable contributors to the variability of the measured area of the HPLC analysis, including the sample preparation, but excluding uncertainty deriving from the reference material(s). PMID- 12537380 TI - The transfer between instruments of a reflectance near-infrared assay for paracetamol in intact tablets. AB - This study compares several correction methods to facilitate the transfer of a validated near-infrared (NIR) assay for paracetamol in intact tablets between two reflectance NIR instruments of the same type. Transfer was defined as the ability to accurately predict the true assay value of a sample measured on a NIR system using an assay developed on a different system, and was assessed using a comprehensive set of statistical tests. Direct electronic transfer of the calibration models, representing the NIR assay, was not possible as a result of a definite residual spectrum between instruments. The use of a correction method based on the standardisation of the material used to record the reference spectrum also proved ineffective. Two methods investigated did succeed, the first employed a response surface calculated between the reflectance values of a set of six certified photometric standards measured on both instruments, with all full range partial least square (PLS) regression models subsequently transferred. The next was correction of the spectra from the second instrument utilising the residual spectrum between the mean sample of the validation set measured on both instruments. Through this approach all PLS regression models and also a single multiple linear regression (MLR) model were transferred. As an outcome of this study guidelines are suggested for the transfer of NIR assays along with the criteria deemed necessary to conclusively prove transfer and justify any correction method utilised. The significant criteria were determined to be the paired t-test with both the UV reference assay data and the original NIR assay data, and comparison of the coefficient of multiple determinations. PMID- 12537381 TI - Light emitting diode excitation emission matrix fluorescence spectroscopy. AB - An excitation emission matrix (EEM) fluorescence instrument has been developed using a linear array of light emitting diodes (LED). The wavelengths covered extend from the upper UV through the visible spectrum: 370-640 nm. Using an LED array to excite fluorescence emission at multiple excitation wavelengths is a low cost alternative to an expensive high power lamp and imaging spectrograph. The LED-EEM system is a departure from other EEM spectroscopy systems in that LEDs often have broad excitation ranges which may overlap with neighboring channels. The LED array can be considered a hybrid between a spectroscopic and sensor system, as the broad LED excitation range produces a partially selective optical measurement. The instrument has been tested and characterized using fluorescent dyes: limits of detection (LOD) for 9,10-bis(phenylethynyl)-anthracene and rhodamine B were in the mid parts-per-trillion range; detection limits for the other compounds were in the low parts-per-billion range (< 5 ppb). The LED-EEMs were analyzed using parallel factor analysis (PARAFAC), which allowed the mathematical resolution of the individual contributions of the mono- and dianion fluorescein tautomers a priori. Correct identification and quantitation of six fluorescent dyes in two to six component mixtures (concentrations between 12.5 and 500 ppb) has been achieved with root mean squared errors of prediction (RMSEP) of less than 4.0 ppb for all components. PMID- 12537383 TI - Flow analysis-hydride generation-Fourier transform infrared spectrometry. A new analytical technique for the simultaneous determination of antimony, arsenic and tin. AB - The combination of flow analysis (FA), hydride generation (HG) and Fourier transform infrared (FTIR) spectrometry is proposed as a novel and powerful analytical technique for the individual and simultaneous determination of antimony, arsenic and tin in aqueous samples. The analytes were transformed into the volatile hydride form by on-line reaction with sodium tetrahydroborate in acidic medium. The gaseous analyte hydrides [M(n)H(m), (g)] generated, were transported by means of a carrier gas stream inside the IR gas cell and the corresponding FTIR spectrum was acquired in a continuous mode. The 1893, 1904 and 2115 cm(-1) bands of the SbH3, SnH4, and AsH3 were selected for the determination of antimony, tin and arsenic, respectively. The limit of detection (3sigma) obtained by using a short-path (10 cm) IR gas cell were 0.25, 0.30 and 1.2 mg l( 1) for the determination of antimony, tin and arsenic, respectively; while the precision (relative standard deviation, RSD, n 5) found from a standard solution containing 50 mg l(-1) of each element was, in all cases, less than 0.3%. However, the use of a long-path (7.25 m) IR gas cell improved the figures of merit (sensitivity, limits of detection and quantification) nearly 60-fold. The effect of the main experimental and instrumental variables, such as acidic media, sodium tetraborohydrate concentration, nitrogen flow rate, nominal resolution and the scan accumulation on the analytical signals of the antimony, tin and arsenic hydrides, were studied. Further, the potential of the proposed technique for the simultaneous determination of these elements was tested, analyzing synthetic samples containing different amounts of Sb, Sn and As. PMID- 12537382 TI - Detection of metalloproteins in human liver cytosol by synchrotron radiation X ray fluorescence combined with gel filtration chromatography and isoelectric focusing separation. AB - Synchrotron radiation X-ray fluorescence (SRXRF) spectroscopy is an advanced method of quantitative multielemental analysis with space resolution of several microm and sensitivities in the microg g(-1) range. It can be used for keeping track of trace elements after an electrophoretic separation of biological samples. In this paper, proteins in human liver cytosol were separated with gel filtration chromatography and thin layer isoelectric focusing (IEF). The contents of metal ions in protein bands were determined by SRXRF. The results showed that in the molecular weight (MW) range of 10 approximately 25 kDa, there were at least 2 Zn-containing bands with isoelectric point (pI) of 5 approximately 6 and 6.2 approximately 7, respectively and about 11 Fe-containing proteins with pI of 4.4, 4.6, 4.8, 5.0, 5.2, 5.3, 5.5, 5.6, 6.6, 6.8, and 7.2, respectively, present in human liver cytosol. The Zn-containing band with pI of 5-6 is the dominant species of zinc in this MW range. In addition, the Cu-containing bands with pI of 5.0 and below 4.8 were also detected. It is demonstrated that the procedure could be widely used in further investigations of the chemical species of trace elements in biological samples. PMID- 12537384 TI - Development of column-pretreatment chelating resins for matrix elimination/multi element determination by inductively coupled plasma-mass spectrometry. AB - Chelating resins, two kinds of iminodiacetate derivatives (IDA) of cross-linked chitosan (CCS) were synthesized and investigated for adsorption capacity, matrix elimination and collection/concentration of analytes by a column pretreatment in a multi-element ICP-MS determination method. The adsorption behavior of 54 elements at the 10 ng ml(-1) level on chitosan derivatives in a packed mini column was systematically examined. Almost 30 kinds of metal ions were recovered quantitatively at pH 5 with CCS-HP/IDA (cross-linked chitosan possessing N-2 hydroxypropyl iminodiacetic acid groups) column. Compared with available chitosan iminodiacetate resin, CHITOPEARL CI-03, the recovery of the metal ions such as Cu, Pb and La is satisfactory with CCS-IDA (cross-linked chitosan possessing N,N iminodiacetic acid groups) and CCS-HP/IDA using 2 M nitric acid as an eluent, which may be attributed to the difference of cross-linking and macroporous structure. Compared with Chelex-100, the adsorption efficiency is in the order: Chelex-100 > CCS-IDA > CCS-HP/IDA, especially in the chelating ability for alkaline earth metals. The resin with a longer spacer (CCS-HP/IDA) showed higher adsorption selectivity between heavy metal ions and alkaline earth metals at pH < 7. The separation efficiency of the major matrix cations in seawater (Na. K, Mg, Ca) has also been investigated, and matrix interference was negligible even in a seawater sample at pH 5 with CCS-HP/IDA. The recoveries of Mn at pH 5 with CCS HP/IDA or Chelex-100 were almost 100%. However, those of Mg with each resin were 4 or 98%, respectively. The adsorption capacities of synthesized CCS-HP/IDA for Cu(II), Pb(II) and La(III) were 0.90, 0.65 and 0.34 mmol g(-1), respectively. Therefore, the chelating chitosan resins developed are applicable to the pretreatment of trace amounts of elements in various kinds of water samples. PMID- 12537385 TI - Community animal health services for improving household wealth and health status of low-income farmers. AB - Community animal health services (CAHS) have been promoted since the 1970s by aid organizations in low-income countries to improve the health of animals. They are also thought to improve directly the wealth and health or livelihood of their owners. We have systematically searched for observational studies of basic preventive and curative animal health services provided by a community animal health worker in communities in low-income countries. The objective was to summarize the available research on the effects of CAHS on indicators for household wealth and health. Studies for inclusion were assessed independently according to predefined eligibility criteria, the methodological quality of the studies examined, and data extracted. Fourteen studies of varying methodological quality met the inclusion criteria. The results were unclear in four studies, and in the remaining studies it was difficult to summarize the results because they reported different outcomes. However, the studies report that CAHS has a considerable potential for improving human health and wealth as well as animal health and productivity. Given the amount of money and time invested in this area, these observations need to be confirmed in the context of a well-designed study using standard pragmatic outcomes. PMID- 12537386 TI - The control of rinderpest in Tanzania between 1997 and 1998. AB - In January 1997, Tanzania requested international assistance against rinderpest on the grounds that the virus had probably entered the country from southern Kenya. Over the next few months, a variety of attempts were made to determine the extent of the incursion by searching for serological and clinical evidence of the whereabouts of the virus. At the clinical level, these attempts were hampered by the low virulence of the strain, and at the serological level by the lack of a baseline against which contemporary interpretations could be made. Once it became apparent that neither surveillance tool was likely to produce a rapid result, an infected area was declared on common-sense grounds and emergency vaccination was initiated. The vaccination programme had two objectives, firstly to prevent any further entry across the international border, and secondly to contain and if possible eliminate rinderpest from those districts into which it had already entered. On the few occasions that clinical rinderpest was subsequently found, it was always within this provisional infected area. Emergency vaccination campaigns within the infected area ran from January to the end of March 1997 but were halted by the onset of the long rains. At this time, seromonitoring in two districts showed that viral persistence was still theoretically possible and therefore a second round of emergency vaccination was immediately organized. Further seromonitoring then indicated a large number of villages with population antibody prevalences of over 85%. These populations were considered to have been 'immunosterilized'. Although no clinical disease had been observed in them, it was decided to undertake additional vaccination in a group of districts to the south of the infected area. Serosurveillance indicated that rinderpest could have been present in a number of these districts prior to vaccination. Serosurveillance in 1998 suggested that numerous vaccinated animals had probably moved into districts outside the infected and additional vaccination areas, but did not rule out the continued presence of field infection. PMID- 12537387 TI - Growth and mortality in sheep and goats under high tsetse challenge in Kenya. AB - Trypanosomosis is a major impediment to livestock production and economic development in those areas of Africa where it is endemic. Although small ruminants appear to perform better than cattle in various agro-ecological zones, the importance of trypanosomosis has not been extensively investigated in these livestock. This study was designed to investigate the prevalence of trypanosomosis in sheep and goats in an endemic area and to evaluate the performance of different breeds under high tsetse challenge and the potential role of chemoprophylaxis in the control of the disease. The results showed that tsetse flies feed readily on small ruminants, and that these animals are susceptible to trypanosomosis. The Small East African goats acquired fewer infections than the Black Head Persian and Dorper sheep used in the study. In both sheep and goats, chemoprophylaxis with isometamidium chloride (Samorin, Rhone Merieux, Annecy, France) was protective, resulting in fewer infections and higher body weight gain. Trypanosomosis caused anaemia in both sheep and goats, and animals whose PCV fell below 15% rarely recovered, even with trypanocidal drug treatment. The peak transmission period was between 1 and 3 months after the peak tsetse fly density, which raises the possibility of effective strategic prophylaxis. PMID- 12537388 TI - Financial implications of rearing sheep and goats under natural trypanosomosis challenge at Galana ranch, Kenya. AB - A study to compare the profitability of rearing sheep and goats under natural trypanosomosis challenge was carried out on Galana ranch in south-eastern Kenya between July 1996 and October 1997. Seventy-nine male weaner sheep and 79 male weaner goats were monitored monthly for weight changes and fortnightly for trypanosomosis. The animals of each species were divided into two groups. Group 1 was an untreated control, while group 2 was treated with isometamidium chloride (Samorin) at 0.5 mg/kg body weight every 3 months. In both groups, trypanosome infections were detected by microscopy and treated with diminazene aceturate (Veriben), at 3.5 mg/kg body weight, when the packed cell volume reached 17% or below. The profitability of each drug regime was expressed as the marginal revenue over the cost of trypanosomosis (MOT). There were greater losses occasioned by trypanosomosis in sheep than in goats. Animals of both species on chemoprophylaxis gave higher MOT values than those that received chemotherapy on diagnosis. However, the MOT values for the chemoprophylactic regime were higher for sheep than for goats, suggesting that the greater weight gain by sheep more then compensated for the higher cost of maintaining them under high trypanosomosis challenge. Thus, a Galana rancher would be better off keeping sheep rather than goats, other things being equal. The marginal revenue per dose of Samorin was lower than that of Veriben for both species, suggesting that strategic use of Samorin timed to precede the peak incidence of trypanosomosis might be a better option to raise the overall profitability in sheep and goats. PMID- 12537389 TI - Characterization of indigenous fat-tailed and fat-rumped hair sheep in Kenya: diversity in blood proteins. AB - This paper reports the variation in five blood proteins from five populations of sheep found in Kenya. Blood samples were collected from a total of 309 adult sheep of both sexes in Kwale, Makueni and Kakamega districts for the fat-tailed sheep and in Isiolo district for the fat-rumped hair sheep. Fine-wooled Merino sheep were used in this study as the reference population. Transferrin, esterase A and esterase-C were polymorphic in all the populations investigated, while albumin was monomorphic for the S allele in the fat-tailed sheep and haemoglobin was fixed for the B allele in the Kwale, Makueni and Isiolo populations. Phylogenies derived from the pairwise genetic distance estimates showed a clear separation between the indigenous sheep populations and the exotic Merino. However, the topology of the former showed rather poor consistency with their morphological classification based on the localization of their fat deposits, namely fat-tailed or fat-rumped hair sheep. PMID- 12537390 TI - Milk production by agropastoral Red Sokoto goats in Nigeria. AB - The mild yield characteristics of smallholder agropastoral Fulani Red Sokoto goats were evaluated using records obtained from a total of 92 does in 6 herds, collected over a 2-year period. The results indicated that the total yield was 79.8 +/- 3.2 kg, the daily yield 0.66 +/- 0.03 kg, the peak yield 1.21 +/- 0.07 kg and the day of peak production 28.7 +/- 2.1 day of lactation. The milk yield characteristics were highly variable with a coefficient of variability of 38.6% to 69.8%. The characteristics were significantly (p < 0.01-0.05) influenced by herd, season, parity and litter size of the dam. Highly positive and significant (p < 0.01) correlations (r = 0.66-0.99) existed between the total yield and the average daily yield, peak yield, initial yield and day 120 yield. A negative and significant (p < 0.05) correlation (r = -0.33) was observed between the peak day and the initial yield. The non-homogeneity of the milk yield characteristics in Red Sokoto does suggests the possibility of improving their daily productivity by selection of high yielding does. Selection for higher initial milk yield may also improve other milk yield characteristics. PMID- 12537391 TI - Use of radioimmunoassay to measure progesterone levels during different reproductive stages in female Damascus goats. PMID- 12537392 TI - Artificial cells. General consideration. 1972 [classical article]. PMID- 12537393 TI - Artificial cells. Preparation. 1972 [classical article]. PMID- 12537394 TI - Artificial cells. Biophysical properties. 1972 [classical article]. PMID- 12537395 TI - Biological activity of artificial cells. 1972 [classical article]. PMID- 12537396 TI - Experimental enzyme therapy. 1972 [classical article]. PMID- 12537397 TI - Red blood cell substitutes. 1972 [classical article]. PMID- 12537398 TI - Artificial cells perfused by body fluids. 1972 [classical article]. PMID- 12537399 TI - Artificial cells containing detoxicants. 1972 [classical article]. PMID- 12537400 TI - Artificial cells for artificial organs. 1972 [classical article]. PMID- 12537401 TI - Artificial cells. General discussions and perspectives. 1972 [classical article]. PMID- 12537402 TI - Artificial cells. References to monograph articles. 1972 [classical article]. PMID- 12537403 TI - Cellular cardiomyoplasty: cell therapy for myocardial regeneration. AB - Cellular cardiomyoplasty (cell therapy for myocardial regeneration) targets the basic pathophysiology of heart failure, the irreversible loss of functional cardiomyocytes, and represents a novel means of augmenting cardiomyocyte number and contractile function of the failing heart. We discuss briefly the concept of myocardial regeneration and explore various cell types that may act as potential sources for myocardial tissue engineering. PMID- 12537404 TI - In vitro enzyme kinetics of microencapsulated tyrosinase. AB - In this study, we prepared microencapsulated tyrosinase and studied its enzyme kinetics and its stability at different pH and temperature. The Km for both free and microencapsulated tyrosinase is 465 microM. The Vmax for the microencapsulated tyrosinase is 49.02 mg/dl x min, whereas that of tyrosinase in free solution is 114.95 mg/dl x min. At the extreme pH of pH 2 and pH 10, free tyrosinase lost all its enzyme activity, whereas microencapsulated tyrosinase retained 14% of its original activity at pH 2 and 17% of its original activity at pH 10. Further in vitro studies were carried out to study the pH optimum of the microencapsulated enzyme at different pH values corresponding to those along the gastrointestinal tract. In temperature stability studies at 4 degrees C and 37 degrees C, higher enzyme activity remained in microencapsulated tyrosinase solution than that of free enzyme solution. Therefore, our results show that microencapsulated tyrosinase is significantly more stable than free tyrosinase. In subsequent studies, we show that daily oral administration of microencapsulated tyrosinase significantly lowers systemic tyrosinase after 3 days and maintains this low level as long as oral administration continues. When combined with one intravenous injection of polyhemoglobin-tyrosinase, the systemic tyrosine level can be lowered within one hour. PMID- 12537405 TI - Removal of middle molecules with sorbents. PMID- 12537406 TI - Is coronary stent deployment and remodeling affected by predilatation? An intravascular ultrasound randomized study Stenting with or without predilation: an IVUS study. AB - In this intravascular ultrasound (IVUS) randomized trial we compared a strategy of direct stenting (DS) without predilation (n = 30) vs. conventional stenting with predilation (SWP) (n = 30) in patients with suitable type A or B non calcified lesions in native vessels > or = 3 mm. Optimal deployment was achieved using angiographic criteria without interactive IVUS. The goal of our study was to determine whether stent expansion and coronary remodeling were similar. Maximal pressure inflation was comparable in the two groups (11.4 +/- 2.2 vs. 11.8 +/- 1.9 atm; NS). Stent deployment was obtained in all patients with complete apposition to the vessel wall. DS and SWP resulted in comparable lumen enlargement (5.4 +/- 2.5 vs. 5.5 +/- 2.1 mm2) with an identical mechanism: 66% of lumen enlargement was due to increased enlarged elastic membrane (EEM)-cross sectional area (CSA) (delta = 3.7 +/- 2.1 mm2 and delta = 2.4 +/- 6.8 mm2, respectively, p < 0.49) and 34% was due to a reduced P + M-CSA (delta = 0.02 +/- 6.9 mm2 and delta = 1.2 +/- 6.3 mm2, respectively, p < 0.50). We conclude that at the same maximal pressure inflation the mechanisms of stent expansion are similar in both DS and SWP groups. In this observational study, the IVUS data showed clearly under-expansion of stents in both groups in comparison with previously published CSA values (minimum stent CSA of 7.5 mm2). PMID- 12537407 TI - Time dependent changes of arterial distensibility induced by cholesterol and balloon injury in rabbits: an in vivo intravascular ultrasound study. AB - The aim of this study was to validate in vivo measurement of intravascular ultrasound (IVUS) for the analysis of structural and functional vessel wall alterations in a chronic animal model. Furthermore, the relation between functional and structural alteration of the vessel wall should be investigated. Fifteen cholesterol-fed rabbits (1%) and 15 control rabbits underwent balloon injury of the abdominal aorta. Immediately before and after balloon traumatization as well as 2 and 6 weeks later IVUS depiction of 10 aortal vessel segments was performed (n = 1,100 measurements). In vivo IVUS measurements and morphometric analysis of the neointimal area of same aortal segments showed a high correlation (n = 148, r = 0.844, p < 0.001). Plaque area determined by morphometry revealed larger areas than the evaluation by IVUS (0.162 +/- 0.138 vs. 0.130 +/- 0.126 mm2, p < 0.001). Before balloon traumatization, pulsatility of the aortal vessel segments was less in cholesterol-fed rabbits (0.067 vs. 0.090, p < 0.01) and neointimal index higher (0.003 vs. 0). Investigation using IVUS 2 and 6 weeks after balloon traumatization demonstrated a continuous loss of arterial distensibility and an increase of neointimal index, being more pronounced in the cholesterol-fed group. As demonstrated by IVUS the loss of distensibility preceded the atherosclerotic alterations. Our investigation suggests using IVUS in this animal model is a reliable setting for long-term investigation of characteristics of the vessel wall. We could demonstrate that altered function of the vessel wall precedes the structural atherosclerotic vessel wall alterations. PMID- 12537408 TI - Contrast agents provide a faster learning curve in dipyridamole stress echocardiography. AB - AIM: Interobserver variability is an important limitation of the stress echocardiography and depends on the echocardiographer training. Our aim was to evaluate if the use of contrast agents during dipyridamole stress echocardiography would improve the agreement between an experienced and a non experienced observer in stress echo and therefore if contrast would affect the learning period of dypyridamole stress echo. METHODS AND RESULTS: Two independent observers without knowledge of any patient data interpreted all stress studies. One observer was an experienced one and the other had experience in echocardiography but not in stress echo. Two observers analysed 87 non-selected and consecutive studies. Out of the 87 studies, 46 were performed without contrast administration, whereas i.v. contrast (2.5 g Levovist by two bolus at rest and at peak stress) was administered in 41. In all cases, second harmonic imaging and stress digitalisation pack was used. The agreement between observers showed a kappa index of 0.58 and 0.83 without and with contrast administration, respectively. CONCLUSIONS: The use of contrast agents provides a better agreement in the evaluation of stress echo between an experienced and a non-experienced observer in stress echo. Adding routinely contrast agents could probably reduce the number of exams required for the necessary learning curve in stress echocardiography. PMID- 12537409 TI - Elongation as a new shape index for the left ventricle. AB - OBJECTIVES: This study was done to quantify the shape of the left ventricle (LV). It was proposed that the shape of the LV is intimately related to its performance and that its elongation (ELO) is a sensitive measure of this performance. The performance was tested against classical cardiovascular parameters. METHODS: Using echocardiography and Simpson's rule, the endocardial surface area of the LV was calculated noninvasively with a simple experimental-mathematical model at enddiastole and endsystole. ELO as shape index was derived from the endocardial surface area of the LV with a simple formula. The endocardial surface area of the LV and ELO were determined in volunteers, in patients with mild heart failure and in patients with severe heart failure. RESULTS: The normal value of endocardial surface area of LV at enddiastole is 138.3 cm2 while the normal value at endsystole is 99 cm2. The endocardial surface area of the LV is significantly bigger in patients with mild heart failure than in volunteers (p < 0.01) while the parameters ELO, ejection fraction and Doppler measurements are similar. The normal values of ELO at diastole and systole are 12 and 25 respectively. The value of ELO at endsystole is lower only in patients with severe heart failure. This means a more spherical shape and poor systolic function of the LV. CONCLUSION: ELO is usefull as quantitative and qualitative index of left ventricular shape. ELO could be integrated and applied with new diagnostic tools such three-dimensional and contrast echocardiography. PMID- 12537410 TI - Widespread thrombosis associated with recurrent intracardiac masses in a patient with Behcet's disease. PMID- 12537411 TI - Ebstein's anomaly in siblings: an original observation. AB - Siblings, a 26-year old male and a 31-year old female, were found do have Ebstein's anomaly. Ebstein's anomaly is characterized by a downward displacement of the tricuspid valve into the right ventricle due to anomalous attachment of the tricuspid leaflets. Echocardiography is the method of choice to diagnose Ebstein's anomaly on its own or in association with other heart defects. Complications such as right ventricular failure, infective endocarditis, and paradoxical embolism can occur. Ebstein's anomaly diagnosed in adult life is a benign and stable disease, particularly if the patient is asymptomatic. These cases may represent a familial form of Ebstein's anomaly. PMID- 12537412 TI - Ebstein's anomaly in adults. PMID- 12537414 TI - Clinical experience with coronary sinus catheterization in minimally invasive aortic valve surgery under transesophageal echocardiography guidance. AB - We aimed to show conventional coronary sinus (CS) catheter could be used with transesophageal echocardiography (TEE) guidance through the limited surgical field in aortic valve surgery with 'J' sternotomy. This method was performed in 14 patients and completed successfully in 12. We believe that in minimally invasive aortic valve surgery, the insertion of the conventional retrograde cardioplegia catheter to the CS with routine way may not be possible but application of TEE guidance is cost-effective and easily applicable method without significant complications. PMID- 12537413 TI - Clinical application in routine practice of the proximal flow convergence method to calculate the mitral surface area in mitral valve stenosis. AB - BACKGROUND: Two-dimensional (2D) echocardiography planimetry, the Doppler pression half-time (PHT), and the continuity equation methods were used to estimate mitral valve area (MVA) in patients with mitral stenosis (MS). Recently, the proximal isovelocity surface area (PISA) method has been shown to be accurate for calculating MVA. The purpose of this study is (1) to compare in a large non selected population the accuracy of the PISA and planimetry methods for echocardiographic estimation of MVA; (2) to determine the effect of atrial fibrillation (AF), Wilkins score, associated mitral regurgitation (MR), aortic regurgitation (AR), and of commissural calcifications on the accuracy of the PISA method. METHODS: One hundred and eight consecutive patients with rheumatic MS were studied (76 females and 32 males; mean age: 36 +/- 12 years); 64 were in sinus rhythm; 51 had associated MR and 46 had AR. By the PISA method. MVA was calculated assuming a uniform radius flow convergence region along a hemispherical surface. RESULTS: The mean value of 2D MVA was 1.32 +/- 0.59 cm2 (0.4-3.1 cm2) and that of PISA MVA 1.33 +/- 0.62 cm2 (0.38-3 cm2). MVA calculated using the PISA method correlated well with 2D MVA (r = 0.93, y = 0.97x + 0.04, p < 0.0001, SEE = 0.21 cm2). The correlation was also good in patients with AF (r = 0.93, y = 0.99x + 0.03, p < 0.0001, SEE = 0.21 cm2), with MR (r = 0.94, y = 1.0 14x + 0.003, p < 0.0001, SEE = 0.19 cm2), with AR (r = 0.93, y = 0.90x + 0.11, p < 0.0001, SEE = 0.2 cm2), when Wilkins score was >8 (r = 0.92, = 0.96x + 0.06, p < 0.0001, SEE = 0.19 cm2), and in patients with commissural calcifications (r = 0.90, y = 0.88x + 0.009, p < 0.0001, SEE = 0.20 cm2). CONCLUSION: Our study shows that in routine practice, MVA calculated by the PISA method correlated well with the area obtained by planimetry even in the presence of commissural calcifications, associated MR, AR, AF and of high Wilkins score. Therefore, the PISA method provides a reliable measurement of the MVA in MS under different anatomic and clinical conditions and may be a useful alternative method for calculating MVA. PMID- 12537415 TI - Detection of the sinus node artery using electron beam computed tomography of the heart. AB - BACKGROUND: Stenosis or obstruction of the sinus node artery (SNA) of the heart can cause cardiac arrhythmia and even sudden death. The purpose of this study was to evaluate the detectability of the SNA using electron beam computed tomography (EBCT). MATERIAL AND METHODS: Eighty patients (mean age: 61 +/- 10 years, range: 31-80 years) were examined with ECG-triggered EBCT (exposure time: 100 ms, slice thickness: 1.5 mm), using two different detector systems (n = 40; 9.5 line pairs/cm [Lp/cm]; n = 40; 7 Lp/ cm) after intravenous contrast agent administration. Twenty-four out of 80 (30%) patients had supraventricular arrhythmia. RESULT: The SNA was detected in 60/80 (75%) of all patients with an average length of 16 +/- 7 mm (3-35.2 mm). One additional branch of the SNA was detectable in 17/80 patients (21%), two branches in 6/80 (8%). There was no significant difference between the two used detector systems (p < 0.01) or between patients with or without supraventricular arrhythmia (71 vs. 77%; p = ns). CONCLUSION: EBCT allows to detect small coronary arteries such as the SNA of the heart. PMID- 12537416 TI - Left cervical aortic arch with aneurysm and obstruction: three-dimensional computed tomographic angiography and magnetic resonance angiographic appearance. AB - Cervical arch is a rare congenital anomaly presumed to result from persistence of the third aortic arch and regression of the normal fourth arch. Rather rare is cervical aortic arch associated with aneurysm and obstruction, with eight known cases reported. Definitive diagnosis with a noninvasive imaging modality is desirable and very important to prevent the need for disaster intervention. We present two cases of a pulsatile mass in the left supraclavicular region. Three dimensional computed tomographic angiography and magnetic resonance angiography clearly showed a left-sided cervical aortic arch (Haughton type D) with arch aneurysm and coarctation (pseudocoarctation). PMID- 12537417 TI - Non-invasive magnetic resonance imaging of coronary bypass grafts. comparison of the haste- and navigator techniques with conventional coronary angiography. AB - OBJECTIVE: The aim was the study was to evaluate the patency of coronary artery bypass grafts with the 3D Navigator and the Haste magnetic resonance imaging techniques. MATERIAL AND METHODS: Thirty-four patients with 82 grafts (18 internal mammarian artery and 64 saphenous venous grafts) were examined at 1.5 T using a phased arrray body coil. A 2D T-2 weighted turbo spin echo sequence (Haste) and a 3D-angiography sequence in navigator technique were used. Reference method was the conventional coronary angiography that was performed within 7 days. RESULTS: With the HASTE sequence 54 of 56 patent and 12 of the 16 occluded grafts were recognized, the sensitivity and specificity was 96 and 75%. The distal anastomosis could be assessed in 28 of 76 cases. With the Haste sequence 54 of the 56 patent and 12 of the 14 occluded grafts were detected correctly. Eight patent and two occluded grafts were judged as not assessable. The sensitivity and specificity was 96 and 85%. Sixty-six percent (50/76) of the distal anastomosis were identified correctly. With both sequences none of the bypass stenosis could be detected. CONCLUSIONS: The patency of coronary bypass grafts can be evaluated noninvasively with a sensitivity of 96% and a specificity of 76%. No inference can be made with respect to the nature of bypass graft stenosis. Better results can be expected with the development of a blood pool contrast medium, an improvement of the spatial resolution and in the combination of flow measurements. PMID- 12537418 TI - Magnetic resonance imaging for the non-invasive detection of stenosis in coronary artery bypass grafts: clinical reality? PMID- 12537419 TI - Quantification of the predominant monomeric catechins in baking chocolate standard reference material by LC/APCI-MS. AB - Catechins are polyphenolic plant compounds (flavonoids) that may offer significant health benefits to humans. These benefits stem largely from their anticarcinogenic, antioxidant, and antimutagenic properties. Recent epidemiological studies suggest that the consumption of flavonoid-containing foods is associated with reduced risk of cardiovascular disease. Chocolate is a natural cocoa bean-based product that reportedly contains high levels of monomeric, oligomeric, and polymeric catechins. We have applied solid-liquid extraction and liquid chromatography coupled with atmospheric pressure chemical ionization-mass spectrometry to the identification and determination of the predominant monomeric catechins, (+)-catechin and (-)-epicatechin, in a baking chocolate Standard Reference Material (NIST Standard Reference Material 2384). (+)-Catechin and (-)-epicatechin are detected and quantified in chocolate extracts on the basis of selected-ion monitoring of their protonated [M + H](+) molecular ions. Tryptophan methyl ester is used as an internal standard. The developed method has the capacity to accurately quantify as little as 0.1 microg/mL (0.01 mg of catechin/g of chocolate) of either catechin in chocolate extracts, and the method has additionally been used to certify (+)-catechin and ( )-epicatechin levels in the baking chocolate Standard Reference Material. This is the first reported use of liquid chromatography/mass spectrometry for the quantitative determination of monomeric catechins in chocolate and the only report certifying monomeric catechin levels in a food-based Standard Reference Material. PMID- 12537420 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of heteropolyflavan-3-ols and glucosylated heteropolyflavans in sorghum [Sorghum bicolor (L.) Moench]. AB - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to characterize the structural diversity of polyflavans in Ruby Red sorghum [Sorghum bicolor (L.) Moench]. Deionization of the polyflavan fractions with Dowex 50 x 8-400 cation-exchange resin and subsequent addition of cesium trifluoroacetate ((133)Cs) allowed the detection of exclusively [M + Cs](+) ions. MALDI-TOF MS of the polyflavans that eluate from Sephadex LH-20 columns with methanol and acetone detected a series of masses corresponding to heteropolyflavan-3-ols differing in degree of hydroxylation and nature of the interflavan bond (A-type and B-type). MALDI-TOF MS of the Sephadex ethanol/methanol (v/v) eluate revealed a series of masses corresponding to heteropolyflavan-5-O-beta-glucosides that vary in the extent of hydroxylation and contain a flavanone (eriodictyol or eriodictyol-5-O-beta-glucoside) as the terminal unit. The combination of liquid chromatographic separation and MALDI-TOF MS to characterize sorghum polyflavans indicates that the structural heterogeneity is much greater than previously described. PMID- 12537421 TI - Native fluorescence from juvenile stages of common food storage insects. AB - Insect infestation of stored foods has significant economic and health consequences; the development of novel methods of detection thus presents considerable opportunities. The fluorescence from nine species of storage insects (beetles and moths) was studied; the juvenile stages of all nine species exhibited fluorescence under long-wave (365 nm) UV light; none of the adult insects emit fluorescence, so the fluorophore(s) might be a compound(s) associated with the unsclerotized cuticle. The spectra of larval stages of Ephestia kuehniella, Oryzaephilus surinamensis, Corcyra cepahlonica, Tribolium castaneum, and Tribolium confusum exhibited excitation maxima in the range from 345 to 350 nm and emission maxima in the range from 421 to 427 nm, suggesting that fluorescence arises from a common chromophore; similarities in fluorescence properties implicate one of the many pteridine ring-containing compounds (pterins) commonly found in insects. Larvae and even eggs were readily imaged on foods using fluorescence under 365-nm excitation. Fluorescence thus appears to be ubiquitous in immature food storage insects, and fluorescence detection may be useful as a general method to detect insects in foods and agricultural commodities during storage or processing. PMID- 12537422 TI - Assessment of screening methods for the identification of genetically modified potatoes in raw materials and finished products. AB - Qualitative polymerase chain reaction methods for the detection of genetically modified potatoes have been investigated that can be used for screening purposes and identification of insect-resistant and virus-resistant potatoes in food. The presence of the nos terminator from Agrobacterium tumefaciens and the antibiotic marker gene nptII (neomycin-phosphotransferase II) was demonstrated in three commercialized Bt-potato lines (Monsanto Co., St. Louis, MO, USA) and one noncommercial GM-potato product (high amylopectin starch, AVEBE, Veendam, The Netherlands) and allows for general screening in foods. For further identification, specific primers for the FMV promoter derived from the figwort mosaic virus, the CryIIIA gene (delta-endotoxin from Bacillus thuringiensis subsp. tenebrionis), potato leafroll virus replicase gene, and the potato virus Y coat protein gene, were designed. The methods described were successfully applied to processed potato raw materials (dehydrated potato powders and flakes), starch samples, and finished products. PMID- 12537423 TI - A simple and sensitive liquid chromatography-mass spectrometry confirmatory method for analyzing sulfonamide antibacterials in milk and egg. AB - A simple and specific method able to identify and quantify traces of 14 sulfonamide antibacterials (SAs) in milk and eggs is presented. This method uses a single solid-phase extraction (SPE) cartridge for simultaneous extraction and purification of SAs in the above matrices. Milk and egg samples are passed through a Carbograph 4 sorption cartridge. After analyte desorption, an aliquot of the final extract is injected into a liquid chromatography-mass spectrometry (LC-MS) instrument equipped with an electrospray ion source (ESI) and a single quadrupole. MS data acquisition is performed in the positive-ion mode and by a time-scheduled multiple-ion selected ion monitoring program. Compared to two published methods, the present protocol extracted larger amounts of SAs from both milk and egg and decreased the analysis time by a factor of 3 with milk samples and by a factor of 2 with egg samples. Recovery of SAs in milk at the 5 ppb level ranged between 76 and 112% with relative standard deviations (RSDs) of caffeic > ferulic > p-coumaric acid. The avenanthramides derived from 2 were usually slightly more active than those derived from 1. All avenanthramides inhibited azo initiated peroxidation of linoleic acid. 1c and 1s were initially the most effective compounds. The relative order of antioxidant activities was slightly different for the DPPH and the linoleic acid assays run in methanol and chlorobenzene, respectively. PMID- 12537429 TI - Analysis of antioxidative phenolic compounds in artichoke (Cynara scolymus L.). AB - Artichoke leaf is an herbal medicine known for a long time. A systematic antioxidant activity-directed fractionation procedure was used to purify antioxidative components from the aqueous methanol extractions of artichoke heads and leaves in this study. Seven active polyphenolic compounds were purified from artichoke, and structural elucidation of each was achieved using MS and NMR. Two of these compounds, apigenin-7-rutinoside and narirutin, were found to be unique to artichoke heads, this represents the first report of these compounds in the edible portion of this plant. The contents of these antioxidants and total phenols in dried artichoke samples from leaves and immature and mature heads of three varieties, Imperial Star, Green Globe, and Violet, were then analyzed and compared by colorimetric and validated HPLC methods. Significant differences by variety and plant organ were observed. PMID- 12537430 TI - Antioxidant activity of apple peels. AB - Consumption of fruits and vegetables has been shown to be effective in the prevention of chronic diseases. These benefits are often attributed to the high antioxidant content of some plant foods. Apples are commonly eaten and are large contributors of phenolic compounds in European and North American diets. The peels of apples, in particular, are high in phenolics. During applesauce and canned apple manufacture, the antioxidant-rich peels of apples are discarded. To determine if a useful source of antioxidants is being wasted, the phytochemical content, antioxidant activity, and antiproliferative activity of the peels of four varieties of apples (Rome Beauty, Idared, Cortland, and Golden Delicious) commonly used in applesauce production in New York state were investigated. The values of the peels were compared to those of the flesh and flesh + peel components of the apples. Within each variety, the total phenolic and flavonoid contents were highest in the peels, followed by the flesh + peel and the flesh. Idared and Rome Beauty apple peels had the highest total phenolic contents (588.9 +/- 83.2 and 500.2 +/- 13.7 mg of gallic acid equivalents/100 g of peels, respectively). Rome Beauty and Idared peels were also highest in flavonoids (306.1 +/- 6.7 and 303.2 +/- 41.5 mg of catechin equivalents/100 g of peels, respectively). Of the four varieties, Idared apple peels had the most anthocyanins, with 26.8 +/- 6.5 mg of cyanidin 3-glucoside equivalents/100 g of peels. The peels all had significantly higher total antioxidant activities than the flesh + peel and flesh of the apple varieties examined. Idared peels had the greatest antioxidant activity (312.2 +/- 9.8 micromol of vitamin C equivalents/g of peels). Apple peels were also shown to more effectively inhibit the growth of HepG(2) human liver cancer cells than the other apple components. Rome Beauty apple peels showed the most bioactivity, inhibiting cell proliferation by 50% at the low concentration of 12.4 +/- 0.4 mg of peels/mL. The high content of phenolic compounds, antioxidant activity, and antiproliferative activity of apple peels indicate that they may impart health benefits when consumed and should be regarded as a valuable source of antioxidants. PMID- 12537431 TI - Quantitative kinetic analysis of hydrogen transfer reactions from dietary polyphenols to the DPPH radical. AB - Diphenylpicrylhydrazyl (DPPH) is widely used for quickly assessing the ability of polyphenols to transfer labile H atoms to radicals, a likely mechanism of antioxidant protection. This popular test generally pays no attention to the kinetics of H atom transfer, which however could be even more important than the total H-atom-donating capacities (stoichiometry, EC50) typically evaluated. In the present work, a series of dietary polyphenols belonging to the most representative families (flavonols from onion, flavanol monomers and oligomers from barley, and caffeic acid and caffeoyl esters from artichoke and endive) are characterized not only by their total stoichiometries (n(tot)) but also by their rate constants of first H atom abstraction by DPPH (k(1)), deduced from the kinetic analysis of the decay of the DPPH visible band following addition of the antioxidant. The mildly reactive DPPH radical allows a good discrimation between polyphenols, as demonstrated by the relatively large ranges of k(1) (ca. 400-5000 M(-)(1) s(-)(1)) and n(tot) (ca. 1-5) values typically measured with antioxidants having a single polyphenolic nucleus. With antioxidants displaying more than one polyphenolic nucleus (procyanidin oligomers, dicaffeoyl esters), the kinetic analysis makes it possible to demonstrate significant differences in reactivity between the subunits (two distinct k(1) values whose ratio lies in the range 3 10) and nonadditive stoichiometries. PMID- 12537432 TI - Enhancing quality and oxidative stability of aged fried food with gamma tocopherol. AB - To determine the effects of gamma-tocopherol on the stability of fried food, potato chips were fried in triolein with 0, 100, or 400 ppm gamma-tocopherol. Potato chips, sampled at 1, 3, and 6 h of frying time, were aged for 0, 2, and 4 days at 60 degrees C and then evaluated for odor attributes by sensory analysis and for volatile compounds by purge-and-trap gas chromatography-mass spectrometry. Oil sampled after 1, 3, and 6 h of frying time from the fryer was evaluated for total polar compounds and retention of gamma-tocopherol. Oil extracted from the potato chips was also analyzed for residual gamma-tocopherol. gamma-Tocopherol disappeared rapidly, with only slight amounts of the original 100 ppm level detectable after the triolein was used for frying. gamma-Tocopherol significantly inhibited polar compound production in the triolein. Results showed that gamma-tocopherol inhibited the oxidation of the fried food even when only very low levels of retained gamma-tocopherol were present in the frying oil or potato chips. Nonanal formation was inhibited by gamma-tocopherol in aged potato chips. Odor analysis of the aged potato chips showed that samples with 0 ppm gamma-tocopherol had a rancid odor after being aged for 4 days. Potato chips with 400 ppm gamma-tocopherol had no rancid odors; however, as the level of gamma tocopherol decreased in the triolein and in the potato chips, a weak plastic odor characteristic of oxidized triolein was detected. PMID- 12537433 TI - Antioxidant activity of anthocyanins and their aglycons. AB - The antioxidant activity of the six common anthocyanidins, pelargonidin, cyanidin, delphinidin, peonidin, petunidin, and malvidin, and their glycosidic forms was evaluated in three lipid-containing models [human low-density lipoprotein (LDL) and bulk and emulsified methyl linoleate]. In addition, the radical scavenging activity of the compounds against the 2,2-diphenyl-1 picrylhydrazyl (DPPH) radical was studied. Most anthocyanins and their aglycons acted as strong antioxidants in emulsion and LDL. Many compounds showed an activity comparable to the well-known antioxidants alpha-tocopherol, Trolox, catechin, and quercetin. In bulk methyl linoleate, anthocyanins and anthocyanidins possessed only a weak antioxidant activity or even oxidation promoting activity. Depending on the anthocyanidin, different glycosylation patterns either enhanced or diminished the antioxidant power. For the most part, the activities of the glycosides and the aglycons did not differ remarkably in emulsion. In LDL the aglycons showed in general higher activities than the glycosides. In bulk oil, to the contrary, the glycosides were more effective than the aglycons. PMID- 12537434 TI - Oxygen permeability of films made from CO2-precipitated casein and modified casein. AB - Oxygen permeabilities (OP) of CO(2)-casein (CO(2)CN), calcium caseinate (CaCN), and acylated casein (AcCN) films were determined as functions of % relative humidity (% RH), temperature, and plasticizer type. Tensile properties and water vapor permeabilities (WVP) were also measured. Plasticizers were glycerol (GLY) or a 3:1 ratio of GLY:poly(propylene glycol) (PPG), a hydrophobic plasticizer. OP of the CO(2)CN:GLY film was almost twice that of films containing either plasticizer at 35% RH, but its OP approached that of the other films at 70% RH. OP and WVP of films plasticized with GLY were greater than that for films plasticized with PPG. Plasticizer type had little impact on the tensile strength of CO(2)CN films while tensile strength of CaCN-GLY:PPG (3:1) films approximately doubled. Results show that structural dissimilarities in the films contribute to differences in OP only under conditions of low RH where the plasticizing effects of water are not significant. PMID- 12537435 TI - Effects of cultivar and processing method on the contents of catechins and procyanidins in grape juice. AB - The aim of the presented work was to study the effect of pressing method, pasteurization, cultivar, and vintage on the content of (+)-catechin, (-) epicatechin, and nine procyanidins in grape juice. The results showed that the concentration of these flavan-3-ols in the juice was influenced, in decreasing order of importance, by pressing method, cultivar, pasteurization, and vintage. Cold pressing without maceration was the least and hot pressing after maceration at 60 degrees C for 60 min the most effective method for extracting the flavan-3 ols. Pasteurization increased the concentration of catechins in cold-pressed juices, but it decreased concentrations in hot-pressed juices. The concentration of most procyanidins was increased by pasteurization. Among the white cultivars, Seyval and Niagara were highest in procyanidins and Elvira and Chardonnay were highest in catechins. Vincent, Foch, and Baco were the red cultivars highest in catechins, and Vincent also had the highest content of procyanidins. PMID- 12537436 TI - Vitamin C, provitamin A carotenoids, and other carotenoids in high-pressurized orange juice during refrigerated storage. AB - Vitamin C, provitamin A carotenoids, and other carotenoids were measured in freshly squeezed juices from oranges (Citrus sinensis L. var. Valencia late) that were subjected to high-pressure (HP) treatment. Also, the stability of these compounds was studied during refrigerated storage at 4 degrees C. HP treatment is an alternative to heat preservation methods for foods; therefore, it is essential to assess the impact of HP on bioactive compounds. Several processes that combine HP treatment with heat treatment for various time periods were assayed: T0, fresh juice (without treatment); T1, 100 MPa/60 degrees C/5 min; T2, 350 MPa/30 degrees C/2.5 min; T3, 400 MPa/40 degrees C/1 min. Fresh and treated samples were kept refrigerated (4 degrees C) over 10 days. After application of HP and during the refrigeration period, the qualitative and quantitative determination of vitamin C, provitamin A carotenoids (beta- and alpha-carotene; beta- and alpha cryptoxanthin), and the xanthophylls zeaxanthin and lutein was achieved by high performance liquid chromatography. T1 and T3 juices showed a decrease in ascorbic acid and total vitamin C just after HP treatment (D0) compared with T0 juices. On the contrary, T2 juices, just after HP treatment (D0), had the same levels of both compounds compared to untreated juices. T1, T2, and T3 treatments led to an increase in the extraction of carotenoids and provitamin A carotenoids. Total carotenoid content after the 10-day refrigerated storage period resulted in no significant quantitative changes in T1 juices, whereas in T2 and T3 juices small losses were found at the end of the storage period (20.56 and 9.16%, respectively). These losses could be influenced by the depleted protection of vitamin C toward carotenoid oxidation during the same period. A similar trend was found in provitamin A carotenoids for the different treated juices. PMID- 12537437 TI - Storage of immobilized yeast cells for use in wine-making at ambient temperature. AB - A comparative study of the storage and reuse of immobilized yeast cells on apple pieces, kissiris, and gamma-alumina was carried out. The immobilized biocatalysts were allowed to remain in the fermented alcoholic liquid after the end of each fermentation batch for extended periods at 30 degrees C before reactivation in batch fermentation for wine-making. The results showed that the biocatalysts were able to reactivate and ferment after successively increased periods of storage compared to free cell systems both on glucose medium and on grape must. In glucose medium, apple-, kissiris-, and gamma-alumina-supported biocatalysts reactivated after 120, 80, and 83 days, respectively. Possible storage periods for grape must were lower but remained high. Immobilized yeast biocatalyst on apple pieces produced wines with an improved volatiles composition compared to kissiris- and gamma-alumina-supported biocatalysts. There were no significant negative effects on the fermentation activity and volatile byproduct composition. PMID- 12537438 TI - Effects of processing conditions on the physical and chemical properties of buckwheat grit cakes. AB - Buckwheat grit cakes were prepared with a rice cake machine using the following independent variables: tempering moisture contents (15, 17, and 19%, wb), heating temperatures (240, 246, 252, and 258 degrees C), and heating times (5, 6, 7, and 8 s). Higher moisture, higher heating temperature, or longer heating time produced cakes with a higher cake specific volume. Cakes became lighter in color at a lower heating temperature or a shorter heating time. The hardest cake was produced at 252 degrees C for 5 s at 19% moisture content. The percent weight loss after tumbling decreased with increasing heating times and heating temperatures. Increased heating time resulted in more expanded products. The average rutin content decreased as the heating temperature or heating time increased. These results suggest that processing conditions, including tempering moisture, heating temperature, and heating time, significantly influenced physical and chemical qualities of buckwheat grit cakes such as specific volume, hardness, integrity, color, internal structure, and rutin content. PMID- 12537439 TI - Changes in phenolic composition and antioxidant activity of virgin olive oil during frying. AB - The concentration of hydroxytyrosol (3,4-DHPEA) and its secoiridoid derivatives (3,4-DHPEA-EDA and 3,4-DHPEA-EA) in virgin olive oil decreased rapidly when the oil was repeatedly used for preparing french fries in deep-fat frying operations. At the end of the first frying process (10 min at 180 degrees C), the concentration of the dihydroxyphenol components was reduced to 50-60% of the original value, and after six frying operations only about 10% of the initial components remained. However, tyrosol (p-HPEA) and its derivatives (p-HPEA-EDA and p-HPEA-EA) in the oil were much more stable during 12 frying operations. The reduction in their original concentration was much smaller than that for hydroxytyrosol and its derivatives and showed a roughly linear relationship with the number of frying operations. The antioxidant activity of the phenolic extract measured using the DPPH test rapidly diminished during the first six frying processes, from a total antioxidant activity higher than 740 micromol of Trolox/kg down to less than 250 micromol/kg. On the other hand, the concentration of polar compounds, oxidized triacylglycerol monomers (oxTGs), dimeric TGs, and polymerized TGs rapidly increased from the sixth frying operation onward, when the antioxidant activity of the phenolic extract was very low, and as a consequence the oil was much more susceptible to oxidation. The loss of antioxidant activity in the phenolic fraction due to deep-fat frying was confirmed by the storage oil and oil-in-water emulsions containing added extracts from olive oil used for 12 frying operations. PMID- 12537440 TI - Accelerated degradation of methyl iodide by agrochemicals. AB - The fumigant methyl iodide (MeI, iodomethane) is considered a promising alternative to methyl bromide (MeBr) for soil-borne pest control in high-cash value crops. However, the high vapor pressure of MeI results in emissions of a significant proportion of the applied mass into the ambient air, and this may lead to pollution of the environment. Integrating the application of certain agrochemicals with soil fumigation provides a novel approach to reduce excessive fumigant emissions. This study investigated the potential for several agrochemicals that are commonly used in farming operations, including fertilizers and nitrification inhibitors, to transform MeI in aqueous solution. The pseudo first-order hydrolysis half-life (t(1/2)) of MeI was approximately 108 d, while the transformation of MeI in aqueous solutions containing selected agrochemicals was more rapid, with t(1/2) < 100 d (t(1/2) < 0.5 d in some solutions containing nitrification inhibitors). The influence of these agrochemicals on the rate of MeI degradation in soil was also determined. Adsorption to soil apparently reduced the availability of some nitrification inhibitors in the soil aqueous phase and lowered the degradation rate in soil. In contrast, addition of the nitrification inhibitors thiourea and allylthiourea to soil significantly accelerated the degradation of MeI, possibly due to soil surface catalysis. The t(1/2) of MeI was <20 h in thiourea- and allylthiourea-amended soil, considerably less than that in unamended soil (t(1/2) > 300 h). PMID- 12537441 TI - Shikimate accumulates in both glyphosate-sensitive and glyphosate-resistant horseweed (Conyza canadensis L. Cronq.). AB - Horseweed (Conyza canadensis) is a cosmopolitan weed that commonly grows throughout North America. Horseweed that is not completely controlled by normal applications of glyphosate has been reported in western Tennessee. This research had three objectives: (1) to develop and validate an analytical procedure for the quantitative determination of shikimate, an important indicator of glyphosate activity in plants; (2) to confirm resistance to glyphosate in a horseweed population; and (3) to examine the accumulation of shikimate in both glyphosate resistant and glyphosate-susceptible horseweed plants. The analytical procedure to determine shikimate used extraction with 1 M HCl for 24 h, followed by liquid chromatography using photodiode array detection, and shikimate recoveries were >or=82%. Glyphosate applications of both 0.84 kg ae/ha (the standard application rate) and 3.8 kg ae/ha to susceptible plants caused complete plant death. The same glyphosate applications to putative resistant populations caused less than 15% growth reduction as determined by visual evaluations, and fresh weights of these resistant plants 17 days after glyphosate treatment (DAT) were reduced an average of 45% in one population and were not affected in a different population. This direct comparison conclusively confirms that horseweed plants collected in western Tennessee in 2002 are resistant to 4 times the normal application dosage of glyphosate. The glyphosate-resistant horseweed biotypes still exhibited some herbicidal effects from the glyphosate, such as yellowing in the most actively growing, apical shoot meristems. The yellowing in the shoot apexes was transitory, and the plants recovered from this damage. Shikimate concentrations in all untreated horseweed plants were less than 100 microg/g, which was significantly less than that in all plants which had been treated with 0.84 kg ae/ha of glyphosate. Unexpectedly, shikimate accumulated (>1000 microg/g) in both resistant populations and the susceptible population. However, there were differences in shikimate accumulation patterns between resistant and susceptible horseweed biotypes. Shikimate concentrations in resistant populations declined about 40% from 2 to 4 DAT, while shikimate concentrations in the susceptible horseweed plants increased about 35% from 2 to 4 DAT. The confirmed resistance of a widespread weed implies that alternative control strategies for glyphosate resistant horseweed will be needed in those no-tillage production systems where it commonly occurs. PMID- 12537442 TI - Leaching and binding of fumonisins in soil microcosms. AB - Fumonisin B(1) (FB(1)) is a water-soluble mycotoxin produced by Fusarium verticillioides. Our research objectives were to determine the leaching of FB(1) through soils and FB(1) binding in soil. Leachate columns were used to determine the movement of FB(1) through soil. FB(1)-contaminated corn screenings or water extracts containing FB(1) were placed on the surface of soil columns. In 100% sand columns, FB(1) leaching was only slightly retarded, whereas at 50%, 75%, and 100% Cecil sandy loam, approximately 60%, 50%, and 20% of the FB(1) was recovered in the column leachate, respectively. The FB(1) retained on the 100% Cecil sandy loam column was tightly bound. However, approximately 75% of the bound FB(1) was released with 5% formic acid and 5% formic acid/acetonitrile (1:1), indicating that the nature of the interaction was probably ionic. The results suggest that FB(1) is quite stable in soils and, while tightly bound, under certain environmental conditions could be released. PMID- 12537443 TI - Comparison of a direct ELISA and an HPLC method for glyphosate determinations in water. AB - A competitive direct enzyme-linked immunosorbent assay (ELISA) and high-pressure liquid chromatographic (HPLC) methods were compared in terms of accuracy and precision for the detection and quantification of glyphosate-spiked Nanopure, tap, and river waters. The ELISA had a detection limit of 0.6 ng mL(-)(1) and a linear working range of 1-25 ng mL(-)(1), whereas the HPLC method had a detection limit of 50 ng mL(-)(1) and a linear working range of 100-10000 ng mL(-)(l). No statistically significant differences (95% confidence interval) were found between the ELISA and HPLC analysis of the three water matrixes. The coefficients of variation obtained with the ELISA in tap water were between 10 and 19%, whereas the coefficients of variation for the HPLC analysis were between 7 and 15%. The use of ELISA for the analysis of glyphosate in water is a cost-effective and reliable method capable of meeting water quality guidelines established for Europe and North America. PMID- 12537444 TI - Residue analysis of tetracyclines and their metabolites in eggs and in the environment by HPLC coupled with a microbiological assay and tandem mass spectrometry. AB - Tetracyclines are widely used in farm animals. This can cause drug residues in products of animal origin and, after excretion of these substances, in animal slurry and in soil fertilized with that slurry. In this paper, we present a method based on a microbiological assay coupled with HPLC for the detection of oxytetracycline, tetracycline, and chlortetracycline in eggs. After a simple liquid extraction of the samples and HPLC separation, fractions were collected on microtiter plates, and the tetracyclines were analyzed using the Staphylococcus aureus assay. This method was able to identify residues of tetracyclines in eggs at a level set by regulatory agencies (i.e., 200 microg/kg). In addition, it was shown that the described microbiological method can be used as a screening assay for the detection of tetracyclines and possible biologically active metabolites in animal slurry and soil samples. Employing the same extraction procedure, it was demonstrated that LC-MS-MS allowed the quantification of 20-400 microg/kg in eggs with recoveries ranging from 71 to 109% and RSDs of 3-15%. PMID- 12537445 TI - Effects of selenium supplementation on four agricultural crops. AB - Agricultural crops can be used either to remediate selenium-contaminated soils or to increase the daily selenium intake of consumers after soil supplementation using inorganic or organic selenium sources. In this study, four agricultural crops were examined for potential selenium enhancement. Soils containing tomato, strawberry, radish, and lettuce plants were supplemented with either an inorganic or an organic form of selenium. Two different soils, i.e., low Se and high Se containing, were also used. Statistically significant differences in appearance, fruit production, and fresh weights of the fruit produced were studied. Next, the amount of selenium retained in the edible fruits, nonedible plant, and soil for each was analyzed by acid digestion followed by hydride generation atomic absorption analysis. Finally, inhibition effects on the seeds of the agricultural plants were studied. The results show that supplementation with an inorganic form of selenium led to higher retention in the plants, with a maximum of 97.5% retained in the edible portion of lettuce plants. PMID- 12537446 TI - Aerobic soil metabolism of a new herbicide, LGC-42153. AB - To elucidate the fate of a new sulfonylurea herbicide, LGC-42153 [N-((4,6 dimethoxypyrimidin-2-yl)aminocarbonyl)-2-(1-methoxyacetoxy-2-fluoropropyl)-3 pyridinesulfonamide], in soil, an aerobic soil metabolism study was carried out for 120 days with [(14)C]LGC-42153 applied to a loamy soil. The material balance ranged from 90.7 to 101.5% of applied herbicide. The half-life of [(14)C]LGC 42153 was calculated to be approximately 9.0 days. The degradation products resulted from the cleavage of the sulfonylurea bridge. The metabolites identified during the study were N-((4,6-dimethoxypyrimidin-2-yl)aminocarbonyl)-2-(1-hydroxy 2-fluoropropyl)-3-pyridinesulfonamide, 2-(1-hydroxy-2-fluoropropyl)-3 pyridinesulfonamide, and 4,6-dimethoxy-2-aminopyrimidine. No significant volatile products or [(14)C]carbon dioxide was observed during the study. Nonextractable (14)C-residue reached 14.4-30.5% of applied material at 120 days after treatment, and radioactivity was distributed mostly in the humin and fulvic acid fractions. PMID- 12537447 TI - Quality assessment of strawberries (Fragaria species). AB - Several cultivars of strawberries (Fragaria sp.), grown under different conditions, were analyzed by both sensory and instrumental methods. The overall appreciation, as expressed by consumers, was mainly reflected by attributes such as sweetness and aroma. No strong correlation was obtained with odor, acidity, juiciness, or firmness. The sensory quality of strawberries can be assessed with a good level of confidence by measuring the total sugar level ( degrees Brix) and the total amount of volatile compounds. Sorting out samples using the score obtained with a hedonic test (called the "hedonic classification method") allowed the correlation between consumers' appreciation and instrumental data to be considerably strengthened. On the basis of the results obtained, a quality model was proposed. Quantitative GC-FID analyses were performed to determine the major aroma components of strawberries. Methyl butanoate, ethyl butanoate, methyl hexanoate, cis-3-hexenyl acetate, and linalool were identified as the most important compounds for the taste and aroma of strawberries. PMID- 12537448 TI - Differential effects of tomato (Lycopersicon esculentum mill) matrix on the volatility of important aroma compounds. AB - Significant tomato matrix effects on the volatility of certain fresh tomato odorants were found. The concentrations of odorants such as (E,E)-2,4-decadienal, beta-damascenone, and beta-ionone, in crushed fresh tomato fruit obtained by solid-phase microextraction (SPME), resulting from a tomato matrix calibration curve were 5.5-, 2-, and 12-fold higher, respectively, than those calculated by calibration based on buffer solutions. Static headspace analyses indicated that, in most cases, the tomato matrix significantly retains the odorants relative to the buffer solution. Thus, the concentration of odorants in the headspace of tomato is lower than expected compared to a simple matrix such as buffer. CaCl(2), although needed in crushed fruit tissue to block enzymatic activity, was found to interact specifically with 2-isobutylthiazole, reducing its content in the headspace by at least 6-fold. If a matrix effect is found, analysis of the odorant molecule contents in the headspace rather than in the food is recommended in order to better evaluate their access to the olfactory receptors. PMID- 12537449 TI - Influence of some technological parameters on the formation of dimethyl sulfide, 2-mercaptoethanol, methionol, and dimethyl sulfone in port wines. AB - Volatile sulfur compounds of 15 young port wines and 12 old port wines were determined. As there is a great difference in the pool of sulfur compounds between the two groups of wines, an experimental protocol was performed to determine which technological parameter (dissolved O(2), free SO(2) levels, pH, and time/temperature) was related with the formation/consumption of these compounds. Four sulfur compounds were selected for this purpose: dimethyl sulfide, 2-mercaptoethanol, dimethyl sulfone, and methionol. The synergistic effects of increasing temperature and O(2) at lower pH had the largest impact. Dimethyl sulfide was formed during the experimental period in the presence of O(2). Dimethyl sulfone had the same behavior. Methionol decreased significantly in the presence of O(2), but no methional was formed. 2-Mercaptoethanol, considered to be an important "off-flavor" in dry wines, also decreased during the experimental period (54 days) in the presence of O(2), and the respective disulfide was formed. These results corroborate the fact that old port wine (barrel aged) never develops "off-flavors" associated with the presence of methionol (cauliflower), 2-mercaptoethanol (rubber/burnt), or methional (cooked potato). In fact, temperature and oxygen are the major factors in the consumption of these molecules. However, some notes of "quince" and "metallic" can appear during port wine aging, and these can be associated with the presence of dimethyl sulfide. PMID- 12537450 TI - Assay of aroma active components of virgin olive oils from southern Italian regions by SPME-GC/ion trap mass spectrometry. AB - An SPME-GC/ion trap method was exploited to determine the chromatogram of volatile compounds of organic olive oils of southern Italian regions. The method is based on the assay of the terminal species of the "lipoxygenase pathway", which are present in the volatile fraction of the sampled compounds. Ethyl isobutanoate was used as internal standard in either the EI or CI ionization mode. The absolute concentration values of each analyte were evaluated through good-to-excellent calibration curves. Case studies on oils obtained from different cultivars or harvesting times are presented. The quantitative data for each compound were subjected to principal component analysis to characterize the different cultivars of this work. PMID- 12537451 TI - Age-induced diminution of free radical scavenging capacity in bee pollens and the contribution of constituent flavonoids. AB - Bee-collected pollen ("bee pollen") is promoted as a health food with a wide range of nutritional and therapeutic properties. The objective of the current study is to evaluate the contribution made through the free radical scavenging capability of bee-collected floval pollens by their flavonoid/phenolics constituents, and to determine whether this capability is affected by aging. The free radical scavenging effectiveness of a bee pollen (EC(50)) as measured by the DPPH method is shown to be determined by the nature and levels of the constituent floral pollens, which can be assayed via their phenolics profiles by HPLC. Each pure floral pollen has been found to possess a consistent EC(50) value, irrespective of its geographic origin or date of collection, and the EC(50) value is determined to a large extent (ca. 50%) by the nature and the levels of the pollen's flavonoids and phenolic acids. Non-phenolic antioxidants, possibly proteins, account for the balance of the activity. Pollen aging over 3 years is demonstrated to reduce the free radical scavenging activity by up to 50% in the most active floral pollens, which tend to contain the highest levels of flavonoids/phenolic acids. It is suggested that the freshness of a bee pollen may be determined from its free radical scavenging capacity relative to that of fresh bee pollen containing the same floral pollen mix. PMID- 12537452 TI - Comparison of components released by fermented or active dried yeasts after aging on lees in a model wine. AB - Comparison of different components released during autolysis with fermented or active dried yeast, which has never been reported previously, is related in this paper. Three autolysates were elaborated with Saccharomyces cerevisiae in a model wine (pH 3.5) at 30 or 18 degrees C. Composition of the autolysate appears to depend on both the growth medium and the physiological state of the yeast. The autolysate obtained from active dried yeast presents a higher total nitrogen concentration (a factor of 1.5-1.9 for the fermented yeast autolysate), a greater proportion of free amino acids (42 vs 16-25%), the lowest proportion of oligopeptides (25 vs 31-37%) and polypeptides (27 vs 45-34%), and an absence of nucleosides. Distribution of amino acids in peptides and proteins is relatively homogeneous in the autolysates but shows significant differences in free amino acids. Parietal macromolecules (mannoproteins and glucans) are present in greater quantities in autolysates elaborated from fermented yeast. The influence of the different fractions of these autolysates (<0.5, 0.5-1, 1-10, and >10 kDa) on bacterial growth has been investigated. PMID- 12537453 TI - Lipid oxidation in herring fillets (Clupea harengus) during ice storage measured by a commercial hybrid gas-sensor array system. AB - Volatile compounds released from herring fillets (Clupea harengus) during 15 days of storage on ice have been measured with a commercial hybrid gas-sensor array system. Using partial least-squares regression modeling, the sensor responses were correlated with data from chemical analyses (lipid oxidation products and antioxidants) and sensory analyses (odor). Eight of the 16 sensors proved significant in the correlation studies: 6 metal oxide semiconductor field effect transistor (MOSFET) sensors and 2 Taguchi type sensors. Correlation coefficients for chemical and sensory data ranged from 0.9 to 0.98 and from 0.49 to 0.92, respectively, with 0.92 referring to both "sharp/acrid" and "rancid" odors. Prediction errors ranged from 8 to 14% and from 11 to 25% for the chemical and sensory measures, respectively. That the prediction errors for oxidation product formation (5-9%) were close to the analytical errors of the chemical reference methods indicated close to "optimum" performance of the gas-sensor system. The sensor system predicted the storage time of the herring with a 1-day error. Results illustrate high potential of the gas-sensor technology in rapid nondestructive quality determination of ice-stored herring. PMID- 12537454 TI - Gelation of chicken pectoralis major myosin and heat-denatured beta lactoglobulin. AB - Thermal, rheological, and microstructural properties of myosin (1 and 2% protein) were compared to mixtures of 1% myosin and 1% heat-denatured beta-lactoglobulin aggregates (myosin/HDLG) and 1% myosin and 1% native beta-lactoglobulin (myosin/beta-LG) in 0.6 M NaCl and 0.05 M sodium phosphate buffer, pH 6.0, 6.5, and 7.0 during heating to 71 degrees C. Thermal denaturation patterns of myosin and myosin/HDLG were similar except for the appearance of an endothermic peak at 54-56 degrees C in the mixed system. At pH 7.0, 2% myosin began to gel at 48 degrees C and had a storage modulus (G') of 500 Pa upon cooling. Myosin/HDLG (2% total protein) had a gel point of 48 degrees C and a G' of 650 Pa, whereas myosin/beta-LG had a gel point of 49 degrees C but the G' was lower (180 Pa). As the pH was decreased, the gel points of myosin and myosin/HDLG decreased and the G' after cooling increased. The HDLG was incorporated within the myosin gel network, whereas beta-LG remained soluble. PMID- 12537455 TI - Influence of sucrose on droplet flocculation in hexadecane oil-in-water emulsions stabilized by beta-lactoglobulin. AB - The influence of sucrose on the flocculation stability of hydrocarbon oil-in water emulsions stabilized by a globular protein was examined using laser diffraction. Salt (150 mM NaCl) and sucrose (0-40 wt %) were added to n hexadecane oil-in-water emulsions stabilized by beta-lactoglobulin (beta-Lg, pH 7.0) either before or after isothermal heat treatment (30-95 degrees C for 20 min). When salt was added to emulsions before heat treatment, appreciable droplet flocculation was observed below the thermal denaturation temperature of the adsorbed beta-Lg (T(m) approximately 70 degrees C), and more extensive flocculation was observed above T(m). On the other hand, when salt was added to emulsions after heat treatment, appreciable droplet flocculation still occurred below T(m), but little flocculation was observed above T(m). Addition of sucrose to the emulsions increased T(m) and either promoted or suppressed droplet flocculation depending on whether it was added before or after heat treatment. These results are interpreted in terms of the influence of sucrose on protein conformational stability, protein-protein interactions, and the physiochemical properties of aqueous solutions. This study has important implications for the formulation and production of protein stabilized oil-in-water emulsions. PMID- 12537456 TI - Drying kinetics of calcium caseinate. AB - Drying is a major component of the cost of making caseinate-based films. We determined the drying curves for making calcium caseinate/glycerol films at low and high relative humidity at 21-34 degrees C. The drying curves exhibited a very long constant rate period followed by a single falling rate period. Much of the drying was in the constant rate period and preceded the actual film formation. Normally, calcium caseinate solutions are dried from about 5% solids, but it was possible to start with a more concentrated solution, 10% solids, to avoid much of the constant rate period. The resulting films were equal to those prepared starting at high initial moisture. An estimate of the drying costs indicated it is much cheaper to start with the more concentrated solutions. PMID- 12537457 TI - Purification and characterization of collagenolytic proteases from the hepatopancreas of northern shrimp (Pandalus eous). AB - Three gelatinolytic proteases (A1, A2, and B) were purified using a synthetic substrate, DNP-Pro-Gln-Gly-Ile-Ala-Gly-Gln-d-Arg, from the hepatopancreas of Northern shrimp (Pandalus eous) by several chromatographic steps involving hydroxyapatite column chromatography, gel filtration on Superdex75, and ion exchange chromatography on a MonoQ column. Collagenolytic proteases A2 and B, but not protease A1, were demonstrated to digest native porcine type I collagen at 25 degrees C and pH 7.5. Further characterizations of these two collagenolytic proteases showed that the pH optimum of enzyme A2 against DNP-peptide was found to be 11, whereas that of enzyme B was 8.5. The optimum temperature ranged between 40 and 45 degrees C for both enzymes, although enzyme B appeared to be thermally more stable than enzyme A2 at pH 7.5. Both enzymes were strongly inhibited by PMSF and antipain, which suggests that they belong to collagenolytic serine proteases. PMID- 12537458 TI - Characteristics of salt-fermented sauces from shrimp processing byproducts. AB - A salt-fermented sauce from shrimp processing byproducts (heads, shells, and tails) was prepared and characterized. Three types of sauces were prepared; sauce C, with 30 g of salt/100 g of byproduct (high salt); sauce E, with 30 g of salt and 0.2 g of sodium erythorbate (high salt); and sauce L, with 20 g of salt, 0.2 g of sodium erythorbate, 6 g of sorbitol, 0.5 mL of lactic acid, and 5 mL of ethanol (low salt). Sauces C and E showed higher exopeptidase activities than sauce L, whereas sauce L showed the highest endopeptidase activity. After 3 months of fermentation, the amino N content of sauce increased from 150-200 to 500-600 mg/100 g and the nonprotein nitrogen content increased from 300 to 950 1050 mg/100 g. Volatile basic nitrogen content increased significantly from 18 to 60 mg/100 g. The total carotenoids retained in sauces C, E, and L were 26.3, 76.2, and 73%, respectively, thus indicating that the addition of sodium erythorbate to sauces E and L retarded oxidation. Water activities of sauces C, E, and L were 0.753, 0.751, and 0.773, respectively. According to the omission test, the taste of sauces was influenced by the content of free amino acids, mainly glutamic acid and aspartic acid. All three sauces examined showed a 35% higher total amino acid content than commercial salt-fermented shrimp sauces. Therefore, shrimp processing byproducts may lend themselves to the preparation of high-quality salt-fermented sauces. PMID- 12537459 TI - Caffeic acid derivatives in the roots of yacon (Smallanthus sonchifolius). AB - Five caffeic acid derivatives were found in the roots of yacon, Smallanthus sonchifolius (Poepp. and Endl.) H. Robinson, Asteraceae, as the major water soluble phenolic compounds. The structures of these compounds were determined by analysis of spectroscopic data. Two of these were chlorogenic acid (3 caffeoylquinic acid) and 3,5-dicaffeoylquinic acid, common phenolic compounds in plants of the family Asteraceae. Three were esters of caffeic acid with the hydroxy groups of aldaric acid, derived from hexose. The structure of the aldaric moiety was determined by hydrolysis and comparison of NMR spectra with those of standard aldaric acids. The compounds were novel caffeic acid esters of altraric acid: 2,4- or 3,5-dicaffeoylaltraric acid, 2,5-dicaffeoylaltraric acid, and 2,3,5 or 2,4,5-tricaffeoylaltraric acid. PMID- 12537460 TI - Changes in sugars, acids, and volatiles during ripening of koubo [Cereus peruvianus (L.) Miller] fruits. AB - The columnar cactus Cereus peruvianus (L.) Miller, Cactaceae (koubo), is grown commercially in Israel. The unripe fruits are green, and the color changes to violet and then to red when the fruit is fully ripe. The content of soluble sugars was found to increase 5-fold during ripening. Glucose and fructose were the main sugars accumulated in the fruit pulp, and each increased from 0.5 to 5.5 g/100 g fresh weight during ripening. The polysaccharides content decreased during ripening from 1.4 to 0.4 g/100 g fresh weight. The titratable acidity decreased and the pH increased during ripening. The major organic acid found in the fruit was malic acid, which decreased from 0.75 g/100 g fresh weight at the mature green stage to 0.355 g/100 g fresh weight in ripe fruits. Citric, succinic, and oxalic acids were found in concentrations lower than 0.07 g/100 g fresh weight. Prominent accumulation of aroma volatiles occurred toward the end of the ripening process. The main volatile found in the ripe fruit was linalool, reaching concentrations of 1.5-3.5 microg/g fresh weight. PMID- 12537461 TI - Acrylamide in foods: occurrence, sources, and modeling. AB - Acrylamide in food products-chiefly in commercially available potato chips, potato fries, cereals, and bread-was determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Samples were homogenized with water/dichloromethane, centrifuged, and filtered through a 5 kDa filter. The filtrate was cleaned up on mixed mode, anion and cation exchange (Oasis MAX and MCX) and carbon (Envirocarb) cartridges. Analysis was done by isotope dilution ([D(3)]- or [(13)C(3)]acrylamide) electrospray LC-MS/MS using a 2 x 150 mm (or 2 x 100 mm) Thermo HyperCarb column eluted with 1 mM ammonium formate in 15% (or 10% for the 2 x 100 mm column) methanol. Thirty samples of foods were analyzed. Concentrations of acrylamide varied from 14 ng/g (bread) to 3700 ng/g (potato chips). Acrylamide was formed during model reactions involving heating of mixtures of amino acids and glucose in ratios similar to those found in potatoes. In model reactions between amino acids and glucose, asparagine was found to be the main precursor of acrylamide. Thus, in the reaction between nitrogen-15 (amido)-labeled asparagine and glucose, corresponding (15)N-labeled acrylamide was formed. The yield of the model reaction is approximately 0.1%. PMID- 12537462 TI - Flavone C-glycoside, phenolic acid, and nitrogen contents in leaves of barley subject to organic fertilization treatments. AB - From the leaves of barley, Hordeum vulgare, one new flavone C-glucoside and three known flavone glucosides were isolated and characterized by (1)H and (13)C NMR and MALDI-TOF-MS. The novel flavone C-glucoside was isovitexin 7-O-beta-[6' "-O (E)-p-coumaroyl]glucoside (6' "-coumaroylsaponarin), and the known compounds were isovitexin 7-O-beta-[6' "-O-(E)-feruloyl]glucoside, isoorientin 7-O-beta-[6' "-O (E)-feruloyl]glucoside, and tricin 7-O-beta-glucoside. The sum of all the flavone glycosides and soluble phenolic acids in the leaves decreased with increased rate of plant nutrients given in animal manure and with increased crop yield. All of the major phenylpropanoids showed the same general response to nutrient level. The concentration of nitrogen in the leaves was not directly related to nutrient application or to contents of phenylpropanoids. PMID- 12537463 TI - Distribution of cadmium-binding components in flax (Linum usitatissimum L.) seed. AB - The distribution of cadmium- (Cd-) binding components in flaxseed (cultivar NorMan) containing 0.526 ppm (ng/mg) Cd was investigated. Proteins extracted from dehulled, defatted flaxseed were fractionated by anion-exchange and size exclusion chromatography. The contents of Cd and other metals, UV/visible spectral characteristics, and amino acid compositions of these fractions were analyzed. Over 66% of the eluted Cd was recovered by 0.1 M NaCl elution from DEAE Sephacel, in a thiol-rich fraction representing only 7% of the extracted proteins. Sephadex G50 size-exclusion chromatography of this 0.1 M NaCl fraction concentrated most of the Cd in a low-molecular-weight peak eluting at V(t). About 72% of the extracted flaxseed proteins eluted from DEAE-Sephacel at 0.25 M NaCl and contained only 25% of the eluted Cd. Because the major Cd-binding fraction is a minor constituent of flaxseed, these results indicate the potential to isolate flaxseed's major storage protein with a low Cd content. PMID- 12537464 TI - Structural changes in plasma circulating fibrinogen after moderate beer consumption as determined by electrophoresis and spectroscopy. AB - The effects of short-term moderate beer consumption (MBC) on plasma circulating fibrinogen (PCF) in patients suffering from coronary atherosclerosis were investigated by use of 2-dimensional electrophoresis (2-DE), circular dichroism (CD), and Fourier transform infrared spectroscopy (FT-IR). Forty-eight volunteers after coronary bypass surgery were divided into experimental (EG) and control (CG) groups, each of 24. Patients of the EG group consumed 330 mL of beer/day (about 20 g of alcohol) for 30 consecutive days, and CG volunteers drank mineral water instead of beer. Blood samples were collected before and after the experiment. In 21 out of 24 patients after beer consumption the plasma circulating fibrinogen was compromised: changes in its secondary structure were found. These changes were expressed in relatively low electrophoretic mobility and charge heterogeneity, decrease in alpha-helix and increase in beta-sheet, and in slight shift of amide I and II bands. Our findings indicate that one of the positive benefits of moderate beer consumption is to diminish the production of fibrinogen and its stability, which reduces the potential risk exerted by this protein. Thus, in most of beer-consuming patients some qualitative structural changes in plasma circulating fibrinogen were detected. PMID- 12537465 TI - Stabilizing effect of ascorbic acid on flavan-3-ols and dimeric procyanidins from cocoa. AB - Cocoa flavanols and procyanidins have numerous biological activities. It is known that (-)-epicatechin, (+)-catechin, epicatechin-(4beta-8)-epicatechin (dimer B2), and epicatechin-(4beta-6)-epicatechin (dimer B5) are unstable at physiologic pH, degrading almost completely within several hours, whereas they are relatively stable at pH 5.0. The present study investigated the effects of ascorbic and citric acid on the stability of monomers and dimers in simulated intestinal juice (pH 8.5) and in sodium phosphate buffer (pH 7.4). The addition of ascorbic acid to the incubation mixture significantly increased the stability of the monomers and dimers, whereas the addition of citric acid provided no protective effects. LC-MS showed that with the degradation of dimer B2 and dimer B5, doubly linked A type dimers were formed. The present results, although not directly transferable to in vivo conditions, suggest that ascorbic acid may stabilize cocoa flavanols and procyanidins in the intestine where the pH is neutral, or alkaline, before absorption. PMID- 12537466 TI - Raisin dietary fiber composition and in vitro bile acid binding. AB - Raisins are dried grapes that are popular shelf-stable snacks. Three commercially important types of raisins were studied: sun-dried (natural), artificially dried (dipped), and sulfur dioxide-treated (golden) raisins. Dietary fiber composition was analyzed by AACC method 32-25. Polysaccharides were hydrolyzed, and the resulting sugars were analyzed by colorimetric and gas chomatographic methods. Fructans were measured with a colorimetric kit assay. Total dietary fiber values agreed with published values, with pectins and neutral polysaccharides of mannose and glucose residues predominating. Dipped raisins had over 8% fructans. No fructans were found in fresh grapes. Raisin types varied in their ability to bind bile acids in vitro. Coarsely chopped raisins bound more bile than did finely chopped or whole raisins. PMID- 12537467 TI - Concentrations of arsenic, cadmium, mercury, and lead in common foods and estimated daily intake by children, adolescents, adults, and seniors of Catalonia, Spain. AB - This study was designed to estimate the dietary intake of arsenic (As), cadmium (Cd), mercury (Hg), and lead (Pb) by the general population of Catalonia, Spain. The concentrations of these elements were determined in food samples randomly acquired in seven cities of Catalonia between June and August 2000. A total of 11 food groups were included in the study. As, Cd, Hg, and Pb levels were measured by ICP-MS and AAS. The dietary intake of these elements was determined by a total diet study. Calculations were carried out on the basis of recent data on the consumption of the selected food items. Trace element intake was estimated for five population groups: children, adolescents, male and female adults, and seniors. The highest dietary intakes of As (223.6 microg/day), Cd (15.7 microg/day), Hg (21.2 microg/day), and Pb (28.4 microg/day) corresponded to male adults. For all analyzed elements, fish and shellfish was the group showing the highest contribution to the respective intakes. In comparison with previous results, a general decrease in As, Cd, Hg, and Pb intake has occurred. The dietary intake of these elements was also compared with the provisional tolerable weekly intake (PTWI). Dietary intakes of As, Cd, Hg, and Pb by the population of Catalonia are currently well below the respective PTWIs. PMID- 12537468 TI - Solid state hydrolysis/polycondensation of alkoxysilane: access to crystal-like silicon-based hybrid materials. AB - Transformation of a crystallized molecular solid to a periodically organized covalent extended solid was obtained by a solvent-free process of an organosilane. Hydrolysis and further polycondensation occur in the solid state; in addition this route leads to crystal-like silicon-based hybrid materials with a higher periodicity than material prepared by sol-gel process. PMID- 12537469 TI - DNA polymerase-mediated DNA synthesis on a TNA template. AB - TNA, or threose nucleic acid, is capable of Watson-Crick base pairing with DNA, RNA, and TNA; coupled with its chemical simplicity, this suggests that TNA is a possible progenitor of RNA. As an initial step toward developing the molecular tools necessary to investigate the functional capabilities of TNA by in vitro selection, we have screened a variety of DNA polymerases for activity on a TNA template. We report that despite having a repeating unit that is one atom shorter than that of DNA, several polymerases showed surprisingly good ability to copy limited stretches of TNA. PMID- 12537470 TI - Experimental and computational evidence for a boron-assisted, sigma-bond metathesis pathway for alkane borylation. AB - Photoejection of one CO ligand from isolated CpM(CO)n+1BR2 (n = 1: M = Fe, Ru; n = 2: M = Mo,W; R2 = catecholate or pinacolate) compounds produces a coordinatively unsaturated 16 e- intermediate, a cyclic dioxaboryl transition metal complex, that can efficiently and selectively initiate regioselective C-H bond activation and can be used in the functionalization of alkanes. This chemistry appears distinct from that reported previously for related CpM(CO)n complexes of alkyl and aryl ligands. We show here by a combination of experimental and theoretical studies that the "unoccupied" p orbital of dioxaboryl ligands are intimately involved in the C-H bond activation step and that this hydrogen transfer to boron occurs by a boron-assisted, metal-mediated sigma-bond metathesis. The "unoccupied" p orbital of boron lowers the energy of the transition state and the intermediates by accepting electron density from the metal. The metal-bound borane then rotates, transfers back through a sigma-bond metathesis to capture the alkyl, and leaves the metal hydride. PMID- 12537471 TI - Competition between C-C and C-H insertion in prototype transition metal hydrocarbon reactions. AB - The competition between C-C and C-H insertion in model transition-metal reactions with cyclopropane and propene (C3H6) was studied as a function of total energy. Insertion of neutral transition metal atoms M (= Y, Zr, Nb, and Mo*) into the C-C bonds of cyclopropane led to formation of MCH2 + C2H4, whereas C-H insertion produced MC3H4 + H2. The measured product branching ratios verify the relative potential energy barrier heights for C-C and C-H insertion predicted by ab initio calculations. PMID- 12537472 TI - Highly unusual effects of pi-conjugation extension on the molecular linear and quadratic nonlinear optical properties of ruthenium(II) ammine complexes. AB - We have used several techniques, including hyper-Rayleigh scattering and Stark spectroscopy, to investigate the effects of polyene chain length on the optical properties of complexes containing ruthenium(II) electron donor groups and pyridinium electron acceptors. In marked contrast with all other known donor acceptor polyenes, conjugation extension beyond a single double bond in the dipolar complexes studied leads to blue-shifting of the intramolecular charge transfer absorptions. Furthermore, the static first hyperpolarizabilities beta0 become maximized with trans-1,3-butadienyl linkages and then decrease in complexes with three CH=CH bonds. Our results clearly demonstrate that the molecular engineering criteria for metal-containing nonlinear optical chromophores can differ dramatically from those for purely organic compounds. PMID- 12537473 TI - Zinc chalcogenolate complexes as capping agents in the synthesis of ternary II II'-VI nanoclusters: structure and photophysical properties of [(N,N' tmeda)5Zn5Cd11Se13(SePh)6(thf)2]. AB - We report the synthesis of the complex (N,N'-tmeda)Zn(SeSiMe3)2 (1), whose pendant trimethylsilyl moieties result in a powerful reagent for the generation of ternary MM'E materials. Reaction of 1 with (PnPr3)2Cd(OAc)2 and PhSeSiMe3 has led to the formation of the ternary ZnCdSe nanocluster [(N,N tmeda)5Zn5Cd11Se13(SePh)6(thf)2] (2). The cluster consists of a CdSe core capped by ZnSe2 units, and represents the first example of a structurally characterized II-II'-VI nanocluster. Utilization of an analogous (trimethylsilyl)tellurolate precursor leads to the formation of [(N,N-tmeda)5Zn5Cd11Te13(TePh)6(thf)] (3). The absorption spectra of 2 and 3 exhibit bands that have been assigned to "excitonic" transitions. The clusters are also luminescent at room temperature, exhibiting "band-edge" luminescence both in solution and in the solid state. PMID- 12537474 TI - UV Raman spectrum of 1,3-dimethylcyclopentenyl cation adsorbed in zeolite H-MFI. AB - The first Raman spectrum of an adsorbed carbenium ion has been measured: The 1,3 dimethylcyclopentenyl cation adsorbed in zeolite H-MFI. 1,3-Dimethylcyclopentenyl cation has been observed as a component of the hydrocarbon pool formed during the methanol-to-gasoline process catalyzed by zeolite H-MFI. The Raman shifts recorded for 1,3-dimethylcyclopentenyl cation are in remarkable agreement with computer calculations of the vibrational band positions for the isolated cation. This agreement suggests that the cation is unperturbed by interactions with the zeolite pore walls so that Raman spectra of free or solution-phase hydrocarbons can be used to identify these same species adsorbed in zeolite pores. PMID- 12537475 TI - Control of thiolate nucleophilicity and specificity in zinc metalloproteins by hydrogen bonding: lessons from model compound studies. AB - A single hydrogen bond between an amide N-H and a thiolate sulfur in model complexes designed to mimic the binding site of zinc thiolate proteins, is shown to reduce the reactivity of the thiolate toward electrophiles by up to 2 orders of magnitude. In addition a single such bond is also sufficient to achieve nearly 100% regiospecificity of reaction between a strong, and hence inherently indiscriminate, alkylating agent like trimethyl oxonium tetrafluoroborate and a single sulfur in a dithiolate construct. The importance of these results in understanding how two systems such as the zinc fingers of the GATA family and the Escherichia coli DNA repair protein Ada which share the same pseudotetrahedral structure and tetrascysteinyl ligation around the zinc can fulfill such widely divergent (structural vs reactive) roles and how specificity of reaction in multithiolate-containing systems can be achieved is discussed. PMID- 12537476 TI - An extraordinarily rapid polymerization of vinylpentafluorocyclopropane: highly stereo- and regioselective synthesis of unsaturated fluoropolymers. AB - Vinylpentafluorocyclopropane 1 was prepared from the reaction of 1,1,2-trifluoro 4-bromobutene and hexafluoropropylene oxide at 190 degrees C, following by treatment with KOH. 1 is stable at low temperature (-40 degrees C) for 7 years, but it rearranged readily to 2,3,3,4,4-pentafluorocyclopentene-1, 2, at above 80 degrees C (Ea = 28.7 kcal/mol). Under radical conditions, 1 extraordinarily rapidly polymerizes to give highly crystalline Z-fluoropolyolefin (CF2CF2CF=CHCH2)n, 3, which is very useful for cross-linking and grafting but difficult to obtain by other means. The stereochemistry of 3 was further confirmed by radical addition of iodine to 1 to form Z-ICF2CF2CF=CHCH2I, 4, exclusively. The rapid polymerization with high stereoselectivity and regioselectivity could be rationalized by effects of a favorable polar transition state of a high ring strain and electron-deficient pentafluorocyclopropyl and a relative electron-rich double bond of 1. PMID- 12537477 TI - Unusual reactivity of molecular oxygen with pi-allylnickel(N-heterocyclic carbene) chloride complexes. AB - One-pot preparation and characterization of the first reported nickel(II) complexes containing a single imidazol-2-ylidene ligand are reported. Subsequent treatment of these complexes with molecular oxygen at room temperature results in rapid formation of a mu-hydroxo dimer and alpha,beta-unsaturated carbonyl compounds. Mechanistic investigations indicate that this reaction proceeds via reversible oxygen binding followed by rate-limiting decomposition of the resulting metal-oxygen species. PMID- 12537478 TI - Carbon-carbon bonds by hydrolytic enzymes. AB - Enzymes are efficient catalysts in synthetic chemistry, and their catalytic activity with unnatural substrates in organic reaction media is an area attracting much attention. Protein engineering has opened the possibility to change the reaction specificity of enzymes and allow for new reactions to take place in their active sites. We have used this strategy on the well-studied active-site scaffold offered by the serine hydrolase Candida antarctica lipase B (CALB, EC 3.1.1.3) to achieve catalytic activity for aldol reactions. The catalytic reaction was studied in detail by means of quantum chemical calculations in model systems. The predictions from the quantum chemical calculations were then challenged by experiments. Consequently, Ser105 in CALB was targeted by site-directed mutagenesis to create enzyme variants lacking the nucleophilic feature of the active site. The experiments clearly showed an increased reaction rate when the aldol reaction was catalyzed by the mutant enzymes as compared to the wild-type lipase. We expect that the new catalytic activity, harbored in the stable protein scaffold of the lipase, will allow aldol additions of substrates, which cannot be reached by traditional aldolases. PMID- 12537480 TI - Doubly N-confused hexaphyrin: a novel aromatic expanded porphyrin that complexes bis-metals in the core. AB - Meso-hexakis(pentafluorophenyl)-substituted doubly N-confused hexaphyrins and their metal complexes were synthesized for the first time, and the structures were elucidated by X-ray single-crystal analyses. The free base form of oxidized hexaphyrin (5) had two preorganized N3O pockets in the macrocyclic core, where a hydrogen-bonding network was formed to keep the molecule planar (the mean plane deviation is 0.054 A). The formation of a planar bis-Cu(II) complex was confirmed by UV/vis, magnetic susceptibility measurements, and X-ray crystallography. The bis-Ni(II) complex (7), on the other hand, was distored from planarity, but it changed to the planar structure upon solvent (acetonitrile) coordination as judged by the observation of a sharp Soret-like band in absorption spectra and the X-ray structures of the complexes. PMID- 12537479 TI - A new turn structure for the formation of beta-hairpins in peptides. AB - Previous studies by Gellman and co-workers have elegantly shown that mirror-image beta-turns based upon d-Pro-Gly are especially good at stabilizing beta-hairpins and have demonstrated that peptide 1 [Arg-Trp-Gln-Tyr-Val-d-Pro-Gly-Lys-Phe-Thr Val-Gln-NH2] folds into a well-defined beta-hairpin [Espinosa, J. F.; Gellman, S. H. Angew. Chem., Int. Ed. 2000, 39, 2330-2333]. The present study establishes that the amino acid ornithine (Orn) also forms a turn structure that is excellent at stabilizing beta-hairpins when linked through the delta-amino group and that this turn is comparable to d-Pro-Gly in ability to induce beta-hairpin formation. Thus, 1H NMR chemical shift and NOE studies establish that Orn-containing analogue 2 [Arg-Trp-Gln-Tyr-Val-deltaOrn-Lys-Phe-Thr-Val-Gln-NH2] is comparable in structure to peptide 1. The present study also establishes that the Orn turn is superior to Asn-Gly turns and that replacement of the deltaOrn with epsilonLys or d-deltaOrn generates structures that do not fold significantly. PMID- 12537481 TI - Elucidation of the structure of a highly efficient blue electroluminescent material. AB - We determined that the structure of the previously published highly efficient blue electroluminescent borate complex, lithium tetra(2-methyl-8 hydroxyquinolato) boron, does not contain boron. Rather, it is the lithium salt of 2-methyl-8-hydroxyquinoline. PMID- 12537482 TI - A new strategy for neurochemical photodelivery: metal-ligand heterolytic cleavage. AB - A new strategy to build caged-compounds is presented. The approach is based on heterolytic photocleavage of a metal-ligand bond in a coordination compound. A ruthenium polypyridine complex, containing the neurocompound 4-amino pyridine (4AP) is used as the core of the phototrigger. The biomolecule is released by irradiation with visible light (>480 nm). The liberated 4AP promotes the activation of a leech neuron by means of blocking its K+ channels. The syntesis, characterization, and the inherent advantages of this method are discussed. PMID- 12537483 TI - Protonation and bromination of an osmabenzyne: reactions leading to the formation of new metallabenzynes. AB - The reactivities of benzynes and metal-carbyne complexes are normally associated with the triple bond units. However, we have now found that electrophiles do not attack the formal osmium-carbon triple bond of osmabenzyne complex 1. Instead, 1 undergoes electrophilic substitution reactions-the typical reactions of aromatic systems. PMID- 12537484 TI - Selective, catalytic carbon-carbon bond activation and functionalization promoted by late transition metal catalysts. AB - The selective catalytic activation and functionalization of carbon-carbon bonds in a series of substituted cyclopropane substrates has been developed using commercially available transition metal catalysts. Catalytic hydrogenation and olefination procedures, tolerant of a range of functional groups, have been discovered. Introduction of a chelate-assisting substituent such as [PPh2] is effective in altering the kinetic selectivity and lowering the activation barrier for the catalytic processes. PMID- 12537485 TI - Templating and supersaturation-driven anti-templating: principles of biomineral architecture. AB - The structural synergy between biominerals (CaCO(3), hydroxyapatite) and biosubstrates were examined for the first time. The templating effect of substrate and a newly identified supersaturation-driven interfacial structure mismatch effect were identified in the context of a new nucleation model. It follows that the heterogeneous nucleation which corresponds to a good structural match and synergy between biominerals and substrates will promote an ordered, compact, and tough complex biomineral structure, and occur only at low supersaturations, whereas at high supersaturations the heterogeneous nucleation associated with a poor structural match and synergy between biominerals and substrates will become dominant due to supersaturation-driven interfacial structural mismatch. The latter normally results in a disordered and porous structure. A phenomenon, so-called microgravity-driven homogeneous nucleation, was also examined. It turns out that microgravity will suppress convection and consequently promote homogeneous-like nucleation during biomineralization. This could be responsible for microgravity-induced osteoporousis. PMID- 12537486 TI - DNA hybridization detection with water-soluble conjugated polymers and chromophore-labeled single-stranded DNA. AB - A sensor is provided that detects single-stranded deoxyribonucleic acid (ssDNA) with a specific base sequence. The ssDNA sequence sensor comprises an aqueous solution containing a cationic water-soluble conjugated polymer [in this case, poly(9,9-bis(6'-N,N,N-trimethylammonium)-hexyl)-fluorene phenylene), 1] with a ssDNA labeled with a dye (in this case, fluorescein). The emission of light from the sensor solution with the wavelength characteristic of the probe oligonucleotide indicates the presence of ssDNA with a specific base sequence complementary to that of the probe ssDNA-fluorescein. Maximum energy transfer from 1 to the signaling chromophore occurs when the ratio of polymer chains to DNA strands is approximately 1:1. Energy transfer from 1 results in a fluorescein emission that is more intense than that observed by direct excitation of the chromophore. Furthermore, the decrease in energy transfer upon addition of electrolyte indicates that electrostatic forces dominate the interactions between 1 and DNA. PMID- 12537487 TI - Molecular dynamics and quantum chemical studies on the catalytic mechanism of Delta5-3-ketosteroid isomerase: the catalytic diad versus the cooperative hydrogen bond mechanism. AB - To further understand Delta(5)-3-ketosteroid isomerase (KSI) catalysis, we carried out molecular dynamics (MD) simulations of the KSI dimer ligated with a substrate and reaction intermediate analogue and high level ab initio calculations on relevant enzymatic reaction models. Simulation of the enzyme substrate complex dimer systems showed asymmetric dynamics between the two monomers, in which the hydrogen bond pattern between the substrate and active site residues in the first and the second subunits supported the cooperative hydrogen bond (CH) and the catalytic diad (CD) mechanisms, respectively. On the other hand, only the CH mechanism was supported in the MD simulation of the enzyme-intermediate complex dimer. From MP2/6-31+G**//RHF/6-31G** calculations, we found the kinetic barriers for the two reaction mechanisms were similar. The CH route afforded a greater stabilization to the enolate intermediate than did the CD counterpart. Thus, the present computational studies indicate that the CH mechanism would be favored over the CD one in the catalytic action of KSI. However, the latter could not be ruled out conclusively because of the explicit appearance of a CD configuration in the MD trajectories of the enzyme-substrate complex and because of the similar intrinsic activation barrier for the CH and CD mechanisms. The appearance of configurations that favor the CD pathway is rationalized in terms of a model in which the KSI-substrate complex does not have a strong preference for one hydrogen bonding pattern over another, while the KSI intermediate complex favors a cooperative hydrogen bond pattern in order to stabilize the reaction intermediate. This hypothesis is supported by the ab initio calculations which indicate that the CH intermediate is more stable than the CD one by approximately 6.3 kcal/mol. PMID- 12537488 TI - Formation of pyrene dimer radical cation in DNA reflecting DNA dynamics in the time range of 1 micros to 1 ms. AB - Doubly pyrene (Py)-conjugated oligodeoxynucleotides (ODNs) were synthesized and used for measurement of the formation rates of Py dimer radical cation (Py(2)(.+)) upon one-electron oxidation during the pulse radiolyses. Formation of Py radical cation (Py(.+)) in the time scale of less than 5 micros was monitored at 470 nm after an electron pulse during pulse radiolysis of D(2)O solution of doubly Py-conjugated ODN in the presence of K(2)S(2)O(8). Concomitant with the decay of Py(.+), formation of Py(2)(.+) with an absorption peak at 1500 nm (charge resonance band) was observed in the time range of approximately 100 micros. The formation rate of Py(2)(.+) in DNA reflected the dynamics of DNA which allows the interaction between Py(.+) and Py, since transiently formed DNA structure is trapped by the attractive charge resonance (CR) interaction to give Py(2)(.+). The formation rate of Py(2)(.+) with a characteristic CR absorption band in the near-infrared (near-IR) region was demonstrated to be useful to obtain the structural and dynamical information of transiently formed DNA in the time range of 1 micros to 1 ms. PMID- 12537489 TI - Combined use of NMR relaxation measurements and hydrodynamic calculations to study protein association. Evidence for tetramers of low molecular weight protein tyrosine phosphatase in solution. AB - We describe a novel method for determining weak association constants of oligomeric protein complexes formed transiently under equilibrium conditions. This type of equilibrium process is recognized as being biologically important, but generally hard to study. Heteronuclear spin relaxation rates measured at multiple protein concentrations are analyzed using relaxation rates predicted from hydrodynamic calculations, yielding equilibrium constants and structural characterization of the protein complexes. The method was used to study the oligomerization equilibrium of bovine low molecular weight protein tyrosine phosphatase. X-ray structures of monomeric and dimeric forms of the protein have been reported previously. Using longitudinal and transverse (15)N relaxation rates measured at four different protein concentrations, we detected the monomer, dimer, and a previously unknown tetramer and measured the dissociation constants of the equilibria involving these species. A comparison of experimental and predicted relaxation rates for individual backbone amide (15)N spins enabled delineation of the tetramerization interface. The results suggest a novel concept for substrate modulation of enzymatic activity based on a "supramolecular proenzyme". The fast and reversible switching of the "supramolecular proenzyme" would have obvious advantages for the regulation of enzymes involved in cell signaling pathways. PMID- 12537490 TI - Relative reactivity of peracids versus dioxiranes (DMDO and TFDO) in the epoxidation of alkenes. A combined experimental and theoretical analysis. AB - Comparative analysis of the calculated gas-phase activation barriers (DeltaE++) for the epoxidation of ethylene with dimethyldioxirane (DMDO) and peroxyformic acid (PFA) [15.2 and 16.4 kcal/mol at QCISD(T)// QCISD/6-31+G(d,p)] and E-2 butene [14.3 and 13.2 kcal/mol at QCISD(T)/6-31G(d)//B3LYP/6-311+G(3df,2p)] suggests similar oxygen atom donor capacities for both oxidants. Competition experiments in CH(2)Cl(2) solvent reveal that DMDO reacts with cyclohexene much faster than peracetic acid/acetic acid under scrupulously dried conditions. The rate of DMDO epoxidation is catalyzed by acetic acid with a reduction in the classical activation barrier of 8 kcal/mol. In many cases, the observed increase in the rate for DMDO epoxidation in solution may be attributed to well established solvent and hydrogen-bonding effects. This predicted epoxidative reactivity for DMDO is not consistent with what has generally been presumed for a highly strained cyclic peroxide. The strain energy (SE) of DMDO has been reassessed and its moderated value (about 11 kcal/mol) is now more consistent with its inherent gas-phase reactivity toward alkenes in the epoxidation reaction. The unusual thermodynamic stability of DMDO is largely a consequence of the combined geminal dimethyl- and dioxa-substitution effects and unusually strong C-H and C-CH(3) bonds. Methyl(trifluoromethyl)dioxirane (TFDO) exhibits much lower calculated activation barriers than DMDO in the epoxidation reaction (the average DeltaDeltaE++ values are about 7.5 kcal/mol). The rate increase relative to DMDO of approximately 10(5), while consistent with the higher strain energy for TFDO (SE approximately 19 kcal/mol) is attributed largely to the inductive effect of the CF(3) group. We have also examined the effect of alkene strain on the rate of epoxidation with PFA. The epoxidation barriers are only slightly higher for the strained alkenes cyclopropene (DeltaE++ = 14.5 kcal/mol) and cyclobutene (DeltaE++ = 13.7 kcal/mol) than for cyclopentene (DeltaE++ = 12.1 kcal/mol), reflecting the fact there is little relief of strain in the transition state. Alkenes strained by twist or pi-bond torsion do exhibit much lower activation barriers. PMID- 12537491 TI - Generation of a bacterium with a 21 amino acid genetic code. AB - We have generated a completely autonomous bacterium with a 21 amino acid genetic code. This bacterium can biosynthesize a nonstandard amino acid from basic carbon sources and incorporate this amino acid into proteins in response to the amber nonsense codon. The biosynthetic pathway for the amino acid p-aminophenylalanine (pAF) as well as a unique pAF synthetase and cognate tRNA were added to Escherichia coli. Denaturing gel electrophoresis and mass spectrometric analysis show that pAF is incorporated into myoglobin with fidelity and efficiency rivaling those of the common 20 amino acids. This and other such organisms may provide an opportunity to examine the evolutionary consequences of adding new amino acids to the genetic repertoire, as well as generate proteins with new or enhanced biological functions. PMID- 12537492 TI - Solid-phase chemical synthesis of phosphonoacetate and thiophosphonoacetate oligodeoxynucleotides. AB - Phosphonoacetate and thiophosphonoacetate oligodeoxynucleotides were prepared via a solid-phase synthesis strategy. Under Reformatsky reaction conditions, novel esterified acetic acid phosphinodiamidites were synthesized and condensed with appropriately protected 5'-O-(4, 4'-dimethoxytrityl)-2'-deoxynucleosides to yield 3'-O-phosphinoamidite reactive monomers. These synthons when activated with tetrazole were used with an automated DNA synthesizer to prepare phosphonoacetic acid modified internucleotide linkages on controlled pore glass. The phosphinoacetate coupling products were quantitatively oxidized at each step with (1S)-(+)-(10-camphorsulfonyl)oxaziridine or 3H-1,2-benzodithiol-3-one-1,1-dioxide to produce mixed sequence phosphonoacetate and thiophosphonoacetate oligodeoxynucleotides with an average per cycle coupling efficiency of greater than 97%. Completely deprotected, modified oligodeoxynucleotides were purified by reverse-phase HPLC and characterized by ion exchange HPLC, (31)P NMR, and MALDI/TOF mass spectroscopy. Both analogues were stable toward hydrolysis with snake venom phosphodiesterase and stimulated RNase H1 activity. PMID- 12537493 TI - Functionalization and kinetic stabilization of the [4]paracyclophane system and aromaticity of its extremely bent benzene ring. AB - Kinetic stabilization of the [4]paracyclophane skeleton by the introduction of substituents, which serve to sterically hinder reactions at the reactive bridgehead sites, and properties of the resultant [4]paracyclophanes are investigated in this study. Modification of the property of [4]paracyclophane by functionalization is also intended. [4]Paracyclophanes are designed to be derived from the corresponding Dewar benzene isomers via their photochemical aromatization, and the requisite 1,4-bridged Dewar benzenes bearing sterically demanding functional groups are prepared. Irradiation of these precursors under matrix isolation at 77 K leads to the formation of [4]paracyclophanes, which exhibit characteristic electronic absorption spectra. The half-lives of the generated species vary widely from less than 1 min at -90 degrees C to 0.5 h at 20 degrees C, depending on the type of substituents and the pattern of substitution. One of the derivatives, 24, is stable enough and its content in the irradiated mixture is high enough to permit the measurement of the (1)H NMR spectrum. The recorded spectrum, which is reproduced very well by theoretical calculations using the GIAO method at the hybrid HF-DFT (B3LYP/6-31+G*) level, suggests the sustenance of rather strong diatropicity in its severely bent benzene moiety. Calculations on the bent benzene whose geometry is constrained to that calculated for 24 support that aromaticity is retained to a significant extent as compared to that of planar benzene, as judged by the magnetic criteria of aromaticity, that is, diamagnetic susceptibility exaltation and nucleus independent chemical shift. The reason for the retention of aromaticity despite the severe bending of the benzene ring is discussed. Cyclophane 24 is so strained that it exceeds the corresponding Dewar benzene precursor in energy and thermally reverts to the latter with a half-life of 15 +/- 5 min at -20 degrees C (DeltaG++ = 18.3 +/- 0.3 kcal mol(-1)). PMID- 12537494 TI - Novel photoacid generators for photodirected oligonucleotide synthesis. AB - Photodirected oligonucleotide synthesis uses either direct or indirect light dependent 5'-deprotection. Both have been reported to give lower stepwise synthetic yields than conventional methods. The deficiency appears to be due to incomplete deprotection at the oligonucleotide 5'-position and, additionally in the case where photodirection is indirect and uses photogenerated photoacid to effect 5'-detritylation, the depurinating effects of strong acid. We have developed novel photosensitive-2-nitrobenzyl esters that on irradiation with near UV light generate alpha-chloro-substituted acetic acids, such as trichloroacetic acid, which are widely and successfully used in conventional solid-phase oligonucleotide synthesis. alpha-Phenyl-4,5-dimethoxy-2 nitrobenzyltrichloroacetate and alpha-phenyl-4,5-dimethoxy-2,6 dinitrobenzyltrichloroacetate showed appropriate photochemical characteristics and were used for photodirected synthesis of a variety of oligonucleotides, including (T)(5), TATAT, TGTGT, (T)(10), (AT)(5), (CT)(5) (GT)(5), and (TGCAT)(2) on a modified Millipore Expedite DNA synthesizer. The outcomes were compared with those obtained by use of directly added trichloroacetic acid (conventional synthesis). The stepwise yields for the two methods were essentially identical. PMID- 12537495 TI - Supramolecular catalysis of the enantiodifferentiating [4 + 4] photocyclodimerization of 2-anthracenecarboxylate by gamma-cyclodextrin. AB - 2-Anthracenecarboxylic acid (AC) makes a very stable 1:2 inclusion complex with gamma-cyclodextrin (gamma-CDx) (K(1) = 161 +/- 25 M(-1), K(2) = 38 500 +/- 3300 M(-1) at 25 degrees C). The formation of the 1:2 inclusion complex accelerated the photocyclodimerization of AC. The 1:2 inclusion could be clearly verified by UV-vis, CD, and (1)H NMR spectroscopies. Although these spectroscopies provide little information about the structural isomers of the inclusion complex, there should be several structural isomers of the 1:2 inclusion complex which have a different longitudinal orientation of the guest molecules in the cavity. The isomer distribution of the photodimerization product primarily depends on the population of these orientational isomers of the 1:2 inclusion complex in the ground state before photoreaction, because, in the lifetime of the excited singlet state, exchanging the orientation is impossible. The enantioselectivity of the photodimerization originates from the difference in the stability of the diastereomeric pair of orientational isomers of the 1:2 inclusion complex in the ground state, which are the precursors of the enantiomers of a specific chiral cyclodimer. The ee of a chiral cyclodimer 2 was 32% at 25 degrees C and was enhanced by lowering the temperature to 41% at 0 degrees C. This is the highest value reported for the asymmetric photodimerization in solution. PMID- 12537496 TI - Mapping the potential energy surface of the tolylcarbene rearrangement in the inner phase of a hemicarcerand. AB - Photolysis of p-tolyldiazirine (6) in the inner phase of a hemicarcerand with four butane-1,4-dioxy linker groups (5) in C(6)D(5)CD(3) at 77 K yields the 5 methylcycloheptatetraene hemicarceplex 5 circle 3b in 41% together with innermolecular reaction products resulting from an insertion of transient p tolylcarbene (1b) into an acetal C-H or linker C-O bond of 5 and from the addition of 1b to an aryl unit of 5. The yield of incarcerated 3b increased up to 67% if 6 is photolyzed inside a hemicarcerand with deuterated spanners and butane 1,4-dioxy linker groups (d(48)-5). Hemicarceplex 5 circle 3b is not formed if the photolysis is carried out in CDCl(3). Incarcerated 3b is stable at room temperature in the absence of oxygen and is characterized by 1D and 2D NMR spectroscopy. In the presence of oxygen, 3b reacts quantitatively to yield toluene and CO(2). Upon heating solutions of d(48)-5 circle 3b in C(6)D(5)CD(3), 3b rearranges to 1b and m-tolylcarbene (18). Both tolylcarbenes immediately react with the surrounding host. From a product analysis and the measured rate constants for the thermal decomposition of d(48)-5 circle 3b in the temperature range 70-102 degrees C, the activation parameters for the 3b to 1b and 3b to 18 rearrangements are calculated (3b to 1b: DeltaG(373)++ = 27.3 +/- 1.4 kcal/mol, DeltaH(373)++ = 26.4 +/- 1.0 kcal/mol, TDeltaS(373)++ = -0.9 +/- 1.0 kcal/mol; 3b to 18: DeltaG(373)++ = 27.8 +/- 1.4 kcal/mol, DeltaH(373)++ = 19.7 +/- 1.0 kcal/mol, TDeltaS(373)++ = 8.1 +/- 1.0 kcal/mol). These values are compared with those calculated by Geise and Hadad at the B3LYP/6-311+G** level of theory (Geise, C. M.; Hadad, C. M. J. Org. Chem. 2002, 67, 2532-2540). The slightly higher inner phase activation free energy of the 3b to 18 rearrangement is explained through steric constraints imposed by the surrounding hemicarcerand on the transition state. The enthalpy-entropy compensation observed for the 3b to 18 rearrangement is discussed and interpreted as a result of a hemicarcerand and solvent reorganization along the reaction coordinate. It is taken as indirect evidence for the intermediacy of 2-methylbicyclo[4.1.0]hepta-2,4,6-triene in the 3b to 18 rearrangement. PMID- 12537497 TI - Multistate/multifunctional systems. A thermodynamic, kinetic, and photochemical investigation of the 4'-dimethylaminoflavylium compound. AB - The 4'-dimethylaminoflavylium ion in aqueous solution undergoes an intricate network of chemical reactions controlled by pH and light excitation. It is shown that nine different forms are involved, including two species that are not present in previously investigated compounds of the flavylium family. The thermodynamic and kinetic constants of the equilibria and interconversion processes have been obtained by pH jump (included stopped-flow) experiments. The photochromic properties exhibited by the trans/cis chalcone forms have been investigated. The peculiar aspect of 4'-dimethylaminoflavylium, as compared to previously investigated compounds of the same family, is a close to planarity structure, as demonstrated by the X-ray analysis on the parent 4'-aminoflavylium compound (2.3 degrees torsion angle between the benzopyrylium and benzene ring). The results obtained show that the flavylium cation is strongly stabilized by the electron-donor character of the dimethylamino substituent on the benzene ring. The donor-acceptor interaction makes both the protonation of the amino group and the hydration of the flavylium cation difficult, with consequences on the tautomerization and cis/trans isomerization reactions. The multistate/multifunctional properties of 4'-dimethylaminoflavylium have been discussed in the frame of write-lock-read-unlock-erase cycles. PMID- 12537498 TI - Stereochemical memory in the regioselective and diastereoselective rearrangement of tricyclo[3.3.0.0 2,4]octanes (housanes) by electron transfer (1,3 cyclopentanediyl radical cations) and acid (cyclopentyl carbenium ions) and silver-ion catalysis. AB - The electron-transfer-catalyzed rearrangement of the housanes 5 affords regioselectively only the two cyclopentenes 6 (CH(3) migration) and 7 (R migration) by 1,2-migration of the two groups at the methano bridge to the methyl terminus. The 1,2-shift of the CH(3) group prevails, and the rearrangement ratio is essentially insensitive to the migratory aptitude of the R substituent. This stereochemical memory effect derives from the conformational impositions on the stereoelectronic requirements during the 1,2-migration in the 1,3-radical-cation intermediates. Similar regioselectivities and diastereoselectivities are observed for the TFA-catalyzed and silver(I)-ion-promoted rearrangements, whereas the rearrangement catalyzed by HClO(4) affords a complete reversal in the product selectivity and both the regioselectivity and the diastereoselectivity are much reduced. Migration to the phenyl terminus is favored to afford the 6' and 7' cyclopentenes, of which the former (CH(3) migration) dominates. For the minor regioisomer, only the cyclopentene 6 is formed by an exclusive 1,2-shift of the CH(3) group. This dichotomy in product selectivities is rationalized in terms of two distinct mechanisms for the various activation modes: a common one for the electron-transfer-induced, TFA-catalyzed, and silver(I)-ion-promoted rearrangements and a different one for HClO(4). PMID- 12537499 TI - Molecular tectonics. Use of the hydrogen bonding of boronic acids to direct supramolecular construction. AB - Tetraboronic acids 1 and 2 have four -B(OH)(2) groups oriented tetrahedrally by cores derived from tetraphenylmethane and tetraphenylsilane. Crystallization produces isostructural diamondoid networks held together by hydrogen bonding of the -B(OH)(2) groups, in accord with the tendency of simple arylboronic acids to form cyclic hydrogen-bonded dimers in the solid state. Five-fold interpenetration of the networks is observed, but 60% and 64% of the volumes of crystals of tetraboronic acids 1 and 2, respectively, remain available for the inclusion of disordered guests. Guests occupy two types of interconnected channels aligned with the a and b axes; those in crystals of tetraphenylmethane 1 measure approximately 5.9 x 5.9 A(2) and 5.2 x 8.6 A(2) in cross section at the narrowest points, whereas those in crystals of tetraphenylsilane 2 are approximately 6.4 x 6.4 A(2) and 6.4 x 9.0 A(2). These channels provide access to the interior and permit guests to be exchanged quantitatively without loss of crystallinity. Because the Si-C bonds at the core of tetraboronic acid 2 are longer (1.889(3) A) than the C-C bonds at the core of tetraboronic acid 1 (1.519(6) A), the resulting network is expanded rationally. By associating to form robust isostructural networks with predictable architectures and properties of porosity, compounds 1 and 2 underscore the usefulness of molecular tectonics as a strategy for making ordered materials. PMID- 12537500 TI - Effects of hydrogen bonding on metal ion-promoted intramolecular electron transfer and photoinduced electron transfer in a ferrocene-quinone dyad with a rigid amide spacer. AB - A ferrocene-quinone dyad (Fc-Q) with a rigid amide spacer and Fc-(Me)Q dyad, in which the amide proton acting as a hydrogen-bonding acceptor is replaced by the methyl group, are employed to examine the effects of hydrogen bonding on both the thermal and the photoinduced electron-transfer reactions. The hydrogen bonding of the semiquinone radical anion with the amide proton in Fc-Q(.-) produced by the electron-transfer reduction of Fc-Q is indicated by the significant positive shift of the one-electron reduction potential of Fc-Q. The hyperfine coupling constants of Fc-Q(.-) also indicate the existence of hydrogen bonding, agreeing with those predicted by the density functional calculation. The hydrogen-bonding dynamics in the photoinduced electron transfer from the ferrocene (Fc) to the quinone moiety (Q) in Fc-Q have been successfully detected in the femtosecond laser flash photolysis experiments. Thermal intramolecular electron transfer from Fc to Q in Fc-Q and Fc-(Me)Q also occurs efficiently in the presence of metal ions in acetonitrile at 298 K. The hydrogen bond formed between the semiquinone radical anion and the amide proton in Fc-Q results in remarkable acceleration of the rate of metal ion-promoted electron transfer as compared to the rate of Fc (Me)Q in which hydrogen bonding is prohibited. The metal ion-promoted electron transfer rates are well correlated with the binding energies of superoxide ion metal ion complexes, which are derived from the g(zz) values of the ESR spectra. PMID- 12537501 TI - Gas-phase SN2 and bromine abstraction reactions of chloride ion with bromomethane: reaction cross sections and energy disposal into products. AB - Reaction cross sections and product velocity distributions are presented for the bimolecular gas-phase nucleophilic substitution (S(N)2) reaction Cl(-) + CH(3)Br -> CH(3)Cl + Br(-) as a function of collision energy, 0.06-24 eV. The exothermic S(N)2 reaction is inefficient compared with phase space theory (PST) and ion dipole capture models. At the lowest energies, the S(N)2 reaction exhibits the largest cross sections and symmetrical forward/backward scattering of the CH(3)Cl + Br(-) products. The velocity distributions of the CH(3)Cl + Br(-) products are in agreement with an isotropic PST distribution, consistent with a complex mediated reaction and a statistical internal energy distribution of the products. Above 0.2 eV, the velocity distributions become nonisotropic and nonstatistical, exhibiting CH(3)Cl forward scattering between 0.2 and 0.6 eV. A rebound mechanism with backward scattering above 0.6 eV is accompanied by a new rising feature in the CH(3)Cl + Br(-) cross sections. The competitive endothermic reaction Cl(-) + CH(3)Br --> CH(3) + ClBr(-) rises from its thermochemical threshold at 1.9 +/- 0.4 eV, showing nearly symmetrically scattered products just above threshold and strong backward scattering above 3 eV associated with a second feature in the cross section. PMID- 12537502 TI - Theoretical investigation of the apparently irregular behavior of pt-pt nuclear spin-spin coupling constants. AB - One-bond Pt-Pt nuclear spin-spin coupling constants J(Pt-Pt) for closely related dinuclear Pt complexes can differ by an order of magnitude without any obvious correlation with Pt-Pt distances. As representative examples, the spin-spin couplings of the dinuclear Pt(I) complexes [Pt(2)(CO)(6)](2+) (1) and [Pt(2)(CO)(2)Cl(4)](2-) (2) have been computationally studied with a recently developed relativistic density functional method. The experimental values are (1)J((195)Pt-(195)Pt) = 5250 Hz for 2 but 551 Hz for 1. Many other examples are known in the literature. The experimental trends are well reproduced by the computations and can be explained based on the nature of the ligands that are coordinated to the Pt-Pt fragment. The difference for J(Pt-Pt) of an order of magnitude is caused by a sensitive interplay between the influence of different ligands on the Pt-Pt bond, and relativistic effects on metal-metal and metal ligand bonds as well as on "atomic orbital contributions" to the nuclear spin spin coupling constants. The results can be intuitively rationalized with the help of a simple qualitative molecular orbital diagram. PMID- 12537503 TI - Aurophilic interactions in cationic gold complexes with two isocyanide ligands. Polymorphic yellow and colorless forms of [(cyclohexyl isocyanide)2AuI](PF6) with distinct luminescence. AB - Crystallographic studies of yellow and colorless forms of [(C(6)H(11)NC)(2)Au(I)](PF(6)) show that they are polymorphs with differing, but close, contacts between the gold atoms which form extended chains. In the colorless polymorph the gold cations form linear chains with a short Au...Au contact (3.1822(3) A) indicative of an aurophilic attraction. The structure of the yellow polymorph is more complicated with four independent cations forming kinked, slightly helical chains with very short Au...Au contacts of 2.9803(6), 2.9790(6), 2.9651(6), and 2.9643(6) A. However, in the related compound, [(CH(3)NC)(2)Au(I)](PF(6)), each cation is surrounded by six hexafluorophosphate ions and there is no close Au...Au contact despite the fact that the isocyanide ligand has less steric bulk. The crystalline colorless and yellow polymorphs are both luminescent at 298 K, lambda(max): 424 nm (colorless) or 480 nm (yellow). Colorless solutions of the two polymorphs have identical absorption spectra and are nonluminescent at room temperature. Freezing solutions of [(C(6)H(11)NC)(2)Au(I)](PF(6)) produces intense luminescence which varies depending upon the solvent involved. Each polymorph melts to give a colorless but luminescent liquid which reverts to the yellow polymorph upon cooling. PMID- 12537504 TI - Role of copper ion in bacterial copper amine oxidase: spectroscopic and crystallographic studies of metal-substituted enzymes. AB - The role of the active site Cu(2+) of phenylethylamine oxidase from Arthrobacter globiformis (AGAO) has been studied by substitution with other divalent cations, where we were able to remove >99.5% of Cu(2+) from the active site. The enzymes reconstituted with Co(2+) and Ni(2+) (Co- and Ni-AGAO) exhibited 2.2 and 0.9% activities, respectively, of the original Cu(2+)-enzyme (Cu-AGAO), but their K(m) values for amine substrate and dioxygen were comparable. X-ray crystal structures of the Co- and Ni-AGAO were solved at 2.0-1.8 A resolution. These structures revealed changes in the metal coordination environment when compared to that of Cu-AGAO. However, the hydrogen-bonding network around the active site involving metal-coordinating and noncoordinating water molecules was preserved. Upon anaerobic mixing of the Cu-, Co-, and Ni-AGAO with amine substrate, the 480 nm absorption band characteristic of the oxidized form of the topaquinone cofactor (TPQ(ox)) disappeared rapidly (< 6 ms), yielding the aminoresorcinol form of the reduced cofactor (TPQ(amr)). In contrast to the substrate-reduced Cu-AGAO, the semiquinone radical (TPQ(sq)) was not detected in Co- and Ni-AGAO. Further, in the latter, TPQ(amr) reacted reversibly with the product aldehyde to form a species with a lambda(max) at around 350 nm that was assigned as the neutral form of the product Schiff base (TPQ(pim)). Introduction of dioxygen to the substrate reduced Co- and Ni-AGAO resulted in the formation of a TPQ-related intermediate absorbing at around 360 nm, which was assigned to the neutral iminoquinone form of the 2e(-)-oxidized cofactor (TPQ(imq)) and which decayed concomitantly with the generation of TPQ(ox). The rate of TPQ(imq) formation and its subsequent decay in Co- and Ni-AGAO was slow when compared to those of the corresponding reactions in Cu-AGAO. The low catalytic activities of the metal-substituted enzymes are due to the impaired efficiencies of the oxidative half-reaction in the catalytic cycle of amine oxidation. On the basis of these results, we propose that the native Cu(2+) ion has essential roles such as catalyzing the electron transfer between TPQ(amr) and dioxygen, in part by providing a binding site for 1e(-)- and 2e(-)-reduced dioxygen species to be efficiently protonated and released and also preventing the back reaction between the product aldehyde and TPQ(amr). PMID- 12537505 TI - Molecular quantum-dot cellular automata. AB - Molecular electronics is commonly conceived as reproducing diode or transistor action at the molecular level. The quantum-dot cellular automata (QCA) approach offers an attractive alternative in which binary information is encoded in the configuration of charge among redox-active molecular sites. The Coulomb interaction between neighboring molecules provides device-device coupling. No current flow between molecules is required. We present an ab initio analysis of a simple molecular system which acts as a molecular QCA cell. The intrinsic bistability of the charge configuration results in dipole or quadrupole fields which couple strongly to the state of neighboring molecules. We show how logic gates can be implemented. We examine the role of the relaxation of nuclear coordinates in the molecular charge reconfiguration. PMID- 12537506 TI - Cyclopentadiene annulated polycyclic aromatic hydrocarbons: investigations of electron affinities. AB - The adiabatic electron affinities of cyclopentadiene and 10 associated benzannelated derivatives have been predicted with both density functional and Hartree-Fock theory. These systems can also be regarded as benzenoid polycyclic aromatic hydrocarbons (PAHs) augmented with five-membered rings. Like the PAHs, the electron affinities of the present systems generally increase with the number of rings. To unequivocally bind an electron, cyclopentadiene must have at least two conventionally fused benzene rings. 1H-Benz[f]indene, a naphthalene-annulated cyclopentadiene, is predicted to have a zero-point energy corrected adiabatic electron affinity of 0.13 eV. Since the experimental E(A) of naphthalene is negative (-0.19 eV), the five-membered ring appendage contributes to the stability of the naphthalene-derived 1H-benz[f]indene radical anion significantly. The key to binding the electron is a contiguous sequence of fused benzenes, since fluorene, the isomer of 1H-benz[f]indene, with separated six membered rings, has an electron affinity of -0.07 eV. Each additional benzene ring in the sequence fused to cyclopentadiene increases the electron affinity by 0.15-0.65 eV: the most reliable predictions are cyclopentadiene (-0.63 eV), indene (-0.49 eV), fluorene (-0.07 eV), 1H-benz[f]indene (0.13 eV), 1,2 benzofluorene (0.25 eV), 2,3-benzofluorene (0.26 eV), 12H-dibenzo[b,h]fluorene (0.65 eV), 13H-indeno[1,2-b]anthracene (0.82 eV), and 1H-cyclopenta[b]naphthacene (1.10 eV). In contrast, if the six-membered ring-fusion is across the C(2)-C(3) cyclopentadiene single bond, only a single benzene is needed to bind an electron. The theoretical electron affinity of the resulting molecule, isoindene, is 0.49 eV, and this increases to 1.22 eV for 2H-benz[f]indene. The degree of aromaticity is responsible for this behavior. While the radical anions are stabilized by conjugation, which increases with the size of the system, the regular indenes, like PAHs in general, suffer from the loss of aromatic stabilization in forming their radical anions. While indene is 21 kcal mol(-1) more stable than isoindene, the corresponding radical anion isomers have almost the same energy. Nucleus independent chemical shift calculations show that the highly aromatic molecules lose almost all aromaticity when an extra electron is present. The radical anions of cyclopentadiene and all of its annulated derivatives have remarkably low C-H bond dissociation energies (only 18-34 kcal mol(-1) for the mono-, bi-, and tricyclics considered). Hydrogen atom loss leads to the restoration of aromaticity in the highly stabilized cyclopentadienyl anion congeners. PMID- 12537507 TI - Energetic and stereochemical effects of the protein environment on substrate: a theoretical study of methylmalonyl-CoA mutase. AB - QM/MM methods were used to study the isomerization step from (2R)-methylmalonyl CoA to succinyl-CoA. A pathway via a "fragmentation-recombination" mechanism is ruled out on energetic grounds. For the other radicalic pathway, involving an addition recombination step, geometries and vibrational contributions have been determined, and a barrier height of 11.70 kcal/mol was found. The effect of adjacent hydrogen-donating groups was found to reduce the energy barrier by 1-2 kcal/mol each and thus to provide a significant catalytic effect for this reaction. By means of molecular dynamics studies, the stereochemistry of the methylmalonyl-CoA mutase catalyzed reaction was examined. It is shown that TYR89 is essential for maintaining stereoselectivity of the abstraction of a hydrogen in the backreaction. The subsequent selective formation of one isomer of methylmalonyl-CoA is probably due to the presence of a bulky side chain. PMID- 12537508 TI - Theoretical analysis of the triplet excited state of the [Pt2(H2P2O5)4]4- ion and comparison with time-resolved X-ray and spectroscopic results. AB - A full understanding of the nature of excited states of transition metal complexes is important for understanding their chemical reactivity and role as intermediates in photochemically induced reactions. The ground and excited states of the [Pt(2)(pop)(4)](4-) ion are investigated using density functional theory (DFT). Calculations with different functionals employing quasi-relativistic Pauli and ZORA formalisms all predict a Pt-Pt bond shortening and a slight Pt-P lengthening upon excitation to the lowest triplet state, the latter in apparent contradiction to experimental EXAFS results. The PW86LYP functional with the ZORA relativistic treatment is found to produce good agreement with time-resolved crystallographic and spectroscopic results. A topological bond path between the Pt atoms is found in both the ground and the excited states, though the electron localization function (ELF) indicates weak Pt-Pt covalent bonding for the excited state only. The spin density is mainly localized on the Pt atoms, giving insight into the ability of the triplet excited state to abstract hydrogen and halogen atoms from organic substrates. PMID- 12537509 TI - Hydration of hydroxypyrrole influences binding of ImHpPyPy-beta-Dp polyamide to DNA. AB - Ligands which are able to recognize DNA sequence specifically are of fundamental interest as transcription controlling drugs. Recently a polyamide ligand was developed (ImHpPyPy-beta-Dp) which differentiates in a dimeric arrangement between all four possible base pair steps in the minor groove. This is a landmark for the design of DNA binding drugs because it was believed that such a recognition could only be possible in the major groove of DNA. Although the OH groups of the hydroxypyrrole (Hp) moieties of the ligands are responsible for this sequence discrimination, experiments showed that this OH group also reduces the absolute binding constant. We performed a free energy calculation by means of thermodynamic integration in order to find out the influence of this single hydroxyl on DNA binding. In our simulation, we found that the hydroxyl group reduces binding by about 1.3 kcal/mol, which is in excellent agreement with the experimentally determined value of 1.2 kcal/mol. In further MD simulations, the structural reasons for this reduction was estimated. The results of these simulations qualitatively agree with the X-ray structures, but in contrast, in the simulations both (ImHpPyPy-beta-Dp and ImPyPyPy-beta-Dp) ligand-DNA (d(CCAGTACTGG)(2)) complexes exhibit only slight structural differences. This is consistent with a recently published second pair of similar polyamide DNA crystal structures. Thus, we believe that the explanations resulting from the X-ray structures must be modified. We attribute the large structural differences between the two polyamide DNA complexes to a buffer molecule which binds only in the case of the ImHpPyPy-beta-Dp-DNA complex at the region of interest. We propose that the differential hydration of both ligands in the unbound state is responsible for the reduction of the binding constant. Additionally, we suggest an indirect readout of DNA, because of a lengthening of the Watson-Crick base pairs, which possibly contributes to the differentiation between T.A, A.T from G.C, C.G base pairs. PMID- 12537510 TI - Ion atmosphere relaxation control of electron transfer dynamics in a plasticized carbon dioxide redox polyether melt. AB - The sorption of CO(2) into the highly viscous, semisolid hybrid redox polyether melt, [Co(phenanthroline)(3)](MePEG-SO(3))(2), where MePEG-SO(3) is a MW 350 polyether-tailed sulfonate anion, remarkably accelerates charge transport in this molten salt material. Electrochemical measurements show that as CO(2) pressure is increased from 0 to 800 psi (54 atm) at 23 degrees C, the physical diffusion coefficient D(PHYS) of the Co(II) species, the rate constant k(EX) for Co(II/I) electron self-exchange, and the physical diffusion coefficient of the counterion D(COUNTERION) all increase, from 4.3 x 10(-10) to 6.4 x 10(-9) cm(2)/s, 4.1 x 10(6) to 1.6 x 10(7) M(-1) s(-1), and 3.3 x 10(-9) to 1.6 x 10(-8) cm(2)/s, respectively. Plots of log(k(EX)) versus log(D(PHYS)) and of log(k(EX)) versus log(D(COUNTERION)) are linear, showing that electron self-exchange rate constants are closely associated with processes that also govern D(PHYS) and D(COUNTERION). Slopes of the plots are 0.68 and 0.98, respectively, indicating a better linear correlation between k(EX) and D(COUNTERION). The evidence indicates that k(EX) can be controlled by relaxation of the counterion atmosphere about the Co complexes in the semisolid redox polyether melts. Because the counterion relaxation is in turn controlled by polyether "solvent" fluctuations, this is a new form of solvent dynamics control of electron transfer. PMID- 12537511 TI - A two-color laser photolysis method for determining reaction rates of short-lived intermediates by product analysis: application to the o-quinodimethane problem. AB - A time-delayed, two-color pulse laser photolysis technique was used for a kinetic study of short-lived transient species through product analysis, the determination of the rate constant of the cycloaddition of o-quinodimethane (1) and maleic anhydride (2) in room-temperature solutions. o-Quinodimethane (1) was generated from 1,2-bis[(phenylseleno)methyl]benzene (3) by the irradiation of a pulse of a KrF excimer laser (248 nm) in the presence of excess 2, and a successive pulse of a XeCl excimer laser (308 nm) was irradiated to the reaction mixture after varied delay times from 0 to 0.1 s for the decomposition of the remaining 1 to quench the cycloaddition reaction. The rate constant of the cycloaddition of 1 and 2 was 2.1 x 10(5) M(-1) s(-1), which was obtained by the analysis of the delay-time dependence of the product yields. PMID- 12537513 TI - Clinical pharmacokinetic profile of modafinil. AB - Modafinil is a unique wake-promoting agent for oral administration. Its pharmacological properties are distinct from those of other CNS agents, and it selectively targets neuronal pathways in the sleep/wake centres of the brain. After single or multiple oral doses, modafinil is readily absorbed, reaching maximum plasma concentrations at 2-4 hours after administration and pharmacokinetic steady state within 2-4 days. Its pharmacokinetics are dose independent between 200 and 600 mg/day. The elimination half-life is approximately 12-15 hours, which is largely reflective of the pharmacokinetics of the longer-lived l-enantiomer. Modafinil is primarily eliminated via metabolism, mainly in the liver, with subsequent excretion in the urine. Less than 10% of the dose is excreted as unchanged drug. Metabolism is largely via amide hydrolysis, with lesser contributions from cytochrome P450 (CYP)-mediated oxidative pathways. In patients who are renally or hepatically compromised, the elimination processes can be slowed, and in a similar manner (although to a lesser extent), elimination in the elderly may be reduced due to normal effects of aging. Because modafinil is administered concomitantly with other medications, the potential for metabolic drug-drug interactions has been examined both in vitro and in vivo. In vitro, modafinil was observed to produce a reversible inhibition of CYP2C19 in human liver microsomes. It also caused a small, but concentration-dependent, induction of CYP1A2, CYP2B6 and CYP3A4 activities and suppression of CYP2C9 activity in primary cultures of human hepatocytes. Clinical studies have been conducted to examine the potential for interactions with methylphenidate, dexamfetamine, warfarin, ethinylestradiol and triazolam. The only substantive interactions observed were with ethinylestradiol and triazolam, apparently through induction of CYP3A4, primarily in the gastrointestinal system. Overall, the results of the interaction studies suggest that modafinil has potential to affect the pharmacokinetics of drugs that are metabolised by certain CYP enzymes. Compounds that induce or inhibit CYP activity are unlikely to have major effects on the pharmacokinetics of modafinil. In summary, the results show that modafinil is a moderately long-lived drug that is well absorbed and extensively metabolised. PMID- 12537514 TI - Pharmacokinetics and pharmacodynamics of methotrexate in non-neoplastic diseases. AB - Low dose pulse methotrexate (LDMTX) therapy has become effective in the treatment of autoimmune and lymphoproliferative diseases. The pharmacokinetics of LDMTX is individually highly variable, resulting in a different systemic exposure to the drug and a variable therapeutic/toxic effect in patients. The improvements and exacerbations of disease activity in relation to the introductions and discontinuations of LDMTX therapy suggest the possible immunosuppresive and anti inflammatory properties of the drug. Because of a strong correlation between the drug pharmacokinetics and the therapeutic outcomes (pharmacodynamics), it seems to be possible to individualise the LDMTX therapy according to the results of pharmacokinetic/pharmacodynamic analysis. In the case of psoriasis, pharmacokinetic/pharmacodynamic analysis in our local study revealed a highly significant inverse relationship between PASI (expressed as a percent of the initial value) and a steady-state AUC(MTX) (area under the curve of methotrexate plasma concentrations; r(8) = -0.65, p < 0.001). The considerable inter individual variability and low intra-individual variability in MTX pharmacokinetics, supports a role for therapeutic monitoring and dose individualisation at the start of pharmacotherapy. The results of this study suggest that a steady-state AUC(MTX) value of 700 nmol x h/L and higher are associated with a significantly better success rate of antipsoriatic therapy than lower values. The preliminary results in our follow-up study suggest the statistically higher incidence of unwanted effects depending on maximum plasma concentration of the drug. Moreover, statistically significant correlation was found between the toxic effects and exposure to the drug regarding methotrexate plasma concentrations and intracellular storage in erythrocytes. However, the data are still in the process of being completed and are not yet published. PMID- 12537512 TI - The influence of sex on pharmacokinetics. AB - Biologic differences exist between men and women that can result in differences in responses to drugs. Both pharmacokinetic and pharmacodynamic differences between the sexes exist, with more data on pharmacokinetic differences. On average, men are larger than women. Body size differences results in larger distribution volumes and faster total clearance of most medications in men compared to women. Greater body fat in women (until older ages) may increase distribution volumes for lipophilic drugs in women. Total drug absorption does not appear to be significantly affected by sex although absorption rates may be slightly slower in women. Bioavailability after oral drug dosing, for CYP3A substrates in particular, may be somewhat higher in women compared to men. Bioavailability after transdermal drug administration does not appear to be significantly affected by gender; nor does protein binding. Renal processes of glomerular filtration, tubular secretion, and tubular reabsorption appear to be faster in men compared to women whether considered on a mg/kg basis or total body weight basis. Algorithms to estimate glomerular filtration rate incorporate sex as a factor; some also include weight. For hepatic processes, drugs metabolized by Phase I metabolism (oxidation, reduction, and hydrolysis via cytochrome P450's 1A, 2D6, 2E1), Phase II conjugative metabolism (glucuronidation, conjugation, glucuronyltransferases, methyltransferases, dehydrogenases) and by combined oxidative and conjugation processes are usually cleared faster in men compared to women (mg/kg basis). Metabolism by CYP2C9, CYP2C19, and N-acetyltransferase, appear to be similar in men and women (mg/kg). Clearance of p-glycoprotein substrates appear to be similar in men and women. In contrast, total clearance of a number of CYP3A substrates appear to be mildly or moderately faster (mg/kg) in women compared to men. The clinical significance of reported differences warrants consideration. Clearance reported on a per kg basis directly addresses organ or enzyme clearance. The difference in size between men and women means translating these results to clinical dosage rates should include an adjustment for body size. Unfortunately, this is not standard. Reports of sex differences that persist after considering weight may warrant further dosage adjustments. In addition, investigations are often performed in healthy fasting individuals yet medications are prescribed to patients with confounding influences of disease, co medications, diet, and social habits. The relative role of sex on pharmacokinetics as compared to genetics, age, disease, social habits and their potential interactions in the clinical setting is not yet fully known but should be routinely considered and further studied. PMID- 12537516 TI - A modified two-portion absorption model to describe double-peak absorption profiles of ranitidine. AB - BACKGROUND: The pharmacokinetics of oral drugs exhibiting double peaks cannot be adequately described by using conventional compartmental models. OBJECTIVE: To propose and evaluate a modified two-portion absorption model based on physiological and biopharmaceutical considerations to describe the double-peak concentration-time curve of ranitidine. MODEL DESIGN: The proposed model assumes that oral ranitidine is absorbed sequentially in two portions due to delayed gastric emptying, and thus includes a gut compartment in addition to the central and peripheral compartments. METHODS: Validation of the model was performed with respect to structural identifiability, parameter estimability and model applicability. Using initial estimates of parameters obtained from previous intravenous data, the model was used to fit oral ranitidine data from six subjects who manifested clear double-peak concentration-time profiles as well as from six subjects who showed irregular but apparent single-peak concentration time curves. RESULTS: Based on goodness-of-fit criteria, the model fitted well for both double-peak and single-peak concentration-time curves of ranitidine (for the two groups: weighted residual sum of squares, 0.044 +/- 0.027 and 0.054 +/- 0.036; correlation between observed and model predicted concentrations, 0.995 +/- 0.003 and 0.995 +/- 0.005). Simulation studies with concentrations generated with 10% normally distributed random error showed that all model fitted parameters had good accuracy and reasonable precision. The mean percentage bias ranged from -7.0 to 28.6%, and the coefficient of variance was within 30% for the majority of parameters compared with the theoretical values. CONCLUSION: The modified two portion absorption model may afford a useful approach to characterise the absorption phase and estimate pharmacokinetic parameters for drugs with two absorption peaks. PMID- 12537517 TI - Assessment of Markov-dependent stochastic models for drug administration compliance. AB - OBJECTIVE: There are few analytical results that describe patient compliance with drug administration regimens. The purpose of this paper is to develop and assess stochastic approaches for mathematical modelling of patient compliance with administration regimens. METHODS: Two stochastic models based on Markov-dependent random variables and on the Ising model were assessed for their ability to describe the variable nature of drug compliance. RESULTS: Both models use only experimentally accessible data, and their predictions were tested against published clinical compliance data obtained from electronic monitoring devices. The models satisfactorily fitted administration interval distribution data from several patients treated with diltiazem, a calcium channel antagonist, or zidovudine, an antiretroviral agent. The Ising model provides additional analytical expressions for the distribution of success runs and 'drug holidays' in administration regimens. These distribution predictions were tested with success run data for diltiazem and drug holiday data for two nonsteroidal anti inflammatory drugs, piroxicam and tenoxicam. CONCLUSIONS: Stochastic models can provide useful insights into drug compliance, and can be used to identify the administration patterns that are more likely to occur during drug self administration in populations. PMID- 12537519 TI - Alanine radicals, part 4: relative amounts of radical species in alanine dosimeters after exposure to 6-19 MeV electrons and 10 kV-15 MV photons. AB - The amino acid l-alpha-alanine can be used for high-precision dosimetry over a wide dose range, using EPR spectroscopy for monitoring radical concentrations. It is important, however, to understand the underlying composition of the observed EPR spectrum. In previous work, it was shown that the EPR signal from irradiated alanine consists of at least three different radical species, with the relative importance of each of these being almost independent of absorbed dose. However, it was not known whether the relative importance of each radical is independent of the radiation quality responsible for the EPR signal. In the present work, the relative contributions of the different radical species to the total EPR signal from alanine dosimeters irradiated with 6-19 MeV electrons and 10 kV-15 MV photons at a dose of 10 Gy were examined. By spectrum reconstruction using benchmark spectra generated from a simulation procedure, the relative amounts of the three different radical species were shown to be virtually independent of these radiation beam qualities. PMID- 12537518 TI - Molecular anatomy of the DNA damage and replication checkpoints. AB - Cell cycle checkpoints are signal transduction pathways that enforce the orderly execution of the cell division cycle and arrest the cell cycle upon the occurrence of undesirable events, such as DNA damage, replication stress, and spindle disruption. The primary function of the cell cycle checkpoint is to ensure that the integrity of chromosomal DNA is maintained. DNA lesions and disrupted replication forks are thought to be recognized by the DNA damage checkpoint and replication checkpoint, respectively. Both checkpoints initiate protein kinase-based signal transduction cascade to activate downstream effectors that elicit cell cycle arrest, DNA repair, or apoptosis that is often dependent on dose and cell type. These actions prevent the conversion of aberrant DNA structures into inheritable mutations and minimize the survival of cells with unrepairable damage. Genetic components of the damage and replication checkpoints have been identified in yeast and humans, and a working model is beginning to emerge. We summarize recent advances in the DNA damage and replication checkpoints and discuss the essential functions of the proteins involved in the checkpoint responses. PMID- 12537515 TI - Strategies and molecular probes to investigate the role of cytochrome P450 in drug metabolism: focus on in vitro studies. AB - Drug metabolism is the major determinant of drug clearance and, because of polymorphic or inducible expression of drug-metabolising cytochrome P450s (CYPs), is the factor most frequently responsible for interindividual differences in pharmacokinetics. A number of well characterised CYP substrates and inhibitors have been identified that allow precise measurements of individual CYP isoforms. Their use, alone or in combination, facilitates the phenotype characterisation of hepatocytes in vitro and in vivo. Two procedures are used for in vitro investigation of the metabolic profile of a drug: incubation with microsomes and incubation with metabolically competent cells. The major limitation of microsomes is that they express phase I activities, but only part of phase II activities, and can only be used for short incubation times. When intact cells are used, gene expression, metabolic pathways, cofactors/enzymes and plasma membrane are largely preserved, but fully differentiated cells such as primary cultured hepatocytes need to be used, since hepatoma cell lines have only very low and partial CYP expression. CYP-engineered cells or their microsomes ('supersomes') have made the identification of the CYPs involved in the metabolism of a drug candidate straightforward and easier. Inhibition of CYP is an undesirable feature for a drug candidate, and needs to be addressed by examining whether the drug candidate inhibits the metabolism of other compounds or whether other compounds inhibit the metabolism of the drug candidate. Such experiments can be conducted both with microsomes and in cells. The major limitation of microsomes is that inhibition parameters may not accurately reflect the situation in vivo, since the contribution of drug transport is not considered. The best picture of a potential drug-drug interaction can be obtained in metabolically competent hepatocytes. Screening of CYP inducers cannot be done in microsomes. It requires the use of a cellular system fully capable of transcribing and translating CYP genes, and can be monitored in vitro as an increase in enzyme mRNA or activity. Human hepatocytes in primary culture respond well to enzyme inducers during the first few days; this ability is lost thereafter. Rat hepatocytes are much less stable and soon become unresponsive to inducers. Hepatoma cell lines respond poorly to inducers, although the induction of a few isoenzymes has been reported. Primary cultured hepatocytes are still the unique in vitro model that allows global examination of the inductive potential of a drug. PMID- 12537520 TI - High-energy neutron spectroscopy with thick silicon detectors. AB - The high-energy neutron component of the space radiation environment in thick structures such as the International Space Station contributes to the total radiation dose received by an astronaut. Detector design constraints such as size and mass have limited the energy range of neutron spectrum measurements in orbit to about 12 MeV in Space Shuttle studies. We present a new method for high-energy neutron spectroscopy using small silicon detectors that can extend these measurements to more than 500 MeV. The methodology is based on measurement of the detector response function for high-energy neutrons and inversion of this response function with measured deposition data to deduce neutron energy spectra. We also present the results of an initial shielding study performed with the thick silicon detector system for high-energy neutrons incident on polyethylene. PMID- 12537521 TI - Lung cancer after treatment for Hodgkin's disease: focus on radiation effects. AB - Aspects of radiation-induced lung cancer were evaluated in an international study of Hodgkin's disease. The study population consisted of 227 patients with lung cancer and 455 matched controls. Unique features included dose determinations to the specific location in the lung where each cancer developed and quantitative data on both chemotherapy and tobacco use obtained from medical records. The estimated excess relative risk (ERR) per Gy was 0.15 (95% CI: 0.06-0.39), and there was little evidence of departure from linearity even though lung doses for the majority of Hodgkin's disease patients treated with radiotherapy exceeded 30 Gy. The interaction of radiation and chemotherapy that included alkylating agents was almost exactly additive, and a multiplicative relationship could be rejected (P = 0.017). Conversely, the interaction of radiation and smoking was consistent with a multiplicative relationship, but not with an additive relationship (P < 0.001). The ERR/Gy for males was about four times that for females, although the difference was not statistically significant. There was little evidence of modification of the ERR/Gy by time since exposure (after a 5-year minimum latent period), age at exposure, or attained age. Because of the very high radiation doses received by Hodgkin's disease patients and the immunodeficiency inherent to this lymphoma and that associated with chemotherapy, generalizing these findings to other populations receiving considerably lower doses of radiation should be done cautiously. PMID- 12537522 TI - Proteinuria and vascular changes after renal irradiation: the role of reactive oxygen species (ROS) and vascular endothelial growth factor (Vegf). AB - Proteinuria occurs in all degrees of radiation nephropathy and can be present without other symptoms. In this study, radiation-induced proteinuria in C3H mice demonstrated a clear dose-response relationship and was apparent before the onset of significant structural vascular changes and decreases in renal function. This suggests that proteinuria is not a secondary event due to loss of the vascular structure. In an attempt to ameliorate radiation-induced proteinuria and progressive renal failure, two factors were studied. The influence of reactive oxygen species (ROS), which are generated by infiltrating neutrophils and mediate proteinuria in models of acute glomerular injury, was the first to be investigated. Short-term administration of the reactive oxygen scavengers superoxide dismutase (SOD) and catalase did not reverse an established radiation induced proteinuria. Continuous administration of the antioxidant N acetylcysteine (NAC) also failed to inhibit this proteinuria. However, since no direct assessment of the impact of these interventions on renal redox status was made, the putative role of ROS in radiation-induced proteinuria and nephropathy remains undefined. The second factor studied was vascular endothelial growth factor (Vegf), which is suggested to be involved in glomerular vessel permeability and the development of proteinuria in some models of renal disease. Northern blot analysis of mRNA from whole kidneys did not demonstrate any increased expression of Vegf after irradiation. There was also no change in the ratio of the different Vegf isoforms (PCR analysis), either in the whole kidney or in isolated glomeruli. No significant role for Vegf was identified for radiation-induced vascular changes or proteinuria, although post-transcriptional changes cannot be excluded. PMID- 12537523 TI - The nonuniformity of antibody distribution in the kidney and its influence on dosimetry. AB - The therapeutic efficacy of radiolabeled antibody fragments can be limited by nephrotoxicity, particularly when the kidney is the major route of extraction from the circulation. Conventional dose estimates in kidney assume uniform dose deposition, but we have shown increased antibody localization in the cortex after glomerular filtration. The purpose of this study was to measure the radioactivity in cortex relative to medulla for a range of antibodies and to assess the validity of the assumption of uniformity of dose deposition in the whole kidney and in the cortex for these antibodies with a range of radionuclides. Storage phosphor plate technology (radioluminography) was used to acquire images of the distributions of a range of antibodies of various sizes, labeled with 125I, in kidney sections. This allowed the calculation of the antibody concentration in the cortex relative to the medulla. Beta-particle point dose kernels were then used to generate the dose-rate distributions from 14C, 131I, 186Re, 32P and 90Y. The correlation between the actual dose-rate distribution and the corresponding distribution calculated assuming uniform antibody distribution throughout the kidney was used to test the validity of estimating dose by assuming uniformity in the kidney and in the cortex. There was a strong inverse relationship between the ratio of the radioactivity in the cortex relative to that in the medulla and the antibody size. The nonuniformity of dose deposition was greatest with the smallest antibody fragments but became more uniform as the range of the emissions from the radionuclide increased. Furthermore, there was a strong correlation between the actual dose-rate distribution and the distribution when assuming a uniform source in the kidney for intact antibodies along with medium- to long range radionuclides, but there was no correlation for small antibody fragments with any radioisotope or for short-range radionuclides with any antibody. However, when the cortex was separated from the whole kidney, the correlation between the actual dose-rate distribution and the assumed dose-rate distribution, if the source was uniform, increased significantly. During radioimmunotherapy, the extent of nonuniformity of dose deposition in the kidney depends on the properties of the antibody and radionuclide. For dosimetry estimates, the cortex should be taken as a separate source region when the radiopharmaceutical is small enough to be filtered by the glomerulus. PMID- 12537524 TI - Oxygen depletion during and after mTHPC-mediated photodynamic therapy in RIF1 and H-MESO1 tumors. AB - During photodynamic therapy (PDT), low oxygenation levels, induced both by oxygen consumption and by vascular occlusion, can lead to an inefficient photochemical reaction that may compromise the efficacy of PDT. In the present studies, tumor oxygenation was measured before, during and after meta-tetrahydroxyphenylchlorin (mTHPC)-mediated PDT of murine RIF1 tumors and human mesothelioma xenografts (H MESO1). Tumor pO2 was measured in real time with Eppendorf polarography, and the extent of relative hypoxia at specific times was measured by immunohistochemical staining. Significant decreases in median pO2 values, as well as an increase in the number of values below 2.5 mmHg, were seen during and after PDT in RIF1 tumors, although there was a large intertumoral variation. Tumor pO2 values did not change significantly in H-MESO1 tumors. Staining with antibodies against the hypoxia marker EF3 showed significant increases in relative hypoxia after PDT in both tumor types compared with separate groups of untreated controls. Our results are consistent with PDT-induced oxygen depletion (reduced pO2) leading to an increase in relative hypoxia in RIF1 tumors. Extensive necrosis in the H-MESO1 tumors may have prevented the detection of PDT-induced hypoxia using the Eppendorf polarographic needle, whereas immunohistochemistry did reveal increases in relative hypoxia. PMID- 12537525 TI - Evaluation of the relative biological effectiveness of a clinical epithermal neutron beam using dog brain. AB - This investigation was designed to determine the relative biological effectiveness (RBE) of an epithermal neutron beam (FiR 1 beam) using the brains of dogs. The FiR 1 beam was developed for the treatment of patients with glioma using boron neutron capture therapy. Comparisons were made between the effects of whole-brain irradiation with epithermal neutrons and 6 MV photons. For irradiations with epithermal neutrons, three dose groups were used, 9.4 +/- 0.1, 10.2 +/- 0.1 and 11.5 +/- 0.2 Gy. These physical doses were given as a single exposure and are quoted at the 90% isodose. Four groups of five dogs were irradiated with single doses of 10, 12, 14 or 16 Gy of 6 MV photons to the 100% isodose. Different reference isodoses were used to obtain the most comparable dose distribution in the brain for the two different irradiation modalities. Sequential magnetic resonance images (MRI) were taken for 77-115 weeks after irradiation to detect changes in the brain. Dose-effect relationships were established for changes in the brain as detected either by MRI or by subsequent gross morphology and histology. The doses that caused a specified response in 50% of the animals (ED(50)) were calculated from these dose-effect curves for each end point, and these values were used to calculate the RBE values for the different end points. The RBE values for the FiR 1 beam, based on changes observed on MRI, were in the range 1.2-1.3. For microscopic and gross pathological lesions, the values were in the range 1.2-1.4. The corresponding RBE values for the MRI and pathological end points for the high-LET components (protons from nitrogen capture and recoil protons from fast neutrons) were in the ranges 3.5-4.0 and 3.4-4.4, respectively. This assumed a dose-rate reduction factor of 0.6 for the low-dose-rate gamma-ray component of this beam. Finally, a comparison was made between experimentally derived photon doses, for a specified end point, with calculated photon equivalent doses, which were obtained using the weighting factors for clinical studies on the epithermal neutron beam on the Brookhaven Medical Research Reactor (BNL) in New York. This indicated that the radiation-induced lesions seen in the present study were, on average, detected at a 12% lower photon dose than predicted by the use of the BNL clinical weighting factors. This indicates the need for caution in the extrapolation of results from one reactor-based epithermal neutron beam to another. PMID- 12537526 TI - Photoprotection by antioxidants against UVB-radiation-induced damage in pig skin organ culture. AB - Topically applied antioxidants constitute an important group of protective agents against skin damage induced by ultraviolet radiation. The current study was performed to investigate whether a recently developed ex vivo pig skin model was suitable for short-term studies of the mechanism(s) of UVB-radiation-induced skin damage; the protective effect of topical application of alpha-tocopherol, l ascorbic acid, alpha-lipoic acid, glutathione ethylester and N-acetylcysteine was tested. Increasing doses of the antioxidants were applied topically on ex vivo pig skin explants and allowed to penetrate for 60 min. Epidermal antioxidant bioavailability was measured before and 60 min after exposure to an ultraviolet B (UVB) radiation of 7.5 kJ/m2. Cell viability (trypan blue dye exclusion) and apoptosis were measured 48 h later in isolated keratinocytes. UVB-radiation induced epidermal lipid peroxidation was determined immediately after exposure of the skin to a UVB dose of 28 kJ/m2. All antioxidants tested became bioavailable in pig skin epidermis, and none of them were depleted after UVB-radiation exposure. Increasing doses of the antioxidants tested decreased UVB-radiation induced cell death and apoptosis. The highest doses of antioxidants prevented UVB radiation-induced lipid peroxidation; alpha-lipoic acid only tended to decrease lipid peroxidation. In conclusion, a single topical dose of the above antioxidants on ex vivo pig skin can reduce UVB-radiation-induced oxidative stress and lipid peroxidation and thereby reduce apoptotic stimuli and cell death. Furthermore, the ex vivo pig skin model was a useful tool for testing compounds for their antioxidant activity. PMID- 12537527 TI - Redox reactions obtained by gamma irradiation of quercetin methanol solution are similar to in vivo metabolism. AB - The flavonol quercetin is one of the most well-known antioxidant flavonoids. Its antioxidant potential has been studied extensively during the last 10 years, but little is known about the metabolites formed in vivo that lead to the formation of depside and small molecules such as benzoic acids. In this study, gamma irradiation of a quercetin methanol solution was used as a model of certain oxidative reactions that occur in vivo. Qercetin at concentrations ranging from 5 x 10(-5) M to 5 x 10(-3) M, was irradiated with gamma rays at doses of 2-14 kGy. Quercetin degradation was evaluated by HPLC analysis. The major radiolytic metabolite was identified as a depside by NMR and LC-MS. Formation of 3,4 dihydroxybenzoic acid was also observed. The presence of CH3O. formed during methanol radiolysis is invoked to explain depside formation. Transformation of the 8-methoxy substituted depside (Q1) to the 8-hydroxyl substituted depside (Q2) is discussed. The antioxidant properties of quercetin metabolites are evaluated according to their capacity to decrease the EPR DPPH signal and to inhibit superoxide radical formed by the enzymatic reaction (xanthine + xanthine oxidase). For both assays, the IC50 of Q2 is twice as high as that of quercetin. PMID- 12537528 TI - Second analysis of mortality of nuclear industry workers in Japan, 1986-1997. AB - IA cohort study of nuclear industry workers was initiated in 1990 to determine the possible health effects of low-level radiation. A total of 5,527 deaths were ascertained among 176,000 male workers who had been retrospectively and/or prospectively followed for an average of 7.9 years during the observation period 1986-1997. Statistical analyses were made mainly on the prospective follow-up outcome of 120,000 workers followed for an average of 4.5 years. The standardized mortality ratio (and its 95% confidence interval) was 0.94 (0.90, 0.97) for 2,934 cases of all causes combined and 0.86 (0.82, 0.91) for 1,305 cases of non malignant diseases combined, which suggested a healthy worker effect. For 1,191 cases of all cancers combined, it was 0.98 (0.93, 1.04), indicating no difference in mortality from that of the general population. In tests for trend of death rate with increasing radiation dose, no significant correlation was found for all cancers combined. For site-specific cancers, most cancers including leukemia showed no positive correlation with dose, except for cancers of the esophagus, stomach and rectum and multiple myeloma. External causes showed a significant correlation with dose. A separate questionnaire study indicated that these positive findings could be ascribed in part to lifestyle characteristics of the workers. For leukemia only, we attempted to estimate the excess relative risk per unit dose of radiation, which, with reservations because of its wide confidence interval, was within the range of variation of the risks reported in other radiation epidemiological studies. This population must be studied for a longer time and with a consideration of the possible effects of confounding factors. PMID- 12537529 TI - Evaluation of age and gender dependences of the rate of strontium elimination 25 45 years after intake: analysis of data from residents living along the Techa river. AB - The Mayak Production Association released large amounts of 90Sr into the Techa River with peak amounts in 1950-1951. Residents near the Techa River ingested an average of approximately 3,000 kBq of 90Sr. The affected people have been followed by scientists at the Urals Research Center for Radiation Medicine. The whole-body content of 90Sr of approximately 15,000 individuals has been measured over a period of 24 years (1974-1997) using a special whole-body counter. This report evaluates the gender and age dependences of individual rates of strontium elimination. Data on persons who had been measured 12 or more times were selected for study. There were 108 men and 81 women older than 30 years who met this criterion. Individual measurement results were fitted to an exponential function and grouped mean averages of the rate of strontium elimination as a function of age for each sex were derived. For men, a significant increase (from 2.8% year( 1) to 3.2% year(-1)) in the rate of strontium elimination after age 55 years is seen. For women, the increase in the rate of elimination was significant at age 45 and reached 5.8% year(-1) after the age of 60. The results may be used to develop a gender- and age-dependent model of strontium metabolism. PMID- 12537530 TI - Induction of chromatid breaks by carbon K-shell ultrasoft X rays. AB - Chromatid breaks have previously been shown to be induced in G2-phase cells after exposure to ionizing radiation (X and gamma rays) as a linear function of dose, consistent with a single-event mechanism. DNA double-strand breaks (DSBs) are thought to be the initiating lesion, and experiments with a genetically engineered cell line containing a single DSB site also indicate that a single DSB is sufficient to induce a chromatid break. Although the precise mechanism of conversion of an isolated DSB into a chromatid break is not yet understood, it is known that a proportion of chromatid breaks result from rearrangements between sister chromatids. Here we report further evidence for the single-event hypothesis for the formation of chromatid breaks. The evidence derives from experiments in which chromatid breaks have been induced by exposure of Chinese hamster cells to ultrasoft carbon K-shell X rays. Since the energy of carbon K shell X rays is not sufficient for the secondary electrons to span more than one DNA double helix, we conclude that single traversals, and hence single (complex) DSBs, are responsible for the formation of chromatid breaks. We find that, as for 60Co gamma rays, around 10% of the carbon K-shell X-ray-induced chromatid breaks have associated color switches at breakpoints, indicating that they arise through sister chromatid rearrangements. PMID- 12537531 TI - Repair of radiation-induced DNA double-strand breaks is dependent upon radiation quality and the structural complexity of double-strand breaks. AB - Mammalian cells primarily repair DSBs by nonhomologous end joining (NHEJ). To assess the ability of human cells to mediate end joining of complex DSBs such as those produced by chemicals, oxidative events, or high- and low-LET radiation, we employed an in vitro double-strand break repair assay using plasmid DNA linearized by these various agents. We found that human HeLa cell extracts support end joining of complex DSBs and form multimeric plasmid products from substrates produced by the radiomimetic drug bleomycin, 60Co gamma rays, and the effects of 125I decay in DNA. End joining was found to be dependent on the type of DSB-damaging agent, and it decreased as the cytotoxicity of the DSB-inducing agent increased. In addition to the inhibitory effects of DSB end-group structures on repair, NHEJ was found to be strongly inhibited by lesions proximal to DSB ends. The initial repair rate for complex non-ligatable bleomycin-induced DSBs was sixfold less than that of similarly configured (blunt-ended) but less complex (ligatable) restriction enzyme-induced DSBs. Repair of DSBs produced by gamma rays was 15-fold less efficient than repair of restriction enzyme-induced DSBs. Repair of the DSBs produced by 125I was near the lower limit of detection in our assay and was at least twofold lower than that of gamma-ray-induced DSBs. In addition, DSB ends produced by 125I were shown to be blocked by 3'-nucleotide fragments: the removal of these by E. coli endonuclease IV permitted ligation. PMID- 12537532 TI - Involvement of the nonhomologous end joining DNA repair pathway in the bystander effect for chromosomal aberrations. AB - Cells of mouse knockout cell lines for Ku80 (now known as Xrcc5), Ku70 (now known as G22p1), DNA-PKcs (now known as Prkdc) and PARP (now known as Adprt) were synchronized in G1 phase and exposed to very low fluences of alpha particles. The frequency of gross chromosomal aberrations was scored at the first postirradiation metaphase. At the two lowest doses examined, aberrations were induced in 4-9% of wild-type cells and 36-55% of Xrcc5-/- cells, whereas only 2 3% of the nuclei were traversed by an alpha particle and thus received any radiation exposure. G22p1-/- cells responded similarly to Xrcc5-/- cells, whereas Prkdc-/- and Adprt-/- cells showed an intermediate effect. The frequency of aberrations per nuclear traversal increased approximately 30-fold for Xrcc5-/- and G22p1-/- cells at the lowest mean dose examined (0.17 cGy), compared with 10 fold in Prkdc-/- cells and 3-fold in wild-type cells. Based on these and other findings, we hypothesize that the marked sensitization of repair-deficient bystander cells to the induction of chromosomal aberrations is a consequence of unrejoined DNA double-strand breaks occurring as a result of clustered damage arising from opposed oxidative lesions and single-strand breaks. PMID- 12537533 TI - Flow cytometry analysis of gamma-radiation-induced Epstein-Barr virus reactivation in lymphocytes. AB - Epstein-Barr virus (EBV), a member of the gamma-herpesvirus family, is involved in the development of several diseases, and the infection is believed to persist for life in latent form. Ionizing radiation at clinically relevant doses may increase the amount of virus reactivation in B cells, and the combination of radiation with stress could amplify EBV reactivation. In vitro experiments were performed on several cell lines, including EBV-positive Burkitt lymphoma cells. The presence of the immediate-early protein ZEBRA, which is a hallmark of EBV reactivation, was evaluated using flow cytometry, which enabled us to measure the percentage of ZEBRA-positive cells. The process was studied previously in the EBV positive Burkitt lymphoma cell line B95-8. Forty-eight hours after irradiation alone, 13.6 and 19.9% ZEBRA-positive cells were observed at 2 and 4 Gy, respectively, compared to the basal level of 1.85%. Thus irradiation induces EBV reactivation. The addition of a glucocorticoid (the final effector of the stress response) had no effect on EBV reactivation in our model. However, the combination of radiation and treatment with a glucocorticoid (dexamethasone) increased the expression of ZEBRA in B95-8 cells (15.8 and 28.75% of the cells was positive at 24 and 48 h after gamma irradiation, respectively). Thus the combination of gamma radiation and a glucocorticoid may play an important role in EBV reactivation. PMID- 12537534 TI - Comments on "Long-term exposure of Emicro-Pim1 transgenic mice to 898.4 MHz microwaves does not increase lymphoma incidence" by Utteridge et al., Radiat. Res. 158, 357-364 (2002). PMID- 12537535 TI - Comments on "Long-term exposure of Emicro-Pim1 transgenic mice to 898.4 MHz microwaves does not increase lymphoma incidence" by Utteridge et al., Radiat. Res. 158, 357-364 (2002). PMID- 12537536 TI - Comments on the recent publication on microwave effects on Emicro-Pim1 transgenic mice (Utteridge et al., Radiat. Res. 158, 357-364, 2002). PMID- 12537541 TI - Everything I need to know about genomics, I learned from Yogi Berra. AB - As 2002 draws to a close, I find myself contemplating the future of biology with hope, but also with uncertainty and some apprehension. When I find myself in such a spiritual quandary, I usually turn to a guru. PMID- 12537539 TI - Mouse system-N amino acid transporter, mNAT3, expressed in hepatocytes and regulated by insulin-activated and phosphoinositide 3-kinase-dependent signalling. AB - Amino acid transporters are essential for normal cell function and physiology. In the present study, we report the identification and functional and regulatory characterization of a mouse system-N amino acid transporter, mNAT3. Expression of mNAT3 in Xenopus oocytes revealed that the strongest transport activities were preferred for L-alanine. In addition, mNAT3 is an Na(+)- and pH-dependent low affinity transporter and it partially tolerates substitution of Na(+) by Li(+). mNAT3 has been found to be expressed predominantly in the liver, where it is localized to the plasma membrane of hepatocytes, with the strongest expression in those cells adjacent to the central vein, decreasing gradually towards the portal tract. Treatment of mouse hepatocyte-like H2.35 cells with insulin led to a significant increase in the expression of mNAT3, and this stimulation was associated closely with an increase in the uptake of L-alanine. Interestingly, this insulin-induced stimulatory effect on mNAT3 expression was attenuated by the phosphoinositide 3-kinase inhibitor LY294002, but not by the mitogen-activated protein kinase inhibitor PD98059, although both kinases were fully activated by insulin. The results suggest that insulin-mediated regulation of mNAT3 is likely to be mediated through a phosphoinositide 3-kinase-dependent signalling pathway. The unique expression pattern and insulin-mediated regulatory properties of mNAT3 suggest that this transporter may play an important role in liver physiology. PMID- 12537542 TI - A hat trick--Plasmodium, Anopheles and Homo. AB - The genomes of the malaria parasite, its vector and its host are now sequenced. This has been a tremendous scientific achievement. But will it offer hope to the millions who die from malaria each year? Yes, but only if combined with political will and social change. PMID- 12537543 TI - The integrated world of functional genomics. PMID- 12537544 TI - Gene function in the mammalian genome, courtesy of the mouse. PMID- 12537545 TI - The genome and the germ cell. PMID- 12537546 TI - Computational comparison of two mouse draft genomes and the human golden path. AB - BACKGROUND: The availability of both mouse and human draft genomes has marked the beginning of a new era of comparative mammalian genomics. The two available mouse genome assemblies, from the public mouse genome sequencing consortium and Celera Genomics, were obtained using different clone libraries and different assembly methods. RESULTS: We present here a critical comparison of the two latest mouse genome assemblies. The utility of the combined genomes is further demonstrated by comparing them with the human 'golden path' and through a subsequent analysis of a resulting conserved sequence element (CSE) database, which allows us to identify over 6,000 potential novel genes and to derive independent estimates of the number of human protein-coding genes. CONCLUSION: The Celera and public mouse assemblies differ in about 10% of the mouse genome. Each assembly has advantages over the other: Celera has higher accuracy in base-pairs and overall higher coverage of the genome; the public assembly, however, has higher sequence quality in some newly finished bacterial artificial chromosome clone (BAC) regions and the data are freely accessible. Perhaps most important, by combining both assemblies, we can get a better annotation of the human genome; in particular, we can obtain the most complete set of CSEs, one third of which are related to known genes and some others are related to other functional genomic regions. More than half the CSEs are of unknown function. From the CSEs, we estimate the total number of human protein-coding genes to be about 40,000. This searchable publicly available online CSEdb will expedite new discoveries through comparative genomics. PMID- 12537547 TI - A gene-expression program reflecting the innate immune response of cultured intestinal epithelial cells to infection by Listeria monocytogenes. AB - BACKGROUND: Listeria monocytogenes is a Gram-positive, facultative, intracellular bacterial pathogen found in soil, which occasionally causes serious food-borne disease in humans. The outcome of an infection is dependent on the state of the infected individual's immune system, neutrophils being key players in clearing the microorganism from the body. The first line of host defense, however, is the intestinal epithelium. RESULTS: We have examined the transcriptional response of cultured human intestinal epithelial cells to infection by L. monocytogenes, which replicates in the host cell cytoplasm and spreads from cell to cell using a form of actin-based motility. We found that the predominant host response to infection was mediated by NFkappaB. To determine whether any host responses were due to recognition of specific virulence factors during infection, we also examined the transcriptional response to two bacterial mutants; actA which is defective in actin-based motility, and prfA, which is defective in the expression of all L. monocytogenes virulence genes. Remarkably, we found no detectable difference in the host transcriptional response to the wild-type and mutant bacteria. CONCLUSIONS: These results suggest that cultured intestinal epithelial cells are capable of mounting and recruiting a powerful innate immune response to L. monocytogenes infection. Our results imply that L. monocytogenes is not specifically detected in the host cytoplasm of Caco-2 cells by intracellular signals. This suggests that entry of bacteria is mediated in the host cell post translationally, and that these bacteria seek the cytosol not only for the nutrient-rich environment, but also for protection from detection by the immune system. PMID- 12537548 TI - Hap4p overexpression in glucose-grown Saccharomyces cerevisiae induces cells to enter a novel metabolic state. AB - BACKGROUND: Metabolic and regulatory gene networks generally tend to be stable. However, we have recently shown that overexpression of the transcriptional activator Hap4p in yeast causes cells to move to a state characterized by increased respiratory activity. To understand why overexpression of HAP4 is able to override the signals that normally result in glucose repression of mitochondrial function, we analyzed in detail the changes that occur in these cells. RESULTS: Whole-genome expression profiling and fingerprinting of the regulatory activity network show that HAP4 overexpression provokes changes that also occur during the diauxic shift. Overexpression of HAP4, however, primarily acts on mitochondrial function and biogenesis. In fact, a number of nuclear genes encoding mitochondrial proteins are induced to a greater extent than in cells that have passed through a normal diauxic shift: in addition to genes required for mitochondrial energy conservation they include genes encoding mitochondrial ribosomal proteins. CONCLUSIONS: We show that overproduction of a single nuclear transcription factor enables cells to move to a novel state that displays features typical of, but clearly not identical to, other derepressed states. PMID- 12537551 TI - MRD: a microsatellite repeats database for prokaryotic and eukaryotic genomes. AB - MRD is a database system to access the microsatellite repeats information of genomes such as archea, eubacteria, and other eukaryotic genomes whose sequence information is available in public domains. MRD stores information about simple tandemly repeated k-mer sequences where k= 1 to 6, i.e. monomer to hexamer. The web interface allows the users to search for the repeat of their interest and to know about the association of the repeat with genes and genomic regions in the specific organism. The data contains the abundance and distribution of microsatellites in the coding and non-coding regions of the genome. The exact location of repeats with respect to genomic regions of interest (such as UTR, exon, intron or intergenic regions) whichever is applicable to organism is highlighted. MRD is available on the World Wide Web at http://www.ccmb.res.in/mrd and/or http://www.ingenovis.com/mrd. The database is designed as an open-ended system to accommodate the microsatellite repeats information of other genomes whose complete sequences will be available in future through public domain. PMID- 12537552 TI - Osprey: a network visualization system. AB - We have developed a software platform called Osprey for visualization and manipulation of complex interaction networks. Osprey builds data-rich graphical representations that are color-coded for gene function and experimental interaction data. Mouse-over functions allow rapid elaboration and organization of network diagrams in a spoke model format. User-defined large-scale data sets can be readily combined with Osprey for comparison of different methods. PMID- 12537549 TI - Microarray analysis of orthologous genes: conservation of the translational machinery across species at the sequence and expression level. AB - BACKGROUND: Genome projects have provided a vast amount of sequence information. Sequence comparison between species helps to establish functional catalogues within organisms and to study how they are maintained and modified across phylogenetic groups during evolution. Microarray studies allow us to determine groups of genes with similar temporal regulation and perhaps also common regulatory upstream regions for binding of transcription factors. The integration of sequence and expression data is expected to refine our current annotations and provide some insight into the evolution of gene regulation across organisms. RESULTS: We have investigated how well the protein subcellular localization and functional categories established from clustering of orthologous genes agree with gene-expression data in Saccharomyces cerevisiae. An increase in the resolution of biologically meaningful classes is observed upon the combination of experiments under different conditions. The functional categories deduced by sequence comparison approaches are, in general, preserved at the level of expression and can sometimes interact into larger co-regulated networks, such as the protein translation process. Differences and similarities in the expression between cytoplasmic-mitochondrial and interspecies translation machineries complement evolutionary information from sequence similarity. CONCLUSIONS: Combination of several microarray experiments is a powerful tool for the identification of upstream regulatory motifs of yeast genes involved in protein synthesis. Comparison of these yeast co-regulated genes against the archaeal and bacterial operons indicates that the components of the protein translation process are conserved across organisms at the expression level with minor specific adaptations. PMID- 12537553 TI - The GRID: The General Repository for Interaction Datasets. AB - We have developed a relational database, called the General Repository for Interaction Datasets (The GRID; http://biodata.mshri.on.ca/grid) to archive and display physical, genetic and functional interactions. The GRID displays data rich interaction tables for any protein of interest, combines literature-derived and high throughput interaction datasets, and is readily accessible via the World Wide Web. Interactions parsed in the GRID can be viewed in graphical form with a versatile visualization tool called Osprey. PMID- 12537554 TI - Whose genome is next? PMID- 12537555 TI - The repertoire of protein kinases encoded in the draft version of the human genome: atypical variations and uncommon domain combinations. AB - BACKGROUND: Phosphorylation by protein kinases is central to cellular signal transduction. Abnormal functioning of kinases has been implicated in developmental disorders and malignancies. Their activity is regulated by second messengers and by the binding of associated domains, which are also influential in translocating the catalytic component to their substrate sites, in mediating interaction with other proteins and carrying out their biological roles. RESULT: Using sensitive profile-search methods and manual analysis, the human genome has been surveyed for protein kinases. A set of 448 sequences, which show significant similarity to protein kinases and contain the critical residues essential for kinase function, have been selected for an analysis of domain combinations after classifying the kinase domains into subfamilies. The unusual domain combinations in particular kinases suggest their involvement in ubiquitination pathways and alternative modes of regulation for mitogen-activated protein kinase kinases (MAPKKs) and cyclin-dependent kinase (CDK)-like kinases. Previously unexplored kinases have been implicated in osteoblast differentiation and embryonic development on the basis of homology with kinases of known functions from other organisms. Kinases potentially unique to vertebrates are involved in highly evolved processes such as apoptosis, protein translation and tyrosine kinase signaling. In addition to coevolution with the kinase domain, duplication and recruitment of non-catalytic domains is apparent in signaling domains such as the PH, DAG-PE, SH2 and SH3 domains. CONCLUSIONS: Expansion of the functional repertoire and possible existence of alternative modes of regulation of certain kinases is suggested by their uncommon domain combinations. Experimental verification of the predicted implications of these kinases could enhance our understanding of their biological roles. PMID- 12537556 TI - Strong-association-rule mining for large-scale gene-expression data analysis: a case study on human SAGE data. AB - BACKGROUND: The association-rules discovery (ARD) technique has yet to be applied to gene-expression data analysis. Even in the absence of previous biological knowledge, it should identify sets of genes whose expression is correlated. The first association-rule miners appeared six years ago and proved efficient at dealing with sparse and weakly correlated data. A huge international research effort has led to new algorithms for tackling difficult contexts and these are particularly suited to analysis of large gene-expression matrices. To validate the ARD technique we have applied it to freely available human serial analysis of gene expression (SAGE) data. RESULTS: The approach described here enables us to designate sets of strong association rules. We normalized the SAGE data before applying our association rule miner. Depending on the discretization algorithm used, different properties of the data were highlighted. Both common and specific interpretations could be made from the extracted rules. In each and every case the extracted collections of rules indicated that a very strong co-regulation of mRNA encoding ribosomal proteins occurs in the dataset. Several rules associating proteins involved in signal transduction were obtained and analyzed, some pointing to yet-unexplored directions. Furthermore, by examining a subset of these rules, we were able both to reassign a wrongly labeled tag, and to propose a function for an expressed sequence tag encoding a protein of unknown function. CONCLUSIONS: We show that ARD is a promising technique that turns out to be complementary to existing gene-expression clustering techniques. PMID- 12537557 TI - Membrane-bound progesterone receptors contain a cytochrome b5-like ligand-binding domain. AB - BACKGROUND: Membrane-associated progesterone receptors (MAPRs) are thought to mediate a number of rapid cellular effects not involving changes in gene expression. They do not show sequence similarity to any of the classical steroid receptors. We were interested in identifying distant homologs of MAPR better to understand their biological roles. RESULTS: We have identified MAPRs as distant homologs of cytochrome b5. We have also found regions homologous to cytochrome b5 in the mammalian HERC2 ubiquitin transferase proteins and a number of fungal chitin synthases. CONCLUSIONS: In view of these findings, we propose that the heme-binding cytochrome b5 domain served as a template for the evolution of membrane-associated binding pockets for non-heme ligands. PMID- 12537558 TI - Supervised clustering of genes. AB - BACKGROUND: We focus on microarray data where experiments monitor gene expression in different tissues and where each experiment is equipped with an additional response variable such as a cancer type. Although the number of measured genes is in the thousands, it is assumed that only a few marker components of gene subsets determine the type of a tissue. Here we present a new method for finding such groups of genes by directly incorporating the response variables into the grouping process, yielding a supervised clustering algorithm for genes. RESULTS: An empirical study on eight publicly available microarray datasets shows that our algorithm identifies gene clusters with excellent predictive potential, often superior to classification with state-of-the-art methods based on single genes. Permutation tests and bootstrapping provide evidence that the output is reasonably stable and more than a noise artifact. CONCLUSIONS: In contrast to other methods such as hierarchical clustering, our algorithm identifies several gene clusters whose expression levels clearly distinguish the different tissue types. The identification of such gene clusters is potentially useful for medical diagnostics and may at the same time reveal insights into functional genomics. PMID- 12537559 TI - Whole-genome analysis of animal A- and B-type cyclins. AB - BACKGROUND: Multiple A- and B-type cyclins have been identified in animals, but their study is complicated by varying degrees of functional redundancy. A non essential phenotype may reflect redundancy with a known or as yet unknown gene. Complete sequencing of several animal genomes has allowed us to determine the size of the mitotic cyclin gene family and therefore to start to address this issue. RESULTS: We analyzed the Caenorhabditis elegans, Drosophila melanogaster and Homo sapiens genomes to identify known and novel A- and B-type cyclin genes and distinguish them from related pseudogenes. We find only a single functional A type cyclin gene in invertebrates but two in vertebrates. In addition to the single functional cyclin A gene, the C. elegans genome contains numerous cyclin A pseudogenes. In contrast, the number and relationship of B-type cyclins varies considerably between organisms but all contain at least one cyclin B1-like gene and a cyclin B3 gene. CONCLUSIONS: There are three conserved families of mitotic cyclins in animals: A-, B3- and B-type. The precise number of genes within the A- and B-type families varies in different organisms, possibly as an adaptation to their distinct developmental strategies. PMID- 12537560 TI - Bayesian analysis of gene expression levels: statistical quantification of relative mRNA level across multiple strains or treatments. AB - BACKGROUND: Methods of microarray analysis that suit experimentalists using the technology are vital. Many methodologies discard the quantitative results inherent in cDNA microarray comparisons or cannot be flexibly applied to multifactorial experimental design. Here we present a flexible, quantitative Bayesian framework. This framework can be used to analyze normalized microarray data acquired by any replicated experimental design in which any number of treatments, genotypes, or developmental states are studied using a continuous chain of comparisons. RESULTS: We apply this method to Saccharomyces cerevisiae microarray datasets on the transcriptional response to ethanol shock, to SNF2 and SWI1 deletion in rich and minimal media, and to wild-type and zap1 expression in media with high, medium, and low levels of zinc. The method is highly robust to missing data, and yields estimates of the magnitude of expression differences and experimental error variances on a per-gene basis. It reveals genes of interest that are differentially expressed at below the twofold level, genes with high 'fold-change' that are not statistically significantly different, and genes differentially regulated in quantitatively unanticipated ways. CONCLUSIONS: Anyone with replicated normalized cDNA microarray ratio datasets can use the freely available MacOS and Windows software, which yields increased biological insight by taking advantage of replication to discern important changes in expression level both above and below a twofold threshold. Not only does the method have utility at the moment, but also, within the Bayesian framework, there will be considerable opportunity for future development. PMID- 12537561 TI - Identification and utilization of arbitrary correlations in models of recombination signal sequences. AB - BACKGROUND: A significant challenge in bioinformatics is to develop methods for detecting and modeling patterns in variable DNA sequence sites, such as protein binding sites in regulatory DNA. Current approaches sometimes perform poorly when positions in the site do not independently affect protein binding. We developed a statistical technique for modeling the correlation structure in variable DNA sequence sites. The method places no restrictions on the number of correlated positions or on their spatial relationship within the site. No prior empirical evidence for the correlation structure is necessary. RESULTS: We applied our method to the recombination signal sequences (RSS) that direct assembly of B-cell and T-cell antigen-receptor genes via V(D)J recombination. The technique is based on model selection by cross-validation and produces models that allow computation of an information score for any signal-length sequence. We also modeled RSS using order zero and order one Markov chains. The scores from all models are highly correlated with measured recombination efficiencies, but the models arising from our technique are better than the Markov models at discriminating RSS from non RSS. CONCLUSIONS: Our model-development procedure produces models that estimate well the recombinogenic potential of RSS and are better at RSS recognition than the order zero and order one Markov models. Our models are, therefore, valuable for studying the regulation of both physiologic and aberrant V(D)J recombination. The approach could be equally powerful for the study of promoter and enhancer elements, splice sites, and other DNA regulatory sites that are highly variable at the level of individual nucleotide positions. PMID- 12537562 TI - A strategy for oligonucleotide microarray probe reduction. AB - BACKGROUND: One of the factors limiting the number of genes that can be analyzed on high-density oligonucleotide arrays is that each transcript is probed by multiple oligonucleotide probes. To reduce the number of probes required for each gene, a systematic approach to choosing the most representative probes is needed. A method is presented for reducing the number of probes per gene while maximizing the fidelity to the original array design. RESULTS: The methodology has been tested on a dataset comprising 317 Affymetrix HuGeneFL GeneChips. The performance of the original and reduced probe sets was compared in four cancer-classification problems. The results of these comparisons show that reduction of the probe set by 95% does not dramatically affect performance, and thus illustrate the feasibility of substantially reducing probe numbers without significantly compromising sensitivity and specificity of detection. CONCLUSIONS: The strategy described here is potentially useful for designing small, limited-probe genome wide arrays for screening applications. PMID- 12537563 TI - Heart-specific genes revealed by expressed sequence tag (EST) sampling. AB - BACKGROUND: Cardiovascular diseases are the primary cause of death worldwide; the identification of genes specifically expressed in the heart is thus of major biomedical interest. We carried out a comprehensive analysis of gene-expression profiles using expressed sequence tags (ESTs) to identify genes overexpressed in the human adult heart. The initial set of genes expressed in the heart was constructed by clustering and assembling ESTs from heart cDNA libraries. Expression profiles were then generated for each gene by counting their cognate ESTs in all libraries. Differential expression was assessed by applying a previously published statistical procedure to these profiles. RESULTS: We identified 35 cardiac-specific genes overexpressed in the heart, some of which displayed significant coexpression. Some genes had no previously recognized cardiac function. Of the 35 genes, 32 were mapped back onto the human genome sequence. According to Online Mendelian Inheritance in Man (OMIM), five genes were previously known as heart-disease genes and one gene was located in the locus of a bleeding disorder. Analysis of the promoter regions of this collection of genes provides the first list of putative regulatory elements associated with differential cardiac expression. CONCLUSION: This study shows that ESTs are still a powerful tool to identify differentially expressed genes. We present a list of genes specifically expressed in the human heart, one of which is a candidate for a bleeding disorder. In addition, we provide the first set of putative regulatory elements, the combination of which appears correlated with heart-specific gene expression. PMID- 12537564 TI - Identification of frequent cytogenetic aberrations in hepatocellular carcinoma using gene-expression microarray data. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is a leading cause of death worldwide. Frequent cytogenetic abnormalities that occur in HCC suggest that tumor-modifying genes (oncogenes or tumor suppressors) may be driving selection for amplification or deletion of these particular genetic regions. In many cases, however, the gene(s) that drive the selection are unknown. Although techniques such as comparative genomic hybridization (CGH) have traditionally been used to identify cytogenetic aberrations, it might also be possible to identify them indirectly from gene-expression studies. A technique we have called comparative genomic microarray analysis (CGMA) predicts regions of cytogenetic change by searching for regional gene-expression biases. CGMA was applied to HCC gene-expression profiles to identify regions of frequent cytogenetic change and to identify genes whose expression is misregulated within these regions. RESULTS: Using CGMA, 104 HCC gene-expression microarray profiles were analyzed. CGMA identified 13 regions of frequent cytogenetic change in the HCC samples. Ten of these regions have been detected in previous CGH studies (+lq, -4q, +6p, -8p, +8q, -13q, -16q, -17p, +17q, +20q). CGMA identified three additional regions that have not been previously identified by CGH (+5q, +12q, +19p). Genes located in regions of frequent cytogenetic change were examined for changed expression in the HCC samples. CONCLUSIONS: Our results suggest that CGMA predictions using gene expression microarray datasets are a practical alternative to CGH profiling. In addition, CGMA might be useful for identifying candidate genes within cytogenetically abnormal regions. PMID- 12537566 TI - JEvTrace: refinement and variations of the evolutionary trace in JAVA. AB - BACKGROUND: Details of functional speciation within gene families can be difficult to identify using standard multiple sequence alignment (MSA) methods. The evolutionary trace (ET) was developed as a visualization tool to combine MSA, phylogenetic and structural data for identification of functional sites in proteins. The method has been successful in extracting evolutionary details of functional surfaces in a number of biological systems and modifications of the method are useful in creating hypotheses about the function of previously unannotated genes. We wish to facilitate the graphical interpretation of disparate data types through the creation of flexible software implementations. RESULTS: We have implemented the ET method in a JAVA graphical interface, JEvTrace. Users can analyze and visualize ET input and output with respect to protein phylogeny, sequence and structure. Function discovery with JEvTrace is demonstrated on two proteins with recently determined crystal structures: YlxR from Streptococcus pneumoniae with a predicted RNA-binding function, and a Haemophilus influenzae protein of unknown function, YbaK. To facilitate analysis and storage of results we propose a MSA coloring data structure. The sequence coloring format readily captures evolutionary, biological, functional and structural features of MSAs. CONCLUSIONS: Protein families and phylogeny represent complex data with statistical outliers and special cases. The JEvTrace implementation of the ET method allows detailed mining and graphical visualization of evolutionary sequence relationships. PMID- 12537565 TI - Prediction and overview of the RpoN-regulon in closely related species of the Rhizobiales. AB - BACKGROUND: In the rhizobia, a group of symbiotic Gram-negative soil bacteria, RpoN (sigma54, sigmaN, NtrA) is best known as the sigma factor enabling transcription of the nitrogen fixation genes. Recent reports, however, demonstrate the involvement of RpoN in other symbiotic functions, although no large-scale effort has yet been undertaken to unravel the RpoN-regulon in rhizobia. We screened two complete rhizobial genomes (Mesorhizobium loti, Sinorhizobium meliloti) and four symbiotic regions (Rhizobium etli, Rhizobium sp. NGR234, Bradyrhizobium japonicum, M. loti) for the presence of the highly conserved RpoN-binding sites. A comparison was also made with two closely related non-symbiotic members of the Rhizobiales (Agrobacterium tumefaciens, Brucella melitensis). RESULTS: A highly specific weight-matrix-based screening method was applied to predict members of the RpoN-regulon, which were stored in a highly annotated and manually curated dataset. Possible enhancer-binding proteins (EBPs) controlling the expression of RpoN-dependent genes were predicted with a profile hidden Markov model. CONCLUSIONS: The methodology used to predict RpoN-binding sites proved highly effective as nearly all known RpoN-controlled genes were identified. In addition, many new RpoN-dependent functions were found. The dependency of several of these diverse functions on RpoN seems species-specific. Around 30% of the identified genes are hypothetical. Rhizobia appear to have recruited RpoN for symbiotic processes, whereas the role of RpoN in A. tumefaciens and B. melitensis remains largely to be elucidated. All species screened possess at least one uncharacterized EBP as well as the usual ones. Lastly, RpoN could significantly broaden its working range by direct interfering with the binding of regulatory proteins to the promoter DNA. PMID- 12537567 TI - In silico discovery of gene-coding variants in murine quantitative trait loci using strain-specific genome sequence databases. AB - BACKGROUND: The identification of genes underlying complex traits has been aided by quantitative trait locus (QTL) mapping approaches, which in turn have benefited from advances in mammalian genome research. Most recently, whole-genome draft sequences and assemblies have been generated for mouse strains that have been used for a large fraction of QTL mapping studies. Here we show how such strain-specific mouse genome sequence databases can be used as part of a high throughput pipeline for the in silico discovery of gene-coding variations within murine QTLs. As a test of this approach we focused on two QTLs on mouse chromosomes 1 and 13 that are involved in physical dependence on alcohol. RESULTS: Interstrain alignment of sequences derived from the relevant mouse strain genome sequence databases for 199 QTL-localized genes spanning 210,020 base-pairs of coding sequence identified 21 genes with different coding sequences for the progenitor strains. Several of these genes, including four that exhibit strong phenotypic links to chronic alcohol withdrawal, are promising candidates to underlie these QTLs. CONCLUSIONS: This approach has wide general utility, and should be applicable to any of the several hundred mouse QTLs, encompassing over 60 different complex traits, that have been identified using strains for which relatively complete genome sequences are available. PMID- 12537568 TI - Finishing a whole-genome shotgun: release 3 of the Drosophila melanogaster euchromatic genome sequence. AB - BACKGROUND: The Drosophila melanogaster genome was the first metazoan genome to have been sequenced by the whole-genome shotgun (WGS) method. Two issues relating to this achievement were widely debated in the genomics community: how correct is the sequence with respect to base-pair (bp) accuracy and frequency of assembly errors? And, how difficult is it to bring a WGS sequence to the accepted standard for finished sequence? We are now in a position to answer these questions. RESULTS: Our finishing process was designed to close gaps, improve sequence quality and validate the assembly. Sequence traces derived from the WGS and draft sequencing of individual bacterial artificial chromosomes (BACs) were assembled into BAC-sized segments. These segments were brought to high quality, and then joined to constitute the sequence of each chromosome arm. Overall assembly was verified by comparison to a physical map of fingerprinted BAC clones. In the current version of the 116.9 Mb euchromatic genome, called Release 3, the six euchromatic chromosome arms are represented by 13 scaffolds with a total of 37 sequence gaps. We compared Release 3 to Release 2; in autosomal regions of unique sequence, the error rate of Release 2 was one in 20,000 bp. CONCLUSIONS: The WGS strategy can efficiently produce a high-quality sequence of a metazoan genome while generating the reagents required for sequence finishing. However, the initial method of repeat assembly was flawed. The sequence we report here, Release 3, is a reliable resource for molecular genetic experimentation and computational analysis. PMID- 12537570 TI - An integrated computational pipeline and database to support whole-genome sequence annotation. AB - We describe here our experience in annotating the Drosophila melanogaster genome sequence, in the course of which we developed several new open-source software tools and a database schema to support large-scale genome annotation. We have developed these into an integrated and reusable software system for whole-genome annotation. The key contributions to overall annotation quality are the marshalling of high-quality sequences for alignments and the design of a system with an adaptable and expandable flexible architecture. PMID- 12537571 TI - Apollo: a sequence annotation editor. AB - The well-established inaccuracy of purely computational methods for annotating genome sequences necessitates an interactive tool to allow biological experts to refine these approximations by viewing and independently evaluating the data supporting each annotation. Apollo was developed to meet this need, enabling curators to inspect genome annotations closely and edit them. FlyBase biologists successfully used Apollo to annotate the Drosophila melanogaster genome and it is increasingly being used as a starting point for the development of customized annotation editing tools for other genome projects. PMID- 12537569 TI - A Drosophila full-length cDNA resource. AB - BACKGROUND: A collection of sequenced full-length cDNAs is an important resource both for functional genomics studies and for the determination of the intron-exon structure of genes. Providing this resource to the Drosophila melanogaster research community has been a long-term goal of the Berkeley Drosophila Genome Project. We have previously described the Drosophila Gene Collection (DGC), a set of putative full-length cDNAs that was produced by generating and analyzing over 250,000 expressed sequence tags (ESTs) derived from a variety of tissues and developmental stages. RESULTS: We have generated high-quality full-insert sequence for 8,921 clones in the DGC. We compared the sequence of these clones to the annotated Release 3 genomic sequence, and identified more than 5,300 cDNAs that contain a complete and accurate protein-coding sequence. This corresponds to at least one splice form for 40% of the predicted D. melanogaster genes. We also identified potential new cases of RNA editing. CONCLUSIONS: We show that comparison of cDNA sequences to a high-quality annotated genomic sequence is an effective approach to identifying and eliminating defective clones from a cDNA collection and ensure its utility for experimentation. Clones were eliminated either because they carry single nucleotide discrepancies, which most probably result from reverse transcriptase errors, or because they are truncated and contain only part of the protein-coding sequence. PMID- 12537572 TI - Annotation of the Drosophila melanogaster euchromatic genome: a systematic review. AB - BACKGROUND: The recent completion of the Drosophila melanogaster genomic sequence to high quality and the availability of a greatly expanded set of Drosophila cDNA sequences, aligning to 78% of the predicted euchromatic genes, afforded FlyBase the opportunity to significantly improve genomic annotations. We made the annotation process more rigorous by inspecting each gene visually, utilizing a comprehensive set of curation rules, requiring traceable evidence for each gene model, and comparing each predicted peptide to SWISS-PROT and TrEMBL sequences. RESULTS: Although the number of predicted protein-coding genes in Drosophila remains essentially unchanged, the revised annotation significantly improves gene models, resulting in structural changes to 85% of the transcripts and 45% of the predicted proteins. We annotated transposable elements and non-protein-coding RNAs as new features, and extended the annotation of untranslated (UTR) sequences and alternative transcripts to include more than 70% and 20% of genes, respectively. Finally, cDNA sequence provided evidence for dicistronic transcripts, neighboring genes with overlapping UTRs on the same DNA sequence strand, alternatively spliced genes that encode distinct, non-overlapping peptides, and numerous nested genes. CONCLUSIONS: Identification of so many unusual gene models not only suggests that some mechanisms for gene regulation are more prevalent than previously believed, but also underscores the complex challenges of eukaryotic gene prediction. At present, experimental data and human curation remain essential to generate high-quality genome annotations. PMID- 12537573 TI - The transposable elements of the Drosophila melanogaster euchromatin: a genomics perspective. AB - BACKGROUND: Transposable elements are found in the genomes of nearly all eukaryotes. The recent completion of the Release 3 euchromatic genomic sequence of Drosophila melanogaster by the Berkeley Drosophila Genome Project has provided precise sequence for the repetitive elements in the Drosophila euchromatin. We have used this genomic sequence to describe the euchromatic transposable elements in the sequenced strain of this species. RESULTS: We identified 85 known and eight novel families of transposable element varying in copy number from one to 146. A total of 1,572 full and partial transposable elements were identified, comprising 3.86% of the sequence. More than two-thirds of the transposable elements are partial. The density of transposable elements increases an average of 4.7 times in the centromere-proximal regions of each of the major chromosome arms. We found that transposable elements are preferentially found outside genes; only 436 of 1,572 transposable elements are contained within the 61.4 Mb of sequence that is annotated as being transcribed. A large proportion of transposable elements is found nested within other elements of the same or different classes. Lastly, an analysis of structural variation from different families reveals distinct patterns of deletion for elements belonging to different classes. CONCLUSIONS: This analysis represents an initial characterization of the transposable elements in the Release 3 euchromatic genomic sequence of D. melanogaster for which comparison to the transposable elements of other organisms can begin to be made. These data have been made available on the Berkeley Drosophila Genome Project website for future analyses. PMID- 12537574 TI - Heterochromatic sequences in a Drosophila whole-genome shotgun assembly. AB - BACKGROUND: Most eukaryotic genomes include a substantial repeat-rich fraction termed heterochromatin, which is concentrated in centric and telomeric regions. The repetitive nature of heterochromatic sequence makes it difficult to assemble and analyze. To better understand the heterochromatic component of the Drosophila melanogaster genome, we characterized and annotated portions of a whole-genome shotgun sequence assembly. RESULTS: WGS3, an improved whole-genome shotgun assembly, includes 20.7 Mb of draft-quality sequence not represented in the Release 3 sequence spanning the euchromatin. We annotated this sequence using the methods employed in the re-annotation of the Release 3 euchromatic sequence. This analysis predicted 297 protein-coding genes and six non-protein-coding genes, including known heterochromatic genes, and regions of similarity to known transposable elements. Bacterial artificial chromosome (BAC)-based fluorescence in situ hybridization analysis was used to correlate the genomic sequence with the cytogenetic map in order to refine the genomic definition of the centric heterochromatin; on the basis of our cytological definition, the annotated Release 3 euchromatic sequence extends into the centric heterochromatin on each chromosome arm. CONCLUSIONS: Whole-genome shotgun assembly produced a reliable draft-quality sequence of a significant part of the Drosophila heterochromatin. Annotation of this sequence defined the intron-exon structures of 30 known protein-coding genes and 267 protein-coding gene models. The cytogenetic mapping suggests that an additional 150 predicted genes are located in heterochromatin at the base of the Release 3 euchromatic sequence. Our analysis suggests strategies for improving the sequence and annotation of the heterochromatic portions of the Drosophila and other complex genomes. PMID- 12537575 TI - Assessing the impact of comparative genomic sequence data on the functional annotation of the Drosophila genome. AB - BACKGROUND: It is widely accepted that comparative sequence data can aid the functional annotation of genome sequences; however, the most informative species and features of genome evolution for comparison remain to be determined. RESULTS: We analyzed conservation in eight genomic regions (apterous, even-skipped, fushi tarazu, twist, and Rhodopsins 1, 2, 3 and 4) from four Drosophila species (D. erecta, D. pseudoobscura, D. willistoni, and D. littoralis) covering more than 500 kb of the D. melanogaster genome. All D. melanogaster genes (and 78-82% of coding exons) identified in divergent species such as D. pseudoobscura show evidence of functional constraint. Addition of a third species can reveal functional constraint in otherwise non-significant pairwise exon comparisons. Microsynteny is largely conserved, with rearrangement breakpoints, novel transposable element insertions, and gene transpositions occurring in similar numbers. Rates of amino-acid substitution are higher in uncharacterized genes relative to genes that have previously been studied. Conserved non-coding sequences (CNCSs) tend to be spatially clustered with conserved spacing between CNCSs, and clusters of CNCSs can be used to predict enhancer sequences. CONCLUSIONS: Our results provide the basis for choosing species whose genome sequences would be most useful in aiding the functional annotation of coding and cis-regulatory sequences in Drosophila. Furthermore, this work shows how decoding the spatial organization of conserved sequences, such as the clustering of CNCSs, can complement efforts to annotate eukaryotic genomes on the basis of sequence conservation alone. PMID- 12537576 TI - Computational analysis of core promoters in the Drosophila genome. AB - BACKGROUND: The core promoter, a region of about 100 base-pairs flanking the transcription start site (TSS), serves as the recognition site for the basal transcription apparatus. Drosophila TSSs have generally been mapped by individual experiments; the low number of accurately mapped TSSs has limited analysis of promoter sequence motifs and the training of computational prediction tools. RESULTS: We identified TSS candidates for about 2,000 Drosophila genes by aligning 5' expressed sequence tags (ESTs) from cap-trapped cDNA libraries to the genome, while applying stringent criteria concerning coverage and 5'-end distribution. Examination of the sequences flanking these TSSs revealed the presence of well-known core promoter motifs such as the TATA box, the initiator and the downstream promoter element (DPE). We also define, and assess the distribution of, several new motifs prevalent in core promoters, including what appears to be a variant DPE motif. Among the prevalent motifs is the DNA replication-related element DRE, recently shown to be part of the recognition site for the TBP-related factor TRF2. Our TSS set was then used to retrain the computational promoter predictor McPromoter, allowing us to improve the recognition performance to over 50% sensitivity and 40% specificity. We compare these computational results to promoter prediction in vertebrates. CONCLUSIONS: There are relatively few recognizable binding sites for previously known general transcription factors in Drosophila core promoters. However, we identified several new motifs enriched in promoter regions. We were also able to significantly improve the performance of computational TSS prediction in Drosophila. PMID- 12537578 TI - New approaches to the study of developing wood. AB - Two recent papers illustrate contrasting approaches to studying gene expression during development of the xylem, the tissue that transports water and solutes around higher plants. The two methods used, studying single cells differentiating in vitro and collecting samples from across the region around the cambium of poplar trees, have both revealed genes that have altered expression during xylem development. PMID- 12537577 TI - Systematic determination of patterns of gene expression during Drosophila embryogenesis. AB - BACKGROUND: Cell-fate specification and tissue differentiation during development are largely achieved by the regulation of gene transcription. RESULTS: As a first step to creating a comprehensive atlas of gene-expression patterns during Drosophila embryogenesis, we examined 2,179 genes by in situ hybridization to fixed Drosophila embryos. Of the genes assayed, 63.7% displayed dynamic expression patterns that were documented with 25,690 digital photomicrographs of individual embryos. The photomicrographs were annotated using controlled vocabularies for anatomical structures that are organized into a developmental hierarchy. We also generated a detailed time course of gene expression during embryogenesis using microarrays to provide an independent corroboration of the in situ hybridization results. All image, annotation and microarray data are stored in publicly available database. We found that the RNA transcripts of about 1% of genes show clear subcellular localization. Nearly all the annotated expression patterns are distinct. We present an approach for organizing the data by hierarchical clustering of annotation terms that allows us to group tissues that express similar sets of genes as well as genes displaying similar expression patterns. CONCLUSIONS: Analyzing gene-expression patterns by in situ hybridization to whole-mount embryos provides an extremely rich dataset that can be used to identify genes involved in developmental processes that have been missed by traditional genetic analysis. Systematic analysis of rigorously annotated patterns of gene expression will complement and extend the types of analyses carried out using expression microarrays. PMID- 12537579 TI - Glycan arrays for functional glycomics. AB - Interactions between carbohydrates and proteins mediate intracellular traffic, cell adhesion, cell recognition and immune system function. Two recent papers describe how arrays of oligosaccharide and polysaccharide molecules can be used to investigate these interactions more fully. PMID- 12537580 TI - RNAi hushes heterochromatin. AB - Repeated DNA elements and region-specific protein modifications combine within chromosomes to form a transcriptionally silent chromatin structure called heterochromatin. Recent work in the fission yeast Schizosaccharomyces pombe reveals that RNA is also an integral component of silent heterochromatin, providing a new perspective on how heterochromatin is organized and maintained in eukaryotic cells. PMID- 12537581 TI - The mitochondrial uncoupling proteins. AB - The uncoupling proteins (UCPs) are transporters, present in the mitochondrial inner membrane, that mediate a regulated discharge of the proton gradient that is generated by the respiratory chain. This energy-dissipatory mechanism can serve functions such as thermogenesis, maintenance of the redox balance, or reduction in the production of reactive oxygen species. Some UCP homologs may not act as true uncouplers, however, and their activity has yet to be defined. The UCPs are integral membrane proteins, each with a molecular mass of 31-34 kDa and a tripartite structure in which a region of around 100 residues is repeated three times; each repeat codes for two transmembrane segments and a long hydrophilic loop. The functional carrier unit is a homodimer. So far, 45 genes encoding members of the UCP family have been described, and they can be grouped into six families. Most of the described genes are from mammals, but UCP genes have also been found in fish, birds and plants, and there is also functional evidence to suggest their presence in fungi and protozoa. UCPs are encoded in their mature form by nuclear genes and, unlike many nuclear-encoded mitochondrial proteins, they lack a cleavable mitochondrial import signal. The information for mitochondrial targeting resides in the first loop that protrudes into the mitochondrial matrix; the second matrix loop is essential for insertion of the protein into the inner mitochondrial membrane. UCPs are regulated at both the transcriptional level and by activation and inhibition in the mitochondrion. PMID- 12537582 TI - Liver sinusoidal endothelial cells represents an important blood clearance system in pigs. AB - BACKGROUND: Numerous studies in rats and a few other mammalian species, including man, have shown that the sinusoidal cells constitute an important part of liver function. In the pig, however, which is frequently used in studies on liver transplantation and liver failure models, our knowledge about the function of hepatic sinusoidal cells is scarce. We have explored the scavenger function of pig liver sinusoidal endothelial cells (LSEC), a cell type that in other mammals performs vital elimination of an array of waste macromolecules from the circulation. RESULTS: 125I-macromolecules known to be cleared in the rat via the scavenger and mannose receptors were rapidly removed from the pig circulation, 50% of the injected dose being removed within the first 2-5 min following injection. Fluorescently labeled microbeads (2 &mgr;m in diameter) used to probe phagocytosis accumulated in Kupffer cells only, whereas fluorescently labeled soluble macromolecular ligands for the mannose and scavenger receptors were sequestered only by LSEC. Desmin-positive stellate cells accumulated no probes. Isolation of liver cells using collagenase perfusion through the portal vein, followed by various centrifugation protocols to separate the different liver cell populations yielded 280 x 107 (range 50-890 x 107) sinusoidal cells per liver (weight of liver 237.1 g (sd 43.6)). Use of specific anti-Kupffer cell- and anti desmin antibodies, combined with endocytosis of fluorescently labeled macromolecular soluble ligands indicated that the LSEC fraction contained 62 x 107 (sd 12 x 107) purified LSEC. Cultured LSEC avidly endocytosed ligands for the mannose and scavenger receptors. CONCLUSIONS: We show here for the first time that pig LSEC, similar to what has been found earlier in rat LSEC, represent an effective scavenger system for removal of macromolecular waste products from the circulation. PMID- 12537584 TI - Dynamic Medicine: the start of a new online journal. PMID- 12537583 TI - A prospective assessment of the inter-laboratory variability of biochemical markers of fibrosis (FibroTest) and activity (ActiTest) in patients with chronic liver disease. AB - BACKGROUND: Biochemical markers for liver fibrosis (FibroTest) and necroinflammatory features (ActiTest) are an alternative to liver biopsy in patients with chronic hepatitis C. Our aim was to assess the inter-laboratory variability of these tests, and their 6 components (gamma-glutamyl transpeptidase, alanine aminotransferase, alpha2-macroglobulin, haptoglobin, apolipoprotein A1, and total bilirubin) and to identify factors associated with this variability. RESULTS: Serum of 24 patients with chronic hepatitis C or severe alcoholic liver disease were prospectively recorded and analyzed in one reference center and in 8 additional laboratories. When gamma-glutamyl transpeptidase and alanine aminotransferase were expressed in international units, there was no significant difference between laboratories in the results of FibroTest or ActiTest; kappa statistics were greater than 0.50 with only 0.8% of cases (3/384) with a discordance of more than one stage. The main factor significantly associated with variability was the expression of gamma-glutamyl transpeptidase and alanine aminotransferase, as multiples of upper limit of reference values. The use of standardized method with pyridoxal phosphate reduced the variability of alanine aminotransferase expression, and standardized original Szasz method reduced the variability of gamma-glutamyl transpeptidase expression. CONCLUSIONS: The variability of FibroTest and ActiTest was acceptable without clinical consequences for the prediction of the stage of liver fibrosis and grade of activity. Standardized methods and assay calibration should be used and expression of alanine aminotransferase and gamma-glutamyl transpeptidase in multiples of the upper limit of reference values should not be employed. PMID- 12537585 TI - Genetic abnormalities in pancreatic cancer. AB - The incidence and mortality of pancreatic adenocarcinoma are nearly coincident having a five-year survival of less than 5%. Enormous advances have been made in our knowledge of the molecular alterations commonly present in ductal cancer and other pancreatic malignancies. One significant outcome of these studies is the recognition that common ductal cancers have a distinct molecular fingerprint compared to other nonductal or endocrine tumors. Ductal carcinomas typically show alteration of K-ras, p53, p16INK4, DPC4 and FHIT, while other pancreatic tumor types show different aberrations. Among those tumors arising from the exocrine pancreas, only ampullary cancers have a molecular fingerprint that may involve some of the same genes most frequently altered in common ductal cancers. Significant molecular heterogeneity also exists among pancreatic endocrine tumors. Nonfunctioning pancreatic endocrine tumors have frequent mutations in MEN 1 and may be further subdivided into two clinically relevant subgroups based on the amount of chromosomal alterations. The present review will provide a brief overview of the genetic alterations that have been identified in the various subgroups of pancreatic tumors. These results have important implications for the development of genetic screening tests, early diagnosis, and prognostic genetic markers. PMID- 12537586 TI - Implications of growth factor alterations in the treatment of pancreatic cancer. AB - Pancreatic cancer ranks fifth as a cause of cancer-related death in the world with an overall 5-year survival rate of less than 1% and a median survival of less than a year after tumour detection. Most of these patients have already metastases at the time of diagnosis. The oncologic strategies such as chemotherapy, radiotherapy, antihormonal modalities or the systemic use of specific monoclonal antibodies have not achieved a significant improvement in the survival of pancreatic cancer patients. Recent studies suggest that alterations in molecular pathways, particularly in growth factor mediated mechanisms, that regulate cell proliferation and differentiation play a pivotal role in the pathogenesis of this cancer. The molecular knowledge regarding changes in the expression of growth factors in pancreatic cancer has the potential to improve diagnostic and therapeutic treatment strategies in the near future. PMID- 12537587 TI - Gleevec (STI-571) inhibits lung cancer cell growth (A549) and potentiates the cisplatin effect in vitro. AB - BACKGROUND: Gleevec (aka STI571, Imatinib) is a recently FDA approved anti-tumor drug for chronic myelogenous leukemia. Gleevec binds specifically to BCR-ABL tyrosine kinase and inhibit the tyrosine kinase activity. It cross-reacts with another two important membrane tyrosine kinase receptors, c-kit and PDGF receptors. We sought to investigate if Gleevec has a potential role in treatment of non-small cell lung cancer. RESULTS: We have shown that Gleevec alone can inhibit the A549 lung cancer cell growth in dose-dependent manner, and the optimal concentration of Gleevec inhibition of A549 cell growth is at the range of 2-3 microM (IC50). We have also shown that A549 cells are resistant to cisplatin treatment (IC50 64 microM). Addition of Gleevec to the A549 cells treated with cisplatin resulted in a synergistic cell killing effect, suggesting that Gleevec can potentiate the effect of cisplatin on A549 cells. We also showed that the A549 lung cancer cells expresses the platelet derived growth factor receptor alpha, and the inhibitory effects of Gleevec on A549 cells is likely mediated through inhibition of PDGFR alpha phosphorylation. We further tested 33 lung cancer patients' tumor specimens to see the frequency of PDGFR-alpha expression by tissue micro-arrays and immunohistochemistry. We found that 16 of the 18 squamous carcinomas (89%), 11 of the 11 adenocarcinomas (100%), and 4 of the 4 small cell lung cancers (100%) expressed PDGFR-alpha. CONCLUSION: These results suggest a potential role of Gleevec as adjuvant therapeutic agent for treatment of non-small cell lung cancer. PMID- 12537589 TI - The application of geographical information systems to important public health problems in Africa. AB - Africa is generally held to be in crisis, and the quality of life for the majority of the continent's inhabitants has been declining in both relative and absolute terms. In addition, the majority of the world's disease burden is realised in Africa. Geographical information systems (GIS) technology, therefore, is a tool of great inherent potential for health research and management in Africa. The spatial modelling capacity offered by GIS is directly applicable to understanding the spatial variation of disease, and its relationship to environmental factors and the health care system. Whilst there have been numerous critiques of the application of GIS technology to developed world health problems it has been less clear whether the technology is both applicable and sustainable in an African setting. If the potential for GIS to contribute to health research and planning in Africa is to be properly evaluated then the technology must be applicable to the most pressing health problems in the continent. We briefly outline the work undertaken in HIV, malaria and tuberculosis (diseases of significant public health impact and contrasting modes of transmission), outline GIS trends relevant to Africa and describe some of the obstacles to the sustainable implementation of GIS. We discuss types of viable GIS applications and conclude with a discussion of the types of African health problems of particular relevance to the application of GIS. PMID- 12537588 TI - Geographical and seasonal correlation of multiple sclerosis to sporadic schizophrenia. AB - BACKGROUND: Clusters by season and locality reveal a striking epidemiological overlap between sporadic schizophrenia and multiple sclerosis (MS). As the birth excesses of those individuals who later in life develop schizophrenia mirror the seasonal distribution of Ixodid ticks, a meta analysis has been performed between all neuropsychiatric birth excesses including MS and the epidemiology of spirochaetal infectious diseases. RESULTS: The prevalence of MS and schizophrenic birth excesses entirely spares the tropical belt where human treponematoses are endemic, whereas in more temperate climates infection rates of Borrelia garinii in ticks collected from seabirds match the global geographic distribution of MS. If the seasonal fluctuations of Lyme borreliosis in Europe are taken into account, the birth excesses of MS and those of schizophrenia are nine months apart, reflecting the activity of Ixodes ricinus at the time of embryonic implantation and birth. In America, this nine months' shift between MS and schizophrenic births is also reflected by the periodicity of Borrelia burgdorferi transmitting Ixodes pacificus ticks along the West Coast and the periodicity of Ixodes scapularis along the East Coast. With respect to Ixodid tick activity, amongst the neuropsychiatric birth excesses only amyotrophic lateral sclerosis (ALS) shows a similar seasonal trend. CONCLUSION: It cannot be excluded at present that maternal infection by Borrelia burgdorferi poses a risk to the unborn. The seasonal and geographical overlap between schizophrenia, MS and neuroborreliosis rather emphasises a causal relation that derives from exposure to a flagellar virulence factor at conception and delivery. It is hoped that the pathogenic correlation of spirochaetal virulence to temperature and heat shock proteins (HSP) might encourage a new direction of research in molecular epidemiology. PMID- 12537590 TI - Antituberculosis drug resistance patterns in two regions of Turkey: a retrospective analysis. AB - BACKGROUND: The emergence of Mycobacterium tuberculosis strains resistant to antituberculosis agents has recently received increased attention owing largely to the dramatic outbreaks of multi drug resistance tuberculosis (MDR-TB). METHODS: Patients residing in Zonguldak and Kayseri provinces of Turkey with, pulmonary tuberculosis diagnosed between 1972 and 1999 were retrospectively identified. Drug susceptibility tests had been performed for isoniazid (INH), rifampin (RIF), streptomycin (SM), ethambutol (EMB) and thiacetasone (TH) after isolation by using the resistance proportion method. RESULTS: Total 3718 patients were retrospectively studied. In 1972-1981, resistance rates for to SM and INH were found to be 14.8% and 9.8% respectively (n: 2172). In 1982-1991 period, resistance rates for INH, SM, RIF, EMB and TH were 14.2%, 14.4%, 10.5%, 2.7% and 2.9% (n: 683), while in 1992-1999 period 14.4%, 21.1%, 10.6%, 2.4% and 3.7% respectively (n: 863). Resistance rates were highest for SM and INH in three periods. MDR-TB patients constituted 7.3% and 6.6% of 1982-1991 and 1992-1999 periods (p > 0.05). CONCLUSION: This study demonstrates the importance of resistance rates for TB. Continued surveillance and immediate therapeutic decisions should be undertaken in order to prevent the dissemination of such resistant strains. PMID- 12537591 TI - Economic evaluation of the benefits of reducing acute cardiorespiratory morbidity associated with air pollution. AB - BACKGROUND: Few assessments of the costs and benefits of reducing acute cardiorespiratory morbidity related to air pollution have employed a comprehensive, explicit approach to capturing the full societal value of reduced morbidity. METHODS: We used empirical data on the duration and severity of episodes of cardiorespiratory disease as inputs to complementary models of cost of treatment, lost productivity, and willingness to pay to avoid acute cardiorespiratory morbidity outcomes linked to air pollution in epidemiological studies. A Monte Carlo estimation procedure was utilized to propagate uncertainty in key inputs and model parameters. RESULTS: Valuation estimates ranged from 13 dollars (1997, Canadian) (95% confidence interval, 0-28 dollars) for avoidance of an acute respiratory symptom day to 5,200 dollars (4,000 dollars-6,400 dollars) for avoidance of a cardiac hospital admission. Cost of treatment accounted for the majority of the overall value of cardiac and respiratory hospital admissions as well as cardiac emergency department visits, while lost productivity generally represented a small proportion of overall value. Valuation estimates for days of restricted activity, asthma symptoms and acute respiratory symptoms were sensitive to alternative assumptions about level of activity restriction. As an example of the application of these values, we estimated that the observed decrease in particulate sulfate concentrations in Toronto between 1984 and 1999 resulted in annual benefits of 1.4 million dollars (95% confidence interval 0.91 1.8 million dollars) in relation to reduced emergency department visits and hospital admissions for cardiorespiratory disease. CONCLUSION: Our approach to estimating the value of avoiding a range of acute morbidity effects of air pollution addresses a number of limitations of the current literature, and is applicable to future assessments of the benefits of improving air quality. PMID- 12537592 TI - Respiratory symptoms and occupation: a cross-sectional study of the general population. AB - BACKGROUND: This study focused on respiratory symptoms due to occupational exposures in a contemporary general population cohort. Subjects were from the Dutch Monitoring Project on Risk Factors for Chronic Diseases (MORGEN). The composition of this population enabled estimation of respiratory risks due to occupation from the recent past for both men and women. METHODS: The study subjects (aged 20-59) were all inhabitants of Doetinchem, a small industrial town, and came from a survey of a random sample of 1104 persons conducted in 1993. A total of 274 cases with respiratory symptoms (subdivided in asthma and bronchitis symptoms) and 274 controls without symptoms were matched for age and sex. Relations between industry and occupation and respiratory symptoms were explored and adjusted for smoking habits and social economic status. RESULTS: Employment in the 'construction' (OR = 3.38; 95%CI 1.02 - 11.27), 'metal' (OR = 3.17; 95%CI 0. 98 - 10.28), 'rubber, plastics and synthetics' (OR = 6.52; 95%CI 1.26 - 53.80), and 'printing' industry (OR = 3.96; 95%CI 0.85 - 18.48) were positively associated with chronic bronchitis symptoms. In addition, the 'metal' industry was found to be weakly associated with asthma symptoms (OR = 2.59; 95%CI 0.87 - 7.69). Duration of employment within these industries was also positively associated with respiratory symptoms. CONCLUSION: Respiratory symptoms in the general population are traceable to employment in particular industries even in a contemporary cohort with relatively young individuals. PMID- 12537593 TI - Estimation of current density distribution under electrodes for external defibrillation. AB - BACKGROUND: Transthoracic defibrillation is the most common life-saving technique for the restoration of the heart rhythm of cardiac arrest victims. The procedure requires adequate application of large electrodes on the patient chest, to ensure low-resistance electrical contact. The current density distribution under the electrodes is non-uniform, leading to muscle contraction and pain, or risks of burning. The recent introduction of automatic external defibrillators and even wearable defibrillators, presents new demanding requirements for the structure of electrodes. METHOD AND RESULTS: Using the pseudo-elliptic differential equation of Laplace type with appropriate boundary conditions and applying finite element method modeling, electrodes of various shapes and structure were studied. The non uniformity of the current density distribution was shown to be moderately improved by adding a low resistivity layer between the metal and tissue and by a ring around the electrode perimeter. The inclusion of openings in long-term wearable electrodes additionally disturbs the current density profile. However, a number of small-size perforations may result in acceptable current density distribution. CONCLUSION: The current density distribution non-uniformity of circular electrodes is about 30% less than that of square-shaped electrodes. The use of an interface layer of intermediate resistivity, comparable to that of the underlying tissues, and a high-resistivity perimeter ring, can further improve the distribution. The inclusion of skin aeration openings disturbs the current paths, but an appropriate selection of number and size provides a reasonable compromise. PMID- 12537595 TI - A methodological report from the Malmo Diet and Cancer study: development and evaluation of altered routines in dietary data processing. AB - BACKGROUND: In the Malmo Diet and Cancer study, information on dietary habits was obtained through a modified diet history method, combining a 7-day menu book for cooked meals and a diet questionnaire for foods with low day-to-day variation. Half way through the baseline data collection, a change of interview routines was implemented in order to reduce interview time. METHODS: Changes concentrated on portion-size estimation and recipe coding of mixed dishes reported in the menu book. All method development and tests were carefully monitored, based on experiential knowledge, and supplemented with empirical data. A post hoc evaluation study using "real world" data compared observed means of selected dietary variables before and after the alteration of routines handling dietary data, controlling for potential confounders. RESULTS: These tests suggested that simplified coding rules and standard portion-sizes could be used on a limited number of foods, without distortions of the group mean nutrient intakes, or the participants' ranking. The post hoc evaluation suggested that mean intakes of energy-adjusted fat were higher after the change in routines. The impact appeared greater in women than in men. CONCLUSIONS: Future descriptive studies should consider selecting subsets assessed with either method version to avoid distortion of observed mean intakes. The impact in analytical studies may be small, because method version and diet assistant explained less than 1 percent of total variation. The distribution of cases and non-cases across method versions should be monitored. PMID- 12537596 TI - The uptake of tocopherols by RAW 264.7 macrophages. AB - BACKGROUND: Alpha-Tocopherol and gamma-tocopherol are the two major forms of vitamin E in human plasma and the primary lipid soluble antioxidants. The dietary intake of gamma-tocopherol is generally higher than that of alpha-tocopherol. However, alpha-tocopherol plasma levels are about four fold higher than those of gamma-tocopherol. Among other factors, a preferential cellular uptake of gamma tocopherol over alpha-tocopherol could contribute to the observed higher plasma alpha-tocopherol levels. In this investigation, we studied the uptake and depletion of both alpha-tocopherol and gamma-tocopherol (separately and together) in cultured RAW 264.7 macrophages. Similar studies were performed with alpha tocopheryl quinone and gamma-tocopheryl quinone, which are oxidation products of tocopherols. RESULTS: RAW 264.7 macrophages showed a greater uptake of gamma tocopherol compared to alpha-tocopherol (with uptake being defined as the net difference between tocopherol transported into the cells and loss due to catabolism and/or in vitro oxidation). Surprisingly, we also found that the presence of gamma-tocopherol promoted the cellular uptake of alpha-tocopherol. Mass balance considerations suggest that products other than quinone were formed during the incubation of tocopherols with macrophages. CONCLUSION: Our data suggests that gamma-tocopherol could play a significant role in modulating intracellular antioxidant defence mechanisms. Moreover, we found the presence of gamma-tocopherol dramatically influenced the cellular accumulation of alpha tocopherol, i.e., gamma-tocopherol promoted the accumulation of alpha-tocopherol. If these results could be extrapolated to in vivo conditions they suggest that gamma-tocopherol is selectively taken up by cells and removed from plasma more rapidly than alpha-tocopherol. This could, in part, contribute to the selective maintenance of alpha-tocopherol in plasma compared to gamma-tocopherol. PMID- 12537597 TI - Nutrition support in cancer patients: a brief review and suggestion for standard indications criteria. AB - The indications of nutrition support in cancer patients have been subject to controversy. Most studies address the effects of the method in increasing the survival or the tumor response rate. Few studies have focused on the effects in improving quality of life. After a brief review, we described the results of a study, which evaluated the effects of protein-caloric supplementation on the quality of life parameters in a group of head and neck cancer patients submitted to radiotherapy. The results support the suggestion of creating standard criteria to indicate nutrition support in cancer patients. Based on our findings, nutrition support should be indicated for cancer patients considering the potential effects to improve the quality of life. PMID- 12537594 TI - Effect of garlic on cardiovascular disorders: a review. AB - Garlic and its preparations have been widely recognized as agents for prevention and treatment of cardiovascular and other metabolic diseases, atherosclerosis, hyperlipidemia, thrombosis, hypertension and diabetes. Effectiveness of garlic in cardiovascular diseases was more encouraging in experimental studies, which prompted several clinical trials. Though many clinical trials showed a positive effect of garlic on almost all cardiovascular conditions mentioned above, however a number of negative studies have recently cast doubt on the efficacy of garlic specially its cholesterol lowering effect of garlic. It is a great challenge for scientists all over the world to make a proper use of garlic and enjoy its maximum beneficial effect as it is the cheapest way to prevent cardiovascular disease. This review has attempted to make a bridge the gap between experimental and clinical study and to discuss the possible mechanisms of such therapeutic actions of garlic. PMID- 12537598 TI - Tolerability and efficacy of single dose albendazole, diethylcarbamazine citrate (DEC) or co-administration of albendazole with DEC in the clearance of Wuchereria bancrofti in asymptomatic microfilaraemic volunteers in Pondicherry, South India: a hospital-based study. AB - BACKGROUND: The tolerability and efficacy of single dose albendazole (400 mg), diethylcarbamazine citrate (DEC) (6 mg/kg bodyweight) or co-administration of albendazole (400 mg) + DEC (6 mg/kg bodyweight) was studied in 54 asymptomatic Wuchereria bancrofti microfilaraemic volunteers in a double blind hospital-based clinical study. RESULTS: There was no significant difference in the overall incidence of adverse reactions between the three drug groups [42.1% (albendazole), 52.9% (DEC) and 61.1% (albendazole + DEC); P > 0.05]. The mean score of adverse reaction intensity did not differ significantly between the DEC and albendazole + DEC groups. However, the values in these two groups were significantly higher compared to that of albendazole alone [1.8 +/- 3.0 (albendazole) vs. 5.6 +/- 7.1 (DEC), 6.7 +/- 6.6 (albendazole + DEC); P < 0.05]. By day 360 post-therapy there was no significant difference between the three drug groups in relation to the clearance of microfilaria [26.3% (albendazole), 17.6% (DEC), 27.8% (albendazole + DEC)], reduction in geometric mean parasite density [94.7% (albendazole), 89.5% (DEC), 95.4% (albendazole + DEC)] or reduction in filarial antigenaemia [83% (albendazole), 87% (DEC), 75% (albendazole + DEC)]. Furthermore, there was a significant decrease in mean geometric parasite density (P < 0.05) as well as antigenaemia optical density values (P < 0.01) between pre-therapy levels and day 360 post-therapy in all three groups. CONCLUSIONS: This study has shown that single dose albendazole (400 mg) has similar efficacy in the clearance of microfilaria as that of DEC or the co-administration of the two drugs. The results strengthen the rationale of using albendazole for mass annual single dose administration for the control of transmission of lymphatic filariasis. PMID- 12537599 TI - MalariaSphere: a greenhouse-enclosed simulation of a natural Anopheles gambiae (Diptera: Culicidae) ecosystem in western Kenya. AB - BACKGROUND: The development and implementation of innovative vector control strategies for malaria control in Africa requires in-depth ecological studies in contained semi-field environments. This particularly applies to the development and release of genetically-engineered vectors that are refractory to Plasmodium infection. Here we describe a modified greenhouse, designed to simulate a natural Anopheles gambiae Giles ecosystem, and the first successful trials to complete the life-cycle of this mosquito vector therein. METHODS: We constructed a local house, planted crops and created breeding sites to simulate the natural ecosystem of this vector in a screen-walled greenhouse, exposed to ambient climate conditions, in western Kenya. Using three different starting points for release (blood-fed females, virgin females and males, or eggs), we allowed subsequent stages of the life-cycle to proceed under close observation until one cycle was completed. RESULTS: Completion of the life-cycle was observed in all three trials, indicating that the major life-history behaviours (mating, sugar feeding, oviposition and host seeking) occurred successfully. CONCLUSION: The system described can be used to study the behavioural ecology of laboratory-reared and wild mosquitoes, and lends itself to contained studies on the stability of transgenes, fitness effects and phenotypic characteristics of genetically engineered disease vectors. The extension of this approach, to enable continuous maintenance of successive and overlapping insect generations, should be prioritized. Semi-field systems represent a promising means to significantly enhance our understanding of the behavioural and evolutionary ecology of African malaria vectors and our ability to develop and evaluate innovative control strategies. With regard to genetically-modified mosquitoes, development of such systems is an essential prerequisite to full field releases. PMID- 12537600 TI - Bacterial autoinduction: looking outside the cell for new metabolic engineering targets. AB - Recent evidence has demonstrated that cell-to-cell signaling is a fundamental activity carried out by numerous microorganisms. A number of specialized processes are reported to be regulated by density-dependent signaling molecules including antibiotic production, bioluminescence, biofilm formation, genetic competence, sporulation, swarming motility and virulence. However, a more centralized role for quorum sensing is emerging where quorum signaling pathways overlap with stress and starvation circuits to regulate cellular adaptation to changing environmental conditions. The interplay of these phenomena is especially critical in the expression of recombinant proteins where elicitation of stress responses can dramatically impact cellular productivity. PMID- 12537601 TI - Cognitive status and behavioral problems in older hospitalized patients. AB - OBJECTIVES: (a) To determine the quantity and quality of behavioral problems in older hospitalized patients on acute care units; (b) to determine the burden of these behaviors on staff; and (c) to identify predictors of behavioral problems. METHODS: Upon admission, patients performed the Mini-Mental State Exam (MMSE), the Geriatric Depression Scale (GDS), and information was obtained on age, ethnicity, level of education, living arrangement, and psychiatric history. Two days post-admission, a clinical staff member caring for each patient, performed the Neuropsychiatric Inventory-Questionnaire (NPI-Q) to assess patients' behavioral problems and staff distress. PARTICIPANTS AND SETTING : Forty-two patients, over 60 years of age, admitted to medical and surgical units of the Veterans Affairs Hospitals in Palo Alto and San Francisco, participated. RESULTS: Twenty-three of 42 (55%) patients exhibited behavioral problems. Anxiety, depression, irritability, and agitation/aggression were the most frequently observed behaviors. The severity of the behavioral problems was significantly correlated with staff distress. Lower performance on the MMSE at admission was significantly associated with higher NPI-Q ratings. Specifically, of those cases with scores less than or equal to 27 on the MMSE, 66% had behavioral problems during hospitalization, compared to only 31% of those with scores greater than 27. CONCLUSION: Behavioral problems in older hospitalized patients appear to occur frequently, are a significant source of distress to staff, and can result in the need for psychiatric consultation. Assessment of the mental status of older adults at admission to hospital may be valuable in identifying individuals at increased risk for behavioral problems during hospitalization. PMID- 12537602 TI - Tumor necrosis factor-alpha +489G/A gene polymorphism is associated with chronic obstructive pulmonary disease. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is characterized by a chronic inflammatory process, in which the pro-inflammatory cytokine Tumor Necrosis Factor (TNF)-alpha is considered to play a role. In the present study the putative involvement of TNF-alpha gene polymorphisms in pathogenesis of COPD was studied by analysis of four TNF-alpha gene polymorphisms in a Caucasian COPD population. METHODS: TNF-alpha gene polymorphisms at positions -376G/A, -308G/A, 238G/A, and +489G/A were examined in 169 Dutch COPD patients, who had a mean forced expiratory volume in one second (FEV1) of 37 +/- 13%, and compared with a Dutch population control group of 358 subjects. RESULTS: The data showed that the TNF-alpha +489G/A genotype frequency tended to be different in COPD patients as compared to population controls, which was due to an enhanced frequency of the GA genotype. In line herewith, carriership of the minor allele was associated with enhanced risk of development of COPD (odds ratio = 1.9, p = 0.009). The other TNF alpha gene polymorphisms studied revealed no discrimination between patients and controls. No differences in the examined four TNF-alpha polymorphisms were found between subtypes of COPD, which were stratified for the presence of radiological emphysema. However, comparison of the COPD subtypes with controls showed a significant difference in the TNF-alpha +489G/A genotype in patients without radiological emphysema (chi2-test: p < 0.025 [Bonferroni adjusted]), while no differences between COPD patients with radiological emphysema and controls were observed. CONCLUSION: Based on the reported data, it is concluded that COPD, and especially a subgroup of COPD patients without radiological emphysema, is associated with TNF-alpha +489G/A gene polymorphism. PMID- 12537603 TI - The predictive role of serum and bronchoalveolar lavage cytokines and adhesion molecules for acute respiratory distress syndrome development and outcome. AB - BACKGROUND: The predictive role of many cytokines and adhesion molecules has not been studied systematically in acute respiratory distress syndrome (ARDS). METHODS: We measured prospectively tumour necrosis factor alpha (TNF-alpha), interleukin (IL)-1, soluble vascular adhesion molecule-1 (VCAM-1) and soluble intercellular adhesion molecule-1 (ICAM-1) in serum and bronchoalveolar lavage fluid (BALF) within 2 hours following admission, in 65 patients. The patients were divided into: those fulfilling the criteria for ARDS (n = 23, group A), those who were pre-ARDS and who developed ARDS within 24 hours (n = 14, group B), and those on pre-ARDS but who never developed ARDS (n = 28, group C). RESULTS: All the measured molecules were only found at higher levels in the serum of patients that died either with or without ARDS (P < 0.05 - P < 0.0001). Patients at risk exhibited a good negative predictive value (NPV) of the measured molecules for ARDS development both in their serum (89 to 95%) and BALF (86 to 92%) levels. In contrast to BALF, serum levels of IL-1 and adhesion molecules exhibited a good NPV (68 to 96%), sensitivity (60 to 88%) and survival specificity (74 to 96%) in all groups. All molecules in serum and BALF IL-1 were correlated with the APACHE II (P < 0.05 - P < 0.0001). Serum and BALF IL-1 as well as BALF TNF-alpha were negatively correlated to PaO2/FiO2 (all P < 0.05). CONCLUSIONS: The studied molecules have good NPV for ARDS development both in serum and BALF. Serum rather than BALF levels seem to be related to outcome. PMID- 12537604 TI - Insulin utilizes the PI 3-kinase pathway to inhibit SP-A gene expression in lung epithelial cells. AB - BACKGROUND: It has been proposed that high insulin levels may cause delayed lung development in the fetuses of diabetic mothers. A key event in lung development is the production of adequate amounts of pulmonary surfactant. Insulin inhibits the expression of surfactant protein A (SP-A), the major surfactant-associated protein, in lung epithelial cells. In the present study, we investigated the signal transduction pathways involved in insulin inhibition of SP-A gene expression. METHODS: H441 cells, a human lung adenocarcinoma cell line, or human fetal lung explants were incubated with or without insulin. Transcription run-on assays were used to determine SP-A gene transcription rates. Northern blot analysis was used to examine the effect of various signal transduction inhibitors on SP-A gene expression. Immunoblot analysis was used to evaluate the levels and phosphorylation states of signal transduction protein kinases. RESULTS: Insulin decreased SP-A gene transcription in human lung epithelial cells within 1 hour. Insulin did not affect p44/42 mitogen-activated protein kinase (MAPK) phosphorylation and the insulin inhibition of SP-A mRNA levels was not affected by PD98059, an inhibitor of the p44/42 MAPK pathway. In contrast, insulin increased p70 S6 kinase Thr389 phosphorylation within 15 minutes. Wortmannin or LY294002, both inhibitors of phosphatidylinositol 3-kinase (PI 3-kinase), or rapamycin, an inhibitor of the activation of p70 S6 kinase, a downstream effector in the PI 3-kinase pathway, abolished or attenuated the insulin-induced inhibition of SP-A mRNA levels. CONCLUSION: Insulin inhibition of SP-A gene expression in lung epithelial cells probably occurs via the rapamycin-sensitive PI 3-kinase signaling pathway. PMID- 12537607 TI - Editorial. PMID- 12537608 TI - Guillotine tonsillectomy: a glimpse into its history and current status in the United Kingdom. AB - Guillotine tonsillectomy was the widely practised technique of tonsillectomy in the late 19th century as it was considered a quick and reliable method of removing tonsils. It fell into disrepute in the early 20th century. This paper reviews the history of the origin of the tonsillotome and traces the various modifications over the last few centuries. The current practice of guillotine tonsillectomy is examined by means of a postal questionnaire survey of all UK consultants. PMID- 12537609 TI - Freud's friend Fliess. AB - Fliess, an ENT surgeon, was Freud's closest friend and confidant. They both believed that sexual problems were the chief cause of neurosis. Fliess pstulated that reflex nasal neurosis was based on the important physiological connection between the nose and the genitals. He described specific genital spots located on the nasal inferior turbinate. Fliess' second preoccupation was with vital periodicities. He believed that the symptoms of his reflex nasal neurosis followed regular 28-day cycles as does menstruation. He further proposed a male 23-day menstrual cycle, that he centred specifically on the nasal turbinate. Clearly, Fliess' fanciful theories of neurosis based on the turbinates have never held any scientific validity and are presented for their curiosity. This eccentric rhinolaryngologist, however, exerted a profound influence on Freud's conception of human development, that is often undervalued. PMID- 12537605 TI - Oxygen-sensing mechanisms and the regulation of redox-responsive transcription factors in development and pathophysiology. AB - How do organisms sense the amount of oxygen in the environment and respond appropriately when the level of oxygen decreases? Oxygen sensing and the molecular stratagems underlying the process have been the focus of an endless number of investigations trying to find an answer to the question: "What is the identity of the oxygen sensor?" Dynamic changes in pO2 constitute a potential signaling mechanism for the regulation of the expression and activation of reduction-oxidation (redox)-sensitive and oxygen-responsive transcription factors, apoptosis-signaling molecules and inflammatory cytokines. The transition from placental to lung-based respiration causes a relatively hyperoxic shift or oxidative stress, which the perinatal, developing lung experiences during birth. This variation in DeltapO2, in particular, differentially regulates the compartmentalization and functioning of the transcription factors hypoxia inducible factor-1alpha (HIF-1alpha) and nuclear factor-kappaB (NF-kappaB). In addition, oxygen-evoked regulation of HIF-1alpha and NF-kappaB is closely coupled with the intracellular redox state, such that modulating redox equilibrium affects their responsiveness at the molecular level (expression/transactivation). The differential regulation of HIF-1alpha and NF-kappaB in vitro is paralleled by oxygen-sensitive and redox-dependent pathways governing the regulation of these factors during the transition from placental to lung-based respiration ex utero. The birth transition period in vivo and ex utero also regulates apoptosis signaling pathways in a redox-dependent manner, consistent with NF-kappaB being transcriptionally regulated in order to play an anti-apoptotic function. An association is established between oxidative stress conditions and the augmentation of an inflammatory state in pathophysiology, regulated by the oxygen and redox-sensitive pleiotropic cytokines. PMID- 12537610 TI - Revision surgery for chronic otitis media: recurrent-residual disease and hearing. AB - The aim of this study was to determine the effect of surgical approach, intact canal wall (ICW) or canal wall down (CWD), upon the success of revision surgery for chronic otitis media (COM). A retrospective analysis of 367 patients (including 65 children aged <15 years) who underwent revision tympanoplasty because of persistent disease was performed. Single-staged tympanoplasty was performed, preserving the canal wall when present. Hearing was reconstructed with allograft incus. Follow-up ranged from one to 15 years. Hearing was determined by pre- and post-operative air-bone gaps. Post-operative re-perforation, aural discharge and/or cholesteatoma rates were similar for CWD and ICW. Cholesteatoma could present following the revision, even though it was not apparent at surgery. Following tympanoplasty, the final hearing was not significantly affected by the surgical approach or presence of cholesteatoma. Improvement in hearing was adversely affected by cholesteatoma or an absent stapes suprastructure. Revision ICW and CWD operations were both successful in controlling signs of COM. Cholesteatoma is a peripheral risk in COM and may become apparent after revision surgery. PMID- 12537611 TI - Nasal septum buttons: symptom scores and satisfaction. AB - Patients with symptomatic septal perforations require treatment, with many surgeons advocating primary treatment with an obturator if conservative measures fail. Twenty nasal Button Outcome Questionnaires were sent to patients who had undergone insertion of a septal button between 1990 and 2000 in our unit. Fourteen questinnaires were returned. This study reveals that despite a reduction in symptom score in nine patients, septal buttons are poorly tolerated by patients with only 45 per cent of obturators ultimately being in situ. In view of these findings, patients are now offered a choice between primary surgical and mechanical closure in our unit. PMID- 12537612 TI - Rationale for osteotome selection in rhinoplasty. AB - Proper selection of an osteotome for nasal osteotomy is important for minimizing soft tissue trauma. Radiographic analysis of the facial bony lateral wall thickness was performed to suggest a guideline for an appropriate osteotome size for Asians. Facial bone computed tomography (CT) of 100 patients (50 male, 50 female) were studied. The thickness of the facial bony lateral wall at three points along the track of a lateral osteotomy, and two points along the track of a medial osteotomy and intermediate osteotomy were measured. The average bony thickness along the track of a lateral osteotomy was 2.61 +/- 0.66 mm at the low level, 2.75 +/- 0.76 mm at the middle level, and 2.72 +/- 0.53 mm at the high level in subjects. The average bony thickness along the track of an intermediate osteotomy were 1.26 +/- 0.34 mm at the low level, and 1.31 +/- 0.32 mm at the high level in the subjects. The average bony thickness along the track of the medial osteotomy were 2.54 +/- 0.31 mm at the low level, and 2.77 +/- 0.30 mm at the high level in subjects. These results may provide a guideline for choosing an osteotome of appropriate size for the Asian population. PMID- 12537613 TI - Computer imaging and patient satisfaction in rhinoplasty surgery. AB - The measurement and achievement of improved patient benefit following a particular medical or surgical intervention has become an increasingly relevant part of the provision of effective healthcare. We have retrospectively analysed patient satisfaction in 56 patients following rhinoplasty via the Glasgow Benefit Inventory (GBI), 25 of whom underwent pre-operative computer imaging planning. We have also audited patient reaction to this technique via a concurrent questionnaire in those subjects who underwent imaging, and correlated this with overall patient outcome. Patient satisfaction with cosmetic rhinoplasty following computer imaging was significantly improved compared to those patients who did not receive imaging. PMID- 12537614 TI - Characterization of effective primary voice therapy for dysphonia. AB - Despite advances in the development of voice outcome measures, there is no methodology to define and quantify the elements of the complex process of speech therapy. The components of therapy given by one therapist to the intervention limb of a controlled trial were characterized according to a list of five minutes. The intervention was of proven benefit compared with a control period of observation. Indirect approaches comprised two thirds of therapy time. The types and duration of intervention were assessed but no treatment category seemed more associated with a favourable outcome. The design shows that it is possible to perform a prospective, structured analysis of the components of voice therapy. The method appears viable for the future comparison of the widely varying techniques current in voice therapy practice. PMID- 12537615 TI - Endoscopic laser treatment in pre-malignant and malignant vocal fold epithelial lesions. AB - Endoscopic laser treatment was performed in 43 patients with pre-malignant or malignant vocal fold epithelial lesions, 10 were treated with endoscopic laser surgery for dysplasia, 12 for carcinoma in situ (CIS), five for verrucous carcinoma and 16 patients for squamous cell carcinoma (SCC). Thirty-two patients received laser therapy as their first therapy, whereas 11 patients had had previous radiation therapy for laryngeal carcinoma (n = 9) or CIS (n = 2). Recurrence after initial laser therapy necessitating re-treatment (a second laser treatment or radiotherapy) occurred in nine out of 32 patients (28 per cent), thus 23 (72 per cent) were maintained free of disease during the follow-up period. Besides the 32 patients without previous therapy, patients who had already undergone radiation therapy were also included in this study. In this group there were nine patients with SCC, one patient with CIS and one with dysplasia. They all underwent laser therapy. Four were free of disease during follow-up (36 per cent) and seven developed recurrences. Six (58 per cent of all patients with previous radiation therapy) underwent total laryngectomy. PMID- 12537616 TI - Short-term complications of percutaneous tracheostomy: experience of a district general hospital--otolaryngology department. AB - Percutaneous tracheostomy is a procedure frequently carried out in a critical care setting. It is performed in the majority of cases by anaesthetists in the United Kingdom. The ENT surgeon is only called in situations where it is deemed by the intensivist that percutanous tracheostomy would prove too great a risk. In this situation the patient was taken to the operating theatre for a surgical tracheostomy. In our paper, a retrospective analysis was performed of all percutaneous tracheostomies carried out by ENT surgeons in the Royal Glamorgan Hospital, during a two-year period from July 1999 to July 2001, to assess whether percutaneous tracheostomy is a feasible option as a first line procedure in all elective tracheostomies. Thirty-six patients were included in the study. The mean age was 60.2 years. Haemorrhage was noted to be a problem in only one patient and two patients developed postoperative wound infection that was treated with systemic antibiotics. No other complications were encountered. We propose that all ENT surgeons should be trained in performing percutaneous tracheostomy and that it should be used as the gold standard in elective tracheostomy insertion. In cases where difficulties are likely to be anticipated, percutaneous tracheostomy can still be considered as the first option. This can be performed in the operating theatre setting with the knowledge that if any complication should occur then conversion to surgical tracheostomy can be done without delay. PMID- 12537617 TI - Learning from patients with cancer and their spouses: a focus group study. AB - Guidelines, standards and protocols are now widespread in clinical oncology. Although based on randomized trial evidence where possible, most are based on professional consensus. However, the goal of incorporating patient and carer views meaningfully in such guidelines has proved elusive. We performed a qualitative, focus group study of patients with head and neck cancer, their carers and bereaved carers in the South and West of England. Patients and carers were asked to discuss their views on a series of professionally-derived standards for care. Pilot work showed that convening patient groups twice was more productive, allowing the first to discuss 'the patient story/journey' and the second to be more directed at specific standards, suitably translated into lay language. The results demonstrated that such methodology was applicable even to groups of patients traditionally viewed as 'difficult', such as laryngectomy patients, facially disfigured persons and bereaved carers. Participants were able to comment meaningfully on the standards and the process of head and neck cancer care as a whole. National (BAO-HNS) standards were modified with the results of the study. Focus groups, especially reconvened groups, are an effective way of gaining patient and carer views of professionally-derived standards in oncology. However, professional qualitative researcher help is required, and it is not necessarily 'easy' or cheap. PMID- 12537618 TI - A case of rhinolithiasis in botswana: a mineralogical, microscopic and chemical study. AB - A case of rhinolithiasis in Southeast Botswana was treated and after removal in hospital, the rhinolith was subjected to macroscopic and microscopic examination, X-ray diffraction analysis, electron microscope analysis and partial botanical analysis. The rhinolith consists of a strongly elliptical core of calcium stearate (C36H70CaO4.H2O), surrounded by approximately 30 elongated concentric growth rings, consisting of sodium-containing whitlockite (Ca18Mg2(Na,H)(PO4)14). The different layers have various degrees of porosity and red staining, probably due to traces of amorphous iron oxide. The origin of the rhinolith started with a piece of plant material, lodged in the nose, which was replaced by calcium stearate, leaving some remnants of resistant epidermal plant tissue. During subsequent years, thin layers of whitlockite were deposited periodically around the core with the reddish brown bands representing deposition during the dry season when atmospheric dust rich in amorphous iron oxide is at its highest in Botswana. PMID- 12537619 TI - Cholesterol granuloma of the frontal sinus. AB - It is common to see cholesterol granuloma in the mastoid air cells, less common in the orbit, and uncommon in the paranasal sinuses. Cholesterol granuloma is thought to be due to an interruption to normal aeration with impaired lymphatic drainage, resulting in a closed cavity where it may form. These expanding cysts cause bone destruction and compression of the surrounding structures that lead to clinical symptoms. Diagnosis and management of cholesterol granuloma cysts can be challenging. Magnetic resonance imaging (MRI) and computed tomographic (CT) scans are usually diagnostic. We present a rare case of cholesterol granuloma in the frontal sinus, few cases have been reported in the literature. PMID- 12537620 TI - Metachronous adenocarcinoma of the maxillary sinuses: a rare entity. AB - Malignant tumours of the nasal cavities and paranasal sinuses are uncommon. They constitute less than one per cent of all tumours and less than three per cent of head and neck tumours. Although multiple primary carcinomas of the aerodigestive tract are commonly reported, metachronous maxillary sinus carcinomas are rare. To date, all of these cases reported are of squamous cell origin. The incidence of metachronous maxillary carcinoma involving both sinuses is in the range of 1.4 per cent amongst patients with maxillary carcinoma. We present the first reported case, to our knowledge, of a patient with metachronous adenocarcinoma of the maxillary sinus. PMID- 12537621 TI - Familial vocal fold paralysis. AB - Familial clustering of congenital bilateral abductor vocal fold paralysis has been reported very rarely. So far, only a handful of cases have been reported, mostly with the autosomal dominant of X-linked recessive mode of inheritance. We describe the cases of a brother and sister, who presented with neonatal stridor due to bilateral abductor vocal fold paralysis. First-degree parental consanguinity suggests an autosomal recessive mode of inheritance. Karyotype analysis revealed a paracentric balanced inversion of chromosome 13 in both cases, that was also present in the unaffected mother. An updated review of the literature on this interesting but rare condition is also presented. PMID- 12537622 TI - A solitary neurofibroma of the palatine tonsil. AB - A rare case of a tonsillar neurofibroma in a 28-year-old Caucasian male is presented. Benign tumours of the tonsils are rare and of those reported, only a few are benign peripheral nerve sheath tumours (BPNSTs). This is the first report of a solitary neurofibroma of the palatine tonsil in the English literature. PMID- 12537623 TI - Tongue ischaemia in a patient with external carotid artery stenosis. AB - We report the case of a 75-year-old man who presented with an ischaemic tongue. He was known to have external carotid artery stenosis and a history of a transient ischaemic attack. He was treated with a heparin infusion and the tongue healed well. PMID- 12537624 TI - A base of skull fracture presenting as haematemesis. AB - In a patient presenting with haematemesis, known to have oesophageal varices, the bleeding would be expected to be from those varices. This case demonstrates an unusual presentation of a base of skull fracture; blood from a fracture of the temporal bone had run from the middle ear through the eustachian tube to the pharynx. It demonstrates the need to remain vigilant even when the diagnosis appears to be obvious. PMID- 12537625 TI - Imrich Friedmann (1907-2002). PMID- 12537626 TI - Acridine Orange for malaria diagnosis: its diagnostic performance, its promotion and implementation in Tanzania, and the implications for malaria control. AB - One hundred years ago, Giemsa's stain was employed for the first time for malaria diagnosis. Giemsa staining continues to be the method of choice in most malarious countries, although, in the recent past, several alternatives have been developed that exhibit some advantages. Considerable progress has been made with fluorescent dyes, particularly with Acridine Orange (AO). The literature on the discovery, development and validation of the AO method for malaria diagnosis is reviewed here. Compared with conventional Giemsa staining, AO shows a good diagnostic performance, with sensitivities of 81.3%-100% and specificities of 86.4%-100%. However, sensitivities decrease with lower parasite densities, and species differentiation may occasionally be difficult. The most notable advantage of the AO method over Giemsa staining is its promptness; results are readily available within 3-10 min, whereas Giemsa staining may take 45 min or even longer. This is an important advantage for the organization of health services and the provision of effective treatment of malaria cases. The national malaria control programme of Tanzania, together with the Japan International Co-operation Agency, began to introduce the AO method in Tanzania in 1994. So far, AO staining has been introduced in 70 regional and district hospitals, and 400 laboratory technicians have been trained to use the method. The results of this introduction, which are reviewed here and have several important implications, indicate that AO is a viable alternative technique for the laboratory diagnosis of malaria in highly endemic countries. PMID- 12537627 TI - Chloroquine or sulfadoxine-pyrimethamine for the treatment of uncomplicated, Plasmodium falciparum malaria during an epidemic in Central Java, Indonesia. AB - A recent malaria epidemic in the Menoreh Hills of Central Java has increased concern about the re-emergence of endemic malaria on Java, which threatens the island's 120 million residents. A 28-day, in-vivo test of the efficacy of treatment of malaria with antimalarial drugs was conducted among 167 villagers in the Menoreh Hills. The treatments investigated, chloroquine (CQ) and sulfadoxine pyrimethamine (SP), constitute, respectively, the first- and second-line treatments for uncomplicated malaria in Indonesia. The prevalence of malaria among 1389 residents screened prior to enrollment was 33%. Treatment outcomes were assessed by microscopical diagnoses, PCR-based confirmation of the diagnoses, measurement of the whole-blood concentrations of CQ and desethylchloroquine (DCQ), and identification of the Plasmodium falciparum genotypes. The 28-day cumulative incidences of therapeutic failure for CQ and SP were, respectively, 47% (N = 36) and 22% (N = 50) in the treatment of P. falciparum, and 18% (N = 77) and 67% (N = 6) in the treatment of P. vivax. Chloroquine was thus an ineffective therapy for P. falciparum malaria, and the presence of CQ-resistant P. vivax and SP-resistant P. falciparum will further compromise efforts to control resurgent malaria on Java. PMID- 12537628 TI - The high sensitivity of a PCR-ELISA in the diagnosis of cutaneous and visceral leishmaniasis caused by Leishmania infantum. AB - In general, the conventional techniques available for the diagnosis of leishmaniasis have relatively low sensitivity. This means that parasite-rich samples (which can usually only be collected by very invasive methods, such as bone-marrow aspiration) must be employed. This problem has not yet been solved even by use of the PCR-based techniques currently available. However, a new PCR ELISA has been developed for the diagnosis of cutaneous (CL) and visceral leishmaniasis (VL) caused by Leishmania infantum. This assay appears to have sufficient sensitivity to be effective in the diagnosis of VL not only when bone marrow aspirates are investigated but also when the samples are of peripheral blood. Overall, the ability of the PCR-ELISA to detect Leishmania, in 76 samples (22 of peripheral blood, 36 bone-marrow aspirates and 18 skin samples) from 72 patients living in a endemic region, was better than that of culture or the examination of Giemsa-stained smears. For example, L. infantum kDNA was detected by PCR-ELISA in 15 (83%) of the 18 skin samples from suspected cases of CL, whereas the combined use of several classical techniques only confirmed the presence of amastigotes in five (28%) of these samples. Similarly, only 21 individuals were diagnosed as having VL by conventional techniques whereas 30 were found Leishmania-positive in the PCR-ELISA. The new PCR-ELISA also appears to be a suitable technique for detecting leishmanial kDNA in samples of peripheral blood from cases of L. infantum-HIV co-infection. The assay is more sensitive than the combined use of several conventional techniques in the diagnosis of subclinical VL, probably because those with subclinical infection have relatively low parasitic loads that are generally undetectable using the other techniques. PMID- 12537629 TI - Evaluation of the patterns of Schistosoma mansoni infection and re-infection in Senegal, from faecal egg counts and serum concentrations of circulating anodic antigen. AB - Infection and re-infection patterns were evaluated in a recent Schistosoma mansoni focus in northern Senegal, by determining concentrations of serum circulating anodic antigen (CAA), as a measure of worm burden, and counting eggs in faeces before, 6 or 12 weeks and 1 year after praziquantel treatment in two subsequent cohorts (cohort A and B). No differences in egg counts and CAA concentrations or their relationship were found between the cohorts, which were examined 2 years apart. Within both cohorts, CAA concentrations showed the same, typical, age-related patterns as egg counts, with a peak in children and a strong decline in adults. These trends were apparent both before and 1 year after treatment. The results indicate that an age-related resistance to infection and to re-infection has been firmly established, at a steady level, in the recent S. mansoni focus investigated, with no indication of a gradual development of immunity or anti-fecundity immunity over a period of 2 years. Both shortly and 1 year after treatment, the decrease in egg counts was stronger than that in CAA concentrations, indicating that that there had been a reduction in worm fecundity after treatment. The possibility that praziquantel may induce anti-fecundity immunity has important implications for the use and interpretation of the results of (egg-count-based) re-infection studies designed to follow the development of naturally acquired immunity. PMID- 12537630 TI - Rapid dot-ELISA for the detection of specific antigens in the cyst fluid from human cases of cystic echinococcosis. AB - A rapid dot-ELISA was developed for the detection of specific antigens in samples of cyst fluid from human cases of Echinococcus granulosus infection, permitting the confirmation of cystic echinococcosis (CE). Peroxidase-conjugated antibodies against antigen B (derived from the fluid in the cysts of sheep infected with E. granulosus) were used to test cyst-fluid samples from 22, surgically confirmed cases of human CE and 21 domestic animals (horses, sheep, buffalo, a cow and a camel) with CE. All of these samples were found to be strongly positive in the dot-ELISA, by direct reading with the naked eye. In contrast, fluid samples from seven, non-parasitic liver cysts of human origin were all negative, and a sample taken from the residual cavity left in a patient after previous CE surgery showed a weakly positive reaction. The antigen-detection assay, which can be completed within 10 min, may be a useful 'bedside' test for the differential diagnosis of cysts during surgery or percutaneous treatment for suspected CE. PMID- 12537631 TI - The use of spatial analysis in mapping the distribution of bancroftian filariasis in four West African countries. AB - The geographical distribution of human infection with Wuchereria bancrofti was investigated in four West African countries (Benin, Burkina Faso, Ghana and Togo), using a commercial immunochromatographic test for filarial antigen. Efforts were made to cover each health-system implementation unit and to ensure no sampling point was >50 km from another, but otherwise the 401 study communities were selected at random. The aim was to enable spatial analysis of the data, to provide a prediction of the overall spatial relationships of the infection. The results, which were subjected to an independent random validation in Burkina Faso and Ghana, revealed that prevalence in the adult population of some communities exceeded 70% and that, over large areas of Burkina Faso, community prevalences were between 30% and 50%. Most of Togo, southern Benin and much of southern Ghana appeared completely free of the infection. Although there were foci on the Ghanaian coast with prevalences of 10%-30%, such high prevalences did not extend into coastal Togo or costal Benin. The prevalence map produced should be useful in prioritizing areas for filariasis control, identifying potential overlap with ivermectin-distribution activities undertaken by onchocerciasis-control programmes, and enabling inter-country and sub-regional planning to be initiated. The results indicate that bancroftian filariasis is more widely distributed in arid areas of Burkina Faso than hitherto recognized and that the prevalences of infection have remained fairly stable for at least 30 years. The campaign to eliminate lymphatic filariasis as a public-health problem in Africa will require significantly more resources (human, financial, and logistic) than previously anticipated. PMID- 12537632 TI - Effects of a 3-day regimen of albendazole (800 mg daily) on Loa loa microfilaraemia. AB - The encephalopathy that sometimes develops after ivermectin treatment in patients with high Loa microfilaraemias is probably related to a massive effect of the drug on the Loa microfilariae. A trial was therefore conducted to evaluate whether a course of albendazole would bring about a slower decrease in the Loa microfilaraemia, and thus could be used as a mass 'clearing' treatment, before the distribution of ivermectin in areas where onchocerciasis and loiasis are co endemic. The Loa microfilarial loads were followed monthly for 9 months in two groups of subjects, one treated with albendazole (400 mg twice a day for 3 days), and the other with vitamin (B(1), B(6) and B(12)) tablets. There were no significant between-group differences in the microfilarial loads at any of the examination rounds. During the follow-up period, there was also no significant change in the overall loads among those treated with albendazole, although the counts in those with high initial microfilaraemias (>8000 microfilariae/ml) tended to decrease progressively during the first 3 months. Further trials should now be performed, to evaluate the effects on Loa loa of two courses of albendazole given 2-3 months apart. PMID- 12537633 TI - Soil-transmitted nematode infections and mebendazole treatment in Mafia Island schoolchildren. AB - In August 2000, a cross-sectional study was performed to assess the prevalence and intensity of soil-transmitted nematode infections in schoolchildren on Mafia Island. Hookworm infection was widespread (72.5% prevalence) whereas Trichuris trichiura was less prevalent (39.7%) and Ascaris lumbricoides was present at a low prevalence (4.2%), mainly in urban areas. In a subsample of the study population, both Necator americanus and Ancylostoma duodenale were found, although N. americanus was more prevalent. This survey was followed by a parasitological evaluation of mebendazole treatment using a single, 500-mg dose. The data on outcome were used for comparison with those from recent studies of similar treatment regimens in the neighbouring island of Pemba, Zanzibar, where periodic chemotherapy with mebendazole to schoolchildren has been implemented as part of a helminth-control programme since 1994. A higher efficacy of mebendazole against hookworm infection was found in Mafia Island (where a cure 'rate' of 31.3% and an egg-reduction 'rate' of 78.1% were recorded) when compared with that observed in Pemba Island, possibly indicating that hookworms may be developing mebendazole resistance on Pemba Island as a result of intense exposure to the drug there. PMID- 12537634 TI - Molecular characterization of hepatitis-A-virus infections, in the context of two outbreaks in southern Thailand. AB - As hepatitis A virus (HAV) is usually transmitted through the faecal-oral route, hepatitis A is a communicable disease. In countries of intermediate to low endemicity, sudden outbreaks of human infection with the virus may occur. Between September 2001 and April 2002, there were two outbreaks of HAV infection in the Ruso and Yeengor districts of Narathiwas province, in southern Thailand. Isolates of HAV were recovered during these outbreaks, from 14 in-patients with acute hepatitis in Ruso (12 positive for anti-HAV IgM and all positive for HAV RNA), 16 children with asymptomatic infection in Yeengor (14 positive for anti-HAV IgM and nine for HAV RNA), and four isolated cases in Bangkok (all positive for anti-HAV IgM). Molecular characterization of the VP1-P2A region of each isolate was followed by phylogenetic analysis. All of the isolates from Narathiwas province were found to be of genotype 1a, to have the same VP1 nucleotide sequence, and to show a high level of sequence homology (>/= 99.5%) with the isolates from Bangkok and with previous Thai isolates. These results should facilitate further research into HAV transmission and genotype identification in community outbreaks. PMID- 12537635 TI - Immunity to the sexual stages of Plasmodium falciparum in mothers, neonates and infants subject to intense and perennial malarial transmission. PMID- 12537636 TI - Rainy-season prevalence of malaria in Bobonaro district, East Timor. PMID- 12537637 TI - Secondary renal involvement in human cystic echinococcosis: a review of the literature and report of another case. PMID- 12537639 TI - Biography of Giorgio Cantelli Forti. PMID- 12537640 TI - Scrutinizing three trichloroethylene carcinogenicity classifications in the European Union--implications for the risk assessment process. AB - In this paper, carcinogenicity classifications of the chlorinated solvent trichloroethylene (TCE) made by the European Union Commission Expert Groups in 1976, 1988, and 2001 are scrutinized and alternative classifications are proposed. It is argued that the TCE database at these three points in time could have been interpreted to fulfill the criteria for stricter classifications than those actually made. Implications of this for the classification process are discussed. PMID- 12537641 TI - Retinal pigment epithelium melanin and ocular toxicity. AB - Ocular morphology is specifically directed at facilitating the transmittance of visible light to the retina for the purposes of photoreceptor absorption and phototransduction, thereby initiating the process of vision. By absorbing excess radiation, melanin significantly enhances this process. It can also act as a photoprotector by quenching reactive oxygen species and other radicals produced as a result of the high oxygen dependency of the retina for its metabolism. However, melanin also binds numerous pharmaceuticals, a process that can result in ocular toxicity. Although our understanding of this binding remains somewhat limited, melanin chemistry, its distribution, and other factors influencing binding appear to play a significant role in predisposing ocular tissues, such as the choroid and retinal pigment epithelium, to toxicological insult. Many of the drugs that have been identified as causing these effects are known photosensitizers in which radiation plays a significant role in eliciting the pathologies. The phototoxic effects range from minor histological/chemical changes, which do not impact the quality of vision, to pigmentary retinopathies, which could potentially involve the loss of sight. Such effects, resulting from photosensitizer-drug binding to melanin, are to be separated from toxic effects, such as some ganglion cell abnormalities, that result from pharmaceuticals affecting ocular tissues directly. PMID- 12537642 TI - Photokeratitis and other phototoxic effects on the cornea and conjunctiva. AB - Except when sleeping, the cornea and interpalpebral conjunctiva are exposed to the ambient environment, both natural and man-made. Levels of solar ultraviolet irradiance reaching the eye may exceed the damage threshold under a number of circumstances. The consequences of overexposure may be acute after a latent period, sequelae to an acute exposure, or long-term chronic effects. Previously derived action spectra for photokeratitis and photoconjunctivitis due to incoherent ultraviolet are presented. These reveal interspecies similarities for the levels of radiant energy reaching each tissue. The initial in vivo (clinical) signs of photokeratitis are due to lost or damaged epithelial cells with other signs produced by this primary response. The conjunctival signs include injection and chemosis. Chronic exposure to solar ultraviolet is a factor in climatic droplet keratopathy and pterygium. Phototoxic compounds or their by-products potentially can reach the cornea from the air, via the tears or aqueous humor, or from the limbal capillaries. However, the human cornea appears to be much less susceptible to the influence of phototoxic agents than the skin. PMID- 12537643 TI - Uveal melanocytes, ocular pigment epithelium, and Muller cells in culture: in vitro toxicology. AB - Uveal melanocytes and the ocular pigment epithelium are located in the middle and inner layers of the eye. Muller cells (a type of glial cell) are located in the neural retina. Melanocytes, retinal pigment epithelium (RPE), and Muller cells do not participate directly in the detection or transfer of visual information, but they have various functions that support the neural retina and are essential for the maintenance of vision. Methods for the isolation and cultivation of melanocytes, RPE, and Muller cells have been established by us and other investigators. These cultured cells can be used as in vitro model systems for studying the toxicology of visible light, ultraviolet (UV) radiation, drugs, and other potentially toxic agents. Toxic effects on these cells may give rise to altered retinal function and result in impaired vision. Both melanocytes and pigment epithelium contain melanin, which has the ability to bind organic amines and metal ions. This results in the accumulation of these substances in the eye. Melanin may protect cells from chemical stress by binding toxic chemicals; but in chronic exposure, increased and lengthy binding may cause damage to these cells. Two different types of melanin are found in the eye: eumelanin and pheomelanin, which may have photoprotective and phototoxic effects, respectively. Pigment epithelium contains mainly eumelanin, whereas melanocytes contain both eumelanin and pheomelanin. Melanin is an antioxidant and with age, the antioxidant properties may diminish to the point that it may even become a prooxidant. There are also other functions of pigment epithelium and uveal melanocytes not related to melanin and there are also several functions of Muller cells that play a role in the toxicological aspects of the eye. Cultured uveal melanocytes, pigment epithelial cells, and Muller cells can be used to study the toxicology of these cells in vitro. PMID- 12537644 TI - Phototoxicity to the retina: mechanisms of damage. AB - Light damage to the retina occurs through three general mechanisms involving thermal, mechanical, or photochemical effects. The particular mechanism activated depends on the wavelength and exposure duration of the injuring light. The transitions between the various light damage mechanism may overlap to some extent. Energy confinement is a key concept in understanding or predicting the type of damage mechanism produced by a given light exposure. As light energy (either from a laser or an incoherent source) is deposited in the retina, its penetration through, and its absorption in, various tissue compartments is determined by its wavelength. Strongly absorbing tissue components will tend to "concentrate" the light energy. The effect of absorbed light energy largely depends on the rate of energy deposition, which is correlated with the exposure duration. If the rate of energy deposition is too low to produce an appreciable temperature increase in the tissue, then any resulting tissue damage necessarily occurs because of chemical (oxidative) reactions induced by absorption of energetic photons (photochemical damage). If the rate of energy deposition is faster than the rate of thermal diffusion (thermal confinement), then the temperature of the exposed tissue rises. If a critical temperature is reached (typically about 10 degrees C above basal), then thermal damage occurs. If the light energy is deposited faster than mechanical relaxation can occur (stress confinement), then a thermoelastic pressure wave is produced, and tissue is disrupted by shear forces or by cavitation-nonlinear effects. Very recent evidence suggests that ultrashort laser pulses can produce tissue damage through nonlinear and photochemical mechanisms; the latter because of two-photon excitation of cellular chromophores. In addition to tissue damage caused directly by light absorption, light toxicity can be produced by the presence of photosensitizing agents. Drugs excited to reactive states by ultraviolet (UV) or visible light produce damage by type I (free radical) and type II (oxygen dependent) mechanisms. Some commonly used drugs, such as certain antibiotics, nonsteroidal anti-inflammatory drugs (NSAIDs), and psychotherapeutic agents, as well as some popular herbal medicines, can produce ocular phototoxicity. Specific cellular effects and damage end points characteristic of light damage mechanisms are described. PMID- 12537645 TI - Screening for ocular phototoxicity. AB - Normally light transmission through the eye is benign and serves to direct vision and circadian rhythm. However, with very intense light exposure, or with ambient light exposure to the aged eye and/or young or adult eye in the presence of light activated (photosensitizing) drugs or dietary supplements, cosmetics, or diagnostic dyes, light can be hazardous, leading to blinding disorders. Light damage to the human eye is avoided because the eye is protected by a very efficient antioxidant system and the chromophores present absorb light and dissipate its energy. After middle age, there is a decrease in the production of antioxidants and antioxidant enzymes and an accumulation of endogenous chromophores that are phototoxic. The extent to which a particular exogenous photosensitizing substance is capable of producing phototoxic side effects in the eye depends on several parameters, including (1) the chemical structure; (2) the absorption spectra of the drug; (3) binding of the drug to ocular tissue (lens proteins, melanin, DNA); and (4) the ability to cross blood-ocular barriers (amphiphilic or lipophilic). For instance, compounds that have either a tricyclic, heterocyclic, or porphyrin ring structure and are incorporated into ocular tissues are potentially phototoxic agents in the eye. The extent to which these substances might damage the eye (photoefficiency) can be predicted using in vitro and photophysical techniques. With simple, inexpensive testing, compounds can be screened for their potential ocular phototoxicity at the developmental stage. It may be that a portion of the molecule can be modified to reduce phototoxicity while leaving the primary drug effect intact. Preclinical safety testing may prevent ocular side effects that can range from mild, reversible blurred vision to permanent blindness. PMID- 12537646 TI - How light reaches the eye and its components. AB - The human eye is exquisitely sensitive to light (i.e., visible radiant energy), and when dark-adapted, the retina can detect a few photons of blue-green light. It is therefore not at all surprising that ocular tissues are also more vulnerable to ultraviolet (UV) and light damage than the skin. For this reason, humans have evolved with certain anatomical, physiological, and behavioral traits that protect this critical organ from the UV damage that would otherwise be certain from the intense bath of overhead solar ultraviolet radiation (UVR) when we are outdoors during daylight. For example, the UV exposure threshold dose for photokeratitis ("welders' flash" or "snow blindness")--if measured as falling on a horizontal ground surface--would be reached in less than 10 minutes around midday in the summer sun. There are three critical ocular structures that could be affected by UV exposure: the cornea, the lens, and the retina. The cornea transmits radiant energy only at 295 nm and above. The crystalline lens absorbs almost all incident energy to wavelengths of nearly 400 nm. In youth, a very small amount of UV-A reaches the retina, but the lens becomes more absorbing with age. Thus there are intraocular filters that effectively filter different parts of the UV spectrum and allow only of the order of 1% or less to actually reach the retina. Nevertheless, this small fraction of energy--if phototoxic--could still be of concern. Finally, oblique rays entering the eye from the temporal side, can actually reach the equatorial (germinative) area of the lens. PMID- 12537647 TI - The READIT assay as a method for genotyping NAT1*10 polymorphisms. AB - Polymorphisms in the N-acetyltransferases (NATs) have been associated with increased risks for the development of a variety of cancers. The NAT1*10 allele, for example, has been reported to be associated with an increased risk of colon and urinary bladder cancers, among others. Therefore, considerable effort is being placed on the development of genotyping methodologies for NAT activities both for pharmacological as well as disease preventative applications. Most NAT genotyping approaches are gel based and consist of PCR-restriction fragment length polymorphism (RFLP) analysis, allele-specific PCR, or both. Although these approaches have their utility, they are slow, labor intensive, and are not amenable to automation. Recently, a novel approach to genotyping known as the READIT Assay has been introduced. The READIT methodology involves a reversal of the DNA polymerase reaction to generate dNTPs through the phosphorolytic cleavage of oligonucleotide probe molecules annealed to target DNAs. In a coupled reaction, kinase converts the resulting dNTPs to ATP. ATP production is then monitored by the addition of luciferase, generating a light signal proportional to the amount of dNTPs generated through probe depolymerization. We describe the development of a READIT genotyping protocol for the analysis of NATs using the NAT1*10 allele as a model system and demonstrate its utility for the analysis of archival dried blood specimens. We applied this technology to genotype 678 DNAs at the NAT-1088T --> A polymorphic site, and 680 DNAs for the 1095C --> A polymorphism. We report complete concordance for the 1088T --> A polymorphism for all 678 genotypes previously determined by RFLP analysis. PMID- 12537648 TI - Rapid and robust screening of the Menkes disease/occipital horn syndrome gene. AB - Menkes disease and occipital horn syndrome (OHS) are allelic neurogenetic disorders of copper transport associated with mutations in an X-linked gene, ATP7A. This gene encodes a copper-transporting P-type ATPase. The spectrum of mutations at the Menkes/OHS locus is estimated to include 1% chromosomal rearrangements and 15-20% large deletions, with the remaining defects involving small alterations. There is a compelling need for a rapid and reliable molecular diagnostic approach for patients and families impacted by these conditions. In addition to testing suspected affected males, carrier screening of females in Menkes/OHS families and prenatal evaluation of at-risk pregnancies will be enhanced by the wider availability of robust mutation analysis for this large (23 exon) locus. Here we describe a stepwise approach to mutation screening for these disorders that successfully identified molecular alterations in over 95% of our patient population (n = 49). This genomic DNA-based technique employs multiplex PCR, heteroduplex analysis, and direct sequencing, in a serial fashion. This approach should find application in molecular diagnostic laboratories in the United States and other countries. Currently, only a single European center provides commercial testing for unknown mutations in Menkes/OHS patients, even though these disorders occur worldwide. PMID- 12537649 TI - Single-nucleotide polymorphisms and haplotype analysis in beta-defensin genes in different ethnic populations. AB - Beta-defensins are cationic antimicrobial peptides expressed by epithelial cells and exhibit antibacterial, antifungal, and antiviral properties. The defensins are part of the innate host defense network and may have a significant protective role in the oral cavity and other mucosa. Defects or alteration in expression of the beta-defensins may be associated with susceptibility to infection and mucosal disorders. We examined the occurrence of single-nucleotide polymorphisms (SNPs) in the human beta-defensin genes DEFB1 and DEFB2 encoding human beta-defensin-1 and -2 (hBD-1, hBD-2), respectively, in five ethnic populations and defined haplotypes in these populations. Fifteen SNPs were identified in both DEFB1 and DEFB2. Coding region SNPs were found in very low frequency in both genes. One nonsynonymous DEFB1 SNP, G1654A (Val --> Ile), and one nonsynonymous DEFB2 SNP, T2312A (Leu --> His), were identified. Seven sites in each gene exhibited statistically significant differences in frequency between ethnic groups, with the greatest variation in the promoter and in the 5'-untranslated region of DEFB1. DEFB1 displayed 10 common haplotypes, including one cosmopolitan haplotype. Eight common haplotypes were found in DEFB2, including one cosmopolitan haplotype shared among all five ethnic groups. Our results show that genotypic variability among ethnic groups will need to be addressed when performing associative genetic studies of innate defense mechanisms and susceptibility to disease. PMID- 12537650 TI - Editor's comment: colon cancer carrier screening enters the genetic testing arena. PMID- 12537651 TI - Attitudes toward genetic testing in patients at risk for HNPCC/FAP and the German population. AB - Adequate knowledge regarding hereditary diseases and genetics, as well as personal attitudes toward gene tests, are major determinants of optimal utilization of genetic testing. In the present study, we aimed to explore the general attitudes toward genetic testing in a sample representative of the German general population (n = 2,076) and to compare the attitudes of persons at risk for hereditary non-polyposis colorectal cancer/familial adenomatous polyposis (HNPCC/FAP) (n = 36) who had attended a university genetic counseling service, with a matched general population sample. We administered a subset of a questionnaire previously used in a Finnish study (Jallinoja et al., 1998). The 12 statements pertain to approval, disapproval, and concern for genetic testing. Overall, the results reveal high approval of genetic testing in the German population and in at-risk persons. In accordance with other studies, we find that the attitudes of individuals for whom hereditary disease is a salient issue of personal relevance and the attitudes of the general public are very similar. Only a few significant differences between these two samples emerged, indicating that at-risk persons hold a more favourable view of the testing. One intriguing finding was the high rate of "don't know" responses, especially in the general population sample. Compared to results from Finland, approval of genetic testing is lower in the German population, and endorsement of "don't knows" is remarkably higher. We argue for increased attention to the issue of attitude change after genetic counseling and for the need of comparative cross-cultural research on attitudes toward gene technology. PMID- 12537652 TI - Patient vs. physician as the target of educational outreach about screening for an inherited susceptibility to colorectal cancer. AB - Are patients identified from a cancer registry better educated directly or via their physician about screening for an inherited susceptibility for colorectal cancer? Of 974 patients diagnosed with colorectal cancer at < or = 60 years from 1987 to 1999 in a five-county area including Rochester, the physicians of 651 patients (67%) forwarded a cancer family history survey to their patient; 459 (71%) completed the survey. Of these 459, 167 (36%) reported having at least one first- or second-degree relative with colon cancer and were sent a set of questionnaires and a more detailed family cancer history form. Of the 167, a total of 101 (60%) continued to qualify by returning the questionnaires. These 101 qualifying patients were randomized to either the patient-education or physician-education group. Of the 101, a total of 47 (47%) came for a free genetic evaluation. Individuals were more likely to accept evaluation if they were parents (p = 0.001), had more cancers of all kinds in their families (p = 0.02), and had a larger social network (p = 0.04). Of the 47 counseled, 36 (77%) chose to have DNA testing at no cost. Of these 47, individuals were more likely to choose DNA testing if they had more cancers in the family (p = 0.04) and fewer symptoms of depression (p = 0.05). Of the 36 tested patients, 6 (20%) were found to have mutations. In summary, acceptance of genetic services was related to the magnitude of the threat (more cancers in the family), perceived ability to deal with the threat (perceived good health and a supportive network), and a desire to inform relatives (being a parent). The two approaches to educating patients, viz. direct patient education vs. education via their physician, did not significantly differ in terms of percentages of patients receiving counseling (42% vs. 51%, respectively) or the percentage choosing DNA testing (32% vs. 37%, respectively). PMID- 12537653 TI - Comprehensive genetic counseling for families at risk for HNPCC: impact on distress and perceptions. AB - The aim of the study was to explore distress and health beliefs before and after comprehensive interdisciplinary counseling in families at risk for hereditary non polyposis colorectal cancer (HNPCC). Results reported here were derived from a consecutive sample of 65 counselees [31 patients with colorectal cancer (CRC) and 34 unaffected at-risk persons] who participated in interdisciplinary counseling provided by human geneticists, surgeons, and psycho-oncologists before genetic testing. Data were collected from self-administered questionnaires before, as well as 4-6 weeks after, counseling. Distress and perceptions specific to HNPCC were assessed at both timepoints using standardized as well as author-derived instruments. Distress declined after counseling, as did worries related to HNPCC. An increase was found in personal belief in control of cancer risk, for instance, in the perceived efficacy of early detection of CRC. We also observed a trend toward greater anticipated ability to cope with a positive gene test after counseling. Changes after counseling were generally more pronounced for persons at risk, as compared to patients with cancer. The decrease in distress was partly attributable to an increase in personal self-confidence. One-third of the sample reported enhanced communication specific to hereditary disease within the family after counseling. A substantial minority, however, said they experienced increased worry and physical symptoms after counseling. Overall, counselees demonstrated less stress and perceived cancer threat as well as enhanced beliefs regarding personal control over cancer, suggesting an overall beneficial impact of comprehensive counseling. Further research is needed to identify those individuals most at risk for increased fear and worry related to HNPCC so that they may be most appropriately counseled. PMID- 12537654 TI - Impact of genetic counseling and testing on colorectal cancer screening behavior. AB - One goal of cancer genetic counseling is to improve early detection and prevention of cancers by identifying individuals at risk and providing screening recommendations. This study determined the impact of genetic counseling and testing on patient's post-genetic risk assessment colorectal cancer screening behaviors. Follow-up data from patients seen August, 1996, through May, 1998, at the Johns Hopkins Cancer Risk Assessment Clinic were analyzed. Eligible patients included those without cancer who were due for a colon examination by the time of follow-up, based on recommendations given during genetic risk assessment (GRA). We analyzed the role of gender, age, time since GRA, prior screening, genetic testing decision, mutation status, and post-GRA screening. Of 65 patients evaluated, 50 (76.9%) had undergone at least one endoscopic colon exam prior to visiting the Cancer Risk Assessment Clinic. At the time of GRA, 37 of 65 (56.9%) were overdue for a colon exam and at the time of follow-up, 15/65 (23.1%) were past due (p < 0.001). Patients with mutation-positive genetic tests were more likely to adhere to screening guidelines than those with negative gene tests (100% vs. 40.5%, p = 0.05). Genetic counseling and testing increases overall patient adherence with recommended colon screening, especially for those with positive genetic test results. However, patients with negative results may receive false reassurance about cancer risks and fail to follow recommended screening. Emphasis should be placed on the importance of screening even when genetic test results are negative. PMID- 12537655 TI - Likelihood of attending bowel screening after a negative genetic test result: the possible influence of health professionals. AB - This study was undertaken to determine the extent to which the reported likelihood of attending future bowel screening following negative genetic testing results for familial adenomatous polyposis (FAP) varies between the type of health professional providing care and the country of testing. The study subjects were 103 unaffected adults at risk for FAP who received negative results following predictive DNA testing. Our study indicates that the reported likelihood of attending bowel screening was higher in those given results by nongenetics physicians, rather than by genetics professionals; the reported likelihood of attending bowel screening under these circumstances was also higher in the UK than in Australia. Both of these results were affected by the perceived chances of developing FAP, and, in the case of the country of testing, by the perceived accuracy of the genetic test result and the perceived seriousness of the disease. How and what health professionals communicate with patients about genetic testing may explain the differences between type of health professional and country of testing and attitudes toward bowel screening. If this is the case, training in communication may change patients' perceptions and, in turn, their behavioral intentions and actions following a negative test result. PMID- 12537656 TI - Prevalence of the E1317Q variant of the APC gene in Italian patients with colorectal adenomas. AB - Loss of APC is an initial, rate-limiting event in inherited and sporadic colorectal tumorigenesis. Rare germline APC mutations have been identified in patients with multiple colorectal adenomas. Recently, the E1317Q APC variant has been associated with a predisposition to the development of multiple colorectal adenomas. In this study, the prevalence of the E1317Q variant was examined in 182 patients with single or multiple colorectal adenomas, and in 235 controls. In all, E1317Q was identified in two of 182 patients with adenomatous polyps (1.1%) and in two of 235 controls (0.8%) (p = 0.59). The risk of harboring adenoma(s) among subjects bearing the E1317Q variant was 1.29 (95% CI 0.09-18.0). No difference in the prevalence of E1317Q between cases with single (2.0%) or multiple colorectal adenomas (0.7%) and controls (0.8%) was found. None of the subjects with a family history of colorectal cancer carried the E1317Q variant. In conclusion, our results confirm that only a very small fraction of colorectal adenomas may be associated with the presence of E1317Q. PMID- 12537657 TI - An intronic polymorphism of the hMLH1 gene contributes toward incomplete genetic testing for HNPCC. AB - Hereditary non-polyposis colorectal cancer (HNPCC) is a common hereditary cancer. Genetic testing is complicated by the multiple DNA mismatch repair genes that underlie the disorder. Many suspected HNPCC families have no germ-line mutation identified. We reassessed an unusual family that appeared to have 2 individuals homozygous for a germline mutation within exon 1 of the hMLH1 gene. A few rare individuals with two inherited mutations in one of the mismatch repair genes have been reported and appear to have a distinct clinical appearance. However, there were no clinical features in the family discussed here that were consistent with constitutive lack of hMLH1. Redesigning the intronic primers for exon 1 identified a common polymorphism located within the original intronic primer site. The polymorphism prevented amplification of the wild-type allele, giving the erroneous appearance of homozygous inheritance of the mutated allele. Likewise, common intronic polymorphisms, if located within primer sequences on the chromosome harboring the HNPCC germ-line mutation could restrict amplification to only the wild-type allele, which may contribute significantly to the low success rate of identifying mutations in HNPCC families. PMID- 12537658 TI - Mutational analysis of the hMSH6 gene in familial and early-onset colorectal and endometrial cancer in Israeli patients. AB - Familial colorectal cancer (CRC) is noted in about 15% of incident CRC cases, and at times is hallmarked by an age at diagnosis less than 50 years. Familial adenomatous polyposis (FAP) and hereditary non-polyposis colon cancer (HNPCC) account for about 40% of familial cases. Thus, the majority of familial and early onset CRC remain genetically elusive. Similarly, the majority of familial and early onset endometrial cancer (EC), the most prevalent extracolonic tumor in HNPCC, are genetically undefined. An attractive candidate is the hMSH6 gene. Israeli patients with early onset (age under 50 years) (n = 44) and familial nonsyndromic (n = 23) CRC, and women with familial clustering of EC or CRC (n = 12), and those diagnosed with EC at, or under, the age of 50 years (n = 5) were genotyped for germ-line mutations within the hMSH6 gene. Exon-specific PCR was followed by denaturing gradient gel electrophoresis (DGGE) analysis, complemented by DNA sequencing of abnormally migrating fragments. No patients displayed a truncating mutation, and 1 CRC patient harbored a novel missense mutation (V878A). In addition, 6 previously described polymorphisms were detected. In conclusion, mutations in the hMSH6 gene occur uncommonly in Israeli patients with familial and early-onset CRC and EC. PMID- 12537659 TI - Frequency of hemochromatosis C282Y and H63D mutations in Sardinia. AB - Hereditary hemochromatosis (HH) is one of the most common autosomal recessive disorders of iron metabolism among Caucasians, and it is associated with C282Y mutation of the HFE gene in populations of Celtic origins. A second mutation, H63D, shows a very high widespread frequency, although its role in iron metabolism is still inconclusive. There are no data on the frequencies of these two mutations in Sardinia, an island in the Mediterranean sea that has not been invaded by Celtic peoples. We examined 836 chromosomes from Sardinian subjects and tested for the mutation by restriction enzyme digestion of PCR products. Among the 836 analyzed chromosomes, we found a C282Y allele frequency of 0.0036 and an H63D allele frequency of 0.173. These data could explain the observed rarity of HH in Sardinia. The high allele frequency of H63D and the rarity of HH in Sardinia is suggestive that this mutation is not a major contributor to this disease. PMID- 12537660 TI - A very rare association of three mutations of the HFE gene for hemochromatosis. AB - In the present paper, we describe a patient who is a compound heterozygote for three mutations in the HFE gene: C282Y, H63D, and E168Q. The patient's mother carries two copies of H63D and one copy of E168Q; the patient's father is heterozygous for C282Y. The family study indicates that the patient, as well as his sister, a maternal uncle, and a first cousin, all have inherited a single HFE allele that contains two mutations H63D and E168Q. The clinical symptoms and laboratory findings of the patient and his relatives are consistent with the conclusion that the E168Q mutation by itself is unlikely to result in hemochromatosis. PMID- 12537661 TI - Molecular screening of FRAXA and FRAXE in Indian patients with unexplained mental retardation. AB - Fragile-X mental retardation is the commonest form of inherited mental retardation. We have studied 146 Indian patients (174 X chromosomes) with unexplained mental retardation by molecular methods. All study subjects were unrelated. Three of the 118 males were found to have the FMR1 full mutation. None of the patients tested were positive for the FMR2 full mutation. The Fragile X prevalence was 2.5% among males, which is lower than previously reported in Indian mentally retarded patients. Screening for Fragile X among patients with nonspecific mental retardation is important, even if there is no family history of mental retardation or typical behavioral or physical features associated with the Fragile-X phenotype. Identification of positive cases is also very important for the families, because of the high recurrence risk of the disease. Large multicenter screening programs with uniform criteria would be worthwhile to determine the prevalence of Fragile-X mental retardation in the Indian population. PMID- 12537662 TI - Chromosome 6. PMID- 12537663 TI - Year 2002: the best year yet for CB&R. PMID- 12537664 TI - Advances in dendritic cell-based vaccine of cancer. AB - Dendritic cells (DCs) are potent antigen presenting cells that exist in virtually every tissue, and from which they capture antigens and migrate to secondary lymphoid organs where they activate naive T cells. Although DCs are normally present in extremely small numbers in the circulation, recent advances in DC biology have allowed the development of methods to generate large numbers of these cells in vitro. Because of their immunoregulatory capacity, vaccination with tumor antigen-presenting DCs has been proposed as a treatment modality for cancer. In animal models, vaccination with DCs pulsed with tumor peptides, lysates, or RNA or loaded with apoptotic/necrotic tumor cells could induce significant antitumor CTL responses and antitumor immunity. However, the results from early clinical trails pointed to a need for additional improvement of DC based vaccines before they could be considered as practical alternatives to the existing cancer treatment strategies. In this regard, subsequent studies have shown that DCs that express transgenes encoding tumor antigens are more potent primers of antitumor immunity both in vitro and in vivo than DCs simply pulsed with tumor peptides. Furthermore, DCs that have been engineered to express certain cytokines or chemokines can display a substantially improved maturation status, capacity to migrate to secondary lymphoid organs in vivo, and abilities to stimulate tumor-specific T cell responses and induce tumor immunity in vivo. In this review we also discuss a number of factors that are important considerations in designing DC vaccine strategies, including (i) the type and concentrations of tumor peptides used for pulsing DCs; (ii) the timing and intervals for DC vaccination/boostable data on DC vaccination portends bright prospects for this approach to tumor immune therapy, either alone or in conjunction with other therapies. PMID- 12537665 TI - Rituximab-mediated sensitization of B-non-Hodgkin's lymphoma (NHL) to cytotoxicity induced by paclitaxel, gemcitabine, and vinorelbine. AB - Non-Hodgkin's lymphoma (NHL) responds initially to chemotherapy, however, the disease often relapses and acquires a chemoresistant phenotype. Third line treatment options such as paclitaxel, gemcitabine, and vinorelbine have been tried in patients with refractory or relapsed NHL with antilymphoma activity. Currently, rituximab (anti-CD20) has been approved for treatment of indolent NHL, with significant activity in a wide spectrum of B-cell malignancies, though a percentage of patients do not respond to rituximab treatment. Previous findings from our laboratory have demonstrated that rituximab can sensitize drug-resistant NHL B-lymphoma in vitro to some chemotherapeutic drugs. Hence, in this study, we examined the effect of combination treatment of rituximab and the drugs paclitaxel, gemcitabine, and vinorelbine on various NHL cell lines. Our findings indicate that pretreatment of NHL tumor cells with rituximab sensitizes drug resistant NHL tumor cells to drug-mediated cytotoxicity. These findings suggest the potential clinical application of combination treatments of rituximab and paclitaxel, gemcitabine or vinorelbine in the treatment of drug/rituximab refractory non-Hodgkin's lymphoma. PMID- 12537666 TI - Expression of zeta in T cells prior to interleukin-2 therapy as a predictor of response and survival in patients with ovarian carcinoma. AB - Expression levels of T-cell receptor (TcR)-associated zeta chain were reported to reflect functional competence of T lymphocytes in patients with cancer. This retrospective study was performed to evaluate zeta chain expression in circulating T cells obtained from clinical responders and nonresponders among 19 patients with advanced ovarian carcinoma treated with intraperitoneal interleukin 2 (IL-2) biotherapy. Banked lymphocytes, which were collected from the patients who participated in a phase I clinical trial performed between 1987 and 1990, were used for quantitative flow cytometry to measure zeta-chain expression in T lymphocytes prior to and at the end of therapy. The data were correlated with 7 year survival. The patients (9 responders and 10 nonresponders) were stratified into two groups based on zeta chain expression in CD3+ T cells above or below the mean. Patients with lower zeta expression in circulating T cells had shorter survival compared to patients whose T cells expressed high zeta. Pretherapy zeta expression was significantly lower (p = 0.03) in CD8+T cells of nonresponders than in CD8+T cells of normal controls. In patients with advanced ovarian carcinoma, low expression of the zeta chain in peripheral blood T cells prior to biotherapy might both reflect a large tumor burden and predict a poor of response to IL-2 bio-therapy. PMID- 12537667 TI - Comparison of a monomeric and dimeric radiolabeled RGD-peptide for tumor targeting. AB - The alpha v beta 3 integrin, a transmembrane heterodimeric protein expressed on sprouting endothelial cells, binds to the arginine-glycine-aspartic acid (RGD) amino acid sequence of extracellular matrix proteins such as vitronectin. Growing malignant tumors continuously require angiogenesis. As a result, alpha v beta 3 is preferentially expressed in growing tumors and is a potential target for radiolabeled RGD-peptides. In this study we compared the tumor targeting characteristics of a monomeric radiolabeled RGD-peptide with those of a dimeric analogue. Both peptides were radiolabeled with 99mTc via the hydrazinoni cotinamid (= HYNIC) moiety to form 99mTc-HYNIC-c(RGDfK) and 99mTc-HYNIC-E [c(RGDfK)]2. In vitro, the IC50 showed a 10-fold higher affinity of the dimer for the alpha v beta 3 integrin as compared to the monomer (0.1 vs. 1.0 nM). In athymic female BALB/c mice with subcutaneously growing OVCAR-3 ovarian carcinoma xenografts, tumor uptake peaked at 5.8 +/- 0.7% ID/g and 5.2 +/- 0.6% ID/g for the dimer and the monomer, respectively. At 1, 2, and 4 h postinjection (p.i.) uptake of the dimer in the tumor was significantly higher than that of the monomeric analogue. Tumor-to-blood ratios were highest at 24 h p.i. at a value of 63 for both compounds. At all timepoints kidney retention of the dimer was significantly higher as compared to kidney retention of the monomer. In conclusion, in this mouse model the dimeric RGD-peptide showed better retention in the tumor than the monomeric analogue, most likely due to the bivalent interaction with the target cell. Furthermore, kidney retention of the dimeric peptide was higher than that of the monomeric peptide. PMID- 12537668 TI - BCL2 antisense reduces prostate cancer cell survival following irradiation. AB - Irradiation of the prostate, delivered as external beam radiation therapy (EBRT), is currently one of the few treatment options for localized prostate cancer. While it is relatively effective, the failure rate still remains unacceptably high with a 5-year biochemical failure rate of 10-40%. Utilizing genetically engineered LNCaP prostate cancer sublines that either overexpress Bcl2 (LNCaP/S22 d) or have down-regulated Bcl2 (LNCaP/AS17-f) we investigated the influence of this antiapoptotic protein on clonogenic survival following radiation. The radiation dose response curves (2-8 Gy) for the sublines differed significantly from the parental LNCaP (LNCaP/S22d: p < 0.001 and LNCaP/AS17-f: p = 0.008). The relative survival of the sublines revealed increased survival in the Bcl2 overexpressing cells, and decreased survival in the Bcl2 down-regulated cells. These data suggest a potentially important therapeutic approach for enhancing radiosensitivity in prostate tumors via antisense oligonucleotide or other drug therapies that down-regulate Bcl2. Strategies such as these likely hold the promise of enhancing the efficacy of EBRT by decreasing tumor cell survival, reducing the incidence of tumor recurrence and improving patient outcome. PMID- 12537669 TI - Application of a yeast assay to detect functional p53 mutations in archival prostate cancer tissue. AB - Detection and functional evaluation of mutant p53 alleles using a yeast assay could yield significant information for predicting the prognosis of patients with prostate cancer (CaP). Since the current version of this yeast assay is not applicable to archival tissues, we developed a modified assay for use on formalin fixed, paraffin-embedded tissue and have applied it to the study of patient samples. Using this modified assay, we examined archival CaP samples from 10 patients for mutations in exons 5-8 of p53 gene. Mutations were detected in four samples: three resulted in the formation of red yeast colonies indicating complete loss of function, while one gave pink yeast colonies, indicating that this mutant retained partial function. In parallel, we analyzed these samples for p53 abnormalities using a single-strand conformational polymorphism (SSCP) approach. Only three of the four yeast-positive samples gave abnormal SSCP bands. In each case where abnormal p53 was found by both methods, DNA sequencing revealed the identical base change. These results suggest that the modified yeast assay may be more sensitive than SSCP for detection of p53 mutations, and demonstrate that the modified method can be used to detect and evaluate the function of p53 mutants present in archival tissue. PMID- 12537671 TI - Active immunotherapy with anti-idiotypic antibody for patients with nasopharyngeal carcinoma (NPC). AB - Two anti-idiotypic monoclonal antibodies (Ab2), designated 2H4 and 5D3, against two antitumor antibodies Ab1 (FC2 and HNL5) that recognize nasopharyngeal carcinoma (NPC) associated antigen were generated. They could substitute NPC antigen to induce humoral and cellular immune response against NPC cells in syngeneic mice. Nineteen patients with NPC at stage IV were chosen for active immunotherapy. They were treated with aluminum hydroxide-precipitated Ab2 2H4 or 5D3 accompanying radiotherapy. None of the immunization of anti-idiotypic monoclonal antibody (mAb) was associated with toxicity or allergies reactions. Nine patients with radiotherapy alone served as control. Both anti-anti-idiotypic antibodies (Ab3) and anti-NPC antibodies (Ab1') were increased and human anti mouse Ig antibodies (HAMA) occurred in nineteen patients of the experimental group; whereas the levels of Ab1' did not rise in the control group. Serum IL-2, IFN-gamma, and TNF-alpha levels were increased in most patients in the experimental group, while in the control group, there were no differences of Ab1' and cytokine level between pretherapy and posttherapy. In addition, IL-2 mRNA expression in peripheral blood mononuclear cells (PBMC) of NPC patients was closely related to serum IL-2 (r = +0.8829) by in situ hybridization. Therefore, mouse anti-idiotypic antibodies 2H4 and 5D3 are safe for active immunotherapy and might enhance humoral and/or cellular immunity of NPC patients receiving radiotherapy. PMID- 12537670 TI - Fusion of the CH1 domain of IgG1 to epidermal growth factor (EGF) prolongs its retention in the blood but does not increase tumor uptake. AB - An expression vector (pJW4) for a human epidermal growth factor (hEGF)-CH1 fusion protein was constructed by fusing the gene for hEGF with the gene for CH1 of murine IgG1 with/without a peptide linker sequence [(GGGGS)3] and inserting the recombinant gene into vector pGEX2T. Expression vector pGEX2T was transfected into E. coli (BL-21) and hEGF-CH1 expressed by induction of the lac Iq promotor with 50 microM isopropyl beta-D-thiogalactopyranoside (IPTG). hEGF- CH1 fused to glutathione S-transferase (GST) was isolated and purified by affinity chromatography. GST was cleaved using thrombin. SDS-PAGE demonstrated a protein with the expected M(r) (18 kDa) positive for hEGF by Western blot. hEGF-linker CH1 exhibited preserved binding to A431 (2-3 x 10(6) EGFR/cell) and MDA-MB-468 breast cancer cells (1-2 x 10(6) EGFR/cell). hEGF-CH1 without the linker exhibited poor receptor binding. hEGF-linker-CH1 also exhibited strong binding to soluble EGFR equivalent to that of hEGF. The tumor and normal tissue distribution of hEGF-linker-CH1 labeled with 123I was compared with 123 I-hEGF at 24 h after i.v. injection to mice implanted with s.c. MDA-MB-468 xenografts. Fusion of hEGF with CH1 increased its retention in the blood 14-fold but did not significantly increase tumor uptake. Tumor/blood ratios were higher for hEGF than for hEGF linker-CH1. We conclude that hEGF is more attractive than hEGF-linker-CH1 for imaging EGFR-positive tumors. PMID- 12537672 TI - Radioimmunotherapy with 186Re-labeled monoclonal antibody to treat liver metastases of colon cancer cells in nude mice. AB - The efficacy of radioimmunotherapy (RIT) in the treatment of minimal disease has been previously shown, despite the limitation of beta-emitters suggested by a mathematical model. In the present study, the efficacy of RIT with an anti colorectal cancer IgG1 A7 conjugated with 186Re was examined in a liver metastasis model established by intrasplenic inoculation of human colon cancer cells. In this model, small metastases of less than 1 mm in diameter can be observed 1 week after cell inoculation. Metastases attain a diameter of several millimeters at 2 weeks. 186Re-A7 accumulated exclusively in metastases, displaying a value of 24.1 +/- 8.7% ID/g 2 days after the injection. 186Re-A7 accumulation in liver metastases increased with decreasing size. RIT with 7 MBq 186Re-A7 at 2 weeks significantly suppressed the growth of metastases; weight of metastases 4 weeks after cell inoculation was 5.96 +/- 0.87 g in nontreated control mice and 1.25 +/- 0.75 g in mice receiving 186Re-A7 RIT (p < 0.0001). RIT at 1 week more effectively inhibited metastatic growth to 0.08 +/- 0.05 g (p < 0.002 vs. RIT at 2 weeks). RIT with a class-matched irrelevant MAb 186Re-HPMS-1 at 1 week after cell inoculation somewhat suppressed metastatic growth, 3.39 +/- 0.25 g at 4 weeks, as compared with the control; however, 186Re-HPMS-1 RIT was far less effective than 186Re-A7 RIT (p < 0.0001). These results support the use of RIT with 186Re-MAb in an adjuvant setting in cases involving minimal disease. Factors such as higher and homogeneous MAb accumulation in small nodules, better perfusion, and subsequent better oxygenation likely compensate for the loss of beta radiation outside small metastases. PMID- 12537673 TI - Radioimmunoscintigraphy (RIS) with bectumomab (Tc99m labeled IMMU-LL2, Lymphoscan) in the assessment of recurrent non-Hodgkin's lymphoma (NHL). AB - The efficacy of a Tc99m-labeled anti-lymphoma antibody fragment, bectumomab [LymphoScan], was retrospectively examined in the staging of recurrent or newly diagnosed non-Hodgkin's lymphoma (NHL) [7 patients] and to assess targeting before radioimmunotherapy (RIT) [14 patients]. Performance was graded relative to conventional imaging. Tumors included 7 low-grade, 11 intermediate-grade, and 3 high-grade histologic subtypes. Computed x-ray tomography, radiogallium imaging, FDG-PET, and bone marrow biopsy defined 117 sites. Bectumomab revealed 56% of these sites. In 4 patients bectumomab uncovered five sites not evident by conventional imaging. In addition, it uncovered one site in the brain, an area not covered in the standard work-up of asymptomatic patients. Bectumomab imaging most often failed in central abdominal and thoracic locations, and excelled in revealing disease in the head and neck. Relative to Ga67 citrate imaging, the performance of bectumomab was variable, with no clear relation to anatomic location; there was better targeting of low and intermediate grade NHL. Radiogallium out-performed bectumomab imaging in 23 sites, 19 of which were in patients with high or intermediate-grade disease. Bectumomab was superior to radiogallium at six sites, five of which involved low-grade tumor. CONCLUSION: Bectumomab shows promise as a pre-RIT probe for targeting of B-cell NHL. It excelled at defining small volume, low-grade disease. However, as a purely diagnostic agent, its performance was variable. PMID- 12537674 TI - Quantitative radioimmunoimaging for radioimmunotherapy treatment planning: effect of reduction in data sampling on dosimetric estimates. AB - Quantitative radioimmunoimaging (serial anterior/posterior imaging and blood sampling) is useful for radioimmunotherapy treatment planning, but can be quite time consuming. To predict whether accurate radiation absorbed dose estimates can be maintained with a reduction in data sampling, 12 patients undergoing indium 111/yttrium-90 anti-CD20 monoclonal therapy for whom absorbed doses were estimated based on eight data samples (acquired at 0, 2, 4, 24, 48, 72, 96, and 144 h, respectively), were retrospectively reanalyzed using only five samples (0, 4, 24, 72, and 144 h, respectively). Calculated residence times (in h) and absorbed doses (in cGy), for the whole body, kidneys, liver, lungs, spleen, and red marrow were compared with the original values based on the eight samples using Student's paired t-test. Linear regression and Bland-Altman analysis of the two data sample groups was also performed. The mean residence times in the five- and eight-data samples groups were essentially the same (17.7 +/- 26.6 h [range, 0.3-79.0 h] versus 17.6 +/- 26.6 h [range, 0.3-79.5 h]; p = 0.72), as were the mean absorbed doses (336 +/- 411 cGy [range, 38-2434 cGy] versus 325 +/- 381 cGy [range, 39-2246 cGy]; p = 0.24). Also, the linear regressions were excellent (residence time y = 1.00x + 0.09 h [r = 0.99]; absorbed dose y = 1.06x - 7.74 cGy [r = 0.98]). Additionally, Bland-Altman analysis revealed no significant sample bias in residence time (0.03 +/- 0.68 h, 0.9% +/- 10.0) or absorbed dose (11 +/- 76 cGy, 1.0% +/- 9.3). These results demonstrate that reduced data sampling in quantitative radioimmunoimaging can be achieved without significantly altering radiation absorbed dose estimates, but with a significant savings in imaging, blood sampling, and processing time. PMID- 12537675 TI - Intraocular pressure fluctuations in medical versus surgically treated glaucomatous patients. AB - The purpose of this study was to compare the IOP fluctuations in the daytime tension curves of glaucoma patients under ocular hypotensive therapy with those of patients previously submitted to trabeculectomy. We also evaluated the IOP peaks and fluctuations for the same patients in response to the water-drinking test (WDT). The study included 30 primary open-angle glaucoma (POAG) patients using ocular hypotensive medications and with no history of previous intraocular surgery (medical group), and 30 POAG patients previously submitted to one or more trabeculectomies though taking no medication at the time of the study (surgical group). All patients were submitted to a diurnal tension curve--DTC (8:30-17:00/3 hour intervals) followed by the WDT. The IOP peak and IOP fluctuation during the diurnal tension curve were significantly greater in the medical group than in the surgical group. The same was observed following the WDT. From an overall baseline IOP of 10.6 mmHg, the mean IOP change following the WDT was 13% in the surgical group and 40% in the medical group. Patients submitted to trabeculectomy have less TOP fluctuations during the diurnal tension curve and following a water drinking provocative test. This effect could represent an additional benefit of surgery in controlling the intraocular pressure of glaucomatous patients. PMID- 12537676 TI - A prospective, long-term, randomized study of the efficacy and safety of the drug combination pilocarpine 1% with clonidine 0.06% or clonidine 0.125% versus timolol 0.25%. AB - The purpose of this prospective, randomized study was to evaluate the efficacy and safety of the drug combinations--pilocarpine 1% with clonidine 0.06% and pilocarpine 1% with clonidine 0.125%, in comparison with timolol 0.25%. 54 patients with bilateral primary open angle glaucoma and comparable IOP were assigned to three study groups: group 1 received the drug combination of pilocarpine 1% and clonidine 0.06%, group 2 received the drug combination pilocarpine 1% and clonidine 0.125% and group 3 only timolol 0.25% drops. In all groups twice-daily medication was used. Mean diurnal IOP as well as ocular and systemic side effects were evaluated at week 1, 2, 4, 8 and 12. Changes in blood pressure, pulse rate, pupil size and patient symptoms were also recorded. At three months all patients had completed the study. Diurnal IOP was significantly reduced from baseline in all groups and consistent IOP reduction was achieved in all three groups at all follow up periods. Mean IOP reduction was 18.40%, 28.45% and 24.64% in group 1, group 2 and group 3, respectively. Flattening of the diurnal fluctuation in IOP was also seen in all patients. On comparing the IOP reduction achieved amongst the three groups, there was a statistically significant difference between group 1 and group 2 as well as between group I and group 3 while the difference between group 2 and group 3 was statistically insignificant. The drug combinations of pilocarpine 1% and clonidine did not produce any statistically significant effect on pupil diameter. No statistically significant difference was noted with respect to patient symptoms, blood pressure and pulse rate. In conclusion, the drug combination of pilocarpine 1% and clonidine 0.125% produces IOP reduction comparable to that achieved with timolol 0.25% drops in twice daily dosage and does not result in any significant ocular and systemic adverse effects. PMID- 12537677 TI - Biodegradable calcium phosphate nanoparticles as a new vehicle for delivery of a potential ocular hypotensive agent. AB - The purpose of this study was to determine the efficacy of a newly prepared formulation containing biodegradable calcium phosphate nanoparticles (CAP) and 7 hydroxy-2-dipropyl-aminotetralin (7-OH-DPAT) in pigmented and non-pigmented rabbits using the surrogate end points of intraocular pressure (IOP) and aqueous flow rate. IOP (mmHg) was measured by utilizing a manometrically calibrated Mentor pneumatonometer. Rates of aqueous humor flow were measured with a Fluorotron Master by estimating the dilution rate of fluorescein. In non pigmented rabbits, the ocular hypotension induced by topical administration of 7 OH-DPAT (75 microg) with CAP (115 microg) was more pronounced and sustained than that of 7-OH-DPAT without CAP. Furthermore, IOP-lowering effects of topically administered 7-OH-DPAT (125 microg) alone were markedly diminished in pigmented rabbits compared to non-pigmented rabbits. However, topical application of 7-OH DPAT formulated with CAP produced significant dose-related (37.5, 75, 125 microg) reductions of IOP accompanied by suppression of aqueous humor flow rates in pigmented rabbits. It is postulated that 7-OH-DPAT in vehicle without CAP binds to pigments in the anterior segment of the pigmented rabbit's eyes, and this binding limits the 7-OH-DPAT's action. Pretreatment with raclopride, a dopamine D2/D3 receptor antagonist, reduced the ocular hypotensive effect induced by 7-OH DPAT in vehicle containing CAP thereby supporting the role for dopamine D2/D3 receptors in modulating IOP. It is concluded that CAP, as a delivery system, enhances activity by 7-OH-DPAT in pigmented rabbit eyes suggesting that CAP is potentially useful for achieving controlled and targeted drug delivery for treatment of ocular diseases. PMID- 12537678 TI - Effects of kallidinogenase on ocular tissue circulation in rabbits. AB - The purpose of this study is to evaluate the effects of kallidinogenase, a tissue kallikrein, on tissue circulation in the optic nerve head (ONH), choroid and retina. Kallidinogenase (1.0 IU/kg) or saline was injected intravenously into urethane-anesthetized rabbits, and the normalized blur value (NB), a quantitative index of in vivo tissue blood velocity, was measured in the ONH, choroid and retina before and for 90 minutes after injection, using the laser speckle method. The difference in NB values in the kallidinogenase group was significantly higher compared with that in the control group in the choroid (p < 0.05) and retina (p < 0.05, ANOVA of repeated measurements). In the ONH, however, there was no significant difference between the kallidinogenase group and the control group except transient increment 10 min after drug administration (p < 0.01, unpaired t test). On the other hand, systemic condition parameters and intraocular pressure showed no intergroup difference significantly except for a transient decrease in blood pressure and increase in pulse rate in the kallidinogenase groups. Kallidinogenase increased blood velocity, and probably blood flow, in the choroid and retina of rabbits. PMID- 12537679 TI - Effect of different dose schedules of bimatoprost on intraocular pressure and pupil size in the glaucomatous Beagle. AB - The changes in intraocular pressure and pupil size in glaucomatous dogs were evaluated after instillations of 0.03% bimatoprost (Lumigan, Allergan, Irvine, CA USA) once in the morning, or once in the evening, or twice daily in five day multiple dose studies. Applanation tonometry (IOP) and pupil size (PS) measurements were obtained at 8 am, 10 am, 12 noon, 2 pm, and 4 pm in 8 glaucoma dogs. Methylcellulose (0.5% as placebo) was instilled in the control eye, and 0.03% bimatoprost was instilled in the opposite drug eye. Methylcellulose (0.5%) and 0.03% bimatoprost were instilled the second through the fifth days with instillations in the morning (8:30 am), or evening (8 pm), or twice daily (8:30 am and 8 pm). The mean +/- SEM diurnal changes in IOP from baseline values after 0.03% bimatoprost at 8 am once daily for the next four days were 25.0 +/- 3.2 mm Hg, 25.6 +/- 2.9 mm Hg, 25.5 +/- 3.0 mm Hg, and 26.0 +/- 3.2 mm Hg respectively, and were significantly different from the control eye. After bimatoprost was instilled at 8 pm, the mean +/- SEM changes in IOP from baseline values in the drug eyes were 27.3 +/- 2.4 mm Hg, 26.6 +/- 2.2 mm Hg, 27.2 +/- 2.5 mm Hg, and 27.3 +/- 2.6 mm Hg respectively. When 0.03% bimatoprost was instilled twice daily, the mean +/- SEM changes in IOP from baseline values were 39.1 +/- 2.3 mm Hg, 39.9 +/- 2.2 mm Hg, 39.9 +/- 2.3 mm Hg, and 39.6 +/- 2.1 mm Hg respectively, and were significantly different from the control eyes. Miosis of varying duration was frequent during the three studies. Bimatoprost instilled once daily (am or pm) as well as twice daily produces significant decreases in IOP and PS in the glaucomatous Beagle. PMID- 12537680 TI - Corneal permeation of ganciclovir: mechanism of ganciclovir permeation enhancement by acyl ester prodrug design. AB - Ganciclovir (GCV), a promising antiviral compound, has poor ocular bioavailability as a result of its relatively low partition coefficient. In this study, lipophilic ester prodrugs of GCV were synthesized in an effort to improve its uptake into ocular tissues. In vitro permeability studies were conducted on isolated rabbit corneal membranes using aliphatic mono-acyl ester prodrugs of GCV to determine the effect of lipophilicity and corneal hydrolysis rate on transcorneal diffusion. The GCV prodrugs showed a progressive decrease in solubility and a corresponding increase in Log P values as the chain length was ascended. Permeation studies using freshly isolated rabbit corneas showed that all prodrugs permeated as intact prodrug as well as hydrolyzed GCV. Corneal permeability coefficients increased with increasing lipophilicity for mono-ester prodrugs having more than three carbon atoms in the side chain. The permeability of GCV increased about 6-fold in ascending from the parent drug-ganciclovir (3.82 +/- 0.19 x 10(-6) cm sec(-1)) to its valerate ester prodrug (23.70 +/- 1.36 x 10( 6) cm sec(-1)). Among the prodrugs studied, the valerate ester showed the highest permeability and holds the most potential for development. Overall prodrug permeability correlated linearly with increased susceptibility of the GCV esters to undergo hydrolysis in the cornea. The present work indicates that the ideal prodrug is one that not only possesses enhanced partitioning characteristics, but also high enzyme susceptibility. Concentration of active GCV penetrating the corneal epithelium was substantially increased through the bio-reversible ester prodrug strategy. PMID- 12537681 TI - In vivo transscleral iontophoresis of amikacin to rabbit eyes. AB - The objectives of these studies were to determine the amount and distribution of the aminoglycoside antibiotic amikacin delivered to rabbit eyes following transscleral iontophoresis and to determine the inter-study reproducibility of delivery over three identical studies. New Zealand White rabbits (N = 6 per dose group) were treated with a 200-mg/mL amikacin solution at 0, 2, 3 or 4 mA of (+) DC current for 20 minutes. Amikacin concentrations in eye tissues were highest with the 4-mA treatment. Concentrations for all three studies at this current were approximately 5.4, 40, 41, 343, and 92 mcg/g in the vitreous humor, anterior segment, non-treated hemisphere of the sclera, treated hemisphere of the sclera, and retina/choroid, respectively. These values were approximately 27, 50, 40, 10, and 13 fold greater than in the 0-mA control group and are well above the in vitro minimum inhibitory concentrations (MICs) for this drug. Inter-study reproducibility (measured as %CV) depended on the tissue type and treatment group and ranged from 8% for the retina/choroid to 51% for the anterior segment in the 4-mA group. Pretreatment with topical proparacaine hydrochloride local anesthetic did not affect amikacin delivery and total drug delivered was not affected by delivery time for the same total charge administered. Therapeutically relevant amounts of amikacin were delivered into eye tissues in a reproducible and controllable manner. PMID- 12537682 TI - In vitro human scleral permeability of fluorescein, dexamethasone-fluorescein, methotrexate-fluorescein and rhodamine 6G and the use of a coated coil as a new drug delivery system. AB - PURPOSE: To determine the in vitro human scleral permeability of several dyes and drugdye combinations with varying molecular weights (MW) and lipid solubilities (fluorescein, dexamethasone-fluorescein, methotrexate-fluorescein, and rhodamine). Coils coated with rhodamine were also evaluated for scleral permeability and sustained release. METHODS: Scleral sections excised from moist chamber stored human globes were mounted in a 2-compartment perfusion chamber. A small depot of drug/dye (100 microl of 10(-4) M fluorescein, dexamethasone fluorescein, methotrexate-fluorescein or rhodamine) or a coated coil in 100 microl of BSS was added to the episcleral surface while perfusing BSS to the choroidal side. The perfusate was collected and measured for fluorescence. Permeability was calculated as Ktrans from the flux measurements. RESULTS: Ktrans values (cm/sec, mean +/- SE) for the studied dyes and drug-dye combinations were 5.21 +/- 0.71 x 10(-6) for fluorescein, 1.64 +/- 0.17 x 10(-6) for dexamethasone fluorescein, 3.36 +/- 0.62 x 10(-6) for methotrexate-fluorescein, 1.86 +/- 0.39 x 10(-6) for rhodamine and 2.18 +/- 0.23 x 10(-6) for the rhodamine from the coils. We found a significant difference between the permeability of the sclera to fluorescein and dexamethasone-fluorescein (P < 0.001), methotrexate-fluorescein (P < 0.05) and rhodamine (P < 0.001). Steady state flux was observed from the rhodamine coil. CONCLUSION: The rank order of scleral permeability to the studied dyes is as follows: fluorescein > methotrexate-fluorescein > rhodamine coil > rhodamine 6G > dexamethasone-fluorescein. Differences in scleral permeability are related to MW and lipid solubility. Prolonged transscleral diffusion of rhodamine delivered by solution and by coil are similar. PMID- 12537683 TI - Interactions of chlorpromazine with alpha-, beta- and gamma-crystallins. AB - The binding parameters (binding affinity constant, K and number of binding sites, p) has been determined spectrofluorometrically for chlorpromazine (CPZ) binding to the lens proteins--alphaL-crystallin, betaL-crystallin and gamma-crystallin. The binding affinity constants for CPZ binding to alphaL- and gamma-crystallins are higher than the binding affinity constants for 3betaL-crystallin, although the number of CPZ binding sites for betaL-crystallin is comparatively higher than the number for the other two lens proteins. CPZ causes local conformational changes around the tryptophan moieties of the protein molecules but does not cause any gross conformational change within the protein moieties. Binding of CPZ to alphaL-crystallin does not significantly alter the anti-aggregation properties of the molecular chaperone, alphaL-crystallin against oxidation-induced aggregation of gamma-crystallin at 37 degrees C and thermal aggregation of alcohol dehydrogenase (ADH) at 48 degrees C. Therefore, CPZ induced alteration in chaperone activity of alphaL-crystallin is probably not associated with the formation of cataracts. PMID- 12537684 TI - Creating high-level components with a generative representation for body-brain evolution. AB - One of the main limitations of scalability in body-brain evolution systems is the representation chosen for encoding creatures. This paper defines a class of representations called generative representations, which are identified by their ability to reuse elements of the genotype in the translation to the phenotype. This paper presents an example of a generative representation for the concurrent evolution of the morphology and neural controller of simulated robots, and also introduces GENRE, an evolutionary system for evolving designs using this representation. Applying GENRE to the task of evolving robots for locomotion and comparing it against a non-generative (direct) representation shows that the generative representation system rapidly produces robots with significantly greater fitness. Analyzing these results shows that the generative representation system achieves better performance by capturing useful bias from the design space and by allowing viable large scale mutations in the phenotype. Generative representations thereby enable the encapsulation, coordination, and reuse of assemblies of parts. PMID- 12537685 TI - Predicting nearest agent distances in artificial worlds. AB - In a number of multi-agent artificial life studies where agents interact over limited distances, the emergence and/or evolution of a specific behavior may depend critically upon interagent distances. Little theoretical analysis has been done previously concerning how to predict such distances. In this paper, we derive a probabilistic method that, for an agent at an arbitrary location in a two-dimensional cellular world, predicts the expected distance to a nearest other agent. Our method works for many world topologies, and we apply it to determine the expected distance for six commonly used ones. Further, the method is readily adapted to handle special restrictions. Over a wide variety of agent densities we show that the theoretically predicted distances are largely in agreement with the distances measured in computational experiments with randomly placed agents. We then utilize our prediction method to interpret recent observations that an imprecise threshold in the density of agents exists for the evolution of communication. We thus illustrate that, despite its conceptual simplicity, our method can aid the analysis and even the design of complex artificial environments populated by agents that have the potential to interact with one another. PMID- 12537686 TI - Genetic redundancy in evolving populations of simulated robots. AB - A number of authors have argued that redundancy in biological organisms contributes to their evolvability. We investigate this hypothesis via the experimental manipulation of genetic redundancy in evolving populations of simulated robots controlled by artificial neural networks. A genetic algorithm is used to simulate the evolution of robots with the ability to perform a previously studied task. Redundancy is measured using systematic lesioning. In our experiments, populations of robots with larger genotypes achieve systematically higher fitness than populations whose genotypes are smaller. It is shown that, in principle, robots with smaller genotypes have enough computational power to achieve optimal fitness. Populations with larger (redundant) genotypes appear, however, to be more evolvable and display significantly higher diversity. It is argued that this enhanced evolvability is a direct effect of genetic redundancy, which allows populations of redundant robots to explore neutral networks spanning large areas of genotype space. We conjecture that, where cost considerations allow, redundancy in functional or potentially functional components of the genome may make a valuable contribution to evolution in artificial and perhaps in biological systems. The methods described in the article provide a practical way of testing this hypothesis for the artificial case. PMID- 12537687 TI - "Artificial societies" and the social sciences. PMID- 12537690 TI - The role of the liver in the response to LPS: experimental and clinical findings. AB - The liver plays an important physiological role in lipopolysaccharide (LPS) detoxification and, in particular, hepatocytes are involved in the clearance of endotoxin of intestinal derivation. In experimental shock models, tumor necrosis factor (TNF)-alpha induces hepatocyte apoptosis and lethal effects are due to secreted TNF-alpha and not to cell-associated TNF-alpha. An exaggerated production of TNF-alpha has been reported in murine viral infections, in which mice become sensitized to low amounts of LPS and both interferon (IFN)-gamma and IFN-alpha/beta are involved in the macrophage-induced release of TNF-alpha. The prominent role of LPS and TNF-alpha in liver injury is also supported by studies of ethanol-induced hepatic damage. In humans, evidence of LPS-induced hepatic injury has been reported in cirrhosis, autoimmune hepatitis, and primary biliary cirrhosis and a decreased phagocytic activity of the reticulo-endothelial system has been found in these diseases. The origin of endotoxemia in hepatitis C virus (HCV) infected patients seems to be multifactorial and LPS may be of exogenous or endogenous derivation. In endotoxemic HCV-positive patients responsive to a combined treatment with IFN-alpha/ribavirin (RIB), endotoxemia was no longer detected at the end of the therapeutic regimen. By contrast, 48% of the non responders to this treatment were still endotoxemic and their monocytes displayed higher intracellular TNF-alpha and interleukin (IL)-1beta levels than responders. Moreover, in responders, an equilibrium between IFN-gamma and IL-10 serum levels was attained. In the non-responders, serum levels of IL-10 did not increase following treatment. This may imply that an imbalance between T helper (Th)1 and Th2 derived cytokines could be envisaged in the non-responders. PMID- 12537691 TI - LPS in microbial pathogenesis: promise and fulfilment. AB - Since LPS was discovered, all inquiry in the field was motivated by two articles of faith. First, it was held that LPS was an important factor in microbial pathogenesis. Second, it was held that the responses elicited by numerous molecules of microbial origin might be formally similar to the responses elicited by LPS. The identification of TLR4 as the core transducer of LPS responses, and the added discovery that other TLR paralogs sense other microbial products, has strongly validated these cherished beliefs. Moreover, it has amalgamated many separate lines of inquiry, and focused attention on the how the innate immune system perceives infection. PMID- 12537692 TI - Enhancement of endotoxin activity by muramyldipeptide. AB - Synthetic muramyldipeptide (MDP), the minimum structural moiety of bacterial peptidoglycan for adjuvant and related activities, sensitized mice for two types of lethal shock induced by lipopolysaccharide (LPS): an early anaphylactoid shock and late endotoxin shock. In relation to the late reaction in MDP-primed mice, enhanced production of inflammatory cytokines was induced in response to various bacterial components. MDP showed a priming effect in mice not only when administered parentally but also via the oral route. MDP activated human monocytic THP-1 cells in a CD14-, Toll-like receptor 2 (TLR2)- and TLR4 independent manner to increase expression of MyD88, a common adaptor and signaling molecule for TLRs, and exhibited synergistic cytokine inducing effects with TLR4 agonists (LPS, synthetic lipid A), TLR2 agonist (synthetic lipopeptide), and TLR9 agonist (bacterial CpGDNA) in THP-1 cells in culture. Consistent with these findings, MDP primed TLR2 knockout mice as well as wild type controls, but not TLR4-mutated C3H/HeJ mice, to enhance production of tumor necrosis factor-alpha upon stimulation with synthetic lipid A. In contrast to the BCG- and Propionibacterium acnes-priming system, MDP primed mice in an interferon gamma-independent manner. Further studies are required to elucidate the mechanisms of the synthetic and priming activities of MDP for various bacterial components. PMID- 12537693 TI - Structural and biological characterization of highly purified hepta-acyl lipid A present in the lipopolysaccharide of the Salmonella enterica sv. Minnesota Re deep rough mutant strain R595. AB - One major component of the Salmonella enterica sv. Minnesota Re deep rough mutant (strain R595) lipopolysaccharide is hepta-acyl lipid A (LA(hepta)). In a recent publication [Tanamoto K-I, Azumi S. Salmonella-type heptaacylated lipid A is inactive and acts as an antagonist of lipopolysaccharide action on human line cells. J Immunol 2000; 164: 3149-3156] the corresponding synthetic hepta-acyl lipid A (compound 516) was reported to be agonistically inactive but to rather suppress pro-inflammatory activation by the endotoxic hexa-acyl lipid A (LA(hexa), compound 506) and S-form LPS from Escherichia coli in the human macrophage-like cell lines THP-1 and U937. These results, however, were in contrast to previous findings with human mononuclear cells (hMNC) isolated from peripheral blood, in which compound 516 was found to be an agonist, expressing low, but significant, cytokine-inducing activity as compared to LA(hexa). We have investigated the structure of natural LA(hepta) from the S. enterica sv. Minnesota Re deep rough mutant strain (R595) by TLC immunoblot, MALDI-TOF mass spectrometry and NMR spectroscopy. Using these techniques, the structural identity between LA(hepta) and the synthetic compound 516 was confirmed. In corroboration of previous findings with studies employing compound 516, purified LA(hepta) was found to induce the production of TNF-alpha, IL-1beta and IL-6 in hMNC, thus displaying moderate agonistic activity. Furthermore, we showed that LA(hepta) agonistically activated nuclear translocation of NF-kappaB in THP-1 cells, thus clearly ruling out the possibility that LA(hepta) is an antagonist and that its biological activity is influenced by the type of human myeloid cells used for testing endotoxicity (hMNC versus THP-1 cells). PMID- 12537694 TI - Monoclonal antibodies against 3-deoxy-alpha-D-manno-oct-2-ulosonic acid (Kdo) and D-glycero-alpha-D-talo-oct-2-ulosonic acid (Ko). AB - Monoclonal antibodies (mAbs) were obtained after immunization of mice with neoglycoconjugates containing as a ligand the disaccharide Kdo(2-->4)Ko or Ko(2- >4)Kdo, representing structural elements of the core region of lipopolysaccharides (LPSs) from Acinetobacter haemolyticus and Burkholderia cepacia, respectively. One antibody, S67-9, bound with high specificity to Ko(2- >4)Kdo-BSA showing no cross reactivity with Kdo(2-->4)Kdo-BSA and the other antigens tested. A second mAb, S68-12, bound preferentially to Kdo(2-->4)Ko-BSA but cross reacted with Ko(2-->4)Kdo-BSA and Kdo(2-->4)Kdo-BSA. A third mAb, S67 27, was found to bind Kdo monosaccharide. Although mAbs S67-9 and S68-12 did not bind to LPS of Burkholderia or Acinetobacter as expected, the mAbs will be useful tools in studying the biosynthesis of LPS containing Ko. PMID- 12537695 TI - Requirement of TNF and TNF receptor type 2 for LPS-induced protection from lethal septic peritonitis. AB - Pretreatment of mice with low quantities of LPS induces endotoxin tolerance characterized by enhanced resistance to lethal doses of LPS and to a number of infectious challenges. Mice subjected to cecal ligation and puncture (CLP) survived the ensuing septic peritonitis significantly better when they had been pretreated with LPS. This LPS-induced protection was dependent on endogenous TNF production capacity since LPS pretreatment did not protect TNF-deficient mice from death after CLP. While mice deficient in the TNF receptor type 2 (p75TNFR) were as sensitive to CLP-induced mortality as control mice, LPS pretreatment could not reduce mortality in p75TNFR-deficient mice after CLP. Therefore, activation of the TNF receptor type 2 by endogenous TNF constitutes an important interaction for the development of LPS-induced resistance to bacterial infection. PMID- 12537696 TI - Lipopolysaccharide-cell interaction and induced cellular activation in whole blood of septic patients. AB - We used biotinylated LPS (LPSb) and flow cytometry to study LPS-monocyte interaction and LPS-induced cellular activation in whole blood from septic patients (SP). Expression of surface activation markers was evaluated on monocytes (HLA-DR) and T lymphocytes (CD69 and CD95), and intracellular TNF-alpha on monocytes. Saturating curve and kinetics of LPSb detection on monocytes were similar in SP and healthy volunteers (HV). LPSb bound to monocytes was detected after 5 min of incubation in both groups, with a more pronounced decay in SP. Monocytes from SP had a lower expression of HLA-DR as compared to HV, both constitutive and upon LPS stimulation. The proportion of monocytes producing TNF alpha after LPS stimulus was higher in HV than SP (mean +/- SD = 25.2 +/- 14.2% and 2.2 +/- 2.6%, respectively, P < 0.001). LPS-induced CD69 on T CD8+ and CD8- lymphocytes was similar for patients and controls. Expression of CD95 on T lymphocytes was higher in SP as compared to HV on T CD8+ cells (GMFI, mean +/- SD = 22.3 +/- 14.6 and 8.6 +/- 5.0, respectively, P = 0.01) and CD8- cells (GMFI, mean +/- SD = 28.3 +/- 7.7 and 14 +/- 4.3 respectively, P < 0.001). Thus, monocytes and lymphocytes seem to respond differently to LPS in septic patients. Monocyte hyporesponsiveness appears not to be related to a decreased binding capacity of LPS, but rather to an impaired signal transduction. PMID- 12537699 TI - Are antidepressants addictive? PMID- 12537698 TI - Does high mobility group 1 protein function as a late mediator for LPS- or TNF induced shock in galactosamine-sensitized mice? AB - The role of high mobility group-1 protein (HMG-1) in LPS- and TNF-alpha-induced lethal shock in galactosamine (GalN)-sensitized mice was investigated. No detectable HMG-1 levels were observed by immunoblotting analysis in plasma from untreated or GalN-sensitized BALB/c mice 5 h after LPS injection, although significant levels of HMG-1 were detected in plasma 6 h after the challenge. All GalN-sensitized BALB/c but not BALB/lps(d) mice succumbed by 6 h after LPS injection. When GalN-sensitized mice were injected with TNF-alpha, the presence of HMG-1 was seen at 5.5 h in plasma of BALB/c mice and at 6 h in BALB/lps(d) mice, although almost all GalN-sensitized BALB/c mice died by 6 h after challenge. The time-dependent phenomenon correlated with elevated serum aspartate aminotransferase (AST) levels and the appearance of apoptotic cells in livers. Administration of pooled plasma, equivalent to approximately 200 microg recombinant murine HMG-1, taken from mice on the verge of near death, did not result in induction of lethal shock in GalN-sensitized mice. Taken together with the late appearance of HMG-1 in moribund mice, these data suggest that HMG-1 does not decisively contribute to lethality in the GalN sensitization model. PMID- 12537697 TI - The role of the interferon regulatory factors, IRF-1 and IRF-2, in LPS-induced cyclooxygenase-2 (COX-2) expression in vivo and in vitro. AB - Cyclooxygenase (COX) exists as two isoforms: COX-1, which is constitutively expressed in most cell types; and COX-2, which is inducible by lipopolysaccharide (LPS) and cytokines in a variety of cell types. Although previous studies have implicated two DNA binding proteins, interferon regulatory factor (IRF)-1 and IRF 2, in the regulation of LPS- and IFN-gamma-induced COX-2, their effects in vivo and in vitro are not well-defined. Using real-time PCR, COX-2 gene expression in the livers and lungs of mice challenged in vivo and in macrophages stimulated with LPS in vitro was investigated in wild-type and in IRF-1 and IRF-2 knockout mice. In response to 35 mg/kg LPS, IRF-1-, but not IRF-2-deficient mice, exhibited much poorer induction of COX-2 gene expression in both the livers and lungs. In vitro, COX-2 mRNA levels were also poorly induced in IRF-1-deficient macrophages, while IRF-2- deficient macrophages exhibited higher levels than in normal macrophages. IRF-1 and IRF-2 were confirmed to activate and repress expression of the COX-2 promoter, respectively, in a transient transfection system and the role of specific DNA binding sites confirmed by site-specific mutagenesis. Collectively, these data provide evidence for an important role for IRF-1 in vivo and in vitro and for IRF-2 in vitro in the regulation of COX-2 expression by LPS. PMID- 12537701 TI - Methadone dose and post-mortem blood concentration. AB - The relationship of methadone dose with post-mortem blood concentration was investigated using data collected from 1994 coronial cases in the Australian state of New South Wales. Data on 31 subjects were summarized using linear regression. The weight-adjusted methadone dose, gender, methadone maintenance treatment status and its interaction with adjusted-dose were all significant predictors of post-mortem blood methadone concentration. Data on the death of a young man from the toxic effects of three daily doses of 30 mg methadone are used to give an example of a pair of observed (0.74 mg/l) and predicted (0.48 mg/l) post-mortem blood concentrations. The estimated post-mortem blood concentration for male maintenance patients is at least twice the trough plasma levels estimated from previously published studies of living maintenance patients. The estimated post-mortem blood concentration for female maintenance patients is at least three times the estimated trough level of living subjects. We conclude that post-mortem methadone redistribution is probably the principal cause of the observed differences between males and females in post-mortem blood concentrations and the differences between estimated concentrations for living and deceased subjects. PMID- 12537700 TI - Potential impacts on the incidence of fatal heroin-related overdose in Western Australia: a time-series analysis. AB - In response to the rising concerns about the rate of heroin-related fatalities, overdose prevention campaigns, run by both users' organizations and government agencies, have been implemented in a number of states across Australia. In Western Australia (WA) in mid-1997, various overdose prevention initiatives were implemented. These included the implementation of a protocol limiting police presence at overdose events; the commencement of naloxone administration by ambulance staff; and the establishment of the Opiate Overdose Prevention Strategy (OOPS) which provided follow-up for individuals treated for overdose in emergency departments. This paper reports the results of a multiple linear regression analysis of 60 months of time-series data, both prior to and following the implementation of these interventions, to determine their impact on the number of fatal heroin overdoses inWA. The model employed in the analysis controlled for changes over time in proxy indicators of use and community concerns about heroin, as well as market indicators. The results suggest that, although the interventions implemented have managed to reduce the expected number of fatalities, they have become less successful in doing so as time passes. This has implications for both existing and potential interventions to reduce fatal heroin related overdose. PMID- 12537702 TI - Characteristics, retention and readmissions of opioid-dependent clients treated with oral naltrexone. AB - The aims of this study were to examine the retention rates of opioid-dependent clients treated with oral naltrexone and identify factors that influence retention in treatment of 981 opioid-dependent clients at a public out-patient clinic in Perth, Western Australia. The average retention period for all clients was 9.0 weeks. The factors associated with longer retention were being employed and referral source. Clients who were employed stayed significantly longer in treatment than unemployed clients. Clients referred from a private clinic were retained in treatment significantly longer than those referred from other sources (X = 10.3 vs. 5.9 weeks). While the majority (80.8%) had one admission to naltrexone treatment, 19.2% presented for readmission, some on three or more occasions in the study period. The median period between the end of the first episode of treatment and commencement of the second was 15.6 weeks. The median period between the end of the second episode of treatment and commencement of the third was 11.4 weeks. Those employed had a higher probability of being retained longer in treatment than those who were unemployed in subsequent treatment episodes. Clinicians should expect that initial retention in naltrexone is likely to be relatively short, and that a substantial proportion of clients will represent for further treatment. PMID- 12537703 TI - The emergence of potent forms of methamphetamine in Sydney, Australia: a case study of the IDRS as a strategic early warning system. AB - A striking finding of the Illicit Drug Reporting System (IDRS) in recent years has concerned the emergence in Sydney of a number of different forms of potent methamphetamine. This paper demonstrates the operation of the IDRS in detecting and documenting an increase in both the availability and use of these forms of methamphetamine in Sydney, and Australia more widely, since 1998. Data from different components of the IDRS are utilized to propose that there are currently at least three forms of methamphetamine available in Sydney that are considered by the market to be distinct commodities: methamphetamine powder ('speed'), base methamphetamine ('base') and crystalline methamphetamine ('ice'/'shabu'/'crystal meth'). Base and ice are more potent forms of methamphetamine than speed, and international experience, along with preliminary data obtained from the IDRS, suggest that their use is associated with harms in a number of domains. PMID- 12537704 TI - Diversity of substance use in eastern Arnhem Land (Australia): patterns and recent changes. AB - The objective of this study was to describe patterns of substance use among remote Aboriginal community populations. The setting was the eastern Arnhem Land ('Miwatj') region of the Northern Territory's (NT) 'Top End', with a population of 4217 Aboriginal people over 15 years of age using a cross-sectional description and comparison. Sample 1 (n = 689) from the region used data from health-worker consensus classification of kava, alcohol, tobacco, petrol and cannabis use. Sample 2 (n = 101) from one community used self-reported use, age at commencement, duration, amounts consumed and expenditure. In 1999 (sample 1), 46% of males and 18% of females were kava users, alcohol: 53% males, 12% females, tobacco: 68% males, 65% females, and cannabis: 31% males, 8% females. Less than 5% sniffed petrol. In one community in 2000, 39% males and 20% females reported using cannabis during the previous month. In this community between 1999 and 2000, the proportion of current kava users among men declined (77-52%, p = 0.015) with a tendency in women for a decrease in the proportion of tobacco users (87 69%, p = 0.096). The increase in the proportion of cannabis users in men (21-39%, p = 0.068) was not statistically significant. However, in women the increase was significant (0-20%, p = 0.013). Gross expenditure on tobacco and kava were similar in 2000: both greater than cannabis and alcohol. Median years used ranged from 4 years for cannabis and 20 years for tobacco. The data supported anecdotes of a recent rise in cannabis use, especially in women. Kava use declined in men. Tobacco use patterns in women may have been changing. Average per capita consumption of alcohol was low compared with other 'Top End' areas. Such varied and dynamic substance use patterns pose challenges for research and policy. PMID- 12537705 TI - Perceived and self-reported licit and illicit drug use among fishing industry workers on the mid-north coast of Western Australia. AB - This paper describes the perceptions of boat owners and regional health workers about the use of alcohol and illicit substances within the West Australian mid west coast fishing industry. It also reports on a survey conducted among fishing industry workers concerning their consumption of alcohol and illicit drugs while at sea and in port. Boat owners and health workers perceived that cannabis and alcohol were the most commonly used substances, a perception borne out by the results of the survey. While the levels of illicit drug use for this group were lower than those reported in the fishing industry in an eastern Australian state (Queensland), they were nevertheless considerably higher than those recorded for the general population aged up to 35 years in a National Drug Survey. Cannabis was indeed the most frequently used illicit drug and injecting drug use was minimal. Alcohol use was particularly high and binge drinking described frequently. The results are compared with other industry findings and discussed in terms of occupational health and safety. Recommendations are suggested for future prevention programmes. PMID- 12537706 TI - Recent trends in benzodiazepine use by injecting drug users in Victoria and Tasmania. AB - To address the lack of data on patterns of benzodiazepine use among injecting drug users (IDU) in Victoria and Tasmania, convenience samples of 152 Melbourne and 100 Hobart IDU were recruited from needle and syringe programme outlets and administered a structured survey on patterns of benzodiazepine use, injection related health problems and drug use history. Most respondents had used benzodiazepines during the preceding 6 months, and more than one-third (Melbourne 36%, 95% CI, 28-44; Hobart 37%, 95% CI, 27-47) had injected benzodiazepines during this period. Diazepam was the preferred benzodiazepine for those using orally, while intravenous benzodiazepine users preferred to inject temazepam. Benzodiazepine injection for Melbourne IDU was related to greater levels of injection-related health problems. Patterns of benzodiazepine use amongst Melbourne and Hobart IDU are different to that in other Australian jurisdictions, with available data suggesting that prevalence of injection may be increasing. Ongoing monitoring of benzodiazepine injection, together with in-depth studies of supply characteristics and health impacts in jurisdictions where significant trends are detected is needed. Consideration of regulatory, supply, education and training options for the prevention of benzodiazepine injection is also indicated. PMID- 12537707 TI - The effectiveness of cannabis crop eradication operations in New Zealand. AB - At present the only information available on the effectiveness of the cannabis crop eradication programme in New Zealand is the total number of cannabis plants destroyed each year. These figures can only provide a very crude measure of the effectiveness of these operations. A better measure would be the percentage of total cannabis production destroyed--known as the drug seizure rate. This paper calculates the seizure rate of the cannabis crop eradication programme in New Zealand using the amount of cannabis reported consumed in the Alcohol and Public Health Research Unit's (APHRU) National Drug Survey. The seizure rate for the 1998 programme is calculated to be 26-31%. This compares favourably with drug seizure rates reported in other countries. The effectiveness of the cannabis crop eradication programme, and its apparent modest share of the total cannabis control budget, raises some intriguing questions about the role an expanded crop eradication programme could play in a future cannabis control strategy. PMID- 12537708 TI - The feasibility and acceptability of introducing brief intervention for alcohol misuse in an urban aboriginal medical service. AB - We report on the feasibility and perceived acceptability of brief motivational interviewing for hazardous alcohol use in an urban Aboriginal health service. General practitioners (GPs) were trained in brief motivational interviewing, and health workers in other aspects of the intervention. Screening was initially carried out using the AUDIT, but subsequently reduced to two simple questions. Information was obtained through a combination of participant observation by the study team, ongoing ad hoc review and feedback from staff, periodic group meetings, and one-on-one interviews with health workers and GPs. The AUDIT was felt to be intrusive and some questions were poorly understood. Brief intervention seemed to be culturally appropriate, but barriers to wider administration included lack of time and the complexity of patients' presenting health problems. As a result of the research there was an increase in general awareness and acceptability of addressing alcohol issues at the health service. This study raises a number of issues that both support and threaten the wide implementation of brief intervention in urban Aboriginal primary care settings. PMID- 12537709 TI - Under the radar--how the tobacco industry targets youth in Australia. AB - Tobacco consumption has been declining in Australia since the 1970s when controls on advertising were first introduced. Since this time, legislation has been progressively introduced, severely restricting tobacco advertising and promotion in the mainstream media. This has resulted in limited opportunities for the tobacco industry to reach new smokers, particularly young people. This paper outlines marketing strategies used by tobacco companies and their advertising agencies to reach this group; it examines how the industry exploits loopholes in current legislation and identifies new promotional opportunities. Increasingly, the industry has targeted young people through film, dance parties, nightclubs, fashion shows, e-mail and the internet. The industry is also capitalizing on promoting pack design elements and enhancing them through event promotion. Unless restrictions on tobacco marketing and promotion are comprehensive they undermine the effectiveness of those already in place and will continue to be exploited by the tobacco industry. The recent announcement by the Federal government to reassess the current legislative restrictions in light of these new marketing trends is welcome. The removal of all incentives to promote tobacco products, including imagery associated with the pack and its design, is essential in removing one of the key factors influencing the uptake and prevalence of smoking in youth. PMID- 12537710 TI - Magistrates' Early Referral into Treatment (MERIT): preliminary findings of a 12 month court diversion trial for drug offenders. AB - The purpose of this paper is to present a description and preliminary findings of a 12-month trial of a Local Court diversion programme, called MERIT for Magistrates' Early Referral into Treatment. The aim of MERIT is to divert eligible drug offenders to treatment and rehabilitation services. A total of 172 offenders were assessed and 131 entered the programme. The sources of referral were court (58%), police (17%) and self (10%). Main problem drugs were heroin (57%), cannabis (21%) and amphetamines (11%). The majority (85%) had previous convictions and 50% had been in jail. At the end of the trial period one-third (33%) completed the programme and one-third (33%) remained in treatment. Main treatment interventions were case management and out-patient counselling, detoxification, residential rehabilitation and methadone maintenance. Police records showed that of the original 43 (33%) graduates only six had come to police notice, mainly for relatively minor offences. Early acceptance and preliminary results has led to an expansion of the MERIT programme across New South Wales. With the rapid expansion of drug courts and diversion programmes across Australia, descriptive studies are useful to provide beneficial data to assist policy makers and service providers to develop programmes. PMID- 12537711 TI - From harm reduction to human rights: bringing liberalism back into drug reform debates. AB - This Harm Reduction Digest marks a return to theoretical discussions about harm reduction. Andrew Hathaway notes that harm reduction seldom articulates or acknowledges the moral foundation on which it might build to affect meaningful changes in policy. He argues that despite the rhetorical strengths of empiricism, an openly liberal, human rights orientation imbues rational argument with the principles needed to sustain pragmatic drug reform solutions. Liberalism, with its norms of social tolerance and respect for civil liberties, is presented here as key to the future development of harm reduction discourse as a way of advancing human rights themes in contemporary drug policy debates. PMID- 12537714 TI - Foreword. PMID- 12537712 TI - Administration of a lighter-coloured methadone liquid. PMID- 12537715 TI - Syphilis epidemic in Slovenia influenced by syphilis epidemic in the Russian Federation and other newly independent states. AB - In 1994 an 18-fold increase in the annually reported incidence rate of early syphilis (to 1.81 cases per 100,000 total population) in comparison to the preceding year was observed in Slovenia. The reported incidence rates remained elevated for a period of five years. The possible link between the evolving syphilis epidemic in the Russian Federation and newly independent States (NIS) during the 1990s and the observed increases in the reported early syphilis incidence rates in Slovenia, was explored. Surveillance data obtained by mandatory reporting of all newly diagnosed syphilis cases between 1994 and 1999 were analysed according to the information on sexual partners, the country of probable source of infection, and occupation. More than half of all reported cases (62%) were directly or indirectly linked to a source of infection abroad. Among these, the majority of cases (73%) were linked to the Russian Federation or NIS. Of these, 68% occurred in males, a high proportion being long-distance lorry drivers. PMID- 12537716 TI - Feasibility of testing for Chlamydia trachomatis in a general population sexual behaviour survey in Slovenia. AB - Our objective was to assess the feasibility of integrating first void urine (FVU) specimens testing for Chlamydia trachomatis genitourinary infection into a general population sexual behaviour survey. A total of 752 randomly selected respondents aged 18 to 54 were enrolled into the survey. Face to face interviewing with self-administered sensitive questions was used. Overall survey response rate was 77.4%. A convenience sub-sample of 83 respondents were invited to provide FVU specimens for confidential testing for C. trachomatis genitourinary infection. Fifty-five complied. This resulted in 66% FVU specimen participation rate among targeted respondents. Two specimens tested positive by Amplicor polymerase chain reaction. High feasibility study overall response rate indicated good acceptability of the survey. It proved feasible to collect FVU specimens for C. trachomatis testing in the small sub-sample. Consequently, we proceeded with integration of testing for C. trachomatis into the ongoing main survey. PMID- 12537717 TI - In gratitude for Dutch medicine, dermatovenereology: an insider, outsider. PMID- 12537718 TI - Significance of sexual route of transmission of hepatitis B and C in Russia. AB - It is generally understood that hepatitis B and hepatitis C may be sexually transmitted. During the last decade there was a sharp growth of hepatitis B and C in Russia. In comparison to 1992 the incidence of hepatitis B in Russia rose two fold and in 1999 there were 43.31 cases per 100,000 of population (in some cities up to 150/100,000 and even more). The incidence of hepatitis C in 1999 (19.31 per 100,000 of population) rose to six times more than in 1994. At the same time there was a dramatic growth in syphilis and other sexually transmitted infections in Russia. The proportion of sexual transmission of hepatitis B virus (HBV) and hepatitis C virus (HCV) compared with other routes of transmission increased. According to the data from Moscow City Centre of Epidemiology during the last two years, up to 40% cases of HCV and HBV were sexually transmitted. The most dramatic growth of registered cases of hepatitis was seen among the sexually active population aged 14-29. Confirmation of the sexual route of transmission of HBV and HCV in teenagers was seen when the results of the study showed various markers of HBV and HCV to be significantly more common among sexually active (n = 45) than sexually inactive (n = 341) teenagers (13.33% vs. 4.39% for HBsAg; 46.67% vs. 12.61% for HBsAg+anti-HBs+anti-HBc; 9.47% vs. 3.95% for anti-HCV, respectively). PMID- 12537719 TI - The natural course of asymptomatic Chlamydia trachomatis infections: 45% clearance and no development of clinical PID after one-year follow-up. AB - The natural course of asymptomatic Chlamydia trachomatis infections in women was studied during one year in a cohort based nested case-control study. Healthy women (n = 744, from four company health services in Amsterdam) with a medical check-up prior to job engagement were included. C. trachomatis-positive women (n = 30, cases) and a randomly selected control group of C. trachomatis-negative women (n = 186, controls) were followed for one year. Urine specimens (at one, six and 12 months) were analysed for the presence of C. trachomatis-DNA and the C. trachomatis-serovars, and questionnaires were filled in. The C. trachomatis prevalence and natural course in relation to demographic and sexual characteristics after one, six and 12 months were studied. The main outcome measures were 1) the prevalence of C. trachomatis using urine specimens; 2) self reported complaints; 3) clinical symptoms reported to the coordinating physicians. The prevalence of asymptomatic C. trachomatis infections was 4% and there was no correlation with demographic and sexual characteristics. The person/year clearance rate was 44.7% per year. None of the C. trachomatis positive women developed clinical symptoms or used C. trachomatis specific antibiotic treatment. Women with or without an asymptomatic infection had the same number of self-reported urogenital complaints during follow-up. In persisting infections twice as many C. trachomatis-serovar E infections were detected as compared to clearing infections. Our findings showed that almost half of the asymptomatic C. trachomatis infections in women cleared during one year of follow-up and none developed clinical pelvic inflammatory disease (PID), which is a much lower figure than previously suggested. Therefore these data are important for cost effectiveness calculations in screening programmes for asymptomatic C. trachomatis infections. PMID- 12537720 TI - Prevalence and persistence of asymptomatic Chlamydia trachomatis infections in urine specimens from Danish male military recruits. AB - Danish male military recruits (n = 388) were included in a follow-up study to investigate the prevalence and persistence of asymptomatic Chlamydia trachomatis infections. Urine specimens were collected at enrolment and after approximately six months. C. trachomatis was detected by polymerase chain reaction (Amplicor, Roche). Questionnaires were filled out concerning sexual behaviour and clinical symptoms. The prevalence of asymptomatic C. trachomatis in Danish male military recruits was 4.6% (18 out of 388). From five C. trachomatis-positive men no follow-up sample was obtained. From the remaining 13 C. trachomatis-positive men four (31%) were treated for C. trachomatis between the two visits (outside the study protocol). Of the remaining nine men, one cleared the infection and eight men (89%) had a persistent infection. The number of lifetime sexual partners was associated with the presence of C. trachomatis at enrolment. Although based on small numbers, this follow-up study shows, in contrast to women with asymptomatic C. trachomatis infections, a high percentage of C. trachomatis persistence in asymptomatically infected males. PMID- 12537721 TI - Sensitivity and specificity of three new commercially available Chlamydia trachomatis tests. AB - In order to determine the value of new Chlamydia trachomatis (Ct) specific tests for routine serological diagnosis of Ct infections, we evaluated several commercially available assays (C. trachomatis enzyme immunoassay (EIA), Labsystems (CtL); SeroCT, Savyon (CtS); pELISA, Medac (CtMp)) in various study populations. The prevalence of C. trachomatis-specific IgA antibodies in a blood donor population (n = 443) as determined by the peptide based tests CtL, the CtS and the CtMp was 5%, while for IgG antibodies this was 6% (CtL and CtS) and 12% (CtMp) respectively. Prevalence was negatively correlated with age, concording with C. trachomatis specificity. None of the three tests showed significant titre rises in serum samples taken from patients with a proven infection of Chlamydia pneumoniae (n = 22), indicating species-specificity for all three tests. In patients with a polymerase chain reaction proven (n = 324) Ct infection, 75%, 70% and 68% were positive for IgG and 45%, 38% and 47%, positive for IgA as determined by the CtMp, CtL and CtS respectively. We conclude that the new synthetic peptide-based EIA tests are able to detect species-specific Ct antibodies, which are strongly correlated to (active) infection. PMID- 12537722 TI - Tubal factor pathology caused by Chlamydia trachomatis: the role of serology. AB - One of the causes of infertility in females is tubal pathology as a result of pelvic inflammatory disease (PID) caused by Chlamydia trachomatis (Ct). Diagnosis and identification of patients is hampered by the lack of rapid, easy, sensitive and specific methods. The introduction of Ct-specific enzyme-linked immunosorbent assay (ELISA) tests, based upon synthetic peptides may subsequently increase the sensitivity and specificity for the detection of tubal factor infertility caused by Ct. In order to determine the value of these tests for serological diagnosis of Ct infections, we evaluated several commercially available assays (C. trachomatis enzyme immunoassay (EIA), Labsystems (CtL); SeroCT, Savyon (CtS); pELISA, Medac (CtMp); and a reference assay rELISA, Medac (CtMr)) in two study populations. The first group consisted of 134 female patients with infertility problems. Tubal factor infertility was observed in 85 of these patients (63%). A higher % positivity was found for Ct-specific IgG for the CtL, CtS and CtMp, 41% vs 10%, 57% vs 18% and 55% vs 25% respectively as compared to patients with infertility due to other problems. A similar trend was observed for Ct-specific IgA. The specificity of Ct-specific IgA and IgG in this patient group varied between 92 to 98% and 76 to 90%, respectively. The second group consisted of 107 consecutive gynaecology patients with fertility problems or suspected PID. In this particular patient group, the specificity of the peptide based tests were around 80% and 90% for Ct-specific IgA and 75% and 85% for Ct-specific IgG, respectively. The negative predictive values exceeded 90%, while the positive predictive values varied from 30% to 47% for Ct-specific IgA and was around 30% for Ct-specific IgG. Testing Ct-specific IgG had no additional value above Ct specific IgA alone. We conclude that the new synthetic peptide-based EIA tests are able to detect species-specific antibodies, which are correlated to (active) infection, and that in particular IgA may be useful in the serodiagnosis of tubal factor infertility caused by C. trachomatis, and will contribute in simplifying the work-up in patients with infertility. PMID- 12537723 TI - Post-exposure prophylaxis. AB - The mean risk of acquiring HIV after an occupational exposure, injecting drug use or sexual exposure varies from < 0.1 to 3%. A high plasma HIV-RNA of the source increases the risk of each of the exposures. Other factors, such as the volume of the inoculum involved to which the individual was exposed, other sexually transmitted diseases and ruptures of mucous membranes are associated with a higher risk of HIV transmission. Based on the calculated risk, post-exposure prophylaxis (PEP) should be recommended. In the Netherlands, prescription of PEP in the occupational setting is a standard procedure and has proved to be feasible. This was associated with a high percentage (62%) of mild and reversible toxicity and a small percentage (2%) of serious adverse events related to antiretroviral drugs, i.e. nephrolithiasis (due to indinavir) and toxic hepatitis (due to nevirapine). In The Netherlands so far no HIV-seroconversions have been recorded after an occupational accident. PMID- 12537724 TI - Experience with the new German hepatitis B vaccination strategy since 1995. AB - In Germany general hepatitis B (HB) vaccination for newborns and adolescents was introduced in all federal states following the recommendation of the STIKO (Permanent Vaccination Commission) of 1995. In 1998 serological studies of the German National Health Survey showed that at least 9.8% in the age group of 18 to 19 years had been vaccinated against HBV infection. By 1996 the vaccine doses sold for children rose dramatically and started to reach a plateau of approximately five million per year in 1997. Data from the kassenarztliche Vereinigung of the Oberpfalz region in Bavaria also indicate that the new policy started to be implemented in 1996. At school entry, however, in 1997 only 10% of the children in seven West German states showed serological evidence of HB vaccination coverage. According to observations of virologists, paediatricians and public health experts the current acceptance of the HB vaccination recommendations is 80%-90% in children below the age of six years but only 30% to 40% in adolescents. To achieve high HB vaccination coverage rates in Germany more rapidly a modern surveillance system providing detailed data about vaccination coverage in the different age and population groups is needed. Based on those data additional targeted vaccination strategies for those that can only be contacted by the traditional health care system, such as teenagers and vulnerable groups, should be developed, involving the public health service, local communities and other agencies. PMID- 12537725 TI - Vaccine candidates in STD. AB - Sexually transmitted diseases (STDs) are caused by organisms that infect the mucosal surfaces of the genitourinary tract. In spite of its public health importance, current STD vaccine research lags behind work against pathogens that target another mucosal region, the respiratory tract. In the latter case, live attenuated viral vaccines, killed whole-cell bacterial vaccines, subunit/protein bacterial vaccines, and bacterial polysaccharide vaccines have been enormously successful. To move STD vaccine research forward, complex issues must be resolved. Those include selection of an appropriate antigen (e.g. scientific feasibility and intellectual property rights), the manufacture of the vaccine (e.g. delivery systems, formulation processes, and production steps), and the appropriate public health approach (e.g. medical indications and marketing aspects). Particular scientific problems have delayed STD vaccine development, like incomplete attenuation (human herpes simplex virus type 2), accentuated immunopathology (Chlamydia trachomatis), poor immunogenicity (Treponema pallidum), and broad antigenic heterogeneity (Neisseria gonorrhoeae). Nevertheless, efforts continue with the use of protein antigens: for example, the haemolysin toxoid of Haemophilus ducreyi; the major outer membrane protein(s) of N. gonorrhoeae and C. trachomatis; the glycoprotein D of human herpes simplex virus type 2; and the proteins E6 and E7 of human papilloma virus. It may be predicted that eventual STD vaccines (administered either for prophylaxis or for therapy) will use approaches that include (1) live-attenuated viruses, (2) subunit proteins or inactivated whole organisms given with mucosal adjuvants or with cellular immune response adjuvants, and (3) DNA plasmids expressing the vaccine antigen. PMID- 12537726 TI - The sins of the fathers--Africans with HIV infection in London; lessons for others? AB - Many European countries have taken in immigrants from sub-Saharan Africa. The reasons for this are discussed and the particular problems experienced by HIV infected Africans in London, and the approach to their care at St Thomas' Hospital, is delineated. PMID- 12537727 TI - Do Dutch dermatologists care for venereology? An inquiry among all non-academic dermatologists in The Netherlands. AB - The financing of STD outpatient clinics in The Netherlands is currently undergoing structural changes. Because these changes also have implications for the infrastructure of STD care as a whole, the STD committee of the Dutch Society for Dermatology and Venereology (STD committee NVDV) and the National Society of Municipal Health Services (GGD-Nederland) are currently exploring the possibilities and feasibility of intensified regional collaboration between Municipal Health Services (MHSs) and dermatologists. However, for fruitful collaboration it is essential that a substantial number of dermatologists has an interest in STD care. Therefore, the STD committee NVDV has conducted a structured survey in order to study the support of Dutch dermatologists for such a regional collaboration. In this paper, the results of the survey are presented. It appears that the majority of Dutch dermatologists is (still) interested in STD, and although a minority currently collaborates with local MHSs on a regular basis, a large group is willing to do so in the future. We conclude that the majority of dermatologists in the Netherlands (still) cares for venereology and that there is a sound basis for a fruitful cooperation with MHSs. PMID- 12537728 TI - The international community--what are the problems? What are we doing? Where do we go from here? PMID- 12537729 TI - Urinary tract infection in patients with acute non-gonococcal urethritis. AB - In young sexually active male patients, clinical differentiation between non gonococcal urethritis (NGU) and urinary tract infection (UTI) can be difficult. UTI as a cause of NGU has been suggested before, but the prevalence of UTI among acute NGU patients has not been evaluated. We conducted a prospective cross sectional study of 156 male patients with clinical features of acute urethritis. The prevalence of UTI among acute NGU patients was 6.4% (CI: 1.5-11.3%). There was a possible association between UTI and NGU but this was not statistically significant. The sensitivity and specificity for combining leucocyte esterase and nitrite tests were 83.3% and 89.8%, respectively, with a negative predictive value of 98.8%. We conclude that a mid-stream urine specimen should routinely be collected in patients with symptoms of urethritis. This should be analysed by dipsticks incorporating nitrite and leucocyte esterase tests. Those with a positive dipstick should then be confirmed by culture. PMID- 12537730 TI - What is new with antibiotic-resistant gonorrhoea in Newcastle, England? AB - Neisseria gonorrhoeae isolates that were resistant to ciprofloxacin and/or penicillin were analysed to investigate the escalating problem of antibiotic resistant gonorrhoea in the north east of England. Opa-typing (outer membrane opacity protein) was carried out on isolates resistant to ciprofloxacin and of nutrient nonrequiring (NR) auxotype. In the year 2000 there were 265 cases of gonorrhoea, of which 44 (16.6%) were resistant to penicillin and 12 (4.5%) were resistant or had reduced sensitivity to ciprofloxacin (with only four of these acquired outside the UK). Three (7.5%) of the non-beta-lactamase penicillin resistant isolates were imported from abroad. By Opa-typing of ciprofloxacin resistant strains, one pair of the isolates was similar, two were unique and one was similar to the Oldham/Rochdale outbreak strain described early in 2000. This marked increase in the prevalence of indigenous ciprofloxacin resistance requires continued surveillance and may soon necessitate an alteration in our first line treatment. PMID- 12537731 TI - A survey of non-consultant career grade doctors in genitourinary medicine: their knowledge of guidelines and their access to information technology 2001. AB - Non-consultant career grade (NCCG) doctors in genitourinary medicine (GUM) perform a large proportion of the clinical work. To ensure quality of service delivery to patients, it is essential that these doctors keep up to date. Seven hundred questionnaires were sent to NCCGs in the UK to evaluate their knowledge of national guidelines and access to information technology. A summary of the 224 replies (31% response) is presented. Knowledge of guidelines (76%-86%) and access to the Internet at work (39%-44%) varied according to the number of sessions worked in GUM per week and by grade of doctor. Knowledge of relevant websites was poor, ranging from 40%-54% for the Medical Society for the Study of Venereal Diseases (MSSVD) website and 21%-39% for the Association of Genitourinary Medicine (AGUM) website. This survey highlights areas for concern, especially with regard to NCCGs who work three or less GUM sessions per week. PMID- 12537732 TI - Coitarche and care: does experience of the 'looked after' system affect timing of a woman's sexual debut? AB - Women with an early coitarche may be less connected to home than those with a later coitarche and are more at risk of sexually transmitted infections (STIs). Children in care or the 'looked after' system feel less connected. We investigated whether women ever in care (WEIC) differed from women never in care (WNIC) in terms of coitarche and STIs using self-reported data from women attending Genitourinary Medicine clinics. Of the 336 women participating 17 women had experienced care, of whom 13 (76.5%) had coitarche below 16 years of age; of these five (38.5%) had used a condom at coitarche and three (60%) with genital infections had gonorrhoea, chlamydial infection or pelvic inflammatory disease. This compared with 112 (35.1%), 70 (62.5%) and 29 (40.8%), respectively for WNIC. WEIC are more likely to have early coitarche; if this is below 16 years subsequent genital infections are more likely to be fertility-threatening. PMID- 12537733 TI - A behaviour sentinel surveillance for female sex workers in the Social Hygiene Service in Hong Kong (1999-2000). AB - Behavioural serial sentinel surveillance survey for female sex workers (FSWs) was carried out in the Government Social Hygiene Clinics in Hong Kong during 1999 and 2000. A total of 1366 and 1451 FSWs participated in the questionnaire survey respectively. The results showed that the majority of FSWs attending the clinics were of Chinese ethnicity (83.8%, 87.6%) while Thai (13.0%, 9.49%) and Filipino (2.1%, 2.7%) were the main foreign ethnic groups. The karaoke night club was the most frequent reported workplace (32.0%, 32.6%) followed by a club (8.9%, 11.7%) and free-lancing (8.6%, 9.0%). The proportion of FSWs who always used condoms in vaginal sex with commercial sex partners was 70.9% and 75.0% respectively (P=0.038). Compared with previous results, a rising trend of condom use by FSWs in commercial sex was observed. However, there was a relatively low condom use during commercial oral sexual intercourse (30.7%, 36.6%) and sex with their regular sex partners in all forms of sexual activities. Client dislike is the main attributed factor for not wearing condoms. The point prevalence of both infectious syphilis and HIV infection was 0.1% in both years. These data are useful for planning, resource allocation and evaluation of outreach activities and STI services targeting at FSWs and their clients. They provide a sampling framework for further surveillance. PMID- 12537734 TI - Opportunistic chlamydia screening; should positive patients be screened for co infections? AB - This study examines the requirement for testing patients for other sexually transmitted infections (STIs) and bacterial vaginosis (BV) when diagnosed with genital chlamydia during opportunistic screening. Data were collected on all patients participating in the Department of Health chlamydia screening pilot study in Portsmouth. One thousand two hundred and forty-five women and 490 men with genital chlamydia were seen in Portsmouth genitourinary medicine (GUM) department. Of the women screened in GUM, 28% had coexisting STIs and 21% had BV. The corresponding figures for those initially screened in the community were 4% and 17%. An increased number of female sexual partners of male patients (76%) and male partners of female patients (55%) of the GUM group had co-infections; 58% of male partners from the community group had another STI. The increased morbidity associated with these infections warrants screening of all patients with chlamydia for other STIs and BV. PMID- 12537735 TI - Antimicrobial susceptibility of Neisseria gonorrhoeae strains isolated in Nuuk, Greenland during 1998-1999. AB - Sixty-one gonococcal strains isolated in Greenland during 1998-1999 were tested locally for susceptibility to penicillin and ciprofloxacin by a disc diffusion method (Rosco) and at the reference laboratory in Copenhagen by the agar dilution method and the E-test, showed that more than 60% of the strains were less susceptible or resistant to penicillin (minimum inhibitory concentration [MIC] > or = 0.25 mg/L), indicating that penicillin should not be used as a first line drug for the treatment of gonorrhoea in Greenland. The Rosco disc diffusion method only identified 8% of strains as less susceptible to penicillin. Ciprofloxacin can still be used as a first line drug for treatment of gonorrhoea in Greenland since decreased susceptibility was only seen in one imported strain. However, the decreased susceptibility in this strain was not identified by the Rosco disc diffusion test, therefore the future local surveillance of antibiotic resistance in Neisseria gonorrhoeae will be based on MIC determinations. PMID- 12537736 TI - AIDS awareness and sexual behaviour in a high HIV prevalence area in rural northern Kwazulu-Natal, South Africa. AB - The objective was to assess AIDS awareness and sexual behaviour in a rural South African community with a high HIV prevalence. One hundred clinic attenders underwent a structured interview using a standard questionnaire. Although the 64 female and 36 male patients, mean age 22 (range 13-45), had good knowledge of AIDS-related issues, only 50 perceived HIV/AIDS as a common problem. Of the 75 patients who were sexually active only 30 (40%) used condoms (men 16; 55% vs women 14; 30%, P=0.033) despite being better informed about the protective effect of condoms (active 61; 81% vs abstinent 14; 56%, P=0.011). More men than women admitted to multiple sexual partners (17; 47% vs 7; 11%, P<0.0001). In conclusion, despite a high level of awareness of HIV/AIDS issues, self-perceived risk was low, condom use was infrequent and especially men continued to have multiple sexual partners. Awareness has yet to translate into reduction of risk behaviour. PMID- 12537737 TI - Prevalences of and risk factors for sexually transmitted diseases among Japanese female commercial sex workers in middle- and high-class soaplands in Japan. AB - A cross-sectional study among Japanese female commercial sex workers (CSWs) working in soaplands (massage parlours with baths) was conducted between July 1999 and June 2000. The study subjects included 135 CSWs aged from 20 to 36 years. A questionnaire included sexual characteristics in addition to real name, working name, and date of birth. We serologically or bacteriologically confirmed prevalences of HIV-1, HIV-2, hepatitis B virus (HBV), hepatitis C virus (HCV), Chlamydia trachomatis, Neisseria gonorrhoeae, syphilis, and trichomonads. Anti HCV positive was 0.7%. Although anti-HBs positive was 15.6% no HBsAg was confirmed. Seroprevalence of C. trachomatis was 8.9% and of syphilis was 4.4%. The other STDs studied were not detected. A statistically significant difference was observed between middle-class CSWs and high-class ones about condom use (P<0.01). Prevalences of STDs among Japanese female CSWs working in middle- and high-class soaplands were evidently lower than those reported previously. PMID- 12537738 TI - HIV epidemiology in Morocco: a nine-year survey (1991-1999). AB - The first case of HIV/AIDS in Morocco was notified in 1986. The Moroccan Ministry of Health has set up a system of management of HIV/AIDS cases and surveillance of the epidemic. Between 1991 and 1999, we studied 12,981 sera from patients in different Moroccan regions, who attended with high risk and/or symptoms of HIV/AIDS. The overall prevalence of positive HIV serology was 5.04% (4.67-5.44%). This rate was higher compared to blood donors which is 0.02%. From 1993 to 1998, the prevalence was around 4-5% (2.73-6.26%) then increased to 8.06% (6.89-9.36%) in 1999. Retrospective analysis of available epidemiological data have shown that the highest rate was observed in the South of Morocco with 12.23% (11.10-13.44%). Of the HIV positive cases, 72.22% of seropositives were infected heterosexually, 11.11% by intravenous drugs use, 7.41% by homosexuality, 4.63% by bisexuality and 4.63% and 0.62% by vertical transmission. The median age of infected persons was 33.3 years. Among documented HIV-positive individuals, 53.74% were aged 30-39 years, 21.43% were 20-29 years and 17.69% were 40 years or more. In this study, among HIV-positive patients 42.17% have loss of weight with fever and diarrhoea represent and 35.54% have loss of weight with fever, diarrhoea and opportunistic infections. The present work shows that the HIV infection increased significantly in 1999. Effective measures to control this trend should be implemented soon before the situation worsens. PMID- 12537739 TI - Which patients attending genitourinary medicine clinics have HIV tests? AB - To identify factors associated with uptake of HIV testing a questionnaire was given to patients attending a GUM clinic over a three-week period. One hundred and twenty (69.4%) of 189 patients accepted and 53 (30.6%) refused testing. Variables associated with having a HIV test were: being tested previously (P=0.045), given a leaflet about testing (P=0.001), told about the window period (P=0.006), told about availability of counselling (P=0.030), given insurance advice (P=0.014), and a past history of sexually transmitted infections (P=0.044). Most patients perceived a low risk of being HIV positive (n=143, 75.7%) with no difference between those accepting or declining testing. The principal reason for testing was a check-up, and for refusal was a lack of perceived risk. Patients who are well informed about HIV testing are more likely to accept a test. PMID- 12537740 TI - The diagnosis of hepatitis C in a semi-rural genitourinary medicine clinic. AB - A retrospective casenote study was conducted to examine the risk factors for patients who were diagnosed as hepatitis C (HCV)-positive (between 1999 and 2001) in a semi-rural genitourinary medicine (GUM) setting in Staffordshire, UK. There was a remarkable escalation in the number of reported HCV-positive cases, year on year, in our study. The majority of the positive cases (20/21) gave a history of previous intravenous drug use (IVDU). The incidence of other sexually transmitted infections were reportedly high in our HCV-positive patients. The increasing number of reported HCV-positive cases in the GUM clinic of the semi-rural Staffordshire setting, may reflect a national pattern that needs further investigations. We advise that HCV serology should be offered to GUM clients (who have a history of IVDU), and to their sex partners. They should also be advised to take tests to exclude other STDs. The case for offering HCV serology as a routine test for patients who request 'the exclusion of STDs' is still undecided. PMID- 12537741 TI - Female genital mutilation--case report and discussion. PMID- 12537742 TI - Human bite-induced penile ulceration: report of a case and review of literature. AB - Human bite on genitalia is a rare cause of penile ulceration, but is increasingly being reported, probably due to the increasing frequency of orogenital sex. The great morbidity associated with it brings it under the category of high-risk bite wounds, similar to those on hands, feet and joints. We report a case and review the literature on human bite-induced penile ulceration. PMID- 12537743 TI - Oral donovanosis. AB - A 23-year-old male presented with oral ulcers. Tissue smear showed Donovan bodies. The patient was treated with oral tetracycline. PMID- 12537744 TI - Periurethral abscess complicating male gonococcal urethritis treated by surgical incision and drainage. AB - Gonococcal urethritis in a heterosexual male complicated by periurethral abscess and its treatment with antibiotics and surgical drainage is presented. PMID- 12537745 TI - A comparison of demographic details, source of referral, and spectrum of infection in patients under 16 years attending Singleton Hospital Genitourinary Medicine Department in 1997 and 2000. PMID- 12537746 TI - Pyrexia of unknown origin in HIV infection and the resurgence of syphilis. PMID- 12537747 TI - Where have all the men gone? Chlamydia infection in men. PMID- 12537749 TI - Physiological consequences of hyperthermia: heat, heat shock proteins and the immune response. PMID- 12537750 TI - Hsp70, an immunological actor playing with the intracellular self under oxidative stress. AB - Heat shock proteins (HSPs) have originally been studied at cellular and molecular levels for their role in resistance to stress. More recently, HSPs have received the attention of immunologists for their ability to stimulate the adaptive and innate immune system. However, the immunological consequences mediated by their expression under stress have not been fully explored. Recent studies on the inducible Hsp70 have promoted the emergence of a new link between oxidative stress and immune response. This view is based on the observation that (i) the stress inducible Hsp70, but not the constitutive Hsc70 co-segregates with immunogenicity of tumour cells in vivo, (ii) the stresses that induce Hsp70 expression also give rise to cellular oxidation, and (iii) an active immune response is a source of oxidative stresses. Here, one explores the hypothesis that, under oxidative stress, Hsp70 is more efficient in discriminating intracellular 'non-self polypeptides' than Hsc70, leading to a more efficient stimulation of the immune system. PMID- 12537751 TI - Cancer immunotherapy: stress proteins and hyperthermia. AB - Heat shock proteins (hsps) can induce anti-cancer immune responses by targeting associated tumour antigens to the immune system. Hsps are not merely carriers of antigen but can also induce maturation of dendritic cells (DCs), resulting in a more efficient antigen presentation. However, improvement of hsp-based vaccines is still desirable if one is to realize their full therapeutic potential. Since the immune system consists of different elements functioning together in a highly integrated way, a combination therapy utilizing important immunomodulators together with hsp-based vaccination may improve therapeutic response. Hyperthermia has been shown to have important stimulatory effects on several cellular and organismal endpoints related to the immune system. This review highlights advantages and disadvantages of various ways of using stress proteins in cancer immunotherapy. It also overviews the interaction of hyperthermia with heat shock protein therapy and the related effects on the host's immune response. PMID- 12537752 TI - The heat shock protein Gp96 links innate and specific immunity. AB - Among other heat shock proteins (HSPs), the ER-resident chaperone Gp96 has been described as a potent tumour vaccine in animal models. A growing list of data underlines that Gp96 triggers both arms of pathogen defence-innate and specific immunity-in a synergistic and most efficient way: It enables specific immune responses by transferring immunogenic peptides that have been acquired in the ER to the MHC class I pathway of antigen presenting cells (APCs). For this, two important features of Gp96 are required. First, its ability to bind immunogenic peptides. Secondly, its acquisition by specialized antigen presenting cells capable of inducing cellular immune responses. Due to specific receptors on the surface of APCs, this uptake from the extracellular space occurs very efficiently and rapidly. Serving the innate branch of immunity, Gp96 unspecifically activates APCs, which then provide a pro-inflammatory cytokine milieu and co-stimulation to cytotoxic T cells. Thus, Gp96 uses all resources of the immune system to trigger cytotoxic T cell responses against associated peptides. PMID- 12537753 TI - Cytokine and adhesion molecule expression in primary human endothelial cells stimulated with fever-range hyperthermia. AB - Migration of blood-borne lymphocytes into lymphoid tissues and sites of inflammation is initiated by vascular adhesion molecules and proinflammatory cytokines. Previous in vivo studies have shown that febrile temperatures dynamically stimulate adhesion in differentiated high endothelial venules (HEV), which are portals for lymphocyte extravasation. This report examines the direct effect of fever-range hyperthermia on the expression of adhesion molecules and cytokines by primary cultured endothelial cells. In both macrovascular (HUVEC) and microvascular (HMVEC) endothelial cells, fever-range hyperthermia (40 degrees C for 6-12 h) did not affect expression of adhesion molecules (ICAM-1, E selectin, VCAM-1, P-selectin, PECAM-1, PNAd, MAdCAM-1), cytokine release (IL 1beta, TNF-alpha, IFN-gamma, IL-6, IL-11, IL-12, IL-13), or chemokine secretion (IL-8, RANTES, MCP-1, MIP-1beta, MIG). This is in contrast to the stimulatory effects of TNF-alpha or 43 degrees C heat shock. However, a novel role for fever range hyperthermia was identified in augmenting actin polymerization in cultured endothelial cells and enhancing the ability of endothelial-derived factors to transactivate the alpha4beta7 integrin lymphocyte homing receptor. These findings provide insight into the tightly regulated effects of fever-range hyperthermia that exclude induction of adhesion in non-activated endothelium of normal blood vessels. Through these mechanisms, it is proposed that febrile temperatures associated with infection or clinical hyperthermia avoid the unproductive exodus of lymphocytes to non-involved extralymphoid tissues while simultaneously promoting lymphocyte delivery to sites of immune activation. PMID- 12537754 TI - Induction of stress proteins in a panel of mouse tissues by fever-range whole body hyperthermia. AB - When exposed to environmental stress, cell survival is supported by the upregulation of stress proteins such as heat shock proteins (HSPs) or glucose regulated proteins (GRPs), which help prevent protein denaturation. To begin to characterize the ability of a physiologically relevant heat exposure to induce stress protein expression, the cerebellum, cerebrum, colon, heart, kidney, liver, lung, lymph nodes, muscle, serum and thymus were extracted from BALB/c mice at various times after fever-range whole body hyperthermia (FR-WBH, 39.5-40 degrees C for 6 h) treatment. The expression of three stress proteins, HSP70, HSP110 and GRP170, was determined in these tissues and serum and compared to constitutive levels in control tissues and serum using Western analysis. Constitutive expression of GRP170 was not affected by FR-WBH in any tissue. In contrast, FR WBH did enhance HSP expression: HSP70 in heart, kidney, lung, lymph nodes and thymus; and HSP110 in lung, lymph nodes and thymus. The lymphoid tissues displayed the most consistent upregulation of both HSP70 and HSP110 upon FR-WBH treatment. The apparent sensitivity of immunologically relevant tissues to FR-WBH may relate to the enhanced immune responses that are observed during febrile temperatures. PMID- 12537755 TI - Heat shock protein 70: role in antigen presentation and immune stimulation. AB - Heat shock proteins (HSP) when released into the extracellular milieu can act simultaneously as a source of antigen due to their ability to chaperone peptides and as a maturation signal for dendritic cells, thereby inducing DCs to cross present antigens to CD8+ T-cells. HSP can also act independently from associated peptides, stimulating the innate immune system. Previous results regarding the activation of NK cells by HSP70 cell surface expression on tumour cells and soluble HSP70 will be further covered elsewhere within this issue. For cross presentation, HSP70-peptide complexes (HSP70-PC) were used from two human melanoma cell lines that differ in the expression of the tumour-associated antigen tyrosinase. Purified HSP70-PC consists of both the constitutively expressed HSC70 and the inducible HSP70. HSP70-peptide complexes purified from tyrosinase positive (HSP70-PC/tyr+) human melanoma cells, incubated with immature DCs, results in the activation of HLA-*A0201-restricted tyrosinase peptide specific T-cells. Receptor-mediated uptake of HSP70-PC by DCs and intracellular transport are required for efficient MHC class I restricted cross-presentation of chaperoned peptides. Demonstration of HSP70-PC mediated cross-presentation of such non-mutated naturally expressed tumour antigens is of special clinical interest with regard to hyperthermia. Tumour regression and improved local control have been shown within clinical phase II/III trials integrating regional hyperthermia combined with radiation and/or chemotherapy in multimodal treatment strategies. According to the proposed concept, local necrosis induced by hyperthermic treatment induces the release of HSPs, followed by uptake, processing and presentation of associated peptides by DCs. By acting as chaperone and a signal for DC maturation, HSP70-PC might efficiently prime circulating T cells. Therefore, upregulating HSP70 and causing local necrosis in tumour tissue by hyperthermia offers great potential as a new approach to directly activate the immune system. PMID- 12537756 TI - Activation of natural killer cells by heat shock protein 70. AB - Intracellular heat shock proteins (HSP) function as molecular chaperones, they support folding and transport mechanisms of other proteins under physiological conditions and following physical or chemical stress. More recently, extracellular localized HSP have been found to play key roles in the induction of a cellular immune response. Either they act as carrier molecules for immunogenic peptides that are presented on Antigen Presenting Cells (APC) to cytotoxic T cells or they themselves act as activatory molecules for the innate immune system. Binding of uncomplexed HSP to HSP-receptors on APC has been found to induce the secretion of inflammatory cytokines. Furthermore, an unusual tumor selective membrane-localization of non-conserved regions of the 72 000 Da HSP (Hsp70) has been found to act as a recognition structure for natural killer (NK) cells. In this review the interaction of NK cells with Hsp70 or peptides derived thereof will be eluciated in more detail. PMID- 12537757 TI - Hyperthermia-regulated immunogene therapy. AB - One of the key milestones that must be reached before gene therapy becomes feasible for clinical cancer treatment is to be able to regulate therapeutic gene expression. This is true for most current cancer gene therapy approaches, since the majority of therapeutic genes are toxic to both tumour and normal tissues. Among the wide array of strategies available for regulating gene expression, hyperthermia represents a unique approach. Hyperthermic regulation of gene therapy is feasible because of the widely conserved heat shock response, which allows therapeutic gene expression to be elevated to thousands of fold higher than background when temperature reaches 3-5 degrees C over physiological temperature (37 degrees C). In addition, because of the long history of experimental research on the use of hyperthermia as an approach for cancer therapy, it is now quite feasible to apply hyperthermia to a number of tumour sites and to achieve temperatures that are sufficient to induce a heat shock response. This review will attempt to discuss the current status of hyperthermia regulated gene therapy, with special emphasis on hyperthermia-regulated immunogene therapy. PMID- 12537758 TI - Targeting HSP70-induced thermotolerance for design of thermal sensitizers. AB - Thermal therapy has been shown to be an extremely powerful anti-cancer agent and a potent radiation sensitizer. However, the full potential of thermal therapy is hindered by a number of considerations including highly conserved heat resistance pathways in tumour cells and inhomogeneous heating of deep-seated tumours due to energy deposition and perfusion issues. This report reviews recent progress in the development of hyperthermia sensitizing drugs designed to specifically amplify the effects of hyperthermia. Such agents might be particularly useful in situations where heating is not adequate for the full biological effect or is not homogeneously delivered to tumours. The particular pathway concentrated on is thermotolerance, a complex, inducible cellular response that leads to heat resistance. This paper will concentrate on the molecular pathways of thermotolerance induction for designing inhibitors of heat resistance/thermal sensitizers, which may allow the full potential of thermal therapy to be utilized. PMID- 12537759 TI - Pituitary adenoma in Carney complex: an immunohistochemical, ultrastructural, and immunoelectron microscopic study. AB - First described in 1985, Carney complex is a rare, heritable disorder featuring abnormal skin pigmentation, cardiac and cutaneous myxoma, melanotic schwannoma of psammomatous type, and endocrine abnormalities, including pituitary adenomas. Patients with the latter present with elevated growth hormone (GH) levels and acromegaly or gigantism. Prolactin (PRL) elevation may also be seen. The authors have investigated 2 resected pituitary adenomas from patients with Carney complex. One, a 19-year-old female acromegalic with elevated GH, IgF-1, and PRL levels, had a mammosomatotroph adenoma immunoreactive for GH and PRL. Ultrastructurally, GH and PRL were present in the same secretory granules. The second patient, a 27-year-old acromegalic, had a sparsely granulated GH cell adenoma that by immuno-electron microscopy revealed GH immunoreactivity only. The lack of morphologic similarity between the 2 adenomas indicatesthat pituitary tumors in patients with Carney complex may not exhibit the same phenotype. PMID- 12537760 TI - Deciduoid mesothelioma: a report of 5 cases and literature review. AB - Deciduoid mesothelioma was first described in young females and in the peritoneum, which led to the suggestion that deciduoid mesothelioma was a distinct subtype with specific clinical and pathologic features. Later reports, however, have shown that this type of mesothelioma may also occur in elderly people and in the pleura. Cases reported in the literature so far are limited, and the disease is not well defined. The authors report the histologic, immunohistochemical, ultrastructural, and clinical findings of 5 cases of deciduoid mesothelioma, and review the literature reports. The results demonstrate that the presence of numerous cytoplasmic intermediate filaments, either dispersed or bundled, appear to be the likely ultrastructural basis for the deciduoid histologic appearance. Twenty-one cases of deciduoid mesothelioma were identified in the literature. Analyses of these 21 cases and the authors' 5 cases showed an age range of 13-78 years (median 53 years) and a slight female predominance (female to male ratio of 1.4:1). Fourteen of 26 cases (54%) occurred in the peritoneum. Seven of 20 patients (35%) had a documented history of asbestos exposure. Fifteen of 20 patients died, with a mean survival time of 7.33 months (range 1-21 months). Five of 20 patients were alive at a follow-up time of 8 months to 5 years. These findings suggest that the so-called deciduoid mesothelioma has some clinical and pathologic features that are dissimilar to mesothelioma in general. Whether it truly represents a pathogenetically distinct variant or merely an expansion of the morphologic spectrum awaits further studies. PMID- 12537761 TI - Infrared microspectroscopic detection of epithelial and stromal growth in the human benign prostatic hyperplasia. AB - Benign prostatic hyperplasia (BPH) is one of the most common diseases in elderly men. It describes an overgrowth of the epithelial and fibromuscular tissues in the human prostate, in which both epithelium and stroma continue to proliferate at different rates. There is lack of suitable techniques for direct evaluation of the biophysical structure of molecular constituents and the proliferative changes in the structure of both compartments at the microscopic level. Reflectance Fourier transform infrared (FT-IR) microspectroscopy was used to pathologically image and examine the structural changes in the proliferation of epithelial and stromal tissues of human BPH. Two successive thin tissue slices were obtained by cutting the tissue with a microtome: one unstained slice was used for FT-IR spectroscopic analysis and the other stained slice was used for position referencing. The result indicates that the amide I maximum for BPH epithelial tissue was near 1,638 cm(-1) and showed its protein secondary structure by a combination of unordered and beta-sheet structures. However, an intense absorption band of amide I was at 1,630 cm(-1) for stromal BPH tissue due to the predominant beta-sheet structure. The higher peak intensity of several IR bands at 1,337, 1,281, 1,238, 1,206, 1,055, and 1,034 cm(-1), assigned to the C-O stretching mode of collagen, was observed in BPH stromal tissue. This suggests that the obvious collagen will be more enriched in the stroma of BPH tissue. PMID- 12537762 TI - Nephrotoxicity of combined treatment with cisplatin and gentamicin in the guinea pig: glomerular injury findings. AB - The drugs cisplatin and gentamicin are given consecutively to various cancer patients suffering from infections. Little information exists about the ultrastructural alterations of kidney glomeruli caused by treatment with these drugs. Renal glomeruli of guinea pigs treated with cisplatin alone and in combination with gentamicin were studied by transmission electron microscopy. The findings revealed foci of damage induced by cisplatin and especially by cisplatin/gentamicin in all glomerular components: glomerular capillaries, including their endothelial cells; basement membrane, epithelial podocytes, mesangial cells, and parietal cells of Bowman's capsule. The damage was expressed by endothelial cytoplasmic extrusions into the vascular lumen, thickening and lamination of capillary basement membrane, focal foot process fusion of podocytes, vacuolization in cytoplasm of endothelial cells of epithelial podocytes and of parietal cells, and the presence of lipid bodies and myeloid bodies in all glomerular cell types. Additionally, injurious effects to cytoplasmic organelles such as mitochondria, nuclei, and endoplasmic reticulum were observed. The results indicate that cisplatin alone and in combination with gentamicin is toxic to renal glomerular tissue. Since these drugs were previously found toxic for strial capillaries in the inner ear and since the main glomerular component is the glomerular capillaries, potential vascular damage and vascular complications in different body systems have to be taken into consideration when these drugs are needed in clinical use. PMID- 12537763 TI - The protective effect of ZnCl2 pretreatment on the development of postanoxic neuronal damage in organotypic rat hippocampal cultures. AB - Zinc is one of the trace elements playing an important role in many fundamental biological processes. However, it is also one of the possible etiological agents involved in nerve cell damage in certain human neurodegenerative disorders. The precise mechanism of neuroprotective ability of Zn against neurotoxicity evidenced in various pathological conditions remains unclear, especially concerning the intrinsic potential toxicity of this metal. This ultrastructural study was undertaken to determine the effect of zinc on the evolution of anoxia induced neuronal injury in the organotypic cultures of rat hippocampus. The in vitro model of oxygen deprivation was produced by maintaining the cultures in a pure nitrogen atmosphere in flask adapted for permanent gas flow for 20 min. The selected cultures were pretreated with micromolar concentration of ZnCl2 (25-500 microM) at 30 min prior anoxia. The ultrastructural findings documented that Zn exhibited dose-dependent ability to reduce anoxia-induced neuronal changes in hippocampal neurons in vitro. Zn at a concentration of 100 microM was able to significantly protect the hippocampal formation against the development of late apoptotic changes, whereas the early necrotic anoxia-induced neuronal injury was not so efficiently reduced. PMID- 12537764 TI - Endoneurial microangiopathy of sural nerve in experimental vacor-induced diabetes. AB - The pathogenic mechanism of diabetic peripheral neuropathy is multifactorial. This study investigated microangiopathic changes of endoneurial vessels of sural nerve in Vacor-induced diabetic rats. Experimental rats were divided into 3 groups: acute (80 mg/kg, once), chronic diabetic (8 mg/kg, 14 times), and control. Ultrastructural morphometric parameters of endoneurial microvessels, blood glucose, and body weight changes were statistically analyzed, and correlations among the variables were evaluated. The body weight and blood glucose levels were significantly different between the control and diabetic rats. The blood glucose was more significantly elevated in the acute diabetic rats than the chronic diabetic rats. Inner and outer vascular circumference, vascular area, luminal area, basement membrane area, and basement membrane thickness were significantly increased in the acute diabetic rats compared to the control group, but not between the chronic diabetic rats and the control group. The thickness of basement membrane was positively correlated with hyperglycemia and body weight loss (P < .01). These results suggest that the microangiopathy of the peripheral nerve could be developed in acute Vacor-induced diabetes. The decreased perfusion through microangiopathic endoneurial vessels appears to have a pathogenic role causing diabetic peripheral neuropathy. PMID- 12537766 TI - Invasive lobular carcinoma in a hypoplastic breast. AB - Breast hypoplasia is encountered as part of genetic syndromes or as a result of iatrogenic factors. The incidence of this malformation and the occurrence of breast carcinoma in such cases are unknown. The authors present a 66-year-old patient with a severe breast hypoplasia and invasive lobular carcinoma. The advanced clinical stage required neoadjuvant chemotherapy. After 5 CMF cycles with no significant effect, a modified radical mastectomy with axillary lymph node dissection was performed. The pathological report revealed an infiltrating lobular carcinoma with combined classical and alveolar growth and with minor morphological changes after the chemotherapy. Immunostaining for cell proliferation markers, apoptotic regulators, and cell adhesion molecules, such as the CD44 family and members of the cadherin-catenin group, was performed. The tumor expressed a high bcl-2/low bax ratio and lacked p53 immunoreactivity, which could explain the resistance to neoajuvant therapy. The lack of adhesion molecules, except for strong E-cadherin and beta-catenin reactivity, and weak CD44v6 expression were demonstrated. To the authors' knowledge this is the first case of an invasive lobular carcinoma in a hypoplastic breast reported in the English literature. PMID- 12537765 TI - Ultrastructurally "invasive" microvilli in an aggressively metastasizing biphasic malignant mesothelioma. AB - The presence of long, slender, often branching microvilli on cell surfaces is a characteristic feature of malignant and benign mesothelial cells. However, these typical microvilli are seen only in better-differentiated lumens within epithelial areas of malignant mesotheliomas. Presented here are the clinical and ultrastructural findings in a biphasic malignant mesothelioma that lacked lumens, but possessed very long microvilli. These invaginated deeply into the cytoplasm of neighboring tumor cells, as well as into the surrounding stromal matrix. The tumor cells had well-formed intercellular desmosomal junctions. The primary tumor was localized to the pleura and invaded the chest wall, but only minimally the lung. Lobectomy demonstrated the presence of metastatic tumor in 2 peribronchial lymph nodes. The disease progressed rapidly and, within 6 months, killed the patient. An autopsy revealed widespread metastases in multiple systemic organs. The authors speculate that the unique ultrastructural features of this case may be a clue to the unusually aggressive course of the neoplasm. PMID- 12537767 TI - Ribosome-lamella complex of leukemic cells in a patient with aggressive NK cell leukemia. AB - A 36-year-old male was admitted to the hospital with anemia and leukopenia. A bone marrow specimen revealed the proliferation of leukemic cells with flower like nuclei observed in adult T-cell leukemia. The leukemic cells were positive for HLA-DR, CD2, CD7, and CD56. A diagnosis of aggressive NK cell leukemia was made, the patient was treated with induction chemotherapy and cord blood stem cell transplantation, and he is well now. Seen with electron microscopy, some leukemic cells had ribosome-lamella complexes (RLC). This is the first reported case of leukemic cells with flower-like nuclei seen with light microscopy and RLC seen with electron microscopy. PMID- 12537768 TI - Follicular bronchiolitis (FBB) associated with Legionella pneumophilia infection. AB - An 8-month-old girl with respiratory distress and stridor was admitted to the authors' hospital. Two days later, she died of respiratory insufficiency due to pneumonia. Autopsy confirmed the presence of follicular bronchiolitis (FBB) in both lungs. After considering her clinical course, the authors focused on three pathogens: Legionella pneumophilia, Pneumocystis carinii, and Mycobacterium tuberculosis. Only Legionella pneumophilia was detected by both immunohistochemistry and PCR. PMID- 12537769 TI - Molecular analysis and pathology of a second pediatric HIV-associated Burkitt lymphoma. AB - Diagnosis of a second HIV-associated non-Hodgkin lymphoma (HIV-NHL) is rare, but additional cases may occur as aggressive therapy for both HIV and NHL improves. An 11-year-old presented with a second primary HIV-NHL following remission for 9 years. Analysis of the tumor demonstrated presence of EBV and HIV with absence of CMV, HHV-8, and HHV-6. Although microscopic disease was present only in CSF, analysis of peripheral blood and bone marrow by PCR was positive. The patient underwent a stem cell transplant, but within 3 months, his disease recurred. Analysis for residual disease and viruses in similar cases may provide information in understanding pediatric HIV-NHL. PMID- 12537770 TI - Expression and regulation of the transcriptional repressor ZNF43 in Ewing sarcoma cells. AB - In vitro, cells derived from Ewing sarcoma (ES) with the characteristic somatic rearrangement between the genes EWS and FLII can be induced to differentiate toward a neuronal phenotype by exposure to agents such as dibutyryl cyclic AMP (db cAMP) or retinoic acid. Therefore, expression of the chimeric Ews-Flil protein does not irreversibly block the capacity of Ewing cells to engage in the neuronal differentiation program initiated by these agents. To identify genes that might be involved in the maintenance of Ewing cells in their undifferentiated state, a PCR-based differential display method was used to compare gene expression patterns in Ewing cell lines with those induced to differentiate toward a neuronal phenotype. A cDNA was expressed at high levels in proliferating Ewing-derived EW-1 cells and downregulated in EW-1 cells induced to differentiate, which corresponds to ZNF43, a multi-zinc finger protein containing the Kruppel-associated box (KRAB) transcriptional repression domain. Treatment of EW-1 cells with antisense oligonucleotides complementary to ZNF43 mRNA induces morphological differentiation and growth arrest. These findings suggest a role for ZNF43 in the maintenance of ES cells in an undifferentiated state, and that ZNF43 could be a primary target for differentiation stimuli in Ewing cells. PMID- 12537771 TI - Congenital heart disease and its association with other congenital malformations found at autopsy. AB - The objective was to determine if a study of other malformations found autopsy in patients with congenital heart disease would contribute to an understanding of the mechanisms involved in the formation of these anomalies. In a large general hospital autopsies in children with congenital heart disease were selected, and the different cardiopathies were divided into 3 groups: those with isolated heart lesions, those with single gene mutants, as well as chromosome malformations, and those with idiopathic malformations not associated with a genetic syndrome. Because the cardiopathies most often associated with genetic syndromes were the septal defects in general (p = .001), it was presumed that these are influenced to a considerable extent by genetic factors. The association of left heart hypoplasia and coarctation of the aorta with multiple idiopathic malformations, particularly in the lower half of the body (p = .002), suggests that the latter may be due to vascular disruptions, because of the interruption of the flow of oxygenated blood in the embryo-fetus produced by these heart defects. Cranial defects were not associated with left heart obstruction, and are therefore unlikely to be produced by vascular disruption. PMID- 12537772 TI - Chorangiopagus parasiticus twins. AB - Chorangiopagus parasiticus twins (CAPP) occur due to asymmetric abnormality of monozygous, twinning. It is the commonest form of asymmetric twinning, occurring in 1% of monozygous twins. The asymmetric twin survives by parasitizing the more normally developed co-twin by connecting with the chorionic circulation, hence, chorangiopagus parasiticus. CAPP twin is a markedly edematous foetus with relatively well-developed legs, incomplete pelvis and lower spine. The body cavity contains some incomplete abdominal viscera but usually no thoracic organs. The upper portion of the twin consists of edematous cystic tissue. This parasitic perfused twin has no placental vascular connection and its cord vessels are conjoint with those of the parasitized pump twin on the surface of the placenta or somewhere along the cord. This malformation can be identified by ultrasound as early as 12 weeks gestation. PMID- 12537773 TI - Reduced acute phase response to differentiate between viral and bacterial infections in children. AB - The objective of this study was to document the reduced acute phase response that appears in children with viral as opposed to bacterial infections. The white blood cell count (WBCC), the erythrocyte sedimentation rate (ESR), and leukocyte and erythrocyte adhesiveness/aggregation were determined in 36 children with acute bacterial infection, 29 children with viral infection, and 19 controls. A significant reduced WBCC, ESR, and leukocyte and erythrocyte adhesiveness/aggregation was noted in the children with acute viral infection as opposed to those with bacterial infection: 10,800 +/- 3600 and 20,000 +/- 10,000 cells/cm2, 29 +/- 21 and 53 +/- 35 mm Hg, 23 +/- 9 and 41 +/- 15%, and 3.4 +/- 5.1 and 9.8 +/- 13.6 microns, respectively. The results indicate that a reduced acute phase response can be observed in children with an acute viral infection. This can have diagnostic implications and pathophysiological consequences in terms of less flow impairment in the microcirculation due to less red and white blood cell aggregation. PMID- 12537774 TI - Gonadotrophin-releasing hormone (GnRH) and GnRH agonists: mechanisms of action. AB - The hypothalamic decapeptide gonadotrophin-releasing hormone (GnRH) binds to specific receptors on pituitary gonadotrophs. These receptors belong to the family of G protein-coupled receptors. Their activation leads to phosphoinositide breakdown with generation of inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) and diacylglycerol. These second messengers initiate Ca2+ release from intracellular stores and activation of protein kinase C, both of which are important for gonadotrophin secretion and synthesis. Prolonged activation of GnRH receptors by GnRH leads to desensitization and consequently to suppressed gonadotrophin secretion. This is the primary mechanism of action of agonistic GnRH analogues. By contrast, GnRH antagonists compete with GnRH for receptors on gonadotroph cell membranes, inhibit GnRH-induced signal transduction and consequently gonadotrophin secretion. These compounds are free of agonistic actions, which might be beneficial in certain clinical applications. PMID- 12537776 TI - Direct ovarian effects and safety aspects of GnRH agonists and antagonists. AB - The wide application of gonadotrophin-releasing hormone (GnRH) analogues in assisted reproduction has raised concerns about their potential extra-pituitary actions. Although data from animal studies support the presence of ovarian GnRH receptors and of direct ovarian effects of GnRH analogues, this has not been unequivocally demonstrated in humans. Available evidence suggests, however, the existence of GnRH receptors in human granulosa-luteal cells, while the majority of the studies performed showed that progesterone is inhibited by GnRH agonists in vitro. On the other hand, no difference seems to exist between agonists and antagonists in terms of granulosa-luteal cell steroidogenesis under basal conditions. So far, no studies have suggested an adverse effect of GnRH analogues on the developing human follicle. Moreover, the physiological significance of a direct in-vitro ovarian effect of GnRH analogues remains to be established. PMID- 12537777 TI - The role of GnRH antagonists in assisted reproduction. AB - The different schemes of application of gonadotrophin-releasing hormone (GnRH) antagonists in ovarian stimulation protocols for assisted reproduction are reviewed. Starting from the initial schemes of single and/or multiple doses, the efficacy of GnRH antagonists in 'soft' protocols of ovarian stimulation has been evaluated in natural cycles, and the possibility of inducing the final oocyte maturation under antagonists either with native luteinizing hormone-releasing hormone or with GnRH agonists. The largest clinical studies carried out with cetrorelix or with ganirelix and published to date were analysed. Apart from the benefits of reducing the duration of the treatment, gonadotrophin requirements and ovarian hyperstimulation syndrome incidence, there is still a serious debate on the potential deleterious effect of GnRH antagonists on implantation and pregnancy rates. More specific aspects such as cycle programming, application of GnRH antagonists in protocols for poor responders etc., were also reviewed in this survey. PMID- 12537778 TI - GnRH antagonists: do they open new pathways to safer treatment in assisted reproductive techniques? AB - GnRH antagonists (GnRHnt) induce a rapid decrease in LH and FSH, preventing and interrupting LH surges. Their properties do not require a desensitization period and allow their use in the late follicular phase. GnRHnt could replace GnRH agonists (GnRHa) in ovarian stimulation without their side-effects and their long period of desensitivity. Both protocols using GnRHnt were associated with a smaller amount of gonadotrophin, a shorter stimulation period and a lower incidence of ovarian hyperstimulation syndrome (OHSS) with statistically comparable pregnancy rates. There is room for improvement of both protocols concerning the programming, the antagonist dose and the timing of antagonist administration. Luteal support should be maintained until the results of further studies. The perinatal outcome appears to be similar to that of other stimulation regimens. Triggering of ovulation can be facilitated by GnRHa for patients at risk of OHSS. Provided that pregnancy rates equivalent to those obtained with GnRHa are confirmed in larger series, GnRHnt will clearly represent a safer alternative to GnRHa protocols in IVF. PMID- 12537779 TI - The luteal phase after ovarian stimulation. AB - Evaluation of the luteal phase after ovarian stimulation presents several difficulties. Until today, it has not been proved that in cycles stimulated with clomiphene citrate/human menopausal gonadotrophins, luteal supplementation with progesterone significantly increases implantation rate. On the contrary, it is accepted that in cycles stimulated with GnRH agonists/gonadotrophins, support of the luteal phase is essential for the achievement of pregnancy. In GnRH antagonist cycles luteal supplementation, although widely practised, is not at present supported by randomized controlled trials. Finally, it appears that the combination of gonadotrophin stimulation with human chorionic gonadotrophin results in an inadequate luteal phase with or without the use of either agonists or antagonists. PMID- 12537780 TI - Ovarian stimulation for IVF and endometrial receptivity--the missing link. AB - The contemporary approach to ovarian stimulation for IVF treatment results in supraphysiological concentrations of steroids during the follicular and luteal phases of the menstrual cycle. These sex steroids act directly and indirectly to mature the endometrium, influencing receptivity for implantation. Corpus luteum function is distinctly abnormal in IVF cycles, and therefore luteal support is widely used. Various reasons may underlie the defective luteal phase, including (i) ovarian hyperstimulation per se, (ii) gonadotrophin-releasing hormone (GnRH) analogue co-treatment and (iii) the use of human chorionic gonadotrophin (HCG) to induce final oocyte maturation. The recent introduction of GnRH antagonist co treatment for the prevention of a premature LH rise during the late follicular phase allows for different approaches to ovarian stimulation for IVF. However, a recent meta-analysis showed that implantation rates may be compromised by using GnRH antagonists in currently employed regimens. The development of endometrium receptive to embryo implantation is a complex process and may be altered by inappropriate exposure to sex steroids in terms of timing, duration and magnitude. New approaches to the assessment of endometrial receptivity are now required. Novel approaches to ovarian stimulation aimed at adjusted GnRH antagonist regimens and achieving a more physiological luteal phase endocrinology are now appearing in the literature and may represent an important step in the improvement of the overall health economics of IVF. PMID- 12537781 TI - Molecular basis of implantation. AB - The process of embryonic implantation needs coordinated development and communication between the blastocyst and the maternal endometrium. Considerable advances in the understanding of the cell biology of the human embryo and maternal endometrium have been achieved separately. However, the communication between them, and the reciprocal effects on each other, constitute a complicated and yet unsolved question in reproductive medicine that must be further investigated. A paracrine dialogue between the embryo, endometrium and the corpus luteum is known to occur in other species such as rodents and primates. The aim of this work is to present updated information on the embryonic regulation of endometrial epithelial molecules such as adhesion molecules, anti-adhesion molecules, cytoskeletal proteins, chemokines and leptin during the apposition and adhesion phases of human implantation. PMID- 12537782 TI - Outcome of cryopreserved pronuclear oocytes obtained after ovarian stimulation with either HMG or recFSH and the GnRH-antagonist cetrorelix. AB - The present retrospective study evaluated the outcome of frozen-thaw cycles with oocytes obtained either during a multiple dose protocol of cetrorelix, or after the use of a gonadotrophin-releasing hormone (GnRH) agonist. A total of 101 subfertile couples were included. These couples had a total of 222 transfers of frozen-thawed pronuclear oocytes after IVF/intracytoplasmic sperm injection (ICSI) treatment. According to the stimulation protocol during various cycles, four groups were established: cetrorelix/recombinant FSH (recFSH) (69 cycles), cetrorelix/human menopausal gonadotrophin (HMG) (10 cycles), GnRH-agonist/recFSH (71 cycles) and GnRH-agonist/HMG (72 cycles). The transfer cycles were mildly stimulated with transdermal oestradiol. No statistically significant difference was seen among the four groups regarding post-thaw survival rate, cumulative embryo score, implantation rate and pregnancies. Frozen-thawed pronuclear oocytes obtained with the use of cetrorelix give satisfactory implantation and pregnancy rates, similar to those obtained with a GnRH-agonist. These results do not depend on the gonadotrophins (HMG or recFSH) used in the collecting cycle. PMID- 12537783 TI - The need for new methods of ovarian stimulation. AB - It is easy to diagnose sterility but much harder to diagnose subfertility, because minor defects are not necessarily associated with consistent impairment of fertility. Correct evaluation of the background fertility can avoid needless treatment of couples. Ovarian stimulation is a powerful strategy frequently used to re-establish or improve fertility in infertile couples, but it is often the cause of twin pregnancy. The risks and costs of multiple gestation are important factors that must be taken into full account in the pro-fertility decision. To avoid twins, inseminations should preferably be done in unstimulated cycles or in association with induced mono-ovulatory cycles. For the same reason, it is probably time to follow the policy of favouring single embryo transfer in young women undergoing IVF and intracytoplasmic sperm injection programmes. In addition, there is an urgent need to avoid an invasive multiple transfer policy in women over 38 and to select new strategies to improve the well-known age related lower implantation rate. PMID- 12537784 TI - Safety aspects of gonadotrophin-releasing hormone antagonists in ovarian stimulation procedures: ovarian hyperstimulation syndrome and health of children born. AB - The safety of ovarian stimulation procedures or the procedure of assisted reproduction in general can be estimated by various parameters. Two of the most important are the health of children born after the procedure and the incidence of ovarian hyperstimulation syndrome (OHSS). The latter is important because it is the most severe, potentially life-threatening complication of any stimulation procedure. The use of gonadotrophin-releasing hormone (GnRH) antagonists in ovarian stimulation protocols has had no impact on the health of children born. This was proven in 227 children born after the use of cetrorelix and in 73 children born after the use of ganirelix. To analyse the incidence of OHSS and the impact of GnRH antagonists on clinical pregnancy rates compared with the long protocol, a meta-analysis was done. This showed a reduction of OHSS with the use of cetrorelix. Furthermore, when compared with the long protocol, clinical and ongoing pregnancy rates were not significantly reduced with the use of cetrorelix. Taken together, the use of GnRH antagonists are safe with regard to children's health. The incidence of OHSS does not increase with ganirelix, and a reduction can be expected with cetrorelix. PMID- 12537785 TI - Will GnRH antagonists assist in the treatment of benign gynaecological diseases? AB - While GnRH agonists have become well-established tools for preoperative treatment of uterine fibroids or postoperative treatment in endometriosis for 3-6 months, GnRH antagonists seem to offer important advantages due to their specific pharmacological mode of action. Avoiding any flare-up effect, it seems to be possible to reduce treatment time to about only 2-4 weeks in the case of fibroids to obtain a clinically relevant reduction in size. Furthermore, due to the classic competitive receptor blockade induced by GnRH antagonists, it is feasible to preserve residual oestradiol secretion for a period of 8 weeks in patients with endometriosis. Endometriosis patients undergoing this treatment reported a symptom-free period, with no signs of mood changes, hot flushes, loss of libido, vaginal dryness or other symptoms. Serum oestradiol oscillated around a mean level of 50 pg/ml during therapy. Sequential administration of the GnRH antagonist cetrorelix (Cetrotide), in a 3 mg dosage once weekly over 8 weeks in the case of endometriosis or administration every 4th day for a time span of 2-4 weeks for fibroids, creates a new opportunity for medical treatment. Although 3 mg of cetrorelix acetate obviously acts as an intermediate depot preparation, results obtained so far are very preliminary. PMID- 12537786 TI - Ovarian stimulation: from basic science to clinical application. AB - Treatment for infertility, including ovarian stimulation, was first introduced almost 100 years ago. At this time, radiation therapy became an established treatment, and it was only some decades later that the problem of radiation induced cancer emerged. Non-human gonadotrophins, such as pregnant mare serum gonadotrophin (PMSG), and human pituitary gonadotrophins (HPG), were commonly used for hormonal stimulation procedures. However, use of PMSG led to antibody formation, and it was therefore only useful for the first treatment cycle. HPG produced good results, but its use came to an end in the late 1980s when it was linked to the development of Creutzfeldt-Jakob disease. The first hormonal product from human menopausal urine to be used was human menopausal gonadotrophin (HMG), followed later by purified preparations of this product. All of these preparations contained a high percentage of unknown urinary proteins, which interfered with batch-to-batch consistency. This changed with the introduction of recombinant gonadotrophins, produced from an immortalized/standardized mammalian cell line (CHO). More recent developments include the introduction of long-acting gonadotrophin formulations. The development of gonadotrophin-releasing hormone (GnRH) analogues and more recently the use of GnRH antagonists has helped to improve ovarian stimulation protocols by optimizing their efficacy, and making them easier to administer. PMID- 12537787 TI - "Legal cases spell big trouble and great opportunities for IVF embryologists": a comment from the perspective of the HFEA. PMID- 12537789 TI - Will costs of ICSI be reimbursed for patients in Germany? PMID- 12537788 TI - Personal indemnity for professionals in reproductive medicine. PMID- 12537790 TI - Altruism in assisted reproductive technologies. AB - What makes reproductive technology unique is that it is in the forefront of creating human life, with an emphasis on 'giving'. Not surprisingly therefore, a significant appeal to altruism has crept into the language of reproductive technologies. If altruistic donations and behaviour involving reproductive technologies are to be encouraged we need to have a uniform understanding of the terminology. For this we need to explore how the term altruism has been used historically and is being used in modern language. It is also important to acknowledge that gender differences may influence altruistic behaviour. The suggested working definition for altruism has three major components; a motivational state, an ultimate goal and a desire to increase another's welfare. Within this framework definition, self-benefit is permissible provided the ultimate goal is not self-benefit. Altruistic donation is possible provided receiving money is not the primary motive. It is important in our efforts to improve the quality of donor and donations, which are central to assisted reproductive technologies, that any reference to altruism be restricted to the situation where only the ultimate goal, with a single motive, can be discerned. The success of recruiting altruistic donors will inevitably be associated with the attitudes and structures in which reproductive technologists operate. Through such transparency it is hoped more effective control will be achieved for donors over their donations. Irrespective of the setting, a true measure of an altruistic donation or behaviour can only be assessed by giving consideration to the primary motivational state, ultimate goal and benefit to another's welfare. PMID- 12537791 TI - Fertilizable oocytes reconstructed from patient's somatic cell nuclei and donor ooplasts. AB - The only assisted reproduction treatment now available for women with ovarian failure or irreparable oocyte defects is oocyte donation. However, some women experience psychological barriers to the recourse to donor oocytes, related to the lack of contribution of their proper genes to the progeny. A pilot study in humans suggests that this problem may be overcome by the development of techniques for haploidization of somatic cell nuclei, allowing the formation of new oocytes bearing the complete nuclear genome of the patient. Somatic cell nuclei were obtained from cumulus cells of a patient who failed to produce fertilizable oocytes and were transferred into enucleated oocytes (ooplasts) from a donor. Out of six ooplasts injected with the somatic cell nuclei and fertilized with spermatozoa from the patient's husband, signs of haploidization were detected in three oocytes, two of which subsequently started embryonic development and were cryopreserved for eventual future transfer to the genetic mother. These data show that human oocytes can be used for both reprogramming and haploidization of somatic cell nuclei, allowing reconstruction of genetically own oocytes for patients without, or with seriously disturbed, ovarian function. PMID- 12537792 TI - Placental transfer of temazepam and fentanyl in early human pregnancy. AB - The placental transfer of temazepam and fentanyl was studied in 37 women undergoing surgical termination of pregnancy between 12 and 17 weeks. The women received temazepam 10 mg, approximately 1 h pre-operatively and fentanyl 2 &mgr;g/kg intravenously, at induction of anaesthesia. A single 2 ml fetal blood sample was taken from each anaesthetized subject using an ultrasound-guided trans abdominal puncture at either 5, 10, 15 or 20 min after administration of fentanyl. A maternal blood sample was collected simultaneously and both were assayed for temazepam and fentanyl. Both drugs were detected in all fetal serum samples. Fetal:maternal (F:M) drug concentration ratio was 0.38 for temazepam, which did not change between 60 and 120 min after oral administration to the mother. Overall F:M ratio for fentanyl was 1.87 for fentanyl, which increased from 1.42 at 5 min to 2.6 at 20 min. F:M ratio of fentanyl concentrations increased with time (r = 0.48, P < 0.003), whereas F:M ratio of temazepam concentrations did not change significantly with time, between 50 and 120 min after the drug was administered to the mother (r = 0.32, P = 0.06). F:M ratios changed with gestational age of the fetus for both drugs, increasing as gestational age increased in the case of temazepam (r = 0.47, P = 0.003) and decreasing in the case of fentanyl (r = 0.38, P = 0.02). These changes can be explained by variation in uterine blood flow, trophoblast membrane thickness and feto-placental metabolism. These results also indicate that the in-vitro study of these drugs is unlikely to reflect the in-vivo situation in early pregnancy. PMID- 12537793 TI - Use of a non-contact, infrared laser for zona drilling of mouse embryos: assessment of immediate effects on blastomere viability. AB - To compare the effects of using a non-contact infrared laser with acid Tyrode's for zona drilling, mouse embryos were used to examine both immediate effects on blastomere viability, using fluorescent markers, and development to the blastocyst stage in vitro. Preliminary experiments with isolated blastomeres established that the safe working distance for the lowest setting of the laser (pulse duration 5 ms) was 8 micro m. At the 2-cell stage, when it was possible to drill through the zona while maintaining this safe distance from adjacent blastomeres, no immediate damage was detected. At the 8-cell stage, blastomeres are generally <8 micro m away from the outer surface of the zona and often touch its inner surface. Hence it was not possible to drill the zona while maintaining a safe distance and as a consequence almost all embryos had at least one, and often more, damaged blastomeres. Acid Tyrode's drilling also frequently lysed an adjacent blastomere at this stage and after 24 h in culture cavitation was delayed compared with laser-drilled embryos. Cell numbers at the late morula/early blastocyst stage, analysed by differential labelling of trophectoderm and inner cell mass nuclei, were significantly reduced following zona drilling. With the laser-drilled embryos, this was in proportion to the immediate effects on blastomere viability, whereas with acid Tyrode's, the reduction was greater, indicating an adverse effect on cleavage rate. It is concluded that use of a non-contact laser, at an appropriate pulse duration and distance from adjacent blastomeres, avoids blastomere lysis and adverse effects on development caused by acid Tyrode's zona-drilling. PMID- 12537794 TI - Possible ramifications of the identification of ovarian follicular granulosa cells as specialized endothelial-like cells: a speculative treatise. AB - This article presents several possible extensions and considerations stemming from the putative identification of cells of the ovarian follicular granulosa as specialized endothelial-like cells. The epithelial assignment for cells of the follicular granulosa is of limited value and potentially confusing. Phenotypic and biological evidence suggests some follicle cells may represent a 'primitive', or quasi-differentiated, cell population. Other evidence suggests that cells of the follicular granulosa produce a range of molecules whose synthesis may be affected by hypoxia; some of these molecules may play strategic roles during follicular maturation, ovulation and formation of the corpus luteum. Just as follicle cells appear capable of endothelial cell-like behaviour(s), endothelial cells appear capable of engaging in some activities essential to follicular function, including steroid hormone receptivity, hyaluronic acid synthesis and steroidogenesis. A consideration of the cells of the follicle as a specialized endothelial-like cell population may have important clinical applications with respect to the anomalies and pathologies associated with human infertility. In addition, the cells of the follicular granulosa may represent an unusual and potentially powerful model system with which to study and extend the scope of our current understanding of basic endothelial cell biology. PMID- 12537795 TI - New insights into maternal-fetal interactions at implantation. AB - The immunotrophic theory was enunciated by Tom Wegmann. Since then, the involvement of cytokines in implantation and materno-fetal tolerance has emerged as a central topic in reproductive immunology. This brief survey covers the historical background leading to the specification of the crucial role of cytokines at the feto-maternal interface, and the present known patterns of their function. Focus is addressed to the most recent concept, e.g. pregnancy as a Th2 phenomenon (the immune response of the mother is biased towards the production of anti inflammatory cytokines, amongst them IL-10 which suppress inflammatory responses). A brief description of the role of inflammatory cytokines in implantation is presented to explain why it does not fit into such a scheme, and other recent data, for example on gamma interferon, also fails to accord with the Th2 phenomenon. PMID- 12537796 TI - Menopause, a global perspective. AB - This paper attempts to provide a global perspective of the menopause as a period in a woman's life. It is growing in length, year after year and there is a need to place these years in the overall context of ageing. There are many fallacies about ageing. The most serious is the view describing old people exclusively as a burden for society, when in fact they can and should be considered as a resource. Women live longer almost everywhere in the world. Growing evidence indicates that the process of ageing differs in a considerable way in the two sexes. This difference may, at least in part, be due to sex-related differences in the regulation of stress response mediators. In addition, variability in the mitochondrial genome also displays a sex-specific impact on longevity. Restricting the discussion to longevity in the female, a paramount role must be given to sex hormones in improving a woman's ageing. Indeed, it is the fall in oestradiol production that qualitatively changes the ageing perspective in the human female, since oestrogen secretion plays a major role in guaranteeing a woman's psycho-physical equilibrium during the fertile period. These considerations represent the philosophical basis for post-menopausal hormone replacement therapy. PMID- 12537798 TI - Circadian pacemaking is shared between fibroblasts and suprachiasmatic nucleus. PMID- 12537797 TI - PGD protocols using multiplex fluorescent PCR. PMID- 12537800 TI - Internet and assisted human reproduction: internet, law and IVF. PMID- 12537799 TI - Internet and assisted human reproduction: reproductive services and the internet. PMID- 12537801 TI - The human embryo, embryonic stem cells, cloning, legal and political precepts. PMID- 12537802 TI - Preimplantation genetic diagnosis in Germany - ethical responsibility and law. AB - Preimplantation diagnosis (PGD) can be judged as early or brought forward prenatal diagnosis and brings up the ethical question in Germany whether potential selection of embryos and intentional killing of a human conceptus, showing a defect not or almost not compatible with life, is reconcilable with human dignity. Whereas prenatal diagnosis and its consequences are socially accepted in Germany, PGD is not compatible with the Embryo Protection Law and is still under debate since >5 years. On the other hand abortion is not subject to prosecution because the legal status of the mother outweighs that of the embryo or fetus. Here the conflict of two legally protected rights - the embryo's worth of protection and the woman's family planning - are accepted and form the basis of the potential decision of the prospective mother to carry the fetus to term or not. An obvious and deep contradiction of assessment is present between the legal requirements for the regulation of abortions in its amendment of 1995 and the Embryo Protection Law which exists unchanged since 1991. The killing of an embryo or fetus in vivo is not subject to prosecution although it is illegal, whereas the discard of an embryo in vitro after PGD was performed, also illegal, is subject to prosecution. PMID- 12537803 TI - The Indian Council of Medical Research issues a statement on assisted reproductive techniques. PMID- 12537804 TI - Analysis of intracytoplasmic sperm injection procedures related to delayed insemination and ejaculated, epididymal and testicular spermatozoa. AB - In all, 1210 treatment cycles were divided into three categories for retrospective analysis according to the period of delay between oocyte retrieval (occurring at a fixed time after human chorionic gonadotrophin) and intracytoplasmic sperm injection (ICSI) of <3 h, 3-5 h, >5 h (referred to as 'delayed ICSI'). Three stages from oocyte to the birth of a live baby were identified for statistical analysis, (i) fertilization (2PN zygotes), (ii) cleavage of 2PN zygotes, (iii) transferred embryo to live birth. Stages 1, 2 and 3 were analysed statistically for the three time periods. Chi-square analysis showed no significant effect of delayed ICSI on fertilization (chi(2) = 3.615, P = 0.65), and embryo transfer to birth (chi(2) = 1.840, P = 0.399). The effect on cleavage was significant (chi(2) = 9.625, P = 0.008). However, shorter incubation times produced results which were better than the traditional longer ones. The success rate at the cleavage stage was so high that the marginal advantage had very little effect on the overall process. This study of a substantial patient sample establishes that ICSI on a peri-ovulatory oocyte (<3 h after oocyte retrieval) does not compromise outcome parameters, and that longer periods of incubation (>5 h) do not offer a statistically significant advantage. PMID- 12537805 TI - Pregnancies from single normal embryo transfer in women older than 40 years. AB - The aim of this retrospective study was to determine the pregnancy rate from the transfer of single genetically normal embryos in patients of advanced reproductive age. The study group included 23 patients (mean age 42.2 +/- 1.3 years) who underwent 27 in-vitro fertilization (IVF) cycles in which preimplantation genetic diagnosis (PGD) was carried out on single blastomeres from day 3 embryos. The control group included 54 patients (mean age 43.3 +/-1.9 years) who underwent 69 cycles of IVF without PGD. Ovarian stimulation in all patients consisted of follicular phase leuprolide acetate administration, followed by ovulation induction with gonadotrophins. The mean number of biopsied embryos was 5.6 +/- 0.5. No embryo transfer occurred in six patients (10 cycles) because all embryos biopsied were abnormal. Seventeen patients (17 cycles) each had one genetically normal embryo transferred resulting in six on-going clinical pregnancies (35% per embryo transfer cycle). The mean number of embryos transferred in the control group was 4.0 +/- 0.8. Nineteen clinical pregnancies were obtained in 69 transfer cycles in the control group (28% per embryo transfer cycle). The transfer of a single normal embryo in patients of advanced reproductive age can lead to acceptable pregnancy rates. Aneuploidy appears to be a major cause of reproductive failure in this group of patients. PMID- 12537806 TI - Preimplantation diagnosis for p53 tumour suppressor gene mutations. AB - Preimplantation genetic diagnosis (PGD) was introduced for high-risk couples to avoid establishing affected pregnancies potentially requiring termination following prenatal diagnosis. This opens the possibility for PGD for late onset disorders with genetic predisposition, including inherited cancer predisposition, because only embryos free from the predisposing gene may be transferred back to the patient, with no potential risk for pregnancy termination. PGD was performed for two couples, one with maternally and one with paternally derived p53 tumour suppressor mutations, 902insC in exon 8 and G524A in exon 5, respectively. This involved a standard IVF protocol, allowing oocytes or embryos to be tested prior to their transfer back to uterus. Maternal mutation was tested by sequential PCR analysis of the first and second polar bodies, removed following maturation and fertilization of oocytes, while paternal mutation analysis required embryo biopsy at the cleavage stage. To avoid misdiagnosis due to allele drop out, multiplex nested PCR was applied, involving p53 mutation analysis simultaneously with the linked short tandem repeats in intron 1. Of 10 oocytes tested in two PGD cycles for 902insC mutation, four unaffected oocytes were pre-selected for transfer yielding no clinical pregnancy. Of 18 embryos analysed in two cycles for G524A mutation, seven mutation-free embryos were detected, two of which were transferred in each cycle, resulting in a singleton pregnancy and birth of a mutation-free child. This is the first PGD for inherited cancer predisposition determined by p53 tumour suppressor mutations, resulting in a clinical pregnancy and birth of a child free from inherited cancer predisposition. PMID- 12537807 TI - An introduction to reproduction in pharaonic Egypt. AB - Reproductive science is a new-born medical speciality compared with other specialities such as surgery or dentistry. However, infertility and obstetrical complications commonly diagnosed in the 20th century have a history as old as medicine itself. In an attempt to distance themselves from the African and Middle Eastern roots of our culture, the European intellectuals of the 18th and 19th century have written the history of the democratic world starting with the Greco Roman period. Egyptian medicine has influenced the medicine of neighbouring civilisations, including the culture of ancient Greece and its influence spread onward, thereby affecting Western civilisation. The aim of this review is to give credit to these early researchers, who often concluded with extremely accurate observations and to present their colourful and sometimes controversial contribution to our knowledge of reproductive anatomy, physiology and pathology. PMID- 12537808 TI - Growth of human preimplantation embryos in vitro. AB - The human oocyte fertilizes and develops into embryos in the Fallopian tube and reaches the uterus only after compaction. However, for several years embryos that were developed following in-vitro fertilization (IVF) were transferred into the uterus on day 2 or 3 at the 4-8 cell stage in contrast to the in-vivo situation where they would be present in the Fallopian tube. Earlier attempts to grow embryos in vitro for 5 to 6 days were not always successful. Attempts were therefore made to understand the in-vivo environment of the Fallopian tube where the early embryonic development occurs. This article reviews the studies carried out to understand the composition of fluids in the Fallopian tube specifically with reference to the energy metabolites - lactate, pyruvate and glucose; it also covers how the formulation of culture media for human IVF and embryonic development were modified over the years based on some classical work done on embryo culture in laboratory animals. PMID- 12537809 TI - The role of LHRH agonists and antagonists. AB - Administration of GnRH analogues (agonists as well as antagonists) produces suppression of the pituitary---gonadal axis, thus inhibiting the secretion of LH, FSH and sexual steroids. For this reason, analogs are indicated in all those clinical situations where suppression of gonadotrophins (precocious puberty, contraception) or of sexual steroids (endometriosis, prostate hyperplasia, cancer, uterine fibroids) is desired. For several years GnRH agonists have been used in combination with gonadotrophins for ovarian stimulation for assisted reproduction in order to control premature LH surges and to reduce cancellation rate with improvement of the pregnancy rate per cycle. This effect is obtained after 2 weeks of agonist administration. The immediate suppression of the pituitary achieved by GnRH antagonists without an initial stimulatory effect is the main advantage of these compounds over the agonists. The prevention of a premature LH surge by GnRH antagonists can be obtained by multiple dose or by a single administration. Both protocols offer the following advantages over the agonists: they require fewer ampoules of gonadotrophins, shorter duration of stimulation, there is a preserved pituitary response to GnRH, less risk of ovarian hyperstimulation syndrome and the luteal phase seems to be more preserved. The main disadvantages of the antagonists are that they are expensive and that pregnancy rate appears to be slightly lower than with the agonists. GnRH antagonists will probably replace agonists in ovarian stimulation treatment for assisted reproduction techniques. PMID- 12537810 TI - Results of assisted reproduction techniques in Latin America. AB - The fact that today it is possible to write a comprehensive report on assisted reproduction technologies in Latin America is the result of a serious and systematic effort, accomplished by more than 80 centres, from Mexico to Chile. Over the past 10 years, these centres representing the vast majority of assisted reproductive treatment cycles performed in Latin America, have agreed voluntarily to report their work to our regional registry using a pre-established format. Furthermore, during 1999 and part of 2000, all centres were visited by a team of biologists and clinicians, who evaluated for consistencies in the data reported. This activity also included an evaluation of personnel, facilities, etc. It can be proudly said that the data reported are a true reflection of the actual situation in our region. PMID- 12537811 TI - Genetics of human premature ovarian failure. PMID- 12537812 TI - Natural human dizygotic twinning: are mutations in the FSH receptor responsible? PMID- 12537814 TI - Wrongful life verdict in Paris. PMID- 12537813 TI - Stores of inexpensive stem cells promise lifelong health. PMID- 12537815 TI - Developing countries need drugs at affordable cost for prevention of AIDS. PMID- 12537816 TI - Designer babies - are they a reality yet? A reply. PMID- 12537817 TI - Adam Nash: legally speaking, a happy ending or slippery slope? AB - The birth of Adam Nash, following IVF and then preimplantation genetic diagnosis (PGD) on the resulting 15 embryos to find which would be a potential bone marrow match for his older sibling, suffering from Fanconi's anaemia, is the first reported case of genetic selection of an embryo to save the life of an existing person. The case has stirred debates worldwide over the appropriateness and implications of using the technique for this and related purposes. Legally, it is suggested that embryos are indeed entitled to special respect because of their potential for life, but certain principles must not be overlooked, and the Nash case was wholly within acceptable legal principles. The legal perspective offered here concludes: (i) while embryos are entitled to certain protections, the mere fact that they are extracorporeal raises the danger that the rights and protections assigned to them will be wrongly elevated over the legally protected procreation rights of the adults who create them; (ii) divorce litigation involving "custody" of embryos is not a direct parallel and legal analogies must be distinguished; (iii) the status of embryos must be carefully defined; and (iv) a national or international, multi-disciplinary body should be created to grapple with the developing issues and uses that are sure to follow. PMID- 12537818 TI - Cancer patients, gametes, gonadal tissue, and the UK legal status. AB - With advances in reproductive technologies, there are new opportunities for preservation of fertility potential for cancer patients receiving damaging treatment regimens. These include cryopreservation of gonadal tissue and maturing germ cells. These developments were not envisaged in the UK Human Fertilisation and Embryology Act 1990. Complex legal interpretations have followed in deciding which techniques come under statutory remit of the Human Fertilisation and Embryology Act 1990, and whether a licence is necessary to conduct such activities. The decisions have depended on the legal definition of the gamete and the fact that substituted consent within the Act 1990 is specifically disallowed. In our analyses we believe several areas require further explanation or improvement: the definition relating to the oocyte, its applicability to ovarian tissue, a pre-Tanner stage 2 patient whose immature spermatozoa may satisfy the definition of gamete, and the legal mechanism of substituting consent which may allow the unregulated use of frozen gonadal tissue or germ cells for procreation in future years. In a recent development it appears that gonadal tissue may come under a 'tissue specific body' and not the Human Fertilisation and Embryology Authority. It makes sense from the standpoint of patient welfare and the limited public and clinical resources, to place under one regulatory body all biological material where the ultimate aim is human procreation. PMID- 12537819 TI - Does pronuclear morphology and/or early cleavage rate predict embryo implantation potential? AB - A total of 340 patients referred for in-vitro fertilization was included in a retrospective, comparative study in which zygotes were studied regarding alignment and polarization of nucleolar precursor bodies (NPB) and also early cleavage in relation to implantation and pregnancy rates for the 680 transferred embryos. At assessment of the pronucleus 18-19 h after sperm injection, NPB were checked for alignment/polarization. Twenty-six hours after sperm insemination the zygotes were assessed for early cleavage. At embryo transfer the two embryos with the best morphological score, irrespective of polarization and early cleavage, were selected for transfer. The overall rate of positive HCG tests 17 days after embryo transfer was 42% and the implantation rate 23%. Fourteen percent of the patients received two embryos with polarized NPB, with a positive HCG test of 51%. Embryo transfer with early-cleaved embryos was carried out in 21% of the cycles, with a pregnancy rate of 45%. Embryos with polarized NPB and/or early cleavage were transferred in 34% of the cycles, with a pregnancy rate of 51%, compared with a pregnancy rate of 38% when none of the embryos fulfilled these criteria (P-value 0.02). In this study the pregnancy rate was significantly higher when one or two embryos were polarized and/or early cleaved. It is concluded that in a cohort of morphologically good embryos, assessment for alignment/polarization of NPB and/or early cleavage can, together with conventional morphological criteria, serve as a simple non-invasive method for selection of embryos with high implantation potential. PMID- 12537820 TI - Preimplantation diagnosis for long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency. AB - Long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency is a fatal autosomal recessive metabolic disorder, presenting during infancy. Preimplantation genetic diagnosis (PGD) provides an option for establishing an unaffected pregnancy, avoiding the risk for termination of pregnancy following prenatal diagnosis. The method for pre-selection of mutation-free oocytes for LCHAD deficiency was developed by testing the first and second polar body removed from oocytes by micromanipulation techniques in the framework of in-vitro fertilization. To avoid misdiagnosis, testing was done using hemi-nested polymerase chain reaction (PCR), with outer primers designed to lie outside the pseudogene, eliminating false priming. Four of 12 tested oocytes were predicted to be unaffected, based on the heterozygous first and mutant second polar body. The embryos resulting from these mutation-free oocytes were replaced, yielding a singleton clinical pregnancy and birth of a healthy child following confirmation by prenatal diagnosis. PMID- 12537821 TI - Methods in preimplantation genetic diagnosis. AB - Preimplantation genetic diagnosis (PGD) is a new strategy, orientated toward primary prevention of congenital anomalies in couples with reproductive risk, such as advanced maternal age, carriers of chromosomal abnormalities, and carriers of monogenic conditions. For these patients, PGD is an acceptable alternative to prenatal diagnosis, mainly in those countries where pregnancy interruption is forbidden by law. PGD effectively avoids the implications linked to traditional prenatal diagnosis. Centres that provide medical servicies on reproductive biomedicine are responsible for the development and improvement of this new prevention strategy. Thanks to advances in micromanipulation techniques, associated with recent progress in molecular genetics, PGD may be employed for any genetic condition in the future. PMID- 12537822 TI - Should human embryos be transferred as cleaving embryos on day 3 or as blastocysts on day 5: the debate heats up! PMID- 12537823 TI - Immunocontraceptive potential of recombinant bonnet monkey (Macaca radiata) zona pellucida glycoprotein-C expressed in Escherichia coli and its corresponding synthetic peptide. AB - Zona pellucida (ZP) glycoproteins have been proposed as candidate antigens for development of immunocontraceptive vaccines. In this study, the efficacy to block fertility by immunization with recombinant bonnet monkey (Macaca radiata) zona pellucida glycoprotein-C (r-bmZPC) expressed in Escherichia coli and its synthetic peptide (P(4): KGDCGTPSHSRRQPHVVSQWSRSA, aa residues 324-347) conjugated to diphtheria toxoid (DT) has been evaluated in a homologous system. Female bonnet monkeys, immunized with P(4)-DT conjugate showed better immunocontraceptive potential as compared to an r-bmZPC-DT immunized group. In spite of high anti-P(4) antibody titres, animals continued to have ovulatory cycles and showed no disturbance in cyclicity (except summer amenorrhoea). No ovarian pathology was observed in the P(4) immunized group. These results suggest that immunization with the P(4) may lead to block in fertility without obvious ovarian dysfunction. However, further inputs are required to identify additional ZP based B-cell epitopes to enhance the contraceptive efficacy. PMID- 12537824 TI - Definition and causes of infertility. AB - Infertility is a common problem affecting one couple in six. It can be defined as the incapacity to fulfill pregnancy after reasonable time of sexual intercourse with no contraceptive measures taken. The evidence for changes in the prevalence of infertility is difficult to establish. This increase could be due to at least four factors: delayed childbearing, alterations in semen quality due to habits such as cigarette smoking and alcohol, changes in sexual behaviour and eliminations of most taboos. The study of infertile couple has always been focussed on different factors: ovulatory factor (present in about 20% of couples), utero-tubal peritoneal factor (present in ~30% of couples), semen migration factor (10% of cases) and male factor (30% of couples). Around 40% of all infertile couples exhibit a combination of factors and about 15% of couples may not display any objective alteration leading to a definite diagnosis. During the past two decades there have been three important changes in infertility practice. First, the introduction of assisted reproduction technologies has provided an opportunity to study basic reproductive processes. Second, societal changes have occurred such as the increase in the proportion of women over 35 years old seeking pregnancy. This fact is due to a later age for marriage and postponement of pregnancy. Third, the development of molecular biology and genetics has become very important for the study, diagnosis and assessment of couples, many of them considered until now as "unexplained infertile couples". PMID- 12537825 TI - Recombinant human follicle-stimulating hormone: a scientific step to clinical improvement. AB - Urinary-derived follicle-stimulating hormone (FSH) preparations have been used clinically for many years. Although effective, these have a number of disadvantages, not least of which is their variable composition. The availability of recombinant human FSH (r-hFSH), produced from CHO cells, with its constant composition and exceptionally high purity, has, therefore, aroused great interest. This review focuses on the use of r-hFSH for ovarian stimulation in assisted reproduction technology protocols and the treatment of World Health Organization Group I and II anovulation. The use of r-hFSH has been shown to lead to improvements in efficacy over urinary-derived preparations, particularly in assisted reproductive treatment, and a recent meta-analysis has shown higher ongoing pregnancy rates with the recombinant product. Although the two available recombinant products from CHO cells (follitropin alpha [Gonal-F((R))] and beta [Puregon((R))]) are similar from a physicochemical perspective, some minor advantages have been reported for follitropin alpha in relation to pregnancy rates and better local tolerance to injections. The apparent higher bioactivity of r-hFSH has led to reduced total FSH consumption over shorter treatment periods compared with conventional preparations, thus reducing overall exposure for patients. This is likely to confer not only safety benefits, but also cost effectiveness as demonstrated through pharmaco-economic modelling. PMID- 12537826 TI - Legal cases spell big trouble and great opportunities for IVF embryologists. PMID- 12537828 TI - Significant maternal effect on inheritance in 2-cell mouse embryos. PMID- 12537829 TI - Summaries for Students, Patients and General Readers. PMID- 12537827 TI - An unusual apoptotic protein active in germline and inner cell mass in mice. PMID- 12537831 TI - A cross-sectional study of occupational noise exposure and blood pressure in steelworkers. PMID- 12537830 TI - The effect on earmuff attenuation of other safety gear worn in combination. PMID- 12537832 TI - Aircraft noise exposure from schiphol airport: a relation with complainants. PMID- 12537834 TI - Excretion of Cortisol under Nocturnal Noise and Differences due to Analytic Techniques. PMID- 12537833 TI - Stress Hormone Changes in Persons exposed to Simulated Night Noise. PMID- 12537835 TI - Estimated leisure-time noise exposure and hearing symptoms in a finnish urban adult population. PMID- 12537836 TI - The Noise/Stress Concept, Risk Assessment and Research Needs. AB - In principle, the noise/stress hypothesis is well understood: Noise activates the pituitary-adrenal-cortical axis and the sympathetic-adrenal-medullary axis. Changes in stress hormones including epinephrine, norepinephrine and cortisol are frequently found in acute and chronic noise experiments. The catecholamines and steroid hormones affect the organism's metabolism. Cardiovascular disorders are especially in focus for epidemiological studies on adverse noise effects. However, not all biologically notifiable effects are of clinical relevance. The relative importance and significance of health outcomes to be assessed in epidemiological noise studies follow a hierarchical order, i.e. changes in physiological stress indicators, increase in biological risk factors, increase of the prevalence or incidence of diseases, premature death. Decision-making and risk management rely on quantitative risk assessment. Epidemiological methods are the primary tool for providing the necessary information. However, the statistical evidence of findings from individual studies is often weak. Magnitude of effect, dose-response relationship, biological plausibility and consistency of findings among studies are issues of epidemiological reasoning. Noise policy largely depends on considerations about cost-effectiveness, which may vary between populations. Limit or guideline values have to be set within the range between social and physical well-being - between nuisance and health. The cardiovascular risk is a key-outcome in non-auditory noise effects' research because of the high prevalence of related diseases in our communities. Specific studies regarding critical groups, different noise-sources, day/evening/night comparisons, coping styles and other effect-modifying factors, and the role of annoyance as a mediator of effect are issues for future research in this field. PMID- 12537837 TI - Chronic Cortisol Increases in the First Half of the Night Caused by Road Traffic Noise. AB - 56 children age 7 - 10 had a medical check-up and they and their mothers completed questionnaires. Additionally the children's excretion of free cortisol was measured by HPLC in two urine samples collected at 1 p.m. and in the morning. The children lived either at a busy road with 24 h lorry traffic or in quiet areas. At the side of the road the noise level was registered during five nights. In the bedrooms representative measurements of the short-term maximal sound level (L(Amax) and L(Cmax)) and of the frequency spectrum were taken. During the night on average every 2 minutes a lorry with L(max) > 80 dB(A) passed by the houses. The indoor levels of the higher exposed half of the children were L(max) = 33-52 dB(A) resp. 55-78dB(C)). The frequency spectrum had its maximum below 100 Hz. 74% of the higher exposed never opened their windows as compared to 25% in the lower exposed half group. The excretion of free cortisol and its metabolites in the first half of the night was significantly correlated to L(Cmax) (co-variables: age, sex, and the day of the week) as well as to impaired sleep, memory and ability to concentrate. The cortisol excretion in the second half of the night was not correlated to the noise level. Disturbances of the normal circadian rhythm of cortisol can be quantified by the quotient of the cortisol excretion in the first half of the night in relation to that in the second half. Children under long-term road traffic noise exposure during the night had an increased risk of chronic stress hormone regulation disturbances. These disturbances were significantly correlated to L(Cmax) and findings of allergy and/or asthma bronchial. Long-term low frequency noise exposure with Lmax < 55 dB(A) during the night resulted in chronic increases of children's excretion of free cortisol in the first half of the night and in serious disturbances of the circadian rhythm of cortisol. Indications of increased risks of asthma bronchial and allergies in noise exposed children with stress hormone regulation disturbances need further clarification PMID- 12537838 TI - The Joint Effect of Industrial Noise Exposure and Job Complexity on All-Cause Mortality - The CORDIS Study. AB - In a previous follow-up study of industrial workers (the CORDIS study, Melamed et al., 1999a) we demonstrated a dose-response relationship between occupational noise exposure levels and all-cause mortality. In that study the type of jobs that workers were engaged in was not taken into account. However, in further analyses of CORDIS data we have found that noise exposure is particularly detrimental to health for workers engaged in complex jobs. Therefore in this 12 year study we attempted to determine the combined effect of job complexity and noise exposure on all-cause mortality in 2606 industrial workers. We divided the workers into four groups based on a combination of either high or low noise exposure, and whether they performed simple or complex jobs. There was an increased risk for all-cause mortality (OR = 1.86, 95% CI = 1.04-3.32), in workers who performed complex jobs under high noise exposure levels compared to those who performed simple jobs under low noise exposure. This remained significant even after adjusting for possible confounding variables. There was a trend for a more pronounced effect among less educated workers, among blue-collar workers, and in those with higher tenure. We conclude that occupational noise exposure is associated with excess mortality risk among workers performing complex jobs. PMID- 12537839 TI - Prevalence of Arterial Hypertension and Myocardial Infarction in Relation to Subjective Ratings of Traffic Noise Exposure. AB - A cross-sectional study was performed on a 5% sample of the adult population of the city of Pancevo (3622 residents). The response rate was 79 % (2874), with 1243 interviewed males (43%) and 1631 females (57%). Noise annoyance was assessed on a five- grade verbal scale (Not at all; Slightly; Moderately; Very; Extremely). Arterial hypertension was defined by antihypertensive treatment, information on which was obtained from questionnaire. Myocardial infarction was also subjectively confirmed. Prevalence and odds ratios of arterial hypertension and myocardial infarction were computed for subjects who were very much and extremely annoyed by noise, or moderately annoyed, using residents who were slightly annoyed or not annoyed at all as a referent category. Significant odds ratios (adjusted for age, body mass index and smoking habits) were found for self reported arterial hypertension [1,8 (1,0-2,4 - 95% confidence interval), P < 0,01] and myocardial infarction [1,7 (1,0-2,9), P < 0,05] in very much or extremely noise disturbed male subjects, compared to those who were not annoyed at all, or were slightly annoyed by noise. The respective odds ratios for females were lower and not statistically significant 1,1 (0,8-1,7) and 1 (0,4-2,0). PMID- 12537840 TI - Cortical Excitations, Cortisol Excretion and Estimation of Tolerable Nightly Over Flights. AB - Noise induces cortisol excretion even below the awakening threshold. This is based upon the existence of very close subcortical central nervous connections between parts of the auditory system (e. g. amygdala) showing typical plasticity effects and the hypothalmic-pituitary-adrenal (HPA)-axis. Repeated noise events (e.g. over-flights during nigh-time) will lead to accumulation of the cortisol concentration in blood. This happens because its time constant of exponential decrease is about 50 to 10 times larger than that one for adrenaline and noradrenaline. A twofold attempt has been made to calculate the cortisol accumulation using an initial value of noise induced small cortisol increase (rounded value 14 ng/ml) at the nightly threshold of beginning vegetative overreaction around 53 dB(A). A mean time-constant of 64 min has been applied based upon experimental studies. Using in a first step the range of minimal and maximal normal cortisol values as border line and taking into account a relation between peak sound pressure level and cortical excitation given by a power function (exponent 0.32, based on evoked potential studies in man) results in a formula to estimate tolerable events during night-time periods (over-flights in a given time range). Examples of results for 8 hours in the night are for instance values of 11 events with 55 dB(A) indoor peak level or 5 events with 75 dB(A) indoor peak level respectively. Those values of tolerable nightly noise events estimated on the basis of physiological processes and peak levels cannot be recalculated as or compared with equivalent sound levels. PMID- 12537841 TI - Is Electronically Amplified Music too Loud? What do Young People Think? AB - Listening to loud music has been associated, in a number of studies, with hearing loss and tinnitus among young people. However an unanswered question is whether or not these same young people want to have their music so loud. In our study 533 young men and 167 young women, in the age group 16 to 25, who were attending a vocational training centre, responded to a questionnaire and volunteered to have their hearing assessed. The questionnaire sought information on listening habits, on the kinds of events attended, on whether the music at these events was too loud or not, and if the respondents considered their hearing had been impaired. Analysis of this data indicated that 79% of the subjects attend discotheques, 52% pop and rock concerts, and 35% techno parties (e.g. raves). A significant number considered the music at these venues was too loud. Some 42% considered this was the case at discos, 35% thought pop and rock concerts too loud, and 39% held a similar view of techno parties. Conversely, fewer than 3% considered sound levels at these events to be too low. On the basis of the response to the questionnaire we estimate that over half the respondents (56.6%) have a sound exposure (L(eq)) from music of over 87 dB(A). It is not surprising therefore that 71% reported that they had suffered tinnitus following attendance at a music event. The hearing capacity of the sample was measured by audiometry. These measurements detected hearing loss in 11% of the 700 individuals tested. However it was not possible to show that the risk of hearing loss increased with increasing exposure to loud music. We conclude that young people neither demand nor require the excessive sound levels typical of most music events. PMID- 12537842 TI - Urban Noise in the City of Sao Paulo, Brazil: An Important Problem of Public Health. AB - OBJECTIVES: To estimate the sound pressure levels in areas with heavy and with local traffic in the city of Sao Paulo and compare the data obtained with the thresholds established by the local law. METHODS: Twenty-eight points in roads with heavy and local traffic were chosen. The measurements were done Monday to Friday, 8 am to 5 pm, using a 2236 Mediator (Bruel & Kjaer) following the International Standard (ref. no. 1996/1982 1,2,3). Each point was measured a number of times to produce a standard error lower than 1 dB. RESULTS: The equivalent continuous A-weighted sound pressure level (Leq) for the roads with heavy traffic ranged from 70.88 to 80.18 dB(A), mean 75.88 dB(A) (95% CI: 74.49; 77.27) and the maximum peak (MaxP) ranged from 102.47 to 108.37 dB(C), mean 105.63 dB(C) (95% CI: 104.59; 106.68). For the roads with local traffic we observed: Leq from 50.82 to 66.88 dB(A), mean 61.11 dB(A) (95% CI: 57.97; 64.26) and MaxP 83.13 to 97.33 dB(C), mean 92.81 dB(C) (95% CI: 89.80; 95.82). There was a strong evidence, p < 0,001, in favour of the difference between the two types of traffic roads regarding the sound pressure levels. CONCLUSIONS: The Brazilian Standards establishes 50dB(A) and 70dB(A) as the maximum limits for environmental noise in residential and industrial areas, respectively. The results of this study indicate that the problem of urban noise in the city of Sao Paulo needs to be tackled urgently due to its important public health impact. PMID- 12537843 TI - Environmental Noise and Community in Hong Kong. AB - In order to find out the attitude of the community towards environmental noise, community surveys were conducted over the territory of Hong Kong through telephone sampling. Specific surveys were also carried out for areas previously affected by severe aircraft noise. Main observations on the community's response towards noise are that noise pollution was ranked the third among five selected social concerns (after "air pollution" and "security", and higher than "traffic jam" and "cleanliness"); about 60% of the respondents found the territory "noisy"; the most annoying noise source was "traffic noise"; 40% of people found the most annoying noise not tolerable and that most people affected by noise suffered from "distraction". Nonetheless, many did nothing (e.g. did not complain) against the noise and still preferred an open-window life style. PMID- 12537844 TI - Effects of Occupational Exposure to Mercury or Chlorinated Hydrocarbons on the Auditory Pathway. AB - The purpose of this study was to examine the effects of industrial exposure to mercury and chlorinated hydrocarbons (CH) on the auditory pathway. To this effect, auditory brainstem responses (ABR) were recorded from 40 workers exposed to mercury, 37 workers exposed to CH and from a control group of 36 subjects that were never exposed to neurotoxic substances. The interpeak latency (IPL) of waves I-III, III-V and I-V were measured. The mean duration of exposure to mercury and CH was 15.5 (+6.4) and 15.8 (+7.2) years respectively. The air sample monitoring of mercury was 0.008 mg/m(3) (0.32 of the Threshold Limit Value - TLV(R) as published by ACGIH 2000). The mean average air sample monitoring was found to be 98 ppm for TCE, 12.7 ppm for PCE and 14.4 ppm for TCA which is respectively between 0.28 - 0.51 of the TLV(R) of CH. The mean blood mercury (B-Hg) levels were found to be 0.5mgr% (+0.3mgr%), which is 0.13 of the upper range of the permitted biologic exposure index (BEI) published by the ACGIH 2000. The mean urine samples levels of trichloroacetic acid were between 0.11-0.2 of the permitted BEI for the CH workers. The percent of workers exposed to mercury and CH workers with abnormal prolongation of IPL I-III was higher than the control group (42.5% and 33.8% vs. 18.0% respectively p<0.02). These results are consistent with other studies and show that ABR may provide a sensitive tool for detecting subclinical central neurotoxicity caused by CH and mercury PMID- 12537846 TI - Psychological distress in college-aged women: links with unresolved/preoccupied attachment status and the mediating role of negative mood regulation expectancies. AB - In the present study, associations between state of mind regarding attachment, expectancies for negative mood regulation (NMR), and the frequency of psychological problems were specified in a sample of young women (N = 104) involved in a collegiate setting. State of mind regarding attachment was assessed using the Adult Attachment Interview (George, Kaplan, & Main, 1996), while NMR expectancies and psychological health were measured via self-report instruments. The results supported the study hypotheses. As predicted, preoccupied women displayed less confidence in NMR and reported more psychological problems than secure and dismissing women. Similar findings were documented when contrasting unresolved/insecure and unresolved/secure women. NMR expectancies mediated relations between state of mind regarding attachment and psychological health. The study results have implications for attachment research and interventions with young adult women involved in collegiate settings. PMID- 12537847 TI - Attachment and dysthymia: the contributions of preoccupied attachment and agency of self to depression in women. AB - The present research evaluated a conceptual model that links preoccupied attachment to dysthymic disorder in women. From an original community sample of 420 women, 129 women were identified with depressive symptomatology as assessed by the Centre for Epidemiological Studies Depression scale (CES-D). Twenty-four of these 129 women were diagnosed as dysthymic disordered according to the Structured Clinical Interview for DSM-III-R-non-patient edition (SCID-NP). Attachment patterns were assessed using the Adult Attachment Projective (AAP). The results indicate that preoccupied attachment was associated with dysthymia. Discussion concerns the processes that may underlie the association between preoccupied attachment and depression, with attention to the possibly complicating factor of unresolved mourning. PMID- 12537848 TI - The link between childhood trauma and later violent offending: the application of attachment theory in a probation setting. AB - This case-study identifies the links between an insecure-avoidant pattern of attachment organization, unresolved childhood trauma, emotional detachment, substance misuse and violent offending behaviour in adulthood. It demonstrates the way in which attachment theory may be used to explicate offending behaviour and to assess risk in a forensic setting. Further, the model illustrates that the clinical application of attachment theory in brief, time-limited work may enhance the offender's capacity for narrative intelligibility leading to an integration of dissociated thoughts and emotional affect, and to a concomitant cessation of violent behaviour. The process is intersubjective and involves the modification and updating of maladaptive, perceptually distorted cognitive-affective internal working models. This is accomplished by strengthening the offender's capacity for reflective thought, which, in turn, facilitates the organization and integration of error-correcting information. PMID- 12537849 TI - Attachment representations in 6-year-olds: related longitudinally to the quality of attachment in infancy and mothers' attachment representations. AB - A new method to assess attachment representations in children by applying a story completion procedure in doll play (SCPDP) is presented. Transmission and continuity of attachment were tested in 28 German families by using the Strange Situation procedure (SS) with the mother and her infant, the Adult Attachment Interview (AAI) with the mother when the child is 5 years old, and the attachment representations (SCPDP) of the 6-year-olds. Relying on the twofold distinction (secure/insecure), results revealed a significant continuity of attachment from 1 to 6 years of age, and a correspondence between maternal AAI and child's attachment quality in SS, as well as a correspondence between maternal AAI and the 6-year-olds' attachment representations. Using configural frequency analysis, we found continuity in patterns of security and insecurity when looking across the measures of infant and pre-school attachment and maternal adult attachment. Processes underlying the high match of mothers' and children's attachment representations are discussed. PMID- 12537850 TI - Adult attachment stance and spouses' marital perceptions during the transition to parenthood. AB - The relationship between adult attachment and marital perceptions was examined in couples making the transition to parenthood. The Adult Attachment Interview was administered to spouses prenatally, and marital perceptions were assessed prenatally and 3, 12, and 24 months postnatally. The couples were also observed during a marital interaction task. Spouses were assigned to groups based on their attachment security (secure/insecure) and mean level (low/high) of negative emotional escalation in their marital interactions. A series of two-way (attachment security by negative emotional escalation) repeated multivariate analyses of variance, followed by univariate analyses, revealed that insecure husbands and the wives of insecure husbands exhibited different patterns in their marital perceptions than secure husbands and the wives of secure husbands when in high negative escalation marriages. Insecure husbands and the wives of insecure husbands in high negative escalation marriages reported greater declines in positive marital perceptions or less positive marital perceptions overall across the two-year postnatal period than secure husbands and wives of secure husbands in similarly high conflict marriages. Such differences between secure and insecure husbands (and their wives) were not apparent in low negative escalation marriages. Findings are discussed in terms of the protective effect a secure attachment may convey upon spouses during stressful periods in their marriage. PMID- 12537851 TI - Early attachment security, subsequent maternal sensitivity, and later child development: does continuity in development depend upon continuity of caregiving? AB - In order to test the hypothesis (1) that the most competent 3-year-olds would be those with histories of secure attachment (at 15 months) who subsequently experienced (relatively) high-sensitive mothering (at 24 months), (2) that the least competent children would be those with histories of insecure attachment who subsequently experienced (relatively) low-sensitive mothering, and (3) that those with mixed or inconsistent attachment-sensitivity histories would fall in between, data gathered as part of the NICHD Study of Early Child Care were examined. A-priori tests supported the hypothesis in the case of all five developmental outcomes examined (problem behavior, social competence, expressive language, receptive language, school readiness), though group means did not always rank in the predicted direction. Further planned comparisons of children with mixed attachment-sensitivity histories revealed that, in the case of all outcomes, insecurely attached children who subsequently experienced high sensitive mothering significantly outperformed secure children who subsequently experienced low-sensitive mothering. Follow-up analyses highlighted the role of maternal and family stress in accounting for why some infants who were classified as secure at 15 months experienced low-sensitive mothering at 24 months and why some infants classified as insecure subsequently experienced high-sensitive mothering. Results are discussed with regard to the role of early experience in shaping development. PMID- 12537852 TI - Population based ascertainment of twins and their siblings, born in Western Australia 1980 to 1992, through the construction and validation of a maternally linked database of siblings. AB - This paper describes the creation of a unique maternal identifier for use in the investigation of perinatal, postneonatal and child outcomes in relation to maternal characteristics. All Midwives' records of Western Australian (WA) births were routinely linked to registrations of births and deaths for infants born from 1980 to 1992 inclusive, then linked to WA hospital discharge data and to registries of birth defects and cerebral palsy to create a longitudinal health record for each infant. However, since each birth to a woman was recorded as a separate event, there was no way to identify siblings. Probabilistic record linkage, based on information about the mother, was used for this task. Logical inconsistencies within the data were used to test the validity of the linkages between birth records attributed to each mother. Information about the mother from other epidemiological studies and data abstracted from hospital case notes was also used to validate sibships. Linkage of the records of 310,255 births in WA during that period resulted in the formation of 181,133 sibships of one or more children. Pooling the results of all of the validation methods gave an error of 0.9%. Linkage identified 3678 sibships containing multiple births, and 305 sets of maternal twins. Ascertainment of twins and their siblings for an ongoing twin register, the WA Twin Child Health (WATCH) study, was a natural consequence of this process. PMID- 12537853 TI - Recruitment and attrition in twin register studies of childhood behavior: the example of the Australian Twin ADHD Project. AB - There are a growing number of large-scale initiatives for twin registers of children. The Australian Twin ADHD Project (ATAP) is used to illustrate two key limitations which may arise with such studies, namely (1) the importance of including or possibly excluding families in which one or both twins have significant developmental disability, and (2) the selective failure to recruit and/or the selective attrition of families in which parents and children share behavioral difficulties. Initially ATAP excluded 1 in 6 of families whose twins were enrolled in the volunteer-based Australian Twin Registry (ATR), and as more children with significant problems were identified, these families were sequentially excluded. With longitudinal data over ten years, two points about retention were identified: the difficulty of retaining the twins in late adolescence, and the loss of the families whose twins had more ADHD symptoms. We discuss strategies for limiting the loss of families and for ensuring comparability of data across registers with similar interests but different methods of recruitment and exclusion. PMID- 12537854 TI - The Australian Twin Registry. AB - The Australian Twin Registry (ATR), established in the late 1970s, is a volunteer registry of over 30,000 pairs of Australian twins of all zygosity types and ages unselected for their health or medical history. The ATR does not undertake research itself but acts as facilitator, providing an important national and international resource for medical and scientific researchers across a broad range of disciplines. Its core functions are the maintenance of an up-to-date database containing basic contact details and baseline information, and the management of access to the resource in ways that enhance research capacity within Australia while protecting the rights of twins. The ATR has facilitated more than 200 studies using a variety of designs, including classic biometrical twin and twin family studies, co-twin control studies, intervention studies, longitudinal studies, and studies of issues relevant specifically to twins. These have yielded more than 300 peer-reviewed publications to date. Areas of major research include studies of behavior, musculoskeletal conditions, teeth and face patterns, cardiovascular risk factors, substance abuse, and risk factors for melanoma and breast cancer. Extensive longitudinal data are available for around 10,000 pairs. DNA samples have been obtained from more than 6000 twins. Considerable efforts are devoted to maintaining the commitment of registry members and recruitment. The ATR hopes to secure funding to expand its activities, including the systematic collection of DNA samples, so that it can continue to play a major role in the development of twin research and contribute to the annotation of the human genome. PMID- 12537855 TI - The East Flanders Prospective Twin Survey (EFPTS). AB - The East Flanders Prospective Twin Survey (EFPTS) is a registry of multiple births in the province of East-Flanders, Belgium. It has several unique features: it is population-based and prospective, with the possibility of long-term follow up; the twins (and higher order multiple births) are ascertained at birth; basic perinatal data are recorded; chorion type and zygosity are established; and since 1969 placental biopsies have been taken and frozen at -20 degrees C for later determination of genetic markers. The EFPTS is the only large register that includes placental data and allows differentiation of three subtypes of monozygotic (MZ) twins based on the time of the initial zygotic division: the dichorionic-diamnionic pairs (early, before the fourth day after fertilisation), the monochorionic-diamnionic pairs (intermediate, between the fourth and the seventh day post fertilisation), and the monochorionic-monoamnionic pairs (late, after the eight day post fertilisation). This added a new dimension to didymology (the science of twins; delta iota delta upsilon mu omicron zeta = twins): the timing of twinning. Studies can be initiated on primary biases, those originating "in utero". Such studies may throw new light on the controversy over the validity of the classic twin method, the consequences of early embryological events (before and just after implantation of the embryo), the origin of congenital malformations, the sex proportion of multiples, the gene-environment interactions as far as intrauterine environment is concerned, to name but a few. PMID- 12537856 TI - The etiology of personality function: the University of British Columbia Twin Project. AB - The UBC Twin Project is an on-going study of personality and personality disorder that has collected personality data on approximately 1000 adult volunteer twin pairs. The primary purpose of the study is to develop and validate measures of the major forms of personality disorder as a quantitative alternative to the classificatory system first proposed by the DSM-III using the standard methods of personality and behavioural genetics research. To date, the study has explored the univariate and multivariate relationships between the major personality and personality disorder inventories, and their relationships to other psychopathologies including the mood and anxiety disorders and key psychological variables, such as cognitive ability, attachment and attitudes. The results have several implications for personality and personality disorders, most notably a rationalized diagnostic/nosological system to enhanced measurement instruments that index and reflect the influence of specific genetic and environmental influences. PMID- 12537857 TI - Chinese National Twin Registry as a resource for genetic epidemiologic studies of common and complex diseases in China. AB - Twins, due to their unique genetic and environmental relationships, have provided crucial insight in our understanding of genetic contributions to numerous etiologically complex disorders in developed countries. As the leading cause of death and adult disability, cardio- and cerebrovascular diseases are common in China, followed by cancer. Obesity and psychological disorders are increasing. The overall goal of this program is to develop a resource for genetic epidemiologic studies of these and other common and complex diseases in China. Our initial focus is to delineate the genetic and environmental determinants of vascular diseases in general, coronary artery disease (CAD) and stroke in particular. To date, we have over 4500 twin pairs registered and about 700 twin pairs studied for various metabolic traits (e.g., lipids, glucose, insulin, etc.). The long-term plan of this program is to (1) establish a population-based twin registry from several selected regions in China for future studies of specific common complex diseases; (2) conduct detailed phenotyping for clinical and intermediate traits related to cardiovascular diseases; (3) expand studies of twins to twin families by including their parents, siblings, and offspring for genetic linkage and association studies; and (4) follow up twins in the registry longitudinally. The goals of the program are health education and promotion of healthy behavior, early identification of cases to provide timely medical attention, and the evaluation of long-term effects of identified risk factors. We want to develop collaborations with investigators who have expertise in cancer, psychological disorders, and other disease areas. PMID- 12537858 TI - The Danish Twin Registry: 127 birth cohorts of twins. AB - The Danish Twin Registry is the oldest national twin register in the world, initiated in 1954 by ascertainment of twins born from 1870 to 1910. During a number of studies birth cohorts have been added to the register, and by the recent addition of birth cohorts from 1931 to 1952 the Registry now comprizes 127 birth cohorts of twins from 1870 to 1996, with a total of more than 65,000 twin pairs included. In all cohorts the ascertainment has been population-based and independent of the traits studied, although different procedures of ascertainment have been employed. In the oldest cohorts only twin pairs with both twins surviving to age 6 have been included while from 1931 all ascertained twins are included. The completeness of the ascertainment after adjustment for infant mortality is high, with approximately 90% ascertained up to 1968, and complete ascertainment of all liveborn twin pairs since 1968. The Danish Twin Registry is used as a source for large studies on genetic influence on aging and age-related health problems, normal variation in clinical parameters associated with the metabolic syndrome and cardiovascular diseases, and clinical studies of specific diseases. The combination of survey data with data obtained by linkage to national health related registers enables follow-up studies both of the general twin population and of twins from clinical studies. PMID- 12537859 TI - Genetic and environmental factors in complex diseases: the older Finnish Twin Cohort. AB - In studies on the Finnish Twin Cohort, we investigate genetic and environmental determinants of common, complex diseases, and their behavioral risk factors in Finland, a genetically unique and culturally homogenous population. We have formed the following databases: 1) Like-sexed twin pairs (13,888 pairs of known zygosity) form the older Twin Cohort. They have participated since 1975 in mail surveys, in clinical examinations for subsamples, and have been followed-up for morbidity using national medical registers; 2) The older Twin Cohort was expanded in 1996 to include opposite-sex pairs born 1938-1957 (c. 8000 pairs); 3) Two, new longitudinal studies of adolescent twins and their families, form a complementary, ongoing study base described in more detail in an accompanying article. Genetic and environmental effects vary over the life-span, and only longitudinal studies in genetically informative data sets permits the evaluation of such effects. Finally, the inclusion of DNA-based genetic information in a phenotypically rich family data base will offer a unique resource for research in genetic epidemiology and behavioral medicine. PMID- 12537860 TI - Genetic and environmental factors in health-related behaviors: studies on Finnish twins and twin families. AB - Family, twin and adoption studies have provided evidence for familial and genetic influences on individual differences in disease risk and in human behavior. Attempts to identify individual genes accounting for these differences have not been outstandingly successful to date, and at best, known genes account for only a fraction of the familiality of most traits or diseases. More detailed knowledge of the dynamics of gene action and of specific environmental conditions are needed. Twin and twin-family studies with multiple measurements of risk factors and morbidity over time can permit a much more detailed assessment of the developmental dynamics of disease risk and the unfolding of behavioral risk factors. PMID- 12537861 TI - German Observational Study of Adult Twins (GOSAT): a multimodal investigation of personality, temperament and cognitive ability. AB - The German Observational Study of Adult Twins (GOSAT) is the largest population based observational twin study in Germany to date. Embedded in the Bielefeld Longitudinal Study of Adult Twins (BiLSAT), it addresses the etiology of personality, temperament and cognitive ability in a sample of 300 monozygotic (MZ) and dizygotic (DZ) adult twin pairs between 18 and 70 years of age. A major aim of the GOSAT lies in the utilization of different modes of measurement, (i.e., peer reports and observational data), in addition to self-reports which have been used predominantly in past behavioral genetic research on personality and temperament in adults. Participants completed a full day assessment at the University of Bielefeld including videotaped social interactions and presentations, psychometric intelligence tests and computerized elementary cognitive tasks as well as objective measures and unobtrusive behavior counts. The research design of the GOSAT was devised to reduce the potential impact of systematic rater bias on estimates of genetic and environmental influences to a minimum. In combination with extensive self- and peer report data on key personality and personality related dimensions available from the BiLSAT, the GOSAT provides a rich dataset, which currently includes DNA samples from 80% of its participants. PMID- 12537862 TI - Two Italian twin registers for research in human biology and sport science. AB - Two Italian twin registers are described, the Register of Italian Twin Athletes (RITA) and the Twin Register of Rome (TERRY), developed in recent years at the newly established University of Sport and Movement Sciences of Rome-Foro Italico (IUSM). Ascertainment procedures, database, applications and current prospects are outlined, along with their research focus, which mainly consists of epidemiological and clinical research on the determinants of sport performance as well as on the role of genetic factors versus lifestyle, especially physical activity, in health and aging. PMID- 12537863 TI - The Italian Twin Project: from the personal identification number to a national twin registry. AB - The unique opportunity given by the "fiscal code", an alphanumeric identification with demographic information on any single person residing in Italy, introduced in 1976 by the Ministry of Finance, allowed a database of all potential Italian twins to be created. This database contains up to now name, surname, date and place of birth and home address of about 1,300,000 "possible twins". Even though we estimated an excess of 40% of pseudo-twins, this still is the world's largest twin population ever collected. The database of possible twins is currently used in population-based studies on multiple sclerosis, Alzheimer's disease, celiac disease, and type 1 diabetes. A system is currently being developed for linking the database with data from mortality and cancer registries. In 2001, the Italian Government, through the Ministry of Health, financed a broad national research program on twin studies, including the establishment of a national twin registry. Among all the possible twins, a sample of 500,000 individuals are going to be contacted and we expect to enrol around 120,000 real twin pairs in a formal Twin Registry. According to available financial resources, a sub sample of the enrolled population will be asked to donate DNA. A biological bank from twins will be then implemented, guaranteeing information on future etiological questions regarding genetic and modifiable factors for physical impairment and disability, cancers, cardiovascular diseases and other age related chronic illnesses. PMID- 12537864 TI - A gerontological cohort study of aged twins: the Osaka University Aged Twin Registry. AB - We describe subject recruitment and research results from the Osaka University Aged Twin Registry (OUATR). The research focus of OUATR is the genetic and environmental contributions to physical-cognitive-mental aging which we examined in Japanese twins in later adulthood. Within this large-scale registry (12,000 pairs) of oriental twins born between 1900 and 1935, approximately 10% of participants are MZ twins reared apart from early childhood. Two hundred and fifty pairs have had comprehensive medical examinations, including various blood chemical panels, lymphocyte subtests, WAIS (Wechsler Adult Intelligence Scale), and urine analysis. The future foci of this study are primarily on longevity, decline of cognitive functions with aging, bio-physiological functions, lifestyle and behavior genetics, and psycho-spiritual functions. PMID- 12537865 TI - Seoul Twin Family Study: design, sampling, assessments, and future directions. AB - The Seoul Twin Family Study seeks to investigate genetic and environmental influences on the development of cognitive abilities and other psychological traits among children and adolescents in Seoul, South Korea. In November, 2001 with the assistance of the Seoul Metropolitan Office of Education, we began to solicit all private and public schools in Seoul to obtain names, addresses, and telephone numbers of the twins enrolled in schools in Seoul. The vast majority of the recruited twins were between 6 and 18 years old. The recruited twins and their non-twin siblings and parents will undergo comprehensive assessment of cognitive abilities, interests, daily activities, personality, and familial and non-familial environments. We plan to reassess the twins and their families every two years as the twins progress through major changes that characterize childhood, adolescence, and adulthood. In addition, medical records of the twins during the prenatal period will be examined to determine the effects of intrauterine environment on individual differences in cognitive abilities and other psychological traits. We also plan to undertake molecular genetic studies to identify genes involved in specific cognitive abilities. PMID- 12537866 TI - The Korean Twin Registry--methods, current stage, and interim results. AB - The Korean Twin Registry is the first nationwide twin study in Korea. We compiled 154,783 twin pairs from existing nation-wide data sources, mainly from address and national health insurance data. The coverage of this registry was almost complete for the twins born since 1970, but less complete as age increased, so that there were only 990 pairs who were born before 1930. The twins' health examination (N = 54,390 persons) and questionnaire (N = 44,546 persons) results were incorporated into the registry, yielding 12,894 and 9074 concordantly informative pairs. Morbidity and mortality outcomes have been followed up since 1990, for most diseases. For preliminary analysis of complex diseases, we selected ventricular septal defects (VSD) in young twins, stomach and colorectal cancers in adult twins. We identified 353 VSDs, 284 stomach cancers, and 116 colorectal cancers among twins. The prevalence rates of cancers, but not that of VSD, were lower in twins than those in population. The difference in the cancer prevalence was marked for twins born before 1926, implying some degree of selection. Like-sex (LS) twins showed familial recurrence risks (lambdaLS) of 41.2 for VSD and 22.4 for colorectal cancers, and 1.74 for stomach cancers. For opposite-sex (OS) twins, we could estimate lambdaOS of 19.8 for VSD only. These results were compatible with previous studies for VSD and colorectal cancers, but not for stomach cancers. Despite the strength in size, availability of health outcomes, and some lifestyle and basic laboratory data, we need accurate zygosity information to improve the validity of the results. PMID- 12537867 TI - Netherlands Twin Register: a focus on longitudinal research. AB - In 1986 we began The Netherlands Twin Register (NTR) by recruiting young twins and multiples a few weeks or months after birth. Currently we register around 50% of all newborn multiples in The Netherlands. Their parents receive a questionnaire at registration and afterwards when the children are 2, 3, 5, 7, 10 and 12 years of age. Teachers are asked to rate the behavior of the children at ages 7, 10 and 12 years. Adolescent and young-adult twins were recruited through City Councils in the early 1990s. These twins, their parents and siblings participate in longitudinal survey studies that include items about health, fertility, lifestyle, addiction, personality and psychopathology, religion, socioeconomic status, and educational attainment. The total number of twins and multiples registered with the NTR is currently over 60,000. Subgroups of twins and siblings take part in studies of cognitive development, brain function and neuropsychological indices of attention processes, and molecular genetic studies of classical and behavioral cardiovascular risk factors. DNA samples are currently collected in selected twin families for two large linkage studies, which aim to find QTLs for anxious depression and for nicotine addiction. Sisters who are mothers of DZ twins contribute DNA samples for a linkage study of DZ twinning. Large cohorts of phenotyped family members from the general population are very valuable for genetic epidemiological studies and permit selection of informative families for gene finding studies. PMID- 12537868 TI - Norwegian Twin Registers and Norwegian twin studies--an overview. AB - The Norwegian Twin Registers include several sets of population-based sub registers, and covers twin pairs born between 1895 and today. Except for the missing birth years 1960 to 1967, the register is almost complete. Most of the register contains information about both same-sexed and opposite-sexed twin pairs, except for twin pairs born between 1946 and 1960, where only same-sexed twins are registered. In a substantial part of the register, information about zygosity is obtained, mainly by a mailed questionnaire and in some cases supported by DNA testing. These are the birth years 1915 to 1960 and the birth years 1967 to 1979. Zygosity information is further obtained in the different twin studies derived from the twin register. In 1990 the whole register was made available in a computerized form. Several twin studies have been derived from the different parts of the register. In this article, studies from the two earliest parts of the register are reviewed and grouped by recruitment specifics. Finally, future plans for the register and twin studies are discussed. PMID- 12537869 TI - Sri Lankan Twin Registry. AB - Sri Lanka is an island with genetic diversity between the five main population groups. Our twin registry is the first in the developing world. Initially, we established a volunteer cohort of 4600 twin pairs through a competition advertised in the media. In addition, we have volunteer cohorts, birth registration-based cohorts through hospitals, and community-based cohorts. There is also a nationwide population-based younger twin cohort (1992-1997) traced through the Department of Birth and Death Registration. Additionally, we have adapted a Zygosity determination questionnaire and validated it. Establishing ethical guidelines for twin research was a priority because the field of bio ethics is at an early stage of development in Sri Lanka. These guidelines were from a developing world perspective. A sister organization, the Multiple Birth Foundation, was formed to cater to twins and their special needs and to represent their interests, and several branches have been formed. We intend to build capacity by establishing a genetic lab and through crosscultural collaboration. Our vision is to establish a multidisciplinary research foundation. Based on our research findings, we plan to build services to cater to needs of twins by working with professionals, statutory services and government policy makers. PMID- 12537870 TI - The Swedish Twin Registry in the third millennium. AB - Since the Swedish Twin Registry was first established in the late 1950s to study the importance of smoking and alcohol consumption on cancer and cardiovascular diseases, it has been expanded and updated on several occasions. The focus has similarly broadened to most common complex diseases. The content of the database is described, ongoing projects based on the registry are summarized, and we review some of the principal findings on aging, cancer and cardiovascular disease that have come from the registry. Ongoing efforts and future plans for the STR are discussed. Among others, we plan blood collection and genotyping to study the genetic bases of complex diseases, a first contact ever with the cohorts born after 1958, and in-depth studies of selected diseases, such as Parkinson's disease and chronic fatigue syndrome. PMID- 12537871 TI - The Swedish Young Male Twins Register: a resource for studying risk factors for cardiovascular disease and insulin resistance. AB - A new Swedish twin register was created in 1998. All male twins born in Sweden 1973-1979 were identified in the Swedish Medical Birth Register. 3566 twins (1783 pairs) who were alive and residents of Sweden in 1997 were included in the new register. Data on birthweight, birthlength, gestational age, and so forth, were obtained from the Medical Birth Register. Record linkage was effected with the Military Service Conscription Register for the years 1991-1999. Anthropometrical data and information on blood pressure were obtained from this latter source. In 1998 a questionnaire was mailed to all twins in the register. Questions were posed about zygosity, socioeconomic conditions, lifestyle factors (physical activity, smoking habits, etc.), body shape and body size. Complete responses were obtained from 2654 twins (1327 pairs). In 2002 the cohort is being followed up by a more extensive mailed questionnaire. Data will be collected about socioeconomic conditions, lifestyle, eating behaviour (TFEQ), body size and body shape. In the next few years genomic DNA will also be collected from buccal mucosa (using brush techniques). PMID- 12537872 TI - A multiple pregnancy register in the north of England. AB - A regional population-based Multiple Pregnancy Register was established in 1998, with the aim of collecting detailed information on multiple pregnancies to enable research into mortality and morbidity in multiples. Multiple pregnancies are notified to the Register as soon as they are detected, irrespective of whether they resulted in a spontaneous abortion, termination of pregnancy or registered birth. Nine hundred and twenty-six twin pregnancies were recorded during 1998-99, giving a twinning rate of 14.8 per 1000 maternities (rate at birth 13.0 per 1000 maternities). Sixty one per cent of twin pregnancies were detected before 13 weeks of gestation. Chorionicity was determined in 82.6% of 849 twin maternities with at least one stillbirth or livebirth. The fetal loss rate before 24 weeks of gestation was 10.5% (194/1852). The perinatal and infant mortality rates were 40.6 per 1000 births and 32.6 per 1000 livebirths respectively. A prospective Multiple Pregnancy Register not only allows monitoring of trends in multiple birth rates and mortality, but also etiological research and long-term follow-up studies. PMID- 12537873 TI - The St. Thomas' UK Adult Twin Registry. AB - The Registry consists of nearly 10,000 monozygous and dizygous adult caucasian twins aged 18-80 from all over the UK and was started in 1993. This is a volunteer sample recruited by successive media campaigns without selecting for particular diseases or traits. All twins receive a series of disease questionnaires. In addition over half the twins have been assessed in detail clinically for several hundred phenotypes related to common diseases or intermediate traits. The focus has been primarily on cardiovascular, metabolic, musculoskeletal, dermatological, and opthalmological diseases. Over 3000 DZ twins have had a genome wide scan performed as well as many candidate genes allowing both linkage and association studies. The registry has led to many successful innovative research projects, particularly in common diseases previously thought to be predominantly environmental and helped positionally clone some novel genes for common diseases. PMID- 12537874 TI - Twins early development study (TEDS): a multivariate, longitudinal genetic investigation of language, cognition and behavior problems in childhood. AB - The Twins Early Development Study (TEDS) focuses on the early development of the three most common psychological problems in childhood: communication disorders, mild mental impairment and behavior problems. The TEDS twins were assessed longitudinally at 2, 3, 4 and 7 years of age in order to investigate genetic and environmental contributions to change and continuity in language and cognitive development; it is multivariate in order to examine the origins of comorbidity; and it uses a large sample in order to study abnormal development in the context of normal development. The twins were identified from birth records of twins born in the UK in 1994-96. More than 15,000 pairs of twins have been enrolled in TEDS and the participating families are representative of the UK. The measures at 2, 3 and 4 years are administered by parents. At 7 years, children are assessed for language and cognitive development using telephone testing, parents and children are interviewed about behavior problems, and teachers also assess behavior problems as well as academic achievement. One set of findings is that the same genes largely contribute to both language and cognitive problems and the same genes affect normal and abnormal development, a result that suggests that general impairment may be a better target for genetic research than specific language impairment independent of nonverbal cognitive problems. DNA has been obtained so far for more than 4000 pairs and is being used initially in molecular genetic studies of language problems and hyperactivity. PMID- 12537875 TI - The Mid-Atlantic Twin Registry. AB - The Mid-Atlantic Twin Registry (MATR) is a population-based registry of twin pairs ascertained from birth records and school system records of Virginia, North Carolina, and South Carolina. The MATR was formed in 1997 with the merging of the Virginia and North Carolina Twin Registries, and it expanded to include South Carolina when access to twin birth records in that state was granted in 1998. Registered twins ("participants") number more than 51,000, with approximately 46,000 of these individuals representing complete pairs. Roughly two-thirds of MATR participants are over age 18, with a mean age of approximately 35 years. These participants have primarily been drawn from the more than 170,000 identical and fraternal twin pairs born in the three states between 1913 and 2000. Twins and their family members have participated in numerous research projects, ranging from general health surveys to studies on specific health topics such as cardiovascular disease; depression and anxiety; seizures; behavioral development; pregnancy complications; conduct disorder; drug use, abuse, and dependence; cleft lip/palate; obesity; and chronic fatigue syndrome. The MATR has established a privacy policy and strict standard operating procedures to protect the confidentiality of participant data. The MATR considers a limited number of qualified requests per year from investigators interested in recruiting MATR participants into their research studies. PMID- 12537876 TI - The Southern California Twin Register at the University of Southern California. AB - The Southern California Twin Register is the result of an effort to recruit twins of all ages in the city of Los Angeles and surrounding areas. The register currently includes an ethnically diverse sample of more than 2600 twin pairs. The most recently recruited pairs have been drawn primarily from computerized records of enrollments in local public school districts, and are comparable in sex and ethnic distributions to the general public school population. An ongoing twin study of social and moral development in children, based on 600 pairs of 9-year old twins from this register, is also described briefly in this paper. PMID- 12537877 TI - The occurrence of chronic disease and other conditions in a large population based cohort of native Californian twins. AB - We describe the prevalence of chronic diseases and conditions in a large cohort of twins, which has been developed to facilitate studies of the role of genetics and environment in the development of disease. The California Twin Program (CTP) comprises twins born in California between 1908 and 1982. Birth records from all multiple births (256,616 in total) were linked (multiple times between 1990 and 2001) with the California Department of Motor Vehicles (DMV) roster of licensees to obtain address information. The linkages have revealed 161,109 matches and, because of less complete DMV records in some years, were less successful in older females than in all others. To date over 51,000 of these twins have completed a detailed 16-page mailed risk factor questionnaire. Based on estimates of numbers of individuals receiving a questionnaire, our crude response rates are as high as 63.6% (among females currently in their 50s), with an overall crude response rate of 37.9%. Similar to our previous report regarding the first 42,000 twins, the current group who have completed the questionnaire are representative of the population from which they were drawn (in terms of age, sex, race and residential distribution). Self-reported disease frequencies are provided, along with current estimates of future cancer incidence and mortality rates likely to be observed in the group. We outline our plans for cohort expansion, additional studies using the cohort, and future plans for inviting collaboration. PMID- 12537878 TI - Preschool social and cognitive behaviors: the Southern Illinois Twins. AB - The Southern Illinois Twins registry consists of a small group of twins from the southern Illinois area and some areas in Missouri and Kentucky, as well as siblings of twins and singletons who have been involved in the study. The current project has as its primary focus the identification of variables that predict aggressive and prosocial behaviors in preschoolers, especially to explore the etiology of individual differences in aggressive and prosocial behaviors from a behavior genetic perspective. Twins are tested yearly within months of their birthdays, from ages 1 year to 5 years. Siblings are tested at 5 years of age. At ages 1-4 years, children are tested on a cognitive battery and mother-child interactions are videotaped. At 5 years, children engage in a peer play situation in the lab. Results thus far suggest that aggression during play in the lab shows genetic influence, corroborating earlier parent rating studies. PMID- 12537879 TI - Project Grow-2-Gether: a study of the genetic and environmental influences on child eating and obesity. AB - "Project Grow-2-Gether" is a child nutrition study of same-sex, 3- to 7-year-old monozygotic and dizygotic twin pairs. The study attempts to bridge two bodies of literature that have rarely interfaced with respect to obesity and ingestive behavior: the first being behavioral genetic approaches to obesity-related traits, and the second being developmental approaches focusing on parent-child relationships. The overarching aim of Project Grow-2-Gether is to disentangle genetic from potential home-environmental influences on child eating behavior and body fat. This paper reviews the rationale for Project Grow-2-Gether, its procedures, and core phenotypic measurement battery. A focus of the study is acquisition of controlled food intake measurements obtained in the laboratory, measurement of specific home environmental variables, and multi-method evaluation of parent-child feeding relations. Future directions may involve longitudinal assessment of child growth and molecular analyses for specific genes that influence child eating behavior. PMID- 12537880 TI - The Vietnam Era Twin Registry. AB - The Vietnam Era Twin (VET) Registry is composed of 7369 middle-aged male-male twin pairs both of whom served in the military during the time of the Vietnam conflict (1965-1975). The Registry is a United States Department of Veterans Affairs resource that was originally constructed from military records; the Registry has been in existence for more than 15 years. It is one of the largest national twin registries in the US and currently has subjects living in all 50 states. Initially formed to address questions about the long-term health effects of service in Vietnam the Registry has evolved into a resource for genetic epidemiologic studies of mental and physical health conditions. The management and administration of the VET Registry is described with particular attention given to the processes involved with database maintenance and study coordination. Several waves of mail and telephone surveys have collected a wealth of health related information on Registry twins. More recent data collection efforts have focused on specific sets of twin pairs and conducted detailed clinical or laboratory testing. New Registry initiatives for the future include the construction of a web site and the development of a DNA repository. PMID- 12537881 TI - Minnesota Twin Family Study. AB - The Minnesota Twin Family Study is a longitudinal study of 11-year-old and 17 year-old twins and their parents designed to examine factors related to the etiology of substance abuse and related problems. At study intake, the twins and their parents participate in a day-long assessment in our laboratory that includes measures of endophenotypes (e.g., event-related potentials, EEG, autonomic nervous system reactivity, startle eye-blink), psychopathology, personality, cognitive ability, anthropometry, and environmental risk/protective factors. DNA derived from blood is also collected. A parallel longitudinal study of adolescent adoptive siblings, biologically related siblings, and their parents is also underway. Over 1500 twin families and 350 adoptive and biological sibling families have already entered the longitudinal phase of the study. This article provides an overview of study methods, highlights published findings, and describes procedures in place to foster collaboration with other investigators. PMID- 12537882 TI - The Minnesota Twin Registry: current status and future directions. AB - The Minnesota Twin Registry is a birth-record-based twin registry. Begun in 1983, it includes data for 4307 surviving intact pairs born in Minnesota between 1936 and 1955. In addition, the Registry includes 901 twin pairs born in Minnesota from 1904 to 1934, as well as 391 male pairs born in Minnesota from 1961 to 1964. The research focus is primarily on human individual differences assessed by self report. Questionnaires completed by the participants include measures of personality, occupational interests, demographics, and leisure-time activities. We outline major contributions that have resulted from Registry research, as well as current and future research directions. PMID- 12537883 TI - The NAS-NRC Twin Registry of WWII military veteran twins. National Academy of Sciences-National Research Council. AB - The NAS-NRC Twin Registry is one of the oldest, national population based-twin registries in the United States. It consists of 15,924 white male twin pairs born in the years 1917-1927 (inclusive) both of whom served in the armed forces. The registry, which has been in operation more than 30 years, has collected data from a variety of sources. Records-based, computerized data have come largely from the Department of Veterans Affairs, and there have been three major epidemiologic questionnaires, undertaken roughly every 15 years. Classic twin studies on a variety of medical conditions were the early focus of the registry, which now has a strong focus on chronic disease epidemiology. Work on a DNA specimen bank has been proceeding slowly, but is now a top priority, due to the increasing force of mortality in this twin cohort. PMID- 12537884 TI - The Georgia Cardiovascular Twin Study. AB - The Georgia Cardiovascular Twin Study is a longitudinal study of biobehavioral antecedents of cardiovascular disease in youth. It includes roughly equal numbers of African Americans and European Americans, with a total of > 500 twin pairs. Focus of the study is the change in relative influence of genetic and environmental factors on development of risk factors for cardiovascular disease. Future work will explore the influence of polymorphic variation in candidate genes and their potential interaction with the environment on these risk factors. PMID- 12537885 TI - Pittsburgh Registry of Infant Multiplets (PRIM). AB - This paper describes the Pittsburgh Registry of Infant Multiplets (PRIM; Pittsburgh, Pennsylvania), the results of pilot research conducted in this registry, and the plans for future studies. The main focus of the registry is on psychological development and the risk for behavioral disorders. Particularly, characteristics associated with antisociality and the risk for substance use disorders (e.g., aggressivity, hyperactivity/impulsivity), as well as language development and other traits (e.g., dental health) are among the research targets. PMID- 12537887 TI - Foreword. PMID- 12537886 TI - Wisconsin Twin Panel. AB - The Wisconsin Twin Panel was initiated in 1994 to serve a study of the development of childhood mood and behavioral disorders. Families who give birth to twins within the state of Wisconsin are recruited within 6 months of the birth. The panel currently supports three ongoing, longitudinal research projects. Research foci include studying epigenetic contributions to emotional, physical, cognitive, and motoric development of infant and toddler twins; physiological concomitants of childhood temperament; and early risk and resiliency factors related to child psychopathology. All three studies include videotaped observational assessments and biological measures. PMID- 12537888 TI - Website discussion forums: results of an Australian project to promote telecommunication in dermatology. AB - Discussion forums on the Internet allow doctors to discuss issues of interest in a rapid, convenient and educational manner. In March 2002, the department of dermatology at the St George Hospital in Sydney launched a national Website with an online discussion forum for dermatologists. Features of the Website forum include the ability to moderate the discussion boards, a choice of public or private discussions and receipt of automatic email notifications. Over three months, three dermatologists posted a total of 13 messages on three different topics. The low rate of participation by dermatologists may have been due to lack of time or familiarity with the technology. Increased promotion to and education of dermatologists who are most likely to use the discussion forum may improve utilization of this means of communication. PMID- 12537889 TI - Predicting success: stakeholder readiness for home telecare diabetic support. AB - Readiness to adopt a new technology is one factor that contributes to the success of a telehealth programme. Since one goal of telehealth is to improve care, it is appropriate to determine its success through a quality-of-care framework that addresses structure, process and outcome. A qualitative case study of home care in the Calgary Health Region in Alberta set out to understand how clients, nurses, physicians and managers perceived their readiness to use video-visits for home care. Focus groups, home visits, and telephone and face-to-face interviews were used to collect data. Readiness to adopt home telecare was compared between groups, as well as with behaviour predicted in the literature. Differences in perceptions were identified among the four participant groups. Clients and managers identified a higher degree of readiness - clients because of the potential to support independence in their homes and managers because of the potential efficiencies in the system. PMID- 12537890 TI - Cardiac picture archiving and communication systems and telecardiology - technologies awaiting adoption. AB - Diagnostic and therapeutic procedures associated with cardiology are heavily supported by diagnostic imaging technology. The management of such images, including radiographs, echocardiography examinations and cardiac angiography studies, requires a suitable means of handling the data. A number of manufacturers are now offering picture archiving and communication systems (PACS) and telecardiology options. These could greatly improve the efficiency of data management for cardiac examinations, including linkage to radiology and hospital information systems and electronic patient records. A barrier to the implementation of cardiac PACS has been the relatively high capital cost. There have also been technical difficulties in implementing a suitable interface. Historical problems have included 'turf wars' between different specialist groups and a reluctance to shift from well established practice patterns. Early cooperative work between radiologists and cardiologists in the development of coronary arteriography has been replaced by contention between cardiologists, radiologists and vascular surgeons, often driven by economic considerations rather than the needs of the patient. At this stage, cardiac PACS and telecardiology have great potential for improving the coordinated care of cardiac patients in Australia. PMID- 12537891 TI - Lessons learned from the Hospital Without Walls project. AB - The Hospital Without Walls is an ongoing ambitious project in home telecare that incorporates research into physiological monitoring, low-power radio communication, database storage of physiological data and methods of viewing clinically relevant information from large quantities of stored data. The system records vital signs from patients in their homes using a body-mounted, two-way radio system and a base station located in the home, which transmits data records to a central recording facility every day or in response to predefined emergency events. The prototype system has successfully undergone preliminary clinical trials, with a particular clinical emphasis on monitoring activity using three axis accelerometers. Our experience with the trial suggests that there are significant differences in the technical design required for a long-term, home monitoring system and one where monitoring takes place in an environment staffed by health professionals. PMID- 12537892 TI - Tele-otology: planning, design, development and implementation. AB - The view of the tympanic membrane of the ear provides the specialist with important information for the assessment of ear disease and for treatment planning. In rural and remote areas, the incidence of ear disease is high but access to ear specialists is limited. As part of a project to deliver tele otoscopy services to remote Western Australia, we have investigated various aspects of a tele-otoscopy system. We found that the MedRX video-otoscope is relatively safe to use and produces images of very good quality. A confident diagnosis could be made from still images compressed to 20 kByte. Video sequences could be compressed to a ratio of 1:300. Furthermore, a software package has been developed, and we are developing a training course for health-care workers in remote areas. PMID- 12537894 TI - Videoconferencing in facilities providing care for elderly people. AB - Two studies were conducted to test the feasibility of delivering care by videoconferencing to facilities providing care for elderly people. Both used equipment connected by ISDN lines at 384 kbit/s. During the first study, no consultations, care plans or assessments were conducted. During the second study, 120 assessments were conducted in just over two weeks, both face to face and by videoconference. Why was one project so successful and the other not? The reasons related to ownership, planning, participants and location. A comparison of the two projects highlights some of the considerations necessary to ensure the success of any telehealth project. PMID- 12537895 TI - The evolution of a successful telemedicine mental health service. AB - Development of telemedicine mental health services in Alberta evolved via a pilot project, the delivery of routine services to a small group of centres and subsequent expansion to a province-wide programme. Success of the service was linked to support for telehealth by the provincial government and consultation between the Alberta Mental Health Board (AMHB) and local stakeholders. Assessments by the AMHB have shown that telepsychiatry is acceptable and sustainable at a realistic cost. However, there are few measures of clinical effectiveness available and none of cost-effectiveness. A detailed economic evaluation of the telemedicine mental health network would now be a major task. The expansion of telemedicine mental health services has increased the expectations of health-care decision makers. In addition, the complexity of the network has increased and new initiatives, such as the use of telepsychology, have been introduced. Management of this successful telehealth programme continues to be time consuming and challenging. PMID- 12537896 TI - Integrated regional services: are working process changes desirable and achievable? AB - In the hospital district of Helsinki and Uusimaa, 32 municipalities with one or more health centres provide primary care to their residents. Legal and organizational barriers between primary care and hospital care impede the continuity of patient care. Integrating primary and secondary care with the aid of information technology may facilitate a virtual electronic patient record, in which the viewing of images and other patient data is possible regardless of the organization that produced them. For example, in one trial, diabetic patients sent their home blood glucose measurements by modem to their health centre. Preliminary observations suggest that they could increase their glucose testing largely because they were able to transmit the results to the database and receive teleconsultations. Also, a picture archiving and communication system (PACS) has been in operation in two clinics of the Helsinki University Central Hospital for over two years and seven hospitals had become filmless by the end of 2001. A regional PACS is planned to be completed by the year 2004. PMID- 12537897 TI - Videoconferencing training for those working with at-risk young people in rural areas of Western Australia. AB - Rural Links is a videoconference training initiative developed for those who work with at-risk young people in remote and rural regions of Western Australia. The training programme was run twice (in parallel) for two groups of participants: 17 workers from the Great Southern and South West regions of Western Australia and 15 workers from the Wheatbelt, Pilbara and Kimberley regions of Western Australia. The programme consisted of seven 2 h sessions presented over 12 weeks. Objectives of the training programme centred on increasing participants' knowledge and confidence in relation to the training topics. The initiative also aimed to enhance consultation between rural youth networks and a metropolitan based youth mental health service (YouthLink). Analyses indicated that there were improvements in workers knowledge and confidence in relation to training topics following participation in the programme. Comparisons of the improvements made by these rural participants, who accessed training via videoconferencing, and metropolitan participants, who accessed training face to face, revealed few significant differences. Rural participants reported high levels of satisfaction, decreased feelings of professional isolation and an increased likelihood of accessing YouthLink for consultative support as a result of completing the Rural Links training programme. PMID- 12537898 TI - Predicting success: stakeholder readiness for home telecare diabetic support. PMID- 12537899 TI - Using the Australian and New Zealand Telehealth Committee framework to evaluate telehealth: identifying conceptual gaps. AB - Telehealth is strongly supported in policy rhetoric as being economically significant to Australia, but evaluation standards have been insufficiently developed to ensure that this is the case. The use of one such evaluation standard, the Australian and New Zealand Telehealth Committee (ANZTC) framework, for telehealth evaluation in Australia makes good sense. However, that framework emphasizes economic and technical considerations at the expense of social contexts. Furthermore, there must be questions about the utility of a framework which, it appears, has been used to evaluate only a single telehealth project in Australia. The combination of the economic rationalism of health-care policy and the technological determinism of a tool model of information and communication technologies (ICTs) can result in evaluations that fail to match the complexities of the intersection of health-care and ICTs. Using the ANZTC framework while at the same time focusing on explaining, rather than just describing, the links between interventions and outcomes seems a reasonable compromise. This involves understanding complex socio-technical networks and relationships, and requires investigators to engage with the gulf between private opinions, public statements and actual behaviour. PMID- 12537900 TI - Telehealth for older patients: the Hong Kong experience. AB - We studied the feasibility, acceptability and cost-effectiveness of using telemedicine to provide geriatric services to residents of nursing homes. A local 200-bed nursing home supported by the community geriatric assessment team (CGAT) participated in a one-year study, during which videoconferencing was used to replace conventional outreach or clinic-based geriatric care. The feasibility of telemedicine was evaluated by participating specialists in a total of 1001 consultations. Other outcome measures included productivity gains, utilization of hospital emergency and inpatient services, and user satisfaction. Telemedicine was adequate for patient care in 60-99% of cases in seven different disciplines. The CGAT was able serve more patients and see them earlier and more frequently. Telemedicine was cheaper than conventional care, and well accepted by health-care professionals as well as clients. Substantial savings were achieved in the study period through a 9% reduction in visits to the hospital emergency department and 11% fewer hospital bed-days. Telemedicine was a feasible means of care delivery to a nursing home and resulted in enhanced productivity and cost-savings. Linking more such institutions to care providers would further increase cost effectiveness. PMID- 12537901 TI - The cost-effectiveness of telehealth in metropolitan hospitals. AB - We established a telehealth link between two inner-city health services about 10 km apart. The video-link between the sites was based on IP videoconferencing; ISDN connections were maintained as a form of system redundancy and to connect to sites beyond the local area network. The cost of a conventional consultation, in which the patient travelled to an outpatient appointment, was A$170. Because existing equipment and telecommunications could be used, the cost of a telehealth consultation was only A$74. During the study approximately 15 teleconsultations were completed and a number of patient transfers were cancelled because they were deemed clinically unnecessary. The preliminary results suggest that the cost savings from telehealth can be substantial. In addition there were major benefits to continuity of patient care and clinical communication within the organization. PMID- 12537902 TI - Controlled trial of discharge planning by video-link in a UK urban mental health service: responses of staff and service users. AB - We examined the use of videoconferencing in a UK urban mental health service for discharge planning within the framework of the Care Programme Approach (CPA). The study was an AB design. Baseline data were collected over three months, before the introduction of the CPA by videoconferencing. Twenty-seven CPA meetings were held in the baseline phase of the study and 23 during the video-link phase. Service users and professional participants were asked to complete the Guy's Communication Questionnaire (GCQ) at the end of the meeting. A total of 204 GCQs were completed (88% of those issued). Responses were compared between the face-to face and video-link conditions. No significant differences were found in satisfaction measures between the two conditions. These data suggest that the video-link medium is acceptable to service users and professionals alike for discharge planning. PMID- 12537903 TI - Teleneurology in Northern Ireland: a success. AB - In March 1998 two neurologists and a professor of telemedicine met to solve a clinical problem - how to get neurological expertise to patients admitted acutely to hospitals which had no resident neurologist. Four years later, two new consultant neurologists were appointed in Northern Ireland with sessions in telemedicine, possibly the first such appointments anywhere in the world. This feat was achieved by combining science with politics. First, we chose the most appropriate telemedicine equipment and bandwidth for our needs. Second, we altered our neurological consultation to use the technology efficiently. Third, we were able to show scientifically that teleneurology was reproducible, feasible in practice, safe, acceptable and cost-effective. This required money to pay for a research fellow and equipment, which was obtained initially from some departmental research funds and a local hospital. Conventional research funding bodies were, on the whole, unhelpful. It was serendipitous that a review of neurology services in Northern Ireland was set up by the Department of Health. This identified our original problem as one that it wanted solved and we were in the fortunate position of being able to offer telemedicine as a tested solution. One final political push was required to get the money released. The result of this juxtaposition of science and politics should bring benefit to our neurological patients and will perhaps help others trying to establish similar projects elsewhere. PMID- 12537904 TI - Telenursing: clinical nurse consultancy for rural paediatric nurses. AB - Videoconferencing is increasingly being accepted as a medium for health-care. Telenursing is in its infancy in Australia but has enormous potential for nursing care in remote areas. The Child and Adolescent Psychological Telemedicine Outreach Service (CAPTOS) began in 1997 and in its first evaluation recommended more support for paediatric nurses. CAPTOS telenursing began as a new initiative in late 2001. The telenursing project aims to link ward nurses to CAPTOS and local community teams, and to provide both clinical consultancy on nursing and interdisciplinary issues and locally based professional development. Telenursing supports nurses via site visits, videoconferencing sessions, an interactive Website and sabbatical opportunities. Telehealth works with existing services to enhance the nursing care of young people with a complex mixture of psychological and physical health problems. PMID- 12537905 TI - Physical outcome measurements via the Internet: reliability at two Internet speeds. AB - We have developed a software application to enable interactive rehabilitation via the Internet. The reliability of the telemedicine application was examined by comparing it with face-to-face assessment. The physical outcome measures assessed were knee range of motion, quadriceps muscle strength, limb girth and an assessment of gait. One therapist performed both in-person and Internet-based measurements of all outcome measures on 20 normal subjects. There was good agreement between the two techniques (the 95% limits of agreement included zero for all the variables studied). Internet assessments were conducted at two bandwidths: ISDN at 128 kbit/s and the telephone network at 17 kbit/s. Bandwidth had no significant influence on any of the measures. This study suggests that Internet-based physiotherapy interventions delivered to the home are suitable for further development. PMID- 12537906 TI - The clinical achievements of a geriatric telehealth project in its first year. AB - We performed a feasibility study to test the validity of conducting two standard cognitive assessments via videoconferencing. There was a high correlation between the scores from a face-to-face assessment and those from a videoconference. A second trial was conducted with patients living in a rural community, examined both face to face and via videoconferencing. Again, the validity and reliability of the assessment tools were demonstrated for videoconferencing. The acceptability of the technology to patients and clinicians was also shown. As a result of the trials and at the request of rural participants, geriatric telehealth services are now being provided to a rural aged care assessment team (ACAT) on a fee-for-service basis. The success of this project is reflected in its senior clinical and academic 'champions', the establishment of a dedicated telehealth resource and the development of protocols. PMID- 12537907 TI - Telehealth policy: looking for global complementarity. AB - Telehealth is gaining acceptance as a tool for bridging the local and global health-care divides. However, integrating telehealth into existing health infrastructures presents a daunting challenge for governments, policy makers, telehealth advocates and health-care workers. The development of specific inter jurisdictional telehealth policies will significantly improve the ability to meet this challenge. In the policy context, one 'success' is the increasing number of jurisdictions addressing policy issues. However, policy decisions have largely been taken in isolation, within individual health institutions, regions, provinces/states or countries. This represents a failure of the current approach. Telehealth, by its very nature, has the ability to transgress existing geo political boundaries. As a consequence, policy in any single jurisdiction may hamper or even cripple the ability of telehealth to fulfil its potential. Commonality - or at least complementarity - of approach to telehealth policy must be encouraged. To achieve this, it is essential to understand the current or anticipated regulatory constraints that may affect telehealth. We have begun a preliminary study of country-specific policy issues. PMID- 12537908 TI - A comparison of telepaediatric activity at two regional hospitals in Queensland. AB - We conducted a 15-month feasibility study of telepaediatrics. A novel service was offered to two hospitals in Queensland (Mackay and Hervey Bay). We used data from all other hospitals throughout the state as the control group. Although both intervention hospitals were provided with the same service, the telepaediatric activity generated and the effect on admissions and outpatient activity were markedly different. There was a significant decrease in the number of patient admissions to Brisbane from the Mackay region. In addition, there was an increase in the number of Mackay patients treated locally (as outpatients). In contrast, little change was observed in Hervey Bay. We assessed whether the observed differences between the two hospitals were due to various factors which influenced the use of the telepaediatric service. These factors included the method of screening patients before transfer to the tertiary centre and the physical distance between each facility and the tertiary centre. We believe that the screening method used for patient referrals was the most important determinant of the use of the telepaediatric service. PMID- 12537909 TI - Low-cost telemedicine in the developing world. AB - The Swinfen Charitable Trust uses digital cameras and email to provide specialist advice to doctors in developing countries. The first telemedicine link was set up in July 1999. By the end of a year there were three links to hospitals in Bangladesh, Nepal and the Solomon Islands. Initially the consultants, all of whom give their advice free of charge, were from the UK, but now are worldwide. At present there are 12 links in operation, including one on Tristan da Cunha, and two links approved and awaiting equipment. The advice given by the consultants has been found to be helpful to the referring doctors and to benefit their patients. Failures have been due to the use of obsolescent equipment, computer viruses, lack of communication with the referring hospital before setting up a link, and referring doctors not chasing up their own referrals. Problems yet to be solved include the unreliability of the Internet, certain medicolegal issues and assessing the quality of medical consultants. In future there will be the problem of managing a rapidly growing telemedicine network. PMID- 12537910 TI - Problem-based learning: a case study in providing e-health education using the Internet. AB - The current trend among many universities is to increase the number of courses available online. However, there are fundamental problems in transferring traditional education courses to virtual formats. Delivering current curricula in an online format does not assist in overcoming the negative effects on student motivation which are inherent in providing information passively. Using problem based learning (PBL) online is a method by which computers can become a tool to encourage active learning among students. The delivery of curricula via goal based scenarios allows students to learn at different rates and can successfully shift online learning from memorization to discovery. This paper reports on a Web based e-health course that has been delivered via PBL for the past 12 months. Thirty distance-learning students undertook postgraduate courses in e-health delivered via the Internet (asynchronous communication). Data collected via online student surveys indicated that the PBL format was both flexible and interesting. PBL has the potential to increase the quality of the educational experience of students in online environments. PMID- 12537911 TI - International distance education and the transition from ISDN to high-bandwidth Internet connectivity. AB - The Thailand Hawaii Assessment of Interactive Healthcare Initiative (THAI-HI) is an international distance-education project between two teaching hospitals in Honolulu and Bangkok that uses videoconferencing over three ISDN lines. A 'morning report' format is used to discuss clinical cases primarily covering infectious disease and critical-care topics. An audience response system is used at both sites to add interactivity. From July 2001 to May 2002, 816 health-care providers attended 20 clinical conferences. Audiences rated the conferences as highly relevant and as having high training value. Since the ISDN connection is expensive, we plan to convert the telecommunications to a high-bandwidth Internet connection. The Honolulu site will use a 45 Mbit/s commercial connection to the Hawaii Intranetwork Consortium, which links to the Abilene Network on the US mainland. The Bangkok hospital will use a 155 Mbit/s wireless optical connection to UNINET Thailand, which has a 45 Mbit/s circuit to Abilene. PMID- 12537912 TI - The diffusion of innovation: factors influencing the uptake of telehealth. AB - The actual use of telehealth, particularly for clinical consultations, has in many cases been less than anticipated. The literature identifies a number of factors or barriers. These include legal issues, technical difficulties, time and convenience, cost and training/familiarity with the equipment. No single factor has been identified as being consistently present. Rogers' diffusion of innovation theory suggests that organizational structures and cultures will affect health professionals' perceptions of telehealth. The introduction of telehealth services affects existing work practices and workflows. We believe that it is necessary to develop strategies for the introduction of telehealth applications which take into account the idiosyncrasies of the health service sector, and the particular structures and cultures of individual organizations within that sector. PMID- 12537913 TI - Logistical aspects of large telemedicine networks. 1: Site directories. AB - We carried out a survey of the site coordinators in a mature telemedicine network of about 200 sites. The site directory contained information about 221 videoconference facilities. There were 191 site coordinators in all (i.e. some coordinators were responsible for more than one site). Of the 221 sites, we were able to contact 87 on first attempt and 155 by the fourth attempt. Thus there were 66 sites (30%) which were not contactable. We asked each site coordinator to describe any videoconference activity that had taken place over the previous five working days. Of the contacted 155 sites, 78 reported some videoconference activity during the period in question. The total reported videoconference activity was 12,800 min during the one-week monitoring period, that is, an estimated 924 h per month. The most common categories of work were education or training (511 h) and management or administration (225 h), which between them accounted for 80% of all reported videoconference activity. Fifty of the 155 sites (32%) reported that the equipment was not located in an area suitable for patient consultations. In addition, 20 sites (13%) volunteered that their videoconferencing facilities were not in working order at the time of the survey. We did not ask this question in the survey, so that this result represents a lower bound for the true number of inoperable systems. PMID- 12537914 TI - Logistical aspects of large telemedicine networks. 2: Measurement of network activity. AB - We carried out a retrospective review of the videoconference activity records in a university-run hospital telemedicine studio. Usage records describing videoconferencing activity in the telemedicine studio were compared with the billing records provided by the telecommunications company. During a seven-month period there were 211 entries in the studio log: 108 calls made from the studio and 103 calls made from a far-end location. We found that 103 calls from a total of 195 calls reported by the telecommunications company were recorded in the usage log. The remaining 92 calls were not recorded, probably for one of several reasons, including: failed calls - a large number of unrecorded calls (57%) lasted for less than 2 min (median 1.6 min); origin of videoconference calls - calls may have been recorded incorrectly in the usage diary (i.e. as being initiated from the far end, when actually initiated from the studio); and human error. Our study showed that manual recording of videoconference activity may not accurately reflect the actual activity taking place. Those responsible for recording and analysing videoconference activity, particularly in large telemedicine networks, should do so with care. PMID- 12537915 TI - Government relations, government regulations: jumping through the hoops. AB - Over the last decade, telehealth in Australia has been primarily facilitated and driven by government funding. The government now has a major policy initiative in online health. However, in pursuing the broad initiative there is a danger that some of the smaller components can get lost, and this is probably what has happened to telehealth. There appear to be a number of steps required if telehealth in Australia is to keep up the pace of development that occurred in the 1990s, as we move into what is now being called the era of e-health, involving broadband Internet health service delivery. This area is changing extremely rapidly and is increasingly migrating away from the public sector in Australia, where most of the developmental work has occurred, and into the private sector. Many of the issues that require consideration within the domain of e-health in Australia are also relevant to other countries. E-health will significantly change the way that health-care is practised in future, and it is clear that it is the human factors that are more difficult to overcome, rather than the technological ones. PMID- 12537917 TI - Telemedicine and doctor-patient communication: a theoretical framework for evaluation. AB - The literature on doctor-patient communication demonstrates that patient, provider and contextual characteristics influence behaviour within medical encounters, which in turn is an important determinant of health outcomes. This paper introduces a conceptual model which posits that telemedicine affects health outcomes through changes in the way doctors and patients communicate with one another. It also proposes that this process depends on the medium through which the consultation takes place, whether in person, over the telephone, via fax or email, or through two-way interactive video. Since participants in two-way interactive video-consultations rarely meet in person, it is particularly important that the effect of substituting video for face-to-face consultations be delineated. PMID- 12537918 TI - Teleconsultations: who uses them and how? AB - A literature survey was carried out to identify papers describing teleconsulting applications. From 1259 potentially relevant articles identified through Medline, 128 articles were selected for review. The majority of these had been published in the Journal of Telemedicine and Telecare (50 articles, or 39%). We analysed different user groups, equipment and implementation issues, and the type of connections. In 101 studies (79%) the teleconsultations were between doctors, in 11 they were between patient and doctor, in seven between patient and nurse, and in nine between nurse and doctor. Studies of consultations between patients and health-care professionals were thus quite rare. Surgery was the most common specialty in which teleconsultation was described. The teleconsultations were realtime or mainly realtime in 72% of articles. In 39% of studies the primary focus was on videoconferencing. The most common means of connection was by ISDN digital lines (38%). There were very few mentions of how to ensure data protection or to maintain patient confidentiality. We conclude that, for the majority of teleconsultation needs, asynchronous communication is the most flexible and cost-effective approach. Realtime videoconferencing can be justified only in particular circumstances. PMID- 12537919 TI - Interdisciplinary telecommunication and expert teleconsultation in diagnostic pathology: present status and future prospects. AB - Telepathology can be performed in an active or a passive manner. There are two main types: interdisciplinary telepathology and expert teleconsultations. Interdisciplinary telepathology involves a pathologist and another specialist (e.g. a radiologist or endoscopist). It is mainly used in an active manner (i.e. online and in realtime) and has three purposes: to ensure correct tissue sampling, to provide additional diagnostic information to the pathologist, or to facilitate collaborative therapeutic or diagnostic decision making between the clinician and pathologist. Expert teleconsultations, typically between two pathologists, are mainly undertaken in passive mode (i.e. offline and using store and-forward techniques). The Internet often serves as a platform for both types of telepathology application. Experience with interdisciplinary telepathology is rare at present and is usually limited to partners at a local institution. The efficiency and accuracy of expert teleconsultations in telepathology have been demonstrated in numerous studies. In future, the application of expert teleconsultation in pathology will expand, with the Internet as the communications medium. Interdisciplinary telepathology is a promising technique to improve diagnostic quality. PMID- 12537920 TI - A four-year study of telephone support for oncology patients using a non supervised call centre. AB - A non-supervised call centre was used to allow oncology patients to contact their personal doctors 24 hours a day. Incoming patient calls to the centre were automatically routed to the doctor's mobile phone or office telephone. Over four years, an average of 407 calls were made each year, by an average of 274 oncology patients. The average number of emergency hospital visits was 24 per year (0.09 per patient), compared with 42 per year (0.16 per patient) during the year before the telephone service began. The average number of outreach visits was 783 (2.9 per patient) per year during the study, in comparison with 722 (2.8 per patient) before it began. The average number of non-emergency hospital admissions was 41 (0.15 per patient) per year in comparison with 42 (0.17 per patient) before the study. Calls were short (typically 3-5 min) and the majority (88%) were made on the patient's initiative. Outreach patients in receipt of chemotherapy were the principal users (making 88% of all calls). Excluding the initial investment cost, the income received was approximately the same as the running costs of the call centre in two of the three years for which data were available. PMID- 12537921 TI - Validation of a telephone cognitive assessment test battery for use in chronic fatigue syndrome. AB - We compared a computerized version of the Cognitive Drug Research (CDR) cognitive assessment test battery and a completely automated telephone version of the same battery. These assessed aspects of attention, working memory and long-term memory. Both methods were used to assess the cognitive performance of a cohort of 30 people with confirmed chronic fatigue syndrome (CFS) and a group of 30 healthy controls matched for age and education. The CFS group had significantly slower reaction times on all four cognitive measures on both the computerized and telephone tests. The mood data followed similar patterns inthe computer and telephone assessments. The results from both forms of the test battery confirmed the pattern and severity of cognitive impairment in CFS. Furthermore, the two methods of testing were similarly sensitive in detecting cognitive deficits. The incapacitating nature of CFS may cause problems for researchers if the restrictions to mobility affect the representativeness of the study group. The findings of the present study support the use of a fully automated telephone cognitive testing system for detecting deficits in CFS. PMID- 12537922 TI - Telemedicine improves diagnosis of essential hypertension compared with usual care. AB - A randomized controlled trial was conducted to assess the efficacy of a telemedicine service for the diagnosis of essential hypertension. The telemedicine service consisted of using an automatic home blood pressure monitor connected to an ordinary telephone line for the transmission of the data to a central computer. After use of the home monitor for a week, the results were converted to a report form and faxed to the patient's physician. The gold standard for assessment of true hypertension status was 24 h ambulatory blood pressure monitoring. A total of 74 patients from outpatient clinics were randomized into either the telemedicine service or usual care. Use of the telemedicine service significantly improved the detection of essential hypertension compared with usual care: in the telemedicine group, 64% of patients with essential hypertension were diagnosed; in the usual care group, 26% of patients with essential hypertension were diagnosed. Furthermore, diagnosis occurred earlier in the telemedicine group than in patients receiving usual care. Specificity and positive predictive value were similar in the two groups. PMID- 12537923 TI - Transfer of knowledge from the specialist to the generalist by videoconferencing: effect on diabetes care. AB - We conducted a 12-month prospective interventional study of videoconferencing between primary and secondary care. A treatment network consisting of a diabetes specialist and four general practitioners was established. The communications medium was PC-based videoconferencing via ISDN at 128 kbit/s. A total of 154 type 2 diabetic patients entered the study. The specialist was contacted 94 times via videoconferencing. Metabolic and haemodynamic parameters were significantly improved over the course of the study: the mean HbA1c level fell from 8.1% to 7.8%, systolic blood pressure from 156 to 148 mmHg and diastolic blood pressure from 88 to 83 mmHg. The study demonstrated that therapeutic counselling by videoconferencing is feasible in diabetes care and suggests that it reduces hospital admissions and improves the quality of care. PMID- 12537924 TI - Teleneuroradiology: a case from Nepal with clinical and educational benefit. PMID- 12537925 TI - Feasibility of personal prothrombin time measurement in anticoagulant treatment follow-up. PMID- 12537927 TI - Corrigendum. PMID- 12537926 TI - Telediagnosis of 168 cases of renal disease. PMID- 12537928 TI - 2002 summary statistics and acknowledgements. PMID- 12537929 TI - Final report on the safety assessment of Acrylates Copolymer and 33 related cosmetic ingredients. PMID- 12537931 TI - Final report on the safety assessment of Tocopherol, Tocopheryl Acetate, Tocopheryl Linoleate, Tocopheryl Linoleate/Oleate, Tocopheryl Nicotinate, Tocopheryl Succinate, Dioleyl Tocopheryl Methylsilanol, Potassium Ascorbyl Tocopheryl Phosphate, and Tocophersolan. PMID- 12537932 TI - Progress in the imaging of the prostate gland. PMID- 12537933 TI - Monoclonal antibody (MAB) in patients with advanced cancer. PMID- 12537934 TI - Laparoscopic partial nephrectomy and minimally invasive nephron-sparing surgery. AB - Surgical extirpation remains the most effective therapy for renal cell carcinoma. The surgical management of renal masses has evolved away from radical nephrectomy and now includes nephron-sparing surgery for small tumors. Nephron-sparing surgery has similar cure rates and does not appear to compromise cancer control. As the detection of small renal masses by widespread abdominal imaging continues to increase, so will the demand for minimally invasive nephron-sparing procedures. Despite progress in surgical techniques, laparoscopic partial nephrectomy remains a technically challenging procedure. In this review, we discuss the challenges and recent advances in laparoscopic partial nephrectomy and other minimally invasive approaches to renal masses. PMID- 12537935 TI - Should there be a size limit for elective nephron-sparing surgery? AB - Nephron-sparing surgery (NSS) is a mandatory procedure for patients with solid renal masses who also have coexisting urologic or medical conditions that pose a threat to overall renal function. The excellent results observed with this procedure have led patients with normal contralateral kidneys to choose elective NSS as a treatment modality. However, the optimal selection criteria for NSS have not yet been defined. We review the developments in and recent results of NSS and discuss features critical in selecting patients for this procedure on an elective basis. Current data clearly support the use of elective NSS for localized solid renal masses under 4 cm in size. PMID- 12537936 TI - Update on oncocytoma. AB - Oncocytoma is the most common benign solid renal tumor, comprising roughly 5% of resected renal masses. Typically discovered incidentally, oncocytoma is generally asymptomatic and very rarely metastasizes; however, multifocal disease and coexistence with renal cell carcinoma can occur. No currently used imaging techniques can reliably distinguish between oncocytoma and malignant lesions; therefore, patients must undergo resection, or in certain circumstances, biopsy, to definitively establish diagnosis. Careful attention to pathologic features and the adjunctive use of immunostains can aid in discriminating oncocytoma from other renal tumors characterized by granular, eosinophilic cytoplasm, especially chromophobe renal cell carcinoma. Nephron-sparing and laparoscopic surgical approaches can be used to treat appropriately selected patients. PMID- 12537937 TI - Cytoreductive nephrectomy in metastatic renal cancer. AB - Recent randomized prospective trials have suggested that cytoreductive nephrectomy confers a survival advantage in patients with metastatic renal cancer who receive immunotherapy with interferon-alfa-2b. It appears that the patients most benefiting from this approach are those with good performance status, lung only metastases, and adequate cardiac and pulmonary reserve. Nonrandomized trials have further suggested that this survival advantage may be extended in patients who receive more aggressive interleukin-2-based immunotherapy. PMID- 12537939 TI - Biomaterials in urology. AB - Biomaterials such as urethral catheters, urethral stents, and ureteral stents are commonly used in patients with urologic disorders. There are currently many different bulk materials and coatings available for the manufacture of urinary tract biomaterials; however, the ideal material has yet to be discovered. Any potential biomaterial must undergo rigorous physical and biocompatibility testing before commercialization and use in humans. Despite significant advances in basic science research involving biocompatibility issues and biofilm formation, infection and encrustation remain associated with the use of biomaterials in the urinary tract, and therefore, limit their long-term use. This review critically evaluates the literature published over the past 12 months, providing an update on the current status of naturally derived and synthetic polymeric biomaterial use in the urinary tract. We focus on urethral catheters, urethral stents, and ureteral stents. We discuss issues of biocompatibility and new approaches to biocompatibility testing, biomaterials currently available for use, new biomaterials and coatings, and novel ureteral stent designs. Finally, we discuss the future of biomaterial use in the urinary tract. PMID- 12537938 TI - Novel concepts in the staging of renal cell carcinoma. AB - The incidence of renal cell carcinoma (RCC) continues to rise steadily; unfortunately, our ability to cure patients with metastatic RCC remains limited. When developing and evaluating new treatment protocols, it is important to consider the role of prognostic factors, often defined as pretreatment features, that are predictive of outcome. The complexity and variability of patients' individual clinical outcome and the recently recognized limitation of conventional staging systems have lead to the formulation of integrated prognostic staging systems. In this review, we discuss the evolution of various clinically relevant integrated staging systems for RCC. PMID- 12537940 TI - Laser tissue soldering: applications in the genitourinary system. AB - Within the past 25 years, lasers have transitioned from merely destructive or ablative tools to those with reconstructive uses. It has been shown that the application of laser thermal energy to tissue will result in welding of the approximated areas. Furthermore, the addition of protein solder and chromophores (tissue soldering) has increased wound tensile strength while decreasing peripheral tissue damage. Laboratory studies have demonstrated application of laser tissue soldering to virtually all components of the genitourinary system. Increasing human experience in recent years has reinforced the success of this technique. PMID- 12537941 TI - Antisense oligonucleotide therapy for urologic tumors. AB - Modulation of gene expression using antisense oligonucleotides has advanced from the laboratory to the clinic. Numerous companies can, at least partially, attribute their success to the development of antisense techniques, and one antisense drug is currently on the market. Antisense compounds have been used in clinical trials that included patients with urologic tumors, mostly directed at proliferation- or apoptosis-related targets. Furthermore, therapeutic inhibition of many new identified genes is being investigated in preclinical tests. This review provides a contemporary overview of current preclinical and clinical antisense oligonucleotide concepts for the treatment of urologic tumors. PMID- 12537946 TI - Transabdominal laparoscopic adrenalectomy. AB - Laparoscopic adrenalectomy has become the standard technique for the surgical removal of the adrenal gland at many centers worldwide. Functional adrenal tumors such as aldosteronoma, glucocorticoid, androgen/estrogen-producing adenomas, and small-to-moderate sized solitary pheochromocytomas are amenable to removal via a laparoscopic approach. The advantages of laparoscopic adrenalectomy over open adrenalectomy are well documented and include a shorter hospital stay, a decrease in postoperative pain, shorter interval between surgery and return to preoperative activity level, and improved cosmesis. Various laparoscopic approaches to the adrenal gland have been described. Among these are the lateral transabdominal, anterior transabdominal, lateral retroperitoneal, and posterior retroperitoneal approaches. Each of these methods has specific advantages and disadvantages. This article reviews the transperitoneal approach to laparoscopic adrenalectomy, and discusses indications, operative technique, and a survey of the available literature. PMID- 12537942 TI - Heat shock proteins in the genitourinary system. AB - Heat shock proteins are ubiquitous molecules that are expressed in all living organisms in response to stress. They have two specific roles: protection against cell damage and modulation of the immune response. Recent studies have dramatically increased our knowledge and understanding of the role of heat shock proteins in the genitourinary system. This review uses a Medline approach to examine the current literature. PMID- 12537948 TI - Embodied experience and linguistic meaning. AB - What role does people's embodied experiences have in their use and understanding of meaning? Most theories in cognitive science view meaning in terms of propositional structures that may be combined to form higher-order complexes in representing the meanings of conversations and texts. A newer approach seeks to capture meaning in terms of high-dimensional semantic space. Both views reflect the idea that meaning is best understood as abstract and disembodied symbols. My aim in this article is to make the case for an embodied view of linguistic meaning. This view provides a challenge to traditional approaches to linguistic meaning (although may not necessarily be entirely incompatible with them). I discuss several new lines of research from both linguistics and psychology that explore the importance of embodied perception and action in people's understanding of words, phrases, and texts. These data provide strong evidence in favor of the idea that significant aspects of thought and language arises from, and is grounded in, embodiment. PMID- 12537947 TI - Adrenal-preserving minimally invasive surgery: the role of laparoscopic partial adrenalectomy, cryosurgery, and radiofrequency ablation of the adrenal gland. AB - Adrenalectomy has become the standard of care for the management of hormonally active adrenal masses. Various surgical therapies have been proposed to excise completely or destroy these adrenal lesions, which may be benign or malignant. New minimally invasive, adrenal-sparing procedures have recently been introduced, among them laparoscopic partial adrenalectomy, cryosurgery, and radiofrequency ablation. These procedures focus on reducing patient morbidity and hastening postoperative recovery while preserving normal adrenal tissue. However, questions remain about the risks and benefits associated with routine application of minimally invasive therapies for adrenal-sparing surgery in terms of complete tumor extirpation. Clearly, more experience and longer follow-up is necessary to validate these procedures. Herein we describe the surgical techniques and early results of treatment with adrenal-sparing surgery. PMID- 12537949 TI - Hot dogs and zavy cats: preschoolers' and adults' expectations about familiar and novel adjectives. AB - In recent years, a growing body of research has begun to examine the processes that underlie young children's acquisition of adjectival meanings. In the present studies, we examined whether preschoolers' willingness to extend adjectives was influenced by the type of property labeled by familiar adjectives (Experiment 1) and by semantic information conveyed in the sentence used to introduce novel adjectives (Experiment 2). In Experiment 1, we examined preschoolers' and adults' expectations about the generalizability of familiar adjectives of three different types: emotional state terms, physiological state terms, and stable trait terms. On each trial, we labeled a target animal with one of the three different types of adjectives and asked whether these terms could apply to a subordinate-level match, a basic-level match, a superordinate-level match, or an inanimate object. Results indicated that 4-year-olds and adults extended the trait terms, but not the emotional or physiological terms, to members of the same basic-level category. In Experiment 2, we presented 4-year-olds and adults with novel adjectives in one of two verb frames: stable ("This X is very daxy") or transient ("This X feels very daxy"). Participants were more likely to extend the novel adjective to subordinate matches if they were in the Stable frame group than if they were in the Transient frame group. These findings are discussed in terms of implications for young children's expectations about familiar and novel adjectives. PMID- 12537950 TI - What compound nouns mean to preschool children. AB - Compound nouns have multiple meanings in English. The purpose of this study was to explore when children know that compound nouns refer to two objects, one ideally interacting with the other (e.g., "fish shoes" are shoes with fish on them, not next to them). Thirty-five English-speaking three- and four-year-old children participated in this study. They were given both a production and comprehension task with novel compound nouns. The results showed that the three year-olds and the four-year-olds were equally likely to produce compounds to name two interacting objects. However, the three-year-olds were less likely than the four-year-olds to understand that a compound referred necessarily to two objects. These results demonstrate that children's knowledge of the meaning of compound nouns is still developing in the preschool years. Three possible interpretations of the mismatch in comprehension and production are discussed: (1). there are modality differences in processing by children, (2). the meaning tapped by production and comprehension is not identical, and (3). the tasks differed in complexity. PMID- 12537951 TI - Compound fracture: the role of semantic transparency and morphological headedness. AB - This paper explores the role of semantic transparency in the representation and processing of English compounds. We focus on the question of whether semantic transparency is best viewed as a property of the entire multimorphemic string or as a property of constituent morphemes. Accordingly, we investigated the processing of English compound nouns that were categorized in terms of the semantic transparency of each of their constituents. Fully transparent such as bedroom are those in which the meanings of each of the constituents are transparently represented in the meaning of the compound as a whole. These compounds were contrasted with compounds such as strawberry, in which only the second constituent is semantically transparent, jailbird, in which only the first constituent is transparent, and hogwash, in which neither constituent is semantically transparent. We propose that significant insights can be achieved through such analysis of the transparency of individual morphemes. The two experiments that we report present evidence that both semantically transparent compounds and semantically opaque compounds show morphological constituency. The semantic transparency of the morphological head (the second constituent in a morphologically right-headed language such as English) was found to play a significant role in overall lexical decision latencies, in patterns of decomposition, and in the effects of stimulus repetition within the experiment. PMID- 12537952 TI - Semantics and semantic errors: implicit access to semantic information from words and nonwords in deep dyslexia. AB - In this paper we describe dissociations of implicit versus explicit access to semantic information in a patient with deep dyslexia. This acquired reading disorder is characterized by the production of morphological (e.g., SLEEP read as SLEEPING) and semantic errors (e.g., HEART read as BLOOD) and consequently provides a potential window into the operation of both aspects of the language system. The deep dyslexic patient in this study (JO) demonstrated implicit semantic access to items in a number of tasks despite the fact that she was unable to correctly read these items aloud. The findings from this study are consistent with a model of lexical deficits that distinguishes between explicit and implicit access to lexical representations on the basis of inhibitory processes. PMID- 12537953 TI - Both concreteness and age-of-acquisition affect reading accuracy but only concreteness affects comprehension in a deep dyslexic patient. AB - As concreteness correlates very highly with the age-of-acquisition (AoA) of words, we attempted to disentangle the effects of these two variables in the oral reading and comprehension performance of the deep dyslexic patient LW. The results of a multiple regression analysis of LW's reading of 217 words showed that both AoA and concreteness affect reading accuracy, with the AoA effect being most apparent for her reading of concrete words. However, concreteness and not AoA affected LW's performance in matching spoken definitions to printed words, both when the distractors were semantically unrelated and when they were related. These data are interpreted in terms of a model of reading in deep dyslexia in which concreteness affects the ease with which semantics are accessed and can activate lexical representations, and AoA affects the ease with which lexical phonology becomes available for spoken word production. PMID- 12537954 TI - Hemispheric asymmetry in the processing of emotional content in word meanings: the effect of current and past depression. AB - We examined hemispheric lateralization of emotion processing by comparing the performance of clinically depressed, previously depressed, and control individuals on a divided visual field task. Participants were asked to make affective valence judgments for each in a series of laterally presented person descriptive adjectives. Study results suggest that the right cerebral hemisphere (RH) is preferentially sensitive to the affective context of language. Among targets presented to the RH, depressed and previously depressed participants were significantly faster and more accurate in their judgments of negative target words, while controls responded more quickly and accurately to positive target words. No such effects were observed for targets presented to the left hemisphere. It is hypothesized that affective sensitivity may result in differences in semantic network organization across individuals who vary in affective experience. PMID- 12537955 TI - Functional imaging of the semantic system: retrieval of sensory-experienced and verbally learned knowledge. AB - This paper considers how functional neuro-imaging can be used to investigate the organization of the semantic system and the limitations associated with this technique. The majority of the functional imaging studies of the semantic system have looked for divisions by varying stimulus category. These studies have led to divergent results and no clear anatomical hypotheses have emerged to account for the dissociations seen in behavioral studies. Only a few functional imaging studies have used task as a variable to differentiate the neural correlates of semantic features more directly. We extend these findings by presenting a new study that contrasts tasks that differentially weight sensory (color and taste) and verbally learned (origin) semantic features. Irrespective of the type of semantic feature retrieved, a common semantic system was activated as demonstrated in many previous studies. In addition, the retrieval of verbally learned, but not sensory-experienced, features enhanced activation in medial and lateral posterior parietal areas. We attribute these "verbally learned" effects to differences in retrieval strategy and conclude that evidence for segregation of semantic features at an anatomical level remains weak. We believe that functional imaging has the potential to increase our understanding of the neuronal infrastructure that sustains semantic processing but progress may require multiple experiments until a consistent explanatory framework emerges. PMID- 12537956 TI - The semantic category effect increases with worsening anomia in Alzheimer's type dementia. AB - A large number of patients (n=72) with probable Alzheimer's type dementia (DAT) and mild cognitive impairment (MCI) carried out a picture naming task which comprised stimuli from biological and nonbiological categories. The results were stratified into five ranges of overall naming ability. Every group except those with scores within the range of elderly normal individuals demonstrated better nonbiological naming than biological naming, an effect which increased with worsening impairment. In general, patients diagnosed with other dementia (n=15) did not fit well within the pattern of the DAT/MCI participants, except those known to have a significant semantic impairment. A category effect favoring nonbiological items appears to be robust and produce a predictable pattern across progressive levels of impairment in AD. PMID- 12537957 TI - Cryopreservation of precision-cut tissue slices for application in drug metabolism research. AB - Cryopreservation of tissue slices greatly facilitates their use in drug metabolism research, leading to efficient use of human organ material and a decrease of laboratory animal use. In the present review, various mechanisms of cryopreservation such as equilibrium slow freezing, rapid freezing and vitrification, and their application to cryopreservation of tissue slices are discussed as well as the viability parameters often used to evaluate the success of cryopreservation. Equilibrium freezing prevents intracellular ice formation by inducing cellular dehydration, but (large) ice crystals are still formed in the interstitial space of the slices. Upon rapid freezing, (small) intra- and extracellular ice crystals are formed which slices from some tissues can resist. Vitrification prevents the formation of both intra- and extracellular ice crystals while an amorphous glass is formed of the slice liquid constituents. To vitrify, however, high molarity solutions of cryoprotectants are required that may be toxic to the slices. The use of mixtures of high molarity of cryoprotectants overcomes this problem. We conclude that vitrification is the approach that most likely will lead to the development of universal cryopreservation methods for tissue slices of various organs from various animal species. In the future this may lead to the formation of a tissue slice bank from which slices can be derived at any desirable time point for in vitro experimentation. PMID- 12537958 TI - Production of reactive oxygen species by toxin T-514 of genus Karwinskia in vitro. AB - In the present study we have analyzed the production of reactive oxygen species by toxin T-514 of the genus Karwinskia in vitro (primary liver cell cultures and microsomes), as well as their possible role in its cytotoxicity. The role of catalase and superoxide dismutase (SOD) as defense mechanisms against oxidative stress was also studied. Freshly isolated hepatocytes or microsomes were exposed to T-514 in the presence or absence of catalase and SOD. Cytotoxicity was determined by methylthiazoltetrazolium (MTT) reduction. Oxidative stress was evaluated by the dichlorofluorescein diacetate (DCFDA) fluorescent probe and the reduction of ferricytochrome c. Exposure of hepatocytes to toxin T-514 for 2-, 4 , 6- and 24-h periods resulted in a time- and concentration-dependent increase in the suppression of mitochondrial metabolic activity. T-514 induced the production of reactive oxygen species in both hepatocytes and microsomes. Catalase and superoxide dismutase had a protective effect against the cytotoxicity of T-514 in hepatocytes and also inhibited the production of oxygen reactive species in microsomes. The results indicate that oxidative stress mediated by reactive intermediates may be a mechanism by which T-514 induces its cytotoxic effect. PMID- 12537959 TI - Effect of whey protein isolate on intracellular glutathione and oxidant-induced cell death in human prostate epithelial cells. AB - Cysteine is the rate-limiting amino acid for synthesis of the ubiquitous antioxidant glutathione (GSH). Bovine whey proteins are rich in cystine, the disulfide form of the amino acid cysteine. The objective of this study was to determine whether enzymatically hydrolyzed whey protein isolate (WPI) could increase intracellular GSH concentrations and protect against oxidant-induced cell death in a human prostate epithelial cell line (designated RWPE-1). Treatment of RWPE-1 cells with hydrolyzed WPI (500 microg/ml) significantly increased intracellular GSH by 64%, compared with control cells receiving no hydrolyzed WPI (P<0.05). A similar increase in GSH was observed with N acetylcysteine (500 microM), a cysteine-donating compound known to elevate intracellular GSH. In contrast, treatment with hydrolyzed sodium caseinate (500 microg/ml), a cystine-poor protein source, did not significantly elevate intracellular GSH. Hydrolyzed WPI (500 microg/ml) significantly protected RWPE-1 cells from oxidant-induced cell death, compared with controls receiving no WPI (P<0.05). The results of this study indicate that WPI can increase GSH synthesis and protect against oxidant-induced cell death in human prostate cells. PMID- 12537960 TI - Cytogenetic evaluation of two nitroimidazole derivatives. AB - 5-Nitroimidazoles are a well-established group of antiprotozoal and antibacterial agents. Thanks to their antimicrobial activity these chemotherapeutic agents inhibit the growth of both anaerobic bacteria and certain anaerobic protozoa such as Trichomonas vaginalis, Entamoeba histolytica and Giardia lamblia. The aim of the present study is to achieve a precise characterization of the genotoxic activity of these compounds and to establish the value of cytogenetic assays in order to determine the effect of these drugs, at therapeutic doses, to settle an improved risk assessment. Two nitroimidazole were studied, metronidazole and ornidazole, at four different concentrations (0.1, 1, 10 and 50 microg/ml of peripheral blood lymphocyte culture). Endpoints analyzed included: mitotic index (MI), replication index (RI), sister chromatid exchange (SCE) and chromosomal aberrations (CA). An analysis of variance test (ANOVA) was performed to evaluate the results. A significant decrease (P<0.0001) in MI as well as an increase in SCE (P<0.0001) and CA (0.0001) frequencies for both drugs was observed. No modifications in RI were found. The results suggest a genotoxic and cytotoxic effect of MTZ and ONZ in human peripheral blood cultures in vitro. PMID- 12537961 TI - Phagocytosis of titanium particles and necrosis in TNF-alpha-resistant mouse sarcoma L929 cells. AB - In the oral cavity, titanium is an excellent biocompatible material. However, it is reported that high ratios of intracellular titanium particles can cause cell apoptosis or necrosis by as-yet unknown mechanisms. The purpose of this study was to investigate the response of tumor necrosis factor alpha (TNF-alpha)-resistant L929 fibroblasts to titanium particles. Cells were cultured in Eagle's medium supplemented with fetal bovine serum and L-glutamine. Titanium particle sizes were less than 9 micro. Cytotoxicity was assayed by a cell counting kit, trypan blue dye exclusion test and lactate dehydrogenase (LDH) leakage. The production of reactive oxygen species (ROS) was detected by a confocal laser scanning microscope (CLSM) using dichlorofluorescein diacetate as a fluorescent probe. Morphology was viewed by a CLSM and with an X-ray microanalyser (XMA). When titanium particles were added to cells, the viability decreased to around 50% at a particle concentration of 2.0%. The number of dead cells and LDH activity in the culture media increased significantly between 1 and 2 days. However, formation of active oxygen species did not occur, since no dichlorofluorescein fluorescence was observed. A scanning electron photomicrograph (SEM) revealed a large number of particles covering or adhering to cellular components in lysed cells compared with flattened control cells attached to the substrate. The XMA showed that the titanium accumulation was coincident with the deformed cell shape. The CLSM also confirmed that particles were within the cells. From these results it was concluded that titanium particles ingested in large quantities into the cell induced necrosis by a pathway other than by producing ROS. PMID- 12537962 TI - Induction of apoptosis by 4-acetyl-12,13-epoxyl-9-trichothecene-3,15-diol from Isaria japonica Yasuda through intracellular reactive oxygen species formation and caspase-3 activation in human leukemia HL-60 cells. AB - Recently we have reported that the trichothecene mycotoxin 4-acetyl-12,13-epoxyl 9-trichothecene-3,15-diol (AETD) from the fruiting bodies of Isaria japonica Yasuda is a potent inducer of apoptosis in human promyelocytic HL-60 cells. The present study aims to characterize the molecular events leading to AETD-induced apoptosis in HL-60 cells. The percentage of apoptotic cells (annexin-V-positive cell population) increased dose- and time-dependently after AETD exposure. Apoptosis of HL-60 cells by AETD was associated with the formation of intracellular reactive oxygen species (ROS), the depletion of intracellular glutathione (GSH) and the activation of caspase-3. Pretreating the cells with the antioxidant N-acetyl-L-cystein (NAC) and the caspase-3 inhibitor Z-DEVD-fmk abrogated AETD-induced apoptosis and caspase-3 activation. NAC blocked intracellular ROS formation and GSH depletion, but Z-DEVD-fmk did not. These results indicate that AETD induces apoptosis in HL-60 cells by causing intracellular ROS formation and GSH depletion followed by the downstream event of caspase-3 activation. PMID- 12537963 TI - Time course of cytochromes P450 decline during rat hepatocyte isolation and culture: effect of L-NAME. AB - The present work describes an isozyme-related effect of collagenase perfusion on hepatocyte microsomal cytochrome (CYP)-dependent monooxygenase activities: CYP 1A1/2-, 2B1/2-, 3A1/2- and 2E1-dependent activities in microsomes from rat hepatocytes after isolation were about 60% of that of liver microsomes, and CYP 4A1-dependent activity was equivalent to liver microsomes. In contrast, the microsomal protein content of the various CYP isoforms was not affected by hepatocyte isolation. This is in accordance with the hypothesis of CYP inactivation during the process of hepatocyte isolation by collagenase digestion. L-NAME (1 mM) was found unable to protect from the decline of CYP-dependent monooxygenase activities following hepatocyte isolation. It is possible that the decrease in glutathione peroxidase activity observed in the presence of L-NAME, namely depression of defense against peroxynitrite, could counteract the beneficial effect of L-NAME on nitric oxide synthesis inhibition. The present work also shows that L-NAME could not avoid the progressive, isoform-specific, most probably turnover-related, decline of CYP proteins and related monooxygenase activities in cultured hepatocytes. Dysregulations in the mechanisms of CYP expression in rat hepatocyte cultures, presently unknown but nitric oxide independent, thus appear to occur in cultured rat hepatocytes. PMID- 12537964 TI - Refinement of the colony-forming unit-megakaryocyte (CFU-MK) assay for its application to pharmaco-toxicological testing. AB - In vitro colony-forming unit (CFU) assays can be used in the evaluation of potentially haematotoxic compounds during preclinical testing. The use of undifferentiated haematopoietic cells, able to proliferate and commit towards a specific blood cell lineage, enable selective toxicity to be detected. We optimized the colony-forming unit-megakaryocyte (CFU-MK) assay for toxicological applications. We used a collagen-based colony-forming assay to examine the sensitivity of four cell types: mononuclear human cord blood cells (CBC), mononuclear human bone marrow cells (BMC), cord blood enriched CD34+CD38- cells, and bone marrow enriched CD34+CD38- cells, to the toxic effects of five drugs known to cause thrombocytopenia in humans. The enrichment of CD34+CD38- cells was achieved by using a negative cell separation technique. Our results showed that a comparable toxicity was detected both with CBC, BMC and CD34+CD38- cells enriched from cord blood, whereas CD34+CD38- cells from bone marrow were more resistant to some drugs. The assay showed a high reproducibility of the endpoint measured (IC(50)), independently of the cell type used and donor source. The present study demonstrates that the refined CFU-MK assay is reproducible and can be used for in vitro toxicology studies with CBC as well as BMC. PMID- 12537965 TI - Antigenotoxic, antimutagenic and ROS scavenging activities of a Rhoeo discolor ethanolic crude extract. AB - Rhoeo discolor is a legendary plant used for treatment of superficial mycoses in Mexican traditional medicine. Despite its extended use, it is not known whether it has side-effects. An ethanolic crude extract from Rhoeo discolor was prepared, its mutagenic capacity was investigated by the Ames test, and its genotoxic activity in primary liver cell cultures using the unscheduled DNA synthesis assay. This extract was not mutagenic when tested with Salmonella typhimurium strains TA97, TA98 and TA100, and it did not elicit unscheduled DNA synthesis in hepatocyte cultures. In addition, we explored the antimutagenic and antigenotoxic activities of the extract and its ROS scavenger behaviour. Our results show that Rhoeo extract is antimutagenic for S. typhimurium strain TA102 pretreated with ROS-generating mutagen norfloxacin in the Ames test, and protects liver cell cultures against diethylnitrosamine induction of unscheduled DNA synthesis even at 1.9 ng per dish, which was the lowest dose tested. A free radical scavenging test was used in order to explore the antioxidant capacity of Rhoeo extract, as compared with three commercial well-known antioxidants quercetin, ascorbic acid and tocopherol. Rhoeo extract showed less radical scavenging effect than quercetin, but similar to that of alpha-tocopherol and more than ascorbic acid. It is important to note that this extract was neither mutagenic in S. typhimurium nor genotoxic in liver cell culture, even at concentrations as high as four- and 166-fold of those needed for maximal antimutagenic or chemoprotective activities, respectively. PMID- 12537966 TI - Discriminating two classes of toxicants through expression analysis of HepG2 cells with DNA arrays. AB - Microarray technology provides a rapid and cost-effective method to associate specific cellular responses with unique gene expression patterns. If characteristic expression patterns of a small number of genes could be associated with drug toxicity, this association may be used for toxicity prediction, and thereby to reduce the need for traditional toxicity testing. To test this hypothesis, we have designed an array composed of 92 known human genes of toxicological interest (including seven housekeeping genes) and eight bacterial controls. HepG2 cells were treated with either ethanol or one of two quinone containing anticancer drugs, mitomycin C or doxorubicin. RNA was isolated from treated and untreated cells, differentially labeled with fluorescent dyes, and then hybridized to the array. Our results show that the expression patterns induced by ethanol and the anticancer drugs are different. Both of the anticancer drugs, but not ethanol had a differential effect on the regulation of several genes, including CYP4F2/3, CYP3A3, TNFRSF6 and CHES1, demonstrating that the two drugs might function through a similar mechanism, which differs from that of ethanol. These results suggest that microarray-based expression analysis may offer a rapid and efficient means for assessing drug toxicity. PMID- 12537967 TI - Homology modelling of human CYP2E1 based on the CYP2C5 crystal structure: investigation of enzyme-substrate and enzyme-inhibitor interactions. AB - The construction of a homology model of human cytochrome P450 2E1 (CYP2E1) is reported, based on the CYP2C5 crystallographic template. A relatively high degree of primary sequence homology (identity=59%), as expected for proteins of the same CYP family, ensured a straightforward generation of the 3-dimensional model due to relatively few deletions and insertions of amino acid residues with respect to the CYP2C5 crystal structure. Probing the CYP2E1 model with typical substrates of the enzyme showed a good agreement with experimental information in the form of positions of metabolism for substrates, and with site-directed mutagenesis data on certain residues. Furthermore, quantitative relationships between substrate binding affinity and various structural parameters associated with the substrate molecules facilitated the formulation of a procedure for estimating relative binding energy and, consequently, K(m) or K(D) values towards the CYP2E1 enzyme. This method has been based on a consideration of the active site interactions between substrates and key amino acid residues lining the haem pocket, together with compound lipophilicity data from partition coefficients. PMID- 12537968 TI - Use of cultured cells of kidney origin to assess specific cytotoxic effects of nephrotoxins. AB - During drug discovery, assessment of renal safety for a compound is important for further development of a candidate drug. In this study, we describe an in vitro cell-based assay capable of discerning nephrotoxicity. Three cell types, two of kidney origin and one of liver origin, were used to examine the effects of nephrotoxins. The cell types were the porcine normal kidney tubular epithelial cell line (LLC-PK1), the primary human renal proximal tubular epithelial cells (hRPTEC) and the human liver cell line (HepG2). Cytotoxicity was measured using a luciferin/luciferase assay that measures cellular ATP levels. Four known nephrotoxins, 4-aminophenol, cisplatin, cyclosporin A and paraquat, were tested in this cell-based assay to evaluate cytotoxicity on drug exposure. Kidney derived LLC-PK1 cells and hRPTECs were found to be sensitive to selected nephrotoxins while liver-derived HepG2 cells were insensitive. Human RPTEC cells obtained from three individual donors demonstrated highly reproducible effects on drug exposure. With respect to drug discovery efforts, integration of the cell models described here are valuable for evaluation of nephrotoxic potentials during lead selection and optimization processes. PMID- 12537969 TI - Inhibition of neurite outgrowth in N2a cells by leptophos and carbaryl: effects on neurofilament heavy chain, GAP-43 and HSP-70. AB - The neurodegenerative properties of the organophosphate ester leptophos (LEP) and the carbamate ester carbaryl (CB), both of which can cause neuropathic effects in animals, were investigated in differentiating mouse N2a neuroblastoma cells. At a sublethal concentration of 3 microM, both LEP and CB were able to inhibit the outgrowth of axon-like processes from N2a cells after only 4 h of exposure. Extracts of cells exposed to LEP showed decreased cross-reactivities with monoclonal antibodies that recognise the neurofilament heavy chain (NFH) and the growth-associated protein GAP-43. However, they exhibited increased cross reactivity with a monoclonal antibody that recognises the heat shock protein HSP 70. In contrast, no changes were noted in the levels of antibody binding in blots of extracts of cells exposed to CB. It is concluded that, although both LEP and CB inhibit the formation of axons in vitro, the early biochemical changes underlying the neurodegenerative effects of the two compounds are different. PMID- 12537970 TI - When what you have is not enough: optimizing cord blood transplantation in adults. PMID- 12537971 TI - Is it still possible to ameliorate the diagnosis of refractory anemia? PMID- 12537972 TI - Salvage therapy in refractory acute myeloid leukemia: prediction of outcome based on analysis of prognostic factors. AB - Acute myeloid leukemia (AML) non-responsive to initial chemotherapy is generally of poor prognosis. High-dose cytarabine (HD-AraC) has been proposed as salvage therapy in combination with amsacrine. The aim of the current study was first to assess the toxicity and the efficacy of such a combination therapy, and secondly to determine prognostic factors allowing to predict whom patients could benefit of such a treatment. Out of 91, 45 patients referred to our institution have been treated by HD-AraC (3 g/m(2)/12h from day 1 to 4) combined with amsacrine (90 mg/m(2) per day from day 5 to 7) as a salvage regimen. Forty-five of the 91 patients (49%, 95% confidence interval (CI): 39-60%) achieved complete remission (CR). Thirty-five patients were refractory to the salvage therapy and 11 patients died from toxicity during aplasia. Median disease-free survival (DFS) was 11.5 months (95% CI: 6-16 months). After CR achievement, 26 patients received consolidation therapy according to the protocol in which they were included. Nineteen patients with an HLA-identical sibling donor underwent allogeneic bone marrow transplantation. At time of analysis, 27 of the 45 patients (60%) who achieved CR have relapsed. Median overall survival (OS) was 7.5 months (95% CI: 6 15 months). There was 12 long survivors (13%). In univariate analysis, initial karyotype was the main prognostic factor as well as in terms of CR achievement (P=0.002) than in terms of DFS (P=0.01) or OS (P=0.009). CR achievement was negatively influenced by higher WHO performance status index (P=0.006), higher LDH level (P=0.02), and higher CD34 expression by leukemic cells (P=0.03) at diagnosis, and presence of circulating blasts (P=0.001), platelet count <80 x 10(9)/l (P=0.0001), and polymorphonuclear (PMN) percentage <30% (P=0.01) at time of starting salvage therapy. DFS was negatively influenced by secondary AML (P=0.01), weight loss > or =5% (P=0.03), and higher white blood cell (WBC) count (P=0.03) at time of diagnosis. Age over 60 years (P=0.002), prior history of toxic exposure (P=0.01), higher CD34 expression (P=0.02), weight loss > or =5% (P=0.006), and WHO performance status index > or =2 (P=0.01) at diagnosis, and platelet count <80 x 10(9)/l (P=0.02) at time of salvage therapy were the main prognostic factors associated with shorter OS. In multivariate analysis, karyotype grouping at diagnosis (P=0.006) and blood count before salvage therapy (P=0.001) were of prognostic value for CR achievement. Karyotype remained of prognostic value for DFS and OS (P=0.007 and <0.0001, respectively).We conclude that HD-AraC combined with amsacrine was as a useful salvage regimen in AML non responding to a first intensive course of chemotherapy. Using objective parameters of proven significance (karyotypic grouping and blood count before salvage), we devised a prognostic system of immediate clinical utility for prognostic stratification and risk-adapted therapeutic choices. Patients with favorable risk cytogenetics and those with intermediate risk cytogenetics and favorable blood count (PMN > or =30%, no circulating blasts, and platelet count > or =80 x 10(9)/l) before salvage therapy had a similar outcome than those achieving CR after only one course of chemotherapy. All other patients displayed a poor outcome. This suggests their orientation at an earlier time to alternate therapeutic programs based on investigational drugs. PMID- 12537973 TI - Hematopoietic recovery after unrelated umbilical cord-blood allogeneic transplantation in adults treated with in vivo stem cell factor (R-MetHuSCF) and filgrastim administration. AB - Transplantation with unrelated umbilical cord blood (UCB) is marked by delayed hematologic recovery. This report summarizes two adults with chronic myelogenous leukemia (CML), who received myeloablative conditioning followed by infusion of a non-expanded single UCB graft. These CML patients were enrolled in a clinical trial incorporating concomitant in vivo administration of stem cell factor (R MetHuSCF) and filgrastim from day of UCB infusion until attained hematopoietic recovery. Each patient engrafted fully with donor UCB, with days to absolute neutrophil count (ANC) >500/microl being 13 and 29 days, respectively. Both patients remain in cytogenetic remission at 28 months follow-up. 'In vivo UCB expansion' with administration of concomitant R-MetHuSCF and filgrastim may facilitate prompt hematologic engraftment. PMID- 12537974 TI - A prognostic impact of separation of refractory cytopenia with multilineage dysplasia and 5q- syndrome from refractory anemia in primary myelodysplastic syndrome. AB - A prognostic impact of WHO classification of myelodysplastic syndrome (MDS) was studied in a group of 103 primary MDS patients with refractory anemia (RA) according to French-American-British (FAB) classification. Median survival of 37 patients with RA according to WHO criteria of 85.2 months was significantly different from that in both 37 patients with refractory cytopenia with multilineage dysplasia (RCMD) (47.0 months, P=0.002) and 29 patients with 5q- abnormality diagnosed by routine chromosome banding (36.2 months, P=0.0002). A more detailed karyotype analysis with fluorescent in situ hybridization (FISH) techniques confirmed 5q deletion as a sole cytogenetic abnormality in only 12 out of 29 patients, in 4 patients 5q- was associated with complex abnormalities involving 5q region, 13 patients had 5q deletion combined with further karyotype abberations outside 5q. No difference in median survival and estimated 3 years survival was observed between RA patients, patients with 5q- syndrome according to WHO morphology criteria and patients with 5q- as a single abnormality confirmed by FISH in contrast to patients with either additional 5q abberations or further karyotype changes not involving 5q. The same difference was also observed in time to 25% of patients evolving to acute myeloid leukemia (AML). Our study confirmed usefulness of separation of RCMD from RA. RCMD represents a poor prognostic subgroup of MDS clearly distinct from pure RA mainly due to short survival connected with progressive bone marrow failure and increased risk of leukemic transformation. We also suggest to define 5q- syndrome as primary MDS of FAB type RA with 5q deletion as a sole cytogenetic abnormality confirmed by FISH analysis. This definition enabled us to discriminate 5q- patients with favorable prognosis similar as in RA from those with poor outcome associated with 5q- combined with complex abnormalities involving either 5q or regions outside 5q. PMID- 12537975 TI - The spectrum of acute lymphoblastic leukemia with mature B-cell phenotype. AB - We showed heterogeneous disease spectrum among 15 acute lymphoblastic leukemia (ALL) cases with mature B-cell phenotype diagnosed over the past 7 years at our institution. Besides those with typical L3 morphology and 8q24 (c-myc) translocation (n=6), there were cases showing L1 or L2 morphology without 8q24 translocation (n=6), unusually large L3 blasts in hyperdiploid clone (n=2) and blastoid variant of mantle cell lymphoma (n=1). The expression of CD5 and cyclin D1 may need to be routinely determined on ALL cases with mature B-cell phenotype and non-L3 morphology to facilitate timely diagnosis of blastoid MCL and institution of suitable management. PMID- 12537976 TI - Clonal cytogenetic abnormalities in bone marrow specimens without clear morphologic evidence of dysplasia: a form fruste of myelodysplasia? AB - Cytogenetic abnormalities suggestive of a myeloid disorder are occasionally observed in the bone marrow (BM) cells of patients with morphologically and immunohistochemically unremarkable marrow aspirates and biopsies. Between 1994 and 2000, 55 such patients were seen at our institution (34 men; median age of 66 years). The indications for BM sampling included unexplained cytopenias (31 patients), staging or follow-up of a lymphoproliferative disorder or a plasma cell dyscrasia (18 patients), or another miscellaneous reason (6 patients). Specific cytogenetic abnormalities included a 20q deletion or monosomy 20 (10 patients), a chromosome 7 deletion (8 patients), +8 (5 patients), del(5q) or a 5q translocation (4 patients), and del(13q) (2 patients). Eleven patients had a complex karyotype. As of January 2002, 23 of the 55 patients were dead; median follow-up for living patients is 20 months. Of the 23 dead patients, 1 died of acute myelogenous leukemia (AML) and 6 of complications related to cytopenias. This study provides support for obtaining cytogenetic studies in patients with unexplained cytopenias if a morphologic explanation for the cytopenias is lacking. Continued follow-up of this heterogeneous cohort and further studies of similar patients will more clearly define the disease processes and prognosis for this constellation of laboratory findings. PMID- 12537977 TI - CPT-11-induced cell death in leukemic cells is not affected by the MDR phenotype. AB - CPT-11 is a topoisomerase I (Topo I) inhibitor which was initially described as active in multi-drug resistance (MDR) tumors. The MDR phenomenon is characterized by the overexpression of efflux pumps which are able to extrude a range of drugs non-related chemical or functionally. In this work, we treated leukemic cells with CPT-11 300 microM at 24h and compared its cytotoxicity with the activity of efflux pumps and with cell cycle phase. Our findings show that CPT-11 has a potent anti-tumor activity in leukemic cells regardless MDR phenotype and the cell cycle phase, suggesting new avenues to be explored in leukemia treatment. PMID- 12537978 TI - The in vitro generation of Ph1+ ALL-specific HLA-A24-restricted cytotoxic T lymphocytes using a synthetic 16 mer minor bcr-abl peptide. AB - Sixteen mer peptide, which spans the junctional region of the acute lymphoid leukemia (ALL)-specific minor bcr-abl fusion protein and contains a motif that can bind to human leukocyte antigen (HLA)-A24, was constructed. We tried to generate Philadelphia chromosome 1 (Ph1) positive ALL-specific cytotoxic T lymphocytes (CTLs) from eight normal HLA-A24+ individuals with peptide-pulsed autologous dendritic cells (DCs). CTLs could be generated from the mononuclear cells (MNCs) of a single donor, which could kill peptide-pulsed autologous DCs and two A24+ ALL lines, while an HLA-A24+ CML line was only weakly killed and unpulsed DCs or the control lines Daudi or K562 were not recognized. Those CTLs consisted predominantly of CD8+ T cells whose cytotoxicity could be neutralized by monoclonal antibodies to HLA-class I or HLA-A24, and also produced interferon (IFN)-gamma after being stimulated with peptide-pulsed DCs. PMID- 12537979 TI - N-(4-Hydroxylphenyl)retinamide (fenretinide, 4-HPR), a retinoid compound with antileukemic and proapoptotic activity in acute lymphoblastic leukemia (ALL). AB - BACKGROUND: Retinoids have been shown to regulate vital cellular processes including cell proliferation, differentiation and apoptosis. N-(4-Hydroxyphenyl) all-trans-retinamide (fenretinide, 4-HPR) is a synthetic ATRA derivative with chemopreventive and cytotoxic activity against various cancer cell lines including myeloid leukemia. Although several modes of action have been postulated, its mechanism of action in hematologic malignancies remains unclear. Furthermore, only limited information exists as to its activity in lymphoid malignancies. METHODS AND RESULTS: To test whether 4-HPR has activity in acute lymphoblastic leukemia (ALL), we first analyzed its antiproliferative effect in five ALL (Z-33, Z-138, Z-119, Z-181, and Jurkat) cell lines. We found that 4-HPR inhibited the proliferation of all cell lines in a dose-dependent manner at concentrations ranging from 1 to 10 microM. We further demonstrated by cell cycle analysis that 5 microM of 4-HPR blocked Z-119 cells in S phase thus preventing their progression through the cycle. Next we tested whether 4-HPR activated the caspase pathway and induced apoptotic cell death. We found that 4-HPR induced apoptosis in Z-119 cells through the activation of caspase-3 and subsequent cleavage of its substrate poly(ADP-ribose) polymerase (PARP). We then asked whether 4-HPR could affect fresh ALL progenitor cells. Therefore, we obtained bone marrow and peripheral blood cells from five patients with newly diagnosed ALL and tested the effect of 4-HPR using the ALL blast colony culture assay. To supplement our results, we also performed the ALL blast assay on one ALL cell line (ALL-1). We found that 4-HPR significantly inhibited ALL colony-forming cell proliferation in a dose-dependent manner. CONCLUSIONS: Our data show that 4-HPR is a potent inhibitor of ALL cell proliferation and that it induces in vitro apoptotic cell death in ALL blasts. Further studies are warranted to establish the in vivo effect of 4-HPR particularly in patients with ALL. PMID- 12537980 TI - Fusion oncogenic tyrosine kinases alter DNA damage and repair after genotoxic treatment: role in drug resistance? AB - Fusion tyrosine kinases (FTKs) such as BCR/ABL, TEL/ABL, TEL/JAK2, TEL/PDGF beta R and NPM/ALK arise from reciprocal chromosomal translocations and cause acute and chronic myelogenous leukemias and non-Hodgkin's lymphoma. Murine hematopoietic growth factor dependent BaF3 cells and cells transformed by FTK (BaF3-FTK) were used to investigate the role of FTKs in response to DNA damage. FTK-transformed cells displayed resistance to genotoxic treatment including gamma radiation and cytostatic agents such as idarubicin and MNNG. More FTK-transformed cells survived genotoxic treatment and were able to proliferate in comparison to parental non-transformed cells. Similar or higher levels of DNA damage was detected in gamma-irradiated in BaF3-FTK cells in comparison to BaF3 parental cells. Idarubicin induced different amounts of DNA damage in various BaF3-FTK cells. All BaF3-FTK cells treated with MNNG displayed significantly more DNA damage in comparison to BaF3 cells. Despite the extent of genotoxic effect BaF3 FTK cells were often able to repair damaged DNA more efficiently that the non transformed counterparts. Inhibition of BCR/ABL kinase activity by STI571 (Gleevec, inatinib mesylate) abrogated the resistance to genotoxic treatment and inhibited DNA repair mechanisms. We hypothesize that facilitation of the DNA repair in FTK-positive cells may contribute to their resistance to genotoxic treatment. PMID- 12537981 TI - Capsaicin inhibits growth of adult T-cell leukemia cells. AB - Human T-cell leukemia virus type 1 (HTLV-1)-associated adult T-cell leukemia (ATL) is resistant to conventional chemotherapy. We examined the in vitro effects of capsaicin, the principal ingredients of red pepper, on three ATL cell lines. Capsaicin treatment inhibited the growth of ATL cells both in dose- and time dependent manner. The inhibitory effect was mainly due to the induction of cell cycle arrest and apoptosis. Capsaicin treatment also induced the degradation of Tax and up-regulation of I kappa-B alpha, resulting in the decrease of nuclear factor (NF)-kappa B/p65 DNA binding activity. In addition, the Bcl-2 level was found to be decreased. Based on these findings, capsaicin may be considered for chemoprevention of ATL. PMID- 12537982 TI - Masked MLL gene rearrangement was disclosed in the clinical course and sequential development of chromosome abnormality in a patient with therapy related acute myelogenous leukemia. AB - Therapy related acute myelogenous leukemia in a 55-year-old Japanese woman is described. She had been treated for a diagnosis of non-Hodgkin's lymphoma 2 years before the onset of the secondary leukemia. She was diagnosed as AML (FAB: M2) with monosomy 7, and successfully treated by an intensive combination chemotherapy followed by an autologous peripheral blood stem cell transplantation. The disease relapsed shortly after the treatment, and the karyotype analysis revealed a complex abnormality accompanied with t(9;11)(p22;q23), however, monosomy 7 was absent. Southern blotting analysis was performed, and MLL rearrangement was evident in both the bone marrow samples obtained at that time and the cryopreserved marrow cells obtained at the onset of the disease. The bone marrow sample stored in a Carnoy solution at the onset was further analyzed, and three karyotype panels showing 45,XX, -7, t(9;11)(p22;q23) were found. Like this situation, a masked MLL rearrangement may have existed in some cases with hematopoietic malignancies, and appear to be disclosed in the clinical course. PMID- 12537983 TI - Intussusceptive angiogenesis--the alternative to capillary sprouting. AB - In contrast to sprouting angiogenesis, which is a well established mode of new blood vessel formation, intussusceptive angiogenesis (IA) is a relatively new concept in vascular biology. It was first discovered in the lung as a means of capillary network growth (intussusceptive microvascular growth). The mechanism consists in the repeated insertion of new slender transcapillary tissue pillars, which subsequently increase in size, thus allowing the capillary network to grow in itself (i.e., by intussusception). It could be shown that IA was present in all organs and species investigated so far, so that it appears to be an ubiquitous phenomenon in vertebrates at least. It was not a surprise therefore to find that IA also played a role in tumour vascularisation. Morphological analysis has yet brought evidence for 6 different modes of pillar formation. They all have in common that, at one time, two endothelial leaflets (e.g. of opposite capillary walls) come into close contact, form new junctional complexes, then thin out to finally give way to the invading interstitial tissue, particularly to fibroblasts, myofibroblasts and pericytes. Once such a transcapillary pillar is formed, it can subsequently grow to the size of a normal intercapillary mesh. The addition of collagen fibrils to the pillar core will stabilize the pillar mechanically. Recent observations allowed to extend the IA concept further: The same structural mechanism of intussusceptive pillar formation was shown to contribute also to the formation of vascular trees (arborisation) and to be involved in vascular remodeling. Although numerous growth factors and receptors have already been suggested as being active in IA, very few hard facts are at present available which would allow to get a comprehensive view of IA regulation. PMID- 12537985 TI - Chemistry and biochemistry of magnesium. PMID- 12537986 TI - Cell physiology of magnesium. PMID- 12537987 TI - Magnesium: nutrition and metabolism. AB - Magnesium is an essential mineral that is needed for a broad variety of physiological functions. The usual daily magnesium uptake with a western diet is sufficient to avoid deficiency but seems not to be high enough to establish high normal serum magnesium concentrations that are protective against various diseases. Changes in magnesium homeostasis mainly concern the extracellular space, as the intracellular magnesium concentration is well regulated and conserved. The extracellular magnesium concentration is primarily regulated by the kidney, the mechanisms of this regulation have been elucidated recently. Due to the growing knowledge about the regulation of extra- and intracellular magnesium concentrations and the effects of changed extracellular magnesium levels the use of magnesium in therapy gains more widespread attention. PMID- 12537988 TI - Role of magnesium in insulin action, diabetes and cardio-metabolic syndrome X. AB - Magnesium (Mg) is one of the most abundant ions present in living cells and its plasma concentration is remarkably constant in healthy subjects. Plasma and intracellular Mg concentrations are tightly regulated by several factors. Among them, insulin seems to be one of the most important. In vitro and in vivo studies have demonstrated that insulin may modulate the shift of Mg from extracellular to intracellular space. Intracellular Mg concentration has also been shown to be effective in modulating insulin action (mainly oxidative glucose metabolism), offset calcium-related excitation-contraction coupling, and decrease smooth cell responsiveness to depolarizing stimuli. A poor intracellular Mg concentration, as found in noninsulin-dependent diabetes mellitus (NIDDM) and in hypertensive patients, may result in a defective tyrosine-kinase activity at the insulin receptor level and exaggerated intracellular calcium concentration. Both events are responsible for the impairment in insulin action and a worsening of insulin resistance in noninsulin-dependent diabetic and hypertensive patients. By contrast, in NIDDM patients daily Mg administration, restoring a more appropriate intracellular Mg concentration, contributes to improve insulin-mediated glucose uptake. The benefits deriving- from daily Mg supplementation in NIDDM patients are further supported by epidemiological studies showing that high daily Mg intake are predictive of a lower incidence of NIDDM. In conclusion, a growing body of studies suggest that intracellular Mg may play a key role in modulating insulin-mediated glucose uptake and vascular tone. We further suggest that a reduced intracellular Mg concentration might be the missing link helping to explain the epidemiological association between NIDDM and hypertension. PMID- 12537989 TI - Magnesium and cardiac arrhythmias. PMID- 12537990 TI - Magnesium and coronary heart disease. PMID- 12537991 TI - Magnesium and heart failure. PMID- 12537992 TI - Role of magnesium in the pathogenesis of hypertension. AB - Human essential hypertension is a complex, multifactorial, quantitative trait under polygenic control. Although the exact etiology is unknown, the fundamental hemodynamic abnormality in hypertension is increased peripheral resistance, due primarily to changes in vascular structure and function. These changes include arterial wall thickening, abnormal vascular tone and endothelial dysfunction and are due to alterations in the biology of the cellular and non-cellular components of the arterial wall. Many of these processes are influenced by magnesium. Small changes in magnesium levels may have significant effects on cardiac excitability and on vascular tone, contractility and reactivity. Accordingly magnesium may be important in the physiological regulation of blood pressure whereas perturbations in cellular magnesium homeostasis could play a role in pathophysiological processes underlying blood pressure elevation. For the most part, epidemiological and experimental studies demonstrate an inverse association between magnesium and blood pressure and support a role for magnesium in the pathogenesis of hypertension. However data from clinical studies have been less convincing and the therapeutic value of magnesium in the prevention and management of essential hypertension remains unclear. In view of the still ill-defined role of magnesium in clinical hypertension, magnesium supplementation is advised in those hypertensive patients who are receiving diuretics, who have resistant or secondary hypertension or who have frank magnesium deficiency. A magnesium-rich diet should be encouraged in the prevention of hypertension, particularly in predisposed communities because of the other advantages of such a diet in prevention. The clinical aspect that has demonstrated the greatest therapeutic potential for magnesium in hypertension, is in the treatment of pre-eclampsia and eclampsia. The present review discusses the role of magnesium in the regulation of vascular function and blood pressure and the implications in mechanisms underlying hypertension. Alterations in magnesium regulation in experimental and clinical hypertension and the potential antihypertensive therapeutic actions of magnesium will also be addressed. PMID- 12537993 TI - Low magnesium and atherosclerosis: an evidence-based link. AB - Several data indicate that magnesium deficiency caused by poor diet and/or errors in its metabolism may be a missing link between diverse cardiovascular risk factors and atherosclerosis. Experimentally induced low plasma levels of magnesium accelerate atherogenesis by increasing LDL concentrations and their oxidative modifications, and by promoting inflammation. In vitro studies have shown that low magnesium determines endothelial dysfunction, the initiating event leading to the formation of the plaque. Moreover, oral magnesium therapy has been shown to improve endothelial function in patients with coronary artery disease.Magnesium, which is an inexpensive, natural and rather safe element, could be useful in preventing atherosclerosis and as an adjuvant therapy in patients with clinical manifestations of the disease. PMID- 12537995 TI - Dantrolene analogues revisited: general synthesis and specific functions capable of discriminating two kinds of Ca2+ release from sarcoplasmic reticulum of mouse skeletal muscle. AB - The general synthesis of dantrolene analogues with various substituents on its phenyl ring has been developed via palladium-catalyzed cross-coupling reactions, the Stille or Suzuki reaction, as the key step. The effects of synthesized analogues have been evaluated by two kinds of Ca(2+) release modes from sarcoplasmic reticulum (SR) of mouse skeletal muscle fibers based on: (1) the measurement of twitch contraction caused by the physiological Ca(2+) release (PCR) of intact skeletal muscle and (2) the rate of Ca(2+)-induced Ca(2+) release (CICR) in saponin-treated skinned muscle fibers. Although dantrolene, a lead compound, inhibits both twitch contraction and CICR, some structurally modified analogues exhibit one or the other of these effects. The methoxy congener, GIF 0185, potently inhibits the twitch contraction without affecting the CICR, while GIF-0166 and GIF-0248, the ortho-nitro regioisomer and ortho, ortho-dinitro substituted analogues, respectively, doubly potentiate the CICR exclusively. PMID- 12537996 TI - Endomorphin 2 analogues containing Dmp residue as an aromatic amino acid surrogate with high mu-opioid receptor affinity and selectivity. AB - To investigate the effectiveness of a 2',6'-dimethylphenylalanine (Dmp) residue as an aromatic amino acid surrogate, endomorphin 2 (EM(2): Tyr-Pro-Phe-Phe-NH(2)) analogues were prepared, in which the constitutive aromatic amino acids (Tyr(1), Phe(3), or Phe(4)) were replaced by Dmp or its isomer, D-Dmp. Replacement of Phe(3) by Dmp increased the affinity over 10-fold for both mu- and delta-opioid receptors, without affecting receptor selectivity. In contrast, replacement of Phe(4) considerably reduced the mu-receptor affinity and selectivity. These data indicated that the Dmp-substitution of Phe(3), but not Phe(4), in EM(2) is favorable for improving mu-receptor specificity. Inversion of the chirality of the substituted Dmp residue resulted in marked decrease in the mu-receptor affinity. Replacement of Tyr(1) by Dmp yielded an analogue that exhibited only a limited decrease in mu-receptor affinity and GPI potency, despite the lack of a phenolic hydroxyl group at the N-terminal residue. In contrast, D-Dmp(1)- or Phe(1)-substitution of Tyr(1) resulted in a significant decrease in mu-receptor affinity and GPI potency. These results suggested that the Dmp residue can mimic Tyr(1), which is one of the critical structural elements of opioid peptides. PMID- 12537997 TI - Syntheses and antiproliferative activities of rebeccamycin analogues bearing two 7-azaindole moieties. AB - As a part of structure-activity relationship studies on rebeccamycin analogues, compounds containing two aza-indole moieties were synthesized bearing either a methyl group or a hydrogen atom on the imide nitrogen. The azaindole substructures were expected to enhance the cytotoxicity toward tumor cell lines through stronger hydrogen bonding with the target enzyme(s). The cytotoxicities of compounds 8, 10 and 19 against a panel of tumor cell lines were examined and compared with those of rebeccamycin, dechlorinated rebeccamycin 2 and N methylated analogue A. Their effect on the L1210 cell cycle was also evaluated. Compound 19, having an imide NH function had the strongest cytotoxicity towards L1210 cells and induced the largest accumulation of cells in the G2+M phases of the cell cycle. In contrast to their non-aza analogues, which were cytotoxic for all the cell lines tested, diaza compounds 10 and 19 showed selectivity for some cell lines. PMID- 12537998 TI - Design and synthesis of orally bioavailable inhibitors of inducible nitric oxide synthase. synthesis and biological evaluation of dihydropyridin-2(1H)-imines and 1,5,6,7-tetrahydro-2H-azepin-2-imines. AB - The process of discovery and biological evaluation of alpha,beta-unsaturated cyclic amidines, as selective inhibitors of inducible nitric oxide synthase (iNOS), is reported. Dihydropyridin-2(1H)-imines and 1,5,6,7-tetrahydro-2H-azepin 2-imines were synthesized and biologically evaluated both in vitro and in vivo using a nitric oxide synthase inhibition assay. Compounds 1, 5, 6, 8-12 and 16 exhibited potent inhibition of iNOS. Among these, compounds 6, 7, 10, 11 and 16 showed 5- to 19-fold isoform selectivity. Compounds 1, 6, 10, 11 and 16 also showed potent inhibitory activity in the NOx accumulation assay in mice. Compounds 1 and 6 showed excellent bioavailability (BA) in rats when administered orally. Full details are presented here, including the structure-activity relationship (SAR) studies, the chemistry of these compounds, and the pharmacokinetic data and the computer-aided docking study of 10 with hiNOS. PMID- 12537999 TI - Biologically active phenols from Saussurea medusa. AB - Sixteen phenolic compounds were isolated from the polar fraction of Saussurea medusa and were structurally elucidated by chemical evidences and spectral methods. These compounds include two new lignan glucosides, namely medusasides A (1) and B (2), and fourteen known phenolic compounds (3-16). One major compound, apigenin 7-O-[alpha-L-rhamnopyranosyl-(1-->6)-beta-D-glucopyranoside] (6) showed remarkable activity to attenuate the scopolamine induced memory deficit of mice. Compound 6 and another major one, quercetin (8) also exhibited moderate cell protecting activities against hydrogen peroxide (H(2)O(2)) induced PC12 cell damage. PMID- 12538000 TI - Effects of sesquiterpenes and amino acid-sesquiterpene conjugates from the roots of Saussurea lappa on inducible nitric oxide synthase and heat shock protein in lipopolysaccharide-activated macrophages. AB - The methanolic extract of the roots of Saussurea lappa CLARKE, a Chinese medicinal herb Saussureae Radix, was found to inhibit nitric oxide (NO) production in lipopolysaccharide (LPS)-activated mouse peritoneal macrophages. Among the constituents from the methanolic extract, two sesquiterpene lactones (costunolide and dehydrocostus lactone) and two amino acid-sesquiterpene conjugates (saussureamines A and B) potently inhibited LPS-induced NO production (IC(50)=1.2-2.8 microM). Saussureamines A and B in addition to costunolide and dehydrocostus lactone did not inhibit iNOS enzyme activity, but they inhibited both induction of inducible NO synthase and activation of nuclear factor-kappaB in accordance with induction of heat shock protein 72. PMID- 12538001 TI - gamma-Carbolines: binding at 5-HT5A serotonin receptors. AB - Screening of various agents resulted in the identification of 5-methyl-1,2,3,4 tetrahydro-gamma-carboline (1; K(i)=5,300 nM) as a compound with modest affinity for mouse 5-HT(5A) receptors. Structure-affinity studies were conducted resulting in 5-methyl-2-[3-(4-fluorophenoxy)propyl]-1,2,3,4-tetrahydro-gamma-carboline (17; K(i)=13 nM). Although 17 also binds at 5-HT(2) receptors, it serves as a novel lead for the further development of 5-HT(5A) ligands. PMID- 12538002 TI - First synthesis of racemic saphenamycin and its enantiomers. investigation of biological activity. AB - The natural antibiotic saphenamycin, 6-[1-(2-hydroxy-6-methyl-benzoyloxy)-ethyl] phenazine-1-carboxylic acid, was synthesized from saphenic acid using temporary allyl protection of carboxy and phenoxy functionalities. Resolution of racemic saphenic acid was performed by crystallization of the corresponding (-)-brucine diastereomeric salts and the absolute configuration of (-)-brucinium (-) saphenate was determined by X-ray crystallography to have R-configuration. This also proved to be the configuration of natural saphenic acid. Enantiomers of saphenamycin were obtained from resolved saphenic acid and screened against a range of skin flora and resistant Staphylococcus aureus strains. Biological activities of saphenamycin enantiomers were compared with that of the synthetic racemate as well as earlier reported activities of saphenamycin isolated from natural sources. No significant difference was observed in activity of the enantiomers of saphenamycin, which revealed that the chirality of saphenamycin has no consequences for the antibiotic activity. Saphenamycin proved to be a potent antibiotic against fusidic acid and rifampicin resistant S. aureus strains showing MIC of 0.1-0.2 microg/mL. PMID- 12538003 TI - The design of potent hydrazones and disulfides as cathepsin S inhibitors. AB - The design and synthesis of dipeptidyl disulfides and dipeptidyl benzoylhydrazones as selective inhibitors of the cysteine protease Cathepsin S are described. These inhibitors were expected to form a slowly reversible covalent adduct of the active site cysteine of Cathepsin S. Formation of the initial adduct was confirmed by mass spectral analysis. The nature and mechanism of these adducts was explored. Kinetic analysis of the benzoyl hydrazones indicate that these inhibitors are acting as irreversible inhibitors of Cathepsin S. Additionally, the benzoylhydrazones were shown to be potent inhibitors of Cathepsin S processing of Class II associated invariant peptide both in vitro and in vivo. PMID- 12538004 TI - Anthranilic acid derivatives: a new class of non-peptide CCK1 receptor antagonists. AB - Having successfully obtained new CCK(1) ligands holding appropriate groups on the anthranilic acid dimer used as molecular scaffold we were interested in increasing their micromolar affinity for the CCK(1) receptors by modifying the spatial relationship of the main pharmacophoric groups. Since, we have proposed simplified analogues reducing the anthranilic acid dimer to a monomer. In this stage of our research program we have prepared and tested on CCK receptors a series of N-substituted anthranilic acid derivatives keeping a Phe residue at the C-terminal site. The indole-2-carbonyl group imparts the best CCK(1) receptor binding affinity (compound 1: IC(50)=197.5 nM) while a sharp decrease in binding affinity is observed for the other indole containing derivatives. Moreover, in order to support the different binding behaviour observed for the synthesized compounds, a conformational investigation was carried out. Finally, on the basis of the main pharmacophoric groups of the obtained new lead compound (1) (coded VL 0395) a receptor binding hypothesis has been provided. PMID- 12538005 TI - Design and synthesis of 3-phenyl tetrahydronaphthalenic derivatives as new selective MT2 melatoninergic ligands. AB - Tetrahydronaphthalenic analogues of melatonin have been synthesized and evaluated as melatonin receptor ligands. Introduction of a phenyl substituent in the 3 position of the tetraline ring allows to obtain MT(2) selective ligands. Activity and MT(2) selectivity can be modulated with suitable modifications of the N-acyl substituent. The (+)-(RR)-cis enantiomer of the N-[2-(7-methoxy-3-phenyl-1,2,3,4 tetrahydro-naphthalen-1-yl)ethyl]cyclobutyl carboxamide (14) is one of the most MT(2) selective ligands described until now and behaves as an antagonist. PMID- 12538006 TI - Synthesis and insecticidal activity of novel N-oxydihydropyrroles: 4-hydroxy-3 mesityl-1-methoxymethoxy derivatives with various substituents at the 5-position. AB - This paper reports the synthesis and insecticidal activity of a series of novel 4 hydroxy-3-mesityl-1-methoxymethoxy-1,5-dihydro-2H-pyrrol-2-one derivatives, in which the substituents at the 5-position were varied with a number of alkyl and spirocycloalkyl groups. Investigation of the structure-activity relationships revealed that small alkyl and spirocyclohexyl groups had a favorable effect on the insecticidal activity of these agents against Myzus persicae. PMID- 12538007 TI - Design and synthesis of 4H-3-(2-phenoxy)phenyl-1,2,4-triazole derivatives as benzodiazepine receptor agonists. AB - A series of new 5-substituted analogues of 4H-3-(2-phenoxy)phenyl-1,2,4-triazole and its chlorinated derivatives was designed and prepared. Conformational analysis and superimposition of energy minima conformers of the compounds on estazolam, a known benzodiazepine receptor agonist, revealed that the main proposed benzodiazepine pharmacophores were well matched. Rotarod and pentylenetetrazole-induced lethal convulsion tests showed that the introduction of an amino group in position 5 of 1,2,4-triazole ring especially in chlorinated derivatives had the best effect which was comparable with diazepam. PMID- 12538008 TI - Syntheses of 3-carbomethoxy-4-(aryl)piperidines and in vitro and in vivo pharmacological evaluation: identification of inhibitors of the human dopamine transporter. AB - A series of 3-carbomethoxy-4-(aryl-substituted)piperidines with various aryl groups were synthesized and examined for binding and reuptake inhibition at the human dopamine transporter, the human serotonin transporter, and the human norepinephrine transporter. The binding potency and reuptake inhibition efficacy was compared with that of (-)-cocaine to determine the significance of removing the two-carbon bridge of the cocaine nucleus on the inhibition of transporter binding and reuptake. Of the transporters examined, the substituted piperidines were relatively selective for the human dopamine transporter. In all cases examined, the cis-diastereomer of the 3-carbomethoxy-4-(aryl substituted)piperidine was observed to be a more potent inhibitor of the human dopamine transporter than the trans diastereomer. Based on the K(i) (binding) and IC(50) (reuptake inhibition) values obtained, the most potent inhibitor of the series was cis-3-carbomethoxy-4-(4'-chlorophenyl)piperidine, and this compound suppressed spontaneous- and cocaine-induced stimulation in non-habituated male Swiss-Webster mice. The conclusion is that substantial portions of the cocaine structure can be dissected away to provide compounds with significant binding and reuptake inhibition of the human dopamine transporter. PMID- 12538009 TI - Rational design and synthesis of peptide ligands for an anti-carbohydrate antibody and their immunochemical characterization. AB - Molecular mimics of carbohydrates present an alternative source of compounds to target pathways involving protein-carbohydrate interactions. Certain peptides act as molecular mimics of carbohydrates in binding to anti-carbohydrate antibodies. A series of potential peptide ligands for the anti-carbohydrate antibody SYA/J6, directed against Shigella flexneri Y, was designed by molecular modeling based on a crystal structure of the antibody complex with a carbohydrate-mimetic peptide. These octapeptides were synthesized using solid-phase peptide synthesis, and their recognition by the antibody was investigated. The results shed light on the nature of peptide-carbohydrate mimicry. PMID- 12538010 TI - Synthesis of new arylalkoxy amido derivatives as melatoninergic ligands. AB - Amido derivatives 10-18 of the corresponding oxyamines were synthesised as melatoninergic ligands by the reaction of hydroxyphtalimide with the halogeno derivatives or the corresponding alcohols using Mitsunobu reaction conditions. The affinity of the compounds for chicken brain melatonin receptors and recombinant human MT(1) and MT(2) receptors was evaluated using 2-[125I] iodomelatonin as the radioligand. Overall, the introduction of an oxygen atom in the amido chain was not a favourable parameter as the compounds were less potent than the corresponding deoxy derivatives. However, nanomolar compounds were obtained with the arylethyloxy derivatives (13c (R'=nPr), chicken brain, hMT(1), hMT(2), K(i) values: 4.8, 3.86, 2.4 nM, respectively) and the 2,7 dimethoxynaphthalene derivatives (17c (R'=nPr), chicken brain, hMT(1), hMT(2), K(i) values: 0.04, 0.13, 0.1 nM, respectively). The functional activity of these compounds was evaluated by the aggregation of melanophores in Xenopus laevis tadpoles and the potency was related to the affinity of the molecules for melatonin receptors. The compounds were found to be full agonists and compound 17a was 20-fold more potent than melatonin in this bioassay. PMID- 12538012 TI - Inhibitory effects of lapachol derivatives on epstein-barr virus activation. AB - Sixteen derivatives (2-17) synthesized from the naphthoquinone lapachol (1), were tested for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O-tetradecanoylphorbol-13-acetate (TPA), as a test for potential cancer chemopreventive agents. They exhibited a variety of inhibitory activities from very high to moderate, which allow us to suggest structure activity relationships. Ten of these derivatives are reported for the first time, their structures being thoroughly determined by spectroscopic methods. PMID- 12538013 TI - Synthesis and insecticidal activity of novel dihydropyrrole derivatives with N sulfanyl, sulfinyl, and sulfonyl moieties. AB - This paper reports the synthesis and insecticidal activity of a new type of dihydropyrrole derivatives with sulfur moieties such as sulfanyl, sulfinyl, and sulfonyl groups at the 1-position. These derivatives exhibited high insecticidal potency against Nilaparvata lugens and Nephotettix cincticeps. Investigation of the structure-activity relationships revealed that the alkoxycarbonyloxy groups at the 4-position tended to increase the systemic insecticidal activity. PMID- 12538014 TI - Synthesis and in vitro antitumoral activity of new N-phenyl-3 pyrrolecarbothioamides. AB - A new series of N-phenylpyrrolecarbothioamides were obtained from base catalyzed intramolecular cyclization of 3-amino-3-(alkyl or arylamino)propenethioamides. Pyrrole derivatives were evaluated for their in vitro anticancer activity toward cell lines of nine different types of human cancer. Some of newly prepared compounds demonstrated inhibitory effects on the growth of a wide range of cancer cell lines generally at 10(-6) M level and in some case at 10(-8) M concentrations. PMID- 12538015 TI - Mode of binding of the cytotoxic alkaloid berberine with the double helix oligonucleotide d(AAGAATTCTT)(2). AB - Berberine, an isoquinoline plant alkaloid, belongs to the structural class of protoberberines. Recently, the ability of these compounds to act as Topoisomerase I or II poisons, was related to the antitumor activity. The binding of protoberberins to DNA has been studied and the partial intercalation into the double helix has been considered responsible for their activity. We have studied the interaction of berberine with the double helix oligonucleotides d(AAGAATTCTT)(2), d(GCGATCGC)(2), d(CGTATACG)(2), d(CGTACG)(2), 5' d(ACCTTTTTGATGT)-3'/5(ACATCAAAAAGGT)-3' and with the single strand 5' d(ACATCAAAAAGGT)-3', by 1H, 31P NMR and UV spectroscopy. Phosphorus resonance experiments were performed to detect small conformational changes of the phosphoribose backbone, in the case that an intercalation process occurs. Our data reveal that berberine does not intercalate into the duplexes studied, and binds preferentially to AT rich sequences. The structure of the complex with d(AAGAATTCTT)(2) was determined by using proton 2D NOESY spectra, which allowed to obtain several NOE contacts between the drug and the nucleotide. Structural models were built up by Molecular Mechanics (MM) and Molecular Dynamics (MD) calculations, by using the inter-proton distances derived from the NOE values. Berberine results to be located in the minor groove, lying with the convex side on the helix groove and presenting the positively charged nitrogen atom close to the negative ionic surface of the oligomer. The large 1H chemical shifts variation, observed for the drug when it is added to the above duplexes, as well as to the single strand oligomer, was interpreted with non-specific ionic interactions. The binding constants were measured by UV and NMR spectroscopy. They are strongly affected by the ionic strength and by the self-association process, which commonly occurs with this type of drugs. A dimerisation constant was measured and the value was included in the calculations of the binding constants. The results obtained show that the non-specific ionic interactions represent the major contribution to the values of the binding constants. These parameters, as well as the protons chemical shift variation of the ligand, are thus not diagnostic for the identification of a drug/DNA complex. PMID- 12538016 TI - Importance of the thiomorpholine introduction in new pyrrole derivatives as antimycobacterial agents analogues of BM 212. AB - During the course of our investigations in the field of azole antimicrobial agents, we have identified BM 212, a pyrrole derivative with good in vitro activity against mycobacteria and candidae. These findings prompted us to prepare new pyrrole derivatives 1-10 in the hope of increasing the activity. The microbiological data showed interesting in vitro activity against Mycobacterium tuberculosis and atypical mycobacteria. PMID- 12538017 TI - 2,3-Dimethoxybenzo[i]phenanthridines: topoisomerase I-targeting anticancer agents. AB - Appropriately substituted benzo[i]phenanthridines structurally related to nitidine, a benzo[c]phenanthridine alkaloid with antitumor activity, are active as topoisomerase I-targeting agents. Studies on benzo[i]phenanthridines have indicated analogues that possess a 2,3-methylenedioxy moiety and at least one and preferably two methoxyl groups at the 8- and 9-positions, such as 8,9-dimethoxy 2,3-methylenedioxybenzo[i]phenanthridine, 2, are active as topoisomerase I targeting agents. Tetramethoxylated benzo[i]phenanthridines, wherein the 2,3 methylenedioxy moiety is replaced with methoxyl groups at the 2- and 3-position, are inactive as a topoisomerase I-targeting agent. These results initially suggested that the 2,3-methylenedioxy moiety was critical to the retention of potent activity. Further studies revealed that 2,3-dimethoxy-8,9 methylenedioxybenzo[i]phenanthridine, 7a, is more potent than 2 as a topoisomerase I-targeting agent. The observation that 2,3-dimethoxylated benzo[i]phenanthridines can actually exhibit enhanced activity prompted the present study in which several 8-substituted 2,3-dimethoxybenzo[i]phenanthridines were prepared and their pharmacological activities evaluated. The influence of NH(2), CN, CH(2)OH, OBn, OCH(3), OH, and NHCOCH(3 )substituents at the 8-position on the relative activity of these 2,3-dimethoxybenzo[i]phenanthridines was examined. Relative to these derivatives, 7a was the most potent topoisomerase I targeting agent, possessing similar cytotoxicity to that of nitidine in the human lymphoblast tumor cell line, RPMI8402. PMID- 12538018 TI - Synthetic and novel biocatalytic resolution studies on (+/-)-5/6/7-acetoxy-4-aryl 3,4-dihydrocoumarins. AB - Eleven (+/-)-5/6/7-acetoxy-4-aryl-3,4-dihydrocoumarins have been synthesised in two steps starting from the coupling of cinnamic acid/substituted cinnamic acid with appropriate phenols, followed by acetylation in 50-83% overall yields. All hydroxy- and acetoxycoumarins were unambiguously identified on the basis of their spectral data. Candida antarctica lipase-catalysed deacetylation of these racemic acetoxydihydrocoumarins in dioxane occurred with moderate enantioselectivity. This is one of the rare examples of resolution using phenolic ester moiety as a remote handle for chiral recognition by a lipase. PMID- 12538019 TI - 3-Indolyl-1-naphthylmethanes: new cannabimimetic indoles provide evidence for aromatic stacking interactions with the CB(1) cannabinoid receptor. AB - A series of 1-pentyl-1H-indol-3-yl-(1-naphthyl)methanes (9-11) and 2-methyl-1 pentyl-1H-indol-3-yl-(1-naphthyl)methanes (12-14) have been synthesized to investigate the hypothesis that cannabimimetic 3-(1-naphthoyl)indoles interact with the CB(1) receptor by hydrogen bonding to the carbonyl group. Indoles 9-11 have significant (K(i)=17-23nM) receptor affinity, somewhat less than that of the corresponding naphthoylindoles (5, 15, 16). 2-Methyl-1-indoles 12-14 have little affinity for the CB(1) receptor, in contrast to 2-methyl-3-(1-naphthoyl)indoles 17-19, which have affinities comparable to those of 5, 15, 16. A cannabimimetic indene hydrocarbon (26) was synthesized and found to have K(i)=26+/-4nM. Molecular modeling and receptor docking studies of naphthoylindole 16, its 2 methyl congener (19) and indolyl-1-naphthylmethanes 11 and 14, combined with the receptor affinities of these cannabimimetic indoles, strongly suggest that these cannabinoid receptor ligands bind primarily by aromatic stacking interactions in the transmembrane helix 3-4-5-6 region of the CB(1) receptor. PMID- 12538020 TI - The structural basis for kainoid selectivity at AMPA receptors revealed by low mode docking calculations. AB - The kainoids are a class of excitatory and excitotoxic pyrrolidine dicarboxylates that act at ionotropic glutamate receptors. The kainoids bind kainate receptors with high affinity and, while binding affinity is lower at AMPA (alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid) receptors, they are active in functional assays at this receptor subtype as well. However, kainoids are only partial agonists at AMPA receptors. Currents evoked by kainoids have been described as either slowly desensitizing, partially desensitizing, or non desensitizing. Recently acquired X-ray crystal structures of the ligand binding domain of the iGluR2, AMPA sensitive receptor suggest that differences in ligand receptor interactions may influence functional properties of an agonist. In an effort to identify important ligand-receptor interactions of various kainoids, we have conducted a series of low-mode docking searches of AMPA agonists in the iGluR2 binding domain. Kainic acid exhibited alternate low-lying geometries, with loss of hydrogen bonds to domain 2, which may represent a dissociation route not available to other kainoids. The most potent of the kainoids are capable of forming hydrogen bonding interactions that span the two domains of the receptor. In particular, a hydrogen bond between the domoic acid C6' carboxylic acid and Ser652 may prevent a peptide bond rotation that is associated with the desensitized state of the receptor. PMID- 12538021 TI - Synthesis and biological evaluation of 2,7-Dihydro-3H-dibenzo[de,h]cinnoline-3,7 dione derivatives, a novel group of anticancer agents active on a multidrug resistant cell line. AB - A series of anthrapyridazone derivatives with one or two basic side chains at various positions in the tetracyclic chromophore have been synthesized. The key intermediates in the synthesis are 2,7-dihydro-3H-dibenzo[de,h]cinnoline-3,7 diones 1, 12 and 15 monosubstituted at position 2 (4d, 16a-e), or 6 (2a-f) or disubstituted at positions 2 and 6 (4a-c) or 2 and 8 (17a-e) with appropriate alkylaminoalkylamines. All analogues showed in vitro cytotoxic activity against murine leukemia (L1210) and human leukemia (K562) cell lines. The compounds were also active against human leukemia multidrug resistant (K562/DX) cell line with resistance index (RI) in the range 1-3 depending on the compound's structure. Two of the most active in vitro compounds 4a and 11 were tested in vivo against murine P388 leukemia and displayed antileukemic activity comparable with that of Mitoxantrone. DNA-binding assays were performed and DNA affinity data were correlated with the structures of the compounds. The cytoplasmatic membrane affinity values (log k'(IAM)) have also been determined and the correlation with the resistance indexes discussed. The anthrapyridazones constitute a novel group of antitumor compounds that can overcome multidrug resistance. PMID- 12538022 TI - Non-antibiotic antibacterial activity of dodecyl gallate. AB - Dodecyl (C(12)) gallate (3,4,5-trihydroxybenzoate) (1) was found to possess antibacterial activity specifically against Gram-positive bacteria, in addition to its potent antioxidant activity. The time-kill curve study indicates that this amphipathic gallate exhibits bactericidal activity against methicillin resistant Staphylococcus aureus (MRSA) strains. Dodecyl (lauryl) gallate inhibited oxygen consumption in whole cells and oxidation of NADH in membrane preparation. The antibacterial activity of this gallate comes in part from its ability to inhibit the membrane respiratory chain. As far as alkyl gallates are concerned, their antimicrobial spectra and potency depend in part on the hydrophobic portion of the molecule. PMID- 12538023 TI - Pyrazolinone-piperidine dipeptide growth hormone secretagogues (GHSs). Discovery of capromorelin. AB - Novel pyrazolinone-piperidine dipeptide derivatives were synthesized and evaluated as growth hormone secretagogues (GHSs). Two analogues, capromorelin (5, CP-424391-18, hGHS-R1a K(i)=7 nM, rat pituicyte EC(50)=3 nM) and the des-methyl analogue 5c (hGHS-R1a K(i)=17 nM, rat pituicyte EC(50)=3 nM), increased plasma GH levels in an anesthesized rat model, with ED(50) values less than 0.05 mg/kg iv. Capromorelin showed enhanced intestinal absorption in rodent models and exhibited superior pharmacokinetic properties, including high bioavailabilities in two animal species [F(rat)=65%, F(dog)=44%]. This short-duration GHS was orally active in canine models and was selected as a development candidate for the treatment of musculoskeletal frailty in elderly adults. PMID- 12538024 TI - Relationships between structure, antibacterial activity, serum stability, pharmacokinetics and efficacy in 3-(heteroarylthio)cephems. Discovery of RWJ 333441 (MC-04,546). AB - SAR studies in a series of related 3-(heteroarylthio)cephems determined that a relatively high chemical reactivity of the beta-lactam ring, modulated by electronic effects of substituents at C-3 and C-7, is necessary to achieve high in vitro activity against methicillin-resistant Staphylococcus aureus (MRSA). Such high reactivity results in lowered hydrolytic stability and concomitantly increases susceptibility to beta-lactam ring opening mediated by serum enzymes. Therefore, optimization of anti-MRSA activity versus stability toward serum mediated degradation required a fine balance of substituent effects. Serum stability studies (measured as percentage of parent drug degraded after 60 min incubation) revealed up to 80-fold difference in degradation rate in a series of closely related (3-heteroarylthio)cephems. Of the compounds evaluated, RWJ-333441 (MC-04,546) possessed the best balance of serum stability (6% degradation after 60 min incubation) and in vitro activity versus MRSA (S. aureus COL MIC=1 microgram/mL). Accordingly, RWJ-333441 displayed excellent in vivo efficacy versus methicillin-susceptible Staphylococcus aureus (MSSA, ED(50)=0.39 mg/kg in mouse sepsis model with S. aureus Smith) and good pharmacokinetic properties in the rat (Cl(total)=0.39 L/h/kg). PMID- 12538025 TI - Inhibition and kinetics of mycobacterium tuberculosis and mycobacterium smegmatis mycothiol-S-conjugate amidase by natural product inhibitors. AB - The current rise in mycobacterial-related infections and disease, coupled with drug resistance, underlines the continuing need for new antimycobacterials. To this end, we have screened approximately 1500 extracts derived from marine plants and invertebrates and terrestrial fungi for their ability to inhibit a newly described mycobacterial detoxification enzyme mycothiol-S-conjugate amidase (MCA). As described in this paper, our screening and chemistry efforts thus far have led to the identification of 13 natural product inhibitors that represent six different structural classes. By conducting enzyme inhibition assays using varied inhibitor and substrate concentrations, we have determined the mode of inhibition of Mycobacterium tuberculosis MCA for four of these compounds. We show that two types of bromotyrosine-derived natural products are competitive inhibitors of MCA; while oceanapiside, an alpha,omega-bis-aminohydroxy glycosphingolipid, and the fungal metabolite gliotoxin, a dithiadiketopiperazine, are simple and mixed non-competitive inhibitors, respectively. Correlation of these results with the chemical structures suggests that MCA is a metalloenzyme and that the oximinoamide and spiro-isoxazoline amide groups present in the competitive inhibitors are substrate mimics. PMID- 12538026 TI - Structure-activity relationships of 6-fluoroquinazolines: dual-acting compounds with inhibitory activities toward both TNF-alpha production and T cell proliferation. AB - We synthesized various 6-fluoro-7-(1-piperazino)quinazolines based on the structure of 1 and evaluated their inhibitory activities toward both TNF-alpha production and T cell proliferation responses. Among these compounds, 7a, having the 3,4-(methylenedioxy)phenyl moiety at the C(4)-position of the quinazoline ring, showed both inhibitory activities. Furthermore, the oral treatment with 7a exhibited an anti-inflammatory effect in rats with adjuvant arthritis as well as an inhibitory activity toward LPS-induced TNF-alpha production. PMID- 12538027 TI - Quantitative structure-activity relationships of phenolic compounds causing apoptosis. AB - A study of a variety of phenolic compounds (simple phenols, estradiol, bisphenol A, diethylstilbesterol) on their action on L1210 leukemia cells led to the formulation of the following QSAR for apoptosis:log 1/C=-3.16 Clog P+2.77 CMR 3.76n=11, r(2)=0.939, s=0.630, q(2)=0.892C is the molar concentration causing 25% apoptosis, Clog P is the calculated octanol/water partition coefficient and CMR is the calculated molecular refractivity. Our results imply the significance of characterization of the phenolic compounds with apoptotic activity and the development of new agents for cancer therapy. PMID- 12538029 TI - Estrogenic diazenes: heterocyclic non-steroidal estrogens of unusual structure with selectivity for estrogen receptor subtypes. AB - Estrogens regulate many biological functions, often acting in a tissue-selective manner. Their tissue-selective action is believed to involve differential estrogen action through the two estrogen receptor (ER) subtypes, ERalpha and ERbeta, as well as differential interaction of the ligand-receptor complexes with promoters and coregulator proteins. In the latter case, selectivity is based on the induction of specific conformations of the ligand-ER complex, conformations that are influenced by the structure of the ligand. Estrogen pharmaceuticals having an ideal balance of tissue-selective activity are being sought for menopausal hormone replacement, breast cancer prevention and therapy, and other actions. To expand on the structural diversity of ER ligands that might show such tissue selectivity, we have prepared a series of diazenes (pyrazines, pyrimidines, and pyridazines) substituted with two to four aryl groups and various short-chain aliphatic substituents. All of the pyrazine and pyrimidines bind to ER, some with high affinity and with a considerable degree of preferential binding to either ERalpha or ERbeta. One pyrimidine and one pyrazine have ERalpha affinity preferences as high as 23 and 9, respectively, and one pyrimidine has an ERbeta affinity preference of 8. The pyridazines, by contrast, are quite polar and have only very low binding affinity for the ER. In cell-based transcription assays, several of the pyrimidines and a pyrazine were found to be considerably more agonistic on ERalpha than on ERbeta. Because these triaryl diazenes have the largest volumes among the ER ligands so far investigated, their high affinity demonstrates the flexibility of the ligand binding pocket of the ERs and its tolerance for large substituents. Thus, these novel heterocyclic ligands expand the repertoire of chemical structures that bind to the estrogen receptor, and they could prove to be useful in elucidating the biological behavior of the two ER subtypes and in forming the basis for new estrogen pharmaceuticals having desirable tissue selectivity. PMID- 12538028 TI - Searching for allosteric effects via QSAR. Part II. AB - Allosteric interactions have in the past been established by means of X-ray crystallography or careful study of a single molecule at a variety of concentrations. Here we report a method for using QSAR to establish a change in reaction mechanism by establishing an inversion point. That is, as polarizability of a member of a congeneric set of compounds is increased (as measured by CMR), activity at first decreases until, at the inversion, activity turns around and increases. Out of 23 examples, 14 have inversion points of 10+/-1. This includes a wide variety of receptors such as thrombin, 5-HT, dopamine, and tyrosine kinase acting with a variety of ligands. PMID- 12538030 TI - Characterization of glucocorticoid receptor translocation, cytoplasmic IkappaB, nuclear NFkappaB, and activation of NFkappaB in T lymphocytes exposed to stress inducible concentrations of corticosterone in vivo. AB - The present study was conducted to determine if selected events in glucocorticoid receptor (GR) signaling that have been identified using mostly in vitro approaches with synthetic glucocorticoids also occur in mature T cells exposed to relevant levels of corticosterone in vivo. In contrast to effects reported in vitro, corticosterone did not cause significant translocation of GR to the nucleus in splenic T cells, though it did increase the amount of nuclear GR in these cells capable of binding to a glucocorticoid response element. At most time points and dosages, corticosterone caused little or no change in cytoplasmic IkappaB or nuclear NFkappaB levels. Activation of T cells by anti-CD3 increased the amount of NFkappaB in the nucleus and decreased the amount of IkappaB in the cytoplasm. Corticosterone did not significantly inhibit the decrease in cytoplasmic IkappaB, but it did slightly diminish the increase in nuclear NFkappaB. The same dosages of corticosterone substantially suppressed anti-CD3 induced cytokine gene expression, indicating that a meaningful amount of glucocorticoid-mediated signaling (of some type) occurred in this experimental system. Thus, GR translocation per se seems not to be critical for GR-mediated signaling in vivo. Other considerations, such as the make-up of GR-containing complexes in the nucleus or unexpected sensitivity to small changes in total nuclear GR, may be important. Additionally, upregulation of IkappaB and consequent inhibition of NFkappaB activation are not prominent in vivo, in contrast to results obtained in vitro or with a synthetic glucocorticoid. PMID- 12538031 TI - Linomide inhibits insulitis and modulates cytokine production in pancreatic islets in the nonobese diabetic mouse. AB - Linomide is an immunomodulator which has been shown to potently inhibit autoimmunity in several animal models for human autoimmune diseases, including type I diabetes in the nonobese diabetic (NOD) mouse. In this study, we investigate the basis for Linomide's protective effects in the NOD mouse by immunohistochemical and RT-PCR analysis of the phenotype and cytokine expression by cells infiltrating the islets of Langerhans in the pancreas. Linomide treatment was found to reduce the infiltration of T cells, B cells, dendritic cells (DC) and MHC class II(+) cells into the islets, but did not reduce macrophage (MPhi) infiltration. This was seen following Linomide treatment at 3 5, 4-8 and 14-24 weeks of age and thus appears to be independent of the stage of the autoreactive process and the extent of insulitis. The reduced insulitis may be due to reduced expression of adhesion molecules since decreased numbers of islet-associated blood vessels expressing CD106 and MAdCAM-1 were detected following Linomide treatment. Furthermore, short term Linomide treatment (3 or 7 days), which did not alter the number of infiltrating cells, was found to inhibit the production of TNF-alpha which is known to induce the expression of CD106 and MAdCAM-1. These results suggest that the reduced insulitis observed in Linomide treated animals is secondary to a functional modulation of infiltrating cells. PMID- 12538032 TI - Mycophenolic acid inhibits inosine 5'-monophosphate dehydrogenase and suppresses immunoglobulin and cytokine production of B cells. AB - Mycophenolic acid (MPA) reversibly inhibits inosine 5'-monophosphate dehydrogenase (IMPDH), an enzyme involved in the de novo synthesis of guanine nucleotides. Previously, mycophenolate mofetil (MMF), the pro-drug of MPA, was shown to exert beneficial effects on the systemic lupus erythematosus (SLE)-like disease in MRLlpr/lpr mice. In this study, MPA's immunomodulating effects in vitro on the B cell hybridoma MAR 18.5 were investigated. The cells were exposed for MPA at either 1 or 10 microM for 24 h, and the levels of immunoglobulins, cytokines and lactate dehydrogensase in supernatants were measured. The frequency of immunoglobulin producing cells and the proliferation and viability of the cells was also investigated. MPA exposure reduced the frequency of immunoglobulin producing cells, decreased the levels of immunoglobulins and cytokines in the supernatants, and decreased the cell proliferation. MPA was slightly cytotoxic as indicated by increased lactate dehydrogenase (LDH) levels and reduced viability. All MPA-induced effects were totally reversed by the addition of guanosine to the cultures. Thus, since activated B lymphocytes play a central role in lupus and our results show that B cells are targets for MPA, we propose that direct effects on B cells may be an important mechanism for the ameliorating effects of MMF in SLE. PMID- 12538033 TI - Maturation-dependent expression of C1q binding proteins on the cell surface of human monocyte-derived dendritic cells. AB - The expression and cell surface levels of many important receptors are dependent on the maturation stage of dendritic cells (DCs), and related to the unique function of immature and mature DCs. In this report, we show, for the first time, that human monocyte-derived DCs express two types of C1q receptors, gC1qR and cC1qR. Furthermore, immature DCs secrete detectable amount of C1q into the culture supernatant. Immature DCs express higher cell surface levels of both C1qRs than mature ones, while the total C1qR protein and mRNA levels remain the same. The following experimental evidence support this conclusion: (1) Inflammatory cytokines and LPS, which induce maturation of DCs, downregulate surface expression of both C1qR molecules. (2) Cytokines and drugs (IL-10, IFN alpha, Dexamethasone), which keep DCs phenotypically and functionally immature, significantly upregulate the cell surface expression of both C1qRs. (3) Neither of these treatments changed the intracellular gC1qR level nor the gC1qR mRNA levels measured by real time RT-PCR. The elevated surface expression of C1qRs on DCs has been found to be not due to increased apoptosis or cell death as the result of DC treatment. Taken together, these data show that human monocyte derived DCs express gC1qR and cC1qR, their expression on the cell surface is maturation dependent, and immature DCs secrete C1q. These data strongly suggest the role of C1qRs in immature DC function and in the regulation of immune processes. PMID- 12538034 TI - Anticellular and immunosuppressive properties of ethanolic extract of Acorus calamus rhizome. AB - Modulation of immune response to alleviate disease has been of interest since long. Plant extracts have been widely investigated for possible immunomodulatory properties. We have evaluated the anticellular and immunomodulatory properties of ethanolic extract of Acorus calamus rhizome. This extract inhibited proliferation of mitogen (phytohaemagglutinin; PHA) and antigen (purified protein derivative; PPD)-stimulated human peripheral blood mononuclear cells (PBMCs). In addition, A. calamus extract inhibited growth of several cell lines of mouse and human origin. It also inhibited production of nitric oxide (NO), interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-alpha). Intracytoplasmic interferon-gamma (IFN-gamma) and expression of cell surface markers, CD16 and HLA-DR, on human PBMC, were not affected on treatment with A. calamus extract but CD25 expression was down regulated. Our study demonstrates the antiproliferative and immunosuppressive potential of ethanolic extract of A. calamus rhizome in vitro. PMID- 12538035 TI - Chemically modified tetracyclines induce cytotoxic effects against J774 tumour cell line by activating the apoptotic pathway. AB - Here, we have studied the effects of chemically modified tetracyclines (CMTs) on apoptosis both at the level of the cytoplasmic proteolytic caspase cascade, and on Bcl-2 and c-myc mRNA expression in the J774 macrophage cell line. The results indicate that CMTs induce morphological changes consistent with apoptotic events, as clearly demonstrated both by the acridine orange and ethidium bromide staining, and by TUNEL and fragmentation ELISA assays. Furthermore, the analysis of the cell cycle by flow cytometry shows an evident apoptotic sub-G0G1 peak, without important modifications in the cell cycle distribution. CMTs induce programmed cell death (PCD) in a dose-dependent manner and CMT-8 is the strongest among them. CMT-1 and CMT-8 activate mainly caspase-8 as attested by the inhibitory effects of Z-VAD-fmk and Z-IEDT-fmk on CMT-induced apoptosis. Part of CMT-induced PCD is due to the activation of caspase-9, since it is reduced by the specific caspase-9 inhibitor, Z-LEHD-fmk. Besides, CMTs increase Bcl-2 and c-myc mRNA expression. Collectively, these data indicate that CMTs are potentially anti tumour agents, since they strongly trigger apoptosis both activating the proteolytic system of the caspase family and modulating genes involved in PCD regulation. PMID- 12538036 TI - Effect of Ginkgo biloba extract, EGb 761, on the cellular immune response in a hypothalamic-pituitary-adrenal axis activation model in the rat. AB - We evaluated the effects of Ginkgo biloba extract (EGb 761) on the cellular immune response of rats with immunosuppression induced by activation of the hypothalamic-pituitary-adrenal (HPA) axis. Groups of five rats were subjected to chronic stress by the application of daily electric shocks (ES) over 7 days. This stress produced a significant decrement in the delayed-type hypersensitivity response (DTH) to dinitrofluorobenzene (DNFB), and a decrease in the proliferation index of splenocytes. Treatment with oral doses of the phytopharmaceutical EGb 761 (100 mg/kg per day over 7 days) restored both the DTH response to DNFB and the proliferation index. EGb 761 has stress-alleviating properties through its moderation of corticosterone levels. It also possesses antioxidant activity that may contribute to its effects on the immune response. Our observations indicate that the phytopharmaceutical EGb 761 possesses immunostimulatory properties. PMID- 12538037 TI - Serum concentrations of growth factors in women with and without endometriosis: the action of anti-endometriosis medicines. AB - Endometriosis is a common gynecologic syndrome of unknown etiology and pathogenesis. Growth factors and inflammatory mediators produced by peritoneal leukocytes have recently been postulated to participate in the pathogenesis of endometriosis. Angiogenic factors released from peritoneal macrophages may also play a role in the development of this disease. In the present study, we investigate the soluble levels of vascular endothelial growth factor (VEGF), epidermal growth factor-receptor (EGF-R), granulocyte/macrophage-colony stimulating factor (GM-CSF), Insulin-like growth factor-1 (IGF-1) and interferon gamma (IFN-gamma) in the serum of 28 women with and 20 without endometriosis. We also compared these levels before, during and after treatment with danazol and leuprorelin acetate depot, the two therapeutic regiments of choice concerning this disease. We found that only sVEGF levels were higher in women with endometriosis in comparison to controls (P < 0.001) while sEGF-R is not present. GM-CSF, IGF-1 and IFN-gamma soluble levels are not affected in either healthy or endometriotic subjects. The 6-month treatment with danazol decreased sVEGF levels (P < 0.02) and increased sEGF-R levels (P < 0.001). These observations support the view that VEGF may be associated with the disease process and that danazol may bring sVEGF levels to a normal threshold. However, future studies will be focused on the anti-angiogenic control of the action of VEGF in patients with endometriosis. PMID- 12538038 TI - Influence of the thr164ile polymorphism in the beta2-adrenoceptor on the effects of beta-adrenoceptor agonists on human lung mast cells. AB - We have examined the influence of the thr164ile polymorphism in the beta(2) adrenoceptor on the ability of the beta-adrenoceptor agonists, isoprenaline and salbutamol, to stabilise human lung mast cells. A total of 124 mast cell preparations were genotyped and, of these, 120 were found to be homozygous (thr164thr) at position 164 of the beta(2)-adrenoceptor and 4 were heterozygous (thr164ile). None of the preparations was homozygous for ile at position 164. In these preparations, the effects of isoprenaline and salbutamol on the IgE mediated release of histamine from mast cells were studied. Both isoprenaline and salbutamol inhibited histamine release in a concentration-dependent manner. Average inhibitory potencies for both agonists, as assessed by pD(2) values, were higher in homozygous than in heterozygous preparations. For isoprenaline, this difference was statistically significant (P < 0.005), whereas for salbutamol, it was not (P = 0.21). These data suggest that the thr164ile polymorphism in the beta(2)-adrenoceptor may influence the extent to which certain beta-adrenoceptor agonists inhibit the responses of mast cells. PMID- 12538039 TI - Regulation of macrophage function by the antioxidant N-acetylcysteine in mouse oxidative stress by endotoxin. AB - Changes in several functions of peritoneal macrophages from mice with oxidative stress caused by intraperitoneal injection of endotoxin (Escherichia coli lipopolysaccharide, LPS) (100 mg/kg), and associated with a high production of reactive oxygen species (ROS), have been observed in our previous studies. Antioxidants such as N-acetylcysteine (NAC) are free radical scavengers that improve and modulate the immune response, especially in oxidative stress situations. Therefore, in the present work, we have studied the effects of the administration of NAC (150 mg/kg i.p.) on different functions of peritoneal macrophages from Swiss mice suffering that oxidative stress, caused by LPS (100 mg/kg). NAC was injected 30 min after LPS injection, and the peritoneal macrophages were obtained at 2, 4, 12, and 24 h after endotoxin injection. The following functions, key stages of the phagocytic process, were studied: adherence to substrate, chemotaxis, ingestion of particles, and production of ROS (reactive oxygen species), as well as tumor necrosis factor (TNFalpha) release. The decrease in chemotaxis and the increase in adherence, ingestion, superoxide anion production, and TNFalpha release shown by macrophages from animals with oxidative stress were counteracted by NAC injection. These data suggest that NAC administration may be useful for the treatment of oxidative stress-linked endotoxic shock, modulating the function of macrophages, specifically in decreasing the production of ROS and of inflammatory cytokines such as TNFalpha. PMID- 12538040 TI - In vivo thymulin treatments enhance avian lung natural killer cell cytotoxicity in response to infectious bronchitis virus. AB - Previous work has shown that in vitro thymulin treatments have the ability to enhance natural killer (NK) cell cytotoxicity. The purpose of the experiments presented here was to examine the in vivo effects of thymulin on avian NK cell activity in response to a viral infection. Five and a half-week-old K-strain chickens infected with avian infectious bronchitis virus (IBV) served as the model for these experiments. Daily thymulin injections began at varying time points prior to or post-infection. The controls received daily injections of the ZnCl(2)-containing carboxymethyl-cellulose (CMC) diluent. A 51Cr-cytolytic release assay was used to determine the activity of the NK cells harvested via lung lavage from the respiratory tracts of infected chickens. The results of these experiments showed that in vivo thymulin treatments enhance NK cytotoxicity. The greatest enhancement of NK cytotoxicity was observed at 10 days post-infection in those chickens that began receiving thymulin after infection. These results suggest that thymulin may not only have a role in enhancing immunosurveillance but also in enhancing the response of the innate immune system following infection. Dose-response experiments found that the 50 ng/100 g body weight (Bwt) dose significantly depressed the cytolytic activity of the NK cells in comparison to either the 10 ng/100 g Bwt dose or the control. PMID- 12538041 TI - Freund adjuvant induces TLR2 but not TLR4 expression in the liver of mice. AB - Freund adjuvants are used extensively to establish experimental animal models of autoimmune diseases and to produce antibodies. However, studies on their mechanisms of action have been largely neglected, particularly their effects on liver, the primary target organ for host-microbe interaction. Here we show that treatment with either complete (CFA) or incomplete (IFA) Freund adjuvant induced a 5-10-fold increase in toll-like receptor (TLR) 2 mRNA but not TLR4 mRNA in livers of mice. Since CFA is essentially made of killed Mycobacterium tuberculosis bacilli (Mtb) dissolved in IFA, it is the solvent in CFA that induced an increase in TLR2 expression. As TLR2 is the receptor activated by killed Mtb, this solvent-mediated increase in TLR2 expression will result in enhanced recognition of killed Mtb by hepatocytes during CFA administration. We propose that the potency of Freund adjuvant in eliciting an immune response lies in their ability to induce expression of the appropriate TLR, TLR2, for the active ingredient, killed Mtb, in CFA. PMID- 12538042 TI - Innate recognition of lipopolysaccharide by CD14 and toll-like receptor 4-MD-2: unique roles for MD-2. AB - Adaptive immunity generally refers to the ability of lymphocytes to recognize microbial, viral and fungal proteins via T cell receptors and antibodies. More ancestral and widespread innate immune mechanisms include those responsible for recognition of microbial glycolipids. Lipopolysaccharide (LPS) is the best studied, and arguably one of the most important of bacterial products because of its role in innate immune responses and endotoxin-mediated sepsis. Converging studies in two independent fields have recently led to the identification of LPS recognition molecules utilized by mammalian cells. Toll-like receptor 4 (TLR4) was identified as a mammalian homologue of the Toll receptor, which recognized fungi in the Drosophila's immune system. Spontaneous and targeted mutations identified TLR4 as an LPS recognition molecule. Separate studies of a Radioprotective 105 (RP105) and MD-1 heterodimer expressed by cells led to the identification of MD-2 as a molecule associated with TLR4. Very recent in vivo studies have now revealed an essential contribution of MD-2 to LPS recognition. These findings further our understanding of protective, as well as detrimental innate immune mechanisms and may lead to new therapies for endotoxin shock. PMID- 12538043 TI - Anti-allergic action of buckwheat (Fagopyrum esculentum Moench) grain extract. AB - The anti-allergic action of buckwheat grain extract (BGE) was investigated using rodent experimental models. The oral, intraperitoneal and intradermal administration of BGE significantly inhibited the compound 48/80-induced vascular permeability documented by Evans blue extravasation. In addition, BGE showed potent inhibitory effect on passive cutaneous anaphylaxis (PCA) activated by anti dinitrophenyl (DNP) IgE when orally administered. In an in vitro study, BGE revealed to possess inhibitory potential on the compound 48/80-induced histamine release from rat peritoneal mast cells (RPMC). Moreover, BGE inhibited the IL-4 and TNF-alpha mRNA induction by PMA and A23187 in human leukemia mast cells, HMC 1. Taken together, these results suggest that anti-allergic action of BGE may be due to the inhibition of histamine release and cytokine gene expression in the mast cells. PMID- 12538044 TI - Immunization to nicotine with a peptide-based vaccine composed of a conformationally biased agonist of C5a as a molecular adjuvant. AB - This paper describes the synthesis of a nicotine hapten (Nic) that possesses a carboxyl sidearm functional group allowing for conjugation to a peptide via amide bond formation. Nic was attached to the N-terminal amino group of a 19-residue peptide composed of a conformationally biased agonist of human C5a (YSFKPMPLaR), which is used as a molecular adjuvant and a B cell epitope of human MUC1 glycoprotein (YKQGGFLGL) to yield a peptide-based nicotine vaccine, NicYKQGGFLGLYSFKPMPLaR. Rats immunized with this vaccine were significantly less sensitive to behavioral effects (a Pavlovian discrimination task) induced by their exposure to high concentrations of nicotine (0.4 mg/kg) relative to their non-vaccinated counterparts. The attenuation of these nicotine-induced behavioral effects emanated from the presence of nicotine-specific antibodies (Abs) that were present in the sera of vaccinated rats even after their repeated exposure to high concentrations of nicotine during the time required to perform the behavioral assays. These results suggest that immunization with NicYKQGGFLGLYSFKPMPLaR in the absence of adjuvant is an effective means of inducing a nicotine-specific Ab response, which is capable of attenuating nicotine-induced behavioral/psychoactive effects. PMID- 12538046 TI - A great talent--a merciless fate. PMID- 12538047 TI - An appreciation of William H. Orme-Johnson III. AB - This short memoir of personal and professional interactions with Bill Orme Johnson covers a period of more than 35 years. Included are reflections on the early history of copper oxidases, iron-sulfur proteins and nitrogenase. PMID- 12538048 TI - Photo-lability of CO bound to Mo-nitrogenase from Azotobacter vinelandii. AB - In the presence of CO and under turnover conditions, Mo-nitrogenase generates three different electron paramagnetic resonance (EPR) signals. One of the signals, lo-CO, is an S=1/2 signal and occurs under low CO concentrations. The other two signals, hi-CO (S=1/2) and hi(5)-CO (S=3/2) displace the lo-CO as the CO concentration is raised above 0.05 atm. Irradiation of hi-CO with visible light at 12 K converts it into lo-CO. Using a series of color filters, the corrected action spectrum is determined and shown to contain 2-3 broad maxima in the region 350-730 nm. The conversion of lo-CO back into hi-CO is accomplished by warming the sample to 77 K for 5 min. Using this temperature cycle, the rate constant for the re-association of CO with lo-CO to form hi-CO is determined in the range 12-90 K. From these data, the activation energy for this reaction is calculated to be 3.9 kJ/mol. Identical irradiation of either lo-CO or hi(5)-CO induces no spectral change, showing that both of these states are photo-stable. The photo-stability of hi(5)-CO demonstrates that it is structurally different from hi-CO. PMID- 12538049 TI - Effects of substrates (methyl isocyanide, C2H2) and inhibitor (CO) on resting state wild-type and NifV(-)Klebsiella pneumoniae MoFe proteins. AB - We report the use of electron nuclear double resonance (ENDOR) spectroscopy to examine how the metal sites in the FeMo-cofactor cluster of the resting nitrogenase MoFe protein respond to addition of the substrates acetylene and methyl isocyanide and the inhibitor carbon monoxide. 1H, 57Fe and 95Mo ENDOR measurements were performed on the wild-type and the NifV(-)proteins from Klebsiella pneumoniae. Among the molecules tested, only the addition of acetylene to either protein induced widespread changes in the 57Fe ENDOR spectra. Acetylene also induced increases in intensity from unresolved protons in the proton ENDOR spectra. Thus we conclude that acetylene may bind to the resting-state MoFe protein to perturb the FeMo-cofactor environment. On the other hand, the present results show that methyl isocyanide and carbon monoxide do not substantially alter the FeMo cofactor's geometric and electronic structures. We interpret this as lack of interaction between those two molecules and the FeMo cofactor in the resting state MoFe protein. Thus, although it is generally accepted that substrates or inhibitors bind to the FeMo-cofactor only under turnover condition, this work provides evidence that at least one substrate can perturb the active site of nitrogenase under non-catalytic conditions. PMID- 12538050 TI - Identification and preliminary characterization of AcsF, a putative Ni-insertase used in the biosynthesis of acetyl-CoA synthase from Clostridium thermoaceticum. AB - The acsABCDE genes in the Clostridium thermoaceticum genome are used for autotrophic acetyl-CoA synthesis using the Wood-Ljungdahl pathway. A 2.8-kb region between acsC and acsD was cloned and sequenced. Two open reading frames, orf7 (approximately 1.9 kb) and acsF (approximately 0.7 kb) were identified. orf7 appears to encode an Fe-S protein, in that it contains five conserved cysteine residues, three of which are present in a motif (CGGXXXCGXC) commonly used to coordinate Fe-S clusters. However, Orf7 is probably not involved in autotrophic acetyl-CoA synthesis, as homologous genes are present in organisms that do not utilize this pathway and are absent in many that do. In contrast, acsF is probably involved in this pathway. Sequence alignment of AcsF and eleven homologs reveals a number of conserved regions, including a P-loop that binds nucleoside triphosphates and catalyzes their hydrolysis. One homolog is CooC, an ATPase/GTPase that inserts Ni into a precursor form of the C-cluster of the carbon monoxide dehydrogenase (CODH) from Rhodospirillum rubrum. Purified AcsF lacked Ni and Fe, and slowly catalyzed the hydrolysis of ATP. Such similarities to CooC suggest that AcsF may function to insert Ni into a Ni-deficient form of the bifunctional acetyl-CoA synthase/CODH from C. thermoaceticum (ACS(Ct)). However, this could not be established, as expression of acsF did not effect activation of recombinant AcsAB expressed in E. coli. Also, E. coli cells defective in hypB retained the ability to synthesize active recombinant AcsAB. Rather, the concentration of extracellular Ni(2+) ions was critical to activation. PMID- 12538051 TI - Refinement of the nickel site structure in Desulfovibrio gigas hydrogenase using range-extended EXAFS spectroscopy. AB - We have reexamined the Ni EXAFS of oxidized, inactive (as-isolated) and H(2) reduced Desulfovibrio gigas hydrogenase. Better spatial resolution was achieved by analyzing the data over a 50% wider k-range than was previously available. A lower k(min) was obtained using the FEFF code for phase shifts and amplitudes. A higher k(max) was obtained by removing an interfering Cu signal from the raw spectra using multiple energy fluorescence detection. The larger k-range allowed us to better resolve the Ni-S bond lengths and to define more accurately the Ni-O and Ni-Fe bond lengths. We find that as-isolated, hydrogenase has two Ni-S bonds at approximately 2.2 A, but also 1-2 Ni-S bonds in the 2.35+/-0.05 A range. A Ni O interaction is evident at 1.91 A. The as-isolated Ni-Fe distance cannot be unambiguously determined. Upon H(2) reduction, two short Ni-S bonds persist at approximately 2.2 A, but the remaining Ni-S bonds lengthen to 2.47+/-0.05 A. Good simulations are obtained with a Ni-Fe distance at 2.52 A, in agreement with crystal structures of the reduced enzyme. Although not evident in the crystal structures, an improvement in the fit is obtained by inclusion of one Ni-O interaction at 2.03 A. Implications of these distances for the spin-state of H(2) reduced H(2)ase are discussed. PMID- 12538052 TI - Comparison of the reactivity of nitric oxide and nitroxyl with heme proteins. A chemical discussion of the differential biological effects of these redox related products of NOS. AB - Investigations on the biological effects of nitric oxide (NO) derived from nitric oxide synthase (NOS) have led to an explosion in biomedical research over the last decade. The chemistry of this diatomic radical is key to its biological effects. Recently, nitroxyl (HNO/NO(-)) has been proposed to be another important constituent of NO biology. However, these redox siblings often exhibit orthogonal behavior in physiological and cellular responses. We therefore explored the chemistry of NO and HNO with heme proteins in different redox states and observed that HNO favors reaction with ferric heme while NO favors ferrous, consistent with previous reports. Further results show that HNO and NO were equally effective in inhibiting cytochrome P450 activity, which involves ferric and ferrous complexes. The differential chemical behavior of NO and HNO toward heme proteins provides insight into mechanisms of activity that not only helps explain some of the opposing effects observed in NOS-mediated events, but offers a unique control mechanism for the biological action of NO. PMID- 12538053 TI - Metal ion catalyzed hydrolysis of ethyl p-nitrophenyl phosphate. AB - 15N isotope effects in the nitro group and 18O isotope effects in the phenolic oxygen have been measured for the hydrolysis of ethyl p-nitrophenyl phosphate catalyzed by several metal ions. Co(III)-cyclen at pH 7, 50 degrees C, gave an 15N isotope effect of 0.12% and an 18O one of 2.23%, showing that P-O cleavage is rate limiting and the bond is approximately 50% broken in the transition state. The active catalyst is a dimer and the substrate is presumably coordinated to the open site of one Co(III), and is attacked by hydroxide coordinated to the other Co(III). Co(III)-tacn under the same conditions shows a similar 15N isotope effect (0.13%), but a smaller 18O one (0.8%). Zn(II)-cyclen at pH 8.5, 80 degrees C, gave an 15N isotope effect of 0.05% and an 18O one of 0.95%, suggesting an earlier transition state. The catalyst in this case is monomeric, and thus the substrate is coordinated to one position and attacked by a cis-coordinated hydroxide. Eu(III) at pH 6.5, 50 degrees C, shows a very large 15N isotope effect of 0.34% and a 1.6% 18O isotope effect. The large 15N isotope effect argues for a late transition state or Eu(III) interaction with the nitro group, and was also seen in Eu(III)-catalyzed hydrolysis of p-nitrophenyl phosphate. PMID- 12538054 TI - Intrinsic tryptophan fluorescence as a probe of metal and alpha-ketoglutarate binding to TfdA, a mononuclear non-heme iron dioxygenase. AB - 2,4-Dichlorophenoxyacetic acid (2,4-D)/alpha-ketoglutarate (alphaKG) dioxygenase, TfdA, couples the oxidative decarboxylation of alphaKG to the oxidation of the herbicide 2,4-D using a mononuclear non-heme Fe(II) active site. The intrinsic tryptophan fluorescence associated with the four Trp residues in TfdA allows for the use of fluorescence spectroscopy to monitor the binding of iron and alphaKG to the enzyme. The fluorescence spectrum of TfdA is quenched by 50-85% upon addition of Fe(II) or alphaKG, allowing determination of their binding affinities (K(d)=7.45+/-0.61 and 3.35+/-0.35 microM, respectively). Cu, Zn, Mn, Co, Mg, and Ca dictations also quench the TfdA fluorescence with affinities similar to that of Fe(II), whereas monovalent cations such as Na, K, and Li do not. H114A and D116A mutant forms of TfdA, lacking either a histidine or aspartate metallocenter ligand, exhibit weaker affinity for both Fe(II) and alphaKG based on the fluorescence changes. Trp256 is predicted to lie within 5 A of the metal and alphaKG binding sites; however, its substitution by Phe or Leu has negligible effects on the Fe(II)- and alphaKG-dependent fluorescence quenching. Because Trp195 is predicted to be quite distant ( approximately 15 A) from the active site, we conclude that some combination of Trp113 and Trp248 serves as the reporter that senses metal and cofactor binding to TfdA. PMID- 12538055 TI - Proton-coupled electron transfer in Fe-superoxide dismutase and Mn-superoxide dismutase. AB - Fe-containing superoxide dismutase (FeSOD) and MnSOD are widely assumed to employ the same catalytic mechanism. However this has not been completely tested. In 1985, Bull and Fee showed that FeSOD took up a proton upon reduction [J. Am. Chem. Soc. 107 (1985) 3295]. We now demonstrate that MnSOD incorporates the same crucial coupling between electron transfer and proton transfer. The redox-coupled H(+) acceptor has been presumed to be the coordinated solvent molecule, in both FeSOD and MnSOD, however this is very difficult to test experimentally. We have now examined the most plausible alternative: that Tyr34 accepts a proton upon SOD reduction. We report specific incorporation of 13C in the C(zeta) positions of Tyr residues, assignment of the C(zeta) signal of Tyr34 in each of oxidized FeSOD and MnSOD, and direct NMR observations showing that in both cases, Tyr34 is in the neutral protonated state. Thus Tyr34 cannot accept a proton upon SOD reduction, and coordinated solvent is concluded to be the redox-coupled H(+) acceptor instead, in both FeSOD and MnSOD. We have also confirmed by direct 13C observation that the pK of 8.5 of reduced FeSOD corresponds to deprotonation of Tyr34. This work thus provides experimental proof of important commonalities between the detailed mechanisms of FeSOD and MnSOD. PMID- 12538056 TI - Crystal structure of substrate free form of glycerol dehydratase. AB - Glycerol dehydratase (GDH) and diol dehydratase (DDH) are highly homologous isofunctional enzymes that catalyze the elimination of water from glycerol and 1,2-propanediol (1,2-PD) to the corresponding aldehyde via a coenzyme B(12) dependent radical mechanism. The crystal structure of substrate free form of GDH in complex with cobalamin and K(+) has been determined at 2.5 A resolution. Its overall fold and the subunit assembly closely resemble those of DDH. Comparison of this structure and the DDH structure, available only in substrate bound form, shows the expected change of the coordination of the essential K(+) from hexacoordinate to heptacoordinate with the displacement of a single coordinated water by the substrate diol. In addition, there appears to be an increase in the rigidity of the K(+) coordination (as measured by lower B values) upon the binding of the substrate. Structural analysis of the locations of conserved residues among various GDH and DDH sequences has aided in identification of residues potentially important for substrate preference or specificity of protein protein interactions. PMID- 12538057 TI - Expression, purification and characterisation of a Bacillus subtilis ferredoxin: a potential electron transfer donor to cytochrome P450 BioI. AB - The fer gene from Bacillus subtilis has been subcloned and overexpressed in Escherichia coli and the protein (Fer) purified to homogeneity. N-Terminal sequencing and mass spectrometry indicate that the initiator methionine is removed from the protein and that the molecular mass is 8732 Da consistent with that deduced from the gene sequence. Amino-acid sequence comparisons indicate that Fer is a ferredoxin containing a 4Fe-4S cluster. The electron paramagnetic resonance spectrum of the reduced form of Fer is typical for a [4Fe-4S](+) cluster showing rhombic signals with g values of 2.07, 1.93 and 1.88. Reduced Fer also gives rise to a magnetic circular dichroism spectrum typical of a [4Fe 4S](+) cluster. Potentiometric titrations indicate that Fer has a reduction potential of -385+/-10 mV for the [4Fe-4S](+)-[4Fe-4S](2+) redox couple, well within the normal range expected for such a ferredoxin. A proposed physiological role for Fer is as an electron donor to cytochrome P450 BioI. Studies on Fer binding to P450 BioI give rise to a K(d) value of 0.87+/-0.10 microM. Anaerobic experiments using CO-saturated buffer indicate that Fer is indeed capable of transferring electrons to this cytochrome P450 albeit at a fairly low rate. PMID- 12538058 TI - NMR solution structures of two mutants of desulforedoxin. AB - The differences in geometry at the metal centres in the two known [Fe-4S] proteins rubredoxin (Rd) and desulforedoxin (Dx) are postulated to be a result of the different spacing of the C-terminal cysteine pair in the two proteins. In order to address this question, two mutants of Desulfovibrio gigas Dx with modified cysteinyl spacing were prepared and their solution structures have been determined by NMR. Mutant 1 of Dx (DxM1) has a single glycine inserted between the adjacent cysteines (C28 and C29) found in the wild type Dx sequence. Mutant 3 (DxM3) has two amino acid residues, -P-V-, inserted between C28 and C29 in order to mimic the primary sequence found in Rd from Desulfovibrio gigas. The solution structure of DxM1 exists, like wild type Dx, as a dimer in solution although the single glycine inserted between the adjacent cysteines disrupts the stability of the dimer resulting in exchange between a dimer state and a small population of another, probably monomeric, state. For DxM3 the two amino acid residues inserted between the adjacent cysteines results in a monomeric protein that has a global fold near the metal centre very similar to that found in Rd. PMID- 12538062 TI - A review of electrical impedance techniques for breast cancer detection. AB - Some evidence has been found that malignant breast tumors have lower electrical impedance than surrounding normal tissues. Although the separation of malignant tumors from benign lesions based on impedance measurements needs further investigation, electrical impedance could be used as an indicator for breast cancer detection. In this paper, we provide a systematic technical review of the existing electrical impedance techniques proposed for breast cancer detection, with an emphasis on noninvasive impedance imaging techniques. The electrical impedance of human breast tissue is first introduced, with tabulation of previous in vitro impedance measurement results on cancerous and normal breast tissues, and a brief description on the limited in vivo impedance measurements completed with invasive, or noninvasive, non-imaging techniques. A detailed review on noninvasive impedance imaging techniques for breast cancer detection, such as electrical impedance tomography (EIT) and electrical impedance mapping (EIM), is then presented. We suggest that for better breast cancer detection, an invasive impedance technique may be enhanced by combination with other cancer indicators. 3D EIT should be improved through collective efforts. EIM using a pair of electrode arrays is a viable method with great potential. Magnetic induction tomography and other magnetic induction based impedance imaging for breast cancer detection are promising and merit further exploration as well. PMID- 12538063 TI - Automated image analysis technique for measurement of femoral component subsidence in total hip joint replacement. AB - A new technique for the measurement of subsidence of the femoral components is proposed. The method relies on the implantation of two ball markers around the femoral stem. A single radiographic image of the hip is analysed using image processing techniques to minimize subjectivity related to manual identification of landmarks. Dimensions of the stem are used to correct for magnification and out-of-plane rotations resulting from radiographic positioning. This technique has been applied to a specific design of implant (Exeter). A study of the effect of radiographic positioning has been conducted using a cadaveric bone phantom. Results for the variation in the measurement of axial migration compared to the neutral position (in millimetres) were: 0.942 (10 degrees extension); 0.347 (20 degrees flexion); -0.435 (40 degrees internal rotation); 0.187 (30 degrees external rotation) for distances measured between the bone marker and the implant. Results for distances measured between the implant and the cement centralizer were: 0.107 (10 degrees extension); -0.277 (20 degrees flexion); 0.085 (40 degrees internal rotation); 0.280 (30 degrees external rotation). The variations from within a more realistic range of positions demonstrate that axial migration measurements of ca. 0.5 mm between the bone and implant, and less than 0.3 mm between the implant and the bone cement, may be expected. PMID- 12538064 TI - Finite element study of trochanteric gamma nail for trochanteric fracture. AB - A three-dimensional finite element study of trochanteric fracture fixation by a trochanteric gamma nail (TGN) was investigated in this study. The analyses were performed under one-legged stance load boundary conditions to study the stress distribution and displacements. The influence of material properties (E-modulus) of the implant, the bone and contact condition in the fracture zone was determined. The results show that the stresses in the implant were lower in case of titanium alloy implant material but at the same time higher displacements occurred. The results also indicate that the stresses in the TGN gradually reduced throughout the healing process of the bone in the fracture zone. PMID- 12538065 TI - Effects of condylar fibrocartilage on the biomechanical loading of the human temporomandibular joint in a three-dimensional, nonlinear finite element model. AB - The present study was undertaken to test a hypothesis that the addition of articular fibrocartilage in the condyle of the temporomandibular joint reduces three-dimensional stress distribution in the condyle, the disc and articular eminence. A three-dimensional, nonlinear finite-element model was developed for analysis of joint loading before and after the addition of condylar fibrocartilage to the osseous mandibular condyle reconstructed from spiral computer topography data. In the model, each of the disc, condyle and articular eminence was arbitrarily divided into five regions: the anterior, posterior, medial, lateral and central. Von Mises stresses that in virtually all regions of the disc, condyle and articular eminence became lower after the addition of condylar fibrocartilage. Especially remarkable was the approximately four-fold reduction in von Mises stresses in the anterior, central and medial regions of the mandibular condyle. In comparison, only slight to moderate stress reductions occurred in the disc and articular eminence, suggesting that condylar fibrocartilage absorbs considerable stresses and likely dampens more loads than the disc and articular eminence. The mandibular condyle demonstrated the largest total displacement in all directions after the addition of articular fibrocartilage, followed by the disc and articular eminence. We conclude that the addition of articular fibrocartilage primarily reduces loading of the mandibular condyle, rather than the disc and articular eminence. These findings lead to a hypothesis that the mandibular condyle more likely functions as a shock absorber than the disc. PMID- 12538066 TI - A theoretical study of chemical delivery within the lung using exogenous surfactant. AB - A mathematical model is developed for lung injury treatments involving the delivery of therapeutic chemicals, such as drugs and gene vectors, into the lung using simultaneous tracheal instillation of exogenous pulmonary surfactant. The influence of exogenous surfactant dose, flow rate, bulk liquid viscosity, pulmonary absorption rate and chemical molecular diffusivity on the chemical delivery to the lung is investigated. Our results reveal that different pulmonary absorption rates lead to significantly different distribution patterns and change the time taken for the total amount of chemical to be absorbed along the airways. The various factors can also influence where the majority of the chemical is placed within the lung and this is relevant to the targeting of drugs to particular lung generations. PMID- 12538067 TI - Quantitative assessment of co-contraction in cervical musculature. AB - The co-contraction of cervical musculature is quantitatively assessed. For the modeling procedure, electromyographic (EMG) signal and anatomical data of the neck musculature were used. EMG signals were collected from eight sites of the neck bilaterally using Ag-AgCl surface electrodes from ten adult male subjects. The subjects performed voluntary isometric contractions gradually developing to maximum efforts in flexion, extension, left lateral bending and right lateral bending. The EMG-assisted optimization modeling procedure was used to estimate muscle forces. To quantify co-contraction, muscle forces were decomposed into task subset and co-contraction subset of muscle forces. To show the degree of co contraction, 'co-contraction ratio' was defined as the proportion of co contraction muscle forces to total muscle forces. The ranges of co-contraction ratio are 0.08-0.16 during extension, 0.30-0.41 during flexion, 0.27-0.32 during left lateral bending, and 0.30-0.36 during right lateral bending. In all cases, co-contraction ratios increase as external moments increase. The ratios of compressive spinal loads from the co-contraction subset of muscle forces to those from total muscle forces are similar to co-contraction ratios during the whole ramp period. This study provides data demonstrating quantitative measures of the contribution of muscle co-contraction to cervical spinal loads. PMID- 12538068 TI - Evaluation of trajectories and contact pressures for the straight nucleus cochlear implant electrode array - a two-dimensional application of finite element analysis. AB - A two-dimensional (2D) finite element analysis has been used in this study to model the insertion of the Nucleus electrode array with different stiffness properties in order to evaluate the propensity of damage by visualizing the predicted trajectories and by comparing the buckling stresses and the contact pressures at the tip (and its distribution along the length) of the electrode array. Previous temporal bone studies have shown that damage during insertion of an electrode array around the basal turn of the cochlear spiral could be related to the design and the stiffness properties of the electrode array. However, it is difficult to evaluate different designs of electrode arrays purely by experimental methods as the experimental conditions and their results are difficult to reproduce. Three electrode arrays with different mechanical properties, i.e. uniform stiffness, graded stiffness, and a soft tip have been modelled. Buckling stress and contact pressure at the tip of the electrode array were found to be highest for the arrays with uniform stiffness. The contact pressures at the tip of the electrode array appeared strongly influenced by the stiffness profile and were optimal for graded stiffness. The results indicate the importance of the electrode array design and stiffness properties in minimizing trauma. However, there are a number of limitations in the present 2D evaluation which will require further analysis using a three-dimensional model to obtain definitive results. PMID- 12538069 TI - Automated cardiac MR image segmentation: theory and measurement evaluation. AB - We present a new approach to magnetic resonance image segmentation with a Gradient-Vector-Flow-based snake applied to selective smoothing filtered images. The system also allows automated image segmentation in the presence of grey scale inhomogeneity, as in cardiac Magnetic Resonance imaging. Removal of such inhomogeneities is a difficult task, but we proved that using non-linear anisotropic diffusion filtering, myocardium edges are selectively preserved. The approach allowed medical data to be automatically segmented in order to track not only endocardium, which is usually a less difficult task, but also epicardium in anatomic and perfusion studies with Magnetic Resonance. The method developed proceeds in three distinct phases: (a) an anisotropic diffusion filtering tool is used to reduce grey scale inhomogeneity and to selectively preserve edges; (b) a Gradient-Vector-Flow-based snake is applied on filtered images to allow capturing a snake from a long range and to move into concave boundary regions; and (c) an automatic procedure based on a snake is used to fit both endocardium and epicardium borders in a multiphase, multislice examination. A good agreement (P<0.001) between manual and automatic data analysis, based on the mean difference+/-SD, was assessed in a pool of 907 cardiac function and perfusion images. PMID- 12538071 TI - In vivo measurement of the elastic modulus of the human periodontal ligament. PMID- 12538072 TI - The evolution of desaturases. PMID- 12538073 TI - Recent advances in the study of fatty acid desaturases from animals and lower eukaryotes. AB - The biosynthesis of polyunsaturated fatty acids (PUFAs) in different organisms can involve a variety of pathways, catalyzed by a complex series of desaturation and elongation steps. A range of different desaturases have been identified to date, capable of introducing double bonds at various locations on the fatty acyl chain. Some recently identified novel desaturases include a delta4 desaturase from marine fungi, and a bi-functional delta5/delta6 desaturase from zebrafish. Using molecular genetics approaches, these desaturase genes have been isolated, identified, and expressed in variety of heterologous hosts. Results from these studies will help increase our understanding of the biochemistry of desaturases and the regulation of PUFA biosynthesis. This is of significance because PUFAs play critical roles in multiple aspects of membrane physiology and signaling mechanisms which impact human health and development. PMID- 12538074 TI - Mechanism of fatty acid desaturation: a bioorganic perspective. AB - The desaturation of long chain fatty acids is a ubiquitous transformation which plays a critical role in the biosynthesis of lipids. Of particular interest to the bioorganic chemist is the unique ability of desaturases to oxidize unactivated hydrocarbon chains in a chemo-, regio- and stereoselective manner. The mechanism of membrane-bound desaturases has been examined using regiospecifically labelled analogues bearing deuterium, sulfur or fluorine substituted methylene isosteres. These probes have been applied in the study of several biomedically important desaturase systems including a prototypical yeast stearoyl CoA delta(9) desaturase. In all cases, it has been found that the dehydrogenation (desaturation) process is initiated by a kinetically important hydrogen activation step at the carbon of the incipient double bond which is closest to the acyl terminus of the fatty acid chain. These results point to a common active site architecture which is highly conserved among a wide range of membranous desaturases. PMID- 12538075 TI - Role of stearoyl-coenzyme A desaturase in lipid metabolism. AB - Stearoyl-CoA desaturase (SCD) (EC 1.14.99.5) is an endoplasmic reticulum-bound enzyme that catalyzes the delta9-cis desaturation of saturated fatty acyl-CoAs, the preferred substrates being palmitoyl- and stearoyl-CoA, which are converted to palmitoleoyl- and oleoyl-CoA, respectively. These monounsaturated fatty acids are used as substrates for the synthesis of triglycerides, wax esters, cholesteryl esters and membrane phospholipids. The saturated to monounsaturated fatty acid ratio affects membrane phospholipid composition and alteration in this ratio has been implicated in a variety of disease states including cardiovascular disease, obesity, diabetes, neurological disease, skin disorders and cancer. Thus, the expression of SCD is of physiological importance in normal and disease states. Several mammalian SCD genes have been cloned. A single human, three mouse and two rat are the best characterized SCD genes. The physiological role of each SCD isoform and the reason for having three or more SCD gene isoforms in the rodent genome are currently unknown. A clue as to the physiological role of the SCD, at least SCD1 gene and its endogenous products came from recent studies of asebia mouse strains that have a natural mutation in the SCD1 gene and a mouse model with a targeted disruption of the SCD1 gene. In this review we discuss our current understanding of the physiological role of SCD in lipid synthesis and metabolism. PMID- 12538076 TI - Stearoyl-CoA desaturase, a short-lived protein of endoplasmic reticulum with multiple control mechanisms. AB - Stearoyl-CoA desaturase (SCD) is a short-lived, polytopic membrane-bound non-heme iron enzyme localized primarily in the endoplasmic reticulum. SCD is required for the biosynthesis of monounsaturated fatty acids, and plays a key role in hepatic synthesis of triglycerides and very-low-density lipoproteins. The intracellular concentration of SCD fluctuates in a wide range in response to complex and often competing hormonal and dietary factors. A combination of transcriptional regulation and rapid protein degradation produces transient elevations of SCD enzyme activity in response to physiologic demands. Dysregulation of SCD has been implicated in non-alcoholic fatty liver disease, hyperlipidemia, and obesity. PMID- 12538077 TI - The role of cytochrome b5 fusion desaturases in the synthesis of polyunsaturated fatty acids. AB - The biosynthetic pathway of polyunsaturated fatty acids (PUFAs) has been the subject of much interest over the last few years. Significant progress has been made in the identification of the enzymes required for PUFA synthesis; in particular, the fatty acid desaturases which are central to this pathway have now all been identified. These "front-end" desaturases are all members of the cytochrome b(5) fusion desaturase superfamily, since they contain an N-terminal domain that is orthologous to the microsomal cytochrome b(5). Examination of the primary sequence relationships between the various PUFA-specific cytochrome b(5) fusion desaturases and related fusion enzymes allows inferences regarding the evolution of this important enzyme class. More importantly, this knowledge helps underpin our understanding of polyunsaturated fatty acid biosynthesis. PMID- 12538078 TI - Essential fatty acid synthesis and its regulation in mammals. AB - The tissue content of highly unsaturated fatty acids (HUFA) such as arachidonic acid and docosahexaenoic acid is maintained in a narrow range by feedback regulation of synthesis. Delta-6 desaturase (D6D) catalyzes the first and rate limiting step of the HUFA synthesis. Recent identification of a human case of D6D deficiency underscores the importance of this pathway. Sterol regulatory element binding protein-1c (SREBP-1c) is a key transcription factor that activates transcription of genes involved with fatty acid synthesis. We recently identified sterol regulatory element (SRE) that is required for activation of the human D6D gene by SREBP-1c. Moreover, the same SRE also mediates the suppression of the D6D gene by HUFA. The identification of SREBP-1c as a key regulator of D6D suggests that the major physiological function of SREBP-1c in liver may be the regulation of phospholipid synthesis rather than triglyceride synthesis. Peroxisome proliferators (PP) induce fatty acid oxidation enzymes and desaturases in rodent liver. However, the induction of desaturases by PP is slower than the induction of oxidation enzymes. This delayed induction may be a compensatory reaction to the increased demand of HUFA caused by increased HUFA oxidation and peroxisome proliferation in PP administration. Recent studies have demonstrated a critical role of peroxisomal beta-oxidation in DHA synthesis, and identified acyl CoA oxidase and D-bifunctional protein as the key enzymes. PMID- 12538079 TI - Hormonal modulation of delta6 and delta5 desaturases: case of diabetes. AB - Animal biosynthesis of high polyunsaturated fatty acids from linoleic, alpha linolenic and oleic acids is mainly modulated by the delta6 and delta5 desaturases through dietary and hormonal stimulated mechanisms. From hormones, only insulin activates both enzymes. In experimental diabetes mellitus type-1, the depressed delta6 desaturase is restored by insulin stimulation of the gene expression of its mRNA. However, cAMP or cycloheximide injection prevents this effect. The depression of delta6 and delta5 desaturases in diabetes is rapidly correlated by lower contents of arachidonic acid and higher contents of linoleic in almost all the tissues except brain. However, docosahexaenoic n-3 acid enhancement, mainly in liver phospholipids, is not explained yet. In experimental non-insulin dependent diabetes, the effect upon the delta6 and delta5 desaturases is not clear. From all other hormones glucagon, adrenaline, glucocorticoids, mineralocorticoids, oestriol, oestradiol, testosterone and ACTH depress both desaturases, and a few hormones: progesterone, cortexolone and pregnanediol are inactive. PMID- 12538080 TI - Steroid hormones and fatty acid desaturases. PMID- 12538081 TI - Recent advances in molecular cloning of fatty acid desaturase genes and the regulation of their expression by dietary vitamin A and retinoic acid. AB - Vitamin A, as an essential micronutrient, is involved in higher animals in embryonic development and postnatal growth, reproduction and maintenance of normal skin, immunity and vision. Recently, studies in vivo and in cell lines have shown that vitamin A and its active metabolite, retinoic acid, regulate the expression of fatty acid desaturases including stearoyl-CoA desaturase and delta 5 desaturase. Whereas the former desaturase catalyzes the formation of monounsaturated from saturated fatty acids, the latter enzyme is involved in the desaturation pathway of dietary essential fatty acids for production of polyunsaturated fatty acids. The reaction products of these desaturases serve as critical regulators in a wide range of physiological processes which include fetal growth and development, reproduction, cell differentiation, immune and inflammatory responses. PMID- 12538082 TI - Biosynthesis of docosahexaenoic acid (DHA, 22:6-4, 7,10,13,16,19): two distinct pathways. AB - Docosahexaenoic acid (DHA) has long been recognized for its beneficial effect in humans, but its biosynthetic pathway has not been clearly established until recently. According to Sprecher, in mammals, DHA is synthesized via a retro conversion process in peroxisomes-the aerobic delta4 desaturation-independent pathway. Recent identification of a Thraustochytrium delta4 desaturase indicates that delta4 desaturation is indeed involved in DHA synthesis in Thraustochytrium. More interestingly, an alternative pathway for DHA biosynthesis-the anaerobic polyketide synthase pathway was also reported recently to occur in Schizochytrium, another member of the Thraustochytriidae. This mini-review attempts to assess the latest research on these distinct pathways for DHA biosynthesis. PMID- 12538083 TI - Regulation of fatty acid desaturation in Bacillus subtilis. AB - The Des pathway of Bacillus subtilis regulates the expression of the acyl-lipid desaturase, Des, thereby controlling the synthesis of unsaturated fatty acids from saturated phospholipid precursors. Activation of this pathway takes place when cells are shifted to low growth temperature or when they are grown in minimal media in the absence of isoleucine supplies. The master switch for the Des pathway is a two-component regulatory system composed of a membrane associated kinase, DesK, and a soluble transcriptional regulator, DesR, which stringently controls transcription of the des gene. We propose that both, a decrease in membrane fluidity at constant temperature and a temperature downshift induce des by the same mechanism, involving the ability of DesK to sense a decrease in membrane fluidity. PMID- 12538084 TI - Effect of H. pylori infection on the expression of cyclooxygenase-2 in human gastric mucosa. AB - Cyclooxygenase-1 is the primary isoform responsible for the production of cytoprotective prostaglandins (PGE(2) and PGI(2)) in the stomach. In contrast COX 2 is induced at the sites of inflammation. Using Helicobacter pylori infection as a model of inflammation, this study was designed to evaluate the effects of H. pylori infection on prostanoid synthesis and expression of COX-2 in human gastric mucosa. Prostaglandin (PGE(2)) and prostacyclin (PGI(2)) synthesis in gastric biopsies obtained from 21 patients undergoing diagnostic endoscopy, were determined. H. pylori was detected by CLO test, histology and culture. Biopsy samples were incubated either with NS-398, selective COX-2 inhibitor or aspirin. Samples were also treated with endotoxin (LPS) in order to induce COX-2 expression. Tissue was also analysed for COX-2 expression in vivo by immunohistochemistry. In 15 out of 21 patients, H. pylori was detected by at least two of the three methods. Higher levels of PGE(2) and PGI(2) were seen in patients infected with H. pylori (191+/-30 and 245+/-88ng/mg protein, respectively) compared with non-infected patients (77+/-17 and 120+/-36ng/mg protein, respectively). There was significant inhibition of PGE(2) and PGI(2) with aspirin in both H. pylori infected (28+/-6.6 and 53+/-43ng/mg, respectively) and in non-infected patients (16+/-7 and 12.5+/-3.5ng/mg protein, respectively). However, NS-398 and LPS did not alter prostaglandin function significantly. Immunohistochemistry in all patients irrespective of Hp status demonstrated expression of COX-2.Lower concentration of constitutive expression of COX-2 was detected in human gastric mucosa by immunohistochemistry, however, H. pylori infection failed to induce COX-2 protein. In addition, increased prostaglandin synthesis in Hp-infected patients appears to be COX-1 mediated rather than COX-2. Furthermore, failure of endotoxaemia-treated sample to produce more PGE(2) in the face of enhanced COX-2 expression in gastric mucosa further suggests that increased prostanoids in human gastric stomach are COX-1 mediated. PMID- 12538085 TI - Association between E-cadherin expression by human colon, bladder and breast cancer cells and the 13-HODE:15-HETE ratio. A possible role of their metastatic potential. AB - The relationship between 15(S)-HETE and 13(S)-HODE from different human tumor cells exposed to n-6 and n-3 essential fatty acids (EFAs) and E-cadherin expression was studied. Colon cancer cells (HRT-18) exposed to gamma linoleic acid (18:3n-6, GLA) and eicosapentaenoic (20:5n-3, EPA) (50microM) showed an increased expression of E-cadherin. Breast cancer (MCF-7) exposed to EPA showed an increment whereas GLA had no effect on E-cadherin expression. No expression of E-cadherin was observed for urothelial cancer (T-24) after GLA or EPA treatment. Significant levels of 15(S)-HETE and 13(S)-HODE were detected after GLA or EPA treatment for all tumor lines. E-cadherin expression was inversely proportional to the 13(S)-HODE:15(S)-HETE ratio when cells were pretreated with GLA or EPA. Nevertheless, the liberation of these metabolites seems to be independent of the E-cadherin expression. The increase in the13(S)-HODE:15(S)-HETE correlates to a decrease in the expression of E-cadherin. Both factors may play a role in metastasis development. PMID- 12538086 TI - COX-2 inhibition potentiates the antiproteinuric effect of enalapril in uninephrectomized SHR. AB - PGE(2) and PGI(2) reduce extracellular matrix deposition and their production is altered after ACE inhibitor (ACEi) treatment. We therefore hypothesized that cyclooxygenase (COX)-2 inhibition would exacerbate renal injury and antagonize the effects of ACEi. To test these hypotheses, WKY and SHR were uninephrectomized (UNX) and treated with either vehicle, enalapril, NS398 or enalapril+NS398. NS398 did not affect systolic blood pressure nor antagonize the antihypertensive effect of enalapril. Urinary protein excretion in UNX WKY was significantly decreased after treatment with either enalapril or NS398. In UNX SHR, enalapril reduced proteinuria, but NS398 alone had no effect. Administration of both drugs, however, further reduced proteinuria. In UNX WKY, treatment with either NS398 alone or both drugs reduced glomerular volume and similar results were observed in SHR. Surprisingly, these results disprove our original hypothesis and suggest that inhibition of COX-2 provides additional renoprotection to that of enalapril alone. PMID- 12538087 TI - Bimatoprost (Lumigan((R))) is an agonist at the cloned human ocular FP prostaglandin receptor: real-time FLIPR-based intracellular Ca(2+) mobilization studies. AB - Bimatoprost is the ethyl amide derivative of 17-phenyl-trinor prostaglandin F(2alpha). Here, we show that bimatoprost (K(i)=9250+/-846nM) and bimatoprost free acid (17-phenyl-trinor prostaglandin F(2alpha); K(i)=59+/-6nM) bind to the FP receptor and displace [(3)H]-travoprost acid, a selective FP agonist. Bimatoprost (EC(50)=3070+/-1330nM), Lumigan((R)) (bimatoprost 0.03% ophthalmic solution; EC(50)=1150+/-93nM) and bimatoprost acid (EC(50)=15+/-3nM) mobilized intracellular Ca(2+) ([Ca(2+)](i)) in <5s in HEK-293 cells expressing the cloned human ciliary body FP receptor on a fluorometric imaging plate reader (FLIPR). Furthermore, agonist effects of bimatoprost and bimatoprost acid were blocked by AL-8810 (11beta-fluoro-15-epi-15-indanyl prostaglandin F(2alpha); K(i)=0.7-2.1 MicroM), an FP receptor-selective antagonist. Therefore, the prodrug bimatoprost and its hydrolytic product, bimatoprost free acid, bind to and activate the human ocular FP prostaglandin receptor to mobilize [Ca(2+)](i), thus behaving as FP receptor agonists. PMID- 12538088 TI - Regional biosynthesis of prostaglandins and hydroxyeicosatetraenoic acids from arachidonic acid in the rat stomach tissue. AB - The present study was conducted to determine regional differences in the biosynthesis of prostaglandins (PGs) and hydroxyeicosatetraenoic acids (HETEs) in the rat stomach tissue (fundus, corpus and pyloric antrum) from radioactive arachidonic acid (AA). The radioactive metabolites were validated by RP-HPLC using non-radioactive AA as substrate. PGE(2) was the major prostanoid in the tissue(.) The relative ratio of PGE(2):PGF(2)alpha:PGD(2) in the whole stomach was 1:0.5:0.1. Regionally, the fundus biosynthesized the largest amount of all three cyclo-oxygenase products. Among the lipoxygenase metabolites, 15S-HETE was the predominant product, while 12S-HETE was found to be the lowest. The relative ratio of 15S-HETE:5S-HETE:12S-HETE in the whole stomach was 1:0.6:0.4. Interestingly, the generation of lipoxygenase products was the highest in the pyloric antrum when compared to fundus or corpus. Thus, the regional differences in the biosyntheses of gastric PGs and monohydroxy fatty acids may be relevant to our understanding of corresponding differences in mucosal resistance or susceptibility to gastric disease. PMID- 12538089 TI - Acyl-CoA thioesterase activity in human placental choriocarcinoma (BeWo), cells: effects of fatty acids. AB - The effects of fatty acids on acyl-CoA thioesterase activity and peroxisome proliferator-activated receptor gamma (PPARgamma), a regulator of lipid metabolism, were investigated in placental choriocarcinoma (BeWo) cells. Substrate preference for acyl-CoA thioesterase was in the following order; gamma linolenoyol-CoA>/=arachidonoyol-CoAz.Gt;palmitoyl-CoA>/=linoleyol-CoA. However, when these cells were incubated with fatty acids, acyl-CoA thioesterase activity was increased by both conjugated linoleic and gamma linolenic acids, but not by docosahexaenoic and eicosapentaenoic acids. In addition, these fatty acids also increased expression of PPARgamma in these cells, suggesting a putative relationship between free fatty acid generated by acyl-CoA thioesterase and expression of PPARgamma. Since expression of PPARgamma is critical for feto placental growth, these fatty acids may be important during pregnancy. PMID- 12538090 TI - Pharmacological evaluation of the novel thromboxane modulator BM-567 (I/II). Effects of BM-567 on platelet function. AB - The aim of this work was to evaluate the effects of BM-567 (N-pentyl-N'-[(2 cyclohexylamino-5-nitrobenzene)sulfonyl]urea), a torasemide derivative, on both thromboxane A(2) (TXA(2)) receptors (TP) and thromboxane synthase of human platelets. The drug affinity for TP receptors of human washed platelets has been determined. In this test, BM-567 showed a high affinity (IC(50): 1.1+/-0.1nM) for the TP receptors in comparison with BM-531 (IC(50): 7.8+/-0.7nM) and sulotroban (IC(50): 931+/-85nM), two TXA(2) antagonists. We also demonstrated that BM-567 prevented platelet aggregation induced by arachidonic acid (AA) (600 microM) (ED(100): 0.20+/-0.10 microM), U-46619, a stable TXA(2) agonist (1 microM) (ED(50): 0.30+/-0.04 microM) and collagen (1microgram ml(-1)) (% of inhibition: 44.3+/-4.3% at 10 microM) and inhibited the second wave of ADP (2microM). Moreover, when BM-567 was incubated in whole blood from healthy donors, the closure time measured by the Platelet Function analyzer (PFA-100((R))) was significantly prolonged (closure time: 215+/-21s) by using collagen/epinephrine cartridges. Finally, at the concentration of 1 microM, BM-567 completely reduced the TXB(2) production from human platelets stimulated with AA (600 microM). These results indicate that BM-567 is a novel combined TXA(2) receptor antagonist and thromboxane synthase inhibitor characterized by a powerful antiplatelet potency. PMID- 12538091 TI - Pharmacological evaluation of the novel thromboxane modulator BM-567 (II/II). Effects of BM-567 on osteogenic sarcoma-cell-induced platelet aggregation. AB - Evidence exists that a large number of tumor cells such as osteosarcoma cells stimulate platelet aggregation, which can be an early step in the metastatic processes of these tumors. Thromboxane A(2) (TXA(2)) is released during platelet aggregation, and it has been suggested that this release may be pathogenic for tumor metastasis for several reasons:Some tumors release large amounts of TXA(2) compared to normal tissue.TXA(2) potentiates tumor growth in culture and increases metastasis in animals.TXA(2) is a potent stimulant of platelet aggregation and causes vascular injuries that may promote implantation of tumor cell-platelet aggregates. If TXA(2) participates in tumor metastasis, it may be hypothesized that TXA(2) inhibitors should decrease tumor metastasis. So, we have evaluated the effects of the original TXA(2) synthase inhibitor and TXA(2) receptor antagonist BM-567 on platelet aggregation induced by osteosarcoma cells using MG-63 tumor cells. Results obtained showed that this drug inhibited both MG 63 tumor-cell-induced platelet aggregation and platelet TXA(2) release following the tumor cell stimulation with IC(50) values of 3.04x10(-7) and 2.51x10(-8)M, respectively. PMID- 12538092 TI - Correlation of the fatty acid composition and fluid property of the cholesteryl esters in the serum of Nigerian children with sickle cell disease and healthy controls. AB - In a previous study conducted in Nigeria, we found that children with sickle cell disease (SCD) had exceedingly low total serum cholesterol levels (mean=100 102mg/dl). The fact that significant reductions in the levels of certain polyunsaturated fatty acids (PUFA) have been documented in the serum phospholipids of these same SCD subjects led us to inquire as to the fatty acid composition of the cholesteryl esters (CE) in their serum. Lecithin:cholesterol acyl transferase (LCAT), the enzyme in blood that catalyzes the reaction in which tissue cholesterol is acylated prior to its removal from cell membranes, is relatively specific for certain PUFA. CE in blood serum from 43 male and 42 female children with SCD, ages 4-18 years, and equal numbers of age- and gender matched controls were analyzed for their fatty acid composition. Relative to the non-SCD controls, the CE of the SCD subjects contained 9% less linoleic acid, 16% less arachidonic acid, 40% less alpha-linolenic acid, 50% less eicosapentaenoic acid, and 36% less docosahexaenoic acid, but 15% more palmitic acid and 10% more oleic acid. Overall, the acyl chains of the CE of the SCD subjects were less fluid than those of the controls, as determined by comparison of their mean melting points (MMP) and double bond indices (DBI). MMP and DBI were both estimated from the individual constituent fatty acids comprising the CE acyl chains. The strongest correlations between MMP and fatty acid mole percent were seen with palmitic acid and linoleic acid. These results show that the fatty acid composition of the serum CE of children with SCD is abnormal relative to controls who do not have this hematologic disorder. We speculate that suboptimal fatty acid nutrition in Nigerian children with SCD compromises their ability to remove cholesterol from their tissues due to preference of the LCAT enzyme for PUFA, thereby accounting, in part at least, for the low total serum cholesterol levels one finds in children with SCD. PMID- 12538093 TI - [Should fundoplication be added to Heller's myotomy?]. PMID- 12538094 TI - [Portal cavenorma: diagnosis, aetiologies and consequences]. AB - Portal cavernoma is a network of veins whose caliber, initially millimetric or microscopic, is increased and which contain hepatopedal portal blood. It results from occlusion, thrombotic and always chronic, of the extra-hepatic portal system. Diagnosis is mainly done by imaging. Clinical signs of portal cavernoma are usually related to extra-hepatic portal hypertension (hematemesis due to rupture of oeso-gastric varices, splenomegaly, rectal bleeding from ano-rectal varices, growth retardation in children) and sometimes to the cause of portal hypertension (abdominal pain, venous bowel infarction). Occurrence of portal thrombosis is often the conjunction of a local cause and a prothrombotic disorder which must be systematically detected. Biliary consequences of cavernoma are related to compression of common bile duct and are usually asymptomatic. In case of jaundice or cholangitis, portal decompression by portosystemic shunt can be performed to treat both biliary symptoms and portal hypertension. PMID- 12538095 TI - [Targeted parathyroid surgery: a series of 100 consecutive patients]. AB - INTRODUCTION: The treatment of primary hyperparathyroidism has long required a bilateral neck exploration to localize all parathyroid glands. New techniques in pre-operative imaging (technetium-99m-sestamibi scanning, high resolution ultrasonography) have allowed an accurate localization of pathological glands. Moreover, the intraoperative intact parathyroid hormone assay (iPTH) now permits to confirm the resection of all hyper-secreting glands. MATERIAL AND METHODS: Two hundred and twenty patients underwent parathyroid operations during the last 3 years, by different minimally invasive techniques. Pre- and intra-operative data, as well as the follow-up were recorded prospectively. The results of the last 100 unilateral approaches, performed for primary hyperparathyroidism, were analyzed. RESULTS: There were 78 women and 22 men, with a mean age of 57 +/- 15 years (25 to 92 years). Mean operative time was 44 +/- 25 min. Ninety-three solitary adenomas, three double adenomas, three carcinomas and one hyperplasia were resected. Two patients required a bilateral exploration. There was one intra operative complication (pneumothorax) during the resection of a mediastinal gland. Three post-operative complications occurred, with 2 hematomas and one transient recurrent laryngeal nerve paralysis. Median hospital stay was 2 +/- 7,5 hours (1-72 hours). All patients were cured following the operation, as predicted by the intra-operative iPTH assays. CONCLUSION: These results suggest that unilateral, targeted, neck explorations offer a cure rate similar to those obtained by bilateral neck explorations. Moreover, this operation can be performed under local anesthesia, with a short hospital stay and recovery time. The morbidity rate is low but should be addressed by large-scale comparative studies. PMID- 12538096 TI - [Oesophageal cancer in patients with head and neck cancers: therapeutic implications]. AB - AIM OF THE STUDY: To determine therapeutic and prognostic implications of an associated head and neck primary cancer in patients undergoing oesophagectomy for squamous cell carcinoma of the oesophagus. PATIENTS AND METHODS: Between 1982 and 2000, 868 patients with oesophageal cancer were operated in our institution, including 78 (9%) who underwent oesophagectomy for associated oesophageal and head and neck cancers; the latter was synchronous (n = 52) or anterior metachronous (n = 26). Influence of head and neck cancer on the treatment of oesophageal carcinoma was analysed retrospectively in terms of surgical therapeutic strategy and survival. RESULTS: Oesophageal resection consisted of oeso-pharyngolaryngectomy (n = 14, 17.9%), subtotal oesophagectomy (n = 62, 79.5%) and cervical oesophagectomy (n = 2, 2.6%). Radical resection (R0) was obtained in 85% of cases. Postoperative mortality rate was 5 % (4/78). Main complications were pulmonary (18% = 14/78) and anastomotic leaks (14% = 11/78), all of them cervical. Follow-up (mean = 25 +/- 27 months) was complete for all 78 patients. Five-year survival after R0 resection was 25%. Survival pronostic factors were denutrition, complete resection, and pT status of oesophageal tumor. CONCLUSION: In patients with associated carcinomas of oesophagus and head and neck, agressive treatment -including an oesophagectomy- allowed a 5-year survival rate more than 25% without increased mortality or morbidity rates, compared with patients operated on for isolated oesophageal carcinoma. PMID- 12538097 TI - [Post-operative small bowel obstruction: spiral computed tomography]. AB - INTRODUCTION: The aim of this study was to evaluate prospectively the impact of the routine use of abdominal spiral computed tomography (SCT) in patients with postoperative small bowel obstruction (SBO) for whom initial conservative treatment was proposed. PATIENTS AND METHODS: We have compared the management of SBO in patients with clinical stable condition in two successive periods : from 1989 to 1998, 127 patients (preSCT group) for whom management was based on standard clinical-biological-radiological assessment (CBRA) et from 1999 to 2000, 30 patients (SCT group) for whom management included SCT. The decision of surgical team was correlated with the type of small bowel obstruction at laparotomy : closed-loop obstruction without intestinal necrosis (true-positive), intestinal necrosis as a consequence of delayed diagnosis defined as false negative, diffuse adhesion defined as false-positive et patient non operated defined as true-negative. RESULTS: Among the 127 patients from the preSCT group, 87 were treated conservatively and 40 were operated : SBO with closed-loop obstruction without intestinal necrosis (n = 29,72%), SBO with diffuse adhesion (n = 4, 10%) and SBO with intestinal necrosis (n = 7, 17%). Among the 30 patients from the SCT group, 16 were treated conservatively and 14 were operated: SBO with closed-loop obstruction without intestinal necrosis (n = 8, 57%), SBO with diffuse adhesion (n = 6,43%) and SBO with intestinal necrosis (n = 0,0%; NS). Both groups were similar for rates of patients with SBO with or without necrosis and rate of patients treated conservatively (NS). In SCT group, there was significantly more patients operated for diffuse adhesions (p < 0,01). Negative predictive value of CBRA + TDM was significantly higher than those of CBRA alone (p = 0,041). CONCLUSION: Due to a very high sensibility, TDM increase probably the rate of early laparotomies, maybe unnecessary, in patients without any sign of SBO due to closed-loop obstruction. Thus, systematic use of TDM in patients with clinical suspicion of SBO remains to be evaluated. PMID- 12538098 TI - [Heller's myotomy without fundoplication: a series of 123 patients]. AB - INTRODUCTION: Surgical treatment of achalasia of lower oesophageal sphincter is Heller's myotomy, usually associated with a fundoplication due to an high risk of postoperative gastro-oesophageal reflux. The value of this fundoplication is discussed. The aim of this study was to evaluate retrospectively the results of Heller's myotomy without fundoplication but performed according to a precise technique preventing postoperative reflux. PATIENTS AND METHODS: Between 1975 and 1999, 123 patients underwent Heller's myotomy without systematic fundoplication. Diagnosis of achalasia was performed clinically and confirmed by investigations: baryum meal, fibroscopy and manometry. Myotomy was performed through an abdominal approach in 117 (95%) patients. Dissection preserved fixity of abdominal oesophagus in all cases, particularly its posterior meso. Myotomy was performed on abdominal oesophagus but not below the cardia. Posterior fundoplication was associated in 2 patients. RESULTS: One patient (0,8%) died from massive aspiration. Morbidity (1,6%) consisted in one peritonitis and one postoperative occlusion. At follow-up (mean = 5 years; range: 1-20), functional results were satisfying (excellent and good) in 112 (92%) patients. Seven patients (6%) developed postoperative reflux, including one who need surgical treatment. Dysphagia persisted in 3 patients (2%) who had to be reoperated. CONCLUSION: Results of this series show that systematic fundoplication is not necessary in Heller's myotomy for achalasia of lower oesophageal sphincter. PMID- 12538099 TI - [Compression of the inferior vena cava with thrombus: a rare complication of solitary liver cyst]. AB - Because of the increase of ultrasonographic exams, nonparasitic cyst of the liver is frequently encountered. Some giant cysts have complications. The surgical treatment for complicated forms consists in resection of the protruding part. We report a case of a giant non parasitic cyst of the liver which was responsible of a compression of the inferior vena caval with thrombus. The resection of the thrombus was performed after a double control of the vena cava and a control of the aorta. The fenestration of the cyst was performed in a second time. PMID- 12538100 TI - [Acute clinical pancreatitis following selective transcatheter arterial chemoembolization of hepatocellular carcinoma]. AB - Acute pancreatitis can complicate non-selective transcatheter arterial embolization of hepatocellular carcinoma with an incidence ranging from 1,7% (acute clinical pancreatitis) to 40% (biological pancreatitis). This complication is thought to be related to embolization of extrahepatic arterial collaterals. We report herein a case of acute clinical pancreatitis developing within 24 hours after a second course of selective transcatheter arterial chemo-embolization into the proper hepatic artery. Neither anatomical arterial variation nor particular risk factor for acute pancreatitis could be identified. This complication is unusual after selective arterial embolization. Because it may clinically mimick a postembolization syndrome, dosage of serum pancreatic enzymes should be performed systematically in case of abdominal pain following chemoembolization. PMID- 12538101 TI - [The surgical treatment of liver metastasis of carcinoid tumors]. AB - Liver metastasis of carcinoid tumors from the digestive tract are rare entities, with characteristic symptoms and slow evolution. Symptomatic metastases may justify surgical resection. Herein, we report a series of nine patients, who underwent 14 hepatic resections (6 major and 8 minor), which was radically operated in 71%. The postoperative mortality was zero and the morbidity was 7%. Mean follow-up was 82 month (median 75 months) and global survival rate was 89%. Two patients are in complete remission. Our experience suggest that surgical hepatic resection, when feasible, is efficient and associated with prolonged survival despite a high recurrence rate. PMID- 12538102 TI - [Interest of a programme of monitoring of infections' incidence of the operative site in digestive surgery]. PMID- 12538103 TI - [The suprahepatic approach. An easy laparoscopic access to the hiatus in obese patients]. AB - The conventional laparoscopic access to the gastroesophageal junction involves the retraction the left hepatic lobe upward. This approach could be particularly difficult in obese patients because of a huge fatty liver. The technique described here aims to overcome this difficulty be using a suprahepatic route by displacing the left lobe downward after dividing the left triangular ligament. This approach allows a good exposure of the gastroesophageal junction to perform a fundoplication or a gastric banding. PMID- 12538104 TI - [Compartment syndrome after proctectomy]. PMID- 12538110 TI - [Digest of the ANZJ Surg 2001]. PMID- 12538111 TI - Six-month angiographic evaluation of beating-heart coronary arterial graft interrupted anastomoses using the coalescent U-CLIP anastomotic device: a prospective clinical study. AB - BACKGROUND: Interrupted suture technique avoids the "purse string" and puckering effects frequently seen with continuous suture techniques and should represent the standard of care in the creation of high-quality vascular anastomoses. This clinical study evaluated the safety and effectiveness of a self-closing surgical clip (Coalescent Surgical U-CLIP Anastomotic Device [U-CLIP]) designed to facilitate this interrupted technique. Left internal mammary artery (LIMA) to left anterior descending (LAD) coronary bypass grafting was studied. METHODS: Eighteen patients meeting inclusion criteria were enrolled (October 2000 through September 2001) into this prospective study. Anastomoses were performed using a beating-heart median sternotomy procedure (off-pump coronary artery bypass) in 17 cases (94%) and a minimally invasive beating-heart procedure (minimally invasive direct coronary artery bypass [MIDCAB]) in one case (6%). Six-month follow-up was completed on 18 patients (100%), with angiograms performed on 17 patients (94%) at a mean of 179 days (range, 168-191 days). Qualitative and quantitative angiographic assessment was performed by an independent core laboratory. RESULTS: The U-CLIP was used for 18 LIMA-to-LAD interrupted anastomoses without the requirement for knot tying or suture management and with no device-related morbidity or mortality. Mean LIMA-to-LAD anastomosis time was 8.6 minutes (range, 5-14 minutes). All anastomoses were FitzGibbon grade A at 6 months postprocedure. Quantitative analysis showed mean luminal diameters proximal to the anastomosis of 2.32 mm, at the anastomosis of 2.25 mm, and immediately distal to anastomosis of 1.99 mm. The average ratio of anastomosis to LAD diameter was 1.17 (range, 0.93- 1.93). Anastomotic stenosis as a percentage of average LIMA/LAD diameter was a negative 4.2%, comparing favorably with the 23% to 24% reported in the POEM (Patency, Outcomes, Economics of MIDCAB) study. CONCLUSIONS: The interrupted technique, facilitated by a self-closing anastomotic clip, yielded 6-month follow up and angiographic results that compared favorably with results of other published studies. PMID- 12538112 TI - Clinical investigation: endoscopic coronary artery bypass grafting with robotic assistance. AB - BACKGROUND: The current study reviews clinical feasibility experiences evaluating safety and efficacy of using robotic assistance to create a left internal mammary artery to left anterior descending artery (LIMA-to-LAD) anastomosis. METHODS: Between August and November 1999, 9 patients (aged 54-73 years) underwent robotically assisted endoscopic coronary artery bypass grafting (E-CABG) after institutional review board approval and informed consent were obtained. The robotics were transthoracically introduced in the fifth and sixth intercostal spaces. The LIMA-to-LAD anastomoses were endoscopically constructed with robotic assistance, and patency was assessed by flow measurement. RESULTS: Each anastomosis was performed robotically, without necessity for intraoperative intervention with traditional techniques. Robotic anastomosis times averaged 29.05 minutes. The robotic system added on average 41.28 minutes to the procedure. LIMA flows prior to anastomoses measured from 11.2 to 29.2 mL/min. LIMA flow measurements following anastomoses averaged 42.07 mL/min. There were no deaths or perioperative myocardial infarctions. CONCLUSIONS: Although E-CABG is an exhaustive and technically demanding procedure, it is feasible for a computerenhanced robotic telemanipulation system to safely and effectively provide substantial assistance to the surgeon completing a thoracoscopic coronary anastomosis. PMID- 12538113 TI - Angiosarcoma of the right ventricle: a rare encounter. AB - We report a case of angiosarcoma involving the right ventricle. The patient was seen in our Cardiology Department and subsequently referred to our unit for surgery. He gave a 1-week history of lethargy, chest pain, breathlessness on exertion, fevers, and night sweats. Echocardiography and computed tomography of the chest showed a large pericardial effusion with multiple densities, raising suspicions of a hemorrhagic effusion. Surgical exploration showed an epicardial mass. Histopathology revealed angiosarcoma. PMID- 12538114 TI - A video-assisted thoracoscopic technique to encircle the four pulmonary veins: a new surgical intervention for atrial fibrillation ablation. AB - BACKGROUND: Atrial fibrillation (AF) is the most common of the so-called benign arrhythmias. It affects not only life expectancy but also quality of life. Until recently, surgeons have most often encountered AF in association with ischemic or valvular disease but rarely as lone atrial fibrillation (LAF). For the subset of LAF patients, a minimally invasive procedure is recommended. METHODS: Using an animal model, we have developed a video-assisted thoracoscopic approach to atrial ablation whereby the ablation is performed encircling the four pulmonary veins as through a median sternotomy. RESULTS: Fifteen animals were used, and in 5 a complete encircling of the pulmonary veins was accomplished using the thoracoscopic approach. DISCUSSION: Video-assisted thoracoscopy is a feasible and safe approach for epicardial pulmonary vein ablation. This technique offers the option of surgery to a class of patients who are resistant to medical therapy but for whom the presence of LAF contraindicates the open chest approach. PMID- 12538115 TI - Myoblasts survive intracardiac transfer and divide further after transplantation. AB - OBJECTIVE: Skeletal myoblasts have been proven to survive transplantation into myocardial scar tissue. The objective of this study was to evaluate whether these cells can also be transferred into vital myocardium and maintain their ability for cell division after transplantation. In addition, an intravital fluorescence dye for marking these cells was evaluated. MATERIAL AND METHODS: Skeletal myoblasts were harvested from male Lewis rats (n = 6) and then expanded in culture. Before implantation, these cells were trypsinized and labeled using an intravital fluorescence dye (PKH-26). Syngenic myoblast transfer to recipient female Lewis rats (n = 36) was used to simulate autologous transplantation. Under general anesthesia, the rats received injections of 106 myoblasts via a subxyphoidal approach into the apex of the heart. The animals were then divided into 3 groups (n = 10 each). The animals were sacrificed at several time points, and the hearts were harvested for histologic examination: group A, 7 days postoperatively; group B, 14 days postoperatively; and group C, 28 days postoperatively. An additional group, group D (n = 6), served as a control group; these animals were injected with only cell medium. Corresponding to the study groups, 2 animals of this control group were sacrificed at each time point, and the hearts were explanted. At histological examination, 8- m sections were investigated to identify surviving stained cells. For further evaluation, the sections were stained using monoclonal antibodies against n-cam, desmin, and a- actin. RESULTS: No fluorescing cells were found in any hearts of rats in the control group. Surviving fluorescing myoblasts were found in 9 of 10 hearts of groups A and C and 8 of 10 hearts of group B. Labeled myoblasts were located in the intercellular spaces between the myocardial fibers. Fibrotic or inflammatory reactions could not be identified around the injection site in any hearts of the study groups. Immunohistochemical staining results showed that the labeled cells expressed n-cam, desmin, and a-actin. The myoblasts had regained their physiologic structures and had started to form myofibers. In groups B and C, more n-cam-positive cells than labeled cells were found, indicating further cell division. CONCLUSIONS: Intravital fluorescence staining with PKH- 26 dye proved to be an easy and reliable method for identifying cells after cellular transplantation. Myoblasts survived an intracardiac transfer, regaining their physiologic structures and maintaining their ability for further cell division. PMID- 12538116 TI - Clinical experience with the CorLink device for proximal anastomosis of the saphenous vein to the aorta: a clinical, prospective, and randomized study. AB - BACKGROUND: Avoiding tangential clamping of the ascending aorta during coronary bypass operations reduces the trauma to the aorta and may avoid local particulate embolization. METHODS: From December 2000 to May 2001, 21 male patients, mean age 64.1 +/- 7.2 years (range, 46-76 years), with coronary artery 2-vessel (n = 3) and 3-vessel (n = 18) disease were divided randomly into 2 groups and underwent myocardial revascularization. In 11 patients an aorta-saphenous vein graft anastomosis was performed with the CorLink device for anastomosis between the saphenous vein and the ascending aorta. Ten patients served as control subjects. In these patients the central bypass anastomosis was performed with a 6-0 running suture. Clinical follow-up was performed 1 month and 3 months postoperatively. Six months after surgery, multislice computed tomography was performed to evaluate bypass patency for all patients. RESULTS: Mean number of study vessels was 1.2 +/- 0.4 in the CorLink group and 1.5 +/- 0.5 in the control group. In the CorLink group, 13 additional arterial and vein grafts were performed, and in the suture control group 15 additional mammary artery grafts were carried out. No intraoperative complications occurred. In 2 CorLink anastomoses an additional stitch was necessary because of minor bleeding. Follow-up was carried out at 6 months with multislice computed tomography for all patients and showed only 1 study vessel occlusion in the CorLink group. All 62 other bypass grafts were revealed to be patent and had anastomoses of good quality. CONCLUSION: Our experience suggests that the CorLink device is a safe and effective technique for anastomosis between saphenous vein grafts and the ascending aorta. The CorLink device could be used for totally endoscopic coronary bypass operations. Further randomized studies enrolling a larger number of patients are necessary to determine which patients may benefit the most from this procedure. PMID- 12538117 TI - Cardiac surgery in patients with heparin-induced thrombocytopenia using preoperatively determined dosages of iloprost. AB - BACKGROUND: Patients with preoperatively diagnosed type II heparin-induced thrombocytopenia (HIT) scheduled for cardiopulmonary bypass (CPB) present a challenge in their intraoperative anticoagulation management because re-exposure to heparin may result in profound thrombocytopenia, intravascular thromboses, bleeding, and even death. Iloprost, a prostacyclin analogue that reversibly inhibits platelet aggregation, has been suggested as a management approach in such cases. The purpose of this study was to assess and confirm the efficacy of a perioperative intravenous iloprost infusion in preventing thromboembolic complications in patients with type II HIT undergoing cardiac surgery and requiring the use of heparin and CPB. METHODS: During a one-and-a-half-year period, 22 patients with type II HIT presented at the Cardiac Surgery Service of the Onassis Cardiac Center in Athens. In these patients, platelet aggregation test results were found strongly positive at heparin serum concentrations corresponding to those achieved during CPB. Iloprost was used in a preoperatively, in vitro-determined, patient-specific concentration that was assessed and modified perioperatively depending on its in vivo effect on platelet aggregation as opposed to the conventional constant rate. RESULTS: In the 22 patients, the preoperatively determined concentration of iloprost seemed to correlate well with the in vivo interruption of platelet aggregation, as tested by a perioperative heparin-induced platelet aggregation (HIPA) assay, and in only 3 cases (14%) was the rate of iloprost infusion increased. The patients' platelet counts, which were evaluated peri- and postoperatively, were preserved with no statistically significant fluctuations. Postoperative bleeding was within normal limits and no thrombotic episodes or other complications were reported. CONCLUSION: Although a number of alternative anticoagulation methods, such as the use of another anticoagulant (danaparoid sodium and recombinant hirudin) or the preoperative use of a defibrinogenating agent (ancorod), have been suggested for patients with type II HIT requiring anticoagulation during CPB, the use of heparin associated with a potent platelet inhibitor such as the prostacyclin analog iloprost is, as this study confirmed, the only to-date safe and effective choice. PMID- 12538118 TI - Midterm angiographic results of off-pump coronary artery bypass grafting. AB - BACKGROUND: As off-pump coronary artery bypass grafting (OPCAB) has gained wide attention, the quality of surgical revascularization, which showed favorable initial results, is being frequently questioned. This study was undertaken to assess the midterm outcome of beating-heart coronary anastomosis. METHODS: Seventy-four of 315 patients who underwent beating-heart coronary artery bypass grafting via median sternotomy between March 1994 and December 1995 were randomly selected for angiographic assessment. RESULTS: The mean period of control angiography was 50.1 +/- 22.6 months (range, 22 to 83 months). A total of 109 (1.52 +/- 0.55) anastomosis procedures were performed in 74 patients; 38 had single-vessel disease, and 36 had double-vessel disease or disease involving more than 2 vessels. There were no perioperative returns to cardiopulmonary bypass, no major complications, and no hospital deaths. The causes of the need for occlusion included 2 (2.5%) instances of left internal mammary artery-left anterior descending artery anastomosis; 2 (2.5%) of anastomotic site stenosis in left internal mammary artery-left anterior descending artery anastomosis; 7 (19.4%) of saphenous vein graft anastomosis; and 3 (8.3%) of anastomotic site stenosis of saphenous vein graft. Statistical analysis revealed hypercholesterolemia as an independent predictor for graft occlusion (P =.014). The patency rates were not affected by endarterectomy, length of the anastomosed segment, or coronary artery structure. Event-free survival was 73.61% and myocardial infarction-free survival was 91.67%. Reintervention and reoperation rates were 24.3% (18 instances) and 1.4% (1 instance), respectively. CONCLUSIONS: Our results were encouraging for OPCAB, supporting its safety and effectiveness, patency rates and clinical outcome comparable to those of cardiopulmonary bypass, and overall benefits such as reduced hospital costs and postoperative length of stay. PMID- 12538119 TI - Near-infrared fluorescence coronary angiography: a new noninvasive technology for intraoperative graft patency control. AB - BACKGROUND: Intraoperative graft patency verification is of major clinical importance for quality control after coronary artery bypass grafting (CABG), especially if surgery is performed on the beating heart. This is one of the first reports of fluorescence coronary angiography (FCA) using the dye indocyanine green (ICG), a noninvasive technology for direct visualization of coronary arteries, bypass grafts, and myocardial perfusion. METHODS: Twenty-three domestic pigs (weight, 45-72 kg) underwent FCA of the left anterior descending coronary artery (LAD). In the first group (n = 6 pigs), FCA was used to visualize the native coronary vessels and myocardial perfusion. In the second group (n = 8 pigs), 14 stenoses of various degrees and 4 total vessel occlusions were created by snares on different segments of the LAD, and FCA was used to visualize the effects of these obstructions. In the third group (n = 9 pigs), a coronary bypass procedure on the beating heart was performed by a left internal mammary artery or a human saphenous vein graft to the LAD, and FCA was used to visualize graft patency. Three pigs were removed from the study because of ventricular fibrillation. ICG was intravenously applied, and the heart was illuminated with near-infrared light emitted by laser diodes. The fluorescence emission was detected by an adapted charge-coupled device camera system. The images were displayed in real time on a high-resolution monitor. Subsequently, images obtained with FCA were compared to those obtained with coronary angiography (n = 10 pigs). RESULTS: In all cases, high-quality FCA images of coronary arteries and myocardial perfusion were obtained. All stenoses resulted in an impairment of the myocardial perfusion visualized by FCA. Occlusion of the LAD or the diagonal branch resulted in a total perfusion defect of the corresponding anterior myocardial wall with immediate reperfusion after releasing the snare. In 5 cases a patent bypass graft with an apparent homogenous perfusion of the corresponding myocardium was detectable. In one procedure, FCA images indicated total occlusion of the bypass graft and a total perfusion deficit in the distal LAD region. Correlation between FCA and coronary angiography in detection of stenoses and graft patency was excellent. CONCLUSION: With the fluorescence technique using ICG, visualization of blood flow in coronary vessels and bypass grafts, as well as of myocardial perfusion, is feasible. FCA is a highly sensitive and reproducible method and an excellent technique for intraoperative quality control in CABG. PMID- 12538120 TI - Direct aortic cannulation in minimally invasive mitral-valve operations. AB - The minimally invasive Port-Access (Heartport, Redwood City, CA) approach in mitral-valve operations originally required femoral arterial cannulation, which is considered a disadvantage, especially in patients with peripheral vessel disease. In this study 20 patients were prospectively randomized into 2 groups, to undergo either standard femoral (group A) or direct aortic cannulation (group B). Pre- and postoperative data as well as markers for myocardial damage were assessed. Postoperatively, patients of group B showed lower levels of damage, indicating that direct aortic cannulation might provide better myocardial protection. Furthermore, the direct aortic cannulation technique may eliminate complications associated with the standard femoral artery cannulation. PMID- 12538121 TI - A new thermoregulation system for maintaining perioperative normothermia and attenuating myocardial injury in off-pump coronary artery bypass surgery. AB - BACKGROUND: Most patients undergoing coronary artery bypass surgery demonstrate perioperative mild-to-moderate hypothermia (<36 degrees C). Patients undergoing off-pump coronary artery bypass (OPCAB) grafting may become even more severely hypothermic for want of cardiopulmonary bypass rewarming. One consequence is increased circulating catecholamine levels that induce an elevated systemic vascular resistance (SVR), which causes a subsequent deterioration in cardiac output. MATERIALS AND METHODS: We assessed the ability of the Allon thermoregulatory (AT) system to maintain normothermia and its impact on hemodynamics and myocardial function in patients undergoing OPCAB surgery. In this study, the first 60 of 120 suitable patients were assigned to AT (n = 40) or routine thermal care (RTC) (n = 20). Core body temperature, cardiac index (CI), SVR, and cardiac-specific troponin I (cTnI) were analyzed perioperatively for patients in both groups. RESULTS: Core body temperature was significantly higher in the AT group (from 36.1 degrees C +/- 0.5 degrees C at induction of anesthesia to 37 degrees C +/- 0.5 degrees C during surgery) than in the RTC group (from 35.8 degrees C +/- 0.4 degrees C to 35.2 degrees C +/- 0.8 degrees C, respectively; P <.01). SVR was significantly lower, and CI was greater (at comparable time points), whereas cTnI levels in the AT group were lower than in the RTC group from the end of surgery until 24 hours postoperatively (7.4 +/- 17.7 g/L versus 31.9 +/- 47.4 g/L; P =.03). These findings indicate the possibility for less ischemic damage sustained intraoperatively in the AT group. CONCLUSIONS: Maintenance of perioperative normothermia (36.5 degrees C-37.5 degrees C) during OPCAB procedures can be efficiently achieved with the Allon thermoregulation system. The system was found to be superior to other routinely used methods of temperature maintenance. Benefits may include lowering afterload (as expressed by reduced SVR), an improved CI, and attenuation of myocardial injury (as assessed by cTnI levels). PMID- 12538122 TI - Accelerated recovery after cardiac operations. AB - BACKGROUND: The accelerated-recovery approach, involving early extubation, early mobility, decreased duration of intensive care unit stay, and decreased duration of hospitalization has recently become a controversial issue in cardiac surgery. METHODS: We investigated timing of extubation, length of intensive care unit stay, and duration of hospitalization in 225 consecutive cardiac surgery patients. Of the 225 patients, 139 were male and 86 were female; average age was 49.73 +/- 16.95 years. Coronary artery bypass grafting was performed in 127 patients; 65 patients underwent aortic and/or mitral or pulmonary valvular operations; 5 patients underwent valvular plus coronary artery operations; and in 28 patients surgical interventions for congenital anomalies were carried out. RESULTS: The accelerated-recovery approach could be applied in 169 of the 225 cases (75.11%). Accelerated-recovery patients were extubated after an average of 3.97 +/- 1.59 hours, and the average duration of stay in the intensive care unit was 20.93 +/- 2.44 hours for these patients. Patients were discharged if they met all of the following criteria: hemodynamic stability, cooperativeness, ability to initiate walking exercises within wards, lack of pathology in laboratory investigations, and psychological readiness for discharge. Mean duration of hospitalization for accelerated-recovery patients was 4.24 +/- 0.75 days. Two patients (1.18%) who were extubated within the first 6 hours required reintubation. Four patients (2.36%) who were sent to the wards returned to intensive care unit due to various reasons and 6 (3.55%) of the discharged patients were rehospitalized. CONCLUSIONS: Approaches for decreasing duration of intubation, intensive care unit stay and hospitalization may be applied in elective and uncomplicated cardiac surgical interventions with short duration of aortic cross-clamping and cardiopulmonary bypass, without risking patients. Frequencies of reintubation, return to intensive care unit, and rehospitalization are quite low with this approach. PMID- 12538123 TI - Effects of seasonal variations on coronary artery surgery. AB - BACKGROUND: In this study, we compared profiles and early results of patients who underwent coronary artery bypass graft surgery (CABG) during the winter season with those who underwent CABG during the summer season. We also investigated whether possible seasonal variations in performance of health care professionals had any effects on surgical outcome. METHODS: The data from patients who had undergone CABG surgery in the winter (group A) and summer (group B) seasons of the period from December 1998 to August 2001 were analyzed retrospectively. Preoperative, perioperative, and postoperative data were compared. Preoperative factors analyzed included sex, age, diabetes mellitus, hypertension, New York Heart Association status, unstable angina pectoris, and left ventricle ejection fraction. Perioperative factors included graft number, internal mammary artery use, cross-clamp time, and cardiopulmonary bypass time. Postoperative factors included mediastinal reexploration, blood use, inotropic support, intraaortic balloon pump support, perioperative myocardial infarctus, cardiac arrest, infection and other complications, duration of hospital stay, and early mortality. RESULTS: The preoperative demographic data were identical in the 2 groups, with the exception of incidence rate of hypertension (26.8% in group A versus 15.7% in group B, P <.01). The incidence rates were higher in group B than group A for postoperative infection (8.8% versus 5.2%, P <.05), mediastinal reexploration for bleeding (6.9% versus 4.2%, P <.05), and transfusion blood use (7.3 +/- 6.2 U/patient versus 6.0 +/- 3.9 U/patient, P <.05). There were no differences between the 2 groups in early mortality rates. CONCLUSION: Despite the fact that frequency and occurrence of cardiovascular events traditionally have been reported to be higher in the winter than the summer, our data show no major differences in early surgical outcome among those patients who had undergone CABG in the winter or summer. We did not encounter any seasonal patterns. However, an interesting finding was that the patients who underwent surgery in the summer had a higher incidence of infection and bleeding. PMID- 12538124 TI - Is hybrid coronary revascularization favored by cardiologists or cardiac surgeons? AB - We investigated the present use of integrated coronary revascularization (ICR) by interviewing a sample of United States invasive cardiologists and cardiac surgeons. Both groups still favor left internal mammary artery (LIMA) grafting to revascularize the left anterior descending (LAD) coronary artery. There remains a lack of exposure to and acceptance of ICR, especially for surgeons. We report the findings of this national survey of 180 cardiologists and 160 surgeons, as they may serve as an indicator of the current opinions about ICR and its future applicability as a standard method of coronary artery revascularization. We discuss the limited popularity of minimally invasive hybrid procedures and the importance of further exposing cardiologists and surgeons to ICR. PMID- 12538125 TI - Graft repair of a pulmonary artery aneurysm. AB - Aneurysm of the main pulmonary artery is a rare anomaly with a poorly understood pathogenesis. We report the successful surgical resection and repair of a pulmonary artery aneurysm using aneurysmectomy and replacement of the pulmonary artery with Dacron, a procedure with excellent longterm outcome that eliminates the risk of recurrence. The 65-year-old male patient presented with a marked decrease in exercise tolerance and worsening dyspnea. He had a history of pulmonary stenosis diagnosed during childhood, episodic shortness of breath beginning during his teenage years, hypertension, and smoking. On physical examination, the patient appeared generally well. Computed tomographic scan of the chest revealed an 8-cm aneurysm involving the main pulmonary artery and pulmonary artery bifurcation. Coronary angiography showed a massive aneurysm of the main and left pulmonary arteries, with mild dilation of the proximal right pulmonary artery. Surgery was performed through a median sternotomy with the use of normothermic, bicaval cardiopulmonary bypass. The aneurysm of the main pulmonary artery and dilated bifurcation of the pulmonary artery was excised and reconstruction was performed using a 22-mm Dacron graft. The patient's postoperative course was uneventful. He was discharged to home on postoperative day 5 and was well at 1-month follow-up. PMID- 12538126 TI - What will be the impact of drug-eluting stents on hybrid coronary revascularization? PMID- 12538127 TI - Cell transplantation--a potential therapy for cardiac repair in the future? AB - PURPOSE: Adult human myocardium cannot regenerate because cardiac muscle cells do not reenter the cell cycle. Myoblasts, cardiomyocytes, and stem cell-derived cardiomyocytes have been transplanted in experimental settings to replace lost myocardial tissue. The purpose of this paper is to review the experimental data about cell transfer onto myocardium and to highlight the advantages of the particular cell types used. BACKGROUND AND METHODS: Myoblasts or satellite cells are precursor cells attached to skeletal muscle fibers. The transfer of these cells onto damaged myocardium was demonstrated successfully in several animal models. Because myoblasts can be expanded in culture, a large number of cells can be obtained from only a small skeletal muscle specimen. These cells could be delivered locally by injection within a damaged myocardial area or by coronary infusion. However, only one group has been able to show an improvement in myocardial function after myoblast transfer. The second type of cells used experimentally for cell transfer were fetal cardiomyocytes. Fetal cardiomyocytes retain the ability to divide and therefore can be expanded in culture. The cells were integrated into the myocardial tissue, differentiated into normal cardiac muscle cells, and formed intercellular connections with host myocardial cells. The transplantation of these cells onto cryo-injured myocardium resulted in improved cardiac function in several animal models. Stem cell-derived cardiomyocytes can be selected from embryonic stem cells. These cells still divide and differentiate into different cardiac muscle cells (atrial, ventricular, and pacemaker). After transplantation into damaged myocardium in mice, they formed stable grafts and survived for at least 7 weeks. The selection of these cells has to be performed with care to prevent teratoma formation originating from single undifferentiated cells attached to the transferred cells. Recent experimental studies revealed the ability of bone marrow stem cells to differentiate into cardiomyocytes. These cells were transplanted into damaged myocardium via coronary perfusion. They survived for at least 4 weeks and showed differentiation toward cardiac muscle cells. The functional benefit of bone marrow stem cells, however, has not been clearly demonstrated, and there is a possibility of tumor formations originating from these cells. DISCUSSION: Myoblasts and bone marrow stem cell-derived cardiomyocytes would permit autologous cell transfer onto the myocardium. These cells can be easily obtained and expanded in culture. Gene transfer is also possible, and there are no ethical proscriptions against an autologous cell transfer. However, the integration and final differentiation of these cells in the heart tissue is not clear yet. Fetal cardiomyocytes, on the other hand, are integrated in the myocardial tissue, improve cardiac function, and can be expanded in culture. Their transfer would be allogenic, making immunosuppression necessary. Stem cell-derived cardiomyocytes could be used to replace all 3 types of cardiac muscle cells, and they can be expanded in culture. The possibility of teratoma formation makes a 100%- selection mandatory. At present, ethical concerns against working with human embryonic stem cells are a factor to be considered. CONCLUSIONS: Cell transfer therapy has been shown to improve myocardial function in animal experiments. This finding indicates that a reduced myocardial function can be improved by cell transfer therapy. Stem cell-derived cardiomyocytes in particular, either of embryonic or bone marrow cell origin, would allow for selective replacement of pacemaker cells or atrial or ventricular cardiomyocytes. PMID- 12538128 TI - An incidental finding of thymic carcinoid during urgent CABG operation. AB - We present a case report of a 60-year-old Malay man who was undergoing an urgent coronary artery bypass graft (CABG) operation when a well encapsulated thymoma like tumor was found incidentally. Total thymectomy was performed together with the CABG. Histopathological report of the tumor, however, confirmed a rare thymic carcinoid. The clinical features, management, and outcome of surgery are discussed. PMID- 12538129 TI - HIV-related aneurysm of the aortic root in a patient outside Africa: a case report. AB - We report a patient infected with human immunodeficiency virus (HIV) with an aneurysm of the aortic root. No definitive histopathological diagnosis was entertained in this case. Based on the existing literature, we discuss a potential causative relationship between HIV and aortic aneurysm. PMID- 12538130 TI - Ascending aortic dissection without intimal tear: a case report. AB - Aortic dissection may occur without the presence of intimal tear, and it may occur with medial dissection and intramural hematoma. We report a case in which mediastinal enlargement was found in the chest x-ray of a 79-year-old patient with chest and back pain that had started suddenly 1 week before. The patient had a decrease in hematocrit, and transthoracic echocardiography revealed around the heart pericardial fluid 5 cm thick. The ascending aorta could not be evaluated because of the presence of this fluid. The preoperative diagnosis, based on the computerized tomography findings (dissection of ascending aorta and pericardial fluid), was ruptured dissection of the ascending aorta. The patient underwent an emergency operation. Two liters of hemorrhagic fluid was aspirated from the pericardium during the operation. The ascending aorta was opened, but there was no intimal tear. Medial dissection and intramural hemorrhage were seen. The ascending aorta was replaced with a tube graft. Cases such as this, of medial dissection and intramural hematoma in which intimal integrity is preserved, should be approached in the same manner as classical dissections with intimal tear. PMID- 12538131 TI - Total Myocardial Revascularization Fifth International Symposium: an introduction. PMID- 12538132 TI - Aiming towards complete myocardial revascularization without cardiopulmonary bypass: a systematic approach. AB - BACKGROUND: Coronary artery bypass grafting (CABG) has become the surgical procedure of choice for symptomatic coronary artery disease. However, the use of traditional cardiopulmonary bypass (CPB) techniques represents an invasive therapeutic system with immediate and long-term complications. Off-pump myocardial revascularization has emerged as an attractive alternative that offers improvements in early outcomes and avoidance of the recognized adverse affects of CPB. A major criticism of this procedure has been a perceived inability to accomplish complete revascularization of the heart. In this report, we describe a surgical technique we have used in a series of patients that has allowed complete myocardial revascularization. METHODS: Combinations of intraoperative techniques were employed, including (1) right pleural-pericardial window, (2) deep pericardial sutures, (3) right heart displacement, (4) intermittent hypotensive anesthesia, (5) multimodality brain monitoring, and (6) coronary shunting. Following surgery, coronary artery grafts performed were statistically compared to each coronary artery's vascular territory to show that all territories were equally treatable with the combination of techniques. RESULTS: There were 734 coronary artery grafts performed in 200 consecutive patients (mean of 3.7 grafts/patient), and 533 compromised vascular territories were revascularized (mean of 1.38 grafts for each diseased vessel). Eight patients had one-vessel disease, 51 had two-vessel disease and 141 had three-vessel disease. The left anterior descending coronary artery (LAD) was compromised in 192 patients, the circumflex in 171 and the right coronary artery in 170 patients. The overall 30 day estimated hospital mortality was 5.5%; the observed was 4.0% (8 of 200). Postoperative complications included pulmonary insufficiency in 6 patients (3.0%), reoperation for bleeding in 3 patients (1.5%), cerebrovascular accident in 3 patients (1.5%), renal dysfunction in 2 patients (1.0%), perioperative myocardial infarction in 8 patients (4.0%), cardiac arrest in 2 patients (1.0%), low cardiac output in 5 patients (2.5%), and deep sternal infection in 2 patients (1.0%). CONCLUSIONS: Use of intermittent hypotensive anesthesia in conjunction with multimodality brain monitoring, right heart displacement, deep pericardial sutures, coronary shunting and epicardial compression stabilization facilitates complete revascularization of the myocardium. PMID- 12538133 TI - Hemodynamic changes during cardiac manipulation in off-CPB surgery: relevance in brain perfusion. AB - The recent introduction of various cardiac stabilization and positioning devices, alone or in combination with deep pericardial traction sutures, has greatly increased the ability to perform beating heart surgery to accomplish multi-vessel coronary revascularization without the need for cardiopulmonary bypass (CPB), with its associated risks. However, positioning the heart for anastomosis of the circumflex (Cx) and the posterior descending artery poses a risk of inducing hypotension, impaired cardiac output, and generalized hemodynamic instability with risk of cerebral compromise. This report discusses clinical studies suggesting that compromised right ventricular diastolic filling as a result of direct ventricular compression, rather than impaired contractility or ischemia, may be the primary mechanism for producing hemodynamic instability during OPCAB surgery. Foremost among measures to minimize ventricular compression is optimal placement of the myocardial stabilization device. Secondary measures include steep Trendelenburg positioning, fluid loading, right-sided pleuro-pericardial window that allows rotation of the heart by partial herniation into the right pleural cavity, and possibly certain pharmacological agents. This report also analyzes the effect that variable degrees of hemodynamic disturbance accompanying displacement of the heart for OPCAB surgery has on endorgan perfusion and considers the effects of hypotensive agents, direct cerebral dilators, and patient-specific factors on cerebral blood flow. The role of the partial aortic occlusion clamp and risk of stroke is also considered. We conclude that for cardiac surgery patients considered at increased risk of adverse central nervous system events, direct monitoring of cerebral function and avoidance of aortic manipulation is strongly recommended. PMID- 12538134 TI - Multi-modality neurophysiologic monitoring for cardiac surgery. AB - BACKGROUND: A high percentage of patients who undergo cardiac surgery experience persistent cognitive decline. The costs to insurers from brain injury associated with cardiac surgery is enormous. Furthermore, the same processes that injure the brain also appear to cause dysfunction of other vital organs. Therefore, there are great clinical and economic incentives to improve brain protection during cardiac surgery. This article discusses the methods of monitoring neurophysiologic function during heart surgery, including electroencephalography (EEG), near-infrared spectroscopy (NIRS), transcranial doppler (TCD) ultrasound, and cerebral oximetry, and analyzes the effectiveness of multi-modality neuromonitoring. METHODS: Neurophysiologic studies have implicated hypoperfusion and dysoxygenation as major causative factors for brain injury during cardiac surgery. Since these functional disturbances are often detectable and correctable, there is a new impetus to examine the role of neurophysiologic monitoring in brain protection. We have used a retrospective, single-surgeon case control study to examine the influence on outcome following myocardial revascularization of multi-modality neuromonitoring, with modalities that include 4-channel EEG, bilateral cerebral oximetry, and single channel TCD. RESULTS: The majority of noteworthy functional disturbances detected by neuromonitoring can be corrected by simple adjustments in perfusion, oxygenation, or anesthetic administration. In more recalcitrant cases, pharmacological neuroprotection has proven effective. In addition to the substantial reductions in length of hospital stay, costs, and neurologic complications, the results of neuromonitoring suggest a possible benefit to other vital organ systems. Future studies of neuromonitoring efficacy should not overlook these important accessory benefits. CONCLUSION: This study provides the clearest evidence to date that multi-modality neuromonitoring for cardiac surgery is safe, clinically beneficial, and cost effective. Although neuromonitoring involves negligible risk and modest costs, it's benefits for patient outcome and cost control are substantial. PMID- 12538135 TI - Patient selection in off-cardiopulmonary bypass revascularization. AB - BACKGROUND: The last few years have been marked by striking advances in off-pump coronary artery bypass grafting (OPCABG), thanks mainly to developments in instrumentation and technical progress in the revascularization of the posterior arteries of the heart. In 2001, OPCABG was performed on approximately 20% of patients who underwent surgical myocardial revascularization in the United States. This retrospective study aims to evaluate our experience with this procedure over the past 10 years, emphasizing the rapid and increasing progress of the method, indications, contraindications, and results. METHODS: Between August 1991 and December 2001, 2,759 patients suffering from angina pectoris underwent OPCABG. The patients' ages ranged from 13 to 93 years with a mean of 63 years +/- 12 years. The over 80-year-old group accounted for 3.5% of the patients. Males represented 58% of the patients. RESULTS: Intraoperative mortality in OPCABG was low. In the intraoperative period 0.4% of patients died, and the figure for the postoperative period was 2.5%. It was noted that 7.9% of the patients presented serious, nonfatal complications. CONCLUSIONS: OPCABG is a reproducible procedure with results similar to those obtained in revascularization of the myocardium using extracorporeal circulation (ECC). We believe that over 90% of patients can be operated upon without the aid of extracorporeal circulation (ECC). In our experience, all patients are potential candidates for myocardial revascularization surgery without ECC. PMID- 12538136 TI - Transit time flow measurements in coronary surgery: the experience from a new center in Bosnia. AB - BACKGROUND: A new cardiovascular center in Tuzla, Bosnia Herzegovina was opened for cardiac procedures in September 1998. In the first three years of operation, a total of 440 coronary artery bypass grafting (CABG) procedures were performed there. Off-pump coronary artery bypass (OPCAB) was emphasized as the main tool for surgical revascularization. Transit time flow measurement (TTFM) was used routinely to check graft patency. The purpose of this paper is to report on flowmetry results in the Tuzla CABG population. METHODS: All patients were considered candidates for both on-pump (ONCAB) and off-pump (OPCAB) CABG procedures. Approximately 60% of the procedures were performed as ONCAB and the rest as OPCAB. For all patients, TTFM was performed on all grafts. RESULTS: Eighteen patients were converted from OPCAB to ONCAB. Revision was required for 1.8% of the grafts. All grafts were successfully revised and were patent at the time of wound closure. CONCLUSION: We believe that TTFM is a crucial tool in CABG. It offers a reliable and inexpensive tool for quality assurance in coronary revascularization. PMID- 12538137 TI - How to start a beating heart coronary artery surgery program. AB - An important issue that is not addressed in the recent literature on beating heart coronary artery bypass surgery is the question of how to initiate a program in OPCAB in a center where conventional coronary artery surgery has been the routine. It is the aim of this article to describe what we consider to be safe steps for a conventional cardiac surgeon to take to ensure a successful initiation of OPCAB surgery in his or her institution. While modern OPCAB surgery has experienced major technical advances that have made the procedure more predictably successful than in previous years, these advances require the use of an array of new technology and skills. This article discusses some of the skills and devices, such as modern stabilizers, shunts, and blowers, that the beginning OPCAB surgery team will need to acquire and master when embarking on an OPCAB program. Initiation of an OPCAB program also requires that the entire surgery team believe in the efficacy of the procedure and approach it with enthusiasm. A team visit to a center that already performs OPCAB is an important element in initiating the new program. It is no longer necessary to learn the procedures by trial and error. Because it will occasionally be necessary to convert from OPCAB to conventional surgery, the team must be thoroughly familiar with both methods and should not be reluctant to rely on the heart-lung machine when the patient's safety requires it. PMID- 12538138 TI - Off-pump coronary artery bypass surgery--initial experience in Gdansk: a brief review. AB - BACKGROUND: Before the introduction of cardiopulmonary bypass (CPB), coronary artery bypass grafting (CABG) procedures were performed on a beating heart. In 1967, Kolessov first reported "off-pump" revascularization of the left anterior descending artery (LAD). This technique was later abandoned when the use of cardioplegia and the heart-lung machine allowed a motionless and bloodless operative field. This study reports our initial clinical experience in off-pump coronary bypass surgery performed at the Department of Cardiac Surgery of the Medical University of Gdansk, Poland. METHODS: This study enrolled all consecutive patients who were operated on at our institution without CPB between January 1998 and December 2001. Patients were selected for the off-pump procedure individually by the surgeon. Demographics, operative procedures, postoperative mortality, morbidity, and early outcomes were analyzed. The observation period included 30 postoperative days. RESULTS: An average of 1.8 grafts per patient were completed. Conversion to CPB was required in 4.1% of patients. Complete revascularization was attained in 91% of the procedures. Surgical mortality was 1.3% and perioperative myocardial infarction was reported in 2.4%. CONCLUSIONS: In selected patients, off-pump CABG may be used as a suitable and safe alternative to conventional on-pump coronary surgery and permits complete revascularization with comparable short-term results. PMID- 12538140 TI - Emboli, inflammation, and CNS impairment: an overview. AB - Perioperative stroke occurs in 2-3% of adult cardiac surgery patients, and significant cognitive dysfunction is experienced by 40-60% of patients in the first postoperative week. Perioperative neurocognitive abnormalities are associated with a greatly increased risk of perioperative mortality, lengthy intensive care and hospital stay, and more intensive rehabilitative care. Long term cognitive dysfunction, ranging from months to years, occurs in 25-40% of adult cardiac surgery patients, resulting in a decreased quality of life. Cerebral emboli are an important cause of perioperative neurocognitive abnormalities. Aortic cannulation, clamping, and manipulation during surgery may dislodge atheromatous materials into the cerebral circulation, leading to perioperative or postoperative stroke. Nevertheless, acute and chronic neurocognitive dysfunction frequently occurs in non-cardiac surgery patients as well, suggesting that some element of surgery and/or anesthesia itself causes or contributes to this phenomenon. One possible cause may be central nervous system (CNS) responses to peripheral tissue injury or inflammation. The CNS is sensitive to systemic pro-inflammatory mediators such as endotoxin and the cytokines interleukin-6 and interleukin- 8, which are activated by surgical trauma. This article discusses the behavior and effects of these inflammatory agents and their intensification in combination with postoperative hyperthermia. The potential beneficial role of pharmacological agents such as heparin, lidocaine, and aprotinin is also examined. PMID- 12538141 TI - Normal accidents: human error and medical equipment design. AB - High-risk systems, which are typical of our technologically complex era, include not just nuclear power plants but also hospitals, anesthesia systems, and the practice of medicine and perfusion. In high-risk systems, no matter how effective safety devices are, some types of accidents are inevitable because the system's complexity leads to multiple and unexpected interactions. It is important for healthcare providers to apply a risk assessment and management process to decisions involving new equipment and procedures or staffing matters in order to minimize the residual risks of latent errors, which are amenable to correction because of the large window of opportunity for their detection. This article provides an introduction to basic risk management and error theory principles and examines ways in which they can be applied to reduce and mitigate the inevitable human errors that accompany high-risk systems. The article also discusses "human factor engineering" (HFE), the process which is used to design equipment/ human interfaces in order to mitigate design errors. The HFE process involves interaction between designers and endusers to produce a series of continuous refinements that are incorporated into the final product. The article also examines common design problems encountered in the operating room that may predispose operators to commit errors resulting in harm to the patient. While recognizing that errors and accidents are unavoidable, organizations that function within a high-risk system must adopt a "safety culture" that anticipates problems and acts aggressively through an anonymous, "blameless" reporting mechanism to resolve them. We must continuously examine and improve the design of equipment and procedures, personnel, supplies and materials, and the environment in which we work to reduce error and minimize its effects. Healthcare providers must take a leading role in the day-to-day management of the "Perioperative System" and be a role model in promoting a culture of safety in their organizations. PMID- 12538142 TI - Using the STS and multinational cardiac surgical databases to establish risk adjusted benchmarks for clinical outcomes. AB - One of the purposes of collecting data on cardiac surgical procedures, at a national level is to enable individual surgeons to improve quality and benchmark their own practice by making more accurate prospective prediction of outcome of each individual patient by using risk stratification based on previous local and national experiences. The past decade has seen a dramatic increase in the development of national cardiac surgical initiatives in many countries around the world. The size and extent of these databases has successfully allowed their use for patient risk stratification and preoperative risk modeling in four main aspects: patient selection and informed consent, coherent analysis of the determinants of patient outcomes, rationalizing unit management, and negotiations with external agencies. Approximately 610 cardiac surgical units presently contribute their patient data, containing pre-operative risk factors, to centralized national registries. There are currently nine different datasets used throughout the world to collect patient information. To harmonize the considerable diversity among these source materials, an International Dataset has been developed by a collaborative process among more than 50 cardiac surgeons around the world. Constructed around the Society of Thoracic Surgeons (STS) data format, the International Dataset brings in key elements from all the other datasets, allowing the sharing of data and cross-analysis, thus greatly expanding the pool of patients, and national sources, from which risk-stratifed outcomes can now be analyzed and unified. Unlike the STS dataset, the International Dataset incorporates EuroSCORE, a simple-to-use, validated patient risk stratification system, which has been rapidly adopted by large numbers of centers around the world for patient risk stratification, outcomes assessment, and improving patient informed consent. There are several benefits to collecting and centralizing national and international data: (1) understanding and defining basic demographics of patients undergoing cardiac surgery; (2) patient risk stratification and risk prediction at both a national and center-by-center level; (3) unit benchmarking, and development of effective nationally oriented and center-oriented quality improvement programs; (4) understanding and rationalizing resource utilization; and (5) use of data to leverage governments and other healthcare providers to affect policy. Cardiac surgical registries will soon attempt to track patients for longer follow-up periods after discharge in order to identify surgery-related deaths for more extended periods of time following surgery, thereby improving the monitoring and prediction of patient outcomes. PMID- 12538143 TI - Cardiac positioning using an apical suction device maintains beating heart hemodynamics. AB - BACKGROUND: Cardiac positioning during off-pump coronary artery bypass (OPCAB) using deep pericardial sutures (DPS) typically results in some degree of hemodynamic compromise. We sought to determine whether cardiac positioning using an apical suction device was hemodynamically superior to DPS. METHODS: Five healthy pigs underwent sternotomy and instrumentation to measure right atrial (RA) pressure, left ventricular (LV) pressure and volume, and aortic pressure and flow. These variables were recorded at baseline, with simple attachment of the apical suction device (Xpose Access Device, Guidant, Inc.), and during exposure of the posterior descending artery (PDA) and obtuse marginal (OM) branches of the left circumflex artery using DPS and the apical suction device. RESULTS: Application of the apical suction device to the beating heart in neutral anatomic position did not result in any statistically significant change in hemodynamics compared to baseline except for a small decrease in RA pressure. DPS positioning resulted in statistically significant compromise in nearly all measured hemodynamic parameters, including cardiac output (-21% PDA, -30% OM), mean arterial pressure (-18% PDA, -26% OM), and stroke work (-31% PDA, -38% OM). In addition, LV end-diastolic pressure decreased (-59% PDA, -51% OM) while RA pressure increased (+17% PDA, +16% OM). Similar target exposure using the apical suction device resulted in near-baseline hemodynamics. The only statistically significant changes were a modest decrease in cardiac output (-18% OM) and RA pressure (-11% PDA). CONCLUSION: DPS positioning significantly compromises hemodynamics due to reduced LV filling. The apical suction device provides good exposure with less hemodynamic compromise. PMID- 12538144 TI - Vacuum-assisted venous drainage does not increase the neurological risk. AB - BACKGROUND: Vacuum-assisted venous drainage (VAVD) with negative pressure applied to integral sealed-hardshell venous reservoir facilitates valvular surgery through minimally invasive approaches. Despite concerns regarding air entrainment from the right atrium, cerebral microemboli of air and neurological complications, VAVD was used in patients who underwent valvular surgery throughout the last two years in our institution. METHODS: We compared the rate of neurological complications in patients who underwent surgery with and without VAVD from June 1997 to July 2001. VAVD was added to solid venous reservoirs with membrane oxygenators and arterial filters. Clinical results were prospectively entered in our valve database and were used for the analysis. RESULTS: Eight hundred twenty-two consecutive patients averaging 65 +/- 11 years of age underwent aortic, mitral and tricuspid valve replacements including 40 redos (40/822, 5%) and 265 associated CABG (265/822, 32%) with VAVD in 1999 to 2001 compared to 723 patients averaging 63 +/- 11 years of age (p = 0.01) who underwent the same procedures with 79 redos (79/723, 11%) and 177 CABG (177/723, 24%) without VAVD in 1997 to 1999. CPB time averaged 117 +/- 50 minutes in VAVD patients compared to 108 +/- 43 minutes in those without VAVD (p = 0.001). Thirty day mortality averaged 5% (39/822) in patients with VAVD and 4% (30/723) in those without VAVD (p = 0.6). Seven patients of the VAVD group (7/822, 1%) and 11 patients without VAVD (11/723, 1.5%, p = 0.2) suffered from temporary or permanent neurological deficit. CONCLUSION: VAVD is a useful adjunct to modern cardiopulmonary bypass systems. When used with appropriate care, VAVD does not appear to significantly increase air microemboli and is not associated with an increased neurological risk following valvular surgery. PMID- 12538145 TI - Optimizing revascularization of complex, proximal left anterior descending coronary stenoses: the clinical and financial impacts of catheter-based, median sternotomy, and port-access approaches. PMID- 12538146 TI - Totally endoscopic atrial septal defect repair with robotic assistance. AB - BACKGROUND: The development of minimally invasive cardiac surgery has been characterized by the performance of increasingly complex operations through progressively smaller incisions. Computer (robotic) enhancement has emerged as a potential facilitator of these procedures, initially by providing enhanced endoscopic camera control and, more recently, by allowing the manipulation of surgical instruments through limited thoracic incisions. This report describes the next step in this progression, namely the performance of an atrial septal defect (ASD) repair entirely through thoracoscopic port incisions. This represents the first U.S. application of robotic technology for totally endoscopic open-heart surgery. MATERIALS AND METHODS: A 33-year-old woman with a secundum atrial septal defect underwent totally endoscopic repair through four port incisions by means of the Da Vinci (Intuitive Surgical, Mountain View, CA) robotic surgical system. Cardiopulmonary bypass was achieved peripherally (femoral Estech endoaortic balloon cannula; femoral and right internal jugular venous Bio-medicus cannulae). The myocardium was protected with antegrade cold blood cardioplegia delivered through the distal port of the arterial cannula. After port insertion, the entire operation, including pericardiotomy, bicaval occlusion, atriotomy, atrial septopexy, and atrial closure, was performed by computer-aided control of a camera and two instrument arms manipulated by a surgeon seated 15 feet away. The fourth port was used for suction and suture passage by the patient-side assistant. The aortic cross-clamp time was 43 minutes, and the postoperative transesophageal echocardiogram demonstrated normal ventricular function and the absence of interatrial shunting. The patient was extubated on the night of surgery, was ambulatory within 15 hours, and was discharged on the morning of postoperative day 3, 63 hours after the procedure. At 30-day follow-up, the patient was well and without complaints, and transthoracic echocardiogram confirmed the continued absence of interatrial shunting. CONCLUSIONS: Computer-aided robotic surgical technology can be used to perform open-heart procedures with a totally endoscopic approach. The benefits of this approach may include decreased perioperative pain, decreased recovery times, and improved cosmesis and patient acceptance. Clinical trials currently in progress will demonstrate whether this technology will be of reproducible value in the management of patients with intracardiac disease on a larger scale. PMID- 12538147 TI - Mitral valve aneurysm associated with aortic valve endocarditis and regurgitation. AB - Mitral valve aneurysms are rare complications occurring most commonly in association with aortic valve infective endocarditis. [Decroly 1989, Chua 1990, Northridge 1991, Karalis 1992, Roguin 1996, Mollod 1997, Vilacosta 1997, Cai 1999, Vilacosta 1999, Teskey 1999, Chan 2000, Goh 2000, Marcos- Alberca 2000] While the mechanism of the development of this lesion is unclear, complications such as perforation can occur and lead to significant mitral regurgitation. [Decroly 1989, Karalis 1992, Teskey 1999, Vilacosta 1999]; The case of a 69-year old male with Streptococcus Sanguis aortic valve endocarditis and associated anterior mitral leaflet aneurysm is presented. Following surgery, tissue pathology of the excised lesion revealed myxomatous degeneration and no active endocarditis or inflammatory cells. This may add support to the hypothesis that physical stress due to severe aortic insufficiency and structural weakening, without infection of the anterior mitral leaflet, can lead to the development of this lesion. PMID- 12538148 TI - Anomalous course of the right coronary artery in the right atrial wall: a word of caution. AB - We report a case of anomalous course of the right coronary artery in the wall of right atrium which was encountered during coronary artery bypass surgery. As the stenotic lesion in the vessel was proximal, the large posterior descending branch of the right coronary was grafted. Such an anomalous course of the right coronary artery has not been previously described in the literature and lack of knowledge of such an abnormal course may result in inadvertant damage during cannulation of the inferior vena cava or coronary sinus. PMID- 12538150 TI - How microarrays can improve our understanding of immune responses and vaccine development. PMID- 12538151 TI - Historical background and anticipated developments. AB - Expression profiling using DNA arrays is often believed to have appeared during the second half of the 1990s, and to be based exclusively on nonisotopic methods. In fact, the first article describing the application of cDNA arrays to expression analysis was published in 1992, relied on radioactive labeling, and was a new development of "high-density" membranes used until then essentially for efficient screening of libraries. Several papers described the use of this technology for simultaneous expression measurement of thousands of genes at the time when the first glass microarrays were published. Simultaneously, oligonucleotide chips, originally developed for resequencing and mutation detection applications, were shown to be capable of expression measurement as well. The three approaches have developed over the years and still coexist, as each of them has specific advantages (and drawbacks); the major issues have become those of data quality, data analysis and storage (ideally in a common public database). Meanwhile, the technology continues to evolve. The most obvious trend is a shift towards using arrays of relatively long oligonucleotides that combine most of the advantages of very long (cDNA) and very short (25-mer) DNA segments. The search for better detection methods, ideally without labeling of the sample, is continuing, although it seems difficult to reach the required sensitivity. New materials for microarray manufacture and new implementations of existing methods have appeared. In addition, the field is progressively becoming segmented into high gene number, low volume (research) applications on the one hand, and low gene number, high throughput (diagnostic) uses on the other. PMID- 12538152 TI - Analysis of B cell memory formation using DNA microarrays. AB - DNA microarray analysis of B cell subsets has identified comprehensive programs of gene expression that distinguish B cells at discrete stages of differentiation. The next task is to identify key genetic signals within these complex programs that regulate the dynamic cellular events during B cell activation in vivo. After stimulation with antigen, naive B cells proliferate and differentiate, and then produce antibodies. Crucial qualitative differences in antibody responses are observed depending on whether or not B cells receive T cell help during activation. Proteins, lipopolysaccharides, and polysaccharides stimulate T-dependent (TD), T-independent type 1 (TI-1), and type 2 (TI-2) antibody responses, respectively. Only TD responses generate somatically mutated antibody-forming (plasma) cells and memory B cells, which produce high affinity anamnestic responses to subsequent antigen challenge. Somatic mutation of immunoglobulin genes occurs during B cell proliferation in germinal centres (GC), which are typical in TD responses but rare in TI responses. However, we have described a model, which is exceptional because numerous large GC form in response to a model TI-2 antigen, (4-hydoxy-3-nitrophenyl) acetyl (NP)-Ficoll. Significantly, these GC undergo involution before memory B cells are generated. This model provides an opportunity to investigate the genetic signals that drive memory cell formation, and we have compared global gene expression in TI and TD GC to identify a relatively small number of genes that are differentially expressed between the two prototypic B cell responses. This model demonstrates how genome-scale technology can be adapted to investigate specific aspects of B cell biology. PMID- 12538154 TI - A genomic view of helper T cell subsets. AB - Genomic-scale gene expression profiling in combination with the availability of a draft sequence of the human genome is beginning to revolutionize the way immunology is done. The possibility of measuring levels of gene expression for tens of thousands of genes simultaneously and in a quantitative fashion aids in the definition of a comprehensive molecular phenotype of cells and cellular processes of the immune system in health and disease. T helper lymphocytes are an essential element of appropriate immune responses to pathogens. To achieve effective immunity, T helper cells differentiate into at least two specialized subsets that direct type 1 and type 2 immune responses. Here, I discuss recent progress that has been made in our understanding of the genetic program that controls the development and functional properties of helper T cell subsets. PMID- 12538153 TI - IL-15 is a growth factor and an activator of CD8 memory T cells. AB - Memory lymphocytes, arising from naive lymphocytes after antigenic stimulation and being long-lived, are the cellular basis for immunological memory. Recent studies of CD8 T cells suggest that generation of CD8 memory T cells requires the engagement of T cell antigen receptors (TCR) with antigen, yet the maintenance of CD8 memory T cells appears to be dependent on cytokines, such as IL-15, independent of TCR. Although considerable progress has been made in understanding the molecular and cellular events of TCR-induced differentiation and proliferation in the past decade, less is known about the mechanisms of IL-15 action. From a kinetic and comparative analysis of the responses of memory phenotype CD8 T cells to IL-15 and TCR stimulation in vitro, we found that IL-15 and anti-CD3 induce highly similar responses in memory phenotype CD8 T cells as measured by general gene expression profiles, synthesis of effector molecules (IFNgamma, TNFbeta, granzyme B and perforin), induction of cytotoxicity, and cellular proliferation. These findings indicate that IL-15 is not only a growth factor but also an antigen-independent activator for CD8 memory T cells. PMID- 12538155 TI - Gene profiling approach to establish the molecular bases for partial versus full activation of naive CD8 T lymphocytes. AB - When initial antigen encounter involves optimal antigenic and costimulatory stimuli, naive CD8 T cells undergo a developmental program that leads to their activation, expansion and acquisition of effector functions (including production of IL-2, IFNgamma and expression of cytolytic effector molecules). A subset of the activated CD8 T cells thrives as long-lived memory cells. Encounter of tissue associated, and in particular tumor-associated antigen, may often be suboptimal in terms of antigenicity and costimulation, however. We previously developed a model of naive CD8 T cells from transgenic mice expressing an alloreactive TCR for which a mutant alloantigen behaved as a partial agonist, inducing only some of the effector functions induced by the native alloantigen. To ascertain the molecular bases for the establishment of divergent fates within the same naive CD8 T cells, we have used cDNA microarrays to monitor sequential gene expression patterns in conditions of full or partial response of these naive CD8 T cells. Of the 5000 different genes monitored on the array, 18% showed changes in expression in activated versus naive CD8 T cells, independent of whether stimulation was with full or partial agonist. These included antigen-induced upregulated as well as downregulated genes. Clusters of genes that were differentially expressed were also identified, being either (i) weakly versus strongly, or (ii) transiently versus stably expressed in response to partial and full agonist, respectively. They included (i) genes encoding costimulatory molecules and (ii) genes controlling cytolytic function, cytokine production, and chemokines. Therefore, the cDNA microarray approach was a sensitive tool to provide an exhaustive picture of T cell activation as it could discriminate quantitative, qualitative and dynamic differences in mRNA expression profiles between fully or partially activated T cells. PMID- 12538156 TI - Complexity of inflammatory responses in endothelial cells and vascular smooth muscle cells determined by microarray analysis. AB - To better understand the molecular basis of vascular cell system behavior in inflammation, we used gene expression microarrays to analyze the expression of 7,075 genes and their response to IL-1beta and TNFalpha in cultures of coronary artery endothelium and smooth muscle derived from a single coronary artery. The most noticeable difference between the cell types was the considerably greater magnitude and complexity of the transcriptional response in the endothelial cells. Two hundred and nine genes were regulated in the endothelium and only 39 in vascular smooth muscle. Among the 209 regulated genes in the endothelium, 99 have not been previously associated with endothelial cell activation and many implicate the endothelium in unconventional roles. For example, the induced genes include several that have only been associated with leukocyte function (e.g., IL 7 receptor, EBI-3 receptor) and others related to antiviral and antibacterial defense (e.g., oligoadenylate synthetase, LMP7, toll-like receptor 4, complement component 3). In addition, 43 genes likely to participate in signal transduction (eg. IL-18 receptor, STK2 kinase, STAF50, ANP receptor, VIP receptor, RAC3, IFP35) were regulated providing evidence that a major effect of TNFalpha and IL 1beta is to alter the potential of the endothelial cell to respond to various other external stimuli. PMID- 12538157 TI - Integrated genomic and proteomic analysis of signaling pathways in dendritic cell differentiation and maturation. AB - Dendritic cells (DCs) are antigen-presenting cells that play a major role in initiating primary immune responses. Their phenotypic and functional characteristics are intimately linked to their stage of maturation. The specific biochemical pathways and genes whose expression mediates differentiation of progenitors to DCs and their maturation are largely undefined. We recently utilized two approaches, DNA microarrays and proteomics, to analyze the expression profile of human CD14(+) blood monocytes and their derived DCs. Approximately 4% of the genes or proteins expressed were found to be regulated during DC differentiation. Most of these genes were not previously associated with DCs and included genes highly relevant to DC functions (genes involved in antigen presentation, cell adhesion and motility, lipid metabolism). Genes involved in specific signaling pathways, including IkappaBalpha, PPAR-gamma and C/EBPalpha as well as two members of the family of transcription factors, interferon regulatory factors (IRFs), were also modified. Modulation of IRF gene expression is of particular interest because of their functional roles in innate and adaptive immune responses. IRF-family members control the expression of proteins that include type-1 interferons, interleukin-12, interleukin-15, MHC molecules and adhesion molecules. They have also been found to play an important role in lymphocyte development. In contrast to DC differentiation, very few genes were modified at the transcript level during DC maturation as determined by microarray experiments. Further analysis suggested that DC maturation is largely controlled by posttranscriptional and posttranslational modifications. The use of proteomics is therefore necessary for a full comprehension of DC maturation process. PMID- 12538158 TI - CD30-governor of memory T cells? AB - CD30 is well recognized as a marker expressed by a heterogeneous group of lymphomas and in several immune and autoimmune disorders. However, the function of CD30 in theses diseases or in the normal immune response has remained unclear. Studying gene expression patterns induced by stimulating CD30 signals on a large granular lymphoma cell line, YT, with an agonistic anti-CD30 antibody, we found that CD30 signals affected proapoptotic and antiapoptotic genes, regulated cytotoxicity, and controlled molecules regulating T cell traffic. Creating CD30-L deficient mice and studying CD8 CTL activation and memory responses, it was observed that the absence of CD30-L resulted in diminished primary clonal expansion of CD8 cells. In addition the absence of CD30-L abolished clonal contraction after primary expansion and interfered with secondary expansion upon boosting. The studies suggest that CD30 regulates CD8 CTL function and survival during memory responses and is important for clonal contraction. PMID- 12538159 TI - Stronger correlation of bcl-3 than bcl-2, bcl-xL, costimulation, or antioxidants with adjuvant-induced T cell survival. AB - A set of signals separate from those needed for T cell activation and clonal expansion acts to sustain a T cell response once it has begun. Immunologic adjuvants can initiate these signals in a process we designate adjuvant-induced survival (AIS). Here, the natural adjuvant LPS was used in a super-antigen model of AIS to understand which factors are needed to sustain T cell survival after activation. Flow cytometric stains for antiapoptotic Bcl-2 and Bcl-xL showed that neither factor was well correlated with AIS, although both were increased transiently upon T cell activation. T cells protected via AIS showed no increased ability to resist death caused by reactive oxygen species, and cellular division was not accelerated as might be expected if AIS were to operate through co stimulatory pathways. Finally, microarray analyses were performed that showed increased expression of Bcl-3, an NFkappaB/IkappaB factor, was correlated with AIS. It is proposed that T cell survival during productive immune responses occurs by successive activities of Bcl-2, Bcl-xL and Bcl-3, with Bcl-3 requiring innate immune responses to adjuvants for its expression. PMID- 12538160 TI - A short domain within Bcl-3 is responsible for its lymphocyte survival activity. AB - The NFkappaB factor Bcl-3 influences the survival of T cells when they are activated to take part in immune responses. Because treatment of mice with adjuvant results in the increased expression of Bcl-3 in T cells, where it has survival-promoting effects, Bcl-3 may be an important, limiting factor that is supplied to T cells only when they are contributing to an appropriate immune response to infection, and not when spuriously activated by self-antigens. Although Bcl-3 is a member of the NFkappaB/Rel/IkappaB family of transcription factors, the means by which it promotes T cell survival is not obvious because Bcl-3 is unique in having an ankyrin repeat domain, like inhibitory IkappaB proteins, while also possessing domains capable of transcriptional activation, like Rel proteins. In order to understand the basis for the survival activity of Bcl-3, deletion mutants were engineered and tested in a retroviral gene transfer sytem. We report that most of Bcl-3 can be deleted without diminishing its ability to prolong the survival of activated T and B cells, and find that its lymphocyte survival domain maps to the vicinity of its first and second ankryin repeats. This information sets the stage for experiments in which a focused search can be made for mediators of Bcl-3 survival effects. PMID- 12538161 TI - Molecular characterization of antigen-induced lung inflammation in a murine model of asthma. AB - Asthma is one of the foremost contributors to morbidity and mortality in industrialized countries. Our objective was to characterize the acute response to allergen and to identify potentially novel molecular targets for pharmacological intervention in asthma. We therefore designed a study to identify genes whose regulation was altered following ovalbumin (OVA) challenge in the presence and absence of treatment with glucocorticoids in BALB/c mice. RNA was isolated from lungs for gene profiling from 8-week-old sensitized mice, 3 and 18 hours post OVA challenge on days 1, 4, and 7 of aerosol challenge. Taqman (real time RT-PCR) analysis of marker genes indicative of Th2 (IL-4, IL-13), eosinophil (RANTES, eotaxin), Th1/macrophage (IFNgamma) and epithelial cell (MUC5AC) phenotypes were used to characterize responses to allergen challenge. Histological evaluation of lungs from additional challenged animals revealed inflammatory infiltrates on days 4 and 7, but not on day 1 post challenge. We postulate that expression of IL 4, IL-13 and other genes by OVA at day 1 probably reflects activation of resident cells, whereas the fivefold increase in the number of regulated genes at day 7 reflects the contribution of recruited cells. Of the regulated genes, only a subset was counter-regulated by dexamethasone treatment. Although regulated genes included genes in many protein families, herein we report regulation of two proteases whose role in response to OVA challenge has not been characterized. This model will be used to generate disease hypotheses for which may play an important role in initiating disease pathology in this model. PMID- 12538162 TI - Unlocking the mysteries of virus-host interactions: does functional genomics hold the key? AB - The interactions between viruses and the cells they infect are complex and multifaceted. While viruses strive to usurp cellular functions to their advantage, the cell strives to thwart these efforts by mounting a variety of defensive responses. These responses may include the induction of interferon, stress response, or apoptotic pathways, all of which are accompanied by changes in gene expression. Some viruses consistently win this tug of war, whereas others succumb to cellular defense mechanisms. The viral and cellular factors that determine the outcome are for the most part still unknown. With the advent of functional genomics, potent new technologies are now available to probe the complexities of virus-host interactions in ever increasing depth and detail. We describe here our efforts to use microarrays, proteomics, and bioinformatics to focus in on the changes in gene expression and protein production that occur in a virus-infected cell and to use these technologies to unlock the mysteries of virus-host interactions. PMID- 12538163 TI - Genetic determinants of coxsackievirus B3 pathogenesis. AB - The development of high throughput genomic and bioinformatic analysis tools, coupled with established molecular techniques, has allowed new insights into the pathogenesis of infectious diseases. In humans, coxasackievirus B3 (CVB3) is the primary etiological agent of viral myocarditis, an inflammatory disease process involving the heart muscle. Early host cellular survival and apoptotic mechanisms during viral infections, as well as immune events, affect myocarditis progression and outcome. Therefore, our laboratory has been keenly interested in infectomics, defined here as the transcriptional events of both virus and host. We first elucidated up- or downregulated transcriptional activities in CVB3-infected hearts by mRNA differential display. Further characterization of these regulated genes including Nip21, IP10, and IGTPase, and study of their role in CVB3 infection are underway. In further dissection of the stages of myocarditis-peak viremia, inflammatory infiltration and tissue repair-we used cDNA microarrays to probe differential gene expression in the myocardium following virus infection. Following virus infection, there are global decreases in metabolic and mitochondrial genes, increases in signaling genes and distinctive patterns in other functional groups. To establish early gene expression profiles in infected cells by themselves, we also used oligonucleotide arrays in an in vitro model of CVB3 infection. Notably, we have found increased expression of transcription factors c-fos and c-jun down-stream of extracellular signal-related kinase, a pathway which is crucial for virus replication and pathogenesis. Our investigations based on gene profiling following CVB3 infection have thus far been fruitful in providing new experimental leads. High throughput genetic analysis has allowed us to simultaneously try on greater than 12,000 potential genetic "glass slippers." Our in vitro experimental plan has enabled us to chart prominent patterns of gene expression, analyzed by novel bioinformatic approaches, and to separate varied and potentially significant gene expression events. PMID- 12538164 TI - A functional genomics approach to Kaposi's sarcoma. AB - Kaposi's sarcoma (KS) is the most frequent malignancy afflicting acquired immune deficiency syndrome (AIDS) patients. Tumor lesions are characterized by spindle cells of vascular origin and vascularization. Kaposi's sarcoma-associated herpes virus (KSHV) is consistently found in all forms of KS. Infection of dermal microvascular endothelial cells (DMVEC) with KSHV recapitulates spindle cell formation in vitro. We studied this transformation process by DNA microarray analysis comparing the RNA expression profiles of KSHV-infected and mock-infected DMVEC. Genes involved in tumorigenesis, angiogenesis, host defense, cell growth and differentiation, transcription, and metabolism were observed to change significantly upon infection with KSHV. One of the most consistently KSHV-induced genes was the receptor tyrosine kinase and proto-oncogene c-Kit. Inhibition of c Kit activity with the pharmacological inhibitor of c-Kit signaling STI571 reversed the KSHV-induced morphological transformation of DMVEC. Moreover, overexpression studies showed that c-Kit was sufficient to induce spindle cell formation (Moses et al. J. Virol. 76(16): 8383-8399). These data demonstrate that microarrays are useful for the identification of pharmacological targets essential for KS tumorigenesis. PMID- 12538165 TI - Chlamydiae host cell interactions revealed using DNA microarrays. AB - Chlamydiae are obligate intracellular bacterial parasites that infect eukaryotic cells and live their entire life cycle within a cytoplasmic vacuole or inclusion. We have employed cDNA microarray and conventional biological approaches to study the pathogen-host cell interaction during C. pneumoniae infection of eukaryotic cells. Two host cell signaling pathways, MEK/ERK and PI 3-kinase/Akt, were activated within 5 and 20 minutes, respectively, following infection with chlamydiae. Pharmacological inhibition of these pathways blocked invasion of HEp2 cells indicating that activation of these pathways was required for infection. Rho family GTPase activity was essential for invasion, since the pan-Rho GTPase inhibitor, compactin, blocked infection of HEp2 cells. cDNA microarrays and reverse transcriptase PCR were used to study host cell and chlamydial gene expression during the replication cycle. Analysis of host cell gene expression following infection with C. pneumoniae indicated that genes coding for cytokines, growth factors, and signaling molecules were upregulated, as early as 2 hours postinfection. Analysis of chlamydial gene expression indicated a temporal regulation of transcription with distinct early-, mid-, and late-cycle classes of RNA transcripts. Newly discovered genes encoding three Ser/Thr protein kinases and one protein phosphatase were upregulated 6-12 hours postinfection. One protein kinase, designated CpnPK1, was first detected at 12 hours postinfection, accumulated in the inclusion throughout the replication cycle, and may be a type III effector molecule. An increased understanding of chlamydial host cell interactions, in particular the role of various chlamydial proteins in infection and identification of essential virulence factors should provide novel targets for the development of new antimicrobials. PMID- 12538166 TI - Gene expression profile in Neisseria meningitidis and Neisseria lactamica upon host-cell contact: from basic research to vaccine development. AB - Differential gene regulation in the human pathogen Neisseria meningitidis group B (MenB) and in Neisseria lactamica, a human commensal species, was studied by whole genome microarray after bacterial interaction with epithelial cells. Host cell contact induced changes in the expression of 347 and 285 genes in MenB and N. lactamica, respectively. Of these, only 167 were common to MenB and N. lactamica, suggesting that a different subset of genes is activated by pathogens and commensals. Change in gene expression was stable over time in N. lactamica, but short-lived in MenB. A large part (greater than 30%) of the regulated genes encoded proteins with unknown function. Among the known genes, those coding for pili, capsule, protein synthesis, nucleotide synthesis, cell wall metabolism, ATP synthesis, and protein folding were down-regulated in MenB. Transporters for iron, chloride and sulfate, some known virulence factors, GAPDH and the entire pathway of selenocysteine biosynthesis were upregulated. Gene expression profiling indicates that approximately 40% of the regulated genes encode putative surface-associated proteins, suggesting that upon cell contact Neisseria undergoes substantial surface remodeling. This was confirmed by FACS analysis of adhering bacteria using mouse sera against a subset of recombinant proteins. Finally, a few surface-located, adhesion-activated antigens were capable of inducing bactericidal antibodies, indicating that microarray technology can be exploited for the identification of new vaccine candidates. PMID- 12538167 TI - Prognosis of breast cancer and gene expression profiling using DNA arrays. AB - Breast cancer is a complex genetic disease characterized by the accumulation of multiple molecular alterations. The resulting clinical heterogeneity makes current therapeutic strategies-based on clinicopathlogical factors-less than perfectly adapted to each patient. Today, DNA arrays, by allowing the simultaneous and quantitative analysis of the mRNA expression levels of thousands of genes in a single assay, provide novel tools to tackle this complexity. Potential applications are multiple in the cancer field and the first research results are promising. Using home-made DNA arrays in an approach easily compatible with academic research-nylon support and radioactive detection-we identified a predictor set of 23 genes whose expression patterns differentiated two groups of breast cancer patients with different survival after adjuvant chemotherapy. We then validated and further extended these results in a larger, independent and homogeneous series of poor prognosis primary breast cancers treated with adjuvant anthracyclin-based chemotherapy. We confirmed the prognostic classification provided by the 23-gene set predictor. We then improved the predictor set and refined the classification by sorting the tumors into three classes with significantly different long-term survival. These results show the potential of the technology with an accessible approach for academic research teams. We also showed that nylon DNA arrays with radioactive detection are associated with excellent sensitivity, an advantage in clinical situations where the amount of available material is limited. PMID- 12538168 TI - Identifying immunotherapeutic targets for prostate carcinoma through the analysis of gene expression profiles. AB - Carcinoma of the prostate represents one of the most frequently diagnosed cancers in men. If detected at an early stage, prostate cancer is highly treatable. However, cancers identified at a late stage are rarely cured with contemporary medical therapies. Early detection strategies presently center on the identification of prostate-specific proteins in the serum, and emerging therapeutics have utilized genes and proteins with prostate-restricted expression for tissue-selective immunological regimens incorporating vaccines, dendritic cell therapy, gene therapy, and antibody-based cell targeting. In order to develop improved therapeutic procedures, efforts have been directed toward the identification of genes exhibiting prostate-restricted expression profiles, or altered expression levels in neoplastic cells relative to their normal counterparts. Comprehensive expression profiling approaches such as the analysis of oligonucleotide- or complementary DNA (cDNA)-microarrays have greatly enhanced these efforts. Genes and their cognate proteins identified using such methods offer additional diagnostic and therapeutic targets that may aid in the understanding and treatment of prostate carcinoma. PMID- 12538169 TI - The large cost of critical care: realities and challenges. PMID- 12538170 TI - When it comes to outcome, we need to define what a perioperative epidural technique is. PMID- 12538171 TI - Missing the informed in consent. PMID- 12538172 TI - Intermittent or continuous carbon dioxide insufflation for de-airing of the cardiothoracic wound cavity? An experimental study with a new gas-diffuser. AB - Insufflation of carbon dioxide into the chest wound is used in open-heart surgery to de-air the heart and great vessels. In a cardiothoracic wound model, we compared the degree of air displacement achieved by a new insufflation device, a gas-diffuser, with that of a thin open-ended tube during steady-state and with carbon dioxide flows of 2.5, 5, 7.5, and 10 L/min. We also studied air displacement at the start of and after discontinuation of carbon dioxide insufflation with the gas-diffuser and evaluated the influence of an open pleura. During steady state, the gas-diffuser produced efficient air displacement in the wound cavity model at carbon dioxide flows of > or = 5 L/min (< or = 0.65% remaining air), whereas the open-ended tube was inefficient (> or = 82% remaining air) at all studied carbon dioxide flows (P < 0.001). An open pleural cavity prolonged the time needed to obtain a high degree of air displacement in the wound cavity (P = 0.001). Carbon dioxide insufflation of the cardiothoracic wound cavity should be initiated at a carbon dioxide flow of 10 L/min at least 1 min before the incision of the heart and great vessels and should be continued at a carbon dioxide flow of at least 5 L/min until surgical closure. PMID- 12538173 TI - Strict thermoregulation attenuates myocardial injury during coronary artery bypass graft surgery as reflected by reduced levels of cardiac-specific troponin I. AB - We assessed the cardioprotective effects of perioperative maintenance of normothermia by determining the perioperative profile of troponin I, a highly cardiac-specific protein important in risk stratification of patients with acute ischemic events. Candidates for their primary coronary artery bypass grafting (CABG) were randomized into a new thermoregulation system group, Allon( thermoregulation (AT; n = 30), and a routine thermal care (RTC; n = 30) group. Anesthetic and operative techniques were similar in both groups. Intraoperative warming was applied before and after cardiopulmonary bypass (CPB) and up to 4 h after surgery. Perioperative temperature and hemodynamic data were recorded. Blood samples for creatine kinase (CK) and its isoform, MB (CK-MB), and for cardiac-specific troponin I (cTnI) were obtained at predetermined intervals throughout the entire operation. Core and skin temperatures were higher in the AT group at all time points. The systemic vascular resistance was lower and the cardiac index higher in the AT group at all intra- and postoperative time points. Increases in CK, CK-MB, and cTnI levels indicated intraoperative ischemic insult in all patients. The respective CK levels for the AT and RTC groups were 53.3 +/- 22.7 IU/L and 47.9 +/- 17.86 IU/L at the time of anesthesia and 64.7 +/- 45.6 IU/L and 47.8 +/- 19.4 IU/L 30 min after the onset of surgery, demonstrating thereafter a steep increase before the discontinuation of CPB. CK-MB mass concentrations in both groups behaved almost identically. Pre-CPB cTnI levels at anesthesia induction were 0.3 +/- 0 ng/mL in both groups, followed by a distinctive profile observed after separation from CPB: 28.1 +/- 11.4 ng/mL, 26.05 +/- 9.20 ng/mL, and 22.3 +/- 8.9 ng/mL at discontinuation from CPB, chest closure, and 2 h after surgery, respectively, in the RTC group, versus 0.6 +/- 4.6 ng/mL, 6.6 +/- 5.5 ng/mL, and 7.9 +/- 4.76 ng/mL at these three time points, respectively, in the AT group (P < 0.01 between groups at the specified time points). Contrary to conventional thinking about the benefits of hypothermia, maintenance of normothermia throughout the non-CPB phases during CABG was demonstrated to be important in attenuating myocardial ischemic injury. Insofar as troponin I was more sensitive than other tested markers, it may provide important data on possible protection from myocardial insult and on other cardioprotective measures. PMID- 12538174 TI - Bispectral index-guided anesthesia in patients undergoing aortocoronary bypass grafting. AB - In this prospective, randomized study, we compared hemodynamics, oxygenation, possible intraoperative awareness, and costs in 62 patients undergoing first-time elective coronary artery bypass grafting at 2 different levels of anesthesia. Depth of anesthesia was assessed with bispectral index (BIS). All patients were anesthetized with sufentanil/midazolam. The dosage of sufentanil/midazolam was adjusted to achieve a BIS level of 45-55 in 32 patients (Group BIS 50), whereas in 30 patients a BIS level of 35-45 was intended (Group BIS 40). Data were obtained at six different time points before, during, and after surgery. All patients were asked about possible intraoperative awareness on the third postoperative day. There were no significant differences of any hemodynamic or oxygenation variables at any time between the two groups. BIS 40 patients received significantly (P < 0.05) more sufentanil (BIS 40, 888 +/- 211 microg; BIS 50, 514 +/- 99 microg) and midazolam (BIS 40, 22.4 +/- 5.6 mg; BIS 50, 16.6 +/- 3.7 mg) than BIS 50 patients. The reduction in anesthetic drugs used saved euro;13.78/US$12.54 per patient (P < 0.05) in Group BIS 50, but one BIS electrode caused additional costs of 19.95 Euros/18.15 US dollars. Time to extubation was not significantly prolonged in Group BIS 40 (BIS 40, 14.3 +/- 4.6 h; BIS 50, 11.8 +/- 3.8 h). There was no explicit memory during anesthesia in either group. BIS guided reduction of anesthetic medication saved costs and did not increase the risk of intraoperative awareness. However, total costs were increased by monitoring BIS, because of the BIS electrodes. PMID- 12538175 TI - Patients with a history of type II heparin-induced thrombocytopenia with thrombosis requiring cardiac surgery with cardiopulmonary bypass: a prospective observational case series. AB - Heparin-induced thrombocytopenia with thrombosis (HITT) type II is a life threatening complication of heparin therapy that most often occurs after 5-10 days of exposure to heparin. Anticoagulation is a significant concern for patients with HITT type II being prepared for cardiac surgery requiring cardiopulmonary bypass (CPB). We report a case series of 12 patients with a history HITT type II who underwent CPB and cardiac surgery. Six patients did not express the antibody that mediates HITT type II immediately before surgery. Heparin was used as the anticoagulant for the duration of CPB only, and all these patients did well without thrombotic complications. Six patients expressed the antibody that mediates HITT type II immediately before surgery. Hirudin was used as the anticoagulant for CPB in these patients. The ecarin clotting time was used to guide hirudin therapy during CPB. The patients receiving hirudin did well, but they had a large amount of bleeding, required transfusions of multiple allogeneic blood products, and had a frequent rate of reexploration of the mediastinum after CPB. PMID- 12538176 TI - Blood-transfusion requirements and blood salvage in donors undergoing right hepatectomy for living related liver transplantation. AB - Living related liver donation for liver transplantation in adults including its risks is receiving increased attention. We present data from 44 liver donors focusing on transfusion requirements and avoidance of heterologous transfusion. The volume of blood transfused (both autologous from preoperative donation and heterologous) was assessed including that derived from intraoperative isovolemic hemodilution, cell-saver salvaged, and retransfused blood. Hemoglobin concentration and central venous pressure were measured at specified time points before and during surgery. Intraoperative blood loss was calculated and correlated to the duration of parenchymal transsection, liver volume resected, and central venous pressure. There were no specific anesthesia-evoked complications. In 4 donors, major bleeding (>2000 mL) occurred. Blood loss averaged 902 +/- 564 mL (SD), yielding a minimal mean hemoglobin concentration of 8.1 +/- 1.2 g/dL. One donor received 3 U of heterologous blood and 30 donors received autologous blood from their preoperative donation. An average of 592 +/- 112 mL of blood derived from perioperative acute isovolemic hemodilution was retransfused as was 421 +/- 333 mL of washed red cells from the cell-saving system. Avoidance of heterologous blood transfusion, application of blood-saving techniques, and efficient pain management are crucial for adult living liver donors. Transfusion of banked blood can be avoided in most patients when intraoperative cell salvage, preoperative autologous blood donation, and intraoperative hemodilution are combined. PMID- 12538177 TI - Isoflurane and desflurane uptake during liver resection and transplantation. AB - When reducing fresh gas flows, the course of the vaporizer dial settings required to maintain a constant end-expired concentration of a potent inhaled anesthetic becomes more dependent on the uptake pattern of the inhaled anesthetic. However, the uptake pattern of potent inhaled anesthetics during prolonged procedures remains poorly quantified. Therefore, we determined isoflurane and desflurane uptake (V(iso) and V(des), respectively) during liver resection (LR, n = 17) and orthotopic liver transplantation (OLT, n = 18) using a liquid injection closed circuit anesthesia technique maintaining the end-expired concentration at 0.8% and 4.5%, respectively. Individual and average uptake curves were fit to a series of mathematical functions and compared with the square root of time and four compartment models. Cumulative doses of isoflurane and desflurane after 1 and 3 h in the LR group and after 1, 3, and 8 h in the OLT group were correlated with demographic variables and each patient's average cardiac output and cardiac index. Average uptake was best described by a biexponential fit: V(iso) (LR) = 1.5 x (1 - e(-t x 0.525)) + 16.4 x (1 - e(-t x 0.00506)) (R(2) = 0.9996); V(iso) (OLT) = 1.4 + 3.1 x (1 - e(-t x 0.472)) + 26.7 x (1 - e(-t x 0.00307)) (R(2) = 0.9994); V(des) (LR) = 2.7 x (1 - e(-t x 0.763)) + 28.7 x (1 - e(-t x 0.00568)) (R(2) = 0.9984); and V(des) (OLT) = 1.4 x (1 - e(-t x 0.472)) + 26.7 x (1 - e(-t x 0.00307)) (R(2) = 0.9994). Uptake showed significant interindividual variability, and correlations between uptake variables and patient characteristics were inconsistent. The rate of uptake decreased more slowly then predicted by the uptake models. Because neither existing models nor patient characteristics accurately predict uptake in the individual patient, anesthesia techniques involving the use of low fresh gas flows will continue to have to rely on drug monitoring. However, the slowly decreasing rate of uptake during prolonged procedures suggests that the number of vaporizer adjustments to keep the end-expired concentration constant should be limited. PMID- 12538178 TI - The influence of crystalloid and colloid replacement solutions in acute normovolemic hemodilution: a preliminary survey of hemostatic markers. AB - Acute normovolemic hemodilution (ANH), in which blood for autologous use is collected immediately before the onset of surgical blood loss, is a recommended autologous blood procurement technique for blood conservation. Both crystalloid and colloid replacement fluids have been used to maintain normovolemia during ANH, but few data are available to justify the use of a particular replacement fluid. Therefore, we designed a prospective, randomized study to determine if the replacement fluid choice affects various coagulation variables and perioperative blood loss. Forty adult patients, ASA physical status 1-3, scheduled for ANH during radical prostatectomy were randomly assigned to one of four replacement fluid groups: (a) Ringer's lactate, (b) 5% albumin, (c) 6% dextran 70 (DEX), or (d) 6% hetastarch (HES). After the induction of a standardized general anesthetic, all patients underwent ANH to a final hemoglobin level of 9 g/dL. Complete blood count, prothrombin time, partial thromboplastin time, fibrinogen, factors V and VIII levels, bleeding time, and thromboelastography (TEG measurements were obtained at similar time points in the procedure. When compared with baseline, activated partial thromboplastin time decreased and factor VIII levels increased in the postanesthesia care unit in both the Ringer's lactate and 5% albumin groups. The DEX and HES groups demonstrated a decrease in TEG maximum amplitude between preoperative and postanesthesia care unit measurements and TEG alpha (angle) was decreased from baseline in the DEX group. The changes in factor VIII, activated partial thromboplastin time, and TEG measurements indicate that HES and DEX may attenuate the hypercoagulability related to surgery. PMID- 12538179 TI - Reducing unnecessary cross-matching: a patient-specific blood ordering system is more accurate in predicting who will receive a blood transfusion than the maximum blood ordering system. AB - Most blood transfusions are given in the operating room. Adoption of the Maximum Surgical Blood Ordering Schedule in the 1970s reduced the amount of blood unnecessarily cross-matched, but the national cross-match-to-transfusion ratio remains at approximately two-to-one. We tested the ability of a patient-specific blood ordering system (PSBOS) to more accurately predict potential operative transfusion. All adult patients who had blood cross-matched before surgery (February through June 1999) for elective operative procedures at the University of Michigan Hospital were identified. Complex surgeries were excluded. Surgeons estimated the expected blood loss for their surgeries, and the expected postoperative hematocrit was calculated using the patient's blood volume, the surgeon-defined expected blood loss, and preoperative hematocrit. Lowest tolerated hematocrit was set at 21% except in patients with coronary artery disease or who were ASA physical status III or more (28%). Sensitivity, specificity, positive predictive value, and negative predictive value of the PSBOS were calculated. Our analysis included 178 cases in which blood was cross matched before surgery, representing 69 different surgeries and 42 surgeons. Only 16% of patients received an intraoperative transfusion. Of the 156 patients that PSBOS predicted would not require an operating room transfusion, 139 were not transfused. Of the 21 patients PSBOS predicted would be transfused, 11 were. The sensitivity of the algorithm as tested was 41%, the specificity 93%, the positive predictive value was 55%, and the negative predictive value was 89%. We conclude that PSBOS, which includes patient and surgeon variables in transfusion prediction, is more accurate than the Maximum Surgical Blood Ordering Schedule, which uses only surgical procedure. IMPLICATIONS: Currently, many units of blood set aside for surgery are never required, resulting in extra work and expense for blood banks. A formula that included patient weight and hematocrit and typical surgery blood loss was used to predict who would require transfusions. We reduced the predicted number of patients who had blood set aside from 178 to 21. PMID- 12538180 TI - Intravascular volume replacement therapy with synthetic colloids: is there an influence on renal function? PMID- 12538181 TI - Bivalirudin monitored with the ecarin clotting time for anticoagulation during cardiopulmonary bypass. PMID- 12538182 TI - Traditional versus new needle retractable i.v. catheters in children: are they really safer, and whom are they protecting? AB - Retractable needle IV catheters are designed to reduce needle-stick injuries; their use is mandated by federal regulations. We undertook a prospective data collection with the "traditional" IV catheters (JELCO) versus the "new" (Angiocath Autoguard). Assignment of catheter type was randomized by week. Data collected included assessment of the difficulty of i.v. access; number of catheters used; and splatters or spills of blood on skin, linen, floor, clothing, and operating room table. There were 473 attempted insertions in 330 patients over 20 days. No needle-stick injuries occurred. Seventy-seven blood spills or splatters occurred in 42 patients. The number of splatters or spills was four times more with the new compared with the traditional catheters. There were significantly more total splatters or spills and patients who experienced splatters or spills with new catheters when they were placed by attendings but not when placed by trainees. Our study suggests that use of this technology by more experienced anesthesiologists may increase the risk of exposure of health care providers to blood-borne pathogens. Practitioners should choose the i.v. system that allows the most efficient venous access with the least potential for blood contamination. Hospitals should allow the choice to be made by the individuals using the devices. PMID- 12538183 TI - Midazolam premedication in children: a comparison of two oral dosage formulations on sedation score and plasma midazolam levels. AB - We compared two available oral formulations of midazolam with respect to sedation score and plasma midazolam levels in pediatric surgical patients 2-10 yr old. The commercially available oral syrup was compared with a mixture of the IV midazolam preparation in Syrpalta syrup at an equivalent concentration of 2 mg/mL. ASA status I-II patients were randomly assigned to receive 0.5 mg/kg of either the commercial syrup (Group 1) or the prepared mixture (Group 2) as anesthetic premedication. Observer's Assessment of Alertness/Sedation scores were obtained by a blinded observer at 15 and 30 min after drug administration. Plasma midazolam levels were acquired exactly 45, 60, and 90 min after administration. Group 2 patients had a significantly lower median Observer's Assessment of Alertness/Sedation score (Group 1, 17; Group 2, 15) at 30 min (P < 0.03) and significantly higher mean plasma midazolam levels at all three acquisition times (mean +/- SD) (45 min: 63.1 +/- 23.9 ng/mL, Group 2; 43.4 +/- 19.6 ng/mL, Group 1; 60 min: 45.8 +/- 18.2 ng/mL, Group 2; 30.8 +/- 17.9 ng/mL, Group 1; 90 min: 28.9 +/- 12.6 ng/mL, Group 2; 21.0 +/- 8.9 ng/mL, Group 1) (P < 0.02). We conclude that IV midazolam mixed in Syrpalta syrup yields more reliable sedation and correspondingly higher plasma levels than an equivalent dose of the commercially formulated and marketed preparation. PMID- 12538185 TI - Anesthetic uptake of sevoflurane and nitrous oxide during an inhaled induction in children. AB - The uptake of sevoflurane and nitrous oxide (N(2)O) was characterized during the mask induction of anesthesia in healthy children. We assessed concentration and second gas effects by determining the influence of two different inspiratory N(2)O concentrations on the rate at which the estimated alveolar concentration (FA) increased to the inspired gas concentration (FI). Eighteen children aged 4 12 yr old were randomly assigned to receive a 6% sevoflurane mixture with either a large or a small N(2)O concentration with balance O(2). End-tidal and inspiratory concentrations of respiratory and anesthetic gases were continuously assessed during the induction. The FA/FI for the small N(2)O was 0.87 +/- 0.09 (mean +/- SD) and increased to 0.92 +/- 0.08 for the large N(2)O (P < 0.01). Both groups differed significantly at 3, 4, and 5 min. The FA/FI for sevoflurane increased but more slowly than for N(2)O. The mean only differed significantly at 3 min. Equilibration between FA and FI for N(2)O and sevoflurane was attained rapidly. Consistent with their respective blood/gas partition coefficients, the FA/FI for N(2)O increased more rapidly than that for sevoflurane. Increasing FI N(2)O produced a leftward shift in gas equilibration curves. A concentration effect was confirmed with N(2)O and a brief second gas effect, probably explained by the higher solubility of sevoflurane. PMID- 12538184 TI - EMLA cream versus dorsal penile nerve block for postcircumcision analgesia in children. AB - We conducted a prospective, randomized, double-blinded trial comparing preoperative application of EMLA cream and sodium chloride solution dorsal penile block (n = 31) with placebo cream and bupivacaine dorsal penile nerve block (n = 32) for postcircumcision analgesia. Pain was assessed using modified Children's Hospital of Eastern Ontario Pain Scale and the duration of block by the time to requirement of first dose of postoperative analgesic. There was no difference in Children's Hospital of Eastern Ontario Pain Scale between the two groups, but bupivacaine dorsal penile nerve block resulted in longer analgesia (P = 0.003). There were no local or systemic complications related to either technique, and there was a very small incidence of vomiting. We conclude that preoperative application of EMLA cream is an effective and simple method to produce postcircumcision analgesia with a very small incidence of adverse effects. PMID- 12538186 TI - Salicylate action on medullary inspiratory neuron activity in a brainstem-spinal cord preparation from newborn rats. AB - Salicylate affects central respiratory control. The inspiratory neurons are the most important component of the medullary respiratory control center because they modulate the final motor output via the phrenic nerve. We investigated changes in burst rate, intraburst firing frequency, and membrane properties of inspiratory neurons in the isolated brainstem after the administration of salicylate. Newborn rat brainstem-spinal cord preparations were superfused with salicylate. Whole cell recordings were performed from inspiratory neurons. Application of 1 mM salicylate caused an increase in the inspiratory neuronal burst rate from 6.9 +/- 1.6 bursts/min to 8.2 +/- 1.9 bursts/min (P < 0.05). The inspiratory neuron burst rate decreased from 8.3 +/- 0.7 bursts/min to 4.5 +/- 1.1 bursts/min after the application of 10 mM salicylate (P < 0.01). The depressant effect of 10 mM salicylate was antagonized by the gamma-aminobutyric acid (GABA) receptor antagonist bicuculline (1 microM). Resting membrane potential and intraburst firing frequency did not change with the application of salicylate and bicuculline even when the burst rate did change. We conclude that the effects of salicylate on the medullary inspiratory neurons are mainly due to a presynaptic action. GABAergic mechanisms are probably involved in the salicylate-induced central respiratory depression. PMID- 12538187 TI - Awake caudal anesthesia for inguinal surgery in one conjoined twin. PMID- 12538188 TI - Patient-controlled perineural analgesia after ambulatory orthopedic surgery: a comparison of electronic versus elastomeric pumps. PMID- 12538189 TI - The impact of economics on changing medical technology with reference to critical care medicine in the United States. PMID- 12538190 TI - Pulse oximetry for perioperative monitoring: systematic review of randomized, controlled trials. AB - Monitoring with pulse oximetry might improve patient outcome by enabling an early diagnosis and, consequently, correction of perioperative events that might otherwise cause postoperative complications or even death. The aim of the study was to clarify the effect of perioperative monitoring with pulse oximetry and to identify the adverse outcomes that might be prevented or improved by its use. Trials were identified by computerized searches of the Cochrane Library, MEDLINE, EMBASE, and by checking the reference lists of trials and review articles. All controlled trials that randomized patients to either pulse oximetry or no pulse oximetry during the perioperative period, including in the operating and recovery room, were included in the study. The search identified six reports. Of these 6 reports, 4 studies with data from 21,773 patients were considered eligible for analysis. Two studies specifically addressed the outcomes in question; both found no effect on the rate of postoperative complications using perioperative pulse oximetry. Hypoxemia was reduced in the pulse oximetry group both in the operating room and in the recovery room. During observation in the recovery room, the incidence of hypoxemia in the pulse oximetry group was 1.5-3 times less. There were postoperative complications in 10% of the patients in the oximetry group and in 9.4% in the control group. The duration of hospital stay was a median of 5 days in both groups, and an equal number of in-hospital deaths was registered in both groups. The studies confirmed that pulse oximetry could detect hypoxemia and related events. However, given the relatively small number of patients studied and the rare events being sought, the studies were not able to show an improvement in various outcomes. PMID- 12538191 TI - Endotracheal intubation device with a charge couple device camera. AB - We developed an orotracheal intubation device equipped with a charge couple device (CCD) camera, providing a wide field of vision. We used this device to perform endotracheal intubations in 62 anesthetized patients undergoing dental treatment and oral surgery. The time required to perform an endotracheal intubation with this system was examined. The use of this system is described below. The wand with the CCD camera was inserted into the oropharyngeal cavity. The oropharynx, including the epiglottis and glottis, could be visualized on the monitor screen. The tube introducer was inserted into the trachea through the vocal cords via the side tube of the wand. The wand with the CCD camera was withdrawn, leaving the tube introducer in the trachea. The endotracheal tube was then inserted into the trachea by using the tube introducer as a guide. The time required for the procedure was determined. The mean total time for the procedure was 41.2 s (maximum, 155 s; minimum, 14 s). There were no significant differences in this procedure when the patients were grouped according to the Cormack and Lehane classification. There was no failure to intubate using this system. Because the device can extensively visualize not only the larynx, glottis, and vocal cords, but also the movement of the tube introducer, on the monitor screen via the CCD camera, endotracheal intubation can be easily performed while the vocal cords are visualized on the monitor screen. PMID- 12538192 TI - The bispectral index in the diagnosis of perioperative stroke: a case report and discussion. PMID- 12538193 TI - The duration of immobilization causes the changing pharmacodynamics of mivacurium and rocuronium in rabbits. AB - In the clinical setting, in patients with a cast, it is not known whether the monitoring of the neuromuscular paralysis induced by either mivacurium or rocuronium in the contralateral limb is the correct interpretation. We compared the dose-response relationships and the neuromuscular blocking effects of mivacurium and rocuronium in 56 anesthetized rabbits immobilized in a plaster cast for 2, 4, and 6 wk. Train-of-four stimuli were simultaneously applied every 10 s to both common peroneal nerves, and the force of contraction of both tibialis anterior muscles was measured. Immobilization was associated with a rightward shift of the mivacurium and rocuronium dose-response curves after the duration of the immobilized limb, whereas no shift occurred in the contralateral limb. The 50% effective dose values for 0, 2, 4, and 6 wk of immobilization in the immobilized limb of mivacurium were 15.1 +/- 1.4, 18.2 +/- 1.5, 21.5 +/- 1.9, and 27.8 +/- 2.5 microg/kg, respectively, and they were unchanged in the contralateral limb. The calculated 50% effective dose values for the correspondence of rocuronium were 48.1 +/- 4.1, 56.2 +/- 4.2, 64.8 +/- 4.9, and 75.1 +/- 5.5 microg/kg, respectively, and they were unchanged in the contralateral limb. The rabbits receiving mivacurium and rocuronium had a significantly accelerated recovery from neuromuscular blockade compared with the placebo group in the immobilized limb after the immobilized duration, whereas there were no differences in the contralateral limb. The results of the present study showed that immobilization disuse atrophy produced by casting led to the development of resistance to both mivacurium and rocuronium; however, no resistance was shown in the contralateral limb. The peripheral nerve stimulator could be applied on the nonimmobilized limb, which might be associated with a normal recording if either mivacurium or rocuronium was used as neuromuscular relaxants. PMID- 12538194 TI - Nitrous oxide and xenon inhibit the human (alpha 7)5 nicotinic acetylcholine receptor expressed in Xenopus oocyte. AB - The neuronal nicotinic acetylcholine (nACh) receptor is one of the ligand-gated ion channels that regulate the synaptic release of neurotransmitters in the central nervous system. Recently, neuronal nACh receptors have received attention as a potential target for general anesthetics because many general anesthetics inhibit their functions at clinical concentrations. Several general anesthetics are known to inhibit the homomeric (alpha(7))(5) nACh receptor, a subtype of neuronal nACh receptors, but the effects of two gaseous anesthetics, nitrous oxide (N(2)O) and xenon (Xe), remain unknown. Using the two-electrode voltage clamping technique, we investigated the effects of N(2)O and Xe at the human (alpha(7))(5) nACh receptor expressed in Xenopus oocytes. At clinically relevant concentrations, N(2)O and Xe reversibly inhibited the ACh-induced currents of the (alpha(7))(5) nACh receptor in a concentration-dependent manner. The inhibitory actions of both anesthetics at the (alpha(7))(5) nACh receptor were noncompetitive and voltage-independent. Our results suggest that inhibition of the (alpha(7))(5) nACh receptor by N(2)O and Xe may play a role in their anesthetic effects. PMID- 12538195 TI - Propofol impairs the central but not the peripheral part of the motor system. AB - Propofol provides some degree of muscle relaxation. Previous studies have investigated the effects of propofol on either the central or peripheral parts of the motor system. In this study, we simultaneously assessed both central (spinal) and peripheral effects. In 15 patients, general anesthesia was induced and maintained with fentanyl and midazolam. Neuromuscular blocking drugs were not administered. To investigate the central portion of the motor system, we monitored spinal F waves, an electrophysiologic variable of alpha-motoneuron excitability. Direct electrophysiologic muscle responses (M waves) and mechanomyography were studied to detect the peripheral effects of propofol on neuromuscular transmission or muscle contraction strength. After baseline recordings, 3 IV boluses of propofol (2 times 1 mg/kg followed by 2 mg/kg) were administered at 5-min intervals. Mean F-wave amplitudes were significantly reduced compared with baseline measurements (mean +/- SD, 0.22 +/- 0.13 mV) after the first (0.13 +/- 0.08 mV; P < 0.05), second (0.08 +/- 0.09 mV; P < 0.05), and third (0.03 +/- 0.04 mV; P < 0.01) propofol injections. M-wave amplitudes and mechanomyography signals remained unchanged. Our data suggest that the central part, but not the peripheral part, of the motor system is impaired after bolus administration of propofol. PMID- 12538196 TI - The dose-dependent effects of fentanyl on rat skeletal muscle microcirculation in vivo. AB - Determining the effects of analgesia on the microcirculation is difficult because the surgery needed to allow in vivo observation often requires anesthesia. In this study, we used the dorsal microcirculatory chamber (DMC) to determine the effects of large (LF) and small (SF) dose IV fentanyl on the microcirculation compared with a conscious control. Male Wistar rats (130 g, n = 5) were implanted with the DMC to enclose a single layer of striated muscle. Animals were allowed 3 wk to recover from surgery and then, over the following 2 wk (1 infusion/wk) using intravital microscopy, the microcirculation was viewed in conscious animals (t = 0-30 min), followed by an induction bolus dose (t = 40-45 min), then a "step up" maintenance infusion of one of the following, LF (40-90 microg x kg(-1) x h( 1)), SF (10-60 microg x kg(-1) x h(-1)), or saline (5-10 microg x kg(-1) x h(-1)) (t = 45-105 min). Small arterioles (<30 micro m) dilated (23.6% +/- 7.1%) after induction with LF, but constricted (-21.3% +/- 7.1%) with SF (P < 0.05). During maintenance, constriction increased with increasing dose of LF (-21.9% +/- 4.0%) and SF (-16.7% +/- 9.1%) (t = 105 min, P < 0.05). Similar patterns were observed in all arterioles (10-120 microm) and venules (15-250 microm). We conclude that the DMC provides an excellent technique for observing microcirculatory responses to fentanyl, and in rat skeletal muscle in vivo, an i.v. infusion of fentanyl produces significant constriction of arterioles. PMID- 12538197 TI - Epidural injections of indomethacin for postlaminectomy syndrome: a preliminary report. AB - Since there have been side effects reported with the administration of corticosteroids epidurally, their application has been limited. Because some nonsteroidal antiinflammatory drugs have central and spinal antinociceptive actions, we have compared the effects of indomethacin (INM) given by the epidural route to methylprednisolone (MTP). This was a prospective, comparative study in an ambulatory pain care center. Two hundred six patients with recurrent low back pain (Visual Analog Scale >7) and radiculopathy after they had had 2 or more lumbar laminectomies with the diagnosis of "postlaminectomy syndrome" were randomly assigned to 1 of 3 groups. Group I (64 patients) was given 2 epidural injections of lyophilized INM 1 mg. Group II (60 patients) received 2 injections of 2 mg of INM at the same intervals. Group III (82 patients) was treated by 2 epidural injections of MTP 80 mg. In every case, the medication was diluted in 3 mL of 0.5% bupivacaine. Reductions of pain were assessed by changes in the Visual Analog Scale; physical activities, attitude, and medication intake were graded by the Pain Progress Score recorded before each treatment and 2 wk after the last. After each injection, all patients had pain relief to Visual Analog Scale <3. Increased analgesia (P < 0.05) was noted when a double dose of INM was used (Group II) or when 80 mg of MTP was given. The total average scores of the Pain Progress Score showed significant differences at the second injection in Groups II and III only. Physical activity, emotional attitudes, and medication intake were also improved but the changes were not statistically significant. In conclusion, in this group of patients, INM produced adequate analgesia in Groups I and II, with evidence suggesting that 2 mg of INM may produce a similar degree of pain relief as 80 mg of MTP after the second injection. Other nonsteroidal antiinflammatory drugs may be explored in the future for the same purpose. PMID- 12538198 TI - A comparison of multimodal perioperative analgesia to epidural pain management after gastric bypass surgery. AB - We compared pain intensity, analgesic consumption, patient satisfaction, and length of stay in 114 patients undergoing gastric bypass surgery under general anesthesia. Patients were randomized to incisional local anesthetic infiltration plus postoperative patient-controlled analgesia (Group A), epidural anesthesia and analgesia (Group B), or postoperative patient-controlled analgesia (Group C). All received perioperative nonsteroidal antiinflammatory drugs. Age, sex, body mass index, length of stay, and patient satisfaction were equivalent in all groups. Pain at time 0 and 36 h was the smallest in Group B, greater in Group A, and greatest in Group C. Pain scores in a subset of Group A were lower at all times than in Groups B and C, but this difference was significant only at 0, 12, and 36 h. In responders, infiltration analgesia as part of a multimodal regimen offers a simple, safe, and inexpensive alternative to epidural pain control. PMID- 12538199 TI - Postoperative pain relief after hepatic resection in cirrhotic patients: the efficacy of a single small dose of ketamine plus morphine epidurally. AB - In cirrhotic patients undergoing hepatic surgery, postoperative analgesia remains a challenge. In this study, we evaluated the efficacy of a single dose of morphine combined with small-dose ketamine given epidurally for postoperative pain relief. One-hundred-four classification "Child A" cirrhotic patients were randomly assigned to two groups: 1) (MKG, n = 54): epidural morphine (3.5-5 mg) plus ketamine (20/30 mg); and 2) epidural morphine (3.5/5 mg) (MG, n = 50). The level of analgesia, side effects, psychomimetic and neurological disorders, additional analgesic needs, and overall quality of the analgesia were recorded. The mean duration of analgesia was longer in the MKG group (27.2 +/- 8 h versus 16.4 +/- 10 h; P < 0.05). In the MKG group, the visual analog scale (VAS) score began to be significantly lower from 14 h at rest and 12 h on coughing until the end of the study. The need for additional analgesia was also smaller in the MKG group (P < 0.05): at 24 h, only 10% of patients in the MKG group needed complementary analgesia, whereas in the MG group it was 100% (P = 0.003). Side effects were similar in both groups. Psychomimetic side effects and neurological disorders were not detected. These results suggest that postoperative analgesia provided by a single dose of epidural morphine with small-dose ketamine is effective in cirrhotic Child's A patients having major upper abdominal surgery. PMID- 12538200 TI - Release of beta-endorphin immunoreactive material under perioperative conditions into blood or cerebrospinal fluid: significance for postoperative pain? AB - The function of beta-endorphin immunoreactive material (IRM) released under perioperative conditions remains to be clarified. In 17 patients undergoing orthopedic surgery, we determined beta-endorphin IRM in venous blood plasma and in cerebrospinal fluid (CSF) before surgery (t(A)); after termination of surgery and general anesthesia, but still under spinal anesthesia (t(B)); on occurrence of postoperative pain (t(C)); and 1 day after the operation (t(D)). Pain severity was rated by the patients by using a visual analog scale. Patients felt postoperative pain (t(C)), but they felt no pain at times t(A), t(B), and t(D). beta-Endorphin IRM plasma levels before surgery (t(A)) or with postoperative pain (t(C)) proved to be significantly higher than levels determined just after surgery, but still under spinal anesthesia (t(B)), or those determined 1 day after the operation (t(D)); beta-endorphin IRM plasma levels at times t(A) and t(C) correlated positively with postoperative pain severity (t(C)). beta Endorphin IRM CSF levels after surgery, but still under spinal anesthesia (t(B)), were significantly higher than levels determined at times t(A), t(C), or t(D). No correlation was found between beta-endorphin IRM CSF levels and pain severity. In conclusion, postoperative pain severity appears to be related to beta-endorphin IRM levels in plasma before surgery as well as with postoperative pain; the analgesic significance of this material remains to be elucidated. PMID- 12538201 TI - The significance of intrathecal catheter location in rats. AB - Although chronic intrathecal catheterization is a widely used method in rats, few calibration experiments have been performed. In this study, we investigated the correlation between the side position of the catheter tip and the side differences observed in the motor and sensory disturbances after intrathecal administration of lidocaine to a large number of rats. The existence of a sensory block was determined by the paw withdrawal test. The motor impairment was assessed by observing the complete clubbing of the hindpaw and measuring the hindpaw grip strength. After experimental use, we established the position of the catheter tip. The catheter tips were variously located in all directions of the transverse plane in the rat spinal subarachnoid space. Lidocaine administration (100 or 500 microg/5 microL; n = 264 and 112, respectively) led to dose-dependent motor and sensory disturbances. The effect of 100 microg of lidocaine exhibited side differences; i.e., the extents of both motor (r = 0.77) and sensory (r = 0.60 and r = 0.67 for the right and the left side, respectively) disturbances correlated significantly with the location of the catheter tip. Our data have shown that detection of the paralytic and/or antinociceptive effect of small-dose lidocaine before planned experiments is a simple and reliable method for prediction of the location of the catheter tip. We suggest that the position of the catheter might cause side differences in the drug effect, especially if small doses of drugs are administered and their effects are investigated on both sides. PMID- 12538204 TI - How to release allocated operating room time to increase efficiency: predicting which surgical service will have the most underutilized operating room time. AB - At many facilities, surgeons and patients choose the day of surgery, cases are not turned away, and staffing is adjusted to maximize operating room (OR) efficiency. If a surgical service has already filled its allocated OR time, but has an additional case to schedule, then OR efficiency is increased by scheduling the new case into the OR time of a different service with much underutilized OR time. The latter service is said to be "releasing" its allocated OR time. In this study, we analyzed 3 years of scheduling data from a medium-sized and a large surgical suite. Theoretically, the service that should have its OR time released is the service expected to have the most underutilized OR time on the day of surgery (i.e., any future cases that may be scheduled into that service's time also need to be factored in). However, we show that OR efficiency is only slightly less when the service whose time is released is the service that has the most allocated but unscheduled (i.e., unfilled) OR time at the moment the new case is scheduled. In contrast, compromising by releasing the OR time of a service other than the one with the most allocated but unscheduled OR time markedly reduces OR efficiency. OR managers can use these results when releasing allocated OR time. PMID- 12538202 TI - Minute sphere acupressure does not reduce postoperative pain or morphine consumption. AB - Minute sphere acupressure has been used for more than 2000 yr and remains popular in Japan. The points most relevant to abdominal surgery are those associated with meridian flows crossing or originating in the abdomen. We tested the hypothesis that minute sphere therapy reduces pain and analgesic requirements after open abdominal surgery. Participating patients were given standardized desflurane and fentanyl anesthetic. On completion of surgery, they were randomly assigned to untreated control or minute sphere acupressure at the Neiguan, Zusanli, Sanyinjiao, and Gongsun points. Each site was covered with bulky gauze dressings so that patients could not determine their assignments. Postoperative pain was treated with IV morphine via a patient-controlled analgesia pump. Our primary a priori end-points were pain and opioid consumption on the first postoperative morning. Data are reported as median (25th percentile, 75th percentile). Fifty three patients (30 controls and 23 minute spheres) completed the study. Morphine requirements (47 mg [27, 58] vs 41 mg [25, 69]) and pain scores (29.5 mm [16, 59] vs 40 mm [22, 58]) were similar in the control and acupressure groups. These data provide an 80% power for detecting a 50% difference in morphine consumption at an alpha of 0.05. Minute sphere treatment at the Neiguan, Zusanli, Sanyinjiao, and Gongsun points thus failed to provide analgesia after abdominal surgery. Minute sphere therapy is a form of acupuncture. We tested whether minute spheres placed on three acupressure points relevant to abdominal surgery reduced pain and morphine requirements after abdominal surgery. Treatment and control patients received a similar covering. Neither pain nor morphine requirements were different between the groups. PMID- 12538203 TI - The influence of protocol pain and risk on patients' willingness to consent for clinical studies: a randomized trial. AB - We tested the hypothesis that the risk or discomfort associated with a clinical trial influence patients' decisions to participate. Simultaneously, we evaluated factors likely to influence patients' decisions such as understanding of the risk and discomfort associated with the study, patient age, educational level, and psychological status. With IRB approval, participants, who believed they were being asked to participate in a real trial, were presented one of three sham protocols: no risk or pain (Control, n = 48), pain but no risk (Pain, n = 51), or risk but no pain (Risk, n = 51). Patients were debriefed at the end of the interview. Our major outcome measures were (a) understanding risk or pain associated with the proposed studies, (b) the extent to which patients felt pressured to participate, and (c) willingness to participate. Whereas understanding was similar in all groups (Control, 68%; Pain, 67%; and Risk, 72%), willingness to participate differed significantly (Control, 64%; Pain, 35%; Risk, 26%; P < 0.001). Patients who understood the level of risk or pain associated with the protocols were twice as likely to participate than those who did not (49% versus 24%; P = 0.003). Nine percent agreed to participate in the risky or painful protocols without understanding the risks involved. Patients who felt pressured did not agree to participate. Thus, the consent process protected patients, although for unexpected reasons. Understanding was poor, but patients who did not understand the risks or pain involved or who felt pressured rarely consented. Consequently, relatively few patients unknowingly agreed to participate in risky or painful studies. PMID- 12538205 TI - Declining proportion of publications by American authors in major anesthesiology journals. AB - A decline in the proportion of articles published by American authors in medical journals has been reported. We therefore sought to determine whether the contributions of authors from the United States to the three leading anesthesia journals changed between the years 1980 to 2000. The journals Pain, Anesthesiology, and Anesthesia & Analgesia were selected for evaluation on the basis of their respective impact factors. All clinical studies and basic science studies published in the years 1980, 1985, 1990, 1995, and 2000 were evaluated. The country of origin of the lead author of each article was determined by two of the investigators. chi(2) Tests and least squares linear regression analyses were used to determine associations between the source of publication (United States or abroad) and year of publication. The proportion of American publications in the leading anesthesia specialty journals was found to be decreasing over the period 1980-2000 because of an increase in the rate of publication from abroad that is disproportionate to the increase in the total number of publications in the journals over that time. The reasons for changes in anesthesia-related publications by American authors were not established by this study. The authors speculate that multiple factors are involved, including an increased emphasis on clinical care over research because of economic constraints, American publication in journals other than the leading specialty journals, and the increased quality of submissions from abroad. PMID- 12538206 TI - Adrenaline inhibits lipopolysaccharide-induced macrophage inflammatory protein-1 alpha in human monocytes: the role of beta-adrenergic receptors. AB - Macrophage inflammatory protein-1 alpha (MIP-1 alpha) has an important role in the development of inflammatory responses during infection by regulating leukocyte trafficking and function. Our study was conducted to investigate the effect of adrenaline on lipopolysaccharide (LPS)-induced MIP-1 alpha production by human peripheral blood monocytes and human monocytic THP-1 cells. Monocytes were incubated in vitro with LPS for 4 h at 37 degrees C in the presence and absence of adrenaline and/or specific alpha- and beta-adrenergic receptor antagonists and agonists. The effects of adrenaline on MIP-1 alpha synthesis were studied at the protein level by using enzyme-linked immunosorbent assays and at the messenger RNA level by using reverse transcriptase-polymerase chain reaction. Adrenaline inhibited LPS-induced MIP-1 alpha production in a dose-dependent manner. The suppressive effect could be completely prevented by propranolol, but not by phentolamine. The specific beta-adrenergic agonist isoproterenol produced the same inhibitory effect on LPS-induced MIP-1 alpha production, whereas the alpha-adrenergic agonist phenylephrine had a minimal effect. In addition, suppression of MIP-1 alpha production was associated with an increase of intracellular cyclic adenosine monophosphate (cAMP) by the cell membrane permeable cAMP analog dibutyryl-cAMP. Furthermore, we found that adrenaline inhibited LPS-induced MIP-1 alpha messenger RNA expression. These findings suggest that adrenaline can modulate MIP-1 alpha production in inflammatory diseases and sepsis. PMID- 12538207 TI - The effect of the alpha 2-agonist dexmedetomidine and the N-methyl-D-aspartate antagonist S(+)-ketamine on the expression of apoptosis-regulating proteins after incomplete cerebral ischemia and reperfusion in rats. AB - In this study, we investigated whether the neuroprotection previously seen with dexmedetomidine or S(+)-ketamine involves regulation of proapoptotic (Bax and p53) and antiapoptotic (Bcl-2 and Mdm-2) proteins. Rats were anesthetized with isoflurane. After surgical preparation of isoflurane was discontinued, animals were randomly assigned to receive fentanyl and nitrous oxide (N(2)O)/oxygen plus 100 microg/kg of dexmedetomidine intraperitoneally 30 min before ischemia (n = 8), 1 mg x kg(-1) x min(-1) of S(+)-ketamine and oxygen/air (n = 8), or fentanyl and N(2)O/oxygen (n = 8; control group). In all three treatment groups, incomplete cerebral ischemia (30 min) was induced by unilateral carotid artery occlusion and hemorrhagic hypotension to a mean arterial blood pressure of 30-35 mm Hg. Four hours after the start of reperfusion, the brains were removed, and the expression of apoptosis-regulating proteins was determined by using immunofluorescence and Western blot analysis. The results were compared with sham operated animals (n = 8). After cerebral ischemia/reperfusion, the relative protein concentration of Bax was increased by 110% in control animals compared with the dexmedetomidine- and S(+)-ketamine-treated rats and by 140% compared with the sham-operated animals. In animals treated with dexmedetomidine, the expression of Bcl-2 and Mdm-2 was larger compared with control (68% and 210%, respectively) or sham-operated (110% and 180%, respectively) animals. Therefore, it is possible that the neuroprotective properties of dexmedetomidine and S(+) ketamine seen in previous studies involve ultra-early modulation of the balance between pro- and antiapoptotic proteins. PMID- 12538208 TI - Potent sigma 1-receptor ligand 4-phenyl-1-(4-phenylbutyl) piperidine provides ischemic neuroprotection without altering dopamine accumulation in vivo in rats. AB - The in vivo signaling of ischemic neuroprotection provided by sigma-receptor ligands remains unclear. Catecholamines have been implicated in the propagation of ischemic neuronal injury, and previous in vitro studies suggest that sigma ligands modulate dopaminergic neurotransmission. In this study, we tested the hypothesis that the potent sigma(1)-receptor ligand 4-phenyl-1-(4-phenylbutyl) piperidine (PPBP) attenuates the increase of extracellular dopamine in ischemic striatum. Under controlled physiological conditions, a microdialysis probe was implanted in right caudoputamen (CP) complex of adult male Wistar rats. Rats were subjected to 2 h of transient middle cerebral artery occlusion (MCAO) by the intraluminal suture technique. In a blinded, randomized fashion, rats were divided into five treatment groups: Group 1 (n = 8; saline-saline) continuous i.v. infusion of saline vehicle 30 min before MCAO followed by saline at reperfusion until the end of the experiment; Group 2 (n = 8; PPBP-PPBP) i.v. PPBP 30 min before MCAO followed by 1 micromol x kg(-1) x h(-1) of PPBP; Group 3 (n = 8; saline-PPBP) i.v. saline before MCAO followed by PPBP; Group 4 (n = 4) surgical shams (saline-saline); and Group 5 (n = 4) surgical shams (PPBP-PPBP). Infarction volume at 22 h of reperfusion in the CP complex (percentage of ipsilateral structure) was significantly attenuated in rats treated with PPBP PPBP (27.3% +/- 9.1%) and saline-PPBP (27.8% +/- 12.7%) compared with saline saline (59.3% +/- 7.3%) treatment. There was a three- to fourfold increase in dopamine concentrations in the microdialysates within 40 min of the onset of MCAO. Dopamine and its metabolites dihydroxy phenylacetic acid and homovallinic acid levels were similar among the three groups subjected to MCAO. Therefore, PPBP provides significant ischemic neuroprotection in the CP complex without altering the acute accumulation of dopamine in vivo during transient focal ischemia in the rat. PMID- 12538209 TI - The vasodilatory effects of hydralazine, nicardipine, nitroglycerin, and fenoldopam in the human umbilical artery. AB - We studied the effects of hydralazine, nicardipine, nitroglycerin, and fenoldopam (a dopamine D1-agonist) on isolated human umbilical arteries (HUA) from patients classified as normotensive and with pregnancy-induced hypertension (PIH). Umbilical artery rings were contracted with the thromboxane A(2) analog (U46619; 10(-8) M) and then exposed to cumulative concentrations of fenoldopam, hydralazine, nicardipine, and nitroglycerin. Second, rings were preexposed to prazosin (10(-5) M), phenoxybenzamine (10(-5) M), or none, and the constriction responses to increasing doses of fenoldopam or dopamine were recorded. Nitroglycerin, hydralazine, and nicardipine produced concentration-dependent relaxation of U46619-preconstricted HUA segments from normotensive and PIH patients. Fenoldopam and dopamine induced umbilical artery constriction in both normal and PIH rings at concentrations > or = 10(-5) M and > or = 10(-3) M, respectively. Phenoxybenzamine, but not prazosin, pretreatment irreversibly abolished fenoldopam-induced contraction. In this in vitro study, nitroglycerin was the most potent vasodilator of the HUA constricted with U46619, followed by nicardipine and hydralazine. However, fenoldopam constricted HUA rings only at supratherapeutic concentrations. No significant differences of vascular responses to fenoldopam (P = 0.3534), nitroglycerin (P = 0.7416), nicardipine (P = 0.0615), and hydralazine (P = 0.5514) between rings from normotensive or hypertensive pregnant patients were shown. PMID- 12538210 TI - Spinal anesthesia for cesarean delivery shortly after an epidural blood patch. PMID- 12538211 TI - Perioperative epidural analgesia and outcome after major abdominal surgery in high-risk patients. AB - In a primary analysis of a large recently completed randomized trial in 915 high risk patients undergoing major abdominal surgery, we found no difference in outcome between patients receiving perioperative epidural analgesia and those receiving IV opioids, apart from the incidence of respiratory failure. Therefore, we performed a selected number of predetermined subgroup analyses to identify specific types of patients who may have derived benefit from epidural analgesia. We found no difference in outcome between epidural and control groups in subgroups at increased risk of respiratory or cardiac complications or undergoing aortic surgery, nor in a subgroup with failed epidural block (all P > 0.05). There was a small reduction in the duration of postoperative ventilation (geometric mean [SD]: control group, 0.3 [6.5] h, versus epidural group, 0.2 [4.8] h; P = 0.048). No differences were found in length of stay in intensive care or in the hospital. There was no relationship between frequency of use of epidural analgesia in routine practice outside the trial and benefit from epidural analgesia in the trial. We found no evidence that perioperative epidural analgesia significantly influences major morbidity or mortality after major abdominal surgery. PMID- 12538212 TI - Perioperative glucose infusion and the catabolic response to surgery: the effect of epidural block. AB - Although the nitrogen-sparing properties of epidural block and i.v. glucose on the days after surgical trauma have been well established, their metabolic effects during the acute phase of the stress response remain unclear. Therefore, in this study we investigated the effect of epidural block on glucose and protein kinetics during and immediately after surgery in patients receiving i.v. glucose at 2 mg x kg(-1) x min(-1). Sixteen patients undergoing colorectal surgery received either general anesthesia with epidural block with bupivacaine (EDA; n = 8) or general anesthesia alone (control; n = 8). Glucose and protein kinetics were determined during and 2 h after the operation by stable isotope tracers [6,6 (2)H(2)]glucose and L-[1-(13)C]leucine. Plasma concentrations of glucose, insulin, cortisol, and glucagon were also determined. Epidural block attenuated the perioperative increase in plasma glucose concentration (P < 0.05). The rate of appearance of glucose (R(a) glucose) and endogenous glucose production (EGP) were slower in the EDA group than in control subjects during (R(a) glucose, EDA 13.2 +/- 1.0 versus control 15.3 +/- 1.8 micromol x kg(-1) x min(-1); P < 0.05; EGP, EDA 1.2 +/- 1.2 versus control 3.8 +/- 1.7 micromol x kg(-1) x min(-1); P < 0.05) and after the operation (P > 0.05). Whereas protein breakdown and amino acid oxidation decreased in both groups (P < 0.05), whole-body protein synthesis remained unchanged. Insulin levels increased with both anesthetic techniques (P < 0.05). Intraoperative plasma concentrations of cortisol and glucagon were smaller in the EDA group (P < 0.05). The intraoperative suppression of EGP by exogenous glucose was more pronounced in the presence of epidural block. However, epidural block failed to exert a protein-sparing effect during the acute phase of the stress response in patients receiving i.v. glucose. PMID- 12538213 TI - Pharmacokinetics of ropivacaine in uremic and nonuremic patients after axillary brachial plexus block. AB - Reports on the efficacy and pharmacokinetics of local anesthetics in uremic patients have been controversial. Our study involved 29 uremic and 28 nonuremic patients. We performed axillary block with ropivacaine 300 mg (50 mL). Venous blood samples were drawn for 24 h for assay of total and unbound plasma ropivacaine, 3-hydroxyropivacaine, pipecoloxylidide (PPX), and serum alpha(1) acid glycoprotein (AAG). Block quality was similar in both groups. No toxicity occurred. Plasma clearance of ropivacaine was smaller and the area under the concentration-time curve of ropivacaine, 3-hydroxyropivacaine, and PPX larger in the uremic patients. The plasma concentration of PPX increased until 24 h in uremic patients whose AAG concentrations were also larger throughout the study. The free fraction of ropivacaine in plasma was smaller in the uremic group when measured 60 min and 12 h after the block, but the unbound concentration of ropivacaine was larger in the uremic group at 12 h. Enhanced absorption of ropivacaine into circulation, increased binding to AAG, and probably reduced urinary excretion of the metabolites lead to larger total plasma concentrations of ropivacaine and its main metabolites in uremic patients. PMID- 12538214 TI - Hypercarbic and hypoxic ventilatory responses after intrathecal administration of bupivacaine and sameridine. AB - Sameridine is a new compound with both local anesthetic and opioid properties (partial micro -opioid receptor agonist). It was intended for intrathecal administration to provide anesthesia for surgery and extended postoperative analgesia. In this double-blinded pharmacodynamic study with a two-parallel-group design, we investigated, during a 24-h period, the effects of intrathecal sameridine and bupivacaine on ventilation at rest and at ventilatory challenges during hypercarbia and hypoxia. Twenty-four healthy volunteers received either 25 mg of sameridine or 15 mg of bupivacaine intrathecally. Ventilation was measured by pneumotachography and in-line capnography. Sedation was rated by a visual analog scale. Segmental spread and development of motor and sensory block were similar in both groups. There was a decrease in tidal volume 2.5 to 6 h after injection in the bupivacaine group. This was seen only at 4 h in the sameridine group. There were no other major ventilatory differences between sameridine and bupivacaine during resting ventilation. Hypercarbic (tidal volume, mean inspiratory flow) and hypoxic (mean inspiratory flow) ventilatory responses were slightly decreased in the sameridine group, but not in the bupivacaine group. We conclude that intrathecal administration of sameridine or bupivacaine in healthy volunteers produces similar, minor effects on ventilatory responses over a 24-h observation period. PMID- 12538215 TI - The dose response and effects of dexamethasone on bupivacaine microcapsules for intercostal blockade (T9 to T11) in healthy volunteers. AB - Biodegradable microcapsules containing bupivacaine/dexamethasone produce an anesthetic duration of 7-11 days in animal models. In this investigation, we explored the effect of increasing doses (Part 1) and the effect of including dexamethasone (Part 2) on the onset, density, and duration of analgesia and anesthesia produced by bupivacaine microcapsules. Concentrations ranging from 0.3125% to 5.0% in microcapsules were compared with 0.25% aqueous bupivacaine (bilateral injection, three intercostal nerves, 2 mL per nerve) (Part 1). Part 2 compared 2.5% microcapsules with or without the inclusion of dexamethasone by unilateral blockade. Sensory block was assessed by pinprick, temperature sensation, and subjective numbness (0, not numb; 10, totally numb). Pharmacodynamic assessments and plasma drug concentrations of bupivacaine and dexamethasone were measured for 96 h. The onset time was reduced and the duration of analgesia increased over the 0.3125%-5.0% dose range (P < 0.02). Onset with 2.5% microcapsules approximated that of 0.25% aqueous bupivacaine. Microcapsule block duration increased to at least 96 h and was significantly longer than aqueous bupivacaine (P < 0.001). Inclusion of dexamethasone increased the duration of pinprick anesthesia in 2.5% microcapsules (P = 0.03). We conclude that bupivacaine/dexamethasone microcapsules are well tolerated and demonstrate a dose-related effect in onset and duration of intercostal blockade. Inclusion of dexamethasone increases intercostal block anesthesia. PMID- 12538216 TI - Prognostic significance of postoperative in-hospital complications in elderly patients. I. Long-term survival. AB - To determine the impact of in-hospital postoperative complications on long-term survival, we prospectively studied consecutive patients > or = 70 yr of age undergoing noncardiac surgery. Potential clinical risk factors were measured and evaluated for their association with the occurrence of long-term postoperative mortality. Long-term survival was determined by using the Kaplan-Meier method. Multivariate correlates of survival were analyzed with the Cox proportional hazards model. The survival of the study group was also compared with the age- and gender-matched general United States population. Five hundred seventeen patients who survived the initial hospitalization were studied. The mean follow up duration was 28.6 +/- 12.8 mo. One hundred sixty-four of 517 patients (31.7%) were deceased at the time of follow-up. A history of cancer (hazard ratio [HR] 2.44, 95% confidence interval [CI] 1.78-3.38, P < 0.0001), ASA physical status >II (HR 2.27, 95% CI 1.61-3.21, P < 0.0001), neurologic disease (HR 1.59, 95% CI 1.13-2.24, P = 0.008), age (HR 1.42 per decade, 95% CI 1.11-1.81, P = 0.005), postoperative pulmonary complications (HR 2.41, 95% CI 1.30-4.48, P = 0.005), and renal complications (HR 6.07, 95% CI 2.23-16.52, P < 0.0001) were significant independent predictors of decreased long-term survival. Compared with the United States population, patients with complications had a greater increase in mortality risk in the first 3 mo after surgery (HR 7.3 versus general population) than those without complications (HR 2.9, P = 0.023). An effort to improve perioperative care delivery to elderly surgical patients must include measures to minimize in-hospital postoperative complications, particularly those involving the pulmonary and renal systems. PMID- 12538217 TI - Prognostic significance of postoperative in-hospital complications in elderly patients. II. Long-term quality of life. AB - To determine the impact of nonfatal in-hospital postoperative complications on long-term quality of life and functional status, we studied consecutive patients > or = 70 yr of age who had undergone noncardiac surgery. The association between the occurrence of nonfatal in-hospital postoperative complications and long-term quality of life (measured by the Medical Outcome Study Short form 36) and functional status was determined 2-3 yr after surgery. Independent predictors of quality of life and functional status were measured by multivariate logistic regression. Two-hundred-sixty-four of 353 patients (74.8%) responded to the survey. The mean Medical Outcome Study Short form 36 scores of patients with in hospital postoperative complications were significantly lower than those without complications in the following domains: physical functioning (42.8 versus 53.9; P = 0.029), general health (52.3 versus 62.3; P = 0.02), and role emotional (45.7 versus 67.9; P = 0.00058). Patients who had postoperative complications were more likely to be dependent in daily living activities. Comparison with age-matched United States population showed that patients with postoperative complications had lower scores in physical functioning (42.8 versus 53.2; P = 0.04), role physical (26.6 versus 45.3; P = 0.0078), role emotional (45.7 versus 63.2; P = 0.025), and mental health (66 versus 74; P = 0.024). By multivariate logistic regression analysis, only a history of diabetes (odds ratios 4.2; 95% confidence interval, 1.7-10.3; P < 0.002 and new hospitalization because of medical reasons (odds ratio, 3.8; 95% confidence interval, 1.6-8.8; P < 0.002) were significant independent predictors of a long-term decrease in quality of life. Adjusting for other clinical factors, in-hospital complications no longer independently predicted changes in functional status. For geriatric patients, the occurrence of postoperative complications does not independently predict long-term quality of life or functional status. The important independent predictors are co-morbid conditions, age, and new hospitalization after discharge. PMID- 12538218 TI - A comparison of the upper lip bite test (a simple new technique) with modified Mallampati classification in predicting difficulty in endotracheal intubation: a prospective blinded study. AB - We explored the possibility that a simple and single test could replace the modified Mallampati score for either a difficult or an unaccomplished tracheal intubation in an impending hypoxic patient. Three hundred adult patients were enrolled in this study. They were subjected to the following assessments: 1) oropharyngeal class according to the modified Mallampati criteria; 2) the new, upper lip bite criteria-class I = lower incisors can bite the upper lip above the vermilion line, class II = lower incisors can bite the upper lip below the vermilion line, and class III = lower incisors cannot bite the upper lip; and 3) laryngeal view grading according to Cormack's criteria. The incidence of difficult intubation was 5.7%. The upper lip bite test showed significantly higher specificity and accuracy than the modified Mallampati test (P < 0.001). Comparisons of sensitivity, positive and negative predictive values, between the two tests, however, did not reveal any significant differences (P > 0.05). In conclusion, the upper lip bite test is an acceptable option for predicting difficult intubation as a simple, single test. PMID- 12538219 TI - A comparison of the ProSeal laryngeal mask and the laryngeal tube in spontaneously breathing anesthetized patients. AB - In this multicenter, randomized study, we compared ease of insertion, postinsertion hemodynamic repercussion, quality of ventilation, and the capacity to achieve a "hands-free" anesthesia delivery between two new devices: the ProSeal laryngeal mask airway (PLMA) and the Laryngeal Tube (LT). The incidence of postoperative laryngopharyngeal discomfort was examined after short surgical interventions in spontaneously breathing patients. After induction with fentanyl and propofol, the respective airways were inserted into 70 adult ASA physical status I and II patients (35 patients in each group). First-attempt insertion success rates were more frequent for the PLMA (77% versus 51%; P < 0.05), but success rates were similar (100% versus 97%) after 3 attempts. The anesthesiologists considered that insertion of the PLMA was easier (P < 0.001). Expired tidal volume was larger with the PLMA (404.9 versus 328.4 mL; P < 0.005) and the ability to achieve hands-free ventilation was more frequent with the PLMA (32 versus 21 cases; P < 0.004). Positional maneuvers with the LT to correct ventilation deficiencies were not always completely effective (5 of 13). There were no differences in the incidence of intolerance, sore throat, dysphagia, and/or dysphonia between the two devices. We conclude that the PLMA showed greater ease of insertion and reliability than the LT for use in nonparalyzed anesthetized patients. PMID- 12538220 TI - The Brandt tube system attenuates the cuff deflationary phenomenon after anesthesia with nitrous oxide. AB - The Brandt tube system can limit excessive cuff pressure during nitrous oxide (N(2)O) anesthesia, but there is a lack of data assessing whether the Brandt tube system avoids cuff deflation after cessation of N(2)O administration. In this study, we recorded air-filled cuff pressures of the Mallinckrodt Brandt or Hi Contour (control) tracheal tubes (Mallinckrodt, Athlone, Ireland) during 67% N(2)O anesthesia and the cuffs were aspirated if the cuff pressure exceeded 22 mm Hg; 180 min later, O(2) was substituted for N(2)O. The cuff pressure of both groups significantly decreased after N(2)O anesthesia but the time required for the cuff pressure to return to the initial pressure was longer in the Brandt group than in the control group (76.5 +/- 35.2 min and 36.5 +/- 18.1 min, respectively; P = 0.03). The incidence of air leaks was more frequent in the control group than in the Brandt group (P = 0.015); changes in intracuff N(2)O were small in the Brandt group (6.6 +/- 1.2% to 3.4 +/- 0.9%) compared with those in the control group (46.2 +/- 3.8% to 18.6 +/- 5.6%). Therefore, the Brandt tube system attenuates the cuff deflationary phenomenon after N(2)O anesthesia, whereas repeated cuff deflation during N(2)O anesthesia causes cuff deflation after cessation of N(2)O, resulting in a possible risk of air leaks. PMID- 12538221 TI - Intraoperative colloid administration reduces postoperative nausea and vomiting and improves postoperative outcomes compared with crystalloid administration. AB - The debate over colloid versus crystalloid as the best solution for intraoperative fluid resuscitation is not resolved. Published studies have shown that mortality is not related to the specific fluid used for resuscitation. In addition, the quality of postoperative recovery between colloid and crystalloid has not been well investigated. In a prospective, blinded fashion, we investigated the effects of colloid and crystalloid resuscitation on nausea and vomiting and on the postoperative patient recovery profile. Patients undergoing major elective noncardiac surgery were randomized to receive 6% hetastarch in saline (HS-NS), 6% hetastarch in balanced salt (HS-BS), or lactated Ringer's solution (LR) on the basis of a fluid administration algorithm. The anesthetic was standardized. Hemodynamic targets included maintenance of arterial blood pressure, heart rate, and urine output within a predefined range. A postoperative morbidity survey was performed at baseline and daily after surgery. Ninety patients participated in the study, with 30 patients in each group. The amounts of study fluid (mean +/- SD) administered were 1301 +/- 1079 mL, 1448 +/- 759 mL, and 5946 +/- 1909 mL for the HS-NS, HS-BS, and LR groups, respectively (P < 0.05, HS-NS and HS-BS versus LR). Both the HS-NS and HS-BS (colloid) groups had a significantly less frequent incidence of nausea and vomiting, use of rescue antiemetics, severe pain, periorbital edema, and double vision. We concluded that intraoperative fluid resuscitation with colloid, when compared with crystalloid administration, is associated with an improvement in the quality of postoperative recovery. PMID- 12538222 TI - The Laryngeal Tube S: a modified simple airway device. PMID- 12538223 TI - The contributions of A. W. Hofmann. PMID- 12538224 TI - Epidural ketamine in healthy children--what's the point? PMID- 12538225 TI - Yet another bend in the wand! PMID- 12538226 TI - Agitation after desflurane and Dixon and Massey up-and-down method. PMID- 12538227 TI - Effects of systemic local anesthetics on perioperative ischemia reperfusion may be beneficial. PMID- 12538228 TI - A misleading conclusion and abstract. PMID- 12538229 TI - Combined spinal and epidural anesthesia for cesarean section in a patient with hypertrophic obstructive cardiomyopathy. PMID- 12538230 TI - A structural complication in the torque control blocker Univent: fracture of the blocker cap connector. PMID- 12538231 TI - Rapacuronium: premarket drug evaluation can be very effective for the identification of drug risks. PMID- 12538232 TI - Profound pain due to propofol injection triggered myocardial ischemia in a patient with a suspected pheochromocytoma. PMID- 12538233 TI - Kinking of the pilot tube prevents inflation of the cuff. PMID- 12538234 TI - Other stimuli add to effect of remifentanil on BIS. PMID- 12538235 TI - Computational and experimental analysis reveals a novel Src family kinase in the C. elegans genome. AB - MOTIVATION: The complete genomes of a number of organisms have already been sequenced. However, the vast majority of annotated genes are derived by gene prediction methods. It is important to not only validate the predicted coding regions but also to identify genes that may have been missed by these programs. METHODS: We searched the entire C.elegans genomic sequence database maintained by the Sanger Center using human c-Src sequence in a TBLASN search. We have confirmed one of the predicted regions by isolation of a cDNA and carried out a phylogenetic analysis of Src kinase family members in the worm, fly and several vertebrate species. RESULTS: Our analysis identified a novel tyrosine kinase in the C.elegans genome that contains functional features typical of the Src family kinases that we have designated as Src-1. The open reading frame contains a conserved N-terminal myristoylation site and a tyrosine residue within the C terminus that is crucial for regulating the activity of Src kinases. Our phylogenetic analysis of Src family members from C. elegans, Drosophila and other higher organisms revealed a relationship among Src kinases from C. elegans and Drosophila. PMID- 12538236 TI - Chemometric modelling based on 2D-fluorescence spectra without a calibration measurement. AB - MOTIVATION: 2D fluorescence spectra provide information from intracellular compounds. Fluorophores like trytophan, tyrosine and phenylalanin as well as NADH and flavins make the corresponding measurement systems very important for bioprocess supervision and control. The evaluation is usually based on chemometric modelling using for their calibration procedure off-line measurements of the desired process variables. Due to the data driven approach lots of off line measurements are required. Here a methodology is presented, which enables to perform a calibration procedure of chemometric models without any further measurement. RESULTS: The necessary information for the calibration procedure is provided by means of the a priori knowledge about the process, i.e. a mathematical model, whose model parameters are estimated during the calibration procedure, as well as the fact that the substrate should be consumed at the end of the process run. The new methodology for chemometric calibration is applied for a batch cultivation of aerobically grown S. cerevisiae on the glucose Schatzmann medium. As will be presented the chemometric models, which are determined by this method, can be used for prediction during new process runs. AVAILABILITY: The MATHLAB routine is free available on request from the authors. PMID- 12538237 TI - A study of accuracy and precision in oligonucleotide arrays: extracting more signal at large concentrations. AB - MOTIVATION: Despite the success and popularity of oligonucleotide arrays as a high-throughput technique for measuring mRNA expression levels, quantitative calibration studies have until now been limited. The main reason is that suitable data was not available. However, calibration data recently produced by Affymetrix now permits detailed studies of the intensity dependent sensitivity. Given a certain transcript concentration, it is of particular interest to know whether current analysis methods are capable of detecting differential expression ratios of 2 or higher. RESULTS: Using the calibration data, we demonstrate that while current techniques are capable of detecting changes in the low to mid concentration range, the situation is noticeably worse for high concentrations. In this regime, expression changes as large as 4 fold are severely biased, and changes of 2 are often undetectable. Such effects are mainly the consequence of the sequence specific binding properties of probes, and not the result of optical saturation in the fluorescence measurements. GeneChips are manufactured such that each transcript is probed by a set of sequences with a wide affinity range. We show that this property can be used to design a method capable of reducing the high intensity bias. The idea behind our methods is to transfer the weight of a measurement to a subset of probes with optimal linear response at a given concentration, which can be achieved using local embedding techniques. AVAILABILITY: Program source code will be sent electronically upon request. PMID- 12538238 TI - A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. AB - MOTIVATION: When running experiments that involve multiple high density oligonucleotide arrays, it is important to remove sources of variation between arrays of non-biological origin. Normalization is a process for reducing this variation. It is common to see non-linear relations between arrays and the standard normalization provided by Affymetrix does not perform well in these situations. RESULTS: We present three methods of performing normalization at the probe intensity level. These methods are called complete data methods because they make use of data from all arrays in an experiment to form the normalizing relation. These algorithms are compared to two methods that make use of a baseline array: a one number scaling based algorithm and a method that uses a non linear normalizing relation by comparing the variability and bias of an expression measure. Two publicly available datasets are used to carry out the comparisons. The simplest and quickest complete data method is found to perform favorably. AVAILABILITY: Software implementing all three of the complete data normalization methods is available as part of the R package Affy, which is a part of the Bioconductor project http://www.bioconductor.org. SUPPLEMENTARY INFORMATION: Additional figures may be found at http://www.stat.berkeley.edu/~bolstad/normalize/index.html PMID- 12538239 TI - Combinatorial image analysis of DNA microarray features. AB - MOTIVATION: DNA and protein microarrays have become an established leading-edge technology for large-scale analysis of gene and protein content and activity. Contact-printed microarrays has emerged as a relatively simple and cost effective method of choice but its reliability is especially susceptible to quality of pixel information obtained from digital scans of spotted features in the microarray image. RESULTS: We address the statistical computation requirements for optimizing data acquisition and processing of digital scans. We consider the use of median filters to reduce noise levels in images and top-hat filters to correct for trends in background values. We also consider, as alternative estimators of spot intensity, discs of fixed radius, proportions of histograms and k-means clustering, either with or without a square-root intensity transformation and background subtraction. We identify, using combinatoric procedures, optimal filter and estimator parameters, in achieving consistency among the replicates of a gene on each microarray. Our results, using test data from microarrays of HCMV, indicate that a highly effective approach for improving reliability and quality of microarray data is to apply a 21 by 21 top-hat filter, then estimate spot intensity as the mean of the largest 20% of pixel values in the target region, after a square-root transformation, and corrected for background, by subtracting the mean of the smallest 70% of pixel values. AVAILABILITY: Fortran90 subroutines implementing these methods are available from the authors, or at http://www.bioss.ac.uk/~chris. PMID- 12538240 TI - An associative analysis of gene expression array data. AB - MOTIVATION: We face the absence of optimized standards to guide normalization, comparative analysis, and interpretation of data sets. One aspect of this is that current methods of statistical analysis do not adequately utilize the information inherent in the large data sets generated in a microarray experiment and require a tradeoff between detection sensitivity and specificity. RESULTS: We present a multistep procedure for analysis of mRNA expression data obtained from cDNA array methods. To identify and classify differentially expressed genes, results from standard paired t-test of normalized data are compared with those from a novel method, denoted an associative analysis. This method associates experimental gene expressions presented as residuals in regression analysis against control averaged expressions to a common standard-the family of similarly computed residuals for low variability genes derived from control experiments. By associating changes in expression of a given gene to a large family of equally expressed genes of the control group, this method utilizes the large data sets inherent in microarray experiments to increase both specificity and sensitivity. The overall procedure is illustrated by tabulation of genes whose expression differs significantly between Snell dwarf mice (dw/dw) and their phenotypically normal littermates (dw/+, +/+). Of the 2,352 genes examined only 450-500 were expressed above the background levels observed in nonexpressed genes and of these 120 were established as differentially expressed in dwarf mice at a significance level that excludes appearance of false positive determinations. PMID- 12538241 TI - Rank order metrics for quantifying the association of sequence features with gene regulation. AB - MOTIVATION: Genome sequences and transcriptome analyses allow the correlation between gene regulation and DNA sequence features to be studied at the whole genome level. To quantify these correlations, metrics are needed that can be applied to any sequence feature, regardless of its statistical distribution. It is also desirable for the metric values to be determined objectively, that is, without the use of subjective threshold values. RESULTS: We compare two metrics for quantifying the correlation of DNA sequence features with gene regulation. Each of the metrics is calculated from a rank-ordering of genes based on the value of the sequence feature of interest. The first metric is the area under the curve for a receiver operator characteristic plot (ROC AUC), a common way of summarizing the tradeoff between sensitivity and specificity for different values of a prediction criterion. We call the second metric the mean normalized conditional probability (MNCP). The MNCP can be thought of as the predictive value of the sequence feature averaged over all regulated genes. The statistical significance (P-value) of each metric can be estimated from simulations. Importantly, the P-value of the MNCP metric is less dramatically affected by the presence of false positives among the set of co-regulated genes than is the ROC AUC. This is especially useful in analyzing gene sets identified by DNA microarray analysis because such data cannot distinguish direct regulation by transcription factor binding from indirect regulation. We demonstrate that these two metrics, taken together, are useful tools for defining the binding site representation and regulatory control regions that best explain the difference between genes that are regulated by a given transcription factor and those that are not. Applications to other gene features are also described. AVAILABILITY: A Python program for calculating the ROC AUC and MNCP metric values given input rank orders is available from ftp://ftp.bs.jhmi.edu/users/nclarke/MNCP/ PMID- 12538242 TI - Gene finding with a hidden Markov model of genome structure and evolution. AB - MOTIVATION: A growing number of genomes are sequenced. The differences in evolutionary pattern between functional regions can thus be observed genome-wide in a whole set of organisms. The diverse evolutionary pattern of different functional regions can be exploited in the process of genomic annotation. The modelling of evolution by the existing comparative gene finders leaves room for improvement. RESULTS: A probabilistic model of both genome structure and evolution is designed. This type of model is called an Evolutionary Hidden Markov Model (EHMM), being composed of an HMM and a set of region-specific evolutionary models based on a phylogenetic tree. All parameters can be estimated by maximum likelihood, including the phylogenetic tree. It can handle any number of aligned genomes, using their phylogenetic tree to model the evolutionary correlations. The time complexity of all algorithms used for handling the model are linear in alignment length and genome number. The model is applied to the problem of gene finding. The benefit of modelling sequence evolution is demonstrated both in a range of simulations and on a set of orthologous human/mouse gene pairs. AVAILABILITY: Free availability over the Internet on www server: http://www.birc.dk/Software/evogene. PMID- 12538243 TI - A generalized global alignment algorithm. AB - MOTIVATION: Homologous sequences are sometimes similar over some regions but different over other regions. Homologous sequences have a much lower global similarity if the different regions are much longer than the similar regions. RESULTS: We present a generalized global alignment algorithm for comparing sequences with intermittent similarities, an ordered list of similar regions separated by different regions. A generalized global alignment model is defined to handle sequences with intermittent similarities. A dynamic programming algorithm is designed to compute an optimal general alignment in time proportional to the product of sequence lengths and in space proportional to the sum of sequence lengths. The algorithm is implemented as a computer program named GAP3 (Global Alignment Program Version 3). The generalized global alignment model is validated by experimental results produced with GAP3 on both DNA and protein sequences. The GAP3 program extends the ability of standard global alignment programs to recognize homologous sequences of lower similarity. AVAILABILITY: The GAP3 program is freely available for academic use at http://bioinformatics.iastate.edu/aat/align/align.html. PMID- 12538244 TI - A naive Bayes model to predict coupling between seven transmembrane domain receptors and G-proteins. AB - MOTIVATION: An understanding of the coupling between a G-protein coupled receptor (GPCR) and a specific class of heterotrimeric GTP-binding proteins (G-proteins) is vital for further comprehending the function of the receptor within a cell. However, predicting G-protein coupling based on the amino acid sequence of a receptor has been a daunting task. While experimental data for G-protein coupling exist, published models that rely on sequence based prediction are few. In this study, we have developed a Naive Bayes model to successfully predict G-protein coupling specificity by training over 80 GPCRs with known coupling. Each intracellular domain of GPCRs was treated as a discrete random variable, conditionally independent of one another. In order to determine the conditional probability distributions of these variables, ClustalW-generated phylogenetic trees were used as an approximation for the clustering of the intracellular domain sequences. The sampling of an intracellular domain sequence was achieved by identifying the cluster containing the homologue with the highest sequence similarity. RESULTS: Out of 55 GPCRs validated, the model yielded a correct classification rate of 72%. Our model also predicted multiple G-protein coupling for most of the GPCRs in the validation set. The Bayesian approach in this work offers an alternative to the experimental approach in order to answer the biological problem of GPCR/G-protein coupling selectivity. AVAILABILITY: Academic users should send their request for the perl program for calculating likelihood probabilities at jack.cao@astrazeneca.com. SUPPLEMENTARY INFORMATION: The materials can be viewed at http://www.astrazeneca montreal.com/AZRDM_info/supporting_info.pdf. PMID- 12538245 TI - Knowledge acquisition, consistency checking and concurrency control for Gene Ontology (GO). AB - MOTIVATION: A critical element of the computational infrastructure required for functional genomics is a shared language for communicating biological data and knowledge. The Gene Ontology (GO; http://www.geneontology.org) provides a taxonomy of concepts and their attributes for annotating gene products. As GO increases in size, its ongoing construction and maintenance becomes more challenging. In this paper, we assess the applicability of a Knowledge Base Management System (KBMS), Protege-2000, to the maintenance and development of GO. RESULTS: We transferred GO to Protege-2000 in order to evaluate its suitability for GO. The graphical user interface supported browsing and editing of GO. Tools for consistency checking identified minor inconsistencies in GO and opportunities to reduce redundancy in its representation. The Protege Axiom Language proved useful for checking ontological consistency. The PROMPT tool allowed us to track changes to GO. Using Protege-2000, we tested our ability to make changes and extensions to GO to refine the semantics of attributes and classify more concepts. AVAILABILITY: Gene Ontology in Protege-2000 and the associated code are located at http://smi.stanford.edu/projects/helix/gokbms/. Protege-2000 is available from http://protege.stanford.edu. PMID- 12538246 TI - Positional candidate gene selection from livestock EST databases using Gene Ontology. AB - MOTIVATION: The number of expressed sequence tags (ESTs) in GenBank has now surpassed 200,000 for cattle and 100,000 for swine. The Institute of Genome Research (TIGR) has organized these sequences into approximately 60,000 non redundant consensus sequences (identified by TIGR Gene Indices) for cattle and 40,000 for swine. Anonymous ESTs are of limited value unless they are connected to function. Functional information is difficult to manage electronically because of heterogeneity of meaning and form among databases. The Gene Ontology (GO) Consortium has produced ontologies for gene function with consistent meaning and form across species. Linking livestock EST to gene function through similarity with sequences from other annotation-rich mammals could accelerate: (1) the discovery of positional candidate genes underlying a livestock quantitative trait locus (QTL) and (2) comparative mapping between livestock and other mammals (e.g. humans, mouse and rat). We initiated this investigation to determine if incorporation of the GO into the annotation process could accelerate livestock positional candidate gene discovery. RESULTS: We have associated livestock ESTs with GO nodes through sequence similarity to the NCBI Reference Sequences (RefSeq). Positional candidate genes are identified within minutes that otherwise required days. The schema described here accommodates queries that return GO nodes from terms familiar to biologists, such as gene name, alternate/alias symbol, and OMIM phenotype. AVAILABILITY: Scripts and schema are available on request from the authors. PMID- 12538247 TI - Fractional populations in multiple gene inheritance. AB - MOTIVATION: With complete knowledge of the human genome sequence, one of the most interesting tasks remaining is to understand the functions of individual genes and how they communicate. Using the information about genes (locus, allele, mutation rate, fitness, etc.), we attempt to explain population demographic data. This population evolution study could complement and enhance biologists' understanding about genes. RESULTS: We present a general approach to study population genetics in complex situations. In the present approach, multiple allele inheritance, multiple loci inheritance, natural selection and mutations are allowed simultaneously in order to consider a more realistic situation. A simulation program is presented so that readers can readily carry out studies with their own parameters. It is shown that the multiplicity of the loci greatly affects the demographic results of fractional population ratios. Furthermore, the study indicates that some high infant mortality rates due to congenital anomalies can be attributed to multiple loci inheritance. AVAILABILITY: The simulation program can be downloaded from http://won.hongik.ac.kr/~mhchung/index_files/yapop.htm. In order to run this program, one needs Visual Studio.NET platform, which can be downloaded from http://msdn.microsoft.com/netframework/downloads/default.asp. PMID- 12538248 TI - FluxAnalyzer: exploring structure, pathways, and flux distributions in metabolic networks on interactive flux maps. AB - MOTIVATION: The analysis of structure, pathways and flux distributions in metabolic networks has become an important approach for understanding the functionality of metabolic systems. The need of a user-friendly platform for stoichiometric modeling of metabolic networks in silico is evident. RESULTS: The FluxAnalyzer is a package for MATLAB and facilitates integrated pathway and flux analysis for metabolic networks within a graphical user interface. Arbitrary metabolic network models can be composed by instances of four types of network elements. The abstract network model is linked with network graphics leading to interactive flux maps which allow for user input and display of calculation results within a network visualization. Therein, a large and powerful collection of tools and algorithms can be applied interactively including metabolic flux analysis, flux optimization, detection of topological features and pathway analysis by elementary flux modes or extreme pathways. The FluxAnalyzer has been applied and tested for complex networks with more than 500,000 elementary modes. Some aspects of the combinatorial complexity of pathway analysis in metabolic networks are discussed. AVAILABILITY: Upon request from the corresponding author. Free for academic users (license agreement). Special contracts are available for industrial corporations. SUPPLEMENTARY INFORMATION: http://www.mpi magdeburg.mpg.de/projects/fluxanalyzer. PMID- 12538249 TI - Reconstruction of metabolic networks from genome data and analysis of their global structure for various organisms. AB - MOTIVATION: Information from fully sequenced genomes makes it possible to reconstruct strain-specific global metabolic network for structural and functional studies. These networks are often very large and complex. To properly understand and analyze the global properties of metabolic networks, methods for rationally representing and quantitatively analyzing their structure are needed. RESULTS: In this work, the metabolic networks of 80 fully sequenced organisms are in silico reconstructed from genome data and an extensively revised bioreaction database. The networks are represented as directed graphs and analyzed by using the 'breadth first searching algorithm to identify the shortest pathway (path length) between any pair of the metabolites. The average path length of the networks are then calculated and compared for all the organisms. Different from previous studies the connections through current metabolites and cofactors are deleted to make the path length analysis physiologically more meaningful. The distribution of the connection degree of these networks is shown to follow the power law, indicating that the overall structure of all the metabolic networks has the characteristics of a small world network. However, clear differences exist in the network structure of the three domains of organisms. Eukaryotes and archaea have a longer average path length than bacteria. AVAILABILITY: The reaction database in excel format and the programs in VBA (Visual Basic for Applications) are available upon request. SUPPLEMENTARY MATERIAL: Bioinformatics Online. PMID- 12538250 TI - A task framework for the web interface W2H. AB - SUMMARY: The W3H task framework allows the execution of compound jobs utilizing the description of work and data flows in a heterogeneous bioinformatics environment using meta-data information. By means of these descriptions, the task system can schedule the necessary execution of applications available in the environment, depending on rules specified in the meta-data. By integrating this task framework into the publicly available web interface W2H, similarly based on meta-data, web access and data management are immediately available for each task description. Authors of task descriptions can base their work on the underlying classes and objects to be able to describe dependency rules between previously independent applications. The result of a compound task is given as XML data that is translated according to XSLT data into web pages or plain text to report the result of the task to the user. AVAILABILITY: Within the HUSAR environment at DKFZ http://genome.dkfz-heidelberg.de/ PMID- 12538251 TI - QuickLIMS: facilitating the data management for DNA-microarray fabrication. AB - SUMMARY: QuickLIMS is a Microsoft Access-based laboratory information and management system, capable of processing all information for microarray production. The program's operational flow is protocol-based, dynamically adapting to changes of the process. It interacts with the laboratory robot and with other database systems over the network, and it represents a complete solution for the management of the entire manufacturing process. AVAILABILITY AND SUPPLEMENTARY INFORMATION: http://www.dkfz heidelberg.de/kompl_genome/Other/QuickLims/index.html PMID- 12538252 TI - An efficient algorithm for minimal primer set selection. AB - SUMMARY: We have developed U-PRIMER, a primer design program, to compute a minimal primer set (MPS) for any given set of DNA sequences. The U-PRIMER algorithm, which uses automatic variable fixing and automatic redundant constraint elimination to tackle the binary integer programming problem associated with the MPS selection problem. The program has been tested successfully with 32 adipocyte development-related genes and 9 TB-specific genes to obtain their respective MPSs. AVAILABILITY: A free copy of U-PRIMER implemented in C++ programming language is available from http://www.u-vision biotech.com PMID- 12538253 TI - Efficient selective screening of haplotype tag SNPs. AB - Haplotypes defined by common single nucleotide polymorphisms (SNPs) have important implications for mapping of disease genes and human traits. Often only a small subset of the SNPs is sufficient to capture the full haplotype information. Such subsets of markers are called haplotype tagging SNPs (htSNPs). Although htSNPs can be identified by eye, efficient computer algorithms and flexible interactive software tools are required for large datasets such as the human genome haplotype map. We describe a java-based program, SNPtagger, which screens for minimal sets of SNP markers to represent given haplotypes according to various user requirements. The program offers several options for inclusion/exclusion of specific markers and presents alternative panels for final selection. AVAILABILITY: The www-based program is available at http://www.well.ox.ac.uk/~xiayi/haplotype/index.html. PMID- 12538254 TI - Simulating haplotype blocks in the human genome. AB - SUMMARY: A bioinformatic tool was written to simulate haplotypes and SNPs under a modified coalescent with recombination. The most important feature of this program is that it allows for the specification of non-homogeneous recombination rates, which results in the formation of the so-called 'haplotype blocks' of the human genome. The program also implements different mutation models and flexible demographic histories. The samples generated can be very useful to better understand the architecture of the human genome and to investigate its impact in association studies searching for disease genes. AVAILABILITY: The SNPsim package is available at http://www.evolgenics.com/software PMID- 12538255 TI - Pedstrip: extracting a maximal subset of available, unrelated individuals from a pedigree. AB - Certain types of genetic analysis are simplified by assembling a collection of unrelated individuals, e.g. case-control experiments. If a family study is being performed then it will be necessary to extract subsets of unrelated, available individuals from pedigress. Our program provides an optimal method for performing this task. AVAILABILITY: The software is available, free of charge, on request from Oxagen Ltd. SUPPLEMENTARY INFORMATION: http://www.oxagen.co.uk PMID- 12538256 TI - AVA: visual analysis of gene expression microarray data. AB - SUMMARY: AVA (Array Visual Analyzer) is a Java program that provides a graphical environment for visualization and analysis of gene expression microarray data. Together with its interactive visualization tools and a variety of built-in data analysis and filtration methods, AVA effectively integrates microarray data normalization, quality assessment, and data mining into one application. AVAILABILITY: The software is freely available for academic users on request from the authors. PMID- 12538257 TI - Matrix2png: a utility for visualizing matrix data. AB - We describe a simple software tool, 'matrix2png', for creating color images of matrix data. Originally designed with the display of microarray data sets in mind, it is a general tool that can be used to make simple visualizations of matrices for use in figures, web pages, slide presentations and the like. It can also be used to generate images 'on the fly' in web applications. Both continuous valued and discrete-valued (categorical) data sets can be displayed. Many options are available to the user, including the colors used, the display of row and column labels, and scale bars. In this note we describe some of matrix2png's features and describe some places it has been useful in the authors' work. AVAILABILITY: A simple web interface is available, and Unix binaries are available from http://microarray.cpmc.columbia.edu/matrix2png. Source code is available on request. PMID- 12538258 TI - SEGID: identifying interesting segments in (multiple) sequence alignments. AB - SUMMARY: SEGID is a tool for finding conserved regions (regions of high scores) for a given (multiple) sequence alignment. It takes a (multiple) sequence alignment as its input and converts the alignment into a sequence of numbers, where each number is the alignment score of a column. Three algorithms are used to identify regions of high scores. A graphical interface is provided to present those identified regions. AVAILABILITY: Free from http://www.cs.cityu.edu.hk/~lwang/segid/subject to copyright restrictions. PMID- 12538259 TI - RnaViz 2: an improved representation of RNA secondary structure. AB - SUMMARY: RnaViz has been developed to easily create nice, publication quality drawings of RNA secondary structure. RnaViz 2 supports CT, DCSE, and RNAML input formats and improves on many aspects of the first version, notably portability and structure annotation. RnaViz is written using a hybrid programming approach combining pieces written in C and in the scripting language Tcl/Tk, making the program very portable and extensible. AVAILABILITY: Source code, binaries for Linux and MS Windows, and additional documentation are available athttp://rrna.uia.ac.be/rnaviz/ PMID- 12538260 TI - r8s: inferring absolute rates of molecular evolution and divergence times in the absence of a molecular clock. AB - SUMMARY: Estimating divergence times and rates of substitution from sequence data is plagued by the problem of rate variation between lineages. R8s version 1.5 is a program which uses parametric, nonparametric and semiparametric methods to relax the assumption of constant rates of evolution to obtain better estimates of rates and times. Unlike most programs for rate inference or phylogenetics, r8s permits users to convert results to absolute rates and ages by constraining one or more node times to be fixed, minimum or maximum ages (using fossil or other evidence). Version 1.5 uses truncated Newton nonlinear optimization code with bound constraints, offering superior performance over previous versions. AVAILABILITY: The linux executable, C source code, sample data sets and user manual are available free at http://ginger.ucdavis.edu/r8s. PMID- 12538261 TI - Chimera: construction of chimeric sequences for phylogenetic analysis. AB - SUMMARY: Chimera allows the construction of chimeric protein or nucleic acid sequence files by concatenating sequences from two or more sequence files in PHYLIP formats. It allows the user to interactively select genes and species from the input files. The concatenated result is stored to one single output file in PHYLIP or NEXUS formats. AVAILABILITY: The computer program, including supporting files and example files, is available from http://www.dalicon.com/chimera/. PMID- 12538262 TI - G-language Genome Analysis Environment: a workbench for nucleotide sequence data mining. AB - SUMMARY: G-language Genome Analysis Environment (G-language GAE) is an open source generic software package aimed for higher efficiency in bioinformatics analysis. G-language GAE has an interface as a set of Perl libraries for software development, and a graphical user interface for easy manipulation. Both Windows and Linux versions are available. AVAILABILITY: From http://www.g-language.org/ under GNU General Public License. CD-ROMs are distributed freely in major conferences. PMID- 12538263 TI - A profile hidden Markov model for signal peptides generated by HMMER. AB - SUMMARY: Although the HMMER package is widely used to produce profile hidden Markov models (profile HMMs) for protein domains, it has been difficult to create a profile HMM for signal peptides. Here we describe an approach for building a complex model of eukaryotic signal peptides by the standard HMMER package. Signal peptide prediction with this model gives a 95.6% sensitivity and 95.7% specificity. AVAILABILITY: The profile HMM for signal peptides, data sets, and the scripts for analyzing data are available for non-commercial use at http://share.gene.com/. PMID- 12538264 TI - MPID: MHC-Peptide Interaction Database for sequence-structure-function information on peptides binding to MHC molecules. AB - SUMMARY: Binding of short antigenic peptides to Major histocompatibility complex (MHC) proteins is the first step in T-cell mediated immune response. To understand the structural principles governing MHC-specific peptide recognition and binding, we have developed the MHC-Peptide Interaction Database (MPID), containing sequence-structure-function information. MPID (version 1.2) contains curated x-ray crystallographic data on 86 MHC peptide complexes, with precomputed interaction parameters (solvent accessibility, hydrogen bonds, gap volume and gap index). A user-friendly web interface and query tools will facilitate the development of predictive algorithms for MHC-peptide binding from a structural viewpoint. AVAILABILITY: Freely accessible from http://surya.bic.nus.edu.sg/mpid. PMID- 12538265 TI - TopDraw: a sketchpad for protein structure topology cartoons. AB - SUMMARY: Protein topology cartoons are a representation of structural data commonly used by structural biologists to illustrate the relationship between one dimensional sequence and three-dimensional structural data in a convenient two dimensional format. TopDraw is a simple, freely available TCL/Tk based drawing program designed specifically for the production of publication quality topology cartoons in a style commonly presented by structural biologists. AVAILABILITY: TopDraw is freely available under the terms of the GNU General Public License. It can be downloaded from http://stein.bioch.dundee.ac.uk/~charlie/scripts/topdraw.html. PMID- 12538267 TI - ARIA: automated NOE assignment and NMR structure calculation. AB - MOTIVATION: In the light of several ongoing structural genomics projects, faster and more reliable methods for structure calculation from NMR data are in great demand. The major bottleneck in the determination of solution NMR structures is the assignment of NOE peaks (nuclear Overhauser effect). Due to the high complexity of the assignment problem, most NOEs cannot be directly converted into unambiguous inter-proton distance restraints. RESULTS: We present version 1.2 of our program ARIA (Ambiguous Restraints for Iterative Assignment) for automated assignment of NOE data and NMR structure calculation. We summarize recent progress in correcting for spin diffusion with a relaxation matrix approach, representing non-bonded interactions in the force field and refining final structures in explicit solvent. We also discuss book-keeping, data exchange with spectra assignment programs and deposition of the analysed experimental data to the databases. AVAILABILITY: ARIA 1.2 is available from: http://www.pasteur.fr/recherche/unites/Binfs/aria/. SUPPLEMENTARY INFORMATION: XML DTDs (for chemical shifts and NOE crosspeaks), Python scripts for the conversion of various NMR data formats and the results of example calculations using data from the S. cerevisiae HRDC domain are available from: http://www.pasteur.fr/recherche/unites/Binfs/aria/ PMID- 12538266 TI - DPX: for the analysis of the protein core. AB - SUMMARY: In order to obtain an accurate description of the protein interior, we describe a simple and fast algorithm that measures the depth of each atom in a protein (dpx), defined as its distance (A) from the closest solvent accessible atom. The program reads a PDB file containing the atomic solvent accessibility in the B-factor field, and writes a file in the same format, where the B-factor field now contains the dpx value. Output structure files can be thus directly displayed with molecular graphics programs like RASMOL, MOLMOL, Swiss-PDB View and colored according to dpx values. AVAILABILITY: The algorithm is implemented in a standalone program written in C and its source is freely available at ftp.icgeb.trieste.it/pub/DPX or on request from the authors. PMID- 12538268 TI - Visualization and analysis of protein interactions. AB - SUMMARY: We have developed a new program called InterViewer for drawing large scale protein interaction networks in three-dimensional space. Unique features of InterViewer include (1) it is much faster than other recent implementations of drawing algorithms; (2) it can be used not only for visualizing protein interactions but also for analyzing them interactively; and (3) it provides an integrated framework for querying protein interaction databases and directly visualizes the query results. AVAILABILITY: http://wilab.inha.ac.kr/protein/ PMID- 12538269 TI - How should a supra-regional scientific organisation optimise the delivery of health care? PMID- 12538270 TI - Experimental coronary artery occlusion: relevance to off-pump cardiac surgery. AB - Mechanical coronary artery occlusion is required for minimally invasive direct coronary artery bypass and off-pump coronary artery bypass surgery. It is important that the method of occlusion be minimally traumatic. Chronic effects of these methods have never been studied. Temporary occlusion of coronaries utilizing suture snare, silastic loop snare, and bulldog clamp was carried out in 12 Yucatan pigs. Three animals each were sacrificed acutely and at 3, 6, and 12 months. The area of occlusion of each vessel was examined by light microscopy and the degree of damage recorded. In the animals sacrificed acutely, there was more damage using the suture snare than with the other 2 methods, but there was minimal damage at longer intervals. There was slight damage acutely and chronically with the bulldog technique. No damage was seen acutely with the silastic loop technique, but some late damage was found. The techniques of coronary artery dissection and occlusion used for minimally invasive and off-pump bypass surgery may contribute to early postoperative graft occlusion. PMID- 12538271 TI - Early surgical intervention for infective endocarditis. AB - Infective endocarditis remains a serious and complex disease with significant morbidity and mortality. Sixty cases of infective endocarditis were retrospectively reviewed, consisting of 41 males and 19 females aged 7 to 50 years (mean, 30 years). Congenital heart disease was diagnosed in 19 of the patients and rheumatic heart disease in 41. Congestive heart failure occurred in 36 and systemic embolism in 8 cases. Blood cultures were positive in only 21.7% of the cases, while vegetations were detected by 2-dimensional echocardiography in 70%. Elective surgery was performed in 57 patients and emergent operation for systemic arterial embolization and/or intractable congestive heart failure in 3 patients. Two patients required reoperation for postoperative bleeding. All but 2 patients had been followed up for 6 to 160 months with no evidence of reinfection. Three patients with mechanical valve implantation later died of intracranial bleeding due to over-anticoagulation. The remaining 55 resumed normal activity. The encouraging outcomes were the result of an aggressive diagnostic approach and early surgical intervention. PMID- 12538272 TI - Preoperative intraaortic balloon pump for salvage myocardial revascularization. AB - The benefit of preoperative intraaortic balloon pumping on hospital and 1-year mortality was assessed in 17 consecutive patients with cardiogenic shock who underwent salvage coronary artery bypass surgery between January 1996 and May 2000. Mean preoperative cardiac index was 1.57 +/- 0 23 L.min(-1).m(-2). Hemodynamic stability was achieved prior to surgery in 14 patients, and mean cardiac index increased to 2.28 +/- 0.64 L.min(-1).m(-2). Hospital mortality was 5.9% (1 patient). Five patients required postoperative intraaortic balloon pumping because of low cardiac output. The duration of intensive care, hospital stay, and the total procedural cost were similar to those reported for high-risk coronary bypass grafting. Cardiac event-free survival was 88.2% (15/17) at 1 year. Intraaortic balloon pumping promises to be an effective therapy for patients in cardiogenic shock who subsequently undergo surgical myocardial revascularization as a salvage procedure. PMID- 12538273 TI - Effect of oral anticoagulant during pregnancy with prosthetic heart valve. AB - This retrospective study aimed to evaluate the risks and outcome of oral anticoagulant use during pregnancy in women with prosthetic heart valves. Between December 1989 and November 1998, 192 females of childbearing age underwent heart valve replacement with a mechanical prosthesis. There were 37 pregnancies in 30 patients during follow-up. Pregnancy was terminated on medical grounds in 5 cases, there were 2 (6%) spontaneous abortions, and 1 (3%) premature birth of a normal baby who died 24 hours later due to asphyxia. The other 29 pregnancies (91%) went to full term and the mothers continued taking oral anticoagulants until a week before the expected date of delivery, then switched to heparin. There was no thromboembolism, valve thrombosis, or maternal mortality. Three babies (10%) had a skeletal deformity: nasal hypoplasia in all 3, with cleft pinna in 1. Continuation of oral anticoagulants during pregnancy provided adequate protection against thromboembolism and valve thrombosis, but the risks of fetal abnormalities and premature delivery should be explained to women of childbearing age with a mechanical valve prosthesis. PMID- 12538274 TI - Coarctation of the aorta in neonates and young infants: surgical experience. AB - A retrospective analysis of repair of aortic coarctation in young infants was conducted. Between April 1997 and December 2000, 21 patients under 4 months of age underwent repair of coarctation. Their mean age and weight were 41 42 days (range, 2 to 120 days) and 3.6 0.7 kg (range, 2.6 to 4.9 kg). The indications for surgery were congestive heart failure and/or shock. Diagnosis was made by 2 dimensional echocardiography with Doppler color flow imaging. Preoperative gradients ranged from 25 to 100 mm Hg. Aortic arch hypoplasia was present in 8 patients; 7 patients also had ventricular septal defect. Wide excision of the coarctation segment with extended end-to-end anastomosis was performed in 20 patients, while 1 required a Gore-Tex interposition graft between the left common carotid artery and the descending aorta. Subclavian angioplasty was performed to augment the anastomosis in 1 patient. There was no early mortality. One patient died 2 months after surgery. Follow-up examination revealed recoarctation in 5 patients (23.8%), all of whom underwent successful balloon dilatation. In conclusion, wider excision of the coarctation with extended end-to-end anastomosis reduces the chances of recoarctation. Percutaneous balloon angioplasty for treating recoarctation is effective in immediately reducing pressure gradients. PMID- 12538275 TI - Tricuspid valve detachment for transatrial closure of ventricular septal defects. AB - Tricuspid leaflet detachment improves visualization and accuracy of closure of ventricular septal defects via the transatrial route. Between July 1998 and March 2001, surgical correction was performed in 296 cases of isolated ventricular septal defect, 215 cases of tetralogy of Fallot, and 16 cases of double-outlet right ventricle. Of these, 132 patients (79 with isolated ventricular septal defect, 49 with tetralogy of Fallot, and 4 with double-outlet right ventricle) underwent transatrial repair with temporary detachment of tricuspid leaflets for ventricular septal defect closure. The septal leaflet was detached in most cases, with anterior or posterior leaflets being detached when indicated. Median duration of intensive care was 3.6 days, and median hospital stay was 7 days. There was no incidence of tricuspid regurgitation attributable to leaflet detachment, as confirmed by postoperative echocardiography. Reoperation was not required for a residual defect or tricuspid regurgitation. The benefits of temporary leaflet detachment for transatrial repair of various difficult defects far outweigh the risk of postoperative tricuspid regurgitation. PMID- 12538276 TI - Arch-first technique for aortic arch operation using branched graft. AB - To compare the arch-first technique with conventional aortic arch reconstruction 19 patients were randomly assigned to either procedure. Nine patients underwent the arch-first technique (group A) and 10 underwent the conventional technique (group B). There were no hospital deaths and no significant differences between groups in terms of intraoperative bleeding or the duration of operation cardiopulmonary bypass aortic crossclamping recovery from anesthesia or intensive care. The mean duration of retrograde cerebral perfusion via the superior vena cava was significantly shorter in group A (41.7 +/- 10.4 min) than group B (63.9 +/- 10 min). Transient neurologic dysfunction was noted in 4 (44%) patients in group A 6 (60%) in group B postoperatively but there was no permanent neurologic dysfunction in either group. The arch-first technique makes it possible to reduce the duration of cerebral ischemia retrograde cerebral perfusion via the superior vena cava reestablish antegrade cerebral perfusion earlier without damaging severely atheromatous arch vessels or conducting retrograde cerebral perfusion via a femoral artery. This technique has the potential to reduce the incidence of neurologic dysfunction. PMID- 12538277 TI - Comparison of off-pump versus conventional coronary revascularization. AB - Between May 1999 and August 2001, 231 patients underwent off-pump coronary bypass, of whom 171 required revascularization of the anterior vessels of the heart. They were compared with a group matched for age, sex, and risk factors undergoing revascularization of the same group of vessels under cardiopulmonary bypass. Mortality was comparable in both groups but the incidence of cerebrovascular accident, respiratory insufficiency, and renal failure was less in the off-pump group. Postoperative drainage and blood transfusion requirements were significantly less in the off-pump group. Intensive care and hospital stay were shorter in the off-pump patients. A considerable number of patients are potential candidates for off-pump coronary bypass, the only contraindication being technical limitations. Follow-up at 6 to 24 months indicates that off-pump coronary bypass can be performed safely with a decrease in morbidity and more rapid return to normal lifestyle. PMID- 12538278 TI - Combined valve operations with transection of ascending aorta. AB - To facilitate combined aortic and mitral valve operations, a new approach was developed. It was employed in 50 patients with both aortic and mitral valve disease. The ascending aorta was transected 1.5 cm above the aortic annulus, and the roof of the left atrium was incised. The damaged valves were replaced or repaired. Both aortic and mitral valves could be exposed satisfactorily by this approach. All patients recovered smoothly after the operation. Complications comprised intraoperative bleeding from the distal end of the ascending aorta in 1 patient and hoarseness in 3 postoperatively. The approach was considered quick and easy for combined valve operations. PMID- 12538279 TI - Infection risk factors in pediatric cardiac surgery. AB - Cardiac operations were preformed in 499 children from January 1998 through December 1999. Their median age was 263 days. A positive culture from blood, bronchoalveolar lavage, wound, or central catheter was obtained in 110 patients (22%). Age, sex, presence of pulmonary hypertension, body surface area, ratio of body surface area to oxygenator surface area, whether heart surgery was open or closed, and the duration of the operation, cardiopulmonary bypass, intubation, and intensive care were analyzed. Patients who developed infections were significantly younger, with smaller body surface areas and disparity with the oxygenator surface area, longer operative and bypass times, extended intubation, and prolonged intensive care. There was a significant correlation between infection and pulmonary hypertension. Sex and type of operation were not predictors of infection. PMID- 12538280 TI - Anomalous connection of left circumflex artery to pulmonary artery. AB - A 52-year-old woman was evaluated for chest discomfort and dyspnea on exertion. Coronary angiography demonstrated an anomalous connection between the left circumflex artery and the right pulmonary artery. Successful repair was achieved under cardiopulmonary bypass, and the patient became completely symptom-free. PMID- 12538281 TI - Ellis-van Creveld syndrome associated with thymic hypoplasia. AB - A 16-month-old girl presented with the classic signs of Ellis-van Creveld syndrome. Mitral clefts were repaired by direct suturing. The single atrium was repaired by creating a new septum with a pericardial patch, leaving the coronary sinus in the left atrium. During perioperative exploration, thymic hypoplasia was recognized. PMID- 12538282 TI - Noninvasive diagnosis of coronary artery fistula after cardiac surgery. AB - A 5-year-old boy underwent ventricular septal defect closure as well as resection of the muscle bundle of a double-chamber right ventricle and a subaortic membrane. Five days after surgery, echocardiography revealed a left anterior descending artery fistula with 2 sites of entry to the right ventricle. The child was asymptomatic with no murmurs. Echocardiography is an excellent tool for detecting such silent clinical lesions. PMID- 12538283 TI - Caught in the act: impending paradoxical embolism. AB - In a 73-year-old woman admitted with pulmonary embolism, a thrombus wedged across a patent foramen ovale was revealed by transesophageal echocardiography. After systemic anticoagulation, cardiac embolectomy and closure of the atrial septal defect were considered. Before surgery, repeat transesophageal echocardiography revealed only a small patent foramen ovale with no residual thrombus. We assumed dissolution of the thrombus as the patient had no sign of systemic or recurrent pulmonary embolism. PMID- 12538284 TI - Mitral annular reconstruction. AB - Mitral annular reconstruction using a pericardial patch was performed in 3 cases of atrioventricular disruption. This technique may be useful for atrioventricular disruption in cases of active endocarditis, redo valve replacement, left ventricular rupture after mitral valve replacement, and annular calcification. PMID- 12538285 TI - False aneurysm due to suture loosening after aortic arch replacement. AB - We describe a case of false aneurysm due to loosening of the anastomotic sutures 5 years after graft replacement for subacute type A aortic dissection. Leakage from the graft was controlled by tightening the suture slack indicating that suture loosening was the cause of the aneurysm. The mechanism was probably the progressive thinning of the edematous swollen aortic wall over the years causing the sutures to loosen. PMID- 12538286 TI - Acquired tracheoesophageal fistula following blunt trauma to the chest. AB - Tracheoesophageal fistula following blunt chest trauma is rare. Typically the patient is a young male with an elastic chest wall who is involved in a motor vehicle accident. In this case the victim was a motorcyclist who collided with a stationary lorry. He underwent surgery 4 weeks after the injury made an uncomplicated recovery. PMID- 12538287 TI - Bidirectional Glenn shunt for right ventricular endomyocardial fibrosis. AB - A 25-year-old man in New York Heart Association functional class IV with right ventricular endomyocardial fibrosis received a palliative bidirectional Glenn shunt. Despite a stormy postoperative convalescence the bidirectional Glenn shunt provided good long-term palliation. PMID- 12538288 TI - Acute descending aortic dissection associated with coarctation in adult. AB - A 44-year-old man with congenital aortic coarctation experienced acute dissection that crossed the coarctation and extended to both iliac arteries. The primary entry of the dissection was proximal to the coarctation segment, just below the origin of the left subclavian artery. Surgical procedures involved resection of the diseased segment including the coarctation, and graft interposition. PMID- 12538289 TI - Adult presentation of complete atrioventricular septal defect. AB - Complete atrioventricular septal defect generally presents early in life with symptoms of congestive heart failure. Untreated, patients almost universally develop irreversible pulmonary hypertension. We present an unusual case of a Down's syndrome patient whose septal defect first presented at 20 years of age. PMID- 12538290 TI - Severe unilateral pulmonary vascular changes in child with polysplenia. AB - A 12-year-old boy with polysplenia and single ventricle experienced recurrent episodes of pneumonia, hemoptysis, and pulmonary hypertension. Unilateral pulmonary vein obstruction was diagnosed, and a left pneumonectomy was performed. Microscopy of the resected specimen revealed pulmonary veno-occlusive disease in the small pulmonary venules, and old arteritis in the small pulmonary arteries. PMID- 12538291 TI - Cardiac operation with associated pulmonary resection: a word of caution. AB - Pulmonary tuberculosis reactivation is an unusual cause of respiratory failure after cardiac surgery. Fulminating tuberculosis was reactivated in a 50-year-old man after combined coronary artery bypass grafting and pulmonary resection on cardiopulmonary bypass. Clinicians should be aware of the immunosuppressive effects associated with cardiopulmonary bypass, and the consequent potential for tuberculosis reactivation. PMID- 12538292 TI - Neuroleptic malignant syndrome: uncommon postoperative diagnostic dilemma. AB - Neuroleptic malignant syndrome occurred in a 71-year-old man on haloperidol therapy for mild depressive dementia. After coronary artery bypass grafting, he developed hyperthermia, elevated creatine kinase without a corresponding rise in the MB-isoenzyme, leukocytosis, raised liver enzymes, urea, and creatinine. His condition responded to bromocriptine therapy. PMID- 12538293 TI - Acute right coronary artery embolus after aortic valve surgery. AB - A 35-year-old man with rheumatic aortic valve regurgitation developed acute right ventricular failure 8 days after valve replacement with a mechanical prosthesis. Angiography showed a right coronary artery embolus. Successful coronary artery bypass grafting was performed on the beating heart. PMID- 12538294 TI - Freestyle xenograft implantation technique for reducing aortic insufficiency. AB - The Freestyle aortic xenograft has been shown to be safe and effective for aortic valve replacement. However,implantation is often complicated by aortic valve insufficiency, which could lead to premature valve failure. We describe an implantation technique that can potentially prevent aortic insufficiency. PMID- 12538295 TI - Bronchoplasty with plication of the proximal bronchial membranous portion. AB - Bronchoplasty originally designed for patients with inadequate pulmonary function who cannot tolerate pneumonectomy, has recently yielded good results as well as offering an alternative to pneumonectomy. We describe a technique for plication of the proximal bronchial membranous portion when there is significant discrepancy in lumen size between the main and the segmental bronchi to allow precise end-to-end anastomosis. Plication helps prevent postoperative anastomotic stenosis and obstruction. PMID- 12538296 TI - Simplified elephant trunk graft technique for aortic arch replacement. AB - We refined the elephant trunk graft to facilitate and reinforce the distal anastomosis in aortic replacement operations. A cuff is created in a single four branch graft, which is used for the distal anastomosis; the trunk below the cuff is inserted into the distal aortic stump. This method is feasible for repairing extensive aortic aneurysm with a fragile wall and for treating acute aortic dissection where thromboocclusion of the remaining false lumen is desired. PMID- 12538297 TI - Heartport endoclamp for complex cardiac repairs with total circulatory arrest. AB - A method of performing redo cardiac operations using port-access technology and total circulatory arrest is described. The technique was useful in 2 cases requiring re-intervention within 4 months of the primary procedure. The indications were repair of an infected ventricular aneurysm and recurrence of a postinfarction ventricular septal defect. Dense mediastinal adhesions were avoided by approaching the site of pathology directly via a left anterior thoracotomy. PMID- 12538298 TI - Unexplained systemic hypertension after closure of ductus arteriosus. PMID- 12538299 TI - Unexplained systemic hypertension after closure of ductus arteriosus. PMID- 12538300 TI - Tonsillectomy. PMID- 12538301 TI - Caustic burn caused by potassium permanganate. PMID- 12538302 TI - Community child health training. PMID- 12538303 TI - Parenting programmes. PMID- 12538304 TI - The fat of the land. PMID- 12538305 TI - Classification of child abuse by motive and degree rather than type of injury. AB - The protection of children may be enhanced if ill treatment is classified by motive and degree rather than by type of injury. Four categories are proposed: A, abuse: premeditated ill treatment undertaken for gain by disturbed, dangerous, and manipulative individuals; B, active ill treatment: impulsively undertaken because of socioeconomic pressures, lack of education, resources, and support, or mental illnesses; C, universal mild ill treatment: behaviour undertaken by all normal caring parents in all societies; and D, neglect: defined here as an unintentional failure to supply the child's needs. Such a classification could clarify the procedures for investigation and protection, and support the creation of a Special Interagency Taskforce on Criminal Abuse (SITCA) for those suspected of abuse (category A). PMID- 12538306 TI - How to distinguish between neglect and deprivational abuse. AB - Neglect is a major cause of inadequate childcare in all societies and should be differentiated from abuse. "Neglect" is defined here, as the "neglectful" failure to supply the needs of the child, including emotional needs. It does not include the deliberate and malicious withholding of needs, which is a form of abuse. Neglect has its roots in ignorance of a child's needs and competing priorities; it is passive and usually sustained. The carer is without motive and unaware of the damage being caused. Malnutrition is a prime example of neglect; the stigma associated with the term abuse should never be applied to the poor struggling or uneducated mother whose child, that she loves dearly, becomes malnourished. Education of the mother and society and relief from the vicissitudes of poverty are required to alleviate most neglect of the world's children. PMID- 12538307 TI - Preventing sleeping problems in infants who are at risk of developing them. AB - AIMS: (1) To identify factors at 1 week of age which put infants at risk of failing to sleep through the night at 12 weeks of age. (2) To assess whether a behavioural programme increases the likelihood that these infants will sleep through the night at 12 weeks of age. METHODS: A community sample of 316 newborn infants was employed to identify the risk factors at 1 week of age which increased the likelihood of failing to sleep through the night at 12 weeks of age. Infants who met these risk criteria and were randomly assigned to a behavioural programme were compared with at risk infants in the control group on measures of sleeping, crying, and feeding at 12 weeks of age. RESULTS: Infants who had a high number (>11) of feeds in 24 hours at 1 week were 2.7 times (95% CI 1.5 to 4.8) more likely than other control group infants to fail to sleep through the night at 12 weeks of age. At 12 weeks, 82% of these at risk infants assigned to the behavioural programme, compared to 61% in the control group, slept through the night. The findings were similar in breast and bottle feeders. CONCLUSIONS: Preventing infant sleeping problems should be more cost effective than treating them after they have arisen. This study provides evidence that it is possible to identify infants who are at risk of failing to sleep through the night at an early age, and that a simple, three step, preventive behavioural programme increases the number who sleep through the night by 21%. PMID- 12538308 TI - Sudden infant death syndrome: bed sharing with mothers who smoke. AB - AIM: To examine the sleeping arrangements of sudden infant death syndrome (SIDS) cases on the Wirral. In particular to determine the prevalence of bed sharing with mothers who smoke, a known risk factor for SIDS. METHODS: Retrospective study of postmortem determined cases of SIDS from 1995 to 2000 on the Wirral peninsula (population 350 000, 3500 annual births). Ambulance crew reports, case notes, health visitor reports, postmortem reports, and case discussion records were studied for each case. RESULTS: There were 25 cases of SIDS in the postneonatal age group over the six year period. In nine cases the baby was bed sharing with the mother, of whom seven were smokers. Five of these mothers reported using alcohol or illicit drugs on the night of their infant's death. In two further cases the baby slept on a sofa with a parent. CONCLUSIONS: Bed sharing and smoking remain important risk factors for SIDS. Mothers should be advised ante- and postnatally of this combination of risk factors. Such advice should also include a recommendation not to sleep with their baby if under the influence of alcohol or illicit drugs, and never to sleep on a sofa with their baby. All "Child Health Record" books given to parents on the Wirral now include this advice. "Reduce the Risk" advice leaflets given to parents pre- and postnatally also now carry the recommendation, and health visitors and midwives have been educated with respect to these additions. PMID- 12538310 TI - Dumbbell tumour of the mediastinum. PMID- 12538309 TI - Disabilities and trends over time in a French county, 1980-91. AB - AIM: To describe trends over time and types of disability for children born in a French county from 1980 to 1991. METHODS: Data were collected from medical records of a morbidity register; disabled children with at least one severe deficiency have been included. Prevalence rates are given per 1000 resident children, over four three-year periods. RESULTS: Overall, 7.73 per 1000 children (that is, 1360 children), had a severe childhood disability, and the prevalence rate had increased significantly since 1980. This increase was mainly owing to an increase in cerebral palsy and psychiatric disorder prevalence rates. CONCLUSION: Future research aimed to explain these trends over time can be based on such data. The present knowledge is useful for planning purposes. PMID- 12538311 TI - Resuscitation training of paediatricians. AB - Intensive, didactic courses teaching resuscitation training are currently not only very popular, but are also required curricula content for certain aspects of medical training. There are a number of such courses available in the field of paediatrics, each with a different emphasis and target audience. They are not inexpensive, as they utilise large amounts of training equipment and have a high instructor to student ratio. This review will examine the course structures, instructor training, the need for such courses, and effect they may have on patient outcome. PMID- 12538312 TI - Acute disseminated encephalomyelitis: recognition in the hands of general paediatricians. AB - Acute disseminated encephalomyelitis will often present to the general paediatrician as an acute polysymptomatic encephalopathy, and initially the diagnosis may not be clear. A brain MRI scan is essential in establishing the diagnosis and so enabling appropriate advice and treatment to be given. Multicentre clinical audit of outcome and controlled therapeutic trials are needed to secure an evidence base for current practice. PMID- 12538313 TI - A seven year experience of medical emergencies in the assessment unit. AB - AIMS: To analyse retrospectively all referrals to the assessment unit during a seven year period, to determine their sources and destination. METHODS: All referrals over the seven year period were analysed. Parental satisfaction was determined using a questionnaire in some of the patients. The disease pattern and the investigations performed were determined. The community nurses' working hours and type of work done were analysed. RESULTS AND CONCLUSIONS: A total of 43 496 children were seen in the unit. Over 65% of the patients were referred by the general practitioners; 13 517 (34.2%) of those referred to the unit were discharged directly from the unit. Respiratory disorders and gastrointestinal problems were commonly seen. The children discharged from the unit did not have significantly more tests done on them. Most of the parents whose children were discharged from the unit were happy to be managed at home. The community nurses attended many children who needed intravenous therapy and advice on fluid rehydration. Community nurses reduce admission to the wards by working with other members in the assessment unit. This in turn provides a single point of entry and bridges the gap between primary and secondary care. We suggest recommendations on setting up such a unit. PMID- 12538314 TI - Amitraz poisoning, an emerging problem: epidemiology, clinical features, management, and preventive strategies. AB - BACKGROUND: Amitraz is a pharmaceutical, veterinary, and agricultural product which is used worldwide under numerous generic names as an acaricide and insecticide. Because of its widespread use amitraz poisoning has come emerged as a cause of childhood poisoning during the past decade, particularly more in certain countries such as Turkey. AIMS AND METHODS: To report the clinical features, the management, and the preventive strategies of amitraz poisoning in nine children, and review the previously reported 137 cases in humans. RESULTS: Five male and four female children aged 10 months to 8 years were admitted to our department. The estimated ingested dose ranged between 89.2 and 163 mg/kg and estimated time from ingestion to presentation was 30-120 minutes. The initial signs and symptoms were impaired consciousness, drowsiness, vomiting, disorientation, miosis, mydriasis, hypotension, bradycardia, tachypnoea, hypothermia, and generalised seizures. Hyperglycaemia, glycosuria, and minimal increase in transaminase levels were observed. None required mechanical ventilation. CNS depression resolved spontaneously within 4-28 hours in all. The length of hospital stay was two to three days; all had a good outcome. CONCLUSION: This review details preventive measures and management strategies of amitraz poisoning, including the importance of following patients closely in the intensive care unit, monitoring their respiratory, cardiovascular, and central nervous systems since they may occasionally experience serious cardiopulmonary side effects. PMID- 12538315 TI - Lung function and respiratory health in adolescents of very low birth weight. AB - AIMS: To determine if very low birth weight (VLBW; birth weight <1500 g) is associated with reduced lung function and respiratory health in adolescence and, if it is, whether this impairment is associated with prematurity or intrauterine growth restriction. METHODS: A geographically defined cohort of 128 VLBW infants and an age, sex, and school matched comparison group born in 1980/81 were studied. The cohort and comparison group were assessed at 15 years of age. The birth weight ratio of the index cases (observed birth weight/expected birth weight for the gestation) was determined to assess the degree of growth restriction. Respiratory support received during the neonatal period was obtained from hospital records. Smoking habits and respiratory morbidity were obtained through questionnaires. Forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), and forced expiratory flow when 25-75% of FVC is expired (FEF(25 75%)) were measured using a portable spirometer. The values are expressed as percentage predicted for height, age, and gender using standard reference values. Adjustments were made for smoking habits of mother and children. RESULTS: The differences in means between index and comparison groups for FEF(25-75%) ( 12.42%; p < 0.001) and FEV1/FVC (-3.53%; p < 0.001) ratio were statistically significant. The differences in FVC and FEV1 were not significant. No correlation was found between the birth weight ratio and lung function among the index cohort. Chronic cough, wheezing, and asthma were more common among the index cohort than in the comparison group. Within the index group, there was no difference in lung function between those who received and those who did not receive respiratory support. CONCLUSION: Adolescents who were VLBW compared with matched controls showed medium and small airways obstruction. This was associated with prematurity rather than intrauterine growth restriction or having received respiratory support during the neonatal period. The index VLBW cohort compared with their controls were also more prone to chronic cough, wheezing, and asthma. PMID- 12538317 TI - Blood pressure in sleep disordered breathing. AB - AIMS: To investigate blood pressure (BP) in children with sleep disordered breathing (SDB). METHODS: BP was measured during single night polysomnography in 23 suspected SDB child patients with adenotonsillar hypertrophy, but without respiratory or heart failure, or coma. The age related changes of the observed BP were normalised to the BP index. The BP indices were examined in relation to SDB measures, such as the desaturation time (percentage of time with oxygen saturation (SaO2) <90% against the total sleep time), SaO2 nadir, apnoea hypopnoea index (AHI), and arousal index, in addition to age and body mass index (BMI). RESULTS: The systolic BP index during rapid eye movement sleep (REMS) tended to correlate with AHI, while the diastolic index during REM sleep showed a significant correlation with AHI. The BP indices during non-REMS and wakefulness showed no correlation with the parameters obtained. Patients with an AHI of 10 or more (n = 7, AHIhigh) had significantly higher systolic and diastolic BP indices during both wakefulness and REMS, compared with those with an AHI of less than 10 (n = 16, AHIlow). Two patients in AHIhigh showed no sleep related dip of diastolic BP, and three patients in AHIlow lacked the sleep related dip in systolic BP. By means of multiple regression analysis, age, BMI, and AHI were found to be significant predictor variables of the systolic BP index during REMS. CONCLUSIONS: BP in paediatric SDB patients is positively correlated with the degree of SDB. PMID- 12538318 TI - Partial splenectomy in cystic fibrosis patients with hypersplenism. AB - We report three cystic fibrosis (CF) patients with hypersplenism who underwent partial splenectomy. The postoperative course was uneventful in two patients; one patient developed a complication necessitating resection of the rest of the spleen. Haematological parameters improved and oesophageal varices regressed in all patients. On follow up, one patient showed a normal spleen, the other a normally functioning accessory spleen; the third patient again developed splenomegaly with hypersplenism. Partial splenectomy is a promising therapeutic option for CF patients with hypersplenism. PMID- 12538320 TI - Attentional ability among survivors of leukaemia treated without cranial irradiation. AB - BACKGROUND: Previous research has indicated that children who have received treatment for leukaemia which includes cranial irradiation exhibit deficits in their ability to focus attention. It has been suggested that the use of cranial irradiation may have a role to play in long term sequelae. AIMS: To investigate neuropsychological functioning among children treated for leukaemia without cranial irradiation. METHODS: In a cross sectional study, 17 leukaemic patients and their sibling controls were assessed using a neuropsychological model of attention. All were treated on the UKALL XI protocol and none had received cranial irradiation. Participants completed the Arithmetic subtest and Digit Span subtest of the Weschler Intelligence Scale for Children-Revised to assess focus encode elements of attention; the Coding subtest and the Speed of Information subtest of the BAS to assess focus-execute aspects of attention; the VIGIL computerised battery to assess sustain elements of attention; and the Wisconsin Card Sorting test to assess the ability to shift attention. RESULTS: These children did not exhibit the deficits witnessed in previous cohorts, and were performing at comparable levels to their controls on all measures of attention CONCLUSIONS: These findings suggest that children who have received treatment for leukaemia without the use of cranial irradiation do not show the neuropsychological insult found in earlier treatment groups. PMID- 12538321 TI - Secular trends in growth in diabetes: are we winning? AB - AIM: To determine potential effects of modern treatment on growth in diabetic children. METHODS: Retrospective analysis of growth in diabetic children stratified by their year of diagnosis between 1974 and 1995. A total of 451 children and adolescents attending the Diabetes Outpatient and Outreach Clinics of Royal Alexandra Hospital for Children in Sydney and rural NSW, Australia were studied. Standard deviation scores (SDS) for height and body mass index (BMI) were assessed at diagnosis, five years later (n = 451), and 10 years later (n = 111). RESULTS: After five years of diabetes duration height SDS loss correlated with higher HbA(1c) and fewer injections. BMI SDS gain correlated with HbA(1c) and age at diagnosis. Although there was no significant difference in their height SDS or age at diagnosis, children diagnosed 1974-90 were significantly shorter than children diagnosed 1991-95 (height SDS 0.07 v 0.37) after five years diabetes duration. Furthermore, over 5 and 10 years, the 1979-90 group had lost significant height SDS (mean change -0.20 at 5 years, -0.29 at 10 years); this did not occur in the 1991-95 group (-0.01 at 5 years, -0.13 at 10 years). The BMI SDS increased significantly after 10 years in the 1974-90 group (mean change 0.37) but not in the 1991-95 group. There was no significant difference in the 174 females' age of menarche (13.0 v 12.8 years). CONCLUSIONS: Children with diabetes treated with modern regimens maintain their increased height from diagnosis better, and after five years diabetes duration, were taller than children diagnosed before 1991. PMID- 12538322 TI - The incidence of hypoglycaemia in children with type 1 diabetes and treated asthma. AB - AIMS: To investigate whether treatment of coexisting asthma has any effect on the incidence of hypoglycaemia and on glycaemic control in children with type 1 diabetes. METHODS: An observational study of children attending the paediatric diabetes clinics of five hospitals in the North Trent Region. Information on the frequency of hypoglycaemia in the preceding three months, treatment for asthma, and the individual's latest HbA1c, was recorded when they attended for review. RESULTS: Data were collected on 226 children, of whom 27 (12%) had treated asthma. Only 11/27 children with asthma were taking their prescribed inhaled steroids. All used beta agonists at least once a week. There was a reduction of 20% in the incidence of hypoglycaemia in the diabetic children with treated asthma. Of the children with diabetes and treated asthma, 52% reported an episode of hypoglycaemia in the previous three months compared to 72% of those with only diabetes. There was no difference in the proportion of children experiencing nocturnal or severe hypoglycaemia. Although not significant, those with asthma and diabetes also had better overall control (HbA1c 8.8%) compared to those with diabetes alone (HbA1c 9.3%). CONCLUSIONS: Diabetic children with treated asthma have significantly fewer episodes of hypoglycaemia and better glycaemic control compared to children with diabetes alone. This observation needs further investigation but raises an interesting question. Do the drugs used to treat asthma, in particular beta agonists, have the therapeutic potential to reduce hypoglycaemia and facilitate an improvement in glycaemic control? PMID- 12538323 TI - Sudden infant death due to disseminated pneumococcal infection. PMID- 12538324 TI - Successful radioiodine treatment in a 3 year old child with Graves' disease following antithyroid medication induced neutropenia. AB - A 3 year old child with Graves' disease and mitral valve prolapse became neutropenic on carbimazole therapy. She was switched to propylthiouracil but the neutropenia recurred. She was treated with radioiodine but required two doses of 113 MBq and then 198 MBq five months later before becoming hypothyroid. The mitral valve prolapse resolved when she was euthyroid on thyroxine replacement. Antithyroid drugs, surgery, and radioiodine all have a place in the management of the thyrotoxic child. PMID- 12538325 TI - An unusual presentation of perinatally transmitted hepatitis C. AB - A unique presentation of perinatally acquired hepatitis C (HCV) with acute jaundice and chronic aggressive liver disease in a previously asymptomatic preadolescent is described. The difficulties in establishing the diagnosis and the importance of confirmatory testing for HCV in cryptogenic liver disease are discussed. PMID- 12538326 TI - Ganglioneuroblastoma presenting as dilated cardiomyopathy. AB - We report an unusual presentation of ganglioneuroblastoma with features of dilated cardiomyopathy in a 22 month old girl. She was admitted with cardiomegaly; during echocardiography a suspicious abdominal mass was detected by chance. Further imaging studies, including abdominal ultrasonography and spiral computed tomography, revealed a solid mass originating in the right adrenal gland. Metabolic studies and pathological findings were compatible with ganglioneuroblastoma. Following tumour removal and supportive therapy for cardiomyopathy, her clinical condition and laboratory findings improved. Although ganglioneuroblastoma with features of dilated cardiomyopathy is rare, because neurogenic tumours may be involved in its development, measurement of catecholamines in children with dilated cardiomyopathy is strongly recommended. PMID- 12538327 TI - Radial artery pseudoaneurysm successfully treated by compression bandage. AB - In children, surgery for radial artery pseudoaneurysm (PA) may be followed by growth retardation of the hand because of inadequate blood flow. We believe this is the first report of a child with PA of the radial artery cured by compression bandage. Conservative management is a safe and valuable initial treatment option for uncomplicated radial PA. PMID- 12538328 TI - Fractured fibula can mimic irritable hip. PMID- 12538330 TI - Importance of timing of risk factors for cerebral oedema during therapy for diabetic ketoacidosis. AB - Cerebral oedema is the most common cause of mortality and morbidity during the first day of conventional treatment for diabetic ketoacidosis in paediatric patients. It is possible that therapy contributes to its development. Risk factors that predispose to cerebral oedema should lead to an expansion of the intracellular and/or the extracellular fluid compartment(s) of the brain because water normally accounts for close to 80% of brain weight. With respect to the intracellular fluid compartment, the driving force to cause cell swelling is a gain of effective osmoles in brain cells and/or a significant decline in the effective osmolality of the extracellular fluid compartment. Factors leading to an expansion of the intracerebral extracellular fluid volume can be predicted from Starling forces acting at the blood-brain barrier. Some of these risk factors have an early impact, while others have their major effects later during therapy for diabetic ketoacidosis. Based on a theoretical analysis, suggestions to modify current therapy for diabetic ketoacidosis in children are provided. PMID- 12538331 TI - BK virus DNA in CSF of immunocompetent and immunocompromised patients. AB - AIM: To investigate the possible aetiological role of BK and JC viruses in immunocompetent and immunocompromised children with suspected encephalitis and meningoencephalitis. METHODS: The polymerase chain reaction and microplate hybridisation method was employed for the detection of polyomavirus DNA in 266 CSF specimens collected from immunocompetent and immunocompromised patients. RESULTS: BK virus DNA was detected in three (2.1%) CSF samples taken from patients aged 2-5 years; two were patients with acute lymphocytic leukaemia without overt neurological symptoms, the other was a patient with suspected encephalitis. BK virus DNA was also detected in two (1.6%) CSF samples taken from older children in the age range 10-16 years; both children had suspected encephalitis. JC virus DNA was not found in any CSF sample from either age group. CONCLUSIONS: Detection of BK virus in the CSF of immunocompromised and immunocompetent patients with suspected neurological disease suggests that this virus may have had a pathogenic role in the aetiology of this condition. PMID- 12538332 TI - On archimedes. PMID- 12538333 TI - Conflicting advice. PMID- 12538334 TI - LP and Glasgow coma score. PMID- 12538335 TI - Are interhospital transport teams de-skilling the DGH paediatricians? PMID- 12538336 TI - More lumbar punctures, please! PMID- 12538337 TI - Acute ataxia complicating Langerhans cell histiocytosis. PMID- 12538338 TI - A self-fulfilling prophesy? PMID- 12538339 TI - The relationship between Helicobacter pylori infection and iron deficiency: seroprevalence study in 937 pubescent children. PMID- 12538340 TI - Hypocalcaemia and calcitonin precursors in critically ill patients. PMID- 12538342 TI - Cancer as an evolutionary process at the cell level: an epidemiological perspective. AB - Germ-line mutations (present in all cells) in genes that are crucial for the cell cycle cause cancer only in specific cell lines (e.g. mismatch repair genes in the colon; BRCA1-2 in breast and ovary; other cancers in Bloom syndrome, neurofibromatosis and xeroderma pigmentosum). The mutation rate of genes other than mismatch repair or p53 is the same in colon cancer and in normal cells, indicating that a 'mutator phenotype', increasing the rate of mutations in many genes, is not an essential feature of sporadic cancers; conversely, fusion genes, TEL-AML1/AML1-ETO, typical of leukemia, are 100 times more frequent at birth than in overt leukemia in children, indicating that further selective events are needed to cause malignancy. The devastating impairment of immunity, as in AIDS patients, does not cause cancer other than Kaposi's sarcoma and non-Hodgkin's lymphoma, although immunological control is considered to be an essential mechanism in preventing the spread of cancer cells. These observations suggest that cell-specific additional events are needed to explain carcinogenesis. Carcinogenesis has been traditionally interpreted as the sequence of initiation (mutation) and promotion (clone expansion), with an interesting similarity with the neo-Darwinian theory of evolution, based on a first stage of genetic change (including recombination) and a second stage of selection. I propose that carcinogenesis consists in two general phases (not necessarily stages), i.e. genetic change followed by clone expansion (selective advantage). As in neo Darwinian theory selection is chiefly represented by the elimination of the less fit, the selection of mutated cells would mainly consist in resistance to apoptosis or other types of 'bottlenecks' that hamper a cell's survival; an example of such a bottleneck is the autoimmunity that induces paroxysmal nocturnal hemoglobinuria in individuals with PIG-A mutations. Cancer rates show great variation in different countries around the world, a variation only marginally explained by genetic differences. More interestingly, migrants change their risk of cancer by adapting to that of the population into which they move: as these changes are not likely to be entirely due to mutagens in the environment, we have to invoke selective pressure over mutated cells to explain them. My theory is that mutated cells adapt to environmental 'niches' better than normal cells, in a 'gene-environment interaction' that involves the history of the genetic changes the cell has undergone and the kind of environment in which it happens to live. PMID- 12538343 TI - Short-term depletion of catalase suppresses cadmium-elicited c-Jun N-terminal kinase activation and apoptosis: role of protein phosphatases. AB - The c-Jun N-terminal kinase (JNK) is a vital stress-activated signal that can be regulated differentially under oxidant or antioxidant conditions. Recently, we have reported that activation of JNK by cadmium chloride (Cd) contributes to apoptosis in CL3 human lung adenocarcinoma cells. Although oxidative stress has been implicated in numerous biochemical effects altered by Cd, its role in Cd elicited JNK activation has not been established. Here we report that catalase is crucial for the activation of JNK by Cd. Short-term treatment of 3-amino-1,2,4 triazole (3AT), a specific catalase inhibitor, completely suppressed the Cd elicited JNK activation, conversely, exogenous addition of catalase increased the intensity and duration of JNK activation in Cd-treated CL3 cells. Co administering high doses of H(2)O(2) (500-1000 micro M) with Cd also markedly decreased JNK activity, although at doses <200 micro M H(2)O(2) enhanced the Cd elicited JNK activation in CL3 cells. 3AT also blocked JNK activation in Cd treated normal human fibroblasts and Chinese hamster ovary cells, and in UV irradiated CL3 cells. However, mannitol, a hydroxyl radical scavenger, did not alter the JNK activity in Cd-treated human and rodent cells. Intriguingly, sodium fluoride or okadaic acid, inhibitors for serine/threonine protein phosphatases (PP), recovered the JNK activity in CL3 cells exposed to Cd plus 3AT; however, the protein tyrosine phosphatases inhibitor sodium orthovanadate did not. Furthermore, 3AT decreased but catalase increased the Cd-induced cytotoxicity, apoptosis and procaspase-3 degradation in CL3 cells. Together, these results indicate that persistent activation of apoptotic JNK signal by Cd requires functional catalase and that short-term depletion of catalase activity may facilitate okadaic acid-sensitive PP to down-regulate the JNK activation and may predispose these cells to carcinogenic transformation upon Cd exposure. PMID- 12538344 TI - Human DNA methyltransferase gene DNMT1 is regulated by the APC pathway. AB - Epigenomic changes in DNA methylation patterns are evident in a variety of cancers, including colorectal cancer (CRC). In addition, a large proportion of CRC tumors and cell lines harbor genetic mutations in the APC/beta-catenin/TCF transcription activation pathway. While several target genes have been proposed, a causal downstream agent between APC mutation and cancer has not been fully established. Because previous work implicates DNA methyltransferase (DMNT1) as a critical point in tumorigenesis and recent studies suggest that familial CRC also exhibits epigenetic alterations, we sought to investigate whether this gene might be regulated by APC in CRC. Reconstitution of wild type APC in HT-29 CRC cell lines reduced the expression of both a reporter gene driven by the minimal DNMT1 promoter and DNMT1 mRNA that is independent of cell growth stasis. We also provide evidence for a causal role of DNMT1 in CRC by demonstrating that antisense-driven reduction of DNMT1 mRNA inhibits anchorage-independent growth, an indicator of tumorigenesis, of CRC cells. These data support future consideration of DNMT1 as a target in the treatment of CRC. PMID- 12538346 TI - Assessment of mismatch repair function in leukaemic cell lines and blasts from children with acute lymphoblastic leukaemia. AB - Defects in the DNA mismatch repair (MMR) pathway have recently been shown to be associated with resistance to several of the cytotoxic drugs used in the treatment of children with acute lymphoblastic leukaemia (ALL). We have assessed the MMR status of a range of leukaemic cell lines using an in vitro repair assay and correlated this with protein expression of the best characterized components of the system. We have also assessed MMR in leukaemic blasts from a limited panel of children with ALL and related this to Ki67 expression as a measure of proliferative capacity. Out of nine leukaemic cell lines tested, five of the seven lymphoid lines showed little or no repair using the in vitro assay and had low MMR protein expression. In three (NALM-6, Reh and MOLT 4) MMR defects have not been previously reported. Immunohistochemistry of clinical samples showed a wide range of expression of MLH1, MSH2 and Ki67 in nine cases studied at presentation, with a highly statistically significant correlation between MLH1 and Ki67 expression (r(2) = 0.96, P < 0.0001, Pearson correlation). Western blotting demonstrated high expression of MLH1, PMS2, MSH2 and MSH6 proteins. In vitro analysis of G.T repair using lymphoblast cytosol from the same patients showed a wide range of proficiency, which was markedly reduced in one case studied at relapse. These results suggest that MMR defects are more common in leukaemic cell lines and acute lymphoblastic leukaemias than previously thought. PMID- 12538345 TI - Hypermethylation of metallothionein-3 CpG island in gastric carcinoma. AB - The expression of metallothionein (MT)-3 is often markedly reduced in gastric carcinoma (GC). The molecular mechanism of this MT-3 downregulation is unknown. Transcriptional silencing of MT-3 by methylation of CpG island was investigated by nucleotide sequencing and denaturing high performance liquid chromatography (DHPLC) analyses. We found that normal brain tissue and a xenografted GC that expressed MT-3 mRNA had unmethylated regions of the CpG island in intron1 of this gene. On the other hand, gastric cancer cell lines AGS and MKN445, a xenografted GC, and a representative primary gastric cancer that had no expression of MT-3 mRNA demonstrated hypermethylation of the MT-3 intron1 CpG island. Treatment of the gastric cancer cell lines with 5-azacytidine resulted in new expression of MT 3 mRNA in these cells. A quantifying DHPLC assay was developed to determine the methylation status of this specific region of the MT-3 gene. Fifty-eight primary GC and their corresponding normal gastric epithelial tissues, and 34 normal gastric mucosa were analyzed for MT-3 methylation by DHPLC in the region of methylation abnormalities initially identified. Our DHPLC analyses of the methylated MT-3 product demonstrated that the primary gastric cancers have an average methylation percentage of 6.3% per tumor compared with 2.4% in normal gastric tissues (P < 0.05). The MT-3 was not methylated in all of eight P53 positive GCs and hypermethylated in eight of 13 P53-negative cases by immunohistochemistry staining (P = 0.007). In conclusion, the CpG island in the MT-3 intron1 are abnormally hypermethylated in many gastric carcinomas and may account for the downregulation of MT-3 in gastric carcinogenesis. PMID- 12538347 TI - Impact of Dnmt1 deficiency, with and without low folate diets, on tumor numbers and DNA methylation in Min mice. AB - Although a number of studies have suggested that diets with low intake of folate, an important methyl donor, are associated with increased risks of colon cancer and its precursor the adenomatous polyp, the underlying mechanisms are poorly understood. Dysregulation and instability of DNA methylation and alterations in the levels of the predominant DNA methylating enzyme, DNA (cytosine-5) methyltransferase 1 (Dnmt1), have also been linked to tumorigenesis. We have used a combination of genetic and dietary manipulation to assess the effects of reduced Dnmt1 expression with and without folate deficiency on tumor induction in the Apc(Min) mouse. Apc(Min) mice with a reduction in Dnmt1 expression (Apc(Min/+)/Dnmt1(C/+)) had significantly lower tumor numbers than Apc(Min) mice with normal Dnmt1 (Apc(Min/+)/Dnmt1(+/+)). Dietary folate deficiency from weaning to 13 weeks of age did not affect tumor number or size in Apc(Min/+)/Dnmt(+/+) mice. However, in Apc(Min/+)/Dnmt1(C/+) mice with high baseline tumor numbers (41 +/- 4), folate deficiency was associated with a decreased absolute number of tumors (27 +/- 3), but a higher proportion of larger tumors as compared with mice on the control diet. In the repeat experiment, Apc(Min/+)/Dnmt1(C/+) mice had low baseline tumor numbers (20 +/- 2) and folate deficiency did not affect tumor number (23 +/- 4) or size as compared with the same mice on the control diet. These results suggest that, in the presence of Dnmt1 deficiency, the effects of folate deficiency on tumor number and size may depend on the stage of adenoma development when folate deficiency is initiated. We also show that folate deficiency with or without reductions in Dnmt1 did not affect overall genomic DNA methylation or the methylation levels of two candidate genes, E-cadherin or p53, in normal or neoplastic intestinal tissue. In conclusion, genetic deficiency in Dnmt1 with or without folate deficiency decreases tumor number in the Apc(Min) mouse model, but this effect may not be mediated by changes in SAM or SAH levels, nor by alterations in global methylation in the pre-neoplastic intestinal tissue. PMID- 12538348 TI - Frequent hypermethylation of the 5' CpG island of the mitotic stress checkpoint gene Chfr in colorectal and non-small cell lung cancer. AB - Chfr, a mitotic stress checkpoint gene, regulates a prophase delay in cells exposed to agents that disrupt microtubules, such as nocodazole and taxol. In the present study, we report that Chfr is frequently methylated in cell lines derived from tumors of the colon (80%), brain (100%) and bone (100%). In addition, Chfr was methylated in 37% of primary colon adenocarcinomas and in 10% of primary non small cell lung carcinomas. In normal colon tissue, but not lung, there was evidence for age-related methylation of Chfr, suggesting that in some cases the tumor may have arisen from a methylated clonal precursor. Methylation was associated with loss of Chfr mRNA and protein expression in cancer cell lines. In cells with methylated Chfr, treatment with the demethylating agent 5-aza-2' deoxycytidine resulted in re-expression of Chfr, and partial restoration of the prophase checkpoint. These results suggest that epigenetic inactivation of Chfr may be responsible for many of the checkpoint defects observed in human cancers. PMID- 12538349 TI - Persistent activation of ERK1/2 by lead acetate increases nucleotide excision repair synthesis and confers anti-cytotoxicity and anti-mutagenicity. AB - Lead, a possible human carcinogen, affects signal transduction pathways in many aspects, yet exhibits low mutagenicity in human cells. In this study, we explore whether signaling pathways including the four MAPKs and AKT affect DNA repair and mutagenicity in the exposure of mammalian cells to lead acetate [Pb(II)]. Pb(II) increased the phosphorylated ERK1/2 and phosphorylated AKT but not the phosphorylated ERK5, phosphorylated p38 and JNK activity in human non-small cell lung adenocarcinoma CL3 cells. The duration of ERK1/2 activation was much longer than AKT activation and these two signals were independently activated by Pb(II) in CL3 cells. Intriguingly, a MKK1/2 inhibitor PD98059 (25-50 micro M) markedly suppressed ERK1/2 activation and greatly promoted the hprt mutation frequency and cytotoxicity in Pb(II)-treated CL3 cells. Conversely, inhibition of the AKT signal by wortmannin did not exhibit such effects. Inhibition of the persistently activated ERK1/2 in Pb(II)-treated diploid human fibroblasts by PD98059 also markedly increased the mutagenicity and cytotoxicity. The Pb(II)-induced mutagenicity and cytotoxicity were significantly higher in nucleotide excision repair (NER)-deficient UVL-10 rodent cells than their counterpart AT3-2 cells; also, ERK1/2 activation by Pb(II) was observed in AT3-2 but not UVL-10 cells. Furthermore, cellular NER synthesis was enhanced by Pb(II) exposure, which was markedly suppressed by PD98059. Activation of ERK1/2 by expressing a constitutively active form of MKK1 in CL3 cells also elevated cellular NER synthesis. Together, these results indicate that persistent activation of ERK1/2 signaling by Pb(II) enhances cellular NER synthesis, thereby conferring anti cytotoxicity and anti-mutagenicity. PMID- 12538350 TI - Occupational exposure to heavy metals: DNA damage induction and DNA repair inhibition prove co-exposures to cadmium, cobalt and lead as more dangerous than hitherto expected. AB - Co-exposure to cadmium, cobalt, lead and other heavy metals occurs in many occupational settings, such as pigment and batteries production, galvanization and recycling of electric tools. However, little is known about interactions between several heavy metals. In the present study we determined DNA single strand break (DNA-SSB) induction and repair capacity for 8-oxoguanine in mononuclear blood cells of 78 individuals co-exposed to cadmium (range of concentrations in air: 0.05-138.00 micro g/m(3)), cobalt (range: 0-10 micro g/m(3)) and lead (range: 0-125 micro g/m(3)). Exposure to heavy metals was determined in air, blood and urine. Non-parametric correlation analysis showed a correlation between cadmium concentrations in air with DNA-SSB (P = 0.001, R = 0.371). Surprisingly, cobalt air concentrations correlated even better (P < 0.001, R = 0.401), whereas lead did not correlate with DNA-SSB. Logistic regression analysis including 11 possible parameters of influence resulted in a model showing that cobalt in air, cadmium in air, cadmium in blood and lead in blood influence the level of DNA-SSB. The positive result with cobalt was surprising, since exposure levels were much lower compared with the TRK-value of 100 micro g/m(3). To examine, whether the positive result with cobalt is stable, we applied several logistic regression models with two blocks, where all factors except cobalt were considered preferentially. All strategies resulted in the model described above. Logistic regression analysis considering also all possible interactions between the relevant parameters of influence finally resulted in the following model: Odds ratio = 1.286(Co in air) x 1.040(Cd in air) x 3.111(Cd in blood) x 0.861(Pb in air) x 1.023(Co in air x Pb in air). This model correctly predicts an increased level of DNA-SSB in 91% of the subjects in our study. One conclusion from this model is the existence of more than multiplicative effects for co-exposures of cadmium, cobalt and lead. For instance increasing lead air concentrations from 1.6 to 50 micro g/m(3) in the presence of constant exposures to cobalt and cadmium (8 micro g/m(3) and 3.8 micro g/m(3)) leads to an almost 5 fold increase in the odds ratio, although lead alone does not increase DNA-SSB. The mechanism behind these interactions might be repair inhibition of oxidative DNA damage, since a decrease in repair capacity will increase susceptibility to reactive oxygen species generated by cadmium or cobalt. Indeed, repair of 8 oxoguanine decreased with increasing exposures and inversely correlated with the level of DNA-SSB (P = 0.001, R = -0.427). Protein expression patterns of individuals exposed to cobalt concentrations of approximately 10 micro g/m(3) were compared with those of unexposed individuals using two-dimensional gel electrophoresis. Qualitative and apparent quantitative alterations in protein expression were selective and certainly occurred in <0.1% of all proteins. In conclusion, the hazard due to cobalt exposure - that has been classified only as IIB by the IARC - seems to be underestimated, especially when individuals are co exposed to cadmium or lead. Co-exposure may cause genotoxic effects, even if the concentrations of individual heavy metals do not exceed TRK-values. PMID- 12538351 TI - Apoptogenic effects of black tea on Ehrlich's ascites carcinoma cell. AB - Next to water, tea is the most ancient and widely consumed beverage in the world. Epidemiological studies have suggested a cancer protective effect, but the results obtained so far are not conclusive. In the current study, mechanisms of the apoptogenic effect of black tea extract were delineated. Black tea administration to Ehrlich's ascites carcinoma (EAC)-bearing Swiss albino mice caused a significant decrease in the tumor cell count in a dose-dependent manner. Flowcytometric analysis showed an increase in the number of cells in the sub G(0)/G(1) population signifying tumor cell apoptosis by black tea. These results were further confirmed by nuclear staining that demonstrated distinct morphological features of apoptosis. Our data also revealed an increase in the expression of pro-apoptotic protein p53 in EAC. It is known that upon p53 induction, multiple downstream factors contribute to the decision making between growth arrest and apoptosis. Among those, pro-apoptotic gene Bax is up regulated during p53-mediated apoptosis. On the other hand, p53-mediated growth arrest involves p21 as a major effecter. In our system, increase in p53 expression was followed by moderate expression of p21/Waf-1 and high expression of Bax at protein levels. Interestingly, anti-apoptotic protein Bcl-2 was down regulated resulting in decrease in Bcl-2/Bax ratio. All these observations together signify that black tea-induced apoptogenic signals overrode the growth-arresting message of p21, thereby leading the tumor cells towards death. PMID- 12538352 TI - The effect of dietary folate on genomic and p53-specific DNA methylation in rat colon. AB - Folate is an important mediator in the transfer of methyl groups for DNA methylation, abnormalities of which are considered to play an important mechanistic role in colorectal carcinogenesis. This study investigated the time dependent effects of dietary folate on genomic and p53 (in the promoter region and exons 6-7) DNA methylation in rat colon, and how these changes are related to steady-state levels of p53 transcript. Despite a marked reduction in plasma and colonic folate concentrations, a large increase in plasma homocysteine (an accurate inverse indicator of folate status), and a progressive decrease in colonic S-adenosylmethionine (SAM; the primary methyl donor for methylations) to S-adenosylhomocysteine (SAH; a potent inhibitor of methylations) ratio, isolated folate deficiency did not induce significant genomic DNA hypomethylation in the colon. Paradoxically, isolated folate deficiency increased the extent of genomic DNA methylation in the colon at an intermediate time point (P = 0.022). Folate supplementation did not modulate colonic SAM, SAH and SAM to SAH ratios, and genomic DNA methylation at any time point. The extent of p53 methylation in the promoter and exons 6-7 was variable over time at each of the CpG sites examined, and no associations with time or dietary folate were observed at any CpG site except for site 1 in exons 6-7 at week 5. Dietary folate deprivation progressively decreased, whereas supplementation increased, steady-state levels of p53 transcript over 5 weeks (P < 0.05). Steady-state levels of p53 mRNA correlated directly with plasma and colonic folate concentrations (P = 0.41-0.49, P < 0.002) and inversely with plasma homocysteine and colonic SAH levels (r = 0.37-0.49, P < 0.006), but did not significantly correlates with either genomic or p53 methylation within the promoter region and exons 6-7. The data indicate that isolated folate deficiency, which significantly reduces steady-state levels of colonic p53 mRNA, is not associated with a significant degree of genomic or p53 DNA hypomethylation in rat colon. This implies that neither genomic or p53 hypomethylation within exons 6-7 nor aberrant p53 methylation within the promoter region is likely a mechanism by which folate deficiency enhances colorectal carcinogenesis in the rat. PMID- 12538353 TI - beta-Catenin mutation is selected during malignant transformation in colon carcinogenesis. AB - Activating mutations in the beta-catenin gene is thought to be responsible for the excessive beta-catenin signaling involved in the majority of colon carcinomas in rodent models. Our recent study which indicated that beta-catenin mutations are present frequently in early dysplastic lesions of rat colon induced by a colon-specific carcinogen, azoxymethane led us to perform more specifically a comparative study regarding types of the beta-catenin mutation as well as K-ras mutations between such early appearing lesions and colon tumors. Male F344 rats, 6 weeks old, received s.c. injections of azoxymethane (15 mg/kg body weight) once a week for 3 weeks, and were killed at 16 and 46 weeks of age. Colons of animals killed at 16 weeks of age were processed for early altered lesions. Colon tumors from animals killed at 46 weeks of age were evaluated histopathologically. Laser capture microdissection system was used to obtain DNA of epithelial cells in both intramucosal lesions and colon tumors. After amplification of exon 3 of the beta catenin gene and exon 1 of the K-ras gene, the products were then sequenced directly in both directions. Mutations in the exon 3 of beta-catenin gene were detected in 22 of 56 early lesions (39.3%) and 21 of 37 colon cancers (56.8%). Remarkably, all beta-catenin mutations detected in the colon tumors converged at codons encoding functionally important residues that may directly mediate beta catenin degradation, whereas mutations in the early appearing lesions were found to be scattered in the exon 3 of the gene. K-ras mutations were also detected in 24 of 56 early lesions (42.9%) and 11 of 37 colon cancers (29.7%). All K-ras mutations converged at codon 12 and codon 13, even in the early lesions. The results of this study provide evidence for the first time that beta-catenin mutation is selected during the colon carcinogenesis. Our results also suggest that the activation of beta-catenin signaling pathway is not only an initiating event, but also plays a pivotal role in the promotion stage of colorectal carcinogenesis. PMID- 12538354 TI - Affinity with Raf is sufficient for Ras to efficiently induce rat mammary carcinomas. AB - The role of three major Ras downstream effector pathways in the induction of mammary cancer was studied using an in situ mammary ductal gene delivery model. Replication-defective retroviral vectors were used to infect endogenous rat mammary epithelial cells with three individual Ras effector loop mutants, each of which transduces its signal through a different Ras effector pathway (Raf, PI3K or RalGDS). Several groups have used Ras effector loop mutants in cultured cells, clearly characterizing the signaling specificity of each over a wide range of cell lines and conditions. Each of the three Ras effector loop mutations impairs Ras for neoplastic transformation of immortal cell lines in culture. In contrast, when evaluated in vivo by infecting endogenous rat mammary epithelial cells in situ with retroviral vectors, we find that codon 12 mutant activated V12-Ras and all three V12-Ras effector loop mutants individually induce mammary carcinomas. Most notably, a Ras effector loop mutant that lacks affinity with PI3K and RalGDS but retains affinity with Raf (E38-V12-Ras) is relatively similar in potency to V12-Ras for mammary carcinoma induction. Two other Ras effector loop mutants, each lacking affinity with Raf, one retaining affinity with PI3K (C40-V12-Ras), the other with RalGDS (G37-V12-Ras), resulted in much longer tumor latency than E38-V12-Ras and V12-Ras and a reduced carcinoma frequency. Tumor latencies for V12-Ras, E38-V12-Ras, C40-V12-Ras and G37-V12-Ras were 4, 4, 11 and 12 weeks, respectively. We conclude that the Ras-Raf pathway can function independently of the Ras-PI3K and Ras-RalGDS pathways for rapid induction of rat mammary carcinomas, while Ras-PI3K and Ras-RalGDS pathways may also individually induce mammary carcinomas following a long latency. PMID- 12538355 TI - Azoxymethane-induced beta-catenin-accumulated crypts in colonic mucosa of rodents as an intermediate biomarker for colon carcinogenesis. AB - It is now well established that bile acids act as colon tumor promoters. However, a previous study provided conflicting data showing that dietary exposure of cholic acid (CHA), a primary bile acid, inhibits the carcinogen-induced formation of aberrant crypt foci (ACF), possible preneoplastic lesions, in colonic mucosa of rodents. Recently we found beta-catenin-accumulated crypts (BCAC) in colonic mucosa of rats initiated with azoxymethane (AOM) and provided evidence that BCAC might be preneoplastic lesions independent from ACF. In the present study, we investigated the modifying effects of dietary CHA on the formation of BCAC as well as ACF in male F344 rats after exposure to AOM to determine if the differences in the effect of CHA on these lesions could account for this discrepancy. The results indicate that administration of CHA (0.5%) in the diet during the post-initiation phase significantly reduced the total number, multiplicity and size of ACF (P < 0.00001) in AOM-exposed colonic mucosa as reported previously. The number of ACF even with >4 aberrant crypts/focus was also decreased significantly (P < 0.0002), suggesting that the large ACF are little resistant to continuous feeding of 0.5% CHA diet. Interestingly, the dietary CHA significantly enhanced both the multiplicity (P < 0.002) and size (P < 0.00001), but not the incidence, of AOM-induced BCAC when compared with the control diet group. Importantly, the number of large BCAC with >6 crypts/lesion was increased significantly by the dietary CHA (P < 0.003). Our results support the concept that BCAC are precursors of colon tumors and indicate the usefulness of BCAC as intermediate biomarkers for colon carcinogenesis, although the methodology for their detection requires further improvement. PMID- 12538356 TI - Simulated sunlight and benzo[a]pyrene diol epoxide induced mutagenesis in the human p53 gene evaluated by the yeast functional assay: lack of correspondence to tumor mutation spectra. AB - Many mutations in the p53 gene destroy the transcriptional transactivation function of the p53 protein. This function of p53 can be determined in a yeast assay using a p53 responsive reporter gene. The yeast assay could hold promise for the identification of mutagens implicated in human cancer if the p53 mutational spectra obtained with this assay would match human tumor mutation data. Ultraviolet (UV) light from the sun and polycyclic aromatic hydrocarbons, such as benzo[a]pyrene, are strongly implicated in the spectrum of p53 mutations found in human non-melanoma skin cancers and smoking-associated lung cancers, respectively. We have used these two model mutagens to assess the feasibility of using the p53 yeast assay in cancer epidemiology. After treatment of CpG methylated p53 DNA with a solar UV simulator or with benzo[a]pyrene diol epoxide (BPDE), the modified p53 sequences were assayed in yeast for mutational outcome. As expected, BPDE produced predominantly G to T transversions and simulated sunlight produced mostly C to T transitions at dipyrimidine sites in the p53 coding sequence. However, the preferentially mutated p53 sequences (hotspots) in the yeast assay were completely different from those in the mutational spectra found in human lung and skin cancers. The data indicate that this assay is not a reliable measurement of p53 mutagenesis in human tissues and that, perhaps, transcriptional activation is not the primary function of p53 in tumor suppression. PMID- 12538357 TI - Perturbations of the Ink4a/Arf gene locus in aflatoxin B1-induced mouse lung tumors. AB - Lung tumors from AC3F1 mice treated with aflatoxin B(1) (AFB(1)), were examined for loss of alleles, point mutations and hypermethylation of CpG sites within the promoters of the two genes in the Ink4a/Arf gene locus. Loss of microsatellite alleles in the Ink4a/Arf region occurred in 22 of 74 (30%) AFB(1)-induced lung tumors. Fifty-one of 61 (83%) tumors had at least partial methylation of CpG sites within the p16Ink4a promoter-exon 1alpha region. At least partial methylation of CpG sites was observed in 43 of 49 (88%) tumors analyzed for p19Arf promoter hypermethylation, with methylation of identified transcription factor binding sites or consensus sequences occurring in 21 tumors (DMP1/Ets in two tumors, CTCF in four tumors, E2F in three tumors, Sp1 in 16 tumors). Two tumors contained point mutations in the p19Arf promoter. Nuclear staining for p19(Arf) was decreased by 80-100% in 41 of 71 (58%) tumors. The concordance between p19Arf molecular perturbations and altered protein expression was 63%. However, upon comparing p19Arf promoter perturbations (i.e. methylation of functional transcription factor binding sites and point mutations) and altered p19(Arf) expression, the concordance was 86%, suggesting a mechanism for changes in protein expression in some tumors. There was an absence of a mutually exclusive relationship between disruption of p53 and p19(Arf), since the concordance was 62%. Similarly, no evidence was found of inverse relationships between perturbation of p16Ink4a and p19(Arf) (43% concordance) or p16Ink4(a) and p53 (37% concordance), suggesting that inactivation of these genes occurs independently and provides evidence that, although these genes may participate in cooperative cellular pathways, they also have functions in independent pathways that are important in mouse lung tumorigenesis. PMID- 12538358 TI - Acceleration of spontaneous biliary carcinogenesis in hamsters by bilioenterostomy. AB - Biliary carcinomas can occur as a delayed complication of bilioenterostomy. The aim of this study was to determine whether bilioenterostomy influences biliary carcinogenesis in hamsters. Syrian hamsters were subjected to three different surgical procedures: simple laparotomy (SL), choledochoduodenostomy (CD) and choledochojejunostomy (CJ). They were given no carcinogens, and five to six hamsters from each group were killed every 20 weeks up to 120 weeks after surgery. Thirty-seven, 32 and 38 hamsters were sampled from the SL, CD and CJ groups, respectively. Cholangiocarcinomas developed in 5.4, 15.6 and 23.7% of hamsters in the SL, CD and CJ groups, respectively. The incidence of biliary carcinoma was significantly higher in the bilioenterostomy groups, especially CJ (P < 0.05), than in SL. The tumor latency period after surgery was 20-40 weeks shorter in the bilioenterostomy groups than in SL. Persistent cholangitis and bile stasis were frequent in the bilioenterostomy groups, and a significant correlation between cholangitis and biliary carcinogenesis was noted in the CD group. The proliferative cell nuclear antigen (PCNA) labeling index was higher in the biliary epithelium of the bilioenterostomy groups. In conclusion, persistent cholangitis after bilioenterostomy accelerates biliary carcinogenesis through activation of biliary epithelial cell kinetics. PMID- 12538359 TI - Chemical carcinogens induce varying patterns of LOH in mouse T-lymphocytes. AB - We have shown previously that a wide range of mutagenic carcinogens are capable of inducing loss of heterozygosity (LOH) at the endogenous Aprt locus in mouse splenic lymphocytes. To investigate whether LOH might be caused by a single common mechanism, we set out to determine the extent of LOH by microsatellite analysis along (the Aprt gene containing) mouse chromosome 8. Aprt+/- hybrid B6C3F1 mice were treated with mutagens that induce different classes of DNA lesions, i.e. bulky DNA adducts, DNA methylation, DNA inter-strand crosslinks or DNA strand breaks. Aprt mutant frequencies (MF) in this C57Bl/6-C3H hybrid background were significantly reduced for mitomycin C (MMC) and methylmethanesulfonate (MMS) in comparison with MF in C57Bl/6 background, suggesting either enhanced repair or reduced formation of MMC- or MMS-induced mutagenic lesions in a hybrid B6C3F1 background. In contrast, Aprt MF after dimethylbenz[a]anthracene (DMBA), methylnitrosurea (MNU) and etoposide treatment were similar in both genetic backgrounds. Microsatellite analysis of Aprt mutant clones indicated a dominant role for mitotic recombination (MR) in generating spontaneous, DMBA- and etoposide-induced LOH at APRT: However, over 80% of the MMC-induced Aprt LOH mutants had lost heterozygosity for all markers tested, suggesting that either the crossover points were located close to the centromere or that these mutants arose by chromosome loss and duplication of the remaining chromosome 8. A substantial fraction (40%) of MNU-induced Aprt mutants had lost the wild-type Aprt allele, but had retained heterozygosity at all polymorphic markers tested at chromosome 8 indicating an important role for deletions in LOH formation by MNU. Patterns of MR differed quite dramatically for the various chemical mutagens tested, suggesting different mechanisms to be involved in inducing recombination between homologous chromosomes. In addition, non-random adduct formation and repair between chromosomal regions, i.e. heterochromatin versus euchromatin, may contribute to a non-random distribution of recombinational crossover points. PMID- 12538360 TI - CagA status of Helicobacter pylori infection and p53 gene mutations in gastric adenocarcinoma. PMID- 12538363 TI - Of mice and men (and rats): implications of species and stimulus differences for the interpretation of studies of nitric oxide in sepsis. PMID- 12538364 TI - Which anaesthetic agent for maintenance during normothermic cardiopulmonary bypass? PMID- 12538365 TI - Plasma propofol concentration and EEG burst suppression ratio during normothermic cardiopulmonary bypass. AB - BACKGROUND: During cardiopulmonary bypass (CPB), several factors affect drug disposition and action. This topic has not been studied extensively during normothermic CPB. In this study, we related propofol dose to plasma propofol concentration and burst suppression of the EEG during normothermic bypass. METHODS: After institutional approval and informed consent, 45 patients having cardiac surgery were assigned randomly to receive propofol infusions at 4 (Group A), 5 (Group B) and 6 (Group C) mg kg(-1) h(-1) during normothermic CPB. In all patients, small to moderate doses of fentanyl were also administered. Plasma propofol concentration and burst suppression ratio (BSR) were measured at the following times: (1) 10 min before CPB, (2) 10 min after the start of CPB, (3) 30 min after the start of the CPB, (4) just after aortic declamping, and (5) 60 min after CPB. RESULTS: At baseline, plasma propofol concentrations were similar among the three groups. After the start of CPB, the concentrations of propofol decreased significantly by 41, 35, and 30% of control values in Groups A, B, and C, respectively. In Group A, the concentration of propofol during CPB remained unchanged at less than the concentration before bypass. In Groups B and C, plasma propofol concentrations gradually increased during CPB to the pre-bypass concentrations. In Group A, BSR values did not change significantly during CPB. In Groups B and C, BSR values gradually increased and became significantly greater than baseline values. No patient reported intraoperative awareness. CONCLUSION: The pharmacokinetics and pharmacodynamics of propofol change during normothermic CPB. During normothermic CPB, the efficacy of propofol may be enhanced compared with before CPB. PMID- 12538366 TI - Relationship between bispectral index, auditory evoked potential index and effect site EC50 for propofol at two clinical end-points. AB - BACKGROUND: Many anaesthetists are deterred from using total i.v. anaesthesia because of uncertainty over the concentration of propofol required to prevent awareness. We predicted blood and effect-site concentrations of propofol at two clinical end-points: loss of consciousness and no response to a painful stimulus. METHODS: Forty unpremedicated Caucasian patients were anaesthetized with i.v. propofol delivered by a Diprifusor target-controlled infusion (TCI). Bispectral index (BIS) and auditory evoked potential index (AEPex) were measured and blood and effect-site propofol concentrations were predicted. Logistic regression was used to estimate population values for predicted blood and effect-site propofol concentrations at the clinical end-points and to correlate these with BIS and AEPex. RESULTS: The effect-site EC(50) at loss of consciousness was 2.8 micro m ml(-1) with an EC(05) and an EC(95) of 1.5 and 4.1 micro m ml(-1), respectively. The predicted EC(50) when there was no response to a tetanic stimulus was 5.2 micro m ml(-1) with an EC(05) and an EC(95) of 3.1 and 7.2 micro m ml(-1), respectively. CONCLUSIONS: Unconsciousness and lack of response to a painful stimulus occur within a defined range of effect-site concentrations, predicted by Diprifusor TCI software. PMID- 12538367 TI - Predictive performance of computer-controlled infusion of remifentanil during propofol/remifentanil anaesthesia. AB - BACKGROUND: The predictive performance of the available pharmacokinetic parameter sets for remifentanil, when used for target-controlled infusion (TCI) during total i.v. anaesthesia, has not been determined in a clinical setting. We studied the predictive performance of five parameter sets of remifentanil when used for TCI of remifentanil during propofol anaesthesia in surgical patients. METHODS: Remifentanil concentration-time data that had been collected during a previous pharmacodynamic interaction study in 30 female patients (ASA physical status I, aged 20-65 yr) who received a TCI of remifentanil and propofol during lower abdominal surgery were used in this evaluation. The remifentanil concentrations predicted by the five parameter sets were calculated on the basis of the TCI device record of the infusion rate-time profile that had actually been administered to each individual. The individual and pooled bias [median performance error (MDPE)], inaccuracy [median absolute performance error (MDAPE)], divergence and wobble of the remifentanil TCI device were determined from the pooled and intrasubject performance errors. RESULTS: A total of 444 remifentanil blood samples were analysed. Blood propofol and remifentanil concentrations ranged from 0.5 to 11 micro g ml(-1) and 0.1 to 19.6 ng ml(-1) respectively. Pooled MDPE and MDAPE of the remifentanil TCI device were -15 and 20% for the parameter set of Minto and colleagues (Anesthesiology 1997; 86: 10 23), 1 and 21%, -6 and 21%, and -6 and 19% for the three parameter sets described by Egan and colleagues (Anesthesiology 1996; 84: 821-33, Anesthesiology 1993; 79: 881-92, Anesthesiology 1998; 89: 562-73), and -24 and 30% for the parameter set described by Drover and Lemmens (Anesthesiology 1998; 89: 869-77). CONCLUSIONS: Remifentanil can be administered by TCI with acceptable bias and inaccuracy. The three pharmacokinetic parameter sets described by Egan and colleagues resulted in the least bias and best accuracy. PMID- 12538368 TI - Dynamic QRS-complex and ST-segment monitoring by continuous vectorcardiography during carotid endarterectomy. AB - BACKGROUND: Many authors report a high incidence of cardiac events during carotid endarterectomy. The aim of the present study was to evaluate the usefulness of dynamic continuous on-line vectorcardiography for monitoring the occurrence of myocardial ischaemia during carotid endarterectomy. METHODS: We studied 21 patients undergoing carotid endarterectomy. Patients underwent general anaesthesia with isoflurane or sevoflurane. The vectorcardiogram was monitored continuously during carotid endarterectomy. Electrodes were placed according to the previously described lead system and connected to a computerized system for on-line vectorcardiography. Two trend variables were recorded: the QRS vector difference, which reflects changes in the shape of the QRS complex; and the ST vector magnitude, which represents deflection of the ST segment from the isoelectric level. The ST segment deflection was measured 60 ms after termination of the QRS complex. RESULTS: Vectorcardiography was successfully recorded in all 21 patients. Three patients showed intraoperative vectorcardiogram abnormalities. In one of these three patients, both ST vector magnitude and QRS vector difference increased after induction of anaesthesia and ST vector magnitude returned to baseline after administration of nitroglycerin. In the other two patients, both ST vector magnitude and QRS vector difference gradually increased after cross-clamping of the internal carotid artery and ST vector magnitude returned to baseline after unclamping. QRS vector difference remained elevated for several hours in all three patients. CONCLUSIONS: Monitoring ST vector magnitude and QRS vector difference by vectorcardiography may be useful for identifying myocardial ischaemia during carotid endarterectomy. PMID- 12538370 TI - Pharmacokinetics and haemodynamics of ketamine in intensive care patients with brain or spinal cord injury. AB - BACKGROUND: Ketamine is used as an anaesthetic agent for short surgical procedures, and as a sedative and analgesic in intensive care patients. Intensive care patients with brain or spinal cord injury may have physiological changes that could alter the pharmacokinetics of ketamine. The pharmacokinetics of ketamine have been studied in healthy volunteers and in patients undergoing different types of surgery, but no data are available in intensive care patients. METHODS: We determined the pharmacokinetics of ketamine and its active metabolites, norketamine and dehydronorketamine, in 12 intensive care patients with brain or spinal cord injury. The effect of ketamine on haemodynamic variables was also investigated. RESULTS: The total clearance of ketamine, mean (SD), was 36.0 (13.3) ml min(-1) kg(-1), the volume of distribution (Vbeta) was 16.0 (8.6) litre kg(-1), and the elimination half-life was 4.9 (1.6) h. Ketamine did not alter any haemodynamic variables in the patients studied. CONCLUSIONS: Pharmacokinetic variables of ketamine in intensive care patients are greater than in healthy volunteers and in surgical patients. The increase in the volume of distribution is greater than the increase in clearance, resulting in a longer estimated half-life of ketamine in this patient group. PMID- 12538369 TI - Influence of intravenous vitamin E supplementation in cardiac surgery on oxidative stress: a double-blinded, randomized, controlled study. AB - BACKGROUND: I.V. infusions of vitamin E emulsion (all-rac-alpha-tocopherol) may reduce ischaemia-reperfusion injury after elective cardiac surgery. METHODS: Forty patients participated in a prospective, double-blind, placebo-controlled, randomized trial, receiving either placebo or four doses (270 mg each) of all-rac alpha-tocopherol between 16 h before and 48 h after surgery. We determined plasma concentrations of vitamin E, vitamin C, malondialdehyde, creatine kinase, troponin I and interleukin 6 and other measures of clinical outcome. RESULTS: Infusion of vitamin E caused normalization of vitamin E plasma concentrations during and after surgery, but had no effect on the early increase in malondialdehyde concentration or the decreases in antioxidative capacity and the water-soluble antioxidant vitamin C. CONCLUSIONS: Normalization of plasma vitamin E concentrations with parenteral vitamin E emulsion does not affect biochemical markers of myocardial injury and does not affect clinical outcome after cardiac surgery. PMID- 12538371 TI - Anaesthesia for awake craniotomy--evolution of a technique that facilitates awake neurological testing. AB - BACKGROUND: There is an increasing trend towards performing craniotomy awake. The challenge for the anaesthetist is to provide adequate analgesia and sedation, haemodynamic stability, and a safe airway, with an awake, cooperative patient for neurological testing. METHODS: The records of all patients who had awake craniotomy at our institution were reviewed. Patients were divided into three groups according to anaesthetic technique. Patients in Group 1 were sedated throughout the procedure. Patients in Groups 2 and 3 had an asleep-awake-asleep technique. Those in Group 2 were anaesthetized with a propofol infusion and fentanyl, and breathed spontaneously through a laryngeal mask airway (LMA). Patients in Group 3 had total i.v. anaesthesia with propofol and remifentanil, and ventilation was controlled using an LMA. We noted the incidence of complications in each group. RESULTS: There were 99 procedures carried out between 1989 and 2002. Group 3 had the fewest complications. No patients in Group 3 developed hypercapnia (E'(CO(2)) >6 kPa), compared with all of the patients in Group 2. Patients in Group 1 had no E'(CO(2)) monitoring, but 7% developed airway obstruction. No patients in Group 3 required additional analgesia for pain, compared with 70% of patients in Group 2. CONCLUSIONS: We have developed a technique for craniotomy, which facilitates awake neurological testing, is safe, and has good patient satisfaction. PMID- 12538372 TI - Parecoxib sodium has opioid-sparing effects in patients undergoing total knee arthroplasty under spinal anaesthesia. AB - BACKGROUND: This multicentre, double-blind, placebo-controlled study compared the opioid-sparing effectiveness and clinical safety of parecoxib sodium over 48 h, in 195 postoperative patients after routine total knee replacement surgery. METHODS: Elective total primary knee arthroplasty was performed under spinal anaesthesia, with a single dose of spinal bupivacaine 10-20 mg, and intraoperative sedation with midazolam 0.5-1.0 mg i.v., or propofol <6 mg kg( 1)h(-1). Patients were randomized to receive either parecoxib sodium 20 mg twice daily (bd) i.v. (n=65), parecoxib sodium 40 mg bd i.v. (n=67), or placebo (n=63) at the completion of surgery, and after 12, 24, and 36 h. Morphine (1-2 mg) was taken by patient-controlled analgesia or by bolus doses after 30 min. RESULTS: Patients receiving parecoxib sodium 20 mg bd and 40 mg bd consumed 15.6% and 27.8% less morphine at 24 h than patients taking placebo (both P<0.05). Both doses of parecoxib sodium administered with morphine provided significantly greater pain relief than morphine alone from 6 h (P<0.05). A global evaluation of study medication demonstrated a greater level of satisfaction among patients taking parecoxib sodium than those taking placebo. Parecoxib sodium administered in combination with morphine was well tolerated. However, a reduction in opioid type side-effects was not demonstrated in the parecoxib sodium groups. CONCLUSION: Parecoxib sodium provides opioid-sparing analgesic effects in postoperative patients. PMID- 12538373 TI - Block of the sacral segments in lumbar epidural anaesthesia. AB - BACKGROUND: Block of the first sacral segment is often delayed in lumbar epidural anaesthesia. The addition of either epinephrine or sodium bicarbonate to the local anaesthetic enhances the efficacy of epidural block. We assessed the block of lumbo-sacral segments in lumbar epidural anaesthesia adding epinephrine and/or bicarbonate to lidocaine. METHODS: Twenty-seven patients undergoing lumbar epidural anaesthesia with lidocaine 2%, 17 ml at L4-5 or L5-S1 were randomly divided into three groups. Plain lidocaine, lidocaine with 1:200,000 epinephrine or lidocaine-epinephrine-bicarbonate was administrated via an epidural catheter. The pain threshold after repeated electrical stimulation was used to assess the sensory block at the L2, S1, and S3 segments. Motor block was evaluated using the Bromage scale. RESULTS: Patient characteristics were comparable between the groups. The pH of lidocaine in the lidocaine-epinephrine-bicarbonate group was significantly higher than that in other groups. Pain thresholds at the S1 and S3 segments in the lidocaine-epinephrine-bicarbonate group were significantly higher than those in the lidocaine-epinephrine group. However, differences in the pain threshold at the L2 segment between groups were insignificant. The time to onset of sensory block at the S1 and S3 in the lidocaine-epinephrine-bicarbonate group was significantly shorter than that in the lidocaine group. Pain threshold by pinprick test was approximately within the 30-50 mA range. CONCLUSION: A combination of lidocaine, bicarbonate, and epinephrine increases the pain threshold over the sacral segments. PMID- 12538374 TI - Premixed solutions of diamorphine in ropivacaine for epidural anaesthesia: a study on their long-term stability. AB - BACKGROUND: Local anaesthetics and opioid mixtures are commonly used to provide anaesthesia or analgesia during the perioperative period. In order to facilitate their preparation and storage it is necessary to establish the stability of such solutions. METHODS: In our study, diamorphine was added to ropivacaine 0.2% 200 ml polybags to give a concentration of 25 micro g ml(-1) and to ropivacaine 1% 50 ml syringes to give a concentration of 45 micro g ml(-1). The polybags and syringes were stored at 40 degrees C, 21 degrees C and 4 degrees C for up to 120 days. Samples were taken during this period for measurement of diamorphine and ropivacaine content and pH of the solutions. RESULTS: We found that the storage temperature and the initial concentration influenced the rate of degradation of diamorphine in both the polybags and the syringes. In the syringes, 10% degradation of diamorphine [T (0.9)] was: 6 days at 40 degrees C, 16 days at 21 degrees C and 30 days at 4 degrees C. In the polybags, diamorphine T (0.9) was 6 days at 40 degrees C, 28 days at 21 degrees C and 70 days at 4 degrees C. CONCLUSIONS: It is feasible to manufacture such solutions in pharmacy aseptic units and to store them for up to 1 month for routine use in epidural infusions. PMID- 12538375 TI - Target controlled infusion of rocuronium: analysis of effect data to select a pharmacokinetic model. AB - BACKGROUND: We aimed to evaluate whether area under the curve (AUC) analysis of pharmacodynamic data can be used to compare pharmacokinetic models taken from the literature, during a target controlled infusion (TCI) of rocuronium. METHODS: Seventy-two patients scheduled for orthopaedic surgery received a TCI of rocuronium (Stanpump) based on one of four pharmacokinetic models: those described by Szenohradszky, Alvarez-Gomez, Wierda, and Cooper. The resulting theoretical plasma concentration versus time curve was calculated for all patients based on all four pharmacokinetic models. Predicted effect versus time curves were calculated following the pharmacokinetic-pharmacodynamic link model (Sheiner and colleagues). Neuromuscular block was evaluated acceleromyographically. The difference between the area under the observed effect (AUC(OE)) and predicted effect (AUC(PE)) versus time curves was used for comparison. RESULTS: AUC(PE )differed significantly from AUC(OE) in the Szenohradszky and Alvarez-Gomez models, both with the reference link pharmacodynamic data and with altered link-pharmacodynamic variables. AUC(PE )and AUC(OE) were comparable for the Wierda and Cooper models. The mean AUC(OE) was 25.1 (SD 11.9)% block x h. AUC(PE)-AUC(OE) was significantly larger in the Szenohradszky model when compared with all other pharmacokinetic models. This difference remained when link or pharmacodynamic variables were modified. The smallest AUC(PE)-AUC(OE) difference was found with the Wierda model. CONCLUSION: It was possible to use AUC analysis for identification of the pharmacokinetic model that best predicted the pharmacodynamic characteristics of our patients. PMID- 12538376 TI - Anaesthesia-related diplopia after cataract surgery. AB - BACKGROUND: We studied the incidence and clinical characteristics of persistent diplopia related to anaesthesia for cataract surgery in a general hospital. METHODS: This was a retrospective review of anaesthesia for 3587 cataract surgeries. Of all the cases of diplopia referred to the ocular motility clinic after cataract surgery, those involving anaesthesia-related diplopia lasting longer than 1 month were studied. RESULTS: During the study period, 3450 cataract surgeries were performed by phacoemulsification and 137 by extracapsular extraction. Retrobulbar block was used in 2024 cases, peribulbar block in 98, topical anaesthesia in 1420 and general anaesthesia in 43. Twenty-six cases of persistent diplopia were found (0.72% incidence), nine of which (0.25%) were considered to be related to anaesthetic factors; five of the latter involved the left eye. Five were caused by paresis of the inferior rectus muscle and three by fibrosis. In one patient, the inferior oblique muscle was affected. Anaesthesia was by retrobulbar block in eight cases (0.39%) and by peribulbar block in one. No diplopia was found in patients who had topical or general anaesthesia. Treatment was with surgery in two patients and with prisms in six. One patient continues to be studied. CONCLUSIONS: Persistent diplopia can occur after cataract surgery using retrobulbar block predominantly through direct damage to the inferior rectus muscle. The overall incidence of anaesthesia-related diplopia in this series was 0.25%. PMID- 12538377 TI - Dolasetron prophylaxis reduces nausea and postanaesthesia recovery time after remifentanil infusion during monitored anaesthesia care for extracorporeal shock wave lithotripsy. AB - BACKGROUND: Remifentanil is used as an analgesic for different procedures performed during monitored anaesthesia care. Opioid-induced nausea and vomiting can be troublesome. METHODS: This prospective, randomized, double-blind study was performed to evaluate the efficacy of prophylaxis with dolasetron in reducing the frequency of postoperative nausea and duration of discharge time. Forty urological patients, undergoing elective ambulatory extracorporeal shock wave lithotripsy (ESWL) received randomly either dolasetron 12.5 mg i.v. (Group 1) or placebo (Group 2) 10 min before a patient-adapted continuous infusion of remifentanil 0.15-0.4 micro g kg(-1) min(-1) was administered. Frequency and intensity (VAS 0-100 mm) of nausea, retching, and vomiting were assessed by patients and blinded investigators during and after the procedure. RESULTS: Patient characteristics, baseline values, duration of ESWL, and total dose of remifentanil did not differ between groups. The frequency (Group 1/Group 2; 20/55%; P<0.05) and mean (SD) maximal intensity [15 (9)/45 (14) mm; P<0.05] of nausea during 24 h was significantly reduced after dolasetron and discharge times in Group 1 were less than Group 2 [22 (14)/45 (28) min; P<0.05]. PMID- 12538378 TI - Differential interaction of anaesthetics and antiepileptic drugs with neuronal Na+ channels, Ca2+ channels, and GABA(A) receptors. AB - BACKGROUND: Current theories favour multiple agent-specific neuronal actions for both general anaesthetics and antiepileptic drugs, but the pharmacological properties that distinguish them are poorly understood. We compared the interactions of representative agents from each class on their putative targets using well-characterized radioligand binding assays. METHODS: Synaptosomes or membranes prepared from rat cerebral cortex were used to analyse drug effects on [(35)S]t-butyl bicyclophosphorothionate ([(35)S]TBPS) binding to the picrotoxinin site of GABA(A) receptors, [(3)H]batrachotoxinin A 20-alpha benzoate ([(3)H]BTX B) binding to site 2 of voltage-gated Na(+) channels, (+)-[methyl-(3)H]isopropyl 4-(2,1,3-benzoxadiazol-4-yl)-1,4-dihydro-5-methoxycarboxyl-2,6-dimethyl-3 pyridinecarboxylate ([(3)H]PN200-110; isradipine) binding to L-type Ca(2+) channels, and [cyclohexyl-2,3-(3)H](N)glibenclamide ([(3)H]GB) binding to K(ATP) channels. RESULTS: I.V. anaesthetics other than ketamine preferentially inhibited [(35)S]TBPS binding (etomidate approximately equal alphaxalone > propofol > thiopental > pentobarbital). Volatile anaesthetics inhibited both [(35)S]TBPS and [(3)H]BTX-B binding with comparable potencies (halothane approximately equal isoflurane approximately equal enflurane). Antiepileptic drugs preferentially antagonized either [(35)S]TBPS (diazepam > phenobarbital) or [(3)H]BTX-B (phenytoin > carbamazepine) binding. Local anaesthetics (lidocaine, tertracaine) selectively antagonized [(3)H]BTX-B binding. None of the drugs tested were potent antagonists of [(3)H]PN200-110 or [(3)H]GB binding. CONCLUSIONS: Comparative radioligand binding assays identified distinct classes of general anaesthetic and antiepileptic drugs based on their relative specificities for a defined target set. I.V. anaesthetics interacted preferentially with GABA(A) receptors, while volatile anaesthetics were essentially equipotent at Na(+) channels and GABA(A) receptors. Antiepileptic drugs could be classified by preferential actions at either Na(+) channels or GABA(A) receptors. Anaesthetics and antiepileptic drugs have agent-specific effects on radioligand binding. Both general anaesthetics and antiepileptic drugs interact with Na(+) channels and GABA(A) receptors at therapeutic concentrations, in most cases with little selectivity. PMID- 12538379 TI - Effects of epidural anaesthesia on intestinal oxygenation in pigs. AB - BACKGROUND: Perioperative intestinal hypoperfusion is a major contributing factor leading to organ dysfunction. It can be caused by stress as a result of surgical manipulation or hypoxia. Additionally, anaesthesia can affect intestinal oxygenation. This animal study was designed to assess the effects of reduced regional sympathetic nervous activity induced by thoracic epidural anaesthesia on intestinal oxygenation. METHODS: After ethical approval, 16 anaesthetized and acutely instrumented pigs were randomly assigned to two groups (epidural anaesthesia alone vs epidural anaesthesia plus volume loading). The epidural anaesthesia aimed for a T5-T12 block. Measurements were at baseline and after 1 and 2 h. RESULTS: Epidural anaesthesia was associated with a decrease in mean arterial blood pressure and pronounced mesenteric vasodilatation. Mesenteric blood flow did not change. Intestinal oxygen uptake, mucosal tissue oxygen partial pressure and tissue carbon dioxide partial pressure remained unchanged. CONCLUSIONS: Despite marked systemic hypotension, epidural anaesthesia did not affect intestinal oxygenation. There was no benefit obtained from volume loading. PMID- 12538380 TI - Oxidative stress and gene expression in sepsis. AB - Dysregulation of the immuno-inflammatory response, as seen in sepsis, may culminate in host cell and organ damage. Lipopolysaccharide from Gram-negative bacterial cell walls induces gene activation and subsequent inflammatory mediator expression. Gene activation is regulated by a number of transcription factors at the nuclear level, of which nuclear factor kappaB appears to have a central role. The redox (reduction-oxidation) cellular balance is important for normal cellular function, including transcription factor regulation. In sepsis, a state of severe oxidative stress is encountered, with host endogenous antioxidant defences overcome. This has implications for cellular function and the regulation of gene expression. This review gives an overview of the mechanisms by which transcription factor activation and inflammatory mediator overexpression occur in sepsis, together with the events surrounding the state of oxidative stress encountered and the effects on the host's antioxidant defences. The effect of oxidative stress on transcription factor regulation is considered, together with the role of antioxidant repletion in transcription factor activation and in sepsis in general. Other interventions that may modulate transcription factor activation are also highlighted. PMID- 12538381 TI - Sevoflurane anaesthesia does not induce the formation of sister chromatid exchanges in peripheral blood lymphocytes of children. AB - BACKGROUND: Compound A, a degradation product of sevoflurane, has been demonstrated to induce sister chromatid exchanges (SCE) in Chinese hamster ovary cells in vitro as a marker for possible genotoxicity. We investigated the formation of SCE in mitogen-stimulated T-lymphocytes of 40 children undergoing sevoflurane anaesthesia for minor surgical procedures. METHODS: Anaesthesia was induced by inhalation of up to 8% sevoflurane and maintained at 2.5-3% in oxygen/nitrous oxide (65/35%) at a fresh gas flow of 3 litre min(-1). Soda lime (humidity 12-15%) was used as a carbon dioxide absorbent. Blood was drawn directly before induction and after termination of anaesthesia. Twenty-five second division metaphases of mitogen-stimulated T-lymphocytes per blood sample were screened for SCE rates using standard techniques. RESULTS: Average duration of anaesthesia was 49.6 (SD 24.0) min. Before anaesthesia induction, 7.93 (1.23) SCE per metaphase were determined. After sevoflurane anaesthesia [1.40 (0.77) MAC h] 7.92 (1.19) SCE per metaphase were observed. Additionally, no differences were evident between male or female children. CONCLUSION: Short-term administration of sevoflurane anaesthesia did not induce SCE in T-lymphocytes of children. No indication for a possible genotoxic effect has been observed. PMID- 12538382 TI - Depth of insertion of the ProSeal laryngeal mask airway. AB - BACKGROUND: The depth of insertion of the ProSeal laryngeal mask airway (PLMA) is unknown. We measured depth of insertion in satisfactorily positioned PLMAs. METHODS: All women received size 4 masks and men size 5 masks. We measured the position of the integral bite block in relation to the upper incisors documented in patients over a 6-month period. Depth of insertion was scored by dividing the integral bite block into quarters. Satisfactory positioning of the ProSeal itself was determined by (i) positive 'suprasternal notch test', (ii) no venting via the drain tube during maximal lung inflation, and (iii) an unobstructed airway. RESULTS: We studied 274 patients (147 women and 127 men). The midway point of the bite block was proximal to the incisors (e.g. within the oropharynx) in 78% of women (95% CI 71-85%) and 92% of men (95% CI 87-97). The standard deviation for the depth distribution in women was 0.8 cm and for men was 1.0 cm. CONCLUSIONS: Usually most of the integral bite block lies within the oropharynx. It was never normal for the entire bite block to stick out of the mouth (4 SD from the mean for both men and women). The position of the integral bite block relative to the upper incisors gives valuable information during assessment of PLMA position. PMID- 12538383 TI - Use of prone ventilation in neurogenic pulmonary oedema. AB - We present a case of neurogenic pulmonary oedema (NPO) due to subarachnoid haemorrhage that resulted in hypoxia refractory to conventional mechanical ventilation. Prone positioning was employed, resulting in rapid and sustained improvement in oxygenation. We discuss the pathogenesis of NPO and the mechanism of action of prone ventilation. Prone ventilation may be of value in the management of NPO, both in treating life-threatening hypoxia and in optimizing neurological recovery. Further data are required on its effect on intracranial pressure after subarachnoid haemorrhage. PMID- 12538384 TI - Anaesthetic management of a parturient with pulmonary stenosis and aortic incompetence for Caesarean section. AB - Anaesthetic management of Caesarean section in a parturient with severe pulmonary stenosis and aortic regurgitation is described. The valvular sequelae resulted from previous unsuccessful surgical correction (Ross procedure) of congenital aortic stenosis. This case demonstrates the importance of multi-disciplinary assessment and careful anaesthetic planning, to avoid deterioration in perioperative cardiac performance in parturients with complex valvular disease. PMID- 12538385 TI - Non-fatal amniotic fluid embolism after cervical suture removal. AB - We describe a case of pulmonary oedema occurring at 37 weeks gestation, following the attempted removal of a cervical suture under general anaesthesia. The use of an ultrasound technique to demonstrate the patient's fluid status is described. Signs of amniotic fluid embolism and how it exerts its influence on the circulation are discussed. PMID- 12538386 TI - Recurrent post-partum seizures after epidural blood patch. AB - There are many causes for headaches after childbirth. Even though postdural puncture headache (PDPH) has to be considered in a woman with a history of difficult epidural anaesthesia, pre-eclampsia should always be excluded as an important differential diagnosis. We report a case with signs of late-onset pre eclampsia where administration of an epidural blood patch (EBP) was associated with eclampsia. A hypothetical causal relationship between the EBP and seizures was discarded on the basis of evidence presented in this report. PMID- 12538387 TI - Effect of hypothermia on brain tissue oxygenation in patients with severe head injury. PMID- 12538388 TI - Evaluation of platelet aggregation in flow and platelet aggregometry during pregnancy. PMID- 12538389 TI - Propofol and myocardial lipid peroxidation. PMID- 12538390 TI - Comparison of morphine alone with morphine plus clonidine for postoperative patient-controlled analgesia. PMID- 12538391 TI - Continuous and intermittent cardiac output measurement: pulmonary artery catheter versus aortic transpulmonary technique. PMID- 12538392 TI - Confidential enquiries into maternal deaths, 1997-1999. PMID- 12538393 TI - Training course in local anaesthesia of the airway and fibreoptic intubation using course delegates as subjects. PMID- 12538394 TI - Venepuncture as an antiarrhythmic. PMID- 12538395 TI - Mind-the gap, after 65 years: visual conditioning in cortical blindness. PMID- 12538396 TI - Affective blindsight: intact fear conditioning to a visual cue in a cortically blind patient. AB - Blindsight refers to remarkable residual visual abilities of patients with damage to the primary visual cortex (V1). Recent studies revealed that such residual abilities do not apply only to relatively simple object discriminations, but that these patients can also differentially categorize and respond to emotionally salient stimuli. The current study reports on a case of intact fear conditioning to a visual cue in a male patient with complete bilateral cortical blindness. The patient was admitted to the stroke unit of the neurological department because of complete loss of vision. Both CT and structural MRI scans confirmed lesions in both territories of the posterior cerebral artery. No visual evoked potentials could be detected confirming complete cortical blindness. During fear conditioning, a visual cue predicted the occurrence of an aversive electric shock. Acoustic startle probes were presented during and between the conditioned stimuli. Relative to the control condition, startle reflexes were substantially potentiated when elicited in the presence of the conditioned stimuli. No such potentiation was observed prior to conditioning. These data suggest that fear learning to visual cues does not require a cortical representation of the conditioned stimulus in the primary sensory cortex and that subcortical pathways are sufficient to activate the fear module in humans. PMID- 12538397 TI - Chronic relapsing inflammatory optic neuropathy (CRION). AB - We describe the clinical characteristics and early natural history of a form of inflammatory optic neuropathy which is frequently bilateral and often painful, and is characterized by relapses and remissions. MRI scans of the brain are normal and those of the optic nerves often, but not always, show high signal abnormalities which enhance. The symptoms and signs respond well to corticosteroid treatment, although long-term immuno suppression is often necessary. The syndrome behaves in a way which is typical of the condition known as granulomatous optic neuropathy, but during a median follow-up of 8 (2-26) years in no case has evidence for systemic sarcoidosis been identified. We suggest that the disorder be named chronic relapsing inflammatory optic neuropathy (CRION). PMID- 12538398 TI - Vaccination with amyloid-beta peptide induces autoimmune encephalomyelitis in C57/BL6 mice. AB - Recent experimental evidence shows that vaccination with amyloid-beta peptide (Abeta) of transgenic mouse models of Alzheimer's disease protects from the pathological accumulation of amyloid within the CNS. Phase I/II clinical trials of Abeta vaccination in mild to moderate Alzheimer's disease have been undertaken. Un expectedly, one of these trials has been suspended because 15 patients showed clinical signs consistent with CNS inflammation. Here, we show that C57BL/6 mice immunized with Abeta1-42 peptide develop an inflammatory disease of the CNS characterized by the presence both in the brain and spinal cord of perivenular inflammatory foci containing macrophages, T and B cells, and immunoglobulins. The experimental disease was observed only when pertussis toxin, an agent known to favour autoimmune processes, was co-administered. The immune mediated CNS reaction was associated to Abeta-induced CD4(+) cells showing a Th1 type cytokine expression profile and to elevated levels of circulating anti-Abeta immunoglobulins. Our results indicate that vaccination with Abeta could determine, under certain circumstances, an aberrant autoimmune-type reaction to Abeta resulting in a perivenular inflammatory encephalomyelitis. PMID- 12538399 TI - Neural basis for semantic memory difficulty in Alzheimer's disease: an fMRI study. AB - Patients with probable Alzheimer's disease are thought to have a semantic memory deficit. We used functional MRI to evaluate the neural basis for impaired semantic memory for ANIMALS and IMPLEMENTS in 11 patients with Alzheimer's disease and 16 healthy seniors. For both categories of knowledge, Alzheimer's disease patients show reduced activation in the left posterolateral temporal inferior parietal cortex compared with healthy seniors. Activation changes in this heteromodal association region may be related to an impairment of the category-neutral semantic processes involved in integrating feature knowledge that is represented in modality-specific association cortices. We also found increased activation of an area of the left temporal cortex for both categories of knowledge in Alzheimer's disease that was not activated in healthy seniors. Category-specific changes were also seen in Alzheimer's disease compared with healthy seniors that may be related to the neural representation of category specific feature knowledge represented in semantic memory. For ANIMALS, the left ventral temporal cortex was activated in Alzheimer's disease in an anatomical distribution that was posterior to the left ventral recruitment for this category in healthy seniors. For IMPLEMENTS, frontal-striatal regions were activated in Alzheimer's disease in a manner that was displaced from the locus of recruitment for this category in healthy seniors. Our findings are consistent with a two component model of semantic memory involving category-neutral processes operating on category- specific knowledge, and both components appear to be compromised in Alzheimer's disease. Components of the large-scale neural network underlying semantic memory may modify themselves to maintain performance in the face of a neurodegenerative disease. PMID- 12538401 TI - Abnormal corticomuscular and intermuscular coupling in high-frequency rhythmic myoclonus. AB - Frequency analysis may have some advantages over back-averaging in the neurophysiological assessment of patients with suspected cortical myoclonus in whom myoclonic EMG bursts repeat rhythmically at high frequency. However, the clinical utility of EEG-EMG coherence and related EMG-EMG coherence is not established. Equally, there is an incomplete understanding of the physiology of the systems contributing to the coherence evident between signals in cortical myoclonus. Here we address these issues in an investigation of EEG-EMG and EMG EMG coupling in proximal and distal muscles of the upper extremities in nine patients with multifocal high frequency rhythmic myoclonus due to non-progressive conditions. We found exaggerated coherence between EEG and contralateral EMG and between pairs of ipsilateral EMG signals. The results of frequency analysis of EMG-EMG mirrored those for EEG-EMG, but the former technique was superior in distinguishing a pathologically exaggerated common drive in distal upper limb muscles. Both techniques were more sensitive than back-averaging. Frequency analysis also revealed important disparities between proximal and distal upper limb muscles. In the latter case, the functional coupling between cortex and muscle was dominated by efferent processes. In contrast, there was considerable inter-individual variation in the extent to which EEG-EMG and EMG-EMG coupling in proximal muscles reflected afferent and efferent loops. Thus, the processes sustaining myoclonic discharges may differ for proximal and distal muscles and between patients. PMID- 12538400 TI - Dopamine release during sequential finger movements in health and Parkinson's disease: a PET study. AB - Parkinson's disease is associated with slowness, especially of sequential movements, and is characterized pathologically by degeneration of dopaminergic neurons, particularly targeting nigrostriatal projections. In turn, nigrostriatal dopamine has been suggested to be critical for the execution of sequential movements. The objective of this study was to investigate in vivo, with [(11)C]raclopride, PET changes in regional brain levels of dopamine in healthy volunteers and Parkinson's disease patients during the execution of paced, stereotyped sequential finger movements. Striatal [(11)C]raclopride binding reflects dopamine D(2) receptor availability and is influenced by synaptic levels of endogenous dopamine. During execution of a pre-learned sequence of finger movements, a significant reduction in binding potential (BP) of [(11)C]raclopride was seen in both caudate and putamen in healthy volunteers compared with a resting baseline, consistent with release of endogenous dopamine. Parkinson's disease patients also showed attenuated [(11)C]raclopride BP reductions during the same motor paradigm in striatal areas less affected by the disease process. These findings confirm that striatal dopamine release is a component of movement sequencing and show that dopamine release can be detected in early Parkinson's disease during a behavioural manipulation. PMID- 12538402 TI - Pantomime to visual presentation of objects: left hand dyspraxia in patients with complete callosotomy. AB - Investigations of left hand praxis in imitation and object use in patients with callosal disconnection have yielded divergent results, inducing a debate between two theoretical positions. Whereas Liepmann suggested that the left hemisphere is motor dominant, others maintain that both hemispheres have equal motor competences and propose that left hand apraxia in patients with callosal disconnection is secondary to left hemispheric specialization for language or other task modalities. The present study aims to gain further insight into the motor competence of the right hemisphere by investigating pantomime of object use in split-brain patients. Three patients with complete callosotomy and, as control groups, five patients with partial callosotomy and nine healthy subjects were examined for their ability to pantomime object use to visual object presentation and demonstrate object manipulation. In each condition, 11 objects were presented to the subjects who pantomimed or demonstrated the object use with either hand. In addition, six object pairs were presented to test bimanual coordination. Two independent raters evaluated the videotaped movement demonstrations. While object use demonstrations were perfect in all three groups, the split-brain patients displayed apraxic errors only with their left hands in the pantomime condition. The movement analysis of concept and execution errors included the examination of ipsilateral versus contralateral motor control. As the right hand/left hemisphere performances demonstrated retrieval of the correct movement concepts, concept errors by the left hand were taken as evidence for right hemisphere control. Several types of execution errors reflected a lack of distal motor control indicating the use of ipsilateral pathways. While one split-brain patient controlled his left hand predominantly by ipsilateral pathways in the pantomime condition, the error profile in the other two split-brain patients suggested that the right hemisphere controlled their left hands. In the object use condition, in all three split-brain patients fine-graded distal movements in the left hand indicated right hemispheric control. Our data show left hand apraxia in split brain patients is not limited to verbal commands, but also occurs in pantomime to visual presentation of objects. As the demonstration with object in hand was unimpaired in either hand, both hemispheres must contain movement concepts for object use. However, the disconnected right hemisphere is impaired in retrieving the movement concept in response to visual object presentation, presumably because of a deficit in associating perceptual object representation with the movement concepts. PMID- 12538403 TI - Autoimmunoreactivity to Schwann cells in patients with inflammatory neuropathies. AB - Inflammatory demyelinating neuropathies are characterized by a loss of peripheral nerve myelin. Myelin breakdown is thought to result from an autoimmune reaction towards nerve components. Schwann cells play a crucial role in the synthesis and maintenance of peripheral nerve myelin. An immune attack targeting Schwann cells could therefore affect myelin integrity, leading to disease. We studied the reactivity of sera from patients with Guillain-Barre syndrome (GBS) and chronic inflammatory demyelinating polyneuropathy (CIDP) towards Schwann cells using immunofluorescence microscopy. We found 24% of the GBS (56 out of 233) and 26% of the CIDP (12 out of 46) patients to have circulating immunoglobulin G autoantibodies against proliferating, non-myelinating human Schwann cells. In contrast, healthy donors showed positive staining in only two out of 34 sera. No reaction was found with sera from patients with non-inflammatory neurological disorders. Immunofluorescence was localized at the distal tips (leading lamella) of the Schwann cell processes. Distal tips of neurites (nerve-growth-cones) of in vitro differentiated non-myelinated hNT2 neurons also stained strongly. GBS and CIDP serum immunoreactivity was also observed in teased nerve fibre preparations. These data suggest that, at least part of the immunoreactivity is not directed against myelin, but towards non-myelin proteins and epitopes possibly involved in Schwann cell-axon interaction. PMID- 12538404 TI - Inflammatory vasculopathy in multifocal diabetic neuropathy. AB - Besides the common distal symmetrical sensory-motor polyneuropathy (DSP) that is often associated with autonomic dysfunction, diabetic patients may develop multifocal sensory-motor deficits (MDN) secondary to roots, plexus and nerve trunk involvement. Nerve ischaemia has been suggested as a common mechanism for the different patterns of diabetic neuropathies, yet the important clinical differences that exist between DSP and MDN suggest concurrent factors. In order to learn more on the subject, we prospectively studied 22 consecutive diabetic patients with MDN, for which other causes of neuropathy were excluded by appropriate investigations, including biopsy of a recently affected sensory nerve. Three patients had a relapsing course, and the others an unremitting subacute-progressive course. Painful MDN progressed over 2-12 months. The neurological deficit predominated in distal lower limbs which were involved in all patients, unilaterally in seven, bilaterally in the others, with an asynchronous onset in most cases. In addition, a proximal deficit of the lower limbs was present on one side in seven patients, and on both sides in six. Thoracic radiculoneuropathy was present bilaterally in two patients, and unilaterally in one. The ulnar nerve was involved in one patient, and the radial nerve in two. The CSF protein ranged from 0.40 to 3.55 g/l; mean: 0.87 g/l. Electrophysiological testing showed severe, multifocal, axonal nerve lesions in all cases. Asymmetrical axonal lesions were found in all nerve specimens. The mean density of myelinated axons was reduced to 1340 +/- 1070 per mm(2) of endoneurial area versus 8370 +/- 706 myelinated fibres/mm(2) in controls. The mean density of unmyelinated fibres was reduced to 5095 +/- 6875 per mm(2) (extremes: 0-26 600). On teased fibre preparations, 34 +/- 31% of the fibres were at different stages of axonal degeneration (extremes 0-99%); 7 +/- 6% of the fibres showed segmental demyelination or remyelination. Necrotizing vasculitis of perineurial and endoneurial blood vessels were found in six patients. Endoneurial seepage of red cells was present in 11 specimens, and endoneurial haemorrhage in five. Ferric iron deposits that characterize previous bleeding were found in seven patients, including two who had no red cells in the endoneurium. Perivascular mononuclear cell infiltrates were present in the nerve specimens of 21 out of 22 patients, prominently in four patients. In comparison, nerve biopsy specimens of 30 patients with severe distal symmetrical diabetic polyneuropathy showed mild epineurial mononuclear cell infiltrate in one patient and endoneurial seepage of red cells in another. We conclude that MDN is related to pre-capillary blood vessel involvement in elderly diabetic patients with a secondary inflammatory response. PMID- 12538405 TI - Cutaneous innervation in Guillain-Barre syndrome: pathology and clinical correlations. AB - Guillain-Barre syndrome (GBS) is traditionally considered to be a large-fibre neuropathy. However, the presence of hypo-aesthesia, dysaesthesia and dysautonomia in GBS patients raises the possibility that small-diameter sensory and autonomic nerves may also be affected. To investigate small-fibre neuropathy in GBS, we performed a skin biopsy from the distal leg of 20 patients with the demyelinating form of GBS. Skin sections were immunohistochemically stained with antiserum against protein gene product 9.5 (PGP 9.5), a ubiquitin C-terminal hydrolase. Cutaneous innervation was evaluated by measuring epidermal nerve density (END), and END was further correlated with various clinical and electrophysiological parameters. In GBS patients, END values were much lower than in age- and gender-matched control subjects (5.03 +/- 1.18 versus 10.16 +/- 0.87 fibres/mm, P < 0.001). Eleven patients (55%) had reduced epidermal innervation with pathological evidence of active nerve degeneration in the dermis: fragmentation of subepidermal nerve plexuses and a beaded appearance of dermal nerves. GBS patients had significantly elevated thermal thresholds with higher warm threshold temperatures (44.54 +/- 1.04 versus 39.00 +/- 0.35 degrees C, P < 0.001) and lower cold threshold temperatures (25.57 +/- 1.11 versus 29.05 +/- 0.21 degrees C, P = 0.032). Reduced END values were associated with an elevated warm threshold (P = 0.027), ventilatory distress (P = 0.037) and dysautonomia (P = 0.001). END values were negatively correlated with disability grade on a scale of 1-6 (slope -0.134 +/- 0.038, P = 0.0018). Patients with reduced END values tended to have a slower recovery than those with normal END values (P = 0.013, median time 12 versus 2 weeks). Patho logically, sudomotor innervation of the skin was reduced in five of 17 (29.4%) GBS patients in whom sweat glands could be recognized. These findings suggest that small-fibre sensory and autonomic neuropathies exist in a significant proportion of GBS patients, and that END values are correlated with functional disabilities. In summary, GBS should be considered a global neuropathy instead of a pure large-fibre neuropathy. PMID- 12538406 TI - Regulation of gene expression in experimental autoimmune encephalomyelitis indicates early neuronal dysfunction. AB - Multiple sclerosis is an inflammatory, demyelinating disease of the CNS. Whereas oligodendrocytes have been considered the primary neural cell type most affected, recent evidence indicates that axonal and neuronal degeneration also occurs in both multiple sclerosis and experimental autoimmune encephalomyelitis (EAE), an animal model reproducing many features of multiple sclerosis. The molecular mechanisms underlying neuronal deficits in multiple sclerosis and EAE remain elusive. To address this issue, we have analysed the expression of genes encoding proteins that play critical roles in ion homeostasis, exocytosis, mitochondrial function and impulse conduction in the Lewis rat lumbar spinal cord during the clinical course of acute EAE. Transcript and protein levels of plasma membrane Ca(2+) ATPase 2 (PMCA2), an essential ion pump expressed exclusively in grey matter and involved in Ca(2+) extrusion, synapsin IIa and syntaxin 1B, important regulators of vesicular exocytosis, were dramatically decreased coincident with the onset of clinical symptoms. In contrast, changes in the expression of several other ion pumps, vesicular proteins, mitochondrial enzymes and sodium channels occurred at more advanced disease stages. Moreover, exposure of spinal cord slice cultures to kainic acid significantly reduced PMCA2 mRNA levels. Taken together, our findings suggest that glutamate, which recently has been implicated in EAE pathogenesis, suppresses neuronal PMCA2 expression leading to Ca(2+) dyshomeostasis at initial clinical phases. Consequently, perturbations in Ca(2+) balance and neurotransmitter exocytosis may partially underlie aberrant neuronal function and communication at onset of symptoms. Altered mitochondrial function and impulse conduction may exacerbate neurological deficits at subsequent disease stages. PMID- 12538407 TI - The spectrum of exercise tolerance in mitochondrial myopathies: a study of 40 patients. AB - Impaired skeletal muscle oxidative phosphorylation in patients with severe mitochondrial respiratory chain defects results in disabling exercise intolerance that is associated with a markedly blunted capacity of muscle to increase oxygen utilization in relation to circulatory and ventilatory responses that increase oxygen delivery to muscle during exercise. The range of oxidative limitation and the relationship between the severity of oxidative defects and physiological responses to exercise among a broader spectrum of mitochondrial respiratory chain defects has not been defined. We evaluated oxidative capacity and circulatory and ventilatory responses to maximal cycle exercise in 40 patients with biochemically and/or molecularly defined mitochondrial myopathy (MM) associated with varying levels of exercise tolerance, and compared responses with those in healthy sedentary individuals. In the MM patients, mean peak work capacity (0.88 +/- 0.6 W/kg) and oxygen uptake (VO(2), 16 +/- 8 ml/kg/min) were significantly lower (P < 0.01) than in controls (mean work capacity = 2.2 +/- 0.7 W/kg; VO(2) = 32 +/- 7 ml/kg/min), but the patient range was broad (0.17-3.2 W/kg; 6-47 ml/kg/min). Oxidative capacity in patients was limited by the ability of muscle to extract available oxygen from blood [mean peak systemic arteriovenous O(2) difference (a vO(2)); patients = 7.7 +/- 3.5, range 2.7-17.6 ml/dl, controls = 15.2 +/- 2.1 ml/dl], as indicated by a linear correlation between peak VO(2) and peak systemic a-vO(2) difference (r(2) = 0.69). In the patients, the increase in cardiac output relative to VO(2) (mean DeltaQ/DeltaVO(2) = 15.0 +/- 13.6; range 3.3-73) and ventilation (mean peak VE/VO(2) = 65 +/- 24; range 21-104) were exaggerated compared with controls (mean DeltaQ/DeltaVO(2) = 5.1 +/- 0.7; VE/VO(2) = 41.2 +/- 7.4, P < 0.01). There was a negative exponential relationship between DeltaQ/DeltaVO(2) and peak systemic a-vO(2) difference (r(2) = 0.92) and between peak VE/VO(2) and systemic a-vO(2) difference (r(2) = 0.53). In patients with heteroplasmic mtDNA mutations, we found an inverse relationship between the proportion of skeletal muscle mutant mtDNA and peak extraction of available oxygen during exercise (r(2) = 0.70). We conclude that the degree of exercise intolerance in MM correlates directly with the severity of impaired muscle oxidative phosphorylation as indicated by the peak capacity for muscle oxygen extraction. Exaggerated circulatory and ventilatory responses to exercise are direct consequences of the level of impaired muscle oxidative phosphorylation and increase exponentially in relation to an increasing severity of oxidative impairment. In patients with mtDNA mutations, muscle mutation load governs mitochondrial capacity for oxidative phosphorylation and determines exercise capacity. PMID- 12538408 TI - Markers of endothelial dysfunction in lacunar infarction and ischaemic leukoaraiosis. AB - Patients with cerebral small vessel disease (SVD) can present as isolated lacunar infarction or with diffuse white matter changes, with the imaging appearance of leukoaraiosis. Endothelial dysfunction, which can lead to breakdown of the blood brain barrier, impaired cerebral autoregulation and prothrombotic changes, is believed to be important in mediating disease. Circulating levels of intercellular adhesion molecule 1 (ICAM1), thrombomodulin (TM), tissue factor (TF) and tissue factor pathway inhibitor (TFPI) are markers of endothelial activation and damage, and may provide insights into disease pathogenesis or differences between phenotypes. We therefore measured these markers in a prospective series of patients with lacunar stroke. One hundred and ten white Caucasian patients with previous lacunar stroke and 50 community control subjects were studied. Markers of endothelial function were measured on venous blood samples. Patients were classified on brain imaging into two groups: isolated lacunar infarction (n = 47) and ischaemic leukoaraiosis, defined as a clinical lacunar stroke and leukoaraiosis on brain imaging (n = 63). The number of lacunes and severity of leukoaraiosis were also scored on MRI. ICAM1, TM and TFPI were elevated in cerebral SVD subjects compared with controls (P 0.38 mg/dL) had inducible ischemia, compared with 45% (79/177) in the lower 4 categories combined (adjusted odds ratio 4.2; 95% confidence interval 1.6 to 11; P=0.004). However, this association differed in users and nonusers of beta-blockers and statins. Among 89 participants who did not use beta-blockers, 93% in the highest CRP category had exercise-induced ischemia, compared with 42% in the lower 4 categories (P=0.03). Among 67 participants who did not use statins, 94% in the highest CRP category had exercise-induced ischemia, compared with 44% in the lower 4 categories (P=0.009). We did not observe a significant association between CRP and ischemia among participants who were treated with either of these medications. CONCLUSION: Elevated CRP levels are associated with inducible ischemia in patients with stable coronary disease, particularly among those not treated with beta-blockers or statins. PMID- 12538424 TI - Multiple infections and subsequent cardiovascular events in the Heart Outcomes Prevention Evaluation (HOPE) Study. AB - BACKGROUND: Limited prospective epidemiological data are available on the relation between exposure to Chlamydia pneumoniae, Helicobacter pylori, cytomegalovirus (CMV), and hepatitis A virus (HAV), individually or as a total pathogen score, and human cardiovascular (CV) disease. METHODS AND RESULTS: We analyzed enrollment sera from 3168 Canadian patients in the Heart Outcomes Prevention Evaluation (HOPE) study for antibodies to C pneumoniae, H pylori, CMV, and HAV and measured the relation between serostatus and 494 adjudicated trial outcomes of myocardial infarction, stroke, or CV death over 4.5 years of follow up. CV events were associated with CMV serostatus (covariate-adjusted hazard ratio [HR], 1.24; 95% CI, 1.01, 1.53). Neither C pneumoniae IgG (adjusted HR, 0.87; 95% CI, 0.68, 1.10), C pneumonia IgA (adjusted HR, 1.10; 95% CI, 0.90, 1.34), H pylori IgG (HR, 0.99; 95% CI, 0.82, 1.19), nor HAV IgG (HR, 1.01; 95% CI, 0.83, 1.24) predicted CV events. Total pathogen score was associated with CV events (adjusted HR for 4 versus 1 or 0=1.41; 95% CI, 1.02, 1.96). CONCLUSIONS: Exposure to CMV but not to C pneumoniae, H pylori, or HAV was associated with a slight excess risk of subsequent myocardial infarction, stroke, or CV death in HOPE study patients, and total pathogen score based on these infections predicted a small increased hazard of CV events. PMID- 12538425 TI - Statin therapy interacts with cytomegalovirus seropositivity and high C-reactive protein in reducing mortality among patients with angiographically significant coronary disease. AB - BACKGROUND: Seropositivity to cytomegalovirus (CMV) and elevated C-reactive protein (CRP) may jointly predict increased mortality rates in patients with coronary artery disease (CAD). Therapy with statins reduces lipid levels but may also have other beneficial (eg, antiinflammatory) effects. This study prospectively evaluated the effect of statins on CMV-and CRP-associated death among patients with significant, angiographically defined CAD. METHODS AND RESULTS: We monitored 2315 patients with angiographically significant CAD (stenosis > or =70%) for an average of 2.4 years (maximum, 5.8 years). Anti-CMV IgG antibody levels and CRP concentrations were measured at baseline, and statin prescription was recorded. As previously reported, mortality rate was higher for CMV seropositivity (+) with high CRP (hazard ratio [HR], 2.0) and lower for statins (HR, 0.50). Compared with CMV(-)/low CRP (mortality rate, 5% with statin versus 4% without statin), the protective effect of statin therapy was markedly greater for CMV(+)/low CRP (mortality rate, 2% versus 7%; HR, 0.44; 95% CI, 0.16 to 1.3), CMV negative (-)/high CRP (mortality rate, 1% versus 8%; HR, 0.16), and CMV(+)/high CRP (mortality rate, 6% versus 17%; HR, 0.42; 95% CI, 0.25 to 0.70). After adjustment, interactions were found for statin therapy with CMV(+)/low CRP (P for interaction=0.065), CMV(-)/high CRP (P for interaction=0.051), and CMV(+)/high CRP (P for interaction=0.024). CONCLUSIONS: The survival benefit of statins interacted with CMV seropositivity and high CRP to significantly reduce mortality rates among patients with CAD. This finding supports the hypothesis that statins have beneficial, "lipid-independent," antiinflammatory effects. The mechanism of statin benefit associated with CMV seropositivity remains to be determined. PMID- 12538426 TI - Serum from patients with acute coronary syndromes displays a proapoptotic effect on human endothelial cells: a possible link to pan-coronary syndromes. AB - BACKGROUND: Endothelial apoptosis of atherosclerotic lesions is a possible determinant for the stable-to-vulnerable plaque transition. Recent data support the notion that plaque activation may be a pan-coronary process, advocating the existence of circulating triggers. METHODS AND RESULTS: Serum from 40 healthy subjects (group 1) and 73 patients with stable angina (n=32; group 2) or acute coronary syndromes (n=41; group 3) was incubated with human umbilical vein endothelial cells. The percentage of apoptosis by flow cytometry and Fas, Bax, and Bcl-2 protein expression by immunoblotting were evaluated at entry in patients and control subjects and repeated after 12 months in group 3. At baseline, apoptotic nuclei were higher in group 3 (14+/-6%) than in group 2 (3.3+/-1.8%) and group 1 (1.35+/-0.8%) (P<0.001). Fas and Bcl-2 were increased in group 3 with respect to groups 1 and 2 (P<0.01). Coincubation of group 3 serum with anti-tumor necrosis factor-alpha and anti-interleukin-6 monoclonal antibodies did not affect the human umbilical vein endothelial cell apoptotic process, whereas addition of Trolox decreased apoptosis to <50%. The percentage of apoptosis in group 3 significantly correlated to the numbers of coronary complex lesions at angiography (r=0.58, P<0.0005). In group 3, apoptosis and the Bax/Bcl-2 ratio decreased at 1 year (P<0.0001, P<0.05 respectively). CONCLUSIONS: Serum from patients with acute coronary syndromes displays a proapoptotic effect on human endothelial cells, supporting the theory of the existence of circulating triggers potentially able to activate atherosclerotic lesions. PMID- 12538427 TI - Divergent nitric oxide bioavailability in men and women with sickle cell disease. AB - BACKGROUND: Although reduced endothelial nitric oxide (NO) bioavailability has been demonstrated in arteriosclerotic vascular disease, the integrity of this system in sickle cell disease remains uncertain. METHODS AND RESULTS: We measured forearm blood flow in 21 patients with sickle cell disease (hemoglobin SS genotype) and 18 black control subjects before and after intra-arterial infusions of acetylcholine, nitroprusside, and the NO synthase inhibitor NG-monomethyl-L arginine (L-NMMA). Endothelium-dependent vasodilation, measured by the percent increase in flow induced by acetylcholine infusion, was significantly greater than in controls (252+/-37% for patients versus 134+/-24% for controls; P<0.0001). However, there was a large sex difference in blood flow responses between female and male patients (340+/-46% versus 173+/-41%; P=0.035). Similarly, basal NO bioactivity, as measured by the percent decrease in flow induced by L-NMMA, was depressed in male compared with female patients (-17+/-5% versus -34+/-4%; P=0.01), as was the response to nitroprusside (86+/-21% versus 171+/-22%; P=0.008). L-NMMA reduced the blood flow response to acetylcholine in women, but not in men. Sex differences in vascular cell adhesion molecule-1 were appreciated, with significant correlations between levels of soluble vascular cell adhesion molecule-1 and blood flow responses to L-NMMA and nitroprusside (r=0.53, P=0.004 and r=-0.66, P<0.001, respectively). CONCLUSIONS: NO bioavailability and NO responsiveness are greater in women than in men with sickle cell disease and determines adhesion molecule expression. Endothelium dependent blood flows are largely non-NO mediated in male patients. These results provide a possible mechanism for reported sex differences in sickle cell disease morbidity and mortality and provide a basis for novel pharmacological interventions. PMID- 12538428 TI - Echocardiographic predictors of survival and response to early revascularization in cardiogenic shock. AB - BACKGROUND: Although echocardiography is used in diagnosis and management of myocardial infarction, it has not been established whether specific features of cardiac structure or function early in the course of cardiogenic shock provide prognostic value. The purposes of this substudy of the SHould we emergently revascularize Occluded Coronaries for cardiogenic shocK (SHOCK) trial were to describe the echocardiographic features of cardiogenic shock, identify findings on early echocardiograms associated with outcome, examine the interaction of such features with treatment, and determine whether these features could provide insights into the survival benefit observed with early revascularization and guide selection of patients for this strategy. METHODS AND RESULTS: One hundred seventy-five echocardiograms performed within 24 hours of randomization to the early revascularization (ERV) or initial medical stabilization (IMS) arms of the trial were submitted for quantitative assessment, and 169 were suitable for analysis. The 2 groups were similar in terms of clinical and early echocardiographic characteristics. Mean left ventricular ejection fraction (LVEF) was 31%, and moderate or greater mitral regurgitation (MR) was noted in 39.1%. On multivariate analysis, the only independent predictors of survival were MR severity and LVEF. A survival benefit for the ERV strategy was observed at all levels of LVEF and MR. CONCLUSIONS: A wide range of cardiac structural and functional abnormalities exists in patients presenting with acute cardiogenic shock. Both short- and long-term mortality appear to be associated with initial left ventricular systolic function and MR as assessed by echocardiography, and a benefit of ERV is noted regardless of baseline LVEF or MR. PMID- 12538429 TI - Effect of adrenergic stimulation on action potential duration restitution in humans. AB - BACKGROUND: Enhanced sympathetic activity facilitates complex ventricular arrhythmias and fibrillation. The restitution properties of action potential duration (APD) are important determinants of electrical stability in the myocardium. Steepening of the slope of APD restitution has been shown to promote wave break and ventricular fibrillation. The effect of adrenergic stimulation on APD restitution in humans is unknown. METHODS AND RESULTS: Monophasic action potentials were recorded from the right ventricular septum in 18 patients. Standard APD restitution curves were constructed at 3 basic drive cycle lengths (CLs) of 600, 500, and 400 ms under resting conditions and during infusion of isoprenaline (15 patients) or adrenaline (3 patients). The maximum slope of the restitution curves was measured by piecewise linear regression segments of sequential 40-ms ranges of diastolic intervals in steps of 10 ms. Under control conditions, the maximum slope was steeper at longer basic CLs; eg, mean values for the maximum slope were 1.053+/-0.092 at CL 600 ms and 0.711+/-0.049 at CL 400 ms (+/-SEM). Isoprenaline increased the steepness of the maximum slope of APD restitution, eg, from a maximum slope of 0.923+/-0.058 to a maximum slope of 1.202+/-0.121 at CL 500 ms. The effect of isoprenaline was greater at the shorter basic CLs. A similar overall effect was observed with adrenaline. CONCLUSIONS: The adrenergic agonists isoprenaline and adrenaline increased the steepness of the slope of the APD restitution curve in humans over a wide range of diastolic intervals. These results may relate to the known effects of adrenergic stimulation in facilitating ventricular fibrillation. PMID- 12538430 TI - Vasopressor response to angiotensin II infusion in patients with chronic heart failure receiving beta-blockers. AB - BACKGROUND: A synergistic interaction between the angiotensin II (Ang II) type 1 receptor and alpha1-adrenergic receptors has been described. We hypothesized that the nonselective beta-antagonist carvedilol, through its alpha1-adrenergic blocking properties, may modulate vascular reactivity to Ang II in patients with chronic heart failure (CHF). Accordingly, we compared the vasopressor response to infused Ang II in patients treated with carvedilol and metoprolol, a selective beta-antagonist. METHODS AND RESULTS: All subjects were treated with carvedilol or metoprolol for at least 3 months. ACE inhibitor therapy was standardized to enalapril 40 mg/d or the maximally tolerated dose. Exogenous Ang II was administered as sequential intravenous bolus injections (2.5 to 30 ng/kg) titrated to a rise in radial artery systolic pressure of > or =20 mm Hg. The dose of Ang II required to elicit a change of 20 mm Hg in radial artery systolic pressure (PD20) defined the vasopressor response to Ang II. Twenty subjects with CHF (mean left ventricular ejection fraction 28+/-9%, New York Heart Association class II [n=13] and III [n=7]) were studied. There was no correlation between plasma Ang II levels and PD20. However, the PD20 was significantly higher in patients treated with carvedilol than in those treated with metoprolol (20 [range 2.5 to 30] versus 5 [range 2.5 to 10] ng/kg, P=0.019). CONCLUSIONS: The vasopressor response to Ang II infusion in patients treated with carvedilol was significantly lower than in patients treated with metoprolol. Whether this is due to the alpha1-adrenergic blocking or other ancillary properties of carvedilol warrants further investigation. PMID- 12538431 TI - Effect of erythropoietin on exercise capacity in patients with moderate to severe chronic heart failure. AB - BACKGROUND: Patients with chronic heart failure (CHF) are frequently anemic. An increase in hemoglobin could enhance exercise performance by increasing oxygen delivery. We investigated the effect of erythropoietin (EPO) on exercise performance in anemic patients with CHF. METHODS AND RESULTS: Twenty-six anemic patients aged 57+/-11 years were randomized to receive EPO (15 000 to 30 000 IU per week) or placebo for 3 months. Parameters measured at baseline and end therapy included blood parameters (hemoglobin, hematocrit, plasma volume), exercise parameters (peak oxygen consumption [VO2], exercise duration, 6-minute walk), muscle aerobic metabolism (half-time of VO2 and near infrared recovery), and forearm vasodilatory function. EPO was well tolerated by all patients. Twelve patients in the EPO group felt improvement versus 1 in the placebo group (P<0.05). There were significant increases in hemoglobin (11.0+/-0.5 to 14.3+/ 1.0 g/dL, P<0.05), peak VO2 (11.0+/-1.8 to 12.7+/-2.8 mL. min(-1) x kg(-1), P<0.05) and exercise duration (590+/-107 to 657+/-119 s, P<0.004) in the EPO group but no significant changes in the control group. Resting and hyperemic forearm vascular resistance and indices of the rate of muscle oxidative capacity were unchanged in both groups. CONCLUSION: EPO significantly enhances exercise capacity in patients with CHF. One mechanism of improvement in VO2 is increased oxygen delivery from increased hemoglobin concentration. PMID- 12538433 TI - Magnetocardiographic rhythm patterns at initiation and termination of fetal supraventricular tachycardia. AB - BACKGROUND: Using fetal magnetocardiography (fMCG), we characterize for the first time the electrophysiological patterns of initiation and termination of reentrant fetal supraventricular tachycardia (SVT), the most common form of life threatening fetal arrhythmia. METHODS AND RESULTS: In contrast to the expectation that reentrant SVT is initiated by spontaneous premature atrial contractions (PACs) and is terminated by spontaneous block, 5 distinct patterns of initiation and 4 patterns of termination were documented, with the most common patterns of initiation involving reentrant PACs. Waveform morphology and timing, including QRS and ventriculoatrial interval, were assessed. This enabled detection of such phenomena as Wolff-Parkinson-White syndrome, QRS aberrancy, and multiple reentrant pathways that were crucial for defining the rhythm patterns. In addition, fMCG actocardiography revealed an unexpectedly strong association between fetal trunk movement and the initiation and termination of SVT, suggesting that autonomic influences play a key role. CONCLUSIONS: This study demonstrates that the patterns of initiation and termination of fetal SVT are more diverse than is generally believed and that the most common patterns of initiation involve reentrant PACs. The ability to discern such patterns can help elucidate the underlying mechanisms and guide antiarrhythmic drug therapy. fMCG provides a noninvasive means of analyzing complex tachyarrhythmia in utero, with efficacy approaching that of postnatal electrocardiographic rhythm monitoring. PMID- 12538432 TI - Skeletal muscle reflex in heart failure patients: role of hydrogen. AB - BACKGROUND: An important role of the increased stimulation of skeletal muscle ergoreceptors (intramuscular afferents sensitive to products of muscle work) in the genesis of symptoms of exertion intolerance in chronic heart failure (CHF) has been proposed. With the use of selective infusions and dietary manipulation methods, we sought to identify the role of H+, K+, lactate, and peripheral hemodynamics on ergoreflex overactivation. METHODS AND RESULTS: Ten stable CHF patients (aged 67.9+/-2.5 years, peak oxygen uptake 16.3+/-1.2 mL x kg(-1) x min( 1)) and 10 age-matched and sex-matched healthy subjects were studied. The ergoreflex contribution to ventilation was assessed by post-handgrip regional circulatory occlusion (PH-RCO) and computed as the difference in ventilation between PH-RCO and a control run without PH-RCO. This test was performed on 6 separate occasions. On each occasion a different chemical was infused (insulin, sodium nitroprusside, sodium bicarbonate, dopamine, or saline) or a 36-hour glucose-free diet was undertaken before the test. During all stages of the protocol, the local muscular blood effluent concentrations of H+, K+, glucose, and lactate were assessed. An ergoreflex effect on the ventilatory response was seen in patients (versus control subjects) during the saline infusions (6.7+/-2.3 L/min versus -0.1+/-0.5 L/min, P<0.01). The only intervention to significantly lower the ergoreflex was sodium bicarbonate (0.4+/-0.3 L/min versus -0.2+/-0.4 L/min in control subjects, P=NS; versus saline P<0.05), which also reduced H(+) concentration during exercise (47.4+/-1.3 versus 50.0+/-1.4 nmol/L on saline, P<0.05). CONCLUSION: A reduction of the H+ concentration by infusion of sodium bicarbonate abolishes the increased ergoreceptor activity in CHF, suggesting a role of H+ in ergoreflex activation, either directly or indirectly. PMID- 12538434 TI - Inhibition of experimental intimal thickening in mice lacking a novel G-protein coupled receptor. AB - BACKGROUND: Vascular restenosis attributable to intimal thickening remains a major problem after percutaneous transluminal coronary angioplasty (PTCA). METHODS AND RESULTS: Through differential-display analysis, we have identified a novel gene whose expression was increased after catheter injury of rabbit aorta. The gene that is expressed predominantly in vascular smooth muscle cells encodes a novel protein with 7 transmembrane domains, and we termed it ITR (intimal thickness-related receptor). The ITR sequence contains a motif common to the Rhodopsin-like GPCR (G-protein-coupled receptor) superfamily. In vivo analyses of this gene revealed that expression of ITR protein increased with intimal thickening induced by cuff placement around murine femoral artery. Furthermore, ITR-knockout mice were found to be resistant to this experimental intimal thickening. CONCLUSIONS: ITR thus seems to be a novel receptor that may play a role in vascular remodeling and that may represent a good target for development of drugs in the prevention of vascular restenosis. PMID- 12538435 TI - Interleukin-6-deficient mice resist development of autoimmune myocarditis associated with impaired upregulation of complement C3. AB - BACKGROUND: Interleukin (IL)-6 regulates various aspects of the immune response. In the context of heart diseases, it has been recognized as a prognostic factor for dilated cardiomyopathy, which often results from myocarditis. METHODS AND RESULTS: Using IL-6-deficient mice, we studied the role of IL-6 in a model of autoimmune myocarditis resulting from immunization with a peptide derived from cardiac alpha-myosin. Prevalence and severity of myocarditis were markedly reduced in the absence of IL-6. CD4+ T cells from immunized IL-6-deficient mice proliferated poorly on restimulation with specific antigen in vitro and did not mediate disease on adoptive transfer into IL-6-competent RAG-2-deficient mice, which otherwise lack B cells and T cells. Production of complement C3, a crucial factor for the development of myocarditis, was strongly upregulated in IL-6+/+ but not in IL-6-deficient mice after immunization. CONCLUSIONS: Our results demonstrate that IL-6 is required for the expansion of autoimmune CD4+ T cells and the pathogenesis of autoimmune myocarditis, possibly by upregulation of complement C3. PMID- 12538437 TI - Increased medial degradation with pseudo-aneurysm formation in apolipoprotein E knockout mice deficient in tissue inhibitor of metalloproteinases-1. AB - BACKGROUND: The tissue inhibitor of metalloproteinases-1 (TIMP-1) is expressed in atherosclerotic lesions, where it may play a critical role in regulating the activity of matrix metalloproteinases (MMPs). Several MMPs are overexpressed in the atherosclerotic plaque, and they are believed to contribute to the expansion and rupture of the lesion. METHODS AND RESULTS: The Timp-1-knockout mouse model (Timp-1-/-) was crossed into the apolipoprotein E-knockout (apoE0) background. A study population of male apoE0 mice, half of them deficient in TIMP-1, was fed an atherogenic diet. After 10 weeks of the diet, the mean lesion sizes of the two groups of animals were not significantly different, and the average content of fibrillar collagen and macrophages in the lesions was similar. There was no sign of plaque hemorrhage, even after 22 weeks of high-fat diet, indicating that deficiency in TIMP-1 does not predispose to luminal rupture. However the atherosclerotic lesions of the Timp-1-/0 mice developed more aortic medial ruptures, in which all elastic lamellae of the media were degraded and infiltrated with macrophages, forming pseudo-microaneurysms. After 10 weeks of high-fat diet, the Timp-1-/0/apoE0 mice averaged 1.9+/-1.2 medial ruptures in the proximal aorta, compared with 0.5+/-0.7 for the apoE0 controls (P<0.003). At the site of degradation, in situ zymography revealed that the gelatinolytic activity, mainly associated with macrophages, could be abolished by the addition of MMP inhibitors. CONCLUSIONS: These data strongly suggest that TIMP-1 plays a key role in preventing medial degradation associated with atherosclerosis through its ability to inhibit the MMPs that are involved in the disruption of the media. PMID- 12538436 TI - Positron-emission tomography reporter gene expression imaging in rat myocardium. AB - BACKGROUND: This study examines the quantitative accuracy, detection sensitivity, and time course of imaging the expression of a mutant herpes simplex type-1 virus thymidine kinase (HSV1-sr39tk) PET reporter gene in rat myocardium by using the PET reporter probe 9-(4-[18F]-Fluoro-3-Hydroxymethylbutyl)-Guanine ([18F]-FHBG) and a small-animal PET (microPET). METHODS AND RESULTS: In 40 rats, adenovirus expressing HSV1-sr39tk driven by a cytomegalovirus promoter (Ad-CMV-HSV1-sr39tk, 1x10(6) to 1x10(9) pfu) was injected through a thoracotomy directly into the left ventricular myocardium. After 3 days, myocardial perfusion was imaged with [13N] ammonia for delineating the left ventricular myocardium, followed by imaging the expression of the reporter gene with intravenous [18F]-FHBG. The total myocardial [18F]-FHBG accumulation was quantified in percent of injected dose (%ID). Immunohistochemistry and autoradiography demonstrated HSV1-sr39tk enzyme (HSV1 sr39TK) and accumulation of [18F]-FHBG in the inoculated myocardium in 3 rats each. In 24 rats with various viral titers, the %ID was correlated with ex vivo well counting (r2=0.981, P<0.0001) and myocardial HSV1-sr39TK activity by tissue enzyme activity assay (r2=0.790, P<0.0001). Myocardial [18F]-FHBG accumulation was identified at viral titers down to 1x10(7) pfu. In 6 rats serially imaged up to day 17, myocardial [18F]-FHBG accumulation on microPET peaked on days 3 to 5 and was no longer identified on days 10 to 17. CONCLUSIONS: HSV1-sr39tk reporter gene expression can be monitored with [18F]-FHBG and microPET in rat myocardium quantitatively and serially with high detection sensitivity. Cardiac PET reporter gene imaging offers the potential of monitoring the expression of therapeutic genes in cardiac gene therapy. PMID- 12538438 TI - Altered temporal profile of heat shock factor 1 phosphorylation and heat shock protein 70 expression induced by heat shock in nucleus tractus solitarii of spontaneously hypertensive rats. AB - BACKGROUND: We demonstrated recently that heat shock (HS)-induced heat shock protein 70 (HSP70) expression in bilateral nucleus tractus solitarii (NTS), the terminal site in the brain stem for primary baroreceptor afferents, confers cardiovascular protection against heatstroke by potentiating baroreceptor reflex (BRR) response. This study evaluated the hypothesis that altered regulation of HSP70 expression may be associated with the heightened susceptibility to heatstroke during hypertension. METHODS AND RESULTS: Spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats anesthetized with propofol were used. Compared with WKY rats, significant induction in HSP70 or phosphorylation of heat shock factor 1 (HSF1), but not HSF2, in the NTS and potentiation of BRR response in SHR occurred earlier (4 versus 8 hours), reaching peak magnitude sooner (16 versus 24 hours), and declined more rapidly after a brief hyperthermic HS (42+/-0.5 degrees C for 15 minutes). The protection conferred by HS against hypotension and bradycardia during the onset of heatstroke (45 degrees C for 60 minutes), although effective, was less effective in SHR. Microinjection bilaterally into the NTS of the selective protein kinase A (PKA) inhibitor H-89 (100 pmol) or the selective PKC inhibitor calphostin C (100 pmol) significantly attenuated all of the above events induced in SHR by HS. However, only H-89 was effective in WKY rats. CONCLUSIONS: An altered temporal profile of HS-induced HSP70 expression or potentiation of BRR response by concurrent activation via both PKA and PKC pathways of phosphorylation of HSF1 in the NTS may be associated with greater susceptibility to heatstroke during hypertension. PMID- 12538439 TI - Arterial and cardiac aging: major shareholders in cardiovascular disease enterprises: Part II: the aging heart in health: links to heart disease. PMID- 12538440 TI - Images in cardiovascular medicine. Tumor-like calcification of the mitral annulus: diagnosis with multislice computed tomography. PMID- 12538441 TI - Cardiology patient pages. Cardiovascular disease in the diabetic patient. PMID- 12538442 TI - Images in cardiovascular medicine. Optimal guidance for intimal flap fenestration in aortic dissection by transvenous two-dimensional and Doppler ultrasonography. PMID- 12538443 TI - Food and Drug Administration: Cardiovascular and Renal Drugs Advisory Committee, 98th meeting, January 6th-7th, 2003. PMID- 12538444 TI - Erythropoietin--not at the Olympics but maybe for anemic heart failure patients. PMID- 12538445 TI - Peroxisome proliferator-activated receptor gamma and cancers. AB - The peroxisome proliferator-activated receptor gamma (PPARgamma) ligands have anticancer activity against a wide variety of neoplastic cells in vitro. Animal studies have chronicled their in vivo anticancer effects and chemopreventive capabilities. In addition, moderate anticancer activities of PPARgamma ligands with minimal toxicities have been observed in patients with liposarcomas and prostate cancers. These compounds can slow growth and induce partial differentiation of selected cancer cells. They can decrease levels of cyclin D(1) and E, inflammatory cytokines, and nuclear factor kappaB and increase expression of p21(waf1) and p27(kip1). Surprisingly, some or many of these effects may occur independently of PPARgamma. Other data suggest that PPARgamma may behave as a tumor suppressor gene, although several compelling murine models, paradoxically, suggest that under selected circumstances, PPARgamma ligands may stimulate cancer formation. Nevertheless, the bulk of studies showed that PPARgamma ligands do have antiproliferative activity against many transformed cells and may be helpful in the setting of adjuvant and chemopreventive treatments of several common tumors, including colon, prostate, and breast cancers. PMID- 12538446 TI - The statins as anticancer agents. AB - 3-Hydroxy-3-methylgutaryl CoA reductase inhibitors, commonly referred to as the statins, have proven therapeutic and preventative effects in cardiovascular diseases. Recently, there are emerging interests in their use as anticancer agents based on preclinical evidence of their antiproliferative, proapoptotic, anti-invasive, and radiosensitizing properties. Inhibition of 3-hydroxy-3 methylgutaryl CoA reductase by the statins interferes with the rate-limiting step of the mevalonate pathway, leading to reduced levels of mevalonate and its downstream products, many of which play important roles in critical cellular functions such as membrane integrity, cell signaling, protein synthesis, and cell cycle progression. Perturbations of these processes in neoplastic cells by the statins may therefore result in control of tumor initiation, growth, and metastasis. The statins have demonstrated growth inhibitory activity in cancer cell lines and preclinical tumor models in animals. Phase I trials of statins in humans have demonstrated myotoxicity as their main dose-limiting toxicity, and Phase II trials in various tumor types are ongoing to evaluate their efficacy. Potential future directions in the development of the statins as anticancer agents include combinations with chemotherapeutic or other molecular-targeted agents, combinations with radiotherapy, maintenance therapy in minimal disease status, and as chemopreventive therapy. PMID- 12538447 TI - Prevention of ovarian cancer: intraepithelial neoplasia. AB - To reduce the incidence and mortality associated with invasive cancers, the Intraepithelial Neoplasia (IEN) Task Force recommends that carcinogenesis be viewed as a disease that requires treatment. This publication outlines the current knowledge of IEN of the ovary and reviews chemoprevention possibilities for ovarian cancer. Ovarian cancer has the highest mortality of all of the gynecological cancers and is the fourth leading cause of death from cancer in women. The IEN Task Force has defined precancer as a noninvasive lesion that has genetic abnormalities, loss of cellular control functions, and some phenotypic characteristics of invasive cancer with a substantial likelihood of developing invasive cancer. The IEN Task Force recommends targeting moderate to severe dysplasia for new IEN treatment agents in clinical trials. Ovarian cancer does not have a clear preinvasive lesion yet merits considerable study for new prevention strategies because of the high mortality associated with ovarian cancer. There is a great unmet clinical need for treatments that can prevent ovarian cancer by providing nonsurgical options that treat the entire epithelial layer. New prevention strategies hold significant promise to reduce the mortality from ovarian cancer. PMID- 12538448 TI - Commentary re: Q-T Le et al, identification of osteopontin as a prognostic marker for head and neck squamous cell carcinomas. clin cancer res, 9: 31-32, 2003. PMID- 12538449 TI - Persistent replication of the modified chimeric adenovirus ONYX-015 in both tumor and stromal cells from a patient with gall bladder carcinoma implants. AB - PURPOSE: ONYX-015 is a chimeric, E1B-deleted adenovirus designed to replicate preferentially in p53-deficient tumor cells; however, little is understood about its actual replication potential in human tumors. We hypothesized that replication of a late viral gene, hexon, would demonstrate replication of virus in human tissues. EXPERIMENTAL DESIGN: In the course of a clinical trial, a patient with paired abdominal wall implants from a primary gall bladder carcinoma was injected with ONYX-015, 1 x 10(10) viral particles/lesion, followed by sequential excision of the lesions at 37 h and 7 days. Tissue sections were analyzed for evidence of viral replication. RESULTS: In situ Reverse transcription-PCR was used to measure expression of hexon. Strong signals were obtained in gland-forming tumor cells both at 37 h and at 7 days. Signal was predominantly observed in the cytoplasm. The signal was also observed in adjacent normal stromal cells. Analysis of p53 status of the tumor by immunohistochemistry and Affymetrix Genechip demonstrated an inactivating mutation in p53. Routine H&E staining of the tumor sections revealed no evidence of necrosis at 37 h or 7 days after injection of virus. Presence of viral protein at both 37 h and 7 days was confirmed by immunohistochemistry using antibodies directed against hexon, penton, and fiber proteins. CONCLUSIONS: Evidence for replication of hexon confirms that ONYX-015 is not only present but capable of replicating in tumor cells up to 1 week after intralesional injection and that replication is not confined to p53-mutated tumor cells. PMID- 12538450 TI - The granulin-epithelin precursor/PC-cell-derived growth factor is a growth factor for epithelial ovarian cancer. AB - PURPOSE: The role of growth factors in ovarian cancer development and progression is complex and multifactorial. We hypothesized that new growth factors may be identified through the molecular analysis of ovarian tumors as they exist in their native environment. EXPERIMENTAL DESIGN: RNA extracted from microdissected serous low malignant potential (LMP) and invasive ovarian tumors was used to construct cDNA libraries. A total of 7300 transcripts were randomly chosen for sequencing, and those transcripts were statistically evaluated. Reverse transcription-PCR and immunohistochemistry were used to validate the findings in tumor tissue samples. Ovarian cancer cell lines were used to test gene effects on monolayer growth, proliferative capacity, and density-independent growth. RESULTS: Analysis of the pooled library transcripts revealed 26 genes differentially expressed between LMP and invasive ovarian cancers. The granulin epithelin precursor [GEP/PC-cell derived growth factor (PCDGF)] was expressed only in the invasive ovarian cancer libraries (P < 0.028) and was absent in the LMP libraries (0 of 2872 clones). All of the invasive tumor epithelia, 20% of the LMP tumor epithelia, and all of the stroma from both subsets expressed GEP by reverse transcription-PCR. Immunohistochemical staining for GEP was diffuse and cytosolic in invasive ovarian cancer tumor cells compared with occasional, punctate, and apical staining in LMP tumor epithelia. Antisense transfection of GEP into ovarian cancer cell lines resulted in down-regulation of GEP production, reduction in cell growth (P < 0.002), decrease in the S-phase fraction (P < 0.04), and loss of density-independent growth potential (P < 0.01). CONCLUSION: cDNA library preparation from microdissected tumor epithelium provided a selective advantage for the identification of growth factors for epithelial ovarian cancer. Differential granulin expression in tumor samples and the antiproliferative effects of its antisense down-regulation suggest that GEP may be a new autocrine growth factor and molecular target for epithelial ovarian cancer. PMID- 12538451 TI - Deletions in chromosome arms 3p and 11q are new prognostic markers in localized and 4s neuroblastoma. AB - PURPOSE: To find new nonrandom chromosomal changes in neuroblastoma (NB) with a potential to forecast the patient's outcome, alterations in chromosome arms 3p and 11q were investigated. EXPERIMENTAL DESIGN: Frequency and prognostic potential of 3p and 11q alterations in 144 NBs were analyzed using interphase fluorescence in situ hybridization with DNA probes for 3p26 and 11q23. Aberrations were defined as deletion (monosomy of a specific region) or imbalance (at least two intact and additional 3p26- or 11q23-deleted chromosomes). RESULTS: Forty-two of 144 cases (29%) displayed 11q alterations (21% deletions, 8% imbalances). Most aberrations were associated with stage 4 disease (28 of 59, 47%) but were also present in localized and 4s tumors (14 of 85, 16%; P = 0.007). Patients with 11q deletion/imbalance were significantly older at diagnosis (P < 0.001). Changes in 3p were detected in 26 of 144 (18%) samples (15% deletions, 3% imbalances). These alterations were also associated with stage 4 [20 of 59 (34%) versus 6 of 85 (7%) in stages 1-3 and 4s, P = 0.007], and the median age was increased (P < 0.001). Aberrations in both chromosomes were highly associated with each other (P < 0.001). MYCN amplification (MNA) was detected in 10% and 12% of tumors with 11q and 3p alterations, and changes in 1p36 occurred in 13% and 26% of the 3p- and 11q-aberrant tumors. MYCN amplification and 11q deletion/imbalance tended to show an inverse correlation (P = 0.07) as well as 1p and 3p deletion/imbalance (P = 0.07). Patients with 3p and 11q abnormalities in localized/4s tumors showed an inferior outcome compared with those without these alterations (P = 0.002 and P = 0.0027, respectively), in particular in MYCN single copy tumors (P < 0.0001 and P = 0.0006, respectively). CONCLUSION: Alterations in 3p and 11q are frequent nonrandom aberrations in NB and define a new high-risk subgroup in MYCN single copy stage 1-3 and 4s disease. PMID- 12538452 TI - Identification of osteopontin as a prognostic plasma marker for head and neck squamous cell carcinomas. AB - PURPOSE: Tumor hypoxia modifies treatment efficacy and promotes tumor progression. Here, we investigated the relationship between osteopontin (OPN), tumor pO(2), and prognosis in patients with head and neck squamous cell carcinomas (HNSCC). EXPERIMENTAL DESIGN: We performed linear discriminant analysis, a machine learning algorithm, on the NCI-60 cancer cell line microarray expression database to identify a gene profile that best distinguish cell lines with high Von-Hippel Lindau (VHL) gene expression, an important regulator of hypoxia-related genes, from those with low expression. Plasma OPN levels in 15 volunteers, 31 VHL patients, and 54 HNSCC patients were quantitatively measured by ELISA. The relationships between plasma OPN levels, tumor pO(2) as measured by the Eppendorf microelectrode, freedom from relapse (FFR), and survival in HNSCC patients were evaluated. RESULTS: Microarray analysis indicated that OPN gene expression inversely correlated with that of VHL. These findings were confirmed by Northern blot analysis. ELISA studies and Western blot in a HNSCC cell line demonstrated that hypoxia exposure resulted in increased OPN secretion. Patients with VHL syndrome had significantly higher plasma OPN levels than healthy volunteers. Plasma OPN level inversely correlated with tumor pO(2) (P = 0.003, r = -0.42). OPN levels correlated with clinical outcomes. The 1-year FFR and survival rates were 80 and 100%, respectively, for patients with OPN levels 450 ng/ml (P = 0.002 and 0.0005). Multivariate analysis revealed that OPN was an independent predictor for FFR and survival. CONCLUSIONS: Plasma OPN levels appeared to correlate with tumor hypoxia in HNSCC patients and may serve as noninvasive tests to identify patients at high risk for tumor recurrence. PMID- 12538453 TI - Hepatocyte growth factor receptor, matrix metalloproteinase-11, tissue inhibitor of metalloproteinase-1, and fibronectin are up-regulated in papillary thyroid carcinoma: a cDNA and tissue microarray study. AB - PURPOSE: To screen and validate the global gene expression in papillary thyroid carcinoma (PTC) using cDNA expression arrays and immunohistochemistry on tumor tissue microarrays in an attempt to find genes that may be of importance in the molecular pathogenesis and malignant progression of PTC. EXPERIMENTAL DESIGN: Eighteen PTC tissue specimens were compared with three morphologically normal thyroid specimens by applying Atlas Human Cancer 1.2 Array membranes printed with cDNAs of 1176 human genes involved in cancer. Results for selected genes were confirmed by reverse transcription-PCR. Protein expression of selected genes was further studied using a tissue microarray consisting of 107 PTCs and compared with histologically normal thyroid tissue samples. RESULTS: By cDNA arrays, two genes [c-MET and matrix metalloproteinase (MMP)-11] were expressed only in tumor tissue, where they were present in >50% of cases. Ten genes [macrophage inhibitory cytokine-1, CGD, fibronectin (FN), hypoxia-inducible factor 1, Fc epsilon-receptor gamma-chain, lactate dehydrogenase A, HLA-DBP1, AH receptor, tissue inhibitor of metalloproteinase (TIMP-1), and glycyl-tRNA-synthetase] were found to be up-regulated >2-fold in 40-100% of cancers, whereas 9 genes (GADD153, polykystic kidney disease-1, CYR61, DPC4, HBA1, gravin, DLG3, protein tyrosine phosphatase sigma, and heterochromatin protein 1 homologue-alpha) were down regulated to <50% of their normal levels in 40-94% of cases. Conventional reverse transcription-PCR gave consistent results with the cDNA array findings for all four genes selected to be studied (c-MET, FN, TIMP-1, and GADD153). Immunohistochemistry for three selected proteins, FN, MMP-11, and TIMP-1, showed positive staining in 81, 87, and 68% of the tumor samples, respectively. CONCLUSIONS: Several novel and previously undetected tumor promoting/inhibiting genes may be of importance in the molecular pathogenesis and malignant progression of PTC. Transcription of these genes may result in overexpression of proteins, such as c-MET, MMP-11, TIMP-1, and FN, which may contribute to the pathogenesis of PTC. PMID- 12538454 TI - Repeated administrations of interleukin (IL)-12 are associated with persistently elevated plasma levels of IL-10 and declining IFN-gamma, tumor necrosis factor alpha, IL-6, and IL-8 responses. AB - PURPOSE: Repeated administrations of recombinant human interleukin-12 (rHuIL-12) to cancer patients are characterized by a reduction of side effects during treatment. Induction of IFN-gamma, considered a key mediator of antitumor effects of IL-12, is known to decline on repeated administrations. We studied whether other immunological effects of rHuIL-12 are tapered in the course of treatment. EXPERIMENTAL DESIGN: In a Phase I study of 26 patients with advanced renal cell cancer, rHuIL-12 was administered s.c. on day 1, followed by 7 days rest and six injections administered over a 2-week time period. Plasma concentrations of various cytokines were monitored, as well as absolute counts of circulating leukocyte and lymphocyte subsets. RESULTS: The first injection of IL-12 was accompanied by rapid, transient, and dose-dependent increments of plasma levels IFN-gamma, tumor necrosis factor-alpha, IL-10, IL-6, IL-8, but not IL-4, as well as rapid, transient, and dose-dependent reductions of lymphocyte, monocyte, and neutrophil counts. The major lymphocyte subsets, i.e., CD4+ and CD8+ T cells, B cells, and natural killer cells, followed this pattern. On repeated rHuIL-12 injections, IL-10 concentrations increased further, whereas the transient increments of IFN-gamma, tumor necrosis factor-alpha, IL-6, and IL-8 concentrations, as well as the fluctuations of the leukocyte subset counts, were tapered. Dose escalation of IL-12 within clinically tolerable margins did not reduce the decline of these immunological effects. CONCLUSIONS: Induction of pro inflammatory cytokines and associated fluctuations in leukocyte subset counts decrease on repeated administrations of rHuIL-12. The steady increment of IL-10 plasma levels may mediate the observed down-regulation of clinical and immunological effects. PMID- 12538455 TI - Low-dose IFN-gamma induces tumor MHC expression in metastatic malignant melanoma. AB - Specific antitumor immune responses require expression of MHC class I or II molecules on tumor cells, and MHC antigen down-regulation is a presumed tumor growth promoting mechanism. Because IFN-gamma up-regulates tumor MHC antigen expression in vitro, in this Phase II trial of an immunologically active dose and schedule we evaluated whether this was the case in vivo. Twenty-three patients with metastatic melanoma were treated with IFN-gamma 100 microg/m(2) s.c. once weekly for a maximum of 6 months. There were three complete responses, now maintained for 53, 36, and 25 months. The remainder had progressive disease. The treatment was well tolerated, with no toxicity exceeding National Cancer Institute Common Toxicity Criteria grade II. Immunohistochemical analysis of tumor biopsies during treatment was performed using monoclonal antibodies to HLA class I (W/632) and class II (CR3/43) monomorphic determinants. HLA class I was down-regulated in 2 of 19 patients pretreatment and up-regulated by IFN-gamma in both. HLA class II was down-regulated pretreatment in 14 of 18 patients and up regulated by IFN-gamma in 6 (43%). The HLA up-regulation persisted throughout the study. IFN-gamma induced significant but short-lived up-regulation of surrogate markers of monocyte activation (serum neopterin) and class I up-regulation (serum beta-2-microglobulin) in most patients. There was no consistent relationship between surrogate marker up-regulation, tumor antigen up-regulation, and responses. The study shows that the significant immune modulation induced by IFN gamma does not correlate with tumor responses and that the serum surrogate marker changes do not reflect tumor events. The durable and long-lived responses, clear demonstration of tumor MHC up-regulation, and low toxicity suggest that weekly IFN-gamma 100 microg/m(2) would be a useful addition to chemoimmunotherapeutic regimens. PMID- 12538457 TI - Phase I and pharmacokinetic study of escalating dose of docetaxel administered with granulocyte colony-stimulating factor support in adult advanced solid tumors. AB - PURPOSE: The purpose of our study was to assess the feasibility, toxicity, and pharmacokinetics of an escalating dose of docetaxel when administered with granulocyte colony-stimulating factor (G-CSF) support every 3 weeks. EXPERIMENTAL DESIGN: Patients with advanced solid malignancies were treated with escalating doses of docetaxel as a 1-h infusion every 3 weeks, supported by s.c. administration of human recombinant glycosilated G-CSF Granocyte (lenograstim), 5 microg/kg/day (from day 4 until neutrophil count >0.5-10(g)/liter for two consecutive days). Plasma sampling was performed to characterize the pharmacokinetics of docetaxel at the new recommended high-dose level. RESULTS: Forty-seven patients were treated with 116 courses of docetaxel at eight dose levels ranging from 100-185 mg/m(2). Dose-limiting toxicities were nonhematologic and included mucositis and dermatitis. Severe skin toxicity observed at 185 mg/m(2) led to discontinuing the study, and 175 mg/m(2) was selected as the recommended dose of docetaxel + G-CSF for future Phase II studies. Analysis of multiple courses revealed dermatitis, mucositis, arthralgia/myalgia, and neuropathy as the main dose-related toxic events. At 175 mg/m(2) mean +/- SD values for docetaxel plasmatic peak, area under the curve, clearance, volume of distribution, and terminal half-life were 6.7 +/- 1.7 microg/ml, 9.7 +/- 4 microg.h/ml, 34.2 +/- 12 liters/h, and 122.7 +/- 124 liters, respectively. Of the 16 patients treated at 175 mg/m(2), 8 patients responded (7 breast cancer and 1 lung cancer patients) including one complete response (1 breast cancer patient). CONCLUSIONS: Using G-CSF support allows substantial dose escalation of docetaxel. Whether such a dose increase improves the response rate warrants further investigation. At the highest dose level studied, pharmacokinetic parameters seem to maintain a linear profile. PMID- 12538456 TI - Induction of p53-regulated genes and tumor regression in lung cancer patients after intratumoral delivery of adenoviral p53 (INGN 201) and radiation therapy. AB - PURPOSE: We designed a prospective single arm Phase II study to evaluate the feasibility and mechanisms of apoptosis induction after Ad-p53 (INGN 201) gene transfer and radiation therapy in patients with non-small cell lung cancer. EXPERIMENTAL DESIGN: Nineteen patients with nonmetastatic non-small cell lung cancer who were not eligible for chemoradiation or surgery were treated as outpatients with radiation therapy to 60 Gy over 6 weeks in conjunction with three intratumoral injections of Ad-p53 (INGN 201) on days 1, 18, and 32. RESULTS: Seventeen of 19 patients completed all planned radiation and Ad-p53 (INGN 201) gene therapy as outpatients. The most common adverse events were grade 1 or 2 fevers (79%) and chills (53%). Three months after completion of therapy, pathologic biopsies of the primary tumor revealed no viable tumor (12 of 19 patients, 63%), viable tumor (3 of 19 patients, 16%), and not assessed (4 of 19 patients, 21%). Computed tomography and bronchoscopic findings at the primary injected tumor revealed complete response (1 of 19 patients, 5%), partial response (11 of 19 patients, 58%), stable disease (3 of 19 patients, 16%), progressive disease (2 of 19 patients, 11%), and not evaluable (2 of 19 patients, 11%). Quantitative reverse transcription-PCR analysis of the four p53 related genes [p21 (CDKN1A), FAS, BAK, and MDM2] revealed that Bak expression was increased significantly 24 h after Ad-p53 (INGN 201) injection and levels of CDKN1A and MDM2 expression were increased over the course of treatment. CONCLUSIONS: Intratumoral injection of Ad-p53 (INGN 201) in combination with radiation therapy is well tolerated and demonstrates evidence of tumor regression at the primary injected tumor. Serial biopsies of the tumor suggest that BAK gene expression is most closely related to Ad-p53 (INGN 201) gene transfer. PMID- 12538458 TI - Weekly bryostatin-1 in metastatic renal cell carcinoma: a phase II study. AB - PURPOSE: We conducted a Phase II trial of bryostatin-1, an inhibitor of protein kinase C, in advanced renal cell carcinoma to measure toxicity, response rate, time to progression, and induction of cytokines. EXPERIMENTAL DESIGN: A total of 32 patients (26 male and 6 female) received bryostatin-1 at 35-40 microg/m(2) i.v. over 1 h on days 1, 8, and 15 of each 4-week cycle. Plasma interleukin-6, tumor necrosis factor-alpha, and C-reactive protein levels were assayed pretreatment, 1 and 23 h after completion of bryostatin-1 infusion at weeks 1 and 5. RESULTS: Cycles (102) of bryostatin-1 were given (median 2, range 1-8). The most common grade 1 or 2 toxicities were myalgias (46.8%), fatigue (59.3%), and dyspnea (18.8%). Grade 3-4 toxicity included myalgias (40.6%), ataxia (9.3%), and dyspnea (15.6%). Four (12%) patients experienced cardiac events while on study (cardiac arrhythmias and congestive heart failure occurred in 2 patients, and 2 patients had fatal cardiac arrests). Of 32 patients evaluable for response, 2 (6.3%) had partial responses lasting 9 with 6 months. A total of 15 patients (46.8%) had stable disease, and 6 (18.8%) patients had stable disease for >or=6 months. Plasma interleukin-6 increased >or=2-fold over baseline measurements in 5 of 17 patients (29.4%) but did not correlate with response or toxicity. CONCLUSIONS: Although weekly bryostatin-1 at 35-40 microg/m(2) produced a low proportion of objective responses, prolonged (>6 months) stable disease or partial remission in 25% of patients suggests that this agent, or other inhibitors of protein kinase C, may have a role in the treatment of renal cell carcinoma, perhaps in combination with other agents. PMID- 12538459 TI - A phase I trial of ISIS 2503, an antisense inhibitor of H-ras, in combination with gemcitabine in patients with advanced cancer. AB - PURPOSE: The purpose of this study was to define the toxicity, pharmacokinetics, and clinical activity of the combination of ISIS 2503, an oligodeoxynucleotide antisense inhibitor of H-ras, and gemcitabine in patients with advanced solid tumors. EXPERIMENTAL DESIGN: The target dose of ISIS 2503 on this study was 6 mg/kg/day. Twenty-seven patients (16 male, 11 female) received 97 treatment courses (median, 2; range, 1-13). Nineteen patients were treated with a fixed gemcitabine dose of 1000 mg/m(2) on days 1 and 8 and two escalating doses of ISIS 2503 (4 and 6 mg/kg/day) as a 14-day continuous infusion starting on day 1. In addition, 8 patients (5 male, 3 female) received a flat dose of ISIS 2503 based on ideal body weight. Cycles were repeated every 3 weeks. Toxicities, graded according to the National Cancer Institute Common Toxicity Criteria, were recorded as maximum grade/patient for all treatment cycles. Pharmacokinetic analyses were performed to evaluate any interaction between these two agents. RESULTS: The most common nondose-limiting toxicity was hematological, manifested as neutropenia (5 grade 2, 7 grade 3, and 1 grade 4) and thrombocytopenia (10 grade 1, 5 grade 2, 5 grade 3, and 1 grade 4). Nonhematological toxicities included anorexia (7 grade 1, 3 grade 2, and 1 grade 3), nausea (10 grade 1 and 1 grade 3), fatigue (6 grade 1, 5 grade 2, and 3 grade 3), fever (6 grade 1, 2 grade 2, 1 and grade 3), and thrombosis associated with central lines (5). The plasma concentration of gemcitabine at the end of infusion was altered in the presence of ISIS 2503, leading to alterations on other pharmacokinetic parameters, but the observed differences were not clinically relevant. The plasma disposition of ISIS 2503 was not altered by gemcitabine coadministration. One partial response was documented in a heavily pretreated patient with metastatic breast cancer. Disease stabilization for greater than six cycles of treatment was observed in 5 patients. CONCLUSIONS: The combination of gemcitabine and ISIS 2503 was well tolerated and clinically active in this group of heavily pretreated patients. The recommended Phase II dose of gemcitabine (1000 mg/m(2)) and ISIS 2503 (6 mg/kg/day) warrants additional evaluation. PMID- 12538460 TI - A prospective randomized pilot study to evaluate predictors of response in serial core biopsies to single agent neoadjuvant doxorubicin or paclitaxel for patients with locally advanced breast cancer. AB - INTRODUCTION: Response to neoadjuvant chemotherapy for locally advanced breast cancer can be correlated with long-term outcomes. Surrogate end-point biomarkers may be used to assess response to the treatment. Most reported studies assessed the effects of combination chemotherapy. We assessed the feasibility of obtaining serial core breast biopsies, and correlated rates of apoptosis, proliferation, and expression of related proteins at baseline, during, and after neoadjuvant single agent chemotherapy for locally advanced breast cancer with response. EXPERIMENTAL DESIGN: Women with a histologically confirmed unresected T(3) or T(4) infiltrating carcinoma of the breast were eligible. The first 20 patients received three cycles of doxorubicin 90 mg/m(2) followed by three cycles of paclitaxel 250 mg/m(2), or the reverse. Nine women received four cycles of each (doxorubicin 60 mg/m(2) and paclitaxel 175 mg/m(2)). Cycles were administered 14 days apart with filgastrim. End points included: (a). clinical and pathological response; (b). serial apoptotic [terminal deoxynucleotidyl transferase (Tdt) mediated nick end labeling] and proliferation (immunohistochemistry, IHC) rates; and (c). expression (IHC) of estrogen receptor, HER2, bcl2, and p53. RESULTS: From April 1997 to June 2001, 29 women were randomized. Twelve patients (42%) had a clinical complete response (cCR), and 16 (55%) had a clinical partial response. Five women (17%) had a pathological complete response, 7 (24%) had microscopic residual disease, and 17 (58%) had macroscopic residual disease. Higher baseline apoptosis and proliferation were associated with an improved pathological response (P = 0.006 and 0.003, respectively). Among 14 evaluable patients, apoptosis increased in women who had a cCR to the first agent but not in women without a cCR. Estrogen receptor-positive patients had a worse pathological response (P = 0.004). CONCLUSIONS: The selected regimen is efficacious. It is feasible to obtain serial core biopsies that are informative for studies of apoptosis and IHC. This clinical design can serve as a model for combining standard chemotherapy and novel agents. PMID- 12538461 TI - Phase I study with pharmacokinetics of S-1 on an oral daily schedule for 28 days in patients with solid tumors. AB - PURPOSE: Our purpose in the study was to determine the maximum tolerated dose and dose-limiting toxicity and investigate the clinical pharmacology of S-1, a combination of tegafur, 5-chloro-2,4-dihydroxypyridine (CDHP), and potassium oxonate. EXPERIMENTAL DESIGN: Eligible patients had advanced solid tumors, adequate organ function, and no anticancer therapy in the preceding 4 weeks. Dose level 1 was 30 mg/m(2)/dose, level 2 was 40 mg/m(2)/dose, and level 3 was 35 allmg/m(2)/dose, all of the levels comprising two daily doses. S-1 was administered as a single dose at each level, and its pharmacology was studied. The first course was begun 3 days later and consisted of 28 consecutive treatment days, followed by a 1-week rest. RESULTS: Sixteen patients were enrolled; toxicity could be assessed in all of the 16 and response in 15. At dose level 1, two of nine patients developed grade 3 hyperbilirubinemia or diarrhea. Dose limiting toxicity (diarrhea) occurred in all three of the patients at dose level 2. The protocol was, therefore, amended to include an intermediate dose level (level 3), which caused grade 3 or 4 diarrhea or hyperbilirubinemia in three of four patients. Dose level 1 was thus considered as the maximum tolerated dose. Other grade 3 or 4 toxic effects at dose level 2 or 3 were granulocytopenia, nausea, and vomiting. The pharmacology of tegafur, CDHP, potassium oxonate, and fluorouracil (a metabolite of tegafur) was characterized by rapid absorption and was consistent with first-order kinetics. One patient with colorectal cancer had a durable partial response. CONCLUSIONS: The recommended S-1 dose for future studies is 30 mg/m(2) twice daily, and diarrhea is the most frequent toxic effect. Additional trials of S-1 in the treatment of patients with solid tumors are warranted. PMID- 12538462 TI - European organization for research and treatment of cancer (EORTC) 08957 phase II study of topotecan in combination with cisplatin as second-line treatment of refractory and sensitive small cell lung cancer. AB - PURPOSE: The purpose of this study was to assess the activity and toxicity of a combined regimen of topotecan and cisplatin in "sensitive" (s) and "refractory" (r) small-cell lung cancer (SCLC) patients treated previously. EXPERIMENTAL DESIGN: Patients with measurable SCLC and progressive disease after one first line regimen were eligible for the study. Patients were enrolled in two separate groups: r group (patients who failed first-line treatment <3 months from treatment discontinuation) and s group (patients who responded to first-line treatment and progressed >or=3 months after treatment discontinuation). Cisplatin was given i.v. at the dose of 60 mg/m(2) on day 1, and topotecan was administered as a daily i.v. infusion at the dose of 0.75 mg/m(2) from day 1 to 5, every 3 weeks. RESULTS: A total of 110 eligible (68 s and 42 r) patients were enrolled from 24 institutions. The main patient characteristics were as follows: median age 60 (s) and 55 (r) years, median performance status 1 for both (s) and (r). Seventy-four percent (s) and 67% (r) had extensive stage disease, including 22% and 36%, respectively, with brain metastases. A total of 398 chemotherapy courses were administered [median 4 (s) and 3 (r) per patient]. The most frequent and serious toxicity was myelosuppression. Grade IV neutropenia occurred in 62% (s) and 49% (r) of patients, with a 19% (s) and 15% (r) incidence of febrile neutropenia, and grade IV thrombocytopenia in 54% (s) and 44% (r). Most of these toxicities occurred during the first chemotherapy course and led to topotecan dose reduction and/or delay in the following courses. Grade III-IV nonhematological toxicity was uncommon. Five deaths possibly related to toxicity occurred among s patients only. Objective responses have been documented in 20 s patients, 19 partial responses and 1 complete response, (29.4% response rate; 95% confidence interval, 19-42), whereas, among r patients, 10 partial responses have been observed (23.8% response rate; 95% confidence interval, 12-39). Median survival for s and r was 6.4 and 6.1 months, respectively. CONCLUSIONS: The combination of cisplatin and topotecan, at this dose and schedule, shows activity and promising results in patients with refractory SCLC, with reversible myelosuppression being the main side effect. Additional development of this regimen, using better-tolerated schedules, is warranted in patients with refractory SCLC. PMID- 12538463 TI - Disposition of docosahexaenoic acid-paclitaxel, a novel taxane, in blood: in vitro and clinical pharmacokinetic studies. AB - PURPOSE: Docosahexaenoic acid-paclitaxel is as an inert prodrug composed of the natural fatty acid DHA covalently linked to the C2'-position of paclitaxel (M. O. Bradley et al., Clin. Cancer Res., 7: 3229-3238, 2001). Here, we examined the role of protein binding as a determinant of the pharmacokinetic behavior of DHA paclitaxel. EXPERIMENTAL DESIGN: The blood distribution of DHA-paclitaxel was studied in vitro using equilibrium dialysis and in 23 cancer patients receiving the drug as a 2-h i.v. infusion (dose, 200-1100 mg/m(2)). RESULTS: In vitro, DHA paclitaxel was found to bind extensively to human plasma (99.6 +/- 0.057%). The binding was concentration independent (P = 0.63), indicating a nonspecific, nonsaturable process. The fraction of unbound paclitaxel increased from 0.052 +/- 0.0018 to 0.055 +/- 0.0036 (relative increase, 6.25%; P = 0.011) with an increase in DHA-paclitaxel concentration (0-1000 microg/ml), suggesting weakly competitive drug displacement from protein-binding sites. The mean (+/- SD) area under the curve of unbound paclitaxel increased nonlinearly with dose from 0.089 +/- 0.029 microg.h/ml (at 660 mg/m(2)) to 0.624 +/- 0.216 microg.h/ml (at 1100 mg/m(2)), and was associated with the dose-limiting neutropenia in a maximum-effect model (R(2) = 0.624). A comparative analysis indicates that exposure to Cremophor EL and unbound paclitaxel after DHA-paclitaxel (at 1100 mg/m(2)) is similar to that achieved with paclitaxel on clinically relevant dose schedules. CONCLUSIONS: Extensive binding to plasma proteins may explain, in part, the unique pharmacokinetic profile of DHA-paclitaxel described previously with a small volume of distribution ( approximately 4 liters) and slow systemic clearance ( approximately 0.11 liters/h). PMID- 12538464 TI - Quantitative polymerase chain reaction monitoring of BCR-ABL during therapy with imatinib mesylate (STI571; gleevec) in chronic-phase chronic myelogenous leukemia. AB - PURPOSE: The purpose of our investigation was to evaluate the response and minimal residual disease by quantitative competitive PCR (QC-PCR) studies in patients with chronic myeloid leukemia (CML) treated with imatinib mesylate. EXPERIMENTAL DESIGN: One hundred eighty patients with Philadelphia chromosome (Ph)-positive chronic-phase CML after IFN-alpha failure, treated with imatinib mesylate, had 543 simultaneous cytogenetic and QC-PCR analyses at different times during their therapy. RESULTS: The median QC-PCR values [ratio-percentage of (BCR ABL/ABL transcripts) x 100] for cytogenetic response categories were: no response (Ph, >90%), 36%; minor response (Ph, 35-90%), 22%; partial response (Ph, 1-34%), 7.3%; complete response (Ph, 0%), 0.89%. There was good correlation between cytogenetic and QC-PCR studies (P < 0.001; r = 0.92) and good concordance between QC-PCR values (>10%, 2-10%, and <2%) and cytogenetic response categories (none, minor, partial, complete) with a concordance rate of 66%, and major discordance of only 10%. Of 170 samples in complete cytogenetic response, 21% still had QC PCR values of >10%, and 53% had QC-PCR values of <1%. There was excellent concordance between blood and marrow QC-PCR values (r = 0.965; P < 0.01; concordance rate, 88%; major discordance, 0%). No patient in complete cytogenetic response regardless of QC-PCR value has yet relapsed. At a median follow-up time of 26 months, higher QC-PCR values within each cytogenetic category at 3, 6, and 9 months have not been associated with a higher occurrence cytogenetic relapse or disease progression. However, the significance of this may become different with longer follow-up. CONCLUSION: QC-PCR studies provide a useful tool to monitor patients with CML on imatinib mesylate therapy. PMID- 12538465 TI - Differential expression of cancer testis genes in histological subtypes of non Hodgkin's lymphomas. AB - PURPOSE: The purpose of this study was to determine the potentialof cancer testis (CT) antigens as vaccines for non-Hodgkin's lymphomas (NHLs). EXPERIMENTAL DESIGN: Ninety-three specimens of NHLs were analyzed for their composite expression of eight CT genes (MAGE-3, MAGE-4, CT-7, HOM-MEL-40/SSX-2, SSX-1, SSX 4, HOM-TES-14/SCP-1, and HOM-TES-85). Thirty-nine of these specimens were also analyzed for their NY-ESO-1 expression. RESULTS: Only 1 of 7 cases of chronic lymphocytic leukemia expressed a CT gene (HOM-TES-14/SCP-1), and 10 follicular lymphomas were negative for all of the CT genes tested. In B-cell lymphomas, the most frequent expression of CT genes was observed in diffuse large-cell lymphomas (HOM-TES-14/SCP-1: 7 of 28; SSX-1: 5 of 28; CT-7: 2 of 28; and HOM-MEL-40/SSX-2 and HOM-TES-85: 1 of 28 positive cases). Only 1 of 8 Burkitt's and 1 of 7 lymphoblastic lymphomas expressed a CT gene (CT7 and HOM-TES-14/SCP-1, respectively). A majority (9 of 15) of T- NHLs (9 peripheral T-cell lymphomas, 2 lymphoblastic T-cell lymphomas, and 4 cases of AILD) expressed HOM-TES-14/SCP-1. CONCLUSIONS: HOM-TES-14/SCP-1, and to some degree SSX-1 and CT-7 might be candidates for lymphoma vaccine development. However, the identification of additional tumor-specific antigens with a frequent expression in lymphomas is warranted to allow for the development of widely applicable polyvalent lymphoma vaccines. PMID- 12538466 TI - Influence of adjuvant hormone therapy and chemotherapy on the immune system analysed in the bone marrow of patients with breast cancer. AB - PURPOSE: Bone marrow is a special compartment for antitumor immunological memory in patients with breastcancer. Until now, the influence of adjuvant systemic therapy on the immune system has only been investigated in peripheral blood and not in bone marrow. In this study, we analyzed the effect of hormone therapy and chemotherapy on the immune activation status in bone marrow. EXPERIMENTAL DESIGN: In 34 patients with breast cancer, bone marrow was aspirated 24 months after primary surgery and adjuvant systemic therapy. The immune system of these patients was compared with that of patients at the time of primary surgery (n = 90). Three-color flow cytometry was used to identify the number and activation state of T cells, natural killer (NK) cells, monocytes/macrophages, and subsets by means of a panel of monoclonal antibodies. RESULTS: The proportion of all T cells was significantly lower in patients after adjuvant systemic therapy than in patients with primary breast cancer or normal healthy donors. Chemotherapy apparently had a particularly suppressive effect on naive CD4 T cells and, to a lesser extent, on memory CD4 T cells. Hormone therapy apparently had a significant suppressive effect on both naive and memory CD8 T cells. The numbers of NK cells (CD56) and of monocytes/macrophages (CD14) recovered rapidly after adjuvant chemotherapy. However, subpopulations with potential antitumor reactivity, such as activated NK and NK T cells, were reduced per long term after chemotherapy. CONCLUSIONS: These findings suggest profound and long-lasting negative effects on the bone marrow immune system by present day adjuvant therapy in breast cancer. PMID- 12538468 TI - Characterization of c-kit expression in small cell lung cancer: prognostic and therapeutic implications. AB - PURPOSE: The tyrosine-kinase receptor c-kit and its ligand stem cell factor are coexpressed in many small cell lung cancer (SCLC) cell lines, leading to the hypothesis that this coexpression constitutes an autocrine growth loop. To further evaluate the frequency and pathogenic relevance of c-kit expression, tumor tissue together with the corresponding clinical data of SCLC patients was analyzed. EXPERIMENTAL DESIGN: Tumor tissue of 102 consecutive SCLC cancer patients was analyzed immunohistochemically using an affinity-purified polyclonal c-kit antibody. Immunostaining data were correlated with survival and other relevant clinical parameters. RESULTS: A positive c-kit expression was observed in 37% of patients. c-kit expression was associated with decreased survival in the likelihood-ratio-forward selection model of the Cox regression including clinically relevant risk factors (c-kit expression, age, gender, stage, tumor stage, node stage, metastasis stage, weight loss, performance status, response to chemotherapy, lactate dehydrogenase, neuronspecific enolase, hemoglobin). Only c kit expression [hazard ratio, 2.00; confidence interval (CI), 1.17-3.41; P = 0.012], response to chemotherapy (hazard ratio, 4.49; CI, 2.36-8.55; P < 0.001), and tumor stage (hazard ratio, 2.11; CI, 1.18-3.74; P = 0.008) were explanatory prognostic factors. These factors and all possible interactions between them were further analyzed in a second Cox regression model. As expected, response to chemotherapy had the highest impact on survival (hazard ratio, 3.06; CI, 1.69 5.54; P < 0.001). In patients with extensive disease, minor response to chemotherapy, and positive c-kit expression, the risk to die increased to 8.4 (hazard ratio, 2.74; CI, 1.52-4.91; P = 0.002). In a Kaplan-Meier analysis median survival of patients with minor response to chemotherapy and extensive stage was 288 days (CI, 255-321 days) when c-kit expression was negative compared with only 71 days (CI, 0-237 days) for c-kit-positive patients (log rank test: P = 0.003). CONCLUSIONS: c-kit represents a new prognostic factor in SCLC. c-kit expression is of particular clinical relevance in patients with advanced disease and poor response to chemotherapy. Given the very limited therapeutic options and unfavorable prognosis of these patients, clinical studies aimed at targeting c kit (e.g., STI571) are clearly warranted. PMID- 12538467 TI - Expression of the c-Met proto-oncogene and its possible involvement in liver invasion in adult T-cell leukemia. AB - c-Met is a tyrosine kinase receptor for hepatocyte growth factor and suggested to be involved in oncogenesis ormetastatic phenotypes in many malignancies. Adult T cell leukemia (ATL) is a neoplasia characterized by massive invasion of the leukemic cells into various organs. Recently, we have reported frequent hepatic involvement and the relationship between liver invasion and the poor prognosis in ATL. In the present study, we investigated the expression of c-Met in ATL cells and its relation to liver dysfunction. In three of four human T-cell lymphotrophic virus-I-positive T-cell lines, c-Met was expressed both at mRNA and protein levels, whereas it was not expressed in human T-cell lymphoma virus-I negative T-cell lines. The expressed c-Met should be functional, because hepatocyte growth factor could induce the autophosphorylation of c-Met. Although the viral-transactivating protein Tax has been shown to be involved in the deregulated expression of cellular genes, Tax mRNA was not detected in c-Met mRNA expressed cell lines. From freshly isolated peripheral blood mononuclear cells, the expression of c-Met mRNA was detected in 10 of 16 ATL patients but not from healthy individuals. Finally, serum transaminase levels were significantly increased in c-Met-positive ATL cases, and all of the infiltrated c-Met-positive cells into liver were shown to be multilobularly nucleated phenotype. Taken together, these data suggest for the first time that c-Met is involved in the liver invasive phenotype of ATL. PMID- 12538469 TI - Lymphocyte recovery in advanced ovarian cancer patients after high-dose chemotherapy and peripheral blood stem cell plus growth factor support: clinical implications. AB - PURPOSE: The purpose of this study was to investigate the clinical role of immunological recovery together with selected biological parameters on long-term survival in a series of ovarian cancer administered high-dose chemotherapy with peripheral blood stem cell and growth factor support. EXPERIMENTAL DESIGN: Thirty eight patients with stages IIIB-IV epithelial ovarian cancer were studied. Lymphocyte immunophenotyping for the identification of CD3(+), CD4(+), CD8(+), and CD3(-)/CD16(+)CD56(+) natural killer T cells and CD19 B cells was performed. RESULTS: Twenty-three patients (60%) had a CD3(+) cell count <850 cells/ microl. Multivariate logistic regression showed that tumor grading (chi(2) = 6.6, P = 0.010) and type of growth factor (chi(2) = 4.1, P = 0.042) retained an independent role in predicting T-cell recovery above the value of 850 cells/ microl. The 3-year time to progression (TTP) rate was 86% (95% confidence intervals, 70, 102) in cases with high CD3(+) cell count with respect to a 3-year TTP of 23% (95% confidence intervals, 8, 38) in cases with low CD3(+) cell count (P = 0.0026). The absolute number of CD3(+) cells was shown to be inversely associated with risk of progression (chi(2) = 4.8; P = 0.028), as assessed by Cox univariate analysis using CD3(+) cell count as continuous covariate. In multivariate analysis only residual tumor and status of CD3(+) cell counts retained an independent association with shorter TTP. Similar results were obtained for overall survival. CONCLUSIONS: Long-term immune reconstitution and particularly the recovery of adequate counts of CD3(+), CD4(+), and CD8(+) T cells are independent markers of longer TTP and overall survival in ovarian cancer patients receiving high-dose chemotherapy with peripheral blood stem cell and growth factor support. PMID- 12538470 TI - P7 antigen expression in human breast cancer. AB - PURPOSE: Evaluate p7 expression in human breast cancer and determine whether chemotherapy and radiation therapy effect a change in p7 expression. EXPERIMENTAL DESIGN: Using a p7-specific monoclonal antibody with immunohistochemistry and Western immunoblot analyses to assess p7 expression in archival, frozen breast cancer specimens both before and after therapy. RESULTS: A novel 7 kDa protein (p7), originally identified in multidrug-resistant ovarian and breast cancer cell lines, was found to be expressed in 21 of 64 (32%) primary, unselected human breast cancer specimens by immunohistochemistry with the use of a p7-specific monoclonal antibody, 1D7. P7 was observed in malignant cells but not in other types of cells in the breast tissue. Western blot analysis confirmed the 7 kDa polypeptide in p7-positive breast carcinomas identified by immunohistochemistry. P7 expression was significantly associated with breast cancers having distant metastasis and/or local recurrence (P = 0.027, Fisher's exact test). In addition, p7 expression was significantly increased in post-treatment breast cancer biopsy specimens compared with pretreatment breast cancer biopsy specimens in patients with locally advanced breast cancer after 5-fluorouracil chemotherapy and radiation therapy [2 of 15 (13%) pretreatment breast cancers compared with 8 of 15 (53%) post-treatment breast cancers; P = 0.016, McNemar's test]. CONCLUSIONS: These findings demonstrate that expression of p7 is associated with malignant tumor cells in primary breast cancers, especially those showing recurrent or metastatic disease. Its specific association with the malignant phenotype suggests it may have potential for novel target-based therapies. The markedly increased expression in patients with locally advanced disease after neoadjuvant therapy suggests a role for p7 in treatment outcome. PMID- 12538471 TI - Reduced expression of the small leucine-rich proteoglycans, lumican, and decorin is associated with poor outcome in node-negative invasive breast cancer. AB - PURPOSE: To examine the prognostic significance of lumican and decorin, two abundant small leucine-rich proteoglycans in breast tissue stroma. EXPERIMENTAL DESIGN: Lumican and decorin expression was examined in a cohort of 140 invasive breast carcinomas by Western blot analysis. All cases were axillary lymph node negative and treated by adjuvant endocrine therapy. RESULTS: Lumican and decorin expression was highly correlated (r = 0.45, P < 0.0001), but although low levels of lumican were associated with large tumor size (P = 0.0496), negative estrogen receptor (P = 0.0024) and progesterone receptor status (P = 0.0116), and increased host inflammatory response (P = 0.0077), low decorin levels were associated only with large tumor size (P = 0.0496). However, using univariate analysis, low levels of lumican and decorin were both associated with a shorter time to progression (P = 0.0013 and 0.0262) and poorer survival (P = 0.001 and 0.0076). In multivariate analysis using the Cox regression model, low decorin was also shown to be an independent predictive factor for recurrence (hazard ratio 2.25: 95% confidence interval 1-5, P = 0.047) and survival (hazard ratio 3.39: 95% confidence interval 1.2-9.6, P = 0.021). CONCLUSIONS: These results suggest that low levels of small leucine-rich proteoglycans in breast tumors may be associated with a worse prognosis in lymph node-negative invasive breast carcinomas and warrant further study with larger patient cohorts. PMID- 12538472 TI - Overexpression of focal adhesion kinase in primary colorectal carcinomas and colorectal liver metastases: immunohistochemistry and real-time PCR analyses. AB - PURPOSE: Focal adhesion kinase (FAK), a protein tyrosine kinase that functions in signaling events between cells and their extracellular matrix, is overexpressed in a variety of human solid tumors. To determine whether FAK expression is up regulated in colorectal cancer, we analyzed FAK mRNA and protein levels in primary colorectal tumors and colorectal liver metastases. EXPERIMENTAL DESIGN: p125(FAK) expression in formalin-fixed paraffin-embedded (FFPE) tissue was studied using immunohistochemical assays on 24 matched primary colorectal carcinomas and colorectal liver metastases as well as 18 different colorectal liver metastases using monoclonal anti-FAK 4.47. FAK mRNA expression was quantitated by real-time PCR on 39 matched normal colorectal mucosa and primary colorectal carcinomas as well as on 17 separate liver metastases. RESULTS: Elevated levels of p125(FAK) expression were demonstrated in both primary colorectal tumors and colorectal liver metastases compared with normal colorectal mucosa. Immunohistochemistry experiments demonstrated equivalent FAK expression in matched samples of colorectal primary tumors and liver metastases. Using real time PCR in 39 matched samples, FAK mRNA copy number was significantly higher in primary colorectal tumors compared with normal colorectal mucosa. FAK expression was analyzed by both real-time PCR and immunohistochemistry in a separate set of colorectal liver metastases. Immunohistochemistry demonstrated high levels of FAK expression in 89% of samples. Furthermore, FAK mRNA copies in these unmatched liver metastases were significantly higher than the primary tumor FAK mRNA copies. CONCLUSION: These experiments have shown that both primary colorectal cancers and colorectal liver metastases express high levels of FAK mRNA and p125(FAK) protein. Furthermore, the majority of colorectal liver metastases demonstrated robust FAK expression equivalent to or greater than that in the primary colorectal tumor. PMID- 12538473 TI - Quantitative O(6)-methylguanine DNA methyltransferase methylation analysis in curatively resected non-small cell lung cancer: associations with clinical outcome. AB - PURPOSE: Hypermethylation of the O(6)-methylguanine DNA methyltransferase (MGMT) promoter region leads to transcriptional repression of the MGMT gene and is a common event in primary human neoplasia. The purpose of this study was to determine the frequency and clinical relevance of MGMT gene promoter hypermethylation in curatively resected non-small cell lung cancer (NSCLC). EXPERIMENTAL DESIGN: MGMT hypermethylation, expressed as the ratio between methylated MGMT to unmethylated MYOD1 in genomic DNA, was analyzed in normal and matching tumor tissue from 90 patients with NSCLC, and a control group of 10 patients without cancer using a methylation-specific fluorogenic Real-Time PCR (Taqman) system. RESULTS: Hypermethylation of the MGMT promoter was detected in 34 of 90 (38%) tumor specimens and 16 of 90 (18%) matching normal lung tissues of patients with NSCLC, and in 0 (0%) cases of the control group without lung cancer. The mean MGMT methylation level was significantly higher in tumor than in matching normal tissue (P < 0.001). MGMT methylation in normal tissue was always accompanied with MGMT methylation in matching tumor tissue. Patients without MGMT promoter hypermethylation showed a significantly better survival than patients with MGMT promoter hypermethylation (P = 0.017). Multivariate analysis revealed MGMT promoter methylation as an independent unfavorable prognostic factor (P = 0.030). CONCLUSIONS: MGMT promoter hypermethylation is a common event in patients with primary NSCLC. This epigenetic alteration is associated with inferior survival, suggesting that MGMT promoter hypermethylation might be an important biomarker for a biological aggressive disease in NSCLC. PMID- 12538474 TI - p53 mutations in human aggressive and nonaggressive basal and squamous cell carcinomas. AB - PURPOSE: The purpose is to investigate whether aggressive basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) differ from nonaggressive BCC and SCC with respect to the p53 mutation spectrum and whether specific mutations can serve as prognostic indicators of tumor aggressiveness. EXPERIMENTAL DESIGN: We analyzed 342 tissues from patients with aggressive and nonaggressive BCCs and SCCs for p53 mutations by single-strand conformation polymorphism and nucleotide sequencing. RESULTS: p53 mutations were detected in 33 of 50 aggressive BCCs (66%), 37 of 98 nonaggressive BCCs (38%), 28 of 80 aggressive SCCs (35%), 28 of 56 nonaggressive SCCs (50%), and 3 of 29 samples of sun-exposed skin (10%). About 71% of the p53 mutations detected in aggressive and nonaggressive BCCs and SCCs were UV signature mutations. The frequency of CC to TT mutations in aggressive (36%) and nonaggressive SCCs (39%) was 2-fold higher than in aggressive (18%) and nonaggressive (14%) BCCs. In contrast, aggressive BCCs had a higher frequency (24%) of transversions than nonaggressive BCCs (8%) and aggressive (14%) and nonaggressive (11%) SCCs did. CONCLUSIONS: Our results indicate that UV radiation is responsible for the induction of p53 mutations and perhaps for the initiation of both aggressive and nonaggressive BCCs and SCCs. Although some differences in p53 mutation frequency, types of mutation, and hot spots were seen between aggressive and nonaggressive BCCs and SCCs, these factors do not constitute as clear-cut diagnostic or prognostic indicators of tumor aggressiveness. Tumor aggressiveness may be attributable to other genetic changes or events that occur during tumor progression. PMID- 12538475 TI - Detecting homozygous deletions in the CDKN2A(p16(INK4a))/ARF(p14(ARF)) gene in urinary bladder cancer using real-time quantitative PCR. AB - PURPOSE: 9p21 is a major target in the pathogenesis of human urinary bladder cancer. The locus harbors the CDKN2A/ARF tumor suppressor gene, which encodes two cell cycle regulatory proteins cyclin dependent kinase 2A (p16(INK4a)) and alternate reading frame (p14(ARF)). We have designed a real-time quantitative PCR (QPCR) application to study homozygous deletion (HD) of CDKN2A/ARF in 186 urinary bladder cancer patients. EXPERIMENTAL DESIGN: Real-time QPCR, based on simultaneous amplification of ARF and a reference gene, GAPDH, was developed and evaluated in three melanoma cell lines with HDs at the CDKN2A/ARF locus (IGR-1, SK-MEL-5, and WM-266-4). In addition, loss of heterozygosity was analyzed at the D9S942, D9S1748, and D12S99 markers. Mutation analysis of the CDKN2A/ARF gene was performed using single-strand conformational polymorphism and sequencing. Results from the present investigation were combined with previous p53 analysis of the same urinary bladder neoplasms. RESULTS: Real-time QPCR analysis showed 26 (14%) HDs, 22 (12%) hemizygous deletions, and 3 (2%) multiple duplications. Loss of heterozygosity was determined in 30 (22%) cases at the D9S942 locus, which is located between E1alpha and E1beta of the CDKN2A/ARF gene. No association was established between occurrence of genetic aberrations at 9p21 and tumor stage or grade, supporting previous suggestions that CDKN2A/ARF inactivation is an early event in bladder carcinogenesis. CONCLUSIONS: We have established a fast and efficient method for detection of HDs. Our data support the notion that inactivation, including HDs, of CDKN2A/ARF is an early event in transitional cell carcinoma. We observed separate and specific targeting of the CDKN2A and ARF genes, respectively, and that simultaneous inactivation of ARF and p53 occurs. PMID- 12538476 TI - Cell culture in esophageal squamous cell carcinoma and the association with molecular markers. AB - PURPOSE: We reported previously that the patients in whom cancer cells could be cultured as continuous cell lines had a poor prognosis of esophageal squamous cell carcinoma (ESCC) patients. In this study, to evaluate additional evidence of prognostic significance and the genetic background of cell culture, we analyzed 203 ESCC patients. EXPERIMENTAL DESIGN: Culture samples were obtained from resected 203 primary ESCC (from 1986 to 1998; R0 resection). The expression of six molecular markers was evaluated retrospectively in resected primary esophageal tumors by immunohistochemical analysis, and the capability of establishing cell lines was compared. RESULTS: Thirty-five cell lines (17.2%) were established from 203 ESCC patients: group 1 (n = 35), from whom cancer cells could be cultured as continuous cell lines, and group 2 (n = 168), from whom cell lines could not be established. The cumulative survival rate of patients in group 1 was significantly lower than that of those in group 2 (P = 0.0006). Cox's proportional hazard model revealed that cell culture capability was an independent prognostic factor (risk ratio, 1.98; P = 0.007). Univariate logistic regression analysis revealed that cell culture capability had associations with the following molecular biological factors: cyclin D1, p53, murine double minute 2, p27, and fragile histidine triad gene (P < 0.05). However, multivariate logistic regression analysis revealed that p53 protein accumulation and MDM2 protein expression predict establishment of cell line in ESCC (odds ratio, 7.72 and 8.62, respectively). CONCLUSIONS: Cell culture capability is a significant prognostic factor in ESCC. p53 and MDM2 may have a crucial role in the establishment of ESCC cell lines. PMID- 12538477 TI - Independent prognostic impact of lymphatic vessel density and presence of low grade lymphangiogenesis in cutaneous melanoma. AB - The aim of this study was to determine lymphatic vessel density (LVD) in a series of nodular melanoma and correlate the findings with the expression of several angiogenic factors, including vascular endothelial growth factor-C, basic fibroblast growth factor (bFGF), patient survival, and clinico-pathologic data. Patients with nodular melanoma and complete follow-up information were included. Lymphatic vessels were immunostained with the LYVE-1 and Podoplanin antibodies, and LVD was evaluated in both intra- and peri-tumoral (LVDpt) areas. Median LVD was 6.3 and 12.5 vessels/mm(2) in intra- and peri-tumoral areas, and coexpression of LYVE-1 and Ki-67/MIB-1 in lymphatic endothelial cells within the tumor was demonstrated, indicating active but low-grade lymphangiogenesis. Increased LVDpt was significantly associated with localization on the extremities (P = 0.005), decreased tumor thickness (P = 0.036), absence of vascular invasion (P = 0.004), brisk lymphocytic infiltration (P = 0.018), low proliferative rate by Ki-67 (P = 0.011), increased bFGF expression in tumor cells (P = 0.01) as well as in endothelial cells (P = 0.008), and decreased tumor cell expression of Ephrin-A1 (P = 0.009). Decreased LVD in intra-tumoral areas and LVDpt both predicted improved survival rates in multivariate analyses (for LVDpt, Hazard ratio: 2.1, P = 0.009). We found that decreased LVD was present in thicker and more proliferative tumors (Ki-67) and that increased LVD was significantly associated with improved patient survival in multivariate analysis. In addition, our data suggest the presence of low-grade intra-tumoral lymphangiogenesis in melanoma and a stimulating role of bFGF in lymphangiogenesis. PMID- 12538478 TI - Combined microsatellite and FGFR3 mutation analysis enables a highly sensitive detection of urothelial cell carcinoma in voided urine. AB - PURPOSE: Fibroblast growth factor receptor 3 (FGFR3) mutations were reported recently at a high frequency in low-grade urothelial cell carcinoma (UCC). We investigated the feasibility of combining microsatellite analysis (MA) and the FGFR3 status for the detection of UCC in voided urine. EXPERIMENTAL DESIGN: In a prospective setting, 59 UCC tissues and matched urine samples were obtained, and subjected to MA (23 markers) and FGFR3 mutation analysis (exons 7, 10, and 15). In each case, a clinical record with tumor and urine features was provided. Fifteen patients with a negative cystoscopy during follow-up served as controls. RESULTS: A mutation in the FGFR3 gene was found in 26 (44%) UCCs of which 22 concerned solitary pTaG1/2 lesions. These mutations were absent in the 15 G3 tumors. For the 6 cases with leukocyturia, 46 microsatellite alterations were found in the tumor. Only 1 of these was also detected in the urine. This was 125 of 357 for the 53 cases without leukocyte contamination. The sensitivity of MA on voided urine was lower for FGFR3-positive UCC (15 of 21; 71%) as compared with FGFR3 wild-type UCC (29 of 32; 91%). By including the FGFR3 mutation, the sensitivity of molecular cytology increased to 89% and was superior to the sensitivity of morphological cytology (25%) for every clinical subdivision. The specificity was 14 of 15 (93%) for the two (molecular and morphological) cytological approaches. CONCLUSIONS: Molecular urine cytology by MA and FGFR3 mutation analysis enables a highly sensitive and specific detection of UCC. The similarity of molecular profiles in tumor and urine corroborate their clonal relation. PMID- 12538479 TI - Expression of endocrine gland-derived vascular endothelial growth factor in ovarian carcinoma. AB - The first tissue-specific angiogenic molecule, endocrine gland-derived vascular endothelial growth factor (EG-VEGF), was identified recently in human ovary, raising hopes of developing tumor type-specific angiogenesis inhibitors. In the present study, we analyzed the expression of EG-VEGF mRNA in normal human tissues and ovarian neoplasms by quantitative real-time reverse transcription-PCR. EG VEGF mRNA was expressed in all ovarian neoplasms examined. No significant difference was identified among benign, low malignant potential neoplasms or stage I ovarian cancer, all of which exhibited 2-fold lower mRNA levels compared with normal premenopausal ovaries. EG-VEGF mRNA levels further decreased in late stage compared with early stage carcinomas (P < 0.05) and were consistently lower in laser capture microdissected tumor islets compared with surrounding stroma. EG VEGF was undetectable by reverse transcription-PCR in 17 established epithelial ovarian cancer cell lines or in cultured human ovarian surface epithelial cells, whereas it was detected in peripheral blood as well as tumor-infiltrating T lymphocytes. Finally, in contrast to VEGF, EG-VEGF mRNA levels did not correlate with clinical outcome in advanced ovarian carcinoma. These results suggest that EG-VEGF is most likely derived from nonepithelial components of ovarian carcinomas and may play a marginal role in promoting angiogenesis in advanced ovarian carcinoma. We postulate that EG-VEGF-targeted antiangiogenic therapy may prove useful in early stage but not in advanced stage ovarian carcinoma. PMID- 12538480 TI - Hepatocyte growth factor and c-Met in cervical intraepithelial neoplasia: overexpression of proteins associated with oncogenic human papillomavirus and human immunodeficiency virus. AB - PURPOSE: High prevalence of squamous cervical intraepithelial neoplasia (CIN) linked to oncogenic human papillomavirus (HPV) exits in HIV-infected women. Hepatocyte growth factor (HGF) and its receptor, c-Met, promote cell proliferation and are involved in tumor progression. Nothing is yet known about their expression in low- and high-grade CIN. Therefore, the expression, localization, and behavior of HGF and c-Met in normal and dysplastic cervical epithelium were investigated. EXPERIMENTAL DESIGN: We studied normal cervical mucosa from 10 healthy women, and low- and high-grade cervical lesions, uninfected (condyloma acuminata) or infected with oncogenic HPVs, from 40 HIV negative and 48 HIV-positive women, using in situ molecular techniques, immunocytochemistry and morphoquantitative methods. RESULTS: In 154 oncogenic HPV infected CIN encountered in biopsy samples, the total number of epithelial cell layers increased significantly during lesion progression. This number was significantly higher in HIV-positive than in HIV-negative women for CIN1 and CIN2 (P < 0.025 to P < 0.01). In HIV-negative women, the number and percentage of HGF and c-Met immunostained cell layers, and the intensity of immunostaining were enhanced in oncogenic HPV-infected lesions as compared with normal mucosa and condyloma acuminata. The latter parameters were significantly higher in tissues of HIV-positive women (oncogenic HPV-infected CIN1 and CIN2, normal-appearing mucosa contiguous to CIN, condyloma acuminata) than in the corresponding tissues of HIV-negative women (P < 0.025 to P < 0.0001). CONCLUSIONS: Overexpression of HGF/c-Met complex strongly correlates with oncogenic HPV and HIV infection. This overexpressed complex may stimulate cell proliferation in condyloma acuminata and participate in tumor progression in oncogenic HPV-infected lesions. PMID- 12538481 TI - Effective combination of chemotherapy and dendritic cell administration for the treatment of advanced-stage experimental breast cancer. AB - PURPOSE: The goal of this study was to investigate the utility of a new approach to the treatment of advanced stage breast cancer, a combination of chemotherapy and dendritic cell (DC) administration. EXPERIMENTAL DESIGN: Mice bearing mammary adenocarcinoma expressing a model tumor antigen, influenza virus HA (DA3-HA), and parental tumor (DA3) were treated with different doses of paclitaxel with or without DCs. Paclitaxel was injected three times weekly, DCs were injected either i.v. or into tumor site (t.s.) 36 h after each injection of paclitaxel. Apoptosis was measured using Annexin V binding or terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL) assays. CD8-mediated response of T cells to HA derived peptide epitope was measured in an enzyme-linked immunospot (ELISPOT) assay. CD4-mediated response of T cells to HA-derived peptide was measured in proliferation assay. Nonspecific T-cell proliferation was measured in response to ConA and immobilized anti-CD3 and anti-CD28 antibodies. RESULTS: We have selected the dose of paclitaxel that induced a substantial level of apoptosis and moderate inhibition of T-cell function. Combined treatment resulted in the induction of HA specific CD8-mediated response in all nine of the tested mice, and CD4-mediated responses in four of six treated mice. These effects were observed only if DCs were injected into tumor site, but not when injected i.v. No specific responses were found in mice treated with either chemotherapy or DCs alone. Injection of dexamethasone together with paclitaxel did not affect the induction of immune responses. Significant antitumor effect of combined treatment was observed in DA3 HA tumor-bearing mice as well as in mice bearing parental DA3 tumor. CONCLUSION: The combination of DC administration with repeated cycles of chemotherapy and dexamethasone (conditions similar to real clinical practice) resulted in the induction of antitumor response despite the immunosuppression induced by such treatment. PMID- 12538482 TI - Zoledronic acid exhibits inhibitory effects on osteoblastic and osteolytic metastases of prostate cancer. AB - PURPOSE: In this study we have examined the effects of zoledronic acid (ZA), a new-generation bisphosphonate, on prostate cancer (CaP) cells in vitro, and on both osteoblastic and osteolytic CaP metastases in animal models. EXPERIMENTAL DESIGN: In vitro, CaP cells were treated with ZA, and the effects on proliferation, cell cycle, and apoptosis were determined. In vivo, PC-3, and LuCaP 23.1 s.c. and tibial tumors were treated with ZA. Effects on bone and tumor were determined by histomorphometry and immunohistochemistry. RESULTS: ZA decreased proliferation of CaP cells, and caused G(1) arrest and apoptosis of CaP cells in vitro. In vivo, s.c. CaP tumor growth was not affected by ZA. However, growth of osteoblastic and osteolytic metastases of CaP was inhibited significantly in vivo. Matrix metalloproteinase-2, matrix metalloproteinase-9, and Cathepsin K levels were decreased in osteolytic bone metastases after ZA administration. CONCLUSIONS: In conclusion, we have shown that ZA has significant antitumor effects on CaP cells in vitro and in vivo. Antiosteolytic activity and the antitumor effects of this compound could benefit CaP patients with bone metastases. PMID- 12538483 TI - Timed sequential therapy of acute leukemia with flavopiridol: in vitro model for a phase I clinical trial. AB - PURPOSE: The survival of adults with acute leukemias remains unsatisfactory and requires new treatment approaches. Flavopiridol modulates cell cycle progression, inhibits transcription, and induces apoptosis. We designed an in vitro model of timed sequential therapy for acute leukemia to determine whether flavopiridol can: (a). trigger apoptosis in fresh acute leukemia; and (b). recruit surviving leukemic cells to a proliferative state, thereby priming such cells for the S phase-related cytotoxicity of 1-beta-D-arabinofuranosylcytosine (ara-C). EXPERIMENTAL DESIGN: Bone marrow cells from 20 adults with relapsed and refractory acute leukemias were enriched for blasts by Ficoll Hypaque sedimentation. Blasts were cultured on day 0 in flavopiridol 250 nM for 24 h, removed from flavopiridol for 24 h, and then cultured in ara-C 1 microM for an additional 72 h (F(250)A(1)). Apoptosis and cell cycle phase distribution were estimated from cells stained with propidium iodide. Cell survival was determined after the 72 h ara-C exposure by double cytofluorescence assay with fluorescein diacetate and propidium iodide. RESULTS: Flavopiridol induced a 4.3-fold increase in apoptosis in human leukemia samples within the first 24 h of culture. Subsequent removal of flavopiridol led to a 1.7-fold increase in the proportion of cells in S phase by day 2. Mean survival in F(250)A(1) cultures after 72 h exposure to ara-C was 35.6% compared with flavopiridol alone (F(250)A(0), 56.1%; P = 0.0003) and ara-C alone (F(0)A(1), 65.2%; P < 0.00001). CONCLUSIONS: Flavopiridol induces apoptosis in marrow blasts from patients with refractory acute leukemias. Furthermore, flavopiridol pretreatment increases the proapoptotic and cytotoxic effects of ara-C. The advantage of sequential FP(250)A(1) over either agent alone is seen for both acute myelogenous leukemia and acute lymphoblastic leukemia. These findings support a clinical trial of timed sequential therapy where flavopiridol is given for cytoreduction and subsequent priming of remaining leukemic cells for enhanced cycle-dependent drug cytotoxicity. PMID- 12538484 TI - Inhibition of constitutive STAT3 activity sensitizes resistant non-Hodgkin's lymphoma and multiple myeloma to chemotherapeutic drug-mediated apoptosis. AB - Hematopoietic malignancies have been shown to depend on cytokine growth factor autocrine/paracrine loops for growth and differentiation. This results in the constitutive activation of cytokine-mediated transcription factors like signal transducer and activators of transcription (STAT) 3 in non-Hodgkin's lymphoma (NHL) and multiple myeloma (MM). Recent evidence demonstrates that cytokines also contribute to a drug-resistant phenotype in many tumor cell types. We hypothesized that inhibitors of the STAT3 pathway would sensitize drug-resistant and endogenous cytokine-dependent NHL and MM tumor cells to the cytotoxic effects of chemotherapeutic drugs. We examined an AIDS-related NHL cell line, 2F7, known to be dependent on interleukin (IL)-10 for survival and an MM cell line, U266, known to be dependent on IL-6 for survival. IL-10 and IL-6 signal the cells through the activation of Janus kinase (JAK)1 and JAK2, respectively. Thus, we investigated the effect of two chemical STAT3 pathway inhibitors, namely, piceatannol (JAK1/STAT3 inhibitor) and tyrphostin AG490 (JAK2/STAT3 inhibitor), on the tumor cells for sensitization to therapeutic drugs. We demonstrate by phosphoprotein immunoblotting analysis and electrophoretic mobility shift analysis that piceatannol and AG490 inhibit the constitutive activity of STAT3 in 2F7 and U266, respectively. Furthermore, piceatannol and AG490 sensitize 2F7 and U266 cells, respectively, to apoptosis by a range of therapeutic drugs including cisplatin, fludarabine, Adriamycin, and vinblastine. The specificity of the inhibitors was corroborated in experiments showing that piceatannol had no effect on U266 and, likewise, AG490 has no effect on 2F7. The sensitization observed by these inhibitors correlated with the inhibition of Bcl-2 expression in 2F7 and Bcl-xL expression in U266. Altogether, these results demonstrate that STAT3 pathway inhibitors are a novel class of chemotherapeutic sensitizing agents capable of reversing the drug-resistant phenotype of cytokine-dependent tumor cells. PMID- 12538485 TI - In vivo antitumor activity of SU11248, a novel tyrosine kinase inhibitor targeting vascular endothelial growth factor and platelet-derived growth factor receptors: determination of a pharmacokinetic/pharmacodynamic relationship. AB - One challenging aspect in the clinical development of molecularly targeted therapies, which represent a new and promising approach to treating cancers, has been the identification of a biologically active dose rather than a maximum tolerated dose. The goal of the present study was to identify a pharmacokinetic/pharmacodynamic relationship in preclinical models that could be used to help guide selection of a clinical dose. SU11248, a novel small molecule receptor tyrosine kinase inhibitor with direct antitumor as well as antiangiogenic activity via targeting the vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), KIT, and FLT3 receptor tyrosine kinases, was used as the pharmacological agent in these studies. In mouse xenograft models, SU11248 exhibited broad and potent antitumor activity causing regression, growth arrest, or substantially reduced growth of various established xenografts derived from human or rat tumor cell lines. To predict the target SU11248 exposure required to achieve antitumor activity in mouse xenograft models, we directly measured target phosphorylation in tumor xenografts before and after SU11248 treatment and correlated this with plasma inhibitor levels. In target modulation studies in vivo, SU11248 selectively inhibited Flk-1/KDR (VEGF receptor 2) and PDGF receptor beta phosphorylation (in a time- and dose-dependent manner) when plasma concentrations of inhibitor reached or exceeded 50-100 ng/ml. Similar results were obtained in a functional assay of VEGF-induced vascular permeability in vivo. Constant inhibition of VEGFR2 and PDGF receptor beta phosphorylation was not required for efficacy; at highly efficacious doses, inhibition was sustained for 12 h of a 24-h dosing interval. The pharmacokinetic/pharmacodynamic relationship established for SU11248 in these preclinical studies has aided in the design, selection, and evaluation of dosing regimens being tested in human trials. PMID- 12538486 TI - Hepatitis B virus X protein sensitizes hepatocellular carcinoma cells to cytolysis induced by E1B-deleted adenovirus through the disruption of p53 function. AB - Replication-selective adenovirus has been reported to kill tumor cells and hold promise for cancer therapy. In this study, we constructed an E1B M(r) 55000 deleted adenovirus, designated Ad5WS1, and examined its cytolytic effect on human hepatocellular carcinoma (HCC) cell lines with various p53 status. The results show that Ad5WS1 lysed HCC cells lacking p53 transcription activity. However, this effect was not observed in cells harboring functional p53. Because loss of p53 transcription activity can be induced by binding to hepatitis B virus X protein (HBx), we generated HBx stable transfectants from Chang liver cells and examined their susceptibility to Ad5WS1-induced cytolysis. Expression of HBx in Chang liver cells changed the location of p53 from the nucleus to the cytoplasm, which mostly coincided with the location of HBx in the cytoplasm. Disruption of p53 transcription activity by HBx in Chang liver cells rendered them susceptible to infection with Ad5WS1. Furthermore, Ad5WS1 exerted antitumor effect, especially when combined with chemotherapeutic agent cisplatin, in BALB/c mice bearing HBx-expressing HCC. Our results suggest that E1B M(r) 55000-deleted adenovirus may have therapeutic potential for the treatment of HCC with loss of p53 transcription activity or with HBx expression. PMID- 12538487 TI - Function of nuclear factor kappaB in pancreatic cancer metastasis. AB - PURPOSE: We seek to elucidate the role of constitutive nuclear factor kappaB (NFkappaB) activity in human pancreatic cancer cells. We have demonstrated that the transcription factor NFkappaB is activated constitutively in human pancreatic adenocarcinoma and human pancreatic cancer cell lines but not in normal pancreatic tissues or in immortalized/nontumorigenic pancreatic epithelial cells, suggesting that NFkappaB plays a critical role in development of pancreatic adenocarcinoma. EXPERIMENTAL DESIGN: By pooling all of the puromycin resistant clones after inhibitor of nuclear factor-kappaB phosphorylation mutant (IkappaBalphaM) retroviral infection, we generated pancreatic tumor cell lines that express a IkappaBalphaM (S32, 36A) that blocks NFkappaB activity. Inhibition of metastatic phenotype was assayed in an orthotopic nude mouse model. NFkappaB activity was determined by electrophoretic mobility shift assay, and the expression of NFkappaB downstream target genes was analyzed by Northern, Western, and immunohistochemical analyses. RESULTS: We showed that inhibiting constitutive NFkappaB activity by expressing IkappaBalphaM suppresses liver metastasis, but not tumorigenesis, from the metastatic human pancreatic tumor cell line AsPc-1 in an orthotopic nude mouse model. Furthermore, inhibiting NFkappaB activation by expressing IkappaBalphaM significantly reduced in vivo expression of a major proangiogenic molecule, vascular endothelial growth factor, and, hence, decreased neoplastic angiogenesis. Inhibiting NFkappaB activation by expressing IkappaBalphaM and using pharmacologic NFkappaB inhibitor PS-341 also significantly reduced cytokine-induced vascular endothelial growth factor and interleukin-8 expression in AsPc-1 pancreatic cancer cells. CONCLUSION: These results demonstrated that the inhibition of NFkappaB signaling can suppress the angiogenic potential and metastasis of pancreatic cancer, and suggest that the NFkappaB signaling pathway is a potential target for anticancer agents. PMID- 12538488 TI - Reduction of vascular and permeable regions in solid tumors detected by macromolecular contrast magnetic resonance imaging after treatment with antiangiogenic agent TNP-470. AB - PURPOSE: The availability of noninvasive techniques to detect the effects of antiangiogenic agents is critically important for optimizing treatment of cancer with these agents. Magnetic resonance imaging (MRI) is one such noninvasive technique that is routinely used clinically. EXPERIMENTAL DESIGN: In this study, we have evaluated the use of MRI of the intravascular contrast agent albumin GdDTPA to detect the effects of the antiangiogenic agent TNP-470 on the vascular volume and permeability of the MatLyLu prostate cancer model. RESULTS: TNP-470 treated tumors demonstrated a significant decrease of vascular volume, as well as a significant reduction in vascular and permeable regions, compared with volume matched control tumors. Although the fractional volume of permeable regions in the tumor decreased, the average value of tumor permeability did not decrease significantly. This was attributable to increase in permeability in some regions of the tumor. These regions were mostly associated with low vascular volume. ELISA assays of control and treated MatLyLu tumors also detected a significant increase of vascular endothelial growth factor in the TNP-470-treated tumors. CONCLUSION: MRI detected significant changes in tumor vascular characteristics after treatment with TNP-470. PMID- 12538489 TI - Effect of dimethyl sulfoxide on bladder tissue penetration of intravesical paclitaxel. AB - Our laboratory has shown that the efficacy of bladder cancer intravesical therapy is in part limited by the poor penetration of drugs into the urothelium. We further found that paclitaxel, because of its lipophilicity, shows a higher penetration than other commonly used drugs such as mitomycin C and doxorubicin. However, the commercial formulation of paclitaxel (i.e., Taxol) contains Cremophor, which forms micelles that entrap the drug and reduce its free fraction. The present study evaluated the effect of DMSO on paclitaxel release from Cremophor micelles and paclitaxel penetration in bladders of dogs given an intravesical dose of paclitaxel (500 microg/20 ml in 0.22% Cremophor, 0.21% ethanol, and 50% DMSO). Cremophor produced a concentration-dependent reduction of the free fraction of paclitaxel (reduced to 23% at 0.25% Cremophor). This Cremophor effect was reversed by DMSO in a concentration-dependent manner, resulting in a 92% free fraction at 50% DMSO. DMSO also increased the average size of Cremophor micelles from 13 nm to 230 nm at 50% DMSO. A comparison of the tissue penetration data in the presence of Cremophor and/or DMSO indicates the following effects of DMSO: (a). increase in urine production rate and, consequently, a 36% reduction of the final urine concentration; (b). 2-fold increase in paclitaxel penetration across bladder urothelium; (c). increase in drug removal from bladder tissues (30% more rapidly); and (d). a 60% increase of the amount of drug in bladder tissue. These results indicate that DMSO caused rearrangement of Cremophor micelles, reversed the entrapment of paclitaxel in Cremophor micelles and thereby increased the free fraction of paclitaxel in solution, enhanced the urine production rate and enhanced drug removal by the perfusing capillaries, with an overall effect of increasing the bladder tissue delivery of paclitaxel formulated in Cremophor. PMID- 12538490 TI - Interleukin-6 promotes androgen-independent growth in LNCaP human prostate cancer cells. AB - PURPOSE: Prostate cancer frequently progresses from an initial androgen dependence to androgen independence, rendering the only effective androgen ablation therapy useless. The mechanism underlying the androgen-independent progression is incompletely understood. Interleukin (IL)-6 has been implicated in this androgen-independent progression. In this study, we tested whether IL-6 induces androgen-independent growth both in vitro and in vivo. EXPERIMENTAL DESIGN: IL-6 was expressed in androgen-sensitive LNCaP cells. The effects of IL-6 on androgen receptor activity was determined by Northern blots and gel shift assays. The effects of IL-6 on LNCaP cell growth were determined in vitro by MTT assay and in vivo. RESULTS: IL-6 can enhance the growth of androgen-sensitive LNCaP cells in the androgen-deprived condition in vitro, which is accompanied by elevation of androgen-regulated prostate-specific antigen mRNA expression. IL-6 promotes androgen-sensitive LNCaP cell tumor growth in the castrated male mice. IL-6 enhances androgen receptor DNA binding activity and nuclear translocation. The androgen-independent phenotype induced by IL-6 in LNCaP cells is accompanied by significant activation of signal transducers and activators of transcription 3 and mitogen-activated protein kinase signal pathways. CONCLUSIONS: These studies clearly provide experimental evidence that IL-6 initiates and/or enhances the transition of prostate cancer cells from an androgen-dependent to an androgen independent phenotype. PMID- 12538491 TI - Continuous infusion of endostatin inhibits differentiation, mobilization, and clonogenic potential of endothelial cell progenitors. AB - PURPOSE: We investigated the effect of endostatin on differentiation, mobilization, and clonogenic potential of circulating endothelial cell (EC) progenitors, and whether the effect of endostatin was improved by continuous infusion (CI) versus bolus administration. EXPERIMENTAL DESIGN: Four-color flow cytometry and clonogenic EC cultures were used to study EC progenitors in tumor free mice, tumor-bearing immunodeficient mice, and immunodeficient mice xenotransplanted with human bone marrow (BM) cells. RESULTS: Endostatin significantly reduced the number of circulating EC progenitors in tumor-free BALB/c mice. The effect of endostatin on EC progenitors was enhanced significantly in mice treated with CI drug treatment. When immunodeficient mice xenotransplanted with human BM cells were treated with CI of endostatin we observed a significant decrease in the engraftment and differentiation of human BM-derived EC progenitors. Numbers of circulating EC progenitors increased 7-fold in immunodeficient mice bearing human lymphoma. In this preclinical model, treatment with CI of endostatin inhibited host murine EC progenitor mobilization and human tumor growth. Furthermore, the clonogenic potential of EC progenitors was impaired severely. CONCLUSIONS: Endostatin is a potent inhibitor of the mobilization and clonogenic potential of human and murine EC progenitors, and its preclinical activity is increased significantly in CI compared with bolus administration. These observations might be useful in the design of future clinical trials. PMID- 12538492 TI - A novel mechanism for aspirin-mediated growth inhibition of human colon cancer cells. AB - PURPOSE: The molecular mechanisms by which aspirin and other nonsteroidal anti inflammatory drugs exert chemopreventative effects in colon cancer are unclear and complex. Current investigations focus on the chemopreventive properties of nonsteroidal anti-inflammatory drugs, independent of their ability to inhibit cyclooxygenase (COX) activity, and presumably, identification of non-COX pathways will suggest new targets for clinical use. It was demonstrated recently that aspirin results in reduced microsatellite instability in colorectal cancer cells. We hypothesized that aspirin treatment might alter expression of DNA mismatch repair (MMR) proteins, representing another potential non-COX mechanism for its action. EXPERIMENTAL DESIGN: In this study, we have examined the effects of aspirin on the cellular growth rates, MMR protein levels, cell cycle analysis and apoptosis in MMR-deficient (HCT116) and MMR-proficient (HCT116+chr3 and SW480) human colon cancer cell lines. RESULTS: We found that treatment with aspirin inhibited the growth of these three cancer cell lines. In HCT116+chr3 cells, treatment with 1 mM of aspirin increased expression of the hMLH1 and hPMS2 proteins by 2.5-fold and 2-fold, respectively, and increased expression of the hMSH2 and hMSH6 proteins by 2-3-fold. For SW480 cells, treatment with 1 and 5 mM of aspirin increased expression of the hMLH1 and hPMS2 proteins by 2-4-fold and 3 5-fold, respectively, and increased expression of the hMSH2 and hMSH6 proteins by 3-7-fold. For all three of the cell lines, treatment with 1 and 2.5 mM of aspirin induced apoptosis at 48 and 72 h. Aspirin induced G(0)/G(1) cell cycle arrest in HCT116 cells. CONCLUSIONS: We conclude that aspirin acts through COX-independent mechanisms by resulting in an increase in MMR protein expression and subsequent apoptosis, which might serve as an additional means of growth inhibition in aspirin-treated human colon cancer cells. PMID- 12538493 TI - C-Jun NH(2)-terminal kinase mediates proliferation and tumor growth of human prostate carcinoma. AB - PURPOSE: C-Jun NH(2)-terminal kinase (JNK) has been implicated in numerous functions including stress responses, apoptosis,and transformation. The role in transformation is based largely on studies of isolated cell types with little indication of whether JNK plays a general role in a specific human tumor type or whether this occurs in vivo. EXPERIMENTAL DESIGN: We examined 9 human prostate carcinoma cell lines in vitro and a representative line in vivo. RESULTS: For all of the cell lines proliferation is highly correlated with serum-supported JNK activity (r(Pearson) = 0.91; P = 0.004), whereas no relationship was observed for 10 human breast cancer cell lines (r(Pearson) = -0.32). Treatment with characterized antisense oligonucleotides complementary to sequences common to either the JNK1 or JNK2 family of isoforms showed that, whereas antisense JNK1 inhibited growth by a maximum of 57%, antisense JNK2 inhibited proliferation up to 80%. Sense and scrambled control oligonucleotides had little effect (average 3.7 +/- 1.5%). Moreover, systemic treatment of mice bearing established xenografts of PC3 prostate carcinoma cells with antisense JNK1 and JNK2 led to inhibition tumor growth by 57% (P < 0.002) and 80% (P < 0.001), respectively. The difference is significant (P < 0.012). Combined antisense treatment led to a significant increase in frequency of tumor regression (P = 0.022). CONCLUSION: These results indicate that JNK is required for growth of prostate carcinoma cells in vitro and in vivo, and additionally indicate that JNK2 plays a dominant role. The JNK pathway is a novel target in the treatment of prostate carcinoma. PMID- 12538494 TI - Examination of the antiproliferative activity of iron chelators: multiple cellular targets and the different mechanism of action of triapine compared with desferrioxamine and the potent pyridoxal isonicotinoyl hydrazone analogue 311. AB - PURPOSE: Tumors are sensitive to iron (Fe) chelation therapy with the clinically used chelator desferrioxamine (DFO). Recently, the potent inhibitor of ribonucleotide reductase, Triapine, has entered clinical trials as an anticancer agent. This compound is a potential Fe chelator, but despite this, no investigations have examined its effect on cellular Fe metabolism. This is essential for understanding its mechanism of action and clinical effects. EXPERIMENTAL DESIGN: We compared the effect of Triapine with DFO, and also with the novel Fe chelator, 311, which shows marked antiproliferative activity. This latter ligand was relevant to compare, because it is tridentate like Triapine and shares structural similarity. We assessed the effects of chelators on proliferation, Fe uptake, Fe efflux, the expression of cell cycle control molecules, and iron-regulatory protein-RNA-binding activity. Redox activity was determined by ascorbate oxidation, benzoate hydroxylation, plasmid DNA degradation, and the precipitation of cellular DNA. These studies have been performed using several neuroepithelioma and neuroblastoma cell lines and a variety of normal cell types including fibroblasts, umbilical vein endothelial cells, skeletal muscle cells, monocyte-derived macrophages, and bone marrow stem cells. RESULTS: Triapine was twice as effective as DFO at mobilizing (59)Fe from prelabeled cells but was much less efficient than 311. In terms of preventing (59)Fe uptake from Tf, Triapine and DFO had similar activity, having far less efficacy than 311. All three of the chelators showed greater activity against the proliferation of neoplastic than of normal cells, the effect of 311 and Triapine being similar and these two chelators being significantly (P < 0.0001) more active than DFO. Complexation of Triapine with Fe had no appreciable effect on its antiproliferative activity, whereas addition of Fe totally inhibited the effects of DFO and 311. Furthermore, the Triapine Fe complex was shown to be redox active. CONCLUSION: The cytotoxic mechanism of action of Triapine was different from that of DFO and 311, with the combined action of Fe chelation and free radical generation being involved. PMID- 12538495 TI - Possible role of ceramide as an indicator of chemoresistance: decrease of the ceramide content via activation of glucosylceramide synthase and sphingomyelin synthase in chemoresistant leukemia. AB - We investigated the possibility of the proapoptotic lipid ceramide as an indicator of chemoresistance in leukemia. Doxorubicin (DOX) increased the ceramide level and apoptosis in drug-sensitive HL-60 cells but not in drug resistant HL-60/ADR cells, under the condition that the uptake of DOX was not different between the two cell lines. In addition, exogenous N-acetylsphingosine (C2-ceramide) enhanced DOX-induced apoptosis in HL-60/ADR cells without affecting the expression of multidrug resistant-1 protein (MDR 1) and the uptake of DOX. A lower level of ceramide with higher activities of glucosylceramide synthase (GCS) and sphingomyelin synthase (SMS) was detected in HL-60/ADR cells than in HL-60 cells. In contrast, HL-60/GCS cells, overexpressing GCS, significantly inhibited DOX-induced ceramide increase and apoptosis. These observations suggest the involvement of ceramide regulation in drug resistance of leukemia cells. In vivo, the level of ceramide was lower in chemoresistant leukemia patients (6.4 +/- 1.8 pmol/nmol phosphate; n = 14) than in chemosensitive patients (9.5 +/- 2.7 pmol/nmol phosphate; n = 9), and the activities of GCS and SMS were more than 2 fold higher in chemoresistant leukemia cells than in chemosensitive cells. MDR-1 protein was faintly expressed in one of four chemoresistant patients, but Bcl-2 were clearly detected in four patients. Therefore, it is suggested that a decrease of the ceramide level via activation of GCS and SMS is associated with the chemoresistant condition in leukemia, probably in relation to Bcl-2 but not to MDR-1 expression. PMID- 12538496 TI - Elevated sod2 activity augments matrix metalloproteinase expression: evidence for the involvement of endogenous hydrogen peroxide in regulating metastasis. AB - PURPOSE: Elevated manganese superoxide dismutase (Sod2) levels have been reported to be associated with an increased frequency of tumor invasion and metastasis in certain cancers, and the aim of this study is to examine the molecular mechanisms by which this occurs. EXPERIMENTAL DESIGN: Sod2 and catalase overexpressing HT 1080 fibrosarcoma cell lines were used to evaluate the H(2)O(2)-dependent regulation of matrix metalloproteinase (MMP)-1 promoter activity, mitogen activated protein (MAP) kinase signaling, DNA-binding activity, and MMP mRNA levels. The invasive and metastatic potential of Sod2 overexpressing cells was characterized using subrenal capsular implantation or tail vein injection of tumor cells into nude mice, respectively. RESULTS: Our data reveal that Sod2 overexpression increases the DNA-binding activity of transcription factors critical for MMP expression but also enhances MMP-1 promoter activity via the Ras//MAP/extracellular signal-regulated kinase (MEK) signaling cascade. A single nucleotide polymorphism that creates an Ets site at position -1607 bp confers Sod2-dependent MMP-1 promoter activity. Sod2 overexpression also increases the mRNA levels of MMPs-2, -3, -7, -10, -9, -11 and enhances the metastatic potential of fibrosarcoma cells when implanted in immunodeficient mice. The Sod2-dependent increases in AP-1 and SP-1 DNA-binding activity, MMP-1 promoter activity, general MMP expression, and collagen degradation can be reversed by the hydrogen peroxide detoxifying enzyme, catalase. CONCLUSION: MMPs play a critical role in the process of stromal invasion and metastasis, and these findings suggest that the association between increased Sod2 and poor prognosis in certain cancers may be attributed to elevated MMP production. PMID- 12538497 TI - Gastric epithelial reactive oxygen species prevent normoxic degradation of hypoxia-inducible factor-1alpha in gastric cancer cells. AB - The expression of hypoxia inducible factor (HIF)-1alpha protein is tightly regulated by cellular oxygen status. Namely, HIF-1alpha protein is degraded rapidly in normoxic cells, whereas hypoxia stabilizes HIF-1alpha to transactivate hypoxia-responsive genes. Here we show that HIF-1alpha protein is expressed aberrantly in gastric cancer cells under normoxia in a reactive oxygen species (ROS)-dependent manner. The normoxic expression of HIF-1alpha in concordance with its DNA binding activity enhances the transcription of target genes such as vascular endothelial growth factor. The aberrant normoxic expression of HIF 1alpha is not associated with genetic abnormalities such as the loss of von Hippel-Lindau tumor suppressor, but is well correlated with endogenous ROS (hydrogen peroxide) generation. HIF-1alpha expression is blocked by nonmitochondrial ROS inhibitors, but not by inhibitors of mitochondrial electron transfer, which indicates that nonmitochondrial ROS stabilize HIF-1alpha protein in these cells. Gastric epithelial ROS have been linked to Helicobacter pylori induced gastric carcinogenesis. This study demonstrates for the first time that ROS from H. pylori-infected gastric epithelial cells induce HIF-1alpha expression and subsequently activate HIF-1alpha-mediated transcription. Taken together, these results provide a novel mechanism of HIF-1alpha stabilization in gastric cancer, and demonstrate that gastric epithelial ROS, endogenously generated or H. pylori-stimulated, lead to the constant expression of HIF-1alpha protein under normoxia. PMID- 12538498 TI - Second International Conference on Recent Advances and Future Directions in Endocrine Manipulation of Breast Cancer: summary consensus statement. PMID- 12538500 TI - Predictions from a preclinical model: studies of aromatase inhibitors and antiestrogens. AB - We have developed a tumor model for studying the effects of aromatase inhibitors and antiestrogens in vivo. The model simulates postmenopausal breast cancer patients with estrogen-dependent tumors. This model utilizes human estrogen dependent breast cancer cells transfected with the aromatase gene (MCF-7(CA)), which are inoculated in Matrigel s.c. into ovariectomized nude mice. The estrogen receptor-positive cells produce sufficient estrogen by aromatization to stimulate their proliferation and the formation of tumors in the mice. Using several different strategies, we have compared the aromatase inhibitors (letrozole and anastrozole) with antiestrogens (tamoxifen and fulvestrant). Aromatase inhibitors were more effective than tamoxifen and were more effective alone than when combined with antiestrogens. In addition, letrozole had a longer duration of effect than tamoxifen. The possibility of delaying the development of resistance to antiestrogens and aromatase inhibitors was investigated by alternating letrozole with tamoxifen every 4 weeks. However, although alternating treatment proved more effective than tamoxifen alone (tumors doubled in 16 weeks), tumors doubled in volume at about 18 weeks when treatment began with tamoxifen. When treatment started with letrozole and alternated with tamoxifen, tumors doubled in volume in about 22 weeks. However, tumors of mice treated with letrozole alone did not double in size until 35 weeks. These results demonstrate that this aromatase inhibitor is more effective and has a longer duration of response as a single agent than tamoxifen or in combination with tamoxifen. PMID- 12538499 TI - Breast cancer endocrine resistance: how growth factor signaling and estrogen receptor coregulators modulate response. AB - Endocrine therapy, and especially tamoxifen, is the most important systemic treatment of estrogen receptor (ER)-positive breast cancer at all stages. A serious obstacle, however, is intrinsic or acquired resistance to these therapies, which in the case of selective ER modulators, such as tamoxifen, involves some imbalance of their agonist versus antagonist actions. Recent data suggest that levels of both ER coregulatory proteins and extra and intracellular signaling from growth factor-related pathways may be important in adjusting this mixed agonist/antagonist activity of selective ER modulators in resistant breast tumors. We suggest that ER coregulators' mediation of growth factor and other cellular signaling to the ER pathway is an important feature in endocrine response and resistance in breast cancer. Indeed, we find that failure of the antitumor activity of tamoxifen in patients with breast cancer is actually determined by both the levels of and the interaction between the ER coactivator amplified in breast cancer-1 (AIB1) and the epidermal growth factor-related protein HER. Thus, the interactions of these diverse elements are essential considerations in defining new predictive and therapeutic tools. PMID- 12538501 TI - Endocrine therapy of advanced disease: analysis and implications of the existing data. AB - Endocrine therapy remains an important approach to the treatment of metastatic breast cancer because of its effectiveness and excellent tolerability. In the last 10 years, a number of new endocrine therapies have been introduced. These include the luteinizing hormone-releasing hormone agonists, which produce menopausal changes in premenopausal women; the aromatase inhibitors, which prevent production of estrogen in postmenopausal women; and the estrogen receptor down-regulator fulvestrant (Faslodex), which is effective in postmenopausal women whose tumors have progressed following response to other selective estrogen receptor modulators. The endocrine cascade for the treatment of premenopausal women with metastatic disease now involves the concurrent or sequential combination of a luteinizing hormone-releasing hormone analogue and tamoxifen, whereas the cascade for the treatment of postmenopausal women can begin with tamoxifen followed by an aromatase inhibitor or with an aromatase inhibitor followed by tamoxifen. The optimal cascade following the use of an aromatase inhibitor and tamoxifen in postmenopausal women remains unclear, but fulvestrant and megestrol acetate or the use of an aromatase inactivator (exemestane) following an aromatase inhibitor are all available options with some activity. Over the next few years, clinical trials will clarify the optimal sequence of endocrine therapy for postmenopausal women. The use of estrogen and progesterone receptor status to select for endocrine therapy is undeniably crucial. HER2/neu overexpression may also predict response to endocrine therapy, but this remains controversial. PMID- 12538502 TI - Pharmacology and pharmacokinetics of the newer generation aromatase inhibitors. AB - The newer generation aromatase inhibitors (AIs) as a class show efficacy and tolerability benefits over previously established treatments in postmenopausal women with advanced breast cancer. At clinically administered doses, the plasma half-lives of anastrozole (1 mg once daily), letrozole (2.5 mg once daily), and exemestane (25 mg once daily) are 41-48 h, 2-4 days, and 27 h, respectively. Time to steady-state plasma levels is 7 days for both anastrozole and exemestane and 60 days for letrozole. Androgenic side effects have only been reported with exemestane. Anastrozole treatment has no impact on plasma lipid levels, whereas both letrozole and exemestane have an unfavorable effect. From indirect comparisons, anastrozole shows the highest degree of selectivity compared with letrozole and exemestane, in terms of a lack of effect on adrenosteroidogenesis. To date, there are no data suggesting any major differences in clinical efficacy between the newer generation AIs anastrozole and letrozole. Based on the observed pharmacological profiles, however, it cannot be assumed that the AIs will display the same tolerability and safety profiles when given for extended periods of time in the adjuvant setting. The effects of anastrozole, letrozole, and exemestane are being investigated in the adjuvant setting, and these data will elucidate the possible long-term consequences of the pharmacological effects reported after short-term exposure. PMID- 12538503 TI - Clinical differences among the aromatase inhibitors. AB - In the United States, three third-generation aromatase inhibitors are available commercially: anastrozole, letrozole, and exemestane. Anastrozole and letrozole are nonsteroidal agents, whereas exemestane is a steroid. The three agents differ in terms of structure and metabolic products and in the degree to which they suppress aromatase activity. The clinical significance of these differences is unclear. All three of the agents have been found to be equivalent or superior to megesterol acetate as a second-line therapy for metastatic breast cancer. In the first-line setting, large Phase III trials have demonstrated that anastrozole and letrozole are equivalent or superior to tamoxifen in women with metastatic disease. Multiple trials with widely varying study designs have been launched in the adjuvant setting comparing the aromatase inhibitors to tamoxifen. Early results from the ATAC (Arimidex, Tamoxifen, Alone or in Combination) trial suggest a small but statistically significant improvement in disease-free survival for anastrozole compared with tamoxifen, but further follow-up is needed. This article explores the efficacy and tolerability of the aromatase inhibitors in both the metastatic and the adjuvant settings. PMID- 12538504 TI - Adjuvant endocrine therapy in postmenopausal breast cancer. AB - Adjuvant endocrine therapy with tamoxifen has a clearly established benefit in postmenopausal women with resected early breast cancer that expresses the estrogen receptor and/or progesterone receptor. Whereas there is a vast and long experience with tamoxifen, the major focus of clinical trials over the past 6 years has involved the study of the third-generation aromatase inhibitors. Recently published data from the Arimidex, Tamoxifen, Alone or in Combination (ATAC) trial, which involved only postmenopausal women and is the largest adjuvant trial ever conducted, has demonstrated superior efficacy for anastrozole over tamoxifen alone or in combination with anastrozole. These data have engendered a great deal of discussion as to whether they provide a sufficient basis for changing the standard of practice in terms of choice of agent. Currently, a case can be made that further maturation of the ATAC trial is necessary and that tamoxifen should remain the standard of endocrine adjuvant therapy for most women, with anastrozole being used in those with a contraindication or intolerance to tamoxifen. However, close attention must be paid to new information as it develops not only related to efficacy but also to other end organ effects. PMID- 12538505 TI - Adjuvant hormonal therapy for premenopausal women with breast cancer. AB - Hormonal manipulation has been used for over 100 years to treat breast cancer. Ovarian ablation/suppression and tamoxifen are currently accepted adjuvant endocrine therapies for premenopausal breast cancer. Methods of permanently ablating ovarian function include surgical oophorectomy and radiation-induced ovarian failure; medical castration with luteinizing hormone-releasing hormone analogues is a reversible approach. Adjuvant chemotherapy frequently results in permanent amenorrhea and thus represents an indirect form of ovarian ablation. Although early randomized trials of ovarian ablation suffered from small sample sizes and design flaws, a meta-analysis of their results through the Early Breast Cancer Trialists' Collaborative Group demonstrated a clear benefit from ovarian ablation as a single intervention in the adjuvant treatment of women less than 50 years of age with breast cancer. The Early Breast Cancer Trialists' Collaborative Group meta-analysis also demonstrated the efficacy of 5 years of adjuvant tamoxifen as a single treatment modality, regardless of age. Even with these improvements in disease-free and overall survival, several important questions remain unanswered. The relative efficacy of adjuvant chemotherapy versus ovarian ablation/suppression has not been strictly defined. However, the data suggest that the clinical benefit of either chemotherapy or ovarian ablation/suppression and 5 years of tamoxifen is similar. Thus, ovarian ablation/suppression combined with tamoxifen is a reasonable alternative to chemotherapy for some women with good-risk early-stage breast cancer (high hormone receptor expression, low-grade or lymph node-negative disease), particularly those wishing to preserve fertility. The value of combining ovarian ablation/suppression with chemotherapy, other endocrine therapy, or both and ameliorating the long-term morbidity of estrogen deprivation remain important areas for investigation. With the advent of multiple targeted endocrine therapies with distinct mechanisms of action, there is a unique opportunity to design highly informative clinical trials that can define the optimal combinations and sequencing of hormonal therapies in the treatment of early-stage breast cancer. PMID- 12538506 TI - National surgical adjuvant breast and bowel project update: prevention trials and endocrine therapy of ductal carcinoma in situ. AB - Following up on the results of recent completed trials, several major breast cancer prevention trials are either underway or impending. In the Study of Tamoxifen and Raloxifene trial, eligible women are at least 35 years of age and postmenopausal, with either lobular carcinoma in situ or a 5-year risk of invasive breast cancer of at least 1.67%. The study will compare the ability of 5 years of tamoxifen or raloxifene to reduce the incidence of breast cancer. Subjects are randomly assigned to receive either 20 mg of tamoxifen or 60 mg of raloxifene daily. After 3 years of recruitment, 13647 women have been randomized (20.7% of those eligible). The median age of randomized women is 58 years (mean age, 58 years), and their median 5-year risk of breast cancer is 3.3% (mean 5 year risk of breast cancer, 4.0%). Hysterectomy was reported by 52.5% of the randomized women; lobular carcinoma in situ was reported by 8.4% of subjects before randomization. In the National Surgical Adjuvant Breast and Bowel Project (NSABP) B-24 trial, 1804 women with ductal carcinoma in situ were randomly assigned tamoxifen after lumpectomy and radiation therapy. Women in the tamoxifen group had fewer breast cancer events at 5 years than did those on placebo (8.2% versus 13.4%, P = 0.0009). The proposed NSABP B-35 trial will have the same design as NSABP B-24 but will compare tamoxifen with anastrozole in postmenopausal women. Outcomes will include both ipsilateral and contralateral new breast cancer and recurrences, as well as the occurrence of regional and distant disease. Enrollment will begin in early 2003. PMID- 12538507 TI - Preoperative models to evaluate endocrine strategies for breast cancer. AB - There are essentially two approaches to presurgical therapy. The first strategy is one in which the therapy is given to downstage the disease over a period of some 3-4 months. In these circumstances, biological studies can be conducted that can be associated with clinical response. The second strategy involves the institution of medical therapy before surgery with no delay to that surgery. This is essentially incidental and is not given with the aim of having a therapeutic benefit but rather for the biological study of the particular therapeutic approach. In the incidental therapy scenario, we have conducted a number of studies to evaluate the biological effects of raloxifene, idoxifene, fulvestrant (in comparison with tamoxifen), and the aromatase inhibitor 4 hydroxyandrostenedione. Significant reductions in proliferation were noted in all, as was down-regulation of estrogen receptor levels. The changes were most profound in those estrogen receptor-positive tumors that were also progesterone receptor positive, consistent with the greater clinical effect of these therapies in this population. This setting is particularly valuable for treatments in which there is no particular evidence for clinical benefit but in which the therapy is known to be safe. It is possible to evaluate potential resistance mechanisms by associating changes in Ki67 or apoptosis with the expression of the putative determinant of resistance in the short-term presurgical model. PMID- 12538508 TI - ErbB (HER) receptors can abrogate antiestrogen action in human breast cancer by multiple signaling mechanisms. AB - It has been reported that overexpression of the epidermal growth factor receptor (erbB1) or its homologous receptor, HER2 (erbB2), can confer antiestrogen resistance to estrogen receptor (ER)-positive human breast cancer cells. Aberrant signaling by receptors of the erbB network up-regulates a number of signaling pathways, which include phospholipase C-gamma1, Ras-Raf-mitogen-activated protein/extracellular signal-regulated kinase kinase-mitogen-activated protein kinase, phosphatidylinositol 3'-kinase and its target, the serine/threonine kinase Akt, stress-activated protein kinases, signal transducers and activators of transcription, and c-Jun-NH(2)-terminal kinase (JNK). Akt has been reported to induce estrogen-independent transcription of ER. Here we show that transfection of ER-positive, HER2 gene-amplified BT-74 cells with an expression vector encoding dominant-negative (K179M) Akt1 partially restored the ability of tamoxifen to inhibit estradiol-stimulated ER reporter activity. Infection of MCF 7 cells with an adenovirus encoding myristoylated, constitutively active Akt induced ER reporter activity in the absence of estradiol and resulted in tamoxifen resistance of these cells in culture. Data will be presented to suggest that, in addition to mitogen-activated protein kinase, Akt is an important mediator of HER2-mediated antiestrogen resistance in human breast cancer cells. PMID- 12538509 TI - New mechanisms of signal transduction inhibitor action: receptor tyrosine kinase down-regulation and blockade of signal transactivation. AB - The explosion of signal transduction research over the last 10 years has provided a unique insight into the complexity of these intricate pathways. Whereas intermediates of multiple signaling pathways have been identified, understanding their function and, in particular, the interactions between them has become a daunting task. The increasing evidence that many of these pathways can cross-talk with each other via signal transactivation inevitably raises the question of how cells determine specificity in signaling. Despite the mind-numbing complexity of these pathways, progress has been made in developing highly specific and potent signal transduction inhibitors (STIs). STIs show promise in the treatment of cancer in preclinical studies and are currently in a number of clinical trials. Whereas many of these agents were "rationally designed," we barely understand their mechanisms of action. This review will highlight how recent studies using these STIs have elucidated novel mechanisms of STI action that may be used in the development of new therapeutic strategies for the treatment of cancer. PMID- 12538510 TI - Integration of signal transduction inhibitors with endocrine therapy: an approach to overcoming hormone resistance in breast cancer. AB - Recent evidence suggests that common molecular adaptations occur during resistance to both tamoxifen and estrogen deprivation that use various signal transduction pathways, often involving cross-talk with a retained and functional estrogen receptor (ER) protein. There appear to be several different levels at which this cross-talk may occur, including peptide growth factor signaling via the type 1 tyrosine kinase growth factor receptor family [epidermal growth factor receptor (EGFR) and HER2], which may become up-regulated during endocrine treatment, ultimately being harnessed by cells to allow them hormone-independent growth. ER may remain involved in cell growth with ligand-independent phosphorylation and activation via different intracellular mitogen-activated protein kinases. ER may also become involved in non-nuclear estrogen-dependent signaling via interaction with the phosphatidylinositol 3'-kinase/Akt cell survival pathway or may interact with the stress-activated protein kinase/c-Jun NH(2)-terminal kinase pathway. Understanding these mechanisms will permit the optimal integration of new signal transduction inhibitors (STIs) into breast cancer therapy. Preclinical approaches that have shown promise include the use of EGFR tyrosine kinase inhibitors for hormone-resistant breast cancer cells that are dependent on either EGFR or HER2 signaling. Likewise, farnesyl transferase inhibitors, mitogen-activated protein kinase inhibitors, and cell cycle inhibitors have all shown activity in experimental breast cancer models. Emerging data suggest that STIs may be more effective when given in combination with endocrine therapy either to overcome resistance or to prevent/delay emergence of the resistance phenotype. Clinical trials are in progress to determine the safety and optimal schedule for each of the various STIs, and studies of STIs in combination with aromatase inhibitors have commenced in breast cancer to see whether the therapeutic response to endocrine therapy can be enhanced further. PMID- 12538511 TI - Temporal and cellular requirements for Fms signaling during zebrafish adult pigment pattern development. AB - Ectothermic vertebrates exhibit a diverse array of adult pigment patterns. A common element of these patterns is alternating dark and light stripes each comprising different classes of neural crest-derived pigment cells. In the zebrafish, Danio rerio, alternating horizontal stripes of black melanophores and yellow xanthophores are a prominent feature of the adult pigment pattern. In fms mutant zebrafish, however, xanthophores fail to develop and melanophore stripes are severely disrupted. fms encodes a type III receptor tyrosine kinase expressed by xanthophores and their precursors and is the closest known homologue of kit, which has long been studied for roles in pigment pattern development in amniotes. In this study we assess the cellular and temporal requirements for Fms activity in promoting adult pigment pattern development. By transplanting cells between fms mutants and either wild-type or nacre mutant zebrafish, we show that fms acts autonomously to the xanthophore lineage in promoting the striped arrangement of adult melanophores. To identify critical periods for fms activity, we isolated temperature sensitive alleles of fms and performed reciprocal temperature shift experiments at a range of stages from embryo to adult. These analyses demonstrate that Fms is essential for maintaining cells of the xanthophore lineage as well as maintaining the organization of melanophore stripes throughout development. Finally, we show that restoring Fms activity even at late larval stages allows essentially complete recovery of xanthophores and the development of a normal melanophore stripe pattern. Our findings suggest that fms is not required for establishing a population of precursor cells during embryogenesis but is required for recruiting pigment cell precursors to xanthophore fates, with concomitant effects on melanophore organization. PMID- 12538512 TI - Orb and a long poly(A) tail are required for efficient oskar translation at the posterior pole of the Drosophila oocyte. AB - During Drosophila oogenesis, the posterior determinant, Oskar, is tightly localized at the posterior pole of the oocyte. The exclusive accumulation of Oskar at this site is ensured by localization-dependent translation of oskar mRNA: translation of oskar mRNA is repressed during transport and activated upon localization at the posterior cortex. Previous studies have suggested that oskar translation is poly(A)-independent. We show that a long poly(A) tail is required for efficient oskar translation, both in vivo and in vitro, but is not sufficient to overcome BRE-mediated repression. Moreover, we show that accumulation of Oskar activity requires the Drosophila homolog of Cytoplasmic Polyadenylation Element Binding protein (CPEB), Orb. As posterior localization of oskar mRNA is an essential prerequisite for its translation, it was critical to identify an allele of orb that does localize oskar mRNA to the posterior pole of the oocyte. We show that flies bearing the weak mutation orb(mel) localize oskar transcripts with a shortened poly(A) that fails to enhance oskar translation, resulting in reduced Oskar levels and posterior patterning defects. We conclude that Orb-mediated cytoplasmic polyadenylation stimulates oskar translation to achieve the high levels of Oskar protein necessary for posterior patterning and germline differentiation. PMID- 12538513 TI - CRM1-mediated nuclear export and regulated activity of the Receptor Tyrosine Kinase antagonist YAN require specific interactions with MAE. AB - ETS family transcription factors serve as downstream effectors of signal transduction pathways, mediating cellular proliferation, differentiation and, when misregulated, tumorigenesis. The transcriptional repressor YAN prevents inappropriate responses to Receptor Tyrosine Kinase signaling by outcompeting POINTED for access to target gene promoters. We demonstrate that the molecular mechanism underlying downregulation of YAN involves CRM1-mediated nuclear export and define a novel role in this context for MAE, a co-factor previously implicated in facilitating MAPK phosphorylation of YAN. In addition to promoting YAN downregulation, MAE also participates in an inhibitory feedback loop that attenuates POINTED-P2 activation. Thus, we propose that MAE plays multiple independent roles in fine-tuning the levels of POINTED and YAN activity in accordance with changing RTK signaling conditions. PMID- 12538514 TI - Maelstrom, a Drosophila spindle-class gene, encodes a protein that colocalizes with Vasa and RDE1/AGO1 homolog, Aubergine, in nuage. AB - A hallmark of germline cells across the animal kingdom is the presence of perinuclear, electron-dense granules called nuage. In many species examined, Vasa, a DEAD-box RNA helicase, is found in these morphologically distinct particles. Despite its evolutionary conservation, the function of nuage remains obscure. We have characterized a null allele of maelstrom (mael) and shown that Maelstrom protein is localized to nuage in a Vasa-dependent manner. By phenotypic characterization, we have defined maelstrom as a spindle-class gene that affects Vasa modification. In a nuclear transport assay, we have determined that Maelstrom shuttles between the nucleus and cytoplasm, which may indicate a nuclear origin for nuage components. Interestingly, Maelstrom, but not Vasa, depends on two genes involved in RNAi phenomena, aubergine and spindle-E (spn-E), for its nuage localization. Furthermore, maelstrom mutant ovaries show mislocalization of two proteins involved in the microRNA and/or RNAi pathways, Dicer and Argonaute2, suggesting a potential connection between nuage and the microRNA-pathway. PMID- 12538515 TI - Shaping the zebrafish notochord. AB - Promptly after the notochord domain is specified in the vertebrate dorsal mesoderm, it undergoes dramatic morphogenesis. Beginning during gastrulation, convergence and extension movements change a squat cellular array into a narrow, elongated one that defines the primary axis of the embryo. Convergence and extension might be coupled by a highly organized cellular intermixing known as mediolateral intercalation behavior (MIB). To learn whether MIB drives early morphogenesis of the zebrafish notochord, we made 4D recordings and quantitatively analyzed both local cellular interactions and global changes in the shape of the dorsal mesodermal field. We show that MIB appears to mediate convergence and can account for extension throughout the dorsal mesoderm. Comparing the notochord and adjacent somitic mesoderm reveals that extension can be regulated separately from convergence. Moreover, mutational analysis shows that extension does not require convergence. Hence, a cellular machine separate from MIB that can drive dorsal mesodermal extension exists in the zebrafish gastrula. The likely redundant control of morphogenesis may provide for plasticity at this critical stage of early development. PMID- 12538516 TI - Composition and dynamics of the Caenorhabditis elegans early embryonic transcriptome. AB - Temporal profiles of transcript abundance during embryonic development were obtained by whole-genome expression analysis from precisely staged C. elegans embryos. The result is a highly resolved time course that commences with the zygote and extends into mid-gastrulation, spanning the transition from maternal to embryonic control of development and including the presumptive specification of most major cell fates. Transcripts for nearly half (8890) of the predicted open reading frames are detected and expression levels for the majority of them (>70%) change over time. The transcriptome is stable up to the four-cell stage where it begins rapidly changing until the rate of change plateaus before gastrulation. At gastrulation temporal patterns of maternal degradation and embryonic expression intersect indicating a mid-blastula transition from maternal to embryonic control of development. In addition, we find that embryonic genes tend to be expressed transiently on a time scale consistent with developmental decisions being made with each cell cycle. Furthermore, overall rates of synthesis and degradation are matched such that the transcriptome maintains a steady-state frequency distribution. Finally, a versatile analytical platform based on cluster analysis and developmental classification of genes is provided. PMID- 12538517 TI - Orbit/Mast, the CLASP orthologue of Drosophila, is required for asymmetric stem cell and cystocyte divisions and development of the polarised microtubule network that interconnects oocyte and nurse cells during oogenesis. AB - Drosophila oocyte differentiation is preceded by the formation of a polarised 16 cell cyst from a single progenitor stem cell as a result of four rounds of asymmetric mitosis followed by incomplete cytokinesis. We show that the Orbit/Mast microtubule-associated protein is required at several stages in the formation of such polarised 16-cell cysts. In wild-type cysts, the Orbit/Mast protein not only associates with the mitotic spindle and its poles, but also with the central spindle (spindle remnant), ring canal and fusome, suggesting it participates in interactions between these structures. In orbit mutants, the stem cells and their associated fusomes are eventually lost as Orbit/Mast protein is depleted. The mitotic spindles of those cystocytes that do divide are either diminutive or monopolar, and do not make contact with the fusome. Moreover, the spindle remnants and ring canals fail to differentiate correctly in such cells and the structure of fusome is compromised. The Orbit/Mast protein thus appears to facilitate multiple interactions of the fusome with mitotic spindles and ring canals. This ensures correct growth of the fusome into a branched asymmetrically distributed organelle that is pre-determinative of 16-cell cyst formation and oocyte fate specification. Finally the Orbit/Mast protein is required during mid oogenesis for the organisation of the polarised microtubule network inside the 16 cell cyst that ensures oocyte differentiation. The localisation of CLIP-190 to such microtubules and to the fusome is dependent upon Orbit/Mast to which it is complexed. PMID- 12538518 TI - Differential function of RNCAM isoforms in precise target selection of olfactory sensory neurons. AB - Olfactory sensory neurons (OSNs) are individually specified to express one odorant receptor (OR) gene among approximately 1000 different and project with precision to topographically defined convergence sites, the glomeruli, in the olfactory bulb. Although ORs partially determine the location of convergence sites, the mechanism ensuring that axons with different OR identities do not co converge is unknown. RNCAM (OCAM, NCAM2) is assumed to regulate a broad zonal segregation of projections by virtue of being a homophilic cell adhesion molecule that is selectively expressed on axons terminating in a defined olfactory bulb region. We have identified NADPH diaphorase activity as being an independent marker for RNCAM-negative axons. Analyses of transgenic mice that ectopically express RNCAM in NADPH diaphorase-positive OSNs show that the postulated function of RNCAM in mediating zone-specific segregation of axons is unlikely. Instead, analyses of one OR-specific OSN subpopulation (P2) reveal that elevated RNCAM levels result in an increased number of P2 axons that incorrectly co-converge with axons of other OR identities. Both Gpi-anchored and transmembrane-bound RNCAM isoforms are localized on axons in the nerve layer, while the transmembrane bound RNCAM is the predominant isoform on axon terminals within glomeruli. Overexpressing transmembrane-bound RNCAM results in co-convergence events close to the correct target glomeruli. By contrast, overexpression of Gpi-anchored RNCAM results in axons that can bypass the correct target before co-converging on glomeruli located at a distance. The phenotype specific for Gpi-anchored RNCAM is suppressed in mice overexpressing both isoforms, which suggests that two distinct RNCAM isoform-dependent activities influence segregation of OR-defined axon subclasses. PMID- 12538519 TI - Zebrafish foxi1 mediates otic placode formation and jaw development. AB - The otic placode is a transient embryonic structure that gives rise to the inner ear. Although inductive signals for otic placode formation have been characterized, less is known about the molecules that respond to these signals within otic primordia. Here, we identify a mutation in zebrafish, hearsay, which disrupts the initiation of placode formation. We show that hearsay disrupts foxi1, a forkhead domain-containing gene, which is expressed in otic precursor cells before placodes become visible; foxi1 appears to be the earliest marker known for the otic anlage. We provide evidence that foxi1 regulates expression of pax8, indicating a very early role for this gene in placode formation. In addition, foxi1 is expressed in the developing branchial arches, and jaw formation is disrupted in hearsay mutant embryos. PMID- 12538520 TI - Neural crest patterning: autoregulatory and crest-specific elements co-operate for Krox20 transcriptional control. AB - Neural crest patterning constitutes an important element in the control of the morphogenesis of craniofacial structures. Krox20, a transcription factor gene that plays a critical role in the development of the segmented hindbrain, is expressed in rhombomeres (r) 3 and 5 and in a stream of neural crest cells migrating from r5 toward the third branchial arch. We have investigated the basis of the specific neural crest expression of Krox20 and identified a cis-acting enhancer element (NCE) located 26 kb upstream of the gene that is conserved between mouse, man and chick and can recapitulate the Krox20 neural crest pattern in transgenic mice. Functional dissection of the enhancer revealed the presence of two conserved Krox20 binding sites mediating direct Krox20 autoregulation in the neural crest. In addition, the enhancer included another essential element containing conserved binding sites for high mobility group (HMG) box proteins and which responded to factors expressed throughout the neural crest. Consistent with this the NCE was strongly activated in vitro by Sox10, a crest-specific HMG box protein, in synergism with Krox20, and the inactivation of Sox10 prevented the maintenance of Krox20 expression in the migrating neural crest. These results suggest that the dependency of the enhancer on both crest- (Sox10) and r5- (Krox20) specific factors limits its activity to the r5-derived neural crest. This organisation also suggests a mechanism for the transfer and maintenance of rhombomere-specific gene expression from the hindbrain neuroepithelium to the emerging neural crest and may be of more general significance for neural crest patterning. PMID- 12538521 TI - Hedgehog signalling maintains the optic stalk-retinal interface through the regulation of Vax gene activity. AB - During early formation of the eye, the optic vesicle becomes partitioned into a proximal domain that forms the optic nerve and a distal domain that forms the retina. In this study, we investigate the activity of Nodal, Hedgehog (Hh) and Fgf signals and Vax family homeodomain proteins in this patterning event. We show that zebrafish vax1 and vax2 are expressed in overlapping domains encompassing the ventral retina, optic stalks and preoptic area. Abrogation of Vax1 and Vax2 activity leads to a failure to close the choroid fissure and progressive expansion of retinal tissue into the optic nerve, finally resulting in a fusion of retinal neurons and pigment epithelium with forebrain tissue. We show that Hh signals acting through Smoothened act downstream of the Nodal pathway to promote Vax gene expression. However, in the absence of both Nodal and Hh signals, Vax genes are expressed revealing that other signals, which we show include Fgfs, contribute to Vax gene regulation. Finally, we show that Pax2.1 and Vax1/Vax2 are likely to act in parallel downstream of Hh activity and that the bel locus (yet to be cloned) mediates the ability of Hh-, and perhaps Fgf-, signals to induce Vax expression in the preoptic area. Taking all these results together, we present a model of the partitioning of the optic vesicle along its proximo-distal axis. PMID- 12538522 TI - Analysis of the mutational effects of the COP/DET/FUS loci on genome expression profiles reveals their overlapping yet not identical roles in regulating Arabidopsis seedling development. AB - Microarray gene expression profiling was used to examine the role of pleiotropic COP/DET/FUS loci as well as other partially photomorphogenic loci during Arabidopsis seedling development and genome expression regulation. Four types of lethal, pleiotropic cop/det/fus mutants exhibit qualitatively similar gene expression profiles, yet each has specific differences. Mutations in COP1 and DET1 show the most similar genome expression profiles, while the mutations in the COP9 signalosome (CSN) and COP10 exhibit increasingly diverged genome expression profiles in both darkness and light. The genome expression profiles of the viable mutants of COP1 and DET1 in darkness mimic those of the physiological light regulated genome expression profiles, whereas the genome expression profiles of representative lethal mutants belong to another clade and significantly diverge from the normal light control of genome expression. Instead, these lethal pleiotropic mutants show genome expression profiles similar to those from seedlings growth under high light intensity stress. Distinct lethal pleiotropic cop/det/fus mutants also result in distinct expression profiles in the small portion of genes examined and exhibit similar relatedness in both light and darkness. The partial cop/det/fus mutants affected expression of both light regulated and non-light regulated genes. Our results suggest that pleiotropic COP/DET/FUS loci control is largely overlapping but also has separable roles in plant development. The partially photomorphogenic loci regulate a subset of photomorphogenic responses as well as other non-light regulated processes. PMID- 12538523 TI - Functional specificity of the nematode Hox gene mab-5. AB - Hox genes encode evolutionarily conserved transcription factors involved in morphological specification along the anteroposterior body axis of animals. The two most striking features of Hox genes are colinearity and the strong sequence conservation. Among all animals studied so far, the nematode Caenorhabditis elegans contains one of the most divergent Hox clusters. The core cluster contains only four members, which in part deviate from the colinearity rule. In addition, orthologous and paralogous nematode Hox sequences diverged substantially. Given these nematode-specific features, we asked how these Hox proteins evolved and how they provide functional specificity. We investigated the role of MAB-5 during ray formation and established an in vivo assay using Cel-mab 5 regulatory elements to express orthologous, paralogous and chimeric cDNAs in a Cel-mab-5 mutant background. We show that the MAB-5 ortholog from Pristionchus pacificus, but not the C. elegans paralogous Hox proteins can rescue Cel-mab-5. Experiments with chimeric, truncated and mutagenized Hox proteins suggest the specificity to be conferred by the N-terminal arm and helix I, but not helix II of the homeodomain. PMID- 12538524 TI - Macrophage stimulating protein is a target-derived neurotrophic factor for developing sensory and sympathetic neurons. AB - Macrophage stimulating protein (MSP) is a pleiotropic growth factor that signals via the Ron receptor tyrosine kinase. We report that Ron mRNA is expressed by NGF dependent sensory and sympathetic neurons and that these neurons survive and grow with MSP at different stages of development. Whereas NGF-dependent sensory neurons become increasingly responsive to MSP with age, sympathetic neurons exhibit an early response to MSP that is lost by birth. MSP mRNA expression increases with age in sensory neuron targets and decreases in sympathetic targets. After the phase of naturally occurring neuronal death, significant numbers of NGF-dependent sensory neurons, but not sensory neurons, dependent on other neurotrophins, are lost in mice lacking a functional Ron receptor. These results show that MSP is a target-derived neurotrophic factor for subsets of sensory and sympathetic neurons at different times during their development. PMID- 12538525 TI - Wnt5a and Wnt5b exhibit distinct activities in coordinating chondrocyte proliferation and differentiation. AB - Proper longitudinal growth of long bones relies on the regulation of specific spatial patterns of chondrocyte proliferation and differentiation. We have studied the roles of two members of the Wnt family, Wnt5a and Wnt5b in long bone development. We show that Wnt5a is required for longitudinal skeletal outgrowth and that both Wnt5a and Wnt5b regulate the transition between different chondrocyte zones independently of the Indian hedgehog (Ihh)/parathyroid hormone related peptide (PTHrP) negative feedback loop. We find that important cell cycle regulators such as cyclin D1 and p130, a member of the retinoblastoma family, exhibit complimentary expression patterns that correlate with the distinct proliferation and differentiation states of chondrocyte zones. Furthermore, we show that Wnt5a and Wnt5b appear to coordinate chondrocyte proliferation and differentiation by differentially regulating cyclin D1 and p130 expression, as well as chondrocyte-specific Col2a1 expression. Our data indicate that Wnt5a and Wnt5b control the pace of transitions between different chondrocyte zones. PMID- 12538526 TI - Apoptosis-mediated cell death within the ovarian polar cell lineage of Drosophila melanogaster. AB - Polar cells have been described as pairs of specific follicular cells present at each pole of Drosophila egg chambers. They are required at different stages of oogenesis for egg chamber formation and establishment of both the anteroposterior and planar polarities of the follicular epithelium. We show that definition of polar cell pairs is a progressive process since early stage egg chambers contain a cluster of several polar cell marker-expressing cells at each pole, while as of stage 5, they contain invariantly two pairs of such cells. Using cell lineage analysis, we demonstrate that these pre-polar cell clusters have a polyclonal origin and derive specifically from the polar cell lineage, rather than from that giving rise to follicular cells. In addition, selection of two polar cells from groups of pre-polar cells occurs via an apoptosis-dependent mechanism and is required for correct patterning of the anterior follicular epithelium of vitellogenic egg chambers. PMID- 12538527 TI - Dax1 regulates testis cord organization during gonadal differentiation. AB - Mutations of the DAX1 nuclear receptor gene cause adrenal hypoplasia congenita, an X-linked disorder characterized by adrenal insufficiency and hypogonadotropic hypogonadism. Targeted deletion of Dax1 in mice also reveals primary testicular dysgenesis, which is manifest by obstruction of the rete testis by Sertoli cells and hyperplastic Leydig cells, leading to seminiferous tubule dilation and degeneration of germ cells. Because Dax1 is expressed early in gonadal development, and because Sertoli and Leydig cells are located ectopically in the adult, we hypothesized that these testis abnormalities are the result of an early defect in testis development. In Dax1(-/Y) males, the gonad develops normally until 12.5 dpc. However, by 13.5 dpc, the testis cords are disorganized and incompletely formed in Dax1-deficient mice. The number of germ and Sertoli cells is unchanged, and the expression of Sertoli-specific markers appears to be normal. However, the number of peritubular myoid cells, which normally surround the testis cords, is reduced. BrdU labeling of peritubular myoid cells is low, consistent with decreased proliferation. The basal lamina produced by peritubular myoid and Sertoli cells is disrupted, leading to open and incompletely formed testis cords. Leydig cells, which normally reside in the peritubular space and extend from the coelomic surface to the dorsal surface of the gonad, are restricted to the coelomic surface of Dax1-deficient testis. We conclude that Dax1 plays a crucial role in testis differentiation by regulating the development of peritubular myoid cells and the formation of intact testis cords. The developmental abnormalities in the Dax1-deficient testis lay the foundation for gonadal dysgenesis and infertility in adult mice and, potentially in humans with DAX1 mutations. PMID- 12538528 TI - The primacy of primary care: reading Romanow. PMID- 12538531 TI - CANM endorsement of densitometry guidelines. PMID- 12538532 TI - Protection after medical error. PMID- 12538533 TI - Drug marketing priorities. PMID- 12538534 TI - Neuroradiologists and stroke. PMID- 12538538 TI - Pharmacists helping physicians. PMID- 12538540 TI - Family health networks in Ontario. PMID- 12538541 TI - Setting the record straight. PMID- 12538542 TI - Shaken baby syndrome in Canada: clinical characteristics and outcomes of hospital cases. AB - BACKGROUND: Shaken baby syndrome is an extremely serious form of abusive head trauma, the extent of which is unknown in Canada. Our objective was to describe, from a national perspective, the clinical characteristics and outcome of children admitted to hospital with shaken baby syndrome. METHODS: We performed a retrospective chart review, for the years 1988-1998, of the cases of shaken baby syndrome that were reported to the child protection teams of 11 pediatric tertiary care hospitals in Canada. Shaken baby syndrome was defined as any case reported at each institution of intracranial, intraocular or cervical spine injury resulting from a substantiated or suspected shaking, with or without impact, in children aged less than 5 years. RESULTS: The median age of subjects was 4.6 months (range 7 days to 58 months), and 56% were boys. Presenting complaints for the 364 children identified as having shaken baby syndrome were nonspecific (seizure-like episode [45%], decreased level of consciousness [43%] and respiratory difficulty [34%]), though bruising was noted on examination in 46%. A history and/or clinical evidence of previous maltreatment was noted in 220 children (60%), and 80 families (22%) had had previous involvement with child welfare authorities. As a direct result of the shaking, 69 children died (19%) and, of those who survived, 162 (55%) had ongoing neurological injury and 192 (65%) had visual impairment. Only 65 (22%) of those who survived were considered to show no signs of health or developmental impairment at the time of discharge. INTERPRETATION: Shaken baby syndrome results in an extremely high degree of mortality and morbidity. Ongoing care of these children places a substantial burden on the medical system, caregivers and society. PMID- 12538543 TI - Screening for diabetic retinopathy in James Bay, Ontario: a cost-effectiveness analysis. AB - BACKGROUND: Retinopathy is a common complication of diabetes mellitus that if detected early by regular retinal examinations can be treated; thus, blindness can be delayed or prevented. Providing high-quality retinal screening is difficult, especially in rural and remote areas. Canada's First Nations population has a higher prevalence of diabetes and is, in general, more geographically isolated than the broader Canadian population. We modelled the cost-effectiveness of retinopathy screening by travelling retina specialists versus retinal photography with a portable digital camera in an isolated First Nations cohort with diabetes. METHODS: The 2 screening programs were modelled to run concurrently for 5 years, with outcomes evaluated over 10 years. To construct economic models for the population of Cree individuals living in the western James Bay area of northern Ontario, we used available data on the prevalence of diabetes in the area and estimates of the incidence of retinopathy derived from the published literature. We compared the screening models and calculated total costs, visual outcome, costs per sight-year saved and costs per quality-adjusted life year (QALY). We also estimated the costs of implementing a screening program for all First Nations individuals in Ontario with diabetes and no access to an ophthalmologist. RESULTS: From the perspective of the health care system the camera program was preferable to the specialist-based program. Over 10 years, 67 v. 56 sight years were saved, compared with no screening, at costs of 3900 Canadian dollars v. 9800 Canadian dollars per sight year and 15,000 Canadian dollars v. 37,000 Canadian dollars per QALY. Generalizing these results to the province of Ontario, the camera system could allow most isolated First Nations people with diabetes to be screened for 5 years for approximately 1.2 million Canadian dollars. INTERPRETATION: A portable retinal camera is a cost-effective means of screening for diabetic retinopathy in isolated communities of at-risk individuals. PMID- 12538544 TI - Impact of supply-side policies for control of illicit drugs in the face of the AIDS and overdose epidemics: investigation of a massive heroin seizure. AB - BACKGROUND: More than 93% of the nearly $500 million spent annually on Canada's drug strategy goes toward efforts to reduce the illicit drug supply. However, little is known about the effectiveness of this strategy. On Sept. 2, 2000, Canadian police seized approximately 100 kg of heroin in one of the nation's largest-ever seizures of this drug. An ongoing prospective cohort study of injection drug users afforded an opportunity to evaluate the impact of this seizure. METHODS: The Vancouver Injection Drug User Study is a prospective cohort study of injection drug users that began in 1996. The present study relied primarily on data acquired from participants who were seen during the 30-day periods immediately before and after the seizure. We compared drug use and behavioural characteristics, heroin and cocaine prices, and participants' reports of whether law enforcement had affected their source of drugs or the types of drugs available on the street, as well as overdoses, in these 2 periods. RESULTS: The 138 participants seen before the seizure were similar to the 123 participants seen after the seizure with respect to age, sex, ethnic background, education, HIV serostatus, neighbourhood residence, instability of housing, employment status, use of methadone maintenance therapy and all other measured potential confounders (all p > 0.10). We found no difference in the extent to which participants in the 2 groups reported daily use of heroin, frequency of nonfatal overdoses, or whether law enforcement had affected their source of drugs or the types of drugs available on the street (all p > 0.10). Although we detected no difference in the price of cocaine, the median reported price of heroin went down after the seizure (p = 0.034), which suggests that other shipments compensated for the seizure. External evaluations of deaths from overdoses and heroin purity indicated that the seizure had no impact, nor was any impact seen when the periods of analysis were extended. INTERPRETATION: The massive heroin seizure appeared to have no measurable public health benefit. Closer scrutiny of enforcement efforts is warranted to ensure that resources are delivered to the most efficient and cost-effective public health programs. PMID- 12538545 TI - Hypereosinophilic syndrome with pulmonary and cardiac involvement in a patient with asthma. AB - Idiopathic hypereosinophilic syndrome is characterized by prolonged eosinophilia without an identifiable underlying cause and multiple-organ dysfunction, most frequently involving the heart, the central or peripheral nervous system and the lungs. We describe a case in which a patient with asthma who had idiopathic hypereosinophilic syndrome with pulmonary involvement presented with symptoms of pulmonary embolism and left ventricular thrombus. PMID- 12538546 TI - Colorectal cancer screening in Canada: it's time to act. PMID- 12538547 TI - Complementary or alternative medicine: the need for plausibility. PMID- 12538550 TI - Active hand warming eases peripheral intravenous catheter insertion. PMID- 12538548 TI - Diagnosis of pulmonary embolism. AB - No single noninvasive test for pulmonary embolism is both sensitive and specific. Some tests are good for "ruling in" pulmonary embolism (e.g., helical CT) and some tests are good for "ruling out" pulmonary embolism (e.g., D-dimer); others are able to do both but are often nondiagnostic (e.g., ventilation-perfusion lung scanning). For optimal efficiency, choice of the initial diagnostic test should be guided by clinical assessment of the probability of pulmonary embolism and by patient characteristics that may influence test accuracy. This selective approach to testing enables pulmonary embolism to be diagnosed or excluded in a minimum number of steps. However, even with the appropriate use of combinations of noninvasive tests, it is often not possible to definitively diagnose or exclude pulmonary embolism at initial presentation. Most of these patients can be managed safely without treatment or pulmonary angiography by repeating ultrasound testing of the proximal veins after one and 2 weeks to detect evolving deep vein thrombosis. Helical CT and MRI are rapidly improving as diagnostic tests for pulmonary embolism and are expected to become central to its evaluation. PMID- 12538549 TI - Symptomatic hyperlactatemia in an HIV-positive patient: a case report and discussion. AB - Prolonged exposure to highly active antiretroviral therapy may be associated with adverse effects related to mitochondrial toxicity, such as hyperlactatemia. We describe a case of symptomatic hyperlactatemia in an HIV-positive patient to illustrate the subtle clinical symptoms and abnormal laboratory test results associated with this condition. We also review the pathophysiology, prevalence, spectrum and management of disturbances in lactate homeostasis induced by nucleoside reverse transcriptase inhibitors. PMID- 12538551 TI - Pink ladies: mercury poisoning in twin girls. PMID- 12538559 TI - Canada will check donor blood for West Nile virus if test available. PMID- 12538560 TI - Edmonton tackles shaken baby syndrome. PMID- 12538561 TI - US OKs new rapid HIV test, approval to be sought in Canada. PMID- 12538563 TI - Europe rejects pitch for direct-to-consumer drug ads. PMID- 12538565 TI - CIHR floats "research-into-action" trial balloon. PMID- 12538568 TI - Clinic beats Romanow to the primary care punch. PMID- 12538575 TI - Granulocyte-macrophage colony-stimulating factor signals for increased glucose transport via phosphatidylinositol 3-kinase- and hydrogen peroxide-dependent mechanisms. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) stimulates cellular glucose uptake by decreasing the apparent K(m) for substrate transport through facilitative glucose transporters on the plasma membrane. Little is known about this signal transduction pathway and the role of the alpha subunit of the GM-CSF receptor (alpha GMR) in modulating transporter activity. We examined the function of phosphatidylinositol 3-kinase (PI 3-kinase) in GM-CSF-stimulated glucose uptake and found that PI 3-kinase inhibitors, wortmannin and LY294002, completely blocked the GM-CSF-dependent increase of glucose uptake in Xenopus oocytes expressing the low affinity alpha GMR and in human cells expressing the high affinity alpha beta GMR complex. We identified a Src homology 3 domain-binding motif in alpha GMR at residues 358-361 as a potential interaction site for the PI 3-kinase regulatory subunit, p85. Physical evidence for p85 binding to alpha GMR was obtained by co-immunoprecipitation with antibodies to alpha GMR and p85, and an alpha GMR mutant with alteration of the Src homology 3 binding domain lost the ability to bind p85. Experiments with a construct eliminating most of the intracellular portion of alpha GMR showed a 50% reduction in GM-CSF-stimulated glucose uptake with residual activity blocked by wortmannin. Searching for a proximally generated diffusible factor capable of activating PI 3-kinase, we identified hydrogen peroxide (H(2)O(2)), generated by ligand or antibody binding to alpha GMR, as the initiating factor. Catalase treatment abrogated GM-CSF- or anti-alpha GMR antibody-stimulated glucose uptake in alpha GMR-expressing oocytes, and H(2)O(2) activated PI 3-kinase and led to some stimulation of glucose uptake in uninjected oocytes. Human myeloid cell lines and primary explant human lymphocytes expressing high affinity GM-CSF receptors responded to alpha GMR antibody with increased glucose uptake. These results identify the early events in the stimulation of glucose uptake by GM-CSF as involving local H(2)O(2) generation and requiring PI 3-kinase activation. Our findings also provide a mechanistic explanation for signaling through the isolated alpha subunit of the GM-CSF receptor. PMID- 12538576 TI - All six modules of the gelatin-binding domain of fibronectin are required for full affinity. AB - The gelatin-binding sites of fibronectin are confined to a 42-kDa region having four type I and two type II modules in the following order: I(6)-II(1)-II(2)-I(7) I(8)-I(9). To determine the relative importance of each module for recognition of gelatin, recombinant green fluorescent fusion proteins were prepared in which individual modules or groups of modules were deleted, and the resulting proteins were tested for binding to gelatin by analytical affinity chromatography. Deletion of both type II modules did not eliminate binding, confirming that at least some of the type I modules in this region are able to bind gelatin. It was found that deletion of type I module 6 tends to increase the affinity, whereas deletion of any other module decreases it. Deletion of module I(9) had a large effect but only if module II(2) was also present, suggesting an interaction between these two noncontiguous modules. Analysis of more than 20 recombinant fusion products led to the conclusion that all modules contribute to the interaction either directly by contacting the ligand or indirectly through module module interactions. PMID- 12538577 TI - Mapping interactions between the Ca2+-ATPase and its substrate ATP with infrared spectroscopy. AB - Infrared spectroscopy has been used to map substrate-protein interactions: the conformational changes of the sarcoplasmic reticulum Ca(2+)-ATPase upon nucleotide binding and ATPase phosphorylation were monitored using the substrate ATP and ATP analogues (2'-deoxy-ATP, 3'-deoxy-ATP, and inosine 5'-triphosphate), which were modified at specific functional groups of the substrate. Modifications to the 2'-OH, the 3'-OH, and the amino group of adenine reduce the extent of binding-induced conformational change of the ATPase, with particularly strong effects observed for the latter two. This demonstrates the structural sensitivity of the nucleotide-ATPase complex to individual interactions between nucleotide and ATPase. All groups studied are important for binding and interactions of a given ligand group with the ATPase depend on interactions of other ligand groups. Phosphorylation of the ATPase was observed for ITP and 2'-deoxy-ATP, but not for 3'-deoxy-ATP. There is no direct link between the extent of conformational change upon nucleotide binding and the rate of phosphorylation showing that the full extent of the ATP-induced conformational change is not mandatory for phosphorylation. As observed for the nucleotide-ATPase complex, the conformation of the first phosphorylated ATPase intermediate E1PCa(2) also depends on the nucleotide, indicating that ATPase states have a less uniform conformation than previously anticipated. PMID- 12538578 TI - The Mad and Myc basic domains are functionally equivalent. AB - The Myc/Max/Mad family of transcription factors plays a fundamental role in the regulation of cell proliferation, oncogenic transformation, and cell differentiation. However, it remains unclear whether different heterodimers, such as Myc/Max and Mad/Max, recognize the same or different target genes in vivo. We show by chromatin immunoprecipitation that Myc target genes are also recognized by Mad1 in differentiated HL60 cells. We also substituted the complete basic region of Myc for the corresponding region of Mad. Wild-type c-Myc was then compared with c-Myc(Mad-BR) in oncogenic transformation, regulation of cell proliferation, induction of apoptosis, activation of chromosomal gene expression, and direct binding to chromosomal sites by chromatin immunoprecipitation. We find that the wild-type c-Myc and c-Myc/MadBR proteins have indistinguishable biological activity and target gene recognition in vivo. These data are consistent with a model in which Myc and Mad regulate a common set of target genes. PMID- 12538579 TI - SC1/hevin. An extracellular calcium-modulated protein that binds collagen I. AB - SC1, a member of the BM-40 family of extracellular matrix proteins, was recombinantly expressed in a eukaryotic expression system. The full-length protein as well as truncated versions were purified to homogeneity under non denaturing conditions. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry of full-length SC1 revealed a mass of 87.8 kDa of which 16.8 kDa is contributed by posttranslational modifications. In electron microscopy, after negative staining, SC1 was revealed as a globule attached to a thread-like structure. A calcium dependence of the SC1 conformation could be demonstrated by fluorescence spectroscopy. In the extracellular matrix of cultured osteosarcoma cells SC1 was found associated with collagen I-containing fibrils, and binding of SC1 to reconstituted collagen I fibrils could be demonstrated by immunogold labeling and electron microscopy. SC1 showed a broad expression in a variety of tissues. PMID- 12538580 TI - Muscarinic receptor activation protects cells from apoptotic effects of DNA damage, oxidative stress, and mitochondrial inhibition. AB - The impact of muscarinic receptor stimulation was examined on apoptotic signaling induced by DNA damage, oxidative stress, and mitochondrial impairment. Exposure of human neuroblastoma SH-SY5Y cells to the DNA-damaging agent camptothecin increased p53 levels, activated caspase-3, and caused cell death. Pretreatment with oxotremorine-M, a selective agonist of muscarinic receptors that are expressed endogenously in these cells, did not affect the accumulation of p53 but greatly attenuated caspase-3 activation and protected from cell death to nearly the same extent as treatment with a general caspase inhibitor. Treatment with 50 200 microm H(2)O(2) caused the activation of caspase-3 beginning after 2-3 h, followed by eventual cell death. Oxotremorine-M pretreatment protected cells from H(2)O(2)-induced caspase-3 activation and death, and this was equivalent to protection afforded by a caspase inhibitor. Muscarinic receptor stimulation also protected cells from caspase-3 activation induced by exposure to rotenone, a mitochondrial complex 1 inhibitor, but no protection was evident from staurosporine-induced caspase-3 activation. The mechanism of protection afforded by muscarinic receptor activation from camptothecin-induced apoptotic signaling involved blockade of mitochondrial cytochrome c release associated with a bolstering of mitochondrial bcl-2 levels and blockade of the translocation of Bax to mitochondria. Likely the most proximal of these events to muscarinic receptor activation, mitochondrial Bax accumulation, also was attenuated by oxotremorine-M treatment after treatment with H(2)O(2) or rotenone. These results demonstrate that stimulation of muscarinic receptors provides substantial protection from DNA damage, oxidative stress, and mitochondrial impairment, insults that may be encountered by neurons in development, aging, or neurodegenerative diseases. These findings suggest that neurotransmitter-induced signaling bolsters survival mechanisms, and inadequate neurotransmission may exacerbate neuronal loss. PMID- 12538581 TI - Macrophage migration inhibitory factor deficiency is associated with altered cell growth and reduced susceptibility to Ras-mediated transformation. AB - Macrophage migration inhibitory factor (MIF) has been shown to functionally inactivate the p53 tumor suppressor and to inhibit p53-responsive gene expression and apoptosis. To better understand the role of MIF in cell growth and tumor biology, we evaluated MIF-null embryonic fibroblasts with respect to their immortalization and transformation properties. Although minor deviations in the growth characteristics of MIF(-/-) fibroblasts were observed under normal culture conditions, MIF-deficient cells were growth-impaired following the introduction of immortalizing oncogenes. The growth retardation by the immortalized MIF(-/-) cultures correlated with their reduced susceptibility to Ras-mediated transformation. Our results identify E2F as part of the restraining mechanism that is activated in response to oncogenic signaling and show that the biological consequences of E2F induction in MIF(-/-) fibroblasts vary depending on the p53 status, inducing predominantly G(1) arrest or apoptosis in p53-positive cells. This E2F activity is independent of Rb binding, but contingent on binding DNA. Resistance to oncogenic transformation by MIF(-/-) cells could be overcome by concomitant interference with p53- and E2F-responsive transcriptional control. Our results demonstrate that MIF plays a role in an E2F/p53 pathway that operates downstream of Rb regulation and implicate MIF as a mediator of normal and malignant cell growth. PMID- 12538583 TI - Benzyl-N-acetyl-alpha-D-galactosaminide induces a storage disease-like phenotype by perturbing the endocytic pathway. AB - The sugar analog O-benzyl-N-acetyl-alpha-d-galactosaminide (BG) is an inhibitor of glycan chain elongation and inhibits alpha2,3-sialylation in mucus-secreting HT-29 cells. Long-term exposure of these cells to BG is associated with the accumulation of apical glycoproteins in cytoplasmic vesicles. The mechanisms involved therein and the nature of the vesicles have not been elucidated. In these cells, a massive amount of BG metabolites is synthesized. Because sialic acid is mainly distributed apically in epithelial cells, it has been proposed that the BG-induced undersialylation of apical membrane glycoproteins is responsible for their intracellular accumulation due to a defect in anterograde traffic and that sialic acid may constitute an apical targeting signal. In this work, we demonstrate that the intracellular accumulation of membrane glycoproteins does not result mainly from defects in anterograde traffic. By contrast, in BG-treated cells, endocytosed membrane proteins were retained intracellularly for longer periods of time than in control cells and colocalized with accumulated MUC1 and beta(1) integrin in Rab7/lysobisphosphatidic acid(+) vesicles displaying features of late endosomes. The phenotype of BG-treated cells is reminiscent of that observed in lysosomal storage disorders. Sucrose induced a BG-like, lysosomal storage disease-like phenotype without affecting sialylation, indicating that undersialylation is not a requisite for the intracellular accumulation of membrane glycoproteins. Our findings strongly support the notion that the effects observed in BG-treated cells result from the accumulation of BG derived metabolites and from defects in the endosomal pathway. We propose that abnormal subcellular distribution of membrane glycoproteins involved in cellular communication and/or signaling may also take place in lysosomal storage disorders and may contribute to their pathogenesis. PMID- 12538582 TI - Interactions of a DNA-bound transcriptional activator with the TBP-TFIIA-TFIIB promoter quaternary complex. AB - Site-specific protein-DNA photo-cross-linking was used to show that, when bound to its cognate site at various distances upstream of the TATA element, the chimeric transcriptional activator GAL4-VP16 can physically interact with a TATA box-binding protein (TBP)- transcription factor IIA (TFIIA)-TFIIB complex assembled on the TATA element. This result implies DNA bending and looping of promoter DNA as a result of the physical interaction between GAL4-VP16 and an interface of the TBP-TFIIA-TFIIB complex. This protein-protein interaction on promoter DNA minimally requires the presence of one GAL4 binding site and the formation of a quaternary complex containing TBP, TFIIB, and TFIIA on the TATA element. Notably, the topology of the TBP-TFIIA-TFIIB-promoter complex is not altered significantly by the interaction with DNA-bound activators. We also show that the ability of GAL4-VP16 to activate transcription through a single GAL4 binding site varies according to its precise location and orientation relative to the TATA element and that it can approach the efficiency obtained with multiple binding sites. Taken together, our results indicate that the spatial positioning of the DNA-bound activation domain is important for efficient activation, possibly by maximizing its interactions with the transcriptional machinery including the TBP-TFIIA-TFIIB-promoter quaternary complex. PMID- 12538584 TI - Purification, characterization, molecular cloning, and expression of novel members of jacalin-related lectins from rhizomes of the true fern Phlebodium aureum (L) J. Smith (Polypodiaceae). AB - A lectin was purified from rhizomes of the fern Phlebodium aureum by affinity chromatography on mannose-Sepharose. The lectin, designated P. aureum lectin (PAL), is composed of two identical subunits of approximately 15 kDa associated by noncovalent bonds. From a cDNA library and synthetic oligonucleotide probes based on a partial amino acid sequence, 5'- and 3'-rapid amplification of cDNA ends allowed the generation of two similar full-length cDNAs, termed PALa and PALb, each of which had an open reading frame of 438 bp encoding 146 amino acid residues. The two proteins share 88% sequence identity and showed structural similarity to jacalin-related lectins. PALa contained peptide sequences exactly matching those found in the isolated lectin. PALa and PALb were expressed in Escherichia coli using pET-22b(+) vector and purified by one-step affinity chromatography. Native and recombinant forms of PAL agglutinated rabbit erythrocytes and precipitated with yeast mannan, dextran, and the high mannose containing glycoprotein invertase. The detailed carbohydrate-binding properties of the native and recombinant lectins were elucidated by agglutination inhibition assay, and native lectin was also studied by isothermal titration calorimetry. Based on the results of these assays, we conclude that this primitive vascular plant, like many higher plants, contains significant quantities of a mannose/glucose-binding protein in its storage tissue, whose binding specificity differs in detail from either legume mannose/glucose-binding lectins or monocot mannose-specific lectins. The identification of a jacalin-related lectin in a true fern reveals for the first time the widespread distribution and molecular evolution of this lectin family in the plant kingdom. PMID- 12538585 TI - Cell cycle modulation of gene targeting by a triple helix-forming oligonucleotide. AB - Successful gene-targeting reagents must be functional under physiological conditions and must bind chromosomal target sequences embedded in chromatin. Triple helix-forming oligonucleotides (TFOs) recognize and bind specific sequences via the major groove of duplex DNA and may have potential for gene targeting in vivo. We have constructed chemically modified, psoralen-linked TFOs that mediate site-specific mutagenesis of a chromosomal gene in living cells. Here we show that targeting efficiency is sensitive to the biology of the cell, specifically, cell cycle status. Targeted mutagenesis was variable across the cycle with the greatest activity in S phase. This was the result of differential TFO binding as measured by cross-link formation. Targeted cross-linking was low in quiescent cells but substantially enhanced in S phase cells with adducts in approximately 20-30% of target sequences. 75-80% of adducts were repaired faithfully, whereas the remaining adducts were converted into mutations (>5% mutation frequency). Clones with mutations could be recovered by direct screening of colonies chosen at random. These results demonstrate high frequency target binding and target mutagenesis by TFOs in living cells. Successful protocols for TFO-mediated manipulation of chromosomal sequences are likely to reflect a combination of appropriate oligonucleotide chemistry and manipulation of the cell biology. PMID- 12538586 TI - Lipid rafts/caveolae are essential for insulin-like growth factor-1 receptor signaling during 3T3-L1 preadipocyte differentiation induction. AB - Lipid rafts/caveolae are found to be essential for insulin-like growth factor (IGF)-1 receptor signaling during 3T3-L1 preadipocyte differentiation induction. In 3T3-L1 cells, IGF-1 receptor is located in lipid rafts/caveolae of the plasma membrane and can directly interact with caveolin-1, the major protein component in caveolae. Disruption of lipid rafts/caveolae by depleting cellular cholesterol with cholesterol-binding reagent, beta-methylcyclodextrin or filipin, blocks the IGF-1 receptor signaling in 3T3-L1 preadipocyte. Both hormonal induced adipocyte differentiation and mitotic clonal expansion are inhibited by lipid rafts/caveolae disruption. However, a nonspecific lipid binding reagent, xylazine, does not affect adipocyte differentiation or mitotic clonal expansion. Further studies indicate that lipid rafts/caveolae are required only for IGF-1 receptor downstream signaling and not the activation of receptor itself by ligand. Thus, our results suggest that localization in lipid rafts/caveolae and association with caveolin enable IGF-1 receptor to have a close contact with downstream signal molecules recruited into lipid rafts/caveolae and transmit the signal through these signal molecule complexes. PMID- 12538587 TI - The modulation of tissue factor by endothelial cells during heat shock. AB - Tissue factor (TF) initiates the extrinsic coagulation cascade on the surface of macrophages and endothelial cells. In septic patients, the extrinsic coagulation cascade is activated. When septic patients are febrile, mortality is decreased. The purpose of this study was to investigate the role of elevated temperatures on TF expression by endothelial cells during a sepsis-like challenge. Human endothelial vein cells (HUVECs) were incubated with lipopolysaccharide (LPS) or interleukin-1 beta (IL-1 beta) for 0, 2, 4, 6, or 8 h. At the 0-h time point, some HUVECs were heat shocked at 43 degrees C for 2 h and then recovered at 37 degrees C for 0, 2, 4, or 6 h. Heat-shocked and non-heat-shocked LPS-stimulated HUVECs were analyzed for TF-specific mRNA expression by ribonuclease protection assay (RPA), surface TF expression by flow cytometry, and TF activity by a two stage clotting assay. Heat shocked LPS-stimulated HUVECs expressed significantly reduced TF-specific mRNA, TF surface protein levels, and TF surface activity when compared with non-heat-shocked, LPS-stimulated HUVECs (p < 0.0125, p < 0.0125, and p < 0.0001, respectively; repeated measures analysis of variance, ANOVA). If heat shock models elevated core temperature, these results suggest that fever may protect the host during sepsis by reducing TF activity on the surface of endothelial cells. PMID- 12538588 TI - Kruppel-like factor 4 (KLF4/GKLF) is a target of bone morphogenetic proteins and transforming growth factor beta 1 in the regulation of vascular smooth muscle cell phenotype. AB - Vascular smooth muscle cell (VSMC) differentiation and phenotypic modulation is characterized by changes in mRNA expression for smooth muscle (SM) marker contractile proteins such as alpha-SM actin and SM22 alpha. Transforming growth factor beta1 (TGF-beta 1) is a potent VSMC differentiation factor; however, it is not known if other TGF-beta-superfamily members, in particular the bone morphogenetic proteins (BMPs), modulate VSMC phenotype. Here we demonstrate that a large subset of TGF-beta-superfamily members and their type I receptors are differentially co-expressed as VSMC phenotype changes during fetal/neonatal development and that BMP2, -4, and -6 reciprocally regulate SM-marker mRNA and protein expression in vitro. BMP2 and BMP6 decrease expression of the SM markers alpha-SM actin, SM22alpha, and calponin in rat VSMCs, whereas BMP4 increases their expression. The effects of BMP-2, -4, and -6 on SM marker gene transcription are mediated through a consensus TGF-beta-controlling element, the TCE, which is common to regulatory regions of SM-marker genes. Moreover, co treatment experiments revealed that BMP-2, -4, and -6 each inhibit TGF-beta 1 modulated increases in SM22alpha reporter gene activity. Regardless of whether they positively or negatively regulate SM marker expression, TGF-beta 1 and BMP 2, -4, and -6 all induced binding of the Kruppel-like transcription factor, GKLF/KLF4, to the TGF-beta control element. Induction of KLF4 was confirmed by immunocytochemistry and Western blotting, which revealed that a lower molecular weight KLF4 protein is induced after treatment with TGF-beta-superfamily members. Taken together, our results demonstrate that multiple members of the TGF-beta superfamily act in concert to modulate VSMC phenotype. PMID- 12538589 TI - Regulation of a transient receptor potential (TRP) channel by tyrosine phosphorylation. SRC family kinase-dependent tyrosine phosphorylation of TRPV4 on TYR-253 mediates its response to hypotonic stress. AB - The recently identified transient receptor potential (TRP) channel family member, TRPV4 (formerly known as OTRPC4, VR-OAC, TRP12, and VRL-2) is activated by hypotonicity. It is highly expressed in the kidney as well as blood-brain barrier deficient hypothalamic nuclei responsible for systemic osmosensing. Apart from its gating by hypotonicity, little is known about TRPV4 regulation. We observed that hypotonic stress resulted in rapid tyrosine phosphorylation of TRPV4 in a heterologous expression model and in native murine distal convoluted tubule cells in culture. This tyrosine phosphorylation was sensitive to the inhibitor of Src family tyrosine kinases, PP1, in a dose-dependent fashion. TRPV4 associated with Src family kinases by co-immunoprecipitation studies and confocal immunofluorescence microscopy, and this interaction required an intact Src family kinase SH2 domain. One of these kinases, Lyn, was activated by hypotonic stress and phosphorylated TRPV4 in an immune complex kinase assay and an in vitro kinase assay using recombinant Lyn and TRPV4. Transfection of wild-type Lyn dramatically potentiated hypotonicity-dependent TRPV4 tyrosine phosphorylation whereas dominant negative-acting Lyn modestly inhibited it. Through mutagenesis studies, the site of tonicity-dependent tyrosine phosphorylation was mapped to Tyr-253, which is conserved across all species from which TRPV4 has been cloned. Importantly, point mutation of Tyr-253 abolished hypotonicity-dependent channel activity. In aggregate, these data indicate that hypotonic stress results in Src family tyrosine kinase-dependent tyrosine phosphorylation of the tonicity sensor TRPV4 at residue Tyr-253 and that this residue is essential for channel function in this context. This is the first example of direct regulation of TRP channel function through tyrosine phosphorylation. PMID- 12538590 TI - Proteomic analysis of calcium/calmodulin-dependent protein kinase I and IV in vitro substrates reveals distinct catalytic preferences. AB - The multifunctional calcium/calmodulin-dependent protein kinases I and IV (CaMKI and CaMKIV) are closely related by primary sequence and predicted to have similar substrate specificities based on peptide studies. We identified a fragment of p300-(1-117) that is a substrate of both kinases, and through both mutagenesis and Edman phosphate ((32)P) release sequencing, established that CaMKI and CaMKIV phosphorylate completely different sites. The CaMKI site, Ser(89) ((84)LLRSGSSPNL(93)), fits the expected consensus whereas the CaMKIV site, Ser(24) ((19)SSPALSASAS(28)), is novel. To compare kinase substrate preferences more generally, we employed a proteomic display technique that allowed comparison of complex cell extracts phosphorylated by each kinase in a rapid in vitro assay, thereby demonstrating substrate preferences that overlapped but were clearly distinct. To validate this approach, one of the proteins labeled in this assay was identified by microsequencing as HSP25, purified as a recombinant protein, and examined as a substrate for both CaMKI and CaMKIV. Again, CaMKI and CaMKIV were different, this time in kinetics and stoichiometry of the phosphorylation sites, with CaMKI preferring Ser(15) ((10)LLRTPSWGPF(19)) to Ser(85) ((80)LNRQLSSGVS(89)) 3:1, but CaMKIV phosphorylating the two sites equally. These differences in substrate specificities emphasize the need to consider these protein kinases independently despite their close homology. PMID- 12538591 TI - Ryanodine receptor type I and nicotinic acid adenine dinucleotide phosphate receptors mediate Ca2+ release from insulin-containing vesicles in living pancreatic beta-cells (MIN6). AB - We have demonstrated recently (Mitchell, K. J., Pinton, P., Varadi, A., Tacchetti, C., Ainscow, E. K., Pozzan, T., Rizzuto, R., and Rutter, G. A. (2001) J. Cell Biol. 155, 41-51) that ryanodine receptors (RyR) are present on insulin containing secretory vesicles. Here we show that pancreatic islets and derived beta-cell lines express type I and II, but not type III, RyRs. Purified by subcellular fractionation and membrane immuno-isolation, dense core secretory vesicles were found to possess a similar level of type I RyR immunoreactivity as Golgi/endoplasmic reticulum (ER) membranes but substantially less RyR II than the latter. Monitored in cells expressing appropriately targeted aequorins, dantrolene, an inhibitor of RyR I channels, elevated free Ca(2+) concentrations in the secretory vesicle compartment from 40.1 +/- 6.7 to 90.4 +/- 14.8 microm (n = 4, p < 0.01), while having no effect on ER Ca(2+) concentrations. Furthermore, nicotinic acid adenine dinucleotide phosphate (NAADP), a novel Ca(2+)-mobilizing agent, decreased dense core secretory vesicle but not ER free Ca(2+) concentrations in permeabilized MIN6 beta-cells, and flash photolysis of caged NAADP released Ca(2+) from a thapsigargin-insensitive Ca(2+) store in single MIN6 cells. Because dantrolene strongly inhibited glucose-stimulated insulin secretion (from 3.07 +/- 0.51-fold stimulation to no significant glucose effect; n = 3, p < 0.01), we conclude that RyR I-mediated Ca(2+)-induced Ca(2+) release from secretory vesicles, possibly potentiated by NAADP, is essential for the activation of insulin secretion. PMID- 12538592 TI - Phosphorylation of the Ras-GRF1 exchange factor at Ser916/898 reveals activation of Ras signaling in the cerebral cortex. AB - The Ras-GRF1 exchange factor, which is regulated by increases in intracellular calcium and the release of G beta gamma subunits from heterotrimeric G proteins, plays a critical role in the activation of neuronal Ras. Activation of G protein coupled receptors stimulates an increase in the phosphorylation of Ras-GRF1 at certain serine residues. The first of these sites to be identified, Ser(916) in the mouse sequence (equivalent to Ser(898) in the rat sequence), is required for full activation of the Ras exchange factor activity of Ras-GRF1 by muscarinic receptors. We demonstrate here that Ras-GRF1 is highly expressed in rat brain compared with the Sos exchange factor and that there is an increase in incorporation of (32)P into Ser(898) of brain Ras-GRF1 following activation of protein kinase A. Phosphorylation of Ras-GRF1 at Ser(916) is also required for maximal induction of Ras-dependent neurite outgrowth in PC12 cells. A novel antibody (termed 2152) that selectively recognizes Ras-GRF1 when it is phosphorylated at Ser(916/898) confirmed the regulated phosphorylation of Ras GRF1 by Western blotting in both model systems of transfected COS-7 and PC12 cells and also of the endogenous protein in rat forebrain slices. Indirect confocal immunofluorescence of transfected PC12 cells using antibody 2152 demonstrated reactivity only under conditions in which Ras-GRF1 was phosphorylated at Ser(916/898). Confocal immunofluorescence of cortical slices of rat brain revealed widespread and selective phosphorylation of Ras-GRF1 at Ser(898). In the prefrontal cortex, there was striking phosphorylation of Ras GRF1 in the dendritic tree, supporting a role for Ras activation and signal transduction in neurotransmission in this area. PMID- 12538593 TI - Alternative mRNAs arising from trans-splicing code for mitochondrial and cytosolic variants of Echinococcus granulosus thioredoxin Glutathione reductase. AB - Thioredoxin and glutathione systems are the major thiol-dependent redox systems in animal cells. They transfer via the reversible oxidoreduction of thiols the reducing equivalents of NADPH to numerous substrates and substrate reductases and constitute major defenses against oxidative stress. In this study, we cloned from the helminth parasite Echinococcus granulosus two trans-spliced mRNA variants that encode thioredoxin glutathione reductases (TGR). These variants code for mitochondrial and cytosolic selenocysteine-containing isoforms that possess identical glutaredoxin (Grx) and thioredoxin reductase (TR) domains and differ exclusively in their N termini. Western blot analysis of subcellular fractions with specific anti-TGR antibodies showed that TGR is present in both compartments. The biochemical characterization of the native purified TGR suggests that the Grx and TR domains of the enzyme can function either coupled or independently of each other, because the Grx domain can accept electrons from either TR domains or the glutathione system and the TR domains can transfer electrons to either the fused Grx domain or to E. granulosus thioredoxin. PMID- 12538594 TI - Characterization of the Xenopus galectin family. Three structurally different types as in mammals and regulated expression during embryogenesis. AB - We have isolated six novel galectin cDNAs from a Xenopus laevis kidney cDNA library. The newly identified X. laevis galectins (xgalectins) comprise one proto type (xgalectin-Vb), one chimera type (xgalectin-VIIa), and four tandem repeat types (xgalectin-IIb, -IIIb, -VIa, and -VIIIa). Thus, together with those mentioned in our previous work (Shoji, H., Nishi, N., Hirashima, M., and Nakamura, T. (2002) Glycobiology 12, 163-172), the 12 xgalectins are classified into three types based on their domain structures, as in mammals. The xgalectins whose counterparts in other species have not been identified (xgalectin-IVa, -Vb, and -VIa) were confirmed to possess lactose-binding activity by expression of their recombinant forms. This shows that they truly function as animal lectins. The protein purification study revealed that the major xgalectins in kidney are xgalectin-Ib, -IIa, -IIb, -IIIa, and -VIIa. The mRNAs of xgalectin-IIb, -IIIb, Vb, and -VIa were localized to specific adult tissues, whereas those of xgalectin VIIa and -VIIIa were broadly distributed. The temporal expression patterns of the mRNAs of the 12 xgalectins during embryogenesis were analyzed and categorized into three groups: 1) mRNA observed to exist throughout embryogenesis, i.e. maternal mRNA also exists (xgalectin-Ia, -IIa, -IIIa, -IIIb, -Va, -VIIa, and VIIIa); 2) mRNA observed from the gastrula stage (xgalectin-VIa); and 3) mRNA observed from the tail bud or the tadpole stage (xgalectin-Ib, -IIb, -IVa, and Vb). The mRNA of the most abundant xgalectin in embryos, xgalectin-VIIa, was localized to the surface layer of embryos, the epidermis, the cement gland, and various placodes. Xgalectin-VIIa protein was also observed to exist throughout embryogenesis by Western blot analysis with specific antiserum. These results show that the expression of each member is spatiotemporally regulated from eggs to adulthood, suggesting that galectins play multiple roles not only in adults, but also in development. PMID- 12538595 TI - Activation of the mitogen-activated protein kinases Erk1/2 by erythropoietin receptor via a G(i )protein beta gamma-subunit-initiated pathway. AB - We have recently shown that a heterotrimeric G(i) protein is coupled to the erythropoietin (Epo) receptor. The G(i) protein constitutively associates in its heterotrimeric form with the intracellular domain of Epo receptor (EpoR). After Epo stimulation G(i) is released from the receptor and activated. In the present study we have investigated the functional role of the heterotrimeric G(i) protein bound to EpoR. In Chinese hamster ovary cells expressing EpoR, the G(i) inhibitor pertussis toxin blocked mitogen-activated protein kinase (MAPK) Erk1/2 activation induced by Epo. Epo-dependent MAPK activation was also sensitive to the G beta gamma competitive inhibitor beta ARK1-ct (C-terminal fragment of the beta adrenergic receptor kinase), to the Ras dominant negative mutant RasN17, and to the phosphoinositide 3-kinase (PI3K) inhibitor LY 294002. A region of 7 amino acids (469-475) in the C-terminal end of EpoR was shown to be required for G(i) binding to EpoR in vivo. Deletion of this region in EpoR abolished both MAPK and PI3K activation in response to Epo. We conclude that in Chinese hamster ovary cells, Epo activates MAPK via a novel pathway dependent on G(i) association to EpoR, G beta gamma subunit, Ras, and PI3K. The tyrosine kinase Jak2 also contributes to this new pathway, more likely downstream of beta gamma and upstream of Ras and PI3K. This pathway is similar to the best characterized pathway used by seven transmembrane receptors coupled to G(i) to activate MAPK and may cooperate with other described Epo-dependent MAPK activation pathways in hematopoietic cells. PMID- 12538596 TI - Subcellular localization of beta-arrestins is determined by their intact N domain and the nuclear export signal at the C terminus. AB - beta-Arrestin1 and beta-arrestin2 play a key role in the regulation of G protein coupled receptor-mediated signaling, whereas the subcellular distribution of beta arrestin1 and beta-arrestin2 has been shown to be quite different. In this study, we found that although both beta-arrestin1 and beta-arrestin2 are able to interact with ubiquitin-protein isopeptide ligase (E3) Mdm2, only expression of beta-arrestin2 leads to the relocalization of Mdm2 from the nucleus to the cytoplasm. Further study reveals that beta-arrestin2 but not beta-arrestin1 shuttles between the cytoplasm and nucleus in a leptomycin B-sensitive manner. A hydrophobic amino acid-rich region (VXXXFXXLXL) at the C terminus of beta arrestin2 was further demonstrated to serve as a nuclear export signal responsible for the extranuclear localization of beta-arrestin2. In the corresponding region of beta-arrestin1, there is a single amino acid difference (Glu instead of Leu in beta-arrestin2), and mutation of Glu to Leu conferred to beta-arrestin1 similar subcellular distribution to that of beta-arrestin2. Moreover, data from a series of deletion mutations demonstrated that the N domain (residues 1-185) was indispensable for the nuclear localization of both beta arrestins, and the results from a Val to Asp point mutation in the N domain also supported this notion. In addition, our data showed that nucleocytoplasmic shuttling of beta-arrestin2 was required, via protein/protein interaction, for the cytoplasmic relocalization of Mdm2 and JNK3, another well known beta arrestin2-binding protein. Our study thus suggests that both the nuclear export signal motif and the N domain of beta-arrestins are critical for the regulation of their subcellular localization and that beta-arrestin2 may modulate the function of its binding partners such as Mdm2 and JNK3 by alteration of their subcellular distribution. PMID- 12538597 TI - Regional loss of the mitochondrial membrane potential in the hepatocyte is rapidly followed by externalization of phosphatidylserines at that specific site during apoptosis. AB - The spatio-temporal relationship between a decrease in the mitochondrial membrane potential (MMP) and externalization of phosphatidylserines (PS) during induction of apoptosis was investigated in single freshly isolated hepatocytes. Apoptosis was induced in the hepatocytes in three different ways: attack by activated Natural Killer cells, exposure to ATP, or exposure to the inhibitor of protein synthesis cycloheximide. Fluorescence microscopy showed staining of externalized PS at those areas where the staining for MMP was lost whereas in other areas the mitochondria remained intact for longer periods of time, indicating coupling between local loss of MMP and local PS exposure. To discriminate whether the decrease in MMP itself or a decrease in ATP induced PS externalization, hepatocytes were treated with rotenone, which resulted in a rapid collapse of cellular ATP but left the MMP intact for a much longer period. Addition of fructose prevented the decrease of ATP to approximately 30% and also delayed the collapse of the MMP. This indicates that ATP was needed for the maintenance of the MMP probably via reverse action of the ATP synthase. In a subsequent study hepatocytes were incubated with Natural Killer cells for induction of apoptosis followed by addition of rotenone to deplete ATP. Under these conditions the PS staining co-localized with mitochondrial MMP indicating that PS externalization does not require a collapse in MMP. Moreover, exposure of PS was evenly distributed over the whole plasma membrane. In conclusion, we propose that after an apoptotic stimulus some mitochondria start to loose their MMP, which results in cessation of ATP production and perhaps even consumption of ATP. This results in an overall decrease in cellular ATP. ATP-consuming enzyme reactions most distal from still intact mitochondria will be most sensitive to such a decrease. Apparently the translocase that keeps phosphatidylserines inward-oriented is such a sensitive enzyme. PMID- 12538598 TI - Neutralization of circulating vascular endothelial growth factor (VEGF) by anti VEGF antibodies and soluble VEGF receptor 1 (sFlt-1) induces proteinuria. AB - There are about 2.5 million glomeruli in the kidneys each consisting of a barrel of glomerular basement membrane surrounded by glomerular endothelial cells on the inside and glomerular epithelial cells with established foot processes (podocytes) on the outside. Defects in this filtration apparatus lead to glomerular vascular leak or proteinuria. The role of vascular endothelial growth factor (VEGF) in the regulation of glomerular vascular permeability is still unclear. Recent studies indicate that patients receiving anti-VEGF antibody therapy may have an increased incidence of proteinuria. In a different setting, pregnancies complicated by preeclampsia are associated with elevated soluble VEGF receptor 1 protein (sFlt-1), endothelial cell dysfunction and proteinuria. These studies suggest that neutralization of physiologic levels of VEGF, a key endothelial survival factor, may lead to proteinuria. In the present study, we evaluated the potential of anti-VEGF neutralizing antibodies and sFlt-1 in the induction of proteinuria. Our studies demonstrate that anti-VEGF antibodies and sFlt-1 cause rapid glomerular endothelial cell detachment and hypertrophy, in association with down-regulation of nephrin, a key epithelial protein in the glomerular filtration apparatus. These studies suggest that down-regulation or neutralization of circulating VEGF may play an important role in the induction of proteinuria in various kidney diseases, some forms of cancer therapy and also in women with preeclampsia. PMID- 12538599 TI - Minireview: parathyroid hormone-related protein as an intracrine factor- trafficking mechanisms and functional consequences. AB - PTH-related protein (PTHrP) was originally discovered as the factor responsible for humoral hypercalcemia of malignancy. PTHrP is produced by most cell types and is a prohormone that gives rise to a family of mature secretory forms arising from posttranslational endoproteolytic cleavage of the initial translation product. Each of these secretory forms of PTHrP is believed to have one or more of its own receptors on the cell surface that mediates the normal paracrine, autocrine, and endocrine actions of PTHrP. Recently, evidence has accumulated that indicates that PTHrP is also able to enter the nucleus and/or the nucleolus and influence cellular events in an intracrine fashion. This review discusses the mechanisms by which PTHrP may gain access to the nucleus/nucleolus and the functional consequences of this nuclear entry by PTHrP. PMID- 12538601 TI - A transcriptionally active human type II gonadotropin-releasing hormone receptor gene homolog overlaps two genes in the antisense orientation on chromosome 1q.12. AB - GnRH-II peptide hormone exhibits complete sequence conservation across vertebrate species, including man. Type-II GnRH receptor genes have been characterized recently in nonhuman primates, but the human receptor gene homolog contains a frameshift, a premature stop codon (UGA), and a 3' overlap of the RBM8A gene on chromosome 1q.12. A retrotransposed pseudogene, RBM8B, retains partial receptor sequence. In this study, bioinformatics show that the human receptor gene promoter overlaps the peroxisomal protein 11-beta gene promoter and the premature UGA is positionally conserved in chimpanzee. A CGA [arginine (Arg)] occurs in porcine DNA, but UGA is shifted one codon to the 5' direction in bovine DNA, suggesting independent evolution of premature stop codons. In contrast to marmoset tissue RNA, exon- and strand-specific probes are required to distinguish differently spliced human receptor gene transcripts in cell lines (HP75, IMR-32). RBM8B is not transcribed. Sequencing of cDNAs for spliced receptor mRNAs showed no evidence for alteration of the premature UGA by RNA editing, but alternative splicing circumvents the frameshift to encode a two-membrane-domain protein before this UGA. A stem-loop motif resembling a selenocysteine insertion sequence and a potential alternative translation initiation site might enable expression of further proteins involved in interactions within the GnRH system. PMID- 12538602 TI - Selection of the dominant follicle and insulin-like growth factor (IGF)-binding proteins: evidence that pregnancy-associated plasma protein A contributes to proteolysis of IGF-binding protein 5 in bovine follicular fluid. AB - Development of a dominant follicle is associated with decreased intrafollicular low molecular weight IGF-binding proteins (namely IGFBP-2, -4, and -5) and increased proteolysis of IGFBP-4 by pregnancy-associated plasma protein A (PAPP A). In addition to IGFBP-4 proteolytic activity, bovine follicular fluid contains strong proteolytic activity for IGFBP-5, but not for IGFBP-2. Here we show that the IGFBP-5 protease present in bovine follicular fluid is a neutral/basic pH favoring, Zn(2+) metalloprotease very similar to the previously described IGFBP-4 protease. We hypothesized that immunoneutralization and immunoprecipitation with anti-PAPP-A antibodies would result in abrogation of the IGFBP-4, but not the IGFBP-5, proteolytic activity in follicular fluid. As expected, anti-PAPP-A antibodies were able to neutralize and precipitate the IGFBP-4, but not the IGFBP 5, proteolytic activity of human pregnancy serum, which was used as a positive control for PAPP-A. Surprisingly, immunoneutralization and immunoprecipitation of follicular fluid from bovine preovulatory follicles with anti-PAPP-A antibodies abrogated both IGFBP-4 and IGFBP-5 proteolysis. Quantitative results derived from phosphorimaging revealed a complete inhibition of both IGFBP-4 and -5 proteolysis by follicular fluid incubated for 2 or 5 h in the presence of anti-PAPP-A antibodies. After 18 h of incubation, anti-PAPP-A antibodies still inhibited IGFBP-5 degradation, although with an efficiency lower than that for IGFBP-4 degradation. Both proteolytic activities have identical electrophoretic mobility, and a single band ( approximately 400 kDa) was detected by Western immunoblotting of bovine follicular fluid with anti-PAPP-A antibodies. Proteolysis of IGFBP-5 was readily detectable in follicular fluid from dominant follicles and was negligible in subordinate follicles from the same cohort. These results suggest that an active intrafollicular IGFBP-4/-5 proteolytic system, in which PAPP-A is the major protease involved, is an important determinant of follicular fate. PMID- 12538600 TI - Glucocorticoids induce rapid up-regulation of mitogen-activated protein kinase phosphatase-1 and dephosphorylation of extracellular signal-regulated kinase and impair proliferation in human and mouse osteoblast cell lines. AB - A central feature of glucocorticoid (GC)-induced osteoporosis is decreased bone formation, secondary to decreased numbers of functional osteoblasts. We find that ERK activity is essential for serum-induced osteoblast proliferation in vitro because inhibition of MAPK/ERK kinase activity by U0126 completely abolished both serum-induced activation of ERK and proliferation of mouse (MBA-15.4) and human (MG-63) osteoblast cell lines. Dexamethasone (Dex) rapidly (<2 h) inhibits the sustained phase of ERK activation, required for nuclear shift and mitogenesis. This inhibition is reversed by cotreatment with the protein synthesis inhibitor, cycloheximide, and by the GC receptor antagonist, RU486, suggesting a classical transcriptional mechanism. Phosphatase activity was up-regulated by Dex treatment, and inhibition of ERK activity by Dex was also reversed by the protein tyrosine phosphatase inhibitor, vanadate. Coupled with the rapidity of Dex action, this indicates immediate-early gene phosphatase involvement, and we therefore used quantitative, real-time PCR to examine expression profiles of the dual-specificity MAPK phosphatases, MKP-1 and MKP-3. MKP-1, but not MKP-3, mRNA expression was 10-fold up-regulated in both mouse and human osteoblast cell lines within 30 min of Dex treatment and remained elevated for 24 h. MKP-1 protein was also markedly up-regulated following 1-8 h of Dex treatment, and this correlated precisely with dephosphorylation of ERK. Cell proliferation was impaired by Dex treatment, and this was reversed by both RU486 and vanadate. Therefore, MKP-1 up regulation provides a novel and rapid mechanism, whereby GCs inhibit osteoblast proliferation. PMID- 12538603 TI - Coexpression of calcitonin receptor-like receptor and receptor activity-modifying protein 2 or 3 mediates the antimigratory effect of adrenomedullin. AB - Three isoforms of the receptor activity-modifying protein (RAMP) are thought to transport the calcitonin receptor-like receptor (CRLR) to the plasma membrane to function as calcitonin gene-related peptide or adrenomedullin receptors, but their role remains largely unknown. We investigated whether coexpression of RAMP and CRLR are involved in the regulation of cell migration using a monolayer wounding protocol. Quantification of gene transcripts revealed expression of all RAMP isoforms and CRLR in cultured rat vascular smooth muscle cells (VSMCs), RAMP2 and RAMP3 in rat endothelial cells, and RAMP1 in rat fibroblasts. CRLR expression was minimal in endothelial cells and fibroblasts. Adrenomedullin potently suppressed the migration of VSMCs, whereas calcitonin gene-related peptide did not suppress migration in any cell type. The antimigratory effect of adrenomedullin on VSMCs was potentiated by transfecting CRLR cDNA. Cotransfection of RAMP2 or RAMP3 with CRLR into VSMCs resulted in a slower migratory rate, and this effect was enhanced by adrenomedullin. Migration of fibroblasts was also suppressed after cotransfection of RAMP2 or RAMP3 with CRLR. cAMP agonists had no effect on VSMC migration, and a cAMP antagonist failed to abrogate the antimigratory effect of adrenomedullin. Thus, coexpression of CRLR and RAMP2 or RAMP3 mediates the inhibitory effect of adrenomedullin on cell migration, independent of cAMP-dependent signaling pathways. PMID- 12538604 TI - Ala/Thr(201) in extracellular loop 2 and Leu/Phe(290) in transmembrane domain 6 of type 1 frog gonadotropin-releasing hormone receptor confer differential ligand sensitivity and signal transduction. AB - Recently, we have identified three distinct types of bullfrog GnRH receptor (designated bfGnRHR-1, bfGnRHR-2, and bfGnRHR-3). In the present study, we have isolated three GnRHR clones in Rana dybowskii (dyGnRHR-1, dyGnRHR-2, and dyGnRHR 3). Despite high homology of dyGnRHRs with the corresponding bfGnRHRs, dyGnRHRs revealed different signaling pathways and ligand sensitivity compared with the bfGnRHR counterparts. Activation of dyGnRHRs with GnRH stimulated cAMP-mediated gene expression. However, dyGnRHR-3 but not dyGnRHR-1 and -2 induced c-fos promoter-driven gene expression. Consistently, dyGnRHR-1 and dyGnRHR-2 were not able to increase GnRH-induced inositol phosphate accumulation, whereas all bfGnRHRs and dyGnRHR-3 were, indicating that dyGnRHR-1 and dyGnRHR-2 are coupled to solely G(s), whereas all bfGnRHRs and dyGnRHR-3 are coupled to both G(s) and G(q/11). Moreover, dyGnRHR-1 and dyGnRHR-2 showed about 10-fold less sensitivity to each ligand than that of the bfGnRHR counterparts. Using type 1 chimeric and point-mutated receptors, we further elucidated that specific amino acids, Ala/Thr(201) in extracellular loop 2 and Leu/Phe(290) in transmembrane domain 6 of the type 1 receptor, are responsible for ligand sensitivity and signal transduction pathway. Particularly, substitution of Leu(290) to Phe in dyGnRHR-1 increased GnRH-induced inositol phosphate production as well as c-fos promoter driven gene expression whereas substitution of Phe(290) to Leu in bfGnRHR-1 decreased those activities. Collectively, these results demonstrate the presence of three types of GnRHR in amphibians, and suggest species- and type-specific ligand recognition and different signaling pathways in frog GnRHRs. PMID- 12538606 TI - Molecular target of endocrine disruption in human luteinizing granulosa cells by 2,3,7,8-tetrachlorodibenzo-p-dioxin: inhibition of estradiol secretion due to decreased 17alpha-hydroxylase/17,20-lyase cytochrome P450 expression. AB - Estradiol (E2) production by human luteinized granulosa cells (hLGC) is inhibited by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The molecular target of TCDD toxicity has not been identified. The decrease in E2 is ameliorated by androgen substrate addition and is not associated with changes in aromatase cytochrome P450 (P450arom) activity or protein expression. An antihuman 17alpha hydroxylase/17,20-lyase cytochrome P450 (P450c17) antisera and a direct radiometric assay of 17,20-lyase activity were used to test the hypothesis that TCDD targets P450c17, thereby decreasing substrate availability for E2 synthesis by hLGC. P450c17 expression and 17,20-lyase activity were detected in hLGC with high levels of E2 secretion. Western immunoblot analysis demonstrated that TCDD treatment of hLGC decreased the expression of P450c17 by as much 50% (P < 0.05). TCDD exposure induced a 65% decrease in 17,20-lyase activity (P < 0.05), but no changes were seen in P450arom or in nicotinamide adenine dinucleotide phosphate (reduced)-cytochrome P450 oxidoreductase (reductase). Furthermore, the decreases in P450c17 and 17,20-lyase were proportional to the inhibition of E2 secretion. We conclude that the molecular target for endocrine disruption of hLGC by TCDD is P450c17, specifically decreasing the supply of androgens for E2 synthesis, and that it does not involve either P450arom or the redox partner protein reductase. PMID- 12538607 TI - Proliferative phase sertoli cells display a developmentally regulated response to activin in vitro. AB - We have used cultures of highly purified, proliferating rat Sertoli cells collected from d 3, 6, and 9 rat pups to investigate the role of activin A on Sertoli cell division. These studies demonstrate that activin A acts directly on d 6 and 9, but not d 3, Sertoli cells to induce proliferation, both alone and synergistically with FSH. In addition to stimulating proliferation, activin A induces secretion of inhibins A and B as determined by specific ELISAs. We demonstrate that the synergy between activin A and FSH is not due to local actions of secreted inhibin or follistatin. We have used real-time fluorometric RT-PCR to demonstrate that activin regulates expression of activin receptor and follistatin mRNA by Sertoli cells. Saturation binding studies using (125)I activin A indicate that synergy between activin and FSH may be due to increased numbers of activin receptors on the Sertoli cell. Finally, we show that activin A was secreted at high levels by cultured peritubular cells but was undetectable in high purity proliferating Sertoli cell cultures, suggesting that activin A functions as a paracrine factor during postnatal testis development. PMID- 12538608 TI - Galanin enhancement of gonadotropin-releasing hormone-stimulated luteinizing hormone secretion in female rats is estrogen dependent. AB - The hypothalamic peptide GnRH is the primary neuroendocrine signal regulating pituitary LH in females. The neuropeptide galanin is cosecreted with GnRH from hypothalamic neurons, and in vitro studies have demonstrated that galanin can act at the level of the pituitary to directly stimulate LH secretion and also augment GnRH-stimulated LH secretion. Several lines of evidence have suggested that the hypophysiotropic effects of galanin are important for the generation of preovulatory LH surges. To determine whether the pituitary actions of galanin are enhanced by the preovulatory steroidal milieu, LH responses to galanin administration (with or without GnRH) were examined in: 1) ovariectomized (OVX); 2) OVX, estrogen (E)-primed; and 3) OVX, E- and progesterone-treated female rats. Results from the study indicate that galanin enhances GnRH-stimulated LH secretion only in the presence of E (in OVX, E-primed, or E- and progesterone treated rats). Galanin alone does not directly stimulate LH secretion under any of the steroid conditions examined. In the absence of gonadal steroids (OVX rats), galanin inhibits GnRH-stimulated LH secretion. These findings suggest that the primary pituitary effect of galanin is to modulate GnRH-stimulated LH secretion, and that the potentiating effects of galanin occur only in the presence of E. PMID- 12538609 TI - Involvement of cyclic adenosine 3',5'-monophosphate in the differential regulation of activin betaA and betaB expression by gonadotropin in the zebrafish ovarian follicle cells. AB - Activin is a dimeric protein consisting of two similar but distinct beta subunits, betaA and betaB. In our previous studies, both activin A (betaAbetaA) and activin B (betaBbetaB) have been demonstrated to stimulate oocyte maturation and promote oocyte maturational competence in the zebrafish. Follistatin, a specific activin-binding protein, can block both activin- and gonadotropin induced final oocyte maturation in vitro, suggesting that activin is likely a downstream mediator of gonadotropin actions in the zebrafish ovary. In the present study, a full-length cDNA encoding zebrafish ovarian activin betaA was cloned and sequenced. The precursor of zebrafish activin betaA consists of 395 amino acids and its mature region exhibits about 78% homology with that of mammals. Using an in vitro primary culture of the ovarian follicle cells and semiquantitative RT-PCR assays, we examined the regulation of activin betaA and betaB expression by human chorionic gonadotropin (hCG) and its intracellular signal transduction mechanisms. hCG (15 IU/ml) increased the mRNA level of activin betaA-subunit; however, it significantly down-regulated the steady-state expression level of activin betaB in a time- and dose-dependent manner. The differential regulation of the two beta-subunits by hCG could be mimicked by 3 isobutyl-1-methylxanthine, forskolin, and dibutyryl-cAMP, suggesting involvement of the intracellular cAMP pathway. Interestingly, H89 (a specific inhibitor of protein kinase A, PKA) could effectively block hCG- and forskolin-stimulated activin betaA expression at 10 micro M, but it was unable to reverse the inhibitory effects of hCG and forskolin on betaB expression. This suggests that the hCG-stimulated activin betaA expression is dependent on the activation of the cAMP-PKA pathway, whereas the inhibitory effect of hCG on activin betaB expression is likely mediated by PKA-independent pathway(s). PMID- 12538610 TI - Glucose intolerance and resistin expression in rat offspring exposed to ethanol in utero: modulation by postnatal high-fat diet. AB - High-fat diet and intrauterine growth retardation may predispose to obesity, insulin resistance, and type 2 diabetes. Because prenatal ethanol (ETOH) exposure causes intrauterine growth retardation, we investigated its interactions with postnatal high-fat diet on glucose tolerance and adipocyte-derived hormones in the rat offspring. High-fat-fed offspring had increased adiposity, serum leptin, and muscle uncoupling protein-3, but decreased adiponectin mRNA, compared with corresponding chow-fed groups. ETOH-exposed offspring had normal adiponectin, but increased resistin mRNA and protein, compared with controls, regardless of postnatal diet. Skeletal muscle glucose transporter-4 content was decreased after both ETOH exposure and high-fat feeding. Glycemic and insulin responses to an ip glucose challenge were equally increased in non-ETOH-exposed high-fat-fed offspring and in ETOH-exposed chow-fed offspring, with additive effects of ETOH and high-fat diet. Pancreatic insulin content was elevated only in non-ETOH exposed high-fat-fed offspring. The data suggest that high-fat diet worsens glucose intolerance in offspring of rats exposed to ETOH. Prenatal ETOH exposure and postnatal high-fat diet might cause insulin resistance through separate mechanisms, involving resistin and adiponectin, respectively. PMID- 12538612 TI - Two Inr elements are important for mediating the activity of the proximal promoter of the human gonadotropin-releasing hormone receptor gene. AB - Differential usage of several transcription start sites in the human GnRH receptor gene was evident in human brain and pituitary. To locate the promoter responsible for a cluster of the 3' CAP sites from -635 to -578 (relative to ATG) found in the pituitary, a proximal promoter element was identified at -677/-558 by 5' and 3' deletion mutant analysis. The promoter element drove a 13.1 +/- 0.6 fold increase in reporter gene activity in an orientation-dependent manner in the mouse gonadotrope-derived alphaT3-1 cells. Within the core promoter element, two functional AT-rich Inr motifs, interacting with the same protein factor with different affinities, were identified. By Southwestern blot analysis and competitive gel mobility shift assays, multiple nuclear factors (36-150 kDa) were found to interact specifically with the core promoter element. Interestingly, these nuclear proteins also interacted with a previously identified distal promoter of the human GnRH receptor gene. Taken together, our studies suggested that these two promoters share common protein factors to regulate transcription initiations at two different regions. Additional mechanisms are needed to modulate the efficiencies of individual promoters for developmental and/or tissue specific regulations. PMID- 12538611 TI - Sertoli and germ cell development in hypogonadal (hpg) mice expressing transgenic follicle-stimulating hormone alone or in combination with testosterone. AB - We recently created a novel transgenic (tg) model to examine the specific gonadal actions of FSH, distinct from LH effects, by expressing tg-FSH in gonadotropin deficient hypogonadal (hpg) mice. Using this unique in vivo paradigm, we now describe the postnatal cellular development in seminiferous tubules selectively stimulated by tg-FSH alone or combined with testosterone (T). In the alphabeta.6 line, tg-FSH stimulated the maturation and proliferation ( approximately 2-fold) of Sertoli cells in hpg testes. Total Sertoli cell numbers were also significantly increased (1.5-fold) independently of FSH effects by T treatment alone. Selective FSH activity in alphabeta.6 hpg testes increased total spermatogonia numbers 3-fold, which established a normal spermatogonia/Sertoli cell ratio. FSH also elevated meiotic spermatocyte numbers 7-fold, notably at pachytene (28-fold), but induced only limited numbers of postmeiotic haploid cells (absent in hpg controls) that arrested during spermatid elongation. In contrast, T treatment alone had little effect on postnatal spermatogonial proliferation but greatly enhanced meiotic progression with total spermatocytes increased 12-fold (pachytene 53-fold) relative to hpg testes, and total spermatid numbers 11-fold higher than tg-FSH hpg testes. Combining tg-FSH and T treatment had no further effect on Sertoli or spermatogonia numbers relative to FSH alone but had marked additive and synergistic effects on meiotic cells, particularly pachytene (107-fold more than hpg), to establish normal meiotic germ cell/Sertoli cell ratios. Furthermore, tg-FSH had a striking synergistic effect with T treatment on total spermatid numbers (19-fold higher than FSH alone), although spermatid to Sertoli cell ratios were not fully restored to normal, indicating elevated Sertoli cell numbers alone are insufficient to establish a maximal postmeiotic germ cell capacity. This unique model has allowed a detailed dissection of FSH in vivo activity alone or with T and provided compelling evidence that FSH effects on spermatogenesis are primarily via Sertoli and spermatogonial proliferation and the stimulation of meiotic and postmeiotic germ cell development in synergy with and dependent on T actions. PMID- 12538613 TI - The severe form of hypertension caused by the activating S810L mutation in the mineralocorticoid receptor is cortisone related. AB - A gain of function mutation resulting in the substitution of leucine for serine at codon 810 (S810L) in the human mineralocorticoid receptor (MR) is responsible for early-onset hypertension that is exacerbated in pregnancy. All steroids, including progesterone, that display antagonist properties when bound to the wild type MR are able to activate the mutant receptor (MR(L810)). These findings suggest that progesterone may contribute to the dramatic aggravation of hypertension in MR(L810) carriers during pregnancy. However, the steroid(s) responsible for hypertension in MR(L810) carriers (men and nonpregnant women) has not yet been identified. Here we show that cortisone and 11 dehydrocorticosterone, the main cortisol and corticosterone metabolites produced in the distal nephron, where sodium reabsorption stimulated by aldosterone takes place, bind with high affinity to MR(L810). The potency with which cortisone and 11-dehydrocorticosterone bind to the mutant MR contrasts sharply with their low wild-type MR-binding capacity. In addition, cotransfection assays demonstrate that cortisone and 11-dehydrocorticosterone are potent activators of the MR(L810) trans-activation function. Because the plasma concentration of cortisol in humans is about 30-fold higher than that of corticosterone, these findings strongly suggest that cortisone is one of the endogenous steroids responsible for early onset hypertension in men and nonpregnant women carrying the MR(L810) mutation. PMID- 12538614 TI - 12-lipoxygenase pathway increases aldosterone production, 3',5'-cyclic adenosine monophosphate response element-binding protein phosphorylation, and p38 mitogen activated protein kinase activation in H295R human adrenocortical cells. AB - Evidence suggests that the 12-lipoxygenase (LO) pathway mediates angiotensin II (Ang II)-induced aldosterone synthesis in adrenal glomerulosa cells. To study the mechanisms of 12-LO pathway on aldosterone synthesis, the human adrenocortical cell line, H295R, was transiently transfected with a mouse leukocyte type of 12 LO. Overexpression of 12-LO stimulated aldosterone production 2.7-fold as well as the reporter gene activity of CYP11B2 gene-encoding human aldosterone synthase by 5-fold over that in mock-transfected cells. Ang II further enhanced aldosterone production, which could be blocked by a 12-LO inhibitor, baicalein, in mock cells and cells overexpressing 12-LO. Ang II stimulated cAMP response element-binding protein (CREB) phosphorylation in a dose- and time-dependent fashion in parent H295R cells. Overexpression of 12-LO increased phosphorylation of CREB/activating transcription factor (ATF)-1 1.5-fold over that in mock cells under basal conditions. Ang II led to a further 5.2- and 7.5-fold increase in mock cells and 12-LO cells, respectively. Overexpression of 12-LO induced p38 MAPK activation. The 12-LO product, 12-hydroxyeicosatetraenoic acid, increased phosphorylation of CREB/ATF-1 3.6-fold and phosphorylation of p38 MAPK 8-fold over basal. The p38 MAPK inhibitor SB203580 inhibited Ang II- and 12-LO pathway-induced phosphorylated CREB/ATF-1, suggesting a role of p38 MAPK in Ang II and 12-LO pathway signaling. These results suggest that 12-LO stimulation leads to aldosterone production in H295R cells in part through activation of CREB/ATF-1 and p38 MAPK pathway. PMID- 12538615 TI - Agouti-related peptide, neuropeptide Y, and somatostatin-producing neurons are targets for ghrelin actions in the rat hypothalamus. AB - Ghrelin, the endogenous ligand of the GH secretagogue receptor, acts at central level to elicit GH release and regulate food intake. To elucidate the neural circuit that exerts its effects, we measured the expression of hypothalamic neuropeptides involved in weight regulation and GH secretion after ghrelin administration. Adult male rats, fed or fasted for 72 h, were treated centrally (intracerebroventicularly) with a single dose of ghrelin (5 micro g). After 2, 4, and 6 or 8 h, agouti-related peptide, melanin-concentrating hormone, neuropeptide Y, prepro-orexin, GHRH, and somatostatin mRNA levels were measured by in situ hybridization. We found that ghrelin increased agouti-related peptide and neuropeptide Y expression in the arcuate nucleus of the hypothalamus of fed and fasted rats. In contrast, no change was demonstrated in the mRNA levels of the other neuropeptides studied at any time evaluated. Finally, we examined the effect of ghrelin on GHRH and somatostatin mRNA levels in GH-deficient (dwarf) rats. Our results show that ghrelin increases somatostatin mRNA levels in the hypothalamus of these rats. This study furthers our understanding of the molecular basis and mechanisms involved in the effect of ghrelin on food intake and GH secretion. PMID- 12538616 TI - Dronerarone acts as a selective inhibitor of 3,5,3'-triiodothyronine binding to thyroid hormone receptor-alpha1: in vitro and in vivo evidence. AB - Dronedarone (Dron), without iodine, was developed as an alternative to the iodine containing antiarrhythmic drug amiodarone (AM). AM acts, via its major metabolite desethylamiodarone, in vitro and in vivo as a thyroid hormone receptor alpha(1) (TRalpha(1)) and TRbeta(1) antagonist. Here we investigate whether Dron and/or its metabolite debutyldronedarone inhibit T(3) binding to TRalpha(1) and TRbeta(1) in vitro and whether dronedarone behaves similarly to amiodarone in vivo. In vitro, Dron had a inhibitory effect of 14% on the binding of T(3) to TRalpha(1), but not on TRbeta(1). Desethylamiodarone inhibited T(3) binding to TRalpha(1) and TRbeta(1) equally. Debutyldronedarone inhibited T(3) binding to TRalpha(1) by 77%, but to TRbeta(1) by only 25%. In vivo, AM increased plasma TSH and rT(3), and decreased T(3). Dron decreased T(4) and T(3), rT(3) did not change, and TSH fell slightly. Plasma total cholesterol was increased by AM, but remained unchanged in Dron-treated animals. TRbeta(1)-dependent liver low density lipoprotein receptor protein and type 1 deiodinase activities decreased in AM treated, but not in Dron-treated, animals. TRalpha(1)-mediated lengthening of the QTc interval was present in both AM- and Dron-treated animals. The in vitro and in vivo findings suggest that dronedarone via its metabolite debutyldronedarone acts as a TRalpha(1)-selective inhibitor. PMID- 12538617 TI - Environmental salinity regulates receptor expression, cellular effects, and circulating levels of two antagonizing hormones, 1,25-dihydroxyvitamin D3 and 24,25-dihydroxyvitamin D3, in rainbow trout. AB - In freshwater-adapted rainbow trout, intestinal cells (enterocytes) possess receptors for 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] in the basolateral membrane, and respond to treatment with 1,25(OH)(2)D(3) with increased intracellular calcium concentrations. No receptors are found for the antagonizing hormone 24,25-dihydroxyvitamin D(3) [24,25(OH)(2)D(3)] at the enterocyte basolateral membrane, and it has no effect on enterocyte calcium homeostasis. After acclimation to seawater, however, the enterocyte membrane receptors for 1,25(OH)(2)D(3) are down-regulated and specific binding for 24,25(OH)(2)D(3) appears, which is further up-regulated with time spent in seawater. This shift in receptor expression is concurrent with an increased sensitivity of the enterocytes to 24,25(OH)(2)D(3) and a decreased sensitivity to 1,25(OH)(2)D(3). This results in a partial inhibition of intracellular calcium uptake, which would be beneficial when inhabiting a calcium-rich environment like seawater. PMID- 12538618 TI - Identification of a truncated dual oxidase 2 (DUOX2) messenger ribonucleic acid (mRNA) in two rat thyroid cell lines. Insulin and forskolin regulation of DUOX2 mRNA levels in FRTL-5 cells and porcine thyrocytes. AB - The Duox2 flavoprotein is strongly expressed in the thyroid gland, where it plays a critical role in the synthesis of thyroid hormones likely by providing thyroperoxidase with H(2)O(2). A truncated DUOX2 mRNA was isolated from the rat thyroid cell line FRTL-5. The cDNA sequence predicted an open reading frame of 1458 bp, encoding a polypeptide of 486 amino acids corresponding to the carboxyl fragment of the Duox2 flavoprotein. The truncated form of DUOX2 mRNA, expressed in another rat thyroid cell line, the PC Cl3 cell line, was absent from Fischer rat thyroid glands. Although it was expressed in both cell lines to a greater extent than normal mRNA, it failed to support protein synthesis in an in vitro translation system. Insulin increased the levels of both normal and truncated DUOX2 mRNA in FRTL-5 cells grown in TSH-free medium containing a low concentration of serum. The stimulating effect of insulin on DUOX2 mRNA expression was reproduced in pig thyroid follicles in primary culture. The presence of insulin in the culture medium converted forskolin from a stimulator to an inhibitor in FRTL-5 cells maintained in low serum conditions, but not in porcine thyrocytes in primary culture. PMID- 12538619 TI - 5alpha-androstane-3alpha,17beta-diol is formed in tammar wallaby pouch young testes by a pathway involving 5alpha-pregnane-3alpha,17alpha-diol-20-one as a key intermediate. AB - The synthetic pathway by which 5alpha-androstane-3alpha,17beta-diol (5alpha adiol) is formed in the testes of tammar wallaby pouch young was investigated by incubating testes from d 20-40 males with various radioactive precursors and analyzing the metabolites by thin-layer chromatography and HPLC. [(3)H]Progesterone was converted to 17-hydroxyprogesterone, which was converted to 5alpha-adiol by two pathways: One involves the formation of testosterone and dihydrotestosterone as intermediates, and the other involves formation of 5alpha pregnane-3alpha,17alpha-diol-20-one (5alpha-pdiol) and androsterone as intermediates. Formation of 5alpha-adiol from both [(3)H]testosterone and [(3)H]progesterone was blocked by the 5alpha-reductase inhibitor 4MA. The addition of nonradioactive 5alpha-pdiol blocked the conversion of [(3)H]progesterone to 5alpha-adiol, and [(3)H]5alpha-pdiol was efficiently converted to androsterone and 5alpha-adiol. We conclude that expression of steroid 5alpha-reductase in the developing wallaby testes allows formation of 5alpha-reduced androgens by a pathway that does not involve testosterone as an intermediate. PMID- 12538620 TI - Essential role of insulin-like growth factor I receptor in insulin-induced fetal brown adipocyte differentiation. AB - To define the specific role of IGF-I receptor (IGF-IR) in adipogenic and thermogenic differentiation of brown adipocytes during late fetal life, we have established immortalized brown adipocyte cell lines from fetuses of IGF-IR deficient mice (IGF-IR(-/-)) as well as from wild-type mice (IGF-IR(+/+)). IGF IR(-/-) cells showed an increased insulin sensitivity regarding insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation despite a substantial reduction in IRS-1 protein content. Furthermore, insulin-induced total and IRS-1-associated phosphatidylinositol 3-kinase activities were augmented in IGF-IR-deficient cells compared with wild-type cells. Downstream phosphatidylinositol 3-kinase activation of Akt, but not p70s6 kinase, were elicited at lower doses of insulin in IGF-IR(-/-) brown adipocytes. Activation of protein kinase Czeta by insulin was similar in both cell types as was insulin-induced glucose uptake. Treatment of wild-type brown adipocytes with insulin for 12 h up-regulated fatty acid synthase (FAS) and adipocyte determination and differentiation (ADD1/SREBP) mRNAs; this effect was impaired in the absence of IGF-IR. At the protein level, insulin increased FAS content and the amount of the mature form of adipocyte determination and differentiation (ADD1/SREBP) in the nucleus in wild-type cells, but not in IGF-IR(-/-) cells. Furthermore, 24 h of insulin stimulation induced the expression of both uncoupling protein-1 and CCAAT/enhancer-binding protein alpha (C/EBPalpha) in wild-type brown adipocytes; these effects were abolished in IGF-I-R(-/-) cells. Retrovirus-mediated reexpression of peroxisomal proliferator activated receptor gamma (PPARgamma) in IGF-IR(-/-) brown adipocytes could overcome FAS mRNA impairment, bypassing insulin signaling. However, insulin further increased FAS mRNA expression in C/EBPalpha-IGF-IR(-/-) cells, but not in PPARgamma-IGF-IR(-/-) cells. In addition, fetal brown adipocytes lacking IGF-IR up-regulated uncoupling protein-1 expression in the absence of insulin when PPARgamma, but not C/EBPalpha, was overexpressed. These data provide strong evidence for a critical role of IGF-IR in the differentiation of the brown adipocyte phenotype in fetal life; this effect is mimicked by PPARgamma in an insulin-independent manner. PMID- 12538621 TI - Importance of the paraventricular nucleus of the hypothalamus as a component of a neural pathway between the brain and the testes that modulates testosterone secretion independently of the pituitary. AB - We previously reported that in adult male rats, the intracerebroventricular (icv) injection of corticotropin-releasing factor (CRF) or the beta-adrenergic agonist isoproterenol (ISO) significantly inhibited the ability of human chorionic gonadotropin (hCG) to stimulate testosterone (T) secretion. The finding that this phenomenon also took place when LH release had been blocked with an LHRH antagonist suggested that icv CRF and ISO did not alter Leydig cell function by influencing the activity of pituitary gonadotrophs. We therefore proposed the existence of a neural pathway connecting the brain to the testes, whose activation by icv CRF or ISO interfered with T secretion. Based on the intratesticular injection of the transganglionic tracer pseudorabies virus, we recently identified the paraventricular nucleus (PVN) of the hypothalamus as a component of this neural link. The aim of the present work was to investigate the functional role of this brain area in mediating the ability of CRF and ISO to inhibit the ability of hCG to stimulate T secretion. We first demonstrated that local microinfusion of CRF or ISO directly into the PVN mimicked the effect of their icv injection, suggesting that the PVN does indeed represent a site of action of ISO and CRF in altering Leydig cell responsiveness to gonadotropin. In contrast, neither CRF nor ISO microinfusion into the central amygdala or the frontal cortex influenced hCG-stimulated T secretion. To further investigate the role of the PVN in ISO- and CRF-induced blunting of hCG stimulation of T, we determined the effect of icv CRF or ISO on testicular activity of rats with electrolytic lesions of the PVN. These lesions, which did not in themselves influence Leydig cell responsiveness to hCG, blocked the effect of both icv ISO and CRF on hCG-induced T release. Collectively, these results support the hypothesis that CRF- and ISO-induced activation of cells in the area of the PVN decreases the ability of gonadotropin to release T and suggests that this nucleus represents an important site of the proposed neural connection between the brain and the testes. PMID- 12538622 TI - Biological activity of 17beta-estradiol-3-sulfate in ovine fetal plasma and uptake in fetal brain. AB - In sheep, the fetal hypothalamus-pituitary-adrenal axis plays a central role in the initiation of parturition. We have reported that estradiol dramatically increases the activity of the fetal hypothalamus-pituitary-adrenal (HPA) axis. Sulfoconjugated estrogens are known to circulate in high concentrations in fetal plasma. We have reported the expression and abundant activity of steroid sulfatase within the fetal brain regions important for HPA axis control, and we have proposed that sulfoconjugated estrogens in fetal plasma are deconjugated (and therefore converted to a biologically active form) in fetal brain. The present study was designed to test the hypothesis that exogenous estradiol-3 sulfate stimulates HPA axis activity in late gestation fetal sheep and that it is concentrated by fetal brain tissue. We infused estradiol-3-sulfate iv into fetal sheep (125-135 d gestation; term = 147 d) at rates of 0, 0.25, and 1.0 mg/d for 5 d and performed serial sampling of fetal blood before and at the end of the infusion periods. Infusions increased fetal plasma estradiol-3-sulfate concentrations and produced dose-related increases in HPA axis activity. The action of the steroid on the fetal brain was also demonstrated as dose-related increases in the abundance of Fos in fetal cerebellum. In a second study we measured the uptake of sulfoconjugated and unconjugated estrogen (estrone 3sulfate and estrone, respectively) into the fetal brain (124-128 d gestation) in vivo. Both forms of estrogen were concentrated in fetal brain, with the uptake of estrone greater than that of estrone-3-sulfate. We conclude that sulfoconjugated estrogens augment fetal HPA axis activity and that they can cross the fetal blood brain barrier. We propose that in late gestation the large circulating pool of sulfoconjugated estrogen is a biologically important source of active hormone that might play a role in the timing of parturition in sheep. PMID- 12538623 TI - Hepatocyte retinoid X receptor-alpha-deficient mice have reduced food intake, increased body weight, and improved glucose tolerance. AB - Hepatocyte retinoid X receptor (RXR)alpha-deficient mice and wild-type mice were fed either a regular or a high-saturated-fat diet for 12 wk to study the functional role of hepatocyte RXRalpha in fatty acid and carbohydrate metabolism. Food intake was significantly reduced in hepatocyte RXRalpha-deficient mice when either diet was used. The amount of food intake was negatively associated with serum leptin level. Although mutant mice ate less, body weight and fat content were significantly higher in mutant than wild-type mice. Examination of the expression of peroxisome proliferator-activated receptor-alpha target genes indicated that the peroxisome proliferator-activated receptor-alpha-mediated pathway was compromised in the mutant mice, which, in turn, might affect fatty acid metabolism and result in increased body weight and fat content. Although mutant mice were obese, they demonstrated the same degree of insulin sensitivity and the same level of serum insulin as the wild-type mice. However, these mutant mice have improved glucose tolerance. To explore a mechanism that may be responsible for the improved glucose tolerance, serum IGF-I level was examined. Serum IGF-1 level was significantly increased in mutant mice compared with wild type mice. Taken together, hepatocyte RXRalpha deficiency increases leptin level and reduces food intake. Those mice also develop obesity, with an unexpected improvement of glucose tolerance. The result also suggests that an increase in serum IGF-I level might be one of the mechanisms leading to improved glucose tolerance in hepatocyte RXRalpha-deficient mice. PMID- 12538624 TI - Extracellular signal-regulated kinase and c-Src, but not Jun N-terminal kinase, are involved in basal and gonadotropin-releasing hormone-stimulated activity of the glycoprotein hormone alpha-subunit promoter. AB - Addition of a GnRH agonist (GnRH-A) to alphaT3-1 cells stimulates different MAPK cascades: ERK, Jun N-terminal kinase (JNK), and p38. Activation of JNK, ERK, and p38 shows a unique fold activation ratio of 25:12:2, which might encode signal specificity. ERK is translocated to the nucleus within 20 min with a peak at 120 min of GnRH-A stimulation. We used the human alpha-subunit promoter linked to chloramphenicol acetyl transferase (alphaCAT) to examine the role of ERK, JNK, and c-Src, which is implicated in MAPK activation, in basal and GnRH-stimulated alphaCAT. Addition of GnRH-A resulted in a 3-fold increase in alphaCAT, whereas the Ca(2+) ionophore ionomycin and the protein kinase C (PKC) activator 12-O tetradecanoylphorbol-13-acetate (TPA) had no effect. Addition of GnRH-A and TPA, but not GnRH-A and ionomycin, produced a synergistic response, whereas removal of Ca(2+), but not down-regulation of TPA-sensitive PKCs, abolished GnRH-A stimulated alphaCAT. Thus, regulation of alpha-promoter activity by GnRH is Ca(2+) dependent and is further augmented by PKC. Cotransfection of alphaCAT and constitutively active or dominant negative plasmids of ERK and JNK cascade members, or the use of the ERK inhibitor PD98059, revealed that ERK, but not JNK, is involved in basal and GnRH-A-stimulated alphaCAT. Because c-Src participates in MAPK activation by GnRH, we also studied its role. Cotransfection of alphaCAT and the dominant negative form of c-Src or incubation with the c-Src inhibitor PP1 reduced GnRH-A-stimulated alphaCAT. The 5'-deletion analysis revealed that the -846/-420 region participated in basal alpha-transcription. In addition, the 346/-156 region containing the pituitary glycoprotein hormone basal element, alpha-basal elements, glycoprotein-specific element, and upstream response element is involved in basal and GnRH-A-stimulated alphaCAT. ERK contribution to GnRH maps to -346/-280 containing the pituitary glycoprotein hormone basal element and alpha-basal elements 1/2. Surprisingly, although c-Src is involved in GnRH-A-stimulated ERK, its involvement is mapped to another region (-280/-180) containing the glycoprotein-specific element. Thus, ERK and c-Src but not JNK are involved in basal and GnRH-A-stimulated-alphaCAT, whereas c-Src contribution is independent of ERK activation. PMID- 12538625 TI - Resistance of human ovarian cancer cells to tumor necrosis factor alpha is a consequence of nuclear factor kappaB-mediated induction of Fas-associated death domain-like interleukin-1beta-converting enzyme-like inhibitory protein. AB - The purpose of the present studies was to examine the role and regulation of Fas associated death domain-like IL-1- converting enzyme-like inhibitory protein [FLIP; long (FLIP(L)) and short (FLIP(S)) forms] in human ovarian epithelial cancer cells by TNFalpha and their significance in the resistance of the cells to the proapoptotic action of the cytokine. OV2008, A2780-s, and OVCAR-3 cells were cultured in serum-free media with or without cycloheximide (CHX, 10 micro g/ml) +/- TNFalpha (5, 10, 20 ng/ml) or transfected with a mammalian expression vector containing either a dominant negative inhibitor kappaB (IkappaB), FLIP(S) sense or antisense cDNA and cultured with or without TNFalpha. In the presence of CHX, TNFalpha increased caspase-8 and -3 cleavage and apoptosis. It also induced IkappaB phosphorylation, nuclear factor kappaB activation, and the expression of FLIP(S) but not of FLIP(L). Overexpression of dominant negative IkappaB attenuated TNFalpha-induced FLIP(S) expression and enhanced TNFalpha-induced apoptosis. Apoptosis induced by TNFalpha and CHX was facilitated by FLIP(S) antisense expression but attenuated by sense transfection. This study demonstrates that TNFalpha up-regulates FLIP(S) expression, and this effect is mediated by the activation of nuclear factor kappaB. The induction of FLIP(S) expression by TNFalpha might contribute to the resistance of ovarian epithelial cancer cells to the proapoptotic action of the cytokine. PMID- 12538626 TI - Deletion of V335 from the L2 domain of the insulin receptor results in a conformationally abnormal receptor that is unable to bind insulin and causes Donohue's syndrome in a human subject. AB - An infant with Donohue's syndrome (leprechaunism) was found to be homozygous for an in-frame trinucleotide deletion within the insulin receptor gene resulting in the deletion of valine 335. When transiently transfected into Chinese hamster ovary cells, mutant receptor was produced in a mature form, but at significantly lower levels compared with wild-type receptor. Cell surface biotinylation experiments revealed that significant amounts of the DeltaV335 receptor were expressed on the cell surface. Despite this, cells expressing this receptor showed no significant insulin binding or ligand-induced receptor autophosphorylation. Although the DeltaV335 receptor was capable of being immunoprecipitated with antibodies directed against the beta-subunit of the receptor, the mutant receptor could not be recognized by a panel of antibodies directed against different epitopes of the alpha-subunit, suggesting that the loss of V335 results in a major conformational alteration in the receptor alpha subunit. This would be predicted by the positioning of V335 at a critical location within a strand that provides the main rigid scaffold for the two beta sheet faces of the L2 domain of the receptor. The severe biochemical and clinical consequences of this novel mutation, which occur despite substantial expression on the cell surface, emphasize the crucial role of the L2 domain in ligand binding by the insulin receptor. PMID- 12538627 TI - Novel signal transduction pathway for luteinizing hormone and its interaction with insulin: activation of Janus kinase/signal transducer and activator of transcription and phosphoinositol 3-kinase/Akt pathways. AB - The actions of LH are mediated through a single class of cell surface LH/human chorionic gonadotropin receptor, which is a member of the G protein-coupled receptor family. In the present study we showed that LH induced rapid tyrosine phosphorylation and activation of the Janus kinase 2 (JAK2) in rat ovary. Upon JAK2 activation, tyrosine phosphorylation of signal transducer and activator of transcription-1 (STAT-1), STAT-5b, insulin receptor substrate-1 (IRS-1), and Src homology and collagen homology (Shc) were detected. In addition, LH induced IRS 1/phosphoinositol 3-kinase and Shc /growth factor receptor-binding protein 2 (Grb2) associations and downstream AKT (protein kinase B, homologous to v-AKT) serine phosphorylation and ERK tyrosine phosphorylation, respectively. The simultaneous infusion of insulin and LH induced higher phosphorylation levels of JAK2, STAT5b, IRS-1, and AKT compared with each hormone alone in the whole ovary of normal rats. By immunohistochemistry we demonstrated that these late events take place in follicular cells and both external and internal theca. These results indicate a new signal transduction pathway for LH and show that there is positive cross-talk between the insulin and LH signaling pathways at the level of phosphoinositol 3-kinase/AKT pathway in this tissue. PMID- 12538628 TI - Long-term apoptotic cell death process with increased expression and activation of caspase-3 and -6 in adult rat germ cells exposed in utero to flutamide. AB - Although it is established that in utero exposure to antiandrogenic compounds such as flutamide induces hypospermatogenesis in adult male rat offspring, the cellular and molecular mechanisms remain to be investigated. By using adult rats exposed in utero to flutamide (0.4, 2, 10 mg/kg.d) as a model, we show that the hypospermatogenesis could be related to a chronic apoptotic cell death process associated with a long-term increase in caspase-3 and -6 expression and activation in germ cells. The number of apoptotic (terminal deoxynucleotidyl transferase-mediated deoxyuridine positive) adult germ cells was dependent on the dose of flutamide. The apoptotic germ cell death process could be related to an increased expression and activation of effector caspases-3 and -6. Procaspases-3 and -6 were immunodetected in germ cells from both untreated or flutamide-treated rats, whereas cleaved active caspase-3 was detected exclusively in germ cells from adult rat exposed in utero to flutamide. Exposure to the antiandrogen increased in a dose-dependent manner as caspase-3 and -6 mRNA (in RT-PCR approaches) as well as procaspase-3 and -6 protein (in Western blotting analyses) levels in the adult rat testis. Flutamide also activates procaspases. Indeed, whereas cleaved active caspase-3 and -6 proteins were absent in control animals, they were detected in adult rat testes exposed in utero to flutamide. Our results show that whereas the apoptotic germ cell death process associated with the increased caspase expression and activation in adult rat germ cells was chronic and nonreversible when exposure to flutamide occurred in utero, it was transient when such an exposure occurred during adulthood. Indeed, although an increase in caspase-3 and -6 mRNA and procaspase-3 and -6 protein levels was observed in germ cells after 3 d of exposure to flutamide, 1-2 wk after the cessation of the antiandrogen exposure, the caspase mRNA and procaspase protein levels were back to control. Active cleaved caspase-3 and -6 protein appeared following the exposure to the antiandrogen, whereas they disappeared at cessation of exposure to flutamide. In summary, the present findings indicate that in utero exposure to the antiandrogen induced in the adult rat testes a chronic apoptotic germ cell death associated with a long-term increase in the expression and activation in germ cells of caspases-3 and -6, two key components in the death machinery. PMID- 12538629 TI - Distribution of vesicular glutamate transporter-2 messenger ribonucleic Acid and protein in the septum-hypothalamus of the rat. AB - The excitatory neurotransmitter glutamate is involved in the control of most, perhaps all, neuroendocrine systems, yet the sites of glutamatergic neurons and their processes are unknown. Here, we used in situ hybridization and immunohistochemistry for the neuron-specific vesicular glutamate transporter-2 (VGLUT2) to identify the neurons in female rats that synthesize the neurotransmitter glutamate as well as their projections throughout the septum hypothalamus. The results show that glutamatergic neurons are present in the septum-diagonal band complex and throughout the hypothalamus. The preoptic area and ventromedial and dorsomedial nuclei are particularly rich in glutamatergic neurons, followed by the supraoptic, paraventricular, and arcuate nuclei, whereas the suprachiasmatic nucleus does not express detectable amounts of VGLUT2 mRNA. Immunoreactive neurites are seen in very high densities in all regions analyzed, particularly in the preoptic region, followed by the ventromedial, dorsomedial, and arcuate nuclei as well as the external layer of the median eminence, whereas the mammillary complex does not exhibit VGLUT2 immunoreactivity. Many VGLUT2 immunoreactive fibers also contained synaptophysin, suggesting that the transporter is indeed localized to presynaptic terminals. Together, the results identify glutamatergic cell bodies throughout the septum-hypothalamus in region specific patterns and show that glutamatergic nerve terminals are present in very large numbers such that most neurons in these brain regions can receive glutamatergic input. We examined the GnRH system as an example of a typical neuroendocrine system and could show that the GnRH perikarya are closely apposed by many VGLUT2-immunoreactive boutons, some of which also contained synaptophysin. The presence of VGLUT2 mRNA-containing cells in specific nuclei of the hypothalamus indicates that many neuroendocrine neurons coexpress glutamate as neurotransmitter, in addition to neuropeptides. These systems include the oxytocin, vasopressin, or CRH neurons as well as many others in the periventricular and mediobasal hypothalamus. The presence of VGLUT2 mRNA in steroid-sensitive regions of the hypothalamus, such as the anteroventral periventricular, paraventricular, or ventromedial nuclei indicates that gonadal and adrenal steroid can directly alter the functions of these glutamatergic neurons. PMID- 12538630 TI - Induction of thyroid-stimulating hormone receptor autoimmunity in hamsters. AB - Female Chinese hamsters (n = 10) were immunized with Chinese hamster ovary (CHO) cells that expressed the human TSH receptor (TSHR) to generate a model of Graves' disease. TSHR-autoantibodies (TSHR-Ab) were determined by CHO-TSHR. Two hamsters with stimulating TSHR-Ab showed thyrocyte hypertrophy associated with a focal lymphocytic infiltration. CHO-TSHR were then stimulated with interferon gamma to enhance major histocompatibility complex class II expression. However, after immunization no stimulating TSHR-Ab were detected, but blocking TSHR-Ab were found in three of five animals. The thyroid glands from these hamsters showed marked thinning of thyroid epithelial cells, indicative of early thyroid atrophy consistent with a TSHR blocking antibody, but no lymphocytic infiltration. Lastly, female Armenian hamsters were immunized with an adenovirus construct incorporating wild-type TSHR. High titers of TSHR-Ab were induced effectively, but the thyroid hypertrophy observed was not associated with a lymphocyte infiltration. In summary, we demonstrated that the hamster could serve as a model of TSHR autoimmunity and that an adenoviral vector produced higher levels of TSHR Ab than more conventional immunization with cells. The data also indicated that the intrathyroidal cellular immunity in this model was not related to TSHR-Ab formation and was an independent reflection of the T-cell immune response to TSHR antigen. PMID- 12538631 TI - Statins augment vascular endothelial growth factor expression in osteoblastic cells via inhibition of protein prenylation. AB - Statins such as simvastatin are 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors that inhibit cholesterol synthesis. We presently investigated statin effects on vascular endothelial growth factor (VEGF) expression in osteoblastic cells. Hydrophobic statins including simvastatin, atorvastatin, and cerivastatin but not a hydrophilic statin, pravastatin-markedly increased VEGF mRNA abundance in nontransformed osteoblastic cells (MC3T3-E1). Simvastatin (10(-6) M) time dependently augmented VEGF mRNA expression in MC3T3-E1 cells, mouse stromal cells (ST2), and rat osteosarcoma cells (UMR-106). According to heterogeneous nuclear RNA and Northern analyses, 10(-6) M simvastatin stimulated gene expression for VEGF in MC3T3-E1 cells without altering mRNA stability. Transcriptional activation of a VEGF promoter-luciferase construct (-1128 to +827), significantly increased by simvastatin administration. As demonstrated by gel mobility shift assay, simvastatin markedly enhanced the binding of hypoxia-responsive element protein complexes. These results indicate that the stimulation of the VEGF gene by simvastatin in MC3T3-E1 cells is transcriptional in nature. VEGF secretion into medium was increased in MC3T3-E1 by 10(-6) M simvastatin. Pretreating MC3T3 E1 cells with mevalonate or geranylgeranyl pyrophosphate, a mevalonate metabolite, abolished simvastatin-induced VEGF mRNA expression; manumycin A, a protein prenylation inhibitor, mimicked statin effects on VEGF expression. The effect of simvastatin was blocked by pretreatment with wortmannin and LY294002, specific phosphatidylinositide-3 kinase inhibitors. Simvastatin enhanced mineralized nodule formation in culture, whereas coincubation with mevalonate, geranylgeranyl pyrophosphate, LY294002, or VEGF receptor 2 inhibitor (SU1498) abrogated statin-induced mineralization. Thus, statins stimulate VEGF expression in osteoblasts via reduced protein prenylation and the phosphatidylinositide-3 kinase pathway, promoting osteoblastic differentiation. PMID- 12538632 TI - Reduced expression of the cytokine transducer gp130 inhibits hormone secretion, cell growth, and tumor development of pituitary lactosomatotrophic GH3 cells. AB - Two of the most potent cytokines that regulate anterior pituitary cell function are leukemia inhibitory factor and IL-6. These and others like IL-11 and ciliary neurotrophic factor are referred to as the gp130 cytokines because they share the gp130 glycoprotein as a common receptor initial signal transducer. We and others have shown that gp130 cytokines and their receptors are expressed and functional in normal and tumoral anterior pituitary cells. To study the role of gp130 cytokines in tumorigenic process, we generated gp130 cDNA gp130 sense and gp130 antisense (gp130-AS) transfected stable clones derived from lactosomatotroph GH3 cells. We examined hormone secretion and cell proliferation of these clones as well as their tumorigenic properties in athymic nude mice. Although gp130-AS clones, which have low gp130 levels and impaired signal transducer and activator of transcription 3 activity and suppressor of cytokine signaling-3 expression, showed reduced proliferation and hormone secretion (GH and prolactin) in response to gp130 cytokines, they had a normal response to gp130-independent stimuli. Moreover, gp130-AS clones showed a severely impaired in vivo tumor development. In contrast, the overexpressing gp130 clones (gp130 sense) showed no differences, compared with cells transfected with control vector. Thus, the present study provides new evidence supporting a link between gp130 and pituitary abnormal growth. PMID- 12538633 TI - Global transcription profiling of estrogen activity: estrogen receptor alpha regulates gene expression in the kidney. AB - Estrogen receptors (ERs) are expressed in numerous organs, although only a few organs are considered classical targets for estrogens. We have completed a systematic survey of estrogen regulation of approximately 10,000 genes in 13 tissues from wild-type and ERbetaKO mice treated sc with vehicle or 17beta estradiol (E2) for 6 wk. The uterus and pituitary had the greatest number of genes regulated by E2, whereas the kidney had the third largest number of regulated genes. In situ hybridizations localized E2 regulation in the kidney to the juxtamedullary region of the cortex in both the mouse and rat. The ED(50) for gene inductions in the kidney was 3 micro g/kg.d, comparable with the 2.4 micro g/kg.d ED(50) for c-fos induction in the uterus. E2 regulations in the kidney were intact in ERbetaKO mice, and the ERalpha-selective agonist propylpyrazole triol acted similarly to E2, together suggesting an ERalpha-mediated mechanism. Several genes were induced within 2 h of E2 treatment, suggesting a direct activity of ERalpha within the kidney. Finally, the combination of the activation function (AF)1-selective agonist tamoxifen plus ERalphaKO(CH) mice expressing an AF1-deleted version of ERalpha allowed delineation of genes with differing requirements for AF1 or AF2 activity in the kidney. PMID- 12538634 TI - Thyroid hormone regulates hepatic triglyceride mobilization and apolipoprotein B messenger ribonucleic Acid editing in a murine model of congenital hypothyroidism. AB - Thyroid hormone modulates the expression of numerous genes that in turn regulate lipoprotein metabolism in vivo. We have examined the thyroid hormone-dependent regulation of apolipoprotein B (apoB) RNA editing in a strain of congenitally hypothyroid mice (Pax8(-/-)) that lacks thyroid follicular cells. Neonatal Pax8( /-) mice demonstrate an approximately 10-fold increase in hepatic triglyceride content associated with a decrease in hepatic apoB RNA editing. Thyroid hormone administration resulted in hepatic triglyceride mobilization in conjunction with an increase in hepatic, but not intestinal, apoB RNA editing and without changing total apoB RNA abundance. ApoB RNA editing is mediated by a multicomponent enzyme complex whose catalytic core contains two proteins, apobec-1 and apobec-1 complementation factor (ACF). Hepatic ACF mRNA and protein abundance decreased in Pax8(-/-) mice, with restoration after thyroid hormone administration, whereas apobec-1 mRNA and protein abundance were unchanged. Immunohistochemical analysis revealed increased staining intensity of ACF within hepatocyte nuclei of treated mice, findings confirmed by Western analysis of isolated nuclei. In vitro RNA editing assays demonstrated that supplementation with recombinant ACF alone restored enzymatic activity of S100 extracts from hypothyroid, Pax8(-/-) mice. These data demonstrate that thyroid hormone modulates murine hepatic lipoprotein metabolism in association with tissue-specific effects on apoB RNA editing mediated through alterations in ACF gene expression. PMID- 12538635 TI - Autosomal dominant growth hormone deficiency disrupts secretory vesicles in vitro and in vivo in transgenic mice. AB - Autosomal dominant GH deficiency type II (IGHDII) is often associated with mutations in the human GH gene (GH1) that give rise to products lacking exon-3 ((Deltaexon3)hGH). In the heterozygous state, these act as dominant negative mutations that prevent the release of human pituitary GH (hGH). To determine the mechanisms of these dominant negative effects, we used a combination of transgenic and morphological approaches in both in vitro and in vivo models. Rat GC cell lines were generated expressing either wild-type GH1 (WT-hGH-GC) or a genomic GH1 sequence containing a G->A transition at the donor splice site of IVS3 ((Deltaexon3)hGH-GC). WT-hGH-GC cells grew normally and produced equivalent amounts of human and rGH packaged in dense-cored secretory vesicles (SVs). In contrast, (Deltaexon3)hGH-GC cells showed few SVs but accumulated secretory product in amorphous cytoplasmic aggregates. They produced much less rGH and grew more slowly than WT-hGH-GC cells. When cotransfected with an enhanced green fluorescent protein construct (GH-eGFP), which copackages with GH in SVs, WT-hGH GC cells showed normal electron microscopy morphology and SV movements, tracked with total internal reflectance fluorescence microscopy. In contrast, coexpression of (Deltaexon3)hGH with GH-eGFP abolished the vesicular targeting of GH-eGFP, which instead accumulated in static aggregates. Transgenic mice expressing (Deltaexon3)hGH in somatotrophs showed an IGHD-II phenotype with mild to severe pituitary hypoplasia and dwarfism, evident at weaning in the most severely affected lines. Hypothalamic GHRH expression was up-regulated and somatostatin expression reduced in (Deltaexon3)hGH transgenic mice, consistent with their profound GHD. Few SVs were detectable in the residual pituitary somatotrophs in (Deltaexon3)hGH transgenic mice, and these cells showed grossly abnormal morphology. A low copy number transgenic line showed a mild effect relatively specific for GH, whereas two severely affected lines with higher transgene copy numbers showed early onset, widespread pituitary damage, macrophage invasion, and multiple hormone deficiencies. These new in vitro and in vivo models shed new light on the cellular mechanisms involved in IGHDII, as well as its phenotypic consequences in vivo. PMID- 12538636 TI - Rat anterior pituitary folliculostellate cells are targets of interleukin-1beta and a major source of intrapituitary follistatin. AB - Folliculostellate cells of the anterior pituitary are postulated to be an important source of factors, such as follistatin, that regulate pituitary function by intercellular communication. To gain further insight into the function of this cell type, folliculostellate cells were enriched from cultured rat anterior pituitary cells, and an immortalized cell line designated FS/D1h was established and characterized. These FS/D1h cells express S100 immunoreactivity and produce IL-6 but not pituitary hormones such as GH, ACTH, FSH, and LH. Importantly, FS/D1h cells express large amounts of follistatin mRNA and secrete the protein, as quantified indirectly by the amount of [(125)I]activin A immunoprecipitated with a follistatin antiserum. The FS/D1h cells also express alpha, betaA, and betaB inhibin/activin subunit mRNAs, but whether they produce the corresponding activins and inhibins has not been determined. The response of FS/D1h cells to agents thought to modulate folliculostellate cell function was evaluated. IL-1beta (0.005-5 nM) stimulated the secretion of follistatin and increased mRNA expression. In parallel, IL-6 secretion was stimulated. Dexamethasone, pituitary adenylate cyclase-activating polypeptide(1-27), and lipopolysaccharide but not testosterone, 12-O-tetradecanoylphorbol-13-acetate, or forskolin also increased follistatin secretion. Surprisingly, activin had no effect on follistatin mRNA levels, despite the fact that FS/D1h cells express ActRII, ActRIIB, and ALK-4 (ActRIB). Activin, on the other hand, induced Smad7 mRNA accumulation and exerted an antiproliferative effect on FS/D1h cells. Altogether, these observations support the possibility that follistatin originating from folliculostellate cells participates in mediating the effects of IL-1beta, glucocorticoids, and other agents on the response of pituitary cells to activins. PMID- 12538637 TI - Endocytosing the death sentence. PMID- 12538638 TI - Traffic-independent function of the Sar1p/COPII machinery in proteasomal sorting of the cystic fibrosis transmembrane conductance regulator. AB - Newly synthesized proteins that do not fold correctly in the ER are targeted for ER-associated protein degradation (ERAD) through distinct sorting mechanisms; soluble ERAD substrates require ER-Golgi transport and retrieval for degradation, whereas transmembrane ERAD substrates are retained in the ER. Retained transmembrane proteins are often sequestered into specialized ER subdomains, but the relevance of such sequestration to proteasomal degradation has not been explored. We used the yeast Saccharomyces cerevisiae and a model ERAD substrate, the cystic fibrosis transmembrane conductance regulator (CFTR), to explore whether CFTR is sequestered before degradation, to identify the molecular machinery regulating sequestration, and to analyze the relationship between sequestration and degradation. We report that CFTR is sequestered into ER subdomains containing the chaperone Kar2p, and that sequestration and CFTR degradation are disrupted in sec12ts strain (mutant in guanine-nucleotide exchange factor for Sar1p), sec13ts strain (mutant in the Sec13p component of COPII), and sec23ts strain (mutant in the Sec23p component of COPII) grown at restrictive temperature. The function of the Sar1p/COPII machinery in CFTR sequestration and degradation is independent of its role in ER-Golgi traffic. We propose that Sar1p/COPII-mediated sorting of CFTR into ER subdomains is essential for its entry into the proteasomal degradation pathway. These findings reveal a new aspect of the degradative mechanism, and suggest functional crosstalk between the secretory and the degradative pathways. PMID- 12538639 TI - HA95 and LAP2 beta mediate a novel chromatin-nuclear envelope interaction implicated in initiation of DNA replication. AB - HA95 is a chromatin-associated protein that interfaces the nuclear envelope (NE) and chromatin. We report an interaction between HA95 and the inner nuclear membrane protein lamina-associated polypeptide (LAP) 2 beta, and a role of this association in initiation of DNA replication. Precipitation of GST-LAP2 beta fusion proteins and overlays of immobilized HA95 indicate that a first HA95 binding region lies within amino acids 137-242 of LAP2 beta. A second domain sufficient to bind HA95 colocalizes with the lamin B-binding domain of LAP2beta at residues 299-373. HA95-LAP2 beta interaction is not required for NE formation. However, disruption of the association of HA95 with the NH2-terminal HA95-binding domain of LAP2 beta abolishes the initiation, but not elongation, of DNA replication in purified G1 phase nuclei incubated in S-phase extract. Inhibition of replication initiation correlates with proteasome-mediated proteolysis of Cdc6, a component of the prereplication complex. Rescue of Cdc6 degradation with proteasome inhibitors restores replication. We propose that an interaction of LAP2beta, or LAP2 proteins, with HA95 is involved in the control of initiation of DNA replication. PMID- 12538640 TI - The coiled-coil membrane protein golgin-84 is a novel rab effector required for Golgi ribbon formation. AB - Fragmentation of the mammalian Golgi apparatus during mitosis requires the phosphorylation of a specific subset of Golgi-associated proteins. We have used a biochemical approach to characterize these proteins and report here the identification of golgin-84 as a novel mitotic target. Using cryoelectron microscopy we could localize golgin-84 to the cis-Golgi network and found that it is enriched on tubules emanating from the lateral edges of, and often connecting, Golgi stacks. Golgin-84 binds to active rab1 but not cis-Golgi matrix proteins. Overexpression or depletion of golgin-84 results in fragmentation of the Golgi ribbon. Strikingly, the Golgi ribbon is converted into mini-stacks constituting only approximately 25% of the volume of a normal Golgi apparatus upon golgin-84 depletion. These mini-stacks are able to carry out protein transport, though with reduced efficiency compared with a normal Golgi apparatus. Our results suggest that golgin-84 plays a key role in the assembly and maintenance of the Golgi ribbon in mammalian cells. PMID- 12538642 TI - A clathrin/dynamin- and mannose-6-phosphate receptor-independent pathway for granzyme B-induced cell death. AB - The 280-kD cation-independent mannose-6-phosphate receptor (MPR) has been shown to play a role in endocytic uptake of granzyme B, since target cells overexpressing MPR have an increased sensitivity to granzyme B-mediated apoptosis. On this basis, it has been proposed that cells lacking MPR are poor targets for cytotoxic lymphocytes that mediate allograft rejection or tumor immune surveillance. In the present study, we report that the uptake of granzyme B into target cells is independent of MPR. We used HeLa cells overexpressing a dominant-negative mutated (K44A) form of dynamin and mouse fibroblasts overexpressing or lacking MPR to show that the MPR/clathrin/dynamin pathway is not required for granzyme B uptake. Consistent with this observation, cells lacking the MPR/clathrin pathway remained sensitive to granzyme B. Exposure of K44A-dynamin-overexpressing and wild-type HeLa cells to granzyme B with sublytic perforin resulted in similar apoptosis in the two cell populations, both in short and long term assays. Granzyme B uptake into MPR-overexpressing L cells was more rapid than into MPR-null L cells, but the receptor-deficient cells took up granzyme B through fluid phase micropinocytosis and remained sensitive to it. Contrary to previous findings, we also demonstrated that mouse tumor allografts that lack MPR expression were rejected as rapidly as tumors that overexpress MPR. Entry of granzyme B into target cells and its intracellular trafficking to induce target cell death in the presence of perforin are therefore not critically dependent on MPR or clathrin/dynamin-dependent endocytosis. PMID- 12538641 TI - EpsinR: an AP1/clathrin interacting protein involved in vesicle trafficking. AB - EpsinR is a clathrin-coated vesicle (CCV) enriched 70-kD protein that binds to phosphatidylinositol-4-phosphate, clathrin, and the gamma appendage domain of the adaptor protein complex 1 (AP1). In cells, its distribution overlaps with the perinuclear pool of clathrin and AP1 adaptors. Overexpression disrupts the CCV dependent trafficking of cathepsin D from the trans-Golgi network to lysosomes and the incorporation of mannose-6-phosphate receptors into CCVs. These biochemical and cell biological data point to a role for epsinR in AP1/clathrin budding events in the cell, just as epsin1 is involved in the budding of AP2 CCVs. Furthermore, we show that two gamma appendage domains can simultaneously bind to epsinR with affinities of 0.7 and 45 microM, respectively. Thus, potentially, two AP1 complexes can bind to one epsinR. This high affinity binding allowed us to identify a consensus binding motif of the form DFxDF, which we also find in gamma-synergin and use to predict that an uncharacterized EF-hand containing protein will be a new gamma binding partner. PMID- 12538644 TI - Multiple splice variants of the human HIF-3 alpha locus are targets of the von Hippel-Lindau E3 ubiquitin ligase complex. AB - Functional inactivation of the von Hippel-Lindau (VHL) tumor suppressor protein is the cause of familial VHL disease and sporadic kidney cancer. The VHL gene product (pVHL) is a component of an E3 ubiquitin ligase complex that targets the hypoxia-inducible factor (HIF) 1 and 2 alpha subunits for polyubiquitylation. This process is dependent on the hydroxylation of conserved proline residues on the alpha subunits of HIF-1/2 in the presence of oxygen. In our effort to identify orphan HIF-like proteins in the data base that are potential targets of the pVHL complex, we report multiple splice variants of the human HIF-3 alpha locus as follows: hHIF-3 alpha 1, hHIF-3 alpha 2 (also referred to as hIPAS; human inhibitory PAS domain protein), hHIF-3 alpha 3, hHIF-3 alpha 4, hHIF-3 alpha 5, and hHIF-3 alpha 6. We demonstrate that the common oxygen-dependent degradation domain of hHIF-3 alpha 1-3 splice variants is targeted for ubiquitylation by the pVHL complex in vitro and in vivo. This activity is enhanced in the presence of prolyl hydroxylase and is dependent on a proline residue at position 490. Furthermore, the ubiquitin conjugation occurs on lysine residues at position 465 and 568 within the oxygen-dependent degradation domain. These results demonstrate additional targets of the pVHL complex and suggest a growing complexity in the regulation of hypoxia-inducible genes by the HIF family of transcription factors. PMID- 12538645 TI - Compared action of neutrophil proteinase 3 and elastase on model substrates. Favorable effect of S'-P' interactions on proteinase 3 catalysts. AB - Neutrophil proteinase 3 (Pr3) and elastase (NE) may cause lung tissue destruction in emphysema and cystic fibrosis. These serine proteinases have similar P(1) specificities. We have compared their catalytic activity using acyl-tetrapeptide p-nitroanilides, which occupy the S(5)-S'(1) subsites of their substrate binding site, and intramolecularly quenched fluorogenic heptapeptides, which bind at S(5) S'(4). Most p-nitroanilide substrates are turned over slowly by Pr3 as compared with NE. These differences disappear with the fluorogenic heptapeptides, some of which are hydrolyzed even faster by Pr3 than by NE. Elongation of substrates strongly increases the catalytic efficiency of Pr3, whereas it has little effect on NE catalysis. These different sensitivities to S'-P' interactions show that Pr3 and NE are not interchangeable enzymes despite their similar P(1) specificity. PMID- 12538643 TI - RhoA is required for cortical retraction and rigidity during mitotic cell rounding. AB - Mitotic cell rounding is the process of cell shape change in which a flat interphase cell becomes spherical at the onset of mitosis. Rearrangement of the actin cytoskeleton, de-adhesion, and an increase in cortical rigidity accompany mitotic cell rounding. The molecular mechanisms that contribute to this process have not been defined. We show that RhoA is required for cortical retraction but not de-adhesion during mitotic cell rounding. The mitotic increase in cortical rigidity also requires RhoA, suggesting that increases in cortical rigidity and cortical retraction are linked processes. Rho-kinase is also required for mitotic cortical retraction and rigidity, indicating that the effects of RhoA on cell rounding are mediated through this effector. Consistent with a role for RhoA during mitotic entry, RhoA activity is elevated in rounded, preanaphase mitotic cells. The activity of the RhoA inhibitor p190RhoGAP is decreased due to its serine/threonine phosphorylation at this time. Cumulatively, these results suggest that the mitotic increase in RhoA activity leads to rearrangements of the cortical actin cytoskeleton that promote cortical rigidity, resulting in mitotic cell rounding. PMID- 12538646 TI - Peptides targeting caspase inhibitors. AB - Here we report on the identification of peptides targeting the X-inhibitor of apoptosis protein (XIAP). XIAP functions as a caspase inhibitor and is a member of the inhibitors of apoptosis (IAP) family of proteins. IAPs are often overexpressed in cancers and leukemias and are associated with an unfavorable clinical prognosis. We have selected peptides from a phage library by using recombinant full-length human XIAP or a fragment containing only the baculovirus IAP repeat 2 (BIR2) domain. A consensus motif, C(D/E/P)(W/F/Y)-acid/basic-XC, was recovered from two independent screenings by using different libraries. Phage displaying variations of the consensus sequence bound specifically to the BIR2 domain of XIAP but not to other IAPs. The interaction was specific as it could be blocked by the cognate synthetic peptides in a dose-dependent manner. Phage displaying the XIAP-binding motif CEFESC bound to the BIR2 domain of XIAP with an estimated dissociation constant of 1.8 nm as determined by surface plasmon resonance. Protein-protein interaction assays revealed that caspase-3 and caspase 7 (but not caspase-8) blocked the binding of the CEFESC phage to XIAP, indicating that this peptide targets a domain within XIAP that is related to the caspase binding site. In fact, the sequence EFES is homologous to a loop unique to the executioner caspase-3 and caspase-7 that are targeted by XIAP. Finally, we demonstrated that an internalizing version of the XIAP-binding peptide identified in our screenings (PFKQ) can induce programmed cell death in leukemia cells. Peptides interacting with XIAP could serve as prototypes for the design of low molecular weight modulators of apoptosis. PMID- 12538647 TI - X-ray absorption spectroscopy reveals a substantial increase of sulfur oxidation in transthyretin (TTR) upon fibrillization. AB - Transthyretin (TTR) amyloid fibrils are the main component of the amyloid deposits occurring in Familial Amyloidotic Polyneuropathy patients. This is 1 of 20 human proteins leading to protein aggregation disorders such as Alzheimer's and Creutzfeldt-Jakob diseases. The structural details concerning the association of the protein molecules are essential for a better understanding of the disease and consequently the design of new strategies for diagnosis and therapeutics. Disulfide bonds are frequently considered essential for the stability of protein aggregates and since in the TTR monomers there is one cysteine residue, it is important to determine unambiguously the redox state of sulfur present in the fibrils. In this work we used x-ray spectroscopy to further characterize TTR amyloid fibrils. The sulfur K-edge absorption spectra for the wild type and some amyloidogenic TTR variants in the soluble and fibrillar forms were analyzed. Whereas in the soluble proteins the thiol group from cysteine (R-SH) and the thioether group from methionine (R-S-CH(3)) are the most abundant forms, in the TTR fibrils there is a significant oxidation of sulfur to the sulfonate form in the cysteine residue and a partial oxidation of sulfur to sulfoxide in the methionine residues. Further interpretation of the data reveals that there are no disulfide bridges in the fibrillar samples and suggest conformational changes in the TTR molecule, namely in strand A and/or in its vicinity, upon fibril formation. PMID- 12538648 TI - The Methanococcus jannaschii dCTP deaminase is a bifunctional deaminase and diphosphatase. AB - Most bacteria produce the dUMP precursor for thymine nucleotide biosynthesis using two enzymes: a dCTP deaminase catalyzes the formation of dUTP and a dUTP diphosphatase catalyzes pyrophosphate release. Although these two hydrolytic enzymes appear to catalyze very different reactions, they are encoded by homologous genes. The hyperthermophilic archaeon Methanococcus jannaschii has two members of this gene family. One gene, at locus MJ1102, encodes a dUTP diphosphatase, which can scavenge deoxyuridine nucleotides that inhibit archaeal DNA polymerases. The second gene, at locus MJ0430, encodes a novel dCTP deaminase that releases dUMP, ammonia, and pyrophosphate. Therefore this enzyme can singly catalyze both steps in dUMP biosynthesis, precluding the formation of free, mutagenic dUTP. Besides differing from the previously characterized Salmonella typhimurium dCTP deaminase in its reaction products, this archaeal enzyme has a higher affinity for dCTP and its steady-state turnover is faster than the bacterial enzyme. Kinetic studies suggest: 1) the archaeal enzyme specifically recognizes dCTP; 2) dCTP deamination and dUTP diphosphatase activities occur independently at the same active site, and 3) both activities depend on Mg(2+). The bifunctional activity of this M. jannaschii enzyme illustrates the evolution of a suprafamily of related enzymes that catalyze mechanistically distinct reactions. PMID- 12538649 TI - Regulators of G-protein signaling (RGS) 4, insertion into model membranes and inhibition of activity by phosphatidic acid. AB - Regulators of G-protein signaling (RGS) proteins are critical for attenuating G protein-coupled signaling pathways. The membrane association of RGS4 has been reported to be crucial for its regulatory activity in reconstituted vesicles and physiological roles in vivo. In this study, we report that RGS4 initially binds onto the surface of anionic phospholipid vesicles and subsequently inserts into, but not through, the membrane bilayer. Phosphatidic acid, one of anionic phospholipids, could dramatically inhibit the ability of RGS4 to accelerate GTPase activity in vitro. Phosphatidic acid is an effective and potent inhibitor of RGS4 in a G alpha(i1)-[gamma-(32)P]GTP single turnover assay with an IC(50) approximately 4 microm and maximum inhibition of over 90%. Furthermore, phosphatidic acid was the only phospholipid tested that inhibited RGS4 activity in a receptor-mediated, steady-state GTP hydrolysis assay. When phosphatidic acid (10 mol %) was incorporated into m1 acetylcholine receptor-G alpha(q) vesicles, RGS4 GAP activity was markedly inhibited by more than 70% and the EC(50) of RGS4 was increased from 1.5 to 7 nm. Phosphatidic acid also induced a conformational change in the RGS domain of RGS4 measured by acrylamide-quenching experiments. Truncation of the N terminus of RGS4 (residues 1-57) resulted in the loss of both phosphatidic acid binding and lipid-mediated functional inhibition. A single point mutation in RGS4 (Lys(20) to Glu) permitted its binding to phosphatidic acid-containing vesicles but prevented lipid-induced conformational changes in the RGS domain and abolished the inhibition of its GAP activity. We speculate that the activation of phospholipase D or diacylglycerol kinase via G protein mediated signaling cascades will increase the local concentration of phosphatidic acid, which in turn block RGS4 GAP activity in vivo. Thus, RGS4 may represent a novel effector of phosphatidic acid, and this phospholipid may function as a feedback regulator in G protein-mediated signaling pathways. PMID- 12538650 TI - A developmentally regulated splice variant from the complex lola locus encoding multiple different zinc finger domain proteins interacts with the chromosomal kinase JIL-1. AB - Using a yeast two-hybrid screen we have identified a novel isoform of the lola locus, Lola zf5, that interacts with the chromosomal kinase JIL-1. We characterized the lola locus and provide evidence that it is a complex locus from which at least 17 different splice variants are likely to be generated. Fifteen of these each have a different zinc finger domain, whereas two are without. This potential for expression of multiple gene products suggests that they serve diverse functional roles in different developmental contexts. By Northern and Western blot analyses we demonstrate that the expression of Lola zf5 is developmentally regulated and that it is restricted to early embryogenesis. Immunocytochemical labeling with a Lola zf5-specific antibody of Drosophila embryos indicates that Lola zf5 is localized to nuclei. Furthermore, by creating double-mutant flies we show that a reduction of Lola protein levels resulting from mutations in the lola locus acts as a dominant modifier of a hypomorphic JIL 1 allele leading to an increase in embryonic viability. Thus, genetic interaction assays provide direct evidence that gene products from the lola locus function within the same pathway as the chromosomal kinase JIL-1. PMID- 12538651 TI - Type I collagen triplet duplication mutation in lethal osteogenesis imperfecta shifts register of alpha chains throughout the helix and disrupts incorporation of mutant helices into fibrils and extracellular matrix. AB - The majority of collagen mutations causing osteogenesis imperfecta (OI) are glycine substitutions that disrupt formation of the triple helix. A rare type of collagen mutation consists of a duplication or deletion of one or two Gly-X-Y triplets. These mutations shift the register of collagen chains with respect to each other in the helix but do not interrupt the triplet sequence, yet they have severe clinical consequences. We investigated the effect of shifting the register of the collagen helix by a single Gly-X-Y triplet on collagen assembly, stability, and incorporation into fibrils and matrix. These studies utilized a triplet duplication in COL1A1 exon 44 that occurred in the cDNA and gDNA of two siblings with lethal OI. The normal allele encodes three identical Gly-Ala-Hyp triplets at aa 868-876, whereas the mutant allele encodes four. The register shift delays helix formation, causing overmodification. Differential scanning calorimetry yielded a decrease in T(m) of 2 degrees C for helices with one mutant chain and a 6 degrees C decrease in helices with two mutant chains. An in vitro binary co-processing assay of N-proteinase cleavage demonstrated that procollagen with the triplet duplication has slower N-propeptide cleavage than in normal controls or procollagen with proalpha1(I) G832S, G898S, or G997S substitutions, showing that the register shift persists through the entire helix. The register shift disrupts incorporation of mutant collagen into fibrils and matrix. Proband fibrils formed inefficiently in vitro and contained only normal helices and helices with a single mutant chain. Helices with two mutant chains and a significant portion of helices with one mutant chain did not form fibrils. In matrix deposited by proband fibroblasts, mutant chains were abundant in the immaturely cross-linked fraction but constituted a minor fraction of maturely cross-linked chains. The profound effects of shifting the collagen triplet register on chain interactions in the helix and on fibril formation correlate with the severe clinical consequences. PMID- 12538652 TI - Regulation of biglycan gene expression by transforming growth factor-beta requires MKK6-p38 mitogen-activated protein Kinase signaling downstream of Smad signaling. AB - Several signaling pathways have been implicated in mediating TGF-beta1-induced extracellular matrix production and fibrosis. We have shown recently that induction of biglycan (BGN) expression by TGF-beta1 depended on a functional Smad pathway (Chen, W.-B., Lenschow, W., Tiede, K., Fischer, J. W., Kalthoff, H., and Ungefroren, H. (2002) J. Biol. Chem. 277, 36118-36128). Here, we present evidence that the ability of TGF-beta 1 to induce BGN mRNA, in addition to Smads, requires p38 MAPK signaling, because 1) pharmacological inhibitors of p38 dose-dependently inhibited the TGF-beta effect without significantly affecting the transcriptional activity of a constitutively active mutant of the TGF-beta type I receptor or Smad2 phosphorylation at concentrations up to 10 microm, 2) the up-regulation of BGN mRNA was preceded by a delayed increase in the phosphorylation of p38 and its upstream activator MKK6 in TGF-beta 1-treated PANC-1 cells, 3) inhibition of the p38 pathway by stable retroviral transduction with a dominant negative mutant of either p38 or MKK6 reduced TGF-beta 1-induced BGN mRNA expression, and 4) overexpression of wild-type p38 or MKK6, but not MKK3, augmented the TGF-beta 1 effect on BGN mRNA. We further demonstrate that the (delayed) p38 activation by TGF-beta 1 is downstream of Smads and requires a functional Smad pathway, because blocking TGF-beta-induced p38 activity with SB202190 had no effect on Smad2 phosphorylation, but blocking Smad signaling by forced expression of Smad7 abolished TGF-beta1 induction of p38 activation and, as shown earlier, BGN mRNA expression; finally, re-expression of Smad4 in Smad4-null CFPAC-1 cells restored TGF-beta-induced p38 phosphorylation and, as demonstrated previously, BGN mRNA accumulation. These results clearly show that TGF-beta induction of BGN expression in pancreatic cells requires activation of MKK6-p38 MAPK signaling downstream of Smad signaling and provide a mechanistic clue to the up-regulation of BGN seen in inflammatory response-related fibrosis and desmoplasia. PMID- 12538653 TI - Inhibition of T cell function during malaria: implications for immunology and vaccinology. PMID- 12538654 TI - Malaria blood stage suppression of liver stage immunity by dendritic cells. AB - Malaria starts with Plasmodium sporozoites infection of the host's liver, where development into blood stage parasites occurs. It is not clear why natural infections do not induce protection against the initial liver stage and generate low CD8+ T cell responses. Using a rodent malaria model, we show that Plasmodium blood stage infection suppresses CD8+ T cell immune responses that were induced against the initial liver stage. Blood stage Plasmodium affects dendritic cell (DC) functions, inhibiting maturation and the capacity to initiate immune responses and inverting the interleukin (IL)-12/IL-10 secretion pattern. The interaction of blood stage parasites with DCs induces the secretion of soluble factors that inhibit the activation of CD8+ T cells in vitro and the suppression of protective CD8+ T cell responses against the liver stage in vivo. We propose that blood stage infection induces DCs to suppress CD8+ T cell responses in natural malaria infections. This evasion mechanism leaves the host unprotected against reinfection by inhibiting the immune response against the initial liver stage of the disease. PMID- 12538655 TI - Vaginal submucosal dendritic cells, but not Langerhans cells, induce protective Th1 responses to herpes simplex virus-2. AB - Herpes simplex virus (HSV) type 2 infection occurs primarily at the genital mucosal surfaces and is a leading cause of ulcerative lesions. Despite the availability of animal models for HSV-2 infection, little is known regarding the mechanism of immune induction within the vaginal mucosa. Here, we examined the cell types responsible for the initiation of protective Th1 immunity to HSV-2. Intravaginal inoculation of HSV-2 led to a rapid recruitment of submucosal dendritic cells (DCs) to the infected epithelium. Subsequently, CD11c(+) DCs harboring viral peptides in the context of MHC class II molecules emerged in the draining lymph nodes and were found to be responsible for the stimulation of IFNgamma secretion from HSV-specific CD4(+) T cells. Other antigen-presenting cells including B cells and macrophages did not present viral peptides to T cells in the draining lymph nodes. Next, we assessed the relative contribution to immune generation by the Langerhans cells in the vaginal epithelium, the submucosal CD11b(+) DCs, and the CD8alpha(+) lymph node DCs. Analysis of these DC populations from the draining lymph nodes revealed that only the CD11b(+) submucosal DCs, but not Langerhans cell-derived or CD8alpha(+) DCs, presented viral antigens to CD4(+) T cells and induced IFNgamma secretion. These results demonstrate a previously unanticipated role for submucosal DCs in the generation of protective Th1 immune responses to HSV-2 in the vaginal mucosa, and suggest their importance in immunity to other sexually transmitted diseases. PMID- 12538656 TI - Human T cell receptor gammadelta cells recognize endogenous mevalonate metabolites in tumor cells. AB - T lymphocytes expressing the T cell receptor (TCR)-gammadelta recognize unknown antigens on tumor cells. Here we identify metabolites of the mevalonate pathway as the tumor ligands that activate TCR-gammadelta cells. In tumor cells, blockade of hydroxy-methylglutaryl-CoA reductase (HMGR), the rate limiting enzyme of the mevalonate pathway, prevents both accumulation of mevalonate metabolites and recognition by TCR-gammadelta cells. When metabolite accumulation is induced by overexpressing HMGR or by treatment with nitrogen-containing bisphosphonate drugs, tumor cells derived from many tissues acquire the capacity to stimulate the same TCR-gammadelta population. Accumulation of mevalonate metabolites in tumor cells is a powerful danger signal that activates the immune response and may represent a novel target of tumor immunotherapy. PMID- 12538657 TI - Differential regulation of cathepsin S and cathepsin L in interferon gamma treated macrophages. AB - Cathepsin S (catS) and cathepsin L (catL) mediate late stages of invariant chain (Ii) degradation in discrete antigen-presenting cell types. Macrophages (Mphis) are unique in that they express both proteases and here we sought to determine the relative contribution of each enzyme. We observe that catL plays no significant role in Ii cleavage in interferon (IFN)-gamma-stimulated Mphis. In addition, our studies show that the level of catL activity is significantly decreased in Mphis cultured in the presence of IFN-gamma whereas catS activity increases. The decrease in catL activity upon cytokine treatment occurs despite the persistence of high levels of mature catL protein, suggesting that a specific inhibitor of the enzyme is up-regulated in IFN-gamma-stimulated peritoneal Mphis. Similar inhibition of activity is observed in dendritic cells engineered to overexpress catL. Such enzymatic inhibition in Mphis exhibits only partial dependence upon Ii and therefore, other mechanisms of catL inhibition are regulated by IFN-gamma. Thus, during a T helper cell type 1 immune response catL inhibition in Mphis results in preferential usage of catS, such that major histocompatibility complex class II presentation by all bone marrow-derived antigen-presenting cell is regulated by catS. PMID- 12538658 TI - Expression of ICOS in vivo defines CD4+ effector T cells with high inflammatory potential and a strong bias for secretion of interleukin 10. AB - The studies performed to date analyzed the overall participation of the inducible costimulator (ICOS) in model diseases, but did not yield information on the nature and function of ICOS-expressing T cells in vivo. We examined ICOS(+) T cells in the secondary lymphoid organs of nonmanipulated mice, in the context of an "unbiased" immune system shaped by environmental antigens. Using single cell analysis, ICOS(low) cells were found to be loosely associated with the early cytokines interleukin (IL)-2, IL-3, IL-6, and interferon (IFN)-gamma. ICOS(medium) cells, the large majority of ICOS(+) T cells in vivo, were very tightly associated with the synthesis of the T helper type 2 (Th2) cytokines IL 4, IL-5, and IL-13, and these cells exhibited potent inflammatory effects in vivo. In contrast, ICOS(high) T cells were highly and selectively linked to the anti-inflammatory cytokine IL-10. Overall, these data seem to indicate that ICOS cell surface density serves as a regulatory mechanism for the release of cytokines with different immunological properties. Further in vivo functional experiments with in vitro-activated T cells strongly suggested that the ICOS(+) population, although representing in vivo only around 10% of T cells bearing early or late activation markers, nevertheless encompasses virtually all effector T cells, a finding with major diagnostic and therapeutic implications. PMID- 12538659 TI - T cell accumulation in B cell follicles is regulated by dendritic cells and is independent of B cell activation. AB - We investigated the mechanism of CD4 T cell accumulation in B cell follicles after immunization. Follicular T cell numbers were correlated with the number of B cells, indicating B cell control of the niche that T cells occupy. Despite this, we found no role for B cells in the follicular migration of T cells. Instead, T cells are induced to migrate into B cell follicles entirely as a result of interaction with dendritic cells (DCs). Migration relies on CD40 dependent maturation of DCs, as it did not occur in CD40-deficient mice but was reconstituted with CD40(+) DCs. Restoration was not achieved by the activation of DCs with bacterial activators (e.g., lipopolysaccharide, CpG), but was by the injection of OX40L-huIgG1 fusion protein. Crucially, the up-regulation of OX40L (on antigen-presenting cells) and CXCR-5 (on T cells) are CD40-dependent events and we show that T cells do not migrate to follicles in immunized OX40-deficient mice. PMID- 12538660 TI - Prospective estimation of recombination signal efficiency and identification of functional cryptic signals in the genome by statistical modeling. AB - The recombination signals (RS) that guide V(D)J recombination are phylogenetically conserved but retain a surprising degree of sequence variability, especially in the nonamer and spacer. To characterize RS variability, we computed the position-wise information, a measure correlated with sequence conservation, for each nucleotide position in an RS alignment and demonstrate that most position-wise information is present in the RS heptamers and nonamers. We have previously demonstrated significant correlations between RS positions and here show that statistical models of the correlation structure that underlies RS variability efficiently identify physiologic and cryptic RS and accurately predict the recombination efficiencies of natural and synthetic RS. In scans of mouse and human genomes, these models identify a highly conserved family of repetitive DNA as an unexpected source of frequent, cryptic RS that rearrange both in extrachromosomal substrates and in their genomic context. PMID- 12538661 TI - Host prostaglandin E(2)-EP3 signaling regulates tumor-associated angiogenesis and tumor growth. AB - Nonsteroidal antiinflammatories are known to suppress incidence and progression of malignancies including colorectal cancers. However, the precise mechanism of this action remains unknown. Using prostaglandin (PG) receptor knockout mice, we have evaluated a role of PGs in tumor-associated angiogenesis and tumor growth, and identified PG receptors involved. Sarcoma-180 cells implanted in wild-type (WT) mice formed a tumor with extensive angiogenesis, which was greatly suppressed by specific inhibitors for cyclooxygenase (COX)-2 but not for COX-1. Angiogenesis in sponge implantation model, which can mimic tumor-stromal angiogenesis, was markedly suppressed in mice lacking EP3 (EP3(-/-)) with reduced expression of vascular endothelial growth factor (VEGF) around the sponge implants. Further, implanted tumor growth (sarcoma-180, Lewis lung carcinoma) was markedly suppressed in EP3(-/-), in which tumor-associated angiogenesis was also reduced. Immunohistochemical analysis revealed that major VEGF-expressing cells in the stroma were CD3/Mac-1 double-negative fibroblasts, and that VEGF expression in the stroma was markedly reduced in EP3(-/-), compared with WT. Application of an EP3 receptor antagonist inhibited tumor growth and angiogenesis in WT, but not in EP3(-/-). These results demonstrate significance of host stromal PGE(2)-EP3 receptor signaling in tumor development and angiogenesis. An EP3 receptor antagonist may be a candidate of chemopreventive agents effective for malignant tumors. PMID- 12538662 TI - Unique resistance of I/LnJ mice to a retrovirus is due to sustained interferon gamma-dependent production of virus-neutralizing antibodies. AB - Selection of immune escape variants impairs the ability of the immune system to sustain an efficient antiviral response and to control retroviral infections. Like other retroviruses, mouse mammary tumor virus (MMTV) is not efficiently eliminated by the immune system of susceptible mice. In contrast, MMTV-infected I/LnJ mice are capable of producing IgG2a virus-neutralizing antibodies, sustain this response throughout their life, and secrete antibody-coated virions into the milk, thereby preventing infection of their progeny. Antibodies were produced in response to several MMTV variants and were cross-reactive to them. Resistance to MMTV infection was recessive and was dependent on interferon (IFN)-gamma production, because I/LnJ mice with targeted deletion of the INF-gamma gene failed to produce any virus-neutralizing antibodies. These findings reveal a novel mechanism of resistance to retroviral infection that is based on a robust and sustained IFN-gamma-dependent humoral immune response. PMID- 12538663 TI - DNA methylation maintains allele-specific KIR gene expression in human natural killer cells. AB - Killer immunoglobulin-like receptors (KIR) bind self-major histocompatibility complex class I molecules, allowing natural killer (NK) cells to recognize aberrant cells that have down-regulated class I. NK cells express variable numbers and combinations of highly homologous clonally restricted KIR genes, but uniformly express KIR2DL4. We show that NK clones express both 2DL4 alleles and either one or both alleles of the clonally restricted KIR 3DL1 and 3DL2 genes. Despite allele-independent expression, 3DL1 alleles differed in the core promoter by only one or two nucleotides. Allele-specific 3DL1 gene expression correlated with promoter and 5' gene DNA hypomethylation in NK cells in vitro and in vivo. The DNA methylase inhibitor, 5-aza-2'-deoxycytidine, induced KIR DNA hypomethylation and heterogeneous expression of multiple KIR genes. Thus, NK cells use DNA methylation to maintain clonally restricted expression of highly homologous KIR genes and alleles. PMID- 12538664 TI - A single amino acid alteration in cytoplasmic domain determines IL-2 promoter activation by ligation of CD28 but not inducible costimulator (ICOS). AB - The CD28 family molecules, CD28, and inducible costimulator (ICOS) all provide positive costimulatory signals. However, unlike CD28, ICOS does not costimulate IL-2 secretion. The YMNM motif that exists in the CD28 cytoplasmic domain is a known binding site for phosphatidylinositol 3-kinase (PI3-K) and Grb2. ICOS possesses the YMFM motif in the corresponding region of CD28 that binds PI3-K but not Grb2. We postulated that the reason that ICOS does not have the ability to induce IL-2 production is because it fails to recruit Grb2. To verify this hypothesis, we generated a mutant ICOS gene that contains the CD28 YMNM motif and measured IL-2 promoter activation after ICOS ligation. The results indicated that ICOS became competent to activate the IL-2 promoter by this single alteration. Further analysis demonstrated that Grb2 binding to ICOS was sufficient to activate the NFAT/AP-1 site in the IL-2 promoter and that the cytoplasmic domain of CD28 outside of the YMNM motif is required for activation of the CD28RE/AP-1 and NF-kappaB sites. Together, these observations lead us to believe that the difference of a single amino acid, which affects Grb2 binding ability, may define a functional difference between the CD28- and ICOS-mediated costimulatory signals. PMID- 12538665 TI - Inhibition of interleukin 1 receptor/Toll-like receptor signaling through the alternatively spliced, short form of MyD88 is due to its failure to recruit IRAK 4. AB - Toll-like receptors (TLRs) and members of the proinflammatory interleukin 1 receptor (IL-1R) family are dependent on the presence of MyD88 for efficient signal transduction. The bipartite nature of MyD88 (N-terminal death domain [DD] and COOH-terminal Toll/IL-1 receptor [TIR] domain) allows it to link the TIR domain of IL-1R/TLR with the DD of the Ser/Thr kinase termed IL-1R-associated kinase (IRAK)-1. This triggers IRAK-1 phosphorylation and in turn the activation of multiple signaling cascades such as activation of the transcription factor nuclear factor (NF)-kappaB. In contrast, expression of MyD88 short (MyD88s), an alternatively spliced form of MyD88 that lacks only the short intermediate domain separating the DD and TIR domains, leads to a shutdown of IL-1/lipopolysaccharide induced NF-kappaB activation. Here, we provide the molecular explanation for this difference. MyD88 but not MyD88s strongly interacts with IRAK-4, a newly identified kinase essential for IL-1R/TLR signaling. In the presence of MyD88s, IRAK-1 is not phosphorylated and neither activates NF-kappaB nor is ubiquitinated. Thus, MyD88s acts as a negative regulator of IL-1R/TLR/MyD88 triggered signals, leading to a transcriptionally controlled negative regulation of innate immune responses. PMID- 12538667 TI - Cutting edge: IFN consensus sequence binding protein/IFN regulatory factor 8 drives the development of type I IFN-producing plasmacytoid dendritic cells. AB - IFN consensus sequence binding protein (ICSBP/IFN regulatory factor 8) is a hematopoietic cell-specific transcription factor essential for the generation of CD8 alpha(+) dendritic cells (DCs). We found that ICSBP(-/-) mice lack B220(+)CD11b(-) plasmacytoid DCs (pDCs) in addition to CD8 alpha(+) DCs. Although ICSBP(-/-) mice have B220(-)CD11b(+) myeloid DCs (mDCs), they fail to mature upon Toll-like receptor signaling. Accordingly, ICSBP(-/-) bone marrow progenitor cells were defective in generating pDCs in the fms-like tyrosine kinase 3 ligand based culture system and mDCs generated in this system were defective in maturation. We demonstrate that introduction of ICSBP rescues the development of pDCs from -/- bone marrow progenitors. ICSBP also restored the ability of both pDCs and mDCs to mature after Toll-like receptor signals. ICSBP-restored DCs produced IFN-alpha and IL-12p40 in a DC subset-selective manner with the amounts comparable to those by +/+ DCs. Together, ICSBP is essential for early pDC development and final maturation of both pDCs and mDCs. PMID- 12538668 TI - Cutting edge: profile of chemokine receptor expression on human plasma cells accounts for their efficient recruitment to target tissues. AB - We systematically examined the repertoire of chemokine receptors expressed by human plasma cells. Fresh bone marrow plasma cells and myeloma cells consistently expressed CXCR4, CXCR6, CCR10, and CCR3. Accordingly, plasma cells responded to their respective ligands in chemotaxis and very late Ag-4-dependent cell adhesion to fibronectin. Immobilized CXC chemokine ligand (CXCL)16, a novel transmembrane type chemokine and CXCR6 ligand, also directly induced adhesion of plasma cells without requiring G(alpha i) signaling or divalent cations. Furthermore, we revealed consistent expression of CXCL12 (CXCR4 ligand), CXCL16 (CXCR6 ligand), and CC chemokine ligand 28 (CCR10 and CCR3 ligand) in tissues enriched with plasma cells including bone marrow, and constitutive expression of CXCL12, CXCL16, and CC chemokine ligand 28 by cultured human bone marrow stromal cells. Collectively, plasma cells are likely to be recruited to bone marrow and other target tissues via CXCR4, CXCR6, CCR10, and CCR3. CXCR6 may also contribute to tissue localization of plasma cells through its direct binding to membrane anchored CXCL16. PMID- 12538669 TI - Transgenic expression of RasGRP1 induces the maturation of double-negative thymocytes and enhances the production of CD8 single-positive thymocytes. AB - RasGRP1 is a guanine nucleotide exchange factor for Ras that is required for the efficient production of both CD4 and CD8 single-positive thymocytes. We found that RasGRP1 expression is rapidly up-regulated in double-negative thymocytes following pre-TCR ligation. Transgenic overexpression of RasGRP1 compensated for deficient pre-TCR signaling in vivo, enabling recombinase-activating gene 2(-/-) double-negative thymocytes to mature to the double-positive stage. RasGRP1 transgenic mice had a 4-fold increase in CD8 single-positive thymocytes, most of which had atypically low levels of CD3. The RasGRP1 transgene lowered the threshold of TCR signaling needed to initiate proliferation of single-positive thymocytes, with this effect being particularly evident among CD8 single-positive cells. In 3-day cultures, TCR stimulation via anti-CD3 caused a 10-fold increase in the ratio of CD8 to CD4 thymocytes among RasGRP1 transgenic vs nontransgenic thymocytes. These results demonstrate that in addition to driving the double negative to double-positive transition, increased expression of RasGRP1 selectively increases CD8 single-positive thymocyte numbers and enhances their responsiveness to TCR signaling. PMID- 12538670 TI - Deregulated MHC class II transactivator expression leads to a strong Th2 bias in CD4+ T lymphocytes. AB - The MHC class II (MHC-II) transactivator (CIITA) is the master transcriptional regulator of genes involved in MHC-II-restricted Ag presentation. Fine tuning of CIITA gene expression determines the cell type-specific expression of MHC-II genes. This regulation is achieved by the selective usage of multiple CIITA promoters. It has recently been suggested that CIITA also contributes to Th cell differentiation by suppressing IL-4 expression in Th1 cells. In this study, we show that endogenous CIITA is expressed at low levels in activated mouse T cells. Importantly CIITA is not regulated differentially in murine and human Th1 and Th2 cells. Ectopic expression of a CIITA transgene in multiple mouse cell types including T cells, does not interfere with normal development of CD4(+) T cells. However, upon TCR activation the CIITA transgenic CD4(+) T cells preferentially differentiate into IL-4-secreting Th2-type cells. These results imply that CIITA is not a direct Th1-specific repressor of the IL-4 gene and that tight control over the expression of CIITA and MHC-II is required to maintain the normal balance between Th1 and Th2 responses. PMID- 12538671 TI - Comparative effects of human Ig alpha and Ig beta in inducing autoreactive antibodies against B cells in mice. AB - Human and mouse Ig alpha molecules share only 58% amino acid sequence identity in their extracellular regions. However, mice immunized with a recombinant Fc fusion protein containing the extracellular portion of human Ig alpha produced significant amounts of IgG capable of binding to Ig alpha on mouse B cells. The induced auto/cross-reactive Abs could down-regulate B cell levels and the consequent humoral immune responses against an irrelevant Ag in treated mice. Analogous immunization with an Fc fusion protein containing the extracellular portion of human Ig beta gave a much weaker response to mouse Ig beta, although human and mouse Ig beta, like their Ig alpha counterparts, share 56% sequence identity in their extracellular regions. Protein sequence analyses indicated that a potential immunogenic segment, located at the C-terminal loop of the extracellular domain, has an amino acid sequence that is identical between human and mouse Ig alpha. A mAb A01, which could bind to both human and mouse Ig alpha, was found to be specific to a peptide encompassing this immunogenic segment. These findings suggest that specific auto/cross-reactivity against self Ig alpha can be induced by a molecular mimicry presented by a foreign Ig alpha. PMID- 12538672 TI - Sustained depolarization and ADP-ribose activate a common ionic current in rat peritoneal macrophages. AB - Phagocytosis is associated with large changes in the membrane potential of macrophages, but the functional significance of this is unknown. Whole cell recordings were made from rat peritoneal macrophages. Sustained (>30 s) depolarization of the cells progressively activated a conductance that remained high (several nanoSeimens) for several tens of seconds. This current: 1) was linearly dependent on potential between -100 and +50 mV; 2) reversed close to 0 mV in a physiological external solution; 3) could also be carried in part by N methyl-D-glucamine (P(NMDG)/P(Na) 0.7), chloride (P(Cl)/P(Na) 0.5), or calcium (P(Ca)/P(Na) 1.3); and 4) was blocked by intracellular ATP (5 mM) or ADP (10 mM) and by extracellular lanthanum (half-maximal concentration 1 mM). A current with all the same properties was recorded in cells when the intracellular solution contained ADP-ribose (10-300 micro M) or beta-NAD (1 mM) (but not any other nucleotide analogs tested). The results suggest that prolonged depolarization leads to an increased intracellular level of ADP-ribose, which in turn activates this nonselective conductance(s). PMID- 12538673 TI - IFN-alpha beta released by Mycobacterium tuberculosis-infected human dendritic cells induces the expression of CXCL10: selective recruitment of NK and activated T cells. AB - We recently reported that dendritic cells (DC) infected with Mycobacterium tuberculosis (Mtb) produce Th1/IFN-gamma-inducing cytokines, IFN-alpha beta and IL-12. In the present article, we show that maturing Mtb-infected DC express high levels of CCR7 and they become responsive to its ligand CCL21. Conversely, CCR5 expression was rapidly lost from the cell surface following Mtb infection. High levels of CCL3 and CCL4 were produced within 8 h after infection, which is likely to account for the observed CCR5 down-modulation on Mtb-infected DC. In addition, Mtb infection stimulated the secretion of CXCL9 and CXCL10. Interestingly, the synthesis of CXCL10 was mainly dependent on the Mtb-induced production of IFN alpha beta. Indeed, IFN-alpha beta neutralization down-regulated CXCL10 expression, whereas the expression of CXCL9 appeared to be unaffected. The chemotactic activity of the Mtb-infected DC supernatants was evaluated by migration assays using activated NK, CD4(+), and CD8(+) cells that expressed both CCR5 and CXCR3. Mtb-induced expression of CCL3, CCL4, CXCL9, and CXCL10 was involved in the stimulation of NK and T cell migration. In accordance with the data on the IFN-alpha beta-induced expression of CXCL10, neutralization of IFN alpha beta significantly reduced the chemotactic activity of the supernatant from Mtb-infected DC. This indicates that IFN-alpha beta may modulate the immune response through the expression of CXCL10, which along with CXCL9, CCL3, and CCL4 participates in the recruitment and selective homing of activated/effector cells, which are known to accumulate at the site of Mtb infection and take part in the formation of the granulomas. PMID- 12538674 TI - Tumor eradication by hepatitis B virus X antigen-specific CD8+ T cells in xenografted nude mice. AB - We have previously reported several CTL epitopes derived from the hepatitis B viral X Ag (HBx). In this study, we evaluated whether HBx-specific CTLs can be effectively used in adoptive cancer immunotherapy. To validate the possibility, four peptides containing a HLA-A2.1-restricted binding consensus motif were identified from the HBx protein and tested for their ability to activate CTL from PBMCs isolated from chronic carriers of HBV (n = 12). We selected two highly potent epitopes, HBx 52-60 (HLSLRGLFV) and HBx 115-123 (CLFKDWEEL), that are capable of inducing Ag-specific cytotoxic T cells in patient PBMCs. For adoptive immunotherapy using HBx-specific CTLs, we generated CTL clones restricted to the HBx 52-60 or HBx 115-123 peptide using a limiting dilution technique. LC-46, an HBx 52-60-specific clone, is CD62L(-)CD69(+)CD45RO(+)CD45RA(-)CD25(dim) and is stained by IFN-gamma (approximately 92%), IL-2 (30%), and TNF-alpha (56%), but not by IL-5, IL-10, IL-12, or TNF-beta, indicating that the cells are fully activated T cytotoxic 1-type cells. When LC-46 cells were adoptively transferred into xenografted nude mice bearing human hepatomas expressing HLA-A2.1 molecules and intracellular HBx proteins, the tumors were eradicated. Taken together, our data provide solid evidence for the feasibility of adoptive immunotherapy with HBx-sensitized CTLs in hepatitis disease, including hepatocellular carcinoma (HCC). PMID- 12538675 TI - Cross-presentation of HLA class I epitopes from exogenous NY-ESO-1 polypeptides by nonprofessional APCs. AB - NY-ESO-1, a germ cell Ag often detected in tumor tissues, frequently elicits Ab and CD8(+) T cell responses in cancer patients. Overlapping long peptides spanning the NY-ESO-1 sequence have been used to map HLA class I-restricted epitopes recognized by NY-ESO-1-specific CD8(+) T lymphocytes. To address the antigenicity of long peptides, we analyzed two synthetic 30-mer peptides from NY ESO-1, polypeptides 80-109 and 145-174, for their capacity to be processed by APCs and to stimulate CD8(+) T cells. By incubating APCs with polypeptides at different temperatures or in the presence of protease inhibitors, we found that NY-ESO-1 polypeptides were rapidly internalized by B cells, T2 cells, or PBLs and submitted to cellular proteolytic action to yield nonamer epitopes presented by HLA class I. Polypeptides were also immunogenic in vitro and stimulated the expansion of CD8(+) T cells against naturally processed NY-ESO-1 epitopes in the context of three different HLA class I alleles. Polypeptides can thus serve as exogenous Ags that are cross-presented on HLA class I without requiring the action of professional APCs. These findings support innovative vaccination strategies using NY-ESO-1 polypeptides that would circumvent current limitations of HLA class I peptide vaccination, i.e., HLA eligibility criteria and knowledge of epitope, while allowing for facilitated immunogenicity in the presence of helper epitopes. PMID- 12538676 TI - The contribution of NKT cells, NK cells, and other gamma-chain-dependent non-T non-B cells to IL-12-mediated rejection of tumors. AB - IL-12 is a potent cytokine that impairs the growth of several tumors in vivo in natural as well as in therapeutic conditions. Although IL-12 can enhance a number of immunological antitumor mechanisms, including those mediated by NK cells and CTL, recent reports have suggested that the mouse CD1d-restricted V alpha 14-J alpha 18 NKT cell was the essential cell type recruited in most, if not all tumor rejection models, including the B16 melanoma. In this study, we have examined and compared the role of NKT cells, T cells, NK cells, and other non-T non-B cells in the rejection of B16 melanoma cells after exogenous administration of IL-12. Surprisingly, our results failed to confirm a necessary role for NKT cells in this model. Instead, we found that NK cells mediated the rejection of liver metastases, whereas other gamma c-dependent non-T non-B cells, possibly lymphoid dendritic cells, were required for rejection of skin tumors. These findings challenge the view that NKT cells are systematically required for IL-12-mediated rejection of tumors, and instead reveal that a variety of effector pathways can be recruited depending on the tumor microenvironment. PMID- 12538677 TI - Broadening of epitope recognition during immune rejection of ErbB-2-positive tumor prevents growth of ErbB-2-negative tumor. AB - Tumor heterogeneity is a limiting factor in Ag-specific vaccination. Ag-negative variants may arise after tumor cells bearing the immunizing Ags are destroyed. In situ priming to tumor-associated epitopes distinct from and not cross-reactive with the immunizing Ags may be crucial to the ultimate success of cancer vaccination. Immunization of BALB/c mice with DNA encoding wild-type human ErbB-2 (Her-2/neu, E2) or cytoplasmic ErbB-2 (cytE2), activated primarily CD4 or CD8 T cells, respectively, and both vaccines protected against ErbB-2-positive D2F2/E2 tumors. In > or =50% of protected mice, a second challenge of ErbB-2-negative D2F2 tumor cells was rejected. Recognition of non-ErbB-2, tumor-associated Ags was demonstrated by immune cell proliferation upon stimulation with irradiated D2F2 cells. This broadening of epitope recognition was abolished if CD4 T cells were depleted before D2F2/E2 tumor challenge, demonstrating their critical role in Ag priming. Similarly, mice that rejected D2F2/cytE2 tumor cells, which express only MHC I epitopes of ErbB-2, were not protected from a second challenge with D2F2 cells. Depletion of CD8 T cells abolished protection against D2F2, indicating the activation of D2F2-specific CTL. Therefore, long term protection may be achieved by immunization with dominant Ag(s), followed by a general enhancement of CD4 T cell activity to promote priming to multiple tumor associated Ags. PMID- 12538678 TI - Transfer of TCR genes into mature T cells is accompanied by the maintenance of parental T cell avidity. AB - The adoptive transfer of tumor-specific T cells expanded in vitro can be of significant therapeutic value in select cancer patients. This strategy is limited though, as it is often difficult, if not impossible, to obtain T cells of clinical value. The transfer of TCR genes to mature T cells to generate tumor reactive T cells provides a potential mechanism to overcome these limitations. To evaluate the feasibility of such an approach and the quality of the resulting T cells, we generated replication-deficient retroviral vectors using the well characterized OT-1 TCR genes. After transducing murine T cells, we were able to expand large numbers of Ag-specific T cells that were functionally active against tumor cells expressing the relevant Ag. Furthermore, we found that T cells expressing retrovirally encoded TCR had avidity that was similar to that of the parental clone. This maintenance of avidity was despite variable expression of the retrovirally encoded TCR and the presence of potentially competing endogenous TCRs. These results suggest that the inherent qualities of the TCR, as dictated by the coding sequence, are the most critical parameters in the generation of high-avidity T cells. PMID- 12538679 TI - Differential antigen sensitivity and costimulatory requirements in human Th1 and Th2 antigen-specific CD4+ cells with similar TCR avidity. AB - The differentiation of naive CD4(+) Th cells into Th1 and Th2 phenotypes is influenced by cytokines, concentration of Ag, accessory molecules, and the affinity of the MHC-TCR interaction. To study these factors in human memory T cells, T cell lines with Th1 or Th2 phenotypes specific for the peptide hemagglutinin (HA)(307-319) in the context of DRB1*0401 were established from the peripheral blood of an individual previously vaccinated for influenza virus. Flow cytometric analysis with fluorescent-labeled MHC class II tetramers was used to analyze TCR avidity: the Th2 line bound the HLA-DR*0401-HA(307-319) tetramers with higher mean avidity, although the range of binding avidity largely overlapped with the Th1 line. High-affinity Th1 and Th2 lines were established for further study by FACS sorting. When activated with plate-bound HLA-DR*0401 HA(307-319) monomers, the Th1 line proliferated and produced IFN-gamma without additional costimulation whereas the Th2 line required the addition of soluble anti-CD28 Ab to induce proliferation and IL-5 production, but this requirement could be overcome with high concentrations of plate-bound monomer alone. IL-2 production was dependent on costimulation in both cell lines. These findings demonstrate that upon antigenic rechallenge, Th1 and Th2 cells differ in their response to Ag-specific stimulation. Th2 cells were sensitive to the strength of signal to a greater degree than Th1 cells and required costimulation through CD28 for maximal proliferation. These distinctions between Th1 and Th2 activation are not consistent with a simple avidity model of Ag recognition and indicate both qualitative and quantitative differences in determining cell lineage commitment. PMID- 12538680 TI - TCR reserve: a novel principle of CD4 T cell activation by weak ligands. AB - Some ligand-receptor systems have a receptor reserve where a maximal response can be achieved by occupation of a fraction of available receptors. An implication of a receptor reserve is the expansion of the number of ligands for response. To determine whether T cells follow receptor reserve, we have characterized the effect of reducing TCR levels on CD4 T cell responses elicited by altered peptide ligands that vary in potency. Agonist peptide is unaffected by a 90% reduction in TCR level while proliferation to weak agonists is significantly inhibited when TCR expression is reduced by 40%. Thymocyte-negative selection similarly demonstrates a differential requirement of TCR for response to agonist, weak agonist, and partial agonist. Therefore, our data demonstrate receptor reserve as a novel principle of T cell activation in which excess TCRs expand the antigenic repertoire to include less potent ligands. PMID- 12538681 TI - Preferential escape of subdominant CD8+ T cells during negative selection results in an altered antiviral T cell hierarchy. AB - Negative selection is designed to purge the immune system of high-avidity, self reactive T cells and thereby protect the host from overt autoimmunity. In this in vivo viral infection model, we show that there is a previously unappreciated dichotomy involved in negative selection in which high-avidity CD8(+) T cells specific for a dominant epitope are eliminated, whereas T cells specific for a subdominant epitope on the same protein preferentially escape deletion. Although this resulted in significant skewing of immunodominance and a substantial depletion of the most promiscuous T cells, thymic and/or peripheral deletion of high-avidity CD8(+) T cells was not accompanied by any major change in the TCR V beta gene family usage or an absolute deletion of a single preferred complementarity-determining region 3 length polymorphism. This suggests that negative selection allows high-avidity CD8(+) T cells specific for subdominant or cryptic epitopes to persist while effectively deleting high-avidity T cells specific for dominant epitopes. By allowing the escape of subdominant T cells, this process still preserves a relatively broad peripheral TCR repertoire that can actively participate in antiviral and/or autoreactive immune responses. PMID- 12538682 TI - Role for IL-10 in suppression mediated by peptide-induced regulatory T cells in vivo. AB - Regulatory CD4(+) T cells were induced in the Tg4 TCR transgenic mouse specific for the N-terminal peptide (Ac1-9) of myelin basic protein by intranasal administration of a high-affinity MHC-binding analog (Ac1-9[4Y]). Peptide-induced tolerant cells (PItol) were anergic, failed to produce IL-2, but responded to Ag by secretion of IL-10. PItol cells were predominantly CD25(-) and CTLA-4(+) and their anergic state was reversed by addition of IL-2 in vitro. PItol cells suppressed the response of naive Tg4 cells both in vitro and in vivo. The in vitro suppression mediated by these cells was not reversed by cytokine neutralization and was cell-cell contact-dependent. However, suppression of proliferation and IL-2 production by PItol cells in vivo was abrogated by neutralization of IL-10. These results emphasize an important role for IL-10 in the function of peptide-induced regulatory T cells in vivo and highlight the caution required in extrapolating mechanisms of T regulatory cell function from in vitro studies. PMID- 12538683 TI - Critical role of MHC class I-related chain A and B expression on IFN-alpha stimulated dendritic cells in NK cell activation: impairment in chronic hepatitis C virus infection. AB - Dendritic cells (DCs) augment effector functions of NK cells, but the underlying mechanisms are not fully understood. Here we show in an in vitro coculture system that human monocyte-derived DCs enhance IFN-gamma production, CD69 expression, and K562 cytolytic ability of NK cells when DCs are prestimulated with various maturation stimuli such as IFN-alpha or LPS. Of interest is the finding that NK cell activation mediated by LPS-stimulated DCs was dependent on IL-12 produced in DC/NK coculture, but that IFN-alpha-stimulated DC-mediated activation was not. Alternatively, MHC class I-related chain A and B (MICA/B), ligands for NKG2D activating receptor, were found to be induced on DCs upon IFN-alpha stimulation and to be responsible for the NK activation because mAb-mediated masking of MICA/B as well as inhibition of direct cell-to-cell contact using transwell insert completely abolished DC-dependent NK cell activation by IFN-alpha. Finally, DCs recovered from chronic hepatitis C virus-infected patients showed defects in the induction of MICA/B and impaired ability to activate NK cells in response to IFN-alpha stimulation. These findings suggested that MICA/B induction on DCs may be one of the mechanisms by which IFN-alpha activates NK cells; this impairment might affect IFN-alpha responsiveness in hepatitis C virus infection. PMID- 12538684 TI - Blockade of programmed death-1 ligands on dendritic cells enhances T cell activation and cytokine production. AB - Programmed death-1 ligand (PD-L)1 and PD-L2 are ligands for programmed death-1 (PD-1), a member of the CD28/CTLA4 family expressed on activated lymphoid cells. PD-1 contains an immunoreceptor tyrosine-based inhibitory motif and mice deficient in PD-1 develop autoimmune disorders suggesting a defect in peripheral tolerance. Human PD-L1 and PD-L2 are expressed on immature dendritic cells (iDC) and mature dendritic cells (mDC), IFN-gamma-treated monocytes, and follicular dendritic cells. Using mAbs, we show that blockade of PD-L2 on dendritic cells results in enhanced T cell proliferation and cytokine production, including that of IFN-gamma and IL-10, while blockade of PD-L1 results in similar, more modest, effects. Blockade of both PD-L1 and PD-L2 showed an additive effect. Both whole mAb and Fab enhanced T cell activation, showing that PD-L1 and PD-L2 function to inhibit T cell activation. Enhancement of T cell activation was most pronounced with weak APC, such as iDCs and IL-10-pretreated mDCs, and less pronounced with strong APC such as mDCs. These data are consistent with the hypothesis that iDC have a balance of stimulatory vs inhibitory molecules that favors inhibition, and indicate that PD-L1 and PD-L2 contribute to the poor stimulatory capacity of iDC. PD-L1 expression differs from PD-L2 in that PD-L1 is expressed on activated T cells, placental trophoblasts, myocardial endothelium, and cortical thymic epithelial cells. In contrast, PD-L2 is expressed on placental endothelium and medullary thymic epithelial cells. PD-L1 is also highly expressed on most carcinomas but minimally expressed on adjacent normal tissue suggesting a role in attenuating antitumor immune responses. PMID- 12538685 TI - Enhanced differentiation of splenic plasma cells but diminished long-lived high affinity bone marrow plasma cells in aged mice. AB - In the present work, we have dissected the mechanisms responsible for the impaired humoral responses in aging. We found that there was a substantially higher level of Ab-forming cells in the spleens of aged mice than that of young controls. However, the number of high-affinity, class-switched Ab-forming cells was severely decreased in the spleen of aged mice. The accumulation of low affinity IgM Ab-forming cells in the spleens of aged animals was not due to a deficiency in isotype switching because the number of total IgG1 splenic plasma cells was not significantly reduced. Remarkably, plasma cells of both low and high affinity were significantly diminished in the bone marrow of aged mice compared with that of young mice. The results from reconstitution experiments showed that aged bone marrow was less supportive for plasma cells derived from young splenic B cells. These findings suggest that humoral immune deficiency in aging results from at least two mechanisms: the inability to generate sufficient numbers of high-affinity Ab-forming cells, which is a result of diminished germinal center reaction, and the defective bone marrow environment that has diminished ability to support the selection and survival of long-term Ab-forming cells. PMID- 12538686 TI - In vivo blockade of macrophage migration inhibitory factor ameliorates acute experimental autoimmune encephalomyelitis by impairing the homing of encephalitogenic T cells to the central nervous system. AB - Macrophage migration inhibitory factor (MIF) is a cytokine that plays a critical role in the regulation of macrophage effector functions and T cell activation. However, its role in the pathogenesis of T cell-mediated autoimmune diseases, such as experimental autoimmune encephalomyelitis (EAE), has remained unresolved. In this study, we report that anti-MIF Ab treatment of SJL mice with acute EAE improved the disease severity and accelerated the recovery. Furthermore, the anti MIF treatment impaired the homing of neuroantigen-reactive pathogenic T cells to the CNS in a VCAM-1-dependent fashion. Interestingly, MIF blockade also decreased the clonal size of the neuroantigen-specific Th1 cells and increased their activation threshold. Taken together, the results demonstrate an important role for MIF in the pathogenesis of EAE/multiple sclerosis and suggest that MIF blockade may be a promising new strategy for the treatment of multiple sclerosis. PMID- 12538687 TI - Ly6C induces clustering of LFA-1 (CD11a/CD18) and is involved in subtype-specific adhesion of CD8 T cells. AB - Ly6C is a hemopoietic cell differentiation Ag found on a subset of CD8 T cells in the periphery. It is involved in target cell killing by CTLs, augments TCR mediated activation of IL-2 and IFN-gamma production in CD8 T cells, and regulates CD8 T cell homing in vivo. In this study, we show that cross-linking of Ly6C causes clustering of LFA-1 (CD11a/CD18) on the surface of CD8 T cells via a mechanism dependent on reorganization of actin cytoskeleton and intracellular protease, calpain, but not the phosphatidylinositol 3-kinase pathway. In the capillary flow-adhesion assay, Ly6C cross-linking significantly augments lymphocyte adhesion to endothelium, and this is inhibited by an Ab that blocks LFA-1 function. Furthermore, upon in vitro cross-linking and during in vivo homing into lymph nodes, Ly6C is transiently lost from cell surface but becomes re-expressed on lymph node-resident CD8 T cells. The abilities of Ly6C to induce LFA-1 clustering and to be re-expressed after signaling-associated down regulation may be important in regulating the homing of CD8 T cells into lymph nodes and in subsequent steps of CD8 T cell activation and effector function that again involve LFA-1. PMID- 12538688 TI - Induction of antigen-specific CTL by recombinant HIV trans-activating fusion protein-pulsed human monocyte-derived dendritic cells. AB - Several systems have been tested for introduction of Ags into human dendritic cells (DC). Most of them to date, however, are complex and possess limited efficiency. Recent advances in HIV trans-activating (TAT) fusion protein technology permit extremely high transduction efficiencies for a majority of mammalian cell types. Here we report our attempts to develop a simple, but highly efficient, protocol for loading of antigenic protein into DC using TAT fusion technology. A TAT-minigene fusion protein was generated, encoding both the HLA-A2 restricted influenza matrix protein-derived epitope (GILVFTFTL, Flu-M1) and a melanoma Ag gp100-derived modified epitope (YLEPGPVTV, G9(280)-9V). In addition, both a TAT-Her2/neu extracellular domain (ECD) fusion protein and a TAT-green fluorescence protein fusion protein were generated. Over 95% of DC stained positively for TAT-green fluorescence protein within 20 min of coculture. DC treated with TAT-minigene were efficiently recognized by both Flu-M1 and G9(280) 9V-specific T cells in cytotoxicity assays and IFN-gamma ELISPOT assays. In contrast, DC pulsed with minigene fusion protein lacking TAT were either poorly recognized or not recognized by the T cells. DC pulsed with TAT-minigene also efficiently induced Flu-M1-specific T cells from naive lymphocytes. Similarly, DC treated with TAT-Her2/neu ECD stimulated patient-derived lymphocytes that specifically recognized Her2/neu(+) ovarian and breast cancer cell lines. The CTL induced by TAT-Her2/neu ECD-pulsed DC specifically recognized the Her2/neu ECD derived immunogenic peptide E75 (KIFGSLAFL). Our data suggest that TAT fusion proteins efficiently transduce DC and induce Ag-specific T cells. This could prove to be a useful method for treatment of infectious diseases and cancer. PMID- 12538689 TI - Entry into the thymic microenvironment triggers Notch activation in the earliest migrant T cell progenitors. AB - Interactions between T cell precursors and thymic stromal cells are essential during thymocyte development. However, the role of the thymus in initial commitment of lymphoid progenitors to the T lineage remains controversial, with data providing evidence for both extra- and intrathymic commitment mechanisms. In this context, it is clear that Notch1 is an important mediator during initiation of T cell development. Here we have analyzed the mechanisms regulating Notch activation in lymphoid precursors at extrathymic sites and in the thymus, including stages representing the first wave of embryonic thymus colonization on embryonic day 12 of gestation. We show that Notch activation in migrant lymphoid precursors requires entry into the thymic microenvironment where they are exposed to Notch ligands expressed by immature thymic epithelial cells. Moreover, continued Notch signaling in such precursors requires sustained interactions with Notch ligands. Collectively, these findings suggest a role for Notch in an intrathymic mechanism of T cell lineage commitment involving sustained interactions with Notch ligand bearing thymic epithelium. PMID- 12538690 TI - Inhibitory feedback loop between tolerogenic dendritic cells and regulatory T cells in transplant tolerance. AB - An active role of T regulatory cells (Treg) and tolerogenic dendritic cells (Tol DC) is believed important for the induction and maintenance of transplantation tolerance. However, interactions between these cells remain unclear. We induced donor-specific tolerance in a fully MHC-mismatched murine model of cardiac transplantation by simultaneously targeting T cell and DC function using anti CD45RB mAb and LF 15-0195, a novel analog of the antirejection drug 15 deoxyspergualin, respectively. Increases in splenic Treg and Tol-DC were observed in tolerant recipients as assessed by an increase in CD4(+)CD25(+) T cells and DC with immature phenotype. Both these cell types exerted suppressive effects in MLR. Tol-DC purified from tolerant recipients incubated with naive T cells induced the generation/expansion of CD4(+)CD25(+) Treg. Furthermore, incubation of Treg isolated from tolerant recipients with DC progenitors resulted in the generation of DC with Tol-DC phenotype. Treg and Tol-DC generated in vitro were functional based on their suppressive activity in vitro. These results are consistent with the notion that tolerance induction is associated with a self maintaining regulatory loop in which Tol-DC induce the generation of Treg from naive T cells and Treg programs the generation of Tol-DC from DC progenitors. PMID- 12538691 TI - TGF-beta regulates airway responses via T cells. AB - Allergic asthma is characterized by airway hyperreactivity, inflammation, and a Th2-type cytokine profile favoring IgE production. Beneficial effects of TGF-beta and conflicting results regarding the role of Th1 cytokines have been reported from murine asthma models. In this study, we examined the T cell as a target cell of TGF-beta-mediated immune regulation in a mouse model of asthma. We demonstrate that impairment of TGF-beta signaling in T cells of transgenic mice expressing a dominant-negative TGF-beta type II receptor leads to a decrease in airway reactivity in a non-Ag-dependent model. Increased serum levels of IFN-gamma can be detected in these animals. In contrast, after injection of OVA adsorbed to alum and challenge with OVA aerosol, transgenic animals show an increased airway reactivity and inflammation compared with those of wild-type animals. IL-13 levels in bronchoalveolar lavage fluid and serum as well as the number of inducible NO synthase-expressing cells in lung infiltrates were increased in transgenic animals. These results demonstrate an important role for TGF-beta signaling in T cells in the regulation of airway responses and suggest that the beneficial effects observed for TGF-beta in airway hyperreactivity and inflammation may be due to its regulatory effects on T cells. PMID- 12538692 TI - TNF-alpha plus IFN-gamma induce connexin43 expression and formation of gap junctions between human monocytes/macrophages that enhance physiological responses. AB - In this work, the effects of bacterial LPS, TNF-alpha, and IFN-gamma on gap junctional communication (dye coupling) and on the expression of connexin43 (immunofluorescence, immunoblotting, and RT-PCR) in monocytes/macrophages were studied. Freshly isolated human monocytes plated at high density and treated either with LPS plus IFN-gamma or TNF-alpha plus IFN-gamma became transiently dye coupled (Lucifer yellow) within 24 h. Cells treated with LPS, TNF-alpha, or IFN gamma alone remained dye uncoupled. In dye-coupled cells, the spread of Lucifer yellow to neighboring cells was reversibly blocked with 18 alpha-glycyrrhetinic acid, a gap junction blocker, but it was unaffected by oxidized ATP or probenecid, which block ionotropic ATP-activated channels and organic anion transporters, respectively. Abs against TNF-alpha significantly reduced the LPS plus IFN-gamma-induced increase in dye coupling. In dye-coupled monocytes/macrophages, but not in control cells, both connexin43 protein and mRNA were detected, and their levels were higher in cells with an elevated incidence of dye coupling. In dye-coupled cells, the localization of connexin43 immunoreactivity was diffuse at perinuclear regions and thin cell processes. The addition of 18-alpha-glycyrrhetinic acid induced a profound reduction of monocyte/macrophage transmigration across a blood brain barrier model. It also induced a significant reduction in the secretion of metalloproteinase-2 in cells treated with TNF-alpha plus IFN-gamma. We propose that some monocyte/macrophage responses are coordinated by connexin-formed membrane channels expressed transiently at inflammatory sites in which these cells form aggregates. PMID- 12538693 TI - MHC class II-peptide complexes and APC lipid rafts accumulate at the immunological synapse. AB - Activation of CD4(+) Th cells requires their cognate interaction with APCs bearing specific relevant MHC class II-peptide complexes. This cognate interaction culminates in the formation of an immunological synapse that contains the various proteins and lipids required for efficient T cell activation. We now show that APC lipid raft membrane microdomains contain specific class II-peptide complexes and serve as platforms that deliver these raft-associated class II molecules to the immunological synapse. APC rafts are required for T cell:APC conjugate formation and T cell activation at low densities of relevant class II peptide complexes, a requirement that can be overcome at high class II-peptide density. Analysis of confocal microscopy images revealed that over time APC lipid rafts, raft-associated relevant class II-peptide complexes, and even immunologically irrelevant class II molecules accumulate at the immunological synapse. As the immunological synapse matures, relevant class II-peptide complexes are sorted to a central region of the interface, while irrelevant class II molecules are excluded from this site. We propose that T cell activation is facilitated by recruitment of MHC class II-peptide complexes to the immunological synapse by virtue of their constitutive association with lipid raft microdomains. PMID- 12538694 TI - Rules for gene usage inferred from a comparison of large-scale gene expression profiles of T and B lymphocyte development. AB - Ribonucleic acid expression profiles of seven consecutive stages of mouse thymocyte development were generated on high density oligonucleotide arrays. Previously known expression patterns of several genes were confirmed. Ten percent (1,304 of more than 13,000) of the monitored genes were found with 99% confidence to be differentially expressed across all T cell developmental stages. When compared with 1,204 genes differentially expressed in five consecutive B lineage developmental stages of bone marrow, >40% (546 genes) appeared to be shared by both lineages. However, when four pools of functionally distinct cell stages were compared between B and T cell development, DJ-rearranged precursor cells and resting immature precursor cells before and after surface Ag receptor expression shared less than 10%, mature resting lymphocytes between 15 and 20%, and only cycling precursors responding to precursor lymphocyte receptor deposition shared >50% of these differentially expressed genes. Three general rules emerge from these results: 1) proliferation of cells at comparable stages is in majority executed by the same genes; 2) intracellular signaling and intercellular communication are effected largely by different genes; and 3) most genes are not used strictly at comparable, but rather at several, stages, possibly in different functional contexts. PMID- 12538695 TI - Block in development at the pre-B-II to immature B cell stage in mice without Ig kappa and Ig lambda light chain. AB - Silencing individual C (constant region) lambda genes in a kappa(-/-) background reduces mature B cell levels, and L chain-deficient (lambda(-/-)kappa(-/-)) mice attain a complete block in B cell development at the stage when L chain rearrangement, resulting in surface IgM expression, should be completed. L chain deficiency prevents B cell receptor association, and L chain function cannot be substituted (e.g., by surrogate L chain). Nevertheless, precursor cell levels, controlled by developmental progression and checkpoint apoptosis, are maintained, and B cell development in the bone marrow is fully retained up to the immature stage. L chain deficiency allows H chain retention in the cytoplasm, but prevents H chain release from the cell, and as a result secondary lymphoid organs are B cell depleted while T cell levels remain normal. PMID- 12538696 TI - Mimicry of native peptide antigens by the corresponding retro-inverso analogs is dependent on their intrinsic structure and interaction propensities. AB - Retro-inverso (ri) analogs of model T cell and B cell epitopes were predictively designed as mimics and then assayed for activity to understand the basis of functional ri-antigenic peptide mimicry. ri versions of two MHC class I binding peptide epitopes, one from a vesicular stomatitis virus glycoprotein (VSV(p)) and another from OVA (OVAp), exhibit structural as well as functional mimicry of their native counterparts. The two ri peptides exhibit conformational plasticity and they bind to MHC class I (H-2K(b)) similar to their native counterparts both in silico and in vivo. In fact, ri-OVAp is also presented to an OVAp-specific T cell line in a mode similar to native OVAp. In contrast, the ri version of an immunodominant B cell peptide epitope from a hepatitis B virus protein, PS1, exhibits no structural or functional correlation with its native counterpart. PS1 and its ri analog do not exhibit similar conformational propensities. PS1 is less flexible relative to its ri version. These observed structure-function relationships of the ri-peptide epitopes are consistent with the differences in recognition properties between peptide-MHC vs peptide-Ab binding where, while the recognition of the epitope by MHC is pattern based, the exquisitely specific recognition of Ag by Ab arises from the high complementarity between the Ag and the binding site of the Ab. It is evident that the correlation of conformational and interaction propensities of native L-peptides and their ri counterparts depends both on their inherent structural properties and on their mode of recognition. PMID- 12538697 TI - Natural substrates and inhibitors of mannan-binding lectin-associated serine protease-1 and -2: a study on recombinant catalytic fragments. AB - Mannan-binding lectin-associated serine protease (SP) (MASP)-1 and MASP-2 are modular SP and form complexes with mannan-binding lectin, the recognition molecule of the lectin pathway of the complement system. To characterize the enzymatic properties of these proteases we expressed their catalytic region, the C-terminal three domains, in Escherichia coli. Both enzymes autoactivated and cleaved synthetic oligopeptide substrates. In a competing oligopeptide substrate library assay, MASP-1 showed extreme Arg selectivity, whereas MASP-2 exhibited a less restricted, trypsin-like specificity. The enzymatic assays with complement components showed that cleavage of intact C3 by MASP-1 and MASP-2 was detectable, but was only approximately 0.1% of the previously reported efficiency of C3bBb, the alternative pathway C3-convertase. Both enzymes cleaved C3i 10- to 20-fold faster, but still at only approximately 1% of the efficiency of MASP-2 cleavage of C2. We believe that C3 is not the natural substrate of either enzyme. MASP-2 cleaved C2 and C4 at high rates. To determine the role of the individual domains in the catalytic region of MASP-2, the second complement control protein module together with the SP module and the SP module were also expressed and characterized. We demonstrated that the SP domain alone can autoactivate and cleave C2 as efficiently as the entire catalytic region, while the second complement control protein module is necessary for efficient C4 cleavage. This behavior strongly resembles C1s. Each MASP-1 and MASP-2 fragment reacted with C1 inhibitor, which completely blocked the enzymatic action of the enzymes. Nevertheless, relative rates of reaction with alpha-2-macroglobulin and C1 inhibitor suggest that alpha-2-macroglobulin may be a significant physiological inhibitor of MASP-1. PMID- 12538698 TI - Suppressor of cytokine signaling 1 inhibits IL-10-mediated immune responses. AB - IL-10 has proved to be a key cytokine in regulating inflammatory responses by controlling the production and function of various other cytokines. The suppressor of cytokine signaling (SOCS) gene products are a family of cytoplasmic molecules that are essential mediators for negatively regulating cytokine signaling. It has been previously shown that IL-10 induced SOCS3 expression and that forced constitutive expression of SOCS3 inhibits IL-10/STAT3 activation and LPS-induced macrophage activation. In this report, we show that, in addition to SOCS3 expression, IL-10 induces SOCS1 up-regulation in all cell lines tested, including Ba/F3 pro-B cells, MC/9 mast cells, M1 leukemia cells, U3A human fibroblasts, and primary mouse CD4(+) T cells. Induction of SOCS molecules is dependent on STAT3 activation by IL-10R1. Cell lines constitutively overexpressing SOCS proteins demonstrated that SOCS1 and SOCS3, but not SOCS2, are able to partially inhibit IL-10-mediated STAT3 activation and proliferative responses. Pretreatment of M1 cells with IFN-gamma resulted in SOCS1 induction and a reduction of IL-10-mediated STAT3 activation and cell growth inhibition. IL 10-induced SOCS is associated with the inhibition of IFN-gamma signaling in various cell types, and this inhibition is independent of C-terminal serine residues of the IL-10R, previously shown to be required for other anti inflammatory responses. Thus, the present results show that both SOCS1 and SOCS3 are induced by IL-10 and may be important inhibitors of both IL-10 and IFN-gamma signaling. IL-10-induced SOCS1 may directly inhibit IL-10 IFN-gamma signaling, while inhibition of other proinflammatory cytokine responses may use additional IL-10R1-mediated mechanisms. PMID- 12538699 TI - The shift of Th1 to Th2 immunodominance associated with the chronicity of Mycobacterium bovis bacille Calmette-Guerin infection does not affect the memory response. AB - In the present study we investigated the shaping and evolution of the immunodominance of the T cell response during a chronic mycobacterial infection. Using a recombinant bacille Calmette-Guerin expressing a reporter Ag, the Escherichia coli MalE protein, we analyzed the peptide specificity and the cytokine profile of the T cell response to the reporter Ag by ELISPOT. During the early steps of infection, the T cell response was focused on two dominant MalE epitopes and was characterized by a pure IFN-gamma response. Then, in the course of infection the initial IFN-gamma response to these two epitopes shifted to a mixed IFN-gamma/IL-4 response. At the same time, the peptide specificity of the T cell response was broadened to two additional MalE epitopes characterized by a unique IL-4 response resulting in the establishment of a dominant IL-4 response to the MalE protein at 16 wk postinfection. However, this phenomenon did not impair the outcome of a predominant IFN-gamma response upon subsequent MalE recall in vivo performed in the presence of CFA, a Th1-driving adjuvant. These results indicate that the Th2 nature of the immune response established during a chronic infection, which most likely reflects regulatory mechanisms to allow the return to T cell homeostasis, does not shape the Th1/Th2 nature of the memory response. PMID- 12538700 TI - Lipopolysaccharide (LPS)-binding protein mediates LPS detoxification by chylomicrons. AB - Chylomicrons have been shown to protect against endotoxin-induced lethality. LPS binding protein (LBP) is involved in the inactivation of bacterial toxin by lipoproteins. The current study examined the interaction among LBP, chylomicrons, and bacterial toxin. LBP was demonstrated to associate with chylomicrons and enhance the amount of LPS binding to chylomicrons in a dose-dependent fashion. In addition, LBP accelerated LPS binding to chylomicrons. This LBP-induced interaction of LPS with chylomicrons prevented endotoxin toxicity, as demonstrated by reduced cytokine secretion by PBMC. When postprandial circulating concentrations of chylomicrons were compared with circulating levels of low density lipoprotein, very low density lipoprotein, and high density lipoprotein, chylomicrons exceeded the other lipoproteins in LPS-inactivating capacity. Furthermore, highly purified lipoteichoic acid, an immunostimulatory component of Gram-positive bacteria, was detoxified by incubation with LBP and chylomicrons. In conclusion, our results indicate that LBP associates with chylomicrons and enables chylomicrons to rapidly bind bacterial toxin, thereby preventing cell activation. Besides a role in the detoxification of bacterial toxin present in the circulation, we believe that LBP-chylomicron complexes may be part of a local defense mechanism of the intestine against translocated bacterial toxin. PMID- 12538701 TI - Human intestinal epithelial cells are broadly unresponsive to Toll-like receptor 2-dependent bacterial ligands: implications for host-microbial interactions in the gut. AB - Intestinal epithelial cells (IEC) interact with a high density of Gram-positive bacteria and are active participants in mucosal immune responses. Recognition of Gram-positive organisms by Toll-like receptor (TLR)2 induces proinflammatory gene expression by diverse cells. We hypothesized that IEC are unresponsive to Gram positive pathogen-associated molecular patterns and sought to characterize the functional responses of IEC to TLR2-specific ligands. Human colonic epithelial cells isolated by laser capture microscopy and IEC lines (Caco-2, T84, HT-29) were analyzed for expression of TLR2, TLR6, TLR1, and Toll inhibitory protein (Tollip) mRNA by RT-PCR and quantitative real-time PCR. Response to Gram-positive bacterial ligands was measured by NF-kappa B reporter gene activation and IL-8 secretion. TLR2 protein expression was analyzed by immunofluorescence and flow cytometry. Colonic epithelial cells and lamina propria cells from both uninflamed and inflamed tissue demonstrate low expression of TLR2 mRNA compared with THP-1 monocytes. IECs were unresponsive to TLR2 ligands including the staphylococcal derived Ags phenol soluble modulin, peptidoglycan, and lipotechoic acid and the mycobacterial-derived Ag soluble tuberculosis factor. Transgenic expression of TLR2 and TLR6 restored responsiveness to phenol soluble modulin and peptidoglycan in IEC. In addition to low levels of TLR2 protein expression, IEC also express high levels of the inhibitory molecule Tollip. We conclude that IEC are broadly unresponsive to TLR2 ligands secondary to deficient expression of TLR2 and TLR6. The relative absence of TLR2 protein expression by IEC and high level of Tollip expression may be important in preventing chronic proinflammatory cytokine secretion in response to commensal Gram-positive bacteria in the gut. PMID- 12538702 TI - A simian replication-defective adenoviral recombinant vaccine to HIV-1 gag. AB - In animal models, E1-deleted human adenoviral recombinants of the serotype 5 (AdHu5) have shown high efficacy as vaccine carriers for different Ags including those of HIV-1. Humans are infected by common serotypes of human adenovirus such as AdHu5 early in life and a significant percentage has high levels of neutralizing Abs to these serotypes, which will very likely impair the efficacy of recombinant vaccines based on the homologous virus. To circumvent this problem, a novel replication-defective adenoviral vaccine carrier based on an E1 deleted recombinant of the chimpanzee adenovirus 68 (AdC68) was developed. An AdC68 construct expressing a codon-optimized, truncated form of gag of HIV-1 induces CD8(+) T cells to gag in mice which at the height of the immune response encompass nearly 20% of the entire splenic CD8(+) T cell population. The vaccine induced immune response provides protection to challenge with a vaccinia gag recombinant virus. Induction of transgene-specific CD8(+) T cells and protection against viral challenge elicited by the AdC68 vaccines is not strongly inhibited in animals preimmune to AdHu5 virus. However, the response elicited by the AdHu5 vaccine is greatly attenuated in AdHu5 preimmune animals. PMID- 12538703 TI - Nonspecific recruitment of memory CD8+ T cells to the lung airways during respiratory virus infections. AB - Previous studies have shown that heterologous viral infections have a significant impact on pre-existing memory T cell populations in secondary lymphoid organs through a combination of cross-reactive and bystander effects. However, the impact of heterologous viral infections on effector/memory T cells in peripheral sites is not well understood. In this study, we have analyzed the impact of a heterologous influenza virus infection on Sendai virus-specific CD8(+) effector/memory cells present in the lung airways. The data show a transient increase in the numbers of Sendai virus nucleoprotein 324-332/K(b)-specific CD8(+) memory T cells in the airways of the influenza-infected mice peaking around day 4 postinfection. Intratracheal transfer studies and 5-bromo-2' deoxyuridine incorporation demonstrate that this increase is due to the recruitment of resting memory cells into the airways. In addition, the data show that these immigrating memory cells are phenotypically distinct from the resident memory T cells of the lung airways. A similar influx of nonproliferating Sendai virus nucleoprotein 324-332/K(b)-specific CD8(+) memory T cells is also induced by a secondary (homologous) infection with Sendai virus. Together, these data suggest that inflammation can accelerate memory T cell migration to nonlymphoid tissues and is a part of the normal recall response during respiratory infections. PMID- 12538704 TI - Impaired clearance of herpes simplex virus type 1 from mice lacking CD1d or NKT cells expressing the semivariant V alpha 14-J alpha 281 TCR. AB - Ag-presenting molecule CD1 and CD1-restricted NKT cells are known to contribute to defense against a range of infectious pathogens, including some viruses. CD1 restricted NKT cells, a distinct subpopulation of T cells, have striking and rapid effector functions that contribute to host defense, including rapid production of IFN-gamma and IL-4, and activation of NK cells. Consideration of the important contributions of innate and adaptive immunity to clearance of HSV prompted us to investigate the role of CD1 and of NKT cells expressing the V alpha 14-J alpha 281 TCR in the pathogenesis of HSV infection. To address this issue, we compared infection in wild-type mice with that in CD1 gene knockout (GKO) and J alpha 281 GKO mice. In this study, we report impaired clearance of virus and viral Ags, and more florid acute infection in mice lacking CD1 (and by inference, CD1-restricted T cells), in comparison with parental C57BL6 mice. In J alpha 281 GKO mice there was also impairment of virus clearance, resembling that seen in CD1 GKO mice. These results imply roles for the V alpha 14-J alpha 281 subset of NKT cells and for CD1d in control of HSV infection. PMID- 12538705 TI - A soluble chemokine-binding protein from vaccinia virus reduces virus virulence and the inflammatory response to infection. AB - Many poxviruses express a secreted protein that binds CC chemokines with high affinity and has been called viral CC chemokine inhibitor (vCCI). This protein is unrelated to any known cellular protein, yet can compete with host cellular CC chemokine receptors to modulate host inflammatory and immune responses. Although several strains of vaccinia virus (VV) express a vCCI, the best characterized VV strains Western Reserve and Copenhagen do not. In this study, we have expressed the vCCI from VV strain Lister in a recombinant Western Reserve virus (v Delta B8R-35K) and characterized its binding properties in vitro and its effect on virulence in vivo relative to wild-type virus (v Delta B8R) or a revertant virus (v Delta B8R-R) where Lister 35-kDa had been removed. Cells infected with v Delta B8R-35K secreted a 35-kDa protein that bound the CC chemokine macrophage inflammatory protein 1 alpha. Expression of vCCI attenuated the virus in a murine intranasal model, characterized by reduced mortality and weight loss, decreased virus replication and spread, and a reduced recruitment of inflammatory cells into the lungs of VV-infected mice. The CC chemokines macrophage-inflammatory protein 1 alpha, eotaxin, and macrophage chemotactic protein 1 were detected in bronchoalveolar lavage fluids from v Delta B8R-infected mice; however, bronchoalveolar lavage fluids from v Delta B8R-35K-infected mice had lower levels of chemokines and a reduced chemotactic activity for murine leukocytes in vitro. These observations suggest that vCCI plays an important role in regulating leukocyte trafficking to the lungs during VV infection by binding to CC chemokines and blocking their chemotactic activities. PMID- 12538706 TI - A specific role for B cells in the generation of CD8 T cell memory by recombinant Listeria monocytogenes. AB - In this study, we investigated whether B cells play a role in the induction and maintenance of CD8 T cell memory after immunization with an intracellular bacterium, Listeria monocytogenes. Our results show that B cells play a minimal role in the initial activation and Ag-driven expansion of CD8 T lymphocytes. However, absence of B cells results in increased death of activated CD8 T cells during the contraction phase, leading to a lower level of Ag-specific CD8 T cell memory. Once memory is established, B cells are no longer required for the long term maintenance and rapid recall response of memory CD8 T cells. Increased contraction of Ag-specific CD8 T cells in B cell-deficient mice is not due to impaired CD4 T cell responses since priming of epitope-specific CD4 T cell responses is normal in B cell-deficient mice following L. monocytogenes infection. Furthermore, no exaggerated contraction of Ag-specific CD8 T cells is evident in CD4 knockout mice. Thus, B cells play a specific role in modulating the contraction of CD8 T cell responses following immunization. Elucidation of factors that regulate the death phase may allow us to manipulate this process to increase the level of immunological memory and thus, vaccine efficacy. PMID- 12538707 TI - CCL28 has dual roles in mucosal immunity as a chemokine with broad-spectrum antimicrobial activity. AB - CCL28 is a CC chemokine signaling via CCR10 and CCR3 that is selectively expressed in certain mucosal tissues such as exocrine glands, trachea, and colon. Notably, these tissues commonly secrete low-salt fluids. RT-PCR analysis demonstrated that salivary glands expressed CCL28 mRNA at the highest levels among various mouse tissues. Single cells prepared from mouse parotid glands indeed contained a major fraction of CD3(-)B220(low) cells that expressed CCR10 at high levels and CCR3 at low levels and responded to CCL28 in chemotaxis assays. Morphologically, these cells are typical plasma cells. By immunohistochemistry, acinar epithelial cells in human and mouse salivary glands were strongly positive for CCL28. Furthermore, human saliva and milk were found to contain CCL28 at high concentrations. Moreover, the C terminus of human CCL28 has a significant sequence similarity to histatin-5, a histidine-rich candidacidal peptide in human saliva. Subsequently, we demonstrated that human and mouse CCL28 had a potent antimicrobial activity against Candida albicans, Gram-negative bacteria, and Gram-positive bacteria. The C-terminal 28-aa peptide of human CCL28 also displayed a selective candidacidal activity. In contrast, CCL27, which is most similar to CCL28 and shares CCR10, showed no such potent antimicrobial activity. Like most other antimicrobial peptides, CCL28 exerted its antimicrobial activity in low-salt conditions and rapidly induced membrane permeability in target microbes. Collectively, CCL28 may play dual roles in mucosal immunity as a chemoattractant for cells expressing CCR10 and/or CCR3 such as plasma cells and also as a broad-spectrum antimicrobial protein secreted into low-salt body fluids. PMID- 12538708 TI - Differential expression of the murine mannose-binding lectins A and C in lymphoid and nonlymphoid organs and tissues. AB - Mannose-binding lectin (MBL), a member of the collectin family, binds to carbohydrate structures on the surfaces of micro-organisms and may serve as a recognition molecule of the lectin pathway of complement activation. In rodents two forms, MBL-A and MBL-C, were described and shown to be products of two related, but uncoupled, genes. The liver is the main source of MBL biosynthesis. For rat MBL-A, expression has also been described in the kidney. Here we report that the two forms of murine MBL are differentially expressed in a number of nonhepatic tissues. Real-time RT-PCR revealed that the liver is the major site of expression for both MBL genes. Lower copy numbers were found in kidney, brain, spleen, and muscle. In testis, only the MBL-A gene is expressed, whereas MBL-C is exclusively expressed in small intestine. Using in situ hybridization and immunohistochemistry, we demonstrate that both MBLs are synthesized by hepatocytes and show MBL expression in cells of the monocyte/macrophage lineage. In the kidney MBL-A, but not MBL-C, was found to be synthesized. Vice versa, only MBL-C biosynthesis was detected in endothelial cells of the small intestine. The latter finding may support the view that MBL-C, as part of the innate immune system, may be a counterpart of secretory IgA of the acquired immune system in preventing, for example, microbial invasion and colonization. Our findings demonstrate that MBL-A and MBL-C are differentially expressed, implying distinct biological roles for both recognition molecules of the murine lectin pathway of complement. PMID- 12538709 TI - The long pentraxin PTX3 is synthesized in IgA glomerulonephritis and activates mesangial cells. AB - The long pentraxin PTX3 has been recently involved in amplification of the inflammatory reactions and regulation of innate immunity. In the present study we evaluated the expression and role of PTX3 in glomerular inflammation. PTX3 expression was investigated in the IgA, type I membranoproliferative, and diffuse proliferative lupus glomerulonephritis, which are characterized by inflammatory and proliferative lesions mainly driven by resident mesangial cells, and in the membranous glomerulonephritis and the focal segmental glomerular sclerosis, where signs of glomerular inflammation are usually absent. We found an intense staining for PTX3 in the expanded mesangial areas of renal biopsies obtained from patients with IgA glomerulonephritis. The pattern of staining was on glomerular mesangial and endothelial cells. Scattered PTX3-positive cells were also detected in glomeruli of type I membranoproliferative glomerulonephritis. The concomitant expression of CD14 suggests an inflammatory origin of these cells. Normal renal tissue and biopsies from patients with the other glomerular nephropathies studied were mainly negative for PTX3 expression in glomeruli. However, PTX3-positive cells were detected in the interstitium of nephropathies showing inflammatory interstitial injury. In vitro, cultured human mesangial cells synthesized PTX3 when stimulated with TNF-alpha and IgA and exhibited specific binding for recombinant PTX3. Moreover, stimulation with exogenous PTX3 promoted mesangial cell contraction and synthesis of the proinflammatory lipid mediator platelet activating factor. In conclusion, we provide the first evidence that mesangial cells may both produce and be a target for PTX3. The detection of this long pentraxin in the renal tissue of patients with glomerulonephritis suggests its potential role in the modulation of glomerular and tubular injury. PMID- 12538710 TI - MHC class II-associated invariant chain isoforms regulate pulmonary immune responses. AB - Asthma, a chronic inflammatory disease of the lung, is characterized by reversible airway obstruction and airway hyperresponsiveness (AHR), and is associated with increased production of IgE and Th2-type cytokines (IL-4, IL-5, and IL-13). Development of inflammation within the asthmatic lung depends on MHC class II-restricted Ag presentation, leading to stimulation of CD4(+) T cells and cytokine generation. Conventional MHC class II pathways require both MHC associated invariant chain (Ii) and HLA-DM (H2-M in mice) chaperone activities, but alternative modes of Ag presentation may also promote in vivo immunity. In this study, we demonstrate that Ii(-/-) and H2-M(-/-) mice fail to develop lung inflammation or AHR following sensitization and challenge with OVA in a mouse model of allergic inflammation. To assess potentially distinct contributions by Ii chain isoforms to lung immunity, we also compared allergen-induced lung inflammation, eosinophilia, IgE production, and AHR in mice genetically altered to express either p31 Ii or p41 Ii isoform alone. Sole expression of either Ii isoform alone facilitates development of allergen-induced lung inflammation and eosinophilia. However, animals expressing only the p31 Ii isoform exhibit abrogated IgE and AHR responses as compared with p41 Ii mice in this model of allergen-induced lung inflammation, suggesting that realization of complete immunity within the lung requires expression of p41 Ii. These findings reveal a crucial role of Ii and H2-M in controlling the immune response within the lung, and suggest that p31 Ii and p41 Ii manifest nonredundant roles in development of immunity. PMID- 12538711 TI - Cytochrome P4502D6(193-212): a new immunodominant epitope and target of virus/self cross-reactivity in liver kidney microsomal autoantibody type 1 positive liver disease. AB - Cytochrome P4502D6 (CYP2D6), target of liver kidney microsomal autoantibody type 1 (LKM1), characterizes autoimmune hepatitis type 2 (AIH2) but is also found in patients with chronic hepatitis C virus (HCV) infection. To provide a complete linear epitope B cell map of CYP2D6, we tested peptides spanning the entire sequence of CYP2D6. In addition to confirming previously described antigenic sites, we identified four new epitopes (193-212, 238-257, 268-287, and 478-497). CYP2D6(193-212) is immunodominant and was the target of 12 of 13 (93%) patients with AIH2 and 5 of 10 (50%) HCV/LKM1-positive patients. Because LKM1 is present in both AIH2 and a viral infection, we tested whether Abs to CYP2D6(193-212) arise through cross-reactive immunity between virus and self. We identified a hexameric sequence "RLLDLA" sharing 5 of 6 aa with "RLLDLS" of HCV(2985-2990) and all 6 aa with CMV(130-135). Of 17 CYP2D6(193-212)-reactive sera, 11 (7 AIH and 4 HCV) reacted by ELISA with the HCV homologue, 8 (5 AIH and 3 HCV) with the CMV homologue, and 8 (5 AIH and 3 HCV) showed double reactivity. Autoantibody binding to CYP2D6(193-212) was inhibited by preincubation with HCV(2977-2996) or CMV(121 140). Recombinant HCV-nonstructural protein 5 and CMV-UL98 proteins also inhibited Ab binding to CYP2D6(193-212). Affinity-purified CYP2D6(193-212) specific Ab inhibited the metabolic activity of CYP2D6. The demonstrated similarity and cross-reactivity between CYP2D6(193-212) and two unrelated viruses suggests that multiple exposure to viruses mimicking self may represent an important pathway to the development of autoimmunity. PMID- 12538712 TI - CD4+ Th2 cell recognition of HLA-DR-restricted epitopes derived from CAMEL: a tumor antigen translated in an alternative open reading frame. AB - Tumor Ag NY-ESO-1 is an attractive target for immunotherapy of cancer, since both CD8(+) CTL and CD4(+) Th cells against NY-ESO-1 have been described. Moreover, NY ESO-1 as well as the highly homologous tumor Ag LAGE-1 are broadly expressed in various tumor types. Interestingly, the NY-ESO-1 and LAGE-1 genes also encode for proteins translated in an alternative open reading frame. These alternatively translated NY-ESO-ORF2 and CAMEL proteins, derived from the NY-ESO-1 and LAGE-1 genes, respectively, have been demonstrated to be immunogenic, since CTL specific for these proteins have been isolated from melanoma patients. In this study a panel of advanced melanoma patients was screened for the presence of Th cells specific for the alternatively translated tumor Ags NY-ESO-ORF2 and CAMEL. PBMC of melanoma patients were stimulated for 4 days with mixes of overlapping peptides covering the entire NY-ESO-ORF2 and CAMEL protein sequences and were tested for the release of type 1 (IFN-gamma) and type 2 (IL-13) cytokines in ELISPOT assays. In three of 15 patients, T cells specific for two CAMEL peptides (CAMEL(71-92) and CAMEL(81-102)) could be detected. From one of these patients, CD4(+) T cell clones specific for CAMEL(81-102) could be generated. These clones recognized a naturally processed epitope presented in both HLA-DR11 and HLA-DR12 and produced high levels of IL-4, IL-5, and IL-13. In conclusion, this study shows the presence of Th cells specific for the alternatively translated tumor Ag CAMEL in melanoma patients and is the first report that describes the isolation of tumor Ag-specific CD4(+) Th 2 clones. PMID- 12538713 TI - Induction of antigen-specific CTL responses using antigens conjugated to short peptide vectors. AB - Linear peptides (SynB vectors) with specific sequence motifs have been identified that are capable of enhancing the transport of a wide range of molecules into cells. These peptide vectors have been used to deliver exogenous peptides and protein Ags across the cell membrane and into the cytoplasm of cells. Specifically, in vitro analysis indicated that these SynB peptides enhanced the uptake of two 9-mer peptide Ags, NP(147-155) and Mtb(250-258) (T cell epitopes of influenza nucleoprotein and Mycobacterium tuberculosis, respectively) and the M. tuberculosis Ag Mtb8.4 protein, into K562 cells when covalently linked to the respective Ags. Furthermore, selected SynB vectors, when conjugated to these same Ags and used as immunogens, resulted in considerably enhanced Ag-specific CTL responses. Several SynB vectors were tested and resulted in varying levels of cellular uptake. The efficiency of uptake correlated with the ability of the SynB construct to deliver each epitope in vivo and induce specific CTL responses in mice. These data suggest that peptide vectors, such as SynB that transport target Ags across the cell membrane in a highly efficient manner, have significant potential for vaccine delivery. PMID- 12538714 TI - Bordetella pertussis infection in 2-month-old infants promotes type 1 T cell responses. AB - Neonatal immaturity of the immune system is currently believed to generally limit the induction of immune responses to vaccine Ags and to skew them toward type 2 responses. We demonstrated here that Bordetella pertussis infection in very young infants (median, 2 mo old) as well as the first administration of whole-cell pertussis vaccine induces B. pertussis Ag-specific IFN-gamma secretion by the PBMC of these infants. IFN-gamma was secreted by both CD4(+) and CD8(+) T lymphocytes, and the levels of Ag-induced IFN-gamma secretion did not correlate with the age of the infants. Appearance of the specific Th-1 cell-mediated immunity was accompanied by a general shift of the cytokine secretion profile of these infants toward a stronger Th1 profile, as evidenced by the response to a polyclonal stimulation. We conclude that the immune system of 2-mo-old infants is developmentally mature enough to develop Th1 responses in vivo upon infection by B. pertussis or vaccination with whole-cell pertussis vaccines. PMID- 12538715 TI - CD28/B7 regulation of anti-CD3-mediated immunosuppression in vivo. AB - FcR-binding "classical" anti-CD3 mAb is a potent immunosuppressive drug that alters CD4(+) and CD8(+) T cell function in vivo via anergy induction and programmed cell death (PCD). Anti-CD3-mediated PCD was Fas independent but was mediated by the mitochondria-initiated apoptosis that was abrogated in Bcl-x(L) transgenic T cells. The PCD was more pronounced in CD28-deficient mice consistent with defective Bcl-x(L) up-regulation. Residual T cells isolated from anti-CD3 treated wild-type, CD28(-/-), and Bcl-x(L)-transgenic mice were hyporesponsive. The hyporesponsiveness was more pronounced in CD28(-/-) and wild-type mice treated with anti-B7-2, suggesting that CD28 interaction with B7-2 regulates T cell responsiveness in anti-CD3-treated animals. Finally, anti-CD3 treatment led to indefinite cardiac allograft survival in wild-type but not Bcl-x(L) animals. Together these results implicate CD28/B7 signaling in the regulation of both anti CD3-induced T cell depletion and hyporesponsiveness in vivo, but T cell depletion, not hyporesponsiveness, appears to be critical for anti-CD3 mAb mediated long-term immune regulation. PMID- 12538716 TI - Lack of a functional alternative complement pathway ameliorates ischemic acute renal failure in mice. AB - Ischemia/reperfusion (I/R) injury of the kidney is a common cause of acute renal failure (ARF) and is associated with high morbidity and mortality in the intensive care unit. The mechanisms underlying I/R injury are complex. Studies have shown that complement activation contributes to the pathogenesis of I/R injury in the kidney, but the exact mechanisms of complement activation have not been defined. We hypothesized that complement activation in this setting occurs via the alternative pathway and that mice deficient in complement factor B, an essential component of the alternative pathway, would be protected from ischemic ARF. Wild-type mice suffered from a decline in renal function and had significant tubular injury, particularly in the outer medulla, after I/R. We found that factor B-deficient mice (fB(-/-)) developed substantially less functional and morphologic renal injury after I/R. Furthermore, control wild-type mice had an increase in tubulointerstitial complement C3 deposition and neutrophil infiltration in the outer medulla after I/R, whereas fB(-/-) mice demonstrated virtually no C3 deposition or neutrophil infiltration. Our results demonstrate that complement activation in the kidney after I/R occurs exclusively via the alternative pathway, and that selective inhibition of this pathway provides protection to the kidneys from ischemic ARF. PMID- 12538717 TI - A novel anti-inflammatory role for simvastatin in inflammatory arthritis. AB - 3-Hydroxy-3-methylglutaryl-CoA reductase inhibitors (statins) exert favorable effects on lipoprotein metabolism, but may also possess anti-inflammatory properties. Therefore, we explored the activities of simvastatin, a lipophilic statin, in a Th1-driven model of murine inflammatory arthritis. We report in this study that simvastatin markedly inhibited not only developing but also clinically evident collagen-induced arthritis in doses that were unable to significantly alter cholesterol concentrations in vivo. Ex vivo analysis demonstrated significant suppression of collagen-specific Th1 humoral and cellular immune responses. Moreover, simvastatin reduced anti-CD3/anti-CD28 proliferation and IFN gamma release from mononuclear cells derived from peripheral blood and synovial fluid. Proinflammatory cytokine production in vitro by T cell contact-activated macrophages was suppressed by simvastatin, suggesting that such observations have direct clinical relevance. These data clearly illustrate the therapeutic potential of statin-sensitive pathways in inflammatory arthritis. PMID- 12538718 TI - The in vitro and in vivo effects of anti-galactose antibodies on endothelial cell activation and xenograft rejection. AB - We have previously produced a series of antigalactose (anti-Gal) hybridomas and characterized their heavy chain gene usage. Here we have quantified the affinity of these Abs for the alpha-Gal epitope and characterized their in vitro effects on endothelial cell activation and apoptosis. We report that anti-Gal mAbs derived from Gal(-/-) mice show a range of affinity for the alpha-Gal epitope, and that affinity was generally increased as the V(H) gene usage transitioned from germline sequences to sequences exhibiting somatic maturation. Despite an 85 fold range in affinity, all the anti-Gal mAbs examined induced alpha-Gal-specific endothelial cell activation, and after prolonged exposure induced endothelial cell apoptosis in a complement-independent manner. Only murine anti-Gal mAbs of the IgM or IgG3 subclass, but not IgG1, were effective at initiating complement dependent cell lysis. Using a novel rat to mouse xenograft model, we examined the in vivo ability of these mAbs to induce xenograft rejection and characterized the rejection using histology and immunohistochemistry. Infusion of complement-fixing IgG3 mAbs resulted in either hyperacute rejection or acute vascular rejection of the xenograft. Surprisingly, infusion of an equal amount of a high affinity anti Gal IgG1 mAb, that fixed complement poorly also induced a rapid xenograft rejection, which we have labeled very acute rejection. These studies emphasize the importance of in vivo assays, in addition to in vitro assays, in understanding the role of anti-Gal IgG-mediated tissue injury and xenograft rejection. PMID- 12538719 TI - Increased bone mass is a part of the generalized lymphoproliferative disorder phenotype in the mouse. AB - We investigated the bone phenotype of mice with generalized lymphoproliferative disorder (gld) due to a defect in the Fas ligand-mediated apoptotic pathway. C57BL/6-gld mice had greater whole body bone mineral density and greater trabecular bone volume than their wild-type controls. gld mice lost 5-fold less trabecular bone and had less osteoclasts on bone surfaces after ovariectomy induced bone resorption. They also formed more bone in a model of osteogenic regeneration after bone marrow ablation, had less osteoclasts on bone surfaces and less apoptotic osteoblasts. gld and wild-type mice had similar numbers of osteoclasts in bone marrow cultures, but marrow stromal fibroblasts from gld mice formed more alkaline phosphatase-positive colonies. Bone diaphyseal shafts and bone marrow stromal fibroblasts produced more osteoprotegerin mRNA and protein than wild-type mice. These findings provide evidence that the disturbance of the bone system is a part of generalized lymphoproliferative syndrome and indicates the possible role of osteoprotegerin as a regulatory link between the bone and immune system. PMID- 12538720 TI - Oral feeding with ethinyl estradiol suppresses and treats experimental autoimmune encephalomyelitis in SJL mice and inhibits the recruitment of inflammatory cells into the central nervous system. AB - There is much interest in the possible ameliorating effects of estrogen on various autoimmune diseases. We previously established the protective effects of 17 beta-estradiol (E2) on experimental autoimmune encephalomyelitis (EAE). In the current study we investigated the effectiveness of oral treatment with ethinyl estradiol (EE) on EAE and the mechanisms involved. Ethinyl estradiol is a semisynthetic estrogen compound found in birth control pills, and its chemical structure allows this compound to retain activity when given orally. We found that oral EE, like E2, drastically suppressed EAE induced by proteolipid protein 139-151 peptide when given at initiation of EAE. However, unlike E2, EE reduced clinical severity when given after the onset of clinical signs. Treatment with EE significantly decreased the secretion of proinflammatory cytokines (IFN-gamma, TNF-alpha, and IL-6) by activated T cells as well as the expression of a key matrix metalloproteinase, disease-mediating chemokines/receptors, and IgG2a levels, but increased the expression of TGF-beta 3 in the CNS. The absence of infiltrating lymphocytes together with the suppression of cytokines, matrix metalloproteinase, and chemokines/receptors suggests that EE, like E2, protects mice from EAE by inhibiting the recruitment of T cells and macrophages into the CNS. These results suggest that oral ethinyl estradiol might be a successful candidate as therapy for multiple sclerosis. PMID- 12538721 TI - Peptide nucleic acid antisense prolongs skin allograft survival by means of blockade of CXCR3 expression directing T cells into graft. AB - CXCR3, predominantly expressed on memory/activated T cells, is a receptor for both IFN-gamma-inducible protein 10/CXC chemokine ligand (CXCL)10 and monokine induced by IFN-gamma/CXCL9. It was reported that CXC chemokines IFN-gamma inducible protein 10/CXCL10 and monokine induced by IFN-gamma/CXCL9 play a critical role in the allograft rejection. We report that CXCR3 is a dominant factor directing T cells into mouse skin allograft, and that peptide nucleic acid (PNA) CXCR3 antisense significantly prolongs skin allograft survival by means of blockade of CXCR3 expression directing T cells into allografts in mice. We found that CXCR3 is highly up-regulated in spleen T cells and allografts from BALB/c recipients by day 7 of receiving transplantation, whereas CCR5 expression is moderately increased. We designed PNA CCR5 and PNA CXCR3 antisenses, and i.v. treated mice that received skin allograft transplantations. The PNA CXCR3 at a dosage of 10 mg/kg/day significantly prolonged mouse skin allograft survival (17.1 +/- 2.4 days) compared with physiological saline treatment (7.5 +/- 0.7 days), whereas PNA CCR5 (10 mg/kg/day) marginally prolonged skin allograft survival (10.7 +/- 1.1 days). The mechanism of prolongation of skin allograft survival is that PNA CXCR3 directly blocks the CXCR3 expression in T cells, which is responsible for directing T cells into skin allograft to induce acute rejection, without interfering with other functions of the T cells. These results were obtained at mRNA and protein levels by flow cytometry and real-time quantitative RT-PCR technique, and confirmed by chemotaxis, Northern and Western blot assays, and histological evaluation of skin grafts. The present study indicates the therapeutic potential of PNA CXCR3 to prevent acute transplantation rejection. PMID- 12538722 TI - Induction of apoptosis by HIV-1-infected monocytic cells. AB - We have previously described a soluble 6000-Da peptide produced by an HIV-1 infected human macrophage cell line, clone 43(HIV), which induces apoptosis in T and B cells. We have identified this factor as the novel cDNA clone FL14676485 that encodes for the human hypothetical protein, FLJ21908. The FL14676485 cDNA clone was isolated from a 43(HIV) lambda ZAP Escherichia coli expression library and screened with a panel of rabbit and mouse anti-apoptotic Abs. We transfected the FL14676485 clone into Bosc cells and non-HIV-1-infected 43 cells. Western blot analysis of lysates from the FL14676485-transfected 43 cells and Bosc cells using anti-proapoptotic factor Abs revealed a protein with a molecular mass of 66 kDa corresponding to the size of the full-length gene product of the FL14676485 clone, while Western blot of the supernatant demonstrated a doublet of 46-kDa and 6000-Da peptide that corresponds to our previously described proapoptotic factor. Primary HIV-1(BaL)-infected monocytes also produce the FLJ21908 protein. Supernatants from these transfected cells induced apoptosis in PBMC, CD4(+), and CD8(+) T and B cells similar to the activity of our previously described proapoptotic factor. PCR analysis of 43 cells and 43(HIV) cells revealed a base pair fragment of 420 bp corresponding to the FL14676485 gene product in 43(HIV) cells, but not in 43 cells. The FLJ21908 protein induces apoptosis through activation of caspase-9 and caspase-3. We have further demonstrated that the FLJ21908 protein has apoptotic activity in the SH-SY5Y neuronal cell line and can be detected in brain and lymph tissue from HIV-1-infected patients who have AIDS dementia. The FLJ21908 protein may contribute to the apoptosis and dementia observed in AIDS patients. PMID- 12538723 TI - Normal tissue depresses while tumor tissue enhances human T cell responses in vivo to a novel self/tumor melanoma antigen, OA1. AB - Antitumor T cells often recognize targets that are nonmutated "self" tissue differentiation Ags, but the relative impact of Ag expression by normal and transformed tissue for a human self/tumor Ag has not been studied. To examine the influence of self-tolerance mechanisms on the function of self/tumor-specific T cell responses in humans, we sought to identify an Ag that was expressed, processed, and presented in an MHC-restricted fashion by tumor cells, but for which there was the human equivalent of a "knockout." In this study, we report the first immunological characterization of a melanoma/melanocyte differentiation Ag, called OA1, which meets these criteria. This Ag, an X chromosome-encoded melanoma/melanocyte differentiation Ag, was completely deleted in a male patient. Using a newly identified HLA-A*2402-restricted epitope (LYSACFWWL) to study T cell tolerance, we found that OA1-specific T cell reactivity was more than five SD higher in the knockout patient that in normal controls. These data provide compelling evidence for T cell tolerance to OA1 in humans. Most surprisingly, we found elevated levels of OA1-specific T cells in patients with metastatic malignant melanoma, indicating that the tumor-bearing state partially reversed tolerance observed in normal (non-"knockout") individuals. Taken together, these findings indicated that tolerance can exist for self/tumor Ags in humans, and that this tolerance could be partially abrogated by the growth of the tumor, increasing the reactivity of tumor Ag-specific T cells. Thus, the tumor-bearing state reverses, in part, the tolerance of T cells that results from the normal expression of tissue differentiation Ags. PMID- 12538724 TI - Transcutaneous immunization with cholera toxin B subunit adjuvant suppresses IgE antibody responses via selective induction of Th1 immune responses. AB - Topical application of cholera toxin (CT) onto mouse skin can induce a humoral immune response to CT as well as to coadministered Ags. In this study, we examined the nontoxic cell-binding B subunit of CT (CTB) as a potential adjuvant for cutaneous immune responses when coadministered with the prototype protein Ag, OVA. CTB applied onto skin induced serum Ab responses to itself with magnitudes comparable to those evoked by CT but was poorly efficient at promoting systemic Ab responses to coadministered OVA. However, transcutaneous immunization (TCI) with either CT or CTB and OVA led to vigorous OVA-specific T cell proliferative responses. Furthermore, CTB potentiated Th1-driven responses (IFN-gamma production) whereas CT induced both Th1 and Th2 cytokine production. Coadministration of the toxic subunit CTA, together with CTB and OVA Ag, led to enhanced Th1 and Th2 responses. Moreover, whereas TCI with CT enhanced serum IgE responses to coadministered OVA, CTB suppressed these responses. TCI with either CT or CTB led to an increased accumulation of dendritic cells in the exposed epidermis and the underlying dermis. Thus, in contrast to CT, CTB appears to behave very differently when given by the transcutaneous as opposed to a mucosal route and the results suggest that the adjuvanticity of CT on Th1- and Th2 dependent responses induced by TCI involves two distinct moieties, the B and the A subunits, respectively. PMID- 12538725 TI - SF20/IL-25, a novel bone marrow stroma-derived growth factor that binds to mouse thymic shared antigen-1 and supports lymphoid cell proliferation. PMID- 12538726 TI - N-Glycosylation at two sites critically alters thiazide binding and activity of the rat thiazide-sensitive Na(+):Cl(-) cotransporter. AB - The rat thiazide-sensitive Na-Cl cotransporter (rNCC) is expressed in the renal distal convoluted tubule and is the site of action of an important class of antihypertensive agents, the thiazide diuretics. The amino acid sequence contains two potential N-linked glycosylation consensus sites, N404 and N424. Either enzymatic deglycosylation or tunicamycin reduced the cotransporter to its core molecular weight (113 kD). Glycosylation site single mutants expressed in oocytes ran as thick bands at 115 kD, consistent with the high-mannose glycoprotein. The double mutant produced the single thin 113-kD band seen in the deglycosylated cotransporter. Functional expression of cotransporters in Xenopus laevis oocytes revealed that the mutants displayed drastically decreased thiazide-sensitive (22)Na(+) uptake compared with wild-type NCC. Analysis of enhanced green fluorescence protein (EGFP)-tagged cotransporters demonstrated that this decrease in function is predominantly secondary to decreased surface expression. The elimination of glycosylation in the double mutant increased thiazide sensitivity by more than two orders of magnitude and also increased Cl(-) affinity. Thus, we have demonstrated that rNCC is N-glycosylated in vivo at two sites, that glycosylation is essential for efficient function and surface expression of the cotransporter, and that the elimination of glycosylation allows much greater access of thiazide diuretics to their binding site. PMID- 12538727 TI - Silencing of TonEBP/NFAT5 transcriptional activator by RNA interference. AB - TonEBP is a transcriptional activator that is expressed throughout development in many tissues and cell types. In the kidney medulla, TonEBP appears to be an important local regulator of differentiation by virtue of stimulating several genes. To study the function of TonEBP, two small interfering RNA (siRNA) duplexes were developed that reduced TonEBP expression effectively via RNA interference. The silencing lasted only 3 d after introduction of the TonEBP siRNA's. As expected, TonEBP-driven reporter gene expression and expression of the sodium/myo-inositol cotransproter (SMIT), aldose reductase (AR) and heat shock protein 70 (HSP70) mRNA were significantly decreased in cells where TonEBP expression was silenced. These data provide direct evidence that the SMIT, AR, and HSP70 genes are targets of TonEBP, although the potential role of other proteins, such as accessory proteins, cannot be excluded. The TonEBP-siRNA is an effective tool that should prove useful in the investigation of loss-of-function relationship in cells. PMID- 12538728 TI - Annexin II is present on renal epithelial cells and binds calcium oxalate monohydrate crystals. AB - Attachment of newly formed crystals to renal epithelial cells appears to be a critical step in the development of kidney stones. The current study was undertaken to identify potential calcium oxalate monohydrate (COM) crystal binding proteins on the surface of renal tubular cells. Apical membranes were prepared from confluent monolayers of renal epithelial cells (MDCKI line), and COM crystal affinity was used to isolate crystal-binding proteins that were then subjected to electrophoresis and electroblotting. Microsequencing of the most prominent COM crystal-binding protein (M(r) of 37 kD) identified it as annexin II (Ax-II). When exposed proteins on the surface of intact monolayers were biotinylated and then isolated using streptavidin agarose beads, Ax-II was detected, suggesting that at least a portion is exposed on the apical cell surface. Ax-II was not completely extracted by 0.1 M Na(2)CO(3), suggesting that at least a portion of cellular Ax-II is an intrinsic membrane-bound protein. Using confocal immunofluorescence microscopy, Ax-II was visualized together with Caveolin-1 (Cav-1) on the apical membrane of intact MDCKI cells. Cells pretreated with a monoclonal anti-Ax-II antibody bound significantly fewer COM crystals, whereas anti-LDL receptor antibody did not decrease COM binding, further suggesting a functional role for Ax-II during adhesion of crystals to intact cells. These results suggest that Ax-II avidly binds COM crystals and is present on the apical surface of MDCKI cells. Therefore, in the intact nephron, Ax-II could mediate adhesion of COM crystals to cells, and altered exposure of Ax-II on the surface of renal tubular cells could promote crystal retention and possibly kidney stone formation. PMID- 12538729 TI - Fabry disease in mice is associated with age-dependent susceptibility to vascular thrombosis. AB - Fabry disease is an X-linked lysosomal storage disorder due to deficiency of alpha-galactosidase A (GLA) activity that results in the widespread accumulation of neutral glycosphingolipids. Renal failure, neuropathy, premature myocardial infarction, and stroke occur in patients with this condition primarily due to deposition of glycosphingolipids in vascular endothelial cells. The clinical consequences of Fabry disease suggest that vascular thrombosis may play a prominent role in the pathogenesis of this disease; however, the vasculopathy associated with Fabry disease has not been extensively studied. To determine if mice genetically deficient in Gla are susceptible to vascular thrombosis, a photochemical carotid injury model was used to induce occlusive thrombosis. In this model, Gla-/0 mice displayed a progressive age-dependent shortening of the time to occlusive thrombosis after vascular injury that correlated with progressive accumulation of globotriasylceramide (Gb3) in the arterial wall. Bone marrow transplantation from Gla-/0 to Gla+/0 mice and from Gla+/0 to Gla-/0 mice did not change the thrombotic phenotype of the host. These studies reveal a potent vascular prothrombotic phenotype in Gla-deficient mice and suggest that antithrombotic therapies as well as therapies designed to reduce the vascular accumulation of Gb3 may have beneficial effects on thrombotic complications in patients with Fabry disease. PMID- 12538730 TI - Pressure natriuresis in AT(2) receptor-deficient mice with L-NAME hypertension. AB - AT(2) receptor-disrupted (AT(2) -/-) mice provide a unique opportunity to investigate the cardiovascular and BP-related effects of NO depletion. This study compared the pressure-diuresis-natriuresis relationship in (AT(2) -/-) and wild type (AT(2) +/+) mice after treating the animals with L-NAME (130 mg/kg body wt per day) for 1 wk. L-NAME increased mean arterial pressure (MAP) more in AT(2) -/ than in AT(2) +/+ mice (118 +/- 2 versus 108 +/- 4 mmHg). This difference occurred even though L-NAME-treated AT(2) +/+ mice had a greater sodium excretion than AT(2) -/- mice (10.9 +/- 0.5 versus 8.0 +/- 1.0 micro mol/h). The pressure natriuresis relationship in conscious AT(2) -/- mice was shifted rightward compared with controls. RBF was decreased in AT(2) -/- compared with AT(2) +/+ mice. L-NAME decreased RBF in these mice further from 4.08 +/- 0.43 to 2.79 +/- 0.15 ml/min per g of kidney wt. GFR was not significantly different between AT(2) +/+ and AT(2) -/- mice (1.09 +/- 0.08 versus 1.21 +/- 0.09 ml/min per g of kidney wt). L-NAME reduced GFR in AT(2) -/- to 0.87 +/- 0.07 ml/min per g of kidney wt. Fractional sodium (FE(Na)) and water (FE(H2O)) curves were shifted more strongly to the right by L-NAME in AT(2) -/- mice than in AT(2) +/+ mice. AT(1) receptor blocker treatment lowered BP in both L-NAME-treated strains to basal values. It is concluded that the AT(1) receptor plays a key role in the impaired renal sodium and water excretion induced by NO synthesis blockade. Changes in RBF, GFR, and tubular sodium and water reabsorption are involved and may be also responsible for the greater BP increase in L-NAME-treated AT(2) -/- mice. PMID- 12538731 TI - The apolipoprotein e knockout mouse: a model documenting accelerated atherogenesis in uremia. AB - Rodents do not develop spontaneous atherosclerosis. Currently, there is no good animal model to study the effect of uremia on atherosclerosis. This study evaluated whether apolipoprotein E knockout (Apoe-/-) mice are useful to study the effect of renal dysfunction on cardiovascular risk. Apoe-/- mice have decreased serum apolipoprotein E and exhibit lipid abnormalities and atherosclerosis even on a low-cholesterol diet. Ten-wk-old Apoe-/- mice were subtotally nephrectomised (SNX Apoe-/-; n = 8), uninephrectomised (UNX Apoe-/-; n = 5), or sham-operated (sham Apoe-/-; n = 5) and compared with their genetic controls (SNX C57/BL6; UNX C57/BL6; sham C57/BL6). After 12 wk, BP was measured intraarterially, blood samples were taken, and the experiment was terminated by perfusion fixation. The heart weight was determined, and quantitative morphologic analysis of intramyocardial arteries and aortic changes was performed. At the end of the experiment, heart weight and relative left ventricular weight were comparable in all groups. Intraarterial BP was somewhat higher in Apoe-/- mice compared with controls. Baseline serum cholesterol and triglyceride levels were higher in Apoe-/- mice than in C57/BL6. Atherosclerotic plaques were not present in sham or UNX C57/BL6, but minor plaque formation was noted in some SNX control animals. In contrast, beginning plaques were seen even in untouched Apoe-/- mice, and strikingly increased plaque formation was noted in UNX and SNX Apoe-/- mice. Maximal plaque diameter (cross-section) was 37 +/- 74 micro m in SNX C57/BL6, 191 +/- 90 micro m in sham Apoe-/-, 323 +/- 66 micro m in UNX Apoe-/-, and 457 +/- 17 micro m in SNX Apoe-/-. The plaque morphology corresponded with that of early plaques characterized by foam cells and virtual absence of lymphocytes or smooth muscle cell infiltration. In conclusion, even mild renal dysfunction, i.e., after uninephrectomy, causes a dramatic increase in plaque size and aggressive morphology (foam cell rich soft plaques) in the animal model of the Apoe-/- mouse. PMID- 12538732 TI - Bacterial CpG-DNA aggravates immune complex glomerulonephritis: role of TLR9 mediated expression of chemokines and chemokine receptors. AB - Immune complex glomerulonephritis (GN) often deteriorates during infection with viruses and bacteria that, in contrast to mammals, have DNA that contains many unmethylated CpG motifs. Balb/c mice with horse apoferritin-induced GN (HAF-GN) were treated with either saline, CpG-oligodeoxynucleotides (ODN), or control GpC ODN. Only CpG-ODN exacerbated HAF-GN with an increase of glomerular macrophages, which was associated with massive albuminuria and increased renal MCP-1/CCL2, RANTES/CCL5, CCR1, CCR2, and CCR5 mRNA expression. CpG-ODN induced a Th1 response as indicated by serum anti-HAF IgG(2a) titers, mesangial IgG(2a) deposits, and splenocyte IFN-gamma secretion. Messenger RNA for the CpG-DNA receptor Toll-like reeptor 9 (TLR9) was present in kidneys with HAF-GN but not in normal kidneys. The source of TLR9 mRNA in HAF-GN could be infiltrating macrophages or intrinsic renal cells, e.g., mesangial cells; but, in vitro, only murine J774 macrophages expressed TLR9. In J774 cells, CpG-ODN induced the chemokines MCP-1/CCL2 and RANTES/CCL5 and the chemokine receptors CCR1 and CCR5. It is concluded that CpG DNA can aggravate preexisting GN via a shift toward a Th1 response but also by a novel pathway involving TLR9-mediated chemokine and chemokine receptor expression by macrophages, which may contribute to the enhanced glomerular macrophage recruitment and activation. This mechanism may be relevant during infection triggered exacerbation of human immune-complex GN and other immune-mediated diseases in general. PMID- 12538733 TI - Enhanced expression of the CD71 mesangial IgA1 receptor in Berger disease and Henoch-Schonlein nephritis: association between CD71 expression and IgA deposits. AB - ABSTRACT. IgA nephropathy (IgA-N) that comprises Berger disease and Henoch Schonlein Purpura (HSP) nephritis is defined by mesangial IgA deposits. Recently, this group has characterized a new receptor for IgA, the transferrin receptor (CD71), expressed on mesangial cells. To assess whether CD71 was involved in the pathogenesis of IgA-N, its expression was analyzed together with IgA deposits on 16 kidney biopsies from 16 patients with Berger disease (n = 4) or HSP (n = 12). These biopsies were compared with 17 kidney biopsies of a group of 15 patients (control group) with other glomerulonephritis, including systemic lupus erythematosus, poststreptococcal acute glomerulonephritis, membranoproliferative glomerulonephritis, steroid-sensitive minimal change nephrotic syndrome, steroid resistant idiopathic nephrotic syndrome with focal and segmental glomerulosclerosis, and persistent and isolated proteinuria with minimal change on kidney biopsy. In this control group, IgA deposits could be observed in eight kidney biopsies of seven patients. These biopsies were also compared with normal kidney specimens (normal group). In normal kidney, it was found that CD71 was linearly expressed on tubular epithelium but was either not expressed or very dimly in glomeruli. In contrast, CD71 was strongly expressed in 105 of the 107 glomeruli of the kidney biopsies from the IgA-N group. For the control group, it was found that expression of CD71 in glomeruli was correlated to the presence of IgA deposits. Indeed, among the 87 glomeruli of nine kidney biopsies (eight patients) without IgA fixation, 78 exhibited no CD71 expression and nine exhibited a very dim one. On the other hand, all 49 glomeruli of the eight kidney biopsies (seven patients) in which IgA deposits were detected exhibited CD71 expression (P < 10(-4)). Performance of dual-labeling studies with confocal microscopy on kidney biopsies of IgA-N patients demonstrated that most of the IgA deposits co-localized with CD71. It was also demonstrated that the intensity of the expression of CD71 was not linked to the intensity of clinical or biologic findings but to the intensity of cellular proliferation in both IgA-N and control groups. These results show that mesangial CD71 expression is not specific to IgA N. However, the association between IgA deposits and CD71 expression and their co localization in the mesangium provide strong evidence that CD71 is a major IgA receptor on mesangial cells. PMID- 12538734 TI - Blockade of p38alpha MAPK ameliorates acute inflammatory renal injury in rat anti GBM glomerulonephritis. AB - The p38 mitogen-activated protein kinase (MAPK) pathway is a pro-inflammatory signal transduction pathway. The aim of this study was to examine the role of this pathway in acute renal inflammation. Immunostaining localized components of the p38 MAPK pathway (p38alpha, p-p38, p-ATF-2) in normal glomeruli, to podocytes, and occasional endothelial cells. This study identified an eightfold increase in glomerular activation of p38 MAPK (phosphorylated p38, p-p38) within 3 h of the induction of rat anti-glomerular basement membrane (GBM) glomerulonephritis and localized p-p38 and p-ATF-2 to infiltrating neutrophils, with increased staining of podocytes and endothelial cells. The relevance of these findings to human acute inflammatory renal disease was determined by examination of biopsy specimens. In patients with post-infectious glomerulonephritis, there was an increased number of positive p-p38 glomerular cells, including p-p38 staining of infiltrating neutrophils, compared with normal human kidney. In rats, administration of a specific p38 MAPK inhibitor, NPC 31145, before induction of anti-GBM disease prevented a loss of renal function and substantially reduced proteinuria. The reduction in renal injury was attributed to a 55% reduction in glomerular neutrophil infiltration and a 68% reduction in platelet accumulation. This was associated with an abrogation of glomerular P-selectin immunostaining and inhibition of glomerular P-selectin gene expression. In summary, this study has localized the components of the p38 MAPK pathway to cells in normal and diseased rat and human kidney and identified a number of important mechanisms by which signaling through the p38 MAPK pathway induces inflammatory renal disease. Blockade of the p38 pathway may be a novel therapeutic strategy for the treatment of acute renal inflammation. PMID- 12538735 TI - Alternatively used promoters and distinct elements direct tissue-specific expression of nephrin. AB - Nephrin, an essential component of the glomerular ultrafilter, the slit diaphragm, has also been found to be expressed in the central nervous system and pancreas. This study examined the regulation of the nephrin gene by analyzing the expression of different length nephrin promoter-lacZ reporter constructs in transgenic mice. An upstream segment between -4 kb and -4 bp was shown to be sufficient for driving expression in all three tissues. Surprisingly, a 5.7-kb construct lacking the transcription initiation site and the immediate upstream region of the gene could drive expression in the central nervous system. This led to the identification of a novel, alternatively used exon 1B located 1871 bp upstream of the ATG codon of the previously known first exon, now termed exon 1A. The existence and functionality of exon 1B was verified in nephrin knockout mice in which exon 1A is deleted. Deletion of exon 1B and its immediate surrounding sequence, introduced in the 4-kb promoter-lacZ reporter construct, abolished the expression of the transgene in pancreas and spinal cord but not in kidney and brain in transgenic mice. Analysis of five promoter-reporter gene constructs showed that regulatory elements driving expression encoded by exon 1A in kidney and brain are localized in the region between -4 kb and 2.1 kb. PMID- 12538736 TI - Molecular cytogenetic aberrations in autosomal dominant polycystic kidney disease tissue. AB - Autosomal dominant polycystic kidney disease (ADPKD) is a genetically heterogeneous disorder characterized by focal cyst formation from any part of the nephron. The molecular bases include germinal mutation of either PKD1 or PKD2 genes, enhanced expression of several protooncogenes, alteration of the TGF alpha/EGF/EGF receptor (EGFR) axis, and disturbed regulation of proliferative/apoptosis pathways. To identify new locations of ADPKD related oncogenes and/or tumor suppressor genes (TSG), comparative genomic hybridization (CGH) and loss of heterozygosity (LOH) analyses were performed for a series of individual cysts (n = 24) from eight polycystic kidneys. By CGH, imbalances were detected predominantly on chromosomes 1p, 9q, 16p, 19, and 22q in all tissues. DNA copy number gain was seen on chromosomes 3q and 4q in five samples. The CGH data were supplemented by LOH analysis using 83 polymorphic microsatellite markers distributed along chromosomes 1, 9, 16, 19, and 22. The highest frequency of LOH was found on the 1p35-36 and 16p13.3 segments in cysts from seven samples. Allelic losses on 9q were detected in six, whereas deletions at 19p13 and 22q11 bands were observed in three polycystic kidneys. These results indicate that the deleted chromosomal regions may contain genes important in ADPKD initiation and progression. PMID- 12538737 TI - ETA receptor blockade induces tubular cell proliferation and cyst growth in rats with polycystic kidney disease. AB - Tissue concentrations of ET-1 are markedly elevated in the kidneys of Han:Sprague Dawley (Han:SPRD) rats, a model of human autosomal dominant polycystic kidney disease (ADPKD). This study analyzed whether disease progression might be attenuated by endothelin receptor antagonists. Heterozygous Han:SPRD rats received an ETA receptor antagonist (LU 135252), a combined ETA/ETB receptor antagonist (LU 224332), or placebo for 4 mo. Glomerulosclerosis, protein excretion, and GFR remained unchanged, whereas interstitial fibrosis was enhanced by both compounds. BP was not reduced by both compounds in Han:SPRD rats. Renal blood flow (RBF) decreased in ADPKD rats treated with the ETA receptor antagonist. Long-term ETA receptor blockade furthermore increased markedly the number of renal cysts (ADPKD rats, 390 +/- 119 [cysts/kidney section +/- SD]; LU 135252-treated APKD rats, 1084 +/- 314; P < 0.001), cyst surface area (ADPKD rats, 7.97 +/- 2.04 [% of total section surface +/- SD]; LU 135252-treated ADPKD rats, 33.83 +/- 10.03; P < 0.001), and cell proliferation of tubular cells (ADPKD rats, 42.2 +/- 17.3 [BrdU-positive cells/1000 cells]; LU 135252-treated ADPKD rats, 339.4 +/- 286.9; P < 0.001). The additional blockade of the ETB receptor attenuated these effects in Han:SPRD rats. Both endothelin receptor antagonists had no effect on BP, protein excretion, GFR, and kidney morphology in Sprague Dawley rats without renal cysts. It is concluded that ETA receptor blockade enhances tubular cell proliferation, cyst number, and size and reduces RBF in Han:SPRD rats. This is of major clinical impact because endothelin receptor antagonists are upcoming clinically used drugs. PMID- 12538738 TI - Therapeutic role of TGF-beta-neutralizing antibody in mouse cyclosporin A nephropathy: morphologic improvement associated with functional preservation. AB - TGF-beta is believed to play a central role in the development of Cyclosporin A (CsA)-induced nephropathy. This study investigated the effects of 1D11, a murine pan-specific TGF-beta-neutralizing monoclonal antibody, in an ICR mouse model of chronic CsA nephropathy. Mice were administered a low-salt diet (0.01% sodium) for 1 wk followed by CsA treatment (30 mg/kg, subcutaneously, daily) for 4 wk. 1D11 was administered (2.5 mg/kg, intraperitoneally, 3 times/wk) beginning immediately after the termination of CsA dosing and continued through 8 wk. CsA caused extensive renal histopathologic alterations, including tubular damage, interstitial infiltrates and fibrosis, deposition of collagen III, and apoptosis of tubular epithelial cells. 1D11 ameliorated the CsA-induced histopathologic alterations, with significant reduction in collagen III expression and deposition. Additionally, elevated levels of mRNA encoding TGF-beta1 and TGF beta2 were significantly reduced. 1D11 also protected tubular epithelial cells from apoptosis by 48% (P < 0.05). In contrast, 13C4 (a control antibody) had no significant effect on any of the endpoints described above. Importantly, the effects of 1D11 on the CsA-induced morphologic alterations were followed by a reduction in serum creatinine level when compared with CsA mice treated with 13C4 (13C4, 0.45 +/- 0.09; 1D11, 0.30 +/- 0.08; P < 0.05) after 8 wk of treatment. Endothelial nitric oxide synthase (eNOS), inducible NOS (iNOS), nitrotyrosine, and tissue hypoxia were examined by immunostaining using specific antibodies. eNOS was significantly reduced in the endothelium of arterioles in the kidneys of mice treated with CsA, whereas iNOS was induced in the cortical tubules. Tissue hypoxia was found in both the arterioles and tubules, whereas nitrotyrosine was localized in the tubules. Administration of 1D11 improved tissue hypoxia and reduced nitrotyrosine formation. Moreover, the reciprocal changes in iNOS and eNOS expression were normalized by 1D11. This study demonstrates that 1D11 administration ameliorated morphologic alterations and preserved renal function in the context of existing chronic CsA nephropathy. PMID- 12538739 TI - Docosahexaenoic acid ameliorates murine ischemic acute renal failure and prevents increases in mRNA abundance for both TNF-alpha and inducible nitric oxide synthase. AB - This study demonstrates that intraperitoneal injections of DHA (all cis 4,7,10,13,16,19 docosahexaenoic acid C22: n-3) bound to bovine serum albumin ameliorate murine acute renal failure (ARF) induced by temporary occlusion of the renal artery. Three micromoles of DHA decreased serum creatinine (Scr) from 2.3 mg/dl to 1.1 mg/dl 24 h after reperfusion (n = 15; P < 0.05). Scr of the treated animals were significantly lower than controls throughout a 7-d time course. Although lower doses of DHA were less effective, higher doses were not more effective. Ribonuclease (RNase) protection assays showed that ischemia increased mRNA abundance for TNF-alpha and inducible nitric oxide synthase (iNOS) at 24 h. This increase was prevented by DHA administration. Because TNF-alpha and iNOS contribute to renal ischemic injury, their inhibition may contribute to DHA's salutary effect. In addition, the data may have therapeutic implications, because the DHA improves ARF even when administered at 4 h after reperfusion. PMID- 12538740 TI - Endothelin a receptor blockade and endothelin B receptor blockade improve hypokalemic nephropathy by different mechanisms. AB - Hypokalemia causes renal tubulointerstitial injury with an elevation in renal endothelin-1 (ET-1). It was hypothesized that hypokalemic tubulointerstitial injury is ameliorated by the blockade of ET-A receptors (ETA), whereas ET-B receptor (ETB) antagonism may exacerbate the injury, because ETB is thought to mediate vasodilation. Rats were fed a K(+)-deficient diet alone (LC) or with an ETA-selective antagonist ABT-627 (LA) or an ETB-selective antagonist A-192621 (LB) for 8 wk. Control rats were on a normal K(+) diet alone or with the ETA selective or ETB-selective antagonists. The severity of hypokalemia was not significantly different among LA, LB, and LC. LC developed tubulointerstitial injury with an elevation of renal preproET-1 mRNA level. There was an increase in tubular osteopontin expression, macrophage infiltration, collagen accumulation, and tubular cell hyperplasia. ETA blockade significantly ameliorated all parameters for renal injury in the cortex without suppressing local ET-1 and ETA expression. By contrast, ETB blockade significantly reduced local ET-1 and ETA expression and improved the injury to a similar extent in the cortex. In the medulla, ETA or ETB blockade only partially blocked renal injury. ETA blockade did not affect BP in normokalemic or hypokalemic rats. ETB blockade induced a BP elevation with a decrease in urinary Na(+) excretion in normokalemic but not in hypokalemic rats. These results indicate that ET-1 can mediate hypokalemic renal injury in two different ways: by directly stimulating ETA and by locally promoting endogenous ET-1 production via ETB. Thus, ETA as well as ETB blockade may be renoprotective in hypokalemic nephropathy. PMID- 12538741 TI - PTH pulsatility but not calcium sensitivity is restored after total parathyroidectomy with heterotopic autotransplantation. AB - In healthy humans, parathyroid hormone (PTH) is secreted via basal mode with superimposed oscillatory bursts every 8 to 12 min. Amplitude and frequency changes mediate the instantaneous response of the parathyroids to changes in ambient Ca(2+) concentrations. The parathyroid gland tetrad may be synchronized by autonomic innervation. This study investigated the effect of total parathyroidectomy and heterotopic autotransplantation of parathyroid tissue (PTX) on PTH secretion patterns in nine patients with end-stage renal disease. Intact PTH versus time concentration profiles were obtained early (1 to 8 wk, n = 4) or late (15 to 33 mo, n = 5) after PTX. In four patients late after PTX, Ca(2+) responsiveness of PTH secretion was additionally investigated by citrate and calcium clamp studies. The nonrandomness of plasma PTH fluctuations was assessed by the approximate entropy (ApEn) statistic, and secretion characteristics by multiparametric deconvolution analysis. Results were compared with those of matched normal subjects and chronic renal failure (CRF) patients without PTX. PTH burst frequency was 2.9 +/- 0.1 h(-1) early and 7 +/- 0.4 h(-1) late after PTX as compared with 8.1 +/- 0.4 h(-1) in CRF and 7 +/- 0.3 h(-1) in healthy controls. Fractional pulsatile PTH secretion was diminished after PTX (18 +/- 2%) compared with healthy controls (32 +/- 5%, P < 0.05) and CRF patients (25 +/- 4%, P = 0.05). The orderliness of PTH release was significantly reduced after PTX (ApEn: 1.59 +/- 0.03 versus 1.41 +/- 0.09 in healthy and 1.46 +/- 0.03 in CRF controls, P < 0.01). Acute hypocalcemia elicited a lesser increase in pulsatile PTH secretion in PTX patients (147 +/- 134%) than in the CRF (500 +/- 92%, P = 0.05) and healthy controls (1410 +/- 290%, P < 0.05), mainly due to a diminished mass of PTH secreted per burst. Pulsatile PTH secretion was also resistant to hypercalcemia, wherein the suppression of burst mass was significantly reduced compared with that in healthy controls. In conclusion, pulsatile PTH secretion is partially restored within 2 yr of PTX. However, the capacity of the autotransplanted tissue to adapt to changes in ionized calcium remains profoundly disturbed. PMID- 12538742 TI - Mortality differences by dialysis modality among incident ESRD patients with and without coronary artery disease. AB - It is unclear whether peritoneal dialysis (PD) compared with hemodialysis (HD) confers a survival advantage in end-stage renal disease (ESRD) patients with coronary artery disease (CAD). This hypothesis was tested in a national cohort of 107,922 patients starting dialysis therapy between May 1, 1995, and July 31, 1997. Data on patient characteristics were obtained from the Center for Medicare and Medicaid Services Medical Evidence Form (CMS) and linked to mortality data from the United States Renal Data System (USRDS). Patients were classified on the basis of CAD presence and followed until death or the end of 2 yr. Nonproportional Cox regression models estimated the relative risk (RR) of death for patients with and without CAD by dialysis modality using primarily the intent to-treat but also the as-treated approach. Diabetic patients (DM) and nondiabetic patients (non-DM) were analyzed separately. Among DM, patients with CAD treated with PD had a 23% higher RR (95% CI, 1.12 to 1.34) compared with similar HD patients, whereas patients without CAD receiving PD had a 17% higher RR (CI, 1.08 to 1.26) compared with HD. Among non-DM, patients with CAD treated with PD had a 20% higher RR (CI. 1.10 to 1.32) compared with HD patients, whereas patients without CAD had similar survival on PD or HD (RR = 0.99; CI, 0.93 to 1.05). The mortality risk for new ESRD patients with CAD differed by treatment modality. In both DM and non-DM, patients with CAD treated with PD had significantly poorer survival compared with HD. Whether differences in solute clearance and/or cardiac risk profiles between PD and HD may explain these findings deserves further investigation. PMID- 12538743 TI - Enalapril and losartan reduce sympathetic hyperactivity in patients with chronic renal failure. AB - The aim of this study was to compare the effects on BP and sympathetic activity of chronic treatment with an angiotensin (Ang)-converting enzyme (ACE) inhibitor and an AngII receptor blocker in hypertensive patients with chronic renal failure (CRF). In ten stable hypertensive CRF patients (creatinine clearance, 46 +/- 17 ml/min per 1.73 m(2)), muscle sympathetic nerve activity (MSNA), plasma renin activity (PRA), baroreceptor sensitivity, and 24-h ambulatory BP were measured in the absence of antihypertensive drugs (except diuretics) after 6 wk of enalapril (10 mg orally) and after 6 wk of losartan (100 mg orally). The order of the three phases was randomized. Normovolemia was controlled with diuretics and confirmed with extracellular fluid volume measurements throughout the study. Both enalapril and losartan reduced MSNA (from 33 +/- 10 to 27 +/- 13 and 27 +/- 13 bursts/min, respectively; P < 0.05) and average 24-h BP (from 141 +/- 8/93 +/- 8 to 124 +/- 9/79 +/- 8 and 127 +/- 8/81 +/- 9 mmHg; P < 0.01). PRA was not different during the treatments. The change in BP and the change in MSNA during the treatments were correlated (r = 0.70 and r = 0.63, respectively; both P < 0.05). Baroreceptor sensitivity was not affected by the treatments. This is the first study to compare the effects of ACE inhibition and AngII blockade on MSNA. In hypertensive CRF patients, enalapril and losartan equally reduced BP and MSNA. Differences in modes of action of the two drugs did not result in differences in effects on MSNA, supporting the view that AngII-mediated mechanisms contribute importantly in the pathogenesis of sympathetic hyperactivity in these patients. PMID- 12538744 TI - Prognostic value of myocardial perfusion studies in patients with end-stage renal disease assessed for kidney or kidney-pancreas transplantation: a meta-analysis. AB - The prognostic utility of myocardial perfusion studies (MPS) such as thallium scintigraphy and dobutamine stress echocardiography (DSE) for stratifying cardiac risk among candidates for kidney or kidney-pancreas transplantation is uncertain. This study is a meta-analysis to determine the prognostic significance of MPS results on future myocardial infarction (MI) and cardiac death (CD) in patients with end-stage renal disease (ESRD) assessed for kidney or kidney-pancreas transplantation. MEDLINE was searched using combinations of MeSH headings and text words for transplantation, coronary artery disease, prognosis, end-stage renal disease, and noninvasive cardiac testing (nuclear scintigraphy and DSE) for primary studies. Studies were included if they reported MPS results and cardiac events in patients assessed for kidney or kidney-pancreas transplantation. Methodologic study quality and outcome data were independently abstracted in duplicate by two researchers. The relative risks (RR) of MI and CD were calculated using a random effects model. Twelve articles met all inclusion criteria; 12 studies reported CD, and 9 reported MI. In eight studies, thallium scintigraphy was used (four with pharmacologic stress, four with exercise stress), whereas four used DSE. When compared with negative tests, positive tests had a significantly increased RR of MI (2.73 [95% CI, 1.25 to 5.97]; P = 0.01) and CD (2.92 [95% CI, 1.66 to 5.12]; P < 0.001). Subgroup analyses of studies of diabetic patients indicated that positive tests were associated with a RR of CD 3.95 (95% CI, 1.48 to 10.5; P = 0.006) and a RR of MI 2.68 (95% CI, 0.95 to 7.57; P = 0.06) when compared with negative tests. In studies evaluating mixed populations of diabetic and nondiabetic patients, positive tests were associated with a RR of CD 2.52 (95% CI, 1.25 to 5.08; P = 0.01) and with a RR of MI 2.79 (95% CI, 0.85 to 9.21; P = 0.09) when compared with a negative test. The presence of reversible defects was associated with an increased risk of MI in diabetic patients and of CD in both subgroups; fixed defects were associated with an increased risk of CD but not MI. It is concluded that positive MPS are useful in identifying patients with significantly increased risk of future MI and CD in both diabetic and nondiabetic ESRD patients. PMID- 12538745 TI - 15-Deoxyspergualin in patients with refractory ANCA-associated systemic vasculitis: a six-month open-label trial to evaluate safety and efficacy. AB - The combination of cyclophosphamide (CYC) and oral corticosteroids is effective in the majority of patients with antineutrophil cytoplasmic antibody-associated vasculitis (AASV), but it carries substantial risk of drug-related morbidity and mortality. New regimens are desired, especially in refractory cases. The immunosuppressant 15-deoxyspergualin (DSG) is effective in experimental autoimmune disease and transplantation as well as in acute kidney transplant rejection in humans. To assess the efficacy and safety of DSG, an open label multicenter trial was conducted in patients with AASV who were either unresponsive or had contraindications for standard immunosuppressants. Included were 19 cases of Wegener granulomatosis and one case of microscopic polyangiitis. Nine of them had received CYC shortly before study entry without apparent therapeutic success. DSG (0.5 mg/kg per d) was given for 2 to 3 wk until the WBC count dropped to 3000/ micro l followed by a rest until at least a WBC of 4000/ micro l was reached again. This was repeated up to six cycles. During the study, no other immunosuppressants besides steroids were allowed. Disease improvement during treatment with DSG was achieved in 70% of cases (six cases of complete remission; eight cases of partial remission). Leucopenia occurred in each patient in a regular pattern during the cycles and was transient without exception. No mortality or septicemia was observed. Mild to moderate infections mainly in the respiratory tract were observed but resolved under adequate treatment without sequel. It is concluded that treatment with DSG is successful in patients with refractory Wegener granulomatosis under careful monitoring of WBC count. PMID- 12538746 TI - Serial estimates of serum permeability activity and clinical correlates in patients with native kidney focal segmental glomerulosclerosis. AB - A serum or plasma factor in certain patients with focal segmental glomerulosclerosis (FSGS) has been found to increase glomerular albumin permeability (P(alb)) and causes proteinuria in experimental animals. High P(alb) is associated with recurrence of FSGS after transplantation, but serial studies of P(alb) activity in patients with native kidney FSGS have not been performed, and the relationship between P(alb) and remission of proteinuria is not known. This study was designed to determine P(alb) activity before, during, and after 24 wk of treatment with cyclosporine or placebo given as part of a randomized controlled trial in steroid-resistant FSGS patients with nephrotic range proteinuria. Pretreatment P(alb) averaged 0.36 +/- 0.22 and was not significantly different between treatment groups and was not altered during or after the test medication. There was no association between P(alb) activity and remission or relapse in proteinuria. The average P(alb) activity in native kidney FSGS was lower than in previously reported patients with posttransplant recurrence of the disease, and its level did not vary during the course of the study. The antiproteinuric effect of cyclosporine appeared independent of changes in P(alb). This finding is consistent with a direct effect of cyclosporine on glomerular barrier function and/or that within this group of patients the variations in proteinuria are not reflected in changes in Palb because of its limits in terms of reproducibility and responsiveness. PMID- 12538747 TI - Bioartificial kidney ameliorates gram-negative bacteria-induced septic shock in uremic animals. AB - The bioartificial kidney (BAK) consists of a conventional hemofiltration cartridge in series with a renal tubule assist device (RAD) containing 10(9) porcine renal proximal tubule cells. BAK replaces filtration, transport, and metabolic and endocrinologic activities of a kidney. Previous work in an acutely uremic dog model demonstrated that BAK ameliorated endotoxin (lipopolysaccharide [LPS])-induced hypotension and altered plasma cytokine levels. To further assess the role of BAK in sepsis in acute renal failure, dogs were nephrectomized and 48 h later administered intraperitoneally with 30 x 10(10) bacteria/kg of E. coli. One hour after bacterial administration, animals were placed in a continuous venovenous hemofiltration circuit with either a sham RAD without cells (n = 6) or a RAD with cells (n = 6). BP, cardiac output, heart rate, pulmonary capillary wedge pressure, and systemic vascular resistance were measured throughout the study. All animals tested were in renal failure, with blood urea nitrogen and serum creatinine concentrations greater than 60 and 6 mg/dl, respectively. RAD treatment maintained significantly better cardiovascular performance, as determined by arterial BP (P < 0.05) and cardiac output (P < 0.02), for longer periods than sham RAD therapy. Consistently, all sham RAD-treated animals, except one, expired within 2 to 9 h after bacterial administration, whereas all RAD treated animals survived more than 10 h. Plasma levels of TNF-alpha, IL-10, and C reactive protein (CRP) were measured during cell RAD and sham RAD treatment. IL 10 levels were significantly higher (P < 0.01) during the entire treatment interval in the RAD animals compared with sham controls. These data demonstrated in a pilot large animal experiment that the BAK with RAD altered plasma cytokine levels in acutely uremic animals with septic shock. This change was associated with improved cardiovascular performance and increased survival time. These results demonstrate that the addition of cell therapy to hemofiltration in an acutely uremic animal model with septic shock ameliorates cardiovascular dysfunction, alters systemic cytokine balance, and improves survival time. PMID- 12538748 TI - Electrocardiographic left ventricular hypertrophy in renal transplant recipients: prognostic value and impact of blood pressure and anemia. AB - Left ventricular hypertrophy (LVH) is an independent risk factor for death and cardiovascular disease in the general population and dialysis patients. However, the causes and consequences of LVH have not been well described in renal transplant recipients (RTR). A retrolective cohort study was conducted in 473 RTR who were alive and free of cardiac disease at 1 yr. LVH was defined using the Cornell electrocardiographic (EKG) criteria. A total of 416 patients had an interpretable first-year EKG (88%), and 284 had an interpretable fifth-year EKG (78% of 5-yr survivors). Baseline characteristics were similar in patients with and without EKG. Of 416 patients, 57 had LVH in the first year, whereas 38 of 284 patients had LVH in the fifth year, of which 18 cases were de novo. Baseline LVH was a risk factor for death (RR 1.9 [1.22, 3.22]) and congestive heart failure (CHF) (RR 2.27 [1.08, 4.81]) and was independent of other major prognostic variables. Persistent or de novo LVH in the fifth year predicted subsequent death (RR 2.15 [1.14,4.01]) and CHF (2.71 [1.17, 6.30]). Anemia and diastolic BP were independent risk factors for increasing Cornell voltage (a marker of LV mass) between first and fifth years. Systolic BP was the only predictor of de novo LVH at 5 yr. It seems that EKG LVH is a significant risk factor for death and CHF in RTR and that anemia and hypertension are risk factors for LV growth. Whether aggressive treatment of hypertension and anemia can improve outcomes merits further study. PMID- 12538749 TI - Insulin resistance and risk of chronic kidney disease in nondiabetic US adults. AB - This study examined the relationship of fasting serum glucose, insulin, C peptide, glycosylated hemoglobin A (HbA1c), and Homeostasis Model Assessment (HOMA)-insulin resistance to risk of chronic kidney disease (CKD) among 6453 persons without diabetes (fasting glucose <126 mg/dl and not taking diabetes medication) who participated in the Third National Health and Nutrition Examination Survey and were aged 20 yr or older. CKD was defined as an estimated GFR <60 ml/min per 1.73 m(2). The prevalence of CKD was significantly and progressively higher with increasing levels of serum insulin, C-peptide, HbA1c, and HOMA-insulin resistance. After adjustment for potential confounding variables, the odds ratio of CKD for the highest compared with the lowest quartile was 4.03 (95% confidence interval [CI], 1.81 to 8.95; P = 0.001), 11.4 (95% CI, 4.07 to 32.1; P < 0.001), 2.67 (95% CI, 1.31 to 5.46; P = 0.002), and 2.65 (95% CI, 1.25 to 5.62; P = 0.008) for serum insulin, C-peptide, HbA1c levels, and HOMA-insulin resistance, respectively. For a one SD higher level of serum insulin (7.14 micro U/ml), C-peptide (0.45 Deltamol/ml), HbA1c (0.52%), and HOMA-insulin resistance (1.93), the odds ratio (95% CI) of CKD was 1.35 (1.16 to 1.57), 2.78 (2.25 to 3.42), 1.69 (1.28 to 2.23), and 1.30 (1.13 to 1.50), respectively. These findings combined with knowledge from previous studies suggest that the insulin resistance and concomitant hyperinsulinemia are presented in CKD patients without clinical diabetes. Further studies into the causality between insulin resistance and CKD are warranted. PMID- 12538750 TI - Adjustment for comorbidity in studies on health status in ESRD patients: which comorbidity index to use? AB - Health status can be an important outcome in studies on patients with end-stage renal disease (ESRD). In these studies, adjustment for prognostic factors, such as comorbidity, often has to be made. None of the comorbidity indices that are commonly used in research on ESRD patients has been validated for studies on health status. This study evaluated three existing indices (Khan, Davies, and Charlson) and four indices specifically developed for use in studies on health status. In a large prospective multi-center study (NECOSAD-2), new ESRD patients were included (n = 1041). Comorbidity was assessed at the start of dialysis. Health status was assessed with the physical and mental component summary score of the SF-36 (PCS and MCS), the symptoms dimension of the KDQOL-SF, and the Karnofsky Scale. Patient data were randomly allocated to a modeling or a testing set. The new indices were developed in the modeling set. The three existing indices explained about the same percentage of variance in the PCS (7 to 8%), MCS (1 to 3%), symptoms (2 to 4%), and Karnofsky (10 to 12%). The new indices performed better than the existing indices in the modeling population (13% PCS, 10% MCS, 10% symptoms, 18% Karnofsky), but not in the testing population (8% PCS, 1% MCS, 3% symptoms, 8% Karnofsky). Individual comorbidities explained more variance in PCS (10 to 15%), MCS (1 to 7%), symptoms (6 to 11%), and Karnofsky (11 to 18%) than comorbidity indices. The Khan, Davies, and the Charlson indices will adjust to the same extent for the potential confounding effect of comorbidity in studies with health status as an outcome. Separate comorbidity diagnoses will adjust best for comorbidity. PMID- 12538751 TI - A propensity analysis of late versus early nephrologist referral and mortality on dialysis. AB - Previous studies have analyzed the association between late versus early nephrologist referral (LR, ER) and poor clinical outcomes in patients with end stage renal disease. We sought to determine whether these poor outcomes were causally related to LR, or whether LR was a proxy for poorer access to health care in general. An inception cohort of incident dialysis patients enrolled in the New Jersey Medicare or Medicaid programs was identified. Using a large number of demographic, clinical, and health care utilization covariates, propensity scores (PS) were then calculated to predict whether a given patient had been seen by a nephrologist at 90 d before first dialysis. Cox proportional hazards models were then built to test the association between timing of nephrologist referral and mortality during the first year of dialysis, using PS adjustment and matching to determine whether this association was confounded by other measures of reduced healthcare utilization. Neither adjustment for PS (HR = 1.31; 95% CI, 1.17 to 1.47) nor matching (HR = 1.40; 95% CI, 1.23 to 1.59) materially changed the initial 36% excess mortality in LR compared with ER patients (HR = 1.36; 95% CI, 1.22 to 1.51). Excess mortality among LR was limited to the first 3 mo of dialysis (HR = 1.75; 95% CI, 1.48 to 2.08) but not present thereafter (HR = 1.03; 95% CI, 0.84 to 1.25). Late nephrologist referral is an independent risk factor for early death on dialysis, even after controlling for other indicators of healthcare utilization. Further research is needed to identify patients at particular risk so that interventions to prevent early deaths on dialysis in LR patients can be developed and tested. PMID- 12538752 TI - Donor tissue characteristics influence cadaver kidney transplant function and graft survival but not rejection. AB - Acute injury and age are characteristics of transplanted tissue that influence many aspects of the course of a renal allograft. The influence of donor tissue characteristics on outcomes can be analyzed by studying pairing, the extent to which two kidneys retrieved from the same cadaver donor manifest similar outcomes. Pairing studies help to define the relative role of donor-related factors (among pairs) versus non-donor factors (within pairs). This study analyzed graft survival for 220 pairs of cadaveric kidneys for the similarity of parameters reflecting function and rejection. It also examined whether the performance of one kidney was predicted by the course of its "mate," the other kidney from that donor. Parameters reflecting function showed sustained pairing posttransplantation, as did graft survival. In contrast, measures of rejection strongly affected survival but showed no pairing. Surprisingly, the survival of a kidney was predicted by the early performance of its mate, an observation we term the "mate effect." Six-month graft survival and renal function were reduced in grafts for which the mate kidney displayed any criteria for functional impairment (dialysis dependency, low urine output [/= 400 micro mol/L), even for kidneys which themselves lacked those criteria. Rejection measures did not demonstrate the mate effect. In conclusion, kidney transplant function is strongly linked to donor-related factors (age, brain death). In contrast, rejection affects survival and function, but it is not primarily determined by the characteristics of the donor tissue. Graft survival reflects both of these influences. PMID- 12538753 TI - Dialysis, kidney transplantation, or pancreas transplantation for patients with diabetes mellitus and renal failure: a decision analysis of treatment options. AB - Patients with type 1 diabetes mellitus and end-stage renal disease may remain on dialysis or undergo cadaveric kidney transplantation, living kidney transplantation, sequential pancreas after living kidney transplantation, or simultaneous pancreas-kidney transplantation. It is unclear which of these options is most effective. The objective of this study was to determine the optimal treatment strategy for type 1 diabetic patients with renal failure using a decision analytic Markov model. Input data were obtained from the published medical literature, the United Network for Organ Sharing registry, and patient interviews. The outcome measures were life expectancy (in life-years [LY]) and quality-adjusted life expectancy (in quality-adjusted life-years [QALY]). Living kidney transplantation was associated with 18.30 LY and 10.29 QALY; pancreas after kidney transplantation, 17.21 LY and 10.00 QALY; simultaneous pancreas kidney transplantation, 15.74 LY and 9.09 QALY; cadaveric kidney transplantation, 11.44 LY and 6.53 QALY; dialysis, 7.82 LY and 4.52 QALY. The results were sensitive to the value of several key variables. Simultaneous pancreas-kidney transplantation had the greatest life expectancy and quality-adjusted life expectancy when living kidney transplantation was excluded from the analysis. These data indicate that living kidney transplantation is associated with the greatest life expectancy and quality-adjusted life expectancy for type 1 diabetic patients with renal failure. Treatment strategies involving pancreas transplantation should be considered for patients with frequent metabolic complications of diabetes and for those patients who favor kidney-pancreas transplantation over kidney transplantation alone. For patients without a living donor, simultaneous pancreas-kidney transplantation is associated with the greatest life expectancy. PMID- 12538754 TI - Eye and kidney: from clinical findings to genetic explanations. PMID- 12538755 TI - Glomerular hyperfiltration and the salt paradox in early [corrected] type 1 diabetes mellitus: a tubulo-centric view. AB - Diabetes mellitus contributes greatly to morbidity, mortality, and overall health care costs. In major part, these outcomes derive from the high incidence of progressive kidney dysfunction in patients with diabetes making diabetic nephropathy a leading cause of end-stage renal disease. A better understanding of the early dysfunctions observed in the diabetic kidney may permit the development of new strategies to prevent diabetic nephropathy. This review proposes a "tubulo centric" view of glomerular function in early type I diabetes mellitus. The following are particularly discussed (1) the primary role of an increase in reabsorption by the proximal tubule in early glomerular hyperfiltration, (2) the role of sodium-glucose cotransport and tubular growth under these conditions, and (3) the primary role of reabsorption by the proximal tubule for the paradoxical relationship between dietary salt and glomerular filtration rate. Finally, an outline is presented of potential therapeutic implications for the prevention of diabetic kidney disease. PMID- 12538756 TI - The thiazide-sensitive na-cl cotransporter and human disease: reemergence of an old player. PMID- 12538757 TI - Circulating permeability factors in the nephrotic syndrome: a fresh look at an old problem. PMID- 12538758 TI - The nuclear receptor superfamily. PMID- 12538759 TI - Nuclear RNA export. AB - Eukaryotic cells export several different classes of RNA molecule from the nucleus, where they are transcribed, to the cytoplasm, where the majority participate in different aspects of protein synthesis. It is now clear that these different classes of RNA, including rRNAs, tRNAs, mRNAs and snRNAs, are specifically directed into distinct but in some cases partially overlapping nuclear export pathways. All non-coding RNAs are now known to depend on members of the karyopherin family of Ran-dependent nucleocytoplasmic transport factors for their nuclear export. In contrast, mRNA export is generally mediated by a distinct, Ran-independent nuclear export pathway that is both complex and, as yet, incompletely understood. However, for all classes of RNA molecules, nuclear export is dependent on the assembly of the RNA into the appropriate ribonucleoprotein complex, and nuclear export therefore also appears to function as an important proofreading mechanism. PMID- 12538760 TI - Plastic adult stem cells: will they graduate from the school of hard knocks? AB - Notwithstanding the fact that adult bone marrow cell engraftment to epithelial organs seems a somewhat uncommon event, there is no doubt it does occur, and under appropriate conditions of a strong and positive selection pressure these cells will expand clonally and make a significant contribution to tissue replacement. Likewise, bone-marrow-derived cells can be amplified in vitro and differentiated into a multitude of tissues. These in essence are the goals of regenerative medicine using any source of stem cells, be it embryonic or adult. Despite such irrefutable evidence of what is possible, a veritable chorus of detractors of adult stem cell plasticity has emerged, some doubting its very existence, motivated perhaps by more than a little self-interest. The issues that have led to this state of affairs have included the inability to reproduce certain widely quoted data, one case where the apparent transdifferentiation was due to contamination of the donor tissue with haematopoietic cells and, most notoriously, extrapolating from the behaviour of embryonic stem cells to suggest that adult bone marrow cells simply fuse with other cells and adopt their phenotype. While these issues need resolving, slamming this whole new field because not everything is crystal clear is not good science. The fact that a phenomenon is quite rare in no way mitigates against its very existence: asteroid collisions with the Earth are rare, but try telling the dinosaurs they do not occur! When such events do occur (transdifferentiation or collision), they certainly can make an impact. PMID- 12538761 TI - hRUL138, a novel human RNA-binding RING-H2 ubiquitin-protein ligase. AB - Cellular as well as viral RNAs are usually found complexed with proteins. In an attempt to identify proteins that interact with transcripts of hepatitis B virus (HBV), a DNA virus that replicates through reverse transcription, a partial cDNA was isolated from a human cDNA expression library whose gene product bound to an HBV-derived RNA. Using an overlapping clone from a molecular hybridization screen a full-length cDNA was assembled. It contained a large open reading frame for a 1208 amino-acid protein of 138 kDa identical to the hypothetical product of the KIAA0675 clone. Closely related sequences are present in mouse cDNA libraries but not in the genomes of lower organisms. The protein sequence contained no known RNA-binding domain and, apart from a probable coiled-coil domain, the only significant homology involved a complete RING-H2 motif. This suggested that the protein might be a novel RNA-binding RING-dependent ubiquitin-protein ligase or E3 enzyme. A motif critical for RNA binding was experimentally mapped to a central Lys-rich region. Binding specificity is either broad or the protein has as yet unknown physiological targets; hence, at present, a potential importance for HBV biology remains open. The RING-H2 domain was functional in and essential for self- and trans-ubiquitylation in vitro and for proteasome-mediated turnover of the protein in vivo. We therefore termed it hRUL138 for human RNA-binding ubiquitin ligase of 138 kDa. hRUL138 mRNAs are expressed at low levels in most tissues. GFP-tagged hRUL138 derivatives were found associated with cytoplasmic structures, possibly the ER, but excluded from the nucleus. The combined presence of RNA binding and E3 activity in hRUL138 raises the possibility that both are mechanistically linked. PMID- 12538762 TI - Analysis of Bub3 spindle checkpoint function in Xenopus egg extracts. AB - The spindle checkpoint delays the onset of anaphase if there are any defects in the interactions between spindle microtubules and kinetochores. This checkpoint has been reconstituted in vitro in Xenopus egg extracts, and here we use antibodies to Xenopus Bub3 (XBub3) to show that this protein is required for both the activation and the maintenance of a spindle checkpoint arrest in egg extracts. We detect two forms of XBub3 in egg extracts and find both to be complexed with the XBub1 and XBubR1 kinases. Only one form of XBub3 is apparent in Xenopus tissue culture (XTC) cells, and localisation studies reveal that, unlike the Mad proteins, which are concentrated at the nuclear periphery, XBub3 is diffusely localised throughout the nucleus during interphase. During early prophase it is recruited to kinetochores, where it remains until chromosomes align at the metaphase plate. We discuss the mechanism by which our alpha-XBub3 antibodies interfere with the checkpoint and possible roles for XBub3 in the spindle checkpoint pathway. PMID- 12538763 TI - ABA- and cADPR-mediated effects on respiration and filtration downstream of the temperature-signaling cascade in sponges. AB - Recently, the thermosensing pathway in sponges (Porifera) was elucidated. The thermosensor triggering this cascade is a heat-activated cation channel, with the phytohormone abscisic acid (ABA), cyclic ADP-ribose (cADPR) and calcium acting as intracellular messengers, similarly to the drought-stress signaling cascade in higher plants. Here, we investigated the functional effects downstream of the temperature-signaling pathway in Axinella polypoides (Porifera, Demonspongiae). Short-term stimulation followed by long-term depression of amino acid incorporation, oxygen consumption and water filtration were observed after exposure of the sponge to a brief heat stress or to micromolar ABA. These effects could be prevented by the targeted interruption of the signaling pathway either at the level of the cation channel thermosensor or at the level of the cADPR induced intracellular calcium increase. Moreover, release of cyclase activity into the sea water and generation of extracellular cADPR were observed following brief heat stress. Intact sponge cells were sensitive to extracellular cADPR and addition of purified cyclase increased sponge respiration similarly to heat stress. This is the first observation of functional effects exerted on Metazoa by the phytohormone ABA: conservation of the ABA/cADPR stress-signaling cascade points to its early evolution in a common precursor of modern Metazoa and Metaphyta. The functional effects induced by extracellular cyclase/cADPR suggest an evolutionary origin of cADPR as an ancient stress hormone in Porifera. PMID- 12538765 TI - Mitochondrial and nucleocytoplasmic targeting of O-linked GlcNAc transferase. AB - O-linked GlcNAc transferase (OGT) mediates a novel glycan-dependent signaling pathway, but the intracellular targeting of OGT is poorly understood. We examined the localization of OGT by immunofluorescence microscopy, subcellular fractionation and immunoblotting using highly specific affinity-purified antisera. In addition to the expected nuclear localization, we found that OGT was highly concentrated in mitochondria. Since the mitochondrial OGT (103 kDa) was smaller than OGT found in other compartments (116 kDa) we reasoned that it was one of two predicted splice variants of OGT. The N-termini of these isoforms are unique; the shorter form contains a potential mitochondrial targeting sequence. We found that when epitope-tagged, the shorter form (mOGT; 103 kDa) concentrated in HeLa cell mitochondria, whereas the longer form (ncOGT; 116 kDa) localized to the nucleus and cytoplasm. The N-terminus of mOGT was essential for proper targeting. Although mOGT appears to be an active transferase, O-linked GlcNAc modified substrates do not accumulate in mitochondria. Using immunoelectron microscopy and mitochondrial fractionation, we found that mOGT was tightly associated with the mitochondrial inner membrane. The differential localization of mitochondrial and nucleocytoplasmic isoforms of OGT suggests that they perform unique intracellular functions. PMID- 12538764 TI - A role for glycogen synthase kinase-3 in mitotic spindle dynamics and chromosome alignment. AB - Glycogen synthase kinase-3 (GSK-3) is a conserved, multifunctional kinase that is constitutively active in resting cells, and inactivated through phosphorylation by protein kinase B (PKB). We have investigated the temporal and spatial control of GSK-3 phosphorylation during the cell cycle in mammalian cells. We show that GSK-3 is present along the length of spindle microtubules and that a fraction of GSK-3 is phosphorylated during mitosis. Phospho-GSK-3 is abundant at the centrosomes and spindle poles but absent from other areas of the spindle. GSK-3 phosphorylation occurs concomitantly with the appearance of phosphorylated and active PKB at the centrosome, which suggests that PKB is the kinase responsible for phosphorylating and inactivating GSK-3 at the centrosome during mitosis. We demonstrate that lithium and two structurally distinct inhibitors of GSK-3 promote defects in microtubule length and chromosomal alignment during prometaphase. Treated cells contain mono-oriented chromosomes concentrated at the plus ends of astral microtubules, which are longer than in untreated cells. Live microscopy of cells expressing Histone-2B-GFP confirms that the inhibition of GSK 3 suppresses mitotic chromosome movement and leads to a prometaphase-like arrest. We propose that GSK-3 is regulated in a temporal and spatial manner during mitosis and, through controlling microtubule dynamics, plays an important role in chromosomal alignment on the metaphase plate. PMID- 12538766 TI - Multiple cytoplasmic signals direct the intracellular trafficking of chicken kidney AE1 anion exchangers in MDCK cells. AB - AE1/F(c) receptor chimeras have been used to define the sequences that direct the basolateral sorting, recycling and cytoskeletal association of the chicken AE1-4 anion exchanger in MDCK cells. These analyses revealed that amino acids 1-63 of AE1-4 were sufficient to redirect a cytoplasmic tailless murine IgG F(c)RII B2 receptor from the apical to the basolateral membrane of MDCK cells, where F(c)1 63 associated with elements of the actin cytoskeleton. In contrast to F(c)1-63, chimeras containing amino acids 1-37 (F(c)1-37) or 38-63 (F(c)38-63) of AE1-4 accumulated in intracellular membrane compartments that overlapped late endosomes and the trans-Golgi network (TGN), respectively. Internalization assays indicated that the patterns of localization observed for F(c)1-37 and F(c)38-63 resulted from the recycling of these chimeras from the cell surface. These assays further indicated that F(c)1-37 and F(c)38-63 each possess a basolateral sorting activity. Mutagenesis studies revealed that the endocytic and basolateral sorting activities in F(c)1-37 are dependent upon serine 25, which is located in a sequence similar to a sorting signal in the polymeric immunoglobulin receptor. In addition, the sorting activities associated with F(c)38-63 were dependent upon tyrosine 47 and leucine 50. These residues resided within the sequence, YVEL, which matches the YXXPhi motif (where X is any amino acid and Phi is a hydrophobic residue) that functions as an endocytic and TGN recycling signal for other membrane proteins. Our data indicate that amino acids 1-63 of AE1-4 contain sorting and cytoskeletal binding activities that account for most of the properties previously associated with AE1-4 in MDCK cells. Furthermore, the alternative localization patterns exhibited by chimeras containing various combinations of these activities suggest that interplay between these cytoplasmic activities is critical for specifying AE1-4 localization in epithelial cells. PMID- 12538767 TI - VEGF expression in human macrophages is NF-kappaB-dependent: studies using adenoviruses expressing the endogenous NF-kappaB inhibitor IkappaBalpha and a kinase-defective form of the IkappaB kinase 2. AB - Vascular endothelial growth factor (VEGF) is the most endothelial cell-specific angiogenic factor characterised to date, and it is produced by a variety of cell types. In macrophages, VEGF has been shown to be upregulated by the inflammatory mediator lipopolysaccharide (LPS) and by engagement of CD40 by CD40 ligand (CD40L). Because LPS and CD40L activate nuclear factor-kappaB (NF-kappaB) in monocytes, we investigated in this study whether VEGF production in macrophages, when stimulated with either LPS or CD40L, is NF-kappaB-dependent. We used adenoviral constructs over-expressing either IkappaBalpha (AdvIkappaBalpha), the endogenous inhibitor of NF-kappaB, or a kinase-defective mutant of IKK-2 (AdvIKK 2dn), an upstream activator of IkappaBalpha, to infect normal human monocyte derived macrophages. We observed that LPS-induced production of VEGF in human macrophages was almost completely inhibited (>90%) following adenoviral transfer of IkappaBalpha. In addition, we observed significant inhibition of the CD40L induced VEGF production in macrophages following infection with AdvIkappaBalpha. Expression of IKK-2dn in macrophages decreased VEGF production in response to LPS or CD40L by approximately 50%, suggesting that in addition to IKK-2, other kinases might be involved in NF-kappaB activation. These results show for the first time that VEGF production in human macrophages is NF-kappaB dependent. NF kappaB regulates many of the genes involved in immune and inflammatory responses, and our study adds the angiogenic cytokine VEGF to the list of NF-kappaB dependent cytokines. PMID- 12538768 TI - Insulin-like growth factor binding protein 5 and apoptosis in mammary epithelial cells. AB - Insulin-like growth factors (IGFs) are important survival signals that can protect a range of cell types from apoptosis. Although IGF bioavailability is modulated by high affinity interactions with IGF-binding proteins (IGFBPs), there is currently no experimental evidence that IGFBPs regulate the survival function of IGFs in the mammary gland. We have examined IGFBP expression during mammary gland development and studied the effects of IGFBPs on IGF-mediated survival and signalling in mammary epithelial cells in culture. IGFBP-5 protein was greatly increased during days 1-3 of mammary gland involution, when levels of apoptosis are dramatically elevated to remodel the gland after lactation. Primary cultures of mammary epithelial cells (MECs) expressed IGFBP-5 from their basal surface suggesting that IGFBP-5 is suitably located to inhibit IGF signalling. Addition of exogenous IGFBP-5 and IGFBP-3 to MECs suppressed IGF-I-mediated survival, resulting in threefold greater apoptosis in cells incubated with IGF-I and IGFBP 5 compared with IGF-I alone. Examination of signalling pathways involved in apoptosis revealed that phosphorylation of PKB and the forkhead transcription factor, FKHRL1, was induced by IGFs, but that phosphorylation was blocked by IGFBP-5 and IGFBP-3. This study provides evidence that IGFBP-5 plays an important role in the regulation of apoptosis in the mammary gland. PMID- 12538769 TI - Deletion of exon 4 from human surfactant protein C results in aggresome formation and generation of a dominant negative. AB - Human surfactant protein C (hSP-C) is synthesized by the alveolar type 2 cell as a 197 amino acid integral membrane proprotein and proteolytically processed to a secreted 3.7 kDa mature form. Although the SP-C null mouse possesses a non-lethal phenotype, a heterozygous substitution of A for G in the first base of intron 4 of the human SP-C gene (c.460+1A>G) has been reported in association with familial interstitial lung disease and absence of mature protein. This mutation produces a splice deletion of exon 4 (deltaExon4) resulting in removal of a positionally conserved cysteine in the C-terminal flanking propeptide. Based on a prior study showing that an identical deletion in the rat isoform diverted mutant protein to stable aggregates, we hypothesized that expression of the deltaExon4 mutation would result in disruption of intracellular trafficking of both mutant and wild-type proSP-C. We tested this in vitro using fusion proteins of EGFP conjugated either to wild-type SP-C (EGFP/hSP-C(1-197)) or to SP-C deleted of Exon4 (EGFP/hSP-C(deltaExon4)). Fluorescence microscopy showed that EGFP/hSP-C(1 197) transfected into A549 cells was expressed in a punctuate pattern in CD63 (+) cytoplasmic vesicles, whereas EGFP/hSP-C(deltaExon4) accumulated in ubiquitinated perinuclear inclusions linked to the microtubule organizing center. A similar juxtanuclear pattern was observed following transfection of SP-C cDNA lacking only cysteine residues in the C-terminal propeptide encoded by Exon 4 (EGFP/hSP C(C120/121G)). To evaluate whether mutant proSP-C could function as a dominant negative, EGFP/hSP-C(deltaExon4) was cotransfected with HA-tagged hSP-C(1-197) and resulted in the restriction of both forms to perinuclear compartments. Addition of Na(+) 4-phenylbutyrate, a facilitator of trafficking of other misfolded proteins, attenuated the aggregation of EGFP/hSP-C(deltaExon4). We conclude that c.460+1A>G mutation of human SP-C results in disruption of disulfide-mediated folding encoded by Exon 4 leading to diversion of unprocessed proSP-C to aggresomes. The heterotypic oligomerization of hSP-C(1-197) and hSP C(deltaExon4) provides a molecular mechanism for the dominant-negative effect observed in vivo. PMID- 12538770 TI - Protection against hypoxia-induced increase in blood-brain barrier permeability: role of tight junction proteins and NFkappaB. AB - Co-culture with glial cells and glia-conditioned media can induce blood-brain barrier properties in microvessel endothelial cells and protect against hypoxia induced blood-brain barrier breakdown. We examined the effect of two types of glia-conditioned media on brain microvessel endothelial cell permeability and tight junction protein expression, and studied potential mechanisms of action. We found that C6-glioma-conditioned media, but not rat astrocyte-conditioned media, protected against an increase in permeability induced by exposure to 1% oxygen for 24 hours. This hypoxic stress caused an increase in the expression of tight junction proteins claudin-1 and actin, particularly in cells treated with C6 conditioned media. We found that C6-conditioned media has a significantly higher level of both basic fibroblast growth factor and vascular endothelial growth factor. Treatment with C6-conditioned media for 1 or 3 days protects against hypoxia-induced permeability increases, and this protective effect may be mediated by signal transduction pathways terminating at the transcription factor NFkappaB. PMID- 12538771 TI - Krh1p and Krh2p act downstream of the Gpa2p G(alpha) subunit to negatively regulate haploid invasive growth. AB - The yeast G(alpha) subunit Gpa2p and its coupled receptor Gpr1p function in a signaling pathway that is required for the transition to pseudohyphal and invasive growth. A two-hybrid screen using a constitutively active allele of GPA2 identified the KRH1 gene as encoding a potential binding partner of Gpa2p. Strains containing deletions of KRH1 and its homolog KRH2 were hyper-invasive and displayed a high level of expression of FLO11, a gene involved in pseudohyphal and invasive growth. Therefore, KRH1 and KRH2 encode negative regulators of the invasive growth pathway. Cells containing krh1Delta krh2Delta mutations also displayed increased sensitivity to heat shock and decreased sporulation efficiency, indicating that Krh1p and Krh2p regulate multiple processes controlled by the cAMP/PKA pathway. The krh1Delta krh2Delta mutations suppressed the effect of a gpa2Delta mutation on FLO11 expression and eliminated the effect of a constitutively active GPA2 allele on induction of FLO11 and heat shock sensitivity, suggesting that Krh1p and Krh2p act downstream of Gpa2p. The Sch9p kinase was not required for the signal generated by deletion of KRH1 and KRH2; however, the cAMP-dependent kinase Tpk2p was required for generation of this signal. These results support a model in which activation of Gpa2p relieves the inhibition exerted by Krh1p and Krh2p on components of the cAMP/PKA signaling pathway. PMID- 12538772 TI - ForC, a novel type of formin family protein lacking an FH1 domain, is involved in multicellular development in Dictyostelium discoideum. AB - Formins are highly conserved regulators of cytoskeletal organization and share three regions of homology: the FH1, FH2 and FH3 domains. Of the nine known formin genes or pseudogenes carried by Dictyostelium, forC is novel in that it lacks an FH1 domain. Mutant Dictyostelium lacking forC (DeltaforC) grew normally during the vegetative phase and, when starved, migrated normally and formed tight aggregates. Subsequently, however, DeltaforC cells made aberrant fruiting bodies with short stalks and sori that remained unlifted. DeltaforC aggregates were also unable to migrate as slugs, suggesting forC is involved in mediating cell movement during multicellular stages of Dictyostelium development. Consistent with this idea, expression of forC was increased significantly in aggregates of wild-type cells. GFP-ForC expressed in DeltaforC cells was localized at the crowns, which are macropinocytotic structures rich in F-actin, suggesting that, like other formin isoforms, ForC functions in close relation with the actin cytoskeleton. Truncation analysis of GFP-ForC revealed that the FH3 domain is required for ForC localization; moreover, localization of a truncated GFP-ForC mutant at the site of contacts between cells on substrates and along the cortex of cells within a multicellular culminant suggests that ForC is involved in the local actin cytoskeletal reorganization mediating cell-cell adhesion. PMID- 12538773 TI - Constitutive activation of Rho proteins by CNF-1 influences tight junction structure and epithelial barrier function. AB - The apical-most epithelial intercellular junction, referred to as the tight junction (TJ), regulates paracellular solute flux in diverse physiological and pathological states. TJ affiliations with the apical filamentous actin (F-actin) cytoskeleton are crucial in regulating TJ function. F-actin organization is influenced by the Rho GTPase family, which also controls TJ function. To explore the role of Rho GTPases in regulating TJ structure and function, we utilized Escherichia coli cytotoxic necrotizing factor-1 (CNF-1) as a tool to activate constitutively Rho, Rac and Cdc42 signaling in T84 polarized intestinal epithelial monolayers. The biological effects of the toxin were polarized to the basolateral membrane, and included profound reductions in TJ gate function, accompanied by displacement of the TJ proteins occludin and zonula occludens-1 (ZO-1), and reorganization of junction adhesion molecule-1 (JAM-1) away from the TJ membrane. Immunogold electron microscopy revealed occludin and caveolin-1 internalization in endosomal/caveolar-like structures in CNF-treated cells. Immunofluorescence/confocal microscopy suggested that a pool of internalized occludin went to caveolae, early endosomes and recycling endosomes, but not to late endosomes. This provides a novel mechanism potentially allowing occludin to evade a degradative pathway, perhaps allowing efficient recycling back to the TJ membrane. In contrast to the TJ, the characteristic ring structure of proteins in adherens junctions (AJs) was largely preserved despite CNF-1 treatment. CNF-1 also induced displacement of a TJ-associated pool of phosphorylated myosin light chain (p-MLC), which is normally also linked to the F-actin contractile machinery in epithelial cells. The apical perjunctional F-actin ring itself was maintained even after toxin exposure, but there was a striking effacement of microvillous F actin and its binding protein, villin, from the same plane. However, basal F actin stress fibers became prominent and cabled following basolateral CNF-1 treatment, and the focal adhesion protein paxillin was tyrosine phosphorylated. This indicates differences in Rho GTPase-mediated control of distinct F-actin pools in polarized cells. Functionally, CNF-1 profoundly impaired TJ/AJ assembly in calcium switch assays. Re-localization of occludin but not E-cadherin along the lateral membrane during junctional reassembly was severely impaired by the toxin. A balance between activity and quiescence of Rho GTPases appears crucial for both the generation and maintenance of optimal epithelial barrier function. Overactivation of Rho, Rac and Cdc42 with CNF-1 seems to mirror key barrier function disruptions previously reported for inactivation of RhoA. PMID- 12538774 TI - Microtubule involvement in NIH 3T3 Golgi and MTOC polarity establishment. AB - Scratch-wound assays are commonly used to study the ability of cells to polarize and migrate. In a previous study we showed that Golgi reorientation in response to a scratch wound is actin-dependent in NIH 3T3 cells but not in astrocytes. In this investigation, to study cell polarity and motility further, we used the polarization of the Golgi and microtubule organizing center (MTOC), as well as the ability of NIH 3T3 cells to migrate, in a scratch-wound assay. Unlike Golgi polarization, MTOC polarization was not dependent on actin, the Arp2/3 complex or Wiskott-Aldrich syndrome protein (WASP)-family proteins. By contrast, disruption of microtubules inhibited MTOC polarity, but not Golgi polarity. Migration was found to be dependent both on actin and microtubules. Expression of the formin homology 2 (FH2) region of mDia1 inhibited Golgi polarization and migration but not MTOC polarization. Similarly, ST638, a Src inhibitor, inhibited Golgi polarization and migration but not MTOC polarization, whereas expression of the actin regulator IRSp53 only inhibited cell migration. Interestingly, the inhibition of cell migration by the mDia1 FH2 domain could be overcome by addition of Y27632, an inhibitor of ROCK (Rho-associated kinase). In fact, in the presence of ROCK inhibitor, cell migration was accelerated but polarization of both the Golgi and MTOC were inhibited. These data show that, in NIH 3T3 cells, different aspects of cell polarization and migration occur by different mechanisms, and both actin and microtubule networks are required. In addition, this study indicates that MTOC and Golgi polarization events are separately controlled. PMID- 12538775 TI - Vernix, the newborn, and innate defense. PMID- 12538776 TI - Delayed puberty associated with inflammatory bowel disease. AB - Delayed puberty frequently complicates the clinical course of young patients with inflammatory bowel disease, more often in Crohn's disease than ulcerative colitis. Undernutrition has been thought to be the main reason for delayed puberty in these patients. However, puberty may be delayed despite a normal nutritional status. Observations in patients with inflammatory bowel disease and in rats with experimental colitis suggest that inflammatory mediators may have a direct adverse influence, independent of undernutrition, on the onset and progression of puberty. Serum androgens are consistently reported to be reduced in patients with delayed puberty and inflammatory bowel disease. This reduction is not necessarily secondary to a reduction in gonadotrophins as serum concentrations of gonadotrophins have been reported to be normal or even increased in some studies. Management of delayed puberty involves calorie supplements to correct undernutrition and treatment of inflammation. Observations in boys with delayed puberty and controlled studies in experimental models of intestinal inflammation suggest that testosterone therapy can accelerate puberty. PMID- 12538777 TI - Antimicrobial polypeptides of human vernix caseosa and amniotic fluid: implications for newborn innate defense. AB - Antimicrobial peptides/proteins are widespread in nature and play a critical role in host defense. To investigate whether these components contribute to surface protection of newborns at birth, we have characterized antimicrobial polypeptides in vernix caseosa (vernix) and amniotic fluid (AF). Concentrated peptide/protein extracts were obtained from 11 samples of vernix and six samples of AF and analyzed for antimicrobial activity using an inhibition zone assay. Proteins/peptides in all vernix extracts exhibited strong antibacterial activity against Bacillus megaterium (strain Bm11), in addition to antifungal activity against Candida albicans, whereas AF-derived proteins/peptides showed only the former activity. Fractions obtained after separation by reverse-phase HPLC exhibited antibacterial activity, with the most pronounced activity in a fraction containing alpha-defensins (HNP1-3). The presence of HNP1-3 was proved by dot blot analysis and confirmed by mass spectrometry. Lysozyme and ubiquitin were identified by sequence analysis in two fractions with antibacterial activity. Fractions of vernix and AF were also positive for LL-37 with dot blot and Western blot analyses, and one fraction apparently contained an extended form of LL-37. Interestingly, psoriasin, a calcium-binding protein that is up-regulated in psoriatic skin and was found recently to exhibit antimicrobial activity, was characterized in the vernix extract. The presence of all of these antimicrobial polypeptides in vernix suggests that they are important for surface defense and may have an active biologic role against microbial invasion at birth. PMID- 12538778 TI - Iron deficiency anemia in infancy: long-lasting effects on auditory and visual system functioning. AB - Evoked potentials provide noninvasive measures of nerve transmission and CNS functioning. Auditory brainstem responses (ABR) and visual evoked potentials (VEP) show dramatic changes in infancy, largely as a result of progressive myelination. Because iron is required for normal myelination, pathway transmission in these sensory systems might be affected by early iron deficiency. We previously reported evidence to that effect: infants with iron-deficiency anemia (IDA) had slower transmission through the auditory brainstem pathway, uncorrected by iron therapy. To determine long-term effects, ABR and/or VEP of healthy Chilean children who were treated for IDA or were nonanemic in infancy were compared at approximately 4 y of age. Absolute latencies for all ABR waves and interpeak latencies (except I-III interval) were significantly longer in former IDA children. Longer latency was also observed for the P100 wave on VEP. The magnitude of differences was large-about 1 SD. These findings, with differences in latencies but not amplitudes, further support the hypothesis that IDA in infancy alters myelination and provide evidence that effects on transmission through the auditory and visual systems can be long lasting. Subtle changes in sensory pathway transmission might be an underlying mechanism for the derailment of other developmental aspects in early IDA. PMID- 12538779 TI - Mutation screening in patients with isolated cytochrome c oxidase deficiency. AB - Cytochrome c oxidase (COX) deficiency has been associated with a variety of clinical conditions and can be due to mutations in nuclear or mitochondrial genes. Despite recent progress in our understanding of the molecular bases of COX deficiency, the genetic defect remains elusive in many cases. We performed mutation screening in 30 patients with biochemical evidence of isolated COX deficiency and heterogeneous clinical phenotypes. Sixteen patients had various forms of encephalomyopathy, and six of these had the neuroradiological features of Leigh syndrome. Four patients had encephalohepatopathy, six had hypertrophic cardiomyopathy, and four had other phenotypes. We studied the three mtDNA genes encoding COX subunits, the 22 mtDNA tRNA genes, and seven COX assembly genes: SCO1, SCO2, SURF1, COX10, COX11, COX15, and COX17. We report two novel pathogenic SURF1 mutations in a patient with Leigh syndrome and one novel SCO2 mutation in a patient with hypertrophic cardiomyopathy. These data show that heterogeneous clinical phenotypes are associated with COX deficiency, that mutations in mtDNA COX genes are rare, and that mutations in additional genes remain to be identified. PMID- 12538780 TI - Dexamethasone potentiates keratinocyte growth factor-stimulated SP-A and SP-B gene expression in alveolar epithelial cells. AB - Keratinocyte growth factor (KGF, or fibroblast growth factor 7) was previously reported to enhance the synthesis of surfactant in alveolar type II cells. We investigated the possible interactions between KGF and a glucocorticoid, dexamethasone (Dex), on surfactant protein (SP) gene expression. In cultured fetal rat type II cells, KGF and Dex induced greater-than-additive stimulating effects on SP-A and SP-B expressions that were enhanced three-fold and 30-fold, respectively, but had only additive effects on SP-C expression. Using murine lung epithelial (MLE) cells, KGF increased SP-A, SP-B (up to two-fold), and SP-C (up to three-fold) mRNA levels in a dose-dependent way. Dex 10(-9) to 10(-7) M increased SP-A and SP-B mRNA 1.5-fold and SP-C mRNA two-fold. Consistent with type II cell findings, simultaneous treatment by KGF and Dex induced a synergistic increase of SP-A and SP-B transcripts (three-fold and 4.5-fold, respectively), but not of SP-C transcripts. SP-A protein was present in MLE-15 and was increased about three-fold by KGF plus Dex. Expression study of a reporter gene placed under either the SP-A or the SP-B gene regulatory sequences and transfected in MLE-15 cells indicated that the Dex-KGF synergy was achieved mainly through a transcriptional effect for SP-A, and both transcriptional and nontranscriptional effects for SP-B. For the latter, increased mRNA stability was evidenced with the aid of actinomycin D. The Dex-KGF synergy may have potential interest for diseases associated with surfactant deficiency. PMID- 12538781 TI - Peptide growth factors in tracheal aspirates of mechanically ventilated preterm neonates. AB - Basic fibroblast growth factor (bFGF or FGF-2), vascular endothelial growth factor (VEGF), and endothelin-1 (ET-1) are peptide growth factors (PGF) mediating normal lung development, maturation, injury, and repair. These PGF may therefore be involved in the pathogenesis of bronchopulmonary dysplasia (BPD). We hypothesized that elevated levels of these PGF in tracheal aspirates would be associated with a) BPD and/or death; b) markers of cell injury and apoptosis; and c) chorioamnionitis, a risk factor for BPD. Tracheal aspirates collected in 29 preterm (<34 wk gestation, 500-2000 g birth weight), mechanically ventilated infants on d 1 of life were assayed for PGF and histone-associated DNA fragments by ELISA and for LDH by enzyme assay. Clinical and pathologic examination was performed for chorioamnionitis. BPD was defined as oxygen requirement/mechanical ventilation at 28 d postnatal age. The birth weight (mean +/- SE) was 1009 +/- 85 g and median gestational age was 26 wk (range, 22-33). Eighteen infants died or developed BPD. bFGF levels were elevated in infants who died or developed BPD [median (25%,75%) level of 36 (23, 44) pg/mL versus 14 (6, 30) in the survivors without BPD, p = 0.01]. bFGF levels correlated with apoptosis (r = 0.73, p < 0.001) and LDH levels (r = 0.59, p < 0.001). VEGF and ET-1 levels were not associated with apoptosis or with BPD/death. PGF levels were not associated with chorioamnionitis. We conclude that elevated bFGF levels in the preterm trachea correlate with BPD/death and markers of cell injury and apoptosis but not with chorioamnionitis. We speculate that bFGF may play a role in the development of BPD. PMID- 12538782 TI - Positive pressure ventilation with the open lung concept optimizes gas exchange and reduces ventilator-induced lung injury in newborn piglets. AB - Previous studies demonstrated that high-frequency oscillatory ventilation using the open lung concept (OLC) resulted in superior gas exchange and a reduction in ventilator-induced lung injury (VILI). We hypothesized that these beneficial effects could also be achieved by applying the OLC during positive pressure ventilation. After repeated whole-lung-lavage, newborn piglets were assigned to either OLC positive pressure ventilation (PPV(OLC)), OLC high-frequency oscillatory ventilation (HFOV(OLC)), or conventional positive pressure ventilation (PPV(CON)) and ventilated for 5 h. In both OLC groups, collapsed alveoli were actively recruited and thereafter stabilized using the lowest possible airway pressures. In the PPV(CON) group, ventilator settings were adjusted to prevent critical hypoxia. Airway pressure, blood gas analysis, pressure-volume curve, and alveolar protein infiltration was recorded. A lung injury score was used for histologic comparison. Mean airway pressures were comparable in the three ventilation groups over time (1.2-1.5 kPa). Arterial oxygenation increased to mean values above 60 kPa in both OLC groups compared with 10 kPa in the PPV(CON) group (p < 0.001). Maximal lung compliance was superior in both OLC groups (PPV(OLC): 91 +/- 23; HFOV(OLC): 90 +/- 31 mL/kPa/kg, p < 0.01) compared with the PPV(CON) group (39 +/- 14 mL/kPa/kg). Alveolar protein infiltration was significantly reduced in the PPV(OLC) group (0.33 +/- 0.10 mg/mL, p < 0.01) and the HFOV(OLC) group (0.40 +/- 0.13 mg/mL, p < 0.01) compared with the PPV(CON) group (0.70 +/- 0.15 mg/mL). Lung injury scores were significantly higher in the PPV(CON) group (33.5 +/- 9.5, p < 0.01) compared with both OLC groups (PPV(OLC): 10.5 +/- 2.6; HFOV(OLC): 11 +/- 2.2). There were no differences between the two OLC groups. We conclude that, in surfactant-depleted newborn piglets, application of the OLC during PPV is feasible and results in superior gas exchange and a reduction in VILI compared with conventional PPV. These beneficial effects are comparable to HFOV. PMID- 12538783 TI - Lung epithelial cells undergo apoptosis in neonatal respiratory distress syndrome. AB - For studying the presence of programmed cell death in the lungs of infants with fatal respiratory distress syndrome (RDS) and the possible contribution of postnatal glucocorticoid administration on this cell destruction, lung tissue samples from autopsies of 16 premature infants with fatal RDS were studied. The infants had neither been exposed to antenatal steroids nor received surfactant therapy, but seven of these infants had been subjected to postnatal dexamethasone treatment. Lung autopsy samples of seven term and two preterm neonates without any obvious lung disease served as controls. Lungs were studied histologically, and apoptotic cell death was identified using DNA nick end-labeling assay and caspase-related M30 antibody staining (CytoDeath). Lung tissue from the RDS infants showed elevated leukocyte infiltration, histologic injury score, and number of apoptotic cells, located mainly in the respiratory epithelium, when compared with controls. In contrast, lungs from infants who had RDS and received dexamethasone demonstrated markedly reduced tissue leukocyte accumulation and injury score and lower rates of epithelial apoptosis than the lungs of infants who had RDS and did not receive dexamethasone. These results suggest that significant epithelial apoptosis is present in the lungs of newborn infants with fatal RDS and that this apoptosis may be attenuated by steroid administration. PMID- 12538784 TI - Dexamethasone pretreatment attenuates cerebral vasodilative responses to hypercapnia and augments vasoconstrictive responses to hyperventilation in newborn pigs. AB - In the perinatal period, glucocorticoids are frequently administered to enhance pulmonary maturity or prevent chronic lung disease of prematurity. Recently, it has been suggested that the perinatal exposure to glucocorticoids can be associated with unfavorable neurologic development. We studied the hypothesis that 24-h pretreatment with glucocorticoid might modify cerebrovascular responses to high and low partial arterial CO(2) tension in newborn animals in vivo. A closed cranial window was implanted over the left parietal cortex of 20 anesthetized ventilated newborn (<3 d old) pigs. The actual experiments were carried out in 15 pigs: eight pretreated with a total dose of 6 mg/kg of dexamethasone and seven controls. Five pigs were used for preliminary experiments as described in the text. Pial arteriolar diameters were measured during 1) baseline conditions (normocapnia), 2) hypercapnia induced by ventilating the animals with a gas mixture containing 10% CO(2), or 3) hyperventilation with resultant hypocapnia. Under these conditions, the concentrations of 6-keto PGF(1alpha) in the CSF were measured in five experimental animals and six controls. In summary, the dexamethasone pretreatment 1) attenuated the hypercapnia-induced dilator responses of pial arterioles and prevented the hypercapnia-associated fall in mean arterial blood pressure; 2) caused moderate, although not statistically significant, diminution in 6-keto-PGF(1alpha) levels in the CSF during baseline; 3) blocked hypercapnia-induced elevation of 6-keto PGF(1alpha); and 4) enhanced vasoconstrictive arteriolar responses to hyperventilation. We speculate that in the clinical setting, the dexamethasone effects may compromise the adjustments of global or regional cerebral blood flow to changing physiologic states in neonates. PMID- 12538785 TI - Consequences of in utero caffeine exposure on respiratory output in normoxic and hypoxic conditions and related changes of Fos expression: a study on brainstem spinal cord preparations isolated from newborn rats. AB - Several aspects of the central regulation of respiratory control have been investigated on brainstem-spinal cord preparations isolated from newborn rats whose dam was given 0.02% caffeine in water as drinking fluid during the whole period of pregnancy. Analysis of the central respiratory drive estimated by the recording of C4 ventral root activity was correlated to Fos ponto-medullary expression. Under normoxic conditions, preparations obtained from the caffeine treated group of animals displayed a higher respiratory frequency than observed in the control group (9.2 +/- 0.5 versus 7.2 +/- 0.6 burst/min). A parallel Fos detection tends to indicate that the changes of the respiratory rhythm may be due to a decrease in neuronal activity of medullary structures such as the ventrolateral subdivision of the solitary tract, the area postrema, and the nucleus raphe obscurus. Under hypoxic conditions, the preparations displayed a typical hypoxic respiratory depression associated with changes in the medullary Fos expression pattern. In addition, the hypoxic respiratory depression is clearly emphasized after in utero exposure to caffeine and coincides with an increased Fos expression in the area postrema and nucleus raphe obscurus, two structures in which it is not increased in the absence of caffeine. Taken together, these results support the idea that in utero caffeine exposure could affect central respiratory control. PMID- 12538786 TI - Gene expression and activity of enzymes in the arginine biosynthetic pathway in porcine fetal small intestine. AB - We recently reported the presence of ornithine aminotransferase (OAT) enzymatic activity and mRNA expression in the intestine of fetal pigs from 30 to 110 d of gestation. Here we describe the activities and mRNA expression patterns of other key enzymes in the arginine biosynthetic pathway, specifically carbamoyl phosphate synthase I (CPS-I), ornithine carbamoyl transferase (OCT), and pyrroline-5-carboxylate reductase (P5CR), in the fetal porcine small intestine from 30 to 110 d of gestation. The activities of all three enzymes increased from d 30 to d 110 of gestation, and in situ hybridization demonstrates that 1) CPS-I and OCT genes are expressed in distinct patterns and are confined to the mucosal epithelium and 2) P5CR mRNA is present in mucosal epithelium and lamina propria of the fetal porcine small intestine from d 30 to d 110 of gestation. The presence of CPS-I and OCT in conjunction with the presence of OAT suggests that the fetal porcine small intestine is capable of synthesizing citrulline from P5C. In addition, the presence of P5CR suggests that the fetal porcine small intestine is able to synthesize proline from ornithine via OAT. This ability of the fetal small intestine to synthesize amino acids may be important for development and metabolic activity of the intestine during somatic growth of the fetus. PMID- 12538787 TI - Effect of minimal enteral feeding on splanchnic uptake of leucine in the postabsorptive state in preterm infants. AB - We conducted a controlled, randomized trial to study the effect of minimal enteral feeding on leucine uptake by splanchnic tissues, as an indicator of maturation of these tissues, in preterm infants in the first week of life. Within a few hours after birth, while receiving only glucose, a primed constant infusion of [1-(13)C]-leucine was started and continued for 5 h via the nasogastric tube, whereas 5,5,5 D3-leucine was infused intravenously (for both tracers, priming dose 2 mg/kg, continuous infusion 2 mg/kg/h). Patients were thereafter randomized to receive solely parenteral nutrition (C), parenteral nutrition and 20 mL breast milk/kg/d (BM), or parenteral nutrition and 20 mL formula/kg/d (F). On d 7, the measurements were repeated, after discontinuing the oral intake for 5 h. Fourteen infants were included in group C, 12 in group BM, and 12 in group F. There was no difference in energy intake or nitrogen balance at any time. On d 1, plasma enrichment for the nasogastric tracer was lower than for the intravenous tracer for all three groups, both for leucine and for alpha-keto-isocaproic acid. On d 7, the enrichment for leucine and alpha-keto-isocaproic acid for the nasogastric tracer was lower than for the intravenous tracer for the groups BM and F (BM: 3.65 +/- 1.20 nasogastric versus 4.64 +/- 0.64 i.v.; F: 4.37 +/- 1.14 nasogastric versus 5.21 +/- 0.9 i.v.). In the control group, there was no difference between tracers. The lower plasma enrichment for the nasogastric tracer compared with the intravenous tracer suggests uptake of leucine by the splanchnic tissues. We conclude that minimal enteral feeding--even in low volumes of 20 mL/kg/d- increases the leucine uptake by the splanchnic tissue. We speculate that this reflects a higher protein synthesis of splanchnic tissues in the groups receiving enteral nutrition. PMID- 12538788 TI - Differential developmental regulation of rat liver canalicular membrane transporters Bsep and Mrp2. AB - Bile formation depends on the active secretion of bile salts and other biliary constituents by specific transporters. Recently two major transporters that contribute to bile formation, the bile salt export pump (Bsep) and multidrug resistance protein-2 (Mrp2), have been cloned. The goal of the present study was to define the expression of Bsep and Mrp2 during rat liver development. mRNA expression as assessed by Northern blot and RT-PCR was higher for Mrp2 (40% of adult) at 21 d fetal age relative to Bsep (<20% of adult). The levels of Mrp2 mRNA increased to approximately 50% of adult at 1 d of life and then rapidly increased to adult levels by 1-3 wk. Nuclear run-on assays for Bsep and Mrp2 showed minimal transcription during fetal life with an increase in transcription in the postnatal period. A different pattern of expression was observed for both Mrp2 and Bsep proteins. During fetal life, there was low expression of Mrp2 and Bsep proteins (<20% of adult) with a gradual increase neonatally reaching adult levels at 4 wk. Thus, we noted a temporal delay between the maximal expression of the mRNA (1-3 wk) and protein (4 wk) for Bsep and Mrp2. These results show that 1) expression (of mRNA and protein) of canalicular transporters is developmentally regulated by both transcriptional and posttranscriptional mechanisms and 2) Mrp2 and Bsep gene expression (mRNA) are differentially regulated. PMID- 12538789 TI - Distribution of the IgG Fc receptor, FcRn, in the human fetal intestine. AB - The intestinal Fc receptor, FcRn, functions in the maternofetal transfer of gamma globulin (IgG) in the neonatal rodent. In humans, most of this transfer is presumed to occur in utero via the placenta. Although the fetus swallows amniotic fluid that contains immunoglobulin, it is unknown whether this transfer also occurs via the fetal intestine. A human FcRn has been identified in the syncytiotrophoblast that mediates the maternofetal transfer of antibody. It has also been identified in human fetal intestine and is postulated to function in IgG transport. We hypothesize that the human fetal intestinal FcRn may play a role in IgG transport from the amniotic fluid into the fetal circulation. The aim of this study was to characterize the distribution of the FcRn along the human fetal intestine. Lysates prepared from human fetal intestine and from a nonmalignant human fetal intestinal epithelial cell line (H4) were subjected to Western blot analysis and probed using anti-FcRn antibodies. A 42-kD band, consistent with the known molecular weight of the FcRn, was detected along the human fetal intestine and in H4 cells. Expression of the human FcRn was confirmed with immunohistochemistry. Our study demonstrates the expression of FcRn along the human fetal intestine and in a human nonmalignant fetal intestinal epithelial cell line (H4), which by location indicates that FcRn could play a role in the uptake and transport of IgG in the human fetus. PMID- 12538790 TI - Differential effects of fetal number and maternal nutrition in late gestation on prolactin receptor abundance and adipose tissue development in the neonatal lamb. AB - The present study examined the extent to which abundance of the prolactin receptor (PRLR) and a range of primary mitochondrial proteins are influenced by either maternal nutrition and/or fetal number in adipose tissue. Pregnant sheep were control fed [consuming 100% of total metabolizable energy (ME) requirements (taking into account requirements for both ewe maintenance and growth of the conceptus to produce a 4.5-kg lamb at term) for that stage of gestation] or were nutrient restricted (consuming 60% of total ME requirements). All ewes lambed normally at term and both perirenal adipose and hepatic tissues were sampled within 6 h of birth. Plasma membranes and mitochondria were prepared and analyzed using immunoblotting for abundance of PRLR and/or cytochrome c, voltage-dependent anion channel (VDAC), and uncoupling protein-1 (UCP1). Irrespective of maternal nutrition, abundance of specific isoforms of PRLR were significantly higher in adipose tissue sampled from twins compared with singletons and total UCP1 concentration per milligram of tissue was increased (p < 0.05). There was no effect of fetal number on PRLR abundance in the liver. Maternal nutrient restriction resulted in an increased abundance of both cytochrome c (p < 0.001) and VDAC in adipose tissue of twins but not singletons. This occurred despite maternal nutrition having no effect on either lamb body or adipose tissue weights, although both were lower (p < 0.05) in twins compared with singletons. In conclusion, fetal adipose tissue development is highly sensitive to nutritionally mediated changes in late gestation. An increase in fetal number results in greater PRLR abundance, which, in conjunction with a decrease in maternal nutrition, results in up-regulation of specific mitochondrial proteins. PMID- 12538791 TI - CD8+ T-cell numbers predict the response to antiviral therapy in HIV-1-infected children. AB - Our objective was to study the probability of achieving undetectable viral load levels in HIV-1-infected children after 36 mo of highly active antiretroviral therapy (HAART). A prospective, multicenter, longitudinal study in 41 HIV-1 infected children on HAART was undertaken. Viral load was quantified using standard molecular assay. CD4+ and CD8+ T cell subsets were determined by flow cytometry. The probability of achieving undetectable viral load was determined using Kaplan-Meier curves according to groups by percentage CD8+ at baseline (CD8+ <25% or >25%). Lower baseline CD8+ T cell levels conditioned a less effective virological response to HAART in children, independent of baseline CD4+ T cell numbers and viral load levels. A greater number of children (81%) from CD8+ >25% group than from the CD8+ <25% (40%) presented undetectable viral load levels (p = 0.013). Additionally, the CD8+ >25% group showed a 4.5-fold higher (95% confidence interval: 1.1-19.2) relative proportion for achieving viral load <400 copies/mL than the CD8+ <25% group (p = 0.039). We concluded that monitoring CD8+ T cell numbers may be valuable in deciding when to start HAART in vertically HIV-1-infected children. PMID- 12538792 TI - IL-10 controls Aspergillus fumigatus- and Pseudomonas aeruginosa-specific T-cell response in cystic fibrosis. AB - Up to 90% of patients with cystic fibrosis (CF) are chronically colonized with Pseudomonas aeruginosa, and 10% to 50% of CF patients are colonized with Aspergillus fumigatus. Despite an extensive inflammatory reaction, patients cannot eliminate the microorganisms. The present study demonstrates that an IL-10 mediated T-cell tolerance to major infectious agents A. fumigatus and P. aeruginosa plays an important role in the control of T-cell-mediated inflammatory responses in CF. Peripheral blood mononuclear cells of CF patients secreted significantly higher amounts of IL-10. T-cell response against recombinant A. fumigatus antigens rAsp f 3, rAsp f 4, rAsp f 6, and heat-inactivated P. aeruginosa was controlled by IL-10. Proliferation and interferon-gamma production was significantly increased when endogenous IL-10 was blocked in aspergillus and pseudomonas antigen-stimulated cells of CF patients. The role of IL-10 was further documented by increased spontaneous proliferation of peripheral blood mononuclear cells of CF patients after preincubation with antisense oligonucleotides blocking the synthesis of IL-10 receptor-associated kinases janus tyrosine kinase 1 and tyrosine kinase 2. Together, these data demonstrate an important role of IL-10-mediated peripheral T-cell tolerance to P. aeruginosa and A. fumigatus in the control of the intensity of the inflammatory T-cell response in CF. PMID- 12538793 TI - Disturbance of cultured rat neuronal network activity depends on concentration and ratio of leucine and alpha-ketoisocaproate: implication for acute encephalopathy of maple syrup urine disease. AB - Increased concentrations of leucine and its respective ketoacid alpha ketoisocaproate (KIC) in plasma and cerebrospinal fluid are related to acute and reversible encephalopathy in patients with maple syrup urine disease. We studied electrophysiological properties of primary dissociated rat neurons at increased extracellular concentrations of leucine and KIC (1-10 mM). Spontaneous neuronal network activity was reversibly reduced or blocked by leucine as well as by KIC in a dose-dependent manner. Simultaneous incubation with both substances led to a minor inhibition compared to the effect of each substance alone. Neuronal resting potential, voltage dependent Na(+) (I(Na)) and K(+) (I(K)) currents, the GABA- and glycine-elicited membrane currents, and glutamate-induced intracellular Ca(2+) increase of single neurons, however, were unaffected by both substances. We conclude that acute neuronal network dysfunction in maple syrup urine disease is mainly based on an imbalance of the presynaptic glutamatergic/GABAergic neurotransmitter concentrations or their release. PMID- 12538794 TI - Effect of reduced maternal inspired oxygen on hepatic glucose metabolism in the rat fetus. AB - Perturbations in glucose metabolism in the fetus and in the neonate are a consistent finding in several different animal models of intrauterine growth retardation (IUGR) as well as in humans. Studies in rats who have undergone IUGR have shown decreased hepatic glycogen stores in the fetus and delayed induction of cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) at birth. Hepatic transcription factors CCAAT enhancer binding protein (C/EBP)alpha and C/EBPbeta and the increase in cyclic AMP at birth have been implicated in the initial appearance of PEPCK-C. We have examined the effect of IUGR induced by reduced maternal inspired oxygen (fractional inspired oxygen concentration 0.14) on a) the expression of genes for hepatic C/EBPalpha, C/EBPbeta, PEPCK-C and glycogen synthase; and b) transcription of the genes for C/EBPbeta and PEPCK-C by dibutyryl cyclic AMP in the fetus. Three days (d 18-21) of decrease in maternal inspired oxygen resulted in lower maternal arterial PO(2) and a lower birth weight of the pups (p < 0.01). Fetuses that underwent IUGR had significantly lower concentrations of plasma glucose, hepatic glycogen, and glycogen synthase mRNA and a higher hepatic lactate:pyruvate ratio. They also had lower levels of hepatic PEPCK-C mRNA at birth. The concentration of hepatic mRNA for C/EBPalpha and C/EBPbeta as well as the transcription factors themselves were not affected by the decreased maternal inspired oxygen. Fetal injection of dibutyryl cyclic AMP after 24 h of decreased maternal inspired oxygen (d 18-19) had no effect on the expression of C/EBPbeta. However, it resulted in an attenuated induction of PEPCK-C in the fetuses with IUGR. We speculate that a decrease in maternal inspired oxygen induced certain mediators, either in the mother or in the placenta, that caused lower fetal glucose concentration and affected the transcription of genes involved in fetal hepatic glucose metabolism. IUGR, as a result of decreased fractional inspired oxygen concentration may also be the consequence of pH-mediated changes in uterine blood flow. However, these remain to be examined in this experimental model. PMID- 12538795 TI - Fibrinolytic system in adolescents: response to venous occlusion stress tests. AB - Recent studies indicate that the incidence of thromboembolic events is increasing as a secondary complication in children with serious underlying diseases. The mechanism to eliminate these thrombi via the thrombolytic system in children is unknown. The baseline fibrinolytic system is age dependent, with significant variation between children and adults. Adult studies would suggest that the fibrinolytic response to venous occlusion has more clinical relevance than the baseline fibrinolytic system. The aim of this study was to determine whether the fibrinolytic response to venous occlusion stress testing in healthy adolescents differs from the response in healthy adults. Healthy adolescents (13-18 y) from a school population and normal adults were recruited. Pre- and postvenous occlusion blood samples were collected using standard techniques. Plasma tissue plasminogen activator, plasminogen activator inhibitor-1, plasminogen, alpha(2)-antiplasmin, alpha(2)-macroglubulin, D-dimers, euglobulin lysis time, and fibrinogen were measured on each sample. Adolescents had significantly decreased tissue plasminogen activator antigen levels and increased plasminogen activator inhibitor-1 activity levels after venous occlusion, resulting in significantly prolonged euglobulin lysis times. The results of our study confirm developmental differences in the fibrinolytic response to venous occlusion stress testing. The age-related differences in fibrinolytic response to venous occlusion of younger children and the significance of these differences on the pathophysiology of thromboembolic events in children require further studies. PMID- 12538797 TI - Child neurology in the 20th century. AB - Although considered a relatively new subspecialty, child neurology traces its origins to the Hippocratic descriptions of seizures and other neurologic conditions in children. Its true beginnings can be traced to the 1600s and 1700s with classical descriptions of chorea, hydrocephalus, spina bifida, and polio. It was, however, the remarkable clinical and scientific advances in neurology and pediatrics at the end of the 19th century that helped create its scientific foundation. Like other pediatric disciplines, child neurology evolved into a distinct clinical and scientific specialty early in the 20th century. Remarkable advances in the neurosciences, particularly in the fields of genetics, molecular biology, metabolism, immunology and nutrition, have greatly advanced our understanding of how the brain develops and responds to environmental influences. Advances in neuroimaging, electroencephalography, electromyography, muscle histology, biochemistry, and neuropharmacology have considerably improved our ability to evaluate and treat children with neurological disorders. These advances have allowed new and expanding approaches, unique to children, in the fields of epilepsy, neurodegenerative and neurometabolic disorders, nervous system infections, demyelinating diseases and tumors, neonatal neurological conditions, and neuromuscular diseases. They have also led to a better understanding of the neurobiologic basis of common problems such as global developmental delay, cerebral palsy, and autism. As remarkable as the advances have been in the past century, the accelerating pace of our understanding of the fundamental mechanisms responsible for brain development will lead to even greater achievements in the clinical care of children with neurological disorders in the 21st century PMID- 12538796 TI - Minimum protein intake for the preterm neonate determined by protein and amino acid kinetics. AB - Lower limits of protein needs in prematurely born neonates have not been adequately studied, yet providing protein in amounts maximizing accretion without excess is a goal in these infants' nutritional care. We hypothesized that with the use of amino acid oxidation methodology, it would be possible to define minimum protein requirement. Our objective was to investigate protein kinetics during short-term changes in protein intake by measurement of nitrogen balance and amino acid flux and oxidation using [(15)N]glycine, [(13)C]phenylalanine, and [(13)C]leucine tracers. Protein kinetics were examined in 21 preterm infants (gestational age: 29 +/- 3 wk; birth weight: 1091 +/- 324 g) at five protein intakes (1.0, 1.5, 2.0, 2.5, and 3.0 g x kg(-1) x d(-1)) with 1 d of adaptation to the test intakes. From nitrogen balance data, a protein need of 0.74 g x kg(-1 x -1) was estimated to achieve zero balance. For all three amino acids, flux and oxidation estimates were not different across protein intakes. Whole-body protein synthesis and breakdown estimates from [(15)N]ammonia data were 14.6 +/- 3.4 and 14.4 +/- 4.1 g x kg(-1) x d(-1), respectively. Glycine flux (680 +/- 168 micromol x kg(-1) x h(-1)) was greater than leucine flux (323 +/- 115 micromol x kg(-1) x h(-1)), which was greater than phenylalanine flux (84.3 +/- 35.2 micromol x kg( 1) x h(-1)). Leucine oxidation (36.7 +/- 15.6 micromol x kg(-1) x h(-1)) was also greater than phenylalanine oxidation (6.64 +/- 4.41 micromol x kg(-1) x h(-1)). Infants in our study were able to adapt to short-term changes in protein intake with little consequence to the overall whole-body protein economy, as measured by the three test amino acids. PMID- 12538798 TI - Experimental therapeutics in the Renaissance. AB - Detailed accounts of therapeutics at the time of the European Renaissance written by the participants have not survived in large numbers. One manuscript, dated 1562, was written by friars in a religious order in Italy dedicated to the care of the sick. Their remedies, methods of preparation, and uses were detailed by the friars and offer a glimpse into the beginnings of experimentation with drugs and rejection of tradition and authority in determining the effectiveness of a remedy. These developing concepts were combined in the manuscript with traditional treatments dating back through the Middle Ages to the medical methods of Greece and Rome. PMID- 12538799 TI - Proteolytic degradation of nitric oxide synthase: effect of inhibitors and role of hsp90-based chaperones. AB - Nitric oxide synthase (NOS) is a highly regulated enzyme that produces nitric oxide, a critical messenger in many physiological processes. In this perspective, we explore the role of proteolytic degradation of NOS, in particular the inducible and neuronal isoforms of NOS, as a mechanism of regulation of the enzyme. The ubiquitin-proteasome and calpain pathways are the major proteolytic systems identified to date that are responsible for this regulated degradation. The degradation of NOS is affected by diverse agents, including glucocorticoids, caveolin, neurotoxic compounds, and certain NOS inhibitors. Some irreversible inactivators of NOS enhance the proteolytic degradation of the enzyme, and this property may be of great importance in understanding the biological effects of these inhibitors, some of which are being developed for clinical use. Analogies with the regulated degradation of liver microsomal cytochromes P450, which are related to NOS, provide a framework for understanding these processes. Finally, a new perspective on the regulation of NOS by hsp90-based chaperones is presented that involves facilitated heme insertion into the enzyme. PMID- 12538800 TI - Pharmacokinetics and pharmacodynamics of entacapone and tolcapone after acute and repeated administration: a comparative study in the rat. AB - Two catechol-O-methyltransferase (COMT) inhibitors, entacapone and tolcapone, were compared in the rat to elucidate the actual differences between their pharmacokinetics and pharmacodynamics after single and repeated administration. Their inhibitory potencies were also compared in vitro. After intravenous administration (3 mg/kg), the elimination half-life (t(1/2 beta)) of entacapone (0.8 h) was clearly shorter than that of tolcapone (2.9 h). The striatum/serum ratio of tolcapone was 3-fold higher than that of entacapone. After a single oral dose (10 mg/kg), both entacapone and tolcapone produced an equal maximal degree of COMT inhibition in peripheral tissues, but tolcapone inhibited striatal COMT more effectively than did entacapone. After the 7-day treatment (10 mg/kg twice daily), COMT activity had recovered to a level of 67 to 101% of control within 8 h after the last dose of entacapone. In tolcapone-treated animals, there was still extensive COMT inhibition present in peripheral tissues, and the degree of inhibition was higher than that attained after a single dose. The pharmacokinetic pharmacodynamic modeling revealed that a plateau of COMT inhibition near the maximal attainable inhibition was reached already by plasma concentrations below 2000 ng/ml, both with entacapone and tolcapone. Entacapone and tolcapone inhibited equally rat liver COMT in vitro with K(i) values of 10.7 and 10.0 nM, respectively. In conclusion, tolcapone has a longer duration of action and a better brain penetration than entacapone. The results also suggest that peripheral COMT is inhibited continuously when tolcapone is dosed at 12-h intervals, but this was not seen with entacapone. PMID- 12538801 TI - Contributions of nitric oxide and prostanoids and their signaling pathways to the renal medullary vasodilator effect of U46619 (9-11-dideoxy-11 alpha,9a epoxymethano-prostaglandin F(2a)) in the rat. AB - We recently demonstrated that U46619 (9-11-dideoxy-11 alpha,9a-epoxymethano prostaglandin F(2a)) evoked a medullary vasodilation and a reduction in blood pressure despite a potent cortical vasoconstriction in the anesthetized rat. The present study tested the hypothesis that nitric oxide (NO) and prostanoids contribute to U46619-induced increase in medullary blood flow (MBF). U46619 at 1, 3, and 5 microg/kg increased MBF (above basal values) by 16 +/- 3, 45 +/- 10, and 58 +/- 8 perfusion units, respectively, and increased NO current in the medulla by 17 +/- 4, 34 +/- 7, and 60 +/- 12 pA, respectively. N(omega)-L-Nitro-arginine methyl ester (5 mg/kg), the inhibitor of NO production, attenuated the increase in MBF (75 +/- 8%, p < 0.05) as did indomethacin (10 mg/kg), the inhibitor of cyclooxygenase (38 +/- 5%, p < 0.05), suggesting the involvement of NO and dilator prostanoids. H-Arg-Lys-Arg-Ala-Arg-Lys-Glu-OH, a synthetic peptide and selective inhibitor of cGMP-dependent protein kinase, attenuated U46619-induced medullary perfusion (52 +/- 6%, p < 0.05), but H-89 ((N-[2-((p bromocinnamyl)aminoethyl)]-5-isoquinolinesulfonamide hydrochloride), a cell permeable, selective, and potent inhibitor of cAMP-dependent protein kinase A, was without effect. Glybenclamide, a K(ATP) channel blocker, also blunted the increase by U46619 in MBF (58 +/- 7%, p < 0.05). These data suggest that NO and prostanoids contribute to U46619-induced medullary perfusion and that the effects of these mediators are coupled to activation of protein kinase G and K(ATP) channels but not protein kinase A. PMID- 12538802 TI - Persistent effects of delayed treatment with nefiracetam on the water maze task in rats with sustained cerebral ischemia. AB - The present study was aimed at determining whether nefiracetam might have a persistent cognition-enhancing effect in animals with sustained cerebral ischemia. Sustained cerebral ischemia was induced by injecting 700 microspheres into the right internal carotid artery of rats [microsphere-embolized (ME) rats]. The ME and sham-operated rats were treated with 10 mg/kg/day nefiracetam p.o. from the first to the 9th day after the operation. The escape latency of the ME rat in the water maze test, when performed on days 7 to 9 after the operation, was lengthened. This effect was attenuated by the delayed treatment with nefiracetam. The nefiracetam-treated ME rat showed a shortened escape latency in the retention test on day 17 as well as in the contraposition test on day 18. These results indicate that a persistent improvement of the spatial memory function impaired by sustained cerebral ischemia was achieved even after cessation of treatment with nefiracetam. The functional damage to learning and memory was associated with decreases in the membranous adenylyl cyclase I and cytosolic protein kinase A (PKA) catalytic subunit and regulatory subunit proteins in the right hippocampus and cerebral cortex. The delayed treatment with nefiracetam appreciably prevented the decreases in these proteins. The present study suggests that nefiracetam may have an ability to cause persistent improvement of learning and memory function, possibly through protection against the ischemia-induced impairment to the adenylyl cyclase/cAMP/PKA signal transduction pathway. PMID- 12538803 TI - Influence of omeprazole on multidrug resistance protein 3 expression in human liver. AB - Multidrug resistance protein (MRP) 3 transports bile salts and conjugated xenobiotics from cells (hepatocytes and enterocytes) into the blood. Hepatic MRP3 expression is low under normal conditions but is markedly up-regulated during cholestasis. Since little is known about additional factors increasing human hepatic MRP3 expression, we investigated the variability of MRP3 expression in a large collection of human livers and factors contributing to variable MRP3 expression in liver and HepG2 cells. MRP3 was measured in 62 human livers from patients with and without omeprazole treatment and in HepG2 cells with and without omeprazole or beta-naphthoflavone treatment. Livers of patients treated with omeprazole showed 4.8-fold (P < 0.0001) higher MRP3 protein expression compared with the remainder of the population. Accordingly, MRP3 mRNA and protein were induced 2.4- and 1.8-fold, respectively (P < 0.01 and P < 0.05), in HepG2 cells treated with omeprazole. Finally, MRP3 was induced in HepG2 cells by beta naphthoflavone. In summary, treatment with omeprazole and beta-naphthoflavone is a determinant of variable human hepatic MRP3 expression. PMID- 12538804 TI - Kv1.1 channels of dorsal root ganglion neurons are inhibited by n-butyl-p aminobenzoate, a promising anesthetic for the treatment of chronic pain. AB - In this study, we investigated the effects of the local anesthetic n-butyl-p aminobenzoate (BAB) on the delayed rectifier potassium current of cultured dorsal root ganglion (DRG) neurons using the patch-clamp technique. The majority of the K(+) current of small DRG neurons rapidly activates and slowly inactivates at depolarized voltages. BAB inhibited the whole-cell K(+) current of these neurons with an IC(50) value of 228 microM. Dendrotoxin K (DTX(K)), a specific inhibitor of Kv1.1, reduced the DRG K(+) current at +20 mV by 34%, consistent with an important contribution of channels incorporating the Kv1.1 subunit to the delayed rectifier current. To further investigate the mechanism of BAB inhibition, we examined its effect on Kv1.1 channels heterologously expressed in mammalian tsA201 cells. BAB inhibits the Kv1.1 channels with an IC(50) value of 238 microM, similar to what was observed for the native DRG current. BAB accelerates the opening and closing of Kv1.1, but does not alter the midpoint of steady-state activation. BAB seems to inhibit Kv1.1 by stabilizing closed conformations of the channel. Coexpression with the Kv beta 1 subunit induces rapid inactivation and reduces the BAB sensitivity of Kv1.1. Comparison of the heterologously expressed Kv1.1 and native DRG currents indicates that the Kv beta 1 subunit does not modulate the gating of the DTX(K)-sensitive Kv1.1 channels of DRG neurons. Inhibition of the delayed rectifier current of these neurons may contribute to the long-duration anesthesia attained during the epidural administration of BAB. PMID- 12538805 TI - The relative contribution of monoamine oxidase and cytochrome p450 isozymes to the metabolic deamination of the trace amine tryptamine. AB - Tryptamine is a trace amine in mammalian central nervous system that interacts with the trace amine TA(2) receptor and is now thought to function as a neurotransmitter or neuromodulator. It had been reported that deamination of tryptamine to tryptophol was mediated by CYP2D6, a cytochrome P450 that is expressed in human brain, suggesting that tryptamine may be an endogenous substrate for this polymorphic enzyme. We were unable to confirm this report and have reinvestigated tryptamine metabolism in human liver microsomes (HLM) and in microsomes expressing recombinant human cytochrome P450 and monoamine oxidase (MAO) isozymes. Tryptamine was oxidized to indole-3-acetaldehyde by HLM and recombinant human MAO-A in the absence of NADPH, and indole-3-acetaldehyde was further reduced to tryptophol by aldehyde reductase in HLM in the presence of NADPH. Steady-state kinetic parameters were estimated for each reaction step by HLM and MAO-A. The CYP2D6 substrates bufuralol and debrisoquine showed strong inhibition of both tryptophol production from tryptamine in HLM and the formation of indole-3-acetaldehyde from tryptamine catalyzed by recombinant MAO-A. Anti CYP2D6 monoclonal antibody did not inhibit these reactions. Pargyline, a nonselective MAO inhibitor, did not show cross inhibition to debrisoquine 4 hydroxylation and dextromethorphan O-demethylation by HLM and recombinant CYP2D6 enzyme. This is the first unequivocal report of the selective conversion of tryptamine to tryptophol by MAO-A. CYP2D6 does not contribute to this reaction. PMID- 12538806 TI - The heritability of antinociception: common pharmacogenetic mediation of five neurochemically distinct analgesics. AB - The heritability of nociception and antinociception has been well established in the mouse. The pharmacogenetics of morphine analgesia are fairly well characterized, but far less is known about other analgesics. The purpose of this work was to begin the systematic genetic study of non-mu-opioid analgesics. We tested mice of 12 inbred mouse strains for baseline nociceptive sensitivity (49 degrees C tail-withdrawal assay) and subsequent antinociceptive sensitivity to systemic administration of (trans)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl) cyclohexyl]benzeneacetamide methane-sulfonate hydrate (U50,488; 10-150 mg/kg), a kappa-opioid receptor agonist; (R)-(+)-[2,3-dihydro-5-methyl-3-(4 morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-naphthalenylmethanone (WIN55,212-2; 0.5-480 mg/kg), a synthetic cannabinoid receptor agonist; epibatidine (7.5-150 microg/kg), a nicotinic receptor agonist; clonidine (0.1-5 mg/kg), an alpha(2)-adrenergic receptor agonist; and, for purposes of comparison, the prototypic mu-opioid receptor agonist, morphine (5-200 mg/kg). Robust interstrain variability was observed in nociceptive sensitivity and in the antinociceptive effects of each of the drugs, with extreme-responding strains exhibiting antinociceptive potencies differing up to 37-fold. Unexpectedly, we observed moderate-to-high genetic correlations of strain sensitivities to the five drugs (r = 0.39-0.77). We also found moderate-to-high correlations between baseline nociceptive sensitivity and subsequent analgesic response to each drug (r = 0.33-0.68). The generalizability of these findings was established in follow up experiments investigating morphine and clonidine inhibition of formalin test nociception. Despite the fact that each drug activates a unique receptor, our results suggest that the potency of each drug is affected by a common set of genes. However, the genes in question may affect antinociception indirectly, via a primary action on baseline nociceptive sensitivity. PMID- 12538807 TI - Interaction of cysteine conjugates with human and rabbit organic anion transporter 1. AB - Organic anion (OA) transport mediates accumulation of the zwitterionic nephrotoxic cysteine S-conjugates S-dichlorovinylcysteine (DCVC) and S chlorotrifluoroethylcysteine (CTFC) in the rabbit renal proximal tubule (RPT). Although these cysteine conjugates are nephrotoxic to the human RPT, neither the role of OA transport nor the specific OA transport pathway(s) involved in cysteine conjugate accumulation are known. Since the OAT1 transporter has the characteristics of para-aminokippurate (PAH) transport that closely correlate to the native RPT, we examined the interaction of DCVC, CTFC, and the nontoxic benzothiazolylcysteine (BTC) with PAH transport mediated by human OAT1 and rabbit Oat1 expressed in Chinese hamster ovary and COS7 heterologous expression systems, respectively. Although the K(m) values for PAH uptake by hOAT1 and rbOat1 (8.9 +/ 3.6 and 20.7 +/- 8 microM, respectively) were 5- to 10-fold less than the K(m) for peritubular PAH transport into rabbit RPT, the IC(50) values for DCVC, CTFC, and BTC inhibition of PAH uptake mediated by either hOAT1 or rbOat1 were similar between these two transporters and to the IC(50) values for these conjugates measured in rabbit RPT. The IC(50) for inhibition of hOAT1- and rbOat1-mediated PAH uptake by the hydrophobic conjugate BTC was more than 5-fold lower than the IC(50) values seen with DCVC and CTFC, suggesting that hydrophobicity increases the affinity of OAT1 for cysteine conjugates. Finally, preloading cells transfected with hOAT1 with BTC significantly trans-stimulated the uptake of PAH, consistent with the conclusion that BTC and, hence, other cysteine S-conjugates are substrates for hOAT1. PMID- 12538808 TI - Antithrombotic properties of SSR182289A, a new, orally active thrombin inhibitor. AB - N-[3-[[[(1S)-4-(5-Amino-2-pyridinyl)-1-[[4-difluoromethylene)-1 piperidinyl]carbonyl]butyl]amino]sulfonyl][1,1'-biphenyl]-2-yl]acetamide hydrochloride (SSR182289A) is a novel, potent, and selective thrombin inhibitor. We have examined the antithrombotic properties of SSR182289A administered by i.v. and p.o. routes in several different animal thrombosis models in comparison with reference antithrombotic agents. Oral administration of SSR182289A produced dose related antithrombotic effects in the following models; rat venous thrombosis (ED(50) 0.9 mg/kg p.o.), rat silk thread arterio-venous (AV) shunt (ED(50) 3.8 mg/kg p.o.), rat thromboplastin-induced AV shunt (ED(50) 3.1 mg/kg p.o.), rat carotid artery thrombosis (ED(200) 5.9 mg/kg p.o.), and rabbit venous thrombosis (ED(50) 7.5 mg/kg p.o.). Administered as an i.v. bolus, SSR182289A showed antithrombotic activity in the above models with ED(50)/ED(200) values in the range of 0.2 to 1.9 mg/kg i.v. SSR182289A increased rat tail transection bleeding time at doses > or =10 mg/kg p.o. In the rat thromboplastin-induced AV shunt model, SSR182289A 10 mg/kg p.o. produced marked antithrombotic effects at 30, 60, 120, and 240 min after administration. Hence, SSR182289A demonstrates potent oral antithrombotic properties in animal venous, AV-shunt, and arterial thrombosis models. PMID- 12538809 TI - Chronic nicotine treatment leads to sustained stimulation of tyrosine hydroxylase gene transcription rate in rat adrenal medulla. AB - Nicotine is a powerful stimulant of the sympathoadrenal system, causing the release of peripheral catecholamines and activation of catecholamine biosynthesis. In previous reports, we have studied the mechanisms by which short term nicotine treatment regulates tyrosine hydroxylase (TH) in adrenal medulla. In this report, we study the effects of chronic nicotine treatment on adrenal TH gene expression. Rats were injected with either saline or nicotine twice per day for up to 14 days. Chronic nicotine treatment elicited long-lasting, dose dependent increases in the levels of adrenal TH mRNA, TH protein, and TH activity. In contrast, a single injection of nicotine elicited only a small increase in adrenal TH mRNA levels, which was transient and did not result in the induction of TH enzyme. Chronic nicotine administration also elicited a sustained increase in adrenal TH gene transcription rate, which persisted for up to 7 days after the final nicotine injection. This sustained transcriptional response correlated with a modest sustained increase in adrenal TH AP1 binding, but not in the levels of Fra-2 or other fos or jun proteins. These results demonstrate that repeated nicotine injections administered chronically over 1 to 2 weeks lead to sustained stimulation of the TH gene and consequent induction of TH gene expression in rat adrenal medulla. These studies support the hypothesis that chronic nicotine administration produces long-lasting cellular changes in adrenal medulla that lead to sustained transcriptional responses. PMID- 12538810 TI - Inhibition of prostaglandin H2 synthases by salicylate is dependent on the oxidative state of the enzymes. AB - At antipyretic and analgesic doses, salicylate has no antiplatelet or anti inflammatory effects, unlike typical inhibitors of the prostaglandin H synthases (PGHSs). We demonstrated that salicylate inhibits PGHS-1 and -2 with a potency inversely related to ambient hydroperoxide concentrations. Salicylate inhibition of PGHS-1 was prevented by 12-hydroperoxyeicosatetraenoic acid (12-HPETE). Increasing the production of prostaglandin G2 (the peroxide product of PGHS cyclooxygenase activity) by elevating the concentration of either enzyme or substrate reversed inhibition. Using analogs of benzoic acid differing only at the hydroxyl position, we revealed the importance of this moiety to salicylate's inhibitory mechanism. Unlike typical phenolic inhibitors, e.g., acetaminophen, salicylate was ineffective as a reducing cosubstrate for PGHS-peroxidase activity, implicating the cyclooxygenase site as its putative target. PGHS cyclooxygenase activity depends upon the oxidation of an active site tyrosine by electron transfer to the oxidized ferriprotoporphyrin of the peroxidase. The PGHS 1 apoenzyme reconstituted with manganese protoporphyrin instead of iron protoporphyrin has very little peroxidase activity. 12-HPETE does not prevent the inhibition of Mn-PGHS-1 by salicylate, indicating that reversal of salicylate inhibition by hydroperoxides depends upon electron transfer between the cyclooxygenase and peroxidase active sites. These results are consistent with an inhibitory action of salicylate at the PGHS-cyclooxygenase site that is dependent on the PGHS-peroxidase activity. PMID- 12538811 TI - Assessing safety and efficacy of directed P-glycoprotein inhibition to improve the pharmacokinetic properties of saquinavir coadministered with ritonavir. AB - Using a mouse model, we tested the effects of in vivo P-glycoprotein inhibition to enhance the oral uptake and penetration into pharmacological sanctuary sites of the human immunodeficiency virus protease inhibitor (HPI) saquinavir. The HPI ritonavir is frequently coadministered with saquinavir to improve saquinavir plasma levels since it strongly reduces the cytochrome P450 3A4-mediated metabolism of saquinavir. Previously, we demonstrated that ritonavir is not an efficient P-glycoprotein inhibitor in vivo, evidenced by the limited oral uptake of saquinavir and its penetration into brain and fetus. Increasing drug concentrations in these sites using more effective P-gp inhibitors might improve therapy but could also lead to toxicity. We orally coadministered ritonavir and saquinavir to mice, with or without the potent P-glycoprotein inhibitor N-(4-[2 (1,2,3,4-tetrahydro-6,7-dimethoxy-2-isoquinolinyl)ethyl]-phenyl)-9,10-dihydro-5 methoxy-9-oxo-4-acridine carboxamide (GF120918). Upon GF120918 coadministration, two of seven P-glycoprotein-deficient animals died. Using a decreased ritonavir dose, GF120918 coadministration led to a 4.4-fold increase in the saquinavir plasma area under the curve in wild-type mice, whereas no such effect was observed in P-glycoprotein-deficient mice. Despite the decreased ritonavir dose, all mice did suffer from impaired gastric emptying. Including GF120918 in a multiple (twice daily) dosing regimen, we found continued accumulation of saquinavir in brain over several days, resulting in 10-fold higher levels compared with vehicle-treated mice. Transient ritonavir-related neurotoxicity, however, was observed after the fourth and final drug dosing. Clinical attempts to efficiently inhibit P-glycoprotein function for improved HPI disposition may therefore be feasible, but they should be performed without ritonavir and monitored carefully for unexpected toxicities. PMID- 12538812 TI - Changes in urination/defecation, auditory startle response, and startle-induced ultrasonic vocalizations in rats undergoing morphine withdrawal: similarities and differences between acute and chronic dependence. AB - In drug-free subjects, a single dose of morphine followed by an opioid antagonist a few hours later results in signs of a withdrawal syndrome, suggesting a state of physical dependence. Increased urination/defecation, altered startle, and ultrasonic vocalizations (USV) are some signs of the withdrawal syndrome in rats chronically dependent on morphine. We investigated whether naltrexone stimulates urination/defecation and alters startle and USV in male rats that were pretreated with only a single dose of morphine and compared these indices to the ones of chronic dependence. Separate groups of rats were pretreated with either a single dose (10 mg/kg) or with a continuous s.c. infusion of morphine via an osmotic pump. Naltrexone (0.01-1.0 mg/kg) was administered 2 to 6 h after the single dose of morphine and on days 7 to 11 of the infusion. Immediately after the naltrexone injection subjects were placed in sound-attenuating boxes to record startle and USV and to collect urine/feces. Subjects chronically exposed to morphine also were tested during spontaneous withdrawal 3 to 24 h after pump removal. Naltrexone increased urination/defecation in subjects pretreated with morphine either chronically or acutely; it increased startle and USV in acutely dependent rats but decreased them in chronically dependent rats. In the latter group, changes in the four variables during spontaneous withdrawal were qualitatively similar to those during precipitated withdrawal but smaller in magnitude. Differences in withdrawal signs between acute and chronic dependence suggest that the neural substrates that mediate those particular components of the withdrawal syndrome are affected differently in the two states of dependence. PMID- 12538813 TI - Inhibition of transporter-mediated hepatic uptake as a mechanism for drug-drug interaction between cerivastatin and cyclosporin A. AB - The mechanism involved in the clinically relevant drug-drug interaction (DDI) between cerivastatin (CER) and cyclosporin A (CsA) has not yet been clarified. In the present study, we examined the possible roles of transporter-mediated hepatic uptake in this DDI. The uptake of [(14)C]CER into human hepatocytes prepared from three different donors was examined. Kinetic analyses revealed K(m) values for the uptake of [(14)C]CER within the range of 3 to 18 microM, suggesting that more than 70% of the total uptake at therapeutic CER concentrations was accounted for by a saturable process, i.e., transporter-mediated uptake. This uptake was inhibited by CsA with K(i) values of 0.3 to 0.7 microM. The uptake of [(14)C]CER was also examined in human organic anion transporting polypeptide-2 (OATP2) expressing Madin-Darby canine kidney cells (MDCKII). Saturable OATP2-mediated uptake of [(14)C]CER was observed and was also inhibited by CsA, with a K(i) value of 0.2 microM. These results suggest that the DDI between CER and CsA involves the inhibition of transporter-mediated uptake of CER and, at least in part, its OATP2-mediated uptake. The effect of CsA on the in vitro metabolism of [(14)C]CER was also examined. The metabolism of [(14)C]CER was inhibited by CsA with an IC(50) value of more than 30 microM. From these results, we conclude that the DDI between CER and CsA is mainly due to the inhibition of transporter (at least partly OATP2)-mediated uptake in the liver. PMID- 12538814 TI - Neuroprotective effects of L-arginine administration after cortical impact injury in rats: dose response and time window. AB - Administration of L-arginine has been shown to increase cerebral blood flow and reduce neurological damage after experimental traumatic brain injury. The purpose of this study was to examine the optimal dose and time window for these neuroprotective effects. In a dose response experiment, doses of L-arginine ranging from 37.5 to 600 mg/kg were administered 5 min after a 5-m/s, 3-mm, controlled cortical impact in rats. The amount of brain injury found at 2 weeks after injury, both at the contusion site and in the ipsilateral hippocampus, were inversely related to the dose of L-arginine administered. Both 300- and 600-mg/kg doses of L-arginine significantly reduced contusion volume. The 300-mg/kg dose significantly increased the neuron density in the CA1 region of the hippocampus. Physiological effects of L-arginine were also dose-related. The greatest reduction in intracranial pressure occurred with the 300-mg/kg dose of L arginine. Doses up to 300 mg/kg were well tolerated, but the 600-mg/kg dose resulted in transient hypotension. In another experiment, 300 mg/kg L-arginine was administered at times varying from 5 min to 48 h after injury. Contusion volume was significantly reduced when the L-arginine was given at 5 min and 1 h after injury. The protective effect was less when the same dose was given at the later times, but there was no evidence of an adverse effect even when the L arginine was administered 48 h after injury. PMID- 12538815 TI - Histamine inhibits lipopolysaccharide-induced tumor necrosis factor-alpha production in an intercellular adhesion molecule-1- and B7.1-dependent manner. AB - Lipopolysaccharide (LPS) is recognized as a key molecule in the pathogenesis of Gram negative sepsis and septic shock. In the present study, we demonstrate that LPS (1-1000 pg/ml) concentration dependently up-regulated the expression of intercellular adhesion molecule (ICAM)-1, B7.1, and B7.2 on human monocytes using fluorescence-activated cell sorting analysis, and that tumor necrosis factor (TNF)-alpha production induced by LPS in peripheral blood mononuclear cells (PBMCs) was inhibited by the addition of antibodies against these adhesion molecules, suggesting the dependence of TNF-alpha production on cell-cell interaction through these adhesion molecules. Moreover, we found that histamine (10(-7)-10(-4) M) concentration dependently inhibited the expression of ICAM-1 and B7.1, but not B7.2 on monocytes induced by LPS. Histamine also inhibited the responses of TNF-alpha production induced by LPS. The modulatory effects of histamine on ICAM-1 and B7.1 expression and TNF-alpha production were all concentration dependently antagonized by famotidine but not by d-chlorpheniramine and thioperamide, and were mimicked by selective H2-receptor agonists but not by H1-, H3-, and H4-receptor agonists, indicating the involvement of H2-receptors in the histamine action. Dibutyryl cAMP down-regulated ICAM-1 and B7.1 expression on monocytes stimulated by LPS, suggesting the mediation by the cyclic adenosine monophosphate-protein kinase A pathway of H2-receptor activation. These results as a whole indicated that histamine via H2-receptor inhibited the LPS-induced TNF alpha production through the regulation of ICAM-1 and B7.1 expression, leading to the reduction of innate immune response stimulated by LPS. PMID- 12538816 TI - Effect of beta 2-adrenergic receptor stimulation on interleukin-18-induced intercellular adhesion molecule-1 expression and cytokine production. AB - beta-Adrenergic receptor (AR) agonists have been demonstrated to modulate the production of inflammatory mediators. Recent studies implied that beta 2-AR agonists might be useful for chronic inflammatory diseases caused by interleukin (IL)-18. In the present study, we found that norepinephrine, epinephrine, or isoproterenol down-regulated IL-18 (100 ng/ml)-induced intercellular adhesion molecule (ICAM)-1 expression on monocytes in a dose-dependent manner (10(-8)-10( 4) M), but did not effect B7.1 and B7.2 expression after 24-h incubation. The modulatory effect of these catecholamines on ICAM-1 expression was antagonized by beta 2-AR antagonist, but not by alpha 1-, alpha 2-, or beta 1-AR antagonist. beta 2-AR-selective agonists salbutanol and terbutaline down-regulated IL-18 induced ICAM-1 expression on monocytes, but alpha 1-, alpha 2-, or beta1-AR agonist had no effect. In the same manner, salbutanol and terbutaline as well as norepinephrine, epinephrine, and isoproterenol regulated the IL-18-induced cytokine production, including IL-12, tumor necrosis factor-alpha or interferon gamma through the stimulation of beta 2-AR. Together with the previous finding that ICAM-1/lymphocyte function-associated antigen-1 interaction plays a crucial role in the IL-18-initiated cytokine network, the present study strongly suggested that the stimulation of beta 2-AR inhibited the IL-18-activated cytokine cascade through the inhibitory effect on ICAM-1 expression, contributing to finding a new method for clinical treatment. PMID- 12538817 TI - Structural requirements for functional interaction of glutathione tripeptide analogs with the human multidrug resistance protein 1 (MRP1). AB - The human multidrug resistance protein 1 (MRP1) is a primary active transporter of reduced (GSH) and oxidized glutathione, as well as GSH-, glucuronate-, and sulfate-conjugated organic anions. In addition, the transport of certain MRP1 substrates is stimulated by the presence of GSH. To evaluate the structural features of GSH required for interaction with the protein, we investigated the ability of a series of GSH analogs to enhance GSH stimulatable transport of [(3)H]estrone 3-sulfate (E(1)SO(4)). We found that substitution of the gamma-Glu residue with Gly, beta-Asp, and alpha-Glu resulted in complete loss of transport stimulation. In contrast, substitution of Gly with Glu or beta-Ala resulted in only a partial loss of stimulatory activity. E(1)SO(4) transport activity surpassed GSH-stimulated levels in the presence of tripeptides in which Cys was substituted with the hydrophobic amino acids Leu, Phe, and homo-Phe. Moreover, polar substitutions of Cys did not enhance transport to the same extent as nonpolar substitutions of comparable size. gamma-Glu-Leu-Gly was 1.6-fold more effective than GSH in stimulating E(1)SO(4) uptake, and kinetic analysis indicated this was due to an increased V(max). In addition, this tripeptide was shown to be a competitive inhibitor of apigenin-stimulated GSH transport (K(i) value of 14 microM), confirming that it either interacts with the same site on MRP1 as GSH or that the binding of the two tripeptides is mutually exclusive. These data provide insight into the architecture of the GSH binding domain of MRP1. PMID- 12538818 TI - Early treatment with hepatocyte growth factor improves cardiac function in experimental heart failure induced by myocardial infarction. AB - Plasma levels of hepatocyte growth factor (HGF) are increased within hours of cardiac ischemia/reperfusion in rats, and HGF has been shown to be cardioprotective toward acute ischemic injury. Myocardial levels of HGF mRNA and protein are increased for several days after myocardial infarction (MI), however, indicating a possible additional protective effect of HGF toward the progression of MI to heart failure. The purpose of this study was to determine whether HGF administration during the time course of endogenous cardiac HGF induction would lead to long-term improvement in cardiac function in rats with MI. MI was induced by 2-h occlusion of the left coronary artery, followed by reperfusion. HGF was given by intravenous infusion at 0.45 mg/kg/day for 6 days beginning on the day after surgery. Cardiac function and hemodynamic parameters were measured by using indwelling catheters and perivascular flow probes in conscious animals 8 weeks post-MI. Myocardial infarcts were approximately 30% of the left ventricle, and there was no difference in infarct size between the vehicle-treated and HGF treated groups. Compared with untreated sham-operated rats, vehicle-treated MI animals had significantly lower cardiac index and stroke volume index and higher systemic vascular resistance, indicating heart failure developed. Treatment with HGF caused a significant increase in cardiac index and stroke volume index and a reduction in systemic vascular resistance in rats with MI, restoring these parameters close to those observed in sham-operated control animals. These results provide direct evidence that HGF may be of benefit to cardiovascular function in ischemic cardiomyopathy. PMID- 12538819 TI - The effect of O6-alkylguanine-DNA alkyltransferase and mismatch repair activities on the sensitivity of human melanoma cells to temozolomide, 1,3-bis(2 chloroethyl)1-nitrosourea, and cisplatin. AB - The prognosis of advanced melanoma is generally poor, because this tumor commonly exhibits intrinsic or acquired resistance to chemotherapy. In an attempt to identify the underlying causes of this resistance, we studied the roles played by the DNA repair enzyme O(6)-alkylguanine-DNA alkyltransferase (OGAT) and the mismatch repair (MMR) system in the sensitivity of melanoma cells to temozolomide (TMZ), 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), or cis diamminedichloroplatinum(II) (CDDP). To this end, OGAT levels and MMR efficiency of extracts of nine melanoma cell lines and selected clones derived from four of these lines were determined and correlated with the sensitivity of the respective cells to these drugs. The effectiveness of O(6)-benzylguanine (BG), a specific OGAT inhibitor, in potentiating TMZ- or BCNU-mediated cytotoxicity was also evaluated. Our results demonstrate that MMR efficiency and OGAT levels strongly affect melanoma cell sensitivity to TMZ. In MMR-proficient cells, a direct correlation between OGAT levels and TMZ IC(50) values was found. When OGAT activity was inhibited with BG, the sensitivity of these cells to TMZ increased and was then dictated largely by their MMR efficiency. MMR-deficient cells were highly resistant to the drug irrespective of their OGAT levels. Although OGAT activity and MMR status seemed to be the major determinants of melanoma sensitivity to TMZ, this was not the case for BCNU and CDDP; resistance to the latter drugs clearly involves processes other than the two DNA repair pathways analyzed in this study. PMID- 12538820 TI - Optimizing the dosing schedule of TNP-470 [O-(chloroacetyl-carbamoyl) fumagillol] enhances its antitumor and antiangiogenic efficacies. AB - Many drugs vary in potency and/or toxicity according to the time of day when they are administered. In this study, we investigated whether antitumor efficacy of angiogenesis inhibitor, TNP-470 [O-(chloroacetyl-carbamoyl) fumagillol], could be improved by optimizing the dosing schedule. Tumor-bearing mice were housed under standardized light/dark cycle conditions (lights on at 7:00 AM, off at 7:00 PM) with food and water ad libitum. The antitumor effect of TNP-470 (30 mg/kg s.c.) was more potent in mice injected with the drug at the early light phase than it was when administered at the early dark phase. The diurnal change in the antitumor effect of TNP-470 was parallel to that in its antiangiogenic activity. The variation in the effects of TNP-470 was closely related to the diurnal variations in its inhibitory action on methionine aminopeptidase activity in tumor masses. There was a significant dosing time-dependent change in the concentration of TNP-470 in plasma. The higher concentration of TNP-470 in plasma was observed when its antitumor and antiangiogenic activities were increased. These results suggest that therapeutic efficacy of TNP-470 can be enhanced by choosing the most appropriate time of day to administer the drug. PMID- 12538821 TI - M1 receptor-mediated nitric oxide-dependent relaxation unmasked in stomach fundus from M3 receptor knockout mice. AB - Muscarinic receptors can mediate both contractile and relaxant responses in smooth muscle. The stomach fundus from wild-type mice possesses a neuronal M(1) receptor that mediates relaxation to carbamylcholine and (4-hydroxy-2-butynyl)-1 trimethylammonium-3-chlorocarbanilate chloride (McN-A-343) but is masked by M(3) receptor-mediated contraction to both agonists. When the M(3) receptor was deleted, cholinergic-induced relaxation was unmasked. M(1) receptor antagonism with pirenzepine, nitric oxide (NO) synthase inhibition with N(omega)-nitro-L arginine methyl ester hydrochloride, and inhibition of neuronal activation with tetrodotoxin abolished relaxation to McN-A-343 in tissues from M(3) receptor knockout mice, supporting the neuronal localization of an M(1) receptor that activated NO release to effect relaxation. However, the cyclooxygenase inhibitor indomethacin did not affect contraction or relaxation to carbamylcholine in stomach fundus from wild-type or M(3) receptor knockout mice, indicating that cyclooxygenase products played no role in these responses. The neuronal M(1) receptor modulated relaxation induced by carbamylcholine and McN-A-343 but not relaxation induced by electric field stimulation of the stomach fundus. These data support the presence of M(1) receptor-mediated relaxation in the stomach and suggest that when the M(3) receptor is eliminated or blocked, M(1) receptor mediated gastric relaxation may be enhanced, possibly leading to alterations in gastric emptying and subsequent effects on body weight. PMID- 12538822 TI - (2S,3R) beta-methyl-2',6'-dimethyltyrosine-L-tetrahydroisoquinoline-3-carboxylic acid [(2S,3R)TMT-L-Tic-OH] is a potent, selective delta-opioid receptor antagonist in mouse brain. AB - The constrained opioid peptide (2S,3R)beta-methyl-2',6'-dimethyltyrosine-L tetrahydroisoquinoline-3-carboxylic acid [(2S,3R)TMT-L-Tic-OH] exhibits high affinity and selectivity for the delta-opioid receptors (). In the present study, we examined the pharmacological properties of (2S,3R)TMT-L-Tic-OH in mouse brain. A 5'-O-(3-[(35)S]thiotriphosphate) ([(35)S]GTP gamma S) binding assay was used to determine the effect of (2S,3R)TMT-L-Tic-OH on G protein activity in vitro, in mouse brain membranes. delta- (SNC80; (+)-4-[(alpha R)-alpha-((2S,5R)-4-allyl-2,5 dimethyl-1-piperazinyl)-3-methoxy-benzyl]-N,N-diethyl-benzamide) or mu- (DAMGO; [D-Ala(2), Me-Phe(4),Gly(ol)(5)]enkephalin) selective opioid full agonists stimulated [(35)S]GTP gamma S binding in mouse brain membranes 150 +/- 4.5% and 152 +/- 5.7% over the basal level, respectively. (2S,3R)TMT-L-Tic-OH did not influence basal [(35)S]GTP gamma S binding in mouse brain membranes but dose dependently shifted the dose-response curve of SNC80 to the right, with a K(e) value of 3.6 +/- 0.7 nM. In contrast, (2S,3R)TMT-L-Tic-OH had no effect on the dose-response curve of the mu-selective opioid agonist, DAMGO. Warm water (55 degrees C) tail-flick and radiant heat paw-withdrawal tests were used to determine the in vivo nociceptive properties of (2S,3R)TMT-L-Tic-OH in the mouse. Intracerebroventricular injection of (2S,3R)TMT-L-Tic-OH had no significant effect on withdrawal latencies in either nociceptive tests. (2S,3R)TMT-L-Tic-OH (30 nmol/mouse) attenuated deltorphin II- but not DAMGO-mediated antinociception (40 +/- 13 and 100% of maximal possible effect, respectively) when administered intracerebroventricularly 10 min before the agonist. Taken together these results suggest that (2S,3R)TMT-L-Tic-OH is a potent highly selective neutral delta opioid antagonist in mouse brain. PMID- 12538823 TI - Role of phospholipase C-beta in the modulation of epithelial tight junction permeability. AB - The results presented in this study establish an association between phospholipase C-beta (PLC-beta) and tight junction permeability across Madin Darby canine kidney (MDCK) cell monolayers, an in vitro model for epithelial tissue. These results further show that PLC-beta modulates tight junction permeability by affecting actin filament organization. Hexadecylphosphocholine (HPC) inhibited PLC-beta and increased tight junction permeability in MDCK cells. Interestingly, the analogs of HPC, a series of alkylphosphocholines containing various lengths of linear alkyl chains, inhibited PLC-beta and increased tight junction permeability with a wide range of potency. The potency of alkylphosphocholines as enhancers of tight junction permeability significantly correlated (p < 0.05) with their potency as PLC-beta inhibitors. U73122, a steroid derivative that is structurally unrelated to alkylphosphocholines, inhibited PLC-beta and increased tight junction permeability with potencies that fit into the correlation observed for the alkylphosphocholine series. U73122 and HPC induced disorganization of actin filaments in MDCK cell monolayers. The potencies to cause disorganization of actin filaments were consistent with the potencies of these agents as inhibitors of PLC-beta and enhancers of tight junction permeability. Furthermore, ATP, an activator of PLC-beta, attenuated U73122-induced increase in tight junction permeability as well as disorganization of actin filaments. These results provide strong evidence that PLC-beta inhibition leads to increased tight junction permeability across MDCK cell monolayers through disorganization of actin filaments. PMID- 12538824 TI - p53-mediated regulation of expression of a rabbit liver carboxylesterase confers sensitivity to 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin (CPT-11). AB - We have exploited the ability of wild-type (wt) p53 to repress gene expression and produce tumor-selective cytotoxicity using viral-directed enzyme prodrug therapy. Vectors containing either the cytomegalovirus or Rous sarcoma virus promoter regulating transcription of a rabbit liver carboxylesterase (CE) have been constructed. Upon transfection of these plasmids into cells expressing either wt or mutant p53, differential expression of the CE has been observed, resulting in sensitization of the cells expressing the latter protein to the anticancer prodrug irinotecan, 7-ethyl-10-[4-(1-piperidino)-1-piperidino] carb- onyloxycamptothecin (CPT-11). Transduction of isogenic cell lines with adenovirus containing CE under control of the Rous sarcoma virus promoter confirmed the decreased sensitization of cells expressing wtp53 to CPT-11. These studies indicate that the inactivation of wtp53 by mutant p53 in human tumor cells may be sufficient enough to generate a therapeutic window for enhanced cytotoxicity with CPT-11. PMID- 12538825 TI - Papaverine blocks hKv1.5 channel current and human atrial ultrarapid delayed rectifier K+ currents. AB - Papaverine, 1-[(3,4-dimethoxyphenyl)methyl]-6,-7-dimethoxyisoquinoline, has been used as a vasodilator agent and a therapeutic agent for cerebral vasospasm, renal colic, and penile impotence. We examined the effects of papaverine on a rapidly activating delayed rectifier K(+) channel (hKv1.5) cloned from human heart and stably expressed in Ltk(-) cells as well as a corresponding K(+) current (the ultrarapid delayed rectifier, I(Kur)) in human atrial myocytes. Using the whole cell configuration of the patch-clamp technique, we found that papaverine inhibited hKv1.5 current in a time- and voltage-dependent manner with an IC(50) value of 43.4 microM at +60 mV. Papaverine accelerated the kinetics of the channel inactivation, suggesting the blockade of open channels. Papaverine (100 microM) also blocked I(Kur) in human atrial myocytes. These results indicate that papaverine blocks hKv1.5 channels and native hKv1.5 channels in a concentration-, voltage-, state-, and time-dependent manner. This interaction suggests that papaverine could alter cardiac excitability in vivo. PMID- 12538826 TI - Endocannabinoids induce ileitis in rats via the capsaicin receptor (VR1). AB - Intraluminal administration of the endocannabinoids N-arachidonoyl-ethanolamine (anandamide) and 2-arachidonoylglycerol (2-AG) causes inflammation similar to that caused by Clostridium difficile toxin A in the rat ileum. The effects of anandamide and 2-AG were significantly inhibited by pretreatment with the specific capsaicin receptor (vanilloid receptor subtype 1; VR1) antagonist capsazepine. Pretreatment with the CB1 and CB2 cannabinoid receptor antagonists N piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-3-pyrazole carboxamide (SR141716) and N-[1S)-endo-1,3,3-trimethylbicyclo[2.2.1]heptan-2-yl] 5-(4-chloro-3-methylphenyl)-1-(4-methylbenzyl)-pyrazole-3-carboxamide (SR144528) did not affect the responses to anandamide. It has previously been shown that intraluminal toxin A stimulates substance P (SP) release from primary sensory neurons and that pretreatment with SP receptor [neurokinin (NK)-1 receptor] antagonists inhibits the inflammatory effects of toxin A. Anandamide stimulated SP release and this was blocked by capsazepine pretreatment. Also, pretreatment with the specific NK-1 receptor antagonist (2S,3S)-3-([3,5 bis[trifluoromethyl)phenyl]methoxy)-2-phenylpiperidine (L-733,060) significantly inhibited the inflammatory effects of both toxin A and anandamide. Toxin A increased tissue concentrations of anandamide and 2-AG in the ileum, and these effects were enhanced after pretreatment with inhibitors of fatty acid amide hydrolase, a major endocannabinoid-degrading enzyme. The toxin A-stimulated release of anandamide but not 2-AG was selective over their congeners. These results demonstrate that the endocannabinoids anandamide and 2-AG stimulate intestinal primary sensory neurons via the capsaicin VR1 receptor to release SP, resulting in enteritis, and that endocannabinoids may mediate the inflammatory effects of toxin A. PMID- 12538827 TI - Inhibition of Rho-kinase reduces renal Na-H exchanger activity and causes natriuresis in rat. AB - Rho-kinase regulates the actin cytoskeleton and therefore modulates transport. The role of Rho-kinase in Na-H exchanger (NHE) activity of rat proximal convoluted tubules (PCTs) was investigated using (R)-(+)-trans-N-(4-pyridyl)-4-(1 aminoethyl)-cyclohexanecarboxamide (Y-27632), a specific inhibitor of Rho-kinase. In spontaneously hypertensive rats (SHR) and Wistar Kyoto (WKY) rats, apical and basolateral NHE activities were determined by measuring cell pH recovery following luminal NH4+ prepulse and basolateral sodium removal, respectively. Apical NHE activity was greater in 8 to 9 week old hypertensive SHR compared with WKY. Although Y-27632 suppressed pH(i) recovery in both strains, sensitivity was 50-fold higher in adult SHR. Y-27632 suppressed basolateral NHE in both strains with similar sensitivity. Apical NHE activity was not greater in 5-week-old, not yet hypertensive, SHR rats compared with WKY. In clearance studies, Na excretion was less in SHR than in WKY rats. Y-27632 increased Na excretion and fractional excretion Na in both strains but more so in SHR. (22)Na uptake of the brush border membrane vesicle taken from Y-27632-treated rats decreased more than that from vehicle-treated animals in both adult SHR and WKY. We conclude that apical NHE activity is increased in SHR PCT compared with controls and that inhibition of Rho-kinase reduces PCT NHE activities and causes natriuresis. PMID- 12538828 TI - Amifostine inhibits angiogenesis in vivo. AB - Amifostine (WR-2721) is an inorganic thiophosphate-cytoprotective agent developed to selectively protect normal tissues against the toxicity of chemotherapy and radiation. We have previously shown that amifostine protects both chicken embryo chorioallantoic membrane (CAM) vessels and cells from the effects of X-rays. In the present work, we studied the effect of amifostine on angiogenesis in vivo, using the CAM model. Amifostine decreased the number of CAM vessels in a dose dependent manner, without being toxic for the tissue. It also decreased the mRNA levels of both vascular endothelial growth factor (VEGF) isoforms VEGF(165) and VEGF(190), 6 and up to 48 h after its application onto the CAM. Similarly, it decreased the mRNA levels of inducible nitric-oxide synthase, 24 and 48 h after drug application. Furthermore, amifostine decreased the deposited amounts of laminin and collagen I 24 h after its application, without affecting the expression of the corresponding genes. The protein amounts and activity of matrix metalloproteinase-2 were not affected, whereas the expression of the corresponding gene was decreased up to 48 h after drug application. Finally, the activity of plasmin was increased 6 h after amifostine application and remained increased at later time points. These findings suggest that amifostine alters the expression of several molecules implicated in the angiogenesis process and affects the composition of the extracellular matrix in a way that leads to inhibition of angiogenesis. Such an antiangiogenic action of amifostine, together with its radioprotective effects, further supports its use in combination with radiotherapy for increased therapeutic efficacy. PMID- 12538829 TI - Influence of ovarian sex steroids on spinal methionine-enkephalin release: comparison with dynorphin reveals asymmetrical regulation. AB - The concomitant activation of spinal kappa- and delta-opioid systems is a prerequisite for the antinociception of gestation and its hormonal simulation [via 17 beta-estradiol and progesterone administration; hormone-simulated pregnancy (HSP)]. However, it is not known whether the release of kappa- and delta-opioids is also concomitantly regulated. This study investigates whether the release of methionine-enkephalin and modulation thereof is altered during HSP, as has been reported for dynorphin. K+-stimulated release of spinal methionine-enkephalin from lumbar spinal tissue obtained from control animals is negatively modulated by nociceptin (orphanin FQ; N/OFQ) in a dose-dependent manner, but not by opioids. Conversely, selective blockade of spinal N/OFQ, but not opioid receptors, augments the K+-induced increase in methionine-enkephalin release, indicating that endogenous N/OFQ also functions as a negative modulator of methionine-enkephalin release. The magnitude of K+-evoked methionine enkephalin release from spinal tissue obtained from ovarian steroid-treated animals remains unchanged, consistent with the insensitivity of its modulation by N/OFQ to the ovarian sex steroid milieu. These characteristics of methionine enkephalin release stand in sharp contrast to those previously reported for the evoked release of spinal dynorphin. Dynorphin release is subject to negative modulation by opioid (predominantly delta) as well as N/OFQ, both of which are offset during HSP, resulting in an approximately 2-fold increase in the magnitude of its release. These observations reveal that regulation of spinal dynorphin/kappa- and methionine-enkephalin/delta-spinal opioid antinociceptive systems is independent, divergent, and not symmetrical and support the formulation that spinal methionine-enkephalin/delta-opioid tone acts in a permissive/facilitative capacity to accentuate spinal dynorphin/kappa-activity. PMID- 12538830 TI - Regulation of CYP3A5 by glucocorticoids and cigarette smoke in human lung-derived cells. AB - CYP3A5 is the major CYP3A form in the human lung, and it is inducible by dexamethasone in the human A549 lung adenocarcinoma cell line. In the present study, we characterized the nature and mechanism of this induction process. The induction of CYP3A5 mRNA was assessed by quantitative reverse transcriptase polymerase chain reaction in A549 cells. About 4-fold induction was detected by nanomolar concentrations of dexamethasone and also by budenoside and beclomethasone dipropionate, glucocorticoids used for the inhalation treatment of bronchial asthma, whereas the CYP3A4 inducers mifepristone (RU486), rifampicin, clotrimazole, and nifedipine were without effect. The glucocorticoid induction was blocked by the glucocorticoid receptor (GR) antagonist RU486. In transient transfection assays to A549 cells, CYP3A5 5' regulatory region was activated by the dexamethasone treatment. In contrast, dexamethasone was unable to induce CYP3A5 transcription in GR-deficient COS-1 cells, but the induction could be achieved after GR cotransfection. The CYP3A5 expression was measured in alveolar macrophages from patients with respiratory diseases. The CYP3A5 expression level was decreased by smoking, but glucocorticoid therapy had no statistically significant effect. In conclusion, CYP3A5 is induced in the A549 cells by glucocorticoids through a GR-mediated pathway, whereas smoking may be able to depress CYP3A5 expression. PMID- 12538831 TI - Inhibition of nucleoside transport by protein kinase inhibitors. AB - Recently we reported that the pyridinylimidazole class of p38 mitogen-activated protein (MAP) kinase inhibitors potently inhibited the facilitated transport of nucleosides and nucleoside analogs in K562 cells. These compounds competed with the binding of nitrobenzylthioinosine (NBMPR) to K562 cells, consistent with inhibition of the NBMPR-sensitive equilibrative transporter (ENT1). In this study we examined a large number of additional protein kinase inhibitors for their effects on nucleoside transport. We find that incubation of K562 cells with tyrosine kinase inhibitors (AG825, AG1517, AG1478, STI-571), protein kinase C (PKC) inhibitors (staurosporine, GF 109203X, R0 31-8220, arcyriarubin A), cyclin dependent kinase inhibitors (roscovitine, olomoucine, indirubin-3'-monoxime), or rapamycin resulted in a dose-dependent reduction of intracellular uptake of [3H]uridine. In contrast, neither the MAP kinase kinase inhibitors (U0126, PD 98059) nor the phosphatidyl inositol-3 kinase inhibitors (wortmannin, LY 294002) affected this process. Furthermore, both transient uptake and prolonged [3H]thymidine incorporation in K562 cells were inhibited by protein kinase inhibitors, inactive analogs of kinase inhibitors (R0 31-6045, SB202474), and NBMPR, independently of effects on cell proliferation as determined by MTT assay. These studies demonstrate that a wide variety of protein kinase inhibitors affect nucleoside uptake through selective inhibition of nucleoside transporters, independently of kinase inhibition. PMID- 12538832 TI - Norepinephrine-induced stimulation of p38 mitogen-activated protein kinase is mediated by arachidonic acid metabolites generated by activation of cytosolic phospholipase A(2) in vascular smooth muscle cells. AB - p38 mitogen-activated protein kinase (MAPK) is activated by norepinephrine (NE) in the vasculature and is implicated in vascular smooth muscle hypertrophy, contraction, and cell migration. NE promotes influx of Ca(2+) and activates cytosolic phospholipase A(2) (cPLA(2)) in vascular smooth muscle cells (VSMC). The purpose of this study was to determine the contribution of cPLA(2)-generated arachidonic acid (AA) and its metabolites to the activation of p38 MAPK measured by its phosphorylation, in response to NE in rabbit VSMC. NE-induced p38 MAPK activation was found to be mediated through the stimulation of alpha-1 and alpha 2 adrenergic receptors, was dependent on extracellular Ca(2+), and was attenuated by an inhibitor of cPLA(2) (pyrrolidine-1). Moreover, the cPLA(2) product, AA, activated p38 MAPK in VSMC. p38 MAPK activation elicited by NE was decreased significantly by the lipoxygenase (LO) inhibitor baicalein, and to a lesser extent by the cytochrome P450 inhibitor 17-octadecynoic acid, but was not affected by the cyclooxygenase inhibitor indomethacin. The LO metabolites of AA, namely 5(S)-hydroxyeicosatetraenoic acid (HETE), 12(S)-HETE, and 15(S)-HETE and the cytochrome P450 metabolite 20-HETE, activated p38 MAPK. NE-induced p38 MAPK stimulation was found to be independent of phospholipase D (PLD) activation in rabbit VSMC. Transactivation of the epidermal growth factor receptor (EGFR) by NE also did not contribute to p38 MAPK activation. These data suggest that cPLA(2) generated AA and its LO metabolites mediate NE-induced p38 MAPK stimulation in rabbit VSMC by a mechanism that is independent of PLD and EGFR activation. PMID- 12538833 TI - Interdependent effect of P-glycoprotein-mediated drug efflux and intracellular drug binding on intracellular paclitaxel pharmacokinetics: application of computational modeling. AB - Intracellular concentration of paclitaxel is determined by the extracellular drug concentration, the level of the mdr1 P-glycoprotein (Pgp), and binding to intracellular proteins including tubulins/microtubules. The present study used a computational method to examine the effects of these factors, singly and in combination, on intracellular paclitaxel pharmacokinetics. The study was performed using our previously described intracellular pharmacokinetic model. The parameters representing Pgp-mediated drug efflux and intracellular drug binding (i.e., number of Pgp and binding sites and binding affinity) were altered systematically and used to generate computer simulations depicting the intracellular paclitaxel pharmacokinetics at clinically relevant extracellular (e.g., plasma) drug concentrations. The simulation results indicate that all four factors played a role in determining the intracellular drug accumulation. The rank order of the importance of these parameters was extracellular drug concentration >> intracellular binding capacity > intracellular binding affinity > Pgp expression. The results further showed that omission of one or more of these factors in the experimental design would lead to erroneous conclusions on the importance of other factors, as simultaneous changes in more than one parameter altered the relative importance and offset the effects of other parameters. In summary, results of the present study demonstrate the use of computational modeling to depict the effects of biological parameters such as drug efflux transporters, drug binding sites, and binding affinity on intracellular accumulation and retention of drugs that bind to cellular components. PMID- 12538834 TI - Interactions of the dipeptide ester prodrugs of acyclovir with the intestinal oligopeptide transporter: competitive inhibition of glycylsarcosine transport in human intestinal cell line-Caco-2. AB - The oligopeptide transporter may be exploited to enhance the absorption of drugs by synthesizing their dipeptide ester prodrugs, which may be recognized as its substrates. Various dipeptide esters of acyclovir (ACV), an antiviral nucleoside analog, were synthesized. Enzymatic hydrolysis and affinity of the prodrugs toward the human intestinal peptide transporter hPEPT1 were studied using the human intestinal Caco-2 cell line. Affinity studies were performed by inhibiting the uptake of [(3)H]glycylsarcosine by the prodrugs. The uptake of glycylsarcosine was found to be saturable at higher concentrations and was competitively inhibited by the prodrugs of ACV. All prodrugs except Tyr-Gly-ACV demonstrated a higher affinity (1.41-4.96 mM) toward hPEPT1 than cephalexin (8.19 +/- 2.12 mM), which was used as a positive control. Two prodrugs, Gly-Val-ACV and Val-Val-ACV, showed comparable affinity to Val-ACV, an amino acid prodrug of ACV recognized by PEPT1/PEPT2. The permeability of Gly-Val-ACV (2.99 +/- 0.59 x 10( 6) cm/s) across Caco-2 was comparable with that of Val-ACV (3.01 +/- 0.21 x 10( 6) cm/s) and was significantly inhibited (63%) in presence of glycylsarcosine. The transport of GVACV across Caco-2 was saturable at higher concentrations, and the parameters were calculated as K(m) 3.16 +/- 0.31 mM and V(max) 0.014 +/- 0.00058 nmol cm(-2) min(-1). Overall, the results suggest that the dipeptide prodrugs of ACV have a high affinity toward the intestinal oligopeptide transporter hPEPT1 and therefore seem to be promising candidates in the treatment of ocular and oral herpesvirus infections, because cornea and intestinal epithelia seem to express the oligopeptide transporters. PMID- 12538835 TI - Levodopa is toxic to dopamine neurons in an in vitro but not an in vivo model of oxidative stress. AB - Levodopa is the "gold standard" for the symptomatic treatment of Parkinson's disease (PD). There is a theoretical concern, however, that levodopa might accelerate the rate of nigral degeneration, because it undergoes oxidative metabolism and is toxic to cultured dopaminergic neurons. Most in vivo studies do not show evidence of levodopa toxicity; levodopa is not toxic to normal rodents, nonhuman primates, or humans and is not toxic to dopamine neurons in dopamine lesioned rodents or nonhuman primates in most studies. However, the potential for levodopa to be toxic in vivo has not been tested under conditions of oxidative stress such as exist in PD. To assess whether levodopa is toxic under these circumstances, we have examined the effects of levodopa on dopamine neurons in mesencephalic cultures and rat pups in which glutathione synthesis has been inhibited by L-buthionine sulfoximine. Levodopa toxicity to cultured dopaminergic neurons was enhanced by glutathione depletion and diminished by antioxidants. In contrast, treatment of neonatal rats with levodopa, administered either alone or in combination with glutathione depletion, did not cause damage to the dopamine neurons of the substantia nigra or changes in striatal levels of dopamine and its metabolites. This study provides further evidence to support the notion that although levodopa can be toxic to dopamine neurons in vitro, it is not likely to be toxic to dopamine neurons in vivo and specifically in conditions such as PD. PMID- 12538836 TI - Pharmacokinetics of 5 (and 6)-carboxy-2',7'-dichlorofluorescein and its diacetate promoiety in the liver. AB - Hepatic disposition of 5 (and 6)-carboxy-2',7'-dichlorofluorescein (CDF) and its diacetate promoiety (CDFDA) was studied in isolated perfused rat livers. Livers from Wistar wild-type and multidrug resistance-associated protein (Mrp)2 deficient (TR(-)) rats were perfused with CDF in the presence or absence of probenecid. Probenecid decreased the recovery of CDF in bile approximately 4-fold in wild-type livers (65 +/- 8% versus 15 +/- 2% of dose over 2 h). In livers from TR(-) rats, CDF was not excreted into bile and probenecid decreased perfusate CDF concentrations in a concentration-dependent manner, in part due to inhibition of Mrp3. Plasma membrane vesicles from rat Mrp2- or Mrp3-transfected Sf9 cells were used to confirm that CDF is a substrate for Mrp2 and Mrp3; probenecid inhibited the transport of CDF by Mrp2 and Mrp3 in a concentration-dependent manner. CDF uptake in collagen sandwich-cultured rat hepatocytes was temperature-dependent and saturable (K(m) = 22 +/- 10 microM; V(max) = 97 +/- 9 pmol/min/mg protein). Uptake of CDF in sandwich-cultured rat hepatocytes was impaired significantly by bromosulfophthalein, a substrate for organic anion-transporting polypeptides (Oatps), but was not modulated by specific Oatp2 or organic anion transporter (Oat) substrates. CDFDA uptake was not saturable, temperature-dependent, or impaired by inhibitors. The hydrolysis of CDFDA to CDF is mediated by basic pH and esterases in biological media. CDFDA passively diffuses into hepatocytes where it is hydrolyzed to CDF. In contrast, CDF appears to be taken up by Oatp mediated transport into rat hepatocytes and effluxed via Mrp2 into bile and via Mrp3 into sinusoidal blood. PMID- 12538837 TI - Agmatine is efficiently transported by non-neuronal monoamine transporters extraneuronal monoamine transporter (EMT) and organic cation transporter 2 (OCT2). AB - Agmatine has received considerable attention recently. Available evidence suggests that agmatine functions as a neurotransmitter and inhibits, via induction of antizyme, cellular proliferation. Because of its positive charge, agmatine will not appreciably cross cellular membranes by simple diffusion. Indeed, all physiological models require a channel or transporter protein in the plasma membrane to effect inactivation or nonexocytotic release of agmatine. However, a transport mechanism for agmatine has not been identified on a molecular level so far. In the present study, the non-neuronal monoamine transporters, organic cation transporter (OCT) 1, OCT2, and extraneuronal monoamine transporter (EMT) (gene symbols SLC22A1-A3), both from human and rat, were examined, stably expressed in 293 cells, for [(3)H]agmatine transport. Our results indicate that OCT2 and EMT, but not OCT1, efficiently translocate agmatine. The structural homolog putrescine was not accepted as substrate. Uptake of agmatine via EMT and OCT2 was saturable, with K(m) values of 1 to 2 mM. The affinity of OCT1 was 10-fold lower. Carrier-mediated efflux of agmatine was documented in a trans-stimulation experiment. Finally, uptake of agmatine increased dramatically with increasing pH. Thus, only the singly charged species of agmatine is accepted as substrate. In conclusion, both EMT and OCT2 must be considered for the control of agmatine levels in rat and human. PMID- 12538838 TI - Anxiolytic-like and antidepressant-like activities of MCL0129 (1-[(S)-2-(4 fluorophenyl)-2-(4-isopropylpiperadin-1-yl)ethyl]-4-[4-(2-methoxynaphthalen-1 yl)butyl]piperazine), a novel and potent nonpeptide antagonist of the melanocortin-4 receptor. AB - We investigated the effects of a novel melanocortin-4 (MC4) receptor antagonist,1 [(S)-2-(4-fluorophenyl)-2-(4-isopropylpiperadin-1-yl)ethyl]-4-[4-(2 methoxynaphthalen-1-yl)butyl]piperazine (MCL0129) on anxiety and depression in various rodent models. MCL0129 inhibited [(125)I][Nle(4)-D-Phe(7)]-alpha melanocyte-stimulating hormone (alpha-MSH) binding to MC4 receptor with a K(i) value of 7.9 nM, without showing affinity for MC1 and MC3 receptors. MCL0129 at 1 microM had no apparent affinity for other receptors, transporters, and ion channels related to anxiety and depression except for a moderate affinity for the sigma(1) receptor, serotonin transporter, and alpha(1)-adrenoceptor, which means that MCL0129 is selective for the MC4 receptor. MCL0129 attenuated the alpha-MSH increased cAMP formation in COS-1 cells expressing the MC4 receptor, whereas MCL0129 did not affect basal cAMP levels, thereby indicating that MCL0129 acts as an antagonist at the MC4 receptor. Swim stress markedly induced anxiogenic-like effects in both the light/dark exploration task in mice and the elevated plus maze task in rats, and MCL0129 reversed the stress-induced anxiogenic-like effects. Under nonstress conditions, MCL0129 prolonged time spent in the light area in the light/dark exploration task and suppressed marble-burying behavior. MCL0129 shortened immobility time in the forced swim test and reduced the number of escape failures in inescapable shocks in the learned helplessness test, thus indicating an antidepressant potential. In contrast, MCL0129 had negligible effects on spontaneous locomotor activity, Rotarod performance, and hexobarbital induced anesthesia. These observations indicate that MCL0129 is a potent and selective MC4 antagonist with anxiolytic- and antidepressant-like activities in various rodent models. MC4 receptor antagonists may prove effective for treating subjects with stress-related disorders such as depression and/or anxiety. PMID- 12538839 TI - Systemic and spinal analgesic activity of a delta-opioid-selective lanthionine enkephalin analog. AB - A lanthionine enkephalin derivative, Tyr-c[D-Val(L)-Gly-Phe-D-Ala(L)]-OH (DV(L)(2)DA(L)(5)LanEnk), where Val(L) and Ala(L) denote the lanthionine amino acid ends linked via a monosulfide bridge to form the lanthionine structure, was synthesized. It was found to possess selectivity for and potency at the delta versus mu opioid receptor as defined by binding studies and by its respective activity on the mouse vas deferens compared with the guinea pig ileum. The agent produced a potent analgesia after intrathecal and intraperitoneal delivery with ED(50) values being, respectively, 0.19 mucrog and 0.49 mg/kg. The effects of the agent were reversed by the delta-selective antagonist naltrindole. These analgesic actions occurred at doses that had no effect upon general behavior or motor function. These results suggest a potent delta-preferring agent suitable for development as a systemic delta opioid analgesic. PMID- 12538840 TI - The effects of the potassium channel opener minoxidil on renal electrolytes transport in the loop of henle. AB - ATP-sensitive potassium channels (K(ATP)) in the thick ascending limb of the loop of Henle play an important role in apical K(+) recycling, a mechanism essential for maintaining the activity of the Na/2Cl/K-cotransporter. We have previously demonstrated that inhibition of K(ATP) decreases Na(+) and K(+) absorption in the loop of Henle and induces diuretic and natriuretic effects. In the present study, we used renal clearance and in vivo microperfusion techniques to evaluate the effects of the K(ATP) opener minoxidil on the urinary excretion and absorption in the loop of Henle of Na(+), K(+), Ca(2+), and Mg(2+). Intravenous injection of minoxidil (1.5 mg/kg) significantly decreased fractional Na(+) (FENa) and Mg(2+) (FEMg) excretion and urine volume with a moderate decrease in blood pressure (12%) and glomerular filtration rate (15%). Urine volume decreased 63%, and FENa and FEMg decreased 58 and 37%, respectively. In contrast, K(+) and Ca(2+) excretion did not change significantly. In the microperfusion of the loop of Henle, addition of minoxidil to the perfusion fluid significantly increased fluid (J(v)), Na(+) (J(Na)), Cl(-) (J(Cl)), and K(+) (J(K)) absorption. J(v) increased 44% (from 8.32 to 11.95 nl/min), J(Na) increased 14% (from 1.96 to 2.34 nmol/min), J(Cl) increased 21% (from 1.72 to 2.08 nmol/min), and J(K) increased 57% (from 35.8 to 56.4 pmol/min). We conclude that the activation of K(ATP) leads to stimulation of Na/2Cl/K-cotransporter activity and increases the rates of Na(+), Cl(-), and K(+) absorption in the loop of Henle, an effect contributing to the antidiuretic and antinatriuretic action of this K channel opener. PMID- 12538841 TI - A novel chymase inhibitor, 2-(5-formylamino-6-oxo-2-phenyl-1,6-dihydropyrimidine 1-yl)-N-[[,4-dioxo-1-phenyl-7-(2-pyridyloxy)]2-heptyl]acetamide (NK3201), suppressed intimal hyperplasia after balloon injury. AB - In this study, we investigated whether an orally active chymase inhibitor, 2-(5 formylamino-6-oxo-2-phenyl-1,6-dihydropyrimidine-1-yl)-N-[[3,4-dioxo-1-phenyl-7 (2-pyridyloxy)]-2-heptyl]acetamide (NK3201), prevents intimal hyperplasia in carotid arteries injured by a balloon catheter in dog. Each dog was administered NK3201 (1 mg/kg per day, p.o.) or placebo beginning 5 days before balloon injury and continuing through the experiments. Four weeks after balloon injury, NK3201 did not affect the plasma renin and angiotensin-converting enzyme activities. The chymase activity was significantly increased in the injured arteries, whereas the angiotensin-converting enzyme activity was not. NK3201 significantly reduced the chymase activity in the injured arteries. The intimal area in the placebo- and NK3201-treated group and was 0.46 +/- 0.06 and 0.24 +/- 0.04 mm2, respectively, and this difference was significant. In this study, we demonstrated for the first time that a chymase inhibitor prevented the development of intimal hyperplasia in the balloon-injured arteries. PMID- 12538842 TI - Optimal structure requirements for pluronic block copolymers in modifying P glycoprotein drug efflux transporter activity in bovine brain microvessel endothelial cells. AB - Pluronic block copolymer P85 was shown to inhibit the P-glycoprotein (Pgp) drug efflux system and to increase the permeability of a broad spectrum of drugs in the blood-brain barrier (BBB). However, there is an entire series of Pluronics varying in lengths of propylene oxide and ethylene oxide and overall lipophilicity. This study identifies those structural characteristics of Pluronics required for maximal impact on drug efflux transporter activity in bovine brain microvessel endothelial cells (BBMECs). Using a wide range of block copolymers, differing in hydrophilic-lipophilic balance (HLB), this study shows that lipophilic Pluronics with intermediate length of propylene oxide block (from 30 to 60 units) and HLB <20 are the most effective at inhibiting Pgp efflux in BBMECs. The methods used included 1) cellular accumulation studies with the Pgp substrate rhodamine 123 in BBMECs to assess Pgp activity; 2) luciferin/luciferase ATP assay to evaluate changes in cellular ATP; 3) 1,6-diphenyl-1,3,5-hexatriene membrane microviscosity studies to determine alterations in membrane fluidity; and 4) Pgp ATPase assays using human Pgp-expressing membranes. Pluronics with intermediate lipophilic properties showed the strongest fluidization effect on the cell membranes along with the most efficient reduction of intracellular ATP synthesis in BBMEC monolayers. The relationship between the structure of Pluronic block copolymers and their biological response-modifying effects in BBMECs are useful for determining formulations with maximal efficacy for increasing BBB permeability. PMID- 12538843 TI - Blockade of the thrombin receptor protease-activated receptor-1 with a small molecule antagonist prevents thrombus formation and vascular occlusion in nonhuman primates. AB - Although it is well recognized that human platelet responses to alpha-thrombin are mediated by the protease-activated receptors PAR-1 and PAR-4, their role and relative importance in platelet-dependent human disease has not yet been elucidated. Because the expression profile of PARs in platelets from nonprimates differs from humans, we used cynomolgus monkeys to evaluate the role of PAR-1 in thrombosis. Based on reverse transcription-polymerase chain reaction, PAR expression in platelets from cynomolgus monkeys consisted primarily of PAR-1 and PAR-4, thereby mirroring the profile of human platelets. We probed the role of PAR-1 in a primate model of vascular injury-induced thrombosis with the selective PAR-1 antagonist (alpha S)-N-[(1S)-3-amino-1 [[(phenylmethyl)amino]carbonyl]propyl]-alpha-[[[[[1-(2,6-dichlorophenyl)methyl]-3 (1-pyrrolidinylmethyl)-1H-indazol-6-yl]amino]carbonyl]amino]-3,4 difluorobenzenepropanamide (RWJ-58259). After pretreatment with RWJ-58259 or vehicle, both carotid arteries of anesthetized monkeys were electrolytically injured and blood flow was monitored for 60 min. Time to occlusion was significantly extended after RWJ-58259 administration (27 +/- 3 to 53 +/- 8 min; p < 0.048). Vessels from three of the five treated animals remained patent. Ex vivo platelet aggregation measurements indicated complete PAR-1 inhibition, as well as an operational PAR-4 response. Immunohistochemical staining of mural thrombi with antibodies to the platelet marker CD61 and fibrinogen indicated that RWJ-58259 significantly reduced thrombus platelet deposition. Drug treatment had no effect on key hematological or coagulation parameters. Our results provide direct evidence that PAR-1 is the primary receptor that mediates alpha-thrombin's prothrombotic actions in primates and suggest that PAR-1 antagonists may have potential for the treatment of thrombotic disorders in humans. PMID- 12538844 TI - Effects of irbesartan on cloned potassium channels involved in human cardiac repolarization. AB - We studied the effects of irbesartan, a selective angiotensin II type 1 receptor antagonist, on human ether-a-go-go-related gene (HERG), KvLQT1+minK, hKv1.5, and Kv4.3 channels using the patch-clamp technique. Irbesartan exhibited a low affinity for HERG and KvLQT1+minK channels (IC(50) = 193.0 +/- 49.8 and 314.6 +/- 85.4 microM, respectively). In hKv1.5 channels, irbesartan produced two types of block, depending on the concentration tested. At 0.1 microM, irbesartan inhibited the current in a time-dependent manner (22 +/- 3.9% at +60 mV). The blockade increased steeply with channel activation increasing at more positive potentials. However, at 10 microM, irbesartan induced a time-independent blockade that occurred in the range of potentials of channel opening, reaching its maximum at approximately 0 mV, and remaining unchanged at more positive potentials (24.0 +/- 1.0% at +60 mV). In Kv4.3 currents, irbesartan produced a concentration-dependent block, which resulted in two IC(50) values (1.0 +/- 0.1 nM and 7.2 +/- 0.6 microM). At 1 microM, it inhibited the peak current and accelerated the time course of inactivation, decreasing the total charge crossing the membrane (36.6 +/- 7.8% at +50 mV). Irbesartan shifted the inactivation curve of Kv4.3 channels, the blockade increasing as the amount of inactivated channels increased. Molecular modeling was used to define energy-minimized dockings of irbesartan to hKv1.5 and HERG channels. In conclusion, irbesartan blocks Kv4.3 and hKv1.5 channels at therapeutic concentrations, whereas the blockade of HERG and KvLQT1+minK channels occurred only at supratherapeutic levels. In hKv1.5, a receptor site is apparent on each alpha-subunit of the channel, whereas in HERG channels a common binding site is present at the pore. PMID- 12538845 TI - The corticotropin-releasing factor1 receptor antagonist R121919 attenuates the behavioral and endocrine responses to stress. AB - Corticotropin-releasing factor (CRF) is the major physiological regulator of the hypothalamic-pituitary-adrenal (HPA) axis and serves to coordinate the mammalian endocrine, autonomic, and behavioral responses to stress. Considerable literature from clinical and preclinical data suggests that hypersecretion of hypothalamic and/or extrahypothalamic CRF systems is a major factor in the pathogenesis of affective and anxiety disorders. Based on this premise, a CRF(1) receptor antagonist has been hypothesized to possess anxiolytic and/or antidepressant properties. In this study, an acute dose of the lipophilic CRF(1) receptor antagonist 3-[6-(dimethylamino)-4-methyl-pyrid-3-yl]-2,5-dimethyl-N,N-dipropyl pyrazolo[2,3-a]pyrimidin-7-amine (R121919), administered i.v. to rats with surgically implanted jugular cannula 60 min before a 5-min restraint stress, dose dependently attenuated peak plasma adrenocorticopin hormone (ACTH) and corticosterone concentrations by 91 and 75%, respectively. In a second study, acute administration of R121919 reduced measures of anxiety in a rodent defensive withdrawal paradigm. R121919 dose dependently decreased latency to exit the tube, and total time spent in the tube 60 min after a single subcutaneous administration. In addition, the ACTH and corticosterone response to novelty was decreased by 82 and 97%, respectively, at the 10-mg/kg dose of R121919. In another study, this dose was associated with approximately an 85% occupancy of the CRF(1) receptor in the cortex measured 75-min postsubcutaneous injection. These data confirm that R121919 acts as a CRF(1) receptor antagonist in vivo, attenuates HPA axis responsivity, and possesses anxiolytic properties. PMID- 12538846 TI - Inhibition of histone deacetylases by chlamydocin induces apoptosis and proteasome-mediated degradation of survivin. AB - The naturally occurring cyclic tetrapeptide chlamydocin is a very potent inhibitor of cell proliferation. Here we show that chlamydocin is a highly potent histone deacetylase (HDAC) inhibitor, inhibiting HDAC activity in vitro with an IC(50) of 1.3 nM. Like other HDAC inhibitors, chlamydocin induces the accumulation of hyperacetylated histones H3 and H4 in A2780 ovarian cancer cells, increases the expression of p21(cip1/waf1), and causes an accumulation of cells in G(2)/M phase of the cell cycle. In addition, chlamydocin induces apoptosis by activating caspase-3, which in turn leads to the cleavage of p21(cip1/waf1) into a 15-kDa breakdown product and drives cells from growth arrest into apoptosis. Concomitant with the activation of caspase-3 and cleavage of p21(cip1/waf1), chlamydocin decreases the protein level of survivin, a member of the inhibitor of apoptosis protein family that is selectively expressed in tumors. Although our data indicate a potential link between degradation of survivin and activation of the apoptotic pathway induced by HDAC inhibitors, stable overexpression of survivin does not suppress the activation of caspase-3 or cleavage of p21(cip1/waf1) induced by chlamydocin treatment. The decrease of survivin protein level is mediated by degradation via proteasomes since it can be inhibited by specific proteasome inhibitors. Taken together, our results show that induction of apoptosis by chlamydocin involves caspase-dependent cleavage of p21(cip1/waf1), which is strikingly associated with proteasome-mediated degradation of survivin. PMID- 12538847 TI - The effect of endotoxin and dexamethasone on enrofloxacin pharmacokinetic parameters in swine. AB - The impact of Escherichia coli-derived lipopolysaccharide (LPS) on the pharmacokinetic parameters of enrofloxacin in swine was assessed to determine whether this model would substitute for a pleuropneumonia infection model for pharmacokinetic evaluation of drugs. All animals received a single i.v. dose of enrofloxacin (5 mg/kg). Half the animals also received dexamethasone (0.5 mg/kg) to determine the impact of inflammation on any changes in enrofloxacin pharmacokinetics, as most of the effects of LPS are due to elaboration of inflammatory mediators. Administration of LPS alone (2.0 microg/kg) was associated with a decrease in clearance of enrofloxacin. Volume of distribution at steady state was increased in the dexamethasone-treated animals. The terminal elimination half-life of enrofloxacin was significantly increased in the LPS group. Dexamethasone administration, either alone or in combination with LPS challenge, increased the volume of distribution both at steady state and during the elimination phase. Lipopolysaccharide challenge did not affect the volume of distribution. Lipopolysaccharide challenge did not affect urinary excretion of enrofloxacin but did increase the urinary excretion of its principal metabolite, ciprofloxacin. However, the increased excretion did not begin until 24 h after administration of enrofloxacin. Because these pharamcokinetic results are different from those obtained with the pleuropneumonia model using the bacteria Actinobacillus pleuropneumoniae, the results of this study demonstrate that LPS is not a generic substitute for infection for the pharmacokinetic evaluation of drugs. PMID- 12538848 TI - Discovery of the breast cancer gene BASE using a molecular approach to enrich for genes encoding membrane and secreted proteins. AB - To identify unknown membrane proteins that could be used as targets for breast and prostate cancer immunotherapies and secreted proteins to be used as diagnostic markers, a cDNA library was generated from membrane-associated polyribosomal RNA derived from four breast cancer cell lines, one normal breast cell line, and a prostate cancer cell line. The membrane-associated polyribosomal cDNA library was subtracted with RNA from normal brain, liver, lung, kidney, and muscle. Of the 15,581 clones sequenced from the subtracted cDNA library, sequences from 10,506 clones map to known genes, but 5,075 sequences, representing 3,181 unique transcripts, are not associated with known genes. As one example, we experimentally investigated expression of a previously uncharacterized breast cancer gene that encodes a secreted protein designated BASE (breast cancer and salivary gland expression). BASE is expressed in many breast cancers but not in essential normal tissues including the five organs used for subtraction. Further analysis of this library should yield additional gene products of use in the diagnosis or treatment of breast or prostate cancer. PMID- 12538849 TI - Hair-bundle movements elicited by transepithelial electrical stimulation of hair cells in the sacculus of the bullfrog. AB - Electrically evoked otoacoustic emission is a manifestation of reverse transduction by the inner ear. We present evidence for a single-cell correlate of this phenomenon, hair-bundle movement driven by transepithelial electrical stimulation of the frog's sacculus. Responses could be observed at stimulus frequencies up to 1 kHz, an order of magnitude higher than the organ's natural range of sensitivity to acceleration or sound. Measurements at high-stimulus frequencies and pharmacological treatments allow us to distinguish two mechanisms that mediate the electrical responses: myosin-based adaptation and Ca(2+) dependent reclosure of transduction channels. These mechanisms also participate in the active process that amplifies and tunes the mechanical responses of this receptor organ. Transient application of the channel blocker gentamicin demonstrated the crucial role of mechanoelectrical transduction channels in the rapid responses to electrical stimulation. A model for electrically driven bundle motion that incorporates the negative stiffness of the hair bundle as well as its two mechanisms of motility captures the essential features of the measured responses. PMID- 12538850 TI - Emergence and evolution of Vibrio cholerae O139. AB - The emergence of Vibrio cholerae O139 Bengal during 1992-1993 was associated with large epidemics of cholera in India and Bangladesh and, initially, with a total displacement of the existing V. cholerae O1 strains. However, the O1 strains reemerged in 1994 and initiated a series of disappearance and reemergence of either of the two serogroups that was associated with temporal genetic and phenotypic changes sustained by the strains. Since the initial emergence of the O139 vibrios, new variants of the pathogen derived from multiple progenitors have been isolated and characterized. The clinical and epidemiological characteristics of these strains have been studied. Rapid genetic reassortment in O139 strains appears to be a response to the changing epidemiology of V. cholerae O1 and also a strategy for persistence in competition with strains of the O1 serogroup. The emergence of V. cholerae O139 has provided a unique opportunity to witness genetic changes in V. cholerae that may be associated with displacement of an existing serogroup by a newly emerging one and, thus, provide new insights into the epidemiology of cholera. The genetic changes and natural selection involving both environmental and host factors are likely to influence profoundly the genetics, epidemiology, and evolution of toxigenic V. cholerae, not only in the Ganges Delta region of India and Bangladesh, but also in other areas of endemic and epidemic cholera. PMID- 12538851 TI - The hippocampus mediates glucocorticoid-induced impairment of spatial memory retrieval: dependence on the basolateral amygdala. AB - Previous studies have indicated that stress-activated glucocorticoid hormones induce temporary memory retrieval impairment. The present study examined whether adrenal steroid receptors in the hippocampus mediate such glucocorticoid effects on spatial memory retrieval. The specific glucocorticoid receptor (GR) agonist 11beta, 17beta-dihydroxy-6,21-dimethyl-17alpha-pregna-4,6-trien-20yn-3-one (RU 28362; 5 or 15 ng) infused into the hippocampus of male Sprague-Dawley rats 60 min before water-maze retention testing, 24 h after training, dose-dependently impaired probe-trial retention performance, as assessed both by time spent in the training quadrant and initial latency to cross the platform location. The GR agonist did not affect circulating corticosterone levels immediately after the probe trial, indicating that RU 28362 infusions did not influence retention by altering glucocorticoid feedback mechanisms. As infusions of the GR agonist into the hippocampus 60 min before training did not influence water-maze acquisition or immediate recall, the findings indicated that the GR agonist-induced retention impairment was induced selectively by an influence on information retrieval. In contrast, pretest infusions of the GR agonist administered into the basolateral complex of the amygdala (BLA; 2 or 6 ng) did not alter retention performance in the water maze. However, N-methyl-d-aspartate-induced lesions of the BLA, made 1 week before training, blocked the memory retrieval impairment induced by intrahippocampal infusions of RU 28362 given 60 min before the retention test. These findings indicate that the effects of glucocorticoids on retrieval of long term spatial memory depend on the hippocampus and, additionally, that neuronal input from the BLA is critical in enabling hippocampal glucocorticoid effects on memory retrieval. PMID- 12538852 TI - Growth hormone-releasing hormone (GHRH) antagonists inhibit the proliferation of androgen-dependent and -independent prostate cancers. AB - The antiproliferative effects of an antagonist of growth hormone-releasing hormone (GHRH) JV-1-38 were evaluated in nude mice bearing s.c. xenografts of LNCaP and MDA-PCa-2b human androgen-sensitive and DU-145 androgen-independent prostate cancers. In the androgen-sensitive models, JV-1-38 greatly potentiated the antitumor effect of androgen deprivation induced by surgical castration, but was ineffective when given alone. Thus, in castrated animals bearing MDA-PCa-2b cancers, the administration of JV-1-38 for 35 days virtually arrested tumor growth (94% inhibition vs. intact control, P < 0.01; and 75% vs. castrated control, P < 0.05). The growth of LNCaP tumors was also powerfully suppressed by JV-1-38 combined with castration (83% inhibition vs. intact control, P < 0.01; and 68% vs. castrated control, P < 0.05). However, in androgen-independent DU-145 cancers, JV-1-38 alone could inhibit tumor growth by 57% (P < 0.05) after 45 days. In animals bearing MDA-PCa-2b and LNCaP tumors, the reduction in serum prostate-specific antigen levels, after therapy with JV-1-38, paralleled the decrease in tumor volume. Inhibition of MDA-PCa-2b and DU-145 cancers was associated with the reduction in the expression of mRNA and protein levels of vascular endothelial growth factor. The mRNA expression for GHRH receptor splice variants was found in all these models of prostate cancer. Our results demonstrate that GHRH antagonists inhibit androgen-independent prostate cancers and, after combination with androgen deprivation, also androgen-sensitive tumors. Thus, the therapy with GHRH antagonist could be considered for the management of both androgen-dependent or -independent prostate cancers. PMID- 12538853 TI - Secretory granules are recaptured largely intact after stimulated exocytosis in cultured endocrine cells. AB - Classical cell biology teaches that exocytosis causes the membrane of exocytic vesicles to disperse into the cell surface and that a cell must later retrieve by molecular sorting whatever membrane components it wishes to keep inside. We have tested whether this view applies to secretory granules in intact PC-12 cells. Three granule proteins were labeled with fluorescent proteins in different colors, and two-color evanescent-field microscopy was used to view single granules during and after exocytosis. Whereas neuro-peptide Y was lost from granules in seconds, tissue plasminogen activator (tPA) and the membrane protein phogrin remained at the granule site for over 1 min, thus providing markers for postexocytic granules. When tPA was imaged simultaneously with cyan fluorescent protein (CFP) as a cytosolic marker, the volume occupied by the granule appeared as a dark spot where it excluded CFP. The spot remained even after tPA reported exocytosis, indicating that granules failed to flatten into the cell surface. Phogrin was labeled with GFP at its luminal end and used to sense the pH in granules. When exocytosis caused the acidic granule interior to neutralize, GFP phogrin at first brightened and later dimmed again as the interior separated from the extracellular space and reacidified. Reacidification and dimming could be reversed by application of NH(4)Cl. We conclude that most granules reseal in <10 s after releasing cargo, and that these empty or partially empty granules are recaptured otherwise intact. PMID- 12538854 TI - Suppression of cortical representation through backward conditioning. AB - Temporal stimulus reinforcement sequences have been shown to determine the directions of synaptic plasticity and behavioral learning. Here, we examined whether they also control the direction of cortical reorganization. Pairing ventral tegmental area stimulation with a sound in a backward conditioning paradigm specifically reduced representations of the paired sound in the primary auditory cortex (AI). This temporal sequence-dependent bidirectional cortical plasticity modulated by dopamine release hypothetically serves to prevent the over-representation of frequently occurring stimuli resulting from their random pairing with unrelated rewards. PMID- 12538855 TI - Lectin-like oxidized LDL receptor-1 is a cell-adhesion molecule involved in endotoxin-induced inflammation. AB - Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is a major endothelial receptor for oxidized low-density lipoprotein, and is assumed to play a proatherogenic role in atherosclerosis. LOX-1 expression is induced by inflammatory cytokines as well as by proatherogenic stimuli. LOX-1 protein binds agedapoptotic cells, activated platelets, and bacteria, suggesting that it may have diverse activities in vivo. Here, we reveal a role for LOX-1 in endotoxin induced inflammation. In a model of endotoxemia, injection of a high dose of endotoxin into rats induced leukopenia within 1 h and death of the animals within 24 h. Preadministration of anti-LOX-1 antibody reduced the degree of leukopenia and completely rescued the animals, whereas control IgG did not. In a model of low-dose endotoxin-induced uveitis, anti-LOX-1 antibody efficiently suppressed leukocyte infiltration and protein exudation. In situ videomicroscopic analyses of leukocyte interactions with retinal veins revealed that anti-LOX-1 antibody reduced the number of rolling leukocytes and increased the velocity of rolling, suggesting that LOX-1 functions as a vascular tethering ligand. The ability of LOX-1 to capture leukocytes under physiologic shear was confirmed in an in vitro flow model. Thus, LOX-1 is an adhesion molecule involved in leukocyte recruitment and may represent an attractive target for modulation of endotoxin-induced inflammation. PMID- 12538856 TI - Interactions among three distinct CesA proteins essential for cellulose synthesis. AB - In a screen to identify novel cellulose deficient mutants, three lines were shown to be allelic and define a novel complementation group, irregular xylem5 (irx5). IRX5 was cloned and encodes a member of the CesA family of cellulose synthase catalytic subunits (AtCesA4). irx5 plants have an identical phenotype to previously described mutations in two other members of this gene family (IRX1 and IRX3). IRX5, IRX3, and IRX1 are coexpressed in exactly the same cells, and all three proteins interact in detergent solubilized extracts, suggesting that three members of this gene family are required for cellulose synthesis in secondary cell walls. The association of IRX1 and IRX3 was reduced to undetectable levels in the absence of IRX5. Consequently, these data suggest that IRX5, IRX3, and IRX1 are all essential components of the cellulose synthesizing complex and the presence of all three subunits is required for the correct assembly of this complex. PMID- 12538857 TI - Requirement of the calcineurin subunit gene canB2 for indirect flight muscle formation in Drosophila. AB - Calcineurin is a calcium-activated protein phosphatase involved in multiple aspects of cardiac and skeletal muscle development and disease. Genes encoding calcineurin subunit proteins are highly conserved among animal species. Toward the goal of identifying new calcineurin-interacting loci that function in myogenic processes, we expressed an activated form of mouse calcineurin A in Drosophila and screened for suppressors of the phosphatase-induced lethality. Here, we demonstrate that a mutation in the canB2 gene, which encodes a regulatory subunit of Drosophila calcineurin, can suppress a pupal developmental arrest phenotype to adult viability. As canB2 is an essential gene and rare homozygous escapers are flightless, we further analyzed canB2 expression and function in pupae and adults. The gene is expressed in the forming indirect flight muscles and central nervous system during pupal development. A canA gene is comparably expressed in these tissues. Consistent with the observed muscle expression, canB2 mutants exhibit severe defects in the organization of their indirect flight muscles, a phenotype that is likely caused by muscle hypercontractility. Together, these findings demonstrate a vital role for the phosphatase in this specific facet of Drosophila myogenesis and show conserved fly and vertebrate calcineurin genes contribute prominently to fundamental processes of muscle formation and function. PMID- 12538858 TI - The role of nuclear factor-kappaB essential modulator (NEMO) in B cell development and survival. AB - The transcription factor nuclear factor-kappaB (NF-kappaB) is essential for immune and inflammatory responses. NF-kappaB essential modulator (NEMO) is a scaffolding component of the IkappaB kinase complex required for NF-kappaB activation in vitro. Because NF-kappaB activation is involved in B cell development and function, we set out to determine whether NEMO is required for these processes. NEMO(-/-) mice die very early during embryogenesis, and fetal livers from NEMO(-/-) embryos can not reconstitute either B or T lymphopoiesis in irradiated host mice. We therefore used NEMO(-/-) embryonic stem cells and the OP9 in vitro differentiation system to demonstrate that NEMO is not required for B cell development but plays an important role in B cell survival. PMID- 12538859 TI - Strikingly higher frequency in centenarians and twins of mtDNA mutation causing remodeling of replication origin in leukocytes. AB - The presence of a genetic component in longevity is well known. Here, the association of a mtDNA mutation with a prolonged life span in humans was investigated. Large-scale screening of the mtDNA main control region in leukocytes from subjects of an Italian population revealed a homoplasmic C150T transition near an origin of heavy mtDNA-strand synthesis in approximately 17% of 52 subjects 99-106 years old, but, in contrast, in only 3.4% of 117 younger individuals (P = 0.0035). Evidence was obtained for the contribution of somatic events, under probable nuclear genetic control, to the striking selective accumulation of the mutation in centenarians. In another study, among leukocyte mtDNA samples from 20 monozygotic and 18 dizygotic twins, 60-75 years old, 30% (P = 0.0007) and 22% (P = 0.011), respectively, of the individuals involved exhibited the homoplasmic C150T mutation. In a different system, i.e., in five human fibroblast longitudinal studies, convincing evidence for the aging-related somatic expansion of the C150T mutation, up to homoplasmy, was obtained. Most significantly, 5' end analysis of nascent heavy mtDNA strands consistently revealed a new replication origin at position 149, substituting for that at 151, only in C150T mutation-carrying samples of fibroblasts or immortalized lymphocytes. Considering the aging-related health risks that the centenarians have survived and the developmental risks of twin gestations, it is proposed that selection for a remodeled replication origin, inherited or somatically acquired, provides a survival advantage and underlies the observed high incidence of the C150T mutation in centenarians and twins. PMID- 12538860 TI - A single-stranded promoter for RNA polymerase III. AB - Single strands of DNA serve, in rare instances, as promoters for transcription; duplex DNA promoters with individual strands that also have a promoter capacityfunction have not been described. We show that the nontranscribed strand of the Saccharomyces cerevisiae U6 snRNA gene directs transcription initiation factor IIIB-requiring and accurately initiating transcription by RNA polymerase III. The nontranscribed strand promoter is much more extended than its duplex DNA counterpart, comprising the U6 gene TATA box, a downstream T(7) tract, and an upstream-lying segment. A requirement for placement of the 3' end of the transcribed (template) strand within the confines of the transcription bubble is seen as indicating that the nontranscribed strand provides a scaffold for RNA polymerase recruitment but is deficient at a subsequent step of transcription initiation factor IIIB's direct involvement in promoter opening. PMID- 12538861 TI - Oncogenic targeting of an activated tyrosine kinase to the Golgi apparatus in a glioblastoma. AB - Activating oncogenic mutations of receptor tyrosine kinases (RTKs) have been reported in several types of cancers. In many cases, genomic rearrangements lead to the fusion of unrelated genes to the DNA coding for the kinase domain of RTKs. All RTK-derived fusion proteins reported so far display oligomerization sequences within the 5' fusion partners that are responsible for oncogenic activation. Here, we report a mechanism by which an altered RTK gains oncogenic potential in a glioblastoma cell line. A microdeletion on 6q21 results in the fusion of FIG, a gene coding for a Golgi apparatus-associated protein, to the kinase domain of the protooncogene c-ROS. The fused protein product FIG-ROS is a potent oncogene, and its transforming potential resides in its ability to interact with and become localized to the Golgi apparatus. Thus we have found a RTK fusion protein whose subcellular location leads to constitutive kinase activation and results in oncogenic transformation. PMID- 12538862 TI - Adenosine receptor blockade reverses hypophagia and enhances locomotor activity of dopamine-deficient mice. AB - Adenosine receptors modulate dopaminergic function by regulating dopamine release in presynaptic neurons and intracellular signaling in postsynaptic striatal neurons. To investigate how adenosine impinges on the action of dopamine in feeding and locomotion, genetically altered, dopamine-deficient mice were treated with adenosine receptor antagonists. Acute administration of the nonselective adenosine receptor antagonist, caffeine (5-25 mgkg i.p.), reversed the hypophagia of mutant mice and induced hyperactivity in both control and mutant animals. However, caffeine treatment elicited much less hyperactivity in dopamine deficient mice than did l-3,4-dihydroxyphenylalanine (l-dopa) administration, which partially restores dopamine content. Caffeine treatment enhanced feeding of l-dopa-treated mutants but, unexpectedly, it reduced their hyperlocomotion. Caffeine administration induced c-Fos expression in the cortex of dopamine deficient mice but had no effect in the striatum by itself. Caffeine attenuated dopamine agonist-induced striatal c-Fos expression. An antagonist selective for adenosine A(2A) receptors induced feeding and locomotion in mutants much more effectively than an A(1) receptor antagonist. l-dopa-elicited feeding and hyperlocomotion were reduced in mutants treated with an A(1) receptor agonist, whereas an A(2A) receptor agonist decreased l-dopa-induced feeding without affecting locomotion. The observations suggest that the hypophagia and hypoactivity of mutants result not only because of the absence of dopamine but also because of the presence of A(2A) receptor signaling. This study of a genetic model of dopamine depletion provides evidence that A(2A) receptor antagonists could ameliorate the hypokinetic symptoms of advanced Parkinson's disease patients without inducing excessive motor activity. PMID- 12538863 TI - Biochemical characterization of the Yersinia YopT protease: cleavage site and recognition elements in Rho GTPases. AB - The Gram-negative bacterial pathogen Yersinia delivers six effector proteins into the host cells to thwart the host innate immune response. One of the effectors, YopT, causes the disruption of the actin cytoskeleton and contributes to the inhibition of phagocytosis of the pathogen. YopT functions as a cysteine protease to cleave Rho family GTPases. We have analyzed the YopT cleavage products of Rho GTPases by TLC and determined their chemical structure by MS. Amino acid labeling experiments were performed to locate the exact site in RhoA where the YopT cleavage occurs. Our data unambiguously demonstrate that YopT cleaves N-terminal to the prenylated cysteine in RhoA, Rac, and Cdc42 and that the cleavage product of the GTPases is geranylgeranyl cysteine methyl ester. YopT cleaves GTP- and GDP bound forms of RhoA equally, suggesting that the cleavage does not depend upon the conformation status of the GTPases. YopT also cleaves both farnesylated and geranylgeranylated forms of RhoA. The polybasic sequence in the C terminus of RhoA is essential for YopT substrate recognition and cleavage. PMID- 12538864 TI - Transplanted bone marrow generates new neurons in human brains. AB - Adult bone marrow stem cells seem to differentiate into muscle, skin, liver, lung, and neuronal cells in rodents and have been shown to regenerate myocardium, hepatocytes, and skin and gastrointestinal epithelium in humans. Because we have demonstrated previously that transplanted bone marrow cells can enter the brain of mice and differentiate into neurons there, we decided to examine postmortem brain samples from females who had received bone marrow transplants from male donors. The underlying diseases of the patients were lymphocytic leukemia and genetic deficiency of the immune system, and they survived between 1 and 9 months after transplant. We used a combination of immunocytochemistry (utilizing neuron specific antibodies) and fluorescent in situ hybridization histochemistry to search for Y chromosome-positive cells. In all four patients studied we found cells containing Y chromosomes in several brain regions. Most of them were nonneuronal (endothelial cells and cells in the white matter), but neurons were certainly labeled, especially in the hippocampus and cerebral cortex. The youngest patient (2 years old), who also lived the longest time after transplantation, had the greatest number of donor-derived neurons (7 in 10,000). The distribution of the labeled cells was not homogeneous. There were clusters of Y-positive cells, suggesting that single progenitor cells underwent clonal expansion and differentiation. We conclude that adult human bone marrow cells can enter the brain and generate neurons just as rodent cells do. Perhaps this phenomenon could be exploited to prevent the development or progression of neurodegenerative diseases or to repair tissue damaged by infarction or trauma. PMID- 12538865 TI - A quantitative structure-function relationship for the Photosystem II reaction center: supermolecular behavior in natural photosynthesis. AB - Light-induced charge separation is the primary photochemical event of photosynthesis. Efficient charge separation in photosynthetic reaction centers requires the balancing of electron and excitation energy transfer processes, and in Photosystem II (PSII), these processes are particularly closely entangled. Calculations that treat the cofactors of the PSII reaction center as a supermolecular complex allow energy and electron transfer reactions to be described in a unified way. This calculational approach is shown to be in good agreement with experimentally observed energy and electron transfer dynamics. This supermolecular view also correctly predicts the effect of changing the redox potentials of cofactors by site-directed mutagenesis, thus providing a unified and quantitative structure-function relationship for the PSII reaction center. PMID- 12538866 TI - Structure of the large FK506-binding protein FKBP51, an Hsp90-binding protein and a component of steroid receptor complexes. AB - The ability to bind immunosuppressive drugs such as cyclosporin and FK506 defines the immunophilin family of proteins, and the FK506-binding proteins form the FKBP subfamily of immunophilins. Some FKBPs, notably FKBP12 (the 12-kDa FK506-binding protein), have defined roles in regulating ion channels or cell signaling, and well established structures. Other FKBPs, especially the larger ones, participate in important biological processes, but their exact roles and the structural bases for these roles are poorly defined. FKBP51 (the 51-kDa FKBP) associates with heat shock protein 90 (Hsp90) and appears in functionally mature steroid receptor complexes. In New World monkeys, FKBP51 has been implicated in cortisol resistance. We report here the x-ray structures of human FKBP51, to 2.7 A, and squirrel monkey FKBP51, to 2.8 A, by using multiwavelength anomalous dispersion phasing. FKBP51 is composed of three domains: two consecutive FKBP domains and a three-unit repeat of the TPR (tetratricopeptide repeat) domain. This structure of a multi-FKBP domain protein clarifies the arrangement of these domains and their possible interactions with other proteins. The two FKBP domains differ by an insertion in the second that affects the formation of the progesterone receptor complex. PMID- 12538867 TI - Stabilization of perfect and imperfect tandem repeats by single-strand DNA exonucleases. AB - Rearrangements between tandemly repeated DNA sequences are a common source of genetic instability. Such rearrangements underlie several human genetic diseases. In many organisms, the mismatch-repair (MMR) system functions to stabilize repeats when the repeat unit is short or when sequence imperfections are present between the repeats. We show here that the action of single-stranded DNA (ssDNA) exonucleases plays an additional, important role in stabilizing tandem repeats, independent of their role in MMR. For perfect repeats of approximately 100 bp in Escherichia coli that are not susceptible to MMR, exonuclease (Exo)-I, ExoX, and RecJ exonuclease redundantly inhibit deletion. Our data suggest that >90% of potential deletion events are avoided by the combined action of these three exonucleases. Imperfect tandem repeats, less prone to rearrangements, are stabilized by both the MMR-pathway and ssDNA-specific exonucleases. For 100-bp repeats containing four mispairs, ExoI alone aborts most deletion events, even in the presence of a functional MMR system. By genetic analysis, we show that the inhibitory effect of ssDNA exonucleases on deletion formation is independent of the MutS and UvrD proteins. Exonuclease degradation of DNA displaced during the deletion process may abort slipped misalignment. Exonuclease action is therefore a significant force in genetic stabilization of many forms of repetitive DNA. PMID- 12538868 TI - Evaporation and instabilities of microscopic capillary bridges. AB - The formation and disappearance of liquid bridges between two surfaces can occur either through equilibrium or nonequilibrium processes. In the first instance, the bridge molecules are in thermodynamic equilibrium with the surrounding vapor medium. In the second, chemical potential gradients result in material transfer; mechanical instabilities, because of van der Waals force jumps on approach or a Rayleigh instability on rapid separation, may trigger irreversible film coalescence or bridge snapping. We have studied the growth and disappearance mechanisms of laterally microscopic liquid bridges of three hydrocarbon liquids in slit-like pores. At rapid slit-opening rates, the bridges rupture by means of a mechanical instability described by the Young-Laplace equation. Noncontinuum but apparently reversible behavior is observed when a bridge is held at nanoscopic surface separations H close to the thermodynamic equilibrium Kelvin length, 2r(K)costheta, where r(K) is the Kelvin radius and theta is the contact angle. During the course of slow evaporation (at H > 2r(K)costheta) and subsequent regrowth by capillary condensation (at H < 2r(K)costheta), the refractive index of the bridge may vary continuously and reversibly between that of the bulk liquid and vapor. The evaporation process becomes irreversible only at the very final stage of evaporation, when the refractive index of the fluid attains virtually that of the vapor. Measured refractive index profiles and the time-dependence of evaporating neck diameters also seem to differ from predictions based on a continuum picture of bridge evaporation far from the critical point. We discuss these findings in terms of the probable density profiles in evolving liquid bridges. PMID- 12538869 TI - Genetic diversity, asymmetrical aggression, and recognition in a widespread invasive species. AB - The evolution and persistence of cooperative social units depends on the ability to distinguish group members from nonmembers. The precision of discrimination, in turn, relies on variation in the labels that individuals use to recognize group members. However, this same variation can be selected against if individuals that are rejected as nonmembers incur a high cost. Here we provide evidence that selection against individuals from genetically diverse groups has contributed to the formation of the unicolonial colony structure that characterizes introduced populations of the invasive Argentine ant (Linepithema humile). Studies in both the laboratory and the field showed that individuals from less genetically diverse colonies attack individuals from more diverse colonies and that attackers survived agonistic encounters more than six times as often as recipients of aggression. This selection, in concert with reductions in genetic diversity after a founder event, likely creates a barrier to the establishment of new, genetically diverse introductions from the native range and may reduce genetic diversity within established populations in the introduced range. PMID- 12538870 TI - Crystal structure of the MurG:UDP-GlcNAc complex reveals common structural principles of a superfamily of glycosyltransferases. AB - MurG is an essential glycosyltransferase that forms the glycosidic linkage between N-acetyl muramyl pentapeptide and N-acetyl glucosamine in the biosynthesis of the bacterial cell wall. This enzyme is a member of a major superfamily of NDP-glycosyltransferases for which no x-ray structures containing intact substrates have been reported. Here we present the 2.5-A crystal structure of Escherichia coli MurG in complex with its donor substrate, UDP-GlcNAc. Combined with genomic analysis of other superfamily members and site-specific mutagenesis of E. coli MurG, this structure sheds light on the molecular basis for both donor and acceptor selectivity for the superfamily. This structural analysis suggests that it will be possible to evolve new glycosyltransferases from prototypical superfamily members by varying two key loops while maintaining the overall architecture of the family and preserving key residues. PMID- 12538871 TI - Metabotropic mGlu5 receptors regulate adenosine A2A receptor signaling. AB - Dopamine, by activating dopamine D1-type receptors, and adenosine, by activating adenosine A(2A) receptors, stimulate phosphorylation of DARPP-32 (dopamine- and cAMP-regulated phosphoprotein of M(r) 32,000) at Thr-34. In this study, we investigated the effect of metabotropic glutamate (mGlu) receptors on DARPP-32 phosphorylation at Thr-34 in neostriatal slices. A broad-spectrum mGlu receptor agonist, trans-ACPD, and a group I mGlu receptor agonist, DHPG, stimulated DARPP 32 phosphorylation at Thr-34. Studies with mGlu receptor antagonists revealed that the effects of trans-ACPD and DHPG were mediated through activation of mGlu5 receptors. The action of mGlu5 receptors required activation of adenosine A(2A) receptors by endogenous adenosine. Conversely, the action of adenosine A(2A) receptors required activation of mGlu5 receptors by endogenous glutamate. Coactivation of mGlu5 and adenosine A(2A) receptors by exogenous agonists synergistically increased DARPP-32 phosphorylation. mGlu5 receptors did not require activation of dopamine D1-type receptors by endogenous dopamine, nor did dopamine D1-type receptors require activation of mGlu5 receptors by endogenous glutamate. DHPG potentiated the effect of forskolin, but not that of 8-bromo cAMP, and stimulated DARPP-32 phosphorylation in the presence of the phosphodiesterase inhibitor IBMX, suggesting that mGlu5 receptors stimulate the rate of cAMP formation coupled to adenosine A(2A) receptors. The action of mGlu5 receptors was attenuated by inhibitors of extracellular signal-regulated kinase, but not by inhibitors of phospholipase C, p38, casein kinase 1, or Cdk5. The results demonstrate that mGlu5 receptors potentiate adenosine A(2A)DARPP-32 signaling by stimulating the adenosine A(2A) receptor-mediated formation of cAMP in an extracellular signal-regulated kinase-dependent manner. PMID- 12538872 TI - The mouse organellar biogenesis mutant buff results from a mutation in Vps33a, a homologue of yeast vps33 and Drosophila carnation. AB - In the mouse, more than 16 loci are associated with mutant phenotypes that include defective pigmentation, aberrant targeting of lysosomal enzymes, prolonged bleeding, and immunodeficiency, the result of defective biogenesis of cytoplasmic organelles: melanosomes, lysosomes, and various storage granules. Many of these mouse mutants are homologous to the human Hermansky-Pudlak syndrome (HPS), Chediak-Higashi syndrome, and Griscelli syndrome. We have mapped and positionally cloned one of these mouse loci, buff (bf), which has a mutant phenotype similar to that of human HPS. Mouse bf results from a mutation in Vps33a and thus is homologous to the yeast vacuolar protein-sorting mutant vps33 and Drosophila carnation (car). This is the first found defect of the class C vacuole/prevacuole-associated target soluble N-ethylmaleimide-sensitive factor attachment protein receptor (t-SNARE) complex in mammals and the first mammalian mutant found that is directly homologous to a vps mutation of yeast. VPS33A thus is a good candidate gene for a previously uncharacterized form of human HPS. PMID- 12538873 TI - Without peripheral interference, thymic deletion is mediated in a cohort of double-positive cells without classical activation. AB - Peripheral activation can cause bystander thymocyte death by eliciting a "cytokine storm." This event complicates in vivo studies using exogenous ligand induced models of negative selection. A stable transgenic model that selectively eliminates peripheral CD4 cells has allowed us to analyze negative selection as direct cognate events in two T cell receptor transgenic mice, OT-II and DO11. Whereas cognate peptide induced a massive deletion in double-positive (DP) cells in mice with peripheral CD4 cells, this DP deletion was modest in mice lacking peripheral CD4 cells. Using BrdUrd and annexin V staining, we found that negative selection primarily occurs in a cohort of DP cells and the absence of single positive (SP) cells is largely caused by reduction in the cohort of DP precursors. Moreover, the fates of DP cells and SP cells after antigen exposure were vastly different. Whereas SP cells up-regulated uniformly their CD69 and CD44 levels, increased their cell size, and survived after antigen exposure, DP cells had less CD69 and CD44 up-regulation, no size change, and promptly died. Thus, negative selection represents an "abortive" activation different from activation-induced cell death of mature T cells. PMID- 12538874 TI - Sodium ion cycling mediates energy coupling between complex I and ATP synthase. AB - We show here sodium ion cycling between complex I from Klebsiella pneumoniae and the F(1)F(0) ATP synthase from Ilyobacter tartaricus in a reconstituted proteoliposome system. In the course of NADH oxidation by complex I, an electrochemical sodium ion gradient was established and served as a driving force for the synthesis of ATP from ADP and phosphate. In the opposite direction, the deltamu(Na(+)) generated by ATP hydrolysis could be coupled to NADH formation by reversed electron transfer from ubiquinol to NAD. For reverse electron transfer, a transmembrane voltage larger than 30 mV was obligatory. No NADH-driven proton transport into the lumen of proteoliposomes was detected. We conclude that Na(+) is used as the exclusive coupling ion by the enterobacterial complex I. PMID- 12538875 TI - Modular organization of cellular networks. AB - We investigated the organization of interacting proteins and protein complexes into networks of modules. A network-clustering method was developed to identify modules. This method of network-structure determination was validated by clustering known signaling-protein modules and by identifying module rudiments in exclusively high-throughput protein-interaction data with high error frequencies and low coverage. The signaling network controlling the yeast developmental transition to a filamentous form was clustered. Abstraction of a modular network structure model identified module-organizer proteins and module-connector proteins. The functions of these proteins suggest that they are important for module function and intermodule communication. PMID- 12538876 TI - Parallel changes in gene expression after 20,000 generations of evolution in Escherichiacoli. AB - Twelve populations of Escherichia coli, derived from a common ancestor, evolved in a glucose-limited medium for 20,000 generations. Here we use DNA expression arrays to examine whether gene-expression profiles in two populations evolved in parallel, which would indicate adaptation, and to gain insight into the mechanisms underlying their adaptation. We compared the expression profile of the ancestor to that of clones sampled from both populations after 20,000 generations. The expression of 59 genes had changed significantly in both populations. Remarkably, all 59 were changed in the same direction relative to the ancestor. Many of these genes were members of the cAMP-cAMP receptor protein (CRP) and guanosine tetraphosphate (ppGpp) regulons. Sequencing of several genes controlling the effectors of these regulons found a nonsynonymous mutation in spoT in one population. Moving this mutation into the ancestral background showed that it increased fitness and produced many of the expression changes manifest after 20,000 generations. The same mutation had no effect on fitness when introduced into the other evolved population, indicating that a mutation of similar effect was present already. Our study demonstrates the utility of expression arrays for addressing evolutionary issues including the quantitative measurement of parallel evolution in independent lineages and the identification of beneficial mutations. PMID- 12538877 TI - Essential role for Atox1 in the copper-mediated intracellular trafficking of the Menkes ATPase. AB - The metallochaperone Atox1 directly interacts with the copper-transporting ATPases and plays a critical role in perinatal copper homeostasis. To determine the cell biological mechanisms of Atox1 function, intracellular copper metabolism, and Menkes ATPase abundance, localization and trafficking were examined in immortalized fibroblast cell lines derived from Atox1(+/+) and Atox1( /-) embryos. Consistent with the proposed role for Atox1 in copper delivery to the secretory pathway, a marked increase in intracellular copper content secondary to impaired copper efflux was observed in Atox1-deficient cells. Although the localization of the Menkes ATPase was identical in Atox1(+/+) and Atox1(-/-) cells under conditions of equivalent intracellular copper content, a significant impairment in copper-mediated Menkes ATPase trafficking was observed in the absence of Atox1. When quantitative confocal immunofluorescence was used, significant differences in the time and dose-dependent trafficking of the Menkes ATPase from the Golgi compartment in response to copper were observed between Atox1(+/+) and Atox1(-/-) cells. These data reveal an essential role for Atox1 in establishing the threshold for copper-dependent movement of the copper transporting ATPases within the secretory compartment and that, in the absence of Atox1, this movement alone is not sufficient to restore normal copper efflux. Taken together, these findings provide a cell biological model for the role of this metallochaperone under the physiological conditions of copper limitation in mammalian cells. PMID- 12538878 TI - Endocannabinoid signaling via cannabinoid receptor 1 is involved in ethanol preference and its age-dependent decline in mice. AB - Cannabinoids and ethanol can activate the same reward pathways, which could suggest endocannabinoid involvement in the rewarding effects of ethanol. The high ethanol preference of young (6-10 weeks) C57BL6J mice is reduced by the cannabinoid receptor 1 (CB1) antagonist SR141716A to levels observed in their CB1 knockout littermates or in old (26-48 weeks) wild-type mice, in both of which ethanol preference is unaffected by SR141716A. Similarly, SR141716A inhibits food intake in food-restricted young, but not old, wild-type mice. There are no age dependent differences in the tissue levels of the endocannabinoids anandamide and 2-arachidonoylglycerol or the density of CB1 in the hypothalamus, limbic forebrain, amygdala, and cerebellum. CB1-stimulated guanosine 5'-[gamma thio]triphosphate (GTP[gammaS]) binding is selectively reduced in the limbic forebrain of old compared with young wild-type mice. There is no age-dependent difference in G(i) or G(o) subunit protein expression in the limbic forebrain, and the selective reduction in GTP[gammaS] labeling in tissue from old mice is maintained in a receptorG protein reconstitution assay by using functional bovine brain G protein. These findings suggest that endocannabinoids acting at CB1 contribute to ethanol preference, and decreased coupling of CB1 to G proteins in the limbic forebrain by mechanisms other than altered receptor or G protein levels may be involved in the age-dependent decline in the appetite for both ethanol and food. PMID- 12538879 TI - Components of the Rb pathway are critical targets of UV mutagenesis in a murine melanoma model. AB - Epidemiological studies support a link between melanoma risk and UV exposure early in life, yet the molecular targets of UV's mutagenic actions are not known. By using well characterized murine models of melanoma, we provide genetic and molecular evidence that identifies components of the Rb pathway as the principal targets of UV mutagenesis in murine melanoma development. In a melanoma model driven by H-RAS activation and loss of p19(ARF) function, UV exposure resulted in a marked acceleration in melanoma genesis, with nearly half of these tumors harboring amplification of cyclin-dependent kinase (cdk) 6, whereas none of the melanomas arising in the absence of UV treatment possessed cdk6 amplification. Moreover, UV-induced melanomas showed a strict reciprocal relationship between cdk6 amplification and p16(INK4a) loss, which is consistent with the actions of UV along the Rb pathway. Most significantly, UV exposure had no impact on the kinetics of melanoma driven by H-RAS activation and p16(INK4a) deficiency. Together, these molecular and genetic data identify components of the Rb pathway as critical biological targets of UV-induced mutagenesis in the development of murine melanoma in vivo. PMID- 12538881 TI - Excitation-transcription coupling: signaling by ion channels to the nucleus. AB - Changes in the concentration of intracellular Ca2+ ([Ca2+]i) in response to various stimuli play a role in regulating numerous cellular processes, including the activation of gene expression. In neurons, the extraordinary diversity of the response to Ca2+ signaling depends on the location, intensity, and duration of the Ca2+ transient. Interestingly, Ca2+-dependent gene transcription appears to be sensitive both to increases in nuclear Ca2+, which occur after relatively intense stimuli, and to highly localized increases in Ca2+ near the sites of Ca2+ influx. Activation of intracellular signaling pathways by specific types of Ca2+ channels depends on localization of specific Ca2+ receptors close to the channel mouth. The dual regulation of signaling pathways by Ca2+ near channels and in the nucleus may permit neurons to precisely tailor transcriptional activation to specific types of electrical or chemical stimuli and at the same time ensure that only robust stimuli that generate nuclear Ca2+ elevations are converted into long term changes in gene expression. PMID- 12538880 TI - Deficiency of acyl CoA:cholesterol acyltransferase 2 prevents atherosclerosis in apolipoprotein E-deficient mice. AB - Deficiency of acyl CoA:cholesterol acyltransferase 2 (ACAT2) in mice results in a reduction in cholesterol ester synthesis in the small intestine and liver, which in turn limits intestinal cholesterol absorption, hepatic cholesterol gallstone formation, and the accumulation of cholesterol esters in the plasma lipoproteins. Here we examined the contribution of ACAT2-derived cholesterol esters to atherosclerosis by crossing ACAT2-deficient (ACAT2(-/-)) mice with apolipoprotein (apo) E-deficient (ApoE(-/-)) mice, an atherosclerosis-susceptible strain that has impaired apoE-mediated clearance of apoB-containing lipoproteins. ACAT2(-/-) ApoE(-/-) mice and ACAT2(+/+) ApoE(-/-) (control) mice had similar elevations of plasma apoB and total plasma lipids; however, the lipid cores of the apoB containing lipoproteins in ACAT2(-/-) ApoE(-/-) mice contained primarily triglycerides rather than cholesterol esters. At 30 wk of age, only the control mice had significant atherosclerosis, which was nearly absent in ACAT2(-/-) ApoE( /-) mice. ACAT2 deficiency in the apoE-deficient background also led to a compensatory increase in the activity of lecithincholesterol acyltransferase, the major plasma cholesterol esterification enzyme, which increased high-density lipoprotein cholesterol esters. Our results demonstrate the crucial role of ACAT2 derived cholesterol esters in the development of atherosclerosis in mice and suggest that triglyceride-rich apoB-containing lipoproteins are not as atherogenic as those containing cholesterol esters. Our results also support the rationale of pharmacological inhibition of ACAT2 as a therapy for atherosclerosis. PMID- 12538882 TI - FXYD proteins: new tissue-specific regulators of the ubiquitous Na,K-ATPase. AB - Maintenance of the Na+ and K+ gradients between the intracellular and extracellular milieus of animal cells is a prerequisite for basic cellular homeostasis and for functions of specialized tissues. The Na,K-ATPase, an oligomeric P-type adenosine triphosphatase (ATPase), is composed of a catalytic alpha subunit and a regulatory beta subunit and is the main player that fulfils these tasks. A variety of regulatory mechanisms are necessary to guarantee appropriate Na,K-ATPase expression and activity adapted to changing physiological demands. Recently, a regulatory mechanism was defined that is mediated by interaction of Na,K-ATPase with small proteins of the FXYD family, which possess a single transmembrane domain and so far have been considered as channels or regulators of ion channels. The mammalian FXYD proteins FXYD1 through FXYD7 exhibit tissue-specific distribution. Phospholemman (FXYD1) in heart and skeletal muscle, the gamma subunit of Na,K-ATPase (FXYD2) and corticosteroid hormone induced factor (FXYD4, also known as CHIF) in the kidney, and FXYD7 in the brain associate preferentially with the widely expressed Na,K-ATPase alpha1-beta1 isozyme and modulate its transport activity in a way that conforms to tissue specific requirements. Thus, tissue- and isozyme-specific interaction of Na,K ATPase with FXYD proteins contributes to proper handling of Na+ and K+ by the Na,K-ATPase, and ensures correct function in such processes as renal Na+ reabsorption, muscle contraction, and neuronal excitability. PMID- 12538883 TI - Low free energy cost of very long loop insertions in proteins. AB - Long insertions into a loop of a folded host protein are expected to have destabilizing effects because of the entropic cost associated with loop closure unless the inserted sequence adopts a folded structure with amino- and carboxy termini in close proximity. A loop entropy reduction screen based on this concept was used in an attempt to retrieve folded sequences from random sequence libraries. A library of long random sequences was inserted into a loop of the SH2 domain, displayed on the surface of M13 phage, and the inserted sequences that did not disrupt SH2 function were retrieved by panning using beads coated with a phosphotyrosine containing SH2 peptide ligand. Two sequences of a library of 2 x 10(8) sequences were isolated after multiple rounds of panning, and were found to have recovery levels similar to the wild-type SH2 domain and to be relatively intolerant to further mutation in PCR mutagenesis experiments. Surprisingly, although these inserted sequences exhibited little nonrandom structure, they do not significantly destabilize the host SH2 domain. Additional insertion variants recovered at lower levels in the panning experiments were also found to have a minimal effect on the stability and peptide-binding function of the SH2 domain. The additional level of selection present in the panning experiments is likely to involve in vivo folding and assembly, as there was a rough correlation between recovery levels in the phage-panning experiments and protein solubility. The finding that loop insertions of 60-80 amino acids have minimal effects on SH2 domain stability suggests that the free energy cost of inserting long loops may be considerably less than polymer theory estimates based on the entropic cost of loop closure, and, hence, that loop insertion may have provided an evolutionary route to multidomain protein structures. PMID- 12538884 TI - High-pressure 1H NMR study of pressure-induced structural changes in the heme environments of metcyanomyoglobins. AB - The effect of pressure on the heme environment structure of sperm whale and horse heart metcyanomyoglobins was investigated up to 300 MPa by high-pressure (1)H NMR spectroscopy. Pressure-induced changes in the distances between the observed protons and the heme iron atom were estimated from changes in the dipolar shift due to the paramagnetic effect on the protons. The changes showed that the heme peripheral structure as a whole was compressed by pressure; the movements of the protons in the heme peripheral residues were in the range of +0.16 to -0.54 A/300 MPa. One-dimensional compressibilities for the protons, excluding the protons of the distal His residue, were in the range of 1.0 x 10(-4) to 6.1 x 10(-4)/MPa. The movements of the protons induced by pressure correlated well with the distance between the protons and cavities in the protein. The distal His residue (His 64) moved toward the outside of the heme pocket, but remained in the pocket even at 300 MPa. This movement was driven dominantly by a change in the dihedral angle around the C(alpha)-C(beta) rotational bond of the residue. Comparative work on horse heart metcyanomyoglobin implied that the conformational change of the His 64 imidazole ring was larger in the horse heart metcyanomyoglobin than in the sperm whale metcyanomyoglobin. PMID- 12538885 TI - Expression, solubilization, and biochemical characterization of the tight junction transmembrane protein claudin-4. AB - The tight junction tetraspan protein claudin-4 creates a charge-selective pore in the paracellular pathway across epithelia. The structure of the pore is unknown, but is presumed to result from transcellular adhesive contacts between claudin's extracellular loops. Here we report the expression of claudin-4 by baculovirus infection of Sf9 cells and describe the biochemical analysis suggesting it has a hexameric quaternary configuration. We show the detergent perfluoro-octanoic acid is able to maintain oligomeric claudin species. Sucrose velocity centrifugation and laser light scattering are also used to investigate the oligomeric state of claudin-4. In contrast to proteins of similar topology, such as gap junction family connexins, the oligomeric state of claudins appears more dynamic. These data suggest the structural organization of claudins in tight junction pores is unique. PMID- 12538886 TI - Protein conformational changes studied by diffusion NMR spectroscopy: application to helix-loop-helix calcium binding proteins. AB - Pulsed-field gradient (PFG) diffusion NMR spectroscopy studies were conducted with several helix-loop-helix regulatory Ca(2+)-binding proteins to characterize the conformational changes associated with Ca(2+)-saturation and/or binding targets. The calmodulin (CaM) system was used as a basis for evaluation, with similar hydrodynamic radii (R(h)) obtained for apo- and Ca(2+)-CaM, consistent with previously reported R(h) data. In addition, conformational changes associated with CaM binding to target peptides from myosin light chain kinase (MLCK), phosphodiesterase (PDE), and simian immunodeficiency virus (SIV) were accurately determined compared with small-angle X-ray scattering results. Both sets of data demonstrate the well-established collapse of the extended Ca(2+)-CaM molecule into a globular complex upon peptide binding. The R(h) of CaM complexes with target peptides from CaM-dependent protein kinase I (CaMKI) and an N terminal portion of the SIV peptide (SIV-N), as well as the anticancer drug cisplatin were also determined. The CaMKI complex demonstrates a collapse analogous to that observed for MLCK, PDE, and SIV, while the SIV-N shows only a partial collapse. Interestingly, the covalent CaM-cisplatin complex shows a near complete collapse, not expected from previous studies. The method was extended to related calcium binding proteins to show that the R(h) of calcium and integrin binding protein (CIB), calbrain, and the calcium-binding region from soybean calcium-dependent protein kinase (CDPK) decrease on Ca(2+)-binding to various extents. Heteronuclear NMR spectroscopy suggests that for CIB and calbrain this is likely because of shifting the equilibrium from unfolded to folded conformations, with calbrain forming a dimer structure. These results demonstrate the utility of PFG-diffusion NMR to rapidly and accurately screen for molecular size changes on protein-ligand and protein-protein interactions for this class of proteins. PMID- 12538887 TI - Turn stability in beta-hairpin peptides: Investigation of peptides containing 3:5 type I G1 bulge turns. AB - The turn-forming ability of a series of three-residue sequences was investigated by substituting them into a well-characterized beta-hairpin peptide. The starting scaffold, bhpW, is a disulfide-cyclized 10-residue peptide that folds into a stable beta-hairpin with two antiparallel strands connected by a two-residue reverse turn. Substitution of the central two residues with the three-residue test sequences leads to less stable hairpins, as judged by thiol-disulfide equilibrium measurements. However, analysis of NMR parameters indicated that each molecule retains a significant folded population, and that the type of turn adopted by the three-residue sequence is the same in all cases. The solution structure of a selected peptide with a PDG turn contained an antiparallel beta hairpin with a 3:5 type I + G1 bulge turn. Analysis of the energetic contributions of individual turn residues in the series of peptides indicates that substitution effects have significant context dependence, limiting the predictive power of individual amino acid propensities for turn formation. The most stable and least stable sequences were also substituted into a more stable disulfide-cyclized scaffold and a linear beta-hairpin scaffold. The relative stabilities remained the same, suggesting that experimental measurements in the bhpW context are a useful way to evaluate turn stability for use in protein design projects. Moreover, these scaffolds are capable of displaying a diverse set of turns, which can be exploited for the mimicry of protein loops or for generating libraries of reverse turns. PMID- 12538888 TI - Directing the mode of nitrite binding to a copper-containing nitrite reductase from Alcaligenes faecalis S-6: characterization of an active site isoleucine. AB - Unlike the heme cd(1)-based nitrite reductase enzymes, the molecular mechanism of copper-containing nitrite reductases remains controversial. A key source of controversy is the productive binding mode of nitrite in the active site. To identify and characterize the molecular determinants associated with nitrite binding, we applied a combinatorial mutagenesis approach to generate a small library of six variants at position 257 in nitrite reductase from Alcaligenes faecalis S-6. The activities of these six variants span nearly two orders of magnitude with one variant, I257V, the only observed natural substitution for Ile257, showing greater activity than the native enzyme. High-resolution (> 1.8 A) nitrite-soaked crystal structures of these variants display different modes of nitrite binding that correlate well with the altered activities. These studies identify for the first time that the highly conserved Ile257 in the native enzyme is a key molecular determinant in directing a catalytically competent mode of nitrite binding in the active site. The O-coordinate bidentate binding mode of nitrite observed in native and mutant forms with high activity supports a catalytic model distinct from the heme cd(1) NiRs. (The atomic coordinates for I257V[NO(2)(-)], I257L[NO(2)(-)], I257A[NO(2)(-)], I257T[NO(2)(-)], I257M[NO(2)( )] and I257G[NO(2)(-)] AfNiR have been deposited in the Protein Data Bank [PDB identification codes are listed in Table 2].) PMID- 12538889 TI - Stabilization of a pH-sensitive apoptosis-linked coiled coil through single point mutations. AB - The apoptosis-associated Par-4 protein has been implicated in cancers of the prostate, colon, and kidney, and in Alzheimer's and Huntington's diseases, among other neurodegenerative disorders. Previously, we have shown that a peptide from the Par-4 C-terminus, which is responsible for Par-4 self-association as well as interaction with all currently identified effector molecules, is natively unfolded at neutral pH, but forms a tightly associated coiled coil at acidic pH and low temperature. Here, we have alternately mutated the two acidic residues predicted to participate in repulsive electrostatic interactions at the coiled coil interhelical interface. Analysis of circular dichroism spectra reveals that a dramatic alteration of the folding/unfolding equilibrium of this peptide can be effected through directed-point mutagenesis, confirming that the two acidic residues are indeed key to the pH-dependent folding behavior of the Par-4 coiled coil, and further suggesting that alleviation of charge repulsion through exposure to either a low pH microenvironment or an electrostatically complementary environment may be necessary for efficient folding of the Par-4 C terminus. PMID- 12538890 TI - Motions and structural variability within toxins: implication for their use as scaffolds for protein engineering. AB - Animal toxins are small proteins built on the basis of a few disulfide bonded frameworks. Because of their high variability in sequence and biologic function, these proteins are now used as templates for protein engineering. Here we report the extensive characterization of the structure and dynamics of two toxin folds, the "three-finger" fold and the short alpha/beta scorpion fold found in snake and scorpion venoms, respectively. These two folds have a very different architecture; the short alpha/beta scorpion fold is highly compact, whereas the "three-finger" fold is a beta structure presenting large flexible loops. First, the crystal structure of the snake toxin alpha was solved at 1.8-A resolution. Then, long molecular dynamics simulations (10 ns) in water boxes of the snake toxin alpha and the scorpion charybdotoxin were performed, starting either from the crystal or the solution structure. For both proteins, the crystal structure is stabilized by more hydrogen bonds than the solution structure, and the trajectory starting from the X-ray structure is more stable than the trajectory started from the NMR structure. The trajectories started from the X-ray structure are in agreement with the experimental NMR and X-ray data about the protein dynamics. Both proteins exhibit fast motions with an amplitude correlated to their secondary structure. In contrast, slower motions are essentially only observed in toxin alpha. The regions submitted to rare motions during the simulations are those that exhibit millisecond time-scale motions. Lastly, the structural variations within each fold family are described. The localization and the amplitude of these variations suggest that the regions presenting large-scale motions should be those tolerant to large insertions or deletions. PMID- 12538891 TI - Trapping of peptide-based surrogates in an artificially created channel of cytochrome c peroxidase. AB - As recently described, the deliberate removal of the proposed electron transfer pathway from cytochrome c peroxidase resulted in the formation of an extended ligand-binding channel. The engineered channel formed a template for the removed peptide segment, suggesting that synthetic surrogates might be introduced to replace the native electron transfer pathway. This approach could be united with the recent development of sensitizer-linked substrates to initiate and study electron transfer, allowing access to unresolved issues about redox mechanism of the enzyme. Here, we present the design, synthesis, and screening of a peptide library containing natural and unnatural amino acids to identify the structural determinants for binding this channel mutant. Only one peptide, (benzimidazole propionic acid)-Gly-Ala-Ala, appeared to interact, and gave evidence for both reversible and kinetically trapped binding, suggesting multiple conformations for the channel protein. Notably, this peptide was the most analogous to the removed electron transfer sequence, supporting the use of a cavity-template strategy for design of specific sensitizer-linked substrates as replacements for the native electron transfer pathway. PMID- 12538892 TI - Accurate and automated classification of protein secondary structure with PsiCSI. AB - PsiCSI is a highly accurate and automated method of assigning secondary structure from NMR data, which is a useful intermediate step in the determination of tertiary structures. The method combines information from chemical shifts and protein sequence using three layers of neural networks. Training and testing was performed on a suite of 92 proteins (9437 residues) with known secondary and tertiary structure. Using a stringent cross-validation procedure in which the target and homologous proteins were removed from the databases used for training the neural networks, an average 89% Q3 accuracy (per residue) was observed. This is an increase of 6.2% and 5.5% (representing 36% and 33% fewer errors) over methods that use chemical shifts (CSI) or sequence information (Psipred) alone. In addition, PsiCSI improves upon the translation of chemical shift information to secondary structure (Q3 = 87.4%) and is able to use sequence information as an effective substitute for sparse NMR data (Q3 = 86.9% without (13)C shifts and Q3 = 86.8% with only H(alpha) shifts available). Finally, errors made by PsiCSI almost exclusively involve the interchange of helix or strand with coil and not helix with strand (<2.5 occurrences per 10000 residues). The automation, increased accuracy, absence of gross errors, and robustness with regards to sparse data make PsiCSI ideal for high-throughput applications, and should improve the effectiveness of hybrid NMR/de novo structure determination methods. A Web server is available for users to submit data and have the assignment returned. PMID- 12538893 TI - NMR solution structure of the activation domain of human procarboxypeptidase A2. AB - The activation domain of human procarboxypeptidase A2, ADA2h, is an 81-residue globular domain released during the proteolytic activation of the proenzyme. The role of this and other similar domains as assistants of the correct folding of the enzyme is not fully understood. The folding pathway of ADA2h was characterized previously, and it was also observed that under certain conditions it may convert into amyloid fibrils in vitro. To gain insight into these processes, a detailed description of its three-dimensional structure in aqueous solution is required so that eventual changes could be properly monitored. A complete assignment of the (1)H and (15)N resonances of ADA2h was performed, and the solution structure, as derived from a set of 1688 nonredundant constraints, is very well defined (pairwise backbone RMSD = 0.67 +/- 0.17 A for residues 10 80). The structure is composed of two antiparallel alpha-helices comprising residues 19-32 and 58-69 packed on the same side of a three-stranded beta-sheet spanning residues 10-15, 50-55, and 73-75. The global fold for the isolated human A2 activation domain is very similar to that of porcine carboxypeptidase B, as well as to the structure of the domain in the crystal of the intact human proenzyme. The observed structural differences relative to the intact human proenzyme are located at the interface between the activation domain and the enzyme and can be related with the activation mechanism. The backbone amide proton exchange behavior of ADA2h was also examined. The global free energy of unfolding obtained from exchange data of the most protected amide protons at pH 7.0 and 298K is 4.9 +/- 0.3 kcal.mole(-1), in good agreement with the values determined by thermal or denaturant unfolding studies. PMID- 12538894 TI - Insights into binding cooperativity of MATa1/MATalpha2 from the crystal structure of a MATa1 homeodomain-maltose binding protein chimera. AB - The Yeast MATa1 and MATalpha2 are homeodomain proteins that bind DNA cooperatively to repress transcription of cell type specific genes. The DNA affinity and specificity of MATa1 in the absence of MATalpha2, however, is very low. MATa1 is converted to a higher affinity DNA-binding protein by its interaction with the C-terminal tail of MATalpha2. To understand why MATa1 binds DNA weakly by itself, and how the MATalpha2 tail affects the affinity of MATa1 for DNA, we determined the crystal structure of a maltose-binding protein (MBP) a1 chimera whose DNA binding behavior is similar to MATa1. The overall MATa1 conformation in the MBP-a1 structure, which was determined in the absence of alpha2 and DNA, is similar to that in the a1/alpha2/DNA structure. The sole difference is in the C-terminal portion of the DNA recognition helix of MATa1, which is flexible in the present structure. However, these residues are not in a location likely to be affected by binding of the MATalpha2 tail. The results argue against conformational changes in a1 induced by the tail of MATalpha2, suggesting instead that the MATalpha2 tail energetically couples the DNA binding of MATalpha2 and MATa1. PMID- 12538895 TI - On the evaluation and optimization of protein X-ray structures for pKa calculations. AB - The calculation of the physical properties of a protein from its X-ray structure is of importance in virtually every aspect of modern biology. Although computational algorithms have been developed for calculating everything from the dynamics of a protein to its binding specificity, only limited information is available on the ability of these methods to give accurate results when used with a particular X-ray structure. We examine the ability of a pKa calculation algorithm to predict the proton-donating residue in the catalytic mechanism of hen egg white lysozyme. We examine the correlation between the ability of the pKa calculation method to obtain the correct result and the overall characteristics of 41 X-ray structures such as crystallization conditions, resolution, and the output of structure validation software. We furthermore examine the ability of energy minimizations (EM), molecular dynamics (MD) simulations, and structure perturbation methods to optimize the X-ray structures such that these give correct results with the pKa calculation algorithm. We propose a set of criteria for identifying the proton donor in a catalytic mechanism, and demonstrate that the application of these criteria give highly accurate prediction results when using unmodified X-ray structures. More specifically, we are able to successfully identify the proton donor in 85% of the X-ray structures when excluding structures with crystal contacts near the active site. Neither the use of the overall characteristics of the X-ray structures nor the optimization of the structure by EM, MD, or other methods improves the results of the pKa calculation algorithm. We discuss these results and their implications for the design of structure-based energy calculation algorithms in general. PMID- 12538896 TI - Crystal structure of a trimeric form of dephosphocoenzyme A kinase from Escherichia coli. AB - Coenzyme A (CoA) is an essential cofactor used in a wide variety of biochemical pathways. The final step in the biosynthesis of CoA is catalyzed by dephosphocoenzyme A kinase (DPCK, E.C. 2.7.1.24). Here we report the crystal structure of DPCK from Escherichia coli at 1.8 A resolution. This enzyme forms a tightly packed trimer in its crystal state, in contrast to its observed monomeric structure in solution and to the monomeric, homologous DPCK structure from Haemophilus influenzae. We have confirmed the existence of the trimeric form of the enzyme in solution using gel filtration chromatography measurements. Dephospho-CoA kinase is structurally similar to many nucleoside kinases and other P-loop-containing nucleotide triphosphate hydrolases, despite having negligible sequence similarity to these enzymes. Each monomer consists of five parallel beta strands flanked by alpha-helices, with an ATP-binding site formed by a P-loop motif. Orthologs of the E. coli DPCK sequence exist in a wide range of organisms, including humans. Multiple alignment of orthologous DPCK sequences reveals a set of highly conserved residues in the vicinity of the nucleotide/CoA binding site. PMID- 12538897 TI - Computational design of a water-soluble analog of phospholamban. AB - Membrane proteins and water-soluble proteins share a similar core. This similarity suggests that it should be possible to water-solubilize membrane proteins by mutating only their lipid-exposed residues. We have developed computational tools to design water-soluble variants of helical membrane proteins, using the pentameric phospholamban (PLB) as our test case. To water solublize PLB, the membrane-exposed positions were changed to polar or charged amino acids, while the putative core was left unaltered. We generated water soluble phospholamban (WSPLB), and compared its properties to its predecessor PLB. In aqueous solution, WSPLB mimics all of the reported properties of PLB including oligomerization state, helical structure, and stabilization upon phosphorylation. We also characterized the truncated mutant WSPLB (21-52) comprising only the former transmembrane segment of PLB. This peptide shows a decreased specificity for forming a pentameric oligomerization state. PMID- 12538898 TI - Structural variation and inhibitor binding in polypeptide deformylase from four different bacterial species. AB - Polypeptide deformylase (PDF) catalyzes the deformylation of polypeptide chains in bacteria. It is essential for bacterial cell viability and is a potential antibacterial drug target. Here, we report the crystal structures of polypeptide deformylase from four different species of bacteria: Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, and Escherichia coli. Comparison of these four structures reveals significant overall differences between the two Gram-negative species (E. coli and H. influenzae) and the two Gram-positive species (S. pneumoniae and S. aureus). Despite these differences and low overall sequence identity, the S1' pocket of PDF is well conserved among the four enzymes studied. We also describe the binding of nonpeptidic inhibitor molecules SB 485345, SB-543668, and SB-505684 to both S. pneumoniae and E. coli PDF. Comparison of these structures shows similar binding interactions with both Gram negative and Gram-positive species. Understanding the similarities and subtle differences in active site structure between species will help to design broad spectrum polypeptide deformylase inhibitor molecules. PMID- 12538899 TI - A reinterpretation of the dimerization interface of the N-terminal domains of STATs. AB - The crystal structures of the N-terminal domain (N-domain) and the core region of the STAT family of transcription factors have been determined previously. STATs can form cooperative higher order structures (tetramers or higher oligomers) while bound to DNA. The crystal packing in the STAT4 N-domain crystal structure, determined at 1.5 A resolution, suggests two alternate organizations of the N domain dimer. We now present the results of site directed mutagenesis of residues predicted to be involved at each dimer interface. Our results indicate that the dimer interface suggested earlier as being physiologically relevant is, in fact, unlikely to be so. Given the alternative model for the N-domain dimer, the ability of the N-domain to mediate interactions of two STAT dimers on DNA remains unchanged. PMID- 12538900 TI - Amino acid sequence and homology modeling of obtustatin, a novel non-RGD containing short disintegrin isolated from the venom of Vipera lebetina obtusa. AB - Disintegrins represent a group of cysteine-rich peptides occurring in Crotalidae and Viperidae snake venoms, and are potent antagonists of several integrin receptors. A novel disintegrin, obtustatin, was isolated from the venom of the Vipera lebetina obtusa viper, and represents the first potent and selective inhibitor of the binding of integrin alpha(1)beta(1) to collagen IV. The primary structure of obtustatin contains 41 amino acids and is the shortest disintegrin described to date. Obtustatin shares the pattern of cysteines of other short disintegrins. However, in contrast to known short disintegrins, the integrin binding loop of obtustatin is two residues shorter and does not express the classical RGD sequence. Using synthetic peptides, a KTS motif was identified as the integrin-binding sequence. A three-dimensional model of obtustatin, built by homology-modeling structure calculations using different templates and alignments, strongly indicates that the novel KTS motif may reside at the tip of a flexible loop. PMID- 12538901 TI - Non-animal origin of animal thioredoxin reductases: implications for selenocysteine evolution and evolution of protein function through carboxy terminal extensions. AB - Thioredoxin reductase (TR) and thioredoxin constitute a major cellular redox system present in all organisms. In contrast to a single form of thioredoxin, there are two TR types: One (bacterial type or small TR) is present in bacteria, archaea, plants, and most unicellular eukaryotes, whereas the second (animal or large TR) is only found in animals and typically contains a carboxy-terminal penultimate selenocysteine encoded by TGA. Surprisingly, we detected sequences of large TRs in various unicellular eukaryotes. Moreover, green algae Chlamydomonas reinhardtii had both small and large TRs, with the latter being a selenoprotein, but no examples of horizontal gene transfer from animals to the green algae could be detected. In addition, phylogenetic analyses revealed that large TRs formed a subgroup of lower eukaryotic glutathione reductases (GRs). The data suggest that the large TR evolved in a lower eukaryote capable of selenocysteine insertion rather than in an animal. The enzyme appeared to evolve by a carboxy-terminal extension of GR such that the resulting carboxy-terminal glutathionelike peptide became an intramolecular substrate for GR and a reductant for thioredoxin. Subsequently, small TRs were lost in an organism that gave rise to animals, large TRs were lost in plants and fungi, and selenocysteine/cysteine replacements took place in some large TRs. Our data implicate carboxy-terminal extension of proteins as a general mechanism of evolution of new protein function. PMID- 12538902 TI - Vibrio cholerae cytolysin is composed of an alpha-hemolysin-like core. AB - The enteric pathogen Vibrio cholerae secretes a water-soluble 80-kD cytolysin, Vibrio cholerae cytolysin (VCC) that assembles into pentameric channels following proteolytic activation by exogenous proteases. Until now, VCC has been placed in a unique class of pore-forming toxins, distinct from paradigms such as Staphyloccal alpha-hemolysin. However, as reported here, amino acid sequence analysis and three-dimensional structure modeling indicate that the core component of the VCC toxin is related in sequence and structure to a family of hemolysins from Staphylococcus aureus that include leukocidin F and alpha hemolysin. Furthermore, our analysis has identified the channel-forming region of VCC and a potential lipid head-group binding site, and suggests a conserved mechanism of assembly and lysis. An additional domain in the VCC toxin is related to plant lectins, conferring additional target cell specificity to the toxin. PMID- 12538903 TI - Structural composition of betaI- and betaII-proteins. AB - Circular dichroism spectra of proteins are sensitive to protein secondary structure. The CD spectra of alpha-rich proteins are similar to those of model alpha-helices, but beta-rich proteins exhibit CD spectra that are reminiscent of CD spectra of either model beta-sheets or unordered polypeptides. The existence of these two types of CD spectra for beta-rich proteins form the basis for their classification as betaI- and betaII-proteins. Although the conformation of beta sheets is largely responsible for the CD spectra of betaI-proteins, the source of betaII-protein CD, which resembles that of unordered polypeptides, is not completely understood. The CD spectra of unordered polypeptides are similar to that of the poly(Pro)II helix, and the poly(Pro)II-type (P2) structure forms a significant fraction of the unordered conformation in globular proteins. We have compared the beta-sheet and P2 structure contents in beta-rich proteins to understand the origin of betaII-protein CD. We find that betaII-proteins have a ratio of P2 to beta-sheet content greater than 0.4, whereas for betaI-proteins this ratio is less than 0.4. The beta-sheet content in betaI-proteins is generally higher than that in betaII-proteins. The origin of two classes of CD spectra for beta-rich proteins appears to lie in their relative beta-sheet and P2 structure contents. PMID- 12538904 TI - 19F NMR study of the leucine-specific binding protein of Escherichia coli: mutagenesis and assignment of the 5-fluorotryptophan-labeled residues. AB - The Escherichia coli L-leucine receptor is an aqueous protein and the first component in the distinct transport pathway for hydrophobic amino acids. L leucine binding induces a conformational change, which enables the receptor to dock to the membrane components. To investigate the ligand-induced conformational change and binding properties of this protein, we used (19)F NMR to probe the four tryptophan residues located in the two lobes of the protein. The four tryptophan residues were labeled with 5-fluorotryptophan and assigned by site directed mutagenesis. The (19)F NMR spectra of the partially ligand free proteins show broadened peaks which sharpen when L-leucine is bound, showing that the labeled wild-type protein and mutants are functional. The titration of L phenylalanine into the 5-fluorotryptophan labeled wild-type protein shows the presence of closed and open conformers. Urea-induced denaturation studies support the NMR results that the wild-type protein binds L-phenylalanine in a different manner to L-leucine. Our studies showed that the tryptophan to phenylalanine mutations on structural units linked to the binding pocket produce subtle changes in the environment of Trp18 located directly in the binding cleft. PMID- 12538905 TI - Key interactions in neocarzinostatin, a protein of the immunoglobulin fold family. AB - Neocarzinostatin (NCS) is a seven-stranded beta-sandwich protein, the folding of which is similar to that of the variable domains of immunoglobulins (Ig). The investigation of the backbone dynamics of apo-NCS [Izadi-Pruneyre et al. (2001) Protein Sci., 10, 2228-2240] enabled us to identify the involvement of long side chain residues in maintaining the rigidity of this beta-protein. In the perspective of using this protein for drug targeting, this raises the following question: do these residues also play a key role in the stabilization of the beta sheet? To investigate this problem, various genetically engineered variants were constructed by mutating these residues to amino acids with shorter aliphatic side chains. These substitutions have no effects on the global fold. However, an important destabilization of the protein, higher than that expected for a simple 'large-to-small' substitution of buried hydrophobic residues, is observed for three mutants, V34A, V21A and V95A. Interestingly, the nature of the residues in these positions is highly conserved in the other Ig-like proteins. The absence of an evolutionary relationship between NCS and the other Ig-like proteins strongly suggests that this hydrophobic core is characteristic of the Ig-fold itself. PMID- 12538906 TI - An analysis of protein domain linkers: their classification and role in protein folding. AB - Recent advances in protein engineering have come from creating multi-functional chimeric proteins containing modules from various proteins. These modules are typically joined via an oligopeptide linker, the correct design of which is crucial for the desired function of the chimeric protein. Here we analyse the properties of naturally occurring inter-domain linkers with the aim to design linkers for domain fusion. Two main types of linker were identified; helical and non-helical. Helical linkers are thought to act as rigid spacers separating two domains. Non-helical linkers are rich in prolines, which also leads to structural rigidity and isolation of the linker from the attached domains. This means that both linker types are likely to act as a scaffold to prevent unfavourable interactions between folding domains. Based on these results we have constructed a linker database intended for the rational design of linkers for domain fusion, which can be accessed via the Internet at http://mathbio.nimr.mrc.ac.uk. PMID- 12538907 TI - Prediction of heterodimerization interfaces of G-protein coupled receptors with a new subtractive correlated mutation method. AB - Recent studies employing differential epitope tagging, selective immunoprecipitation of receptor complexes and fluorescence or bioluminescence resonance energy transfer techniques provide direct evidence for heterodimerization between both closely and distantly related members of the G protein coupled receptor (GPCR) family. Since heterodimerization appears to play a role in modulating agonist affinity, efficacy and/or trafficking properties, the molecular models of GPCRs required to understand receptor function must consider these oligomerization hypotheses. To advance knowledge in this field, we present here a computational approach based on correlated mutation analysis and the structural information contained in three-dimensional molecular models of the transmembrane regions of GPCRs built using the rhodopsin crystal structure as a template. The new subtractive correlated mutation method reveals likely heterodimerization interfaces amongst the different alternatives for the positioning of two tightly packed bundles of seven transmembrane domains next to each other in contact heterodimers of GPCRs. Predictions are applied to GPCRs in the class of opioid receptors. However, in the absence of a known structure of any GPCR dimer, the features of the method and predictions are also illustrated and analyzed for a dimeric complex of known structure. PMID- 12538908 TI - Stabilization of human pancreatic ribonuclease through mutation at its N-terminal edge. AB - Enzyme stability can be an important parameter in the design of recombinant toxins because unstable proteins are often degraded before they can reach their cellular target. There is great interest in the design of human pancreatic ribonuclease variants that could be cytotoxic against tumoral cells. To this end, some residues in the protein need to be substituted, but this may result in a loss of stability. Previous papers have reported the production of N- and C terminal human pancreatic ribonuclease variants with increased thermal stability. Here, we investigated the contribution of the different amino acid changes at the N-terminus of the protein to its thermostability increase. We show that this increase correlates with the helical propensity of the first alpha-helix of the protein. On the other hand, deletion of the four last residues of the protein does not affect its thermal stability. These results set the basis for the design of a human pancreatic ribonuclease template on which amino acid substitutions can be made that could render the enzyme cytotoxic, without an important loss in its stability. PMID- 12538909 TI - Crystine: fibrous biomolecular material from protein crystals cross-linked in a specific geometry. AB - Cysteine substitutions were engineered on the surface of maltose binding protein to produce crystine fibers, linear polymers of folded protein formed within a crystal. Disulfide bond formation between adjacent protein molecules within the lattice was monitored by X-ray crystallography. The cross-linked crystals were resistant to dissolution in water or neutral buffer solutions, even though the cross-linking was one-dimensional. However, crystine fibers were observed by transmission electron microscopy to dissociate from the crystals in acidic solutions. Some fibers remained associated as two-dimensional bundles or sheets, with a repeat unit along the fibers consistent with the packing of the individual protein molecules in the crystal. Neutralization of the acidic solutions caused the fibers to re-associate as a solid. Crystine threads were drawn out of this solution. In scanning electron microscopy images, many individual fibers could be seen unwinding from the ends of some threads. Crystine fibers are a new type of biomolecular material with potential applications wherever the use of proteins in a fibrous form is desirable, for example, the incorporation of enzymes into cloth or filtration material. PMID- 12538910 TI - Mutation of herpesvirus thymidine kinase to generate ganciclovir-specific kinases for use in cancer gene therapies. AB - Understanding the functional and mechanistic properties of the multi-substrate herpes simplex virus type-1 thymidine kinase (HSV-1 TK) remains critical to defining its role as a major pharmacological target in herpesvirus and gene therapies for cancer. An inherent limitation of the activity of HSV-TK is the >70 fold difference in the K(m)s for phosphorylation of thymidine over the pro-drug ganciclovir (GCV). To engineer an HSV-1 TK isoform that is specific for GCV as the preferred substrate, 16 site-specific mutants were generated. The mutations were concentrated at conserved residues involved in nucleoside base binding, Gln125 and near sites 3 and 4 involved in catalysis and substrate binding. The substrate preferences of each mutant enzyme were compared with wild-type HSV-1 TK. One mutant, termed Q7530 TK, had a lower K(m) for GCV than thymidine. Expression of the Q7530 TK in tumor cells indicated comparable metabolism to and improved sensitivity to GCV over wild-type HSV-1 TK, with minimal thymidine phosphorylation activity. A molecular modeling simulation of the different HSV-1 TK active-sites was done for GCV and thymidine binding. It was concluded that mutations at Gln125 and near site 4, especially at Ala168, were responsible for loss of deoxypyrimidine substrate binding. PMID- 12538911 TI - Modification of biologically active peptides: production of a novel lipohexapeptide after engineering of Bacillus subtilis surfactin synthetase. AB - The Bacillus subtilis strain ATCC 21332 produces the lipoheptapeptide surfactin, a highly potent biosurfactant synthesized by a large multimodular peptide synthetase. We report the genetic engineering of the surfactin biosynthesis resulting in the production of a novel lipohexapeptide with altered antimicrobial activities. A combination of in vitro and in vivo recombination approaches was used to construct a modified peptide synthetase by eliminating a large internal region of the enzyme containing a complete amino acid incorporating module. The remaining modules adjacent to the deletion were recombined at different highly conserved sequence motifs characteristic of amino acid incorporating modules of peptide synthetases. The primary goal of this work was to identify permissive fusion sites suitable for the engineering of peptide synthetase genes by genetic recombination. Analysis of the rearranged enzymes after purification from B. subtilis and from the heterologous host Escherichia coli revealed that the selection of the recombination site is of crucial importance for a successful engineering. Only the recombination at a specific HHII x DGVS sequence motif resulted in an active peptide synthetase. The expected lipohexapeptide was produced in vivo and first evidence of a reduced toxicity against erythrocytes and an enhanced lysis of Bacillus licheniformis cells was shown. PMID- 12538912 TI - N-terminal domain of eotaxin-3 is important for activation of CC chemokine receptor 3. AB - Eotaxin-3 belongs to the CC chemokine family, and specifically recognizes CC chemokine receptor (CCR) 3 that is expressed on eosinophils, basophils and helper T type 2 cells. The three-dimensional structure of eotaxin-3 determined by nuclear magnetic resonance has revealed that the N-terminal nine residues preceding the first cysteine comprise an unstructured domain, which is also observed in other chemokine molecules. In order to determine the function of the N-terminal domain of eotaxin-3, we constructed various N-terminal-deletion mutants, and then examined their binding and chemotactic activities toward eosinophils in vitro. Competitive binding studies showed that the binding affinity of truncated mutant toward CCR3 was almost the same as that of wild-type eotaxin-3 even though the N-terminal truncation involved the first through to the ninth residues. In contrast, the chemotactic activity gradually decreased with extension of the N-terminal deletion, and when the deletion extended to the eighth residue, the activity was not detected at all. Thus, the N-terminal nine residues are not critical for binding but the N-terminal eight residues are essential for activation of CCR3. The truncated eotaxin-3 proteins lacking the N terminal eight or nine residues inhibited the chemotactic activity of chemokines that recognize CCR3. The truncated mutants can possibly be used for anti-allergic and anti-HIV-1 therapy. PMID- 12538914 TI - Alpha-helically constrained phage display library. AB - The library described here is a collection of phages with six degenerate codons in gene VIII, specifying amino acids 12, 13, 15-17 and 19 of the major coat protein. The randomized positions are surface exposed in the wild-type protein and thus might be expected to tolerate a great diversity of side chains without compromising phage viability. In agreement with this supposition, the new library showed great diversity of amino acids at the randomized positions and diversity did not diminish noticeably during repeated subculture. Despite their diversity, however, the randomized positions should be strongly constrained conformationally because they lie in an extended alpha-helical portion of the protein, stabilized by numerous inter- and intra-subunit contacts--a presupposition corroborated by circular dichroism spectroscopy of many library members. To reflect this conformational homogeneity and the fact that random amino acids subtend a major fraction of the surface 'landscape' of the particle, we call the new construct an alpha landscape library. It can be used as a source of alpha-helical ligands and substitute antibodies. PMID- 12538913 TI - An engineered IN-1 F(ab) fragment with improved affinity for the Nogo-A axonal growth inhibitor permits immunochemical detection and shows enhanced neutralizing activity. AB - The myelin axonal growth inhibitor NI-220/250 (Nogo-A) has attracted considerable attention in elucidating the mechanisms that account for the lack of plasticity in the adult central nervous system. The cognate monoclonal antibody IN-1, which was obtained prior to the molecular characterization of its Nogo-A antigen, has played a crucial role in this respect. However, this murine IgM/kappa antibody does not only provide an inappropriate format for in vivo studies, its low antigen affinity has also hampered the thorough structure-function analysis of its neutralizing effect toward the Nogo-A inhibitor on a molecular basis. We describe here the affinity maturation of a bacterially produced functional IN-1 F(ab) fragment via protein engineering. A soluble fragment of Nogo-A derived from the central exon 3 of its gene, which was prepared by secretion into the periplasm of Escherichia coli, served as a target in these experiments. After repeated cycles of site-directed random mutagenesis and screening, the mutant II.1.8 of the IN-1 F(ab) fragment was obtained, carrying five side chain substitutions within CDR-L3. Its dissociation constant for the complex with the recombinant Nogo-A fragment was determined in surface plasmon resonance measurements as approximately 1 microM. The affinity of the unmutated IN-1 F(ab) fragment was 8-fold lower. The engineered F(ab) fragment appeared to be well suited for the specific detection of Nogo-A in immunochemical assays and for the histochemical staining of myelin-rich tissue sections. Most importantly, its concentration-dependent neutralizing effect on the Nogo-A inhibitory activity was significantly enhanced in cell culture. This study confirms Nogo-A to be the antigen of the IN-1 antibody and it demonstrates increased potential of the engineered F(ab) fragment as a reagent for promoting axonal regeneration in vivo. PMID- 12538915 TI - [Voluntary consent: basic premise, ideal or utopia?]. PMID- 12538916 TI - [The C0 line: from Delaire's analysis of craniofacial structure to modeling it on a computer spreadsheet]. AB - BACKGROUND: We developed a computer spreadsheet allowing mathematical modelization of Delaire's craniofacial architecture analysis. METHOD: An orthonormed drawing field was defined from the classical C1 line and the new C0 line. The cartesian coordinates defined anatomic points in this orthonormed field. Lines were drawn from the coordinate equations defined on the basis of the laws of plane geometry. These formulae were then entered into the spreadsheet. RESULTS: The spreadsheet automatically calculates the line equations, the angles separating these lines, and the coordinates of the constructed points, in the defined orthonormed field. This calculation is achieved using the coordinates of the user-measured anatomic points. A new order in line drawing allows simplification of manual drawing. DISCUSSION: We present the advantages and inconveniences of this new method which does not modify the final drawing result nor the interpretation of Delaire's analysis. PMID- 12538917 TI - [Temporomandibular ankylosis. A treatment method using the interposition of a Dacron device]. AB - BACKGROUND: Temporomandibular ankylosis with its multiple anatomo-clinical forms is a relatively rare disease. Its major morphopathological, therapeutical and psychological implications rank it among severe illnesses. Its treatment is exclusively surgical. The major therapeutical indication in ankylosis of type I and II Topazian is the neoarticular modelling osteotomy with interposition. MATERIAL AND METHODS: As a material for interposition, over the last 7 years, we have used in 15 patients with 18 ankylosis, concave rectangular Dacron fragments adequately shaped after being taken from a vascular prothesis. RESULTS: The qualities of this material are confirmed by the obtained results: quick resuming of the normal mobility of the mandible, lack of postoperative complications and recurrences. The material is cheap and easy to be obtained. The technique to be used is simple. DISCUSSION: The Dacron texture is soft, elastic in all respects, thick enough, resistant, with long lasting elasticity and integrity, physically and chemically sTable, well tolerated by the body and without foreign body rejection. It is easy to be cut, shaped, modelled and adapted to the bone stump. It is sterilized by autoclaving. It is also well integrated into the host tissue being penetrated by the connective tissue which fastens it to the surface of the neocondyle preventing a relapse. It plays the role of a joint cartilage. PMID- 12538918 TI - [Our experience with geniopexy in the treatment of OSAS]. AB - OBJECTIVE: We examine geniopexy efficacy in OSAS treatment. STUDY DESIGN: Seven patients, aged 36 to 61 years, were studied. The mean body mass index was normal. RESULTS: Surgical late complications (modified symphysis proprioception) occurred in 4 patients. No patient was improved in sleep problems. Retromaxilly and retromandibuly were found in all patients by cephalometric study. DISCUSSION: Our results are quite different from these described successfully by other centers. Geniopexy do not move maxillar and mandibular skeletal bones, which are retruded in our serie. This explains maybe our bad results. Actually, we propose for this patients maxillo-mandibular advancement. PMID- 12538920 TI - [Antibiotic prescription in odontology and stomatology: recommendations and indications]. PMID- 12538921 TI - [Facial mucormycosis: observations from three cases]. AB - The mucormycosis is a rare opportunistic invasive infection caused by fungi of the order Mucorales and characterized by vascular invasion and tissue necrosis. It affects generally the subjects with altered natural resistances, particularly the diabetics patients. The cerebro-rhino-orbital region is the most common site. The clinical signs depend on the intra-tissular and intra-vascular evolution of the fungi. The diagnosis of this disease is asserted by the mycological and anatomo-pathological exams. The treatment is based on the antifungic and the surgical excision of necrotic tissues. We report three observations: one man (42 years) and two women (59 and 60 years). Diabetes was found in two cases. The diagnosis was in every case anatomo-pathologic. Our objective was to study the epidemiological and clinico-pathologic aspects of this serious affection and to discuss its prognosis. PMID- 12538922 TI - [Anterolateral thigh free flap. Surgical technique]. AB - The anterolateral thigh free flap is a cutaneous or fasciocutaneous flap vascularised by one or several perforating arteries arising from the descending branch of the lateral circonflex femoral artery. Venous drainage occurs via the perforators or a similar route to the deep femoral system or the femoral vein. This flap is commonly used in Asia (China, Japan) where for certain teams, it replaces the classical radial forearm flap or the rectus abdominis myocutaneous flap for the reconstruction of head and neck defects after tumor ablation. We briefly describe the anatomy and vascularization of this flap and present the harvesting technique as well as the properties of the flap. PMID- 12538923 TI - [Cysts from the first branchial cleft. Apropos of a case ]. AB - We describe a case of branchial cyst from the first branchial cleft in an 8-year old patient, emphasizing the diagnostic difficulties of this type of lesion and the clinical aspects as well as the details of surgical treatment. The embryogenic origin and the anatomic relation with the facial nerve is recalled. Surgical treatment is indicated. PMID- 12538924 TI - [Thoracic Oncology. 13-15 June 2002, Amsterdam, The Netherlands. Proceedings of a conference]. PMID- 12538925 TI - [New data presented at ASCO 2002 concerning non-small cell bronchial cancers]. PMID- 12538926 TI - [New data presented at ASCO 2002 concerning small cell bronchial cancer]. PMID- 12538927 TI - [New therapeutic options for mesothelioma]. PMID- 12538928 TI - [Screening for bronchial-pulmonary cancer]. PMID- 12538929 TI - [Endobronchial detection and treatment of preneoplastic lesions]. PMID- 12538930 TI - [Netherlands' experience in positron emission tomography in thoracic oncology]. PMID- 12538932 TI - [Adjuvant chemotherapy in the early stages of non-small cell bronchial cancers]. PMID- 12538931 TI - [Principal developments in thoracic surgery]. PMID- 12538933 TI - [Adjuvant or neoadjuvant chemotherapy in non-small cell bronchial cancers? Arguments in favor of choosing neoadjuvant chemotherapy]. PMID- 12538934 TI - [Radio-chemotherapy combination in locally advanced, inoperable, non-small cell bronchial cancers]. PMID- 12538935 TI - [Therapeutic strategies in advanced non-small cell bronchial cancers: platin based combinations]. PMID- 12538936 TI - [Therapeutic strategies in advanced non-small cell bronchial cancers: combinations without platin drugs]. PMID- 12538937 TI - [Chemoprevention in non-small cell bronchial cancers]. PMID- 12538938 TI - [The "caged brain" ]. PMID- 12538939 TI - [MRI characterization of atherosclerotic arteries: diagnosis of plaque rupture]. AB - Plaque stability and its counterpart, vulnerability, depend on the relative amount and morphology of its principal components: lipid core and fibrous cap. Lipids are mainly made of cholesterol esters and cholesterol monohydrates, but phospholipids and triglycerides are also present. Major chemokines, cytokines, enzymes and markers of inflammation are locally produced, which interact and degrade the arterial matrix, but also stimulate smooth muscle cells and osteopontine production. Due to these differences in composition, the lipid cores have various chemical and biophysical properties (with solid, liquid or liquid crystalline phases), and so has the collagenous cap, which influence imaging parameters. MRI has improved its capacity to characterize arterial tissue. This technique can now discriminate the different components of normal and pathological arterial walls through various endogenous or exogenous contrast: intima, media, adventitia, perivascular fat, lipid core, collagenous cap and calcifications of atheromatous plaques, but also precise the elements of plaque resistance to radial stress, or the various factors of inflammation. In the next years, improvement of resolution, new contrast sequences (diffusion or magnetization transfer) or spectroscopy, will compete to allow for the best discrimination of these usual suspects: "the unstable plaques". PMID- 12538940 TI - [Cerebral venous thrombosis]. AB - Cerebral vein and dural sinus thromboses (CVT) appear to be rare, but their identification has increased in recent year, due to the increased use of MRI and to a better knowledge of the variable clinical presentation of CVT.CVT can be diagnosed by MR and MR-angiography. Catheter angiography is reserved for cases whose diagnosis remains uncertain with MR. The individual risk for CVT is determined by genetic factors, but may be increased by numerous predisposing conditions and precipitating factors. In recent CVT series death rate is below 10% and about 80% of patients make a complete recovery. The treatment of CVT includes etiological and symptomatic treatment, antithrombotic therapy (anticoagulants, local thrombolysis) and the treatment of intracranial hypertension. There are few randomized trials upon which to base treatment recommendations. PMID- 12538941 TI - [Epidemiology and pathophysiology of spontaneous cervical artery dissection]. AB - Spontaneous cervical artery dissections are produced by the penetration of circulating blood into the vessel wall of one or more cervical arteries, without a preceding major trauma. Dissection is one of the most frequent etiologies of ischemic stroke in young patients. Its annual incidence is about 3 per 100,000. Pathophysiology of cervical artery dissection remains misunderstood. Triggering factors such as minor trauma and infection have been identified. However, they are too trivial to explain alone the occurrence of a mural hematoma. Several abnormalities suggesting an underlying arteriopathy related to an extracellular matrix defect which could predispose to dissection have been reported: arterial redundancies, intracranial aneurysms, aortic root dilatation, common carotid artery distensibility increase, fibromuscular dysplasia, inherited connective tissue disorders, ultrastructural dermal connective tissue abnormalities. Other factors associated with dissection, such as migraine, hyperhomocysteinemia and alpha-1 antitrypsin deficiency, suggest arterial wall fragility secondary to hyperactivity of some proteases. If an underlying arteriopathy is likely, its nature remains unidentified to date and does not seem to be unique. PMID- 12538942 TI - [Natural history of cervical arterial dissections. Review of the literature and preliminary results from a national study group]. AB - Few studies have been devoted to the natural history of cervical artery dissection (CAD). This review shows that apart from neurological damage due to initial stroke, the prognosis of DAC is good. Indeed, the risk of recurrent DAC or recurrent stroke is very low, under 1% per year in patients without typical connective tissue disease. Patients with connective tissue disease (Ehlers Danlos syndrome or Marfan syndrome) or familial history of arterial dissection have an increased risk of recurrent CAD. However, the number of cases due to these diseases remains very low. The other risk factors for recurrent CAD or stroke remain unknown. The benefit of antithrombotic treatments at the acute phase of CAD or for secondary prevention has been never assessed in a randomized clinical trial. PMID- 12538943 TI - [Imaging of cervical arterial dissections: multi-center study and review of the literature]. AB - Multiple imaging techniques are available to diagnose cervical arterial dissections. It may be possible to image the vessel wall hematoma (US, MRI, 2D or 3D MRA source images, CTA source images), the narrowed arterial lumen (MRA, CTA, catheter angiography), or the hemodynamic impact of the affected vessel. There is no available guideline in the literature regarding the imaging work-up at diagnosis or follow-up. The Societe Francaise Neurovasculaire and the Societe Francaise de Neuroradiologie have sponsored a multi-center study to evaluate current imaging strategies. Results from several centers, while being variable, do share some similarities: the preferred use of non-invasive or minimally invasive techniques to evaluate the vessel wall and the vascular lumen and/or hemodynamic impact, the use of fat suppressed sequences at MRI, the use of Gadolinium enhanced MRA, the limited interest for CT techniques, and the still important use of catheter angiography in selected cases. PMID- 12538944 TI - [Medical management of cervical arterial dissections]. AB - Medical management of cervical arterial dissections is not standardized and has not been the subject of randomized trials. Management is mainly based on the presumed pathophysiology of secondary cerebral infarcts associated with dissections and the individual experience of each treating team. First, a review of the literature regarding medical management of acute and chronic dissections is presented. Then, results from a national study sponsored by the Societe Francaise Neuro-Vasculaire and the Societe Francaise de Neuro-Radiologie evaluating the medical management of this pathology in French neuro-vascular centers will be presented. These data will be useful to generate practical management recommendations and establish guidelines for further studies. PMID- 12538945 TI - Interobserver agreement in the assessment of lobar versus deep location of intracerebral haematomas on CT. AB - In patients with supratentorial intracerebral haemorrhage (ICH), it is important to discriminate superficial (lobar) and deep (basal ganglia) location, since this has consequences for research and prognosis. Haemorrhages at these sites have different causes and different risk factors. We studied the interobserver variation between three radiologists in classifying fifty large haematomas on CT as deep or lobar. The kappa values were almost perfect, ranging from 0.88 to 0.96. We conclude that the assessment of CT by radiologist is a reliable method to discriminate between lobar versus deep origin even for large intracerebral haematomas. PMID- 12538946 TI - [Natural evolution of an aneurysm of the V4 segment of the vertebral artery into a giant aneurysm. A case report]. AB - We report the case of a 78-year-old female with an aneurysm of the V4 segment of the vertebral artery and a six-year follow-up without any surgical or endovascular treatment. The natural evolution of a small aneurysm into a giant aneurysm is rarely observed. We discuss the pathophysiology of the development and growth of such aneurysms through a review of the literature. PMID- 12538947 TI - [Traumatic false aneurysms of intracranial artery]. AB - The authors report a case of intracranial traumatic aneurysm at young a 21 years old patient. He presented with recurrent epistaxis 3 months after craniofacial trauma. An aneurysm of the left carotid artery was diagnosed at cerebral angiography. Endovascular treatment was performed with favorable outcome. In spite of their rarity, posttraumatic intracranial aneurysms should be suspected in patients with recent or remote history of trauma, even minor, presenting with secondary clinical deterioration, in order to carry out emergent angiographic exploration, before contemplating definitive surgical or endovascular treatment. PMID- 12538948 TI - MR spectroscopic pulvinar sign in a case of variant Creutzfeldt-Jakob disease. AB - We report MR spectroscopic findings in a patient hospitalized with biopsy-proven variant Creutzfeldt-Jakob (vCJD) disease. N-acetyl aspartate was markedly decreased in the postero-medial part of the thalami (pulvinar) but was not diminished in the parieto-occipital white matter and cortical grey matter. These observations, which are in accordance with the pathological findings in this disease, suggest that MR spectroscopy, a highly sensitive method for the detection of subtle brain metabolic dysfunction, could be of interest for the diagnosis, prognosis and therapeutic follow-up of vCJD. PMID- 12538950 TI - [Cystic pancreatic tumors]. AB - Cystic tumors of the Pancreas (CTP) are rare (less than 5% of all pancreatic tumors). We have limited our study to CTP of epithelial origin which represent 90% of all CTP. These can be divided into three subgroups: (1) Benign tumors with no risk of malignant progression (serous cystadenoma). (2) Tumors with risk of malignant degeneration (mucinous cystadenoma, intraductal papillary mucinous tumors (IPMT), and pancreatic solid-cystic papillary tumor. (3) Malignant tumors (cystadenocarcinoma, IPMT with malignant degeneration). The latter two groups of CTP require radical resection while serous cystadenoma does not require surgical intervention unless symptomatic. The ability to determine preoperatively the exact nature of a CTP is of tremendous importance; cytologic examination and biochemical assays of cyst fluid aid greatly in this determination. Better understanding of the biologic evolution of CTP has increased the indications for surgical resection but the risks of postoperative morbidity and mortality in these patients must not be underestimated. PMID- 12538951 TI - [What is the best timing to perform laparoscopic cholecystectomy in acute cholecystitis?]. AB - Early laparascopic cholecystectomy within five days of onset of symptoms of acute cholecystitis has proved superior to open cholecystectomy. Feasability is well established; the conversion rate to open cholecystectomy and the post-operative morbidity are no higher than with delayed laparascopy after initial medical management. Total hospitalization is, on the other hand, significantly shorter. Only one study showed an increase in operative time with early surgery (on average 15 minutes). Both preoperative and perioperative risk factors for conversion to laparotomy were identified. Prompt laparascopic intervention avoids the complications which may arise with initial medical management, i.e., failure of medical management or biliary peritonitis. Early laparascopic cholecystectomy should be considered the gold standard for management of acute cholecystitis. PMID- 12538952 TI - [Epiploic appendagitis: a radiologic diagnosis]. AB - The incidence of primary epiploic appendagitis is underestimated. The clinical and biological presentation of epiploic appendagitis is non-specific and may mimic acute appendicitis or diverticulitis. Imaging with ultrasound or computed tomography may suggest the diagnosis and prevent unnecessary surgery; medical management of symptoms is usually sufficient. PMID- 12538953 TI - [55 year-old male presenting with an abdominal mass]. PMID- 12538954 TI - [Rectus abdominis myocutaneous flap reconstruction after pelvic surgery for cancer]. PMID- 12538955 TI - [Organ harvesting: (1) liver]. PMID- 12538956 TI - [The lateral attachments of the so-called "Mesorectum"]. PMID- 12538957 TI - [Obstruction due to interssuception of a colonic lipoma]. PMID- 12538959 TI - [104th French Congress of Surgery]. PMID- 12538960 TI - [2002 Congress of the American College of Surgeons]. PMID- 12538961 TI - [Favorable course after surgical treatment of trocar site metastasis, first sign of non-suspected gallbladder cancer at the time of laparoscopic cholecystectomy]. PMID- 12538962 TI - [SIAARTI recommendations for postoperative pain management]. PMID- 12538963 TI - Training in BLS, ALS and the importance of early defibrillation. PMID- 12538964 TI - Informing patients about risks and complications of anaesthesia. AB - BACKGROUND: This study examines the quality of patients' informed consent in patients undergoing minor surgical procedure. Information provided on risks and complications inherent in anaesthesia-care and patients' awareness of specific risks involved in anaesthesia were recorded, and the influence of a questionnaire form was evaluated. METHODS: This is a prospective controlled study. In the first part (Group I: n=71) social and demographic data, complications and risks communicated to the patient in the preanaesthetic evaluation, as well as patients' satisfaction with the information obtained, were noted. In the second part of the study (Group II: n=201) patients were also asked to answer a questionnaire form concerning their own state of health and their knowledge of risks in anaesthesia. The statistical analysis was conducted using Pearson's Chi square test or t test, and by fitting suitable logistic models. RESULTS: The frequency of the communication of at least one of the risks and complications of anaesthesia has been 43.7% in GI and 59,7% in GII, p=0.019. Patients in class ASA III were informed more (77%, c.i. 59-90) as compared to those in class ASA I-II (52%, c.i. 45-59) (p=0.008); 37% in class ASA I-II and 65% in class ASA III desire the presence of one of their relatives during the preassessment (p=0.016). CONCLUSIONS: Our results indicate that most patients are glad to be informed, even if they are unaware of the risks and complications of anaesthesia. The questionnaire form seems to be an important tool for patient education. Ethical questions are proposed. PMID- 12538966 TI - Ambulatory surgery with long acting regional anesthesia. AB - Ambulatory surgery has grown dramatically in the past 3 decades; however, advances in postoperative pain treatment have not kept pace with the proliferation of outpatient procedures. Two techniques that may offer a solution to part of this problem are long acting peripheral nerve blocks (PNB) and outpatient continuous peripheral nerve blocks (CPNB), but the safety of sending patients home with blocked extremities has also remained controversial. Unfortunately, only a few large, prospective studies have examined this issue directly. Those that have addressed this particular question support the concept that regional anesthesia and discharge with an insensate limb may be done safely. Our group prospectively studied 2,382 long-acting PNB with ropivacaine in both the upper and lower extremity noting a low incidence of block failure, rare use of opioids in the recovery unit and high patient satisfaction. This study also demonstrated a low incidence of accidental injury to the blocked extremity and a rare block complication rate after discharge (0.2%). Only 1 patient in this data set fell while exiting a car. Patients in our study appeared to uniformly protect themselves from further injury despite having a blocked extremity. In conclusion, PNB and perineural catheter techniques are an exciting aspect of ambulatory anesthesia and acute pain management that has undergone rapid development in recent years. Successful application of these techniques will require a substantial educational investment by anesthesiologists and anesthesiologists in training. The rewards in reduced postoperative pain, improved patient satisfaction, and anesthesiologist professional development make this endeavor worthy of our attention. PMID- 12538965 TI - Transesophageal atrial pacing in the management of re-entry supraventricular tachyarrhythmias occurring during general anesthesia. AB - BACKGROUND: Supraventricular tachyarrhythmias (SVTs) represent an intraoperative risk factor that should be always prevented/managed. The commonly used anti arrhythmic drugs are accompanied by intrinsic hazards, such as pro-arrhythmic and toxic effects or unpredictable onset and duration of action. We underline the therapeutic use of transesophageal atrial pacing (TAP) for the interruption of particular re-entry SVTs occurred during surgical procedures in general anaesthesia. METHODS: Our study was carried out in 25 patients characterized by a personal clinical history of transient tachyarrhythmic episodes, subjected to general anaesthesia obtained by midazolam, propofol, N2O e O2, sevoflurane, fentanil and vecuronium bromide. We used TAP bursts of 3-5 sec, their minimal pacing rate being equivalent to the tachyarrhythmia cycle length, with an impulse intensity ranging from 18 to 25 mA. In such conditions, the re-entry was interrupted by the induction of refractoriness of the wave-front that sustained the underlying arrhythmogenic circuit. RESULTS: During the study, the following arrhythmias occurred in 7 out of all patients: 1 type I atrial flutter, 3 nodal tachycardias, 1 antidromic and 2 orthodromic atrioventricular tachycardias, respectively. TAP assured either atrial capture or prompt suppression of arrhythmias in all cases. Low intensity impulses did not ever allow ventricular capture. CONCLUSIONS: TAP can be considered as a valid therapeutic device for the management of re-entry SVTs occurred during general anaesthesia, resulting it effective, safe and easy-practicable. PMID- 12538967 TI - Eight months of emergency services by ambulance (with doctor on board) of the Emergency Department of Prato, Italy. AB - BACKGROUND: The aim of the present study was to analyse the types of calls attended by the authors in their ambulance, assigned to one sector of the urban area of Prato. METHODS: We analysed 1060 consecutive calls carried out by our ambulance; 33 calls (3.1%), cancelled for various reasons, were excluded from statistical analysis. The calls regarded 1027 patients, 549 (53.5%) males and 478 (46.5%) females. RESULTS: 46% of calls concerned non trauma cases, 17% trauma cases and 7.2% transfers between hospitals. Calls in the non trauma group were for dyspnea (16.7%), unconsciousness (16.6%), chest pain (11.8%), cerebrovascular pathology (7.7%), mental disorders (7.1%), abdominal pain (5.1%), use of psychotropic substances (4%) and convulsions (3.2%). Support to patients with terminal cancer accounted for 2.3% of cases, metabolic disorders 2.2%, hypertensive events 2.1%, tachycardia 1.9%, vertigo 1.9%, allergies 1%, obstetric pathology 0.5% and cardiorespiratory resuscitation 2,2%. Thirteen point seven percent were miscellaneous, including poisonings, migraines, haemorrhages and flu syndromes. Trauma cases included road accidents (62.6%), falls (27%), aggression (6.9%), work-related accidents (3.5%). The percentage of trauma cases (17%) was similar to that reported for a Swedish urban area (20%). Cases of cardiorespiratory resuscitation were 1.65% of our calls, compared to 1.73% and 1.8% reported in Northeastern Germany and Taiwan. CONCLUSIONS: Although prehospital emergency medical services are organised differently in different countries, our data suggest that emergency medical services in our area have a percentage distribution of case types similar to services in other countries. PMID- 12538968 TI - Role of desmopressin in the treatment of a case of septic shock refractory to catecholamines. AB - The authors describe a case of septic shock refractory to high-dose catecholamines after adequate volemic filling, rapidly corrected with the administration of desmopressin at a dose of 0.02 U/min. There are, in the literature, several cases of septic shock refractory to high-dose catecholamines successfully treated with arginine-vasopressin, a selective V1 agonist. This agent can potentially induce coronary vasoconstriction and presents a very difficult weaning, that imposes a slow reduction of the infusion amount. On the contrary, desmopressin does not present those adverse effects and is able to maintain a high renal blood flow, for its prevalent action on V2 instead of V1 receptors. PMID- 12538969 TI - The stiff-person syndrome. Case report. AB - The stiff-person syndrome is a rare and disabling disorder, characterized by muscle rigidity with superimposed painful spasms involving axial and limb musculature. The clinical symptoms are continuous contraction of agonist and antagonist muscles caused by involuntary motor-unit firing at rest and the spasms that are precipitated by tactile stimuli, passive strach, volitional movement of affected or unaffected muscles, startling noises and emotional stimuli. Both the rigidity and the spasms are relieved by sleep, general anaesthesia, myoneural blockade and peripheral nerve blockade. The cause of the stiff-person syndrome is unknown but an autoimmune pathogenesis is suspected because 1) the presence in the cerebrospinal fluid (CSF) of antibodies against glutamic acid decarboxylase (GAD), the rate-limiting enzyme for the synthesis of the inhibitory neurotrasmitter gamma-aminobutyric acid (GABA), 2) the association of the disease with other autoimmune disorders, 3) the presence of various autoantibodies and 4) a strong immunogenetic association. The stiff-person syndrome is clinically elusive but potentially treatable and should be considered in patients with unexplained stiffness and spasms. Drugs that enhance GABA neurotransmission, such as diazepam, vigabatrin and baclofen, provide modest relief of clinical symptoms. Immunomodulatory agents such as steroids, plasmapheresis and intravenous immunoglobulin, seem to offer substantial improvement. PMID- 12538970 TI - [Tracheal intubation in a case of idiopathic pulmonary fibrosis using remifentanil and propofol without muscle relaxants]. AB - The anesthetic management of a 58 year-old-male suffering from idiopathic pulmonary fibrosis associated with a previous experience of altered dose-response relationship to muscle relaxants is reported. He underwent a gastroendoscopic surgical procedure. After intravenous atropine, remifentanil 3.0 microg kg(-1) was injected over 90 sec. Sixty seconds after the start of remifentanil bolus dose, propofol 2.0 mg kg(-1) was injected and intubation was performed 1 min after the administration of propofol. Anesthesia was maintained by N(2)O, oxygen, sevoflurane (0.5-0.8%) and remifentanil titrated infusion (mean dose of 0.15 mg kg(-1) min(-1)) under spontaneous respiration or assisted ventilation, if the patient was apnoic. The intubating conditions were good, a 30 per cent reduction in MAP was observed after propofol administration. The patient regained consciousness 5 minutes after the end of remifentanil administration and his trachea was extubated without any troubles. Recovery was uneventful without the need of long-term intensive therapy. PMID- 12538971 TI - Active euthanasia?. No, thanks. PMID- 12538973 TI - Lewis and Clark, pioneers, and the American dream. PMID- 12538974 TI - Beyond Flatland. PMID- 12538975 TI - The ABCs of LTX for BAC. PMID- 12538976 TI - Nothing is fair or good alone. PMID- 12538977 TI - The solitary pulmonary nodule: can we afford to watch and wait? PMID- 12538978 TI - Defining the role of anastomotic devices in coronary bypass surgery. PMID- 12538979 TI - Attenuation of postischemic myocardial injury by cariporide. PMID- 12538980 TI - The unwanted response to cardiac surgery: time for a reappraisal? PMID- 12538981 TI - Application of helical computed tomographic angiography with differential color imaging three-dimensional reconstruction in the diagnosis of complicated congenital heart diseases. PMID- 12538982 TI - New approach for complete endocardiectomy in left ventricular endomyocardial fibrosis. PMID- 12538983 TI - Successful use of a miniaturized bypass system with the DeltaStream extracorporeal rotary blood pump. PMID- 12538984 TI - Pulmonary transplantation for advanced bronchioloalveolar carcinoma. AB - BACKGROUND: No effective therapy is currently available for the diffuse stage of bronchioloalveolar carcinoma. OBJECTIVE: We tested the hypothesis that total lung replacement with standard lung transplantation techniques would provide curative therapy. METHODS: Nine patients aged 31 to 58 years with bronchioloalveolar carcinoma were entered in the study. Five patients initially had bilateral diffuse tumor. Four patients had recurrence in the contralateral lung after pulmonary resection. RESULTS: Between 1993 and 1998, all 9 patients underwent transplantation (2 single-lung and 7 bilateral transplants, 1 reoperative single lung transplant, and 1 reoperative bilateral transplant). Two patients had mediastinal node metastasis (level 7) at the time of transplantation, and 1 of these had a frankly invasive adenocarcinoma. Of the 8 patients with pure bronchioloalveolar carcinoma, 6 had recurrent pulmonary tumor after transplantation. In 2 of these patients the tumor was localized and could be resected with left lower lobectomy in one case and left pneumonectomy in the other. One is alive 89 months after transplantation; the other died 82 months after transplantation. Four other patients had a diffuse pattern of pulmonary recurrence. Two died of progressive pulmonary failure; 1 of these had retransplantation with recurrence. A third patient died of cerebral edema shortly after bilateral retransplantation. The other patient is alive with recurrence 39 months after transplantation and has bronchiolitis obliterans. Two patients without recurrence are well with unrestricted performance levels 87 and 76 months after transplantation. CONCLUSIONS: Transplantation produces a powerful palliative outcome in patients with advanced bronchioloalveolar carcinoma, but the recurrence rate is high. Transplantation for this indication remains controversial. PMID- 12538985 TI - Use of two recipient lists for adults requiring heart transplantation. AB - OBJECTIVE: An alternate (second) adult recipient list was used to match excluded potential recipients with nonstandard donor hearts that would otherwise be unused. METHODS: The only absolute criterion for entering the alternate recipient list was age: 65 years old before 1998 and 70 years old after that. Group I consisted of alternates who underwent transplantation, and group II consisted of 401 contemporaneous recipients. Hearts were first offered to regularly listed patients. At least one of the following donor risks accounted for allocation to an alternate: coronary artery disease, reused transplanted heart, high-risk behavior, hepatitis seropositivity, decreased left ventricular ejection fraction, high inotropic requirement, left ventricular hypertrophy, age older than 55 years plus another risk, and small donor with no other matches. RESULTS: Of 102 alternates, 82 were listed were because of age. After a median wait of 107 days, 62 alternates underwent transplantation. Median alternate recipient age was 67 years (vs 54 years, P <.001). Median donor age was 45 years (vs 31 years, P <.001). Survival for alternates at 90 days was 82% (vs 91%, P =.04). Significant recipient predictors of early mortality on multivariable analysis (n = 463) were previous cardiac surgery (odds ratio 2.74, 95% confidence interval 1.37-5.48) and renal dysfunction (odds ratio 1.39, 1.10-176). Alternate listing did not independently predict early or late mortality. Late (>90 days) death rates per 1000 person-months were 4.3 and 3.6 for groups I and II (relative risk 1.2, 0.62 2.36). CONCLUSIONS: Use of two adult recipient lists facilitated allocation of unused donor organs. Satisfactory long-term survival supports the use of an alternate recipient list. PMID- 12538986 TI - Baboons undergoing orthotopic concordant cardiac xenotransplantation surviving more than 300 days: effect of immunosuppressive regimen. AB - OBJECTIVE: We reviewed long-term survival among hosts in 3 consecutive series of a rhesus monkey-baboon orthotopic cardiac xenotransplantation model with reference to host immune response, including the effectiveness in preventing rejection and limiting toxicity concerning infection, to evaluate specific immunosuppressive regimens for long-term outcomes. METHODS: Six juvenile baboons surviving more than 300 days after transplantation were reviewed. Regimen A consisted of splenectomy, FK506, methotrexate, and antilymphocyte globulin. Regimen B consisted of pretransplantation and chronic maintenance with cyclosporine A (INN: ciclosporin), methotrexate, and antithymocyte globulin. Regimen C was the same as regimen B plus pretransplantation total lymphoid irradiation and intraoperative donor bone marrow cell infusion. Rejections were detected by means of echocardiography. RESULTS: Long-term survivors in 3 groups were followed for a range of 332 to 515 days (mean, 436 days). Rejection frequency in regimens A, B, and C was 0.35, 0.58, and 0.18 per month, and rescue therapy days were 23 (4.8%), 123 (9.5%), and 20 (2.4%), respectively (P <.0001). Infection frequency was 0.58, 0.56, and 0.19 per month, and therapy days were 192 (38.2%), 164 (12.6%), and 7 (0.9%), respectively (P <.0001). Concerning the host immune response, interleukin 2-activated T cells of all groups during rejection free periods showed lower numbers compared with those of control animals (P <.0005), and regimen C was the lowest among 3 groups (P <.01). The production of xenoantibody was sufficiently attenuated in all groups. CONCLUSION: Regimen C leads to long-term survival with fewer rejection and infection episodes by means of suppression of the interleukin 2 pathway and xenoantibody production. PMID- 12538987 TI - Suitable shunt size for regulation of pulmonary blood flow in a canine model of univentricular parallel circulations. AB - OBJECTIVE: We examined the influence of shunt size on regulation of the pulmonary blood flow in a canine model of a univentricular heart because specific guidelines regarding suitable shunt size in the Norwood operation remain undetermined. METHODS: Beagle dogs (n = 8) 3 to 7 months old and weighing 3.0 to 5.0 kg were used. Atrial septectomy and patch closure of the tricuspid valve were performed, and a systemic-pulmonary arterial shunt was created by interposing a 3.5- or 4.0-mm graft between the right subclavian artery and main pulmonary artery. After cardiopulmonary bypass, hemodynamic variables including pulmonary and systemic blood flow were measured consecutively according to physiologically respiratory manipulations. The ratio of shunt size to body weight ranged from 0.80 to 1.33 mm/kg (1.08 +/- 0.16 mm/kg). RESULTS: Each dog with a ratio of shunt size to body weight of 0.8 to 1.1 showed significant negative correlation between the pulmonary/systemic blood flow ratio and arterial Pco(2), but those with a ratio of shunt size to body weight of 1.1 to 1.4 did not. Consequently each dog with a ratio of shunt size to body weight of 0.8 to 1.0 got adequate systemic flow, whereas a ratio of 1.0 to 1.4 resulted in inadequate systemic flow and acidic status. Similar phenomena were shown with the grouped data on relationship between the pulmonary/systemic blood flow ratio and inspired oxygen fraction. CONCLUSIONS: These findings imply that when the ratio of shunt size to body weight is 0.8 to 1.1, the pulmonary/systemic blood flow ratio is controllable by physiologic respiratory manipulations. Larger shunts make pulmonary blood flow excessive and uncontrollable. We recommend that a ratio of shunt size to body weight of 0.9 to 1.0 be considered a useful index for suitable systemic-pulmonary arterial shunt in the Norwood operation. PMID- 12538988 TI - Primary thoracoscopic treatment of empyema in children. AB - OBJECTIVE: The optimal treatment of pediatric empyema remains controversial. The objective of this study is to compare the use of conventional management versus primary thoracoscopic drainage and decortication in children with empyema. METHODS: Conventional management has consisted of chest drain insertion under general anesthesia plus intravenous antibiotics. Thoracoscopic drainage and decortication has consisted of primary thoracoscopic drainage and decortication plus antibiotics. The clinical course of 54 patients treated conventionally between 1989 and 1997 was compared with that of 21 patients treated by means of thoracoscopic drainage and decortication between September 2000 and September 2001. RESULTS: Results of the study demonstrated that patients in the drainage decortication group had fewer invasive interventions per patient than those in the conventional management group (1.0 vs 1.26). Patients undergoing thoracoscopic drainage and decortication also had significantly shorter durations of intravenous antibiotic therapy (7.6 +/- 1.2 vs 18.2 +/- 7.5 days), chest tube drainage (4.0 +/- 0.5 vs 10.2 +/- 6.1 days), and hospital stays (7.4 +/- 0.8 vs 15.4 +/- 7.4). Moreover, there were no open thoracotomies and decortications in the thoracoscopic drainage and decortication group, whereas in the conventional management group 39% (21/54) of patients underwent an open procedure. CONCLUSION: Although the 2 groups were not prospectively randomized and they were treated in different time periods, the results of this study support the use of thoracoscopic surgery as the primary therapeutic modality in children presenting with pleural empyema. This strategy appears to offer significant benefits over conventional treatment in terms of duration of treatment and the need for more invasive surgery. PMID- 12538989 TI - Hyperbaric oxygen as a chemotherapy adjuvant in the treatment of metastatic lung tumors in a rat model. AB - OBJECTIVES: The objectives of the study were to test the hypothesis that hyperbaric levels of oxygen enhance the sensitivity of a sarcoma cell line to doxorubicin (Adriamycin) both in vitro and in vivo in a rat model of pulmonary metastases and to test the feasibility of arterialization of mixed venous blood by direct injection of aqueous oxygen into the pulmonary artery in a rat model. METHODS: Rat sarcoma (MCA-2) cells were incubated in the presence of increasing concentrations of doxorubicin (0.1-2.0 micromol/L). A dose-dependent toxicity relationship at 12 hours of treatment was examined with and without pretreatment with hyperbaric oxygen (3.7 atm absolute for 1.5-3.5 hours). In vivo, Sprague Dawley rats (n = 24) were injected intravenously with 10(6) MCA-2 cells, and the lung tumors were allowed to mature for 14 days. At that time the animals were divided into four groups: control (no treatment), doxorubicin at 2 mg/kg, hyperbaric oxygen (oxygen at 2 atm absolute for 30 minutes), and hyperbaric oxygen plus doxorubicin. Seven days after treatment, the numbers of lung nodules were counted and the lung weights were determined. In additional rats (n = 7), aqueous oxygen (1 mL oxygen/g saline solution) was infused into the pulmonary artery to determine whether arterialization of mixed venous blood was comparable to pulmonary artery oxygenation with a hyperbaric chamber (n = 7). RESULTS: Hyperbaric oxygen plus doxorubicin produced significantly greater cytolysis of MCA-2 cells (P <.01) than did doxorubicin alone. Hyperbaric oxygen plus doxorubicin also significantly decreased the number of lung metastases and the lung weight relative to doxorubicin alone (P <.01 and P <.01, respectively). The feasibility of arterialization of mixed venous blood (>100 mm Hg) with aqueous oxygen infusion was demonstrated. CONCLUSIONS: Hyperbaric oxygen enhanced the chemotherapeutic effect of doxorubicin both in cell culture and in the rat model. Aqueous oxygen infusion can be used to oxygenate mixed venous blood at levels similar to those obtained with the use of a hyperbaric chamber. PMID- 12538990 TI - Efficacy and safety of tracheobronchoplasty after induction therapy for locally advanced lung cancer. AB - OBJECTIVES: Patients receiving induction therapy may have increased risk of morbidity and mortality after surgery. We retrospectively evaluated the influence of preoperative treatment in patients who underwent sleeve resection for lung cancer. METHODS: A series of 48 consecutive patients who underwent sleeve resection with a telescoping anastomosis was retrospectively analyzed. A sleeve lobectomy and pneumonectomy were performed in 41 and 7 patients, respectively. Twenty patients received preoperative induction therapy; of them, 16 received induction chemoradiotherapy and 4 received only chemotherapy. Twenty-eight patients underwent the procedure without adjuvant therapy. RESULTS: The telescopic procedure was performed by placing sutures around the proximal and distal portions of the bronchial cartilage without wrapping the anastomosis. Among the 20 patients who received induction therapy, pulmonary angioplasty was performed in 5 and chest wall resection was performed in 3. Seven of these 20 patients (35%) had postoperative complications. Among the 28 patients without preoperative adjuvant therapy, pulmonary angioplasty was performed in 3, diaphragmatic resection was performed in 1, and chest wall resection was performed in 1. Three of these 28 patients (11%) had postoperative complications. Complications relating to the anastomosis occurred in 1 patient (5.0%) who received induction therapy; however, no operative deaths occurred. Bronchoscopic examinations demonstrated that mucosal healing was prolonged in patients who underwent induction therapy. CONCLUSION: Induction therapy did not significantly affect morbidity or mortality among patients who underwent sleeve resection. PMID- 12538991 TI - Intraoperative lavage cytologic analysis of surgical margins in patients undergoing limited surgery for lung cancer. AB - BACKGROUND: One of the unfavorable recurrent patterns after limited surgery for lung cancer is local failure, especially at the surgical margin in the pulmonary parenchyma. To prevent this failure, we preliminarily introduced a novel intraoperative lavage cytologic technique to check surgical margin status for limited surgery. In this study we analyzed the clinical utility of this technique with a larger number of patients under long-term follow-up. METHODS: A total 112 consecutive lung cancer lesions prospectively treated by limited surgery with the intraoperative lavage cytologic technique between October 1997 and August 2000 were reviewed through a median follow-up period of 27 months. RESULTS: Eleven lesions (10%) showed cytologically positive results in the attempted surgery on the surgical margin. The positive result rate was significantly higher for lesions with more advanced stage, compromised indication, incurability, and larger size. Surgical modes were converted intraoperatively for 4 lesions; in the other 7 lesions no conversion was performed because of certain disadvantages. Local recurrence in the surgical margin occurred in a total of 4 lesions, including 3 for which the operative mode was unconverted and 1 lesion with cytologically unknown status of the surgical margin that had the mode converted, whereas there were no local recurrences in the surgical margins among the lesions with final cytologically negative results. CONCLUSION: Cytologically negative results of examination of the surgical margin by the technique of intraoperative lavage cytologic in limited surgery for lung cancer may be predict lack of local recurrence in the surgical margin. This intraoperative cytologic technique is clinically useful in checking for complete resection of this primary disease. PMID- 12538993 TI - Appropriate timing of surgical intervention after transmural acute myocardial infarction. AB - OBJECTIVE: Recommended timing of coronary revascularization after transmural acute myocardial infarction ranges from immediate surgical intervention to repair 4 weeks after infarction. Such wide variation has created a dilemma in the management of these patients. The objective of this study was to delineate the optimal timing of revascularization after transmural acute myocardial infarction in a large and contemporary patient population. METHODS: We performed a retrospective multicenter analysis of 32,099 patients who underwent coronary artery bypass grafting as the sole procedure after transmural myocardial infarction between 1991 and 1996 by 179 surgeons at 33 hospitals in New York State. RESULTS: Overall hospital mortality for all patients who underwent coronary revascularization with a history of transmural myocardial infarction was 3.3%. Hospital mortality decreased with increasing time interval between revascularization and transmural acute myocardial infarction: 14.2%, 13.8%, 7.9%, 3.8%, 2.9%, and 2.7% for less than 6 hours, 6 hours to 1 day, 1 to 3 days, 4 to 7 days, 7 to 14 days, and greater than 15 days, respectively. Multivariate analyses of 43 potential risk factors suggests that revascularization within 3 days of transmural acute myocardial infarction is independently associated with mortality. CONCLUSIONS: Coronary revascularization within 3 days of a transmural acute myocardial infarction might be an added risk for mortality. In the absence of absolute indications for emergency surgical intervention, such as structural complications and ongoing ischemia, a 3-day waiting period before surgical revascularization should be considered. PMID- 12538992 TI - Effect of preoperative delay on prognosis for patients with early stage non-small cell lung cancer. AB - OBJECTIVE: Screening for lung cancer will discover many nodules of indeterminate pathology. Observation has the theoretic risk of permitting dissemination of a localized cancer and worsening prognosis, whereas immediate evaluation of benign conditions generates morbidity and cost. This study was conducted to assess the effect of delay in surgical intervention on survival for patients with early stage non-small cell lung cancer. METHODS: Records for patients with resected pathologic stage I and II non-small cell lung cancer (1989-1999) were abstracted for patient age, race, sex, medical history, date of presentation, date and type of surgical treatment, pathologic stage, and date of death or last follow-up. Kaplan-Meier survival analysis was performed to test for the effect of delay (time from presentation to surgical intervention) on survival. RESULTS: Eighty four patients were identified. Median age was 66 years, median preoperative interval was 82 days (range, 1-641 days), and median follow-up was 3.3 years (range, 5 days-11.9 years). Median survival was 3.7 years. Overall 5-year survival was 40%; disease-specific 5-year survival was 63%. Log-rank analysis of the effect of delay on overall survival generated a P value of.54, with an estimated hazard ratio for a 90-day delay of 1.06 (95% confidence interval, 0.87 1.30). CONCLUSIONS: For this population, we were unable to detect a significant effect of delay on prognosis. Although these results suggest that the risk of judicious observation of indeterminate pulmonary nodules might be low, the 95% confidence interval is broad. Larger sample sizes are needed to reach definitive conclusions. PMID- 12538994 TI - Continuous retrograde blood cardioplegia is associated with lower hospital mortality after heart valve surgery. AB - OBJECTIVE: Myocardial preservation studies comparing blood and crystalloid cardioplegia techniques were almost exclusively performed on patients undergoing coronary bypass, and they were unable to show a difference in hospital mortality. We investigated possible factors, including cardioplegia techniques, influencing hospital mortality in patients undergoing cardiac valve surgery. METHODS: We evaluated hospital mortality in 1098 consecutive patients undergoing cardiac valve surgery by using a multivariate logistic regression with propensity score balancing of the groups. In 25% of the patients, multiple valve or Bentall procedures were performed, and in 46% of all patients, coronary bypass grafting was associated with valve surgery. A first cohort of 504 consecutive patients were operated on by using single-shot antegrade cold crystalloid cardioplegia, and a second cohort of 594 patients were operated on by using continuous retrograde cold blood cardioplegia. RESULTS: After correction for patient-related and operative risk factors, lower hospital mortality was found in patients who received retrograde blood cardioplegia (P =.020). The odds ratio of in-hospital death when using blood cardioplegia was 0.44 (95% confidence interval, 0.22 0.88). Further predictors of hospital mortality were age, advanced New York Heart Association functional class, cardiopulmonary bypass time, reoperation, active endocarditis, and renal failure. CONCLUSIONS: This study shows that continuous retrograde blood cardioplegia is associated with lower hospital mortality in heart valve operations. PMID- 12538995 TI - Total replacement of the ascending aorta without circulatory arrest. AB - OBJECTIVE: If the aortic arch is clamped between the brachiocephalic trunk and the left common carotid artery and the brachiocephalic trunk is also clamped, the total ascending aorta can be replaced without circulatory arrest. Candidates for this technique need to have the following characteristics: preoperative demonstration of Willis polygon patency by means of transcranial Doppler ultrasonography, preoperative computed tomographic scan that shows no calcification in the arch or in the brachiocephalic trunk, and preoperative or intraoperative evidence of separate origins of the brachiocephalic trunk and the left common carotid artery. METHODS: In three different institutions (with different time frames in each) from December 2000 to December 2001, the possibility of replacing the total ascending aorta without circulatory arrest was evaluated in 14 cases. In 10 of them it was possible (feasibility of 71.4%). During the procedure continuous electroencephalographic monitoring was performed. RESULTS: No patient died in the early or midterm follow-up. None of the patients had any cerebral complications. The electroencephalographic activity was normal during the period of brachiocephalic trunk occlusion. In 5 patients a postoperative spiral computed tomographic scan showed good surgical results. CONCLUSIONS: The technique described here allows replacement of the total ascending aorta without circulatory arrest in selected cases. Furthermore, the evidence of lack of right cerebral hypoperfusion in all the procedures we performed allows future exploration of the possibility of avoiding cerebral monitoring. PMID- 12538996 TI - Video-assisted multivessel revascularization through a left anterior small thoracotomy approach with the Symmetry Aortic Connector System. AB - OBJECTIVE: We sought to demonstrate the applicability of video-assisted multivessel revascularization through a left anterior small thoracotomy approach with the use of the Symmetry Aortic Connector System (St Jude Medical Anastomotic Technology Group, Inc, St Paul, Minn) as an alternative to the standard median sternotomy approach and to evaluate predischarge angiographically documented graft patency. METHODS: From October 2001 through February 2002, a total of 15 patients with triple-vessel disease were operated on through a left anterior small thoracotomy approach with video-assisted port-access construction of proximal aorta-to-saphenous vein anastomoses with the Symmetry Aortic Connector System and cardiopulmonary bypass with femoral cannulation and without cardioplegic arrest. There were 9 male and 6 female subjects with a mean age of 68.3 +/- 3.6 years and an average ejection fraction of 55.8% +/- 19.6%. Subject inclusion criteria consisted of female sex (initially but not throughout the study), coronary artery reoperations, and sternal bone disease. Subject exclusion criteria consisted of an age younger than 65 years, extensive atheromatous plaques in the ascending aorta, and aortoiliac occlusive disease. All but 1 patient underwent angiographic patency evaluation before discharge. RESULTS: Fifteen operations were performed successfully without any deaths. Twenty-nine sutureless proximal anastomoses were performed, with an average of 3.13 +/- 0.62 distal anastomoses per patient. Eleven (73%) patients underwent a fast track protocol with extubation in the operating room. We did not observe any instances of low cardiac output syndrome, stroke, renal insufficiency, wound complication, or perioperative myocardial infarction. A single episode of atrial fibrillation occurred in this group. Angiographic assessment of 44 bypass grafts and target arteries was performed, and 86% of those examined were widely patent (FitzGibbon score A). CONCLUSIONS: We have demonstrated a potential advantage of the sutureless Symmetry Aortic Connector System as a suitable approach that affords minimal access. Video-assisted multivessel revascularization through a left anterior small thoracotomy approach with an automated mechanical anastomosis device is particularly useful in patients undergoing coronary artery bypass reoperations or those at risk of poor sternal healing or infection. This approach seems to be a safe alternative to standard median sternotomy. PMID- 12538997 TI - Mechanism of higher incidence of ischemic mitral regurgitation in patients with inferior myocardial infarction: quantitative analysis of left ventricular and mitral valve geometry in 103 patients with prior myocardial infarction. AB - OBJECTIVE: The mechanism of higher incidence of ischemic mitral regurgitation in patients with inferior compared with anterior myocardial infarction despite less global left ventricular remodeling and dysfunction is controversial. We hypothesized that inferior myocardial infarction causes left ventricular remodeling, which displaces posterior papillary muscle away from its normal position, leading to ischemic mitral regurgitation. METHODS: In 103 patients with prior myocardial infarction (61 anterior and 42 inferior) and 20 normal control subjects, we evaluated the grade of ischemic mitral regurgitation on the basis of the percentage of Doppler jet area, left ventricular end-diastolic and end systolic volumes, midsystolic mitral annular area, and midsystolic leaflet tethering distance between papillary muscle tips and the contralateral anterior mitral annulus, which were determined by means of quantitative echocardiography. RESULTS: Global left ventricular dilatation and dysfunction were significantly less pronounced in patients with inferior myocardial infarction (left ventricular end-systolic volume: 52 +/- 18 vs 60 +/- 24 mL, inferior vs anterior infarction, P<.05; left ventricular ejection fraction: 51% +/- 9% vs 42% +/- 7%, P <.0001). However, the percentage of mitral regurgitation jet area and the incidence of significant regurgitation (percentage of jet area of 10% or greater) was greater in inferior infarction (percentage of jet area: 10.1% +/- 7.5% vs 4.4% +/- 7.0%, P =.0002; incidence: 16/42 (38%) vs 6/61 (10%), P <.0001). The mitral annulus (area = 8.2 +/- 1.2 cm2 in control subjects) was similarly dilated in both inferior and anterior myocardial infarction (9.7 +/- 1.7 vs. 9.5 +/- 2.3 cm2, no significant difference), and the anterior papillary muscle-tethering distance (33.8 +/- 2.6 mm in control subjects) was also similarly and mildly increased in both groups (35.2 +/- 2.4 vs 35.2 +/- 2.8 mm, no significant difference). However, the posterior papillary muscle-tethering distance (33.3 +/- 2.3 mm in control subjects) was significantly greater in inferior compared with anterior myocardial infarction (38.3 +/- 4.1 vs 34.7 +/- 2.9 mm, P =.0001). Multiple stepwise regression analysis identified the increase in posterior papillary muscle-tethering distance divided by body surface area as an independent contributing factor to the percentage of mitral regurgitation jet area (r2 = 0.70, P <.0001). CONCLUSIONS: It is suggested that the higher incidence and greater severity of ischemic mitral regurgitation in patients with inferior compared with anterior myocardial infarction can be related to more severe geometric changes in the mitral valve apparatus with greater displacement of posterior papillary muscle caused by localized inferior basal left ventricular remodeling, which results in therapeutic implications for potential benefit of procedures, such as infarct plication and leaflet or chordal elongation, to reduce leaflet tethering. PMID- 12538998 TI - Effect of diabetes on early and late survival after isolated first coronary bypass surgery in multivessel disease. AB - OBJECTIVE: Diabetes has not yet been investigated as a risk factor for early and late cardiac-related death. METHODS: Patients operated on from January 1988 to December 1999 were considered; 767 were diabetic (group D) and 2593 were nondiabetic (group ND). Patients with preoperative hemodynamic deterioration were excluded. Early (30-day) mortality (any causes and cardiac causes) was evaluated with univariate analysis and stepwise logistic regression. Ten-year actuarial freedom from death of any cause and cardiac death was also assessed with univariate and Cox analyses. RESULTS: Early mortality was 2.2% (group D, 3.3%; group ND, 1.9%; P =.023). Early cardiac mortality was 1.3% (group D, 2.2%; group ND, 1.1%; P =.0016). Diabetes was an independent risk factor only for cardiac death and not for death of any cause. Five-year survival was 93.5% +/- 0.5% (group D, 92.5% +/- 1.1%; group ND, 93.9% +/- 0.6%; P =.0304). Diabetes was not an independent risk factor. Five-year freedom for cardiac death was 96.3% +/- 0.4% (group D, 94.9% +/- 0.9%; group ND, 96.6% +/- 0.4%; P =.0155). Diabetes was an independent risk factor. However, if only the patients who survived the first 30 days are considered, diabetes disappears as a risk factor (5-year freedom for cardiac death, 97.8% +/- 0.3%; group D, 97.3% +/- 0.8%; group ND, 97.9% +/- 0.4%; P = 0.2389). CONCLUSIONS: Diabetes is an independent risk factor for early cardiac death only. Long-term survival in patients who survive the first 30 days is not statistically significantly different for diabetic and nondiabetic patients. In fact, the rates appear very similar. PMID- 12538999 TI - Blood cardioplegia supplementation with the sodium-hydrogen ion exchange inhibitor cariporide to attenuate infarct size and coronary artery endothelial dysfunction after severe regional ischemia in a canine model. AB - BACKGROUND: Activation of the sodium-hydrogen ion exchange mechanism results in accumulation of intracellular calcium through the sodium-calcium ion antiport mechanism. Administration of a sodium-hydrogen ion exchange inhibitor before or during ischemia attenuates myocardial ischemia and reperfusion injury. However, the cardioprotection exerted by sodium-hydrogen ion exchange inhibitors as adjuncts to cardioplegia without perioperative administration has not been tested in a model of surgical reperfusion of acute coronary occlusion with cardiopulmonary bypass. This study tested the hypothesis that sodium-hydrogen ion exchange inhibitor-supplemented blood cardioplegia would reduce postcardioplegia injury after severe regional ischemia. METHODS: In anesthetized open-chest dogs, the left anterior descending coronary artery was occluded for 75 minutes, after which total cardiopulmonary bypass was initiated. After crossclamping, cold (4 degrees C) antegrade blood cardioplegia was delivered every 20 minutes for a total of 60 minutes of cardioplegic arrest. In 8 dogs, the blood cardioplegic solution was unsupplemented (vehicle group), whereas in 8 others the solution was supplemented with the sodium-hydrogen ion exchange inhibitor cariporide (10 micro mol/L, cariporide group). RESULTS: In the in vitro studies, the direct effects of cariporide on neutrophil function were determined. Isolated canine neutrophils were stimulated by platelet activating factor. Cariporide attenuated superoxide anion production in a concentration-dependent manner, with no appreciable effect at 10 micro mol/L (the concentration used in blood cardioplegia) and a peak effect at 100 micro mol/L. In the in vivo cardiopulmonary bypass model, infarct size was significantly (P <.05) smaller in the cariporide group than in the vehicle group (22.4% +/- 3.5% vs 40.1% +/- 5.1% of area at risk), although there were no group differences in postischemic regional wall motion after 2 hours of reperfusion (0.1% +/- 0.9% vs -0.2% +/- 0.3% systolic shortening). Transmural myocardial edema in the area at risk was significantly decreased in the cariporide group (80.6% +/- 0.5%) relative to the vehicle group (83.1% +/- 0.6%). Myeloperoxidase activity in the area at risk, an index of neutrophil accumulation, was significantly lower in the cariporide group than in the vehicle group (4.7 +/- 0.9 absorbence units/[min. g tissue] vs 10.3 +/- 2.3 absorbence units/[min. g tissue]). In isolated postischemic left anterior descending coronary artery rings, maximum relaxation in response to the endothelium dependent vasodilator acetylcholine was significantly greater in the cariporide group than in the vehicle group (77.5% +/- 7.4% vs 51.4% +/- 8.0%), whereas smooth muscle relaxation in response to nitroprusside was comparable between groups. CONCLUSION: In this canine model, supplementation of blood cardioplegia with cariporide, a sodium-hydrogen ion exchange inhibitor, reduced infarct size, attenuated neutrophil accumulation in the area at risk, and reduced postischemic coronary artery endothelial dysfunction without directly inhibiting neutrophil activity. Cariporide as an adjunct to blood cardioplegia without perioperative administration attenuated surgical ischemia-reperfusion injury in jeopardized myocardium. PMID- 12539000 TI - Nuclear factor kappaB mediates a procoagulant response in monocytes during extracorporeal circulation. AB - OBJECTIVE: The objective of this study was to examine the mechanism of procoagulant activity and inhibition in whole blood during extracorporeal circulation. METHODS: In this study we examine the development of procoagulant activity and monocyte activation in heparinized whole blood passing through a closed circuit consisting of a pump and silicone envelope membrane oxygenator for 6 hours. RESULTS: Anaphylatoxins, C3a and C5a, determined by means of enzyme linked immunosorbant assay, appeared in the blood within 30 minutes of circulation. Circulated blood developed a marked potential for coagulation demonstrated in a 1-step clotting assay that reached maximal activity by 4 hours of circulation. This procoagulant activity was neutralized by anti-tissue factor antibody, suggesting a prominent role for the extrinsic pathway in pump-induced intravascular coagulation. Isolation of monocytes from circulated blood revealed that tissue factor expression is upregulated on the cell surface. Furthermore, we observed nuclear factor kappaB nuclear translocation in monocytes from blood passing through the circuit, suggesting that tissue factor expression was due to monocyte stimulation and transcriptional activation of the tissue factor gene. Tissue factor expression resulted in an approximately 30-fold increase in thrombin generation. Monocyte nuclear factor kappaB activation, monocyte tissue factor expression, thrombin generation, and the procoagulant activity of blood in extracorporeal circulation were all blocked by the proteasome inhibitor MG132. CONCLUSIONS: We conclude that intravascular tissue factor expression during extracorporeal circulation of blood is due to nuclear factor kappaB-mediated activation of monocytes (possibly by complement), which can be controlled pharmacologically. PMID- 12539001 TI - Phosphodiesterase type 4 inhibition of activated polymorphonuclear leukocytes in a simulated extracorporeal circulation model. AB - OBJECTIVES: Cardiopulmonary bypass is associated with a systemic inflammatory response syndrome and the risk of multiorgan injuries mediated by activated polymorphonuclear leukocytes. Phosphodiesterase type 4 is the predominant phosphodiesterase isozyme in polymorphonuclear leukocytes and plays a key role in the regulation of polymorphonuclear leukocyte activation. The aim of this study was to examine the effect of rolipram, a selective phosphodiesterase type 4 inhibitor, on the functional changes of polymorphonuclear leukocytes by using simulated extracorporeal circulation. METHODS: Simulated extracorporeal circulation was established by recirculating heparinized human blood for 120 minutes on a membrane oxygenator with and without 10 micro mol/L rolipram. F actin content and L-selectin and CD11b expression of polymorphonuclear leukocytes were measured by means of flow cytometry. Polymorphonuclear leukocyte deformability was evaluated with a microchannel array flow analyzer that had a similar diameter as the capillaries. Polymorphonuclear leukocyte elastase was measured with an enzyme immunoassay. RESULTS: Rolipram reduced the increase of F actin content of polymorphonuclear leukocytes and the increase of transit time of 100 micro L of blood sample through a microchannel. Rolipram reduced the increase of CD11b expression and the decrease of L-selectin expression of polymorphonuclear leukocytes. Rolipram reduced the release of elastase from polymorphonuclear leukocytes. CONCLUSION: Rolipram inhibited the deformability change mediated by F-actin assembly, the changes in adhesion molecules, and the release of elastase from activated polymorphonuclear leukocytes in simulated extracorporeal circulation. This study suggests that phosphodiesterase type 4 inhibition could be a feasible therapeutic strategy to prevent the exaggerated inflammatory response related to cardiopulmonary bypass. PMID- 12539002 TI - Nitrotyrosine and 8-isoprostane formation indicate free radical-mediated injury in hearts of patients subjected to cardioplegia. AB - OBJECTIVE: Myocardial ischemia and reperfusion induced by cardioplegic arrest subjects the heart to free radical-mediated stress. The purpose of our study was to investigate the effect of cardioplegia-induced ischemia and reperfusion on myocardial formation and distribution of (1) nitrotyrosine as an indicator for peroxynitrite-mediated tissue injury resulting from increased nitric oxide release and (2) 8-isoprostane as an indicator for oxygen-derived free radical mediated lipid peroxidation. METHODS: In 10 patients undergoing coronary artery operations (64 +/- 6 [mean +/- SD] years, 3 women and 7 men) subjected to cardiopulmonary bypass and intermittent cold blood cardioplegia, we collected transmural left ventricular biopsy specimens before and at the end of cardiopulmonary bypass. Specimens were cut at 10 micro m and subjected to immunocytochemical staining against the nitric oxide-producing enzyme constitutive nitric oxide synthase, cyclic guanosine monophosphate (intracellular second messenger of nitric oxide), nitrotyrosine, and 8-isoprostane by using polyclonal antibodies. For global left ventricular function determination, we measured the fractional area of contraction using transesophageal echocardiography. RESULTS: Nitric oxide synthase activity in cardiac myocytes increased from 34 +/- 10 gray units before cardiopulmonary bypass to 47 +/- 12 gray units at the end of bypass (P =.015), and all hearts showed increased cyclic guanosine monophosphate content in both myocytes and endothelial cells at the end of bypass. The number of nitrotyrosine-positive capillaries increased from 36 +/- 29/mm(2) before bypass to 82 +/- 47/mm(2) at the end of bypass (P =.012), and 8 isoprostane-positive capillaries increased from 92 +/- 72/mm(2) before bypass to 209 +/- 108/mm(2) at the end of bypass (P =.005). The fractional area of contraction was 53% +/- 12% before bypass and 56% +/- 12% after bypass (P =.47) but was slightly decreased to 45% +/- 14% at 4 hours after bypass (P =.121). CONCLUSIONS: Our data show that cardioplegia-induced myocardial ischemia and reperfusion is associated with nitrotyrosine and 8-isoprostane formation mainly in the coronary endothelium, indicating injury mediated by both peroxynitrite and oxygen-derived free radicals. Because nitric oxide synthase activation was accompanied with increased cyclic guanosine monophosphate, these data suggest that direct effects of nitric oxide on cardiac myocytes, as well as nitric oxide mediated coronary endothelial injury, might contribute to injury associated with cardioplegia and cardiopulmonary bypass. PMID- 12539003 TI - Does aprotinin influence the inflammatory response to cardiopulmonary bypass in patients? AB - OBJECTIVES: Aprotinin has been shown to have anti-inflammatory properties, but its effects on the inflammatory reaction to cardiopulmonary bypass remain controversial. This prospective, randomized, double-blind study evaluated the influence of aprotinin on various blood markers of inflammation during and after cardiopulmonary bypass. METHODS: Sixty male patients underwent coronary artery bypass grafting. The patients were randomized into 3 groups: a placebo group, a second group receiving 2,000,000 KIU of aprotinin followed by an infusion of 500,000 KIU/h and 2,000,000 KIU in the pump prime, and a third group receiving half this dosage. Measurements of tumor necrosis factor, interleukin 6, interleukin 8, interleukin 10, endotoxin, histamine, complement factors, prekallikrein, and prostaglandin D(2) were obtained at baseline, 30 minutes after study drug loading, 10 minutes after the beginning of cardiopulmonary bypass, before the end of bypass, 4 hours after bypass, and on the first and second postoperative days. RESULTS: Aprotinin had no significant effect on any of these parameters. As expected, aprotinin reduced early blood loss in both treated groups. CONCLUSIONS: These results indicate that aprotinin at doses currently used to reduce blood loss has no significant influence on the systemic inflammatory response during moderate hypothermic cardiopulmonary bypass in human subjects, as assessed by the mediators measured in this study. PMID- 12539004 TI - Surgical treatment of a primary tracheal angiosarcoma. PMID- 12539006 TI - Brucella melitensis endocarditis of ventricular septal defect patch: successful surgical treatment without replacement. PMID- 12539005 TI - Primary choriocarcinoma of the lung. PMID- 12539007 TI - The quadricuspid pulmonary valve: its importance in the Ross procedure. PMID- 12539008 TI - Aortic root replacement with Freestyle stentless valve for complex aortic root infection. PMID- 12539009 TI - Left ventricular thrombectomy in a cocaine user with normal coronary arteries. PMID- 12539010 TI - Simultaneous enlargement of the pulmonary annulus and the pulmonary cusp with a transannular patch. PMID- 12539011 TI - A ball thrombus in the left ventricle in association with Sweet syndrome. PMID- 12539012 TI - Apolipoprotein E4 increases aortic atheroma burden in cardiac surgical patients. PMID- 12539013 TI - Systemic allergic reaction to the percutaneous patent foramen ovale occluder. PMID- 12539014 TI - Right pneumonectomy and thoracoplasty followed by coronary artery bypass grafting and mitral valve replacement. PMID- 12539015 TI - Q fever endocarditis: a surgical view and a word of caution. PMID- 12539016 TI - Determinants of maximal right ventricular function: role of septal shift. PMID- 12539017 TI - Thymolipomas with myasthenia gravis in Japan. PMID- 12539018 TI - Negative pressure pulmonary edema following open reduction and internal fixation of a fractured mandible. PMID- 12539019 TI - Painless mass of the cheek. PMID- 12539020 TI - Painful, rapidly growing mass of the mandible. PMID- 12539021 TI - Osteomyelitis of the maxilla secondary to osteopetrosis: report of a case. AB - Osteomyelitis of the maxilla is extremely rare. When it occurs, there is invariably an underlying predisposing condition. We describe a 28-year-old woman whose presentation with osteomyelitis of the maxilla led to a diagnosis of generalized osteopetrosis. PMID- 12539022 TI - Orbital floor reconstruction with flexible Ethisorb patches: a retrospective long term follow-up study. AB - OBJECTIVE: The purpose of the study was to investigate whether a flexible, biodegradable material (Ethisorb) shows better long-term results with regard to diplopia, bulbus motility, and exophthalmos/enophthalmos compared to the use of lyophilized dura-patches and polydioxanone (PDS) foils. METHODS: During a period of 6 years 435 patients with an orbital fracture were investigated retrospectively. Inclusion criteria were patients with fractures of the orbital floor with a maximum size of 2 x 2 cm. Bulbus motility, exophthalmos, enophthalmos, and diplopia were investigated during a period of 2 years. RESULTS: One hundred twenty orbital floors were reconstructed by lyophilized dura-patches, 81 by PDS, and 136 by Ethisorb. An exploration without an implantation was performed in 91 patients. The long-term investigation 12 to 15 months after surgery showed an exophthalmos and enophthalmos incidence of 1%, whereas a reduced bulbus motility and diplopia were found in 5% and 4%, respectively. Fifteen to 24 months after surgery 2% of the patients had an exophthalmos and 1% had an enophthalmos. A reduction of bulbus motility was found in 4% of the patients, and diplopia was found in 3%. The use of Ethisorb resulted in a significantly lower incidence of exophthalmos 3 months after surgery compared to PDS. CONCLUSION: The low rate of acquired bulbus motility demonstrates acceptable results in using Ethisorb in the floor of the orbit. PMID- 12539023 TI - Management of epistaxis in the oral and maxillofacial surgery setting: An update on current practice. AB - Epistaxis is a common and, in most cases, benign event. Although most nosebleeds resolve spontaneously, some may be profuse and life-threatening. Severe or recurrent epistaxis can be a challenging management problem. In otolaryngologic practice, it is the most commonly seen emergency. Oral and maxillofacial surgeons will also encounter this clinical problem in varied settings. It is our aim to present an update on the contemporary management of epistaxis in maxillofacial practice. The etiology and relevant surgical anatomy are discussed. This is followed by an update on current treatment regimens in different scenarios. A stepwise algorithm for the management of epistaxis is presented. PMID- 12539024 TI - Necrotizing soft tissue infections of the head and neck: case reports and literature review. AB - A small but nevertheless important part of a surgeon's experience comprises necrotizing soft tissue infections of the head and neck. The purpose of this report is to heighten awareness of necrotizing soft tissue infections in any patient with an infection of the head and neck. The article also outlines an appropriate management strategy for use in the treatment of patients with this potentially fatal condition. Prompt diagnosis and early radical surgical debridement are significant factors in avoiding a fatal outcome in these patients. This article reviews the literature on necrotizing soft tissue infections of the head and neck and presents cases from our recent experience. PMID- 12539025 TI - A pilot study to test the efficacy of oral administration of interferon-alpha lozenges to patients with Sjogren's syndrome. AB - BACKGROUND: Complications from Sjogren's syndrome-induced xerostomia may cause a significant disruption in daily routines such as speech and eating habits. In addition, the reduction in salivary output may lead to increased incidence of caries, oral ulcers, sialadenitis, periodontal problems, and Candida infections. Interferon-alpha has been suggested as an effective drug therapy to increase salivary output. OBJECTIVE: The objective was to test the efficacy of interferon alpha lozenges in relieving the symptoms of dry mouth and dry eyes when administered orally to patients with Sjogren's syndrome in double-blind placebo controlled and open-label trials. STUDY DESIGN: Twelve patients with a diagnosis of primary Sjogren's syndrome were provided an Institutional Review Board approved, written informed-consent form. During the double-blind placebo controlled study they were randomized to receive 150 IU of interferon-alpha (8 patients) or placebo (4 patients) for 24 weeks with 6-week reevaluations. Five patients who received interferon-alpha in the double-blind placebo-controlled trial were treated in an open-label study for another 24 weeks. Whole saliva was measured during each visit, and symptoms were assessed by questionnaires and visual analog scales. The Wilcoxon signed rank test was used to detect significant changes for each variable. RESULTS: The results indicated that, by the end of 24 weeks, patients who received interferon-alpha had a statistically significant improvement in unstimulated salivary flow rate (P < .05), and statistically significant changes were observed in the median ocular dryness visual analog scale (P < .05) and oral dryness visual analog scale (P < .05). No significant changes were observed among the placebo patients. By the end of 48 weeks, medicated patients had better results than they had at the end of the double-blind placebo-controlled study and showed improvements in whole stimulated salivary flow rate, conditions of throat, and swallowing dry food. CONCLUSION: Oral administration of 150 IU interferon-alpha 3 times a day for the patients with primary Sjogren's syndrome improved saliva production, relieved symptoms of xerostomia and xerophthalmia, and was well tolerated by the patients. PMID- 12539026 TI - Increased allelic loss in toluidine blue-positive oral premalignant lesions. AB - OBJECTIVE: Recent studies have shown that a loss of chromosome regions (loss of heterozygosity [LOH]) containing known or presumptive tumor suppressor genes is predictive of the cancer risk of oral premalignant lesions. This preliminary study investigated whether the dye toluidine blue (TB) preferentially stains oral premalignant lesions with LOH. This stain has been used by clinicians to delineate dysplasia/carcinoma in the oral cavity. STUDY DESIGN: The study included 32 patients with oral lesions who underwent biopsy after the assessment of TB dye retention. A total of 39 biopsy specimens were examined (14 hyperplastic, 25 dysplastic). Fourteen of the biopsy specimens were TB-negative. The specimens were analyzed for LOH at 10 microsatellite loci on 3 chromosome arms (3p, 9p, and 17p), and the LOH results of TB-positive samples were compared with those that were negative for the tissue staining. RESULTS: TB-positive samples had a higher frequency of loss than TB-negative cases for loci on 3p (P = .013) and 17p (P = .049). In addition, more TB-positive cases showed a loss of multiple arms (>2 arms, P = .015), a pattern that has been associated with markedly increased cancer risk. CONCLUSION: The study results suggest that TB staining may help identify oral premalignant lesions with increased LOH and increased cancer risk. PMID- 12539027 TI - Reassessing the presence of Candida albicans in denture-related stomatitis. AB - OBJECTIVE: The aim of this study was to reevaluate the link between Candida albicans and denture-related stomatitis according to a modified Newton classification, which reflects the classic types of inflammation as well as the extent to which the tissue is affected. STUDY DESIGN: Two groups of denture wearers were evaluated for denture-related stomatitis. The presence and number of yeasts on the dentures, the identification of the Candida species present, and the amount of plaque coverage were determined. Putative risk factors were included in the study. Relations between these variables and stomatitis were analyzed statistically. RESULTS: According to Newton's classification, the presence of yeast on the denture was not linked to whether subjects had stomatitis. But with our classification, higher prevalence of yeast carriers, yeast colony number, and plaque coverage were found on the dentures of individuals with the most extensive inflammation, regardless of Newton type. Among risk factors evaluated, wearing dentures at night and smoking were associated with the most extensive inflammation. We also demonstrated that the presence of C albicans as well as the cohabitation of different Candida species was more frequent in denture-related stomatitis. The differences were statistically significant. CONCLUSIONS: Statistical analysis of microbiologic data from different denture-related stomatitis categories according to our modified classification showed that the presence of yeast on dentures was significantly associated with the extent of the inflammation, rather than with the Newton type. Our findings suggest that the inflammatory process of stomatitis favors the colonization of Candida. These results could have new implications for diagnosis and management of the condition. PMID- 12539028 TI - Regional odontodysplasia: expression of matrix metalloproteinases and their natural inhibitors. AB - Regional odontodysplasia is a localized disorder of tissues of dental origin that results in a ghost-like appearance of the affected teeth. We present a case with a study of gingival tissue around the follicle. The results show evidence of the role of the matrix metalloproteinases and their natural inhibitors by resident cells in this pathosis. An imbalance in the amounts of matrix metalloproteinases and their natural inhibitors is associated with the pathologic breakdown of the collagen. PMID- 12539029 TI - Intraoral benign mesenchymoma: a report of 10 cases and review of the literature. AB - A benign mesenchymoma is an unencapsulated soft tissue neoplasm composed of 2 or more mature mesenchymal tissues not normally associated with each other, excluding fibrous connective tissue. No single mesenchymal tissue should predominate with respect to the other mesenchymal elements. Ten well-documented examples of intraoral benign mesenchymoma have been reported in the English language literature. The purpose of this report is to document 10 additional cases and to review the clinicopathologic characteristics of this uncommon tumor. PMID- 12539030 TI - Lymphoid papillary hyperplasia: report of a case. AB - A rare case of papillary lymphoid hyperplasia is reported in a 16-year-old girl. The case presented as papillomatous lesions of both palatine tonsils. Histopathologic findings revealed an unusual form of lymphoid hyperplasia. It is important to recognize this peculiar lesion because it exhibits clinical features suggestive of benign (papillomas) and malignant (carcinoma) neoplasia. Papillary lymphoid hyperplasia is a benign process, probably non-neoplastic, and easily cured by tonsillectomy. PMID- 12539031 TI - Human papillomavirus, type 40-associated papilloma, and concurrent Kaposi's sarcoma involving the anterior hard palate of an HIV-positive man. AB - A number of oral lesions have been reported in association with HIV, including lesions caused by other viruses such as the epitheliotropic human papillomavirus (HPV). More than 90 types of HPV have been identified, with the less commonly encountered strains of HPV tending to show association with immunodeficiency states. In addition, HIV-infected patients may have Kaposi's sarcoma develop, a malignancy thought to be caused by human herpes virus, type 8. Recent evidence suggests a sexual mode of transmission for this virus. We report an HIV-positive man with a large, HPV type 40-associated papilloma of the anterior palate and a previously undiagnosed focus of Kaposi's sarcoma. PMID- 12539032 TI - Significance of histopathologic examination in the diagnosis of dentin defects associated with type IV osteogenesis imperfecta: two case reports. AB - Two cases of type IV osteogenesis imperfecta, which is divided into subtypes A and B on the basis of the absence or the presence of dental alterations, respectively, are examined with respect to their dental features clinically, radiographically, and histopathologically. There were no characteristic dental abnormalities noted on the clinical and radiographic examination. However, the histopathologic examination with both light and electron microscopy disclosed characteristic dentin defects such as unevenly calcified matrixes, irregular tubular patterns, obliterated dentinal tubules, and cellular inclusions in the circumpulpal dentin of primary teeth. As a result, the patients were diagnosed as having osteogenesis imperfecta type IVB, although the clinical dental alterations were scarcely apparent. These findings indicate the importance of histopathologic examination with light and transmission electron microscopy for accurate diagnosis of osteogenesis imperfecta. PMID- 12539033 TI - Persistent acantholytic dermatosis of the lip related to solar damage. AB - We describe the case of a 70-year-old woman with persistent acantholytic dermatosis. The peculiarity of the case lies in the unusual localization of this dermatosis on the lower lip. A possible relationship with chronic solar damage is discussed. PMID- 12539034 TI - Clinical evaluation of the measuring accuracy of ROOT ZX in primary teeth. AB - OBJECTIVE: The aim of this study was to assess the accuracy of an electronic device (Root ZX; Morita, Tokyo, Japan) for measuring the root canal length in primary teeth. STUDY DESIGN: The pulp tissue was removed from 71 nonrestorable teeth scheduled to be extracted under general anesthesia, and the root canals (n = 105) were irrigated (H(2)O(2), 3%; NaOCl, 1%). Subsequently, the length was determined clinically with the electronic device prior to extraction. Treatments were performed by 2 dentists (42 and 63 root canals). After extraction, the real length was recorded and the 2 measurements were compared. RESULTS: Measurements were affected significantly by the dentists (P < .01; chi(2)). However, regression analysis revealed sufficient accuracy of the device, with a tendency to estimate the root canal length just short (x = -0.98 +/- 1.75 mm) of the apex. These results were not influenced by tooth type, root canal type, status of the periapex, or clinical condition (P > .05; chi(2)). CONCLUSION: Root ZX can be strongly recommended for clinical implementation of endodontics in primary teeth, particularly when treating fidgety children. PMID- 12539035 TI - The hemolytic and cytotoxic properties of a zeolite-containing root filling material in vitro. AB - OBJECTIVE: This in vitro study characterized the hemolysis and cytotoxicity of ZUT, an experimental glass ionomer cement (GIC) sealer with an added antimicrobial-containing zeolite (0.2% Zeomic w/w). STUDY DESIGN: ZUT, Ketac-Cem (GIC component of ZUT), Ketac-Endo, and two AH 26 sealer formulations were tested at various times after mixing. Hemolysis produced by standardized specimens was determined spectrophotometrically (n = 6/material). Cytotoxicity was assessed by using a Millipore Filter test with a HeLa cell monolayer (n = 10/material). Tests were repeated, and results were analyzed with a one-way analysis of variance (alpha = .05). RESULTS: Disks of AH 26 containing silver produced the most hemolysis of all test groups (P < .0001). Compared to controls, GICs and AH 26 formulations were noncytotoxic at 1 and 6 hours after mixing, respectively (P > .05). Addition of Zeomic did not increase the cytotoxic and hemolytic activity of Ketac-Cem (P > .05). CONCLUSION: Overall results suggest ZUT is less cytotoxic than AH 26 and possesses characteristics similar to the other GIC formulations tested. PMID- 12539036 TI - Diagnostic value of tuned-aperture computed tomography versus conventional dentoalveolar imaging in assessment of impacted teeth. AB - OBJECTIVE: This clinical study compares the diagnostic value of interactive 3 dimensional tuned-aperture computed tomography (TACT) with conventional dentoalveolar radiographic examinations of impacted maxillary incisors. STUDY DESIGN: TACT was compared variously with conventional intraoral, occlusal, rotational panoramic, and lateral cephalographic examinations. Sixteen dentists independently evaluated the clarity of anatomic structures of impacted anterior maxillary teeth and surrounding tissues. Estimation of confidence in clinical assessment of the patient was made together with a determination of the diagnostic potential of the studied modalities for altering treatment plans. RESULTS: TACT was found to significantly improve depiction of the buccal/palatal position of the impacted tooth and its relationship with adjacent teeth (P < .0001) irrespective of the availability of lateral cephalograms. Subjective assessments for clinical decision making approximated a 10% increase in confidence ratings with TACT. The added diagnostic value with TACT varied from case to case. CONCLUSIONS: The interactive 3-dimensional TACT display was perceived to be more informative for assessing impacted teeth and their relation to surrounding tissues than conventional methods in some cases but not in others. TACT altered treatment-option selection in some instances. PMID- 12539037 TI - The accuracy of dental panoramic tomographs in determining the root morphology of mandibular third molar teeth before surgery. AB - OBJECTIVE: The objective of this study was to assess the accuracy of dental panoramic tomograph (DPT) in the presurgical assessment of mandibular third molar teeth by correlating the radiologic interpretation with surgical findings. STUDY DESIGN: DPTs of 300 mandibular third molar teeth were assessed by 9 staff oral surgeons for root morphology and proximity to the inferior alveolar neurovascular bundle. Detailed records were made at surgery. RESULTS: The sensitivity and specificity for observation of root curvatures (+/-15%) were 29% and 94%, respectively. The sensitivity and specificity of determining an intimate relationship between the root and the neurovascular bundle were 66% and 74%, respectively. Kappa analysis demonstrated poor levels of agreement between radiologic interpretation and surgical findings when the number of roots was determined (0.191) and also when the roots were fused or separate (0.466). CONCLUSION: Using DPTs to assess mandibular third molar teeth before surgery gives poor diagnostic accuracy of anatomic form and structures. PMID- 12539038 TI - Carbon monoxide suppresses arteriosclerotic lesions associated with chronic graft rejection and with balloon injury. AB - Carbon monoxide (CO), one of the products of heme oxygenase action on heme, prevents arteriosclerotic lesions that occur following aorta transplantation; pre exposure to 250 parts per million of CO for 1 hour before injury suppresses stenosis after carotid balloon injury in rats as well as in mice. The protective effect of CO is associated with a profound inhibition of graft leukocyte infiltration/activation as well as with inhibition of smooth muscle cell proliferation. The anti-proliferative effect of CO in vitro requires the activation of guanylate cyclase, the generation of cGMP, the activation of p38 mitogen-activated protein kinases and the expression of the cell cycle inhibitor p21Cip1. These findings demonstrate a protective role for CO in vascular injury and support its use as a therapeutic agent. PMID- 12539039 TI - Autoimmune islet destruction in spontaneous type 1 diabetes is not beta-cell exclusive. AB - Pancreatic islets of Langerhans are enveloped by peri-islet Schwann cells (pSC), which express glial fibrillary acidic protein (GFAP) and S100beta. pSC autoreactive T- and B-cell responses arise in 3- to 4-week-old diabetes-prone non obese diabetic (NOD) mice, followed by progressive pSC destruction before detectable beta-cell death. Humans with probable prediabetes generate similar autoreactivities, and autoantibodies in islet-cell autoantibody (lCA) -positive sera co-localize to pSC. Moreover, GFAP-specific NOD T-cell lines transferred pathogenic peri-insulitis to NOD/severe combined immunodeficient (NOD/SCID) mice, and immunotherapy with GFAP or S100beta prevented diabetes. pSC survived in rat insulin promoter Iymphocytic choriomeningitis virus (rip-LCMV) glycoprotein/CD8+ T-cell receptor(gp) double-transgenic mice with virus-induced diabetes, suggesting that pSC death is not an obligate consequence of local inflammation and beta-cell destruction. However, pSC were deleted in spontaneously diabetic NOD mice carrying the CD8+/8.3 T-cell receptor transgene, a T cell receptor commonly expressed in earliest islet infiltrates. Autoimmune targeting of pancreatic nervous system tissue elements seems to be an integral, early part of natural type 1 diabetes. PMID- 12539040 TI - VEGF: a modifier of the del22q11 (DiGeorge) syndrome? AB - Hemizygous deletion of chromosome 22q11 (del22q11) causes thymic, parathyroid, craniofacial and life-threatening cardiovascular birth defects in 1 in 4,000 infants. The del22q11 syndrome is likely caused by haploinsufficiency of TBX1, but its variable expressivity indicates the involvement of additional modifiers. Here, we report that absence of the Vegf164 isoform caused birth defects in mice, reminiscent of those found in del22q11 patients. The close correlation of birth and vascular defects indicated that vascular dysgenesis may pathogenetically contribute to the birth defects. Vegf interacted with Tbx1, as Tbx1 expression was reduced in Vegf164-deficient embryos and knocked-down vegf levels enhanced the pharyngeal arch artery defects induced by tbx1 knockdown in zebrafish. Moreover, initial evidence suggested that a VEGF promoter haplotype was associated with an increased risk for cardiovascular birth defects in del22q11 individuals. These genetic data in mouse, fish and human indicate that VEGF is a modifier of cardiovascular birth defects in the del22q11 syndrome. PMID- 12539041 TI - Measuring the frequency of mouse and human cytotoxic T cells by the Lysispot assay: independent regulation of cytokine secretion and short-term killing. AB - Antigen-specific T cells demonstrate several potent effector functions during immune responses. Direct killing of infected cells is crucial for clearing viruses and other intracellular pathogens, but it has been difficult to measure the frequency of cytolytic cells. We have now developed a single-cell assay to measure the number of cytotoxic cells in a population, using a herpes simplex virus amplicon vector to express Escherichia coli beta-galactosidase in mouse or human target cells, and an Elispot to detect release of beta-galactosidase from killed target cells. This antigen-specific, perforin-dependent Lysispot assay has been combined with a cytokine Elispot in a two-color assay to confirm that cytotoxicity and interferon-gamma secretion are regulated independently. The simultaneous enumeration of cytokine-secreting and cytotoxic cells should be invaluable for ex vivo analysis of immune responses during infection and autoimmunity. PMID- 12539042 TI - HIV-1 Tat reprograms immature dendritic cells to express chemoattractants for activated T cells and macrophages. AB - Immature dendritic cells are among the first cells infected by retroviruses after mucosal exposure. We explored the effects of human immunodeficiency virus-1 (HIV 1) and its Tat transactivator on these primary antigen-presenting cells using DNA microarray analysis and functional assays. We found that HIV-1 infection or Tat expression induces interferon (IFN)-responsive gene expression in immature human dendritic cells without inducing maturation. Among the induced gene products are chemokines that recruit activated T cells and macrophages, the ultimate target cells for the virus. Dendritic cells in the lymph nodes of macaques infected with simian immunodeficiency virus (SIV) have elevated levels of monocyte chemoattractant protein 2 (MCP-2), demonstrating that chemokine induction also occurs during retroviral infection in vivo. These results show that HIV-1 Tat reprograms host dendritic cell gene expression to facilitate expansion of HIV-1 infection. PMID- 12539043 TI - TICAM-1, an adaptor molecule that participates in Toll-like receptor 3-mediated interferon-beta induction. AB - Human Toll-like receptor (TLR) 3 recognizes double-stranded (ds) RNA and induces production of interferon (IFN)-beta independent of the adaptor molecules MyD88 and TIRAP. Thus, another adaptor must exist that preferentially mediates TLR3 dependent production of IFN-beta. We have identified an alternative adaptor, designated Toll-interleukin 1 receptor domain (TIR)-containing adaptor molecule (TICAM)-1, that can physically bind the TIR domain of TLR3 and activate the IFN beta promoter in response to poly(I):poly(C). Thus, dsRNA-TLR3-dependent production of IFN-beta is mediated mainly by TICAM-1. This TICAM-1-dependent pathway may have a role in other TLR-IFN-beta pathways, which form part of the MyD88-independent cellular immune response. PMID- 12539044 TI - Nuclear genetic control of mitochondrial DNA segregation. AB - Mammalian mitochondrial DNA (mtDNA) is a high copy-number, maternally inherited genome that codes for a small number of essential proteins involved in oxidative phosphorylation. Mutations in mtDNA are responsible for a broad spectrum of clinical disorders. The segregation pattern of pathogenic mtDNA mutants is an important determinant of the nature and severity of mitochondrial disease, but it varies with the specific mutation, cell type and nuclear background and generally does not correlate well with mitochondrial dysfunction. To identify nuclear genes that modify the segregation behavior of mtDNA, we used a heteroplasmic mouse model derived from two inbred strains (BALB/c and NZB; ref. 12), in which we had previously demonstrated tissue-specific and age-dependent directional selection for different mtDNA genotypes in the same mouse. Here we show that this phenotype segregates in F2 mice from a genetic cross (BALB/c x CAST/Ei) and that it maps to at least three quantitative-trait loci (QTLs). Genome-wide scans showed linkage of the trait to loci on Chromosomes 2, 5 and 6, accounting for 16-35% of the variance in the trait, depending on the tissue and age of the mouse. This is the first genetic evidence for nuclear control of mammalian mtDNA segregation. PMID- 12539045 TI - Heterozygous disruption of Hic1 predisposes mice to a gender-dependent spectrum of malignant tumors. AB - The gene hypermethylated in cancer-1 (HIC1) encodes a zinc-finger transcription factor that belongs to a group of proteins known as the POZ family. HIC1 is hypermethylated and transcriptionally silent in several types of human cancer. Homozygous disruption of Hic1 impairs development and results in embryonic and perinatal lethality in mice. Here we show that mice disrupted in the germ line for only one allele of Hic1 develop many different spontaneous malignant tumors, including a predominance of epithelial cancers in males and lymphomas and sarcomas in females. The complete loss of Hic1 function in the heterozygous mice seems to involve dense methylation of the promoter of the remaining wild-type allele. We conclude that HIC1 is a candidate tumor-suppressor gene for which loss of function in both mouse and human cancers is associated only with epigenetic modifications. PMID- 12539046 TI - Zygote arrest 1 (Zar1) is a novel maternal-effect gene critical for the oocyte-to embryo transition. AB - The female gamete (the oocyte) serves the distinct purpose of transmitting the maternal genome and other maternal factors that are critical for post-ovulation events. Through the identification and characterization of oocyte-specific factors, we are beginning to appreciate the diverse functions of oocytes in ovarian folliculogenesis, fertilization and embryogenesis. To understand these processes further, we identified genes called zygote arrest 1 (Zar1 and ZAR1 in mouse and human, respectively) as novel oocyte-specific genes. These encode proteins of 361 amino acids and 424 amino acids, respectively, which share 59% amino-acid identity and an atypical plant homeo-domain (PHD) motif. Although Zar1 null (Zar1(-/-)) mice are viable and grossly normal, Zar1(-/-) females are infertile. Ovarian development and oogenesis through the early stages of fertilization are evidently unimpaired, but most embryos from Zar1(-/-) females arrest at the one-cell stage. Distinct pronuclei form and DNA replication initiates, but the maternal and paternal genomes remain separate in arrested zygotes. Fewer than 20% of the embryos derived from Zar1(-/-) females progress to the two-cell stage and show marked reduction in the synthesis of the transcription-requiring complex, and no embryos develop to the four-cell stage. Thus, Zar1 is the first identified oocyte-specific maternal-effect gene that functions at the oocyte-to-embryo transition and, as such, offers new insights into the initiation of embryonic development and fertility control in mammals. PMID- 12539047 TI - Haploinsufficiency of ATP1A2 encoding the Na+/K+ pump alpha2 subunit associated with familial hemiplegic migraine type 2. AB - Headache attacks and autonomic dysfunctions characterize migraine, a very common, disabling disorder with a prevalence of 12% in the general population of Western countries. About 20% of individuals affected with migraine experience aura, a visual or sensory-motor neurological dysfunction that usually precedes or accompanies the headache. Although the mode of transmission is controversial, population-based and twin studies have implicated genetic factors, especially in migraine with aura. Familial hemiplegic migraine is a hereditary form of migraine characterized by aura and some hemiparesis. Here we show that mutations in the gene ATP1A2 that encodes the alpha2 subunit of the Na+/K+ pump are associated with familial hemiplegic migraine type 2 (FHM2) linked to chromosome 1q23 (OMIM 602481). Functional data indicate that the putative pathogenetic mechanism is triggered by a loss of function of a single allele of ATP1A2. This is the first report associating mutations of Na+K+ pump subunits to genetic diseases. PMID- 12539048 TI - Non-polarized targeting of AE1 causes autosomal dominant distal renal tubular acidosis. AB - Autosomal dominant distal renal tubular acidosis (ddRTA) is caused by mutations in SLC4A1, which encodes the polytopic chloride-bicarbonate exchanger AE1 that is normally expressed at the basolateral surface of alpha-intercalated cells in the distal nephron. Here we report that, in contrast with many disorders in which mutant membrane proteins are retained intracellularly and degraded, ddRTA can result from aberrant targeting of AE1 to the apical surface. PMID- 12539049 TI - t(11;19)(q21;p13) translocation in mucoepidermoid carcinoma creates a novel fusion product that disrupts a Notch signaling pathway. AB - Truncation of Notch1 has been shown to cause a subtype of acute leukemia, and activation of Notch4 has been associated with mammary and salivary gland carcinomas of mice. Here we identify a new mechanism for disrupting Notch signaling in human tumorigenesis, characterized by altered function of a new ortholog of the Drosophila melanogaster Notch co-activator molecule Mastermind. We cloned the t(11;19) translocation that underlies the most common type of human malignant salivary gland tumor. This rearrangement fuses exon 1 from a novel gene of unknown function at 19p13, termed mucoepidermoid carcinoma translocated 1 (MECT1), with exons 2-5 of a novel member of the Mastermind-like gene family (MAML2) at 11q21 (ref. 3). Similar to D. melanogaster Mastermind and MAML1 (refs. 4,5), full-length MAML2 functioned as a CSL (CBF-1, suppressor of hairless and Lag-1)-dependent transcriptional co-activator for ligand-stimulated Notch. In contrast, MECT1-MAML2 activated transcription of the Notch target gene HES1 independently of both Notch ligand and CSL binding sites. MECT1-MAML2 induced foci formation in RK3E epithelial cells, confirming a biological effect for the fusion product. These data suggest a new mechanism to disrupt the function of a Notch co-activator in a common type of malignant salivary gland tumor. PMID- 12539050 TI - Extensive allelic variation and ultrashort telomeres in senescent human cells. AB - By imposing a limit on the proliferative lifespan of most somatic cells, telomere erosion represents an innate mechanism for tumor suppression and may contribute to age-related disease. A detailed understanding of the pathways that link shortened telomeres to replicative senescence has been severely hindered by the inability of current methods to analyze telomere dynamics in detail. Here we describe single telomere length analysis (STELA), a PCR-based approach that accurately measures the full spectrum of telomere lengths from individual chromosomes. STELA analysis of human XpYp telomeres in fibroblasts identifies several features of telomere biology. We observe bimodal distributions of telomeres in normal fibroblasts; these distributions result from inter-allelic differences of up to 6.5 kb, indicating that unexpectedly large-scale differences in zygotic telomere length are maintained throughout development. Most telomeres shorten in a gradual fashion consistent with simple losses through end replication, and the rates of erosion are independent of allele size. Superimposed on this are occasional, more substantial changes in length, which may be the consequence of additional mutational mechanisms. Notably, some alleles show almost complete loss of TTAGGG repeats at senescence. PMID- 12539054 TI - Finding language in the matter of the brain: origins of the clinical aphasia examination. AB - The origins of the aphasia examination can be traced back to the 19th century when physicians and scientists began to understand how higher mental functions such as language could be localized in the brain. Paul Broca, Carl Wernicke, and Hughlings Jackson developed different models of brain function, and each contributed important insights to the study of aphasia. Broca's contributions were influenced by the fundamental question of whether higher mental function could be localized in the brain at all; Wernicke's contributions were influenced by an attempt to unite more mechanistic and physiological principles to a model of higher brain functions; and Jackson's contributions were influenced by British association psychology. In addition to reviewing the origins of the aphasia examination, this article reviews the historical context in which these contributors worked, the factors that affected the reception of their views, and the manner in which their views have affected the aphasia examination and understanding of aphasia today. PMID- 12539055 TI - Scientific neurology and the history of the clinical examination of selected motor cranial nerves. AB - The content of the neurological examination of the motor cranial nerves is a reflection of the current understanding of the neuroanatomy and clinical neurophysiology of the brain stem. The history of the neuroanatomy of the cranial nerves extends from Galen's investigations c. 200 CE to the determination of the innervations of the facial nerve by Charles Bell in the early 19th century. Scientific clinical neurophysiology first appears in the work of John Hughlings Jackson in the mid- to late 19th century. The modern model of the organization and function of the nervous system results in a clinical examination that can be called neurological. The codification of the technique of the neurological examination of the cranial nerves mirrors the institutional and social history of neurology and is reflected in the changing role of neurology textbooks in the 19th and 20th centuries. The texts develop through stages of classification, pedagogy, reference, and institutionalization and illustrate the development of a specialized medical discipline. PMID- 12539056 TI - The historical roots of the visual examination. AB - The history of the visual examination is discussed in five parts. The history of visual acuity is followed from the minimum separable of Persian scientists to the 19th-century charts. Events in the history of the examination of the pupil include the late discovery of the significance of anisocoria in trauma, the description of the pupillary light reflex, the midbrain nucleus responsible for it, and the discovery of the neuroanatomic basis of pupillary abnormalities. Attempts to look into the eyeball date from the 17th century and are followed up to the discovery of the ophthalmoscope (1851). The evolution of the visual field examination is described from the first observations of hemianopia in the Hippocratic Corpus, to the present day visual field examination. Important landmarks in the history of central disorders of visual integration include "mind blindness" and the theories on aphasia that formed the basis for the study of disorders of central integration. PMID- 12539057 TI - Motor testing in neurology: an historical overview. AB - Systematic testing of muscle tone, bulk, and strength and analysis of the distribution of abnormal findings is a key element of bedside cerebral localization. A codified neurological examination was developed in the 20th century, but physicians have commented on weakness since antiquity. The ancient Egyptians, Greeks, and Romans described weakness in patients with head injuries and strokes, explaining it in terms of the pathophysiology of their times. Diagnostic principles of lateralization and localization developed in the Enlightenment, alongside an understanding of the anatomy of the motor system. In the 19th century, the work of Romberg, Todd, Charcot, and Gowers popularized the use of motor localization by demonstrating the power of scientific reasoning at the bedside. The concept of the upper motor neuron, as elaborated by Gowers, has enduring clinical utility. PMID- 12539058 TI - The history of the development of the cerebellar examination. AB - The cerebellar examination evolved from observations of experimental lesions made by neurophysiologists and clinical descriptions of patients with trauma to the cerebellum. At the beginning of the 19th century, neurophysiologists such as Luigi Rolando, Marie-Jean-Pierre Flourens, and John Call Dalton, Jr. ablated portions of the cerebellum of a variety of animals and observed staggering gait, clumsiness, and falling from side to side without loss of strength. They concluded that the cerebellum coordinated voluntary movements. In 1899, Joseph Francois Felix Babinski observed that patients with cerebellar lesions could not execute complex movements without breaking down into their elemental movements and described the defect as dysmetria. In 1902, Babinski coined the term dysdiodochokinesis to describe the inability to perform rapid execution of movements requiring alternate contractions of agonist and antagonist muscles. Gordon Holmes in 1904 described the phenomena of rebound, noting that if a limb ipsilateral to a cerebellar lesion is suddenly released from tension, the appendage will flail. In 1917, Gordon Holmes reported hypotonia and dysmetria in men wounded by gunshot wounds to their cerebellum. These observations were rapidly included in descriptions of the cerebellar examination in popular contemporaneous textbooks of neurology. Modern observations have demonstrated that the cerebellum influences such cognitive functions such as planning, verbal fluency, abstract reasoning, prosody, and use of correct grammar. PMID- 12539059 TI - Erb and Westphal: simultaneous discovery of the deep tendon reflexes. AB - Although the deep tendon reflexes (DTRs) were first introduced simultaneously into the medical literature by Erb and Westphal (1875), there is some evidence that the knee jerk response was well known to laymen prior to that time. Erb wrote that while he was not calling attention to anything new, he realized the utility of the phenomenon in diagnosing certain disease states. Erb viewed the phenomenon more clearly as a true reflex arc, whereas Westphal interpreted it as a local muscle phenomenon. By 1885, the maneuver was used in Europe and across the Atlantic in America. By then, the DTRs had become one of the cardinal features of the neurological examination. PMID- 12539060 TI - History of the extensor plantar response: Babinski and Chaddock signs. AB - The testing of reflexes involving the lower extremities is a pivotal part of the modern neurological examination. The normal response to noxious stimulation of the foot is plantar flexion of the toes, causing them to curl downward toward the undersurface of the foot. In 1896, Joseph Babinski described an extensor toe response that he claimed was a consistent finding among patients with pyramidal tract lesions of the cortex, subcortex, brain stem, or spinal cord. He considered it a distinct sign of organic disease and found it to be absent in cases of hysterical weakness. Charles Gilbert Chaddock admired the work of Babinski and described a modification of the Babinski technique, demonstrating that stimulation of the lateral surface of the foot could induce the same type of toe extension in patients with pyramidal tract lesions. The two reflexes are complementary, and each can occur without the other, although both are usually present in cases of pyramidal tract impairment. Although these two reflexes are the most commonly described pathological reflexes indicative of pyramidal tract disruption, the names of other celebrated neurologists are also linked to the study of extensor toe signs, each having identified a variant of the Babinski sign. PMID- 12539061 TI - Origins of the sensory examination in neurology. AB - Formal testing of sensation as part of the neurological examination followed the improvements in examination techniques as well as advances in neuroscience. By the 1890s, the observation that temperature sense was frequently impaired at the same time that pain was appreciated led to the supposition that the two paths traveled closely. Through the works of Brown-Sequard and Edinger the existence of a crossed afferent tract was verified. The distinction between two sensory pathways was clear by 1898, when van Gehuchten reported a case of syringomyelia and suggested that the pain and temperature fibers were carried anterolaterally and the position sense fibers carried posteriorly in the spinal cord. Many authors describing patients with tabes dorsalis suspected the posterior columns of the spinal cord played a key role in position sense. It is difficult to determine in the 19th century who first employed the use of movements of joints as a test for proprioceptive function; however, Bell in 1826 recognized what he termed a sixth sense, which later was characterized as proprioceptive function. Goldscheider went on to report the degrees of movement that were considered normal for each joint. Although vibratory sense had been described by Cardano and Ingrassia in the 16th century and tests had been developed by Rinne and Rumpf by the 19th century, it was not until 1903 that Rydel and Seiffer found that vibratory sense and proprioceptive sense were closely related and that both senses were carried in the posterior columns of the spinal cord. By 1955, the sensory examination included tests for light-touch, superficial pain, temperature, position sense, vibration, muscle (deep pain), and two-point discrimination. Tests for these sensibilities still remain in use. We will review the origins of the understanding of sensation, which ultimately led to the development of the sensory examination. We will highlight individuals who made important discoveries and observations, as well as review the history of each of the elements of the sensory examination. PMID- 12539062 TI - The Romberg sign and early instruments for measuring postural sway. AB - In the first half of the 19th century, European physicians-including Marshall Hall, Moritz Romberg, and Bernardus Brach-described loss of postural control in darkness of patients with severely compromised proprioception. Romberg and Brach emphasized the relationship between this sign and tabes dorsalis. Later, other neurologists evaluated the phenomenon, which is now known as Romberg's sign, in a broader range of neurologic disorders using a variety of simple but increasingly precise and sensitive clinical tests. In the late 19th century, neurologists also developed instruments to measure and record postural sway in patients with neurologic disease. Principal contributors included Philadelphia neurologist Silas Weir Mitchell and his trainees Morris Lewis and Guy Hinsdale. The efforts of these neurologists anticipated later physiologic studies and ultimately the development of computerized dynamic platform posturography. PMID- 12539063 TI - Epidemic Rift Valley fever in Saudi Arabia: a clinical study of severe illness in humans. AB - We describe the clinical patterns and case-fatality rate associated with severe Rift Valley fever (RVF) in patients who were admitted to the Gizan regional referral hospital during an outbreak of RVF in Saudi Arabia from September through November 2000. A total of 165 consecutive patients (136 men and 29 women) were prospectively studied; all were identified according to a strict case definition, were confirmed to have RVF by serologic testing, and were treated according to a predetermined protocol. The major clinical characteristics of RVF included a high frequency of hepatocellular failure in 124 patients (75.2%), acute renal failure in 68 patients (41.2%), and hemorrhagic manifestations in 32 patients (19.4%). Sixteen patients had retinitis and 7 patients had meningoencephalitis as late complications in the course of the disease. A total of 56 patients (33.9%) died. Hepatorenal failure, shock, and severe anemia were major factors associated with patient death. PMID- 12539064 TI - Development of a predictive index for picornavirus infections. AB - To create a clinical prediction index that aids in the diagnosis of picornavirus respiratory infections, we analyzed patients from 5 clinical trials designed to evaluate the efficacy of an antiviral treatment for respiratory infections. Logistic regression was used to determine which baseline symptoms and patient characteristics best predicted picornavirus infection. Parameter estimates were then used to create a predictive index for estimating the probability of picornavirus infection on the basis of cold symptoms. The presence at baseline of rhinorrhea (odds ratio [OR], 2.73), nasal congestion (OR, 1.63), and sore throat (OR, 1.37) increased the likelihood of picornavirus infection; the presence of myalgia (OR, 0.71) and fever (OR, 0.59) decreased the likelihood. The positive and negative predictive values of the model were 61.5% and 64.4%, respectively. The model was simplified for clinical use by creating a whole-number index: the lowest possible score (-3) indicates a 15% chance of picornavirus infection and the highest (7) indicates a 69% chance of picornavirus infection. PMID- 12539065 TI - A pharmacoeconomic evaluation of 7-valent pneumococcal conjugate vaccine in Canada. AB - The objective of this study was to evaluate the projected health benefits, costs, and cost-effectiveness of pneumococcal conjugate vaccination for infants and children aged <5 years in Canada. A health state model incorporating incidence, vaccine efficacy, costs, and transitional probabilities for the health states (well, meningitis, bacteremia, otitis media, pneumonia, and death) was constructed for a 10-year time horizon. Implementation of a pneumococcal conjugate vaccine program in Canada for each annual birth cohort of 340,000 persons observed over 10 years would be expected to save approximately 12 lives and 100,000 cases of pneumococcal disease over 10 years, resulting in total savings of $67 million (Canadian dollars [Can$]). Vaccination of healthy infants would result in net savings for society if the vaccine costs less than Can$50 per dose. Moreover, for a vaccine purchase price of Can$67.50, infant vaccination would cost society Can$79,000 per life-year gained. Pneumococcal conjugate vaccination is a potentially cost-effective means of pneumococcal disease prevention. PMID- 12539066 TI - Proinflammatory cytokine reactions in enterovirus 71 infections of the central nervous system. AB - Enterovirus 71 (EV71) infection can lead to devastating clinical outcomes. An appreciation of the scientific relationship between cytokine response and patient mortality may help limit the risks posed by this deadly illness. We present the results of a study that compared the cerebrospinal fluid (CSF) and serum levels of interleukin-6 (IL-6) and interleukin-1beta (IL-1beta) in 24 patients with EV71 infection. Cases in this study involved diverse manifestations or complications, including encephalitis, poliomyelitis-like syndrome, meningitis, and pulmonary edema. CSF levels of IL-6 in study patients were found to be consistently higher during the first 2 days of central nervous system (CNS) involvement than afterward. Compared with patients who did not have pulmonary edema, patients who experienced pulmonary edema had dramatically varied blood values, including IL-6, white blood cell counts, and glucose levels. Our findings suggest that the combination of CNS and systemic inflammatory response may trigger EV71-related cardiopulmonary collapse. PMID- 12539067 TI - Chronic hepatitis C virus infection in childhood: clinical patterns and evolution in 224 white children. AB - The characteristics and evolution of hepatitis C virus (HCV) infection were retrospectively investigated in a study of 224 HCV RNA-seropositive white children who were consecutively recruited at 7 European centers in 1980-1998. At presentation, all patients were positive for antibodies to hepatitis C virus, 87% were asymptomatic, and 48% had alanine aminotransferase (ALT) levels that were < or =2 times the upper limit of the range considered to be normal. Of 200 children followed for 1-17.5 years (mean follow-up +/- standard deviation [SD], 6.2+/-4.7 years), only 12 (6%) achieved sustained viremia clearance and normalization of the ALT level. In 92 revised liver biopsy specimen analyses, the mean fibrosis score (+/-SD) was 1.5+/-1.3 for children <15 years of age and 2.3+/-1.2 for children > or =15 years of age (range, 0-6 years; P<.01). Pediatric HCV infection is usually mild, but few patients, especially those who are perinatally infected, clear viremia in the medium-term follow-up. Conversely, the higher rates of fibrosis observed in older patients suggest the possibility of an insidious progression of HCV-associated liver disease. PMID- 12539068 TI - Risk of methicillin-resistant Staphylococcus aureus infection after previous infection or colonization. AB - Studies evaluating the risk of methicillin-resistant Staphylococcus aureus (MRSA) associated sequelae in colonized or infected inpatients have not extended follow up into the period after discharge from the hospital. We determined the 18-month risk of MRSA infection among 209 adult patients newly identified as harboring MRSA. Twenty-nine percent of patients (60 patients) developed subsequent MRSA infections (90 infections). These infections were often severe. Twenty-eight percent of infections (25 of 90) involved bacteremia, and 56% (50 of 90) involved pneumonia, soft tissue infection, osteomyelitis, or septic arthritis. Eighty percent of patients (48 of 60) with subsequent MRSA infection developed the infection at a new site, and 49% of new MRSA infections (44 of 90) first became manifest after discharge from the hospital. Accurate assessment of the risk of MRSA-associated sequelae requires prolonged follow-up after discharge. PMID- 12539069 TI - Evaluation of the immunochromatographic Binax NOW assay for detection of Streptococcus pneumoniae urinary antigen in a prospective study of community acquired pneumonia in Spain. AB - We evaluated the Binax NOW rapid immunochromatographic membrane test (ICT) for detection of Streptococcus pneumoniae urinary antigen in a population-based prospective study of adults with community-acquired pneumonia (CAP). ICT was performed with urine samples obtained from 452 (91.7%) of 493 patients enrolled. Pneumococcal antigen was detected in 19 (70.4%) of 27 patients with pneumococcal pneumonia. The test results were more frequently positive for patients who had not received antibiotics before testing (26.6% vs. 12.1%; P=.002). Only 16 (10.3%) of 156 samples obtained from patients with nonpneumococcal pneumonia yielded a positive result. Of the 269 patients who had pneumonia with no pathogen identified, antigen was detected in 69 (25.7%). With conventional microbiological criteria used as the "gold standard," the test had a sensitivity of 70.4% and a specificity of 89.7%. Testing concentrated urine samples with the ICT may be a useful technique for rapid diagnosis of pneumococcal pneumonia in adults with CAP. PMID- 12539070 TI - Isoniazid hepatotoxicity among drug users: the role of hepatitis C. AB - The incidence of and risk factors associated with hepatotoxicity in patients with chronic hepatitis have not been systematically studied. Therefore, we conducted a prospective study that included former drug users who were treated with isoniazid for latent tuberculosis infection. Of 415 patients, 20 (4.8%; 95% confidence interval [CI], 3-7.4) had hepatotoxicity diagnosed, and 6 (1.4%; 95% CI, 0.5-3.2) developed clinical hepatitis, none of whom had serious symptoms. The only 2 factors independently associated with isoniazid hepatotoxicity were excessive alcohol consumption (odds ratio [OR]; 4.2, 95% CI, 1.6-10.8; P=.002) and a high baseline alanine transaminase level (OR, 4.3; 95% CI, 1.6-11.4; P=.002). The presence of hepatitis C virus antibodies was associated with hepatotoxicity only on univariate analysis. Treatment with isoniazid in drug users appears to be safe and well tolerated, although frequent asymptomatic elevations in transaminase levels were observed. PMID- 12539071 TI - Influenza A and B virus infections in children. AB - To obtain data on the clinical manifestations of infection, the age distribution, and the underlying conditions of children with influenza severe enough to lead to hospital referral, we performed a retrospective study of children treated at Turku University Hospital (Turku, Finland) in 1980-1999. Influenza A or B antigen was detected in the nasopharyngeal aspirates of 683 of the 15,420 children studied. The median age of children with influenza A was 2.0 years (n=544), and that of children with influenza B was 4.2 years (n=139) (P<.001). One-fourth of the children had an underlying medical condition. High fever, cough, and rhinorrhea were the most frequently recorded symptoms. Acute otitis media developed in 24% of the children, and pneumonia developed in 9% of the children. The study shows that the majority of patient hospitalizations for pediatric influenza involve previously healthy infants and young children. Laboratory confirmation of influenza is particularly important for children because the clinical presentation of the infection is less characteristic than that seen in adults. PMID- 12539072 TI - Hemophagocytic syndrome in children: an important diagnostic consideration in fever of unknown origin. AB - To study the evolution of hemophagocytic syndrome (HPS) in children, we performed a retrospective review of 19 patients (median age, 17.4 months) in whom an infectious diseases consultation was requested at Texas Children's Hospital during the period of September 1991 through September 2001. Clinical findings consistent with HPS most frequently presented during days 6-14 of illness, concomitant with laboratory abnormalities. Fever was present for a median of 19 days before the diagnosis of HPS. Elevated serum lactate dehydrogenase and ferritin levels were noted in all patients. An infectious agent was identified in 42% of patients; 16% were found to have immunologic or vasculitic disease. HPS is a rare but often fatal disease that can initially present as fever of unknown origin with varying clinical findings, and it can be recognized by physicians who are familiar with the evolution of HPS. It is likely that many of these cases remain undiagnosed because of the HPS's rapidly fatal course. PMID- 12539073 TI - Clinical experience with linezolid for the treatment of nocardia infection. AB - Linezolid is an oxazolidinone that has activity against most gram-positive bacteria, including in vitro activity against all Nocardia species and strains. We describe 6 clinical cases of nocardiosis that were successfully treated with linezolid. Two patients had underlying X-linked chronic granulomatous disease, and 2 patients were receiving chronic corticosteroid therapy. Four of 6 patients had disseminated disease, and 2 of these 4 patients had multiple brain abscesses. Four patients primarily received monotherapy; for the fifth patient, linezolid was added to a failing multiple-drug regimen, and, for the sixth patient, it was used as part of combination therapy. All 6 patients were successfully treated, although 1 patient had a presumed relapse of central nervous system infection after premature discontinuation of the drug. Linezolid appears to be an effective alternative for the treatment of nocardiosis. PMID- 12539074 TI - Pneumococcal septic arthritis: review of 190 cases. AB - This article reports 13 cases of pneumococcal septic arthritis and reviews another 177 cases reported since 1965. Of 2407 cases of septic arthritis from large series, 156 (6%) were caused by Streptococcus pneumoniae. Mortality was 19% among adults and 0% among children. Pneumococcal bacteremia was the strongest predictor of mortality. At least 1 knee was involved in 56% of adults. Polyarticular disease (36%) and bacteremia (72%) were more common among adults with septic arthritis caused by S. pneumoniae than among adults with other causative organisms. Only 50% of adults with pneumococcal septic arthritis had another focus of pneumococcal infection, such as pneumonia. Functional outcomes were good in 95% of patients. Uncomplicated pneumococcal septic arthritis can be managed with arthrocentesis and 4 weeks of antibiotic therapy; most cases of pneumococcal prosthetic joint infection can be managed without prosthesis removal. A fatal case of septic arthritis caused by a beta-lactam-resistant strain of S. pneumoniae is also presented. PMID- 12539075 TI - Clinical features that discriminate inhalational anthrax from other acute respiratory illnesses. AB - Inhalational anthrax (IA) is a rapidly progressive disease that frequently results in sepsis and death, and prompt recognition is critical. To distinguish IA from other causes of acute respiratory illness, patients who had IA were compared with patients in an ambulatory clinic who had influenza-like illness (ILI) and with hospitalized patients who had community-acquired pneumonia (CAP) at the initial health care visit. Compared with patients who had ILI, patients who had IA were more likely to have tachycardia, high hematocrit, and low albumin and sodium levels and were less likely to have myalgias, headache, and nasal symptoms. Scoring systems were devised to compare IA with ILI or CAP on the basis of strength of association. For ILI, a score of > or =4 captured all 11 patients with IA and excluded 664 (96.1%) of 691 patients with ILI. Compared with patients who had CAP, patients with IA were more likely to have nausea or vomiting, tachycardia, high transaminase levels, low sodium levels, and normal white blood cell counts. For CAP, a score of > or =3 captured 9 (81.8%) of 11 patients with IA and excluded 528 (81.2%) of 650 patients with CAP. In conclusion, selected clinical features of patients with IA differ from those of patients with ILI and are more similar to those of patients with CAP. PMID- 12539076 TI - Primary cutaneous cryptococcosis: a distinct clinical entity. AB - Cryptococcus neoformans is an encapsulated yeast responsible for disseminated meningitis in immunocompromised hosts. Controversies persist on the existence of primary cutaneous cryptococcosis (PCC) versus cutaneous cryptococcosis being only secondary to hematogenous dissemination. Thus, we reviewed cryptococcosis cases associated with skin lesions reported in the French National Registry. Patients with PCC (n=28) differed significantly from those with secondary cutaneous cryptococcosis (n=80) or other forms of the disease (n=1866) by living area (mostly rural), age (older), ratio of men to women (approximately 1:1), and the lack of underlying disease. Evidence of PCC included the absence of dissemination and, predominantly, a solitary skin lesion on unclothed areas presenting as a whitlow or phlegmon, a history of skin injury, participation in outdoor activities, or exposure to bird droppings, and isolation of C. neoformans serotype D. Therefore, PCC is a distinct epidemiological and clinical entity with a favorable prognosis even for immunocompromised hosts. PMID- 12539077 TI - Clinical significance of Helicobacter species other than Helicobacter pylori. AB - The cultivation of Helicobacter pylori and the recognition of its clinical significance have served to stimulate interest in bacteria associated with the gastrointestinal and hepatobiliary tracts. Many novel Helicobacter species have been identified and are increasingly recognized in association with human disease, most of which is likely acquired as a zoonosis. Because their identification can be difficult by use of routine methods available in the clinical laboratory, awareness of methods for diagnosis and treatment of these Helicobacter species is important, particularly in the evaluation of immunocompromised patients. PMID- 12539078 TI - Vaccine and immunization resources on the World Wide Web. AB - Abundant information regarding immunizations and vaccines is available on the World Wide Web. This report was prepared as a guide to reliable Internet sources for health care providers, researchers, and patients or parents interested in gaining Web-based information on these topics. I describe comprehensive Web sites about immunizations, meetings on immunizations, and clinical trials on vaccines and for journals and publications on vaccines, organizations providing immunization information, and pharmaceutical companies and vaccine manufacturers. PMID- 12539079 TI - Influence of coinfection with hepatitis C virus on morbidity and mortality due to human immunodeficiency virus infection in the era of highly active antiretroviral therapy. AB - To ascertain the impact of hepatitis C virus (HCV) infection on human immunodeficiency virus (HIV) disease progression and associated death in the era of highly active antiretroviral therapy (HAART), we examined mortality rates, the presence of other diseases, and antiretroviral use in an observational cohort of 823 HIV-infected patients with and without HCV coinfection during the period of January 1996 through June 2001. Analyses were used to compare patient characteristics, comorbid conditions, and survival durations in HIV-infected and HIV-HCV-coinfected patients. HIV-HCV-coinfected persons did not have a statistically greater rate of acquired immunodeficiency syndrome or of renal or cardiovascular disease, but they did have more cases of cirrhosis and transaminase elevations. There were proportionately more deaths in the HIV-HCV coinfected group. Age, baseline CD4+ cell count, and duration of HAART were significantly associated with survival, but HCV infection was not. HAART use was a strong predictor of increased duration of survival, suggesting that treatment is more important to survival than is HCV coinfection status. PMID- 12539080 TI - First things first: balancing hepatitis C and human immunodeficiency virus. PMID- 12539081 TI - Growth hormone receptor (GH)-expressing carcinoid tumors after recombinant human GH therapy for human immunodeficiency virus-related lipodystrophy. AB - We describe a patient who had growth hormone receptor-expressing carcinoid tumors develop in the distal colon and rectum after he received recombinant human growth hormone therapy for human immunodeficiency virus-related lipodystrophy. This case report serves as a cautionary note regarding the use of potentially oncogenic recombinant human growth hormone therapy to treat human immunodeficiency virus positive persons. PMID- 12539082 TI - Methods for integration of pharmacokinetic and phenotypic information in the treatment of infection with human immunodeficiency virus. AB - Interest in monitoring antiretroviral drug concentrations in human immunodeficiency virus-infected patients has gained considerable momentum in recent years. We present a potential method for integrating pharmacokinetic and phenotypic information that will assist clinicians in choosing optimal treatment regimens for their patients and that will provide an approach for the interpretation of antiretroviral plasma drug concentrations. PMID- 12539083 TI - Fluoroquinolone resistance in Streptococcus pyogenes. AB - Streptococcus pyogenes isolated from blood and urine samples obtained from a 78 year-old woman was tested for susceptibility, and fluoroquinolone resistance (minimum inhibitory concentration of levofloxacin, 16 microg/mL) was found. DNA amplification and sequencing revealed a serine81-->tyrosine substitution in gyrA and 2 substitutions in parC: serine79-->phenylalanine and alanine121-->valine. This is the second report of a clinical isolate of S. pyogenes with high-level fluoroquinolone resistance. PMID- 12539084 TI - Optimal criteria for the diagnosis of Legionella coinfection. PMID- 12539085 TI - Nail pigmentation and fatigue in a 39-year-old woman. PMID- 12539086 TI - Human herpesvirus 8 variants in Venezuelan patients with AIDS-related Kaposi sarcoma. PMID- 12539087 TI - Bradycardic syncope in 2 patients who recently began gatifloxacin treatment. AB - Gatifloxacin is a synthetic broad-spectrum 8-methoxyfluoroquinolone approved by the United States Food and Drug Administration in December 1999. Few side effects of this new antibiotic have been reported, and there are no previous case reports of bradyarrhythmias. We report 2 cases of syncope due to bradycardia in patients who recently began treatment with gatifloxacin. PMID- 12539088 TI - Female-to-female transmission of human immunodeficiency virus. AB - We describe a case of female-to-female transmission of human immunodeficiency virus (HIV). A 20-year-old African American woman with no obvious risk factors received a diagnosis of HIV infection, and the genotype of the infecting strain closely matched that of the strain infecting her openly bisexual female partner. The route of transmission was probably use of sex toys, used vigorously enough to cause exchange of blood-tinged body fluids. PMID- 12539089 TI - Meningitis due to Capnocytophaga canimorsus after receipt of a dog bite: case report and review of the literature. AB - We describe a case of meningitis due to Capnocytophaga canimorsus and review 18 cases with attention to risk factors, clinical features, diagnosis, treatment, and outcome. In most of the reported cases, contact with dogs and predisposing factors were found. Clinical manifestations and the findings of examinations of cerebrospinal fluid specimens were similar to those of classic bacterial meningitis; however, the mortality rate for C. canimorsus meningitis very low when compared with the rate for C. canimorsus septicemia (5% vs. 30%). PMID- 12539090 TI - Enteroviral meningoencephalitis after anti-CD20 (rituximab) treatment. AB - Treatment with the chimeric anti-CD20 monoclonal antibody rituximab induces rapid and long-lasting depletion of circulating B cells. We report the occurrence of enteroviral meningoencephalitis following rituximab therapy in 1 child with immune thrombocytopenia and in 1 adult patient with relapsed B cell lymphoma. PMID- 12539091 TI - Acute cerebellitis caused by herpes simplex virus type 1. AB - Cerebellar disorders due to herpes simplex virus (HSV) infection are rare and always associated with herpes simplex encephalitis. We report 2 cases of severe primary acute cerebellitis caused by HSV type 1 that were identified by nested polymerase chain reaction performed on cerebrospinal fluid samples. PMID- 12539092 TI - Hot tub-associated necrotizing pneumonia due to Pseudomonas aeruginosa. AB - We describe a case of severe necrotizing pneumonia due to community-acquired Pseudomonas aeruginosa. Cultures of fluid obtained from the filter of the patient's hot tub grew the same P. aeruginosa strain as that grown from culture of the patient's sputum. Centers for Disease Control and Prevention guidelines should be strictly followed for hot tub maintenance to prevent P. aeruginosa overgrowth: the range of free chlorine levels in the water should be kept at 1-3 mg/L, and the pH should be kept at 7.2-7.8. PMID- 12539093 TI - Electrocardiographic potential correlations: rationale and basis for lead selection and ECG estimation. AB - Einthoven gave to us the electrocardiogram. Electrocardiographic mapping demonstrated that localized electrophysiological events and phenomena have localized body surface electrocardiographic manifestations. Clinical electrocardiography has given us "reasonably good" means (criteria) with which to detect and characterize medically significant cardiac conditions, events, and diseases. However, clinical electrocardiography is imperfect, largely as a consequence of inadequate or redundant spatial sampling. This compromises the sensitivity and specificity of diagnosing cardiac diseases for which electrocardiographic manifestations are present but undetected due to imperfect sampling that results in a low signal-to-noise ratio for the specific abnormality. Correlation structure of body-surface potential distributions between and across populations or individuals provides important insight into and justification for the selection and use of "limited", "reduced", or "derived" lead systems aimed at improving the capture and use of electrocardiographic information. In this paper, we show the electrocardiographic voltage correlation relationships that occur on the body surface, across groups of subjects, either with or without cardiac disease. In addition, we demonstrate the correlation relationships between torso-surface and epicardial-surface potential distributions in experiments incorporating isolated canine hearts in a human shaped torso tank. Analysis of these correlation relationships provides an explanation for the long-standing success of clinical electrocardiography but also suggests the means to improve its performance by incorporating new leads and/or their estimation from appropriately selected leads. PMID- 12539094 TI - How many electrodes and where? A "poldermodel" for electrocardiography. AB - It is known that body surface potential maps (BSPMs) contain diagnostic information not easily retrieved from the standard 12-lead electrocardiogram (ECG). Alternate lead sets that can reconstruct the BSPM have been proposed, but they are impractical because they require more than the 10 electrodes of the standard ECG and use nonstandard positions. We propose a practical approach to increasing the diagnostic information content of the standard ECG by repositioning selected chest electrodes. Thus, in the best tradition of the Dutch "poldermodel," we have sought to strike a compromise between the demands of different parties, one that, while suboptimal for each, will be seen by all as the best attainable result. We used a set of 746 120-lead BSPMs from healthy individuals and patients with various abnormalities. Data were split in a learning and a test set. Using the learning set, a general transformation to reconstruct all BSPM leads from the standard 12-lead ECG was derived by linear regression. Similarly, BSPMs were reconstructed when 2 of electrodes V(3)-V(6) were moved to other positions on the anterior part of the chest. Reconstruction performance was assessed on the test set by correlation and similarity coefficients. Thanks to the redundancy of information in the precordial leads, 2 missing precordial leads can be reconstructed from the others, using general coefficients, to a high degree of accuracy, particularly when the missing leads were not adjacent. We chose to reposition V(4) and V(6) to different sites on the anterior thorax. From the many electrode sites that we explored, those at 2 intercostal spaces below and above V(2) yielded better BSPM reconstruction than was attainable from the standard electrode positions, in most parts of the anterior thorax, including regions that are known to contain important diagnostic information less well brought forward by the standard ECG. Slight variations in the new electrode positions did not appreciably change the results. The standard 12-lead ECG proved to produce better overall reconstructions than either the EASI configuration or vectorcardiographic lead systems. Repositioning electrodes V(4) and V(6) provides a simple, practical method by which to improve the sampling of diagnostic information from the body surface while maintaining the full diagnostic content of the standard 12-lead ECG. This approach also obviates the need to determine the precise location of V(4) electrode, which may be difficult in women. PMID- 12539095 TI - Comparison of a new reduced lead set ECG with the standard ECG for diagnosing cardiac arrhythmias and myocardial ischemia. AB - In a few patients, 12-lead electrocardiograms (ECGs) derived from reduced-lead set configurations do not match the standard ECG. Constructing an ECG from a reduced number of standard leads should minimize this problem because some of the resultant 12 leads would always include "true" standard leads. The purpose of this study was to compare the ability of a new reduced-lead-set 12-lead ECG ("interpolated" ECG) with the standard ECG to diagnose cardiac arrhythmias and acute myocardial ischemia. The interpolated ECG uses six standard electrode sites (limb leads plus V(1) and V(5)), from which the remaining four precordial leads (V(2), V(3), V(4), and V(6)) are constructed. Standard and interpolated ECGs were compared using data from 2 prospective clinical trials involving 649 patients evaluated for 1) chest pain in the emergency department (ischemia group, n = 509) or 2) tachycardias in the cardiac electrophysiology laboratory (arrhythmia group, n = 140). Diagnoses were identical between standard and interpolated ECGs for bundle branch and fascicular blocks, left atrial enlargement, right ventricular hypertrophy, prior inferior myocardial infarction (MI), and the distinction of ventricular tachycardia from supraventricular tachycardia with aberrant conduction. There was 99% agreement for prior anterior MI (kappa, .935, P =.000). The percent agreement for acute myocardial ischemia on the initial ECG recorded in chest-pain patients in the emergency department was 99.2% (kappa, .978, P =.000). Of the 120 patients who had ST events with continuous standard 12-lead ECG monitoring, 116 (97%) also had criteria for transient ischemia with the interpolated ECG (ie, DeltaST >or= 100 microV in >or=1 lead(s) lasting >or=1 minute(s). The interpolated 12-lead ECG is comparable to the standard ECG for diagnosing multiple cardiac abnormalities, including wide-QRS-complex tachycardias and acute myocardial ischemia. The advantages of this ECG method are that the standard electrode sites are familiar to clinicians and that eight of the 12 leads are "true" standard leads. Hence, QRS-axis and morphology criteria for diagnosing wide-QRS-complex tachycardia and bundle branch and fascicular blocks are preserved. PMID- 12539096 TI - Improved EASI coefficients: their derivation, values, and performance. AB - The EASI lead system, which is based on the dipole hypothesis of vectorcardiography, offers the possibility of deriving the standard 12-lead electrocardiogram (ECG) and other desired leads from ECGs recorded at only 4 sites; it uses the Frank E, A, and I electrode locations, a fourth electrode location (S) at the manubrium, and a reference electrode. Accordingly, the electrodes of this system can be applied rapidly on easy-to-locate, stable anatomical sites that leave the precordium free for other diagnostic procedures. In early EASI implementations, the derived leads differed from actual leads by more than some clinicians found acceptable. As these differences were thought to be caused by the fact that the coefficients that were used had been derived from a limited data set, we have calculated a new set of EASI coefficients for the standard 12 leads, and several other leads, by using a data set of 983 adult subjects with 120-lead ECGs and well-documented diagnoses. This database is a concatenation of 2 previously described ones: one consisting of 892 persons (normal subjects, postmyocardial-infarction patients with and without arrhythmias, and patients with ventricular arrhythmias but no history of myocardial infarction) and the other consisting of 91 patients with single-vessel coronary artery disease who underwent coronary balloon-inflation angioplasty. In addition to the coefficients for the standard 12 leads (derived for standard limb leads as well as for Mason-Likar leads), we derived coefficients for six additional unipolar leads (posterior V(7)-V(9), and right-sided V(3)R-V(5)R), the Frank orthogonal leads, and three bipolar, vessel-specific leads that have been previously shown to exhibit optimal sensitivity for acute myocardial ischemia. We also derived coefficients for the modified electrode locations of the EASI system that must be used with patients who have undergone a midline sternotomy. Optimal coefficients for lead transformations were determined by maximizing the ensemble average (over the entire data set) of the correlation between the derived and the true lead for the chosen interval of the averaged complex. For derived standard limb leads, the amplitude was adjusted to give the best root-mean-square fit over the entire PQRST interval, whereas for derived Mason-Likar leads it was adjusted to give the best ST-segment fit. The entire set of coefficients and their corresponding goodness-of-fit measures are presented. PMID- 12539097 TI - Comparability of 12-lead ECGs derived from EASI leads with standard 12-lead ECGS in the classification of acute myocardial ischemia and old myocardial infarction. AB - We compared 12-lead electrocardiograms (ECGs) derived with an improved transformation matrix from EASI leads and standard 12-lead ECGs in the detection of acute myocardial ischemia and old infarction (MI). For the ischemia test, we used ECGs of 40 patients recorded prior to and at peak inflation during percutaneous transluminal coronary angioplasty, and for old MI we used test ECGs of 382 non-MI subjects and of 472 patients with prior MI documented by enzyme findings. Two experienced ECG readers served as separate, independent standards for lead-set comparisons, and the Philips ECG analysis program also classified the ECGs. The results showed no significant differences between the two lead sets in the detection of acute inflation-induced ischemia or of old MI according to coding by the electrocardiographers or the computer program. No significant differences were found between the electrocardiographers and the lead sets for acute ischemia. Classification differences between the electrocardiographers were larger than those between the lead sets for acute and old MI and were significant for the latter (P <.001). A more detailed comparison of the lead sets suggested a possible need for modified old-MI criteria and optimization of ST classification thresholds for acute ischemic injury, specific for the EASI 12-lead ECG. We conclude that the EASI-derived 12-lead ECG deserves serious consideration as an alternative to the standard 12-lead ECG in emergency situations and for monitoring in acute-care setting. PMID- 12539098 TI - Statistical and deterministic approaches to designing transformations of electrocardiographic leads. AB - Two different approaches can be used to investigate the relationships among electrocardiographic leads: a statistical one, based on the analysis of recorded electrocardiograms (ECGs), and a deterministic one, based on physical principles that govern the current flow in irregularly shaped volume conductors such as the human body. The purpose of this study was to compare these two approaches. For the statistical investigation, the data set consisted of 120-lead ECGs recorded in a population including normal subjects (n = 290), post-myocardial-infarction patients (n = 497), patients with a history of ventricular tachycardia but no evidence of a previous myocardial infarction (n = 105), and patients with a single-vessel coronary artery disease who underwent coronary angioplasty (n = 91). Lead transformations of interest were obtained by fitting the multiple regression model to this data set by the least-squares method. For the deterministic investigation, we used a boundary-element model of the human torso to simulate body-surface potentials in response to three orthogonal unit dipoles placed consecutively at 1,239 ventricular source locations, and the resulting body-surface potential distributions (instead of the recorded ECGs) were then fitted by the multiple-regression model. The results suggest that the lead transformations should be preferably designed by statistical analysis of recorded ECGs. Regression models with a small number of predictors (eg, those based on three ECG leads) are the most reliable; those using more predictors are fraught with the danger of collinearity when predictors are highly correlated (as occurs in the standard 12-lead ECG). Model-derived deterministic transformations are compatible with statistically derived ones, provided that the distributed character of the cardiac sources is taken into account. We conclude that statistical associations among electrocardiographic leads can be reliably quantified in sufficiently large and diverse databases of recorded data; the causality of these associations can be supported by appropriate deterministic models based on the laws of physics. PMID- 12539099 TI - Dobutamine stress magnetic resonance imaging: a valuable method in the noninvasive diagnosis of ischemic heart disease. AB - We assessed the clinical applicability of dobutamine stress magnetic resonance imaging (DS-MRI) for the detection of myocardial ischemia and myocardial viability. One hundred patients with suspected coronary artery disease and inconclusive exercise electrocardiography or significant repolarization abnormalities on the resting ECG underwent breath hold DS-MRI (1 Tesla), 4 days after cessation of anti-ischemic medication. Three left ventricular short axis planes were imaged at increasing doses of dobutamine. Recovery of wall thickening in a previously diminished or non contracting segment at low dose dobutamine was considered proof of viability. Development of hypo-, a- or dyskinesia at higher doses of dobutamine was taken to indicate ischemia. If the DS-MRI test was positive for ischemia, coronary angiography was performed. If indicated, this was followed by revascularization. If DS-MRI did not demonstrate ischemia, neither angiography nor revascularization were carried out. Ninety five DS-MRI investigations were available for diagnosis. Forty two patients had DS-MRI scans positive for ischemia and subsequently coronary angiography assessment of the clinical applicability of DS-MRI for the detection of myocardial ischemia was performed. One patient was false-positive. All 53 patients with non-ischemic DS MRI scans had follow-up for 11-23 months (mean 17 months). One patient died suddenly 2 weeks after the MRI-test. The other 52 patients did not experience any coronary event nor sudden cardiac death. The predictive value of a positive (for ischemia) DS-MRI test is 98% and the predictive value of a negative DS-MRI test is also 98%. PMID- 12539101 TI - Multielectrode venous catheter mapping as a high quality constraint for electrocardiographic inverse solution. AB - A persistent challenge in solving inverse problems in electrocardiography is the application of suitable constraints to the calculation of cardiac sources. Whether one formulates the inverse problem in terms of epicardial potentials or activation wavefronts, the problem is physically ill-posed and hence results in numerically unstable computations. Suitable physiological constraints applied with appropriate weighting can recover useful inverse solutions. However, it is often difficult to determine the best possible constraints and their optimal weighting. We have recently begun to use multielectrode catheters as a means of mapping epicardial signals in animal models. To accommodate the sparse sampling of this venous catheter based approach, we have applied statistical signal processing methods to estimate complete epicardial maps of activation time and epicardial potentials. Such measurements--and the estimated maps from them--also have the potential to provide high quality constraints for electrocardiographic inverse problems because they provide direct--albeit sparse--access to the desired solution. In this presentation we describe several approaches we have applied to extract useful constraints from sparsely sampled epicardial signals as well as a training set of epicardial maps, and use them to improve the quality of computed inverse solutions. Results suggest that combining various information sources provides valuable constraint information. Such a multimodal approach to cardiac mapping is clinically and technically viable and offers a possible means to overcome a major remaining limitation of inverse electrocardiography. PMID- 12539100 TI - Electromechanical mapping with MRI tagging and epicardial sock electrodes. AB - Methods currently exist for the precise measurement of local three-dimensional myocardial motion noninvasivly with magnetic resonace imaging tagging. From these motion estimates, strain images representing the local deformation of the myocardium can be formed to show local myocardial contraction. These images clearly show the sequence of mechanical events during the activation and relaxation of the heart, making them ideal to visualize abnormalities caused by asynchronous electrical activation or ischemia. Coupled with the near simultaneous mapping of electrical depolarization with a sock electrode array, we can investigate the relationship between electical activity and mechanical function on a local level in the in vivo heart. Registered fiber angle maps can be also be obtained in the same heart with diffusion magnetic resonance imaging to assist in the construction of the electromechanical model of the whole heart. PMID- 12539102 TI - Noninvasive determination of the activation sequence of the heart: application to patients with previous myocardial infarctions. AB - The uniform double layer (UDL) source model can be used to estimate the activation sequence of the heart at the ventricular surface from electrocardiogram (ECGs) measured on the body surface. Over the past decade, promising results have been obtained. However, the clinical value of the UDL model still has to be demonstrated. This paper reports on the results of experiments to validate the UDL-based method in patients who underwent open-chest surgery. The patients had previously suffered from ventricular arrhythmias after myocardial infarction. Prior to surgery, the ECG was recorded in 123 leads on the body surface at Helsinki University Central Hospital (HUCH). Based on MR images an individual volume conductor model was constructed for each patient. During surgery at HUCH, potentials were recorded invasively in 102 bipolar leads on the epicardium. Based on these data, activation maps were constructed. These maps were compared to the activation maps obtained by the UDL based inverse applied to the body-surface ECG data. The results showed that the overall pattern of the activation sequence found by the UDL corresponded well to the actual activation pattern for some patients. However, the results were poor for those patients whose heart had an infarcted region of considerable size. In a follow-up study a method was developed to take infarction into account in the UDL-based inverse procedure. The results of this study showed that one cannot expect good results from the UDL-based inverse in the presence of an old myocardial infarction unless that infarction is accounted for in the inverse procedure. PMID- 12539103 TI - Clinical ECG mapping and imaging of cardiac electrical excitation. AB - Combining electrocardiographic mapping and 3D+time anatomical data enables noninvasively the imaging of the electrical excitation sequence in the human heart. A bidomain-theory based surface heart model activation time imaging approach was employed to image single beat data of atrial and ventricular depolarisation. Activation time maps were reconstructed for three patients who underwent an electrophysiologic study. The sinus rhythm and a rhythm according to a pacing protocol were reconstructed for two patients. For the third patient the accessory pathway of the WPW syndrome was localized. For focal arrhythmias, this model-based imaging approach might allow the guidance and evaluation of antiarrhythmic interventions, for instance, in case of catheter ablation or drug therapy. PMID- 12539104 TI - Fiducial segment averaging to improve cardiac time interval estimates. AB - To improve the accuracy of fiducial point estimates, a technique was developed and tested that uses the coherence of relatively small segments within the P-QRS T complex from beat to beat. The methodology is applied to digitally stored electrocardiogram traces from various sources. It can be applied to single or multiple leads. First, the individual QRS complexes are identified with a matched filter technique. The trigger time points obtained are fine-adjusted by cross correlating in turn each individual complex with the average of the remainder complexes and then shifting the complex till maximum correlation is achieved. The process is repeated till no improvement can be attained. Then the individual fiducial points are identified and a similar fine-adjustment is made to the individual points with a segment of approximately 60 ms around the fiducial point. This process is performed in turn for P-onset, QRS-onset and offset, peak of T wave and end of T wave. The values of PR-interval, QRS-duration and QT follow from these measurements. The validity of the result can be assessed by visual inspection and by examining the standard deviation of the measurements obtained. From the above intervals, it is generally accepted that QRS-duration is fairly constant over a short recording duration, whereas PR and QT are inherently heart-rate dependent. Often a linear regression of QT onto RR can be calculated with a significant correlation coefficient, indicating a physiological QT/RR relationship in these recordings of relative short duration. It would be impossible to show this last relationship if the individual QT changes had not been preserved by the technique described. PMID- 12539105 TI - A software-based pacemaker pulse detection and paced rhythm classification algorithm. AB - A new pacemaker pulse detection and paced electrocardiogram (ECG) rhythm classification algorithm with high sensitivity and positive predictive value has been implemented as part of the Philips Medical Systems' (Andover, MA) ECG analysis program. The detection algorithm was developed on 1,108 paced ECGs with 16,029 individual pulse locations. It operates on 12-lead, 500 sample per second, 150 Hz low-pass filtered ECG signals. Even after low-pass filtering, this algorithm distinguishes between pacemaker pulses and narrow QRS complexes from newborns. An individual pulse detection sensitivity of 99.7% and positive predictive value of 99.5% was obtained by the multi-lead detector. A 10-second, 12-lead ECG database (n = 13,155) of paced (n = 2,190), non-paced adult (n = 8,070), non-paced pediatric (n = 1,209) and "noisy" ECGs with spike noise and muscle artifact (n = 1,686) was assembled and annotated by two readers. The overall performance in identification of an ECG as paced with any pacing present versus non-paced is 97.2% in sensitivity and 99.9% in specificity. The paced ECGs were classified by the mode in which the beats were paced, such as, atrial, ventricular, A-V dual, or dual/inhibited chamber (ie, combinations of atrial, ventricular and dual) pacing. An algorithm was developed for paced rhythm classification. The algorithm performance results show that accurate and robust pacemaker pulse detection and classification can be done in software on diagnostic bandwidth ECG signals. PMID- 12539106 TI - An improved automated ECG algorithm for detecting acute and prior myocardial infarction. AB - Commercial electrocardiographic (ECG) algorithms for detection of prior myocardial infarction (MI) predominantly rely on QRS criteria and on established qualitative ST and T changes. The qualitative approach to ST-T changes in Cardiovise 2.5 is improved upon in version 3.0 with two distinct new approaches for quantifying ST and T changes to assist with the detection of prior MI. The first method uses the mean axes of vectorcardiographic T-loops taken from the inverse Dower transform of the 12-lead ECG to indicate ischemic regions of the left ventricular wall. The second method establishes regional scores for residual ST elevation supportive of ischemia or infarction. These 2 ST-T measures qualify borderline QRS infarct criteria, resulting in composite criteria having higher sensitivities and specificities than QRS criteria alone. Comparative results were created by studying an MI positive group of 360 patients with biochemical marker evidence for infarction and an MI negative group of 515 patients negative for risk factors, biochemical markers and/or coronary disease or wall motion abnormality. Three automated ECG methods that rely predominantly on QRS markers for infarction are compared to Cardiovise (CV) prior MI algorithm version 3.0: Cardiovise 3.0 also introduces criteria for detection of acute myocardial infarction (AMI). Two automated ECG methods are compared to Cardiovise for the detection and labeling of ST elevation (STE) AMI, and for detection of non-STE AMI. PMID- 12539107 TI - Improved electrocardiographic detection of left ventricular hypertrophy. AB - We studied the echocardiographic measurements and the digital electrocardiograms (ECGs) of 3378 participants in the Cardiovascular Health Study and divided the subjects into a Learning Set of 1678 subjects and a demographically matched Test Set of 1700 subjects. We defined left ventricular hypertrophy (LVH) as values of the echocardiographic left ventricular mass index (LVMI) >116 g/m(2) in men or >104 g/m(2) in women. We performed univariate linear regression separately on the male and female subjects in the Learning Set to find all the ECG parameters that correlated significantly with LVMI. We then applied multivariate linear regression (MLR) to these parameters to identify the 4 variables for each sex that discriminated best between the subjects with and without LVH. We then applied the resultant sex-specific linear regressions to the subjects in the Test Set. We found that the diagnostic performances for LVH of the new MLR equations were superior to those of three commercial computerized ECG diagnostic algorithms in both men and women. We also found that for the new MLR and for all three commercial algorithms, the diagnostic sensitivities at matched specificities were superior in the men compared to those in the women. We conclude that the new MLR equation identifies patients with echocardiographically determined LVH more reliably than do 3 commercial computerized diagnostic algorithms and that the sensitivities of each of these modalities is greater in men than it is in women. PMID- 12539108 TI - Risk stratification applied to CAST registry data: combining 9 predictors. Cardiac Arrhythmia Suppression Trial. AB - Over 200,000 people in the United States die of sudden cardiac death (SCD) every year. Although many of these deaths occur in asymptomatic individuals, the vast majority of deaths occur in people who are under care for existing coronary heart disease. Implantable cardioverter/defibrillators (ICDs) have been shown in several randomized trials to be effective in prolonging lives of those at high risk for sudden cardiac death, but the criteria used in these trials and the ACC/AHA consensus guidelines would cover only a minority of patients. Developing methods to assign risk to individual patients without prior SCD events could promote the use of this life-saving therapy in those with especially high risk. Given sufficient physiologically relevant measurements from electrocardiogram analysis, clinical assessment, and demographic status, multivariate statistical methods for predicting survival can be used to combine many predictors of risk and calculate the risk for an individual patient. A survival analysis using Cox regression on data from the Cardiac Arrhythmia Suppression Trial (CAST) illustrates this concept. Patient age, sex, ejection fraction, smoking history, and prior myocardial infarction history, along with the frequency of premature beats and the presence of runs of ventricular tachycardia on Holter monitoring and the time from the index myocardial infarction to the baseline Holter and to recruitment into CAST were combined in a multivariate predictor derived from the Cox regression; this predictor significantly outperforms the individual predictors. A proposed test based on this predictor would identify as positive 7% of the CAST registry, with an average risk of death among the positives of 47%; 20% of those dead at 2 years would be positive. With improved component measurements, this approach has the potential for significantly improving risk stratification for the prevention of SCD. PMID- 12539109 TI - The centennial of the Einthoven electrocardiogram. AB - Recording of the electrocardiogram by string galvanometer evolved a century ago from wide-ranging advances in physiology, physics, and engineering. Although body surface recordings had been obtained prior to the work of Willem Einthoven, his ingenuity in mathematically correcting capillary electrometer tracings and incorporating recent advances in electromagnetism, recording technology, optics, and quartz chemistry into the new instrument led to a fundamental tool for the investigation of the heart. For his work, Einthoven received the Nobel Prize. PMID- 12539110 TI - Implantable loop recorders in pediatric practice. AB - Symptoms of syncope and palpitations are not uncommon in children with and without heart disease. They present a diagnostic dilemma when conventional cardiovascular testing is inconclusive. The implantable loop recorder (ILR) has been shown to play an important role in diagnosing recurrent syncope in adult patients. In pediatric practice its role still has to be defined. The aim of this review is to assess the diagnostic yield of the ILR in children and young adults. Seven patients, 4 male and 3 female, were included in the study. Mean age at implantation was 12.8, range 0.8 to 25.9. Indications for ILR were syncope (N=6), near-syncope (N=1), palpitations (N=2) and acute life threatening event (N=2). Previous testing included 12-lead ECG (N=7), echo (N=7), 24 hour Holter (N=7), 4 week ambulatory Holter (N=3), exercise test (N=4) and invasive electrophysiologic study (N=3). Over a mean follow-up period of 7.5 months (range 3 to 16 months), 4 (57%) patients continued to have symptoms. The ILR enabled the correct diagnosis to be established in all, allowing for appropriate therapy. PMID- 12539111 TI - A review of the theoretical and experimental bases of transesophageal atrial pacing. AB - Transesophageal atrial pacing has been widely used for studies in cardiac electrophysiology as well as for other cardiac studies in which acceleration of heart rate cannot be achieved or closely controlled with exercise. However, the theoretical basis of the technique and the possibility that it may produce electrical burn injury to the esophagus have not been well-described in the electrocardiographic literature. This review examines the current density produced by an esophageal electrode and compares it with that produced by an intra-atrial catheter electrode. It also presents data on the extent of esophageal burning produced by transesophageal pacing in dogs. PMID- 12539112 TI - Transesophageal electrocardiography and atrial pacing in children. AB - The technique and spectrum of clinical application of transesophageal electrocardiography and atrial pacing in children is demonstrated in this article. Special emphasis is laid on the usefulness of the method in newborns and infants for the management of supraventricular tachycardia. PMID- 12539113 TI - An esothoracic electrode for electrophysiological studies. AB - A flexible polythene esothoracic electrode was passed into the stomach under light sedation. Five ring electrodes, now positioned in the lower esophagus, were used for bipolar atrial pacing and recording. Ventricular pacing was performed using a cathodic point source on the esothoracic electrode tip; the indifferent electrode (anode) was a high impedance chest pad. Parameters of atrioventricular (AV) nodal function were obtained by atrial pacing. Programmed ventricular stimulation was performed using a standard protocol. Electrophysiological parameters were subsequently determined using an endocardial approach. There was close correlation between measurements of AV node function in 48 subjects: AV Wenckebach Cycle Length (AVWCL) r(2) =.97. The degree of agreement was estimated by the mean difference delta and standard deviation of the difference sigma (AVWCL sigma = 7 ms, delta = 16 ms). Programmed ventricular stimulation was performed in 15 subjects with known or suspected ventricular tachyarrhythmias. In subjects where ventricular tachycardia was induced by using both esothoracic and endocardial approaches, the rate, morphology, and haemodynamic consequences were similar. In several cases, ventricular tachycardia was pace-terminable using overdrive esothoracic ventricular pacing. Electrophysiological parameters determined with this electrode were similar to those obtained with endocardial stimulation. PMID- 12539114 TI - Termination of atrial reentrant tachycardias by using transesophageal atrial pacing. AB - Atrial reentrant tachycardia (ART) is not an uncommon complication in patients after surgery for congenital heart defects, particularly after extensive atrial procedures with sutures lines or baffeling maneuvers. Primary atrial flutter is a rare dysrhythmia in newborns. Two issues of ART or atrial flutter can be addressed by the esophageal approach: First, ART (often with 2:1 A-V conduction) may be difficult to detect on the surface ECG as the P waves usually do not show the typical flutter morphology or are hidden in the T wave. Recordings over the esophageal lead clearly determine the underlying rhythm. Second, termination of ART by pacing the atria can easily be performed by using an esophageal electrode. We performed 62 conversions of ART in 39 pts. (median age 12.5 years), among them 7 newborns with typical atrial flutter. The pacing algorithm started with 4 extrastimuli and pacing intervals 20 msec shorter than the atrial cycle length of tachycardia. Conversion was achieved in 50/62 (81%) cases; in 12 of 62 (19%) cases conversion was performed externally as the transesophageal approach was insufficient to restore sinus rhythm. Intravenous infusion of amiodarone prior to atrial pacing seems to be helpful in some patients for termination of ART. We recommend transesophageal atrial pacing as a first step of acute management of atrial flutter and other forms of ART in neonates, infants, and children because it is a minimal invasive procedure with high success rates. PMID- 12539115 TI - Invasive electrophysiology in children: state of the art. AB - While noninvasive techniques, including esophageal recording and pacing, clearly have advantages in the pediatric population; they also have significant limitations. Invasive electrophysiology (EP) in children now encompasses the use of many advanced engineering applications, which contribute to the efficacy of such procedures. This is particularly true with respect to the performance of radiofrequency catheter ablation. First, microcatheters (eg, Cardima) offer advantages as diagnostic catheters in small hearts, and allow pacing and recording from both atrium and ventricle in even the smallest infants using limited venous access. In addition, there is the possibility of mapping the distal coronary sinus, and the right atrioventricular groove via the right coronary artery. Second, electroanatomic mapping using the CARTO system (Biosense Webster) allows complex maps to be constructed in patients with congenital heart disease and a history of cardiac surgery. Although somewhat laborious to construct, such maps offer great detail to guide ablation, and also provide voltage information to guide identification of patches and scars. Similarly, non contact mapping systems (eg, EnSite, Endocardial Solutions) are available which record far-field potentials and employ solutions to the inverse problem to reconstruct endocardial potentials. Three-dimensional mapping based on a single beat is possible, and the success of ablation in creating conduction block in unique channels can be evaluated. Both of these three-dimensional mapping systems have the potential to improve outcomes in patients with complex disease undergoing ablation. Finally, patients are now undergoing combined procedures with interventional catheterization (eg, device closure, stenting) along with EP procedures (eg, ablation, device implant) in the same laboratory session. The state of the art in invasive pediatric EP increasingly involves the use of advanced technology adapted for use in pediatric and congenital heart disease applications. PMID- 12539116 TI - Solidifying the solid angle. AB - This paper evaluates the position of the solid angle in its application to modeling in electrocardiology. Particular attention is paid to the use of the solid angle for linking cardiac electric activity to the potentials observed on the body surface. In this application, the solid angle is a dominant factor in the expression of the sources during depolarization known as the uniform double layer. In the related equivalent double layer model, the contributions of the elementary sources are also expressed in terms of solid angles, their strength not being uniform. A recently developed theory allows the equivalent double layer to be applied to both depolarization and repolarization. PMID- 12539117 TI - Efficacy of 2 strategies to detect body position ST-segment changes during continuous 12-lead electrocardiographic monitoring. AB - Body positional change is the most frequent cause of false positive ST-segment monitor alarms. This study evaluates 2 strategies to distinguish ST events due to positional change from those due to transient myocardial ischemia: 1) A biosensor device to indicate body position with tick marks on the electrocardiogram (ECG) tracing, and 2) An initially recorded ECG "template" in four positions (supine, right, left, and upright) for subsequent comparison with ST alarm ECGs. A purposive sample of 96 patients on a cardiac telemetry unit was recruited to have both strategies implemented and evaluated. The biosensor correctly identified all 4 body positions in the 96 patients. A subgroup of 30 patients were continuously monitored and 9 (30%) had at least 1 ST event due to body position change, for a total of 25 false positive events. Of the 25 positional events detected by the biosensor, the ECG template method detected 18 (73%). The biosensor method is superior to the ECG template method for detecting positional ST events. However, a limitation of both strategies is that they require clinicians to be skilled in integrating such positional information to reach a correct diagnosis. PMID- 12539118 TI - Detailed definition and interobserver agreement for the electrocardiographic Sclarovsky-Birnbaum ischemia grading system. PMID- 12539119 TI - Development and validation of techniques for quantitative clinical assessment of myocardial infarction by electrocardiography and MRI. PMID- 12539121 TI - The first ISCE Board of "Trustees" overview panel session: ischemia monitoring, state of the art. International Society of Computerized Electrocardiology. PMID- 12539132 TI - Human vagal heart rate responses in warned reaction time tasks: a cross validation with simulated vagus-sino-atrial node interactions. AB - This study examines the hypothesis that human heart rate responses in reaction time (RT) tasks are mediated by changes in vagal nerve activity. Experimental results were compared to simulated heart rate (HR) responses. An animal model of vagus-cardiac sino-atrial node interactions reconstructed the cardiac beats. Hypothetical human, vagal excitation patterns were presented to the model. Simulated and human phase-dependent cardiac responses were highly similar. Three human heart rate responses were examined: anticipatory HR deceleration, stimulus evoked HR deceleration, and response related HR acceleration. Fast motor responses were associated with fast vagal withdrawal. Inhibition of a motor response was associated with delayed onset of vagal withdrawal. However, the nature of the motor conditions did not affect the slope of vagal withdrawal. Preceding stimulus-related vagal excitation may be partly masked in the HR response, when a fast motor response and early vagal withdrawal follow. In RT tasks, a timekeeping response to the repeatedly presented and temporally regular RT signals is assumed which is under functional control of higher brain centers. Anticipatory vagal excitation that starts some time before the RT signal and continues till the RT signal, may be related to this timekeeping response. PMID- 12539133 TI - Neurocardiological basis for intraindividual ECG variability. AB - Various combinations of 1) cardiac preload, 2) sympathetic outflow to the heart, and 3) parasympathetic outflow to the heart may yield the same heart rate, while the resulting ventricular electrophysiological properties differ. This may be an important source of intra-individual electrocardiogram (ECG) variability. Isometric stress, by invoking the ergoreflex, increases sympathetic outflow. By an increased peripheral resistance this increases cardiac afterload, activating the arterial baroreflex. This reflex, in turn, reduces the sympatho-excitation and increases parasympathetic outflow. Progressive gravitational stress (sitting standing) causes a progressively decreasing cardiac preload. Decreasing cardiac preload activates the cardiopulmonary baroreflex. This causes an increase of sympathetic outflow, while parasympathetic outlow changes in opposite direction. An example of intra-individual ECG variability in the sitting position is given for healthy men. Two stressors were applied: handgrip (ergoreflex) with the legs in horizontal position, and progressive leg lowering (gravitational load). Thirteen subjects had a heart rate match better than 1%. QRS azimuth, QRS duration, maximal T vector, T azimuth, T elevation, ST duration, QRS-T angle and QT interval differed significantly between handgrip and leg lowering (QT interval 435 +/- 21 vs. 418 +/- 15 ms, P <.01). More extreme experimental conditions and/or more sensitive substrates than healthy young men are likely to entail larger ECG differences. It may be clinically relevant to further explore this source of intra-individual ECG variability. PMID- 12539134 TI - Heart rate variability parameters correlate with functional independence measures in ischemic stroke patients. AB - This study gives quantitative information regarding the effect of brain infarction on the regulation of the cardiovascular system. Electrocardiograms of 16 patients, ranging from 54 to 85 years old, admitted for rehabilitation after an ischemic supratentorial stroke, all without a history of cardiac disease or rhythm disturbances, were recorded during physiotherapy effort. All patients were evaluated for the functional independence measure (FIM) score 48 to 72 hours following admission and repeated at one week before discharge. The heart rate variability (HRV) parameters: standard deviation (SD) of the RR series, the power spectrum of the RR intervals for the low frequency range (LF: 0.04-0.15 Hz), and the high frequency range (HF: 0.15-0.4 Hz) were calculated. The results showed that all heart rate variability parameters had statistically significant relationship with the FIM on admission and discharge scores. In conclusion, the HRV parameters correlate with the clinical measures of function: the greater the HRV parameter the higher the FIM score. The presented technique may prove useful as a prognostic tool providing a simple way for determining functional performance of stroke patients. PMID- 12539135 TI - Direct mechanical stimulation of brainstem modulates cardiac rhythm and repolarization in humans. AB - Natural mechanical stimulation of the brainstem area by the blood pressure waves propagating in the adjacent arteries plays an important role in the homeostasis of the brainstem centers of cardiovascular control. However, effects of direct mechanical stimulation of this area on the cardiac elecrophysiology have never been studied in humans. In 12 patients (age: 54 +/- 13 years, 5 females) undergoing microvascular decompression, the left (9 patients) or the right (3 patients) side of the ventro-lateral surface of the medulla oblongata was exposed during the surgery, and a mechanical stimulation (duration: 1 min, frequency: 1-2 Hz) of the roots of the cranial nerves and the surface of the brainstem was performed at 3-7 sites using a 2-mm metallic ball. Spatial changes in cardiac repolarization were examined using the 32-lead/192 site electrocardiographic body surface potential maps. Blood pressure was monitored using intra-arterial line. The intervals between the onset of the Q-wave and the offset of the T-wave (QTe) and between the onset of the Q-wave and the peak of the T-wave (QTp), the activation-recovery intervals (ARi), the peak T-wave amplitude, and the QRS and STT integrals were measured using custom software. During the stimulation between the caudal rootlets of the 10th nerve, the peak T-wave amplitude decreased 22% (range: 6-50%) and RR-intervals decreased from 923 +/- 190 to 794 +/- 111 ms compared to the recordings obtained before the stimulation (P =.025 and.063, respectively), whereas QTe, QTp, Ari, and the QRS- and the STT-integrals did not change. Decreased T-wave amplitudes and unchanged QT-intervals suggest that brainstem stimulation might evoke spatially inhomogenious repolarization changes. Stimulation of a localized region surrounding the caudal rootlets of the 10th nerve elicits pronounced effects on cardiac rhythm and repolarization. PMID- 12539136 TI - Electrocardiographic repolarization abnormalities in subarachnoid hemorrhage. AB - Electrocardiographic abnormalities, particularly in those waveforms representing ventricular repolarization, have been reported in subarachnoid hemorrhage. This study reports abnormalities on the initial electrocardiogram in 100 patients with SAH. Overall, one or more repolarization abnormalities occurred in 41% of patients. Analysis revealed prolonged QTc interval >460 ms in 16%, ST segment elevation in 9%, ST depression in 3%, T wave inversion in 7%, and U wave >or=100 microV in 15%. Electrocardiographic criteria for left ventricular hypertrophy were met in 14%, and 43% of those patients had no history of hypertension. Serum cardiac troponin I was elevated in 21%, and was significantly associated with QTc interval >460 ms (P <.001). Controlling for gender, those with QTc interval >460 ms were 5.5 times more likely to have elevated serum cardiac troponin I. It is concluded that repolarization abnormalities are present in a high proportion of patients with SAH. Some SAH patients also have left ventricular hypertrophy voltage unrelated to hypertension or coronary artery disease. Prolonged QTc interval after SAH is significantly related to myocardial injury, but unrelated to mortality, and there is no association between ST-T wave abnormalities and either myocardial injury or mortality. PMID- 12539137 TI - Personal reflections and future goals: presidential address in Park Hotel "De Branding" Doorwerth, The Netherlands 2002. PMID- 12539138 TI - Acute and sublethal effects of two insecticides on earthworms (Lumbricus terrestris L.) under laboratory conditions. AB - Earthworms (Lumbricus terrestris L.) were exposed to commercial formulations of endosulfan and aldicarb for 2, 7, and 15 days, and the LC(10), LC(25), and LC(50) were determined. Worms were then exposed to LC(10), LC(25), and LC(50) concentrations of endosulfan and LC(10) and LC(25) concentrations of aldicarb. The growth rate and total protein content were determined and related to endosulfan and aldicarb residues in soil and earthworms. Aldicarb was more toxic than endosulfan under the experimental conditions. The residues of endosulfan and aldicarb caused a significant reduction in the growth rate and total protein content of earthworms. The residues of endosulfan and aldicarb were monitored in soil and earthworms after 2, 7, and 15 days of exposure. The residues remaining in the soil after the experiments ranged between 37.75% and 68.54% of the applied concentration for endosulfan and between 10.13% and 67.71% of the applied concentration for aldicarb. Small amounts of both insecticides were detected in worms, and accumulation was more important for endosulfan. This study proposes the use of growth rate and total protein content as biomarkers for contamination by endosulfan and aldicarb. PMID- 12539139 TI - The freshwater cyanobacterium Lyngbya aerugineo-coerulea produces compounds toxic to mice and to mammalian and fish cells. AB - Despite a growing awareness of the presence of cyanobacterial toxins, knowledge about the ability of specific species to produce toxic compounds is still rather limited. It was the overall goal of the current work to investigate if probes derived from the freshwater species Lyngbya aerugineo-coerulea (Kutz.) Gomont, a cyanobacterium frequently found in southern Europe and not previously investigated for the presence of bioactive compounds, were capable of eliciting in vivo and in vitro toxicity. The cyanobacterial extract revealed signs of neuro as well as hepatotoxicity in mice, although these signs could not be explained by the well-known respective cyanobacterial neuro- and hepatotoxins saxitoxin and microcystin. Cytotoxicity was elicited by the cyanobacterial extract in all mammalian cell lines tested. As well, the rainbow trout liver cell line, RTL-W1, was found to be susceptible to the cytotoxic effects of the extract, although the cytotoxicity was dependent on temperature. In contrast, the cyanobacterial growth medium elicited cytotoxicity independent of temperature, leading to morphological changes indicative of alterations to the cytoskeleton. Overall, the results suggest that Lyngbya aerugineo-coerulea is an important cyanobacterium to be considered for its potential to cause health risks on environmental exposure of it to mammals and fish. Applying a combination of mammalian and piscine cell line bioassays is a unique approach that, combined with chemical analysis, could be used in the future to identify the structure and cellular mechanisms of the as yet-unknown toxic Lyngbya aerugineo-coerulea metabolites in particular and to screen cyanobacterial extracts for their toxicity in general. PMID- 12539140 TI - Hydrocarbon-induced changes to metabolic and detoxification enzymes of the Australian crimson-spotted rainbowfish (Melanotaenia fluviatilis). AB - The toxicity of petroleum hydrocarbons to marine aquatic organisms has been widely investigated; however, the effects on freshwater environments have largely been ignored. Selected biomarkers were measured in a freshwater species, the crimson-spotted rainbowfish (Melanotaenia fluviatilis). Fish were exposed to either a water-accommodated fraction (WAF) of crude oil or a dispersed crude oil water-accommodated fraction (DCWAF) for 3 days and were depurated for 14 days. Generally, biomarkers were altered following the short-term exposures but recovered after 14 days of depuration. Metabolic enzymes measured in gill tissue were citrate synthase and lactate dehydrogenase (LDH). As a result of WAF and DCWAF exposures, citrate synthase and LDH activities increased. Enzyme activities returned to control levels following depuration. Subsequent to the WAF exposure, hepatic ethoxyresorufin-O-deethylase (EROD) activity levels were higher than controls and they returned to control levels during depuration. For the DCWAF exposure, EROD was induced by a TPH (total petroleum hydrocarbons) concentration of 14.5 mg/L; however, after depuration the 14.5 mg/L TPH group had lower EROD activity than did controls. There were no changes in liver- to body-weight ratios or the histopathological organization of gill or liver tissues. As the majority of biomarkers returned to control levels after 14 days of depuration, rainbowfish were able to recover from short-term exposures to crude oil and dispersed crude oil. PMID- 12539141 TI - Identification and quantification of in vitro adduct formation between protein reactive xenobiotics and a lysine-containing model peptide. AB - Formation of in vitro adducts between different classes of xenobiotics and the lysine-containing peptide Lys-Tyr was monitored by high-performance liquid chromatography and electrospray ionization mass spectrometry. The molecular structures of the main resulting products could be sensitively analyzed by mass spectrometry (flow injection analysis), enabling the detection of characteristic binding formations. Aldehydes such as formaldehyde, acetaldehyde, and benzaldehyde were shown to form stable linkages to lysine amino groups via Schiff bases. Other electrophilic substances (e.g., toluene-2,4-diisocyanate, 2,4 dinitro-1-fluorobenzene, 2,4,6-trinitrobenzene sulfonic acid, dansyl chloride, and phthalic acid anhydride) also formed covalent adducts with lysine residues. The reactivity of the compounds was quantified by measuring the amount of peptide that remained unchanged after incubation for a certain period with the xenobiotic. Although reactivity levels within this group of aldehydes varied only to a small extent, as would be expected, extreme differences were seen among the structurally heterogeneous group of nonaldehyde xenobiotics. These results support the hypothesis that simple chemical reactions may lead to the adduction of nucleophilic macromolecules such as peptides or proteins. Such reactions, in particular, Schiff base formation of aldehydes, have previously been shown to be capable of specifically interfering with costimulatory signaling on T cells. Our results suggest that electrophilic xenobiotics of other classes may also inherit the capacity to exert similar effects. Forming covalent linkage to peptides may represent a possible molecular mechanism of electrophilic xenobiotics in vivo, yielding immunotoxic effects. The model utilized in this study is appropriate for monitoring the adduction of xenobiotics to basic peptides and for analyzing the resulting molecular structures. PMID- 12539142 TI - Alternative inocula as activated sludge surrogate culture for a toxicity test. AB - The use of activated sludge to assess the potential toxicity and environmental impact of chemicals and wastewaters suffers from several drawbacks related to the heterogeneity, absence of standardization, and health risk associated with this mixed-sewage population. To search for reliable testing inoculum alternatives, the performance of two commercial inocula (BI-CHEM and BIOLEN M112), a garden soil inoculum and a pure culture of Pseudomonas sp., was compared with an activated sludge inoculum (AS) in the inhibition respiration test ISO 8192-1986 (E). The respiration rates of microbial inocula were assayed for the reference compound 3,5-dichlorophenol. The acute toxicity values (IC(50)) ranged from 6.7 mg/L (Pseudomonas sp.) to 22.7 mg/L (garden soil), overlapping the expected values for activated sludge microorganisms despite the bacterial diversity. The assayed microbial inocula also showed higher reproducibility than AS and an overall similarity of catabolic profiles obtained with Biolog EcoPlates was observed between AS and some mixed inocula. These results point to the potential ability of such inocula as surrogate cultures in relevant activated sludge-based bioassays. New, well-defined, standardized, and safe tools will then be available for monitoring the ecological impact of hazardous substances and effluents, thus providing environmental protection. PMID- 12539143 TI - Tannery wastewater characterization and toxicity effects on Daphnia spp. AB - Tannery wastewater contains large quantities of organic and inorganic compounds, including toxic substances such as sulfides and chromium salts. The evaluation of wastewater quality in Chile nowadays is based on chemical specific measurements and toxicity tests. The goal of this research was to characterize tannery wastewater and to relate its physical/chemical parameters with its acute toxicity effect on Daphnia pulex. To distinguish the most important toxic compounds, physical/chemical techniques were applied to a grab sample of a final effluent based on the Phase I toxicity identification evaluation (TIE) procedure. In addition, the toxicity of a beamhouse effluent after an activated sludge reactor treatment was investigated on Daphnia magna (introduced species) and D. pulex (native species). Effluent from different tannery processes (soaking, beamhouse, tanning and final) demonstrated high values of chemical organic demand (COD; 2840 27,600 mg L(-1)), chloride (1813-16,500 mg L(-1)), sulfate (230-35,200 mg L(-1)), and total solids (8600-87,100 mg L(-1)). All effluents showed extremely toxic effects on D. pulex, with 24-h mean lethal values (LC(50)) ranging from 0.36% to 3.61%. The Phase I TIE profile showed that toxicity was significantly reduced by air stripping, filtration, and a cationic exchange resin, with toxicity reductions ranging between 46% and 76%. The aerobically treated beamhouse effluent showed significantly less toxicity for both species (43%-74%). The chemical parameters demonstrated that the remaining toxicity of the treated beamhouse effluent was associated with its ammonia (120 mg N-NH(3) L(-1)) and chloride (11,300 mg Cl(-) L(-1)) contents. PMID- 12539144 TI - Uptake and accumulation of dissolved, radiolabeled nodularin in Baltic Sea zooplankton. AB - The mass occurrence of toxic cyanobacteria is a recurrent phenomenon in the Baltic Sea. Grazers may obtain toxins either through ingestion or by direct exposure to dissolved toxins. Despite this, there is little knowledge about the accumulation of cyanobacterial toxins in planktonic organisms present during these blooms. Toxin analyses of tissue samples are complicated to carry out and, because of the small size of microscopic planktonic organisms, often difficult to execute. Therefore, we wanted to use a precise and sensitive method to study toxin uptake and accumulation in zooplankton. We used chemically tritiated nodularin, (3)H-dihydronodularin, to study the uptake of dissolved nodularin, a cyanobacterial hepatotoxin produced by Nodularia spumigena. Cultures of the calanoid copepods Acartia tonsa and Eurytemora affinis, and an oligotrich ciliate Strombidium sulcatum were exposed to (3)H-dihydronodularin in filtered seawater, using naturally occurring concentrations of dissolved nodularin (5 microg L(-1)). All three species took up measurable amounts of radiolabeled nodularin. After 48 h we detected 0.37 +/- 0.22 microg toxin g C(-1) (mean +/- sd) in A. tonsa and 0.60 +/- 0.15 microg toxin g C(-1) in E. affinis, whereas 1.55 +/- 0.50 microg toxin g C(-1) was detected in S. sulcatum after 24 h. The minimum bioconcentration factor (BCF) of (3)H-dihydronodularin was 12 for A. tonsa and 18 for E. affinis. For S. sulcatum our results indicate a maximum BCF of 22. However, because the uptake studies for this species were done in the presence of bacteria, possible particulate transfer cannot be excluded. Nevertheless, our results indicate that dissolved nodularin can be taken up by planktonic organisms. Therefore, the vectorial transport of dissolved toxins to higher trophic levels seems possible, even if some planktonic grazers would avoid feeding on toxic cyanobacteria filaments. PMID- 12539145 TI - Depuration kinetics and persistence of the cyanobacterial toxin microcystin-LR in the freshwater bivalve Unio douglasiae. AB - We carried out uptake and depuration experiments in the laboratory to investigate the effects of temperature (15 degrees C and 25 degrees C) on the depuration kinetics and persistence of a cyanobacterial toxin, microcystin-LR (MCYST-LR), in a freshwater bivalve, Unio douglasiae. Bivalves were fed toxic Microcystis cells in the 15-day uptake experiment and nontoxic diatoms in the following 15-day depuration experiment. Each bivalve's hepatopancreas was lyophilized and extracted with a butanol:methanol:water solution for analysis of MCYST-LR by high performance liquid chromatography. The toxin in the organ accumulated rapidly after the beginning of the uptake experiment and reached approximately steady state conditions on day 5 at concentrations of 130 +/- 11 microg g(-1) dry weight at 15 degrees C and 250 +/- 40 microg g(-1) at 25 degrees C. In the depuration experiments MCYST-LR was eliminated asymptotically from the tissue. The values of the depuration rate constant (k(d)), calculated with a first-order one compartment model, were found to be 0.142 +/- 0.044 day(-1) at 15 degrees C and 0.226 +/- 0.046 day(-1) at 25 degrees C. The depuration Q(10) value from 15 degrees C to 25 degrees C equaled 1.6. This study was the first to reveal the kinetics of depuration for MCYST-LR in a bivalve. The results show that MCYST-LR may be eliminated slowly in autumn and winter and persist in the tissue until spring. Thus, in terms of toxicokinetics, the risk to people of being poisoned by bivalves would increase if toxic blooms occur in autumn. PMID- 12539148 TI - Gene therapy and tissue engineering for sports medicine. AB - Sports injuries usually involve tissues that display a limited capacity for healing. The treatment of sports injuries has improved over the past 10 to 20 years through sophisticated rehabilitation programs, novel operative techniques, and advances in the field of biomechanical research. Despite this considerable progress, no optimal solution has been found for treatment of various sports related injuries, including muscle injuries, ligament and tendon ruptures, central meniscal tears, cartilage lesions, and delayed bone fracture healing. New biological approaches focus on the treatment of these injuries with growth factors to stimulate and hasten the healing process. Gene therapy using the transfer of defined genes encoding therapeutic proteins represents a promising way to efficiently deliver suitable growth factors into the injured tissue. Tissue engineering, which may eventually be combined with gene therapy, may potentially result in the creation of tissues or scaffolds for regeneration of tissue defects following trauma. In this article we will discuss why gene therapy and tissue engineering are becoming increasingly important in modern orthopaedic sports medicine practice. We then will review recent research achievements in the area of gene therapy and tissue engineering for sports-related injuries, and highlight the potential clinical applications of this technology in the treatment of patients with musculoskeletal problems following sports-related injuries. PMID- 12539149 TI - Adenovirus-mediated gene transfer to the transplanted piglet heart after intracoronary injection. AB - BACKGROUND: The advent of cardiac gene therapy in clinical practice requires a more efficient and safer myocardial gene delivery in large animals. A new approach to adenovirus-mediated intracoronary gene transfer in the piglet, using a heterotopic heart transplantation model, was designed to maximize the duration of contact between the vector and the heart in noncoronary flow conditions. METHODS: Recombinant adenoviruses harboring a nucleus-localized beta galactosidase gene under the control of a viral promoter were injected into the coronary vessels of the harvested hearts at a dose ranging from 10(10) to 2 x 10(11) pfu. The graft was maintained for 75 min in saline solution and then implanted in the abdomen of recipients. Gene transfer to allografts was evaluated 4 days after grafting by immunohistochemical and enzymatic analysis of beta galactosidase expression. RESULTS: Transgene expression was detected in all cardiac areas and up to 64, 44, 32, and 15% of positive nuclei were estimated in the left ventricle wall in four animals out of eleven. In the remaining animals, transgene expression was focally distributed, mainly in the left ventricle wall. PCR analysis revealed the presence of adenoviral sequences, albeit minimal, in exposed organs such as the liver and lung. CONCLUSIONS: This procedure demonstrated that direct intracoronary gene transfer can be achieved using an ex vivo gene transfer strategy. PMID- 12539150 TI - Analysis of site-specific transgene integration following cotransduction with recombinant adeno-associated virus and a rep encodingplasmid. AB - BACKGROUND: Recombinant adeno-associated virus (rAAV) has many advantages for gene therapeutic applications in comparison with other vector systems. One of the most promising features is the ability of wild-type (wt) AAV to integrate site specifically into human chromosome 19. However, this feature is lost in rAAV vectors due to the removal of the rep-coding sequences. METHODS: HeLa cells were transfected with a rep expression plasmid, infected by rAAV and grown with or without selection pressure. Single cell clones were generated and genomic DNA was analyzed for site-specific integration by Southern blotting analysis and fluorescence in situ hybridization (FISH). RESULTS: Transfection of HeLa cells with a rep expression plasmid followed by transduction with a rAAV vector resulted in site-specific integration of the transgene at AAVS1 on human chromosome 19 in 7 of 10 cell clones analyzed. In marked contrast, transduction of cells with rAAV alone did not result in any site-specific integration of the transgene. CONCLUSIONS: The high frequency with which the site-specific integration took place in the presence of Rep protein is comparable with the results observed with wtAAV. These results offer opportunities for the development of specifically integrating rAAV vectors. PMID- 12539151 TI - Relationship between the physical shape and the efficiency of oligomeric chitosan as a gene delivery system in vitro and in vivo. AB - BACKGROUND: Chitosans of high molecular weights have emerged as efficient nonviral gene delivery systems, but the properties and efficiency of well-defined low molecular weight chitosans (<5 kDa) have not been studied. We therefore characterized DNA complexes of such low molecular weight chitosans and related their physical shape and stability to their efficiency as gene delivery systems in vitro and in vivo. METHODS: Individual complexes between six different chitosan oligomers (6-, 8-, 10-, 12-, 14- and 24-mers) and fluorescence-labeled T4 DNA were visualized and classified into six physical shapes using video enhanced fluorescence microscopy. The effects of chitosan chain length, charge ratio (+/-) and solvent properties (pH and ionic strength) on the stability and structure of the complexes were studied. Gene expression in vitro and in vivo were studied using a luciferase reporter gene. RESULTS: Free DNA appeared as extended coils. Chitosan complexes had a variety of physical shapes depending on the experimental conditions. In general, the fraction of complexes that had nonaggregated, globular structures increased with increasing chain length of the chitosan oligomer, increasing charge ratio and reduction of pH (from 6.5 to 3.5). A further increase in charge ratio for globular complexes or a further reduction in pH (to 2.5) increased the fraction of aggregates, indicating a window where pharmaceutically desirable globules are obtained. Gene transfection efficiencies in vitro and in vivo were related to the physical shape and stability of the complexes. Only the 24-mer formed stable complexes that gave a high level of gene expression comparable to that of high molecular weight ultrapure chitosan (UPC) in vitro and in vivo. CONCLUSIONS: Chitosan oligomers form complexes with DNA in a structure-dependent manner. We conclude that the 24-mer, which has more desirable physical properties than UPC, is more attractive as a gene delivery system than the conventional high molecular weight chitosans. PMID- 12539152 TI - Uptake by mouse liver and intracellular fate of plasmid DNA after a rapid tail vein injection of a small or a large volume. AB - BACKGROUND: An efficient gene transfer can be achieved in mouse liver by a rapid tail vein injection of a large volume of plasmid DNA solution (hydrodynamics based transfection). The mechanism of gene transfer by this procedure is not known. It must be related to the uptake and intracellular fate of DNA. METHODS: We have investigated the problem by following the uptake by mouse liver and the intracellular distribution of DNA after a rapid tail vein injection of a large (2.0 ml) or a small (0.2 ml) volume of (35)S-DNA solution. Total and acid-soluble radioactivity were measured in liver homogenates at increasing times after injection, and their subcellular distributions were established by centrifugation methods and compared with the distributions of marker enzymes of the membrane compartments involved in endocytosis: alkaline phosphodiesterase (plasma membrane) and cathepsin C (lysosomes). RESULTS: (35)S-DNA uptake by the liver is similar when a small or a large volume of injection is used but its degradation is markedly slower after a 2.0 ml injection. When a small volume of injection is given, distribution of radioactivity after differential centrifugation indicates that the plasmid DNA is endocytosed and reaches lysosomes where it is hydrolysed. After a large volume injection, part of (35)S-DNA has the same fate, another part remains acid-precipitable for at least 1 h and is associated with structures sedimenting at low centrifugation speed in the nuclear fraction N. Analysis of that fraction by gradient centrifugation suggests that these structures are plasma membrane fragments that could originate from the apical domain of hepatocytes. The proportion of (35)S-DNA associated with hepatocytes is about doubled after a large volume injection. Fractionation of isolated hepatocytes by centrifugation confirms results obtained on the whole liver. Treatment of the N fraction or isolated hepatocytes with pancreatic DNAse illustrates that (35)S-DNA that remains bound to plasma membrane after a large volume injection is located on the outer face. CONCLUSIONS: The fact that after an hydrodynamic injection (35)S-DNA remains bound to the outside face of the plasma membrane for at least 1 h indicates that it is not, or very slowly, internalised during that period. The relatively small difference in the amount of DNA picked up by hepatocytes depending on the type of injection could not explain the absence of expression after a conventional injection and the strong expression after a hydrodynamic injection. If DNA enters the cells by endocytosis, even after an hydrodynamic injection, its persistence at the outside face of the plasma membrane could favour transfection by allowing hepatocytes to dispose for a relatively long time of a reservoir of intact DNA. PMID- 12539153 TI - Marked enhancement in gene expression by targeting the human insulin receptor. AB - BACKGROUND: Exogenous genes can be delivered to cells without viral vectors using an "artificial virus" comprised of nonviral plasmid DNA encapsulated in the interior of 85 nm pegylated immunoliposomes (PIL). The liposomes are targeted to cells with receptor-specific targeting ligands such as receptor-specific peptidomimetic monoclonal antibodies. METHODS: The levels of luciferase gene expression in human or rat glioma cells are measured after targeting the PIL encapsulated plasmid DNA via the human insulin receptor, the human epidermal growth factor receptor, or the rat transferrin receptor. The luciferase expression plasmids were either derived from pCEP4, which contains the Epstein Barr nuclear antigen-1/oriP replication system, or from pGL2, which lacks this system for episomal replication of plasmid DNA. RESULTS: Depending on the plasmid construct used and the receptor targeted, the peak luciferase gene expression varied more than 200-fold from 1.8 +/- 0.1 to 419 +/- 31 pg luciferase per mg cell protein. With the same plasmid, the peak level of gene expression following delivery to the cell via the human insulin receptor was 100-200-fold higher than gene expression following delivery via either the epidermal growth factor receptor or the transferrin receptor. There was no gene expression if the targeting ligand on the PIL was replaced with a nonspecific isotype control antibody. CONCLUSIONS: The extent to which an exogenous gene is expressed within a cell via a nonviral, receptor-mediated gene transfer technology is determined by the receptor specificity of the targeting ligand. The highest levels of gene expression are obtained after targeting the insulin receptor, and this may derive from the nuclear targeting properties of this receptor system. PMID- 12539154 TI - Efficacy of interleukin-10 gene electrotransfer into skeletal muscle in mice with collagen-induced arthritis. AB - BACKGROUND: Gene therapy is very promising in the treatment of rheumatoid arthritis (RA). Electrotransfer is a recent method reported to enhance in vivo intramuscular DNA transfection. Interleukin-10 (IL-10) has antiinflammatory effects in RA and in collagen-induced arthritis (CIA), a murine model of RA. In order to improve our strategy of gene therapy, we used electrotransfer to enhance penetration into skeletal muscle with CIA of plasmids encoding IL-10. METHODS: CIA was induced in DBA/1 mice by immunization with bovine type II collagen. Injection into the tibial cranial muscle of low-dose (200 ng) pCOR plasmid encoding murine IL-10 (pCOR-CMV-mIL-10) was immediately followed by application of square-wave electric pulses (8 pulses of 200V/cm, 20 ms duration at 2 Hz). Control groups received empty plasmid or saline before electrotransfer. RESULTS: When electrotransfer was performed twice on days 10 and 25 postimmunization, CIA was significantly delayed (P < 0.05) and attenuated (P < 0.001) in groups treated by electrotransfer or pCOR-CMV-mIL-10 plasmid vs. control groups. When electrotransfer of pCOR-CMV-mIL-10 plasmid was performed on days 25 and 40 postimmunization, at disease onset, the clinical severity of CIA was reduced (P < 0.05). All groups which had been electrotransferred early or late by pCOR-CMV-mIL 10 plasmid showed suppression of histological signs of arthritis. CONCLUSIONS: Taken together, these data indicate that administration of an antiinflammatory plasmid-born gene by electrotransfer of naked DNA is effective in vivo in an arthritis model. PMID- 12539156 TI - Architectonics of crayfish ganglia. AB - The central nervous system of crayfish consists of a chain of segmental ganglia that are linked by cables of intersegmental axons. Each ganglion contains a highly-ordered core of longitudinal tracts, vertical tracts, commissures, and synaptic neuropils. We review from a technical perspective the history of the description of these ganglia, and recognize four episodes of progress. Each major innovation in anatomical methods has led to new insight into the structure and function of this nervous system, and new awareness of the structural patterns that are common to the CNS of all arthropods. Ganglia in different segments of the body differ in size, and appear to differ in anatomy. From a comparison of the structures of the cores of abdominal, thoracic, and subesophageal ganglia, we argue that this apparent difference is illusory. Rather, each of these ganglia is organized on the same plan, a plan also found in insect segmental ganglia. The apparent differences follow from longitudinal compression during development and from allometric growth of particular neuropils associated with innervation of the walking legs. Different authors have described the internal organization of ganglia in different segments, so we provide a cross-reference to the nomenclatures they have introduced. We compare the locations of cell bodies of motor neurons and accessory neurons that innervate different peripheral structures, and demonstrate double-labeling of certain GABAergic peripheral inhibitory neurons. Finally, we describe the construction of digital movies of serial sections of these ganglia, and discuss their utility. PMID- 12539157 TI - Development, growth, and plasticity in the crayfish olfactory system. AB - Decapod crustaceans have a well-defined olfactory system characterised by a set of chemosensitive sensilla grouped together in an array (the olfactory organ) on their antennules. Olfactory receptor neurons in the olfactory organ project exclusively to, and terminate in, cone-shaped olfactory glomeruli in a discrete neuropil in the brain, the olfactory lobe. The olfactory organ appears to be the only afferent input to the olfactory lobe, making the system convenient for the study of its development and growth. The progression of development of the olfactory system is a continuum and can be traced from the first appearance of peripheral receptor cells and central stem cells through to the construction of the tracts and neuropils that constitute the adult system. Cell proliferation leading to the production of peripheral and central olfactory neurons can be observed with mitotic markers in both embryonic stages and in postembryonic growth. Cell proliferation in the olfactory system in crayfish persists throughout the lives of the animals and can be modulated by manipulating the living conditions imposed on growing animals. Large serotonergic neurons that are associated with the olfactory system may play a role in the regulation of cell proliferation. PMID- 12539158 TI - Active dendritic properties constrain input-output relationships in neurons of the central olfactory pathway in the crayfish forebrain. AB - Parasol cells are multimodal sensory interneurons of the hemi-ellipsoid body in the decapod forebrain. In reptant crustaceans, the hemi-ellipsoid body resides in the base of the eyecup, as an appendage to the terminal medulla. Parasol cells exhibit periodic depolarizations at a frequency of 0.5-1.0 Hz. I have investigated the role of these periodic depolarizations and their superimposed impulse bursts in affecting the input/output properties of these neurons. Parasol cells receive input from photic, olfactory, and mechanosensory pathways. Strong stimulation over any one of these pathways can lead to the generation of one or more impulse bursts in a subset of parasol cells, timed to occur at the peak of successive periodic depolarizations. A role for the periodic depolarizations in the function of the parasol cells has yet to be established. I suggest the possibility that they may act as a nonlinear amplifier that boosts spatially summated excitatory synaptic potentials from strong or appropriate stimuli above threshold for burst generation. Another possibility includes modification of voltage-sensitive ion channels in the dendritic membrane, permitting a more effective spread of excitatory synaptic currents to impulse or burst initiating zones. Impulse bursts may be a highly effective mode of output for these neurons, especially so as they occur synchronously in a subset of cells in response to strong sensory input. Furthermore, backfiring of bursts into the dendritic tree has a brief (2-3 second) but effective suppressive action upon weak sensory input, which can thereby be masked by stronger, burst-generating input. This masking phenomenon is seen in other arthropod sensory interneurons, where its physiological basis appears to be a transient accumulation of intracellular Ca(++) ions that open calcium-sensitive potassium channels. PMID- 12539159 TI - The circadian system of crayfish: a developmental approach. AB - Adult crayfish exhibit a variety of overt circadian rhythms. However, the physiological mechanisms underlying the overt rhythms are controversial. Research has centered on two overt rhythms: the motor activity and the retinal sensitivity rhythms of the genus Procambarus. The present work reviews various studies undertaken to localize pacemakers and mechanisms of entrainment responsible for these two rhythms in adult organisms of this crustacean decapod. It also describes an ontogenetic approach to the problem by means of behavioral, electrophysiological, and neurochemical experiments. The results of this approach confirm previous models proposed for adult crayfish, based on a number of circadian pacemakers distributed in the central nervous system. However, the coupling of rhythmicity between these independent oscillators might be complex and dependent on the interaction between serotonin (5-HT), light, and the crustacean hyperglycemic hormone (CHH). The latter compound has, up until now, not been considered as an agent in the genesis and synchronization of the retinal sensitivity rhythm. PMID- 12539160 TI - Neuropeptides in the crayfish stomatogastric nervous system. AB - Neuropeptides are peptides with profound effects on the nervous system. The function of neuropeptides can be studied in detail in the stomatogastric nervous system (STNS). Neuropeptides are ubiquitously distributed in the STNS and it contains well-studied neural circuits that are strongly modulated by neuropeptides. The STNS controls the movements of the foregut in crustaceans and has been studied intensively in a variety of decapod crustaceans including crayfish. This article reviews our knowledge of neuropeptides in the crayfish STNS. Within crayfish, peptides reach the circuits of the STNS as neurohormones released by neurohaemal organs or by putative neurohemal zones located within the STNS. As transmitters, neuropeptides are present in identified motoneurons, interneurons, and sensory neurons (mainly shown by immunocytochemistry), indicating a multiple role of peptides in the plasticity of neural networks. Neuropeptides are not only present in varicosities within the neuropil of ganglia, but also in varicosities on muscles and within small neuropil patches along nerves. This suggests that the muscles of the stomach are under a more direct modulatory control than previously thought, and that information processing can also occur within nerves. In addition to anatomical studies, biochemical and electrophysiological methods were used. For example, MALDI-TOF MS (matrix-assisted laser desorption ionization time of flight mass spectrometry) revealed the presence of four different peptides of the orcokinin family within a single neuron, and electrophysiological experiments demonstrated that the networks of the STNS are not only under excitatory but also inhibitory peptidergic influence. Comparing the similarities and differences between the STNS of crayfish and that of other decapod crustaceans has already contributed to our knowledge about peptides and will further help to unravel peptide function in the plasticity of neural circuits. For example, the identified neurons in the STNS can be used to study co-transmission because neuropeptides are co-localized with classical transmitters, biogenic amines, or other peptides in these neurons. PMID- 12539161 TI - Physiological functions of FMRFamide-like peptides (FLPs) in crustaceans. AB - Neuropeptides play important roles in chemical signalling in the central and peripheral nervous systems. One of the largest families of neuropeptides is that of the FMRFamide-like peptides (FLPs). This paper reviews what is known about the physiological functions of FLPs in crustaceans, focussing on the cardiovascular, digestive and neuromuscular systems. PMID- 12539162 TI - Dopamine in crayfish and other crustaceans: distribution in the central nervous system and physiological functions. AB - Dopamine is widely distributed in the crustacean nervous system and has a diverse array of physiological effects. Immunocytochemical studies of several species have shown that dopamine- and/or tyrosine hydroxylase-containing cells occur in all ganglia of the central nervous system and that processes from some of these cells link ganglia of the ventral nerve cord. This study describes the distribution of tyrosine hydroxylase-containing cells in the central nervous system of a crayfish (Orconectes rusticus) and compares this information to available data from other species. The distribution of tyrosine hydroxylase (an enzyme in the synthetic pathway between tyrosine and dopamine) in O. rusticus is similar to that reported for marine species. However, differences were observed in the number of neurons in some ganglia and in the axonal projections of the L cell, which were more extensive in O. rusticus than in other species studied thus far. We also review the physiological effects of dopamine in crayfish and other crustaceans, focusing on the amine's actions in the endocrine, cardiovascular, and nervous systems, and on behavior when injected into freely-moving animals. PMID- 12539163 TI - The effects of serotonin and ecdysone on primary sensory neurons in crayfish. AB - The overall behaviors and motivational states observed during social interactions and throughout the molting cycle of crayfish have been linked to the effects of humoral neuromodulators. Both serotonin (5-HT) and a molt-related hormone, 20 hydroxyecdysone (20-HE), are known to be present in the hemolymph of crustaceans. To determine if they alter the activity of a primary sensory neuron that monitors proprioceptive information, we examined their effects on the activity of the slow adapting muscle receptor organ (MRO) of the crayfish abdomen, a model sensory system that has been extensively studied. 5-HT within the range of 100 nM to 1 microM, increases the firing frequency of the neuron during sustained stimulation. In experiments in which 20-HE was added alone, an increase in the firing frequency also occurred, although to a lesser degree than that for 5-HT at the same concentrations. When the MRO is first exposed to 20-HE, followed sequentially by 5-HT, the activity increases to about the same degree as in the reverse order of exposure. This outcome indicates that mixtures of these endogenous neuromodulators, at various levels, are more important in alternating behavior than the absolute level of any one of them introduced alone. PMID- 12539164 TI - Neural basis of a simple behavior: abdominal positioning in crayfish. AB - Crustaceans have been used extensively as models for studying the nervous system. Members of the Order Decapoda, particularly the larger species such as lobsters and crayfish, have large segmented abdomens that are positioned by tonic flexor and extensor muscles. Importantly, the innervation of these tonic muscles is known in some detail. Each abdominal segment in crayfish is innervated bilaterally by three sets of nerves. The anterior pair of nerves in each ganglion controls the swimmeret appendages and sensory supply. The middle pair of nerves innervates the tonic extensor muscles and the regional sensory supply. The superficial branch of the most posterior pair of nerves in each ganglion is exclusively motor and supplies the tonic flexor muscles of that segment. The extension and flexion motor nerves contain six motor neurons, each of which is different in axonal diameter and thus produces impulses of different amplitude. Motor programs controlling each muscle can be characterized by the identifiable motor neurons that are activated. Early work in this field discovered that specific central interneurons control the abdominal positioning motor neurons. These interneurons were first referred to as "command neurons" and later as "command elements." Stimulation of an appropriate command element causes a complex, widespread output involving dozens of motor neurons. The output can be patterned even though the stimulus to the command element is of constant interval. The command elements are identifiable cells. When a stimulus is repeated in a command element, from either the same individual or from different individuals, the output is substantially the same. This outcome depends upon several factors. First, the command elements are not only identifiable, but they make many synapses with other neurons, and the synapses are substantially invariant. There are separate flexion-producing and extension-producing command elements. Abdominal flexion-producing command elements excite other flexion elements and inhibit extensor command elements. The extension producing elements do the opposite. These interactions insure that interneurons of a particular class (flexion- or extension-producing) synaptically recruit perhaps twenty others of similar output, and that command elements promoting the opposing movements are inhibited. This strong reciprocity and the recruitment of similar command elements give a powerful motor program that appears to mimic behavior. PMID- 12539166 TI - The neural basis of dominance hierarchy formation in crayfish. AB - Fifty years of study of the nervous system and behavior of crayfish have revealed neural circuits for movements that are similar to those seen during formation of a dominance hierarchy. Given this background, it is of interest to ask what is understood about the neural substrates of dominance hierarchy formation. Here we will consider the social behavior that crayfish display in the wild and in the laboratory, and its relationship to movements released by activation of specific neural circuits. We will consider how these movements might be knit together to produce the behavior patterns that are characteristic of dominant and subordinate animals. PMID- 12539167 TI - Endothelium and valvular diseases of the heart. AB - It has become increasingly evident that the endothelium plays a critical role in the pathogenesis of valvular heart disease. The endothelium helps regulate vascular tone, inflammation, thrombosis, and vascular remodeling. Dysfunction of the endothelial cells has been linked to many vascular disorders including atherosclerosis. Common valvular diseases such as senile degenerative valve disease, myxomatous (or floppy) valves, rheumatic valves, and infective endocarditis valves show changes in the synthetic, morphologic, and metabolic functions of the valvular endothelial cells. These diseases are active processes related to endothelial cell dysfunction. Endothelial cell dysfunction is caused by mechanical forces, bacterial infection, autoantibodies, and circulating modulators of endothelial cell function. This study reviews the role of endothelial cell dysfunction in the more common valvular diseases. Continued research on endothelial cell dysfunction is crucial to our understanding of valvular heart diseases and may elucidate novel treatment and prevention strategies. PMID- 12539165 TI - Amine neurochemistry and aggression in crayfish. AB - A primary goal of our research is to explore proximate mechanisms important in recruiting adaptive social behaviors. For instance, if one of three different behaviors may be expressed in a particular set of circumstances, how do neurochemical mechanisms bias behavior towards the expression of one act in lieu of the other possibilities? In this article, we review recent results suggesting that serotonin may play such a role in the control of aggression in crayfish. First, we summarize techniques that have been optimized for sensitive characterization of neurochemical profiles in crayfish. Then, borrowing concepts from behavioral ecology, we review a framework for quantitative investigation, which regards behavior as a set of individual decisions, each with a particular probability for occurrence, a motivational context, and controlled by its own distinct neurochemical mechanisms. PMID- 12539168 TI - Angiogenesis of the heart. AB - Despite continued advances in the prevention and treatment of coronary artery disease, there are still a large number of patients who are not candidates for the conventional revascularization techniques of balloon angioplasty and stenting, or coronary artery bypass grafting (CABG). Therapeutic angiogenesis, in the form of the administration of growth factor protein or gene therapy, has emerged as a promising new method of treatment for patients with coronary artery disease. The goal of this strategy is to promote the development of supplemental blood conduits that will act as endogenous bypass vessels. New vessel formation occurs through the processes of angiogenesis, vasculogenesis, and arteriogenesis, under the control of growth factors such as those that belong to the vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF) and angiopoeitin (Ang) families of molecules. Preclinical studies have suggested that such an approach is both feasible and effective; however many questions remain to be answered. This review will address the elements of pharmacologic revascularization, focusing on gene and protein-based therapy. The important growth factors, the vector (for gene therapy), routes of delivery, the desired therapeutic effect, and quantifiable clinical end points for trials of angiogenesis will all be addressed. PMID- 12539170 TI - Endothelial cell tumors. AB - Historically, there has been much debate on the nature of infantile hemangiomas as either congenital malformations or benign neoplasms. Some vascular lesions that are present at birth and evidence no proliferative growth are considered to be congenital malformations; other post-natal vascular tumors pursue aggressive and possibly lethal clinical courses. The literature of the last two decades has been reviewed with a hope of clarifying the pathogenesis and underlying molecular lesions of this diverse set of lesions. Genetic investigations of two diseases associated with vascular tumors and abnormalities, von Hippel-Lindau disease, and Hereditary Hemorrhagic Telangiectasia have greatly added to our knowledge of vascular proliferation and provided a tantalizing clue to the pathogenesis of hemangioblastomas. Mutations have also been described in infantile hemangiomas. All of the entities considered, vascular neoplasms as well as malformations, have been examined for the expression of vascular growth factors, their receptors, and factors that appear to promote cell proliferation. Similarly, factors that either block or promote apoptosis have also been examined in various vascular lesions. These studies have in large confirmed our expectations about proliferating tumors that show upregulation of growth promoting factors and inhibition of those that promote apoptosis. In conclusion, although much has been learned about vascular physiology and the control of endothelial proliferation, and while understanding about the molecular pathogenesis of the two inherited diseases mentioned above is detailed but not yet complete, understanding of the pathogenesis of benign and malignant endothelial tumors remains vague. PMID- 12539169 TI - Von Hippel-Lindau disease. AB - Von Hippel-Lindau (VHL) disease is an uncommon, autosomal dominant hereditary multitumor syndrome caused by germline alterations of the VHL gene, which has been cloned recently and identified as a tumor suppressor gene. The major lesions in VHL disease include hemangioblastomas in the central nervous system and retina, clear cell renal cell carcinomas, pheochromocytomas, pancreatic tumors, epididymal cystadenomas, endolymphatic sac tumors, carcinoid tumors, and multiple cysts of the kidney, pancreas, and epididymis. Compared with sporadic ones, the tumors in VHL disease develop at an earlier age and often multifocally. Histologic features of VHL tumors are characterized by their high degree of vascularization and the presence of a clear cell component. Hypervascularization is induced by overexpression of vascular endothelial growth factor (VEGF), and because the principal function of VHL protein is the negative regulation of hypoxia-inducible mRNAs including VEGF mRNA, inactivation of VHL gene plays critical roles in angiogenesis of the VHL tumors. In addition, since VHL protein is also required for the down-regulation of transcription activity of certain genes for the cell growth and cell cycle, inactivation of VHL gene may contribute to tumorigenesis of the VHL tumors. A significant difference in the frequency of types of VHL gene mutation has been noted among the affected families, known as the genotype-phenotype correlations. PMID- 12539171 TI - Lymphangiogenesis in tumors: what do we know? AB - Lymphangiogenesis, the growth of new lymphatic vessels, has long been regarded as a putative efficient pathway to neoplastic metastization. However, until recently consistent data regarding reliable lymphatic endothelial cell markers were lacking. Moreover, the presence of new formed lymphatic vessels was considered a largely disputable concept. Now, this scenario has changed significantly, owing to consistent reports describing novel lymphatic endothelial cell (LEC) markers, the demonstration of new formed lymphatic vessels within the bulk of the tumor in animal models and human neoplasms, and the characterization of the VEGF-C/VEGFR-3 pathway. We herein review the major breakthroughs in the field of lymphangiogenesis, with special emphasis on novel and reliable LEC markers, such as prox-1, LYVE-1, and podoplanin, as well as on the pathological assessment of lymphangiogenesis as a putative prognostic factor for human neoplasms. PMID- 12539172 TI - Angiogenesis in endocrine glands: special reference to the expression of vascular endothelial growth factor. AB - Recent studies have shown that several angiogenic growth factors are produced and secreted by normal endocrine cells and are increased in pathological states of endocrine glands, including inflammation, hyperplasia, and neoplasia. Expression of corresponding receptors on epithelial cells and/or endothelial cells enables these angiogenic factors to influence growth and function of the endocrine tissues by auto- or paracrine mechanisms. Some of the angiogenic factors are also considered to be involved in angiogenesis, which is a critical process in tumor formation and progression. Vascular endothelial growth factor (VEGF) is regarded as one of most important angiogenic factors with specific effects on endothelial cell growth and vascular permeability, and is isolated from a variety of normal and neoplastic endocrine cells. In this article, recent studies on angiogenic factors, especially on expression of VEGF, are reviewed in the field of endocrine systems. PMID- 12539173 TI - Tumor angiogenesis of non-small cell lung cancer. AB - Lung cancer is one of the commonest causes of cancer death in developed countries. Recent evidence suggests that angoigenesis is related to poor prognosis in many solid tumors including non-small cell lung cancer (NSCLC). Angiogenesis is regulated by a complex interaction among growth factors and cytokines and influenced by proteolytic enzymes such as plasminogen activators and matrix metalloproteases, expression of adhesion molecules, and distribution of extracellular matrices. Fibroblasts, macrophages, mast cells, and endothelial cells themselves also affect angiogenesis. This review concentrates on angiogenic growth factors including vascular endothelial growth factor, angiopoietins, platelet derived endothelial growth factor, and basic fibroblast growth factor, proteases, adhesion molecules including vascular endothelial cadherin and integrins, osteopontin, and mast cell products in tumor angiogenesis of NSCLC. PMID- 12539174 TI - Biology of angiogenesis in tumors of the gastrointestinal tract. AB - The realization that the growth and spread of tumors are dependent on angiogenesis has created new avenues of research designed to help us to better understand cancer biology and to facilitate the development of new therapeutic strategies. However, the process of angiogenesis consists of multiple, sequential, and interdependent steps with a myriad of positive and negative regulators of angiogenesis being involved. The survival of tumors and thus their metastases are dependent upon the balance of endogenous angiogenic and anti angiogenic factors such that the outcome favors increased angiogenesis. Several growth factors have been identified that regulate angiogenesis in cancers of the gastrointestinal tract. These include pro-angiogenic factors like vascular endothelial growth factor (VEGF) and anti-angiogenic factors, i.e., thrombospondin. The following review provides a brief overview about the most important factors that are involved in the angiogenic process in tumors derived from colon, stomach, and pancreas. A thorough understanding of the role these factors play in the angiogenic process may lead to the development of novel therapeutic antineoplastic strategies. PMID- 12539175 TI - Angiogenesis in cutaneous melanoma: pathogenesis and clinical implications. AB - Neovacularization is an essential step in the multistage progression of malignant melanoma. The onset of new blood vessel formation is ushered in by the release of VEGF and numerous other angiogenic molecules by the tumor cells. Human melanoma is unique among neoplasms that both avascular (early horizontal growth phase characterized by very slow progression and 99%, 10-year survival) and vascular (late radial and vertical growth phase associated with rapid growth, metastasis and death in many cases), phases are discernible by the naked eye. Although cell biologists have made great strides in unraveling the mechanisms involved in the laying down of tumor vasculature and the factors that inhibit it, clinicians treating melanoma have been rather slow to realize and utilize the full potential of suppressing the tumor blood flow to the best advantage of the patient. We suggest a consorted endeavor by all the melanoma experts across the globe to establish an "angiogenesis database" wherein they pool the blood flow and vascularity information along with Breslow's thickness, Clark's level of invasion, lymphatic and vascular invasion, regression, and outcome of their patients. PMID- 12539176 TI - Angiogenesis in brain tumors. AB - Angiogenesis is a complex process regulated by multiple stimulatory and inhibitory factors that are able to modulate the migration and/or proliferation of microvascular cells with the objective of formation of neovasculature from preexisting vessels. It involves well-coordinated steps including production and release of angiogenic factors, proteolytic degradation of extracellular matrix components to allow formation of capillary sprout, proliferation and directional migration of microvascular cells, and the final composition of new vessels [Senger (1996) Am. J. Pathol. 149:1-7]. Angiogenesis is present in a number of hypoxic and/or ischemic conditions in the central nervous system, in particular in infarctions and infectious processes. Angiogenesis also plays an important role in malignant primary tumors. Glioblastomas, the most malignant gliomas in adults, are among the most angiogenic of all human tumors. This review will examine recent data of the role of angiogenic growth factors in the neoplastic and reactive conditions in the brain. PMID- 12539177 TI - Vascular networks and endothelial cells in the rat experimental pituitary glands and in the human pituitary adenomas. AB - There has been considerable interest in the relationship between hormone secreting endocrine cells (HSEC) and their microvessels (MVN) in human pituitary gland. However, microcirculatory networks have rarely been studied in three dimensions (3D). Therefore, this study was designed to visualize and to reveal the relationship between hormone secreting endocrine cells and their microvessel environment including vascular endothelial cells in 3D using rat pituitary glands under various experimental conditions by confocal laser scanning microscopy (CLSM). By CLSM, the 3D distributions of MVN were visualized and revealed a relationship between HSEC and MVN in experimental pituitary glands and human pituitary adenomas. Therefore, 3D reconstructed imaging by CLSM is a useful technique with which to investigate the microvessel environment of hormone secreting cells and has the potential to reveal dynamic hormone-secreting pathways. PMID- 12539178 TI - Angiogenesis in pituitary adenomas. AB - Angiogenesis, which is the formation of new blood vessels from preexisting capillaries, plays an important role in tumor growth and metastasis. In this review, the focus is on angiogenesis in pituitary adenomas. Angiogenesis of pituitary adenomas has been assessed by studying tumor microvessel density using specific immunohistochemical markers to clarify the relationship between angiogenesis and tumor behavior. Unlike other organs, pituitary adenomas have significantly lower vascular densities as compared to nontumorous adenohypophysis, suggesting that the lack of significant angiogenesis may play a role in the slow pace of pituitary tumor growth and rarity of metastases. In addition, the relationship between microvessel density and various factors in pituitary adenomas is reviewed, including tumor types, age and sex, invasiveness, malignancy, several proliferative markers (MIB-1 or Ki-67). However, further studies will be needed, since many studies have reached opposite conclusions. Angiogenesis is a complex multistep process and several factors are found to be involved in each step of neovascularization, such as vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and various other cytokines. VEGF and bFGF (or FGF-2), which are the most potent angiogenesis inducers among them, have been studied by immunohistochemistry, in situ hybridization, or in vitro studies in normal or adenomatous pituitaries and these results are also discussed in this review. PMID- 12539179 TI - Angiogenesis in normal and neoplastic pituitary tissues. AB - Angiogenesis, or the formation of new blood vessels, is a dynamic process needed for embryogenesis, post-natal growth, morphogenesis, tumorigenesis, and for other biological processes. Angiogenesis is very important for tumor development and progression. This review examines the activators and inhibitors of angiogenesis with emphasis on the pituitary gland and pituitary neoplasms. Some of the proteins regulating angiogenesis in pituitary tumors such as vascular endothelial growth factor (VEGF) and VEGF receptors, fibroblasts growth factors (FGF), transforming growth factor beta (TGFB), interleukins, interferons, and matrix metalloproteinases (MMPs) and inhibitors of MMPs have been examined in animal and human pituitary tumor models. However, many other significant regulators of angiogenesis including angiopoietins, angiostatin, and thrombospondins have not been studied extensively in pituitary tumors to date. Newer concepts and developments in angiogenesis such as vasculogenic mimicry and gene therapy approaches to angiogenesis in cancer treatment are also discussed. PMID- 12539180 TI - Matrix-assisted laser desorption/ionization study of cationization of PEO-PPP rod coil diblock polymers. AB - Five PEO-PPP rod-coil diblock polymers were synthesized and characterized using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). By using a series of alkali chlorides as cation salts, relative cation affinities were studied. Different cation affinities and shifts in molecular weight distributions were found as a result of changing the cation type and size. Also, a study of lithium cationization resulted in determination of a recommended quantity of lithium to be used. PMID- 12539181 TI - Identification of a potent antibacterial factor isolated from bronchoalveolar lavage fluid: guanidine, N-[3-[(aminoiminomethyl)amino]propyl]-N-dodecyl-, a potential source of error in the analysis of antibacterial agents. AB - The widespread use of antibiotics in modern society has encouraged the search for new antibacterial compounds. In this laboratory investigations are being made to identify and characterise novel antibacterial peptides. With this in mind, the antibacterial properties of human bronchoalveolar lavage (BAL) fluid from sarcoidosis patients is being investigated. In this communication we report on the identification and characterisation of a highly active non-peptide antibacterial compound isolated from BAL fluid. The structure of this active compound was elucidated by high-resolution accurate mass and tandem mass spectrometry to be guanidine, N-[3-[(aminoiminomethyl)amino]propyl]-N-dodecyl-. This compound does not appear to be endogenous, and its presence in BAL fluid extracts presents a potential source of error in analysis of antibacterial agents. The biological effects of guanidine, N-[3 [(aminoiminomethyl)amino]propyl]-N-dodecyl- have not previously been described in the literature. PMID- 12539182 TI - Determination of carbocysteine in human plasma by liquid chromatography/tandem mass spectrometry employing precolumn derivatization. AB - A sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed to determine carbocysteine in human plasma using 2-pyridylacetic acid as the internal standard (IS). The method employed derivatization with 10 M hydrochloric acid/methanol, which significantly improved the ionization efficiency of carbocysteine. After methanol-induced protein precipitation of plasma samples, carbocysteine and the IS were derivatized and subjected to LC/MS/MS analysis using atmospheric pressure chemical ionization. The method has a lower limit of quantitation of 20 ng/mL for a 0.2-mL plasma aliquot. The intra- and inter-day precision (RSD), calculated from quality control (QC) samples, was less than 7%. The accuracy, determined using QC samples, was within +/- 1%. The method offered increased sensitivity, selectivity and speed of analysis over existing methods. The method was utilized to support clinical pharmacokinetic studies of carbocysteine in volunteers following oral administration. PMID- 12539183 TI - Quantitative characterization of differential ion suppression on liquid chromatography/atmospheric pressure ionization mass spectrometric bioanalytical methods. AB - Ion suppression is a known phenomenon in atmospheric pressure ionization mass spectrometry. These suppression effects have been shown to adversely affect the accuracy and precision of quantitative bioanalytical methods. This paper presents a simple procedure for determining the impact of differential ion suppression on bioanalytical methods that utilize atmospheric pressure ionization mass spectrometric (APIMS) detection. This procedure was applied to the assessment of two potential internal standards, and to determine selectivity issues for another analyte which was to be measured in multiple species. PMID- 12539184 TI - Application of co-eluting structural analog internal standards for expanded linear dynamic range in liquid chromatography/electrospray mass spectrometry. AB - Non-linear standard calibration curves occur frequently in liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS), necessitating the use of non-linear regression curve fitting or a reduction in assay dynamic range. Non-linearity can be minimized by the use of stable isotope internal standards, if the cause of curve bending is ionization saturation. Unfortunately, stable isotope internal standards can be difficult or expensive to synthesize. Structural analog internal standards, if sufficiently close to analytes both in chemical structures and HPLC retention times, may mimic the effect of stable isotope internal standards. Tentative experimental evidence supporting this concept is presented in this paper. PMID- 12539185 TI - Collision-induced dissociation of the phenylsilane molecular ion. AB - The high-energy collision-induced dissociation of the phenylsilane molecular ion generated by electron ionization has been investigated using tandem mass spectrometry (MS/MS). It was observed that the dissociation of the molecular ion (M(+*)) occurs mainly via [M-H](+), [M-2H](+*), and [M-3H](+), followed by two consecutive losses of C(2)H(2). The structures of the precursors for the [M CH(3)](+), [M-SiH](+), and [M-SiH(2)](+*) ions are proposed. The data suggest that the molecular ion undergoes rearrangements to several isomers prior to dissociation, including the ion containing a five-membered carbon ring. Reaction mechanisms are proposed for the dissociations via the isomeric molecular ions. PMID- 12539186 TI - Ionization energies of hypervalent Li2F, Li2Cl and Na2Cl molecules obtained by surface ionization electron impact neutralization mass spectrometry. AB - Ionization energies of hypervalent Li(2)F, Li(2)Cl and Na(2)Cl molecules detected by surface ionization electron impact neutralization mass spectrometry are reported. The ionization energies were 3.78 +/- 0.2 eV for Li(2)F, 4.93 +/- 0.2 eV for Li(2)Cl, and 4.21 +/- 0.2 eV for Na(2)Cl. The ionization energies (IE) agree with theoretical ionization energies calculated by ab initio methods, supporting the theoretical prediction that Li(2)F has a hyperlithiated configuration in which the odd electron delocalizes over the two lithiums and with photoionization measurement. The first ionization energy of Na(2)Cl was experimentally confirmed earlier and for Li(2)Cl as well.8 We have developed and used this new approach for the problem--in the present work ions were first formed by surface ionization, followed by electron attachment (neutralization). PMID- 12539187 TI - Identification of the 'wrong' active pharmaceutical ingredient in a counterfeit Halfan drug product using accurate mass electrospray ionisation mass spectrometry, accurate mass tandem mass spectrometry and liquid chromatography/mass spectrometry. AB - Methodology is presented for identifying an unknown active (pharmaceutical) ingredient (AI) in a counterfeit drug product. A range of mass spectrometric techniques, i.e., accurate mass mass spectrometry, tandem mass spectrometry (MS/MS) and liquid chromatography/mass spectrometry (LC/MS), has been employed to determine the AI in a counterfeit Halfan suspension, an antimalarial drug. In particular, use of LockSpray accurate mass MS/MS allowed identification of parts of the molecule from fragments, hence limiting the number of possible elemental compositions for the nominal mass of 278 found for the AI in the counterfeit product. The analysis of the isotope pattern observed for the protonated molecule further reduced the number of possible elemental compositions. A literature search for readily commercially available compounds of molecular formula C(12)H(14)N(4)O(2)S suggested that the AI was either sulfamethazine or sulfisomidine. An LC/MS separation of those two compounds and reference MS/MS spectra obtained for sulfamethazine and sulfisomidine led to the conclusion that the AI in the counterfeit Halfan suspension is sulfamethazine, which is an antibacterial agent. PMID- 12539188 TI - Hydrogen exchange studies on Alzheimer's amyloid-beta peptides by mass spectrometry using matrix-assisted laser desorption/ionization and electrospray ionization. AB - The conformation and aggregation behavior of synthetic Alzheimer's amyloid peptides (Abeta) has been investigated using hydrogen-deuterium exchange measured by electrospray ionization mass spectrometry and matrix-assisted laser desorption/ionization mass spectrometry. Mass spectrometric fragmentation of deuterated Abeta peptides was carried out by collision-induced dissociation, inlet fragmentation, and post-source decay. In contrast to the C-terminally truncated peptides Abeta(1-40) and Abeta(1-36) showing full hydrogen-deuterium exchange, Abeta(1-42) and the pyroglutamyl peptide Pyr(3)-Abeta(3-42) produced more complex signal patterns resulting from the formation of beta-sheet structured oligomers having 18-20 strongly protected protons. Using mass spectrometric fragmentation the results show that the reduced isotope exchange of Abeta(1-42) can be attributed to the central part of the chain comprising residues 8-23. This confirms involvement of the hydrophobic binding domain LVFFA in the course of Abeta aggregation and demonstrates that hydrogen-deuterium exchange in combination with mass spectrometry is well suited for structural analysis of monomeric and reversibly associated amyloid peptides using picomole quantities of material. PMID- 12539189 TI - Mass calibration of a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer including the rise time of the delayed extraction pulse. AB - To utilize fully modern MALDI-TOF and TOF/TOF mass spectrometers with mass resolution exceeding 10,000 and 2 ppm precision of flight time measurements for high mass accuracy, the model of ion motion used in the mass calibration equation must be expanded. The standard three-term equation providing up to 5-10 ppm (rms) mass accuracy with internal standards was modified with an additional term accounting for the finite rise time of the high-voltage extraction pulse. This new four-term calibration equation minimizes the effect of systematic error resulting from the fact that ion velocities are mass dependent due to the rise time of the extraction pulse. Applying this new calibration equation to a mass spectrum obtained in an axial MALDI-TOF MS containing 70 peaks (sodiated PEG), each with a signal-to-noise ratio greater than 100, a mass accuracy of 1.6 ppm (rms) was obtained over the mass range 1.0-4.0 kDa compared with 3.6 ppm (rms) with the standard three-term equation. The physical basis of the effects of the finite extraction pulse rise time on mass calibration is examined for axial MALDI TOF mass spectrometers, as well as for orthogonal acceleration TOF mass spectrometers. PMID- 12539190 TI - Liquid chromatography/mass spectrometry of malonyl-ginsenosides in the authentication of ginseng. AB - Different negative ion electrospray (ES) source conditions are required to concentrate the ion current in [M-H](-) for malonylated and non-malonylated ginsenosides. However, both can be ionised optimally in a single liquid chromatography/mass spectrometry (LC/MS) analysis by employing switchable voltages in the post-source ion optics of a quadrupole ion trap mass spectrometer. Coupled with automatic MS/MS scanning and post-acquisition neutral loss data analysis, this method provides a means of profiling the malonylated and acetylated ginsenosides in ginseng extracts. Analyses revealed numerous malonylated ginsenosides that could be partially characterised by serial MS/MS experiments. The ratio of mRb(1) to other isomeric forms present and to mRb(2) and mRc appears to show consistent differences among Panax ginseng (Asian ginseng), P. quinquefolius (American ginseng) and P. notoginseng (Sanchi ginseng). The ratio of malonylated to non-malonylated ginsenosides is reduced in the red form of Asian ginseng compared with the white form and there is a concomitant increase in the levels of the corresponding acetylated ginsenosides. The ability to analyse malonylated ginsenosides is an important contribution to the range of chemical characteristics that can be used to authenticate the different species of ginseng and will assist in quality control and standardisation. PMID- 12539191 TI - Ion suppression in the determination of clenbuterol in urine by solid-phase extraction atmospheric pressure chemical ionisation ion-trap mass spectrometry. AB - Ion suppression effects were observed during the determination of clenbuterol in urine with solid-phase extraction/multiple-stage ion-trap mass spectrometry (SPE/MS(3)), despite the use of atmospheric pressure chemical ionisation. During SPE, a polymeric stationary phase (polydivinylbenzene) was applied. Post cartridge infusion of analyte to the SPE eluate after the extraction of blank urine was performed to obtain a profile of the suppression. Single and multiple stage MS were performed to provide insight in the suppressing compounds. The ion suppression was mainly ascribed to two m/z values, but still no identification of the compounds was achieved from the multiple-stage MS data. No ionisable and non ionisable complexes and/or precipitation of clenbuterol with matrix compounds were observed. A concentration dependence of the percentage of suppression was observed. Up to 70% of the signal was suppressed upon post-cartridge infusion of 0.22 microg/mL (at 5 microL/min) clenbuterol into the eluate, and this decreased to about 4% at infusion of 22 microg/mL clenbuterol. Molecularly imprinted polymers were used to enhance the selectivity of the extraction. Although matrix components were still present after extraction, no interference of these compounds with the analyte was observed. However, the bleeding of the imprint from the polymer (brombuterol) caused significant ion suppression. PMID- 12539192 TI - Use of experimental design for optimisation of the cold plasma ICP-MS determination of lithium, aluminum and iron in soft drinks and alcoholic beverages. AB - A sensitive method for the simultaneous determination of (7)Li, (27)Al and (56)Fe by cold plasma ICP-MS was developed and validated. Experimental design was used to investigate the effects of torch position, torch power, lens 2 voltage, and coolant flow. Regression models and desirability functions were applied to find the experimental conditions providing the highest global sensitivity in a multi elemental analysis. Validation was performed in terms of limits of detection (LOD), limits of quantitation (LOQ), linearity and precision. LODs were 1.4 and 159 ng L(-1) for (7)Li and (56)Fe, respectively; the highest LOD found being that for (27)Al (425 ng L(-1)). Linear ranges of 5 orders of magnitude for Li and 3 orders for Fe were statistically verified for each compound. Precision was evaluated by testing two concentration levels, and good results in terms of both intra-day repeatability and intermediate precision were obtained. RSD values lower than 4.8% at the lowest concentration level were calculated for intra-day repeatability. Commercially available soft drinks and alcoholic beverages contained in different packaging materials (TetraPack, polyethylene terephthalate (PET), commercial cans and glass) were analysed, and all the analytes were detected and quantitated. PMID- 12539193 TI - Detection of Escherichia coli using immunomagnetic separation and bacteriophage amplification coupled with matrix-assisted laser desorption/ionization time-of flight mass spectrometry. AB - The application of whole cell analysis by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has emerged as a valuable tool for rapidly identifying/detecting bacteria. This technique requires minimal sample preparation and is simple to perform, but is generally limited to purified samples of bacteria at concentrations greater than 1.0 x 10(6) cells/mL. In this paper, we describe a bacterial detection method that integrates immunomagnetic separation with bacteriophage amplification prior to MALDI-MS analysis. The developed method consists of three main stages: (1) isolation of a target bacterium by immunomagnetic separation; (2) infection of the immuno-captured bacterium with a lytic bacteriophage; and (3) assay of infected medium for bacteriophage progeny using MALDI-MS to produce a molecular weight signal for the virus capsid protein. With this technique, the presence of Escherichia coli in broth was determined in less then 2 h total analysis time at a concentration of approximately 5.0 x 10(4) cells/mL. PMID- 12539196 TI - Subject-regulated dosing alters morphine self-administration behavior and morphine-stimulated [35S]GTPgammaS binding. AB - Repeated intake of opioids is associated with dose escalation and alterations in signal transduction at the G-protein-coupled receptor level. The current study utilized two experiments to identify factors in rats that influence consumption rates such as daily intake of self-administered morphine and receptor desensitization. In Experiment 1, rats self-administered either 0.30, 1.00, or 3.00 mg/kg/infusion morphine sulfate (morphine) during 7 daily 4-h sessions. For Experiment 2, rats were assigned to groups that self-administered either 1) self regulated escalating doses of morphine, 2) a fixed dose of morphine, or 3) saline during 18-h sessions for 7 days to determine if dose control would increase consumption without significantly decreasing response rate. We then assessed morphine-stimulated [(35)S]GTPgammaS binding in the amygdala and thalamus from these three groups in Experiment 2. Results from Experiment 1 demonstrated that 0.30 mg/kg/morphine did not support stable self-administration. For Experiment 2, the self-escalation group self-administered more morphine than the fixed-dose group, yet maintained similar response rates. Additionally, self-escalation rats demonstrated decreased morphine-stimulated [(35)S]GTPgammaS binding in membranes prepared from amygdalar and thalamic nuclei compared to the fixed-dose and control groups. Our results suggest that session length inversely affects consumption rates for fixed doses of morphine. Self-regulated dosing of morphine is also associated with rapid escalation of daily consumption and no significant alterations in consumption rates. These results suggest subject-regulated dosing is a useful approach for modeling dose escalation associated with opioid dependence. PMID- 12539197 TI - Link between dopamine D1 and D2 receptors in rat and human striatal tissues. AB - Many psychomotor behaviors depend on an interaction between dopamine D(1) and D(2) receptors. This study tested the hypothesis that agonist stimulation of dopamine D(1) receptors leads to the conversion of D(2) receptors from a state of high affinity for dopamine into a state of low affinity for dopamine. To test this hypothesis, dopamine was competed against [(3)H]raclopride for binding to rat and human striatal homogenates. Although the detection of high-affinity states of the dopamine D(2) receptor in rat or postmortem human striatum is normally difficult because the proportions of such sites are very low in the presence of physiological concentrations of sodium ions, the present work found that in the presence of 100 nM SCH 23390 to block D(1) receptors, a significant proportion of D(2) high-affinity sites were unmasked and readily revealed to be 10-35% of the D(2) population, illustrating the presence of a strong D(1)-D(2) link in both rat and human striata. PMID- 12539198 TI - Preproenkephalin mRNA expression in rat dorsal striatum induced by selective activation of metabotropic glutamate receptor subtype-5. AB - Group I metabotropic glutamate receptors (mGluR1 and mGluR5 subtypes) are positively coupled to phosphoinositide hydrolysis through G-proteins and are densely expressed in medium-sized projection neurons of striatum. Selective activation of Group I mGluRs upregulates preproenkephalin (PPE) mRNA expression in the rat dorsal striatum. This study investigated the role of one subtype of Group I receptors, mGluR5, in the regulation of PPE mRNA expression in the rat dorsal striatum using quantitative in situ hybridization. Unilateral injection of the mGluR5 selective agonist (RS)-2-Chloro-5-hydroxyphenylglycine (CHPG) into the dorsal striatum (caudoputamen) of chronically cannulated rats at doses of 50 and 200 nmol elevated basal levels of PPE mRNA in the injected dorsal striatum. The induction of PPE mRNA was evident at 1 h, remained at 3 h, and returned to normal level 6 h after CHPG injection. Pretreatment with an mGluR5 selective antagonist 2-methyl-6-(phenylethynyl) pyridine hydrochloride (MPEP) at a dose of 10 mg/kg (i.p.) blocked CHPG-stimulated PPE expression. MPEP also attenuated PPE expression induced by dopamine D(2) receptor blockade with eticlopride (0.5 mg/kg, i.p.). Administration of MPEP alone had no significant effects on basal levels of PPE mRNA in the striatum. The results from the present study demonstrate that glutamatergic tone on mGluR5 possesses the ability to positively regulate PPE gene expression in striatal neurons in vivo. Moreover, activation of mGluR5 participates in the mediation of D(2) antagonist-induced PPE expression. PMID- 12539199 TI - Relative levels of cytoprotection produced by analogs of choline and the role of alpha7-nicotinic acetylcholine receptors. AB - Several analogs of the acetylcholine precursor molecule choline have been widely studied as potential false cholinergic neurotransmitters with the therapeutic goal of using them to limit cholinergic neurotransmission. More recently, choline itself has been shown to act as a full, if low potency, agonist at the alpha7 subtype of the nicotinic acetylcholine receptor. This pharmacological property has been associated with the ability of nicotine and other related alpha7 receptor agonists to offer neuroprotection in a variety of experimental models. We confirm here that choline offers a significant degree of protection against the cytotoxicity induced by growth factor deprivation in differentiated PC-12 cells. Choline-induced cytoprotection ( approximately 1 mM) was about 3 orders of magnitude less potent than that for nicotine (EC(50) = 0.7 microM). Choline also exhibited only about 40% of the full cytoprotective effect of nicotine. Ethyl substitution for choline's N-methyl groups did not result in a significant improvement over choline as a cytoprotective agent. In contrast, pyrrolidinecholine exhibited much greater potency (EC(50) = 20 microM) and increased efficacy (about 55% of nicotine's effect) than choline. Like choline and nicotine, pyrrolidinecholine fully displaced [(125)I]alpha-bungarotoxin binding (K(i) = 33 microM) and chronic exposure to the analog increased cell surface binding sites. The cytoprotective effects of the analog were completely inhibited by coincubation with methyllycaconitine (MLA), a selective alpha7 nicotinic receptor antagonist. These findings are consistent with the possibility that the choline structure may serve as a template for the development of novel agents with both alpha7-nicotinic agonist activity and potential neuroprotective ability, as many of these compounds, including pyrrolidinecholine, are transported along with choline into the central nervous system. PMID- 12539200 TI - Neonatal ventral hippocampal damage modifies serum corticosterone and dopamine release responses to acute footshock in adult Sprague-Dawley rats. AB - Rats with excitotoxic neonatal ventral hippocampal lesions (NVHL) manifest in early adulthood a variety of behavioral and neurochemical abnormalities mimicking those seen in patients with schizophrenia. Some of these aberrations implicate malfunction of the midbrain dopamine systems. We studied NVHL effects on dopamine release in the rat frontal cortex, nucleus accumbens, and striatum during acute stress caused by inescapable continuous footshock (0.45 mA). Serum total corticosterone and prolactin levels were used as peripheral indices of stress. As an indirect index of dopamine release, tissue 3-methoxytyramine levels attained in vivo 10 min after monoamine oxidase inhibition was assayed in rats sacrificed by instantaneous microwave fixation of the brain tissue. Nonshocked NVHL rats showed significantly less nucleus accumbens' 3-methoxytyramine accumulation than their sham counterparts. Frontal cortical 3-methoxytyramine levels rose similarly after 20-min footshock in both groups of rats, but while it normalized after 60 min footshock in the sham rats, it did not decrease in the NVHL rats. Nucleus accumbens' 3-methoxytyramine was significantly elevated after either 20-min or 60 min footshock in both groups, whereas striatal 3-methoxytyramine was significantly elevated in the NVHL rats only. Serum corticosterone showed similar elevations in the sham and NVHL rats, but the patterns differed in that there was no attenuation after 60-min footshock in the latter. The lesion did not affect serum prolactin response. These data indicate that neonatal ventral hippocampal damage enhances and prolongs certain neural and neuroendocrine responses to acute physical stressor(s), and thus may affect adaptation and enhance detrimental effects of stress. PMID- 12539201 TI - Down-regulation of mGluR8 in pilocarpine epileptic rats. AB - Activation of presynaptic metabotropic glutamate receptors (mGluRs) leads to a powerful inhibition of glutamate release from many synaptic terminals throughout the CNS. mGluRs as autoreceptors are believed to provide a negative feedback system that prevents potentially toxic accumulation of glutamate in the extracellular space during synchronous synaptic activity such as epileptic seizures. In this study we analyzed the function of presynaptic mGluR8 on terminals of the lateral perforant pathway in the pilocarpine model of limbic epilepsy. Field excitatory postsynaptic potentials (fEPSPs) recorded in hippocampal slices of rats that developed spontaneous recurrent seizures after pilocarpine-induced status epilepticus (SRS group) showed a significantly reduced sensitivity to Group III mGluR agonists and severe mossy fiber sprouting. The Group III mGluR agonist L(+)-2-amino-4-phosphonobutyric acid (L-AP4, 10 microM) depressed fEPSPs in the SRS group only by 26 +/- 21% compared to 50 +/- 18% in untreated rats. Similarly, the mGluR8 preferring agonist (R,S)-4 phosphonophenylglycine (PPG, 5 microM) was significantly less effective in slices from SRS rats (43 +/- 4% vs. 83 +/- 5%). Concentration-response curves for L-AP4 revealed that the EC(50) values were not different between the control and SRS group (13 +/- 7 microM vs. 9 +/- 9 microM), while the maximal depressing effect was significantly reduced. The remaining depressing effect of L-AP4 in the SRS group could be blocked by the Group III specific antagonists (RS)-alpha-methyl-4 phosphonophenylglycine (MPPG) and alpha-methyl-L-AP4 (MAP4). Rats that did not develop SRS following pilocarpine-induced status epilepticus were indistinguishable from control rats: fEPSPs were highly sensitive to L-AP4 and there was no mossy fiber sprouting. The results show that pilocarpine-induced status epilepticus can lead to a downregulation of mGluR8 and suggest that the condition of SRS is associated with a deteriorated autoregulation of glutamate release. PMID- 12539202 TI - Calcium/calmodulin-dependent kinase II is involved in the facilitating effect of clozapine on NMDA- and electrically evoked responses in the medial prefrontal cortical pyramidal cells. AB - Using the method of intracellular recording in in vitro brain slices, we investigated whether calcium/calmodulin-dependent kinase II (CaMKII) is involved in the facilitating action produced by the atypical antipsychotic drug (APD) clozapine on N-methyl-D-aspartate (NMDA)-induced inward currents and electrically evoked excitatory postsynaptic currents (EPSCs) in pyramidal cells of the medial prefrontal cortex (mPFC). The CaMKII inhibitor, KN-93 (N-[2-(N-(4-Chlorocinnamyl) N-methylaminomethyl)phenyl]-N-[2-hydroxyethyl]-4-methoxybenzenesulfonamide), but not the inactive isomer, KN-92 (2-[N-(4-Methoxybenzenesulfonyl)]amino-N-(4 chlorocinnamyl)-N-methylbenzylamine, phosphate), blocked clozapine's augmenting effect on NMDA-evoked responses in pyramidal cells of the rat mPFC. KN-93 also inhibited the facilitatory effect of clozapine on electrically evoked responses in the pyramidal cells, while KN-92 did not show any effect. Similarly, the calmodulin antagonist W-7 (N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide) inhibited the augmenting effect of clozapine on NMDA- and electrically evoked responses in the pyramidal cells. To further test the role of CaMKII in mediating the augmenting action of clozapine, we performed experiments in alpha-CaMKII mutant and wild-type mice. In contrast to results in pyramidal cells from rats or wild-type mice, clozapine was not able to potentiate NMDA-induced currents in the mPFC pyramidal cells from the CaMKII mutant mouse. Both KN-93 and W-7, but not KN 92, inhibited the augmenting action of clozapine in the pyramidal cells of wild type mice. Taken together, these results suggest that the facilitating action of clozapine on the NMDA- and electrically evoked responses in pyramidal cells of the mPFC requires activation of CaMKII enzyme. PMID- 12539204 TI - Pathogenic role of glial cells in Parkinson's disease. AB - Parkinson's disease (PD) is a common neurodegenerative disorder characterized by the progressive loss of the dopaminergic neurons in the substantia nigra pars compacta (SNpc). The loss of these neurons is associated with a glial response composed mainly of activated microglial cells and, to a lesser extent, of reactive astrocytes. This glial response may be the source of trophic factors and can protect against reactive oxygen species and glutamate. Alternatively, this glial response can also mediate a variety of deleterious events related to the production of pro-oxidant reactive species, and pro-inflammatory prostaglandin and cytokines. We discuss the potential protective and deleterious effects of glial cells in the SNpc of PD and examine how those factors may contribute to the pathogenesis of this disease. PMID- 12539205 TI - S18Y polymorphism in the UCH-L1 gene and Parkinson's disease: evidence for an age dependent relationship. AB - We studied the relationship between Parkinson's disease (PD) and the S18Y polymorphism in the UCH-L1 gene and the effect on this relationship of age at onset, smoking, and pesticides. Patients requested free health coverage for PD to the Mutualite Sociale Agricole (MSA), the French health insurance organization for people whose work is related to agriculture. Controls requested reimbursement of health expenses to the MSA. A maximum of three controls were matched to each case. Analyses included participants with both parents born in Europe. There were no differences in S18Y genotypes between patients (n = 209; 67% SS, 32% SY, 1% YY) and controls (n = 488; 66% SS, 30% SY, 4% YY). The relationship between PD and S18Y was modified by age at onset (P = 0.03). The Y allele was inversely associated with PD for patients with onset before 61 years (odds ratio [OR] = 0.53; 95% confidence interval [CI], 0.29-0.99); there was no association for older patients (62-68 years: OR = 1.21; 95% CI, 0.67-2.20; >68 years: OR = 1.24; 95% CI, 0.67-2.31). Among patients, Y carriers had a later onset than noncarriers (P = 0.04). These findings were not modified or confounded by smoking and pesticides. In this community-based case-control study, carriers of the Y allele were at decreased risk of developing PD at a young age, independently of pesticides and smoking. PMID- 12539206 TI - Effects of CB1 cannabinoid receptor modulating compounds on the hyperkinesia induced by high-dose levodopa in the reserpine-treated rat model of Parkinson's disease. AB - The present study was designed to determine the potential of CB1 cannabinoid receptor modulating compounds in the treatment of L-3,4-dihydroxyphenylalanine (L dopa)-induced dyskinesia in Parkinson's disease. In the reserpine-treated rat model of parkinsonism, administration of a high dose of L-dopa (150 mg/kg) but not of Cl-APB (0.5 mg/kg) or quinpirole (0.5 mg/kg) produced a hyperkinetic state characterised by an increase in horizontal and vertical activity, which likely represent correlates of antiparkinsonian and dyskinetic activity, respectively. Injection of the CB1 cannabinoid receptor antagonist SR141716 (0.1-3 mg/kg) reduced the increase in vertical activity elicited by L-dopa without affecting the increase in horizontal activity. Injection of the CB1 cannabinoid receptor agonist WIN55,212-2 (0.1-3 mg/kg) reduced the L-dopa-induced increase in vertical activity and, at the highest dose only (3 mg/kg), also reduced horizontal activity elicited by L-dopa. WIN55,212-2 (1 mg/kg) reduced motor activity induced by both the D1 receptor agonist Cl-APB (0.5 mg/kg) and the D2 receptor agonist quinpirole (0.5 mg/kg) in the reserpine-treated rat. SR141716 (1 mg/kg) had no effects on motor activity induced by Cl-APB (0.5 mg/kg) nor quinpirole (0.5 mg/kg) in the reserpine-treated rat. Injection of the inhibitor of endocannabinoid transport AM404 (0.1-1 mg/kg) did not affect the increase in horizontal or vertical activity elicited by L-dopa (150 mg/kg) in the reserpine treated rat. The data suggest that both CB1 cannabinoid receptor antagonists and agonists can modulate the behavioural effects of L-dopa and may be useful for the treatment of the dyskinesia associated with long-term L-dopa treatment of Parkinson's disease. PMID- 12539207 TI - Deficit of verb generation in nondemented patients with Parkinson's disease. AB - Clinical and neuroimaging studies have shown that verb processing suggests a preferential participation of a prefrontal network, which is dysfunctional in Parkinson's disease (PD). To assess a verb processing deficit in PD, we compared noun- and verb-generation tasks for 34 nondemented PD patients (according to the Dementia Rating Scale) with 34 matched normal subjects, using two intracategory tasks (noun/noun and verb/verb generation) and two intercategory tasks (noun/verb and verb/noun generation). PD patients were significantly impaired in the two tasks involving verb production, i.e., verb/verb and noun/verb generation, whereas their performance was similar to those of controls in the two tasks requiring noun production. For the two impaired tasks, we assessed 1) the influence of lexical competition that corresponds to the presence of several candidate words for a given stimulus; 2) the influence of slight cognitive dysfunction; and 3) the influence of motor deficit. Significant correlations were found between DRS scores and performance on the noun/verb task, and no significant correlations were found between lexical competition or motor deficit and performance. The specific deficit for verb production in PD patients is discussed in relation to deficits affecting either action or grammatical representations. PMID- 12539209 TI - Long-term efficacy of thalamic deep brain stimulation for tremor: double-blind assessments. AB - Thalamic deep brain stimulation (DBS) is proven to suppress tremor in Parkinson's disease (PD) and essential tremor (ET). However, there are few reports on its long-term efficacy. We studied the efficacy of DBS at 2 years and 6-7 years after electrode implantations in the ventrointermediate nucleus of the thalamus in 39 patients (20 PD, 19 ET) with severe tremor. Twenty-five of the patients completed the study. Evaluations were done in a double-blind manner with the Unified Parkinson's Disease Rating Scale (UPDRS) and Essential Tremor Rating Scale (ETRS). DBS decreased tremor sum scores in PD (P < 0.025) compared to the preoperative baseline (median, 7; Q25-75, 6-9) both at 2 years (median, 2; Q25 75, 2-3.5; n = 16) and at 6 to 7 years (median, 2.5; Q25-75, 0.5-3; n = 12). Stimulation on improved tremor sum as well as sub scores (P < 0.025) compared to stimulation off conditions. In ET, thalamic stimulation improved (P < 0.025) kinetic and positional tremor at both follow-up periods (n = 18 and n = 13, respectively) with significant improvements (P < 0.025) in hand-function tests. PD but not ET patients showed a general disease progression. Stimulation parameters were remarkably stable over time. We conclude that high-frequency electric thalamic stimulation can efficiently suppress severe tremor in PD and ET more than 6 years after permanent implantation of brain electrodes. PMID- 12539208 TI - Leg muscle strength is reduced in Parkinson's disease and relates to the ability to rise from a chair. AB - Individuals with Parkinson's disease (PD) have difficulties rising from a chair; however, factors contributing to this inability have never been investigated. We compared lower extremity strength between individuals with PD and healthy controls and quantified the relationships between strength and the ability to rise from a chair. Ten men with mild PD and 10 sex- and age-matched controls performed maximal concentric, isokinetic knee and hip extensor torque on an isokinetic dynamometer to quantify muscle strength. Subjects also rose from a chair at their comfortable pace without the use of their arms and the duration of this task provided a measure of sit-to-stand (STS) ability. Subjects with PD were tested in an on- and off-medication state on different days. Mean hip and knee extensor torques were lower in subjects with PD, with greater deficits found at the hip. Greater hip strength was related to better STS ability in subjects with PD while greater knee strength was related to better STS ability in controls. These results show that individuals with mild PD generate smaller extremity forces compared to controls. Reduced strength, particularly at the hip, may be one factor that contributes to the difficulty of persons with PD to rise from a chair. PMID- 12539210 TI - Impact of social functioning and vitality on preference for life in patients with Parkinson's disease. AB - The determinants of preference for life in patients with Parkinson's disease are not well known. We assessed the effect of functional status on the preference for life as measured by the time trade-off method with a 10-year life span. Our survey was based on a random sample of 1,200 patients from the Japanese Association of Patients with Parkinson's Disease. Patients' demographics, clinical information, and functional status as measured by the MOS Short Form 36 were considered independent variables. The response rate was 63.5%. Linear regression showed that men had a significantly stronger preference for current health than women (by 10.4 months on a scale of 10 years). Patients with higher physical functioning, social functioning, and vitality had significantly higher preferences for life (each 10-point improvement in physical or social functioning led to a 1.5-month increment in preference for current health; a 10-point improvement in vitality led to a 3-month increment). Longer duration of disease and advanced Hoehn and Yahr stage were significantly associated with a lower preference for current health (by 0.5 months/year of disease and by 2.6 months/stage). Interventions that target social functioning and vitality may be beneficial to preference for life. PMID- 12539211 TI - Double-blind, single-dose, cross-over study of the effects of pramipexole, pergolide, and placebo on rest tremor and UPDRS part III in Parkinson's disease. AB - Tremor is one of the cardinal signs of Parkinson's disease (PD) but its response to antiparkinsonian medication is variable. It has been postulated that pramipexole may have a stronger antiparkinsonian tremor effect than pergolide, another direct acting dopamine agonist medication, possibly because the former has preferential affinity for the dopamine D3 receptor. The purpose of this pilot study was to compare the effects of a single oral dose of either pramipexole (Pr) or pergolide (Pe) or placebo (Pl) on parkinsonian tremor and the motor (part III) subsection of the UPDRS. Ten patients (6 men, 4 women), mean age 65.3 years, mean duration from diagnosis of 2.6 years, with tremor dominant PD were recruited. On three separate occasions a single dose of pramipexole (salt) 500 microg, pergolide 500 microg or placebo were administered in random order to each patient, who were pretreated with domperidone and had their antiparkinsonian medication withheld from midnight before study. After each medication patients were assessed at baseline and then every 30 min for 4 hr using a 0 to 10 tremor rating scale and the UPDRS (part III) in a double-blind protocol. Adverse effects were systematically recorded. The results demonstrate that 500 microg of either pramipexole or pergolide reduced PD rest tremor scores to a similar degree, which at peak effect was significantly greater than placebo (respectively Pe v Pl: P < 0.006, Pr v Pl: P < 0.033). The two active drugs also had weaker beneficial effects on the UPDRS part III. Pergolide, however, was significantly more likely than pramipexole to cause nausea (P = 0.005) or vomiting (P = 0.014). PMID- 12539212 TI - Restless legs syndrome in Parkinson's disease: a case-controlled study. AB - Restless legs syndrome (RLS) is a disorder of motor activity with a circadian pattern, occurring frequently in patients with Parkinson's disease (PD). We sought to estimate the prevalence of RLS in Indian PD patients. One hundred twenty-six consecutive PD patients and 128 healthy age- and sex-matched controls were evaluated using a predesigned questionnaire. RLS was present in 10 of 126 cases of PD (7.9%) and 1 of 128 controls (0.8%, P = 0.01). PD patients with RLS were older than those without RLS (63.70 +/- 7.80 years vs. 57.37 +/- 10.04 years; P = 0.05) and had higher prevalence of depression (40% vs. 10.3%; P = 0.023). No demographic factors or factors related to PD correlated with the presence or severity of RLS. RLS is more common among patients with PD than controls. A greater medical recognition of this disorder is needed in view of available effective treatment. PMID- 12539213 TI - Cerebrospinal fluid Abeta42 is reduced in multiple system atrophy but normal in Parkinson's disease and progressive supranuclear palsy. AB - The 42-amino-acid isoform of beta-amyloid Abeta42 in the cerebrospinal fluid (CSF) has recently been proposed as a biochemical marker for Alzheimer's disease (AD) and subcortical white-matter dementia (SWD). In both of these conditions, concentration of CSF-Abeta42 is reduced. We quantified CSF-Abeta42 from patients fulfilling strict clinical criteria for multiple system atrophy (MSA; n = 36), Parkinson's disease (PD; n = 48) and progressive supranuclear palsy (PSP; n = 15). The study groups were consecutively recruited among patients referred to a movement disorder unit, and 32 healthy, age-matched volunteers were used as controls. The CSF concentration of Abeta42 was significantly reduced in the MSA group (P < 0.001), whereas the PD and PSP groups did not differ from controls. On an individual basis, low content of Abeta42 was seen in 9 MSA patients regardless of age and disease duration. Three PD patients with long disease duration also had low concentrations but all PSP patients were normal. We conclude that the reduced CSF-Abeta42 concentration may be a clue to the pathogenesis of MSA. There is a decreased production, or more possible, an increased consumption of CSF Abeta42. The analysis of this protein may also become a supplement to the clinical differentiation of parkinsonian syndromes in a movement disorder unit. PMID- 12539214 TI - Acute effect of transcutaneous electrical nerve stimulation on tremor. AB - Based on the claims that transcutaneous electrical nerve stimulation is effective in myoclonic dystonia and essential tremor, we evaluated its acute effects in 5 patients with essential tremor and 2 patients with tremor attributed to peripheral neuropathy using as parameters the Washington Heights-Inwood Genetic Study of Essential Tremor rating scale, self-reported impression, and recording of electromyographic activity. We found no significant improvement in any of the parameters tested. PMID- 12539215 TI - Primary orthostatic tremor is an exaggeration of a physiological response to instability. AB - Primary orthostatic tremor (POT) is a rare disorder characterised by an intense sense of unsteadiness upon standing and a 16-Hz tremor in which the timing between tremor bursts in different muscles (unilateral and bilateral) remains constant. Hitherto, similar EMG activity has not been described in healthy subjects and it has been postulated that the oscillations seen in POT are primarily pathological. In this study, EMG was recorded from tibialis anterior in healthy subjects who were made unsteady through vestibular galvanic stimulation or leaning backwards. Under these conditions, a peak at approximately 16 Hz was seen in the coherence between the left and right tibialis anterior. This bilateral coherence was absent when the subjects activated the same muscles when not unsteady. These data indicate the existence of a physiological system involved in organising postural responses under circumstances of imbalance and characterised by a highly synchronised output at approximately 16 Hz. In addition, the results suggest that the core abnormality in POT may be an exaggerated sense of unsteadiness when standing still, which then elicits activity from a 16-Hz oscillator normally engaged in postural responses. PMID- 12539216 TI - Slowly progressive cerebellar ataxia and cervical dystonia: clinical presentation of a new form of spinocerebellar ataxia? AB - We describe 5 cases with a rare combination of young-onset, slowly progressive cerebellar ataxia and cervical dystonia. Two were sporadic, whereas the other 3 were familial, including 2 from one family. The age of onset of these cases was between 16 and 37 years. The presenting symptom was cervical dystonia and/or dystonic head tremor in 3 patients and hand or lower limb tremor in 2. In 2 cases, cervical dystonia and/or dystonic head tremor developed approximately 6 to 10 years before cerebellar dysfunction, and in three they developed at the same time. Apart from cervical dystonia, there was mild dystonic limb involvement in 2 cases, but generalized dystonia was not seen. Cerebellar ataxia was slowly progressive. A literature search showed 10 cases of cervical dystonia associated with genetically undetermined (n = 5) or genetically proven (n = 5) spinocerebellar ataxia (SCA). When the genotype was known, these patients had either SCA3, 6, 7, or 12. However, our 5 cases (or their first-degree relatives) tested negative for SCA1, 2, 3, 6, and 7, and in the 4 cases (or their first degree relatives) tested for SCA12, the result was negative. We propose that this rare phenotype manifesting as a combination of cerebellar ataxia and cervical dystonia may represent one or more new, as yet uncharacterized, genotypes of inherited young-onset spinocerebellar ataxia. PMID- 12539217 TI - Familial blepharospasm is inherited as an autosomal dominant trait and relates to a novel unassigned gene. AB - Blepharospasm (BSP) is a common form of primary torsion dystonia (PTD). Although most cases are sporadic, an increased familial incidence of BSP has been reported. Precisely how blepharospasm is inherited remains unclear. We report on two Italian families with adult-onset focal BSP inherited as an autosomal dominant trait with reduced penetrance. None of the affected family members had the 3-bp (GAG) or the 18-bp deletion in the DYT1 gene. In one family, linkage analysis allowed us to exclude segregation of the disease with the known PTD loci (DYT1, DYT6, DYT7, and DYT13). These findings suggest that primary familial adult onset BSP is a distinct entity among inherited PTD and is caused by a novel, unmapped gene. PMID- 12539218 TI - Substantia nigra lesions in viral encephalitis. AB - Post-encephalitic parkinsonism is a well-known entity, but involvement of the basal ganglia is rarely documented. We describe a 21-year-old man who developed parkinsonism following encephalitic illness presumed to be of viral origin with substantia nigra lesions evident on magnetic resonance imaging scan. PMID- 12539219 TI - Secondary progressive chronic manganism associated with markedly decreased striatal D2 receptor density. AB - We describe a patient with chronic manganism due to intoxication 40 years ago. Whereas previous reports on acute or subacute intoxication have shown no or only small reductions in striatal D2 receptor density, we found markedly decreased D2 receptor density using (18)F-methylspiperone PET in this very late stage of chronic manganism, supporting the hypothesis that manganese intoxication may trigger a neurodegenerative disease process. PMID- 12539220 TI - Dopaminergic response in Parkinsonian phenotype of Machado-Joseph disease. AB - We report on a patient with genetically proven Machado-Joseph Disease (MJD) presenting with signs indistinguishable from Parkinson's disease (PD), including levodopa response and typical levodopa-induced motor fluctuations. Only after 10 years of prolonged benefit from levodopa and different dopamine agonists (DA), the patient developed cerebellar ataxia and pyramidal signs. Preferential D3 receptor-stimulating dopamine agonists especially showed a benefit at the time, when D2 receptor binding was reduced in IBZM SPECT. This is the first report of a meaningful response to DA in MJD. PMID- 12539221 TI - Chronic thalamic stimulation in a patient with spinocerebellar ataxia type 2. AB - We report on a patient with spinocerebellar ataxia type 2 (SCA 2) with an unusual clinical presentation, including severe, disabling resting and action tremor and the successful treatment of this tremor syndrome with chronic thalamic stimulation. Using [(123)I]beta-CIT single photon emission computed tomography, we document a marked degeneration of the nigrostriatal dopaminergic system in SCA 2. PMID- 12539222 TI - Genetic association of Huntington's disease and restless legs syndrome? A family report. AB - We report on a family with comorbidity of Huntington's disease (HD) and idiopathic restless legs syndrome (RLS). All family members investigated and affected by RLS are also affected by HD, but not vice versa. The association of HD and RLS is discussed with respect to dopaminergic dysfunction. PMID- 12539223 TI - Dissociation between androgen responsiveness for malignant growth vs. expression of prostate specific differentiation markers PSA, hK2, and PSMA in human prostate cancer models. AB - BACKGROUND: A detailed understanding is evolving as to how androgen receptor (AR) functions as a transcriptional regulator via its binding to androgen response elements (ARE) within promoter and enhancer regions of prostate-specific differentiation markers such as PSA, hK2, and PSMA. It has been assumed that an understanding of regulation of expression of these marker proteins would also provide an understanding of the mechanisms whereby AR interactions regulate proliferation and survival of malignant prostate cells. In order to validate this hypothesis, we used a series of human prostate cancer models [i.e., LAPC-4, CWR22Rv1, MDA PCA-2b, LNCaP, and C4-2B (derived from LNCaP)] to test whether there is a consistent concordance between androgen responsive regulation for malignant growth vs. regulation of expression of prostate differentiation specific markers PSA, hK2, and PSMA. METHODS: In order to define androgen growth responsiveness in vivo, human prostate cancer cell lines were inoculated as xenografts into intact vs. surgically castrated adult male nude mice and the subsequent tumor growth response monitored. To assess androgen regulation of PSA and hK2 expression in these cell lines, the concentration of PSA and hK2 in the conditioned standard media and charcoal stripped media +/- androgen from each cell line was determined using an immunoassay system. PSMA enzymatic activity was determined using the PSMA substrate (3)H N-acetylaspartylglutamate ((3)H NAAG). RESULTS: Wild-type AR expressing LAPC-4 cells are androgen responsive for their in vivo growth. This cell line is also androgen sensitive for the expression of both PSA and hK2 in vitro and express PSMA. CWR22Rv1 cells have a mutated AR and are androgen responsive for growth in vivo and androgen sensitive for hk2 but not PSA expression. CWR22Rv1 produce approximately 1.4-fold more PSA, approximately 18-fold more hK2, and have 21-fold higher PSMA activity than LAPC-4 cells. MDA PCA-2b cells are androgen responsive for growth in vivo and androgen sensitive for PSA expression. MDA PCA-2b cells produce approximately 250-fold more PSA but almost equivalent amounts of hK2 compared to LAPC-4 and have approximately 19 fold higher PSMA activity. Both late passage LNCaP and C4-2B are androgen independent for growth in vivo but remain androgen sensitive for both PSA and hK2 expression. LNCaP cells produce approximately 50-fold more PSA, approximately 35 fold more hK2, and have 28-fold higher PSMA activity compared to LAPC-4. C4-2B cells produce approximately 80-fold higher levels of PSA, approximately 250-fold higher levels of hK2. C4-2B also the highest PSMA activity of the cell lines with 105-fold higher PSMA activity than LAPC-4 and approximately 4-fold higher activity than late passage LNCaP cells. CONCLUSIONS: Androgen can coordinately regulate both the tumor growth and expression of prostate specific marker genes as observed for the LAPC-4 human prostate cancer cells. Such coordinated regulation, however, is not universal. In all of the other cell lines, there is a dissociation between androgen responsive regulation of malignant growth vs. regulation of expression of prostate specific markers PSA and hK2. In addition, PSMA activity in these cell lines increases as cells become more androgen independent for growth in vivo. These results emphasize that tumor growth and the expression of the specific secretory genes are independently regulated molecular events even if they share a requirement for androgen and/or AR function. Additional independent mechanisms occur in prostate cancer cells for regulation of expression for even the highly related PSA and hK2 genes. Further studies are needed to clarify the mechanisms for androgen ligand-independent, AR-dependent regulation of the genes that directly effect the growth of androgen (i.e., ligand) independent prostate cancer cells. Unfortunately, the data in this present report do not validate the use of the PSA or hK2 gene as surrogates for a model system for such critically important mechanistic studies. Prostate prostate cancer cells. Unfortunately, the data in this present report do not validate the use of the PSA or hK2 gene as surrogates for a model system for such critically important mechanistic studies. PMID- 12539224 TI - Leptomycin B stabilizes and activates p53 in primary prostatic epithelial cells and induces apoptosis in the LNCaP cell line. AB - BACKGROUND: Previous studies showed that primary cultures of normal and malignant human prostatic epithelial cells are defective in their ability to upregulate the tumor suppressor protein p53 in response to DNA damage. This dysfunctional regulation of p53 may be relevant to both the high incidence of prostate cancer and its resistance to chemotherapy. Leptomycin B (LMB) has recently been found to increase the protein level and transcriptional activity of p53 by interfering with nucleocytoplasmic export and subsequent degradation by the proteasome. We investigated the ability of LMB to activate p53 in prostatic epithelial cells. METHODS: Primary cultures and the cell lines LNCaP and DU 145 were treated with LMB. p53 protein was evaluated in Western blots and by immunocytochemistry. Induction of downstream targets of p53 was evaluated in Western and Northern blots. Growth inhibition, cell cycle arrest, and apoptosis in response to LMB were measured in clonal growth assays, by flow cytometry, and by Hoescht/propidium iodide staining, respectively. RESULTS: Treatment of prostatic epithelial cells with LMB led to post-translational stabilization of p53, activation of downstream target genes, and induction of cell cycle arrest in primary cultures and apoptosis in LNCaP (with wild-type p53) but not DU 145 (with mutant p53) cells. CONCLUSIONS: p53 in primary cultures of normal and malignant prostate cells, although dysfunctional in that it is not responsive to DNA damage, is activated by LMB. The ability of LMB to stabilize p53 and induce expression of p53-responsive growth inhibitory genes may be a useful lead in the development of chemopreventive or therapeutic small molecules that can modulate p53 function in prostatic epithelial cells. PMID- 12539225 TI - Increased expression of bone morphogenetic protein-7 in bone metastatic prostate cancer. AB - BACKGROUND: Prostate cancer frequently metastasizes to bone. The skeletal metastases of prostate cancer origin are osteoblastic rather than osteolytic. Recently, the expression of bone morphogenetic proteins (BMPs) in prostate cancer cell lines was detected. The present study indicated the existence of BMP-7 in normal prostate tissue, but its function has not been clarified. The mechanism by which prostate cancer causes osteoblastic metastasis is not clear. We investigated the expression of BMP-7 and -6 in normal and metastatic bone tissues to clarify the biological relationship between the expression of BMPs and bone metastasis in prostatic cancer. METHODS: Six samples of normal bone tissue and nine samples of bone metastasis tissue were collected during the autopsies of six patients with prostate cancer. Total RNA was extracted from these samples. After reverse transcription (RT) of the RNA samples, the expression of BMP-6 and -7 in each sample was measured by the real-time quantitative polymerase chain reaction (PCR). Glyceraldehyde phosphate dehydrogenase (GAPDH) was used as an internal standard. RESULTS: Although the expression of BMP-7 was detected in five out of seven (71%) metastatic bone lesions of prostate cancer, it was not detected in normal bone tissues. The expression level of BMP-7 was significantly higher in metastatic bone lesions than in normal bone (P < 0.05). There was no significant difference between the level of expression of BMP-6 in metastatic bone lesions from prostate cancer and the level in normal bone tissue (P = 0.81). CONCLUSIONS: These results suggest that high expression of BMP-7 in metastatic bone lesions of prostate cancer is related to osteoblastic metastasis. BMP-7 in the bone metastasis tissue indicates that the cells expressing BMP-7 probably originated from the prostate, because we have detected high expression of BMP-7 in the prostate. Prostate 54: 268-274, 2003. PMID- 12539226 TI - Selective reduction of AKR1C2 in prostate cancer and its role in DHT metabolism. AB - BACKGROUND: As androgens play an essential role in prostate cancer, we sought to develop a real-time PCR to characterize mRNA expression profiles of human members of the Aldo-Keto Reductase (AKR) 1C gene family, as well as of 5 alpha-steroid reductase Type II (SRD5A2) in prostate cancer samples. Functional activity and regulation of AKR1C2, a 3 alpha-hydroxysteroid dehydrogenase (HSD) type III, was also assessed in prostate cancer cell lines. METHODS: Gene specific PCR primers were established and relative gene expression of human AKR1C family members was determined in paired samples of cancerous and surrounding unaffected prostate tissue. RESULTS: AKR1C2 preferentially reduces DHT to the weak metabolite 5 alpha androstane-3 alpha,17 beta-diol (3 alpha-diol) without conversion of 3 alpha-diol to DHT in the PC-3 cell line, and its expression was increased by DHT treatment in LNCaP cells. Selectively reduced expression of AKR1C2 mRNA, but not AKR1C1 (97% sequence identity), was found in approximately half of the pairs whereas AKR1C3 relative expression was not significantly altered. No aberrant expression of AKR1C4 expression or significant differences in SRD5A2 gene expression were found. CONCLUSIONS: AKR1C2 functions as a DHT reductase in prostate-derived cells lines and is regulated by DHT. Additional studies are needed to further define the significance of reduced AKR1C2 expression in prostate cancer and its potential role in modulating local availability of DHT. PMID- 12539227 TI - Expression of cystatins, high molecular weight cytokeratin, and proliferation markers in prostatic adenocarcinoma and hyperplasia. AB - BACKGROUND: Prostatic adenocarcinoma is the most common malignancy among men in the western world but the diagnostic and prognostic criteria for it are still not clearly defined. Additional means for its diagnosis and prognosis are clearly needed. Previously it has been shown that cystatin A is expressed in the basal cells of normal prostate and the expression disappears in prostatic carcinoma. METHODS: We have now studied the expression of both cystatins A and B in benign prostatic hyperplasias (BPH), prostatic intraepithelial neoplasias (PIN) and carcinomas of the prostatic epithelium and compared it with the expression of high molecular weight (HMW) cytokeratin as well as the proliferation markers cyclin A and Ki-67. The expression of the proteins was immunohistochemically assessed using 33 total prostatectomy specimens. RESULTS: Cystatin A was expressed in the basal cells in all cases of BPH, low-grade PIN, and high-grade PIN whereas carcinomas showed no staining of cystatin A. The 34 beta E12 cytokeratin expression was similar to basal cystatin A staining and was not seen in carcinoma foci. Cystatin B showed both nuclear and cytoplasmic expression in the columnar epithelial cells. The decrease in median cytoplasmic staining of cystatin B in carcinomas compared to other lesions was significant, but there was a significant increase in expression with dedifferentiation of carcinoma. Also cyclin A and Ki-67 staining were significantly different in non-carcinomatous foci compared to carcinoma foci and had a remarkably similar negative correlations with basal cystatin A and 34 beta E12 staining. CONCLUSIONS: The results show that cystatin expression can be used as an aid in the diagnosis of prostatic adenocarcinoma and especially cystatin A in the distinction between high grade PIN and grade I carcinoma. PMID- 12539228 TI - Molecular cloning and expression of a variant form of hippostasin/KLK11 in prostate. AB - BACKGROUND: Hippostasin is a kallikrein-like serine protease, which has two alternatively spliced isoforms, brain-type and prostate-type. We previously reported alternative expression of hippostasin in prostate cancer cell lines. METHODS: We studied the expression of a variant-form hippostasin (isoform 3) mRNA by RT-PCR. Localization of the isoform 3 protein was examined by immunohistochemistry. The enzymatic activity of the recombinant protein was measured with synthetic substrates. RESULTS: A novel isoform of hippostasin contains 25 additional amino acids in the catalytic triad of brain-type hippostasin. Its mRNA was expressed in normal prostate tissue, BPH, and prostate cancer cell lines. The protein was localized in the prostate secretory epithelium. The enzyme activity was similar to that of brain-type hippostasin, which has kallikrein-like activity. CONCLUSIONS: In this study, we have identified a third isoform of hippostasin, which was designated variant-form (isoform 3). Hippostasin isoform 3 may play a role in the prostate, including reproductive and/or tumorigenic functions. PMID- 12539229 TI - R726L androgen receptor mutation is uncommon in prostate cancer families in the united states. AB - BACKGROUND: A mutation in the androgen receptor (AR) gene, namely AR R726L, was described in 2% of Finnish men with sporadic or familial prostate cancer and was associated with an approximately sixfold increased risk of prostate cancer. We set out to determine the incidence of this mutation in a sample of men with either early-onset and/or familial prostate cancer in the United States. METHODS: Five hundred forty-eight men with prostate cancer from 411 unrelated families participating in the University of Michigan Prostate Cancer Genetics Project (PCGP) were studied. Allele-specific oligonucleotide hybridization was used to detect the presence of the AR R726L mutation in germline DNA. RESULTS: None of the 548 prostate cancer patients studied, including 513 White, 29 African American, 3 Asian, and 3 Hispanic men, were found to carry the AR R726L allele. Therefore, the prevalence of this allele is significantly less than that observed among Finnish men with prostate cancer (Fisher's exact test, P = 0.002). CONCLUSIONS: The AR R726L allele does not account for a significant proportion of early-onset and/or familial prostate cancer in the United States. PMID- 12539230 TI - Lesions missed on prostate biopsies in cases sent in for consultation. AB - BACKGROUND: There are no reports on how often lesions are missed on prostate needle biopsies. METHODS: Over a 10-month period, 8/99 to 5/00, 3,251 prostate biopsy cases were seen in consultation. RESULTS: We identified 87 (2.7%) patients with missed lesions (n = 9 academic hospitals; n = 44 community hospitals; n = 34 commercial labs). Overall, 119 lesions were missed in 87 patients. Missed lesions were as follows: small atypical glands suspicious for cancer (41 lesions in 35 patients), prostatic adenocarcinoma (39 cancers in 32 patients), high grade prostatic intraepithelial neoplasia (HGPIN) (34 lesions in 30 patients), and HGPIN with adjacent small atypical glands (five lesions in five patients)--some men with more than one type of missed lesion. Detection of the missed lesions would have resulted in either: a definite change in care in 15 of 3,251 (0.5%) patients or a possible change in care (bilateral cancer vs. unilateral cancer; HGPIN vs. atypical) in 17 (0.5%) patients. In 21 (24%) of the cases, the slides were seen by at least two pathologists prior to consultation at our hospital. CONCLUSIONS: Although the number of prostate biopsies with missed lesions in a consult-based population of prostate biopsies appears relatively high (2.7%), the detection of the missed lesions would have only effected a definite change in care in 0.5% of all patients or a possible change in care in another 0.5% of patients. Our data underestimates missed lesions, as the entire specimen was not submitted for review in 41% of cases. Although our incidence of missed lesions gives some indication as to the magnitude of the problem, it cannot be equated with the risk of missing lesions in unselected cases. PMID- 12539231 TI - Selenomethionine does not affect PSA secretion independent of its effect on LNCaP cell growth. AB - BACKGROUND: Individuals supplemented with selenium have reduced incidence of prostate cancer. This study determines whether selenomethionine specifically affects the secretion of prostate specific antigen (PSA) in vitro. METHODS: LNCaP cells were supplemented with selenomethionine for 7 days. PSA secretion was determined by ELISA. Cell proliferation was assessed by enumeration of trypan blue excluding cells. Colony formation was determined in soft agar. Cell cycle distribution was determined by FACS analysis of propidium iodide stained cells. RESULTS: Selenomethionine at > or = 70 microM inhibited LNCaP cell growth and colony formation. 0-100 microM selenomethionine did not affect the secretion of PSA by LNCaP cells in cell culture supernatants when normalized to the number of cells in culture. At supra-nutritional concentrations of selenomethionine, LNCaP cells had longer G(0)/G(1) phase in agreement with the inhibitory effects on cell growth. CONCLUSIONS: PSA secretion is not specifically inhibited by concentrations of selenomethionine corresponding to plasma selenium concentrations found in individuals supplemented with chemopreventive concentrations of selenized yeast. These data suggest that changes in serum PSA levels in individual patients during selenium supplementation is not an effect specific for PSA secretion, but rather may be a useful indicator for changes in disease progression in individual patients. PMID- 12539232 TI - Characteristics of human prostasomes isolated from three different sources. AB - BACKGROUND: Prostasomes are organelles secreted by human prostate epithelial cells and appear freely in semen. Most prostasomal functional and biochemical characteristics have been settled on basis of seminal prostasomes (SP). METHODS: Three types of prostasomes, viz., from seminal plasma, prostate gland (native), and prostate cancer bone metastasis, were isolated and compared for their biochemical and functional characteristics. RESULTS: Flow cytometry showed that all three types of prostasomes had a similar size distribution and electrophoresis revealed close similarities in banding pattern. Also, a high cholesterol/phospholipid ratio and some marker enzymes were distinctive common features. The unusual chromogranin B (CgB)/chromogranin A (CgA) ratio with CgB in abundance over CgA, previously found only in SP, was also valid for the native (NP) and metastasis-derived prostasomes (MP). The latter prostasome type contained a 10-fold higher amount of CgA compared to the others. SP and MP showed the same promotive effect on sperm forward motility, whereas NP did not. All three prostasome types exhibited an antibacterial activity against Bacillus megaterium. CONCLUSIONS: Since the three prostasome types were isolated from different environments, our data support the view that the common characteristics were prostasome-genuine. PMID- 12539233 TI - Sustainable development, biotechnology and the North-South divide. PMID- 12539234 TI - Detection of unpaired DNA at meiosis results in RNA-mediated silencing. AB - During meiosis, homologous chromosomes must pair in order to permit recombination and correct chromosome segregation to occur. Two recent papers show that meiotic pairing is also important for correct gene expression during meiosis. They describe data for the filamentous fungus Neurospora crassa that show that a lack of pairing generated by ectopic integration of genes can result in silencing of genes expressed during meiosis. This can result in aberrant meioses whose defects are specific to the function of the unpaired gene. Furthermore, mutations affecting the silencing mechanism have been selected in a gene encoding a putative RNA-dependent RNA polymerase. This finding indicates the involvement of a meiotic specific post-transcriptional gene silencing mechanism (PTGS) similar to that observed in vegetative cells in N. crassa and other organisms. Finally, this gene product is essential for normal meiosis, suggesting that RNA-dependent processes are fundamental to the sexual cycle. PMID- 12539235 TI - Cancer therapy: switching off oncogenes. AB - Cancer derives from a cell clone that has accumulated genetic and epigenetic changes that influence its phenotype and finally enable it to escape from the normal controls of proliferation. A recent paper shows that, in myc-induced tumours, the inactivation of this oncogene produces the regression of the tumours and the differentiation of the tumour cells into mature osteocytes.1 In addition, a further reactivation of myc in these cells does not restore the malignant phenotype but induces apoptosis. This discovery could lead to an innovative therapeutic strategy. PMID- 12539236 TI - Closing the (Ran)GAP on segregation distortion in Drosophila. AB - Segregation Distorter (SD) is a meiotic drive system in Drosophila that causes preferential transmission of the SD chromosome from SD/SD(+) males owing to induced dysfunction of SD(+) spermatids. Since its discovery in 1956, SD and its mode of action have baffled biologists. Recently, substantial progress has been made in elucidating this puzzle. Sd, the primary gene responsible for distortion encodes a mutant RanGAP, a key protein in the Ran signaling pathway required for nuclear transport and other nuclear functions. The mutant protein is enzymatically active but mislocalized to nuclei, which apparently disrupts Ran signaling by reducing intranuclear Ran-GTP levels. Some evidence suggests that a defect in nuclear transport may be the main cause of sperm dysfunction. Although important questions remain, the basic mechanism of distortion is now understood sufficiently well that specific hypotheses can be formulated and tested. This previously mysterious genetic system may now offer unique insights into novel aspects of regulation by Ran. PMID- 12539237 TI - Paradoxes of eukaryotic DNA replication: MCM proteins and the random completion problem. AB - Eukaryotic DNA replication initiates at multiple origins. In early fly and frog embryos, chromosomal replication is very rapid and initiates without sequence specificity. Despite this apparent randomness, the spacing of these numerous initiation sites must be sufficiently regular for the genome to be completely replicated on time. Studies in various eukaryotes have revealed that there is a strict temporal separation of origin "licensing" prior to S phase and origin activation during S phase. This may suggest that replicon size must be already established at the licensing stage. However, recent experiments suggest that a large excess of potential origins are assembled along chromatin during licensing. Thus, a regular replicon size may result from the selection of origins during S phase. We review single molecule analyses of origin activation and other experiments addressing this issue and their general significance for eukaryotic DNA replication. PMID- 12539238 TI - Stem cells, telomerase and dyskeratosis congenita. AB - Dyskeratosis congenita is a rare skin and bone marrow failure syndrome caused by defective telomere maintenance in stem cells. The major X-linked form of the disease is due to mutations in a nucleolar protein, dyskerin, that is part of small nucleolar ribonucleoprotein particles that are involved in processing ribosomal RNA. It is also found in the telomerase complex, pointing to an unexpected link between these two processes. An autosomal dominant form is due to mutations in the RNA component of telomerase (hTR). Patients with this form of the disease are more severely affected in later generations that carry the mutations, possibly due to the inheritance of shortened telomeres, disguising the inherited nature of the disease in some cases classified as aplastic anemia. Because of the importance of telomerase in tumour formation and aging, study of this disease may provide important clues about these fundamental processes. PMID- 12539239 TI - Molecular genetics of aging in the fly: is this the end of the beginning? AB - How we age and what we can do about it have been uppermost in human thought since antiquity. The many false starts have frustrated experimentalists and theoretical arguments pronouncing the inevitability of the process have created a nihilistic climate among scientists and the public. The identification of single gene alterations that substantially extend life span in nematodes and flies however, have begun to reinvigorate the field. Drosophila's long history of contributions to aging research, rich storehouse of genetic information, and powerful molecular techniques make it an excellent system for studying the molecular mechanisms underlying the process of aging. In recent years, Drosophila has been used to test current theories on aging and explore new directions of potential importance to the biology of aging. One such example is the surprising finding that, as opposed to the commonly held assumption that adult life is a period of random passive decline in which all things are thought to fall apart, the molecular life of the adult fly appears to be a state of dynamic well-regulated change. In the fly, the level of expression of many different genes changes in an invariant, often age-dependent, manner. These as well as other molecular genetic studies and demographic analyses using the fly have begun to challenge widely held ideas about aging providing evidence that aging may be a much more dynamic and malleable process than anticipated. With the enormous success that Drosophila molecular genetics has demonstrated in helping understand complex biological phenomena such as development there is much optimism that similar approaches can be adapted to assist in understanding the process of aging. PMID- 12539240 TI - Fibrillar collagen: the key to vertebrate evolution? A tale of molecular incest. AB - Fibril-forming (fibrillar) collagens are extracellular matrix proteins conserved in all multicellular animals. Vertebrate members of the fibrillar collagen family are essential for the formation of bone and teeth, tissues that characterise vertebrates. The potential role played by fibrillar collagens in vertebrate evolution has not been considered previously largely because the family has been around since the sponge and it was unclear precisely how and when those particular members now found in vertebrates first arose. We present evidence that the classical vertebrate fibrillar collagens share a single common ancestor that arose at the very dawn of the vertebrate world and prior to the associated genome duplication events. Furthermore, we present a model, 'molecular incest', that not only accounts for the characteristics of the modern day vertebrate fibrillar collagen family but demonstrates the specific effects genome or gene duplications may have on the evolution of multimeric proteins in general. PMID- 12539241 TI - The CRP/MLP/TLP family of LIM domain proteins: acting by connecting. AB - In vertebrates, members of the cysteine-rich protein (CRP) family are characterized by the presence of two LIM domains linked to short glycine-rich repeats. These proteins mediate protein-protein interactions and are of fundamental importance for cell differentiation, cytoskeletal remodeling, and transcriptional regulation. To date, a vast amount of information about vertebrate CRPs has become available, including their biological functions, interacting partners, and three-dimensional structures. Compatible with a molecular adapter role, structural data reveal that the LIM domains within these proteins represent completely independent folded units bridged by flexible linker regions. The physiological roles for individual CRPs was determined by targeted gene disruption analysis and by identification of common and specific binding partners by means of yeast and mammalian two-hybrid screens. Several CRP-like LIM domain proteins with close structural and sequence similarity were identified in arthropods, protozoas and plants, supporting the notion that this subset of LIM domain proteins has been highly conserved over the span of evolution thereby emphasizing the importance of their function. PMID- 12539242 TI - Levels of polymorphism on the sex-limited chromosome: a clue to Y from W? AB - Nucleotide diversity of the human Y chromosome is much lower than that in the rest of the genome. A new hypothesis postulates that this invariance may result from mutations in maternally inherited mitochondrial DNA (mtDNA), leading to impaired reproduction among males and lowered male effective population size. If correct, we should expect to see low levels of polymorphism in the male-specific Y chromosome of many organisms but not necessarily in the female-specific W chromosome in organisms with female heterogamety. However, recent observations from birds suggest that the avian W chromosome is very low in nucleotide diversity. This indicates that mtDNA mutations cannot broadly explain the evolution of the sex-limited chromosome. Other work has suggested that sexual selection at loci involved in sex determination or secondary sexual characteristics might reduce levels of genetic variability on Y through hitch hiking effects. Although the W chromosome does not seen to play a dominant role for sex determination in birds, it cannot be excluded that selective sweeps arising from natural or sexual selection contribute to the low levels of genetic variability seen on this chromosome. PMID- 12539243 TI - When parsimony backfires: neglecting DNA repair may doom neurons in Alzheimer's disease. AB - Taking advantage of the fact that they need not replicate their DNA, terminally differentiated neurons only repair their expressed genes and largely dispense with the burden of removing damage from most of their genome. However, they may pay a heavy price for this laxity if unforeseen circumstances, such as a pathological condition like Alzheimer's disease, cause them to re-enter the cell cycle. The lifetime accumulation of unrepaired lesions in the silent genes of neurons is likely to be significant and may result in aborting the mitotic process and triggering cell death if the cells attempt to express these dormant genes and resume DNA replication. PMID- 12539244 TI - Proline-directed phosphorylation and isomerization in mitotic regulation and in Alzheimer's Disease. AB - The reversible phosphorylation of proteins on serine/threonine residues preceding proline (Ser/Thr-Pro) is a major regulatory mechanism for the control of a series of cell cycle events. Although phosphorylation is thought to regulate protein function by inducing conformational changes, little is known about most of these conformational changes and their significance. Recent studies indicate that the conformation and function of a subset of these phosphorylated proteins are controlled by the prolyl isomerase Pin1 through isomerization of specific phosphorylated Ser/Thr-Pro bonds. Furthermore, compelling evidence supports the idea that proline-directed phosphorylation and subsequent isomerization play a critical role not only in cell cycle control, but also in the development of Alzheimer's disease, where postmitotic neurons display various cell cycle markers, especially mitotic events, prior to degeneration. PMID- 12539245 TI - A phylogenetic hypothesis for the origin of hiccough. AB - The occurrence of hiccoughs (hiccups) is very widespread and yet their neuronal origin and physiological significance are still unresolved. Several hypotheses have been proposed. Here we consider a phylogenetic perspective, starting from the concept that the ventilatory central pattern generator of lower vertebrates provides the base upon which central pattern generators of higher vertebrates develop. Hiccoughs are characterized by glottal closure during inspiration and by early development in relation to lung ventilation. They are inhibited when the concentration of inhaled CO(2) is increased and they can be abolished by the drug baclofen (an agonist of the GABA(B) receptor). These properties are shared by ventilatory motor patterns of lower vertebrates, leading to the hypothesis that hiccough is the expression of archaic motor patterns and particularly the motor pattern of gill ventilation in bimodal breathers such as most frogs. A circuit that can generate hiccoughs may persist in mammals because it has permitted the development of pattern generators for other useful functions of the pharynx and chest wall muscles, such as suckling or eupneic breathing. PMID- 12539246 TI - Correlation between prostate cancer grade and vascularity on color doppler imaging: preliminary findings. AB - PURPOSE: We examined the relationship between the amount of prostate cancer associated vascularity as seen on color Doppler imaging and the tumor grade. METHODS: Transrectal color Doppler imaging of the prostate was performed in 54 patients with prostate cancer. Color flow signal/total pixel ratios (SPRs) of selected images were calculated using the ratio of the number of pixels showing color Doppler signals to the total number of pixels within the lesion. All the patients underwent prostate biopsy guided by transrectal sonography. Gleason scores were determined from the biopsy specimens. RESULTS: Color Doppler signals were demonstrated in the lesions of 91% (49/54) of the patients. The mean SPRs of prostate cancers 3 cm or smaller and larger than 3 cm were 0.15 +/- 0.07 and 0.11 +/- 0.04 (+/- standard deviation), respectively (p < 0.05). The mean SPRs of well (Gleason score of 2-4), moderately (Gleason 5-7), and poorly (Gleason 8-10) differentiated prostate cancers were 0.08 +/- 0.03, 0.12 +/- 0.06, and 0.17 +/- 0.11, respectively (r = 0.45; p < 0.01). CONCLUSIONS: The refinement of color Doppler equipment has improved the detection of color Doppler signals associated with prostate cancer. Our study shows a correlation between prostate cancer associated vascularity as shown on color Doppler imaging and the tumor grade. PMID- 12539247 TI - Chest sonography versus lateral decubitus radiography in the diagnosis of small pleural effusions. AB - PURPOSE: The aim of this prospective study was to assess the value of chest sonography in the radiologic diagnosis of small pleural effusions (relative to expiratory lateral decubitus radiography) and to suggest gray-scale sonographic criteria for detecting the presence of small pleural effusions. METHODS: Patients referred for abdominal or chest sonographic evaluation for various reasons were also examined for sonographic features of pleural effusion from May 1, 1997, until January 31, 2000. Patients who had evidence of small pleural effusions were included. Patients with no such evidence served as a control group. Subsequently, all patients underwent erect posteroanterior and expiratory lateral decubitus chest radiography. RESULTS: On chest sonography, 52 patients were found to have small pleural effusions. The control group consisted of 17 patients. The mean thickness of the pleural effusion was 9.2 mm on sonography and 7.6 mm on expiratory lateral decubitus radiography (p < 0.01). Compared with radiologic examination, chest sonography had a positive predictive value of 92% in the diagnosis of small pleural effusions in our study population. CONCLUSIONS: Chest sonography showed a high degree of accuracy relative to that of lateral decubitus chest radiography in the diagnosis of small pleural effusions, which appeared as thin (usually 15 mm thick or less) anechoic areas that changed shape with the phases of respiration. PMID- 12539248 TI - Resistance index in fetal interlobar renal artery with renal pelvic dilatation up to 10 mm. AB - PURPOSE: The purpose of this study was to compare the resistance indices (RIs) in the fetal interlobar renal arteries (IRAs) of third-trimester fetuses with or without pelvicaliceal dilatation of up to 10 mm and to compare them with those of the full-term healthy infants. METHODS: Women with uncomplicated, low-risk, singleton third-trimester pregnancies were examined sonographically. The RIs in the IRAs were measured in the fetuses, who were stratified into 3 groups according to the anteroposterior diameter of the renal pelvic dilatation: group I, no dilatation; group II, 1-5-mm dilatation, and group III, 6-10-mm dilatation. RESULTS: In total, 178 women were examined. We could study both kidneys in 139 of the fetuses; in the other 39, only 1 kidney could be imaged perfectly. This yielded a total of 317 kidneys. Group I fetuses included 172 (54%); group II, 98 (31%); and group III, 47 (15%) of the kidneys. The mean (+/- standard deviation) RIs in the IRAs were 0.81 +/- 0.09, 0.80 +/- 0.07, and 0.80 +/- 0.06 in the 3 groups, respectively, with no statistically significant difference between the groups (p = 0.72). There was also no statistically significant difference between the RIs recorded in the right and left kidneys. The mean RI in the IRAs of the 34 infants who were available for follow-up 6-12 weeks after delivery was 0.73 +/- 0.07, which was significantly less than that recorded in the third-trimester fetuses (p = 0.005). CONCLUSIONS: The RI in the fetal IRA does not differ in fetuses with and without renal pelvic dilatation of up to 10 mm. Thus, an increase in the RI or an RI that significantly differs between the right and left kidneys should be investigated further for possible renal pathology. PMID- 12539249 TI - Sonographic measurements of normal gallbladder sizes in children. AB - PURPOSE: Our goal was to establish the range of sonographic measurements of normal gallbladders in children. METHODS: Six hundred ten children aged 0-16 years (male:female ratio, 1.5:1) with normal clinical and laboratory findings were included in this study. The sonographic parameters were the length, width, and calculated volume of the gallbladder, and the clinical parameters were the age, height, weight, and body surface area of the children. Statistical significance was determined through correlation and regression analyses. RESULTS: The length of the gallbladder showed significant positive correlations with age (r = 0.65), height (r = 0.67), weight (r = 0.63), and body surface area (r = 0.65; p < 0.01). The calculated volume of the gallbladder also showed moderate correlations with age (r = 0.53), height (r = 0.55), weight (r = 0.61), and body surface area (r = 0.57; p < 0.01). The gallbladder width showed modest but significant correlations with age (r = 0.48), height (r = 0.53), weight (r = 0.53), and body surface area (r = 0.55; p < 0.01). The highest correlation coefficients were found between the gallbladder length and subject age (r = 0.65; p < 0.01) and between the gallbladder length and subject height (r = 0.67; p < 0.01). For all correlations, statistical significance remained after regression analysis (p < 0.01). CONCLUSIONS: Values for the size of the normal pediatric gallbladder are defined and will be helpful in the diagnosis of gallbladder abnormalities. PMID- 12539250 TI - Sonographic findings in patients with anterior knee pain. PMID- 12539251 TI - Caput succedaneum mimicking a cephalocele. AB - Caput succedaneum is relatively common at birth but infrequently diagnosed in utero. It has a benign prognosis, but it is very important not to misdiagnose it as a cephalocele, which carries a guarded prognosis. We present the case of a patient who experienced preterm labor and premature rupture of the membranes at 28 weeks' menstrual age. Our initial diagnosis was cephalocele, but after the male infant was delivered by cesarean section, examination revealed no bone defect, and our final diagnosis was caput succedaneum. We also discuss the sonographic findings and diagnostic differences between caput succedaneum, cephalocele, and other fetal head masses. PMID- 12539252 TI - Imaging findings in Mondor's disease. AB - Mondor's disease, or acute thrombophlebitis of superficial veins of the breast, occurs rarely, and little information is available in the medical literature on imaging findings in this disease. We present the case of a 59-year-old woman who underwent evaluation of a palpable, painful cord in her left breast associated with discoloration of the overlying skin. Mammography revealed a 10-cm-long linear density in her left breast. Gray-scale sonography showed a noncompressible, hypoechoic, undulating tubular structure in the subcutaneous fat. Color Doppler imaging revealed no flow signal in this structure, although we observed arterial flow signals adjacent to it. We diagnosed Mondor's disease and treated the patient symptomatically; 6-week follow-up examinations demonstrated resolution of the venous thrombosis. The use of sonography, particularly color Doppler imaging, can aid in diagnosing Mondor's disease and in monitoring its resolution. PMID- 12539253 TI - Sonographic detection of a malpositioned feeding tube causing esophageal perforation in a neonate. AB - We report a case of esophageal perforation caused by a malpositioned feeding tube in a neonate of extremely low birth weight, 632 g. The infant had respiratory distress, which increased rapidly when he was 6 days old. Radiography revealed right-sided hydrothorax that had not been evident a day earlier but no sign of a perforated esophagus. We performed sonography, which revealed fluid in the right pleural cavity and extra-esophageal placement of the feeding tube. Analysis of a fluid specimen obtained on thoracocentesis indicated that the fluid was feeding formula. The feeding tube's misplacement was confirmed sonographically by injecting a small amount of sterile distilled water into the tube and visualizing its entry into the pleural cavity. The feeding tube was removed, and antimicrobial agents were administered. When the infant was 15 days old, feeding resumed through another tube, the placement of which was verified radiographically. The infant was discharged when he was 118 days old with no severe complications, although he had mild chronic lung disease. Because radiography did not reveal the tube's misplacement in this case, we believe that the use of sonography can contribute to an early diagnosis of esophageal perforation in such cases. PMID- 12539254 TI - Gray-scale and color Doppler findings in bilateral internal jugular vein thrombosis caused by anaplastic carcinoma of the thyroid. AB - Bilateral internal jugular vein thrombosis is a rare complication of thyroid cancer. The most common manifestation of this condition is superior vena cava syndrome. We report the sonographic findings in a case of bilateral internal jugular vein thrombosis with mild symptoms. There was evidence of direct infiltration of anaplastic thyroid carcinoma into the left internal jugular vein. Sonographic and CT examinations also demonstrated multiple dilated collateral veins. PMID- 12539255 TI - Infantile scrotal pyocele simulating missed testicular torsion on sonography. AB - We present the case of a neonate with idiopathic infantile pyocele. Scrotal sonography revealed a complex fluid collection within the left scrotal sac containing septations and a fluid-fluid level. The dependent region of the collection had moderate echogenicity, the slightly hypoechoic testis was not well defined, and the scrotal wall was thickened. Color Doppler sonography revealed mild hypervascularity in the thickened scrotal wall but no vascularity inside the testis. The sonographic findings suggested missed testicular torsion, but surgery revealed a pyocele, for which no source was identified. Radiologists should be aware that idiopathic infantile pyocele can mimic the Doppler sonographic findings in missed testicular torsion. PMID- 12539257 TI - X-ray studies on ligands. AB - Advances in x-ray crystallographic data collection, structure solution, and refinement/validation have reduced the time required and expanded the range of samples amenable to x-ray crystallographic studies. Consequently, we can now collect complete atomic resolution data sets on physically smaller crystals and solve larger problems by direct methods beyond what could have been accomplished even five years ago. Applying these improved methods to the study of opioid ligands has enhanced our knowledge of the opioid pharmacophore. Despite considerable progress, it is still difficult to define the pharmacophoric parameters required for highly selective and potent opioid peptides. In part this is due to the conformational flexibility remaining even in conformationally constrained peptides. PMID- 12539258 TI - Regulation of the K channels by cytoplasmic domains. AB - Regulation of intracellular potassium levels is one of the basic functions of all cells, controlling cellular osmolarity and transmitting information. In higher organisms, elaborate control of transmembrane potassium flux has evolved to endow nervous systems with the remarkable ability to transmit electrical signals between cells. Multiple genes, gene splicing, mRNA editing, and selective tetrameric assembly of K channel genes provide the basis for creating distinct electrophysiological properties at varying developmental and cellular stages. This assembly mechanism, primarily governed by the T1 domain, is under the control of intracellular signals. Atomic structures of the isolated T1 domains of Shaker and Shaw subfamilies provided us with valuable structural insights into understanding both channel assembly and functional regulation of the entire channel molecule through conformational changes. PMID- 12539259 TI - Crystallization of membrane proteins from media composed of connected-bilayer gels. AB - The use of hydrated-lipid gels in which the bilayer is an infinitely periodic (or at least continuous), three-dimensional structure offers a relatively new approach for the crystallization of membrane proteins. While excellent crystals of the Halobacterial rhodopsins have been obtained with such media, success remains poor in extending their use to other membrane proteins. Experience with crystallization of bacteriorhodopsin has led us to recognize a number of improvements that can be made in the use of such hydrated-gel media, which may now prove to be of general value for the crystallization of other membrane proteins. PMID- 12539260 TI - Structural models for dimerization of G-protein coupled receptors: the opioid receptor homodimers. AB - Among the most exciting functional features of G-protein coupled receptors (GPCRs) that are coming into focus lately are those relating to the role and structural characteristics of their oligomerization (mostly homo- and heterodimers). The structural underpinnings of these novel functional insights are still not clear, as current experimental techniques have not yet succeeded in identifying the dimerization interfaces between GPCR monomers. Two computational approaches have recently been designed in our lab to provide reasonable three dimensional (3D) molecular models of the transmembrane (TM) regions of GPCR dimers based on a combination of the structural information of receptor monomers and analyses of correlated mutations in receptor families. The modeling of GPCR heterodimers has been described recently. We present here a related approach for modeling of GPCR homodimers that identifies the interfaces in the most likely configurations of the complexes. The approach is illustrated for the three cloned opioid receptor subtypes (OPRD, OPRM, and OPRK). PMID- 12539261 TI - Studies of the receptor-bound conformation of alphaIIbbeta3 antagonists by 15N edited NMR spectroscopy. AB - We report the results of NMR studies and computer simulations of potent antagonists reflective of the alpha(IIb)beta(3) receptor-bound conformations. The peptides c[Mpa-(15)N-Arg(1)-(15)N-Gly(2)-(15)N-Asp(3)-(15)N-Phe(4)-(15)N-Arg(5) Cys]-NH(2) (Phe-Arg analog) (Mpa: 3-mercaptopropionic acid) and c[Mpa-(15)N Arg(1)-(15)N-Gly(2)-(15)N-Asp(3)-(15)N-Asp(4)-(15)N-Val(5)-Cys]-NH(2) (Asp-Val analog) were subjected to (15)N-edited NMR experiments to study the conformations of these peptides in the absence and in the presence of alpha(IIb)beta(3) receptor. The NMR studies of the Phe-Arg analog, a selective alpha(IIb)beta(3) antagonist, resulted in distinctly different experimental data in the presence and absence of the receptor. The computer simulations for this peptide resulted in one large family of structures consistent with the experimental data. This conformation suggests a type I beta-turn spanning residues Arg(1) and Gly(2) when bound to the receptor and we were able to establish a model for the three dimensional arrangement of the pharmacophores. The studies on the Asp-Val analog, an alpha(v)beta(3) antagonist that binds to the alpha(IIb)beta(3) with moderate affinity, resulted in conformations that are not as well defined as those for the Phe-Arg analog but are consistent with the model established for this analog. These results are important for the design of novel alpha(IIb)beta(3) antagonists. PMID- 12539262 TI - Extracellular domains of the neurokinin-1 receptor: structural characterization and interactions with substance P. AB - The technical difficulties associated with the structure determination of membrane proteins have limited the structural information available for the ligand binding to G-protein coupled receptors (GPCRs). Here, we describe a reductionist approach to GPCR structure determination in which the extracellular domains of the receptor are examined by high-resolution NMR in the presence of a membrane mimetic. The resulting structural features are then incorporated into a molecular model of the receptor, utilizing the x-ray structure of rhodopsin to generate the topological orientation of the transmembrane helices. The results of our study of the neurokinin-1 receptor (NK-1R) and its interactions with substance P (SP) are detailed here. The structure of the N-terminus, NK-1R(1-39), and of the third extracellular loop, NK-1R(264-290), in the presence of dodecylphosphocholine micelles is described. Our findings provide a structural basis for the interpretation of the results from other methods including mutagenesis, fluorescence, and photoaffinity labeling experiments, resulting in an experimentally based, high-resolution model of SP binding to NK-1R. PMID- 12539263 TI - Polar mutations in membrane proteins as a biophysical basis for disease. AB - Transmembrane (TM) alpha-helices are surrounded by the hydrocarbon chains of the lipid bilayer. The low dielectric constant of this environment makes it extremely unfavorable for a residue with a polar side chain to exist in a non-H-bonded state. Therefore, in combination with a wild-type polar residue partner, a polar TM mutant could generate, in some cases, a non-native H-bond that could impair native protein structure/function-and possibly lead to a disease state. We have examined protein mutation databases and have found many examples of TM-based apolar to polar mutations that are, in fact, a cause of human disease. Here we review the various molecular defects that such mutations can produce, including impeding protein dynamics by side-chain-side-chain interhelical H-bond cross links; alteration of helical packing through steric hindrance; and disruption of a protein active site. We further note that the reverse case--membrane-embedded polar to apolar mutations--can similarly cause human disease, implying that native interhelical H-bonds can also play pivotal roles in stabilizing native TM domains. As a specific example, we show that the Gly to Arg mutation occurs statistically more frequently in TM domains as compared to its occurrence in soluble domains, suggesting that TM-based G-to-R mutations have a high "phenotypic propensity" for disease. A more complete understanding of how mutations involving polar residues in TM domains of proteins translate into compromised function may aid in the development of novel therapeutics. PMID- 12539265 TI - [Improving and developing neuropharmacological research methods] PMID- 12539266 TI - [Novel quantitative method for evaluating oxygen/glucose deprivation-induced injury of hippocampal slices] AB - OBJECTIVE: To establish a simple, sensitive in vitro method to evaluate oxygen/glucose deprivation (OGD)-induced injury of brain hippocampal slices in rats. METHODS: Rat hippocampal slices were incubated in 2% 2, 3, 5 triphenyltetrazolium chloride (TTC) solution after oxygen/glucose deprivation. They were then soaked in a measured volume of ethanol and dimethylsulfoxide (50:50) to extract the TTC formazan Product which was then measured by spectrophotometry. OGD induced LDH release was simultaneously measured. RESULTS: Progressive prolongation of OGD induced hippocampal injury resulted in decreased formazan coloration as determined by spectrophotometry. There was a parallel increase in LDH release, thus a negative correlation in these two products was noted. (r=-0.933,P <0.01). The injury was attenuated in the brain slices pre treated with nimodipine, dexamethasone, and ketamine, but not ONO-1078. CONCLUSION: Solvent extraction and spectrophotometric quantification of formazan represents an objective measurement of OGD-induced injury of rat hippocampal slices. PMID- 12539264 TI - Glycine receptors: lessons on topology and structural effects of the lipid bilayer. AB - The members of the superfamily of nicotinicoid receptors, sometimes referred to as the ligand-gated ion channel superfamily (LGICS), are essential mediators in the propagation of electrical signals between cells at neuronal and neuromuscular synapses. Given the significant sequence and proposed topological similarities between family members, the structural architecture of any one of these neuroreceptors is believed to be archetypic for the family of ligand-gated channels. We have focused our biophysical studies on the glycine receptor (GlyR) since homomeric expression of just the alpha1 chain of the receptor is sufficient to reconstitute native-like activity when expressed in heterologous cells, and we have successfully overexpressed and purified relatively large quantities of this receptor. Our CD data suggests that the historical four transmembrane helix topology model for nicotinicoid receptors may be erroneous. Proteolytic studies as well as chemical modification studies coupled with mass spectroscopy (MS) have provided additional evidence that this model may be inappropriate. While we suggest a novel topological model for the superfamily of nicotinicoid receptors, the absence of high resolution data for the transmembrane regions of these ion channels precludes further refinement of this model. In addition, we observe structural changes in the recombinant alpha1 GlyR as a function of bilayer composition, suggesting that these receptors may be dynamically modulated by cellular control over the properties of the plasma membrane. PMID- 12539267 TI - [Light transmission measurement of focal ischemic cerebral infarction in mice] AB - OBJECTIVE: To evaluate the feasibility of light transmission to measure focal cerebral ischemia in mice. METHODS: Persistent focal cerebral ischemia was induced by middle cerebral artey occlusion (MCAO) in mice. The brain were removed 24 h after MCAO and coronally dissected into 1 mm sections. Using a stereomicroscope, the brain section was illuminated with a halogen lamp and computerized images were stored. Next the brain sections were stained for 30 minutes with 0.5% TTC (2, 3, 5-triphenylterzolim chloride) at 37 degrees C. Using an image analyzer (AnalyPower 1.0), the infarct volumes obtained by light transmittance and TTC staining were calculated. Integrated gray scales of sections of both hemispheres were calculated by Photoshop 5.0. RESULTS: A close correlation existed between cerebral infarct volume measured by light transmission and TTC staining (r=0.81). The mean gray scales measured by both techniques of the ischemic hemispheres as well as those of the cortex, subcortex and hippocampus were siginificantly higher than those of non-ischemic hemispheres and of control mouse hemispheres (P <0.001). Further there were no significant difference between the two hemispheres of control mice and between hemispheres of control mice and non-ischemic hemispheres of the MCAO mice. CONCLUSION: Light transmission can be used for qualitative analysis of focal cerebral ischemia. PMID- 12539268 TI - [Optical recording method for evaluation of neuronal damage in rat hippocampal slices during ischemia and reperfusion] AB - OBJECTIVE: To develop a novel technique of optical recording and its validation for assessment of the neuroprotective effect of nimodipine, a L-type calcium channel blocker. METHODS: In vitro ischemia was induced by oxygen/glucose deprivation (OGD), the light transmittance (LT) of rat hippocampal slices undergoing OGD and reperfusion was quantitated using a simple apparatus relying on basic principles of light transmittance and a computerised image analysis system. RESULTS: OGD was associated with increased LT in the stratum radiatum of CA1 area and the dentate gyrus in hippocampal slices. Peak LT occurred (7.59 +/ 1.42) min after OGD, followed by a marked decrease in LT (n=15 slices). Nimodipine administration (0.5 &mgr;mol/L, n=10 slices, 5 &mgr;mol/L, n=9 slices) appeared to protect the tissue from OGD damage by inhibiting elevation of LT, However, 50 &mgr;mol/L nimodipine resulted in increased LT (25.83 +/-6.32). min after administration (n=11 slices). CONCLUSION: LT signal measurement is a non invasive, reliable method for determination of neuronal damage in ischemic rat brain slices Nimodipine is demonstrated opposite neuroprotective effects depending on its dose. PMID- 12539269 TI - [Effects of TAK-147, a novel acetylcholinesterase inhibitor, on spatial memory deficit as evaluated by Morris water maze of rats] AB - OBJECTIVE: To evalute the effects of TAK-147, a novel acetylcholinesterase inhibitor on rat spatial memory deficit using the Morris water maze. METHODS: Morris water maze was used to measure spatial memory in rats, and open field test was used to analysis locomotor activity. RESULTS: Scopolamine (0.4mg/kg,IP) significantly increased the latency period in memory acquisition. Intraperitoneal TAK-147 injection ameliorated scopolamine-induced deficit in a dose-related manner. A significant effect was obtained at doses of 0.3 and 1.0 mg/kg. Both TAK 147 (0.3 and 1.0 mg/kg) and tacrine (3 and 5 mg/kg) significantly reversed scopolamine (1.5 mg/kg) increased latency in memory retrieval. However, TAK-147 had a more potent effect than tacrine. In the locomotor test, TAK-147 created no appreciable change, compared with scopolamine or saline. CONCLUSION: A novel acetycholinesterase inhibitor, TAK-147 ameliorates the scopolamine induced impaired spatial memory in rats. PMID- 12539270 TI - [Sedative and anticonvulsant effect of jujuboside A] AB - OBJECTIVE: To study the sedative effects of Jujuboside A (JuA) on the Central Nervous System of mice. METHODS: Using a novel jiggle-cage test, we compared the sedative effect of JuA with that of Diazepam (DZP) both with a single and cumulative dose of JuA. We also assessed the anticonvulsant effect of JuA on pentylenetetrazol (PTZ)-induced seizures in mice. RESULTS: JuA significantly decreased total activity intensity and increased the quiet state time of mice. The sedative effects of JuA were more stable and more lasting than that of DZP. However, JuA failed to resist and delay the induced seizure activity in mice. CONCLUSION: Though JuA has sedative effects on mice CNS, it has no anticonvulsant effect on PTZ-induced seizures. PMID- 12539271 TI - [A case-control study of risk factors for childhood cerebal palsy] AB - OBJECTIVE: To identify possible risk factors for cerebral palsy (CP) in children. METHODS: A Population-based survey was conducted (including 92 CP cases) in 66 townships of 15 cities of Zhejiang Province from October to November, 1998. 184 of matched controls were selected for comparison. RESULTS: Factors identified which were statistically significant for risk of subsequent childhood Cerebral Palsy included some neonatal diseases, some maternal diseases, low birth weight (<2500 g), maternal irregular menstruation, toxic, substances during pregnancy, malnutrition during pregnancy,and paternal age. CONCLUSION: Several risk factors for Cerebral Palsy were identified. Their prevention may result in redduction of the incidence of Cerebral Palsy. PMID- 12539272 TI - [C-Fos expression in the hippocampus of rats with pentylenetetrazol-induced epilepsy] AB - OBJECTIVE: To investigate the expression of c-FOS oncogene in rats hippocampus with pentylenetetrazol (PTZ)-induced generalized seizure. To also investigate the anticonvulsant effect of calcium antagonists. METHODS: 52 rats was divided into 6 groups, 12 received 0.9% saline as control, 21 received PTZ, and 19 received both Nimodipine and PTZ. PTZ was injected intraperitoneally and the time to onset of seizure occurrence was recorded. The seizure events were scored using the Racine scale (1972). The c-FOS protein expression was observed at 2 h or 4 h using peroxidase-labelled streptavidin biotin (LSAB) staining techniques. RESULTS: Control antimals had no seizure activity. There was a significant increase in the time to onset of seizure activity in the Nimodipine-treated group, compared with that of the PTZ group (P<0.01). Seizure activities was more severe in the PTZ group compared with rats who received both PTZ and Nimodipine [P<0.01]. There was low level c-FOS protein expression in hippocampal pyramidal neurons and granule cells of the dentate gyrus.C-FOS expression was upregulated at the 2nd and 4th hour post-PTZ injection. Nimodipine could reduce c-FOS protein expression induced by the PTZ-induced generalized seizure at 2nd hour post-PTZ injection (P<0.01). There was no significant difference in c-FOS expression between the PTZ group and the Nimodipine-treared group by 4th hour (P> 0.05). CONCLUSION: PTZ can induce generalized seizure in rat. There is post-ictal upregulation of c-FOS protein expression in the hippocampal pyramidal neurons and dentate gyrus granule cells. Thus the hippocampus pathways are likely involved in the occurrence of PTZ induced epilepsy. Nimodipine may attenuate PTZ-induced seizure activity. PMID- 12539273 TI - [Effects of intrasplenic transplantation of IL-18 gene-modified fetal hepatocytes on mouse immune function] AB - OBJECTIVE: To investigate the effects of IL-18 gene-modified fetal hepatocytes (AdmIL-18/MNL.CL2) intrasplenic transplantation on mouse immune function. METHODS: Forty mice were evenly divided into 4 groups of 10. Each group received an intrasplenic transplantation one of the following: AdmIL-18/BNL.CL2, Ad LacZ/BNL.CL2 (virus control), BNL.CL2 (cell control) and PBS (blank control). After two weeks, the mice were sacrificed. Serum cytokine levels, Mpsi and splenic cell culture supernatant and liver tissue extracts supernatants were measured using ELISA. Hepatic cytokines mRNA expression were determined by RT PCR. THe cytotoxicity of peritoneal Mpsi and NK activity of spienocytes were detected by LDH release assay. The proliferation of splenic lymphocytes was determined by MTT assay. RESULTS: The IL-18, IL-2,IFN-gamma, TNF-alpha levels of serum, Mpsi and splenocyte culture supernatant, liver tissue extracts supernatants in mice transplanted with AdmIL-18/BNL.CL2 were higher and the IL-4, IL-10 levels were lower compared to their levels in other 3 groups. The highest IL-18, IL-2, IFN-gamma, TNF-alpha and the lowest IL-4, IL-10 mRNA expressions in the liver were observed in mice transplanted with AdmIL-18/BNL.CL2. The mice transplanted with AdmIL-18/BNL.Cl2 showed significantly increases cytotoxicity of Mpsi, NK activity and splenic cell proliferation compared with the other 3 groups. CONCLUSION: AdmIL-18 can be effectively transfected into mice fetal heptocytes which subsequently IL-18. Intransplenic transplantation of IL-18 gene modified fetal hepatocytes may augment mouse immune function and provide an useful basis for targeted gene therapy of liver disease. PMID- 12539275 TI - [Apoptosis peripheral of T lymphocytes in patients with acute exacerbration of chronic bronchitis] AB - OBJECTIVE: To compare peripheral T lymphocyte apoptosis of chronic bronchitis during an acute episode with that during a stable period. METHODS: Seventeen Chronic Btonchitis patients who presented to the ER with an acute episode were compared with 10 stable Chronic Bronchitis patients seen in the out-patient clinic. Peripheral venous blood was obtained from each patient and T lymphocytes were separated by fluorescent monoclonal antibodies. CD3, CD95 and CD25 were measured by flow cytometry. RESULTS: In the acute episode patients, the total quantity of T lymphocytes with CD3 was (42.63 +/-16.37) and with CD95 (14.15 +/ 11.06). In the stable group T lymphocyte CD3 was (64.22 +/-23.90) and CD95 (34.88 +/-8.92) (t=-4.238, P <0.001; t=-8.796, P <0.001). The total quantity of Lymphocyte CD25 did not differ significantly between the two groups (P > 0.05). Total quantity of active T lymphocytes with CD3/CD25 was lower in the acute episode group (6.26 +/-3.94) compared to the stable group (10.52 +/-3143) (t= 1.068, P <0.05). CONCLUSION: T lymphocyte apoptosis increased in Chronic Bronchitis patients who presented with an acute episode. They also experienced a significant disturbance in immune function. PMID- 12539276 TI - [Cyclic nucleotides phosphodiesterase activity in a rat lung model of asthma] AB - OBJECTIVE: To assess the change of cGMP-PDE and cAMP-PDE activity in a rat lung model of asthma. METHODS: cGMP-PDE and cAMP-PDE activity were determined by HPLC. RESULTS: Both cAMP-PDE activity and cGMP-PDE activity in the lung tissue of antigen-challenged rats were higher than that from the normal rats (P <0.05). CONCLUSION: In the rat lung model of asthma, cyclic nucleotides phosphodiesterase activity was elevated. This may be significant in the pathogenesis of asthma. PMID- 12539277 TI - [Anti-tussive and expectorant effects of Liangyuan Pipagao,a Chinese medicine] AB - OBJECTIVE: To evaluate the pharmacological effects of Liangyuan Pipagao on cough reflex and ciliary action. Liangyuan Pipagao is a compound preparation of traditional Chinese medicine. METHODS: Cough was induced by aerosol citric acid in guinea pigs and aerosol capsacin in mice. Excretion function of the airway in mice was determined by phenol red method. Ciliary movement function of frog esophagus was examined by a migration method of charcoal granules. RESULTS: Liangyuan Pipagao inhibited both the citric acid-induced cough in guinea pigs and capsacin-induced cough in mice. ID(50)value 2.64 g/kg (95%Cl1.12 approximately 6.19) and 11.40 g/kg (95%Cl5.76 approximately 22.58) respectively. Further, Liangyuan Pipagao increased phenol red excretion in mice airways and stimulated ciliary action of frog esophagusin a dose-dependent fashion. ED(50) value 7.70 g/kg (95%Cl 4.62 approximately 12.83) and EC(25) value 1.07 X 10(-4) (95% Cl 0.394 approximately 2.92x10(-4)) respectively. CONCLUSION: Liangyuan Pipagao a traditional Chinese medicine may have anti-tussive as well as expectorant actions. PMID- 12539278 TI - [Restoration of peristalsis in postcholecystectomy patient under self-controlled analgesia by extradural anesthesia] PMID- 12539279 TI - [Clincal study of transesophageal cardioversion] PMID- 12539280 TI - [Recent development of study on the potential role of lycopene for human health] PMID- 12539281 TI - [Cytocompatibility of three substituted materials of human beings' hard tissue]. AB - OBJECTIVE: To evaluate the osteocompatibility of three substituted materials of human beings' hard tissue (hydroxyapatite, bioglass ceramic and Ti-6Al-4V) and a new experimental method for evaluating the osteocompatibility of replaced materials of hard tissues in vitro. METHODS: The isolated osteoblasts from parietal and frontal bones of Sparague Dawley rats from 1 day to 3 days were used to estimate osteocompatibility of three substituted materials in vitro. The cells were inoculated onto specimens of HA, BGC and Ti-6Al-4V for 1 day, 3, 5 and 7 days, then the morphology and proliferating rates of cells on different materials were tested. RESULTS: Rat osteoblasts could attach, spread and proliferate on three materials as normal cells, and proliferating rates of cells on HA and BGC surface were as the same as normal, but that on Ti-6Al-4V was lower than the normal cells. CONCLUSION: The osteocompatibility of HA and BGC are better than that of Ti-6Al-4V and rat osteoblast culture system can be used to evaluate the osteocompatibility of substituted materials in vitro. PMID- 12539282 TI - [An experimental study on transplantation of frozen autogenous tooth-mandible compound]. AB - OBJECTIVE: To study the feasibility of reserving the teeth in the transplantation of frozen autogenous mandible. METHODS: Forty adult Wistar rats were used for these operations of transplantation of frozen autogenous tooth-mandible compound (ATMC) and fresh ATMC. Then the healing process after transplantation of frozen ATMC and fresh ATMC was compared by clinical, radiographical, histological and microangiographical observation. RESULTS: There were no significant differences between the healing process of frozen ATMC and that of fresh ATMC, and the fibres of regenerated periodontal membrane which maintained the dental root in normal position could be imbedded in the newly formed cementum. CONCLUSION: It is feasible to transplant the frozen ATMC for reconstructing the mandible and improving its function in clinic. PMID- 12539283 TI - [Role of apoptosis and bcl-2 gene in the temporomandibular joint development]. AB - OBJECTIVE: Apoptosis involving in embryogenesis and organ formation has been verified, but the possible roles of apoptosis and its related regulative genes have not been clarified. Temporomandibular joint dysfunction (TMD) has increased in incidence and occurred increasingly in young patients. Thus, research on the development and gene modulation is necessary. METHODS: Tdt-mediated dUTP nick end labeling (TUNEL) and mRNA in situ hybridization were used in observation of apoptosis and bcl-2 gene expression in TMJ of SD rats during prenatal and postnatal developmental period. RESULTS: Apoptosis involved in different development periods of condylar cartilage, and the cells of apoptosis mainly located in the proliferative zone and the area between the prehypertrophic zone and the late hypertrophic zone., bcl-2 was expressed in chondrocytes throughout condylar cartilage with high levels in proliferative and prehypertrophic chondrocytes and obvious low levels in the late hypertrophic chondrocytes. Very few matured chondrocytes expressed bcl-2. CONCLUSION: Apoptosis regulated by bcl 2 plays an important role in prenatal and postnatal development of TMJ. PMID- 12539284 TI - [Changes of simple epithelial cytokeratin during oral carcinogenesis]. AB - OBJECTIVE: To investigate whether the simple epithelial cytokeratin CK18 and CK19 can be used as a marker of oral precancerous lesions. METHODS: Formalin-fixed, paraffin-embedded tissue sections of normal oral mucosa, epithelial hyperplasia, mild epithelial dysplasia, moderate epithelial dysplasia, severe epithelial dysplasia and oral squamous cell carcinomas were stained with a CK18-specific antibody and CK19-specific antibody respectively by LSAB immunohistochemical method. The stained sections were observed under light microscopy. The results were described and analyzed with Rank Sum Test. RESULTS: CK18 was not detected in normal and abnormal oral tissue sections. But in normal nonkeratinized mucosa, CK19 was detected in the basal cell layer dispersively. In epithelial dysplasia, CK19 was detected in the suprabasal cell layer and the number of CK19-positive cell layers was correlated with the dysplasia degree of epithelia. Furthermore, CK19 was detected in oral squamous cell carcinoma, especially in the poor differentiated cancer cells. CONCLUSION: CK19 expression in suprabasal cell layer of oral mucosa can be used as a candidate marker for diagnosis of oral precancerous lesions and determination of the differentiation level of oral squamous cell carcinoma. PMID- 12539285 TI - [Experimental study on head and neck hemodynamics after ligation of jugular veins]. AB - OBJECTIVE: To study alteration of neck hemodynamics after ligation of jugular veins during radical neck dissection. METHODS: 4 rhesus monkeys were selected to simulate the human bilateral radical neck dissection. After operation, the jugular veins were ligated or loosen and the blood stream quantities and blood pressure of jugular veins and arteries were measured simultaneously. RESULTS: The hemodynamic index would changed after jugular vein was ligated, the more the jugular veins were ligated, the greater effects the hemodynamics index of jugular veins were occurred, but there was little effect on arteries. CONCLUSION: The external jugular veins have the effect of compensating cerebral venous outflow after removing the internal jugular veins, and the external jugular veins should be conserved when bilateral or unilateral radical neck dissection was done. PMID- 12539286 TI - [Study of relative factors which influenced the embolic level of microspheres in the hamster lingual carcinoma]. AB - OBJECTIVE: To study the relative factors which influence the embolic level of microspheres in the hamster lingual carcinoma. METHODS: Micropicture analysis instrument was used to measure the embolic level of microspheres which were poured through ascending aorta. RESULTS: The optimum pressure of embolization was 30 kPa. The embolic level approached to remote site of the lingual artery as tongue cancer developed. In early invasive cancer period, the embolic level of microspheres was at arteriole level where microspheres block "the total values" which controlled the tumor microcirculation. The optimum embolic result and less offshoot microcirculation were obtained at this time. When in later period, microspheres mainly embolized the meta-arteriole. As the microvessels dilated, the dose of microspheres must be increased to get the best embolic result. CONCLUSION: The experimental data for clinical choice of optimum pouring pressure, optimum opportunity and optimum cases. PMID- 12539287 TI - [Studies on distribution of magnetic-bearing microspheres in rabbits]. AB - OBJECTIVE: To evaluate the distribution of magnetic-bearing microspheres (MPMs) in rabbits. METHODS: The distribution of intravenously injected 99mTc-labelled MPMs in rabbits at different intervals of magnetic field applied and different magnetic field intensity was investigated by using an externally applied magnetic field and measuring the radio-activity at the rabbit head and other organs. RESULTS: MPMs have the properties of targeted distribution, as positive correlation between targetic radio-activity and times and intensity of magnetic field. CONCLUSION: MPMs may be better used in the therapy of cavernous hemangiomas. PMID- 12539288 TI - [Effects of monoclonal antibody against the major cell surface protein on salivary agglutinin mediated aggregation of Streptococcus sobrinus]. AB - OBJECTIVE: To investigate the effects of monoclonal antibody on salivary agglutinin-mediated aggregation of Streptococcus sobrinus in vitro. METHODS: Bacterial aggregation inhibition was measured under OD650 at 10 minutes intervals for 1 hour. RESULTS: It showed that the McAb ZS2/286 raised against the PAg of S. sobrinus 6715 could inhibit the aggregation by some degree, but the effect was not significant. CONCLUSION: The McAb ZS2/286 might not bind directly to the functionally important region of PAg related to aggregation. PMID- 12539289 TI - [Recombinant Salmonella typhimurium anticaries vaccine. I. Sequence of Streptococcus mutans surface protein pac gene's upstream]. AB - OBJECTIVE: To sequence the Streptococcus mutans surface protein pac gene's upstream. METHODS: The PCR autosequence kit and autosequence instrument ABI310 were used for sequence. RESULTS: The 517 base pairs sequence of the pac gene's upstream is sequenced. CONCLUSION: The sequence of pac gene's upstream will provide useful information to construct the recombinant Salmonella typhimurium anticaries vaccine. PMID- 12539290 TI - [Isolation, purification and antibacterial activities of salivary histidine-rich polypeptides]. AB - OBJECTIVE: To isolate and purify salivary histidine-rich-polypeptides (HRPs) and observe their antibacterial activities. METHODS: By using of preparative acid urea polyacrylamide gel electrophoresis (AU-PAGE), three major HRPs(HRP-1, HRP-3 and HRP-5) were purified from parotid saliva collected from healthy adults. Their antibacterial activities were tested in the ultrasensitive antibacterial assay, named radial diffusion assay. RESULTS: Each fraction of the three purified HRPs appeared as a single protein zone both in AU-PAGE and Tricine-SDS-PAGE. Their molecular weights are 3-5 kD as determined by Tricine-SDS-PAGE. All the three major HRPs showed potent bactericidal effects on Streptococcus mutans MT6R and Staphylococcus aureas ATCC25923. As to Escherichia coli ML-35P and Pseudomonas aeruginosa ATCC27853, HRP-3 and HRP-5 also demonstrated significant antibacterial activities, whereas HRP-1 did not show any inhibitory effects on the viability of them. CONCLUSION: The above results suggested that the method of preparative AU PAGE we used in this study has the benefits of high resolution, simple manipulation and perfect results of purification. HRPs are important components to prevent oral tissue from bacterial infections, therefore play a potent role in the maintenance of the oral health. PMID- 12539291 TI - [Histologic comparison of treating root surfaces with different acids]. AB - OBJECTIVE: To study the healing mechanism of different etching agents for periodontitis-affected teeth in animal models. METHODS: The animal models of defect alveolar bone were prepared, and the isolated teeth with periodontitis were scaled according to periodontal surgery in vitro. After different acid acid treatment, the dentin blocks were implanted into defect bones, then the results were examined histologically after 3, 7, 14, 28 days respectively. RESULTS: There was less inflammatory reaction in tetracycline group after operation. At the later stages, the healing in tetracycline group was characterized by osseointegration without resorption. But hydrochloric acid group and citric acid group showed different reactions. CONCLUSION: Periodontal tissue repair differs with the use of different acids. PMID- 12539292 TI - [Research on inhibition of areca nut extract on human buccal keratinocytes]. AB - OBJECTIVE: To culture human oral mucosa epithelial cells in vitro and study the mechanism of changes in the epithelia of oral submucous fibrosis (OSF). METHODS: The keratinocytes were obtained from normal buccal mucosa (NM-KC) and OSF buccal mucosa (OSF-KC) with dispase trypsine and cultured in vitro. Then the cell proliferation of keratinocytes incubated with or without areca nut extract in the presence of 10% fetal calf serum for 48 hours at 37 degrees C in 5% CO2 and air were monitored by use of thiazolyl blue (MTT) assay. RESULTS: Primary culture grew in small islands that coalesced at confluency. Ultrastructurally the cells contained a large of tonofilament and microvilli. Immunohistochemistry demonstrated uniform staining of the cells with antibodies to keratins. There was no difference in cell proliferation between OSF-KC and NM-KC, and areca nut extract inhibited keratinocyte proliferation in a concentration-dependent manner. CONCLUSION: The results suggested that utilizing dispase II can establish fibroblast-free culture for growing of human oral mucosa keratinocytes and areca nut extract is a cytotoxic agent to human buccal keratinocytes. PMID- 12539294 TI - [Application of intravenous and inhalational combined anesthesia with midazolam, fentanyl 1 and enflurane in cleft palate repair operation]. AB - OBJECTIVE: To observe the effects of intravenous and inhalational combined anesthesia with midazolam, fetanyl l and enflurane in the cleft palate repair operation. METHODS: 128 patients prepared for cleft palate repair operation were selected randomly. Anesthesia was induced with 0.2-0.3 mg/kg midazolam, 3-5 micrograms/kg fentanyl 1, and intubation was facilitated with 1-2 mg/kg succinylcholine. Anesthesia was maintained with fentanyl 1 and enflurane, and changes of hemodynamics and incident rates of hiccup, cough and vomit were observed during and after operation. RESULTS: 1. After inducing, systolic pressure (SP) was decreased by 5%-8%, diastolic pressure (DP) decreased by 8% 10%(P < 0.05), and there were no significant changes in heart rates (HR) (P > 0.05). After intubation, there were no significant changes in SP, DP and HR. 2. After induction, the incident rates of hiccup and cough were 3.1% and 7.8% respectively. After operation, the incident rate of vomit was 7.0%. 3. The volume of lost blood was 5% less than the patient's whole blood volume. CONCLUSION: The anesthetic way is easy to be controlled with few side effects, and patients can revive easily, so it is a safe, efficient and perfect way in cleft palate repair operation. PMID- 12539293 TI - [Immunohistochemical study of basement membrane proteins expression in adenoid cystic carcinoma of salivary glands]. AB - OBJECTIVE: To investigate the expression of the basement membrane proteins in adenoid cystic carcinoma (ACC) of the salivary glands and discuss the relationship between the expression of the proteins and the differentiation as well the prognosis of ACC. METHODS: 101 patients which were confirmed pathologically were collected and made a follow-up, and 30 cases of them were selected to detect the basement membrane proteins (type IV collagen, fibronectin, laminin and heparan sulfate proteoglycan) by the means of immunohistochemistry. RESULTS: The cribriform pattern of ACC had a more favorable prognosis than the solid pattern which had a lower rates of metastasis of/and recurrence. The positive locations and the degrees of the four proteins varied with the histological patterns of ACC. The cribriform pattern had the strongest positive expression and the solid pattern the weakest. In addition, the staining of fibronectin were weaker in all three patterns of ACC. CONCLUSION: The degree of the positive expression of the four proteins has a negative relation to the differentiation of the tumor cells. The lower degree of differentiation the tumor cells are, the weaker positive signals they express. Of the four substances in the basement membrane, laminin is the main material which adhere the parenchymal cell to the collagen in basement membrane of the tumor. Basement membrane proteins could be regarded as one of the basis to evaluate the prognosis of ACC. PMID- 12539295 TI - [One-time operation in infant cleft lip and palate plus alveolar process]. AB - OBJECTIVE: To improve safety and viability of one-time operation in the therapy of cleft lip and palate and cleft alveolar process. METHODS: Under the air-vein anesthesia, one-time operations were performed in 42 patients with complete cleft lip and palate plus cleft alveolar process. After rehabilitation in ICU, all patients were put into wards, and left hospital 4-7 days after operation. RESULTS: The situation during anesthesia and operation was good. There were no statistically significant changes in blood pressure, oxygen saturation of blood, heart rate and cardiac rhythm before, during and after operation. No anesthetic accident and no complication happened. The loss of blood was about 42.7 +/- 6.5 ml and the duration of operation was about 139.2 +/- 18.6 minutes. The rehabilitation was successful after operations. First intention took place in almost all patients' incisions, and palatal fistulas appeared in only 2 cases. CONCLUSION: Under the control of provisioned pediatric anesthesia and rehabilitation, one-time complete cleft lip and palate plus cleft alveolar process closure is safe and viable. PMID- 12539296 TI - [Diagnosis and treatment of central hemangioma of mandible]. AB - OBJECTIVE: To investigate methods of diagnosis and treatment of hemangiomas of the mandible. METHODS: Nineteen cases of central hemangioma of the mandible treated in our department from 1971 to 1997 were studied retrospectively. Fourteen patients received surgical treatment in which ten patients had received complete curettage of the lesion and the other four had received part resection of the mandible. RESULTS: The most common signs and symptoms of this lesion were hypermobility of teeth and episodes of spontaneous hemorrhage from the surrounding gingival sulcus. Selective angiography was significant in the diagnosis of this lesion. Fourteen patients received surgical treatment and thirteen lesions were removed completely, except that one died of massive bleeding at the operation and DIC on the next day. CONCLUSION: Selective angiography is useful to diagnosis the central hemangioma of mandible, and the complete curettage of the hemangiomas in the body of mandible is the most appropriate operation for children. At the same time, ligature of the inferior alveolar artery can reduce the risk of bleeding during operation. PMID- 12539297 TI - [An assessment of Helkimo index in 88 orthodontic patients before treatment]. AB - OBJECTIVE: The prevalence of temporomandibular joint disorders(TMJDs) was studied in orthodontic patients before orthodontic treatment. METHODS: The functional examination of the masticatory system was made with Helkimo index in 88 young patients with general malocclusion. RESULTS: According to the dysfunction index, 50 percent of the patients was free of anamnestic dysfunction, and 44.32% and 5.68% of the patients had a mild and a severe anamnestic dysfunction respectively, 32.95 percent of patients was free of clinical dysfunction, and 51.14% and 14.77% of patients had a mild and a moderate clinical dysfunction respectively, only 1.14 percent of patients had a severe clinical dysfunction. The dysfunction of masticatory muscles and sounds of temporomandibular joint were most common in the signs and symptoms of TMJDs. CONCLUSION: There were no significant differences in the distribution of Ai and Di in different gender and between Angle's Class I and II malocclusion, and there was a relationship between Ai and Di. PMID- 12539298 TI - [The efficacy on natural extrinsic stain of Colgate whitening dentifrice--eight week study]. AB - OBJECTIVE: To assess the effect on natural extrinsic dental stain by use of a dentifrice containing 1% tetrasodium polyphosphate (TSPP), 7% sodium tripolyphosphate (STPP), 0.243% sodium fluoride and 10% high cleaning silica (HCS) over an eight-week period was compared to a sodium fluoride/silica placebo dentifrice without stain removal ingredients. METHODS: Ninety one subjects were selected and stratified into two balanced groups randomly assigned to use the test dentifrice or the placebo dentifrice. The two groups were well balanced with regard to their mean baseline stain index scores, gender and tobacco habits. Subjects were instructed to brush their teeth twice daily (morning and evening) for one minute with their assigned dentifrice and a commercially available soft bristled toothbrush. Extrinsic dental stain examinations, which included mean stain area and mean stain intensity, were conducted at baseline, four and eight weeks. The same dental examiner conducted examinations on the subjects at each examination. RESULTS: After eight-week use of their assigned products, those subjects in the TSPP/STPP/HCS dentifrice group demonstrated a statistically significant (P = 0.0001) stain reduction of 37.1%, as compared to the fluoride placebo dentifrice group without stain removal ingredients. CONCLUSION: The use of a dentifrice containing 1% tetrasodium polyphosphate, 7% sodium tripolyphosphate, 0.243% sodium fluoride and 10% high cleaning silica is effective in reducing natural extrinsic dental stain, when compared to a placebo fluoride dentifrice. PMID- 12539299 TI - [Effects of muscular position splint on maxillomandibular realignment]. AB - OBJECTIVE: To study the effects of muscular position splint on maxillomandibular realignment. METHODS: The changes of occlusal impression on splint in 102 craniomandibular disorder (CMD) patients were recorded after wearing the splint 1 week, 2 weeks, 1 month, two to three or six months. If there appeared instability of occlusal impression when biting, the original impression was cleared and new soft resin was placed on its surface in order to get a new impression. The splint was always adjusted to fit well. When the signs and symptoms of CMD disappeared, the splint was worn off. Finally, whether the muscular contact position harmonized with the intercuspal position (ICP) or not was recorded. RESULTS: There appeared two different conditions: 1. The original occlusal impression remained unchanged in 66 patients; 2. Being unfitted in 36 patients, occlusal analysis revealed that 12/102 patients had abnormal occlusion including premature anterior teeth contact (2 cases), natural defective vertical dimension (1 case) and deflected ICP (9 cases). These abnormal occlusion could not be determined before treatment. CONCLUSION: The muscular position splint can correct the abnormal muscular position. On the premise that the muscular position is normal, the muscular position splint can be used to judge whether the ICP is normal. PMID- 12539300 TI - [A study of individual operation designs for unilateral cleft lip repair]. AB - OBJECTIVE: To formulate a reasonable and effective individual operation design that can be adaptable to various forms of deformity of unilateral cleft lip. METHODS: The unilateral deformity of unilateral cleft lip was classified into four types (Type I, II, III, IV) according to the distances from the midpoint of the base of columella and the points of alar bases respectively to cupid's bow peaks of both sides. Furthermore, according to the characteristics of deformity, four types of operation were designed respectively under the standard of making both cupid's bow peaks along cleft drop in the same levels, and the essentials were restoring and making the distances from the midpoint of the base of columella and the points of alar bases to both cupid's bow peaks symmetric. RESULTS: Type I operation was almost the same as the Millard's operation design, and the basic designs of Type II and Type IV operation were also the same as the Millard's operation design, but were combined with the Tennison's operation design. Type III operation was a new operation design of unilateral cleft lip repair. CONCLUSION: The individual operation design in unilateral cleft lip repair should be definitely according to the characteristics of cleft lip deformity. PMID- 12539301 TI - [A study of individual operation designs for bilateral cleft lip repair]. AB - OBJECTIVE: To design an individual operation procedure that can be adaptable to various deformity of bilateral cleft lip and to assure the symmetry of lip and cupid's bow postoperatively in bilateral cleft lip. METHODS: The distance from both sides of columella to cupid's bow peaks in prolabium as well as both sides of alar base to cupid's bow peaks in lateral lip has been used as standard of classification of the deformity of bilateral cleft lip, then operative procedures respectively was designed in accordance with each special characteristic of bilateral deformity. RESULTS: Five kinds of operation procedures were designed, and each procedure was suitable only for one kind of preoperation deformity of bilateral cleft lip. CONCLUSION: The individual operation designs should be corresponding to the types of deformity before operation in bilateral cleft lip repair. PMID- 12539302 TI - [The effects of fluoride on the bonding strength of brackets]. AB - OBJECTIVE: To evaluate the effect of using fluoride during and after etching on the bonding strength of brackets. METHODS: 45 extracted human premolar were divided into three groups: Group 1, control, not applying fluoride. Group 2, applying fluoride during etching. Group 3, applying fluoride after etching. In each group, the teeth were treated with etching and fluoride respectively and stored in water for 24 hours. Then brackets bonding strength were measured. RESULTS: The results showed that there was not significant difference among the three groups. CONCLUSION: Fluoride can be used to prevent enamel white spot. PMID- 12539303 TI - [Scanning electron microscopy study on different orthodontic methods of using fluoride in enamel white spot prevention]. AB - OBJECTIVE: To determine which method of topical fluoride application could make the enamel uptake more fluoride. METHODS: 12 extracted human premolars were divided into three groups of 4 teeth each: Group 1, control and not applying fluoride; Group 2, applying fluoride during etching; Group 3, applying fluoride after etching. In each group, the teeth were treated with etching and fluoride respectively. The specimens were then examined with scanning electron microscopy. RESULTS: The results showed that there were more reaction products on the enamel surface of Group 3. CONCLUSION: There was more fluoride uptake into the enamel treated with fluoride after etching. PMID- 12539304 TI - [A radioimmunologic assay of insulin in lateral pterygoid muscle after functional mandibular protrusion in rats]. AB - OBJECTIVE: To investigate the effect of insulin on the lateral pterygoid muscle after functional mandibular protrusion in rats. METHODS: Radioimmunoassay was used to examine the level of insulin in lateral pterygoid muscle quantitatively after functional mandibular protrusion in young growing rats. RESULTS: The amount of insulin in lateral pterygoid muscle in young growing rats enhanced after functional mandibular protrusion. CONCLUSION: Insulin plays an important role in adaptive remodeling of lateral pterygoid muscle after functional mandibular protrusion. PMID- 12539305 TI - [An eight-factor analysis of early skeletal Class II malocclusion using Frankel appliance]. AB - OBJECTIVE: To investigate the change of eight-factor after Frankel appliance treatment in skeletal class II malocclusions. METHODS: X-ray cephalometric film. RESULTS: Frankel appliance can be used to enhance the growth of ramus (Factor 2) and body (Factor 1) of the mandible, decrease the angle of cranial base (Factor 8) and made the maxilla rotate (Factor 4) anteriorly, which were benefit to the treatment of class II malocclusion. However, neither the vertical (Factor 5) and sagittal (Factor 3) growth of maxilla nor the height of posterior alveolar bone (Factor 6) can be inhibited by use of Frankel appliance. CONCLUSION: Frankel appliance is suitable to the treatment of class II patients with deficient mandible and almost normal maxilla, while Frankel appliance would rather not be used for patients with overgrown maxilla or vertical growth pattern. PMID- 12539306 TI - [Effects of BMP on periodontal ligament cell's cell cycling and DNA synthesis: a flow cytometry (FCM) analysis]. AB - OBJECTIVE: To investigate the effects of bone morphogenetic protein (BMP) on the cell cycling and DNA synthesis of periodontal ligament cells (PDLCs). METHODS: The 7th passage PDLCs were cultured in a 5% CO2, humid atmosphere at 37 degrees C in DMEM supplement with 10% fetal bovine serum (FBS) for 24 hours, then BMP was added in medium with 1% FBS. After being cultured another 72 hours, cells were trypsinized, washed and fixed with cold alcohol for 1 hour. To measure DNA content, 1.0 x 10(6) cells were washed with PBS, resuspended in 200 microliters PBS mixed with staining liquid (RNA ase, 0.1% triton-100 and propidium iodide) for 30 minutes at 4 degrees C. DNA specific fluorescence was measured on a Coulter Epics Profile-II (Coulter, USA) using 250 mW 488 nm laser light for excitating and collecting fluorescence at 610-630 nm. Phase fraction analysis was performed on DNA content histograms with multicycle software. RESULTS: When compared with the control, the percentage of PDLC in G1 during exponential growth was decreased by a function of BMP, while the percentage of S and (G2 + M) was increased in the experimental group. CONCLUSION: It suggested that the proliferation of PDLC be most probably regulated in the G1 phase of the cell cycle. BMP can be regarded as a positive regulatory growth factor. PMID- 12539307 TI - [Studies on standard growth curve chart on body weight of adolescent Chinese for orthodontic diagnosis]. AB - OBJECTIVE: To establish the first new annual increase and growth curve charts on the body weight of Chinese children. METHODS: Data were surveyed in 1995, which were obtained from subjects consisting of 32524 students aged from 7 to 19 years in Sichuan province. RESULTS: Mean values of the annual increment of body weight, sex and area difference were analyzed. CONCLUSION: The new standard growth curve chart on the body weight is very useful for orthodontic diagnosis, treatment and prognosis. PMID- 12539308 TI - [A dynamic study of influences of TMJOA on TMJ disc]. AB - OBJECTIVE: To learn the changes of TMJ disc positions under the influences of TMJ Osteoarthrosis(TMJOA). METHODS: The TMJOA experimental animal model was established by 12 goats. The TMJ disc positions were observed and recorded at different intervals, (1 month, 2 months, 3 months, 4 months and 6 months) then the pathological changes of TMJ specimens were also studied. RESULTS: Significant anteromedial shift of TMJ disc position was observed in some certain TMJOA phases and became more obviously along with the disease progress. CONCLUSION: The shift of TMJ disc may be the secondary change of TMJOA. PMID- 12539309 TI - [Expression of CDK4 protein in oral carcinogenesis]. AB - OBJECTIVE: To study the expression of cyclin dependent kinase 4(CDK4) protein in oral carcinogenesis. METHODS: An LSAB immunohistochemistry method was applied to 80 cases of paraffine-fixed tissues to study the expression and distribution of CDK4 in normal oral mucosa, oral premalignant lesion (OPL) and oral squamous cell carcinoma. RESULTS: The overexpression of CDK4 protein was observed in the progression of OPL. Meanwhile, a correlation analysis was applied to investigate the relationship between P16 and CDK4 expression, and the result showed that the coefficient of correlation was 0.813, which was statistically significant. CONCLUSION: The overexpression of CDK4 is involved in the progression of OPL, and there is a positive feedback loop between CDK4 and P16. PMID- 12539310 TI - [Partial gene clone and nif gene homologous sequence analysis of Streptococcus sanguis]. AB - OBJECTIVE: To analyze the sequence of Streptococcus sanguis chromosome which contains one DNA fragment of 800 base pairs (bp) and discuss Streptococcus sanguis biological features of heredity. METHODS: Streptococcus DNA of 800-bp genetic fragment was cloned and analyzed by using eukaryotic expression vector. RESULTS: By Genbank database, it showed that the 800-bp genetic sequence was highly homologous with other bacterial nifS and nifU gene, and the highest homologous score was 114. CONCLUSION: This nif gene of ATCC 10556 strain may correlate with nutrient metabolism and peroxide hydrogen release of Streptococcus sanguis. PMID- 12539311 TI - [Cloning and sequencing of variable region gene of heavy chain of monoclonal antibody against SA I/II of Streptococcus mutans]. AB - OBJECTIVE: To clone and sequence a immunoglobulin variable region of heavy chain (VH) from a mouse hybridoma 2B12F6, which produce monoclonal antibody against SA I/II of Streptococcus mutans. METHODS: The immunoglobulin variable region gene of heavy chain of 2B12F6 was amplified and cloned into pUC18 by using PCR technique and gene engineering technique, and then the gene sequence was analyzed by Sanger's method. RESULTS: The VH gene segment was 360 base pairs in length and coded 118 amino acids, and the homology of framework of VH gene and mouse VH gene published was 70%, which accorded with the feature of mouse VH gene. CONCLUSION: The VH gene gained from 2B12F6 could provide the possibility of construction of gene engineering antibody against SA I/II of Streptococcus mutans. PMID- 12539312 TI - [Effects of hexamethylene bisacetamide on P53 protein expression of MEC-1 cell line and cell differentiation]. AB - OBJECTIVE: To investigate the relationship between the differentiation of the mucoepidermoid carcinoma cell line MEC-1 induced by HMBA and the expression of P53 protein. METHODS: Effects of HMBA on the growth of MEC-1 cells in vitro were measured with MTT colorimetric assay, and the content of DNA in cells was assayed by using Feulgen's staining. The P53 protein expression of MEC-1 cells was detected with ABC immunohistochemistry with anti-mutant P53 protein antibody, and the distribution of cells in different differentiation phase was determined by flow cytometry. RESULTS: HMBA inhibited the growth of MEC-1 cells in a dose dependent and time-dependent manner, and HMBA-treated MEC-1 cells were arrested in the G1 phase and had a lower content of DNA and a lower level of P53 protein compared with the control group. CONCLUSION: The differentiation of MEC-1 cells induced by HMBA may be associated with the regulation of HMBA on the P53 protein expression and the cell differentiation cycle. PMID- 12539313 TI - [An experimental study on bone potentials at fracture site with axial interfragmental gradient compression]. AB - OBJECTIVE: To evaluate the mechanism of stress generated potentials in compressive fixation leading to the primary healing. METHODS: Eighteen fresh sheep tibiae were divided equally into three groups, and all specimen were fractured artificially and imposed "prestressing" at fracture sites with tension device and plate, meanwhile stainless wire tension band was used in the place opposite to plate. The interfragmental pressure was exerted when applied plates, which were 98.62 +/- 19.74 N, 137.93 +/- 8.13 N and 169.20 +/- 11.33 N. Bone potentials at fracture sites were recorded with reference to the potential measurement system by Friedenberg and Brighton (1966). RESULTS: There was no significant statistical difference in bone potentials when made comparison between before and after compression fixation in each group and among groups of different axial interfragmental gradient compressions. CONCLUSION: The mechanism of stress generated potentials may not exist in primary healing with compressive fixation. PMID- 12539314 TI - [Effects of alumina particle-size on mechanical properties of alumina-glass composite]. AB - OBJECTIVE: To study the effects of alumina particle-size on mechanical properties of alumina-glass composite. METHODS: Alumina compacts were prepared by isostatic press method and sintered at 1400 degrees C for 2 hours with two various particle size alumina. Alumina-glass composite was made by infiltrating partially sintered porous alumina with glass at 1200 degrees C for 4 hours. RESULTS: The strength and fracture toughness of nanometre alumina-glass composite were 358.2 MPa, 2.38 MPam1/2 respectively. The strength and fracture toughness of micrometer alumina glass composite were 432.2 MPa, 5.12 MPam1/2 respectively. Image analysis of SEM micrograph showed that porous rate was about 11.5% in the nanometer alumina-glass composite, 1.2% in the micrometer alumina-glass composite, and there was more glass composition in the nanometer alumina-glass composite than that in the micrometer alumina-glass composite. CONCLUSION: The alumina volume percentage and porous rate play an important role in the strength and fracture toughness of alumina-glass composite. PMID- 12539315 TI - [Study on the crystal heat treatment schedule of Plat-II castable ceramic]. AB - OBJECTIVE: To establish the crystal heating schedule of Plat-II castable ceramic materials. METHODS: This study first identified the thermal limits of Plat-II castable ceramic crystallization basing on DTA exotherm analysis, then the crystal density of various heating schedule was observed by scanning electron microscope. RESULTS: The efficient crystal heating schedule of Plat-II castable ceramic was established: The temperature was increased from the room temperature to 620 degrees C with the speed of 10 degrees C/min, kept 20 min, then was increased continually to 670 degrees C with the speed of 5 degrees C/min, and kept 30 min. CONCLUSION: The establishment of the crystal heat treatment schedule of Plat-II castable ceramic will become the basis for the further study. PMID- 12539316 TI - [The boundary design of mandibular model by means of the three-dimensional finite element method]. AB - OBJECTIVE: To construct the mandibular model and simulate the boundary conditions by means of the three-dimensional finite element method. METHODS: A juvenile mandible, including TMJ, was scanned transversely by CT, then a three-dimensional finite element model of mandible was constructed. At the same time, the functions of stomatologic muscles and ligaments and the functional actions of temporomandibular joint (TMJ) were simulated by means of cable elements and compressive gap elements respectively. RESULTS: The cable elements were consistent with the functions of stomatologic muscles and ligaments, and the gap elements were similar to the boundary conditions of the TMJ and teeth. CONCLUSION: Cable and gap elements are reasonable boundary conditions of the finite element model of mandible, which will improve the similarity of the model. PMID- 12539317 TI - [Animal experiment of the ketamine effects on traumatic brain injury resulting from impact maxillofacial injury]. AB - OBJECTIVE: To test the effects of ketamine (KET) on traumatic brain injury resulting from impact maxillofacial injury. METHODS: The rat models with middle serious traumatic brain injury were obtained by falling impaction at maxillofacial region of the rats, then the animals were treated with KET in the early stage, and the alteration of the neural function in those rats was observed at the same time. RESULTS: There was obvious difference between the rats in the KET-treating groups and the control group. It took about 2 days fewer than that of the control group for rats in the KET-treating group to recover. CONCLUSION: KET can shorten the recovery time of rats with brain injury resulting from impact maxillofacial injury. PMID- 12539318 TI - [Psychological aspects of patients with burning mouth syndrome: an investigation of personality, psychopathological profiles and social life background stress]. AB - OBJECTIVE: To know the psychosocial background of patients with burning mouth syndrome (BMS) in China. METHODS: In this study, 30 patients with BMS and 30 normal subjects of control group were evaluated for their personality, psychopathological profile and existence of recent life events. All subjects were required to complete the EPQ questionnaire, the SCL-90 questionnaire and a Life Event Scale, and chi-square test analysis of variance and rank sum test were used in this study. RESULTS: The personality of introversion and instability was common in BMS group. The scores of 9 emotional factors and negative life events were higher than that of the control group significantly. CONCLUSION: BMS may be a series of psychological disorder and somatic discomfort caused by stresses of social life events on the people with special personality. PMID- 12539319 TI - [Clinical observation of dental pulp and root changes of juvenile anterior permanent teeth after concussion and luxation]. AB - OBJECTIVE: To observe the incidence of pulp necrosis and root resorption of the juvenile anterior permanent teeth with concussion and luxation injuries and analyze the relationship between the type of injury and form of root resorption. METHODS: 157 teeth of 79 patients were chosen from 201 patients with concussion and luxation treated with functional-bite-plate at the Department of Pedodontics, School of Stomatology, Beijing Medical University from 1984 to 1996. RESULTS: High occurrence of pulp necrosis and root resorption were found in teeth with intrusion, and pulp necrosis occurred most within 3 months after the injury, while root resorption occurred during 3 weeks to 2 years. At the same time, surface resorption occurred mainly in the teeth with intrusion, extrusion and lateral luxation, and replacement resorption occurred only in intrusive teeth, however inflammatory resorption occurred in extrusive teeth whose pulp was infected. CONCLUSION: Intrusion is the most severe type of tooth trauma. PMID- 12539320 TI - [Atraumatic restorative treatment fillings and fissure sealants in permanent teeth--a 2-year study]. AB - OBJECTIVE: To study the feasibility of providing Atraumatic Restorative Treatment (ART) in schools and in rural areas in China, and to evaluate the acceptance and effectiveness of the treatment. METHODS: The study was conducted among 272 grade one students chosen from four different secondary schools in Deyang of Sichuan Province, China. The mean age of the subjects was 12.5 years. 107 boys and 165 girls were found at the baseline examination to have teeth that were suitable for receiving ART fillings or sealants. The treatments were performed within a month using the glass ionomer Ketac-Molar manufactured by ESPE. Evaluation was carried out at 3 months, one year and two years after treatment. RESULTS: ART fillings were placed in 295 permanent teeth, and the 1-year and 2-year success rates were 95.7% and 91.1% respectively. ART sealants were placed in another 191 permanent teeth, and the 3-month, 1-year and 2-year success rates were 95.2%, 89.1% and 78.8% respectively. The incidence of recurrent caries was very low after two years, which were 2.1% and 0.6% for the fillings and sealants respectively. ART was accepted by the majority of the students. CONCLUSION: ART is a simple, acceptable technique, and is suitable for widespread adoption in China. PMID- 12539321 TI - [Analysis of 413 cases of mandibular fractures]. AB - OBJECTIVE: To investigate the characteristics of mandibular fracture. METHODS: 413 patients with mandibular fracture were chosen from patients who received treatment at the department of oral and maxillofacial surgery, College of Stomatology, West China University of Medical Sciences from 1986 to 1995, and a retrospective study was carried out. Chi-square test, t-test and linear regression were used to analyzed age, sex, causes, anatomic site, complicated injury, and treatment of jaw fracture. RESULTS: Fracture of Mandible occurred mainly in male, and during 20 to 29 years old. Mental fracture was common in mandibular fracture, and fracture of central craniofacial bone and craniocerebral injury were often complicated with jaw fracture. CONCLUSION: Mandibular fractures are caused mainly by traffic ever since 1990, and rigid intrafixation is a satisfactory treatment which has been accepted by surgeons. PMID- 12539322 TI - [Two-dimensional finite element analysis for morphology of craniofacial hard tissue of people with anterior crossbite at early permanent dentition]. AB - OBJECTIVE: To analyze the morphology of craniofacial hard tissue of people with anterior crossbite at early at early permanent dentition. METHODS: The craniofacial morphology of hard tissue of 80 cases anterior crossbite of the early permanent dentition was analyzed by a special computer software system for finite element analysis of two-dimensional cranio-facial structure. RESULTS: The size change of each element was smaller than shape change in cross-bite group, which suggested that the shape change was the main element change; The posterior cranial base was found abnormal in both male and female groups, with counterclockwise rotation occurring at posterior cranial base to some extent; The upper part of face and the maxillary suffered underdevelopment in both male and female groups; The most outstanding change in size and shape occurred at anterior and superior part of mental, with each component of counterclockwise rotation to varying degree. CONCLUSION: Abnormality that occurs in craniofacial hard tissue of people with anterior crossbite at early permanent dentition period is highlighted by abnormal skeletal shape change and rotation, and may be one of the fundamental factors causing crossbite. PMID- 12539323 TI - [Clinical therapeutic effects observation of iodophors on irrigation and disinfection of root canal]. AB - OBJECTIVE: To evaluate clinical effects of iodophors on root canal therapy. METHODS: 147 teeth with root canal therapy collected from clinical patients were divided into two groups: experimental and control group. In experimental group, 0.05% and 0.5% iodophors were taken as irrigate agent and disinfectant, respectively, while in control group, 3% H2O2 + 0.9% NS and Fc were used. RESULTS: Clinical therapeutic effects evaluation indicated that there was no statistically significant difference of alteration of clinical syndromes after the administration and the therapeutic effect between experimental group and control group during the period of root canal therapy. CONCLUSION: Iodophors could be taken as a kind of candidate drugs on irrigation and disinfection of root canal therapy. PMID- 12539324 TI - [Morphologic characteristics of dental cuticle on teeth affected by diabetic periodontitis]. AB - OBJECTIVE: To determine the nature of the dental cuticle found on teeth with diabetic periodontitis. METHODS: Thirteen permanent teeth which were extracted from three diabetic patients with periodontitis were studied, with the control group of 16 healthy teeth extracted for orthodontical reasons, then all the teeth were decalcified with 5% nitric acid, stained with HE, and examinated by using light microscope. RESULTS: A layer of thick coat, dental cuticle, covered the root surface of the teeth with diabetic periodontitis, and the cuticle which presented regular laminar structure extended from the cementoenamel junction to the apical portion with different thickness from 40 to 80 microns, but no such similar cuticle structure was found on the healthy teeth of the control group. CONCLUSION: There is abnormal cuticle structure on the teeth of diabetic patients with periodontitis. PMID- 12539325 TI - [Cloning and sequence characterization of heavy chain variable region gene from hybridoma cells secreting MAB against VEGF]. AB - OBJECTIVE: To clone the variable region of heavy chain (VH) gene from hybridoma secreting monoclonal antibody against human vasoendothelial growth factor (VEGF) for construction of recombinant antibody (scFv). METHODS: One step method was used to extract total RNA and degeneration primer was used to amplify the cDNAs by retropolymerase chain reaction (RT-PCR), then the products were inserted to T vector and its sequence was analysed. RESULTS: The VH gene contained 369 base pairs encoding 123 amine acid resides. Analysis of the nucleotide sequence of the light chain variable region revealed that the VH sequence was 87% identical to the sequence present in mouse Ig VH region. CONCLUSION: The VH gene was 369 bp. According to Kabat classification, the VH gene belonging to mouse kappa light chains subgroup I (A), resulting from VH-D-JH4 rearrangment. PMID- 12539326 TI - [Design and synthesizing of human vascular endothelial growth factor (VEGF) peptide]. AB - OBJECTIVE: To predict continuous epitopes of VEGF using synthetic peptide of VEGF as antigen to produce anti-VEGF-peptide monoclonal antibody. METHODS: According to amino sequence of VEGF189, antigen index program and Goldkey soft were used to predict VEGF epitopes, and the peptide containing one of predicted epitopes was synthesized by 430A Peptide Synthesizer. RESULTS: The NH2-terminal 1-26 amino acid sequence of VEGF was determined as synthetic peptide and synthesized. CONCLUSION: The predicted epitope of VEGF was synthesized, which provided an effective method when lacking of VEGF. PMID- 12539327 TI - [Internal fixation for zygomatic arch fracture with super-high molecular weight poly D, L-lactic acid mini-plates and screws: a study in dogs]. AB - OBJECTIVE: To investigate the internal fixation effects of super high molecular weight (Mv = 6.0 x 10(5) kD) poly D, L-lactic acid(PDLLA) mini-plates and screws. METHODS: 10 dogs were utilized in this experiment with the self-contrast study method, comparing with the titanium mini-plates and screws. RESULTS: The mechanical properties of the PDLLA mini-plates and screws appeared to be sufficient to enable undisturbed healing of depressed zygomatic arch fracture. CONCLUSION: The good fixation effect of PDLLA mini-plates and screws was as same as that of the titanium mini-plates and screws without the need of secondary operation. PMID- 12539328 TI - [Tissue reaction and degradation of super-high molecular weight poly D, L-lactic acid mini-plates and screws: an animal experiment]. AB - OBJECTIVE: To investigate the tissue reaction and degradation processes of the super-high molecular weight(Mv = 6.0 x 10(5) kD) poly D, L-lactic acid(PDLLA) mini-plates and screws through the internal fixation for zygomatic arch fracture. METHODS: It was carried out by the self-contrast study method and comparing with the titanium mini-plates and screws. RESULTS: There was no side-effects on bone healing. The reaction of soft and hard tissues to PDLLA mini-plates and screws was similar to that of the titanium mini-plates and screws. No complication had been seen with the use of PDLLA mini-plates and screws, including infection, foreign body reaction and underlying osteolysis. The PDLLA devices had no changes in shape after 3 months, and then became the grains in different sizes after 6 months, and were completely resorbed within 12 months in vivo. CONCLUSION: The super-high molecular weight PDLLA mini-plates and screws has good biocompatibility and proper degradation time. It is safe and effective to use the mini-plates and screws made of the super-high molecular weight PDLLA for internal fixation of bone. PMID- 12539329 TI - [A study on the biological properties of fimbriae of Actinomyces viscosus. I. The preparation and determination of fimbriae of A. viscosus]. AB - OBJECTIVE: To build up the methods to prepare and determine the fimbriae of A. viscosus. METHODS: Type 1 and 2 fimbriae were prepared from A. viscosus 5519 and 5951 respectively. Fimbriae were isolated from bacteria cells by mechanical shaking, obtained by collecting the supernatant after centrifugation, and purified by ammonium sulfate precipitations and gel filtration. RESULTS: Electron microscope showed that fimbriae existed in the preparations. The immunological reaction between the fimbriae preparations and specific antibodies against fimbriae attested that they were type 1 or type 2 fimbriae. CONCLUSION: The present study become a foundation for further study on the biological properties of fimbriae of A. viscosus. PMID- 12539330 TI - [Determination of major expansion properties of refractory die material compatible with slip casting core of sintered titanium powder]. AB - OBJECTIVE: To determinate major expansion properties of refractory die material. METHODS: The setting expansion ratio of refractory die material for slip casting core of sintered titanium powder at room temperature was performed, as well as thermal expansion ratio from room temperature to 800 degrees C. RESULTS: The maximum setting expansion ratio in 2 hours reached 0.3407%; The final setting expansion ratio in 24 hours was 0.3117%; The mean thermal expansion coefficient was mainly in range of 8 x 10(-6)-11 x 10(-6)/degree C; The expansion property seemed very stable after sintering repeatedly and the small shrinkage after sintering could be compensated with the die spacer and setting expansion. CONCLUSION: The expansion properties of the refractory die material that we synthesized can fulfil the application requirements of slip casting core of sintered titanium powder. PMID- 12539331 TI - [A comparative study of mechanical properties of microwave-cured and conventional water bath-cured denture base resins]. AB - OBJECTIVE: To study the mechanical properties of microwave-cured polymethyl methacrylate (PMMA) denture base resins. METHODS: Using experiment research, the mechanical properties of microwave-cured and water bath-cured PMMA materials were compared. RESULTS: The microwave-cured acrylic strips had equal or better mechanical properties than conventionally processed strips. CONCLUSION: The microwave-cured denture materials satisfies the requirements of the prosthetic clinic. PMID- 12539332 TI - [Effects of two growth factors combination on the proliferation and differentiation of human osteoblast-like cells]. AB - OBJECTIVE: To examine the effects of platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF-II) and type beta transforming growth factor(TGF-beta) which were believed to have biological effects on bone cells on human osteoblast-like cells. METHODS: 3H-TdR, 3H-Proline and alkaline phosphatase of the osteoblast-like cells were tested to study the combinating effects of different growth factors on the proliferation and differentiation of osteoblast-like cells. RESULTS: The combination of different growth factors had promotive effect and synergic interact on proliferation and differentiation of osteoblast-like cells. CONCLUSION: The phenomena indicate that the combination of two growth factors will enhance the biological effects of bone formation. PMID- 12539333 TI - [Fibronectin in adhesion, spreading and proliferation of mandibular condylar cartilage cells on cytodex-3 microcarrier]. AB - OBJECTIVE: The purpose of this study was to investigate the effects of exogenous fibronectin on the attachment and growth of mandibular condylar cartilage (MCC) cells on DEAE-dextran micro-carrier. METHODS: MCC cells were harvested from newborn New Zealand white rabbits by sequential digestion with trypsin and collagenase. They were grown on the 20 mg/L fibronectin-coated and uncoated cytodex-3 micro-carrier. Samples were collected after 1, 3, 5, 7, 9, 11 days. The kinetics of adhesion and growth were observed with phase contrast microscope and environmental scanning electronic microscope (ESEM) and quantified by 0.1% crystal violet nuclear extrusion method. RESULTS: MCC cells could rapidly attach and spread onto the fibronectin-coated cytodex-3, and nearly 60 percent cells attached to the micro-carrier within 2 hours. Spreading cells were flat and lowly refractile observed under phase contrast microscope. While the attachment and spreading of the cells in the untreated group were slow, and only 36 percent cells attached to the uncoated micro-carrier. However, most cells attached to the micro-carrier in both groups after 24 hours. The MCC cells attaching to FN-coated cytodex-3 observed under ESEM were flatter and adhered tightly with more processes (pseudopodia) stretching out from cytoplasm. In contrast, cells on untreated cytodex-3 remained regularly round even 24 hours after being plated. Meanwhile, there was an accelerated growth rate of the MCC cells on the fibronectin treated cytodex-3. The cell density in the FN treated group was higher than that of the untreated group at the end of the culture. CONCLUSION: Fibronectin, one major glycoprotein of extracellular matrix, plays an important role in cell attachment and spreading. The better understanding of the extracellular matrix will be helpful in selecting the optimal substrate for cell growth in vitro. PMID- 12539334 TI - [Expression of type X collagen in condylar cartilage during mandibular protrusion]. AB - OBJECTIVE: This study was designed to detect type X collagen expression, at both gene and protein levels, in condylar cartilage during mandibular protrusion triggered by functional appliance. METHODS: One hundred SD rats with age of 5 weeks were selected and divided into 5 experimental groups (n = 5) and 5 control groups (n = 5). Animals in experimental groups were imposed with bite-jumping appliances which caused identical magnitude of mandibular protrusion. The animals of experimental groups, together with their matched controls, were sacrificed 3, 7, 14, 21 and 30 days after the appliance insertion. 6 microns sections were cut from TMJ through the sagittal plane. In situ hybridization and immunohistochemical analysis were performed to detect the expression of type X collegen mRNA transcription and its protein synthesis. RESULTS: 1. Intensive staining of type X collegen mRNA was found to distribute within the whole area of hypertrophic zone, a sharp contrast to the control groups which showed very little or no staining. 2. Type X collegen protein synthesis, which was highlighted by immunofluoscent staining, was found to spread over the whole hypertrophic zone, compared to the controls that demonstrated a very little staining. CONCLUSION: Expression of type X collagen in condylar cartilage indicates the onset of endochondral ossification in condyle in response to mandibular protrusion provoked by functional appliances. PMID- 12539336 TI - [Tensile mechanics of mandibular condylar cartilage]. AB - OBJECTIVE: To study the biomechanical properties of the mandibular condyle cartilage and determine its functional importance. METHODS: Eight condyle specimens of the four pig temporomandibular joints were obtained by autopsy and stored in sealed plastic bags at -20 degrees C. These pigs were healthy, aged 1 year +/- 2 months. In the coronal direction, 300 microns wide, rectangular cartilage-bone specimens were prepared on the right condyles using specified knife. In the sagittal direction, rectangular cartilage-bone specimens with the same width were obtained on the left condyles. Standard rectangular testing specimens were prepared from cartilage-bone specimens in a microtome kept at -20 degrees C (300 microns wide and 250 microns thick). The specimens were inserted in a soft-tissue uniaxial tensile test machine and immersed in a bath of Ringer's solution at room temperature. Preconditioning of the specimens was carried out prior to the uniaxial tensile experiment and the specimens were elongated to failure with the constant strain rate (0.05 mm/s). The resulting mean values were curved fitted with Fung's two-parameter (A, B) exponential stress strain equation using a nonlinear regression program. RESULTS: The data obtained showed that a typical nonlinear biomechanical behavior of the condyle cartilage. Toe-region and quasi-linear zone were obvious on the experimental stress strain relation curve. Theoretically fitted parameter A and B were 0.47 and 4.13 (coronally) and 0.60 and 4.70 (sagittally). The sagittal specimens showed greater tensile stress (2.92 MPa), tensile stiffness (9.04 MPa), energy absorption (6.02 N.mm) but less tensile strain (37.34%) than that of the coronal specimens (2.15 MPa, 6.55 MPa, 4.91 N.mm, and 41.12%). The differences were statistically significant (P < 0.05). CONCLUSION: The mandibular condyle cartilage is a kind of biophasic material which shows nonlinear viscoelastic behavior, anistrophic characteristics in biomechanical function and high ability to resist tensile stress in the longitudinal direction. PMID- 12539335 TI - [Immunohistochemical study of type I and type II collagen in the remodelling of the supraosseous tissue of mandibular condyle]. AB - OBJECTIVE: Previous studies focused on the pathological changes of the supraosseous tissue of mandibular condyle (STMC), but little is known about extracellular matrix during the remodelling activity of temporomandibular joint (TMJ). This study will examine the characteristics of type I and type II collagen during this process. METHODS: The animal model was established by extracting the left lower molars of adult rabbits. Temporomandibular joints of two sides were taken after 2 weeks, 1 month and 3 months respectively, and embedded in paraffin. Then 5 microns-thick sections were cut and processed for immunohistochemistry and haematoxylin-eosin staining. An immunohistochemical ABC method was used to determine the local changes of type I and type II collagen. The positive cells were counted in order to analyze quantitative changes. RESULTS: After loss of teeth from 2 weeks to 3 months, the number and arrangement of chondrocytes changed. Some chondrocyte clusters were also observed in the non-extraction side of TMJ. Most STMC cells stained positively with anti-type I collagen, but weakly in chondrocytes and hypertrophic chondrocytes. On the other hand, type II collagen only existed in chondrocytes and some hypertrophic chondrocytes. The expression of type I collagen decreased after 2 weeks and became stronger after 1 and 3 months, furthermore distributed unevenly in STMC. Some hypertrophic chondrocytes in the non-extraction side showed strong positive staining with anti type I collagen. Type II collagen in two sides decreased after 2 weeks, then became stronger after 1 and 3 month, but the whole level was less than that of the control group. Expressions of two kinds of collagen were compared between two sides. Type I collagen in the non-extraction side was stronger than that in the extraction side, while type II collagen had an opposite result. CONCLUSION: Adult rabbits have a limited remodelling ability, and the synthesis and secretion of type I and type II collagen will change when the remodelling is beyond a proper extent, which will lead to impaired resistance and elasticity of fibrocartilage. PMID- 12539337 TI - [Bax protein expression in the carcinogenesis of human oral mucosa]. AB - OBJECTIVE: Bax gene is an important apoptosis-promoting gene. In order to investigate the expression of Bax in oral premalignant lesions and oral squamous cell carcinomas, a total of 38 samples are evaluated using a labelled streptavidin-biotin (LSAB) immunohistochemical assay. METHODS: A total of 38 specimens were studied, including normal oral mucosa, premalignant lesions and squamous cell carcinomas. The specimens were obtained and blocked, fixed with 10% buffered formalin and embedded in paraffin using conventional histopathological techniques. 3 microns-thick sections of paraffin-embedded tissues were cut, mounted onto slides coated with 5% APES (3-aminopropyltriethoxysilane), dried overnight at 56 degrees C, dewaxed in xylene and rehydrated through descending graded alchols to phosphate-buffered saline (PBS, pH 7.4). For antigen retrieval, slides were immersed in 10 mmol/L sodium citrate buffer (pH 6.0) and boiled twice for 5 min in a microwave oven (800 W). After treated for 10 min with 3% H2O2, 18% methanol in PBS, the slides were covered with 10% normal porcine serum for 10 min at 37 degrees C. Then slides were incubated with primary antibody (bax rabbit polyclonal antibody) for 60 min at 37 degrees C and were subsequently incubated with prediluted biotinylated antibody against rabbit immunogobulins, and streptavidinhorseardish peroxidase conjugate for 30 min at 37 degrees C. After washing, peroxidase activity was detected using 3,3'-diaminobenzidine as chromogen with H2O2 as substrate. The cells in the test specimens which demonstrated granular staining were considered as positive. A haemocytometer counter with 6. 6 framework were applied and only the positive cells on the cross were counted. The cell counting were processed in ten randomly chosen 400* microscopic fields and the mathematic mean was presented as the final counting of each sample. Statistical valuations were performed using the version 6.0 SPSS package. Positive controls were sections of bladder cancer tissues. Blank controls were fabricated for each specimen by the omission of the primary antibody, which was replaced with PBS. RESULTS: In the process of oral carcinogenesis, each stage had Bax expression. The positive staining appeared in cytoplasm. In the normal oral mucosal specimens Bax expression was evident in the prickle layer, but not in the basal cell layers. Various degrees of Bax expression were seen in the diseased tissues. The staining pattern of hyperkeratotic lesions was similar to the normal oral mucosa, but Bax expression were also seen in the basal cell layer. In the mild, moderate, severe dysplasia and squamous cell carcinomas, Bax expression were seen in all layers, however, the intensity of staining were greater in mild and moderate dysplasia. The number of positive cells tended to increase gradually with the development of cell malignancy in the tissues of hyperkeratosis, mild and moderate dysplasia (P < 0.05). In the tissue of squamous cell carcinomas the number of positive cells had no marked difference comparing with the normal oral mucosa. CONCLUSION: The expression of Bax is involved in oral carcinogenesis and the compensative increase of Bax protein expression may be an early response. PMID- 12539338 TI - [The expression and mechanism study of BMP in pleomorphic adenoma]. AB - OBJECTIVE: One important pathological character of pleomorphic adenomas is that there is chorndriod component in the epithelial tumors. It is well known that bone morphogenetic protein (BMPs) plays important roles in the morphogenesis of hard tissue by inducing mesenchymal cells to differentiate into osteoblasts and chondroblasts in vivo. So we examined the expression of BMP and BMP2 mRNA in pleomorphic adenomas to study the pathological mechanism of chorndriod tissue formation. METHODS: Using ABC immunohistochemistry method, 6 specimens of pleomorphic adenomas were examined with BMP antibody. Reverse transcriptase polymerase chain reaction (RT-PCR) methods were developed for detecting BMP2 mRNA expression with special primers used to amplify the BMP2 cDNA mature domain fragment. The total RNA was extracted from frozen 6 specimens of pleomorphic adenoma and then reversed to cDNA by reverse transcription using AMV reverse transcriptase. PCR conditions were 94 degrees C for 20 seconds, 60 degrees C for 30 seconds, and 72 degrees C for 1 minute, 40 cycles, followed by extension for 30 minute at 72 degrees C. The reaction products were analyzed by electrophoresis. RESULTS: BMP expression could be found in all of the specimens of pleomorphic adenoma examined by immunohistochemistry method. BMP2 RT-PCR product was detected in only 5 specimens that showed BMP2 mRNA existed in 5 specimens. CONCLUSION: These results may be helpful to study the mechanism of pleomorphic adenoma. It is speculated that BMP secreted by tumor cells may play important roles in origin of osteoid tissue in pleomorphic adenoma by inducing neoplastic myoepithelial cells to differentiate to chondrocytes. PMID- 12539339 TI - [The experimental study on the activity of rhBMP-2, coral and collagen composites inducing intramuscle bone]. AB - OBJECTIVE: To investigate the ectopic bone induction activity of the composite of coral, collagen and rhBMP-2, and the degradation of the carriers of rhBMP-2. METHODS: The composites of rhBMP-2, coral and collagen were implanted into the muscle pouches of mice. The mice were divided into the following four groups. Group H: each composite had 2 mg rhBMP-2, Group L: each composite had 0.5 mg rhBMP-2, Group J: each composite had only type I collagen, Group C: only coral was implanted in the muscle of mouse. Each group had 20 mice. Every 5 mice in each group were killed postoperatively after 1, 2, 4 and 8 weeks. The ectopic osteoinductive activity was assessed with histology and histomorphometry. The point counting method was applied to measure the formation of new bone and cartilage and the degradation of coral in the composites. The data was analyzed statistically using t test. RESULTS: There was no obvious morbidity in any of the animals in this study, and no wound infection, bleeding, or feeding difficulties postoperatively. The samples were examined histologically. Cartilage and bone induction occurred in Group H and L. After 1 Week, new bone was observed in Group H, and cartilage formation occurred mainly in Group L. After 2 weeks, both cancellous bone and bone marrow elements were present in Group H and L. New bone formation was not uniform throughout the specimen. Part of the coral in the composites was not absorbed between 2 to 4 weeks postoperatively. At the 8th week, the coral in the composite was absorbed completely. While in Group J and C there was not any new cartilage or bone formation. According to the statistical analysis, bone formation occurred 2 weeks earlier in Group H than in Group L after operation. From 4 postoperative weeks, no difference of the bone formation rate was observed between Group H and Group L. The degradation rate of coral in Group C was the fastest among the four groups. CONCLUSION: The composites of rhBMP-2, coral and collagen are degradable bone substitutes with bone induction and conduction, and induce dosedependent and time-dependent amounts of intramuscle bone in mice during limited postoperative period. The coral in the composites could be absorbed completely. PMID- 12539340 TI - [Measurement of content of collagen type IV and laminin in tissue of oral squamous cell carcinoma and its clinical significance]. AB - OBJECTIVE: To investigate the relationship between the content of collagen type IV (ColIV) and laminin (LN) in tissue of oral squamous cell carcinoma and the clinicopathological parameters. METHODS: 30 patients with oral squamous cell carcinoma came into the study, 6 normal mucosa tissue came from patients who underwent orthogathic operations. The extraction solution was prepared by homogenizing of cancer and normal mucosa tissue in extraction buffer. The content of ColIV and LN in tissue of oral squamous cell carcinoma were measured by radioimmunoassay and represented by amount of per milliliter extraction solution. Mode of invasion (MI) was divided into 4 types basing on the relationship between normal and cancer tissue. The results of measurement of content of ColIV and LN were expressed using mean +/- s, whose correlation with clinicopathological parameters was evaluated by using Student t test. RESULTS: The MI classification showed that type I consisted of 4 cases, and type II comprised 12 cases. 5 and 9 cases belonged to type III and type IV respectively. The content of LN and ColIV in tumor tissue were 1.02 +/- 0.11 micrograms/ml and 1.83 +/- 0.21 micrograms/ml respectively, and the content of LN and ColIV in normal tissue were 1.98 +/- 0.23 micrograms/ml and 2.87 +/- 0.45 micrograms/ml respectively. There was significant difference between the content of ColIV and LN in normal and tumor tissue, and the content of ColIV and LN in normal tissue was much higher than that in cancer tissue (P < 0.05, P < 0.01). No relationship was found between the variation of ColIV and LN content and location and pathological grade of squamous cell carcinoma (P > 0.05), but it was found the content of LN and ColIV in patients with neck lymph node metastasis were greater than that in patients without metastasis (P < 0.05). Content of LN and ColIV also showed negative relationship with the MI index. CONCLUSION: The assay of LN and ColIV in tumor might be useful for diagnosis of metastasis of oral squamous cell carcinoma, and moreover, the relationship between ColIV and LN and MI also gives the suggestion that ColIV and LN may play some roles in preventing the invasion of oral squamous cell carcinoma. PMID- 12539341 TI - [A treatment combined Chinese with Western medicine for microcirculation diseases in tongue]. AB - OBJECTIVE: Glossodynia is a common oral disease. The purpose of the investigation is to analyse the mechanism of microcirculation disorder in glossodynia, and find a new effective treatment. METHODS: 18 cases with microcirculation disorder in tongue were studied by histopathology, hemorrheology and nail microcirculation, treated by intravenous injection of angelica for promoting blood circulation and removing blood stasis. RESULTS: Hyper-viscosity of blood was found in this group of patients. Abnormal micro-vessels were observed in microcirculation examination, though vasculitis wasn't detected in tongue tissue. CONCLUSION: Microcirculation disorder may play an important role in the cause of glossodynia. For elderly patients, cardiovascular and mental diseases should be considered. According to the theory of the blood stasis in chinese medicine, these studies show that a treatment combined chinese with western medicine can be applied for the patients, and injection of angelica can improve microcirculation. PMID- 12539342 TI - [The immunohistochemical and electron microscopical study of pathological classification of Melkersson-Rosenthal syndrome]. AB - OBJECTIVE: To prove further the pathological classification of Melkersson Rosenthal syndrome (MRS) and observe the morphological features of the granuloma cells in MRS. METHODS: We studied 20 cases of MRS, with 10 cases of granuloma type and the other 10 cases of non-granuloma type. The negative results of periodic acid-schiff (PAS), Neelren anti-acid special staining and tuberculosis (TB)-DNA polymerase chain reaction proved that all cases were not TB and fungous disease. Then all the cases were labelled with antibody of lysozyme by immunohistochemical staining and investigated with electron microscope. We studied the expression of lysozyme in multinucleated giant cells, epithelioid cells and histiocytes. RESULTS: The results of lysozyme staining were positive in the multinucleated giant cells, epithelioid cells and histiocytes of the granuloma type and were negative in those cells of the non-granuloma type. On the other hand, the small multinucleated giant cells which had 2-3 nuclei were the features of the morphological structure of the granuloma type. The small multinucleated giant cells were not seen in the non-granuloma type. CONCLUSION: In this paper, two pathological types (granuloma and non-granuloma type) in the MRS were proved further and it is found that the different structures of granuloma can be used to discriminate between MRS and sarcoidosis and tuberculosis. PMID- 12539343 TI - [The relationship between GPx activity in gingival fluid and clinical parameters of adult periodontitis]. AB - OBJECTIVE: To study the relationship of glutathione peroxidase (a natural FR scavenger) in gingival cervical fluid (GCF) of patients with periodontitis. METHODS: GCF was collected from 44 sites of 23 patients with adult periodontitis. The volume of GCF was measured with Periotron 8000. GCF-GPx activity was determined by DTNB methods before and after treatment, and the periodontal parameters GI, PD and AL were recorded. RESULTS: GCF-GPx activity was negatively correlated with PD and AL (P < 0.01, P < 0.005), but was not correlated with GI (P > 0.05). After treatment, clinical parameters decreased while GCF-GPx activity increased (P < 0.001), but the increment of GCF-GPx activity showed no correlation with the base level and the changes of clinical parameters. CONCLUSION: The imbalance between FR's production and scavenging might be an important pathological factor of periodontal diseases, and GCF-GPx might be a promising indicator of periodontal status. PMID- 12539344 TI - [TMJ radiographic bone alterations in patients of TMJDs]. AB - OBJECTIVE: The characteristics of temporomandibular joint (TMJ) bone changes showed by X-ray films in patients diagnosed as Temporomandibular Joint Disorders (TMJDs) were investigated. METHODS: X-ray films of 792 TMJDs patients who were diagnosised as TMJDs in our hospital from 1988 to 1995 were studied. Among them, 330 cases were male and 462 cases were female. Their ages ranged from 11 to 66 years old, averaged 27.4 years old. Bilateral TMJ, 1584 sides in all, were checked by X-ray. Among them, 210 cases (420 sides) were examined with transcraniall projective technique, 223 cases (446 sides) with tomogram, and 359 cases (718 sides) with both tomogram and transcranial projective techniques, Clinician observed and discussed the films together with the radiographer, and then the final radio-diagnosis was got. RESULTS: 1. 288 (25.31%) out of 1138 condyles examined with transcranial projective technique (temporal articular surface can not be observed accurately in this kind of films) and 82 (7.04%) out of 1164 condyles, 74 (6.36%) out of 1164 temporal articular surfaces checked with tomogram showed osseous changes. With analyzing on 718 sides of 356 cases checked with both methods, it was found that transcranial projective technique was more sensitive to TMJ osseous changes (P < 0.01). 2. The types of osseous changes: within the osseous changes of condyles showed in transcranial projection, 126 sides (43.75%) were mainly hyperplasia, and 140 sides showed absorptive changes. Moreover, both kinds of alterations appeared in 22 sides (7.64%). Among the condyles with osseous changes showed in tomogram, 28 sides were mainly hyperplasia and 54 sides showed absorptive changes, while two kinds of changes were not found in the same tomogram. But for temporal articular surface, 34 sides showed hyperplasia and 6 sides absorptive changes as their chief alterations. And the other 34 sides showed the two kinds of changes. Hyperplasia appeared more frequently in temporal articular surface than in condyles showed in tomogram (P < 0.01). 3. It was found that the osseous alterations were liable to occur in anterior condyle slope (88.19% in the films checked with transcranial projective technique, 81.71% in the films checked with tomography), the top (72.97% in the films checked with tomography) and the posterior slopes (71.62% in the films checked with tomography) of the articular eminence. CONCLUSION: Being considered the characteristics of the osseous changes, the abnormal mechanics around ICP and physiological or pathological remodeling principles of different parts of TMJ might be explained. Further researches on the selection of the two examinational methods of TMJ were suggested. PMID- 12539345 TI - [Study of accuracy of one-piece casted and laser welded bridges]. AB - OBJECTIVE: To assess whether laser welded bridge be fit for the clinical usage, this study compares the accuracy of laser welded multi-unit bridges with one piece casted bridges. METHODS: A precise stainless mold which was 20 mm length composed of two abutments (one premolar and one molar) and one pontic (two missed teeth) was made. 5 wax patterns of four-unit FPD frame-works in one-piece casting based on the mold were made. After that, they were invested with inner (Bellavest) and outer investment (phosphate and gypsum). Then they were casted with CW-PA (Ni-Cr-Nb) alloys. The other 5 wax patterns of four-unit FPD frameworks were invested and casted in two halves. Each couple of frameworks were ultra-sonic washed in distilled water and inserted to the standard metal FPD mold accurately. Laser welding machine (DL-2002) was used to weld each couple of frameworks at 305V, 20 ms. Both groups of specimens were tested on their mesiodistal span dimensions and the diameters of alloy crowns. Then they were inserted to their original stainless mold, where the marginal fitness was examined. RESULTS: 1. The one-piece casted fixed multi-unit bridge couldn't be inserted to abutments completely because of distortion, while the laser welded bridge could be inserted to abutments completely. The average marginal opening of laser welded bridges was about 74 microns and conformed to clinical standard of fitness. 2. Mesiodistal dimensions of one-piece casted bridges at the inner, bottom and top margins were 19.5600 +/- 0.3877 mm, 41.2206 +/- 0.4140 mm, 41.0598 +/- 0.3379 mm respectively; while those of laser welded bridges were 20.2950 +/- 0.1191 mm, 41.9625 +/- 0.0788 mm, 42.0584 +/- 0.1089 mm, which were obviously close to metal abutments (P < 0.05). 3. The alloy crowns' diameters of one-piece casted bridges were examined as following, the premolar: phi IJ = 9.8386 +/- 0.0567 mm, phi LK = 9.9596 +/- 0.0548 mm, the molar: phi EF = 11.7948 +/- 0.0252 mm, phi HG = 11.9412 +/- 0.0763 mm; The alloy crowns' diameters of laser bridges were as following, the premolar: phi IJ = 9.7927 +/- 0.0636 mm, phi LK = 9.9438 +/- 0.0726 mm, the molar: phi EF = 11.8287 +/- 0.0863 mm, phi HG = 11.8893 +/- 0.1049 mm. There was no significant difference between the two groups (P < 0.05). CONCLUSION: Laser welding greatly improve the fitness of multi-unit fixed bridges. PMID- 12539346 TI - [Stress-corrosion test of laser welded ceramic alloys]. AB - OBJECTIVE: The corrosion properties of dental material are very critical. This study is to investigate the corrosion properties of laser welded basemetalalloys. METHODS: The precise stainless rod molds had been made in size of phi 3 mm x 4 mm, with which 20 wax patterns were made. They were invested with inner investment (Bellavest) and outer investment (phosphate and gypsum). Then they were casted with CW-PA (Ni-Cr-Nb) alloys. All casted alloy rods were ultra-sonic washed in distilled water. Every two rods were contacted longitudinally by a special welding jig. Laser welding machine (DL-2002) was used to weld them at 305 V, 20 ms. The specimens were divided into 2 groups as following: 1. The group of stress-corrosion: 5 laser welded samples were subjected to the stress-corrosion test under 261.44 MPa in the 37 degrees C artificial saliva for 3 months in a special stress-corrosion test jig. After that, the ultimate tensile strength (UTS) was tested, the surface microcrack at the fusion zone (FZ), the heat affecting zone (HAZ) and the interface zone was observed with SEM (ARMY-1845, USA), the elements were also analyzed with EPMA (ARMY-1845, USA) at the fusion zone (FZ), the parent metal zone and the interface zone. 2. The other 5 laser welded samples were made as the controlled group. No special treatment were done to them, and the UTS was also tested, the surface of the micro crack and the elements at the FZ, the HAZ, parent metal zone and the interface zone were observed and analyzed with SEM, EPMA as well. RESULTS: The study showed us that the UTS of the stress-corrosion group was 502.48 +/- 12.49 MPa, while that of the controlled group was 446.05 +/- 82.50 MPa, there was no significant difference (P > 0.05). There was no microcrack on any sample's surface at the FZ, the HAZ and the interface zone. The main elements of the FZ were Ni(56.63%), Cr(19.36%), Nb (6.57%), Si(6.01%), Al(0.32%) and Mo(11.11%), while the elements of the parent metal zone were Ni(60.77%), Cr (20.59%), Nb(5.03%), Si(3.95%), Al(0.37%) and Mo(9.29%) respectively. CONCLUSION: The results suggest that the corrosion properties of laser welded alloys are excellent. PMID- 12539347 TI - [In vitro studies about the effects of implant materials on calmodulin, introcellular calcium of rat osteoblasts]. AB - OBJECTIVE: 1. To evaluate the effects of the four implant materials, hydroxyl poly calcium sodium phosphate (HPA), hydroxyapatite (HA), bioglass ceramics (BGC) and titanium (Ti) on the calmodulin (CaM) content of osteoblasts, the concentration of intracellular free calcium and the ATPases activity of cell membrane of rat osteoblasts cultured in vitro; 2. To reveal the biological compatibility of the four implant materials and the index to estimate the biological compatibility condition. METHODS: The powders of HPA, HA, BGC and Ti were sterilized and made into extracted solutions with 199 culture medium. The isolated osteoblasts, which were taken from three-day-old sparague Dawley rats' calvaria, were incubated in 199 culture medium also. Then the osteoblasts were mixed and incubated into the four kinds of extracted solutions of the implant materials. After three days of co-incubation, the cells were taken out, and the cellular calmodulin content were measured with radiobiological analysis. The concentration of intracellular free calcium were under fluorescent analysis, and the ATPases activities of cell membrane were estimated with colorimetric analysis. RESULTS: 1. Calmodulin content of the osteoblasts co-incubated with all the implant materials decreased, and moreover, only that of the Ti group decreased more obviously than that of the Ti and HPA group (P < 0.05). 2. The concentration of intracellular free calcium of all the co-incubated osteoblasts were as normal as that of the control group. 3. Ca(2+)-ATPases activities of osteoblasts co-incubated with HPA, HA, BGC increased (P < 0.05), and Na+/K(+) ATPases activities of the cells co-incubated with HPA, HA were strenghtened (P < 0.05). CONCLUSION: The indexes of cellular calmodulin content, concentration of intracellular free calcium and ATPases activity of cell membrane can reflect the biological compatibility of implant materials, and they may be used as indexes of estimating biological compatibility condition. PMID- 12539348 TI - [The analysis of the changes of tongue shape and position, hyoid position in Class II, division 1 malocclusion treated with functional appliances (FR-I)]. AB - OBJECTIVE: The objective of this study is to analyze the changes of tongue shape and position, hyoid position with FR-I therapy in class II, division 1 malocclusion. METHODS: 12 patients with class II, division 1 malocclusion treated with FR-I appliance(8 male, 4 female, average 11 years old) were chosen. The acceptable standard of this experiment was an horizontal growth pattern with normal maxilla, retracted mandible and short lower facial height. The average period of treatment was about 8 months. The assessment was made from a lateral cephalogram taken in habitual occlusion before and after treatment. The shapes and sizes of tongue can be measured from the occlusal films with template. These seven lines from six angles of 30 degrees which can be marked in millimeter. Fig 2 showed the reference points and lines: Is1 (the incisal margin of the lower incisors), Mc (the tip of the distobuccal cusp of the lower fist molar), V(the most caudal point on the shadow of the soft palate or its projection onto the reference line). Is1 connected Mc by a straight line extended to V to form the reference lines. After the lines were constructed, the middle point between Is1 and V was called O, and a perpendicular line was constructed from it to the palate contour. The baseline of the template was coincided with the constructed reference line, and the vertical line intersected the reference line at O. The template was placed over the constructed lines, and the exact measurements of the root, the dorsum and the tip of tongue were taken. This was done along the seven constructed lines, and the measurement gived a relative size of the tongue(e.g. the size related to the oral cavity). RESULTS: After treatment, the shape of tongue changed as a result of the increase of oral cavity. There was a tendency that the position of the tongue shifted forwarded and the tip of tongue lied further forward. At the same time, the position of hyoid moved forward in a horizontal dimension and downward in a vertical dimension. CONCLUSION: It confirms that the therapeutic mechanism of FR-I is to induce an anterior shift of mandible and lead to a forward position of tongue, hyoid and cervical soft tissue, which modify the saggital relationship of jaws. The shape of tongue changes due to the increase of oral cavity. PMID- 12539349 TI - [The changing trends of demography and oral diseases for Chinese dentistry]. AB - OBJECTIVE: The purpose of this paper is to discuss the changed trends of population, oral diseases and their effects on Chinese dentistry. METHODS: The demographic and epidemiologic data published in recent books and journals were reviewed and analyzed. RESULTS: As the 21st century approached, dentistry in China would face many changing trends and challenges. 1. a growing population and an aging population: The two sub-populations with the greatest need for prevention and treatment were children and old patients who were outpacing the supply of dental manpower. 2. The changed dental disease patterns: The prevalence of caries was increasing and more than two thirds of Chinese suffered from periodontal disease. The traditional dental approaches have not been able to satisfy the needs of Chinese. CONCLUSION: The challenge created by these demographic, economic and advances in dental technology is changing Chinese dentistry. PMID- 12539350 TI - [An investigation of malocclusion deformity of 2002 middle school students in Fuchuan Yao race autonomous county of Guangxi Zhuang race autonomous region]. AB - OBJECTIVE: To investigate the situation of maloccluson deformity which happens among the middle school students in Fuchuan Yao race autonomous county. METHODS: 2002 students (from 13 years old to 22 years old) were examined under natural light by using the way from Huang Jing fang, who edited a book called Oral Orthodontics. RESULTS: The rates of malocclusion deformity were 45.4% and 28.9% respectively, when the ideal normal occlusion and individual normal occlusion were treated as the criteria. CONCLUSION: Being compared with the relative reports, the results of the investigation show that the rate of malocclusion deformity is lower than that of the students in northern China. PMID- 12539351 TI - [Pathologic changes of lymphatic capillaries after injection of U14 cells in rat tongue perineoplastic area]. AB - OBJECTIVE: 1. To establish a model of neck lymphatic metastasis of cancer by injecting tumor cells into the rat tongue; 2. To observe the relationship between pathologic changes of perineoplastic lymphatic in different stages of implanted tumor growth and lymphatic metastasis of tumor cells. METHODS: 60 Kunming rats with injection of 5 x 10(6) U14 tumor cells into each rat tongue were chosen and randomized into 6 groups. In every 5 days, specimens of rat tongues with implanted tumor and neck lymph nodes with possible metastatic tumor cells were removed from each group of rats. Cryosections of rat tongue were processed with a modified enzyme-histochemical staining technique, using 5'-nucleotidase with HE to demonstrate lymphatic capillaries. Attention was paid to pathologic change of lymphatic capillaries in perineoplastic area of rat tongue in different growth stages of implanted tumor. Routing wax sections of lymph nodes processed with HE staining were examined under light microscopy to determine whether there were any metastasis tumor cells. RESULTS: 1. Metastasis to neck lymph nodes could be observed 15 days later after injection of U14 tumor cells into rat tongue. The situation of metastasis of lymph nodes varied greatly from tumor cells in bundle within marginal sinus to complete occupation of lymph nodes by U14 cells. 2. Tumor cells penetrated the lymphatic capillary by passing singly or in clumps between endothelial cells or through some destroyed endothelia. A great number of lymphatic with dilated cavity was found nearby the tumor tissue which facilitated tumor cells to enter the lymphatic cavity. 3. There were much more amount of lymphatic capillaries with dilated cavity around tumor mass comparing with that of normal tongue of the control group rats, and the number of lymphatic capillaries with opening cavity tend to increase with growth of implanted tumor. CONCLUSION: Direct and objective view of lymphatic capillaries can be obtained by means of the modified enzyme-histochemical staining technique, using 5' nucleotidase with HE to demonstrate lymphatic capillaries; A great number of lymphatic capillaries with dented cavity in normal physical state are forced to be dilated during process of tumor growth, thus in turn increase the possibility for tumor cells to penetrate the lymphatic capillary by passing singly or in clumps between endothelial cells. PMID- 12539352 TI - [An experimental study of transplantation of prefabricated axial flap with an vascular bundle]. AB - OBJECTIVE: To study the prognostic results of different prefabricated axial flaps with different operative methods. METHODS: Eighteen New Zealand rabbits and two operative patterns were utilized in this study. In group A, the vascular bundles of the left ear were transferred through tube flaps to the left neck. In group B, the central artery and veins of the right ear were transposed through skin tunnel to the right neck. After 4 weeks of operation, transplantation and vessel perfusion were made. RESULTS: Group A: The flaps of 10 cases were 100% survival, while there were 2 cases with 85% survival, and 2 flaps with necrosis. Group B: The flaps of 8 cases were 100% survival, while there were 5 sides with 50%-85% survival and 1 necrotic flap. The vessel perfusion demonstrated that there was no difference between the density values of the direct sprouting vessels from the transplanted vascular pedicles. There was no statistical difference between the two groups in flap survival and vessel perfusion. CONCLUSION: There is no obvious difference between the above two methods of prefabricated axial flap. PMID- 12539353 TI - [The regulation effects of growth factors on the procollagen gene expressions of human mandibular condylar cartilage cells]. AB - OBJECTIVE: At the early stage of osteoarthrosis (OA) and joint damage, the cartilage cells proliferation increases dramatically, but this repair process is finally replaced by the progressive destruction of the cartilage. The reason, in accordance to the latest research, is the abnormal phenotype of the involved cartilage cell. This result suggests that the regulation of cartilage cell phenotype may be an effective way in the treatment of OA and other cartilage destructive diseases. In this study, the effects of transforming growth factor beta(TGF-beta), insulin-like growth factor-I (IGF-I) and basic fibroblast growth factor (bFGF) on collagen expression of human mandibular condylar cartilage cells was investigated. METHODS: Chondrocytes were isolated from human fetus by enzymatic method. The second passage of the cells was used in this study. They were cultured in DMEM medium supplemented with 20% newborn calf serum (NCS). After the cells reached confluence, the medium was replaced by DMEM containing 0.4% NCS. Then the cells were exposed to different growth factors including IGF-I (10 ng/ml), TGF-beta (5 ng/ml) and bFGF (50 ng/ml). The steady state mRNA levels of different samples were examined by slot-blot hybridization. The results were analysed by Student t test. RESULTS: IGF-I had the least effects on the mRNA levels of three kinds of procollagen (type I, type II and type III). On the other hand, bFGF and TGF-beta could inhibit the expression of type II collagen by 0.352 and 0.685 times comparing with the control groups separately. TGF-beta increased type I collagen expression. Besides, bFGF and TGF-beta also increased the value of type I collagen/type II collagen. None of the three kinds of growth factors had obvious effects on the expression of type III collagen. CONCLUSION: IGF-I can maintain the chondrocyte differentiated phenotype, while TGF-beta and bFGF have inhibitory effects. PMID- 12539354 TI - [The effect of intra-articular injection of sodium hyaluronate and prednisolone on rabbits' temporomandibular joints]. AB - OBJECTIVE: To evaluate the different effects of sodium hyaluronate and prednisolone on the surface of rabbits' temporomandibular joints. METHODS: 20 juvenile Japanese large ear rabbits were divided into 5 groups, including one control group and the other 4 experimental groups. In these groups, half of their right inferior cavity of temporomandibular joints were injected with sodium hyaluronate(25 mg/ml, 0.1 ml), while others were injected with prednisolone(1%, 0.1 ml), then the rabbits were killed on the 1st, 3rd, 7th, and 14th days after injection. The specimens were evaluated by scanning electron microscope (SEM) to compare the different effects on the inferior superfacial structure of the articular disc and the surface of the condyle. RESULTS: Prednisolone could change the uniform distribution of the agglutinating substance on the inferior superfacial of the articular disc and the surface of the condyle. In the 1st-day and the 3rd-day groups, the agglutinating substance decreased in some areas and gathered in other areas. In the 7 th-day group, the distribution of the agglutinating substance on the inferior superfacial of the articular disc and the surface of the condyle were more regular than that of the 1st-day and the 3rd-day groups. In the 14th-day group, the distribution of the agglutinating substance were almost as the same as that of the control group. But sodium hyaluronate didn't change the uniform distribution of the agglutinating substance in every group. CONCLUSION: Prednisolone can change the uniform distribution of the agglutinating substance on the inferior superfacial of the articular disc and the surface of the condyle, while sodium hyaluronate don't have this effect. PMID- 12539355 TI - [A study of polysaccharide-protein complex and glycosyls in the tissue surrounding titanium implant of rabbit]. AB - OBJECTIVE: The absorption of the polysaccharide-protein complex to the implant surface may be the first reaction between the implant and the surrounding tissue. So we observed the distribution and the relative amount of the polysaccharide protein complex and glycosyls in order to find out the possible functions and changing rules of these substances. METHODS: 12 rabbits were selected according to the age, sex, weight, and health conditions. Bilateral mandibular premolar teeth of rabbits were extracted under abdomen anesthesia (2% glutababituratic acid), then 12 titanium implants were placed into the teeth extraction holes of one side and left the other side to be the control one. All of the implants were required to be placed 2 mm below the alveolar crest, and tissue closure was achieved with mattress and interrupted single sutures. Antibiotic injections were given to the rabbits in the following 3 days. The animals were sacrificed at the end of 1 week, 4 weeks and 24 weeks. Tissue blocks (4 mm x 4 mm x 4 mm) containing the titanium implants and the control ones were removed, then fixed with 10% formaldehyde for 2 weeks, and decalcified with 10% EDTA (pH7.0) for 8 weeks. Carefully removed the titanium implants, the tissue blocks were embedded into the paraffin wax, then the sections were obtained and stained using following methods. 1. Alcian Blue Method: Alcian blue (pH 2.5) staining displayed the acidic proteoglycans; Alcian blue (pH 1.0) displayed the proteoglycans containing sulfonyl; Alcian blue/periodix acid-shiff staining demonstrated the acidic and neutral proteoglycans respectively. Alcian blue-MgCl2 with different ionic concentration identified the different kinds of proteoglycans. 2. Lectin Method: 5 kinds of lectins were used to locate 5 kinds of glycosyls. 2.5 micrograms/ml ConA located the glucose; 10 micrograms/ml PNA located the galactose; 10 micrograms/ml WGA located the N-acetylglucosamine; 25 micrograms/ml PHA-E located N-acetylgalactosamine, 10 micrograms/ml WPL loacted the fucose. RESULTS: High level of proteoglycans and glycosyls were observed in the peri implant connective tissue during all phases of the experiment. It was also observed that polysaccharide-protein complex and glycosyls reached a high level in new bone matrix as well as osteoblasts. CONCLUSION: Polysaccharide-protein complex and glycosyls do play an important role in the implant-bone attachment, and titanium compatibility and may be closely related to the bone metabolic activity. PMID- 12539356 TI - [Changes of cytokines secreted by human oral mucosa keratinocytes from oral submucous fibrosis in vitro]. AB - OBJECTIVE: To assess the significance of oral mucosa keratinocytes (KCs) and cytokines in the development of oral submucous fibrosis (OSF). METHODS: KCs were obtained and cultured from the buccal mucosa of healthy subjects who did not chew areca nut and patients with OSF who chewed areca nut. Then radioimmunoassay and enzyme-linked immunosorbent assays were employed, and endothelin (ET) and transforming growth factor beta 1 (TGF beta 1) were quantified in culture supernatants. RESULTS: The mean levels of ET and TGF beta 1 secreted by OSF-KC were 96.983 +/- 17.802 pg/ml and 4661.641 +/- 783.893 pg/ml respectively, which were higher than these secreted by NM-KC (P < 0.05); There was a positive correlation between the levels of ET and TGF beta 1 secreted by OSF-KC. CONCLUSION: External stimuli such as areca nut components may induce the development of OSF by activating and stimulating the keratinocytes to secret a series of cytokines, including ET and TGF beta 1. It is speculated that ET and TGF beta 1 may be the contributing factors in the pathological features noted in OSF. PMID- 12539357 TI - [Animal experimental study on endocrine factors affecting alveolar repair]. AB - OBJECTIVE: To investigate the endocrine effects on activities of alkaline phosphatase (ALP) and acid phosphatase (ACP) and to obtain the mechanism of the resorption and regeneration of the alveolar bone. METHODS: 24 dog models with two side osseous infrabony pocket were divided into 4 groups randomly and were studied by histologic examination, enzyme cytochemistry and image analysis system. RESULTS: The activities of ALP and ACP, except ACP in the sixth month significantly decreased in the group of thyroidism comparing with that of the control group (P < 0.01), but only the activities of ALP significantly decreased in groups of ovariectomy and diabetes (P < 0.05, P < 0.01), and there was close relationship between enzyme activities and histopathologic variations. CONCLUSION: Alveolar bone repair was regulated by systemic endocrine factors and was associated with the activities of ALP and ACP osteoblasts of and osteoclasts. PMID- 12539358 TI - [Geometrical analysis of operative designs in unilateral cleft lip repair]. AB - OBJECTIVE: To study the differences in geometric principle in cleft lip repair between Millard and Tennison-Randall's method. METHODS: The moving patterns of the peaks of cupid's bow along the cleft on both sides varied according to the operative designs respectively. On the above basis, the larger vertical distance that the peaks of cupid's bow would drop in both operative designs respectively, which were calculated under the similar clinical conditions. RESULTS: It was found that the most obvious characteristic in Millard's design is to make the rotational axle on both cupid's bow peaks along the cleft drop farther away on the horizontal direction. However the Tennison-Randall's characteristic is to make the center of the rotational axle as near as possible to the cupid's bow peaks on the vertical direction. CONCLUSION: It is concluded that Millard's design conforms to the requirement of reconstructing the lip on the physiological aspect, but Tennison-Randall's design complies more on the geometric principles, which is the reason why both methods of cleft lip repair can not be replaced by each other. And this study will make both kinds of operation designs be applied clearly and easily. PMID- 12539360 TI - [Treatment of skeletal Class II malocclusion by orthodontic and surgical means]. AB - OBJECTIVE: Skeletal class II malocclusion is often occurred in Oriental and Black. It is difficult to gain the ideal treatment objective either using the orthognathic surgery or orthodontic treatment alone. However, the combined orthodontic and surgical treatment can reach it, so we investigate the methods and characteristics of combined orthodontic and surgical treatment in skeletal class II malocclusion. METHODS: We analyzed the recent 22 cases with skeletal class II malocclusion in Hospital of Stomatology, West China University of Medical Sciences with 7 male and 15 female from 20 to 30 years old. In the study, we discussed the time of extraction, the peculiarity of orthodontic treatment and the selection of surgery type. RESULTS: After the combined orthodontic and surgical treatment, all the patients regained satisfactory face appearance and function. The selection of surgery type is decided by the skeletal characteristics of malocclusion (the malformation degree and the position). The upper first premolars were extracted before the orthodontic treatment for the purpose of better curing of surface of the skeletal wound. And the pre and post surgical orthodontic treatment were very important and necessary to regain the ideal face appearance and function. CONCLUSION: The combined orthodontic and surgical treatment is an efficient way to cure skeletal class II malocclusion. Suitable orthognathic surgery and rational orthodontic treatment are important to treat adult skeletal class II malocclusion. PMID- 12539359 TI - [Clinical effects of intra-arterial nitrogen mustard (NH2) chemotherapy during operation for oral cancer]. AB - OBJECTIVE: To discuss the general and local side-effects and clinic appliance of the intra-arterial chemotherapy, and evaluate the short-time clinic effects of the intra-arterial chemotherapy. METHODS: From February, 1997 to December, 1997 11 patients with oral cancer (9 cases with lingual carcinoma, 2 cases with mouth carcinoma) were received one-shot chemotherapy during operation by NH2 catheterizing of lingual arteries (concentration of 1 mg/ml, dosage of 5 mg), and the other 16 oral cancer patients (13 cases with lingual carcinoma, 3 cases with mouth carcinoma) were only received operation as the control. At the same time, all patients were treated with Chinese flaps or Major pectoris myocutanous flaps homochronously. Then the general and local side-effects including the flap complexion were observed 1 week post-operatively and the follow-up was done in a shot time of 3-12 months. RESULTS: No obvious general and local side-effects (including blood, mal function of liver and kidney, flap complexion, vomit and hemiplegia) happened and all the 11 flaps (6 Chinese flaps, 5 major pectoris myocutanous flaps) were alive in the experimental group. The local recurrent rate of the experimental group was 9.1%, while the rate was 18.8% in the control group (following up of 3-12 months). There was no significant difference between the experiment and the control groups by Student t-test. CONCLUSION: The one-shot chemotherapy intra-operationally is a method with few side-effects, and it doesn't injure the homochronously reconstructed flaps. It might be an effective means of reducing the local recurrence of oral cancer. PMID- 12539361 TI - [Effects of pinyangmycin, dexamethasone and sodium morrhuate injection on treatment of cavernous hemangioma in maxillofacial regions]. AB - OBJECTIVE: To evaluate the effects of Pinyangmycin, Dexamethasonum and Sodium morrhuate injected homochronouly on treatment of cavernous hemangioma in maxillofacial regions. METHODS: The medical records of 115 patients with cavernous hemangioma in maxillofacial regions between September, 1996 and September, 1998 were reviewed. Fifty of the patients were men, and 65 were women, with a ratio of 1:1.3. The age of the patients was from 4 months old to 45 years old. 73 of the hemangioma bodies were sited in maxillofacial regions, 42 were sited in oral cavity. The sizes of the lesions varied from 1 cm x 1 cm to 5 cm x 8 cm. Treatment procedure: 1. Preparation: 8 mg Pinyangmycin was dissolved in 5 mg/ml Dexamethasonum and in 4 ml of 1% procain which were ready for use; 2. Injection: 1-5 ml (Pinyangmycin 1.6-8.0 mg) of mixed solution was first injected to the hemangioma, then followed by an injection of 0.5-2.0 ml Sodium morrhuate according to local appearance, size of the tumor and patient's age. Generally, the amount of each injection should not be over 8 mg Pinyangmycin. If the tumor did not subside after an injection, another injection might be repeated every 5-7 days, and this process might be repeated 3-5 times. The total dose of Pinyangmycin should not be over 40 mg. RESULTS: 115 patients were followed up for 6-24 months. 81 of 115 (70.43%) cases were recovered, and 30 (26.09%) cases were basically recovered. 4 (3.48%) cases got improvement. The recovery rate were 96.52%, and the total efficiency was 100%. CONCLUSION: This method may be a safe, simple and effective therapy to cavernous hemangioma in maxillofacial regions. PMID- 12539362 TI - [Clinical study of three-dimensional reconstruction of computed tomography in diagnosis of condylar fractures]. AB - OBJECTIVE: To explore a method of getting good three-dimensional (3D) reconstruction images of temporomandibular joint (TMJ) by computed tomography (CT), and evaluate diagnosis value of 3DCT in condylar fractures. METHODS: Of the 14 patients studied, 12 were male and 2 were female with age ranging from 4 to 37 years old. 4 were old fractures and the other 10 were fresh fractures. All patients' orthopanotomograms were taken first, and then were scanned with CT. The scanning conditions were 300-345 mAS and 120kV, and the scanning methods included cross-sectional (11/14), coronal-sectional (2/14) and spiral (1/14) (pitch: 1, thickness of layers: 2.5 mm) scanning. Scanning scopes: The cross-sectional and spiral scanning were from 1 cm above the Frankfort horizontal plane to the middle of the mandibular ramus or Menton; The coronal-sectional scanning was from the mastoid process to the anterior point of the mandibular ramus or Pogonion. The thickness of the scanning layers was 2.5 mm (12/14) or 5 mm (2/14). Finally, 3D reconstructed images were obtained by shaded surface display (SSD). The cross sectional images were obtained by being removed the cervical vertebra, the styloid process, the occipital bone and part of the mastoid process with Subtract Manual Irroi before reconstruction to avoid interference with the observation of TMJ and skull basis. RESULTS: 1. The best 3D CT reconstructed images were obtained by 2.5 mm-thin-layer cross-sectional scanning by being removed parts of the adjacent structures, and the bone lines were clear and smooth without adjacent bone structures shading TMJ. The quality of images taken spirally was similar to those taken cross-sectionally. While the coronal scanning neglected some important anatomic symbols which might be valuable to diagnose condylar displacement. 2. Of the 14 patients, 7 were unilateral condylar fractures and 7 were bilateral (21 sides altogether), among which 18 sides were high fractures of condyles and 3 were fractures of condylar neck. High oblique line fractures and comminuted fractures had major condylar rotation displacement which moved forward, downward and inside, whereas, vertical fractures only had minor displacement. Inward rotation displacement occurred in medial bent fractures. 10 of 14 patients (15 sides) had been conducted operation, and the same situations as seen from 3DCT reconstructed images were found. CONCLUSION: 3DCT images can display condylar fractures accurately and directly, and are very useful for surgeons to select treating methods of condylar fractures. PMID- 12539363 TI - [Effects of chincap therapy for early anterior crossbite]. AB - OBJECTIVE: To analysis the effects of chincap therapy for early anterior crossbite and mandibular prognathism. METHODS: 136 patients anterior crossbite and mandibular prognathism aged from 3 to 14 years old at the start of treatment were treated with active chincap with bilateral force of 700 to 1000 gram. Every patient wore chin cap 12 to 14 hours daily, and most of those patients used removable appliances to open the crossbite of incisors. According to the age of patients, they were divided into four groups, C1 (3-5 years old), C2 (6-8 years old), C3 (9-11 years old) and C4 (12-14 years old). When the treatment ended after two years, we analyzed the models by measuring the length of maxillary on the models before and after treatment (U1 and U2 that's the distance between the tip of 1|1 and the distal plane of 6|6 or V|V and the overjet before and after treatment (L1 and L2). Then we calculated the obtained distance for the mandible related to the maxillary position X(X = (L2-L1)-(U2-U1)) and analyzed treating time and mandibular retreating speed in the four groups. RESULTS: The obtained distance for the mandible related to the maxillary position in C1, C2, C3 and C4 groups were 3.27 +/- 0.99 mm, 4.62 +/- 1.33 mm, 4.43 +/- 1.28 mm and 2.63 +/- 0.92 mm respectively, and the treating time in four groups were 8.92 +/- 2.66 months, 10.47 +/- 2.21 months, 13.07 +/- 2.00 months, 20.50 +/- 3.96 months respectively. The retreating speed of the mandible related to the maxillary position in the four groups were 0.37 mm/month, 0.44 mm/month, 0.34 mm/month and 0.13 mm/month respectively. The effects of the treatment in C1, C2 and C3 groups (3-11 years old) were remarkable, especially in C1 group (3-5 years old). CONCLUSION: The effects of chincap therapy for early anterior crossbite and mandibular prognathism were remarkable, especially for the very early crossbite patients. PMID- 12539364 TI - [A study on the association of telomere length with the experimental carcinogenesis of oral cavity]. AB - OBJECTIVE: The life cycle of cell can be effected by the length of telomere. Loss of telomeric DNA during cell proliferation may play a role in chromosome instability, cellular apoptosis and cancer. The purpose of the study is to measure the dynamic changes of telomere length in experimental oral cavity cancer during golden hamster cheek pouch carcinogenesis induced by DMBA. METHODS: First, 52 golden hamsters were divided into 2 groups. Four of them were not done any treatment and were killed after 3 days. The others were covered with DMBA on the surface of cheek pouch on one side in order to induce carcinogenesis, the other side of cheek pouch was treated as the control. Then, the 48 golden hamsters were divided into 4 groups and were killed in 7, 10, 14, 20 weeks. Light microscope was used to observe the pathologic changes. Southern hybridization was used to analysis the dynamic development of the length of telomeric repeat. In different periods, the reduction rate of telomere length was calculated. RESULTS: 1. The cheek pouch mucosa uncovered with DMBA had no abnormal pathologic changes, while that covered with DMBA was observed pathologic changes of different degrees. 4 cases of hyperplasia lesions and 8 cases of dysplasia lesions were observed in the 7-week group, 7 cases of dysplasia lesions and 5 cases of mucosa carcinoma in situ in the 10-week group, 5 cases of mucosa carcinoma in situ, 4 cases of squamous cell carcinomas and 3 cases of death in the 14-week group, and 7 cases of squamous cell carcinomas and 5 cases of death in the 20-week group. 2. The telomere length of the normal mucosa was reduced with the hamsters age increasing, but the telomere length was significantly shorter than that of the normal control during the mucosa carcinogenesis of cheek pouch. The average shortened length was about 0.225 kb (the normal mucosa was 0.17 kb). 3. The highest reduction rate (0.3985) was in a later premalignant period. CONCLUSION: Age is one of the factors that can short telomere length. However, the speed that telomere length is shorted is faster than ever while telomere is abnormal. There is an obvious relationship between the abnormal degree of telomere length and the malignant degree of carcinoma. So the abnormal shorting of telomere length is an early molecular evidence of the oral carcinogenesis. PMID- 12539365 TI - [Identification of telomerase activity in the experimental carcinogenesis of oral cavity]. AB - OBJECTIVE: Telomerase is a ribonucleoprotein enzyme which synthesizes telomere DNA repeat sequences and maintain stably telomere length. The activity of telomerase may be necessary for the growth of immortalized cells overcoming cellular senescence. The researches have shown that telomerase activities are associated with most cancers. The purpose of the study is to detect the development of telomerase activity during golden hamster cheek pouch carcinogenesis induced by DMBA. METHODS: First, 52 golden hamsters were divided into 2 groups. Four of them were not done any special treatment and were killed after 3 days. The others were covered with DMBA on the surface of cheek pouch on one side in order to induce carcinogenesis, the other side of cheek pouch was treated as the control. Then, the 48 golden hamsters were divided into 4 groups and were killed in 7, 10, 14 and 20 weeks. The telomeric repeat amplification protocol (TRAP) based on PCR and ELISA was used to analysis the activity of telomerase. RESULTS: 1. The expression of telomerase activity existed in normal cheek pouch mucosa of golden hamsters, which meant that telomerase played an important role on controlling cell proliferation. 2. The level of telomerase activity gradually increased while hyperplasia and dysplasia was observed in the cheek pouch mucosa covered with DMBA. It reached its top at a later premalignant period and gradually decreased after that. 3. There was negative correlation between the degree the telomere length was shortened and the activity of telomerase (r = -0.9654), and there was positive correlation between the reduced rate of telomere length and the activity of telomerase (r = 0.9471). CONCLUSION: The activity of telomerase is one of important factors that can effect the stability of telomere length and plays a crucial role in the progression of oral cancer. So the activity of telomerase is an early molecular marker of carcinogenesis. PMID- 12539366 TI - [The predication equation of the permanent canine and premolar crown]. AB - OBJECTIVE: It is important to predict mesiodistal crown diameter of the permanent canines and premolars in the mixed dentition, which helps to determine malocclusion and its further development. But there is still no article about this equation of Chinese used to predict mesiodistal crown diameter of cuspids and premolars. This research is to measure mesiodistal crown diameters, to study relationship between lower incisors and canines and premolars, and to provide the most accurate and easy basis in orthodontic diagnosis and treatment for Chinese people. METHODS: 140 Chinese students (60 male, 80 female) who were from eighteen to twenty-five years old without orthodontic history Chengdu were randomly collected in. All teeth must be free of visible malform, aberration, and fully erupted. Impressions were taken, and measurements were made, parallel to the occlusal plane at the greatest mesiodistal width of each tooth on dental cast with electronic caliper (0.01 mm). Each tooth was measured twice and the average of the two readings was taken as the dimension for that individual tooth. Mesiodistal crown diameters were statistically analyzed with t-test and linear regression, using the SAS (Statistic Analyses System) computer package. RESULTS: It was found that mesiodistal crown diameters had significantly statistical difference in gender. Hence the mesiodistal crown diameters were assessed and investigated separately for the male and the female. It was also found that the total mesiodistal crown diameter of lower incisors was closely correlated with the total mesiodistal crown diameter of canines and premolars. The correlation coefficients between the independent variable and dependent variables were 0.599 for upper canines and premolars and 0.654 for lower canines and premolars in the males and 0.586 and 0.635, respectively, in the females. The P were found to be less than 0.001 for both arches. The correlation coefficients had significantly statistical difference. CONCLUSION: The equations predicting overall mesiodistal crown diameters of upper and lower unerupted canines and premolars for male and female Chinese people were established respectively: Maxillary (y = 10.87 + 0.51 x (male)), y = 11.88 + 0.46 x (female), Mandibular (y = 10.36 + 0.50 x (male), y = 10.03 + 0.49 x (female)), y (total mesiodistal crown diameter of canines and premolars), and x (total mesiodistal crown diameter of lower incisors). PMID- 12539367 TI - [Three-dimensional finite element stress analysis of supporting bone of mandibular posterior fixed bridge. Part III. Comparative analysis with stress of the cortical bone beneath different pontics of fixed bridge]. AB - OBJECTIVE: To analysis stress of the cortical bone beneath different pontics of mandibular posterior fixed bridge by using three-dimentional finite element method. METHODS: According to the measured data, the characters of all components of fixed bridge and the supporting tissues, and the principals of finite element, the three-dimentional finite element model was developed on the basis of an adult mandible. The model consisted of 1363 nodes and 1134 elements after the fixed bridge was placed. Three different pontics were designed. Young's mudulus and Poisson's ratio were obtained from the literature for each type of tissue or materials in the model. The surfaces of pontic next to the mucosa beneath the pontic consisted of nine elements and the contact area were respectively reduced 1/3. It was assumed all materials in the model were homogeneous, isotropic and linearly elastic. One vertical load of 20 kg and one horizontal load of 20 kg were applied respectively on the occlusal surface of the abutments and the pontics of three different fixed bridge. Three-dimentional finite element stress analysis was used to calculate the epsilon max and the epsilon min of the cortical bone beneath pontics of fixed bridge. RESULTS: Under the vertical loading, the cortical bone beneath the pontics appeared compressive stresses, the stress values of the cortical bone beneath the pontics were respectively -0.211( )-0.119 (epsilon max, kg/mm2), -0.355(-)-0.201 (epsilon min), -0.224(-)-0.125 (epsilon max), -0.375(-)-0.215 (epsilon min), -0.239(-)-0.133 (epsilon max), 0.395(-)-0.228 (epsilon min). Under the horizontal loading, the cortical bone beneath the pontics exhibited tensile stresses and compressive stresses. The stress values of the cortical bone beneath the pontics were respectively -0.004 0.283 (epsilon max, kg/mm2), -0.249(-)-0.009 (epsilon min), -0.0042-0.301 (epsilon max), -0.264(-)-0.0095 (epsilon min), -0.0045-0.322 (epsilon max), 0.281(-)-0.0101 (epsilon min). The stress in the cortical bone beneath the pontics increased when the area of contact was reduced, the stress values changed in 5%-7%, but the rules of stress distribution of three different pontics were similar. CONCLUSION: Stress values in the cortical bone beneath three pontics are small, and the change of stress in the cortical bone beneath the different pontics of mandibular posterior fixed bridge is small too. PMID- 12539368 TI - [Informatics of dental medicine]. AB - Dental informatics is a newly developed discinpline which is integrated by modern stomatology, computer sciences and informatics. In this paper, the concept, importance, basic principle and contents were discussed in details. The relationship among dental informatics and related sciences, the formation and history of dental informatics were also described in this paper. PMID- 12539369 TI - [Plasmid DNA expression in the muscle of rat tongue]. AB - OBJECTIVE: To develop rat tongue model for plasmid DNA expression in vivo. METHODS: The expression vectors which contained either human-muscle specific promoter or SV40 promoter directing the transcription of reporter gene (E. coli LacZ gene) were used. The straited muscle of the tongue was used as a model system, and expression plasmid DNA was injected directly into Wistar rat tongue. PCR technique, X-gal histochemical staining and Southern blot hybridization were used to analyze reporter gene expression in vivo. RESULTS: The expression of reporter gene was correlated with the amount of injected DNA and the time of incubation in tongue muscle. beta-galactosidase activity can be detected 24 hours after injection of X-gal staining and even be detected after two months, but the maximal expression level was observed one week later. Coinjection of two expression plasmids resulted in co-expression in the same striated myofibers. E. coli LacZ DNA sequence was found in rat tongue injected with the plasmid DNA by PCR, which demonstrated capacity of the rat tongue myofibers uptaking foreign DNA. The Southern blot analysis showed the injected plasmid DNA existed outside the genome of rat muscle cells. No integration was detected. CONCLUSION: Direct plasmid DNA injection into tongue muscle was a simple and efficient approach to transfer foreign gene into striated muscle cells, and tongue was a good model for analyzing exogenous gene expression. PMID- 12539370 TI - [Experimental study on apoptosis and Apo-1 expression of buccal carcinoma cell(BCC) induced by cisplatin]. AB - OBJECTIVE: To study apoptotic characteristics and expression of apoptosis molecule Apo-1 (Fas, CD95) on buccal carcinoma cell (BCC) induced by cytotoxic drug cisplatin and exploit the apoptotic mechanisms of oral cancer chemotherapy. METHODS: Human BCC line BcaCD885 was used as in vitro study object. Morphological features of apoptotic BCC induced by cisplatin were observed by use of lightmicroscopy and electronmicroscopy, meanwhile, apoptotic rates (AR) were analysed by the flow cytometry. RESULTS: 1. Condensation and crack of nucleus and apoptotic bodies appeared in apoptotic BCC; 2. There existed time and dosage dependent AR, and AR were respectively 5.4%, 7.4%, 9.4%, 20.3% and 22.6% after BCC had been acted by 15 micrograms/ml cisplatin for 2, 6, 12 and 24 hours, moreover, there was significantly difference (P < 0.01) among the 12-hour group, the 24-hour group and the control group; 3. Cell cycle of BCC was apparently arrested at G0/G1 by cisplatin (P < 0.05); 4. Apo-1 expression of BCC were enhanced 58% and 62% respectively after it was induced by 25 micrograms/ml cisplatin for 18 and 42 hours (P < 0.05). CONCLUSION: Apoptosis may be an important mechanism of BCC killed by cisplatin, and Apo-1 may play a role in the process of apoptosis induced by chemotherapy drugs. PMID- 12539371 TI - [Biological characters of adenoid cystic carcinoma cells transduced with HSV-tk gene]. AB - OBJECTIVE: To study the effects of herpes simplex virus thymidine kinase(HSV-tk) gene transduction on adenoid cystic carcinoma(ACC) cells. METHODS: Gene transduced cells were detected with growth rate, scanning electronic microscope, transmission electronic microscope, cell cycle analysis and the expression of major histocompatibility complex(MHC) class I or II molecules. RESULTS: The parameters detected in gene transduced cells and parental tumor cells were similar. CONCLUSION: The HSV-tk gene transduction would not change the biological characters of tumor cells greatly. PMID- 12539372 TI - [Telomerase hTRT mRNA expression in oral squamous cell carcinogenesis]. AB - OBJECTIVE: To investigate the relationship between telomerase hTRT gene expression and malignant development of oral mucosa epithelium. METHODS: 82 cases were detected by in situ hybridization technique for hTRT gene, including 7 cases of normal oral mucosa, 7 cases of hyperplasia lesions, 30 case sof oral precancerous lesions (dysplasia lesion), 8 cases of oral mucosa carcinoma in situ and 30 cases of oral squamous cell carcinomas. RESULTS: 1. Weaker expression of hTRT mRNA was observed in normal oral mucosa and hyperplasia lesions. Positive cells were present in basal layer and parabasal layer, and hTRT positivity was 21.4% (3 of 14). 2. The expression of hTRT mRNA in the precancerous lesions was gradually strong associated with phenotypic progression and the degree of dysplasia. Positive cell extended to multi-layered epithelium, and hTRT positivity was 46.7% (14 of 30). 3. Strong hTRT mRNA expression was observed in squamous cell carcinoma, and the positivity was 81.6% (31 of 38). CONCLUSION: Telomerase hTRT gene expression is closely related to the malignant degree of oral mucosa, the reactivated telomerase may play a crucial role in the progression of oral cancer. PMID- 12539373 TI - [Investigation on interaction between Streptococcus sanguis and Porphyromonas gingivalis in specific pathogen-free rats]. AB - OBJECTIVE: To examine whether endogenous S. sanguis could prevent, or reduce the colonization of the virulent P. gingivalis strain. METHODS: First, 10 specific pathogen-free Wistar rats were divided into 2 groups. Doxycycline was administered in the drinking water for 7 days. Successful implantation of the endogenous strain of S. sanguis, isolated from one of the rats before doxycycline administration, and P. gingivalis 381 within 14 days of observation were demonstrated in the rats of each group respectively. Then, 30 SPF rats were divided into 6 groups. Doxycycline was administered in the drinking water for 7 days to all the rats. Afterwards, the rats in group A and B were inoculated orally once a day for 5 days with P. gingivalis, the rats in group C and E were inoculated orally once a day for 5 days with S. sanguis. Then, the rats in group A were inoculated for 5 days with S. sanguis, and rats in group C and D were inoculated for 5 days with P. gingivalis. The rats in group F served as negative control. After inoculation, the levels of S. sanguis and P. gingivalis in the mouths of the rats were determined after 12, 24, 36 hours, 7 days and 14 days. RESULTS: Both pre-colonization of S. sanguis and superinfection with S. sanguis did reduce the level of P. gingivalis in experimental rats. However, the reduction only maintained quite short time, about 36 hours. It was not caused by the decreased level of S. sanguis after 36 hours because the level of S. sanguis kept stable during the observation period of 14 days. CONCLUSION: That S. sanguis function as the effector strain requires the successful implantation of S. sanguis as well as S. sanguis producing antagonistic action efficiently in vivo. PMID- 12539374 TI - [Separation and selection of salivary adhesion receptors of Streptococcus mutans to tooth surface]. AB - OBJECTIVE: To select and purify salivary receptors of Streptococcus mutans from experimental salivary acquired pellicle. METHODS: Experimental salivary acquired pellicle (SAP) was performed by coating hydroxyapatite (HA) with whole saliva. Then SAP was washed from HA by 1 mol/L NaCl and 0.5 mol/L phosphate buffer sequentially. The proteins were further separated by chromatography of Sephadex G75 and DEAE-Sephadex A25. Receptors of Streptococcus mutans were selected by bacterial adhesion test and competitive inhibition adhesion test. Identification was performed by PAGE, SDS-PAGE, IP-PAGE and detection of amylase activity and inmunodifusion test. RESULTS: IgA degraded fragments, a protein of 13 kD and amylases were the receptors of S. mutans. The first two only promote the adhesion but the amylases can both promote and inhibit S. mutans adhesion. CONCLUSION: The adhesion of S. mutans to tooth surface is a result of interaction between adhesins of S. mutans and multiple salivary receptors. PMID- 12539375 TI - [Significance of programmed cell death in C57BL/6N strain mouse palate process development and cleft palate formation]. AB - OBJECTIVE: To observe the changes of programmed cell death (PCD) in C57BL/6N strain mouse cleft palate formation and study related gene of PCD in future. METHODS: 16 mice (gestation of 10 days, GD10) were divided into the experimental and control groups randomly, then the mice of the experimental group were dosed with retinoic acid(RA) 80 mg/kg, and the tissue slices of the embryo mouse were prepared for TdT-mediated dUTP nick end labeling (TUNEL) staining at GD13(14)(the 14th hour of the 13th day gestation), GD13(22), GD14(8), GD14(14), GD14(22), GD15(8), GD15(22) and GD16(8) respectively. RESULTS: In the early development of palate process, the positive index of PCD was significantly higher in the experimental group than in the control group (P < 0.05). CONCLUSION: The effects of RA on mesenchymal cells of the early development of palate process in the experimental group induce PCD, which result in the formation of small palate process and failure of fusing of the shelves. PMID- 12539376 TI - [Effects of TGF beta on BMP-2 gene transfection cells in vitro]. AB - OBJECTIVE: To elucidate the mechanism of bone morphogenetic protein-2(BMP-2) and transforming growth factor beta(TGF beta) on cell differentiation. METHODS: A phagemid expression vector for human BMP-2 (pBK-B2) was constructed and transfected into the NIH3T3 cells by using lipofectamine. Positive cell clones were selected with G418. The stable transfection and expression of BMP-2 were determined by using in situ hybridization and immunohistochemistry methods. The proliferativity of the transfected cells treated with TGF beta were assayed by MTT method. Alkaline phosphatase (ALP) and osteocalcin (OC) production were also measured. RESULTS: The proliferation of the transfected cells was stimulated after being treated with TGF beta (50 ng/ml). But the ALP activity and OC production were inhibited. CONCLUSION: The effects of TGF beta are closely related to many factors, and BMP and TGF beta may be synergistic in cell proliferation and differentiation. PMID- 12539377 TI - [Investigation of aluminum oxide matrix of the machinable infiltrated ceramic(MIC)]. AB - OBJECTIVE: To explore the effects of different powder/liquid ratio during the procedure of aluminum oxide slip preparation on the properties and microstructure of the machinable infiltrated ceramic(MIC) aluminum matrice. METHODS: PI alpha Al2O3 powder was employed in this study. Aluminum slip with different powder/liquid ratios of 3.5, 4.5, 5.5, 6.5, 7.5, 8.5, 9.5 and 10.5 was prepared separately. Specimens were slip-casted and pre-fired, and whose physical properties were determined and morphology of specimens was examined under scanning electromicroscope. RESULTS: The packing density increased while the linear shrinkage decreased, when the powder/liquid ratio increased. Elevated pre firing temperature resulted in higher flexural strength of the matrice. The matrice demonstrated porous morphology under electromicroscope, and neck-fusion between Al2O3 particles was observed and increased with elevated prefiring temperature. CONCLUSION: MIC is a kind of easy-manipulated material with high packing density, optimum porosity and lower linear shrinkage, so it may be an ideal aluminum matrix for infiltrated ceramic. PMID- 12539378 TI - [New high strength titanium alloy Ti-Zr for dental use]. AB - OBJECTIVE: To develop a new titanium alloy which is suitable for prosthodontic use. METHODS: According to the mechanical properties of Co-Cr alloy and type IV dental gold alloy, a new titanium alloy Ti-Zr was developed. RESULTS: The datas of tensile strength(sigma b = 795 MPa, sigma 0.2 = 657 MPa, delta = 22%, E = 100 GPa) and hardness (249 HK) were collected. The microstructure showed a fine crystal grains. CONCLUSION: The new titanium alloy Ti-Zr is suitable for dental use. PMID- 12539379 TI - [Study on mechanical compression regulating the proliferation of rat mandibular condylar chondrocytes in vitro]. AB - OBJECTIVE: To study the proliferative changes of cultured rat mandibular condylar chondrocytes effected by mechanical compression. METHODS: Using flow cytometry(FCM) to explore the changes of cellular DNA content and cell cycles of passaged condylar chondrocytes. RESULTS: During certain force range (0-207 g/cm2), the proliferation of rat mandibular condylar chondrocytes rose, reaching the maximum after 6-hour application of force. The cellular proliferation went down when the chondrocytes were cultured under continuous pressure for 24 hours. CONCLUSION: The proliferation of rat mandibular condylar chondrocytes increase under certain force and certain period of time, but decreased if the applied pressure is beyond the magnitude. PMID- 12539380 TI - [Complete overdenture retained with implants and bar-clip attachments]. AB - OBJECTIVE: To sum up and evaluate the applied experience and applied effects of implants and bar-clip attachments in complete overdenture. METHODS: Bar-clip attachments were used as a super structure of implant to manufacture complete overdenture in three designs: mucosa, implant-mucosa, and implant supporting way. The retentive force of the overdenture was measured with measuring instrument of retentive force of prosthesis. RESULTS: All three designs could obtain good retentive effect, specially implant-mucosa supporting and implant supporting designs, which were of more better stability and supporting effect; 2-3 nelon clips could supply 10-15 N retentive force to the overdenture. CONCLUSION: The complete overdenture retained with implant and bar-clips attachments is a very effective way for restoration of endtulous patients, which should be applied more popularly. PMID- 12539381 TI - [A retrospective study of morphologic basis for the extraction decision in Class II, division 1 malocclusion]. AB - OBJECTIVE: To compare the dentofacial characteristics of persons with Class II, Division 1 malocclusion treated with either an extraction or a nonextraction approach. Such comparison might help identify which parameters influence the extraction decision. METHODS: Lateral cephalograms were available on 81 cases of Class II, Division 1 malocclusion, 42 were treated with 4 premolar extractions and 39 were treated with non-extraction. The cephalometric analysis of West China University of Medical Sciences and Model Analysis were used, RESULTS: Patients in the extraction group had larger FMA, FMIA and lower anterior facial height. In the group of extraction, the upper anterior teeth tipped more anteriorly, and the tooth size-arch length discrepancy and the depth of Spee curve were greater significantly. But the saggital discrepancy and soft tissue's profiles had no significant difference between two groups. CONCLUSION: The stability of the triangle of Tweed, the tooth size-arch discrepancy and the depth of Spee curve were the keys to decide weather extraction or not. PMID- 12539382 TI - [Morphological changes of external eyes before and after blepharoplasty]. AB - OBJECTIVE: To discuss the morphological changes of external eyes before and after blepharoplasty. METHODS: A survey of 92 patients treated by blepharoplasty was carried out to compare the external eye morphology of pre-operation with that of post-operation. RESULTS: It was found significant differences in the post operative patients, including the increased length and height of eyelids, and obvious changes of the length and angle of the exposed eyelashes. CONCLUSION: Blepharoplasty makes the eyelashes exposed and held up, and increases the up-to low distance of eyelids. These changes promote the beauty of eyes. PMID- 12539384 TI - [A follow-up study of anterior deciduous cross-bite]. AB - OBJECTIVE: To determine the relationship between anterior deciduous crossbite and anterior permanent crossbite, Angle classification of malocclusion and class III jaw relationship through a follow-up study in community children. METHODS: A follow-up study was done six years later in 102 children who were out of 120 children having anterior deciduous crossbite detected in a survey of six years before. Other 102 children with normal occlusion of deciduous and permanent teeth were treated as the control group. RESULTS: There were 51 out of 102 cases developed anterior permanent crossbite. In comparison with the group with 1-2 anterior deciduous crossbite, the group with more than two neighboring anterior deciduous crossbite had more significant probability to turn into anterior permanent crossbite, OR = 6.8 (95% CI: 2.7-17.6). The patients with more than 2 neighboring anterior deciduous teeth crossbite had significantly more chance of developing Angle III malocclusion and class III jaw relationship than those in the control group and those with 1-2 anterior deciduous teeth crossbite. CONCLUSION: Clinical orthodontists should pay more attention to the patients with more than 2 neighboring anterior deciduous crossbite. PMID- 12539383 TI - [Important adjacent relationship of external maxillary artery and clinic significances]. AB - OBJECTIVE: To investigate the important adjacent relationship of the external maxillary artery. METHODS: 60 pieces of complete head-neck samples were dissected layer by layer (only one side per sample), then the adjacent structures of the external maxillary artery, such as the submandibular gland, the anterior facial vein, the marginal mandibular branches of facial nerve and the submandibular lymphatic nodes were observed. RESULTS: 1. 60% of external maxillary artery lay on the bottom of submandibular gland, 38.3% went through the gland and 1.7% on the surface of the gland; 2. 90% of marginal mandibular branches of facial nerve went through the surface of external maxillary artery, 5% on the bottom and 5% encircled and gripped the artery; 3. 6.7% of anterior facial vein lay on the bottom of submandibular gland and accompanied external maxillary artery, 5% went through the gland and accompanied the artery, 88.3% went on the surface of the gland and didn't accompany the artery; 4. There were constant lymphatic nodes in front of and behind the external maxillary artery. CONCLUSION: The adjacent relationship of external maxillary artery is complex, and it is significant for doctors to know the above important structures around the artery. PMID- 12539385 TI - [Diagnosis and treatment of internal jugular phlebectasia (three cases report]. AB - OBJECTIVE: To discuss the diagnosis and treatment of patients with rare abnormal veins: the internal jugular phlebectasia. METHODS: Three patients with swollen during straining or crying were diagnosed as internal jugular vein dilatation by ultra-sonography, DSA and MRI. Then an operation of envelopment of the right internal jugular vein with the sternocleidomastoid muscle was performed in two cases. In the third case, the bilateral internal jugular ectasia, the right internal jugular vein were resected and the left internal jugular vein was enveloped with the resected vein. RESULTS: These three patients were treated satisfactorily by operation, and no recurrence in the following two years. CONCLUSION: To those who want their internal jugular phlebectasia to be treated, the operation of envelopment of internal jugular vein with sternocleidomastoid muscle is a good method. PMID- 12539386 TI - [Phenotypic changes of mandibular condylar cartilage cells during subculture]. AB - OBJECTIVE: To demonstrate the phenotypic changes of mandibular condylar cartilage(MCC) cells during subculture in vitro. METHODS: MCC cells were harvested from human fetus by digestion of collagenase and cultured in DMEM supplemented with 10% newborn calf serum, then they were passaged when reached confluence. Morphological changes were observed under phase-contrast microscope. Type II collagen, proteoglycan synthesis and type I, II and III procollagen mRNA levels were also studied in serial monolayer cultures. RESULTS: During subculture, type II collagen and proteoglycan synthesis were decreased, being accompanied by a lower type II collagen mRNA level. In contrast, type I and III procollagen mRNA levels were elevated progressively. The morphology of polygonal shaped chondrocytes was lost to the bipolar fibroblastic cells. CONCLUSION: Passage and culture can affect the phenotypic property of MCC cells. PMID- 12539387 TI - [Effects of growth factors on DNA and collagen synthesis of human mandibular condylar cartilage cells]. AB - OBJECTIVE: To investigate the effects of transforming growth factor-beta 1 (TGF beta 1), insulin-like growth factor-I (IGF-I) and basic fibroblast growth factor(bFGF) on DNA and collagen synthesis of mandibular condylar cartilage(MCC) of human fetus. METHODS: Cell culture, 3H-TdR and 3H-Proline incorporation methods were used. MCC cells were harvested from 4 to 5 months old human fetus. Cells were seeded at 2 x 10(4)/well on 96-well Plate. After synchronization, medium was replaced by DMEM containing 0.4% NCS with various growth factors and concentrations. RESULTS: bFGF stimulated the DNA synthesis significantly, and IGF I had less effect, while the effect of TGF-beta 1 was insignificant. For collagen synthesis, bFGF caused a dose-dependent increase(60%). A greater effect(98%) was achieved when IGF-1 was added. In contrast, TGF-beta 1 could inhibit collagen synthesis (24%). CONCLUSION: Growth factors play an important part in the proliferation and matrix synthesis of MCC cells, which might be of potential application in treating cartilage destructive lesions. PMID- 12539388 TI - [Construction of Streptococcus mutans surface protein antigen eukaryotic expression vector pcDNA3-PAc. I. Extraction and purification of plasmid DNA pPC41 and pcDNA3]. AB - OBJECTIVE: To Extract and purify plasmid DNA pPC41 and pcDNA3. METHODS: Bacteria were collected and lyzed by the alkaline lysis method. The plasmid pPC41 carrying structural gene pac and clone expression vector pcDNA3 were extracted and purified from E. coli clone by the polyethylene glycol precipitating method, the electroelution into dialysis bags method, the low melting-temperature agarose gel and glass fiber column chromatography. The concentration, purity and molecular weight were determined by spectrophotometry, restriction enzyme and agarose gel electrophoresis. RESULTS: The concentration of plasmid DNA of this experiment in general was 0.12-0.24 g/L, and A260/A280 of the polyethylene glycol precipitating method, the electroelution into dialysis bags method, the low metling-temperature agrose gel and glass fiber column chromatography were respectively 1.9, 2.2, 2.2 and 2.6. Molecular weight of pPC41 and pcDNA3 was respectively 10.6 kb and 5.4 kb. CONCLUSION: The four methods can obtain pure plasmid DNA. Purity of plasmid DNA obtained by glass fiber column chromatography method is the highest in the four methods. Glass fiber column chromatography is one of the effective methods getting highly purified plasmid DNA. PMID- 12539389 TI - [Construction of Streptococcus mutans surface protein antigen eukaryotic expression vector pcDNA3-PAc. II. Testified efficiency of repeated using glass fiber chromatographic column]. AB - OBJECTIVE: To explore one method getting highly purified plasmid DNA and costing low. METHODS: According to principle of extraction and purification plasmid DNA of glass fiber chromatographic column, its operation sequence was improved so that disposable glass fiber chromatographic column could be used repeatedly. The concentration and purity of plasmid DNA obtained by 12 times repeated using glass fiber chromatographic column were determined by spectrophotometry. Moreover, Those plasmid DNA obtained were tested by restriction enzyme and agarose gel electrophoresis, Ligation in vitro and transformation test. RESULTS: 12 times repeated use of glass fiber chromatographic column made no significant difference to the concentration and purity of obtained plasmid DNA (P < 0.05), and plasmid DNA was completely digested by restriction enzyme. The digested fragments were well ligated together by the action of T4 DNA Ligase in ATP ligating buffer in vitro and ligated DNA was well transformed into E. coli. CONCLUSION: Repeated use glass fiber chromatographic column can obtain a lot of highly purified plasmid DNA and cost is low. Digestion, ligation and transformation of those plasmid DNA are not affected. The results may be significant in molecular cloning and study of gene vaccination. PMID- 12539390 TI - [A primary study of relationship between proteolytic enzymes in dental plaque and deciduous caries]. AB - OBJECTIVE: The purpose of this paper was to study the role of proteolytic enzymes in dental plaque in deciduous caries occurrence. METHODS: Fluorometry and spectrometry were used separately to determine the activities of leucine amino peptidase, dipeptidyl peptidase IV and trypsin-like proteinase in caries-positive and caries-free children. RESULTS: The activities of the three enzymes in dental plaque in different children were same (P > 0.05). CONCLUSION: The three enzymes may be not related to deciduous caries. PMID- 12539391 TI - [The study of the activities of salivary proteolytic enzymes in children]. AB - OBJECTIVE: The activities of proteolytic enzymes in saliva were assayed to determine whether they were related to caries sensitivity and gender in the children. METHODS: Fluorometric and photometric methods were applied. Three enzymes included leucine amino peptidase, dipeptidyl peptidase IV and trypsin like proteinase. RESULTS: 1. Only dipeptidyl peptidase IV and trypsin-like proteinase were demonstrated in saliva, they were obviously lower than those in dental plaque (P < 0.05); 2. The activities of proteolytic enzymes of male and female children as well as caries-active and caries-free children showed no difference. CONCLUSION: Proteolytic enzymes play little role in caries formation of deciduous tooth. PMID- 12539392 TI - [Three-dimensional finite element analysis of stress in supporting bone of mandibular posterior fixed bridge. I. Stress analysis of the surface of alveolar bone around bridge abutments]. AB - OBJECTIVE: Stress levels of the surface of alveolar bone around bridge abutment of posterior fixed bridge were studied. METHODS: The three-dimensional finite element model was developed on the basis of a try adult mandible. 6 vertical and 1 horizontal loadings were applied respectively, and the Maximum stress(epsilon max) and the Minimum stress(epsilon min) of the surface of alveolar bone around bridge abutment were calculated. RESULTS: The stress concentration exhibited at the apical and cervical regions. The stress of the bicuspid was about 2.1 times as high as that of the molar. When transverse loading was applied, the stress level was about 3.5 times as high as that when vertical loading was applied. CONCLUSION: The results were greatly valuable to design of posterior fixed bridge. PMID- 12539393 TI - [Three-dimensional finite element analysis of stress in supporting bone of mandibular posterior fixed bridge. II. Stress distribution of the cortical bone beneath pontic of fixed bridge]. AB - OBJECTIVE: Stress distribution of the cortical bone beneath the pontic of the fixed bridge was observed. METHODS: The three-dimensional finite element model was developed. 6 loadings were applied respectively and the Maximum Stress (epsilon max) and the Minimum Stress(epsilon min) of the cortical bone beneath the pontic of the fixed bridge were calculated. RESULTS: The stress values were very small before the bridge was inserted. After the bridge was inserted the stress increased, but the stress level in the cortical bone was much lower than that in the alveolar bone. The stress values of the cortical bone were only 9.5% (vertical loading) and 2.2% (horizontal loading) of the alveolar bone's. CONCLUSION: The stress value of the cortical bone beneath the pontic was very small. The load was born mainly by supporting tissue around the bridge abutment. PMID- 12539394 TI - [Recombinant Salmonella typhimurium anticaries vaccine. II. Construction and confirmation of expressing plasmid carrying a region of S. mutans pac gene]. AB - OBJECTIVE: The objective of this study is to construct prokaryotic expressing plasmids carrying A region of S. mutans pac gene. METHODS: The A region of pac gene was amplified by using PCR from plasmid pPC41 containing S. mutans surface protein pac gene. The new expression plasmid was constructed by using A region of pac gene and expressing plasmids pET17-b and directed cloning technique. The recombinant plasmid was identified by agarose gel electrophoresis after it was cut by using BamHI and EcoRV. RESULTS: The expressing plasmid carrying pac gene A region was acquired. CONCLUSION: The recombinant plasmid provides useful experimental materials for further research work. PMID- 12539395 TI - [An immunohistochemical study of distribution of insulin in lateral pterygoid muscle after functional mandibular protrusion in rats]. AB - OBJECTIVE: The aim of this study is to investigate the effects of insulin on the lateral pterygoid muscle of growing rats after the functional mandibular protrusion. METHODS: An immunohistochemical (the ABC method) method was used to detect the location and distribution of insulin in the lateral pterygoid muscle of rats. RESULTS: 1. Insulin located in the cell membrane of the lateral pterygoid muscle. 2. The distribution of insulin in the lateral pterygoid muscle of rats was enhanced after the functional mandibular protrusion. CONCLUSION: Insulin may play an important role in adaptive remodeling of the lateral pterygoid muscle after the treatment of functional mandibular protrusion. PMID- 12539396 TI - [Cell proliferation in odontogenic jaw cyst epithelium]. AB - OBJECTIVE: The aim of the study is to investigate the epithelial cell proliferation in the lining of odontogenic cysts with Ki-67 monoclonal antibody. METHODS: Ki-67 expression was studied in 30 odontogenic cysts (odontogenic keratocysts, n = 10; dentigerous cyst, n = 10; radicular cysts, n = 10), and normal oral epithelium (n = 10) by using a biotin streptavidin method on routinely processed paraffin sections after microwave treatment. Ki-67+ cells were counted manually and related to the area of epithelial lining as determined by computer image analyzer. The result was analyzed by using a computer image analyzer to calculate positive cells per square millimeter of the epithelial lining of odontogenic lesions. Data were analyzed by using student test. RESULTS: The epithelial lining of odontogenic keratocysts contained the highest number of Ki-67+ cells, most of which were located in the suprabasal layers. The mean value of Ki-67+ cells in odontogenic keratocysts linings (1812.29 +/- 606.47 cells/mm2) were significantly higher than that of dentigerous (944.43 +/- 58.98 cells/mm2) and radicular cysts (610 +/- 26.4 cells/mm2) (P < 0.01). Negative reactions were obtained in the normal oral epithelium. CONCLUSION: The results demonstrate that there are differences in Ki-67 expression between epithelial linings of the major odontogenic cysts, which were consistent with odontogenic keratocysts having a greater proliferative capability than dentigerous and radicular cysts. It also indicates differences in proliferation and differentiation processes within these lesions. PMID- 12539397 TI - [Effects of insulin and transforming growth factor-beta on alkaline phosphatase activity and total protein content in human periodontal ligament cells]. AB - OBJECTIVE: The aim of this study is to evaluate the biological effects of insulin and/or transforming growth factor (TGF-beta) on alkaline phosphatase (ALPase) activity and protein synthesis of periodontal ligament (PDL) cells in vitro. METHODS: The fifth passage of PDL cells were cultured in a 5% humid CO2 environment at 37 degrees C by using Dulbecco minimum essential medium (DMEM) supplemented with 10% fetal bovine serum (FBS) for 24 hours, and then insulin, TGF-beta, and insulin and TGF-beta were added respectively in a medium of 1% FBS. After 5 days, their ALPase activities were measured by using a kinetic method, and the total protein content was measured by using Coomassie brilliant blue staining. RESULTS: Both insulin and TGF-beta significantly increased the ALPase activity and the total protein content of PDL cells at the concentrations from 1.0 to 100 U/L, and of 0.1-100 micrograms/L. The most optimal concentrations of insulin and TGF-beta were respectively 10 U/L and 1 microgram/L. The synergistic effect of insulin (10 U/L) and TGF-beta (1 microgram/L) was observed (P < 0.01). CONCLUSION: The results suggest that insulin and/or TGF-beta may promote the differentiation and protein synthesis of PDL cells. The combination of them may be valuable in the regeneration of periodontal tissue. PMID- 12539398 TI - [Immediate microcirculation reaction of periodontal ligaments to experimental intrusion of dog teeth]. AB - OBJECTIVE: The purpose of this study is to investigate the immediate microcirculation reaction of periodontal ligaments to experimental intrusion of dog teeth. METHODS: The forth premolar was chosen as the anchor tooth to intrude the second premolar using an intrusion circle loop. All tissue sections were observed and analyzed using the ink perfusion technique and an image analysis apparatus. RESULTS: The microvascular pattern of periodontal ligaments was changed after the maxillary second premolar had been intruded for 2 hours with 100 g of force. The vascular diameter and volume density diminished, but the vascular number and some parts of the microvasculature, especially capillaries were compressed due to the blood escaping from the periodontal ligament. It showed that the initial effect of 100 g of intrusive force caused adaptable changes in the microcirculation of periodontal ligaments, but no significant impediment of blood flow occurred. CONCLUSION: During the tooth intrusion with 100 g of force, a suitable microenvironment was established for the benefit of the survival of periodontal ligaments, which could avoid overloads imposed on the periodontal ligament and decreased the injury of periodontal tissue. It elucidates that the microcirculation of periodontal ligaments plays an important role in the protection of the periodontal tissue, in terms of biological function. PMID- 12539399 TI - [Effects of bite force loss on the expression of IL-6 mRNA in rat periodontium]. AB - OBJECTIVE: The aim of this study is to evaluate the expression of IL-6 mRNA in rat periodontal cells under normal bite force and without bite force, and to explain periodontal remodeling from the molecular level. METHODS: Animal models with normal bite force and without bite force were established by using Wistar rats. In situ hybridization technique was applied to detect changes of IL-6 mRNA expression in rat periodontal tissue. RESULTS: Without bite force, the expression level of IL-6 mRNA increased significantly in periodontal cells. CONCLUSION: This study initially investigates the molecular mechanism of periodontal remodeling, and suggests that the histological morphology may be closely related to the mechanical stimulation. PMID- 12539400 TI - [Effects of mechanical stretch on collagen type I and total protein synthesis of human periodontal ligament fibroblasts]. AB - OBJECTIVE: The aim of this study is to investigate the effects of mechanical stretch on type I collagen and total protein synthesis of human periodontal ligament fibroblasts (PDLFs). METHODS: Enzyme link immunosorbent assay (ELISA) and Coomassie brilliant blue stain method were used to examine the anabolism of PDLFs growing on the flexible culture plates subjected to mechanical stretch. The five loading devices of PDLFs were designed and the stain varied from 1% to 8%. RESULTS: Mechanical stretch had no effect on type I collagen synthesis. However, in the range of 1% to 8% of stretch, the protein synthesis was reduced. The mechanical stretching of 2% strain affected the protein synthesis most obviously. CONCLUSION: Mechanical stretch can modify the composition of extracellular matrix of PDLFs. PMID- 12539401 TI - [A possible criteria for diagnosis of early caries in pits and fissures]. AB - OBJECTIVE: The aim of this study was to examine pits and fissures of occlusal surfaces with severe stains by both histological and clinical methods, and to evaluate the criteria for diagnosis of early caries in pits and fissures. METHODS: Thirty-one fresh extracted molars and premolars, severely stained but no detectable defects in the surfaces, walls and floors of occlusal fissures, were collected. The teeth were longitudinally sliced across the stained fissures and ground into sections of 100-140 microns. The sections were observed using microscopy and microradiography. RESULTS: All samples showed early carious lesions in pits and fissures under microscope. Demineralization in walls or floors was identified by microradiography in 30 samples. CONCLUSION: Although without detectable defects, severely stained pits and fissures showed demineralization by using microscopy and microradiography. Thus, early caries in pits and fissures may be identified by the severely stained appearance. PMID- 12539402 TI - [Investigation of the tongue squamous cell carcinoma prognostic factors]. AB - OBJECTIVE: The aim of this study was to investigate the factors which influence the prognosis of the tongue squamous cell carcinoma (TSCC). METHODS: The Cox's proportional hazard model was used to analyze 121 cases of TSCC treated by the affiliated dental hospital of West China University of Medical Sciences in the period from 1990 to 1994. Thirteen parameters (age, sex, occupation, primary tumor site, clinical stage, tumor differentiation, clinical therapy, cervical lymph node metastasis, surgical margin status of the primary tumor, amount of blood transfusion, smoking, drinking, reconstruction during operation) were evaluated. RESULTS: The monofactor analysis showed that factors influencing the postoperative survival of TSCC included primary tumor site, the surgical margin status of the primary tumor and the cervical lymph node metastasis (P < 0.05). Multifactor analysis demonstrated that two of these thirteen relevant factors influenced the postoperative survival of TSCC independently. They are successively the surgical margin status of the primary tumor and the cervical lymph node metastasis (P < 0.05). CONCLUSION: This study suggests that it is critical for surgeons to resect the primary tumor and the metastatic cervical lymph nodes thoroughly to improve the survival rate of TSCC patients. PMID- 12539403 TI - [Clinical study on effects of root canal instrumentation on the root canal microflora]. AB - OBJECTIVE: To investigate the effects of instrumentation and ultrasonic treatment on the root canal flora. METHODS: A total of 21 single-rooted teeth with chronic apical periodontitis were selected and prepared using step-back/ultrasonic technique, Samples were taken before and after the step-back preparation, and after the ultrasonic treatment. RESULTS: The number and species of bacteria in the root canal were both reduced dramatically and there were significant differences among the three groups (P < 0.05). CONCLUSION: The instrumentation and ultrasonic treatment of root canal can effectively reduce the number and species of root canal microflora. PMID- 12539404 TI - [Guided tissue regeneration with Biomesh membrane for Class II furcation defects]. AB - OBJECTIVE: The objective of this study was to investigate clinical significance of the guided tissue regeneration (GTR) by using Biomesh membrane to the treatment of Class II furcation defects. METHODS: Totally 8 molars with Class II furcation defects of 6 periodontitis patients were treated. The clinical effects of GTR were evaluated by measuring changes of pocket probing depths (PPD), clinical attachment levels (CAL), and alveolar bone on X-ray films prior to and 6 months after the surgery. RESULTS: All patients had no swollen or pain reaction on these surgery areas after the surgery, and the PPD was reduced and the CAL was gain significantly 6 months after the surgery. The new alveolar bone formation was demonstrated. CONCLUSION: The GTR with Biomesh membrane is a good therapy method for Class II furcation defects. PMID- 12539405 TI - [Relationship between the symptom of xerostomia and non-stimulated salivary flow rates in patients with burning mouth syndrome]. AB - OBJECTIVE: Many patients with burning mouth syndrome (BMS) complain of xerostomia, but the consistent relationship between the symptom of xerostomia and non-stimulated salivary flow rates remains unclear. This study tried to investigate the variety of non-stimulated salivary flow rates in patients with BMS. METHODS: The non-stimulated salivary flow rates of 52 BMS patients and 37 healthy controls were recorded. RESULTS: The non-stimulated salivary flow rates were low in the group of BMS patients, but there is no statistical difference when compared with that of the normal individuals. There was also no significant difference in non-stimulated salivary flow rates among theses three subtypes of BMS patients. Further, no significant difference of non-stimulated salivary flow rates was observed in patients with and without xerostomia. CONCLUSION: The non stimulated salivary flow rate may be not associated with the symptom of xerostomia in patients with BMS. PMID- 12539406 TI - [Clinical application of Targis veneers]. AB - OBJECTIVE: The aim of this study was to evaluate the Targis material in the form of veneers. METHODS: Nineteen patients treated with 91 Targis veneers were involved in this study, and the clinical evaluation on veneers was conducted at the time of recall. The evaluation parameters included marginal adaptation, marginal discoloration, surface texture and color match. The periodontal status of all teeth with veneers was evaluated by gingival bleeding index and plaque index before and after the treatment. RESULTS: The mean observation period for the veneers was 14.6 months. Four veneers failed due to fracture, three because of debond, one because of discoloration. The cumulative rate of success was 91.2%. There was no significant difference in gingival bleeding index before and after treatment, but the plaque index significantly decreased after the treatment. CONCLUSION: The clinical results of Targis are satisfactory. PMID- 12539407 TI - [Factors involved in intelligent development of children with cleft lip and palate]. AB - OBJECTIVE: The objective of this study was to investigate the possible factors involved in intelligent development of children with cleft Lip and palate. METHODS: The intelligence quotient (IQ) and developmental quotient (DQ) of 152 children with cleft lip and palate under 14 years old were conducted, and the control group was made up of 80 healthy children. The considered factors included degrees of deformation, situation of perinatal period, way of feeding, psychological conditions of the children, general situation of their parents and education of these children. The data were statistically analyzed and compared. RESULTS: The IQ value of these children generally was normal, but the mean value was still lower than that of the control group (P < 0.01). The mean IQ value of the group of children with only cleft lip or cleft palate was higher than that of the group with both cleft lip and cleft palate, and the mean IQ value of these children with II degree deformity was higher than that of children with III degree deformity. The mean IQ value of these children feed with mother's milk was higher than that of those feed with artificial milk. These children more than 5 years old were disturbed psychologically, and their parents were also with pressure. CONCLUSION: The intelligent development of children with cleft lip and palate is affected by many factors, including types and degrees of deformity, feeding ways, illnesses and social psychological factors in different periods of development. Therefore, it is very important to take steps before 5 years old, including correctly feeding, repairing the deformity as early as possible, preventing the illnesses, necessary psychological treatment, improving social environment. PMID- 12539408 TI - [Effects of the mold temperature on the castability of titanium zirconium alloy for dental clinical use]. AB - OBJECTIVE: The aims of this study were to investigate the influence of the mold temperature on the castability of Ti-Zr alloy and find out the reasonable method for casting Ti-Zr alloy in clinic. METHODS: Ti-Zr alloy, pure titanium and Ti-6Al 4V alloy were casted respectively, by using a mold made of a zircon (ZrO2.SiO2) investment and a casting machine (China) designed, with a program of vacuum, pressure and centrifuge casting at three mold temperatures (room temperature, 300 degrees C, 600 degrees C). The castability value was obtained by counting the complete segment of the cast alloy grid. The wax pattern provided a grid with 112 segments. The percentage of complete cast segments was calculated by divided by 112, and multiplied by 100. The mesh castability values of Ti-Zr alloy, pure titanium and Ti-6Al-4V alloy were examined. RESULTS: The castability of three titanium alloys was high when the mold temperature was high. The castability values of Ti-Zr alloy at three mold temperatures were 88.57% for the room temperature, 94.55% for 300 degrees C, and 96.67% for 600 degrees C. Ti-Zr alloy casted at the room temperature showed significantly lower castability value than that casted at 300 degrees C and 600 degrees C (P < 0.05), but no significant difference was found between the castability values of 300 degrees C and 600 degrees C (P > 0.05). The castability values of pure titanium at three mold temperatures were 95.33% for room temperature, 98.70% for 300 degrees C, and 98.90% for 600 degrees C. The castability values of Ti-6Al-4V alloy at three mold temperatures were 95.37% for room temperature, 96.00% for 300 degrees C, and 98.78% for 600 degrees C. When casted at room temperature, the castability values of Ti-Zr alloy were significantly decreased comparing with those of pure titanium and Ti-6Al-4V alloy (P < 0.05), but no significant difference was found when cast alloys were casted at 300 degrees C and 600 degrees C (P > 0.05). CONCLUSION: The mold temperature is one of the major factors influencing cast quality. This study suggests that in order to obtain high castability of dental Ti-Zr alloy castings in clinic, the optimal mold temperature should be higher than 300 degrees C. PMID- 12539409 TI - [Transverse deflection study of casted titanium zirconium alloy for dental clinic use]. AB - OBJECTIVE: The aim of this study was to obtain the reasonable thickness of Ti-Zr alloy used for denture base material. METHODS: According to the "Revised ADA Specification No. 12 for denture base polymers", the transverse deflection tests of Ti-Zr alloy after casting were performed. Pure titanium, Ti-6Al-4V alloy and Co-Cr alloy were examined as the control. Three groups of wax patterns with different thickness were designed. Ti-Zr alloy, pure titanium and Ti-6Al-4V alloy were casted into molds at 300 degrees C, which were made of zircon (ZrO2.SiO2) investment, using a casting machine (China), with a program of vacuum, pressure and centrifuge casting. Co-Cr alloy was casted by normal method in clinic. The transverse deflections of these four alloys castings with three different thicknesses were measured using a transverse testing machine. RESULTS: When the thickness is 0.65 mm for Ti-Zr alloy, pure titanium and Ti-6Al-4V alloy, and 0.57 mm for Co-Cr alloy, the transverse deflections could satisfy the requirement. CONCLUSION: The transverse deflection is one of the key indexes for evaluating the ability of dental material to bear biting forces. The reasonable thickness of Ti-Zr alloy used as denture base material is 0.65 mm. PMID- 12539410 TI - [An experimental study on effects of pingyangmycin on vessels]. AB - OBJECTIVE: The aim of this study was to investigate the mechanism of Pingyangmycin sclerotherapy for cavernous hemangiomas. METHODS: Totally 9 rabbits (one as the control) were selected to be injected with Pingyangmycin and sodium morrhuate into the auricularis posterior vein, and then these veins were examined histologically 2, 7, 14 and 21 days after injection respectively. RESULTS: After injection, Pingyangmycin nonspecifically made the endothelia and veins defective, and induced proliferation of endothelial cells and smooth muscle cells after 7 days. Further, the veins became sclerostenosis 21 days after injection. On the other hand, sodium morrhuate caused thrombosis quickly and the vascular cavity disappeared. Finally, the thrombus became fibrogenesis. The local skin could be observed swollen and necrosis. CONCLUSION: Pingyangmycin is a better sclerosant for the therapy of cavernous hemangioma. PMID- 12539411 TI - [An experimental study of effects of injection of pingyangmycin on activities of enzyme of endothelial cells]. AB - OBJECTIVE: The objective of this study is to investigate the mechanism of acute trauma of endothelium after injection of Pingyangmycin. METHODS: A total of 42 adult Wistar rats were divided randomly into experimental groups (24), positive control groups (12) and normal control group (6), and the rats in the experimental or the positive control groups were injected with Pingyangmycin or sodium morrhuate into the mesenteric veins respectively, and then the enzyme activity of hepatic endothelial cells and hepatocytes was examined histochemically 0.5, 1, 2, 4, 8 and 24 hours after injection. RESULTS: After injection, the activity of 5'-nucleinase(5'-N) in hepatic endothelial cells and that of lactic dehydrogenase (LDH) and succinodehydrogenase (SDH) in hepatocytes decreased gradually, but that injected Pingyangmycin was less than that injected sodium morrhuate. CONCLUSION: The trauma caused by Pingyangmycin is less than that caused by sodium morrhuate, but the reaction of trauma caused by both is nonspecific. PMID- 12539412 TI - [IL-2 gene and cisplatin combined therapy for head and neck squamous cell carcinoma]. AB - OBJECTIVE: The objective of this study is to assess the efficacy of IL-2 gene and cisplatin combined therapy on murine HNSCC models. METHODS: HNSCC tumors were established in murine models. The sizes of tumors were measured before and after combined IL-2 and cisplatin treatment, and the results were evaluated by comparing different single treatment groups and the control IL-2 expression was measured after IL-2 gene transfer. Cytotoxic T-lymphocyte (CTL) and natural killer (NK) assays were also performed. RESULTS: HNSCC tumor growth was significantly inhibited following combined IL-2 gene and cisplatin therapy, when compared with that of the control. The levels of IL-2 protein expressions were increased in combined and single IL-2 treated groups. The activities of cytotoxic T-lymphocytes and natural killers in both the combined treatment group and the IL 2 treated group are higher than those of the controlled group. CONCLUSION: Combined IL-2 gene and cisplatin therapy can significantly inhibit HNSCC tumor growth in the murine models, and efficiently induces anti-tumor immune reaction of hosts. PMID- 12539413 TI - [Effects of immune bovine whey on sucrose-dependent adherence of Streptococcus mutans]. AB - OBJECTIVE: The aim of this experiment is to explore the possible anti-carious mechanisms of immune bovine whey. METHODS: The immune milk was collected from cows immunized with cell associated glucosyltransferase(GFT) over-expression strain B-29-33 of Streptococcus mutans(S. mutans) MT8148. The control milk was obtained from non-immunized cows. The immune and control bovine whey were prepared by removing fat and casein. The effects of two kinds of bovine whey on sucrose-dependent adherence of S. mutans MT8148 were examined by using TTY media with five different volumes of the immune and the control milk, 100, 300, 500, 700 and 900 microliters respectively. The adherent rates were determined by reading the optical density. The adherent morphology was observed by using a scanning electron microscope(SEM). RESULTS: There was no significant difference in adherence of S. mutans MT8148 cells to the smooth glass surface among five doses with those of the control whey. The immune bovine whey inhibited the adherence of S. mutans. The adherent percentage was 84.37% at 100 microliters and decreased by 58.59% at 900 microliters. The inhibition was does-dependent. The difference of adherent morphology between the immune and control whey was noticeable at 900 microliters. The SEM revealed that cells of S. mutans MT8148 with the control whey were covered by large quantities of gelatinous material, but cells with the immune whey were clearly distinguished. CONCLUSION: The immune whey has inhibitory potential against the sucrose-dependent adherence of S. mutans. PMID- 12539414 TI - [Expression of type I collagen mRNA in rat molar periodontal ligament under normal bite force]. AB - OBJECTIVE: The main component of the periodontal ligament(PDL) is collagen fiber, especially type I collagen, and collagen plays an important role in PDL. The aim of this study is to observe the expression of rat PDL mRNA under normal bite force and to investigate the molecular mechanisms of these changes in type I collagen mRNA. METHODS: 40 male Wistar rats were used, and animals were intra cardiac perfused with a solution of 4% polyformaldehyde under anesthesia. Dissected mandibles were immersed in the same fixation for 6 hours and subsequently decalcified in EDTA. The demineralized specimens were embedded in paraffin and cut into slices with thickness 5 microns. The probe was synthesized and labeled with digoxigenin. Expression of type I collagen mRNA was measured by using in situ hybridization(ISH). RESULTS: Under the normal bite force, the mRNA expression of type I collagen was very strong on the whole, including the alveolar bone side, the root side and the area between them. Positive signals were located mainly in the cytoplasm and some in the nuclei. But the mRNA expression of type I collagen still had spatial characteristics. The signals in some fibroblasts were apparently stronger than those in other fibroblasts in the apical 1/3 fragment of the roots. The signal of type I collagen mRNA was strong near the root sides. The expression signal on the proximal alveolar walls was strong, however, on the distal alveolar wall, there was no expression. CONCLUSION: The expression of type I collagen mRNA is closely related with bite force. PMID- 12539415 TI - [Study of the changes and effect of VEGF and bFGF in early stage of maxillofacial blast injury in rabbits]. AB - OBJECTIVE: The aim of this study is to determine the content of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in the wound fluid during maxillofacial soft tissue blast injury, and to evaluate their effects on angiogenesis of wound healing. METHODS: The rabbit model of maxillofacial blast injury was made by KTY-04 blasting cap. The content of VEGF and bFGF in wound fluid which was collected by using polying alcohol (PVA) sponge, were detected by ELISA. The distributions of VEGF and bFGF in the wound tissue were observed by using the techniques of in situ hybridization and immunohistochemistry. RESULTS: In the wound fluid of maxillofacial blast injury, the content of VEGF was rising steadily at the first week after injury. Comparing with natural sera, it showed difference from the first day after injury, and showed obvious different from the third day (P < 0.01), on which the content was 2.9 ng/ml +/- 2.7 ng/ml. It reached the peak on the seventh day after injury. The bFGF level in wound fluid was 565 pg/ml +/- 436 pg/ml, the peak value was gained at the sixth hour after injury. In the following days, it declined close to the serum level three to five days after injury, and then moved up slightly on the seventh day after injury. The expression of VEGF was located in epithelial cells, vascular endothelial cells and some chronic inflammatory cells. The mRNA transcription of bFGF was present in fibroblasts, endothelial cells and macrophages. CONCLUSION: VEGF and bFGF take part in two angiogenic cascades with synergistic action in maxillofacial blast injury. The bFGF stimulates the initial angiogenesis and induces the production of VEGF. The subsequent and more prolonged angiogenic stimuli can be mediated by VEGF. PMID- 12539416 TI - [Effects of clasp designs of Kennedy Class II RPD on the displacement of alveolar bone around the abutment tooth--an under load hologram study]. AB - OBJECTIVE: This article is to investigate the effects of different clasp designs of Kennedy class II RPD on the displacement of alveolar bone around the abutment tooth. METHODS: This experiment was carried out by using a separate human mandible. Five clasp designs were examined in this experiment, including RPA, RPI, modified RPI (mRPI), three-arm clasps and buccal wrought wired three-arm clasps. Hologramatic measurement was applied to record the different effects of these five designs of clasps on buccal and lingual alveolar bone. RESULTS: The RPI clasp was found to cause the smallest bone displacement. Generally, the clasps with the mesial occlusal rest (RPA, RPI, mRPI) caused smaller bone displacements than those with distal occlusal rests. The bar clasps brought about evenly spread bony displacements, however, the circumferential clasp caused highly concentrated displacement around apical site of the abutment tooth. CONCLUSION: The results suggest that mesial occlusal rests and bar clasps are preferable designs in this kind of prostheses. PMID- 12539417 TI - [Ultrasonic measurement of the elastic properties of goat condyles]. AB - OBJECTIVE: The purpose of this experiment is to investigate the elastic properties of goat mandibular condyles and to provide basic constants for biomechanical study of temporomandibular joint. METHODS: A fresh goat mandible was adopted and the standard condyle specimen of 4 mm x 6 mm x 10 mm was obtained from it. The structure density of the condyle was determined according to Archimede's principle. The transmitting velocities of ultrasonic wave in different directions of goat condyles were measured by using the ultrasonic system, which was composed of a transmitting transducer of 1.25 MHz, a receiving transducer of 2.5 MHz, an ultrasonic analyzer (5052UA) and a digital storage oscilloscope (HP54601A). The elastic coefficients and technical constants could be calculated by elastic equations. RESULTS: The structural density of the sample and the wave velocities in different directions of the condyle were both obtained by direct measurement. And then, nine elastic coefficients and all of the technical constants of orthotropic condyles were calculated. CONCLUSION: 1. It is practicable to measure elastic properties of the condyle by using ultrasonic technique. 2. Goat condyles of the mandible are a type of anisotropic and linearly elastic material. 3. The direction perpendicular to the transverse ridge of the goat condyle has the highest Young's modulus. As to shear modulus, the direction along the transverse ridge shows the highest value. PMID- 12539418 TI - [Effects of fluid shear stress on PGE2 synthesis of primary osteoblast-like cells in rats]. AB - OBJECTIVE: The purpose of this study is to study the function of PGE2 during the signal transduction in which fluid shear stress regulates the proliferation and differentiation of osteoblast-like cells. METHODS: The isolated rat primary osteoblast-like cells were exposed to fluid shear stress 0.12 mN/cm2 for 5, 10, 15, 30, 60 and 120 minutes respectively in a flow chamber. The release of PGE2 was examined. RESULTS: After exposure to fluid shear stress, the PGE2 synthesis of rat primary osteoblast-like cells increased significantly (P < 0.01) when compared with the control. After 10 minutes, the release of PGE2 began to increase significantly (P < 0.01) and the effect was maximal after 60 minutes (P < 0.01). CONCLUSION: PGE2 pathway may be one of the signal-transduction pathways which can transduce the fluid shear stress into osteoblast-like cells and then stimulate the bone remodeling. PMID- 12539419 TI - [A study on transmission of pathogenic bacteria of rampant caries from mothers to children]. AB - OBJECTIVE: In order to prevent rampant caries in children, the transmission from mothers to children of pathogenic bacteria of rampant caries will be investigated in this study. METHODS: By using fingerprint technique, the genetic diversities of Streptococcus mutans(S. mutans) were examined respectively in 10 children with rampant caries, 10 children without rampant caries S. mutans, 10 2-year-old caries-free children, and their mothers. The levels of S. mutans in saliva of mothers were also examined. For fingerprinting, chromosomal DNA of isolates obtained from plaques of these children and from saliva of their mothers were digested with restriction endonuclease HindIII and examined by electrophoresis on 0.7% agarose gel. DNA fingerprints were obtained and analyzed for genotypic similarities. RESULTS: The DNA fingerprint patterns showed that the proportion of children whose genotypes of S. mutans matched their mothers in the children with rampant caries was 70%, which is similar to that in the children without rampant caries and the caries-free children (60% respectively). The number of distinct genotypes of S. mutans harbored in children with rampant caries was, on average, greater than the number of genotypes present in children without rampant caries and caries-free children (1.7, 1.2 and 1.1 respectively, P < 0.05). Mothers of children with rampant caries had similar levels of S. mutans in saliva as those mothers of children without rampant caries and caries-free children. However, mothers of children with rampant caries had significantly higher DMFT scores and higher levels of Streptococcus sobrinus (S. sobrinus) in saliva than the mothers of caries-free children did. CONCLUSION: In all child groups, mothers were probably the main source of infection with S. mutans. Rampant caries in children is associated with DMFT scores and levels of S. sobrinus in saliva of mothers. In the initiation of rampant caries experienced by a child, the transmission of pathogenic bacteria from mothers to children may be the main source. PMID- 12539420 TI - [A study on bacterial contamination of dental handpieces]. AB - OBJECTIVE: The aim of this study is to evaluate the effects of different factors, including concentration of the bacterial suspension, number of stops set on handpieces, and different disinfectants, on bacterial contamination of handpieces and their clinical significance. METHODS: Bacterial contamination of 20 handpieces were analyzed in different concentration of bacterial suspension, number of stops set on handpieces and after using different out surface disinfectants. RESULTS: Statistical increment (P < 0.05) of handpieces inside contamination was observed relevant to increment of the bacterial suspension concentration. No statistical relevance was seen between handpiece inside contamination and the number of stops set on handpieces(P > 0.05). Disinfection with different disinfectants significantly decreased out surface contamination(P < 0.05), but no statistical change of bacteria (P > 0.05) was observed inside handpieces. CONCLUSION: As the concentration of bacterial suspension is the key factor affecting the inside contamination of handpieces, oral cavity cleanness is essential before dental therapy. The surface disinfection of handpieces is necessary after dental therapy, but cannot prevent cross-contamination between patients. PMID- 12539421 TI - [The clinical application and modification of the Quad Helix appliance]. AB - OBJECTIVE: The aims of this study are to investigate the working mechanism, the characteristics, the clinical application and the suggested modification of the Quad Helix appliance. METHODS: A 7-year-old, female patient with Pierre-Rokin syndrome, who was preformed with palatorrhaphy at 21-month-old treated by using a Quad Helix appliance for one year. RESULTS: After one-year treatment, the wide between the maxillary first molars increased 9.65 mm, and the wide between maxillary canines increased 5.20 mm. The wide between the mandibular first molars also increased 3.60 mm, however the wide between mandibular canines decreased 5.20 mm. CONCLUSION: The Quad-helix appears to be a successful Orthodontic appliance to expand the narrow maxillary or mandibular arches. PMID- 12539422 TI - [Interleukin-6 levels in the gingival crevicular fluid before and after periodontal treatment]. AB - OBJECTIVE: The aim of this study therefore is to determine if periodontal scaling and root planning could affect the IL-6 level in the GCF of teeth with periodontitis. METHODS: 12 teeth from 12 patients with severe periodontitis were clinically examined and sampled before therapy. 2 weeks after the periodontal scaling and root planning, the same 12 teeth were examined and sampled again. The IL-6 levels in GCF were detected using ELISA method. There are some evidences that the interleukin-6 (IL-6) levels in the periodontal tissue and gingival crevicular fluid (GCF) of periodontitis sites are significantly higher than that of healthy periodontal sites. RESULTS: The IL-6 levels in GCF from the teeth declined and the periodontal clinical indices of these teeth also apparently dropped down. CONCLUSION: Periodontal treatment, including periodontal scaling and root planning could affect the IL-6 levels in GCF of teeth with periodontitis. PMID- 12539424 TI - [Comparative study on self-made denture adhesive paste and fixodent in improving chewing efficiency of dentures]. AB - OBJECTIVE: The objective of this study is to test whether the self-made denture adhesive paste can improve the chewing efficiency of complete denture. METHODS: The abilities to improve chewing efficiency of two different materials, including self-made and Fixodent denture adhesive paste, will be investigated in this experiment. 24 patients were randomly selected, who wore complete dentures or mucous membrane supporting removable partial dentures because of the loss of most teeth. Identical test method was adopted by chewing peanuts. By employing a spectrophotometer, the absorbency values of chewed peanuts suspension before and after the use of self-made denture adhesive pasties were measured, and the differences could be obtained by calculation. The differences indicated the relationship between denture adhesive paste and chewing efficiency. Then the results of two materials would be compared and analyzed. RESULTS: The statistical analysis revealed that both self-made denture adhesive paste and Fixodent resulted in remarkable difference in the absorbency values before and after use (P < 0.01), however, there is no obvious difference in improving the chewing efficiency between the self-made and Fixodent denture adhesives (P > 0.05). CONCLUSION: The self-made denture adhesive paste is the same as Fixodent in improving the chewing efficiency of denture. PMID- 12539423 TI - [Effects of psychosocial factors on recurrent aphthous ulcer]. AB - OBJECTIVE: The aim of this study is to investigate the effects of patients' psychosocial background on the recurrence of aphthous ulcer (RAU). METHODS: A total of 105 patients with RAU and 100 normal subjects of the control group were evaluated for psychosocial background. All subjects were required to complete the EPQ questionnaire, the SCL 90 questionnaire, and the enquired data were analyzed by using chi-square test. RESULTS: Special personality and social factors in RAU patients might be reasons leading to the occurrence of this disease (P < 0.01). CONCLUSION: Psychosocial disorder caused by stresses of social life events on the people with special personality could impact the occurrence of RAU. PMID- 12539425 TI - [Orthodontic diagnosis and treatment of unilateral posterior cross-bite]. AB - OBJECTIVE: The aim of this study is to investigate the clinical diagnosis and treatment of unilateral posterior cross-bite and to provide information for correct treatment of this disease. METHODS: The X-ray examination of patients' heads in the posterior-anterior position, analysis of pantomorphic films and model measurement were used for the diagnosis of unilateral cross-bite. According to these data, three types of unilateral posterior cross-bite would be classified, and different orthodontic appliances would be applied in the following treatment, including separate plate removable expansion appliances, Quad-helix appliances, narrowing dental arch and inter-arch elastics. RESULTS: A total of 78 cases of unilateral posterior cross-bite were treated by different techniques in a manner consistent with the classification. All of the clinical results of treatment were satisfactory and successful. CONCLUSION: If diagnosis is correct, and suitable treatment methods are chosen, unilateral cross-bite can be treated successful. PMID- 12539426 TI - [X-ray changes of temporomandibular joints after indirect trauma]. AB - OBJECTIVE: The aim of this study is to investigate the structural changes of goat's temporomandibular joints (TMJs) after indirect trauma. METHODS: The trauma to the goat's TMJ achieved under an impact to the right and left mandibular angles using a self-made device. These goats were killed after 2 hours, 7 days, 1 month and 3 months. The goat's TMJs were examined using tomography before being killed, and also checked by using a histological method. The normal temporomandibular joints were treated as the control. RESULTS: 1. The X-ray tomograms: In the early post-trauma stage, the articular cavities were dimmed, and the joint spaces were decreased; the articular surfaces were not smooth, and the structures of TMJ were not clean. Moreover, the stations of the condyle were moved from middle of the fossa to the posterior fossa in the later stage. 2. The histological examination: The condyles exhibited eroded surfaces with osteophytes, the discs were thinned or perforated, and the temporal surfaces were thick-end and the linings were separated. CONCLUSION: The indirect trauma on TMJs of goats can cause the TMJ internal derangement and the osteoarthrosis of TMJs. The X-ray tomography is a very useful method in the long-term monitoring of bone changes in TMJs after trauma. PMID- 12539427 TI - [MRI manifestation of temporomandibular joints after indirect trauma]. AB - OBJECTIVE: The objective of this study is to investigate the structure changes in temporomandibular joints (TMJs) of goats after indirect trauma. METHODS: The trauma of TMJs of goats achieved under an impact to the right and left mandibular angles with a self-made device. These goats were killed after 2 hours, 7 days, 1 month and 3 months. The TMJs of goats were examined using MRI before killed and checked by histological method, the normal temporomandibular joints were treated as the control. RESULTS: The results of MRI: in the early post-trauma stage, the condylar articular surface was partially lost, and the discs were displaced and ruptured. Further, the hemathrosis of the TMJ and the decreased joint spaces were also observed in the eroded, eburnous surface of the condyle and the deformed disc. In the later stage, the surfaces of condyles were eroded and eburnated, and the discs were deformed. CONCLUSION: The indirect trauma on the TMJs of goats can cause TMJ internal derangement and TMJ osteroarthrosis. MRI cab be used to investigate both hard and soft tissue injuries of the temporomandibular joint, and is appreciated in understanding the pathogenesis of indirect trauma on the TMJ. PMID- 12539428 TI - [Impact of interstitial irradiation with 32P glass microspheres on cellular apoptosis of mice with the solid tumor S180]. AB - OBJECTIVE: The aim of this study is to observe apoptotic changes of tumor cells in mice with the solid tumor S180 after interstitial irradiation with 32P glass microspheres. METHODS: Twenty mice with solid tumor S180 were divided into four groups. The control group was given normal saline. The experimental groups were given different doses of 32P glass microspheres (50, 150, 450uci per mouse) using interstitial implant. The terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method was used to determine the apoptotic cells. RESULTS: The mean values of the apoptotic indexes of these groups were respectively 0.39, 0.41, 0.59 and 0.95. The apoptotic indexes of the second and the third experimental groups were significantly higher than that of the control group (P < 0.05). CONCLUSION: The interstitial irradiation with 32P glass microspheres can enhance the activity of apoptotic cells of solid tumor S180, and the effect is dose-dependent. PMID- 12539429 TI - [Apoptosis induced by interstitial irradiation with 32P glass microspheres combination with hyperthermia in mouse solid tumor S180]. AB - OBJECTIVE: The aim of this study was to observe the changes of tumor cell apoptosis after interstitial irradiation combination with the hyperthermia treatment. METHODS: Twenty mice with solid tumor S180 were divided into four groups, including a control group which was given normal saline, and three experimental groups which were respectively applied with a hyperthermia treatment (bath water), interstitial irradiation with 32P glass microspheres, and the combined treatment of hyperthermia and interstitial irradiation. The terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method was used to determine the final apoptotic cells. RESULTS: The mean values of apoptotic indexes of these four groups were respectively 0.39, 0.53, 0.59 and 0.91. The apoptotic indexes of these experimental groups were significantly higher than that of the control group (P < 0.05). The apoptotic index of the combined treatment group was significantly higher than those of the hyperthermia group and the interstitial irradiation group (P < 0.05). CONCLUSION: Both the interstitial irradiated with 32P glass microspheres and hyperthermia (bath water) can induce the apoptosis of mouse solid tumor S180. It seems that there is a synergistic induction of apoptosis between interstitial irradiation and hyperthermia. PMID- 12539430 TI - [Shear bond strengths of four types of adhesive resin agents to GI II glass infiltrated ceramic]. AB - OBJECTIVE: The shear bond strength of four adhesive resins bonded to GI II glass infiltrated alumina ceramic was investigated in this experiment. METHODS: A total of 40 ceramic blocks and 40 tooth blocks were prepared and divided into 4 experimental groups, including group A (CereDual and silicoating), group B (Calibra and silicoating), group C (Panavia) and group D (Super C&B and silicoating). The shear strength of four different bonding systems to dentin was tested respectively. The data were statistically analyzed. RESULTS: More than 13 MPa of bonding strength was achieved using the traditional Bis GMA bonding resin (groups A and B), and more than 16 MPa was obtained using the improved Bis GMA bonding resin (group C and D). There was statistical difference between groups A, B and group C, D. CONCLUSION: It can be implied that an ideal bonding can be obtained by using different bonding materials and processing the dentin surface with the suitable conditioner. PMID- 12539431 TI - [A survey of oral health status of the elderly individuals in Chengdu of China]. AB - OBJECTIVE: The aim of this survey is to investigate the oral health status of the oldsters over 65-year-old in Chengdu of China. METHODS: The oral health status, including oral hygiene, caries, periodontal diseases and the tooth loss of 468 elderly individuals over 65 years old were surveyed and analyzed. RESULTS: It was found that all these older individuals brushed their teeth everyday, but most of them had tooth decay, gingivitis and periodontitis. About 7.47% of them were edentulous, and 2.56% of them were half edentulous. The masticatory ability of most of them declined and the chewing ability of them was low. CONCLUSION: The oral health status of these elderly individuals in Chengdu is poor. It is therefore necessary to improve the health status of this population. PMID- 12539459 TI - [Effects of bite force lost on the expression of IL-6 in the rat periodontium]. AB - OBJECTIVE: In order to explain the molecular mechanism of periodontium remodeling the expression of IL-6 in the rat periodontal ligament fibroblasts (PDLC) and the alveolar bone osteoblasts under normal and bite force lost was studied. METHODS: To establish a rat model of different bite force for the study, left maxillary molars of rats were extracted and the left mandibular molar area was used as the bite force lost. The animals were sacrificed after 6 hours, 1, 2, 3 days and 1, 2, 3, 4 weeks. The prepared tissue specimens were processed for the study of the change in the histologic morphology and the expression of IL-6 protein in PDLC and osteoblasts with HE staining and immunohistochemistry techniques. RESULTS: In the group of bite force lost, the loosely arrangement of periodontal ligaments and the resorption of alveolar bone were observed in histological study, the expression of IL-6 in PDLC and osteoblasts was enhanced apparently than that in the group of normal bite force by using immunohistochemistry techniques. In the group of lost bite force, the expression of IL-6 in PDLC enhanced with time regularly: The expression of IL-6 was strengthened after 3 days, and reached the peak after 2 weeks, then, decreased after that and restored to normality in 4 weeks. While in osteoblasts of the group of bite force lost, the expression of IL 6 was strengthened just after 2 weeks. CONCLUSION: Bite force lost induce the expression of IL-6 enhanced apparently in PDLC and osteoblasts, it suggests that IL-6 may play important roles in the effect of bite force on periodontium remodeling. It approaches initially the molecular mechanism of periodontium remodeling, demonstrates that the histologic morphology is closely related to the mechanical condition. PMID- 12539460 TI - [The devitalization of tumor suppressor genes in oral squamous cell carcinoma]. AB - OBJECTIVE: To study the expression of anti-oncogenes nm23, P16, P53 of squamous cell carcinoma within one sample. METHODS: A total of 27 cases of oral squamous cell carcinoma came from our the First Clinical College, China Medical University from 1997 to 1998, and none of them had the history of radioactivity therapy or chemical therapy. In them, 8 cases were in the early period, and 19 cases were in the late period. 15 cases were male and 12 cases were female. The samples were fixed in formaldehyde solution, embedded with paraffin, sliced in 5 microns, and analysed by ABC immunohistochemistry staining. The first antibodies were anti P16, anti-nm23, anti-P53 respectively. The criteria of judgment were the following: For nm23, the case that cytoplasm or nucleus were buffy particles or ball was positive. For P16, the case that nucleus were brown was positive, while the case with the light brown cytoplasm and unstained nucleus was negative. The stained nucleus of interstitial cells were treated as the control. For p53, the positive protein product was assembled in the carcinoma cell nucleus, so the nucleus with buffy or brown particles was positive. According to the criteria, 5 high power sights were selected from each case, more than one hundred cells were counted. The case that the positive cells was more than 30 percentage was high expression, and less than 30 percentage was low expression. Two doctors completed the procedure respectively, and only the same results could be qualified. RESULTS: In the 27 cases, the devitalization of nm23 and p16 was showed in low expression, p53 in high expression. The devitalization rate: p16 was 66.7%, nm23 was 48.1%, and p53 was 25.9%. In 14 cases, combined mutation of two inhibitory genes were seen. 9 cases were p16 and nm23, among them, 4 cases had transferred to cervical lymph nodes (In all the cases, there were 6 cases who had cervical lymph node transference). CONCLUSION: p16 genetic changes are more often detected in oral squamous cell carcinoma than the other genetic changes; In a large portion of cases, there were combined genetic changes instead of single gene mutation. PMID- 12539461 TI - [The expression of PCNA and NOR in carcinogenesis procession of hamster buccal pouch mucosa]. AB - OBJECTIVE: To investigate the expression of proliferating cell nuclear antigen (PCNA) and nucleolar organizer regions (NOR) with the carcinogenesis model of hamster buccal pouch mucosa. METHODS: 48 Syrain hamsters with 6-8 weeks old and 70-80 g weight were selected. The material used for inducing cancer is 0.5% DMBA (7,12-dimethylbenzanthracene) in acetone. 0.5% DMBA was applied in the right buccal pouch of the hamster 3 times a week for 12 weeks. The control received no treatment. The time for collecting specimens was 3, 6, 9 and 12 weeks. The specimens were immediately fixed with 10% formalin, dyed with HE, and then two doctors major in histopathology evaluated with the WHO criterion (1986). The wax of immunohistochemical specimen was cut and placed on the APES pieces of glass, treated with 50 degrees C for 2 hours and preserved under the indoor temperature. The PCNA was examined with the LSAB technique of immunohistochemistry. The company called Zymed provided the MonAb. The positive control was a sample of human inflammatory hyperplastic lymphoma, and the negative was PBS replacing the MonAb. NOR count was determined in a method of silver nitrate staining. The test condition should be at 45 degrees C without light. RESULTS: 1. AgNOR was usually located in the basal-cell layer of normal epithelium, mainly in a single form. High dysplasia was mainly in an aggregated form, however, carcinoma in infiltration appeared to be in a mixed form. 2. PCNA expression was similar to that of the normal control in hyperplastic epithelia (3 weeks) at a moderate level in dysplasia epithelia (6-9 weeks). PCNA expressed highly in squamous cell carcinoma. The expression of the PCNA in line with malignant progress became stronger increasingly. The positive cell was perceivable in all epithelia of high dysplasia. CONCLUSION: PCNA has been significantly positive and correlative with AgNOR (r = 0.635, P < 0.001), however, the film of PCNA is much clearer and the practical value is greater. PMID- 12539462 TI - [Comparison of mechanical properties of laser-welded wrought and cast titanium]. AB - OBJECTIVE: This study was aimed to compare the mechanical properties of laser welded wrought and cast titanium. METHODS: According to relevant reports and clinical demands, the specimens of TA2 were machine milled or cast into certain sizes using LZ centrifuge-vacuum-pressure titanium casting machine. This experiment involved four groups including the laser-welded wrought titanium group, the laser-welded cast titanium group, the wrought titanium group and the cast titanium group. There were 6 complete specimens in each group and the specimens were ultrasonically cleaned with ethyl and distilled water before they were welded with home-made Nd:YAG apparatus, and comparisons of mechanical properties were made among these groups. All tensile specimens were mounted on the Instron universal testing machine to test tensile loads, yield loads and absolute elongations were measured. The cross section of fractured surface was observed under S-2700 Hitachi SEM and Vickers' microhardness values in different areas were detected with AKASHI microhardness tester. RESULTS: It was showed that the tensile load of laser-welded wrought titanium made no difference from that of the original wrought titanium, but the yield load of laser-welded wrought titanium was bigger than that of the original wrought titanium. The tensile load and yield load of cast titanium were slightly bigger than those of wrought titanium. The tensile load and yield load of laser-welded wrought titanium made no statistic difference from those of laser-welded cast titanium. The absolute elongations of cast or welded titanium were lower than those of wrought or non welded titanium. The fractured surface of cast titanium under SEM appeared lath like structure and that of welded cast titanium honeycomb-like structure. The honeycomb-like structure of wrought titanium was more obvious than that of laser welded wrought titanium. The microhardness value of laser-welded cast titanium was slightly bigger than that of laser-welded wrought titanium. CONCLUSION: The mechanical property of laser-welded wrought titanium might be not apparently different from that of laser-welded cast titanium. PMID- 12539463 TI - [Effects of pulsed Nd:YAG laser irradiation on shear bond strength of composite resin bonded to porcelain]. AB - OBJECTIVE: To evaluate the shear bond strength of composite resin bonded to pretreated porcelain surfaces by use of a pulsed Nd:YAG laser irradiation and by use of 8% hydrofluoric etching. METHODS: Surfaces of porcelain were roughened with coarse diamonds until the opaque body porcelains were exposed. The samples were divided into four groups. Samples of the first group were etched with 8% hydrofluoric acid for 3 min by following the manufacter's instructions. Samples of the 2nd, 3rd and 4th groups were separately irradiated by a pulsed Nd:YAG laser of three energy parameters of 15 Hz, 40 mJ; 0.6 W; 15 Hz, 60 mJ, 0.9 W and 15 Hz, 80 mJ, 1.2 W different respectively, each specimen was irradiated for 1 min. Then, silane coupling agent was equally placed on the treated surfaces for 3 min, the activator was used on each surface. The adhesive was applied and light cured. The composite resin material was finally bonded to the treated porcelain surface, forming a 3 mm x 3 mm x 2 mm resin mass. Samples were then tested for shear bond strength between the treated porcelain and composite resin by using a SWD-10 electric universal testing machine with a crosshead speed of 1 mm/min until fracture. At the end of testing, the data was evaluated by using a two-way analysis of variance (ANOVA) comparing the shear bond strength. On the other hand, characteristic morphology of laser irradiated surfaces of porcelain was prepared and observed by a scanning electron microscopy. RESULTS: The shear bond strength of laser groups of 0.6 W, 0.9 W, 1.2 W and etching group was respectively 8.61, 14.07, 11.22 and 13.47 MPa. There was no significantly difference among groups of laser 0.9 W, 1.2 W and the etching group. This suggested that irradiated on surfaces of porcelain by Nd:YAG laser at the above laser energy could attain the same effectiveness as hydrofluoric acid etching. But the shear bond strength of laser 0.6 W group was significantly lower than the etching group. It suggested that shear bond strength would obviously decrease when laser energy was 0.6 W or lower than this parameter, and had no use in treatment on porcelain surfaces comparing to acid etching. SEM showed that porcelain surfaces irradiated by Nd:YAG laser exhibited rough, umbilicate, lava crater-like structure. This structure helped to produce mechanical retention between resin composite and porcelain. CONCLUSION: The pulsed Nd:YAG laser could replace etching method to pretreat the fracture porcelain surfaces for bonding with composite resin at appropriate energy parameters. PMID- 12539464 TI - [The effect of fibrin sealant on dental pulp for pulp capping in experimental dogs]. AB - OBJECTIVE: To investigate the pulp tissue reaction and reparative dentine formation of dog pulp capped with Fibrin Sealant (FS). METHODS: The fibrin was from a tisseel Kit (Immuno AG, Vienna, Austria) consisting of solution A (freeze dried human fibrinogen, factor XIII, Aprotinine) and solution B (thrombin powder, calcium chloride) with 0.02 ml from each for each tooth. The Ca(OH)2 paste (Calar, China) was served as the control. Forty nine cuspid, premolar and molar teeth in 4 adult dogs weighed 11-13 kg were used. Surgical anesthesia was obtained by muscle injection of 3% sodium pentobarbital (1 ml/kg). The pulp were exposed by using sterile high-speed rotary cutting instruments with water cooling. The pulps were treated with 0.02 ml fibrin or Ca(OH)2 paste respectively. The cavity was filled with zine phosphate cement. The animals were killed at intervals of 1, 4, 6, 9 weeks after surgery and perfused with phosphate buffered saline followed by 10% formalin in phosphate-buffered saline. The jaws were resected and the samples prepared by removing each experimental tooth in block, further fixing them by immersion in 10% formalin in phosphate-buffered saline. The specimen were processed for histologic examination. Serial sections (5 nm thick) were made, stained with hematoxylin and eosin, examined using a light microscope. RESULTS: 7 days after operation, the samples treated with the FS showed no hemorrhage and degeneration of pulp tissue and a small amount inflammatory cells near the wounds. The samples treated with Ca(OH)2 showed hemorrhage and degeneration (2/6) and a large amount inflammatory cells near the wounds. 28 days after operation, pulp health of teeth treated with FS and Ca(OH)2 was nearly recovered. Dentin bridges were observed in 2 out of 6 cases treated with Ca(OH)2. 42 days after operation, pulp health of teeth treated with FS and Ca(OH)2 was totally recovered. Dentin bridges were observed in 2 out of 7 cases of pulp treated with FS and four out of 6 cases treated with Ca(OH)2. 63 days after operation, pulp health of teeth treated with FS and Ca(OH)2 was soundly resumed. The dentine bridge was observed in 4 out 6 cases of pulp treated with the FS and in all 6 cases treated with Ca(OH)2. CONCLUSION: FS has no dentin inducing activity, but can enhance exposed pulp healing, so FS can be served as a promised pulp capping agent. PMID- 12539465 TI - [Determining the distribution of Mutans Streptococci in human dental plaque by monoclonal antibody against SA I/II and SPAa of Mutans Streptococci]. AB - OBJECTIVE: To explore the possibility of detecting the distribution of Mutans Streptococci in human dental plaque by Monoclonal antibody (WC2E10c, WC3A6d) against SA I/II and SPAa of Mutans Streptococci, which was prepared in our lab. METHODS: 60 subjects were divided into the experimental group and the control group. The distributions of Mutans Streptococci (serotype c and d) in dental plaques were detected in 60 subjects by the clonal blot technique. Positive colonies were examined by bacterial morphology, serology, biochemistry and SDS PAGE. RESULTS: Bacterial colony was significantly different between the experimental group and the control group with eye observation. Colonies of the experimental group were deep brown and easily differentiated, but colonies of control group were white. Brown colonies were proved to be Mutans Streptococci (serotype c and d) by using assay of bacterial morphology, serology, biochemistry and SDS-PAGE, but white colonies were not. CONCLUSION: McAbs (WC2E10c, WC3A6d) were shown high specificity in Mutans Streptococci (Serotype c and d) of human dental plaques. PMID- 12539466 TI - [Establishment of a GC and GC/MS method for analysis of fatty acids in human dental plaque]. AB - OBJECTIVE: To study the composition of fatty acids in human dental plaque, a Gas Chromatography (GC) and Gas Chromatography/Mass Spectrometry (GC/MS) method was developed. METHODS: The C17:0 was internal standard and the C16:0 was test standard. Samples of plaque were methanolysis to be fatty acids methyl ester before detection. GC with OV-1 separated the sample for a quantitative analysis. The column size was 25 m x 25 mm. The temperature program was from 80 degrees C to 260 degrees C with 12 degrees C/min. The sample separation was carried out at fused silica capillary column with OV-1 for qualitative analysis by GC/MS. The column size was 23 m x 0.23 mm. The temperature program was from 40 degrees C to 240 degrees C at 6 degrees C/min. RESULTS: The linear regression equation was y = 0.0138X-0.1182, r = 0.9945, recovery was 90.18%-103.6%, CV intro-day was 2.60, 1.61, 2.20; CV inter-day was; 2.71, 1.69, 1.61. The lowest limit of detection was 0.6 ug. The GC and GC/MS diagram showed that internal standard and fatty acids of plaque have good separation. CONCLUSION: The method is simple, rapid accurate, and is useful to carry out qualitative and quantitative analyses of fatty acids in dental plaque. PMID- 12539467 TI - [Impacts of clinicopathologic parameters on lymphatic metastasis of oral squamous cell carcinoma]. AB - OBJECTIVE: To evaluate the effects of clinicopathologic parameters on lymphatic metastasis of oral squamous cell carcinoma. METHODS: One hundred and ninety-three patients who underwent therapy in our department were included in this study. Ten parameters (T and N stages, location, patterns of invasion, thickness, lymphocyte infiltration, karyokinesis count, degrees of differentiation, degrees of cell keratinization and nuclear polymorph) were evaluated to determine their effects on lymphatic metastasis of oral carcinoma using univariate and multivariate analysis. RESULTS: By using the univariate analysis, five parameters (T and N stage, pattern and depth of invasion, lymphocyte infiltration) showed significant impacts on pathologic nodal metastasis. From multivariate analysis, however, only patterns of invasion were independent predictors of metastasis. Also, patterns of invasion were the only significant factors that influence the post-operative metastasis besides pattern of neck treatment. CONCLUSION: The pattern of invasion is the most important parameter that influences the metastasis of oral squamous cell carcinoma. PMID- 12539468 TI - [Computer-aided compose panoramic arthroscopic images of the temporomandibular joint]. AB - OBJECTIVE: The computer-aided image processing method is established to compose arthroscopic images of temporomandibular joints(TMJs). METHODS: Arthroscopic images were input directly into a personal computer and recorded at a compact disk. By using the software- Photoshop 5.0 for Window 95, the images were edited and adjusted to form sagittal and/or coronal panoramic images of articular surfaces. RESULTS: The normal sagittal and coronal two-dimensional integrated panoramic arthroscopic images of TMJ were obtained, and qualities of these images were satisfied. CONCLUSION: The arthroscopic panoramic images established by using this method can demonstrate integrated structures of articular surfaces directly. PMID- 12539469 TI - [Quantitative and immunohistochemical analysis of endothelin-1 in oral submucous fibrosis]. AB - OBJECTIVE: This experiment is carried out to investigate roles of endothelin-1 (ET-1), a potent vasoconstrictor and mitogenic peptide, in fibrosis and collagen production. METHODS: 30 cases of oral submucous fibrosis(OSF), 10 cases of oral lichen planus (OLP) and 10 cases of normal control, were studied by means of SABC immunohistochemical method with polyclonal antisera to ET-1 and using image pattern analysis technique. RESULTS: 1. Oral mucosa from OLP and normal control expressed very little ET-1. By contrast, there was striking expression of ET-1 in oral mucosa from OSF; 2. The cellular content of ET-1 in OSF was significantly higher than that in normal control (P < 0.01), while the cellular content of ET-1 in the early and moderate stages OSF were significantly higher than that in the advanced stage(P < 0.01); 3. The cellular content of ET-1 in the early and moderate stages OSF were significantly higher than that in OLP(P < 0.01); 4. There was a significantly positive correlation between cellular contents of epithelia and interstitial(P < 0.05). CONCLUSION: These findings suggest that cell-specific expression of ET-1 may play a role in pathogenesis of OSF. PMID- 12539470 TI - [Comparative research on forms of dental and palatal arches between adults and children after rapid maxillary expansion]. AB - OBJECTIVE: To study changes of dental and palatal arch forms between adults and children after rapid maxillary expansion (RME). METHODS: Trans-arch and trans palatal widths, and molar and palatal angles on plaster models of 47 children and 47 adults were measured pre- and post-treatment with RME. RESULTS: 1. Trans-arch width: Increases of trans-molar and trans-cuspid widths were significantly greater in the child group than those in the adult group, except first and second premolars. In the child group, there was no significant difference of increased widths between posterior teeth and cuspids. But, in the adult group, increased widths in trans-arch of first molars, first and second premolars were significantly greater than those in cuspids. 2. Transpalatal width: Trans-palatal increases at mid-palates and the apex of palates were significantly greater in the child group than those in the adult group. 3. Molar angle: In the child group, no difference existed in the molar group between pre- and post-treament. But, in the adult group, statistical difference existed between pre- and post treatment. Prior to RME, the molar angle was the same between the child group and the adult group. The molar angle was significantly greater in the adult than that in the child group after RME. 4. Palatal angle: The changes were not of statistical significance in child palatal angles between pre- and post-RME. Significant differences existed in the adult group between pre- and post-RME. Increases of palatal angles were significantly greater in the adult than those in the child group. CONCLUSION: Significant differences existed between the child and the adult group in forms of dental arches and palatal arches, which focused on the changes of molars and palatal angles after RME. PMID- 12539471 TI - [Clinical retrospect on autoimplantation of traumatically dislocated teeth]. AB - OBJECTIVE: A total of 48 cases with autoimplantation of traumatically dislocated teeth were retro spected as clinical references. METHODS: Several indexes were recorded in details, for instance, tooth dislocation time, operation and postoperative management and patients were followed up periodically. RESULTS: Tooth dislocating intervals from trauma to implantation were from 0.5 to 48 hours. Therapeutic effectiveness was graded as perfect in 37 teeth (61.7%), good in 18 (30.0%) and poor in 5 teeth (8.3%), in which 3 teeth were extracted within 1 year. Three-year success rate was 83.0%, and five-year success rate was 61.7%. It was judged 20 teeth being as periodontal membranous healing, 35 bony healing and 5 fibrous healing. Pulpal vitality tests were positive in 23 and negative in 37 teeth. Root canal therapies were conducted in 20 teeth. CONCLUSION: Carefully protecting periodontal membranes, keeping good fixation, preventing from infection and avoiding occlusal injuries are important factors for successful implantation of the dislocated teeth. Periodical examination would be benefit to the long term prognosis. The correct methods for dislocated tooth preservation should be treated as the important part of health education. PMID- 12539472 TI - [Diagnostic values of MRI in combination with DSA in vascular malformations of the head and neck]. AB - OBJECTIVE: To evaluate prospectively diagnostic values of magnetic resonance imaging (MRI) as the initial examination for vascular malformations of the head and neck, and establish a flowchart based on radiodiagnostic features. METHODS: 45 patients with vascular malformations of the head and neck were evaluated prospectively with MRI. MRI was followed by angiography in 15 patients, contrast enhanced CT in 15 patients. MRI was performed by using spin-echo pulse sequences with Philips Gyroscan NT 1.0 Teslaunit. RESULTS: In all cases, MRI demonstrated usefully, and there was no other single imaging modality that could provide information about internal structures and flow characteristics as much as MRI. It made differentiation between high-flow and low-flow lesions obvious, however MRI was inferior to angiography in delineation of feeder vessels. CONCLUSION: MRI can be used as the initial methods for diagnosis of vascular malformations in the head and neck, and could be combined with angiography in cases with high-flow or combined-flow lesions. PMID- 12539473 TI - [Construction of two eukaryotic expression plasmids pcDNA3/pacA and pcDNA3/pacP expressing surface protein antigen of Streptococcus mutans]. AB - OBJECTIVE: Two gene vaccines which encoded amino terminal and intermediate region in the PAc protein were constructed and the feasibility and safety was investigated, in order to obtain a highly effective and secure anticarious way by immunization. METHODS: The target gene fragments pac-A and pac-P encoded amino terminal and intermediate region of surface protein antigen from Streptococcus mutans were obtained by PCR amplification respectively. Amplification products were isolated by using low melting-temperature agarose and recoveried from the gel. Then the two fragments were digested by Kpn I and Xho I, and ligated to the Kpn I-Xho I-digested eukaryotic shuttle expression vector pcDNA3 to create reconstructed plasmids pcDNA3/pacA and pcDNA3/pacP respectively. The ligated DNAs were transformed into Escherichia coli XL1-Blue. Transformed colonies carrying eukaryotic expression plasmid either pcDNA3/pacA or pcDNA3/pacP were screened by colony hybridization in situ. Then the reconstructed plasmids were isolated and identified by restricted endonuclease digestions, Southern hybridization and DNA sequencing. RESULTS: 1. Two DNA fragments pac-A and pac-P of the predicted size 1.3 kb and 1.2 kb were obtained by PCR amplified. 2. Eukaryotic expression plasmids pcDNA3/pacA and pcDNA3/pacP had been reconstructed correctly. 3. The sequencing data also indicated that the pcDNA3 construct contained the correct sequence and orientation of the two target genes. CONCLUSION: These two plasmids may act as gene vaccines for further study. PMID- 12539474 TI - [Expression of eukaryotic expression plasmid pcDNA3/pacA and pcDNA3/pacP in mammalian cells]. AB - OBJECTIVE: Prior to the use of pcDNA3/pacA and pcDNA3/pacP in vivo, the transcription and expression products of these two eukaryotic expression plasmids in mammalian cells were detected. METHODS: The eukaryotic expression plasmids pcDNA3/pacA and pcDNA3/pacP were transfected into COS-7 cells respectively with liposome according to the manufacturer's protocol. In order to generate stable transfectants, after a 48 h incubation period in regular Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% heat inactivated fetal bovine serum, cells were treated with neomycin (1 mg.ml-1 of G418) and maintained under continuous selective pressure. G418-resistant colonies, which became visible after 2 weeks, were isolated and screened for expression of the vector-encoded protein. The control transfections were performed with the pcDNA3 vector and without recombinant as well as vector plasmid DNA. The mRNA transcriptions of the two insertional genes were detected by RT-PCR assay and their expression products were analysed by labelled-avidin-biotin enzymed-linked immunosorbent assay, flow cytometry and Western blotting. The cells transfected by pcDNA3 were used as the negative control. RESULTS: 1. The two eukaryotic expression plasmids could be correctly transcripted and translated under the control of the CMV immediate early promoter in mammalian cells. 2. The protein products could be detected in cell plasma, cell membrane and the culture supernatant. CONCLUSION: The two eukaryotic expression plasmids pcDNA3/pacA and pcDNA3/pacP can express protein products which were encoded by insertional gene pac-A and pac-P in mammalian cells. PMID- 12539475 TI - [Evaluation of cariogenic potential of Streptococcus mutans isolated from caries free and -active persons: adherence properties to saliva-coated hydroxyapatite]. AB - OBJECTIVE: In this study, authors used saliva-coated hydroxyapatite to simulate tooth surface in oral cavity to evaluate adherence of different Streptococcus mutans isolated from caries-free and -active persons to saliva-coated hydroxyapatite. METHODS: Streptococcus mutans isolated from people with different carious experiences were divided into two groups: the caried-free group (DMFS = 0) including 19 persons and the caries-active group (DMFS > or = 6) including 40 persons. Totally 105 genotype Streptococcus mutans strains were obtains, including 22 strains from the caries-free group, and 83 strains from the caries active group. The adhering properties of Streptococcus mutans (serotype c) strains were investigated, and different adhering properties were compared between the caries-free group and the caries-active group in order to find the correlationship between carious experiences and adhering properties of Streptococcus mutans. Saliva-coated hydroxyapatite was used to simulated tooth surface in oral cavity. Streptococci mutans were cultured in TPY liquid medium containing 3H-TDR in the same radioactive concentration. After being cultured in an anaerobic incubator for 18 hours, Streptococci mutans were radiolabelled by 3H TDR. Saliva-coated hydroxyapatite and bacterial liquid(BSA = 0.52) were mixed for 90 minutes. Samples were assayed by using liquid scintillation counter, and binding abilities of strains were evaluated by the count per minute(cpm). RESULTS: Binding abilities of strains were significantly different in two groups, Caries-active people harbored more Streptococci mutans with high binding abilities(cpm > 2000) than caries-free people did(P < 0.05). The binding abilities to saliva-coated hydroxyapatite were different between strains of differing genotypes isolated from the same person. Each person always harbored one or two strains with high binding abilities(cpm > 2000) in the caries-active group. CONCLUSION: 1. Different genotypes have different adhering properties. 2. In caries-active group, different genotype strains have synergic effects, and adhesion of high binding ability strains cause caries susceptibility. 3. The differences in binding properties of isolates might relate to differences in carious experiences. PMID- 12539477 TI - [Allele-specific chromosome 9p deletion in oral cancer]. AB - OBJECTIVE: The aim of this study was to investigate the relationship between loss of heterozygosity (LOH) on chromosome 9p and the pathogenesis of oral squamous cell carcinoma (OSCC). METHODS: A total of 24 human OSCC specimens were analyzed for LOH on chromosome 9p using 8 microsatellite markers by means of polymerase chain reaction. RESULTS: In 24 cases of OSCC, LOH on chromosome 9p was identified in 10 of 24 cases (41.67%) with at least one marker. The main LOH were found on 9p21 at locus D9S171 (21.05%) and D9S304 (10.00%). There were also a deletion on 9p22-23 at locus D9S168 (22.22%) and D9S162 (15.38%). However, there was no statistically significant correlation between LOH at these loci with such clinical parameters as pathological types, tumor size and lymph-node metastasis. Microsatellite instability (MSI) is rare for any of the 8 markers in 24 OSCC. No MSI could observed using the common criteria for defining MSI-its detection in two or more markers. CONCLUSION: We found that high frequency of LOH occurred at 9p21-23 band. Their results indicate that more than one tumor suppressor genes at chromosome 9p21-23 region related to a subset of OSCC, while MSI might not be a crucial event. PMID- 12539476 TI - [Evaluation of cariogenic potential of Streptococcus mutans isolated from caries free and -active persons: abilities to synthesize water-soluble and -insoluble glucans]. AB - OBJECTIVE: In this study, authors investigated abilities of Streptococcus mutans (serotype c) strains to synthesize water-soluble and water-insoluble glucans. METHODS: Streptococcus mutans strains were isolated from people with different carious experiences, which were divided into two groups: caries-free (DMFS = 0) group including 19 persons and caries-active group(DMFS > or = 6) including 40 persons. Totally 105 genotypes of Streptococcus mutans strains were obtained, including 22 strains from the caries-free group and 83 strains from the caries active group. The differences of abilities to synthesize water-soluble and water insoluble glucans between these two groups were compared in order to find the correlationship between the synthesis of glucans and caries experience. Then, Streptococcus mutans were cultured in TPY liquid medium containing 1% sucrose in an anaerobic incubator at 37 degrees C for 24 hours. Glucans synthesized by Streptococcus mutans was qualified by using infrared spectrophotometry. The amounts of water-soluble and water-insoluble glucans were measured by using the anthron method. RESULTS: According to the same absorption tops position analysed by infrared spectrophotometry, glucans samples were certified. The amounts of glucans synthesized by Streptococcus mutans strains were different between strains of different genotypes even isolated from the same person. Mostly, the amount of water-insoluble glucans was higher than that of water-soluble glucans between different genotype Streptococcus mutans strains. The oral environments of patients in the caries-active group harbored more Streptococcus mutans strains with higher water-soluble glucans producing ability than those of the caries-free group did (P < 0.05), and patients of the caries-active group also harbored more Streptococcus mutans strains with higher water-insoluble glucans producing ability than those of the caries-free group did(P < 0.01). CONCLUSION: 1. Glucosyltranferases of different genotype strains are different in glucans production, and different genotype strains relate to different cariogenic abilities. 2. Glucans was one of cariogenic factors, and water-insoluble glucans was more important than water-soluble glucans in cariogenic ability of Streptococcus mutans. 3. The differences in glucans production of isolated strains might relate to differences in carious experiences. PMID- 12539478 TI - [A study on tongue cancer cell labeling using bromodeoxyuridine (BrdU) incorporation in vitro]. AB - OBJECTIVE: The aim of this study is to evaluate of Bromodeoxyuridine (BrdU) label in the proliferation of the tongue cancer. METHODS: The authors used 80 micrograms/ml BrdU or iododeoxyuridine (IdU) to label three cases of fresh tissue of tongue cancer and four bottles of Tca8113 cells in logarithmic growth period in vitro. They were fixed and embedded in the routine procedure, and then in serial sections. These specimens were labeled with BrdU polyclonal antibody (1:200), and PCNA monoclonal antibody (1:100), stained using immunohistochemical ABC method. The authors observed the shapes and sizes of those positive cells, counted their label indexes (LI) of BrdU- or IdU-positive cells and PCNA- positive cells respectively. Four bottles of Tca8113 cells were cultured for 1-4 days. One bottle of the cells was every day used in the experimental study about comparing the cells labeled BrdU with those of the S phase of FCM in same cultural time. RESULTS: The positive cells generally owned a larger round nucleus, sometimes in company with sister cells. There was no difference of positive cells in these two kinds of labeling methods between the incorporation of BrdU or IdU and the expression of proliferating cell nuclear antigen (P > 0.05). The cells of S phase with FCM method were much more than that method with BrdU incorporation (P < 0.001). CONCLUSION: The method of BrdU labeling could be used in tumor cellular proliferation studies. PMID- 12539479 TI - [Phospholipase activities of Candida albicans isolates from oral pre-malignant lesions and oral cancers]. AB - OBJECTIVE: The aim of this study is to compare activities of phospholipase of Candida albicans isolates from oral cavities of two groups of patients, including oral pre-malignant lesions and oral cancers. METHODS: A total of 55 isolates, including oral pre-malignant lesions (21), oral cancers (15), and normal health control (19), were studied using the precipitate zone assay on the egg yolk agar. RESULTS: No significant difference was found in phospholipase activities between these isolates from normal controls and oral pre-malignant lesions, while the activities of phospholipase of isolates from oral cancers were higher than those of the normal controls. CONCLUSION: Candida albicans may play different roles in different manners in these two closely related diseases. PMID- 12539480 TI - [Surface stress analysis of distal extension removable partial denture retained with two types of semi-precision attachments]. AB - OBJECTIVE: The aims of this study are to analyze the surface stress of the periodontal supporting bone of the bilateral distol extension removable partial denture which is retained by using intra-coronal or extra-coronal semi-precision attachment, and to characterize the biomechanics of these two designs by using a strain gauge. METHODS: A fresh human mandible specimen with 76|67 missing and six bilateral partial removable denture retained with six semi-precision attachments were made, including three attachments with intra-coronal studs and three attachments with extra-coronal vertical bars. A total of six 45 degrees rosette strain gauges were bonded at six prepared points on the surface of the mandible to measure the surface stress, including the middle point between 76 of the buccal edentulous alveolar crest area under the denture base verge(point 1), the distal buccal cervix of 5 (point 2), the buccal apical area of 5 (point 3), the buccal middle area of 34 roots (point 4, 5), the lingual middle area of 5 root (point 6). The static loads of 14N, 28N and 42N were applied vertically, buccally 45 degrees, lingually 45 degrees at the middle point of the 6|6 occlusal surface. The micro-strain was recorded, and the maximal/minimal principle stresses were calculated for each RPD and each point. The stress characteristics of these two types of attachments were compared and analyzed. RESULTS: In most situations, the stress of these two semi-precision attachments showed significant differences. The vertical load: The stress values of these intra-coronal and extra-coronal attachments at points 1, 2, 3, 4, 6 differed significantly(P < 0.05), including points 1, 2, (sigma intra < sigma extra) and points 3, 4, 6(sigma intra > sigma extra). Buccal loads: The stress values of these intra-coronal and extra-coronal attachments at points 1, 2, 3, 4, 5 differed significantly(P < 0.05), including points 1, 2(sigma intra < sigma extra) and points 3, 4, 5(sigma intra > sigma extra). Lingual loads: The stress values of these intra-coronal and extra-coronal attachments at points 2, 3, 4, 6 differed significantly (P < 0.05, sigma intra > sigma extra). The intra-coronal attachment generally produced higher stress at the site of the alveolar bone around the abutment than the extra-coronal attachment, but extra-coronal attachment produced higher stress at the edentulous alveolar crest and the distal cervical alveolar bone of the distal abutment than the intra-coronal attachment. CONCLUSION: The intra-coronal attachment is suggested to be applied in some cases that the periodontal condition of distal abutment was good, and the extra-coronal attachment is suggested to be used in some cases that the condition of the edentulous alveolar crest is fairly good, while the periodontal condition of the distal abutment was relatively weak. PMID- 12539481 TI - [Establishment and identification of squamous cell lines carcinoma of tongue transferred with anti-sense cyclin A gene]. AB - OBJECTIVE: The aim of this study is to investigate the relationship between cyclin A and cycle regulation of squamous cells carcinoma of tongue, and to provide basis for cancer gene therapy. METHODS: Eukaryocyte expression vector (pAS-A) containing anti-sense and the full-length human cyclin A complementary DNA (cDNA) (1.77 kb) was constructed and was transferred into squamous carcinoma of the tongue cell line (Tca8113) by Lipofect AMINETM introduction. The positive cell clones were selected with G418. Transcription of Neo gene mRNA and cyclin A mRNA were determined by in situ hybridization. The stable expression of anti sense cyclin A in the Tca8113 cell line was determined using immunohistochemical methods. RESULTS: After G418 selection, cells transferred anti-sense cyclin A were obtained successfully. The positive cells of in situ hybridization of specific-stained Neoprobe were observed in cells transferred eukaryocyte expression vector. The positive cells of in situ hybridization of cyclin A in cells transferred anti-sense cyclin A were significantly fewer than those in cells transferred cyclin A. The positive rate of cyclin A immunohistochemical stain in cells transferred anti-sense cyclin A was significantly lower than that in cells transferred cyclin A. CONCLUSION: Human anti-sense cyclin A gene is stably expressed in the Tca8113 cell lines. PMID- 12539482 TI - [Effects of low energy laser on tooth movement and remodeling of alveolar bone in rabbits]. AB - OBJECTIVE: The aim of this study was to investigate effects of low energy laser on the experimental tooth movement and the remodeling of alveolar bone in rabbits. METHODS: A total of 42 white rabbits were chosen and randomized into one control group and six experimental groups, with 6 rabbits in each group. After anesthesia with 2% pentobaritone sodium, orthodontic appliances consisting of a coil spring connected bilaterally the upper first molar with the upper incisor by using a ligature wire. The force exerted at the time of insertion was approximately 80 g. The left side served as the control side, and the right side was the experimental side treated by receiving irradiation of low energy laser. The treatment periods of different groups lasted separately for 1, 3, 5, 7, 14, 21 days respectively. The displacement extent of teeth was measured by employing the computer image analyzing system. The results were analyzed statistically. Through HE staining, the histomorphological character of tissue around first molar was also investigated, and numbers of osteoclasts were counted. RESULTS: The displacement extent of teeth on the experimental side, which was irradiated by low energy laser, was more obvious than that of the normal control side. The difference was statistically significant 1, 3, 14, 21 days after the beginning of the treatment. Through histological observation under a light microscope, the osteoclasts and osteoblasts on the experimental side remained more active than those of the control side. There is significant difference in amount of osteoclasts between the experimental and the control sides, 3, 5 or 7 days after the treatment (P < 0.01, P < 0.05). CONCLUSION: The irradiation of low energy laser promote tooth movement and remodeling of alveolar bone. PMID- 12539483 TI - [A study of analyzing short-chain fat acid by gradient reversed-phase high performance liquid chromatography]. AB - OBJECTIVE: The objective of this study is to obtain the optimized separating conditions of formic acid, acetic acid, propionic acid, butyric acid, valeric acid, lactic acid, succinic acid from bacteria using reversed-phase high performance liquid chromatograph (RP-HPLC), and to set up a reliable progress in identifying the end-products of oral bacteria. METHODS: The conditions of separating seven short-chain fat acids were optimized using the method of gradient RP-HPLC. RESULTS: The study showed that the optimal chromatographic parameters were: 1.0 ml/min and 2.0 ml/min of flow rates, and the wavelength of ultraviolet detection was 217 nm. The composition of eluent: the ratio of phosphate buffer (pH 2.8) to methanol is 95:5 (v/v), gradient 5%-50%. The liner gradient wash will be performed in 10 minutes at room temperature. The average recovery rate was 92%, and RSD was 1.8%. CONCLUSION: The results of this paper will be helpful to determine acid composition of end products of oral bacteria. PMID- 12539484 TI - [A retrospective clinical study of 6539 cases of malignant oral-maxillofacial tumors]. AB - OBJECTIVE: The aim of this study is to obtain the general information of post operation malignant oral-maxillofacial tumor patients in forty-seven years. METHODS: A total of 6539 patients with malignant oral-maxillofacial tumors in the hospital of stomatology, west china university of medical sciences, from 1953 to 2000 were retrospected. RESULTS: The average age of patients with malignant oral maxilloficial tumor is 48.7 years old. The ratio of male to female is 2.31:1. The most common original malignant tumors in this oral-maxillofacial region are malignant oral tumors, malignant maxillary tumors, malignant mandibular tumors, and malignant sialoma successively. As to the original tissues of tumors, there were 5869 cases of epithelial tumors, and made up 89.75% of all the investigated patients. Tumors came from lymphatic and hematopoietic systems and mesoderm were much less those came from epithelia. There were 4177 cases of squamous cell carcinomas and the percentage was 63.88%. CONCLUSION: The incidence rate of malignant tumors in oral-maxillofacial region tends to increase, and the average age of patients also rose. The ratio of male to female decreases gradually. Epithelial tumors, especially squamous cell carcinomas are very common tumors happening in this region. PMID- 12539485 TI - [Nutritional status assessment of patients with oral maxillofacial cancers]. AB - OBJECTIVE: The objective of the study is to evaluate the nutritional status of patients with oral maxillofacial cancer. METHODS: The nutritional status was evaluated using seven easily determined variables and multi-factor nutritional index in 50 hospitalized patients with oral maxillofacial cancer from 1990 to 1992. RESULTS: Fifty-six percent of patients had protein-energy malnutrition. The high incidence of protein-energy malnutrition was found in those patients without adequate dietary intake. CONCLUSION: Patients with oral maxillofacial cancer usually have protein-energy malnutrition. The reason is that the patients always have an inadequate dietary intake. PMID- 12539486 TI - [Expression of vascular endothelial growth factor in oral squamous cell carcinoma]. AB - OBJECTIVE: The aim of this study was to estimate the expression of vascular endothelial growth factor(VEGF) and intratumoral microvessel density (IMVD) in oral squamous cell carcinoma(SCC). METHODS: A total of 60 cases of oral squamous carcinoma were immunohistochemically stained with a SP method by using monoclonal antibodies of VEGF and CD34. RESULTS: There was no significant difference in the expression of VEGF in different TNM stages and lymphe node metastases of oral squamous cell carcinoma. The expression of VEGF showed no relation with pathologic grades, but IMVD expression differed significantly in SCC. The carcinoma in grade I and IV of IMVD differed significantly in expression of VEGF. CONCLUSION: It indicates the change of IMVD may resulted from the change of VEGF in oral SCC. PMID- 12539487 TI - [Anatomical and functional studies on surgical and non-surgical treatment of mandibular condylar process fractures]. AB - OBJECTIVE: This study aim at comparing anatomical and functional recovery from surgical and non-surgical treatment of condylar process fractures. METHODS: Totally Forty-nine patients, 22 treated by performing operation, and 27 using non surgical methods, were included in this study. Displaced angle of condylar, maximal mouth opening, lateral deviation on opening and masticatory function were analyzed using radiological and clinical examinations 6 weeks and 1 year after treatment. RESULTS: Patients treated by surgical method had significantly greater initial displaced angle of condyles than did the group treated non-surgically, but patients treated non-surgically had significantly greater displaced angle of condylar and lateral deviation on opening than did the group treated surgically 6 weeks and 1 year after the therapy finished. Patients treated surgically had significantly greater masticatory function than did the group treated non surgically 6 weeks and 1 year after treatment. Six weeks later, patients treated surgically had significantly greater maximal mouth opening than did the group treated non-surgically. CONCLUSION: Surgical treatment may be helpful in anatomical and functional recovery of mandibular condyles for patients with displaced condylar process fractures. PMID- 12539488 TI - [A comparative study of clinical application of two types appliances for maxillary molar distalization]. AB - OBJECTIVE: The aim of this study is to evaluate advantages and disadvantages of two types of appliances for maxillary molar distalization. METHODS: Totally, 28 patients with class II malocclusion resulting from molars shifting mesially were treated using two different molar distalization methods, including using removable and fixed appliances. An intra-oral removable acrylic appliance with headgear for extra-oral traction resulted in tipping the maxillary first molars distally. The modified rapid helix expansion appliance (MRHEA), which consisted of bands, anchorage teeth, Nance palatine bases, helix bars and connecting rods, made the maxillary first molars move parallel distally. RESULTS: The desired efforts were gained in 7 or 9 months by using removable appliances, but only 2 or 2.5 months if using MRHEA. The treatment results of using removable appliances were dependent on patients' cooperation, however, the method of using fixed appliance overcame some shortcomings of other molar distalization techniques, and this appliance would not lead to the distal inclination of the crown of the shifted teeth. CONCLUSION: These two methods could convert the class II molar relationship to class I. The MRHEA can save much chair-side time, and shorten the period of treatment, move molars distally parallel. PMID- 12539489 TI - [Effects of para-aminobenzoic acid (PABA) on growth of Streptococcus mutans]. AB - OBJECTIVE: The aim of this study is to examine effects of para-aminobenzoic acid (PABA) on the growth of Streptococcus mutans (S. mutans). METHODS: Different concentrations of PABA (10(-10)-10(-3) g/L) were separately transferred to modified Carlsson medium. S. mutans (ATCC 25175) grew in modified Carlsson medium with different concentrations of PABA. All cultures were incubated at 37 degrees C anaerobically in an atmosphere of 80% of nitrogen (v/v), 10% of hydrogen (v/v) and 10% of carbon dioxide (v/v) for 48 hours. The absorbance values of S. mutans were measured by using a spectrometer (UV-1601). The colony forming units (CFU) were obtained by growing S. mutans in media with different concentrations of PABA (10(-10)-10(-3) g/L). RESULTS: Different concentrations of PABA had different stimulating effects on the growth of S. mutans (P < 0.05). But this kind of stimulating effects declined when the concentration of PABA was 10(-3) g/L. CONCLUSION: This experiment indicates PABA has stimulating effects on the growth of S. mutans, and PABA can promote growth of S. mutans. PMID- 12539490 TI - [Effects of para-aminobenzoic acid (PABA) on growth of Lactobacillus acidophilus]. AB - OBJECTIVE: The aim of this study is to examine effects of para-aminobenzoic acid (PABA) on the growth of Lactobacillus acidophilus (L. acidophilus). METHODS: Different concentrations of PABA (10(-10)-10(-3) g/L) were separately transferred to the modified Carlsson medium. L. acidophilus (ATCC4356) grew in these Carlsson media. All cultures were incubated at 37 degrees C anaerobically in atmosphere of 80% of nitrogen, 10% of hydrogen, and 10% of carbon dioxide for 48 hours. Absorbance values (lambda = 540 nm) of bacterial suspensions were measured using a spectrometer (UV-1601). Colony forming units (CFU) were obtained by growing L. acidophilus in Carlsson media with different concentration of PABA (10(-10)-10( 3) g/L). RESULTS: Different concentrations of PABA (10(-10)-10(-4) g/L) had different stimulating effects on the growth of L. acidophilus (P < 0.05). But stimulating effects declined, when PABA concentration was 10(-5) g/L, and when the concentration of PABA reached 10(-3) g/L, the stimulating effect disappeared. CONCLUSION: This study indicates PABA stimulates the growth of L. acidophilus, and PABA can promote growth of L. acidophilus. PMID- 12539491 TI - [IL-1 beta mRNA and TNF-alpha mRNA expression in gingival tissues of patients with adult periodontitis]. AB - OBJECTIVE: The aim of this study was to examine interleukin-1 beta (IL-1 beta) mRNA and tumor necrosis factor alpha (TNF-alpha) mRNA expression in gingival tissues of patients with adult periodontitis. METHODS: A total of 19 patients, including 10 patients with adult periodontitis and 9 healthy individuals as the control, were evaluated for were selected for evaluating IL-1 beta mRNA and TNF alpha mRNA expression, using reverse transcription polymerase chain reaction (RT PCR) technique. RESULTS: The relative amount of IL-1 beta mRNA expression of gingival samples from patients with adult periodontitis (0.819 +/- 0.045) (P < 0.01) was much higher than that from the normal control (0.306 +/- 0.087) (P < 0.01). The relative amount of TNF-alpha mRNA expression, gingival samples from AP patients (0.696 +/- 0.098) was also much higher than that from the normal control (0) (P < 0.01). CONCLUSION: As the inflammatory and bone absorption mediators, IL 1 beta and TNF-alpha might play important roles in the pathogenesis and development of adult periodontitis. PMID- 12539492 TI - [Changes of osteocalcin in serum of young SD rats after functional protrusion]. AB - OBJECTIVE: The aim of this study is to investigate temporal and spatial change patterns of osteocalcin in serum of young growing SD rats, in conditions of different functional treatment time spans. METHODS: Totally, 72 SD rats are randomly divided into 3 groups, including the control group, the 24-hour wearing group (wearing functional appliance 24 hours a day), and the 12-hour wearing group (wearing appliance for 12 hours in daylight). One week after the experiment, rats were sacrificed and osteocalcin level in serum was measured by using radioimmunoassay. Date was processed using macroscopic and microscopic analysis. RESULTS: Serum osteocalcin had circadian rhythm and the peak was about 12:00 in a day. After wearing the functional appliance, serum osteocalcin level increased with treatment time, and serum osteocalcin level in the 24-hour wearing group rose more obviously than that in the 12-hour wearing group. CONCLUSION: For the functional appliance, better results can be achieved in the 24-hour wearing group. PMID- 12539493 TI - [A preliminary study on relationship between concentrations of human growth hormone in urine (HGH-U) and puberty]. AB - OBJECTIVE: The aim of this study is to investigate relationship between the concentration of human growth hormone in urine (HGH-U) and puberty, and to find a simple, accurate, and acceptable way of judging puberty of patients. METHODS: The concentration of HGH-U was measured by using radioimmunoassay in 277 students of Chengdu, and students were divided into three different groups, according to their ages (9, 13, 17 years old). Half of them are male. In order to evaluate the relationship between the concentration of HGH-U and puberty, the information of age, height, weight, growth velocity and sex character of all individuals were obtained, and analyzed their relation with the concentration of individuals' HGH U. RESULTS: 1. There was no significant difference in the concentration of HGH-U between the 9-year-old group and the 13-year-old group, but there is significant difference between the concentration of HGH-U in the 9-year-old, 13-year-old groups and that of the 17-year-old group, and the concentration in the latter group is lower than those of the former two groups. 2. The concentration of HGH-U was negatively correlated with height, body weight, age, sex character and growth velocity (P < 0.05), but the correlation coefficient is low. 3. The coefficients of variation of HGH-U in all group were higher than 30%, which indicated a low dispersion degree. CONCLUSION: The concentration of HGH-U cannot be used as an independent indicator to evaluate puberty of children. PMID- 12539494 TI - [A study on the relationship between deciduous tooth caries in pre-school children and their neonatal conditions]. AB - OBJECTIVE: This study was designed to examine whether the neonatal conditIon might be related to the caries of deciduous teeth in pre-school children. METHODS: 1000 children from 3 years old to 6 years old in Chengdu were surveyed, and 260 children with dmft > or = 6 and 260 caries-free children were chosen. Data was collected by 200 pairs of useful questionnaire from their parents and analyzed by using logistic regression analysis and chi 2 test. RESULTS: The neonatal condition was closely related with incidence rates of deciduous caries in pre-school children (P < 0.05). The incidence rate of children with premature delivery was higher than that with full-term normal delivery (P < 0.05). No significant difference was showed in children with full-term normal delivery and children with dystrophy (P < 0.05). CONCLUSION: There should be emphasis on the children with premature delivery in caries preventive cares. PMID- 12539523 TI - [Comparative histomorphological study on the craniomaxillary development of the trisomy-18 with cleft palate and euploid mice]. AB - OBJECTIVE: The aim of this study is to compare histological morphology of craniomaxillary complex of Trisomy 18 with cleft palate (Ts + CP) and euploid mouse foetuses to understand whether cleft palate is associated with maldevelopment of the craniomaxillary complex or with hypoplasia of the adjacent tissues. METHODS: Thirty pairs of Ts + CP and euploid mouse foetuses with about equal body weight were selected, and sixty craniomaxillary complex were precisely orientated in the coronal plane, then serially sectioned with 7 microns thickness through the primary palate, the whole hard palate and the soft palate. Finally six sections were selected for comparative histomorphological study based on anatomical landmarks. RESULTS: The trisomy with cleft palate mouse foetuses had significantly developmental hypoplasia in the primary palatal shelves. The palatal shelves, vomer, palatal bone and nasal septal cartilage in cleft samples showed significantly developmental hypoplasia with a relatively wider nasal septum, nasal capsule and a bigger nasal airway volume, nasopharyngeal cavity. CONCLUSION: The hard tissues of the craniomaxillary complex in Ts + CP mouse foetuses, including bone and cartilage show serious maldevelopment or developmental hypoplasia, and these may be closely related to the autosomal trisomic condition. PMID- 12539524 TI - [Histological study on the polyhydroxybutyric ester(PHB) membrane used for guided bone regeneration around titanium dental implants]. AB - OBJECTIVE: To evaluate the effects of PHB membrane around dental titanium implants used in guided bone regeneration technique. METHODS: Endosseous cylindrical implants were placed in dogs' mandibles after extraction of all the mandibular premolars and the first molars immediately. Ideal implant placement sites were modified by creating standardized 3 mm x 5 mm facial dehiscent defects. The defects were covered with PHB membrane and titanium membrane respectively, then the membranes were fixed on the periosteum by sutures, and the remainder served as controls. Tissue closure was achieved with mattress and interrupted single sutures, and antibiotic injection was given to the dogs in the following 3 days. The dogs were divided into three groups according to the healing time of 1, 2 and 3 months, then the three groups of dogs were sacrificed. After that, tissue blocks containing the implants and the control ones were removed and studied by gross observation, X-ray radiograph and fluorescence microscope, and the surfaces of the membranes were observed by scanning electron microscope. The specimen used for light microscope study were fixed with 10% formaldehyde for 24 hours, and decalcified with 0.5 mol/L EDTA for 2 weeks. After the titanium implants were carefully removed, the tissue blocks were embedded into the paraffin wax, and the sections were obtained. RESULTS: The PHB membrane degraded slowly and partly 3 months after placement, and the biodegradable property of the PHB membrane deserved a longer period of study; The bone defects covered with PHB and titanium membrane were filled with much more new bone than that of the control group. CONCLUSION: PHB membrane can enhance bone formation and can be used for guided bone regeneration. PMID- 12539525 TI - [A study of the mechanism of effects of solution containing trace elements on remineralization layer formation of enamel carious lesions]. AB - OBJECTIVE: To study the effects of several remineralization solutions with different composition on the artificial demineralization lesions of bovine enamel and the mechanism of remineralization layer formation. METHODS: 36 bovine teeth were selected and artificial carious lesions were formed with lactic acid gel. Then all specimens were divided into 4 groups which contained no fluoride, fluoride, fluoride-calcium-phosphate-tin, and de-ionic water separately, and the de-ionic group was taken as control. The structure and composition of the sediments on the enamel surface were evaluated and detected by X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM). Histological change of enamel section was examined by polarized microscopy. RESULTS: Compared with the control group, all the solutions had remineralization effects, especially the solutions containing trace elements. XPS showed that there were sediments formed on the surface of carious lesion, the compositions of the sediments were CaF2, HAP and non-specifically absorbed fluoride (NSAF). Irregular, crystal deposits were also found under SEM on the enamel surface. The polarized microscopy results showed that the observed demineralization areas decreased after exposure to the solutions. CONCLUSION: Solutions containing fluoride and trace elements were effective on remineralization. The possible mechanism is that CaF2 and NSAF, instead of FA formed in the remineralization layer. Moreover, the remineralization effects of the fluoride solutions containing trace elements are more outstanding than that without trace elements. The potential mechanism is the synergistic action of fluoride and trace elements. PMID- 12539526 TI - [Ultrastructure of the lymph capillaries in human dental pulp]. AB - OBJECTIVE: The purpose of this article is to observe the ultrastructures of the lymph capillaries in the human dental pulps. METHODS: Investigations have been performed on dental pulps from 8 human permanent teeth, including 4 mandibular first premolars and 4 maxillary first premolars, which were extracted for orthodontic reason. As soon as possible, we cleaved the teeth longitudinally and immersed in the fixative solution at 4 degrees C for 1 hour, then kindly removed the pulps and immersed in the same fixative solution for 3 hours, then routinely postfixed, dehydrated, embedded, and prepared the ultrathin sections (50-70 nm) without semithin section localization. The lymph vessels were distinguished from blood vessels on ultrathin sections by their morphological features. RESULTS: We have observed three fundamental kinds of intercellular junctions, overlapping(51%), end to end(36.1%), and interdigistating (10.6%). A few complex junctions also could be seen, but we failed to find the open junction. The Weibel Palade bodies were occasionally present in the lymphatic vessels endothelial cells of young dental pulps. And we even found the lipofuscins in the cytoplasm of the endothelium. Some inclusion bodies, such as paracrystalline inclusions and myeloid bodies, also could be seen. There were many micropinocytotic vesicles in the cytoplasm of the lymphatic endothelium, the diameter of the most vesicles was about 50-80 nm, some was about 100-500 nm. Some vesicles appeared to lie free within the cytoplasm, and some were seen to touch or open onto the luminal or abluminal surfaces of the endothelia, which suggested that many vesicles were not simple discrete units but rather from parts of chains that reached either luminal or abluminal surfaces. It was reported that micropinocytotic vesicles, open junctions and intraparietal channels provided fluid transport. CONCLUSION: Dental pulp is a special organ, and the lymph capillaries in it have their own ultrastructural features. PMID- 12539527 TI - [Influences of porcine enamel matrix proteins on MC3T3-E1 osteoblast proliferation and differentiation]. AB - OBJECTIVE: Some research suggested that enamel matrix proteins(EMPs) not only induced formation of cementum, but also could promote regeneration of periodontal tissue and alveolar bone. In order to recognize the mechanisms of periodontal regeneration induced by EMPs and provide a basic data for further study, influences of EMPs on osteoblast proliferation and differentiation are explored in this study. METHODS: Porcine enamel matrix proteins were extracted by using acetic acid. MC3T3-E1 osteoblasts were selected and cell culture was used in vitro. EMPs with different concentrations of 50 micrograms/ml, 100 micrograms/ml and 150 micrograms/ml were added separately in alpha-MEM culture medium and compared with negative group and positive group (TGF-beta 1). The count of cell was recorded after 0, 1, 2, 3, 4, 5, 6 days, then the data were analysed statistically by using SAS. RESULTS: The cell count of all EMPs groups were higher than that in the negative control group (P < 0.01), 100 micrograms/ml as well as 150 micrograms/ml EMPs groups were higher than the positive group(TGF beta 1) on the second day (P < 0.01). The cell count of 100 micrograms/ml EMPs group was not only higher than C and T groups, but also higher than that of other EMPs dose groups on the fifth day. CONCLUSION: The proliferation and differentiation of osteoblasts can be promoted distinctly by enamel matrix proteins. PMID- 12539528 TI - [A three-dimensional finite element analysis of the correlation between lengths and diameters of the implants of fixed bridges with proper stress distribution]. AB - OBJECTIVE: The study is designed to investigate the correlation between lengths and diameters of the implants of fixed bridges with proper stress distribution. Meanwhile, the correlation between the diameter of the implants and the size of a mandible bone is evaluated by stress analysis. METHODS: According to the stress around the implants of an implant fixed bridge with proper stress distribution, especially peak stress, a three-dimensional finite element method was used to determine the correlation between diameters and lengths of implants with their diameters or lengths changing. RESULTS: The peak stress surrounding the implants in fixed bridge supported by clinical implants that were commonly used was figured out, and the correlated amount of diameters or lengths of the implants were achieved. Furthermore, a correlation curve of diameter and length of the implants was made. But the correlated amount of diameters or lengths of the implants were not found in this model. CONCLUSION: The proper stress distribution of implants with different implant sizes can be achieved by adjusting diameters or/and lengths of implants. The selection of the implant size is related to the limited model size. Being a preliminary theory, the results will be proved to be clinically acceptable to the size selection of implants. PMID- 12539529 TI - [Clinical appliance of magnetic attachments in mandibular complete overdentures]. AB - OBJECTIVE: To observe the clinical effect of magnetic attachment with overdenture. METHODS: Ten subjects were selected and treated with Magnetic EX-600 retained mandibular complete overdentures. Each patient's masticatory function, overdenture retention force and abutment condition were measured at different periods and the statistics of each individual were compared respectively. RESULTS: Magnetic attachments can provide good retention for mandibular complete overdentures and improve the masticatory performance of the patients. What's more, the mobility and periodontal index of abutments were improved after 6 months. CONCLUSION: Magnetic EX-600 is very useful to improve the function of mandibular overdentures. PMID- 12539530 TI - [Distribution of Prevotella intermedia in the periodontal plaques of children and their parents]. AB - OBJECTIVE: To investigate the distribution of Prevotella intermedia (Pi) among family members by culturing technique and biochemical identification. METHODS: Of all 60 families, 181 subjects were included in the study. Supragingival and subgingival plaques were collected and cultured, 279 strains of black-pigmented gram-negative anaerobes were obtained. Biochemical identifications were performed. RESULTS: Pi could be identified from parents and children with healthy periodontal status. The prevalence of Pi in children was 70.49%, which was significantly higher than that in adults (43.33%). CONCLUSION: The result indicates that Pi might be an opportunistic periodontal pathogen that could be colonized in periodontal plaques easily and early. To study the exact role of Pi in the pathogenesis of periodontitis, it is necessary to do longitudinal following study of the subjects. PMID- 12539531 TI - [Expressions of five protein kinase C isoforms in salivary adenoid cystic carcinoma]. AB - OBJECTIVE: The protein kinase C (PKC) family of isoforms plays the key role in the regulation of cellular events, including cell proliferation, differentiation and canceration. Previous work from our laboratory has shown the alteration of PKC and its inhibitor activities in salivary adenoid cystic carcinoma (SACC) [Sun Changfu, et al. West China Journal of Stomatology, 1998, 16(3): 201-203]. This study aimed to further explore the relationship between PKC isoforms and canceration of SACC. METHODS: Expressions of five isoforms of PKC (alpha, beta I, beta II, epsilon and delta) were examined in cytosolic and membranous fraction of specimen homogenates from SACCs and were compared with the adjacent normal tissues by immunoblotting. Equal amounts of cytosolic and membrane protein from control and experimental groups were simultaneously subjected to sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE), transferred to nitrocellulose membranes, and reacted with first and second antibodies; Color development and image analysis were performed. RESULTS: 1. Cytosolic PKC-alpha, PKC-beta I, and PKC-beta II were expressed in SACCs and adjacent normal tissues. The Expressions of these three cytosolic isoforms were reduced significantly in SACCs compared with the adjacent normal tissues; 2. Membranous PKC-alpha was only detected in SACCs and the membranous translocation of cytosolic PKC-alpha was also seen. 3. The expressions of PKC-epsilon and PKC-delta were not detected in both SACCs and adjacent normal tissues. CONCLUSION: There was close relationship between changes of expression of PKC isoforms (alpha, beta I, beta II) and carcinogenesis of SACC. We postulated that there may be relationship between the membranous translocation of PKC-alpha and hematogenous lung metastasis in SACC. PMID- 12539532 TI - [Effects of ultrasonography on evaluating the status of cervical lymphatic metastasis in patients with oral squamous cell carcinoma]. AB - OBJECTIVE: To investigate effects of ultrasonography on evaluating the status of cervical lymphatic metastasis in patients with oral squamous cell carcinoma (OSCC). METHODS: Nineteen OSCC patients with the diagnosis confirmed by biopsy of primary focus were included in this study. All the patients were hospitalized in the department of stomatology, Beijing Medical University during June, 1995 and June, 1996, among them there were 13 men and 6 women. The ages were ranged from 31 to 71 years old, with the average of 54.4 years old. The primary focus of 13 cases were in tongue, 3 cases in cheek, 2 cases in gum, and 1 case in the conjunction of gum, mouth floor and tongue respectively. All the patients were examined by ultrasonography prior to operation, and operated with the procedures of excision of primary focus and radical neck dissection. The length, width, Vmax and Vmin and Resistance Index (RI = (Vmax-Vmin)/Vmax) of blood flow of cervical lymphatic nodes were measured with acuson 128 xp/10 color Doppler flow imaging scanner with a 7.5 MHz probe. The total of 34 cervical lymphatic nodes were examined by the scanner. L/W ratios in all nodes, RI and Vmax in 16 nodes were obtained respectively. All results were divided into two groups, metastases and non-metastases groups based on the pathological findings for further statistic analysis. Student t test was employed in the study. RESULTS: The average L/W ratios, the average Vmax and the average RI were 1.95 and 2.17, 0.24 and 0.14, and 0.80 and 0.56 in metastases and non-metastases groups respectively. There was a significant difference between RI in two groups, but no statistical differences were revealed in L/W ratios and Vmax, despite the great difference in the average Vmax in the two groups. When RI > or = 0.65 and Vmax > or = 0.20 m/s were regarded as the criterion for positive diagnosis, the accuracy rate, the sensibility and the specificity was 91.2%, 90%, and 91.7% respectively in making diagnosis of OSCC. CONCLUSION: The status of cervical lymphatic metastases in OSCC patients could be evaluated accurately by the color Doppler flow imaging scanner, which might be helpful in making accurately diagnosis of OSCC, selecting the appropriate operation procedure of neck dissection leading to increased survival ratio and improved quality of life for the patients. PMID- 12539533 TI - [Oncocytic carcinoma of salivary gland]. AB - OBJECTIVE: To study the histological and immunohistochemical characteristics of oncocytic carcinoma of salivary gland. METHODS: 7 cases of oncocytic carcinoma of salivary gland were investigated by HE, PAS, PTAH staining and immunohistochemistry. RESULTS: Among the cases, 4 were male and 3 were female. The average age was 49 years old. 3 cases occurred in the parotid, 1 cases in submandibular, 1 cases in sublingual and 2 cases in minor salivary gland of palate. Investigated histologically, the tumour cells are characterized by oncocyte. The evidence of cellular atypia, obviously increased mitoses and infiltrative growth pattern are indicative of malignancy. Immunohistochemically, alpha 1-ACT, alpha 1-AT, LF, TF and CEA were all strongly positive in the normal salivary cells and adenolymphoma, while in oncocytic carcinoma, alpha 1-ACT and alpha 1-AT were moderate positive and LF, TF and CEA were light positive. CONCLUSION: 7 cases were characterized by oncocyte and showing infiltrative growth, alpha 1-ACT, alpha 1-AT, LF, TF, CEA were positive in oncocyte. PMID- 12539534 TI - [Three-dimensional surface reconstruction of spiral CT for teeth and clinical use in examining impacted teeth of jaws]. AB - OBJECTIVE: To establish a new technique for three-dimensional surface reconstruction of teeth (dental surface reconstruction), and apply it to investigate impacted teeth in bone before operation. METHODS: On the basis of three-dimensional surface reconstruction of teeth and jaws, the dental surface reconstruction is obtained with the technique of shaded surface display (SSD), making transparent bone around. This technique is applied to investigate 9 cases of supernumerary and impacted teeth. RESULTS: Dental surface reconstruction image can present the dental surface image including crown, root, neck and root bifurcation in three-dimensional way, without presenting the image of other tissue around. In 4 cases, the abnormal configuration, labial or palatal location, and relation with dentition in anterior maxilla of 5 supernumerary were shown. In 1 case, the shape, location, eruption orientation and the angle of orthodontic distraction of 1 impacted permanent canine were displayed. In 3 cases, the shape, location, and extraction resistance of 6 impacted wisdom teeth were revealed. In 1 case, the configuration, distance to adjacent molar of 1 impacted wisdom tooth, which is a transposition tooth in maxillary sinus, were also demonstrated. CONCLUSION: The dental surface reconstruction is a new technique to display three-dimensional surface configuration of teeth. This technique can be applied to examine impacted and supernumerary teeth before surgical and orthodontic treatment. PMID- 12539535 TI - [A study on the clinical classification of oral submucous fibrosis]. AB - OBJECTIVE: To increase the positive rate and sensitivity of the diagnosis for oral submucous fibrosis (OSF). METHODS: A general oral survey of 11,046 persons was carried out in Xiangtan, Hunan. According to Pindborg's diagnostic standard to epidemiological studies of OSF, 335 cases of OSF were found. The prevalence was 3%. According to different clinical manifestations, 335 cases of OSF were divided into two types: diffuse type and localized type. The cases with typical features of OSF, such as burning pain and discomfort, dryness of the mucosa, progressive inability to open the mouth and presence of palpable bands in the oral mucosa and so on, were in the diffuse type. Those with non-typical features, only having presence of palpable bands in the oral mucosa, were in the localized type. 72 cases were followed-up for ten years. RESULTS: 335 cases of OSF were divided into two types. 13 cases (3.9%) were in the diffuse type, and 322 cases (96.1%) were in the localized type. Most of the OSF in this region belonged to the localized type. It was also discovered that without proper treatment, the subjects sticked to one type of OSF without changing from one type to another after ten years of follow-up. CONCLUSION: Localized type of OSF is the major type of OSF in Xiangtan. It is an independent clinical type. To classify OSF, it is good for the clinicians to pay attention to non-typical OSF. If the non-typical OSF cases are taken into consideration, the prevalence of OSF will increase considerably. PMID- 12539537 TI - [The diel rhythm of PGE2 in blood of the growing rat after retractive force on mandible]. AB - OBJECTIVE: To study whether the orthopedic therapy will affect the rat's metabolism, the author investigates the diel rhythm of PGE2 in blood of the growing rats before and after retractive force on mandible. METHODS: 72 rats which were 4 weeks old and about 90 g, were divided equally and randomly into three groups: the contrast group were not exerted force, the daytime-forced group were exerted force from 8:00 am to 20:00 pm, the nighttime-forced group were exerted force from 20:00 pm to 8:00 am of the next morning. The backwards force imposed on the rats were exerted through the center of the mandibular condyle and was about 40 g(0.4 N). The period of experimentation was 7 days. In the 8th day, every 4 rats from each of the group were killed respectively at 0:00, 4:00, 8:00, 12:00, 16:00 and 20:00 to collect 3 ml blood. Then radioimmunoassay methods were used to measure the concentration of PGE2 in the blood. RESULTS: In natural state, there was a diel rhythm in the concentration of PGE2 in rats' blood and the diel rhythm was related to that of endogenous PGE2 in condyle; After imposed orthopedic force to restrain mandibular growth, the normal diel rhythm of PGE2 in blood disappeared. There was no significant difference in the time-concentration curves between the contrast group and the daytime-forced group (P > 0.05), but the difference between the contrast group and the nighttime-forced group was obvious (P < 0.05) and the mesor of the nighttime-forced group was greatly decreased. CONCLUSION: The concentration of PGE2 in rats' blood will not be affected by the orthopedic force in daytime, in nighttime. For this reason, not only the therapeutic effect but also the effect on the body's metabolism should be considerd in clinical therapy, and further researches should be carried out on whether the local orthopedic force would influence the metabolism of the whole body. PMID- 12539536 TI - [Circadian rhythm changes of the levels of endogenous insulin in growing rat condylar cartilage after functional mandibular protrusion]. AB - OBJECTIVE: In order to determine which time of the day is best to wear functional appliances in clinic, the author investigates the circadian rhythm of the level of endogenous Insulin in growing rat condylar cartilages and the changes after functional mandibular protrusion in daytime (8:00-20:00) or nighttime (20:00 8:00). METHODS: 72 4-week-old SD rats were divided equally and randomly into experimental and control groups, and the rats in experimental groups were put on the functional appliance in daytime (8:00-20:00) or nighttime (20:00-8:00). After 7 days, all rats were sacrificed at 0:00, 4:00, 8:00, 12:00, 16:00, 20:00 in a day. Radioimmunoassay methods were used to detect the levels of endogenous insulin in growing rat condylar cartilages. RESULTS: 1. The levels of endogenous insulin in the growing rat condylar cartilage displayed a significant circadian rhythm. The acrophase was about 12:13. 2. The circadian rhythm still existed after the rats had been put on the functional appliance. Both mesor and amplitude were significantly higher in the experimental groups than in the control group 3. The levels (both mesor and amplitude) of endogenous insulin in the condylar cartilage were significantly higher in the daytime-forced group than in the nighttime-forced group. CONCLUSION: 1. The growth and remodeling of the condylar cartilage in the growing rats shows significant circadian rhythm. The most active period of chordrocyte proliferation may occur during the daytime. 2. The circadian rhythm still exists after the rats have been put on the functional appliances. 3. Compared with nighttime appliance wearing, daytime appliance wearing may be more conductive to cell proliferation, so it may be more effective to stimulate the growth and remodeling of condylar cartilage in the growing rat. 4. Related to human beings, these data suggest that the best period for patients to wear the functional appliances might be from 20:00 pm to 8:00 am of the next day. PMID- 12539539 TI - [3D finite element analysis of stress distributions in supporting tissues of clasp-type partial dentures of transferring occlusion force]. AB - OBJECTIVE: To provide a theoretical basis for clinical application of the clasp type partial denture of transferring occlusion force, the stress distributions in its supporting tissue were analyzed. METHODS: The 3D finite element model was made, and the features of stress distributions in the supporting tissues of the clasp-type partial dentures which had different rest recess angles were discussed. RESULTS: The medial stress component was significantly larger than the distal one in the supporting tissue around main abutment root. When the rest recess angles were equal or greater to 60 degrees, the stress values were greatly reduced in the supporting tissue around the roots of the three abutments. When the rest recess angle reached 75 degrees, the stress values in the supporting tissue around the main abutment root wore decreased by 30% than the angle 0 degree and by 52% than the angle 45 degrees, the stress values in the supporting tissue around the root of second abutment (adjacent to the main abutment) were wholly increased by 44% than the angle 0 degree and reduced by 23% than the angle 45 degrees, the stress values in supporting tissue around the root of third abutment (outside of the second abutment) were wholly increased by 53% than the angle 0 degree and decreased by 31% than the angle 45 degrees, the stress values in the medial root of the main abutment were reduced by 73%. CONCLUSION: The clasp-type partial denture can resolve part of the occlusion force into two components the medial and distal components, and the occlusion force is distributed to several abutments, at the same time the conversed force components can be canceled by each other. So the whole stress values are greatly reduced and the stress around abutment root is greatly decreased. As a result the damage to the abutment will be eliminated. A design with centralized multi abutments and multi fixed bodies are used in this denture, which are more fixed and stable and have no obvious sinkage in the plate base. Therefore, the mastication function can be satisfactorily recovered, and the reasonable rest recess angle for the clasp-type partial denture of transferring occlusion force is about 75 degrees. PMID- 12539538 TI - [The preliminary analysis on the changes of the oral functional space in Class II division 1 malocclusion after triggered by functional appliance]. AB - OBJECTIVE: The purpose of this paper is to study the area changes of the oral functional space in the patients with Angle's Class II division 1 malocclusion after triggered by FR-I functional appliances. METHODS: Twenty patients with Angle's Class II division 1 malocclusion were selected, and the patients were selected by the standards of an horizontal growth pattern with normal maxilla, retracted mandible and short lower facial height. All patients were triggered by FR-I functional appliances and achieved the Angle's Class I occlusion after treatments. The average periods of the treatments were about 9 months. The assessments were made from the lateral cephalograms taken in centric occlusion before and after treatments. The upper and behind borders of the oral functional space are hard palate and soft palate, the forward border is the lingual side of upper and lower incisors, the lower border is the line between apex of lower incisor and lower first molar. The graphics from the lateral cephalogram were painted and input into the computer by scaner, which were measured by special software. Self-controls of individuals before and after treatments were used in statistic analysis. RESULTS: The paired-samples t test confirmed the oral functional space was enlarged after triggered by FR-I functional appliance, which was induced by obviously increased mandibular body and ramus, anterior lower facial height and posterior facial height. At the same time, the angle of Y axis increased and the upper incisor inclined lingually, but the inclining angle of lower incisor was not obvious. CONCLUSION: Enlarged oral functional space facilitates the development of the maxilla, mandible and alveolar bones and consummates the functions of oral and jaw system. This study aids the design and application of FR-I functional appliance. PMID- 12539540 TI - [Effects of maxillary growth of Wistar rats with bilateral artificial cleft palate after premaxillary orthopedic treatment on expression of proliferating cell nuclear antigen(PCNA)]. AB - OBJECTIVE: Being based on the principle of Latham's appliance, this experiment is designed to detect activity of osteoblasts in the maxillary sutures of Wistar rats with bilateral artificial cleft palate by immunohistochemistry technique, and evaluate the effects of this appliance on the growth and development of the maxilla. METHODS: The animal models of male infant Wistar rats with bilateral cleft palate were made by removing some palatal bones, splitting a "V" gap of 1.5 cm wide at the line between premaxilla and segments of maxilla. A sort of appliances, which could be fixed in the mouth of Wistar rats with bilateral cleft palate for correcting protrusion premaxilla was constructed basing on the principle of Latham's appliance. Then the diferent pathological changes of osteoblast proliferation between the experimental group and the two controlled groups were examined. The jugomaxillary sutrues, temporomalar sutures and sphenoipalatine suture were harvested 7, 14 days after premaxillar orthopedic treatment, followed by 4% paraformaldehyde fixing about 1 hour, demineralization with 15% EDTA and 0.5% paraformaldehyde for 48 hours, distilled water washing for a night, dimethylbenzene transparant dealing, and paraffin wax embeding. Proliferating Osteoblasts in all these sutures were investigated using immunohiostochemical technique with monoclonal antibodies of proliferating cell nuclear antigen (PCNA). RESULTS: Seven days after orthopedic treatment, no significant difference was observed between the experimental group and the two controlled groups. While after fourteen days, obvious PCNA-positive expression were observed in cells of all these sutures of the experimental group. CONCLUSION: The distribution of proliferating cells and the degree of cell proliferation change after premaxillary orthopedic treatment. And significant cell proliferation is observed in the experimental group, but there are no significant differences between the two controlled groups. PMID- 12539541 TI - [A study on the intrinsic compensative mechanism of craniofacial soft and hard tissue structures of Uygur adults with normal occlusion]. AB - OBJECTIVE: To study the intrinsic compensative mechanism of craniofacial soft and hard tissue structures of Uygur adults with normal occlusion and establish standard cephalometric values to evaluate craniofacial soft and hard tissue structures of normal Uygur adults. METHODS: Lateral cephalometric radiographs of 93 Uygur adults with normal occlusion (43 men and 50 women) were evaluated with the same standard. Their age ranged from 18 to 25 years old, and they had normal and well-balanced faces without receiving any orthodontic treatment. Based on the study of craniofacial soft and hard tissue structures with normal occlusion, the horizontal tendency, the vertical tendency pattern and the medial pattern of craniofacial soft and hard tissue structures were compared, and the intrinsic compensative mechanism in the inner structures of the horizontal and the vertical tendency patterns was investigated. Finally 21 indexes were analysed by mean squar deviation and Newman-Keuls test. RESULTS: The obvious compensative changes in the horizontal and vertical tendency patterns were found in teeth, skeleton and soft-tissues. 1. When the horizontal tendency was compared with the medial pattern, the value of ODI and angles of FH-NA and FH-NB increased (P < 0.01), while the angle of SN-PP decreased (P < 0.01). It showed that the anterior maxilla rotated in the counter-clockwise direction, and the anterior mandible rotated in the clockwise direction. The angle of U1-L1 increased (P < 0.01), which meant that the inclination of anterior teeth decreased. 2. When the vertical tendency was compared with the medial pattern, the value of ODI decreased (P < 0.01), while the height of ANS-Me, U1-PP, L1-MP, and the thickness of LL-SnPg increased (P < 0.01); The angle of L1-MP decreased (P < 0.05), while the length of Sn-Stms increased (P < 0.05). These results showed that there was an obvious positive relationship between hard and soft tissues. The soft tissue thickness could compensate the changes of hard tissues to keep the facial patterns harmony. CONCLUSION: Normal occlusion depends on the homeostasis of teeth, skeleton and soft tissues, but doesn't only depend on the normal craniofacial soft and hard tissue structures respectively. The intrinsic compensative mechanism plays an important role in forming normal occlusion, therefore, doctors should choose the best treatment to obtain optimal individual intrinsic compensative mechanism and achieve an optimal balance between the craniofacial structures and the occlusion. PMID- 12539542 TI - [Dental caries in 5-year-old children in Yunnan in 1995]. AB - OBJECTIVE: To investigate the deciduous tooth caries experience of 5-year-old children in Yunnan province. METHODS: Random samples of 5 year-old children from three cities and three counties of Yunnan were surveyed. RESULTS: The caries incidence of 5-year-old children in Yunnan is 75.75%. The mean dmft is 4.44. The filling rate is only 6.57%. CONCLUSION: The deciduous tooth caries incidence of 5 year-old children is high in Yunnan. There are two factors to be considered: 1. The dental health care education to the parents and teachers should be enforced. 2. In order to get the earlier diagnosis and treatment, regular examination and prevention method should be conducted. PMID- 12539543 TI - Relationship between the increase of secretion of sTNF alpha induced by lipopolysaccharides and the enhanced expression of TACE mRNA in HL-60 cells and adhesive cells from human spleen. AB - In order to find out the potential modulators which influence the secretion of TNF alpha, the relationship between the amount of secreted TNF alpha (sTNF alpha) and the level of TNF alpha converting enzyme (TACE) gene expression was studied before and after the stimulation by lipopolysaccharides (LPS) to HL-60 cells and adhered cells isolated from human spleen using cytological and molecular biology techniques and methods (RT-PCR, Dot-blot hybridization, etc.). The experimental results showed that: (1) LPS could induce the increase of expression of TNF alpha mRNA and TACE mRNA, reaching the peak value at 6 h and 10 h respectively after addition of LPS into cell culture medium; (2) The anti-sense oligodeoxyribonucleotide (A-ODN) complementary to TACE mRNA sequence really inhibited the secretion of TNF alpha as a result of blocked translation of TACE gene; (3) Furthermore, it was also observed that RDQ, a kind of injection derived from Chinese traditional herb, had strongly inhibitory effects on the expression of TACE mRNA and secretion of sTNF alpha stimulated by LPS. The above results suggested that the TACE indeed involved in delivering and processing of pro-TNF alpha during the period of LPS stimulation and the study about the regulator/inhibitors of TACE gene expression would be very important to develop new types of therapy agents against toxic and side effects of sTNF alpha during the period of infection/inflammation to human body. PMID- 12539544 TI - Construction and in vitro expression of Streptococcus mutans surface protein encoding DNA vaccine. AB - DNA vaccine plasmids were constructed that encoded two highly-conservative regions of a surface protein, PAc, from the human major cariogenic bacterium, Streptococcus mutans. Antigen expression was evaluated in vitro by immunohistochemical analysis of human endothelial cells following cationic liposome-mediated transient transfection with recombinant plasmid. The results of this study provided a basis for further testing of these recombinant plasmids in primates and for efficacy testing of dental caries DNA vaccines in human volunteers in future. PMID- 12539545 TI - Study on the preparation of recombinant human HSP70 and its presenting-antigen function. AB - The preparation of recombinant human HSP70 and its presenting-antigen function were investigated. Cultured in glucose-free M9ZB medium and induced with IPTG and lactose at a final concentration of 0.02 mmol/L and 5 mmol/L respectively, the engineered bacteria carrying expression vector of human HSP70 gene expressed rHSP70 at an efficiency fo 60%. After the purification with DEAE ion-exchange chromatography, HSP70 with a purity of higher than 90% was obtained. The purified product could bind tumor-antigen peptide in vitro, and the binding was identified by native PAGE containing 5% glycerol. HSP70-peptide complex could activate lymphocytes to produce specific cytotoxicity to tumor cells, suggesting that the recombinant human HSP70 could be used as an antigen-presenting reagent in tumor therapy. PMID- 12539546 TI - Establishment of in vitro cellular model predicting histocompatibility in allograft. AB - A novel in vitro cellular model producting recipient-donor histocompatibility in allograft was developed to select the donor validity. Fifteen couples of blood samples of donor and recipient in human BMT were examined using the model, and skin allograft in mice was performed to test the model. The results showed that the less the differences of histocompatibility evaluated by the model were, the later GVHR in human BMT occurred and the longer the survival time of skin allografts in mice. It was suggested that the model could be used to predict correctly histocompatibility between donor and recipient. PMID- 12539547 TI - Experimental study of serum substantia nigra neuron autoantibody and its effect in Parkinson disease patients. AB - To investigate the serum substantia nigra neuron autoantibody and its effect in the patients with Parkinson disease (PD), substantia nigra slices and a rat model of injection of serum from PD patients in unilateral side substantia nigra were applied. The results showed that the positive rate of substantia nigra neuron autoantibody in PD patients was significantly higher than in the healthy control group (36.67% vs 6.67%, P < 0.01), but no significant difference was found between PD group and myasthenia gravis (MG) group (26.67%, P > 0.05). The sera from PD patients positive for substantia nigra neuron autoantibody could decrease the number of the dopaminergic neurons more seriously than those from MG and the healthy once respectively (both P < 0.01). The results suggested that the immunological mechanism might partly play a role in the development of PD. PMID- 12539548 TI - BCL-1 rearrangement in acute lymphocytic leukemia and its clinical significance. AB - BCL-1 rearrangement (BCL-1/IgH gene rearrangement) in acute lymphocytic leukemia and its clinical significance was investigated. In 38 patients with acute lymphocytic leukemia (ALL), the genomic DNA of mononuclear cells isolated from peripheral blood and bone marrow was amplified by using hemi-nested polymerase chain reaction (PCR) technique and the expression of cyclin D1 protein of mononuclear cells was detected by using immunohistochemical method. Ten patients with acute granulocytic leukemia, 2 with chronic granulocytic leukemia and 10 with normal bone marrow served as control group. The results showed that BCL-1 rearrangement was detectable in 3 of 38 ALL patients (7.9%) and cyclin D1 protein positive expression was detected in 4 ALL patients (10.5%). Three ALL patients with BCL-1 rearrangement were all B-cell leukemia (B-ALL) and accompanied by cyclin D1 protein expression. No BCL-1/IgH rearrangement or cyclin D1 protein expression was detected in 12 patients with granulocytic leukemia and 10 cases of normal bone marrow. Leukocyte counts in peripheral blood of B-ALL patients with BCL-1 rearrangement and (or) cyclin D1 protein expression were significantly increased and the patients had bad reaction to chemotherapy. It was concluded that: 1) BCL-1/IgH gene rearrangement were detected in acute B lymphocytic leukemia; 2) B-ALL patients with BCL-1 rearrangement and (or) cyclin D1 protein expression had poor prognosis. PMID- 12539549 TI - Investigation on marking method for phenomenon on regrowth drug resistance in relapsed acute myelogenous leukemia. AB - A method for evaluating the regrowth drug resistance in relapsed acute myelogenous leukemia (AML) was developed. Drug sensitivity and proliferation of leukemic cells in vitro were determined using leukemic cell colony forming unit (CUF-L), MTT drug-sensitive test, percentage of S phase cells in cell cycle (S%), fluorescent index (FI) and drug resistant index (DRI) by detecting intracellular daunorubicin, expression of P-170 glycoprotein by APAAP assay, and abundance of Bcl-XL mRNA by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) methods. First, the correlation between scoring criteria and cell drug resistance and cell proliferation was investigated in newly untreated AML patients. Second, 20 patients with relapsed AML were marked. According to each tested result, its point(s) was scored. The results showed that among the 20 cases of relapsed AML, 9 were diagnosed as having regrowth drug resistance. It was concluded that the scoring method for regrowth drug resistance was first developed in AML. There was regrowth drug resistance in relapsed AML; clinically circumventing it would be of extreme significance for establishment of new approaches to the treatment in AML. PMID- 12539550 TI - Effect of ligustrazine on hematopoiesis in bone marrow transplantation mice. AB - The effect of Ligustrazine on the hematopoiesis after bone marrow transplantation (BMT) in allogenic BMT mice was investigated. After the typical mice model of allogenic BMT had been established, the mice were randomly divided into three groups: BMT group, Ligustrazine group and normal group. The BMT group was given normal saline (0.2 ml, twice a day) through gastric tube, while the Ligustrazine group was given Ligustrazine through gastric tube (0.2 ml, twice a day). At the 1st, 7th and 14th day after BMT, we observed the peripheral blood cells and bone marrow nuclear cells (BMNC), as well as the expression level of Heparan Sulfate (HS) and stromal cell derived factor-1 (SDF-1) on bone marrow sections by using immunohistochemistry (SABC-AP), the expression of CXCR4 on the BMNC. The results showed that on the 7th and 14th day, the peripheral blood white cells, platelets, BMNC and the expression levels of CXCR4, HS and SDF-1 were significantly higher in Ligustrazine group than in the BMT group (P < 0.05). It was concluded that Ligustrazine could promote hematopoiesis at the early stage of hematopoietic reconstitution after BMT. PMID- 12539551 TI - An experimental study on the regulation of expression of Th2 cytokines from T lymphocytes by protein kinase C in asthma. AB - To explore the regulatory role of protein kinase C (PKC) in the expression of Th2 cytokines, interleukin-4 (IL-4) and interleukin-5 (IL-5) by T lymphocytes in asthma. T lymphocytes were isolated and purified from blood and bronchial alveolus lavage fluid (BALF) of each guinea pig of normal control group and asthmatic group and from peripheral blood of the asthmatic patients and normal controls, and were stimulated with PKC accelerant phorbol 12-myristate 13-acetate (PMA) and inhibitor Ro31-8220. The expression of IL-4 and IL-5 mRNA and protein was detected by using in situ hybridization staining and ELISA respectively. The expression of IL-4 and IL-5 mRNA and protein of asthmatic T lymphocytes stimulated with PMA was significantly higher than that of asthmatic T lymphocytes stimulated without PMA respectively (P < 0.01) and that of normal T lymphocytes stimulated with PMA respectively (P < 0.01). The expression of IL-4 and IL-5 mRNA and protein of asthmatic T lymphocytes stimulated with PMA and Ro31-8220 was significantly lower than that of asthmatic T lymphocytes stimulated only with PMA respectively (P < 0.01). It was concluded that PKC might participate in regulating the expression of IL-4 and IL-5 in asthmatic T lymphocytes, and the activation of PKC in T lymphocytes might play an important role in the pathogenesis of asthma. PMID- 12539552 TI - mRNA expression of basic fibroblast growth factor from a single intratracheal instillation of papain-induced emphysema in rats. AB - The relations between mRNA expression of basic fibroblast growth factor (bFGF) and the changes in collagen I and collagen III in pulmonary tissues from a single intratracheal instillation of papain-induced emphysema in rats were investigated. Wistar rats (n = 42) were randomly divided into normal group and emphysema model 1, 3, 5, 7, 15, 30-day groups (n = 6 in each group). The rat model of emphysema was induced by a single intratracheal instillation of papain. The results of immunohistochemistry SABC and in situ hybridization with bFGF probe were quantitatively analyzed to examine the changes of collagen I and collagen III and bFGF mRNA expression in lung tissues and the percent of positive expression of bFGFmRNA in alveolar macrophages. The results were as follows: (1) In the emphysema model groups the optical densities of collagen I and collagen III began to increase after 3 days, reached the highest at the 7th day, and began to reduce at the 15th day; (2) No expression of bFGFmRNA in pulmonary tissues was detectable in the normal group. The positive expression of bFGFmRNA was detectable in lung tissues one day after the intratracheal instillation of papain. The average optical densities reached the peak (41.895 +/- 7.017) at the 7th day, significantly higher than in the normal group (0.581 +/- 0.139, P < 0.01). The positive expression of bFGFmRNA in lung tissues began to reduce at the 15th day; (3) Positive expression of bFGFmRNA in alveolar macrophages of instillation papain rats was detectable 3 days after the intratracheal instillation of papain, and reached the highest at the 7th day with the percent of positive expression of bFGF mRNA in alveolar macrophages being 70.13 +/- 11.21, higher than in the normal group (5.12 +/- 0.18, P < 0.01); (4) The expression of bFGF mRNA in the lung tissues and macrophages was positively related with the changes in collagen I and collagen III (P < 0.01 or P < 0.05) respectively. It was suggested that the up-regulation of bFGF mRNA expression during the development of emphysema can lead pulmonary interstitial fibrosis, which may take part in the injury and repair and the lung tissue reconstruction. PMID- 12539553 TI - The study on the relationship between serum vascular endothelial growth factor and proteinuria in adriamycin-induced nephrotic rats. AB - To study the relationship between serum vascular endothelial growth factor (VEGF) and proteinuria in adriamycin-induced nephrotic rats, a rat model of adriamycin induced nephrotitis was developed by injection of adriamycin into a tail vein in a rat. At different time points, 24-h urinary protein excretion was measured by using Coomassie brilliant blue method and the serum VEGF levels detected by using ELISA assay. The interventional effect of VEGF on this model was observed. The results showed that: (1) The adriamycin-induced nephrotic syndrome rat model was developed successfully; (2) Serum VEGF levels and proteinuria were significantly increased at 7th day after intravenous injection of adriamycin. There was a positive correlation between serum VEGF levels and 24-h urinary protein excretion (r = 0.67, P < 0.05). (3) The 24-h urinary protein excretion was significantly increased in the rats receiving administration of VEGF (P < 0.05). It was concluded that VEGF might play an important role in the pathogenesis of proteinuria in adriamycin-induced nephrotic rats. PMID- 12539554 TI - The experimental study on the effect calcitonin gene-related peptide on bone resorption mediated by interleukin-1. AB - To investigate the effect of calcitonin gene-related peptide (CGRP) on bone resorption mediated by interleukin-1 beta(IL-1 beta) in vitro, the osteoclasts isolated from the long bones of newborn SD rats were co-cultured with osteoblasts on ivory slices placed in 24-well plates. 24 h later, conditioned media containing CGRP and/or IL-1 beta were added to the wells respectively, and continued culturing for 48 h. After the cells were stripped off by ultrasonication, the ivory slices were stained in toludine blue. The number and the total area of resorption lacunae on each slice were measured by computer imaging analysis system. Our results showed that IL-1 beta significantly stimulated bone resorption, but CGRP inhibited the effect mediated by IL-1 beta in a dose-dependent manner. It is suggested that CGRP may inhibit osteoclastic bone resorption through two ways: One is that CGRP functions directly on osteoclasts to block their activation; the other is that CGRP regulates the release of cytokines by osteoblasts and indirectly affects the function of osteoclasts. PMID- 12539555 TI - Clinical significance and assessment of cytokines in various stages of ulcerative colitis. AB - In order to study the clinical significance and change of interleukin (IL)-1 beta and IL-10 concentration in intestinal mucosal tissues in various stage of ulcerative colitis (UC), IL-1 beta and IL10 levels were measured by enzyme linked immunosorbent assays (ELISA). Our results showed that IL-beta level caused by spontaneous secretion in the intestinal mucous tissues in active stage of ulcerative colitis was significantly higher than that in normal controls and in remission stage of ulcerative colitis (P < 0.01, P < 0.001). IL-10 level in various stage of UC was relatively lower in controls, but there was no significantly difference between the two groups. Our study suggested that higher IL-1 beta level in active might play an important role in pathogenesis of UC, and IL-10, as an anti-inflammatory cytokine, was low in active UC, suggesting that it may be a important factor contributing to the development of higher IL-1 beta level. PMID- 12539556 TI - Effect of xiaokuiling prescription on the expression of HSP72, HSP B in gastric mucosa of patients with Helicobacter pylori-associated duodenal ulcer. AB - In order to investigate the mechanism of Xiaokuiling prescription (XKL) in the treatment of Helicobacter pylori (HP)-associated duodenal ulcer (DU) and the pathophysiologic role of heat shock proteins (HSPs) in the healing of ulcer, the expression of HSP72 and HSP B in gastric mucosa was detected by using SABC immunohistochemistry method and processed by micro-image analysis system. The method of Western blotting was used to measure the contents of HSP72 and HSP B in the tissue emulsion of gastric mucosa. The results were as follows: (1) HSP72 expression of the gastric mucosa in the treated group was obviously increased as compared with that in the control group (P < 0.05); (2) HSP B expression of the gastric mucosa in the treated group was significantly decreased as compared with that in the control group (P < 0.01). It was suggested that the increased expression of HSP72 and the elimination of HP might be related to the mechanism of action of XKL. HSPs might play an pathological and physiological role in the process of healing of gastric ulcer. PMID- 12539557 TI - Molecular tissue engineering: applications for modulation of mesenchymal stem cells proliferation by transforming growth factor beta 1 gene transfer. AB - The effect of transforming growth factor beta 1 (TGF-beta 1) gene transfection on the proliferation of bone marrow-derived mesenchymal stem cells (MSCs) and the mechanism was investigated to provide basis for accelerating articular cartilage repairing using molecular tissue engineering technology. TGF-beta 1 gene at different doses was transduced into the rat bone marrow-derived MSCs to examine the effects of TGF-beta 1 gene transfection on MSCs DNA synthesis, cell cycle kinetics and the expression of proliferating cell nuclear antigen (PCNA). The results showed that 3 microliters lipofectamine-mediated 1 microgram TGF-beta 1 gene transfection could effectively promote the proliferation of MSCs best; Under this condition (DNA/Lipofectamine = 1 microgram/3 microliters), flow cytometry and immunohistochemical analyses revealed a significant increase in the 3H incorporation, DNA content in S phase and the expression of PCNA. Transfection of gene encoding TGF-beta 1 could induce the cells at G0/G1 phase to S1 phase, modulate the replication of DNA through the enhancement of the PCNA expression, increase the content of DNA at S1 phase and promote the proliferation of MSCs. This new molecular tissue engineering approach could be of potential benefit to enhance the repair of damaged articular cartilage, especially those caused by degenerative joint diseases. PMID- 12539558 TI - Effect of transforming growth factor-beta 2 on phagocytosis in cultured bovine trabecular meshwork cells. AB - The effect of transforming growth factor-beta 2 (TGF-beta 2) on phagocytosis in bovine trabecular meshwork cells in vitro was investigated. After the cultured bovine trabecular meshwork cells were treated with 0 ng/ml, 0.32 ng/ml, 1 ng/ml, 3.2 ng/ml TGF-beta 2 for 24 h, latex beads were added into the incubation medium, and the numbers of the latex beads in 20 adjacent cells were counted under a microscope 24 h later, after treatment with Wright's stain. Our results showed that the average numbers of the latex beads in the trabecular meshwork cells treated with TGF-beta 2 of different concentrations were 53.1 +/- 1.7 beads/cell, 56.4 +/- 2.9 beads/cell and 77.9 +/- 6.5 beads/cell respectively, in comparison with 45.5 +/- 3.3 beads/cell of the control group. TGF-beta 2 significantly increased the number of the latex beads phagocytosed by cultured bovine trabecular meshwork cells in a dose-dependent manner. TGF-beta 2 could promote the phagocytosis of bovine trabecular meshwork cells in vitro. It may be involved in the cellularity decrease of the trabecular meshwork in the patients of primary open angle glaucoma through promoting the phagocytosis of trabecular meshwork cells. PMID- 12539559 TI - Study on the changes in microvessel density in hepatocellular carcinoma following transcatheter arterial chemoembolization. AB - To study the changes in intratumoral microvessel density (MVD) in hepatocellular carcinoma (HCC) following transcatheter arterial chemoembolization (TACE), MVD in 42 HCC specimens histologically verified was studied by using immunohistochemical method. Of all the specimens, 20 were obtained from the patients treated with surgical resection alone, 22 from those with second stage surgical resection after TACE. The results showed that the MVD in HCC tissues was 53.4 +/- 21.9 in the TACE group and 27.6 +/- 9.2 in the single operating group, respectively, with the difference being significant between them (P < 0.001). It was suggested that TACE might contribute to angiogenesis of HCC, possibly due to anoxic stress and ischemia-reperfusion injury. PMID- 12539560 TI - The influence of enteral nutrition on gut barrier in the post-operative patients with damaged hepatic function. AB - The safety, rationality and the practicality of enteral nutrition (EN) support in the postoperative patients with damaged hepatic function were investigated and the protective effect of EN on the gut barrier and the clinical implication studied. Seventy-six adult patients whose hepatic function were in Child B or C grade were randomly assigned in EN group (30 cases), total parenteral nutrition (TPN) group (26 cases) and control group (CON, 20 cases). The patients received different nutritional support. The signs of nutritional condition and hepatic function were messured at 1 day before, 5 days and 10 days after the surgical operation respectively. The changes in the urine lactulose (L) and mannitol (M) contents and L/M ratio were observed by using pulsed electrochemical detection (HPLC-PED) to acquire the different effects among the different nutritional support performance. The results showed that the patients in the EN group and TPN group had no worse hepatic function damage after operation. The patients in the EN group reached the positive nitrogen balance earlier, had a less weight loss than in the TPN group with the difference being significant (P < 0.05). There was no obvious change in L/M ratio in the postoperative patients in the EN group (P > 0.05), but there was significant difference in L/M between TPN group and CON group (P < 0.05). It was concluded that EN was a rational, safe, effective and practical nutrition support method in the patients with damaged hepatic function patients after surgical operation and EN can effectively protect the structure and function of gut barrier from sever infection. PMID- 12539561 TI - Effect of yiqitongyanghuatan recipe on experimental atherosclerosis in rabbits. AB - In order to investigate the roles of Yiqitongyanghuatan (YQTYHT) recipe in reducing the levels of serum cholesterol and plasma lipid peroxidation (LPO), platelet aggregation function (PAgF) and platelet adhesion function (PAdF), the area of atherosclerotic plague coverage in aorta and the thickness of plague, 32 male Japanese white rabbits were divided into 4 groups. The results showed that the YQTYHT recipe could significantly lower the levels of serum cholesterol and tryglyceride, plasma LPO, and PAgF and PadF. The area of atherosclerotic plague coverage in aorta and the thickness of plague in the YQTYHT-fed rabbits were decreased as compared with that in the high-cholesterol-fed rabbits. The above roles might contribute to the main mechanism of YQTYHT against atherosclerosis. PMID- 12539562 TI - Gene analysis of free fetal DNA in maternal plasma. AB - To investigate the feasibility of using free fetal DNA from maternal plasma as the source of fetal material in non-invasive prenatal diagnosis, SRY gene of free DNA in maternal blood of 65 samples were analyzed by using primer extension preamplication (PEP) and probe microplate hybridization techniques. The results showed that the detection rate of SRY gene in maternal blood from women carrying male fetuses detected by probe microplate hybridization alone and probe microplate hybridization with PEP were 76.09% (35/46) and 95.65% (44/46) respectively, and there was a significant difference between them. The non detection rate of SRY gene in blood samples from women carrying female fetus was 100% (19/19). It is indicated that probe microplate hybridization was an effective method in detecting trace fetal DNA from maternal plasma and the sensitivity could be substantially improved by combined use of the two techniques. Analysis of fetal DNA in maternal plasma can serve as an alternative for non-invasive prenatal diagnosis. PMID- 12539563 TI - Changes in serum leptin levels during r-hGH treatment in growth hormone-deficient children. AB - To observe the effect of growth hormone on serum leptin levels, serum leptin concentrations were measured by enzyme immunoassay in 12 prebuttal children with growth hormone deficiency 1, 3 and 6 months before and after the treatment with recombinant human growth hormone (r-hGH). For comparison, 34 normal prepubertal children were also investigated. Relationship between leptin levels and body mass index (BMI) was observed at the same time. Our results showed that serum leptin level in normal prepubertal children was 1.22 +/- 0.34 ng/ml; the pretreatment serum leptin levels in GHD children was 3.08 +/- 2.41 ng/ml, which was significantly different from those 1, 3 and 6 months after GH treatment (i.e. 1.64 +/- 1.37 ng/ml, 1.57 +/- 1.40 ng/ml and 1.35 +/- 0.89 ng/ml respectively) (all P < 0.001). Our results suggested that r-hGH has a suppressive effect on leptin expression. PMID- 12539564 TI - Clinical characteristics of transmitted transfusion virus infection in children. AB - Clinical characteristics of transmitted transfusion virus (TTV) infection and its pathogenicity in children were evaluated. Serum TTV DNA from 118 children (mean age: 7.8 +/- 2.8 years) was detected by nested PCR. The product of PCR was cloned and sequenced. The positive rate for serum TTV-DNA in 20 healthy children, 9 cases of acute hepatitis, 51 cases of chronic hepatitis, 24 cases of nephritis or nephrotic syndrome and 14 cases of hypoplastic anemia or acute leukemia was 20%, 11%, 29%, 42% and 21% respectively, but there was no significant difference in TTV-DNA frequency among them (P > 0.05). Of the 16 patients receiving immunosuppressive agent for a long time, 7 (44%) were positive for TTV-DNA, and of the 17 cases not receiving immunosuppressive agent, 5 (29%) were positive with the difference being not significant (P > 0.05). Essential characteristics were pathogen-carrier or asymptomatic infection in children with TTV infection. Long term employment of immunosuppressive agent did not increase the incidence in TTV infection. There was still high prevalence in TTV infection in healthy children not receiving blood product, suggesting the possibility of non hematogenous transmitted transfusion in TTV transmission. PMID- 12539565 TI - In vitro validation of tissue Doppler left ventricular regional wall velocities by using a novel balloon phantom. AB - To investigate the validity and accuracy of tissue Doppler imaging (TDI) using a novel balloon phantom, validation of TDI myocardial velocity measurements has been carried out indirectly from conventional M-mode images. However it is not a true and independent gold standard. We described a new TDI validation method by using a specially developed left ventricular balloon model mounted in a water bath and constructed using two pear-shaped balloons. It was connected to a pulsatile flow pump at 8 stroke volumes (50-85 ml/beat). The displacement and velocity of the balloon walls were recorded simultaneously by video imaging and TDI on a GE-Vingmed System Five with a 5 MHz phased array probe at the highest frame rates available. Conventional M-mode and 2-D imaging verified that our balloon model mimicked the shape and wall motion of left ventricle. There was a good correlation and agreement between the maximum video excursion of the anterior and posterior walls of the phantom and the results of the temporal integration of digital distance data by TDI (Anterior wall: r = 0.97, SEE = 0.24 mm, mean +/- s = 0.04 +/- 0.24 mm; Posterior wall: r = 0.95, SEE = 0.22 mm, mean +/- s = 0.03 +/- 0.24 mm). Analysis of the velocity profile by the TDI method showed that the velocity at each measured point was correlated well with the velocity obtained from the video images (Anterior wall: r = 0.97, SEE = 0.30 mm, mean +/- s = -0.04 +/- 0.28 mm; Posterior wall: r = 0.97, SEE = 0.30 mm, mean +/- s = 0.04 +/- 0.28 mm). Our balloon model provided a new independent method for the validation of TDI data. This study demonstrated that the present TDI system is reliable for measuring wall motion distance and velocity. PMID- 12539566 TI - A preliminary clinical study of three-dimensional ultrasonography in prenatal diagnosis. AB - To evaluate the clinical value of three-dimensional ultrasonography (3DUS) in prenatal diagnosis, 134 pregnant women with high-risk factors in second and third trimester were examined by 3DUS. The results showed that 3DUS could provide more diagnostic information, exclude the abnormalities and enhance the confidence level of physician in 102 normal pregnant women. 3DUS was helpful in the diagnosis in 17 (60.7%) of 28 cases of fetal anomalies. However, 3DUS was not useful in evaluating intrauterine growth retardation in 4 cases. It is concluded that 3DUS is helpful in prenatal diagnosis. PMID- 12539567 TI - Pulmonary sequestration: three dimensional dynamic contrast-enhanced MR angiography and MRI. AB - In order to evaluate the diagnostic value of three-dimensional contrast-enhanced MR angiography and MRI for pulmonary sequestration, 5 patients with pulmonary sequestration underwent 3D fast imaging by steady state precession (FISP) with a contrast medium and breath holding following chest radiography, CT and MR scans. The reconstructed MR angiography was performed using maximum intensity projection (MIP) and multiplanar reconstruction (MPR) techniques. It was found that the chest radiography showed pulmonary sequestration as a persistent area of opacity in the posterior basal segment of the left lower lobe, which was close to mediastinum in 2 cases and close to diaphragma in 3 cases. CT revealed a soft issue mass beyond descending aorta and lobar emphysema around the pulmonary sequestration. And the supplying vessel was documented in 2 cases on enhanced CT. MRI demonstrated a hyperintensity mass with respect to normal lung parenchyma on T1WI and T2WI, and the origin of the supplying vessel in 3 cases. The reconstructed CE MRA using MIP or MRP techniques clearly showed the supplying vessel and its course, branches as well as draining vessels. It was concluded that 3D CE MRA of demonstrating the supplying and draining vessels to pulmonary sequestration, together with plain MRI, can provide a diagnosis and aid in surgical planning without the need for DSA. PMID- 12539568 TI - Potent antioxidative potential of propofol during cardiopulmonary bypass in the adult. AB - The potent antioxidative potential of propofol during cardiopulmonary bypass (CPB) in adults was investigated. The selected 30 patients receiving open heart surgery under CPB were randomly divided into group A and group B. The patients in the group A and group B were given propofol (0.1 mg.kg-1.min-1) and fentanyl (5 micrograms.kg-1.min-1) respectively to maintain anesthesia after aorta was cross clamped. Blood samples were drawn pre-anesthesia, pre-CPB, at 30 min of CPB, at the end of CPB, at 1 h after CPB, at the end of operation, at 12 and 24 h postoperatively. RBC suspension was prepared and erythrocyte glucose-6-phosphate dehydrogenase (G-6-PD) and phosphofructokinase (PFK) activities, total erythrocyte reduced glutathione (GSH) and oxidized GSH (GSSG) were assayed and GSH/GSSG ratio was calculated. In the group A, G-6-PD and PFK activities and GSH/GSSG ratio were almost uneventfully during CPB and postoperatively. In the group B, G-6-PD activity was increased and PFK activity and GSH/GSSG ratio decreased significantly from 30 min of CPB until 12 h postoperatively. It was demonstrated that propofol could obviously attenuate free radical activity during CPB, while fentanyl has no effect on free radical reduction. Propofol could be beneficial as an anesthetic in patients presenting pathologies associated with free radical reactions during CPB. PMID- 12539569 TI - The binding ability analysis of the normal VLDL receptor and its mutant. AB - The ligand-binding domain of VLDL receptor contains eight imperfectly similar repeats. To discuss the contribution of each repeat to ligand binding, the RT-PCR technique was used to clone the VLDLR-cDNA from the heart muscle of Chinese people. Two recombinants were further constructed, which contained the full length cDNA of VLDLR and the mutant lacking repeats 1-5. CHO cell line was transfected with two recombinants. The expression of VLDLR gene could be detected by RT-PCR from the CHO cells transfected with pCD-VR. The results of binding experiments showed that the ability of the CHO cells transfected with the full length cDNA of VLDL-R binding DiI-labeled beta-VLDL was higher than that of the CHO cells transfected with the mutant. Our findings indicated that human VLDL-R gene could be expressed effectively on CHO cells, and the receptor was almost inactivated when repeats 1-5 were deleted. PMID- 12539570 TI - Comparative study on the immunogenicity between Hsp70 DNA vaccine and Hsp65 DNA vaccine in human Mycobacterium tuberculosis. AB - The BALB/c mice were immunized with Hsp70 DNA and Hsp65 DNA vaccines in human Mycobacterium tuberculosis. Eight weeks after immunization, the eyeballs were removed, blood and spleen taken, and intraperitoneal macrophages were harvested. The lymphocytic stimulating index (SI) was used to measure the cellular proliferating ability and NO release to measure the phagocytic activity of the macrophages. With ELISA kit, the levels of interleukin-2 (IL-2) and interferon gamma (IFN-gamma) in serum and the splenic lymphocytic cultured supernatant were detected. The results showed that after the mice were immunized with 100 micrograms/mouse of Hsp70 DNA vaccine intramuscularly, the splenic lymphocytic proliferating ability in the mice was significantly increased as compared with that in the control group, vector group and Hsp65 DNA vaccine group (P < 0.01); The contents of NO in the intraperitoneal macrophages of the mice were significantly lower than in the control group and Hsp65 DNA vaccine group (P < 0.01); The levels of serum IL-2 in the mice were significantly higher than in the control group, but there was no statistical difference between Hsp65 DNA group and vector group (P > 0.05); The contents of serum IFN-gamma in the mice were significantly higher than in the control group, but significantly lower than in the Hsp65 DNA vaccine group (P < 0.05). It was indicated that immunization with Hsp70 DNA vaccine could obviously enhance the immune response, but its intensity seemed inferior to Hsp65 DNA vaccine. The anti-infection mechanisms and clinical use in the future of the vaccines of Hsp70 DNA and Hsp65 DNA are worth further studying. PMID- 12539572 TI - Follicle-stimulating hormone (FSH) induced internalization of porcine FSH receptor in cultured porcine granulosa cells and Chinese hamster ovary cells transfected with recombinant porcine FSH receptor cDNA. AB - In order to study the fate of human follicle-stimulating hormone (FSH) when hormone binds to its receptor, a quick biochemical method that can differentiate between the surface-bound and internalized hormone was used to determine the internalization induced by FSH in cultured both porcine granulosa cells and Chinese hamster ovary (CHO) cells expressing recombinant porcine FSH receptor. The results showed that FSH was slowly internalized, and the internalized radioactivity (acid resistant) reached a peak 10-12 h after addition of 125I hFSH. It was suggested that FSHR do not get internalized rapidly under physiological circumstances precisely because the appropriate sequences are absent. PMID- 12539571 TI - Effects of DDPH on HECCM-induced proliferation and immunophenotypes of the pulmonary vascular pericytes. AB - In order to study the effects of 1-(2, 6-dimethylphenoxy)-2-(3, 4 dimethoxyphenylethylamino) propane hydrochloride (DDPH) on proliferation and immunophenotypes of newborn rat pulmonary vascular pericytes induced by hypoxic endothelial cell conditioned medium (HECCM) from porcine pulmonary arteries, the cultured pericytes were divided into 4 groups according to the endothelial cell conditioned medium (ECCM) used: normoxic ECCM (NECCM) group, NECCM + DDPH group, HECCM group and HECCM + DDPH group. Cell culture, immunocytochemical staining, image analysis and flow cytometric method were used to investigate the effects of HECCM and DDPH on the expression of alpha-smooth muscle actin (alpha-SM-Actin) antigen, CD34 antigen, S-100 antigen and proliferating cell nuclear antigen (PCNA) and cell cycle in pericytes. The results showed that the alpha-SM-Actin antigen in the pericytes in HECCM group was stronger positively expressed than in the other three groups, but CD34 antigen and S-100 antigen were negatively expressed. The expression of alpha-SM-Actin antigen, CD34 antigen and S-100 antigen was positive in the groups of NECCM, NECCM + DDPH and HECCM + DDPH; The expression of alpha-SM-Actin and PCNA in HECCM group was 1.32 times (P < 0.01) and 1.24 times (P < 0.05) that in NECCM group, 1.30 times (P < 0.01) and 1.21 times (P < 0.05) that in HECCM + DDPH group, respectively. The percentage of the cells in the GO-G1 phase in the HECCM group was lower by 11.7% and 9.1%, in S phase higher by 5.6% and 4.2%, in G2-M phase higher by 6.1% and 4.9% than in the groups of NECCM, HECCM + DDPH, respectively. The inhibitory rate of DDPH on the increased alpha-SM-Actin and PCNA syntheses in pericytes induced by HECCM were 23.4% and 17.1% respectively. The inhibitory rate on the increased pericytes from GO-G1 phase to S phase was 8.3%. These results suggest that DDPH can directly inhibit pericytes from proliferation and immunophenotypical transformation of smooth muscle-like cells induced by HECCM. PMID- 12539573 TI - Cloning and identification of a novel variant of human vascular endothelial growth factor. AB - A novel variant of human vascular endothelial growth factor (h'VEGF165) cDNA was amplified by nested PCR method from the HL601 cells and was cloned into a eukaryotic expressing vector pcDNA3 to construct a recombinant plasmid pCD h'VEGF165. The amplified h'VEGF165 cDNA fragment was identified by enzyme digestion and DNA sequencing methods. Also, wild-type hVEGF165 cDNA was obtained, identified and cloned into a eukaryotic expressing vector pcDNA3 by using the same methods. The results of DNA sequencing showed that h'VEGF165 cDNA cloned from HL601 was 600 bp in size with 8% of the base sequence in h'VEGF165 cDNA being changed as compared with the base sequence in the wild-type hVEGF165 cDNA. The results of sequencing of hVEGF165 which was cloned from HL60 by us were consistent with the reports completely. PMID- 12539574 TI - The mechanical study of vascular endothelial growth factor on the prevention of restenosis after angioplasty. AB - The mechanism of vascular endothelial growth factor (VEGF) on the prevention of restenosis after angioplasty was investigated. The cultured vascular endothelial cells (VEC) were incubated with the conditioned medium (CM) from vascular smooth muscle cells (VSMC) infected with recombinant adenoviruses containing the hVEGF165 gene. To observe the effects of VEGF on proliferation and NO, ET, 6-keto PGF1 alpha secretion of VEC, WST-1 method, Griess method and radioimmunoassay were used respectively. The PDGF-B mRNA transcription in VECs was detected by RT PCR. It was showed that NO, 6-keto-PGF1 alpha and OD value were markedly increased in a dose-dependent manner in the VEGF-treated groups as compared with those in the control group, while ET and PDGF-B mRNA were significantly decreased in the VEGF-treated groups (P < 0.05 or P < 0.01). Adenovirus vector mediated hVEGF165 gene could promote the proliferation of VECs and improve NO, PGI2 secretion, inhibit ET secretion and PDGF-B mRNA transcription in the VECs. The above results offered further theoretical evidence for VEGF on the prevention of restenosis after angioplasty. PMID- 12539575 TI - Experimental study on AT1-receptor-peptide-induced myocardial immune damage in rat. AB - In order to investigate the immunological damage in rat immunized with AT1 receptor peptide, 18 male Wistar rats were divided into two groups: immunized group (n = 12), each rat was immunized with 150 micrograms AT1-receptor peptide coupled to bovine serum albumin, together with Freund's adjuvant. Control group (n = 6), sham-immunized, "immunized liquid" was same as immunized-group except AT1-receptor peptide. Systolic blood pressure (SBP) was measured by using the tail-cuff technique, antibody against AT1-receptor peptide detected by using ELISA method, and left ventricular myocardium and renal cortex sections were observed under light and electron microscopy. There was no significant difference in SBP and light microscopic observation of the tissue sections between the immunized-group and control group. The O.D. value of anti-AT1-receptor peptide antiserum was significantly higher in the immunized-group than in the rats before immunization and control group (P < 0.01). Positive rate in the immunized-group was 100%, while 0% in the control group. Ultramicroscopic morphology showed potential myocardial injury, including: increase in number of mitochondria, swelling of many mitochondria with reduction in number or absence of their cristae and cristolysis, disorder of the cardiac myofibrils, and myofibrillar disruption and myocytolysis. And lysosomes were increased in renal tubular epithelia. The AT1-receptor peptide could induce to generate the antibody against AT1-receptor peptide and lead to myocardial and renal damage in rats. PMID- 12539577 TI - The effect of Erigeron breviscapus on proliferation of pulmonary artery smooth muscle cells in hypoxic porcines. AB - In order to study the effect of Erigeron Breviscapus (EB) on proliferation of pulmonary artery smooth muscle cells (PASMC) in hypoxic porcines, immunohistochemical and MTT methods were employed to measure the proliferation of PASMC. It was found that the proliferation of PASMC in porcines was obvious, and the expression of proliferating cell nuclear antigen (PCNA) was significantly high within 48 h after exposure to hypoxia. The EB could inhibit the proliferation and the expression of PCNA in PASMC under hypoxia, but it had no effect on the proliferation and expression of PCNA in PASMC under normal condition. The EB could inhibit the proliferation and the expression of PCNA in PASMC induced by phorbol 12-myristate 13-acetate (PMA), an agonist of PKC in normal and hypoxic conditions. It was concluded that the hypoxia could enhance the proliferation and expression of PCNA in PASMC. The EB can inhibit the proliferation and expression of PCNA in PASMC under hypoxia through PKC-signal way. The EB may be used in treating the pulmonary hypertension by inhibiting the proliferation of PASMC and the pulmonary vascular remodeling. PMID- 12539576 TI - Study on apoptosis and expression of P53, bcl-2, Bax in cardiac myocytys of congestive heart failure induced by ventricular pacing. AB - The apoptosis and the expression of p53, bcl-2 and Bax in myocytes of chronic rapid ventricular pacing-induced congestive heart failure (CHF) in rabbits were investigated. The CHF rabbit model (P, n = 7) was established by chronic rapid ventricular pacing for 3 weeks. By using TUNEL technique the apoptosis in the myocytes in the rabbit model was studied and the expression of p53, bcl-2 and Bax in myocytes was detected by using immunohistochemical method. Sham-operated (C, n = 9) group served as control group. The results showed that there were about 4033 +/- 884.56 apoptotic cells/10(6) myocytes in P group, but no apoptotic cells were found in C group. Myocytes positive for p53 immunoreactivity (18.86 +/- 8.48 vs 5.06 +/- 0.87, P < 0.01) and positive for Bax immunoreactivity (7.15 +/- 1.91 vs 0.43 +/- 0.09, P < 0.01) were increased in P group as compared with those in C group, while the myocytes positive for bcl-2 immunoreactivity (7.08 +/- 1.05 vs 14.97 +/- 4.47, P < 0.01) and the ratio of bcl-2/Bax were decreased in P group as compared with those in C group. Apoptosis was involved in the development of CHF induced by continuously rapid ventricular pacing in rabbit. The expression of p53 and Bax was increased, while the expression of bcl-2 was inhibited. These might play an important role in the acceleration of the apoptosis. PMID- 12539578 TI - Application of CD45/SSC gating multiparameter flow cytometry in the classification of acute leukemia--an analysis of 139 cases. AB - In order to study the significance of flow cytometry immunophenotyping in the diagnosis of acute leukemia, CD45/SSC gating multiparameter flow cytometry (FCM) was utilized to analyze the immunophenotypes of 139 cases of acute leukemia. 139 cases of acute leukemia were enrolled in our hospital from April 1998 to April 2000. Morphological analysis and FCM immunophenotypic tests were conducted on all cases. Our results showed that CD45/SSC gating multiparameter flow cytometry immunophenotyping could reflect the origin of leukemic cells specifically. It is one of the important methods for the diagnosis of ALL, AML, and HAL. CD45/SSC gating multiparameter FCM analysis is a good technique for immunophenotyping. FCM immunophenotypic analysis can help improve the diagnosis and classification of acute leukemia, and extend the use of FCM in clinical practice. PMID- 12539579 TI - The effect of the Fas/FasL pathway during chemotherapeutic drug-induced apoptosis of leukaemeic cells. AB - The mechanism of chemotherapeutic drug-induced apoptosis in leukaemic cells was studied to further investigate whether Fas/FasL system was involved in apoptosis induced by chemotherapeutic drugs and assess their effects when used in combination with soluble FasL (sFasL). The expression of Fas on human leukaemic cell lines K562, HL-60 and U937 treated with daunorubicin (DNR) or cytosine arabinoside (Ara-C) was detected by using flow cytometry. The activities of sFasL, DNR and Ara-C inducing apoptosis of leukaemic cells, in the absence or presence of neutralizing anti-Fas IgG antibody, were detected by using flow cytometry and TUNEL. The results showed that flow cytometric profiles of K562, HL 60 and U937 cells treated with DNR or Ara-C failed to show any significant increase in Fas expression over 18 h (P > 0.05). Anti-Fas monoclonal antibody (IgG) could not block the apoptosis in leukaemic cells induced by DNR or Ara-C, but could block the apoptosis induced by sFasL. A role of sFasL in a cytotoxic synergistic effect when used in combination with chemotherapeutic drugs was revealed. It was concluded that chemotherapeutic drug-induced apoptosis in human leukaemic cells (UG37, HL-60) is independent of the Fas/FasL system, but combination of sFasL and drug treatment produces a synergistic cytotoxic effect on human leukaemic cells. PMID- 12539580 TI - Bioactivities of culture supernatants from retroviral packaging cells carrying the mouse Fas ligand gene. AB - The bioactivities of culture supernatants from retroviral packaging cells carrying the mouse Fas ligand (mFasL) gene was investigated. FasLcDNA was cloned into PLXIN with an internal ribosome entry site to link two cistrons through gene recombination technology, PLXIN and the recombinant vector PLFIN were separately transfected into PA317 retrovirus packing cell line by lipofectamine 2000, and the resistant clones were selected with G418 selective medium. The integration of genome DNA was assayed by genomic DNA PCR. NIH3T3 cells were transduced by the culture supernatants from PA317 carrying the mFasLcDNA gene, and were selected with G418 selective medium, so as to select the PLFIN-PA317 clone capable of producing higher titer of supernatants. The levels of mFasL protein on NIH3T3 cells membrane were assayed by flow cytometry (FCM). The biological activity of mFasL on NIH3T3 cells membrane was investigated by the inducing apoptosis of Fas+ Yac-1 cells co-cultured with NIH3T3 cells expressing Fas ligand. To explore the direct mFasL cytotoxicity of culture supernatants from retroviral packaging cells carrying the mFasL gene, the culture supernatants from PLFIN-PA317 and PLXIN PA317 were separately co-cultured with Yac-1 cells in parallel. The recombinant PLFIN was successfully constructed. The highest titer of supernatants from twelve resistant clones was 8.5 x 10(5) colony-forming-unit (CFU)/ml. The NIH3T3 cells transfected by above supernatants had a higher level of mFasL (53.81 +/- 6.9%), and significantly induced the apoptosis of Fas+ Yac-1 cells (56.78 +/- 4.5%), as both were cocultured for 5 h at 1:1 ratio, whereas it is 7.08 +/- 3.4% in control group (P < 0.01). Supernatant from PLFIN-PA317 could also directly induce the apoptosis of Yac-1 within 5 h of incubation. Thus, the culture supernatants from PLFIN-PA317 possessed both infectivity and cytotoxicity of mFasL. PMID- 12539581 TI - Anti-viral activity of hairpin ribozyme directed against HBV core region in vitro. AB - To study the preparation and cleavage of hairpin ribozyme (HpRz) directed against the transcript of HBV core region in vitro, HRz gene designed by computer targeting the transcript of HBV core gene was cloned into the vector p1.5 between 5'-cis-Rz and 3'-cis-Rz. 32P-labeled HpRz transcript proved whether the vector fit for the preparation of hairpin ribozyme. 32P-labeled pKC transcript containing HBV core region as targets-RNA was transcribed by using T7 RNA polymerase and purified by PAGE. Cold HpRz transcript was incubated with 32P labeled target-RNAs under different conditions and radioautographed after denaturing polyacrylamide gel electrophoresis. The results showed that HpRz had the ability of cleavage at 37 degrees C and 12 mmol/L MgCl2 and the design of ribozyme was correct. It is concluded that HpRz prepared in vitro possesses specific catalytic activity, indicating that it is possible for HpRz to intracellularly inhibit the replication of HBV. It may be developed into a nucleic acid drug in the treatment of hepatitis B in the future. PMID- 12539582 TI - [Establishment of hepatocellular carcinoma model by allogenic transplantation ]. PMID- 12539583 TI - The effect of interleukin-6 on the proliferation of prostate cancer cells in vitro and the modulation of this procedure. AB - The role of interleukin-6 (IL-6) in the growth of an androgen-independent prostate cancer cell line (PC-3 m) was defined and the effect of dexamethasone, which was previously shown to modulate IL-6/IL-6 receptor (IL-6R) on this procedure was investigated. By using a pretty sensitive and specific enzyme immunoassay (ELISA), it was found that PC-3 m produced certain IL-6, but there was no difference in IL-6 secretion between the group with or without dexamethasone treatment. It was also found that PC-3 m cells could not be stimulated to grow by exogenous IL-6 (P > 0.05), while it could be inhibited to grow by anti-IL-6 monoclonal antibody and dexamethasone with a dose-dependent fashion. Our observation indicated that IL-6 acted as an autocrine growth factor for PC-3 m, and dexamethasone could inhibit cell proliferation by a mechanism independent of its effect on IL-6 mRNA expression. PMID- 12539584 TI - Expression of angiopoietin-2 gene and its receptor Tie2 in hepatocellular carcinoma. AB - To explore the relationship of angiogenesis-related angiopoietin-2 gene and its receptor Tie2 with angiogenesis and the biology of hepatocellular carcinoma (HCC), angiopoietin-2 gene, Tie2 and CD34 protein expression in 22 resected HCC, 8 cirrhotic and 8 control liver specimens were investigated by in situ hybridization and immunohistochemistry respectively, and the level of angiopoietin-2 and Tie2 expression in HCC were compared in terms of tumor biological parameters. It was found that CD34 was not expressed in control liver, expressed scarcely in cirrhotic liver (17.8 +/- 13.5/HP), but intensively expressed in HCC (86.3 +/- 34.8/HP, P < 0.01). Tie2 receptor was not expressed in controls, expressed at low level in cirrhotic liver (11.3 +/- 8.7/HP), while strongly positive in the microvascular endothelia of HCC (52.4 +/- 16.7/HP, P < 0.01). The level of Tie2 receptor expression in HCC was closely related with tumor diameter, angiogenesis and portal invasion. Angiopoietin-2 gene was not expressed in control liver, expressed mildly in cirrhotic liver (11.2 +/- 9.7/HP), but extensively in tumor zone (36.4 +/- 17.5/HP), the level of angiopoietin-2 expression was closely related with angiogenesis, portal invasion and histological grading of HCC. It is concluded that angiogenesis is increased in HCC; angiopoietin-2/Tie2 expression in human hepatic carcinoma is closely related with angiogenesis, which are probably involved in the HCC angiogenesis regulation, promoting the development and metastasis of human hepatic cancer. PMID- 12539585 TI - The relationship of CyclinD1 and estrogen receptor expression in the process of proliferation and metastasis in breast neoplasm. AB - The role of CyclinD1 and estrogen receptor (ER) in the process of proliferation and metastasis of breast neoplasm and their relationship were studied. The expression levels of CyclinD1 and ER in the tissue samples were detected by using flow cytometry and L SAB immunohistochemistry staining, respectively. The results showed that CyclinD1 and ER expression levels in breast cancer were significantly higher than in benign breast neoplasm (P < 0.05). The CyclinD1 expression levels in stage I was much lower than in stages II, III, IV (P < 0.05). The positive rate of ER was not related with tumor size, lymph node metastasis and TNM stage (P > 0.05), but the CyclinD1 expression level in ER (+) group was significantly higher than in ER (-) group (P < 0.05). It was concluded that CyclinD1 expression level might be obviously related with the proliferation and metastasis of breast neoplasm and ER. PMID- 12539586 TI - A modified method for preparation of adriamycin carried by magnetic albumin microspheres. AB - The targeting of antineoplastic agents to restricted anatomic sites and specific target cells have been challenged clinicians all the time in cancer chemotherapy, which resulted in recent efforts to focus the effects of existing antitumor agents and treatments on tumor cells and spare their effects on normal cells. The drug-carrier complex, adriamycin carried by magnetic albumin microspheres (ADM MAM) was prepared by using our discovered new and modified method. The physical feature of the prepared drug-carrier microspheres was much better than by the traditional method in comparison. The successful preparation of the drug-carrier complex, ADM-MAM, is one of the key steps for our later further researches in the targeted chemotherapy. PMID- 12539587 TI - Ultrasonographic diagnosis of bone tumor of the knee and its clinical implication. AB - In order to evaluate the value of the ultrasonography in the diagnosis of tumor of the knee and its clinical implication, 67 patients with clinically suspected bone tumor of the knee were examined by ultrasound. The ultrasonographic characteristics of different bone tumors were studied and compared with the results of pathologic characters after operation. Ultrasonography can readily visualize the bony destruction and the pathologic change of the periosteum and the soft tissue related to bone tumor. Fifty-two cases of malignant bone tumors and 15 cases of giant cell tumors were diagnosed by ultrasonography. Pathologically, there were 54 cases of malignant bone tumor and 13 cases of giant cell tumor. It was concluded that ultrasonographic examination might be a useful method for the diagnoses of bone tumor of the knee and play an important role in guiding needle biopsy and electing operative method and approach. PMID- 12539588 TI - Hepatitis G viral RNA co-infection in plasma and peripheral blood mononuclear cells in patients with hepatitis C. AB - The incidence of the co-infection of hepatitis G virus (HGV) and hepatitis C virus (HCV) and its clinical implication was investigated and the difference in the positive rate of HGV RNA and HCV RNA between plasma and peripheral blood mononuclear cells (PBMCs) observed. By using reverse transcriptase polymerase chain reaction (RT-PCR) assay, HCV-RNA and HGV-RNA in plasma and PBMCs of 72 patients with hepatitis C was detected. It was showed that HGV RNA was positive in plasma of 11 patients, in PBMCs of 15 patients, and simultaneously in both of plasma and PBMCs of 10 patients with the co-infection rate being 22.2%. Nine patients were both HGV RNA and HCV RNA positive in plasma, 11 patients were both HGV RNA and HCV RNA positive in PBMC, and 6 patients were both HGV RNA and HCV RNA positive in both plasma and PBMC with the positive rate being 12.4%, 15.3% and 8.3% respectively. The positive rate of both HGV RNA and HCV RNA in PBMCs was higher than in plasma. It was concluded that the HGV co-infection rate in the patients with hepatitis C was 22.2%. Simultaneous examination of plasma and PBMC can improve clinically detectable rate. PMID- 12539589 TI - Determination of CD30 expression on peripheral blood T lymphocyte subsets in patients with hemorrhagic fever with renal syndrome by FCM. AB - To determine the CD30 expression on peripheral blood T lymphocyte subsets in patients with hemorrhagic fever with renal syndrome (HFRS) and its clinical implications, double immunofluorescence technique and flow cytometry were used. There was no significant difference among the severe group, mild-moderate group and normal control group in the CD4+CD30- T lymphocyte subset. While the CD4+CD30+ T cells of HFRS patients were increased and the difference between severe group and mild-moderate group or normal control group were very significant (P < 0.01) and the difference between the mild-moderate group and normal control group was also significant (P < 0.05). The CD8+CD30- T cells were increased while the CD8+CD30+ T cells decreased obviously in HFRS patients, and the differences among three groups in both subsets were very significant (P < 0.01). The results showed that the humoral immunity and cellular immunity are overactive in HFRS patients during acute phase. The loss of balance between T lymphocyte subsets may play an important role in the pathophysiology of HFRS and is closely correlated with the severity of the HFRS. PMID- 12539590 TI - Clinical significance of magnetic resonance imaging of bone marrow in patients with leukemia. AB - To investigate the clinical significance of magnetic resonance imaging (MRI) of bone marrow in patients with acute leukemia, the femoral and pelvic marrow were evaluated by using MRI with a T1-weighted spin-echo (SE) method and a short T1 inversion recovery (STIR) technique. Normal bone marrow examination was performed with coronal T1-weighted MRI of pelvis and femurs, and showed persistent red marrow. There was a bright signal of fatty marrow in the femoral epiphyses and apophyses. MRI pattern of bone marrow in the 54 cases of acute leukemia showed abnormal signal patterns of femoral and pelvic marrow: (1) grade I (n = 4), (2) grade II (n = 11), (3) grade III (n = 8), (4) grade IV (n = 17), and (5) graded V (n = 14). Leukemic cells had infiltration onseted by red marrow in adult patients with leukemia. The marrow of femur had infiltration from diaphysis to epiphysis, and to femoral head and greater trochanter. The lower grades (grade IV, V) of leukemic marrow supported the diagnosis of AML in MRI, which achieved higher complete remission. The adult patients with ALL had higher grades (grade I-III) in MRI. Our findings indicated that MRI of femoral marrow is an important tool for accurate diagnosis and management of patients with leukemia that may function as an adjunct to bone marrow aspiration and biopsy. The pattern of MRI in patients with newly diagnosed leukemia predicted the prognosis and CR of leukemia. PMID- 12539591 TI - Effect of pressure on intracardiac backscatter from microbubbles. AB - The backscatter from sonicated albumin microbubbles (Albunex) was analyzed using acoustic densitometry in an in vitro pulsatile heart model to evaluate the effects of pressure on the backscatter from Albunex, and the cardiac cyclic changes of intracardiac backscatter from sonicated albumin microbubbles in 16 healthy persons were analyzed. It was found that the Albunex microbubbles were compressed in systole and decompressed in diastole, causing corresponding changes of backscatter in cardiac cycle. Although the intensities of backscatter in diastole and systole were related to the concentration of microbubbles, the concentration of microbubbles had no effect on the difference of end-diastolic and end-systolic backscatter. The difference of the backscatter was highly correlated with end-systolic pressure (r = 0.96, P = 0.001). In human studies, we also observed same intracardiac cyclic changes of backscatter from sonicated albumin microbubbles. Our study indicates that it is possible to evaluate the intracardiac pressure non-invasively by analyzing the intracardiac backscatter from the microbubbles with acoustic densitometry. PMID- 12539592 TI - [Determination of cytomegalovirus immunoglobulin G and M antibodies by filter paper technique in newborn infants ]. PMID- 12539593 TI - The predictive value of plasma fibronectin concentration on fetal growth retardation at earlier stage of the third trimester. AB - In order to evaluate the predictive value of maternal plasma fibronectin (FN) concentration at 24-34 weeks on fetal intrauterine growth retardation (IUGR), a prospective double-blinded study was performed. The maternal plasma FN concentrations were measured by using a rate nephelometric procedure in the 130 initial normal nulliparous pregnant woman at 24-34 gestational weeks. The outcome of pregnancies and birth weight of their infants were followed up. IUGR was defined as that the birth weight was less than the 10th percentile for gestational age. The receiver operating characteristic curves and predictive values of FN predicting on outcome of pregnancy with IUGR were analyzed. The results showed that: (1) In a cohort of 130 initially normal nulliparous pregnant women, IUGR occurred in 14 cases during the follow-up; (2) The plasma FN levels in the women with IUGR (467.58 +/- 104.43 mg/L) were significantly higher than in the normal control group (299.44 +/- 105.55 mg/L, P < 0.01). However, there was no significant difference in the mean maternal age, gravidity, sampling gestational ages, delivering gestational ages between the two groups (P > 0.05); (3) The areas under ROC curve for predicting the outcome of pregnancy in IUGR was 0.893; (4) At the cut point of 475 mg/L FN level, the sensitivity, specificity, positive predictive value, negative predictive value and Kappa index for predicting the outcomes of pregnancy in IUGR were 57.14%, 95.69%, 61.54%, 94.87%, 0.5455 respectively. It was concluded that the maternal plasma FN might be used as an earlier predictor for screening of IUGR. PMID- 12539594 TI - The study on intramyocardial calcium overload and apoptosis induced by coxsackievirus B3. AB - The isolated cardiac myocytes of rats were immediately infected by cosackievirus B3 (CVB3) to investigate the effects of such procedure on the cell cycle, apoptosis and intracellular ionized calcium (Ca2+ i) of cardiac myocytes. Newborn Balb/c murine cardiac myocytes were cultivated, then infected by CVB3. Intracellular Ca2+ i was measured by flow cytometer. The calcium in the medium for culturing cardiac myocytes was detected by using atom absorb spectrum test. It was found that CVB3 could markedly inhibit the differentiation and proliferation of the infected cardiac myocytes and induce the apoptosis. The intracellular Ca2+ i level in the infected group was significantly higher than in the control group (P < 0.01). The calcium concentration in the medium for culturing cardiac myocytes in the infected group was significantly lower than in the control group (P < 0.05). It was suggested that the apoptosis and intracellular calcium overload of the CVB3-affected cardiac myocytes are likely to play an important role in the pathogenesis of viral myocarditis. PMID- 12539595 TI - Analysis of the factors influencing the therapeutic effects of onychomycosis. AB - In order to improve the curative effect of onychomycosis, the factors influencing the therapeutic effects were investigated. 545 cases including 245 males and 300 females, who were diagnosed both clinically and mycologically, were treated by Intraconazole with intermittent pulse therapy. The therapeutic effects were judged by the following observations regularly and analyzed from the factors as follows: age; growing speed of nails; accompanied diseases; family history; trauma of nails; infection ways of the pathogens; manifestation of the injury; pathogens; duration of the treatment. The results showed that the recovery rate was higher in younger patients (P < 0.01) with a quicker recovery rate (P < 0.001), and a lower recurrent rate (P < 0.01), as well as in those with quicker growing speed of new-born nail. Also the patients with WSO and DLSO manifestation had a higher recovery rate. The patients with onychomycosis caused by T. rubrum had a higher recovery rate (P < 0.01 to 0.001) no matter whether to prolong the treatment duration. The patients with diabetes mellitus or hyperhidrosis, as well as with positive family history or basic nail diseases such as trauma and paronychia, had a lower recovery rate and the curative effects were not satisfactory. It was concluded that although the single and some DLSO manifestation nail injury could be cured by internal and external treatments with the help of removing the sick nail and the duration of the treatment could be shortened. The treatment duration should be prolonged in order to increase the curative effects and decrease the recurrence under such conditions as following: old patients above 60 years; patients with low-growing-speed new-borne nails; patients with thumb and big toel injury and ingrowing nail; patients with diabetes mellitus, hyperhidrosis or Renauld's phenomenon; patients with nail trauma before or during the treatment; patients with PSO or TDO manifestation; patients with onychomycosis caused by Candida or Aspergillus; patients with abnormal new-borne nails of abnormal color, coarse surface or abnormal thickness. PMID- 12539596 TI - Expression of P53 during lens epithelial cell apoptosis induced by ultraviolet. AB - The apoptosis of lens epithelial cells (LECs) induced by ultraviolet and the expression of P53 were investigated. Wistar rats received 100 mW/m2 ultraviolet irradiation (UVR) (lambda = 280 nm-315 nm) for 15 min. One, 6, 24 h after irradiation the lens capsules were dissected. The percentages of apoptotic cells were evaluated by the TdT-dUTP terminal nick-end labeling (TUNEL) technique and the expression of P53 was detected by using immunohistochemical assay. The results showed that the percentages of TUNEL-positive nuclei at 24 h after irradiation was significantly higher than in the control group and those 1 h, 6 h after irradiation. The percentages of P53-positive cells at 6 h, 24 h after irradiation were significantly higher than in the control group and those 1 h after irradiation. It was concluded that UVR could induce the apoptosis of lens epithelial cell. The expression of P53 might be responsible for the apoptosis of lens epithelial cells. PMID- 12539624 TI - [Expression of VEGF-C and its correlation with cervical lymph nodes metastasis of oral cancers]. AB - OBJECTIVE: Vascular endothelial growth factor C (VEGF-C) is known to be capable of inducing cell proliferation of lymphatic endothelia and development of lymphatic vessels. Thus, in the current study the expression of VEGF-C was investigated to understand the mechanism of lymphatic metastasis of oral cancer. METHODS: VEGF-C protein expression in benign and malignant oral lesions was investigated with an immunohistochemical staining assay, followed by light microscopic examination and image analysis. RESULTS: Immunohistostaining of VEGF C was observed in normal, benign and malignant epithelia, as well as in some lymphatic vessels. The VEGF-C positive rate was significantly higher in squamous carcinomas than in normal tissues and benign lesions. It was also noted that in squamous carcinomas the expression level of VEGF-C correlated well with pathological grades and stages of lymph node metastasis, but not with clinical grades. CONCLUSION: These findings suggest that VEGF-C may promote lymphatic metastasis of oral cancer, and it is possible to be used as a predictor for the lymphatic metastasis and prognosis of oral cancer. PMID- 12539625 TI - [Ultrastructural and Ca/P changes of the regenerated bone after mandibular distraction in goats]. AB - OBJECTIVE: The aim of this study was to examine the ultra structural and the changes of Ca/P elements of the new bone resulting from mandibular distraction osteogenesis. METHODS: Bilateral mandibular osteotomies were performed in six goats. The mandibles in all animals were lengthened 10 mm using a custom made distracter. Each 2 goats were sacrificed respectively at 2, 4 and 8 weeks after completion of distraction. The distracted callus were harvested and processed for SEM evaluation and Ca/P ratio analysis. RESULTS: The newly generated bone in distraction gap was confirmed by SEM examination, and the contents of Ca, P were found to be elevated with the maturity of distracted callus. CONCLUSION: This study suggested that newly formed bone by mandibular distraction osteogenesis in goats could be remodeled to normal bone. PMID- 12539626 TI - [Buccal swab: a convenient source of DNA for analysis of IL-1 gene polymorphisms]. AB - OBJECTIVE: PCR-RFLP based techniques have become standard procedures for gene polymorphism screening. Peripheral venous blood is currently the most commonly employed source of DNA for human genome analysis. However, it has many practical disadvantage and inherent limitations to use blood as DNA source. Blood sampling is invasive, painful and involves a potential risk of contamination with hepatitis. The classic procedure to extract genomic DNA from whole blood is phenol/chloroform technique, which is relative complicated and time consuming. Therefore, this study was performed to try an alternative instead of using blood as DNA source for gene polymorphism analysis. METHODS: Four methods were employed to obtain DNA from the same subject including DNA from venous blood through phenol/chloroform extraction, DNA from dried blood spot through Chelex 100 technique, DNA from buccal swab through Chelex 100 technique and the dried blood spot directly as DNA template for PCR. Then, these various forms of DNA were used in PCR RFLP procedure to analyze IL-1 gene polymorphisms, and their specificity and sensitivity were evaluated. RESULTS: Our results indicate that both the buccal swab and the blood based assays reached complete concordance in typing the IL-1 gene polymorphisms, while the Chelex 100 procedure for extracting DNA from buccal swab is much simpler and more rapid. It is non-invasive to get buccal swab. The amount of DNA obtained through one buccal swab is 63.8 micrograms +/- 18.7 micrograms, which is enough for 10 PCR reactions. CONCLUSION: Buccal swab appears to be an excellent source of DNA for detection of polymorphisms of human IL-1 gene. PMID- 12539627 TI - [Relationship between the expression of Bax and apoptosis induced by hyperthermia in BcaCD885 cells]. AB - OBJECTIVE: This study aimed to investigate the role of Bax gene in apoptosis induced by hyperthermia in BcaCD885 cells. METHODS: After inducing apoptosis through hyperthermia, the expression of Bax protein and mRNA were examined with FCM antibody analysis and RT-PCR technique respectively. RESULTS: Up regulation of Bax protein was noted after hyperthermia. The correlation coefficient r between the apoptotic rate and the expression Bax protein was 0.9240. The expression of Bax mRNA was also raised after hyperthermia. CONCLUSION: Hyperthermia acting as an external signal may up-regulates expression of Bax gene and then induces apoptosis in BcaCD885 cells. PMID- 12539628 TI - [Study on maxillofacial impact injury associated with brain injury]. AB - OBJECTIVE: The purpose was to study the mechanism and the characteristics of brain damage associated with maxillofacial injury under the impacting load of middle face. METHODS: Eighteen rabbits were subjected to impacting on left middle face with pneumatic impact device, which were horizontally located on the ground. The impact velocities respectively were 4.77 m/s +/- 0.53 m/s (group A), 9.16 m/s +/- 0.65 m/s (group B) and 13.95 m/s +/- 0.67 m/s (group C). The acceleration of head and stress of encephalic was monitored during the impact period. The pathological characteristics of correlative tissues were examined in detail 6 hours after impacting. The contents of LPO, SOD and NO in blood and water in brain were measured at the same time. RESULTS: The contents of LPO and SOD in blood significantly increased along with the increasing of impact velocity (P < 0.05). The same pattern was observed in the changing of water content of brain (P < 0.05). But the contents of NO in blood reduced when the impact velocity increased (P < 0.05). The animals of group A showed single fracture of middle face bone, while Group B showed multiple fractures of middle face bone. Group C represented severe maxillofacial injury associated with brain damage. The pathological damage of brain became more serious along with the increase of impact velocity. The peak value of head acceleration and encephalic stress of group C were higher than that of group A and group B (P < 0.05). CONCLUSION: The responses of head acceleration and encephalic stress resulting from middle face impacting might play a key role in brain injury associated with maxillofacial injury. PMID- 12539629 TI - [Comparison of the effect of fluoride and non-fluoride dentifrices on enamel demineralization and remineralization in vitro]. AB - OBJECTIVE: The purpose of the experiment is to investigate the effect of fluoride dentifrices in China on the intact and artificial caries-like bovine enamels. METHODS: The artificial caries-like lesions were created by dipping the teeth into an acid buffered solution. Both the bovine intact enamels and the artificial caries-like enamels were assigned tooth brushing 20 times every half an hour with 3 kinds of fluoride dentifrices and one kind of non-fluoride dentifrices respectively. The amount of calcium dissolved by lactic acid was determined as the susceptibility of demineralization by atomic adsorption spectrometry (AAS). RESULTS: The group treated with fluoride dentifrice showed significantly less demineralization than the non-fluoride group (P < 0.02). The effect of inhibition of acid solubility in artificial caries-like enamel after tooth brushing is significantly greater than that in intact enamel. CONCLUSION: Fluoride dentifrices in this experiment can prevent both intact enamel and artificial caries-like enamel from acid solubility after tooth brushing, and it seems that the effect of the fluoride dentifrices on the redemineralization is greater than the reduction of enamel solubility. PMID- 12539630 TI - [The relative factors of protrusive balanced occlusion and the relationship among them]. AB - OBJECTIVE: The experiment was designed to study the condition for achieving protrusive balanced occlusion and the factors involved in occlusal equilibration. METHODS: For achieving protrusive balanced occlusion, the posterior teeth of mandible always contacted with the posterior teeth of maxilla in the process of protrusive occlusive movement. The angle between the line of movement track of mandible cusp in the process of protrusive occlusive movement and occlusion plane is called cusp movement inclination. A formula of calculation was obtained by using mathematics track and the conversion of coordinate axis. RESULTS: The focus of protrusive balanced occlusion is relationship between the track of mandibular cusp movement and working incline of maxillary cusp. The first condition achieving protrusive balanced occlusion is that cusp movement inclination is equal to cusp working inclination of maxilla. The factors affecting cusp movement inclination are condyle path inclination; incisal path inclination and teeth coordinate. The factors affecting working inclination of cusp are cusp inclination and long axis obliquity. CONCLUSION: Cusp movement inclination is the basic and the most important concept in protrusive balanced occlusion. PMID- 12539631 TI - [Preliminary study of closing cleft of soft palate at an early age for complete cleft palate]. AB - OBJECTIVE: The aim is to introduce the method closing the soft palate at an early age in order to produce better speech and lessen interruption to the development of maxilla. METHODS: Clinical data of 37 cases with treated cleft were preliminarily analyzed. Some dental casts were measured. RESULTS: 1. No complications of anaesthesia and surgery. Except 2 cases whose soft palates were split after surgery were observed as they were coughing. 2. A reduction in the width of the hard palate cleft was noticed. CONCLUSION: Closing the soft palate at an early age is possible and safe. It shows that the Langebeck method could be used to close the hard palate of patients at an early age. PMID- 12539632 TI - [The study of operative design in unilateral cleft lip-repair between individual procedural method as compared to rotation advancement and triangular flap method]. AB - OBJECTIVE: To compare the relationship and differences between individual operative procedural design with the Millard and Tennison methods in cleft lip repair. METHODS: The author specifically analyzed and compared the end point of incision on noncleft side of unilateral cleft lip in three kind of operative respectively. In addition, the operative design was discussed and found out to be more reasonable. RESULTS: The individual operative procedural design have wider indication and adaptable to change and function than Millard and Tennison methods alone from the geometrical configuration point of view. CONCLUSION: Therefore, the author conclude that the individual operative procedural design is practical and applicable than other operative design in cleft lip repair. PMID- 12539633 TI - [Cephalometric analysis of the patients with bimaxillary protrusion in south China]. AB - OBJECTIVE: The purpose of this article is to characterize the teeth, jaws and soft tissues of the patients with bimaxillary protrusion in south China. METHODS: The results of cephalometric analysis of 60 patients with bimaxillary protrusion were compared with the normal occlusions in Beijing. RESULTS: Compared with the normals in Beijing, the mandibles of the subjects were positioned more backward and rotated inferior-posteriorily. The incisors inclined extremely to the labial side and the lower lip was thicker. The mental thickness was less than the normal, and the mental contour was not clear. The middle face was eminent. CONCLUSION: The results suggest it is of critical importance to manage to adjust the position of jaws, to control the molar anchorage strictly, and to achieve the maximal incisor retraction, so as to benefit the improvement of facial features. PMID- 12539634 TI - [Clinical effects of metronidazole-ofloxacin gel as an adjunct to conventional therapy for periodontitis]. AB - OBJECTIVE: This study was designed to evaluate the clinical effects of metronidazole ofloxacin gel (MO gel) as an adjunct to conventional therapy for periodontitis. METHODS: 13 patients with adult periodontitis were selected and each of them had two or more sites with probing depth > or = 4 mm, so totally 35 teeth were included in this study. After conventional therapy, the subjects were randomly divided into three groups: (1) experimental group application with MO gel, (2) standard group application with metronidazole stitus, and (3) control group application with placebo gel. Local application was performed once a week for 12 weeks. Clinical parameters including PD, PAL, GI, BOP, PLI was recorded after 6 weeks and 12 weeks respectively. RESULTS: All clinical parameters from group of gel and group of metronidazole stilus were better than that from control group, while no significant difference was observed between MO gel group and metronidazole stilus group. CONCLUSION: It indicates local application with MO gel treatment as an adjunct to conventional therapy may lead to a more advantageous outcome for periodontitis. PMID- 12539635 TI - [The optimal concentration of drinking water in eastern Guangdong]. AB - OBJECTIVE: The objectives were to explore the optimal concentration of drinking water in Eastern Guangdong and to provide information for the health administrative department to set up the criteria for fluoride concentration. METHODS: Totally 1237 9-17 and 35-44 years old residents in Eastern Guangdong who drinking water containing 0.1-2.5 mg/L fluoride were recruited to investigate the occurrences of dental caries and dental fluorosis. Their background information and perceived appearances of teeth were also recorded. RESULTS: The major factor influencing dental fluorosis was fluoride concentration in drinking water. Dental caries decreased when the fluoride concentration increased but dental fluorosis also increased at the same time. It was showed by calculation that the Dental Fluorosis Index would be 0.4 and 0.6 when the fluoride concentration was 0.57 mg/L and 0.7 mg/L respectively. The acceptation to the severity of dental fluorosis among the study subjects was in accordance with the Dean's criteria. CONCLUSION: It suggests that the optimal fluoride concentration in Eastern Guangdong is 0.57 mg/L and should not exceed the upper limit of 0.7 mg/L. PMID- 12539636 TI - [Study of bacteriology on local application of Tinidazole stilus in treatment for periodontitis]. AB - OBJECTIVE: The study was designed to evaluate the antiseptic efficacy of Tinidazole stilus against periodontal pathogen. METHODS: A total of 41 patients with periodontitis were included in this study. 26 of them were treated by Tindazole stilus and other 15 cases were treated by metronidazole stilus as controls. The prevalence of subgingival bacteria from both group were examined. RESULTS: The eliminate rates for Black pigmented anaerobic rod (Porphyromonas gingivalis, Prevotella intermedius, Prevotella melaninogenicus), Fusobacteriums, Prevotella oris, Prevotella oralis, Eubacterium and Actinomyces were 95.8% 94.1%, 83.3% and 76.5%. The anti-bacterial efficiency of Tinidazole against periodontal pathogen was significantly higher than that of metronidazole (P < 0.05). CONCLUSION: It appears to be effective to use Tinidazole stilus as local application against periodontal pathogens. PMID- 12539637 TI - [The influences upon the passive tensile of the masticatory muscles and ligaments by Herbst appliance under various bite reconstruction--a three dimensional finite element analysis]. AB - OBJECTIVE: This study aimed to provide some biomechanical references for the clinical use and improvement of Herbst appliance. METHODS: The three-dimensional model of the 'Temporomandibular joint mandible Herbst appliance system' was set up by SUPER SAP software (version 9.3). On this model, the passive tensile in the masticatory muscles and ligaments were analyzed under various bit reconstruction designed according to specified advanced displacement and vertical bite opening. RESULTS: When Herbst appliance drove the mandible forward, there was not any tensile in the medial, lateral pterygoid and the collateral ligament, while the temporalis, the deep paret of masseter, the stylomandibular ligament, and the sphenomandibular ligament were passively drawn. Under various bite reconstruction, the passive tensile in the medial temporalis, the posterior temporalis, the stylomandibular ligament and the sphenomandibular ligament increased with the amount of the advancement of the mandible; The passive tensile in the posterior temporalis and the deep part of masseter increased with the amount of vertical bite opening of the mandible. CONCLUSION: The StL, SpL, PT and AT played an important role in functional reconstruction of mandible by Herbst. All five group of bite reconstruction (3-7 mm advancement, 4-2 mm vertical bite opening of the mandible) designed by this study can be selected in clinic according to the patient's capability of adaptation, the extent of malocclusion and the potential and direction of growth. PMID- 12539638 TI - [The influences of the stress distribution on the condylar cartilage surface by Herbst appliance under various bite reconstruction--a three dimensional finite element analysis]. AB - OBJECTIVE: The distribution of stress on the surface of condylar cartilage was investigated. METHODS: Three-dimensional model of the 'Temporomandibular joint mandible Herbst appliance system' was set up by SUPER SAP software (version 9.3). On this model, various bite reconstruction was simulated according to specified advanced displacement and vertical bite opening. The distribution of maximum and minimum principal stress on the surface of condylar cartilage were computerized and analyzed. RESULTS: When Herbst appliance drove the mandible forward, the anterior condyle surface was compressed while the posterior surface was drawn. The trend of stress on the same point on the condyle surface was consistent in various reconstruction conditions, but the trend of stress on various point were different in same reconstruction conditions. CONCLUSION: All five groups of bite reconstruction (3-7 mm advancement, 4-2 mm vertical bite opening of the mandible) designed by this study can be selected in clinic according to the patient's capability of adaptation, the extent of malocclusion and the potential and direction of growth. PMID- 12539639 TI - [Research and establishment of the computerized orthodontic and orthopedic prediction system]. AB - OBJECTIVE: The aim of this study was to develop a computerized orthopedics and orthodontics interactive prediction system and verify its reliability. METHODS: 24 cases of Angel II division I patients were selected in the study. All of them had been treated by activator or Frankel functional appliance, and had the satisfied therapy result. Both pre and post-treatment cephalogram of them were used verifying veracity of therapy prediction. RESULTS: All of cephalometric measure items include 11 angle measure items and 14 distance measure items. There were 9 coincidence indicators in angle measure items and 7 in distance measure items. CONCLUSION: The results indicate it is liable and effective to use the computerized orthopedics and orthodontics system for prediction. PMID- 12539640 TI - [Establishment and research of the computerized profile and photo prediction system]. AB - OBJECTIVE: The study aims to develop a lateral oral facial profile and photo prediction system and then to verify its reliability. METHODS: 24 paired lateral cepholagrams in both pre- and post-treatment were selected and analyzed by three soft tissue cephalometric analysis methods in this study. All cases were of the angel II division I malocclusion, and had been successfully treated with the orthopedics therapy by activator or Frankel appliance. Soft tissue profile prediction had been automatically completed, following the hard tissue interactive orthopedics prediction approach in the pretreatment lateral cephalogram, and the predictive results were compared to the same cepholametric analysis in post-treatment cephalogram. RESULTS: There are 19 coincidence indicators in 30 measurements of three soft tissue cephalometric analysis, and 7 in 9 angel measurements, 10 in 18 instance measurements, 2 in 3 instance ratio measurements is consistency, respectively. CONCLUSION: The results indicate that the lateral oral facial profile and photo prediction system is effective in profile prediction. PMID- 12539641 TI - [Shear strength of bond of the dentin to GI-II glass-infiltrated alumia ceramic agter varying modes of surface conditioning]. AB - OBJECTIVE: The adequate surface conditioning method for GI-II glass infiltrated alumia ceramic was investigated. METHODS: The test specimens were divided into 3 groups (A. HF + silicon, B. sandblasting + silicone and C. thermal silicoating + silicone). Shear bonding strengths of porcelain bonded to dentin were tested. RESULTS: The bond strength of group A and B slightly increased after one day but decreased sharply after 30 days. However, no statistical significance was observed. The group C showed a pronounced enlargement of bond strength after one day and had a durable bond after 30 days' thermal cycling. It is statistically significant when group C was compared with group A and B. CONCLUSION: It can be inferred that silicoating method be a better way for glass infiltrated alumia ceramic bonding. PMID- 12539642 TI - [The function of benzol peroxide in the induction of Syrian golden hamster tongue carcinoma by chemical carcinogen]. AB - OBJECTIVE: To testify the function of promoter(benzol peroxide) in the induced tongue carcinogenesis of Syrian golden hamster as well as the function of carcinogen (DMBA). METHODS: In this study, group 1: 20% benzoyl peroxide in acetone solution was brushed to the right tongue border of Syrian golden hamster, followed by 0.5% 7, 12-dimethybenzanthrancene(DMBA) in acetone, twice a week, lasting 20 weeks. Group 2: 0.5% 7,12-dimethybenzanthrancene(DMBA) in acetone was painted on the right tongue border of golden Syrian hamster in the manner of group 1. Group 3: 20% benzoyl peroxide in acetone solution was used with mentioned methods. RESULTS: Group 2: showed 90% (27/30) carcinogenesis-rate in tongue of Syrian golden hamster. While it was 100% (30/30) of induced tongue carcinoma and 3/30 lymph node metastasis was found in group 1. No tongue carcinogenesis was found in group 3. The former two groups had the manifestations of hyperplasia of epithelium cell, carcinoma in situ, invasive and cervical lymph node metastatic carcinoma in the process of experiment, but it was 2 weeks shorter in group 1 than that of group 2. Both groups had the same features under eye and light microscope. The samples were high-differentiated squamous cell carcinoma histologically through HE stain. CONCLUSION: This study indicated that benzoyl peroxide could increase carcinogenesis of golden hamster tongue which induced by DMBA. Consideration of promoting agent should be taken in carcinogenesis as well as carcinogen. PMID- 12539643 TI - [A surface reacted layer study of titanium-zirconium alloy after dental casting]. AB - OBJECTIVE: To investigate the influence of the mold temperature on the surface reacted layer of Ti-Zr alloy castings. METHODS: Ti-Zr alloy was casted into a mold which was made of a zircon (ZrO2.SiO2) for inner coating and a phosphate bonded material for outer investing with a casting machine (China) designed as vacuum, pressure and centrifuge. At three mold temperatures (room temperature, 300 degrees C, 600 degrees C) the Ti-Zr alloy was casted separately. The surface roughness of the castings was calculated by instrument of smooth finish (China). From the surface to the inner part the Knoop hardness and thickness in reacted layer of Ti-Zr alloy casting was measured. The structure of the surface reacted layer was analysed by SEM. Elemental analyses of the interfacial zone of the casting was made by element line scanning observation. RESULTS: The surface roughness of the castings was increased significantly with the mold temperature increasing. At a higher mold temperature the Knoop hardness of the reactive layer was increased. At the three mold temperature the outmost surface was very hard, and microhardness data decreased rapidly where they reached constant values. The thickness was about 85 microns for castings at room temperature and 300 degrees C, 105 microns for castings at 600 degrees C. From the SEM micrograph of the Ti Zr alloy casting, the surface reacted layer could be divided into three different layers. The first layer was called non-structure layer, which thickness was about 10 microns for room temperature group, 20 microns for 300 degrees C and 25 microns for 600 degrees C. The second layer was characterized by coarse-grained acicular crystal, which thickness was about 50 microns for three mold temperatures. The third layer was Ti-Zr alloy. The element line scanning showed non-structure layer with higher level of element of O, Al, Si and Zr, The higher the mold temperature during casting, the deeper the Si permeating and in the second layer the element Si could also be found. CONCLUSION: The mold temperature is one of the major factors influencing to casting quality. In order to reduce the surface reacted layer of Ti-Zr alloy castings, the lower mold temperature and the investment without Si should be chosen. PMID- 12539644 TI - [The investigation of the testing method on fracture toughness of dental adhesive resins]. AB - OBJECTIVE: The testing method for mode I fracture toughness (GIC) of dental adhesive resins was investigated. METHODS: The dental adhesive resins were placed between two Co-Cr alloy plates to make up test specimen, so test specimen had a metal/adhesive resin/metal construction. The category of adhesive resins, the dimensions of test specimen and testing speed were used as the variants. The testing method for mode I fracture toughness (GIC) was investigated by using double cantilever beam (DCB) testing. The correlation between the GIC and each factor was analysed by regression analysis. RESULTS: In all test specimens, it was found that the values of fracture toughness (GIC, GIR) at the process of crack propagation were nearly constant. Therefore, it was found that fracture toughness (GIC) at maximal load might only be measured to evaluate an adhesive resin. On the values of GIC, there was no effect from the dimensions of the test specimens, width (b = 3.5 mm), thickness (2 hm = 2.4 mm), initial crack length (ao = 10-19 mm) and testing speed(Vc = 0.1 mm/min, 0.2 mm/min). But, it was found that the fracture toughness was influenced by thickness of adhesive layer (hr). In case of SB, the value of GIC had a tendency to increase as the adhesive layer became thicker, and then the crack propagating area observed as cohesive failure became remarkably rougher. In case of PV, the values for hr = 100 and 200 microns were similar, however, that for hr = 20 microns somewhat decreased resulting in an interfacial failure. By the way, cohesive failures of hr = 100 and 200 microns were observed as similar flat surfaces. CONCLUSION: Fracture toughness was not influenced very much by the dimensions of test specimen, but was influenced by thickness of adhesive layer. Thus, it was recognized that this testing method was more effective for evaluating the adhesive resin. PMID- 12539645 TI - [The neutralizing effect of histidine-rich-polypeptides on LPS from suspected periodontal pathogens]. AB - OBJECTIVE: Human parotid histidine-rich-polypeptides (HRPs) are a family of lowmolecular-weight, cationic polypeptides. HRP-1, HRP-3 and HRP-5 comprise 85% 90% of the total HRPs and are called major HRPs. There were many researches indicating the antimicrobial activities of HRPs. Recently, HRPs were reported to have an inhibitory action on the lipopolysaccharide (LPS) of E. coli, with HRP-5 being the most effective one among the three major HRPs. Since the LPS of oral gram-negative bacteria is thought to be one of the important etiological factors during the development of periodontal diseases, our experiment was aimed to investigate the neutralizing effect of human parotid HRP-5 on the LPS of anaerobic suspected periodontal pathogens, which have different chemical structures and biological activities compared with LPS of aerobic E. coli. METHODS: By using of preparative acid urea polyacrylamide gel electrophoresis (AU PAGE), HRP-5 was purified from parotid saliva collected from healthy adults. Two stains of suspected periodontal pathogens, Porphyromonas gingivalis (P.g, 47-A) and Fusobacterium nuceatum (F. n, separated from subgingival plaque of a patient with adult periodontitis) were mass cultured. After harvested in the midlogarithmic phase, they were washed and lyophilized. The LPS of dried bacteria were extracted by the modified Westphal hot phenol-water procedures and purified by the enzyme digestion plus ultracentrifugation. Limulus test was applied to test the neutralizing effect of HRP-5 on the LPS-induced gelation of Limulus amoebocyte lysate. In brief, the standard LPS of E. coli, or extracted LPS of P. g or F. n, was preincubated with HRP-5 in a tube separately in room temperature for 10 minutes. Then the reagent of Limulus amoebocyte lysate was added in all the tubes, continued the incubation in 37 degrees C for one hour. After that, the gelation level of every tube was observed. RESULTS: LPS extracted from P. g and F. n both showed good purity and strong activities to induce gelation of Limulus amoebocyte lysate. The gelation induced by LPS (1 ng/ml) of these two anaerobic suspected periodontal pathogens were weakly inhibited by HRP-5 (10 micrograms/ml), similar to that observed with standard LPS of E. coli. To get a complete neutralizing effect on LPS, it may be important to increase the concentration of HRP-5. CONCLUSION: It appeared that HRPs could neutralize the endotoxic properties of LPS of suspected periodontal pathogens, therefore may contribute to periodontal health. The present investigation further confirmed that HRPs are important components of the host non-immune defense system. PMID- 12539646 TI - [The experimental study on inside-out vein graft for repairing inferior alveolar nerve defect in the rabbit]. AB - OBJECTIVE: This study describes a modification of the vein graft technique inside out vein graft, creates an autogenous vein conduct with the collagen-rich and Schwann cells exposed to regenerating axons. To compare regeneration of inferior alveolar nerves through two conduits: inside-out vein graft and the standard vein graft. METHODS: 18 adult white rabbits were selected and vena facialises of both sides were harvested. Turned the right vena facialis into inside-out vein and used to bridge an 8 mm defect created in the right inferior alveolar nerve. An 8 mm defect of the left inferior alveolar nerve was bridged by the standard vein graft. After rabbits sacrificed at 8, 14 and 20 weeks, the regenerating nerve cables were harvested from the conduits and performed histologic study. At 20 weeks, performed also electrophysiologic studies and statistical analysis of conductive velocities and histological results. RESULTS: Regenerating nerves connected with proximal and distal nerve stumps on both sides postoperation 8 weeks, a number of regenerating nerve fibers and thickness of axon were gradually increasing. At 20 weeks regenerating nerve on the inside-out vein graft side showed superior function results--faster conduction velocities (P < 0.05) and distinct histological results--much more axon counts (P < 0.05) and thicker mylin sheaths (P < 0.05) compared with contralateral standard vein graft. CONCLUSION: The adventitial surface of vein promotes nerve regeneration by an environment with collagen-rich and Schwann cells. The inside-out vein graft is an original way to repair inferior alveolar nerve. PMID- 12539647 TI - [Analysis of the expression of BMP-2,3,4,5 in nerve sheath tumors of maxillofacial region]. AB - OBJECTIVE: The aim of present study is to elucidate the relationship between the members of BMP family and the neoplasms of peripheral nerve. METHODS: Eight cases of schwannoma and three cases of malignant schwannoma were collected and all samples were fixed with 10% neutral formalin and embedded in paraffin wax. Five microns thickness sections were prepared for staining with haematoxylin and eosin for histopathological assessment and processed for in situ hybridization for assessing the mRNA transcriptions of BMP-2,3,4,5. Sections of osteosarcoma that previously demonstrated consistent expression of BMP-2 mRNA were included in each batch of staining to serve as positive control. RESULTS: The results showed that all of the neoplastic lesions of nerve sheath showed a consistent expression of BMP-2,3,4,5. The positive signals of benign lesions were obviously increased in the palisading area or the area of accumulated cells. There was no detectable staining in the matrix of tumour and collagenous fiber. The staining intensity was relatively and slightly weak in cells of Antoni B tissue when compared with Antoni A tissue. Nevertheless, the tumour cells in Antoni B tissue also showed BMP-2,3,4 and 5 positively staining. The fibroblasts of schwannoma were negatively stained. The malignant lesions showed higher expression signals of BMPs than benign lesions. CONCLUSION: The results can be concluded that the selected members of BMPs may be involved in the process of growth and progression of the neoplasms of peripheral nerve. PMID- 12539648 TI - [Changes of cultured human vein endothelial cells(HVEC) in the presence of therapeutic agents of cavernous hemangioma]. AB - OBJECTIVE: To observe the damages of cultured human vein endothelial cells treated by various drugs. METHODS: With the identification of EC, damages of which treated by drugs with different concentration were measured with MTT assay respectively, dramatic changes in the morphology of test cells observed under light and electron microscope. RESULTS: 1. According to the different inhibition on proliferation of HVEC, sodium morrhuate exhibited most cytostatic effect for HVEC, and combination of pinyangmycin and hormone produced more significant arrests of growth of HVEC than pinyangmycin used singly. 2. Positive correlation were found between cell contraction and concentration of varied drugs with the absence of structure of junctional integrity. 3. Corresponding pathological changes occurred on HVEC with MTT assay. 4. As the damages of HVEC occurred, therapeutic agents may directly injured substratum of cavernous hemangiomas through enlarged gaps between cells subsequently. CONCLUSION: Evaluation and screening of sclerosing agents for cavernous hemangioma may be feasibly demonstrated with affects on cultured HVEC by different agents. PMID- 12539649 TI - [Distribution of IL-1 beta in periodontium of experimental osteoporosis rats during orthodontic tooth movement]. AB - OBJECTIVE: The purpose of this research is to study the expression change of IL-1 beta induced by forces in the periodontium of osteoporosis rats. METHODS: Eighteen SD rats were randomly divided into three groups: the normal controlled, the normal orthodontically controlled and the experimental group of osteoporosis. Six weeks after ovariectomy, the first molars were moved mesially by 50 g force created from elastic-thread for one week in the force loading groups. Then the rats were killed, and the 5 microns-thick mesiodistal sections of maxillary teeth, including the roots and periodontium were made. IL-1 beta were localized immunohistochemically in periodontium, and semiquantitative analysis of cellular staining intensity was done by microphotometry. RESULTS: The IL-1 beta of periodontal ligament cells without loading force were mildly stained in the controlled group, but the staining intensity increased significantly in the periodontium of orthodontically moved teeth, and it showed more strengthened staining in the periodontium of force-loading teeth in osteoporosis rats. CONCLUSION: The results demonstrate that periodontal ligament cells respond to mechanical force by increased production of IL-1 beta, and this can be enhanced by osteoporosis. IL-1 beta is an important cytokine in the process of orthodontically periodontal remodeling. PMID- 12539650 TI - [Treatment of hemangioma in oral and maxillofacial region with pingyangmycin injection]. AB - OBJECTIVE: To investigate the indication and result in hemangioma in oral and maxillofacial region with Pingyangmycin injection. METHODS: 1282 patients (554 males and 728 females) with hemangioma in oral and maxillofacial region have been treated in our hospital during the 10-year period from May 1990 to March 1999. The patients range between 0.5-84 years old. Lip, cheek and tongue were usually involved. 1211 patients were treated with injection of Pingyangmycin(about 0.89 mg/ml) and dexamethasone (about 0.55 mg/ml), 1 time/1-2 weeks. RESULTS: The curative rates of cavernous hemangioma and strawberry hemangioma were 93.87% and 84.90%, but that of plexiform hemangioma was 22.22% and those of portwine stain and central hemangioma of the jaws were 0%, respectively. Serious complications associated with Pingyangmycin injection, such as pulmonary fibrosis were not seen. CONCLUSION: Injection of Pingyangmycin is selective primary method for cavernous and strawberry hemangioma treatment. PMID- 12539651 TI - [Analysis of maxillary dental-arch dimensions in adult unoperated cleft palate patients]. AB - OBJECTIVE: Analysis of maxillary dental-arch development in adult unoperated cleft palate patients. METHODS: Dental casts was studied in 20 cases of adult unoperated cleft palate (UCP) patients, and 20 cases of normal adults were as a control group. RESULTS: The UCP group showed smaller dental-arch widths than a control group in the canine region (P < 0.05) and these was no significant difference on upper-arch depth and posterior dental arch widths between the UCP group and a control group (P > 0.05). CONCLUSION: In the UCP group major deformity were in incisor and canine region, and smaller interfere to posterior dental-arch widths and depth. Operation for cleft lip can interfere maxillary dental-arch development. PMID- 12539652 TI - [Simultaneous primary palate repair and alveolar bone grafting in unoperated cleft palate patients over 8 years old]. AB - OBJECTIVE: To analyze the applicability and results of simultaneous primary palate repair and alveolar bone grafting in unoperated cleft lip and palate patients over 8 years old. METHODS: A retrospective study was performed in a group of unoperated cleft palate patients who received simultaneous primary palate repair and alveolar bone grafting. Between December 1990 and March 1998, a consecutive of 38 complete unilateral cleft lip and palate patients were treated by the procedures of simultaneous primary cleft palate repair and alveolar bone grafting at the Peking University of Cleft Lip and Palate Treatment Center. All the patients had their lip repaired before they were admitted. The age range was 8 to 24 years, with the average of 14.7 years old. The duration of operation as well as the blood loss during the operation was recorded, and compared with those patients who only received alveolar bone grafting. All the patients have been followed up for at least twelve months, and the results of bone grafting were evaluated according to radiographs. RESULTS: All the operations were successful, and the wound healed well. Compared with simple alveolar bone grafting, simultaneous primary palate repair and alveolar bone grafting prolonged the operation time to an average of 37 minutes. The procedure of simultaneous primary palate repair and alveolar bone grafting did not prolong the operating time compared with simple alveolar bone grafting and no blood transfusion due to bone grafting was needed. All the wounds both in the grafted area and donor site healed uneventfully. No major complications occurred during or shortly after the operation. The overall clinical successful rate of alveolar bone grafting in this group of patients was 89.5%. CONCLUSION: Simultaneous primary palate repair and alveolar bone grafting are safe and applicable procedure for unoperated cleft palate patients, and this procedure should be performed in those unoperated cleft palate patients above 8 years old. PMID- 12539653 TI - [The design, functions and clinic applications of lip bumper]. AB - OBJECTIVE: This paper introduced the principle of design, methods of fabrication, indications and functions of lip bumper, and advocated the application of lip bumper in orthodontic clinic. METHODS: Two typical cases treated with lip bumper were introduced. Some notices and experiences of applying lip bumpers were summed up. RESULTS: There were following functions of lip bumper: as a space maintainer, Frankel effect, headgear effect, post-maxillary expansion retention, derotation of molars, and combined using with headgear. CONCLUSION: The advantages of lip bumper are inclusions of a wide range of indication, wearing for 24 hours per day, easily fabricating and prefabricating, comfortable and hidden for patients. PMID- 12539654 TI - [Clinical application of computed arthroscope of the temporomandibular joint]. AB - OBJECTIVE: The effect of clinical application on the computed arthroscope of the temporomandibular joint (TMJ) is evaluated. METHODS: The single arthroscopic images were input into computer and memorized into magneto optical disk. With the help of Photoshop 5.0 in WINDOWS 95, the images were combined by technique of virtual process, adjusted by rotation of images, marginal blur, and chromatism correction to compound sagittal and/or coronal panoramic images of articular surfaces. According to different needs, the composite images can be printed with different printers. During the period from May 1998 to May 1999, the TMJ preoperative panoramic images were composed with computed arthroscope (CA) in the 32 joints. Of them, there were 12 joints with internal derangement (ID), 8 osteoarthrosis (OA), 8 adhesion, 2 disk perforation, and 2 synovial chondromatosis. The post-operative panoramic images were also made up to evaluate the surgical effects in 10 joints. RESULTS: In all of 32 joints, the 32 panoramic arthroscopic images of upper cavities were composed. In addition, the images of lower cavities were composed in 2 OA and 1 perforation. The postoperative panoramic images were also made up in 4 OA, 4 adhesion, and 2 synovial chondromatosis. All of the above-mentioned images showed integrally the entire structure of articular cavity, intracapsular pathologic/surgical appearances, and the relationships among the different tissues or articular surfaces. CONCLUSION: The CA can enhance the comprehensive ability of diagnostic arthroscope, and help to exchange and spread the experiences of the TMJ arthroscopic surgery. PMID- 12539655 TI - [The disinfection efficiency comparison of different treatments on dental impression and gypsum casts]. AB - OBJECTIVE: In this study, the disinfecting efficiency of five disinfecting methods to three bacterial: Staphylococcus epidermidis, Streptococcus sanguis and Bacillus subtilis were evaluated. METHODS: Germ free impressions and gypsum casts were divided into three teams contained 16 impressions and 5 gypsum casts for each one. Each team was smeared with each one of the three bacterial solutions. Then four disinfecting methods were administrated on these impressions separately, 2% glutaraldehyde immersion, 2% glutaraldehyde spray, 5% Eric immersion, 5% Eric spray. And ozone treatment was administrated on gypsum casts. Control teams were set up. After the treatment the impressions and gypsum casts were sampled at standard sites. The colonies were counted after culture and were used to deduce the germicidal ratio as the standard of disinfecting efficiency. RESULTS: There was no difference between the efficiencies of five disinfecting methods for Streptococcus sanguis and Staphylococcus epidermidis. But for B. subtilis, the immersion methods have the highest efficiency among the methods and the ozone treatment has the least efficiency. CONCLUSION: 2% glutaraldehyde immersion, spray, 5% Eric immersion, spray are all effective disinfecting methods for impressions and ozone treatment is an effective method in disinfecting the gypsum casts. PMID- 12539656 TI - [The effects of tea polyphenols on the adherence of cariogenic bacterium to the salivary acquired pellicle in vitro]. AB - OBJECTIVE: It has been proved that tea polyphenols (TP) can inhibit the sucrose dependent adherence of oral cariogenic bacterium to the tooth surface. This study is carried on to investigate the effects of tea polyphenols on the preliminary adherence of main cariogenic bacterium to salivary acquired pellicle and reveal the anticarious mechanisms of tea polyphenols further. METHODS: The hydroxyapatites were coated by saliva to form S-HA, an in-vitro model of experimental pellicle. Streptococcus mutans 3a3 and Actinomyces viscosus ATCC 19246 were chosen as the main cariogenic bacterium, and the microorganisms were radiolabelled by growing them in TPY culture medium supplemented with 3H thymidine. Tea polyphenols solution was prepared, the concentrations of which were 0.5, 1.0, 2.0 and 4.0 mg/ml respectively. Two parts of inhibited adherence tests were done with TP-treated S-HA or TP-treated bacterium. In the first one, S HA was immerged into the TP solution of different concentrations for 1 h before the attachment of bacterium. In the other, the bacterium were mixed with the TP solution for 1 h and then centrifuged and washed before bound to S-HA. The whole process was carried on in KCl buffer solution. The number of 3H-thymidine labelled bacterium which attached to S-HA were determined by scintillation counting. RESULTS: In both the two parts of this study, tea polyphenols solution, the concentrations of which were changed from 1.0 to 4.0 mg/ml, could decrease the number of S. mutans 3a3 and A. viscosus ATCC 19246, and the inhibition rates rose with the increase of the concentrations of TP solution. But the inhibitory effects of 0.5 mg/ml TP solution were inconsistent in different tests. The results showed that 1.0-4.0 mg/ml TP solution could inhibit the attachments of S. mutans and A. viscosus to S-HA effectively. The inhibitory functions of tea polyphenols were so obvious that in some experiments, while the concentrations of TP solution were changed from 2.0 to 4.0 mg/ml, the number of 3H-thymidine labelled bacterium which attached to S-HA was even less than that of the same bacterium which attached to naked HA. CONCLUSION: Tea polyphenols can inhibit the preliminary adherence of Streptococcus mutans and Actinomyces viscosus to S-HA effectively. TP may prevent enamel from caries by decreasing the adherence of main cariogenic bacterium to salivary acquired pellicle. PMID- 12539657 TI - [The effects of tea polyphenols on the adherence of cariogenic bacterium to the collagen in vitro]. AB - OBJECTIVE: In the first phase of our study, it has been proved that tea polyphenols (TP) can inhibit the preliminary adherence of oral cariogenic bacterium to salivary acquired pellicle. This study is carried on to investigate the effects of tea polyphenols on the preliminary adherence of main cariogenic bacterium to collagen I and reveal the anticarious mechanisms of tea polyphenols further. METHODS: Tea polyphenols solution was prepared firstly, the concentrations of which were 0.5, 1.0, 2.0 and 4.0 mg/ml. An in-vitro model of experimental pellicle(C-HA) was formed by immerging the hydroxyapatites in 250 micrograms/ml collagen I solution for 1 h. Streptococcus mutans 3a3, Actinomyces viscosus ATCC 19246 and Lactobacillus ATCC 4546 were chosen as the main cariogenic bacterium and radiolabelled by growing them in TPY culture medium supplemented with 3H-thymidine. Two parts of inhibited adherence tests were done with TP-treated C-HA or TP-treated bacterium respectively. In the first one, C-HA was immerged into the TP solution of different concentrations for 1 h before the attachment of bacterium. In the other, the bacterium were mixed with the TP solution for 1 h and then centrifuged and washed before bound to C-HA. KCl buffer solution provided a steady condition to the whole process. Finally, the number of 3H-thymidine-labelled bacterium which attached to C-HA were determined by scintillation counting. RESULTS: In both the two parts of this study, tea polyphenols solution, the concentrations of which were changed from 1.0 to 4.0 mg/ml could inhibit the attachments of S. mutans 3a3, A. viscosus ATCC 19246 and Lactobacillus ATCC 4546 to C-HA effectively, and the inhibition rates rose with the increase of the concentrations of TP solution. But the inhibitory effects of 0.5 mg/ml TP solution were inconsistent in different tests. The inhibitory functions of tea polyphenols were so prominent that in some experiments, while the concentrations of TP solution were higher than 2.0 mg/ml, the number of 3H thymidine-labelled bacterium which attached to C-HA was even less than that of the same bacterium which attached to naked HA. These results showed that TP had the remarkable inhibitory efficiency on the attachments of S. mutans, A. viscosus and Lactobacillus to C-HA. CONCLUSION: Tea polyphenols can inhibit the preliminary adherence of Streptococcus mutans, Actinomyces viscosus and Lactobacillus to C-HA effectively. TP may prevent dentine from caries by decreasing the adherence of main cariogenic bacterium to collagen. PMID- 12539658 TI - [The study of occlusal contact areas of upper premolars and molars in the unilateral cleft lip and palate]. AB - OBJECTIVE: Occlusal contact plays an important roll in masticatory procedure. It infers to biting force and masticatory performance. In order to analysis the oral function in UCLP, the author tested the occlusal contact areas of upper premolars and molars in UCLP. METHODS: 22 UCLP patients (male 13, female 9, permanent teeth) were selected, which had no temporomandibular joint dysfunction. All of them had received plastic surgery treatment and never received orthodontic therapy. Their occlusal contact areas of upper premolars and molars in intercaspid position were recorded by performed soft wax, and the results were managed by computer with the software of micro image analysis system. All the data were input to computer and be analyzed by statistics. RESULTS: 1. In normal people, the occlusal contact areas of upper premolars and molars had no differences in bilateral and in sexual. 2. there was no sexual difference in UCLP as that in normal group, and the occlusal contact areas in cleft side or non cleft side had no difference yet. 3. The occlusal contact areas in UCLP were lower than that in normal people. 4. The decrease of occlusal contact areas was the most important factor to low masticatory performance in UCLP patients. CONCLUSION: After plastic surgery in UCLP, orthodontic therapy will improve masticatory contact and contribute to craniofacial esthetic and masticatory function. PMID- 12539659 TI - [Determinations and analysis of masticatory performance in unilateral cleft lip and palate]. AB - OBJECTIVE: Masticatory function is one of the most important physical functions in craniomaxilla system. Masticatory performance is evaluated with the degree of broken dawn tested food in a limited period of time. Unilateral cleft lip and palate (UCLP) is a congenital abnormality in craniofacial growth and developing. It seriously damages face esthetics and masticatory function in UCLP patients. METHODS: 28 UCLP patients (male 17, female 11, permanent teeth) were selected, which all had received plastic operation treatments and never received any orthodontic therapy. The masticatory performance was determined by the advanced light absorbtion method and took peanuts as tested food. Meanwhile, malocclusal patterns and castings were identified in clinic. All of the data were treated with t-test and step wise-regression in statistics. RESULTS: 1. Masticatory performance of male was higher than that of female in normal people, but there was no sexual difference in UCLP. 2. Masticatory performance in UCLP was lower about 53.97% than that in normal people. 3. Different masticatory times within limited time had no effects on masticatory performance. 4. Different Patterns of malocclusion had different effects on masticatory performance. CONCLUSION: Orthodontic therapy is necessary to improve masticatory function in UCLP after plastic surgery. PMID- 12539660 TI - [Circadian rhythm changes of the levels of endogenous insulin in growing rat's blood after functional mandibular protrution]. AB - OBJECTIVE: In order to determine whether the metabolism of the body can be influenced by local functional orthopedic treatments and the relationship between the proliferation and translation of condyle and the metabolism of the body, the author investigates the circadian rhythm of the levels of endogenous insulin (Ins) in growing rats' blood and the changes after functional mandibular protrusion in daytime (8:00-20:00) or nighttime (20:00-8:00). METHODS: 72 4-week old SD rats were divided equally and randomly into experimental and control groups, and the rats in experimental groups were put on the functional appliances in daytime (8:00-20:00) or nighttime (20:00-8:00). After 7 days, all rats were sacrificed at 0:00, 4:00, 8:00, 12:00, 16:00, 20:00 in a day. Radioimmunoassay methods wore used to detect the levels of endogenous insulin in growing rats' blood in every four hours of a day. RESULTS: 1. The growing rats' blood displayed a significant circadian rhythm in the levels of endogenous insulin, the acrophase was about 13:02. 2. The circadian rhythm still existed after used functional appliance. No significant changes (both mesors and amplitudes) had been observed in the experimental groups comparing with the control groups. 3. The circadian rhythm of insulin in blood concerned with that of endogenous insulin in condyle. CONCLUSION: 1. The metabolism of the body can not be influenced by local functional orthopedic treatments. 2. The proliferation and translation of condyle concern with the metabolism of the body. PMID- 12539661 TI - [Properties and infiltration arts of machinable infiltration ceramic(MIC)]. AB - OBJECTIVE: The purpose of this study is to explore the infiltration arts of MIC and study the effects of different packing density of Al2O3 matrix on the properties of MIC. METHODS: alpha-Al2O3 specimens were fabricated by pouring alpha-Al2O3 slip with different powder/liquid ratios(P/L = 3.5, 7.5, 10.5) into a mold, and subsequently pre-fired at 1160 degrees C for 6 hours to form Al2O3 matrix. The packing density of the matrices were measured. Infiltration concepts were introduced into this study by infiltrating molten mica micro-crystalline glass into the porous Al2O3 matrix at 1160 degrees C for 6 hours to form a continuous interpenetrating composite. The composite then underwent micro crystallization by nucleating at 550 degrees C for 1 hour and crystallizing at 900 degrees C for 1 hour, which resulted in the MIC. Mechanical properties including three point flexural strength, elastic modulus, Vicker's hardness, indentation fracture toughness and Weibull's modulus of flexural strength were determined. Parameters of machinability(H/KIC)2 of MIC were calculated. XRD and SEM were employed to study its microstructure. RESULTS: The resulted matrices reached packing densities of 63%, 76%, 78% with P/L of 3.5, 7.5 and 10.5. The MIC attained high strength and good machinability after infiltration. Three-point flexural strength and indentation fracture toughness were 342, 431, 374 MPa and 4.05, 4.14, 5.02 MPa m1/2 for MIC with packing density of 63%, 76%, 78% separately. And parameters of machinability were 5.41, 6.84 and 7.39 respectively. Packing density of Al2O3 matrix significantly influenced the mechanical properties. Maximum properties were obtained with a matrix packing density of 75%(P/L = 7.5), with a Weibull's modulus of flexural strength of 6.8. Machinability decreased with the increase of P/L ratio. Micro-crystallizing treatment resulted in the formation of evenly distributed mica crystalline in the composite, which contributed to the high strength of this composite material. CONCLUSION: MIC is a new infiltrated ceramic with favorable strength and machinability which can satisfy the prosthodontic requirements as all ceramic crown and bridge materials, it also shows promising outlook for future developments and clinical usage. PMID- 12539662 TI - [Study on physical properties of titanium alloy sample fabricated with vacuum sintered powder metallurgy]. AB - OBJECTIVE: To investigate the physical properties of titanium alloy fabricated with vacuum-sintered powder metallurgy. METHODS: The titanium powders of three different particle sizes(-160mesh, -200 - +300mesh, -300mesh) were selected, and mixed with copper and aluminum powder in different proportions. Two other groups were made up of titanium powder(-200 - +300mesh) plated with copper and tin. The build-up and, condensation method and a double-direction press with a metal mold were used. The green compacts were sintered at 1000 degrees C for 15 minutes in a vacuum furnace at 0.025 Pa. In the double-direction press, the specimens were compacted at the pressure of 100 MPa, 200 MPa and 300 MPa respectively. Then the linear shrinkage ratio and the opening porosity of the sintered compacts were evaluated respectively. RESULTS: 1. The linear shrinkage ratio of specimens decreased with the increased compacted pressure(P < 0.05). There was no significant difference among the linear shrinkage ratios of three different titanium powders at the same compacted pressure(P > 0.05), but that of titanium powder plated with copper and tin was higher than those of other specimens without plating(P < 0.05). 2. The opening porosity of specimens decreased with the increased compacted pressure(P < 0.05). Three different sized particle of titanium powder did not affect the opening porosity at the same compacted pressure(P > 0.05). CONCLUSION: The composition of titanium-based metal powder mixtures and the compacted pressures affect the physical properties of sintered compacts. Titanium powder plated with copper and tin is compacted and sintered easily, and the physical properties of sintered compacts are greatly improved. PMID- 12539663 TI - [The quantitative analysis of cyclin D1 expression in RA-treated and non-RA treated ectomesenchymal cells in vitro]. AB - OBJECTIVE: To elucidate the alteration of Cyclin D1 protein between RA-treated and the non-RA-treated ectomesenchymal cells in embryonic mice. METHODS: Ectomesenchymal cells in the developing orofacial processes were explanted from embryonic BALB/c mice on generation day 12(15) (the 15th hour of the 12th day). Primary cells were cultured using the tissue mass method. The variants of growth curves were examined in the RA-treated and normal groups. Protein of Cyclin D1 was detected using immunohistochemistry. The positive degree of Cyclin D1 staining was analyzed using image analysis. RESULTS: The findings showed that the growth of ectomesenchymal cells was inhibited dramatically. Cyclin D1 expressed strongly in normal ectomesenchymal cells. However, its expression decreased evidently after RA-treated. There was significant difference between the test group and the normal group (P < 0.01). CONCLUSION: Low expression of Cyclin D1 may contribute to the inhibition of ectomesenchymal cells. It is necessary for Cyclin D1 to express appropriately during the normal meiosis of ectomesenchymal cells. PMID- 12539664 TI - [A study of sensory pathway in central nervous system of experimental tooth movement in rats]. AB - OBJECTIVE: To investigate whether ventral posteromedial thalamus nucleus (VPM) receive the stimulation from experimental tooth movement in rats through the conduction of spinal trigeminal caudal nucleus (Vc), thus to study the central sensory pathway of tooth movement. METHODS: 2% fluorogold (FG) was injected into rat's VPM using microinjector through retrograde tracing technique. After 4-6 days, the experimental tooth movement was conducted in contra-side against FG injection. 2 hours later, animals were sacrificed and FOS protein immunocytochemical reaction was conducted. The sections were observed under Olympus fluorescence microscope. RESULTS: FOS immunocytochemical-positive neurons were densely distributed in epilateral superfacial layer of Vc, being belt-like with more in dorsolateral 2 hours after experimental tooth movement; FG-labeled neurons were found in Vc and principle trigeminal nucleus complex, especially within the whole length of Vc, with more in layer I and the junction of layers I and II. There were both FOS immunocytochemically positive and FG-labeled neurons, approximately 5% of the whole FG-labeled neurons. CONCLUSION: As a special oro facial stimulation, there are corresponding parts in the central nervous system according to tooth movement. The stimulating message integrated in Vc and Vc has projective neurons to VPM, and VPM might be a relay station of the stimulating message of experimental tooth movement in rats. PMID- 12539665 TI - [Establishment of rat submandibular gland squamous cell carcinoma induced by DMBA]. AB - OBJECTIVE: To study carcinogenesis and development of salivary gland tumor and establish an animal model of submandibular gland (SMG) tumor. METHODS: Histopathological study during carcinogenesis in rat SMGs using (9,10-dimethyl 1,2-benzanthracene (DMBA) was evaluated. A total of 50 male and female Sprague Dawley (SD) rats of 8 weeks old and 180-200 g weight were obtained from the Animal Center of Henan Medical University. Under pentobarbital sodium anesthesia, the left SMGs were exposed by surgical procedure. A sponge pellet (1.0 mm x 1.0 mm x 1.0 mm, made by authors) was used as the carrier of the carcinogen. The sponge containing 2% DMBA (Fluka, Switzerland)/acetone solution was implanted into the glandular tissue of the left SMGs. Four rats (2 males and 2 females) were killed after every 2 weeks of the DMBA/sponge implantation. The same method of sponge implantation without DMBA was used at the right side of SMG as a control. All rats left were killed after 20 weeks. The SMGs were fixed in 10% formalin buffer solution for 24 hours, and embedded in paraffin, then 4 microns thick sections were made for histopathological study. RESULTS: The earliest tumor occurred after 4 weeks of implantation of sponge, a total of 21 lateral tumors were induced (10 females, 11 males). There was no tumor found in the controlled SMG. The peak time of tumor genesis was after 8-12 weeks of implantation, all tumors induced were squamous cell carcinomas(SCCs). The induced tumors grew slowly below the mandibles of rats, which were present as nodular masses without capsules, and the borders were not clear. They were slightly hard when palpated. The process of carcinogenesis can be described as following: squamous metaplasia of cyst-like structures occurred, then SCCs were induced and invaded surrounding tissues. No metastasis was observed in regional lymph nodes and other organs. CONCLUSION: SCCs of SMG can be induced by implantation of DMBA. The present study supports the conclusion that all duct segments undergo squamous metaplasia, and therefore may participate in the genesis of neoplasia during experimental carcinogenesis. PMID- 12539666 TI - [Optimal treatment parameters for damage of chicken comb vessels using chlorophyll derivative IV in photodynamic therapy]. AB - OBJECTIVE: To define the optimal treatment parameters of chlorophyll derivative IV (CPD4) in photodynamic therapy(PDT) of port wine stains(PWS) by observing the morphological changes of chicken combs. METHODS: 102 animals were divided into 6 groups at random, including 3 control groups(given no treatment, given light without photosensitizer or given photosensitizer without light) and 3 experimental groups treated with different CPD4 dosages(5, 7.5, 10, 12.5, 15 mg/kg body weight), power density(50, 100, 150, 200 mW/cm2), energy density (15, 30, 45, 60, 120, 135, 180 J/cm2) and different intervals between CPD4 injection and irradiation (immediately, 5, 10, 20, 30, 60, 120, 180, 240 minutes). The main emitting peak of light source was at 650 nm, and CPD4 was injected intravenously, then the appearance of chicken comb was recorded everyday. The prepared sections were stained with HE. The histological changes of chicken comb were observed by light microscope. RESULTS: The control animals showed no significant comb changes, while the PDT could cause blanching of the chicken comb in the experimental group. The histologic changes included damage of erythrocytes and endothelial cells, vascular rupture, hemorrhage or disappearance, denaturation of perivascular collagen fiber and local necrosis of chicken combs. The treatment parameters which caused the most obvious blanching of the chicken comb were 7.5 10 mg/kg body weight of CPD4 dosage, 100-150 mW/cm2 of power density, 60-90 J/cm2 of energy density, and the intervals between CPD4 injection and irradiation were 0-20 minutes. CONCLUSION: CPD4 in PDT can cause selective damage of dermal capillaries in chicken comb and blanching of the chicken comb with optimal treatment parameters. The results will offer the basis for further investigation of clinic treatment parameters. PMID- 12539667 TI - [Effect of calcium on adherence of Streptococcus mutans MT6R(serotype c) surface protein P1]. AB - OBJECTIVE: To study effects of calcium on the adherence of Streptococcus mutans MT6R(serotype c) surface protein P1 to saliva-coated hydroxyapatite(S-HA). METHODS: The surface protein P1 of Streptococcus mutans MT6R was purified by PAGE and labeled with 131I(131I-P1). The adherence amount of 131I-P1 to S-HA in different calcium concentration liquid was measured. RESULTS: The amount of 131I P1 adherence to S-HA in 0, 0.1, 0.25, 0.5, 1.0, 1.5, 2.0 mmol/L, and calcium was 7175 + 183, 7516 + 192, 8850 + 215, 9335 + 204, 10087 + 228, 10179 + 224 and 10200 + 317 cpm, respectively(P < 0.01). The calcium concentration from 0 to 1.0 mmol/L, the amount of 131I-P1 adherence was increased statistically(P < 0.05), calcium concentration from 1.0 mmol/L to 2.0 mmol/L, the amount of 131I-P1 adherence was increased unstatistically(P > 0.05). CONCLUSION: Calcium promoted the adherence of surface protein P1 to the S-HA. This result suggested that calcium was involved in the adherence of Streptococcus mutans MT6R (serotype c) surface protein P1. PMID- 12539668 TI - [Classification of eyelid and designs of the double-eyelid operation]. AB - OBJECTIVE: To discuss the morphological characters of eyelids and the designs of double-eyelid operation on various eyelids. METHODS: The classification of eyelids was named according to morphological characters of eyelids from 6847 cases. A survey of 92 cases treated by double-eyelid operation was carried out to compare the external eye morphology of pre-operation with that of post-operation. RESULTS: There were several types of eyelids, which were morphologically classified according to eyes' length and height, eyelid skin thickness, eyeball protrusion and eye shape, including eyelids with long and narrow eyes, eyelids with thin and soft skin, eyelids with thick subcutaneous tissue, eyelids with small and round eyes, eyelids with protruding eyeball, and eyelids with triangle eyes. The aesthetic effects of double-eyelid operation on all these types of eyelids were evaluated. CONCLUSION: The indication of double-eyelid operation should be considered correctly, and the design should be made according to classification of the morphological characters of eyelids. Double-eyelid operation may be not appropriate for the eyelids with small, round and protruding eyes. PMID- 12539669 TI - [Simultaneous intrusion and retraction of the anterior teeth using a three-piece base arch]. AB - OBJECTIVE: To evaluate the effects of the three-piece base arch on overbite correction of Class II malocclusion. METHODS: 20 patients with high angle, flared incisors were treated using a three-piece base arch appliance. The intrusion force of four upper incisors was adjusted to approximately 50 g. The line of force action was 2 mm distally to the resistant center(RC) and the retraction force was 20 g, the right and left posterior segments were joined by a palatal bar. Cephalograms were taken before treatment (T1) and six months after treatment (T2). RESULTS: The upper molars moved mesially 0.60 +/- 0.35 mm and the distance of the vertical extrusion was 0.80 +/- 0.52 mm. The distances of the upper central incisor retraction and intrusion were -4.20 +/- 2.12 mm and 3.10 +/- 0.54 mm respectively. The RC of the central incisor retracted -4.12 +/- 1.96 mm and intruded 3.20 +/- 0.66 mm. The axial inclination of the upper incisor-palatal plane changed from 123.21 degrees +/- 4.26 degrees to 116.00 degrees +/- 3.96 degrees. CONCLUSION: The three-piece segmented approach can effectively intrude and retract the upper anterior teeth for flared incisors and deep overbite. PMID- 12539670 TI - [Clinical study of direct digital radiography in caries detection]. AB - OBJECTIVE: Direct digital radiography is a computer-based image technique, the purpose of this paper is to investigate possible usage of direct digital radiography-radio visiography (RVG) in caries detection in clinic. METHODS: A human head model with teeth was used to establish the operative exposure time for RVG in vitro, then RVG images for normal and carious tooth were obtained in vivo and were compared with conventional E-speed film in image quality and diagnostic accuracy. RESULTS: The optimal exposure time for RVG image was obtained, and the exposure time was different in each tooth quarter. The acceptable rate of image quality and diagnostic accuracy of RVG was statistically equivalent to that of E speed film (P > 0.05), however, the excellent rate of RVG was statistically superior to that of E-speed film (P < 0.05). CONCLUSION: The conventional X-ray will be substituted by RVG in caries detection in clinic. PMID- 12539671 TI - [The investigation of color selection of 4340 cases of ceramic restorations]. AB - OBJECTIVE: To find color coordinates of Vita color tabs compare with Chinese natural teeth and summarize the skills of color matching. METHODS: We studied 15,836 ceramic restorations of 4340 cases from 138 clinics in 18 provinces throughout China and compared their shade with that of the adjacent natural teeth using the Vita shade guide which has 16 standard tooth colors. Statistical analyses was made by comparing aptness of Chinese teeth with Vita shade. RESULTS: We found that 64.47% of the restorations were matched by Vita A, 15.85% were matched by Vita B, and 19.68% were matched by Vita C and D. The arrangement of Vita 16 colors by using frequency was A2, A3, B2, A1, A3.5, C1, D2, D3, C2, B3, C3, B1, A4, D4, B4 and C4. The first five color matching tabs used most frequently covered 73.85% of the total, while the last four colors only covered 3.07%. It seemed that Vita color tabs could not match well with Chinese teeth. We found that the shade of natural teeth was closely related to sex. But there were no significant difference between tooth color and different areas. CONCLUSION: Tooth color is closely related to sex. The color of ceramic crowns in men are darker than that in women. We have evidences that there are clear differences in color distribution between Chinese natural teeth and Vita shade guide. PMID- 12539672 TI - [The effects of CDFI and CDE high-frequency ultrasonography on diagnosis of parotid neoplasm]. AB - OBJECTIVE: To evaluate the effects of high-frequency color ultrasonography on diagnosis of parotid tumors. METHODS: 36 cases of parotid benign or malignant tumors were observed from margin to inner with color Doppler flow imaging (CDFI) and color Doppler energy (CDE) according to tumors' two dimention. RESULTS: The features of benign tumors included round or round-like shapes, clear margins and inner balanced echogram. The margin feature of CDFI was star-like images, and the peak systolic celocity was up to 18.6 and 16.2 cm/s. The coincidence of diagnosis was 90%. Meanwhile, the shapes of malignant tumors showed irregular and incomplete mass and asymmetrical inner echogram. The primary characteristics of CDFI were branch-like or basket-like images, and the peak systolic velocity was much higher (35.2 and 32.4 cm/s) than that of benign tumors. There was significant difference between these two groups (P < 0.05). It indicated richer flow signals in CDE than in CDFI. CONCLUSION: CDFI and CDE show an important clinical value in diagnosing parotid neoplasm. PMID- 12539673 TI - [Improvement on the individual operative design of unilateral cleft lip repair]. AB - OBJECTIVE: To study the difference of operative designs in unilateral cleft lip repair according to characteristic deformity of each patient with unilateral cleft lip. METHODS: The distance from both alar bases and midpoint of columella base to both peaks of Cupid's bow were consulted is equal as standard design model. The mark point and incision of individual unilateral cleft lip repair were designed using geometrical analysis. RESULTS: The new operative design technique included that the end point of rotational incision should be inside of the cross point from the line dividing both Cupid's bow peaks and philtrum on the non-cleft side, and the other design was that the end point of alar base on the cleft side located at the cross point from the horizontal line that passed the midpoint of alar base on the non-cleft side to meet the alar base on the cleft side. Depending on the above thesis, the new operative methods were created for unilateral cleft lip repair. One of the new individual operative designs was similar with Millard's method in the shape of operative design, but there was a big difference on the principle of marking the columella base and alar base on the cleft side between individual operative designs and Millard's design. This new operative design only suit for unilateral cleft lip with symmetric distances from alar base to Cupid's bow peaks in both sides. Another individual operative design added a new incision on the cleft side comparing with the former individual operative design, which could be used in unilateral cleft lip when the distance from alar base to Cupid's bow peak on the cleft side was shorter than that on the non-cleft side. CONCLUSION: The new individual operative designs of unilateral cleft lip repair have been created, and the important advantages of these new individual operative designs are that each marking point can been decided clearly according to special mathematical principles. Each operative design only suit for one type of unilateral cleft deformity, so the types of deformity in unilateral cleft lip can been also classified depending on the different distances from alar base to Cupid's bow peak between both sides. This classification of unilateral cleft lip only depends on operative designs, and these new individual operative designs of unilateral cleft lip repair are easy to be understanded and applied, as well as easy to obtain stable result after operation. PMID- 12539674 TI - [Cephalometrical analysis of effects of premaxillary orthopedic treatment on maxillary growth of Wistar rats with bilateral artificial cleft palate]. AB - OBJECTIVE: To measure the differences between the experimental group and the two controlled groups by using the cephalometric radiography technique to study the effects of premaxillary orthopedic treatments on the growth and development of maxilla. METHODS: To make an animal model with bilateral cleft palate by surgery operation: General anesthesia was carried on by injecting 2.5% pentobarbital (25 mg/kg) into abdominal cavity, then almost the same bone of rats' palate were removed from bilateral palate. An appliance, based on the principle of Latham's appliance was made and designed for premaxillary orthopedic treatment. It was fixed in rats' palate using pins under the condition of general anesthesia. After 20 days of orthopedic treatment, the rats were anesthetized by ether, and cephalometric films were taken under self-designed cephalometer and analysed aided by computer. RESULTS: The distances that represented the maxillary position in the posterior-anterior (PA) direction significantly increased (P < 0.05), while the angles that represented the direction of premaxillary growth significantly decreased (P < 0.05) between the experimental group and the two controlled groups. There were no significant difference between the two controlled groups. CONCLUSION: Premaxilla of rats can be effectively moved backward and rotated downward and backward on the PA direction by appliance. The appliance can move the segment palate forward during the premaxillary backword. The appliance can stimulate maxillary growth. PMID- 12539675 TI - [An cephalomectric study on craniofacial structures of Uygur adults with normal occlusion in Xinjiang autonomous region]. AB - OBJECTIVE: To study craniofacial structures of Uygur adults with normal occlusion and establish the standard cephalometric values to evaluate the craniofacial soft and hard tissue structures of normal Uygur adults. METHODS: Lateral cephalometric radiographs of 93 Uygur adults with normal occlusion (43 men and 50 women) were evaluated with the same standard. Their age ranged from 18 to 25 years old, and they had normal and well-balanced faces without receiving any orthodontical treatment. The lateral cephalogram of each normal adult was traced digitized and measured by the same computer system, then the indexes of craniofacial structures were analyzed by multifactor cluster analysis. 18 typical items were chosen to analyze the characters of the craniofacial structures of Uygur adults with normal occlusion. At the same time, Bayers discriminant analysis was used to identify and evaluate the cluster results. RESULTS: Uygur adults with normal occlusion were divided into three patterns according to the multifactor cluster analytic results: horizontal tendency pattern (40%), medial pattern (43%) and vertical tendency pattern (17%). When the horizontal tendency pattern was compared with the medial pattern, it showed that the height of S-Go increased (P < 0.05), while the height of ANS-Xi-Pm decreased (P < 0.05); The proportion of S-Go/N-Me increased (P < 0.01). The angle of Ar-Go-Me and Y axis decreased (P < 0.05), which meant that the mandible rotated in the counter-clockwise direction. SN-MP, PP-MP, OP-MP, MP-FH all decreased (P < 0.01), which meant that the SN plane, the PP plane, the occlusion plane and the MP plane all had the trend of hypodivergence. When the vertical tendency pattern was compared with the medial pattern, the height of Ar-Fo decreased (P < 0.01), while the height of ANS-Me increased (P < 0.01); The angle of Ar-Go-Me, Y axis, SN-MP, PP-MP, OP-MP and MP FH all increased (P < 0.01). These results showed that the mandible rotated in the clockwise direction and the SN plane, the PP plane, the occlusion plane, and the MP plane all had the trend of hyperdivergence. At last, according to the different skeletal patterns, the normal cephalometric values of Uygur adults with normal occlusion were established. CONCLUSION: Significantly craniofacial structure difference exist in the adults with normal occlusion. Therefore, it is necessary to establish the normal cephalometric values of Uygur adults with normal occlusion according to different skeletal patterns. PMID- 12539676 TI - [In vitro studies about the effects of implanted materials on osteocalcin and alkaline phosphatase of rat osteoblasts]. AB - OBJECTIVE: To evaluate the effects of the four implant materials, hydroxyl poly calcium sodium phosphate (HPA), hydroxyapatite (HA), bioglass ceramics (BGC) and titanium (Ti) on osteocalcin secreting amount and alkaline phosphatase activity of rat osteoblasts cultured in vitro, and expose the biological compatibility conditions of the implant materials as well as the index to estimate cytocompatibility. METHODS: The powders of HPA, HA, BGC and Ti were sterilized and made into extracted solutions with 199 culture medium, and the isolated osteoblasts which were taken from three-day-old Sprague-Dawley rats' calvaria, were incubated in 199 culture medium too. Then the osteoblasts were mixed and incubated into the four kinds of extracted solutions of the implant materials. After eight days of co-incubation, the cells were taken out to measure alkaline phosphatase activity with velocity analysis, and the extracellular liquid was used to estimate osteocalcin secreting amount with radioisotope analysis. RESULTS: 1. Osteocalcin secreting amount of the osteoblasts co-incubated with HPA, HA, BGC were as normal as the control, while that of other cells co incubated with titanium was reduced (P < 0.05). 2. Alkaline phosphatase activities of all the co-incubated osteoblasts were as normal as the control. CONCLUSION: The osteocalcin secreting amount and alkaline phosphatase activity can reflect the biological compatibility of implant materials, and they may be used as indexes to evaluate biological compatibility condition of implant materials. PMID- 12539677 TI - [Survey of the patients with oblique facial clefts in China]. AB - OBJECTIVE: A descriptive study was conducted to investigate the epidemiological characteristics of oblique facial clefts in China. METHODS: From 1987 to 1992, data were collected from more than 500 hospital-based units involved in a monitoring program and were analyzed by the National Center for Birth Defects Monitoring. The monitored subjects were the hospitalized births from 28 weeks gestation to a period within 7 days after delivery. RESULTS: There were 71 cases of oblique facial clefts in 4,489,692 births with an incidence of 0.16/10(4) in China. The incidence of oblique facial clefts in the rural and urban areas were 0.19/10(4) and 0.14/10(4) respectively, 0.18/10(4) in males and 0.12/10(4) in females. No significant differences were found. The unilateral oblique facial clefts were counted for 80%, and bilateral cases 20%. The incidence of unilateral oblique facial clefts in males is higher than that in females. The perinatal mortality was as high as 51%. There are 76% cases with associated abnormalities, most of which involved the maxillofacial region. The most common associated abnormality is cleft lip with cleft palate. CONCLUSION: The incidence of oblique facial clefts in China is the same as those in other countries. There is an unilateral predominance in this defect, especially in males. The perinatal mortality of this defect is high. PMID- 12539678 TI - [Preparation of cucurbitacinBE polylactic acid nano-particles for targeting cervical lymph nodes]. AB - OBJECTIVE: Anti-cancer nano-particles for targeting cervical lymph nodes were designed and prepared on the basis of the physiologic and anatomic properties of the peri-oral cancer lymphatic capillaries. METHODS: The biodegradable polylactic acid (PLA) was selected as the carrier material, and cucurbitacinBE (CuBE) with very high anticancer activity against oral squamous cell carcinoma was used as the model drug. According to the criteria of the diameter of the nano-particles, drug loading (DL) and embedding ratio (ER), the preparing technology was optimized by uniformly designing experiment on the basis of single-factor experiment. CucurbitacinBE poly-lactic acid nano-particles (CuBE-PLA-NP) was prepared with the method of emulsion solvent evaporation, and then CuBE-PLA-NP lyophilization injection was prepared by using 10% manninositose as the supporting agent. RESULTS: The diameter distribution for optimized CuBE-PLA-NP lyophilization injection ranged from 47 nm to 120 nm, and the average diameter was 85 nm. DL and ER were 23.03% +/- 0.47% and 93.00% +/- 0.10% respectively. The release study of the thermostatics also demonstrated that Higuchi equation was adequate to fit the release curve of CuBE-PLA-NP. CONCLUSION: CuBE-PLA-NP can be used as the release agent, as CuBE-PLA-NP can enter peri-oral cancer lymphatic capillaries for targeting cervical lymph nodes by the peri-cancer submucosal injection. Therefore, it can also used for animal and clinical experiments. PMID- 12539679 TI - [A study on mRNA expression levels of integrin beta 1 during the healing process of mandibular fractures]. AB - OBJECTIVE: The aim of this study was to investigate the spatial and temporal mRNA expression changes of integrin beta 1 during the healing process of mandibular fractures. METHODS: The in situ hybridization was performed using digoxigenin labeled integrin beta 1 mRNA oligonucleotide probe. During the healing process of mandibular fractures, the staining pattern of beta 1 mRNA was examined both spatially and time-dependently. RESULTS: Integrin beta 1 was widely present in the cells of the fractured bone, especially in osteoblasts. The positive staining was observed in the cytoplasm and nucleolus of osteoblasts. After 7 days of fracture, the expression level of integrin beta 1 increased. The peak appeared between the 14th and the 30th day, and almost recovered to normal between the 60th and 90th day. CONCLUSION: During the healing process of mandibular fracture, the expression level of integrin beta 1 in bone, especially in osteoblasts rose. It is suggested that integrin beta 1 may play an important role in the fracture healing process. PMID- 12539680 TI - [Expression changes of nerve growth factors (NGF) in pulps of injury teeth]. AB - OBJECTIVE: The aim of this study is to investigate the expression and functions of nerve growth factors during teeth injury. METHODS: Twenty male Wistar rats were given mechanical irritation in upper first molar and their molecular and biological changes were examined in different periods. Four of them were killed immediately after the teeth were irritated, the other four were killed 3 days, 5 days, and 9 days respectively after the teeth were stimulated. The perfusion was performed through the left ventricle with 4% paraformaldehyde and 0.2% picric acid in 0.1% phosphate buffer. The maxilla was decalcified and embedded. Sections of 15 microns in thickness were cut with freezing macrodome in the sagittal planes. Then the sections were stained with immunohistochemical method to observe changes of NGF in the injured dental pulp, comparing with that in the normal dental pulp. In the end, we observed the NGF distribution in the pulp under optical microscope, and analyzed the expression level of NGF with the image pattern analysis instrument. The results were statistically analyzed. RESULTS: In the normal group, the NGF expression was not observed. In the experimental group of zero hour, the NGF expression could be observed, but was very low. In the experimental group of three days, the NGF expression increased greatly, and the positive sites mainly distributed in the odontoblastic and the sub-odontoblastic layer. They also distributed in dentinal tubules. But in the group of 5 days, the level of NGF decreased. In the group of 9 days, it was the same as that of the normal group. Using the image pattern analysis instruments and statistical analysis, the results clearly showed that the expression level were lower in the group of three days than that of the other groups, and the expression levels of the groups 0 and 5 days were lower than that of the normal control group, however, that of 9-day group was the same as the normal group. CONCLUSION: NGF may play a role in the reaction and nerve restoration after teeth injury and, the expression change in different stages. PMID- 12539681 TI - [The influences of crystallized compositions in the porcelain on bonding strength of titanium to porcelain]. AB - OBJECTIVE: Sufficient porcelain-titanium bond is a vital factor determining the clinical performance of titanium-porcelain restorations. The purpose of this study was to investigate the effects of self-preparation La-porcelain composition on the porcelain-titanium bonding strength and to compare with the Vita Titankeramik. METHODS: The present study examines 5 different recipes of porcelain by weight%: SiO2, 12%-17%; LaO2, 7%-10%; Al2O3, 9%-14%; B2O3, 23%-31%; CaO, 6%-8%; K2O, 2%-3%; SrO, 2%-4%; Na2O, 1%-3%; SnO2, 8%-10%; ZrO2, 3%-5%; TiO2, 6%-8%. Specimens were tested in push type shear with a universal testing machine. Scanning electron microscopy (SEM) and electron probe microanalyzer (EPMA) were employed to reveal the microstructures and diffusion of elements in the interfacial regions between the porcelain coating and titanium to the bond strength when fired at 800 degrees C. RESULTS: The ratios of crystallized compositions had significant influences on the porcelain-titanium bond strength (P < 0.05). La-porcelain had the highest shear bond strength (37.76 MPa). The shear bond strength of the Vita Titankeramik to titanium was 20.18 MPa. The results of SEM revealed integrity of porcelain-titanium joints in La-porcelain and a greater amount of porosity in the interface of Vita Titankeramik to titanium. EPMA analysis demonstrated the aggregation of Si and Sn in the interfacial regions and their diffusion into the titanium. CONCLUSION: Chemical compositions of porcelain and ratios of crystallized compositions play the important role in the titanium porcelain bond. La-porcelain had the highest shear bond strength and good porcelain-titanium joints. La-porcelain is a new-style low fusing porcelain/titanium system. PMID- 12539682 TI - [Expression of bcl-2 and Bax genes in facial neurons and their regulation of neuron apoptosis following facial neurotmesis in rats]. AB - OBJECTIVE: The aim of this study is to investigate the relationship between the expression changes of bcl-2 and Bax genes and their functions in facial neuronal apoptosis after amputating the facial nerve. METHODS: Rats were randomized into two groups, including the experimental and the control group. The changes of expression of bcl-2 and Bax genes in the facial neurons and facial neuronal apoptosis were observed after the facial nerve amputation using the methods of ABC immunohistochemical staining and TUNEL. RESULTS: The expression of bcl-2 and Bax genes and the apoptosis of facial neurons were significantly higher in the experimental group than those of the control group from the 3rd to 21st day after amputation, but the bcl-2/Bax significantly decreased compared with that of the control group. CONCLUSION: This indicated that bcl-2 and Bax might play an important regulation role in the process of facial neurons apoptosis following facial nerve amputation. PMID- 12539683 TI - [A comparative morphometrical study on development of anterior septonasal accessory organs in the cleft palate mice]. AB - OBJECTIVE: The aim of this study was to investigate whether cleft palate is associated with the mal-development of the vomer, Jacobson's organ and para septal cartilage. METHODS: Sixty craniomaxillary complexes, which came from 30 pairs of trisomy 18 with cleft palate (Ts + CP) and euploid mouse fetuses were precisely orientated in the coronal plane, and serially sectioned at 7 microns thickness. With the aid of computer imaging analysis system the vomer in coronal and sagittal direction, Jacobson's organ and para-septal cartilage in the boundary plane of the primary and secondary palate were measured and compared quantitatively. RESULTS: The hard tissues (including the vomer and para-septal cartilage) of the anterior nasal septum in Ts + CP foetuses show serious developmental hypoplasia, however, the Jacobson's organ had no significant different between the cleft and non-cleft samples. CONCLUSION: Development hypoplasia of these hard tissues may be more closely related with the autosomal trisomic condition more than the cleft palate. PMID- 12539684 TI - [A study of millimeter wave's clinical and immunological effects on oral lichen planus patients]. AB - OBJECTIVE: The aim of this study is to observe clinical effects of the millimeter wave on oral lichen planus (OLP) and its immune mechanisms. METHODS: 30 patients with OLP were randomized into the millimeter wave group and the control group. 15 patients in the millimeter wave group were treated with millimeter wave (7.1 mm, 43 GHz, 10 mW/cm2) radiation, while the other 15 patients in the control group were just given soothing treatment. Either before or after the therapy procedure, the clinical manifestations and the T-cell sub-grouping in the peripheral circulation of each patient were double-blindly evaluated. RESULTS: The pain soothing effect in the patients treated with millimeter wave was much more obvious than that of the control group (P < 0.01). The peripheral blood CD8+ percent of millimeter wave group decreased greatly (P < 0.05), while no obvious CD8+ change was observed in the control group (P > 0.05). The millimeter wave group's CD4+/CD8+ improvement was also much more obvious than that of the control group (P < 0.05), and its CD4+/CD8+ ratio was completely recovered to the normal level. CONCLUSION: Millimeter wave can effectively relieve OLP patients' pain, and can regulate OLP patients' cellular immune condition. Millimeter wave might provide a new treatment method for OLP. PMID- 12539685 TI - [Relationship between Streptococcus mutans, Lactobacillus spp. and lactate producing level and nursing bottle caries]. AB - OBJECTIVE: The study aimed to investigate the levels of Streptococcus mutans and Lactobacillus spp. as well as the relationship between lactate-productive and aciduric organisms and nursing bottle caries. METHODS: Totally, 30 children of 2- to 2.5-year-old were divided into two groups, including the group of nursing bottle caries and the group of caries-free. Streptococcus mutans and Lactobacillus spp. were isolated from dental plaque of all the children. The amount of lactate produced was measured with a microlitre plate reader. RESULTS: The isolation frequency of Streptococcus mutans and Lactobacillus spp. were both 100 percent in the children with nursing bottle caries, and that the composition of these bacteria from nursing bottle caries lesions was higher than that of caries-free dental surface. Lactate-producing level was higher in the caries group than that of the caries-free group. CONCLUSION: This study suggests Streptococcus mutans and Lactobacillus spp. may be the major pathogenic bacteria leading to nursing bottle caries. PMID- 12539686 TI - [Clinical application of three-dimensional spiral CT in the complex maxillofacial fractures]. AB - OBJECTIVE: The aim of this study was to evaluate the clinical application of the three-dimensional spiral CT in the complex maxillofacial fractures. METHODS: A total of 26 patients with complex maxillofacial fractures were examined using spiral CT with the slice thickness of 3 to 5 mm. The surface shaded display (SSD) method was used for reconstruction of the three-dimensional (3D) images. The three-dimensional images of maxillofacial fractures were obtained by using various rotations. Before the three-dimensional images were reconstructed, the drawing and deleting way was used for these mandibular fractures. RESULTS: The method demonstrated clearly the three-dimensional anatomy and their relationships of the fractures; using the drawing and deleting way, location and move condition of mandibular fractures, especially condylar fractures, were showed clearly. CONCLUSION: The three-dimensional CT is important in evaluating complex maxillofacial fractures. It could be used to demonstrate clearly locations and move conditions of complex maxillofacial fractures and operative plans. PMID- 12539687 TI - [A study of computer-aided drawing system for individualized ideal arches]. AB - OBJECTIVE: The aim of this study is to devise a computer aided drawing system for individualized ideal arches, to resolve the discommode of manual drawing of Bonwill-Hawley arch charts and and to avoid the non-matching problem when using the universal arch charts. METHODS: On the basis of drawing principle of Bonwill Hawley individualized ideal arches, the computer program was written with Visual Basic version 6.0. RESULTS: The new computer-aided drawing system of individualized ideal arches was based on Windows9X/NT. After the data of mesiodistal diameters of maxillary anterior teeth crown (from mesial surface to distal surface of bracket for canine) were input, a matching individualized ideal arch chart would be drawing using the designed program immediately. CONCLUSION: The computer-aided drawing system of individualized ideal arch can help to draw a matching individualized ideal arch accurately and quickly. This system eliminates the complex and cockamamie process of manual drawing. PMID- 12539688 TI - [Expression of transforming growth factor beta(TGF-beta) subtypes in oral squamous cell carcinoma]. AB - OBJECTIVE: The objective of this study was to determine the expression of transforming growth factor beta (TGF-beta) subtypes and their relationship with the mechanisms of squamous cell carcinoma (OSCC) growth. METHODS: Totally 40 cases of surgical specimens of OSCC resected between 1998 and 2000 and 20 cases of normal human oral mucosa were investigated. Strepto-adridinibiotin complex (SABC) immunohistochemical staining was used to analyze the expression of TGF beta protein subtypes and their relations with clinical prognosis of OSCC. RESULTS: Semi-quantitative analysis revealed that the subtypes 1, 2 and 3 of TGF beta protein could be found in OSCC cells and normal oral epithelial cells, however the intensity of protein expression was different. Comparing with those in normal oral mucosa epithelial cells, the subtypes 1 and 2 of TGF-beta were over-expressed in OSCC cells. The over-expression of subtypes 1 and 2 of TGF-beta protein were associated with their pathological grades, clinical stages and neck lymph node metastasis (P < 0.05), whilst the subtype 3 protein of TGF-beta was not. CONCLUSION: It will be useful to detect the expression of TGF-beta subtypes in OSCC, as the subtypes 1 and 2 of TGF-beta may play an important role in OSCC growth and metastasis. PMID- 12539689 TI - [Acute toxicity and local stimulate test of cucurbitacinBE polylactic acid nano particles of targeting cervical lymph nodes]. AB - OBJECTIVE: The aim of this experiment was to study acute toxicity and local stimulation of cucurbitacinBE poly-lactic acid nano-particles (CuBE-PLA-NP) for targeting cervical lymph node. METHODS: Various doses of CuBE-PLA-NP lyophilization injection and cucurbitacinBE (CuBE) were hypodermically or veno injected into the mice, and then the drug toxicity and side effects were observed; 50% lethal dose (LD50) was counted. CuBE-PLA-NP lyophilization injection and CuBE were also injected into the quadriceps femoralis muscle of rabbits to observe the local stimulate responses of drugs. RESULTS: The fifty percent lethal doses of subcutaneous and venous CuBE-PLA-NP in mice were 16.74 mg/kg +/- 1.83 mg/kg and 5.60 mg/kg +/- 0.83 mg/kg respectively, that were higher than those of CuBE which were 8.63 mg/kg +/- 1.00 mg/kg and 2.50 mg/kg +/- 0.30 mg/kg respectively. Local stimulate response in rabbit quadriceps femoralis muscle after CuBE-PLA-NP and CuBE injected are 0 grade and 0.1 grade respectively. CONCLUSION: The acute toxicity of CuBE-PLA-NP is lower than that of CuBE. Both CuBE-PLA-NP and CuBE are adequate for local injection, but the local stimulation of CuBE-PLA-NP is smaller than that of CuBE. PMID- 12539690 TI - [Effects of XW630 on mechanical properties and trabecular structure parameters of bone tissue in ovariectomized rats]. AB - OBJECTIVE: The study aimed at investigating the effects of a new anti osteoporotic drug, XW630 on promoting the osteogenic activity of bone tissue in ovariectomized rats. METHODS: Thirty-six female SD rats, three months old, were randomly divided into four groups: sham-operated group (sham), the ovariectomized group (OVX), the esterone-therapy group (OVX + CFT) and the XW630-therapy group (OVX + XW630). Three rats in each group were killed on the 30th, 60th, and 90th day after the operation. The femur of one side was taken for the three-point bending resistance test, and the bilateral tibias were taken for the HE stain and scanning electronic microscopy (SEM) examination. RESULTS: The HE and SEM results indicated that the bone loss appeared in the OVX group 30 days after the operation, which lasted for the whole observing period. While after application of ester-one and XW630, there were different degrees of bone repairing around the trabeculae, and the osteogenic activity was obviously active in the XW630 group than that in the esterone group. During the experimental period, the three-point bending resistance of the OVX group gradually decreased and there was a significant difference compared with that of the sham group (P < 0.01), while the three-point bending resistance of the two therapy group gradually increased and there was significant difference between the OVX group and the XW630 group (P < 0.05 and P < 0.01). CONCLUSION: XW630 can effectively promote osteogenic action and prevent osteoporostic fracture in ovariectomized rats. PMID- 12539691 TI - [Regulation of XW630 on TGF-beta 1 and collagen I gene expression in osteoblasts]. AB - OBJECTIVE: The purpose of this part of study was to investigate the mechanism of a new anti-osteoporotic drug, XW630 on promoting osteogenic activity. METHODS: Total RNA were isolated from osteoblasts of the mice after the reaction with XW630 in 24 and 72 hours, that were hybridized with the extracted and digoxigenin labeled TGF-beta 1 and the collagen I cDNA probes. RESULTS: XW630 significantly promoted TGF-beta 1 and collagen I gene expression in osteoblasts, and this drug was superior than other two kinds of estrogen in promoting osteogenic activity (P < 0.05). CONCLUSION: XW630 promote osteogenic activity of osteoblasts, probably by enhancing the mRNA expression of TGF-beta 1 and collagen I. PMID- 12539692 TI - [Expression of bone morphogenetic protein (BMP)-2, 4 in mouse embryonic tooth during the bud stage]. AB - OBJECTIVE: The purpose of this paper was to investigate the distributions of bone morphogenic protein (BMP)-2, 4 in the developing molars of mice during the bud stage and, the functional relationship between BMP-2 and BMP-4. METHODS: The distribution of BMP-2, 4 were examined in developing mouse molars on the 14th (the 14th embryonic day), using monoclonal antibodies and immunohistochemistry methods. RESULTS: BMP-2, 4 were detected both in the epithelial cells and dental mesenchyme cells. CONCLUSION: There is a similar distribution pattern between BMP 2 and BMP-4. Therefore, there may be a functional relationship between BMP-2 and BMP-4 during the development of mouse embryonic teeth in the bud stage. PMID- 12539693 TI - [A computer-aid study on the craniofacial features of Archang race in Yunnan province of China]. AB - OBJECTIVE: Archang race is one of special minoritis in Yunnan province of China. However, there is not former report about overall craniofacial features in Archang race, therefore a computerized measurement on craniofacial features of Archang race was established in this study. METHODS: On the basis of the principles of random sampling, the craniofacial features of 196 normal adults of Archang race coming from Longchuan County, Dehong Dai Jingpo nationality autonomous prefecture in Yunnan province of China were investigated using the craniofacial video-computerized measurement system which was developed by us. RESULTS: Totally 41 measured items and 17 indexes of craniofacial features were obtained from male and female adults of Archang race. On the basis of 5 indexes, the people were classified according to the head, facial, and nose forms. CONCLUSION: The results of this study are of great importance in terms of that the reference is not only significant for anthropology, ethnology, but also for medicine and industry. PMID- 12539694 TI - [Cell proliferation and programmed cell death in tooth development]. AB - OBJECTIVE: To evaluate the role of cell proliferation and programmed cell death (PCD) in the tooth development by detecting programmed cell death and expression of proliferating cell nuclear antigen (PCNA). METHODS: Expression of PCNA and PCD were detected by immunohistochemical staining and Tdt-mediated dUTP nick end labelling (TUNEL) in different stages of the tooth development of Sprague-Dawley rats. RESULTS: The positive cells of PCNA and PCD predominantly appeared at the proliferating growth center in the bud, the cap and the earlier bell stages of tooth development. PCNA and PCD were also observed in ameloblasts and odontoblasts, especially in the stellate reticulum cells of enamel organ and dental papilla during the period of dentin formation. CONCLUSION: Cell proliferation and programmed cell death are interrelated and interact on each other in the development of teeth, and they both involve in sculpturing the shape of teeth. PMID- 12539695 TI - [The influence of Millard-lip-repair on maxillary growth: an experiment study in rabbits]. AB - OBJECTIVE: To study the effects of the cheiloplasty(Millard rotation-advancement lip-repair) on the maxillary growth. METHODS: Thirty Japanese white rabbits were randomly divided into three groups, Group I (unoperated control group), Group II (unrepaired control group) and Group III (repaired with Millard-lip-repair). All the animals were sacrificed and measured on the clean slulls directly at 26 weeks postoperatively (30 weeks old). RESULTS: It was found that Group III (Millard repair group) had the anterioposterior growth disturbance of maxilla than Group I and II, while the height of maxilla and width of posterior dental arch were larger than that of the two control groups for compensation, and the nasal septum of Group III deviated to the cleft side. CONCLUSION: Millard-rotation-advancement lip-repair operation has inhibitory effects on maxillary growth. PMID- 12539696 TI - [An immunohistochemical and morphometric study of myoepithelial cells in salivary glands of human and experimental animals]. AB - OBJECTIVE: To investigate whether differences exist between myoepithelial cells of human beings and experimental animals. METHODS: Myoepithelial cells(MeCs) of 8 kinds of experimental animals were studied immunohistochemically using muscle acting-specific monoclonal antibody HHF35 and morphometrically using image analysis. RESULTS: There were differences in mean proportional volumes of MeCs between human beings and experimental animals. The mean proportional volumes of mucous acini were more than those of serous acini except salivary glands of rodents. CONCLUSION: The mean proportional volume of MeCs in salivary glands associates with animal species and secretion of salivary glands. PMID- 12539697 TI - [Preparation of Streptococcus sanguis DIG-labeled DNA probes and clinical application]. AB - OBJECTIVE: To investigate oral Streptococci nif gene. METHODS: According to the 800 base pairs(bp) sequence, a couple of special prime was designed, which were 5'TAGTTGAAGCGATTAAAGCG 3' and 5' TCGTCGTGAAAGCCAAA 3'. DIG-labeled DNA probes of ATCC10556 800 bp DNA fragment and 559 bp DNA fragment were made. By using Southern blotting and dot blotting, more than 30 bacterial strains were checked. RESULTS: 559 bp sequence was according with the reported results, and the Southern blot results showed that S34 sr, S34 no 1 strains had homologicous sequence with ATCC10556, two strains of Streptococcus sanguis were weak positive, and other strains were negative. CONCLUSION: Streptococcus sanguis ATCC10556 strain contained exactly nif gene and nif gene may be the special gene of Streptococcus sanguis ATCC 10556. PMID- 12539698 TI - [Cartilage matrix synthetic ability of rabbit articular chondrocytes cultured in vitro]. AB - OBJECTIVE: To observe the ability of matrix synthesis of rabbit articular chondrocytes cultured in vitro. METHODS: Rabbit articular chondrocytes were acquired by digestion of 0.1% type II collagenase and cultured in vitro, then cell morphology and ultrastructure were observed with phase-contrast microscope and scanning electronic microscope. With continuous culturing of chondrocytes in DMEM medium containing 15% FBS, synthesis of cartilage matrix was observed by using microscope and safranine-O staining. RESULTS: The cultured chondrocytes were polygonal cells. There were many rough endoplasmic reticula and mitochondria in cytoplasm, and a lot of secretory vesicles under cell membrane and in the cytoplasm. When cultured for 10 days, some small and white nodules were formed on the bottom of the culture dishes, and volcanic-mouth-like structures were formed when cultured for 20 days. Both these nodules and structures contained GAG positive substances were demonstrated by safranine-O staining. CONCLUSION: Chondrocytes can produce matrix and cartilage-like tissue in vitro, so it is feasible to produce cartilage by culturing chondrocytes in vitro. PMID- 12539699 TI - [Study of effects of six caries-preventive agents on enamel decalcification]. AB - OBJECTIVE: To compare the influence of 6 caries-preventive agents containing fluoride and trace element on dental enamel decalcification and observe the relationship between enamel decalcification and time of early carious lesions. METHODS: 84 bovine teeth were selected and divided into 7 groups. Experimental groups were treated with SrF, SnF2, (NH4)2 MoO2F4, ZnF, LaF and NaF with the same [F-]concentration and deionised water respectively, then all the specimens formed artificial carious lesion in lactic acid gel. RESULTS: The [Ca2+] concentration of lactic acid gel showed a variety of enamel decalcification among specimens treated with fluoride agents after 48 hours, on the other hand, all the specimens presented a platform of enamel decalcification after 48 hours. SnF2 demonstrated the strongest inhibition on enamel decalcification, meanwhile, NaF has the minimum influence on it. CONCLUSION: The time of forming artificial carious lesion with acid etch is about 48 hours, moreover, caries-preventive agents containing fluoride and trace element are effectively able to inhibit enamel decalcification, and the potential mechanism is the interaction of fluoride and trace element. PMID- 12539700 TI - [Effect of lipopolysaccharide on recombinant human osteogenic protein-1-induced responses in human dental pulp cells]. AB - OBJECTIVE: To investigate the effects of lipopolysaccharide (LPS) on recombinant human osteogenic protein-1 (rhOP-1) induced proliferation and differentiation. METHODS: Measured MTT, alkaline phosphatase (ALPase) activity and osteocalcin content of human dental pulp cells. RESULTS: Low-dose LPS (1 microgram/ml) caused co-operative effect with low concentration of rhOP-1 to enhance human dental pulp cell proliferation and ALPase activity, but not to osteocalcin, whereas, high dose LPS (100 micrograms/ml) significantly inhibited the proliferation, ALPase activity and osteocalcin with or without rhOP-1. CONCLUSION: The role of LPS in caries-induced reparative dentin might be a significant determinant. PMID- 12539701 TI - [Evaluation of methods of drug release rate in root canal disinfection controlled release delivery system containing metronidazole]. AB - OBJECTIVE: To measure the release rate of drug in controlled-release delivery system containing metronidazole. METHODS: Drug release tests in vitro, in vivo and through the roots of extracted teeth were performed. The release rate of metronidazole was calculated after 1 day, 3 days, 7 days and 10 days, and the results were statistically analyzed. RESULTS: Significant difference was observed among the total drug release rate in vitro, in vivo and through the roots of extracted teeth (P < 0.01). There was no statistically significant difference between the release test in vivo and through the roots of extracted teeth in vitro (P > 0.05). CONCLUSION: The method of using extracted teeth and the controlled release medium at 37 degrees C, pH7.4 was close to the situation in vivo, and it was an effective method to measure the drug release rate of the root canal disinfection controlled-release delivery system. PMID- 12539702 TI - [Extract of cemental matrix enhance periodontal cells bind to the root surface]. AB - OBJECTIVE: To investigate whether the extract of cemental matrix can enhance periodontal cells binding to the root surfaces. METHODS: Healthy human gingiva and periodontal ligament were acquired from patients, then gingival fibroblasts and periodontal cells were cultured in vitro. On the other hand, extract of cemental matrix was separated and clarified from healthy teeth which were extracted because of orthodontic treatment. Finally, the effects of cementum matrix with different concentration and different time on the attachment of gingival fibroblast and periodontal cells to root surface were observed. RESULTS: The extract of cemental matrix could enhance the initial attachment of human gingival fibroblasts (HGF) and human periodontal cells (HPC) to the surface of root, and such effects were strengthened with increasing density and time. The optimal concentration of extract of cell attachment 10 micrograms/ml, and the attachment was also proportional to the incubation time, reaching near maximal levels at 2 hours, furthermore, the extract of cemental matrix was more effective in promoting the attachment of human periodontal cells than that of gingival fibroblasts. CONCLUSION: The extract of cemental matrix can enhance the initial attachment of HGF and HPC on the root surface. PMID- 12539703 TI - [Detection of neuroendocrine feature about salivary adenoid cystic carcinoma]. AB - OBJECTIVE: To study the possibility of neuroendocrine feature of salivary adenoid cystic carcinoma (SACC). METHODS: Immunohistochemical staining was operated in SACC with S-100 protein, Neuron-specific enolase (NSE), chromagranin A and synaptophysin which are specific and correlative with neuroendocrine and neoplasias. RESULTS: The positive expression of S-100, NSE, chromogranin and synaptophysin were respectively 42, 32, 8 and 7 in 50 cases of SACC. CONCLUSION: There is possibility of neuroendocrine feature in some SACC cells. PMID- 12539704 TI - [Rat submandibular gland tumor induced by 3-methylcholanthrene]. AB - OBJECTIVE: To study the genesis and the development of salivary gland tumor (SGT). METHODS: SGT animal models were established by injection of 3 methylcholanthrene (3-MCA) oil solution into 40 submandibular glands of Sprague Dawley (SD) rats, then histopathology and ultrastructure of induced tumors were observed under microscope, and immunohistochemical detection of keratin and actin was carried out. RESULTS: 32 models of submandibular gland tumors were induced successfully, with 4 rats of squamous cell carcinoma and 28 leiomyosarcoma, and the carcinogenesis of 3-MCA and tissue genesis of induced tumors were discussed. CONCLUSION: Submandibular gland tumor can be induced by 3-MCA, and sarcoma was of high incidence. PMID- 12539705 TI - [Detection of Epstein-Barr viral genome in tumor cells of Warthin's tumor of parotid gland]. AB - OBJECTIVE: To evaluate the relations between Warthin's tumors of salivary gland and Epstein-Barr virus (EBV). METHODS: 75 cases of salivary gland Warthin's tumor and 20 cases of normal salivary gland tissue were examined for to detect EBV genoma by using the polymerase chain reaction (PCR) of DNA, 62 cases of single Warthin's tumor and 13 cases of multiple/bilateral Warthin's tumor's. RESULTS: EBV DNA was detected in 13 cases of single Wathin's tumors, 9 cases of multiple/bilateral Warthin's tumor and 3 cases of normal salivary gland. CONCLUSION: There was a close relationship between EBV and multiple/bilateral Warthin's tumors. PMID- 12539706 TI - [Study of cleft palate with problems of middle ears and its treatment]. AB - OBJECTIVE: To gain a clear idea of the following: 1. the relationship between the incidence of functional obstruction of middle ear and surgery on cleft palate and patients' age; 2. the bacterial culture of secretion taken from the tympanic cavity with secretory otitis media; 3. the preliminary effects of treatment by inserting a tube into the tympanic cavity of a patient with secretory otitis media. METHODS: 275 middle ears of patients with cleft palate were selected, then acoustic immitence examination and hearing threshold of pure tone were adopted, and bacterial culture was carried out for studying secretion of the middle ear with secretory otitis media. RESULTS: The tympanograms of 75.5% ears of the patients before cleft palate operation were found to be abnormal, among which patients under 3 years were 81%, and 41.2% ears of those after closure of cleft palate were abnormal. Gram's positive and negative germs of bacterial culture were 20.1% and 24% respectively. The hearing of 6 ears with secretory otitis media has recovered from the moderate hearing loss due to tube insertion. And the tympanograms of 4/8 ears were transformed from type B to type A. CONCLUSION: It is beneficial to improve function of Eustachian tube if cleft palate can be repaired at an early stage and beneficial to improve the patients' hearing and devlope their speech if a tube is inserted into the typanic cavity in cases of secretory otitis media at an early stage. PMID- 12539707 TI - [Surgical reduction and rigid internal fixation of midface fractures]. AB - OBJECTIVE: To determine the cause, clinical characteristics and treatment of midface fractures. METHODS: A retrospective review of 98 patient records was conducted. Data regarding age, cause of fracture, anatomic site, treatment modalities and clinical results were reviewed. RESULTS: Of the 98 patients, 78.6% were male, with the peak incidence occurring in the age group from 21 to 40 years old. Of the fractures, 32.6% were due to traffic accidents, 24.5% to assaults, and 17.3% to accidental falls. Most patients (87.8%) were treated by open reduction surgery and rigid internal fixation. Only 12.2% of patients were treated by closed reduction fixation. Almost all of the patients regained normal face appearance and functional occlusion. CONCLUSION: Surgical reduction and rigid internal fixation of midface fractures is a satisfactory method of treatment. PMID- 12539708 TI - [The investigation on nitric oxide levels in saliva and their relationship with the severity of periodontitis]. AB - OBJECTIVE: To study nitric oxide levels in saliva of patients with periodontitis and the relationship between nitric oxide levels and the severity of periodontitis. METHODS: Samples of saliva from 23 adult subjects with periodontitis and 27 healthy control subjects were collected at their first appointment. Data of gingival bleeding index (GBI), pocket depth (PD) and attachment level (AL) were recorded. Salivary nitrite was detected with Greiss method, which stood for the content of nitric oxide (NO). RESULTS: It was found that NO in saliva of patients with periodontitis were significantly higher than that in saliva of healthy individuals (P < 0.01). The levels of NO were 55.361 +/ 13.319 mumol/L and 28.806 +/- 6.604 microns/L in saliva with periodontitis and control respectively. Significant relationship was found between AL and salivary NO level (P < 0.02) and between PD and salivary NO level (P < 0.05). CONCLUSION: The change of salivary NO level is related with periodontitis. PMID- 12539709 TI - [The value of diagnosis with technetium-99m sequential scintigraphy of parotid gland in Sjogren's syndrome]. AB - OBJECTIVE: To explore the value of diagnosis with 99mTcO4- sequential scintigraphy of parotid gland in Sjogren's syndrome. METHODS: Technetium 99m(99mTcO4-) sequential scintigraphy was used in 52 cases of Sjogren's syndrome. RESULTS: The results were analysed quantitatively with activity curve by using ROI computer program. 46 cases (88.46%) were parotid swelling. The contour was clear in 28 cases (53.85%) and blurred in 18 cases (34.62%). The function of parotid was normal in 7 cases (13.46%), parotids with light damage in 15 cases (28.84%), parotids with middle damage in 17 cases (32.69%), and parotids with serious damage in 13 cases (25%). Most cases of damaged secretary function was present in the early phase, and a few cases of damaged absorption function was found in the following process. CONCLUSION: 99mTcO4- sequential scintigraphy of parotid gland is greatly valuable to diagnosis of Sjogren's syndrome. PMID- 12539710 TI - [Observation on the immune function of circulating red cells of patients with periodontal diseases]. AB - OBJECTIVE: To find out the relation between the causes of periodontal diseases and the immune function of red blood cells (RBC). METHODS: The functional state of SOD activity, red blood cell C3b receptor rosstte (C3bR), red cell immune complex rosstte (RBCICR), RBC immune adherence enhance factor (RFER) and RBC immune adherence inhibitor factor (RFIR) of red cells from the systemic circulation of patients with periodontal diseases were determined and compared with the normal control group. RESULTS: The SOD activity C3bR and RFER of red cells were lower, while RFIR and RBCICR were higher for patients with periodontal diseases comparing to the normal control. CONCLUSION: It indicates that the compromised function of red cells may contribute to the initiation and development of periodontal diseases. PMID- 12539711 TI - [Color comparison of VITA shade guide and metal ceramic restoration]. AB - OBJECTIVE: To understand color distribution of Vita shade guide and metal ceramic restoration and make Vita shade guide more reasonable in clinical applying. METHODS: The international standard CIE1976L*a*b* color system and Minolta CR-100 spectrocolorimeter were selected to measure and compare the characteristics of the shade guide and metal ceramic restoration. RESULTS: Color differences were noted between the shade guide and ceramic restorations. The value of metal ceramic restoration was brighter than that of VITA shade guide tab. There was different chroma between metal ceramic restoration and Vita shade guide. Metal ceramic restorations were yellower and redder. But no saturation differences were found between ceramic restorations and D2, D3 tabs. CONCLUSION: During clinical color-matching of metal ceramic porcelains, it is suggested that a VITA shape guide tab with lower brightness and paler color than that of adjacent teeth should be selected. PMID- 12539712 TI - [Clinical effect observation of methods of mandibular fracture treatment]. AB - OBJECTIVE: To observe and search for an adaptative and efficient treatment methods of mandibular fracture. METHODS: 218 cases of mandibular fracture patients were treated with different methods from 1991, then the relationship between methods, and treatment efficiency was appraised objectively. RESULTS: Different ways had different advantages, and the total successful rate was 94.5%. Titanium miniplates was a promising method for mandibular fracture treatment. CONCLUSION: It's crucial to select a right method in the mandibular fracture treatment. PMID- 12539713 TI - [Quantitative evaluation of new bone formation in the posterior zone of the glenoid fossa in response to the mandibular advancement in rats]. AB - OBJECTIVE: To quantify the new bone formation in the glenoid fossa of the temporal bone in response to the mandibular advancement. METHODS: A bitejumping appliance with guide slope was fit on the upper jaw of each rat in the experimental groups, resulting in a downward and forward displacement of condyle and continuous mandibular advancement. Sections were cut through the glenoid fossa at the sagittal plane and stained with PAS under the same condition. The new bone formation at the posterior zone of the glenoid fossa was detected by quantitative measurement through computer-assisted image analyzing system (Q550IW, Leica) with the advanced software (Leica Qwin Pro, Version 2.2). Statistical analysis was performed with GraphPad 3.0. RESULTS: 1. a comparison among experimental groups revealed a peak of new bone deposition at the 21-day group, significantly greater than other groups (P < 0.001); 2. a comparison among control groups showed a gradual decrease of new bone deposition characterized by a peak of at the 3-day group and a bottom at the 30-day group (P < 0.001); 3. a comparison between experimental and control groups showed no significant difference statistically at the 3-day groups (P > 0.05), whereas a significant difference of new bone deposition starting from 7-day groups, with a peak at 21 day groups and a decrease towards 30-day groups (P < 0.001). CONCLUSION: The advancement of mandible leads to the remodeling of the glenoid fossa indicated by the new bone deposition in growing rat. PMID- 12539714 TI - [Image analysis of condylar cartilaginous adaptation to mandibular protrusion in rats]. AB - OBJECTIVE: To quantify chondrogenesis in condylar cartilage in response to active forward positioning of mandible. METHODS: Identical bite-jumping appliances were fixed to upper incisors of animals in experimental groups, causing a continuous mandibular forward positioning. Histological sections were performed through mandibular condyle and were stained with HE under the same condition. Chondrogenesis was quantified by measuring the area of cells in resting, proliferative, hypertrophic and erosive zones in the superioposterior region, through computer-assisted image analyzing system (Q550IW, Leica) with the advanced software (Leica Qwin Pro, V2.2). Statistical analysis was performed with GraphPad 3.0. RESULTS: 1. comparison among experimental groups revealed a fluctuation in proliferative zone, with 21-day group being the greatest (0.058 +/ 0.004 mm2) and 14-day the least (0.012 +/- 0.001 mm2). A change in erosive zone was also depicted by a peak of 0.112 +/- 0.001 mm2 in 7-day group and a bottom of 0.018 +/- 0.002 mm2 in 14-day group (P < 0.001). 2. comparison among the control groups manifested stable zonation, except for a slump of proliferative zone descending from 0.069 +/- 0.005 mm2 in 3-day group to 0.009 +/- 0.001 mm2 in 21 day group (P < 0.001). 3. comparison between experimental and control groups demonstrated significant discrepancy in proliferative zone, hypertrophic zone and erosive zone (P < 0.001). CONCLUSION: Mandibular forward positioning stimulates and accelerates cartilaginous remodeling in mandibular condyle. PMID- 12539715 TI - [Determination of normal temperature properties of refractory die material compatible with slip casting core of sintered titanium powder]. AB - OBJECTIVE: The refractory die is the precondition for developing slip casting core of sintered powder. This study is to determine the normal temperature properties of the refractory die material compatible with slip casting core. METHODS: to mix the die material at five different ratios (8/1, 7.5/1, 7/1, 6.5/1, and 6/1) and measure their solidification time with self-manufactured Vicker's needle; to prepare five cylindrical specimens (phi 10 x 15 mm) in different drying time for determining their compressive strength, and then to let another five specimens fire at 1000 degrees C four times for measuring the residual compressive strength at room temperature. RESULTS: The setting time was 16.25 minutes (7.5/1), and the lower the powder-liquid ratio, the longer the setting time. The normal compressive strength was 25.32 MPa (drying 24 hours), while the longer the drying time, the higher the compressive strength achieved (P < 0.01). The residual compressive strength was 24 MPa. CONCLUSION: The normal temperature properties of the refractory die material meet the demand of slip casting core of sintered powder. PMID- 12539716 TI - [Determination of high temperature compressive strength and refractory degree of die material compatible with slip casting core of sintered titanium powder]. AB - OBJECTIVE: The refractory die is the precondition for developing slip casting core of sintered powder. This study is to determine the high temperature properties of the refractory die material compatible with slip casting core. METHODS: To prepare three cylindrical specimens (phi 10 x 15 mm) and determine their compressive strength at 1000 degrees C: to make four specimens in flat topped cone for determining the practical refractory degree by decreasing the pressing temperatures in a sequence of 1420, 1400, 1350 and 1100 degrees C. RESULTS: The compressive strength of this material was 17.8 MPa at 1000 degrees C. Its practical refractory degree was higher than 1100 degrees C. CONCLUSION: The high temperature properties of the refractory die material that we developed meet the demand of slip casting core of sintered powder. PMID- 12539717 TI - [A study on biological properties of fimbriae of Actinomyces viscosus. II. Chemical characteristics of fimbriae of A. viscosus]. AB - OBJECTIVE: To investigate the chemical characteristics of two purified fimbriae. METHODS: The specific antibodies against type I and type II fimbriae of A. viscosus were prepared and the purity of the two purified fimbriae had been identified by the specific antibodies. Then, chemical analysis and amino acid analysis of the two purified fimbriae were done. RESULTS: The two purified fimbriae were mainly composed of proteins and small amount of carbohydrate. The two types of fimbriae contained large amounts of aspartic acid, glutamic acid, alanine, lysine and glycine, and high percentage of nonpolar amino acid was analysed while basic amino acids were present in a very small amounts. CONCLUSION: These results may help to understand the relationship between the chemical characteristics of two types of fimbriae and their biological activity. PMID- 12539718 TI - [A study on biological properties of fimbriae of A. viscosus. III. Adherence activity of fimbriae of A. viscosus]. AB - OBJECTIVE: To investigate the adherence activity of two types of fimbriae of A. viscosus on tooth surface or with S. sanguis 34. METHODS: The inhibited adherence tests, coaggregation tests and the inhibited coaggregation tests were done. RESULTS: 1. The purified type I and type II fimbriae inhibited the adherence of A. viscosus to salivary-treated hydroxyapatite (SHA) and the two specific IgG to type I and type II fimbriae blocked the adsorption of strain T14V, strain 5519 and strain 5951 to SHA; 2. Only type II fimbria indirectly mediated the visible agglutination of S. sanguis 34 and only IgG to type II fimbriae inhibited coaggregation of strain T14V and strain 5915 with S. sanguis 34. CONCLUSION: Type I and type II fimbriae have adherence ability, and only type II fimbria has the agglutination activity. Additionally, the methods which were used to prepare fimbriae don't damage the biological activity of fimbriae. PMID- 12539719 TI - [Studies on standard growth curve chart on body height of Chinese adolescent for orthodontic diagnosis]. AB - OBJECTIVE: To make a standard growth curve chart on the body height of Chinese adolescent. METHODS: Statistical data were surveyed in 1995, which were obtained from subjects consisting of 32,524 students aged from seven to nineteen years in the province of Sichuan. RESULTS: Mean values of the annual increment of body height and the ages of the pubertal growth periods were analyzed. CONCLUSION: The new standard growth curve chart on the body height of adolescent chinese were applied for better understanding the growth pattern of chinese orthodontic patients. PMID- 12539720 TI - [An experimental study on demineralization of several beverages on bovine enamel]. AB - OBJECTIVE: The aim of this study is to investigate the demineralization effects of several beverages on bovine enamel. METHODS: The concentration of calcium and phosphate in these beverages were measured by using EL 312e Micro-Plate after the bovine enamel was exposed to beverages for a period of 7 days. RESULTS: The data indicated that almost all of these beverages could cause demineralization of bovine enamel, except mineral water, and the concentration of calcium and phosphate in these beverages changed after the bovine enamel was exposed to beverages. There were great significant differences in demineralization abilities among different beverages (P < 0.01). The concentration of calcium and phosphate in beverages also changed with time (P < 0.01). CONCLUSION: All these beverages, except mineral water have erosive effects on bovine enamel. Demineralization degree varied with kinds of beverages. The fruit juice has the highest erosive effect on the enamel, and the calcium milk has the lowest erosive effect on the bovine enamel, when comparing with other beverages. PMID- 12539721 TI - [A comparative study about the influence of lanthanum, calcium, fluoride on plaque pH]. AB - OBJECTIVE: The study is designed to compare the effects of Lanthanum (La), Calcium (Ca), Fluoride (F) on pH changes of plaque, and to discuss the anti cariogenic mechanisms of Lanthanum in terms of metabolism. METHODS: Plaque pH values after sucrose solution (containing La3+, Ca2+, F-) rinse were measured contiguously using micro-touch method, and then the minimum pH, time of minimum pH, ranges of plaque pH (delta pH), and areas of the response curves below (AUC6.5) were analyzed using two-way analysis of variance (ANOVA). RESULTS: No statistical difference was found in the above monitored parameters of 8 subjects. For the decreasing degree of plaque pH, the pH value in the control group fell down the most obvious, and the second was that in the Ca group. The pH value decreased the least in the F and La group, and there was no significant difference between the F and La group. The time of minimum pH was the same in each group. CONCLUSION: Ca, La, F can decrease the range of plaque pH drop after sucrose solution rinse. The degree of Ca was the weakest, and F and La were same. All the above evidences prove that La may inhibit the acidogenicity of plaque. PMID- 12539722 TI - [The examination of DNA repair capacity in the patients with oral precancerous lesion]. AB - OBJECTIVE: The occurrence of oral precancerous lesion is related with not only carcinogenic environmental factors but also some self-regulating mechanisms. The genetic heredity factors determine individual's DNA repair capacity. The study is attempted to investigate the change of DNA repair capacity of the patients with the precancerous lesion. METHODS: The study is conducted in 16 pairs of oral precancerous lesion and normal controls by using host cell reactivation assay, while some dangerous factors such as smoking, alcohol intaking and family history of cancers are also recorded. RESULTS: The host cell reactivation in the patients with precancerous lesion is lower than that of normal controls (P < 0.05) and is shown to be an independent risk factor in the occurrence of oral precancerous lesion (OR: 3.25 95% CI = 1.01-10.49). CONCLUSION: The host cell reactivation is likely to be a helpful indicator for predict prognosis of cancer for the individuals with highly genetic sensitivity to cancer. PMID- 12539723 TI - [The effects of MMPs and TIMPs on the metastasis of oral squamous cell carcinoma to neck lymph nodes]. AB - OBJECTIVE: This study is aim at investigating the relationship between the expression and activity of MMP-2,9, TIMP-1,2 and the metastasis of oral squamous cell carcinoma to neck lymph node. METHODS: The expression of MMP-2,9, TIMP-1,2 in 30 oral squamous cell carcinoma, two oral squamous cell carcinoma cell lines GNM and TSCCa was detected by in situ hybridization; Zomography was performed to determined the activity of MMP-2,9 in cancer tissue and conditioned medium of GNM and TSCCa; Ability of invasion of GNM and TSCCa was measured with in vitro invasion assays. RESULTS: Both tumor cells and stroma expressed both MMP-2,9 and TIMP-1,2; It was found that MMP-2,9, TIMP-1,2 had a higher expression in patients with neck lymph node metastasis than those without metastasis (P < 0.05), positive percentage of MMP-2,9 in GNM was higher than that in TSCCa; The MMP-2,9 in node-positive patients was more active than in node-negative ones (P < 0.05); GNM had more active MMP-2, 9 and higher invasive potential than TSCCa. CONCLUSION: MMP-2,9 may play an important role in metastasis of oral squamous cell carcinoma; Elevation of TIMP-1,2 is likely to be the result of interaction of tumor and stroma. PMID- 12539724 TI - [The isolation and identification of pathogenic bacteria from rampant caries in children]. AB - OBJECTIVE: In order to prevent rampant caries in children, the predominant pathogenic bacteria of rampant caries in children were defined. METHODS: The pathogenic bacteria of rampant caries were isolated and identified in 30 children with rampant caries, aged from 2 to 5 years old. Samples were collected from the caries of labial surfaces of maxillary anterior deciduous teeth, including caries lesions and caries-free smooth enamel surfaces. Plaque were obtained from the cervical third areas of the maxillary labial anterior deciduous tooth surfaces in children without rampant caries and caries-free children and served as controls respectively. The isolates were identified by colony morphology, biochemical tests and DNA base contents (mol percent G + C). RESULTS: The prevalence of Streptococcus mutans (S. mutans) and Streptococcus sobrinus (S. sobrinus) in the plaque of caries lesions and the proportions of S. mutans and S. sobrinus in the plaque of 2 sampling sites of children with rampant caries were all significantly higher than those of children without rampant caries and caries-free children (P < 0.05). CONCLUSION: S. mutans and S. sobrinus are predominant pathogenic bacteria in rampant caries in children. PMID- 12539725 TI - [Effects of different disinfectants on levels of endotoxin in canals of chronic periapical periodontitis]. AB - OBJECTIVE: The objective of this study is to compare effects of formocresol, camphorated phenol and Ya Kan (metronedazole) on endotoxin levels in canals of chronic periapical periodontitis. METHODS: A total of 30 patients with chronic periapical periodontitis were selected from outpatients, and 10 cases each were randomly picked out to treat by sealing with formocresol, camphorated phenol and Ya Kan respectively. All patients were both sampled before and after sealing of these disinfectants, and chromogenic substrate method of limulus amebocyte lysate (LAL) test was used to measure levels of endotoxin. RESULTS: The effects of Ya Kan, formocresol and camphorated phenol on levels of endotoxin did not showed significant difference. CONCLUSION: Some anti-anaerobic bacterial drugs can be used to treat chronic periapical periodontitis. PMID- 12539726 TI - [Effects of unilateral cleft lip and palate prosthesis on the development of maxillary and facial soft tissue]. AB - OBJECTIVE: The effect of unilateral cleft lip and palate prosthesis on the development of maxillary and facial soft tissue was investigated. METHODS: The standard cephalometric measurement was used in patients with complete unilateral cleft lip and palate operated in different periods. We obtained data by measurement. RESULTS: Hard tissue: ANS-X, PNS-X and N-ANS in the group of patients were statistically different from those of the normal control group, but in ANS-PNS and N-PO, there is no statistical difference in these two groups. Angles of hard tissue: There is statistical difference in < N. W. ANS and < SNA between the control and the group of patients. There is only statistical difference in < SNB between the D group and the control group. There is no difference among other groups. Ratios of hard tissue: The ratios of ANS-X/S-X and PNS-X/S-x in the control group were higher than those of the group of patients, and the ratios of S-Go/N-Po, Ar-Go/N-Po and N-Po/AN in the group of patients were higher than those of the control group. The ratio of NMC/SUBN: It is the highest ratio in the control group, and there is no difference between ratios in the control group and the C group, but there is statistical difference between the control and other patient groups (P < 0.05). CONCLUSION: Comparing with normal persons, the defect of maxillary length development is apparent in the patients with complete unilateral cleft lip and palate due to congenital development deformity. The figure of soft tissue in patients with unilateral cleft lip and palate is anastomotic with the figure of their hard tissue. The excavation of bony middle 1/3 in face results in paramorphia of soft tissue adhering on the bony tissue. We can conclude that in order to obtain better articulation function, the palatoplasty should be carried out early. PMID- 12539727 TI - [Inhibitive effects of lip repair on maxillary growth in patients with complete unilateral cleft lip and palate]. AB - OBJECTIVE: The aim of this study was to assess the isolated effects of lip repair on inhibition of maxillary growth in patients with complete unilateral cleft lip and palate. METHODS: The lateral cephalometric analysis were applied to 20 patients with unilateral cleft lip and palate who only had lip repaired in childhood, 32 patients with unilateral cleft lip and palate who had both lip and palate repaired in childhood, as well as 37 normal Chinese of the same age as controls. RESULTS: Both complete unilateral cleft lip and palate groups had almost the same significant degree of maxillary retrusion compared with the normal control group. CONCLUSION: Lip repair is one of important factors that could inhibit maxillary growth in patients with complete unilateral cleft lip and palate. PMID- 12539728 TI - [Antibacterial activities of seven commonly used antibiotics to 106 isolated strains of porphyromonas gingivalis in vitro]. AB - OBJECTIVE: Porphyromonas gingivalis (Pg) is one of the main putative pathogens of periodontal disease, especially of adult periodontitis. This in vitro study was aimed at observing the inhibitory activities of 7 commonly used antimicrobical drugs on 106 stains of Pg which isolated from subgingival plaques of sites with adult periodontits. METHODS: Kirby-Bauer paper disk method was used in this study. RESULTS: Pg was highly susceptible to Clindamycin, Sulfamzthoxazole timethoprin, Tetracycline and Cefazolin, the susceptibility rates were 93.4%, 78.3%, 60.4% and 56.6% respectively. CONCLUSION: Majority of the strains of Pg was moderately susceptible to cefalexin and acetylspiramycin, while no strains of Pg was susceptible to Penicillin G. The concentrations of Pg solutions (10(5) CFU/ml, 10(6) CFU/ml and 10(8) CFU/ml) had not influence on the results. This data is helpful for clinically antibiotic selection of periodontal treatment. PMID- 12539729 TI - [Development and applications of a computer aided complete denture design system]. AB - OBJECTIVE: The purpose of this study was to develop a practical CAD/CAM system with knowledge databases for complete denture. METHODS: A three-dimensional coordinate measuring machine (3DCMM) was used to collect three-dimensional information of edentulous models and bite plates and the mathematics model of complete denture was established using B-Spline method. This system was established by seting eight functional models: the measuring model, the converting data model, the constructing curve-surface model, the producing base and the editing model, the arranging artificial and the editing model, the outputting income model, the imitating dynamic model and the managing model. RESULTS: (1) The soft and hard system of a computer aided complete denture design (CACDD) was established, including 3DCMM, 8 functional models and databases. (2) A CACDD system and the three-dimensional demonstration were completed. CONCLUSION: This system lay down the foundation for setting up the computer aided complete denture design/manufacturing system. PMID- 12539730 TI - [Changes of soft tissue profile in operated unilateral cleft lip and palate patients after maxillary protraction]. AB - OBJECTIVE: The aim of this study is to investigate effects of maxillary protraction on soft tissue profile in operated operated unilateral cleft lip and palate (UCLP) patients. METHODS: A total of 10 growing UCLP patients (male 7, female 3), age from 8.2 to 12 years old (Average: 10.4 years old), were selected to be treated with maxillary protraction using head gear-chin cap-long hook protraction appliance. The appliance was worn 12-14 hours per day, and the protraction force was 400-500 g each side. The protraction direction was forward and slightly downward. The treatment period was 4.7 months (Average: 5.8 months). Cephalometrics were taken before and after treatment. The changes of soft-tissue profile were studied using the computer-aid X-ray cephalometric analysis. RESULTS: After protraction, the points of Prn, Sn and Ls moved forward significantly. The distance from points Ls to E plane changed significantly from 0.46 mm before treatment to 1.18 mm after treatment. The angle G-Prn-Pg' decreased significantly, and G-Sn-Pg' changed significantly from -0.30 before treatment to 6.260 after treatment. The anterior-posterior position of mandible and lower lip did not change significantly, the changes of angles Cm-Sn-Ls, A'ls/SiLi had no statistical significance. The results indicated that maxillary protraction could make maxilla and upper lip move forward, and the convexity of soft tissue profile improve significantly. CONCLUSION: Maxillary protraction is an effective way to improve the facial deformity of operated UCLP patients. UCLP patients should have early interrupted treatment. PMID- 12539731 TI - [Clinical significance of trigeminal neuralgia treated using radiofrequency thermocoagulation (RFT) with different approaches]. AB - OBJECTIVE: The authors presented the short term and long term effects of the trigeminal neuralgia treated by RFT with anterior-lateral facial, lateral facial and trans-subzygomatic approaches. METHODS: 320 primary patients with trigeminal neuralgia were selected for this study. All these patients were followed up with the period of 1 to 6 years. RESULTS: The first-time effective ratios of pain relieve treated by using the anterior-lateral facial approach, and the lateral facial and the trans-subzygomatic, were 93.3% and 73.3% respectively. Most of the patients (92.5%) achieved excellent or good pain-relieve after treated by one of these three approaches. Compared with the lateral facial and trans-subzygomatic procedures, the anterior-lateral facial approach caused less recurrent rate in long term effects (P < 0.01). The late recurrence was found in 93 patients, but pain-relieve effect still could be obtained by retreating with the former method and without difficulty or hazard. One patient had serious Rollets syndrome. CONCLUSION: It is concluded that the anterior-lateral approach is a method with more effective pain-relieve and less pain recurrence rate, and the trans subzygomatic method can be used as a substitutive approach in the treatment of the second branch of trigeminal neuralgia. PMID- 12539733 TI - [The influence of the hamulotome and interplatal incision on the middle ear function]. AB - OBJECTIVE: The aim of this study is to explore the effect of hamulotome on the eustachian tube function and verify the function of the tensor veli palatili muscle on eustachian tube. METHODS: Single-sided hamulotome were carried out in the rabbits under the direct visual. And the middle ear cavity mucosa of the rabbits was collected at different intervals and dyed with PAS and PH to observe the middle ear changes under microscope. RESULTS: Middle ear function changed obviously after the hamulotome, with increase in globular cells, submucosal edema and inflammatory cells. And mucin was not found in several cavities. CONCLUSION: Hamulotome do affect on the middle ear function and should not be damaged in order to protect the middle ear function in some patients. PMID- 12539732 TI - [The influence of palatoplasty and tympanotomy on middle ear function in cleft palate patients]. AB - OBJECTIVE: The aim of this study is to explore the influence of palatoplasty and tympanotomy on middle ear function or hearing condition in cleft palate patients. METHODS: A total of 22 ears with otitis media with effusion in cleft palate patients were inserted with PE tubes, and Pre- and post-operative hearing level and middle ear function in these patients were tested and compared respectively, in order to demonstrate the action of tympanotomy and PE insertion on hearing and middle ear function. Another 38 patients with cleft palate were operated in their early age with the proper operation method, and pre- and post-operative middle ear function were examined and compared to show the influence of the operation on the middle ear function. RESULTS: PE Tube insertion and tympanotomy changed the pressure condition of middle ear cavity and raised the hearing level about 17 dB in the middle-ear-diseased cleft palate patients, and patients with palatoplasty did not show obvious changes in middle ear function in the short-term trail. CONCLUSION: Early and active tympanotomy do help in improving the function of the middle ear, and it is suggested to improve patients' hearing and middle ear function as early as possible. Palatoplasty did not affect the function of middle ear. PMID- 12539734 TI - [Construction of eukaryotic expression plasmid pcDNA3-gtfB expressing glucosyltransferase B of Streptococcus mutans]. AB - OBJECTIVE: Glucosyltransferase (GTF) of Streptococcus mutans is considered as a cariogenic virulence factor due to its ability to synthesize glucan, which facilitate sucrose-depended adherence and cell-to-cell accumulation of bacteria. In this study, gtfB, the target gene fragment which encodes multiple catalytic sites and antigen epitopes of GTF, was recombined into eukaryotic expression vector pcDNA3. The feasibility of the recombination plasmid pcDNA3-gtfB used as gene vaccine will be investigated in further study. METHODS: The target gene fragment gtfB (904-4578 bp) was obtained by standard PCR amplification while genome DNA of streptococcus mutans GS-5 was used as template. Then the PCR products were extracted and purified from low-melting temperature agarose. The gtfB and plasmid pcDNA3 were cut by Kpn I, Xho I, and the digested products were extracted and purified again for recombination. The purified gtfB and plasmid pcDNA3 were recombined by T4 DNA ligase, ligation products were transformed into competent cell, Escherichia coli JM109. Transformed colonies were screened by Ampr LB plate, then recombined plasmids were isolated and identified by restricted endonuclease cutting and Sanger dideoxy DNA sequencing. RESULTS: Identified by agarose gel electrophoresis, the target gene-gtfB obtained PCR amplification had the same molecular size (36 kb) as predicted. It was indicated that recombined plasmids contained inserted gtfB gene fragment by restricted endonuclease cut analysis, the sequencing data also indicated that inserted gtfB gene had correct DNA sequence and orientation according to DNA sequence of Streptococcus mutans GS-5 (gene bank M17361). CONCLUSION: Inserted gene-gtfB of recombined plasmid pcDNA3-gtfB encoded multiple catalytic sites and epitopes. It was proved that these epitopes had high immune antigenicity and that antiserum could significantly inhibit the synthesis of water-insoluble glucans and water soluble glucan. In vitro adherence experiment also indicated that it could inhibit streptococcus mutans adherence to saliva-coated hydroxyapatite. Vector pcDNA3 was high expressing eukaryotic vector, and could stimulate antigen representing cell. It was suggested that recombined plasmid pcDNA3-gtfB had high immune antigenicity and immune responsiveness, and this supported its use as gene vaccine candidates in the development of anti-caries vaccines. PMID- 12539735 TI - [The role of Fos protein in modulation of dental pain in central nerve system]. AB - OBJECTIVE: The aims of this study were to reveal the relationship between Fos protein expression, the transcription of preproenkephalin (PENK) mRNA, and the change of enkephalin (ENK) level initiated by experimental dental pain, and to understand the central modulation mechanism of dental pain. METHODS: The techniques of Immunohistochemistry in situ hybridization and radioimmunoassay were applied respectively to detect the expression of Fos, the transcription of PENK mRNA, and the change of ENK level in the experimental nucleus of the spinal tract of the trigeminal nerve, caudal part (sp5c) of rats. RESULTS: It was found that Fos protein expression was in a time-dependent manner. Fos appeared half hour after the pain stimulation, and its peak occurred two hours after the pain stimulation, but disappeared slowly four hours later. The transcription of PENK mRNA was seen two hours after the pain stimulation, its peak appeared four hours later and it disappeared eight hours later. The level of ENK increased significantly (P < 0.01) in the caudal part of the spinal tract of the trigeminal nerve four hours after pain stimulation. CONCLUSION: Fos protein may take a part in the central modulation of dental pain by initiating PENK mRNA transcription which led to the increment of ENK. PMID- 12539736 TI - [An experimental study on the blood vessel sclerosing therapeutic agents for cavernous hemangiomas]. AB - OBJECTIVE: This study was designed to test what are the suitable drugs for sclerotherapy and investigate the formation mechanism of cavernous hemangiomas. METHODS: A total of 84 rabbits were selected for this study, and divided into 6 groups, including one control group and five experimental groups. We selected auricular veins of rabbits as the location of injection. The rabbits of the control were injected with physiological saline, and the rabbits of the experimental groups were injected respectively with various drugs, including 5% sodium morrhuate, 10% alum, pinyungmycin, leurocristine and the combination of pinyungmycin and dexamethasone. Histopathological changes were analyzed after 3, 7, 14, 21, 28, 35 days of sclerotherapy. RESULTS: According to histopathological changes of tissue, 5% sodium morrhuate, 10% alum, and the combination of pinyungmycin and dexamethasone produced better eutherapeutic effects, when comparing with other methods. However, 5% sodium morrhuate caused tissue necrosis of the injection site. CONCLUSION: The combined therapy of pinyungmycin and dexamethasone produced excellent sclerotic effects. Whatever drugs causing serious damage of HVEC may contribute to the therapy for cavernous hemangiomas. PMID- 12539737 TI - WHO to hold urgent expert consultation on acrylamide in food after findings of Swedish National Food Administration. PMID- 12539738 TI - Monitoring of the dietary exposure of the population of chemical substances in the Czech Republic: design and history. AB - The basic objective of the programme of monitoring the dietary exposure in the Czech Republic is to describe the character of the health risk based on the exposure of the Czech population to chemical substances in food. The conception of the monitoring programme is based on the methods of the Total Diet Study, established on the principle of defining the consumption of the most important foodstuffs, their purchase in the network of stores in selected shops on fixed dates, their transport to the central laboratory, cooking and consequent chemical analyses in the central laboratories. In the Czech Republic monitoring has been conducted since 1991/1992 and routine activities have been ongoing since 1994. The programme is fully sponsored by the government. Monitoring carried out in 1994-2001 provided a considerable amount of information, which was applied when drawing programmes of health protection and promotion, in regulatory work and when solving problems of the international food trade. PMID- 12539739 TI - Post-exposure prophylaxis of HIV infection: German-Austrian recommendations. PMID- 12539740 TI - Opsin cDNA sequences of a UV and green rhodopsin of the satyrine butterfly Bicyclus anynana. AB - The cDNAs of an ultraviolet (UV) and long-wavelength (LW) (green) absorbing rhodopsin of the bush brown Bicyclus anynana were partially identified. The UV sequence, encoding 377 amino acids, is 76-79% identical to the UV sequences of the papilionids Papilio glaucus and Papilio xuthus and the moth Manduca sexta. A dendrogram derived from aligning the amino acid sequences reveals an equidistant position of Bicyclus between Papilio and Manduca. The sequence of the green opsin cDNA fragment, which encodes 242 amino acids, represents six of the seven transmembrane regions. At the amino acid level, this fragment is more than 80% identical to the corresponding LW opsin sequences of Dryas, Heliconius, Papilio (rhodopsin 2) and Manduca. Whereas three LW absorbing rhodopsins were identified in the papilionid butterflies, only one green opsin was found in B. anynana. PMID- 12539741 TI - Secondary acute myelogenous leukemia with MLL gene rearrangement following radioimmunotherapy (RAIT) for non-Hodgkin's lymphoma. AB - Targeted therapy with conjugated and unconjugated monoclonal antibodies for non Hodgkin's lymphoma has revolutionized the approach to this disease. The efficacy and low toxicity of these agents have allowed introduction of this strategy in the early stages of therapy. Longer follow-up is needed before validating the safety of these agents. Since monoclonal antibodies are being given as front-line therapy, it is important to identify all potential adverse events. We report a case of secondary acute myelogenous leukemia (AML) with 11q23 cytogenetic abnormality and mixed lymphoid leukemia (MLL) gene expression in a patient treated with Y90 labeled anti-CD20 antibody (Zevalin). The patient was not exposed to topoisomerase II inhibitors. Our observations suggest a relationship between 11q23 leukemia and radioimmunotherapy (RAIT) and further studies are needed. PMID- 12539742 TI - People's Charter for Health. People's Health Assembly. PMID- 12539743 TI - Ashworthius Patriciapilittae n. sp. (Trichostrongyloidea: Haemonchinae), an abomasal nematode in Odocoileus virginianus from Costa Rica, and a new record for species of the genus in the Western Hemisphere. AB - A species of Ashworthius is reported for the first time in the Western Hemisphere, and A. patriciapilittae n. sp. is described on the basis of specimens in white-tailed deer Odocoileus virginianus from Costa Rica. Among 8 known species, A. patriciapilittae is morphologically similar to A. tuyenquangi in red muntjac Muntjacas muntjak from northern Vietnam. The synlophe in A. patriciapilittae is composed of 26 ridges in the cervical zone and is continuous to the caudal extremity in males and females. Males are characterized by a complex dorsal ray and narrow trifurcate spicules (351-356 microm long) lacking an "eyelet." with dissimilar ventral and dorsal processes; the gubernaculum is 45 48% of the spicule length. Females have a prominent linguiform flap at the vulva and large eggs (108-142 microm long). The presence of A. patriciapilittae in Costa Rica is examined in the context of competing hypotheses for cospeciation or contemporary host-switching in cervids: either A. patriciapilittae is a component of an endemic Central and South American fauna that has diversified through coevolution of Ashworthius and cervid hosts or it has been introduced. Among haemonchines in the Western Hemisphere, specimens of A. patriciapilittae may be confused with 3 species of Haemonchus, including H. contortus, H. placei, and H. similis, that occur in both domestic and wild ruminants. Discovery of A. patriciapilittae emphasizes the continued need for survey and inventory to define the structure and distribution of parasite faunas in wild and domestic ruminants from the Nearctic and Neotropical regions. PMID- 12539744 TI - High prevalence of viable Toxoplasma gondii infection in market weight pigs from a farm in Massachusetts. AB - The ingestion of uncooked infected meat is considered important in the epidemiology of Toxoplasma gondii infection in humans and little is known of the prevalence of viable T. gondii in meat used for human consumption in the United States. In the present study, viable T. gondii was isolated from 51 out of 55 pigs destined for human consumption. Hearts and tongues (500 g) from fifty-five 6 mo-old pigs from a farm in Massachusetts were bioassayed for T. gondii by feeding them to T. gondii-free cats. Feces of these cats were examined for shedding of T. gondii oocysts. Fifty-one of 55 cats fed pig tissues each shed 25-810 million T. gondii oocysts in their feces. Two of these cats consumed tissues of pigs that were shown to be seronegative with the Sabin-Feldman dye test, the modified agglutination test, and the Western blot. Results indicate that until examination of meat for T. gondii infection is implemented in slaughterhouses, all meat should be cooked according to industry guidelines before human consumption. PMID- 12539745 TI - Porcine enteritis associated with Eimeria spinosa Henry, 1931 infection. AB - Coccidia of the genus Eimeria are present in most pigs raised on dirt in the United States. They are generally considered nonpathogenic in weaned pigs. Oocysts of Eimeria spinosa Henry, 1931 were observed in tissue sections and intestinal contents of a weaned male pig that died suddenly on a farm in Iowa. Microscopically, necrotizing enteritis associated with many thick-walled coccidial oocysts was present in intestinal sections. Examination of intestinal contents demonstrated oocysts that were thick-walled and had small projections on the surface of the oocyst wall, characteristic of E. spinasa Henry, 1931 of swine. Twenty-live oocysts in intestinal contents measured 20.4 by 14.2 microm. No pathogenic bacteria were detected in the pig by culture methods, but lesions suggestive of salmonellosis were observed in some tissues. The specific cause of death was not determined; however, E. spinosa infection was considered to have contributed to the death of this pig. The results suggest that E. spinosa may be pathogenic for pigs. PMID- 12539746 TI - [Conjunctival mixed injection with membrane development. Ocular involvement in epidermolysis bullosa hereditaria generalisata atrophicans]. PMID- 12539747 TI - A hypothetical reconstruction unveiling a family secret. PMID- 12539748 TI - [Recurrent bilateral corneal erosions and opacities in corneal stroma. Pre Descemet dystrophy in X chromosome recessive ichthyosis]. PMID- 12539749 TI - Quiz. Rectal mucocele. PMID- 12539750 TI - Getting under your skin. PMID- 12539751 TI - [Hypertrichosis lanuginosa of unknown origin in the area of the face. Porphyria cutanea tarda]. PMID- 12539752 TI - Chromosome instability of HPRT-mutant subclones induced by ionising radiation of various LET. AB - The induction of HPRT-mutations and survival of Chinese hamster cells (line B11ii FAF28, clone 431) were studied after irradiation by 4He and 12C-ions of various LET (20-360 keV/micrometers), produced by the U-200 heavy ion accelerator. The RBE increases with LET up to the maximum at 100-200 keV/micrometers and then decreases. Cytogenetic analysis was performed on the HPRT-mutant subclones selected from unirradiated Chinese hamster V-79 cells and from HPRT-mutant subclones that arose after exposure to gamma-rays, 1 GeV protons and 14N-ions (LET-77 keV/micrometers), produced by the synchrophasotron and the U-400M heavy ion accelerator. Slow growing mutant subclones were observed. The cytogenetic properties of individual clones were highly heterogeneous and chromosome instability was observed in both spontaneous and radiation-induced mutants. Chromosome instability was highest among spontaneous mutants and decreased with increasing LET. PMID- 12539753 TI - Chromosome aberrations in human lymphocytes induced by 250 MeV protons: effects of dose, dose rate and shielding. AB - Although the space radiation environment consists predominantly of energetic protons, astronauts inside a spacecraft are chronically exposed to both primary particles as well as secondary particles that are generated when the primary particles penetrate the spacecraft shielding. Secondary neutrons and secondary charged particles can have an LET value that is greater than the primary protons and, therefore, produce a higher relative biological effectiveness (RBE). Using the accelerator facility at Loma Linda University, we exposed human lymphocytes in vitro to 250 MeV protons with doses ranging from 0 to 60 cGy at three different dose rates: a low dose rate of 7.5 cGy/h, an intermediate dose rate of 30 cGy/h and a high dose rate of 70 cGy/min. The effect of 15 g/cm2 aluminum shielding on the induction of chromosome aberrations was investigated for each dose rate. After exposure, lymphocytes were incubated in growth medium containing phytohemagglutinin (PHA) and chromosome spreads were collected using a chemical induced premature chromosome condensation (PCC) technique. Aberrations were analyzed using the fluorescence in situ hybridization (FISH) technique with three different colored chromosome-painting probes. The frequency of reciprocal and complex-type chromosome exchanges were compared in shielded and unshielded samples. PMID- 12539754 TI - Monte Carlo transport model comparison with 1A GeV accelerated iron experiment: heavy-ion shielding evaluation of NASA space flight-crew foodstuff. AB - Deep-space manned flight as a reality depends on a viable solution to the radiation problem. Both acute and chronic radiation health threats are known to exist, with solar particle events as an example of the former and galactic cosmic rays (GCR) of the latter. In this experiment Iron ions of 1A GeV are used to simulate GCR and to determine the secondary radiation field created as the GCR like particles interact with a thick target. A NASA prepared food pantry locker was subjected to the iron beam and the secondary fluence recorded. A modified version of the Monte Carlo heavy ion transport code developed by Zeitlin at LBNL is compared with experimental fluence. The foodstuff is modeled as mixed nuts as defined by the 71st edition of the Chemical Rubber Company (CRC) Handbook of Physics and Chemistry. The results indicate a good agreement between the experimental data and the model. The agreement between model and experiment is determined using a linear fit to ordered pairs of data. The intercept is forced to zero. The slope fit is 0.825 and the R2 value is 0.429 over the resolved fluence region. The removal of an outlier, Z=14, gives values of 0.888 and 0.705 for slope and R2 respectively. PMID- 12539755 TI - Major Depressive Disorder (MDD) is associated with a primary defect within the serotonergic system. PMID- 12539756 TI - A 17-year-old patient with pulmonary infiltrate and rash. PMID- 12539757 TI - Nuclear model calculations and their role in space radiation research. AB - Proper assessments of spacecraft shielding requirements and concomitant estimates of risk to spacecraft crews from energetic space radiation requires accurate, quantitative methods of characterizing the compositional changes in these radiation fields as they pass through thick absorbers. These quantitative methods are also needed for characterizing accelerator beams used in space radiobiology studies. Because of the impracticality/impossibility of measuring these altered radiation fields inside critical internal body organs of biological test specimens and humans, computational methods rather than direct measurements must be used. Since composition changes in the fields arise from nuclear interaction processes (elastic, inelastic and breakup), knowledge of the appropriate cross sections and spectra must be available. Experiments alone cannot provide the necessary cross section and secondary particle (neutron and charged particle) spectral data because of the large number of nuclear species and wide range of energies involved in space radiation research. Hence, nuclear models are needed. In this paper current methods of predicting total and absorption cross sections and secondary particle (neutrons and ions) yields and spectra for space radiation protection analyses are reviewed. Model shortcomings are discussed and future needs presented. PMID- 12539758 TI - Calibration results obtained with Liulin-4 type dosimeters. AB - The Mobile Radiation Exposure Control System's (Liulin-4 type) main purpose is to monitor simultaneously the doses and fluxes at 4 independent places. It can also be used for personnel dosimetry. The system consists of 4 battery-operated 256 channel dosimeters-spectrometers. We describe results obtained during the calibrations of the spectrometers at the Cyclotron facilities of the University of Louvain, Belgium and of the National Institute of Radiological Sciences-STA, Chiba, Japan with protons of energies up to 70 MeV. The angular sensitivities of the devices are studied and compared with Monte-Carlo predictions. We also present the results obtained at the HIMAC accelerator with 500 MeV/u Fe ions and at the CERN high energy radiation reference fields. Records made during airplane flights are shown and compared with the predictions of the CARI-6 model. PMID- 12539759 TI - Solid-state integrating detectors as an indicator of biological doses from HZE particles. AB - For interpretation of results obtained in future biological experiments in the International Space Station (ISS), biologically equivalent doses have to be determined using small-scale detectors without disturbing the surrounding radiation field. The detectors should be lightweight, stable, safe, and simple in handling. Solid-state integrating detectors (SSID) can satisfy these requirements. This paper demonstrates that combination of SSID such as thermoluminescence dosimeters and radiophotoluminescence glasses can be practically used for the evaluation of biologically equivalent doses. Statistical errors (type-A uncertainty) of this method will be satisfactorily small relative to those generally observed in biological responses. Permissible levels of systematic errors (type-B uncertainty) depend on dosimetry purposes (most probable or conventional) and variability of biological responses. PMID- 12539760 TI - Bronchoscopy in a 12-month-old boy being treated for leukaemia. PMID- 12539761 TI - Parapneumonic pleural effusion. PMID- 12539762 TI - A new perspective of carcinogenesis from protracted high-LET radiation arises from the two-stage clonal expansion model. AB - When applied to the Colorado Plateau miner population, the two-stage clonal expansion (TSCE) model of radiation carcinogenesis predicts that radiation induced promotion dominates radiation-induced initiation. Thus, according to the model, at least for alpha-particle radiation from inhaled radon daughters, lung cancer induction over long periods of protracted irradiation appears to be dominated by radiation-induced modification of the proliferation kinetics of already-initiated cells rather than by direct radiation-induced initiation (i.e., mutation) of normal cells. We explore the possible consequences of this result for radiation exposures to space travelers on long missions. Still unknown is the LET dependence of this effect. Speculations of the cause of this phenomenon include the suggestion that modification of cell kinetics is caused by a "bystander" effect, i.e., the traversal of normal cells by alpha particles, followed by the signaling of these cells to nearby initiated cells which then modify their proliferation kinetics. PMID- 12539763 TI - Large pleural effusion. PMID- 12539764 TI - Langerhans cell histiocytosis. PMID- 12539765 TI - Radiation protection guidance for activities in low-Earth orbit. AB - Scientific Committee 75 (SC 75) of the National Council on Radiation Protection and Measurements (NCRP) was assembled for the purpose of providing guidance to NASA concerning radiation protection in low-Earth orbit. The report of SC 75 was published in December 2000 as NCRP Report No. 132. In this presentation an overview of the findings and recommendations of the committee report will be presented. PMID- 12539766 TI - A comparison of quality factors and weighting factors for characterizing astronaut radiation exposures. AB - Radiation exposures are typically characterized by two quantities. The first is the absorbed dose, or the energy deposited per unit mass for specific types of radiation passing through specified materials. The same amount of energy deposited in material by two different types of radiation, however, can result in two different levels of risk. Because of this, for the purpose of radiation protection operations, absorbed dose is modified by a second factor intended to normalize the risk associated with a given exposure. We present here an inter comparison of methods for this modification. First is the radiation quality factor (Q), as defined by ICRP publication 60. This quantity is related functionally to the unrestricted linear energy transfer (LET) of a given radiation, and is multiplied by the absorbed dose to derive the dose equivalent (H). The second method for modifying absorbed dose is the radiation weighting factor, also given in ICRP-60, or as modified in NCRP report 115. To implement the weighting factor, the absorbed dose resulting from incidence of a particular radiation is multiplied by a factor assigned to that type of radiation, giving the equivalent dose. We compare calculations done based on identical fields of radiation representative of that encountered by the MIR space station, applying each of these two methods. PMID- 12539767 TI - US orthodontist's reluctance to include autotransplantation. PMID- 12539768 TI - Buying staff not the solution. PMID- 12539769 TI - Different logic might provide new answers. PMID- 12539770 TI - The cervical vertebral maturation method: some need for clarification. PMID- 12539771 TI - Objectivity and worldly wisdom: missing ingredients in orthopaedic training and practice. PMID- 12539772 TI - Solar particle event organ doses and dose equivalents for interplanetary crews: variations due to body size. AB - Proper assessments of spacecraft shielding requirements and concomitant estimates of risk to critical body organs of spacecraft crews from energetic space radiation require accurate, quantitative methods of characterizing the compositional changes in these radiation fields as they pass through the spacecraft and overlying tissue. When estimating astronaut radiation organ doses and dose equivalents it is customary to use the Computerized Anatomical Man (CAM) model of human geometry to account for body self-shielding. Usually, the distribution for the 50th percentile man (175 cm height; 70 kg mass) is used. Most male members of the U.S. astronaut corps are taller and nearly all have heights that deviate from the 175 cm mean. In this work, estimates of critical organ doses and dose equivalents for interplanetary crews exposed to an event similar to the October 1989 solar particle event are presented for male body sizes that vary from the 5th to the 95th percentiles. Overall the results suggest that calculations of organ dose and dose equivalent may vary by as much as approximately 15% as body size is varied from the 5th to the 95th percentile in the population used to derive the CAM model data. PMID- 12539773 TI - The dependence of solar energetic particle fluxes in the Earth-Mars-Earth route on solar activity period. AB - This report presents the results of analyzing the relative importance of particle fluxes of different origin in the Earth-Mars-Earth route during different solar activity periods. The analysis has been made in terms of the galactic cosmic ray and solar energetic particle flux models developed at Moscow State University. The results demonstrate the extreme importance of the high-energy solar particle fluxes in interplanetary space even during the years of "quiet" Sun. PMID- 12539774 TI - Estimates of radiation effect for a spacecraft on the Earth-Mars-Earth route. AB - The space radiation environment predicted for a spacecraft on the Earth-Mars Earth route at different solar activity levels is analyzed in terms of the Russian-devised models. PMID- 12539775 TI - Radiation risk during long-term spaceflight. AB - Cosmonauts' exposure to cosmic rays during long-term spaceflight can cause unfavorable effects in health and risk for the crew members' lives. All unfavorable effects induced by exposure should be taken into consideration for the risk estimation. They should include both the acute deterministic effects and delayed effects called stochastic. On the ground the limitation of unfavorable consequences of acute exposure is achieved by means of establishing dose limits. But in space applications this approach can't be acceptable. Establishing a fixed dose limit is adequate to introducing indefinite reserve coefficient and therefore ineffective usage of spacecraft resource. The method of radiation risk calculation caused by acute and delayed effects of cosmonauts' exposure is discussed and substantiated in the report. Peculiarities of the impact of permanent radiation sources (galactic cosmic rays and trapped radiation) and the variable one (solar cosmic rays) are taken into consideration. PMID- 12539776 TI - Estimation of a cosmonaut's radiation hazard during long-term space missions on the basis of a generalized dosimetric function. AB - This paper presents a new concept of radiation hazard assessment for spacecraft crew members during long term space missions on the basis of a generalized dosimetric function. This new dosimetric function enables a complicated nature of space radiation exposure to be reduced to the conditions of a standard irradiation. It can be obtained on the basis of mean-tissue equivalent dose values calculated for each space radiation source and transmission coefficients describing the influence of the complex spatial and temporal distribution of the absorbed dose in the cosmonaut's body on the radiobiological effects. The combination of cosmic ionizing radiation with other non-radiation nature factors in flight can also be accounted for. In terms of the generalized dose, it is possible to assess the nature and extent of lowering a crew working capacity, as well as radiation risk, both during a flight and post flight period. PMID- 12539777 TI - Lifetime total radiation risk of cosmonauts for orbital and interplanetary flights. AB - This paper presents results of calculations of total radiation risk for cosmonauts over their lifetimes and assessments of possible shortening of life expectancy on the basis of generalized doses calculated for cosmonauts after a long term interplanetary and orbital space missions on "MIR" station and International Space Station with the use of mathematical expressions coming from a model of change mortality rate of mammals after irradiation. Tumor risk assessments for cosmonauts over lifetime after flights are also given. Dependences of the delayed radiation consequences mentioned above on flight duration, spacecraft shielding thicknesses, solar activity and cosmonauts' age are analyzed. PMID- 12539778 TI - Mathematical model for assessment of radiation risk on long space missions. AB - A mathematical model is developed which describes the dynamics of radiation induced mortality in mammalian populations. It relates statistical biometric functions with statistical characteristics and dynamics of an organism's critical system. In the framework of the model the effects of low and very low dose rates of chronic radiation on mice are simulated. Respectively, thrombocytopoietic and granulocytopoietic systems are considered as the critical ones. To calculate the dynamics of these systems, mathematical models are applied, too. In accordance with experimental data, the mortality model reproduces on quantitative level both increased and decreased mortality rates in populations of LAF1 mice, which were chronically exposed, respectively, to low and very low level radiation. All this makes it feasible to use the model as a basis for risk assessments of low level long-term irradiation. PMID- 12539779 TI - Probability of cell hits in selected organs and tissues by high-LET particles at the ISS orbit. AB - The fluence of high-LET particles (HLP) with LET infinity H2O greater than 15 keV micrometers-1 in selected organs and tissues were measured with plastic nuclear track detectors using a life-size human phantom on the 9th Shuttle-Mir Mission (STS-91). The planar-track fluence of HLP during the 9.8-day mission ranged from 1.9 x 10(3) n cm-2 (bladder) to 5.1 x 10(3) n cm-2 (brain) by a factor of 2.7. Based on these data, a probability of HLP hits to a matured cell of each organ or tissue was roughly estimated for a 90-day ISS mission. In the calculation, all cells were assumed to be spheres with a geometric cross-sectional area of 500 micrometers2 and the cell-hit frequency from isotropic space radiation can be described by the Poisson-distribution function. As results, the probability of one or more than 1 hit to a single cell by HLP for 90 days ranged from 17% to 38%; that of two or more than 2 hits was estimated to be 1.3-8.2%. PMID- 12539780 TI - New minor taxanes analogues from the needles of Taxus canadenis. AB - Seven new taxanes were isolated from the needles of the Canadian yew: unusual functional groups, positions and/or stereochemical features are described. Their chemical structures were rigorously characterized by detailed high resolution NMR analyses and confirmed by high resolution Fast Atom Bombardment Mass Spectrometry. Unlike paclitaxel and taxuspine D, these taxanes had no effect on tubulin assembly. PMID- 12539781 TI - Radiation-induced health effects on atmospheric flight crew members: clues for a radiation-related risk analysis. AB - There are few human data on low-dose-rate-radiation exposure and the consequent acute and late effects. This fact makes it difficult to assess health risks due to radiation in the space environment, especially for long-term missions. Epidemiological data on civilian flight personnel cohorts can provide information on effects due to the low-dose and low-dose rate mixed high- and low-LET radiation environment in the earth's atmosphere. The physical characteristics of the radiation environment of the atmosphere make the results of the studies of commercial flight personnel relevant to the studies of activities in space. The cooperative international effort now in progress to investigate dose reconstructions will contribute to our understanding of radiation risks for space exploration. PMID- 12539782 TI - Effects of gamma-ray and high energy carbon ion irradiation on swimming velocity of Euglena gracilis. AB - The effects of gamma-ray and high energy carbon ion irradiation on the swimming velocity of the photosynthetic flagellate Euglena gracilis strain Z were studied, focusing on a dose-effect relationship. Cells were exposed to 60Co gamma-rays at 6 doses of 10, 15, 20, 40, 100 and 200 Gy for water, and also to 290 MeV/amu carbon ions from the Heavy Ion Medical Accelerator in Chiba at 7 doses (5, 10, 15, 20, 50, 100 and 200 Gy for water). The swimming velocity was measured by a biomonitoring system, called ECOTOX. The swimming velocities of Euglena gracilis cells were significantly decreased by >40 Gy gamma-rays and >5 Gy carbon ions, respectively. The 50% effective doses for inhibition, 34 +/- 4 Gy (gamma-rays) and 13 +/- 1 Gy (290 MeV/amu carbon ions), were estimated from the best fit to data of the logistic model. The relative biological effectiveness (2.6 +/- 0.4) was calculated by the ratio of 50% effective doses. The inhibition of the swimming velocity of the cells irradiated with gamma-rays was still present after 3 days, while recovery of the swimming velocity was shown in the cells exposed to 290 MeV/amu carbon ions. It is suggested that ionizing radiation inhibits ATP production and/or increases frictional drag on beating of the flagellum, thus decreasing swimming velocity. PMID- 12539783 TI - [Band-like alopecia of the forehead-hair zone. Postmenopausal frontal fibrosing alopecia]. PMID- 12539784 TI - A review of solar proton events during the 22nd solar cycle. AB - Solar cycle 22 had significant, large fluence, energetic particle events on a scale reminiscent of the 19th solar cycle. Examination of the characteristics of these large events suggests that some of the old concepts of spectral form, intensity-time envelope and energy extrapolations, used to estimate the dose from large events that occurred during previous solar cycles should be re-evaluated. There has also been a dramatic change in perspective regarding the source of solar protons observed in interplanetary space. Very large fluence events are associated with powerful fast interplanetary shocks. The elemental composition and charge state of these events is suggestive of a dominate source in the solar corona and not from a very hot plasma. Furthermore, there is a strong suggestion that the intensity-time profile observed in space is dominated by the connection of the observer to an interplanetary shock source rather than to a unique location near the surface of the sun. These concepts will be examined from the perspective of energetic particles contributing to the dose experienced by an astronaut on an interplanetary space mission. PMID- 12539785 TI - The cosmic ray ground level enhancement of 6 November 1997. AB - The relativistic solar proton event of 6 November 1997 resulted in the first ground-level enhancement (GLE) of solar cycle 23. The earliest onset was around 1215 UT but was up to 15 minutes later at some neutron monitor locations. The time of maximum intensity also varied significantly over the world-wide neutron monitor network. The modeled particle distributions and spectra are presented. The apparent particle arrival direction is found to be largely consistent with propagation outward from the sun along interplanetary magnetic field lines. PMID- 12539786 TI - Re-analysis of the cosmic ray ground level enhancement of 4 May 1960. AB - The relativistic solar particle event of 4 May 1960, resulting in a cosmic ray ground level enhancement, occurred well before modern analysis techniques were available. We have located surviving data from 23 neutron monitors and have used these to estimate the spectrum, mean arrival direction and particle pitch angle distribution as the event progressed. We find that the apparent particle arrival direction was at equatorial latitudes, over northern South America, in contrast to contemporary analyses that proposed it to be over North America. Our modified power law spectra are broadly consistent with earlier results. Data from stations above sea level need to be corrected for altitude using a two-attenuation length technique. The standard method involves comparison of data from two relatively close stations at significantly different altitude. We have shown that this method may be unreliable in cases, such as this, of quite sharp anisotropy. PMID- 12539787 TI - Laser capture microdissection as an aid to ultrastructural analysis. AB - Laser capture microdissection uses a microscope to identify specific cells for microdissection and then a laser-sensitive plastic to capture and remove the cells from their substrate. This efficient capture method was originally developed to capture cells for genetic analysis. However, it has also been used to capture cells for proteonomic analysis. In this article, we extend the uses of laser-capture microdissection by reporting a method for preparing captured cells for ultrastructural analysis by transmission electron microscopy. Cells prepared by our methodology show good fine structure preservation and are easily sectioned by standard ultramicrotomy. PMID- 12539788 TI - The digital microscopy classroom. AB - This paper describes the implementation and functionality of a base-band network for the remote control of scientific instruments, in particular scanning and transmission electron microscopes. The base-band network is a hard-wired connection between keyboard and pointing devices from the remote computers to the CPU of the instrument computer; the video signal is then routed back to the remote user and to a high resolution projection system. The system is independent of software and hardware platform, and is highly reliable. PMID- 12539789 TI - The influence of daily hassles and role balance on health status: a study of mothers of grade school children. AB - The purpose of this study was to investigate the influence of daily hassles and role balance on health status. Participants (146 mothers) completed a self administered survey that included assessments of daily hassles, role balance, and health status. Based on hierarchal regression analyses with interaction terms, daily hassles and role balance directly influenced the mothers' health status. The results also indicated that role balance had a buffering effect on the relationship between daily hassles and health status. These findings lend empirical support to the theory of role balance. PMID- 12539790 TI - Forces promoting health insurance coverage of homebirth: a case study in Washington State. AB - The movement of childbirth to the hospital in the early 1900s and the eventual establishments of health insurance reimbursement for hospital birth--but not for homebirth--solidified and reflected physician dominance in the area of obstetrics. Until recently, it was rare that a health insurer or a health maintenance organization (HMO) would cover a homebirth. However, in Washington State the majority of health insurance groups cover homebirths, which are generally attended by licensed midwives. In this context, our research is a case study focused on answering the question: What are the forces promoting the extensive coverage of homebirth by health insurers in Washington State? Data were gathered primarily through fourteen (14) in-depth, audiotaped interviews with key informants in relevant agencies and organizations in the state (i.e., state offices; midwife and other professional associations; and health insurance groups). Results suggest that consumer demand was an important precipitating factor without which changes to health insurance coverage would likely not have been made. Additionally, changes in state policies and professional mobilization on the part of licensed midwives were critical factors facilitating the widespread reimbursement for homebirth. Health care organizations' concerns for cost containment had little impact on this health insurance trend. Our study concludes that jurisdictional openings in the system of professions can be facilitated by a small number of strategically positioned individuals. PMID- 12539791 TI - Power, empowerment, and critical consciousness in community collaboration: lessons from an advisory panel for an HIV awareness media campaign for women. AB - This study examines the interplay of power, empowerment, and critical consciousness in the dynamics of a public health community collaboration around an HIV prevention media campaign for women. Methodology included a process evaluation consisting of participant observation, focus groups, and one-on-one interviews with advisory board members. Findings suggest that attention to the dynamics of power should begin at the very earliest levels of planning collaborative health projects for women. Additionally, findings indicate that the invisibility of power and privilege affect women's participation in collaborative projects. It is recommended that intentional educational efforts be implemented to help participants in collaborative health projects for women develop the necessary skills and awareness to create an atmosphere of respect and mutuality rather than one of domination and subordination. Education for critical consciousness is suggested as one means for achieving this goal. PMID- 12539792 TI - Double trouble: violent and non-violent traumas among women at sexual risk of HIV infection. AB - BACKGROUND: This study examines the association between trauma and HIV risk behaviors among women at sexual risk for HIV infection. METHODS: From April to August 1998, high-risk HIV negative women were recruited in the South Bronx into a year-long cohort study. At the 12-month visit, 116 women were interviewed face to-face about recent and lifetime violent and non-violent traumas. RESULTS: The women reported a substantial prevalence of sexual risk behaviors associated with the acquisition of HIV. At baseline, almost two-thirds (64%) reported unprotected vaginal sex in the previous six months, and in the previous year, 62% had smoked crack, 52% reported sex-for-money-or-drugs exchanges, and 47% had five or more male sex partners. The lifetime prevalence of trauma was high: 81% had experienced one or more violent traumas and 97% had experienced one or more non violent traumas. Women who had experienced violent trauma--physical assault by a partner (OR = 2.88; 95% CI 1.12; 7.41)--and those who had experienced non-violent trauma--loss of a child to foster care (OR = 3.34; 95% CI 1.04; 10.65)--were more likely to use crack than others. Those who had experienced non-violent trauma, by witnessing a physical assault (OR = 2.31; 95% CI 0.99; 5.40), were also more likely than others to have exchanged sex. CONCLUSIONS: Both violent and non violent traumas appear to play a role in the behaviors that place women at risk of HIV infection, particularly using crack and exchanging sex. PMID- 12539793 TI - Perceptions of risk factors for breast cancer and attitudes toward mammography among women who are current, ex- and non-smokers. AB - Understanding risk perceptions for breast cancer among women smokers is important because smokers tend to underutilize breast cancer screening. Perceptions of the relative importance of a variety of factors which may increase breast cancer risk and the benefits/barriers of mammography, were examined among women who were current (n = 185), ex- (n = 632) and never (n = 623) smokers. Participants were a subset of women taking part in a project to increase mammography utilization among women aged 50 and over. Current smokers, but not ex-smokers, were significantly less likely than never smokers to agree that health risk behaviors most frequently seen in smokers (e.g., smoking cigarettes, high-fat diet, low intake of fruits and vegetables, physical inactivity, drinking alcohol) may increase risk for breast cancer. Current smokers, but not ex-smokers, perceived more barriers and fewer benefits of mammography, than never smokers. PMID- 12539794 TI - Worry regarding major diseases among older African-American, Native-American, and Caucasian women. AB - This study examined worry regarding seven major diseases and their correlates in a sample of African-American (n = 57), Native-American (n = 50), and Caucasian (n = 53) women ages 36 to 91 years. African-American and Native-American women were most worried about developing cancer (44% and 50%, respectively) while Caucasian women were most worried about osteoporosis (37%) and cancer (33%). Women from each ethnic group were more worried about developing cancer than cardiovascular diseases and conditions. African-American and Native-American women were more worried than Caucasian women about developing diabetes and high cholesterol. Body mass index (BMI) was a consistent correlate of worry: heavier women were more worried about developing diseases than were leaner women. Other risk factors (e.g., physical activity, blood pressure), however, were generally not associated with disease worry. In fact, age was inversely associated with worry regarding diabetes, cancer, and osteoporosis. Although women who were more worried about developing cancer were more likely to perform monthly breast self-exams, worry regarding other diseases was not associated with preventive actions. These results are generally consistent with other studies that indicate women are more concerned about cancer than cardiovascular diseases. PMID- 12539795 TI - Relationships between body image and depressive symptoms during postpartum in ethnically diverse, low income women. AB - The aim of this study was to examine body image, depressive symptoms, and their interrelationship at post-delivery and at 6 weeks postpartum in a triethnic sample of low income new mothers. Data for this study were drawn from the Austin New Mothers Study. Participants were 76 Anglo/White, 72 African American, and 135 Hispanic new mothers who were participating in Medicaid, at least 18 years old, free of medical risk factors, and delivered at term. Body image was measured by the Body Cathexis Scale (BCS) and depressive symptoms by the Center for Epidemiologic Studies Depression Scale (CES-D). The leading areas of body dissatisfaction in postpartum were similar across ethnic groups. At 6 weeks postpartum Anglo women had the highest number of body image components perceived negatively, whereas African-American women had the least. In the overall sample, body image attitudes were significantly related to depressive symptoms in correlational (r's .19 to .34) and regression analyses. In addition, African American ethnicity was associated in regression analyses with higher depressive symptoms. Neither African American nor Hispanic ethnicity significantly moderated the relationship between body image attitudes and depressive symptoms in tests of interactions. Marital/partnered status and income level were related to depressive symptoms at both time points. In addition, being a woman with a parity of II was associated with higher depressive symptoms at 6 weeks postpartum. Although addition of these personal variables increased the percent of variance accounted for, body image attitudes continued to be significant predictors of depressive symptoms. PMID- 12539796 TI - Improving exposure assessment requires measurements and modeling. PMID- 12539797 TI - Dermal exposure assessment in occupational epidemiologic research. AB - Recognition of the importance of skin exposure in industrial settings has steadily increased over the last few decades. Unfortunately, the growing attention to dermal exposure in industrial hygiene has often not been reflected in the field of occupational epidemiology. An extensive literature survey was conducted to identify dermal exposure assessment methods that have been applied in epidemiologic studies. Subsequently, methodologies are postulated that could be applied to epidemiologic research. Attention is given to intensity, frequency, and duration of exposure, the exposed surface area, and personal, temporal and spatial variability in dermal exposure and uptake. It is anticipated that, in the near future, dermal exposure assessment in epidemiologic research will be based generally on expert judgment and to some degree on process-specific exposure models. Field studies collecting quantitative dermal exposure data and statistical modeling to identify exposure determinants will, however, be imperative if progress is to be made in the field of dermal exposure assessment for epidemiologic purposes. PMID- 12539798 TI - Time control, catecholamines and back pain among young nurses. AB - OBJECTIVES: This study had two objectives. First, it addressed concern with the contribution of work stressors and resources to the development of back pain, over and above the influence of biomechanical work factors. Second, using recent models about the role of the sympathetic-adrenal medullar system in musculoskeletal problems as its basis, it tested whether low-back pain is associated with higher levels of catecholamines. METHODS: Altogether 114 nurses filled out a questionnaire in their first year of practice and again one year later. In addition, in a subsample of 24 nurses studied intensively at follow-up, urinary catecholamines were assessed at noon, before the end of work, in the evening, and at corresponding times on a day off. Daily stressful experiences and daily mood were also recorded. RESULTS: With control for baseline pain, biomechanical workload, and other potentially confounding variables, time control at the beginning of the study predicted low-back pain a year later. In the subsample, the epinephrine and norepinephrine levels were higher in those reporting more frequent episodes of back pain, the largest differences occurring at the end of work. In addition, control over stressful events at work was lower in this group. CONCLUSIONS: Time control is a risk factor for low-back pain among nurses beyond the influence of physical work load. Low control at work may increase the activity of the sympathetic-adrenal medullar system, which seems to play an important role in the development of musculoskeletal pain. PMID- 12539799 TI - Work-schedule characteristics and reported musculoskeletal disorders of registered nurses. AB - OBJECTIVES: The relationship between a combination of demanding work-schedule characteristics and reported musculoskeletal disorders of the neck, shoulders, and back was examined. METHODS: A probability sample of 1163 nurses, randomly selected from the list of actively licensed nurses in two states of the United States, served as the sample for this cross-sectional study. Data were collected via an anonymous survey mailed to the participants' homes from October 1999 through February 2000. RESULTS: Four of the nine work-schedule characteristics (working full-time, >8 hours/day, 2-4 weekends/month, and other than day shift) were significantly related to musculoskeletal disorders in one or more body sites. When a work-schedule index was created by summing the nine characteristics, a demanding schedule was significantly associated with musculoskeletal disorders in the neck [odds ratio (OR) 1.10, 95% confidence interval (95% CI) 1.00-1.21], shoulder (OR 1.12, 95% CI 1.01-1.23), and back (OR 1.16, 95% CI 1.06-1.27). Adjustment for psychological and physical job demands reduced the odds ratios slightly and therefore suggested that some of the association between musculoskeletal disorders and schedule was due to increased exposure to these job demands. Working "long hours" (>12 hours/day, >40 hours/week) and "off hours" (weekends and "other than day shifts") were associated with a 50-170% increase in the age-adjusted odds ratio for musculoskeletal disorders in the three body sites. CONCLUSIONS: The findings of this study suggest that preventing musculoskeletal disorders requires system level approaches to scheduling that reduce the time of exposure to demanding work conditions and promote healthful work-rest patterns. PMID- 12539800 TI - Exposure to carcinogenic air pollutants among policemen working close to traffic in an urban area. AB - OBJECTIVE: This study evaluates individual airborne exposure to gaseous and particulate carcinogenic pollutants in a group of policemen working close to traffic in the center of Grenoble, France. METHODS: Sixty-two personal active air samples were collected during the workshifts of eight policemen in summer and in winter during the occurrence of the thermal inversion phenomenon. Seventeen stationary air samples were monitored in the policemen's work area during the same period with the same sampling devices as used for the personal samples. The respirable particle concentration was determined using the gravimetric method. The concentrations of polycyclic aromatic hydrocarbons and aldehydes were measured with high-performance liquid chromatography with fluorimetric or ultraviolet detection. The benzene-toluene-xylene levels were determined with gas chromatography with flame ionization detection. RESULTS: The median concentration of the personal samples for respirable particles was 55.5 microg/m3 in the summer and 124 microg/m3 in the winter. The corresponding values were 0.10 and 0.28 ng/m3 for benzo(a)pyrene, 14 and 21 microg/m3 for formaldehyde, and 10.5 and 23.5 microg/m3 for benzene. The median personal concentrations were equal to or higher than the corresponding stationary levels for both seasons. Compared with the concentration of aldehydes, the concentrations of respirable particles, polycyclic aromatic hydrocarbons, and benzene-toluene-xylene appeared to show higher individual variability. CONCLUSIONS: The occupational exposure of policemen does not exceed any currently applicable occupational or medical exposure limits. Individual particulate levels should preferably be monitored in Grenoble in winter to avoid underestimations. PMID- 12539801 TI - Cancer incidence among workers in the asbestos-cement producing industry in Norway. AB - OBJECTIVES: The incidence of cancer among employees of a Norwegian asbestos cement factory was studied in relation to duration of exposure and time since first exposure. The factory was active in 1942-1968. Most of the asbestos in use was chrysotile, but for technical reasons 8% amphiboles was added. METHODS: For the identification of cancer cases, a cohort of 541 male workers was linked to the Cancer Registry of Norway. The analysis was based on the comparison between the observed and expected number of cancer cases. Standardized incidence ratios (SIR) and 95% confidence intervals (95% CI) were estimated. Period of first employment, duration of employment, and time since first employment were used as indicators of exposure. Poisson regression analysis was used for the internal comparisons. RESULTS: The standardized incidence ratio was 52.5 (95% CI 31.1 83.0) for pleural mesothelioma, on the basis of 18 cases. The highest standardized incidence ratio was found for workers first employed in the earliest production period (SIR 99.0, 95% CI 51.3-173). No peritoneal mesothelioma was found. The standardized incidence ratio for lung cancer was 3.1 (95% CI 2.14.3), but no dose-response effect was observed. The ratio of mesothelioma to lung cancer cases was 1:2. CONCLUSIONS: This study showed a high incidence of mesothelioma and a high ratio of mesothelioma to lung cancer among asbestos cement workers. The high incidence of mesothelioma was probably due to the fact that a relatively high proportion of amphiboles was used in the production process. PMID- 12539802 TI - Time to pregnancy among partners of men exposed to di(2-ethylhexyl)phthalate. AB - OBJECTIVES: This study assessed paternal occupational exposure to di(2 ethylhexyl)phthalate (DEHP) in association with reduced fertility. METHODS: Men working in three plants with DEHP exposure were studied retrospectively. Male and female employees and their partners were invited to participate if they had reported a pregnancy or an attempt to achieve a pregnancy. Postal questionnaires and telephone interviews were used to collect additional data from the men and women, respectively. Information on time to pregnancy was eligible for 326 pregnancies fathered by 193 men. Male exposure to DEHP during every month of their time to pregnancy was classified into one of three exposure categories. The exposure ranged from <0.1 to 2.1 mg/m3. The fathers of only four pregnancies had DEHP exposure of >0.5 mg/m3 during the time to pregnancy. The pregnancies of employed women with unexposed partners or pregnancies of employed men unexposed during the time to pregnancy formed the reference group. RESULTS: The fecundability ratio for time to pregnancy was 1.07 [95% confidence interval (95% CI) 0.84-1.35] for those with low exposure and 0.97 (95% CI 0.70-1.33) for the highly exposed after adjustment for the father's age, mother's age, and length of recall. When the analyses were restricted to first pregnancy, the fecundability ratio was 1.13 (95% Cl 0.83-1.56) for low exposure and 1.02 (95% CI 0.66-1.59) for high exposure. CONCLUSIONS: Time to pregnancy is not prolonged among couples with paternal exposure to DEHP at a mean exposure level of <0.5 mg/m3. PMID- 12539803 TI - Trends in inhalation exposure to hydrocarbons among commercial painters in The Netherlands. AB - OBJECTIVES: An attempt was made to develop a database for measurements of exposure to solvents that could be used as a tool in the historical exposure assessment of commercial painters participating in a health surveillance program. METHODS: The measurement data on personal exposure from six studies still available for Dutch commercial painters were collected into a database. The database was analyzed to identify time trends for the inhalation exposure levels of hydrocarbons and the production conditions that influence exposure levels among commercial painters in The Netherlands. RESULTS: Altogether 304 measurements of solvent exposure were collected between 1980 and 1999, providing data for 137 workers. Toluene was selected as a marker for solvent exposure, since hydrocarbon exposures appeared to be strongly correlated. Exposure to toluene measured during the application of solvent-based paints has declined by 12% per year. The use of solvent-based paints, painting in small rooms, house (versus shipyard) painting, and spray-painting were associated with increased exposures. Water-based paint was also associated with increased exposure to toluene, relative to tasks in which no paint was used. The exposure model for toluene explained 86% of the between-worker variance. In a subset of the data, we observed that a single cell model did not adequately describe total solvent exposure among painters, because of the stronger-than-expected positive effect of source strength and the lack of the protective effect of general ventilation. CONCLUSIONS: An exposure model was developed that can be used to predict the intensity of inhalation exposure to aromatic solvents among commercial painters in The Netherlands. PMID- 12539804 TI - Silicosis and end-stage renal disease. AB - OBJECTIVES: The objective of this study was to determine the incidence of renal disease among workers with silicosis. METHODS: A population of 1,328 workers with definite silicosis and adequate work history information, drawn from three states with silicosis surveillance systems, was followed. Renal disease was ascertained via linkage of the cohort with a United States register (which has existed since 1977) of end-stage renal disease. RESULTS: In the first analysis, it was assumed that the risk of end-stage renal disease began upon exposure to silica. In this analysis 12 cases of end-stage renal disease were found versus 15.6 expected, for a rate ratio of 0.77. Four cases of glomerular end-stage renal disease were found (standardized incidence ratio 2.65, 95% confidence interval 0.56-5.25). It is possible that some persons with end-stage renal disease died before being entered into the silicosis registers. In a second analysis, person-time at risk was assumed to begin at the date of entry into the silicosis register. A rate ratio of 1.67 (95% confidence interval 0.76-3.17) was found for end-stage renal disease on the basis of nine observed cases. CONCLUSIONS: The results do not clearly show that patients with silicosis have an excess of end-state renal disease, although they do suggest an excess of glomerular end-stage renal disease. Analyses were limited by small numbers and possible selection biases. PMID- 12539805 TI - Restricted weekend services result in delays in discharges from hospital. PMID- 12539806 TI - Therapy to reduce risk of coronary heart disease. AB - Recent primary and secondary intervention studies have shown that reduction of low-density lipoprotein cholesterol (LDL-C) with statins significantly reduced coronary heart disease (CHD) morbidity and mortality. However, many patients with dyslipidemia who have or are at risk for CHD do not reach target LDL-C goals. The recently updated National Cholesterol Education Program (NCEP) Adult Treatment Panel (ATP) III guidelines identify a group of patients at very high risk for CHD for more aggressive LDL-C reduction and reaffirm the importance of high-density lipoprotein cholesterol (HDL-C) by raising the categorical threshold to 40 mg/dl. Lipid-lowering therapy needs to be more aggressive in both primary and secondary prevention settings, and therapy should be considered to increase HDL-C as well as lower LDL-C in order to improve patient outcomes. Both combination therapy and the next generation of statins may provide improved efficacy across the dyslipidemia spectrum. PMID- 12539807 TI - Treatment satisfaction of internal versus external cardioversion in patients with chronic atrial fibrillation--a randomized, prospective, 28-day follow-up study. AB - BACKGROUND AND HYPOTHESIS: The use of internal cardioversion (IC) in chronic atrial fibrillation (AF) may be limited by procedure-related distress. This procedure may be efficacious but is not necessarily perceived as satisfactory by the patient because of the frequent procedure-related distress. METHODS: We compared treatment satisfaction in 55 patients with chronic AF referred for cardioversion (CV). Satisfaction with conventional external cardioversion (EC, n = 27) and low-energy IC (n = 28) was compared immediately after the approach and at 28-day follow-up. RESULTS: Four hours after CV, satisfaction scores (ranging from 0 to 7) were higher with EC (mean 6.1 standard deviation [SD] +/- 1.4) versus IC (m = 5.4 SD +/- 1.8) (p = 0.09). At 28 days, treatment satisfaction decreased in EC (m = 5.5 SD +/- 2.1) and increased in the IC group (5.7 SD +/- 1.8) because of a decrease in the trustful attitude subscale (p = 0.026) followed by tolerance for distressing factors (p = 0.059). The analysis of variance for repeated measures revealed a significant time by method interaction effect (p = 0.04). Patients prone to developing low treatment satisfaction (LTS) at follow-up were more anxious (p = 0.007) before treatment. They suffered more from sleeping disorders (p = 0.009) and considered their disease condition to be worse than that in their counterparts (p = 0.027). Low treatment satisfaction at 28 days was associated with anxiety (p = 0.017), depression (p = 0.01), and the perception of heart-related symptoms (p = 0.001). Multivariate analysis revealed the failure to maintain sinus rhythm (p = 0.001) as the most powerful contributor to LTS. CONCLUSIONS: The novel IC approach causes acute procedure-related distress but has no enduring negative psychological side effects. Despite a greater patient perception of distressing procedure-related factors during IC compared with EC, the IC approach provides a greater 28-day benefit in terms of tolerability and acceptance on the part of the patients. Failure to maintain sinus rhythm rather than the method applied contributes most to LTS. PMID- 12539808 TI - Effects of simvastatin, an HMG-CoA reductase inhibitor, in patients with hypertriglyceridemia. AB - BACKGROUND: Patients with elevated levels of serum triglycerides (TG) often have other associated lipid abnormalities (e.g., low levels of high-density lipoprotein cholesterol [HDL-C]) and are at increased risk of developing coronary heart disease. Although the therapeutic benefits of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) in hypercholesterolemic patients have been well established, less is known about the effects of statins in patient populations with hypertriglyceridemia. HYPOTHESIS: The purpose of this study was to evaluate the lipoprotein-altering efficacy of simvastatin in hypertriglyceridemic patients. METHODS: This was a multicenter, randomized, double-blind, placebo-controlled study. In all, 195 patients with fasting serum triglyceride levels between 300 and 900 mg/dl received once daily doses of placebo or simvastatin 20, 40, or 80 mg for 6 weeks. RESULTS: Compared with placebo, simvastatin treatment across all doses resulted in significant reductions (p < 0.05 - < 0.001) in serum levels of triglycerides (-20 to -31% decrease) and TG-rich lipoprotein particles. Significant (p < 0.001) reductions were also seen in low-density lipoprotein cholesterol (-25 to -35%) and non-HDL-C (-26 to -40%). Levels of HDL-C were increased (7-11%) in the simvastatin groups compared with placebo (p < 0.05 - < 0.001). CONCLUSION: The results of this study demonstrate the beneficial effects of simvastatin in patients with hypertriglyceridemia. PMID- 12539809 TI - Prevalence and risk factors for erectile dysfunction in men with diabetes, hypertension, or both diseases: a community survey among 1,412 Israeli men. AB - BACKGROUND: Erectile dysfunction (ED) and cardiovascular disease share common risk factors and may be further aggravated by medical treatment for reducing them. HYPOTHESIS: The study was undertaken to assess the prevalence of ED in patients with diabetes (DM), hypertension (HTN), or both diseases, and to evaluate the effect of patient age, medical treatment, and disease duration and control on the prevalence of ED in this population. METHODS: A group of 150 primary practitioners who had patients with known DM and/or HTN conducted a survey, utilizing IIEF-15, a 15-item multidimensional, self-administered questionnaire used for the clinical assessment of ED. RESULTS: In all, 1,412 patients were included: 37% had DM, 38% had HTN, and 25% had both diseases. Their mean age was 55, 58, and 60 years, and 62, 46, and 67% had some degree of ED, respectively. The prevalence of ED increased with age and disease duration in each age group and was higher in subjects with DM than in those with HTN, especially in those aged < 65 years. Poor glycemic control was associated with a higher prevalence rate of ED early in the course of the disease. There was no significant difference in the prevalence of ED according to type and number of antihypertensive drugs. CONCLUSIONS: Erectile dysfunction is common among patients at high risk for cardiovascular disease because of diabetes and/or HTN. Diabetic men are affected earlier than those with HTN. Given the high frequency of ED in young patients with these risk factors, physicians should encourage an open discussion on the subject during routine visits to promote early detection and treatment. PMID- 12539810 TI - Oxidized low-density lipoprotein and high-density lipoprotein cholesterol modulate coronary arterial remodeling: an intravascular ultrasound study. AB - BACKGROUND: Oxidized low-density lipoprotein (oxLDL) not only plays an important role in plaque formation, but also impairs the endothelium-dependent relaxation. Constrictive remodeling rather than intimal hyperplasia mainly contributes to restenosis after balloon angioplasty. Probucol (powerful antioxidant) reduced restenosis rate by improving constrictive remodeling. Thus, oxLDL may modulate coronary arterial remodeling. HYPOTHESIS: The study was designed for using intravascular ultrasound to test the hypothesis that arterial constrictive remodeling (CR) was associated with oxLDL in patients with coronary artery disease. METHODS: Intravascular ultrasound was performed in 36 patients with de novo atherosclerotic coronary. Remodeling was defined and evaluated as follows: remodeling index (RI) = lesion vessel area (VA)/(proximal reference VA + distal reference VA)/2. Constrictive remodeling (CR) was defined as remodeling index (RI) < 0.9. Neutral and expansive remodeling (NER) was defined as RI > or = 0.9. The level of plasma ox-LDL was measured by sandwich ELISA using the monoclonal antibody (DLH3)-recognized oxidatively modified lipoproteins and the antihuman apoprotein B monoclonal antibody. RESULTS: Neutral and expansive remodeling was found in 24 lesions, and CR in 12 lesions. Remodeling index was significantly lower in the CR group than in the NER group (0.8 +/- 0.1 vs. 1.0 +/- 0.1, p < 0.001). The level of oxLDL in the CR group was significantly higher than that in the NER group (24.0 +/- 12.1 vs. 16.4 +/- 6.2 U/ml, p < 0.05). The level of high density lipoprotein-cholesterol (HDL-C) in the CR group was significantly lower than that in the NER group (40.5 +/- 4.8 vs. 46.2 +/- 10.6 mg/ml, p < 0.05). There was a statistically significant correlation between the value of HDL-C/ ox LDL and the RI (r = -0.48, p < 0.005). CONCLUSIONS: Oxidized LDL and HDL-C were associated with arterial remodeling in de novo atherosclerotic lesions. PMID- 12539811 TI - Electrocardiographic and vectorcardiographic findings of patients undergoing reductive ventriculectomy (Batista operation). AB - BACKGROUND: The Batista operation, or partial ventriculectomy, has been designed by the Brazilian surgeon Dr. Randas Batista as an alternative method for treating patients with idiopathic dilated cardiomyopathy. HYPOTHESIS: This study aimed to analyze electro- and vectorcardiographic data obtained from patients who underwent such surgery, and to evaluate its electrical repercussions on the heart. METHODS: Pre- and postoperative electrocardiography (ECG) and vectorcardiography (VCG) were performed 45 days apart in 15 patients undergoing reductive ventriculectomy. RESULTS: All ECGs showed sinus rhythm, with unchanged QRS duration. Left atrial enlargement (86.6%) and left ventricular hypertrophy (100%) were the most common findings. Bundle-branch blocks (BBBs) were often demonstrated on preoperative studies, predominantly (46.7%) left BBBs. Left anterior fascicular blocks were seen in four patients (26.6%), one associated with right BBB. Electrocardiographic changes suggestive of myocardial infarction (MI) were seen in four patients preoperatively; postoperatively, all had extended to or within the lateral wall. Five additional patients developed lateral MIs postoperatively, for a total of 9 patients (60%) with postoperative signs of infarction (p < 0.05). Electro- and vectorcardiography also showed significant postoperative lowering of QRS voltages (mean 40.74%) in all patients and a consequent difficulty to diagnose left ventricular hypertrophy, although the morphologic features did not change. CONCLUSIONS: These important ECG and VCG alterations are reflections of both the surgical technique and its clinical consequences. PMID- 12539812 TI - Images in cardiology: mycotic aneurysm and arteriocaval fistula. PMID- 12539813 TI - Isolated noncompaction left ventricular myocardium and polymorphic ventricular tachycardia. AB - A 57-year-old woman with syncope was admitted. She had a family history of sudden death: two brothers had died suddenly at the age of 47. Transesophageal echocardiography showed numerous prominent trabeculations and deep intertrabecular recesses in the anterior and lateroapical zones. Isotopic left ventricular ejection fraction was 46%. Cardiac catheterization showed coronary arteries with no angiographic lesions. A prominent trabecular zone and deep intertrabecular recesses were seen in the anterior wall on left ventriculography. Right ventriculography was normal. The diagnosis of isolated noncompaction left ventricular myocardium was established. Continuous 24-h electrocardiographic registry showed episodes of polymorphic ventricular tachycardia. Programmed ventricular stimulation performed at the right ventricular apex with up to three extrastimuli failed to induce ventricular arrhythmias. Treatment with beta blockers was initiated, but short runs of polymorphic ventricular tachycardia persisted. A dual-chamber automatic implantable defibrillator was implanted. We discuss the physiopathology of the arrhythmia. It appears that several factors could be responsible for the malignant arrhythmias in this entity. PMID- 12539814 TI - Profiles in cardiology. Rene Descartes. PMID- 12539815 TI - Understanding risk in hypercholesterolemia. AB - Atherosclerosis was relatively uncommon 100 years ago, when researchers first established its link to elevated cholesterol. As the twentieth century progressed, however, factors such as high-fat diets, sedentary lifestyles, cigarette smoking, and urbanization combined to increase the prevalence of both hypercholesterolemia and coronary heart disease (CHD) throughout the developed world. Atherogenesis begins at an early age and progresses throughout life, and cholesterol levels during young adulthood strongly predict the risk of CHD and related mortality during the ensuing decades. The total cholesterol level in youth also determines the actual age at which a critical level of atherosclerosis will be reached. Early studies on the primary and secondary prevention of CHD failed to identify a linear relationship between lipid lowering and risk reduction, primarily because older lipid-lowering agents lacked the potency to reduce cholesterol levels significantly enough to achieve lower cardiovascular event and mortality rates. The introduction of the statins, with their powerful lipid-lowering activity, overcame this limitation. Several large-scale trials of statins firmly established the efficacy of these agents in both primary and secondary CHD prevention. With the availability of statin therapy, we are now able to reduce the risk of major adverse CHD events by an average of 30%, regardless of patient age or gender. PMID- 12539816 TI - New dimension of statin action on ApoB atherogenicity. AB - Newer, more effective statins are powerful agents for reducing elevated levels of low-density lipoprotein (LDL) cholesterol and thereby lowering the risk of coronary heart disease (CHD) and related adverse events. Although LDL remains the primary target of therapy for reducing CHD risk, increased interest is focusing on apolipoprotein B (apoB)-containing lipoprotein subfractions--particularly very low-density lipoprotein (VLDL). VLDL remnants, and intermediate-density lipoproteins (IDL)--as secondary targets of therapy. Elevated apoB is known to be an important risk factor for CHD, and dysregulation of the metabolism of apoB containing lipoproteins is involved in the progression of atherosclerosis. Statins reduce circulating concentrations of atherogenic apoB-containing lipoproteins by decreasing the production of VLDL in the liver and, thus, the production of VLDL remnants and LDL. Statins also increase the clearance of these particles through upregulation of LDL receptors in the liver. Efforts to develop statins with enhanced lipid-modifying properties are ongoing. The optimal statin would offer a high degree of inhibition of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, a prolonged duration of action, hepatic selectivity for maximal upregulation of LDL receptors, and a low potential for drug-drug interactions. Recent studies have shown that rosuvastatin, a new agent in this class, demonstrates these qualities. Rosuvastatin is a highly effective inhibitor of HMG-CoA reductase, is relatively nonlipophilic, has a half-life of approximately 20 h, exhibits hepatic selectivity, has little systemic availability, and has a low potential for drug-drug interactions because of its limited degree of metabolism by the cytochrome P450 system. A recent double blind, crossover study revealed that treatment with rosuvastatin resulted in marked reductions in apoB-containing lipoproteins in patients with type IIa or IIb dyslipidemia. By reducing the number of atherogenic lipoprotein particles, rosuvastatin decreases the atherosclerotic burden in hyperlipidemic patients at high risk for CHD and related adverse outcomes. PMID- 12539817 TI - Effects of statins in reducing thrombotic risk and modulating plaque vulnerability. AB - A large body of evidence demonstrates that statin therapy reduces risk for coronary events. This benefit probably stems in large part from lipid lowering, but lipid-independent cellular effects may also contribute. Statin therapy may lower the risk of thrombosis by reducing tissue factor expression and increasing plasminogen activators while reducing plasminogen-activator inhibitor. The statins may also improve endothelial function and reduce inflammatory response by increasing nitric oxide activity; all statins tested can decrease levels of C reactive protein, a systemic marker of inflammation. Indeed, limiting inflammation may prove an important mechanism of statins' clinical benefits due to both lipid lowering and direct cellular effects. For example, attenuation of inflammation probably promotes maintained integrity of the fibrous cap of the atherosclerotic lesion by inhibiting processes that degrade the collagenous structure of the cap. Rupture of the fibrous cap causes most fatal coronary thrombosis. PMID- 12539818 TI - Comparing HMG-CoA reductase inhibitors. AB - The statins have proved to be some of the most potent therapies for reducing elevated low-density lipoprotein (LDL) cholesterol and lessening the risk of coronary heart disease (CHD) and related events. Nonetheless, there are still questions about the clinical relevance of individual drug characteristics, such as chemical derivation, solubility properties, and metabolic route, in terms of tolerability or therapeutic benefit. At the same time, no clear explanation has emerged for the significantly steeper reductions in LDL cholesterol levels achieved with atorvastatin versus lovastatin, simvastatin, pravastatin, or fluvastatin, or, more recently, with rosuvastatin versus atorvastatin, although possible mechanisms have been suggested. More studies are needed to characterize the effects of statins on high-density lipoprotein (HDL) in different patient groups. Clearly, though, several statins have yielded significant reductions in CHD risk and have shown to be well tolerated in both primary and secondary prevention trials. The possibility that statins exert pronounced effects beyond lowering blood lipids is opening other avenues of research into the benefits of these drugs. PMID- 12539819 TI - Treating hypercholesterolemia: looking forward. AB - Despite important advances in the management of hypercholesterolemia in recent decades, many patients with lipid disorders remain unidentified or undertreated and so continue to have unfavorable levels of low-density lipoprotein (LDL) cholesterol and an increased risk for coronary events. The statins--which inhibit 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the rate-limiting enzyme in cholesterol biosynthesis--have proved to be the most powerful pharmacologic agents for lowering serum lipids, and newer statins offer even greater efficacy than the agents introduced 10 to 15 years ago. Studies have shown that rosuvastatin, in late-stage development, is a very potent agent for the treatment of primary hypercholesterolemia, and that relatively low doses decrease LDL cholesterol levels to a greater extent than do similar doses of pravastatin, simvastatin, or atorvastatin as evaluated in separate clinical trials. Pitavastatin, in phase II trials, also has promise as a more potent drug than currently available statins. Because neither of these drugs has been approved for use in the United States, clinical trial results should be considered preliminary. In the future, agents that combine the actions of statins and nicotinic acid may achieve still greater LDL cholesterol reductions. Drugs that lower lipids via mechanisms other than inhibition of HMG-CoA reductase also offer promise. The newest addition to the roster of lipid-regulating agents is ezetimibe, a cholesterol absorption inhibitor that has been approved for use either alone or in combination with a statin. Agents in development include bile acid transport inhibitors and inhibitors of acyl CoA:cholesterol acyltransferase. More research will be needed to determine the full clinical potential of such approaches to the management of hypercholesterolemia. PMID- 12539820 TI - Method of application of tylosin, an antibiotic for American foulbrood control, with effects on small hive beetle (Coleoptera: Nitidulidae) populations. AB - The method of application of the antibiotic tylosin (Tylan) for control of oxytetracycline-resistant American foulbrood (Paenibacillus larvae White) was tested in honeybee (Apis mellifera L.) colonies. A powdered sugar mixture with tylosin, applied as a dust, was efficacious in eliminating American foulbrood symptoms at a rate of 200-mg Tylan per 20 g of powdered sugar, applied at weekly intervals for 3 weeks. A second method of treatment consisting of Tylan mixed with granulated sugar and vegetable shortening and applied once as a patty, at an equivalent total dose as the dust method, to diseased colonies also effectively eliminated symptoms of disease. In all colonies treated with patties, however, small hive beetle (Aethina tumida Murray) populations significantly increased, compared with the powder sugar method or untreated controls. Bee populations in patty-treated colonies also were significantly reduced, most likely the result of the invasion and proliferation of adult and larval small hive beetles. Such reduction in colony strength was not seen in dust-treated colonies. Because of the obvious damaging populations of small hive beetles, concerns about development of disease resistance, unknown risks of residues, and lack of support by regulatory agencies for the use of the patty method, the use of the dust method of tylosin is greatly favored over the patty method. PMID- 12539821 TI - Impact of Macrolophus caliginosus presence on damage production by Dicyphus tamaninii (Heteroptera: Miridae) on tomato fruits. AB - In northeast Spain, the most common predators found in tomato fields and greenhouses are the mirids Macrolophus caliginosus Wagner and Dicyphus tamaninii Wagner. Natural colonization occurs during the growing season and both species prey on whiteflies. Because D. tamaninii can damage tomato fruits during periods of prey scarcity, a semifield experiment was carried out to evaluate whether the presence of M. caliginosus affects damage produced by D. tamaninii. In a tomato greenhouse, 60 plants were individually caged and distinct predator treatments were introduced: D. tamaninii, D. tamaninii + eggs of Ephestia kuehniella Zeller, D. tamaninii + M. caliginosus, M. caliginosus alone, and a control without any insect. Damage to tomato fruits was recorded (>25% of the fruit) in all the treatments with D. tamaninii, whereas no significant damage was detected with M. caliginosus alone. Finally, no intraguild predation was detected between both mirid species. PMID- 12539822 TI - Parasitism of the wheat stem sawfly (Hymenoptera: Cephidae) by Bracon cephi and B. lissogaster (Hymenoptera: Braconidae) in wheat fields bordering tilled and untilled fallow in Montana. AB - We evaluated wheat stem sawfly, Cephus cinctus Norton, parasitism, infestation, and sawfly-cut stems in wheat fields bordering intensely tilled (no visible stubble residue), minimally tilled (>75% stubble residue visible), and untilled (chemical fallow, herbicide fallow management) summer fallow fields in north central and south-central Montana. No difference in sawfly parasitism or sawfly cut stems was found between fields bordering minimally tilled and fields bordering untilled summer fallow. Sawfly parasitism in fields bordering untilled summer fallow was greater than in fields bordering intensely tilled summer fallow at six of the eight sites examined. Sawfly-cut stems were greater in the field bordering intensely tilled fallow at four sites, with no difference in sawfly-cut stems between the intensely tilled and untilled field at the other four sites. Although it has never been reported, we have observed that many sawfly stubs are completely buried. Therefore, we measured the depth of sawfly stubs in four untilled fields in Broadwater County, MT. Two-thirds of the stubs were completely buried (206 of 300) with an average depth of 6 mm. Intensive tillage, which results in soil-covered stubble, is not an effective sawfly control practice, because sawflies typically overwinter below ground and upon emergence must dig to reach the soil surface. However, Bracon cephi (Gahan) and Bracon lissogaster Muesebeck overwinter above ground in stems and might be unable to dig to the soil surface if buried. The elimination of intensive tillage in favor of chemical fallow should result in greater sawfly parasitism over time. Producers replacing minimal tillage with chemical fallow should see no effect on sawfly parasitism. PMID- 12539823 TI - Plant-related factors influence the effectiveness of Neoseiulus fallacis (Acari: Phytoseiidae), a biological control agent of spider mites on landscape ornamental plants. AB - The predatory mite Neoseiulus fallacis (Garman) was evaluated as a biological control agent of herbivorous mites on outdoor-grown ornamental landscape plants. To elucidate factors that may affect predator efficiency, replicated tests were conducted on 30 ornamental plant cultivars that varied in relationship to their generalized morphology (e.g., conifers, shade trees, evergreen shrubs, deciduous shrubs, and herbaceous perennials), production method (potted or field grown), canopy density, and the prey species present on each. Plant morphological grouping and foliar density appeared to be the most influential factors in predicting successful biological control. Among plant morphological groups, N. fallacis was most effective on shrubs and herbaceous perennials and less effective on conifers and shade trees. N. fallacis was equally effective at controlling spider mites on containerized (potted) and field grown plants, and there was no difference in control of mites on plants with Tetranychus spp. versus those with Oligonychus or Schizotetranychus spp. Moderate to unsuccessful control of spider mites by N. fallacis occurred mostly on tall, vertical plants with sparse canopies. Acceptable spider mite control occurred in four large-scale releases of N. fallacis into production plantings of Abies procera, Thuja occidentalis 'Emerald', Malus rootstock, and Viburnum plicatum 'Newport'. These data suggest that N. fallacis can be an effective biological control agent of multiple spider mite species in a range of low-growing and selected higher growing ornamental plants. PMID- 12539824 TI - Predatory role of Neoseiulus fallacis (Acari: Phytoseiidae): spatial and temporal dynamics in Washington red raspberry fields. AB - The seasonal abundance of spider mites and their predator Neoseiulus fallacis (Garman) (Acari: Phytoseiidae) was determined during three consecutive years in Washington State red raspberry fields. Tetranychus urticae Koch (Acari: Tetranychidae), Eotetranychus carpini borealis (Ewing) (Acari: Tetranychidae), and N. fallacis were commonly found in Skagit and Whatcom Counties. E. carpini borealis colonized the fruiting canes earlier in the season than T. urticae. The two phytophages overlapped in midseason, but T. urticae entered diapause earlier than E. carpini borealis and N. fallacis. Densities of N. fallacis increased with increase in spider mite densities. However, the numerical response of the predator was more evident for T. urticae than for E. carpini borealis. Nevertheless, the predator was spatially associated with the two prey species. The spatial and seasonal distribution of N. fallacis in relationship to host plant phenology and prey distribution may influence the effectiveness of this predator as a biological control agent against spider mites in red raspberry. Densities of the predator increased too late to prevent spider mite damage. The predatory role of N. fallacis could be enhanced by introducing or conserving predators that are more tolerant to climatic factors that prevail in and around the cane canopy in the beginning of the season. PMID- 12539825 TI - Developmental rates and host specificity for Pseudacteon parasitoids (Diptera: Phoridae) of fire ants (Hymenoptera: Formicidae) in Argentina. AB - This study extends our comparative knowledge of Pseudacteon interactions with Solenopsis fire ant workers. Reported in this work are development times for seven Argentinean parasitoid species reared on two hosts, Solenopsis richteri Forel and Solenopsis invicta Buren, under laboratory temperature regimes comparable with those of the climatic zones occupied by these host species. Developmental times spanned 31-66 d across phorid species, and in general did not differ between genders or host species, but were longer at lower temperatures. The size distribution of flies reared was bimodal, with a group of large (Pseudacteon borgmeieri, Pseudacteon nocens, Pseudacteon obtusus and Pseudacteon tricuspis) and small (Pseudacteon cultellatus, Pseudacteon curvatus, and Pseudacteon nudicornis) species. P. borgmeieri was exceptional with respect to length of developmental time. Also reported are results of initial oviposition and developmental studies of some of these phorid species on other Argentinean Solenopsis ant species; P. curvatus was the only species able to complete its development on nonhost fire ants. These results support the concept of incorporating several complementary species of Pseudacteon in the biological control of pest fire ants. PMID- 12539826 TI - Life history and cost analysis for continuous rearing of Podisus maculiventris (Say) (Heteroptera: Pentatomidae) on a zoophytophagous artificial diet. AB - The impact of a zoophytophagous, insect-free artificial diet upon the developmental rate, life table parameters, and fertility table parameters was examined over 11 consecutive generations for domesticated and wild colonies of Podisus maculiventris (Say) (Heteroptera: Pentatomidae). This study showed that the developmental time, preoviposition period, fecundity, and nymphal survival improved in the domestic colony when fed an insect-free artificial diet for 11 consecutive generations, but remained relatively constant for the wild colony, as did reproductive rate and intrinsic rate of increase. Although, after 11 generations of adaptation to an artificial diet feeding regime, all reproductive and fertility table parameters were still significantly lower than when fed on T. ni larvae as the natural prey, the realized cost of rearing either colony on the artificial diet approached 1.2 times the cost of rearing these insects on a natural prey. This is a significant achievement in the effort to develop cost effective artificial diets for the mass-rearing of beneficial pentatomids, and has positive implications for the use of one artificial diet to efficiently rear several beneficial insects. PMID- 12539827 TI - Susceptibility of Maconellicoccus hirsutus (Homoptera: Pseudococcidae) to methyl bromide. AB - Eggs, crawlers, early nymphs, late nymphs, and adults of the pink hibiscus mealybug, Maconellicoccus hirsutus (Green), were tested for their susceptibility to methyl bromide in 2-h laboratory fumigations at ambient conditions (25 degrees C, 95% RH). Dose-response tests indicated that the egg was the most susceptible life stage with an LC99 of 20.2 mg/liter. Based on probit analysis of dose response data, no significant differences were observed among susceptibilities of the crawler, early stage or late stage nymphs, or adults at either the LC50 or LC99 level, but late stage nymphs were more tolerant than early stage nymphs in a separate paired comparison test. Confirmatory tests showed that a dose of 48 mg/liter methyl bromide, the USDA-Animal Plant Health Inspection Service treatment dose schedule for mealybugs at 21-26 degrees C, produced 100% mortality of all life stages. On the basis of these results, we conclude that the methyl bromide treatment schedule for mealybugs will provide quarantine security for M. hirsutus infesting commodities for export or import. PMID- 12539828 TI - Citrus pollen retention by adult Helicoverpa zea (Lepidoptera: Noctuidae) after exposure to citrus blooms. AB - In previous migration studies, the presence of citrus pollen on adult Helicoverpa zea (Boddie), in conjunction with synoptic weather systems and 72-h backtrack trajectories, were used to identify source zones of migrants. However, data are lacking regarding the retention of citrus pollen for 72-h by H. zea adults. We exposed laboratory-reared and feral H. zea adults to citrus blooms for a 12-h period in laboratory and field studies and examined insects for the external presence of citrus pollen (i.e., marking) at 12-h intervals through 72 h. Citrus pollen marking was higher for females than males at the time of removal from citrus blooms. Fifteen to 100% of H. zea adults were marked with citrus pollen at 72 h after removal from citrus blooms. Pollen loads ranged from rare (< or = 10 pollen grains) to moderate (101 to 500 grains) during 1995; only rare and light (11 to 100 grains) pollen loads were detected during 1996. Citrus pollen marking of H. zea adults through 72 h after removal from citrus blooms has not been previously confirmed. These data provide evidence of H. zea using available blooming citrus groves in the absence of a host crop and will impact current perspectives regarding H. zea and host plant interactions and timing of pest management tactics. PMID- 12539829 TI - Seasonal population dynamics of the greenhouse whitefly Trialeurodes vaporariorum (Homoptera: Aleyrodidae) on strawberries in Southern California. AB - The greenhouse whitefly, Trialeurodes vaporariorum (Westwood), has recently become a major insect pest of strawberries in Southern California. The population dynamics of this pest were monitored over 2 yr in six commercial strawberry fields near the coastal communities of Oxnard and Ventura under two crop production regimes, summer- and fall-planted strawberries. Adult whitefly numbers generally peaked during the February through May period for fall-planted strawberries and during the October through November period for summer-planted strawberries. Population densities varied greatly among fields within each regime and the differences were likely caused by surrounding alternate host crops. PMID- 12539830 TI - Toxicities of emamectin benzoate homologues and photodegradates to Lepidoptera. AB - The toxicity of a number of emamectin benzoate homologues and photodegradates to five species of Lepidoptera was investigated using diet and foliar bioassays. The emamectin benzoate homologues B1a and B1b were equally toxic in the diet and foliar assays to Spodoptera exigua (Hubner), Heliothis virescens (F.), Tricoplusia ni (Hubner), and Spodoptera frugiperda (J. E. Smith), within each of these species. Plutella xylostella (L.) was the most sensitive species to emamectin benzoate. The AB1a photodegradate of emamectin benzoate was as toxic as the parent compound in the diet assay. However, in the foliage assay AB1a was 4.4 fold less toxic to S. exigua than the parent compound. The MFB1a photodegradate of emamectin benzoate was as toxic as the parent compound to P. xylostella, and 3.1 to 6.2 times as toxic as the parent compound to the other species in the diet assay. The order of toxicity of the photodegradates were AB1a > MFB1a > FAB1a > 8,9-Z-MAB1a > PAB1a. PMID- 12539831 TI - An inexpensive, accurate method for measuring leaf area and defoliation through digital image analysis. AB - We report a protocol using a common desk-top scanner and public domain software for measuring existing leaf area and leaf area removed as a result of herbivory. We compared the accuracy and precision of this method to that of a standard leaf area meter. Both methods were used to measure metal disks of a known area, the area of soybean (Glycine max L.) leaves, and the area removed by simulating leaf feeding with a hole-punch. We varied the amount of injury across a low, medium, and high degree of simulated feeding. The mean area of 10 cm2 and 50 cm2 metal disks was more accurately estimated with the leaf area meter than the desk-top scanner. Leaf area estimates from both methods were highly correlated. The desk top scanner accurately estimated the leaf area removed from the low, medium, or high degree of simulated leaf feeding. However, the leaf area meter overestimated low levels of simulated feeding injury. Though measuring a leaf's surface area with a desk-top scanner requires two steps (creating a digital image file and calculating the area represented by that image), the overall time required to measure leaf injury is shorter than with a leaf area meter. This relatively simple and inexpensive method of estimating leaf area and feeding damage has advantages in certain experimental situations where a prefeeding measurement of the leaf is impossible or undesirable, or when small amounts of feeding occur. PMID- 12539832 TI - Tomicus piniperda (Coleoptera: Scolytidae) initial flight and shoot departure along a north-south gradient. AB - The exotic pine shoot beetle, Tomicus piniperda (L.) (Coleoptera: Scolytidae), established in the north central and northeastern United States (U.S.) and adjacent regions in Canada, is regulated by a federal quarantine that restricts movement of pine material during specific times of the year based on the beetle's life history. Although climatic variation occurs across T. piniperda's range, a single set of dates is used for timing the movement of pine logs. We monitored T. piniperda spring flight, fall shoot departure, and air and internal tree temperatures at three sites along a 300-km north-south gradient in Michigan and Indiana. We also estimated dates for initial spring flight (12 degrees C threshold) and fall shoot departure (0 degrees C threshold) across an 850-km gradient using historical temperature records (1901 to 1999). Average daily temperatures in fall 1997 (8 October to 12 December) and spring 1998 (20 February to 21 April) were 1.8 to 2.4 degrees C colder, respectively, at the northern field site than at the southern field site. Fall shoot departure began at approximately the same time (day 289 to 290) at all three field sites, but complete shoot departure was extended by 3 wk at the southern site (day 336) compared with the northern site (day 317). T. piniperda adults were first captured in funnel traps on calendar day 86 at the northern site and on day 59 at the central and southern field sites. Peak flight occurred at approximately the same time (day 86) at all three sites. Within-shoot temperatures were very similar to air temperatures in the fall and aboveground inside-bark temperatures were similar to air temperatures in the spring. Average predicted dates based on historical temperature records varied by 31 d for initial shoot departure and 84 d for initial spring flight between northern Michigan and southern Indiana. Because considerable variation can occur in T. piniperda behavior across a broad geographic range, dates specified in the U.S. Federal quarantine should be adjusted according to local temperatures. PMID- 12539833 TI - Evaluation of preventive treatments in low-density gypsy moth populations using pheromone traps. AB - Pheromone traps can be used for evaluating the success of treatments that are applied to either eradicate or delay the growth of isolated low-density populations of the gypsy moth, Lymantria dispar (L.). We developed an index of treatment success, T, that measures the reduction in moth counts in the block treated adjusted by the change in moth counts in the reference area around it. This index was used to analyze the effectiveness of treatments that were conducted as part of the USDA Forest Service Slow-the-Spread of the gypsy moth project from 1993 to 2001. Out of 556 treatments that were applied during this period, 266 (188,064 ha) were selected for the analysis based on several criteria. They included 173 blocks treated with Bacillus thuringiensis (Berliner) variety kurstaki and 93 blocks treated with racemic disparlure. Analysis using general linear models indicated that disparlure treatments were significantly more effective than B. thuringiensis treatments in reducing moth captures. The frequency of repeated treatments in the same area was higher after B. thuringiensis than after disparlure applications. Treatments were more successful if the pretreatment moth counts outside of the block treated were low compared with moth counts inside the block. PMID- 12539834 TI - Population dynamics of Frankliniella spp. and tomato spotted wilt incidence as influenced by cultural management tactics in tomato. AB - We investigated the effects of ultraviolet (UV)-reflective mulch and two rates of nitrogen fertilization on populations of Frankliniella spp. thrips and on the incidence of tomato spotted wilt in field-grown tomato in northern Florida. The higher of the two soil nitrogen fertilizer treatments significantly increased populations of Frankliniella occidentalis (Pergande), whereas mulch reflectance significantly decreased populations of F. occidentalis. Populations of Frankliniella tritici (Fitch) were decreased only by UV-reflective mulch. Decreased thrips populations in UV-reflective mulch plots were probably due to disruptions in host-finding behavior. Increased thrips populations in tomatoes treated with the higher nitrogen fertilization rate were probably due in part to increased nutrients available in flowers. Incidence of tomato spotted wilt was significantly decreased in tomatoes grown on UV-reflective mulch, whereas disease incidence was significantly greater in increased nitrogen-fertilized plots. This research reveals that cultural practices resulted in up to 45% reduction in the numbers of vector and nonvector species of flower thrips and up to 50% reduction in tomato spotted wilt. PMID- 12539835 TI - Preference of food particle size among several urban ant species. AB - Appropriate particle size may be a critical characteristic for effective granular ant baits. We examined the particle size preference of six species of pest ants to an anchovy-based bait. We also examined head capsule widths of Argentine ants, Linepithema humile (Mayr) (mean = 0.54 mm), California harvester ants, Pogonomyrmex californicus (Buckley) (mean = 1.63 mm), red imported fire ants, Solenopsis invicta Buren (mean = 0.9 mm), and southern fire ants, Solenopsis xyloni McCook (mean = 0.76 mm) and compared them with the first and second most preferred particle size. There were differences between particle size of which the most mass was removed and of which there were more particles removed by ants. California Argentine ants, southern fire ants, and Alabama Argentine ants removed more 840 to 1,000-microm particle mass of the anchovy diet but had more visits to dishes containing 420 to 590 microm particles. California harvester ants and Allegheny mound ants, Formica spp., removed more >2,000 microm particle mass but visited dishes containing 1,000 to 2,000 microm particles more often. Red imported fire ants also removed more >2,000 microm particle mass but visited dishes with 590 to 840-microm particles most often. Pharaoh ants, Monomorium pharaonis (L.), removed and visited 420 to 590-microm particles more than any other size. A linear regression model determined that particle size preferred by each ant species relates to forager head width. The majority of particles of commercial ant bait, including Amdro, Ascend, Award, Bushwhacker, Max Force with fipronil, and old and new formulations of Max Force with hydramethylnon, were 1,000 to 2,000 microm, but the majority of Niban particles were <420 microm. Altering the size of particles of toxic ant baits to fit the particle size preference of each pest ant species may increase the efficacy of ant baits. PMID- 12539836 TI - Negative correlations between resistance to three organophosphate insecticides and productivity within a natural population of Drosophila melanogaster (Diptera: Drosophilidae). AB - To investigate the relationship between resistance to organophosphate insecticides and fitness components, we first measured resistance to three organophosphates, malathion, prothiophos, and fenitrothion, and productivity, a measure of fitness components, for each of the isofemale lines from the same natural population of Drosophila melanogaster (Meigen). Pearson correlation coefficients indicated that positive correlations among resistance to the organophosphates and negative correlations between resistance to each of the organophosphates and the productivity existed within the natural population. We further investigated the genetic basis of the correlations among resistance to the organophosphates and the productivity, by using chromosome-substituted lines between a resistant and a susceptible inbred line established from the same natural population. Chromosomal analyses indicated that the third chromosome from the resistant line exhibited not only significant, positive effects on resistance to all of the organophosphates tested but also a significant negative effect on the productivity, suggesting positive genetic correlations between resistance to each organophosphate and negative genetic correlations between resistance to each organophosphate and the productivity. In addition, a significant negative effect on the productivity was also detected from the second chromosome, which did not exhibit significant major effects on resistance to the organophosphates. This suggests that fitness components of resistant lines could be also affected by factors independent of insecticide resistance. The dynamics of genetic variation in resistance to the organophosphates within the natural population of D. melanogaster are discussed from the standpoint of negative genetic correlations between resistance to the organophosphates and the productivity. PMID- 12539837 TI - Comparative inhibition kinetics for acetylcholinesterases extracted from organophosphate resistant and susceptible strains of Boophilus microplus (Acari: Ixodidae). AB - In this study, acetylcholinesterases (AChEs) were extracted from two Mexican Boophilus microplus strains that demonstrated resistance to the organophosphate (OP) acaricide, coumaphos, in bioassay. The rate of inhibition of the extracted AChEs by the diethyl-OP paraoxon was determined for two resistant strains and two susceptible strains of B. microplus. The time to inhibition of 50% AChE activity was approximately two-fold greater for the resistant strains. Kinetic analysis of the interaction of the resistant AChEs with paraoxon revealed reduced bimolecular reaction constants (ki). Apparent conformational changes in the AChE of the resistant strains were reflected in reduced Km and Vmax values. The bimolecular reaction constants (ki) of the resistant strains were most affected by a slower rate of enzyme phosphorylation (k2). PMID- 12539838 TI - Impact of an insecticide resistance strategy for house fly (Diptera: Muscidae) control in intensive animal units in the United Kingdom. AB - A strategy for house fly (Musca domestica L.) control in intensive animal units in the United Kingdom was proposed by the Pesticide Safety Directorate (PSD) of the Ministry of Agriculture, Fisheries and Food in 1993. An advice leaflet was circulated to farmers, and label recommendations for insecticides used to control house flies were altered to prevent their long-term and frequent use. A study was carried out between 1996 and 1998 to gather data on insecticide use and resistance in house fly populations and compared with results from a study carried out in 1990-1992 to assess the impact of the 1993 label recommendations. As in the 1990-1992 study, resistance to methomyl, azamethiphos and pyrethrins + piperonyl butoxide was assessed. Larvicide tests with cyromazine, which had recently been released in the United Kingdom, were also included in this study. Most of the farmers claimed to have received and read the PSD insecticide advice leaflet, and half claimed to have altered insecticide treatments as a result. Comparing results for insecticides used before and after 1993, the proportion of farmers claiming to have used each of the insecticides had decreased. However, there had been no amelioration in resistance to synergised pyrethrins, and the number of house fly populations with reduced response to the insecticide baits had increased between 1990-1992 and 1996-1998. All the house fly populations tested were fully susceptible to cyromazine. There is an urgent need, therefore, to devise new strategies and particularly to minimize the risk of selecting for resistance to cyromazine. PMID- 12539839 TI - Genetics and fitness costs of cyromazine resistance in the house fly (Diptera: Muscidae). AB - The genetic basis of cyromazine resistance was investigated in the house fly, Musca domestica L. The ED-R strain, which was collected in Mississippi and selected further in the laboratory, was 116.5-fold resistant compared with the laboratory susceptible strain, OR-S. The SEL strain, which was created by crossing ED-R with OR-S followed by three cycles of reselection and backcrossing to OR-S, was 84.7-fold resistant relative to the susceptible strain. Mortality data from reciprocal crosses of resistant and susceptible flies indicated that resistance was autosomal and not influenced by maternal effects. The relative position of probit lines from the parental strains and reciprocal crosses showed that resistance was expressed as an incompletely dominant trait with D = 0.30 and 0.32 for ED-R and SEL, respectively. To determine the number of genes involved, models of one, two, three, four, and five loci were used to compare observed and expected mortality of F1ED-R x susceptible backcross. Resistance was best described by a polygenic model of three loci when equal and additive effects of loci were assumed. Another approach, which was based on phenotypic variances, showed that nE, or the minimum number of freely segregating genetic factors for ED-R, equaled 3.07. ED-R showed greater reductions in fitness compared with SEL independent of the presence or absence of sublethal concentrations of cyromazine. These data suggested that reduced fitness was not due to deleterious pleiotropic effects of the resistance genes themselves but arose from other loci in the ED-R genotype. PMID- 12539840 TI - Diagnostic assays based on esterase-mediated resistance mechanisms in western corn rootworms (Coleoptera: Chrysomelidae). AB - Resistance to methyl-parathion among Nebraska western corn rootworm, Diabrotica virgifera virgifera LeConte, populations is associated with increased hydrolytic metabolism of an organophosphate insecticide substrate. An electrophoretic method to identify resistant individuals based on the staining intensity of esterase isozymes on nondenaturing polyacrylamide gels was developed. Three groups of esterases (I, II, and III) were visible on the gels, but only group II esterase isozymes were intensified in resistant populations. A total of 26 and 31 field populations of western corn rootworms from Nebraska (in 1998 and 1999, respectively) were assessed with nondenaturing polyacrylamide gel electrophoresis (PAGE) assays and diagnostic concentration bioassays. Significant correlations were observed between the two diagnostic assays. Group II esterase isozymes provide a reliable biochemical marker for detection of methyl-parathion resistance in individual western corn rootworms and a tool for monitoring the frequency of resistant individuals in field populations. PMID- 12539841 TI - Genetic basis of resistances to chlorfenapyr and etoxazole in the two-spotted spider mite (Acari: Tetranychidae). AB - We studied the genetic basis of resistance to two new acaricides, chlorfenapyr and etoxazole, which have different chemical structures and modes of action in the two-spotted spider mite, Tetranychus urticae Koch. The resistance ratios calculated from the LC50s of resistant and susceptible strains were 483 for chlorfenapyr and >100,000 for etoxazole. Mortality caused by the two acaricides in F1 progeny from reciprocal crosses between the resistant and susceptible strains indicated that the modes of inheritance of resistance to chlorfenapyr and etoxazole were completely dominant and completely recessive, respectively. Mortality in F2 progeny indicated that for both acaricides, the resistance was under monogenic control. Repeated backcross experiments indicated a linkage relationship among the two acaricide resistances and malate dehydrogenase, although phosphoglucoisomerase was not linked with them. The recombination ratio between the resistances was 14.8%. From this result, we suggest that heavy spraying of the two acaricides will lead to apparent cross-resistance as a consequence of crossing over; the two resistance genes are so close to each other that it would be difficult to segregate them once they came together on the same chromosome. PMID- 12539842 TI - Suitability of transgenic glyphosate-resistant soybeans to green cloverworm (Lepidoptera: Noctuidae). AB - Host plant suitability to green cloverworm, Hypena scabra (F.), was evaluated on two conventional soybean varieties (Stine 2499-0 and 2972-2) and two RoundUp Ready Soybeans (Stine 2506-4 and 2892-4) with and without exposure to glyphosate. No differences among treatments were detected on developmental time and survivorship. Developmental time from first instar to adult ranged from 24.7 to 25.5 d, and survivorship ranged from 86 to 96%. No sex bias was observed among treatments (proportion of females ranged from 0.41 to 0.50). Morphological differences were observed between sexes; males had a longer and wider thorax, longer wings, and longer body. Females had longer and wider abdomens. Although treatments did not affect size (calculated with principal component analyses), significant differences were observed between males and females. These results suggest that soybean genetic differences (between conventional varieties and analogous transgenic varieties) or plant stress (induced by glyphosate metabolization) do not affect the plant suitability to H. scabra. PMID- 12539843 TI - Resistance in accessions of cowpea to the coreid pod-bug Clavigralla tomentosicollis (Hemiptera: Coreidae). AB - Laboratory and screenhouse experiments were conducted to identify antibiosis and tolerance in four wild accessions of cowpea, Vigna unguiculata subsp. dekindtiana, to Clavigralla tomentosicollis Stal, the most damaging pod-sucking bug on cultivated Vigna genotypes in West Africa. The wild accession TVnu 151 showed antibiosis resistance, causing >50% mortality of the nymphs within 3 d of placing them on pods. Nymphs died more quickly on TVnu 151 than on TVnu 72, the wild and resistant control of the V. vexillata species which affected the weights of surviving insects to a much greater degree than TVnu 151. The three other accessions of the subspecies dekindtiana (TVnu 369, TVnu 517, and TVnu 707) did not cause significant mortality to the bugs, but rather extended their developmental time, with surviving adults showing lower weights and slower oviposition rates than those on the susceptible control IT84S-2246. Wild accessions affected male and female bugs differently, resulting in differential survival. This was reflected by the sex ratio which was male biased on the wild accessions (1:0.3-1:0.9), and female biased on IT84S-2246 (1:1.5). No evidence of tolerance was found in the four accessions of V. unguiculata subsp. dekindtiana. Overall, seed traits seemed to be the major resistance component in these wild accessions. PMID- 12539844 TI - Chemical stimuli from corn plants affect host selection and oviposition behavior of Sesamia nonagrioides (Lepidoptera: Noctuidae). AB - The role of chemical stimuli in host selection and oviposition of Sesamia nonagrioides (Lef) was studied in the laboratory using eight corn hybrids. Female oviposition preference was evaluated on plants of the eight hybrids, their extracts in pentane and methanol, and volatiles collected by steam distillation in ether. Results from two-choice bioassays indicate various degrees of female preference to the hybrids. Four hybrids received more eggs than control plants, whereas the remainder received fewer eggs. Relatively similar results were obtained when filter paper sticks baited with pentane and steam distillation extracts were evaluated. Methanol extracts from all eight hybrids received fewer eggs than the methanol controls. Results showed that pentane and steam distillation extracts contain chemicals that elicit female oviposition response, while methanol extracts may contain chemicals that deter or repel females to oviposition. PMID- 12539845 TI - Survival and development of Heliothis virescens (Lepidoptera: Noctuidae) larvae on isogenic tobacco lines with different levels of alkaloids. AB - Levels of pyridine alkaloids were measured in 18 tobacco, Nicotiana tabacum L., entries from three parental isolines ('NC 95', 'SC 58', and 'Coker 139'), grown at Tifton, GA, Florence, SC, and Oxford, NC, in 1991. Levels of alkaloids in bud leaves (first fully unfolded leaf below the apical leaf bud) were negatively correlated to natural infestation ratings of tobacco budworm larvae, Heliothis virescens (F.), 7 wk after transplanting. For artificially infested bud leaves at Oxford, there was a significant negative correlation between levels of total alkaloids and larval weights after 1 wk of feeding. In 1992, four entries from the 'NC 95' isoline were grown at Oxford, and samples for alkaloid analyses were taken every 2 wk at several leaf positions on each plant. During weeks 4, 8, 12, and 16, second instar tobacco budworms were caged on individual, intact leaves inside perforated plastic bags in the field. The survival and development of tobacco budworm larvae after 1 wk were negatively correlated with levels of alkaloids at the various leaf positions. Larvae survived better and grew faster on the bud leaves of each entry where alkaloid levels were lower than they did on leaves further down the stalk where alkaloid levels were higher. More larvae survived on the lower leaves of the low alkaloid lines than on the lower leaves of the high alkaloid lines. Even moderate increases in pyridine alkaloids had negative effects on tobacco budworm survival and development. Nicotine constituted >97% of the pyridine alkaloids in the 'NC95' isoline each year. PMID- 12539846 TI - Effect of husk characters on resistance to corn earworm (Lepidoptera: Noctuidae) in high-maysin maize populations. AB - Two maize (Zea mays L.) breeding populations with very high concentrations of maysin, a silk-expressed flavone glycoside, were tested for their ability to resist ear damage by the corn earworm, Helicoverpa zea Boddie, under field conditions. Tests were conducted in 2000 and 2001 at multiple locations in Georgia. The high maysin populations, EPM6 and SIM6, as well as resistant and susceptible checks, were scored for silk-maysin content, H. zea damage, and husk characters. In 2000, there was a negative correlation between husk tightness and earworm damage at three of five locations, while there was no significant correlation between damage and maysin content at any location. In 2001, EPM6 and SIM6 had approximately ten times the maysin content of the low-maysin control genotypes; nevertheless, earworm damage to EPM6 and SIM6 was either greater than or not significantly different from the low-maysin genotypes at all locations. The resistant control genotype, Zapalote Chico, had significantly less earworm damage than EPM6 and SIM6 for both years at all locations. The results of this study highlight the importance of identifying and quantifying husk and ear traits that are essential to H. zea resistance in maize. PMID- 12539847 TI - Head capsule width as an instar indicator for larvae of the cranberry fruitworm (Lepidoptera: Pyralidae) in southeastern New Brunswick. AB - The objective of this study was to determine the number of instars of the cranberry fruitworm Acrobasis vaccinii Riley in southeastern New Brunswick based on the distribution of head capsule widths from field and laboratory observations. In 2000, head capsules from field samples were measured across their widest point, and the results were plotted against observed frequencies. The data from field samples suggested that A. vaccinii exhibited five instars in 2000. In 2001, larvae were reared in the laboratory until the final molt, and head capsules were counted and measured. The results were also plotted against observed frequencies. None of the laboratory specimens exhibited more than five instars, supporting the results of the previous year. Various factors are invoked to explain the difference between these results and those of a previous study conducted 50 yr earlier and 200 km away, in which six instars and larger head capsules were reported. PMID- 12539848 TI - Efficacy of ethiprole applied alone and in combination with conventional insecticides for protection of stored wheat and stored corn. AB - The insecticidal pyrazole ethiprole, applied at rates of 7.5 and 10.0 ppm either alone or in combination treatments with deltamethrin, piperonyl butoxide, and chlorpyrifos-methyl, was evaluated as a protectant of stored wheat and stored corn. The commodities were treated with six treatment combinations, including an untreated control, and held for 6 mo at 22, 27, or 32 degrees C and 57% RH. Bioassays were conducted monthly by exposing the rice weevil, Sitophilus oryzae (L.), and the red flour beetle, Tribolium castaneum (Herbst), on treated wheat and the maize weevil, Sitophilus zeamais (Motschulsky), and the red flour beetle on treated corn. The storage temperature of wheat did not significantly affect mortality of exposed insects (P > or = 0.05). All rice weevils were dead after 1 wk in all treatments, and no F1 adults were produced. Mortality of red flour beetles was not dependent on either chemical treatment or bioassay month, and no F1 adults were produced. The storage temperature of corn did not significantly affect mortality of exposed insects (P > or = 0.05). Mortality of maize weevils varied from 77.9 to 100% in all chemical treatments, and no F1 adults were produced. Mortality of red flour beetles was also variable among treatments and bioassay month and no F1 adults were produced. This is the first published report of a study in which pyrazoles have been evaluated against stored-grain insects. PMID- 12539849 TI - Age estimation of Mexican fruit fly (Diptera: Tephritidae) based on accumulation of pterins. AB - A common method of aging adult flies, fluorescence spectrometry, was used to monitor the increase of overall pterine titer in head extracts of Anastrepha ludens (Loew). Accumulation of fluorescent compounds was measured as a function of chronological age of flies maintained at 17 and 27 degrees C. Although relative fluorescence increased with age, field studies revealed that this phenomenon could not be used for accurate age estimation, as relative fluorescence did not increase predictably with age over the entire life span. Accumulation of individual pterins, deoxysepiapterin and sepiapterin, were studied in a similar manner. These two specific compounds were separated by high pressure liquid chromatography and their accumulation was followed at 15 and 30 degrees C in the laboratory and under caged field conditions. While titer of deoxysepiapterin increased steadily in a curvilinear fashion, sepiapterin quickly reached a maximum and then maintained a constant level for the rest of the life of the flies. Based on the physiological response of deoxysepiapterin to chronological time and ambient thermal conditions, this compound was determined to be an age specific biological parameter for the Mexican fruit fly and should allow age estimation in field-collected flies. PMID- 12539850 TI - Hole density and capture of stored-product insect pests in grain probe traps. AB - The relationship between number of holes in a grain probe trap body and capture of stored-grain pests was determined in laboratory tests using adults of the rice weevil, Sitophilus oryzae (L.), the sawtoothed grain beetle, Oryzaephilus surinamensis (L.), and the red flour beetle, Tribolium castaneum (Herbst). Polyvinylchloride (PVC) probe bodies were attached to electronic sensor heads, and insect captures were recorded electronically using an Electronic Grain Probe Insect Counter (EGPIC) system. In comparisons among PVC probe trap bodies with 60 492 holes, tested at 71 insects per kg in 2.8 kg of soft wheat in cylindrical mini-silos, sawtoothed grain beetle and rice weevil captures were directly related to number of holes in the probe trap body, but there was no relationship for red flour beetle capture. Subsequent tests were conducted comparing sawtoothed grain beetle and rice weevil captures in a PVC probe body with 210 holes over a 40-cm long trapping surface with two commercially available probe traps, a polycarbonate (Lexan) probe trap with 180 holes over a 14-cm long trapping surface and a polyethylene (WBII) probe trap with 750 holes over a 34-cm long trapping surface. The highest percentage capture of both species was in the WBII probe trap, but the 210-hole PVC probe body was as effective as the Lexan probe body for rice weevils and sawtoothed grain beetles at 71 and 17 insects per kg of wheat, respectively. PMID- 12539851 TI - Mechanisms underlying sawtoothed grain beetle (Oryzaephilus surinamensis [L.]) (Coleoptera: Silvanidae) infestation of consumer food packaging materials. AB - The sawtoothed grain beetle, Oryzaephilus surinamensis (L.), is an extremely destructive pest of packaged consumer food products. The beetle is not believed to chew directly through packaging materials, but to use openings or flaws in damaged or improperly sealed packages to gain entry. We investigated the behavioral mechanisms by which the sawtoothed grain beetle infests packages with flaws. Significantly more sawtoothed grain beetles infested consumer food packages that had been punctured with 0.4 mm diameter holes, to simulate packaging flaws that preclude adults, than when packages had no flaws. In a test arena, females laid more eggs into or near the hole in a plastic packaging film, when they were able to contact the food through the hole than when they could not contact the food. First instar larvae placed either 1 mm or 1 cm away entered holes when food was present, indicating that packages could become infested if eggs were laid near holes. In the absence of food, neither adults nor larvae responded to holes. This study has shown the importance of sound packaging in preventing insect infestation. PMID- 12539852 TI - Recycled paper products as substitutes for gelling agent in screwworm (Diptera: Calliphoridae) larval diet. AB - Absorbent recycled paper products were tested and compared with a sodium polyacrylate gelling agent, Aquatain, for solidifying larval diets of the screwworm, Cochliomyia hominivorax (Coquerel). A recycled newsprint product, Terra-Mulch, when mixed with water and dietary ingredients produced a diet medium of homogeneous texture that supported larval growth and development comparable to the standard diet prepared with the gelling agent. Biological yields as calculated from number of pupae per tray from both the diets were similar. Three other types of recycled paper products were tested. Although less homogeneous in texture, these diets were as good as the diet with Terra-Mulch. There were no significant differences in the mean larval and pupal weights, no. of pupae/tray, percent emergence, oviposition, percent hatch, and percent longevity. When tests were conducted through eight consecutive generations, no adverse effect was noticed in any of the life history parameters. Terra-Mulch is inexpensive and biodegradable compared with the gelling agent Aquatain that is expensive and not biodegradable. Replacing Aquatain with Terra-Mulch in the screwworm larval diet for mass rearing may result in an annual savings of more than half a million U.S. dollars in costs of material and labor. PMID- 12539853 TI - The influence of ambient room lighting on the pattern electroretinogram (PERG). AB - It has been suggested that low ambient lighting conditions increase the amplitude of the PERG, but no data has been available on this issue. We recorded the transient PERG (0.8 degrees check size) and steady-state PERG (15 rev/s, 0.8 degrees and 16 degrees check size) under three lighting conditions: dark room, only illuminated by the stimulus (resulting in 30 lux), our standard room lighting (windows occluded, one lighted lamp, 200 lux) and fully lit room (full ceiling illumination with eight fluorescent tubes) resulting in rather bright 2300 lux. The stimulus luminance was 50 cd/m2. The sequence of lighting conditions varied for each subject and followed a balanced permutation of an ABCCBA scheme. Results showed a significant effect (P < 0.01) across lighting conditions, with no relevant difference between the 30 and 200 lux conditions, but a reduction down to 70% at the 2300 lux condition. This obtained across all check sizes and temporal conditions. As an example, the transient PERG P50 amplitudes were as follows: dark, 5.6 +/- 0.8 microV; medium, 5.3 +/- 0.6 microV and bright, 3.8 +/- microV (mean +/- SEM). Peak times decreased significantly with illumination (dark, medium or bright): 45.9 +/- 0.9, 43.1 +/- 0.6 or 40.8 +/ 0.8 ms. Contrast measurements quantitatively explained the noticeable reduction of PERG amplitude at the brightest illumination level simply by straylight, which reduced the display contrast. This suggests that bright sunlight should be excluded, and that lighting conditions should be moderately standardized at low or medium luminance levels for reproducible amplitudes and peak times. PMID- 12539854 TI - Cranial hypertension as first manifestation of Behcet's disease: a case report. AB - Behcet's disease (BD) is a chronic, multisystem and relapsing vasculitis of unknown etiology. Central nervous system (CNS) involvement is reported in 30% of cases, but it is the first symptom of the disease in only 5% of subjects. Neurological manifestations may appear as a parenchymal CNS pattern (the commonest), an intracranial hypertension-like pattern, or a meningitis-like pattern. We describe a 30-year-old Algerian man with BD who developed, as first symptom, a typical intracranial hypertension picture with headaches, bilateral papilledema and raised cerebrospinal fluid (CSF) pressure. Magnetic resonance angiography (MRA) revealed a cerebral venous sinus thrombosis (VST). After 1 month of treatment with anticoagulants, prednisone, colchicine and chlorambucil, MRA showed complete recanalization of the cerebral venous sinus and the patient made a full recovery. BD should be routinely looked for in adult patients, especially males in their third and fourth decades, who present with intracranial hypertension syndrome. We therefore advocate the use of MRA for unexplained neurological symptoms in BD, since without it cerebral VST may easily be missed. PMID- 12539855 TI - Long-term therapy with low dose cyclosporin A in ocular Behcet's disease. AB - Ocular complications of Behcet's disease can be severe and lead to blindness in 90% of untreated patients. We aimed to evaluate the long-term use of low dose cyclosporin-A (CsA) which is a potent immunomodulatory agent in the treatment of ocular Behcet's disease. Fifty-two patients (104 eyes) with ocular Behcet's disease using CsA for at least 1 year were included in this study. All the patients underwent complete ophthalmological and systemic examination. Five mg/kg/per day CsA was started to the patients with severe posterior uveitis and/or frequent anterior inflammatory attacks unresponsive to the conventional therapeutic agents. According to ocular response and adverse effects, the dose was tapered gradually over 2 months to a maintenance dose of 3 mg/kg/per day. Prednisone (0.2-0.8 mg/kg/per day) was added when necessary. Forty-six of the patients (88.5%) were males and six (11.5%) were females. The mean age was 33.65 +/- 7.75 (range, 19-53) years. The mean ocular involvement period was 64.1 (range, 12-180) months. Posterior uveitis was present in 49 (94.21%) and severe, recurrent anterior uveitis in three (5.8%) patients. The mean CsA administration period was 38 +/- 18.1 months. Visual acuity improved in 31(29.8%), deteriorated in 32 (30.8%) and unchanged in 41(39.4%) of the 104 eyes. No ocular attacks occurred in 50% of the eyes during therapy. Nine (17.3%) of the patients had to stop the therapy because of the adverse effects of the CsA and the others tolerated well for a long-term period. CsA is not the ideal therapeutic agent in ocular Behcet's disease because it can not completely eliminate the disease, but it is currently one of the most effective and efficient drug to control the uveitis and its complications until better treatment modalities are developed. PMID- 12539856 TI - The effects of GABA and vigabatrin on horizontal cell responses to light and the effect of vigabatrin on the electroretinogram. AB - When used as an antiepileptic drug in humans vigabatrin, which is a GABA analogue and an inhibitor of GABA-aminotransferase, often causes peripheral visual field loss. This could result from increases in endogenous GABA levels. Accordingly we have investigated the effects of GABA on horizontal cells (HCs) of the rabbit retina, and of vigabatrin, when applied for only a few minutes, on HCs and on the electroretinogram (ERG). The intracellular HC and ERG records were first obtained from isolated rabbit retinas during perfusion with a physiological solution. The perfusate was then changed to one containing GABA (2 mM) or vigabatrin (25, 40 or 150 microM) for at least 5 min, and then returned to the control solution. 2 mM GABA significantly but reversibly reduced the light responses of HCs elicited by diffuse light (at -4 log intensity) to 52 +/- 17% (SD, n = 7). Vigabatrin had no significant effect on the light responses of HCs (n = 7), and no effect on the b wave (n = 4), but the PIII-component of the ERG was slightly but significantly reduced to 84 +/- 5% (SD, n = 5). The high dosage of GABA needed to affect the light responses of HCs could be due to strong GABA uptake systems in the intact rabbit retina. It is, however, possible that in humans receiving long-term treatment with vigabatrin, high levels of GABA occur because of the inhibition of GABA- aminotransferase. It seems, from these observations, that neurons like on bipolar cells, which are contributors to the b-wave, and HCs are uninfluenced by vigabatrin in short-term experiments. The slightly reduced slow PIII-component, however, indicates an influence on the glial Muller cells which are the main contributors to the slow PIII-component. PMID- 12539857 TI - Nitric oxide and octreotide in retinal ischemia-reperfusion injury. AB - This experimental study was performed to investigate the role of ischemia reperfusion injury on retinal nitric oxide activity and to determine whether octreotide, the synthetic analogue of natural somatostatin, modifies the nitric oxide activity during retinal ischemia-reperfusion in a quinea pig model. Three groups of seven pigmented male quinea pigs were formed; Control, Ischemia and the Ischemia/Octreotide groups. 90 minutes of pressure-induced retinal ischemia and 24 h of reperfusion were established in the ischemia and ischemia/octreotide groups. Saline for the ischemia group and 50 microg/kg of octreotide for the ischemia/octreotide group were administered intraperitoneally five times with 6-h intervals. At the end of the reperfusion period both eyes of the animals of the three groups were enucleated. One eye of each animal was randomly selected for biochemical assay and the other for histopathological analysis. Retinal nitrate levels were measured and histopathological changes were evaluated in the groups. The mean retinal nitrate levels of the control, ischemia and ischemia/octreotide groups were 157.6 +/- 25.2, 106.4 +/- 20.1 and 96.4 +/- 17.7 micromol/l, respectively. Nitrate levels decreased significantly both in the ischemia (p < 0.01) and ischemia/octreotide (p < 0.01) groups versus control. In the ischemia group, retinal histopathological changes, which were different from the control group, were prominent edema, polymorphonucleated leukocytes infiltration and vacuolated spaces in the inner retina. No significant change was observed in the histopathological specimens of the ischemia/octreotide group. Significant increase in the thickness of the inner plexiform layer of the retina of the ischemia group was observed versus the control and ischemia/octreotide groups (p < 0.01 and p < 0.01, respectively). The thickness of the inner plexiform layer of the retina of the ischemia/octreotide group did not change versus the control group. It was concluded that nitric oxide activity decreased during retinal ischemia-reperfusion and, although octreotide prevented the histopathological damage, it could not ameliorate the nitric oxide activity of the retina. PMID- 12539859 TI - Indirect exposure assessment at the United States Environmental Protection Agency. AB - In the early 1980s, exposures and subsequent health impact assessments from contaminants emitted into the air from stationary sources focused on the inhalation pathway. This 'direct' pathway of exposure was thought to be the most critical pathway, as it is for many contaminants. However, by the latter 1980s, the focus at the Environmental Protection Agency (EPA) shifted to contaminants that would persist in the environment and could bioaccumulate up the food chain. Consumption of impacted food products, and other 'indirect' pathways of exposure, such as soil-related exposures, were shown to result in exposures that exceeded inhalation exposures by two to up to four orders of magnitude. A historical background of indirect exposure assessment at EPA is provided in this paper, followed by an overview of modeling methodologies commonly used in indirect exposure assessments. These methodologies are demonstrated on a contaminant of primary focus for indirect exposure impact, dioxins. Two examples are provided. In the first, an air-to-beef model validation exercise is described. In the second, an indirect exposure assessment on a municipal solid waste incinerator in Ohio conducted by the EPA is summarized. This incinerator emitted very large amounts of dioxin, more than any single source known to EPA. Models were used to predict the movement of dioxins from the stack to a nearby hypothetical farm, where individuals in the farm family were exposed to dioxins from consumption of home-produced beef and milk. The predicted lifetime cancer risk of 2.8 x 10(-4) based on these food pathways was used by EPA's Region 5 to initiate regulatory activity on this incinerator. PMID- 12539858 TI - Spectral characteristics of light sources for S-cone stimulation. AB - PURPOSE: Electrophysiological investigations of the short-wavelength sensitive pathway of the human eye require the use of a suitable light source as a S-cone stimulator. Different light sources with their spectral distribution properties were investigated and compared with the ideal S-cone stimulator. METHODS: First, the theoretical background of the calculation of relative cone energy absorption from the spectral distribution function of the light source is summarized. From the results of the calculation, the photometric properties of the ideal S-cone stimulator will be derived. The calculation procedure was applied to virtual light sources (computer generated spectral distribution functions with different medium wavelengths and spectrum widths) and to real light sources (blue and green light emitting diodes, blue phosphor of CRT-monitor, multimedia projector, LCD monitor and notebook display). The calculated relative cone absorbencies are compared to the conditions of an ideal S-cone stimulator. RESULTS: Monochromatic light sources with wavelengths of less than 456 nm are close to the conditions of an ideal S-cone stimulator. Spectrum widths up to 21 nm do not affect the S-cone activation significantly (S-cone activation change < 0.2%). Blue light emitting diodes with peak wavelength at 448 nm and spectrum bandwidth of 25 nm are very useful for S-cone stimulation (S-cone activation approximately 95%). A suitable display for S-cone stimulation is the Trinitron computer monitor (S-cone activation approximately 87%). The multimedia projector has a S-cone activation up to 91%, but their spectral distribution properties depends on the selected intensity. LCD monitor and notebook displays have a lower S-cone activation (< or = 74%). CONCLUSION: Carefully selecting the blue light source for S-cone stimulation can reduce the unwanted L-and M-cone activation down to 4% for M cones and 1.5% for L-cones. PMID- 12539860 TI - Dietary exposures to persistent organic pollutants. AB - As one of the main components of risk assessment, exposure assessment plays a key role in evaluating risk. Many different scenarios can be developed to estimate the risk from exposure to chemicals such as persistent organic pollutants (POPs). The US Food and Drug Administration (FDA)'s Center for Food Safety and Applied Nutrition (CFSAN) is primarily interested in POPs as humans may be exposed to these compounds through food. Examples of POPs found in food include dioxins, polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers, and some pesticide chemicals. This overview discusses various sources of data that CFSAN has used to estimate dietary exposure to POPs, and provides an example of a recent calculation of an estimate for dietary exposure for consumers in the USA to dioxins in the food supply. PMID- 12539862 TI - Methods for projecting long-term dietary exposure from short-term survey data for environmental contaminants. AB - Public health risk assessments often involve dietary exposures over long periods of time. However, most information about dietary consumption habits comes from short-term surveys that are conducted for periods of three days or less. When employed for characterizing long-term exposures, short-term surveys are likely to underestimate the number of persons consuming a particular food, while overestimating the amount consumed by each individual. Direct application of short-term data is particularly misleading for foods that are consumed infrequently. If a more accurate population estimate for chronic dietary intake is needed for a risk assessment, then two general techniques may be considered. The first method is simpler, while the second is more accurate. Both methods require information about the size of the population consuming the food over the long-term period. The simpler fractional adjustment method reduces consumption across the entire distribution by the ratio of consumer population sizes. Since this method will tend to underestimate high-end exposures and overestimate low end exposures, it is most useful as a quick bounding exercise. Since short-term surveys are better at characterizing the behavior of frequent consumers, a second method employs an exponential function to reduce the low end of the population distribution by a greater amount than the high end. If available, additional information may be used to select the parameter values for the exponential adjustment. Otherwise, an uncertainty range may be used for the parameter values. Since the frequency-based method is more complex, it is most valuable when used as part of a chronic exposure simulation. Examples of both methods are given for the estimation of chronic wine consumption. PMID- 12539861 TI - Travis and Arms revisited: a second look at a widely used bioconcentration algorithm. AB - In 1988, Travis and Arms reviewed the literature and collected data to develop a relationship between the octanol-water partition coefficient (Kow) and the uptake of organic compounds into milk and beef (Travis and Arms, 1988). These equations have been utilized for predicting biotransfer factors for organic chemicals when empirical data are lacking. During the external peer review of the draft US Environmental Protection Agency (EPA) guidance entitled Human Health Risk Assessment Protocol for Hazardous Waste Combustion Facilities (US EPA, 1998) and the development of Superfund's Ecological Soil Screening Levels (US EPA, 2000b), questions challenging the derivation and use of these equations were raised. The primary questions raised were: 1) Are the equations presented in Travis and Arms (1988) for the estimation of transfer of organic compounds from contaminated feed to beef and milk technically valid and reproducible? If so, (2) are the equations appropriate across the entire log Kow range? For these reasons, this study was undertaken to validate the original Travis and Arms equations, to review more recent literature, and. if appropriate, to add to the original Travis and Arms data set to obtain updated equations. This paper presents an evaluation of the original Travis and Arms equations, limitations to their use, and steps to reduce uncertainties associated with their use by updating with more current literature. PMID- 12539863 TI - A review of the effects of uranium and depleted uranium exposure on reproduction and fetal development. AB - Depleted uranium (DU) is used in armor-penetrating munitions, military vehicle armor, and aircraft, ship and missile counterweighting/ballasting, as well as in a number of other military and commercial applications. Recent combat applications of DU alloy [i.e., Persian Gulf War (PGW) and Kosovo peacekeeping objective] resulted in human acute exposure to DU dust, vapor or aerosol, as well as chronic exposure from tissue embedding of DU shrapnel fragments. DU alloy is 99.8% 238Uranium, and emits approximately 60% of the alpha, beta, and gamma radiation found in natural uranium (4.05 x 10(-7) Ci/g DU alloy). DU is a heavy metal that is 160% more dense than lead and can remain within the body for many years and slowly solubilize. High levels of urinary uranium have been measured in PGW veterans 10 years after exposure to DU fragments and vapors. In rats, there is strong evidence of DU accumulation in tissues including testes, bone, kidneys, and brain. In vitro tests indicate that DU alloy may be both genotoxic and mutagenic, whereas a recent in vivo study suggests that tissue-embedded DU alloy may be carcinogenic in rats. There is limited available data for reproductive and teratological deficits from exposure to uranium per se, typically from oral, respiratory, or dermal exposure routes. Alternatively, there is no data available on the reproductive effects of DU embedded. This paper reviews published studies of reproductive toxicity in humans and animals from uranium or DU exposure, and discusses ongoing animal research to evaluate reproductive effects in male and female rats embedded with DU fragments, and possible consequences in F1 and F2 generations. PMID- 12539864 TI - Evaluation of immunotoxicity induced by single or concurrent exposure to N,N diethyl-m-toluamide (DEET), pyridostigmine bromide (PYR), and JP-8 jet fuel. AB - Approximately 5,000 to 80,000 of the US service personnel involved in the Persian Gulf War have complained of a variety of nonspecific symptoms since their return in 1991. These symptoms have been collectively labeled Gulf War Illness and include muscle fatigue, general malaise, myalgia, impaired cognition, ataxia, headaches, fever, joint pain, skin rash, gastrointestinal disturbances, sleep disturbances, and respiratory difficulties. Exposures of military and service personnel were diverse and included the prescribed anti-nerve gas agent pyridostigmine bromide (PYR), N.N-diethyl-m-toluamide (DEET) insect repellent, and environmental exposures to jet fuel. Thus, studies in our laboratory were undertaken to determine if concurrent exposure to these agents, singly or in combination, would contribute to significant alterations in immunological function and disease susceptibility. To assess immune status, eight-week old B6C3F1 female mice were exposed for 14 days to single compounds or tertiary mixtures of 15.5 mg/kg DEET, 2 mg/kg PYR, and 500 mg/kg JP-8 (termed low dose), or 31 mg/kg DEET, 5 mg/kg PYR, and 1,000 mg/kg JP-8 (termed high dose). Immunosuppression was assessed 24 h after the last exposure. No remarkable alterations were evident in hematological parameters, spleen and thymus organ weight and total cellularity, natural killer (NK) cell activity, cytotoxic T-cell activity, or mitogen-induced lymphocyte proliferation after exposure to either single or tertiary mixtures at low or high doses. A few changes in CD4/CD8 flow cytometric lymphocyte subpopulations were detected after exposure to the tertiary mixture at the high dose. Delayed type hypersensitivity (DTH) was decreased by 88% after exposure to the high-dose mixture, and suppression of antibody-specific IgM immune responses (plaque-forming cell, PFC) occurred after exposure to all single and tertiary mixtures at both dose levels. In the PFC response, antagonism was apparent in the mixture, while coexposure to these agents resulted in a synergistic effect in the DTH response. Susceptibility to B16F10 tumor or Listeria monocytogenes challenge was not affected after single or tertiary exposures. These data suggest that combined exposure to DEET, PYR, and JP-8 does not profoundly alter many immunological endpoints, but does selectively target functional endpoints such as the PFC and DTH response. This should be considered when assessing human health risks in the military environment. PMID- 12539865 TI - The use of physiologically based models to integrate diverse data sets and reduce uncertainty in the prediction of perchlorate and iodide kinetics across life stages and species. AB - The effects of perchlorate on the incorporation of iodide into thyroid hormones have been studied for more than 40 years in many species and under varying exposure conditions. Nevertheless, the database for this drinking water contaminant is still incomplete, particularly with regard to human developmental risk. A method for integrating the available data and forming meaningful conclusions for risk assessment is needed. To this end, an initial suite of physiologically based pharmacokinetic (PBPK) models has been developed, which incorporates physiological data for the relevant species and life stages and kinetic data for perchlorate and iodide, as well as the interaction between the two anions. The validated models successfully describe perchlorate-induced inhibition of thyroid iodide uptake and perchlorate and iodide kinetics in the male, pregnant, lactating, fetal, and neonatal rats and the adult humans. The relationships of model-predicted internal dose metrics and kinetic parameters allow a direct comparison of internal dose metrics across life stages in rats and humans. By incorporating all the available data, these models provide a framework for species and life stage extrapolation where the lack of specific data sets would otherwise limit predictive capability. This paper demonstrates two approaches for calculating life stage-specific equivalent doses in a risk assessment for perchlorate: the direct combination of validated model predictions, and the development of preliminary PBPK models for the human sensitive populations based on the relationship of the parameters in the validated rat and human models. Either approach can be used to perform the needed dosimetry. However, the second approach provides the advantage of a preliminary human life stage-specific PBPK model that can be used for identification of key data gaps and estimation of uncertainty. PMID- 12539866 TI - Defining populations: a key step in identifying spatial and temporal scales. AB - The scale at which ecological risk assessors frame their analyses can be driven by a number of considerations. In many cases, the selection of the scale is among the most important decisions within the 'problem-formulation' phase of the ecological risk characterization. The scale of a risk characterization is often defined based on ownership boundaries, ecological, or habitat boundaries and/or physical and hydrogeological boundaries (e.g., watersheds). With increasing value being placed on consideration of spatial scales in risk characterization, the need to define a scale that balances analytical and ecological realities is imperative. Defining the spatial scale based on the scale of a population offers the risk assessor the opportunity to improve the ecological relevance of the risk characterization. Arriving at a single operational definition of a population would be difficult and not appropriate because of the variability in sites, issues, and management goals; however, we can begin to review operational definitions and establish analytical frameworks by which to select the appropriate, site-specific scale within the problem-formulation phase. Defining a scale based on a combination of ecological and site ownership considerations and the inclusion of habitat quality in the assessment of scale can improve the power and relevance of the problem formulation and risk characterization. In this analysis, we assess methods for incorporating spatial considerations into risk analyses, beginning with developing an understanding of the context of a site within the larger landscape. Clearly defining the scale of an assessment including the operational definition of populations is an important step in the problem formulation. Within a site or landscape, the scale may be refined further based on the percentage of the land within the scale of the analysis that provides habitat for a given species. PMID- 12539867 TI - Spatially explicit exposure models: application to military sites. AB - As a result of defense-related activities, large areas of valuable habitats can be physically disturbed and/or contaminated by hazardous pollutants. It is, however, important to understand that the military impact on such areas may not be wholly detrimental to the environment: their closure to the public can result in environmental conservation of large portions of habitat. Many former military ranges have been converted for civil usage; others are still actively operated for military purposes. In both cases, a major task for environmental policy- and decision-makers is to determine scientifically justified reuse and/or operation scenarios that would not result in deterioration of site habitats and that would in fact be favorable to further development of biodiversity within the areas' existing ecosystems. Ecological risk assessment is suggested as the key process to use in facilitating the assessment of the ecological value of contaminated and/or disturbed military sites and in the development of a reuse decision protocol. We are currently developing and integrating a number of risk and habitat assessment techniques into such a protocol. This paper presents our approach to one aspect of this system: the incorporation of spatial information into ecological risk assessment. We present a software prototype that calculates radionuclide accumulation by deer foraging in areas having specified contamination patterns. PMID- 12539868 TI - Using landscape ecology to focus ecological risk assessment and guide risk management decision-making. AB - Ecological risk assessment (EcoRA) generally suffers from limited application of ecological knowledge in the definition and characterization of real-world sites. Not surprisingly, most remediation decisions, which follow, have little or no relationship to the valued ecological resources of the site or the broader region. The practice has evolved to favor engineering-based mitigation strategies, which eliminate excess chemical concentrations at sites, or otherwise break exposure pathways, but which may not be ecologically beneficial. The heavy emphasis of EcoRA on toxicity threshold levels tends to focus dollars on clean up of small areas or volumes with high concentrations. Moreover, intrusive remediation technologies often render an area uninhabitable to the very species that were to be protected. Infusion of ecological knowledge into EcoRA has been difficult. Most professional ecologists choose not to venture into the messy applied fields, leaving their impressive knowledge untapped. Moreover, narrowly defined responsibilities within government circles can limit cooperation and coordination. The realization that land use activities often have greater adverse consequences to wildlife than do chemicals provides an opportunity to change attitudes and practices. We are developing procedures that incorporate landscape features into the environmental management process. Specifically, we are using an iterative approach to: a) identify scenarios where habitat value is important in EcoRAs; b) guide selection of appropriate assessment species, i) keyed to wildlife distribution ranges; ii) keyed to a database of habitat suitability models; iii) cross-linked with the EPA exposure handbook species; iv) referenced to wildlife distributions (e.g., breeding bird survey); c) define data collection needs for reconnaissance-, screening-, and definitive-level characterization of habitat quality for potential assessment species; d) generate spatially explicit descriptions of habitat quality for various assessment species; and e) allocate exposure estimates using both habitat quality and spatial variations in chemical concentration. These refinements in theEcoRA process are expected to improve risk estimates and provide valuable information to be used instructuring risk management options. The approach can guide the planning process so that an assessment considers the most relevant species of the area and defines the relevant parameters to be measured. In risk characterization, these data are used to calculate more realistic exposure assessments. In guiding remediation, the approach logically considers a wider range of land management options than are considered at most sites today. For example, habitat enhancement can be used to draw animals away from contaminated zones. Contaminated localities that also have poor-quality habitat may be allowed to go through a slower, less costly bioremediation process until the risk level is lowered to acceptable levels. And direct comparisons of lost resources stemming from destructive remediation options can be assessed instead of merely focusing on the lowering of contaminant concentrations. This paper presents the conceptual foundation for incorporating landscape ecology into the risk assessment process. PMID- 12539869 TI - Occupational exposures during the World Trade Center disaster response. AB - Upon the request of the New York City Department of Health, the Centers for Disease Control and Prevention's National Institute for Occupational Safety and Health (NIOSH) monitored occupational exposures among emergency response workers during the rescue and recovery activities at the World Trade Center disaster site from September 18 through 4 October 2001. During this period, over 1,200 bulk and air samples were collected to estimate or characterize workers' occupational exposures. Samples were collected and analyzed for asbestos, carbon monoxide (CO), chlorodifluoromethane (Freon 22), diesel exhaust, hydrogen sulfide, inorganic acids, mercury and other metals, polynuclear aromatic hydrocarbons, respirable particulate not otherwise regulated (PNOR), respirable crystalline silica, total PNOR, and volatile organic compounds. Exposures to most of these potential hazards did not exceed NIOSH Recommended Exposure Limits or Occupational Safety and Health Administration Permissible Exposure Limits. However, one torch cutter was overexposed to cadmium and another worker (and possibly three others) was overexposed to CO. The elevated cadmium and CO levels were the result of workers using oxy-acetylene cutting torches and gasoline powered cutting saws. Recommendations were made to ensure adequate ventilation and worker understanding when using these tools and, where possible, to substitute rechargeable, battery-powered cutting saws for gasoline-powered ones. Toxicology PMID- 12539870 TI - Pentagon surface wipe sampling health risk assessment. AB - Within hours after the terrorist attack on the Pentagon on September 11, 2001, an extensive sampling effort was initiated within the building to include surface wipe sampling for select coplanar polychlorinated biphenyls (PCBs), congeners of polychlorinated dibenzo(p)dioxins and dibenzofurans (dioxins/furans) and lead. A risk-based screening level method was utilized to determine the necessity of additional sampling and to assess the potential for emergency response crews, remediation crews, and returning Pentagon workers to have adverse health impact from exposure to the PCBs. dioxins/furans, and lead on nonporous surfaces. The screening level method included all exposure pathways: dermal absorption, incidental ingestion, and inhalation. Various U.S. Environmental Protection Agency (USEPA) guidance documents provided the basis for the method. The underlying assumptions were that the PCBs, dioxins/furans, and lead were contained in the dust or soot layer found on nonporous surfaces, that sampling results were representative of the surfaces from which they were taken, and that the analytical methods employed were able to detect 100% of these substances. A few of the limitations associated with this method included the lack of toxicity values for dermal absorption and the lack of accurate, discrete dermal-exposure values. Evaluation results indicated that additional sampling was not necessary and that concentrations of PCBs, dioxins/furans, and lead on nonporous surfaces were below levels expected to cause adverse health impacts to emergency response crews, remediation crews, and returning Pentagon workers. PMID- 12539871 TI - Identification of target genes responsive to JP-8 exposure in the rat central nervous system. AB - Concern for the health risk associated with occupational exposure to jet fuel has emerged in the Department of Defense. Jet propulsion fuel-8 (JP-8) is the fuel used in most US and North Atlantic Treaty Organization (NATO) jet aircraft, and will be the predominant fuel both for military land vehicles and aircraft into the twenty-first century. JP-8 exhibits reduced volatility and lower benzene content as compared to JP-4, the predominant military aircraft fuel before 1992, possibly suggesting greater occupational exposure safety. However, the higher rates of occupational exposure through fueling and maintenance of increasingly larger numbers of aircraft/vehicles raise concerns with respect to toxicity. Clinical studies of workers experiencing long-term exposure to certain jet fuels demonstrated deficits in CNS function, including fatigue, neurobehavioral changes, psychiatric disorders, and abnormal electroencephalogram (EEG). In the present study, cDNA nylon arrays (Atlas Rat 1.2 Array, Clontech Laboratories, Palo Alto, CA) were utilized to measure changes in gene expression in whole brain tissue of rats exposed repeatedly to JP-8, under conditions that simulated possible real-world occupational exposure (6 h/day for 91 days) to JP-8 vapor at 1,000 mg/m3. Gene expression analysis of the exposure group compared to the control group revealed a modulation of several genes, including glutathione S transferase Yb2 subunit (GST Yb2); cytochrome P450 IIIAl (CYP3A1); glucose dependent insulinotropic peptide (GIP); alpha1-proteinase inhibitor (alpha1-AT); polyubiquitin; GABA transporter 3 (GAT-3); and plasma membrane Ca2+-transporting ATPase (brain isoform 2) (PMCA2). The implications of these vapor-induced changes in gene expression are discussed. PMID- 12539872 TI - Potential technology for studying dosimetry and response to airborne chemical and biological pollutants. AB - Advances in computational, and imaging techniques have enabled the rapid development of three-dimensional (3-D) models of biological systems in unprecedented detail. Using these advances, 3-D models of the lungs and nasal passages of the rat and human are being developed to ultimately improve predictions of airborne pollutant dosimetry. Techniques for imaging the respiratory tract by magnetic resonance imaging (MRI) were developed to improve the speed and accuracy of geometric data collection for mesh reconstruction. The MRI resolution is comparable to that obtained by manual measurements but at much greater speed and accuracy. Newly developed software (NWGrid) was utilized to translate imaging data from MR into 3-D mesh structures. Together, these approaches significantly reduced the time to develop a 3-D model. This more robust airway structure will ultimately facilitate modeling gas or vapor exchange between the respiratory tract and vasculature as well as enable linkages of dosimetry with cell response models. The 3-D, finite volume, viscoelastic mesh structures form the geometric basis for computational fluid dynamics modeling of inhalation, exhalation and the delivery of individual particles (or concentrations of gas or vapors) to discrete regions of the respiratory tract. The ability of these 3-D models to resolve dosimetry at such a high level of detail will require new techniques to measure regional airflows and particulate deposition for model validation. PMID- 12539873 TI - A typical path model of tracheobronchial clearance of inhaled particles in rats. AB - A mathematical description of particle clearance from the ciliated conducting airways (tracheobronchial region) of the lungs in rats was developed, assuming that particles on the mucus blanket behave as a fluid and adhere to principles of fluid flow described by the continuity equation. Effective particle transport velocities for given generations of airways were estimated from reported tracheal mucus velocities. Using typical rat airway geometry and estimated particle transport velocities, solutions of sets of rate equations for transport from each generation of airways were summed to estimate total particle clearance from the tracheobronchial region of the lung as a function of time. Aerosol particle size distribution (MMAD ranging from 0.1 to 4.2 microm, and sigma(g) from 1 to 2.7) and concentration data from several investigators were used to predict short term, tracheobronchial clearance (retention) in rats up to 24 h following exposure. Comparisons between predicted and observed retention showed an average difference between model predictions, and observed fractional retention of initial lung or body burden was 4.9%, with a tendency toward underprediction of clearance of particles >3.0 microm. PMID- 12539874 TI - An 'injury-time integral' model for extrapolating from acute to chronic effects of phosgene. AB - The present study compares acute and subchronic episodic exposures to phosgene to test the applicability of the 'concentrationxtime' (CxT) product as a measure of exposure dose, and to relate acute toxicity and adaptive responses to chronic toxicity. Rats (male Fischer 344) were exposed (six hours/day) to air or 0.1, 0.2, 0.5 and 1.0 ppm of phosgene one time or on a repeated regimen for up to 12 weeks as follows: 0.1 ppm (five days/week), 0.2 ppm (five days/week), 0.5 ppm (two days/week), or 1.0 ppm (one day/week) (note that the CxT for the three highest exposures was the same). Animals were sacrificed at 4, 8, and 12 weeks during the exposure and after four weeks recovery. Bronchoalveolar lavage (BAL) was performed 18 hours after the last exposure for each time period and the BAL supernatant assayed for protein. Elevated BAL fluid protein was defined as 'acute injury', diminished response after repeated exposure was defined as 'adaptation', and increased lung hydroxyproline or trichrome staining for collagen was defined as 'chronic injury'. Results indicated that exposures that cause maximal chronic injury involve high exposure concentrations and longer times between exposures, not high CxT products. A conceptual model is presented that explains the lack of CxT correlation by the fact that adaptation reduces an 'injury-time integral' as phosgene exposure is lengthened from acute to subchronic. At high exposure concentrations, the adaptive response appears to be overwhelmed, causing a continued injury-time integral, which appears to be related to appearance of chronic injury. The adaptive response is predicted to disappear if the time between exposures is lengthened, leading to a continued high injury-time integral and chronic injury. It has generally been assumed that long, continuous exposures of rodents is a conservative approach for detecting possible chronic effects. The present study suggests that such an approach my not be conservative, but might actually mask effects that could occur under intermittent exposure conditions. PMID- 12539875 TI - Response of F344 rats to inhalation of subclinical levels of sarin: exploring potential causes of Gulf War illness. AB - Subclinical, repeated exposures of F344 rats to sarin resulted in brain alterations in densities of chlonergic receptor subtypes that may be associated with memory loss and cognitive dysfunction. The exposures also depressed the immune system. The rat appears to be a good model for studying the effects of subclinical exposure to a nerve gas. PMID- 12539876 TI - Communicating risk in diverse communities. AB - Environmental risk communication involves the use of many tools and techniques to form and deliver effective messages. These messages need to inform affected communities about the potential risks and health effects resulting from exposures to environmental contaminants. More importantly, the messages need to reflect the needs, concerns, sensitivities, and knowledge base of the affected community. This risk message is based on profiling, which is simply getting to know your community (demographically, culturally, and historically) and melding this information with scientific knowledge to effectively deliver a message. Three Air Force (AF) case studies are used to discuss how effective risk-communication methods can be utilized in vastly different communities: an on-base residential population, an AF occupational setting, and a remote Alaskan community impacted by historical AF activities. We will discuss the methods used to profile affected communities, determine and deliver messages, and describe the reaction and receptiveness of the community receiving the message. PMID- 12539877 TI - An overview of human exposure modeling activities at the USEPA's National Exposure Research Laboratory. AB - The computational modeling of human exposure to environmental pollutants is one of the primary activities of the US Environmental Protection Agency (USEPA)s National Exposure Research Laboratory (NERL). Assessment of human exposures is a critical part of the overall risk assessment paradigm. In exposure assessment, we analyze the source-to-dose sequence of processes, in which pollutants are released from sources into the environment, where they may move through multiple environmental media, and to human receptors via multiple pathways. Exposure occurs at the environment-human interface, where pollutants are contacted in the course of human activities. Exposure may result in a dose, by which chemicals enter the body through multiple portals of entry, primarily inhalation, ingestion, and dermal absorption. Within the body, absorbed pollutants are distributed to, metabolized within, and eliminated from various organs and tissues, where they may cause toxicologic responses or adverse health effects. The NERL's modeling efforts are directed at improving our understanding of this sequence of processes, by characterizing the various factors influencing exposures and dose, and their associated variabilities and uncertainties. Modeling at the NERL is one of three essential programmatic elements, along with measurements and methods development. These are pursued interactively to advance our understanding of exposure-related processes. Exposure models are developed and run using the best currently available measurement data to simulate and predict population exposure and dose distributions, and to identify the most important factors and their variabilities and uncertainties. This knowledge is then used to guide the development of improved methods and measurements needed to obtain better data to improve the assessment and reduce critical uncertainties. These models and measurement results are tools that can be used in risk assessments and in risk management decisions in order to reduce harmful exposures. Current areas of the NERL's exposure modeling emphasis include: Pollutant concentrations in ambient (outdoor) air using the Third Generation Air Quality Modeling System's Community Multiscale Air Quality model (Models-3/CMAQ); Air flow and pollutant concentrations at local and microenvironmental scales using computational fluid dynamics (CFD); Human inhalation exposure to airborne particulate matter, air toxics, and multipathway exposure to pesticides, using the Stochastic Human Exposure and Dose Simulation (SHEDS) model; Human and ecological exposure and risk assessments of hazardous waste sites using Framework for Risk Analysis in Multimedia Environmental Systems--Multimedia, Multipathway, Multireceptor Risk Assessment (FRAMES-3MRA), one of many software programs available from the NERL's Center for Exposure Assessment Modeling (CEAM); Physiologically based pharmacokinetic (PBPK) modeling of pesticides and volatile organic compounds (VOCs) in the Exposure-Related Dose-Estimating Model (ERDEM). A brief historical overview of the NERL's evolution of human exposure models is presented, with examples of the present state-of-the-science represented by SHEDS and FRAMES-3MRA. PMID- 12539878 TI - Application of in vitro biotransformation data and pharmacokinetic modeling to risk assessment. AB - The adverse biological effects of toxic substances are dependent upon the exposure concentration and the duration of exposure. Pharmacokinetic models can quantitatively relate the external concentration of a toxicant in the environment to the internal dose of the toxicant in the target tissues of an exposed organism. The exposure concentration of a toxic substance is usually not the same as the concentration of the active form of the toxicant that reaches the target tissues following absorption, distribution, and biotransformation of the parent toxicant. Biotransformation modulates the biological activity of chemicals through bioactivation and detoxication pathways. Many toxicants require biotransformation to exert their adverse biological effects. Considerable species differences in biotransformation and other pharmacokinetic processes can make extrapolation of toxicity data from laboratory animals to humans problematic. Additionally, interindividual differences in biotransformation among human populations with diverse genetics and lifestyles can lead to considerable variability in the bioactivation of toxic chemicals. Compartmental pharmacokinetic models of animals and humans are needed to understand the quantitative relationships between chemical exposure and target tissue dose as well as animal to human differences and interindividual differences in human populations. The data-based compartmental pharmacokinetic models widely used in clinical pharmacology have little utility for human health risk assessment because they cannot extrapolate across dose route or species. Physiologically based pharmacokinetic (PBPK) models allow such extrapolations because they are based on anatomy, physiology, and biochemistry. In PBPK models, the compartments represent organs or groups of organs and the flows between compartments are actual blood flows. The concentration of a toxicant in a target tissue is a function of the solubility of the toxicant in blood and tissues (partition coefficients), blood flow into the tissue, metabolism of the toxicant in the tissue, and blood flow out of the tissue. The appropriate degree of biochemical detail can be added to the PBPK models as needed. Comparison of model simulations with experimental data provides a means of hypothesis testing and model refinement. In vitro biotransformation data from studies with isolated liver cells or subcellular fractions from animals or humans can be extrapolated to the intact organism based upon protein content or cell number. In vitro biotransformation studies with human liver preparations can provide quantitative data on human interindividual differences in chemical bioactivation. These in vitro data must be integrated into physiological models to understand the true impact of interindividual differences in chemical biotransformation on the target organ bioactivation of chemical contaminants in air and drinking water. PMID- 12539879 TI - A spatially and bioenergetically explicit terrestrial ecological exposure model. AB - In typical exposure models, dose is a function of ingestion rate, which is a function of field metabolic rate and food energy availability. It is implicitly assumed that neither food energy nor ingestion rate is limited. This is unlikely to be true in the field. Poor habitat quality (expressed as limited or lacking food energy) or a physiologically limited maximum ingestion rate may collectively limit energy intake. A receptor may thus be as much at risk from lack of energy as from toxicant effects. To explore this possibility, an existing spatially explicit exposure model (SE3M) was enhanced to: 1) express 'habitat quality' in terms of gross energy available from a suite of habitat-specific food types, 2) track fulfillment of a receptor's daily energy needs as it traverses habitat patches with varying gross energy levels, 3) link intake of contaminants to food consumed to meet daily energy needs, and 4) track contaminant doses and resulting tissue residue levels as a receptor moves through habitat patches with differing levels of contamination. A feedback term through which chemical stressors affect a receptor's ability to intake and process energy was not considered at this time. The now spatially and energetically explicit exposure model, SE(4)M, provides a platform for exploring spatial and bioenergetic factors that may influence a receptor's acquisition of energy and contaminant tissue residues as it moves through space and time. An application of this model would be to provide predictions of tissue residue levels that are accessible to calibration or validation with empirical field data. PMID- 12539880 TI - Self-prescribed laxative use: a drug-utilization review. AB - This study was conducted to determine the reasons for the choice of self prescribed laxatives and to acquire information on how they were used and tolerated. From November 1999 to February 2000, 70 pharmacies, uniformly located throughout the Campania region of southern Italy, distributed a questionnaire to purchasers of over-the-counter laxatives. The average age of the (mostly female) respondents was 45.9 years; 23.8% were elderly. Among the 7324 individuals who completed the survey, 77.6% selected an oral product; 22.4% preferred rectal administration. A physician influenced the choice of a laxative in 37.7% of the cases, a pharmacist in 20.5%; other suggestions came from relatives (14%), acquaintances (12.1%), advertisements (11.7%), and miscellaneous sources (4%). Only 59.8% of respondents used these drugs correctly, and 58.2% consulted a physician or pharmacist because of constipation. Adverse effects, mainly gastrointestinal symptoms, occurred in 6.1% of those surveyed. The long-term use or abuse of laxatives can cause serious medical consequences, as well as mask diseases, delaying diagnosis and appropriate treatment. Physicians, pharmacists, and other health-care personnel should counsel patients on the proper use of these easily available, ubiquitous drugs. PMID- 12539881 TI - Arnica montana gel in osteoarthritis of the knee: an open, multicenter clinical trial. AB - This open multicenter trial investigated the safety and efficacy of an Arnica montana fresh plant gel, applied twice daily, in 26 men and 53 women with mild to moderate osteoarthritis (OA) of the knee. After 3 and 6 weeks, significant decreases in median total scores on the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) were evident in the intention-to-treat and per protocol populations (both P < .0001). Scores on the pain, stiffness, and function subscales also showed significant reductions at these timepoints. The overall local adverse-event rate of 7.6% included only one allergic reaction. Sixty-nine patients (87%) rated the tolerability of the gel as "good" or "fairly good," and 76% would use it again. Topical application of Arnica montana gel for 6 weeks was a safe, well-tolerated, and effective treatment of mild to moderate OA of the knee. PMID- 12539882 TI - Spermicidal activity of a new contraceptive sponge. AB - An open, crossover study evaluated the spermicidal efficacy and irritative potential of the Protectaid sponge. Postcoital tests were conducted in nine healthy couples under control conditions (no sponge) and with the sponge. Microscopic examination of postcoital mucus from the female partners showed that sperm entered the cervix in all nine controls, whereas entry was completely prevented by the sponge in four of nine women. None of the participants showed colposcopic evidence of mucosal irritation while using the sponge. Colposcopic abnormalities were evident in two of nine women during control testing. The urgent need for female-controlled contraceptive methods with the potential to protect against sexually transmitted diseases makes this low-dose spermicidal vaginal sponge an effective and safe alternative. PMID- 12539883 TI - IV-to-oral switch therapy for community-acquired pneumonia requiring hospitalization: focus on gatifloxacin. AB - The majority of the 1.1 million patients hospitalized for community-acquired pneumonia (CAP) in the United States begin therapy with an intravenous antibiotic. A switch to oral therapy as soon as patients are clinically stable reduces the length of hospitalization and associated costs. Fluoroquinolones are appropriate candidates for switch therapy. Gatifloxacin is an excellent choice when a fluoroquinolone is being considered for sequential switch therapy in the treatment of CAP requiring hospitalization. PMID- 12539885 TI - I resolve to... PMID- 12539884 TI - Charcoal tablets in the treatment of patients with irritable bowel syndrome. AB - This double-blind, randomized, multicenter, prospective clinical trial evaluated a commercial formulation of charcoal tablets (Eucarbon) and tablets containing only nonactivated charcoal (carbo ligni [CL]) in 284 patients between the ages of 19 and 70 years with irritable bowel syndrome (IBS). After 12 weeks, 262 patients were available for intention-to-treat analysis. Overall well-being, the primary efficacy parameter, was determined by means of the Francis scoring system. Eucarbon treatment alleviated symptoms by about 60%, but the relative gain in efficacy vis-a-vis the CL group was only 9%. Several clinical observations and subgroup analyses, however (eg, in patients suffering from constipation), showed that Eucarbon was more effective. Both treatments were well tolerated and produced adverse events at a similar frequency (22%, Eucarbon vs 17%, CL). In most cases, it was not possible to distinguish the adverse event from symptoms of IBS. The herbal preparation Eucarbon was effective and safe in IBS, a complex disease requiring long-term treatment. PMID- 12539886 TI - Driving with a visual impairment: new guidelines are needed. PMID- 12539887 TI - UV protection. PMID- 12539888 TI - Who is the para? PMID- 12539889 TI - Awareness of age-related macular degeneration in adults: the results of a large scale international survey. AB - BACKGROUND: Early detection and prompt implementation of appropriate rehabilitation services and therapy can help minimize the long-term disability that results from age-related macular degeneration (ARMD). The level of awareness of ARMD among the general public may influence early detection of this disease. METHODS: Random samples of European and North American populations were interviewed in an international survey designed to assess the attitudes of people to eye examinations, awareness of the causes of vision loss, awareness of ARMD, and knowledge of its treatment options. RESULTS: A total of 7,830 adults of at least 18 years of age were questioned. The main finding was that knowledge of ARMD is poor-the majority of respondents (70%) were not at all familiar with ARMD. Only 2% of participants knew that ARMD is the leading cause of severe vision loss in adults 50 years of age or older, while a majority of those surveyed (75%) was unaware of treatments available for ARMD. CONCLUSIONS: These data show that there is low public awareness of ARMD, which may result in people with ARMD not receiving prompt medical advice and treatment. There is a need for a wide-ranging educational initiative to increase awareness of ARMD and highlight the importance of undergoing regular eye examinations. PMID- 12539890 TI - Association of symptoms and convergence and accommodative insufficiency in school age children. AB - BACKGROUND: The purpose of this article was to investigate the association between convergence insufficiency (CI) and accommodative insufficiency (AI) and symptoms in a group of school-aged children. METHODS: Children ages 8 to 15 years were recruited from two public and 2 private elementary schools in Southern California. The CI Symptom Survey (CISS) was administered to all children before a Modified Clinical Technique vision screening. Children with normal visual acuity, minimal uncorrected refractive error, and no strabismus were tested for CI and Al. RESULTS: Four hundred sixty nine children were initially screened and 392 participated in testing for CI and AI. Fifty-five percent of the children (218) were classified as having normal binocular vision (NBV), 4.6% (18) had three signs of CI, 12.7% (50) had two signs of CI, 10.5% (41) were classified as AI (with no signs of CI), and 16.6% (65) were classified as other. The symptom score was 3.78 for the NBV group, 4.6 for the two-sign CI group, 6.67 for the three-sign CI group, and 6.37 for the Al group. The three-sign CI and the Al groups scored significantly higher than the NBV group on the CISS (p < or = 0.001). CONCLUSION: CI and AI are common conditions in school-age children and are associated with increased symptoms. PMID- 12539891 TI - Visual factors that significantly impact academic performance. AB - BACKGROUND: Both race and socio-economic status are correlated to performance in the classroom. These two factors are inter-related, since minorities, proportion wise, are more highly represented in the lower socio-economic strata. Inefficient visual skills have been shown to be more prevalent among minority groups and in low socio-economic groups. These inefficient visual skills impact the students' learning. This study was undertaken to discover the visual skills that were significantly correlated with academic performance problems. METHOD: A total of 2,659 examinations were performed on 540 children over the course of six examination periods, which were administered over three consecutive school years. Socio-economic, racial, and standardized academic performance data (Iowa Test of Basic Skills--ITBS) were furnished by the families and the school system. The visual and demographic data from the examinations were then compared to performance on the 21 subtests of the ITBS. RESULTS: Some visual factors were found to be a much better predictor of scores on the ITBS than either race or socio-economic status. Even though the significance of these two demographic variables was small, race and socio-economic variables were each significant in about a third of the 21 ITBS scores. CONCLUSION: Visual factors are significantly better predictors of academic success as measured by the ITBS than is race or socio-economics. Visual motor activities are better predictors of ITBS scores than are binocularity or accommodation. These latter skills were significant predictors also, but to a lesser degree. PMID- 12539892 TI - Auditory response to a photic stimulus. AB - BACKGROUND: The photic sneeze response is a well-established clinical phenomenon in which patients respond by sneezing in an uncontrolled manner. This paroxysm of sneezing is evoked by a sudden exposure to an intense or bright light source. The pathogenesis of this response is apparently quite complex. Sudden exposure to bright light involves the optic, oculomotor, and trigeminal nerves, the autonomic nervous system and central brainstem functions. Anecdotal reports have suggested the photic sneezing response may occur as an autosomal dominantly inherited condition. However, such a familial pattern is not always present. METHODS: This article presents a descriptive report of a case in which a patient sought comprehensive eye care and, during the course of the examination, reported a relationship between light and a predictable auditory response. The mechanism for this involuntary auditory response to a bright light may be similar to that involved with photic sneeze response. RESULTS: This patient's history, specific features of the auditory response, and ophthalmic findings as a result of the comprehensive eye examination are reported. CONCLUSION: The photic sneeze response is not an uncommon phenomenon. A similar mechanism may explain the auditory response to a photic stimulus reported in this article. PMID- 12539893 TI - Vague persistent patient complaints. PMID- 12539894 TI - Accrual vs. cash accounting. PMID- 12539895 TI - Popular Paraoptometric Certification program expands examination schedule. PMID- 12539896 TI - The insanity defense and game theory: reflections on Texas v. Yates. PMID- 12539897 TI - The insanity defense, continued. PMID- 12539898 TI - The MacArthur Competence Assessment Tool-Fitness to Plead: a preliminary evaluation of a research instrument for assessing fitness to plead in England and Wales. AB - This study concerns the preliminary evaluation of an instrument, the MacArthur Competence Assessment Tool-Fitness to Plead (MacCAT-FP), for assessing competence to stand trial, according to the legal requirements in England and Wales. The purposes of the study were to ascertain whether the instrument can be administered practically to groups of prisoners, both those with mental illness and those without; to examine its internal consistency and interrater reliability; to determine whether it can distinguish between fit and unfit individuals, as judged by expert forensic psychiatrists; and to discover whether it is sensitive to changes over time. The instrument was administered to two groups of remanded prisoners: those transferred to psychiatric units for treatment and those without mental illness. In addition to the MacCAT-FP, scales measuring symptom severity and IQ were administered to all participants. Results suggest that the instrument is practical, with good internal consistency and interrater reliability. The correlation of scores with psychiatrists' opinions as to fitness was 0.77. Scores of unfit patients were significantly different from those of fit individuals. The instrument was able to detect significant differences in scores over time. It is suggested that once the instrument has been further validated for research purposes, it may be developed for clinical application. PMID- 12539899 TI - Commentary: Incorporation of competence instruments into clinical practice. PMID- 12539900 TI - Commentary: Competence assessment practices in England and Australia versus the United States. PMID- 12539901 TI - The role of mental health in the inmate disciplinary process: a national survey. AB - An effective system of inmate discipline is an important aspect of a safely run prison or jail. Historically, mentally ill inmates have had few or no protections against discipline routinely applied to their non-mentally ill peers. Arising from recent class action lawsuits challenging the quality of mental health care delivery in the nation's prisons, prison mental health professionals have been called on to play an increasing role in the inmate disciplinary process. Referral questions include whether an inmate is competent to proceed with disciplinary proceedings and whether mental illness may have contributed to the rule violation. Prison mental health professionals participating in inmate disciplinary proceedings must therefore be familiar with relevant clinical, legal, and ethics issues. Little has been written in the psychiatric literature, however, examining this important role for prison mental health professionals. After first reviewing core legal and constitutional concepts, the author presents the results of a nationwide survey examining the role for mental health professionals in the inmate disciplinary process. To the author's knowledge, this is the first study to provide a comprehensive review of this subject. PMID- 12539902 TI - Commentary: The role of mental health in the inmate disciplinary process. PMID- 12539903 TI - Commentary: Mental health in the inmate disciplinary process. PMID- 12539904 TI - Personality disorders and violence among female prison inmates. AB - The current study seeks to expand our understanding of the increasingly well documented relationship between mental disorder and violence, specifically by examining the relationship between Axis II disorders and community and institutional violence among a cohort of 261 incarcerated women. Drawing from an initial screening of 802 female inmates in maximum security, we sampled to identify 200 nonpsychotic women who met criteria for one of the four Cluster B personality disorders, and 50 nonpsychotic women who did not meet criteria for these disorders. Each inmate was interviewed with the Structured Clinical Interview for DSM-IV Axis II Personality Disorders (SCID-II). Information regarding instant offense and institutional behavior was obtained from prison files and a self-report inventory. The analyses indicated a high degree of comorbidity between the various Cluster B diagnoses and a significant association with various types of violent crime and nonviolent criminality. Significant relationships were found between Antisocial Personality Disorder and institutional violence, and Narcissistic Personality Disorder and incarceration for a violent crime. Cluster A diagnosis was unexpectedly found to be associated with both incarceration for a violent crime and incarceration for prostitution. PMID- 12539905 TI - Commentary: The forensic relevance of personality disorder. PMID- 12539906 TI - A comparison of substance abuse among female offender subtypes. AB - The relationship between substance abuse and female criminal offending has been understudied. To aid in clarification of this relation, substance use histories of female offender subtypes were compared. Participants were 152 female prison inmates subgrouped on the basis of offense category: violent (n = 79), property (n = 39), and drug (n = 34). As hypothesized, substance use histories differed across offender subgroups. Violent offenders were most clearly distinguished from other offender subgroups. The data provide evidence linking alcohol, combined cocaine and alcohol, and marijuana misuse with serious violent offending among women and show that violent offenders, compared with other offender subgroups, perceive less association between alcohol consumption and alcohol-related negative consequences. PMID- 12539907 TI - Novel cases: malingering by animal proxy. AB - Malingering to obtain medications of abuse is well documented in the general medical setting. However, we have found no cases previously reported of such malingering in a veterinary setting. We report five cases submitted by veterinarians in which clients (pet owners) are strongly suspected or confirmed to have been engaging in malingering to obtain controlled medications for their personal use. Cases bear a striking resemblance to malingering in the general medical setting for drugs to abuse. We propose that veterinarians, like their medical counterparts, are potential targets of malingering by their clients for drugs of abuse. Because of their familiarity with this condition, psychiatrists may have a role in training veterinarians to recognize malingering on the part of their human clients. In addition, psychiatrists may benefit from familiarizing themselves with novel forms of malingering, such as are presented in this case series. PMID- 12539908 TI - The civil commitment of sexually violent predators: a unique Texas approach. AB - Numerous states have enacted statutes focusing on the civil commitment of sexually violent predators. The Texas statute, like many others, calls for the involuntary commitment of those with a mental abnormality--specifically in Texas, a behavioral abnormality--who are likely to engage in predatory acts of sexual violence. All of these states, except Texas, have passed legislation creating inpatient treatment for those persons committed. Texas, instead, has focused exclusively on the use of outpatient treatment and supervision as an alternative to the route of inpatient commitment This article is focused on the development and implementation of the Texas statute with an emphasis on forensic assessment, expert testimony, and risk assessment. PMID- 12539909 TI - Knowledge is not power--knowledge is obligation. PMID- 12539910 TI - A comparison of R. v. Stone with R. v. Parks: two cases of automatism. PMID- 12539911 TI - A call for more program evaluation of forensic outpatient clinics: the need to improve effectiveness. PMID- 12539912 TI - Punishing the insane: the verdict of guilty but mentally ill. PMID- 12539913 TI - Commentary: Punishing the unpunishable--the abuse of psychiatry to confine those we love to hate. PMID- 12539914 TI - Commitment versus confinement: therapeutic passes in the management of insanity acquittees. PMID- 12539915 TI - What price security?--A review of Steven Spielberg's Minority report. PMID- 12539916 TI - Countertransference and its relationship with forensic psychiatry. PMID- 12539917 TI - Response to Munro: "Judicial psychiatry in China and its political abuses". PMID- 12539918 TI - "Killing" (Nadelson T: J Am Acad Psychiatry Law 30:201-6, 2002). PMID- 12539919 TI - What washing elephants can teach us about implementing changes to practice. PMID- 12539920 TI - The role of qualitative research in evidence based practice. PMID- 12539921 TI - At the millennium: a slice of scholarship. AB - Scholarly output is the standard by which academics are judged; in particular journal articles. This study aimed to examine the scholarly output of nursing academics over a two-year period, and to establish benchmark data for future longitudinal comparisons. Journal articles were used as the unit of analysis. The following characteristics of articles were examined: whether articles were refereed or non-refereed; the type of journal; content characteristics; scholarly focus; predominant methodology of research articles; and predominant focus of specialty areas. The characteristics of principal authors and their effect on articles were also examined. The study demonstrated that while there had been no substantial increase in scholarly output, there was a clear link between higher academic rank and type of scholarly output. An increasing predominance of research articles was demonstrated, with a striking trend towards increased use of qualitative methodology, and in the number of publications by groups of authors. PMID- 12539923 TI - Exploring nursing: a conversation about nursing management. Interview by Jane Etchells. PMID- 12539922 TI - Nurse practitioners: a contract for change and excellence in nursing. AB - The role of the Nurse Practitioner has been in existence in a variety of contexts and within a broad range of the scope of their practice throughout the world for a number of years. Many nurses work at this advanced level of clinical practice without the acknowledgement f the very important and responsible role that they play within the healthcare setting. Although the United States and United Kingdom have recognised the role of the advanced Nurse Practitioner for a number of years, there still exists confusion and disagreement as to their scope of practice. There is uncertainty and anxiety as to where the role boundaries between nursing and medical and allied health professionals begin and end. The role of the Nurse Practitioner in Australia has not been without its problems in the developmental stage of its creation. New South Wales finally achieved recognition of the role this year after a decade of negotiation. This has culminated in the acceptance for the development of 40 Nurse Practitioner positions across the State. The first of these was accepted in the Far West Area Health Service in May 2001. The Far West Area Health Service created a five-year plan, which addresses the development of nurses preparing for authorisation, the creation of Nurse Practitioner positions in the remote communities, the creation of clinical guidelines to support advanced practice and the evaluation process for both the positions and the nurses. The objective of this approach is to ensure effective implementation of these advanced nursing positions in the remote communities of New South Wales. The Nurse Practitioner role needs to respond to the individual, the family and the community, utilising advanced clinical skills and remaining responsive to the changes in health care within a primary health care framework, which is essential for combating the complex health care issues in remote areas (NSW Health 2000). PMID- 12539924 TI - Nursing shortages: let's be flexible. AB - The following paper is a report outlining a significant work pattern change in an acute cardiac ward at a large Brisbane-based private hospital. The nursing staff expressed the desire for more flexible rostering and the opportunity to work 12 hour shifts. After agreement was reached between the hospital, the union and the Industrial Relations Board, guidelines were put in place and a Flexible Rostering System was proposed and trialed. An 80% consensus of staff was required both to proceed with the trial and to implement any permanent changes. Initially, the trial was conducted for three months and extended to six months. The shifts trialed were between four and 12 hours in length with varied starting and finishing times. The Flexible Rostering System was evaluated using feedback from staff surveys and the results of a staff vote. In addition, patient feedback, incident reports, financial and managerial evaluation of staff costs, hours per patient day utilised, sick leave, and the use of permanent staff for voluntary extra shifts were also monitored. The outcome of the trial was positive with over 80% of staff voting to implement the Flexible Rostering System permanently. A significant reduction in sick leave of 41% and improved retention of skilled registered nursing staff was noted. There was no increase in the number of incident reports or patient complaints. Both patients and nurses commented on the improved continuity of care. Salaries and wages were within budget. Staff surveys showed positive feedback such as increased morale, increased flexibility with rosters, decreased fatigue levels, improved patient assessment on night duty and an increase in number of days off. In conclusion, the Flexible Rostering System has been accepted as a positive change for staff and is cost effective for the hospital. In light of nursing shortages, the outcome of this trial cannot be ignored. PMID- 12539925 TI - Confusion prevails in defining 'advanced' nursing practice. AB - The Australian nursing profession has accepted the challenge to support and encourage advanced nursing practice role development. A critical review of national and international literature highlights that there is no singular definition or understanding of 'advanced' nursing practice. Instead difficulty with nomenclature and clarification of levels of nursing are perpetuated by unclear distinction between 'advanced', 'specialist' and 'expert' nursing practice. This paper presents the findings of a review of the literature and posits that ambiguity and lack of consistency in defining 'advanced' nursing practice is hampering the progression of the profession towards the development of consistent advanced practice roles in Australia. It is proposed that a universally accepted definition would provide a vehicle for future Australian role development so that all advanced practice roles have a common genesis and national consistency results. PMID- 12539926 TI - Male rape--the silent victims. AB - This literature review examines the issue of male rape and the possible counselling strategies that may assist in recovery. There are several misconceptions surrounding male rape which can result in the under-reporting and secondary or sanctuary victimisation of the survivor. Men who have been raped may believe that it attacks the very essence of what it is to be masculine and male. Many may not seek help unless they perceive a need for immediate attention, such as physical trauma requiring medical assistance. The literature also reveals that when men do seek help they may be treated poorly. Secondary victimisation or sanctuary trauma results when there is a lack of empathy and understanding of the effect that rape can have on the survivor, such as rape-trauma syndrome. Training in this area is needed for the police, emergency department staff, nurses, general practitioners, and community health services. Management of survivors starts with examining our own beliefs about male rape. Many of the reported counselling strategies are based on therapists' observations, trauma theories, and research related to child abuse or sexual assault of females which may not be transferable to men who have been raped. Education and counselling of survivors' support networks must be considered as part of a holistic approach to management. Secondary prevention strategies aimed at the broader community are required to assist survivors to come forward without fear of further victimisation because of stereotyping. PMID- 12539927 TI - Preoperative fasting procedures: let's use the evidence. PMID- 12539928 TI - Responses to olfactory stimuli in spotted hyenas (Crocuta crocuta): I. Investigation of environmental odors and the function of rolling. AB - Olfaction is crucial to spotted hyenas (Crocuta crocuta), yet there are no controlled studies of their reactions to odors. In Experiment 1, the authors examined responses of captive hyenas to various environmental (prey, nonprey animal, and plant) odors. Subjects approached and sniffed all odors equally but preferentially licked prey odors, scent marked next to odors, and rolled in animal-based odors. In Experiment 2, the authors examined the function of rolling by applying odors to the pelts of captive hyenas. When hyenas wore carrion, they gained positive social attention (increased investigation and allogrooming) from pen mates, but when they wore camphor, the normal social greeting ceremony was curtailed. Thus, olfactory stimuli elicit specific responses, influence where behavior is directed, and can be used to affect social interaction. PMID- 12539929 TI - Responses to olfactory stimuli in spotted hyenas (Crocuta crocuta): II. Discrimination of conspecific scent. AB - Scent marking in spotted hyenas (Crocuta crocuta) includes the deposition of anal sac secretions, or "paste," and presumably advertises territorial ownership. To test whether captive hyenas classify and discriminate individuals using odor cues in paste, the authors conducted behavioral discrimination bioassays and recorded hyena investigation of paste extracted from various conspecific donors. In Experiment 1, subjects directed most investigative behavior toward scents from unfamiliar hyenas and members of the opposite sex. In Experiment 2, male hyenas discriminated between concurrent presentations of paste from various unfamiliar females in similar reproductive states. Thus, pasted scent marks convey information about the sex, familiarity, and even identity of conspecifics. Aside from territory maintenance, scent marking may also communicate information about individual sexual status. PMID- 12539930 TI - A comparative study of geometric rule learning by nutcrackers (Nucifraga columbiana), pigeons (Columba livia), and jackdaws (Corvus monedula). AB - Three avian species, a seed-caching corvid (Clark's nutcrackers; Nucifraga columbiana), a non-seed-caching corvid (jackdaws; Corvus monedula), and a non seed-caching columbid (pigeons; Columba livia), were tested for ability to learn to find a goal halfway between 2 landmarks when distance between the landmarks varied during training. All 3 species learned, but jackdaws took much longer than either pigeons or nutcrackers. The nutcrackers searched more accurately than either pigeons or jackdaws. Both nutcrackers and pigeons showed good transfer to novel landmark arrays in which interlandmark distances were novel, but inconclusive results were obtained from jackdaws. Species differences in this spatial task appear quantitative rather than qualitative and are associated with differences in natural history rather than phylogeny. PMID- 12539931 TI - Short-term contact elicits heterospecific behavioral discrimination of individual odors in mound-building mice (Mus spicilegus). AB - The authors used a habituation-dishabituation procedure to test the ability of male mound-building mice (Mus spicilegus) to discriminate individual odors from males of another species of mouse. Male mound-building mice failed to spontaneously discriminate individual odors from Mus musculus musculus males, a natural competitor. After 24-hr contact with a male of one of the M. musculus subspecies (M. m. musculus or M. m. domesticus), experienced M. spicilegus males discriminated the individual odors of unfamiliar males of the same subspecies. These results confirm that discrimination of individual chemosignals is not confined to olfactory cues of a single species and provide new information about the effect of short-term contact on discrimination of individual odors across species. PMID- 12539932 TI - Socially induced delayed reproduction in female Mongolian gerbils (Meriones unguiculatus): is there anything special about dominant females? AB - In 4 experiments, the authors explored effects of interaction with both sexually active adult female and unfamiliar adult male Mongolian gerbils (Meriones unguiculatus) on young female gerbils' ages at first parturition. Presence of a natural mother retarded development of her daughters. However, presence of a natural mother had no greater effect on her daughters' development than did presence of any other familiar animal, either male or female. Further, exposing young female gerbils to an unfamiliar male accelerated their development even when their reproductively active mothers were present. The data indicate that maintaining young female gerbils in stable family groups results in both inhibition of sexual maturation (caused by exposure to familiar individuals) and failure to activate sexual development (caused by lack of exposure to unfamiliar males). PMID- 12539933 TI - Early visual experience affects postnatal auditory responsiveness in bobwhite quail (Colinus virginianus). AB - Results from 6 experiments suggest perinatal visual experience interferes with postnatal auditory responsiveness in bobwhite quail (Colinus virginianus). Light- or dark-reared control chicks responded similarly to bobwhite maternal Calls A or B following hatching. Light-reared chicks that experienced 10 min/hr of Call A or B from hatching until testing preferred the familiar call at Days 2-4, but dark reared chicks preferred the familiar call at all ages. Increased amounts of exposure to maternal calls during postnatal Days 1-2 led to auditory responsiveness deficits in light-reared chicks. Similarly, embryos exposed to 10 min/hr of prenatal visual cues required 48 hr of postnatal Call A or B exposure to demonstrate auditory discrimination. These findings highlight the linkages between developing auditory and visual systems during infancy. PMID- 12539934 TI - Temporal integration as a function of signal and gap intensity in rats (Rattus norvegicus) and pigeons (Columba livia). AB - Previous data suggest that rats (Rattus norvegicus) and pigeons (Columba livia) use different interval-timing strategies when a gap interrupts a to-be-timed signal: Rats stop timing during the gap, and pigeons reset their timing mechanism after the gap. To examine whether the response rule is controlled by an attentional mechanism dependent on the characteristics of the stimuli, the authors manipulated the intensity of the signal and gap when rats and pigeons timed in the gap procedure. Results suggest that both rats and pigeons stop timing during a nonsalient gap and reset timing after a salient gap. These results also suggest that both species use similar interval-timing mechanisms, influenced by nontemporal characteristics of the signal and gap. PMID- 12539935 TI - Multiple-landmark piloting in pigeons (Columba livia): landmark configuration as a discriminative cue. AB - Using the landmark-transformation technique, researchers have shown that pigeons (Columba livia) tend to encode a goal location relative to 1 landmark, even when multiple landmarks are in the vicinity of the goal. The current experiments examined pigeons' ability to use configural information from a set of landmarks by making the arrangement of 4 landmarks a discriminative cue to the location of buried seeds. Results showed that pigeons used information from the 3 consistently placed landmarks to search accurately when 1 landmark was displaced. Findings indicate that pigeons are able to search for a goal using information from multiple landmarks instead of just 1 and that landmark use by these birds may be more flexible than previously theorized. PMID- 12539936 TI - Lumbar spinal stenosis: surgical considerations. AB - Spinal stenosis is an acquired or congenital narrowing of the spinal or nerve root canals. Surgical treatment is often effective. Acquired spinal stenosis most commonly occurs in those with degenerative disk disease and arthritic facets. If the degenerative process stabilizes and there is adequate room to accommodate the neural contents, symptomatic patients become asymptomatic. Residual stability after decompression must be assessed in patients having multilevel decompression. Fusion maybe indicated. In women with osteoporosis coexisting with degenerative scoliosis and spinal stenosis, decompression for concave nerve-root compression and fusion are necessary. Spinal fusion is not indicated in patients with lumbar spinal stenosis having unilateral decompression for lateral stenosis. Patients with central-mixed stenosis may not need fusion. Patients with spinal stenosis after laminectomies and diskectomies had better results when arthrodesis was done in conjunction with repeated decompression. Arthrodesis with instrumentation and decompression is recommended for patients with degenerative spondylolisthesis. PMID- 12539937 TI - The Cooper River Bridge Run Study of low back pain in runners and walkers. AB - Investigations dedicated to examining relationships between low back pain (LBP) and factors specific to running and walking are limited. Current literature suggests runners and walkers may have a lower relative risk for significant LBP. The purpose of this study was to investigate the incidence, prevalence, and possible risk factors for LBP among a group of runners and walkers. A survey was administered to participants who registered for either a 10-kilometer run or 4 mile recreational walk. A total of 539 responses were received. Previous history of LBP was reported by 74% of respondents overall. Prevalence of LBP at the time of survey completion was 13.6%. Low back pain was experienced more frequently by obese runners and by those who reported certain patterns of shoe wear. Regular participation in aerobics correlated with a reduced lifetime risk for LBP. We believe these findings warrant further investigation. This knowledge may benefit not only athletes in training, but other fitness enthusiasts, including casual runners and walkers and those who jog or walk to rehabilitate low back injuries. PMID- 12539938 TI - Carpal tunnel release: efficacy and recurrence rate after a limited incision release. AB - We retrospectively studied the postoperative outcomes of 51 patients treated for idiopathic carpal tunnel syndrome by method of a limited incision carpal tunnel release. Patients were assessed to determine: 1) palmar tenderness, 2) scar tenderness, 3) relief of symptoms, 4) complications, and 5) recurrence. Short term follow-up included patient evaluations at 2 weeks, 4 weeks, and 10 weeks; a mean of 2.5 years of follow-up also was obtained. Postoperatively, nocturnal symptoms resolved by the 2-week visit. Palmar tenderness was noted as minimal or absent between the 4-week and 10-week visits in 47 of the 51 patients (92%). Symptom and function scores improved from 4.24 and 4.00 preoperatively to 1.18 and 1.19 postoperatively. At a mean of 2.5 years after surgery, none of the patients reported recurrent symptoms. PMID- 12539940 TI - Results of simultaneous bilateral total knee replacement: a study of 1208 knees in 604 patients. AB - Bilateral symptomatic knee arthritis is a common clinical problem. There are conflicting opinions as to the advisability of simultaneous sequential bilateral total knee replacement. Complication rates of primary unilateral knee replacement are well documented and there are several small series that compare the two techniques. The objective of this study was to identify the complication rate of simultaneous sequential bilateral total knee replacement in a large patient population. Over a 13-year period, 604 primary bilateral sequential simultaneous total knee replacements (1208 knees) were performed. Office notes and hospital charts were retrospectively reviewed to obtain age, sex, diagnosis, knee alignment, associated comorbidities, operative protocol, transfusions, and complications. The study results showed 5.1% local and 15.3% systemic complications and 0.7% mortality rate (none in the past 9 years). With appropriate patient selection and operative technique, patients who present with bilateral symptomatic knee arthritis can enjoy the benefits of simultaneous sequential bilateral total knee replacement without increasing their risks of complications. PMID- 12539939 TI - Biomechanical evaluation of fixation of clavicular fractures. AB - Reproducible fractures of the midshaft of the clavicle were created in 14 fresh frozen human cadaveric clavicles. Under the three-point bending loading by a materials testing machine, the load to failure of fixation of the clavicular fractures treated with steel reconstruction plates (five specimens) and Herbert cannulated bone screw (nine specimens), was compared with a group control formed by five specimens of clavicles without osteosynthesis material. No statistically significant differences were observed between the three groups. In consequence and in terms of biomechanics, in clavicular acute fractures, both plating and intramedullary Herbert cannulated bone screw may be utilized in the treatment of these lesions. PMID- 12539941 TI - Outcomes of posterolateral versus BAK titanium cage interbody lumbar fusion in injured workers: a retrospective cohort study. AB - Lumbar fusion has been criticized for variable patient outcomes, though little is known regarding how outcomes vary across procedures. We examined outcomes of posterolateral versus BAK interbody lumbar fusion in workers' compensation cases. A medical record review and a follow-up survey were completed. The sample included 185 posterolateral and 185 lumbar interbody fusions. An outcome survey was conducted an average of 5 years after surgery. Arthrodesis rates, satisfaction, function, and health were better for the BAK interbody lumbar fusion cohort. Results suggest greater efficacy of the BAK interbody approach over posterolateral approaches to lumbar fusion in compensated patients. PMID- 12539942 TI - Biostructural augmentation for the treatment of osteonecrosis: rationale, technique, and case example. AB - Avascular necrosis of the hip is a common disease that usually affects a young and active patient population. As the disease progresses, the undermined structural integrity of the subchondral bone leads to articular collapse and subsequent osteoarthrosis. The ideal treatment is one that hinders or arrests the progression of the disease, averting articular collapse and joint replacement surgery. A surgical strategy is described that attempts to address the multiple factors involved in the progression of the disease. This is accomplished through a modified core decompression procedure combined with the insertion of two interference screws into the subchondral plate to provide structural support and the use of osteoinductive material (i.e., demineralized bone matrix) in an effort to accelerate the bone healing process. PMID- 12539943 TI - Multifocal osteonecrosis and human immunodeficiency virus infection. AB - Over the past decade, a causal relationship between infection with human immunodeficiency virus (HIV) and osteonecrosis has been proposed. This case report (the first example presented in the English orthopaedic literature) of multifocal (> 3 sites) osteonecrosis in a patient with HIV infection highlights the current scientific knowledge regarding incidence, predisposing risk factors, proposed biologic mechanisms, and clinical orthopaedic management. This information may become increasingly relevant for the orthopaedic clinician as more patients with HIV infection live longer. PMID- 12539945 TI - 'In silico' simulation of biological processes. Chair's introduction. PMID- 12539944 TI - The pink pulseless hand. AB - The management of a child with a perfused, pink, but pulseless upper extremity following reduction and pinning of a type III supracondylar humerus fracture remains controversial. The authors present the initial treatment, evaluation, operative findings, and postoperative course of a 6-year-old with a pink pulseless hand. Review of the literature is included, as well as recommendations regarding operative management. PMID- 12539946 TI - Integrative biological modelling in silico. AB - In silico models of biological systems provide a powerful tool for integrative analysis of physiological function. Using the computational models of the heart as examples, we discuss three types of integration: structural integration implies integration across physical scales of biological organization from protein molecule to whole organ; functional integration of interacting physiological processes such as signalling, metabolism, excitation and contraction; and the synthesis of experimental observation with physicochemical and mathematical principles. PMID- 12539947 TI - Advances in computing, and their impact on scientific computing. AB - This paper begins by discussing the developments and trends in computer hardware, starting with the basic components (microprocessors, memory, disks, system interconnect, networking and visualization) before looking at complete systems (death of vector supercomputing, slow demise of large shared-memory systems, rapid growth in very large clusters of PCs). It then considers the software side, the relative maturity of shared-memory (OpenMP) and distributed-memory (MPI) programming environments, and new developments in 'grid computing'. Finally, it touches on the increasing importance of software packages in scientific computing, and the increased importance and difficulty of introducing good software engineering practices into very large academic software development projects. PMID- 12539948 TI - From physics to phenomenology. Levels of description and levels of selection. AB - Formal models in biology are traditionally of two types: simulation models in which individual components are described in detail with extensive empirical support for parameters, and phenomenological models, in which collective behaviour is described in the hope of identifying critical variables and parameters. The advantage of simulation is greater realism but at a cost of limited tractability, whereas the advantage of phenomenological models, is greater tractability and insight but at a cost of reduced predictive power. Simulation models and phenomenological models lie on a continuum, with phenomenological models being a limiting case of simulation models. I survey these two levels of model description in genetics, molecular biology, immunology and ecology. I suggest that evolutionary considerations of the levels of selection provides an important justification for many phenomenological models. In effect, evolution reduces the dimension of biological systems by promoting common paths towards increased fitness. PMID- 12539949 TI - Making sense of complex phenomena in biology. AB - The remarkable advances in biotechnology over the past two decades have resulted in the generation of a huge amount of experimental data. It is now recognized that, in many cases, to extract information from this data requires the development of computational models. Models can help gain insight on various mechanisms and can be used to process outcomes of complex biological interactions. To do the latter, models must become increasingly complex and, in many cases, they also become mathematically intractable. With the vast increase in computing power these models can now be numerically solved and can be made more and more sophisticated. A number of models can now successfully reproduce detailed observed biological phenomena and make important testable predictions. This naturally raises the question of what we mean by understanding a phenomenon by modelling it computationally. This paper briefly considers some selected examples of how simple mathematical models have provided deep insights into complicated chemical and biological phenomena and addresses the issue of what role, if any, mathematics has to play in computational biology. PMID- 12539950 TI - On ontologies for biologists: the Gene Ontology--untangling the web. AB - The mantra of the 'post-genomic' era is 'gene function'. Yet surprisingly little attention has been given to how functional and other information concerning genes is to be captured, made accessible to biologists or structured in a computable form. The aim of the Gene Ontology (GO) Consortium is to provide a framework for both the description and the organisation of such information. The GO Consortium is presently concerned with three structured controlled vocabularies which can be used to describe three discrete biological domains, building structured vocabularies which can be used to describe the molecular function, biological roles and cellular locations of gene products. PMID- 12539951 TI - The KEGG database. AB - KEGG (http://www.genome.ad.jp/kegg/) is a suite of databases and associated software for understanding and simulating higher-order functional behaviours of the cell or the organism from its genome information. First, KEGG computerizes data and knowledge on protein interaction networks (PATHWAY database) and chemical reactions (LIGAND database) that are responsible for various cellular processes. Second, KEGG attempts to reconstruct protein interaction networks for all organisms whose genomes are completely sequenced (GENES and SSDB databases). Third, KEGG can be utilized as reference knowledge for functional genomics (EXPRESSION database) and proteomics (BRITE database) experiments. I will review the current status of KEGG and report on new developments in graph representation and graph computations. PMID- 12539952 TI - Bioinformatics of cellular signalling. AB - The completion of the human genome sequencing provides a unique opportunity to understand the complex functioning of cells in terms of myriad biochemical pathways. Of special significance are pathways involved in cellular signalling. Understanding how signal transduction occurs in cells is of paramount importance to medicine and pharmacology. The major steps involved in deciphering signalling pathways are: (a) identifying the molecules involved in signalling; (b) figuring out who talks to whom, i.e. deciphering molecular interactions in a context specific manner; (c) obtaining the spatiotemporal location of the signalling events; (d) reconstructing signalling modules and networks evoked in specific response to input; (e) correlating the signalling response to different cellular inputs; and (f) deciphering cross-talk between signalling modules in response to single and multiple inputs. High-throughput experimental investigations offer the promise of providing data pertaining to the above steps. A major challenge, then, is the organization of this data into knowledge in the form of hypothesis, models and context-specific understanding. The Alliance for Cellular Signaling (AfCS) is a multi-institution, multidisciplinary project and its primary objective is to utilize a multitude of high throughput approaches to obtain context-specific knowledge of cellular response to input. It is anticipated that the AfCS experimental data in combination with curated gene and protein annotations, available from public repositories, will serve as a basis for reconstruction of signalling networks. It will then be possible to model the networks mathematically to obtain quantitative measures of cellular response. In this paper we describe some of the bioinformatics strategies employed in the AfCS. PMID- 12539953 TI - Imaging-based integrative models of the heart: closing the loop between experiment and simulation. AB - We describe methodologies for: (a) mapping ventricular activation using high density epicardial electrode arrays; (b) measuring and modelling ventricular geometry and fibre orientation at high spatial resolution using diffusion tensor magnetic resonance imaging (DTMRI); and (c) simulating electrical conduction; using comprehensive data sets collected from individual canine hearts. We demonstrate that computational models based on these experimental data sets yield reasonably accurate reproduction of measured epicardial activation patterns. We believe this ability to electrically map and model individual hearts will lead to enhanced understanding of the relationship between anatomical structure, and electrical conduction in the cardiac ventricles. PMID- 12539954 TI - The Virtual Cell project. AB - The Virtual Cell is a modular computational framework that permits construction of models, application of numerical solvers to perform simulations, and analysis of simulation results. A key feature of the Virtual Cell is that it permits the incorporation of realistic experimental geometries within full 3D spatial models. An intuitive JAVA interface allows access via a web browser and includes options for database access, geometry definition (including directly from microscope images), specification of compartment topology, species definition and assignment, chemical reaction input and computational mesh. The system is designed for cell biologists to aid both the interpretation and the planning of experiments. It also contains sophisticated modelling tools that are appropriate for the needs of mathematical biologists. Thus, communication between these traditionally separate scientific communities can be facilitated. This paper will describe the status of the project and will survey several applications to cell biological problems. PMID- 12539955 TI - Modelling the bacterial chemotaxis receptor complex. AB - The pathway controlling chemotaxis in Escherichia coli is the simplest and most well understood cell signalling system to date. However, quantitative models based on the available data still fail to reproduce important features of the pathway. Most notably, the observed sensitivity of cells to very small changes in stimulus concentrations cannot be reproduced by conventional models based on the measured concentrations, binding affinities and rate constants of the proteins involved. This discrepancy, together with recent experimental findings, drew our attention to the spatial organization of molecules within the cell and in particular to the clusters of receptors localised at the cell poles. A stochastic simulator for chemical reactions, STOCHSIM, was previously developed to model the chemotaxis pathway at the level of individual molecular interactions. This program has now been extended to incorporate a spatial representation that allows the interaction between molecules in a two-dimensional lattice to be simulated. In silico 'experiments' using this new version of STOCHSIM demonstrate that lateral interactions between clustered receptors can significantly enhance the excitation response. The adaptation reactions may also exploit the proximity of receptor molecules, and a hypothetical mechanism by which this may occur is currently being tested. PMID- 12539956 TI - The heart cell in silico: successes, failures and prospects. AB - The development of computer models of heart cells is used to illustrate the interaction between simulation and experimental work. At each stage, the reasons for new models are explained, as are their defects and how these were used to point the way to successor models. As much, if not more, was learnt from the way in which models failed as from their successes. The insights gained are evident in the most recent developments in this field, both experimental and theoretical. The prospects for the future are discussed. PMID- 12539957 TI - The IUPS Physiome Project. International Union of Physiological Sciences. AB - Modern medicine is currently benefiting from the development of new genomic and proteomic techniques, and also from the development of ever more sophisticated clinical imaging devices. This will mean that the clinical assessment of a patient's medical condition could, in the near future, include information from both diagnostic imaging and DNA profile or protein expression data. The Physiome Project of the International Union of Physiological Sciences (IUPS) is attempting to provide a comprehensive framework for modelling the human body using computational methods which can incorporate the biochemistry, biophysics and anatomy of cells, tissues and organs. A major goal of the project is to use computational modelling to analyse integrative biological function in terms of underlying structure and molecular mechanisms. To support that goal the project is establishing web-accessible physiological databases dealing with model-related data, including bibliographic information, at the cell, tissue, organ and organ system levels. This paper discusses the development of comprehensive integrative mathematical models of human physiology based on patient-specific quantitative descriptions of anatomical structures and models of biophysical processes which reach down to the genetic level. PMID- 12539958 TI - Using in silico biology to facilitate drug development. AB - G protein-coupled receptor (GPCR) mediation of cardiac excitability is often overlooked in predicting the likelihood that a compound will alter repolarization. While the areas of GPCR signal transduction and electrophysiology are rich in data, experiments combining the two are difficult. In silico modelling facilitates the integration of all relevant data in both areas to explore the hypothesis that critical associations may exist between the different GPCR signalling mechanisms and cardiac excitability and repolarization. An example of this linkage is suggested by the observation that a mutation of the gene encoding HERG, the pore-forming subunit of the rapidly activating delayed rectifier K+ current (I(Kr)), leads to a form of long QT syndrome in which affected individuals are vulnerable to stress-induced arrhythmia following beta adrenergic stimulation. Using Physiome's In Silico Cell, we constructed a model integrating the signalling mechanisms of second messengers cAMP and protein kinase A with I(Kr) in a cardiac myocyte. We analysed the model to identify the second messengers that most strongly influence I(Kr) behaviour. Our conclusions indicate that the dynamics of regulation are multifactorial, and that Physiome's approach to in silico modelling helps elucidate the subtle control mechanisms at play. PMID- 12539959 TI - Transport ATPases in biological systems and relationship to human disease: a brief overview. AB - Interest in the field of transport ATPases has grown dramatically during the past 20 years and gained considerable visibility for several reasons. First, it was shown that most transport ATPases can be lumped into only a few categories designated simply as P, V, F, and ABC types, the latter consisting of a large superfamily. Second, it has been shown that many transport ATPases have a clear relevance to human disease. Third, the field of transport ATPases has become rather advanced in the study of the reaction mechanisms and structure-function relationships associated with several of these enzymes. Finally, the Nobel committee recently recognized major accomplishments in this field of research. Here, the author provides a brief discussion of transport ATPases that are present in biological systems and their relevance or possible relevance to human disease. PMID- 12539960 TI - Copper transporting P-type ATPases and human disease. AB - Copper transporting P-type ATPases, designated ATP7A and ATP7B, play an essential role in mammalian copper balance. Impaired intestinal transport of copper, resulting from mutations in the ATP7A gene, lead to Menkes disease in humans. Defects in a similar gene, the copper transporting ATPase ATP7B, result in Wilson disease. This ATP7B transporter has two functions: transport of copper into the plasma protein ceruloplasmin, and elimination of copper through the bile. Variants of ATP7B can be functionally assayed to identify defects in each of these functions. Tissue expression studies of the copper ATPases and their copper chaperone ATOX1 indicate that there is not complete overlap in expression. Other chaperones may be important for the transport of copper into ATP7A and ATP7B. PMID- 12539961 TI - Structural and functional insights of Wilson disease copper-transporting ATPase. AB - Wilson disease is an autosomal recessive disorder of copper metabolism. The gene for this disorder has been cloned and identified to encode a copper-transporting ATPase (ATP7B), a member of a large family of cation transporters, the P-type ATPases. In addition to the core elements common to all P-type ATPases, the Wilson copper-transporting ATPase has a large cytoplasmic N-terminus comprised six heavy metal associated (HMA) domains, each of which contains the copper binding sequence motif GMT/HCXXC. Extensive studies addressing the functional, regulatory, and structural aspects of heavy metal transport by heavy metal transporters in general, have offered great insights into copper transport by Wilson copper-transporting ATPase. The findings from these studies have been used together with homology modeling of the Wilson disease copper-transporting ATPases based on the X-ray structure of the sarcoplasmic reticulum (SR) calcium-ATPase, to present a hypothetical model of the mechanism of copper transport by copper transporting ATPases. PMID- 12539962 TI - Human copper-transporting ATPase ATP7B (the Wilson's disease protein): biochemical properties and regulation. AB - Wilson's disease protein (WNDP) is a product of a gene ATP7B that is mutated in patients with Wilson's disease, a severe genetic disorder with hepatic and neurological manifestations caused by accumulation of copper in the liver and brain. In a cell, WNDP transports copper across various cell membranes using energy of ATP-hydrolysis. Copper regulates WNDP at several levels, modulating its catalytic activity, posttranslational modification, and intracellular localization. This review summarizes recent studies on enzymatic function and copper-dependent regulation of WNDP. Specifically, we describe the molecular architecture and major biochemical properties of WNDP, discuss advantages of the recently developed functional expression of WNDP in insect cells, and summarize the results of the ligand-binding studies and molecular modeling experiments for the ATP-binding domain of WNDP. In addition, we speculate on how copper binding may regulate the activity and intracellular distribution of WNDP, and what role the human copper chaperone Atox1 may play in these processes. PMID- 12539963 TI - Menkes copper-translocating P-type ATPase (ATP7A): biochemical and cell biology properties, and role in Menkes disease. AB - The Menkes copper-translocating P-type ATPase (ATP7A; MNK) is a ubiquitous protein that regulates the absorption of copper in the gastrointestinal tract. Inside cells the protein has a dual function: it delivers copper to cuproenzymes in the Golgi compartment and effluxes excess copper. The latter property is achieved through copper-dependent vesicular trafficking of the Menkes protein to the plasma membrane of the cell. The trafficking mechanism and catalytic activity combine to facilitate absorption and intercellular transport of copper. The mechanism of catalysis and copper-dependent trafficking of the Menkes protein are the subjects of this review. Menkes disease, a systemic copper deficiency disorder, is caused by mutations in the gene encoding the Menkes protein. The effect of these mutations on the catalytic cycle and the cell biology of the Menkes protein, as well as predictions of the effect of particular mutant MNKs on observed Menkes disease symptoms will also be discussed. PMID- 12539964 TI - Copper chaperones: personal escorts for metal ions. AB - Copper serves as the essential cofactor for a number of enzymes involved in redox chemistry and virtually all organisms must accumulate trace levels of copper in order to survive. However, this metal can also be toxic and a number of effective methods for sequestering and detoxifying copper prevent the metal from freely circulating inside a cell. Copper metalloenzymes are therefore faced with the challenge of acquiring their precious metal cofactor in the absence of available copper. To overcome this dilemma, all eukaryotic organisms have evolved with a family of intracellular copper binding proteins that help reserve a bioavailable pool of copper for the metalloenzymes, escort the metal to appropriate targets, and directly transfer the copper ion. These proteins have been collectively called "copper chaperones." The identification of such molecules has been made possible through molecular genetic studies in the bakers' yeast Saccharomyces cerevisiae. In this review, we highlight the findings that led to a new paradigm of intracellular trafficking of copper involving the action of copper chaperones. In particular, emphasis will be placed on the ATX1 and CCS copper chaperones that act to deliver copper to the secretory pathway and to Cu/Zn superoxide dismutase in the cytosol, respectively. PMID- 12539966 TI - Role of copper in mitochondrial biogenesis via interaction with ATP synthase and cytochrome c oxidase. AB - Animals that are copper deficient have cardiac hypertrophy where there is a dramatic increase in mitochondria. Mitochondrial biogenesis is enhanced in this model and there is an upregulation of mitochondrial transcription factor A (mtTFA) and nuclear respiratory factors 1 and 2 (NRF-1 and NRF-2). While the cuproenzyme, cytochrome c oxidase (CCO), is an attractive candidate to explain the connection between cardiac hypertrophy in copper deficiency and subsequent mitochondrial biogenesis, studies have revealed that ATP synthase may be impacted by copper depletion. NRF-1 and NRF-2 can bind to some of the subunits of both CCO and ATP synthase to regulate gene expression. Furthermore, oxidative phosphorylation appears to occur unaltered in the copper-deficient state. Copper deficient mitochondria appear to be less sensitive to the inhibitory effect of oligomycin compared to controls. Decreases in the delta subunit protein and beta mRNA transcript have been reported for ATP synthase as a function of copper deficiency. The limited data available suggest that copper, either indirectly or directly, alters ATP synthase function. PMID- 12539965 TI - Copper chaperones for cytochrome c oxidase and human disease. AB - Biological processes in living cells are compartmentalized between lipid membranes. Integral membrane proteins often confer specific functions to these compartments and as such have a critical role in cellular metabolism and function. Cytochrome c oxidase is a macromolecular metalloprotein complex essential for the respiratory function of the cell. Elucidating the mechanisms of assembly of cytochrome c oxidase within the inner mitochondrial membrane represents a unique challenge for understanding metalloprotein biosynthesis. Elegant genetic experiments in yeast have defined several proteins required for copper delivery to cytochrome c oxidase. While the precise role of each of these proteins in copper incorporation remains unclear, recent studies have revealed that inherited mutations in two of these proteins can result in severe pathology in human infants in association with cytochrome c oxidase deficiency. Characterization of the molecular pathogenesis of these disorders offers new insights into the mechanisms of cellular copper metabolism and the role of these cytochrome c oxidase copper chaperones in human disease. PMID- 12539967 TI - Impaired cardiac mitochondrial membrane potential and respiration in copper deficient rats. AB - Cardiac mitochondrial respiration, ATP synthase activity, and membrane potential and intactness were evaluated in copper-deficient rats. In the presence of NADH, both copper-deficient and copper-adequate mitochondria had very low oxygen consumption rates, indicating membrane intactness. However copper-deficient mitochondria had significantly lower oxygen consumption rates with NADH than did copper-adequate mitochondria. Copper-deficient mitochondria had significantly lower membrane potential than did copper-adequate mitochondria using fluorescent dyes. Copper-deficient mitochondria had significantly lower state 3 oxygen consumption rates and were less sensitive to inhibition by oligomycin, an ATP synthase inhibitor. Copper-deficient and copper-adequate mitochondria responded similiarly to CCCP. No difference was observed in mitochondrial ATPase activity between copper-deficient and copper-adequate rats using submitochondrial particles. We conclude that cardiac mitochondrial respiration is compromised in copper-deficient rats, and may be related to an altered ATP synthase complex and/or a decreased mitochondrial membrane potential. PMID- 12539970 TI - Expanding the frequency range of the solid-state T1rho experiment for heteronuclear dipolar relaxation. AB - Solid-state spin-lattice relaxation in the rotating frame permits the investigation of dynamic processes with correlation times in the range of microseconds. The relaxation process in organic solids is driven by the fluctuation of the local magnetic field due to the dipole-dipole interaction of the probe nuclei (13C,15N) with 1H in close proximity. However, its effect is often hidden by a competing relaxation process due to the contact between the rotating frame 13C/15N Zeeman and 1H dipolar reservoirs. In most cases the latter process becomes superior for the commonly applied low and moderate spin-lock fields and practically does not provide information about the molecular dynamics. To suppress this undesired process and to expand the dynamic range of T1rho experiments, we present two approaches. The first one uses a resonance offset of the frequency of the spin-lock irradiation, which leads to a significant enhancement of the effective spin-lock frequency without the application of destructive high transmitter powers. We derive the theory and demonstrate the applicability of the method on various model compounds. The second approach utilizes heteronuclear 1H decoupling during the 13C/15N spin-lock irradiation which disrupts the contact between the 13C/15N Zeeman and 1H dipolar reservoirs. We demonstrate the method and discuss the results qualitatively. PMID- 12539968 TI - Acalculia and dyscalculia. AB - Even though it is generally recognized that calculation ability represents a most important type of cognition, there is a significant paucity in the study of acalculia. In this paper the historical evolution of calculation abilities in humankind and the appearance of numerical concepts in child development are reviewed. Developmental calculation disturbances (developmental dyscalculia) are analyzed. It is proposed that calculation ability represents a multifactor skill, including verbal, spatial, memory, body knowledge, and executive function abilities. A general distinction between primary and secondary acalculias is presented, and different types of acquired calculation disturbances are analyzed. The association between acalculia and aphasia, apraxia and dementia is further considered, and special mention to the so-called Gerstmann syndrome is made. A model for the neuropsychological assessment of numerical abilities is proposed, and some general guidelines for the rehabilitation of calculation disturbances are presented. PMID- 12539969 TI - Hemisphere specialization as an aid in early infancy. AB - In order that different directions of attention can be organized, they have to be labeled and assessed. A statement of a general problem can be regarded as a label for a general direction of attention. Hope about it, as the perceived probability of sufficient success, on the basis of work done, can be regarded as an assessment. It can be argued that a young infant meets an impasse arising from the work on 2 incompatible general problems, (1) that of raising hope of certainty about the environment (linked to the arousal system because repeated stimulation has less effect), and (2) that of raising hope of producing effects (linked to the activation system because here some effect must be produced before activity can cease). A certainty-right hypothesis, that the right hemisphere deals with the certainty problem and the left deals with the producing-effects problem, and hence keeps work on the two problems apart in early infancy while the corpus callosum is undeveloped, and that a matching specialization continues in later life, is supported. PMID- 12539971 TI - The 13C chemical shift tensor principal values and orientations in dialkyl carbonates and trithiocarbonates. AB - The 13C chemical shift tensor principal values for the trigonal carbonate and thiocarbonate carbon atoms in the dialkyl carbonates, dimethyl carbonate, ethylene carbonate, and diphenyl carbonate, and in the trithiocarbonates, ethylene trithiocarbonate and dimethyl trithiocarbonate, respectively, were measured in various solid-state one-dimensional and two-dimensional nuclear magnetic resonance experiments. Furthermore, the chemical shift tensor principal values and orientations were calculated for the corresponding isolated molecules with quantum mechanically fully optimized geometries. Proton-optimized X-ray geometries of ethylene carbonate, ethylene trithiocarbonate, and diphenyl carbonate were used in embedded ion method (EIM) calculations and in calculations on the isolated molecules to obtain the theoretical principal values and to assign the chemical shift tensor orientations in these three compounds. Considerable improvement in the correlation between the experimental and calculated principal values is obtained when the electrostatic crystal potentials are included in EIM calculations. The chemical shift tensor orientations and principal values obtained for the dialkyl compounds in this study complement the previous data on a series of ionic potassium carbonates and thiocarbonates. PMID- 12539972 TI - Stimulated-echo NMR spectroscopy of 9Be and 7Li in solids: method and application to ion conductors. AB - The generation of pure quadrupolar stimulated-echo spectra is successfully demonstrated for the spin-3/2 probe 9Be in a single crystal of triglycine fluoberyllate. This solid exhibits a paraelectric-to-ferroelectric phase transition. From experiments carried out for various mixing times no indications for a slow soft mode could be detected in this crystal. Then ion conducting lithium metal phosphates were studied using 7Li, another spin-3/2 probe which allows for a non-selective excitation of the entire NMR spectrum. In the indium and the scandium phosphates ultra-slow Li hopping processes could be detected directly via the stimulated-echo technique in a time range of up to four orders of magnitude. Due to the relatively large gyromagnetic ratio and thus strong dipolar interactions of 7Li no pure quadrupolar echoes could be generated. However, from a variation of the evolution times the quadrupolar effects could be separated from the dipolar ones. Finally, the differences in the ion hopping times of lithium indium phosphate and of lithium scandium phosphate are briefly discussed. PMID- 12539973 TI - Double-resonance decoupling for resolution enhancement of 31P solid-state MAS and 27Al --> 31P MQHETCOR NMR. AB - 27Al decoupling has been used to remove residual J-coupling interactions between 31P and 27Al in microporous aluminophosphates AlPO4-14 and AlPO4-40. In combination with 1H high-power decoupling, 27Al adapted decoupling yields 31P spectra with optimal sensitivity and resolution. The importance of double resonance decoupling is further demonstrated by incorporating this technique in the MQHETCOR sequence. Unambiguous assignment of all the AlPO4-14 nuclear magnetic resonances is achieved by combining multiple-quantum evolution in the 27Al dimension and double-resonance decoupling in the 31P acquisition domain. PMID- 12539974 TI - Simultaneous molecular and anatomical imaging of the mouse in vivo. AB - Non-invasive imaging technologies are opening up new windows into mouse biology. We have developed a mouse imaging system that integrates positron emission tomography (PET) with x-ray computed tomography (CT), allowing simultaneous anatomic and molecular imaging in vivo with the potential for precise registration of the two image volumes. The x-ray system consists of a compact mini-focal x-ray tube and an amorphous selenium flat panel x-ray detector with a low-noise CMOS readout. The PET system uses planar arrays of lutetium oxyorthosilicate scintillator coupled to position-sensitive photomultiplier tubes. We describe the design of this dual-modality imaging system and show, for the first time, simultaneously acquired PET and CT images in a phantom and in mice. PMID- 12539975 TI - Statistical and heuristic image noise extraction (SHINE): a new method for processing Poisson noise in scintigraphic images. AB - Poisson noise is one of the factors degrading scintigraphic images, especially at low count level, due to the statistical nature of photon detection. We have developed an original procedure, named statistical and heuristic image noise extraction (SHINE), to reduce the Poisson noise contained in the scintigraphic images, preserving the resolution, the contrast and the texture. The SHINE procedure consists in dividing the image into 4 x 4 blocks and performing a correspondence analysis on these blocks. Each block is then reconstructed using its own significant factors which are selected using an original statistical variance test. The SHINE procedure has been validated using a line numerical phantom and a hot spots and cold spots real phantom. The reference images are the noise-free simulated images for the numerical phantom and an extremely high counts image for the real phantom. The SHINE procedure has then been applied to the Jaszczak phantom and clinical data including planar bone scintigraphy, planar Sestamibi scintigraphy and Tl-201 myocardial SPECT. The SHINE procedure reduces the mean normalized error between the noisy images and the corresponding reference images. This reduction is constant and does not change with the count level. The SNR in a SHINE processed image is close to that of the corresponding raw image with twice the number of counts. The visual results with the Jaszczak phantom SPECT have shown that SHINE preserves the contrast and the resolution of the slices well. Clinical examples have shown no visual difference between the SHINE images and the corresponding raw images obtained with twice the acquisition duration. SHINE is an entirely automatic procedure which enables halving the acquisition time or the injected dose in scintigraphic acquisitions. It can be applied to all scintigraphic images, including PET data, and to all low-count photon images. PMID- 12539976 TI - High resolution three-dimensional visualization and characterization of coronary atherosclerosis in vitro by synchrotron radiation x-ray microtomography and highly localized x-ray diffraction. AB - Human atherosclerotic plaques in both native and bypass arteries have been visualized using microtomography to provide additional information on the nature of coronary artery disease. Plaques contained within arteries removed from three white males aged 51, 55 and 70 are imaged in three-dimensions with monochromatic synchrotron x-ray radiation. Fields of view are 658 x 658 x 517 voxels. with cubic voxels ranging from 12 to 13 microm on a side. X-ray energies range from 11 to 15 keV (bandpass approximately 10 eV). At lower energies, high local absorption tends to generate reconstruction artefacts, while at higher energies the arterial wall is scarcely visible. At all energies, calcifications are clearly visible and differences are observed between plaques in native arteries (lifetime accumulations) versus bypass arteries (plaques developing in the interval between the heart bypass operation and the autopsy). In order to characterize coronary calcification, a microfocused, 50 microm2, 25 keV x-ray beam was used to acquire powder diffraction data from selected calcifications. Also, large calcifications were removed from the native arteries and imaged with 25 keV x-ray energy. Calcifications are composed of hydroxyapatite crystallites and an amorphous phase. In summary, native calcifications are larger and have a higher fraction of hydroxyapatite than calcifications from the bypass arteries. PMID- 12539977 TI - A Monte Carlo investigation of dual-planar circular-orbit cone-beam SPECT. AB - We use Monte Carlo simulations to study the imaging properties of a design for a dual-planar cone-beam (DPCB) single-photon emission computed tomography (SPECT) system. A dual-planar system uses a dual-camera SPECT system and two cone-beam collimators with foci in different axial planes to increase the effective axial field of view (FOV). We simulated nearly noise-free projection data from a computerized brain phantom and a phantom consisting of a series of points. Four configurations were simulated: parallel-beam low-energy high-resolution (LEHR) as a standard for comparison and DPCB at three radii of rotation (ROR) corresponding to the smallest, average and largest ROR required to clear patients' shoulders based on ergonomic data. We compared global measures of average resolution and total acquired counts for the four configurations. We also estimated local spatial frequency response for reconstructions of point sources. Finally, we estimated a local noise power spectrum by simulating 1000 noise realizations of the brain phantom and estimating a local noise covariance at selected points. The noise power spectra were used to estimate spectral signal to noise ratio (SNR) for each configuration. The resolution in the reconstructed image space ranges from 7.2 mm full-width at half-maximum (FWHM) at the minimum ROR to 9.4 mm FWHM at the maximum ROR. The efficiency is inversely related, ranging from 1.5 times that of parallel LEHR at minimum ROR to 2.5 times that of LEHR at maximum ROR. Estimates of system frequency response roughly correspond to the global resolution estimates, but the cone-beam techniques exhibit an unusual secondary peak in the axial-direction response. Estimates of spectral SNR show that the cone-beam configurations almost always result in higher SNR at all spatial frequencies regardless of ROR. The very largest ROR may be an exception. A larger ROR results in significantly higher SNR for low spatial frequencies with small reductions in SNR for mid-range frequencies. We conclude that the DPCB design allows significant improvements in both resolution and noise as compared to conventional parallel designs and that optimizing the ROR for the cone-beam system may improve the performance of certain imaging tasks. PMID- 12539978 TI - Portal dose image verification: formalism and application of the collapsed cone superposition method. AB - A formalism tailored for portal dose image verification is proposed to facilitate the comparison of calculated and measured portal dose distributions. Each portal image is converted into a dose proportional image and normalized to the reference beam calibration dose per monitor unit. The calculated or measured dose to a detector phantom is accordingly normalized so as to enable direct comparison. The collapsed cone kernel superposition method is adapted and evaluated for calculation of portal dose distributions in a water-equivalent detector phantom through comparisons with Monte Carlo calculations and with measurements. The deviation compared with Monte Carlo calculations for 6 and 15 MV was between +0.9% (the 0.9 quantile) and -2.1% (the 0.1 quantile) for a range of investigated geometries. Collapsed cone calculations compared with measurements for clinical fields agreed within [-1.9%, +2.4%] for 15 MV and [-0.9%, +3.2%] for 6 MV for the 0.1 and 0.9 quantiles, respectively. Hence, the absolute portal dose to a detector phantom could be calculated and verified well within the present accuracy requirements for clinical dose calculations. PMID- 12539979 TI - Manual multi-leaf collimator for electron beam shaping--a feasibility study. AB - In electron beam therapy, lead or low melting point alloy (LMA) sheet cutouts of sufficient thickness are commonly used to shape the beam. In order to avoid making cutouts for each patient, an attempt has been made to develop a manual multi-leaf collimator for electron beams (eMLC). The eMLC has been developed using LMA for a 15 x 15 cm2 applicator. Electron beam characteristics such as depth dose, beam profiles, surface dose, output factors and virtual source position with the eMLC have been studied and compared with those of an applicator electron beam. The interleaf leakage radiation has also been measured with film dosimetry. Depth dose values obtained using the eMLC were found to be identical to those with the applicator for depths larger than Dmax. However, a decrease in the size of the beam penumbra with the eMLC and increase in the values of surface dose, output factors and virtual source position with eMLC were observed. The leakage between the leaves was less than 5% and the leakage between the opposing leaves was 15%, which could be minimized further by careful positioning of the leaves. It is observed that it is feasible to use such a manual eMLC for patients and eliminate the fabrication of cutouts for each patient. PMID- 12539980 TI - Investigation of ultrasonic properties of PAG and MAGIC polymer gel dosimeters. AB - Ultrasonic speed of propagation and attenuation were investigated as a function of absorbed radiation dose in PAG and MAGIC polymer gel dosimeters. Both PAG and MAGIC gel dosimeters displayed a dependence of ultrasonic parameters on absorbed dose with attenuation displaying significant changes in the dose range investigated. The ultrasonic attenuation dose sensitivity at 4 MHz in MAGIC gels was determined to be 4.7 +/- 0.3 dB m(-1) Gy(-1) and for PAG 3.9 +/- 0.3 dB m(-1) Gy(-1). Ultrasonic speed dose sensitivities were 0.178 +/- 0.006 m s(-1) Gy(-1) for MAGIC gel and -0.44 +/- 0.02 m s(-1) Gy(-1) for PAG. Density and compressional elastic modulus were investigated to explain the different sensitivities of ultrasonic speed to radiation for PAG and MAGIC gels. The different sensitivities were found to be due to differences in the compressional elastic modulus as a function of dose for the two formulations. To understand the physical phenomena underlying the increase in ultrasonic attenuation with dose, the viscoelastic properties of the gels were studied. Results suggest that at ultrasonic frequencies, attenuation in polymer gel dosimeters is primarily due to volume viscosity. It is concluded that ultrasonic attenuation significantly increases with absorbed dose. Also, the ultrasonic speed in polymer gel dosimeters is affected by changes in dosimeter elastic modulus that are likely to be a result of polymerization. It is suggested that ultrasound is a sufficiently sensitive technique for polymer gel dosimetry. PMID- 12539981 TI - The use of diode array spectroradiometers for dosimetry in phototherapy. AB - An evaluation of two diode array radiometers, an UV spectroradiometer, Type SC-MP A, from 4D Controls (Redruth, UK) and an USB2000-UV-VIS spectrometer from Ocean Optics (Duiven, NL), was carried out at the Photobiology Unit, University of Dundee. Three parameters of the instruments' performance were investigated, having been identified as the most likely sources of error in phototherapy dosimetry: (1) calibration, (2) stray light rejection, (3) angular response. An assessment was then made of the reliability of this type of instrument for dosimetry in clinical practice by measurement of a selection of phototherapy sources, in direct comparison with calibrated radiometers. Both instruments were found to have significant stray light levels (SC: 13% and USB: 39%). The use of stray light compensation and a high output calibration source improves accuracy to within acceptable limits. Angular responses were satisfactory: f2 values (+/- 60 degrees) of 5.9% and 7.8% for SC and USB, respectively. The SC spectroradiometer is supplied as a calibrated instrument. Using the supplied calibration resulted in errors in measuring phototherapy sources of up to 44% in UVA. Alternative calibration reduced the error in measuring UVA and UVB sources to within 12%. The USB spectrometer was found to have insufficient responsivity in both UVB and UVA to provide reproducible measurements of most phototherapy sources. PMID- 12539982 TI - Pre-clinical evaluation of an inverse planning module for segmental MLC based IMRT delivery. AB - Phantom tests are performed for pre-clinical evaluation of a commercial inverse planning system (HELAX TMS, V 6.0) for segmented multileaf collimator (MLC) intensity modulated radiotherapy (IMRT) delivery. The optimization module has available two optimization algorithms: the target primary feasibility and the weighted feasibility algorithm, only the latter allows the user to specify weights for structures. In the first series, single beam tests are performed to evaluate the outcome of inverse planning in terms of plausibility for the following situations: oblique incidence, presence of inhomogeneities, multiple targets at different depths and multiple targets with different desired doses. Additionally, for these tests a manual plan is made for comparison. In the absence of organs at risk, both the optimization algorithms are found to assign the highest priority to low dose constraints for targets. In the second series, tests resembling clinical relevant configurations (simultaneous boost and concave target with critical organ) are performed with multiple beam arrangements in order to determine the impact of the system's configuration on inverse planning. It is found that the definition of certain segment number and segment size limitations does not largely compromise treatment plans when using multiple beams. On the other hand, these limitations are important for delivery efficiency and dosimetry. For the number of iterations and voxels per volume of interest, standard values in the system's configuration are considered to be sufficient. Additionally, it is demonstrated that precautions must be taken to precisely define treatment goals when using computerized treatment optimization. Similar phantom tests could be used for a direct dosimetric verification of all steps from inverse treatment planning to IMRT delivery. PMID- 12539983 TI - Comparison of TG-43 dose calculations to pinpoint ion chamber and diamond detector measurements. AB - Brachytherapy is an area of radiation therapy where the availability of high resolution detectors with a low energy-dependence is of great importance. The suitability of two detectors is investigated in this study. Measurements in the proximity of an Ir-192 source were performed with a diamond detector type 60003 and a pinpoint chamber type 31006 (both PTW-Freiburg). For comparison, dose values were calculated with the dose calculation formalism recommended by the task group 43 (Nath et al 1995 Med. Phys. 22 209-34). PMID- 12539984 TI - X-ray CT high-density artefact suppression in cryosurgery. AB - Advantages of x-ray CT for imaging guidance of cryosurgery include 3D visualization of frozen and unfrozen tissue and calibration of temperature in the tissue water-ice interface (0-10 degrees C) to Hounsfield units. However, use of x-ray CT images and their thermal calibration can be compromised by the cryoprobes generating high-density streak artefacts. A new subtraction technique for artefact suppression is proposed and tested in prostate cryosurgery simulations. By subtracting the measured CT x-ray projection profile without cryoprobes from the profile with cryoprobes plus iceballs, one obtains the combined profile of the cryoprobes and a low value background. Polynomial interpolation to obtain the background profile allows its addition to the original profile without probes. The result may then be fed to a conventional filtered back-projection routine to reconstruct the probe-free image. Finally the cryoprobe pixels in the originally constructed image with probes and iceballs are added back to the probe-free image to get the final artefact-suppressed image. The major advantage of this subtraction technique is that it can successfully suppress the high-density artefacts in bone-abundant body regions such as the pelvis. X-ray CT images of cryoprobe arrays in a homogeneous gelatin phantom and the pelvic region of an anthropomorphic Rando phantom containing a human skeleton were generated. After suppression, cryoprobe metal artefact streaks are reduced and visualization of the positions and dimensions of the cryoprobes are well preserved. PMID- 12539985 TI - Thyrotoxic periodic paralysis. PMID- 12539986 TI - A new home and a new name: Winston-Salem and the Bowman Gray School of Medicine, 1941-1971. PMID- 12539987 TI - Research centers in arteriosclerosis, stroke, and cancer at Wake Forest University School of Medicine. PMID- 12539988 TI - "Like a star sapphire": reflections on basic science at Wake Forest University School of Medicine. PMID- 12539989 TI - The evolution of public health sciences in an academic medical center. PMID- 12539990 TI - Management of hyperlipidemia in the elderly population: an evidence-based approach. AB - Coronary heart disease (CHD) is the leading cause of morbidity and mortality in patients more than 65 years old. Within this population, elevated cholesterol levels are prevalent and associated with increased risk of CHD. Despite increasing emphasis on lipid-lowering treatment in the elderly population, questions remain regarding secondary and primary prevention of CHD. According to current clinical trial evidence, lipid-lowering therapy, specifically with HMG CoA-reductase inhibitors, can reduce CHD morbidity and mortality without increased adverse effects in the elderly population. Lipid-lowering treatment should be considered for patients aged 65 to 75 years with a history of CHD or who are at moderate to high risk for CHD. Estrogen replacement therapy (ERT), which has also been shown to lower cholesterol levels, raises special considerations for postmenopausal women. However, recent findings suggest that postmenopausal women with a history of CHD should not be given estrogen solely for secondary prevention of CHD events. PMID- 12539991 TI - Hospital presentation after stroke in a community sample: the Mobile Stroke Project. AB - BACKGROUND: Existing data regarding time between stroke and presentation for treatment are largely derived from hospital-based or multicenter databases and may not accurately reflect presentation patterns for most hospitalized stroke patients. METHODS: We evaluated a consecutive series of all hospitalized patients in Mobile County, Alabama. RESULTS: We identified 1,010 hospitalized stroke patients. Of all patients with out-of-hospital stroke, 42% came to a hospital within 3 hours of symptom onset. There were no statistically significant interhospital differences. Being asleep at the time of stroke or being transported by family or friends significantly increased the likelihood of late arrival. CONCLUSIONS: A minority of stroke patients arrive at a hospital early enough to qualify for acute intervention. Until development of acute therapies with longer therapeutic windows or more robust therapeutic benefit than tissue plasminogen activator (t-PA), effective stroke prevention strategies will exert a greater influence on stroke incidence and morbidity. PMID- 12539992 TI - Health care priorities for menopausal women with a history of breast cancer. AB - BACKGROUND: The opinion of breast cancer survivors and their physicians about long-term health, especially menopause, is not well understood. METHODS: Seventy three patients and 22 physicians answered questions regarding medical follow-up and menopause. RESULTS: One third of specialists preferred follow-up of 5 years or less, while 59% preferred 10 years or longer; 46% of patients preferred follow up for 10 years or longer. Physicians preferred that primary care physicians supervise menopausal health (55%), but patients disagreed (30%). Physicians cited heart health most important, followed by skeletal health and climacteric symptoms. Physicians believed that climacteric symptoms were patients' leading concern, but patients cited heart health, followed by skeletal health and cognitive dysfunction. Neither patients nor physicians advocated estrogen use. CONCLUSIONS: Differences of opinion exist between breast cancer patients and specialists regarding follow-up and management of menopause. However, both patients and physicians prefer prolonged surveillance by a cancer specialist, with attention to heart and skeletal health issues. PMID- 12539993 TI - Ursodeoxycholic acid in the treatment of cholestasis and hyperbilirubinemia in pediatric intensive care unit patients. AB - BACKGROUND: The purpose of this study was to evaluate the efficacy of ursodeoxycholic acid (UDCA) in the treatment of cholestasis and hyperbilirubinemia in a pediatric intensive care unit population. METHODS: Medical records were retrospectively reviewed to identify children and adolescents in the pediatric intensive care unit (PICU) who received UDCA for the treatment of hyperbilirubinemia and cholestasis. RESULTS: Ursodeoxycholic acid was administered at a dose of 20 mg/kg per day to 5 PICU patients with cholestasis and hyperbilirubinemia of various etiologies. In 4 of 5 patients, there was a decrease in serum bilirubin levels following the start of UDCA therapy. There was no response to therapy in 1 patient, who developed disseminated fungal disease and died. No adverse effects related to therapy were noted. CONCLUSIONS: These preliminary data suggest that UDCA is effective in the treatment of cholestasis and hyperbilirubinemia of various etiologies in the PICU patient. Prospective, randomized trials are warranted to further assess the efficacy of this therapy in this patient population. PMID- 12539994 TI - Acid-base and electrolyte disturbances in patients with hypercalcemia. AB - BACKGROUND: In the present study, we analyzed acid-base and electrolyte disturbances in hypercalcemic patients to determine the principal causes of hypercalcemia. METHODS: We studied a total of 76 hypercalcemic patients and 91 healthy individuals. Acid-base and electrolyte parameters were determined before any therapeutic intervention. RESULTS: Hyperparathyroidism and neoplasias were the most common causes of hypercalcemia. Hypercalcemic patients had increased serum urea and creatinine levels, a higher urea/creatinine ratio, and a higher rate of acid-base disorders, but lower serum albumin, potassium, chloride, phosphorus, and magnesium concentrations than those found in the control subjects. Notably, significant differences in acid-base balance and electrolyte concentrations were evident between patients with hyperparathyroidism and patients with cancer. CONCLUSIONS: Primary hyperparathyroidism and neoplasia are the most common causes of hypercalcemia. A wide array of concurrent acid-base and electrolyte disorders may be evident in hypercalcemic patients. Differences in these laboratory parameters are helpful in diagnostic workup. PMID- 12539995 TI - Epidemiology of fatty liver in a hospital-based study in Taiwan. AB - BACKGROUND: The aim of this study was to assess the epidemiology of fatty liver in Taiwan. METHODS: We retrospectively analyzed the medical records of all patients receiving health examinations at China Medical College Hospital from January through December 2000. A total of 1,012 subjects were included for analysis. Data were analyzed using the t test, chi-square analysis, and multivariate logistic regression. RESULTS: Our cohort was comprised of 41.8% women and 58.2% men; mean age was 49.2 years. The prevalence of fatty liver was 36.9% in all subjects, and prevalence was higher in men than in women. After controlling for other covariates, the multivariate logistic regression analysis showed that the significant factors related to fatty liver were male sex, obesity, hyperglycemia, hypertriglyceridemia, elevated glutamate pyruvate transaminase level, and hyperuricemia. Elevated glutamate oxaloacetate transaminase level was negatively associated with fatty liver. CONCLUSIONS: The prevalence of fatty liver is high in Taiwan. This study establishes background data for further investigation. PMID- 12539996 TI - Pneumococcal susceptibility to meropenem in a mid-south children's hospital. AB - BACKGROUND: We investigated pneumococcal susceptibility to meropenem in isolates from a tertiary children's hospital where pneumococci are commonly resistant to penicillin and cefotaxime. METHODS: From July 1998 to August 1999, meropenem susceptibilities were determined by E-test for all Streptococcus pneumoniae isolates from blood or cerebrospinal fluid and for penicillin-nonsusceptible pneumococcal isolates from other sites. RESULTS: Isolates that were penicillin susceptible or penicillin-intermediate were all susceptible to meropenem. Of 29 penicillin-resistant isolates, 27 were nonsusceptible to meropenem (13 intermediate, 14 resistant). Cefotaxime-susceptible isolates were all susceptible to meropenem. Of 11 cefotaxime-intermediate isolates, 10 were nonsusceptible to meropenem (9 intermediate, 1 resistant). Of 20 cefotaxime-resistant isolates, 17 were nonsusceptible to meropenem (4 intermediate, 13 resistant). CONCLUSIONS: Meropenem resistance is common among pneumococci with decreased susceptibility to penicillin or cefotaxime. The role of this agent in the treatment of invasive infections caused by pneumococci that are resistant to penicillin and cefotaxime may be limited. PMID- 12539997 TI - Hospitalizations and outcomes for diabetic gastroparesis in North Carolina. AB - BACKGROUND: Population-based data to assess the extent of gastroparesis in the diabetic population are scarce. We examined the demographic and clinical characteristics associated with hospital admissions for diabetic gastroparesis in North Carolina. METHODS: Data from the 1998 North Carolina Hospital Discharge database were abstracted from records in which gastroparesis and diabetes mellitus were listed as simultaneous diagnoses. RESULTS: There were 1476 discharges meeting our criteria, with total charges of $11,378,446 over 7850 total hospital days. Most patients were female (65.8%), > or = 45 years of age (54.5%), and had Medicare as the primary payer (52.1%). While most of these patients were admitted under emergency or urgent circumstances, the vast majority had routine discharges. CONCLUSION: Despite some limitations, these data indicate that diabetic gastroparesis is not uncommon, but can be treated effectively. PMID- 12539998 TI - Epidemiology of fatal occupational injuries in Jefferson County, Alabama. AB - BACKGROUND: In Alabama, little is known about the overall characteristics of job related injuries. METHODS: We examined fatal occupational injuries that occurred in Jefferson County, Alabama, from 1990 through 1999 using files maintained by the Jefferson County Coroner/Medical Examiner Office. Mortality rates were calculated by sex, age, race, occupation, and industry. RESULTS: There were 186 fatal occupational injuries; 136 were unintentional. The overall fatality rate was 4.45 deaths per 100,000 worker-years. Mortality rates were higher among men than among women, and they increased with advancing age. The homicide rate was higher among black workers than among whites. Mining, manufacturing, and transportation were the industries with the highest fatal injury rates. Transportation-related jobs and manual labor were high-risk occupations. CONCLUSION: High-risk industries should consider worker-targeted injury prevention programs, and particular attention should be paid to elderly workers. PMID- 12539999 TI - Primary human immunodeficiency virus. AB - The term "primary HIV infection" refers to the period from initial infection with the human immunodeficiency virus to complete seroconversion. It is a period of extreme infectiousness. The occurrence and severity of symptoms during primary HIV infection correlate with the rapidity of clinical and immunologic decline. Treatment of patients during primary infection may improve immune preservation and reconstitution. In this review article, we present information that will help clinicians understand, recognize, and diagnose primary HIV infection. The current approach to management of primary HIV infection is based more on expert opinion than clinical trial results, though ongoing clinical trials should provide more information about this syndrome. PMID- 12540000 TI - Effect of a patient care partnership project on cost and quality of care at an academic teaching hospital. AB - BACKGROUND: In 1998, the Patient Care Partnership Project was conducted by general internal medicine physicians and hospital administration in an academic health care center. The project was designed to optimize cost, quality, and service results to inpatients. METHODS: The project focused on improved communication among physicians, a nurse discharge planner, and hospital administration regarding appropriate resource utilization. The outcomes were average cost per inpatient, length of hospital stay, 30-day readmission rates, mortality rates, and resident and patient satisfaction. Comparisons were made with three control groups. RESULTS: The postintervention generalist-staffed services showed significant reductions in average costs per patient and length of stay. These parameters increased in the specialist group from 1997 to 1998. Readmission rates remained stable, and mortality rates actually decreased. Patient and resident satisfaction remained unchanged. CONCLUSIONS: A collaborative effort between generalists and hospital administration led to a significant improvement in resource utilization compared with the three control groups, with no compromise in quality outcomes. PMID- 12540001 TI - Amiodarone-induced thyrotoxicosis presenting as hypokalemic periodic paralysis. AB - Thyroid disorder is a well-recognized side effect of amiodarone therapy. Thyrotoxicosis is less common than hypothyroidism. Hypokalemic periodic paralysis is one manifestation of thyrotoxicosis, and is more often seen in Oriental and Latin American men than in other demographic groups. This phenomenon, however, has not been previously described in thyrotoxicosis due to amiodarone usage. We describe a case of amiodarone-induced thyrotoxicosis in a 34-year-old man who presented with sudden lower extremity weakness, heat intolerance, and weight loss. Physical examination demonstrated fine tremors. Serum potassium level was 2.2 mEq/L on admission. Gastrointestinal and renal causes of potassium loss were excluded by history and physical examination. Further biochemical testing demonstrated abnormal thyroid function. The urinary potassium and serum bicarbonate, magnesium, and calcium levels were within normal limits. Lower extremity weakness resolved immediately after potassium replacement therapy. Methimazole therapy was initiated, and the patient was clinically euthyroid on discharge. PMID- 12540002 TI - Pneumonia due to Mycoplasma pneumoniae with transient proteinuria. AB - Mycoplasma pneumoniae pneumonia can be associated with several systemic features. This report illustrates a case of M. pneumoniae pneumonia in an adult who recovered completely. Direct infection of the kidneys or an immunologic mechanism may have been the underlying cause. This patient also had simultaneous pharyngitis caused by group A beta-hemolytic streptococci. Various clinical possibilities with respect to pneumonia, pharyngitis, and proteinuria are briefly reviewed. PMID- 12540003 TI - Lightning injuries. AB - Lightning strikes may cause a constellation of injuries. Blunt head trauma, neurologic injury, and cardiac injury are common in these patients. In contrast to high-voltage electrocutions, blunt trauma after a lightning strike is common. Thorough evaluation of all organ systems is crucial. This report discusses mechanism of injury and describes initial evaluation and treatment of lightning strike victims. PMID- 12540004 TI - Association of breast cancer and immune thrombocytopenic purpura. AB - Immune thrombocytopenic purpura (ITP) and breast cancer are common disorders. Only six cases in which patients have had both diseases have been reported. We describe a 40-year-old woman who had ITP while responding to therapy for metastatic breast cancer. Given the few reported cases, the diverse presentations of thrombocytopenia during the course of each patient's breast cancer and the variable therapeutic responses of ITP, the association of breast cancer and ITP is probably coincidental. PMID- 12540006 TI - Spontaneous intracranial hypotension. AB - Spontaneous intracranial hypotension (SIH) is an increasingly recognized syndrome. Postural headache with typical findings on magnetic resonance imaging (MRI) are the key to diagnosis. Delay in diagnosing this condition may subject patients to unnecessary procedures and prolong morbidity. We describe a patient with SIH and outline the important clinical and radiographic features of this syndrome. Headache due to SIH is similar to headache occurring after lumbar puncture. Patients with postural headaches should have brain MRI before lumbar puncture. Meningeal abnormalities with typical clinical features are helpful in establishing the diagnosis. When correctly diagnosed, SIH management, in most cases, is easy and highly effective. PMID- 12540005 TI - Gastrointestinal bleeding in adult patients with Meckel's diverticulum: the role of technetium 99m pertechnetate scan. AB - Obscure gastrointestinal (GI) bleeding is often challenging for the primary care physician, but with improved diagnostic testing the cause of this blood loss is determined in most patients. However, approximately 5% of the time no underlying cause is found. One common etiology in patients younger than 40 years of age is a Meckel's diverticulum. The technetium 99m pertechnetate scan is the standard test for making this diagnosis. However, the sensitivity of the scan is only 62% in the adult population. In this case report, a patient with profound, hemodynamically significant GI blood loss had multiple negative studies. Subsequently, an abnormal vascular lesion was detected and during exploratory laparotomy, a Meckel's diverticulum was found and removed. Although the technetium pertechnetate scan is falsely negative in a number of cases, there are ways to increase its sensitivity and possibly avoid repeated testing. PMID- 12540007 TI - Hypothyroidism and muscular respiratory failure successfully treated with liothyronine. AB - After total thyroidectomy because of hyperthyroidism, hypothyroidism developed in a 78-year-old woman. Despite replacement therapy with levothyroxine sodium in continuously increasing doses, worsening hypothyroidism led to respiratory failure, necessitating artificial ventilation. The addition of liothyronine resulted in complete recovery. Impaired conversion of thyroxine to triiodothyronine by the dejodase was responsible for the manifestations of hypothyroidism. PMID- 12540008 TI - Intra-abdominal abscess caused by Listeria monocytogenes in a patient with acquired hemolytic anemia and thrombocytopenia. AB - We report a case of an intra-abdominal abscess caused by Listeria monocytogenes in a postoperative patient with Evans syndrome (acquired hemolytic anemia and thrombocytopenia). Focal infections with L. monocytogenes are uncommon but have been reported in immunocompromised patients. A few cases of liver abscess in diabetic patients have also been reported. The present case is significant because of the paucity of previously described focal intra-abdominal infections caused by L. monocytogenes, particularly as a postsplenectomy pathogen. PMID- 12540009 TI - Doxycycline-induced hypoglycemia in a nondiabetic young man. AB - The medication history is an integral part in the evaluation of a patient with hypoglycemia. A variety of medications have been associated with hypoglycemia, but the list of these medications is expanding. We report the first case of doxycycline-induced hypoglycemia in a young nondiabetic man. PMID- 12540010 TI - Behcet's syndrome coexisting with clinically occult ankylosing spondylitis. PMID- 12540011 TI - Rhabdomyolysis of infectious and noninfectious causes. PMID- 12540012 TI - Into the future: genetic testing for type 2 diabetes mellitus. PMID- 12540013 TI - An unusual progression of untreated osteosarcoma. PMID- 12540014 TI - Symptom improvement in patients with onychomycosis receiving oral antifungal therapy. PMID- 12540015 TI - Shedding light on vascular permeability during peritonitis: role of mast cell histamine versus macrophage cysteinyl leukotrienes. AB - The inflammatory response consists of sequential steps that are essentially the same whatever the cause and wherever the site. The main purpose of inflammation is to bring fluid, proteins, and cells from the blood into the damaged tissues. Therefore there are mechanisms that allow cells and proteins to gain access to extravascular sites, where and when they are needed if damage and infection has occurred. A critical process for formation of inflammatory exudate is an increase in permeability of local blood vessels. Vasopermeability changes can be usually attributed to mast cells and their mediators but recent studies reveal that also macrophages can be involved in this process. This short commentary discusses new data on cellular origin of major vasoactive mediators, and their receptors during peritoneal inflammation in mice. PMID- 12540016 TI - RDP58, a locally active TNF inhibitor, is effective in the dextran sulphate mouse model of chronic colitis. AB - OBJECTIVE AND DESIGN: RDP58 is a novel anti-inflammatory peptide that inhibits TNF synthesis and upregulates heme oxygenase-1. RDP58 therapy was evaluated in the dextran sodium sulphate (DSS) model of chronic colitis. MATERIAL: Colitis was induced by giving DSS to mice (n = 8 animals/group). Toxicity studies were done in Rhesus monkeys (n = 5), dogs (n = 3) and mice (n = 10). TREATMENT: In colitis, mice were treated with p.o. vehicle (saline), RDP58 (5 and 10 mg/kg/day) or 5-ASA (50 mg/kg/day). METHODS: Disease activity index (DAI) was used as the endpoint of efficacy. RESULTS: RDP58 therapy significantly reduced DAI and histological scores in all animals. DAI scores in RDP58 treated animals declined faster than 5 ASA. RDP58 at 5 or 10 mg/ kg/day significantly reduced DAI compared to 5-ASA. RDP58 significantly reduced acute, chronic and total inflammation scores. It enhanced re-epithelialization by reducing crypt scores. RDP58 was not bioavailable and was well tolerated. CONCLUSIONS: Therapeutic efficacy of RDP58 combined with a lack of bioavailibility and toxicity suggest that RDP58 may be a promising new therapeutic for IBD. PMID- 12540017 TI - Lower apoptosis rate and higher CD69 expression in neutrophils from atopic individuals. AB - OBJECTIVE: In this study we aimed to examine the kinetics of CD69 expression and the susceptibility to apoptosis, in eosinophils and neutrophils, in the presence or absence of GM-CSF. We also addressed the question whether differences between atopic patients and healthy individuals exist in this respect. MATERIALS AND METHODS: Freshly isolated eosinophils and neutrophils from non-atopic and atopic donors were analysed by flow cytometry for Annexin/PI staining, caspase 3 activation and CD69 expression. RESULTS: We found a higher CD69 expression when atopic neutrophils were incubated with GM-CSF compared to non-atopic neutrophils, and that the kinetics of CD69-expression in neutrophils, but not in eosinophils, differed between non-atopic and atopic individuals (p < 0.004). We also found a higher viability in GM-CSF-stimulated neutrophils from non-atopic individuals as compared to neutrophils from atopic individuals (p < 0.05). CONCLUSION: These data suggest a potential role for neutrophils in the allergic inflammatory reaction through differences in apoptosis rates and CD69 expression between atopic and non-atopic individuals. PMID- 12540018 TI - Inhibition of antigen-induced arthritis in guinea pigs by a selective LTB4 receptor antagonist LY293111Na. AB - OBJECTIVE AND DESIGN: To investigate the role of leukotriene B4 (LTB4) and prostaglandin E2 (PGE2) in the development and progression of antigen-induced arthritis (AIA) in guinea pigs and rats. METHODS: Arthritis was induced by injecting cationic amidated bovine serum albumin (aBSA) into the knee joint of immunized guinea pigs or rats. The effect of a potent and selective LTB4 receptor antagonist, LY29311INa (2-[2-propyl-3-[3-[2-ethyl-4-(4-fluorophenyl)-5 hydroxyphenoxy]-propoxy]-phenoxy]benzoic acid sodium salt) was compared with those of indomethacin and dexamethasone. The effect of LY293111Na on adjuvant arthritis in rats was also examined. RESULTS: LY293 111Na (5 to 50 mg/kg b.i.d.) significantly inhibited knee joint swelling and histopathological changes of AIA in guinea pigs, but not in rats. Especially its protective effect against bone and cartilage destruction was substantial. In contrast, the cyclooxygenase inhibitor indomethacin significantly inhibited AIA in rats, but slightly inhibited in guinea pigs, while dexamethasone markedly inhibited AIA in both guinea pigs and rats. Increases of LTB4 and myeloperoxidase (MPO) activity were observed in the knee joint tissue of AIA guinea pigs, and LY293111Na dose dependently inhibited the increase of MPO activity. Moreover, in adjuvant arthritic rats, LY293111Na showed slight inhibitory effect, while indomethacin showed marked inhibition. CONCLUSIONS: LTB4 but not PGE2 appeared to play important roles as an effective mediator in joint, particularly in cartilage and bone destruction of AIA in guinea pigs probably by inducing polymorphonuclear leukocytes (PMNs) chemotaxis to the joint tissue. In contrast, PGE2 but not LTB4 is an important mediator of arthritis in rats. PMID- 12540019 TI - Endogenous central histamine-induced reversal of critical haemorrhagic hypotension in rats--studies with histamine N-methyltransferase inhibitor SKF 91488. AB - OBJECTIVE AND DESIGN: Activation of the histaminergic system is associated with mobilisation of compensatory mechanisms in response to the action of stimuli which disturb homeostasis. Therefore, the effects of endogenous histamine in central cardiovascular regulation were studied in a rat model of irreversible haemorrhagic hypotension. MATERIAL: Cardiovascular parameters were measured in 66 and central histamine concentrations in 12 male Wistar rats anaesthetised with ketamine/xylazine. TREATMENT: Haemorrhage-shocked rats with mean arterial pressure (MAP) 20-25 mmHg were injected intracerebroventricularly (icv) with histamine N-methyltransferase inhibitor SKF 91488 after icv pre-treatment with H, H2 and H3 histamine receptor antagonists--chlorpheniramine (100 nmol), ranitidine (200 nmol) and thioperamide (100 nmol), respectively, or saline. METHODS: Arterial pressure and heart rate (HR) changes were monitored within 2 h after icv treatment, or to death if it occurred earlier. Histamine concentrations were measured using enzyme immunoassay. ANOVA followed by a test of Neuman-Keules, and Fisher's exact test were used to compare the results. RESULTS: SKF 91488 produced dose-dependent, significantly higher compared to normovolaemic animals, increases in MAP and HR with the improvement in survival rates. The action of SKF 91488 (100 microg) was associated with an increase in endogenous histamine concentrations in the cerebral cortex (1.19 +/- 0.09 vs. 0.77 +/- 0.21 nmol/g; p < 0.05), hypothalamus (5.62 +/- 0.68 vs. 4.18 +/- 0.45 nmol/g; p < 0.01) and medulla oblongata (0.53 +/- 0.17 vs. 0.29 +/- 0.05 nmol/g; p < 0.05) in comparison to saline-treated animals. SKF 91488-induced effects were inhibited by chlorpheniramine, whereas neither ranitidine nor thioperamide influenced the action. CONCLUSION: Endogenous central histamine, after SKF 91488 treatment, via activation of H, receptors produces reversal of hypotension, with improvement in the survival rate at 2 h after treatment, in rats subjected to critical haemorrhagic hypovolaemia. PMID- 12540020 TI - Effect of H1-antagonist Dithiaden on human PMN-leukocyte aggregation and chemiluminescence is stimulus-dependent. AB - OBJECTIVE AND DESIGN: Contradictory data published on histamine-PMN leukocyte interactions stimulated us to study to the role of histamine and H1-antagonist Dithiaden in generation of reactive oxygen species (ROS) and aggregation of human neutrophils. METHODS AND MATERIALS: Whole blood or isolated PMN-leukocytes were exposed in a dose-dependent way to histamine or H1-antagonist Dithiaden and subsequently stimulated. Whole blood was stimulated with opsonised zymosan (OZ). Isolated cells were stimulated with membrane stimuli (OZ, N-formyl-methionyl leucyl-phenylalanine--fMLP), or membrane bypassing stimuli (Ca2+-ionophore A23187, phorbol-myristate-acetate--PMA). The luminol-enhanced chemiluminescence (CL) was measured separately (whole blood) in a luminometer or simultaneously with neutrophil aggregation in a whole blood lumiaggregometer. RESULTS: Depending on the concentration used, Dithiaden" was 1.5- to 25.0-times more effective in inhibiting activated CL of whole blood than histamine. In isolated neutrophils both histamine and Dithiaden inhibited OZ- and A23187-stimulated CL dose dependently, with potentiation observed after stimulation with PMA and fMLP. Histamine did not alter aggregation with any of the stimuli tested. Dithiaden inhibited A23187-, OZ- and PMA-stimulated PMN-leukocytes but potentiated fMLP induced aggregation of isolated neutrophils. Simultaneous application of Dithiaden and histamine abolished the effect of Dithiaden on fMLP-stimulated CL. CONCLUSIONS: Dithiaden, depending on the stimuli applied, inhibited human neutrophils, both isolated or in whole blood, more markedly than histamine. The inhibition of aggregation and CL was dose- and stimulus-dependent. Histamine administered simultaneously abolished the effect of Dithiaden on fMLP-stimulated PMN-leukocytes. It seems likely that the interaction of Dithiaden with neutrophils operated both at an extra- and intracellular level. PMID- 12540021 TI - Elevated plasma levels of vascular endothelial growth factor and plasminogen activator inhibitor-1 decrease during improvement of psoriasis. AB - OBJECTIVE AND DESIGN: An evaluation of angiogenesis related molecules during open treatment of psoriasis. MATERIALS AND SUBJECTS: Plasma samples and skin biopsies from 16 patients with psoriasis and plasma samples from 13 healthy controls. TREATMENT: Ranitidine 300 mg orally twice daily for 6 months. METHODS: Vascular endothelial growth factor (VEGF) and plasminogen activator inhibitor-1 (PAI-1) were determined by ELISA methods in plasma collected from the patients before treatment and after 1, 3 and 6 months. Vessel counts were performed in biopsies from affected skin areas taken before treatment and after 3 and 6 months. The results were compared to simultaneous PASI scores. RESULTS: Pre-treatment plasma levels of VEGF and PAI-1 were significantly elevated in patients compared with levels in healthy persons (p = 0.02 and p = 0.04, respectively). The plasma levels decreased significantly during treatment (p = 0.03 and p = 0.01, respectively), and the decrease in combined levels correlated with the decrease in PASI score. However, the vessel density in affected skin did not change during treatment. CONCLUSIONS: Increased pre-treatment levels of VEGF and PAI-1 and decrease during improvement of the disease suggest that the two molecules may play a role in pathogenesis of psoriasis. PMID- 12540022 TI - The methylenetetrahydrofolate reductase (MTHFR) gene in colorectal cancer: role in tumor development and significance of allelic loss in tumor progression. AB - BACKGROUND AND AIM: Folate deficiency predisposes to sporadic colorectal cancer (CRC). Methylenetetrahydrofolate reductase (MTHFR) is a critical folate metabolising enzyme and a polymorphism at position 677 (C677T), is associated with reduced enzyme activity. We investigated whether this functional polymorphism modulates the risk of developing CRC. METHODS: This was a retrospective case-control study. 136 unselected cases of sporadic CRC and 848 normal population controls were genotyped for the MTHFR C677T polymorphism. Tumor tissue was genotyped to assess loss of heterozygosity (LOH). RESULTS: MTHFR CT heterozygotes had a significantly increased risk of developing CRC (53.7% of CRC cases vs 38.4% of controls), odds ratio 1.86 (95% CI 1.3-2.7, p < 0.005). No increased cancer risk was observed in TT homozygotes. The MTHFR 'T' allele frequency was significantly higher in the cancer group (0.3713) as compared to controls (0.2900, p < 0.008). LOH at the MTHFR locus was observed in 18% of informative cancers, with exclusive loss of the variant 'T' allele, in all cases. CONCLUSION: In this study of a homogenous northern European population, MTHFR CT heterozygotes had an almost two-fold increased risk of developing sporadic CRC. The exclusive pattern of MTHFR allele loss in cases of LOH, suggest that functional MTHFR activity within a tumor might play an important role in the survival and progression of a colonic neoplasm. PMID- 12540023 TI - Expression of proteinase-activated receptor 2 during taurocholate-induced acute pancreatic lesion development in Wistar rats. AB - BACKGROUND: Proteinase-activated receptor 2 (PAR-2) is a G-protein coupled transmembrane receptor activated by trypsin by site-specific cleavage. Its presence on pancreatic structures was demonstrated in the past. PAR-2 physiologically involves in duct/acinary cells secretion, arterial tonus regulation or capillary liquid turnover. During development of acute pancreatitis/acute pancreatic lesion (APL) these mentioned structures are influenced by very high concentration of trypsin due to its increased basolateral secretion into the interstitium. The aim of our study as presented was to investigate whether PAR-2 is also involved in APL following changes of PAR-2 expression. METHODS: APL was investigated in Wistar rats after injection of 0.1 mL taurocholate into the ductus choledochus. Anatomy, histology, reverse transcriptase polymerase chain reaction (RT PCR) as well as immunohistochemistry and Western-blot analysis of pancreatic tissue were performed using antibody mapping of the new NH2 terminal of PAR-2 after trypsin cleavage. Results from control rats and d 1 or d 4 rats after taurocholate injection were compared. RESULTS: Much higher positivity on acinary/duct cells was observed in APL induced animals than in controls. Similar findings were noticed on arterial smooth muscle cells. Surprisingly, parallel to the exocrine pancreas and vessel findings, enhanced Langerhans' islet cell positivity was observed in experimental animals. CONCLUSIONS: Based on these results, we have demonstrated that during APL development PAR-2 expression increases. This effect is caused by conformational changes after PAR-2 activation, and the new NH2 terminal of activated receptor presentation. We suggest that PAR-2 physiological functions are enhanced during APL development. PMID- 12540024 TI - Phase I study of concomitant gemcitabine and IMRT for patients with unresectable adenocarcinoma of the pancreatic head. AB - PURPOSE: We hypothesized that dynamic intensity-modulated radiotherapy (IMRT) would protect normal tissues enough to allow the escalation of either the gemcitabine or radiotherapy dose in unresectable pancreatic cancer patients. METHODS AND MATERIALS: The trial was designed to build on a previous phase I trial that determined the maximum tolerated dose (MTD) of gemcitabine (350 mg/m2) with concurrent radiotherapy (30 Gy/10 fractions). Only patients with unresectable disease based on established criteria were eligible. The plan was to alternate escalating the radiation dose by 3 Gy and the gemcitabine dose by 50 mg/m2. The starting dose of gemcitabine was 350 mg/m2 and 33 Gy/11 fractions of IMRT to the regional lymphatics and primary disease. The NCI Common Toxicity Criteria were used for dose-limiting toxicity (DLT). RESULTS: All three patients in the first cohort treated suffered DLT. Therefore, a second cohort of patients received a lower gemcitabine dose (250 mg/m2). Both patients treated at this dose level experienced DLT. The DLTs were all due to myelosuppression and upper gastrointestinal toxicity. All patients required a gemcitabine dose reduction. Also, four patients required hospital admission for supportive care, while the fifth died of an unrelated cause shortly after completing therapy. The trial was then closed due to excessive toxicity. CONCLUSION: Hypofractionated dynamic IMRT to the primary site and regional lymphatics did not permit escalation of either the radiation or gemcitabine dose. Dynamic IMRT requires further investigation before it can be applied to toxic combinations of chemotherapy and radiation in the upper abdomen. PMID- 12540025 TI - Validation of virtual colonoscopy in the detection of colorectal polyps and masses: rationale for proper study design. AB - BACKGROUND: Colorectal cancer, the second-leading cause of cancer-related mortality, is a preventable malignancy in many cases. Despite the availability of several screening modalities, compliance with screening recommendations remains unacceptably low. Virtual colonoscopy is a novel, minimally-invasive technique with the potential to increase colorectal cancer screening rates, but its effectiveness must first be validated. Published studies comparing virtual colonoscopy to conventional colonoscopy have reported varying results. These discrepancies may be attributed to differences in bowel preparation and scanning techniques, as well as errors in endoscopic lesion measurement, endoscopic colonic segmental localization, and the ability of conventional colonoscopy to actually detect lesions. These methodological issues can affect scientific results and ultimately affect the public's perception of this emerging technique. AIM: The goal of this report is to expose existing methodological shortcomings and propose solutions incorporated in this study design. This article describes the rationale, study design, and outcome definitions of a single-center, blinded, direct comparative trial aiming at assessing the ability of virtual colonoscopy to detect colorectal polyps and masses relative to the criterion standard, conventional colonoscopy. DESIGN FEATURES: Bowel preparation was standardized using oral sodium phosphate lavage, orally administered iodinated contrast, and controlled colonic insufflation. Segmental unblinding allowed a second-look when results were discrepant and polyp matching was performed using an algorithm based on segmental localization and lesion size determination. CONCLUSIONS: This methodology could be applied to other studies assessing the accuracy of virtual colonoscopy in order to have uniformity of results. PMID- 12540026 TI - Surgical management of benign tumors of the liver. AB - INTRODUCTION: Benign tumors of the liver are increasingly being diagnosed and continue to represent a management challenge. These lesions constitute a substantial component of hepatic neoplasms evaluated and resected at a tertiary referral center. We reviewed our experience with resection of benign liver lesions to clarify the safety and effectiveness of this treatment. METHODS: Between January 1996 and January 2000, 28 patients with benign hepatic lesions were identified from a cohort of 140 hepatic resection patients. Demographic characteristics, operative management, morbidity, mortality and follow-up were retrospectively analyzed. RESULTS: The mean age in our patients was 35 +/- 14, with 24/28 (86%) patients being female. Seven of the 24 woman (29%) at presentation were either pregnant or immediate postpartum. A history of OCP use was noted in 14/24 (58%) female patients. The most common presenting symptom was abdominal pain in 12/28 (43%). Resection for an undiagnosed mass occurred in 11/28 (39%) patients. The distribution of pathology was hemangioma 10/28 (35.7%), adenoma 8/28 (28.6%), hepatic cyst 5/28 (17.9%), hamartoma 2/28 (7.1%), and FNH 3/28 (10.7%). Average size of the tumor was 7.4 +/- 3.9 (range 2.5-15 cm) with a mean of 1.4 +/- 0.8 lesions (range 1-3) per patient. Tumors were evenly distributed between the right and left side while eight patients (29%) had bilobar tumors. Enucleation rather than anatomic resection was performed in 18/28 (64%) patients, with a mean blood loss of 457 +/- 532 cc (range 50-2200 cc). Blood transfusion was required in only 3/28 (10%) patients, while total vascular isolation was used in only a single patient undergoing an extended left hepatectomy. Mean length of stay was 6.8 +/- 3.2 d (range 3-14 d). Three complications (10.7%) were encountered: pulmonary embolus, ileus and non operative bile leak. There were no mortalities in this series. Recurrence of tumor occurred in only one patient with a giant hepatic cyst managed laparoscopically. CONCLUSIONS: In our institution, the management of clinically relevant benign tumors of the liver comprises a significant proportion of our resectional practice (20%). Our data suggests that both enucleation and anatomically based resections can be performed safely with minimal blood loss and transfusion requirements. Resection of symptomatic lesions was highly effective in treating abdominal pain due to these benign tumors. We advocate resection of non-resolving hepatic adenomas, symptomatic lesions, or when malignancy cannot be excluded. PMID- 12540028 TI - Iatrogenic esophagobronchial fistula arising in irradiated Barrett's esophagus. AB - A 47-yr-old male underwent a right upper lobectomy for stage IIB bronchoalveolar carcinoma followed by 4600 Gy of irradiation. One year later a fistula formed from an ulcerated region of Barrett's esophagus into the left main bronchus. Bronchotomy repair with onlay patch intercostal muscle flap and esophageal repair with serratus anterior muscle flap plus postoperative esophageal stent placement for stricture resulted in good functional results. PMID- 12540027 TI - Complications of hepatic artery infusion: a review of 4580 reported cases. AB - PURPOSE: The resurgence of hepatic artery infusion (HAI) for the treatment of colorectal liver metastases has been dampened by concern over its complications. We have reviewed the incidence and frequency of complications associated with HAI and discussed the factors associated with these complications. METHODS: A PUBMED search was conducted from 1950-2001 using various combinations of these keywords: hepatic arterial infusion, colorectal carcinoma, complications, and trials. The main inclusion criterion was the reporting of HAI complications. The main exclusion criterion was duplicated patients. Extracted data included chemotherapeutic agents, catheter technique, drug toxicities, and catheter related complications. Relative risks and 95% confidence intervals were calculated. RESULTS: We reviewed 437 articles/abstracts and included 101 studies. 4580 patients with 4582 toxicities and complications were reported. The mortality rate was 1%. The most common toxicities were: GI symptoms 22%, chemical hepatitis 19%, and bone marrow toxicity 8%. 5-fluorouracil (5-FU) HAI had statistically significant risk for GI symptoms and bone marrow toxicity. Floxuridine (FUDR) HAI had statistically significant risk for chemical hepatitis, sclerosing cholangitis, and biliary toxicity. The most common catheter complications were: hepatic artery occlusion 6%, catheter thrombosis 5%, and catheter displacement 7%. CONCLUSIONS: This literature review of the complications of HAI confirms a low mortality associated with HAI. Sclerosing cholangitis and chemical hepatitis are associated with the use of FUDR, while the use of 5-FU is associated with bone marrow toxicity. Our observations support the development of hepatic cytoprotective agents and other effective anti-tumor agents to improve the results and morbidity of HAI for colorectal liver metastases. PMID- 12540029 TI - Solitary true cyst of the pancreas in an adult: report of a case. AB - BACKGROUND: Solitary true cyst of the pancreas is rare in adults, and the differential diagnosis of cystic lesions of the pancreas is challenging. AIM OF THE STUDY: To describe a solitary true cyst of the pancreas in an adult and discuss the differential diagnosis. METHODS: A 50 yr old woman presented with a mass lesion in the right upper quadrant of the abdomen. Abdominal computed tomography showed a cystic lesion, with a maximum diameter of 12 cm, between the inferior surface of the liver and the ascending colon. The cyst was homogenous and had smooth edges. On magnetic resonance imaging, a unilocular cyst was seen that was low intensity on T1-weighted images and very high intensity on T2 weighted images. No connection between the cyst and the pancreatic ductal system was demonstrated on endoscopic retrograde cholangiopancreatography. Laparotomy was performed with a presumptive diagnosis of cystic tumor of the pancreas. RESULTS: On pathologic examination, the cyst was serous and was lined with a single layer of normal cuboidal epithelium that was periodic acid Schiff stain negative. Meticulous examination failed to identify honeycomb-like microcysts characteristic of serous cystadenoma. The final diagnosis was a solitary true cyst of the pancreas. CONCLUSION: The differential diagnosis of solitary true cyst from other cystic lesion of the pancreas usually is based on histology. Particular care must be taken to distinguish this lesion from macrocystic serous cystadenoma. PMID- 12540030 TI - End-stage chronic pancreatitis: a practical disease-descriptor. AB - BACKGROUND: Contemporary classification systems for chronic pancreatitis do not incorporate current knowledge of the disease biology of this condition. METHOD AND RESULTS: This paper presents three case summaries of patients with longstanding chronic pancreatitis with extra-pancreatic complications. The cases highlight the inadequacies of current terminology. The term "end-stage chronic pancreatitis" is applied to advanced disease and disease descriptors are defined. CONCLUSION: The term "end-stage chronic pancreatitis" is a practical and readily applicable disease descriptor. PMID- 12540031 TI - Overview of the 2002 annual scientific conference of the Lustgarten Foundation for Pancreatic Cancer Research: "Pancreatic cancer: translating discovery to patient care". PMID- 12540032 TI - The chelating treatment is not useful in human's intoxication with acute herbicidal organic arsenic. AB - The clinical manifestations of acute organic arsenic intoxication in humans have seldom been described and the associated treatment has been thought to be the same as that of acute inorganic arsenic intoxication. We have studied a collection of patients from 1996 to 2001 who called the Poison Control Center of Kaohsiung Medical University Hospital asking for information regarding acute organic arsenic intoxication. The 17 patients ranged in age from 23 to 64 years old, with 5 females and 12 males. The cause of arsenic ingestion was attempted suicide. Abdominal pain and vomiting were the main symptoms. There were no differences in results between patients treated with and those treated without chelating agents. We therefore believe that the results of acute organic intoxication are not same as acute inorganic intoxication and it is unnecessary to use chelating agents in such conditions. PMID- 12540034 TI - The cytotoxic effects of a traditional Zulu remedy, impila (Callilepis laureola). AB - The traditional Zulu remedy impila (Callilepis laureola) can cause acute fatal hepatocellular necrosis, especially in children. We investigated the mechanism(s) of toxicity using HuH-7 hepatocytes. Impila tubers were extracted with boiling water and the aqueous extract was used at different concentrations to study the effects on the morphology of the cells. Flow cytometry and labelling with fluorescent antibodies to tubulin were also used. At high concentrations, necrosis occurred; however, at lower concentrations, the extracts gave rise to a variety of changes including hypercondensation of chromatin, multinucleate cells, nuclear fragmentation and apoptosis. In addition, we observed destruction of cytoplasmic tubulin. These findings give further insight into the mechanism of toxicity of herbal remedies containing atractyloside. PMID- 12540033 TI - Cytotoxicity in ciprofloxacin-treated human fibroblast cells and protection by vitamin E. AB - Quinolones (Qs) were shown to have cytotoxic effects in various cell lines including human carcinoma cells; however, mechanism of these effects was not fully understood. To investigate the possibility of the involvement of an oxidative stress induction in this mechanism of action, we examined viability of human fibroblast cells exposed to a Q antibiotic, ciprofloxacin (CPFX), and measured lipid peroxidation and total glutathione (GSH) levels, and activities of catalase (Cat), superoxide dismutases (SODs), glutathione peroxidase (GPx). The effects of vitamin E pretreatment on those parameters were also examined. Our results showed that the effect of CPFX on the viability of the cells, as determined by neutral red uptake assay, was time dependent. Cytotoxicity was not observed in the concentration range of 0.0129-0.387 mM CPFX when the cells were incubated for 24 hours. However, significant level of cytotoxicity was observed at concentrations 0.129 and 0.194 mM, and >0.129 mM, following 48 and 72 hours of exposure, respectively. When the cells were exposed to 0.194 mM CPFX for 48 hours, the level of lipid peroxidation increased and the content of total GSH decreased significantly; activities of total SOD, Mn SOD and CuZn SOD did not change; the decrease observed in the activity of Cat was not significant; and the activity of GPx was highly variable. Vitamin E pretreatment of the cells provided significant protection against CPFX-induced cytotoxicity; lowered the level of lipid peroxidation significantly, but increased the total GSH content only moderately; no change was observed in the activities of Cat and total SOD, but a significant increase in Mn SOD and a significant decrease in CuZn SOD were noticed. These results suggested that CPFX-induced cytotoxicity on human fibroblast cell cultures is related to oxidative stress, and vitamin E pretreatment can afford a protection. PMID- 12540035 TI - Comparative study of chronic toxic effects of daunorubicin and doxorubicin in rabbits. AB - This study compares the chronic toxicity of two anthracyclines--daunorubicin and doxorubicin, commonly used for induction of anthracycline cardiomyopathy in the rabbit model. Such a comparative study has not been published until now. Both drugs were administered intravenously to male Chinchilla rabbits in doses at 3 mg/kg (50 mg/m2) once weekly for 10 weeks. Selected biochemical, haematological and cardiovascular parameters and body weights were regularly monitored; additionally, a histological evaluation of heart, kidney and liver was performed at the end of the experiment. In the daunorubicin group, there were marked signs of the progressive development of heart failure, like the significant increases of the pre-ejection period/left ventricular ejection time index values (up to 134%)--and histological changes within the myocardium were also observed. On the other hand, the 10-week doxorubicin administration did not cause these changes that are typical for heart injury. Haematotoxicity, manifested particularly by aplastic anaemia, was apparent in both the experimental groups. Significant body weight loss (by 45.2%) and high premature mortality (100% versus 36.4%) reflected a greater general toxicity, especially nephrotoxicity of doxorubicin in comparison with daunorubicin. Further studies are necessary to find a possible explanation for these findings. PMID- 12540036 TI - Modulation of antioxidant defence system by dietary fat in rats intoxicated with o-toluidine. AB - o-Toluidine was administered to rats in the diet for four weeks at levels approximately 40, 80 and 160 mg/kg b.w. per day. Two types of diet have been used, standard (4% fat) and high fat (14% fat). Activity of antioxidant enzymes, level of glutathione and thiobarbituric acid reactive substances were measured in liver. Glutathione peroxidase was significantly increased in all treated groups while glutathione S-transferase and glutathione reductase were elevated in rats fed high-fat diet. o-Toluidine slightly enhanced catalase activity regardless of the kind of diet. Superoxide dismutase was the only enzyme whose activity was lowered in almost all treated groups. Enzymatic and nonenzymatic microsomal lipid peroxidation was enhanced 2- to 3-fold in both diet groups. Reduced glutathione level in liver was 2.3- to 4.0-fold increased in all treated groups. Our findings indicate that free radical processes can be involved in the toxic effects of o toluidine and dietary fat can modify the response of some antioxidant enzymes to this compound. PMID- 12540037 TI - Pulmonary toxicity of dust generated during weaving of carpets. AB - The dust generated during weaving (carpet dust) tibbati, knotted and tuffted carpets in carpet industry was studied for its toxicity in vitro and in vivo. Carpet dust (0.5, 1.0, 2.5 and 5.0 mg/1 x 10(6) cells) caused in vitro cytotoxicity in rat alveolar macrophages (AM) in a concentration-dependent manner. The cytotoxic, inflammatory and oxidative responses were observed in bronchoalveolar lavage fluid (BALF) of rats at 1, 4, 8 and 16 days after exposure. Rats were intratracheally exposed at 5 mg/rat individually to all three types of carpet dust. All types of carpet dusts produced increased AM, lymphocytes (PMN) population in BALF suggesting their inflammatory reactions. Cytotoxic nature of carpet dust was shown by the increased activities of lactate dehydrogenase (LDH) and acid phosphatase (AP) in BALF. Increased AM population and in vitro cytotoxicity due to carpet dusts have shown some correlation with the levels of LDH and AP activities in BALF. The gradual enhanced profile of hydrogen peroxide (H2O2) and nitric oxide (NO) along with depletion of reduced glutathione (GSH) in AM due to these carpet dusts are suggestive of their oxidant nature. The enzyme activities of both glutathione peroxidase (GPx) and glutathione reductase (GR) in AM were marginally reduced in exposed rats. In conclusion, the data suggest the cytotoxic, inflammatory and oxidant nature of carpet dusts. It is extrapolated that health effects on carpet weavers would be associated with the concentration and nature of airborne dust generated during weaving of carpets. PMID- 12540038 TI - Effects of ascorbic acid and alpha-tocopherol on arsenic-induced oxidative stress. AB - Arsenic is an ubiquitous element in the environment causing oxidative burst in the exposed individuals leading to tissue damage. Antioxidants have long been known to reduce the free radical-mediated oxidative stress. Therefore, the present study was designed to determine whether supplementation of alpha tocopherol (400 mg/kg body weight) and ascorbic acid (200 mg/kg body weight) to arsenic-intoxicated rats (100 ppm in drinking water) for 30 days affords protection against the oxidative stress caused by the metalloid. The arsenic treated rats showed elevated levels of lipid peroxide, decreased levels of non enzymatic antioxidants and activities of enzymatic antioxidants. Administration of alpha-tocopherol and ascorbic acid to arsenic-exposed rats showed a decrease in the level of lipid peroxidation (LPO) and enhanced levels of total sulfhydryls, reduced glutathione, ascorbic acid and alpha-tocopherol and so do the activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glucose-6-phosphate dehydrogenase to near normal. These findings suggest that alpha-tocopherol and ascorbic acid prevent LPO and protect the antioxidant system in arsenic-intoxicated rats. PMID- 12540039 TI - The effects of organophosphate insecticide methidathion on lipid peroxidation and anti-oxidant enzymes in rat erythrocytes: role of vitamins E and C. AB - The effects of organophosphate insecticide methidathion (MD) on lipid peroxidation and anti-oxidant enzymes and the ameliorating effects of a combination of vitamins E and C against MD toxicity were evaluated in rat erythrocytes. Experimental groups were: control group, MD-treated group (MD), and MD + vitamin E + vitamin C-treated group (MD + Vit). MD and MD + Vit groups were treated orally with a single dose of 8 mg/kg MD body weight at 0 hour. Vitamins E and C were injected at doses of 150 mg/kg body weight, i.m. and 200 mg/kg body weight, i.p., respectively, 30 min after the treatment of MD in the MD + Vit group. Blood samples were taken 24 hours after the MD administration. The level of malondialdehyde (MDA), and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) were studied in the erythrocytes. MDA level increased significantly in the MD group compared to the control group (P < 0.05) and decreased significantly in the MD + Vit group compared to the MD group (P < 0.05). The activities of SOD, GSH-Px, and CAT decreased in the MD group compared to the control group (P < 0.05). Only GSH-Px activity increased in the MD + Vit group compared with the MD group. These results suggest that treating rats with MD increases LPO and decreases anti oxidant enzyme activities in erythrocytes. Furthermore, single-dose treatment with a combination of vitamins E and C 30 min after the administration of MD can reduce LPO caused by MD. PMID- 12540040 TI - Chemical necrotizing fasciitis due to household insecticide injection: is immediate radical surgical debridement necessary? AB - We describe two cases of chemical necrotizing fasciitis in the upper extremities, anterior chest wall and epigastric region of the abdominal wall caused by household insecticide injection. We suggest that surgical debridement can be successfully performed in the subacute period under close observation in hemodynamically stable patients. PMID- 12540041 TI - Trends and variability of ground-level O3 in Connecticut over the period 1981 1997. AB - The temporal and spatial characteristics of ground-level (tropospheric) O3 measured at 10 monitoring stations in Connecticut were studied from 1981 to 1997. To detect the O3 trend caused by changes in precursor emissions, moving average filters and a linear least-squared regression model were used to eliminate the short-term variation and effects of temperature from raw time-series O3 data. The results showed a significant decrease in the number of days exceeding the National Ambient Air Quality Standards (NAAQS) and a small change in total O3 concentration. The analysis indicated that the variation of daily maximum O3, caused by changes in emissions, explained more than 10% of the total O3 variation in Bridgeport and East Hartford during the past 17 years. Meanwhile, a strong weekly cycle was also found in O3 time-series data, resulting in lower O3 concentration in urban areas than in nonurban areas, implying that land use and land cover have impacts on the spatial distribution of ground-level O3 in Connecticut. PMID- 12540042 TI - An evaluation of the tapered element oscillating microbalance method for measuring diesel particulate emissions. AB - We determined the usefulness of tapered element oscillating microbalances (TEOMs) for researchers and engineers involved with measuring diesel particulate mass. Two different test facilities were used for generating diesel particulates and comparing the TEOM to the commonly used U.S. Environmental Protection Agency (EPA) manual filter method. The EPA method is very labor-intensive and requires long periods of time to complete. The TEOM is an attractive approach because it has the potential to reduce the amount of time and labor required in diesel testing, as well as to provide real-time particulate-mass data that are not obtainable with the EPA method. It was found that the TEOM was a precise and easy to-operate instrument that could measure the mass concentration (MC) of diesel particulate emissions in real time. Although the TEOM diesel particulate MC measurements were highly correlated with the manual filter measurements, the two techniques were not equivalent because the TEOM consistently reported MC results that were 20-25% lower than those obtained using the manual filter technique. In conclusion, the TEOM can be used to increase test-cell throughput and to measure transient values of diesel particulate emissions at sites performing diesel engine testing. However, unless EPA is able to certify the TEOM as an equivalent method, it cannot replace the manual filter method for diesel certification work. PMID- 12540043 TI - Removal of PCDD/F from incinerator flue gases by entrained-phase adsorption. AB - The emission abatement of polychlorinated dioxins and furans (PCDD/F) Issued from municipal solid waste incineration is growing in importance because of more stringent emission standards and general concern about their toxic characteristics. These substances cannot be separated by conventional gas cleanup processes but are successfully removed through adsorption onto carbonaceous materials. The simplest technique is the entrained-phase injection of pulverized adsorbents in the flue gas, followed by fabric filter separation. The various related techniques are briefly reviewed here. Operating conditions and results obtained from Flemish MSWIs are given. The results illustrate the excellent overall removal efficiency. Furans are adsorbed to a slightly higher extent than dioxins. PCDD/F removal by carbonaceous adsorbents is thereafter modeled from first principles for the contribution of both entrained-phase (eta1) and cake filtration (eta2) to the overall efficiency (etaT). Application of the model equations and comparison of measured and predicted overall efficiencies for the Flemish municipal solid waste incinerators (MSWIs) demonstrate that the approach is meaningful and that the dominant parameters are the operating temperature, the dosage and activity of adsorbent, and the fraction of adsorbent in the filter cake. The model equations enable the MSWI operators to predict the adsorption efficiencies for any combination of operating parameters and to assess the sensitivity of the process to varying operating conditions. PMID- 12540044 TI - Long-term operation of a biofilter for simultaneous removal of H2S and NH3. AB - Simultaneous removal of NH3 and H2S was investigated using two types of biofilters--one packed with wood chips and the other with granular activated carbon (GAC). Experimental tests and measurements included analyses of removal efficiency (RE), metabolic products, and results of long-term operation (around 240 days). The REs for NH3 and H2S were 92 and 99.9%, respectively, before deactivation. After deactivation, the RE for NH3 and H2S were decreased to 30-50% and 75%, respectively. The activity of nitrifying bacteria was inhibited by high concentrations of H2S (over 200 ppm) but recovered gradually after H2S addition was ceased. However, the Thiobacillus thioparus as sulfur oxidizing bacteria did not show inhibition at the NH3 concentration under 150-ppm conditions. The deactivation of the biofilter was caused by metabolic products [elemental sulfur and (NH4)2SO4] accumulating on the packing materials during the extended operation. The removal capacities for NH3 and H2S were 6.0-8.0 and 45-75 mg N, S/L/hr, respectively. PMID- 12540045 TI - Controlling air pollution from passenger ferries: cost-effectiveness of seven technological options. AB - Continued interest in improving air quality in the United States along with renewed interest in the expansion of urban passenger ferry service has created concern about air pollution from ferry vessels. This paper presents a methodology for estimating the air pollution emissions from passenger ferries and the costs of emissions control strategies. The methodology is used to estimate the emissions and costs of retrofitting or re-powering ferries with seven technological options (combinations of propulsion and emission control systems) onto three vessels currently in service in San Francisco Bay. The technologies include improved engine design, cleaner fuels (including natural gas), and exhaust gas cleanup devices. The three vessels span a range of ages and technologies, from a 25-year-old monohull to a modern, high-speed catamaran built only four years ago. By looking at a range of technologies, vessel designs, and service conditions, a sense of the broader implications of controlling emissions from passenger ferries across a range of vessels and service profiles is provided. Tier 2-certified engines are the most cost-effective choice, but all options are cost-effective relative to other emission control strategies already in place in the transportation system. PMID- 12540046 TI - Determination of atmospheric nitrogen input to Lake Greenwood, South Carolina--I. PM measurements. AB - The Reedy River branch of Lake Greenwood, SC, has repeatedly experienced summertime algal blooms, upsetting the natural system. This lake's shallow depth makes It susceptible to atmospheric influence. A series of experiments were carried out in order to investigate the effect of atmospheric nitrogen deposition into the lake. Nitrogen was examined because of the insignificant phosphorus dry atmospheric flux and the unique nutrient demands of the dominant algae (Pithophora oedogonia) contributing to the blooms. In this paper, results are presented, of the experimental measurement of coarse and fine atmospheric concentrations of atmospheric particulate nitrogen adjacent to and in the watershed of the Reedy River (downtown Greenville) and Lake Greenwood. Experiments were carried out during four 24-hr periods in January 2001 and again during four 24-hr periods in March 2001. Results are presented here for atmospheric particulate nitrogen as well as other constituents of the airborne aerosol. Mass concentrations of PM2.5 averaged 14.0 and 21 microg/m3 for Lake Greenwood and downtown Greenville, respectively. Mass concentrations of total suspended particulates (TSP) averaged 22.6 and 38.5 microg/m3 for Lake Greenwood and downtown Greenville, respectively. This ambient aerosol concentration was apportioned to its chemical constituents, and the greatest contributors to PM2.5 mass were organics (45 and 42% for downtown Greenville and the lake, respectively) and sulfate (14.1 and 19.7% for downtown Greenville and the lake, respectively). The information gathered here, despite its episodic nature, is important not only in determining atmospheric nitrogen but also in documenting the composition of aerosol in South Carolina, which so far has not been studied. In a companion paper, results for gaseous pollutants as well as thermodynamic modeling of the aerosol and nitrogen flux determinations are presented. PMID- 12540047 TI - Rapidly locating and characterizing pollutant releases in buildings. AB - Releases of airborne contaminants in or near a building can lead to significant human exposures unless prompt response measures are taken. However, possible responses can include conflicting strategies, such as shutting the ventilation system off versus running it in a purge mode or having occupants evacuate versus sheltering in place. The proper choice depends in part on knowing the source locations, the amounts released, and the likely future dispersion routes of the pollutants. We present an approach that estimates this information in real time. It applies Bayesian statistics to interpret measurements of airborne pollutant concentrations from multiple sensors placed in the building and computes best estimates and uncertainties of the release conditions. The algorithm is fast, capable of continuously updating the estimates as measurements stream in from sensors. We demonstrate the approach using a hypothetical pollutant release in a five-room building. Unknowns to the interpretation algorithm include location, duration, and strength of the source, and some building and weather conditions. Two sensor sampling plans and three levels of data quality are examined. Data interpretation in all examples is rapid; however, locating and characterizing the source with high probability depends on the amount and quality of data and the sampling plan. PMID- 12540048 TI - Methodology for siting ambient air monitors at the neighborhood scale. AB - In siting a monitor to measure compliance with U.S. National Ambient Air Quality Standards (NAAQS) for particulate matter (PM), there is a need to characterize variations in PM concentration within a neighborhood-scale region to achieve monitor siting objectives. A simple methodology is provided here for the selection of a neighborhood-scale site for meeting either of the two objectives identified for PM monitoring. This methodology is based on analyzing middle-scale (from 100 to 500 m) data from within the area of interest. The required data can be obtained from widely available dispersion models and emissions databases. The performance of the siting methodology was evaluated in a neighborhood-scale field study conducted in Hudson County, NJ, to characterize the area's inhalable particulate (PM10) concentrations. Air monitors were located within a 2- by 2-km area in the vicinity of the Lincoln Tunnel entrance in Hudson County. Results indicate the siting methodology performed well, providing a positive relationship between the predicted concentration rank at each site and the actual rank experienced during the field study. Also discussed are factors that adversely affected the predictive capabilities of the model. PMID- 12540049 TI - Tau and Alzheimer's disease: the long road to anti-tangle therapeutics. PMID- 12540050 TI - Neurofibrillary degeneration can be arrested in an in vivo cellular model of human tauopathy by application of a compound which inhibits tau filament formation in vitro. AB - Although tau filament formation is a central event in familial tauopathies and Alzheimer's disease (AD), the cellular consequences of neurofibrillary tangle (NFT) formation are poorly understood because of the unavailability of mammalian in vivo cellular models of neurofibrillary degeneration (NFD). We have shown that human tau forms filaments and is associated with cytodegeneration when overexpressed chronically in identified neurons (ABCs) in the lamprey central nervous system (CNS). In this model, degeneration occurs according to a stereotyped sequence that closely resembles the pattern seen in tangle-bearing neurons in AD, with both tau deposition and fragmentation beginning in distal dendrites and progressing proximally over time. This sequence has been divided into four stages ranging from (1) mild beading of terminal dendrites only through (4) extensive dendritic fragmentation and loss. Here, we show that lipid-soluble, low-molecular-weight (approx 300 Da) proprietary compounds that have been demonstrated to block tau filament formation in vitro can significantly retard the progressive degeneration of ABCs that express human tau23. Bath application of one of these compounds for periods of up to 50 d after plasmid injection prevented degeneration beyond stage 2 in 90% of all treated cells, whereas over half of control cells showed severe degeneration by this time. This provides the first in vivo experimental evidence directly supporting a causal role for tau filament formation in the pathogenesis of NFD and suggests that intensive effort toward developing therapeutic agents for AD and other NFDs targeted at blocking tau filament formation is warranted. PMID- 12540051 TI - Discovery of compounds that will prevent tau pathology. AB - Tau is certainly a reasonable target for the development of compounds to prevent neurofibrillary pathology, particularly in the fronto-temporal dementias. Although the mechanism of the filamentous accumulations remains unclear, sufficient knowledge is in place to move forward with high throughput screens. In fact, the development of compounds from such screens will ultimately be the only way to validate the target. The dichotomy for such screens is that in vitro screens are easier to design, but require more assumptions as to the mechanism, in contrast to cell-based screens that are more difficult to design, but make fewer assumptions about mechanism. We have designed a moderate throughput for tau binding that relies on fluorescence detection in living cells and an in vitro cdk5/p25 tau phosphorylation high throughput screen. PMID- 12540052 TI - Cdk5 as a drug target for the treatment of Alzheimer's disease. AB - Cyclin-dependent kinase-5 (cdk5) is suggested to play a role in tau phosphorylation and contribute to the pathogenesis of Alzheimer's disease (AD). One of its activators, p25, is dramatically increased in AD brains where p25 and cdk5 are colocalized with neurofibrillary tangles. Several animal models have shown a correlation of p25/cdk5 activities with tau phosphorylation. Overexpression of p25/cdk5 in nueronal cultures not only leads to tau phosphorylation but also cytoskeletal abnormalities and neurodegeneration. Therefore, cdk5 kinase inhibitors are potential therapeutic agents for the treatment of AD. Availability of potent, selective, brain permeable cdk5 inhibitors and relevant animal models in which their efficacy can be treated will be critical in the development of these inhibitors. PMID- 12540053 TI - Pin1: a therapeutic target in Alzheimer neurodegeneration. AB - In Alzheimer's disease, the peptidyl prolyl cis/trans isomerase Pin1 binds to phospho-Thr231 on Tau proteins and, hence, is found within degenerating neurons, where it is associated to the large amounts of abnormally phosphorylated Tau proteins. Conversely, Pin1 may restore the tubulin polymerization function of these hyperphosphorylated Tau. In the present work, we investigated, both at the cellular and molecular levels, the role of Pin1 in Alzheimer's disease through the study of its interactions with phosphorylated Tau proteins. We also showed that in neuronal cells, Pin1 upregulates the expression of cyclin D1. This, in turn, could facilitate the transition from quiescence to the G1 phase (re-entry in cell cycle) in a neuron and, subsequently, neuronal dedifferentiation and apoptosis. The involvement of Pin1 in the G0/G1 transition in neurons points to its function as a good target for the development of new therapeutic strategies in neurodegenerative disorders. PMID- 12540054 TI - Tau neurofibrillary pathology and microtubule stability. AB - We previously reported that nonomolar concentrations of Taxol and several structurally diverse microtubule (MT)-stabilizing agents significantly enhanced the survival of neurons in the presence of fibrils of amyloid beta peptide (Abeta). Pretreatment of neurons with MT-stabilizing drugs also blocked Abeta induced activation of tau hyperphosphorylation. Although tau is a substrate for several kinases, we initially focused on cdk5, as this tau kinase has been shown to be activated in Abeta-treated neurons and Alzheimer's disease (AD) brain. In an in vitro kinase assay, Taxol inhibited activation of cdk5 by Abeta. In addition, the proposed cellular cascade in which calpain activation leads to cleavage of the cdk5 regulator, p35, to the strong kinase activator p25 was also prevented. Taxol did not directly inhibit the activity of either cdk5 or calpain, indicating that other cellular components are required for the effect of the drug on Abeta activation of tau phosphorylation. Our results suggest that drugs that interact with MTs can alter signaling events in neurons, possibly because some MTs play a role in organizing protein complexes involved in responses to Abeta. Thus the cytoskeletal network may serve as a biosensor of cellular well-being. PMID- 12540055 TI - p38 mitogen activated protein kinase as a therapeutic target for Alzheimer's disease. AB - The characteristic pathological hallmarks of Alzheimer's disease (AD) include neuritic plaques, neurofibrillary tangles, and inflammatory changes. Current therapies, such as molecules that target enhancing cholinergic activity, can improve cognitive function in the short term but, unfortunately, have no impact on progression of the disease. Although many molecular targets have been suggested to play a causative role in AD progression, clinical data demonstrating a link between the blockade of such targets and amelioration or halting of disease progression are lacking. Even so, there are many interesting candidate targets, and current research efforts in these areas promises to deliver a wealth of new possibilities for treating AD in the future. This brief review will focus on p38 mitogen-activated protein kinase as a possible target for therapeutic intervention in AD. PMID- 12540056 TI - Neuroprotective properties of valproate: potential benefit for AD and tauopathies. AB - Neuropsychiatric disturbances are extremely common in Alzheimer's disease (AD), and represent integral features of the illness, as well as appropriate targets for therapy. We are interested in designing trials aimed at preventing or delaying the emergence of psychopathology in AD. For symptomatic treatment of agitation, mood stabilizers, particularly sodium valproate, have proved to be beneficial in some patients. Since these effects take several weeks to emerge, we considered that they might be dependent on potentially neuroprotective actions of valproate, such as inhibition of apoptosis and slowing of neurofibrillary tangle formation. In this article we present the rationale for testing the neuroprotective potential of valproate experimentally in mouse models of tauopathy and in a clinical trial of patients with AD who lack psychopathology at baseline. Together, these studies will provide important tests of the hypothesis that valproate, either through inhibition of tau phosphorylation or some other mechanism, is a useful therapeutic agent to modify disease progression in AD. PMID- 12540058 TI - Organotypic slice cultures from transgenic mice as disease model systems. AB - Organotypic slice cultures have been prepared from the brains of transgenic mice with Alzheimer's disease-type pathology. Cell types within the slice undergo differentiation and slices can be maintained in culture for up to 6 mo when prepared from young neonates. Slices have been prepared from mice overexpressing genes of relevance to Alzheimer's disease, including mutant or wild-type tau. Neurons in these slices develop neurons that are immunoreactive for a number of markers of abnormal tau. Organotypic slice models are currently being used to test the impact of tangle enhancers or inhibitors as a prescreen for efficacy before testing drugs in vivo. PMID- 12540057 TI - Cellular models for tau filament assembly. AB - Filamentous inclusions made of phosphorylated tau constitute a neuropathologic hallmark of Alzheimer's disease and related disorders. Because abnormal accumulation of tau correlates with the decline in cognitive function, it is conceivable that effective intervention at the early stage of tau filament assembly could impede the progression of these neurodegnerative diseases. Cellular models recapitulating the tau aberration are useful for screening and identifying compounds that are capable of interfering with tau aggregation in a cost-effective manner. To develop such cell culture models, we have established from human neuroglioma [H4] and neuronal [BE(2)-M17D] cells conditional transfectants whose transgenic expression of wild-type or mutant tau via inducible expression mechanisms leads to tau aggregation and filament assembly. PMID- 12540059 TI - In vivo brain imaging of tangle burden in humans. AB - Cerebral neurofibrillary tangles (NFTs) accumulate in a predictable sequence decades before the clinical symptoms of Alzheimer's disease emerge, and the degree of tangle degeneration correlates with the severity of cognitive impairment. A valid in vivo marker of tangle burden, therefore, would be useful for presymptomatic and symptomatic disease detection and treatment monitoring. Recent advances using positron emission tomography (PET) indicate the feasibility of in vivo imaging that provides a combined signal of both neurofibrillary tangles and senile plaques. Such results are encouraging that a tangle-specific marker will be found; however, several methodological issues first need to be addressed, including scanner spatial resolution in the relatively small brain regions where tangles accumulate. NFT-specific imaging probes will need to be lipophilic in order to cross the blood-brain barrier and neuronal membranes and have a high binding affinity to NFTs with minimal nonspecific binding, which would result in a high signal-to-background ratio in PET images. PMID- 12540060 TI - Tau therapeutics for Alzheimer's disease: the promise and the challenges. AB - The pathological diagnosis of Alzheimer's disease (AD) depends on the presence of plaques consisting of the beta-amyloid peptide as well as neurofibrillary tangles consisting of paired helical filaments (PHFs) of the tau (tau) protein. The role of each type of pathology in the pathogenesis and progression of AD remains unclear. Previous hypotheses suggested that these two processes were independent, whereas more recent data suggest that there may be a bidirectional interaction between these two pathological processes. The identification of the neurotoxic effects of beta-amyloid and the discovery of mutations responsible for early onset Alzheimer's disease (EOAD) and their linkage to beta-amyloid overproduction, has made the amyloid hypothesis of AD the predominant influence for therapeutic targets. Several approaches have emerged from preclinical testing and have entered early phases of clinical developments. The recent identification of tau mutations and their linkage to progressive neurodegenerative disorders provides a counterbalancing influence on the search for therapeutic targets for AD. Therapeutic approaches that are targeted to either beta-amyloid or tau share certain features at the level of pharmacology and will face many of the same challenges as they progress through drug development paradigms. The aim of this article is to provide a brief overview of some of the commonalities and the challenges faced by tau-related therapeutic strategies. The issues discussed in this article are not exhaustively dealt with in either scope or detail. PMID- 12540061 TI - Tau as a drug target in Alzheimer's disease. PMID- 12540062 TI - World report on violence and health: engaging utopia through evidence. PMID- 12540063 TI - Treatment of chronic hepatitis B in india: what is the cost of life? PMID- 12540064 TI - Treatment of chronic hepatitis B with interferon-alpha: cost-effectiveness in developing countries. AB - BACKGROUND: Treatment with interferon-alpha (IFN) has been shown to be cost effective in developed countries. However, cost-effectiveness In developing countries such as India has not been studied. METHODS: Using the Markov transitional probability model, we studied two cohorts of young patients (30 years of age) with chronic hepatitis B, one untreated and the other treated with interferon (IFN), 5 million units daily for 16 weeks, with evidence of viral replication and chronic hepatitis, but not cirrhosis, and were followed up over a 30-year period. Rates of disease progression, efficacy of IFN and quality of life associated with various disease states were estimated from the available literature. Direct costs were estimated using Indian prices of IFN and from the usual costs of medical treatment in India based on expert opinion. Unrelated mortality rates were modelled on age-specific death rates of the general population. The efficacy of IFN was judged In terms of extra life-years and quality-adjusted life-years (QALY) gained, and marginal cost-effectiveness and cost-utility. Several sensitivity analyses, both undiscounted and with discounted analyses, were done. RESULTS: At the end of the 30-year period, fewer patients in the IFN-treated group developed cirrhosis or decompensated cirrhosis, or were dead. The average life span of the treated cohort was 25.14 years, a gain of 0.6 years over the untreated cohort (24.54years). The QALY lived bythetwocohortswere 23.69 and 22.75 years, respectively, representing a gain of 0.94 years for the IFN-treated group. The cost Incurred by the IFN-treated group was Rs 300,000, and that for the untreated cohort was Rs 40 700, a substantial difference. Using the baseline estimates, undiscounted costs per year of life gained and per QALY gained were Rs 432,500 and Rs 276,900, respectively; these estimates are 20.5 and 13.1 times the per capita gross national income of the Indian population. Sensitivity analyses showed that changes in various parameters led to only minor changes in these estimates. Use of discounting led to an increase in marginal cost per life-year or QALY gained. CONCLUSION: In developing countries with a low per capita Income, IFN therapy for chronic hepatitis B may not be cost-effective. A careful consideration of cost-effectiveness is therefore essential before Instituting IFN therapy in patients with chronic hepatitis B In such populations. PMID- 12540065 TI - The clinical spectrum of fibrocalculous pancreatic diabetes in north India. AB - BACKGROUND: Fibrocalculous pancreatic diabetes (FCPD) is a secondary form of diabetes, unique to tropical countries. In earlier reports, patients with FCPD had severe insulin-requiring diabetes, malnutrition and a dismal prognosis. With Improvements in nutrition and medical care, the presentation and prognosis of FCPD may have changed. We report on the clinical profile and prognosis of a cohort of FCPD patients from north India and compare our findings with earlier reports. METHODS: Eighty consecutive FCPD patients who presented to the Diabetes, Gastroenterology and Surgical Gastroenterology services were evaluated for their nutritional status, clinical presentation, beta-cell function (fasting C-peptide) and exocrine function (faecal chymotrypsin). All patients diagnosed between 1994 and 2000 (n = 32) were followed prospectively for weight gain and glycaemic control. RESULTS: Only 55% of FCPD patients had a low body mass index (< 18 kg/m2). At the time of diagnosis of diabetes, only 26 (33%) patients presented with severe insulin-requiring diabetes; these patients were younger [23.7 (8.3) years v. 28.7 (10.6) years, p = 0.04], and had higher haemoglobin A1c [9.7 (3.8)% v. 7.3 (2.6)%, p = 0.005] than those requiring diet control or oral hypoglycaemic agents. FCPD patients had a wide range of fasting serum C-peptide (0.03-0.76 nmol/L). C-peptide was negatively associated with increasing duration of diabetes (r = -0.48, p = 0.001), but there was no correlation with faecal chymotrypsin. On prospective follow up (mean 2.3 years), there was significant improvement in body mass index [19.4 (2.9) kg/m2 v. 17.0 (3.7) kg/m2, p < 0.01] and haemoglobin A,c [6.4 (1.6)% v. 8.0 (3.0)%, p < 0.001]. CONCLUSION: FCPD patients differed from those described in earlier reports in many respects, Including improved nutritional status, a wide range of 3cell function and a more favourable prognosis. PMID- 12540066 TI - Explanatory models and common mental disorders among patients with unexplained somatic symptoms attending a primary care facility in Tamil Nadu. AB - BACKGROUND: Patients with unexplained somatic symptoms are commonly seen in primary care. We assessed the explanatory models and common mental disorders in patients with unexplained somatic symptoms attending a primary care facility in a rural area of south India. METHODS: One hundred consecutive patients diagnosed to have unexplained somatic symptoms attending a primary care facility were examined. The Tamil version of the Revised Clinical Interview Schedule was used to assess common mental disorders and the Tamil version of the Short Explanatory Model Interview was used to assess their explanatory models. RESULTS: Ninety eight patients thought that their problem was serious. Sixty-nine, 41 and 40 claimed that it affected their work, family and social lives, respectively. Forty two of the patients believed in black magic. Forty-four patients satisfied the ICD-10 PC criteria for common mental disorders. Depression was the commonest diagnosis (63.6%). Patients with psychiatric diagnoses admitted to emotional problems (chi2 = 46.9; p < 0.001), and felt that their problems affected their family (chi2 = 38.5; p < 0.001), social life (chi2 = 28.6; p < 0.001) and work (chi2 = 34.4; p < 0.001). More patients who satisfied criteria for psychiatric diagnosis felt that their problems were moderate or very serious (chi2 = 21.5; p < 0.001) and they feared they might become disabled or die (chi2 = 12.2; p < 0.001). Psychiatric syndromes were significantly associated with belief in black magic (chi2 = 7.01; p < 0.01). These associations remained statstically significant after adjusting for age, gender, literacy and socioeconomic indices using logistic regression. CONCLUSION: The majority of patients held strong beliefs regarding the physical nature of their complaints, believed In the serious nature of the problem and feared disability or death. There is a need to elicit specific explanatory models regarding the nature of illness in patients who present with somatic symptoms without organic causes. Understanding the patient's perspectives is a prerequisite for providing the necessary treatment and to dispel fears. PMID- 12540067 TI - Acute fatty liver of pregnancy: a report of two cases. AB - Acute fatty liver of pregnancy is an uncommon, potentially fatal disorder. Between 1998 and 2000, two patients with acute fatty liver of pregnancy presented at the Christian Medical College Hospital, Vellore. Both patients were in the thirty-sixth week of pregnancy. jaundice and encephalopathy were the predominant symptoms. Both the mothers died after they delivered a stillborn Infant each. The maternal deaths were due to multiorgan failure and/or postpartum haemorrhage and sepsis. The route of delivery was vaginal in both the patients. Extrahepatic and metabolic complications in both cases Included renal failure, sepsis, hypoglycaemia, disseminated intravascular coagulation and gastrointestinal bleeding. Liver biopsy done in both patients was consistent with the diagnosis of acute fatty liver of pregnancy. To the best of our knowledge, this is the first report from India on acute fatty liver of pregnancy. PMID- 12540068 TI - Human prion diseases. AB - Prion diseases is another name for a group of 'transmissible spongiform encephalopathies'. Creutzfeldt-Jakob disease, the first prion disease described in humans, occurs in sporadic, familial or iatrogenic form. Other transmissible spongiform encephalopathies in humans such as familial Creutzfeldt-]akob disease, Gerstmann-Straussler-Scheinker disease and fatal familial Insomnia have been shown to be associated with specific prion protein gene mutations. In 1996, a new variant of Creutzfeldt-Jakob disease was reported in the United Kingdom among young patients with unusual clinical features and unique neuropathological findings. This new form could be due to transmission to humans of the agent causing bovine spongiform encephalopathy. While examination of brain tissue is the key to making a diagnosis, it is not always possible antemortem. Immunological tests such as ELISA or western blot assays along with tests for 1 4 3-3 protein in the cerebrospinal fluid remain the main tools of diagnosis. Conventional disinfection and sterilization practices are Ineffective for these agents. The unusual properties of prions pose a challenge for treatment, surveillance and control of these diseases. PMID- 12540069 TI - Hormone replacement therapy: need for reappraisal. PMID- 12540070 TI - Preventing infection during manual removal of the placenta. PMID- 12540071 TI - Criminal medical negligence: the need for a re-look. PMID- 12540072 TI - Suicide: rates and methods before, during and after the war in Croatia (1985 2000). PMID- 12540073 TI - Medical genetics education in India. PMID- 12540074 TI - Telepathology at Tata Memorial Hospital, Mumbai and Barshi, a rural centre in Maharashtra. PMID- 12540075 TI - "All you need is love". PMID- 12540076 TI - Perioperative complications of total ankle arthroplasty. AB - A retrospective radiographic and chart review was performed for the initial 50 patients who underwent Agility (DePuy, Warsaw IN) total ankle arthroplasty by the senior author (M.S.M.). The review focused on the perioperative complications of nerve or tendon lacerations, intraoperative fractures, acute deep infections, wound complications and component positioning. Major wound complications were defined as those requiring a soft-tissue coverage procedure. Minor wound complications did not require soft tissue coverage and included wound breakdowns, wound edge necrosis, and superficial infections. The immediate mortise and lateral postoperative radiographs were reviewed to measure component positioning. The patients were divided into two groups to compare the initial 25 patients (Group A) with the subsequent 25 patients (Group B). There were no major wound complications in either group. Minor wound complications decreased from six in Group A to two in Group B. There were four lacerations (flexor hallucis longus, posterior tibial tendon, deep peroneal nerve, and superficial peroneal nerve), all occurring in Group A. Five patients sustained intraoperative fractures in Group A, as compared with two fractures in Group B. The number of components varying greater than 4 degrees from neutral as measured by the lateral talar, lateral tibial and mortise tibial component angles decreased by 9% from Group A to Group B. The only tibial component to be placed in more than 4 degrees of valgus occurred in Group A. It seems that a notable learning curve exists in the performance of total ankle arthroplasty as demonstrated by a comparison of the initial 25 patients with the subsequent 25 patients performed by one orthopaedic surgeon. This improvement most likely resulted from the use of enhanced techniques and further training with the prosthesis. This information can be used as a teaching tool to decrease the incidence of complications for surgeons performing their initial arthroplasties with this potentially technically demanding procedure. PMID- 12540077 TI - Correction of ankle and hindfoot deformities by supramalleolar osteotomy. AB - Ankle and hindfoot deformities in 11 patients with a mean age of 15 (range, four to 35) have been gradually corrected by an llizarov external fixator. In all patients, supramalleolar osteotomy was utilized by percutaneous drilling and osteotomy technique. The patients were evaluated clinically by the AOFAS scoring system and radiologically by malorientation and malalignment tests described by Paley. The purpose of the current study is to evaluate the efficacy of supramalleolar osteotomy and correction by an Ilizarov device in treating hindfoot and ankle deformities. The mean external fixation period was five (2.5 to eight) months. The mean interval between the operation and last follow-up examination was 19 (13 to 26) months. The patients improved an average of 30 points after the operation regarding to AOFAS score. A plantigrade foot was achieved in the whole group. Radiologically, no malalignment or malorientation was measured. Equal leg lengths were achieved in all nine patients with one exception. Complications were graded after Paley as minor problems in 11 patients and as obstacles requiring surgical interventions in four patients. Sequelae remained in only one patient. Ankle and hindfoot deformities caused by various etiologies are complicated by poor soft tissue condition and a short foot. A supramalleolar osteotomy and Ilizarov external fixator can safely and effectively correct these deformities. PMID- 12540078 TI - Scarf osteotomy for hallux valgus repair: the dark side. AB - HYPOTHESIS/PURPOSE: The Scarf osteotomy has gained popularity as treatment of choice in parts of Europe and is based on sound structural principles. The excellent results reported by others could not, however, be reproduced by the author and the results are presented. METHODS: From January 1997 to June 1997 the Scarf osteotomy was selected in 20 consecutive patients (12 female and eight male patients, ages 18 to 60, mean: 41 years) with moderate metatarsus primus varus (IMA 13 to 20 degrees) and hallux valgus deformities (less than 40 degrees). The AOFAS Hallux Metatarsophalangeal-Interphalangeal Scale, visual analog scale and patient satisfaction were monitored prior to surgery, six and 12 months post-op. The patients were treated in a short leg cast, non-WB for two weeks followed by four weeks partial WB in a cast shoe. Routine post-bunion rehabilitation followed once the radiological and clinical diagnosis of healing was made. RESULTS: Multiple complications were encountered. The most common was "troughing" of the metatarsal with loss of height. This occurred in seven patients (35%). Other complications include delayed union (5%), rotational malunion (30%), proximal fracture (10%), infection (5%) and early recurrence of deformity in 25%. All 20 patients were available for follow-up at six months, and 19 of 20 at 12 months. The AOFAS score pre-op was a mean of 53. At six months a mean of 54 (19 to 69) and at 12 months 62 (24-100). Forty-five percent (9/20) were unsatisfied at one year and would not recommend the surgery to a friend. CONCLUSIONS/SIGNIFICANCE: The Scarf osteotomy has multiple potential pitfalls and should probably be reserved for moderate bunions in young people with good bone quality. There are multiple potential problems and the salvage of a failed Scarf osteotomy is difficult. PMID- 12540079 TI - The Ludloff metatarsal osteotomy: guidelines for optimal correction based on a geometric analysis conducted on a sawbone model. AB - Ideally, osteotomy for hallux valgus deformities corrects varus angle and pronation, while minimizing elevation, depression, and shortening. We used a serial linkage tracking device to evaluate five variations of the Ludloff osteotomy, a stable proximal metatarsal osteotomy that incorporates an oblique dorsal to plantar cut and a lateral swing or pivoting correction of the dorsal fragment. A neutral osteotomy (perpendicular to the sagittal plane) yielded the greatest correction (14 degrees) but with shortening (average, 2.85 mm), elevation (average, 1.36 mm), and additional pronation (average, 1.88 degrees). The same osteotomy angled 10 degrees plantarly with 8 degrees of correction produced an average of 1.57 degrees of supination, 1.22 mm of depression and, 0.54 mm of shortening. Additional plantar inclination (20 degrees) and angular correction (16 degrees) yielded increased depression, supination, and shortening. The current findings provide guidelines to achieve the desired correction and rotation and suggest that optimal results can be obtained by performing this osteotomy angled 10 degrees plantarly with a correction of 8 degrees to 16 degrees. PMID- 12540080 TI - The influence of rheumatoid chemotherapy, age, and presence of rheumatoid nodules on postoperative complications in rheumatoid foot and ankle surgery: analysis of 725 procedures in 104 patients [corrected]. AB - The records of 104 patients who underwent reconstructive foot and ankle surgery for deformities secondary to rheumatoid arthritis were reviewed. The use of rheumatoid chemotherapeutic agents, age, sex, rheumatoid nodule status, and the number of concurrent surgical procedures performed was analyzed to determine any association with the postoperative outcome for wound healing and infectious complications. The 104 patients, ranging in age from 23 to 83 years, underwent 725 operative procedures. An overall 32% complication rate was recorded. Analysis of five specific rheumatoid chemotherapeutic agents (NSAIDs, steroids, methotrexate, hydroxychloroquine, gold), age, sex, number of operative procedures performed, and presence of rheumatoid nodules, either alone or in combination, failed to prove a statistical association with either a healing or infectious postoperative complication. PMID- 12540081 TI - Plantar pressures in fiberglass total contact casts vs. a new diabetic walking boot. AB - The objective of this study was to determine in healthy individuals whether the Bledsoe Diabetic Conformer Boot reduces plantar pressures as well as a fiberglass total contact cast. Eighteen normal subjects, without any prior foot or ankle problems, were recruited for this study. Plantar pressures were measured using the Novel Pedar in-shoe pressure measurement system. The results of this study demonstrate that in individuals without foot deformities, the Bledsoe Diabetic Conformer Boot performs as well as, and in many parameters, even better than a fiberglass total contact cast with respect to reducing the forces and pressures on the plantar surface of the foot. PMID- 12540082 TI - Evaluation of the validity of the AOFAS Clinical Rating Systems by correlation to the SF-36. AB - This study evaluates the validity of the AOFAS Clinical Rating Systems by examining their level of correlation to the Medical Outcomes Study Short Form-36 (SF-36) in patients with foot and ankle complaints. The SF-36 is an extensively validated outcomes tool that has been used as a benchmark in examining the validity of outcomes instruments designed for the upper extremity, knee, shoulder, and general orthopaedic conditions. The study sample was 91 patients seen at the foot and ankle clinic of a university-based orthopaedic practice. Patients were administered both the AOFAS Clinical Rating Systems and SF-36 instruments. Pearson correlation coefficients of the AOFAS scores to the SF-36 sub-scales ranged from 0.02 to 0.36 in the overall study population. Correlation was higher for the sub-set of patients with ankle-hindfoot disorders (0.11 to 0.53) than patients with forefoot disorders (-0.05 to 0.25). The low levels of correlation seen in this study suggest poor construct validity of the AOFAS Clinical Rating Systems. PMID- 12540083 TI - The problem with measuring patient perceptions of outcome with existing outcome measures in foot and ankle surgery. AB - Quality outcome measures are the cornerstone of clinical research. A review of outcome measures used in foot and ankle surgery research reveals that the issues of validity, reliability and responsiveness of outcome measures have not been addressed. Most reports in the literature have attempted to evaluate patient perceptions of outcome following foot surgery. Underlying the many difficulties with these outcome measures is a lack of understanding of what patients perceive to be important in terms of outcome. Consequently none of the existing outcome measures can claim to be valid measures of patient perceptions of outcome, as there has been no research uncovering these perceptions. In addition, measures of general health status and quality of life in relation to outcome of foot and ankle surgery have been largely ignored to date. PMID- 12540084 TI - Anchor enhanced capsulorraphy in bunionectomies using an L-shaped capsulotomy. AB - The objective of this study was to investigate potential benefit of a suture anchor-enhanced capsulorraphy in the early maintenance of correction in bunionectomies. We compared, retrospectively, in successive series, the loss of correction of the Hallux Valgus (HV) and intermetatarsal (IM) angle, in those repaired with an L-shaped capsulorraphy enhanced with anchors to those without. Intraoperative and second week postoperative simulated weightbearing anterior posterior (AP) X-rays were used to evaluate results. By using only intraoperative and early postoperative X-rays, we should have effectively eliminated extraneous factors that might have influenced our results. A Total of 106 cases were investigated, 65 of which were repaired using anchors, the remaining 41 without. In the anchor group, 38 underwent a proximal metatarsal concentric shelf osteotomy (CSO)/modified McBride procedure, while the remaining 27 had a distal Chevron correction. In the without-anchor group, 21 had a CSO/modified McBride procedure while 20 underwent the Chevron procedure. In the without-anchor group, the average HV and IM loss of correction was 4.60 degrees (range, -2 to 21 degrees) and 0.6 degrees (range, -1 to 9 degrees) respectively. In the anchor group, the corresponding loss was 2.8 degrees (range, -3 to 17 degrees) and 0.6 degrees (range, -2 to 14 degrees) respectively. These results, when statistically analyzed, demonstrated that while the IM angle change was not statistically significant, the HV angle change was statistically significant, implying that the anchor plays a significant role in maintaining the surgical correction in both the distal Chevron and CSO/ modified McBride bunionectomies. PMID- 12540085 TI - Tendon transfer fixation in the foot and ankle: a biomechanical study evaluating two sizes of pilot holes for bioabsorbable screws. AB - The purpose of this study was to compare the initial fixation strengths of bioabsorbable screws for tendon transfers in the foot and ankle when the pilot hole size varied. A 7 x 20 mm screw was used with 5.5 mm and 6.5 mm drill holes, and a 5 x 20 mm screw was used with 3.9 mm and 4.5 mm drill holes. Biomechanical testing was performed on each tendon transfer in cadaver specimens. A paired t test showed no significant difference in pullout strength when pilot hole size varied between 79 to 93% of the screw size for the 7 mm screw and 78 to 90% of the screw size for the 5 mm screw. Previous studies have found a critical value of tendon tension equaling 50 N with passive dorsiflexion of the foot. With an average value of approximately 170 N, the 7 mm screw provided three times the requisite strength. The 5 mm screw provided 1.5 times the requisite strength, but the transfer was technically more difficult. PMID- 12540086 TI - Hallux valgus in men. Part II: First ray mobility after bunionectomy and factors associated with hallux valgus deformity. AB - PURPOSE: To determine the 1st ray mobility following a distal soft-tissue procedure with proximal osteotomy (DSTP-PMO) and any associated factors. METHODS: A retrospective study of 30 men (35 feet) was performed. First ray mobility, ankle dorsiflexion, pes planus, and metatarsus adductus were evaluated at the final follow-up. All internal fixation was routinely removed at six to eight weeks postoperatively. Standard radiographs were evaluated and angular measurements were calculated on all feet. RESULTS: The mean follow-up was 78 months. No cases of degenerative arthritis of the 1st MTC joint were noted on follow-up radiographs. DSTP-PMO resulted in a mean postoperative 1st ray mobility of 4.9 mm (range, 2.5 to 8). In those feet evaluated following bunion correction, there was no correlation with pes planus, limited ankle dorsiflexion or metatarsus adductus. The preoperative hallux valgus angle and 1-2 intermetatarsal angle correlated with toe pronation and a positive family history. Twenty-two patients had an AOFAS score of 90-100, seven of 80-89 and one less than 69. CONCLUSION: Hallux valgus in this group of male patients was not associated with limited ankle dorsiflexion or pes planus. Men with toe pronation and a positive family history had a greater hallux valgus deformity than those without after a distal soft tissue repair with proximal first metatarsal osteotomy. There was no evidence of first ray hypermobility after a DSTP-PMO. PMID- 12540087 TI - Management of large soft-tissue defects in a diabetic patient. AB - Amputation as the only option for treatment of large ulcers in a patient with diabetes is well known. This is a case presentation of a patient with large ulcers and osteomyelitis related to diabetes. He was treated with extensive debridement, segmental shortening, and wound closure. PMID- 12540088 TI - Bursitis as a cause of plantar heel pain: a case report. PMID- 12540089 TI - Technique tip: a revised method of the Cotton test for intra-operative evaluation of syndesmotic injuries. AB - The Cotton test assists the orthopaedic surgeon in the intra-operative evaluation of potential incompetence of the ankle syndesmosis. A variation of this test is proposed that simplifies this test, while providing a direct distraction force to the syndesmotic ligamentous complex. PMID- 12540090 TI - The effect of storage and lag time on MS2 bacteriophage susceptibility to ultraviolet radiation. AB - The susceptibility of MS2 bacteriophage suspensions to UV radiation was assessed using a collimated beam technique. Storage of MS2 bacteriophage cultures at 4 degrees C resulted in a decrease in phage susceptibility to UV radiation over time. After 18 days, the level of MS2 bacteriophage inactivation achieved for the range of UV doses tested decreased by 0.7 to 1.1 logs, but remained constant after that point. Changes in the protein coat of the bacteriophage, a decrease in viability over time, and an increase in coagulation may have played a role in the observed susceptibility decrease. A 2-hour lag time between the preparation of the MS2 suspension and the irradiation test also resulted in a decrease in phage susceptibility. PMID- 12540091 TI - Electroadsorption of acilan blau dye from textile effluents by using activated carbon-perlite mixtures. AB - The feasibility of the removal of dye stuffs from textile effluents by electroadsorption has been investigated. An activated carbon-perlite mixture with a ratio of 8:1 for bipolarity has been used as the adsorbent. Conventional adsorption experiments have also been conducted for comparison. A bipolar trickle reactor has been used in the electroadsorption experiments. The model wastewater has been prepared by using acilan blau dye. Initial dye concentration, bed height between the electrodes, applied potential, flowrate, and the supporting electrolyte concentration have been examined as the parameters affecting the removal efficiency. A local textile plant effluent has been treated in the optimum values of these parameters obtained from the experimental studies. Adsorption kinetics and the amount of adsorbent required to reach the maximum removal efficiency have also been investigated and mass-transfer coefficients have been calculated for adsorption and electroadsorption. The results showed that a removal efficiency of up to 100% can be achieved with energy consumption values of 1.58 kWh/m3 of wastewater treated. However, energy consumption decreases to 0.09 kWh/m3 if an exit dye concentration of 4.65 mg/L is accepted. It can be concluded from this work that this method combines all of the advantages of the activated-carbon adsorption and electrolytic methods for the removal of dyes from wastewater. PMID- 12540092 TI - Enumeration of coliform bacteria in wastewater solids using defined substrate technology. AB - Commercially available defined substrate technology for the enumeration of coliform bacteria was investigated for its applicability to waste activated sludge (WAS) solids. The defined substrate, Colilert, in conjunction with the QuantiTray enumeration system, was compared with multiple-tube fermentation (MTF). Multiple-tube fermentation analyses (U.S. Environmental Protection Agency [U.S. EPA]-accepted methods for the enumeration of coliforms in wastewater solids) were conducted on settled wastewater solids using lauryl tryptose broth as the presumptive phase and brilliant-green lactose bile broth for the confirmed phase of total coliform enumeration. Both EC medium for fecal coliform identification and EC-4-methyl-umbelliferyl-beta-D-glucuronide (MUG) media for Escherichia coli for the MTF completed phase were also conducted on the WAS. Samples analyzed included raw WAS and WAS that had been pasteurized at 70 degrees C for 15 and 30 minutes, respectively. Twenty replicates of each sample were measured to demonstrate statistical equivalence of the total coliform numbers for all three WAS samples (raw, 15-minute pasteurization, and 30-minute pasteurization). Results show that both systems provided statistically equal total coliform most probable numbers (MPNs) for all samples tested. As expected, fecal coliform numbers identified by the MTF technique were statistically higher than E. coli enumerated by Colilert-QuantiTray. However, the E. coli MPN generated by the MTF method was found to be statistically equal to the Colilert QuantiTray numbers. Because of its ease of use and reduced time to obtain the analysis, the Colilert-QuantiTray system can be useful in enumerating coliforms in WAS. However, more data are required to definitively show equivalence of the commercially available defined substrate technology to the U.S.-EPA accepted protocol. It is hoped that the results of this study will help form a database for the acceptance of the Colilert-QuantiTray system for WAS bacterial analyses. PMID- 12540093 TI - Modeling of nitrification inhibition with aniline in suspended-growth processes. AB - Nitrification inhibition due to aniline was investigated in completely mixed suspended-growth, batch, and continuous processes. Synthetic wastewater was used with aniline as the carbon source. The experiments were conducted at aniline concentrations inhibitory to nitrifier organisms. In the batch tests, degradation took place rapidly (within 4 to 6 hours) for initial aniline concentrations below 100 mg/L, with nitrification picking up as soon as the aniline concentration decreased to less than 3 to 4 mg/L. For initial aniline concentrations of 250 mg/L and higher, complete nitrification did not take place even though the aniline concentration decreased to less than 3 to 4 mg/L. This observation indicated nitrifier inhibition due to aniline. In the continuous experiments, a hydraulic residence time of 8 to 24 hours and a solids retention time of 8 to 24 days were maintained. Complete nitrification took place at bulk aniline concentrations less than 0.5 mg/L, while at higher aniline concentrations, nitrification inhibition took place. The inhibitory effect of aniline on the nitrification process was modeled using uncompetitive inhibition kinetics. Modeling of batch processes yielded the value for the inhibition constant for aniline, Ki, as 3.3 mg/L. Modeling of continuous processes yielded criteria for stable process operation for nitrification under inhibitory conditions, which were also confirmed through experimental results. PMID- 12540094 TI - Giardia cysts in tertiary-treated wastewater effluents: are they infective? AB - The infectivity of Giardia lamblia cysts recovered in primary- and tertiary treated wastewater reclamation plant effluents was assessed in Mongolian gerbils (Meriones unguiculatus). Infections in gerbils inoculated with cysts from primary effluent concentrates demonstrated the presence of infectious G. lamblia cysts. No infectious cysts were detected by this method in concentrates of tertiary treated effluents. This study found that determination of cyst concentrations without viability or infectivity assessment may significantly overestimate the potential health risks associated with protozoan cysts in tertiary-treated wastewater effluents. PMID- 12540096 TI - Odor and volatile organic compound treatment by biotrickling filters: pilot-scale studies at hyperion treatment plant. AB - A pilot-scale biotrickling filter was installed at the Hyperion Treatment Plant in Los Angeles, California, to study hydrogen sulfide (odor) and volatile organic compound (VOC) removal from headworks waste air. The performance of the reactor was continuously monitored during a 10-month period. At an average empty bed gas residence time of 24 seconds, 10 to 50 ppm of hydrogen sulfide was consistently removed at greater than 98% efficiency, corresponding to an average volumetric elimination capacity of 5.2 g/m3 x h. Concentration profiles over the height of the reactor indicated nearly complete removal in the first section of the reactor, suggesting that elimination capacities up to 30 g/m3 x h could be obtained. The odor reduction (as dilution to threshold) was 98%, which correlated with the efficiency of removal of hydrogen sulfide as the primary pollutant. Volatile organic compounds were present at concentrations up to 225 ppb. Moderate but significant removal of toluene and benzene was observed when the biotrickling filter was operated with pH control to neutralize sulfuric acid production from hydrogen sulfide oxidation. Xylenes and chlorinated VOCs were not removed regardless of experimental conditions in the reactor. The results led to the conclusion that VOC removal is the limiting process in biotrickling filters for the simultaneous removal of hydrogen sulfide and VOCs at publicly owned treatment works. PMID- 12540095 TI - The sonolytic destruction of methyl tert-butyl ether present in contaminated groundwater. AB - Ultrasonic irradiation in the presence of ozone was used to efficiently eliminate methyl tert-butyl ether (MTBE) from groundwater. The sonolytic degradation of MTBE was investigated in three different reactor configurations and frequencies: vibrating-plate reactor (VPR, 358 kHz), near-field acoustical processor (NAP 20 and 16 kHz), and radial-tube resonator (RTR. 20 kHz). The sonochemical reactors can be ordered in terms of their efficiency with respect to the degradation of MTBE in the following way: VPR > RTR > NAP. The higher elimination rates of MTBE in groundwater by combined ultrasound-ozone systems are attributed to the effective conversion of ozone to the OH radical, even in the presence of high alkalinity. Carbonate radicals, which were formed from the oxidation of bicarbonate by hydroxyl radicals, are shown to react with MTBE via a hydrogen atom abstraction pathway. Methyl-tert-butyl ether was also rapidly eliminated from the groundwater underlying a major intemational airport by direct chemical oxidation with a mixture of hydrogen peroxide and ozone. PMID- 12540097 TI - An improved filter elution and cell culture assay procedure for evaluating public groundwater systems for culturable enteroviruses. AB - Large-scale virus studies of groundwater systems require practical and sensitive procedures for both sample processing and viral assay. Filter adsorption-elution procedures have traditionally been used to process large-volume water samples for viruses. In this study, five filter elution procedures using cartridge filters were evaluated for their effectiveness in processing samples. Of the five procedures tested, the third method, which incorporated two separate beef extract elutions (one being an overnight filter immersion in beef extract), recovered 95% of seeded poliovirus compared with recoveries of 36 to 70% for the other methods. For viral enumeration, an expanded roller bottle quantal assay was evaluated using seeded poliovirus. This cytopathic-based method was considerably more sensitive than the standard plaque assay method. The roller bottle system was more economical than the plaque assay for the evaluation of comparable samples. Using roller bottles required less time and manipulation than the plaque procedure and greatly facilitated the examination of large numbers of samples. The combination of the improved filter elution procedure and the roller bottle assay for viral analysis makes large-scale virus studies of groundwater systems practical. This procedure was subsequently field tested during a groundwater study in which large-volume samples (exceeding 800 L) were processed through the filters. PMID- 12540099 TI - Annual Session Proceedings. PMID- 12540098 TI - Ozonation of municipal wastewater effluents. AB - The increasing use of ozone in the treatment of municipal wastewater effluents has been stimulated by the need to achieve higher effluent quality and greater compliance with physicochemical and microbiological quality standards before discharge. These standards are applied when the effluent may pose a risk to the public through direct contact and where the effluent is used for agricultural purposes or water reclamation. Although various alternative technologies exist for upgrading wastewater effluents, ozone treatment may be the most appropriate approach in particular cases. This review summarizes the current status of the use of ozone for treating municipal effluents with respect to disinfection efficiency, its effect on the treatability of the effluent and on aggregate effluent parameters, the potential for the formation of ozonation byproducts, and its effect on the toxicity and mutagenicity of the effluent. The importance of treatment conditions (e.g., contact time) is also reviewed. PMID- 12540100 TI - "Intentional use of the Hawthorne effect to improve oral hygiene compliance in orthodontic patients". PMID- 12540101 TI - Development of a mission-focused faculty evaluation system. AB - The development of a new system for annual evaluation of faculty members is described. A narrative of the procedure, including accounts of the problems encountered, is used to show that such a process is too complex and too close to the fundamental identity of a dental school and the self-image of faculty members to be created in a one-time, rational effort or imposed by administrative edict. The process required five years to complete and involved an intermediate model. The goals of the new performance appraisal system were to minimize the extreme rating inflation and significant discrepancies from one chair rater to another that had existed previously. It was also a goal that the new system would create rich and effective feedback for faculty and would orient faculty members toward the mission of the school as a common focus. In achieving these goals, it is the authors' perception that faculty members at this dental school value procedural justice (fairness in the process), that evaluation is a political process, and that a performance appraisal system grounded in organizational mission rather than individual tasks of faculty members fits the emerging career model of knowledge professionals. PMID- 12540102 TI - U.S. predoctoral education in pediatric dentistry: its impact on access to dental care. AB - This study sought to identify faculty, organization, patient pool, and procedures taught in predoctoral pediatric dentistry programs using a questionnaire sent to all fifty-five U.S. dental schools in 2001. Forty-eight (87 percent) programs reported an average of 3.9 full-time and 2.1 part-time FTE faculty, resulting in a mean faculty to student ratio of 1:6.4. One-third employ general dentists to teach pediatric dentistry, and 36 percent report fewer faculty than five years ago. Two-thirds were stand-alone departments. Over half (55 percent) reported increases in patient pools, but also a lack of patients with restorative needs. Half of the programs supplemented school-based pools with special populations, and two-thirds sent students on external rotations, most often to treat high caries children. Those not using external rotations cited lack of faculty. Accepted patients averaged about four years, with only 6 percent of the pool under three years. Low-income or Medicaid-covered children accounted for 88 percent of school patient pools. Half of the schools felt the pool inadequate to meet competencies, attributable to lack of patients' restorative needs or inadequate intake numbers. Fewer than half of the programs (48 percent) provided hands-on experience with disabled patients, and one-third afforded every student with this experience. Pediatric dentistry was mentioned in fewer than half of the competency documents. Results suggest that U.S. pediatric dentistry predoctoral programs have faculty and patient pool limitations that affect competency achievement and adversely affect training and practice. PMID- 12540103 TI - Issues regarding practical validity and gender bias of the Perceptual Abilities Test (PAT). AB - A brief history of the Dental Admission Test is provided, with emphasis on the development and utilization of the Perceptual Abilities Test (PAT). Concerns regarding the predictive utility of the PAT are discussed, and alternative approaches to assessing perceptual and motor skills are examined. To assess the predictive validity of the PAT, scores were collected for 492 students who graduated from the University of Oklahoma College of Dentistry over a ten-year period. Scores were compared to Preclinical Operative Dentistry practical exam scores for each subject. Correlational analysis indicated the PAT maintains a statistically significant capacity for predicting technical performance in dental school, but the practical significance of the predictive capacity is limited. The results also indicate a gender bias favoring male applicants. Implications regarding continued reliance by admissions committees on the Perceptual Abilities Test are discussed and recommendations are provided, including modification of the PAT, use of the PAT only as a gross screening tool, and discontinuing the use of the PAT in making dental school admission decisions. PMID- 12540104 TI - Evaluation of a clinical outcomes assessment tool in a U.S. dental school. AB - Quality assurance (QA) programs in dental schools have a component of their program devoted to treatment outcomes. To this end, our institution has implemented TOUCH (Treatment Outcomes Unacceptable for Clinical Health) seminars and Unusual Occurrence Reports (UORs). The seminars allow a faculty member to present a case to faculty and students with feedback from the audience on how the case was managed. The UORs track clinical incidents outside the range of normal. Participation in both of these QA measures has been less than expected. The goal of the current study was to discover the reasons for participation and lack of participation. A twelve-item survey was completed by seventy-one clinical faculty members and analyzed for trends. Faculty report only 28.3 percent of the unacceptable outcome cases they know about. The two most common reasons given for reporting an unusual occurrence were that it would help the institution reduce similar incidents and it would provide an opportunity to share learning experiences. The most common reason given for not reporting an unusual occurrence was not remembering to do so. Faculty members were most willing to present a TOUCH seminar if guaranteed that no negative repercussions would result. Suggestions for increasing participation in both programs include emphasizing their value, modifying the seminar format, providing more reminders, and reassuring against repercussions. PMID- 12540105 TI - Assessing the effectiveness of a new curriculum: Part I. AB - Although it is important to assess the effectiveness of programs, courses, and teaching methods to ensure that goals are being achieved, it is very difficult to evaluate the impact of fundamental changes in a whole curriculum. This paper reviews measures that have been used in the past in dentistry and medicine for evaluating academic programs: curriculum guidelines; competency documents; discussion and focus groups; competency examinations; board examinations; oral comprehensive examinations; student, alumni, and patient satisfaction surveys; evaluation by instructors; and clinical productivity. We conclude that, since no standard method exists, several tools should be used to obtain a multidimensional assessment. PMID- 12540106 TI - Curriculum evaluation of ethical reasoning and professional responsibility. AB - This exploratory study evaluated curricular content and evaluation mechanisms related to ethics and professionalism in the baccalaureate dental hygiene program at Idaho State University. Competency-based education requires enhanced student preparation in ethical reasoning, critical thinking, and decision-making. Graduates must integrate concepts, beliefs, principles, and values to fulfill ethical and professional responsibilities. Methods included 1) development of five supporting competencies defining ethics and professionalism to provide a framework for curricular evaluation; 2) assessment of all course content and evaluation methods for each supporting competency; 3) evaluation of students' clinical performance based on professional judgment grades; and 4) survey of junior (n=30) and senior (n=27) students' attitudes about dental hygiene practice related to ethics and professionalism. Results revealed that most courses include content and evaluation related to at least one supporting competency; however, authentic evaluation is weak. Clinical instructors rarely relate evaluations to ethical principles or values. Surveys showed significant differences between junior and senior students' attitudes about ethics and professionalism in six of thirty-four areas (the six were laws and regulations; communication and interpersonal skills; problem solving; professional activities/programs; integrity; and safe work environment). This article shares one approach for evaluating curricular content and evaluation methods designed to develop student competence in ethical reasoning and professionalism. Based upon the study's findings, recommendations are made for curricular enhancement via authentic evaluation and faculty training. PMID- 12540107 TI - Therapeutic use of hyperbaric oxygen for irradiated dental implant patients: a systematic review. AB - The aim of this systematic review was to investigate the effectiveness of hyperbaric oxygen (HBO) therapy for irradiated patients who require dental implants using data from randomized controlled clinical trials (RCTs). The review was prepared according to Cochrane Collaboration guidelines. The Cochrane Oral Health Group Specialist Register and the Cochrane Controlled Trials Register were searched (Cochrane Library 2002, Issue 2), together with Medline from 1966 or Embase from 1974. Several journals were hand-searched, and fifty-five implant manufacturers were contacted in an attempt to identify ongoing or unpublished studies. The results were that no RCTs comparing HBO with no HBO for implant treatment in irradiated patients were identified. Our principal conclusions are that clinicians ought to be aware and make patients aware of the lack of reliable clinical evidence for or against the clinical effectiveness of HBO therapy in irradiated patients requiring dental implants. There is a need for RCTs to determine the effectiveness of HBO. PMID- 12540108 TI - Dental education in Europe: the challenges of variety. AB - Dental education varies considerably across Europe, with differing traditions of stomatology (dentistry as a specialty of medicine) and odontology (single autonomous discipline). Dental curricula within the European Union (EU) are governed by European law expressed in directives that are binding on all EU member states. The Dental Directives (78/686/EC) base the curriculum on the odontological model, but compliance by individual schools is often poor. The differences within the EU will likely intensify with the accession of Eastern/Central European countries where the stomatological tradition is strong. Moreover, current proposals within the EU will reduce even the limited existing effectiveness of the Dental Directives. The DentEd Thematic Network Project, which aims to promote convergence of European curricula through voluntary self assessment and outside peer review, has involved about 25 percent of European schools. Its effectiveness in inducing changes in individual schools is unknown. It is not an accreditation system, and there is no intention to establish a European-wide common curriculum. Dentists' vocational training, here defined as "the organised education of the newly qualified dentist in supervised practice," is present in various models in many European countries, but is compulsory in only a few. Continuing dental education (CDE) is encouraged in most countries, but CDE-dependent licensure is required in only two. PMID- 12540109 TI - Advancement of women in dental education: trends and strategies. PMID- 12540110 TI - Where have all the mentors gone? PMID- 12540111 TI - Technique sensitivity in bonding to vital, acid-etched dentin. AB - Just as vital dentin is moist after removing the smear layer, avoiding collapse of the collagen matrix after acid-etching requires in vivo validation. This study hypothesizes that there is no difference between moist bonding performed in vitro or in vivo, and that excessive drying or wetting of vital acid-etched dentin produces inferior results. Resin-dentin interfaces bonded with a moist bonding technique (control), either in vitro or in vivo with Excite DSC (Vivadent), were examined with and without tracer penetration using transmission electron microscopy. Specimens bonded in vivo under excessively dry and wet conditions were also examined. The patterns of silver deposition were similar within the adhesive and hybrid layers created in vitro or in vivo. No hybrid layer was observed in vivo after excessive drying. Excessive wetting in vivo resulted in more extensive nanoleakage and water tree formation along resin-dentin interfaces. PMID- 12540112 TI - Voids and porosities in class I micropreparations filled with various resin composites. AB - In this in vitro study, voids inside a minimal occlusal restoration using different consistencies of resin composite and various application techniques were investigated. One hundred and fifty-two simulated, minimally invasive preparations, including a prepared fissure and an excavated carious dentin lesion, were ground in perspex blocks. After applying an acrylic primer (Artglass Connector, Kulzer), the preparations were restored with the adhesive PhotoBond (Kuraray) and one of three resin composites: a packable composite (Prodigy Condensable-Kerr), a syringable composite (Clearfil Photo Posterior, Kuraray) and a flowable composite (Revolution, Kerr). The restorations were inserted according to eight protocols (n = 19). In three groups, the composite was placed in bulk. In another three groups, a layer of flowable composite was placed first, then cured, followed by a second layer of one of the three composites. In two groups, the first layer of flowable composite was left uncured before a second layer of a packable or syringable composite was inserted. The perspex blocks were sectioned and inspected for the presence of voids. Statistical analysis was conducted using Fischer's exact tests at p < 0.05. The results showed that restoring minimal preparations in the absence of porosities and voids was very difficult to achieve. Placing a layer of flowable composite that was left uncured, directly followed by injecting a medium-viscous composite, was the technique that resulted in the most homogeneous restoration. PMID- 12540114 TI - Effects of regional enamel and prism orientation on resin bonding. AB - Human enamel, with its prismatic, rod-like apatitic morphology, is an anisotropic material. Because of this structural anisotropy, variation in enamel bonding sites might influence the bonding ability of current adhesive systems. This study investigated the effects of regional enamel and the direction of enamel sectioning on the bonding ability of two commercially available resin adhesives: a self-etching primer system (Clearfil SE Bond) and a one-bottle adhesive system intended for use with a total-etch wet bonding technique (Single Bond). Two regions of enamel, cuspal and mid-coronal enamel, were chosen, then sectioned in three different directions, horizontally, axially and tangentially. Slices of the sectioned enamel were then bonded with each adhesive system and submitted to a micro-shear bond test. The results of a micro-shear bond testing showed that the bonding of a one-bottle adhesive system (Single Bond) to enamel was high at the surface perpendicular to the enamel prisms (40 MPa to 51 MPa) and low at the surface parallel to the enamel prisms (24 MPa to 27 MPa). In the case of a self etching primer system (Clearfil SE Bond), 35 MPa to 45 MPa bond strengths were obtained from all surfaces. The bond strengths of the two adhesive systems were significantly influenced by the anisotropic structure of enamel (p < 0.05). However, the effect of a self-etching primer system was less influenced by the orientation of the prismatic structure of enamel than that of a one-bottle adhesive system (p < 0.05). SEM and CLSM microphotographs showed that the self etching primer effectively modified the smear layer without being excessively destructive to the enamel. PMID- 12540113 TI - Effects of in-office bleaching products on surface finish of tooth-colored restorations. AB - A number of "high power" in-office bleaching products have recently been re introduced into the market. The use of such strong oxidizing agents has raised questions as to possible adverse effects on tooth structure and restorative materials. This study evaluated the effects of 35% carbamide peroxide (Opalescence Quick) and 35% hydrogen peroxide (Opalescence Xtra) on the surface finish of four tooth-colored restorative materials (Spectrum TPH, Dyract AP, Reactmer and Fuji II LC). Twenty-seven matrix-finished specimens of each material were fabricated, stored in distilled water at 37 degrees C for seven days and randomly divided into three groups. Specimens in Group 1 were stored in distilled water at 37 degrees C (control). Specimens in Groups 2 and 3 were treated with 35% carbamide peroxide and 35% hydrogen peroxide, respectively. A total of three 30-minute bleaching sessions were conducted at one-week intervals. Storage medium during the hiatus period was distilled water at 37 degrees C. Surface roughness measurements were carried out using profilometry after each bleaching session. Data was analyzed using ANOVA/Scheffe's test at a 0.05 significance level. No significant difference in surface roughness was observed between the bleached and the control groups for all materials. In-office bleaching products are not detrimental to the surface finish of composites, compomers, giomers and resin modified glass ionomer cements. PMID- 12540115 TI - The effect of a "resin coating" on the interfacial adaptation of composite inlays. AB - The relatively low bond strengths of resin cements to dentin may result in poor interfacial adaptation of composite inlays. This study determined whether the interfacial adaptation of composite inlays could be improved by applying an adhesive system and a low viscosity microfilled resin to the prepared cavity walls before making an impression. Ten MOD cavities were prepared on extracted human premolars with gingival margins located above and below the cemento-enamel junction. A "resin coat" consisting of a self-etching primer system (Clearfil SE Bond) and a low viscosity microfilled resin (Protect Liner F) was applied to the cavities of half of the prepared teeth, while the remaining teeth served as non coated control specimens. All the teeth were restored with composite inlays (Estenia) fabricated by the indirect method and cemented with a dual-cured resin cement (Panavia F). After finishing the margins with superfine burs, the bonded inlays were thermocycled between 4 degrees C and 55 degrees C for 400 cycles. Specimens were sectioned with a diamond saw and the tooth-restoration interfaces were observed with a confocal laser scanning microscope. The data were analyzed with two-way ANOVA and Fisher's PLSD test (p < 0.05). The percentage length of gap formation at the dentin-restoration interface of the "resin coated" teeth (7.1 +/- 3.5) was significantly less than that of the non-coated teeth (85.7 +/- 6.7) (p < 0.05). The concept of coating the prepared cavity with an adhesive system and a low viscosity microfilled resin resulted in observing fewer gaps at the internal dentin-restoration interface compared with the non-coated specimens. PMID- 12540116 TI - Effects of finishing/polishing techniques on microleakage of resin-modified glass ilonomer cement restorations. AB - This study investigated the effect of finishing/polishing techniques on the microleakage of resin-modified glass ionomer restorations. Class V preparations were made on the buccal and lingual/palatal surfaces of freshly extracted teeth. The cavities on each tooth were restored with Fuji II LC (FT [GC]) and Photac-Fil Quick (PF [3M-ESPE]) according to manufacturers' instructions. Immediately after light-polymerization, gross finishing was done with eight-fluted tungsten carbide burs. The teeth were then randomly divided into four groups and finishing/polishing was done with one of the following systems: (a) Robot Carbides (RC); (b) Super-Snap system (SS); (c) OneGloss (OG) and (d) CompoSite Polishers (CS). The sample size for each material-finishing/polishing system combination was eight. After finishing/polishing, the teeth were stored in distilled water at 37 degrees C for one week. The root apices were then sealed with acrylic and two coats of varnish was applied 1 mm beyond the restoration margins. The teeth were subsequently subjected to dye penetration testing (0.5% basic fuchsin), sectioned and scored. Data was analyzed using Kruskal-Wallis and Mann-Whitney U tests at a significance level of 0.05. Results of statistical analysis were as follows: Enamel margins: PF-OG 12 mg/dl). To determine the cause of the hypercalcemia, additional diagnostic tests, including thoracic and abdominal radiographs and a parathyroid panel (parathyroid hormone (PTH), ionized calcium, and parathyroid hormone-related protein (PTH-rp) assays), were performed. The radiographs did not reveal lesions suggestive of neoplasia. A parathyroid panel was obtained twice. Both times the PTH (23.4 and 46.4 pmol/L, normal = 2.91 to 4.57 pmol/L) and ionized calcium (1.68 and 2.10 mmol/L, normal = 1.31 to 1.37 mmol/L) were increased above values for adult females with normal calcium concentration. A tentative diagnosis of primary hyperparathyroidism was made. After a gamma radiation scan and magnetic resonance imaging of the neck were done, exploratory surgery was performed to identify and remove the affected gland. After gland removal, the baboon's serum calcium, PTH (1.6 pmol/L), and ionized calcium (1.59 mmol/L) values decreased. Results of histologic examination confirmed the diagnosis of benign solitary parathyroid adenoma. PMID- 12540173 TI - Epididymal cribriform hyperplasia with nuclear atypia in p53 homozygous knockout mice on a mixed 129/Sv-FVB/N background. AB - Epididymal cribriform hyperplasia (ECH) is a variant of normal epididymal histologic features in men, and has also been reported in rats, mice, dogs, cats, and bulls. The epididymal change has been associated with aging, testicular atrophy, cryptorchidism, and germ cell tumors. Epididymal cribriform hyperplasia was observed in p53 homozygous knockout mice on a mixed 129/Sv-FVB/N background, but not in wild-type or heterozygous mice. The aim of the study reported here was to determine the prevalence and characterize the morphologic, immunohistochemical, and ultrastructural features of ECH in these mice. Epididymal cribriform hyperplasia was present in 88% (72/82) of male mice ranging in age from seven to 65 weeks. The lesion was characterized microscopically by epithelial cells with atypical hyperchromatic nuclei, vacuolization, intratubular lumina formation, infrequent apoptosis, and rare mitotic figures. In contrast to germ cells, the cells of ECH did not express alpha-fetoprotein, carcinoembryonic antigen, or S-100. Ultrastructurally, the cells were pleomorphic with stereocilia at their apical borders and within intratubular lumina, and were supported by a basement membrane. Although 14% (10/72) of mice had concomitant testicular neoplasia, ECH did not appear to be a preneoplastic change. Investigators using these mice for modeling human disease should be aware of the background prevalence of this lesion. PMID- 12540174 TI - Antagonism of Helicobacter pylori by bacteriocins of lactic acid bacteria. AB - Antimicrobial activity of seven bacteriocins produced by lactic acid bacteria against Helicobacter pylori strains (ATCC 43504, Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH [DSM] 4867, DSM 9691, and DSM 10242) was investigated in vitro using a broth microdilution assay. The bacteriocins chosen for the study were nisin A; lacticins A164, BH5, JW3, and NK24; pediocin PO2; and leucocin K. Antimicrobial activity of the bacteriocins varied among the H. pylori strains tested, of which strain ATCC 43504 was the most tolerant. Among the bacteriocins tested, lacticins A164 and BH5 produced by Lactococcus lactis subsp. lactis A164 and L. lactis BH5, respectively, showed the strongest antibacterial activity against H. pylori strains. MICs of the lacticins against H. pylori strains, when assessed by the critical dilution micromethod, ranged from 0.097 to 0.390 mg/liter (DSM strains) or from 12.5 to 25 mg/liter (ATCC 43504), supporting the strain-dependent sensitivity of the pathogen. Pediocin PO2 was less active than the lacticins against four strains of H. pylori, and leucocin K was the least active peptide, with no inhibition toward H. pylori ATCC 43504. Anti Helicobacter activity of lacticin A164 was dependent on initial inoculum size as well as concentration of the bacteriocin added. PMID- 12540175 TI - Alfalfa sprouts and Salmonella Kottbus infection: a multistate outbreak following inadequate seed disinfection with heat and chlorine. AB - Raw sprouts have been implicated in a number of foodborne disease outbreaks. Because contaminated seeds are usually responsible, many sprout producers attempt to disinfect seeds before germination and detect sprout contamination during production. In March 2001, we detected an increased number of Salmonella serotype Kottbus isolates in California. Overall, we identified 31 cases from three western states. To identify the cause, we conducted a case-control study with the first 10 identified case-patients matched to 20 controls by age, sex, and residential area. Our case-control study found illness to be statistically associated with alfalfa sprout consumption. The traceback investigation implicated a single sprouter, where environmental studies yielded Salmonella Kottbus from ungerminated seeds and floor drains within the production facility. Pulsed-field gel electrophoresis patterns of all patient, seed, and floor drain Salmonella Kottbus isolates were indistinguishable. Most implicated sprouts were from seeds that underwent heat treatment and soaking with a 2,000-ppm sodium hypochlorite solution rather than the Food and Drug Administration (FDA) recommended 20,000-ppm calcium hypochlorite soak. Other implicated seeds had been soaked in a calcium hypochlorite solution that, when tested, measured only 11,000 ppm. The outbreak might have been averted when screening tests of sprout irrigation water detected Salmonella in January; however, confirmatory testing of these samples was negative (but testing improperly utilized refrigerated irrigation water). Producers should use the enrichment broth of positive screening samples, not refrigerated irrigation water, for confirmatory testing. Until other effective disinfection technologies are developed, producers should adhere to FDA recommendations for sprout seed disinfection. PMID- 12540176 TI - Evaluation of methods to improve detection of Escherichia coli O157:H7 in fresh produce by multiplex polymerase chain reaction. AB - Multiplex polymerase chain reaction (PCR) analysis was used to detect two genes encoding Shiga-like toxins (stx1 and stx2) and a universal Escherichia coli gene (gadA/B) in fresh produce spiked with E. coli O157:H7. Current U.S. Food and Drug Administration procedures for the analysis of fresh produce include the use of the rinsate from an initial rinse for the analysis of several potential pathogens, including E. coli O157:H7. In this study, several procedures were evaluated for their ability to increase the sensitivity of PCR analysis of rinsates from 15 types of produce. The procedures evaluated included the preliminary clarification and concentration of templates by centrifugation and the treatment of templates with compounds reported to facilitate nucleic acid amplification, including polyvinlypolypyrrolidone (PVPP), nonfat dry milk (NFDM), and InstaGene. The preliminary concentration of rinsates resulted in moderate improvements in detection sensitivity. The use of PVPP-treated templates in PCR reaction mixtures did not further improve sensitivity, but the inclusion of NFDM treated templates increased sensitivity by an order of magnitude for 12 rinsates. The incorporation of InstaGene also improved the detection capability of the analysis; this procedure yielded the strongest gel bands for eight rinsates. However, for four other rinsates, the use of this reagent decreased sensitivity; these four rinsates were those for the produce varieties with the largest surface areas and were the most turbid rinsates. The use of facilitating compounds to block PCR inhibition may enable an analysis for Shiga toxin-producing E. coli in fresh produce to be completed in 1 to 2 days, rather than the 5 days required for current methods. PMID- 12540177 TI - Fate of Escherichia coli O157:H7 during composting of bovine manure in a laboratory-scale bioreactor. AB - Inactivation profiles of Escherichia coli O157:H7 in inoculated bovine manure based compost ingredients were determined by composting these ingredients in a bioreactor under controlled conditions. A 15-liter bioreactor was constructed to determine the fate of E. coli O157:H7 and changes in pH, moisture content, temperature, and aerobic mesophilic and thermophilic bacterial counts during composting. Fresh cow manure, wheat straw, cottonseed meal, and ammonium sulfate were combined to obtain a moisture content of ca. 60% and a carbon/nitrogen ratio of 29:1. The compost ingredients were held in the bioreactor at a constant external temperature of 21 or 50 degrees C. Self-heating of the ingredients due to microbial activity occurred during composting, with stratified temperatures occurring within the bioreactor. At an external temperature of 21 degrees C, self heating occurred for 0 to 3 days, depending on the location within the bioreactor. E. coli O157:H7 populations increased by 1 to 2 log10 CFU/g during the initial 24 h of composting and decreased by ca. 3.5 log10 CFU/g near the bottom of the bioreactor and by ca. 2 log10 CFU/g near the middle and at the top during 36 days of composting. At an external temperature of 50 degrees C. E. coli O157:H7 was inactivated rapidly (by ca. 4.9 log10 CFU/g at the top of the bioreactor, by 4.0 log10 CFU/g near the middle, and by 5.9 log10 CFU/g near the bottom) within 24 h of composting. When inoculated at an initial level of ca. 10(7) CFU/g. E. coli O157:H7 survived for 7 days but not for 14 days at all three sampling locations, as indicated by either direct plating or enrichment culture. At the top of the bioreactor a relatively constant moisture content of 60% was maintained, whereas the moisture content near the bottom decreased steadily to 37 to 45% over 14 days of composting. The pH of the composting mixture decreased to ca. 6 within 1 to 3 days and subsequently increased to 8 to 9. Results obtained in this study indicate that large populations (10(4) to 10(7) CFU/g) of E coli O157:H7 survived for 36 days during composting in a bioreactor at an external temperature of 21 degrees C but were inactivated to undetectable levels after 7 to 14 days when the external temperature of the bioreactor was 50 degrees C. Hence, manure contaminated with large populations (e.g., 10(7) CFU/g) of E. coli O157:H7 should be composted for more than 1 week, and preferably for 2 weeks, when held at a minimum temperature of 50 degrees C. PMID- 12540178 TI - Comparison of sublethal injury induced in Salmonella enterica serovar Typhimurium by heat and by different nonthermal treatments. AB - We have studied sublethal injury in Salmonella enterica serovar Typhimurium caused by mild heat and by different emerging nonthermal food preservation treatments, i.e., high-pressure homogenization, high hydrostatic pressure, pulsed white light, and pulsed electric field. Sublethal injury was determined by plating on different selective media, i.e., tryptic soy agar (TSA) plus 3% NaCl, TSA adjusted to pH 5.5, and violet red bile glucose agar. For each inactivation technique, at least five treatments using different doses were applied in order to cover an inactivation range of 0 to 5 log units. For all of the treatments performed with a technique, the logarithm of the viability reductions measured on each of the selective plating media was plotted against the logarithm of the viability reduction on TSA as a nonselective medium, and these points were fined by a straight line. Sublethal injury between different techniques was then compared by the slope and the y intercept of these regression lines. The highest levels of sublethal injury were observed for the heat and high hydrostatic pressure treatments. Sublethal injury after those treatments was observed on all selective plating media. For the heat treatment, but not for the high-pressure treatment, sublethal injury occurred at low doses, which were not yet lethal. The other nonthermal techniques resulted in sublethal injury on only some of the selective plating media, and the levels of injury were much lower. The different manifestations of sublethal injury were attributed to different inactivation mechanisms by each of the techniques, and a mechanistic model is proposed to explain these differences. PMID- 12540179 TI - Use of diacetyl to reduce the load of Vibrio vulnificus in the Eastern oyster, Crassostrea virginica. AB - Vibrio vulnificus is a highly virulent human pathogen that occurs naturally among the microflora of oysters. This organism has two portals of entry into humans, one of which is ingestion. Oysters containing V. vulnificus consumed in a raw or undercooked state often serve as a vehicle for the transmission of this organism. Previous studies conducted in our laboratory have examined various generally recognized as safe compounds and have determined that diacetyl, a component of butter, is among the most effective of these compounds in reducing loads of V. vulnificus in oysters. The purpose of this study was to further examine the role of diacetyl, along with that of depuration, in reducing loads of V. vulnificus. Shellstock oysters were treated with various concentrations of diacetyl, and we found that many of the oysters ceased pumping when diacetyl was added. The data obtained in this study indicated that treatment with diacetyl is ineffective; however, any reduction in V. vulnificus numbers may be masked when groups of oysters, some of which may not have taken up diacetyl, are sampled. We then investigated the efficacy of diacetyl in lowering levels of V. vulnificus in shucked oysters. Diacetyl was found to significantly reduce the load of V. vulnificus in shucked oysters containing natural populations. Overall, it appears that treatment with diacetyl is ineffective for shellstock oysters, although it has potential for use in reducing loads of V. vulnificus in shucked oysters. PMID- 12540180 TI - Efficacy of ozone in killing Listeria monocytogenes on alfalfa seeds and sprouts and effects on sensory quality of sprouts. AB - A study was done to determine the efficacy of aqueous ozone treatment in killing Listeria monocytogenes on inoculated alfalfa seeds and sprouts. Reductions in populations of naturally occurring aerobic microorganisms on sprouts and changes in the sensory quality of sprouts were also determined. The treatment (10 or 20 min) of seeds in water (4 degrees C) containing an initial concentration of 21.8 +/- 0.1 microg/ml of ozone failed to cause a significant (P < or = 0.05) reduction in populations of L. monocytogenes. The continuous sparging of seeds with ozonated water (initial ozone concentration of 21.3 +/- 0.2 microg/ml) for 20 min significantly reduced the population by 1.48 log10 CFU/g. The treatment (2 min) of inoculated alfalfa sprouts with water containing 5.0 +/- 0.5, 9.0 +/- 0.5, or 23.2 +/- 1.6 microg/ml of ozone resulted in significant (P < or = 0.05) reductions of 0.78, 0.81, and 0.91 log10 CFU/g, respectively, compared to populations detected on sprouts treated with water. Treatments (2 min) with up to 23.3 +/- 1.6 microg/ml of ozone did not significantly (P > 0.05) reduce populations of aerobic naturally occurring microorganisms. The continuous sparging of sprouts with ozonated water for 5 to 20 min caused significant reductions in L. monocytogenes and natural microbiota compared to soaking in water (control) but did not enhance the lethality compared to the sprouts not treated with continuous sparging. The treatment of sprouts with ozonated water (20.0 microg/ml) for 5 or 10 min caused a significant deterioration in the sensory quality during subsequent storage at 4 degrees C for 7 to 11 days. Scanning electron microscopy of uninoculated alfalfa seeds and sprouts showed physical damage, fungal and bacterial growth, and biofilm formation that provide evidence of factors contributing to the difficulty of killing microorganisms by treatment with ozone and other sanitizers. PMID- 12540181 TI - Listeria monocytogenes contamination patterns for the smoked fish processing environment and for raw fish. AB - Reliable data on the sources of Listeria monocytogenes contamination in cold smoked fish processing are crucial in designing effective intervention strategies. Environmental samples (n = 512) and raw fish samples (n = 315) from two smoked fish processing facilities were screened for L. monocytogenes, and all isolates were subtyped by automated ribotyping to examine the relationship between L. monocytogenes contamination from raw materials and that from environmental sites. Samples were collected over two 8-week periods in early spring and summer. The five types of raw fish tested included lake whitefish, sablefish, farm-raised Norwegian salmon, farm-raised Chilean salmon, and feral (wild-caught) salmon from the U.S. West Coast. One hundred fifteen environmental samples and 46 raw fish samples tested positive for L. monocytogenes. Prevalence values for environmental samples varied significantly (P < 0.0001) between the two plants; plant A had a prevalence value of 43.8% (112 of 256 samples), and plant B had a value of 1.2% (3 of 256 samples). For plant A, 62.5% of drain samples tested positive for L. monocytogenes, compared with 32.3% of samples collected from other environmental sites and 3.1% of samples collected from food contact surfaces. Ribotyping identified 11 subtypes present in the plant environments. Multiple subtypes, including four subtypes not found on any raw fish, were found to persist in plant A throughout the study. Contamination prevalence values for raw fish varied from 3.6% (sablefish) to 29.5% (U.S. West Coast salmon), with an average overall prevalence of 14.6%. Sixteen separate L. monocytogenes subtypes were present on raw fish, including nine that were not found in the plant environment. Our results indicate a disparity between the subtypes found on raw fish and those found in the processing environment. We thus conclude that environmental contamination is largely separate from that of incoming raw materials and includes strains persisting, possibly for years, within the plant. Operational and sanitation procedures appear to have a significant impact on environmental contamination, with both plants having similar prevalence values for raw materials but disparate contamination prevalence values for the environmental sites. We also conclude that regular L. monocyrogenes testing of drains, combined with molecular subtyping of the isolates obtained, allows for efficient monitoring of persistent L. monocytogenes contamination in a processing plant. PMID- 12540182 TI - Effectiveness of trisodium phosphate against Listeria monocytogenes on excised and nonexcised chicken skin. AB - The influence of sample type (i.e., excised versus nonexcised chicken skin) on the efficiency of trisodium phosphate (TSP) solutions in reducing Listeria monocytogenes populations and inhibiting their growth during refrigerated storage was studied. Whole chicken legs and excised chicken leg skin fragments inoculated with 10(8) CFU of L. monocytogenes per ml were dipped for 15 min in sterile tap water (control) or in a solution containing 8, 10, or 12% TSP. L. monocytogenes counts were determined after 0, 1, 3, and 5 days of refrigerated storage (2 degrees C). The decontamination effect of TSP was greater for excised skin than for whole legs. Microbial differences between control and TSP-treated samples were significantly larger for excised skin than for whole legs for 9 (75%) of 12 tested combinations of TSP concentrations and storage times. These differences varied from 1.05 +/- 0.26 log10 cycles (day 1) to 3.30 +/- 0.14 log10 cycles (day 5) for nonexcised-skin samples (whole legs) and from 1.54 +/- 0.48 log10 cycles (day 1) to 4.28 +/- 0.86 log10 cycles (day 5) for excised-skin samples. Significantly larger reductions were observed from the third day of refrigerated storage onward. The TSP concentration was a significant factor in the reduction of L. monocytogenes populations. These results suggest that bacteria are more readily accessible to TSP in excised than in nonexcised chicken skin and that the type of sample used to ascertain the efficacy of antimicrobial surface treatments may influence the findings of this type of study. PMID- 12540183 TI - Comparison of different most-probable-number methods for enumeration of Listeria in poultry. AB - To estimate levels of Listeria spp. in poultry and to select the most appropriate enumeration method for routine analysis, 40 naturally contaminated retail chicken carcasses were tested in Ponferrada (Leon, N.W. Spain) using the direct plate count technique and various most-probable-number (MPN) designs (UVM I [University of Vermont modified Listeria enrichment broth], Fraser enrichment broth, or both were used in 3-, 5-, and 10-tube MPN techniques). MPN estimation was obtained from the number of tubes with Listeria confirmed (after streaking on PALCAM and modified Oxford agars: "true" MPN) and from the number of dark Fraser broth tubes ("predictive" MPN). Samples were analyzed in duplicate. Low levels of Listeria were found (< 110 CFU/g). The direct plate count technique was totally ineffective for enumerating Listeria in poultry. The single-step (UVM I) and the two-step (UVM I-Fraser) MPN methods gave comparable estimations and a low number of significantly discrepant predictions. Using a single-step method with Fraser broth, lower true MPNs were obtained. The number of tubes used (3, 5, or 10) did not have a substantial influence on the results. Similar estimations, highly correlated (r = 0.538 to 0.968; P < 0.001), were found with (true MPN) and without (predictive MPN) plating confirmation when using the two-step MPN method. The statistical evaluation of the differential character of Fraser broth as part of the two-step MPN method showed high sensitivity (87.5 to 92.5%), specificity (95.2 to 98.6%), efficiency (94.2 to 97.6%), and predictive values (73.6 to 89.9% for a positive test and 98.0 to 98.9% for a negative test). Taking into account these results, we suggest the convenience of using a 3- or 5-tube two-step (UVM I Fraser) MPN method with estimations obtained from the number of tubes with darkening, without confirmation, in order to achieve great savings in time and money. PMID- 12540184 TI - Incidence of Clostridium perfringens in commercially produced cured raw meat product mixtures and behavior in cooked products during chilling and refrigerated storage. AB - A total of 445 whole-muscle and ground or emulsified raw pork, beef, and chicken product mixtures acquired from industry sources were monitored over a 10-month period for vegetative and spore forms of Clostridium perfringens. Black colonies that formed on Shahidi-Ferguson perfringens (SFP) agar after 24 h at 37 degrees C were considered presumptive positive. Samples that were positive after a 15-min heat shock at 75 degrees C were considered presumptive positive for spores. Of 194 cured whole-muscle samples, 1.6% were positive; spores were not detected from those samples. Populations of vegetative cells did not exceed 1.70 log10 CFU/g and averaged 1.56 log10 CFU/g. Of 152 cured ground or emulsified samples, 48.7% were positive, and 5.3% were positive for spores. Populations of vegetative cells did not exceed 2.72 log10 CFU/g and averaged 1.98 log10 CFU/g; spores did not exceed 2.00 log10 CFU/g and averaged 1.56 log10 CFU/g. Raw bologna (70% chicken), chunked ham with emulsion, and whole-muscle ham product mixtures were inoculated with C. perfringens spores (ATCC 12916, ATCC 3624, FD1041, and two product isolates) to ca. 3.0 log10 CFU/g before being subjected either to thermal processes mimicking cooking and chilling regimes determined by in-plant temperature probing or to cooking and extended chilling regimes. Populations of C. perfringens were recovered on SFP from each product at the peak cook temperatures, at 54.4, 26.7, and 7.2 degrees C, and after up to 14 days of storage under vacuum at 4.4 degrees C. In each product, populations remained relatively unchanged during chilling from 54.4 to 7.2 degrees C and declined slightly during refrigerated storage. These findings indicate processed meat products cured with sodium nitrite are not at risk for the growth of C. perfringens during extended chilling and cold storage. PMID- 12540185 TI - On-farm and postharvest processing sources of bacterial contamination to melon rinds. AB - Multistate and international foodborne illness outbreaks, particularly involving cantaloupe and often involving rare Salmonella spp., have increased dramatically over the past 13 years. This study assessed the sources and extent of melon rind contamination in production fields and at processing and packing facilities. In the spring of 1999, cantaloupe (Cucumis melo L. [reticulatus group] cv. Cruiser) sampled from two sites in the Rio Grande River Valley showed that postharvest processed melon rinds often had greater plate counts of bacterial contaminants than field-fresh melons. Cantaloupe in the field had 2.5 to 3.5 log CFU g(-1) rind total coliforms by aerobic plate counts, whereas washed melons had 4.0 to 5.0 log CFU g(-1). In the fall of 1999, coliforms on honeydew melons (C. melo [inodorous group] cv. Honey Brew) ranged from 2.6 to 3.7 log CFU g(-1) after processing, and total and fecal coliforms and enterococci never fell below 2.5 log CFU g(-1). A hydrocooler at another site contaminated cantaloupe rinds with up to 3.4 log CFU g(-1) total and fecal enterococci; a secondary rinse with chlorinated water incompletely removed these bacteria. Sources of coliforms and enterococci were at high levels in melon production soils, especially in furrows that were flood irrigated, in standing water at one field, and in irrigation water at both sites. At one processing facility, wash water pumped from the Rio Grande River may not have been sufficiently disinfected prior to use. Because soil, irrigation water, and process water were potential sources of bacterial contamination, monitoring and management on-farm and at processing and packing facilities should focus on water quality as an important control point for growers and packers to reduce bacterial contamination on melon rinds. PMID- 12540186 TI - Male-specific coliphages as an additional fecal contamination indicator for screening fresh carrots. AB - The objective of this study was to evaluate the efficacy of male-specific (F+) coliphages as a fecal-contamination indicator for fresh carrots. The prevalence of specific pathogens and indicator organisms on the surface of carrots obtained from a farm, truck, and processing shed was studied. Twenty-five carrot samples collected from each of these locations were washed, and aliquots of the wash were analyzed for the presence of F+ coliphages, Escherichia coli, Salmonella, and Shigella. Additionally, the Salmonella isolates were genotyped using pulsed-field gel electrophoresis (PFGE). Our studies detected the presence of F+ coliphages, E. coli, and Salmonella on carrots. All samples, however, tested negative for Shigella. Although none of the carrot samples from the field were positive for E. coli, one sample was positive for Salmonella, and another was positive for F+ coliphages. From the truck, two carrot samples (8%) were positive for Salmonella, four (16%) were positive for F+ coliphages, and four (16%) were positive for E. coli. None of the carrot samples from the processing shed were positive for Salmonella. However, 2 carrot samples (8%) were positive for E. coli, and 14 carrot samples (56%) were positive for F+ coliphages. The PFGE results suggest that there were three distinct Salmonella genotypes among the carrot samples from the truck and that the Salmonella isolates identified on carrot samples from the field and truck locations were different. Microbiological screening of fresh produce such as carrots (which can be exposed to fecal contaminants in soils and water) should ensure the detection of both viral and bacterial contaminants. Overall, in this study, F+ coliphages were detected in 25% of the carrot samples, compared to E. coli (8%), Salmonella (4%), and Shigella (0%). The results suggest F+ coliphages can serve as a conservative indicator of fecally associated viruses on carrots. This suggests that in addition to E. coli screening, F+ coliphages should be included when produce such as carrots that are vulnerable to fecal contaminants are screened. Since the detection of specific enteric viral pathogens is expensive, screening for viral indicators of fecal contamination using F+ coliphages can be an economical approach to providing an additional level of assurance about the microbiological quality of fresh carrots. PMID- 12540187 TI - Resistance of bacterial isolates from poultry products to therapeutic veterinary antibiotics. AB - Bacterial isolates from poultry products were tested for their susceptibility to 10 antibiotics commonly used in the therapeutic treatment of poultry. Bacteria were isolated from fresh whole broiler carcasses or from cut-up meat samples (breast with or without skin, wings, and thighs) that were either fresh or stored at 4 or 13 degrees C (temperatures relevant to poultry-processing facilities). The Biolog system was used to identify isolates, and a broth dilution method was used to determine the antibiotic resistance properties of both these isolates and complementary cultures from the American Type Culture Collection. The antibiotics to which the most resistance was noted were penicillin G, sulfadimethoxine, and erythromycin; the antibiotic to which the least resistance was noted was enrofloxacin. Individual isolates exhibited resistances to as many as six antibiotics, with the most common resistance pattern involving the resistance of gram-negative bacteria to penicillin G, sulfadimethoxine, and erythromycin. Differences in resistance patterns were noted among 18 gram-positive and 7 gram negative bacteria, and comparisons were made between species within the same genus. The data obtained in this study provide a useful reference for the species and resistance properties of bacteria found on various raw poultry products, either fresh or stored at temperatures and for times relevant to commercial processing, storage, and distribution. The results of this study show that resistance to antibiotics used for the therapeutic treatment of poultry occurs in bacteria in the processing environment. PMID- 12540188 TI - A competitive polymerase chain reaction-based approach for the identification and semiquantification of mitochondrial DNA in differently heat-treated bovine meat and bone meal. AB - The risk of bovine spongiform encephalopathy propagation was drastically reduced after the European Union (EU) Health Authorities adopted restrictions involving a ban on animal-derived proteins in the diet of farm animals. Currently, the EU's officially recommended method for controlling meat and bone meal (MBM) in animal feed is the microscopic method, which involves the identification of bone fragments on the basis of their morphological characteristics. Recently, we demonstrated that a polymerase chain reaction (PCR)-based assay can be used for the detection of taxon-specific DNA in MBM and animal feeds. To ensure the safe rendering of animal by-products, the EU Council requires that this material be treated at 133 degrees C at 300 kPa for 20 min. Here we investigate the relationship between DNA degradation, PCR amplification, and MBM heat treatment. With a competitive PCR-based approach, we compare the amplification efficiency of bovine mitochondrial DNA target sequences of different lengths in several heat treated MBM samples. For our method, a synthetic competitive DNA is used as an internal control for both DNA extraction and PCR reaction. A correlation between an increase in treatment temperature and a reduction in the size of the target sequences suitable for amplification was observed, suggesting progressive DNA fragmentation due to the temperature. We show that short amplicons (147 bp) can be used to detect the presence of bovine mtDNA in MBM samples treated according to the current European regulations. The use of such a competitive approach to compare amplification efficiency levels of targets of different lengths might represent a useful tool for the determination of both the amount of MBM in animal feeds and its proper heat treatment. PMID- 12540189 TI - Tetrodotoxin-associated snail poisoning in Zhoushan: a 25-year retrospective analysis. AB - Outbreaks of paralytic snail poisoning have recently occurred in Asia, especially in China. The epidemiological characteristics of this disease from an outbreak in Zhoushan City, China, were recorded. Forty-two outbreaks of paralytic snail poisoning, involving 309 cases of illness, occurred from 1977 to 2001. Sixteen people (5.2%) died, 48 people (15.5%) required intubations, and 140 people (45.3%) required emergency hospital treatment as a result of these outbreaks. Outbreaks involved multiple marine snail species and occurred primarily during the summer (from June to August) on 11 islands with high population densities. Peak numbers of outbreaks and amounts of snail toxicity occurred from 1978 to 1979, from 1985 to 1987, and from 1992 to 1994. Toxicity varied depending on specimen, region, and season. The toxin involved was identified as tetrodotoxin. The data obtained in this study suggest that snails should not be eaten unless they are certified to be nontoxic. PMID- 12540190 TI - Effectiveness of commercial disinfectants for inactivating hepatitis A virus on agri-food surfaces. AB - Six commercial disinfectants were tested for their efficacy in inactivating hepatitis A virus in solution or attached to agri-food surfaces. Disinfectant I contains 10% quaternary ammonium plus 5% glutaraldehyde; disinfectant II contains 12% sodium hypochlorite; disinfectant III contains 2.9% dodecylbenzene sulfonic acid plus 16% phosphoric acid; disinfectant IV contains 10% quaternary ammonium; disinfectant V contains 2% iodide; and disinfectant VI contains 2% stabilized chlorine dioxide. Among these, disinfectants I and II were shown to be the most effective in inactivating hepatitis A virus in solution. The efficacy of these disinfectants was further tested against hepatitis A virus attached to common agri-food surfaces, including polyvinyl chlorine, high-density polyethylene, aluminum, stainless steel, and copper. Disinfectant II was shown to be the most effective, with a maximum inactivation level of about 3 log10. The inactivation efficacy was shown to be affected by the concentration of the active ingredient, the contact time between the disinfectant and the contaminated surfaces, and the incubation temperature. In general, hepatitis A virus was shown to be highly resistant to most disinfectants tested, and high concentrations of active ingredient were needed to achieve acceptable inactivation levels. PMID- 12540191 TI - Modification of an approved medium for fecal coliform detection in seawater: A-1 medium minus salicin. AB - Four hundred fifteen shellfish seawater samples from approved, conditionally approved, and restricted areas along the coastlines of Connecticut, Massachusetts, and Maine were tested in duplicate to compare results obtained with A-1 medium (AOAC official method no. 978.23) and those obtained with A-1 medium without salicin. Four laboratories used five sets of most probable number procedures to perform the analyses. No statistically significant differences between the two media were found with the t test, the F test, or the analysis of variance. PMID- 12540192 TI - Absence of nematodes in farmed Atlantic salmon (Salmo salar L.) in Norway. AB - In the present study, a total of 1,180 samples of muscle or viscera from Norwegian-farmed salmon (Salmo salar L.) were examined for the presence of nematode larvae. The samples represented all salmon-producing counties in Norway. The method applied was based on the degradation of fish soft tissue in an acidified pepsin enzyme solution. None of the samples examined in this study contained nematodes. PMID- 12540193 TI - Oyster-to-oyster variability in levels of Vibrio parahaemolyticus. AB - This study examined the variability in the levels of total and pathogenic Vibrio parahaemolyticus in individual oysters. Twenty oysters were collected on three occasions (in June, July, and September 2001) from a site near Mobile Bay, Ala. Ten of these oysters were tested immediately, and 10 were tested after 24 h of storage at 26 degrees C. Levels of total and pathogenic V. parahaemolyticus were determined by alkaline phosphatase-labeled DNA probe procedures targeting the thermolabile hemolysin and thermostable direct hemolysin genes, respectively. Similar V. parahaemolyticus levels (200 to 2,000 CFU/g) were found in nearly 90% of the oysters (for all sampling occasions) prior to storage. The log-transformed densities (means +/- standard deviations) of V. parahaemolyticus in oysters immediately after harvest were 2.90 +/- 0.91, 2.88 +/- 0.36, and 2.47 +/- 0.26 log10 CFU/g for June, July, and September, respectively. After storage for 24 h at 26 degrees C, the mean V. parahaemolyticus densities increased approximately 13- to 26-fold. Before storage, pathogenic V. parahaemolyticus was detected in 40% (10 to 20 CFU/g) of the oysters collected in June and July but was not detected in any oysters collected in September. After storage, pathogenic V. parahaemolyticus was detected in some oysters at levels of > 100 CFU/g. These data should aid in the development of sampling protocols for oyster monitoring programs and in the determination of exposure distributions associated with raw oyster consumption. PMID- 12540194 TI - Consumer food handling in the home: a review of food safety studies. AB - Epidemiological data from Europe, North America, Australia, and New Zealand indicate that a substantial proportion of foodborne disease is attributable to improper food preparation practices in consumers' homes. International concern about consumer food safety has prompted considerable research to evaluate domestic food-handling practices. The majority of consumer food safety studies in the last decade have been conducted in the United Kingdom and Northern Ireland (48%) and in the United States (42%). Surveys (questionnaires and interviews), the most frequent means of data collection, were used in 75% of the reviewed studies. Focus groups and observational studies have also been used. One consumer food safety study examined the relationship between pathogenic microbial contamination from raw chicken and observed food-handling behaviors, and the results of this study indicated extensive Campylobacter cross-contamination during food preparation sessions. Limited information about consumers' attitudes and intentions with regard to safe food-handling behaviors has been obtained, although a substantial amount of information about consumer knowledge and self reported practices is available. Observation studies suggest that substantial numbers of consumers frequently implement unsafe food-handling practices. Knowledge, attitudes, intentions, and self-reported practices did not correspond to observed behaviors, suggesting that observational studies provide a more realistic indication of the food hygiene actions actually used in domestic food preparation. An improvement in consumer food-handling behavior is likely to reduce the risk and incidence of foodborne disease. The need for the development and implementation of food safety education strategies to improve specific food safety behaviors is reviewed in this paper. PMID- 12540195 TI - Biochemistry and molecular biology of gelatinase B or matrix metalloproteinase-9 (MMP-9). AB - The matrix metalloproteinases (MMPs) form an enzyme family of which gelatinase B (MMP-9) represents the largest and most complex member. We focus here on the biochemical properties, regulation, and functions of gelatinase B. The tight regulation of gelatinase B activity is highly complex and is established at five different levels. The transcription of the gelatinase B-gene depends on various cis-elements in its gene promotor and is induced or repressed by a large variety of soluble factors, including cytokines, growth factors, and hormones and by cellular contacts acting through specific signaling pathways. The specific regulation of its secretion occurs in cells storing gelatinase B in granules. After secretion, progelatinase B must be activated through an activation network. The enzyme activity is further regulated by inhibition and by other mechanisms, such as fine-tuning and stabilization by glycosylation. The ability of gelatinase B to degrade components of the extracellular matrix and to regulate the activity of a number of soluble proteins confers an important role in various physiological and pathological processes. These include reproduction, growth, development, inflammation, and vascular and proliferative diseases. PMID- 12540196 TI - Transcription in the archaea: basal factors, regulation, and stress-gene expression. AB - A brief survey is presented of salient findings on transcription in the Archaea, focussing on stress genes of the hsp70(dnaK locus, which code for the molecular chaperones Hsp70(DnaK), Hsp40(DnaJ), and GrpE. Archaeal basal factors and some recently characterized regulators pertinent to non-stress genes are presented first to show their similarities and differences with equivalents in organisms of the other two phylogenetic domains, Bacteria and Eucarya, and to reveal clues on how these or similar factors might transcribe and regulate the archaeal stress genes. The second part of the article deals with the hsp70(dnaK)-locus genes, particularly those from Methanosarcina mazeii, because they are virtually the only ones within the methanogenic Archaea whose patterns of constitutive and stress-induced expressions have been studied. Therefore, these genes provide a standardized model system to elucidate transcription initiation and regulation at the molecular level in this phylogenetic group. Promoters, and other cis-acting sites that are, or might be, involved in stress-gene expression are described. Conformational changes of basal transcription factors after interaction with stress-gene promoters are discussed that suggest ways for generating a large diversity of initiation complexes using a few factors and DNA sites in different combinations. Likewise, the effects of stress on DNA topology and on TBP-TFB promoter complex formation and tightness are described, which might also contribute to the generation of transcription-initiation complex diversity. This diversity would be key to differential gene expression, namely, which genes are transcribed, when (basal, steady expression vs. sporadic stress-induced expression), and to what level. Future research should investigate this diversity, and the mechanism of complex formation and action at the atomic, molecular, and supramolecular levels, to elucidate the dynamics of transcription initiation in real time. PMID- 12540197 TI - World technology panel report on tissue engineering. PMID- 12540198 TI - Fiber alignment imaging during mechanical testing of soft tissues. AB - A method to image fiber alignment during mechanical testing of soft tissues was developed based on quantitative polarized light microscopy. Images were acquired after passing light through a rotating polarizer, a tissue sample, and an effective circular analyzer at multiple polarizer positions during uniaxial mechanical testing. The image set was analyzed offline using harmonic analysis to generate an alignment image, which contains the direction and strength of alignment at each image pixel. Alignment images of the entire tissue sample were generated every 3-5 s during the mechanical test allowing stress-strain behavior to be correlated with fiber alignment. Loading of fresh tissue-equivalent samples in the direction normal to the initial direction of fiber alignment revealed a spatially inhomogeneous realignment into the loading direction, with most realignment occurring near the free edges undergoing maximum lateral contraction and prior to significant load developing. Glutaraldehyde-fixed samples, in contrast, showed little realignment until yielding occurred. PMID- 12540199 TI - Effects of swelling pressure and hydraulic permeability on dynamic compressive behavior of lumbar annulus fibrosus. AB - The objective of this study was to investigate the effects of swelling pressure and hydraulic permeability on the dynamic behavior of intervertebral disk tissue in confined compression. Normal (served as a control) and trypsin-treated, axial annulus fibrosus (AF) specimens from the porcine lumbar disks were tested and their swelling strain, swelling pressure, equilibrium compressive modulus (HA), dynamic modulus, and hydraulic permeability (k) were determined at 30% and 40% swelling strain levels. The proteoglycan depletion due to trypsin treatment resulted in significantly lower values of the free swelling strain, swelling pressure, equilibrium modulus, dynamic modulus, and higher value of hydraulic permeability for trypsin-treated group, comparing to those for the control group. At the 30% swelling strain level, the equilibrium moduli were 51.84 +/- 14.53kPa (n = 8) for the control group and 15.11 +/- 5.67 kPa (n = 8) for the trypsin treated group; and the hydraulic permeabilities were 4.50E- 15 +/- 1.60E- 15 m4/Ns and 8.43E-15 +/- 4.29E- 15 m4/Ns for control and trypsin-treated groups, respectively. No statistically significant difference in wet tissue density or dry tissue density was found between control and trypsin-treated groups. There was a significant correlation between swelling pressure and compressive (aggregate) modulus (R2 = 0.824, m = 22). The decrease in measured dynamic modulus for trypsin-treated group was attributed to the reduced swelling pressure (or modulus HA) and increased hydraulic permeability (k) due to PG depletion. PMID- 12540200 TI - A device for imposing cyclic strain to cells growing on implant alloys. AB - The objective of this study was to develop a unique device for applying well characterized cyclic, strain to cells growing on implant alloys. The device is based on the four-point bending principle and utilizes an electric motor, belt, and cam system to cyclically deflect a commercially pure titanium plate with cell culture wells in the middle of the plate. Analyses demonstrated that 182 +/- 3, 366 +/- 9, and 984 +/- 7 microstrain may be cyclically generated in culture areas from 0.5 to 10 Hz for up to 72 h. UMR-106 osteoblast-like cells growing on the titanium plate were subjected to these strain magnitudes at 1.5 Hz for periods of 4 or 24 h. Cells were checked for viability, total protein as a general indicator of cell number, and alkaline phosphatase activity (ALP) as an indicator of bone cell func-tion. Cells strained at 984 +/- 7 microstrain exhibited 21%-24% more protein but 45%-49% less ALP activity than cells strained at 182 +/- 2 or 366 +/- 9 microstrain. Decreased ALP activity may indicate impaired mineralization. Results indicate the device is suitable for applying known, cyclic strain to cells growing on implant alloys and evaluating cellular responses to strain while growing on implant alloys. PMID- 12540201 TI - The effect of pulsed ultrasound on mandibular distraction. AB - This study evaluated the effect of pulsed ultrasound on tissue repair and bone growth during mandibular osteodistraction. Twenty-one rabbits were divided into three groups of 7. The distraction started 72 h after surgically severing both sides of the mandible and proceeded at a rate of 1.5 mm/12 h for 5 days. Group I received pulsed ultrasound (nominally 200 micros pulse of 1.5 MHz at a 1.1 kHz pulse repetition frequency, 30 mW/cm2) for 20 min on both sides of the mandible every other day (alternating sides). Group 2 received the same pulsed ultrasound treatment on one side of the mandible every day for 20 min. Group 3 did not receive any ultrasound treatment. Bone formation at the distraction site was assessed by photodensitometry on head radiographs, a vibratory coherence test across the distraction site, a postmortem three-point bending mechanical stiffness test, and a postmortem histological examination. Statistical analyses performed using analysis of variance revealed that pulsed ultrasound enhanced bone formation at the distraction site with a high level of significance when assessed by the increase in new bone photodensity (p = 0.001), vibratory coherence (p = 0.001), mechanical stiffness (p = 0.003), and qualitative histological studies, especially when the pulsed ultrasound treatment was directly applied daily. PMID- 12540202 TI - Inhibition of shear-stress-induced platelet aggregation and phosphotyrosine signaling by GPIIb-IIIa antagonists. AB - Shear-stress-mediated platelet thrombus formation has been implicated in the pathophysiology of cardiovascular diseases such as acute myocardial infarction and unstable angina. Although previous studies have established that fluid shear forces cause platelet aggregation, a direct comparison of GPIIb-IIIa antagonists used in the treatment of acute coronary syndromes on shear-induced platelet activation has not been reported. Therefore, the objective of the present study was to characterize the effects of the platelet antagonists abciximab, eptifibatide, and tirofiban on shear-mediated platelet activation and aggregation using flow cytometric and Western blotting techniques. Flow cytometric analyses indicated that all three platelet antagonists, when used at concentrations that saturated all GPIIb-IIIa receptors, significantly inhibited platelet aggregate formation and expression of the platelet activation marker p-selectin. None of the antagonists caused increased expression of GPIbalpha or GPIIb-IIIa on the platelet surface compared to untreated controls. Additionally, Western blotting demonstrated that a 72 kDa protein tentatively identified as Syk became phosphorylated in response to shear stress and that its phosphorylation was inhibited by each antagonist. The findings of this study indicate that abciximab, eptifibatide, and tirofiban, though possessing distinct biochemical and pharmacological properties, effectively and equivalently inhibit platelet aggregation, p-selectin expression, and intracellular tyrosine phosphorylation events induced by fluid shear stress. PMID- 12540203 TI - Experimental studies of membrane tethers formed from human neutrophils. AB - Membrane tethers (thin, cylindrical pieces of membrane) have been implicated in the rolling of neutrophils along the endothelium. In our studies, these tethers were formed from passive, stimulated (0.1 microM fMLP), and osmotically swollen (170-180 mOsm) human neutrophils; as well as neutrophils treated with 0.3 microM latrunculin A to disrupt the cytoskeleton. This tether formation was accomplished by micropipette suction of latex beads coated with antibodies to proteins on the neutrophil membrane surface. From plots of force versus velocity for the tether formation process, we calculated adhesion energies per unit area of the lipid membrane to the cytoskeleton and the viscous resistance (effective viscosity) that occurs during the formation of these tethers at finite velocity. Most of the properties of the neutrophil were altered once it had been treated as described above. We were also able to show mechanical reversibility of membrane tethers, as well as an unexpected formation rate at "high" tether forces. Since membrane tethers have been implicated in the rolling of neutrophils, then the changes in tether formation may ultimately alter how these cells roll. PMID- 12540204 TI - Surface strains in the anterior leaflet of the functioning mitral valve. AB - Abstract-The mitral valve (MV) is a complex anatomical structure whose function involves a delicate force balance and synchronized function of each of its components. Elucidation of the role of each component and their interactions is critical to improving our understanding of MV function, and to form the basis for rational surgical repair. In the present study, we present the first known detailed study of the surface strains in the anterior leaflet in the functioning MV. The three-dimensional spatial positions of markers placed in the central region of the MV anterior leaflet in a left ventricle-simulating flow loop over the cardiac cycle were determined. The resulting two-dimensional in-surface strain tensor was computed from the marker positions using a C0 Lagrangian quadratic finite element. Results demonstrated that during valve closure the anterior leaflet experienced large, anisotropic strains with peak stretch rates of 500%-1,000%/s. This rapid stretching was followed by a plateau phase characterized by relatively constant strain state. We hypothesized that the presence of this plateau phase was a result of full straightening of the leaflet collagen fibers upon valve closure. This hypothesis suggests that the MV collagen fibers are designed to allow leaflet coaptation followed by a dramatic increase in stiffness to prevent further leaflet deformation, which would lead to valvular regurgitation. These studies represent a first step in improving our understanding of normal MV function and to help establish the principles for repair and replacement. PMID- 12540206 TI - Torso coupling techniques for the forward problem of electrocardiography. AB - The calculation of body surface potentials from a known cardiac source is traditionally formulated in terms of a two step process. The first step involves the generation of some form of equivalent cardiac source (typically dipole based) at a resolution significantly lower than that of a continuum cell. The second then places that source into a volume conductor within which the potential fields are calculated. This approach does not properly capture the feedback between the torso and the extracellular potential field. Presented here are the details of two new methods which calculate continuous potential fields throughout the torso that are the direct result of cardiac cellular electrical activity. These new methods are termed the Boundary Iteration Method and the Direct Assembly Method. While these two methods return essentially identical answers, there is a definite tradeoff between computational speed and memory overhead with the direct assembly method proving faster but requiring significantly more memory. Examples are given that demonstrate the convergence and accuracy of these methods in an idealized coupled torso system. These techniques are then applied to an anatomically based model of a slice through a human male torso. PMID- 12540205 TI - Length-normalized pulse photoplethysmography: a noninvasive method to measure blood hemoglobin. AB - To determine whether blood hemoglobin concentration ([Hb]) could be measured noninvasively as the ratio of pulsatile changes in attenuation (absorbance plus scatter) of light (D) across a body part to changes in light path length (l), we measured transmission of near-infrared light (905 +/- 10 nm) through a finger, using a modified pulse oximeter, and simultaneously monitored fingertip diameter, using a sonomicrometer. In 25 subjects with [Hb] ranging from 3.1 to 18.2 gm/dl, and with normal oxygenation, average D/l ratio over 30-60 s correlated strongly with [Hb] measured by Coulter counter (r = 0.84, p << 0.001), though with considerable scatter, with absolute value of differences averaging 17% of the mean. Using 12 gm/dl and 0.75 mm(-1) as the lower limits of normal for [Hb] and D/l, respectively, two of nine normals had low (D/l) (78% specificity), and only one of 16 anemic subjects had borderline normal (D/l) (94%-100% sensitivity). The positive predictive value of a low (D/l) was 88% and the negative predictive value was 87.5%. With further development, this technique may reduce the need for phlebotomy, thereby reducing risks and costs and improving the experience of being a patient. PMID- 12540207 TI - A three-dimensional numerical fluid dynamic model of antigen-antibody surface adsorption on piezoelectric immunosensors. AB - A piezoelectric crystal is a unit that changes its frequency in parallel with a change in its mass. This characteristic is exploited in designing flow cell-based immunosensors for detecting the concentration of antibodies in liquid samples. In the present study, computational fluid dynamic techniques are used to optimize the antigen-antibody binding process on an electrode surface placed on the base of a conical flow cell. The geometry optimization of the flow cell was determined to minimize the test time. This time is needed for the electrode to be saturated by the antibody, a process that requires the maximization of the adsorption rate and be accomplished by increasing the shear rate in the vicinity of the electrode. To validate the numerical model and to determine its parameters, experiments were carried out using an identical flow cell. In the experiments, the system did not reach saturation within an acceptable time frame, therefore, the model parameters were determined based on the unsaturated state. The experimental results confirmed the applicability of numerical simulations in predicting the effect of changing the inlet section area of the flow cells, proving the computational model to be very valuable in designing immunosensors based on flow cells. PMID- 12540208 TI - An in vitro evaluation of the antibacterial properties of barriers used in guided tissue regeneration. AB - The inhibition of growth of Prevotella nigrescens and Enterococcus faecalis by three barriers membrane, (a) polylactic acid (Atrisorb), (b) laminar bone, and (c) calcium sulfate, was compared using an agar diffusion test. The first group of 20 plates were inoculated with P. nigrescens in a prereduced anaerobically sterilized brain-heart infusion broth, while the second group of 20 plates were inoculated with E. faecalis in a brain-heart infusion broth. On each plate, a 5.0 mm section of each barrier was placed such that every plate had all three barriers. The plates in the P. nigrescens group were incubated in an anaerobic chamber for 7 days. The plates in the E. faecalis group were incubated at 37 degrees C for 2 days. No zones of inhibition were observed with laminar bone or calcium sulfate. Zones of inhibition were observed with the Atrisorb barrier membrane in both groups. The zone of inhibition of P. nigrescens was significantly greater than the zone of inhibition of E. faecalis. The hydrophobicity of Atrisorb and its N-methyl-2-pyrolidine component were suggested as the sources of its antibacterial activity. PMID- 12540209 TI - pH changes in root dentin after intracanal placement of improved calcium hydroxide containing gutta-percha points. AB - The in vitro pH changes in root dentin over a period of 2 weeks was investigated in 48 extracted bicuspids after intracanal placement of either Roeko Calcium Hydroxide Plus Points, aqueous calcium hydroxide paste, or gutta-percha points (control group) after root canal preparation. Microelectrodes were placed in outer and inner root dentin at cervical, middle, and apical thirds of the root to measure the pH at 1 h, 2 h, 3 h, 1 day, 3 days, 7 days, and 14 days. Roeko Calcium Hydroxide Plus Points reached a peak inner dentine pH of 11.67 and a peak outer dentine pH of 10.82 at 3 h. In addition, Roeko Calcium Hydroxide Plus Points maintained an outer dentine pH above 9.5 for approximately 2 days, whereas the aqueous calcium hydroxide paste did not reach this pH. However, the alkalinity in dentin with Roeko Calcium Hydroxide Plus Points was maintained for 7 days only, whereas aqueous calcium hydroxide paste maintained an alkaline environment throughout the 2-week period. PMID- 12540210 TI - Proliferative ability and alkaline phosphatase activity with in vivo cellular aging in human pulp cells. AB - Little is known about the effect of aging on characteristic functions of pulp cells. When damaged pulp is recovered and mineralized tissue is formed to protect remaining pulp tissue, the general responses of pulp tissue after adequate stimuli (pulp cell proliferation and activation of alkaline phosphatase [ALPase]) are thought to be essential. In this study, we compared proliferative ability and ALPase activity between cultures of human pulp (HP) cells obtained from young and aged donors. The in vitro proliferative lifespan of HP cells from young donors was longer than HP cells from aged donors. Growth rates and ALPase activity of HP cells decreased with increasing donor age. These findings suggest that impaired repair of pulp and dentin in aged patients is partly due to a decrease in the proliferative ability and ALPase activity in aged pulp cells. PMID- 12540211 TI - Bacteriological evaluation of passive ultrasonic activation. AB - The purpose of this study was to evaluate the influence of passive ultrasonic activation on root canal disinfection. Sixty human teeth (group A: upper incisors, group B: upper canines, and group C: distobuccal root of first upper molars) were selected and sterilized in an autoclave. A standardized inoculum was placed into the canals, and they were incubated for 72 h at 37 degrees C. Then, they were divided into subgroup 1, which received sterile saline (SS) as an irrigant, and subgroup 2, which received sterile saline with passive ultrasonic activation (SU). The endodontic treatment was performed with a crown-down technique. Bacteriological identification of surviving colonies was carried out. Surviving colonies were higher when ultrasonics was not used (group A: SS: x 32.13, SU: x 13.53; group B: SS: x 53.70, SU: x 44.60; group C: SS: x 39.16, SU: x 29.40). The homogeneity proportion tests to compare the results of both subgroups showed that the surviving proportions were higher (p = 0.01) when the ultrasonic activation was not used. PMID- 12540212 TI - Comparative analysis of torsional and bending stresses in two mathematical models of nickel-titanium rotary instruments: ProTaper versus ProFile. AB - During root canal instrumentation, nickel-titanium rotary instruments are subjected to continual stresses inside the canal due to its anatomy and the hardness of the dentin they must cut. They must therefore be both stress resistant and elastic. This study aimed to compare the mechanical behavior of two nickel-titanium rotary instruments (ProTaper and ProFile) by applying the finite element analysis method to produce a numerical evaluation. The nonlinear mechanical behavior of the alloy was taken into account during the study. The distribution of stresses due to torsional and bending moments was compared in the two experimental models. The ProFile model was found to be more elastic than the ProTaper model. Under equal loads, the ProTaper model showed lower and better distributed stresses than the ProFile model. PMID- 12540213 TI - Properties of a new injectable type of root canal filling resin with adhesiveness to dentin. AB - The properties of a new, injectable type of root canal wall, adhesive filling resin, which we developed, were studied for its physical properties, adhesiveness to dentin and the root canal, and sealing ability. The new resin consisted of a powder composed of a specially selected poly(methyl methacrylate) and barium sulfate radiopacifier and liquid composed of methyl methacrylate monomer and tributylborane catalyst. A mixture of both components had a consistency suitable for injection. The physical properties, including radiopacity, were evaluated according to the ISO standard. The adhesiveness to dentin and the root canal was investigated by bond strength testing or by scanning electron microscopy. The apical sealing ability of the new resin was compared by a dye penetration test with that of gutta-percha/sealer. The findings of this study indicated that the new filling resin had physical properties satisfying all the ISO requirements for dental root canal sealing materials: a tensile bond strength to dentin of 7.3 MPa, a significantly better sealing ability compared with gutta-percha/sealer, the capability of forming resin tags in dentinal tubules, and removability. PMID- 12540214 TI - Bacterial concentrations determine the ability to implant in the root canal system and translocate to lymph nodes in germ-free mice. AB - The purpose of this study was to evaluate the influence of the infecting dose on bacterial colonization in root canal systems (RCS) and translocation to the submandibular lymph node (SML) of gnotobiotic (germ-free) mice. RCS were inoculated with 10(2) and 10(1) colony-forming units (CFU) of Enterococcus faecalis (ATCC 4083) or 10(5), 10(4), 10(3), and 10(2) CFU of Prevotella nigrescens (ATCC 33563). E. faecalis implanted in 83.3% of the cases, for both inocula. Translocation was detected in 58.3% of lymph nodes for the 10(2) CFU inoculum and in 33.3% of lymph nodes for the 10(1) CFU inoculum. P. nigrescens implanted in 25%, 16.7%, 8.3%, and 0%, for the 10(5), 10(4), 10(3), and 10(2) CFU inocula, respectively, and did not translocate at any of the concentrations used. These results indicate that E. faecalis was able to implant in the RCS and translocate to the SML, whereas P. nigrescens implanted in lower percentages in the RCS and did not translocate to the SML. PMID- 12540215 TI - The inhibitory effect of alendronate and taurine on osteoclast differentiation mediated by Porphyromonas gingivalis sonicates in vitro. AB - The objective of this study was to investigate the ability of alendronate and taurine in inhibiting in vitro osteoclast differentiation induced by bacteria. Whole cell sonicates of Porphyromonas gingivalis were used as an osteoclast stimulating factor in a mouse coculture system and differentiated osteoclasts were confirmed by tartrate-resistant acid phosphatase (TRAP) staining. Alendronate at the concentrations of 10(-7) M, 10(-6) M, and 10(-5) M and taurine at the concentrations of 4 mM, 8 mM, and 12 mM were used. The cytotoxic effects of alendronate and taurine were examined using methylthiazole-tetrazolium bromide (MTT) assay. The amounts of interleukin-6 (IL-6) in culture supernatants were also measured using ELISA. The sonicates of P. gingivalis at the concentration of 0.01-0.1 microg/ml significantly stimulated the formation of osteoclasts (p < 0.05). Alendronate (10(-5) M) and taurine (12 mM) significantly suppressed the sonicate-stimulated osteoclast formation. In MTT assay, no cytotoxic effects were evident in all concentrations of alendronate and taurine. Alendronate and taurine did not affect the amount of IL-6 induced by P. gingivalis sonicates. These data indicate that alendronate and taurine have inhibitory effects on bacteria stimulated osteoclast formation in vitro and that this inhibitory mechanism is not related to the blocking of IL-6 production. PMID- 12540217 TI - In vitro cytotoxicity of medicated and nonmedicated gutta-percha points in cultures of gingival fibroblasts. AB - This investigation was designed to test the cellular toxicity of two medicated (Roeko activ point and Roeko Calcium Hydroxide) and four nonmedicated brands of gutta-percha (GP) points (Antaeos, DeTrey White, Roeko color, and Roeko Top color). The test points were transferred into a culture medium including the GP point material with a concentration of 6 mg/ml, and eluates were obtained after 72 h. Five milliliters of each eluate were pipetted onto fibroblast cultures, incubated, and subsequently stained. Mitotic rates, cell densities, and the distribution of normal cells, pathologically altered and dead cells were determined and correlated with control cell cultures. Roeko activ point (containing chlorhexidine) resulted in the highest number of dead cells. The difference was statistically significant in comparison with all other materials. Concerning all parameters mentioned, the cytotoxicity of the points containing calcium hydroxide (Roeko Calcium Hydroxide) was not significantly different from all other points tested, with the exception of those containing chlorhexidine. All tested gutta-percha materials caused cytotoxic reactions in varying extents. Taking into consideration the limitations of an in vitro experiment, points containing calcium hydroxide and nonmedicated points seem to be the most recommendable products for clinical use. PMID- 12540216 TI - The effect of irrigation with doxycycline or citric acid on leakage and osseous wound healing. AB - This study investigated the effect of doxycycline irrigation on wound healing and the apical seal of three filling materials. Part 1: 220 extracted teeth received root canal therapy followed by root resection (2 mm) and ultrasonic root end preparations (3 mm). Groups of 20 were irrigated with saline, citric acid, or doxycycline and filled with amalgam, Super EBA, or MTA. Leakage was measured (mm) after decalcification and clearing. Part 2: two defects were made on each side of the mandible of 10 New Zealand rabbits. On each side, one defect was irrigated with saline and one with either citric acid or doxycycline. The animals were killed in groups of five at 9 and 18 days. Sections of each defect were stained with hematoxylin and eosin for evaluation of healing and bone fill. Super EBA and MTA leaked significantly less than amalgam, regardless of irrigant. Leakage after irrigation with doxycycline compared to citric acid or saline was not significantly different for Super EBA or MTA but was lower for amalgam. There was no significant difference in healing or bone fill among irrigants at 9 or 18 days. PMID- 12540218 TI - Differential induction of apoptosis by capping agents during pulp wound healing. AB - Effects of capping agents on two waves of apoptosis during pulp wound healing were examined. After cavity preparation of rat molars, cavities were filled with calcium hydroxide, zing oxide eugenol cement, or 4-META/MMA-TBB resin (4MMT) and some were unfilled. One hour or 1 day after filling, we examined the distribution and the number of apoptotic cells by terminal deoxynucleotidyl transferase mediated labeling (TUNEL) assay. One hour after filling, there were no differences in distribution patterns or the number of apoptotic odontoblasts among the four groups. One day after filling, we found differences in distribution patterns and the number of apoptotic pulp cells among the four groups. Especially in the 4MMT group, the distribution pattern of apoptotic cells was more broadly spread, and the number of apoptotic cells was significantly larger than those of other groups. These results suggest that capping agents may have an effect on pulp apoptosis and that 4MMT may actively induce apoptosis during pulp wound healing. PMID- 12540219 TI - Antibiotic susceptibility of bacteria associated with endodontic abscesses. AB - Antibiotics to treat endodontic infections are routinely prescribed based on previously published susceptibility tests. There is increased concern that bacteria have increased resistance to the currently recommended antibiotics. The purpose of this investigation was to perform antibiotic susceptibility tests on a panel of bacteria recently isolated from endodontic infections. The bacteria in this study were aseptically aspirated with a needle from endodontic abscesses, cultivated, and identified at the species level. Each of the 98 species of bacteria was tested for antibiotic susceptibility to a panel of six antibiotics using the Etest. The antibiotics were penicillin V, amoxicillin, amoxicillin + clavulanic acid, clindamycin, metronidazole, and clarithromycin. The percentages of susceptibility for the 98 species were penicillin V: 83/98 (85%), amoxicillin: 89/98 (91%), amoxicillin + clavulanic acid: 98/98 (100%), clindamycin: 94/98 (96%), and metronidazole: 44/98 (45%). Metronidazole had the greatest amount of bacterial resistance; however, if it is used in combination with penicillin V or amoxicillin, susceptibility of the combination with penicillin V or amoxicillin increased to 93% and 99%, respectively. Clarithromycin seems to have efficacy, but it is still considered an antibiotic under investigation because the minimum inhibitory concentration has not been established. PMID- 12540220 TI - Interleukin-10 receptor expression in human dental pulp cells in response to lipopolysaccharide from Prevotella intermedia. AB - Interleukin-10 receptor (IL-10R) expression in human, dental pulp, fibroblast cultures was investigated by using the reverse transcriptase-polymerase chain reaction (RT-PCR) technique. After exposure to lipopolysaccharide (LPS) from Prevotella intermedia, the IL-10R mRNA levels increased after 4 h, peaked at 7 h, and dropped back to the unstimulated level at 24 h. Maximal production of the IL 10R protein in dental pulp fibroblast cultures was detected by Western blot analysis after 12 h of LPS stimulation. In contrast, the human skin fibroblast (SF-MA) and human monocyte (U937) cell lines expressed IL-10R mRNA. Anti-CD14 antibodies inhibited P. intermedia LPS-induced IL-10R mRNA expression. These results indicate that P. intermedia LPS induces IL-10R gene expression in human, dental pulp fibroblasts in vitro. PMID- 12540221 TI - Antimicrobial activity of Ca(OH)2 dental cements: an in vitro study. AB - The effect of five commercial calcium hydroxide root canal medications (XR-SPAD, Calcicur, Hy-cal, Root-cal, Hypo-cal) and two calcium hydroxide mixtures (with water or glycerin) on the growth of (a) three anaerobic bacteria associated with endodontic infections; and (b) saliva bacterial strains was determined using the agar diffusion inhibitory test. Statistically significant zones of bacterial growth inhibition for all the bacteria tested were observed. Calcium hydroxide mixed with glycerin showed the largest zones of bacterial inhibition, whereas Root-cal was the least active. These results underline the necessity to evaluate the antibacterial potential of endodontic medications against the different bacterial strains commonly isolated from infected root canals. PMID- 12540222 TI - A comparison of torque required to fracture three different nickel-titanium rotary instruments around curves of the same angle but of different radius when bound at the tip. AB - Instrument fracture is an unfortunate but possible sequela of instrumentation of canals, especially when the instrument is bound at the tip. The purpose of this study was to compare the torque required to fracture three file sizes of three different rotary file types around two simulated canal curvatures, gradual or acute, when the tip of the working end of the file was bound. Profile Series 29 0.04 and 0.06 taper and Profile 0.06 ISO rotary files were placed passively into simulated canal curvatures of the same angle but of different radii. The file tips were bound 2 mm from the working end and a measurable torque was applied until fracture. ANOVA with Tamhane post-hoc comparison showed that the 0.06 Series 29 did not differ from the ISO 0.06 taper or the 0.04 Series 29 but there was statistical difference (p < 0.01) showing that the 0.04 Series 29 broke with less force than did the 0.06 ISO files. Statistical tests (p < 0.01) also showed smaller files failed with less torque, as did files in more acute canal curvatures. PMID- 12540223 TI - Working length determination in palatal roots of maxillary molars. AB - The purpose of this study was to determine if a buccal curvature in the palatal roots of maxillary molars affected the clinician's ability to accurately determine working length. Twenty-seven extracted, human maxillary molars were sorted by palatal root curvatures as J- and C-type, and the angle of curvature was determined. Straight-line access was made and a #20 file was placed into the canal until the tip was visible at the apical foramen then withdrawn. The file, tooth, and calibration wire were radiographed on one image using the RadioVisioGraphy system. Actual (file) and radiographic (tooth) lengths were determined using the RadioVisioGraphy ruler. Radiographic length appeared shorter on average than the actual length. Canal curvatures larger than 25 degrees had differences greater than 0.5 mm. This represents a statistically significant difference between the actual and radiographic lengths as the degree of curvature increases. There was no significant difference between the J- and C- types. PMID- 12540224 TI - Effectiveness of prophylactic use of rofecoxib in comparison with ibuprofen on postendodontic pain. AB - The purpose of this study was to determine if prophylactic rofecoxib would significantly reduce postendodontic pain, when compared with ibuprofen or placebo. An additional objective was to establish if any relationship exists between periapical diagnosis and the need for additional medication after completion of pulpectomy. A total of 45 patients consented to a double-blind, single-dose oral administration of 50 mg of rofecoxib, 600 mg of ibuprofen, or a placebo before conventional root canal therapy. The root canal treatment was performed in two appointments. Patient-reported visual analog scale ratings of pain intensity were conducted upon initial clinical presentation and at 4, 8, 12, 24, 48, and 72 h after completion of pulpectomy. Results showed that at the 4- and 8-h periods, both rofecoxib and ibuprofen provided significantly better pain relief than placebo. At the 12- and 24-h periods, rofecoxib demonstrated significantly better pain relief than both ibuprofen and placebo. Patients with a periapical diagnosis of acute apical periodontitis showed a significantly increased need for additional medication after completion of pulpectomy compared with all other periapical diagnoses. PMID- 12540225 TI - Effect of endodontic instrument taper on post retention. AB - The purpose of this study was to evaluate the effect of various tapers of canal preparations on the retention of posts. Ninety human central or lateral mandibular incisors were prepared by removing the clinical crown at the CEJ and instrumenting the canals with instruments of varying tapers. The groups comprising 10 teeth each were divided as follows: Taper 0.02 Ni-Ti hand files (control), Taper 0.04 ProFile Series 29, Taper 0.06 ProFile Series 29, Taper 0.08 ProFile GT Rotary, Taper 0.10 ProFile GT Rotary, Taper 0.12 ProFile GT Rotary, Taper 0.08* ProFile GT Rotary 0.08 taper with #50/.12 taper in coronal third, Taper 0.10* ProFile GT Rotary 0.10 taper with #50/.12 taper in coronal third, Taper 0.12* ProFile GT Rotary 0.12 taper with #50/.12 taper in coronal third. Canals were filled with gutta-percha and AH 26 sealer using lateral condensation. Gutta-percha was removed with a heated 5/7 plugger until a level of 5 mm of gutta percha was left in the apical segment of the root. Post channel preparation was made to the appropriate size with minimal change in canal shape with the Premier IntegraPost drill, and the corresponding IntegraPost was cemented with 3M Vitremer cement. The roots were fixed in copper rings with mounting plaster and mounted on an Instron TT machine. The posts were subjected to gradually increasing vertical tensile force until dislodgment of the post occurred. Force (lb) required to dislodge the post was obtained for each tooth, and the average force of the groups was compared using a one-way ANOVA with Tukey's multiple comparisons. The highest resistance to dislodgment was obtained with Taper 0.04 (79.10 +/- 10.73), whereas the lowest was obtained by the Taper 0.02 (18.70 +/- 13.44). No statistical difference was found between six of the groups: Taper 0.08*, Taper 0.10, Taper 0.12*, Taper 0.10*, Taper 0.12, and Taper 0.06. Taper 0.08 was statistically inferior to Taper 0.04 but not Taper 0.02. The 0.04 taper seems to be the best taper for maximal resistance to post dislodgment. PMID- 12540226 TI - The effect of endodontic solutions on resorcinol-formalin paste. AB - This study determined if any of four endodontic solutions would have a softening effect on resorcinol-formalin paste and if there were any differences in the solvent action between these solutions. Resorcinol, formalin, and zinc oxide were mixed and allowed to set for 30 days. The solutions tested were 0.9% sodium chloride, 5.25% sodium hypochlorite, chloroform, and Endosolv R. Seven samples per solution were tested and an additional seven samples served as controls. Each sample was saturated with one of the solutions and the depth of penetration was tested at 1, 2, 5, 10, and 20 min using a dial strain gauge. At 2 min, sodium hypochlorite and sodium chloride had significantly greater penetration than the other groups (p < 0.0010). Sodium hypochlorite was superior to all other groups after 5 min. This study showed that both sodium chloride and sodium hypochlorite exhibited a significant softening effect within 2 min. PMID- 12540227 TI - Paradental cyst mimicking a radicular cyst on the adjacent tooth: case report and review of terminology. AB - A distinctive form of paradental cyst can occur on the buccal and apical aspects of erupted mandibular molars. This cyst has peculiar clinical and radiographic features, although the microscopic findings are the same as those of odontogenic inflammatory cysts. Diagnostic and therapeutic problems can occur when this lesion is misinterpreted as a radicular cyst. The purpose of this paper is to present an additional case of a paradental cyst in the buccal and mesial aspects of a mandibular second molar involving the apical area of a mandibular first molar. The difficulty of diagnosis, treatment, and controversies regarding terminology are also discussed. PMID- 12540228 TI - Retrograde treatment in an extensive implant-tooth-supported reconstruction. AB - Diagnosis and treatment of chronic apical periodontitis associated with an abutment tooth in an extensive tooth-implant-supported fixed prosthesis is reported. Careful, complete interpretation of clinical data was imperative because the periapical radiolucency presented around the apex of a vital tooth. A retrograde endodontic approach was used to prevent an adverse outcome to the complex rehabilitative treatment that had been performed over many years. PMID- 12540229 TI - Control your destiny. PMID- 12540230 TI - 5-HT4 receptor ligands: applications and new prospects. PMID- 12540231 TI - Phenethyl amides as novel noncovalent inhibitors of hepatitis C virus NS3/4A protease: discovery, initial SAR, and molecular modeling. AB - The discovery of novel, reversible and competitive tripeptide inhibitors of the Hepatitis C virus NS3/4A serine protease is described. These inhibitors are characterized by the presence of a C-terminal phenethyl amide group, which extends into the prime side of the enzyme. Initial SAR together with molecular modeling and data from site-directed mutagenesis suggest an interaction of the phenethyl amide group with Lys-136. PMID- 12540232 TI - Hybrid-designed inhibitors of p38 MAP kinase utilizing N-arylpyridazinones. AB - Imidazo[1,2-a]pyridyl N-arylpyridazinones were hybridized from the classic pyridinylimidazoles and the more recent dual hydrogen bond acceptors, resulting in a new structural class of selective p38 MAP kinase inhibitors. PMID- 12540233 TI - 3'-Aminoadenosine-5'-uronamides: discovery of the first highly selective agonist at the human adenosine A3 receptor. AB - Selective adenosine A(3) agonists have potential utility for the prevention of perioperative myocardial ischemic injury. Herein, we report on the discovery and synthesis of compound 7. This amino nucleoside agonist possesses unprecedented levels of selectivity for the human adenosine A(3) receptor. PMID- 12540234 TI - Ketones as building blocks for dynamic combinatorial libraries: highly active neuraminidase inhibitors generated via selection pressure of the biological target. AB - New and potent inhibitors of neuraminidase, a key enzyme in the influenza virus activity, have been discovered in dynamic combinatorial libraries based on ketones and amines as building blocks. Selective synthesis of a number of inhibitors among multiple theoretically possible combinations of building blocks is driven by the presence of the target enzyme. PMID- 12540235 TI - Human factor Xa bound amidine inhibitor conformation by double rotational-echo double resonance nuclear magnetic resonance and molecular dynamics simulations. AB - Double rotational-echo double resonance (double REDOR) NMR was used to investigate the conformation of a (13)C-, (15)N-, and (19)F-labeled inhibitor (Berlex Biosciences compound no. ZK-806299) bound to human factor Xa. Conformationally dependent carbon-fluorine dipolar couplings were measured by (13)C[(19)F] REDOR. Natural abundance carbon signals in the full-echo spectra were removed by (13)C[(15)N] REDOR. Major and minor binding modes were suggested by the NMR data, but only the former had adequate signal to noise for distance determinations. Molecular dynamics simulations restrained by double-REDOR determined intramolecular (13)C-(19)F distances revealed two models for the dominant binding mode that are consistent with the NMR data. We conclude that ZK 806299 binds similarly to both FXa. Moreover, it appears to bind to FXa in a fashion previously demonstrated for ZK-807834, a more selective FXa inhibitor. PMID- 12540237 TI - Topomer CoMFA: a design methodology for rapid lead optimization. AB - To provide an objective QSAR methodology that might accelerate lead optimization, the CoMFA and topomer technologies have been merged, with surprisingly good results. A series of input structures are each broken into two or more fragments at central acyclic single bonds, while removing any core fragment structurally common to the entire series. Standard topomer 3D models are automatically constructed for each fragment, and a set of steric and electrostatic fields ("CoMFA column") is generated for each set of topomers. Application of "topomer CoMFA" to 15 3D-QSAR analyses taken from the literature (847 structures) were all successful, with an average q(2) of 0.520 (literature average q(2) = 0.636) and an average standard deviation of true prediction (SDEP) of 0.688 (literature average SDEP = 0.553) for 133 structures. Topomer CoMFA results are particularly promising as queries into virtual libraries already composed of topomer structures, to directly seek structures having increased potency. Accordingly, in 13 of the 15 such "topomer CoMFA searches" attempted, combinations of commercially offered fragments were retrieved that were predicted to be more potent than any structure described in the original publication (average predicted potency increase = 20 x), showing in principle how optimization could occur. PMID- 12540236 TI - New amide-bearing benzolactam-based protein kinase C modulators induce enhanced secretion of the amyloid precursor protein metabolite sAPPalpha. AB - Protein kinase C (PKC) is known to participate in the processing of the amyloid precursor protein (APP). Abnormal processing of APP through the action of the beta- and gamma-secretases leads to the production of the 39-43 amino acid Abeta fragment, which is neurotoxic and which is believed to play an important role in the etiology of Alzheimer's disease. PKC activation enhances alpha-secretase activity, which results in a decrease of the amyloidogenic products of beta secretase. In this article, we describe the synthesis of 10 new benzolactam V8 based PKC activators having side chains of varied saturation and lipophilicity linked to the aromatic ring through an amide group. The K(i) values measured for the inhibition of phorbol ester binding to PKCalpha are in the nanomolar range and show some correlation with their lipophilicity. Compounds 5g and 5h show the best binding affinity among the 10 benzolactams that were synthesized. By use of a cell line derived from an AD patient, significant enhancement of sAPPalpha secretion was achieved at 1 microM concentration for most of the compounds studied and at 0.1 microM for compounds 5e and 5f. At 1 microM the enhancement of sAPPalpha secretion for compounds 5c-h is higher than that observed for the control compound 8-(1-decynyl)benzolactam (BL). Of interest is the absence of activity found for the highly lipophilic ligand 5i, which has a K(i) of 11 nM. On the other hand, its saturated counterpart 5j, which possesses a comparable K(i) and ClogP, retains activity in the secretase assay. In the hyperplasia studies, 5f showed a modest response at 100 microg and 5e at 300 microg, suggesting that 5f was approximately 30-fold less potent than the PKC activator mezerein and 100 fold less potent than TPA. 5e was approximately 3-fold less active than 5f. On the basis of the effect of unsaturation for other potent PKC ligands, we would predict that 5e would retain biological activity in most assays but would show a marked loss of tumor-promoting activity. Compound 5e thus becomes a viable candidate compound in the search for Alzheimer's therapeutics capable of modulating amyloid processing. PMID- 12540238 TI - Synthesis, anti-HIV activity, and molecular mechanism of drug resistance of L 2',3'-didehydro-2',3'-dideoxy-2'-fluoro-4'-thionucleosides. AB - beta-l-2',3'-Didehydro-2',3'-dideoxy-2'-fluoro-4'-thionucleosides (beta-l-2'-F-4' S-d4Ns) have been synthesized and evaluated against HIV-1 in primary human lymphocytes. The key intermediate 8, which was prepared from 2,3-O-isopropylidene l-glyceraldehyde 1 in 13 steps, was condensed with various pyrimidine and purine bases followed by elimination and deprotection to give the target compounds, beta l-2'-F-4'-S-d4Ns (17-20 and 27-30). The antiviral activity of the newly synthesized compounds was evaluated against HIV-1 in human peripheral blood mononuclear (PBM) cells, among which the cytosine 17, 5-fluorocytosine 18, and adenine 27 derivatives showed potent anti-HIV activities (EC(50) = 0.12, 0.15, and 1.74 microM, respectively) without significant cytotoxicity up to 100 microM in human PBM, CEM, and Vero cells. The cytosine derivative 17 (beta-l-2'-F-4'-S d4C), however, showed cross-resistance to a 3TC-resistant variant (HIV-1(M184V)). Molecular modeling studies suggest that the pattern of antiviral activity, similar to that of beta-l-2'-F-d4N, stemmed from their conformational and structural similarities. The isosteric substitution of sulfur for 4'-oxygen was well tolerated in the catalytic site of HIV-1 reverse transcriptase in the wild type virus. However, the steric hindrance between the sugar moiety of the unnatural l-nucleoside and the side chains of Val184 of M184V RT in 3TC-resistant mutant HIV strains destabilizes the RT-nucleoside triphosphate complex, which causes the cross-resistance to 3TC (M184V mutant). PMID- 12540239 TI - Indanylidenes. 1. Design and synthesis of (E)-2-(4,6-difluoro-1 indanylidene)acetamide, a potent, centrally acting muscle relaxant with antiinflammatory and analgesic activity. AB - The design of rigid cyclic analogues derived from cinnamamide 1, (E)-N cyclopropyl-3-(3-fluorophenyl)prop-2-enamide, and beta-methylcinnamamide 2, (E)-N cyclopropyl-3-(3-fluorophenyl)but-2-enamide, has led to the discovery of the potent, centrally acting muscle relaxant (E)-2-(4,6-difluoro-1 indanylidene)acetamide, 17. Compound 17 also possesses potent antiinflammatory and analgesic activity. This paper describes the synthesis and the muscle relaxant, antiinflammatory, and analgesic structure-activity relationships of 17 and 67 of its analogues. Compound 17 has been taken into phase I clinical trials. PMID- 12540240 TI - Indanylidenes. 2. Design and synthesis of (E)-2-(4-chloro-6-fluoro-1 indanylidene)-N-methylacetamide, a potent antiinflammatory and analgesic agent without centrally acting muscle relaxant activity. AB - Extension of the structure-activity relationship studies that led to the discovery of the nonsedating potent muscle relaxant, antiinflammatory, and analgesic agent (E)-2-(4,6-difluoro-1-indanylidene)acetamide, 1, has given rise to (E)-2-(4-chloro-6-fluoro-1-indanylidene)-N-methylacetamide, 2. Compound 2 is a potent antiinflammatory and analgesic agent without centrally acting muscle relaxant activity. PMID- 12540241 TI - Substituted pyrrol-1-ylacetic acids that combine aldose reductase enzyme inhibitory activity and ability to prevent the nonenzymatic irreversible modification of proteins from monosaccharides. AB - Starting from the known inhibitory activity of (3-benzoylpyrrol-1-yl)acetic acid (I) and (2-benzoylpyrrol-1-yl)acetic acid (II), a series of 3-aroyl and 2,4-bis aroyl derivatives (54-75) were synthesized and tested for inhibition of aldose reductase, an enzyme involved in the appearance of diabetic complications. It was found that a number of the tested compounds exhibited considerable activity in the micromolar range. Important structural features for the potent compounds is the presence of substituents with relatively low Hammett sigma values and/or moieties which increase their overall aromatic area. The most active derivative was the [2,4-bis(4-methoxybenzoyl)pyrrol-1-yl]acetic acid (75), with potency favorably compared to known ARIs such as tolrestat, epalrestat, zopolrestat, and fidarestat. Four selected derivatives were also evaluated for their ability to interfere with the oxidative modification of serum albumin in an in vitro experimental glycation model of diabetes mellitus. All of them showed considerable activity, comparable to the known inhibitor trolox. Our results, taken together, indicate that compound 75 combines favorably two biological activities directly connected to a number of pathological conditions related to the chronic diabetes mellitus. PMID- 12540242 TI - Synthesis and biological activity of nitro heterocycles analogous to megazol, a trypanocidal lead. AB - As part of our efforts to develop new compounds aimed at the therapy of parasitic infections, we synthesized and assayed analogues of a lead compound megazol, 5-(1 methyl-5-nitro-1H-2-imidazolyl)-1,3,4-thiadiazol-2-amine, CAS no. 19622-55-0), in vitro. We first developed a new route for the synthesis of megazol. Subsequently several structural changes were introduced, including substitutions on the two rings of the basic nucleus, replacement of the thiadiazole by an oxadiazole, replacement of the nitroimidazole part by a nitrofurane or a nitrothiophene, and substitutions on the exocyclic nitrogen atom for evaluation of an improved import by the glucose or the purine transporters. Assays of the series of compounds on the protozoan parasites Trypanosoma brucei, Trypanosoma cruzi, and Leishmania donovani, as either extracellular cells or infected macrophages, indicated that megazol was more active than the derivatives. Megazol was then evaluated on primates infected with Trypanosoma brucei gambiense, including late-stage central nervous system infections in combination with suramin. Full recovery was observed in five monkeys in the study with no relapse of parasitemia within a 2 year follow-up. Because there is a lack of efficacious treatments for sleeping sickness in Africa and Chagas disease in South America, megazol is proposed as a potential alternative. The mutagenicity of this compound is at present being reevaluated, and metabolism is also under investigation prior to possible further developments. PMID- 12540243 TI - Furoyl and benzofuroyl pyrroloquinolones as potent and selective PDE5 inhibitors for treatment of erectile dysfunction. AB - Synthesis of furoyl and benzofuroyl pyrroloquinolones as potent and selective PDE5 inhibitors was reported. Their in vitro potencies in inhibiting PDE5 and selectivity in inhibiting other PDE isozymes (PDE1-4 and PDE6) were evaluated. Some of these compounds are more potent than sildenafil with better selectivity toward PDE1 and PDE6. Incorporation of solublizing groups resulted in bioavailable analogues. Selected compounds showed in vivo efficacy in anesthetized dog model for penile erection. PMID- 12540245 TI - Use of the child development review increases residents' discussion of behavioral problems. AB - OBJECTIVE: To assess whether use of a parent questionnaire, the Child Development Review (CDR), before health maintenance examinations (HMEs) increases parent identification and increases resident discussion and documentation of behavioral issues for toddler and preschool-aged children. STUDY DESIGN: Quasi-experimental design using convenience samples with comparisons before and after questionnaire implementation. SAMPLE: Children 15 to 47 months of age and their parents attending a hospital-based, primary care clinic serving a low-income population, and the residents caring for them. METHODS: Participants were enrolled into early control, concurrent control, and intervention groups. All parents completed an exit interview about 9 behavioral issues and were asked if they had discussed these issues with the doctor. Before the visit, the intervention group parents completed the CDR, which was available for the residents to review. Information from the CDR and exit interview, and resident documentation were compared for control and intervention groups. RESULTS: There were 122 intervention and 135 control group HMEs. Parents using the CDR identified more behavioral issues for their children at the exit interview (median, 2.0 vs 1.0, P =.01). The 6 issues on both the CDR and the exit interview accounted for this increase. Intervention group parents reported discussion of more behavioral issues (median, 2.0 vs 0.0, P <.001). Discussion increased for all issues. Documentation was not increased for the intervention group. CONCLUSION: Use of the CDR before HMEs increased parent identification and reported discussion of behavioral topics for toddler and preschool children. PMID- 12540246 TI - Simple feedback notes enhance specificity of feedback to learners. AB - BACKGROUND: Learners reported that physician educators give insufficient and general feedback. OBJECTIVE: This study analyzed faculty's use of 3 x 5-inch feedback notes (preprinted with "well done" or "needs improvement") for quantity and specificity of feedback. Learners evaluated these notes compared with other feedback methods. DESIGN/METHODS: Seven experienced clinician educators presented the feedback notes to learners during the 10-month trial. A carbonless duplicate was retained for qualitative analysis of content. The specificity of comments was categorized as general (no clarifiers) or detailed if it contained 1 or more clarifiers. Additionally, the learners were surveyed regarding utility of notes on the basis of characteristics of effective feedback. RESULTS: A total of 770 notes containing 1607 individual comments were presented to learners, with significantly more (P <.001) "well done" comments (69%) than "needs improvement" comments (31%). Eleven content areas emerged. The most frequently coded "well done" content areas were assessment (n = 258), treatment (n = 208), and physical examination (n = 176). The most frequently coded "needs improvement" content areas were documentation (n = 161) and expanding knowledge (n = 102). Eighty-two percent of the comments were specific, and the specificity increased significantly from 24% in the initial 4-month time frame to 46% in the last 3 months (P <.001, Pearson chi-square test). Learners agreed that feedback notes provided more constructive (94%), timely (92%), and concrete (94%) feedback when compared with other feedback methods. Learners preferred feedback given face-to face (96%) or by written notes (96%) rather than by e-mail (30%). CONCLUSIONS: Cued feedback notes provided learners with detailed feedback in a format that learners prefer. Over time, simple feedback notes lead to more specific feedback for trainees. PMID- 12540247 TI - A comparison of clinical pediatric patient encounters in university medical center and community private practice settings. AB - BACKGROUND: Growing numbers of medical students complete clerkships in community private practice (CPP) settings instead of the more traditional university-based clinics, yet few empirical studies have evaluated how setting type impacts clinical experiences, skill development, and student satisfaction. OBJECTIVE: This study compared the pediatric patient encounters seen by third-year medical students in university medical center (UMC) and CPP settings. METHODS: Third-year medical students were required to keep a log of all patients seen during their 8 week pediatric clerkship. Logbook entries were coded and then analyzed for differences in the number and distribution of the primary diagnostic categories between settings. RESULTS: CPP students reported, on the average, seeing over 3 times more patients than UMC students. The case mix distribution also differed significantly by setting. In general, CPP students reported seeing proportionately more routine illnesses, whereas UMC students reported seeing proportionately more uncommon disorders. Because CPP students saw more patients overall, they averaged more cases in almost all diagnostic categories. CONCLUSION: CPP students received more clinical patient exposure than UMC students, except for patients in a few diagnostic categories. PMID- 12540249 TI - Neurotoxic risk caused by stable and variable exposure to methylmercury from seafood. AB - OBJECTIVES: To examine whether the dose-effect relationship for developmental mercury neurotoxicity is affected by variable mercury exposure during pregnancy. METHODS: The study was based on a birth cohort of 1022 children born in the Faroe Islands between March 1986 and December 1987. Neurobehavioral performance of 917 children (90%) was assessed at age 7. Intrauterine methylmercury exposure was determined from mercury concentrations in cord blood and 2 sets of maternal hair. Complete exposure information was available for 614 children (67%). RESULTS: In children with complete exposure data, 8 of 16 neuropsychological tests showed deficits significantly associated with the cord-blood mercury concentration after confounder adjustment. Variable intrauterine exposure was suggested by disagreement between mercury concentrations in the 2 maternal hair samples. Removal of the 61 children (10%) with the greatest degree of variable exposure had a minimal effect on most exposure-effect relationships. However, the effect of the cord-blood concentration on verbal learning and memory was greater after this exclusion. CONCLUSION: The study supports previous findings from this cohort study that maternal mercury exposure during pregnancy is associated with neuropsychological deficits detectable at age 7 years and that this association is evident in women with stable exposures throughout pregnancy. Thus the association is not the result of variable exposures. PMID- 12540250 TI - Mercury exposure in an urban pediatric population. AB - OBJECTIVE: To determine the prevalence of elevated urinary mercury (Hg), as a marker of exposure, in a population of children drawn from an inner-city community with documented access to elemental mercury. METHODS: A prospective consecutive patient series was conducted from November 1998 to January 1999 at an inner-city clinic in New York. Anonymous urine specimens from subjects (aged 1-18 years) were collected in mercury-free containers, split, acidified with 1:100 hydrochloric acid, and frozen. Cold-vapor atomic absorption spectrofluorometric assays were conducted simultaneously at laboratories at the University of Washington and the New York City Department of Health. RESULTS: We enrolled 100 children (mean age 9.4 years; 62% male; 55% Hispanic; and 43% African American). Assay results from both laboratories were strongly correlated (r = 0.8, P <.0001). Mean urinary Hg was 1.08 +/- 1.82 microg/L. The 95th percentile for urinary Hg was 2.8 microg/L (range 0.2 to 11.7 microg/L). Five subjects had Hg levels above 5 microg/L. CONCLUSION: We found that 5% of subjects had unsuspected elevated urinary Hg levels. This finding, in a group of inner-city minority children, strongly supports the need for further investigation of the sources of mercury exposure in this population. PMID- 12540251 TI - Prevention of lead toxicity in US children. AB - During the past 2 decades, the proportion of US children who have blood lead concentrations of 10 microg/dL or higher declined by over 80% after the elimination of leaded gasoline and lead solder from canned foods, and a ban on leaded paint used in housing and other consumer products. Fatalities and symptomatic lead poisoning are now rare. Residential lead hazards, which are exceedingly difficult to control, are currently the major source of lead intake for children. Undue lead exposure has retreated into 2 major risk groups; impoverished children who live in older, poorly maintained rental housing and more affluent children who live in older housing undergoing renovation. Despite the dramatic decline in children's blood lead levels, lead toxicity remains epidemic among impoverished children who live in older rental housing, especially those who live in the northeastern and midwestern regions of the United States. There are increasing data linking lead exposure with other systemic effects including delinquency, dental caries, and learning problems. Moreover, there is evidence indicating that there is no discernible threshold for lead-associated cognitive deficits. Thus, it is increasingly important to shift our efforts toward the primary prevention of childhood lead exposure from residential hazards. This article reviews the epidemiology and control of childhood lead exposure, focusing especially on steps necessary to shift toward primary prevention. PMID- 12540252 TI - Severe lead poisoning in pregnancy. AB - BACKGROUND: Lead freely crosses the placenta. Consequently, gestational lead poisoning is not only harmful to the woman but also to the developing fetus, invariably producing congenital lead poisoning. The scope and consequences of severe lead poisoning in pregnancy (blood lead level > or =45 microg/dL) have not been well characterized. METHODS: We reviewed our experience in the management of women with severe gestational lead poisoning. Additionally, we reviewed the literature on this disorder in an effort to identify patterns in etiology and outcome. RESULTS: Over a 3-year period treatment was provided to 7 severely lead poisoned women. A 25-year review of the medical literature identified an additional 8 cases. Among these 15 women, 70% were Hispanic, all of whom developed lead poisoning from the ingestion of soil, clay, or pottery ("tierra"). Other sources of lead poisoning were paint chip ingestion (n = 2), household renovation, and use of a complementary-alternative medication (bone meal). Lead poisoning was discovered in the third trimester in 12 (86%) subjects after the women presented with subtle but characteristic findings of severe lead poisoning, including malaise, anemia, or basophilic stippling on blood smear; one woman was identified when she presented after a generalized seizure, having a blood lead level of 104 microg/dL. Five women received chelation therapy during pregnancy with CaNa(2) EDTA, dimercaprol, or succimer. At delivery mean maternal blood lead level was 55 microg/dL, whereas mean neonatal lead level was 74 microg/dL (P =.009). Thirteen neonates underwent chelation, all within the first 28 days of life. No infant in the current series had an identifiable birth defect. CONCLUSIONS: On the basis of this experience we conclude that severe lead poisoning in pregnant women has the following characteristics: 1) it most often occurs because of intentional pica, 2) its presenting features are subtle, often consisting only of malaise and anemia, and 3) blood lead levels in the neonate are higher than simultaneous maternal lead levels. PMID- 12540253 TI - Meconium as a biological marker of prenatal exposure. PMID- 12540254 TI - The impact of climate change on child health. AB - Human activity has contributed to climate change. The relationship between climate and child health has not been well investigated. This review discusses the role of climate change on child health and suggests 3 ways in which this relationship may manifest. First, environmental changes associated with anthropogenic greenhouse gases can lead to respiratory diseases, sunburn, melanoma, and immunosuppression. Second, climate change may directly cause heat stroke, drowning, gastrointestinal diseases, and psychosocial maldevelopment. Third, ecologic alterations triggered by climate change can increase rates of malnutrition, allergies and exposure to mycotoxins, vector-borne diseases (malaria, dengue, encephalitides, Lyme disease), and emerging infectious diseases. Further climate change is likely, given global industrial and political realities. Proactive and preventive physician action, research focused on the differential effects of climate change on subpopulations including children, and policy advocacy on the individual and federal levels could contain climate change and inform appropriate prevention and response. PMID- 12540255 TI - Children's environmental health: one year in a pediatric environmental health specialty unit. AB - BACKGROUND/OBJECTIVE: As a result of an increasing desire among physicians and parents for clinical centers that can evaluate children with known or suspected exposures to environmental toxicants, a network of federally funded "pediatric environmental health specialty units" has recently been created. This descriptive study profiles the children seen in one unit of this program. SETTING: A New England, university-affiliated Pediatric Environmental Health Center (PEHC). METHODS: Review and analysis of all children seen in the PEHC in calendar year 1999. RESULTS: Over the course of the year, 281 children made 863 visits to the PEHC. Presenting complaints fell into 4 major categories: new visit for management of lead intoxication (n = 248), return visit for management of lead intoxication (n = 569), new visit for evaluation of exposure to an environmental toxicant other than lead (n = 33), and return visit for the management of exposure to a non-lead toxicant (n = 13). Among those children with new visits for a non-lead toxicant, the most common chief complaints were exposure to solvent-contaminated water (n = 7), pesticide exposure (n = 6), illness associated with proximity to a hazardous waste site (n = 6), autism from suspected mercury intoxication (n = 4), and evaluation of school-induced, building-related illness ("sick school syndrome")(n = 4). Eleven children had autism or pervasive developmental delay. Families traveled distances as great as 450 kilometers for evaluation by a pediatric environmental health clinical specialist. Every child was evaluated by a pediatrician with subspecialty training in medical toxicology. Environmental investigation of air, water, paint, dust, or land was conducted for all except 4 children (all foreign-born adoptees). Therapeutic interventions included chelation therapy, relocation to a safe environment, removal from school, and termination of chelation therapy that had been initiated by another practitioner. Third-party payors provided full reimbursement for all visits. CONCLUSIONS: The chief complaints of the children brought to pediatric environmental health specialty units are diverse, involving exposures to a wide range of toxicants from all environmental media (air, water, soil, and food). Parents desiring such an evaluation must often travel extensive distances, suggesting the need for a broader network of such centers. Third-party payors and health maintenance organizations are willing to provide full reimbursement for these services. PMID- 12540256 TI - Pediatric environmental health education: a survey of US pediatric residency programs. PMID- 12540257 TI - Pediatric environmental health competencies for specialists. AB - BACKGROUND: Because environmental health problems are complex and require specialty training, the Ambulatory Pediatric Association initiated a 3-year postgraduate fellowship in Pediatric Environmental Health. OBJECTIVE: To develop competencies for the specialty of Pediatric Environmental Health and appropriate measures (performance indicators) for the achievement of these competencies. METHODS: The President of the Ambulatory Pediatric Association appointed a 6 member Fellowship Oversight Committee to guide the development of the Fellowship Program and to draft competencies for fellows in Pediatric Environmental Health. The Committee developed a list of proposed competencies for graduates of Pediatric Environmental Health fellowships. These were skills identified as very important for a specialist to have for minimal competency in the practice of pediatric environmental health. RESULTS: Twenty-seven Pediatric Environmental Health competencies are proposed. The competencies are presented from 3 separate perspectives: academic, individual patient care, and community advocacy. Each competency has a list of suggested performance indicators. CONCLUSION: These competencies are intended to assist in structuring the training experience, achieving consensus with respect to expectations of fellows and faculty, providing opportunities for fellows to assess their own needs or gaps in training, and identifying the expertise of fellowship graduates to potential employers. PMID- 12540258 TI - Emerging targets for antimalarial drugs. AB - The absence of an effective vaccine against malaria and the ability of the parasite to develop resistance to known antimalarial drugs makes it mandatory to unravel newer drug targets with a view to developing newer pharmacophores. While conventional targets such as the purine, pyrimidine and folate pathways are still being investigated in the light of newer knowledge, a new opportunity has emerged from an understanding of certain unique features of the parasite biology. These include the food vacuole, haemoglobin catabolism, haeme biosynthesis, apicoplasts and their metabolism as well as macromolecular transactions, import of host proteins, parasite induced alterations in the red cell surface and transport phenomena. This review seeks to emphasise the new and emerging targets, while giving a brief account of the targets that have already been exploited. PMID- 12540259 TI - Targeting HIV-1 integrase. AB - Human immunodeficiency virus Type 1 (HIV-1) integrase is an essential enzyme for the obligatory integration of the viral DNA into the infected cell chromosome. As no cellular homologue of HIV integrase has been identified, this unique HIV-1 enzyme is an attractive target for the development of new therapeutics. Treatment of HIV-1 infection and AIDS currently consists of the use of combinations of HIV 1 inhibitors directed against reverse transcriptase (RT) and protease. However, their numerous side effects and the rapid emergence of drug-resistant variants limit greatly their use in many AIDS patients. In principle, inhibitors of the HIV-1 integrase should be relatively non-toxic and provide additional benefits for AIDS chemotherapy. There have been many major advances in our understanding of the molecular mechanism of the integration reaction, although some critical aspects remain obscure. Several classes of compounds have been screened and further scrutinised for their inhibitory properties against the HIV integrase; however, there are currently no useful inhibitors available clinically for the treatment of AIDS patients. This review describes the current knowledge of the biological functions of the HIV-1 integrase and reports the major classes of integrase inhibitors identified to date. PMID- 12540260 TI - Microbial genomics - new targets, new drugs. AB - Genomics has changed our view of the biological world in the past decade, providing both new information and new tools to characterise biological systems. Over 100 microbial genomes - including many of substantial clinical importance - have been fully or partially sequenced, pushing the search for novel antimicrobial compounds into the post-genomic era. Genomic information and associated new technologies have the potential to revolutionise the drug discovery process. Genomic methods have created a wealth of potential new antimicrobial targets; strategies are evolving to provide validation for these targets before chemical inhibitors are identified. The ability to obtain large amounts of purified target proteins and advances in X-ray crystallography have caused significant increases in available protein structures, which may foreshadow an increased effort in structure-based drug design. The post-genomics strategies used in antimicrobial drug discovery may have application for small molecule drug discovery in numerous therapeutic areas. PMID- 12540261 TI - Glutathione S-transferases as emerging therapeutic targets. AB - Glutathione S-transferases (GST) represent a large family of Phase II detoxification enzymes widely expressed in animals and plants. These enzymes catalyse the conjugation of glutathione with some endogenous molecules and a broad range of exogenous substrates including various anticancer drugs. Due to high expression of GSTs in tumours when compared to normal tissues and their high level in plasma from cancer patients, these enzymes are considered to be cancer markers. Their involvement in resistance to anticancer drugs and an inverse correlation between expression and prognosis in many tumours provided a rationale for the design of inhibitors and prodrugs to enhance therapeutic index. The first generation of GST inhibitors included ethacrynic acid and showed promising potentiating activity in vitro but lack of isoenzyme specificity and diuretic side effects restricted clinical use. Novel GST inhibitors include glutathione analogues and demonstrate better specificities with fewer limiting toxicities. One lead compound is a potent inhibitor of the GSTP1-1 isoform in both cell lines and animal models. A GSTP1-1 activated prodrug has also been developed. Testing of the preclinical and clinical efficacy of these agents is presently in progress. Their rational design provides a promising new approach to targeting tumour-specific characteristics in a manner consistent with improving therapeutic index. PMID- 12540262 TI - Dendritic cells: immunological features and utilisation for tumour immunotherapy. AB - The prospect of developing 'magic bullets' to attack tumour cells has been a goal of biologists for decades. Abundant experimental and clinical observations demonstrating that an effective specific immune response may engender tumour regression has prompted efforts to find an immunotherapeutic approach to this problem. The most important arm of cellular immunity for such responses appears to be cytotoxic T-lymphocytes (CTL) which can recognise antigen on virtually all cell types and which are key to the elimination of virally-infected cells. The specific activation and maintenance of activity of these cells is therefore the major goal of designing a therapeutic cancer vaccine. Advances in our understanding of the role of dendritic cells (DC) in priming and modifying immune responses suggest that they should be potent adjuvants for vaccination. The use of antigens targeted to the major histocompatibility complex (MHC) molecules expressed on these cells as an approach to tumour immunotherapy has already been tested in the treatment of many malignancies, and recent findings shed light on additional directions through which their efficacy may be improved. PMID- 12540263 TI - Glycine transport inhibitors as potential antipsychotic drugs. AB - Current antipsychotic drugs are only partially effective in treating schizophrenia and there is a clear need to develop better therapies. An alternative approach to develop new antipsychotics has come from the NMDA receptor hypofunction model for schizophrenia. It has been hypothesised that stimulation of NMDA receptors with glycine site agonists may be therapeutic, and a number of clinical trials of glycine together with standard antipsychotic drugs have been recently been conducted. Modest improvements in negative symptoms have been reported in some studies but a potentially more effective treatment is to use inhibitors of the GLYT1 subtype of glycine transporters. Expression of GLYT1 within the brain correlates with NMDA receptor expression patterns and it has been suggested that GLYT1 may regulate synaptic glycine concentrations. With the development of selective and potent non-transported inhibitors of GLYT1 it should be possible to elevate synaptic glycine concentrations more effectively and thereby to increase NMDA receptor activity. Recent in vitro studies demonstrate that the glycine transport inhibitor, N[3-(4-fluorophenyl)-3-(4'-phenylphenoxy)] propylsarcosine, enhances NMDA receptor activity and the use of this class of compounds in clinical studies is eagerly awaited. PMID- 12540264 TI - Connective tissue growth factor: an attractive therapeutic target in fibrotic renal disease. AB - Despite diverse initiating insults, glomerulosclerosis and tubulointerstitial fibrosis are pathological features common to most forms of progressive renal disease. Control of systemic hypertension and blockade of the renin-angiotensin system ameliorate the rate of progression of chronic renal disease; however they generally fail to completely arrest the scarring process. While the chain of events leading to scarring are still being defined, TGF-beta is a cytokine that plays a pivotal role in the pathogenesis of glomerulosclerosis and tubulointerstitial fibrosis [1]. Given the pleiotropic effects of TGF-beta, significant attention has focused on the potential of its downstream mediators as therapeutic targets. Connective tissue growth factor (CTGF) is a member of the CCN gene family, which includes CyR61 (cysteine rich 61), Nov (Nephroblastoma overexpressed) and the WISP family (for review see [2,3,4]). These immediate early genes coordinate complex biologic processes during differentiation and tissue repair [5]. Increased expression of CTGF has been detected in experimental and human renal fibrosis where it correlates with glomerulosclerosis and the degree of tubulointerstitial fibrosis [6]. In these settings CTGF expression is regulated at least in part by TGF-beta. This review details the biology of CTGF with specific reference to its potential as a therapeutic target in renal fibrosis. PMID- 12540265 TI - Targeting DNA gyrase. AB - The discovery of the peptide DNA gyrase inhibitor microcin B17 (MccB17) in the early 1990s provided a new tool and hope for a novel peptide-based chemical starting point for a new generation of DNA gyrase inhibitors but the definitive mechanism-of-action of MccB17 has remained unknown. This research report [1], by one of the foremost laboratories in this discipline in the world, provides definitive data on the mode of inhibition of MccB17 and possibly opens the door for additional semisynthetic analogue synthesis based on the MccB17 chemotype. In addition, this unique peptide DNA gyrase inhibitor provides a contrast in activity versus quinolones, Ca(2+)-mediated inhibition/cleavage and the bacterial toxin/peptide CcdB. PMID- 12540266 TI - Target validation and functional analyses using antisense oligonucleotides. AB - The human genome project (HGP) has been described as the single most important project in biology and the biomedical sciences to date. In February 2001, the efforts of the HGP resulted in the publication of a 'working draft' of the entire human genome and it is expected that final sequencing and annotation of the genome will be completed by 2003. Researchers are now focusing efforts on the identification of the function of the reported 30,000 human genes. During the past few years, antisense oligomers have been widely used as potent tools for functional genomics and drug target validation. This article describes the emerging and established antisense technologies that will be used to continue the efforts to unlock the function of the human genome and to discover novel drug targets for the treatment of human diseases. PMID- 12540267 TI - Chemokine receptors: emerging opportunities for new anti-HIV therapies. AB - The chemokine receptors CCR5 and CXCR4 are G-protein coupled receptors (GPCRs) of the immune system and the major co-receptors required for entry of HIV into CD4(+) target cells. CCR5 is critical for both human immunodeficiency virus (HIV) disease transmission and progression, whereas CXCR4 may be very important in late stages of disease. Additional co-receptors have been shown to function under certain conditions in vitro but evidence of supporting roles in HIV disease is currently lacking. The sheer number of co-receptors potentially used by HIV and the complexity of co-receptors usage are major challenges confronting usage of these molecules as drug development targets. Balanced against this, is a long history of success by the pharmaceutical industry in developing small molecule antagonists for many other classes of GPCRs. In this review, we discuss the current state of understanding of the co-receptor-based antiviral agents designed to block viral entry. The therapeutic potential of this field will be judged from future studies on the efficacy of these novel inhibitors in clinical trials. The data so far obtained from a number of studies point to the potential clinical use of this emerging class of therapeutic agents. Here we review current progress in co-receptor-based antiretroviral drug development and discuss the potential advantages and disadvantages of this approach. PMID- 12540268 TI - The Type III secretion system of Gram-negative bacteria: a potential therapeutic target? AB - Several pathogenic Gram-negative bacteria, including Salmonella, Shigella, Yersinia, Pseudomonas aeruginosa and enteropathogenic Escherichia coli harbour a complex attack system called 'Type III secretion' which is, in every case, an essential virulence determinant. This system, activated by contact with an eukaryotic cell membrane, allows bacteria to inject bacterial proteins across the two bacterial membranes and the eukaryotic cell membrane, to reach the cell's cytosol and destroy or subvert the host cell. The Type III virulence mechanism consists of a secretion apparatus, made up of about 25 proteins, and a set of effector proteins released by this apparatus. The mechanism of protein secretion is highly conserved among the different bacteria, although they cause a variety of diseases with different symptoms and severities, from fatal septicaemia to mild diarrhoea or from fulgurant diarrhoea to chronic infection of the lung. This review focuses on the proteins that make up the secretion machinery and examine if it could be a potential target for novel antimicrobials. PMID- 12540269 TI - Human beta-secretase and Alzheimer's disease. AB - Alzheimer's disease (AD) is a huge unmet medical need. Studies of the brain pathology and genetics of familial forms of AD have led to the amyloid cascade hypothesis, stating that Abeta42, a proteolytic breakdown product of the large amyloid precursor protein, plays an early and critical role in AD pathogenesis. Abeta42 generation requires two proteases, beta- and gamma-secretase, and inhibition of these enzymes is a key focus of AD drug development. Progress in this area has been slow because these enzymes were not identified. Using an expression cloning strategy we have identified a novel membrane bound aspartic protease, BACE1 and demonstrated that it exhibits all known properties of beta secretase. The enzyme has been characterised in detail. The crystal structure, which is critical for rational inhibitor design, has been solved and shown to be very similar to that of other pepsin family members. Our most recent BACE1 knockout studies show that BACE1 is critical for Abeta generation; however the knockout mice show an otherwise normal phenotype, raising the possibility that therapeutic BACE1 inhibition could be accomplished without major mechanism based toxicity. PMID- 12540270 TI - Endocannabinoids Part II: pathological CNS conditions involving the endocannabinoid system and their possible treatment with endocannabinoid-based drugs. AB - Changes in the levels of either the cannabinoid CB(1) receptors or of their endogenous ligands, anandamide and 2-arachidonoylglycerol, appear to be casual or consequential in many neurological disorders. Several examples of how such diseases may be treated by substances capable of selectively manipulating endocannabinoid levels and action are presented, using animal models of neuropathological conditions, such as motor disorders, multiple sclerosis, neuronal damage, chronic and inflammatory pain, anorexia, cachexia and motivational disturbances. These examples indicate that new therapeutic agents, lacking the undesirable psychotropic side effects of Cannabis, may be developed from current studies on the endocannabinoid system. PMID- 12540271 TI - Telomerase - strategies to exploit an important chemotherapeutic target. AB - Telomeres, unique protein-DNA complexes located at the chromosome ends, have important functions involving both DNA protection and cellular signalling. Telomere structure is very dynamic yet tightly controlled. One important factor is the presence of telomerase, a telomere-specific DNA polymerase activated in a majority of cancer cells. Cancer and normal cell telomeres may have dissimilar structures due to variances in telomere length, telomerase activity and levels of telomere binding proteins. In designing compounds to strictly target cancer cells, these distinctions should be investigated. Much of the recent focus has been on the development of highly effective telomerase inhibitors. Another novel group of small molecules target telomere DNA, thereby disrupting both telomerase activity and telomere structure. This class of compounds should have an immediate impact on cell growth and viability. Since many molecular characteristics of telomeres are unknown, small molecules should also be useful in probing differences in telomere dynamics unique to cancer cells. PMID- 12540272 TI - Mechanisms of MHC class I-restricted antigen presentation. AB - The vertebrate immune system monitors whether an organism is invaded by pathogens. Therefore, each cell has to prove itself as healthy. This is achieved by presenting fragments of intracellular protein degradation products on the surface, i.e., each cell displays peptides on specialised proteins known as major histocompatibility complex (MHC) class I proteins. A displayed peptide has to pass certain constraints before its presentation: It has to be excised out of a protein, translocated into the endoplasmic reticulum (ER) and fit into the binding groove of a MHC molecule. In theory, alteration of the cellular protein profile by mutation or infection should force pathogen-specific T-cells to take action via recognition of foreign peptide bound to MHC class I molecules on the cell surface. Unfortunately, pathogens and tumours have evolved many ways to affect antigen presentation and to escape from immune response. Understanding the exact mechanisms of antigen presentation, i.e., protein cleavage and peptide binding by MHC molecules, would allow their manipulation by drugs and lead to the re-establishment of the correct antigen presentation pathway. This review will summarise current knowledge of the mechanisms of antigen presentation and discuss putative targets for therapeutic treatment as well as for vaccination strategies. PMID- 12540273 TI - Emerging molecular targets for the treatment of pre-eclampsia. AB - Pre-eclampsia (PE) is a pregnancy-specific syndrome that is a principal cause of maternal morbidity and mortality, accounting for almost 15% of pregnancy associated deaths. It is also one of the major causes of iatrogenic prematurity among new born babies, placing a heavy burden on both prospective parents and on the health service. The mild form of PE most commonly presents with the features of maternal hypertension and proteinuria but can swiftly and unpredictably become severe with many extensive complications, which can involve the maternal liver, kidneys, lungs, blood and platelet coagulation and nervous systems. These clinical problems normally only become apparent in the second half of pregnancy but are believed to start during the first trimester. The diverse symptoms of PE have made it a difficult disease not only to define but also to identify a causative agent for the symptoms. It has therefore proved difficult to develop specific drugs that can be used to manage the condition in the clinic. Therapeutic intervention so far has been primarily aimed at combating the two main complications of PE - the hypertension and seizures. Current therapies are widely recognised as inadequate. This review examines the complex pathological mechanisms believed to be responsible for the multi-system complications of PE. It highlights current findings that exhibit the potential to target these effects with the aim of either preventing or altering the course of this life-threatening disease of pregnancy. PMID- 12540274 TI - Resistance to anti-peptide deformylase drugs. AB - Recent work has assessed the potential of peptide deformylase (PDF) as a target for broad spectrum antibacterial agents. By using a number of approaches, including proteomics, researchers at Roche have shown that the molecules they had selected in vitro were able to target PDF in vivo. However, the authors, having observed resistance occurring at a rather high frequency and on the basis of the recent discovery of a deformylase homologue in humans, suggest that PDF 'may not be an optimal target for broad spectrum antibacterial agents'. We link these data to results published by other laboratories and conclude that PDF deserves to still be considered a valuable target for new antibiotics. PMID- 12540275 TI - Cytokine signalling and disease. AB - Cytokines mediate their response via cell surface receptors that in turn activate intracellular signalling pathways and lead to gene activation, cell proliferation and differentiation. Many recent studies have shown that cytokine and cytokine receptor pathways are frequently mutated in disease, thus shedding light on the generation of the inflammatory response, specific immunity and mechanisms of haematopoiesis. Many approaches are being used to translate this basic research into successful therapies and although host immune responses involve many different cells and crucial pathways, modulation of therapeutic responses can be induced or inhibited by, targeting a single cytokine. This review summarises current knowledge of cytokine pathways in disease and the use of cytokine- or receptor-directed therapy to exploit the immune response to disease. PMID- 12540276 TI - Regulation of microglia - potential new drug targets in the CNS. AB - Microglia respond to any disturbance in the CNS which poses a threat to physiological homeostasis. Although these responses are secondary, mainly to neuronal alterations, the way the microglial response evolves in many situations promotes further damage to the CNS. The list of clinical conditions in which this situation is a major problem is continuously growing and includes neurodegenerative diseases, stroke, trauma, demyelinating disorders and neuropathic pain. The significance of microglia for the pathogenesis of neurological and neuropsychiatric conditions has led to a rapidly expanding search for therapeutic possibilities to regulate microglial activity. As will be clear from this review, treatments which are currently available appear to offer some positive effects but are still far from satisfactory. A major challenge is to understand the mechanisms that determine whether activated microglia will develop into a cytotoxic or a cytoprotective component. PMID- 12540277 TI - Metabotropic glutamate receptors: targets for therapy of cerebral ischaemia. AB - Several pathophysiological processes are involved in the progression of the cerebral ischaemic injury, among which are inflammation, peri-infarct depolarisation, apoptosis and excitotoxicity. Overactivation of glutamate receptors, both ionotropic and metabotropic, constitutes the central step in the excitotoxic cascade of events leading to neuronal cell death following acute brain ischaemia. Owing to their peculiar characteristics of modulatory receptors and due to their wide molecular and biological diversity, metabotropic glutamate receptors constitute an attractive target for the development of potential neuroprotective agents. Recent achievements in developing novel chemical entities and in the characterisation of the physiological and pathological role of individual metabotropic glutamate receptors in postischaemic degeneration will be reviewed. PMID- 12540278 TI - Insights into the structure and function of 5-HT(2) family serotonin receptors reveal novel strategies for therapeutic target development. AB - 5-HT(2) family serotonin receptors, principal sites of action of serotonin in the brain, represent major molecular targets for drugs used in treating a variety of diseases including schizophrenia, depression, anxiety, eating disorders, obsessive-compulsive disorder, chronic pain conditions and obesity. The 5-HT(2) family of receptors has three members: 5-HT(2A), 5-HT(2B) and 5-HT(2C). Therefore, it is likely that subtype-selective compounds will be needed to avoid serious side effects and to enhance therapeutic indices. Unfortunately, recent insights into the structure and function of 5-HT(2A) receptors have revealed that structurally-diverse agonists and antagonists have distinct modes of interacting with 5-HT(2A) receptors, complicating efforts at structure-based drug-design. These distinct binding modes would not have been predicted based on conventional structure-activity relationships or static docking models. Fortunately, these complicated binding modes can be predicted and simulated using molecular dynamics, allowing for the possibility of structure-based drug design. Thus, provided appropriately sophisticated drug design strategies are employed, it is likely that uniquely valuable medications will result which could have great potential for treating a variety of mental and physical illnesses. PMID- 12540279 TI - Superoxide dismutase: an emerging target for cancer therapeutics. AB - Superoxide dismutase (SOD) is a critical enzyme responsible for the elimination of superoxide radicals and is considered to be a key anti-oxidant in aerobic cells. Cellular consumption of oxygen is essential for oxidative phosphorylation during ATP generation in the mitochondria, yet this cellular metabolism also leads to the production of reactive oxygen species (ROS), including the superoxide radical (O(2)(*)(-)) and hydrogen peroxide (H(2)O(2)). Accumulation of ROS results in cellular oxidative stress and, if not corrected, can lead to the damage of important biomolecules such as membrane lipids, proteins and DNA. Prolonged accumulation of high levels of free radicals in cells may cause irreversible cellular injury and ultimately result in cell death. Since SOD is the key enzyme in the first metabolic step of superoxide elimination, deficiency in SOD or inhibition of the enzyme activity may cause severe accumulation of O(2)(*)(-) in cells and lead to cell death. Thus, inhibition of SOD may provide a novel way to kill cancer cells. Due to dysfunction in the regulation of cell growth, cancer cells are active in energy metabolism, and thus produce high levels of O(2)(*)(-) and other ROS and are under constant oxidative stress. This may render the malignant cells more dependent on SOD to eliminate the toxic superoxide radicals and thus potentially more sensitive to SOD inhibitors. It is a plausible hypothesis that inhibition of SOD may preferentially kill malignant cells through a free radical-mediated mechanism. This article will review evidence that suggests SOD as an emerging therapeutic target for cancer treatment. The relevant clinical implications and potential risk will also be discussed. PMID- 12540280 TI - New developments in the urokinase-type plasminogen activator system. AB - The urokinase-type plasminogen activator (uPA) system plays a central role in control of cell surface proteolysis and extracellular matrix degradation. Components of this system are upregulated in a wide variety of human cancers and high levels of these proteins predict more rapid relapse and shorter survival. Recently, additional complexities in this system have been recognised, especially with regard to the roles of plasminogen activator inhibitor-1 (PAI-1), the urokinase receptor (uPAR) and urokinase:uPAR complexes. PAI-1 has been shown to play a major role in the process of pathological angiogenesis. The uPAR is involved as a key player both in proteolysis and cellular adhesion, where it is both an adhesion receptor itself for vitronectin and interacts with and modifies signalling from integrins. In addition, binding of uPA to the receptor can induce intracellular signalling via a number of different pathways, including integrins and G proteins. These new developments lead to a number of novel targets for drug discovery beyond better established enzyme inhibitors and receptor antagonists. PMID- 12540281 TI - New approaches for treatment of diabetic nephropathy: the endothelium as a target for drug discovery. AB - Diabetic nephropathy is the leading cause of end-stage renal disease (ESRD). There are currently no therapeutic Hyperlipidaemiants which directly intervene in the pathogenesis of diabetic nephropathy. The mechanisms behind development of diabetic nephropathy are complex and not completely understood. The hyperglycaemia, hyperlipidaemia and hyperinsulinaemia found in diabetics might all act on the kidney endothelium to induce expression of genes important in kidney dysfunction. We propose that targeting kidney endothelial gene expression may provide a new approach for control of kidney dysfunction. PMID- 12540282 TI - Emerging therapeutic targets in nitric oxide-dependent cardiac disease. AB - Nitric oxide (NO) is a free radical gas that plays paracrine/autocrine and intracrine roles in maintaining physiological cardiovascular performance. In the coronary circulation, NO mediates endothelium-dependent vasodilator responses to shear stress and agonist-induced responses to neurohumoral stimulation. In the heart, NO modulates myocardial relaxation, beta-adrenergic responses, mitochondrial respiration and substrate metabolism and excitation-contraction coupling. Endothelial dysfunction and the resulting decrease in the production, bioavailability and/or second messenger response-coupling has been implicated in coronary artery disease and complications associated with restenosis following coronary angioplasty, stent placement and coronary artery bypass grafting (CABG). However, there are a number of pathophysiological conditions (ischaemia reperfusion, cardiac transplant rejection, myocarditis, sepsis) in which unregulated overproduction of NO and other reactive oxygen species (ROS) results in deleterious effects on cardiac function. Given the importance of NO in cardiac physiology/pathophysiology it may serve as a potential target for interventions aimed at deterring therapeutic failures of percutaneous or surgical treatments of cardiac disease as well as serving as a primary medical intervention. This review will examine the function of NO in mediating/modulating cardiac function, stressing the concept that, depending on the milieu, NO has the potential to exert either beneficial or deleterious effects on cardiac function. Moreover, this review will summarise studies in laboratory models and human studies in which NO activity, production, availability, or second messenger activation has been enhanced or inhibited in order to provide new insight for future targeting of this system for drug development. PMID- 12540283 TI - Cyclin-dependent kinases and stroke. AB - Cyclin-dependent kinases (CDKs) are a group of enzymes predominately known for their role in cell cycle regulation in proliferating cell types. Increasing evidence, however, suggests that CDKs also promote death in neurones. These observations have lead to the notion that CDKs may serve as a therapeutic target for neuropathological conditions such as stroke. Accordingly, in this review, we will examine the evidence which indicates a role for CDKs in neuronal death and evaluate the potential of CDK inhibitors as a therapeutic target for stroke. PMID- 12540284 TI - The transmissible spongiform encephalopathies: pathogenic mechanisms and strategies for therapeutic intervention. AB - Primary neurodegenerative diseases tend to be intractable and largely affect the elderly. There is rarely the opportunity to identify individuals at risk and the appearance of clinical symptoms usually signifies the occurrence of irreversible neurological damage. This situation describes sporadic Creutzfeldt-Jakob disease which occurs world-wide, affecting one person per million per annum. The epidemic of bovine spongiform encephalopathy in the UK in the 1980s and the subsequent causal appearance of variant Creutzfeldt-Jakob disease in young UK residents in the 1990s has refocused attention on this whole group of diseases, known as the transmissible spongiform encephalopathies or prion diseases. The potentially lengthy incubation period of variant Creutzfeldt-Jakob disease, including perhaps an obligate peripheral phase, prior to neuroinvasion, marks variant Creutzfeldt Jakob disease out as different from sporadic Creutzfeldt-Jakob disease. The formal possibility of detecting individuals infected with the bovine spongiform encephalopathy agent during this asymptomatic peripheral phase provides a strong incentive for the development of therapies for transmissible spongiform encephalopathies. This review focuses on recent advances in the understanding of the pathogenesis of these diseases, with particular reference to in vitro and animal model systems. Such systems have proved invaluable in the identification of potential therapeutic strategies that either specifically target the prion protein or more generally target peripheral pathogenesis. Furthermore, recent experiments in animal models suggest that even after neuroinvasion there may be pharmacological avenues to explore that might retard or even halt the degenerative process. PMID- 12540285 TI - Emerging therapeutic targets in bipolar mood disorder. AB - Bipolar mood disorder is a chronic, severe and life-threatening psychiatric illness, whose underlying pathophysiology is still obscure. Lithium is the mainstay of treatment for this illness, with robust acute antimanic and long-term prophylactic effects. Over the past decade, valproate has been another medication shown to have possibly similar mood-stabilising properties to lithium, in double blind controlled trials. Nonetheless, among patients suffering from bipolar disorder, a substantial percentage appears to respond poorly to currently available pharmacological therapies, including lithium, valproate, carbamazepine and other newer compounds, clearly demonstrating that there is a substantial need for improved therapeutic agents. Very significant effort has been made in the past several years to elucidate the cellular mechanisms by which lithium and valproate produce their therapeutic effects. The available evidence points to a modulatory action of these compounds over multiple neural biochemical pathways and most investigations have found relevant actions of mood stabilisers on intracellular signal transduction mechanisms. Moreover, it has been shown in recent years that lithium and valproate lead to long-term changes in neural plasticity, with eventual neurotrophic and neuroprotective effects. Although these actions are not fully understood, stimulation of transcription factors and effects on gene expression are potentially involved. The search for the mechanisms of action of well-established mood-stabilisers has helped to reveal promising molecular targets to test novel therapeutic approaches. This review will examine the current investigations on the diverse biochemical and molecular pathways regulated by either lithium or valproate and highlight the potential cellular targets for the development of novel mood stabilisers. PMID- 12540286 TI - Emerging targets for the treatment of depressive disorder. AB - New agents offering novel mechanisms of action are required in the treatment of depressive disorder. Established agents targeting monoamine systems are unsatisfactory because of full and partial treatment resistance, delay in the onset of their effect and the occurrence of side effects. The monoamine hypothesis of depression is now recognised to provide an incomplete explanation of the pathophysiology of depression. New theories have recently developed and new targets for treatment have emerged. We briefly review some important candidate systems and therapeutic targets in depression: the hypothalamic pituitary-adrenal axis (HPA) and the glucocorticoid and corticotrophin-releasing factor receptors, synaptic plasticity and neurotrophins and the N-methyl-D aspartate (NMDA) receptor. The putative role of the neuropeptides substance P and neuropeptide Y, the nicotinic system and the potential therapeutic benefits of cannabinoids are also reviewed. Vagal nerve stimulation (VNS) and transcranial magnetic stimulation, serendipitous advances in treatment, are discussed briefly. PMID- 12540287 TI - The biology and physiology of somatostatin receptors. AB - The biology and therapeutic application of somatostatin and its receptors are reviewed. The focus is on recent literature and patents, especially with regard to the specific function of each somatostatin receptor subtype. Detailed mechanisms mediating the effects of somatostatin and its analogues remain to be elucidated. Nevertheless, progress is being made towards a clear picture of the cellular signalling and physiological changes regulated by somatostatin and its receptors. PMID- 12540288 TI - Drug targets in Menkes disease - prospective developments. AB - Menkes disease (MNK) is an X-linked recessive disorder characterised by a copper transporting ATPase defect. In the affected cells, copper transport from the cytosol to the Golgi apparatus is disturbed, resulting in a reduction of copper efflux. Orally-administered copper, which accumulates in the intestine, cannot be absorbed and thus a copper deficiency arises. The characteristic features of MNK are progressive neurological degeneration, connective tissue disorders and hair abnormalities, which are caused by a reduction in the activity of several copper dependent enzymes, due to concomitant copper deficiency. Subcutaneous injections of copper-histidine complex, which currently forms the accepted mode of treatment, prevent the neurological degeneration in some patients when the treatment is initiated soon after birth. However, when the treatment is started later, the neurological degenerative processes are not prevented. Moreover, the treatment does not improve the connective tissue disorders that are caused by the low activity of lysyl oxidase. In order to solve these problems, a form of the treatment aimed at delivering copper into the Golgi apparatus should be studied. An attempt is made in this review to present what is currently known about MNK and its variants, the efficacy and problems of currently accepted treatments and finally therapeutic targets in MNK. PMID- 12540289 TI - Cerebrospinal fluid levels of catechols in patients with neurogenic orthostatic hypotension. AB - In multiple system atrophy (MSA) and pure autonomic failure (PAF), orthostatic hypotension (OH) results from deficient noradrenaline release from sympathetic nerves during standing. Post-mortem findings have indicated loss of central noradrenergic cells in both diseases. The present study sought in vivo neurochemical evidence for central noradrenergic deficiency in patients with OH due to MSA or PAF. A total of 28 patients with OH (18 with MSA; 10 with PAF) had cerebrospinal fluid and blood sampled for levels of noradrenaline and its neuronal metabolite dihydroxyphenylglycol. A control group of 44 subjects included 10 elderly normal volunteers, 10 patients with Alzheimer's disease, 18 patients with dysautonomia (postural tachycardia syndrome or neurocardiogenic syncope) and six patients with MSA in the absence of OH. Patients with OH had lower cerebrospinal fluid concentrations of noradrenaline (0.53+/-0.07 nmol/l) and dihydroxyphenylglycol (6.52+/-0.46 nmol/l) than did control subjects (0.90+/ 0.09 and 9.64+/-0.46 nmol/l respectively; P =0.0001). The MSA+OH group had higher plasma levels of both catechols (noradrenaline, 1.31+/-0.16 nmol/l; dihydroxyphenylglycol, 5.08+/-0.43 nmol/l) than did the PAF group (noradrenaline, 0.38+/-0.08 nmol/l; dihydroxyphenylglycol, 2.53+/-0.30 nmol/l; P <0.001), despite similarly low cerebrospinal fluid levels. Among MSA patients, those with OH had lower cerebrospinal fluid levels of noradrenaline and dihydroxyphenylglycol than those without OH (noradrenaline, 1.71+/-0.64 nmol/l; dihydroxyphenylglycol, 10.41+/-1.77 nmol/l respectively; P =0.006). The findings are consistent with central noradrenergic deficiency in both MSA+OH and PAF. In MSA, central noradrenergic deficiency seems to relate specifically to OH. PMID- 12540290 TI - Increased myocardial perfusion and sympathoadrenal activation during mild core hypothermia in awake humans. AB - Potential mechanisms of cold-induced myocardial ischaemia are sympathetically mediated coronary vasoconstriction and/or catecholamine-induced increases in cardiac work. To examine these parameters, 11 human volunteers were each studied on one day with, and on another day without, beta-adrenoceptor blockade. On each day, warm (37 degrees C) saline (control) and cold (4 degrees C) saline (hypothermia) were given intravenously. Myocardial perfusion was assessed by positron emission tomography using H(2)(15)O, and coronary vascular resistance was calculated. Plasma catecholamines were measured to assess sympathoadrenal activation. The core temperature decreased by 1.0 +/- 0.2 degrees C with the cold saline, and was unchanged with warm saline. Myocardial perfusion increased by 20% (P = 0.01) and the rate-pressure product by 33% (P = 0.0004) with cold saline compared with warm saline. beta-Blockade eliminated these increases. Coronary vascular resistance was similar with warm and cold saline, and was unaffected by beta-blockade. Plasma adrenaline increased by 120% and noradrenaline by 251% during cold saline, but were unchanged during warm saline. In conclusion, core hypothermia triggers beta-adrenoceptor-mediated increased cardiac work, sympathoadrenal activation and increased myocardial perfusion. There is no evidence for hypothermia-induced coronary vasoconstriction. PMID- 12540291 TI - Identification and characterization of a prostate-specific androgen-independent protein-binding site in the probasin promoter. AB - In this study we investigated the combination of transcription factors and proteins binding to the proximal part of the prostate-specific probasin (PB) promoter. Using DNaseI in vitro footprinting, several protected regions were identified on the proximal PB promoter (nucleotides -286 to +28 relative to the transcription start site) when nuclear extracts from LNCaP, a human prostate cancer cell line, were used. Four of the protected areas were observed only when LNCaP nuclear extracts treated with synthetic androgen (10 nM R1881) were used. Two other regions, referred to as FPI and FPII, showed protection regardless of the presence or absence of androgen. When DNaseI footprinting was done using other prostate and non-prostate nuclear extracts, protection of the FPII region was only seen in prostate cell lines. These androgen-independent regions were further tested for tissue and binding specificity using the electrophoretic mobility-shift assay. Eight complexes formed with the FPI probe while four complexes were observed with the FPII probe on incubation with the tested nuclear extracts. Methylation protection assays reveal that prostate cancer cell lines yield slightly different protection patterns for some of the protein complexes formed with non-prostate-derived cell lines, suggesting the presence of prostate enriched or -exclusive proteins. Site-directed mutagenesis of the protected nucleotides within FPII resulted in a significant reduction in expression from the PB promoter. Identification of proteins binding to the FPII region revealed the participation of nuclear factor I (NF-I) or a closely related protein, although other unknown proteins are also involved. Defining the DNA and protein components that dictate prostate-specific expression of the PB promoter in an androgen-independent manner would provide a strong basis for the design and development of a gene therapy for systemic treatment of androgen-independent prostate cancer. PMID- 12540292 TI - Stimulation of glycogen synthesis by heat shock in L6 skeletal-muscle cells: regulatory role of site-specific phosphorylation of glycogen-associated protein phosphatase 1. AB - Recent evidence suggests that glycogen-associated protein phosphatase 1 (PP-1(G)) is essential for basal and exercise-induced glycogen synthesis, which is mediated in part by dephosphorylation and activation of glycogen synthase (GS). In the present study, we examined the potential role of site-specific phosphorylation of PP-1(G) in heat-shock-induced glycogen synthesis. L6 rat skeletal-muscle cells were stably transfected with wild-type PP-1(G) or with PP-1(G) mutants in which site-1 (S1) Ser(48) and site-2 (S2) Ser(67) residues were substituted with Ala. Cells expressing wild-type and PP-1(G) mutants, S1, S2 and S1/S2, were examined for potential alterations in glycogen synthesis after a 60 min heat shock at 45 degrees C, followed by analysis of [(14)C]glucose incorporation into glycogen at 37 degrees C. PP-1(G) S1 mutation caused a 90% increase in glycogen synthesis on heat-shock treatment, whereas the PP-1(G) S2 mutant was not sensitive to heat stress. The S1/S2 double mutant was comparable with wild-type, which showed a 30% increase over basal. Heat-shock-induced glycogen synthesis was accompanied by increased PP-1 and GS activities. The highest activation was observed in S1 mutant. Heat shock also resulted in a rapid and sustained Akt/ glycogen synthase kinase 3 beta (GSK-3 beta) phosphorylation. Wortmannin blocked heat-shock-induced Akt/GSK-3 beta phosphorylation, prevented 2-deoxyglucose uptake and abolished the heat-shock-induced glycogen synthesis. Muscle glycogen levels regulate GS activity and glycogen synthesis and were found to be markedly depleted in S1 mutant on heat-shock treatment, suggesting that PP-1(G) S1 Ser phosphorylation may inhibit glycogen degradation during thermal stimulation, as S1 mutation resulted in excessive glycogen synthesis on heat-shock treatment. In contrast, PP 1(G) S2 Ser phosphorylation may promote glycogen breakdown under stressful conditions. Heat-shock-induced glycogenesis appears to be mediated via phosphoinositide 3-kinase/Akt-dependent GSK-3 beta inactivation as well as phosphoinositide 3-kinase-independent PP-1 activation. PMID- 12540293 TI - Upstream stimulatory factor (USF) and neurogenic differentiation/beta-cell E box transactivator 2 (NeuroD/BETA2) contribute to islet-specific glucose-6 phosphatase catalytic-subunit-related protein (IGRP) gene expression. AB - Islet-specific glucose-6-phosphatase (G6Pase) catalytic-subunit-related protein (IGRP) is a homologue of the catalytic subunit of G6Pase, the enzyme that catalyses the final step of the gluconeogenic pathway. The analysis of IGRP chloramphenicol acetyltransferase (CAT) fusion-gene expression through transient transfection of islet-derived beta TC-3 cells revealed that multiple promoter regions, located between -306 and -97, are required for maximal IGRP-CAT fusion gene expression. These regions correlated with trans -acting factor-binding sites in the IGRP promoter that were identified in beta TC-3 cells in situ using the ligation-mediated PCR (LMPCR) footprinting technique. However, the LMPCR data also revealed additional trans -acting factor-binding sites located between -97 and +1 that overlap two E-box motifs, even though this region by itself conferred minimal fusion-gene expression. The data presented here show that these E-box motifs are important for IGRP promoter activity, but that their action is only manifest in the presence of distal promoter elements. Thus mutation of either E box motif in the context of the -306 to +3 IGRP promoter region reduces fusion gene expression. These two E-box motifs have distinct sequences and preferentially bind NeuroD/BETA2 (neurogenic differentiation/beta-cell E box transactivator 2) and upstream stimulatory factor (USF) in vitro, consistent with the binding of both factors to the IGRP promoter in situ, as determined using the chromatin-immunoprecipitation (ChIP) assay. Based on experiments using mutated IGRP promoter constructs, we propose a model to explain how the ubiquitously expressed USF could contribute to islet-specific IGRP gene expression. PMID- 12540294 TI - Post-genomic insights into plant nodulation symbioses. AB - Several legume genes involved in establishing nitrogen fixation have been discovered using functional genomics; when mutated, the genes affect symbioses, and all encode receptor kinases. This provides long-awaited insights into a complex plant-bacterium interaction and heralds the possibility of extending the range of plants susceptible to nitrogen-fixing nodulation. PMID- 12540295 TI - The genomics of insecticide resistance. AB - Genomic technologies are revealing several mechanisms of insecticide resistance involving enhanced detoxification or reduced target-site sensitivity that had previously defied molecular analyses. Genome projects are also revealing some potentially far-reaching consequences for pest-insect genomes of the rapid accumulation of multiple resistance mutations in very short periods of evolutionary time. PMID- 12540296 TI - The sigma70 family of sigma factors. AB - Members of the sigma70 family of sigma factors are components of the RNA polymerase holoenzyme that direct bacterial or plastid core RNA polymerase to specific promoter elements that are situated 10 and 35 base-pairs upstream of transcription-initiation points. Members of the sigma70 family also function as contact points for some activator proteins, such as PhoB and lambda(cl), and play a role in the initiation process itself. The primary sigma factor, which is essential for general transcription in exponentially growing cells, is reversibly associated with RNA polymerase and can be replaced by alternative sigma factors that co-ordinately express genes involved in diverse functions, such as stress responses, morphological development and iron uptake. On the basis of gene structure and function, members of the sigma70 family can broadly be divided into four main groups. Sequence alignments of the sigma70 family members reveal that they have four conserved regions, although the highest conservation is found in regions 2 and 4, which are involved in binding to RNA polymerase, recognizing promoters and separating DNA strands (so-called 'DNA melting'). The division of the linear sequence of sigma70 factors into four regions is largely supported by recent structural data indicating that primary sigma factors have three stable domains that incorporate regions 2, 3 and 4. Furthermore, structures of the RNA polymerase holoenzyme have revealed that these domains of sigma70 are spread out across one face of RNA polymerase. These structural data are starting to illuminate the mechanistic role of sigma factors in transcription initiation. PMID- 12540297 TI - Assessing unmodified 70-mer oligonucleotide probe performance on glass-slide microarrays. AB - BACKGROUND: Long oligonucleotide microarrays are potentially more cost- and management-efficient than cDNA microarrays, but there is little information on the relative performance of these two probe types. The feasibility of using unmodified oligonucleotides to accurately measure changes in gene expression is also unclear. RESULTS: Unmodified sense and antisense 70-mer oligonucleotides representing 75 known rat genes and 10 Arabidopsis control genes were synthesized, printed and UV cross-linked onto glass slides. Printed alongside were PCR-amplified cDNA clones corresponding to the same genes, enabling us to compare the two probe types simultaneously. Our study was designed to evaluate the mRNA profiles of heart and brain, along with Arabidopsis cRNA spiked into the labeling reaction at different relative copy number. Hybridization signal intensity did not correlate with probe type but depended on the extent of UV irradiation. To determine the effect of oligonucleotide concentration on hybridization signal, 70-mers were serially diluted. No significant change in gene-expression ratio or loss in hybridization signal was detected, even at the lowest concentration tested (6.25 microm). In many instances, signal intensity actually increased with decreasing concentration. The correlation coefficient between oligonucleotide and cDNA probes for identifying differentially expressed genes was 0.80, with an average coefficient of variation of 13.4%. Approximately 8% of the genes showed discordant results with the two probe types, and in each case the cDNA results were more accurate, as determined by real-time PCR. CONCLUSIONS: Microarrays of UV cross-linked unmodified oligonucleotides provided sensitive and specific measurements for most of the genes studied. PMID- 12540298 TI - Towards reconstruction of gene networks from expression data by supervised learning. AB - BACKGROUND: Microarray experiments are generating datasets that can help in reconstructing gene networks. One of the most important problems in network reconstruction is finding, for each gene in the network, which genes can affect it and how. We use a supervised learning approach to address this question by building decision-tree-related classifiers, which predict gene expression from the expression data of other genes. RESULTS: We present algorithms that work for continuous expression levels and do not require a priori discretization. We apply our method to publicly available data for the budding yeast cell cycle. The obtained classifiers can be presented as simple rules defining gene interrelations. In most cases the extracted rules confirm the existing knowledge about cell-cycle gene expression, while hitherto unknown relationships can be treated as new hypotheses. CONCLUSIONS: All the relations between the considered genes are consistent with the facts reported in the literature. This indicates that the approach presented here is valid and that the resulting rules can be used as elements for building and explaining gene networks. PMID- 12540299 TI - MAPPFinder: using Gene Ontology and GenMAPP to create a global gene-expression profile from microarray data. AB - MAPPFinder is a tool that creates a global gene-expression profile across all areas of biology by integrating the annotations of the Gene Ontology (GO) Project with the free software package GenMAPP http://www.GenMAPP.org. The results are displayed in a searchable browser, allowing the user to rapidly identify GO terms with over-represented numbers of gene-expression changes. Clicking on GO terms generates GenMAPP graphical files where gene relationships can be explored, annotated, and files can be freely exchanged. PMID- 12540301 TI - Children with fever and vomiting benefit from immediate antibiotics for acute otitis media. AB - Starting antibiotics immediately in children with acute otitis media accompanied by fever or vomiting results in better symptom relief and decreased sleep disturbance when measured after 3 days, as compared with no treatment. Parents who consider these outcomes important may prefer not to delay antibiotic treatment. Conversely, children without fever or vomiting tend to have the same duration of symptoms regardless of antibiotic treatment and are suitable for a "wait and see" approach. PMID- 12540302 TI - Six-item screening tool is sensitive for dementia. AB - This study provides excellent evidence that a 6-item screening tool based on orientation to date and a 3-item short-term recall is sensitive for dementia. Clinicians should consider using this simple screen in an outpatient setting, keeping in mind that the final diagnosis of dementia is a clinical judgment after full assessment and that this screen was not intended for use in following patients over time. Caution also should be exercised in extending these results to patients in the hospital, who were not included in this trial. PMID- 12540303 TI - Should patients with coronary disease and high homocysteine take folic acid? AB - All patients with known coronary artery disease should take prescription strength (1 mg/d) folic acid, vitamin B12 (400 microg/d), and vitamin B6 (10 mg/d), which have few if any known adverse effects. In this study, therapy to reduce homocysteine levels with prescription strength folic acid (1 mg) and vitamins B12 and B6 for 6 months following coronary angioplasty reduced the risk of need for revascularization of target lesions and of overall adverse cardiac events at least 6 months following cessation of therapy. Based on this study, it is unknown whether the benefit is related to baseline homocysteine levels or whether there is further benefit to continuing treatment beyond 6 months. Over-the-counter folic acid supplements (800 microg or less) were not studied and may not be as beneficial. PMID- 12540304 TI - Watchful waiting is reasonable for gallstone symptoms. AB - Not all patients with symptomatic cholelithiasis require surgery. Nearly half of patients with symptomatic but uncomplicated gallstone disease can be managed successfully with observation and minor dietary changes. This option is a safe one we can offer our patients. PMID- 12540305 TI - Korean red ginseng effective for treatment of erectile dysfunction. AB - Korean red ginseng (Panax ginseng) is a safe, widely available alternative remedy that improves patients' ability to achieve and maintain an erection sufficient for intercourse, even in a population with severe erectile dysfunction. It is a reasonable, nonprescription treatment, especially for men with reservations about taking sildenafil (Viagra). A 500-mg capsule of Korean red ginseng costs about 6 cents, compared with $10 for a tablet of sildenafil. PMID- 12540306 TI - Metronidazole gel ineffective for minimally abnormal Pap. AB - Empiric treatment of women with minimally abnormal Papanicolaou smears (limited by inflammation, benign, or reactive cellular changes) with 0.75% metronidazole vaginal gel is ineffective in yielding a higher rate of reversion to normal cytology when compared with no treatment. PMID- 12540307 TI - Early radical prostatectomy improves disease-specific but not overall survival. AB - For now, a reasonable strategy is to consider watchful waiting as an acceptable alternative to radical prostatectomy for patients with early prostate cancer and a lifespan of less than 10 years. For other patients, discuss the benefits and risks of the treatment options, balancing expected side effects of the operation and the impact of other illnesses on survival with the possible benefit of the operation or other kinds of treatment. We still lack sufficient evidence whether early detection by PSA screening can reduce morbidity or mortality. In this study, radical prostatectomy for early prostate cancer decreased disease-specific mortality, but did not improve overall mortality. A companion study (1) showed that non-nerve-sparing radical prostatectomy yielded no difference in subjective quality of life, although clinically important increases in erectile dysfunction (number needed to harm [NNH]=3) and urinary leakage (NNH=4) did occur, compared with watchful waiting. Clinicians should understand that these results might not apply to patients with highly undifferentiated cancer; patients identified by screening to have elevated prostate-specific antigen (PSA) concentrations and no clinically symptomatic disease; or patients with significant comorbidities. PMID- 12540308 TI - Suturing unnecessary for hand lacerations under 2 cm. AB - Hand lacerations less than 2 cm long without tendon, joint, fracture, or nerve complications and not involving the nail bed can be cleaned and dressed without suturing, with similar cosmetic results and time to resume normal activities. Moreover, managing these uncomplicated hand lacerations conservatively could result in better use of medical resources and improved patient satisfaction due to less pain and less time spent in the emergency department. PMID- 12540309 TI - Early invasive strategy for acute cardiac ischemia is cost effective. AB - In patients with unstable angina and non-ST segment myocardial infarction treated with aspirin, heparin, and tirofiban, an early invasive strategy with routine angiography and appropriate revascularization has better clinical outcomes, at a relatively minimal increase in cost. PMID- 12540310 TI - Warfarin plus aspirin more effective than aspirin alone for secondary prevention of MI. AB - Compared with aspirin alone, aspirin plus warfarin (goal for international normalized ratio, 2-2.5) or warfarin alone (goal for international normalized ratio, 2.8-4.3) results in fewer re-infarctions and thromboembolic events. Treating 1000 patients for 1 year would result in approximately 10 fewer reinfarctions and 3 fewer strokes at a cost of 4 more major bleeding episodes. In addition, many patients will not be able to tolerate warfarin therapy. For highly motivated patients at low risk of bleeding, warfarin or warfarin plus aspirin is more effective than aspirin for secondary prevention of myocardial infarction. PMID- 12540311 TI - Effective management of obesity. AB - Successful treatment of obesity usually requires multiple interventions. The choice of therapies should be guided by the initial assessment of a patient's degree of obesity and comorbid conditions, if present. A variety of interventions can achieve short-term weight loss, but rebound weight gain is common when therapy is stopped. Thus, programs for weight maintenance are critical to ultimate success. PMID- 12540312 TI - Management of acne. AB - Precise classification methods are used to define acne according to type (comedonal, papulopustular, or nodular) and severity. The relative effectiveness of several topical and systemic agents has been established in clinical trials, making possible an algorithm of specific treatment decisions based on acne classification. PMID- 12540313 TI - A swollen knee. PMID- 12540314 TI - Screening for handicapping hearing loss in the elderly. AB - OBJECTIVE: To compare 2 screening methods for unrecognized handicapping hearing loss in the elderly. STUDY DESIGN: Cross-sectional study. POPULATION: Five hundred forty-six older individuals who underwent audiometry at biennial examination 22 of the Framingham Heart Study and who took the Hearing Handicap Inventory for the Elderly-Screening (HHIE-S) questionnaire. OUTCOMES MEASURED: The 2 screening methods were the 10-item HHIE-S and 1 global question: "Do you have a hearing problem now?" The gold standard was an audiogram showing a pure tone threshold of 40 dB HL or higher at 1 and 2 kHz in one ear or at 1 or 2 kHz in both ears. Both screening methods were compared with the gold standard in terms of sensitivity, specificity, and predictive values. The 10-item screening version of the HHIE-S (cutoff score between 8 and 10) had a sensitivity of 35% and a specificity of 94% for detecting the criterion hearing loss. The global subjective measure had greater sensitivity (71%) but lower specificity (71%) than the HHIE-S. Combining the global question and the HHIE-S items failed to improve the specificity of the global question or the sensitivity of the HHIE-S. CONCLUSIONS: The global measure of hearing loss was more effective than the detailed questionnaire in identifying older individuals with unrecognized handicapping hearing loss. Primary care physicians are encouraged to ask their patients whether they have a hearing problem and refer patients who do for formal hearing testing. PMID- 12540315 TI - Clinical inquiries. How accurate is the clinical diagnosis of pneumonia? PMID- 12540316 TI - Clinical inquiries. Who should have colposcopy? PMID- 12540317 TI - Clinical inquiries. What is the best treatment for bronchiolitis? PMID- 12540318 TI - Clinical inquiries. Does surgery for carpal tunnel syndrome improve outcomes? PMID- 12540319 TI - Nortriptyline effective for smoking cessation. AB - Nortriptyline (Pamelor), in combination with weekly behavioral therapy, is effective in helping highly motivated smokers to quit. The medication may be an alternative for patients who cannot tolerate or do not benefit from bupropion. Given the high motivation of the group and the extensive behavioral therapy they also received, results are not likely to be as good in typical practice. PMID- 12540320 TI - Revascularization not superior to conservative treatment of acute coronary syndromes. AB - Conservative treatment was nearly as effective as immediate catheterization and surgical intervention in patients presenting with acute coronary syndrome. No difference was noted in the risk of death or myocardial infarction in either group. Patients were less likely to experience refractory angina when evaluated at 4 months and after 1 year when treated aggressively (numbers needed to treat [NNT]=20). Saving 1 readmission for refractory angina at the cost of performing 19 interventions that have no effect on the patient may not be reasonable. PMID- 12540321 TI - Lansoprazole ineffective for functional dyspepsia. AB - Patients with functional dyspepsia, which involves no organic disease and no reflux, do not benefit from lansoprazole, a proton pump inhibitor. Any improvement seen is probably related to a placebo effect. Prescribing proton pump inhibitors for these patients just for their placebo effect is going to further raise health care costs. Antacids and H2 blockers are less expensive alternatives. PMID- 12540322 TI - Benefits of tamoxifen for breast cancer prevention do not always outweigh overall risks. AB - In women at high risk of breast cancer, tamoxifen is effective in reducing the incidence of the disease. The intervention, however, is associated with significant risk. During the 5-year period of this study, although the number of breast cancers was reduced, the number of serious adverse effects and deaths were higher in the treated group. Women at a lower risk of breast cancer than those studied would be even less likely to benefit from tamoxifen while risking the same serious adverse outcomes. For the small proportion of women with a high risk of breast cancer and a low risk of adverse events, discussion of this intervention may be warranted. PMID- 12540323 TI - Shock wave therapy ineffective for plantar fasciitis. AB - Extracorporeal shock wave therapy (ESWT) cannot be recommended to improve pain and function in patients with plantar fasciitis based on the results of this study. Although previous studies do report a benefit from ESWT, this study appears to represent a higher level of evidence than was previously available for evaluating the efficacy of this therapy. An updated meta-analysis combining all the studies on ESWT will be useful. PMID- 12540324 TI - Stages of change analysis of smokers attending clinics for the medically underserved. AB - OBJECTIVE: To determine whether smokers at clinics providing care for the medically underserved can be characterized according to the transtheoretical stages of change model. STUDY DESIGN: Prospective, descriptive study. POPULATION: Smokers in the waiting rooms of clinics providing care for the medically underserved. OUTCOMES MEASURED: Standardized questionnaires that assessed stages of change, processes of change, decisional balance, and self-efficacy and temptation. RESULTS: The smoking rate of subjects interviewed at 4 clinics was 44%. Two hundred current smokers completed the questionnaires. Smokers claiming that they planned to quit within 6 months scored higher on experiential process statements that are consistent with quitting smoking than did smokers who claimed they were not planning to quit within 6 months. They also scored higher on behavioral statements related to quitting. Concerns about the negative aspects of smoking were more important to smokers planning to quit than to smokers not planning to quit, whereas the statements assessing positive aspects of smoking were rated the same. Fifty-five percent of the smokers were smoking a pack or more each day and reported smoking more during negative situations and from habit than did smokers who smoked less than a pack a day. CONCLUSIONS: Smokers planning to quit who still smoke at least a pack a day may benefit from counseling to decrease smoking for specific reasons or from pharmacologic aids. Smokers at the clinics who planned to quit smoking reported experiences and behaviors that were consistent with their stated desire to quit and should be counseled in the same fashion as smokers from more traditional practices. PMID- 12540325 TI - Effectiveness of chart prompt about immunizations in an urban health center. AB - OBJECTIVE: To determine whether a nurse-initiated chart review and prompt to physicians is an effective method to increase immunization rates. STUDY DESIGN: This study was a controlled trial with systematic assignment of children to intervention or control groups based on chart number. Each day, a nurse reviewed the charts of children to be seen that day who were in the intervention group. The nurse prepared a 1-page form about the child's immunization status that requested permission from the physician to administer needed vaccines and attached the form to the chart. The duration of the study period was 1 year. POPULATION: Nine hundred ninety-seven pediatric patients attending 2 inner-city primary care health centers. OUTCOME MEASURED: On-time immunization rates in both groups. RESULTS: Among children eligible to receive vaccines during the study period, a higher percentage in the intervention group received on-time vaccines for diphtheria/tetanus/pertussis-4 (DTP4; 51% vs 36%; P=.03), oral polio vaccine 3 (OPV3; 70% vs 56%, P=.04), and measles/mumps/rubella-1 (42% vs 26%; P=.01) than did children in the control group. No statistically significant differences were noted for DTP3, DTP5, hepatitis B3, or OPV4. No statistically significant difference was noted for the combined series (ie, all age-appropriate immunizations as recommended by the 1995 Childhood Immunization Schedule of the Centers for Disease Control and Prevention). CONCLUSIONS: The chart prompt increased on-time immunizations for some antigens. PMID- 12540327 TI - The hospitalist movement and family practice-an uneasy fit. PMID- 12540326 TI - Primary care family physicians and 2 hospitalist models: comparison of outcomes, processes, and costs. AB - OBJECTIVES: To compare the care provided by family practice primary care physicians with that provided by 2 hospitalist models: critical care hospitalists and rotating residency faculty family physician hospitalists. STUDY DESIGN: Retrospective chart review. A health maintenance organization mandated that all patients be admitted to a critical care hospitalist team. The family physician hospitalists admitted all other residency patients and patients of some community family physicians. The primary care physicians admitted all their other patients. We adjusted for disease severity by using the Pneumonia Severity Index, age, sex, and comorbidities. POPULATION: Adults admitted with pneumonia to our private urban community hospital. Exclusions included patients with nosocomial pneumonia, human immunodeficiency virus, and acquired immunodeficiency syndrome. OUTCOMES MEASURED: Primary (adjusted for age, sex, comorbidities, and disease severity): hospital charges, length of stay, in-hospital mortality, readmissions, and returns to the emergency room. Secondary: chest radiographs, intensive care use, blood and sputum cultures, compliance with American Thoracic Society guidelines, lifestyle and end-of-life counseling. RESULTS: Of 97 patients, 21 were admitted to the critical care hospitalists, 53 to the family physician hospitalists, and 23 to primary care physicians. The mean charge ($5680) by the primary care physicians was significantly lower than that of the critical care hospitalists ($10,231; P=.005) and trended toward being lower than that of the family physician hospitalists ($7699; P=.08). The patients of critical care and family physician hospitalists had longer mean lengths of stay (critical care hospitalists, 3.8 days; family physician hospitalists, 3.9 days) than did those of the primary care physicians (2.6 days; P=.04 and.01, respectively). Compared with the primary care physicians, the critical care hospitalists were more likely to obtain at least 2 chest x-rays (odds ratio, 4.1; 95% confidence interval, 1.1 15.5) and trended toward increased odds of lengthy stay in the intensive care unit (odd ratio, 2.9; 95% confidence interval, 0.6-14.6). We found no other significant differences in primary or secondary outcomes. CONCLUSIONS: Claims of better and cheaper care by hospitalists need further investigation. Meanwhile, the use of hospitalists should not be mandated, and the use of family physicians as hospitalists should be considered a good alternative to the use of subspecialists. PMID- 12540328 TI - Urea breath testing and analysis in the primary care office. AB - The 13C-urea breath test provides accurate, noninvasive diagnosis of active Helicobacter pylori infection and can document posttherapy cure. This study evaluated point-of-care testing with onsite sample analysis with the use of a desktop infrared spectrophotometer. Ambulatory patients (N=320) underwent 13C urea breath testing, and breath samples were analyzed immediately by clinic staff with no prior breath testing experience. Duplicate samples were sent to a reference laboratory, and the results of both methods were compared. Point-of care testing was simple, with an overall agreement of 99.1%. Accurate near patient 13C-urea breath testing is now practical in the primary care setting even when done by inexperienced personnel. PMID- 12540329 TI - Patient and physician explanatory models for acute bronchitis. AB - OBJECTIVES: Our goals were to develop explanatory models to better understand how physicians diagnose and treat acute bronchitis; to describe patient expectations and needs when experiencing an episode of acute bronchitis; and to enhance communication between physician and patient. STUDY DESIGN: We used qualitative, semi-structured, in-depth interviews to generate patient and physician explanatory models. POPULATION: We had a purposeful, homogeneous sample of 30 family physicians and 30 adult patients. OUTCOMES MEASURED: Our multidisciplinary team of investigators used an editing style of analysis to develop patient and physician explanatory models based on the following topics: (1) what caused my illness/etiology, (2) what symptoms I had/onset of symptoms, (3) what my sickness did to me/pathophysiology, (4) how severe is my sickness/course of illness, and (5) what kind of treatment should I receive/treatment. RESULTS: We found that patient and physician models were congruous for symptoms of acute bronchitis and incongruous for etiology and course of illness. Models were congruous for treatment, although for different reasons. CONCLUSIONS: Patients may have a very vague understanding of the process of infection and the difference between bacteria and viruses. Compounding this confusion is frequent miscommunication from physicians regarding the clinical course of untreated illness. These factors and non-communicated expectations from patients and fear of missing something on the part of physicians contribute to the decision to treat with antibiotics. PMID- 12540331 TI - Efficacy of daily hypertonic saline nasal irrigation among patients with sinusitis: a randomized controlled trial. AB - OBJECTIVES: To test whether daily hypertonic saline nasal irrigation improves sinus symptoms and quality of life and decreases medication use in adult subjects with a history of sinusitis. STUDY DESIGN: Randomized controlled trial. Experimental subjects used nasal irrigation daily for 6 months. POPULATION: Seventy-six subjects from primary care (n=70) and otolaryngology (n=6) clinics with histories of frequent sinusitis were randomized to experimental (n=52) and control (n=24) groups. OUTCOMES MEASURED: Primary outcome measures included the Medical Outcomes Survey Short Form (SF-12), the Rhinosinusitis Disability Index (RSDI), and a Single-Item Sinus-Symptom Severity Assessment (SIA); all 3 were completed at baseline, 1.5, 3, and 6 months. Secondary outcomes included daily assessment of compliance and biweekly assessment of symptoms and medication use. At 6 months, subjects reported on side effects, satisfaction with nasal irrigation, and the percentage of change in their sinus-related quality of life. RESULTS: No significant baseline differences existed between the 2 groups. Sixty nine subjects (90.8%) completed the study. Compliance averaged 87%. Experimental group RSDI scores improved from 58.4 -/+ 2.0 to 72.8 -/+ 2.2 (P < or =.05) compared with those of the control group (from 59.6 -/+ 3.0 to 60.4 -/+ 1.1); experimental group SIA scores improved from 3.9 -/+ 0.1 to 2.4 -/+ 0.1 (P < or =.05) compared with those of the control group (from 4.08 -/+ 0.15 to 4.07 -/+ 0.27). The number needed to treat to achieve 10% improvement on RSDI at 6 months was 2.0. Experimental subjects reported fewer 2-week periods with sinus-related symptoms (P <.05), used less antibiotics (P <.05), and used less nasal spray (P =.06). On the exit questionnaire 93% of experimental subjects reported overall improvement of sinus-related quality of life, and none reported worsening (P <.001); on average, experimental subjects reported 57 -/+ 4.5% improvement. Side effects were minor and infrequent. Satisfaction was high. We found no statistically significant improvement on the SF-12. CONCLUSIONS: Daily hypertonic saline nasal irrigation improves sinus-related quality of life, decreases symptoms, and decreases medication use in patients with frequent sinusitis. Primary care physicians can feel comfortable recommending this therapy. PMID- 12540330 TI - Managing musculoskeletal complaints with rehabilitation therapy: summary of the Philadelphia Panel evidence-based clinical practice guidelines on musculoskeletal rehabilitation interventions. AB - OBJECTIVE: The Philadelphia Panel recently formulated evidence-based guidelines for selected rehabilitation interventions in the management of low back, knee, neck, and shoulder pain. STUDY DESIGN: The guidelines were developed with the use of a 5-step process: define the intervention, collect evidence, synthesize results, make recommendations based on the research, and grade the strength of the recommendations. POPULATION: Outpatient adults with low back, knee, neck, or shoulder pain without vertebral disk involvement, scoliosis, cancer, or pulmonary, neurologic, cardiac, dermatologic, or psychiatric conditions were included in the review. OUTCOMES MEASURED: To prepare the data, systematic reviews were performed for low back, knee, neck, and shoulder pain. Therapeutic exercise, massage, transcutaneous electrical nerve stimulation, thermotherapy, ultrasound, electrical stimulation, and combinations of these therapies were included in the literature search. Studies were identified and analyzed based on study type, clinical significance, and statistical significance. CONCLUSIONS: The Philadelphia Panel guidelines recommend continued normal activity for acute, uncomplicated low back pain and therapeutic exercise for chronic, subacute, and postsurgical low back pain; transcutaneous electrical nerve stimulation and exercise for knee osteoarthritis; proprioceptive and therapeutic exercise for chronic neck pain; and the use of therapeutic ultrasound in the treatment of calcific tendonitis of the shoulder. PMID- 12540332 TI - Cancer recurrence and mortality in women using hormone replacement therapy: meta analysis. AB - OBJECTIVES: We compared the risk of cancer recurrence and all-cause mortality among users and nonusers of estrogen replacement therapy (ERT) after the diagnosis of breast cancer. STUDY DESIGN: This was a systematic review of original research. Eligible studies were reviewed by 2 investigators who independently extracted data from each study according to a predetermined form and assessed each study for validity on standard characteristics. Meta-analyses were performed with Review Manager 4.1 to provide a summary of relative risks of cancer recurrence and mortality. POPULATION: Studies included 717 subjects who used hormone replacement therapy (HRT) at some time after their diagnosis of breast cancer, as well as 2545 subjects who did not use HRT. OUTCOMES MEASURED: Outcomes included breast cancer recurrence and all-cause mortality. RESULTS: Nine independent cohort studies and one 6-month pilot randomized controlled trial were identified. Studies were of variable quality. Breast cancer survivors using ERT experienced no increase in the risk of recurrence compared with controls (relative risk, 0.72; 95% confidence interval, 0.47-1.10) and had significantly fewer deaths (3.0%) than did the nonusers (11.4%) over the combined study periods (relative risk, 0.18; 95% confidence interval, 0.10-0.31). All tests for heterogeneity were nonsignificant. CONCLUSIONS: Although limited by observational design, existing research does not support the universal withholding of ERT from well-informed women with a previous diagnosis of low-stage breast cancer. Long term randomized controlled trials are needed. PMID- 12540333 TI - Improved detection of depression in primary care through severity evaluation. AB - OBJECTIVES: To determine whether the use of a symptom severity measure to augment an existing Diagnostic and Statistical Manual of Mental Disorders-Third Edition, Revised (DSM-III-R) criteria-based depression screener (PRIME-MD) would decrease the difficulties associated with depression screening in primary care by filtering out patients with minimal impairment. STUDY DESIGN: The study design was secondary data analysis. POPULATION: The study sample comprised 1317 patients, with intentional oversampling by ethnicity and sex, presenting for routine care at a university family practice center in Galveston, Texas. OUTCOMES MEASURED: The primary outcomes were cross-sectional, health-related quality-of life outcomes of subjects who met symptom severity criteria as well as criteria for a DSM-III-R mood disorder. Health care utilization outcomes were examined as secondary outcomes. RESULTS: The combination of a 6-item depression severity instrument and the PRIME-MD resulted in 71% of depressed subjects being categorized as severely symptomatic and 29% as minimally symptomatic. Severely symptomatic subjects had significantly worse SF-36 Mental Health Component Summary scale (MCS) scores than did minimally symptomatic subjects (32.8 vs 43.5, P <.05). Minimally symptomatic subjects had MCS scores similar to those of a third group of subjects who did not meet DSM-III-R "threshold" criteria for mood disorder but who were severely symptomatic. Adjusted health care utilization was higher for the initial 3-month charge period in the severely symptomatic depressed subjects compared with minimally symptomatic depressed subjects ($679.20 vs $462.38, P <.05). CONCLUSIONS: The 6-item depression severity measure effectively separated patients meeting DSM-III-R "threshold" depression criteria into 2 groups: one presenting with severe symptoms and impairment and the other presenting with mild symptoms and significantly less impairment. A strategy of initial screening using a brief depression severity instrument, followed with a DSM criteria-based instrument, could decrease the immediate clinician workload by one third and focus treatment on those most likely to benefit. PMID- 12540334 TI - Prevalence of overactive bladder and urinary incontinence. AB - We conducted a cross-sectional study in Italy among men at least 50 years old and women at least 40 years old who consecutively visited their general practitioners. Patients were asked about the frequency of symptoms of overactive bladder and urinary incontinence. A total of 9613 men (mean age, 64.8 years; range, 50-98 years) and 13,365 women (mean age, 60.3 years; range, 40-98 years) were identified by 774 general practitioners. The frequencies of overactive bladder were 3.0% (95% confidence interval, 2.7-3.5) in men and 1.1% (95% confidence interval, 0.9-1.3) in women. The corresponding frequencies for urinary incontinence were 8.3% (95% confidence interval, 7.7-8.9) in men and 10.2% (95% confidence interval, 9.6-10.8) in women. PMID- 12540335 TI - Clinical inquiries. What are effective strategies for reducing the risk of steroid-induced osteoporosis? PMID- 12540336 TI - Clinical inquiries. What are the indications for urodynamic testing in older adults with incontinence? PMID- 12540337 TI - Clinical inquiries. What is the best diagnostic approach to parasthesias of the hand? PMID- 12540339 TI - Spirometry in old age. PMID- 12540340 TI - Explicit evidence-based prescribing criteria--an important step in achieving quality therapeutics in nursing homes. PMID- 12540341 TI - Growing knowledge about hip fracture in older people. PMID- 12540342 TI - Urinary incontinence in nursing homes for older people. AB - At least 50% of nursing home residents in Britain and North America suffer from urinary incontinence. It is associated with resident and staff morbidity. The assessment and management of such residents will depend on the capacity of the care staff and the capability of the resident. The minimum data set and resident assessment protocol may have a role in the assessment of incontinent residents. Behavioural strategies are more likely to be beneficial than drug treatment. PMID- 12540343 TI - Non-pharmacological management of primary and secondary insomnia among older people: review of assessment tools and treatments. AB - BACKGROUND: primary and secondary insomnia, especially among older adults, is frequently encountered by family physicians. Pharmacological interventions, although effective in some circumstances, can be detrimental in others. Non pharmacological management of insomnia may allow the patients to self-administer the treatment. OBJECTIVES: review of published literature of assessment tools and treatments for primary and secondary insomnia. RESULTS: two frequently used self reporting methods for obtaining sleep data are sleep diaries and Pittsburg Sleep Quality Index. A large amount of research supports the use of non-pharmacological treatments such as stimulus control, sleep restriction, sleep hygiene education, cognitive therapy, multi-component therapy and paradoxical intention. CONCLUSION: assessing the nature of insomnia by using an effective assessment tool and providing patients with a non-pharmacological treatment should be the first intervention for insomnia. It is shown that non-pharmacological treatments for primary and secondary insomnia are feasible and effective alternatives to the use of benzodiazepines, and that family physicians should consider these when managing older patients with insomnia. PMID- 12540344 TI - Visual factors should be assessed in older people presenting with falls or hip fracture. AB - Visual impairment, although not routinely assessed, is an important risk factor for falls and hip fracture in older people. Impaired vision is highly prevalent and commonly unreported in the elderly population particularly in women and those living in nursing homes. Measurement of visual functions such as visual acuity, contrast sensitivity and depth perception may identify older people at risk of falls and hip fracture. Visual loss in older people is correctable in most cases. Intervention strategies, for example, change of glasses or cataract extraction may have the potential of improving visual function and preventing falls in older people. PMID- 12540345 TI - A test to assess the mechanical power sustainable during everyday activities in older people. AB - OBJECTIVE: to define with a simple test the power sustainable by older people during everyday occupations. METHODS: nine healthy subjects (65-71 years) performed two series of 5-6 trials on bicycle and arm ergometers at constant power. Throughout the exercise they reported their evaluation of the effort, referred either to perceived whole body exertion or to the working muscles, using Borg's category-ratio, CR10, scale. The exercise was interrupted when CR7 was attained. From a linear regression of the individual data of the work done from the start of each trial to the transition from CR4 to CR5, corresponding to 'somewhat heavy' and 'heavy', and the corresponding duration it was possible to calculate the slope of the work/time relationship. This was considered as the value of power sustainable for leg or arm muscles. RESULTS: on average the sustainable power for the work done on the bicycle ergometer represented 55% of the maximal mechanical power. During the control trial on the bicycle ergometer at a power corresponding to their sustainable power the subjects were able to exercise for 30 min without symptoms of fatigue or discomfort, with an average CR of 3. The average heart rate was 114 bpm and the blood lactate concentration at the end of the exercise was always lower than 4 mmol/l. Sustainable power calculated for the arm muscles was about one quarter that of the leg muscles, but all subjects reported the evaluation of the effort as more difficult. CONCLUSION: the determination of power sustainable for the leg muscles with this simple test appears reliable and accurate. PMID- 12540346 TI - Alpha 1-acid glycoprotein is an independent predictor of in-hospital death in the elderly. AB - OBJECTIVES: to determine the contribution of the two acute phase proteins alpha 1 acid glycoprotein, and C-reactive protein to the prediction of overall mortality in hospitalised elderly patients. DESIGN: prospective cohort study. SETTING: a department of geriatric medicine of Charles Richet Hospital, in the Paris-Ile de France area. SUBJECTS: 433 consecutive patients (mean age 84+/-7 years) admitted for rehabilitation in a department of geriatric medicine. METHODS: clinical and laboratory examinations were performed at baseline. Follow-up ended at hospital discharge or death. Prognostic factors of survival were identified using Cox proportional hazards regression model. RESULTS: compared with the survivor group, the mean serum levels of both C-reactive protein and alpha 1-acid glycoprotein at baseline were higher in the deceased group (44+/-51 mg/l versus 22+/-34 mg/l and 1691+/-69 mg/l versus 1340+/-456 mg/l respectively; P<0.001 for each). Baseline levels of albumin and prealbumin were significantly lower in patients who died than in patients who survived. In multivariate analysis, alpha 1-acid glycoprotein (but not C-reactive protein), previous stroke, previous heart failure, and age emerged as the only parameters significantly and independently related to overall mortality. CONCLUSION: this study shows that the increase in plasma levels at admission of two acute-phase proteins, alpha 1-acid glycoprotein and C-reactive protein, were associated with in-hospital mortality in a population of hospitalised elderly patients. Furthermore, associations of overall mortality with inflammation differed among the markers and only alpha 1-acid glycoprotein entered the multivariate prediction model. Our findings support the hypothesis that alpha 1-acid glycoprotein may be superior to C-reactive protein in mortality risk assessment strategies for elderly patients. PMID- 12540347 TI - Quality of spirometric performance in older people. AB - OBJECTIVES: chronic obstructive pulmonary disease and asthma are major causes of hospitalisation and mortality among older patients but respiratory diseases are often under- or misdiagnosed because spirometry is not extensively used at this age. DESIGN: we examined 715 elderly subjects with respiratory symptoms; all underwent a spirometric test and were administered the Mini Mental State Examination, Activities of Daily Living, Instrumental Activities of Daily Living and Geriatric Depression Scale questionnaires for cognitive, functional and effective evaluation. Their educational level and Body Mass Index were also taken into consideration. RESULTS: a total of 585 patients (81.8%) were able to perform spirometry according to ATS'94 criteria while 130 (18.2%) were unable to do it. As regards educational level, Mini Mental State Examination, Activities of Daily Living and Instrumental Activities of Daily Living scores showed a significant difference (P<0.001) between the groups of patients with high-quality spirometries and those with inadequate ones. There was no difference between the two groups in terms of age, Body Mass Index or Geriatric Depression Scale score. CONCLUSIONS: the majority of elderly subjects can perform spirometry according to international guidelines; age itself cannot be considered a risk factor for a bad spirometric performance, but it becomes influential if it is associated with cognitive and functional impairment. PMID- 12540348 TI - An economic analysis of external hip protector use in ambulatory nursing facility residents. AB - OBJECTIVE: to evaluate the economic impact of external hip protector use in nursing facilities. DESIGN: cost-effectiveness and cost-utility analyses. MEASUREMENTS: event probabilities, intervention effectiveness, treatment costs, and utility of hip fracture state were determined from peer-reviewed medical literature and public use data. METHODS: a societal perspective with an 18-month time horizon was used. A decision tree model was constructed. Expected cost, effectiveness, and incremental cost-effectiveness ratio were calculated. One and two-way sensitivity analyses and threshold analyses were performed. RESULTS: the strategy of using external hip protectors saves approximately US$300 per subject and adds 0.01 quality adjusted life year over 18 months. The model remains dominant over the entire range of input values for all variables except the cost of hip protectors. At the maximum estimated cost of US$695/subject/18 months, the incremental cost-effectiveness ratio was $30,600/quality adjusted life year. The strategy of using hip protectors was cost saving or cost neutral if the cost of hip protector strategy is or=65 years) of normal mental status were eligible for the study. Successful follow-up could be monitored in 120 and 117 patients for 6 and 12 months, respectively. Sixty-nine and 39 patients underwent supervised inpatient rehabilitation in an orthopaedic or geriatric hospital, respectively (intervention groups A and B, respectively) whereas 34 patients received no special rehabilitation as they were directly discharged home (control group C). RESULTS: initially a fall-/surgical-induced reduction (P<0.001) of the main outcome measure (Activities of Daily Living) was observed in all patients. Within 6 months of rehabilitation there was an improvement (P<0.01) in Activities of Daily Living; however the pre-fracture scores were not reached. The same time pattern was seen in group C. Therefore no significant differences between the three groups of patients in approaching the baseline status was visible. Moreover, the one-year total mortality in the studied population with normal mental status averaged 11.7% and did not differ between the three groups. CONCLUSION: based on our measured outcome variables institutional rehabilitation after surgical treatment of hip fracture apparently had no significant impact on mortality and morbidity in older patients of normal mental status. PMID- 12540353 TI - Relationship between changes in depressive symptoms and unhealthy lifestyles in late middle aged and older persons: results from the Longitudinal Aging Study Amsterdam. AB - BACKGROUND: depressed mood is common in late life, more prevalent among the chronically diseased than in the general population, and has various health related consequences. So far, the association between depression and unhealthy lifestyles among chronically diseased has not been examined longitudinally in older persons. PRIMARY OBJECTIVE: to determine if depressed mood is associated with unhealthy lifestyles in late middle aged and older people, with or without chronic somatic diseases. METHODS: in a sample of 1,280 community-dwelling people from the Netherlands, the associations between depressive symptoms and lifestyle domains were analysed cross-sectionally and longitudinally - using logistic regression analyses and multivariate analyses of variance. RESULTS: after controlling for confounders, depressed people (n=176 at baseline) were more likely to be smokers (odds ratio 1.71; 95% confidence interval 1.17-2.52). A persistent depression was associated with an increase in cigarette consumption (P=0.036). Having an emerging depression (n=155) was most likely to co-occur with a person's change from being physically active to being sedentary (relative risk ratio 1.62; 95% confidence interval 1.05-2.52), and was associated with the largest decrease in minutes of physical activity (P=0.038). This effect was not modified or confounded by chronic somatic disease. A persistent depression tended to be associated with incident excessive alcohol use (relative risk-ratio 4.04; 95% confidence interval 0.97-16.09; P=0.056). CONCLUSIONS: depression is associated with smoking behaviour, and with an increase in cigarette consumption. An emerging depression is associated with becoming sedentary, irrespective of a person's disease status at baseline, and is associated with decrease in minutes of physical activity. PMID- 12540354 TI - Depression in older people after fall-related injuries: a prospective study. AB - BACKGROUND: objectives of the study were i) to describe changes in depression in independently living people aged 57 or older with fall-related injuries, and ii) to examine the effect of incomplete recovery of physical functions on depression one year post-injury. METHOD: prospective cohort-study, including a pre-injury baseline and post-injury assessments at 8 weeks, 5 months and one year. The sample consisted of 159 patients who sustained various kinds of fall-related injuries to the limbs. Physical functioning was measured by the Groningen Activity Restriction Scale, depression by the Hospital Anxiety and Depression Scale. Additional variables in the study were age, gender, chronic medical conditions and severity of the injury (three level-groups). Pre- and post-injury levels of depression were compared by using Student's t-test and effect size indices. Hierarchical multivariate regression analysis was used to examine the contribution of change in physical functioning between baseline and one year post injury to depression one year post-injury. RESULTS: severity of the injury was not associated with depression. Mean depression levels of all patients remained stable until 5 months post-injury but increased between 5 months and one year. Physical functioning decreased between baseline and 8 weeks post-injury, increased between 8 weeks and 5 months but did not change after 5 months. One year post-injury, both disability and depression were higher than at baseline. Change in physical functioning between baseline and one year post-injury accounted for 19% of the variance in depression explained by the regression model. CONCLUSIONS: depressive reactions did not occur as long as patients experienced improvement in physical functioning but became manifest as recovery appeared to stagnate. No significant differences in this respect were found between hip fracture patients and patients with other injuries. PMID- 12540355 TI - Infectious aetiologies in elderly patients hospitalised with non-pneumonic lower respiratory tract infection. AB - OBJECTIVE: to identify the infectious aetiologies of non-pneumonic lower respiratory tract infections in hospitalised elderly patients, and to characterise the patients in terms of demographic, clinical and therapeutic variables. DESIGN: a prospective, non-interventional, purely serologically based diagnostic study. SETTING: a tertiary university hospital in southern Israel. SUBJECTS: 133 elderly patients hospitalised for non-pneumonic lower respiratory tract infections. METHODS: paired sera were obtained for each of the hospitalisations and were tested using immunofluorescence or enzyme immunoassay methods to identify 13 different pathogens. Only significant changes in antibody titers or levels between the paired sera were considered diagnostic. RESULTS: at least one infectious aetiology was identified in 77 patients (58%). At least one of seven viral aetiologies was identified in 52 patients (39%). A bacterial aetiology was identified in 27 patients (20%) including Streptococcus pneumoniae in 24 (18%). An atypical bacterium was found in 27 patients (20%) including Mycoplasma pneumoniae in 15 (11%) and Legionella spp. in nine (7%). More than one aetiology was found in 23 patients (17%). One hundred and twenty nine patients (96%) suffered from serious chronic co-morbidity. One hundred and twenty one patients received antibiotics during their hospitalisation, 106 (80%) with a beta lactam and 42 (31%) with another antibiotic. CONCLUSIONS: non-pneumonic lower respiratory tract infection is caused in hospitalised elderly patients by a broad spectrum of aetiological agents, primarily respiratory viruses with a significant, though lesser, prevalence of classical and atypical bacteria. Despite this distribution of aetiologies, most patients are treated with beta lactam antibiotics. The indication for antibiotic therapy in these patients and the choice of antibiotic preparation should be addressed in further studies. PMID- 12540356 TI - Explicit, evidence-based criteria to assess the quality of prescribing to elderly nursing home residents. AB - BACKGROUND: prescribing in nursing homes is frequently suboptimal. Indicators to measure prescribing quality, including appropriateness of prescribing certain drugs or combinations of drugs, to hospital inpatients have been developed previously. OBJECTIVE: to modify prescribing indicators, including appropriateness of prescribing algorithms developed in the hospital setting, for use in nursing homes. DESIGN: an audit of prescribing to patients resident in a random sample of nursing homes on a single day. Setting, subjects: 22 nursing homes in the former South Thames Region selected from lists of nursing homes with more than 35 residents. All residents aged 65 years or over were eligible. METHODS: prescribing indicators, including evidence-based indicators of appropriateness of prescribing benzodiazepines, steroids with beta(2) agonists, antithrombotics with digoxin and aspirin with nitrates were adapted: to reflect where prophylaxis was not justified in terms of quality of life; and for use with primary care clinical records. Indicators were used to evaluate drugs prescribed to each resident to determine whether prescribing was appropriate. RESULTS: 13 indicators were successfully modified and applied. The 934 residents included were prescribed a mean of 5.1 regular items. Only 496/934 (55%) drug sensitivity statements were completed. Although 24% residents received benzodiazepines, clinical data indicated that only 7% received benzodiazepines appropriately. Over three-quarters of residents with ischaemic heart disease received appropriate aspirin therapy, but fewer than half residents with atrial fibrillation received appropriate antithrombotic therapy. It was not possible to derive reference ranges of observed prescribing that included homes demonstrating appropriate prescribing whilst excluding those with inappropriate prescribing. Intra-cluster correlations ranged from 0.027 to 0.335. CONCLUSION: quality of prescribing indicators were successfully modified for the nursing home setting. Application identified suboptimal prescribing to nursing home residents. PMID- 12540357 TI - Hospital re-admissions in relation to acute stroke unit care versus conventional care in elderly patients the first year after stroke: the Goteborg 70+ Stroke study. AB - BACKGROUND: re-hospitalisation after discharge following index stroke varies over time and with age and comorbidity. There is little knowledge about whether stroke unit care reduces the need of re-admissions. OBJECTIVES: to examine whether stroke unit care as compared with care in general medical wards was associated with fewer re-hospitalisations for conditions judged to be secondary to acute stroke and to identify the influence of stroke severity on re-admission rates. DESIGN: we conducted a one-year randomised study to compare the outcome of treatment at an acute stroke unit in a care continuum with the outcome of treatment at general medical wards. SETTINGS: acute and geriatric hospitals in Goteborg, Sweden. SUBJECTS: 216 elderly patients aged >or=70 years discharged to their own homes or to institutionalised living after index stroke. METHODS: comparison of comorbidity classified according to Charlson's morbidity index, re admission rates, length of hospital stay, number of re-admissions and diagnoses between a group treated at a stroke unit and a group treated at general wards. RESULTS: the re-admission rates, length of hospital stay and causes of re admissions did not differ between the two groups. Complications related to the damage to the brain and concomitant heart disease were the most common causes of re-admissions in both groups. Index stroke severity did not influence the re admission rates. CONCLUSIONS: re-admissions for conditions judged to be secondary to acute stroke were equal in the two groups in this prospective study. PMID- 12540358 TI - Hyperkalaemic paralysis. AB - BACKGROUND: hyperkalaemic paralysis is a rare, treatable, and potentially fatal entity presenting with flaccid motor weakness. CASE REPORT: a patient with acute hyperkalaemic paralysis caused by erroneous oral potassium supplementation in the context of chronic renal failure. The clinical picture of quadriplegia was initially interpreted as spinal cord injury due to a pathological cervical fracture. DISCUSSION: the diagnosis of hyperkalaemic paralysis requires a high index of suspicion. Instructions for over-the-counter supplements must be clear and concise to prevent life-threatening medication errors. PMID- 12540359 TI - Anaemia--a common co-existing ailment that is commonly neglected. PMID- 12540360 TI - Infective endocarditis in older people. PMID- 12540361 TI - Palliative care is an important aspect of heart failure management. PMID- 12540362 TI - Evaluation of skills and knowledge on orthostatic blood pressure measurement in elderly patients. PMID- 12540363 TI - Identification of c-kit-positive cells in the mouse ureter: the interstitial cells of Cajal of the urinary tract. AB - The existence of a pacemaker system in the urinary tract capable of orchestrating the movement of filtrated urine from the ureteral pelvis to the distal ureter and lower urinary tract seems intuitive. The coordinated activity necessary for such movement or "peristalsis" would likely require an intricate network of cells with pacemaker-like activity, as is the case with the interstitial cells of Cajal (ICC) of the gut. We investigated whether these putative pacemaker cells of the urinary tract are antigenically similar to ICC of the gut by using immunofluorescence staining for c-kit, a cell-surface marker specific for ICC. Ureteral, urinary bladder, and urethral tissues were harvested from female mice of the WBB6F1 strain, and fixed sections were prepared and stained for c-kit. Cell networks composed of stellate-appearing, c-kit-positive, ICC-like cells were found in the lamina propria and at the interface of the inner longitudinal and outer circular muscle layers of the ureteral pelvis but not in the urinary bladder or urethra. Thus, like in the gut, c-kit-positive, ICC-like cells are present in the urinary tract but appear to be restricted to the proximal ureter of this murine species. PMID- 12540364 TI - Regulation by glucocorticoids and osmolality of expression of ROMK (Kir 1.1), the apical K channel of thick ascending limb. AB - Mechanisms of regulation of ROMK channel mRNA and protein expression in medullary thick ascending limb (MTAL) were assessed in rat MTAL fragments incubated for 7 h. ROMK mRNA was quantified by quantitative RT-PCR and ROMK protein by immunoblotting analysis of crude membranes. Medium hyperosmolality (450 mosmol/kgH(2)O; NaCl plus urea added to isoosmotic medium) increased ROMK mRNA (P < 0.04) and protein (P < 0.006), and 10 nM dexamethasone also increased ROMK mRNA (P < 0.02). Hyperosmolality and dexamethasone had no additive effects on ROMK mRNA. NaCl alone, but not urea or mannitol, reproduced the hyperosmolality effect on ROMK mRNA. 1-Deamino-(8-d-arginine) vasopressin (1 nM) or 0.5 mM 8-bromo-cAMP had no effect per se on ROMK mRNA and protein. However, 8-bromo-cAMP abolished the stimulatory effect of dexamethasone on ROMK mRNA in the isoosmotic but not in the hyperosmotic medium (P < 0.004). In in vivo studies, the abundance of ROMK protein and mRNA increased in adrenalectomized (ADX) rats infused with dexamethasone compared with ADX rats (P < 0.02). These results establish glucocorticoids and medium NaCl concentration as direct regulators of MTAL ROMK mRNA and protein expression, which may be modulated by cAMP-dependent factors. PMID- 12540365 TI - Targeted degradation of ENaC in response to PKC activation of the ERK1/2 cascade. AB - Renal A6 epithelial cells were used to determine the mechanism by which protein kinase C (PKC) decreases epithelial Na(+) channel (ENaC) activity. Activation of PKC reduced relative Na(+) reabsorption to <20% within 60 min. This decrease was sustained over the next 24-48 h. In response to PKC signaling, alpha-, beta-, and gamma-ENaC levels were 0.97, 0.36, and 0.39, respectively, after 24 h, with the levels of the latter two subunits being significantly decreased. The PKC-mediated decreases in beta- and gamma-ENaC were significantly reversed by simultaneous addition of the mitogen-activated protein kinase (MAPK)/extracellular signal regulated kinase-1/2 inhibitors U-0126 and PD-98059. These inhibitors, in addition, protected Na(+) reabsorption from PKC, demonstrating that the MAPK1/2 cascade, in some instances, plays a central role in downregulation of ENaC activity. The effects of PKC on beta- and gamma-ENaC levels were additive with those of inhibitors of transcription (actinomycin D) and translation (emetine and cycloheximide), suggesting that PKC promotes subunit degradation and does not affect subunit synthesis. The bulk of whole cell gamma-ENaC was degraded within 1 h after treatment with inhibitors of synthesis; however, a significant pool was "protected" from inhibitors for up to 12 h. PKC affected this protected pool of gamma-ENaC. Moreover, proteosome inhibitors (MG-132 and lactacystin) reversed PKC effects on this protected pool of gamma-ENaC. Thus PKC signaling via MAPK1/2 cascade activation in A6 cells promotes degradation of gamma-ENaC. PMID- 12540366 TI - Development and characterization of secretin-stimulated secretion of cultured rat cholangiocytes. AB - We sought to develop a cholangiocyte cell culture system that has preservation of receptors, transporters, and channels involved in secretin-induced secretion. Isolated bile duct fragments, obtained by enzyme perfusion of normal rat liver, were seeded on collagen and maintained in culture up to 18 wk. Cholangiocyte purity was assessed by staining for gamma-glutamyl transpeptidase (gamma-GT) and cytokeratin-19 (CK-19). We determined gene expression for secretin receptor (SR), cystic fibrosis transmembrane conductance regulator, Cl(-)/HCO(3)(-) exchanger, secretin-stimulated cAMP synthesis, Cl(-)/HCO(3) exchanger activity, secretin stimulated Cl(-) efflux, and apical membrane-directed secretion in polarized cells grown on tissue culture inserts. Cultured cholangiocytes were all gamma-GT and CK-19 positive. The cells expressed SR and Cl(-)/HCO(3)(-) exchanger, and secretin-stimulated cAMP synthesis, Cl(-)/HCO(3)(-) exchanger activity, and Cl(-) efflux were similar to freshly isolated cholangiocytes. Forskolin (10(-4) M) induced fluid accumulation in the apical chamber of tissue culture inserts. In conclusion, we have developed a novel cholangiocyte line that has persistent HCO(3)(-), Cl(-), and fluid transport functions. This cell system should be useful to investigators who study cholangiocyte secretion. PMID- 12540367 TI - Distribution of interstitial cells of Cajal in tunica muscularis of the canine rectoanal region. AB - Electrical and mechanical activity of the circular muscle layer in the rectoanal region of the gastrointestinal tract undergoes considerable changes in the site of dominant pacemaking activity, frequency, and waveform shape. The present study was performed to determine whether changes in the structural organization of the circular layer or in the density, distribution, and ultrastructure of interstitial cells of Cajal (ICC) could account for this heterogeneity in electrical and mechanical activities. Light microscopy revealed that the structural organization of the circular muscle layer underwent dramatic morphological changes, from a tightly packed layer with poorly defined septa in the proximal rectum to one of discrete muscle bundles separated by large septae in the internal anal sphincter. Kit immunohistochemistry revealed a dense network of ICC along the submucosal and myenteric borders in the rectum, whereas in the internal anal sphincter, ICC were located along the periphery of muscle bundles within the circular layer. Changes in electrical activity within the circular muscle layer can be partially explained by changes in the structure of the muscle layer and changes in the distribution of ICC in the rectoanal region of the gastrointestinal tract. PMID- 12540368 TI - Spatial localization and properties of pacemaker potentials in the canine rectoanal region. AB - The present study investigated the spatial organization of electrical activity in the canine rectoanal region and its relationship to motility patterns. Contraction and resting membrane potential (E(m)) were measured from strips of circular muscle isolated 0.5-8 cm from the anal verge. Rapid frequency [25 cycles/min (cpm)] E(m) oscillations (MPOs, 12 mV amplitude) were present across the thickness of the internal anal sphincter (IAS; 0.5 cm) and E(m) was constant (-52 mV). Between the IAS and the proximal rectum an 18 mV gradient in E(m) developed across the muscle thickness with the submucosal edge at -70 mV and MPOs were replaced with slow waves (20 mV amplitude, 6 cpm). Slow waves were of greatest amplitude at the submucosal edge. Nifedipine (1 micro M) abolished MPOs but not slow waves. Contractile frequency changes were commensurate with the changes in pacemaker frequency. Our results suggest that changing motility patterns in the rectoanal region are associated with differences in the characteristics of pacemaker potentials as well as differences in the sites from which these potentials emanate. PMID- 12540369 TI - Double-stranded RNA activates a p38 MAPK-dependent cell survival program in biliary epithelia. AB - Double-stranded RNA (dsRNA) is produced during replicative viral infection or genotoxic stress. Thus knowledge of the cellular response to dsRNA is necessary to understand the effects of DNA damage or viral infection in biliary epithelia. We assessed the effect of dsRNA on biliary epithelial cell proliferation and apoptosis and the role of the stress-activated p38 MAPK signaling pathway in these responses. dsRNA did not induce apoptosis or proliferation in Mz-ChA-1 human malignant cholangiocytes, but decreased cytotoxicity induced by camptothecin or tumor necrosis factor-related apoptosis inducing ligand and decreased activity of caspases 3, 8, and 9. Furthermore, dsRNA increased p38 MAPK and JNK kinase active site phosphorylation but had no effect on either MAPK kinase (MEK)1/2 or protein kinase R phosphorylation. Inhibition of p38 MAPK with SB-203580 increased basal caspase activity. Thus dsRNA stimulates a p38 MAPK dependent cell-survival pathway in biliary epithelial cells that may modulate the response of the biliary epithelia to dsRNA produced during genotoxic injury or virus infection. PMID- 12540370 TI - Upregulation of activin A gene by butyrate in human colon cancer cell lines. AB - Activin A has been reported to play a role in the progression of colorectal cancer. Because dietary fiber protects against colorectal cancer, we hypothesized that butyrate, a fermentation product of dietary fiber, may affect the expression of activin A in colon cancer cells. Semiquantitative RT-PCR demonstrated that the activin A gene was upregulated by sodium butyrate in the human colon cancer cell lines HT-29 and Caco-2 in a concentration- and time-dependent manner. However, the activin A gene did not respond to sodium butyrate in the human normal colonic cell line FHC, rat normal intestinal epithelial cell (IEC) line IEC-6, and the explant of rat colon. Flow cytometry and agarose gel electrophoresis of genomic DNA revealed that cell cycle arrest and apoptosis were induced by sodium butyrate but not exogenous activin A in HT-29 cells, indicating that activin A could not act as an autocrine factor in colon cancer cells. By assuming that activin A promotes colorectal cancer spread as a paracrine factor, our findings suggest that butyrate could act as a tumor promoter in some circumstances. PMID- 12540371 TI - Regulation of copper absorption by copper availability in the Caco-2 cell intestinal model. AB - Relatively little is known about the individual steps in intestinal copper absorption and whether or how they may be regulated. Polarized Caco-2 cell monolayers with tight junctions offer an already tested model in which to study intestinal metal transport. This model was used to examine potential effects of cellular copper availability on copper absorption. Uptake and transport were determined on application of (64)Cu(II) to the brush border. In the range of 0.2 2 micro M, uptake was dose dependent and was approximately 20% of dose/90 min. Overall transport of (64)Cu across the basolateral surface was approximately 0.3%. When cellular copper levels were depleted 40% by 18-h pretreatment with the specific copper chelator triethylenetetraamine, uptake and overall transport were markedly increased, going to 80 and 65% of dose, respectively. Cellular retention of (64)Cu fell fourfold, from 6 to 1.5%. Depletion of copper with the chelator was rapid and preceded initial changes in uptake and overall transport by 4 h. A lesser depletion of cellular copper (13%) failed to enhance copper uptake but doubled the rate of overall transport, as measured with (64)Cu and by atomic absorption. As previously reported, preexposure of the cells to excess copper (10 micro M, 18 h) also enhanced copper uptake ( approximately 3-fold). In contrast, ascorbate (10-1,000 micro M) failed to significantly alter uptake and transport of 1 micro M (64)Cu. Our findings are consistent with the concepts that, in the low physiological range, copper availability alters the absorption capacity of the intestine to support whole body homeostasis and that basolateral transport is more sensitively regulated than uptake. PMID- 12540372 TI - Protein intake during hemodialysis maintains a positive whole body protein balance in chronic hemodialysis patients. AB - Protein energy malnutrition is present in 18 to 56% of hemodialysis patients. Because hemodialysis has been regarded as a catabolic event, we studied whether consumption of a protein- and energy-enriched meal improves the whole body protein balance during dialysis in chronic hemodialysis (CHD) patients. Patients were studied on a single day between dialysis (HD- protocol) in the morning while fasting and in the afternoon while consuming six small test meals. Patients were also studied during two separate dialysis sessions (HD+ protocol). Patients were fasted during one and consumed the meals during the other. Whole body protein metabolism was studied by primed constant infusion of l-[1-(13)C]valine. During HD-, feeding changed the negative whole body protein balance observed during fasting to a positive protein balance. Dialysis deepened the negative balance during fasting, whereas feeding during dialysis induced a positive balance comparable to the HD- protocol while feeding. Plasma valine concentrations during the studies were correlated with whole body protein synthesis and inversely correlated with whole body protein breakdown. We conclude that the consumption of a protein- and energy-enriched meal by CHD patients while dialyzing can strongly improve whole body protein balance, probably because of the increased amino acid concentrations in blood. PMID- 12540373 TI - Glucose-dependent insulinotropic polypeptide receptor null mice exhibit compensatory changes in the enteroinsular axis. AB - The incretins glucose-dependent insulinotropic polypeptide (GIP) and glucagon like peptide-1 (GLP-1) are gut hormones that act via the enteroinsular axis to potentiate insulin secretion from the pancreas in a glucose-dependent manner. Both GLP-1 receptor and GIP receptor knockout mice (GLP-1R(-/-) and GIPR(-/-), respectively) have been generated to investigate the physiological importance of this axis. Although reduced GIP action is a component of type 2 diabetes, GIPR deficient mice exhibit only moderately impaired glucose tolerance. The present study was directed at investigating possible compensatory mechanisms that take place within the enteroinsular axis in the absence of GIP action. Although serum total GLP-1 levels in GIPR knockout mice were unaltered, insulin responses to GLP 1 from pancreas perfusions and static islet incubations were significantly greater (40-60%) in GIPR(-/-) than in wild-type (GIPR(+/+)) mice. Furthermore, GLP-1-induced cAMP production was also elevated twofold in the islets of the knockout animals. Pancreatic insulin content and gene expression were reduced in GIPR(-/-) mice compared with GIPR(+/+) mice. Paradoxically, immunocytochemical studies showed a significant increase in beta-cell area in the GIPR-null mice but with less intense staining for insulin. In conclusion, GIPR(-/-) mice exhibit altered islet structure and topography and increased islet sensitivity to GLP-1 despite a decrease in pancreatic insulin content and gene expression. PMID- 12540374 TI - Effect of tumor removal on mucosal protein synthesis in patients with colorectal cancer. AB - It is currently controversial whether mucosal hyperproliferation is involved in colorectal cancerogenesis. The purpose of the present study was to examine protein synthetic rate as an indicator of potential tissue proliferation in grossly normal rectal mucosa from cancer-bearing subjects and to compare this rate with that in mucosa from subjects posttumor removal. Six postabsorptive patients with localized rectal cancer and five postsurgical control subjects received a primed constant infusion of [1-(13)C]leucine (0.16 micromol/kg min, 9.6 micromol/kg prime). Forceps biopsies from the mucosa were taken after 3 and 6 h. Protein synthesis was calculated from protein-bound leucine enrichment (determined by capillary GC-combustion IRMS) and from the enrichment of free intracellular leucine (determined by GC-quadrupole MS). In cancer-bearing subjects, mucosal protein synthesis amounted to 1.28 +/- 0.24%/h. This rate was significantly higher (P < 0.05) than the corresponding rate of mucosa from patients after cancer removal (0.69 +/- 0.09%/h). These findings do not support the concept that colorectal cancer originates from a proliferative disease of the whole colon. Increased mucosal protein synthesis appears to depend on the presence of the tumor itself and should therefore be considered a secondary phenomenon. PMID- 12540375 TI - Aberrant insulin-induced GLUT4 translocation predicts glucose intolerance in the offspring of a diabetic mother. AB - We examined the long-term effect of in utero exposure to streptozotocin-induced maternal diabetes on the progeny that postnatally received either ad libitum access to milk by being fed by control mothers (CM/DP) or were subjected to relative nutrient restriction by being fed by diabetic mothers (DM/DP) compared with the control progeny fed by control mothers (CM/CP). There was increased food intake, glucose intolerance, and obesity in the CM/DP group and diminished food intake, glucose tolerance, and postnatal growth restriction in the DM/DP group, persisting in the adult. These changes were associated with aberrations in hormonal and metabolic profiles and alterations in hypothalamic neuropeptide Y concentrations. By use of subfractionation and Western blot analysis techniques, the CM/DP group demonstrated a higher skeletal muscle sarcolemma-associated (days 1 and 60) and white adipose tissue plasma membrane-associated (day 60) GLUT4 in the basal state with a lack of insulin-induced translocation. The DM/DP group demonstrated a partial amelioration of this change observed in the CM/DP group. We conclude that the offspring of a diabetic mother with ad libitum postnatal nutrition demonstrates increased food intake and resistance to insulin-induced translocation of GLUT4 in skeletal muscle and white adipose tissue. This in turn leads to glucose intolerance and obesity at a later stage (day 180). Postnatal nutrient restriction results in reversal of this adult phenotype, thereby explaining the phenotypic heterogeneity that exists in this population. PMID- 12540376 TI - Epidermal growth factor upregulates beta-adrenergic receptor signaling in a human salivary cell line. AB - The effects of epidermal growth factor (EGF) on the beta-adrenergic receptor coupled adenylyl cyclase system were studied in a human salivary cell line (HSY). The beta-adrenergic agonist isoproterenol (10(-5) M) stimulated adenylyl cyclase activity by approximately 2-fold, and the isoproterenol response was increased 1.8-fold after prolonged (48 h) exposure to EGF (5 x 10(-10) M). In contrast, enzyme activation via stimulatory prostaglandin receptors and by agents acting on nonreceptor components of the adenylyl cyclase system was not enhanced by EGF. beta-Adrenergic receptor density, assessed by binding of the beta-adrenergic receptor antagonist (-)-[(125)I]iodopindolol, was increased threefold after EGF treatment. Competition binding studies with unlabeled antagonists selective for beta(1)- and beta(2)-adrenergic receptor subtypes indicated that the increase in (-)-[(125)I]iodopindolol binding sites induced by EGF reflected an increased number of beta(2)-adrenergic receptors. Likewise, Northern blot analysis of RNA from EGF-treated cells revealed selective induction of beta(2)-adrenergic receptor mRNA, which was blocked by the RNA synthesis inhibitor actinomycin D. The increase in beta-adrenergic receptor density produced by EGF was unaltered after phorbol ester-induced downregulation of protein kinase C (PKC). Enhancement of isoproterenol-responsive adenylyl cyclase activity and phosphorylation of mitogen-activated protein kinase (MAPK) by EGF were both blocked by the MAPK pathway inhibitor PD-98059. The results suggest that in HSY cells EGF enhances beta-adrenergic responsiveness by upregulating beta(2)-adrenergic receptor expression at the transcriptional level. Moreover, the stimulatory effect of EGF on beta(2)-adrenergic receptor signaling appears to be mediated by the MAPK pathway and independent of PKC activation. PMID- 12540377 TI - High glucose and endothelial cell growth: novel effects independent of autocrine TGF-beta 1 and hyperosmolarity. AB - Human endothelial cells were exposed to 5 mM glucose (control), 25 mM (high) glucose, or osmotic control for 72 h. TGF-beta1 production, cell growth, death, and cell cycle progression, and the effects of TGF-beta1 and TGF-beta neutralization on these parameters were studied. High glucose and hyperosmolarity increased endothelial TGF-beta1 secretion (P < 0.0001) and bioactivity (P < 0.0001). However, high glucose had a greater effect on reducing endothelial cell number (P < 0.001) and increasing cellular protein content (P < 0.001) than the osmotic control. TGF-beta antibody only reversed the antiproliferative and hypertrophic effects of high glucose. High glucose altered cell cycle progression and cyclin-dependent kinase inhibitor expression independently of hyperosmolarity. High glucose increased endothelial cell apoptosis (P < 0.01), whereas hyperosmolarity induced endothelial cell necrosis (P < 0.001). TGF-beta antibody did not reverse the apoptotic effects observed with high glucose. Exogenous TGF-beta1 mimicked the increased S phase delay but not endoreduplication observed with high glucose. High glucose altered endothelial cell growth, apoptosis, and cell cycle progression. These growth effects occurred principally via a TGF-beta1 autocrine pathway. In contrast, apoptosis and endoreduplication occurred independently of this cytokine and hyperosmolarity. PMID- 12540378 TI - A negatively charged residue in the outer mouth of rat sodium channel determines the gating kinetics of the channel. AB - Previous studies using combined techniques of site-directed mutagenesis and electrophysiology of voltage-gated Na(+) channels have demonstrated that there are significant overlaps in the regions that are important for the two fundamental properties of the channels, namely gating and permeation. We have previously shown that a pore-lining residue, W402 in S5-S6 region (P loop) in domain I of the micro1 skeletal muscle Na(+) channel, was important in the gating of the channel. Here, we determined the role of an adjacent pore-lining negatively charged residue (E403) in channel gating. Charge neutralization or substitution with positively charged side chain at this position resulted in a marked delay in the rate of recovery from slow inactivation. Indeed, the fast inactivation process appeared intact. Restoration of the negatively charged side chain with a sulfhydryl modifier, MTS-ethylsulfonate, resulted in a reactivation profile from a slow-inactivated state, which was indistinguishable from that of the wild-type channels. We propose an additional functional role for the negatively charged residue. Assuming no major changes in the pore structure induced by the mutations, the negatively charged residue E403 may work in concert with other pore regions during recovery from slow inactivation of the channel. Our data represent the first report indicating the role of negative charge in the slow inactivation of the voltage-gated Na(+) channel. PMID- 12540379 TI - Cell volume increase and extracellular Ca2+ are needed for hyposmotically induced prolactin release in tilapia. AB - In the tilapia (Oreochromis mossambicus), as in many euryhaline teleost fish, prolactin (PRL) plays a central role in freshwater adaptation, acting on osmoregulatory surfaces to reduce ion and water permeability and increase solute retention. Consistent with these actions, PRL release is stimulated as extracellular osmolality is reduced both in vivo and in vitro. In the current experiments, a perfusion system utilizing dispersed PRL cells was developed for permitting the simultaneous measurement of cell volume and PRL release. Intracellular Ca(2+) was monitored using fura 2-loaded cells under the same conditions. When PRL cells were exposed to hyposmotic medium, an increase in PRL cell volume preceded the increase in PRL release. Cell volume increased in proportion to decreases of 15 and 30% in osmolality. However, regulatory volume decrease was clearly seen only after a 30% reduction. The hyposmotically induced PRL release was sharply reduced in Ca(2+)-deleted hyposmotic medium, although cell volume changes were identical to those observed in normal hyposmotic medium. In most cells, a rise in intracellular Ca(2+) concentration ([Ca(2+)](i)) during hyposmotic stimulation was dependent on the availability of extracellular Ca(2+), although small transient increases in [Ca(2+)](i) were sometimes observed upon introduction of Ca(2+)-deleted media of the same or reduced osmolality. These results indicate that an increase in cell size is a critical step in the transduction of an osmotic signal into PRL release and that the hyposmotically induced increase in PRL release is greatly dependent on extracellular Ca(2+). PMID- 12540380 TI - Evidence that signal transduction for osmoreception is mediated by stretch activated ion channels in tilapia. AB - Prolactin (PRL) plays a central role in the freshwater osmoregulation of teleost fish, including the tilapia (Oreochromis mossambicus). Consistent with this action, PRL release from the tilapia pituitary increases as extracellular osmolality is reduced both in vitro and in vivo. Dispersed tilapia PRL cells were incubated in a perfusion chamber that allowed simultaneous measurements of cell volume and PRL release. Intracellular Ca(2+) concentrations were measured from fura 2-loaded PRL cells treated in a similar way. Gadolinium (Gd(3+)), known to block stretch-activated cation channels, inhibited hyposmotically induced PRL release in a dose-related manner without preventing cell swelling. Nifedipine, an L-type Ca(2+) channel blocker, did not prevent the increase in PRL release during hyposmotic stimulation. A high, depolarizing concentration of KCl induced a transient and marked increase of intracellular Ca(2+) and release of PRL but did not prevent the rise in intracellular Ca(2+) and PRL release evoked by exposure to hyposmotic medium. These findings suggest that a decrease in extracellular osmolality stimulates PRL release through the opening of stretch-activated ion channels, which allow extracellular Ca(2+) to enter the cell when it swells. PMID- 12540381 TI - Rab5a and rab11a mediate agonist-induced trafficking of protease-activated receptor 2. AB - We evaluated the contribution of rab5a and rab11a to trafficking and signaling of protease-activated receptor 2 (PAR2), a receptor for trypsin and tryptase. Agonists stimulated internalization of PAR2 into early endosomes containing rab5a. Dominant negative rab5aS34N disrupted early endosomes and inhibited agonist-stimulated endocytosis of PAR2. Internalized PAR2 was sorted to lysosomes, and rab5a remained in early endosomes. Rab5a promoted and rab5aS34N impeded resensitization of trypsin-induced calcium mobilization. Rab11a was detected in the Golgi apparatus with PAR2, and PAR2 agonists stimulated redistribution of rab11a into vesicles containing PAR2 that migrated to the cell surface. Dominant negative rab11aS25N was mostly confined to the Golgi apparatus. Although expression of rab11aS25N caused retention of PAR2 in the Golgi apparatus, it did not abolish trafficking of PAR2 to the cell surface. However, expression of wild-type rab11a accelerated both recovery of PAR2 at the cell surface and resensitization of PAR2 signaling. Thus rab5a is required for PAR2 endocytosis and resensitization, whereas rab11a contributes to trafficking of PAR2 from the Golgi apparatus to the plasma membrane. PMID- 12540382 TI - n-3 Long-chain polyunsaturated fatty acids reduce risk of coronary heart disease death: extending the evidence to the elderly. PMID- 12540383 TI - Calcium intake and adiposity. AB - Limited epidemiologic and experimental data support the possibility that dietary calcium intake plays a role in human body weight regulation. The aim of this review was to present the data from human studies that link calcium and dairy intake to body weight, describe the existing evidence for an effect of calcium intake on body weight from animal models of obesity, present evidence of a role for intracellular calcium in the regulation of lipogenesis and lipolysis, elucidate the potential suggested relation between dietary calcium intake and intracellular calcium concentrations, and outline the effects of calcium supplementation on dietary fat absorption. We suggest that these data support the need for large, population based clinical trials to assess the effects of supplemental calcium and other components of dairy products on human body weight. PMID- 12540384 TI - Denon Institute Award for Excellence in Medical/Dental Nutrition Education Lecture, 2002. Will there be a tipping point in medical nutrition education? AB - The title of this presentation is derived from a national bestseller that defines the moment when an idea, trend, or social behavior crosses a threshold, "tips," and becomes widely disseminated. Three key tipping-point factors are discussed in the context of medical nutrition education. The Law of Few has to do with the essentiality of role models in directing educational curricula and clinical programs. The Stickiness Factor describes the quality and the content of the message that enable the information to have prolonged meaning. The Power of Context refers to the conditions and risk factors that are directly affected by diet and nutrition. Examples of these key factors are provided with a look to the future of medical nutrition education. PMID- 12540385 TI - Insulinogenic index at 15 min as a marker of nutritional rehabilitation in anorexia nervosa. AB - BACKGROUND: Insulin responses to the oral-glucose-tolerance test (OGTT) in anorexia nervosa (AN) are related to body weight and show various patterns. Although weight gain is a key indicator of a successful nutritional program, it is not a sufficiently accurate index for assessing nutritional status, especially in the periods of marked fear of obesity, because patients often manipulate body weight measurements. OBJECTIVE: The aim of this study was to determine the relation between insulin metabolism during the early phase of the OGTT and progress (weekly weight gain) during nutritional rehabilitation. DESIGN: Forty eight inpatients with AN (25 AN restricting type and 23 AN bulimic type) underwent the OGTT, with additional blood sampling at 15 min, when energy intake reached 6694 kJ/d (1600 kcal/d). Thirteen healthy volunteers were also studied. To evaluate early-phase insulin metabolism, we calculated the insulinogenic index after 15 (II(15 min)) and 30 min. On the basis of weekly changes in body weight, the AN participants were divided into good (> or =0.5 kg) and poor (<0.5 kg) responders. RESULTS: Among the AN patients, 48% were poor responders. Analysis of variance showed significant differences in the II(15 min) values (P = 0.0005) and showed that II(15 min) values for good responders were significantly higher than those for the other groups. CONCLUSIONS: These findings suggest that a lack of progress in weight gain is frequently observed in AN and that II(15 min) values may be a useful marker with which to assess the weekly progress during nutritional rehabilitation. PMID- 12540386 TI - Randomized controlled trial of the effect of n-3 fatty acid supplementation on the metabolism of apolipoprotein B-100 and chylomicron remnants in men with visceral obesity. AB - BACKGROUND: Lipid abnormalities may contribute to the increased risk of atherosclerosis and coronary disease in visceral obesity. Fish oils lower plasma triacylglycerols, but the underlying mechanisms are not fully understood. OBJECTIVE: We studied the effect of fish oils on the metabolism of apolipoprotein B-100 (apo B) and chylomicron remnants in obese men. DESIGN: Twenty-four dyslipidemic, viscerally obese men were randomly assigned to receive either fish oil capsules (4 g/d, consisting of 45% eicosapentaenoic acid and 39% docosahexaenoic acid as ethyl esters) or matching placebo (corn oil, 4 g/d) for 6 wk. VLDL, intermediate-density lipoprotein (IDL), and LDL apo B kinetics were assessed by following apo B isotopic enrichment with the use of gas chromatography-mass spectrometry after an intravenous bolus injection of trideuterated leucine. Chylomicron remnant catabolism was measured with the use of an intravenous injection of a chylomicron remnant-like emulsion containing cholesteryl [(13)C]oleate, and isotopic enrichment of (13)CO(2) in breath was measured with isotope ratio mass spectrometry. Kinetic values were derived with multicompartmental models. RESULTS: Fish oil supplementation significantly (P < 0.05) lowered plasma concentrations of triacylglycerols (-18%) and VLDL apo B ( 20%) and the hepatic secretion of VLDL apo B (-29%) compared with placebo. The percentage of conversions of VLDL apo B to IDL apo B, VLDL apo B to LDL apo B, and IDL apo B to LDL apo B also increased significantly (P < 0.05): 71%, 93%, and 11%, respectively. Fish oils did not significantly alter the fractional catabolic rates of apo B in VLDL, IDL, or LDL or alter the catabolism of the chylomicron remnant-like emulsion. CONCLUSION: Fish oils effectively lower the plasma concentration of triacylglycerols, chiefly by decreasing VLDL apo B production but not by altering the catabolism of apo B-containing lipoprotein or chylomicron remnants. PMID- 12540387 TI - Reduced hemodynamic load and cardiac hypotrophy in patients with anorexia nervosa. AB - BACKGROUND: Anorexia nervosa is associated with lower left ventricular mass (LVM) and systolic dysfunction. Whether these abnormalities reflect chronic protein energy malnutrition or are primarily related to lower cardiac workload is unclear. OBJECTIVE: The objective of the study was to verify whether low LVM in anorexia nervosa is explained by low hemodynamic load. DESIGN: Ninety-one women with anorexia nervosa [macro x +/- SD age: 20.5 +/- 6.1 y; body mass index (in kg/m(2)): 15.6 +/- 1.9; group 1] and 62 normal-weight female control subjects (age: 22.5 +/- 5.5 y; body mass index: 20.9 +/- 1.2; group 2) underwent Doppler echocardiography. LVM was evaluated as the percentage predicted by body height, sex, and stroke work (systolic blood pressure x stroke volume). RESULTS: The left ventricular chamber dimension was smaller and the chamber walls were thinner in group 1 than in group 2, which resulted in significantly lower LVM and LVM indexes (P < 0.0001). Ejection fraction, heart rate, stroke volume, and cardiac output were significantly (P < 0.007) lower in group 1, but peripheral resistance was substantially higher (P < 0.0001). The deviation of LVM from predicted values was lower and the proportion of subjects with inadequate LVM was significantly higher in group 1 than in group 2 (P < 0.0001). This difference was attenuated after adjustment for body weight and heart rate. There were no relations between LVM and laboratory tests in group 1. CONCLUSIONS: Anorexia nervosa is a condition of low hemodynamic load that leads to low LVM. Even with adjustment for stroke work, however, LVM is lower than would be predicted by height, because of the effect of body weight reduction (ie, wasting of lean body mass). PMID- 12540388 TI - A short-term, high-fat diet up-regulates lipid metabolism and gene expression in human skeletal muscle. AB - BACKGROUND: Dietary fatty acids may be important in regulating gene expression. However, little is known about the effect of changes in dietary fatty acids on gene regulation in human skeletal muscle. OBJECTIVE: The objective was to determine the effect of altered dietary fat intake on the expression of genes encoding proteins necessary for fatty acid transport and beta-oxidation in skeletal muscle. DESIGN: Fourteen well-trained male cyclists and triathletes with a mean (+/- SE) age of 26.9 +/- 1.7 y, weight of 73.7 +/- 1.7 kg, and peak oxygen uptake of 67.0 +/- 1.3 mL x kg(-1) x min(-1) consumed either a high-fat diet (HFat: > 65% of energy as lipids) or an isoenergetic high-carbohydrate diet (HCho: 70-75% of energy as carbohydrate) for 5 d in a crossover design. On day 1 (baseline) and again after 5 d of dietary intervention, resting muscle and blood samples were taken. Muscle samples were analyzed for gene expression [fatty acid translocase (FAT/CD36), plasma membrane fatty acid binding protein (FABPpm), carnitine palmitoyltransferase I (CPT I), beta-hydroxyacyl-CoA dehydrogenase (beta-HAD), and uncoupling protein 3 (UCP3)] and concentrations of the proteins FAT/CD36 and FABPpm. RESULTS: The gene expression of FAT/CD36 and beta -HAD and the gene abundance of FAT/CD36 were greater after the HFat than after the HCho diet (P < 0.05). Messenger RNA expression of FABPpm, CPT I, and UCP-3 did not change significantly with either diet. CONCLUSIONS: A rapid and marked capacity for changes in dietary fatty acid availability to modulate the expression of mRNA encoding proteins is necessary for fatty acid transport and oxidative metabolism. This finding is evidence of nutrient-gene interactions in human skeletal muscle. PMID- 12540389 TI - n-3 Polyunsaturated fatty acids, fatal ischemic heart disease, and nonfatal myocardial infarction in older adults: the Cardiovascular Health Study. AB - BACKGROUND: Little is known about the relation of the dietary intake of n-3 polyunsaturated fatty acids, ie, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) from fatty fish and alpha-linolenic acid from vegetable oils, with ischemic heart disease among older adults. OBJECTIVE: We investigated the associations of plasma phospholipid concentrations of DHA, EPA, and alpha linolenic acid as biomarkers of intake with the risk of incident fatal ischemic heart disease and incident nonfatal myocardial infarction in older adults. DESIGN: We conducted a case-control study nested in the Cardiovascular Health Study, a cohort study of adults aged > or = 65 y. Cases experienced incident fatal myocardial infarction and other ischemic heart disease death (n = 54) and incident nonfatal myocardial infarction (n = 125). Matched controls were randomly selected (n = 179). We measured plasma phospholipid concentrations of n-3 polyunsaturated fatty acids in blood samples drawn approximately 2 y before the event. RESULTS: A higher concentration of combined DHA and EPA was associated with a lower risk of fatal ischemic heart disease, and a higher concentration of alpha-linolenic acid with a tendency to lower risk, after adjustment for risk factors [odds ratio: 0.32 (95% CI: 0.13, 0.78; P = 0.01) and 0.52 (0.24, 1.15; P = 0.1), respectively]. In contrast, n-3 polyunsaturated fatty acids were not associated with nonfatal myocardial infarction. CONCLUSIONS: Higher combined dietary intake of DHA and EPA, and possibly alpha-linolenic acid, may lower the risk of fatal ischemic heart disease in older adults. The association of n-3 polyunsaturated fatty acids with fatal ischemic heart disease, but not with nonfatal myocardial infarction, is consistent with possible antiarrhythmic effects of these fatty acids. PMID- 12540390 TI - A fermented milk high in bioactive peptides has a blood pressure-lowering effect in hypertensive subjects. AB - BACKGROUND: Angiotensin-converting enzyme (ACE; EC 3.4.15.1) plays a dual role in the regulation of hypertension: it catalyzes the production of the vasoconstrictor angiotensin II and it inactivates the vasodilator bradykinin. By inhibiting these processes, ACE inhibitors have antihypertensive effects. Peptides derived from milk proteins can have ACE-inhibiting properties and may thus be used as antihypertensive components. OBJECTIVE: We evaluated the long term blood pressure-lowering effect of milk fermented by Lactobacillus helveticus LBK-16H in hypertensive subjects. DESIGN: In a randomized placebo-controlled study, 39 hypertensive patients received 150 mL/d of either L. helveticus LBK-16H fermented milk or a control product for 21 wk after a 2-wk run-in period. During the run-in period, the average baseline diastolic and systolic blood pressure values were 155 and 97 mm Hg, respectively, in the test product group and 152 and 96 mm Hg, respectively, in the control group. After the run-in period, blood pressure was measured at home on the same day every week with the use of an automatic blood pressure recorder. RESULTS: There was a mean difference of 6.7 +/ 3.0 mm Hg in systolic blood pressure (P = 0.030) and of 3.6 +/- 1.9 mm Hg (P = 0.059) in diastolic blood pressure between the test product and control groups. Demographic factors had no significant effect on the responses. CONCLUSION: L. helveticus LBK-16H fermented milk containing bioactive peptides in normal daily use has a blood pressure-lowering effect in hypertensive subjects. PMID- 12540391 TI - Development of bioelectrical impedance analysis prediction equations for body composition with the use of a multicomponent model for use in epidemiologic surveys. AB - BACKGROUND: Previous studies to develop and validate bioelectrical impedance analysis (BIA) equations to predict body composition were limited by small sample sizes, sex specificity, and reliance on reference methods that use a 2-component model. OBJECTIVE: This study was designed to develop sex-specific BIA equations to predict total body water (TBW) and fat-free mass (FFM) with the use of a multicomponent model for children and adults. DESIGN: Data from 5 centers were pooled to create a sample of 1474 whites and 355 blacks aged 12-94 y. TBW was measured by dilution, and FFM was estimated with a multicomponent model based on densitometry, isotope dilution, and dual-energy X-ray absorptiometry. RESULTS: The final race-combined TBW prediction equations included stature(2)/resistance and body weight (R(2) = 0.84 and 0.79 and root mean square errors of 3.8 and 2.6 L for males and females, respectively; CV: 8%) and tended to underpredict TBW in black males (2.0 L) and females (1.4 L) and to overpredict TBW in white males (0.5 L) and females (0.3 L). The race-combined FFM prediction equations contained the same independent variables (R(2) = 0.90 and 0.83 and root mean square errors of 3.9 and 2.9 kg for males and females, respectively; CV: approximately 6%) and tended to underpredict FFM in black males (2.1 kg) and females (1.6 kg) and to overpredict FFM in white males (0.4 kg) and females (0.3 kg). CONCLUSION: These equations have excellent precision and are recommended for use in epidemiologic studies to describe normal levels of body composition. PMID- 12540392 TI - Body mass index, height, weight, arm circumference, and mortality in rural Bangladeshi women: a 19-y longitudinal study. AB - BACKGROUND: Studies in Western populations report a J- or U-shaped relation between body mass index (BMI; in kg/m(2)) and mortality, in which persons with extremes of BMI experience increased mortality. In contrast, little is known about populations in developing countries, where nutritional status is lower. OBJECTIVE: The objective was to examine the association between BMI and mortality in Bangladeshi women. DESIGN: A cohort of 1888 rural Bangladeshi women (mean age: 27.9 y) was followed over 19 y. Height, weight, arm circumference, fertility, and socioeconomic data were obtained between 1975 and 1979. Mortality, loss-to-follow up, and additional socioeconomic data were identified by the demographic surveillance system of the International Centre for Health and Population Research, Bangladesh. Proportional hazards regression was used to examine the relation between BMI and all-cause mortality. RESULTS: The association between BMI and mortality was reverse J-shaped. After adjustment for socioeconomic indicators, the risk of dying was highest in women with BMIs in the lowest 10% of the decile distribution (< 16.39) and lowest in women with intermediate (11-89% range of the decile distribution) BMIs (16.39-20.71). Women with BMIs in the highest 10% of the distribution (> 20.71) had slightly elevated mortality (NS) compared with those with intermediate BMIs. Age and education were strongly associated with mortality. Women without schooling had a risk of mortality 4 times that of women with > or = 1 y of schooling. CONCLUSIONS: A woman's BMI relative to the BMI distribution in the local population may be a better predictor of mortality than is absolute BMI. The contribution of education in reducing mortality supports development programs aimed at increasing women's education. PMID- 12540393 TI - Human skin condition and its associations with nutrient concentrations in serum and diet. AB - BACKGROUND: Nutritional factors exert promising actions on the skin, but only scant information is available on the modulating effects of physiologic concentrations of nutrients on the skin condition of humans. OBJECTIVE: The objective was to evaluate whether nutrient concentrations in serum and diet are associated with the skin condition of humans. DESIGN: A cross-sectional study was conducted in which data on serum concentrations of nutrients, dietary intake of nutrients, and the hydration, sebum content, and surface pH of skin were obtained from 302 healthy men and women. Skin condition was measured with the use of noninvasive techniques. Dietary intake was assessed with 2 complementary food frequency questionnaires. Multiple regression analysis was used to evaluate associations of serum vitamins and carotenoids and of dietary micro- and macronutrients with skin condition. RESULTS: After adjustment for potential confounders, including sex, age, and smoking, statistically significant associations were shown in the total population between serum vitamin A and skin sebum content and surface pH and between the dietary intake of total fat, saturated fat, monounsaturated fat, and skin hydration. Monounsaturated fat intake was also associated with surface pH. Associations between serum beta cryptoxanthin and skin hydration and between surface pH and fluid and calcium intakes were observed in men only. CONCLUSION: Several associations between nutrients in serum and diet and skin condition were observed, indicating that changes in baseline nutritional status may affect skin condition. PMID- 12540394 TI - Comparison of the effectiveness of 2 dual-energy X-ray absorptiometers with that of total body water and computed tomography in assessing changes in body composition during weight change. AB - BACKGROUND: Little information is available on the assessment of changes in body composition as a function of weight change with the use of the fan beam of dual energy X-ray absorptiometry (DXA). OBJECTIVE: The objective was to determine the accuracy of the fan beam of the QDR 4500A densitometer and the pencil beam of the QDR 2000 densitometer in estimating changes in whole-body lean soft tissue mass (LSTM(DXA)) and fat mass (FM) with weight change. DESIGN: Thirty-seven subjects who lost 5.7 +/- 4.5 kg were measured before and after weight change. Using total body water and computed tomography (CT) of the midthigh, we compared changes in FFM(TBW) and LSTM(CT) with changes in LSTM(DXA). RESULTS: Overall, compared with TBW, the fan beam gave a larger estimate of change (macro x +/- SD) in LSTM (fan beam - TBW: -0.7 +/- 1.6 kg) than did the pencil beam (pencil beam - TBW: -0.1 +/ 1.6 kg). When the change in LSTM obtained with the fan beam and pencil beam was regressed against the change in FFM(TBW), the slope of the line for the fan beam was 0.97 (r(2) = 0.61) and that for the pencil beam was 0.86 (r(2) = 0.61). Regression analysis showed that the results between the 2 units were not interchangeable. For the midthigh region, the change in LSTM(CT) was moderately correlated with the change in LSTM(DXA) with the fan beam and pencil beam. CONCLUSIONS: The measurement of change in LSTM with the fan and pencil beams provides the same relation to changes in FFM assessed by TBW, but the 2 systems are not interchangeable. PMID- 12540395 TI - Evaluation of a new pediatric air-displacement plethysmograph for body composition assessment by means of chemical analysis of bovine tissue phantoms. AB - BACKGROUND: Body-composition assessment reflects infant growth and nutritional status but is limited by practical considerations, accuracy, and safety. OBJECTIVE: This study evaluated the precision and accuracy of a new air displacement plethysmography (ADP) system for pediatric body-composition assessment. DESIGN: We used 24 phantoms constructed from bovine lean muscle and fat. The phantoms varied in mass (1.3894-9.9516 kg) and percentage fat (%Fat; 2.08-34.40%), thereby representing infants between birth and 6 mo of age. Estimates of %Fat obtained with chemical analysis (CA), hydrostatic weighing, and ADP were compared. RESULTS: There was no significant difference between %Fat measured with ADP (%Fat(ADP)) and %Fat measured with CA (%Fat(CA)); the mean values were 18.55% and 18.59%, respectively. SDs for %Fat(ADP) and %Fat(CA) were not significantly different (0.70% and 0.73%, respectively). %Fat measurements obtained with ADP, CA, and hydrostatic weighing were highly correlated (r > 0.99, P < 0.0001). The regression equation (%Fat(CA) = 0.996%Fat(ADP) + 0.119; SEE = 0.600; adjusted R(2) = 0.997; P < 0.0001) did not differ significantly from the line of identity (%Fat(CA) = %Fat(ADP)). There was high agreement between individual measurements of %Fat(ADP) and %Fat(CA), as shown by the narrow 95% limits of agreements between methods (-1.22% to 1.13%), and there was no systematic bias in individual differences across the phantom mass and %Fat ranges. CONCLUSION: ADP provides a highly precise and accurate estimate of %Fat in bovine tissue phantoms in the pediatric ranges of body weight and body fatness. PMID- 12540396 TI - Differences in resting metabolic rate between paraplegic and able-bodied subjects are explained by differences in body composition. AB - BACKGROUND: Little is known about the relation between body composition and energy metabolism in paraplegia. OBJECTIVE: We investigated the relation between body composition and energy metabolism in healthy paraplegics as compared with able-bodied control subjects. We hypothesized that paraplegics would have lower fat-free mass (FFM), body cell mass (BCM), resting metabolic rate (RMR), and thermic effect of feeding (TEF). DESIGN: This cross-sectional study included 34 control subjects and 28 paraplegics (mean age: 29.1 +/- 7.6 and 33.9 +/- 9.2 y, respectively) with body mass indexes (in kg/m(2)) of 23.5 +/- 1.8 and 24.3 +/- 6.0, respectively. We measured RMR and TEF with indirect calorimetry, total body water with deuterium dilution, and extracellular water with corrected bromide space. We calculated FFM (total body water/0.732) and BCM [(total body water - extracellular water)/0.732)]. RESULTS: FFM was higher in control subjects than in paraplegics (77.2 +/- 7.2% and 69.2 +/- 8.7%, respectively; P = 0.0002), as were BCM (47.4 +/- 6.7% and 35.9 +/- 8.1%, respectively; P < 0.0001) and RMR (7016 +/- 935 and 6159 +/- 954 kJ/d, respectively; P = 0.0007). FFM was the single best predictor of RMR in both groups (r(2) = 0.83 for control subjects and 0.70 for paraplegics, P < 0.0001 for both). RMR adjusted for FFM did not differ significantly between control subjects and paraplegics (6670 +/- 504 and 6588 +/- 501 kJ/d, respectively). TEF also did not differ significantly between control subjects and paraplegics (6.25 +/- 2.2% and 5.53 +/- 1.8% of energy intake, respectively). CONCLUSIONS: FFM, BCM, and RMR, but not obligatory TEF, are lower in paraplegics than in control subjects. RMR does not differ between control and paraplegic subjects after adjustment for FFM, indicating similar metabolic activity in the fat-free compartment of the body. PMID- 12540397 TI - Comparisons of waist circumferences measured at 4 sites. AB - BACKGROUND: Waist circumference (WC) is now accepted as a practical measure of adipose tissue distribution. Four body sites for WC measurements are commonly used, as follows: immediately below the lowest ribs (WC1), the narrowest waist (WC2), the midpoint between the lowest rib and the iliac crest (WC3), and immediately above the iliac crest (WC4). OBJECTIVE: We sought to compare the magnitude and reliability of WC measured at these 4 sites in males and females. DESIGN: WC was measured at each site 1 time in all subjects [49 males and 62 females, aged 7-83 y, with a body mass index (in kg/m(2)) of 9-43] and 3 times in a subgroup (n = 93) by one experienced observer using a heavy-duty inelastic tape. Body fat was measured in a subgroup (n = 74) with the use of dual-energy X ray absorptiometry. RESULTS: The mean values of WC were WC2 < WC1 < WC3 < WC4 (P < 0.01) in females and WC2 < WC1, WC3, and WC4 (P < 0.01) in males. For all 4 sites, measurement reproducibility was high, with intraclass correlation (r) values > 0.99. WC values were significantly correlated with fatness; correlations with trunk fat were higher than correlations with total body fat in both sexes. CONCLUSIONS: WC values at the 4 commonly used anatomic sites differ in magnitude depending on sex, are highly reproducible, and are correlated with total body and trunk adiposity in a sex-dependent manner. These observations have implications for the use of WC measurements in clinical practice and patient-oriented research. PMID- 12540398 TI - High-fiber rye bread and insulin secretion and sensitivity in healthy postmenopausal women. AB - BACKGROUND: Fiber and whole-cereal intakes may protect against hyperinsulinemia and the risk of type 2 diabetes. OBJECTIVE: The aim was to study whether the long term use of high-fiber rye bread and white-wheat bread modifies glucose and insulin metabolism in healthy postmenopausal women. DESIGN: The study was a randomized crossover trial consisting of 8-wk test and 8-wk washout periods. The subjects were 20 postmenopausal women [macro x +/- SD age: 59 +/- 6.0 y; body mass index (in kg/m(2)): 27.5 +/- 2.9; baseline fasting serum cholesterol: 6.5 +/ 0.8 mmol/L], of whom 3 had impaired glucose tolerance as determined by a 2-h oral-glucose-tolerance test. The test breads were high-fiber rye and white-wheat breads, planned to make up > or =20% of energy. Fasting blood samples were collected for the measurement of plasma glucose and insulin at the beginning and at the end of both bread periods. The frequently sampled intravenous-glucose tolerance test was performed at the run-in and at the end of both bread periods. The acute insulin response, insulin sensitivity, and glucose effectiveness were calculated. RESULTS: The rye bread made up 23.4 +/- 4.3% and wheat bread 26.7 +/- 8.2% of total energy intake. Compared with that during the run-in period, the acute insulin response increased significantly more during the rye bread period (9.9 +/- 24.2%) than during the wheat bread period (2.8 +/- 36.3%; P = 0.047). Other measured variables did not change significantly during the study. CONCLUSIONS: Modification of carbohydrate intake by high-fiber rye bread did not alter insulin sensitivity in postmenopausal, hypercholesterolemic women. High fiber rye bread appears to enhance insulin secretion, possibly indicating improvement of b cell function. PMID- 12540399 TI - Leptin and energy metabolism in pulmonary tuberculosis. AB - BACKGROUND: Pulmonary tuberculosis is the classic cause of "consumption," but the pathogenesis of such wasting is largely unknown. Animal studies in other conditions suggest that leptin may be a mediator between proinflammatory cytokine activity and wasting. OBJECTIVE: We tested whether the leptin concentration, after control for body fat mass, is higher during active pulmonary tuberculosis than after recovery and whether it correlates with energy metabolism and proinflammatory cytokine activity. DESIGN: Nondiabetic adults with pulmonary tuberculosis (n = 32) were recruited into a prospective observational study. Patients found to be antibody positive for human immunodeficiency virus were excluded from the study. Dual-energy X-ray absorptiometry, indirect calorimetry, and food intake protocols were performed at baseline and after 1 and 6 mo of tuberculosis treatment. Fasting plasma leptin, tumor necrosis factor alpha and its soluble receptor, and interleukin 6 were measured by enzyme-linked immunosorbent assay. RESULTS: Resting energy expenditure was close to Harris Benedict predictions and did not change significantly during treatment, but energy intake increased. Leptin concentration was correlated in a log-linear fashion with percentage body fat but was independent of cytokines and energy intake. There was no significant difference in leptin, corrected for energy balance and fat mass, at baseline and after 1 and 6 mo of treatment. CONCLUSIONS: These data are compatible with recovery from anorexia or starvation without discernible hyper- or hypometabolism. The close correlation of leptin with body fat mass is similar to observations in healthy subjects. No additional influence of disease state or proinflammatory cytokine activity was found. Leptin does not appear to be a component of the immune response to human pulmonary tuberculosis, and thus it cannot account for the weight loss and anorexia associated with tuberculosis. PMID- 12540400 TI - A novel dual radio- and stable-isotope method for measuring calcium absorption in humans: comparison with the whole-body radioisotope retention method. AB - BACKGROUND: Dietary calcium absorption can be determined only with the use of isotope techniques. Currently used isotope techniques require exclusive equipment or are not true tracer approaches. OBJECTIVE: The objective was to compare a dual isotope method combining radioisotopes and stable isotopes with a whole-body radioisotope retention method for measuring calcium absorption. DESIGN: Seven healthy adults aged 21-27 y consumed a test meal containing 63 +/- 14 (macro x +/ SD) mg Ca together with a water solution of (47)Ca (0.11 MBq). One hour after ingestion, 18 mg (44)Ca was administered intravenously. All feces and urine were collected for 5 and 6 d, respectively. Calcium absorption was estimated from whole-body retention of the radioisotope 12 times over 3 wk after ingestion and from the excretion of (47)Ca and (44)Ca in a 24-h urine sample collected on day 2. (44)Ca in urine was determined by inductively coupled plasma mass spectrometry. RESULTS: Mean (+/- SD) calcium absorption was 75 +/- 9% with the dual-isotope method and was 74 +/- 8% with the whole-body radioisotope retention method. There was a high degree of agreement between the methods. CONCLUSION: The dual-isotope method is a valid approach for measuring calcium absorption from a single meal. PMID- 12540401 TI - Diurnal variations in human urinary excretion of nicotinamide catabolites: effects of stress on the metabolism of nicotinamide. AB - BACKGROUND: More than 500 enzymes need niacin coenzymes. Therefore, elucidation of the control mechanisms of coenzyme metabolism is fundamentally important. OBJECTIVE: NAD(+) is involved in ATP production. Because energy expenditure is generally higher during the day than at night, we investigated whether the metabolism of nicotinamide changes at various times of day and whether stress affects nicotinamide metabolism. DESIGN: Twelve women were housed in the same facility and followed the same schedule for activities of daily living for 12 d. Urinary outputs were collected during 5 specific periods to investigate diurnal variations in nicotinamide metabolism. The effects of cold exposure (physical stress), having to perform arithmetic calculations (mental stress), and dark exposure (emotional stress) on nicotinamide metabolism were investigated. RESULTS: A diurnal variation in the nicotinamide metabolites N(1) methylnicotinamide, N(1)-methyl-2-pyridone-5-carboxamide, and N(1)-methyl-4 pyridone-3-carboxamide was observed. Of the stresses studied, cold exposure significantly increased the urinary excretory outputs of the nicotinamide metabolites. CONCLUSIONS: Diurnal variations in nicotinamide metabolism were found in these women. The biosynthesis of nicotinamide from tryptophan seemed to be increased by cold exposure. PMID- 12540402 TI - Comparing the pharmacokinetics of daidzein and genistein with the use of 13C labeled tracers in premenopausal women. AB - BACKGROUND: Despite significant interest in the risks and benefits of phytoestrogens to human health, few data exist on their pharmacokinetics in humans. OBJECTIVE: We investigated the pharmacokinetics of the (13)C isotopic forms of daidzein and genistein in healthy humans, specifically addressing intraindividual variability, effect of increasing intake, and influence of prolonged exposure to a soy food diet. DESIGN: Premenopausal women (n = 16) were administered 0.4 mg [(13)C]daidzein or [(13)C]genistein/kg body wt orally on 3 occasions, including once after eating soy foods for 7 d. On a further occasion the dose was doubled. Plasma and urinary [(13)C]isoflavone concentrations were measured by mass spectrometry. RESULTS: Serum concentrations of [(13)C]genistein and [(13)C]daidzein peaked after 5.5 and 7.4 h, respectively. The systemic bioavailability and maximum serum concentration of [(13)C]genistein were significantly greater than those of [(13)C]daidzein. The bioavailability of both isoflavones did not increase linearly when the dietary intake was doubled. The mean volume of distribution normalized to bioavailability (V(d)/F), clearance rate, and half-life of [(13)C]daidzein were 336.25 L, 30.09 L/h, and 7.75 h, respectively; the corresponding values for [(13)C]genistein were 258.76 L, 21.85 L/h, and 7.77 h. The average recovery of [(13)C]daidzein and [(13)C]genistein in urine was 30.1% and 9.0% of the dose ingested, respectively. CONCLUSIONS: The serum pharmacokinetics of [(13)C]daidzein and [(13)C]genistein were reproducible among healthy women, and genistein was more bioavailable than was daidzein. Pharmacokinetics were unaffected by chronic exposure to soy foods. Urinary isoflavone concentrations correlated poorly with maximal serum concentrations, indicating the limitations of urine measurements as a predictor of systemic bioavailability. The bioavailability of both isoflavones was nonlinear at higher intakes, suggesting that uptake is rate-limiting and saturable. PMID- 12540403 TI - Vitamin B-12 metabolism in HIV-infected patients in the age of highly active antiretroviral therapy: role of homocysteine in assessing vitamin B-12 status. AB - BACKGROUND: Before the advent of highly active antiretroviral therapy (HAART), 20% and 10% of HIV-infected patients had low vitamin B-12 and red blood cell folate (RBCF) concentrations, respectively. However, few patients had real vitamin B-12 deficiency. OBJECTIVE: We evaluated the prevalence of low vitamin B 12 and RBCF concentrations in HIV-infected patients receiving HAART and the usefulness of serum homocysteine (sHcy) for differentiating patients with deficiency from those with harmlessly low vitamin B-12. DESIGN: The prevalence of low vitamin B-12 and RBCF was evaluated in 126 HIV-infected patients receiving HAART. Moreover, sHcy concentrations were evaluated in 40 HIV-infected patients with low vitamin B-12 and in 37 HIV-infected patients with low RBCF and were compared with those in 128 HIV-infected patients with normal vitamin B-12 and RBCF. sHcy was used to monitor treatment with vitamin B-12 and folic acid in 28 patients (24 with low vitamin B-12 and RBCF and 4 with hyperhomocysteinemia but normal vitamin B-12 and RBCF). RESULTS: The prevalence of low vitamin B-12 was significantly lower in patients receiving HAART than in previously studied patients who did not receive HAART (8.7% compared with 27%). Nine of the 40 patients (22.5%) with low vitamin B-12 (< or = 200 pmol/L) had hyperhomocysteinemia (> 17.5 micromol homocysteine/L). Nineteen (51.4%) of the 37 patients with low RBCF (< or = 580 nmol/L, percentile 10) had hyperhomocysteinemia. Among the 9 patients with an RBCF concentration < or = 450 nmol/L (percentile 2.5), all had hyperhomocysteinemia. The treatment with vitamin B-12 and folic acid normalized sHcy concentrations. CONCLUSIONS: The prevalence of low vitamin B-12 decreased after the introduction of HAART. The study of sHcy is useful for detecting HIV-infected patients with low vitamin B-12 and real deficiency. PMID- 12540404 TI - Dual fortification of salt with iodine and microencapsulated iron: a randomized, double-blind, controlled trial in Moroccan schoolchildren. AB - BACKGROUND: In many developing countries, children are at high risk of both goiter and iron deficiency anemia. OBJECTIVE: In a series of studies in northern Morocco, we developed and tested a dual-fortified salt (DFS) containing iodine and microencapsulated iron. DESIGN: To establish the DFS fortification concentration, we measured salt intake by 3-d weighed food records and estimated iron bioavailability from the local diet by using published algorithms. We then formulated a DFS containing 25 micro g iodine/g salt (as potassium iodide) and 1 mg iron/g salt (as ferrous sulfate hydrate encapsulated with partially hydrogenated vegetable oil). After storage and acceptability trials, we compared the efficacy of the DFS to that of iodized salt in a 9-mo, randomized, double blind trial in iodine-deficient, 6-15-y-old children (n = 377). RESULTS: Mean salt intake in school-age children was 7-12 g/d, and estimated iron bioavailability from the local diet was 0.4-4.3%. After storage for 20 wk, the DFS and iodized salt were not significantly different in iodine content, and color stability was acceptable when the compounds were added to local meals. During the efficacy trial, urinary iodine concentrations and thyroid volumes improved significantly (P < 0.001 and < 0.05, respectively) from baseline in both groups. At 40 wk, mean hemoglobin concentrations in the DFS group had increased by 14 g/L (P < 0.01), and serum ferritin, transferrin receptor, and zinc protoporphyrin concentrations were significantly better (P < 0.05) in the DFS group than in the iodized salt group. The prevalence of iron deficiency anemia in the DFS group decreased from 35% at baseline to 8% at 40 wk (P < 0.001). CONCLUSION: A DFS containing iodine and encapsulated iron can be an effective fortification strategy. PMID- 12540405 TI - High prevalence of cobalamin deficiency in Guatemalan schoolchildren: associations with low plasma holotranscobalamin II and elevated serum methylmalonic acid and plasma homocysteine concentrations. AB - BACKGROUND: Studies conducted in Guatemala, Mexico, and Venezuela have found high prevalences of low plasma cobalamin (vitamin B-12) concentrations in infants and children. It is not known whether these low cobalamin concentrations are accompanied by altered metabolic functions. OBJECTIVE: We sought to assess the prevalence of cobalamin deficiency in Guatemalan children by using sensitive and specific markers of deficiency. DESIGN: Children (n = 553) were screened for low plasma cobalamin. Those with low plasma cobalamin (< 162 pmol/L) were matched by age, grade, and sex to those with marginal (162-221 pmol/L) and adequate (> 221 pmol/L) concentrations. In this matched subset (n = 180), additional biochemical indicators of cobalamin deficiency were measured. RESULTS: Of the 553 children screened, 11% had low plasma cobalamin and an additional 22% had marginal concentrations. The prevalences of elevated serum methylmalonic acid (MMA), plasma homocysteine, or both were significantly higher in children with low and marginal plasma cobalamin than in children with adequate plasma cobalamin. Mean serum MMA was high in all groups compared with values reported in other populations. Mean plasma holotranscobalamin II concentrations were significantly lower in children with low rather than marginal or adequate plasma cobalamin. However, holotranscobalamin II was a less sensitive indicator of cobalamin depletion than was MMA. CONCLUSION: Biochemical markers of cobalamin deficiency confirmed that the cobalamin status of children with low and marginal plasma cobalamin is inadequate to support normal metabolic function. PMID- 12540406 TI - Iron supplementation improves progressive fatigue resistance during dynamic knee extensor exercise in iron-depleted, nonanemic women. AB - BACKGROUND: Tissue iron depletion may negatively affect endurance performance and muscle fatigability. OBJECTIVE: We investigated tissue-level iron depletion and progressive fatigue of the quadriceps during dynamic knee-extension exercise in young women. DESIGN: Twenty iron-depleted (serum ferritin < 20 micro g/L), nonanemic (hemoglobin > 110 g/L) women (macro x +/- SEM age: 29.1 +/- 1.2 y) received iron (iron group) or placebo (placebo group) for 6 wk in a randomized, double-blind trial (n = 10 per group). A protocol integrating 2-3-s maximal voluntary static contractions (MVCs) with dynamic knee extensions was used to assess fatigue. RESULTS: No significant differences between the groups in baseline iron status, MVC at rest, or MVC at the end of the protocol were observed. After treatment, serum iron and transferrin saturation increased significantly in the iron group (P = 0.02 and P = 0.03, respectively). Serum transferrin receptor concentrations increased significantly in the placebo group (P < 0.01) but not in the iron group. After treatment, the rate of decrease in MVC was attenuated in the iron group but not in the placebo group (P = 0.01). In the iron group, MVC at the sixth minute of the fatigue protocol and MVC at the end of the protocol were approximately 15% (P = 0.04) and approximately 27% higher (P < 0.01), respectively, after treatment. These improvements were not related to changes in iron-status indexes or tissue iron stores, although power was low (< 0.50) to detect these relations. CONCLUSIONS: Iron supplementation was associated with a significant improvement in muscle fatigability. Interpretation regarding the direct role of tissue iron status is limited by the study's low power to detect relations between tissue iron improvement and decreased muscle fatigue. PMID- 12540407 TI - Effect of short-chain fructooligosaccharides on intestinal calcium absorption and calcium status in postmenopausal women: a stable-isotope study. AB - BACKGROUND: An enhancing effect of short-chain fructooligosaccharides (scFOSs) on intestinal calcium absorption has been shown in animals and in some short-term human studies. However, the long-term effect of scFOSs on calcium absorption in humans is still unknown. OBJECTIVE: We investigated the long-term effect of a moderate daily dose (10 g) of scFOSs on intestinal calcium absorption in postmenopausal women. DESIGN: In a randomized, double-blind crossover protocol, 12 healthy, postmenopausal women received scFOSs or placebo for 5 wk. The treatments were separated by a 3-wk washout period. Subjects orally received (44)Ca (stable isotope) and a fecal marker. Feces were collected after the isotope intake for 5-7 d to measure unabsorbed isotope. Calcium-status indexes, calciotropic hormones, and bone turnover were also assessed. RESULTS: Mean (+/ SD) intestinal calcium absorption with scFOS treatment was not significantly different from that with placebo treatment (35.63 +/- 9.40% and 36.55 +/- 8.48%, respectively). However, a tendency for calcium absorption to be higher with scFOS treatment than with placebo treatment was observed in women who had been going through menopause for >6 y. CONCLUSIONS: scFOSs do not modify intestinal calcium absorption in postmenopausal women who do not receive hormonal replacement therapy. The results from a subgroup of women who had been going through menopause for >6 y (n = 6) suggest that scFOSs may influence calcium absorption in the late postmenopausal phase. The small number of subjects and the related P value warrant verification and further investigation with women in late menopause only. PMID- 12540408 TI - Lipid standardization of serum fat-soluble antioxidant concentrations: the YALTA study. AB - BACKGROUND: Blood lipids can influence fat-soluble antioxidant concentrations and confound their interpretation as indicators of antioxidant intake status and disease risk. OBJECTIVES: The objectives were to identify lipoproteins that can confound the interpretation of serum fat-soluble antioxidants, to evaluate the amount of the confounding, and to recommend a method for standardizing blood concentrations of fat-soluble antioxidants. DESIGN: Several methods of lipid standardization of fat-soluble antioxidants were evaluated in a large cohort of young adults with the use of both cross-sectional and longitudinal data analysis. RESULTS: Tocopherol and carotenoid concentrations were associated with plasma total cholesterol and its components, LDL, HDL, and VLDL cholesterol (estimated as plasma total triacylglycerols/5), some of which were independent predictors for all of the fat-soluble antioxidants. Among supplement nonusers, the most amphipathic (polar) of the antioxidants (alpha-tocopherol, gamma-tocopherol, and zeaxanthin plus lutein) and lycopene were associated strongly with these lipid fractions (R(2) = 0.09, 0.40). Consistent with a causal association in which blood antioxidant concentrations change as blood lipid concentrations change, similar relations were found for changes in blood antioxidant and lipid concentrations over a 7-y period. Concentrations of the remaining carotenoids (beta-cryptoxanthin, alpha-carotene, and beta-carotene) had a weaker association with plasma lipoproteins (R(2) < 0.06). Similar relations were found for supplement users. CONCLUSIONS: The simultaneous adjustment of the concentrations of tocopherols, zeaxanthin plus lutein, and lycopene for VLDL, HDL, and LDL cholesterol is recommended. This method is practical and can provide a basis for the standardization of carotenoid and tocopherol concentrations. PMID- 12540409 TI - Plasma total cysteine, pregnancy complications, and adverse pregnancy outcomes: the Hordaland Homocysteine Study. AB - BACKGROUND: Total homocysteine (tHcy) is associated with pregnancy complications and adverse pregnancy outcomes. The associations of plasma total cysteine (tCys) with such outcomes have not been investigated in large populations. OBJECTIVE: We investigated the association between plasma tCys and pregnancy complications, congenital malformations, and other adverse pregnancy outcomes. DESIGN: The plasma tCys concentrations of 5883 women aged 40-42 y that were measured in 1992 1993 during a cardiovascular health screening were compared with the outcomes and complications of 14492 pregnancies in the same women that were registered in the Medical Birth Registry of Norway from 1967 to 1996. RESULTS: After adjustment for parity, mother's age, tHcy, total cholesterol, body mass index, smoking, and coffee drinking, high plasma tCys concentrations (above the 95th percentile) were associated with significantly higher risks of preeclampsia [n = 342; odds ratio (OR): 1.6; 95% CI: 1.1, 2.4; P = 0.03], premature delivery (n = 774; OR: 1.8; 95% CI: 1.3, 2.5; P = 0.001), and very low birth weight (n = 175; OR: 2.0; 95% CI: 1.1, 3.9; P = 0.03) than were lower plasma tCys concentrations. tCys was not associated with the risk of placental abruption. High tCys concentrations showed a weak association with congenital malformations and stillbirths with birth weight <1500 g. The associations were independent of the tHcy concentrations. CONCLUSION: High tCys concentrations were associated with risks of preeclampsia, premature delivery, and low birth weight. PMID- 12540410 TI - Intakes of essential n-6 and n-3 polyunsaturated fatty acids among pregnant Canadian women. AB - BACKGROUND: Fetal growth requires n-3 docosahexaenoic acid (DHA), which is derived from the essential n-3 fatty acids in the maternal diet. DHA is accumulated in the developing brain and is critical for normal neural and visual function. Available estimates suggest that 67 mg DHA/d is accumulated by the fetus during the third trimester of gestation. Little is known about n-3 fatty acid intakes in pregnant women, although human milk concentrations of DHA have decreased in recent years. OBJECTIVE: We prospectively determined the n-3 and n-6 fatty acid intakes of 55 pregnant Canadian women. DESIGN: A food-frequency questionnaire was completed at 28 and 35 wk, and plasma n-3 and n-6 fatty acids were measured at 35 wk gestation. The fatty acid composition of approximately 500 foods was analyzed to allow analysis of dietary intakes from specific foods. RESULTS: Intakes, as a percentage of energy, were (macro x +/- SEM) total fat, 28.0 +/- 3.6%; saturated fat, 9.8 +/- 0.3%; monounsaturated fat, 11.2 +/- 0.4%; polyunsaturated fat, 4.7 +/- 0.2%; linoleic acid, 3.9 +/- 0.2%; and alpha linolenic acid, 0.54 +/- 0.05%. The daily intakes (range) were 160 +/- 20 (24 524) mg DHA/d, 121 +/- 8 (15-301) mg arachidonic acid/d, and 78 +/- 2 (4-125) mg eicosapentaenoic acid/d. The plasma phospholipids had (mg/100 g fatty acid) 5.0 +/- 0.18 DHA, 8.7 +/- 0.18 arachidonic acid, and 0.52 +/- 0.32 eicosapentaenoic acid. CONCLUSION: The low intake of DHA among some pregnant women highlights the need for studies to address the functional significance of maternal fat intakes during pregnancy on fetal development. PMID- 12540411 TI - Metabolic adaptation to high-fat and high-carbohydrate diets in children and adolescents. AB - BACKGROUND: Difficulty adapting to high-fat (HF) and high carbohydrate (HC) diets may predispose children to obesity and diabetes. OBJECTIVE: We tested the hypothesis that children have metabolic flexibility to adapt to HF and HC diets. DESIGN: In protocol 1, 12 children aged 6-9 y and 12 adolescents aged 13-16 y were randomly assigned in a crossover design to consume low-fat (LF), HC (25% and 60% of energy, respectively) or HF, low-carbohydrate (LC) (55% and 30% of energy, respectively) diets. In protocol 2, 12 adolescents aged 13-16 y were randomly assigned in a crossover design to consume an LF-HC diet with 11% or 40% of carbohydrate as fructose. Total energy expenditure, nonprotein respiratory quotients (NPRQs), and substrate utilization were measured by using 24-h calorimetry. Effects of sex, puberty, body fat (dual-energy X-ray absorptiometry), intraabdominal fat (magnetic resonance imaging), and fitness on substrate utilization were tested. RESULTS: Substrate utilization was not affected by puberty, body fat, intraabdominal fat, or fitness. Total energy expenditure was not affected by diet. In protocol 1, NPRQs and carbohydrate and fat utilization were significantly affected by diet (P = 0.001) and sex (P = 0.005). NPRQs and carbohydrate utilization increased with the LF-HC diet. NPRQs decreased and fat utilization increased with the HF-LC diet; changes in substrate utilization were less pronounced in females than in males. In protocol 2, 24-h NPRQs and 24-h substrate utilization were not significantly affected by fructose, although net carbohydrate and fat utilization were significantly lower and higher, respectively, with the high-fructose diet during fasting (P = 0.01) and in the subsequent feeding period (P = 0.05). CONCLUSION: Healthy, nonobese children and adolescents adapt appropriately to HF and HC diets. PMID- 12540412 TI - Inhibition of lipolysis improves insulin sensitivity in protease inhibitor treated HIV-infected men with fat redistribution. AB - BACKGROUND: Fatty acid concentrations are increased in patients with HIV and fat redistribution and may contribute to insulin resistance in this population. OBJECTIVE: We determined the effects of acute inhibition of lipolysis on insulin sensitivity in HIV-infected patients with fat redistribution who were receiving a protease inhibitor. DESIGN: Seven HIV-infected men [age: 45 +/- 2 y; body mass index (in kg/m(2)): 28.8 +/- 1.9] with a fasting insulin concentration > or= 104 pmol/L (15 micro IU/mL), combined visceral adiposity and peripheral lipoatrophy, and receiving a protease inhibitor were studied. Tolbutamide-modified frequently sampled intravenous-glucose-tolerance tests (FSIGTTs) were performed after randomized double-blind administration of acipimox (500 mg at -90 and 0 min), a potent inhibitor of lipolysis, and placebo. The subjects completed 2 FSIGTTs separated by 3-7 d. RESULTS: At baseline, fasting insulin and fatty acid concentrations were 27.6 +/- 5.0 micro IU/mL and 0.83 +/- 0.08 mmol/L (normal range: 0.1-0.6 mmol/L), respectively. Fatty acid concentrations were significantly reduced after acipimox compared with placebo (fatty acid area under the curve: acipimox = 73 +/- 8 compared with placebo = 122 +/- 12 mmol x 270 min/L, P = 0.002). Acipimox treatment resulted in a significant increase in the insulin sensitivity index (acipimox = 1.63 +/- 0.5 compared with placebo = 0.88 +/- 0.3 x 10(-4) x min(-1) x micro IU/mL, P = 0.015). CONCLUSIONS: Acute inhibition of lipolysis and reduction in fatty acid concentrations are associated with improved insulin sensitivity in patients with HIV lipodystrophy and hyperinsulinemia. Further studies are needed to determine whether long-term antilipolytic strategies to reduce fatty acid concentrations may be useful in treating the metabolic disturbances associated with HIV lipodystrophy. PMID- 12540413 TI - Diet in midpuberty and sedentary activity in prepuberty predict peak bone mass. AB - BACKGROUND: An average daily calcium intake of 1300 mg is recommended for North American adolescents aged 9-18 y. However, questions remain about these recommendations. OBJECTIVE: We assessed whether there is a stage of puberty when dietary calcium is more strongly related to peak bone mass, as indicated by young adult bone mass (YABM); whether dietary calcium intake > 1000 mg/d in adolescence is associated with higher YABM; and whether race affects any of these associations between dietary calcium and YABM. Secondarily, we evaluated relations between sedentariness and YABM. DESIGN: In a retrospective cohort study, we recruited 693 black and white women aged 21-24 y who had participated in the 10-y National Heart, Lung, and Blood Institute Growth and Health Study and measured YABM with the use of dual-energy X-ray absorptiometry. Dietary calcium and sedentary activity data, gathered through 3-d food records and self-reports of television-video viewing at 8 annual examinations, were averaged over 3 pubertal stages. Complete data were available from 161 black and 180 white females. Multiple regression, controlling for race, weight, and height, was applied to assess diet and activity relations with YABM. RESULTS: Dietary calcium was most strongly associated with YABM in midpuberty. Calcium intake > 1000 mg/d was associated with higher YABM, but this association was not significant at all skeletal sites. Race did not affect the observed relations between calcium and YABM. Sedentary activity in prepuberty was inversely associated with YABM. CONCLUSIONS: Interventions should focus on ensuring adequate calcium intake in midpuberty and on minimizing sedentariness in prepuberty. PMID- 12540414 TI - Calcium, vitamin D, milk consumption, and hip fractures: a prospective study among postmenopausal women. AB - BACKGROUND: Short trials of calcium supplementation show that it reduces loss of bone density in postmenopausal women; longer observational studies do not generally find a lower risk of hip fracture with higher-calcium diets. Fewer studies have focused on vitamin D in preventing postmenopausal osteoporosis or fractures. OBJECTIVE: We assessed relations between postmenopausal hip fracture risk and calcium, vitamin D, and milk consumption. DESIGN: In an 18-y prospective analysis in 72 337 postmenopausal women, dietary intake and nutritional supplement use were assessed at baseline in 1980 and updated several times during follow-up. We identified 603 incident hip fractures resulting from low or moderate trauma. Relative risks (RRs) from proportional hazards models were controlled for other dietary and nondietary factors. RESULTS: Women consuming > or = 12.5 microg vitamin D/d from food plus supplements had a 37% lower risk of hip fracture (RR = 0.63; 95% CI: 0.42, 0.94) than did women consuming < 3.5 microg/d. Total calcium intake was not associated with hip fracture risk (RR = 0.96; 95% CI: 0.68, 1.34 for > or = 1200 compared with < 600 mg/d). Milk consumption was also not associated with a lower risk of hip fracture (P for trend = 0.21). CONCLUSIONS: An adequate vitamin D intake is associated with a lower risk of osteoporotic hip fractures in postmenopausal women. Neither milk nor a high-calcium diet appears to reduce risk. Because women commonly consume less than the recommended intake of vitamin D, supplement use or dark fish consumption may be prudent. PMID- 12540415 TI - Vitamin K intake and bone mineral density in women and men. AB - BACKGROUND: Low dietary vitamin K intake has been associated with an increased risk of hip fracture in men and women. Few data exist on the association between dietary vitamin K intake and bone mineral density (BMD). OBJECTIVE: We studied cross-sectional associations between self-reported dietary vitamin K intake and BMD of the hip and spine in men and women aged 29-86 y. DESIGN: BMD was measured at the hip and spine in 1112 men and 1479 women (macro x +/- SD age: 59 +/- 9 y) who participated in the Framingham Heart Study (1996-2000). Dietary and supplemental intakes of vitamin K were assessed with the use of a food-frequency questionnaire. Additional covariates included age, body mass index, smoking status, alcohol use, physical activity score, and menopause status and current estrogen use among the women. RESULTS: Women in the lowest quartile of vitamin K intake (macro x: 70.2 microg/d) had significantly (P < or = 0.005) lower mean (+/ SEM) BMD at the femoral neck (0.854 +/- 0.006 g/cm(2)) and spine (1.140 +/- 0.010 g/cm(2)) than did those in the highest quartile of vitamin K intake (macro x: 309 microg/d): 0.888 +/- 0.006 and 1.190 +/- 0.010 g/cm(2), respectively. These associations remained after potential confounders were controlled for and after stratification by age or supplement use. No significant association was found between dietary vitamin K intake and BMD in men. CONCLUSIONS: Low dietary vitamin K intake was associated with low BMD in women, consistent with previous reports that low dietary vitamin K intake is associated with an increased risk of hip fracture. In contrast, there was no association between dietary vitamin K intake and BMD in men. PMID- 12540417 TI - Alpha-linolenic acid, linoleic acid, coronary artery disease, and overall mortality. PMID- 12540416 TI - Ingested probiotics reduce nasal colonization with pathogenic bacteria (Staphylococcus aureus, Streptococcus pneumoniae, and beta-hemolytic streptococci). AB - BACKGROUND: As a bacterial reservoir, the nose may harbor potentially pathogenic bacteria (PPB: Staphylococcus aureus, Streptococcus pneumoniae, beta-hemolytic streptococci, and Haemophilus influenzae). In patients carrying PPB, antiseptic regimens could be crucial for infection control after major operations on or injuries of the head, nasal sinuses, or lungs. Such regimens may also be important for diabetic patients and persons receiving hemodialysis, in intensive care units, or with impaired immunity due to various other causes. OBJECTIVE: We tested a possible effect of the ingestion of probiotics on the bacterial flora of the nose. DESIGN: In an open, prospective trial, 209 volunteers were randomly assigned to consume either a probiotic, fermented milk drink [65 mL with Lactobacillus GG (ATCC 53103), Bifidobacterium sp B420, Lactobacillus acidophilus 145, and Streptococcus thermophilus; n = 108] or standard yogurt (180 g; n = 101) daily for 3 wk. Nasal microbial flora were analyzed on days 1, 21, and 28. The microbial examination was blinded to the source of the samples. RESULTS: We found a significant reduction (19%; P < 0.001) in the occurrence of nasal PPB in the group who consumed the probiotic drink but not in the group who consumed yogurt. The effect was mainly on gram-positive bacteria, which decreased significantly (P < 0.05). CONCLUSIONS: The results indicate that regular intake of probiotics can reduce PPB in the upper respiratory tract. The results also indicate a linkage of the lymphoid tissue between the gut and the upper respiratory tract. PMID- 12540418 TI - Issues of standardization and assay-specific clinical decision limits for the measurement of 25-hydroxyvitamin D. PMID- 12540419 TI - Low-dose CT: how are we doing? PMID- 12540422 TI - Can low-dose unenhanced multidetector CT be used for routine evaluation of suspected renal colic? PMID- 12540420 TI - Low-dose unenhanced multidetector CT of patients with suspected renal colic. AB - OBJECTIVE: This study is designed to assess the intraobserver and interobserver agreements and the diagnostic performances of low-dose unenhanced multidetector CT (MDCT) in patients with suspected renal colic. SUBJECTS AND METHODS: The study included 106 patients who underwent unenhanced MDCT with 4 x 2.5 mm collimation, 120 kVp, 30 mAs, and, if necessary, additional focused acquisitions at 60 or 120 mAs on areas with an equivocal ureteral stone or with significant image noise. The effective radiation dose was computer-simulated with software based on the Monte Carlo model and International Commission on Radiological Protection recommendations. CT scans were archived and independently reviewed by three radiologists during two interpretation sessions on a workstation with three dimensions functions. Intraobserver and interobserver agreements were calculated with the kappa statistics. Accuracy for detection of ureteral stone on low-dose MDCT was calculated by comparison with combined clinical (stone passage), surgical (stone retrieval, extracorporeal shock wave lithotripsy), biologic (urinalysis, urine culture), and other imaging (excretory urography, standard dose MDCT, follow-up sonography, and abdominal radiography) findings or by evidence for an alternative diagnosis. RESULTS: Ureteral stones were present in 38 (36%) of 106 patients. Thirty-six of 38 ureteral stones were detected by low dose MDCT. From reviewer to reviewer, the number of true-positive, false positive, true-negative, and false-negative findings ranged, respectively, from 34 to 36, 1 to 4, 64 to 68, and 2 to 4. The corresponding sensitivity, specificity, and accuracy ranged from 89.5% to 94.7%, from 94.1% to 100%, and from 93.4% to 98.1%, respectively. The intraobserver and interobserver agreements were excellent, with kappa values ranging from 0.87 to 0.98. In 13 patients, an alternative diagnosis explaining the patient's symptoms was proposed by all reviewers using images obtained at 30 mAs. No additional or alternative diagnosis was found at standard dose. At 30 mAs, the mean effective dose was 1.2 mSv in men and 1.9 mSv in women. Additional acquisitions at 60 mAs, all focused on the lower pelvis, were acquired in 20 patients, but the corresponding images were needed by the reviewers for only six of them. The acquisitions at 60 mAs were responsible for an additional mean effective dose of 0.5 in men and 0.8 mSv in women. CONCLUSION: Our study shows that low-dose unenhanced MDCT is appropriate for the diagnosis of ureteral stones, and that it provides excellent intraobserver and interobserver agreements and does not obscure alternative diagnoses. PMID- 12540423 TI - Potential legal ramifications of whole-body CT screening: taking a peek into Pandora's box. PMID- 12540424 TI - Should whole-body CT screening be performed with contrast media? PMID- 12540425 TI - Style guidelines for radiology reporting: a manner of speaking. PMID- 12540426 TI - Improving image contrast. PMID- 12540427 TI - Radiologic-pathologic conferences of the University of Texas M.D. Anderson Cancer Center. Phyllodes tumor of the breast. PMID- 12540428 TI - MR imaging findings in the contralateral breast of women with recently diagnosed breast cancer. AB - OBJECTIVE: The purpose of this study was to determine the frequency and positive predictive value of biopsy performed on the basis of MR imaging findings in the contralateral breast in women with recently diagnosed breast cancer. MATERIALS AND METHODS: We performed a retrospective review of records of 1336 consecutive breast MR imaging examinations over a 2-year period. Of these examinations, 223 imaged the asymptomatic, mammographically normal contralateral breast in women whose breast cancer was diagnosed within 6 months preceding MR imaging. Records of these 223 examinations were reviewed to determine the frequency of recommending contralateral breast biopsy and the biopsy results. RESULTS: Contralateral breast biopsy was recommended in 72 (32%) of 223 women and performed in 61 women. Cancer occult to mammography and physical examination was detected by MR imaging in 12 women, constituting 20% (12/61) of women who underwent contralateral biopsy and 5% (12/223) of women who underwent contralateral breast MR imaging. Among these 12 cancers, six (50%) were ductal carcinoma in situ (DCIS) and six (50%) were infiltrating carcinoma. The median size of infiltrating carcinoma was 0.5 cm (range, 0.1-1.0 cm). Contralateral biopsy revealed benign (n = 31) or high-risk (n = 18) lesions in 49 women, constituting 80% (49/61) of women who underwent contralateral biopsy and 22% (49/223) of women who underwent contralateral MR imaging. CONCLUSION: In women with recently diagnosed breast cancer, MR imaging of the contralateral breast led to a biopsy recommendation in 32%. Cancer was found in 20% of women who underwent contralateral breast biopsy and in 5% of women who underwent contralateral breast MR imaging. PMID- 12540430 TI - Optimal reference mammography: a comparison of mammograms obtained 1 and 2 years before the present examination. AB - OBJECTIVE: We assessed and compared the benefit of using images acquired 1 year or 2 years previously during mammography interpretations. MATERIALS AND METHODS: Eleven radiologists and one resident reviewed 128 cases three times: once without prior mammograms for comparison, once with mammograms from the most recent (1 year) examination, and once with mammograms acquired 2 years previously. They were asked to determine whether the patient should be recalled for additional procedures. Performances under the three conditions were compared. RESULTS: Radiologists were significantly more accurate (p < 0.001) when comparison mammograms (obtained 1 or 2 years previously) were available. Although sensitivity was not significantly affected between the availability of mammograms from 1 or 2 years earlier (p > 0.10), the specificity was. Specificity using mammograms from the latest examination (obtained 1 year previously) as a reference was significantly better (p = 0.03) than specificity using mammograms obtained 2 years previously. CONCLUSION: Comparison mammograms are important for accurate diagnosis-in particular, for increasing specificity. The latest prior examination seems to be the optimal one for this purpose. PMID- 12540431 TI - Comparison of automated versus vacuum-assisted biopsy methods for sonographically guided core biopsy of the breast. AB - OBJECTIVE: The purpose of this study was to compare the outcome of sonographically guided core biopsies performed with the 14-gauge automated gun with the outcome of those performed with the 11-gauge vacuum-assisted device. Outcome was defined in terms of missed tumors, the need (both immediate and delayed) for a second biopsy, histologic underestimation, and complication rates. MATERIALS AND METHODS: We retrospectively reviewed all sonographically guided core biopsies performed between January 1997 and August 2001. Before February 2000, biopsies were performed using the 14-gauge automated gun and after that time, with either the 14-gauge automated gun or the 11-gauge vacuum-assisted device. During the study period, 181 biopsies were performed with the 14-gauge automated gun and 100 with the 11-gauge vacuum-assisted device. RESULTS: The histologic results of the core biopsies were similar for the group who underwent biopsy with the 14-gauge automated gun and the group who underwent biopsy with the 11-gauge vacuum-assisted device: malignant, 19% versus 19%; benign, 78% versus 79%; and high-risk lesion or other, 3% versus 2%, respectively (p > 0.7). Complications were rare and similar for both methods: 2% for the 14-gauge automated gun and 3% for the 11-gauge vacuum-assisted device (p = 0.46). A second biopsy was recommended immediately after the first in 14% of the patients who underwent biopsy with the 14-gauge automated gun versus 17% of those who underwent biopsy with the 11-gauge vacuum-assisted device (p = 0.47). Recommendation for delayed rebiopsy due to interval change occurred in 2.5% of the patients who underwent biopsy with the 14-gauge automated gun method and 3% of those who underwent biopsy with the 11-gauge vacuum-assisted device (p = 0.94). CONCLUSION: No significant differences were found in the outcomes of sonographically guided core biopsies performed with the automated gun compared with those performed with the vacuum-assisted device in terms of missed cancers, underestimation, complications, or the need (immediate or delayed) for a second biopsy. PMID- 12540432 TI - Sonographically guided needle localization after stereotactic breast biopsy. PMID- 12540433 TI - Marking the cavity site after stereotactic core needle breast biopsy. PMID- 12540434 TI - Rapid MR imaging of articular cartilage with steady-state free precession and multipoint fat-water separation. AB - OBJECTIVE: To obtain high-quality high-resolution images of articular cartilage with reduced imaging time, we combined a novel technique of generalized multipoint fat-water separation with three-dimensional (3D) steady-state free precession (SSFP) imaging. SUBJECTS AND METHODS: The cartilage of 10 knees in five healthy volunteers was imaged with 3D SSFP imaging and a multipoint fat water separation method capable of separating fat and water with short TE increments. Fat-saturated 3D spoiled gradient-echo (SPGR) images were obtained for comparison. RESULTS: High-quality images of the knee with excellent fat-water separation were obtained with 3D SSFP imaging. Total imaging time required was 58% less than that required for 3D SPGR imaging with a comparable cartilage signal-to-noise ratio and spatial resolution. Unlike 3D SPGR images, 3D SSFP images exhibited bright synovial fluid, providing a potential arthrographic effect. CONCLUSION: High-quality high-resolution images of articular cartilage with improved fat-water separation, bright synovial fluid, and markedly reduced acquisition times can be obtained with 3D SSFP imaging combined with a fat-water separation technique. PMID- 12540435 TI - Subchondral fractures in osteonecrosis of the femoral head: comparison of radiography, CT, and MR imaging. AB - OBJECTIVE: Our objective was to compare the sensitivity of unenhanced radiography, CT, and MR imaging in revealing subchondral fractures. SUBJECTS AND METHODS: Forty-five subjects with stage I and stage II osteonecrosis of the femoral head were included in the study as part of a multicenter clinical trial to evaluate the effectiveness of recombinant human bone morphogenetic protein as an adjuvant treatment to core decompression. Patients were evaluated with radiography, CT, and MR imaging 6 and 12 months after surgery. RESULTS: At 6 months, 18 fractures were shown on CT scans, but only 12 were detected on radiographs and six, on MR images. At 12 months, 20 subchondral fractures were detected on CT scans, but only 17 were seen on radiographs and 11, on MR images. Compared with CT, MR imaging has a sensitivity and specificity of 38% and 100%, and unenhanced radiography has a sensitivity and specificity of 71% and 97%, respectively. On T2-weighted MR images, the subchondral fractures were visualized as crescentic high-signal-intensity lines, and in all patients, on the corresponding CT scans, the fracture clearly breached the femoral cortex. CONCLUSION: CT reveals more subchondral fractures in osteonecrosis of the femoral head than unenhanced radiography or MR imaging. The high-signal-intensity line seen on T2-weighted MR images appears to represent fluid accumulating in the subchondral fracture, which may indicate a breach in the overlying articular cartilage. PMID- 12540436 TI - MR arthrography of the posterior labrocapsular complex: relationship with glenohumeral joint alignment and clinical posterior instability. AB - OBJECTIVE: The purpose of our study was to investigate the relationship between tears of the posterior labrocapsular complex and glenohumeral alignment on MR arthrography and the presence and extent of posterior labrocapsular tears in patients with posterior instability. MATERIALS AND METHODS: Posterior labrocapsular tears identified on 24 MR arthrograms and surgically confirmed were evaluated for length of tear and labrocapsular avulsion. These examinations and a comparison cohort of 70 normal MR arthrograms with normal findings were also evaluated for humeral head position relative to the glenoid fossa. Medical records were reviewed for clinical diagnosis of posterior instability and history of shoulder trauma. RESULTS: The position of the humeral head relative to the glenoid was significantly more posterior in patients with posterior labral tear than in patients with a normal posterior labrum (4.9 mm versus 0.7 mm; p < 0.0001). The mean length (+/- SD) of posterior labral tear was 15.9 +/- 1.7 mm, and a direct correlation was found between tear length and posterior humeral translation (r = -0.65; p = 0.002). Posterior labral tears were significantly longer (18.6 vs 13.1 mm; p = 0.04), and posterior humeral translation was greater (6.4 vs 3.4 mm; p = 0.006) in patients with labrocapsular avulsion than in those without avulsion. Twelve (50%) of the patients with posterior labrocapsular tear had posterior instability, and 10 (83%) had a history of macrotrauma. On MR arthrography, the mean posterior humeral translation was greater (6.2 mm +/- 0.08; p = 0.019), posterior labral tears were longer (19.4 mm +/- 1.7; p = 0.0008), and labrocapsular avulsion was more common (83%; p = 0.0001) in patients with posterior instability than in patients who had a posterior labral tear but a clinically stable shoulder. CONCLUSION: Clinical posterior instability is associated with excessive posterior humeral translation, long posterior labral tears, and posterior labrocapsular avulsion. PMID- 12540437 TI - MR imaging of complications of loose surgical tacks in the shoulder. AB - OBJECTIVE: Loose surgical tacks in the shoulder joint are a potential cause of new-onset shoulder pain after arthroscopic repair of an anterior-to-posterior lesion of the superior labrum. We report the MR imaging appearance of loose surgical tacks in this anatomic location. CONCLUSION: MR imaging is valuable in the evaluation of postoperative shoulder pain. Synovitis is a commonly considered clinical diagnosis; our report illustrates that loose tacks are another potential complication after shoulder surgery PMID- 12540438 TI - Avulsion fracture of the head of the fibula (the "arcuate" sign): MR imaging findings predictive of injuries to the posterolateral ligaments and posterior cruciate ligament. AB - OBJECTIVE: The purpose of this study was to describe the significance of an avulsion fracture of the head of the fibula ("arcuate" sign) and its association with injuries of the knee on MR imaging. MATERIALS AND METHODS: We conducted a retrospective search of 2318 patients who underwent conventional radiography and MR imaging after an episode of knee trauma. Patients were included in this study if they had an avulsion fracture of the head of the fibula revealed on conventional radiography and underwent arthroscopy. Thirteen patients, all of whom were men, satisfied the inclusion criteria. Ten patients underwent further explorative surgery. The clinical, radiographic, MR imaging, and surgical findings were then reviewed. RESULTS: The avulsion fracture of the styloid process of the fibular head was apparently related to injuries of the arcuate complex in all 13 patients. Radiographically, the bony fragment was horizontally oriented and similar in size in most patients, ranging from 8 to 10 mm in length and from 2 to 5 mm in width. On MR imaging, the fibular avulsion was identified in 11 of the 13 patients. The other two patients had marrow edema in the fibular styloid process, although the avulsion fracture was not evident. All patients had injuries of the posterior cruciate ligament (six tibial avulsions, seven midsubstance tears). No patient had a tear of the anterior cruciate ligament. Disruption of the lateral collateral ligament was evident in seven patients, and one patient had a tear of the popliteal tendon. During surgery, six patients had disruption of the arcuate complex, but this disruption could not be identified on the MR images. CONCLUSION: An avulsion fracture of the fibular head generally involves the styloid process and causes injury of some of the major stabilizers in the posterolateral corner. Avulsion fractures are strongly associated with disruption of the posterior cruciate ligament. PMID- 12540439 TI - Sonography of the medial collateral ligament of the elbow: a study of cadavers and healthy adult male volunteers. AB - OBJECTIVE: We assessed the ability of high-resolution sonography to reveal the size and echogenicity of the anterior bundle of the normal medial collateral ligament of the elbow in cadavers and uninjured male volunteers. MATERIALS AND METHODS: The anterior bundle of the medial collateral ligament in five cadaveric elbows was imaged and injected with contrast material by experienced musculoskeletal radiologists using a 12-MHz linear array transducer. Immediate ligament dissection was performed. The bilateral ligaments in 30 healthy 21- to 34-year-old male volunteers were imaged with gravitational stress. Of these, five randomly selected subjects also received approximately 5 lb (11.3 kg) of applied stress. RESULTS: Contrast material was injected directly into all five cadaveric ligaments. The anterior bundle of the medial collateral ligament in all 30 asymptomatic male volunteers was hyperechoic in comparison with surrounding muscle and had a fibrillar pattern and fanlike shape. Its mean dimensions were 2.6 +/- 0.31 x 2.2 +/- 0.47 x 4.0 +/- 0.88 mm on the right and 2.6 +/- 0.36 x 2.1 +/- 0.42 x 4.0 +/- 0.86 mm on the left, in longitudinal short, transverse short, and transverse long axes, respectively. Differences in ligament measurements in sidedness, stress application, and hand dominance did not approach statistical (Bonferroni corrected, p > 0.01) or clinical (all differences, <0.2 mm) significance. CONCLUSION: Radiologists can accurately use sonography to identify and measure the size of the anterior bundle of the normal medial collateral ligament of the elbow. These baseline parameters for the normal ligament may prove useful when evaluating the injured ligament. PMID- 12540440 TI - Sonography of muscle hernias. PMID- 12540442 TI - Helical CT of the body: a survey of techniques used for pediatric patients. AB - OBJECTIVE: Our purpose was to assess the current practice of helical CT of the body in pediatric patients through a survey of members of the Society for Pediatric Radiology. MATERIALS AND METHODS: The survey consisted of 53 questions addressing demographics; oral and IV contrast media administration; and age-based (age groups, 0-4, 5-8, 9-12, and 13-16 years) scanning parameters, including tube current, kilovoltage, slice thickness, and pitch. Respondents accessed the Web based survey via a uniform resource locator link included in an e-mail to the members of the Society for Pediatric Radiology automatically sent every week for three weeks. Survey results were automatically tabulated. RESULTS: Most (83%) respondents were based in children's or university hospitals at the time of the survey. Virtually all (99%) used nonionic IV contrast material. For body scanning, 21-32% used less than 2.0 mL/kg of body weight; we found the percentage of respondents who used power injection to be approximately equal to the percentage of those who used manual injection (47%). Age-based adjustments are made; however, 11-26% of CT examinations of children younger than 9 years are performed using more than 150 mA. A notable finding was that 20-25% of respondents did not know specific parameters used for their examinations. CONCLUSION: Although pediatric radiologists do practice age-adjusted helical CT, variable scanning techniques are used, potentially delivering high doses of radiation. Information on current practices in helical CT of the body in children can serve as a foundation for future recommendations and investigations into helical CT in pediatric patients. PMID- 12540443 TI - In-plane bismuth breast shields for pediatric CT: effects on radiation dose and image quality using experimental and clinical data. AB - OBJECTIVE: The purpose of our study was to evaluate the amount of radiation dose reduction and its effect on image quality when using an in-plane bismuth breast shield for multidetector CT (MDCT) of the chest and abdomen in female pediatric patients. SUBJECTS AND METHODS: Fifty consecutive MDCT examinations (chest, 29; abdomen, 21) of female pediatric patients (mean age, 9 years; range, 2 months-18 years) were performed with a 2-ply (1.7 g of bismuth per square centimeter) bismuth shield (three sizes to accommodate patients of varying sizes) overlying the patient's breasts. MDCT images were evaluated for a perceptible difference in image quality in the lungs at the anatomic level under the shield as compared with nonshielded lung and whether the images were of diagnostic quality. In addition, 2-mm regions of interest were placed in the peripheral anterior and posterior portions of each lung in shielded and nonshielded areas, and noise (standard deviation in Hounsfield units) was measured in the regions. Differences among the regions in noise were compared for shielded versus nonshielded areas (paired t test). To measure differences in actual dose, we also evaluated the breast shield with an infant anthropomorphic phantom using thermoluminescent detectors in the breast tissue. The phantom was imaged with and without the breast shield using identical MDCT parameters. RESULTS: All MDCT scans of patients were of diagnostic quality with no perceptible difference in image quality in shielded versus nonshielded lung. We found no statistically significant difference in noise between the shielded and nonshielded lung regions of interest (shielded: mean noise, 17.3 H; nonshielded: mean noise, 18.8 H; p = 0.5180). Phantom measurements revealed a 29% reduction in radiation dose to the breast when a medium-dose MDCT protocol was used. CONCLUSION: Bismuth in-plane breast shielding for pediatric MDCT decreased radiation dose to the breast without qualitative or quantitative changes in image quality. PMID- 12540444 TI - MR imaging of spinal tumors in children with neurofibromatosis 1. AB - OBJECTIVE: The need for radiologic surveillance of spinal tumors in children with neurofibromatosis 1 is controversial and unresolved. We aimed to determine the incidence of spinal tumors in asymptomatic patients, describe the imaging findings, and evaluate the clinical usefulness of a prospective MR imaging surveillance program of the spine in children with neurofibromatosis 1. SUBJECTS AND METHODS: Of 62 children consecutively seen in a neurofibromatosis 1 clinic, 53 (85.5%) were recruited for MR imaging of the whole spine. All children satisfied the clinical criteria for diagnosis of neurofibromatosis 1. Imaging findings, clinical signs and symptoms, and subsequent clinical management were reviewed. RESULTS: The patients were 35 boys and 18 girls (age range, 11 months 18 years; mean age, 9.6 years), all of whom were asymptomatic, with no remarkable neurologic signs. Seven children (13.2%) had spinal neurofibromas: four had solitary neurofibromas (two dumbbell, one intradural, and one paraspinal tumor) and three had plexiform neurofibromas of the sacral plexus and thoracic and lumbar nerve sheaths. The incidences of scoliosis, localized cutaneous neurofibromas, and massive soft-tissue neurofibromas were 71.4%, 71.4%, and 28.6%, respectively, in the group with spinal neurofibromas (n = 7), and 30.4%, 39.1%, and 8.7%, respectively, in the group without spinal neurofibromas (n = 46). Patient clinical outcome was affected in only one patient (1.9%) in whom a solitary neurofibroma was resected. Follow-up imaging in 10 patients (mean period, 29 months) showed no evidence of tumor occurrence, progression, or recurrence. CONCLUSION: Although benign spinal neurofibromas are not uncommon in asymptomatic children with neurofibromatosis 1, the clinical usefulness of spine surveillance with MR imaging is limited in these children, making its effectiveness questionable. PMID- 12540445 TI - Interobserver reproducibility of volumetric MR imaging measurements of plexiform neurofibromas. AB - OBJECTIVE: This article validates the reproducibility of MR imaging volumetric measurements for evaluating the growth of plexiform neurofibromas in neurofibromatosis type 1. CONCLUSION: Volumetric measurements of plexiform neurofibromas in neurofibromatosis 1 are reproducible. PMID- 12540446 TI - Sonographic findings of Meckel's diverticulitis in children. AB - OBJECTIVE: We know of only one previous study that has described the sonographic appearances of the inflamed Meckel's diverticulum in several cases (nine patients). Our study reviews sonographic, clinical, and pathologic findings in 10 patients with an inflamed Meckel's diverticulum. We also describe unique findings on color Doppler sonography for two of our patients. CONCLUSION: Sonographic findings of an inflamed Meckel's diverticulum may mimic findings for acute appendicitis or intestinal duplication. In patients with rectal bleeding due to diverticulitis, the visualization of a tubular hyperechoic structure on sonography is suggestive of Meckel's diverticulum. The inflamed Meckel's diverticulum may present as a cyst, but its mucosal layer is more irregular than that found in an intestinal duplication. We found that routine color Doppler sonography revealed anomalous vessels and signs of inflammation on the wall of the Meckel's diverticulum. PMID- 12540447 TI - CT of renal lymphoma in children. AB - OBJECTIVE: The purpose of this study was to determine the incidence and to describe the CT findings of renal lymphoma in children. CONCLUSION: Renal lymphoma was seen in 11 children, most commonly due to non-Hodgkin's lymphoma (n = 10, 91%) and Burkitt's subtype (n = 5, 45%). Multiple bilateral masses were the most common CT appearance. Less common presentations included focal solitary masses or an engulfing mass. Retroperitoneal lymph node enlargement and other organ involvements were common associated findings. Five of six patients with renal involvement at initial diagnosis have had cures or remissions. Therefore, renal involvement at the time of initial diagnosis with lymphoma does not portend a poor prognosis. PMID- 12540448 TI - Prenatal sonography and MR imaging of pulmonary sequestration. PMID- 12540449 TI - MR imaging of hepatic pulmonary fusion in neonates. PMID- 12540450 TI - Imaging in the diagnosis, staging, treatment, and surveillance of hepatocellular carcinoma. PMID- 12540451 TI - Detection of hepatocellular carcinoma: comparison of dynamic MR imaging with dynamic double arterial phase helical CT. AB - OBJECTIVE: Three-dimensional (3D) Fourier transformation-enhanced fast gradient echo sequences with a special spectral inversion recovery pulse and fat suppression developed for abdominal imaging, including MR angiography, can show enhanced areas clearly. The purpose of this study was to evaluate the efficacy of dynamic MR imaging with the pulse sequences for the detection of hypervascular hepatocellular carcinoma by comparing it with that of dynamic helical CT with double arterial phase imaging. SUBJECTS AND METHODS: Fifty-three patients with 103 hypervascular hepatocellular carcinoma nodules who underwent both dynamic MR imaging with 3D Fourier transformation-enhanced fast gradient-echo sequences with a special spectral inversion recovery pulse and dynamic helical CT with double arterial phase imaging were enrolled in the study. For dynamic MR imaging, unenhanced, arterial, portal venous, and equilibrium phase images were obtained before and approximately 19, 60, and 120 sec, respectively, after injection of gadopentetate dimeglumine. Three observers independently interpreted the images obtained with each technique in a blinded manner and in random order. RESULTS: Mean sensitivity and positive predictive values of CT for hypervascular hepatocellular carcinoma (66% and 97%, respectively) were higher than those of MR imaging (63% and 96%, respectively), but there was no significant difference in detecting sensitivity among the observers (p < 0.05). CT and MR imaging were complementary, with some tumors undetected by CT but revealed on MR imaging. There was also no significant difference in A(z) values between CT (0.74) and MR imaging (0.71) (p < 0.05). CONCLUSION: Dynamic MR imaging with 3D Fourier transformation-enhanced fast gradient-echo sequences with a special spectral inversion recovery pulse is recommended to improve the detection of hypervascular hepatocellular carcinoma nodules in addition to the use of dynamic helical CT with double arterial phase imaging. PMID- 12540452 TI - Esophageal varices: diagnosis with gadolinium-enhanced MR imaging of the liver for patients with chronic liver damage. AB - OBJECTIVE: The purpose of our study was to evaluate the value of gadolinium enhanced MR imaging of the liver in the detection and grading of esophageal varices in patients with chronic liver damage. MATERIALS AND METHODS: Three independent off-site radiologists retrospectively reviewed MR images of the liver obtained in 72 patients with chronic liver damage, including 49 patients with untreated esophageal varices. All patients had undergone gastrointestinal endoscopy within 2 weeks of MR imaging. Both MR and endoscopic images were reviewed to determine whether esophageal varices were present and, if so, to determine the grade of the varices. Observer performances were tested with receiver operating characteristic curve analysis using the jackknife dispersion test. Correlations between the grades of the varices determined using MR images and those determined using endoscopic images were tested. RESULTS: Sensitivity for detection of esophageal varices was significantly (p < 0.01) higher for the combination of unenhanced and gadolinium-enhanced MR images (81%) than for the unenhanced MR images alone (51%). The receiver operating characteristic curve analysis (area under the curve, [A(z)]) showed that performance using the combination of the unenhanced and gadolinium-enhanced MR images (A(z) = 0.641) was superior to that using unenhanced MR images alone (A(z) = 0.586). A statistically significant positive correlation (p < 0.05) was found between the grades determined using MR imaging and the grades determined using endoscopy. CONCLUSION: Our results suggest the potential value of diagnosing the presence and grade of esophageal varices on MR imaging of the liver for patients with chronic liver damage. Gadolinium-enhanced MR imaging may increase the potential value. PMID- 12540453 TI - Pseudolesion of the bile duct caused by flow effect: a diagnostic pitfall of MR cholangiopancreatography. AB - OBJECTIVE: Our objective was to examine the influence of the shape of imaged structures and the velocity of flow on the appearance of flow artifacts seen on MR cholangiopancreatography (MRCP) in a phantom model. MATERIALS AND METHODS: Three types of phantoms representing the biliary system were constructed. The first phantom type was a straight tube; the second, a single tube in which the inlet and outlet diameters varied by a ratio of as much as 1:6; and the third, a tube that simulated a stricture in the biliary system and a gallstone. All experiments were repeated three times. RESULTS: We did not observe any flow artifacts in the experiments we performed with the straight tubes. A higher rate of flow resulted in decreased signal intensity in tubes simulating bile ducts; the decreased signal was most likely to be observed on images in which the speed of flow exceeded 5 mm/sec. Flow artifacts were seen only if the ratio between the inlet and outlet diameters was 1:4 or greater. Simulations of bile duct abnormalities--such as a 50% stricture or the presence of a gallstone--did not produce any flow artifacts. CONCLUSION: In our experiments, a flow artifact could be seen on images in which the ratio between the inlet and the outlet diameters in the phantom was equal to or greater than 1:4. This finding indicates that a flow artifact could be observed in dilated bile ducts on MRCP under clinical conditions. Knowing that a pseudo-filling defect can be caused by a flow artifact should help to prevent misinterpretation of MRCP images. PMID- 12540454 TI - Serial ethanol ablation of multiple hepatic cysts as an alternative to liver transplantation. PMID- 12540455 TI - CT and pathologic assessment of prospective nodal staging in patients with ductal adenocarcinoma of the head of the pancreas. AB - OBJECTIVE: The aim of our study was to compare the assessment of peripancreatic lymph nodes using CT with the gold standard of detailed histopathologic assessment of resected specimens in patients with pancreatic ductal adenocarcinoma. SUBJECTS AND METHODS: Sixty-two patients with presumed pancreatic carcinoma were prospectively studied with dual-phase contrast-enhanced helical CT, and images were interpreted in consensus by three radiologists. Complete surgical resection was performed in 28 patients. A detailed nodal classification system was used for radiologic, surgical, and pathologic staging in the nine patients whose final diagnosis at histology was pancreatic ductal adenocarcinoma. RESULTS: Forty lymph nodes were prospectively identified on CT in these nine patients. Two of 23 nodes (9%) measuring less than 5 mm in the short-axis diameter were malignant, four of 11 nodes (36%) measuring 5-10 mm were malignant, and one of six nodes (17%) larger than 10 mm was malignant. Using a short-axis diameter of greater than 10 mm as the criterion for nodal involvement, we found a sensitivity of 14% (1/7) and a specificity of 85% (28/33), with a positive predictive value of 17% (1/6), a negative predictive value of 82% (28/34), and an overall accuracy of 73% (29/40). Ovoid nodal shape, clustering of nodes, and the absence of a fatty hilum were not useful predictors of malignancy on CT. CONCLUSION: In resectable pancreatic ductal adenocarcinoma, CT is not accurate overall for the prediction of nodal involvement. In a patient with presumed pancreatic carcinoma that is considered to be resectable, the depiction on CT of peripancreatic nodes should not prevent attempted curative resection. PMID- 12540456 TI - Imaging of retained surgical sponges in the abdomen and pelvis. PMID- 12540457 TI - Barium impaction therapy for refractory colonic diverticular bleeding. PMID- 12540458 TI - MR features of abdominal splenosis. PMID- 12540459 TI - Is there an increased incidence of contralateral testicular cancer in patients with intratesticular microlithiasis? AB - OBJECTIVE: The objective of this study was to determine if there is an association between intratesticular microlithiasis and contralateral testicular cancer. MATERIALS AND METHODS: Retrospective review of a radiology database revealed 156 men who had undergone testicular sonography and orchiectomy for testicular cancer. Sonographic abnormalities were correlated with medical records and histopathology. Statistical significance was assessed using Fisher's exact test. RESULTS: Twenty-three (15%) of 156 patients with prior orchiectomy for testicular cancer had microlithiasis, and 133 (85%) did not have microlithiasis. Four of 23 patients with microlithiasis had masses, and eight had heterogeneous changes. Sonograms of 133 patients without microlithiasis revealed masses in seven and heterogeneous changes in 15 patients. Five patients with microlithiasis and six without microlithiasis underwent a second orchiectomy. Contralateral testicular cancer was confirmed in five (22%) of 23 patients with microlithiasis versus three (2%) of 133 men without microlithiasis. Microlithiasis was present in five (63%) of eight patients with bilateral testicular cancer, and microlithiasis was highly associated with confirmed bilateral testicular cancer (5/23 vs 3/133, odds ratio [OR] = 12.0, p = 0.002). Among the 34 patients who had either testicular masses or heterogeneous changes, microlithiasis had an OR of 4.5 (p = 0.10). CONCLUSION: In our study, contralateral testicular cancer was significantly associated with intratesticular microlithiasis. Nevertheless, there was not sufficient evidence that intratesticular microlithiasis adds independent diagnostic information for bilateral testicular cancer in the absence of a mass or heterogeneous changes. PMID- 12540460 TI - Testicular Leydig's cell hyperplasia: MR imaging and sonographic findings. PMID- 12540461 TI - Trauma cases from Harborview Medical Center. Rupture of a ureteropelvic junction obstructed kidney in a 15-year-old football player. PMID- 12540462 TI - Three-dimensional coronary MR angiography performed with subject-specific cardiac acquisition windows and motion-adapted respiratory gating. AB - OBJECTIVE: In coronary MR angiography, data are conventionally accepted in only short and fixed periods of the cardiac and respiratory cycles. We hypothesized that a more flexible and subject-specific approach to cardiac and respiratory gating may shorten scanning times while maintaining image quality. SUBJECTS AND METHODS: We implemented an acquisition technique that uses subject-specific acquisition windows in the cardiac cycle and a motion-adapted gating window for respiratory navigator gating. Cardiac acquisition windows and trigger delays were determined individually from a coronary motion scan. Motion-adapted gating used a 2-mm acceptance window for the central 35% of k-space and a 6-mm window for the outer 65% of k-space. In 10 subjects, three-dimensional coronary MR angiograms of the right and left coronary systems were acquired with this technique (the "adaptive technique") as well as a conventional acquisition method, and the scanning times and image quality were compared. The adaptive technique was then applied prospectively to 40 patients who underwent coronary radiographic angiography. RESULTS: Scanning times with the adaptive technique were reduced by a factor of 2.3 for the right coronary artery and by a factor of 2.2 for the left coronary artery system compared with the conventional technique, mainly because we were able to use longer subject-specific acquisition windows in patients with low heart rates. Subjective and objective measurements of image quality showed no significant differences between the two techniques. Prospective evaluation of MR angiograms yielded a sensitivity and specificity of 74.3% and 88.2%, respectively, to detect significant coronary artery stenoses. CONCLUSION: Coronary MR angiography with subject-specific acquisition windows and motion adapted respiratory gating reduces scanning times while maintaining image quality and provides high diagnostic accuracy for the detection of coronary artery stenosis. PMID- 12540463 TI - Bronchoarterial ratio and bronchial wall thickness on high-resolution CT in asymptomatic subjects: correlation with age and smoking. AB - OBJECTIVE: The purpose of this study was to determine whether the values of the bronchoarterial ratio and bronchial wall thickness, as viewed during high resolution CT, relate to age and smoking status in asymptomatic healthy subjects. SUBJECTS AND METHODS: High-resolution CT was performed prospectively in 85 subjects without cardiopulmonary disease. The subjects were divided into three groups according to age: 29 subjects were 21-40 years old; 29 subjects, 41-64 years old; and 27 subjects, 65 years or older. Both bronchoarterial ratios, defined as the diameter of the bronchial lumen divided by the diameter of its accompanying artery, and the T/D ratio, defined as wall thickness (T) divided by the total diameter of the bronchus (D), were measured at the segmental and subsegmental levels of the apical and posterior basal segments. Each calculated ratio was evaluated according to age and smoking status. RESULTS: Significant correlation was found between the bronchoarterial ratio and age (r = 0.768, p < 0.0001), with the bronchoarterial ratio increasing with age and exceeding 1 in 41% of subjects older than 65 years. No significant correlation was seen between the T/D ratio and age. No significant differences in bronchoarterial ratio and T/D ratio were seen between smokers and nonsmokers in subjects overall; but in the elderly group, the T/D ratio was significantly higher in smokers than in nonsmokers (p = 0.021). CONCLUSION: The bronchoarterial ratio is influenced by aging. The normal bronchoarterial ratio in a substantial number of subjects older than 65 years overlaps with the ratio considered to represent bronchiectasis. Thus, when this ratio is used for the quantitative analysis of pulmonary and cardiovascular disease, the influence of age should be considered. PMID- 12540464 TI - Thymic enlargement and FDG uptake in three patients: CT and FDG positron emission tomography correlated with pathology. AB - OBJECTIVE: Our purpose was to describe three adult patients in whom we found increased thymic uptake of FDG on positron emission tomography and thymic enlargement with convex lateral margins on CT. Subsequent biopsy or resection showed normal thymic tissue. CONCLUSION: In three adults, we found a physiologic uptake of FDG by the thymus with standardized uptake values in the range of thymic neoplasia. PMID- 12540465 TI - Nonbronchial collateral supply from the hepatic arteries of a patient with hemoptysis. PMID- 12540466 TI - Experience at a single institution with endovascular treatment of mechanical complications caused by implanted central venous access devices in pediatric and adult patients. AB - OBJECTIVE: Our objective was to describe the technical aspects and evaluate the feasibility, safety, and efficacy of endovascular management of mechanical complications related to implanted central venous devices. MATERIALS AND METHODS: One hundred fifty-six patients with cancer, who ranged in age from 3 months to 75 years (mean +/- SD, 47 +/- 18 years), were referred 290 +/- 200 days (mean +/- SD; range, 0-1202 days) after central venous device placement for retrieval of a fractured and embolized central venous device catheter (n = 100), retrieval of a guidewire embolized during placement of a central venous device (n = 2), repositioning of the migrated tip of a central venous device catheter (n = 38), and fibrin-sheath stripping (n = 16). All procedures were performed with the patient under local anesthesia on an outpatient basis, except for the eight pediatric patients. RESULTS: Ninety-five of the 100 embolized catheters and both of the guidewires were successfully retrieved. Retrieval was preceded by repositioning the embolized catheter with a pigtail catheter in 48 of these cases. Most of the procedures were performed with standard vascular tools (loop snares and pigtail catheters); the use of more sophisticated devices (grasping forceps, baskets, or balloons) rarely overcame the failure of a loop snare. Repositioning a migrated catheter tip was achieved with a pigtail catheter in 32 of 38 attempts. Of the repositioned catheters, only 24 could be used. Most of the 11 repositioning and retrieval failures were encountered because the catheter lacked a free end. Fibrin-sheath stripping was always technically successful: all these catheters were patent at 3-month follow-up. No procedure-related complications occurred. CONCLUSION: The endovascular approach is highly feasible, safe, and effective for the management of mechanical complications of central venous devices. It is probably advisable to reserve endovascular repositioning for port catheters that are cumbersome to exchange and to replace simple catheters. PMID- 12540467 TI - Feasibility study of NeoMend, a percutaneous arterial closure device that uses a nonthrombogenic bioadhesive. AB - OBJECTIVE: The aim of this prospective single-center phase I feasibility study was to investigate the safety and efficacy of a novel vascular sealing device, the NeoMend Arterial Closure Device, that uses a bioadhesive after percutaneous endovascular procedures. SUBJECTS AND METHODS: In 26 consecutive patients, the sealing device was deployed at the femoral artery access site immediately after a catheterization procedure using a 6-French (1.91-mm) sheath. Patients were followed up at 24 hr with Doppler sonography of the treated femoral artery puncture site, and at 1 week and 1 month by a telephone interview. RESULTS: Successful hemostasis was achieved with the NeoMend Arterial Closure Device in 21 (88%) of 24 patients. One major complication required surgery: formation of puncture site hematoma and pseudoaneurysm 3 days after the intervention after successful primary hemostasis. Two device failures required crossover to manual compression, which was done without further complications. The mean time to hemostasis was 7.0 +/- 4.5 min. Mean time to ambulation was 6.0 hr. At follow-up, the patients did not report any puncture-site-related complaints. Doppler sonography of the puncture sites revealed three insignificant hematomas of less than 20 mL and patent common femoral vessels without stenoses. CONCLUSION: The NeoMend Arterial Closure Device appears to achieve rapid hemostasis with the potential of early ambulation after arterial punctures with a 6-French sheath. The device is an alternative in situations in which suture- or collagen-mediated devices show high complication rates. PMID- 12540468 TI - Is it a new pattern of pulmonary aspergillosis? PMID- 12540469 TI - Radiation risk of screening CT. PMID- 12540470 TI - Communication of the ganglion with the joint and the tendon sheath. PMID- 12540471 TI - Fluid removal limitations in thoracentesis. PMID- 12540473 TI - Acute bronchospasm due to exposure to polymethylmethacrylate vapors during percutaneous vertebroplasty. PMID- 12540474 TI - Detection on CT angiography of an accessory middle cerebral artery simulating a fusiform aneurysm on MR angiography. PMID- 12540475 TI - Dense coronary sinus: a sign of left superior vena cava on abdominopelvic CT. PMID- 12540476 TI - Multifocal involvement of the pancreas in multiple myeloma: sonographic, CT, and MR imaging findings. PMID- 12540477 TI - Epithelial cell modulation of airway fibrosis in asthma. PMID- 12540478 TI - Building a better heparin. PMID- 12540479 TI - Lung vascular development: breathing new life into an old problem. PMID- 12540480 TI - Airway neural plasticity: the nerves they are a-changin'. PMID- 12540481 TI - Mechanical stress triggers selective release of fibrotic mediators from bronchial epithelium. AB - Transforming growth factor-beta (TGF-beta) and endothelin (ET) are found in elevated amounts in the airways of individuals with asthma. The cellular source of these peptides and their role in mediating the airway fibrosis of chronic asthma are unknown. In response to mechanical stresses similar to those occurring in vivo during airway constriction, bronchial epithelial cells increase the steady-state level of mRNA for both ET-1 and ET-2, followed by increased release of ET protein. Mechanical stress also enhances release of TGF-beta2 from a preformed cell-associated pool. TGF-beta2 and ET act individually and, more importantly, synergistically to promote fibrotic protein synthesis in reporter fibroblasts. To confirm the role of these intermediates in stress-induced fibrosis, conditioned medium from mechanically stressed bronchial epithelial cells was shown to elicit fibrotic protein synthesis in reporter fibroblasts; this effect was significantly inhibited by combined treatment with ET receptor antagonists and a neutralizing antibody to TGF-beta2. These data are consistent with a primary pathogenic role for mechanical stress-induced release of both TGF beta2 and ET in the subepithelial fibrosis that characterizes chronic asthma. PMID- 12540482 TI - Effect of covalent serpin-heparinoid complexes on plasma thrombin generation on fetal distal lung epithelium. AB - Extravascular coagulation within the lung airspace is a hallmark of respiratory distress syndrome (RDS) in premature infants. We previously showed that covalent antithrombin-heparin complex (ATH) is superior to noncovalent antithrombin (AT) + heparin (H) mixtures at inhibiting plasma thrombin generation on rat fetal distal lung epithelium (FDLE) in vitro. However, heparin cofactor II (HC) has been shown to selectively inhibit thrombin, which may be advantageous if other enzyme activities are present in the airspace. We compared the abilities of ATH, covalent HC-heparin complex (HCH), and covalent HC-dermatan sulfate (HCD) to inhibit thrombin generation on FDLE in plasmas from either adults or newborns. In the presence of ATH, peak free thrombin generation in adult plasma on the cell surface was reduced by 92% compared with controls (no anticoagulant). However, whereas HCH reduced peak free thrombin generation in adult plasma by 81%, HCD was only able to reduce activity by 33%. All covalent complexes caused a greater decrease in thrombin activity compared with that with the corresponding noncovalent serpin + heparinoid mixtures. Experiments in plasma from newborns resulted in peak free thrombin that was less than or equal to that in adult plasma when covalent conjugates were added. Relative peak free thrombin was proportional to rate of prothrombin consumption and amount of thrombin-inhibitor complexes formed. In vivo, experiments in newborn rats showed that a greater percentage of intratracheally instilled ATH and HCH could be recovered in lung lavage fluid compared withwith that for HCD. In summary, ATH, HCH, and HCD are inhibitors of thrombin generation on FDLE superior to the corresponding noncovalent mixtures, with ATH and HCH being more potent than HCD. Covalent conjugates of AT or HC with H may be preferred in treatment of extravascular coagulation. PMID- 12540483 TI - Insulin-like growth factor-I receptor-mediated vasculogenesis/angiogenesis in human lung development. AB - The structural and functional development of the pulmonary system is dependent upon appropriate early vascularization of the embryonic lung. Our previous in vitro studies in a rat model indicated that insulin-like growth factor-I (IGF-I) is a potent angiogenic agent for fetal lung endothelial cells. To assess its role on human vascular lung development, we first examined the expression of IGF-I/II and IGF receptor type I (IGF-IR) in human embryonic and fetal lung tissues at 4 12 wk of gestation. Immunohistochemical and in situ hybridization studies revealed the presence of IGF-I/II-IGF-IR ligands and mRNA transcripts in embryonic lungs as early as 4 wk gestation. Immunotargeting using an anti-IGF-IR neutralizing antibody on human fetal lung explants demonstrated a significant blockade of IGF-IR signaling. Inactivation of IGF-IR resulted in a loss of endothelial cells, accompanied by dramatic changes in fetal lung explant morphology. Terminal transferase dUTP end-labeling assay and TEM studies of anti IGF-IR-treated lungs demonstrated numerous apoptotic mesenchymal cells. Rat embryonic lung explant studies further validated the importance of the IGF-IGF-IR system for lung vascular development. These data provide the first demonstration of IGF-I/II expression in the human lung in early gestation and indicate that the IGF family of growth factors, acting through the IGF-IR, is required as a survival factor during normal human lung vascularization. PMID- 12540484 TI - Pan-neurotrophin receptor p75 contributes to neuronal hyperreactivity and airway inflammation in a murine model of experimental asthma. AB - Bronchial asthma represents a severe chronic inflammatory disease with increasing prevalence. The pathogenesis is characterized by complex neuroimmune dysregulation. Although the immunopathogenesis of the disease has been extensively studied, the nature of neuronal dysfunction still remains poorly understood. Recent data indicate that neurotrophins contribute to airway inflammation, broncho-obstruction and airway hyperresponsiveness. Using an established murine model of allergic bronchial asthma, the contribution of the pan-neurotrophin receptor p75(NTR) was defined. This receptor is expressed both in normal and asthmatic lungs and airways. Analysis of p75(NTR-/-) mice, as well as in vivo blocking of p75(NTR), revealed that airway inflammation is to a large extent dependent upon functional receptor expression. Furthermore, neuronal hyperreactivity depends entirely on this receptor. Based on these data, a novel molecular pathway in the neuroimmune pathogenesis of bronchial asthma could be defined. PMID- 12540485 TI - Hypoxia protects human lung microvascular endothelial and epithelial-like cells against oxygen toxicity: role of phosphatidylinositol 3-kinase. AB - Hypoxic preconditioning is protective against oxidant-related damage in various organs, such as the heart. We previously showed that rats exposed to hypoxia also exhibit resistance to lethal pulmonary oxygen toxicity. The underlying mechanism and whether similar preconditioning is applicable to cellular models is unknown. In the present study, it was found that hypoxic pre-exposure induces a significant protective effect against hyperoxia-induced cell death in human lung microvascular endothelial cells (HLMVECs) and epithelial type II-like A549 cells. This effect of hypoxia is mediated by the phosphatidylinositol 3-kinase (PI3-K) signaling pathway because the presence of the PI3-K inhibitors, LY294002 and wortmannin, during pre-exposure to hypoxia completely blocks subsequent protection. Further, the hypoxia-dependent protection from hyperoxia was found to be associated with a 2-fold increase in PI3-K activity in hypoxia. Transient overexpression of a catalytically active class IA PI3-K p110alpha isoform also enhanced survival of A549 cells 2-fold compared with the empty vector control. These results indicate that hypoxia-induced activation of PI3-K is an important event in the acquisition of resistance against subsequent hyperoxic toxicity. PMID- 12540486 TI - Inhibition of nitric oxide restores surfactant gene expression following nickel induced acute lung injury. AB - The role of nitric oxide (NO) in acute lung injury remains controversial. Although inhaled NO increases oxygenation in clinical trials, inhibiting NO synthase (NOS) can be protective. To examine the latter, nickel-exposed mice were treated with saline or NOS inhibitor, N(G)-nitro-L-arginine methyl ester (L NAME). Initial microarray analysis of nickel-induced gene expression of saline treated mice revealed increased inflammatory mediator, matrix injury-repair, and hypoxia-induced factor-mediated sequences and decreased lung-specific (e.g., surfactant-associated protein B and C) sequences. Compared with saline control, L NAME-treated mice had enhanced survival with attenuated serum nitrate/nitrite, endothelial NOS activity, and lavage neutrophils and protein. Although initial cytokine (i.e., interferon-gamma, interleukins-1beta and -6, macrophage inflammatory protein-2, monocyte chemotactic protein-1, and tumor necrosis factor alpha) gene expression was similar between groups, subsequent larger cytokine increases only occurred in saline-treated mice. Similarly, surfactant protein gene expression decreased initially in both groups yet was restored subsequently with L-NAME treatment. Interestingly, the role of inducible NOS (iNOS) in these responses seems minimal. iNOS gene expression was unaltered, iNOS activity and nitrotyrosine residues were undetectable, and an iNOS antagonist, aminoguanidine, failed to increase survival. Rather, systemic L-NAME treatment appears to attenuate pulmonary endothelial NOS activity, subsequent cytokine expression, inflammation, and protein permeability, and thereby restores surfactant gene expression and increases survival. PMID- 12540487 TI - Vitamin E reduces transendothelial migration of neutrophils and prevents lung injury in endotoxin-induced airway inflammation. AB - We investigated the pharmacologic effects of the antioxidant Vitamin E (alpha tocopherol [alpha-toc]) in airway inflammation induced by inhaled endotoxin. A preparation of alpha-toc incorporated in liposomes was administered intraperitoneally in mice 1 h after exposure of aerosolized endotoxin. Injection of 50 mg alpha-toc/kg significantly decreased the number of neutrophils in airspaces and prevented lung injury, monitored both as decreased lactate dehydrogenase activity in airways and reduced lung edema when compared with animals treated with plain liposomes. Immunofluorescence staining of lung tissue revealed that treatment with alpha-toc decreased the number of neutrophils in lung interstitium, whereas the number in lung blood vessels and peripheral blood did not differ between mice treated with alpha-toc and control mice. Our results indicate that alpha-toc downmodulates the migration of neutrophils across the endothelial barrier, but in contrast to strong anti-inflammatory drugs such as corticosteroids, without inhibition of transcription factors involved in the early inflammatory response (nuclear factor-kappaB/activator protein-1). Neither was the endotoxin-induced expression of proinflammatory cytokines in lung tissue downregulated. Treatment with a combination of alpha-toc and a suboptimal dose of 0.5 mg/kg dexamethasone enhanced the effect, suggesting that alpha-toc, in combination with low doses of corticosteroids, might be effective for therapeutic treatment of acute lung injury. PMID- 12540488 TI - Differential role for T cells in the development of fibrotic lesions associated with reovirus 1/L-induced bronchiolitis obliterans organizing pneumonia versus Acute Respiratory Distress Syndrome. AB - Bronchiolitis obliterans organizing pneumonia (BOOP) and Acute Respiratory Distress Syndrome (ARDS) are two pulmonary diseases with fibrotic components. BOOP is characterized by perivascular/peribronchiolar leukocyte infiltration leading to the development of intra-alveolar fibrosis. ARDS is a biphasic disease that includes an acute phase, consisting of severe leukocyte infiltration, edema, hemorrhage, and the formation of hyaline membranes, and a chronic phase, which is characterized by persistent intra-alveolar and interstitial fibrosis. CBA/J mice infected with 1 x 10(6) plaque-forming units (pfu) reovirus 1/L develop follicular bronchiolitis and intra-alveolar fibrosis similar to BOOP. In contrast, CBA/J mice infected with 1 x 10(7) pfu reovirus 1/L develop histologic characteristics of ARDS including diffuse alveolar damage, hyaline membranes, and intra-alveolar fibrosis. In this report, we demonstrate a differential role for T lymphocytes in the development of fibrosis associated with BOOP versus ARDS. Neonatally thymectomized CBA/J mice infected with 1 x 10(7) pfu (ARDS) reovirus 1/L still develop the hallmark characteristics of ARDS, including a severe viral pneumonia with cellular infiltrates comprised mainly of macrophages and neutrophils, hyaline membrane formation, and hemorrhage during the acute phase of the disease and persistent intra-alveolar fibrosis during the chronic phase of the disease. In contrast, neonatally thymectomized CBA/J mice infected with 1 x 10(6) pfu (BOOP) reovirus 1/L do not develop intra-alveolar fibrosis associated with BOOP. Therefore, while T cells are necessary for the development of intraluminal fibrosis associated with BOOP, they are not necessary for the development of intraluminal fibrosis associated with ARDS. Furthermore, we suggest that interferon-gamma plays a key role in the fibrotic process and that elevated levels of interferon-gamma are associated with a continuum from least to more severe fibrosis. PMID- 12540489 TI - High tidal volume ventilation induces proinflammatory signaling in rat lung endothelium. AB - Alveolar overdistension during mechanical ventilation causes leukocyte sequestration, leading to lung injury. However, underlying endothelial cell (EC) mechanisms are undefined. In a new approach, we exposed isolated blood-perfused rat lungs to high tidal volume ventilation (HV) for 2 h, then obtained fresh lung endothelial cells (FLEC) by immunosorting at 4 degrees C. Immunoblotting experiments indicated that as compared with FLEC derived from lungs ventilated at low volume (LV), HV markedly enhanced tyrosine phosphorylation (TyrP). The tyrosine kinase blocker, genistein, inhibited this response. HV also induced focal adhesion (FA) formation in FLEC, as detected by immunofluorescent aggregates of the alpha(v)beta(3) integrin that co-localized with aggregations of focal adhesion kinase (FAK). Immunoprecipitation and blotting experiments revealed that HV increased TyrP of the FA protein, paxillin. In addition, HV induced a paxillin-associated P-selectin expression on FLEC that was also inhibited by genistein. However, HV did not increase lung water. These results indicate that in HV, EC signaling in situ causes FA formation and induces TyrP dependent P-selectin expression. These signaling mechanisms may promote leukocyte mediated responses in HV. PMID- 12540490 TI - Tumor necrosis factor-related apoptosis-inducing ligand enhances collagen production by human lung fibroblasts. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/APO-2L) is a member of the tumor necrosis factor family that induces apoptosis in a variety of transformed cell lines and in normal human hepatocytes and brain cells. Soluble TRAIL at high concentrations was found to induce apoptotic death in normal human lung fibroblasts, whereas at low concentrations it was found to stimulate collagen production by these cells. Collagen alpha2(I) mRNA expression was assessed by semiquantitative reverse transcriptase/polymerase chain reaction; total soluble collagen was measured in culture supernatants by the Sircol assay. Both alpha2(I) collagen mRNA level and total soluble collagen secretion were increased upon TRAIL stimulation, with peak response (> 4-fold increase in mRNA level) at 1 ng/ml TRAIL. Analysis of the transcriptional response in TRAIL stimulated fibroblasts, using DNA microarray hybridization, revealed an augmented expression of a number of genes involved in tissue remodeling, including those related to the transforming growth factor-beta (TGF-beta) pathway. DNA microarray results for the increase in TGF-beta1 mRNA level were confirmed by Northern blot analysis and by measurements of total active TGF-beta1 in culture supernatants. In addition, pan-specific TGF-beta antibody was shown to inhibit TRAIL-stimulated collagen mRNA and protein expression. These data suggest that TRAIL can enhance extracellular matrix synthesis in fibroblasts by triggering TGF-beta production that acts in an autocrine manner. PMID- 12540491 TI - Antisense oligodeoxynucleotides decrease LGL1 mRNA and protein levels and inhibit branching morphogenesis in fetal rat lung. AB - We previously described the cloning of the late gestation lung 1 gene (LGL1), a novel glucocorticoid-inducible gene expressed in the mesenchyme of fetal lung. We report here evidence for a role of the LGL1 gene product (lgl1) in fetal rat lung airway branching morphogenesis, temporal and spatial localization of LGL1 mRNA and lgl1 protein in fetal rat lung, and a correction of the previously published LGL1 sequence. Both the mRNA and protein were detected during fetal lung development. LGL1 mRNA was detected from gestational Day 12 by reverse transcriptase-polymerase chain reaction, and from Day 13 by in situ hybridization. lgl1 protein was detected from Day 18 by Western analysis and from Day 16 by immunohistochemistry. The types of cells expressing LGL1 mRNA and lgl1 protein were assessed by immunohistochemical staining of adjacent serial tissue sections for markers of mesenchymal (vimentin) and smooth muscle (alpha-actin) cells. As gestation advanced, increasing amounts of mRNA and protein were expressed in these cells. In support of a role for lgl1 in airway branching morphogenesis, antisense (but neither sense nor scrambled) oligodeoxynucleotides directed against LGL1 inhibited airway branching in fetal rat lung buds in explant culture, in a dose- and time-dependent manner. The levels of lgl1 protein and LGL1 mRNA expression were decreased in those explants that had inhibited airway branching, compared with the uninhibited controls. Our findings suggest that lgl1 plays an important role in fetal airway branching morphogenesis. PMID- 12540492 TI - The mitochondria-regulated death pathway mediates asbestos-induced alveolar epithelial cell apoptosis. AB - The mechanisms underlying asbestos-induced pulmonary toxicity are not fully understood. Alveolar epithelial cell (AEC) apoptosis by iron-derived reactive oxygen species (ROS) is one important mechanism implicated. The two major pathways regulating apoptosis include (i) the mitochondrial death (intrinsic) pathway caused by DNA damage, and (ii) the plasma-membrane death receptor (extrinsic) pathway. However, it is unknown whether asbestos activates either death pathway in AEC. We determined whether asbestos triggers AEC mitochondrial dysfunction by exposing cells (A549 and rat alveolar type II) to amosite asbestos and assessing mitochondrial membrane potential changes (deltapsi(m)) using a fluorometric technique involving tetremethylrhodamine ethyl ester (TMRE) and mitotracker green. Unlike inert particulates (titanium dioxide and glass beads), amosite asbestos caused dose- and time-dependent reductions in deltapsi(m). Asbestos-induced deltapsi(m) was associated with the release of cytochrome c from the mitochondria to the cytoplasm as well as activation of caspase 9, a mitochondrial-activated caspase. In contrast, a lower level of caspase 8, the death receptor-activated caspase, was detected in asbestos-exposed AEC. An iron chelator (phytic acid or deferoxamine) or a hydroxyl radical scavenger (sodium benzoate) each blocked asbestos-induced reductions in deltapsi(m) and caspase 9 activation, suggesting a role for iron-derived ROS. Finally, Bcl-X(L), a mitochondrial antiapoptotic protein that prevents cell death by preserving the outer mitochondrial membrane integrity, blocked asbestos-induced decreases in A549 cell deltapsi(m) and reduced apoptosis as assessed by DNA fragmentation. We conclude that asbestos-induced AEC apoptosis results from mitochondrial dysfunction, in part due to iron-derived ROS, which is followed by the release of cytochrome c and caspase 9 activation. Our findings suggest an important role for the mitochondria-regulated death pathway in the pathogenesis of asbestos associated pulmonary toxicity. PMID- 12540493 TI - Surfactant protein A and D differently regulate the immune response to nonmucoid Pseudomonas aeruginosa and its lipopolysaccharide. AB - We investigated the role of the surfactant proteins (SPs) A and D in the pulmonary immune defense of nonmucoid strains of Pseudomonas aeruginosa, the most etiologic agents of nosocomial Pseudomonas pneumonia. We first examined the interactions of recombinant human SP-D dodecamers and purified natural or recombinant human SP-A with two smooth, and two rough, clinical isolates of nonmucoid P. aeruginosa. SP-D bound to all four isolates, but agglutinated only one rough and one smooth strain. SP-D functioned as an opsonin to enhance the uptake of all four strains by the human monocytic cell line Mono Mac 6 (MM6). SP D also enhanced tumor necrosis factor-alpha secretion by MM6 cells in response to purified lipopolysaccharide (LPS) isolated from the rough, but not the smooth, strains. Although SP-A bound to all four strains, it did not cause bacterial aggregation or enhance uptake. It showed small but statistically significant inhibitory effects on the cytokine response of MM6 cells to one strain of smooth organisms, but did not significantly alter the response to purified LPS. This study in combination with previously published data strongly suggests that SP-D may play important roles in the local innate pulmonary defense against nonmucoid P. aeruginosa of diverse LPS phenotypes, and preferentially augments the cellular response to rough P. aeruginosa endotoxin. PMID- 12540494 TI - Muscarinic M3-receptors mediate cholinergic synergism of mitogenesis in airway smooth muscle. AB - Muscarinic receptor agonists have been considered to act synergistically in combination with growth facors on airway smooth muscle growth. Characterization of the proliferative responses and of the receptor subtype(s) involved has not yet been studied. Therefore, we investigated mitogenesis induced by stimulation of muscarinic receptors, alone and in combination with stimulation by platelet derived growth factor (PDGF). For this purpose, [(3)H]thymidine-incorporation was measured at different culture stages in bovine tracheal smooth muscle cells. Functional muscarinic M(3)-receptors, as measured by formation of inositol phosphates, were present in unpassaged cells, but were lacking in passage 2 cells. Methacholine (10 microM) by itself was not able to induce a proliferative response in both cell culture stages. However, methacholine interacted synergistically with PDGF in a dose-dependent fashion (0.1-10 microM), but only in cells having functional muscarinic M(3)-receptors. This synergism could be suppressed significantly by the selective M(3)-receptor antagonists DAU 5884 (0.1 microM) and 4-DAMP (10 nM), but not at all by the M(2)-subtype selective antagonist gallamine (10 microM). These results show that methacholine potentiates mitogenesis induced by PDGF solely through stimulation of muscarinic M(3)-receptors in bovine tracheal smooth muscle cells. PMID- 12540496 TI - Global control of primary hepatocellular carcinoma with hepatitis B vaccine: the contributions of research in Taiwan. PMID- 12540497 TI - A framework for improving the quality of cancer care: the case of breast and cervical cancer screening. AB - This commentary presents a conceptual framework, Quality in the Continuum of Cancer Care (QCCC), for quality improvement studies and research. Data sources include review of relevant literature (cancer care, quality improvement, organizational behavior, health services evaluation, and research). The Detecting Early Tumors Enables Cancer Therapy (DETECT) project is used to apply the QCCC model to evaluate the quality of secondary prevention. Cancer care includes risk assessment, primary prevention, screening, detection, diagnosis, treatment, recurrence surveillance, and end-of-life care. The QCCC model represents a systematic approach for assessing factors that influence types of cancer care and the transitions between them, the factors at several levels (community, plan and practice setting) that potentially impact access and quality, and the strategies groups and organizations can consider to reduce potential failures. Focusing on the steps and transitions in care where failures can occur can facilitate more organized systems and medical practices that improve care, establish meaningful measures of quality that promote improved outcomes, and enhance interdisciplinary research. PMID- 12540498 TI - Genetic polymorphisms in GSTM1, -P1, -T1, and CYP2E1 and the risk of adult brain tumors. AB - GST and CYP2E1 genes are involved in metabolism of several compounds (e.g., solvents) that may play a role in brain cancer etiology. We evaluated associations between polymorphisms in these genes and adult brain tumor incidence. Cases were 782 patients with brain tumors diagnosed from 1994 to 1998 at three United States hospitals. Controls were 799 patients admitted to the same hospitals for nonmalignant conditions. DNA was extracted from blood samples that had been collected from 1277 subjects (80% of all subjects; 604 controls; 422 gliomas, 172 meningiomas, and 79 acoustic neuromas), and genotyping was successfully conducted for GSTM1 null, GSTT1 null, GSTP I105V, GSTP A114V, CYP2E1 RsaI, and CYP2E1 Ins96. The GSTP1 105 Val/Val genotype was associated with increased glioma incidence [odds ratio (OR), 1.8; 95% confidence limits (CLs), 1.2, 2.7], with the estimated effect following a trend of increasing magnitude by number of variant alleles (Ile/Ile: OR, 1.0; Ile/Val: OR, 1.3; Val/Val: OR, 2.1). The CYP2E1 RsaI variant was weakly associated with glioma (OR, 1.4; 95% CL, 0.9, 2.4) and acoustic neuroma (OR, 2.3; 95% CL, 1.0, 5.3), with some indication of stronger associations among younger subjects. Estimated effects of the gene variants differed by glioma subtype. There was evidence of supermultiplicativity of the joint effect of GSTP1 I105V and CYP2E1 RsaI variants on both glioma and acoustic neuroma, even following adjustment of estimates toward a common prior distribution using hierarchical regression models. Previously reported associations between the GSTT1 null genotype and overall glioma incidence were not replicated, but an association with meningioma was observed (OR, 1.5; 95% CL, 1.0, 2.3). These findings may provide clues to both genetic and environmental determinants of brain tumors. PMID- 12540499 TI - Vitamin D receptor start codon polymorphism (FokI) and prostate cancer progression. AB - Vitamin D plays an important role in cell growth and differentiation and is proposed to protect against cancer initiation and/or progression. The vitamin D receptor (VDR) has a thymine/cytosine (T/C) polymorphism located in the first of two potential start (ATG) codons that can be detected by a RFLP using the endonuclease FokI. The C variant, which lacks the first ATG, results in a shorter VDR and is referred to as the F allele. The T variant (f allele) initiates at the first ATG. We examined the association of the VDR FokI genotype with histopathological characteristics and prognosis of prostate cancer among 191 mostly Caucasian subjects who had undergone radical prostatectomy between 1984 and 1992. The frequencies of the FF, Ff, and ff genotypes were 41%, 38%, and 21%, respectively. Subjects with the ff genotype had a lower mean percentage of Gleason grade 4/5 cancer (30.3%) than subjects with the FF or Ff genotypes (42.8% and 43.8%, respectively; P = 0.015 by t test for ff versus FF + Ff). The data suggest that the presence of an F allele increased the risk of being diagnosed with more aggressive cancer because higher percentage of Gleason grade 4/5 is associated with worse prognosis. The age-adjusted risk of prostate-specific antigen failure was lower for the ff genotype than for the FF genotype by Cox proportional hazards analysis but did not achieve statistical significance (hazard ratio = 0.76; 95% confidence interval, 0.44-1.32). This risk reduction disappeared after further adjustment for percentage of Gleason grade 4/5, cancer volume, and preoperative serum prostate-specific antigen level (hazard ratio = 1.03; 95% confidence interval, 0.58-1.85). In conclusion, the ff genotype was associated with less aggressive histopathological findings than Ff or FF genotypes. Additional studies with a larger sample size and investigation of the functional significance of the FokI polymorphism in prostate cancer cells are warranted. PMID- 12540500 TI - Examining the effects of false positive lung cancer screening results on subsequent lung cancer screening adherence. AB - OBJECTIVE: The study goal was to examine the effects of an initial false positive chest X-ray screening result on subsequent lung cancer screening adherence. METHODS: Adherence rates among 4705 individuals in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial at the Henry Ford Health System site with an abnormal/suspicious chest X-ray screening result in the first study year that was subsequently determined to be noncancerous (false positive result, n = 1137 exams) were compared with adherence rates among individuals with an initial negative chest X-ray screening result (n = 3568 exams). RESULTS: Univariate results showed a >50% increase in subsequent nonadherence among individuals with false positive screening results compared with those with negative screening results (17.2% versus 10.3% nonadherence rate, respectively; P < 0.001). Multivariable results showed that statistically significant predictors of nonadherence were false positive cases with current smoking status (P < 0.001) and false positive cases with past smoking status (P < 0.001). Additional predictors of subsequent nonadherence were being African-American (P < 0.01), being female (P < 0.001), and having a high school education or less (P < 0.01). CONCLUSION: Our results demonstrate that the impact of previous screening results, smoking status, race, gender, and education on subsequent screening adherence needs to be weighed carefully, particularly for smokers, an at-risk group, when conducting lung cancer screening intervention studies and perhaps should be considered in clinical practice as well. PMID- 12540502 TI - Timing of pregnancy and the risk of epithelial ovarian cancer. AB - Recent animal studies suggest that progestagen-induced apoptosis of transformed ovarian surface epithelial cells may underlie the observed protective effect of pregnancy on the risk of ovarian cancer. Assuming that increasing numbers of cells are transformed with advancing age, we postulated that the benefits of pregnancy would be greater for older than younger women and tested this hypothesis in a population-based case-control study. We conducted interviews with 620 parous women, ages 18-79 years, with histologically confirmed incident ovarian cancer and 723 parous controls of the same age. Detailed information was collected on reproductive history, as well as hormonal exposures, smoking, medical history, and other factors. We estimated the relative risk of ovarian cancer associated with births at different ages through multiple logistic regression models. After adjusting for parity, older age at first and last births, and shorter time since last birth were all associated with significantly reduced risks of ovarian cancer. Age at first birth and time since last birth were not associated with ovarian cancer when adjusted for each other, whereas age at last birth remained strongly protective [odds ratio (OR), 0.57; 95% confidence interval (CI), 0.36-0.90] among women >35 years versus women less than 25 years. The effect was independent of total parity (per year of age among women with one birth: OR, 0.93; 95%CI, 0.87-1.01; among women with four or more births: OR, 0.96; 95%CI, 0.90-1.02). Our finding that ovarian cancer risk is reduced by pregnancy at older ages is further evidence that pregnancy confers a benefit beyond anovulation and is consistent with the theory that ovarian surface epithelial cell apoptosis induced by pregnancy hormones may be the underlying protective mechanism. PMID- 12540501 TI - Fenretinide breast cancer prevention trial: drug and retinol plasma levels in relation to age and disease outcome. AB - OBJECTIVES: To assess, in women participating in a breast cancer prevention trialon fenretinide (4-HPR), the relationship of drug and retinol levels with the risk of second breast malignancy, taking into account age and menopausal status. METHODS: In a multicenter prevention trial, women with early breast cancer were randomly assigned to receive no treatment or 200 mg of 4-HPR/day for 5 years. Blood was collected at baseline and on a yearly basis during intervention from women recruited at the Istituto Tumori (Milan, Italy; 818 and 756 in the 4-HPR and control arm, respectively, who accounted for 53% of the participants in the trial). The plasma concentrations of 4-HPR, its main metabolite N-(4 methoxyphenyl) retinamide, and retinol were assayed by high-performance liquid chromatography. Three age ranges (or=56 years), menopausal status at baseline, and disease outcome at a median follow-up of 97 months were taken into account in the analysis. RESULTS: Baseline retinol levels were significantly lower (P or=46 years versus or= 0.71; P C (exon 1) in a wobble position. The clinical impact of this polymorphism was investigated in a case-control study in which 312 patients with histologically verified bladder cancer were compared with 254 hospital controls; 13.5% of the cases but only 7.1% of controls were homozygous for the 81C-variant of this polymorphism [odds ratio (OR), 2.04; 95% confidence interval (CI), 1.15-3.61; P = 0.014]. The homozygous 81C genotype was overrepresented, particularly in the patient groups with poorly differentiated tumors (n = 145, >or=G3; OR, 2.22; 95% CI, 1.15-4.27; P = 0.017), muscle-invasive tumors (n = 107, >or=T(2); OR, 2.65; 95% CI, 1.35-5.23; P = 0.005), and flat transitional cell carcinoma (n = 45; OR, 3.69; 95% CI, 1.60 8.51; P = 0.002). In general, 81CC occurred more frequently in advanced types of bladder cancer. We conclude that individuals harboring the homozygous 81C genotype of the H-ras proto-oncogene are at an increased risk of bladder cancer. PMID- 12540508 TI - Correspondence re: Zheng et al, Haplotype of two variants in p16 (CDKN2/MTS 1/INK4a) exon 3 and risk of squamous cell carcinoma of the head and neck: a case control study. 11: 640-645, 2002. PMID- 12540510 TI - Correspondence re: Yasui et al, Breast cancer risk and "delayed" primary Epstein Barr virus infection. 10: 9-16, 2001. PMID- 12540513 TI - Correspondence re: Gammon et al, Environmental toxins and breast cancer on Long Island. I. Polycyclic aromatic hydrocarbon DNA adducts. 11, 677-685, 2002. PMID- 12540514 TI - Correspondence re: Duffield-Lillico et al., Baseline characteristics and the effect of selenium supplementation on cancer incidence in a randomized clinical trial: a summary report of the Nutritional Prevention of Cancer Trial. 11: 630 639, 2002. PMID- 12540516 TI - Receptor gene polymorphisms: lessons on functional relevance from the beta 1 adrenoceptor. PMID- 12540517 TI - Myocardial ischaemia/reperfusion injury and preconditioning: effects of hypercholesterolaemia/hyperlipidaemia. PMID- 12540518 TI - The Rho-kinase inhibitor Y-27632 and the soluble guanylyl cyclase activator BAY41 2272 relax rabbit vaginal wall and clitoral corpus cavernosum. AB - The effects of Y-27632, a Rho-kinase inhibitor and BAY41-2272, a soluble guanylyl cyclase activator, on the tone and nitrergic responses of rabbit vaginal wall and clitoral corpus cavernosum were investigated. Y-27632 and BAY41-2272 (10 nM-10 micro M) elicited concentration-dependent relaxation of phenylephrine-induced tone in both tissues. IC(50) values of Y-27632 for vaginal and clitoral tissues were 370+/-30 nM, and 467+/-14 nM, respectively. BAY41-2272 had IC(50) values of 478+/-54 nM and 304+/-38 nM respectively. The effect of the Y-27632 on the tissue tone was not affected by an inhibitor of nitric oxide synthase (L-NAME; 500 micro M). However, L-NAME reduced the potency of BAY41-2272 in the clitoral corpus cavernosum but not in the vaginal wall. BAY41-2272 enhanced nitrergic relaxation responses only in the clitoral corpus cavernosum. Y-27632 had no effect on nitrergic relaxations in either tissue. These results demonstrate that Y-27632 and BAY41-2272 elicit relaxation of the rabbit vaginal wall and clitoral corpus cavernosum. PMID- 12540519 TI - Attenuation of increased myocardial ischaemia-reperfusion injury conferred by hypercholesterolaemia through pharmacological inhibition of the caspase-1 cascade. AB - 1. Hypercholesterolaemia has been shown to be associated with greater myocardial ischaemia-reperfusion injury, in which apoptosis and inflammation-mediated necrosis both play a key role. 2. Caspase-1 is involved in the activation of both apoptosis and inflammation, through the intermediate of interleukin-1beta (IL 1beta). We herein examined whether pharmacological inhibition of the caspase-1 cascade, using Ac-Tyr-Val-Ala-Asp-CH(2)Cl (Ac-YVAD.cmk), after myocardial ischaemia have greater protective effects on myocardial ischaemia-reperfusion injury in diet-induced hypercholesterolaemic rabbits. 3. Male rabbits fed with standard chow or chow supplemented with 0.5% cholesterol and 10% coconut oil for 8 weeks were subjected to 30 min of left circumflex artery occlusion followed by 4 h of reperfusion. An intravenous bolus of Ac-YVAD.cmk (1.6 mg kg(-1)) or vehicle was given 20 min after coronary occlusion. 4. Postischaemic administration of Ac-YVAD.cmk markedly decreased infarct size from 26+/-3% to 12+/-2% in normally fed rabbits (P=0.005) and from 41+/-6% to 14+/-2% in cholesterol-fed rabbits (P<0.001). 5. In the ischaemic non-necrotic area, treatment with Ac-YVAD.cmk markedly reduced the percentage of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL) positive cardiomyocytes from 15.5+/-0.8% to 2.2+/-0.1% in normally fed rabbits (P<0.001) and from 39.0+/-2.3% to 2.2+/-0.1% in cholesterol-fed rabbits (P<0.001). 6. Ac-YVAD.cmk treatment resulted in a reduction not only of IL-1beta and caspase-1, but also of caspase-3 in the ischaemic myocardium in both normally fed and cholesterol-fed rabbits. 7. No differences in infarct size, the percentage of TUNEL-positive cardiomyocytes, IL-1beta levels or activity of caspase-1 and caspase-3 were observed between Ac-YVAD.cmk-treated normally fed and cholesterol-fed rabbits. 8. This study demonstrates that injection of a selective caspase-1 inhibitor after myocardial ischaemia markedly reduced the detrimental effect conferred by hypercholesterolaemia on myocardial ischaemia reperfusion injury by attenuating both necrotic as well as apoptotic cell death pathways through inhibition of IL-1beta production and activation of caspase-1 and caspase-3. PMID- 12540520 TI - Effects of the AT1-receptor antagonist eprosartan on the progression of left ventricular dysfunction in dogs with heart failure. AB - 1. We examined the effects of eprosartan, an AT(1) receptor antagonist, on the progression of left ventricular (LV) dysfunction and remodelling in dogs with heart failure (HF) produced by intracoronary microembolizations (LV ejection fraction, EF 30 to 40%). 2. Dogs were randomized to 3 months of oral therapy with low-dose eprosartan (600 mg once daily, n=8), high-dose eprosartan (1200 mg once daily, n=8), or placebo (n=8). 3. In the placebo group, LV end-diastolic (EDV) and end-systolic (ESV) volumes increased after 3 months (68+/-7 vs 82+/-9 ml, P<0.004, 43+/-1 vs 58+/-7 ml, P<0.003, respectively), and EF decreased (37+/-1 vs 29+/-1%, P<0.001). In dogs treated with low-dose eprosartan, EF, EDV, and ESV remained unchanged over the course of therapy, whereas in dogs treated with high dose eprosartan, EF increased (38+/-1 vs 42+/-1%, P<0.004) and ESV decreased (41+/-1 vs 37+/-1 ml, P<0.006), Eprosartan also decreased interstitial fibrosis and cardiomyocyte hypertrophy. 4. We conclude that eprosartan prevents progressive LV dysfunction and attenuates progressive LV remodelling in dogs with moderate HF and may be useful in treating patients with chronic HF. PMID- 12540521 TI - Inhibition of kinin breakdown prolongs retention and action of bradykinin in a myocardial B2 receptor compartment. AB - 1. The high efficacy of ACE inhibitors to potentiate the actions of kinins might be explained by a hypothetical compartment in which B(2)-receptors are colocalized with kinin degrading enzymes. To demonstrate the functional consequence of such a compartment we compared the myocardial uptake and the persistence of action of bradykinin under the influence of kininase inhibitors. 2. Bradykinin-induced vasodilation and uptake of tritiated bradykinin were studied in perfused rat hearts during inhibition of ACE and aminopeptidase P. B(2)-receptors were localized by immuno-gold labelling and electron-microscopy. 3. The EC(50) of bradykinin-induced vasodilation (5.1+/-0.8 nM) was shifted to 14 fold lower concentrations during inhibition of both kininases. The maximum persistence of vasodilation after termination of bradykinin application (half life 112+/-20 s) was increased by kininase inhibitors to 398+/-130 s. This prolongation was reversed when B(2)-receptors were blocked simultaneously with the termination of bradykinin infusion. 4. Tritiated bradykinin (perfused for 1 min) was partially (1.7+/-0.24%) retained by the myocardium and consecutively released with a half-life of 70+/-9 s. Kinin uptake was increased during kininase inhibition (7.7+/-2.6%), and was normalized by HOE 140 (2.0+/-0.34%), or when a tritiated B(2)-receptor antagonist (NPC 17731) was used as label. 5. B(2) receptors were localized in plasmalemmal and cytosolic vesicles of capillary endothelium. 6. Bradykinin is locally incorporated and can associate with B(2) receptors repeatedly when kinin breakdown is inhibited. This is the kinetic and functional consequence of a colocalization of kininases and B(2)-receptors in a compartment constituted by endothelial membrane vesicles. PMID- 12540522 TI - Characterization of opioid receptors that modulate nociceptive neurotransmission in the trigeminocervical complex. AB - 1. Opioid agonists have been used for many years to treat all forms of headache, including migraine. We sought to characterize opioid receptors involved in craniovascular nociceptive pathways by in vivo microiontophoresis of micro receptor agonists and antagonists onto neurons in the trigeminocervical complex of the cat. 2. Cats were anaesthetized with alpha-chloralose 60 mg kg(-1), i.p. and 20 mg kg(-1), i.v. supplements after induction and surgical preparation using halothane. Units were identified in the trigeminocervical complex responding to supramaximal electrical stimulation of the superior sagittal sinus, and extracellular recordings of activity made. 3. Seven- or nine-barrelled glass micropipettes incorporating tungsten recording electrodes in their centre barrels were used for microiontophoresis of test substances onto cell bodies. 4. Superior sagittal sinus (SSS)-linked cells whose firing was evoked by microiontophoretic application of L-glutamate (n=8 cells) were reversibly inhibited by microiontophoresis of H(2)N-Tyr-D-Ala-Gly-N-Me-Phe-Gly-ol (DAMGO) (n=12), a selective micro -receptor agonist, in a dose dependent manner, but not by control ejection of sodium or chloride ions from a barrel containing saline. 5. The inhibition by DAMGO of SSS-linked neurons activated with L-glutamate could be antagonized by microiontophoresis of selective micro -receptor antagonists D-Phe Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH(2) (CTOP) or D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr NH(2) (CTAP), or both, in all cells tested (n=4 and 6, respectively). 6. Local iontophoresis of DAMGO during stimulation of the superior sagittal sinus resulted in a reduction in SSS-evoked activity. This effect was substantially reversed 10 min after cessation of iontophoresis. The effect of DAMGO was markedly inhibited by co-iontophoresis of CTAP. 7. Thus, we found that micro -receptors modulate nociceptive input to the trigeminocervical complex. Characterizing the sub-types of opioid receptors that influence trigeminovascular nociceptive transmission is an important component to understanding the pharmacology of this synapse, which is pivotal in primary neurovascular headache. PMID- 12540523 TI - The role of the 8-18 helix of CGRP8-37 in mediating high affinity binding to CGRP receptors; coulombic and steric interactions. AB - 1. The role of individual residues in the 8-18 helix of CGRP(8-37) in promoting high-affinity binding to CGRP(1) receptors expressed on rat L6 and human SK-N-MC cells has been examined. The relative potencies of various derivatives were estimated from their ability to inhibit the human alphaCGRP-mediated increase in cyclic AMP production and the binding of [(125)I]-human alphaCGRP. 2. Arg(11) and Arg(18) were replaced by serines to give [Ser(11,18)]CGRP(8-37). These bound with pKi values <6 to SK-N-MC cells and had apparent pA(2) values of 5.81+/-0.04 and 5.31+/-0.11 on SK-N-MC and L6 cells. CGRP(8-37) had a pKi of 8.22 on SK-N-MC cells and pK(b) values on the above cell lines of 8.95+/-0.04 and 8.76+/-0.04. 3. The arginines were replaced with glutamic acid residues. [Glu(11)]CGRP(8-37) had a pK(b) of 7.14+/-0.14 on SK-N-MC cells (pKi=7.05+/-0.05) and 6.99+/-0.08 on L6 cells. [Glu(18)]CGRP(8-37) had a pK(b) of 7.10+/-0.0.08 on SK-N-MC cells (pKi=6.91+/-0.23) and 7.12+/-0.09 on L6 cells. 4. Leu(12), Leu(15) and Leu(16) were replaced by benzoyl-phenylalanine (bpa) residues. On SK-N-MC cells, the apparent pA(2) values of [bpa(12)]-, [bpa(15)]- and [bpa(16)]CGRP(8-37) were respectively 7.43+/-0.23, 8.34+/-0.11 and 5.66+/-0.16 (pKi values of 7.14+/-0.17, 7.66+/-0.21 and <6): on L6 cells they were 7.96+/-0.36, 8.28+/-0.21 and 6.09+/ 0.04 (all n=3). 5. It is concluded that the Arg(11) and Arg(18) are involved in specific electrostatic interactions with other residues, either on the CGRP(1) receptors or elsewhere on CGRP(8-37). Leu(16) is in a conformationally restricted site when CGRP(8-37) binds to CGRP(1) receptors, unlike Leu(12) and Leu(15). PMID- 12540524 TI - Isolation and pharmacological characterization of a phospholipase A2 myotoxin from the venom of the Irian Jayan death adder (Acanthophis rugosus). AB - 1. It has long been thought that death adder venoms are devoid of myotoxic activity based on studies done on Acanthophis antarcticus (Common death adder) venom. However, a recent clinical study reported rhabdomyolysis in patients following death adder envenomations, in Papua New Guinea, by a species thought to be different to A. antarcticus. Consequently, the present study examined A. rugosus (Irian Jayan death adder) venom for myotoxicity, and isolated the first myotoxin (acanmyotoxin-1) from a death adder venom. 2. A. rugosus (10-50 micro g ml(-1)) and acanmyotoxin-1 (MW 13811; 0.1-1 micro M) were screened for myotoxicity using the chick directly (0.1 Hz, 2 ms, supramaximal V) stimulated biventer cervicis nerve-muscle (CBCNM) preparation. A significant contracture of skeletal muscle and/or inhibition of direct twitches were considered signs of myotoxicity. This was confirmed by histological examination. 3. High phospholipase A(2) (PLA(2)) activity was detected in both A. rugosus venom (140.2+/-10.4 micro mol min(-1) mg(-1); n=6) and acanmyotoxin-1 (153.4+/-11 micro mol min(-1) mg(-1); n=6). Both A. rugosus venom (10-50 micro g ml(-1)) and acanmyotoxin-1 (0.1-1 micro M) caused dose-dependent inhibition of direct twitches and increase in baseline tension (n=4-6). In addition, dose-dependent morphological changes in skeletal muscle were observed. 4. Prior incubation (10 min) of CSL death adder antivenom (5 units ml(-1); n=4) or inactivation of PLA(2) activity with 4-bromophenacyl bromide (1.8 mM; n=4) prevented the myotoxicity caused by acanmyotoxin-1 (1 micro M). 5. Acanmyotoxin-1 (0.1 micro M; n=4) displayed no significant neurotoxicity when it was examined using the indirectly (0.1 Hz, 0.2 ms, supramaximal V) stimulated CBCNM preparation. 6. In conclusion, clinicians may need to be mindful of possible myotoxicity following death adder envenomation in Irian Jaya. PMID- 12540525 TI - Attenuation of the P2Y receptor-mediated control of neuronal Ca2+ channels in PC12 cells by antithrombotic drugs. AB - 1. In PC12 cells, adenine nucleotides inhibit voltage-activated Ca(2+) currents and adenylyl cyclase activity, and the latter effect was reported to involve P2Y(12) receptors. To investigate whether these two effects are mediated by one P2Y receptor subtype, we used the antithrombotic agents 2-methylthio-AMP (2 MeSAMP) and N(6)-(2-methyl-thioethyl)-2-(3,3,3-trifluoropropylthio)-beta,gamma dichloromethylene-ATP (AR-C69931MX). 2. ADP reduced A(2A) receptor-dependent cyclic AMP synthesis with half maximal effects at 0.1-0.17 micro M. In the presence of 30 micro M 2-MeSAMP or 100 nM AR-C69931MX, concentration response curves were shifted to the right by factors of 39 and 30, indicative of pA(2) values of 6.1 and 8.5, respectively. 3. The inhibition of Ca(2+) currents by ADP was attenuated by 10-1000 nM AR-C69931MX and by 3-300 micro M 2-MeSAMP. ADP reinhibited Ca(2+) currents after removal of 2-MeSAMP within less than 15 s, but required 2 min to do so after removal of AR-C69931MX. 4. ADP inhibited Ca(2+) currents with half maximal effects at 5-20 micro M. AR-C69931MX (10-100 nM) displaced concentration response curves to the right, and the resulting Schild plot showed a slope of 1.09 and an estimated pK(B) value of 8.7. Similarly, 10 100 micro M 2-MeSAMP also caused rightward shifts resulting in a Schild plot with a slope of 0.95 and an estimated pK(B) of 5.4. 5. The inhibition of Ca(2+) currents by 2-methylthio-ADP and ADPbetaS was also antagonized by AR-C69931MX, which (at 30 nM) caused a rightward shift of the concentration response curve for ADPbetaS by a factor of 3.8, indicative of a pA(2) value of 8.1. 6. These results show that antithrombotic drugs antagonize the inhibition of neuronal Ca(2+) channels by adenine nucleotides, which suggests that this effect is mediated by P2Y(12) receptors. PMID- 12540526 TI - Bradykinin increases intracellular calcium levels in rat testis peritubular cells via the B2 receptor subtype. AB - 1. RT-PCR and Western blots were used to detect bradykinin B(2) receptors in testis and isolated peritubular cells of pre-pubertal rats. RT-PCR demonstrated expression of a single transcript, whereas Western blots showed up to three specific bands that were in accordance with the described native, glycosylated and dimeric form of B(2) receptor proteins, respectively. 2. Fura-2-loaded peritubular cells responded with an instantaneous, linear and transient rise in [Ca(2+)](i) after adding bradykinin. Stimulation of cells with bradykinin concentrations between 1 micro M and 1 pM showed a dose dependent increase of [Ca(2+)](i). The calcium response to bradykinin was diminished after stimulation of peritubular cells in calcium-free buffer. After blocking the SERCA-pumps by thapsigargin and subsequent stimulation with bradykinin, no rise of [Ca(2+)](i) was appreciated. 3. Multiple stimulation of a single peritubular cell by local perfusion with a brief addition of BK (10 nM) resulted in a fast and immediate response. However, the second and third stimuli had slower rise rates and diminished [Ca(2+)](i) peaks, showing desensitization of the kinin receptor. 4. Addition of the bradykinin B(1) receptor agonist [des-Arg(9)]-bradykinin (100 nM) to Fura-2-loaded peritubular cells did not change the [Ca(2+)](i). However, the B(2) receptor antagonist HOE 140 (100 nM) strongly inhibited the bradykinin induced calcium response. 5. We conclude that the bradykinin-induced increase in [Ca(2+)](i), in testicular peritubular cells is mediated by the stimulation of kinin receptors of the B(2) subtype. PMID- 12540527 TI - Functional characterization of alpha 1-adrenoceptor subtypes in vascular tissues using different experimental approaches: a comparative study. AB - 1. The alpha(1)-adrenergic responses of rat aorta and tail artery have been analysed measuring the contractility and the inositol phosphate (IP) formation induced by noradrenaline. Three antagonists, prazosin, 5-methylurapidil (alpha(1A) selective) and BMY 7378 (alpha(1D) selective) have been used in different experimental procedures. 2. Noradrenaline possesses a greater potency inducing contraction and IP accumulation in aorta (pEC(50)-contraction=7.32+/ 0.04; pEC(50)-IPs=6.03+/-0.08) than in the tail artery (pEC(50) contraction=5.71+/-0.07; pEC(50)-IPs=5.51+/-0.10). Although the maximum contraction was similar in both tissues (E(max)-tail=619.1+/-55.6 mg; E(max) aorta-698.2+/-40.8 mg), there were marked differences in the ability of these tissues to generate intracellular second messengers the tail artery being more efficient (E(max)-tail=1060+/-147%; E(max)-aorta=108.1+/-16.9%). 3. Concentration response curves of noradrenaline in presence of antagonist together with concentration inhibition curves for antagonists added before (CICb) or after (CICa) noradrenaline-induced maximal response in Ca(2+)-containing or Ca(2+)-free medium have been performed. A comparative analysis of the different procedures as well as the mathematical approaches used in each case to calculate the antagonist potencies, were completed. 4. The CICa was the simplest method to characterize the predominant alpha(1)-adrenoceptor subtype involved in the functional response of a tissue. 5. In aorta, where constitutively active alpha(1D)-adrenoeptors are present, the use of different experimental procedures evidenced a complex equilibrium between alpha(1D)- and alpha(1A)-adrenoceptor subtypes. 6. The appropriate management of LiCl in IP accumulation studies allowed us to reproduce the different experimental procedures performed in contractile experiments giving more technical possibilities to this methodology. PMID- 12540528 TI - Tacrolimus and sirolimus decrease oxidative phosphorylation of isolated rat kidney mitochondria. AB - 1. Tacrolimus and sirolimus are potent immunosuppressors used in transplantation. Tacrolimus has been suspected to alter mitochondrial respiration of different tissues but sirolimus has not been evaluated. 2. We evaluated the in vitro effect of tacrolimus and sirolimus on oxidative phosphorylation of isolated rat kidney mitochondria. 3. Oxygen consumption was measured with a Clark-type electrode. Tacrolimus and sirolimus increased the resting rate (state 4) and had no significant effect on ADP-stimulated respiration (state 3). The decrease of respiratory control ratio was concentration-dependent with a biphasic curve for tacrolimus. The EC(50)s were 3.4 x 10(-11) M and 2.3 x 10(-8) M for tacrolimus and 4.4 x 10(-10) M for sirolimus. The maximal inhibition was 20 and 14% for tacrolimus and sirolimus, respectively. 4. Tacrolimus and sirolimus had an uncoupling effect on oxidative phosphorylation related to a decrease of the inner membrane fluidity. At the opposite of cyclosporin A, no effect on swelling or Ca(2+) fluxes was observed. 5. All events occurred at therapeutic concentrations and then could appear during long-term treatment. Cellular consequences such as chronic nephrotoxicity with tacrolimus are suggested. The risk of cyclosporin A nephrotoxicity potentiation by sirolimus is discussed. PMID- 12540529 TI - Alprostadil suppresses angiogenesis in vitro and in vivo in the murine Matrigel plug assay. AB - 1. Prostaglandin E(1) (PGE(1), alprostadil) is used as a vasodilator for the treatment of peripheral vascular diseases. 2. Previous reports suggested a pro angiogenic effect for PGE(1). 3. We studied the in vitro and in vivo effect of PGE(1), complexed with alpha-cyclodextrin, on the angiogenic process. Contrary to what was expected, we found that, in human umbilical vein endothelial cells (HUVECs), PGE(1) inhibited proliferation, migration and capillary-like structure formation in Matrigel. 4. By RT-PCR studies, the expression of the EP(2) and EP(3) subtypes of the PG receptor was detected in HUVECs. 5. PGE(1) alone stimulated adenylate cyclase activity at micromolar concentrations, while at nanomolar concentrations potentiated the forskolin-induced cAMP accumulation. 6. 8-Bromoadenosine-3':5'-cyclic monophosphate (Br-cAMP) mimicked the inhibitory effect of PGE(1) on endothelial cell growth, motility and tube formation. 7. Sulprostone, an agonist at the EP(3) subtype of PG receptors, mimicked the in vitro anti-angiogenic effects of PGE(1), while butaprost, an EP(2) receptor agonist, had no effect. 8. Finally, in the plug assay model of angiogenesis in mice, PGE(1) showed a strong inhibitory effect on Matrigel neovascularization. 9. Thus, PGE(1) possesses strong anti-angiogenic activity in vitro and in vivo. PMID- 12540530 TI - Greater inotropic and cyclic AMP responses evoked by noradrenaline through Arg389 beta 1-adrenoceptors versus Gly389 beta 1-adrenoceptors in isolated human atrial myocardium. AB - 1. We studied the biochemical and contractile responses of isolated human myocardial tissue expressing native receptor variants of the 389G>R beta(1) adrenoceptor polymorphism. 2. Right atrial appendage was obtained from homozygous RR patients (n=37) and homozygous GG patients (n=17) undergoing elective cardiac surgery. The positive inotropic effect of noradrenaline in these tissues, mediated through beta(1)-adrenoceptors, was studied using electrically stimulated (1 Hz) atrial strips, as well as the effects of noradrenaline on cyclic AMP levels and cyclic AMP-dependent protein kinase. 3. Tissue from RR homozygotes (n=14) showed significantly increased inotropic potency to noradrenaline (-log EC(50), M=6.92+/-0.12) compared to GG homozygotes (n=8, -log EC(50), M=6.36+/ 0.11, P<0.005). This difference was not dependent on tissue basal force. 4. Tissue cyclic AMP levels (pmol mg(-1)) were also greater in RR homozygotes (basal 34.8+/-3.7 n=12, 300 nM noradrenaline 41.4+/-7.6 n=9, 30 micro M noradrenaline 45.2+/-3.2 n=22, 0.2 mM isoprenaline 48.3+/-4.2 n=16) compared to GG homozygotes (basal 30.7+/-4.4 n=5, 300 nM noradrenaline 32.6+/-6.92 n=5, 30 micro M noradrenaline 38.1+/-3.1 n=8, 0.2 mM isoprenaline 42.6+/-5.2 n=6, P=0.007). There were no differences between the variants in terms of cyclic AMP-dependent protein kinase activity. 5. These data provide the first evidence that enhanced G-protein coupling of the R389 beta(1)-adrenoceptor variant reported in rodent fibroblast expression systems is also present in native human receptors. The functional consequence of this is to significantly alter the inotropic potency of beta(1) adrenoceptor activation depending on its genotype at the 389 position. PMID- 12540531 TI - Pharmacological characterization of a 1,4-dihydropyridine analogue, 9-(3,4 dichlorophenyl)-3,3,6,6-tetramethyl-3,4,6,7,9,10-hexahydro-1,8(2H,5H) acridinedione (A-184209) as a novelK(ATP) channel inhibitor. AB - 1. This study reports on the identification and characterization of a 1,4 dihydropyridine analogue, 9-(3,4-dichlorophenyl)-3,3,6,6-tetramethyl-3,4,6,7,9,10 hexahydro-1,8(2H,5H)-acridinedione (A-184209) as a novel inhibitor of ATP sensitive K(+) channels. 2. A-184209 inhibited membrane potential changes evoked by the prototypical cyanoguanidine ATP-sensitive K(+) channel opener (KCO) P1075 in both vascular (A10) and urinary bladder smooth muscle cells with IC(50) values of 1.44 and 2.24 micro M respectively. 3. P1075-evoked relaxation of 25 mM K(+) stimulated aortic strips was inhibited by A-184209 in an apparently competitive fashion with a pA(2) value of 6.34. 4. The potencies of A-184209 to inhibit P1075 evoked decreases in membrane potential responses in cardiac myocytes (IC(50)=0.53 micro M) and to inhibit 2-deoxyglucose-evoked cation efflux pancreatic RINm5F cells (IC(50)=0.52 micro M) were comparable to the values for inhibition of smooth muscle K(ATP) channels. 5. On the other hand, a structural analogue of A 184209 that lacked the gem-dimethyl substituent, 9-(3,4-dichlorophenyl) 3,4,6,7,9,10-hexahydro-1,8(2H,5H)-acridinedione (A-184208), was found to be a K(ATP) channel opener, evoking membrane potential responses in A10 smooth muscle cells (EC(50)=385 nM) and relaxing aortic smooth muscle strips (IC(50)=101 nM) in a glyburide-sensitive manner. 6. Radioligand binding studies demonstrated that A 184209 displaced SUR1 binding defined by [(3)H]glyburide binding to RINm5F cell membranes with a K(i) value of 0.11 micro M whereas A-184208 was ineffective. On the other hand, both A-184209 (K(i)=1.34 micro M) and A-184208 (K(i)=1.14 micro M) displaced binding of the KCO radioligand, [(125)I]A-312110 in guinea-pig bladder membranes with similar affinities. 7. These studies demonstrate that A 184209 is a novel and structurally distinct compound that inhibits K(ATP) channels in smooth muscle with potencies comparable to glyburide. The structural overlap between DHP openers and blockers, together with their differential interaction with ligand binding sites, support the notion that both openers and blockers bind to similar or very closely coupled sites on the sulfonylurea receptor and that subtle changes in the pharmacophore itself could switch functional properties from K(ATP) channel activation to inhibition. PMID- 12540532 TI - Coupling of the nucleotide P2Y4 receptor to neuronal ion channels. AB - 1. G protein-linked P2Y nucleotide receptors are known commonly to stimulate the phosphoinositide signalling pathway. However, we have previously demonstrated that the cloned P2Y(2), P2Y(6) and P2Y(1) receptors couple to neuronal N-type Ca(2+) channels and to M-type K(+) channels. Here we investigate the coupling of recombinant, neuronally expressed rat- and human P2Y(4) receptors (rP2Y(4), hP2Y(4)) to those channels. 2. Rat sympathetic neurones were nuclear-injected with a P2Y(4) cDNA plasmid. A subsequent activation of rP2Y(4) or hP2Y(4) by UTP (100 micro M) in whole-cell (ruptured-patch) mode produced only about 12% inhibition of the N-type Ca(2+) current (I(Ca(N))). Surprisingly, in perforated patch mode, UTP produced much more inhibition of I(Ca(N)) (maximally 51%), with an IC(50) value of 273 nM. This inhibition was voltage-dependent and was blocked by co-expression of the betagamma-binding transducin Galpha-subunit. Pertussis toxin (PTX) pretreatment also suppressed I(Ca(N)) inhibition. 3. UTP inhibited the M-current, recorded in perforated patch mode, by (maximally) 52%, with IC(50) values of 21 nM for rP2Y(4) and 28 nM for hP2Y(4). This inhibition was not affected by PTX pretreatment. 4. With rP2Y(4), ATP inhibited the M-current (IC(50) 524 nM, 26 times weaker than UTP), whereas ATP had no agonist activity at hP2Y(4). This suggests a difference in agonist binding site between rP2Y(4) and hP2Y(4). 5. We conclude that, in contrast to other nucleotide receptors studied, the P2Y(4) receptor couples much more effectively to M-type K(+) channels than to Ca(2+) channels. Coupling to the Ca(2+) channels involves the betagamma-subunits of G(i/o)-proteins and requires a diffusible intracellular component that is lost in ruptured-patch recording. PMID- 12540533 TI - Understanding naturally acquired immunity to Plasmodium falciparum malaria. PMID- 12540534 TI - Archaea and their potential role in human disease. PMID- 12540535 TI - Plasmodium falciparum infection elicits both variant-specific and cross-reactive antibodies against variant surface antigens. AB - Naturally acquired antibodies to Plasmodium falciparum erythrocyte membrane protein-1 (PfEMP-1), the variant surface antigens expressed on the surface of infected erythrocytes, are thought to play a role in protection against P. falciparum malaria. Here, we have studied the development of antibodies to PfEMP 1 in adult malaria patients living in Rourkela, India, an area with a low malaria transmission rate, and prevalence of antibodies to PfEMP-1 in residents of San Dulakudar, India, a village in which P. falciparum malaria is hyperendemic. Convalescent-phase sera from adult malaria patients from Rourkela agglutinate homologous P. falciparum isolates as well as some heterologous isolates, suggesting that they develop partially cross-reactive antibodies to PfEMP-1 following infection. Adult sera from San Dulakudar agglutinate diverse P. falciparum isolates, suggesting that they have antibodies with wide recognition of diverse PfEMP-1. Mixed-agglutination assays using pairs of P. falciparum isolates confirm the presence of both variant-specific and partially cross reactive antibodies in convalescent-phase sera from Rourkela and adult sera from San Dulakudar. Analysis of PfEMP-1 sequences suggests a molecular basis for the observed cross-reactivity. PMID- 12540536 TI - Human eosinophil peroxidase induces surface alteration, killing, and lysis of Mycobacterium tuberculosis. AB - The antimycobacterial role of eosinophil peroxidase (EPO), one of the most abundant granule proteins in human eosinophils, was investigated. Our data indicate that purified EPO shows significant inhibitory activity towards Mycobacterium tuberculosis H37Rv. On a molar basis, this activity was similar to that exhibited by neutrophil myeloperoxidase (MPO) and was both dose and time dependent. In contrast to the activity of MPO, which requires H(2)O(2), EPO also exhibited anti-M. tuberculosis activity in the absence of exogenously added peroxide. Morphological evidence confirmed that the mechanism of action of EPO against mycobacteria differs from that of MPO. While MPO kills M. tuberculosis H37Rv exclusively in the presence of hydrogen peroxide, it does not induce morphological changes in the pathogen. In contrast, EPO-treated bacteria frequently had cell wall lesions and eventually underwent lysis, either in the presence or in the absence of H(2)O(2). PMID- 12540537 TI - Induction of proinflammatory cytokines in human lung epithelial cells during Chlamydia pneumoniae infection. AB - Chlamydia pneumoniae is an obligate intracellular human pathogen that causes acute respiratory diseases such as pneumonia and bronchitis. Previous studies have established that C. pneumoniae can induce cytokines in mouse and/or human cells, but little information is available on the cytokine response of respiratory epithelial cells, a first line of infection. In this study, heparin treatment of C. pneumoniae significantly reduced its ability to induce interleukin 8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) mRNA in human lung carcinoma cells, indicating that cytadherence is an important early stimulus for induction of proinflammatory mediators. Although the IL-8, gamma interferon, and TNF-alpha message was consistently induced by infection of A549 cells not treated with heparin, only an elevation of IL-8 protein was detected in A549 supernatants. A549 IL-beta and IL-6 mRNA and supernatant protein profiles were not significantly changed by infection. Heat or UV inactivation of C. pneumoniae only partially reduced the cytokine response, and inhibition of C. pneumoniae protein or DNA synthesis did not affect its ability to induce cytokine gene expression. To prevent stress-induced cytokine release by the A549 cells, centrifugation was not utilized for infection experiments. These experiments establish the importance of cytadherence in cytokine release by cells of respiratory epithelial origin and suggest that further work in the area of cytokine mediators is warranted to gain valuable pathogenic and therapeutic insights. PMID- 12540538 TI - Nramp1 is not a major determinant in the control of Brucella melitensis infection in mice. AB - Brucella, the causative agent of brucellosis in animals and humans, can survive and proliferate within macrophages. Macrophages mediate mouse resistance to various pathogens through the expression of the Nramp1 gene. The role of this gene in the control of Brucella infection was investigated. When BALB/c mice (Nramp1(s)) and C.CB congenic mice (Nramp1(r)) were infected with Brucella melitensis, the number of Brucella organisms per spleen was significantly larger in the C.CB mice than in the BALB/c mice during the first week postinfection (p.i.). This Nramp1-linked susceptibility to Brucella was temporary, since similar numbers of Brucella were recovered from the two strains of mice 2 weeks p.i. The effect of Nramp1 expression occurred within splenocytes intracellularly infected by BRUCELLA: However, there was no difference between in vitro replication rates of Brucella in macrophages isolated from the two strains of mice infected in vivo or in Nramp1 RAW264 transfectants. In mice, infection with Brucella induced an inflammatory response, resulting in splenomegaly and recruitment of phagocytes in the spleen, which was amplified in C.CB mice. Reverse transcription-PCR (RT-PCR), performed 5 days p.i., showed that inducible nitric oxide synthase, tumor necrosis factor alpha (TNF-alpha), interleukin-12 p40 (IL-12p40), gamma interferon (IFN-gamma), and IL-10 mRNAs were similarly induced in spleens of the two strains. In contrast, the mRNA of KC, a C-X-C chemokine, was induced only in infected C.CB mice at this time. This pattern of mRNA expression was maintained at 14 days p.i., with IFN-gamma and IL-12p40 mRNAs being more intensively induced in the infected C.CB mice, but TNF-alpha mRNA was no longer induced. The higher recruitment of neutrophils observed in the spleens of infected C.CB mice could explain the temporary susceptibility of C.CB mice to B. melitensis infection. In contrast to infections with Salmonella, Leishmania, and Mycobacterium, the expression of the Nramp1 gene appears to be of limited importance for the natural resistance of mice to Brucella. PMID- 12540539 TI - Molecular and phenotypic analysis of the CS54 island of Salmonella enterica serotype typhimurium: identification of intestinal colonization and persistence determinants. AB - The shdA gene is carried on a 25-kb genetic island at centisome 54 (CS54 island) of the Salmonella enterica serotype Typhimurium chromosome. In addition to shdA, the CS54 island of Salmonella serotype Typhimurium strain LT2 contains four open reading frames designated ratA, ratB, sivI, and sivH. DNA hybridization analysis revealed that the CS54 island is comprised of two regions with distinct phylogenetic distribution within the genus Salmonella. Homologues of shdA and ratB were detected only in serotypes of Salmonella enterica subsp. I. In contrast, sequences hybridizing with ratA, sivI, and sivH were present in S. enterica subsp. II and S. bongori in addition to S. enterica subsp. I. Deletion of the ratA and sivI genes did not alter the ability of Salmonella serotype Typhimurium to colonize the organs of mice. Insertional inactivation of the sivH gene resulted in defective colonization of the Peyer's patches of the terminal ileum but normal colonization of the cecum, mesenteric lymph nodes, and spleen. Deletion of the shdA gene resulted in decreased colonization of the cecum and Peyer's patches of the terminal ileum and colonization to a lesser degree in the mesenteric lymph nodes and spleen 5 days post-oral inoculation of mice. A strain containing a deletion in the ratB gene exhibited a defect for the colonization of the cecum but not of the Peyer's patches, mesenteric lymph nodes, and spleen. The shdA and ratB deletion strains exhibited a shedding defect in mice, whereas the sivH deletion strain was shed at numbers similar to the wild type. These data suggest that colonization of the murine cecum is required for efficient fecal shedding in mice. PMID- 12540540 TI - Clostridium sordellii phospholipase C: gene cloning and comparison of enzymatic and biological activities with those of Clostridium perfringens and Clostridium bifermentans phospholipase C. AB - The gene encoding Clostridium sordellii phospholipase C (Csp) was cloned and expressed as a histidine-tagged (His-tag) protein, and the protein was purified to compare its enzymatic and biological activities with those of Clostridium perfringens phospholipase C (Cpa) and Clostridium bifermentans phospholipase C (Cbp). Csp was found to consist of 371 amino acid residues in the mature form and to be more homologous to Cbp than to Cpa. The egg yolk phospholipid hydrolysis activity of the His-tag Csp was about one-third of that of His-tag Cpa, but the hemolytic activity was less than 1% of that of His-tag Cpa. His-tag Csp was nontoxic to mice. Immunization of mice with His-tag Cbp or His-tag Csp did not provide effective protection against the lethal activity of His-tag Cpa. These results indicate that Csp possesses similar molecular properties to Cbp and suggest that comparative analysis of toxic and nontoxic clostridial phospholipases is helpful for characterization of the toxic properties of clostridial phospholipases. PMID- 12540541 TI - The msbB mutant of Neisseria meningitidis strain NMB has a defect in lipooligosaccharide assembly and transport to the outer membrane. AB - A deletion-insertion mutation in msbB, a gene that encodes a lipid A acyltransferase, was introduced into encapsulated Neisseria meningitidis serogroup B strain NMB and an acapsular mutant of the same strain. These mutants were designated NMBA11K3 and NMBA11K3cap-, respectively. Neither lipooligosaccharide (LOS) nor lipid A could be isolated from NMBA11K3 although a number of techniques were tried, but both were easily extracted from NMBA11K3cap . Immunoelectron microscopy using monoclonal antibody (MAb) 6B4, which recognizes the terminal Galbeta1-4GlcNAc of LOS, demonstrated that NMB, NMBcap-, and NMBA11K3cap- expressed LOS circumferentially, while MAb 6B4 did not bind to the surface of NMBA11K3. However, cytoplasmic staining of NMBA11K3 with MAb 6B4 was a consistent observation. Mass-spectrometric analyses demonstrated that the relative amounts of the lipid A-specific C12:0 3-OH and C14:0 3-OH present in the membrane preparations (MP) from NMBA11K3 were substantially decreased (25- and 23 fold, respectively) compared to the amount in MP from its parent strain, NMB. Western blot analyses of MP from NMBA11K3 demonstrated that the levels of porin in the outer membrane of NMBA11K3 were also substantially decreased. These studies suggest that the lipid A acylation defect in encapsulated NMBA11K3 influences the assembly of the lipid A and consequently the incorporation of porin in the outer membrane. PMID- 12540542 TI - Availability of complement bound to Staphylococcus aureus to interact with membrane complement receptors influences efficiency of phagocytosis. AB - Complement-mediated opsonization of encapsulated Staphylococcus aureus (CP+) of the predominant capsule types, 5 and 8, remains poorly understood. Our previous work showed that complement is important for mouse survival of CP+ type 5 bacteremia and that the type 5 capsule inhibits the binding of opsonic C3 fragments to the organism. The importance of complement-mediated opsonization of CP+ was tested by neutrophil phagocytosis assays. Complement-mediated opsonization of CP+ increased phagocytosis by 57% compared to opsonization in complement-inhibited serum. Agar-grown CP+, enhancing capsule expression, was phagocytosed only one-tenth as well as the capsule-negative organisms (CP-), supporting the belief that staphylococcal polysaccharide capsules impair phagocytosis. Despite relatively poor phagocytosis of CP+ compared to CP-, complement activation increased the phagocytosis of CP+ by 103%. Thus, complement in normal human serum may have an important role in opsonizing CP+, even when capsule expression is strong. The ability of bound C3 fragments to interact with complement receptor 1 (CD35) on the membrane of human erythrocytes was tested in an immune adherence assay. S. aureus capsule was able to mask C3 fragments on the organism from binding to complement receptor 1. The inhibition of C3 binding to CP+ and the masking of deposited C3 fragments caused by the presence of capsule was associated with markedly decreased phagocytosis. The addition of anti-capsule antibodies to normal human serum was found to markedly improve the recognition of deposited C3 fragments by complement receptor 1 even when the absolute number of C3 molecules bound to S. aureus was not increased. PMID- 12540544 TI - Restoration of cytoadherence to an adherence-deficient mutant of Mycoplasma arthritidis by genetic complementation. AB - Mycoplasma arthritidis causes a severe septic arthritis in rats under natural and experimental conditions. An earlier study implicated a membrane lipoprotein designated MAA1 in cytadherence of M. arthritidis. In addition, a spontaneous adherence-deficient mutant was shown to contain a nonsense mutation in the gene encoding MAA1, resulting in production of a truncated product, MAA1Delta. In the present study, a wild-type maa1 gene carried on transposon Tn4001T was introduced into the low-adherence mutant by polyethylene glycol-mediated transformation. The presence of the tranposon and the wild-type maa1 gene in the chromosome of transformants was confirmed by PCR and Southern hybridization. The latter procedure also confirmed that each transformant contained a single copy of the transposon. Western immunoblotting showed that transformants produced both wild type MAA1 and MAA1Delta, indicating that the introduced wild-type maa1 gene was functional. This phenotype was stably maintained after multiple subcultures even in the absence of antibiotic selection. Finally, transformants were shown to adhere to rat L-2 lung cells in culture at wild-type levels, providing confirmation for an important role for MAA1 in adherence. PMID- 12540543 TI - Toll-like receptor 4-positive macrophages protect mice from Pasteurella pneumotropica-induced pneumonia. AB - This study investigates Toll-like receptor 4 (TLR4)-positive macrophages in early recognition and clearance of pulmonary bacteria. TLR4 is a trans-membrane receptor that is the primary recognition molecule for lipopolysaccharide of gram negative bacteria. The TLR4(Lps-del) mouse strains C57BL10/ScN (B10) and STOCK Abb(tm1) TLR4(Lps-del) Slc11a1(s)(B10 x C2D) are susceptible to pulmonary infections and develop pneumonia when naturally or experimentally infected by the opportunistic bacterium Pasteurella pneumotropica. Since these mice have the TLR4(Lps-del) genotype, we hypothesized that reconstitution of mice with TLR4 positive macrophages would provide resistance to this bacterium. A cultured macrophage cell line (C2D macrophages) and bone marrow cells from C2D mice were adoptively transferred to B10 and B10 x C2D mice by intraperitoneal injection. C2D macrophages increased B10 and B10 x C2D mouse resistance to P. pneumotropica. In C2D-recipient mice there was earlier transcription of tumor necrosis factor alpha and chemokines JE and macrophage inflammatory protein 2 (MIP-2) in the lungs of B10 and B10 x C2D mice, and there was earlier transcription of KC and MIP-1alpha in B10 x C2D mice. In addition, the course of inflammation following experimental Pasteurella challenge was altered in C2D recipients. C2D macrophages also protected B10 x C2D mice, which lack CD4(+) T cells. These data indicate that macrophages are critical for pulmonary immunity and can provide host resistance to P. pneumotropica. This study indicates that TLR4-positive macrophages are important for early recognition and clearance of pulmonary bacterial infections. PMID- 12540545 TI - Roles of sortase in surface expression of the major protein adhesin P1, saliva induced aggregation and adherence, and cariogenicity of Streptococcus mutans. AB - Sortase is a newly discovered transpeptidase that covalently links LPXTGX containing surface proteins to the gram-positive bacterial cell wall. In this study, the sortase gene (srtA) was isolated from Streptococcus mutans NG8 by PCR. The gene encoded a 246-amino-acid protein, including a 40-amino-acid signal peptide. The srtA gene was insertionally inactivated by a tetracycline resistance cassette. P1, a major surface protein adhesin previously shown to anchor to the peptidoglycan by the LPXTGX motif, was secreted into the culture medium by the srtA mutant. In contrast, the wild-type P1 remained cell wall associated. Complementation of the mutant with srtA restored the P1 surface expression phenotype. P1 produced by the mutant, but not that produced by the wild type and the srtA-complemented mutant, was recognized by an antibody raised against the hydrophobic domain and charged tail C terminal to the LPXTGX motif. These results suggest that the failure to anchor P1 to the cell wall is due to the lack of cleavage of P1 at the LPXTGX motif. The srtA mutant was markedly less hydrophobic than the wild type and the complemented mutant. The srtA mutant failed to aggregate in the presence of saliva or salivary agglutinin and adhered poorly to saliva- or salivary agglutinin-coated hydroxylapatite. In rats, the srtA mutant colonized the teeth poorly when sucrose was absent. When sucrose was present, the srtA mutant colonized the teeth but less effectively and induced significantly less caries (P < 0.05) than the wild-type strain. In conclusion, the sortase enzyme in S. mutans is responsible for anchoring P1 to the cell surface and plays a role in modulating the surface properties and cariogenicity of S. mutans. PMID- 12540546 TI - A/J mice are susceptible and C57BL/6 mice are resistant to Listeria monocytogenes infection by intragastric inoculation. AB - Previous studies demonstrated that the innate resistance of mice to Listeria monocytogenes infection by intravenous or intraperitoneal inoculation is regulated principally by the Hc locus on mouse chromosome 2. The A/J and C57BL/6 mouse strains were identified as prototype L. monocytogenes-susceptible and resistant strains, respectively. In the present study, we compared the relative susceptibilities of A/J and C57BL/6 mice to intragastric (i.g.) inoculation with L. monocytogenes. The results of our study indicate that A/J mice are significantly more susceptible than C57BL/6 mice to an i.g. challenge with L. monocytogenes. This was reflected in the estimated 50% lethal doses for the two strains (10(6) and 10(8) CFU for A/J and C57BL/6 mice, respectively) and a more rapid and severe dissemination of the infection to the spleen and liver in A/J mice than in C57BL/6 mice. Histopathological examination of tissues from the infected mice confirmed the greater severity of disease in A/J mice. Clearance of a primary infection enhanced the resistance of both A/J and C57BL/6 mice to reinfection with L. monocytogenes via the gastrointestinal tract. However, the relative difference in susceptibility between the two strains was evident even after immunization. The A/J mouse holds promise as a model for investigating the pathogenesis of gastrointestinal listeriosis because of its ability to develop systemic infection following challenge with numbers of organisms similar to those recovered from some L. monocytogenes-contaminated food products. PMID- 12540547 TI - Lon, a stress-induced ATP-dependent protease, is critically important for systemic Salmonella enterica serovar typhimurium infection of mice. AB - Studies on the pathogenesis of Salmonella enterica serovar Typhimurium infections in mice have revealed the presence of two prominent virulence characteristics-the invasion of the nonphagocytic cells to penetrate the intestinal epithelium and the proliferation within host phagocytic cells to cause a systemic spread and the colonization of host organs. We have recently demonstrated that the ATP-dependent Lon protease of S. enterica serovar Typhimurium negatively regulates the efficiency of invasion of epithelial cells and the expression of invasion genes (A. Takaya et al., J. Bacteriol. 184:224-232, 2002). This study was performed to reveal the contribution of the Lon protease to the virulence of S. enterica serovar Typhimurium in mice. Determination of 50% lethal doses for the lon disruption mutant and wild-type strain revealed that the mutant was highly attenuated when administered either orally or intraperitoneally to BALB/c mice. The mutant was also found to be able to reach extraintestinal sites but unable to proliferate efficiently within the spleen and cause lethal systemic disease of mice. Macrophage survival assays revealed that the lon disruption mutant could not survive or proliferate within murine macrophages. In addition, the mutant showed extremely increased susceptibility to hydrogen peroxide, which contributes to the bactericidal capacity of phagocytes. The mutant also showed increased sensitivity to acidic conditions. Taken together, the impaired ability of the lon disruption mutant to survive and grow in macrophages could be due to the enhanced susceptibility to the oxygen-dependent killing mechanism associated with respiratory burst and the low phagosomal pH. These results suggest that the Lon protease is essentially involved in the systemic infection of mice with S. enterica serovar Typhimurium, which can be fatal. Of further interest is the finding that the lon disruption mutant persists in the BALB/c mice for long periods without causing an overwhelming systemic infection. PMID- 12540548 TI - Different innate ability of I/St and A/Sn mice to combat virulent Mycobacterium tuberculosis: phenotypes expressed in lung and extrapulmonary macrophages. AB - Mice of the I/St and A/Sn inbred strains display a severe and moderate course, respectively, of disease caused by Mycobacterium tuberculosis. Earlier, we showed that the response to mycobacterial antigens in I/St mice compared to that in A/Sn mice is shifted toward Th2-like reactivity and a higher proliferative activity and turnover of T cells. However, the physiologic basis for different expressions of tuberculosis severity in these mice remains largely unknown. Here, we extend our previous observations with evidence that I/St interstitial lung macrophages are defective in the ability to inhibit mycobacterial growth and to survive following in vitro infection with M. tuberculosis H37Rv. A unique feature of this phenotype is its exclusive expression in freshly isolated lung macrophages. The defect is not displayed in ex vivo macrophages obtained from the peritoneal cavity nor in macrophages developed in vitro from progenitors extracted from various organs, including the lung itself. In addition, we show that, in sharp contrast to peritoneal macrophages, the mycobactericidal capacity of lung macrophages is not elevated in the presence of exogenous gamma interferon. Our data suggest that the in vivo differentiation in a particular anatomical microenvironment determines the pattern of macrophage-mycobacterium interaction. Thus, caution should be exercised when conclusions based upon the results obtained in a particular in vitro system are generalized to the functions of all phagocytes during M. tuberculosis infection. PMID- 12540549 TI - Mycobacterium bovis BCG vaccines exhibit defects in alanine and serine catabolism. AB - Mycobacterium bovis BCG is the only accepted vaccine for the prevention of tuberculosis (TB) in humans. BCG is a live vaccine, and induction of immunity to TB requires productive infection of the host by BCG. However, BCG is not a satisfactory vaccine, because it fails to protect against pulmonary TB in adults. In this study, we found that BCG strains cannot utilize many naturally occurring amino acids as the sole nitrogen source for growth. This defect is caused, at least partially, by the lack of functional metabolic enzymes. All BCG strains are unable to catabolize L-alanine or D-alanine due to a frameshift mutation in the L alanine dehydrogenase gene (ald). Some BCG strains, such as BCG-Pasteur and BCG Frappier, cannot catabolize L-serine, apparently due to inadequate expression of L-serine deaminase (sdaA). We also found that undegraded alanine and serine inhibit the growth of BCG through blockage of glutamine synthetase. These results suggest that BCG strains are limited in nitrogen metabolic capacity and predict defects that may restrict multiplication and persistence of the live vaccine within the host. PMID- 12540550 TI - Oral treponemes and their outer membrane extracts activate human gingival epithelial cells through toll-like receptor 2. AB - Oral treponemes are considered to be important in the development and progression of periodontal diseases. We investigated the mechanisms of recognition and activation of human gingival epithelial cells (HGEC) with the oral treponemes Treponema denticola, Treponema vincentii, and Treponema medium and their outer membrane extracts (OMEs). T. vincentii and T. medium but not T. denticola produced interleukin 8 (IL-8) in an HGEC culture. Further, all three treponemes induced IL-8 mRNA expression and NF-kappaB activation in HGEC. Among them, T. denticola especially exhibited trypsin- and chymotrypsin-like protease activities, and the addition of chymostatin, a chymotrypsin protease inhibitor, resulted in detectable IL-8 production by HGEC cultured with T. denticola. Additionally, IL-8 mRNA expression in HGEC cultured with the three treponemes and their OMEs was definitely inhibited by the mouse anti-human Toll-like receptor 2 (TLR2) monoclonal antibody TL2.1. These findings suggest that oral treponemes and their OMEs activate HGEC through TLR2. PMID- 12540552 TI - Role of systemic and mucosal immune responses in reciprocal protection against Bordetella pertussis and Bordetella parapertussis in a murine model of respiratory infection. AB - The roles of systemic humoral immunity, cell-mediated immunity, and mucosal immunity in reciprocal protective immunity against Bordetella pertussis and Bordetella parapertussis were examined by using a murine model of respiratory infection. Passive immunization with serum from mice infected with B. pertussis established protective immunity against B. pertussis but not against B. parapertussis. Protection against B. parapertussis was induced in mice that had been injected with serum from mice infected with B. parapertussis but not from mice infected with B. pertussis. Adoptive transfer of spleen cells from mice infected with B. pertussis or B. parapertussis also failed to confer reciprocal protection. To examine the role of mucosal immunity in reciprocal protection, mice were infected with preparations of either B. pertussis or B. parapertussis, each of which had been incubated with the bronchoalveolar wash of mice that were convalescing after infection with B. pertussis or B. parapertussis. Such incubation conferred reciprocal protection against B. pertussis and B. parapertussis on infected mice. The data suggest that mucosal immunity including secreted immunoglobulin A in the lungs might play an important role in reciprocal protective immunity in this murine model of respiratory infection. PMID- 12540551 TI - Protective levels of diphtheria-neutralizing antibody induced in healthy volunteers by unilateral priming-boosting intranasal immunization associated with restricted ipsilateral mucosal secretory immunoglobulin a. AB - Subunit intranasal vaccines offer the prospect of inducing combined systemic mucosal immunity against mucosally transmitted infections such as human immunodeficiency virus. However, although human studies have demonstrated the induction of active immunity, secretory immunoglobulin A (sIgA) responses are variable, and no study has demonstrated protection by accepted vaccine-licensing criteria as measured by direct toxin-neutralizing activity. Using the genetically inactivated mutant diphtheria toxoid CRM(197) in a bioadhesive polycationic polysaccharide chitosan delivery system, we found that a single nasal immunization was well tolerated and boosted antitoxin neutralizing activity in healthy volunteers, which could be further boosted by a second immunization. The neutralizing activity far exceeded accepted protective levels and was equivalent to that induced by standard intramuscular vaccine and significantly greater than intranasal immunization with CRM(197) in the absence of chitosan. A striking but unexpected observation was that although unilateral intranasal immunization induced circulating antitoxin antibody-secreting cells, a nasal antitoxin sIgA response was seen only after the second immunization and only in the vaccinated nostril. If these data are reproduced in larger studies, an intranasal diphtheria vaccine based on CRM(197)-chitosan could be rapidly licensed for human use. However, a restricted sIgA response suggests that care must be taken in the priming-boosting strategy and clinical sampling techniques when evaluating such vaccines for the induction of local mucosal immunity. PMID- 12540553 TI - Sensitivity of polarized epithelial cells to the pore-forming toxin aerolysin. AB - Aerolysin is one of the major virulence factors produced by Aeromonas hydrophila, a human pathogen that produces deep wound infection and gastroenteritis. The toxin interacts with target mammalian cells by binding to the glycan core of glycosylphosphatidyl inositol (GPI)-anchored proteins and subsequently forms a pore in the plasma membrane. Since epithelial cells of the intestine are the primary targets of aerolysin, we investigated its effect on three types of polarized epithelial cells: Caco-2 cells, derived from human intestine; MDCK cells, a well-characterized cell line in terms of protein targeting; and FRT cells, an unusual cell line in that it targets its GPI-anchored proteins to the basolateral plasma membrane in contrast to other epithelial cells, which target them almost exclusively to the apical surface. Surprisingly, we found that all three cell types were sensitive to the toxin from both the apical and the basolateral sides. Apical sensitivity was always higher, even for FRT cells. In contrast, FRT cells were more sensitive from the basolateral than from the apical side to the related toxin Clostridium septicum alpha-toxin, which also binds to GPI-anchored proteins but lacks the lectin binding domain found in aerolysin. These observations are consistent with the notion that a shuttling mechanism involving low-affinity interactions with surface sugars allows aerolysin to gradually move toward the membrane surface, where it can finally encounter the glycan cores of GPI-anchored proteins. PMID- 12540554 TI - Elevated immunoglobulin E against recombinant Brugia malayi gamma-glutamyl transpeptidase in patients with bancroftian filariasis: association with tropical pulmonary eosinophilia or putative immunity. AB - A major allergen of the lymphatic filarial nematode Brugia malayi, a homologue of gamma-glutamyl transpeptidase (gamma-GT), is involved in the pathology of tropical pulmonary eosinophilia (TPE) through its potent allergenicity and the induction of antibodies against the host pulmonary epithelium. To investigate the immunoglobulin G (IgG) subclass and IgE responses to recombinant B. malayi gamma GT, we analyzed the results obtained from 51 patients with differing clinical manifestations of bancroftian filariasis. gamma-GT-specific IgG1, rather than IgG4, was the predominant IgG subclass, particularly in patients with TPE (geomean, 6,321 ng/ml; range, 78 to 354,867 ng/ml) and was 75 times higher than in patients with elephantiasis (CP) (P < 0.003) and 185 times higher than in endemic normal individuals (ENL) (P < 0.010). IgG2 responses were low and IgG3 was almost absent, with no significant differences among the groups. gamma-GT specific IgG4 responses were significantly elevated in those with subclinical microfilaremia (MF) compared to the CP and ENL groups and correlated with the presence of circulating filarial antigen (CAg). More significantly, gamma-GT specific IgE antibody levels were strikingly elevated in patients with TPE (geomean, 681 ng/ml; range, 61 to 23,841 ng/ml) and in the ENL group (geomean, 106 ng/ml; range, 13 to 1,405 ng/ml) whereas the gamma-GT-specific IgE level was 44 and 61 times lower in those with MF and CP, respectively (P < 0.001). Elevated gamma-GT-specific IgE/IgG4 ratios were demonstrated in patients with TPE (ratio, 45) and ENL (ratio, 107). Because expression of gamma-GT in Brugia infective third-stage larvae (L3) was demonstrated by immunoblot analysis, the elevated gamma-GT-specific IgE antibodies appear to be associated not only with pulmonary pathology but also with possible resistance to infection in lymphatic filariasis. PMID- 12540555 TI - Characterization of the conformational epitope of Guy's 13, a monoclonal antibody that prevents Streptococcus mutans colonization in humans. AB - Guy's 13 is a mouse monoclonal antibody which recognizes streptococcal antigen I/II (SA I/II), a major cell surface glycoprotein of Streptococcus mutans. In a number of clinical trials, this antibody has been shown to prevent colonization in the human oral cavity. The aim of this study was to identify the SA I/II epitope recognized by Guy's 13. The data suggest that the epitope is conformational, delimited by two noncontiguous regions of the antigen: residues 45 to 457, within the N-terminal half of SA I/II, and residues 816 to 983, within the C-terminal half. In fluid-phase immunoassays a strict requirement for the simultaneous presence of both regions was demonstrated for antibody binding. Furthermore, these two regions of SA I/II were shown to have the ability to interact with each other in the absence of Guy's 13 antibody, suggesting that the normal conformation of SA I/II might be determined by the interaction of these two regions. PMID- 12540556 TI - Saccharomyces boulardii interferes with enterohemorrhagic Escherichia coli induced signaling pathways in T84 cells. AB - Enterohemorrhagic Escherichia coli (EHEC) infections are associated with the modification of tight-junction permeability and synthesis of proinflammatory cytokine interleukin-8 (IL-8). In a previous study, it was demonstrated that EHEC induced IL-8 secretion is due to the involvement of the mitogen-activated protein kinase (MAPK), AP-1, and NF-kappaB pathways. In this study, we investigated the effect of the yeast Saccharomyces boulardii on EHEC infection in T84 cells. For this purpose, cells were (i) incubated with bacteria and yeast at the same time or (ii) incubated overnight with yeast cells that were maintained during infection or eliminated by several washes before infection. Coincubation is sufficient to maintain the transmonolayer electrical resistance (TER) of EHEC infected cells, whereas the preincubation of cells with the yeast without elimination of the yeast during infection is necessary to significantly decrease IL-8 secretion. We thus analyzed the mechanisms of S. boulardii action. We showed that S. boulardii has no effect on EHEC growth or on EHEC adhesion. Kinetics studies revealed that EHEC-induced myosin light chain (MLC) phosphorylation precedes the decrease of TER. ML-7, an MLC kinase inhibitor, abolishes the EHEC induced MLC phosphorylation and decrease of TER. Studies show that S. boulardii also abolishes EHEC-induced MLC phosphorylation. We demonstrated that the preincubation of cells with S. boulardii without washes before EHEC infection inhibits NF-kappaB DNA binding activity, phosphorylation and degradation of IkappaB-alpha, and activation of the three members of a MAPK group (extracellular signal-regulated protein kinases 1 and 2, p38, and c-jun N-terminal kinase). These findings demonstrate that S. boulardii exerts a preventive effect on EHEC infection by (i) interfering with one of the transduction pathways implicated in the control of tight-junction structure and (ii) decreasing IL-8 proinflammatory secretion via inhibition of the NF-kappaB and MAPK signaling pathways in infected T84 cells. PMID- 12540557 TI - Heterogeneity of mycolactones produced by clinical isolates of Mycobacterium ulcerans: implications for virulence. AB - Mycobacterium ulcerans is the causative agent of Buruli ulcer, a severe necrotizing skin disease endemic in tropical countries. Clinical evidence suggests that M. ulcerans isolates from Asia, Mexico, and Australia may be less virulent than isolates from Africa. In vivo studies suggest that mycolactone, a polyketide-derived macrolide toxin, plays a major role in the tissue destruction and immune suppression which occur in cases of Buruli ulcer. Mycolactones were extracted from 34 isolates of M. ulcerans representing strains from Africa, Malaysia, Asia, Australia, and Mexico. Thin-layer chromatography, mass spectroscopic analysis, and cytopathic assays of partially purified mycolactones from these isolates revealed that M. ulcerans produces a heterogeneous mixture of mycolactone variants. Mycolactone A/B, the most biologically active mycolactone species, was identified by mass spectroscopy as [M(+)Na](+) at m/z 765.5 in all cytotoxic isolates except for those from Mexico. Mycolactone C [M+Na](+) at m/z 726.3 was the dominant mycolactone species in eight Australian isolates, and mycolactone D [M+Na](+) m/z 781.2 was characteristic of two Asian strains. Mycolactone species are conserved within specific geographic areas, suggesting that there may be a correlation between mycolactone profile and virulence. In addition, the core lactone, [M+Na](+) m/z 447.4, was identified as a minor species, supporting the hypothesis that mycolactones are synthesized by two polyketide synthases. A cytopathic assay of the core lactone showed that this molecule is sufficient for cytotoxicity, although it is much less potent than the complete mycolactone. PMID- 12540558 TI - Streptococcus pyogenes glycoprotein-binding strepadhesin activity is mediated by a surface-associated carbohydrate-degrading enzyme, pullulanase. AB - The interactions between pathogenic bacteria and the host need to be resolved at the molecular level in order to develop novel antiadhesive drugs and vaccines. We have previously identified strepadhesin, a novel glycoprotein-binding activity in Streptococcus pyogenes binding to thyroglobulin, submaxillar mucin, fetuin, and asialofetuin. The activity is known to be regulated by Mga, a regulator of streptococcal virulence factors, and is carried by the surface-associated streptococcal cysteine protease, SpeB. In the present study, we focused on the high strepadhesin activity in an S. pyogenes strain (NZ131rgg) lacking SpeB expression. By extracting surface proteins from the bacteria, a new strepadhesin protein was identified, and mass spectrometric analysis and database search identified it as a putative pullulanase. The gene was cloned, and the recombinant pullulanase (PulA) exhibited pullulanase and starch hydrolyzing activity, as well as strepadhesin activity. Sequencing of the pulA gene revealed an open reading frame with 3,498 bp encoding a protein of 1,165 amino acids with a predicted molecular mass of 129 kDa. PulA exhibited properties typical for a gram-positive surface protein with a putative signal sequence and LPKTGE cell wall anchoring motif and contained the four highly conserved regions common to pullulanases. Mutant bacteria deficient in PulA expression showed diminished strepadhesin activity on bacterial dot blot assay and reduced adherence to thyroglobulin immobilized on microtiter plates. Thus, S. pyogenes strepadhesin activity is carried by a surface-bound pullulanase, which combines glycoprotein-binding and carbohydrate-degrading activities in the same molecule. PMID- 12540559 TI - Colonization of C57BL/6J and BALB/c wild-type and knockout mice with Helicobacter pylori: effect of vaccination and implications for innate and acquired immunity. AB - The gram-negative bacterial pathogen Helicobacter pylori is a major cause of peptic ulcer disease and a risk factor for gastric cancer in humans. Adapted H. pylori strains, such as strain SS1, are able to infect mice and are a useful model for gastric colonization and vaccination studies. In this study we used a streptomycin-resistant derivative of H. pylori SS1 to analyze the colonization behavior and the success of vaccination in wild-type (wt) and various knockout mice of the BALB/c and C57BL/6J genetic backgrounds. We here report that BALB/c interleukin-4 knockout (IL-4(-/-)) mice are weakly overcolonized compared to the wt strain but that the IL-12(-/-) knockout results in a strong overcolonization (500%). Unexpectedly, in the C57BL/6J background the same knockouts behaved in diametrically opposed manners. The IL-4(-/-) mutation caused a 50% reduction and the IL-12(-/-) knockout caused a 95% reduction compared to the wt colonization rate. For C57BL/6J mice we further analyzed the IL-18(-/-) and Toll-like receptor 2 knockout mutations, which showed reductions to 66 and 57%, respectively, whereas mice with the IL-10(-/-) phenotype were hardly infected at all (5%). In contrast, the tumor necrosis factor receptor knockout (p55(-/-) and p55/75(-/-)) mice showed an overcolonization compared to the C57BL/6J wt strain. With exception of the low-level infected C57BL/6J IL-10(-/-) and IL-12(-/-) knockout mice, all knockout mutants were accessible to a prophylactic vaccination and their vaccination behavior was comparable to that of the wt strains. PMID- 12540560 TI - Use of a promoter trap system in Bacillus anthracis and Bacillus subtilis for the development of recombinant protective antigen-based vaccines. AB - We have recently reported Bacillus anthracis attenuated live vaccine strains efficiently expressing recombinant protective antigen (rPA) and have shown a direct correlation between the level of rPA secreted by these cells and efficacy (S. Cohen, I. Mendelson, Z. Altboum, D. Kobiler, E. Elhanany, T. Bino, M. Leitner, I. Inbar, H. Rosenberg, Y. Gozes, R. Barak, M. Fisher, C. Kronman, B. Velan, and A. Shafferman, Infect. Immun. 68:4549-4558, 2000). To isolate more potent Bacillus promoters for a further increase in the production of rPA, we developed a promoter trap system based on various gfp reporter genes adapted for use in both Bacillus subtilis and B. anthracis backgrounds. Accordingly, a B. anthracis library of 6,000 clones harboring plasmids with chromosomal B. anthracis DNA fragments inserted upstream from gfpuv was constructed. Based on fluorescence intensity, 57 clones carrying potentially strong promoters were identified, some of which were DNA sequenced. The most potent B. anthracis promoter identified (Pntr; 271 bp) was 500 times more potent than the native pagA promoter and 70 times more potent than the alpha-amylase promoter (Pamy). This very potent promoter was tested along with the other promoters (which are three, six, and eight times more potent than Pamy) for the ability to drive expression of rPA in either B. subtilis or B. anthracis. The number of cell-associated pre PA molecules in B. anthracis was found to correlate well with the strength of the promoter. However, there appeared to be an upper limit to the amount of mature PA secreted into the medium, which did not exceed that driven by Pamy. Furthermore, the rPA constructs fused to the very potent promoters proved to be deleterious to the bacterial hosts and consequently led to genetic instability of the PA expression plasmid. Immunization with attenuated B. anthracis expressing rPA under the control of promoters more potent than Pamy was less efficient in eliciting anti-PA antibodies than that attained with Pamy. The results are consistent with the notion that overexpression of PA leads to severe secretion stress and have practical implications for the design of second-generation rPA based vaccines. PMID- 12540561 TI - Five-member gene family of Bartonella quintana. AB - Bartonella quintana, the agent of trench fever and an etiologic agent of bacillary angiomatosis, has an extraordinarily high hemin requirement for growth compared to other bacterial pathogens. We previously identified the major hemin receptor of the pathogen as a 30-kDa surface protein, termed HbpA. This report describes four additional homologues that share approximately 48% amino acid sequence identity with hbpA. Three of the genes form a paralagous cluster, termed hbpCAB, whereas the other members, hbpD and hbpE, are unlinked. Secondary structure predictions and other evidence suggest that Hbp family members are beta barrels located in the outer membrane and contain eight transmembrane domains plus four extracellular loops. Homologs from a variety of gram-negative pathogens were identified, including Bartonella henselae Pap31, Brucella Omp31, Agrobacterium tumefaciens Omp25, and neisserial opacity proteins (Opa). Family members expressed in vitro-synthesized proteins ranging from ca. 26.5 to 35.1 kDa, with the exception of HbpB, an approximately 55.9-kDa protein whose respective gene has been disrupted by a approximately 510 GC-rich element containing variable-number tandem repeats. Transcription analysis by quantitative reverse transcriptase-PCR (RT-PCR) indicates that all family members are expressed under normal culture conditions, with hbpD and hbpB transcripts being the most abundant and the rarest, respectively. Mutagenesis of hbpA by allelic exchange produced a strain that exhibited an enhanced hemin-binding phenotype relative to the parental strain, and analysis by quantitative RT-PCR showed elevated transcript levels for the other hbp family members, suggesting that compensatory expression occurs. PMID- 12540562 TI - Does host complement kill Borrelia burgdorferi within ticks? AB - The Lyme disease spirochete, Borrelia burgdorferi, inhabits the gut lumen of the tick vector. At this location the spirochete is exposed to host blood when a tick feeds. We report here on studies that were done with normal and complement deficient (C3-knockout) mice to determine if the host complement system killed spirochetes within the vector. We found that spirochete numbers within feeding nymphs were not influenced by complement, most likely because host complement was inactivated within the vector. The Lyme disease outer surface protein A (OspA) vaccine is a transmission-blocking vaccine that targets spirochetes in the vector. In experiments with mice hyperimmunized with OspA, complement was not required to kill spirochetes within nymphs and to block transmission from nymphs to the vaccinated host. However, host complement did enhance the ability of OspA antibody to block larvae from acquiring spirochetes. Thus, the effects of OspA antibody on nymphal transmission and larval acquisition appear to be based on different mechanisms. PMID- 12540563 TI - Effect of inhibition of extracellular signal-regulated kinase 1 and 2 pathway on apoptosis and bcl-2 expression in Helicobacter pylori-infected AGS cells. AB - Helicobacter pylori induces activation of mitogen-activated protein kinases (MAPKs). However, its effect on H. pylori-induced apoptosis has not been evaluated. Thus, we examined whether H. pylori-induced extracellular signal regulated kinase 1 and 2 (ERK1/2) and p38 MAPK activation affects gastric epithelial cell apoptosis and bcl-2 family gene expression, especially in relation to the cagA status of an H. pylori strain. In flow cytometric and oligonucleosome-bound DNA enzyme-linked immunosorbent assay analyses, infection with cagA(+) H. pylori strains induced gastric cancer cell apoptosis in AGS cells more prominently than infection with cagA mutants. Activation of ERK1/2 and p38 MAPKs was also more prominent in cagA(+) strains. Pretreatment with a MEK inhibitor (PD98059) inhibited ERK1/2 activation and increased H. pylori-induced apoptosis significantly. This increased apoptosis was accompanied by decreased antiapoptotic bcl-2 mRNA expression among bcl-2-related genes (bcl-2, bax, bak, mcl-1, and bcl-X(L/S)), and the effect was also more prominent in the cagA(+) strains. However, the alteration of bcl-2 gene expression was not accompanied by protein level changes. Inhibition of p38 using specific inhibitor SB203580 decreased H. pylori-induced apoptosis but resulted in little alteration of bcl-2 related gene expression. In conclusion, H. pylori-induced ERK1/2 activation, especially by the cagA(+) H. pylori strain, may play a protective role against gastric epithelial cell apoptosis partially through maintenance of bcl-2 gene expression. PMID- 12540564 TI - Entamoeba histolytica cysteine proteinases disrupt the polymeric structure of colonic mucin and alter its protective function. AB - The adherent mucous gel layer lining the colonic epithelium is the first line of host defense against invasive pathogens, such as Entamoeba histolytica. The mucous layer prevents the attachment of amoeba to the colonic epithelium by trapping and aiding in the expulsion of the parasite. Disruption of the mucous layer is thought to occur in invasive amebiasis, and the mechanism by which the parasite overcomes this barrier is not known. The aim of this study was to characterize the specific interactions occurring between E. histolytica secreted cysteine proteinases and colonic mucin as a model to examine the initial events of invasive amebiasis. E. histolytica secreted products were examined for mucinase activity utilizing mucin metabolically labeled with [(35)S]cysteine as a substrate. Cysteine proteinases degraded mucin in a time- and dose-dependent manner. A significant reduction (>50%) in high-molecular-weight mucin with altered buoyant density was observed when degraded mucin was analyzed by Sepharose 4B column chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography, and CsCl density gradient centrifugation. Mucinase activity was eliminated by the specific cysteine protease inhibitor trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane and was independent of glycosidase activity. Moreover, the degraded mucin was 38% less effective than native mucin at inhibiting amebic adherence to target epithelial cells. These results are the first to show that E. histolytica cysteine proteinases alter the protective function of the mucous barrier by disrupting the structure of the MUC2 polymer. Mechanistically, the parasite achieves this via proteolytic degradation of the terminal cysteine-rich domains. PMID- 12540565 TI - Induction by sphingomyelinase of shiga toxin receptor and shiga toxin 2 sensitivity in human microvascular endothelial cells. AB - Shiga toxin-producing enterohemorrhagic Escherichia coli is the major cause of acute renal failure in young children. The interaction of Shiga toxins 1 and 2 (Stx1 and Stx2) with endothelial cells is an important step in the renal coagulation and thrombosis observed in hemolytic uremic syndrome. Previous studies have shown that bacterial lipopolysaccharide and host cytokines slowly sensitize endothelial cells to Shiga toxins. In the present study, bacterial neutral sphingomyelinase (SMase) rapidly (1 h) sensitized human dermal microvascular endothelial cells (HDMEC) to the cytotoxic action of Stx2. Exposure of endothelial cells to neutral SMase (0.067 U/ml) caused a rapid increase of intracellular ceramide that persisted for hours. Closely following the change in ceramide level was an increase in the expression of globotriaosylceramide (Gb3), the receptor for Stx2. A rapid increase was also observed in the mRNA for ceramide:glucosyltransferase (CGT), the first of three glycosyltransferase enzymes of the Gb3 biosynthetic pathway. The product of CGT (glucosylceramide) was also increased. In contrast, mRNA for the third enzyme of the pathway, Gb3 synthase, was constitutively produced and was not influenced by SMase treatment of HDMEC. These results describe a rapid response mechanism by which extracellular neutral SMase derived from either bacteria or eukaryotic cells may signal endothelial cells to become sensitive to Shiga toxins. PMID- 12540566 TI - DNA from periodontopathogenic bacteria is immunostimulatory for mouse and human immune cells. AB - Although bacterial DNA (bDNA) containing unmethylated CpG motifs stimulates innate immune cells through Toll-like receptor 9 (TLR-9), its precise role in the pathophysiology of diseases is still equivocal. Here we examined the immunostimulatory effects of DNA extracted from periodontopathogenic bacteria. A major role in the etiology of periodontal diseases has been attributed to Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Peptostreptococcus micros. We therefore isolated DNA from these bacteria and stimulated murine macrophages and human gingival fibroblasts (HGF) in vitro. Furthermore, HEK 293 cells transfected with human TLR-9 were also stimulated with these DNA preparations. We observed that DNA from these pathogens stimulates macrophages and gingival fibroblasts to produce tumor necrosis factor alpha and interleukin-6 in a dose-dependent manner. Methylation of the CpG motifs abolished the observed effects. Activation of HEK 293 cells expressing TLR-9 which were responsive to bDNA but not to lipopolysaccharide confirmed that immunostimulation was achieved by bDNA. In addition, the examined bDNA differed in the ability to stimulate murine macrophages, HGF, and TLR-9-transfected cells. DNA from A. actinomycetemcomitans elicited a potent cytokine response, while DNA from P. gingivalis and P. micros showed lower immunostimulatory activity. Taken together, the results strongly suggest that DNA from A. actinomycetemcomitans, P. gingivalis, and P. micros possesses immunostimulatory properties in regard to cytokine secretion by macrophages and fibroblasts. These stimulatory effects are due to unmethylated CpG motifs within bDNA and differ between distinct periodontopathogenic bacteria strains. Hence, immunostimulation by DNA from A. actinomycetemcomitans, P. gingivalis, and P. micros could contribute to the pathogenesis of periodontal diseases. PMID- 12540567 TI - Protection of chickens against Escherichia coli infections by DNA containing CpG motifs. AB - Synthetic oligodeoxynucleotides (ODN) containing CpG motifs (CpG-ODN) have been shown to be effective immunoprotective agents in murine models for a variety of viral, intracellular bacterial, and protozoan infections. Until now, the use of CpG-ODN to protect against extracellular bacterial infections has not been reported. The objective of this study was to investigate the effect of CpG-ODN against cellulitis and colibacillosis in broiler chickens, using a well established model. At 22 days of age, birds received CpG-ODN by either the subcutaneous or intramuscular route. Three days later, a virulent isolate of Escherichia coli was applied to a scratch site on the caudal abdominal skin. Birds were examined for 10 days after the E. coli challenge, and pathological and bacteriological assessments were conducted on all birds. The control group of birds receiving no CpG-ODN((2007)) had a survival rate of 15%. In contrast, groups that received CpG-ODN((2007)), by either subcutaneous or intramuscular injection, had significantly higher survival rates (P < 0.0001). Furthermore, the size of the cellulitis lesion was significantly smaller in groups that received CpG-ODN((2007)) by the subcutaneous route (P < 0.01). A dose of as little as 3.16 micro g of CpG-ODN((2007)), delivered 3 days prior to challenge by either the subcutaneous or intramuscular route, significantly protected birds against E. coli infection (P < 0.01). This study demonstrates that CpG-ODN((2007)) has both local and systemic protective effects in broiler chickens. This is the first time that CpG-ODN((2007)) has been demonstrated to have an immunoprotective effect against an extracellular bacterial infection in any food animal species. PMID- 12540568 TI - Immunopathogenesis of pulmonary granulomas in the guinea pig after infection with Mycobacterium tuberculosis. AB - Pulmonary tuberculosis in guinea pigs is similar to the disease in humans and is accordingly widely used as a model to test tuberculosis vaccines. The primary site of expression of acquired immunity and the hallmark of tuberculosis is the granuloma. Granuloma morphology is well described, but there is limited information regarding T-cell subset influx. We monitored the course of pulmonary tuberculosis in guinea pigs and observed four distinct immunohistopathological stages. In all stages there were similar numbers and arrangement of CD4 and CD8 T cells. There were only small numbers of apoptotic lymphocytes, scattered around and within the necrotic core, and acid-fast bacilli were visible both within macrophages and free within airway debris. A key finding of the study was the observation that the development of the necrotic core was an early event and almost certainly preceded the emergence of the acquired immune response. This in turn suggests that innate mechanisms are the basis of the early lesions and that subsequent acquired responses are unable to moderate them. This hypothesis differs from the current dogma that excessive activity of T cells mediates delayed-type hypersensitivity and that cellular cytolysis is the root cause of the necrosis. PMID- 12540569 TI - Disruption of epithelial barrier integrity by Salmonella enterica serovar typhimurium requires geranylgeranylated proteins. AB - Epithelial cells that line the human intestinal mucosa constitute the initial sites of host invasion by bacterial pathogens. A number of bacteria, such as Salmonella and Yersinia spp., have been shown to disrupt the integrity of the epithelial barrier, although little is known about the mechanisms underlying that effect. We found that polarized MDCK-1 epithelial cells infected with invasive Salmonella enterica serovar Typhimurium SL1344 exhibited marked changes in F actin organization, an increase in the paracellular flux of dextran, and a rapid decrease in transepithelial electrical resistance (TER). In contrast, infection with an isogenic noninvasive mutant (hilA) increased the TER in these cells. Pretreating MDCK-1 cells with the inhibitors for tyrosine kinase (genistein) or phosphatidylinositol 3-kinase (wortmannin) did not affect invasion and subsequent perturbation of the epithelial barrier by serovar Typhimurium. Instead, the geranylgeranyltransferase 1 inhibitor GGTI-298, but not the farnesyltransferase inhibitor FTI-277, clearly reversed the capacity of serovar Typhimurium to disrupt the epithelial barrier. The substrates for GGTI-298 include Rho family GTPases, as indicated by inhibiting prenylation of Rac1 and Cdc42. Infection with wild-type serovar Typhimurium increased the level of activated Rac1 and Cdc42 and caused these proteins to accumulate apically in MDCK-1 cells. This Salmonella induced accumulation of Rac1 and Cdc42 and alteration of the junction-associated proteins ZO-1, occludin, and E-cadherin in MDCK-1 cells were markedly inhibited by GGTI-298. These results suggest that activation of geranylgeranylated proteins, including Rac1 and Cdc42, is critical for disruption of barrier integrity by serovar Typhimurium in polarized MDCK-1 cells. PMID- 12540570 TI - Direct continuous method for monitoring biofilm infection in a mouse model. AB - We have developed a rapid, continuous method for real-time monitoring of biofilms, both in vitro and in a mouse infection model, through noninvasive imaging of bioluminescent bacteria colonized on Teflon catheters. Two important biofilm-forming bacterial pathogens, Staphylococcus aureus and Pseudomonas aeruginosa, were made bioluminescent by insertion of a complete lux operon. These bacteria produced significant bioluminescent signals for both in vitro studies and the development of an in vivo model, allowing effective real-time assessment of the physiological state of the biofilms. In vitro viable counts and light output were parallel and highly correlated (S. aureus r = 0.98; P. aeruginosa r = 0.99) and could be maintained for 10 days or longer, provided that growth medium was replenished every 12 h. In the murine model, subcutaneous implantation of the catheters (precolonized or postimplant infected) was well tolerated. An infecting dose of 10 (3) to 10 (5) CFU/catheter for S. aureus and P. aeruginosa resulted in a reproducible, localized infection surrounding the catheter that persisted until the termination of the experiment on day 20. Recovery of the bacteria from the catheters of infected animals showed that the bioluminescent signal corresponded to the CFU and that the lux constructs were highly stable even after many days in vivo. Since the metabolic activity of viable cells could be detected directly on the support matrix, nondestructively, and noninvasively, this method is especially appealing for the study of chronic biofilm infections and drug efficacy studies in vivo. PMID- 12540571 TI - Phagocytosis and intracellular fate of Aspergillus fumigatus conidia in alveolar macrophages. AB - Aspergillus fumigatus is the most prevalent airborne fungal pathogen responsible for fatal invasive aspergillosis in immunocompromised patients. Upon arrival in the lung alveolus, conidia of A. fumigatus are phagocytosed by alveolar macrophages, the major phagocytic cells of the lung. Engulfment and intracellular trafficking of A. fumigatus conidia in alveolar macrophages of two different origins, the murine cell line MH-S and human pulmonary alveolar macrophages, were analyzed by electron microscopy and immunofluorescence. Phagocytosis of A. fumigatus conidia required actin polymerization and phosphatidylinositol 3-kinase activity. Fusion of A. fumigatus phagosomes with early and late endosomes was shown by immunolabeling with specific markers for the transferrin receptor, early endosome antigen, and Rab7. Maturation of A. fumigatus phagolysosomes was monitored by using a fixable acidotropic probe, LysoTracker Red DND-99, and an anti-cathepsin D antibody. Bafilomycin A-induced inhibition of lysosomal acidification abolished the conidial killing by the macrophages. These data suggest that the maturation of A. fumigatus phagosomes results from fusion with the compartments of the endocytic pathway and that the killing of conidia depends on phagolysosome acidification. A model for the phagocytosis of A. fumigatus conidia by alveolar macrophages is proposed on the basis of these results. PMID- 12540572 TI - Colonial morphology of Burkholderia cepacia complex genomovar III: implications in exopolysaccharide production, pilus expression, and persistence in the mouse. AB - The purpose of this study was to determine the role of colonial morphology of Burkholderia cepacia complex (BCC) organisms in pathogenicity in a mouse model of pulmonary infection. BCC strain C1394 was rapidly cleared by leukopenic mice after intranasal challenge, whereas a spontaneous variant (C1394mp2) that was indistinguishable from the parent strain by genetic typing persisted in the lungs and differed in colonial morphology. The parent strain had a matte colonial phenotype, made scant exopolysaccharide (EPS), and was lightly piliated. The variant had a shiny phenotype, produced abundant EPS, and was heavily piliated. Matte to shiny colonial transformation was induced by growth at 42 degrees C. Colonial morphology in the BCC strain variant was associated with persistence after pulmonary challenge and appeared to be correlated with the elaboration of putative virulence determinants. PMID- 12540573 TI - Vaccine-induced reduction of Helicobacter pylori colonization in mice is interleukin-12 dependent but gamma interferon and inducible nitric oxide synthase independent. AB - Previous studies with mice have shown that major histocompatibility complex class II (MHC-II) is required for protection from Helicobacter pylori, while MHC-I and antibodies are not. Thus, CD4(+) T cells are presumed to play an essential role in protective immunity via secretion of cytokines. To determine which cytokines are associated with a reduction of bacterial load in immunized mice, gastric cytokine expression was examined by semiquantitative reverse transcription-PCR in protected (defined as > or =2-log-unit decrease in bacterial load) and unprotected mice 4 weeks after challenge. Elevated levels of mRNA for interleukin 12p40 (IL-12p40), gamma interferon (IFN-gamma), tumor necrosis factor alpha, and inducible nitric oxide synthase (iNOS) were associated with protection in immunized-challenged (I/C) mice, but Th2 cytokine (IL-4, IL-5, IL-10, and IL-13) and chemokine (KC, MIP-2, and MCP-1) expression was not associated with protection. Despite the association of IFN-gamma and iNOS message with protection, I/C mice genetically lacking either of these products were able to reduce the bacterial load as well as the wild-type I/C controls. The I/C mice lacking IL-12p40 were not protected compared to unimmunized-challenged mice. All I/C groups developed gastritis. We conclude that neither IFN-gamma nor iNOS is essential for vaccine-induced protection from H. pylori infection. The p40 subunit of IL-12, which is a component of both IL-12 and IL-23, is necessary for protection in immunized mice. These findings suggest a novel IFN-gamma independent function of IL-12p40 in effective mucosal immunization against H. pylori. PMID- 12540574 TI - Transposon mutagenesis of Mycobacterium marinum identifies a locus linking pigmentation and intracellular survival. AB - Pathogenic mycobacteria survive and replicate within host macrophages, but the molecular mechanisms involved in this necessary step in the pathogenesis of infection are not completely understood. Mycobacterium marinum has recently been used as a model for aspects of the pathogenesis of tuberculosis because of its close genetic relationship to Mycobacterium tuberculosis and because of similarities in the pathology and course of infection caused by this organism in its natural hosts, fish and frogs, with tuberculosis in humans. In order to advance the utility of the M. marinum model, we have developed efficient transposon mutagenesis of the organism by using a Drosophila melanogaster mariner based transposon. To determine the efficiency of transposition, we have analyzed pigmentation mutants from the transposon mutant library. In addition to insertions in four known genes in the pathway of pigment biosynthesis, two insertions in novel genes were identified in our mutant library. One of these is in a putative inhibitor of the carotenoid biosynthesis pathway. The second unexpected insertion is in an intergenic region between two genes homologous to Rv2603c and Rv2604c of M. tuberculosis. In addition to a pigmentation defect, this mutant showed increased susceptibility to singlet oxygen and grew poorly in murine macrophages. Complementation with M. tuberculosis genomic DNA encompassing Rv2603c to Rv2606c corrected the pigmentation and growth defects of the mutant. These data demonstrate the utility of mariner-based transposon mutagenesis of M. marinum and that M. marinum can be used to study the function of M. tuberculosis genes involved in intracellular survival and replication. PMID- 12540575 TI - Phagocytic activity and monocyte chemotactic protein expression by pulmonary macrophages in persistent pulmonary cryptococcosis. AB - The mechanisms by which Cryptococcus neoformans persists in an immunocompetent host are not well understood. Using a rat model of persistent infection, we investigated the ability of pulmonary macrophages (PuM) to phagocytize C. neoformans and produce monocyte chemotactic protein 1 (MCP-1) as a function of the length of time of infection and opsonin. The ability of macrophages to affect serum-mediated phagocytosis varied over the course of infection and was dependent on CD11b/c and CD18 expression. Infection resulted in increased MCP-1 levels within the lung, though the actual amounts varied over the course of infection. Immunohistochemical studies localized MCP-1 expression to macrophages and epithelioid cells. Enhanced production of MCP-1 by PuM from infected rats was confirmed by ex vivo studies. Induction of MCP-1 following serum-mediated phagocytosis was observed for PuM from both infected and noninfected rats and depended on the interaction of C. neoformans with CD11b/c and CD18. Specific antibody was more efficient than serum in promoting phagocytosis and consistently elicited more MCP-1. The relative amount of MCP-1 produced in association with phagocytosis was similar for PuM at all lengths of time of infection. Decreased MCP-1 production was observed for PuM obtained from older rats, including long term (8 to 10 months)-infected and age-matched controls, suggesting that aging may affect the production of MCP-1 by PuM in response to cryptococcal infection. In summary, our results show that macrophages are an important source of MCP-1 during pulmonary cryptococcosis and that MCP-1 production is actively regulated during infection. Furthermore, we find that phagocytosis of C. neoformans can serve as an important stimulus for MCP-1 production by PuM, though the efficiency of this process is dependent on the opsonin type and may be affected by aging. PMID- 12540576 TI - Application of a Saccharomyces cerevisiae model to study requirements for trafficking of Yersinia pestis YopM in eucaryotic cells. AB - YopM is a leucine-rich repeat (LRR) virulence protein that is delivered into host cells when any of the three human-pathogenic species of Yersinia binds to mammalian cells. It exhibits heterogeneity of size and sequence among the yersiniae, but the functional consequences of this variability are not yet known. Yersinia pestis YopM was previously shown to accumulate in the nuclei of infected HeLa cells by a mechanism that requires vesicular trafficking. In this study, we characterized the trafficking of Y. pestis YopM in a Saccharomyces cerevisiae model previously found to support nuclear localization of YopM from an enteropathogenic Yersinia strain (C. F. Lesser and S. I. Miller, EMBO J. 20:1840 1849, 2001). Y. pestis YopM was N-terminally fused to the yeast enhanced green fluorescent protein (yEGFP) and inducibly expressed in the cytoplasm. yEGFP-YopM localized to the yeast nucleus, showing that this property is conserved for YopMs so far tested and that infection and the presence of other Yops are not required for its trafficking. When expressed in S. cerevisiae that is temperature sensitive for vesicular transport, YopM failed to accumulate in the nucleus at the nonpermissive temperature but did accumulate when the permissive temperature was restored. This shows that vesicular trafficking also is required in yeast for normal localization of YopM. YopM consists of a 71-residue leader sequence, 15 LRRs, and a 32-residue tail. Deletion analysis revealed that the leader sequence or tail is alone insufficient to direct YopM to the nucleus, showing that the LRR structure is required. Both the N-terminal and C-terminal halves of YopM localized to the nucleus, indicating the possible presence of two nuclear localization signals (NLSs) in YopM or domains in YopM where an NLS-containing protein might bind; this fits with the presence of two highly conserved regions among Yersinia YopMs. yEGFP-YopM lacking LRRs 4 to 7 or 7 to 10 accumulated in the nucleus in yeast, and YopM lacking these LRRs concentrated normally in the HeLa cell nucleus after delivery by Yersinia infection, showing that these LRRs are not essential for YopM trafficking in eucaryotic cells. However, because Y. pestis carrying either of these YopMs is strongly compromised in virulence in mice, these findings revealed that LRRs 4 to 10 map a region of YopM or support a conformation of YopM that is necessary for a pathogenic effect. PMID- 12540577 TI - Infection by Streptococcus pyogenes induces the receptor activator of NF-kappaB ligand expression in mouse osteoblastic cells. AB - Group A Streptococcus pyogenes is known to induce nongonococcal septic arthritis in addition to pharyngitis, scarlet fever, and poststreptococcal sequelae. However, little is known about the interaction between S. pyogenes and bone cells. We report here that S. pyogenes strain JRS4 (M6) attached to and invaded mouse primary osteoblasts. Reverse transcription-PCR demonstrated that S. pyogenes infection of osteoblasts stimulated expression of mRNA for the receptor activator of NF-kappaB ligand (RANKL). Western blot analysis followed by ligand precipitation with the receptor activator of NF-kappaB receptor showed that there was an increase in RANKL protein in infected osteoblasts. Production of interleukin-6 was also stimulated, but no production of interleukin-1beta or tumor necrosis factor alpha was observed. Stimulation of RANKL production was not observed in osteoblasts stimulated with heat-inactivated S. pyogenes, suggesting that an active interaction of S. pyogenes with osteoblasts is essential for this phenomenon. A Western blot analysis performed with antibodies specific for phosphorylated signal transduction proteins demonstrated that S. pyogenes infection induces phosphorylation of p38 mitogen-activated protein kinase. A specific inhibitor of this kinase, SB203580, inhibited RANKL production by infected osteoblasts. These results suggest that infection of osteoblasts by S. pyogenes stimulates RANKL production and may trigger bone destruction in infected bone tissue. PMID- 12540578 TI - Immunohistochemical evaluation of T cells in oral lesions from human immunodeficiency virus-positive persons with oropharyngeal candidiasis. AB - Oropharyngeal candidiasis (OPC), caused by Candida albicans, is the most frequent opportunistic fungal infection in human immunodeficiency virus (HIV)-positive persons. Although Th1-type CD4(+) T cells are considered important for host defense against mucosal C. albicans infections, there is a paucity of information regarding the presence and/or role of T cells in OPC lesions. In pursuit of this, initial chromophore immunohistochemical studies showed a majority of CD8(+) rather than CD4(+) cells equally distributed throughout the buccal mucosa of OPC( ) persons (HIV(-) or HIV(+)), irrespective of blood CD4(+) cell numbers. In contrast, CD8(+) cells in lesions from HIV(+) OPC(+) persons were in significantly higher numbers and concentrated at the lamina propria-epithelium interface, a considerable distance from the Candida at the outer epithelium. Dual fluorescence and confocal microscopy confirmed that the majority of CD8(+), but not CD4(+), cells were T cells by the presence or absence, respectively, of CD3 on each cell type. These results suggest that CD8(+) T cells may be important for oral host defense against OPC, especially when CD4 cell numbers are reduced, with a potential CD8 cell-specific dysfunction associated with susceptibility to OPC. PMID- 12540579 TI - Apoptotic killing and phagocytosis of host cells by the parasite Entamoeba histolytica. AB - The ability of Entamoeba histolytica to kill and phagocytose host cells correlates with parasite virulence. This study addressed the role of apoptotic cell killing and host cell phosphatidylserine exposure in the subsequent phagocytosis of Jurkat T cells by E. histolytica. Ingested host cells were apoptotic, as evidenced by the activation of caspase 3 in 88% +/- 3% (mean and standard deviation [SD] of the mean) of Jurkat cells engulfed by E. histolytica; ingested cells without detectable active caspase 3 were already disrupted and partially digested. That apoptotic cell killing preceded phagocytosis was supported by the demonstration that a higher percentage of amebae ingested apoptotic cells than ingested healthy cells (62% +/- 7% versus 30% +/- 9%, respectively [mean and SD]) (P = 0.008). E. histolytica also ingested apoptotic Jurkat cells more rapidly than necrotic control cells (8.5% +/- 0.4% versus 3.5% +/- 0.7%, respectively [mean and SD]) (P < 0.001). The inhibition of amebic cytotoxicity with D-galactose (which blocks the amebic Gal/GalNAc lectin) blocked the phagocytosis of healthy cells by greater than 80%, providing further evidence that apoptosis preceded engulfment. In contrast, D-galactose blocked the phagocytosis of already apoptotic cells by only 40%, implicating an additional host ligand (besides D-galactose) in amebic engulfment of apoptotic cells. The most characteristic surface change on apoptotic cells is phosphatidylserine exposure. Consistent with a role for host cell phosphatidylserine exposure in amebic ingestion of killed cells, Jurkat cell phosphatidylserine was exposed during incubation with E. histolytica (27% +/- 1% [mean and SD] specific increase at 30 min) (the P value versus the control was 0.0003). Approximately 50% more amebae ingested viable Jurkat cells expressing phosphatidylserine on the outer leaflet of the plasma membrane than ingested control cells (30.3% +/- 2.2% versus 19.8% +/- 1.9%, respectively [mean and SD]) (P = 0.003). By analogy with phagocytic clearance during apoptosis in metazoans, amebic apoptotic host cell killing followed by phagocytosis may limit inflammation and enable amebae to evade the host immune response. PMID- 12540580 TI - Restricted fusion of Chlamydia trachomatis vesicles with endocytic compartments during the initial stages of infection. AB - The chlamydial inclusion occupies a unique niche within the eukaryotic cell that does not interact with endocytic compartments but instead is fusogenic with a subset of sphingomyelin-containing exocytic vesicles. The Chlamydia trachomatis inclusion acquires these distinctive properties by as early as 2 h postinfection as demonstrated by the ability to acquire sphingomyelin, endogenously synthesized from 6[N-[(7-nitrobenzo-2-oxa-1,3-diazol-4-yl)amino]caproylsphingosine] (C(6)-NBD ceramide). The molecular mechanisms involved in transformation of the properties and cellular interactions of the inclusion are unknown except that they require early chlamydial transcription and translation. Although the properties of the inclusion are established by 2 h postinfection, the degree of interaction with endocytic pathways during the brief interval before fusogenicity with an exocytic pathway is established is unknown. Using a combination of confocal and electron microscopy to localize endocytic and lysosomal markers in C. trachomatis infected cells during the early stages of infection, we demonstrate a lack of these markers within the inclusion membrane or lumen of the inclusion to conclude that the nascent chlamydial inclusion is minimally interactive with endosomal compartments during this interval early in infection. Even when prevented from modifying the properties of the inclusion by incubation in the presence of protein synthesis inhibitors, vesicles containing elementary bodies are very slow to acquire lysosomal characteristics. These results imply a two-stage mechanism for chlamydial avoidance of lysosomal fusion: (i) an initial phase of delayed maturation to lysosomes due to an intrinsic property of elementary bodies and (ii) an active modification of the vesicular interactions of the inclusion requiring chlamydial protein synthesis. PMID- 12540581 TI - Restricted expression of shiga toxin binding sites on mucosal epithelium of mouse distal colon. AB - Shiga toxins (Stx) are some of the major virulence factors of enterohemorrhagic Escherichia coli strains such as serotype O157:H7. To explore how Stx might initially gain access to the bloodstream from sites of infection, frozen sections of mouse colon were immunohistochemically examined for binding sites for recombinant binding subunits (Stx1B). Binding sites were selectively expressed on the epithelium in the distal half of the mouse colon, whereas the proximal half did not exhibit any binding sites. In agreement with this observation, we also demonstrated the distal-part-restricted distribution of glycolipids that bind to Stx1B in the mouse colon. For comparison, the binding sites of several control lectins were also examined. Selective binding to the distal part of the colon was not seen with any other control lectins, including Griffonia simplicifolia lectin I isolectin B4 (GS-I-B4), which shares alpha-galactose specificity with Stx1B. Partial overlapping of the specificities of Stx1B and GS-I-B4 was seen by assay with globotriose-conjugated multivalent ligands. The results indicate that Stx1B is stricter in the recognition of carbohydrate determinants than GS-I-B4 when examined with biological ligands. PMID- 12540582 TI - Epithelial cell surface sites involved in the polyvalent adherence of Porphyromonas gingivalis: a convincing role for neuraminic acid and glucuronic acid. AB - We investigated the target structures of the epithelial cells responsible for the attachment of Porphyromonas gingivalis by immunocytofluorimetry, enzyme-linked immunosorbent assay, and confocal microscopy. Integrins (beta1, beta3, and alphaV) and E-cadherin played no significant role. Carbohydrates (such as alpha-D methylglucoside, L-fucose, D- and L-mannose, N-acetylglucosamine, and N acetylgalactosamine) had little inhibitory effect on bacterial binding. Enzymatic treatments of the epithelial membranes and sugar competition studies showed that N-acetylneuraminic acid and glucuronic acid were involved in binding. PMID- 12540583 TI - DNA alkylation damage as a sensor of nitrosative stress in Mycobacterium tuberculosis. AB - One of the cellular consequences of nitrosative stress is alkylation damage to DNA. To assess whether nitrosative stress is registered on the genome of Mycobacterium tuberculosis, mutants lacking an alkylation damage repair and reversal operon were constructed. Although hypersensitive to the genotoxic effects of N-methyl-N'-nitro-N-nitrosoguanidine in vitro, the mutants displayed no phenotype in vivo, suggesting that permeation of nitrosative stress to the level of cytotoxic DNA damage is restricted. PMID- 12540584 TI - Regulation of expression of the Borrelia burgdorferi beta(3)-chain integrin ligand, P66, in ticks and in culture. AB - Borrelia burgdorferi is maintained in an infection cycle between mammalian and arthropod hosts. Appropriate gene expression by B. burgdorferi at different stages of this cycle is probably essential for transmission and establishment of infection. The B. burgdorferi beta(3) integrin ligand P66 is expressed by the bacteria in mammals, laboratory culture, and engorged but not unfed ticks. No in vitro culture conditions in which P66 expression reflected that in the unfed tick were found, suggesting that there are aspects of B. burgdorferi-tick interaction that remain unexplored. PMID- 12540585 TI - Migration through the extracellular matrix by the parasitic protozoan Leishmania is enhanced by surface metalloprotease gp63. AB - Leishmania species engineered to express high levels of the surface metalloprotease gp63 have enhanced capacity of migration through extracellular matrix in vitro. This correlates with gp63 degradation of extracellular matrix components, such as collagen type IV and fibronectin, and suggests an important role for gp63 in the pathogenesis of leishmaniasis. PMID- 12540586 TI - Modulation of J774.1 macrophage L-arginine metabolism by intracellular Mycobacterium bovis BCG. AB - Using a Mycobacterium bovis BCG mutant (AS1) lacking a Bacillus subtilis L arginine transporter homolog, we demonstrate here that the interaction between intracellular mycobacteria and the macrophage with respect to L-arginine transport and metabolism is quite complex. Intracellular AS1 stimulates macrophage L-arginine transport and accumulates 2.5-fold more (3)H label derived from L-arginine than does the wild type. These studies suggest that the accumulation of (3)H label reflects the acquisition of metabolites of L-arginine produced by the macrophage. PMID- 12540587 TI - Influence of extracellular bactericidal agents on bacteria within macrophages. AB - We employed gentamicin-sensitive and -resistant derivatives of Escherichia coli in a macrophage phagocytosis assay that compared lambda bacteriophage and gentamicin as extracellular bactericidal agents. Colony counts and direct microscopic examination of phagocytized E. coli supported the conclusion that gentamicin entered macrophages, even at low concentrations, and contributed to their bactericidal activity. Also, two E. coli strains differing in the ability to express the adhesin of type 1 pili (FimH) were distinguishably different in intracellular survival when lambda was used as the extracellular killing agent but were indistinguishable when gentamicin was employed. PMID- 12540588 TI - Pathogenic potential of environmental Vibrio cholerae strains carrying genetic variants of the toxin-coregulated pilus pathogenicity island. AB - The major virulence factors of toxigenic Vibrio cholerae are cholera toxin (CT), which is encoded by a lysogenic bacteriophage (CTXPhi), and toxin-coregulated pilus (TCP), an essential colonization factor which is also the receptor for CTXPhi. The genes for the biosynthesis of TCP are part of a larger genetic element known as the TCP pathogenicity island. To assess their pathogenic potential, we analyzed environmental strains of V. cholerae carrying genetic variants of the TCP pathogenicity island for colonization of infant mice, susceptibility to CTXPhi, and diarrheagenicity in adult rabbits. Analysis of 14 environmental strains, including 3 strains carrying a new allele of the tcpA gene, 9 strains carrying a new allele of the toxT gene, and 2 strains carrying conventional tcpA and toxT genes, showed that all strains colonized infant mice with various efficiencies in competition with a control El Tor biotype strain of V. cholerae O1. Five of the 14 strains were susceptible to CTXPhi, and these transductants produced CT and caused diarrhea in adult rabbits. These results suggested that the new alleles of the tcpA and toxT genes found in environmental strains of V. cholerae encode biologically active gene products. Detection of functional homologs of the TCP island genes in environmental strains may have implications for understanding the origin and evolution of virulence genes of V. cholerae. PMID- 12540589 TI - UreR, the transcriptional activator of the Proteus mirabilis urease gene cluster, is required for urease activity and virulence in experimental urinary tract infections. AB - Proteus mirabilis, a cause of complicated urinary tract infection, produces urease, an essential virulence factor for this species. UreR, a member of the AraC/XylS family of transcriptional regulators, positively activates expression of the ure gene cluster in the presence of urea. To specifically evaluate the contribution of UreR to urease activity and virulence in the urinary tract, a ureR mutation was introduced into P. mirabilis HI4320 by homologous recombination. The isogenic ureR::aphA mutant, deficient in UreR production, lacked measurable urease activity. Expression was not detected in the UreR deficient strain by Western blotting with monoclonal antibodies raised against UreD. Urease activity and UreD expression were restored by complementation of the mutant strain with ureR expressed from a low-copy-number plasmid. Virulence was assessed by transurethral cochallenge of CBA mice with wild-type and mutant strains. The isogenic ureR::aphA mutant of HI4320 was outcompeted in the urine (P = 0.004), bladder (P = 0.016), and kidneys (P < or = 0.001) 7 days after inoculation. Thus, UreR is required for basal urease activity in the absence of urea, for induction of urease by urea, and for virulence of P. mirabilis in the urinary tract. PMID- 12540590 TI - Insulin activation of phosphatidylinositol 3-kinase in the hypothalamic arcuate nucleus: a key mediator of insulin-induced anorexia. AB - In peripheral tissues, insulin signaling involves activation of the insulin receptor substrate (IRS)-phosphatidylinositol 3-kinase (PI3K) enzyme system. In the hypothalamus, insulin functions with leptin as an afferent adiposity signal important for the regulation of body fat stores and hepatic glucose metabolism. To test the hypothesis that hypothalamic insulin action involves intracellular PI3K signaling, we used histochemical and biochemical methods to determine the effect of insulin on hypothalamic IRS-PI3K activity. Here, we report that insulin induces tyrosine phosphorylation of the insulin receptor and IRS-1 and -2, increases binding of activated IRS-1 and -2 to the regulatory subunit of PI3K, and activates protein kinase B/Akt, a downstream target of PI3K. Using an immunohistochemical technique to detect PI 3,4,5-triphosphate, the main product of PI3K activity, we further demonstrate that in the arcuate nucleus, insulin induced PI3K activity occurs preferentially within cells that contain IRS-2. Finally, we show that the food intake- lowering effects of insulin are reversed by intracerebroventricular infusion of either of two PI3K inhibitors at doses that have no independent feeding effects. These findings support the hypothesis that the IRS-PI3K pathway is a mediator of insulin action in the arcuate nucleus and, combined with recent evidence that leptin activates PI3K signaling in the hypothalamus, provide a plausible mechanism for neuronal cross-talk between insulin and leptin signaling. PMID- 12540591 TI - Is the energy homeostasis system inherently biased toward weight gain? AB - We describe a model of energy homeostasis to better understand neuronal pathways that control energy balance and their regulation by hormonal signals such as insulin and leptin. Catabolic neuronal pathways are those that both reduce food intake and increase energy expenditure (e.g., melanocortin neurons in the hypothalamic arcuate nucleus) and are stimulated by input from insulin and leptin. We propose that in the basal state, catabolic effectors are activated in response to physiological concentrations of leptin and insulin, and that this activation is essential to prevent excessive weight gain. In contrast, anabolic pathways (e.g., neurons containing neuropeptide Y) are those that stimulate food intake and decrease energy expenditure and are strongly inhibited by these same basal concentrations of insulin and leptin. In the basal state, therefore, catabolic effector pathways are activated while anabolic effector pathways are largely inhibited. The response to weight loss includes both activation of anabolic and inhibition of catabolic pathways and is, thus, inherently more vigorous than the response to weight gain (stimulation of already-activated catabolic pathways and inhibition of already-suppressed anabolic pathways). Teleological, molecular, physiological, and clinical aspects of this hypothesis are presented, along with a discussion of currently available supporting evidence. PMID- 12540592 TI - Plasma adiponectin concentrations predict insulin sensitivity of both glucose and lipid metabolism. AB - In animals, the adipocyte-derived hormone adiponectin has been shown to improve insulin sensitivity, a key factor in the pathogenesis of type 2 diabetes. In Pima Indians, high plasma adiponectin levels are associated with increased insulin sensitivity and reduced risk of type 2 diabetes. It is unclear whether this is also the case in white individuals and whether an additional beneficial effect on lipid metabolism exists. We therefore analyzed in nondiabetic individuals the associations between plasma adiponectin concentrations and insulin sensitivity measured by a euglycemic-hyperinsulinemic clamp (n = 262) and estimated by an oral glucose tolerance test (OGTT; n = 636) and serum lipid parameters using correlational analysis. Plasma adiponectin concentrations were positively correlated with insulin sensitivity, both measured with the clamp (r = 0.28, P = 0.0015 in women; r = 0.42, P < 0.0001 in men) and estimated from the OGTT (r = 0.37, P < 0.0001 in women; r = 0.41, P < 0.0001 in men) before and after adjusting for sex and percentage of body fat (all P < 0.001). Fasting triglycerides and the free fatty acid (FFA) concentrations during the OGTT (area under the curve) and at 120 min were negatively correlated in both women and men, whereas HDL was positively correlated with plasma adiponectin concentrations (all P < 0.004). Most notable, these relationships remained significant after adjusting for insulin sensitivity of glucose disposal in addition to sex and percentage of body fat (all P < 0.05). In conclusion, high adiponectin predicts increased insulin sensitivity. This relationship is independent of low body fat mass and affects not only insulin-stimulated glucose disposal but also lipoprotein metabolism and insulin-mediated suppression of postprandial FFA release. This suggests pleiotropic insulin sensitizing effects of adiponectin in humans. PMID- 12540593 TI - Increased intramyocellular lipid concentration identifies impaired glucose metabolism in women with previous gestational diabetes. AB - Women with previous gestational diabetes (pGDM) are frequently insulin-resistant, which could relate to intramyocellular lipid content (IMCL). IMCL were measured with (1)H nuclear magnetic resonance spectroscopy in soleus (IMCL-S) and tibialis anterior muscles (IMCL-T) of 39 pGDM (32 +/- 2 years, waist-to-hip ratio 0.81 +/- 0.01) and 22 women with normal glucose tolerance (NGT; 31 +/- 1 years, 0.76 +/- 0.02) at 4-6 months after delivery. Body fat mass (BFM) was assessed from bioimpedance analysis, insulin sensitivity index (S(I)), and glucose effectiveness (S(G)) from insulin-modified frequently sampled glucose tolerance tests. pGDM exhibited 45% increased BFM, 35% reduced S(I) and S(G) (P < 0.05), and 40% (P < 0.05) and 55% (P < 0.005) higher IMCL-S and IMCL-T, respectively. IMCL related to body fat (BFM P < 0.005, leptin P < 0.03), but only IMCL-T correlated (P < 0.03) with S(I) and glucose tolerance index independent of BMI. Insulin-resistant pGDM (n = 17) had higher IMCL-S (+66%) and IMCL-T (+86%) than NGT and insulin-sensitive pGDM (+28%). IMCL were also higher (P < 0.005, P = 0.05) in insulin-sensitive pGDM requiring insulin treatment during pregnancy and inversely related to the gestational week of GDM diagnosis. Thus, IMCL-T reflects insulin sensitivity, whereas IMCL-S relates to obesity. IMCL could serve as an additional parameter of increased diabetes risk because it identifies insulin resistant pGDM and those who were diagnosed earlier and/or required insulin during pregnancy. PMID- 12540594 TI - Role of leptin in the regulation of glucagon-like peptide-1 secretion. AB - Glucagon-like peptide-1 (GLP-1), released from intestinal endocrine L cells, is a potent insulinotropic hormone. GLP-1 secretion is diminished in obese patients. Because obesity is linked to abnormal leptin signaling, we hypothesized that leptin may modulate GLP-1 secretion. Leptin significantly stimulated GLP-1 secretion (by up to 250% of control) from fetal rat intestinal cells, a mouse L cell line (GLUTag), and a human L cell line (NCI-H716) in a dose-dependent manner (P < 0.05-0.001). The long form of the leptin receptor was shown to be expressed, and leptin induced the phosphorylation of STAT3 in the three cell types. The leptin receptor was also expressed by rodent and human intestinal L cells, and leptin (1 mg/kg i.p.) significantly stimulated GLP-1 secretion in rats and ob/ob mice. To determine the effect of leptin resistance on GLP-1 secretion, C57BL/6 mice were fed a high-fat (45%) or low-fat (10%) diet for 8 weeks. Mice on the high-fat diet became obese; developed glucose intolerance, hyperinsulinemia, and hyperleptinemia; and were leptin resistant. Mice on the high-fat diet also had twofold lower basal plasma GLP-1 and a diminished GLP-1 response to oral glucose, by 28.5 +/- 5.0% (P < 0.05). These results show for the first time that leptin stimulates GLP-1 secretion from rodent and human intestinal L cells, and they suggest that leptin resistance may account for the decreased levels of GLP-1 found in obese humans. PMID- 12540595 TI - Effects of free fatty acids per se on glucose production, gluconeogenesis, and glycogenolysis. AB - Insulin-independent effects of a physiological increase in free fatty acid (FFA) levels on fasting glucose production, gluconeogenesis, and glycogenolysis were assessed by administering [6,6-(2)H(2)]-glucose and deuteriated water ((2)H(2)O) in 12 type 1 diabetic patients, during 6-h infusions of either saline or a lipid emulsion. Insulin was either fully replaced (euglycemic group, n = 6), or underreplaced (hyperglycemic group, n = 6). During saline infusions, plasma FFA levels remained unchanged. Glucose concentrations decreased from 6.7 +/- 0.4 to 5.3 +/- 0.4 mmol/l and 11.9 +/- 1.0 to 10.5 +/- 1.0 mmol/l in the euglycemic and hyperglycemic group, respectively. Accordingly, glucose production declined from 84 +/- 5 to 63 +/- 5 mg x m(-2) x min(-1) and from 84 +/- 5 to 68 +/- 4 mg x m( 2) x min(-1), due to declining rates of glycogenolysis but unaltered rates of gluconeogenesis. During lipid infusions, plasma FFA levels increased twofold. In the euglycemic group, plasma glucose increased from 6.8 +/- 0.3 to 7.8 +/- 0.8 mmol/l. Glucose production declined less in the lipid study than in the saline study due to a stimulation of gluconeogenesis by 6 +/- 1 mg x m(-2) x min(-1) and a decline in glycogenolysis that was 6 +/- 2 mg x m(-2) x min(-1) less in the lipid study than in the saline study. In contrast, in the hyperglycemic group, there were no significant effects of elevated FFA on glucose production, gluconeogenesis, or glycogenolysis. In conclusion, a physiological elevation of plasma FFA levels stimulates glycogenolysis as well as gluconeogenesis and causes mild fasting hyperglycemia. These effects of FFA appear attenuated in the presence of hyperglycemia. PMID- 12540596 TI - Sexual differentiation, pregnancy, calorie restriction, and aging affect the adipocyte-specific secretory protein adiponectin. AB - Adiponectin or adipocyte complement-related protein of 30 kDa (Acrp30) is a circulating protein produced exclusively in adipocytes. Circulating Acrp30 levels have been associated with insulin sensitivity in adult mice and humans, yet the Acrp30 profile over the lifespan and its hormonal regulation in vivo have not been previously described. Hence, we set forth to determine whether hormonal and metabolic changes associated with sexual maturation, reproduction, aging, and calorie restriction affect Acrp30. In mice, Acrp30 levels increase during sexual maturation by 4-fold in males and 10-fold in females. Neonatal castration (CX) allows Acrp30 of adults to reach female levels. CX in adults does not lead to female Acrp30 levels unless glucocorticoid exposure is elevated simultaneously by implant. Ovariectomy of infant mice does not interfere with the pubertal rise of Acrp30. However, ovariectomy in adults increases Acrp30. Estrogen suppressed Acrp30 in mice and 3T3-L1 adipocytes. In parallel to changes in estrogen action, Acrp30 decreased in late gestation but increased in both calorie-restricted and old (anovulatory) mice. The reduction of Acrp30 in lactating dams is consistent with a suppressive effect of prolactin and a stimulating effect of bromocriptine. In summary, Acrp30 levels in serum are under complex hormonal control and may play a key role in determining systemic insulin sensitivity under the respective conditions. PMID- 12540597 TI - Leptin receptor-deficient obese Zucker rats reduce their food intake in response to a systemic supply of calories from glucose. AB - It has been established that leptin exerts a negative control on food intake, allowing one to maintain stable caloric intake over time. The aim of the present study was to investigate whether leptin regulates food intake when a supply of calories is provided by the systemic route. Experiments were carried out in leptin receptor-deficient obese fa/fa rats and lean Fa/fa controls. In both groups, 48 h of glucose infusion reduced food intake in proportion to caloric supply, resulting in virtually no change in total caloric intake as compared to before the infusion. This hypophagic response was reproduced without adding systemic calories, but by increasing glucose and insulin concentrations specifically in the brain through carotid artery infusion. Concomitant intracerebroventricular administration of 5-(tetradecyloxy)-2-furoic acid, an acetyl CoA carboxylase inhibitor that precludes malonyl-CoA synthesis, abolished the restriction of feeding in carotid-infused lean and obese rats. These data indicate that a supply of calories via glucose infusion induces a hypophagic response independent of leptin signaling in the rat, and support the hypothesis that a rise in central malonyl-CoA, triggered by increased glucose and insulin concentrations, participates in this adaptation. This process could contribute to the limiting of hyperphagia, primarily when leptin signaling is altered, as in the obese state. PMID- 12540598 TI - Differential effects of rosiglitazone and metformin on adipose tissue distribution and glucose uptake in type 2 diabetic subjects. AB - We evaluated the effects of rosiglitazone (4 mg b.i.d.) and metformin (1 g b.i.d.) monotherapy for 26 weeks on adipose tissue insulin-stimulated glucose uptake in patients (n = 41) with type 2 diabetes. Before and after the treatment, glucose uptake was measured using 2-[(18)F]fluoro-2-deoxyglucose and positron emission tomography and adipose tissue masses were quantified using magnetic resonance imaging. Rosiglitazone improved insulin-stimulated whole-body glucose uptake by 44% (P < 0.01 vs. placebo). Mean body weight was unchanged in the rosiglitazone group, while it decreased by 2.0 kg in the metformin group (P < 0.05 vs. placebo). In visceral adipose tissue, glucose uptake increased by 29% (from 17.8 +/- 2.0 to 23.0 +/- 2.6 micro mol x kg(-1) x min(-1), P < 0.05 vs. placebo) in the rosiglitazone group but to a lesser extent (17%) in the metformin group (from 16.2 +/- 1.5 to 18.9 +/- 1.7 micro mol x kg(-1) x min(-1), P < 0.05 vs. baseline). Because the visceral adipose tissue mass simultaneously decreased with both treatments (P < 0.05), no change was observed in total visceral glucose uptake per depot. Rosiglitazone significantly enhanced glucose uptake in the femoral subcutaneous area, either when expressed per tissue mass (from 10.8 +/- 1.2 to 17.1 +/- 1.7 micro mol x kg(-1) x min(-1), P < 0.01 vs. placebo) or per whole-fat depot (P < 0.05 vs. placebo). In conclusion, metformin treatment resulted in improvement of glycemic control without enhancement of peripheral insulin sensitivity. The improved insulin sensitivity of the nonabdominal subcutaneous adipose tissue during treatment with rosiglitazone partly explains the enhanced whole-body insulin sensitivity and underlies the central role of adipose tissue for action of peroxisome proliferator-activated receptor gamma agonist in vivo. PMID- 12540599 TI - PPAR-gamma activation mediates adipose depot-specific effects on gene expression and lipoprotein lipase activity: mechanisms for modulation of postprandial lipemia and differential adipose accretion. AB - This study sought to determine whether the adipose depot-specific (subcutaneous [SF] vs. visceral [VF]) action of peroxisome proliferator-activated receptor gamma (PPAR-gamma) agonists on fat deposition extends to the expression of lipoprotein lipase (LPL) and other key adipose lipid metabolism genes, and whether changes in LPL impact triglyceridemia. Rats were fed a standard diet or an obesity-promoting diet for 3 weeks, with or without treatment with COOH, a nonthiazolidinedione PPAR-gamma agonist. Treatment effects were essentially similar in both dietary cohorts. COOH did not affect weight gain, but increased SF (inguinal) fat mass twofold and reduced VF (retroperitoneal) accretion by half. Corresponding depot-specific alterations were observed in mRNA levels of the glucocorticoid-activating enzyme 11beta-hydroxysteroid dehydrogenase 1 (11beta-HSD-1) and the thermogenic modulator uncoupling protein 1 (UCP-1). COOH increased brown adipose tissue (BAT) weight and LPL availability by five- to eightfold. In rats refed standard diet after a 24-h fast, COOH reduced the insulin excursion by half. The agonist increased SF LPL activity and mRNA levels, but had no effect on VF LPL. The two- to threefold postprandial increase in plasma triglycerides (TGs) was abrogated in COOH-treated rats, likely in part because of increased LPL in SF and BAT. Thus PPAR-gamma agonist treatment had a powerful, site-specific effect on adipose metabolism and lipid deposition, and greatly impacted the postprandial handling of TG-rich lipoproteins. These depot specific effects may be mediated by differential regulation of key metabolic genes, including LPL, 11beta-HSD-1, and UCP-1. PMID- 12540600 TI - Role of the fatty acid binding protein mal1 in obesity and insulin resistance. AB - The metabolic syndrome is a cluster of metabolic and inflammatory abnormalities including obesity, insulin resistance, type 2 diabetes, hypertension, dyslipidemia, and atherosclerosis. The fatty acid binding proteins aP2 (fatty acid binding protein [FABP]-4) and mal1 (FABP5) are closely related and both are expressed in adipocytes. Previous studies in aP2-deficient mice have indicated a significant role for aP2 in obesity-related insulin resistance, type 2 diabetes, and atherosclerosis. However, the biological functions of mal1 are not known. Here, we report the generation of mice with targeted null mutations in the mal1 gene as well as transgenic mice overexpressing mal1 from the aP2 promoter/enhancer to address the role of this FABP in metabolic regulation in the presence or absence of obesity. To address the role of the second adipocyte FABP in metabolic regulation in the presence and deficiency of obesity, absence of mal1 resulted in increased systemic insulin sensitivity in two models of obesity and insulin resistance. Adipocytes isolated from mal1-deficient mice also exhibited enhanced insulin-stimulated glucose transport capacity. In contrast, mice expressing high levels of mal1 in adipose tissue display reduced systemic insulin sensitivity. Hence, our results demonstrate that mal1 modulates adipose tissue function and contributes to systemic glucose metabolism and constitutes a potential therapeutic target in insulin resistance. PMID- 12540601 TI - STAT5A promotes adipogenesis in nonprecursor cells and associates with the glucocorticoid receptor during adipocyte differentiation. AB - The differentiation of adipocytes is regulated by the activity of a variety of transcription factors, including peroxidase proliferator-activated receptor (PPAR)-gamma and C/EBPalpha. Our current study demonstrates that ectopic expression of STAT5A, such as that of PPAR-gamma and C/EBPalpha, promotes adipogenesis in two nonprecursor fibroblast cell lines. Using morphologic and biochemical criteria, we have demonstrated that STAT5A and the combination of STAT5A and STAT5B are sufficient to induce the expression of early and late adipogenic markers in BALB/c and NIH-3T3 cells. Yet, the ectopic expression of STAT5B alone does not induce the expression of adipocyte genes, but enhances the induction of these genes in cells also expressing STAT5A. This finding suggests that STAT5A and STAT5B do not function identically in adipocytes. In addition, these studies demonstrate that the phosphorylation of STAT5 proteins may play a role in adipogenesis. Moreover, we have shown that STAT5A is associated with the glucocorticoid receptor during adipogenesis in a highly regulated manner. PMID- 12540602 TI - Hyperactivity and reduced energy cost of physical activity in serotonin 5-HT(2C) receptor mutant mice. AB - We have observed late-onset obesity in mutant mice lacking the serotonin 5-HT(2C) receptor. Despite chronically elevated food intake, young adult mutants exhibit neither elevated adiposity nor altered glucose or fat homeostasis. However, obesity subsequently develops after 6 months of age without increases in their level of hyperphagia. In this study, we investigated determinants of energy expenditure in 5-HT(2C) receptor mutant mice. Young adult mutants displayed patterns of elevated activity levels that were enhanced by fasting and tightly associated with repeated visits to a food source. Surprisingly, subsequent obesity development occurred despite persisting locomotor hyperactivity and without age-related declines in resting metabolic rate. Rather, substantial reductions in the energy cost of locomotor activity (LA) were observed in 5 HT(2C) receptor mutant mice. Moreover, both mutant and wild-type mice displayed age-related declines in the energy cost of LA, indicating that this process may be regulated by both aging and serotonergic signaling. These results indicate that a mutation of the 5-HT(2C) receptor gene (htr2c) increases LA, which contributes to the maintenance of normal body composition in young adult mutants despite their hyperphagia. Moreover, age-dependent reductions in the energy cost of physical activity could contribute to the subsequent development of late-onset obesity in 5-HT(2C) receptor mutant mice. PMID- 12540603 TI - NOD congenic mice genetically protected from autoimmune diabetes remain resistant to transplantation tolerance induction. AB - The loss of self-tolerance leading to autoimmune type 1 diabetes in the NOD mouse model involves at least 19 genetic loci. In addition to their genetic defects in self-tolerance, NOD mice resist peripheral transplantation tolerance induced by costimulation blockade using donor-specific transfusion and anti-CD154 antibody. Hypothesizing that these two abnormalities might be related, we investigated whether they could be uncoupled through a genetic approach. Diabetes-resistant NOD and C57BL/6 stocks congenic for various reciprocally introduced Idd loci were assessed for their ability to be tolerized. Surprisingly, in NOD congenic mice that are almost completely protected from diabetes, costimulation blockade failed to prolong skin allograft survival. In reciprocal C57BL/6 congenic mice with NOD derived Idd loci, skin allograft survival was readily prolonged by costimulation blockade. These data indicate that single or multiple combinations of evaluated Idd loci that dramatically reduce diabetes frequency do not correct resistance to peripheral transplantation tolerance induced by costimulation blockade. We suggest that mechanisms controlling autoimmunity and transplantation tolerance in NOD mice are not completely overlapping and are potentially distinct, or that the genetic threshold for normalizing the transplantation tolerance defect is higher than that for preventing autoimmune diabetes. PMID- 12540604 TI - T-cell compartments of prediabetic NOD mice. AB - Given the importance of the NOD mouse as a model of type 1 diabetes, there is a surprising lack of published information on the overall composition of the thymic and peripheral T-cell compartments. In this study, we revisited some earlier reports of T-cell abnormalities in this strain and examined a number of additional parameters to provide a global view of T-cells in prediabetic NOD mice. In some cases, we concur with past conclusions, but in other important areas, we find that NOD mice closely resemble nonautoimmune strains. Specifically, and contrary to published reports, the thymocyte subset distribution, the rate and composition of thymic export, and the composition of the peripheral T-cell pool, including the proportion of CD25(+)CD4(+) T-cells, are essentially normal in prediabetic NOD mice. These factors are therefore unlikely to be involved in the loss of tolerance that leads to autoimmunity within this strain. PMID- 12540605 TI - Acute shock induced by antigen vaccination in NOD mice. AB - Type 1 diabetes in NOD mice can be prevented through autoantigen vaccination by shifting lymphocyte differentiation toward a T-helper 2 (Th(2)) response. However, in other models of autoimmunity, this approach may be accompanied by unexpected triggering of Th(2)-dependent anaphylactic shock. To test the safety of vaccination therapy in the NOD mouse model, we evaluated the effects of immunization with a wide battery of antigens in NOD, BALB/c, and C57BL/6 mice. Surprisingly, a nondiabetogenic antigen, hen egg white lysozyme, induced severe shock exclusively in NOD mice (shock in 11 of 11 mice, lethal in 3 mice). Shock severity was further increased by a more pronounced Th(2) setting generated by 1alpha,25(OH)(2)D(3) administration (17 of 17 mice, lethal in 14 mice, P < 0.0001). Pretreatment with dexamethasone resulted in full rescue, indicating an immune-mediated mechanism. Serum IgE levels and Th(1)/Th(2) cytokine profile analysis showed that the shock phenomenon was paralleled by a Th(2) response. mRNA expression of platelet-activating factor receptor (PAF-R) was significantly higher in NOD mice (P < 0.01) and was further increased by 1alpha,25(OH)(2)D(3). Pretreatment with WEB2086 (PAF-R antagonist) again protected all mice from lethal shock, indicating PAF as an anaphylaxis effector. In conclusion, in NOD mice, vaccination leading to a Th(2) immune shift can result in a lethal anaphylactic reaction. PMID- 12540606 TI - Accelerated diabetes in rat insulin promoter-tumor necrosis factor-alpha transgenic nonobese diabetic mice lacking major histocompatibility class II molecules. AB - The major predisposing genetic component in type 1 diabetes maps to the major histocompatibility complex locus in both mice and humans. To verify the HLA class II association with disease pathogenesis, we adopted the transgenic approach. Expression of HLA-DQ8, the molecule showing the strongest association with human type 1 diabetes, in the diabetes-predisposing milieu of NOD mice in the absence of the endogenous class II molecule I-A(g7) did not render susceptibility to type 1 diabetes. To study if providing a local proinflammatory environment would lead to diabetes in these mice, Abeta(o).NOD.DQ8 were bred with C57BL/6 mice expressing tumor necrosis factor (TNF)-alpha in the beta-cells of the islets of Langerhans. Surprisingly, although diabetes was evident in the F1 intercross expressing rat insulin promoter (RIP)-TNF, offspring lacking either endogenous or transgenic class II molecules developed accelerated diabetes with high frequency in both sexes. Moreover, expression of any functional class II molecule seemed to confer significant protection from diabetes in this model. Thus, neonatal expression of TNF-alpha in an islet-specific manner bypassed the requirement of CD4(+) T-cells and resulted in diabetes that could be mediated by CD8(+) T-cells. We also show for the first time that diabetes in NOD.RIP-TNF mice can occur independent of inheritance of NOD-derived idd1. PMID- 12540607 TI - Molecular regulation of monocyte chemoattractant protein-1 expression in pancreatic beta-cells. AB - Pancreatic beta-cells are selectively destroyed during the course of type 1 diabetes. In the early stages of the disease, inflammatory infiltrates of mononuclear cells, containing predominantly monocytes and T-cells, are present in the islets (insulitis). Chemokines, such as monocyte chemoattractant protein-1 (MCP-1), play a key role in the recruitment and activation of these immunocytes. We have previously described cytokine-induced MCP-1 gene expression in human and rat pancreatic islets. In the present study, the transcriptional regulation by cytokines of the rat MCP-1 gene in fluorescence-activated cell sorting-purified rat beta-cells, insulin-producing INS-1E cells, and RINm5F cells was investigated. Transient transfections with luciferase-reporter constructs identified an interleukin (IL)-1beta-responsive enhancer region between -2,180 bp and -2,478 bp. Mutation of either of the two nuclear factor (NF)-kappaB sites present in this region abrogated IL-1beta-induced MCP-1 promoter activity. Binding of NF-kappaB to the two sites was shown in vitro by gel shift assays, while supershift assays revealed the presence of p65/p50 heterodimers and p65 homodimers. In vivo binding of NF-kappaB was confirmed by chromatin immunoprecipitation assay. Blocking of NF-kappaB activation in cytokine-exposed primary beta-cells by an adenovirus overexpressing a nondegradable form of IkappaBalpha or by pyrrolidine dithiocarbamate decreased IL-1beta-induced MCP-1 mRNA expression. We conclude that NF-kappaB plays an important role for MCP-1 expression in beta-cells. This transcription factor may be an interesting target for ex vivo gene therapy before islet transplantation. PMID- 12540608 TI - Stimulation of insulin secretion by denatonium, one of the most bitter-tasting substances known. AB - Denatonium, one of the most bitter-tasting substances known, stimulated insulin secretion in clonal HIT-T15 beta-cells and rat pancreatic islets. Stimulation of release began promptly after exposure of the beta-cells to denatonium, reached peak rates after 4-5 min, and then declined to near basal values after 20-30 min. In islets, no effect was observed at 2.8 mmol/;l glucose, whereas a marked stimulation was observed at 8.3 mmol/;l glucose. No stimulation occurred in the absence of extracellular Ca(2+) or in the presence of the Ca(2+)-channel blocker nitrendipine. Stimulated release was inhibited by alpha(2)-adrenergic agonists. Denatonium had no direct effect on voltage-gated calcium channels or on cyclic AMP levels. There was no evidence for the activation of gustducin or transducin in the beta-cell. The results indicate that denatonium stimulates insulin secretion by decreasing KATP channel activity, depolarizing the beta-cell, and increasing Ca(2+) influx. Denatonium did not displace glybenclamide from its binding sites on the sulfonylurea receptor (SUR). Strikingly, it increased glybenclamide binding by decreasing the K(d). It is concluded that denatonium, which interacts with K(+) channels in taste cells, most likely binds to and blocks Kir6.2. A consequence of this is a conformational change in SUR to increase the SUR/glybenclamide binding affinity. PMID- 12540609 TI - Role of endogenous glucagon-like peptide-1 in islet regeneration after partial pancreatectomy. AB - A reduction in beta-cell mass is an important causative factor in type 1 and type 2 diabetes. Glucagon-like peptide-1 (GLP-1) and the long-acting agonist exendin 4 (Ex-4) expand beta-cell mass by stimulating neogenesis and proliferation. In the partial pancreatectomy (Ppx) model, exogenous Ex-4 promotes islet regeneration, leading to sustained improvement in glucose tolerance. In this study, we investigate the potential role of endogenous GLP-1 in islet growth. We examined beta-cell mass regeneration after 70% Ppx in mice receiving the GLP-1 antagonist Ex9-39 and in GLP-1R(-/-) mice. In Ex9-39-treated sham-operated mice, persistent fasting hyperglycemia was observed, but beta-cell mass was not diminished. In pancreatectomized mice, persistent glucose intolerance was noted, but this was not further exacerbated by Ex9-39. Accordingly, beta-cell mass recovery of Ppx mice was not impaired by Ex9-39. In contrast, GLP-1R(-/-) CD1 mice showed worse glucose intolerance after Ppx compared with wild-type CD1 Ppx mice, and this correlated with a significant defect in beta-cell mass regeneration. The recovery of beta-cell mass differed markedly in the BALB/c and CD1 control mice, indicating a significant role of genetic background in the regulation of beta cell mass. These studies point to a role for endogenous GLP-1 in beta-cell regeneration after Ppx in mice. PMID- 12540610 TI - Increased dietary fat promotes islet amyloid formation and beta-cell secretory dysfunction in a transgenic mouse model of islet amyloid. AB - Transgenic mice expressing the amyloidogenic human islet amyloid polypeptide (hIAPP) in their islet beta-cells are a model of islet amyloid formation as it occurs in type 2 diabetes. Our hIAPP transgenic mice developed islet amyloid when fed a breeder chow but not regular chow. Because the breeder chow contained increased amounts of fat, we hypothesized that increased dietary fat enhances islet amyloid formation. To test this hypothesis, we fed male hIAPP transgenic and nontransgenic control mice diets containing 15% (low fat), 30% (medium fat), or 45% (high fat) of calories derived from fat for 12 months, and we measured islet amyloid, islet endocrine cell composition, and beta-cell function. Increased dietary fat in hIAPP transgenic mice was associated with a dose dependent increase in both the prevalence (percentage of islets containing amyloid deposits; 34 +/- 8, 45 +/- 8, and 58 +/- 10%, P < 0.05) and severity (percentage of islet area occupied by amyloid; 0.8 +/- 0.5, 1.0 +/- 0.5, and 4.6 +/- 2.5%, P = 0.05) of islet amyloid. In addition, in these hIAPP transgenic mice, there was a dose-dependent decrease in the proportion of islet area comprising beta-cells, with no significant change in islet size. In contrast, nontransgenic mice adapted to diet-induced obesity by increasing their islet size more than twofold. Increased dietary fat was associated with impaired insulin secretion in hIAPP transgenic (P = 0.05) but not nontransgenic mice. In summary, dietary fat enhances both the prevalence and severity of islet amyloid and leads to beta-cell loss and impaired insulin secretion. Because both morphologic and functional defects are present in hIAPP transgenic mice, this would suggest that the effect of dietary fat to enhance islet amyloid formation might play a role in the pathogenesis of the islet lesion of type 2 diabetes in humans. PMID- 12540611 TI - The influence of GLP-1 on glucose-stimulated insulin secretion: effects on beta cell sensitivity in type 2 and nondiabetic subjects. AB - The intestinally derived hormone glucagon-like peptide 1 (GLP-1) (7-36 amide) has potent effects on glucose-mediated insulin secretion, insulin gene expression, and beta-cell growth and differentiation. It is, therefore, considered a potential therapeutic agent for the treatment of type 2 diabetes. However, the dose-response relationship between GLP-1 and basal and glucose-stimulated prehepatic insulin secretion rate (ISR) is currently not known. Seven patients with type 2 diabetes and seven matched nondiabetic control subjects were studied. ISR was determined during a graded glucose infusion of 2, 4, 6, 8, and 12 mg x kg(-1) x min(-1) over 150 min on four occasions with infusion of saline or GLP-1 at 0.5, 1.0, and 2.0 pmol x kg(-1) x min(-1). GLP-1 enhanced ISR in a dose dependent manner during the graded glucose infusion from 332 +/- 51 to 975 +/- 198 pmol/kg in the patients with type 2 diabetes and from 711 +/- 123 to 2,415 +/ 243 pmol/kg in the control subjects. The beta-cell responsiveness to glucose, expressed as the slope of the linear relation between ISR and the glucose concentration, increased in proportion to the GLP-1 dose to 6 times relative to saline at the highest GLP-1 dose in the patients and 11 times in the control subjects, but it was 3 to 5 times lower in the patients with type 2 diabetes compared with healthy subjects at the same GLP-1 dose. During infusion of GLP-1 at 0.5 pmol x kg(-1) x min(-1) in the patients, the slope of ISR versus glucose became indistinguishable from that of the control subjects without GLP-1. Our results show that GLP-1 increases insulin secretion in patients with type 2 diabetes and control subjects in a dose-dependent manner and that the beta-cell responsiveness to glucose may be increased to normal levels with a low dose of GLP-1 infusion. Nevertheless, the results also indicate that the dose-response relation between beta-cell responsiveness to glucose and GLP-1 is severely impaired in patients with type 2 diabetes. PMID- 12540612 TI - Gene transfer of manganese superoxide dismutase extends islet graft function in a mouse model of autoimmune diabetes. AB - Islet transplantation is a promising cure for diabetes. However, inflammation, allorejection, and recurrent autoimmune damage all may contribute to early graft loss. Pancreatic islets express lower levels of antioxidant genes than most other tissues of the body, and beta-cells in particular are sensitive to oxidative damage. Therefore, damage from oxidative stress may pose a major obstacle to islet replacement therapy in that both the islet isolation and transplantation processes generate oxygen radicals. To determine whether antioxidant gene overexpression in isolated pancreatic islets can prevent oxidative damage and prolong islet function after transplantation, we used the NOD mouse model to study oxidative stress encountered during both transplantation and autoimmune attack. We transferred an antioxidant gene, manganese superoxide dismutase (MnSOD), by adenoviral infection into isolated islets that were transplanted into streptozotocin-treated NODscid recipient mice. Functioning islet grafts were subsequently exposed to diabetogenic spleen cells and monitored until graft failure. The results show that islet grafts overexpressing MnSOD functioned approximately 50% longer than control grafts. This significant prolongation of graft function suggests that the antioxidant activity of MnSOD is beneficial to transplanted islet survival and may be used in combination with other strategies aimed at islet graft protection. PMID- 12540613 TI - Differential effects of glucose and glyburide on energetics and Na+ levels of betaHC9 cells: nuclear magnetic resonance spectroscopy and respirometry studies. AB - In the present study, noninvasive (31)P and (23)Na(+)-nuclear magnetic resonance (NMR) technology and respirometry were used to compare the effect of high glucose (30 mmol/l) with the effect of the antidiabetic sulfonylurea (SU) compound glyburide (GLY) on energy metabolism, Na(+) flux, insulin, and cAMP release of continuously superfused beta-HC9 cells encapsulated in microscopic agarose beads. Both high glucose and GLY increased oxygen consumption in beta-HC9 cells (15-30%) with a maximal effect at 8 mmol/l for glucose and at 250 nmol/l for GLY. At the same time, insulin release from beta-cells increased by 15- and 25-fold with high glucose or GLY, respectively. The P-creatine (PCr) level was greatly increased and inorganic phosphate (P(i)) was decreased with 30 mmol/l glucose in contrast to the decreased level of PCr and increased P(i) with GLY. ATP levels remained unchanged during both interventions. Studies on isolated mitochondria of beta-HC9 cells showed that GLY added to mitochondria oxidizing glutamine or glutamate abolished the stimulation of respiration by ADP (state 3) meanwhile leaving state 3 respiration unchanged during oxidation of other substrates. Exposure of beta HC9 cells to 5 mmol/l glucose decreased intracellular Na(+) levels monitored by (23)Na(+)-NMR spectroscopy and 30 mmol/l glucose resulted in a further decrease in cytosolic Na(+). In contrast, Na(+) increased when 1 micro mol/l GLY was added to the perfusate containing 5 mmol/l glucose. These data support the hypothesis that glucose activates the beta-cell through a "push mechanism" due to substrate pressure enhancing fuel flux, energy production, and extrusion of Na(+) from the cells in contrast to SU receptor (SUR)-1 inhibitors, which may modify intermediary and energy metabolism secondarily through a "pull mechanism" due to higher energy demand resulting from increased ion fluxes and the exocytotic work load. PMID- 12540614 TI - BETA2 activates transcription from the upstream glucokinase gene promoter in islet beta-cells and gut endocrine cells. AB - Glucokinase (GK) gene transcription initiates in the islet (beta-cell), gut, and brain from promoter sequences residing approximately 35 kbp upstream from those used in liver. Expression of betaGK is controlled in beta-cells by cell-enriched (i.e. pancreatic duodenal homeobox 1 [PDX-1]) and ubiquitously (i.e., Pal) distributed factors that bind to and activate from conserved sequence motifs within the upstream promoter region (termed betaGK). Here, we show that a conserved E-box element also contributes to control in the islet and gut. betaGK promoter-driven reporter gene activity was diminished by mutating the specific sequences involved in E-box-mediated basic helix-loop-helix factor activator binding in islet beta-cells and enteroendocrine cells. Gel shift assays demonstrated that the betaGK and insulin gene E-box elements formed the same cell enriched (BETA2:E47) and generally distributed (upstream stimulatory factor [USF]) protein-DNA complexes. betaGK E-box-driven activity was stimulated in cotransfection assays performed in baby hamster kidney (BHK) cells with BETA2 and E47, but not USF. Chromatin immunoprecipitation assays performed with BETA2 antisera showed that BETA2 occupies the upstream promoter region of the endogenous betaGK gene in beta-cells. We propose that BETA2 (also termed NeuroD1) regulates betaGK promoter activity. PMID- 12540615 TI - Dominant negative pathogenesis by mutant proinsulin in the Akita diabetic mouse. AB - Autosomal dominant diabetes in the Akita mouse is caused by mutation of the insulin 2 gene, whose product replaces a cysteine residue that is engaged in the formation of an intramolecular disulfide bond. These heterozygous mice exhibit severe insulin deficiency despite coexpression of normal insulin molecules derived from three other wild-type alleles of the insulin 1 and 2 genes. Although the results of our previous study suggested that the mutant proinsulin 2 is misfolded and blocked in the transport from the endoplasmic reticulum to the Golgi apparatus, its dominant negative nature has not been fully characterized. In the present study, we investigated the possible pathogenic mechanisms induced by the mutant proinsulin 2. There is no evidence that the mutant proinsulin 2 attenuates the overall protein synthesis rate or promotes the formation of aberrant disulfide bonds. The trafficking of constitutively secreted alkaline phosphatase, however, is significantly decreased in the islets of Akita mice, indicating that the function of early secretory pathways is nonspecifically impaired. Morphological analysis has revealed that secretory pathway organelle architecture is progressively devastated in the beta-cells of Akita mice. These findings suggest that the organelle dysfunction resulting from the intracellular accumulation of misfolded proinsulin 2 is primarily responsible for the defect of coexisting wild-type insulin secretion in Akita beta-cells. PMID- 12540616 TI - Connexin 36 controls synchronization of Ca2+ oscillations and insulin secretion in MIN6 cells. AB - Cx36 is the predominant connexin isoform expressed by pancreatic beta-cells. However, little is known about the role of this protein in the functioning of insulin-secreting cells. To address this question, we searched for a cell line expressing Cx36 and having glucose-induced insulin secretion comparable to that of primary beta-cells. By evaluating Cx36 expression in MIN6, betaTC3, RIN2A, INS1, and HIT cell lines, which differ in their sensitivity to glucose, we found that wild-type MIN6 cells fit these requirements. Therefore, we stably transfected MIN6 cells with a cDNA coding for a Cx36 antisense sequence to study the role of Cx36 in these cells. Independent clones of MIN6 cells were obtained that had a markedly reduced Cx36 expression. Loss of Cx36 decreased functional gap junctional conductance in these clones. This alteration impaired the synchronization of glucose-induced [Ca(2+)](i) oscillations and insulin secretion in response to glucose, to secretagogues that increase [cAMP](i), and to depolarizing conditions. These data provide the first evidence that Cx36-made channels 1) mediate functional coupling in MIN6 cells, 2) provide for synchronous [Ca(2+)](i) oscillations, and 3) are necessary for proper insulin secretion in response to metabolizable and nonmetabolizable secretagogues. PMID- 12540617 TI - Pax-6 activates endogenous proglucagon gene expression in the rodent gastrointestinal epithelium. AB - The proglucagon gene encodes pancreatic glucagon and the glucagon-like peptides, which exert diverse effects on nutrient absorption and assimilation. The therapeutic potential of glucagon-like peptide-1 (GLP-1) has fostered interest in development of cellular engineering approaches to augment endogenous intestinal derived GLP-1 for the treatment of type 2 diabetes. We have used adenovirus technology to examine the potential roles of the transcription factors Cdx-2/3 and Pax-6 as activators of endogenous proglucagon gene expression in enteroendocrine cell lines and in nontransformed rat intestinal cells. Adenoviral expressed Cdx-2/3 and Pax-6 activated proglucagon promoter-luciferase activity in baby hamster kidney (BHK) fibroblasts, HEK 293 cells, and enteroendocrine cell lines. Pax-6, but not Cdx-2/3, induced expression of the endogenous proglucagon gene in enteroendocrine cell lines, but not in heterologous fibroblasts. Furthermore, transduction of primary rat intestinal cell cultures in vitro, or the rat colonic epithelium in vivo, with Ad-Pax-6 activated endogenous proglucagon gene expression. These data demonstrate that Pax-6, but not Cdx-2/3, is capable of activating the endogenous proglucagon gene in both immortalized enteroendocrine cells and the nontransformed intestinal epithelium in vivo. PMID- 12540618 TI - Age-dependent changes in metabolism, contractile function, and ischemic sensitivity in hearts from db/db mice. AB - Glucose and palmitate metabolism and contractile function were measured with ex vivo perfused working hearts from control (db/+) and diabetic (db/db) female mice at 6, 10-12, and 16-18 weeks of age. Palmitate oxidation was increased by 2.2 fold in 6-week-old db/db hearts and remained elevated in 10- to 12- and 16- to 18 week-old hearts. Carbohydrate oxidation was normal at 6 weeks but was reduced to 27 and 23% of control at 10-12 and 16-18 weeks, respectively. At 6 weeks, db/db hearts exhibited a slight reduction in mechanical function, whereas marked signs of dysfunction were evident at 10-12 and 16-18 weeks. Mechanical function after ischemia-reperfusion was examined in hearts from male mice; at 6 weeks, db/db hearts showed normal recovery, whereas at 12 weeks it was markedly reduced. Fatty acid oxidation was the predominant substrate used after reperfusion. Thus, diabetic db/db hearts exhibit signs of a progressive cardiomyopathy; increased fatty acid oxidation preceded reductions in carbohydrate oxidation. Postischemic recovery of function was reduced in db/db hearts, in parallel with age-dependent changes in normoxic contractile performance. Finally, peroxisome proliferator activated receptor-alpha treatment (3 weeks) did not affect sensitivity to ischemia-reperfusion, even though carbohydrate oxidation was increased and palmitate oxidation was decreased. PMID- 12540619 TI - Inflammation-sensitive plasma proteins, diabetes, and mortality and incidence of myocardial infarction and stroke: a population-based study. AB - This study explores the relationship of inflammation-sensitive plasma proteins (ISPs) with the prevalence of diabetes and the interrelationships between ISPs and diabetes in the prediction of death and incidence of myocardial infarction and stroke. Plasma levels of fibrinogen, alpha1-antitrypsin, haptoglobin, ceruloplasmin, and orosomucoid were assessed in 6,050 men, aged 28-61 years. All cause and cardiovascular mortality and incidence of myocardial infarction and stroke were monitored over 18.7 +/- 3.7 years. Prevalence of diabetes (n = 321) was significantly associated with ISP levels among overweight and obese men but not among men with BMI <25 kg/m(2). The association was similar for insulin resistance according to homeostasis model assessment. High ISP levels (two or more ISPs in the top quartile) increased the cardiovascular risk among diabetic men. The risk factor-adjusted relative risks for cardiovascular mortality, myocardial infarction, and stroke were 2.8 (CI 1.8-4.5), 2.2 (1.5-3.2), and 2.5 (1.4-4.6), respectively, for diabetic men with high ISP levels (reference: nondiabetic men with low ISP levels). The corresponding risks for diabetic men with low ISP levels were 1.8 (1.1-3.0), 1.3 (0.8-2.1), and 1.2 (0.6-2.5), respectively. In conclusion, in this population-based cohort, diabetes was associated with increased ISP levels among overweight and obese men but not among men with normal weight. High ISP levels increased the cardiovascular risk similarly in diabetic as compared with nondiabetic men. PMID- 12540620 TI - Preferential stiffening of central over peripheral arteries in type 2 diabetes. AB - Arterial stiffness affects cardiac functions, peripheral circulation, and cardiovascular mortality. We examined whether arterial stiffness in different regions is equally affected by diabetes and other factors. The subjects were 161 patients with type 2 diabetes and 129 healthy subjects comparable in age and sex. Arterial stiffness was evaluated by measuring pulse wave velocity (PWV) in the heart-carotid, heart-brachial, heart-femoral, and femoral-ankle segments using an automatic device. The diabetic patients had greater PWV than the healthy subjects in the four arterial regions, and the effect of diabetes on PWV was greater in the heart-carotid and heart-femoral segments (central) than in the heart-brachial and femoral-ankle regions (peripheral). PWV increased with age in the four arterial regions, and the effect of age on PWV was greater in the central than in peripheral arteries. In multiple regression analysis, age and systolic blood pressure had significant impacts on PWV of the four regions, whereas diabetes was significantly associated only with PWV of the central arteries. In contrast, sex was associated with PWV of the peripheral arteries. Thus, type 2 diabetes had greater impact on PWV of the central arteries, and different factors were involved in PWV among different arterial regions. PMID- 12540621 TI - Effects of insulin resistance and type 2 diabetes on lipoprotein subclass particle size and concentration determined by nuclear magnetic resonance. AB - The insulin resistance syndrome (IRS) is associated with dyslipidemia and increased cardiovascular disease risk. A novel method for detailed analyses of lipoprotein subclass sizes and particle concentrations that uses nuclear magnetic resonance (NMR) of whole sera has become available. To define the effects of insulin resistance, we measured dyslipidemia using both NMR lipoprotein subclass analysis and conventional lipid panel, and insulin sensitivity as the maximal glucose disposal rate (GDR) during hyperinsulinemic clamps in 56 insulin sensitive (IS; mean +/- SD: GDR 15.8 +/- 2.0 mg. kg(-1). min(-1), fasting blood glucose [FBG] 4.7 +/- 0.3 mmol/l, BMI 26 +/- 5), 46 insulin resistant (IR; GDR 10.2 +/- 1.9, FBG 4.9 +/- 0.5, BMI 29 +/- 5), and 46 untreated subjects with type 2 diabetes (GDR 7.4 +/- 2.8, FBG 10.8 +/- 3.7, BMI 30 +/- 5). In the group as a whole, regression analyses with GDR showed that progressive insulin resistance was associated with an increase in VLDL size (r = -0.40) and an increase in large VLDL particle concentrations (r = -0.42), a decrease in LDL size (r = 0.42) as a result of a marked increase in small LDL particles (r = -0.34) and reduced large LDL (r = 0.34), an overall increase in the number of LDL particles (r = -0.44), and a decrease in HDL size (r = 0.41) as a result of depletion of large HDL particles (r = 0.38) and a modest increase in small HDL (r = -0.21; all P < 0.01). These correlations were also evident when only normoglycemic individuals were included in the analyses (i.e., IS + IR but no diabetes), and persisted in multiple regression analyses adjusting for age, BMI, sex, and race. Discontinuous analyses were also performed. When compared with IS, the IR and diabetes subgroups exhibited a two- to threefold increase in large VLDL particle concentrations (no change in medium or small VLDL), which produced an increase in serum triglycerides; a decrease in LDL size as a result of an increase in small and a reduction in large LDL subclasses, plus an increase in overall LDL particle concentration, which together led to no difference (IS versus IR) or a minimal difference (IS versus diabetes) in LDL cholesterol; and a decrease in large cardioprotective HDL combined with an increase in the small HDL subclass such that there was no net significant difference in HDL cholesterol. We conclude that 1) insulin resistance had profound effects on lipoprotein size and subclass particle concentrations for VLDL, LDL, and HDL when measured by NMR; 2) in type 2 diabetes, the lipoprotein subclass alterations are moderately exacerbated but can be attributed primarily to the underlying insulin resistance; and 3) these insulin resistance-induced changes in the NMR lipoprotein subclass profile predictably increase risk of cardiovascular disease but were not fully apparent in the conventional lipid panel. It will be important to study whether NMR lipoprotein subclass parameters can be used to manage risk more effectively and prevent cardiovascular disease in patients with the IRS. PMID- 12540622 TI - Prediction of type 2 diabetes using simple measures of insulin resistance: combined results from the San Antonio Heart Study, the Mexico City Diabetes Study, and the Insulin Resistance Atherosclerosis Study. AB - To determine and formally compare the ability of simple indexes of insulin resistance (IR) to predict type 2 diabetes, we used combined prospective data from the San Antonio Heart Study, the Mexico City Diabetes Study, and the Insulin Resistance Atherosclerosis Study, which include well-characterized cohorts of non Hispanic white, African-American, Hispanic American, and Mexican subjects with 5 8 years of follow-up. Poisson regression was used to assess the ability of each candidate index to predict incident diabetes at the follow-up examination (343 of 3,574 subjects developed diabetes). The areas under the receiver operator characteristic (AROC) curves for each index were calculated and statistically compared. In pooled analysis, Gutt et al.'s insulin sensitivity index at 0 and 120 min (ISI(0,120)) displayed the largest AROC (78.5%). This index was significantly more predictive (P < 0.0001) than a large group of indexes (including those by Belfiore, Avignon, Katz, and Stumvoll) that had AROC curves between 66 and 74%. These findings were essentially similar both after adjustment for covariates and when analyses were conducted separately by glucose tolerance status and ethnicity/study subgroups. In conclusion, we found substantial differences between published IR indexes in the prediction of diabetes, with ISI(0,120) consistently showing the strongest prediction. This index may reflect other aspects of diabetes pathogenesis in addition to IR, which might explain its strong predictive abilities despite its moderate correlation with direct measures of IR. PMID- 12540623 TI - Altered homeostatic adaptation of first- and second-phase beta-cell secretion in the offspring of patients with type 2 diabetes: studies with a minimal model to assess beta-cell function. AB - We adapted a minimal model to assess beta-cell function during a hyperglycemic glucose clamp and to uncover peculiar aspects of the relationship among beta-cell function, plasma glucose, and insulin sensitivity (IS) in offspring of Caucasian patients with type 2 diabetes (OfT2D). We pooled two data sets of OfT2D (n = 69) and control subjects (n = 45) with normal glucose regulation. Plasma C-peptide was measured during a hyperglycemic clamp ( approximately 10 mmol/l) to quantify model-based first-phase secretion and glucose sensitivity of second-phase secretion (beta). IS was quantified during the hyperglycemic clamp. In the pooled data, first-phase secretion was linearly and negatively related to fasting plasma glucose, but not IS; OfT2D lay on a distinct line shifted to the left of the control subjects. In contrast, beta was negatively related to IS, and OfT2D lay on a distinct line shifted more and more to the left of the control subjects, as IS was worse. Thus, in OfT2D lower beta-cell adaptive responses exist between ambient glycemia and first-phase insulin secretion and between IS and second phase secretion. Under conditions leading to decreased insulin sensitivity, these disturbed relationships may lead to progression to diabetes in OfT2D. PMID- 12540624 TI - Diabetes activates cell death pathway after transient focal cerebral ischemia. AB - It is well known that diabetes aggravates brain damage in experimental and clinical stroke subjects. Diabetes accelerates maturation of neuronal damage, increases infarct volume, and induces postischemic seizures. The mechanism by which diabetes increases ischemic brain damage is still elusive. Our previous experiments indicate that mitochondria dysfunction may play a role in neuronal death. The objective of this study is to determine whether streptozotocin-induced diabetes activates cell death pathway after a brief period of focal cerebral ischemia. Both diabetic and nondiabetic rats were subjected to 30 min of transient middle cerebral artery occlusion, followed by 0, 0.5, 3, and 6 h of reperfusion. We first determined the pathological outcomes after 7 days of recovery by histopathology, and then detected key components of programmed cell death pathway using immunocytochemistry coupled with confocal laser-scanning microscopy and Western blot analysis. The results show that the cytosolic cytochrome c increased mildly after reperfusion in nondiabetic samples. This increase was markedly enhanced in diabetic rats in both ischemic focus and penumbra. Subsequently, caspase-3 was activated and poly-ADP ribose polymerase (PARP) was cleaved. Our results suggest that activation of apoptotic cell death pathway may play a pivotal role in exaggerating brain damage in diabetic subjects. PMID- 12540625 TI - Effects of free fatty acids on glucose uptake and utilization in healthy women. AB - To study effects of sex on free fatty acid (FFA)-induced insulin resistance, we have examined the effects of acute elevations of plasma FFA levels on insulin stimulated total body glucose uptake in nine healthy young women. Euglycemic hyperinsulinemic (approximately 500 pmol/l) clamps were performed for 4 h with coinfusion of either lipid/heparin (L/H) to acutely raise plasma FFA levels (from approximately 600 to approximately 1,200 micro mol/l) or saline/glycerol to lower fatty acids (from approximately 600 to approximately 50 micro mol/l). L/H infusion inhibited insulin-stimulated glucose uptake (determined with [3 (3)H]glucose) and glycogen synthesis by 31 and 40%, respectively (P < 0.01), almost completely abolished insulin suppression of endogenous glucose production (EGP) (13.6 vs. 10.0 micro mol x kg(-1) x min(-1), NS), prevented the insulin induced increase in carbohydrate oxidation (8.1 vs. 7.4 micro mol x kg(-1) x min( 1), NS), and stimulated fat oxidation (from 3.6 to 5.1 micro mol x kg(-1) x min( 1), P < 0.01). These data showed that acute increases in plasma FFA levels inhibited the actions of insulin on glucose uptake, glycogen synthesis, and EGP in women to a degree similar to that previously reported in men. We conclude that at insulin and FFA levels in the postprandial range, women and men were susceptible to FFA-induced peripheral and hepatic insulin resistance. PMID- 12540626 TI - Demonstration of glycated insulin in human diabetic plasma and decreased biological activity assessed by euglycemic-hyperinsulinemic clamp technique in humans. AB - The presence and biological significance of circulating glycated insulin has been evaluated by high-pressure liquid chromatography (HPLC), electrospray ionization mass spectrometry (ESI-MS), radioimmunoassay (RIA), receptor binding, and hyperinsulinemic-euglycemic clamp techniques. ESI-MS analysis of an HPLC-purified plasma pool from four male type 2 diabetic subjects (HbA(1c) 8.1 +/- 0.2%, plasma glucose 8.7 +/- 1.3 mmol/l [means +/- SE]) revealed two major insulin-like peaks with retention times of 14-16 min. After spectral averaging, the peak with retention time of 14.32 min exhibited a prominent triply charged (M+3H)(3+) species at 1,991.1 m/z, representing monoglycated insulin with an intact M(r) of 5,970.3 Da. The second peak (retention time 15.70 min) corresponded to native insulin (M(r) 5,807.6 Da), with the difference between the two peptides (162.7 Da) representing a single glucitol adduct (theoretical 164 Da). Measurement of glycated insulin in plasma of type 2 diabetic subjects by specific RIA gave circulating levels of 10.1 +/- 2.3 pmol/l, corresponding to approximately 9% total insulin. Biological activity of pure synthetic monoglycated insulin (insulin B-chain Phe(1)-glucitol adduct) was evaluated in seven overnight-fasted healthy nonobese male volunteers using two-step euglycemic-hyperinsulinemic clamps (2 h at 16.6 micro g x kg(-1) x min(-1), followed by 2 h at 83.0 micro g x kg(-1) x min(-1); corresponding to 0.4 and 2.0 mU x kg(-1) x min(-1)). At the lower dose, the exogenous glucose infusion rates required to maintain euglycemia during steady state were significantly lower with glycated insulin (P < 0.01) and approximately 70% more glycated insulin was required to induce a similar rate of insulin-mediated glucose uptake. Maximal responses at the higher rates of infusion were similar for glycated and control insulin. Inhibitory effects on endogenous glucose production, insulin secretion, and lipolysis, as indicated by measurements of C-peptide, nonesterified free fatty acids, and glycerol, were also similar. Receptor binding to CHO-T cells transfected with human insulin receptor and in vivo metabolic clearance revealed no differences between glycated and native insulin, suggesting that impaired biological activity is due to a postreceptor effect. The present demonstration of glycated insulin in human plasma and related impairment of physiological insulin-mediated glucose uptake suggests a role for glycated insulin in glucose toxicity and impaired insulin action in type 2 diabetes. PMID- 12540627 TI - Comparative trial of N-acetyl-cysteine, taurine, and oxerutin on skin and kidney damage in long-term experimental diabetes. AB - This study analyzes the effect of chronic treatment with different antioxidants (N-acetyl-cysteine [NAC], taurine, a combination of NAC and taurine, and oxerutin) on long-term experimental diabetes induced by streptozotocin in rats. Glycoxidative damage was evaluated in the skin; glomerular structural changes were studied with morphometry and immunohistochemistry. Oxerutin treatment and the combined NAC plus taurine treatment resulted in reduced accumulation of collagen-linked fluorescence in skin in comparison with untreated diabetic rats. All treatments except taurine reduced glomerular accumulation of N(epsilon) (carboxymethyl)lysine and protected against the increase in glomerular volume typical of diabetes; furthermore, the apoptosis rate was significantly decreased and the glomerular cell density was better preserved. Glycoxidative markers in the skin turned out to be good indicators of the glomerular condition. The findings that emerged from our study support the hypothesis that glomerular damage in diabetes can be prevented or at least attenuated by supplementation with specific antioxidants. Treatment with oxerutin and combined treatment with NAC plus taurine gave the most encouraging results, whereas the results of taurine-only treatment were either negligible or negative and therefore suggest caution in the use of this molecule in single-drug treatment courses. PMID- 12540628 TI - A role for the polyol pathway in the early neuroretinal apoptosis and glial changes induced by diabetes in the rat. AB - We tested the hypothesis that the apoptosis of inner retina neurons and increased expression of glial fibrillary acidic protein (GFAP) observed in the rat after a short duration of diabetes are mediated by polyol pathway activity. Rats with 10 weeks of streptozotocin-induced diabetes and GHb levels of 16 +/- 2% (mean +/- SD) showed increased retinal levels of sorbitol and fructose, attenuation of GFAP immunostaining in astrocytes, appearance of prominent GFAP expression in Muller glial cells, and a fourfold increase in the number of apoptotic neurons when compared with nondiabetic rats. The cells undergoing apoptosis were immunoreactive for aldose reductase. Sorbinil, an inhibitor of aldose reductase, prevented all abnormalities. Intensive insulin treatment also prevented most abnormalities, despite reducing GHb only to 12 +/- 1%. Diabetic mice, known to have much lower aldose reductase activity in other tissues when compared with rats, did not accumulate sorbitol and fructose in the retina and were protected from neuronal apoptosis and GFAP changes in the presence of GHb levels of 14 +/- 2%. This work documents discrete cellular consequences of polyol pathway activity in the retina, and it suggests that activation of the pathway and "retinal neuropathy" require severe hyperglycemia and/or high activity of aldose reductase. These findings have implications for how to evaluate the role of the polyol pathway in diabetic retinopathy. PMID- 12540629 TI - Protein kinase C beta inhibition attenuates the progression of experimental diabetic nephropathy in the presence of continued hypertension. AB - In addition to hyperglycemia, hypertension and the renin-angiotensin system have been consistently implicated in the pathogenesis of diabetic nephropathy. Each of these pathogenetic factors may induce changes in cellular function by a common intracellular signaling pathway, the activation of protein kinase C (PKC) beta. The present study thus sought to determine the in vivo effect of PKC beta inhibition in experimental diabetic nephropathy in the setting of continued hyperglycemia, hypertension, and activation of the RAS. Studies were conducted in the (mRen-2)27 rat, a rodent that is transgenic for the entire mouse renin gene (Ren-2) and develops many of the structural, functional, and molecular characteristics of human diabetic nephropathy when experimental diabetes is induced with streptozotocin (STZ). Six-week-old female Ren-2 rats received an injection of STZ or vehicle and were maintained for 6 months. Within 24 h, diabetic rats were further randomized to receive treatment with the specific PKC beta inhibitor, LY333531, admixed in diet (10 mg x kg(-1) x d(-1)) or no treatment (n = 8/group). Diabetic rats developed albuminuria, glomerulosclerosis, and tubulointerstitial fibrosis with a concomitant increase in transforming growth factor-beta (TGF-beta). Western blot analysis demonstrated increased PKC beta in diabetic animals, localized by immunofluorescence to the glomerular mesangium. In vivo inhibition of PKC beta with LY333531 led to a reduction in albuminuria, structural injury, and TGF-beta expression, despite continued hypertension and hyperglycemia. PMID- 12540630 TI - Glucose-induced phosphatidylinositol 3-kinase and mitogen-activated protein kinase-dependent upregulation of the platelet-derived growth factor-beta receptor potentiates vascular smooth muscle cell chemotaxis. AB - The aim of this study was to investigate the effects of elevated D-glucose concentrations on vascular smooth muscle cell (VSMC) expression of the platelet derived growth factor (PDGF)beta receptor and VSMC migratory behavior. Immunoprecipitation, immunofluorescent staining, and RT-PCR of human VSMCs showed that elevated D-glucose induced an increase in the PDGFbeta receptor that was inhibited by phosphatidylinositol 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) pathway inhibitors. Exposure to 25 mmol/l D-glucose (HG) induced increased phosphorylation of protein kinase B (PKB) and extracellular-regulated kinase (ERK). All HG chemotaxis assays (with either 10 days' preincubation in HG or no preincubation) in a FCS or PDGF-BB gradient showed positive chemotaxis, whereas those in 5 mmol/l D-glucose did not. Assays were also run with concentrations ranging from 5 to 25 mmol/l D-glucose. Chemotaxis was induced at concentrations > or =9 mmol/l D-glucose. An anti-PDGFbeta receptor antibody inhibited glucose-potentiated VSMC chemotaxis, as did the inhibitors for the PI3K and MAPK pathways. This study has shown that small increases in D-glucose concentration, for a short period, increase VSMC expression of the PDGFbeta receptor and VSMC sensitivity to chemotactic factors in serum, leading to altered migratory behavior in vitro. It is probable that similar processes occur in vivo with glucose-enhanced chemotaxis of VSMCs, operating through PDGFbeta receptor operated pathways, contributing to the accelerated formation of atheroma in diabetes. PMID- 12540631 TI - Evidence for a novel TGF-beta1-independent mechanism of fibronectin production in mesangial cells overexpressing glucose transporters. AB - Recent experimental work indicates that the hyperglycemia-induced increase in mesangial matrix production, which is a hallmark in the development of diabetic nephropathy, is mediated by increased expression of GLUT1. Mesangial cells stably transfected with human GLUT1 mimic the effect of hyperglycemia on the production of the extracellular matrix proteins, particularly fibronectin, when cultured under normoglycemic conditions. Our investigation of the molecular mechanism of this effect has revealed that the enhanced fibronectin production was not mediated by the prosclerotic cytokine transforming growth factor (TGF)-beta1. We found markedly increased nuclear content in Jun proteins, leading to enhanced DNA binding activity of activating protein 1 (AP-1). AP-1 inhibition reduced fibronectin production in a dosage-dependent manner. Moreover, inhibition of classic protein kinase C (PKC) isoforms prevented both the activation of AP-1 and the enhanced fibronectin production. In contrast to mesangial cells exposed to high glucose, no activation of the hexosamine biosynthetic, p38, or extracellular signal-related kinase 1 and 2 mitogen-activated protein kinase pathways nor any increase in TGF-beta1 synthesis could be detected, which could be explained by the absence of oxidative stress in cells transfected with the human GLUT1 gene. Our data indicate that increased glucose uptake and metabolism induce PKC dependent AP-1 activation that is sufficient for enhanced fibronectin production, but not for increased TGF-beta1 expression. PMID- 12540632 TI - Amelioration of sensory nerve dysfunction by C-Peptide in patients with type 1 diabetes. AB - Studies have demonstrated that proinsulin C-peptide stimulates the activities of Na(+),K(+)-ATPase and endothelial nitric oxide synthase, both of which are enzyme systems of importance for nerve function and known to be deficient in type 1 diabetes. The aim of this randomized double-blind placebo-controlled study was to investigate whether C-peptide replacement improves nerve function in patients with type 1 diabetes. Forty-nine patients without symptoms of peripheral neuropathy were randomized to either 3 months of treatment with C-peptide (600 nmol/24 h, four doses s.c.) or placebo. Forty-six patients (15 women and 31 men, aged 29 years, diabetes duration 10 years, and HbA(1c) 7.0%) completed the study. Neurological and neurophysiological measurements were performed before and after 6 and 12 weeks of treatment. At baseline the patients showed reduced nerve conduction velocities in the sural nerve (sensory nerve conduction velocity [SCV]: 50.9 +/- 0.70 vs. 54.2 +/- 1.2 m/s, P < 0.05) and peroneal nerve (motor nerve conduction velocity: 45.7 +/- 0.55 vs. 53.5 +/- 1.1 m/s, P < 0.001) compared with age-, height-, and sex-matched control subjects. In the C-peptide treated group there was a significant improvement in SCV amounting to 2.7 +/- 0.85 m/s (P < 0.05 compared with placebo) after 3 months of treatment, representing 80% correction of the initial reduction in SCV. The change in SCV was accompanied by an improvement in vibration perception in the patients receiving C-peptide (P < 0.05 compared with placebo), whereas no significant change was detectable in cold or heat perception. In conclusion, C-peptide administered for 3 months as replacement therapy to patients with early signs of diabetic neuropathy ameliorates nerve dysfunction. PMID- 12540633 TI - Microvascular alterations in diabetic mice correlate with level of hyperglycemia. AB - Vascular alterations are the most common causes of morbidity and mortality in diabetic patients. Despite the impact of endothelial dysfunction on microcirculatory properties, little is known about the endothelial cell alteration during the development of diabetes and its correlation to the metabolic situation. For that reason we continuously monitored in vivo functional and morphological alterations of the microvasculature in hyperglycemic and hyperinsulinemic transgenic UCP1/DTA mice with brown fat deficiency, using a dorsal skin-fold chamber preparation and fluorescence microscopy. UCP1/DTA mice showed a dramatic decrease in vascular density due to a remarkable reduction of small vessels. Vascular permeability and leukocyte endothelial interactions (LEIs) significantly increased. The extent of vascular alteration correlated with the extent of metabolic dysfunction. Decreased tissue perfusion observed in UCP1/DTA mice might play a role in impaired wound healing observed in diabetes. The increased permeability in subcutaneous tissue may serve as predictor of vascular changes in early stages of diabetes. The increased LEI and serum tumor necrosis factor-alpha levels, which mirror the inflammatory process, support the growing evidence of the inflammatory component of diabetic disease. The results suggest that anti-inflammatory strategies might be able to prevent vascular deterioration in early stages of diabetes. Further investigations are required to evaluate the benefit of such therapeutic strategies. PMID- 12540634 TI - Genome-wide and fine-mapping linkage studies of type 2 diabetes and glucose traits in the Old Order Amish: evidence for a new diabetes locus on chromosome 14q11 and confirmation of a locus on chromosome 1q21-q24. AB - We conducted a genome scan using a 10-cM map to search for genes linked to type 2 diabetes in 691 individuals from a founder population, the Old Order Amish. We then saturated two regions on chromosomes 1 and 14 showing promising linkage signals with additional markers to produce a approximately 2-cM map for fine mapping. Analyses of both discrete traits (type 2 diabetes and the composite trait of type 2 diabetes and/or impaired glucose homeostasis [IGH]), and quantitative traits (glucose levels during a 75-g oral glucose challenge, designated glucose 0-180 and HbA(1c)) were performed. We obtained significant evidence for linkage to type 2 diabetes in a novel region on chromosome 14q11 (logarithm of odds [LOD] for diabetes = 3.48, P = 0.00005). Furthermore, we observed evidence for the existence of a diabetes-related locus on chromosome 1q21-q24 (LOD for type 2 diabetes/IGH = 2.35, P = 0.0008), a region shown to be linked to diabetes in several other studies. Suggestive evidence for linkage to glucose traits was observed on three other regions: 14q11-q13 (telomeric to that above with LOD = 1.82-1.85 for glucose 150 and 180), 1p31 (LOD = 1.28-2.30 for type 2 diabetes and glucose 120-180), and 18p (LOD = 3.07, P = 0.000085 for HbA(1c) and LOD = 1.50 for glucose 0). In conclusion, our findings provide evidence that type 2 diabetes susceptibility genes reside on chromosomes 1, 14, and 18. PMID- 12540635 TI - The C-174G promoter polymorphism of the IL-6 gene affects energy expenditure and insulin sensitivity. AB - Interleukin-6 (IL-6) is a pleiotropic cytokine expressed in many tissues. IL-6 null mice show low energy expenditure, but the effect of the variants of the IL-6 gene on energy expenditure has not been previously studied in humans. Therefore, we investigated the effect of the C-174G promoter polymorphism of the IL-6 gene on energy expenditure, measured by indirect calorimetry in healthy Finnish subjects (n = 124). We also measured insulin sensitivity by the hyperinsulinemic euglycemic clamp. Subjects with the C-174C genotype of the IL-6 gene had significantly lower energy expenditure than subjects with the G-174C or G-174G genotypes both in fasting (CC 13.68 +/- 1.98, CG 14.73 +/- 1.57, GG 14.81 +/- 2.01 kcal x kg(-1) x min(-1); P = 0.012) and during the euglycemic hyperinsulinemic clamp (CC 15.24 +/- 2.05, CG 16.62 +/- 2.06, GG 16.66 +/- 2.50 kcal x kg(-1) x min(-1); P = 0.007). Moreover, subjects homozygous for the C allele had lower rates of whole-body glucose uptake than carriers of the G allele (CC 50.95 +/- 13.91, CG 59.40 +/- 14.17, GG 59.21 +/- 15.93 micro mol x kg(-1) x min(-1); P = 0.016). The rates of both oxidative (P = 0.013) and nonoxidative (P = 0.016) glucose disposal were significantly affected by the IL-6 promoter polymorphism. In conclusion, the C-174C promoter polymorphism of the IL-6 gene influences energy expenditure and insulin sensitivity in healthy normoglycemic subjects. Whether this polymorphism is a risk factor for obesity or type 2 diabetes can be estimated only in prospective population-based studies. PMID- 12540636 TI - Systematic search for single nucleotide polymorphisms in the FOXC2 gene: the absence of evidence for the association of three frequent single nucleotide polymorphisms and four common haplotypes with Japanese type 2 diabetes. AB - FOXC2, a forkhead/winged helix transcription factor, represents a promising candidate gene for type 2 diabetes since transgenic mice that specifically overexpress this gene in adipocytes are lean and insulin sensitive. To determine whether there are single nucleotide polymorphisms (SNPs) in this gene that are associated with type 2 diabetes, sequences of the coding and approximately 1 kb of 5' flanking regions in 24 Japanese type 2 diabetic subjects were initially analyzed using PCR direct sequencing, and the regions containing the identified polymorphisms were then examined. In 200 control subjects, three frequent SNPs were found (g. -512C>T [32.3%] and -350G>T [13.0%] in the 5' flanking region and +1548C>T [10.0%] in the 3' flanking region). Linkage disequilibria were found between all three pairs of these SNPs. Of the eight possible haplotypes defined by these SNPs, only four were found. When the frequencies of these SNPs and the four common haplotypes between 195 type 2 diabetic and 200 control subjects were compared, no association was evident. The +898C>T (Pro300Ser), +907C>A (Leu303Met), 1167_1169delCCA (389delHis), and +1251C>A (Ala417Ala) identified in the coding region were rare, although +907C>A could be higher in type 2 diabetic subjects (1.5%) than in control subjects (0.3%). Thus, the SNPs identified in the FOXC2 gene are unlikely to have major effects on susceptibility to Japanese type 2 diabetes. PMID- 12540637 TI - Large-scale association studies of variants in genes encoding the pancreatic beta cell KATP channel subunits Kir6.2 (KCNJ11) and SUR1 (ABCC8) confirm that the KCNJ11 E23K variant is associated with type 2 diabetes. AB - The genes ABCC8 and KCNJ11, which encode the subunits sulfonylurea receptor 1 (SUR1) and inwardly rectifying potassium channel (Kir6.2) of the beta-cell ATP sensitive potassium (K(ATP)) channel, control insulin secretion. Common polymorphisms in these genes (ABCC8 exon 16-3t/c, exon 18 T/C, KCNJ11 E23K) have been variably associated with type 2 diabetes, but no large ( approximately 2,000 subjects) case-control studies have been performed. We evaluated the role of these three variants by studying 2,486 U.K. subjects: 854 with type 2 diabetes, 1,182 population control subjects, and 150 parent-offspring type 2 diabetic trios. The E23K allele was associated with diabetes in the case-control study (odds ratio [OR] 1.18 [95% CI 1.04-1.34], P = 0.01) but did not show familial association with diabetes. Neither the exon 16 nor the exon 18 ABCC8 variants were associated with diabetes (1.04 [0.91-1.18], P = 0.57; 0.93 [0.71-1.23], P = 0.63, respectively). Meta-analysis of all case-control data showed that the E23K allele was associated with type 2 diabetes (K allele OR 1.23 [1.12-1.36], P = 0.000015; KK genotype 1.65 [1.34-2.02], P = 0.000002); but the ABCC8 variants were not associated. Our results confirm that E23K increases risk of type 2 diabetes and show that large-scale association studies are important for the identification of diabetes susceptibility alleles. PMID- 12540638 TI - The E23K variant of Kir6.2 associates with impaired post-OGTT serum insulin response and increased risk of type 2 diabetes. AB - The E23K polymorphism of the pancreatic beta-cell ATP-sensitive K(+) (K(ATP)) channel subunit Kir6.2 (KCNJ11) is associated with type 2 diabetes in whites, and a recent in vitro study of the E23K variant suggests that the association to diabetes might be explained by a slight inhibition of serum insulin release. In a study comprising 519 unrelated glucose-tolerant subjects, we addressed the question as to whether the E23K variant was related to reduced serum insulin release during an oral glucose tolerance test (OGTT). Furthermore, the polymorphism was examined in a case-control study comprising 803 type 2 diabetic patients and 862 glucose-tolerant control subjects. The E23K variant was associated with significant reductions in the insulinogenic index (P = 0.022) and serum insulin levels under the response curve during an OGTT (0-120 min) (P = 0.014) as well as with an increase in BMI (P = 0.013). In the present study, the association of the E23K polymorphism with type 2 diabetes was not significant (P = 0.26). However, the K23K genotype significantly associated with type 2 diabetes in a meta-analysis of white case and control subjects (n = 2,824, odds ratio [OR] 1.49, P = 0.00022). In conclusion, the widespread E23K polymorphism may have a diabetogenic effect by impairing glucose-induced insulin release and increasing BMI. PMID- 12540639 TI - Magnesium sulfate for preeclampsia. PMID- 12540640 TI - Environmental lead exposure and progression of chronic renal diseases in patients without diabetes. AB - BACKGROUND: Previous research suggests that environmental lead exposure correlates with age-related decreases in renal function. METHODS: Two hundred two patients with chronic renal insufficiency (indicated by a serum creatinine level between 1.5 mg per deciliter and 3.9 mg per deciliter) who had a normal total body lead burden and no history of exposure to lead were observed for 24 months. After the observation period, 64 subjects with an elevated body lead burden were randomly assigned to the chelation control groups. For three months, the patients in the chelation group received lead-chelation therapy with calcium disodium EDTA, and the control group received placebo. During the ensuing 24 months, repeated chelation therapy was administered weekly to 32 patients with high normal body lead burdens (at least 80 microg but less than 600 microg) unless on repeated testing the body lead burden fell below 60 microg; the other 32 patients served as controls and received weekly placebo infusions for 5 weeks every 6 months. The primary end point was an increase in the serum creatinine level to 1.5 times the base-line value during the observation period. A secondary end point was the change in renal function during the intervention period. RESULTS: The primary end point occurred in 24 patients during the observation period; the serum creatinine levels and body lead burden at base line were the most important risk factors. The glomerular filtration rate improved significantly by the end of the 27-month intervention period in patients receiving chelation therapy: the mean (+/-SD) change in the glomerular filtration rate in the patients in the chelation group was 2.1+/-5.7 ml per minute per 1.73 m2 of body-surface area, as compared with -6.0+/-5.8 ml per minute per 1.73 m2 of body-surface area in the controls (P<0.001). The rate of decline in the glomerular filtration rate in the chelation group was also lower than that in the controls during the 24-month period of repeated chelation therapy or placebo. CONCLUSIONS: Low-level environmental lead exposure may accelerate progressive renal insufficiency in patients without diabetes who have chronic renal disease. Repeated chelation therapy may improve renal function and slow the progression of renal insufficiency. PMID- 12540641 TI - Serum retinol levels and the risk of fracture. AB - BACKGROUND: Although studies in animals and epidemiologic studies have indicated that a high vitamin A intake is associated with increased bone fragility, no biologic marker of vitamin A status has thus far been used to assess the risk of fractures in humans. METHODS: We enrolled 2322 men, 49 to 51 years of age, in a population-based, longitudinal cohort study. Serum retinol and beta carotene were analyzed in samples obtained at enrollment. Fractures were documented in 266 men during 30 years of follow-up. Cox regression analysis was used to determine the risk of fracture according to the serum retinol level. RESULTS: The risk of fracture was highest among men with the highest levels of serum retinol. Multivariate analysis of the risk of fracture in the highest quintile for serum retinol (>75.62 microg per deciliter [2.64 micromol per liter]) as compared with the middle quintile (62.16 to 67.60 microg per deciliter [2.17 to 2.36 micromol per liter]) showed that the rate ratio was 1.64 (95 percent confidence interval, 1.12 to 2.41) for any fracture and 2.47 (95 percent confidence interval, 1.15 to 5.28) for hip fracture. The risk of fracture was further increased within the highest quintile for serum retinol. Men with retinol levels in the 99th percentile (>103.12 microg per deciliter [3.60 micromol per liter]) had an overall risk of fracture that exceeded the risk among men with lower levels by a factor of seven (P<0.001). The level of serum beta carotene was not associated with the risk of fracture. CONCLUSIONS: Our findings, which are consistent with the results of studies in animals, as well as in vitro and epidemiologic dietary studies, suggest that current levels of vitamin A supplementation and food fortification in many Western countries may need to be reassessed. PMID- 12540642 TI - Effect of left ventricular outflow tract obstruction on clinical outcome in hypertrophic cardiomyopathy. AB - BACKGROUND: The influence of left ventricular outflow tract obstruction on the clinical outcome of hypertrophic cardiomyopathy remains unresolved. METHODS: We assessed the effect of outflow tract obstruction on morbidity and mortality in a large cohort of patients with hypertrophic cardiomyopathy who were followed for a mean (+/-SD) of 6.3+/-6.2 years. RESULTS: Of the 1101 consecutive patients, 273 (25 percent) had obstruction of left ventricular outflow under basal (resting) conditions with a peak instantaneous gradient of at least 30 mm Hg. A total of 127 patients (12 percent) died of hypertrophic cardiomyopathy, and 216 surviving patients (20 percent) had severe, disabling symptoms of progressive heart failure (New York Heart Association [NYHA] functional class III or IV). The overall probability of death related to hypertrophic cardiomyopathy was significantly greater among patients with outflow tract obstruction than among those without obstruction (relative risk, 2.0; P=0.001). The risk of progression to NYHA class III or IV or death specifically from heart failure or stroke was also greater among patients with obstruction (relative risk, 4.4; P<0.001), particularly among patients 40 years of age or older (P<0.001). Age-adjusted multivariate analysis confirmed that outflow tract obstruction was independently associated with an increased risk of both death related to hypertrophic cardiomyopathy (relative risk, 1.6; P=0.02) and progression to NYHA class III or IV or death from heart failure or stroke (relative risk, 2.7; P<0.001). The likelihood of severe symptoms and death related to outflow tract obstruction did not increase as the gradient increased above the threshold of 30 mm Hg. CONCLUSIONS: In patients with hypertrophic cardiomyopathy, left ventricular outflow tract obstruction at rest is a strong, independent predictor of progression to severe symptoms of heart failure and of death. PMID- 12540643 TI - A comparison of magnesium sulfate and nimodipine for the prevention of eclampsia. AB - OBJECTIVE: Magnesium sulfate may prevent eclampsia by reducing cerebral vasoconstriction and ischemia. Nimodipine is a calcium-channel blocker with specific cerebral vasodilator activity. Our objective was to determine whether nimodipine is more effective than magnesium sulfate for seizure prophylaxis in women with severe preeclampsia. METHODS: We conducted an unblinded, multicenter trial in which 1650 women with severe preeclampsia were randomly assigned to receive either nimodipine (60 mg orally every 4 hours) or intravenous magnesium sulfate (given according to the institutional protocol) from enrollment until 24 hours post partum. High blood pressure was controlled with intravenous hydralazine as needed. The primary outcome measure was the development of eclampsia, as defined by a witnessed tonic-clonic seizure. RESULTS: Demographic and clinical characteristics were similar in the two groups. The women who received nimodipine were more likely to have a seizure than those who received magnesium sulfate (21 of 819 [2.6 percent] vs. 7 of 831 [0.8 percent], P=0.01). The adjusted risk ratio for eclampsia associated with nimodipine, as compared with magnesium sulfate, was 3.2 (95 percent confidence interval, 1.1 to 9.1). The antepartum seizure rates did not differ significantly between groups, but the nimodipine group had a higher rate of postpartum seizures (9 of 819 [1.1 percent] vs. 0 of 831, P=0.01). There were no significant differences in neonatal outcome between the two groups. More women in the magnesium sulfate group than in the nimodipine group needed hydralazine to control blood pressure (54.3 percent vs. 45.7 percent, P<0.001). CONCLUSIONS: Magnesium sulfate is more effective than nimodipine for prophylaxis against seizures in women with severe preeclampsia. PMID- 12540644 TI - Ectopic luteinizing hormone secretion and anovulation. PMID- 12540645 TI - Images in clinical medicine. Broncholithiasis. PMID- 12540646 TI - Regional anesthesia and analgesia for labor and delivery. PMID- 12540647 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 3-2003. A 36-year-old man with renal failure, hypertension, and neurologic abnormalities. PMID- 12540649 TI - Increased body lead burden--cause or consequence of chronic renal insufficiency? PMID- 12540650 TI - Hypervitaminosis A and fractures. PMID- 12540651 TI - A family of WASPs. PMID- 12540652 TI - Is the match illegal? PMID- 12540653 TI - Thrombolytic therapy in patients with submassive pulmonary embolism. PMID- 12540654 TI - Inhibition of tumor necrosis factor alpha and ankylosing spondylitis. PMID- 12540655 TI - Antimicrobial peptides in the skin. PMID- 12540656 TI - Otitis media. PMID- 12540657 TI - Helicobacter pylori. PMID- 12540658 TI - Changing health insurance trends. PMID- 12540659 TI - Increased prevalence of sleep-disordered breathing among professional football players. PMID- 12540660 TI - Driving fatalities on Super Bowl Sunday. PMID- 12540662 TI - Cellular mechanisms of osteolysis. PMID- 12540664 TI - Why we should collect outcomes data. PMID- 12540663 TI - Polyethylene damage in total knees and use of highly crosslinked polyethylene. PMID- 12540665 TI - Current status of revision total knee arthroplasty: how do we assess results? PMID- 12540666 TI - Modes of failure and preoperative evaluation. PMID- 12540667 TI - Intraoperative assessment in revision total knee arthroplasty. PMID- 12540668 TI - Soft-tissue balance in revision total knee arthroplasty. PMID- 12540669 TI - Implant selection in revision total knee arthroplasty. PMID- 12540670 TI - Strategies of stem fixation and the role of supplemental bone graft in revision total knee arthroplasty. PMID- 12540671 TI - Salvage procedures for failed total knee arthroplasty. PMID- 12540672 TI - The patella in revision total knee arthroplasty. PMID- 12540673 TI - Complications after revision total knee arthroplasty. PMID- 12540674 TI - Diagnosis and management of infection after total knee arthroplasty. PMID- 12540678 TI - The Argentinean paradox: the case of contradictory child malnutrition epidemics. PMID- 12540679 TI - Building health policy for healthy living in the city. PMID- 12540680 TI - Theory in carcinogenesis and epidemiology. PMID- 12540681 TI - What characterises a useful concept of causation in epidemiology? AB - It has recently been suggested that epidemiologists should avoid thinking of causes in deterministic terms. This would mean giving up the component-cause model in its original form. A model that has provided important contributions as to how we develop hypotheses, design our study, analyse data, interpret and communicate our results. The component-cause model has considerably more to offer than a simple probabilistic concept. What a causal model has to offer to the advancement of the discipline is equally important as the concept itself. It has been said that we should not hunt "the Holy Grail" (that is, determinism), if it does not exist. This line of reasoning neglects the fact that the "hunting" is more important than the "finding". PMID- 12540682 TI - An evolutionary paradigm for carcinogenesis? AB - Mutations seem to be only one of the mechanisms involved in carcinogenesis; selection of mutated clones is a second crucial mechanism. An evolutionary (darwinian) theory of carcinogenesis can be useful to explain some contradictory observations of epidemiology, and to provide a common theoretical framework for carcinogenesis. In both the selection of species and in carcinogenesis (selection of mutated cells), mutation and selection can be interpreted as necessary and insufficient causes. Selection presupposes competition among clones-that is, survival advantage of the mutated species; without selective forces a mutation is mute, while the lack of mutations makes selective advantage impossible. The identification of carcinogen related fingerprints is ambiguous: it can suggest both a genuine mutational hotspot left by the carcinogenic stimulus (like in tobacco related p53 mutations), and selective advantage of clones whose mutations seem to be not exposure specific (like in the case of aflatoxin). We present several examples of exposures that can increase the risk of cancer in humans not via mutations but through a putative mechanism of clone selection. PMID- 12540683 TI - The challenges of evaluating environmental interventions to increase population levels of physical activity: the case of the UK National Cycle Network. PMID- 12540684 TI - Watching the Games: public health surveillance for the Sydney 2000 Olympic Games. AB - STUDY OBJECTIVE: To describe the development of the public health surveillance system for the Sydney 2000 Olympic Games; document its major findings; and discuss the implications for public health surveillance for future events. DESIGN: Planning for the system took almost three years. Its major components included increased surveillance of communicable diseases; presentations to sentinel emergency departments; medical encounters at Olympic venues; cruise ship surveillance; environmental and food safety inspections; surveillance for bioterrorism; and global epidemic intelligence. A daily report integrated data from all sources. SETTING: Sydney, Australia. Surveillance spanned the period 28 August to 4 October 2000. PARTICIPANTS: Residents of Sydney, athletes and officials, Australian and international visitors. MAIN RESULTS: No outbreaks of communicable diseases were detected. There were around 5% more presentations to Sydney emergency departments than in comparable periods in other years. Several incidents detected through surveillance, including injuries caused by broken glass, and a cluster of presentations related to the use of the drug ecstasy, prompted further action. CONCLUSIONS: Key elements in the success of public health surveillance for the Games included its careful planning, its comprehensive coverage of public health issues, and its timely reporting and communication processes. Future systems need to be flexible enough to detect the unexpected. PMID- 12540685 TI - Previous consultation and self reported health status as predictors of future demand for primary care. AB - STUDY OBJECTIVE: There has been little prospective investigation of what predicts general practice consultation. The objective of this study was to investigate the extent to which previous primary care consultation and self reported health status are predictors of future primary care consultation. DESIGN: Population based cohort study in two phases. Firstly, a baseline survey (1995/96) to identify the cohort and to obtain self reported health status using the UK census limiting long term illness (LLI) question and the Short Form-36 (SF-36) health profile. Secondly, analysis of general practice medical records for two years (1994/1995) before the survey and for two years (1997/1998) after the survey. Analysis was performed on: (a) all contacts coded by the GP, (b) the subgroup of contacts given a diagnostic morbidity code by the GP. SETTING: One general practice in North Staffordshire, UK. PARTICIPANTS: 738 survey respondents who had consented to viewing of medical records including all those who reported LLI together with an age-gender matched control group of those who reported no LLI. MAIN RESULTS: High frequency consulters in 1994/95 were more likely than non consulters or average consulters in that year to be high consulters in 1997/98 (odds ratio 5.6, 95% confidence interval 3.82 to 8.25, for all contacts; 4.4 for diagnostic coded consultations). Self reported role disability and physical limitation from the SF-36 at baseline increased the probability of being a future high consulter but the effects were weaker than for previous consultation. Previous consultation within a diagnostic group was the main predictor for future consultation within that group with weaker but significant prediction by self reported health status. CONCLUSIONS: Reliable morbidity coding in general practice provides the best available basis for predicting future demand in primary care. Self reported health status survey instruments add to this information but on their own are weaker predictors of future consultation. PMID- 12540686 TI - Social capital in a changing society: cross sectional associations with middle aged female and male mortality rates. AB - OBJECTIVES: Social capital has been linked to self rated health and mortality rates. The authors examined the relations between measures of social capital and male/female mortality rates across counties in Hungary. DESIGN: Cross sectional, ecological study. SETTING: 20 counties of Hungary. PARTICIPANTS AND METHODS: 12,640 people were interviewed in 1995 (the "Hungarostudy II" survey), representing the Hungarian population according to sex, age, and county. Social capital was measured by three indicators: lack of social trust, reciprocity between citizens, and help received from civil organisations. Covariates included county GDP, personal income, education, unemployment, smoking, and alcohol spirit consumption. MAIN OUTCOME MEASURE: Gender specific mortality rates were calculated for the middle aged population (45-64 years) in the 20 counties of Hungary. RESULTS: All of the social capital variables were significantly associated with middle age mortality, but levels of mistrust showed the strongest association. Several gender differences were observed, namely male mortality rates were more closely associated with lack of help from civic organisations, while female mortality rates were more closely connected with perceptions of reciprocity. CONCLUSION: There are gender differences in the relations of specific social capital indicators to mortality rates. At the same time, perceptions of social capital within each sex were associated with mortality rates in the opposite sex. PMID- 12540687 TI - Lethality of firearms relative to other suicide methods: a population based study. AB - OBJECTIVES: (1) To quantify lethality of firearms relative to other suicide methods, (2) to quantify the extent to which suicide mortality may be reduced by limiting access to firearms. METHODS: Data on suicides and hospitalised para suicides that occurred in the state of Illinois from 1990 to 1997 were combined. Total number of episodes for each suicide method was estimated as the sum of the number of suicides and the number of para-suicides for that method. Gender and suicide method were used as proxies for intention to die, and estimated lethality of suicide methods within method-gender groups (for example, male firearm users). Logistic regression was used to quantify the lethality of firearms relative to other suicide methods. Excess mortality associated with the use of firearms was estimated by conservatively assuming that in the absence of firearms the next most lethal suicide method would be used. RESULTS: From January 1990 to December 1997, among individuals 10 years or older in the state of Illinois, there were 37,352 hospital admissions for para-suicide and 10,287 completed suicides. Firearms are the most lethal suicide method. Episodes involving firearms are 2.6 times (95% CI 2.1 to 3.1) more lethal than those involving suffocation-the second most lethal suicide method. Preventing access to firearms can reduce the proportion of fatal firearms related suicides by 32% among minors, and 6.5% among adults. CONCLUSIONS: Limiting access to firearms is a potentially effective means of reducing suicide mortality. PMID- 12540688 TI - Income related inequalities in self assessed health in Britain: 1979-1995. AB - STUDY OBJECTIVE: To measure and decompose income related inequalities in self assessed health in England, Scotland, and Wales, 1979-1995. DESIGN: The relation between individual health and a non-linear transformation of equivalised income, allowing for sex, age, country, and year effects, was estimated by multiple regression. The share of health attributable to transformed income and the Gini coefficient for transformed income were calculated. Inequality in health was measured by the partial concentration index, which is the product of the Gini coefficient and the share of health attributable to transformed income. PARTICIPANTS AND SETTING: Representative annual samples of the adult population living in private households in Great Britain 1979-1995. The total analysed sample was 299,968 people. MAIN RESULTS: Pro-rich health inequality was largest in Wales and smallest in England over the period because the effect of increased income on health was greatest in Wales and least in England. In all three countries, pro-rich health inequality increased throughout the period. In the early 1980s this was primarily attributable to increases in income inequality. Thereafter the increased share of health attributable to income was the principal cause. CONCLUSIONS: Reductions in pro-rich health inequality can be achieved by reducing income inequality, reducing the effect of income on health, or both. PMID- 12540689 TI - Relation between body mass index and mortality in an unusually slim cohort. AB - STUDY OBJECTIVE: To investigate the relation between body mass index and mortality in an unusually slim cohort. DESIGN: Prospective cohort study. SETTING: United Kingdom. PARTICIPANTS: About 11,000 non-meat eaters and their meat eating friends and relatives, with a median age of 33 years were recruited between 1980 and 1984. More than 20% of the cohort had a self reported BMI below 20 kg/m(2) at recruitment. There were 195,000 person years of observation after a mean of 18.0 years of follow up. MAIN RESULTS: The characteristics of participants with a BMI below 18 kg/m(2) were favourable to a lower risk of cardiovascular disease. Nevertheless, these participants had an increased all cause death rate ratio (2.07, 95% CI 1.58 to 2.70) in comparison with participants who had a BMI between 20 and 22 kg/m(2). The death rate ratio for the slimmest category was also significantly increased for circulatory diseases (including ischaemic heart disease and cerebrovascular disease), respiratory diseases, and all other causes combined excluding all malignant neoplasms. This finding was consistent across a range of subgroups. CONCLUSIONS: Lean men and women (BMI <18 kg/m(2)) experience increased all cause mortality compared with those with a BMI between 20 and 22 kg/m(2). This pattern is not seen for cancer mortality, but is found for cardiovascular and respiratory diseases. It is important that public health messages regarding healthy eating are aimed at maintaining a healthy body weight rather than just "losing weight". PMID- 12540690 TI - Geographical variation in cardiovascular disease, risk factors, and their control in older women: British Women's Heart and Health Study. AB - OBJECTIVES: To measure the geographical variation in prevalence of cardiovascular disease, risk factors, and their control in a nationally representative sample of older British women. METHODS: Baseline survey using general practitioner record review, a self completed questionnaire, research nurse interview, and physical examination in a randomly selected sample of women aged 60-79 drawn from 23 towns in England, Scotland, and Wales. RESULTS: Of 7,173 women invited and eligible to participate, information was obtained on 4,286 (60%). One in five women had a doctor diagnosis of any one of myocardial infarction, angina, heart failure, stroke, or peripheral vascular disease. Fifty per cent of women were hypertensive, 12% smoked, and over one quarter were obese. Fifty per cent had a total cholesterol level greater than 6.5 mmol/l, though only 3% had low high density lipoprotein concentrations. Cardiovascular disease prevalence varied by geographical region being highest in Scotland: age adjusted prevalence (95% confidence intervals) 25.0% (21.5% to 28.8%) and lowest in South England: age adjusted prevalence (95% confidence intervals) 15.4% (13.5% to 17.6%). The geographical variations in cardiovascular disease prevalence were attenuated by adjustment for risk factors and socioeconomic position; further adjustment for health service use (as indicated by aspirin or statin use) reduced the differences further. However, variation remained even after full adjustment for these factors: odds ratio (95% confidence intervals) comparing Midlands and Wales to South England 1.15 (0.82 to 1.61) and comparing Scotland to South England 1.53 (1.08 to 2.14). Of women with cardiovascular disease, 12% were current smokers, a third had uncontrolled hypertension, a third were obese, and 90% had a blood cholesterol over 5 mmol/l. Only 41% were taking antiplatelet drugs and 22% were taking a statin. CONCLUSIONS: Older British women have a higher prevalence of cardiovascular disease and risk factors than previously documented. The workload consequences of attempting to control risk factors and ensure optimal secondary prevention for older British women are considerable. Geographical variations in cardiovascular disease prevalence in older women are somewhat, but not fully, explained by variations in major risk factors, socioeconomic position, and health service utilisation. PMID- 12540691 TI - Associations of height, leg length, and lung function with cardiovascular risk factors in the Midspan Family Study. AB - BACKGROUND: Taller people and those with better lung function are at reduced risk of coronary heart disease (CHD). Biological mechanisms for these associations are not well understood, but both measures may be markers for early life exposures. Some studies have shown that leg length, an indicator of pre-pubertal nutritional status, is the component of height most strongly associated with CHD risk. Other studies show that height-CHD associations are greatly attenuated when lung function is controlled for. This study examines (1) the association of height and the components of height (leg length and trunk length) with CHD risk factors and (2) the relative strength of the association of height and forced expiratory volume in one second (FEV(1)) with risk factors for CHD. SUBJECTS AND METHODS: Cross sectional analysis of data collected at detailed cardiovascular screening examinations of 1040 men and 1298 women aged 30-59 whose parents were screened in 1972-76. Subjects come from 1477 families and are members of the Midspan Family Study. SETTING: The towns of Renfrew and Paisley in the West of Scotland. RESULTS: Taller subjects and those with better lung function had more favourable cardiovascular risk factor profiles, associations were strongest in relation to FEV(1). Higher FEV(1) was associated with lower blood pressure, cholesterol, glucose, fibrinogen, white blood cell count, and body mass index. Similar, but generally weaker, associations were seen with height. These associations were not attenuated in models controlling for parental height. Longer leg length, but not trunk length, was associated with lower systolic and diastolic blood pressure. Longer leg length was also associated with more favourable levels of cholesterol and body mass index than trunk length. CONCLUSIONS: These findings provide indirect evidence that measures of lung development and pre-pubertal growth act as biomarkers for childhood exposures that may modify an individual's risk of developing CHD. Genetic influences do not seem to underlie height-CHD associations. PMID- 12540692 TI - Job strain, job demands, decision latitude, and risk of coronary heart disease within the Whitehall II study. AB - STUDY OBJECTIVES: To investigate the association between job strain and components of the job strain model and coronary heart disease (CHD) risk. DESIGN: Prospective cohort study (Whitehall II study). At the first phase of the study (1985-1988), data on self reported psychosocial work characteristics were collected from all participants. Participants were followed up until the end of phase 5 (1997-2000), with mean length of follow up of 11 years. SETTING: London based office staff in 20 civil service departments. PARTICIPANTS: 6,895 male and 3,413 female civil servants aged 35-55. OUTCOME MEASURES: Incident validated CHD. MAIN RESULTS: People with concurrent low decision latitude and high demands (job strain) were at the highest risk for CHD. High job demands, and, less consistently, low decision latitude, predicted CHD incidence. The effect of job strain on CHD incidence was strongest among younger workers, but there was no effect modification by social support at work, or employment grade. CONCLUSIONS: Job strain, high job demands, and, to some extent, low decision latitude, are associated with an increased risk of CHD among British civil servants. PMID- 12540694 TI - Are smoke-free policies good for business? PMID- 12540693 TI - Effect of work related variables on growth among working boys in Jordan. AB - OBJECTIVE: To study the effect of work related variables on the height, weight, and body mass index (BMI) of working boys aged 10-16 years. DESIGN: Cross sectional survey. SETTING: Irbid, Jarash, and North Jordan Valley in Jordan. PARTICIPANTS: 135 working boys aged 10-16 years and their mothers. OUTCOME MEASURES: Height for age z score; weight for age z score; BMI for age z score. RESULTS: In bivariate analysis, child's duration of work (r=-0.20), maternal height (r=0.26), and household per capita income (r=0.23) were significantly correlated with height for age z score and duration of work (r=-0.24), maternal height (r=0.24), and household per capita income (r=0.19) with weight for age z score. Duration of work (p<0.001), child's monthly income (p=0.044), maternal height (p=0.002), and household per capita income (p=0.005) were retained in the regression model fitted on height z score that explained 20.1% of the variance. The regression model fitted on weight z score explained 20.1% of the variance and duration of work (p<0.001), child's monthly income (p=0.022), household per capita income (p=0.017), and maternal height (p=0.004) were retained. Only duration of work (p=0.001) was retained in the model fitted on BMI for age z score. CONCLUSION: The results of this study suggest that the length of time children have been working and low monthly income have a detrimental effect on growth of working boys independent of the effects of low household per capita income and small maternal stature. Relevance of these findings for social policy and health care of working boys in Jordan and elsewhere is discussed. PMID- 12540695 TI - Intervention effects on youth tobacco use in the community intervention trial (COMMIT). PMID- 12540696 TI - Psychosocial determinants of health in social epidemiology. PMID- 12540697 TI - Social epidemiology: towards a better understanding of the field. PMID- 12540698 TI - Asthma and the westernization 'package'. PMID- 12540699 TI - Coronary disease and modern stress. PMID- 12540704 TI - The search for new risk factors for coronary heart disease: occupational therapy for epidemiologists? AB - The identification of the proximal causes of coronary heart disease (CHD) during the second half of the 20th century contributed to the prevention of premature CHD and the extension of life expectancy in middle-aged and older people in many wealthy countries. These major CHD risk factors-high blood cholesterol, high blood pressure, cigarette smoking and physical inactivity-satisfy public health criteria of causality. Strong epidemiological evidence suggests that they explain at least 75% of new cases of CHD. However, the search for 'new' or 'emerging' CHD risk factors continues, partly justified by a myth that minimizes the contribution of the major risk factors. The public health criteria of causality were applied to the following proposed new risk factors: thrombotic factors and serum homocysteine levels; infectious agents; early life exposures including prenatal factors; genetic influences; oestrogen deficiency; and the role of the psychosocial environment. None of these factors are as important as the established risk factors for epidemic CHD and their potential contribution for improving population health is limited or unclear. Research into unexplained variations in the occurrence of CHD and into life course influences and socioeconomic inequalities may provide extra leads to effective public health action. Especially important is research on the upstream social and economic determinants of CHD and its major risk factors, on the spread of the CHD epidemic to poorer populations, and into prevention policy and programme effectiveness. Available evidence supports the feasibility and effectiveness of population-wide prevention directed towards increasing the proportion of people at low risk of CHD. The vast majority of the public health effort should be directed to this approach rather than to the high risk individual approach. There is still a major gap between knowledge and action in preventing the CHD epidemics. PMID- 12540705 TI - Commentary: occupational therapy or the major challenge? PMID- 12540706 TI - Commentary: the epidemiology of self-deprecation. PMID- 12540707 TI - Commentary: using research evidence to promote cardiovascular health in children. PMID- 12540708 TI - Commentary: lifelong prevention of atherosclerosis: the critical importance of major risk factor exposures. PMID- 12540710 TI - Brothers who when the sirens roar. PMID- 12540712 TI - A simple statistical method for measuring how life events affect happiness. AB - BACKGROUND: Life events-like illness, marriage, or unemployment-have important effects on people. But there is no accepted way to measure the different sizes of these events upon human happiness and psychological health. By using happiness regression equations, economists have recently developed a method. METHODS: We estimate happiness regressions using large random samples of individuals. The relative coefficients of income and life events on happiness allow us to calculate a monetary 'compensating amount' for each kind of life event. RESULTS: The paper calculates the impact of different life events upon human well-being. Getting married, for instance, is calculated to bring each year the same amount of happiness, on average, as having an extra pound 70 000 of income per annum. The psychological costs of losing a job greatly exceed those from the pure drop in income. Health is hugely important to happiness. Widowhood brings a degree of unhappiness that would take, on average, an extra pound 170 000 per annum to offset. Well-being regressions also allow us to assess one of the oldest conjectures in social science-that well-being depends not just on absolute things but inherently on comparisons with other people. We find evidence for comparison effects. CONCLUSION: We believe that the new statistical method has many applications. In principle, it can be used to value any kind of event in life. PMID- 12540714 TI - A prospective study of job strain and coronary heart disease in US women. AB - BACKGROUND: Previous studies of job strain and coronary heart disease (CHD) in men have established job strain as a predictor of CHD risk. Despite the wealth of convincing evidence in men for an association between job strain and CHD, data in women remain sparse. METHODS: We prospectively evaluated the relation between job strain and CHD risk in the Nurses' Health Study. In this analysis, we followed a sample of 35 038 US female nurses aged 46-71 years, who completed questions about job strain in 1992 and who were free of diagnosed CHD, stroke, and cancer at baseline. The main outcome measure was the incidence of CHD occurring between baseline (1 June 1992) and 31 May 1996. RESULTS: During 4 years of follow-up, we documented 146 incident cases of CHD (108 non-fatal cases of myocardial infarction and 38 CHD deaths). No evidence was found for a relationship between job strain and risk of CHD. In multivariate analyses controlling for age, smoking, alcohol intake, body mass index, history of hypertension, diabetes mellitus, and other covariates, women in high strain jobs did not have an increased risk of CHD (relative risk [RR] = 0.71, 95% CI: 0.42-1.19) compared with women in low strain jobs. Neither women in passive jobs (RR = 1.08, 95% CI: 0.69-1.68) nor those in active jobs (RR = 0.91, 95% CI: 0.54-1.53) had an increased risk of CHD. CONCLUSIONS: Job strain was not related to an increase in the incidence of CHD in the present cohort of nurses. PMID- 12540716 TI - Serious health events following involuntary job loss in New Zealand meat processing workers. AB - BACKGROUND: The association between unemployment and poor health outcomes is well documented. Significant debate exists as to whether unemployment causes ill health or whether those with poor health find it harder to obtain and maintain employment. Factory closure studies are well placed to comment on causation. The objective of this study was to investigate associations between involuntary job loss, mortality and serious illness. METHODS: An 8-year follow-up of workers from two meat-processing plants in the Hawkes Bay region of New Zealand. A cohort (n = 1945) made redundant in 1986 was compared with a cohort (n = 1767) from the neighbouring plant that remained open until 1994. Incidence rates for mortality, cancer registrations and admissions to public hospitals were derived from record linkage with routinely collected national data. RESULTS: Follow-up for the period 1986-1994 was 96% complete for both plants. Among the cohort made redundant in 1986, there was an increased risk of serious self-harm which led to hospitalization or death (adjusted for age, sex and ethnicity relative risk [RR] = 2.47; 95% CI: 1.04-5.89) compared to the employed cohort. The RR of admission to hospital with a mental health diagnosis was 1.17 (95% CI: 0.68-2.01). There were no other statistically significant findings. CONCLUSIONS: This study has most of the features of an 'ideal' factory closure study, in that it achieved near-complete follow-up of a large workforce made redundant and a similar employed workforce for 8 years. We found that exposure to involuntary job loss increased the risk of mental distress leading to serious self-harm. No other association was found. PMID- 12540717 TI - Self-assessed health and mortality: could psychosocial factors explain the association? AB - BACKGROUND: The single-item question of self-assessed health has consistently been reported to be associated with mortality, even after controlling for a wide range of health measurements and known risk factors for mortality. It has been suggested that this association is due to psychosocial factors which are both related to self-assessed health and to mortality. We tested this hypothesis. METHODS: The study was carried out in a subsample (n = 5667) of the GLOBE population, a prospective cohort study conducted in the southeastern part of the Netherlands. Data on self-assessed health, sociodemographic variables, various aspects of health status, behavioural risk factors, and a number of psychosocial factors (social support, psychosocial stressors, personality traits, and coping styles) were collected by postal survey and structured interview in 1991, and mortality data were collected between 1991 and 1998. Cox proportional hazards analyses were used to calculate the association between self-assessed health and mortality, before and after controlling for the psychosocial variables. RESULTS: After controlling for sociodemographic variables, various aspects of health status, and behavioural risk factors, self-assessed health is still strongly associated with mortality in our dataset (Relative Risk [RR] of dying for 'poor' versus 'very good' self-assessed health = 3.98; 95% CI: 1.65-9.61). After controlling for the same set of confounders, many of the psychosocial variables are statistically significantly associated with a 'less-than-good' self-assessed health, particularly instrumental social support, long-lasting difficulties, neuroticism, and locus of control. However, only 'disclosure of emotions'-coping style has a statistically significant relationship with mortality. Adding the psychosocial variables to a model already containing self-assessed health does not attenuate the association between self-assessed health and mortality. CONCLUSIONS: We did not find indications that the association between self assessed health and mortality is due to the psychosocial factors included in this analysis. It seems likely that the unexplained mortality effects of self-assessed health are due to the fact that self-assessed health is a very inclusive measure of health reflecting health aspects relevant to survival which are not covered by other health indicators. PMID- 12540718 TI - Work-related psychosocial factors and carotid atherosclerosis. AB - BACKGROUND: In order to better understand the role of work environment in the earlier stages of the cardiovascular disease process, we wanted to investigate the influence of work-related psychosocial factors on preclinical atherosclerosis. METHODS: Cross-sectional data was used to examine the association between psychological job demands, job decision latitude, and carotid atherosclerosis in 2658 vocationally-active Swedish men and women, ages 46-65, from the general population. Odds ratios of carotid plaque prevalence and carotid artery intima-media thickness (IMT), determined by B-mode ultrasound, were estimated across combinations of job demands and decision latitude. RESULTS: Women in job situations with high demands and low decision latitude ('job strain') showed a high plaque prevalence odds (odds ratio [OR] = 1.68, 95% CI: 1.14, 2.48), and a thicker IMT in the carotid bifurcation area (mean difference: 0.15 mm, 95% CI: 0.07, 0.23) compared with women in job situations with low demands and high decision latitude ('relaxed'). Adjustment for covariates only slightly reduced the magnitude of these associations. No such associations were seen in men. However, women in job situations with high demands and high decision latitude ('active') also showed high odds for carotid plaque, and a thicker IMT in the carotid bifurcation, compared with women in 'relaxed' job situations. In men, those in 'active' job situations had a low carotid plaque prevalence odds, while IMT in the carotid bifurcation did not differ from those in 'relaxed' job situations. Results showed only weak associations with IMT in the common carotid artery (CCA) in both men and women. CONCLUSION: The specific hypothesis that high job demands interact synergistically with low decision latitude in the development of carotid atherosclerosis could not be supported in this study, neither in men nor in women. Instead a more complex pattern of interaction between job demands and decision latitude was shown. PMID- 12540719 TI - In-hospital symptoms of depression do not predict mortality 3 years after myocardial infarction. AB - BACKGROUND: The main aim of this study was to examine the relationship between symptoms of depression following myocardial infarction (MI) and 3-year survival status. METHODS: The Beck Depression Inventory was completed by 288 patients hospitalized for MI. Patients' cardiological status, including indices of disease severity, were recorded or derived from hospital notes. Three-year survival status was determined using patient information systems and cause of death ascertained from death certificates. RESULTS: During the 3 years of follow-up, 38 patients (13%) died, 33 (11%) from cardiac causes. Symptoms of depression did not predict either cardiac-specific or all-cause mortality. Similarly, in-hospital levels of anxiety were not associated with prognosis. In contrast, measures of disease severity and discharge medication status were strong prognostic indicators. Depression was not related to measures of disease severity at entry to the study. CONCLUSIONS: Symptoms of depression following MI do not predict longer-term survival, although measures of disease severity and discharge medication status do. Previous positive results for depression and cardiac mortality in MI patients could reflect the occasional confounding of depression with disease severity. PMID- 12540720 TI - Social determinants of disability pension: a 10-year follow-up of 62 000 people in a Norwegian county population. AB - BACKGROUND: Non-medical factors may be important determinants for granting disability pension (DP) even though disability is medically defined, as in Norway. The aim of this analysis was to identify determinants of DP in a total county population in a 10-year follow-up study. METHODS: Participants were people without DP, 20- to 66-years-old in 1984-1986. The baseline data were obtained in the Nord-Trondelag Health Study (HUNT): 90 000 people were invited to answer questionnaires on health, disease, social, psychological, occupational, and lifestyle factors. Information on those who later received DP was obtained from the National Insurance Administration database in 1995. Data analyses were performed using Cox regression analyses. RESULTS: The incidence of DP showed great variation with regards to age and gender, accounting for an overall increase in the follow-up period. Low level of education, low self-perceived health, occupation-related factors and any long-standing health problem were found to be the strongest independent determinants of DP. Low level of education and socioeconomic factors contributed more to younger people's risk compared to those over 50 years. For people under 50 years of age with a low level of education compared to those with a high level of education, the age-adjusted relative risk for DP was 6.35 for men and 6.95 for women. The multivariate adjusted relative risk was 2.91 and 4.77, respectively. CONCLUSIONS: Even for a medically based DP, low socioeconomic status, low level of education and occupational factors might be strong determinants when compared to medical factors alone. These non-medical determinants are usually not addressed by individual based health or rehabilitation programmes. PMID- 12540721 TI - Multiple measures of socio-economic position and psychosocial health: proximal and distal measures. AB - BACKGROUND: The aim of this paper is to compare three models for exploring the links between different measures of adult socioeconomic position (SEP)-education, occupation, income-and psychosocial health. Model I is a basic univariate regression model with psychosocial health as the outcome and a measure of SEP as the predictor. Model II is a multiple regression model with psychosocial health as the outcome with all three measures of SEP allocated the same temporal position as predictors. Model III treats education, a distal measure of SEP, as antecedent to the proximal measures of SEP in the prediction equations linking SEP to health. METHODS: Participants were drawn from the Whitehall II study, a prospective cohort study of British civil servants. Data analysed here are from Phase 5 (1997-1999) of data collection, 7830 individuals in all. The measures of SEP and psychosocial health were assessed via a self-administered questionnaire. RESULTS: The three models can lead to completely different conclusions. Model III, our preferred model, shows education to have a stronger indirect effect on psychosocial health when compared to its direct effect. The indirect effect is due to the effect of education on proximal measures of social position, occupation, and income in this case. CONCLUSIONS: Results reported here support the hypothesis that a comparison of the relative importance of the different measures of social position in predicting health is meaningless if the causal relationships among these measures are not accounted for. PMID- 12540723 TI - Risk factors for depression in postnatal first year, in eastern Turkey. AB - BACKGROUND: There are few studies reporting depression in the postnatal period in developing countries. In this study our objective was to evaluate women from eastern Turkey in the postnatal one-year period in order to analyse the risk factors for depression. METHODS: In this cross-sectional, multi-centre study, we selected a study sample from five eastern provinces. Among 2602 randomly selected women who gave birth within the last year, we included 2514 women in our analysis. The Edinburgh Postnatal Depression Scale was used for the evaluation of depression. RESULTS: The percentage of women with high depression scores was 27.2%. Excess risk of depression was associated with several factors including unemployment, low education, poverty, poor family relations, low marital age, lack of medical services, and mental health problems. CONCLUSIONS: Depression in postnatal women is an important public health problem in the less developed eastern part of Turkey. PMID- 12540724 TI - Social networks and mortality based on the Komo-Ise cohort study in Japan. AB - BACKGROUND: No prospective studies have examined the association between social networks and all-cause and cause-specific mortality among middle-aged Japanese. The study of varied populations may contribute to clarifying the robustness of the observed effects of social networks and extend their generalizability. METHODS: To clarify the association between social networks and mortality among middle-aged and elderly Japanese, a community-based prospective study, the Komo Ise Study, was conducted in two areas of Gunma Prefecture, Japan. A total of 11 565 subjects aged 40-69 years at baseline in 1993 completed a self-administered questionnaire. During the 7-year follow-up period, 335 men and 155 women died and the relative risk (RR) of each social network item was estimated by the Cox proportional hazard model. RESULTS: Single women had significantly increased risks of all-cause (multivariate RR = 2.2), and all circulatory system disease (age-area adjusted RR = 2.6) mortality. Men who did not participate in hobbies, club activities, or community groups had significantly higher multivariate RR for all-cause (RR = 1.5), all circulatory system disease (RR = 1.6) and non-cancer and non-circulatory system disease (RR = 2.3) mortality. Urban women who rarely or never met close relatives had significantly elevated risks of all-cause (RR = 2.4), all cancer (RR = 2.6), and non-cancer and non-circulatory system disease (RR = 2.7) mortality after adjustment for established risk factors. CONCLUSIONS: This study provides evidence that social networks are an important predictor of mortality risk for middle-aged and elderly Japanese men and women. Lack of participation, for men, and being single and lack of meeting close relatives, for women, were independent risk factors for mortality. PMID- 12540725 TI - Psychosocial risk factors for the onset of abdominal pain. Results from a large prospective population-based study. AB - OBJECTIVE: To determine the psychosocial risk factors for the development of abdominal pain and to determine whether, in those people who consulted, symptoms had been attributed to an organic cause. DESIGN: Prospective population-based postal survey with follow-up survey at 12 months. SETTING: A mixed sociodemographic suburban area of Manchester, UK. PARTICIPANTS: Subjects aged 18 65 years were randomly selected from a population-based primary care register who had responded to a detailed pain questionnaire, which included a pain manikin drawing. They also completed the following psychosocial instruments: General Health Questionnaire, Somatic Symptom Checklist, Fatigue Questionnaire and the Illness Attitude Scales (including the 'health anxiety' and 'illness behaviour' sub-scales). MAIN OUTCOME MEASURES: The onset of new abdominal pain. RESULTS: Of the 1953 participants at baseline, 1763 were free of abdominal pain: 1551 were followed up at 12 months (adjusted follow-up rate of 92%) of which 69 subjects reported new abdominal pain (new onset rate 4.6%). New abdominal pain was similar in females (4.9%) and males (4.2%), and did not vary by age group. Baseline factors which predicted onset were high levels of fatigue (odds ratio [OR] = 3.3, 95% CI: 1.9-5.8), psychological distress (OR = 3.4, 95% CI: 1.9-6.0), high scores on the illness behaviour scale (OR = 3.3, 95% CI: 1.7-6.7) and high levels of health anxiety (OR = 2.1, 95% CI: 1.1-3.9). Reporting low back pain at baseline was also associated with an increased risk of reporting abdominal pain (OR = 2.0, 95% CI: 1.2-3.3). On multivariate analysis, high levels of psychological distress and aspects of prior illness behaviour were the major independent predictors of outcome. Of those who sought health care, only one consultation led to a definite diagnosis. CONCLUSION: In subjects free of abdominal pain, psychological distress, fatigue, health anxiety and illness behaviour are predictors of future onset rather than merely a consequence of symptoms. These results suggest that abdominal pain shares some common features of onset with pain at other sites thought not to be primarily organic in origin. PMID- 12540728 TI - Fetal origins of adult disease: strength of effects and biological basis. AB - BACKGROUND: Low birthweight has been consistently shown to be associated with coronary heart disease (CHD) and its biological risk factors. The effects of low birthweight are increased by slow infant growth and rapid weight gain in childhood. To quantify the importance of developmental processes in the genesis of CHD it is necessary to establish the impact of fetal, infant and childhood growth on major pathological events in later life-death, hospital treatment and the need for medication. METHODS: Longitudinal study of 13 517 men and women who were born in Helsinki University Hospital during 1924-1944, whose body sizes at birth and during childhood were recorded, and in whom deaths, hospital admissions, and prescription of medication for chronic disease are documented. RESULTS: The combination of small size at birth and during infancy, followed by accelerated weight gain from age 3 to 11 years, predicts large differences in the cumulative incidence of CHD, type 2 diabetes and hypertension. CONCLUSIONS: Coronary heart disease and type 2 diabetes may originate through two widespread biological phenomena-developmental plasticity and compensatory growth. PMID- 12540727 TI - Psychosocial risk factors for heart disease in France and Northern Ireland: the Prospective Epidemiological Study of Myocardial Infarction (PRIME). AB - BACKGROUND: France has a substantially lower level of premature mortality from cardiovascular diseases (CVD) relative to its comparators. Compared with Northern Ireland, France has one-half the rate, despite having a similar cardiovascular risk profile to Northern Ireland. In this prospective longitudinal study the psychosocial risk hypothesis for CVD was tested. METHOD: A cohort of 9758 men (7359 in France and 2399 in Northern Ireland) aged 50-59 years who were initially free of any CVD were recruited. At baseline the subjects completed a psychosocial questionnaire, measuring hostility, depression, social support, and the Type A behaviour pattern. At 5-years follow-up their clinical status was determined. RESULTS: Multivariate analysis indicated that, contrary to prediction, France had a substantially more negative psychosocial risk profile than Northern Ireland. The psychosocial risk factors were not successful at predicting at 5-years follow up the hard clinical endpoint of definite fatal/non-fatal myocardial infarction. In the case of the softer clinical endpoint, angina pectoris/unstable angina, only depression predicted outcome with a small effect size. CONCLUSION: The findings provide little support for the psychosocial risk hypothesis. The psychosocial risk profile was more negative in France, the opposite of that predicted. The finding of a relationship between depression and angina may reflect a tendency for individuals who respond negatively on mood state to report more cardiac symptoms irrespective of physical disease state. PMID- 12540729 TI - Fetal growth and hospitalization with asthma during early childhood: a follow-up study in Denmark. AB - BACKGROUND: Childhood asthma may have a fetal origin. In order to examine this hypothesis we examined the association between fetal growth indicators and hospitalization with asthma during early childhood. METHODS: We performed a cohort study with 10 440 children whose mothers attended the midwife centres in Odense and Aalborg, Denmark, from April 1984 to April 1987 at approximately the 36th week of gestation. Information on hospitalization with asthma was extracted from the National Hospital Discharge Registry, which allowed a complete follow-up of all members of the cohort who remained alive and residents in the county. RESULTS: The cumulative incidence rates of hospitalization with 'definite' and 'any' asthma were 3.5% and 5.0%, respectively, at 12 years of age. After adjusting for potential confounders, there were increased trends in the risk of asthma hospitalization with birthweight and ponderal index. More pronounced trends, particularly in the risk of definite asthma, were found among children of over the average birthweight (IRR = 1.62, 95% CI: 1.02-2.59 per 1000 g increase) or ponderal index (IRR = 1.06, 95% CI: 1.02-1.10 per 0.1 g/cm(3) increase). However, the increased risk in large babies was seen only in children with a ponderal index of > or =2.5 g/cm3. CONCLUSION: Birthweight and ponderal index were associated with childhood asthma indicated by hospitalization. Fetal growth itself, determinants of fetal growth, or any unadjusted factors that correlate with fetal growth, could cause the association. PMID- 12540730 TI - Probabilistic record linkage and a method to calculate the positive predictive value. AB - BACKGROUND: Computerized record linkage is commonly used in cohort studies to ascertain the study outcome, and as such its accuracy classifying the outcome can be described using the standard epidemiological terms of sensitivity and positive predictive value (PPV). METHOD: We describe a 'duplicate method' to calculate the PPV of record linkage when each record can only be involved in one match (e.g. linking population files to death files). The method does not require a validation subset of records from both files with detailed personal information (e.g. name and address), and is therefore ideal for linkage projects using anonymous data. The duplicate method assumes that the number of records from one file with zero, one, two, etc., links from the other file is distributed in a manner predicted by combinatorial probabilities. Having made this assumption, the number of false positive links, and hence the PPV, are estimable. We demonstrate this duplicate method using output from anonymous and probabilistic record linkage of census and mortality records in New Zealand. RESULTS: The PPV estimates conform to the pattern expected based on the underlying theory of probabilistic record linkage, and were robust to sensitivity analyses. We encourage other researchers to further assess the accuracy of this method. PMID- 12540731 TI - Prediction of risk of coronary events in middle-aged men in the Prospective Cardiovascular Munster Study (PROCAM) using neural networks. AB - BACKGROUND: Logistic regression (LR) is commonly used to estimate risk of coronary heart disease. We investigated if neural networks improved on the risk estimate of LR by analysing data from the Prospective Cardiovascular Munster Study (PROCAM), a large prospective epidemiological study of risk factors for coronary heart disease among men and women at work in northern Germany. METHODS: We used a multi-layer perceptron (MLP) and probabilistic neural networks (PNN) to estimate the risk of myocardial infarction or acute coronary death (coronary events) during 10 years' follow-up among 5159 men aged 35-65 years at recruitment into PROCAM. In all, 325 coronary events occurred in this group. We assessed the performance of each procedure by measuring the area under the receiver-operating characteristics curve (AUROC). RESULTS: The AUROC of the MLP was greater than that of the PNN (0.897 versus 0.872), and both exceeded the AUROC for LR of 0.840. If 'high risk' is defined as an event risk >20% in 10 years, LR classified 8.4% of men as high risk, 36.7% of whom suffered an event in 10 years (45.8% of all events). The MLP classified 7.9% as high risk, 64.0% of whom suffered an event (74.5% of all events), while with the PNN, only 3.9% were at high risk, 58.6% of whom suffered an event (33.5% of all events). CONCLUSION: Intervention trials indicate that about one in three coronary events can be prevented by 5 years of lipid-lowering treatment. Our analysis suggests that use of the MLP to identify high-risk individuals as candidates for drug treatment would allow prevention of 25% of coronary events in middle-aged men, compared to 15% and 11% with LR and the PNN, respectively. PMID- 12540734 TI - Homocysteine and coronary heart disease: the importance of a distinction between low and high risk subjects. PMID- 12540735 TI - Cold temperature and consultations for respiratory and cardiovascular disease. PMID- 12540736 TI - The case for an epidemiology of terrorism. PMID- 12540738 TI - Toward a consensus on 'race' and birthweight distributions. PMID- 12540737 TI - Olive oil consumption and risk of non-fatal myocardial infarction in Italy. PMID- 12540740 TI - Induction of drug-metabolizing enzymes: a path to the discovery of multiple cytochromes P450. AB - This article provides a personal account of the discovery of the induced synthesis of drug-metabolizing enzymes and of subsequent research that led to the discovery of multiple cytochromes P450 with different catalytic activities. The manuscript also emphasizes the role of environmental factors (in addition to genetic polymorphisms) in explaining person-to-person and day-to-day differences in rates and pathways of drug metabolism that occur in the human population. PMID- 12540741 TI - Novel pharmacological approaches to manage interstitial lung fibrosis in the twenty-first century. AB - Pharmacological agents currently in use to treat interstitial lung fibrosis are either ineffective or too toxic in humans. This review addresses mechanistically based novel approaches that have the potential to minimize the accumulation of collagen in the lung, a hallmark of lung fibrosis. These approaches include maintaining the intracellular levels of NAD(+) and ATP, blocking the biological activities of TGF-beta and integrins, evaluating the effectiveness of PAF receptor antagonists and NOS inhibitors, and developing a new generation of cysteine pro-drugs with an adequate degree of bioavailability. A critical analysis of each approach as it relates to management of IPF in humans is presented. PMID- 12540742 TI - Nitric oxide-releasing drugs. AB - Pharmacological compounds that release nitric oxide (NO) have been useful tools for evaluating the broad role of NO in physiology and therapeutics. NO deficiency has been implicated in the genesis and evolution of several disease states. Both medical needs and commercial opportunities have fostered attempts to modulate NO in the human body for therapeutic gain. Strategies for NO modulation encompass antiinflammatory, sexual dysfunction, and cardiovascular indications. Apart from newly developed drugs, several commonly used cardiovascular drugs exert their beneficial action, at least in part, by modulating the NO pathway. This review discusses the fundamental pharmacological properties and mechanisms of action of NO-releasing drugs. Some of these compounds may enter in the clinical arena providing important therapeutic benefits in human diseases. PMID- 12540743 TI - Activation of the aryl hydrocarbon receptor by structurally diverse exogenous and endogenous chemicals. AB - The induction of expression of genes for xenobiotic metabolizing enzymes in response to chemical insult is an adaptive response found in most organisms. In vertebrates, the AhR is one of several chemical/ligand-dependent intracellular receptors that can stimulate gene transcription in response to xenobiotics. The ability of the AhR to bind and be activated by a range of structurally divergent chemicals suggests that the AhR contains a rather promiscuous ligand binding site. In addition to synthetic and environmental chemicals, numerous naturally occurring dietary and endogenous AhR ligands have also been identified. In this review, we describe evidence for the structural promiscuity of AhR ligand binding and discuss the current state of knowledge with regards to the activation of the AhR signaling pathway by naturally occurring exogenous and endogenous ligands. PMID- 12540744 TI - Trafficking of NMDA receptors. AB - The NMDA receptor (NMDAR) plays a central role in the function of excitatory synapses. Recent studies have provided interesting insights into several aspects of the trafficking of this receptor in neurons. The NMDAR is not a static resident of the synapse. Rather, the number and composition of synaptic NMDARs can be modulated by several factors. The interaction of PDZ proteins, generally thought to occur at the synapse, appears to occur early in the secretory pathway; this interaction may play a role in the assembly of the receptor complex and its exit from the endoplasmic reticulum. This review addresses recent advances in our understanding of NMDAR trafficking and its synaptic delivery and maintenance. PMID- 12540745 TI - Telomere inhibition and telomere disruption as processes for drug targeting. AB - The components and cofactors of the holoenzyme telomerase and its substrate telomeric DNA are attractive targets for anticancer agents that act by inhibiting the activity of telomerase. This review outlines recent advances in telomerase inhibition that have been achieved using antisense oligonucleotides and ribozymes that target the telomerase mRNA or its hTR RNA template. Although these are potent catalytic inhibitors of telomerase, they are challenging to implement in the clinic due to their delayed effectiveness. Drugs that directly bind to the telomeres, the complex structures that are associated at the telomeric ends, and stabilize secondary DNA structures such as G-quadruplexes are also potent inhibitors of telomerase. Special focus is given here to the telomeres, the biological machinery that works in tandem with telomerase to elongate telomeres, the causes of telomere disruption or dysfunction, and the consequences of disruption/dysfunction on the activity and design of anticancer agents. PMID- 12540746 TI - Pharmacology and physiology of human adrenergic receptor polymorphisms. AB - Adrenergic receptors are expressed on virtually every cell type in the body and are the receptors for epinephrine and norepinephrine within the sympathetic nervous system. They serve critical roles in maintaining homeostasis in normal physiologic settings as well as pathologic states. These receptors are also targets for therapeutically administered agonists and antagonists. Recent studies have shown that at least seven adrenergic receptor subtypes display variation in amino acid sequence in the human population due to common genetic polymorphisms. Variations in potential regulatory domains in noncoding sequence are also present. Here, we review the consequences of these polymorphisms in terms of signaling, human physiology and disease, and response to therapy. PMID- 12540747 TI - K+ channel structure-activity relationships and mechanisms of drug-induced QT prolongation. AB - Pharmacological intervention, often for the purpose of treating syndromes unrelated to cardiac disease, can increase the vulnerability of some patients to life-threatening rhythm disturbances. This may be due to an underlying propensity stemming from genetic defects or polymorphisms, or structural abnormalities that provide a substrate allowing for the initiation of arrhythmic triggers. A number of pharmacological agents that have proven useful in the treatment of allergic reactions, gastrointestinal disorders, and psychotic disorders, among others, have been shown to reduce repolarizing K(+) currents and prolong the QT interval on the electrocardiogram. Understanding the structural determinants of K(+) channel blockade may provide new insights into the mechanism and rate-dependent effects of drugs on cellular physiology. Drug-induced disruption of cellular repolarization underlies electrocardiographic abnormalities that are diagnostic indicators of arrhythmia susceptibility. PMID- 12540748 TI - Complementary and alternative therapeutics: rigorous research is needed to support claims. AB - The establishment of the National Center for Complementary and Alternative Medicine (NCCAM) in 1998 as a part of the National Institutes of Health was catalyzed by the increasing interest and use of complementary and alternative medicine (CAM) modalities by the public. This article presents an overview of CAM, summarizes similarities and differences between the regulatory requirements for drugs and CAM/botanical products, identifies several challenges and opportunities for conducting research to demonstrate the safety and efficacy of CAM therapeutics, and highlights the role of NCCAM in supporting and stimulating research in this area. PMID- 12540749 TI - Innate immune responses to microbial poisons: discovery and function of the Toll like receptors. AB - There are many circumstances under which a toxin exploits an endogenous receptor or another protein of host origin to work its untoward effects. In most instances, the receptor normally fulfills a function that has nothing to do with the toxin per se; that is, the toxin is not the "natural" ligand. The situation with endotoxin, however, is a remarkable one. The endotoxin receptor evolved to detect endotoxin. Why have mammals maintained a gene that can undermine their survival? The search for the endotoxin receptor answered this question and also revealed the essential function and biological strategy of the Toll-like receptors: principal sensors of the innate immune system. PMID- 12540753 TI - [Image display devices (1)]. PMID- 12540754 TI - [Run through the IHE technical framework]. PMID- 12540755 TI - [Current state of breast tumors imaging]. PMID- 12540756 TI - [Medical imaging technology of the pancreatico-biliary systems: special emphasis on MR imaging in the era of multislice CT]. PMID- 12540757 TI - [Biological effects of low dose radiation and adaptive responses in mammalian cells]. PMID- 12540758 TI - [Neuroimaging of Alzheimer disease and vascular dementia]. PMID- 12540759 TI - [Dosimetry in radiotherapy]. PMID- 12540760 TI - [Apparatus engineering of X ray CT equipment (9) : dose evaluation]. PMID- 12540763 TI - [Research and studies on design and production for medical diagnostic imaging systems considered environmental problems part 1]. PMID- 12540764 TI - [Fundamental study of DSA images using gadolinium contrast agent]. AB - Most contrast agents used in digital subtraction angiography (DSA) are non-ionic iodinated contrast agents, which can cause severe side effects in patients with contraindications for iodine or allergic reactions to iodine. Therefore, DSA examinations using carbon dioxide gas or examinations done by magnetic resonance imaging (MRI) and ultrasound (US) were carried out in these patients. However, none of these examinations provided images as clear as those of DSA with an iodinated contrast agent. We experienced DSA examination using a gadolinium contrast agent in a patient contraindicated for iodine. The patient had undergone MRI examination with a gadolinium contrast agent previously without side effects. The characteristics of gadolinium and the iodinated contrast agent were compared, and the DSA images obtained clinically using these media were also evaluated. The signal-to-noise (SN) ratio of the gadolinium contrast agent was the highest at tube voltages of 70 to 80 kilovolts and improved slightly when the image intensifier (I.I.) entrance dose was greater than 300 microR (77.4 nC/kg). The dilution ratios of five iodinated contrast agents showed the same S/N value as the undiluted gadolinium contrast agent. Clinically, the images obtained showed a slight decrease in contrast but provided the data necessary to make a diagnosis and made it possible to obtain IVR without any side effects. DSA examinations using a gadolinium contrast agent have some benefit with low risk and are thought to be useful for patients contraindicated for iodine. PMID- 12540765 TI - [Study of gradient echo (GRE) Dixon image using low-field MRI scanner for examination of metastatic bone marrow tumors: comparison of double and single echo sequences]. AB - We evaluated the gradient echo (GRE) Dixon method in metastatic bone tumors using a low-field MRI scanner (0.2 Tesla). This method is characterized by the double echo sequence of in-phase and opposed-phase. Studies were carried out on a phantom, 14 healthy volunteers, and clinical examples (33 vertebral bodies) using the T(1)-weighted spin echo, T(2)-weighted turbo spin echo, and GRE Dixon methods. Further, we obtained addition and subtraction images from the double echo sequence. In the clinical examples, the contrast-to-noise ratio (CNR) of the subtraction images (51.3+/-24.1) was significantly better than that of the T(1) SE images (6.7+/-3.1, p<0.0001). For the examination of metastatic bone marrow tumors using a low-field MRI scanner (0.2 Tesla), subtraction images are thought to be the most effective. PMID- 12540766 TI - [Visual impression of photographic density of the brain in head CT images: verification of optical illusion by visual subjective evaluation of medical images]. AB - The purpose of this study was to clarify the psychological influence of optical illusion on the photographic density of the brain in head CT images. In this study, I attempted to compare the visual impression of photographic density of the brain using visual subjective evaluation according to observers and cases. The results showed that the visual impression of photographic density of the brain at cerebellar CT depended upon the mode of appearance of the optical illusion (assimilation, contrast, picture frame effect, etc.). In addition, that impression varied according to differences in cases (difference of picture pattern) and observers. Further, it was clarified that the optical illusion had been observed according to individual differences among observers. Moreover, cases could be divided into two main classes. Some cases were recognized as a kind of outline figure (the white skull was recognized as an induction color, and the brain as a non-induction color) with assimilation, and consequently the brain seemed whitish. In other cases (the white skull was not recognized as an induction color) in which there was no assimilation, the brain did not seem whitish. This was considered to have occurred because the degree of influence of the skull on the visual impression of the photographic density of the brain had changed according to differences in cases. PMID- 12540767 TI - [Medical image transfer for emergency care utilizing internet and mobile phone]. AB - The purpose of this study was to utilize a mobile phone for transferring medical images to a specialist at a remote site in emergency care. CT images obtained in emergency treatment are transmitted via the Internet or mobile phone (CDMA: Code Division Multiple Access communication method). A specialist even at a remote mobile site may receive the appended images of e-mail sent via mobile phone to his PDA (Personal Digital Assistant, image indicator). The images are originally obtained in DICOM format by CT, and are then converted to JPEG format. To reduce transmission time, the JPEG files can be compressed without causing undue deterioration in image quality. To evaluate the quality of images received at the remote site, images were compressed at 25, 40, 50, 60, 70 and 80% compression ratios and transmitted to a PDA via the Internet and a mobile phone or a mobile phone alone. Image quality was visually evaluated by BVC analysis. The analysis revealed that the image compression ratio must be at least 50% for diagnosis. Considering the transmission speed, a 50% compression ratio was considered the most suitable for this application. PMID- 12540768 TI - [Development and evaluation of QA software for the monitor chamber of an accelerator]. AB - The monitor chamber of a radiotherapy system needs to be calibrated once a week. Because the calibration procedure requires a large variety of complicated calculations, we have developed software that facilitates calculation and enables comparison and storage of data. According to the standard measurement of absorbed dose, we used Visual Basic 6.0 (Microsoft Corp.) to establish the calibration method. This new technique has simplified the conventional intricate calculation required for calibration of the monitor chamber and enabled automatic processing of calculated results. We have confirmed the usefulness of this software in calibrating the monitor chamber. In the routine inspection, we can compare the current data with former results. Because of this advantage, it is possible to eliminate serious accidents such as overdosing and underdosing. PMID- 12540770 TI - Acute renal failure in patients with cirrhosis: perspectives in the age of MELD. AB - In patients with cirrhosis, acute renal failure is mainly due to prerenal failure (caused by renal hypoperfusion) and tubular necrosis. The main causes of prerenal failure are "true hypovolemia" (induced by hemorrhage or gastrointestinal or renal fluid losses), sepsis, or type 1 hepatorenal syndrome (HRS). The frequency of prerenal failure due to the administration of nonsteroidal anti-inflammatory drugs or intravascular radiocontrast agents is unknown. Prerenal failure is rapidly reversible after restoration of renal blood flow. Treatment is directed to the cause of hypoperfusion, and fluid replacement is used to treat most cases of "non-HRS" prerenal failure. In patients with type 1 HRS with very low short term survival rate, liver transplantation is the ideal treatment. Systemic vasoconstrictor therapy (with terlipressin, noradrenaline, or midodrine [combined with octreotide]) may improve renal function in patients with type 1 HRS waiting for liver transplantation. MARS (for molecular adsorbent recirculating system) and the transjugular intrahepatic portosystemic shunt may also improve renal function in these patients. In patients with cirrhosis, acute tubular necrosis is mainly due to an ischemic insult to the renal tubules. The most common condition leading to ischemic acute tubular necrosis is severe and sustained prerenal failure. Little is known about the natural course and treatment (i.e., renal replacement therapy) of cirrhosis-associated acute tubular necrosis. PMID- 12540771 TI - Nonalcoholic steatohepatitis clinical research network. PMID- 12540772 TI - Oxidative and nitrosative stress in ammonia neurotoxicity. PMID- 12540773 TI - Empiric beta-blockers for the prophylaxis of variceal hemorrhage: cost effective or clinically applicable? PMID- 12540774 TI - Portopulmonary hypertension: new clinical insights and more questions on pathogenesis. PMID- 12540775 TI - Selective Na+/H+ exchange inhibition by cariporide reduces liver fibrosis in the rat. AB - The aim of this study was to evaluate the effect of cariporide, a selective Na(+)/H(+) exchange inhibitor, on isolated and cultured hepatic stellate cells (HSCs) and in 2 in vivo models of rat liver fibrosis. Platelet-derived growth factor (PDGF)-induced HSC proliferation, evaluated by measuring the percentage of bromodeoxyuridine-positive cells, was significantly inhibited by cariporide, with a maximal effect at 10 micromol/L. Incubation with cariporide did not inhibit PDGF-induced extracellular-regulated kinase 1/2 (ERK1/2), Akt (a downstream component of the phosphatidylinositol [PI]-3 kinase pathway), and protein kinase C (PKC) activation but reduced PDGF-induced activation of the Na(+)/H(+) exchanger, with a maximal effect at 10 micromol/L. Rats treated with dimethylnitrosamine (DMN; 10 mg/kg) for 1 and 5 weeks received a diet with or without 6 ppm cariporide. Treatment with cariporide reduced the degree of liver injury, as determined by alanine aminotransferase (ALT) values, also when administered after the induction of hepatic damage. This was associated with reduced HSC activation and proliferation and reduced collagen deposition, as determined by morphometric evaluation of alpha-smooth muscle actin (SMA)/proliferating cell nuclear antigen-positive cells and percentage of Sirius red-positive parenchyma, respectively. Moreover, cariporide was also able to reduce alpha(1)I procollagen messenger RNA (mRNA) expression. Similar effects were observed in bile duct-ligated (BDL) rats. In conclusion, selective inhibition of the Na(+)/H(+) exchanger by cariporide may represent an effective therapeutic strategy in the treatment of hepatic fibrosis. PMID- 12540776 TI - DNase I-hypersensitive sites enhance alpha1(I) collagen gene expression in hepatic stellate cells. AB - Liver fibrosis is characterized by a dramatic increase in the expression of type I collagen. Several deoxyribonuclease (DNase) I-hypersensitive sites (HS) have been located in the distal 5'-flanking region of the alpha1(I) collagen gene that are specific to collagen-producing cells. To assess the role of the DNase I-HS in regulating alpha1(I) collagen gene expression in hepatic stellate cells (HSCs), 3 transgenic mouse lines expressing collagen-alpha1(I) reporter genes were used (Krempen et al. Gene Expr 1999;8:151-163). The pCol9GFP transgene contains the collagen gene promoter (-3122 to +111) linked to the green fluorescent protein (GFP) reporter gene. The pCol9GFP-HS4,5 transgene contains HS4,5 and pColGFP HS8,9 contains HS8,9 positioned upstream of the collagen promoter in pCol9GFP. HSCs isolated from transgenic mice containing pCol9GFPHS4,5 and pColGFP-HS8,9 showed earlier and higher GFP expression patterns than HSCs isolated from pCol9GFP mice. HSCs from pCol9GFP-HS4,5 showed the highest levels of GFP expression and culture-induced expression correlated with induction of the endogenous alpha1(I) collagen gene. After CCl(4) administration, pCol9GFP-HS4,5 mice showed increased GFP expression compared with pCol9GFP mice in both whole liver extracts and isolated HSCs. Several sites for DNA-protein interactions in both HS4 and HS5 were identified that included a binding site for activator protein 1. In conclusion, DNase I-HS4,5 enhance expression of the alpha1(I) collagen gene promoter in HSCs both in vitro and in vivo after a fibrogenic stimulus. The collagen-GFP transgenic mice provide a convenient and reliable model system to investigate the molecular mechanisms controlling increased collagen expression during fibrosis. PMID- 12540777 TI - Mechanisms of hepatocyte protection against hypoxic injury by atrial natriuretic peptide. AB - Atrial natriuretic peptide (ANP) reduces ischemia and/or reperfusion damage in several organs, but the mechanisms involved are largely unknown. We used freshly isolated rat hepatocytes to investigate the mechanisms by which ANP enhances hepatocyte resistance to hypoxia. The addition of ANP (1 micromol/L) reduced the killing of hypoxic hepatocytes by interfering with intracellular Na(+) accumulation without ameliorating adenosine triphosphate (ATP) depletion and pH decrease caused by hypoxia. The effects of ANP were mimicked by 8-bromo-guanosine 3', 5'-cyclic monophosphate (cGMP) and were associated with the activation of cGMP-dependent kinase (cGK), suggesting the involvement of guanylate cyclase coupled natriuretic peptide receptor (NPR)-A/B ANP receptors. However, stimulating NPR-C receptor with des-(Gln(18), Ser(19),Gly(20),Leu(21),Gly(22)) ANP fragment 4-23 amide (C-ANP) also increased hepatocyte tolerance to hypoxia. C ANP protection did not involve cGK activation but was instead linked to the stimulation of protein kinase C (PKC)-delta through G(i) protein- and phospholipase C-mediated signals. PKC-delta activation was also observed in hepatocytes receiving ANP. The inhibition of phospholipase C or PKC by U73122 and chelerythrine, respectively, significantly reduced ANP cytoprotection, indicating that ANP interaction with NPR-C receptors also contributed to cytoprotection. In ANP-treated hepatocytes, the stimulation of both cGK and PKC-delta was coupled with dual phosphorylation of p38 mitogen-activated protein kinase (MAPK). The p38 MAPK inhibitor SB203580 abolished ANP protection by reverting p38 MAPK-mediated regulation of Na(+) influx by the Na(+)/H(+) exchanger. In conclusion, ANP recruits 2 independent signal pathways, one mediated by cGMP and cGK and the other associated with G(i) proteins, phospholipase C, and PKC-delta. Both cGK and PKC-delta further transduce ANP signals to p38 MAPK that, by maintaining Na(+) homeostasis, are responsible for ANP protection against hypoxic injury. PMID- 12540778 TI - Induction of cellular resistance against Kupffer cell-derived oxidant stress: a novel concept of hepatoprotection by ischemic preconditioning. AB - Ischemic preconditioning (IP) triggers protection of the liver from prolonged subsequent ischemia. However, the underlying protective mechanisms are largely unknown. We investigated whether and how IP protects the liver against reperfusion injury caused by Kupffer cell (KC)-derived oxidants. IP before 90 minutes of warm ischemia of rat livers in vivo significantly reduced serum alanine aminotransferase (AST) levels and leukocyte adherence to sinusoids and postsinusoidal venules during reperfusion. This protective effect was mimicked by postischemic intravenous infusion of glutathione (GSH), an antioxidative strategy against KC-derived H(2)O(2). Interestingly, no additional protection was achieved by infusion of GSH to preconditioned animals. These findings and several additional experiments strongly suggest IP mediated antioxidative effects: IP prevented oxidant cell injury in isolated perfused rat livers after selective KC activation by zymosan. Moreover, IP prevented cell injury and pertubations of the intracellular GSH/GSSG redox system caused by direct infusion of H(2)O(2) (0.5 mmol/L). IP-mediated resistance against H(2)O(2) could neither be blocked by the adenosine A2a antagonist DMPX nor mimicked by A2a agonist CGS21680. In contrast, H(2)O(2) resistance was abolished by the p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB203580, but induced when p38 MAPK was directly activated by anisomycin. In conclusion, we propose a novel concept of hepatoprotection by IP: protection of liver cells by enhancing their resistance against KC-derived H(2)O(2). Activation of p38 MAPK and preservation of the intracellular GSH/oxidized glutathione (GSSG) redox system, but not adenosine A2a receptor stimulation, seems to be pivotal for the development of H(2)O(2) resistance in preconditioned livers. PMID- 12540779 TI - Stat4 and Stat6 signaling in hepatic ischemia/reperfusion injury in mice: HO-1 dependence of Stat4 disruption-mediated cytoprotection. AB - Ischemia/reperfusion (I/R) injury remains an important problem in clinical organ transplantation. There is growing evidence that T lymphocytes, and activated CD4+ T cells in particular, play a key role in hepatic I/R injury. This study analyzes the role of signal transducer and activator of transcription 4 (Stat4) and Stat6 signaling in liver I/R injury. Using a partial lobar warm ischemia model, groups of wild-type (WT), T cell-deficient, Stat4-/Stat6-deficient knockout (KO) mice were assessed for the extent/severity of I/R injury. Ninety minutes of warm ischemia followed by 6 hours of reperfusion induced a fulminant liver failure in WT and Stat6 KO mice, as assessed by hepatocellular damage (serum alanine aminotransferase [sALT] levels), neutrophil accumulation (myeloperoxidase [MPO] activity) and histology (Suzuki scores). In contrast, T cell deficiency (nu/nu mice) or disruption of Stat4 signaling (Stat4 KO mice) reduced I/R insult. Unlike adoptive transfer of WT or Stat6-deficient T cells, infusion of Stat4-deficient T cells failed to restore hepatic I/R injury and prevented tumor necrosis factor alpha (TNF-alpha) production in nu/nu mice. Diminished TNF-alpha/Th1-type cytokine messenger RNA (mRNA)/protein elaborations patterns, along with overexpression of heme oxygenase-1 (HO-1)-accompanied hepatic cytoprotection in Stat4 KO recipients. In contrast, HO-1 depression restored hepatic injury in otherwise I/R resistant Stat4 KOs. In conclusion, Stat4 signaling is required for, whereas Stat4 disruption protects against, warm hepatic I/R injury in mice. The cytoprotection rendered by Stat4 disruption remains HO-1-dependent. PMID- 12540780 TI - Endogenous IL-13 protects hepatocytes and vascular endothelial cells during ischemia/reperfusion injury. AB - Hepatic ischemia/reperfusion injury involves a complex inflammatory cascade resulting in neutrophil-mediated injury of hepatocytes. Previous studies from our laboratory have established that exogenous administration of the anti inflammatory cytokines interleukin 10 (IL-10) and IL-13 can ameliorate the inflammatory response and significantly reduce hepatocellular injury. The purpose of the present study was to determine if IL-10 and IL-13 function as endogenous regulators of the hepatic inflammatory response to ischemia/reperfusion. Wild type, IL-10-, and IL-13-deficient (IL-10(-/-), IL-13(-/-)) mice were exposed to 90 minutes of partial hepatic ischemia and up to 24 hours of reperfusion. In wild type mice, expression of IL-10 and IL-13 shared similar expression profiles with maximal production after 8 hours of reperfusion. There were no significant differences between wild-type and IL-10(-/-) mice in response to hepatic ischemia and reperfusion. IL-13(-/-) mice had much greater liver injury, as assessed biochemically and histologically, than wild-type mice. There were no differences between wild-type and IL-13(-/-) mice in their production of inflammatory cytokines, but IL-13(-/-) mice displayed disrupted neutrophil accumulation, with less neutrophils present in the hepatic parenchyma and far more neutrophils adherent to the endothelium of large hepatic venules than wild-type mice. These observations were associated with increased liver endothelial cell injury in IL 13(-/-) mice, as measured by serum levels of hyaluronic acid. In vitro, IL-13 protected hepatocytes from H(2)O(2)-induced cytotoxicity. In conclusion, IL-10 is not an important endogenous regulator of the inflammatory response to hepatic ischemia/reperfusion. In contrast, endogenous IL-13 appears to be critical for the control of this response, with prominent protective effects on hepatocytes and hepatic endothelial cells. PMID- 12540781 TI - Effect of cold preservation on intracellular calcium concentration and calpain activity in rat sinusoidal endothelial cells. AB - This study was performed to determine the role of intracellular calcium concentration and calpain activity on the cellular events that occur in rat sinusoidal endothelial cells (SEC) in the cold. Intracellular calcium concentrations were measured in isolated cold preserved rat SEC. Dantrolene or 1,2-bis(o-Aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetra(acetoxymethyl) ester (BAPTA-AM) was added in some studies. In other studies, calpain activity and m-calpain and mu-calpain expression were measured during cold preservation in the presence or absence of calpain inhibitors. The effect of addition of dantrolene to preservation solutions on function of whole livers after preservation was determined. Cold preservation caused an increase in intracellular calcium concentration first detected at 1 hour of preservation. This was associated with cell rounding and actin disassembly. Dantrolene and BAPTA-AM prevented the increase in intracellular calcium concentration and reduced cell rounding and actin disassembly. Cold preservation also resulted in increased calpain activity and expression on SEC. Calpain expression was reduced by dantrolene. Calpain inhibitors N-acetyl-leu-leu-norleucinal (ALLN) and N acetyl-leu-leu-methioninal (ALLM) reduced calpain activity and expression and restored SEC cell shape and actin morphology. Dantrolene improved function of livers preserved in Eurocollins (EC) solution when tested on the isolated perfused rat liver (IPRL). In conclusion, exposure of SEC to cold results sequentially in elevated intracellular calcium concentration, increased calpain activity, and actin disassembly. PMID- 12540782 TI - The nitric oxide donor, V-PYRRO/NO, protects against acetaminophen-induced hepatotoxicity in mice. AB - The liver-selective nitric oxide (NO) donor, O(2)-vinyl 1-(pyrrolidin-1-yl)diazen 1-ium-1,2-diolate (V-PYRRO/NO), is metabolized by P-450 enzymes to release NO in the liver, and is shown to protect the liver from tumor necrosis factor alpha (TNF-alpha)-induced apoptosis and D-glactosamine/endotoxin-induced hepatotoxicity. This study was undertaken to examine the effects of V-PYRRO/NO on acetaminophen-induced hepatotoxicity in mice. Mice were given V-PYRRO/NO via osmotic pumps (1.8-5.4 mg/mL, 8 microL/h) 4 to 16 hours before a hepatotoxic dose of acetaminophen (600 mg/kg, intraperitoneally [ip]). V-PYRRO/NO administration dramatically reduced acetaminophen-induced hepatotoxicity in a dose- and time dependent manner, as evidenced by reduced serum alanine aminotransferase (ALT) activity, reduced hepatic congestion, apoptosis, and improved hepatocellular pathology. The protection afforded by V-PYRRO/NO does not appear to be caused by a decrease in the formation of toxic acetaminophen metabolites, which consumes glutathione (GSH), because V-PYRRO/NO did not alter acetaminophen-induced hepatic GSH depletion. Acetaminophen-induced lipid peroxidation, as determined by the concentrations of 4-hydroxyalkenals (4-HNE) and malondialdehyde (MDA), was reduced significantly by V-PYRRO/NO treatment. Although pretreatment was most effective, administration of V-PYRRO/NO simultaneously with acetaminophen also was able to reduce acetaminophen hepatotoxicity. Genomic analysis of the liver samples 10 hours after acetaminophen intoxication showed the enhanced expression of genes associated with stress/oxidative stress, apoptosis/cell death, and DNA damage/repair. Acetaminophen-induced alterations in gene expression were attenuated significantly by V-PYRRO/NO. Real-time reverse-transcription polymerase chain reaction (RT-PCR) and Western-blot analysis confirmed microarray results. In conclusion, V-PYRRO/NO is effective in blocking acetaminophen-induced hepatotoxicity in mice. This protection may involve the reduction of oxidative stress, the inhibition of apoptosis, and possibly the maintenance of hepatic vasculature to prevent congestion. PMID- 12540783 TI - Benzodiazepine-induced protein tyrosine nitration in rat astrocytes. AB - Recent studies indicate that ammonia and hypoosmotic astrocyte swelling can induce protein tyrosine nitration (PTN) in astrocytes with potential pathogenetic relevance for hepatic encephalopathy (HE). Because HE episodes are known to be precipitated also by sedatives, the effects of benzodiazepines on PTN in cultured rat astrocytes and rat brain in vivo were studied. In cultured rat astrocytes, diazepam, PK11195, Ro5-4864, and the benzodiazepine binding inhibitor (DBI), which acts on peripheral-type benzodiazepine receptors, induced PTN. Clonazepam, a specific ligand of the central benzodiazepine receptor, failed to induce PTN. Nanomolar concentrations of DBI and PK11195 were sufficient to increase PTN, and diazepam effects were already observed at concentrations of 1 micromol/L. Diazepam-induced PTN was insensitive to NOS inhibition and uric acid but was blunted by MK-801, BAPTA-AM, W13, and catalase, suggesting an involvement of NMDA receptor activation, elevation of the cytosolic Ca(2+) concentration [Ca(2+)](i), and hydrogen peroxide. Diazepam induced a plateau-like increase in [Ca(2+)](i) and the generation of reactive oxygen intermediates (ROIs), which are both blunted by MK-801 and BAPTA-AM. The expression of functional N-methyl-D-aspartate (NMDA) receptors on cultured rat astrocytes was confirmed by reverse transcriptase polymerase chain reaction, Western blot analysis, immunhistochemistry, and receptor autoradiography. Astroglial PTN is also found in brains from rats challenged with diazepam, indicating the in vivo relevance of the present findings. In conclusion, production of ROIs and increased PTN by benzodiazepines may alter astrocyte function and thereby contribute to the precipitation of HE episodes. PMID- 12540784 TI - Probiotics and antibodies to TNF inhibit inflammatory activity and improve nonalcoholic fatty liver disease. AB - Ob/ob mice, a model for nonalcoholic fatty liver disease (NAFLD), develop intestinal bacterial overgrowth and overexpress tumor necrosis factor alpha (TNF alpha). In animal models for alcoholic fatty liver disease (AFLD), decontaminating the intestine or inhibiting TNF-alpha improves AFLD. Because AFLD and NAFLD may have a similar pathogenesis, treatment with a probiotic (to modify the intestinal flora) or anti-TNF antibodies (to inhibit TNF-alpha activity) may improve NAFLD in ob/ob mice. To evaluate this hypothesis, 48 ob/ob mice were given either a high-fat diet alone (ob/ob controls) or the same diet + VSL#3 probiotic or anti-TNF antibodies for 4 weeks. Twelve lean littermates fed a high fat diet served as controls. Treatment with VSL#3 or anti-TNF antibodies improved liver histology, reduced hepatic total fatty acid content, and decreased serum alanine aminotransferase (ALT) levels. These benefits were associated with decreased hepatic expression of TNF-alpha messenger RNA (mRNA) in mice treated with anti-TNF antibodies but not in mice treated with VSL#3. Nevertheless, both treatments reduced activity of Jun N-terminal kinase (JNK), a TNF-regulated kinase that promotes insulin resistance, and decreased the DNA binding activity of nuclear factor kappaB (NF-kappaB), the target of IKKbeta, another TNF regulated enzyme that causes insulin resistance. Consistent with treatment related improvements in hepatic insulin resistance, fatty acid beta-oxidation and uncoupling protein (UCP)-2 expression decreased after treatment with VSL#3 or anti-TNF antibodies. In conclusion, these results support the concept that intestinal bacteria induce endogenous signals that play a pathogenic role in hepatic insulin resistance and NAFLD and suggest novel therapies for these common conditions. PMID- 12540785 TI - Uroporphyria caused by ethanol in Hfe(-/-) mice as a model for porphyria cutanea tarda. AB - Two major risk factors for the development of porphyria cutanea tarda (PCT) are alcohol consumption and homozygosity for the C282Y mutation in the hereditary hemochromatosis gene (HFE). To develop an animal model, Hfe knockout mice were treated continuously with 10% ethanol in drinking water. By 4 months, uroporphyrin (URO) was detected in the urine. At 6 to 7 months, hepatic URO was increased and hepatic uroporphyrinogen decarboxylase (UROD) activity was decreased. Untreated Hfe(-/-) mice or wild-type mice treated with or without ethanol did not show any of these biochemical changes. Treatment with ethanol increased hepatic nonheme iron and hepatic 5-aminolevulinate synthase activity in Hfe(-/-) but not wild-type mice. The increases in nonheme iron in Hfe(-/-) mice were associated with diffuse increases in iron staining of parenchymal cells but without evidence of significant liver injury. In conclusion, the results of this study suggest that the uroporphyrinogenic effect of ethanol is mediated by its effects on hepatic iron metabolism. Ethanol-treated Hfe(-/-) mice seem to be an excellent model for studies of alcohol-mediated PCT. PMID- 12540786 TI - Nadolol plus spironolactone in the prophylaxis of first variceal bleed in nonascitic cirrhotic patients: A preliminary study. AB - Treatment with beta-blockers fails to decrease portal pressure in nearly 40% of cirrhotic patients. Recent studies have suggested that treatment with spironolactone reduces pressure and flow in the portal and variceal systems. This trial was designed to assess if nadolol plus spironolactone is more effective than nadolol alone to prevent the first variceal bleeding. One hundred patients with medium and large varices who had never bled and were without ascites were included in a prospective, randomized, multicenter, double-blind, placebo controlled trial. The patients were randomized into 2 groups: 51 received nadolol plus placebo (N + P) and 49 received nadolol plus spironolactone 100 mg/d (N + S). Hepatic venous pressure gradient (HVPG) and activity of the renin-aldosterone system (plasma renin activity/plasma aldosterone levels) were measured in 24 patients. There were no significant differences in the appearance of variceal bleeding and ascites between groups at a mean follow-up of 22 +/- 16 months. However, analyzing both complications together, the incidence was significantly higher in the N + P group than in the N + S group (39% vs. 20%; P <.04). Clinical ascites was also higher in patients in the N + P group than in the N + S group (21% vs. 6%; P <.04). Significant increases in plasma renin activity and plasma aldosterone levels were only observed in patients in the N + S group (P <.01). The cumulative probabilities of remaining free of bleeding and ascites were similar in both groups after 70 months of follow-up. In conclusion, these results suggest that nadolol plus spironolactone does not increase the efficacy of nadolol alone in the prophylaxis of the first variceal bleeding. However, when bleeding and ascites were considered together, the combined therapy effectively reduced the incidence of both portal-hypertensive complications. PMID- 12540787 TI - Endoscopic screening for esophageal varices in cirrhosis: Is it ever cost effective? AB - Current guidelines for the management of patients with compensated cirrhosis recommend universal screening endoscopy followed by prophylactic beta-blocker therapy to prevent initial hemorrhage in those found to have esophageal varices. However, the cost-effectiveness of this recommendation has not been established. Our objective was to determine whether screening endoscopy is cost-effective compared with empiric medical management in patients with compensated cirrhosis. Decision analysis with Markov modeling was used to calculate the cost effectiveness of 6 competing strategies: (1) universal screening endoscopy (EGD) followed by beta-blocker (BB) therapy (EGD-->BB) if varices are present, (2) EGD followed by endoscopic band ligation (EBL) (EGD-->EBL) if varices are present, (3) selective screening endoscopy (sEGD) in high risk patients followed by BB therapy if varices are present (sEGD-->BB), (4) selective screening endoscopy followed by EBL (sEGD-->EBL) if varices are present, (5) empiric beta-blocker therapy in all patients, and (6) no prophylactic therapy ("Do Nothing"). Cost estimates were from a third-party payer perspective. The main outcome measure was the cost per initial variceal hemorrhage prevented. The "Do Nothing" strategy was the least expensive yet least effective approach. Compared with the "Do Nothing" strategy, the empiric beta-blocker strategy cost an incremental $12,408 per additional variceal bleed prevented. Compared with the empiric beta-blocker strategy, in turn, both the EGD-->BB and the EGD-->EBL strategies cost over $175,000 more per additional bleed prevented. The sEGD-->BB and sEGD-->EBL strategies were more expensive and less effective than the empiric beta-blocker strategy. In conclusion, empiric beta-blocker therapy for the primary prophylaxis of variceal hemorrhage is a cost-effective measure, as the use of screening endoscopy to guide therapy adds significant cost with only marginal increase in effectiveness. PMID- 12540788 TI - Low doses of isosorbide mononitrate attenuate the postprandial increase in portal pressure in patients with cirrhosis. AB - Postprandial hyperemia is associated with a significant increase in portal pressure in cirrhosis, which may contribute to progressive dilation and rupture of gastroesophageal varices. In cirrhosis, an insufficient hepatic production of nitric oxide (NO) may impair the expected hepatic vasodilatory response to increased blood flow, further exaggerating the postprandial increase in portal pressure. This study was aimed at investigating whether low doses of an oral NO donor might counteract the postprandial peak in portal pressure. Twenty-three portal hypertensive cirrhotics, 8 of them under propranolol therapy, were randomized to receive orally 5-isosorbide mononitrate (ISMN; 10 mg; n = 11) or placebo (n = 12) and a standard liquid meal 15 minutes later. Hepatic venous pressure gradient (HVPG), mean arterial pressure (MAP), and hepatic blood flow (HBF) were measured at baseline and 15, 30, and 45 minutes after a meal. ISMN significantly attenuated the postprandial increase in portal pressure as compared with placebo (peak HVPG increase: 2.4 +/- 1.4 mm Hg vs. 5.2 +/- 2.1 mm Hg, P =.002). Percentual increases in HBF were similar in both groups. MAP decreased slightly in ISMN group (-7.5% +/-.5%; P <.01 vs. baseline). These effects were also observed in patients on chronic propranolol therapy. In conclusion, hepatic NO supplementation by low doses of ISMN effectively reduces the postprandial increase of portal pressure in cirrhosis, with only a mild effect on arterial pressure. The same was observed in patients receiving propranolol. Our results suggest that therapeutic strategies based on selective hepatic NO delivery may improve the treatment of portal hypertension. PMID- 12540789 TI - Functional significance of hepatic arterial flow reserve in patients with cirrhosis. AB - In cirrhosis, hepatic arterial vasodilatation occurs in response to reduced portal venous blood flow. However, although the hepatic arterial flow reserve is high in patients with cirrhosis, its impact on hepatic function is unknown. This study investigated the effect of adenosine-induced hepatic arterial vasodilatation on different markers of liver function. In 20 patients with cirrhosis (Child-Pugh class A/B/C: n = 2/7/11) adenosine (2-30 microg x min(-1) x kg body wt(-1)) was infused into the hepatic artery and hepatic arterial average peak flow velocities (APV), pulsatility indices (PI), and blood flow volumes (HABF) were measured using digital angiography and intravascular Doppler sonography. Indocyanine green (ICG), lidocaine, and galactose were administered intravenously in doses of 0.5, 1.0, and 500 mg/kg body weight in the presence of adenosine-induced hepatic arterial vasodilatation and, on a separate study day, without adenosine. ICG disappearance, galactose elimination capacity (GEC), and formation of the lidocaine metabolite monoethylglycinxylidide (MEGX) were assessed. Adenosine markedly increased APV and HABF and markedly decreased PI. Serum MEGX concentrations were 63.7 +/- 18.2 (median, 62; range, 36-107) and 99.0 +/- 46.3 (82.5; 49-198) ng/mL in the absence and presence of adenosine infusion, respectively (P =.001). Adenosine-induced changes in MEGX concentrations were correlated inversely to changes in APV (r = -0.5, P =.02) and PI (r = -0.55, P =.01) and were more marked in Child-Pugh class C compared with Child-Pugh class A patients (57.4 +/- 49.9 [44; -14 to 140] vs. 8.4 +/- 16.5 [13; -11 to 35] ng/mL, P <.01). In conclusion, hepatic arterial vasodilatation provides substantial functional benefit in patients with cirrhosis. The effect does not depend directly on hepatic arterial macroperfusion and is observed preferentially in patients with decompensated disease. PMID- 12540790 TI - Cardiac hepatopathy: clinical, hemodynamic, and histologic characteristics and correlations. AB - Cardiac hepatopathy, hepatic injury caused by cardiac dysfunction, is a common entity but has been characterized incompletely, particularly the relationship between hemodynamics and histology. We aimed to describe the clinical, biochemical, hemodynamic, and histologic characteristics of this disorder. Eighty three patients from 2 tertiary referral centers were studied. Patients were divided into 3 groups based on the duration of cardiac dysfunction: (1) acute (n = 12); (2) chronic (n = 53); and (3) acute on chronic (n = 18). Results showed that serum aminotransferase levels were increased typically, particularly in the acute group (median aspartate aminotransferase level was 30.2 times the upper limit of normal [range, 1-100]; P <.0001 vs. the chronic group). The most salient hemodynamic features were elevated right atrial (14 mm Hg [range, 1-29]), and hepatic venous pressures (wedged: 18 mm Hg [range, 5-35]; free: 15 mm Hg [range, 2-30]). The hepatic venous pressure gradient was normal in most (81%), correlated moderately with the aminotransferase levels (aspartate aminotransferase level: r =.59; P <.0001), and associated with the presence of centrilobular necrosis and inflammation, periportal necrosis, and stainable hepatic iron (P <.05 for all comparisons), but not fibrosis. Sinusoidal dilatation was associated with higher right atrial (P =.047) and free hepatic venous pressures (P =.06). Although cirrhosis was rare (n = 1), centrilobular fibrosis was common (74%) and not associated with any hemodynamic measurement. In conclusion, cardiac hepatopathy has diverse clinical, hemodynamic, and histologic manifestations that vary with the temporal course of cardiac dysfunction. Hepatic fibrosis is common, but does not correlate with systemic or hepatic hemodynamics. PMID- 12540791 TI - Diagnosis of portopulmonary hypertension in candidates for liver transplantation: a prospective study. AB - Portopulmonary hypertension represents a major risk factor for transplantation; therefore, preoperative detection is crucial. The aims of this study were to determine (1) whether Doppler echocardiography performed at evaluation is a reliable tool for detecting portopulmonary hypertension and (2) the incidence of acquired portopulmonary hypertension profile after evaluation. One hundred sixty five patients had Doppler echocardiography and right heart catheterization at evaluation over a 9-year period. All patients had a prospective follow-up, and the results of catheterization at evaluation were compared with those obtained at the time of transplantation. Seventeen of 165 patients met the criteria for portopulmonary hypertension on Doppler echocardiography. Portopulmonary hypertension was confirmed by catheterization in 10 patients and ruled out in 7. There were no false negatives for echocardiography. Mean pulmonary artery pressure was significantly higher during the initial phase of transplantation than at evaluation (17.8 +/- 4.3 vs. 20.3 +/- 5.5 mm Hg, respectively, P <.0001), and there was no significant correlation between values obtained at these 2 time points. Three patients showed to have acquired portopulmonary hypertension profile while waiting for a graft within time intervals ranging from 2.5 to 5 months. In conclusion, Doppler echocardiography is a highly sensitive tool for detecting portopulmonary hypertension. However, because this technique has a poor positive predictive value, right heart catheterization is recommended for confirming portopulmonary hypertension. In addition, the absence of portopulmonary hypertension at evaluation does not exclude the occasional occurrence of acquired portopulmonary hypertension profile after listing. PMID- 12540792 TI - Genetic and epigenetic factors in autoimmune reactions toward cytochrome P4502E1 in alcoholic liver disease. AB - Autoimmune reactions are often associated with alcoholic liver disease; however, the mechanisms responsible are largely unknown. This study investigates the potential role of the immune response against hydroxyethyl free radical (HER) derived antigens and of polymorphisms in immunoregulatory genes in the development of anti-cytochrome P4502E1 (CYP2E1) autoantibodies in alcohol abusers. Immunoglobulin G (IgG) recognizing human CYP2E1 and HER-derived epitopes were measured by microplate immunosorbent assay in the sera of 90 patients with alcoholic fibrosis/cirrhosis (ALD), 37 heavy drinkers without liver disease or steatosis only (HD), and 59 healthy subjects. Single nucleotide polymorphisms in the interleukin 10 (IL-10) promoter and in exon 1 of the cytotoxic T-lymphocyte antigen-4 (CTLA-4) gene were genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis. The titers and frequency of anti-CYP2E1 autoantibodies were significantly higher in ALD than in HD subjects or controls. ALD patients with anti-HER IgG had higher titers and a 4-fold increased risk (OR: 4.4 [1.8-10.9]) of developing anti-CYP2E1 autoantibodies than subjects without anti-HER antibodies. The mutant CTLA-4 G allele, but not the IL-10 polymorphism, was associated with an enhanced risk of developing anti-CYP2E1 IgG (OR: 3.8 [1.4 10.3]). CTLA-4 polymorphism did not influence antibody formation toward HER antigens. ALD patients with concomitant anti-HER IgG and the CTLA-4 G allele had a 22-fold higher (OR: 22.9 [4.2-125.6]) risk of developing anti-CYP2E1 autoreactivity than subjects negative for these factors. In conclusion, antigenic stimulation by HER-modified CYP2E1 combined with an impaired control of T-cell proliferation by CTLA-4 mutation promotes the development of anti-CYP2E1 autoantibodies that might contribute to alcohol-induced liver injury. PMID- 12540793 TI - Selective increase of brain lactate synthesis in experimental acute liver failure: results of a [H-C] nuclear magnetic resonance study. AB - Acute liver failure (ALF) results in alterations of energy metabolites and of glucose-derived amino acid neurotransmitters in brain. However, the dynamics of changes in glucose metabolism remain unclear. The present study was undertaken using (1)H and (13)C nuclear magnetic resonance (NMR) spectroscopy to determine the rates of incorporation of glucose into amino acids and lactate via cell specific pathways in relation to the severity of encephalopathy and brain edema in rats with ALF because of hepatic devascularization. Early (precoma) stages of encephalopathy were accompanied by significant 2- to 4.5-fold (P <.001) increases of total brain glutamine and lactate concentrations. More severe (coma) stages of encephalopathy and brain edema led to a further significant increase in brain lactate but no such increase in glutamine. Furthermore, (13)C isotopomer analysis showed a selective increase of de novo synthesis of lactate from [1-(13)C]glucose resulting in 2.5-fold increased fractional (13)C enrichments in lactate at coma stages. [2-(13)C]glutamine, synthesized through the astrocytic enzyme pyruvate carboxylase, increased 10-fold at precoma stages but showed no further increase at coma stages of encephalopathy. (13)C-label incorporation into [4 (13)C]glutamate, synthesized mainly through neuronal pyruvate dehydrogenase, was selectively reduced at coma stages, whereas brain GABA synthesis was unchanged at all time points. In conclusion, increased brain lactate synthesis and impaired glucose oxidative pathways rather than intracellular glutamine accumulation are the major cause of brain edema in ALF. Future NMR spectroscopic studies using stable isotopes and real-time measurements of metabolic rates could be valuable in the elucidation of the cerebral metabolic consequences of ALF in humans. PMID- 12540794 TI - Systematic review of randomized trials for unresectable hepatocellular carcinoma: Chemoembolization improves survival. AB - There is no standard treatment for patients with unresectable hepatocellular carcinoma (HCC). Survival benefits derived from medical interventions are controversial. The aim of this systematic review was to assess the evidence of the impact of medical treatments on survival. Randomized controlled trials (RCTs) that were published as full papers assessing survival for primary treatments of HCC were included. MEDLINE, the Cochrane Library, CANCERLIT, and a manual search from 1978 to May 2002 were used. The primary end point was survival, and the secondary end point was response to treatment. Estimates of effect were calculated according to the random effects model. Sensitivity analysis included methodological quality. We identified 61 randomized trials, but only 14 met the criteria to perform a meta-analysis assessing arterial embolization (7 trials, 545 patients) or tamoxifen (7 trials, 898 patients). Arterial embolization improved 2-year survival compared with control (odds ratio [OR], 0.53; 95% confidence interval [CI], 0.32-0.89; P =.017). Sensitivity analysis showed a significant benefit of chemoembolization with cisplatin or doxorubicin (OR, 0.42; 95% CI, 0.20-0.88) but none with embolization alone (OR, 0.59; 95% CI, 0.29 1.20). Overall, treatment induced objective responses in 35% of patients (range, 16%-61%). Tamoxifen showed no antitumoral effect and no survival benefits (OR, 0.64; 95% CI, 0.36-1.13; P =.13), and only low-quality scale trials suggested 1 year improvement in survival. In conclusion, chemoembolization improves survival of patients with unresectable HCC and may become the standard treatment. Treatment with tamoxifen does not modify the survival of patients with advanced disease. PMID- 12540795 TI - Adherence and mental side effects during hepatitis C treatment with interferon alfa and ribavirin in psychiatric risk groups. AB - Psychiatric disorders or drug addiction are often regarded as contraindications against the use of interferon alfa (IFN-alpha) in patients with chronic hepatitis C. Our aim was to obtain prospective data on adherence to as well as efficacy and mental side effects of treatment with IFN-alpha in different psychiatric risk groups compared with controls. In a prospective trial, 81 patients with chronic hepatitis C (positive hepatitis C virus[HCV] RNA and elevated alanine aminotransferase[ALT] level) and psychiatric disorders (n = 16), methadone substitution (n = 21), former drug addiction (n = 21), or controls without a psychiatric history or drug addiction (n = 23) were treated with a combination of IFN-alpha-2a 3 MU 3 times weekly and ribavirin (1,000-1,200 mg/d). Sustained virologic response (overall, 37%) did not differ significantly between subgroups. No significant differences between groups were detected with respect to IFN-alpha related development of depressions during treatment. However, in the psychiatric group, significantly more patients received antidepressants before and during treatment with IFN-alpha (P <.001). Most of those who dropped out of the study were patients with former drug addiction (43%; P =.04) compared with 14% in the methadone group, 13% in the control group, and 18% in the psychiatric group. No patient in the psychiatric group had to discontinue treatment because of psychiatric deterioration. In conclusion, our data do not confirm the supposed increased risk for IFN-alpha-induced mental side effects and dropouts in psychiatric patients if interdisciplinary care and antidepressant treatment are available. Preexisting psychiatric disorders or present methadone substitution should no longer be regarded as contraindications to treatment of chronic hepatitis C with IFN-alpha and ribavirin in an interdisciplinary setting. PMID- 12540796 TI - Modulation of cellular immune response against hepatitis C virus nonstructural protein 3 by cationic liposome encapsulated DNA immunization. AB - A vaccine strategy directed to increase Th1 cellular immune responses, particularly to hepatitis C virus (HCV) nonstructural protein 3 (NS3), has considerable potential to overcome the infection with HCV. DNA vaccination can induce both humoral and cellular immune responses, but it became apparent that the cellular uptake of naked DNA injected into muscle was not very efficient, as much of the DNA is degraded by interstitial nucleases before it reaches the nucleus for transcription. In this paper, cationic liposomes composed of different cationic lipids, such as dimethyl-dioctadecylammonium bromide (DDAB), 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), or 1,2-dioleoyl-sn-glycerol-3 ethylphosphocholine (DOEPC), were used to improve DNA immunization in mice, and their efficiencies were compared. It was found that cationic liposome-mediated DNA immunization induced stronger HCV NS3-specific immune responses than immunization with naked DNA alone. Cationic liposomes composed of DDAB and equimolar of a neutral lipid, egg yolk phosphatidylcholine (EPC), induced the strongest antigen-specific Th1 type immune responses among the cationic liposome investigated, whereas the liposomes composed of 2 cationic lipids, DDAB and DOEPC, induced an antigen-specific Th2 type immune response. All cationic liposomes used in this study triggered high-level, nonspecific IL-12 production in mice, a feature important for the development of maximum Th1 immune responses. In conclusion, the cationic liposome-mediated gene delivery is a viable HCV vaccine strategy that should be further tested in the chimpanzee model. PMID- 12540797 TI - A recombinant adenovirus encoding hepatitis C virus core and E1 proteins protects mice against cytokine-induced liver damage. AB - Hepatitis C virus (HCV) infection has a strong tendency to evolve to chronicity despite up-regulation of proapoptotic cytokines in the inflamed liver. The mechanisms responsible for persistent viral replication in this inflammatory environment are obscure. It is conceivable that viral replication would be facilitated if the infected hepatocytes are rendered resistant to cytokine induced cytotoxicity. In this study, we investigated if an adenovirus encoding HCV core and E1 (RAdCE1) could reduce liver cell injury in different in vivo models of cytokine-mediated hepatotoxicity in mice. We show that RAdCE1 markedly attenuates hepatocellular apoptosis and the increase in serum transaminase levels after concanavalin A (con A) challenge. This protective effect is accompanied by an inhibition of nuclear translocation of nuclear factor kappaB (NF-kappaB); reduced expression of inducible nitric oxide synthase (iNOS); decreased hepatic messenger RNA levels of chemokines macrophage inflammatory protein 2 (MIP-2), monocyte chemoattractant protein 1 (MCP-1), and interferon-inducible protein 10 (IP-10); and abrogation of liver leukocyte infiltration. RAdCE1 also causes a reduction in serum transaminase levels and inhibits hepatocellular apoptosis in mice given tumor necrosis factor (TNF)-alpha plus D-galactosamine. In conclusion, HCV structural antigens can protect liver cells against the proapoptotic effects of proinflammatory cytokines. The antiapoptotic status of infected liver cells may represent a mechanism favoring viral persistence. Our findings also suggest that, in chronic hepatitis C, the burden of hepatocellular damage mainly affects noninfected liver cells. PMID- 12540798 TI - A new approach to therapy for hepatocellular carcinoma? PMID- 12540799 TI - Therapy for primary prophylaxis of varices: and, the winner is...? PMID- 12540801 TI - Octreotide treatment of hepatocellular carcinoma. PMID- 12540804 TI - Applicability of real-time PCR combined with melting curve analysis for the study of hepatitis B virus genome mutations. PMID- 12540806 TI - Genetic alterations in hepatocellular adenomas. PMID- 12540808 TI - Higher frequency of HCV in patients with non-Hodgkin lymphoma: is it enough to suggest an association with B-cell NHL? PMID- 12540810 TI - Fractal analysis in liver fibrosis. PMID- 12540812 TI - Chopin's serpin. PMID- 12540813 TI - Surface and buildup dose characteristics for 6, 10, and 18 MV photons from an Elekta Precise linear accelerator. AB - Understanding head scatter characteristics of photon beams is vital to properly commission treatment planning (TP) algorithms. Simultaneously, having definitive surface and buildup region dosimetry is important to optimize bolus. The Elekta Precise linacs have unique beam flattening filter configurations for each photon beam (6, 10, and 18 MV) in terms of material and location. We performed a comprehensive set of surface and buildup dose measurements with a thin window parallel-plate (PP) chamber to examine effects of field size (FS), source-to-skin distance (SSD), and attenuating media. Relative ionization data were converted to fractional depth dose (FDD) after correcting for bias effects and using the Gerbi method to account for chamber characteristics. Data were compared with a similar vintage Varian linac. At short SSDs the surface and buildup dose characteristics were similar to published data for Varian and Elekta accelerators. The FDD at surface (FDD(0)) for 6, 10, and 18 MV photons was 0.171, 0.159, and 0.199, respectively, for a 15x15 cm2, 100 cm SSD field. A blocking tray increased FDD(0) to 0.200, 0.200, and 0.256, while the universal wedge decreased FDD(0) to 0.107, 0.124, and 0.176. FDD(0) increased linearly with FS (approximately 1.16%/cm). FDD(0) decreased exponentially for 10 and 18 MV with increasing SSD. However, the 6 MV FDD(0) actually increased slightly with increasing SSD. This is likely due to the unique distal flattening filter for 6 MV. The measured buildup curves have been used to optimize TP calculations and guide bolus decisions. Overall the FDD(0) and buildup doses were very similar to published data. Of interest were the relatively low 10 MV surface doses, and the 6 MV FDD(0)'s dependence on SSD. PMID- 12540814 TI - Multimodality image fusion in dose escalation studies of brain tumors. AB - This article examines the utility of integrating images from computed tomography (CT), magnetic resonance imaging (MRI), and single photon emission computed tomography (SPECT) for radiation treatment planning of brain tumors for dose escalation studies. The information obtained from these imaging modalities is complementary to each other and could provide anatomic (through CT and MRI) and metabolic (through SPECT) information of the target. This anato-metabolic target localization could be expected to facilitate precise radiation therapy planning for brain tumors by delineating the boundary between the tumor, edema, and the normal brain parenchyma and identify the viable tumor nidus with greater degree of certainty. This could in turn lead to minimize dose to the normal tissue and permit dose escalation to the region of interest. The utility of these anato metabolic imaging modalities for defining the clinical target volumes along with planning target volumes for different phases of the radiation therapy is illustrated. PMID- 12540815 TI - Assessment of consistency in contouring of normal-tissue anatomic structures. AB - The purpose of this work is to estimate the uncertainty in the manual contouring of normal anatomical structures. The heart, esophagus, and spinal cord were contoured manually on six sets of computed tomography images by six dosimetrists whose experience ranged from 1 year to over 15 years. To determine the differences between inter- and intraobserver variations, each data set was contoured by one of the dosimetrists five times and once each by the five other dosimetrists. The magnitude of the discrepancies in delineating the contours was assessed. Intradosimetrist contouring discrepancies were as follows: esophagus, average 0.3 cm and maximum 2.9 cm; heart, average 0.5 cm and maximum 7.6 cm; and spinal cord, average 0.1 cm and maximum 0.7 cm. Interdosimetrist contouring discrepancies were as follows: esophagus, average 0.4 cm and maximum 3.1 cm; heart, average 0.7 cm and maximum 8.1 cm; and spinal cord, average 0.2 cm and maximum 0.9 cm. Significant discrepancies can occur when normal anatomic structures are contoured manually. Interdosimetrist discrepancies are typically slightly greater than intradosimetrist discrepancies. The magnitude of the discrepancies does not appear to be correlated to the experience of the dosimetrist. PMID- 12540816 TI - Dosimetric comparison of extended dose range film with ionization measurements in water and lung equivalent heterogeneous media exposed to megavoltage photons. AB - In this study, a dosimetric evaluation of the new Kodak extended dose range (EDR) film versus ionization measurements has been conducted in homogeneous solid water and water-lung equivalent layered heterogeneous phantoms for a relevant range of field sizes (up to a field size of 25x25 cm2 and a depth of 15 cm) for 6 and 15 MV photon beams from a linear accelerator. The optical density of EDR film was found to be linear up to about 350 cGy and over-responded for larger fields and depths (5% for 25x25 cm2 at depth of 15 cm compared to a 10x10 cm2, 5 cm depth reference value). Central axis depth dose measurements in solid water with the film in a perpendicular orientation were within 2% of the Wellhofer IC-10 measurements for the smaller field sizes. A maximum discrepancy of 8.4% and 3.9% was found for the 25x25 cm2 field at 15 cm depth for 6 and 15 MV photons, respectively (with curve normalization at a depth of 5 cm). Compared to IC-10 measurements, film measured central axis depth dose inside the lung slab showed a slight over-response (at most 2%). At a depth of 15 cm in the lung phantom the over-response was found to be 7.4% and 3.7% for the 25x25 cm2 field for 6 and 15 MV photons, respectively. When results were presented as correction factors, the discrepancy between the IC-10 and the EDR was greatest for the lowest energy and the largest field size. The effect of the finite size of the ion chamber was most evident at smaller field sizes where profile differences versus film were observed in the penumbral region. These differences were reduced at larger field sizes and in situations where lateral electron transport resulted in a lateral spread of the beam, such as inside lung material. Film profiles across a lung tumor geometry phantom agreed with the IC-10 chamber within the experimental uncertainties. From this investigation EDR film appears to be a useful medium for relative dosimetry in higher dose ranges in both water and lung equivalent material for moderate field sizes and depths. PMID- 12540817 TI - Patient specific quality assurance for the delivery of intensity modulated radiotherapy. AB - A patient specific quality assurance program has been developed to facilitate the clinical implementation of intensity modulated radiotherapy (IMRT) delivered using a micro-multileaf collimator. The methodology includes several dosimetric tasks that are performed prior to the treatment of each patient. Film dosimetry is performed for each individual field and for the multifield composite plan. Individual field measurements are performed at a depth of 5 cm in a water equivalent slab phantom; export of dose calculations from the treatment planning system is similarly specified. For the composite distribution, parameters from the patient plan are applied to an IMRT phantom, and film is exposed in an axial orientation. Distributions are compared with the aid of software developed for the specific tasks. The measured and calculated dose distributions can be superimposed and positioned graphically using move, rotate, and mirror tools, as well as by specifying isocenter coordinates and using fiducial marks. Horizontal and vertical profiles are available for analysis. Dose difference, distance-to agreement, and gamma index, the minimum scaled multidimensional distance between a measurement and a calculation point determined in combined dose and physical distance space, are calculated along a specified isodose line and displayed. gamma provides an excellent measure of disagreement between measurement and calculation for complex intensity distributions. We specify 3% dose difference and 3 mm distance as our scaling acceptability criteria. Absolute dosimetry for each composite plan is performed using an ionization chamber. To date, excellent agreement between measurements and calculations has been observed. PMID- 12540818 TI - Corrections to traditional methods of verifying tangential-breast 3D monitor-unit calculations: use of an equivalent triangle to estimate effective fields. AB - This paper describes an innovative method for correctly estimating the effective field size of tangential-breast fields. The method uses an "equivalent triangle" to verify intact breast tangential field monitor-unit settings calculated by a 3D planning system to within 2%. The effects on verification calculations of loss of full scatter due to beam oblique incidence, proximity to field boundaries, and reduced scattering volumes are handled properly. The methodology is validated by comparing calculations performed by the 3D planning system with the respective verification estimates. The accuracy of this technique is established for dose calculations both with and without heterogeneity corrections. PMID- 12540819 TI - Wire or coated balloon? Searching for an optimal source for intravascular brachytherapy with beta emitters using (32)P as an example. AB - This study identifies basic dosimetric differences between two designs for intravascular brachytherapy (IVBT) in current clinical practice and ongoing trials and their clinical implications within beta emitting systems using P-32 as an example. The two designs are (i) the wire-type source, where the radioactive source material is confined to a wirelike structure within the vessel lumen, and (ii) the balloon-surface source, where the radioactive source material is distributed over a surface area (balloon-wall) which is brought in close proximity with the vessel wall. Using Monte Carlo simulations with the EGS4 code, the target coverage, the influence of centering errors, and the perturbation of the dose distribution caused by metallic stents have been compared. The radial dose fall-off in the target region was found to be steeper for balloon surface systems compared with wire systems. The inner lumen wall dose for a balloon surface source was 25% higher than that for a wirelike source (2.5 mm vessel diameter). However, the comparably shallower fall-off from wire-type systems is very sensitive to centering uncertainties. A 0.5 mm displacement, for example, will cause the dose to change by a factor of 2 at the inner vessel wall and by a factor of 1.8 at the prescription point. It is shown that the interference from metallic stents is more significant for wire-type systems than it is for balloon surface-type systems, where double the dose variation beyond the stent at the radial prescription distance may occur. Centering uncertainties dominate the dose perturbation effects for wire-type systems. Balloon-surface-type designs show a more predictable dose distribution that features, however, a higher inner vessel surface dose. Since a direct clinical comparison of systems of both types is not likely, these findings should be considered when interpreting clinical results from treatments with either type of source and, possibly, for future source design. PMID- 12540820 TI - A seed specific dose kernel method for low-energy brachytherapy dosimetry. AB - We describe a method for independently verifying the dose distributions from pre- and post-implant brachytherapy source distributions. Monte Carlo calculations have been performed to characterize the three-dimensional dose distribution in water phantom from a low-energy brachytherapy source. The calculations are performed in a voxelized, Cartesian coordinate geometry and normalized based upon a separate Monte Carlo calculation for the seed specific air-kerma strength to produce an absolute dose grid with units of cGy hr(-1) x U(-1). The seed specific, three-dimensional dose grid is stored as a text file for processing using a separate visual basic program. This program requires the coordinate positions of each seed in the pre- or post-plan and sums the kernel file for a three-dimensional composite dose distribution. A kernel matrix size of 81x81x81 with a voxel size of 1.0x1.0x1.0 mm3 was chosen as a compromise between calculation time, kernel size, and truncation of the stored dose distribution as a function of radial distance from the midpoint of the seed. Good agreement is achieved for a representative pre- and post-plan comparison versus a commercial implementation of the TG-43 brachytherapy dosimetry protocol. PMID- 12540821 TI - Enhanced dynamic wedge factors at off-axis points in asymmetric fields. AB - Several recent reports have described methods for calculating enhanced dynamic wedge factors (EDWFs). Many of these reports use the monitor-unit (MU) fraction method to predict EDWFs as a function of field size. Although simple in approach, MU fraction methods do not produce accurate EDWFs in large or asymmetric fields. A recently described technique, based on the MU fraction method works well for large and asymmetric fields, but only when the calculation point is in the center of the field. Other existing methods based on beam-segment superposition do not have this limitation. These beam summation methods, however, are difficult to implement in routine clinical MU calculation schemes. In this paper, we present a simple calculation method that estimates EDWFs at off-axis calculation points in both symmetric and asymmetric fields. Our method, which also is based on the MU fraction method, similarly uses empirically determined field-size corrections but also applies wedged-field profiles to estimate EDWFs that are independent of calculation-point location and field symmetry. EDWF measurements for a variety of field sizes and calculation-point locations for both 6- and 18-MV x-ray beams were performed to validate our calculations and those of our ADAC Pinnacle3 Treatment Planning System. The disagreement between the calculated and measured EDWFs over the useful clinical range of field sizes and calculation-point locations was less than 2%. The worst disagreement was 3% and occurred at a point 8.5 cm from the center of an asymmetric 25 (wedged direction)x20 cm2 60 degrees wedged field. Detailed comparisons of measurements with calculations and wedge factors obtained from the ADAC Pinnacle3 Treatment Planning System will be presented. In addition, the strengths and weaknesses of this calculation method will be discussed. PMID- 12540822 TI - Interpretation of normal anatomic structures on chest radiography: comparison of Fuji computed radiography (FCR) 5501D with FCR 5000 and screen-film system. AB - The purpose of this study was to investigate the usefulness of Fuji Computed Radiography (FCR) 5501D by comparing it with FCR 5000 and a screen-film system (S/F). Posteroanterior chest radiographs of ten patients with no abnormality on chest CT scans were obtained with FCR 5501D, FCR 5000, and S/F. Six observers (three radiologists and three radio-technologists) evaluated the visibility of nine normal anatomic structures (including lungs, soft tissue, and bones) and overall visibility on each image. Observers scored using a five-point scale on each structure. FCR 5000 showed a significantly higher score in soft tissue and bone structures, and overall visibility compared with S/F, but, there was no significant difference between them in the visibility of all four normal lung structures. Compared with S/F, the score for FCR 5501D was higher in eight of the nine normal structures, including three of the four lung structures (unobscured lung, retrocardiac lung, and subdiaphragmatic lung), and overall visibility. Compared with FCR 5000, the score for FCR 5501D was higher in three normal structures, including two of the four lung structures (unobscured lung and subdiaphragmatic lung), and overall visibility. FCR 5501D was the best among the three techniques to visualize normal anatomic structures, particularly the obscured and unobscured lung. PMID- 12540823 TI - Comparison of computed radiography and film/screen combination using a contrast detail phantom. AB - The purpose of this work is to compare computed radiography (Kodak CR 400) and film/screen combination (Speed 400) systems in regards of patient dose, technique settings, and contrast-detail detectability. A special contrast-detail phantom with drilled holes of varying diameter (detail) and varying depth (contrast) was utilized. Various thicknesses of the Lucite sheets were utilized to simulate scattering tissues. Images of the phantom were acquired using a range of 60-120 kVp for film/screen and CR with a conventional x-ray tube and then for CR with additional 2 mm aluminum added filtration to the x-ray beam. The patient entrance skin dose was measured while maintaining 1.6 o.d. for film/screen images and 1900 Exposure Index for CR images. CR phantom images were displayed on the diagnostic workstation for soft copy reading as well as printed on films for hard copy reading on viewbox. Four physicists evaluated the images by scoring the threshold target depth along the row of the same target diameter. Detection ratio was calculated by counting the number of detectable targets divided by the total number of targets in the phantom. The overall score was related to the patient entrance skin dose, kVp, and the thickness of the scattering material. The patient entrance skin dose was reduced as the additional aluminum filter was added to the x-ray beam. Our findings suggested using a higher kVp setting and additional added filtration would reduce the patient entrance skin dose without compromising the contrast-detail detectability, which was compensated by the contrast manipulation on soft-copy display workstations. PMID- 12540824 TI - Blue light activates calcium-permeable channels in Arabidopsis mesophyll cells via the phototropin signaling pathway. AB - Light is a central regulator of plant growth and development. Among the processes triggered by blue and UV-A light, phototropism, stomatal movement, and chloroplast orientation rely on the activation of blue-light receptors known as phototropins. So far, these photoreceptors constitute a class of light receptor kinases unique to the plant kingdom. In Arabidopsis thaliana, the two members phot1 and phot2 have been shown to display partially overlapping functions. Up to now little is known about the signaling cascade, which links these phototropins to the physiological responses downstream of blue-light perception. Here, we show that on illumination with blue light, but not red light, voltage-dependent and calcium-permeable channels activate in the plasma membrane of mesophyll cells. Blue-light stimulation in the presence of the photosynthetic electron transport inhibitor, 3-(3,4-dichlorophenyl)-1,1-dimethylurea, indicates that blue-light receptors rather than photosynthesis control channel activity. Sensitivity toward the protein kinase inhibitor K252a further pointed to the possible involvement of light receptor kinases. In support of this hypothesis, in the photoreceptor mutant phot1-5, blue-light induction of calcium currents was dramatically reduced and was eliminated in the double mutant phot1-5 phot2-1. By contrast, in cry1-304 cry2-1, an Arabidopsis mutant lacking another class of plant blue-light receptors, the channel remained sensitive to blue light. We thus conclude that blue light triggers calcium fluxes via the phototropin-activated calcium permeable channel. PMID- 12540826 TI - Least effort and the origins of scaling in human language. AB - The emergence of a complex language is one of the fundamental events of human evolution, and several remarkable features suggest the presence of fundamental principles of organization. These principles seem to be common to all languages. The best known is the so-called Zipf's law, which states that the frequency of a word decays as a (universal) power law of its rank. The possible origins of this law have been controversial, and its meaningfulness is still an open question. In this article, the early hypothesis of Zipf of a principle of least effort for explaining the law is shown to be sound. Simultaneous minimization in the effort of both hearer and speaker is formalized with a simple optimization process operating on a binary matrix of signal-object associations. Zipf's law is found in the transition between referentially useless systems and indexical reference systems. Our finding strongly suggests that Zipf's law is a hallmark of symbolic reference and not a meaningless feature. The implications for the evolution of language are discussed. We explain how language evolution can take advantage of a communicative phase transition. PMID- 12540825 TI - Bcl-xL overexpression blocks bax-mediated mitochondrial contact site formation and apoptosis in rod photoreceptors of lead-exposed mice. AB - Photoreceptor apoptosis and resultant visual deficits occur in humans and animals with inherited and disease-, injury-, and chemical-induced retinal degeneration. A clinically relevant mouse model of progressive rod photoreceptor-selective apoptosis was produced by low-level developmental lead exposure and studied in combination with transgenic mice overexpressing Bcl-x(L) only in the photoreceptors. A multiparametric analysis of rod apoptosis and mitochondrial structure-function was performed. Mitochondrial cristae topography and connectivity, matrix volume, and contact sites were examined by using 3D electron tomography. Lead-induced rod-selective apoptosis was accompanied by rod Ca(2+) overload, rhodopsin loss, translocation of Bax from the cytosol to the mitochondria, decreased rod mitochondrial respiration and membrane potential, mitochondrial cytochrome c release, caspase-3 activation, and an increase in the number of mitochondrial contact sites. These effects occurred without mitochondrial matrix swelling, outer membrane rupture, caspase-8 activation, or Bid cleavage. Bcl-x(L) overexpression completely blocked all apoptotic events, except Ca(2+) overload, and maintained normal rod mitochondrial function throughout adulthood. This study presents images of mitochondrial contact sites in an in vivo apoptosis model and shows that Bcl-x(L) overexpression blocks increased contact sites and apoptosis. These findings extend our in vitro retinal studies with Pb(2+) and Ca(2+) and suggest that developmental lead exposure produced rod-selective apoptosis without mitochondrial swelling by translocating cytosolic Bax to the mitochondria, which likely sensitized the Pb(2+) and Ca(2+) overloaded rod mitochondria to release cytochrome c. These results have relevance for therapies in a wide variety of progressive retinal and neuronal degenerations where Ca(2+) overload, lead exposure, andor mitochondrial dysfunction occur. PMID- 12540827 TI - Innate and acquired immunity intersect in a global view of the acute-phase response. AB - Recognition of stereotypic chemical patterns by sentinel cells of the innate immune system provokes a transient deviation from homeostasis, the acute-phase response (APR). Although APR effectors have been identified individually, the complexity of the response suggested that emergent properties would be uncovered by a more comprehensive examination. Our global assessment revealed that approximately 7% of genes in the mouse are mobilized in the hepatic APR to endotoxin. Extensive metabolic adjustments include suppression of pathways for cholesterol, fatty acid, and phospholipid synthesis. Increased expression of genes for innate defense was accompanied by coordinate induction of the MHC class I antigen presentation machinery, illustrating an intersection between innate and adaptive immunity. PMID- 12540828 TI - PPARdelta is a very low-density lipoprotein sensor in macrophages. AB - Although triglyceride-rich particles, such as very low-density lipoprotein (VLDL), contribute significantly to human atherogenesis, the molecular basis for lipoprotein-driven pathogenicity is poorly understood. We demonstrate that in macrophages, VLDL functions as a transcriptional regulator via the activation of the nuclear receptor peroxisome proliferator-activated receptor delta. The signaling components of native VLDL are its triglycerides, whose activity is enhanced by lipoprotein lipase. Generation of peroxisome proliferator-activated receptor delta null macrophages verifies the absolute requirement of this transcription factor in mediating the VLDL response. Thus, our data reveal a pathway through which dietary triglycerides and VLDL can directly regulate gene expression in atherosclerotic lesions. PMID- 12540829 TI - Genetic biases for showy males: are some genetic systems especially conducive to sexual selection? AB - Male secondary sexual characters (conspicuous ornaments, signals, colors) are among nature's most striking features. Yet, it is unclear why certain groups of organisms are more likely than others to evolve these traits. One explanation for such taxonomic biases is that some genetic systems may be especially conducive to sexual selection. Here, we present theory and simulation results demonstrating that rare alleles encoding either male ornaments or female preferences for those ornaments are better protected against random loss in species with ZZZW or ZZZO sex chromosome systems (male homogamety) than in species with XXXY or XXXO systems (male heterogamety). Moreover, this protection is much stronger in diploid than haplodiploid species. We also present empirical data showing that male secondary sexual characters are better developed in diploid than haplodiploid species and in diploid species with male homogamety than in those with male heterogamety. Thus, taxonomic biases for showy males may stem from differences in sex chromosome systems. PMID- 12540830 TI - Evolution of feedback-inhibited beta /alpha barrel isoenzymes by gene duplication and a single mutation. AB - The betaalpha barrel is the common protein fold of numerous enzymes and was proposed recently to be the result of gene duplication and fusion of an ancient half-barrel. The initial enzyme of shikimate biosynthesis possesses the additional feature of feedback regulation. The crystal structure and kinetic studies on chimera and mutant proteins of yeast 3-deoxy-d-arabino-heptulosonate-7 phosphate (DAHP) synthase from Saccharomyces cerevisiae inhibited by phenylalanine (Aro3p) and DAHP synthase S. cerevisiae inhibited by tyrosine (Aro4p) give insight into important regions for regulation in the enzyme: The loop, which is connecting the two half-barrels, and structural elements added to the barrel are prerequisites for regulation and form a cavity on the N-terminal side of the betaalpha barrel. In the cavity of Aro4p at position 226 is a glycine residue, which is highly conserved in all other tyrosine-regulated DAHP synthases as well. Sequence alignments with phenylalanine-regulated DAHP synthases including Aro3p show a highly conserved serine residue at this position. An exchange of glycine to serine and vice versa leads to a complete change in the regulation pattern. Therefore the evolution of these differently feedback inhibited isoenzymes required gene duplication and a single mutation within the internal extra element. Numerous additional amino acid substitutions present in the contemporary isoenzymes are irrelevant for regulation and occurred independently. PMID- 12540831 TI - Quantitation of changes in protein phosphorylation: a simple method based on stable isotope labeling and mass spectrometry. AB - Reversible protein phosphorylation plays an important role in many cellular processes. However, a simple and reliable method to measure changes in the extent of phosphorylation is lacking. Here, we present a method to quantitate the changes in phosphorylation occurring in a protein in response to a stimulus. The method consists of three steps: (i) enzymatic digestion in H(2)16O or isotopically enriched H(2)18O to label individual pools of differentially phosphorylated proteins; (ii) affinity selection of phosphopeptides from the combined digests by immobilized metal-affinity chromatography; and (iii) dephosphorylation with alkaline phosphatase to allow for quantitation of changes of phosphorylation by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. We applied this strategy to the analysis of the yeast nitrogen permease reactivator protein kinase involved in the target of rapamycin signaling pathway. Alteration in the extent of phosphorylation at Ser-353 and Ser-357 could be easily assessed and quantitated both in wild-type yeast cells treated with rapamycin and in cells lacking the SIT4 phosphatase responsible for dephosphorylating nitrogen permease reactivator protein. The method described here is simple and allows quantitation of relative changes in the level of phosphorylation in signaling proteins, thus yielding information critical for understanding the regulation of complex protein phosphorylation cascades. PMID- 12540833 TI - Organic hydroperoxide resistance gene encodes a thiol-dependent peroxidase. AB - ohr (organic hydroperoxide resistance gene) is present in several species of bacteria, and its deletion renders cells specifically sensitive to organic peroxides. The goal of this work was to determine the biochemical function of Ohr from Xylella fastidiosa. All of the Ohr homologues possess two cysteine residues, one of them located in a VCP motif, which is also present in all of the proteins from the peroxiredoxin family. Therefore, we have investigated whether Ohr possesses thiol-dependent peroxidase activity. The ohr gene from X. fastidiosa was expressed in Escherichia coli, and the recombinant Ohr decomposed hydroperoxides in a dithiothreitol-dependent manner. Ohr was about twenty times more efficient to remove organic hydroperoxides than to remove H(2)O(2). This result is consistent with the organic hydroperoxide sensitivity of Delta ohr strains. The dependence of Ohr on thiol compounds was ascertained by glutamine synthetase protection assays. Approximately two thiol equivalents were consumed per peroxide removed indicating that Ohr catalyzes the following reaction: 2RSH + ROOH --> RSSR + ROH + H(2)O. Pretreatment of Ohr with N-ethyl maleimide and substitution of cysteine residues by serines inhibited this peroxidase activity indicating that both of the Ohr cysteines are important to the decomposition of peroxides. C125S still had a residual enzymatic activity indicating that Cys-61 is directly involved in peroxide removal. Monothiol compounds do not support the peroxidase activity of Ohr as well as thioredoxin from Saccharomyces cerevisiae and from Spirulina. Interestingly, dithiothreitol and dyhydrolipoic acid, which possess two sulfhydryl groups, do support the peroxidase activity of Ohr. Taken together our results unequivocally demonstrated that Ohr is a thiol-dependent peroxidase. PMID- 12540832 TI - Photochemical gating of heterologous ion channels: remote control over genetically designated populations of neurons. AB - Heterologous proteins capable of transducing physical or chemical stimuli into electrical signals can be used to control the function of excitable cells in intact tissues or organisms. Restricted genetically to circumscribed populations of cellular targets, these selectively addressable sources of depolarizing current can supply distributed inputs to neural circuits, stimulate secretion, or regulate force and motility. In an initial demonstration of this principle, we have used elements of a G protein coupled signaling system, the phototransduction cascade of the fruit fly, to sensitize generalist vertebrate neurons to light [Zemelman, B. V., Lee, G. A., Ng, M. & Miesenbock, G. (2002) Neuron 33, 15-22]. We now describe the use of ectopically expressed ligand-gated ion channels as transducers of optical or pharmacological stimuli. When either the capsaicin receptor, TRPV1, the menthol receptor, TRPM8, or the ionotropic purinergic receptor P2X(2) was introduced into hippocampal neurons, the cells responded to pulsed applications of agonist with characteristic sequences of depolarization, spiking, and repolarization. Responses required cognate matches between receptor and agonist, peaked at firing frequencies of approximately 40 Hz, initiated and terminated rapidly, and did not attenuate. Precise dose-response relationships allowed current amplitudes and firing frequencies to be tuned by varying the concentration of ligand. Agonist could be administered either pharmacologically or, in the cases of TRPV1 and P2X(2), optically, through photorelease of the active compounds from the respective "caged" precursors, 4,5-dimethoxy-2 nitrobenzyl-capsaicin and P(3)-[1-(4,5-dimethoxy-2-nitrophenyl)ethyl]-ATP. PMID- 12540834 TI - A single amino acid difference between alpha and beta Ca2+/calmodulin-dependent protein kinase kinase dictates sensitivity to the specific inhibitor, STO-609. AB - We recently developed STO-609, a selective inhibitor of Ca(2+)/calmodulin dependent protein kinase kinase (CaM-KK), and we demonstrated that CaM-KK beta is more sensitive to STO-609 than the CaM-KK alpha isoform (Tokumitsu H., Inuzuka H., Ishikawa Y., Ikeda M., Saji I., and Kobayashi R. (2002) J. Biol. Chem. 277, 15813-15818). By using catalytic chimera and point mutants of both isoforms, we demonstrated that Val(269) in CaM-KK beta/Leu(233) in CaM-KK alpha confers a distinct sensitivity ( approximately 10-fold) to STO-609 on CaM-KK isoforms. Various mutations of Val(269) in CaM-KK beta indicate that substitution by hydrophobic residues with bulky side chains significantly decreases drug sensitivity and that the V269F mutant is the most effective drug-resistant enzyme ( approximately 80-fold higher IC(50) value). These findings are consistent with a result obtained with a full-length mutant expressed in COS-7 cells. Furthermore, suppression of CaM-KK-mediated CaM-KIV activation in transfected HeLa cells by STO-609 treatment was completely abolished by the co-expression of the CaM-KK beta V269F mutant. Based on the results that the distinct sensitivity of CaM-KK isoforms to STO-609 is because of a single amino acid substitution (Val/Leu) in the ATP-binding pocket, we have generated an STO-609-resistant CaM KK mutant, which might be useful for validating the pharmacological effects and specificity of STO-609 in vivo. PMID- 12540835 TI - Catalytic mechanism of Escherichia coli isopentenyl diphosphate isomerase involves Cys-67, Glu-116, and Tyr-104 as suggested by crystal structures of complexes with transition state analogues and irreversible inhibitors. AB - Isopentenyl diphosphate (IPP):dimethylallyl diphosphate (DMAPP) isomerase is a key enzyme in the biosynthesis of isoprenoids. The reaction involves protonation and deprotonation of the isoprenoid unit and proceeds through a carbocationic transition state. Analysis of the crystal structures (2 A) of complexes of Escherichia coli IPP.DMAPPs isomerase with a transition state analogue (N,N dimethyl-2-amino-1-ethyl diphosphate) and a covalently attached irreversible inhibitor (3,4-epoxy-3-methyl-1-butyl diphosphate) indicates that Glu-116, Tyr 104, and Cys-67 are involved in the antarafacial addition/elimination of protons during isomerization. This work provides a new perspective about the mechanism of the reaction. PMID- 12540836 TI - The NO pathway acts late during the fertilization response in sea urchin eggs. AB - Both the inositol 1,4,5-trisphosphate (InsP(3)) and ryanodine receptor pathways contribute to the Ca(2+) transient at fertilization in sea urchin eggs. To date, the precise contribution of each pathway has been difficult to ascertain. Evidence has accumulated to suggest that the InsP(3) receptor pathway has a primary role in causing Ca(2+) release and egg activation. However, this was recently called into question by a report implicating NO as the primary egg activator. In the present study we pursue the hypothesis that NO is a primary egg activator in sea urchin eggs and build on previous findings that an NO/cGMP/cyclic ADP-ribose (cADPR) pathway is active at fertilization in sea urchin eggs to define its role. Using a fluorescence indicator of NO levels, we have measured both NO and Ca(2+) at fertilization and establish that NO levels rise after, not before, the Ca(2+) wave is initiated and that this rise is Ca(2+) dependent. By inhibiting the increase in NO at fertilization, we find not that the Ca(2+) transient is abolished but that the duration of the transient is significantly reduced. The latency and rise time of the transient are unaffected. This effect is mirrored by the inhibition of cGMP and cADPR signaling in sea urchin eggs at fertilization. We establish that cADPR is generated at fertilization, at a time comparable to the time of the rise in NO levels. We conclude that NO is unlikely to be a primary egg activator but, rather, acts after the initiation of the Ca(2+) wave to regulate the duration of the fertilization Ca(2+) transient. PMID- 12540837 TI - Identification by mutagenesis of conserved arginine and glutamate residues in the C-terminal domain of rat liver carnitine palmitoyltransferase I that are important for catalytic activity and malonyl-CoA sensitivity. AB - Carnitine palmitoyltransferase I (CPTI) catalyzes the conversion of long chain fatty acyl-CoAs to acylcarnitines in the presence of l-carnitine. To determine the role of the conserved glutamate residue, Glu-603, on catalysis and malonyl CoA sensitivity, we separately changed the residue to alanine, histidine, glutamine, and aspartate. Substitution of Glu-603 with alanine or histidine resulted in complete loss of L-CPTI activity. A change of Glu-603 to glutamine caused a significant decrease in catalytic activity and malonyl-CoA sensitivity. Substitution of Glu-603 with aspartate, a negatively charged amino acid with only one methyl group less than the glutamate residue in the wild type enzyme, resulted in partial loss in CPTI activity and a 15-fold decrease in malonyl-CoA sensitivity. The mutant L-CPTI with a replacement of the conserved Arg-601 or Arg 606 with alanine also showed over 40-fold decrease in malonyl-CoA sensitivity, suggesting that these two conserved residues may be important for substrate and inhibitor binding. Since a conservative substitution of Glu-603 to aspartate or glutamine resulted in partial loss of activity and malonyl-CoA sensitivity, it further suggests that the negative charge and the longer side chain of glutamate are essential for catalysis and malonyl-CoA sensitivity. We predict that this region of L-CPTI spanning these conserved C-terminal residues may be the region of the protein involved in binding the CoA moiety of palmitoyl-CoA and malonyl CoA and/or the putative low affinity acyl-CoA/malonyl-CoA binding site. PMID- 12540838 TI - Persistent ERK phosphorylation negatively regulates cAMP response element-binding protein (CREB) activity via recruitment of CREB-binding protein to pp90RSK. AB - Compound 5 (Cpd 5) or 2-(2-mercaptoethanol)-3-methyl-1,4-naphthoquinone, is an inhibitor of protein phosphatase Cdc25A and causes persistent activation of extracellular signal-regulated kinase (ERK) and cell growth inhibition. To study the mechanism(s) by which persistent ERK phosphorylation might induce cell growth inhibition, we used Cpd 5 as a tool to examine its effects on the activity of CREB (cAMP response element-binding protein) transcription factor in Hep3B human hepatoma cells. We found that CREB activity, including its DNA binding ability and phosphorylation on residue Ser-133, was strongly inhibited by Cpd 5, followed by suppression of CRE-mediated transcription of cyclin D1 and Bcl-2 genes. Cpd 5 mediated suppression of CREB phosphorylation and transcriptional activity was antagonized by mitogen-activated protein kinase kinase inhibitors PD 98059 and U 0126, implying that this inhibition of CREB activity was regulated at least in part by the ERK pathway. The phosphorylation of ribosomal S6 kinase (pp90(RSK)), a CREB kinase in response to mitogen stimulation, was also found to be inhibited by Cpd 5 action. This inhibition of pp90(RSK) phosphorylation is likely the result of its increased association with CREB-binding protein (CBP), which subsequently caused inhibition of CREB phosphorylation and activity. To support the hypothesis that Cpd 5 effects on Cdc25A inhibition with subsequent ERK activation could cause CREB inhibition, we examined the effects of Cdc25A inhibition without the use of Cpd 5. Hep3B cells were transfected with C430S Cdc25A mutant, and ERK was found to be phosphorylated in a constitutively activated manner, which was accompanied by decreased CREB phosphorylation and increased recruitment of CBP to pp90(RSK). These data provide evidence that CBP.RSK complex formation in response to persistent ERK phosphorylation by Cpd 5 down-regulates CREB activity, leading to inhibition of both cAMP response element mediated gene expression and cell growth. PMID- 12540839 TI - YC-1 facilitates release of the proximal His residue in the NO and CO complexes of soluble guanylate cyclase. AB - The benzylindazole compound YC-1 has been shown to activate soluble guanylate cyclase by increasing the sensitivity toward NO and CO. Here we report the action of YC-1 on the coordination of CO- and NO-hemes in the enzyme and correlate the events with the activation of enzyme catalysis. A single YC-1-binding site on the heterodimeric enzyme was identified by equilibrium dialysis. To explore the affect of YC-1 on the NO-heme coordination, the six-coordinate NO complex of the enzyme was stabilized by dibromodeuteroheme substitution. Using the dibromodeuteroheme enzyme, YC-1 converted the six-coordinate NO-heme to a five coordinate NO-heme with a characteristic EPR signal that differed from that in the absence of YC-1. These results revealed that YC-1 facilitated cleavage of the proximal His-iron bond and caused geometrical distortion of the five-coordinate NO-heme. Resonance Raman studies demonstrated the presence of two iron-CO stretch modes at 488 and 521 cm(-1) specific to the YC-1-bound CO complex of the native enzyme. Together with the infrared C-O stretching measurements, we assigned the 488-cm(-1) band to the iron-CO stretch of a six-coordinate CO-heme and the 521 cm(-1) band to the iron-CO stretch of a five-coordinate CO-heme. These results indicate that YC-1 stimulates enzyme activity by weakening or cleaving the proximal His-iron bond in the CO complex as well as the NO complex. PMID- 12540840 TI - Inhibition of human SERCA3 by PL/IM430. Molecular analysis of the interaction. AB - The monoclonal antibody PL/IM430 has previously been reported to uncouple Ca(2+) transport from ATP hydrolysis in platelet membranes (Hack, N., Wilkinson, J. M., and Crawford, N. (1988) Biochem. J. 250, 355-361). More recently, we have demonstrated that this antibody is specific for human SERCA3 (Poch, E., Leach, S., Snape, S., Cacic, T., MacLennan, D. H., and Lytton, J. (1998) Am. J. Physiol. 275, C1449-C1458). In this paper, we have extended the analysis of the PL/IM430 SERCA3 interaction. Using HEK293 cells to express human SERCA3a, we were able to measure both ATP-mediated, oxalate-dependent (45)Ca(2+) uptake and Ca(2+) dependent ATP hydrolysis activities due exclusively to SERCA3. Treatment with PL/IM430 inhibited both activities almost identically, with a maximal inhibition of 81 and 73% and a half-maximal concentration of 8.3 and 5.9 microg/ml, for Ca(2+) uptake and ATP hydrolysis, respectively. We conclude that PL/IM430 does inhibit SERCA3 activity but does not uncouple Ca(2+) transport from ATP hydrolysis. Using a combination of partial proteolysis, GST fusion protein expression, and mutation of residues that differ between rat and human SERCA3, we have identified human SERCA3 amino acids Pro(8) and Glu(192) as essential to forming the PL/IM430 epitope. PL/IM430 thus recognizes a linearly noncontiguous set of amino acids within the actuator domain of human SERCA3. We propose that PL/IM430 inhibits SERCA3 activity by sterically preventing movement of the actuator domain into a catalytically critical position in the E2 conformation of the enzyme. PMID- 12540841 TI - Promyelocytic leukemia protein sensitizes tumor necrosis factor alpha-induced apoptosis by inhibiting the NF-kappaB survival pathway. AB - The promyelocytic leukemia protein (PML) is a growth/tumor suppressor essential for induction of apoptosis by diverse apoptotic stimuli. The mechanism by which PML regulates cell death remains unclear. In this study we found that ectopic expression of PML potentiates cell death by apoptosis in the tumor necrosis factor alpha (TNFalpha)-resistant cell line U2OS and other cell lines. Treatment with TNFalpha significantly sensitized these cells to apoptosis in a p53 independent manner. PML/TNFalpha-induced cell death is associated with DNA fragmentation, activation of caspase-3, -7, and -8, and degradation of DNA fragmentation factor/inhibitor of CAD. PML/TNFalpha-induced cell death could be blocked by the caspase-8 inhibitors CrmA and c-FLIP but not by Bcl-2. These findings indicate that this cell death event is initiated through the death receptor-dependent apoptosis pathway. PML is a transcriptional repressor of NF kappaB by interacting with RelA/p65 and prevents its binding to the cognate enhancer through the C terminus. Coimmunoprecipitation and double-color immunofluorescence staining demonstrated that PML physically interacts with RelA/p65 in vivo and the two proteins colocalized at the endogenous levels. Overexpression of NF-kappaB rescued cell death induced by PML/TNFalpha. Furthermore, PML(-/-) mouse embryo fibroblasts are more resistant to TNFalpha induced apoptosis. Together this study defines a novel mechanism by which PML induces apoptosis through repression of the NF-kappaB survival pathway. PMID- 12540842 TI - STAP-2/BKS, an adaptor/docking protein, modulates STAT3 activation in acute-phase response through its YXXQ motif. AB - As a c-fms-interacting protein, we cloned a novel adaptor molecule, signal transducing adaptor protein-2 (STAP-2), which contains pleckstrin homology- and Src homology 2-like (PH and SRC) domains and a proline-rich region. STAP-2 is structurally related to STAP-1/BRDG1 (BCR downstream signaling-1), which we had cloned previously from hematopoietic stem cells. STAP-2 is a murine homologue of a recently identified adaptor molecule, BKS, a substrate of BRK tyrosine kinase. STAP-2 was tyrosine-phosphorylated and translocated to the plasma membrane in response to epidermal growth factor when overexpressed in fibroblastic cells. To define the function of STAP-2, we generated mice lacking the STAP-2 gene. STAP-2 mRNA was strongly induced in the liver in response to lipopolysaccharide and in isolated hepatocytes in response to interleukin-6. In the STAP-2(-/-) hepatocytes, the interleukin-6-induced expression of acute-phase (AP) genes and the tyrosine-phosphorylation level of STAT3 were reduced specifically at the late phase (6-24 h) of the response. These data indicate that STAP-2 plays a regulatory role in the AP response in systemic inflammation. STAP-2 contains a YXXQ motif in the C-terminal region that is a potential STAT3-binding site. Overexpression of wild-type STAP-2, but not of mutants lacking this motif, enhanced the AP response element reporter activity and an AP protein production. These data suggest that STAP-2 is a new class of adaptor molecule that modulates STAT3 activity through its YXXQ motif. PMID- 12540843 TI - Hierarchical affinities and a bipartite interaction model for estrogen receptor isoforms and full-length steroid receptor coactivator (SRC/p160) family members. AB - Nuclear receptor (NR)-mediated transcription is driven by dynamic multiprotein coactivator complexes, the composition of which is thought to determine the biological activity of NRs at specific promoters. The extent to which NRs discriminate between a spectrum of potential binding partners is intuitively a function of the inherent affinities of these individual interactions. Using real time interaction analysis with BIAcore, we evaluated the affinities and kinetics of the interactions of full-length members of the SRC/p160 coactivator family with estrogen receptor alpha (ER alpha) and ER beta bound to a variety of ligands. We substantiate that 17beta-estradiol enhances the affinity of ER SRC/p160 interactions, whereas 4(OH)-tamoxifen, raloxifene, and ICI-182,780 inhibit these interactions. We show that a well defined, ER isoform-specific hierarchy governs the association of liganded ERs with full length SRC/p160 family members. Moreover, our data indicate that the interaction affinities of the full-length SRC/p160s with ERs are significantly higher then those of the NR interaction domains of the same coactivators, indicating that portions of coactivator molecules distinct from NR interaction domains might participate in receptor-coactivator complex formation. Finally, the interaction kinetics of SRC/p160s with ERs are consistent with a bipartite model, involving initial rapid formation of an unstable intermediate complex, and a subsequent slower reaction leading to its stabilization. We interpret our results as evidence that hierarchical coactivator interaction affinities are an important source of diversity in NR-mediated signaling and that the complexity of receptor coactivator cross-talk might be best understood in the context of full-length molecules. PMID- 12540844 TI - Role of the extracellular and cytoplasmic domains of CD44 in the rolling interaction of lymphoid cells with hyaluronan under physiologic flow. AB - CD44 can function as an adhesion receptor that mediates leukocyte rolling on hyaluronan (HA). To study the contributions of different domains of the standard isoform of CD44 to cell rolling, a CD44-negative mouse T lymphoma AKR1 was transfected with wild type (WT) or mutated cDNA constructs. A parallel flow chamber was used to study the rolling behavior of CD44 transfectants on immobilized HA. For CD44WT transfectants, the fraction of cells that rolled and the rolling velocity was inversely proportional to the amount of cell surface CD44. When the cytoplasmic domain distal to Gly(305) or sequences that serve as binding sites for cytoskeletal linker proteins, were deleted or replaced with foreign sequences, no significant changes in the rolling behavior of mutant cells, compared with the transfectant expressing CD44WT, were observed. Transfectants lacking 64 amino acids of the cytoplasmic tail distal to Cys(295) adhered to HA but showed enhanced rolling at low shear forces. When 83 amino acids from the "non-conserved" membrane-proximal region of the CD44 extracellular domain were deleted, cells adhered firmly to the HA substrate and did not roll at any fluid shear force tested. Unlike wild type cells that exhibited a nearly homogeneous distribution of CD44 on a smooth cell surface, cells expressing the non-conserved region deletion mutant accumulated CD44 in membrane protrusions. Disruption of the actin cytoskeleton with cytochalasin B precluded the formation of membrane protrusions, however, treated cells still adhered firmly to HA and did not roll. We conclude that interaction between the cytoplasmic domain of CD44 and the cytoskeleton is not required for cell rolling on immobilized ligand. The strong effect of deletion of the non-conserved region of the extracellular domain argues for a critical role of this region in CD44-dependent rolling and adhesion interactions with HA under flow. PMID- 12540845 TI - Characterization and implications of Ca2+ binding to pectate lyase C. AB - Ca(2+) is essential for in vitro activity of Erwinia chrysanthemi pectate lyase C (PelC). Crystallographic analyses of 11 PelC-Ca(2+) complexes, formed at pH 4.5, 9.5, and 11.2 under varying Ca(2+) concentrations, have been solved and refined at a resolution of 2.2 A. The Ca(2+) site represents a new motif for Ca(2+), consisting primarily of beta-turns and beta-strands. The principal differences between PelC and the PelC-Ca(2+) structures at all pH values are the side-chain conformations of Asp-129 and Glu-166 as well as the occupancies of four water molecules. According to calculations of pK(a) values, the presence of Ca(2+) and associated structural changes lower the pK(a) of Arg-218, the amino acid responsible for proton abstraction during catalysis. The Ca(2+) affinity for PelC is weak, as the K(d) was estimated to be 0.132 (+/-0.004) mm at pH 9.5, 1.09 (+/ 0.29) mm at pH 11.2, and 5.84 (+/-0.41) mm at pH 4.5 from x-ray diffraction studies and 0.133 (+/-0.045) mm at pH 9.5 from intrinsic tryptophan fluorescence measurements. Given the pH dependence of Ca(2+) affinity, PelC activity at pH 4.5 has been reexamined. At saturating Ca(2+) concentrations, PelC activity increases 10-fold at pH 4.5 but is less than 1% of maximal activity at pH 9.5. Taken together, the studies suggest that the primary Ca(2+) ion in PelC has multiple functions. PMID- 12540846 TI - The chemoattractant receptor-like protein C5L2 binds the C3a des-Arg77/acylation stimulating protein. AB - The orphan receptor C5L2 has recently been described as a high affinity binding protein for complement fragments C5a and C3a that, unlike the previously described C5a receptor (CD88), couples only weakly to G(i)-like G proteins (Cain, S. A., and Monk, P. N. (2002) J. Biol. Chem. 277, 7165-7169). Here we demonstrate that C5L2 binds the metabolites of C4a and C3a, C4a des-Arg(77), and C3a des Arg(77) (also known as the acylation-stimulating protein or ASP) at a site distinct from the C5a binding site. The binding of these metabolites to C5L2 does not stimulate the degranulation of transfected rat basophilic leukemia cells either through endogenous rat G proteins or when co-transfected with human G(alpha 16). C3a des-Arg(77)/ASP and C3a can potently stimulate triglyceride synthesis in human skin fibroblasts and 3T3-L1 preadipocytes. Here we show that both cell types and human adipose tissue express C5L2 mRNA and that the human fibroblasts express C5L2 protein at the cell surface. This is the first demonstration of the expression of C5L2 in cells that bind and respond to C3a des Arg(77)/ASP and C3a. Thus C5L2, a promiscuous complement fragment-binding protein with a high affinity site that binds C3a des-Arg(77)/ASP, may mediate the acylation-stimulating properties of this peptide. PMID- 12540847 TI - Collagen triple helix formation can be nucleated at either end. AB - The directional dependence of folding rates for rod-like macromolecules such as parallel alpha-helical coiled-coils, DNA double-helices, and collagen triple helices is largely unexplored. This is mainly due to technical difficulties in measuring rates in different directions. Folding of collagens is nucleated by trimeric non-collagenous domains. These are usually located at the COOH terminus, suggesting that triple helix folding proceeds from the COOH to the NH(2) terminus. Evidence is presented here that effective nucleation is possible at both ends of the collagen-like peptide (Gly-Pro-Pro)(10), using designed proteins in which this peptide is fused either NH(2)- or COOH-terminal to a nucleation domain, either T4-phage foldon or the disulfide knot of type III collagen. The location of the nucleation domain influences triple-helical stability, which might be explained by differences in the linker sequences and the presence or absence of repulsive charges at the carboxyl-terminal end of the triple helix. Triple helical folding rates are found to be independent of the site of nucleation and consistent with cis-trans isomerization being the rate-limiting step. PMID- 12540848 TI - Single-stranded DNA induces ataxia telangiectasia mutant (ATM)/p53-dependent DNA damage and apoptotic signals. AB - Single-stranded DNA has been speculated to be the initial signal in the DNA damage signaling pathway. We showed that introduction of single-stranded DNA with diverse sequences into mammalian cells induced DNA damage as well as apoptosis signals. Like DNA damaging agents, single-stranded DNA up-regulated p53 and activated the nuclear kinase ataxia telangiectasia mutant (ATM) as evidenced by phosphorylation of histone 2AX, an endogenous ATM substrate. Single-stranded DNA also triggered apoptosis as evidenced by the formation of caspase-dependent chromosomal DNA strand breaks, cytochrome c release, and increase in reactive oxygen species production. Moreover, single-stranded DNA-induced apoptosis was reduced significantly in p53 null cells and in cells treated with ATM small interfering RNA. These results suggest that single-stranded DNA may act upstream of ATM/p53 in DNA damage signaling. PMID- 12540849 TI - Kinetic stabilization of Bacillus licheniformis alpha-amylase through introduction of hydrophobic residues at the surface. AB - It is generally assumed that in proteins hydrophobic residues are not favorable at solvent-exposed sites, and that amino acid substitutions on the surface have little effect on protein thermostability. Contrary to these assumptions, we have identified hyperthermostable variants of Bacillus licheniformis alpha-amylase (BLA) that result from the incorporation of hydrophobic residues at the surface. Under highly destabilizing conditions, a variant combining five stabilizing mutations unfolds 32 times more slowly and at a temperature 13 degrees C higher than the wild-type. Crystal structure analysis at 1.7 A resolution suggests that stabilization is achieved through (a) extension of the concept of increased hydrophobic packing, usually applied to cavities, to surface indentations, (b) introduction of favorable aromatic-aromatic interactions on the surface, (c) specific stabilization of intrinsic metal binding sites, and (d) stabilization of a beta-sheet by introducing a residue with high beta-sheet forming propensity. All mutated residues are involved in forming complex, cooperative interaction networks that extend from the interior of the protein to its surface and which may therefore constitute "weak points" where BLA unfolding is initiated. This might explain the unexpectedly large effect induced by some of the substitutions on the kinetic stability of BLA. Our study shows that substantial protein stabilization can be achieved by stabilizing surface positions that participate in underlying cooperatively formed substructures. At such positions, even the apparently thermodynamically unfavorable introduction of hydrophobic residues should be explored. PMID- 12540850 TI - Hepatitis C virus internal ribosome entry site-mediated translation is stimulated by specific interaction of independent regions of human La autoantigen. AB - The human La autoantigen has been shown to interact with the internal ribosome entry site (IRES) of hepatitis C virus (HCV) in vitro. Using a yeast three-hybrid system, we demonstrated that, in addition to full-length La protein, both N- and C-terminal halves were able to interact with HCV IRES in vivo. The exogenous addition of purified full-length and truncated La proteins in rabbit reticulocyte lysate showed dose-dependent stimulation of HCV IRES-mediated translation. However, an additive effect was achieved adding the terminal halves together in the reaction, suggesting that both might play critical roles in achieving full stimulatory activity of the full-length La protein. Using computational analysis, three-dimensional structures of the RNA recognition motifs (RRM) of the La protein were independently modeled. Of the three putative RRMs, RRM2 was predicted to have a good binding pocket for the interaction with the HCV IRES around the GCAC motif near the initiator AUG and RRM3 binds perhaps in a different location. This observation was further investigated by the filter binding and toe-printing assays. The results presented here strongly suggest that both the N- and C-terminal halves can interact independently with the HCV IRES and are involved in stimulating internal initiation of translation. PMID- 12540851 TI - Haematopoietic stem cells retain long-term repopulating activity and multipotency in the absence of stem-cell leukaemia SCL/tal-1 gene. AB - The production of blood cells is sustained throughout the lifetime of an individual by haematopoietic stem cells (HSCs). Specification of HSCs from mesoderm during embryonic development requires the stem cell leukaemia SCL/tal-1 gene product. Forced expression of SCL/tal-1 strongly induces blood formation in embryos, indicating that this gene has a dominant role in commitment to haematopoiesis. In the adult haematopoietic system, expression of SCL/tal-1 is enriched in HSCs and multipotent progenitors, and in erythroid and megakaryocytic lineages, consistent with roles for this factor in adult haematopoiesis. Here we assess by conditional gene targeting whether SCL/tal-1 is required continuously for the identity and function of HSCs. We find that SCL/tal-1 is dispensable for HSC engraftment, self-renewal and differentiation into myeloid and lymphoid lineages; however, the proper differentiation of erythroid and megakaryocytic precursors is dependent on SCL/tal-1. Thus, SCL/tal-1 is essential for the genesis of HSCs, but its continued expression is not essential for HSC functions. These findings contrast with lineage choice mechanisms, in which the identity of haematopoietic lineages requires continuous transcription factor expression. PMID- 12540852 TI - Effects of household dynamics on resource consumption and biodiversity. AB - Human population size and growth rate are often considered important drivers of biodiversity loss, whereas household dynamics are usually neglected. Aggregate demographic statistics may mask substantial changes in the size and number of households, and their effects on biodiversity. Household dynamics influence per capita consumption and thus biodiversity through, for example, consumption of wood for fuel, habitat alteration for home building and associated activities, and greenhouse gas emissions. Here we report that growth in household numbers globally, and particularly in countries with biodiversity hotspots (areas rich in endemic species and threatened by human activities), was more rapid than aggregate population growth between 1985 and 2000. Even when population size declined, the number of households increased substantially. Had the average household size (that is, the number of occupants) remained static, there would have been 155 million fewer households in hotspot countries in 2000. Reduction in average household size alone will add a projected 233 million additional households to hotspot countries during the period 2000-15. Rapid increase in household numbers, often manifested as urban sprawl, and resultant higher per capita resource consumption in smaller households pose serious challenges to biodiversity conservation. PMID- 12540853 TI - Fidelity in planar cell polarity signalling. AB - The polarity of Drosophila wing hairs displays remarkable fidelity. Each of the approximately 30,000 wing epithelial cells constructs an actin-rich prehair that protrudes from its distal vertex and points distally. The distal location and orientation of the hairs is virtually error free, thus forming a nearly perfect parallel array. This process is controlled by the planar cell polarity signalling pathway. Here we show that interaction between two tiers of the planar cell polarity signalling mechanism results in the observed high fidelity. The first tier, mediated by the cadherin Fat, dictates global orientation by transducing a directional signal to individual cells. The second tier, orchestrated by the 7 pass transmembrane receptor Frizzled, aligns each cell's polarity with that of its neighbours through the action of an intercellular feedback loop, enabling polarity to propagate from cell to cell. We show that all cells need not respond correctly to the presumably subtle signal transmitted by Fat. Subsequent action of the Frizzled feedback loop is sufficient to align all the cells cooperatively. This economical system is therefore highly robust, and produces virtually error free arrays. PMID- 12540854 TI - Crystal structure of the human angiotensin-converting enzyme-lisinopril complex. AB - Angiotensin-converting enzyme (ACE) has a critical role in cardiovascular function by cleaving the carboxy terminal His-Leu dipeptide from angiotensin I to produce a potent vasopressor octapeptide, angiotensin II. Inhibitors of ACE are a first line of therapy for hypertension, heart failure, myocardial infarction and diabetic nephropathy. Notably, these inhibitors were developed without knowledge of the structure of human ACE, but were instead designed on the basis of an assumed mechanistic homology with carboxypeptidase A. Here we present the X-ray structure of human testicular ACE and its complex with one of the most widely used inhibitors, lisinopril (N2-[(S)-1-carboxy-3-phenylpropyl]-L-lysyl-L-proline; also known as Prinivil or Zestril), at 2.0 A resolution. Analysis of the three dimensional structure of ACE shows that it bears little similarity to that of carboxypeptidase A, but instead resembles neurolysin and Pyrococcus furiosus carboxypeptidase--zinc metallopeptidases with no detectable sequence similarity to ACE. The structure provides an opportunity to design domain-selective ACE inhibitors that may exhibit new pharmacological profiles. PMID- 12540855 TI - Structure and catalytic mechanism of the human histone methyltransferase SET7/9. AB - Acetylation, phosphorylation and methylation of the amino-terminal tails of histones are thought to be involved in the regulation of chromatin structure and function. With just one exception, the enzymes identified in the methylation of specific lysine residues on histones (histone methyltransferases) belong to the SET family. The high-resolution crystal structure of a ternary complex of human SET7/9 with a histone peptide and cofactor reveals that the peptide substrate and cofactor bind on opposite surfaces of the enzyme. The target lysine accesses the active site of the enzyme and the S-adenosyl-l-methionine (AdoMet) cofactor by inserting its side chain into a narrow channel that runs through the enzyme, connecting the two surfaces. Here we show from the structure and from solution studies that SET7/9, unlike most other SET proteins, is exclusively a mono methylase. The structure indicates the molecular basis of the specificity of the enzyme for the histone target, and allows us to propose a model for the methylation reaction that accounts for the role of many of the residues that are invariant across the SET family. PMID- 12540856 TI - Telomere dysfunction and Atm deficiency compromises organ homeostasis and accelerates ageing. AB - Ataxia-telangiectasia (A-T) results from the loss of ataxia-telangiectasia mutated (Atm) function and is characterized by accelerated telomere loss, genomic instability, progressive neurological degeneration, premature ageing and increased neoplasia incidence. Here we evaluate the functional interaction of Atm and telomeres in vivo. We examined the impact of Atm deficiency as a function of progressive telomere attrition at both the cellular and whole-organism level in mice doubly null for Atm and the telomerase RNA component (Terc). These compound mutants showed increased telomere erosion and genomic instability, yet they experienced a substantial elimination of T-cell lymphomas associated with Atm deficiency. A generalized proliferation defect was evident in all cell types and tissues examined, and this defect extended to tissue stem/progenitor cell compartments, thereby providing a basis for progressive multi-organ system compromise, accelerated ageing and premature death. We show that Atm deficiency and telomere dysfunction act together to impair cellular and whole-organism viability, thus supporting the view that aspects of A-T pathophysiology are linked to the functional state of telomeres and its adverse effects on stem/progenitor cell reserves. PMID- 12540857 TI - Nanotech is not so scary. PMID- 12540858 TI - Fair play for trial balloons. PMID- 12540859 TI - Failed mission launch prompts ESA to reconsider comet target. PMID- 12540860 TI - Report backs military axis on space. PMID- 12540861 TI - Mozambique prime minister tipped for global health post. PMID- 12540862 TI - Bushfires annihilate Australian observatory. PMID- 12540863 TI - Resignations rock mathematics institute. PMID- 12540864 TI - Researcher uncovers truth behind wartime security slur. PMID- 12540865 TI - Transgenic salmon still out in the cold in United States. PMID- 12540866 TI - Tokyo deans plan flexible future for students. PMID- 12540867 TI - Second cancer case halts gene-therapy trials. PMID- 12540868 TI - Meeting aims to find brain's benchmarks for beauty. PMID- 12540870 TI - Balloon-based astronomy: up, up and away. PMID- 12540871 TI - DNA: Beyond the double helix. PMID- 12540872 TI - Travel grants available for genetics congress. PMID- 12540873 TI - Spiritual link is part of traditional knowledge. PMID- 12540874 TI - A scarlet letter or a red herring? PMID- 12540875 TI - Is a scientific boycott ever justified? PMID- 12540881 TI - Mathematical oncology: cancer summed up. PMID- 12540882 TI - Palaeontology: Dinosaurs take to the air. PMID- 12540883 TI - Global change: Monsoon linkages. PMID- 12540885 TI - Thermal physics: Heat in one dimension. PMID- 12540884 TI - Neurobiology: Fear thou not. PMID- 12540886 TI - Inflammation: A nervous connection. PMID- 12540887 TI - Astronomy: Feeding the first quasars. PMID- 12540890 TI - Flight performance: Frigatebirds ride high on thermals. PMID- 12540891 TI - Pollinator attraction: Crab-spiders manipulate flower signals. PMID- 12540893 TI - Spectral signature of cosmological infall of gas around the first quasars. AB - Recent observations have shown that, only a billion years after the Big Bang, the Universe was already lit up by bright quasars fuelled by the infall of gas onto supermassive black holes. The masses of these early black holes are inferred from their luminosities to be >10(9) solar masses (M(O)), which is a difficult theoretical challenge to explain. Like nearby quasars, the early objects could have formed in the central cores of massive host galaxies. The formation of these hosts could be explained if, like local large galaxies, they were assembled gravitationally inside massive (> 10(12) M(O)) haloes of dark matter. There has hitherto been no observational evidence for the presence of these massive hosts or their surrounding haloes. Here we show that the cosmic gas surrounding each halo must respond to its strong gravitational pull, where absorption by the infalling hydrogen produces a distinct spectral signature. That signature can be seen in recent data. PMID- 12540892 TI - Four-winged dinosaurs from China. AB - Although the dinosaurian hypothesis of bird origins is widely accepted, debate remains about how the ancestor of birds first learned to fly. Here we provide new evidence suggesting that basal dromaeosaurid dinosaurs were four-winged animals and probably could glide, representing an intermediate stage towards the active, flapping-flight stage. The new discovery conforms to the predictions of early hypotheses that proavians passed through a tetrapteryx stage. PMID- 12540894 TI - Experimental extraction of an entangled photon pair from two identically decohered pairs. AB - Entanglement is considered to be one of the most important resources in quantum information processing schemes, including teleportation, dense coding and entanglement-based quantum key distribution. Because entanglement cannot be generated by classical communication between distant parties, distribution of entangled particles between them is necessary. During the distribution process, entanglement between the particles is degraded by the decoherence and dissipation processes that result from unavoidable coupling with the environment. Entanglement distillation and concentration schemes are therefore needed to extract pairs with a higher degree of entanglement from these less-entangled pairs; this is accomplished using local operations and classical communication. Here we report an experimental demonstration of extraction of a polarization entangled photon pair from two decohered photon pairs. Two polarization-entangled photon pairs are generated by spontaneous parametric down-conversion and then distributed through a channel that induces identical phase fluctuations to both pairs; this ensures that no entanglement is available as long as each pair is manipulated individually. Then, through collective local operations and classical communication we extract from the two decohered pairs a photon pair that is observed to be polarization-entangled. PMID- 12540895 TI - Direct observation of a local thermal vibration anomaly in a quasicrystal. AB - Quasicrystals have long-range order with symmetries that are incompatible with periodicity, and are often described with reference to a higher-dimensional analogue of a periodic lattice. Within the context of this 'hyperspace' crystallography, lattice dynamics of quasicrystals can be described by a combination of lattice vibrations and atomic fluctuations--phonons and phasons. However, it is difficult to see localized fluctuations in a real-space quasicrystal structure, and so the nature of phason-related fluctuations and their contribution to thermodynamic stability are still not fully understood. Here we use atomic-resolution annular dark-field scanning transmission electron microscopy to map directly the change in thermal diffuse scattering intensity distribution in the quasicrystal, through in situ high-temperature observation of decagonal Al72Ni20Co8. We find that, at 1,100 K, a local anomaly of atomic vibrations becomes significant at specific atomic sites in the structure. The distribution of these localized vibrations is not random but well-correlated, with a quasiperiodic length scale of 2 nm. We are able to explain this feature by an anomalous temperature (Debye-Waller) factor for the Al atoms that sit at the phason-related sites defined within the framework of hyperspace crystallography. The present results therefore provide a direct observation of local thermal vibration anomalies in a solid. PMID- 12540896 TI - Photocontrolled reversible release of guest molecules from coumarin-modified mesoporous silica. AB - Since the discovery of MCM-41 more than ten years ago, many investigations have explored the suitability of hexagonal mesoporous silicas for potential practical applications. These range from catalysis and optically active materials to polymerization science, separation technology and drug delivery, with recent successes in the fabrication of hybrid mesoporous organosilicas expected to open up further application possibilities. Because the pore voids of this class of materials exhibit relatively narrow pore size distributions in the range of 2-4 nm in diameter, mesoporous silicas can selectively include organic compounds and release them continuously at a later stage. The functionalization of MCM-41 pore voids with photoactive derivatives provides influence over the material's absorption behaviour, but full control over the release process remains difficult. Here we show that the uptake, storage and release of organic molecules in MCM-41 can be regulated through the photocontrolled and reversible intermolecular dimerization of coumarin derivatives attached to the pore outlets. Successful functionalization requires uncalcined MCM-41 still filled with the template molecules that directed the formation of its pores, to ensure that coumarin derivatives attach preferentially to the pore outlets, rather than their inside walls. We find that this feature and the one-dimensional, isolated nature of the individual pores allow for efficient and reversible photocontrol over guest access to the material's interior. PMID- 12540897 TI - Reductive dehalogenation of chlorinated dioxins by an anaerobic bacterium. AB - Polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs and PCDFs) are among the most notorious environmental pollutants. Some congeners, particularly those with lateral chlorine substitutions at positions 2, 3, 7 and 8, are extremely toxic and carcinogenic to humans. One particularly promising mechanism for the detoxification of PCDDs and PCDFs is microbial reductive dechlorination. So far only a limited number of phylogenetically diverse anaerobic bacteria have been found that couple the reductive dehalogenation of chlorinated compounds--the substitution of a chlorine for a hydrogen atom--to energy conservation and growth in a process called dehalorespiration. Microbial dechlorination of PCDDs occurs in sediments and anaerobic mixed cultures from sediments, but the responsible organisms have not yet been identified or isolated. Here we show the presence of a Dehalococcoides species in four dioxin-dechlorinating enrichment cultures from a freshwater sediment highly contaminated with PCDDs and PCDFs. We also show that the previously described chlorobenzene-dehalorespiring bacterium Dehalococcoides sp. strain CBDB1 (ref. 3) is able to reductively dechlorinate selected dioxin congeners. Reductive dechlorination of 1,2,3,7,8-pentachlorodibenzo-p-dioxin (PeCDD) demonstrates that environmentally significant dioxins are attacked by this bacterium. PMID- 12540898 TI - Directional postcopulatory sexual selection revealed by artificial insemination. AB - Postcopulatory sexual selection comprises both sperm competition, where the sperm from different males compete for fertilization, and cryptic female choice, where females bias sperm use in favour of particular males. Despite intense current interest in both processes as potential agents of directional sexual selection, few studies have attributed the success of attractive males to events that occur exclusively after insemination. This is because the interactions between pre- and post-insemination episodes of sexual selection can be important sources of variation in paternity. The use of artificial insemination overcomes this difficulty because it controls for variation in male fertilization success attributable to the female's perception of male quality, as well as effects due to mating order and the relative contribution of sperm from competing males. Here, we adopt this technique and show that in guppies, when equal numbers of sperm from two males compete for fertilization, relatively colourful individuals achieve greater parentage than their less ornamented counterparts. This finding indicates that precopulatory female mating preferences can be reinforced exclusively through postcopulatory processes occurring at a physiological level. Our analysis also revealed that relatively small individuals were advantaged in sperm competition, suggesting a possible trade-off between sperm competitive ability and body growth. PMID- 12540899 TI - Comparative power curves in bird flight. AB - The relationship between mechanical power output and forward velocity in bird flight is controversial, bearing on the comparative physiology and ecology of locomotion. Applied to flying birds, aerodynamic theory predicts that mechanical power should vary as a function of forward velocity in a U-shaped curve. The only empirical test of this theory, using the black-billed magpie (Pica pica), suggests that the mechanical power curve is relatively flat over intermediate velocities. Here, by integrating in vivo measurements of pectoralis force and length change with quasi-steady aerodynamic models developed using data on wing and body movement, we present mechanical power curves for cockatiels (Nymphicus hollandicus) and ringed turtle-doves (Streptopelia risoria). In contrast to the curve reported for magpies, the power curve for cockatiels is acutely concave, whereas that for doves is intermediate in shape and shows higher mass-specific power output at most speeds. We also find that wing-beat frequency and mechanical power output do not necessarily share minima in flying birds. Thus, aspects of morphology, wing kinematics and overall style of flight can greatly affect the magnitude and shape of a species' power curve. PMID- 12540900 TI - Neuronal synchrony does not correlate with motion coherence in cortical area MT. AB - Natural visual scenes are cluttered with multiple objects whose individual features must somehow be selectively linked (or 'bound') if perception is to coincide with reality. Recent neurophysiological evidence supports a 'binding-by synchrony' hypothesis: neurons excited by features of the same object fire synchronously, while neurons excited by features of different objects do not. Moving plaid patterns offer a straightforward means to test this idea. By appropriate manipulations of apparent transparency, the component gratings of a plaid pattern can be seen as parts of a single coherently moving surface or as two non-coherently moving surfaces. We examined directional tuning and synchrony of area-MT neurons in awake, fixating primates in response to perceptually coherent and non-coherent plaid patterns. Here we show that directional tuning correlated highly with perceptual coherence, which is consistent with an earlier study. Although we found stimulus-dependent synchrony, coherent plaids elicited significantly less synchrony than did non-coherent plaids. Our data therefore do not support the binding-by-synchrony hypothesis as applied to this class of motion stimuli in area MT. PMID- 12540901 TI - Selective gating of visual signals by microstimulation of frontal cortex. AB - Several decades of psychophysical and neurophysiological studies have established that visual signals are enhanced at the locus of attention. What remains a mystery is the mechanism that initiates biases in the strength of visual representations. Recent evidence argues that, during spatial attention, these biases reflect nascent saccadic eye movement commands. We examined the functional interaction of saccade preparation and visual coding by electrically stimulating sites within the frontal eye fields (FEF) and measuring its effect on the activity of neurons in extrastriate visual cortex. Here we show that visual responses in area V4 could be enhanced after brief stimulation of retinotopically corresponding sites within the FEF using currents below that needed to evoke saccades. The magnitude of the enhancement depended on the effectiveness of receptive field stimuli as well as on the presence of competing stimuli outside the receptive field. Stimulation of non-corresponding FEF representations could suppress V4 responses. The results suggest that the gain of visual signals is modified according to the strength of spatially corresponding eye movement commands. PMID- 12540902 TI - Pivotal role of oligomerization in expanded polyglutamine neurodegenerative disorders. AB - The expansion of a CAG repeat coding for polyglutamine in otherwise unrelated gene products is central to eight neurodegenerative disorders including Huntington's disease. It has been well documented that expanded polyglutamine fragments, cleaved from their respective full-length proteins, form microscopically visible aggregates in affected individuals and in transgenic mice. The contribution of polyglutamine oligomers to neurodegeneration, however, is controversial. The azo-dye Congo red binds preferentially to beta-sheets containing amyloid fibrils and can specifically inhibit oligomerization and disrupt preformed oligomers. Here we show that inhibition of polyglutamine oligomerization by Congo red prevents ATP depletion and caspase activation, preserves normal cellular protein synthesis and degradation functions, and promotes the clearance of expanded polyglutamine repeats in vivo and in vitro. Infusion of Congo red into a transgenic mouse model of Huntington's disease, well after the onset of symptoms, promotes the clearance of expanded repeats in vivo and exerts marked protective effects on survival, weight loss and motor function. We conclude that oligomerization is a crucial determinant in the biochemical properties of expanded polyglutamine that are central to their chronic cytotoxicity. PMID- 12540903 TI - A role for Drosophila LKB1 in anterior-posterior axis formation and epithelial polarity. AB - The PAR-4 and PAR-1 kinases are necessary for the formation of the anterior posterior (A-P) axis in Caenorhabditis elegans. PAR-1 is also required for A-P axis determination in Drosophila. Here we show that the Drosophila par-4 homologue, lkb1, is required for the early A-P polarity of the oocyte, and for the repolarization of the oocyte cytoskeleton that defines the embryonic A-P axis. LKB1 is phosphorylated by PAR-1 in vitro, and overexpression of LKB1 partially rescues the par-1 phenotype. These two kinases therefore function in a conserved pathway for axis formation in flies and worms. lkb1 mutant clones also disrupt apical-basal epithelial polarity, suggesting a general role in cell polarization. The human homologue, LKB1, is mutated in Peutz-Jeghers syndrome and is regulated by prenylation and by phosphorylation by protein kinase A. We show that protein kinase A phosphorylates Drosophila LKB1 on a conserved site that is important for its activity. Thus, Drosophila and human LKB1 may be functional homologues, suggesting that loss of cell polarity may contribute to tumour formation in individuals with Peutz-Jeghers syndrome. PMID- 12540904 TI - Activation of human CD4+ cells with CD3 and CD46 induces a T-regulatory cell 1 phenotype. AB - The immune system must distinguish not only between self and non-self, but also between innocuous and pathological foreign antigens to prevent unnecessary or self-destructive immune responses. Unresponsiveness to harmless antigens is established through central and peripheral processes. Whereas clonal deletion and anergy are mechanisms of peripheral tolerance, active suppression by T-regulatory 1 (Tr1) cells has emerged as an essential factor in the control of autoreactive cells. Tr1 cells are CD4+ T lymphocytes that are defined by their production of interleukin 10 (IL-10) and suppression of T-helper cells; however, the physiological conditions underlying Tr1 differentiation are unknown. Here we show that co-engagement of CD3 and the complement regulator CD46 in the presence of IL 2 induces a Tr1-specific cytokine phenotype in human CD4+ T cells. These CD3/CD46 stimulated IL-10-producing CD4+ cells proliferate strongly, suppress activation of bystander T cells and acquire a memory phenotype. Our findings identify an endogenous receptor-mediated event that drives Tr1 differentiation and suggest that the complement system has a previously unappreciated role in T-cell-mediated immunity and tolerance. PMID- 12540907 TI - Quiet debut for the double helix. AB - Past discoveries usually become aggrandized in retrospect, especially at jubilee celebrations, and the double helix is no exception. The historical record reveals a muted response by the scientific community to the proposal of this structure in 1953. Indeed, it was only when the outlines appeared of a mechanism for DNA's involvement in protein synthesis that the biochemical community began to take a serious interest in the structure. PMID- 12540908 TI - Discovering genes are made of DNA. AB - Maclyn McCarty is the sole surviving member of the team that made the remarkable discovery that DNA is the material of inheritance. This preceded by a decade the discovery of the structure of DNA itself. Here he shares his personal perspective of those times and the impact of the double helix. PMID- 12540909 TI - The double helix and the 'wronged heroine'. AB - In 1962, James Watson, Francis Crick and Maurice Wilkins received the Nobel prize for the discovery of the structure of DNA. Notably absent from the podium was Rosalind Franklin, whose X-ray photographs of DNA contributed directly to the discovery of the double helix. Franklin's premature death, combined with misogynist treatment by the male scientific establishment, cast her as a feminist icon. This myth overshadowed her intellectual strength and independence both as a scientist and as an individual. PMID- 12540910 TI - The mosaic that is our genome. AB - The discovery of the basis of genetic variation has opened inroads to understanding our history as a species. It has revealed the remarkable genetic similarity we share with other individuals as well as with our closest primate relatives. To understand what make us unique, both as individuals and as a species, we need to consider the genome as a mosaic of discrete segments, each with its own unique history and relatedness to different contemporary and ancestral individuals. PMID- 12540911 TI - Nature, nurture and human disease. AB - What has been learnt about individual human biology and common diseases 50 years on from the discovery of the structure of DNA? Unfortunately the double helix has not, so far, revealed as much as one would have hoped. The primary reason is an inability to determine how nurture fits into the DNA paradigm. We argue here that the environment exerts its influence at the DNA level and so will need to be understood before the underlying causal factors of common human diseases can be fully recognized. PMID- 12540912 TI - The double helix in clinical practice. AB - The discovery of the double helix half a century ago has so far been slow to affect medical practice, but significant transformations are likely over the next 50 years. Changes to the way medicine is practised and new doctors are trained will be required before potential benefits are realized. PMID- 12540913 TI - The Mona Lisa of modern science. AB - No molecule in the history of science has reached the iconic status of the double helix of DNA. Its image has been imprinted on all aspects of society, from science, art, music, cinema, architecture and advertising. This review of the Mona Lisa of science examines the evolution of its form at the hands of both science and art. PMID- 12540914 TI - Portrait of a molecule. AB - The double helix is idealized for its aesthetic elegant structure, but the reality of DNA's physical existence is quite different. Most DNA in the cell is compressed into a tangled package that somehow still exposes itself to meticulous gene-regulatory control. Philip Ball holds a mirror up to what we truly know about the mysteries of DNA's life inside a cell. PMID- 12540915 TI - Ten years of tension: single-molecule DNA mechanics. AB - The basic features of DNA were elucidated during the half-century following the discovery of the double helix. But it is only during the past decade that researchers have been able to manipulate single molecules of DNA to make direct measurements of its mechanical properties. These studies have illuminated the nature of interactions between DNA and proteins, the constraints within which the cellular machinery operates, and the forces created by DNA-dependent motors. PMID- 12540916 TI - DNA in a material world. AB - The specific bonding of DNA base pairs provides the chemical foundation for genetics. This powerful molecular recognition system can be used in nanotechnology to direct the assembly of highly structured materials with specific nanoscale features, as well as in DNA computation to process complex information. The exploitation of DNA for material purposes presents a new chapter in the history of the molecule. PMID- 12540917 TI - DNA replication and recombination. AB - Knowledge of the structure of DNA enabled scientists to undertake the difficult task of deciphering the detailed molecular mechanisms of two dynamic processes that are central to life: the copying of the genetic information by DNA replication, and its reassortment and repair by DNA recombination. Despite dramatic advances towards this goal over the past five decades, many challenges remain for the next generation of molecular biologists. PMID- 12540918 TI - DNA damage and repair. AB - The aesthetic appeal of the DNA double helix initially hindered notions of DNA mutation and repair, which would necessarily interfere with its pristine state. But it has since been recognized that DNA is subject to continuous damage and the cell has an arsenal of ways of responding to such injury. Although mutations or deficiencies in repair can have catastrophic consequences, causing a range of human diseases, mutations are nonetheless fundamental to life and evolution. PMID- 12540919 TI - The double helix and immunology. AB - The immune system can recognize and produce antibodies to virtually any molecule in the Universe. This enormous diversity arises from the ingenious reshuffling of DNA sequences encoding components of the immune system. Immunology is an example of a field completely transformed during the past 50 years by the discovery of the structure of DNA and the emergence of DNA technologies that followed. PMID- 12540920 TI - The digital code of DNA. AB - The discovery of the structure of DNA transformed biology profoundly, catalysing the sequencing of the human genome and engendering a new view of biology as an information science. Two features of DNA structure account for much of its remarkable impact on science: its digital nature and its complementarity, whereby one strand of the helix binds perfectly with its partner. DNA has two types of digital information--the genes that encode proteins, which are the molecular machines of life, and the gene regulatory networks that specify the behaviour of the genes. PMID- 12540921 TI - Controlling the double helix. AB - Chromatin is the complex of DNA and proteins in which the genetic material is packaged inside the cells of organisms with nuclei. Chromatin structure is dynamic and exerts profound control over gene expression and other fundamental cellular processes. Changes in its structure can be inherited by the next generation, independent of the DNA sequence itself. PMID- 12540922 TI - Alternative approaches. PMID- 12540923 TI - The birth of biotechnology. PMID- 12540924 TI - Abrupt changes in the Asian southwest monsoon during the Holocene and their links to the North Atlantic Ocean. AB - During the last ice age, the Indian Ocean southwest monsoon exhibited abrupt changes that were closely correlated with millennial-scale climate events in the North Atlantic region, suggesting a mechanistic link. In the Holocene epoch, which had a more stable climate, the amplitude of abrupt changes in North Atlantic climate was much smaller, and it has been unclear whether these changes are related to monsoon variability. Here we present a continuous record of centennial-scale monsoon variability throughout the Holocene from rapidly accumulating and minimally bioturbated sediments in the anoxic Arabian Sea. Our monsoon proxy record reveals several intervals of weak summer monsoon that coincide with cold periods documented in the North Atlantic region--including the most recent climate changes from the Medieval Warm Period to the Little Ice Age and then to the present. We therefore suggest that the link between North Atlantic climate and the Asian monsoon is a persistent aspect of global climate. PMID- 12540926 TI - Individual rooms in the NICU - an evolving concept. AB - The provision of individual rooms for NICU patients has several attractive benefits, including the ability to provide the appropriate environmental stimuli for each child, and increased privacy and accessibility for families. This concept can put serious strain on caregivers, however, by isolating them from one another and from the multiple infants for whom they may have responsibility, and places additional financial and space demands on the institution. These problems are not insurmountable, however, and use of individual rooms for at least some patients is feasible and probably desirable if certain considerations are addressed in the planning and design process. PMID- 12540928 TI - The President's New Year message. PMID- 12540929 TI - In-flight oral-facial pain. PMID- 12540930 TI - New horizons in the regulation of dentistry. AB - I'm delighted to have the opportunity to share with you my understanding of the much-needed and long-awaited reform of the General Dental Council. Change is moving apace and the new, modern GDC, for which the existing Council has been working so hard, is taking shape as part of widespread fundamental reforms in the regulation of healthcare. PMID- 12540932 TI - Further statistics in dentistry. Part 7: repeated measures. AB - Consider the situation in which there is a single quantitative variable of interest that is measured on each individual on several different occasions. Typically, these occasions are defined time points (usually including pre treatment as well as various post-treatment times), so that each individual contributes a series of readings. The main objective of the study may be to compare the responses on this variable when each individual has been assigned to one of two or more treatments groups. PMID- 12540933 TI - A suspected case of tuberculosis of the temporomandibular joint. AB - Primary tuberculosis is symptomless in the great majority of individuals. The clinical appearance of tuberculosis (TB) of the temporomandibular joint is similar to that of ordinary arthritis of this joint and so, is unlikely to be diagnosed early in non-destructive disease, especially when there are no symptoms and signs of TB elsewhere in the body. The role of Mycobacterium tuberculosis and other opportunistic organisms should not be overlooked in complicated head and neck infections as this is an ever increasing problem today, especially due to the emergence of multi-drug resistant strains of tuberculosis in some urban areas. PMID- 12540934 TI - A view from the bodies corporate. 5. Integrated dental holdings plc. Interview by F. Stuart-Wilson. AB - My interview with David Hudaly, Chief Executive of IDH plc took place at their unassuming head office in Bolton. I make the mistake of assuming, like many, that IDH's operations are based mainly in the North. David Hudaly puts me right fairly early on in our interview. In fact IDH is not one, but two bodies corporate now with 146 practices across the UK, and incorporating the Whitecross and Petrie Tucker chains (although some of the Whitecross units have recently been sold). The business structure appears at first sight to be more complex than the other corporates I have encountered in this series; IDH floated in February 2002 and is now a plc. PMID- 12540938 TI - General dental practitioners' knowledge of and attitudes towards the employment of dental therapists in general practice. AB - OBJECTIVE: To investigate general dental practitioners' knowledge of and attitudes towards dental therapists, to ascertain the likelihood of their employment in general dental practice, what client groups they would be likely to treat, and to identify the main perceived barriers to their employment in general dental practice. METHOD: Postal questionnaire. SETTING: General dental practitioners in the county of West Sussex. SAMPLING: All dentists holding a contract to provide general dental services in West Sussex were contacted. Final sample size was 200. KEY FINDINGS: Thirty eight per cent of dentists said they would employ a therapist if legislation allowed. Main perceived barriers were cost, lack of knowledge and dentists' acceptance. CONCLUSIONS: In general dentists had a favourable attitude towards dental therapists, although there was a real lack of knowledge about their permitted duties. Most dentists felt therapists should treat children and people with special needs. PMID- 12540939 TI - Patient-maintained sedation for oral surgery using a target-controlled infusion of propofol - a pilot study. AB - OBJECTIVE: To assess the safety and efficacy of a new patient-maintained propofol system for conscious sedation in dentistry. DESIGN: Prospective clinical trial SETTING: Department of Sedation, Glasgow Dental Hospital and School, 2001 SUBJECTS AND METHODS: Patients scheduled for oral surgery with conscious sedation. Exclusions included ASA IV -V, inability to use the handset, opioid use and severe respiratory disease. INTERVENTIONS: Patients were given intravenous propofol to a level of 1.0 microg/ml (reducing from 1.5 microg/ml) using a target controlled infusion system, they then controlled their sedation level by double clicking a handset which on each activation increased the propofol concentration by 0.2 microg/ml. MAIN OUTCOME MEASURES: Oxygen saturation, patient satisfaction, and surgeon satisfaction. RESULTS: Twenty patients were recruited, 16 female and four male. Nineteen patients completed sedation and treatment successfully. Mean lowest oxygen saturation was 94%. No patients were over-sedated. All patients successfully used the system to maintain a level of sedation adequate for their comfort. Patient and surgeon satisfaction were consistently high. CONCLUSIONS: Initial experience with this novel system has confirmed safety, patient satisfaction and surgeon satisfaction. PMID- 12540940 TI - Participation of UK dentists in continuing professional development. AB - INTRODUCTION: This paper reports participation of dentists in continuing professional development (CPD) and factors affecting participation. METHOD: All general dental practitioners (GDPs) in three deaneries in England were surveyed. The overall response rate was 54% (n = 2082); by deanery it was 68% West Midlands, 45% South West and 44% Anglia. Findings across deaneries were remarkably similar. Comparisons with national data show no notable bias in the sample for gender, owners/partners and age/experience. RESULTS: Most frequent forms of CPD were journal reading and courses in which almost all engaged. A score based on individual participation in CPD over the 12-month period was calculated. The mean score (hours) for participation in verifiable CPD was 31 (median 25) and for general, 29 (median 29). In terms of the GDC's Lifelong Learning Scheme, 57% were already undertaking 50 hours. Net of other effects, those less likely to be doing 50 hours are those with more years in practice and single-handed practitioners. Greater access to courses and media-based CPD is desired. CONCLUSION: Certain groups of dentists will need support to meet the requirements of the GDC's Lifelong Learning Scheme. Statutory peer review or clinical audit will significantly alter the CPD profile of most dentists. This has implications for facilitators. PMID- 12540943 TI - Well! Am I afraid of dentists? AB - Do you think about what it's like at the other end of the drill? A patient tells it how it is from her point of view. PMID- 12540946 TI - Intravascular tissue factor pathway--a model for rapid initiation of coagulation within the blood vessel. AB - The loss of blood through vessel wall ruptures is initially prevented by the rapid adhesion of platelets to the subendothelium, and the formation of a thrombus consisting of platelets and different types of leukocytes. Concomitantly, the coagulation process is thought to be activated by vascular wall tissue factor (TF). Here, a new model for the initiation of coagulation is presented, based on unexpected findings on the presence and functional activation of TF within the blood itself. TF was recently found to be stored in the alpha granules of resting platelets under physiological conditions. Activation by collagen exposes TF on the platelet cell membrane and on platelet derived microvesicles. Adhesive interactions of the TF bearing platelets and microvesicles to neutrophils and monocytes support the functional activation of the blood based TF. The intravascular TF pathway is proposed to play a significant role during hemostasis by enabling the generation of fibrin at the site of the developing thrombus. PMID- 12540947 TI - Antithrombin I. Inhibition of thrombin generation in plasma by fibrin formation. AB - Thrombin substrate binding is mediated through fibrinogen recognition "exosite 1" in thrombin, resulting in fibrinopeptide cleavage to form fibrin. In addition, thrombin exhibits "non-substrate" binding to fibrin, an activity termed "Antithrombin I". Antithrombin I (AT-I) is characterized by two classes of thrombin binding sites, the first of "low affinity" in the fibrin E domain, and the other of high affinity, that is situated between C-terminal residues 414 and 427 of a variant gamma chain termed gamma'(1-427L), Plasma fibrinogen molecules containing gamma' chains ("fibrinogen 2") are virtually all heterodimers containing one gamma(A) chain (platelet-binding) and one gamma' chain. The remaining fibrinogen (approximately 85%) is homodimeric, lacks high affinity thrombin-binding potential, and is termed " fibrinogen 1" (gamma(A)/gamma(A)). Thrombin generation in recalcified fibrinogen-depleted or congenital afibrinogenemic plasma is increased. Repletion with fibrinogen 1 has a modest effect in normalizing thrombin generation, whereas repletion with fibrinogen 2 (gamma(A)/gamma') has a more marked effect. A post-translational gamma' chain derivative, gamma'(1-423P), accounts for 3%-34% of the gamma' chain population, lacks thrombin binding potential, and arises by proteolytic processing at the expense of gamma' (1-427L) chains. Little is known about its effect on plasma AT I activity under normal or pathological circumstances. In summary, fibrin formation (Antithrombin I) inhibits thrombin generation in clotting blood by sequestering thrombin, and "high-affinity" thrombin-binding (i.e., via gamma' chains) plays a dominant role in this process. AT-1 should be considered when assessing the pathogenesis of thromboembolic disease. PMID- 12540948 TI - Leukocyte trans-endothelial migration: JAMs add new pieces to the puzzle. AB - The molecular details of leukocyte transmigration through the endothelial barrier (also called diapedesis), which is the final step of leukocyte extravasation from the circulation to a given site of inflammation, are by far not well understood. The present review will focus on the different mechanisms potentially involved in leukocyte trans-endothelial migration. Both homophilic and heterophilic interactions between leukocyte and endothelial cell receptors will be covered, with a particular focus on the growing gene family of junctional adhesion molecules (JAM). Deciphering their mechanisms of interaction will also allow to unravel novel strategies for therapeutic intervention in inflammatory or atherothrombotic diseases. PMID- 12540949 TI - Interaction of pathogens with the endothelium. AB - The endothelium lines the inner surface of the vessel wall establishing a multifunctional, semi-permeable cellular barrier at blood-tissue interface. The large total surface of the endothelium is exposed to pathogens, pathogen-derived products as well as to agents of the activated host defense during an inflammatory reaction. The endothelium is not only specifically targeted by important infective agents like Rickettsiae (1) or Bartonella (2), it is involved in virtually most, if not all, acute inflammatory responses. Pathogens attack the endothelium by a wide variety of strategies, as different as activation of preformed receptor-mediated pathways in the endothelium, release of pore-forming exotoxins or intracellular replication and chronic parasitism. These pathophysiological forces affect the endothelial phenotype, resulting in endothelial barrier dysfunction, increased leukocyte-endothelial interaction, mediator release, and procoagulant activity. Moreover, endothelial responses retroact on the invading pathogen as well as on the host defense resulting in a complex and dynamic interaction. Endothelial activation contributes considerably to inflammation and resulting clinical characteristics. In this context the endothelium is not just a passive victim, it rather aggravates the ongoing struggle with the pathogen. In this review we focus on some important mechanisms of the cellular microbiology of endothelial infection by bacteria and viruses. PMID- 12540950 TI - Magnetic resonance imaging of tumor vasculature. AB - Angiogenic activity and formation of a vascular network facilitate tumor perfusion and play a critical role in tumor growth and metastasis. Tumor vasculature may be visualized by means of parametric imaging of specific morphological and physiological characteristics that collectively describe its properties. In this review, we describe advanced magnetic resonance imaging (MRI) techniques that have been developed in order to image and quantify the distribution of tumor vasculature throughout the tumor and characterize its function. These techniques have been used to monitor changes in the magnetic resonance signal intensity of tissue water hydrogens generated by intrinsic effects, as well as by exogenous contrast agents administered into the blood circulation. We further describe specific applications of magnetic resonance imaging using a contrast agent, gadolinium diethylene triamine penta-acetic acid (GdDTPA), which has long been approved for clinical use. Examples include studies of the vascular properties of breast cancer tumors and metastases in animal models, as well as of breast cancer vasculature in patients. We also discuss the use of MRI to improve breast cancer diagnosis in humans by quantifying the permeability of the tumor vasculature. By maximizing the spatial resolution of the images in both animal and human studies, the capacity of magnetic resonance imaging to enhance our understanding of the processes regulating tumor angiogenesis, and improve the diagnosis of cancer, could be clearly demonstrated. PMID- 12540951 TI - Orthopaedic surgery in persons with haemophilia. AB - The orthopaedic problems of haemophilia and the surgical techniques most frequently performed by orthopaedic surgeons on haemophilia patients are summarised in this review article. Sometimes it can be recommendable to carry out two or three orthopaedic surgical procedures in a single operative session, with the aim of solving the functional problem in a more global way, which usually is polyarticular. There is no doubt that such an undertaking implies a greater anaesthetic risk, but also avoids the repetition of surgical procedures and saves factor consumption. Close cooperation between haematologists, orthopaedic surgeons, rehabilitation physicians, paediatricians, psychologists, physiotherapists and nurses is paramount for the satisfactory result of any orthopaedic surgical procedure. There is no doubt that continuous prophylactic clotting factor replacement (prophylactic therapy) is the optimal way to avoid the orthopaedic problems of haemophilia that can still be seen today. Until such a goal can be reached, the orthopaedic surgeon will continue performing arthrocentheses, synoviortheses, synovectomies, tendon lengthenings, debridements, osteotomies, joint replacements, osteosynthesis of fractures and other less frequent surgical procedures on persons with haemophilia. PMID- 12540952 TI - The influence of exogenous magnesium chloride on the apparent INR determined with human, rabbit, and bovine thromboplastin reagents. AB - Magnesium ions can shorten the tissue factor-induced coagulation time. Some blood collection systems with sodium citrate are contaminated with variable amounts of magnesium and influence the results of the prothrombin time (PT) test. The aim of the study was to determine the dose-response relationship between exogenous magnesium chloride added to blood and the PT and the international normalized ratio (INR). Blood specimens from twenty patients on oral anticoagulant therapy were investigated. Four different types of thromboplastin reagents were used: recombinant human, human placenta, rabbit brain, and bovine brain combined with adsorbed bovine plasma. With all four reagents, exogenous magnesium induced a reduction of the apparent INR. Bovine thromboplastin was not as responsive to magnesium as the human and rabbit reagents. The magnitude of the INR deviation induced by 0.1 mmol/l magnesium in the blood was smaller than 10% in all patient samples. At 0.5 mmol/l magnesium in the blood, 10-35% of the patient samples had INR deviations greater than 10%, depending on the thromboplastin reagent used. PMID- 12540953 TI - The circulatory half-lives of alpha-profibrin and alpha-fibrin monomer, and comparisons with other fibrin(ogen) derivatives. AB - Previous studies showed that alpha-fibrin monomer (lacking both A fibrinopeptides, FPA) is normally cleared from the circulation before it assembles into a clot. Recent studies indicate that substantial quantities of an intermediate, alpha-profibrin lacking only one of the two FPA are produced in the course of conversion of human fibrinogen to fibrin. Since clearance of the alpha fibrin monomer is saturable and receptor mediated, the extent to which alpha profibrin or other fibrin(ogen) derivatives might compete for monomer uptake was deemed important. We compared plasma decay of injected human alpha-fibrin, fibrinogen, and alpha-profibrin in rabbits using rabbit anti-human fibrinogen for assays. The circulatory half-life of human alpha-fibrin monomer was short (t(1/2) = 2.3 h) and followed a simple exponential decay curve, as anticipated from clearance of rabbit alpha-fibrin. It was absorbed as fast as it permeated the extravascular space with no redistribution. Human fibrinogen had a long half-life (t(1/2) = 39.5 h), calculated from the double exponential plasma decay curves (redistribution + catabolism) observed over 28 h. The alpha-profibrin had an intermediary half-life (t(1/2) = 11 h) determined from double exponential decay curves. Since redistribution accompanied the slow clearance of alpha-profibrin, its binding by the fibrin receptor(s) must be weak, probably too weak to compete with the clearance of alpha-fibrin monomer. The initial production of alpha fibrin monomer is only partially dependent on prior formation of alpha-profibrin, as recently shown. Thus, it is the slow clearance and the weak competition from alpha-profibrin that underlie the occurrence of substantial levels of alpha profibrin unaccompanied by detectable levels of alpha-fibrin monomer in many subjects with vascular disease. PMID- 12540954 TI - Chemical crosslinking of urokinase to pulmonary surfactant protein B for targeting alveolar fibrin. AB - Intraalveolar fibrin formation is a consistent finding in acute inflammatory and chronic interstitial lung disease. Polymerization of fibrin in the presence of pulmonary surfactant results in far-reaching incorporation of the hydrophobic surfactant compounds into the growing fibrin matrix, with loss of surface activity, altered fibrin structure and reduced susceptibility of the clot to fibrinolysis. For specific targeting of such alveolar fibrin, we designed a hybrid molecule consisting of the catalytic domain of urokinase (B-chain) and the hydrophobic surfactant protein B (SP-B), termed SPUC. The urokinase B-chain, obtained by limited reduction of human two-chain-urokinase (u-PA) and subsequent affinity purification, was chemically coupled to SP-B in a semi-organic solvent system using a hetero-bifunctional crosslinker. Purification of the chimeric proteins included reversed phase and cation exchange chromatography. SDS-PAGE and Western Blotting with immunostaining were employed for biochemical characterization of the conjugate. Chromogenic substrate assays, (125)I-based fibrin plate assays, active site titration and surface tension measurements (pulsating bubble surfactometer) were performed to analyze the specific fibrinolytic activity of the conjugate and its surface activity. SPUC was found i) to be assembled stoichiometrically in a 1: 1 fashion (SP-B: u-PA), ii) to fully retain the biophysical activity as compared to native SP-B and iii) to also retain the fibrinolytic activity. SPUC was 2-3 fold more effective in lysis of surfactant containing clots and 5-fold more resistant against plasminogen activator 1 (PAI-1) as compared to the native u-PA. We conclude that urokinase and SP-B can be chemically crosslinked, thereby yielding a fibrinolytic enzyme suitable for targeting alveolar fibrin. PMID- 12540955 TI - Localization of tissue factor pathway inhibitor to lipid rafts is not required for inhibition of factor VIIa/tissue factor activity. AB - Tissue factor pathway inhibitor (TFPI) abrogates coagulation initiated by the factor VIIa/tissue factor catalytic complex. While the gene structure of TFPI suggests that it is a secreted protein, a large pool of TFPI is associated with the vascular endothelium through its affinity for a glycosylphosphati-dylinositol (GPI)-linked membrane protein. Inhibition of tissue factor by TFPI coincides with the translocation of quaternary complexes containing tissue factor, factor VIIa, factor Xa, and TFPI to detergent-insoluble plasma membrane domains rich in cholesterol, sphingomyelin, and GPI-linked proteins known as lipid rafts and caveolae. It is not known if localization of TFPI to these membrane domains is required for its inhibition of tissue factor procoagulant activity. We generated chimeric TFPI molecules linked directly to the plasma membrane via a GPI anchor or hydrophobic transmembrane domain and expressed these in HEK293 cells that produce tissue factor but not endogenous TFPI. The GPI-anchored chimera was exclusively enriched in detergent-insoluble membrane fractions while the transmembrane molecule was not. Transfectants expressing equal levels of the GPI linked or transmembrane TFPI displayed equal anticoagulant potency as assessed by tissue factor-mediated conversion of factor X to factor Xa. Disruption of lipid rafts with cyclodextrin likewise had no effect on the inhibitory activity of the transmembrane or GPI-linked TFPI chimeras in HEK293 cells, nor on endogenous TFPI expressed by ECV304 cells. Thus, we conclude that the GPI anchor and membrane localization to lipid rafts does not enhance inhibition of factor VIIa/tissue factor by cell-surface associated TFPI. PMID- 12540956 TI - Elucidation of the paratope of scFv-8H9D4, a PAI-1 neutralizing antibody derivative. AB - Interfering with increased levels of plasminogen activator inhibitor-1 (PAI-1) might offer new therapeutic strategies for a variety of cardiovascular diseases. Inactivation of PAI-1 can be accomplished by a number of monoclonal antibodies (MA), including MA-8H9D4. In a previous study, a single-chain variable fragment (scFv-8H9D4) was cloned and found to have the same properties as the parental MA 8H9D4. In the present study, we identified the residues of scFv-8H9D4 that contribute significantly to the paratope. The complementarity determining region 3 from the heavy (H3) and the light (L3) chain were analysed through site directed mutagenesis. Out of twelve mutations, only four residues appeared to contribute to the paratope. The affinity of scFv-8H9D4-H3-L97D for PAI-1 was 38 fold decreased (K(A) = 4.8 +/- 0.2 x 10(7) M(-1) vs. 1.8 +/- 0.7 x 10(9) M(-1) for scFv-8H9D4) whereas scFv-8H9D4-H3-R98Y did not bind to PAI-1. The affinities of scFv-8H9D4-L3-Y91S and scFv-8H9D4-L3-F94D for PAI-1 were 9- and 5-fold reduced, respectively, whereas the combined mutation resulted in an 86-fold decreased affinity (K(A) = 2.1 +/- 0.2 x 10(7) M(-1)). In accordance with the affinity data, these mutants had no, or a reduced, PAI-1 inhibitory capacity, confirming that these four particular residues form the major interaction site of scFv-8H9D4 with PAI-1. In combination with the three-dimensional structure, these data contribute to the rational design of PAI-1 neutralizing compounds. PMID- 12540957 TI - Prolonged elevations in haemostatic and rheological responses following psychological stress in low socioeconomic status men and women. AB - Low socioeconomic status (SES) and psychological stress are associated with increased risk of coronary heart disease, and both may influence haemostatic responses. Von Willebrand factor (vWF), Factor VIII, plasma viscosity, haematocrit, blood viscosity, tissue plasminogen activator (t-PA) and fibrin D dimer were measured at rest and following stressful tasks in 238 middle-aged British civil servants. SES was defined by grade of employment. Lower SES was associated with higher resting vWF, Factor VIII and plasma viscosity. Psychological stress stimulated increases in haemostatic and rheological factors. Initial stress responses did not vary with SES, but Factor VIII, plasma viscosity and blood viscosity remained more elevated 45 minutes post-stress in lower SES participants. High blood pressure stress reactivity was also associated with greater haemostatic responses. We conclude that lower SES is characterised by more prolonged elevations in procoagulant responses following psychological stress, and that these processes might contribute to increased cardiac risk. PMID- 12540958 TI - Peripheral artery disease in type 2 diabetes: the role of fibrinolysis. AB - The aim of the present study is to verify the relationship between peripheral artery disease (PAD) and some coagulation/fibrinolysis parameters in type 2 diabetic patients. Sixty-three type 2 diabetic patients, without PAD, were studied at baseline and after 4 years. Assessments included tissue-Plasminogen Activator (t-PA), Plasminogen Activator Inhibitor-1 antigen (PAI-1 Ag), Plasminogen Activator Inhibitor-1 activity (PAI-1 Act), Plasminogen (Pl), Fibrin peptide A (FPA), Fibrinogen (Fr), and the ankle/brachial pressure index (ABI). We observed a significant difference between diabetic patients and controls as regards tPA (11.8 +/- 5.4 vs. 6.6 +/- 3.0 ng/ml; p <0.05) and PAI-1 Act (17.8 +/- 9.2 vs. 11.7 +/- 6.6 ng/dl; p <0.005). After 4 years 13 diabetic patients became vasculopathic and, at baseline, had significantly lower tPA (8.9 +/- 4.8 vs. 12.5 +/- 5.3; p <0.011), and higher PAI-1 Ag (50.8 +/- 22.2 vs. 32 +/- 22.2; p <0.006), and PAI-1 Act values (24.1 +/- 9.5 vs. 16.1 +/- 8.4; p <0.014), compared with 50 diabetic patients who did not develop PAD after 4 years. These data show that the physiological equilibrium which exists between t-PA and PAI-1 moves towards higher levels in our diabetic patients compared with controls, at baseline, whereas diabetic patients who developed PAD showed a shift towards an antifibrinolytic pathway with diminished t-PA, increased PAI-1 Ag and PAI-1 Act and consequently procoagulant activity. Our study suggests that hypofibrinolysis may be involved in the future onset of PAD in type 2 diabetic patients. PMID- 12540959 TI - Ruling out clinically suspected pulmonary embolism by assessment of clinical probability and D-dimer levels: a management study. AB - D-dimer test combined with clinical probability assessment has been proposed as the first step in the diagnostic work-up of patients with suspected pulmonary embolism (PE). In a prospective management study we investigated the safety and efficiency of excluding PE by a normal D-dimer combined with a low or moderate clinical probability. Of the 202 study patients this combination ruled out PE in 64 (32%) patients. The 3-month thromboembolic risk in these patients was 0% (95% CI, 0.0-5.6%). The prevalence of PE in the entire cohort was 29% (59 patients), whereas in the low, moderate and high clinical probability groups this was 25%, 26% and 50%, respectively. We conclude that ruling out suspected PE by a normal D dimer combined with a low or moderate clinical probability appears to be a safe and efficient strategy. The accuracy of the clinical probability assessment is modest. PMID- 12540960 TI - Antithrombotic activity of LB30057, a newly synthesized direct thrombin inhibitor. AB - An amidrazonophenylalanine derivative, LB30057, inhibits the catalytic activity of thrombin potently by interaction with the active site of thrombin, and has high water solubility. In the present study, we evaluated the effect of LB30057 on the biological activities of thrombin at various tissues, and determined whether thrombin inhibition by LB30057 could reduce the incidence of occlusive thrombosis in an in vivo animal model. Treatment with LB30057 to human plasma prolonged clotting times in a concentration-dependent manner. LB30057 suppressed significantly thrombin-induced phosphatidylserine (PS) exposure in platelets, suggesting that LB30057 could inhibit blood coagulation accelerated by PS exposure. In human platelets, soluble thrombin- and clot-induced platelet aggregation was inhibited by LB30057 potently. Consistent with this finding, LB30057 showed concentration-dependent inhibitory effects on serotonin secretion and P-selectin expression induced by thrombin in platelets. In the blood vessel isolated from the guinea pig, treatment with LB30057 resulted in a concentration dependent inhibition of thrombin-induced vascular contraction. In vivo study revealed that LB30057 following oral administration significantly increased the time to occlusion and improved carotid arterial patency using rat carotid artery thrombosis model. All these results suggest that LB30057 is a potent inhibitor of biological activities of thrombin at various target tissues and, therefore, might be developed as an antithrombotic agent for treatment and prevention of thrombotic diseases. PMID- 12540961 TI - DX-9065a inhibition of factor Xa and the prothrombinase complex: mechanism of inhibition and comparison with therapeutic heparins. AB - Factor Xa (fXa) is the key enzyme of the prothrombinase complex that generates thrombin hence it is a good target for antithrombotic therapy. Here, the anti-fXa and anti-prothrombinase activities of DX-9065a which is an active-site directed inhibitor of fXa, and therapeutic heparins which are dependent on antithrombin (AT) for their anticoagulant function, were studied in amidolytic and proteolytic activity assays. It was found that DX-9065a is a competitive inhibitor of the Spectrozyme FXa (SpFXa) cleavage by both fXa and prothrombinase with similar K(i) values of approximately 10-20 nM. However, DX-9065a acted as a non-competitive inhibitor of prothrombin activation by prothrombinase with a Ki of approximately 26 nM. On the other hand, therapeutic heparins were effective catalysts of both fXa and prothrombinase inhibition by AT in the presence of SpFXa, but were ineffective in the presence of prothrombin. Further studies revealed that Tyr(99), a residue in the extended S2-S4 binding pocket of fXa, plays a key role in determination of specificity of the DX-9065a interaction. PMID- 12540962 TI - Plasma membrane Ca2+-ATPase isoform 4b is phosphorylated on tyrosine 1176 in activated human platelets. AB - Plasma membrane Ca(2+) -ATPase isoform 4b (PMCA4b) is phosphorylated on a tyrosine residue during platelet activation resulting in inhibition of its ATPase activity. We now report that tyrosine 1176 (Y(1176)) in the carboxyl (C-) terminal domain of PMCA4b is the phosphorylated residue. Two tyrosine residues located in the C-terminus of PMCA4b, Y(1122) and Y(1176) can be removed by calpain-dependent cleavage. This truncation removes all of the tyrosine phosphates added to PMCA during platelet activation. Sequence analysis indicates that Y(1176) is a likely substrate for focal adhesion kinase (FAK), while Y(1122) is not located in a tyrosine phosphorylation motif. This is the same residue we reported earlier to be phosphorylated by Src kinase in vitro. Thus we conclude that Y(1176) is the only tyrosine phosphorylated during platelet activation. Results of co-immunoprecipitation, treatment with tyrosine kinase inhibitors and integrin inhibition experiments suggest that FAK is responsible for PMCA4b tyrosine phosphorylation during platelet activation. PMID- 12540963 TI - Platelet phosphatidylserine exposure and procoagulant activity in clotting whole blood--different effects of collagen, TRAP and calcium ionophore A23187. AB - We have studied the effects of different platelet agonists on phosphatidylserine (PS) exposure and clotting times in blood without anticoagulants. Similar reductions in clotting time were obtained for collagen, TRAP-6 or calcium ionophore A23187 (50 micro mol/L), in spite of huge differences in PS expression [6.7 +/- 2.4%, 2.3 +/- 0.5% and 99.9 +/- 0.1%, respectively (mean +/- SD, n = 5)]. Furthermore, the clotting times were much longer for samples with A23187 exposing the same amounts of PS as samples with collagen or TRAP-6. Annexin V reversed the clotting time reduction, but could not prevent coagulation. Addition of phospholipid vesicles containing 20% PS neither affected the clotting times nor induced clotting in recalcified, platelet-free plasma. We conclude that platelet PS exposure is necessary, but not sufficient, for the coagulation amplification observed when platelets are stimulated via physiological receptors in a whole blood environment. PMID- 12540964 TI - Diabetes duration may modify the association between genetic variation in the glycoprotein Ia subunit of the platelet collagen receptor and risk of severe diabetic retinopathy: a working hypothesis. AB - Genetic factors appear to contribute to the severity and progression of diabetic retinopathy. We assessed the associations of the C807T and Glu505Lys variants of the glycoprotein Ia (alpha(2) integrin) subunit of the platelet/endothelial collagen receptor and risk of retinopathy in a population-based survey of 288 diabetic patients in one Swedish community. Neither variant was associated with retinopathy risk overall. However, the 807T variant was associated with increased risk of severe retinopathy, and the association was modified by diabetes duration. Among patients with diabetes of longer duration (>/=25 years), the 807T variant was strongly associated with risk of severe retinopathy (odds ratio 7.49, 95% confidence interval 1.75 to 32.1). There was no association between the 807T variant and risk of severe retinopathy among patients with diabetes duration <25 years. The Lys505 variant of glycoprotein Ia was associated with an odds ratio for severe retinopathy of 1.88 (95% confidence interval 0.83 to 4.24). Overall, there was a significant interaction between glycoprotein Ia genotype and duration of diabetes on the risk of retinopathy (P-value for interaction = 0.019). These results suggest the hypothesis that genetic variation of platelet glycoprotein Ia may play a particularly important role during the advanced stages of diabetic retinopathy. PMID- 12540965 TI - Stabilization of monocyte chemoattractant protein-1-mRNA by activated protein C. AB - The activated protein C (APC) pathway has been suggested to be a common link between coagulation and inflammation. APC may function to restore hemostasis via modulation of cytokine expression. We investigated the effect of APC on the endothelial expression of monocyte chemoattractant protein-1 (MCP-1), a chemokine that is controlled by the activation of central proinflammatory transcription factors, such as nuclear factor-kappa B (NF-kappaB). We found that human APC (2.5 10 micro g/ml) upregulated the amount of MCP-1-mRNA in human umbilical vein endothelial cells (HUVEC) and caused a time- and dose-dependent increase in MCP-1 protein production (p <0.001 for APC 2.5 micro g/ml at 4 up to 24 h). In this cell culture model MCP-1 induced an improvement of cell migration and wound repair after injury to endothelial monolayers. After stimulation of MCP-1-mRNA transcription with TNF-alpha (0.1-1 ng/ml), HUVEC's were washed and an inhibitor of gene transcription, Actinomycin D (1 micro g/ml), was added in the presence or absence of APC. HUVEC's receiving APC contained more MCP-1-mRNA than controls after one hour and up to eight hours suggesting an inhibitory effect of APC on MCP-1-mRNA degradation (with APC: 753 +/- 56 atto mol of MCP-1-mRNA per ml of cell lysate vs. 263 +/- 60 atto mol/ml without APC at t =4 h; p <0.001). Electrophoretic mobility shift assays revealed that APC attenuated NF-kappaB DNA binding capacity implying that NF-kappaB may not be involved in the upregulatory effect of APC on MCP-1 production. The ability of APC to upregulate the production of MCP-1, most likely by increasing the stability of MCP-1-mRNA rather than by transcriptional activation via NF-B, identifies a novel immunomodulatory pathway, by which APC may control the local inflammatory reaction, thereby initiating wound repair and modulating the extent of endothelial injury. PMID- 12540966 TI - Insulin resistance: an atherothrombotic syndrome. The Leeds family study. AB - The insulin resistance syndrome (IRS) is a clustering of atherothrombotic traits associated with increased vascular risk. We investigated the degree to which the phenotypic correlations between these traits are due to shared genetic and environmental factors. A multivariate genetic analysis was performed in 537 adults from 89 healthy white north European families. All traits showed significant heritability. BMI had significant genetic correlations with fasting insulin, systolic blood pressure (sBP), plasminogen activator activator inhibitor 1 (PAI-1) and fibrinogen and triglyceride. Fasting insulin had a significant genetic correlation with fibrinogen and triglyceride and Factor VII (FVII). Significant genetic correlations were shown between triglyceride and PAI-1, fibrinogen and FVII. PAI-1 and tissue plasminogen activator (t-PA) showed significant genetic correlation with sBP and with each other. Pleiotropy was demonstrated between fibrinogen and PAI-1, t-PA and FVII. Significant environmental correlations were also demonstrated. This study demonstrates pleiotropy between coagulation and fibrinolytic factors. Shared genetic and environmental factors influencing haemostatic, metabolic and anthropometric traits underlie the atherothrombotic nature of the IRS. PMID- 12540967 TI - Vascular endothelial growth factor enhances venous thrombus recanalisation and organisation. AB - Vascular endothelial growth factor (VEGF) is a regulator of physiological and pathological angiogenesis and is found in naturally resolving experimental venous thrombi, where it may also regulate recanalisation. In this study VEGF protein was injected into venous thrombi to determine if this enhanced recanalisation and organisation. A rat model of inferior vena cava (IVC) thrombosis was used. Thrombi were formed in 3 groups (n = 3 per group). 10 micro l (125)I-VEGF was directly injected into thrombus thirty minutes after induction. Three hours, 1 day and 6 days later thrombus, IVC, and other tissues were harvested. (125)I-VEGF was mostly distributed in the thrombus and the IVC, with smaller amounts in other tissues. Thrombi were formed in a further 4 groups (n = 6 per group). Thirty minutes after induction control solution or 1 ng, 10 ng or 100 ng recombinant human VEGF(165) was injected directly into the thrombus. Lumen recanalisation, thrombus organisation and monocyte content were measured on digitised sections by image analysis. In animals treated with 10 ng of VEGF there was a greater area of lumen recanalisation [mean 5492 pixels, standard error of mean (sem) 922] compared to controls (mean 2974, sem 385) (P = 0.005). There was a significant increase in the organisation score in all treated animals (1 ng: mean 70, sem 1.7, P = 0.0025; 10 ng: mean 70, sem 2.0, P = 0.0042; 100 ng: mean 72, sem 1.9, P = 0.0003) compared to controls (mean 63, sem 1.7). The monocyte content was lower in the animals treated with 1 ng VEGF (mean 3.8% of thrombus area, sem 0.3%) compared to controls (mean 5.5%, sem 0.4%) (P = 0.0008). The proportion of monocytes migrating to the centre of the thrombus increased in a dose-related manner. VEGF may prove to be of use in the treatment of venous thrombosis. PMID- 12540968 TI - Vascular endothelial growth factor (VEGF-A) plasma levels in non-small cell lung cancer: relationship with coagulation and platelet activation markers. AB - Platelet activation, commonly found in lung cancer patients, may cause the release of angiogenic factors, such as vascular endothelial growth factor (VEGF A). The present study was designed to investigate whether plasma VEGF-A levels were associated to different stages of non-small cell lung cancer (NSCLC). Moreover, sP-selectin, prothrombin fragment 1+2 (F1+2), thrombin-antithrombin III complex (TATc) and D-dimer levels were measured to test the hypothesis of an involvement of platelet and coagulation activation in tumor angiogenesis. VEGF-A, sP-selectin, F1+2, TATc and D-dimer levels were elevated in 65 patients with NSCLC, particularly in metastatic patients. sP-selectin (p <0.003) and F1+2 (p <0.005) levels were independently associated to VEGF-A. In addition, patients with positive levels of both sP-selectin and F1+2 had the highest levels of VEGF A. In conclusion, our findings support the hypothesis that thrombin generation might induce platelet activation and VEGF-A release in NSCLC. PMID- 12540969 TI - Correlates of thrombin generation in patients with advanced prostate cancer. AB - Thrombin generation is increased in men with advanced prostate cancer. Thrombin has the ability to interact with, and affect the biology of, a variety of cell types including prostate cancer cell lines. We therefore looked for correlations between thrombin generation and other markers of disease activity in spot urine samples obtained from men with advanced prostate cancer. Excretion of part of the prothrombin activation peptide F(1+2) (called here iF2), interleukin-6 (IL-6), the bone turnover marker deoxypyridinoline (DpD), and vascular endothelial growth factor (VEGF) were quantitated in spot urine samples collected from 37 men with hormone-refractory prostate cancer. Following log transformation of the data, significant correlations were found by univariate analysis between the excretion of a marker of thrombin generation (iF2) and IL-6, DpD and VEGF, as well as between IL-6 and DpD or VEGF excretion. No correlation was found between any marker and serum PSA level. After multivariate analysis, a significant correlation remained between thrombin generation and IL-6 excretion. Analysis of a second urine specimen obtained from 19 of the subjects 1 to 7 months after the first also revealed a significant correlation between thrombin generation and IL 6, DpD, and VEGF excretion. These data provide evidence of a correlation between thrombin generation/coagulation system activation and IL-6 generation in patients with cancer. They provide a rationale for studying the effects of inhibitors of thrombin generation upon the biology of prostate cancer. PMID- 12540970 TI - Angiogenesis: molecular mechanisms and functional interactions--2nd Kloster Seeon Meeting of the German Priority Research Grant "Angiogenesis". AB - The German Priority Research Program "Angiogenesis" (www.angiogenese.de) hosts a biannual meeting in the Kloster Seeon in Southern Germany. The 2nd Kloster Seeon Meeting "Angiogenesis: Molecular Mechanisms and Functional Interactions" was held in September 2002. It included sessions on hypoxia, the biology of endothelial precursor cells, angiogenic growth factors including VEGFs, the angiopoietins, ephrins, and FGFs, mechanisms of vascular sprouting and cell-cell contacts during angiogenesis, angiogenic signaling, lymphangiogenesis, angiogenesis during tumor invasion and metastasis, and on novel angiomanipulatory therapies. This report summarizes the key findings reported during the platform presentations of the meeting. PMID- 12540971 TI - In patients symptomatic for deep-vein thrombosis factor VIII elevation is found twice as frequent as in patients symptomatic for pulmonary embolism. PMID- 12540973 TI - [Treatment of superficial thrombophlebitis]. AB - Superficial thrombophlebitis is a frequently occurring disease which, because of risk factors, affects women to a greater extent than men. The presence of deep venous thrombosis should be excluded by imaging procedures. Heparin or low molecular weight heparins are indicated for treatment, if deep venous thrombosis is present or threatening. In the acute stage non-steroidal antiinflammatory drugs can be taken orally in order to alleviate pain and inflammation. In the case of concomitant chronic venous insufficiency, edema protective drugs (e. g. preparations containing horse chestnut extract and flavonoids) can support the therapy. For the treatment of inflammatory conditions in relatively short superficial vein segments without involvement of the deep venous system, a limitation to topical treatment is possible. In addition to compression therapy, the application of creams or gels containing heparin or heparinoids are the usual treatment approach. The use of topical products containing mucopolysaccharide polysulfate (MPS) has been proven to be beneficial. The problem of providing evidence of the efficacy of topical products in the therapy for superficial thrombophlebitis under the current strict demands of our regulatory authorities and of developing new products under these conditions is discussed. PMID- 12540974 TI - [Diagnosis and therapy of thrombophlebitis with special consideration of low molecular weight heparin]. AB - The clinical diagnosis of superficial thrombophlebitis is characterized by the occurrence of painful and inflamed cords along superficial veins and varices. Duplex sonography is recommended to rule out asymptomatic deep vein thrombosis which may be present in about 20% and to check potential sources of entrance of the thrombotic process into deep veins, like the junctions of great and small saphenous veins. Classic therapy is based on firm compression therapy and walking exercises. Incisions with expression of clots and anti-inflammatory drugs may reduce pain and inflammation. When phlebitis involves also the thigh and especially the proximal part of the great saphenous vein surgical ligation of the junction and local thrombectomy can be considered, preferably on an outpatient basis. Recent data from one randomised controlled trial demonstrate the efficacy of unfractionated heparin in a dose of 12,500 I.U. s. c. twice a day in this indication. According to another randomised controlled trial low molecular weight heparin (LMWH) may reduce the development of thromboembolic complications and also the relatively frequent extension of the thrombi in the superficial veins. Therapeutic doses seem to be more effective than prophylactic doses. While conventional therapy with compression and walking is sufficient for the majority of cases, the additional use of low molecular heparin is recommended in increased thromboembolic risk and when the thigh is involved. In the few studies available treatment time of 6-12 days is reported. More studies with special focus on indication, dosage and duration of therapy with LMWH are needed for the recommendation of clear therapeutic guidelines. PMID- 12540975 TI - [Prevention of catheter-induced thromboses by low molecular weight heparins]. AB - Infections and thromboses are the most frequent complications of central venous catheters (cvc). The use of a special material (i. e. polyurethane, silicone) and type (i. e. Hickman) small in diameter, the avoidance of any infection as well as the optimal position of the cvc tip in the lower part of the vena cava superior and a short period of catheterisation will be effective in decreasing the number of thromboses, whereas other thrombogenic factors (i. e. malignant disease), the patient's age, additional chemo- and/ or radiotherapy cannot practically be influenced. Coated cvc, oral anticoagulants, acetylsalicylic acid, and low dose urokinase, unfractionated and predominantly low molecular weight heparins (LMWH) are in use to avoid thromboses. Our outpatients with progressed gastrointestinal malignancies, port systems, and systemic chemotherapy consequently receive a prophylaxis with LMWH for at least 90 days according to the beneficial results published in three studies. Additional antitumoral effects of LMWH are presently evaluated and will favour consequent LMWH prophylaxis in patients with advanced malignancies and permanent catheters. PMID- 12540976 TI - [Catheter-related thrombosis and its prevention in infancy]. AB - Vascular catheters are the most important cause of thromboembolism in neonates. Additionally, a concomittant genetic predisposition with the well-known mutations is often present. In order to understand the pathology, knowledge about the specific condition of the newborn is important, especially concerning the altered hemostatic balance. For indications the specific situation of the very small infant is to be considered, e. g. the increased risk of cerebral bleeding in the first days of life of prematures. Therefore, careful consideration of indication and contraindications is mandatory. To avoid catheter-related thrombosis different schedules of prophylaxis are well-tried, especially low dose heparinization (sometimes as continuous infusion) is recommended for different types of vascular access. In neonates specific organ-related complications of umbilical catheters are to be considered. For the early diagnosis of catheter related thrombosis, attention should be given to its first signs. Thrombolytic therapy is worth to be considered. However, the state of the patient and the dynamic of thrombus growth must be taken into account. PMID- 12540977 TI - [Combination therapy with anticoagulants and platelet-active drugs]. AB - Platelets and the coagulation system cannot be regarded as completely separate systems. Both systems become activated upon vessel injury and multiple functional interactions exist. Thus with a combination therapy with anticoagulants and platelet-active drugs, drug interactions relevant for hemostasis must be expected. This includes a synergistic inhibition of the hemostatic system but also possible synergistic effects on bleeding complications. This concise review discusses the clinical efficacy of a combination therapy with anticoagulants and platelet-active drugs in patients with acute coronary syndromes (unstable angina, myocardial infarction). PMID- 12540978 TI - [Fractionated stereotactically guided radiotherapy for pharmacoresistant epilepsy]. AB - AIM: This prospective study evaluated the efficiency of fractionated stereotactically guided radiotherapy as a treatment of pharmacoresistant temporal lobe epilepsy. PATIENTS AND METHODS: Inclusion criteria were patients aged between 17 and 65 years with one-sided temporally located focus, without sufficient epilepsy control by antiepileptic drugs or neurosurgery. Between 1997 and 1999, two groups of six patients each were treated with 21 Gy (7 times 3 Gy) and 30 Gy (15 times 2 Gy). Study end points were seizure frequency, intensity, seizure length and neuropsychological parameters. RESULTS: All patients experienced a marked reduction in seizure frequency. The mean reduction of seizures was 37% (range 9-77%, i. e. seizures reduced from a monthly mean number of 11.75 to 7.52) at 18 months following radiation treatment and 46% (23-94%, i. e. 0.2-23 seizures per month) during the whole follow-up time. Seizure length was reduced in five out of eleven patients and intensity of seizures in seven out of eleven patients. CONCLUSION: Radiotherapy was identified as safe and effective for pharmacoresistant epilepsy since a very good reduction of seizure frequency was observed. It is no substitute for regular use of antiepileptic drugs, but means an appropriate alternative for patients with contraindication against neurosurgery or insufficient seizure reduction after neurosurgery. PMID- 12540979 TI - Multivariate analysis of prognostic factors in patients with glioblastoma. AB - BACKGROUND: To identify prognostic factors for overall survival in patients with newly diagnosed glioblastoma undergoing radiation therapy. PATIENTS AND METHODS: From January 1980 to June 2000, we treated 432 consecutive patients with glioblastoma at out institution. 17 patients were excluded from the analysis for various reasons. Mean age of the 415 patients who were included in the study was 59 years (19-81 years), Karnofsky performance status (KPS) was > or = 70 in 280 patients. 343 patients underwent resection, 72 had a biopsy. Various fractionation schemes were used (conventional fractionation, n = 112; hypofractionation, n = 94; accelerated hyperfractionation, n = 209). Survival probabilities were estimated using the method of Kaplan and Meier. Multivariate analysis was done with a Cox regression model. RESULTS: By July 2001, 406 patients had died. Medial overall survival was 8.2 months. Of ten factors considered in a proportional hazards model stratified for treatment (fractionation scheme and type of surgery), significant variables in a multivariate model were age (50-64 years vs < 50 years [RR 1.35; 95% CI 1.02 1.78], > or = 65 years vs < 50 years [RR 2.08; 95% CI 1.54-2.81]), performance status (KPS < 70 vs > or = 70 [RR 1.53; 95% CI 1.23-1.90]), and central tumor location (yes vs no [RR 1.39; 95% CI 1.04-1.87]). Blood hemoglobin (Hb) values were available in 318 patients and serum lactate dehydrogenase (LDH) levels in 234 patients. 89 patients were anemic (Hb men < 13 g/dl, women < 12 g/dl), in 80 patients the LDH level was raised beyond the upper limit of the normal range (> 240 U/l). By including the three significant variables, both parameters had an additional significant effect with an estimated relative risk of about 1.4 in their corresponding subgroups. CONCLUSION: Besides established prognostic factors, anemia and raised serum LDH levels may negatively influence outcome in glioblastoma patients. Our results from data-dependent modeling have to be confirmed by independent studies. PMID- 12540980 TI - Recursive partitioning analysis (RPA) class does not predict survival in patients with four or more brain metastases. AB - BACKGROUND: We evaluated prognostic factors for survival in patients with four or more brain metastases in order to determine whether intense local treatment might be justified for some of them. If up to three brain metastases are present, surgical resection or radiosurgery are currently being considered in case of favorable prognostic factors. PATIENTS AND METHODS: Retrospective intention-to treat analysis of 113 patients who underwent whole-brain radiotherapy without surgical resection or radiosurgery at a single institution. Standard treatment was given with ten fractions of 3 Gy. Higher total doses were administered in 13% of patients. Recursive partitioning analysis (RPA) prognostic classes have been described by the Radiation Therapy Oncology Group (RTOG) in 1997 (class I: Karnofsky performance status [KPS] > or = 70%, age < or = 65 years, no extracranial metastases, controlled primary tumor; class III: KPS < 70%; class II: others). RESULTS: Median number of brain metastases was six (four to 50). Most patients (69%) had extracranial metastases as well. Criteria of RPA Class I (II) were met in 4% (41%), whereas 56% had KPS < 70% and thus were grouped into class III (Tables 1 and 2). Complete or partial remission of brain metastases was found in 46% of patients who underwent computed tomography. Median survival was 4 months, 1-years survival rate 15%. Only age was a borderline significant prognostic factor in univariate analysis (< or = 50 years vs > 50 years, p = 0.05). Strong trends were found for KPS, extracranial metastases, control of the primary tumor, and breast primary tumor. Number of brain metastases, RPA class and treatment-related factors such as total dose or remission of brain metastases had no appreciable influence on survival (Figure 1). Multivariate analysis failed to identify any significant prognostic factor. CONCLUSIONS: Patients with four or more brain metastases seem to represent a group with unfavorable prognosis where remission of brain metastases or administration of more than 30 Gy were not associated with increased survival. The number of patients in RPA class I was too small to draw final conclusions. However, there was absolutely no survival difference between patients in class II (median survival 3.6 months) and III (median 4.2 months). PMID- 12540981 TI - The impact of gross tumor volume (GTV) and clinical target volume (CTV) definition on the total accuracy in radiotherapy theoretical aspects and practical experiences. AB - AIM: To evaluate the impact of interobserver variability in the contouring of gross tumor volumes (GTVs) and clinical target volumes (CTVs) on the global geometric accuracy in radiation therapy. MATERIAL AND METHODS: In a review of the currently available literature, the magnitude of interobserver variability is analyzed, causes and consequences are discussed. Uncertainties due to inconsistencies in contouring are related to other sources of geometric errors, particularly patient positioning and organ motion. RESULTS: Interobserver variability is a major -- for some tumor locations probably the largest -- factor contributing to geometric inaccuracy. Causes are multifactorial and include image and observer-related factors, such as the subjective interpretation of image information. CONCLUSION: Consequences to reduce interobserver variability are proposed, among others the selection of adequate imaging modalities, intensified radiologic training, and the use of telecommunication tools. PMID- 12540982 TI - [Retrospective results of radiation therapy of the eustachian tube in chronic otitis media]. AB - BACKGROUND: The treatment results of symptomatic radiation therapy of the Eustachian tube in chronic otitis media had to be evaluated retrospectively. PATIENTS AND METHODS: Between 1980 and 1997, 66 patients were referred for therapy. The median age was 58 years. In the clinical presentation, all the patients had a hearing impairment, 35 patients complained of pain, 21 had otorrhea. In their history, 20 patients indicated chronic recurrent infections. The complaints lasted for 4.7 years in the median, primary conservative (adstringentia, antibiotics) and surgical treatment (paracentesis, tympanic tubule, tympanoplastic) did not lead to lasting cure. In 40 of 66 patients, finally radiation therapy was done of both Eustachian tubes. With opposed fields and cobalt-60 photons a total dose of 6 Gy at single doses of 1 Gy, three times a week, was applied. Under the causes for exclusion of radiation therapy were non acceptance of the patients (nine), prior radiation therapies (six) or spontaneous improvement after initial presentation in our department. The treatment results were evaluated by interviews of the patients and regular otorhinolaryngological examinations. RESULTS: There were no side effects noticed. 28 of 40 (70%) patients reported a significant improvement that could be verified by objective otorhinolaryngological examinations. In the group of 26 nonirradiated patients, 22 could be interviewed indicating in 16 cases (72%) that the complaints were unchanged and chronic otitis media was lasting. In a subgroup analysis concerning the duration of otitis media radiation therapy proved more effective in an acute and subacute stadium of disease of up to 5 years duration, while the patients resistant to radiation therapy were entirely in a chronic stage of disease exceeding 5 years duration. CONCLUSIONS: Radiation therapy is an effective tool for symptomatic improvement of the therapy-resistant chronic otitis media. A dose of 6 Gy seems to be sufficient to achieve an antiinflammatory effect. Radiotherapy should be applied earlier after initial conservative and surgical treatment. PMID- 12540983 TI - [Time factor and repopulation during fractionated radiotherapy. Comparison between two xenografted human squamous cell carcinoma]. AB - BACKGROUND: A series of experiments were performed to determine the local tumour control of two human squamous cell carcinoma lines in nude mice. An accelerated fractionated radiation therapy regime is compared to a conventional-fractionated therapy regime. MATERIAL AND METHODS: KB is a well established human nasopharyngeal squamous cell carcinoma line (ATCC CCL 17). In nude mice KB grows as an low differentiated carcinoma. PEC MB is an undifferentiated squamous cell carcinoma of the maxillary sinus, which was successfully established in nude mice by our group 1993. Both tumors were serially passaged in nude mice. Local irradiation was given without anaesthesia under ambient conditions to air breathing animals using 18 MeV electrons of an linear accelerator (Mevatron 77, Siemens, Munich). Each dose level group consists of six to eight animals. The radiation treatments were given in ten equals fractions using graded dose levels of 2, 3, 4.5, 6 and 8 Gy. The interfraction time interval was 6 hours in the accelerated-fractionated group and 24 hours in the conventional-fractionated group (Figure 1). In the conventional-fractionated group a therapy break was given after 5 fractions for 72 h. The endpoint of the experiments was the dose, which was necessary to control 50% of the tumors (TCD(50)). The TCD(50) values were calculated after 60 days (Tables 1a and 1b). RESULTS: The experiments show, that with increasing overall treatment time of 8 3/4 days using the same number of fractions under ambient conditions the tumor control dose of the tumor KB increases from 36.3 Gy (95% CI 30.9...42.7) to 44.3 Gy (38.3...51.2) (Figure 2a). For the tumor PEC MB the tumor control dose increases from 39.5 Gy (33.4...46.7) to 45.5 Gy (37.0...56.0 (Figure 2b). CONCLUSION: This observed increase of the dose necessary to control the squamous cell carcinoma KB and PEC MB can be caused by repopulation of clonogenic tumors cells, however, other mechanism such as an increasing fraction of hypoxic tumor cells can not be ruled out. PMID- 12540984 TI - Influence of fractionated irradiation on neutrophilic granulocyte function. AB - BACKGROUND: A recent study has demonstrated that radiation therapy with single doses of up to 32 Gy has only a minor effect on neutrophilic granulocyte function. In clinical practice, by contrast, fractionated irradiation is applied. Therefore, the aim of the current study was to verify the influence of fractionated radiation therapy on granulocyte function. MATERIAL AND METHODS: Density gradient-purified granulocytes of voluntary healthy donors were used for all experiments. Granulocytes were kept in RPMI 1640 without fetal calf serum, incubated for 48 h and irradiated. Their function was assessed by measuring luminol-enhanced chemiluminescence after stimulation with phorbol myristate acid (PMA). All tests were performed at least five times. RESULTS: Relative changes (any reactive oxygen species [ROS] release before stimulation was defined as being equal to 100%) in ROS release increased after stimulation wit PMA (mean +/- SD): 0 Gy: 785 +/-, 462.2%; 2 Gy: 704.3 +/- 388.1%; 6 Gy: 1,360.3 +/- 710.5%; 12 Gy: 1,119.4 +/- 581.1%; 18 Gy: 1,087.3 +/- 622.4; 6 Gy (3 x 2 Gy): 279.4 +/- 201.1%; 12 Gy (6 x 2 Gy): 278.8 +/- 175.3%; 18 Gy (9 x 2 Gy): 84.2 +/- 41.5%. Comparing relative changes in ROS release after PMA stimulation, the differences between 0, 2, 6, 12, 18 Gy, and 6 Gy (3 x 2 Gy), 12 Gy (6 x 2 Gy), 18 Gy (9 x 2 Gy), and between 6 Gy (3 x 2 Gy), 12 Gy (6 x 2 Gy) and 18 Gy (9 x 2 Gy) proved to be significant (all p < 0.005). CONCLUSION: The study shows, that clinically used fractionated irradiation has an impact on granulocyte function, but contrary to common assumption, it is not to total dose itself but rather the fractionation which influences granulocyte function. This could have a major clinical impact on radiation treatment schemes especially for benign diseases or anti-inflammatory treatment. PMID- 12540985 TI - [Craniospinal irradiation -- patient positioning and verification with portal films]. AB - BACKGROUND: Implementation of an important step for quality assurance in irradiation of the craniospinal axis was made through changes in the irradiation technique. Crucial improvements in patient positioning and the possibility of taking portal films of the field junctions are described. MATERIAL AND METHODS: A box for the positioning of the head of the patient in has been developed. In combination with vacuum cushions, it is a cheap method for the individual and secure positioning of the patient in prone position. Furthermore, a method for taking portal films of the field junctions between the lateral cranial and the upper dorsal spinal field as well as between the two dorsal spinal fields is described. DISCUSSION: The use of the "Bonner Box" improves the patient positioning with respect to reproducibility, setup time and comfort. The documentation of the field junctions in craniospinal irradiation, which is described for the first time, is an important tool for quality assurance. PMID- 12540986 TI - [Radiotherapy of keloids. Patterns of care study -- results]. AB - BACKGROUND: Keloids are benign diseases of the skin. Because of the high rate of relapses after operation often postoperative irradiation is used. Treatment modalities must be collected by Patterns of Care Studies before therapy advice may be given from DEGRO. PATIENTS AND METHOD: Dates of investigations with questionnaire on mail of 250 radiotherapy institutions in Germany in 1997/2000 were collected to know therapy modalities, results and side effects. Keloids were irradiated in 101 institutions. During a time up to 35 years 1672 patients were irradiated, 880 patients had a follow-up. RESULTS: Mostly radiotherapy was applied soon postoperatively with kilovoltage radiotherapy or electrons 4-12 MeV fractionated 3-5 times a week and single doses of 2-3 Gy up to total doses of 10 20 Gy. 101 relapses were seen, the frequency of 11.4% is in the range of literature. Relapses were seen in the interval of some weeks up to 2 years after therapy. Side effects of irradiation were low, no malignant transformation was reported. CONCLUSION: Postoperative radiotherapy for keloids is applied in Germany mostly under equal conditions. This therapy offers effective relapse prophylaxis with nearly no side effects. A follow-up for 2 years is necessary to see outcome and relapses of irradiation. PMID- 12540987 TI - [Influence of the beta-blocker nebivolol on left ventricular function in patients with chronic heart failure]. AB - BACKGROUND: Sympathetic activity is a significant predictor of a poor prognosis in heart failure. beta-blockers such as carvedilol, metoprolol or bisoprolol have been shown to improve the prognosis of patients with heart failure. AIM: This pilot study examined the tolerability and effect of the new beta-blocker nebivolol on left ventricular ejection fraction in patients with chronic heart failure. PATIENTS AND METHODS: Twelve patients with an ejection fraction of 13 39% were included into a double-blind, placebo-controlled, randomized trial with nebivolol on top of a standard therapy. Exercise time, heart rate, left ventricular function, and tolerability were examined at baseline and after 3 months of orally administered nebivolol (2.5 mg and 5 mg, n = 6) or placebo (n = 6). RESULTS: Nebivolol was well tolerated and NYHA stage improved in four patients. Heart rate decreased while the maximal exercise duration and performance remained stable. Left ventricular function improved (ejection fraction: increase from 29.8 +/- 10.66% to 41.2 +/- 10.53%; p = 0.007) after 12 weeks of nebivolol whereas placebo did not improve cardiac function statistically significant. Left ventricular endsystolic diameter decreased from 56.5 +/- 9.40 mm to 50.2 +/- 9.43 mm in the nebivolol group (p < or = 0.02). CONCLUSION: These data indicate that nebivolol might improve cardiac function in patients with chronic heart failure. PMID- 12540988 TI - [Scl-90-R scoring of stress after myocardial infarction]. AB - OBJECTIVE: 134 patients with acute myocardial infarction, who were treated in an intensive care unit (ICU) of a German hospital in Kiel between 1996 and 1999 were selected for the study to examine their psychologic symptoms. PATIENTS AND METHODS: The patients were asked to fill in the self-report inventory, the Symptom Checklist-90-Revised (SCL-90-R) within the first 48 h after their arrival in the ICU. RESULTS: Patients with a low social status were found to have elevated mean values of the following scales: somatization (0.93), depression (0.66), and anxiety (0.59). The Global Severity Index (GSI) as a global measure of psychologic distress was elevated as well (0.46). CONCLUSION: Since patients with a limited social network and low social status, increased anxiety and depressive scores have a poor prognosis as to their mortality and their professional reintegration, it it considered necessary for the prospective setting of this study to register particularly those patients in order to treat their anxiety. PMID- 12540989 TI - [Stem cell transplantation in primary systemic vasculitis]. AB - BACKGROUND: Many patients with various autoimmune diseases fail to respond to conventional immunosuppressive therapy and develop irreversible organ damage. In some cases the complications might be fatal. Furthermore, prolonged immunosuppression with cyclophosphamide or corticosteroids often leads to long term side effects, cumulative organ damage, and development of secondary malignancies. THERAPY: Thus, short-term, high-dose immunosuppressive therapy with autologous stem cell transplantation might be an alternative for otherwise refractory patients. This concept has been used mainly in patients with scleroderma, systemic lupus erythematosus, rheumatoid arthritis, and multiple sclerosis. PATIENTS: Patients with primary systemic vasculitis (PSV) seem to be suitable candidates for high-dose chemotherapy (HDCT). Although complete, long term remissions are possible with standard immunosuppressive therapy, there is a subgroup of patients who cannot be cured by standard therapy. They need long-term immunosuppression and eventually die due to progression of the disease or cumulative therapy-related toxicity. In large series, the 5-year mortality rate was > 20%. The severity of vasculitis rather than the distinction between subgroups should determine whether HDCT might be a life-saving treatment. RESULTS AND CONCLUSION: Careful patient selection with the help of scoring systems and determination of the optimal time in the course of disease are now the major goals in the approach to HDCT. First reports as well as our own single-center experience in HDCT with a limited number of patients with severe PSV have shown that long-term remissions are possible even in patients refractory to conventional immunosuppression. PMID- 12540990 TI - [Hypersensitivity vasculitis]. AB - BACKGROUND: Hypersensitivity vasculitis (leukocytoclastic vasculitis) is defined as small-vessel vasculitis mediated by deposition of immune complexes (Arthus reaction) after exposition to various agents, such as drugs, toxins and infections. Due to a wide spectrum of precipitating agents and symptoms, classification systems and synonyms, understanding of the disease, its diagnosis and therapeutic strategy are difficult. METHODS: Based on an extensive analysis of the literature, causal agents, etiopathogenesis as well as symptoms and their frequencies are summarized. The widespread differential diagnoses and the development of a diagnostic strategy for practical management of the disease are discussed. RESULTS AND CONCLUSIONS: The course of disease is generally benign, and spontaneous remission is often observed. Severe organ manifestations and chronic courses have, however, been described. Clinical diagnosis must be confirmed histopathologically. Once the diagnosis is made, search for etiologic agents (successful in only 50% of the cases) and an intensive organ diagnostics should be performed. Subsequently, treatment with steroids, antihistamines and slow-acting antirheumatic drugs such as cytotoxic substances and immunosuppressive drugs is planned according to severity and prognosis. PMID- 12540991 TI - [Prognostic factors in thin cutaneous malignant melanoma]. AB - An increasing number of thin (< or = 1 mm) cutaneous malignant melanomas are currently diagnosed. The majority of thin lesions is associated with an excellent prognosis, however, some of them may develop local recurrences and/or distant metastases with fatal outcome. Although Breslow thickness is the single most significant prognostic factor in melanoma in general, for thin lesions the identification of other morphological, biological and/or molecular parameters which may have an impact on neoplastic progression is mandatory. At present, Clark's levels and ulceration are regarded as significant prognostic factors. Further studies are needed to confirm the prognostic value of other histopathological parameters, including the evaluation of regression, inflammatory infiltrate and mitotic activity. PMID- 12540992 TI - [The Ljubljana classification of epithelial hyperplastic laryngeal lesions: a morphometric evaluation of nuclear pleomorphism]. AB - Epithelial hyperplastic laryngeal lesions (EHLL) are associated, with a varying degree of "epithelial risk"- to develop invasive carcinoma. Several classifications have been proposed but none has received a total agreement. The 1999 Ljubljana classification distinguished four grades: simple, abnormal and atypical hyperplasia and in situ carcinoma (ISC). The first two grades are considered benign lesions; the ISC is the malignant lesion, while the atypical hyperplasia is considered a "risky lesion". This is characterized by alterations of epithelial cells towards malignancy, but not to the extent to be found in carcinoma cells. Such characteristics refer to cytomorphological (e.g., nuclear hyperchromatism, nucleoli, increased nuclear/cytoplasmic ratio) and architectural (e.g. stratification, orientation, maturation) features. In the Ljubljana scheme, nuclear pleomorphism is one of the most important features. We wanted to improve the importance of nuclear pleomorphism in the basal cells layer in different classes of EHLL using morphometrical analysis. We studied 8 cases of simple hyperplasia, 10 of abnormal hyperplasia, 10 of atypical hyperplasia and 8 of ISC using the software SAM (Shape Analytical Morphometry). The results were submitted to univariate statistical analysis. Nuclear dimensions (maximum diameter, perimeter and area) showed a progressive increase from simple to atypical hyperplasias to ISC, while abnormal hyperplasia showed the lowest values. On the contrary, analytical parameters related to nuclear contour irregularities and asymmetries showed their highest values in abnormal hyperplasia nuclei. There were no significant differences between atypical hyperplasia and ISC, while it was possible to differentiate abnormal hyperplasia from the others. In conclusion basal nuclei of atypical hyperplasia and ISC are similar so that other cytological and morphological architectural parameters are necessary to distinguish the two lesions. Abnormal hyperplasia seems to be the biological category of 'proliferative " benign laryngeal epithelium; simple hyperplasia refers to "stable" - irritative epithelium. PMID- 12540994 TI - [Uterus in a male: persistent Mullerian duct syndrome. Report of a case with embryological considerations]. AB - We describe a case of persistent mullerian duct syndrome incidentally found in a 79-year-old man. It was characterized by the presence of a uterus-like retroperitoneal mass composed of an ovoidal corpus and rudimental portio with a central lumen. Histologically the mass was leiomyomatous in nature, closely resembling myometrium, while the central lumen was lined by a monolayer of ciliated cuboidal to columnar epithelium similar to embryo-fetal celomatic epithelium. Although this is a dysembryogenetic syndrome due to the lack of regression of the mullerian duct, it may be incidentally discovered in elderly patients, as in the present case during hernioplastic surgery. We also briefly discuss the regression of mullerian duct, focusing on morphological features observed in a large collection of human embryos and fetuses, in order to better understand the pathogenesis of a "uterus' in a male without evidence of sexual abnormalities. PMID- 12540993 TI - [Breast calcifications with percutaneous vacuum-assisted biopsy diagnosis of malignancy or atypical hyerplasia: correlations with surgical findings]. AB - Percutaneous, stereotactic, vacuum-assisted biopsy has become a widely used alternative to open surgical biopsy for the initial diagnosis of breast calcifications. We retrospectively assessed the accuracy of the technique in the diagnoses of malignancy and atypical hyperplasia by correlation with the findings of the subsequent surgical excision. We studied 330 consecutive cases of breast calcifications, 216 (65.5%) of which were determined to be benign and 114 (34.5%) to be malignant or atypical at vacuum-assisted biopsy using an 11 gauge instrument. Of the latter 93 were available for comparison with the subsequent surgery, the specific diagnoses as revealed by percutaneous biopsy were as follows: 11 cases of atypical ductal hyperplasia (ADN), 67 cases of ductal carcinoma in situ (DCIS), 6 infiltrating ductal carcinomas (IFDC), 2 cases of atypical lobular hyperplasia and 7 of lobular carcinoma in situ (LCIS). At histological analysis after surgical excision, 3 (27%) of 11 cases previously diagnosed as ADH and 6 (9%) of 67 cases diagnosed as DCIS were shown to actually be higher grade lesions (DCIS/IFDC and IFDC, respectively). Of the 7 lesions diagnosed at vacuum-assisted biopsy as LCIS, surgery and histological analysis showed one infiltrating globular carcinoma and two DCIS. A total of 21 lesions (4 ADH, 14 DCIS, 1 IFDC, 2 LCIS) were completely removed at percutaneous biopsy; the remaining cases were found totally concordant. These data Indicate a substantial accuracy of the percutaneous biopsy: some lesions (particularly those thought to be ADH and DCIS) can be underestimated for sampling error. PMID- 12540995 TI - [Dermatofibrosarcoma protuberans of the breast: a case report]. AB - We describe a dermatofibrosarcoma protuberans (DFSP) of the breast and briefly report about such cases previously mentioned in the literature. A 27-year-old woman was referred because of the progressive increase, during pregnancy, of a mammary nodule located between the internal quadrants of the right breast. Its clinical and radiologic features suggested a fibroadenoma. Lumpectomy revealed a 3-cm, gray-whitish, fasciculated nodule. Histological examination showed a neoplasm characterized by a highly and monomorphic cellular proliferation of spindle-shaped cells, arranged in bundles displaying repetitive storiform growth pattern and infiltrating the adjacent mammary tissue. Lack of necrosis and low mitotic rate was observed. At immunohistochemistry the tumor cells were diffusely positive for vimentin and CD34, but negatively stained with CD99, bc-2, desmin, smooth-muscle actin, S100 protein and cytokeratins. A diagnosis of mammary dermatofibrosarcoma protuberans (DF-SP) was posed. Neoplastic involvement of surgical margins led to a subsequent quadrantectomy without regional lymphadenectomy. The patient was alive and disease-free at the 8-month follow-up. Mammary DFSP is rare. Its preoperative diagnosis is extremely difficult, particularly when radiologic images show an intraparenchymal lesion with round borders. We report such a case and briefly review the pertinent literature. Morphologic parameters to distinguish DFSP from other spindle cell lesions of the breast are discussed. PMID- 12540996 TI - Primary Hodgkin's disease of the nasopharynx: a rare but bona fide disease. AB - We report an uncommon case of Hodgkin's disease confined to the nasopharynx. An isolated polypoid mass of the nasopharynx was observed in a 41-year-old man presenting with increasing bilateral nasal obstruction. Histological study revealed a mixed cellularity type of Hodgkin's disease. Immunohistochemical analysis revealed CD30 and LMP1 expression and a lack of reactivity to CD15, CD3 and CD20. Serological tests excluded recent infection with Epstein-Barr virus. HD of nasopharynx is rare, but has to be recognized as such in view of appropriate treatment. PMID- 12540997 TI - Collagenous spherulosis of the breast in association with in situ carcinoma. AB - Collagenous spherulosis of the breast is distinctive breast histological finding characterised by the accumulation of basement membrane material in form of eosinophilic or rarely basophilic spherules that exhibit concentric and radiating fibrillar patterns. It is usually an incidental microscopic uni- or multifocal finding that occurs adjacent to benign proliferative lesions, but it can be also coexistent with malignant breast processes. Its occurrence within a malignant process is exceptional. We present a case of collagenous spherulosis of the breast associated with poorly differentiated carcinoma in situ in a 54-year-old woman. This rare change can be misinterpreted as adenoid cystic carcinoma. PMID- 12540998 TI - [A new technique for foetal brain fixation and extraction]. AB - We performed a new technique for foetal brain fixation and extraction that offers a remarkable reduction in sampling time facilitates specimen handling while conserving high quality. With the new fixation method, it is possible to obtain samples adequate for macroscopic and microscopic observations and immunohistochemical analysis. The technique involves the creation of an ex vacuo phenomenon in the subarachnoideal space prior to injecting the fixative solution. In this manner, the solution is homogenously distributed. Ease, reproducibility and the possibility of standardizing the procedure are the principal advantages. Low costs, reduced wording time and less need for human resources are other advantages. Histologically, we obtained, quickly, high quality slides with routine and immunohistochemical stains. Disadvantages of the technique derive from the use of formaldehyde and glacial acetic acid, rather than water to wash the samples; thus one must work in a well ventilated area, with gloves, protective glasses and an adequate lab coat to avoid skin and respiratory tract damage. PMID- 12540999 TI - [LAT (linker for activation of T cells): a useful marker for megakaryocyte evaluation on bone marrow biopsies]. AB - Detection of atypical megakaryocytes in bone marrow biopsies, especially in cases of myelodysplastic syndromes (MDS), chronic myeloproliferative disorders (CMPD) and acute leukemias, is facilitated by staining for markers such as Ulex europaeus agglutinin (UEA)-J, CD31, CD61 and von Willebrand factor (VWF), the latter being considered the most sensitive. Recently, LAT (linker for activation of T cells), a molecule involved in T-cell activation and platelet aggregation, was found to be expressed by megakaryocytes and platelets in tissue sections. We compared VWF and LAT immunoreactivity on megakaryocytes in 64 bone marrow biopsies from 12 normal controls (NC), and from patients with MDS (n=18), CMPD (n=21) and acute megakaryocytic leukemia (AML-M7, n=13). Immunostaining was performed on paraffin sections with polyclonal antibodies against VWF and LAT. Immunoreactivity was evaluated by counting positive megakaryocytes in 10 high power fields, and values were compared using Student's t test for paired data. Both VWF and LAT predominantly stained the cytoplasm of megakaryocytes, although LAT was also recognizable on the cell membrane. In most biopsies, the immunoreactivity of the two antibodies was quite similar. No significant differences were noticed between the mean values of VWF+ and LAT+ megakaryocytes. However, in 22 cases (5 NC; 5 MDS; 6 CMPD; 6 AML-M7), the number of LAT+ megakaryocytes was at least 30% higher than VWF+cells, while in 3 cases opposite findings were found. In 3 AML-M7 cases, anti-LAT antibodies stained numerous megakaryocytes, but anti-VWF staining was practically negative; in another 5 AML M7 cases, anti-LAT labeling was much stronger than anti-VWF staining. LAT represents a useful immunohistochemical marker for megakaryocytes in normal and pathological conditions. It seems to be expressed by megakaryocytes more than VWF in most cases and, particularly, in conditions associated with poorly differentiated megakaryocytes, such as acute megakaryocytic leukemias. The use of LAT staining should be recommended in association with other megakaryocyte markers in the study of bone marrow biopsies in cases of hematopoietic disorders. PMID- 12541000 TI - Predictive factors in prostate needle biopsy. AB - Prostate cancer (PCa) is a leading cause of morbidity and mortality in men, accounting for approximately 30% of all new cases of cancer and approximately 14% of cancer deaths. Despite considerable advances achieved in the ability to detect and treat PCa, there have not been significant corresponding decreases in PCa related morbidity and mortality. Proper examination of prostate biopsy specimens by pathologists is critical in determining the type of treatment and predicting patient outcome. The goal is to tailor the therapeutic approach to the clinical, morphological and molecular features of each patient. PMID- 12541001 TI - ADME evaluation in drug discovery. 1. Applications of genetic algorithms to the prediction of blood-brain partitioning of a large set of drugs. AB - In this study, the relationships between the brain-blood concentration ratio of 96 structurally diverse compounds with a large number of structurally derived descriptors were investigated. The linear models were based on molecular descriptors that can be calculated for any compound simply from a knowledge of its molecular structure. The linear correlation coefficients of the models were optimized by genetic algorithms (GAs), and the descriptors used in the linear models were automatically selected from 27 structurally derived descriptors. The GA optimizations resulted in a group of linear models with three or four molecular descriptors with good statistical significance. The change of descriptor use as the evolution proceeds demonstrates that the octane/water partition coefficient and the partial negative solvent-accessible surface area multiplied by the negative charge are crucial to brain-blood barrier permeability. Moreover, we found that the predictions using multiple QSPR models from GA optimization gave quite good results in spite of the diversity of structures, which was better than the predictions using the best single model. The predictions for the two external sets with 37 diverse compounds using multiple QSPR models indicate that the best linear models with four descriptors are sufficiently effective for predictive use. Considering the ease of computation of the descriptors, the linear models may be used as general utilities to screen the blood-brain barrier partitioning of drugs in a high throughput fashion. PMID- 12541002 TI - The prevalence of autism spectrum disorders. Recent evidence and future challenges. AB - BACKGROUND: Until recently best estimate prevalence rates for autism spectrum disorders (ASD) were 0.5/1,000 for autism and 2.0/1,000 for the broader spectrum. Three recent studies have suggested a significantly higher prevalence rate for ASD of 6.0/1,000 (mean 95 % CI = 4.8-8.0). METHOD: Possible determinants of the apparent increase in the prevalence of ASD are outlined. Methodological aspects of the three recent studies are examined. FINDINGS: Increased recognition, the broadening of the diagnostic concept over time and methodological differences across studies may account for most or all of the apparent increase in prevalence, although this cannot be quantified. CONCLUSIONS: Findings from ongoing studies should help confirm or disconfirm the putative rate of 6.0/1,000 for all ASD. The possibility that autism has been over-diagnosed in recent studies needs to be ruled out. Notwithstanding these outstanding questions, it appears likely that the current true prevalence of ASD is considerably greater than previously recognised. This has significant implications for our scientific understanding of ASD and for families and services. Future directions for epidemiological research are outlined. PMID- 12541003 TI - Mental and physical health of out of school children in a South African township. AB - BACKGROUND: Poor school attendance has negative consequences for the individual and society. We investigated physical and mental health correlates of school attendance amongst black children in Cape Town, South Africa. METHODS: We undertook a cross-sectional community study of children, aged 6-16 years, living in Khayelitsha township. Multistage sampling produced a sample of 499 children. They, and/or their parents, were interviewed using a Xhosa translation of the Diagnostic Interview Schedule for Children Version 2.3 (DISC-2.3). Prevalence of mental and physical health problems of school attenders were compared with those of non-attenders. RESULTS: 23.6 % of children were not attending school. Young children from unserviced areas were at particular risk. Male school attenders suffered higher levels of chronic illness than non-attenders. No differences in the prevalences of psychiatric disorders were found. CONCLUSIONS: There is no significant excess of physical or mental health problems in out of school children. Poverty plays an important role in failure to attend school. PMID- 12541004 TI - Obsessive compulsive behaviour and depressive symptoms in young people with Tourette syndrome. A controlled study. AB - Tourette syndrome (TS) is characterised by multiple motor and one or more vocal tics. There have been no controlled studies using standardised instruments of depressive symptoms and obsessive compulsive symptomatology (OCS) in young people with TS. We completed a study of phenomenology and psychopathology in children with TS, including a controlled evaluation of the association between depressive symptoms, OCS, and TS. 57 people aged 15 or under with TS were recruited. Phenomenology and psychopathology were assessed using standardised instruments. The association between TS, depressive symptoms and obsessionality was investigated using 75 age- and gender-matched controls. There were high levels of depressive symptomatology and OCS in the TS group. Twenty-three (40 %) had carried out self-injurious behaviours and 34 (60 %) met criteria for Attention Deficit Hyperactivity Disorder (ADHD). Depressive symptoms and obsessionality were higher in the TS cohort compared with the control group; this excess persisted after adjustment for the effects of age, gender and comorbidity between depression and obsessionality. This study demonstrates high levels of psychopathology in children with TS, including ADHD, OCS and depressive symptoms. The findings illustrate the potentially complex, challenging combination of difficulties encountered by children with TS and those who care for them. PMID- 12541005 TI - A pilot randomised control trial of a parent training intervention for pre-school children with autism. Preliminary findings and methodological challenges. AB - Few attempts have been made to conduct randomised control trials (RCTs) of interventions for pre-school children with autism. We report findings of a pilot RCT for a parent training intervention with a focus on the development of joint attention skills and joint action routines. Twenty-four children meeting ICD-10 criteria for childhood autism (mean age = 23 months) were identified using the CHAT screen and randomised to the parent training group or to local services only. A follow-up was conducted 12 months later (mean age = 35 months). There was some evidence that the parent training group made more progress in language development than the local services group. However, the present pilot study was compromised by several factors: a reliance on parental report to measure language, non-matching of the groups on initial IQ, and a lack of systematic checking regarding the implementation of the parent training intervention. Furthermore, three parents in the local services group commenced intensive, home based behavioural intervention during the course of the study. The difficulties encountered in the conduct of RCTs for pre-school children with autism are discussed. Methodological challenges and strategies for future well-designed RCTs for autism interventions are highlighted. PMID- 12541006 TI - Anxiety and fear. Discriminant validity in the child and adolescent practitioner's perspective. AB - OBJECTIVE: We assessed the ability of child and adolescent practitioners to discriminate between anxiety items from the Revised Children's Manifest Anxiety Scale (RCMAS) and fear items from the Fear Survey Schedule for Children-Revised (FSSC-R). In addition, we examined the effects age, gender, nationality, and therapeutic orientation on discrimination ability. METHOD: Child and adolescent psychiatrists and psychologists from two university hospitals in Australia and the USA completed a questionnaire comprised of items randomly chosen from the RCMAS and the FSSC-R. Clinicians rated each item on the extent to which the item represented the construct of anxiety or fear, using a 7-point Likert-type scale. RESULTS: Clinicians were more accurate in their perceptions of anxiety than in their perceptions of fear. Clinicians with a psychodynamic orientation were more likely to perceive an item as describing anxiety, and were less likely to identify fear. There was a significant interaction between age, scale and perception, with the youngest clinicians showing the greatest perceptual differentiation between the fear and anxiety items. CONCLUSIONS: The results suggest a need to develop common terminology among researchers and clinicians, develop scales with items specific to the pathology they intend to measure, and consider the variables influencing the clinicians rating them. PMID- 12541007 TI - Predictors of length of stay in inpatient child and adolescent psychiatry: failure to validate an evidence-based model. AB - OBJECTIVE: To test if predictors of length of stay are based on empirical evidence. METHOD: 1001 regularly terminated treatment episodes from 13 child and adolescent psychiatric hospitals were analysed. In order to cross- validate the results, the sample was randomly divided into a definition sample (n = 500) and a validation sample (n = 501). The variables in the definition sample were screened statistically for their suitability as predictors of logarithmic length of stay (logLOS). Variables shown to be significant and uncorrelated were entered into multifactor analyses of variance in order to generate the model with the largest amount of explained variance of logLOS. Subsequently the results were tested against the validation sample. RESULTS: In the definition sample we found the three predictor variables admission as crisis intervention, out of home dispositions and psychoanalytic therapy which could explain 23.7 % of the variance of logLOS. Unfortunately, this could not be replicated in the validation sample as a model. CONCLUSION: Simple models of prediction of LOS in the field of child and adolescent psychiatry cannot be reliably based on empirical evidence. The main consequence is that fixed disorder-related reimbursement systems do not seem justified. PMID- 12541008 TI - Personality disorders in former child psychiatric patients. AB - The present case-control study was undertaken in order to investigate the long term outcome with respect to personality disorder (PD) symptomatology in former child psychiatric in-patients as compared to matched controls from the general population. Altogether 359 former patients and 359 controls were invited to participate in the study. Of these, 164 (46 %) former patients and 193 (54 %) controls approved participation. From these, 137 age and sex-matched pairs with a mean age of 30.7 (SD = 6.8) years were constructed. Adult PD symptomatology was assessed by means of the DSM-IV and ICD-10 Personality Questionnaire (DIP-Q). There were 52 former patients (38 %) and 15 controls (10.9 %) who fulfilled criteria for at least one DSM-IV self-reported PD. There was a significantly higher prevalence for all specific self-reported PDs in former patients compared to controls. The mean number of disorders was 1.7 (SD = 2.6) in former patients and 0.3 (SD = 0.8) in controls. Moreover, former patients fulfilled more PD criteria than controls (23 vs. 11; median numbers). The former patients had significantly lower global functioning and more psychosocial problems than the controls. These problems were related to personality pathology. The results of this study indicate that child psychiatric morbidity seems to increase the risk for adult PD symptomatology. However, the results may be biased by the low participation rate. PMID- 12541009 TI - Kynurenate production by cultured human astrocytes. AB - In the rodent brain, astrocytes are known to be the primary source of kynurenate (KYNA), an endogenous antagonist of both the glycine(B) and the alpha7 nicotinic acetylcholine receptor. In the present study, primary human astrocytes were used to examine the characteristics and regulation of de novo KYNA synthesis in vitro. To this end, cells were exposed to KYNA's bioprecursor L-kynurenine, and newly formed KYNA was recovered from the extracellular milieu. The production of KYNA was stereospecific and rose with increasing L-kynurenine concentrations, reaching a plateau in the high microM range. In an analogous experiment, astrocytes also readily produced and liberated the potent, specific glycine(B) receptor antagonist 7-chlorokynurenate from L-4-chlorokynurenine. KYNA synthesis was dose dependently reduced by L-leucine or L-phenylalanine, two amino acids that compete with L-kynurenine for cellular uptake, and by aminooxyacetate, a non-specific aminotransferase inhibitor. In contrast, KYNA formation was stimulated by 5 mM pyruvate or oxaloacetate, which act as co-substrates of the transamination reaction. Aglycemic or depolarizing (50 mM KCl or 100 microM veratridine) conditions had no effect on KYNA synthesis. Subsequent studies using tissue homogenate showed that both known cerebral kynurenine aminotransferases (KAT I and KAT II) are present in astrocytes, but that KAT II appears to be singularly responsible for KYNA formation under physiological conditions. Taken together with previous results, these data suggest that very similar mechanisms control KYNA synthesis in the rodent and in the human brain. These regulatory events are likely to influence the neuromodulatory effects of astrocyte-derived KYNA in the normal and diseased human brain. PMID- 12541010 TI - Intracerebroventricular administration of betacytotoxics alters expression of brain monoamine transporter genes. AB - Intracerebroventricular (icv) administration of betacytotoxics alters brain monoamine neurotransmission, without producing hyperglycemia. By means of in situ hybridization, we have investigated the expression of dopamine, noradrenaline and serotonin transporter (DAT, NAT, 5-HTT, respectively) mRNAs, in the brain of alloxan- and streptozotcin-icv treated rats. DAT1 mRNA expression is increased in 1-week alloxan-icv treated rats in arcuate nucleus (+51%) and ventral medial bundle /VMB/ (+32%), and decrease in VMB of 4-week alloxan- (-53%) and streptozotocin- (-9%) icv treated rats, respectively. NAT1 mRNA expression in locus coeruleus is decreased in 4-week alloxan-icv treated rats (-35%) and increased in A1 cell group of 1- (+19%) and 4- (+14%) week streptozotocin-icv treated rats. 5-HTT mRNA expression in dorsal raphe nucleus is increased in 1- (+13%) and 4- (+21%) week alloxan-icv treated rats. Observed changes may suggest altered response to antidepressants in streptozotocin-icv treated rats, used as an animal model of sporadic Alzheimer's disease. PMID- 12541011 TI - Effect of peripheral lipopolysaccharide injection on dopamine content in murine anterior olfactory nucleus. AB - Norepinephrine turnover rate in the murine locus coeruleus (LC) is known to be enhanced by the intraperitoneal (i.p.) injection of lipopolysaccharide (LPS). Approximately 40% of LC neurons are also known to project to the olfactory bulb (OB) and the anterior olfactory nucleus (AON). Therefore, we investigated whether an i.p. injection of 500 microg LPS could modulate the catecholamine biosynthesis in these sites in 8-week-old C3H/HeN male mice. Unexpectedly, the content of norepinephrine was not elevated in both sites during 6-h-observation after LPS injection. The contents of dopamine and its metabolites in the AON were highly increased at 4 h after LPS injection, whereas those in the OB were not elevated during 6-h-observation. Although the AON has been considered not to belong to the dopaminergic neuron system, our report is the first to show an elevated dopamine content in the AON under a stressful condition such as endotoxemia. PMID- 12541012 TI - Neurochemical development of the brainstem catecholaminergic cell groups in rat. AB - The postnatal development of tyrosine hydroxylase activity has been studied in the brainstem catecholaminergic cell groups (A1C1, A2C2, A5, A6, A7), involved in cardiorespiratory control. In rat, at birth and at postnatal days P3, P7, P14, P21 ant P68, we used a microdissection technique followed by in vivo measurement of the tyrosine hydroxylase (TH) activity, the rate-limiting enzyme in catecholamine synthesis. There is two successive marked increases in TH activity: at P3 in every catecholaminergic cell groups (A1C1, +225%; A2C2, +300%; A5, +190%; A6, +205% compared to birth) and during the third postnatal week with a peak of TH activity at P14 (A6, +90% above the P7 level) or at P21 (A1C1, +715%; caudal A2C2, +585%; rostral A2C2, +15%; A5, +445%; A7, +180% compared to P7). The data suggest the existence of two temporal windows during the neurochemical development of the catecholaminergic cell groups, which correspond to two metabolic transitions. The first one could be related to the intra-, extrauterine transition and the second one, to a deep energetic phase of maturation in the rat brain, closely related to the maturation of cardiorespiratory processes. PMID- 12541013 TI - Polymorphisms of the alpha-synuclein promoter: expression analyses and association studies in Parkinson's disease. AB - Mutations of the alpha-synuclein gene have shown to be relevant in some rare families with autosomal dominant Parkinson's disease (PD). Furthermore, alpha synuclein protein is a major component of the Lewy bodies also in sporadic PD patients. Increased levels of wildtype alpha-synuclein in the cell leads to increased intracellular hydrogen peroxide levels and causes death of dopaminergic neurons in rat primary culture. Subsequently, oxidative stress has been directly linked with alpha-synuclein aggregation in vitro. This raises the question whether increased alpha-synuclein expression might be linked to higher susceptibility to PD and whether alpha-synuclein promoter polymorphisms are associated with PD. Here, two polymorphisms (-116C>G and -668T>C) of the alpha synuclein promoter defining four haplotypes have been characterized in 315 German PD patients. The influence of the four haplotypes on gene expression was studied by CAT reporter gene assays in neuronal SK-N-AS cells. The -668C/-116G haplotype revealed significant higher CAT expression than the -668T/-116G or the -668T/ 116C haplotype, respectively. Although the -668C/-116G haplotype was more common in PD patients, this difference was not significant. PMID- 12541014 TI - Cortical glucose metabolism is altered in aged transgenic Tg2576 mice that demonstrate Alzheimer plaque pathology. AB - Alzheimer's disease is associated with markedly impaired cerebral glucose metabolism as detected by reduced cortical desoxyglucose utilization, by altered activities of key glycolytic enzymes or by reduced densities of cortical glucose transporter subtypes. To determine whether formation and/or deposition of beta amyloid plays a role in the pathology of glucose metabolism, transgenic Tg2576 mice that overexpress the Swedish mutation of the human amyloid precursor protein and demonstrate a progressive, age-related cortical and hippocampal deposition of beta-amyloid plaques, were used to study expression and activity of key enzymes of brain glycolysis (phosphofructokinase, PFK) and glyconeogenesis (fructose1,6 bisphosphatase; FbPase). Quantitative RT-PCR revealed high expression levels of both C- and M-type PFK mRNA in non-transgenic mouse cerebral cortex, whilst there was little expression of the L-type. In 24-month-old transgenic Tg2576 mouse cortex, but not in 7-, 13-, and 17-month-old mice, the copy number of PFK-C mRNA was significantly reduced in comparison to non-transgenic littermates, while the mRNA level of the other PFK isoforms and FbPase did not differ between transgenic and non-transgenic tissue samples. In situ hybridization in brain sections from aged Tg2576 mice revealed reduced PFK-C mRNA expression in beta-amyloid plaque associated neurons and upregulation in reactive astrocytes surrounding beta amyloid deposits. The decreased PFK-C protein level detected by Western analysis in cerebral cortical tissue from 24-month-old transgenic Tg2576 mice was accompanied by reduced enzyme activity of PFK in comparison to non-transgenic littermates. Our data demonstrate that impairment of cerebral cortical glucose metabolism occurs only due to the long-lasting high beta-amyloid burden. This results from a reduction in glycolytic activity in beta-amyloid plaque-associated neurons and a concomitant upregulation in reactive, plaque-surrounding astrocytes. PMID- 12541015 TI - The role of diet in cognitive decline. AB - Recent findings suggest a possible role of diet in age-related cognitive decline, and cognitive impairment of both degenerative (Alzheimer's disease, AD) or vascular origin. In particular, in an older population of Southern Italy with a typical Mediterranean diet, high monounsaturated fatty acids energy intake appeared to be associated with a high protection against cognitive decline. In addition, dietary fat and energy in older people seem to be risk factors, while fish consumption and cereals are found to reduce the prevalence of AD in the European and North American countries. Moreover, foods with large amounts of aluminium-containing additives or aluminium from drinking water may affect the risk of developing AD. Vitamin deficiencies, especially vitamin B6, B12 and folates, and antioxidant deficiencies (vitamins E and C) could also influence the memory capabilities and have an effect on cognitive decline. Dietary anti oxidants and supplements and specific macronutrients of the diet may act synergistically with other protective factors opening new possibilities of intervention for cognitive decline. PMID- 12541016 TI - Spectrum of binge eating symptomatology in patients treated with clozapine and olanzapine. AB - The authors explored the binge eating symptomatology in 74 patients receiving clozapine (N = 57) or olanzapine (N = 17), and compared body mass index (BMI, kg/m(2)) and weight gain in patients with and without binge eating symptomatology. Subjects who screened positively for binge eating were interviewed using a modified version of the Questionnaire on Eating and Weight Patterns (QEWP). Current BMIs were assessed cross-sectionally, BMIs at initiation of clozapine/olanzapine treatment retrospectively. Thirty-seven subjects (50%) screened positively. Taking clozapine and olanzapine together, 6/27 (22.2%) females and 3/47 (6.4%) males fulfilled criteria for binge eating disorder, 3/27 (11.1%) females and 2/47 (4.3%) males for bulimia nervosa. Patients who screened positively showed higher current BMIs (26.8 +/- 3.9 vs. 24.7 +/- 3.7 kg/m(2)) and higher BMI increments during clozapine/olanzapine treatment (3.9 +/- 3.1 vs. 2.6 +/- 3.4 kg/m(2)) than patients who screened negatively. We conclude that clozapine/olanzapine may induce binge eating and full blown eating disorders which may have predictive value for weight gain. For future research in this field we suggest a novel DSM-IV research classification "Medication-induced eating disorders". PMID- 12541017 TI - Is a bilateral modified radical neck dissection beneficial for patients with papillary thyroid cancer? AB - PURPOSE: We previously reported a poor prognosis in papillary thyroid carcinoma patients with recurrence in the contralateral cervical lymph nodes after thyroidectomy and ipsilateral modified radical neck dissection (MRND) with a curative intent. The aim of this study was to investigate whether bilateral MRND improved prognosis in patients at risk for contralateral nodal recurrence. METHODS: We retrospectively reviewed the cases of 86 patients with contralateral nodal metastasis who underwent a thyroidectomy with bilateral MRND (group 1) and the cases of 32 patients who suffered contralateral nodal recurrence after a thyroidectomy with ipsilateral MRND (group 2). RESULTS: Although tumor progression was considered similar in patients from groups 1 and 2, group 1 developed fewer distant metastases than group 2 (6.8% vs 31.1%), thus resulting in a lower cancer death rate (5.8% vs 28.1%). Group 1 had a better 10-year survival rate than of group 2 (97.1% vs 83.7%). CONCLUSION: Bilateral MRND during initial surgical management is thus considered to improve the prognosis of some papillary carcinoma patients at risk for recurrence in the contralateral cervical lymph nodes. PMID- 12541018 TI - Conservative treatment for small intestinal intussusception associated with Henoch-Schonlein's purpura. AB - PURPOSE: Emergency laparotomy is generally considered the appropriate course of action for small bowel intussusception associated with Henoch-Schonlein's purpura (HSP). In this paper, we define a conservative approach after witnessing spontaneous reduction of ileoileal invagination at laparotomy in a patient with HSP who had been on steroid therapy for renal involvement. METHODS: HSP was diagnosed by the appearance of a purpuric rash without thrombocytopenia. Intussusception was diagnosed by ultrasonography (USG) and plain abdominographs, which showed signs of obstruction, and clinical examination. Barium enema was used to treat ileocolic intussusceptions, and conservative therapy, consisting of nasogastric drainage, steroids, and intravenous fluid administration, was used to treat ileoileal intussusceptions. Emergency laparotomy was performed for the patients unresponsive to therapy within 24 h, those with peritonitis, and those with ileocolic invagination not able to be reduced by barium enema. RESULTS: Six children with an ileoileal intussusception and one with an ileocecal intussusception were studied. The average age was 6 years old. Apart from the initial patient in whom spontaneous reduction was seen at laparotomy, three others required emergency laparotomy; for ileocolic intussusception unable to be reduced by barium enema in one, for ileoileal invagination with peritonitis on admission in one, and for ileoileal intussusception unresponsive to conservative therapy in one. The other three patients were successfully treated by conservative therapy. CONCLUSION: Conservative therapy is feasible for HSP patients with small bowel intussusception as long as the time of onset is known, an ultrasonographic and X-ray diagnosis is confirmed, emergency operating facilities are available, and an experienced pediatric surgical team follows up the patients. PMID- 12541019 TI - Management of adrenal metastasis from hepatocellular carcinoma. AB - PURPOSE: Although the adrenal gland is a common site of extrahepatic metastasis from hepatocellular carcinoma (HCC), there are no definitive guidelines for the treatment of adrenal metastasis. This study examines the effectiveness of various treatments for this disease. METHODS: We retrospectively analyzed 20 patients treated for adrenal metastasis of HCC by adrenalectomy ( n = 13), transarterial chemoembolization (TACE), or percutaneous ethanol injection therapy (PEIT) ( n = 7). RESULTS: There were no significant differences in cumulative survival rates between patients given adrenalectomy and those given TACE or PEIT, either after completing treatment for primary HCC or after the first treatment for adrenal metastasis. Six of seven patients with tumor thrombi in the inferior vena cava (IVC) from adrenal metastasis underwent adrenalectomy combined with intracaval thrombectomy, five of whom survived for more than 1 year after surgery, and two of whom are still alive without any recurrence more than 3 years after surgery. PEIT showed good results for small adrenal metastasis. CONCLUSION: These findings suggest that therapeutic modalities should be chosen according to the clinical features of each individual, including the size of the metastatic tumor, whether there is invasion into the IVC, the function of the remaining liver, and the existence of intra- and/or nonadrenal extrahepatic lesions. Furthermore, intracaval tumor thrombectomy could be indicated for patients with IVC thrombus if they are suitable candidates for surgery. PMID- 12541020 TI - Vascular reconstruction in Buerger's disease: is it feasible? AB - PURPOSE: Thromboangiitis obliterans (Buerger's disease) is a clinical syndrome characterized by segmental occlusions of the distal vessels. Although a cessation of using nicotine products usually helps, nevertheless a surgical revascularization may be needed in cases of stage III and IV limbs. Because of the distal and segmental nature of the disease, these procedures are rarely feasible. This article focuses on the feasibility of performing a vascular reconstruction in thromboangiitis obliterans. METHODS: Thirty-six of 94 patients (38.3%) who were followed by the Peripheral Vascular Unit of Istanbul Medical Faculty were selected for revascularization and 27 of 36 (81%) patients underwent revascularization procedures. RESULTS: During a 36-month follow-up, the patency rates at the 12th, 24th, and 36th months were 59.2%, 48%, and 33.3%, respectively. The limb salvage rate was 92.5%. CONCLUSIONS: Since patients affected by Buerger's disease consist a group of young population who are still in their productive stages, every effort should be taken to obtain a limb salvage in the ischemic period. Although the patency rates do not seem promising, the limb salvation rate was quite satisfactory. PMID- 12541021 TI - Changes in gastrointestinal motility after subtotal colectomy in dogs. AB - PURPOSE: This study was conducted to examine recovery from diarrhea after subtotal colectomy by observing changes in gastrointestinal motility, neuropeptides, and autonomic nerves in the enteric nervous system in dogs. METHODS: The moisture volume of intestinal matter recovered from the ileal side of the anastomosis was defined as the diarrhea index. Gastrointestinal motility was measured using strain gauge force transducers (SG) for 24 weeks after subtotal colectomy. Immunohistochemical changes in neuropeptides in the residual intestine, acetylcholinesterase activation of cholinergic fibers, and fluorohistochemical changes of adrenergic fibers in the myenteric plexuses were evaluated. RESULTS: The digestive tract transit time returned to normal in the early postoperative period, but decreased moisture volume was seen in the intestinal matter after 8 postoperative weeks. Migrating contraction waves propagating beyond the site of anastomosis, signaling recovery of intestinal motility, were noted after 8 postoperative weeks, corresponding to reconstruction of cholinergic fibers in the anastomotic section. Transient activation of vasoactive intestinal polypeptide was seen in the residual intestine, but this had normalized by the eighth postoperative week. Lengthened villi in the residual intestine were also seen after 8 postoperative weeks. CONCLUSION: The decreased moisture volume in the intestinal matter is due to the improvements we observed in the eighth postoperative week. PMID- 12541022 TI - Dielectric spectrogram for evaluating ischemic microstructural changes of the liver in simple cold preservation. AB - PURPOSE: Monitoring ischemic liver injury is important in liver transplantation. We previously reported that dielectric properties were correlated with tissue adenosine triphosphate during liver preservation; however, it is unknown what the changes in dielectric properties are based on. Using rats, we studied the relationship between dielectric parameters and microstructural changes in liver tissues during preservation. METHODS: We serially measured the percent decrease in conductivity and the percent increase in relative permittivity in rat livers, and determined the areas of cellular and extracellular components using a charge coupled device microscope and an NIH image. Thereafter, we evaluated the relationship between dielectric parameters and microstructural changes in the liver tissues. RESULTS: A positive linear and exponential correlation was found between the percent decrease in conductivity and the percent decrease in extracellular components ( P < 0.01). A positive linear and exponential correlation was also found between the percent increase in relative permittivity and the percent increase in cellular components ( P < 0.01). CONCLUSIONS: Our results suggest that the serial changes in tissue conductivity and the relative permittivity reflect changes in liver tissue microstructures during cold preservation. PMID- 12541023 TI - Thyroid tuberculosis mimicking carcinoma: report of two cases. AB - Among 527 patients with thyroid disease who underwent surgery at our hospital during a 20-year period, 2 (0.4%) had tuberculous thyroiditis mimicking carcinoma. The first patient was a 44-year-old man with a solitary thyroid nodule and the second was a 24-year old man with a thyroid abscess. The unexpected diagnosis was made postoperatively and was based on histological findings in both patients. No primary focus was found elsewhere in either patient, and both responded to antituberculous chemotherapy. Although the diagnosis is usually based on examination of resected specimens, recent reports indicate that find needle aspiration cytology is a cost-effective technique of diagnosing thyroid tuberculosis. A review of 35 cases reported in the English literature is also discussed. PMID- 12541024 TI - Bilateral cervicomediastinal neurofibroma originating from the vagal nerve in a patient with von Recklinghausen's disease: report of a case. AB - A 19-year-old woman with von Recklinghausen's disease was admitted with symptoms of hoarseness. A computed tomography scan showed a bilateral cervicomediastinal tumor. An extirpation of the left cervicomediastinal tumor was performed for the purpose of diagnosis and treatment. On thoracotomy, the tumor, which measured 9 x 8 x 4 cm in size, arose from the intrathoracic vagal nerve and the left tumor was resected with a segment of the vagal nerve and recurrent nerve. The pathological diagnosis of the tumor was a neurofibroma. The tumor on the right side was left untreated due to concerns about possibly causing palsy of the bilateral recurrent nerve and also because of the asymptomatic state of the right tumor. Mediastinal neurofibroma in a patient with von Recklinghausen's disease often arises from the intrathoracic vagal nerve. To our knowledge, this is the first report of bilateral cervicomediastinal neurofibroma originating from the vagal nerves. PMID- 12541025 TI - A massive hiatal hernia that mimics a congenital diaphragmatic hernia. An unusual presentation of hiatal hernia in childhood: report of a case. AB - A massive hiatal hernia containing the colon, intestine, and stomach with organoaxial volvulus is an uncommon entity in childhood. This clinical form of a hiatal hernia may mimic congenital diaphragmatic hernia and chest pathologies. In this paper, we describe a patient who presented with a massive hiatal hernia that mimicked a congenital diaphragmatic hernia, and discuss the pitfalls in diagnosing this clinical entity. PMID- 12541026 TI - Irinotecan plus low-dose cisplatin for alpha-fetoprotein-producing gastric carcinoma with multiple liver metastases: report of two cases. AB - Alpha-fetoprotein (AFP)-producing gastric carcinoma generally causes multiple liver metastases and has an extremely poor prognosis. There is no standard chemotherapy for this disease. Two recent consecutive patients who had AFP producing gastric carcinoma were treated with a novel chemotherapy regimen: irinotecan hydrochloride (100 mg/body over 90 min) plus low-dose cisplatin (10 mg/body) by intravenous infusion. Treatment was done weekly during admission and once every 2 weeks on an outpatient basis. Both patients had multiple liver metastases with high serum levels of AFP, and one demonstrated resistance to 5 fluorouracil. In both patients, liver metastases showed a dramatic complete response to chemotherapy, and the serum AFP levels returned to normal. No significant toxicities were observed. These preliminary results suggest that the present regimen may cause fewer side effects while retaining its synergistic antitumor activity. This regimen may therefore be worth trying as first-line chemotherapy for patients with metastatic AFP-producing gastric carcinoma. PMID- 12541027 TI - Solitary splenic metastasis from gastric cancer: report of a case. AB - We report a rare case of metachronous and solitary metastasis to the spleen from gastric cancer. A 69-year-old man who had undergone a distal gastrectomy for gastric cancer 48 months earlier was found to have a solitary lesion in the spleen, and an increased serum carcinoembryonic antigen (CEA) level. The patient underwent a laparotomy for suspected metastasis to the spleen. At laparotomy, a tumor was found in the upper pole of the spleen without involvement of other organs, and a splenectomy was performed. Histological examination confirmed that the splenic tumor was a well-differentiated adenocarcinoma similar to the primary gastric cancer. The postoperative course was uneventful and his serum CEA decreased to within normal levels. The patient died of multiple metastases to the liver and peritoneal dissemination 40 months after the splenectomy. PMID- 12541028 TI - Perforated duodenal ulcer associated with an incarcerated hiatal hernia: report of a case. AB - We report the case of a perforated duodenal ulcer and diffuse peritonitis associated with an incarcerated hiatal hernia. A 77-year-old woman with a 17-year history of rheumatoid arthritis treated with nonsteroidal anti-inflammatory drugs, who had also been receiving treatment for non-Hodgkin's lymphoma over 4 years, was referred to us for investigation of nausea and vomiting. An abdominal compute tomography (CT) scan showed an incarcerated hiatal hernia and free air in the hernia sac. Emergency laparotomy revealed an incarcerated hiatal hernia involving the stomach, transverse colon, and omentum. A perforated ulcer was also found in the posterior wall of the first portion of the duodenum. The combination of these disorders is thought to be rare in patients with a hiatal hernia and free air in its sac. As the reported mortality of perforated gastric ulcer associated with a hiatal hernia is high, early elective surgery should be performed in patients with a duodenal ulcer associated with a hiatal hernia. PMID- 12541029 TI - Ileal perforation caused by cytomegalovirus infection in a patient with recurrent gastric cancer: report of a case. AB - We report a case of ileal perforation caused by cytomegalovirus (CMV) infection in a patient with peritoneal recurrence of gastric cancer. Emergency laparotomy revealed a pinhole-sized perforation in a reddish segment of the small bowel, 100 cm proximal to the terminal ileum, and peritoneal carcinosis of recurrent gastric cancer invading the transverse colon and the gastrojejunal anastomosis of a Billroth-II procedure. The affected ileum was resected, a primary anastomosis was performed, and a colostomy was made in the ascending colon. The histology of the ileum revealed acute inflammation with vasculitis and CMV inclusions in the macrophages and endothelial cells and evidence of CMV on immunostaining. There was no evidence of cancer cell invasion or any other pathogens. Although the prognosis associated with bowel perforation due to CMV infection is poor, emergency surgery saved our patient's life. PMID- 12541030 TI - Primary malignant fibrous histiocytoma of the small intestine presenting as an intussusception: report of a case. AB - Malignant fibrous histiocytoma occurs most commonly in the extremities and trunk, but rarely in the visceral organs. Malignant fibrous histiocytoma of the small intestine is an extremely rare condition. To our knowledge, only ten cases of primary tumors involving the small intestine have been described up to now. Among them only one case has been reported to lead to intussusception. This case report documents the appearance of a storiform-pleomorphic type primary malignant fibrous histiocytoma of the ileum, which led to intussusception, in a 58-year-old man with a 3-month history of dyspepsia, weight loss, general fatigue, and nonspecific abdominal pain. The patient was well with no sign of disease at 8 months after surgery. The clinical and pathologic characteristics of malignant fibrous histiocytoma of the small intestine as found in the literature are also reviewed. PMID- 12541031 TI - Transmesosigmoid hernia: report of a case. AB - Sigmoid mesocolon hernia is an uncommon type of internal hernia. A 63-year-old man who presented with pain in the left side of the abdomen and nausea was referred to our department for treatment of ileus. He was initially managed conservatively, but as his symptoms became progressively worse, a laparoscopy was done, which revealed hemorrhagic ascites and necrosis of the small intestine in the lower abdomen. An open laparotomy was subsequently performed and the intraoperative findings were consistent with a transmesosigmoid hernia. There was an abnormal defect in the sigmoid mesocolon and protrusion of about 30 cm of small bowel through this abnormal opening, which had resulted in strangulation of the bowel. The necrosed part of the intestine was resected and the defect was closed. PMID- 12541032 TI - Solitary fibrous tumor in the retroperitoneal space: report of a case. AB - Solitary fibrous tumors (SFTs) are spindle-cell neoplasms originally described in the pleura. It is now known that these tumors can develop in many sites. This report describes the case of a well-circumscribed tumor located around the superior mesenteric artery (SMA), which was initially thought to be either a superior SMA aneurysm, a lymphoma, or a neurogenic tumor. Histological examination demonstrated the tumor to be composed of a cellular proliferation of ovoid to spindle cells with a fine collagenous matrix in the short fascicles. Immunohistochemical staining was strongly positive for CD34 and negative for factor VIII, cytokeratin, desmin, and muscle-specific actin (HHF-35). These findings suggested a diagnosis of SFT in the retroperitoneal space. To our knowledge, this is the first report of an SFT located around the SMA. Based on the above findings, it is important to include SFT in the differential diagnosis of retroperitoneal tumors located around the SMA. PMID- 12541033 TI - Surgical treatment for acute ischemia caused by bilateral popliteal artery entrapment syndrome in an elderly patient: report of a case. AB - We report the unusual case of a 73-year-old man who underwent surgery for bilateral popliteal artery entrapment syndrome (PAES). A medial approach was used to operate on the left leg, and the vein bypass was made from the superficial femoral artery to the crural artery through a subfascial route. A posterior approach was used to operate on the right leg and it was found that the mid popliteal artery passed medial to and beneath the medial head of the gastrocnemius muscle and was severely compressed by an accessory slip of muscle. The vein bypass from the above-knee to below-knee popliteal artery was established through the original route after resection of the accessory slip of muscle. A postoperative arteriogram showed good bypass flow to the bilateral crural arteries. To our knowledge, this case represents the oldest patient with this disorder to be treated by surgery. PMID- 12541034 TI - Surgical anatomy of hepatoduodenal ligament and hepatic hilus. AB - The anatomical situation and variations of structures in the hepatoduodenal ligament and hepatic hilus are a legacy of their embryological development. The vascular, biliary, and lymphatic structures contribute to the functioning of blood and bile flow as well as lymph drainage of the liver. Connective tissue, fatty tissue, and the peritoneal sheet are enveloping underlying structures. Their position, shape, and relation to neighboring structures influence the situation during operative procedures. The cystic artery origin is variable, as is the number; and its recognition is important for safe cholecystectomy. Venous drainage of the gallbladder goes into the portal system of adjacent segments 4 and 5 and influences the spread of gallbladder pathology. There are some surgically important variations in the course and distribution of bile ducts and arteries in the hepatoduodenal ligament. The biliary anatomical variations significantly influence the incidence of bile ducts injuries during laparoscopic cholecystectomy. The arterial supply of extrahepatic bile ducts is delicate and variable and should be considered when trying to prevent ischemic injuries to the bile ducts. Inflammation and the combination of inflammation and anatomical variation are thought to contribute to a dangerous situation in regard to eventual injury to the bile ducts and vascular structures during operative procedures. This paper explores these questions. PMID- 12541035 TI - Laparoscopic cholecystectomy for acute cholecystitis. AB - The application of laparoscopic cholecystectomy (Lap. C) for acute cholecystitis (AC) remains controversial from the viewpoint of its higher rate of morbidity, and conversion to open surgery, in spite of the worldwide acceptance of Lap. C as the gold standard for the treatment of patients with symptomatic gallbladder diseases. The conversion rate has been reported to decrease with experience. Local and overall complication rates were shown to correlate with the time delay between the onset of acute symptoms and the operation. Although percutaneous gallbladder drainage (PGBD) has been reported to be a safe and effective procedure for the treatment of AC, it should be limited to high-risk groups such as elderly or critically ill patients. Early cholecystectomy within 4 days from the onset is strongly recommended to minimize surgical complications and to increase the chance of a successful laparoscopic approach. PMID- 12541036 TI - Urgent versus interval laparoscopic cholecystectomy for acute cholecystitis: a comparative study. AB - BACKGROUND/PURPOSE: The surgical management of patients presenting with acute cholecystitis remains controversial. The aim of this study was to evaluate the safety and feasibility of urgent laparoscopic cholecystectomy (LC) during the "index" (acute) admission with acute cholecystitis, and to compare that with a policy of interval LC. METHODS: Between October 2000 and October 2001, 50 patients who had suffered with acute cholecystitis underwent LC. Thirty-three patients underwent surgery during the index admission (group I), of whom 11 patients had surgery within 96 h of admission. Seventeen patients were referred by colleagues to outpatients for, and underwent, an interval LC (group II). RESULTS: All operations were completed laparoscopically. There was no difference between the groups in the operating time (median [interquartile range]: 78 [61 124] versus 93 [53-128] min) or postoperative hospital stay (median, 1 day). The delay in performing an urgent LC beyond 96 h did not affect the operating time or postoperative stay but significantly increased the total hospital stay (median [interquartile range]: 5 (5-8) versus 13 [8-17] days; P = 0.001). CONCLUSIONS: Laparoscopic cholecystectomy during the index admission with acute cholecystitis can be performed safely and successfully. Earlier surgery has a beneficial impact for patients and the National Health Service. PMID- 12541037 TI - Avoidance of biliary injury during laparoscopic cholecystectomy. AB - Biliary injury during laparoscopic cholecystectomy is still a serious problem. Injury occurs as a result of technical errors or misidentification of ducts. Inexperience, inflammation, and aberrant anatomy are key risk factors. The most serious technical problem is cautery-induced injury. This problem may be avoided by use of cautery under very low power settings in the triangle of Calot. Misidentification injuries occur when the surgeon mistakes the common bile duct or an aberrant right hepatic duct for the cystic duct. This error usually occurs when the surgeon uses the "infundibular" technique to identify the cystic duct. This technique, which depends on seeing the cystic duct flare as it becomes the infundibulum, is especially prone to be misleading in the face of acute inflammation. This technique is unreliable and should not be used alone for anatomic identification of the ducts. It is preferable to use the critical view technique or to perform a cholangiogram. PMID- 12541038 TI - Laparoscopic pancreatic cystgastrostomy. AB - BACKGROUND/PURPOSE: Internal drainage of acute pancreatic pseudocysts is indicated after the first 6 weeks of pseudocyst documentation. It is also indicated for symptomatic chronic pseudocysts 6 cm or more in diameter. When a pseudocyst is in close contact with the posterior wall of the stomach, it is best drained by pseudocyst gastrostomy. METHODS: Intragastric surgical techniques were used in 18 patients with retrogastric pseudocysts. Intragastric ports were successfully placed in all patients, and the presence of pseudocysts was confirmed by needle aspiration in 17. RESULTS: The intragastric approach was successful in 14 patients. In three cases a small abdominal would 8 cm in length was needed. The reasons for such conversion included uncontrollable bleeding from the cystic wall in two cases and a cystic wall in contact with, but not adherent to, the posterior gastric wall in one. Bleeding was controlled through a gastrotomy, and cystgastrostomy was performed. The wound required for conversion was much smaller than that needed for planned open surgery. These procedures are called laparoscopy-assisted cystgastrostomy. Good short- and long-term results were obtained. One patient, in whom the cystgastrostomy was too small, developed a cyst infection and underwent reoperation. Except for this case, no recurrence was observed, and no further treatment was needed. CONCLUSIONS: This experience demonstrates that cystgastrostomy with the intragastric surgical technique is a safe, less invasive procedure for effectively draining a retrogastric pseudocyst. PMID- 12541039 TI - Laparoscopic pancreatic surgery: its indications and techniques: from the viewpoint of limiting the indications. AB - Indications in the field of pancreatic surgery should be limited considering the technical difficulties and the characteristics of pancreatic diseases. Benign or low-grade malignant tumors, including pseudocysts, islet tumors, and cystic tumors, are indications for distal pancreatectomy. Islet tumors such as insulinomas are good candidates for this procedure when they are located near splenic vessels or the main pancreatic duct and enucleation is considered inappropriate. Techniques of laparoscopic distal pancreatectomy with/without splenectomy and laparoscopy-assisted distal pancreatectomy indicated in low-grade malignant tumors such as mucinous cystadenoma are described. Insulinoma is one of the best candidates for enucleation because many of the cases are solitary and benign. The technique of enucleation is also described. PMID- 12541040 TI - Does laparoscopic surgery have a role in the treatment of gallbladder cancer? AB - BACKGROUND/PURPOSE: This study was conducted to evaluate the role of laparoscopic surgery in the treatment of gallbladder cancer. METHODS: A retrospective study was performed on 31 patients with a postoperative diagnosis of gallbladder cancer. The laparoscopic approach was initially applied to all of them. RESULTS: Ten patients had a pT1a cancer, and all underwent laparoscopic cholecystectomy without recurrence. Nine patients had pT1b lesions, and three had to be converted to an open operation. There were two recurrences. In one of the converted patients the cystic node was invaded. Seven patients had a pT2 lesion, and in four of them the operation was converted to an open procedure. Recurrences were noted in three patients. In two patients with pT3 cancer, the opera-tions were converted; both cancers recurred. Only diagnostic laparoscopy or a palliative laparoscopic procedure was performed for pT4 cancers. The median follow-up time was 17.0 months. The 5-years survival rate was 100% for patients with stage pT1a, 100% for pT1b, and 68% for pT2 lesions. CONCLUSIONS: We suggest that when a polypoid lesion of the gallbladder is found on preoperative evaluation, laparoscopic surgery may be attempted initially. During the procedure it is important to open all specimens when a polypoid lesion is present and perform a frozen section biopsy. When a pT1a lesion is found, laparoscopic cholecystectomy is sufficient; however, when a pT1b or more advanced lesion is found, the operation might be converted to a radical cholecystectomy. PMID- 12541041 TI - Advances in research and clinical practice in motor disorders of the sphincter of Oddi. AB - Perfusion manometry of the sphincter of Oddi (SO) using a pneumohydraulic capillary infusion system records phasic wave activity superimposed on basal pressure. A triple-lumen catheter allows the recording of propagation of the phasic waves. Microtransducer manometry is an alternative that permits prolonged recording of biliary pressure without the need for infusion. A cyclic change is recognized in SO motility in coordination with the migrating motor complex (MMC) of the duodenum during fasting. SO contractions occur at maximal frequency and amplitude during phase 3 of the duodenal MMC. Using two microtransducer catheters placed by duodenoscopy, a cyclic elevation of biliary pressure can be recorded in concert with phase 3. These findings indicate that human SO contractions impede bile flow. SO dysfunction causing biliary-type pain can be diagnosed by manometry. The pressure elevation during phase 3 may contribute to the development of pain in patients with biliary dyskinesia. Gastrectomy and proximal duodenal transection were proven to affect sphincter motility, as evidenced by the paradoxical response to cholecystokinin. Choledocholithiasis and hepatolithiasis are associated with low basal pressure, presumably due to repeated injury to the sphincter by passing stones. SO and biliary manometry leads to better understanding of biliary dynamics and the pathophysiology of biliary diseases. PMID- 12541042 TI - Biliary tract cancer treatment: results from the Biliary Tract Cancer Statistics Registry in Japan. AB - BACKGROUND/PURPOSE: A registry project for cancers of the biliary tract accumulated a total of 11,030 cases for 10 years. In the present study, registered cases were analyzed for information bearing on problems with the treatment of cancer of the biliary tract. The Japanese classification of lymph nodes was also considered on the basis of the results of this study. METHODS: In 11,030 cases, the site of cancer was the gallbladder in 4,774, the bile duct in 4,833, and the papilla of Vater in 1,423. Those cases were analyzed with regard to patient survival according to the stage of disease and the extent of lymph node metastasis. RESULTS: More than 11,000 cases of cancer of the biliary tract have been registered to date from 158 member institutions of the Japanese Society of Biliary Surgery. While the 5-year survival rates for stage I gallbladder cancer and cancer of the papilla were 77% and 75%, respectively, those for stage I hilar or upper bile duct cancer and middle or lower bile duct cancer were 47% and 54%. For stage II and stage III disease, the 5-year survival rates were about 50% for gallbladder cancer and 30% or higher for cancer of the papilla, while survival was only 20% to 30% for bile duct cancer, regardless of specific site. For stage IV, the 5-year survival rate was unexpectedly high, being about 10% or higher for cancers at all sites, with 19% for cancer of the papillary region being the highest. Thus, there still seem to remain surgical indications for stage IV cancers. The lymph node metastasis rate was about 40% for cancers at all sites. Changes in surgical procedures to improve the 5-year survival rate in patients with n2 metastasis or less will be needed. Noncurative resection occurred frequently for cancers at all sites, particularly in cancers of the hilar or upper bile duct, accounting for 60% of cases or more. We have to recognize that measures to reduce inadvertent noncurative resection are fundamental to the treatment of cancer. CONCLUSIONS: Considering the survival results according to specific lymph nodes involved, we concluded that the Japanese classification of lymph nodes, particularly hepatoduodenal ligament lymph nodes, should be reexamined, while another procedure to remove such lymph nodes completely should be developed. PMID- 12541043 TI - Multidetector row helical CT of the pancreas: value of three-dimensional images, two-dimensional reformations, and contrast-enhanced multiphasic imaging. AB - Multidetector row helical computed tomography (MD-CT) scanning is performed for the evaluation of pancreatic tumors. Three-phase contrast study is performed using 2.5-mm collimation, and the images are reconstructed at 1.25-mm intervals. CT angiography and pancreatic duct images using two- or three-dimensional techniques are reconstructed from the volumetric data. MD-CT can perform multiphasic scanning rapidly with an optimal temporal window. CT angiography obtained with MD-CT can delineate peripancreatic vasculature with high spatial resolution and sufficient vascular enhancement. Pancreatic duct images can provide important information in assessing pancreatic disease. MD-CT has the potential to improve detection and preoperative assessment of pancreatic tumors. PMID- 12541044 TI - Micrometastases from HBP malignancies and metastatic cancer. AB - Despite advances in early diagnosis and surgical treatment, the prognosis for patients with primary malignant tumors of the hepatobiliary tract and pancreas has not changed markedly over the last decades. Early metastatic relapse after complete resection indicates the presence of disseminated tumor cells undetectable by current tumor staging methods. Sensitive immunohistocytochemical and nucleic acid-based assays have been developed to detect single tumor cells present in lymph nodes, bone marrow, or blood. Standardization of the current occult tumor cell detection protocols are needed before "micrometastatic" tumor staging can be used in clinical practice. We present an overview of recent studies on the frequency and prognostic value of occult disseminated tumor cells in the lymph nodes, bone marrow, and blood of patients with hepatopancreatobiliary malignancies and metastatic colorectal cancer identified by immunohistocytochemistry or nucleic acid-based assays. PMID- 12541045 TI - Navigation and image-guided HBP surgery: a review and preview. AB - Image-guided surgery and navigation have resulted from convergent developments in radiology, teletransmission, and computer science. Patient selection and preoperative planning in hepatobiliary-pancreatic (HBP) surgery rely on preoperative imaging. The operative procedure is finally led by the fusion of additional information gained by the palpating hand and intraoperative ultrasound. Despite advances in reducing morbidity and mortality, decisions are often hardly quantifiable and are restricted to super-specialists in HBP surgery. New developments in computed tomography (CT) and magnetic resonance imaging (MRI) technology have led to the possibility of the volumetric prediction of liver resections. These data can be shared via telemedicine and used for simulation and training. Three-dimensional (3D) reconstructions have led to a better topologic understanding of tumor-vascular tree relations in the individual patient. With the increasing use of ablative procedures and laparoscopy, intraoperative imaging and navigation will hold increasing significance for the HBP surgeon. Flat screen monitors adjacent to the surgical field present computer-generated 3D virtual liver resection proposals which can be transferred into the real liver. The main obstacles in HBP navigation are the flexibility and mobility of the target organ. Intrahepatic and surface markers seem to be mandatory for computer-navigated surgery. The first feasibility studies are promising. PMID- 12541046 TI - Development of a bent-type tube stent for simultaneous drainage of bile and pancreatic juice. AB - BACKGROUND/PURPOSE: Endoscopic lithotomy is a useful medical procedure for treating choledocholithiasis. Although this procedure is commonly performed, complications such as pancreatitis and cholangitis are recognized as major and serious problems. The obstruction of bile and pancreatic juice flow caused by papillary edema or spasm is thought to be responsible for such complications. We have developed a new bent-type tube stent that can drain bile and pancreatic juice simultaneously. METHODS: From June to November 2001, temporary implantation of the new bent-type tube stent was performed in four patients (two women, two men; mean age, 73.3 years) with choledocholithiasis, for the drainage of bile and pancreatic juice simultaneously after endoscopic lithotomy by endoscopic sphincterotomy or endoscopic balloon sphincter dilatation. RESULTS: Immediately after the implantation of the new type of tube stent, bile and pancreatic juice flow from the respective ducts was recognized under endoscopic observation. Neither pancreatitis nor cholangitis occurred after these procedures. CONCLUSIONS: This procedure may be a helpful means to prevent pancreatitis and cholangitis after endoscopic lithotomy. PMID- 12541047 TI - Wedge resection of the portal bifurcation concomitant with left hepatectomy plus biliary reconstruction for hepatobiliary cancer. AB - BACKGROUND/PURPOSE: We report our experience performing wedge resection of the portal bifurcation and transverse suture closure in patients undergoing left hepatectomy and caudate lobectomy plus biliary reconstruction for hepatobiliary cancer. METHODS: The procedure was performed in three patients with hilar or intrahepatic cholangiocarcinoma. After confirming that tumor invasion of the portal bifurcation was not circumferential, the portal trunk and the right posterior and right anterior portal branches were isolated and clamped. Wedge resection of the portal bifurcation was performed, taking care to secure a clear surgical margin. The edges of the portal vein were approximated, using guy sutures in the dorsal and ventral edges and a temporary central guy-suture, and portal reconstruction was carried out using a continuous transverse suture. After unclamping, good portal flow was confirmed by color Doppler ultrasonography. RESULTS: The procedure was completed successfully in all three patients; the average time of portal vein occlusion was 15 min. Two patients had postoperative complications: bile leakage and wound infection, but no patient developed postoperative hepatic failure or died. The three patients are alive without recurrence at 2, 11, and 22 months after the operation. CONCLUSIONS: Wedge resection of the portal bifurcation is easier and simpler than using a venous patch or performing segmental resection. PMID- 12541048 TI - Liver resection for hepatic metastases: 15 years of experience. AB - BACKGROUND/PURPOSE: Liver metastases, especially those from primary colorectal cancers, are treatable and potentially curable. Imaging techniques such as computed tomography, magnetic resonance, and ultrasonography have advanced in recent years and led to increased sensitivity and specificity in the diagnosis of liver metastases. Liver surgery also has been revolutionized in the past two decades. Dissection along nonanatomical lines has permitted the resection of multiple lesions that previously might have been considered unresectable. METHODS: From 1986 to 2000, 181 patients underwent liver resection for hepatic metastasis from colorectal cancer. Of these, 56 patients underwent systematic anatomical major hepatic resection and 125 underwent nonanatomical limited resection. RESULTS: Operative morbidity and mortality rates were higher in patients in whom anatomical procedures were performed. The overall 5-year survival rate of the 181 patients was 39.8%. CONCLUSIONS: An aggressive surgical procedure in patients with hepatic colorectal metastases is safe, and may prolong overall survival, and therefore should be considered in all patients with metastases confined to the liver. PMID- 12541049 TI - Pylorus-preserving pancreatoduodenectomy with Billroth I type reconstruction: a viable option for pancreatic head resection. AB - BACKGROUND/PURPOSE: The aim of this study was to investigate the efficacy of pylorus-preserving pancreatoduodenectomy (PPPD) versus standard pancreatoduodenectomy (PD). METHOD: A cohort of 80 patients were studied for 5 years using prospectively gathered data. PPPD was performed in 48 (60%) patients, and the other 32 (40%) underwent a standard Whipple's operation with partial distal gastrectomy (PD). In all cases, the gastric or duodenal stump was anastomosed with the first loop of jejunum as a Billroth I-type reconstruction. RESULTS: Overall, morbidity in the cohort of patients consisted of 10 with a pancreatic fistula, 5 with postoperative hemorrhage, 5 with sepsis, 3 with delayed gastric emptying, 2 with an anastomotic leak, and 1 each with intraabdominal hematoma or myocardial infarction. Major morbidity associated with PPPD appeared in 8 of the 48 (16.7%) patients compared with 2 of 32 (6.3%) in the PD group. There were two deaths (4.2%) in the PPPD group and one (3.1%) in the PD group. The mean length of hospital stay was 14.6 days for PPPD versus 17.1 days for PD. Of the 48 patients in the PPPD group 7 (14.6%) had a hospital stay of more than 20 days versus 8 of the 32 (25%) in the PD group. CONCLUSIONS: Contrary to recent reports, in our series the PPPD patients had a shorter hospital stay; and overall, 3 of the 80 (3.75%) patients developed delayed gastric emptying, a relatively low rate. The pancreatic fistula rate was almost threefold higher in the PPPD group than in the PD group (but did not prolong the inpatient stay). This may be due to an intact antrum secreting higher quantities of gastrin. PMID- 12541050 TI - Usefulness of dynamic magnetic resonance pancreatography with secretin [correction of secretion] injection for evaluating pancreatic exocrine function after pancreatogastric reconstructive surgery. AB - BACKGROUND/PURPOSE: The usefulness of dynamic magnetic resonance pancreatography (MRP) with secretin injection as a new method of assessing pancreatic exocrine function was evaluated in patients undergoing pancreatogastrostomy. METHODS: A series of 12 patients who had undergone pancreatogastrostomy were classified into group 1 (9 patients whose pancreas was normal preoperatively) and group 2 (3 patients with a firm pancreas accompanied by distended pancreatic ducts). The pancreatic exocrine function was evaluated using the changes in maximum pancreatic duct diameter and the intensity ratio calculated by tracing the contour line on the images of pancreatic juice secretion newly occurring after dynamic MRP with prior secretin injection. RESULTS: Clear visualization of the pancreatic duct by MRP was achieved in all 12 patients, and a decrease in pancreatic duct diameter was evident postoperatively in group 2. Pancreatic juice secretion was well visualized on MRP images following secretin injection in 10 patients but not in 2 patients of group 2. Higher intensity ratios were noted in group 1 than in group 2. CONCLUSIONS: Dynamic MRP with secretion injection enables clear verification of pancreatic juice secretion on images and permits objective evaluation of pancreatic exocrine function. It therefore is a useful procedure for assessing pancreatic exocrine function after resection of the pancreas. PMID- 12541051 TI - Undifferentiated carcinoma of the common bile duct: case report and review of the literature. AB - A 78-year-old Japanese man with undifferentiated carcinoma of the common bile duct is presented. Upon gross examination, the common bile duct was found to be obstructed by a nodule measuring 10 x 10 mm. Microscopically, the nodule was ill defined and composed of atypical spindle-shaped and pleomorphic tumor cells. The spindle-shaped cells proliferated in a whirled or interlacing pattern simulating a sarcoma, and the pleomorphic tumor cells had abundant eosinophilic cytoplasm and bizarre nuclei. Histochemically, a few tumor cells contained mucosubstances stained with the alcian blue (AB) method in their cytoplasm. Immunohistochemically, the tumor cells were diffusely positive for CAM5.2 and AE1/AE3. The histological diagnosis was undifferentiated carcinoma (spindle cell carcinoma) of the common bile duct. Other than our patient, only four other cases of undifferentiated carcinoma in the extrahepatic bile duct have been reported in the literature. PMID- 12541052 TI - Minute pancreatic carcinoma with initial symptom of acute pancreatitis. AB - We experienced a case of minute pancreatic carcinoma in a 59-year-old man who complained of upper abdominal pain after drinking alcohol. Abdominal ultrasonography (US) revealed dilatation of the main pancreatic duct (MPD). Abdominal computed tomography (CT) and magnetic resonance cholangiopancreatography (MRCP) showed slight dilatation of the MPD and its obstruction near the portal vein. Endoscopic retrograde cholangiopancreatography (ERCP) demonstrated occlusion of the MPD, and cytology of aspirated pancreatic juice was negative for malignancy. With the diagnosis of benign localized obstruction of the MPD, the patient underwent surgery. There was a clear demarcation of hardness and color of the pancreas on the left margin of the superior mesenteric vein, and the caudal pancreas was hard and fibrotic. Intraoperative US revealed slight dilatation of the MPD, and the aspiration cytology result was class IV. First, segmental resection of the pancreas was performed, but pathological examination of frozen section showed neither malignancy nor stenotic lesion. An additional small portion of the proximal pancreas was resected. The specimen included a ductal carcinoma, 5 mm in diameter. Accordingly, a pylorus-preserving pancreatoduodenectomy was performed. Microscopically, the minute carcinoma had already penetrated the duct wall and infiltrated lymph vessels and veins. The patient has been under close observation at our outpatient clinic, and so far there have been no signs of recurrence. To improve the poor prognosis of pancreatic cancer, we should be alert to the occurrence of acute pancreatitis as an initial symptom. PMID- 12541053 TI - Invasive carcinoma of the pancreas derived from intraductal papillary adenocarcinoma without mucin hypersecretion but with changes in the pancreatic duct on pancreatography. AB - We present a case of invasive carcinoma of the pancreas derived from intraductal papillary adenocarcinoma without mucin hypersecretion in a 65-year-old man with a 45-year history of alcohol abuse and a 2-year follow-up of chronic pancreatitis. Two years previously, in May 1998, he was admitted for investigation of abdominal pain. Computed tomography (CT) showed diffuse dilation of the main pancreatic duct with atrophy of the pancreatic parenchyma. Endoscopic retrograde pancreatography (ERP) showed a diffusely dilated main pancreatic duct with irregular side branches in the head of the pancreas. Chronic alcoholic pancreatitis was diagnosed on the basis of the pancreatography findings. The patient was readmitted for investigation of progressive weight loss in August 2000. Serum CA19-9 levels were markedly elevated (750 U/ml) and CT showed enlargement of the head and body of the pancreas. ERP showed irregularity of the main pancreatic duct in the head of the pancreas, and the distal main pancreatic duct (which was dilated on initial ERP examination) was interrupted in the body of the pancreas. Suspected pancreatic carcinoma was diagnosed, and pylorus preserving pancreatoduodenectomy was performed. Frozen section examination of the cut end of the pancreas revealed ductal carcinoma, and total pancreatoduodenectomy with portal vein resection was performed. Histologically, the resected tumor was diagnosed as an invasive carcinoma derived from intraductal papillary adenocarcinoma without mucin hypersecretion. We recommend observing changes in the pancreatic duct on pancreatography to diagnose invasive carcinoma of the pancreas derived from intraductal papillary adenocarcinoma in a resectable state. PMID- 12541054 TI - Focal pancreatic hemorrhage mimicking pancreas carcinoma with lymph node metastasis. AB - A 67-year-old man was referred to our department because of a mass in the pancreas tail. Pancreatic tail cancer with lymph node metastasis was diagnosed, based on various radiological findings. Computed tomography (CT) revealed a slightly enhanced tumor mass around the celiac plexus, and endoscopic retrograde pancreatography (ERP) showed complete obstruction of the main pancreatic duct at the tail, although magnetic resonance imaging (MRI) demonstrated no elevation of the mean pixel value of the tumor after gadolinium (Gd) injection, and tumor markers of pancreas cancer were within normal limits. At surgery, the mass around the celiac plexus was found to be blood coagulation, and the pancreas tail tumor was found to be a focal hematoma in the pancreas. Pathologically, the hematoma was not encapsulated, and it was diagnosed as hemorrhage in the pancreas parenchyma. Both hemorrhagic lesions were suspected to have developed respectively. It is difficult to distinguish pancreas hemorrhage from carcinoma in the pancreas with chronic pancreatitis, especially when the hemorrhage is small in size and there are other extrapancreatic hemorrhagic lesions. For such diagnosis, the superiority of MRI with Gd injection is indicated. PMID- 12541055 TI - Benign nonepithelial fibroma in the pancreas. AB - This is, to our knowledge, the first report of a fibroma originating in the pancreas in which preoperative imaging findings or explicit descriptions of histopathologic observations are clearly described. A 64-year-old man was referred to our hospital for further evaluation of a pancreatic mass. Clinical examination and laboratory analysis revealed no abnormal findings, except for mild elevation of serum trypsin (550 ng/ml). Abdominal ultrasonogram, computed tomographic scan, magnetic resonance imaging, and angiography demonstrated a well demarcated circular mass in the tail of the pancreas. A laparotomy was performed on April 26, 1995. The tumor was observed to be tan-colored and smooth, and was localized in the front of the pancreas tail, suggesting that it had arisen from the pancreatic capsule. The patient underwent enucleation of the tumor. The resected tumor presented macroscopically as a fibrous nodule measuring 5.5 x 6.0 cm. The pathological diagnosis was fibroma. Immunohistochemically, spindle cells were positive for vimentin and negative for alpha-smooth muscle actin, desmin, S 100, and neuron-specific enolase. PMID- 12541056 TI - Transmission of Yersinia pestis cultures with different plasmid content from Xenopsylla cheopis to Calomys callosus. AB - Most Brazilian Yersinia pestis isolates display a typical plasmid profile composed of the three classical plasmids: pYV, pPst and pFra. However, some cultures lack at least one of these plasmids, while a few of them harbour atypical DNA bands of molecular weight ranging from 147 to 11.5 kb. To investigate whether Y. pestis displaying atypical plasmid content could be propagated among rodents in nature through flea bites, we carried out studies with fleas ( Xenopsylla cheopis) and rodents ( Calomys callosus) reared in the laboratory and five Y. pestis cultures differing in plasmid content. The results suggest that: (1) the single presence of pYV is not sufficient for the transmission of Y. pestis by fleas, (2) pPst is not essential for transmission, (3) two atypical DNA bands of molecular weight of 30 kb and >90 kb have no biological role, and (4) pFra is required for the transmission of Y. pestis by flea bites. Other studies are needed to determine whether this plasmid alone is sufficient for transmission. PMID- 12541057 TI - Immunological and histopathological studies in a rodent model infected with Leishmania infantum promastigotes or amastigotes. AB - Infection with Leishmania infantum promastigotes (group I) and amastigotes (group II) was evaluated over 32 weeks, using Syrian golden hamsters as an experimental model. Spleen cells strongly responded to the specific antigen at 12 (group I) and 16 weeks (group II) post-inoculation (p.i.) and lower stimulation index values coincided with the parasite burden peak. Western-blot analysis detected antibodies during the 1st week p.i. and the number of recognized proteins increased with the time of infection, reaching a maximum at the peak parasite burden. Histopathology revealed hypoplasia in spleen white pulp and the liver showed a periportal infiltration of inflammatory cells and small granulomas, becoming increasingly more severe as the infection developed. Both organs exhibited a secondary amyloid deposition at the end of the experiment, especially the spleen. In this study, progressive visceral disease was observed as in natural human and canine infections; however, the incubation period was longer in the promastigote than in the amastigote infection. PMID- 12541058 TI - Seasonal occurrence of actinosporeans (Myxozoa) and oligochaetes (Annelida) at a trout hatchery in Bavaria, Germany. AB - A systematic inventory of actinosporeans and oligochaetes conducted over 3 years at a trout fish farm in Bavaria, Germany, allowed the identification of 12 actinosporeans from five collective groups: four Triactinomyxon (Triactinomyxon nov. types 1-4), two Raabeia (Raabeia nov. types 1, 2), two Echinactinomyxon ( E. radiatum, Echinactinomyxon nov. type 1), two Aurantiactinomyxon ( A. pavinsis, Aurantiactinomyxon nov. type 1) and two Neoactinomyxum (Neoactinomyxum nov. types 1, 2). Nine forms are novel but can be placed within existing collective groups. All 12 forms were detected in the laboratory in aquarium water associated with farm sediment. However, only four of these could be linked with an oligochaete host. Three families of oligochaetes were identified from the sediment: Tubificidae, represented by eight species, Lumbriculidae with one species and Naididae with two species. Only tubificid oligochaetes were found to host actinosporeans. PMID- 12541059 TI - Cercarial shedding from Galba truncatula infected with Fasciola gigantica of distinct geographic origins. AB - Galba truncatula snails were experimentally infected with either of two different isolates of Fasciola gigantica, originating from Egypt or China, to determine the influence of these isolates on the characteristics of snail infections. The survival rates of G. truncatula on day 30 post-exposure were 90.0% and 60.2% in the Egyptian and Chinese groups, respectively. The frequency of cercaria-shedding snails within the Egyptian group was 79.8%, whereas in the Chinese group it was 22.4%. The parasite origin had a significant effect on the durations of the prepatent and patent periods. The mean number of cercariae shed from the Egyptian group was significantly greater than that shed from the Chinese group (a mean of 275.5 per cercaria-shedding snail compared with 29.0). These results could be explained by the fact that G. truncatula might be a natural intermediate host for F. gigantica in Egypt, and the greater adaptability of the Egyptian miracidia of F. gigantica to unusual snail hosts. These results demonstrate the influence of the geographic origin of the parasite on the success of trematodes infecting snails. PMID- 12541060 TI - A human schwannoma cell line supports the in vitro adhesion of Plasmodium falciparum infected erythrocytes to chondroitin-4-sulfate. AB - The paucity of human cell lines expressing defined receptors for the cytoadhesion of erythrocytes infected with the human malarial parasite Plasmodium falciparumhas hampered the investigation of this important virulence property. Here, we investigate a permanent cell line derived from a human, malignant schwannoma, termed HMS-97, and show that this cell line expresses chondroitin-4 sulfate as the only surface receptor to which P. falciparum-infected erythrocytes can cytoadhere. Other common receptors for parasite adhesion, including CD36, vascular cellular adhesion molecule-1 (VCAM), intercellular adhesion molecule-1 (ICAM-1), and E-selectin are absent. Thus, HMS-97 cells are a useful tool for the study of P. falciparum adhesion to chondoitin-4-sulfate, the main receptor for parasite sequestration in the placenta. As chondoitin-4-sulfate can be readily cleaved from the cells, HMS-97 cells are also an ideal system for expressing recombinant adhesion receptors and studying their function in binding assays. PMID- 12541061 TI - Naturally acquired babesiosis in a reindeer (Rangifer tarandus tarandus) herd in Great Britain. AB - A provisional diagnosis of babesiosis was made in a reindeer herd in Scotland when seven animals died during 1997 and 1998. Additional clinical cases occurred, but the animals recovered after treatment. Thirty-one reindeer from the herd were tested for the prevalence of exposure to Babesia by the indirect fluorescent antibody test using a bovine isolate of Babesia divergens that had been passaged through gerbils. Infection rates were determined by Giemsa-stained blood smears. In addition, molecular identification of the infecting Babesiasp. was undertaken using SSU rRNA gene sequence analysis. It is likely that the organism causing babesiosis in this reindeer herd is B. divergens. PMID- 12541062 TI - Nitric oxide and its target cells in cercaria of Diplostomum chromatophorum: a histochemical and immunocytochemical study. AB - In order to advance our knowledge of the nitrergic nervous system in flatworms, the patterns of the NADPH-diaphorase (NADPH-d) reaction and cGMP immunoreactivity, after stimulation with a nitric oxide donor in the presence of an inhibitor of phosphodiesterase, were investigated in cercaria of Diplostomum chromatophorum. This is the first time the presence of NADPH-d activity has been detected in a larval fluke, and the first time the presence of cGMP immunoreactivity has been detected in a flatworm. The NADPH-d reaction occurs in the ventral sucker, the hind body and the tail. cGMP immunoreactivity was detected in the muscle cells of the body and in two pairs of sensory cells at the anterior end of the body and in the middle of the furca. The sensory cells also showed 5-HT immunoreactivity. The spatial relationship between the cGMP and the 5 HT immunoreactivities was studied. By staining with TRITC-labelled phalloidin, the pattern of the muscle fibres was revealed. PMID- 12541063 TI - Regulation of tumor necrosis factor alpha and its specific receptors during Toxoplasma gondii infection in human monocytic cells. AB - The coccidian Toxoplasma gondiiis an obligate intracellular parasite which can infect all cell types. Among the cytokines elicited by the immune response to Toxoplasma, tumor necrosis factor alpha (TNF-alpha) acts synergistically with gamma interferon (IFN-gamma) and plays a major role in host cell resistance. We have previously reported that TNF-alpha production induced by IFN-gamma/LPS decreases after T. gondii infection of human myelomonocytic THP-1 cells. Here, we investigated the regulation of TNF-alpha and its specific receptors during T. gondii infection of THP-1 cells. We found that TNF-alpha production was regulated at a post-transcriptional level and that TNF receptor expression was regulated at a pretranscriptional level. The TNF receptor I shedding and the fall in TNF-alpha levels observed after T. gondii infection would thus be induced by a parasite component with serine protease activity. These findings indicate that T. gondii participates not only in controlling the cytotoxic effects of TNF-alpha during the infection process, but also in signal transduction mediated mainly by TNF receptor I. PMID- 12541064 TI - Molecular arguments for considering Hysterothylacium fabri (Nematoda: Anisakidae) a complex of sibling species. AB - The existence of sibling species is widespread among nematodes and the recognition of these has important epidemiological implications. In an attempt to establish whether this is the case for Hysterothylacium fabri, which is present in many Mediterranean fish species, we studied its genetic diversity and analysed its population structure. To do this, we used 266 fourth stage larvae of H. fabricollected from three different host species and used two different methods for characterising genetic variability: isoenzyme electrophoresis and RAPD. The four isoenzyme loci studied are polymorphic, with five or six alleles at each. Significant differences in the deviations from the Hardy-Weinberg law were detected, especially at the PGM locus. A total of 92.1% of the markers revealed by the RAPD technique were polymorphic, demonstrating a great diversity. The UPGMA dendrogram revealed the existence of four genetic groups. Values of Nei's genetic distance, gene flow and the existence of different fixed alleles, together with the deviations from the Hardy-Weinberg law detected in the isoenzyme study, suggest that H. fabri is a complex comprising at least three sibling species with little host specificity, at least in the case of the most abundant species. PMID- 12541065 TI - Cyclosporin A-mediated killing of Leishmania major by macrophages is independent of reactive nitrogen and endogenous TNF-alpha and is not inhibited by IL-10 and 13. AB - Murine macrophages were treated with various doses of cyclosporin A (CsA) to enhance the killing of Leishmania major parasites. CsA reduced the rate of infected cells from 75% in non-treated controls to less than 15% with 1 micro g CsA/ml in a dose-dependent manner. The leishmanicidal effect was also observed when CsA was added 48 h after the infection of macrophages. In contrast, FK506, another structural non-related immunosuppressive drug with antiparasitic activities, showed no effect on the ability of macrophages to kill intracellular Leishmania parasites. Since nitric oxide has been identified as a key molecule for the leishmanicidal function of macrophages, we analyzed the role of this molecule. There was no influence on the leishmanicidal effect of CsA when L- N-(1 iminoethyl)lysine, a potent and selective inhibitor of mouse inducible nitric oxide synthase, was added. Furthermore, the presence of the macrophage-inhibiting cytokines interleukin (IL)-10 and IL-13 simultaneously or prior to CsA did not inhibit leishmania killing, while both cytokines completely prevented parasite killing by macrophages activated with gamma interferon and tumor necrosis factor (TNF). CsA was fully active on macrophages from TNF-receptor p55 knockout mice arguing against autocrine activation by TNF. We therefore conclude that the antileishmanial effect of CsA is independent of effector mechanisms employed by macrophage-activating cytokines. PMID- 12541066 TI - Oxidative damage and phosphatidylserine expression of red blood cells in cattle experimentally infected with Theileria sergenti. AB - In order to elucidate the mechanism of anemia in Japanese bovine theileriosis, we investigated the oxidative alteration of red blood cells (RBCs) in cattle infected with Theileria sergenti. As an index of RBC oxidation, the levels of 2',7'-dichlorofluorescin-diacetate (DCFH) oxidation and malondialdehyde thiobarbituric acid reactive substances (MDA-TBARS), and phosphatidylserine (PS) expression accompanying anemia were examined in experimentally infected cattle. Before the development of anemia, the concentrations of DCFH oxidation and MDA TBARS were low, and PS expression on the surfaces of RBCs was hardly seen. However, during the onset of anemia, these levels began to increase remarkably in proportion to the decrease of packed cell volume and the increase of parasitemia in all infected cattle. During the serious stage of anemia, these oxidative indices reached their maximum values. Our findings indicate that oxidative damage and loss of membrane asymmetry in RBCs are related to the development of anemia in T. sergenti infection. This oxidative damage to the RBCs might play an important role in the pathogenesis of anemia in Japanese bovine theileriosis. PMID- 12541067 TI - Purification and localization of a 10 kDa calcareous corpuscle binding protein of Spirometra mansoni plerocercoid. AB - In cestode parasites, calcareous corpuscles are thought to be associated with a number of intracellular physiologies by regulating the trafficking of mineral components. We previously separated these particular components by Ficoll, and their binding proteins of 10 kDa and 35 kDa in the Spirometra mansoni plerocercoid (sparganum). In the present study, we purified a 10 kDa protein employing octyl-Sepharose CL-4B affinity chromatography. Anti-serum raised against the purified protein showed specific reactions to the calcareous corpuscles of the worm section by immunohistochemistry, and recognized the 10 kDa protein by immunoblotting. PMID- 12541068 TI - Competitive mating interactions between Schistosoma haematobium and S. intercalatum (Lower Guinea strain). AB - Schistosoma haematobium and S. intercalatum belong to the S. haematobium group of schistosomes and can hybridize in nature where they are sympatric. They are therefore able to co-infect the same human host. Hybridization and competitive mating interactions with S. haematobium have been implicated in restricting the distribution of S. intercalatum in Africa and in the remarkably rapid replacement of S. intercalatum by S. haematobium at Loum, Cameroon. Previous studies have demonstrated the greater pairing ability of S. haematobium over S. intercalatum in hamsters infected with both species simultaneously or infected first with S. intercalatum (Lower Guinea strain) and later with S. haematobium. The present study demonstrates the greater pairing ability of S. haematobiumover S. intercalatum in hamsters infected first with S. intercalatum (Lower Guinea) and later with S. haematobium, and indicates that S. intercalatumis unlikely to spread to areas where S. haematobium is already established. PMID- 12541069 TI - Effects of hypotonicity on amino acid release in adult Echinostoma caproni as determined by thin layer chromatography. AB - Thin layer chromatography (TLC) was used to analyze the amino acids in worm incubates isotonic and hypotonic to the intestinal habitat of adult Echinostoma caproni and to analyze the free pool amino acids of these trematodes after incubation. Qualitative analysis revealed the presence of histidine, lysine, alanine, and proline in all samples of incubate and worm tissue. Quantification of histidine and lysine by TLC with densitometry gave mean concentrations of 24.1 micro g histidine/g worm per ml incubate in Locke's solution and 195.0 micro g lysine/g worm per ml incubate in deioinized (DI) water. Quantification of histidine and lysine in the worm tissue gave mean weight percents of 0.0587 and 0.0263, respectively, in worms incubated in Locke's solution and 0.0229 and 0.0163, respectively, for worms incubated in DI water. Our findings suggest that E. caproni adults may leak amino acids for osmoregulation in hypotonic environments. PMID- 12541070 TI - Ionization-cluster distributions of alpha-particles in nanometric volumes of propane: measurement and calculation. AB - The probability of the formation of ionization clusters by primary alpha particles at 5.4 MeV in nanometric volumes of propane was studied experimentally and by Monte Carlo simulation, as a function of the distance between the center line of the particle beam and the center of the target volume. The volumes were of cylindrical shape, 3.7 mm in diameter and height. As the investigations were performed at gas pressures of 300 Pa and 350 Pa, the dimensions of the target volume were equivalent to 20.6 nm or 24.0 nm in a material of density 1.0 g/cm(3). The dependence of ionization-cluster formation on distance was studied up to values equivalent to about 70 nm. To validate the measurements, a Monte Carlo model was developed which allows the experimental arrangement and the interactions of alpha-particles and secondary electrons in the counter gas to be properly simulated. This model is supplemented by a mathematical formulation of cluster size formation in nanometric targets. The main results of our study are (i) that the mean ionization-cluster size in the delta-electron cloud of an alpha particle track segment, decreases as a function of the distance between the center line of the alpha-particle beam and the center of the sensitive target volume to the power of 2.6, and (ii) that the mean cluster size in critical volumes and the relative variance of mean cluster size due to delta-electrons are invariant at distances greater than about 20 nm. We could imagine that the ionization-cluster formation in nanometric volumes might in future provide the physical basis for a redefinition of radiation quality. PMID- 12541071 TI - In vivo response to the different LET regions of a 55 MeV helium ion beam. AB - We evaluated the in vivo response to heavy particle irradiation in rat tail epidermis using silver-stained nucleolar organizer regions (AgNOR) as the end point. The energy degradation of the beam across the circular section of the tail allowed us to study the damage elicited by two different LET regions of a helium beam, i.e. non-Bragg peak (NBP) and Bragg peak (BP), at different sites on the same sample. The tails were locally irradiated with a helium ion beam at different fluences. AgNOR exhibited marked variations between tissue areas only a few micrometers apart within each tail exposed to a given beam fluence. An analysis of the AgNOR variations in NBP and BP areas of tails exposed to different beam fluences revealed a dose-dependent effect. The AgNOR provide quantitative evidence of differential damage in neighboring tissue areas exposed to different LET regions of a helium-ion beam. PMID- 12541072 TI - A unifying model for reconstructing radiosensitivity from micronucleus formation, apoptosis and abnormal morphology. AB - At present micronucleus data cannot predict cellular radiosensitivity. The inclusion of data from apoptosis and abnormal morphology has not entirely resolved this problem. Here, we assess the probability of cell death arising from events other than micronucleation, apoptosis and abnormal morphology (i.e. lesions not detected by these damage assays) P(oe), for its ability to reflect intrinsic cellular radiosensitivity. Analysis of data from 17 cell lines used in two separate studies, spanning a wide range of radiosensitivity (0.0940 mm Hg). The SCTA depicted acute PE in all patients at T0 with complete resolution of endovascular clots in 10 patients at T1. At T0, the mean percentage of obstruction of the pulmonary arterial bed was significantly higher in the 22 patients with echocardiographic signs of severity (56+/-13 vs 28+/-32%; p<0.001). A significant reduction in the mean percentage of pulmonary artery obstruction was observed in the 19 patients with resolution of echocardiographic criteria of severity between T0 and T1 T0: 57+/-14%; T1: 7+/-11%; p<0.001). The threshold value for severe PE on CT angiograms was 49% (sensitivity: 0.773; specificity: 0.214). The mean (+/-SD) pulmonary artery pressure was significantly higher in the 26 patients with more than 50% obstruction of the pulmonary artery bed (45+/-15 mm Hg) than in the 10 patients with less than 50% obstruction of pulmonary artery bed at T0 (31+/-11 mm Hg; p<0.01). The CT severity score evaluated in the present study enables quantitative assessment of acute PE severity on spiral CT angiograms, readily applicable in routine clinical practice. PMID- 12541108 TI - Active disease and residual damage in treated Wegener's granulomatosis: an observational study using pulmonary high-resolution computed tomography. AB - The purpose of this study was to determine to what extent high-resolution computed tomography (HRCT) of the lungs can distinguish active inflammatory disease from inactive cicatricial disease in patients treated for Wegener's granulomatosis (WG). Twenty-eight WG patients with active pulmonary disease underwent a first HRCT examination immediately before standard immunosuppressive treatment and a second examination after clinical remission had been achieved. Lesions remaining after treatment were categorized as residual damage and were compared with findings during active disease to see by what features active and cicatricial disease can be distinguished. During active disease 17 patients had nodules/masses, 12 had ground-glass opacities, 6 had septal lines and 6 had non septal lines. After treatment, ground-glass opacities had resolved completely. Nodules/masses had resolved in 8 patients and had diminished in 7 patients. Residual nodules were distinguished from nodules/masses in active disease by lack of cavitation and a diameter of mostly <15 mm. In one-third of patients lines resolved, but in 8 instances new lines evolved during immunosuppression. During a follow-up period of a median 26.5 months (range 20.0-33.8), patients with residual nodules or lines had no more relapses than patients with completely cleared lungs. Treated pulmonary WG leaves substantial residual damage. High resolution CT does assist in the distinction between active and inactive lesions. Ground-glass opacities, cavitating nodules/masses and masses measuring more than 3 cm represent active disease ordinarily. Non-cavitary small nodules and septal or non-septal lines can be either active or cicatricial lesions. The nature of these lesions needs to be clarified by longitudinal observation. PMID- 12541109 TI - Thoracic manifestation of Wegener's granulomatosis: CT findings in 30 patients. AB - Our objective was to describe the CT findings of thoracic Wegener's granulomatosis. At presentation, both conventional and thin-section CT scans were available in 30 patients with Wegener's granulomatosis. Serial CT scans (range of intervals: 1-25 months, mean 4.5 months) were available in 20 patients. The initial and follow-up CT scans were analyzed retrospectively by two observers in terms of pattern and extent of parenchymal and airway lesions. Positive CT findings were seen in 29 of 30 (97%) patients at initial presentation. The most common pattern was nodules or masses seen in 27 of 30 (90%) patients. They were multiple in 23 of 27 (85%) patients, bilateral in 18 (67%), subpleural in 24 (89%), and peribronchovascular in 11 (41%) in distribution. Bronchial wall thickening in the segmental or subsegmental bronchi was seen in 22 (73%) patients. Large airways were also abnormal in 9 (30%) patients. Patchy areas of consolidation and ground-glass opacity were seen in 7 (23%) patients, respectively. In 17 of 20 (85%) patients in whom follow-up CT scans were available, the parenchymal or airway lesion showed complete or partial improvement with treatment. The CT findings of Wegener's granulomatosis, although multiple and variable, consist mainly of bilateral subpleural or peribronchovascular nodules or masses and bronchial wall thickening in the segmental or subsegmental bronchi. Parenchymal and airway lesions improve with treatment in most patients. PMID- 12541110 TI - Assessment of myocardial viability by MR imaging. AB - Diagnosis of myocardial viability after infarction focuses on the prediction of functional improvement of dysfunctional myocardium after revascularization therapy. Magnetic resonance imaging provides different approaches for the detection of myocardial viability. Measurement of end-diastolic wall thickness is easy to perform and has a high sensitivity, but a low specificity, and can only be used 4 months after myocardial infarction due to infarct healing processes. Low-dose dobutamine stress has a good sensitivity with a high specificity for the prediction of wall motion improvement, but this is only true for patients with a singular dysfunctional area and only slightly depressed cardiac function. Late enhancement allows for direct visualization of necrotic or scarred tissue. By measuring the transmural extent of late enhancement, the probability of mechanical improvement can precisely be given. Imaging of microvascular obstruction by first-pass perfusion or late enhancement gives additional information on viability and patient prognosis. Metabolic imaging techniques, such as (31)P-MR spectroscopy and (23)Na-MR imaging, provide further insights into the mechanisms of myocardial infarction and viability. In conclusion, cardiac MRI offers several clinically usable approaches for the assessment of myocardial viability and will probably become the method of choice in the near future. PMID- 12541111 TI - Relationships between coronary calcification detected at electron beam computed tomography and percutaneous transluminal coronary angioplasty results in coronary artery disease patients. AB - Our objective was to assess the relationship between coronary artery calcification and outcomes of percutaneous transluminal coronary angioplasty (PTCA). Electron beam computed tomography (EBCT) was performed in 80 patients with coronary artery disease (CAD) who underwent PTCA. The calcium score in each coronary artery vessel was estimated (in Agatston units) and compared with the occurrence of complications and restenosis. Angioplasty had been performed in 96 coronary artery segments with stenosis in 80 patients. The average calcium score in the unsuccessful PTCA segments was significantly higher than in ones with successful results. For complications these values were 65.0+/-79.9 and 27.0+/ 44.7, respectively ( p=0.02), and for restenosis 63.9+/-87.9 and 27.3+/-41.3, respectively ( p=0.03). Applying receiver operating characteristics analysis we determined sensitivity and specificity of EBCT for forecasting complications and restenosis. The best cut-off value of calcium score in segment of PTCA for prediction of restenosis was 27 (sensitivity 73%, specificity 67%), and for prediction of complications 29 (sensitivity 80%, specificity 74%). Coronary segments with extensive calcification were not optimal target lesions for PTCA. Analysis of coronary calcium score should be taken into consideration during planning of endovascular coronary interventions. PMID- 12541112 TI - Role of power Doppler techniques and ultrasound contrast enhancement in the differential diagnosis of focal breast lesions. AB - The purpose of this article is to demonstrate the diagnostic impact of ultrasound in differentiating focal breast lesions with special regard on power Doppler and US contrast agents. The sonographic evaluation of breast lesions has become a standard procedure during the past 15 years. Especially the improvement of B-mode resolution and the use of high-frequency probes increased the diagnostic value of US. Assuming that the neoangiogenetic vascular architecture of solid breast lesions can be depicted reliably by color Doppler, many authors tried to differentiate between benignity and malignancy using Doppler criteria such as flow and morphologic aspects. Additionally, adjuvant techniques, such as harmonic imaging and new US contrast agents, are meant to be success-promising tools. Whereas the sensitivity and specificity of color Doppler have varied in different studies, prognostic prediction and treatment monitoring seem to be the future areas of application. To evaluate sufficiently flow signals of very small vessels with low flow velocity, the use of contrast-enhancing agents may be necessary. Nevertheless, an indispensable condition for successful Doppler-based assessment of the entity of breast lesions is the standardization of techniques, evaluation, analysis and weighting of the parameters. PMID- 12541113 TI - Evaluation of neoadjuvant chemotherapeutic response of breast cancer using dynamic MRI with high temporal resolution. AB - The aim of this study was to evaluate changes in both size and contrast enhancement of breast tumors during neoadjuvant chemotherapy, using dynamic MRI with high temporal resolution. Patients with advanced breast cancer (n=21) underwent a 1.5-T MRI scan prior to and following neoadjuvant chemotherapy with four cycles. Dynamic contrast enhancement was measured using a fast turbo-FLASH sequence and quantified using a two-compartment model with the parameters k(ep) and amplitude. Image analysis was done on images overlayed with a color map of parameters. The correlation between tumor diameter measured by histopathology and MRI was 0.7 (p<0.003). A reduction of tumor size after chemotherapy of more than 25% was associated with a decrease of both analyzed contrast enhancement parameters (k(ep): p<0.002; amplitude: p<0.006), where k(ep) began to drop already after the first cycle of chemotherapy (p<0.008). A clear reduction of tumor size was only noted after the third cycle (p<0.008). In patients without tumor regression there was also a trend towards an early reduction of contrast enhancement. We assume that MRI with high temporal resolution and color mapping is a novel tool to assess therapeutic effects of neoadjuvant chemotherapy in breast tumors, which deserves further prospective evaluation. PMID- 12541114 TI - Phyllodes tumors of the breast. AB - Our objective was to study the color and pulse Doppler ultrasonography of phyllodes tumors of the breast. The data of 36 phyllodes tumors (19 benign, 5 borderline, and 12 malignant) were reviewed retrospectively. The median age of the patients was 41.5 years (range 13-62 years), and the tumors varied in size from 1.5 to 20 cm (median 4.6 cm). Most of the phyllodes tumors were lobulated masses with smooth margins, mildly hypoechoic internal echo texture, a heterogeneous internal echo pattern, and no microcalcification. Vessels were detectable on 35 of these tumors, with 51.4% having resistance index >0.700, 45.7% having pulsatility index >1.300, and 51.4% having V(max) >15 cm/s. The color Doppler characteristics were of no significant use in predicting the histological nature of the phyllodes tumors. If the sonographic features suggest a phyllodes tumor, a histological examination should be conducted to confirm the diagnosis. PMID- 12541115 TI - Galactography: method of choice in pathologic nipple discharge? AB - Our objective was to assess the role of galactography in the diagnostic work-up of patients with pathological nipple discharge together with exfoliative cytology, and investigation of secretion colour. All galactographies performed in this center between 1993 and 1998 were evaluated retrospectively. In 134 cases, outcomes, defined as either a postoperative histology or a negative follow-up over 2 years after galactography, were available. In 96 of these cases, exfoliative cytology was performed, and in 65 cases the colour of the secretion was determined. Galactograms, cytological findings and colour of the secretions were compared with the outcomes. At galactography, the radiographic findings had the following individual sensitivities/specificities in detecting carcinomas: filling defect 55.6%/62.1%; ductectasia 22.2%/94%; and filling stop 5.6%/77.6%. Normal galactograms (n=33, 25%) had a sensitivity of 78% and a specificity of 93% in predicting absence of disease. Technically inadequate investigations occurred in 16 (12%) cases. Cytology showed 69.2%/66.8% for papillomatous borderline or malignant cells; 53.8%/75.3% for borderline or malignant cells; and 7.7%/100% for one case of malignant cells. For the assessment of pathological secretions, galactography is a sensitive but unspecific method for the detection of papillomas or carcinomas. Filling defects, ductectasia and ductal distortion carry the highest levels of suspicion for carcinoma. A normal galactogram is a specific yet moderately sensitive indicator of absence of localized disease. Exfoliative cytology shows low sensitivity but better specificity for carcinomas when borderline or malignant cells are found. In the present series, the colour of the secretions does not add significant information about underlying pathology. PMID- 12541116 TI - Bowel MR imaging with oral Gastrografin: an experimental study with healthy volunteers. AB - Our objective was to evaluate Gastrografin for MR bowel imaging. Twenty-three healthy volunteers in two randomised groups received 300 or 400 ml 50% Gastrografin, drunk continuously during 2 and 3 h, respectively. Images were applied during breath-hold in three orthogonal orientations. The balanced fast field echo (BFFE) and balanced turbo field-echo (BTFE) sequences, with acquisition times from 13 to 25 s, were used before gadolinium (Gd) DTPA implying 1- to 2-mm-thick slices locally or 6-mm-thick slices through the entire gastrointestinal tract. The Gd-enhanced images were performed using a 3D T1 weighted FFE sequence with water selective excitation (Proset). Image quality, including bowel distention, homogeneity of opacification and wall conspicuity, were evaluated by two experienced reviewers, and the adverse reactions were recorded. Very good or excellent distention, homogeneity and wall conspicuity were achieved in the central segments from the ileum to the left colon flexure in 83-96% of cases, due to the adequate contrast media supply in these regions. Distention, homogeneity and delineation were good in the central segments of the remaining bowels. Diarrhoea was a major problem affecting all participants, followed by nausea. Provided that there is modern fast sequential technology, excellent MR imaging of the bowel can be achieved by the oral administration 50% diluted Gastrografin. Further studies are needed to refine the technique and optimise the quantity and concentration of Gastrografin in order to avoid or reduce adverse reactions. PMID- 12541117 TI - The impact of helical computed tomography on the diagnosis of unsuspected inflammatory bowel disease in the large bowel. AB - Inflammatory bowel diseases (IBD) are diagnoses that can be easily overlooked clinically. The aim of this study was to investigate if CT is able to make a contribution to the identification of previously unsuspected cases of IBD. We retrospectively identified cases in which the possibility of an IBD was raised in CT reports (over a 4-year period), by utilising a keyword search of the radiology database. Cases with a previously known or suspected IBD were rigorously excluded by review of case notes, and endoscopic, radiological, histological and microbiological findings. The CT images of the identified cases were reviewed by a blinded observer to document the extent of bowel wall thickening, the location of lesion(s), and presence of peri-colic fat abnormality, ascites and/or collections. The observer also attempted to corroborate the presence, and to identify the type, of IBD based on the CT appearances alone. Thirty-five cases (out of approximately 19,000 body CTs performed) of clinically unsuspected IBD were identified, of which 27 underwent further investigation. An IBD was confirmed in 48% (13 of 27): Crohn's disease (n=6), ulcerative colitis (n=2), pseudomembranous colitis (n=3) and other (n=2), of which 70% (9 of 13) were correctly typed by the reviewer. Inflammatory bowel disease was not substantiated in the remainder (14 of 27), although 7 of these had other bowel pathologies: diverticular disease (n=4); and carcinoma (n=3). Prospectively determining the presence, and furthermore type, of IBD on CT is challenging largely because of the considerable overlap in the appearances of the individual IBDs and indeed of normality. Nevertheless, CT is able to identify clinically unsuspected cases and radiologists should be alert to this treatable and not infrequently elusive diagnosis. PMID- 12541118 TI - Helical CT in acute lower gastrointestinal bleeding. AB - The purpose of this study was to assess the usefulness of helical CT in depicting the location of acute lower gastrointestinal bleeding. A three-phase helical CT of the abdomen was performed in 24 patients referred for acute lower gastrointestinal bleeding. The diagnosis of the bleeding site was established by CT when there was at least one of the following criteria: spontaneous hyperdensity of the peribowel fat; contrast enhancement of the bowel wall; vascular extravasation of the contrast medium; thickening of the bowel wall; polyp or tumor; or vascular dilation. Diverticula alone were not enough to locate the bleeding site. The results of CT were compared with the diagnosis obtained by colonoscopy, enteroscopy, or surgery. A definite diagnosis was made in 19 patients. The bleeding site was located in the small bowel in 5 patients and the colon in 14 patients. The CT correctly located 4 small bowel hemorrhages and 11 colonic hemorrhages. Diagnosis of the primary lesion responsible for the bleeding was made in 10 patients. Our results suggest that helical CT could be a good diagnostic tool in acute lower gastrointestinal bleeding to help the physician to diagnose the bleeding site. PMID- 12541119 TI - High signals in the uterine cervix on T2-weighted MRI sequences. AB - The aim of this pictorial review was to illustrate the normal cervix appearance on T2-weighted images, and give a review of common or less common disorders of the uterine cervix that appear as high signal intensity lesions on T2-weighted sequences. Numerous aetiologies dominated by cervical cancer are reviewed and discussed. This gamut is obviously incomplete; however, radiologists who perform MR women's imaging should perform T2-weighted sequences in the sagittal plane regardless of the indication for pelvic MR. Those sequences will diagnose some previously unknown cervical cancers as well as many other unknown cervical or uterine lesions. PMID- 12541120 TI - Ultrasound appearances of the testicular appendages: pictorial review. AB - Five testicular appendages are formed during development of the male genito urinary tract, which are the remnants of the degenerating mesonephric and paramesonephric ducts. The testicular and epididymal appendages, found at the upper pole of the testis and at the head of the epididymis respectively, are the most common and have a range of appearances on ultrasound. These appendages have the ability to undergo torsion, an important differential diagnosis in the child who presents with an acute scrotum. The varying ultrasound appearances of the testicular appendages are described and illustrated. Ultrasound features of appendiceal torsion are also demonstrated. PMID- 12541121 TI - Trans-catheter arterial chemoembolization as first-line treatment for hepatic metastases from endocrine tumors. AB - Our objective was to report the outcome in patients with liver metastasis from endocrine tumors who underwent transarterial chemoembolization (TACE) as first line non-surgical treatment. From January 1990 to December 2000, 14 patients with progressive unresectable liver metastases from digestive neuroendocrine tumor were treated with TACE (mean of 3.6 sessions) before any non-surgical treatment (somatostatin analogue, chemotherapy or interferon). Liver involvement was less than 50% in 11 patients. Size of the largest lesion ranged from 1.5 to 10 cm. Ten patients presented with carcinoid symptoms. The TACE was performed with Doxorubicin emulsified in Lipiodol and gelatin sponge particles. Symptomatic response upon flushes and/or diarrhea was complete in 7 of 10 cases and partial in 2 of 10 cases. An objective morphologic response was noted in 12 of 14 cases. The 5- and 10-year survival rate from diagnosis was 83 and 56%, respectively. Six patients were alive at the end of the study after 27-100 months from first TACE and 38-142 months from diagnosis. Three of them were successfully palliated for 55, 69, and 100 months with only TACE as treatment. Long-term palliation is possible in unresectable liver metastases from digestive neuroendocrine tumors with a few sessions of TACE as first-line and eventually exclusive treatment. PMID- 12541122 TI - Human fascioliasis: MR imaging findings of hepatic lesions. AB - Our objective was to describe MR imaging findings of liver lesions in human fascioliasis. The MR imaging of the liver was performed in 29 patients with fascioliasis. Seventeen patients were women and 12 were men, with a mean age of 47.5 years (age range 17-75 years). Hepatic lesions were grouped into five types based on their signal characteristics. Three patients had normal imaging findings. One or more lesions were observed in the other 26 patients. The lesion types and the frequency of appearances were as follows: hyperintensity of the liver capsule on T2-weighted images (n=16, 55.2%); ill-defined slightly hyperintense areas on T2-weighted images (n=18, 62.1%); lesions which were hypointense on T1-weighted and hyperintense on T2-weighted images (n=10, 34.5%); hypointense on T1-weighted images and centrally hypo- or hyperintense, surrounded by peripherally less hyperintense area on T2-weighted images (n=4, 13.8%); and hypointense foci or ill-defined hypointense areas on T1- and T2-weighted images (n=10, 34.5%). We describe the MR imaging features of the disease. Our findings may help the differential diagnosis in which fascioliasis should be added to the list. PMID- 12541123 TI - Pancreatic carcinoma: the role of high-resolution multislice spiral CT in the diagnosis and assessment of resectability. AB - The purpose of our study was to evaluate multislice computed tomography (MSCT) in the assessment of patients with clinical, laboratory, and US suspicion of pancreatic neoplasm, and to evaluate resectability status. Forty-six patients with a suspected pancreatic tumor underwent MSCT. After a preliminary precontrast survey, a postcontrast scan was performed in the arterial and portal venous phase with the following protocol: 4 x 1-mm collimation; 1.25- and 5-mm slice thickness width, respectively, and 1- and 5-mm reconstruction interval. In all patients pathological correlation was obtained. The evaluation of all images provided a diagnosis in 44 patients, with a sensitivity, specificity, and accuracy of 97, 80, and 96%, respectively. The MSCT correctly provided a diagnosis of unresectability with sensitivity of 96%, specificity of 86%, and accuracy of 93%. Evaluation of 1-mm slices demonstrated 83 of the 91 liver metastases found at surgery; conversely, the 5-mm slices detected only 76 of these lesions. Infiltration of peripancreatic major vessels was demonstrated, and was confirmed at surgery in 18 patients. High-resolution MSCT improves prediction of resectability in patients with suspected pancreatic carcinoma. Parenchymal and vascular information can be achieved with a single MSCT examination. PMID- 12541124 TI - Bone mineral density reference values in the normal female and male population of Izmir, Turkey. AB - In this study we aimed to establish the dual-energy X-ray absorptiometry (DXA) bone mineral density (BMD) reference values of healthy Turkish women and men residing in Izmir, western Turkey. We examined 347 women and 119 men (age range 20-80 years) who did not have any known risk factor that might affect bone mass. The BMD measurements were performed by Hologic QDR 4500 W Elite DXA instrument from the lumbar spine (L1-L4) and non-dominant hip. In women the mean BMD (g/cm(2))+/-standard deviation (SD) of the normal reference group between 20 and 39 years of age was 0.963+/-0.121 g/cm(2) in the lumbar spine and 0.891+/-0.119 g/cm(2) in total femoral region. The percentages of annual and total BMD losses in women between 30 and 80 years of age were 0.74 and 29% at spinal level and 0.65 and 26% in femoral region, respectively. The average BMD of the normal men's group between 20 and 39 years of age was 0.996+/-0.111 g/cm(2) in the lumbar spine and 1.025+/-0.110 g/cm(2) in femoral region. The percentages of annual and total BMD losses were 0.33 and 13% at spinal level and 0.50 and 20% in femoral region, respectively. In conclusion, like the values reported from mid-Anatolian region of Turkey, the BMD values of Turkish women and men residing in Izmir, western Anatolia, are also lower than in most European countries with regard to US and preinstalled Hologic values. PMID- 12541125 TI - Contrast-enhanced power Doppler ultrasonography of the metacarpophalangeal joints in rheumatoid arthritis. AB - The aim of this study was to examine, with dynamic contrast-enhanced MRI as the reference, if contrast-enhanced power Doppler ultrasonography (CE PDUS) of rheumatoid arthritis (RA) metacarpophalangeal (MCP) joints provides additional information for evaluation of synovial inflammation compared with PDUS. One MCP joint in each of 15 RA patients and 3 healthy control persons were examined with PDUS before and after intravenous bolus Levovist contrast injection. Corresponding rates of early synovial enhancement (RESE), previously shown to be closely related to histopathological synovitis, were calculated from dynamic contrast-enhanced MR images obtained the same day. Prior to ultrasonography, the joint was evaluated clinically. Levovist increased the flow signal in 7 of 9 joints with pre-contrast flow-signal and in 0 of 9 without pre-contrast signal. No healthy controls showed CE PDUS signal. The results of CE PDUS and dynamic MRI were closely related: RESE in joints with CE PDUS signal was significantly higher than in joints without CE PDUS signal (Mann-Whitney test, p<0.001). Among the patients with pre-contrast PDUS signal no statistically significant difference in RESE values was found between joints with and without post-contrast flow-signal increase. No correlation was found between clinical examination and CE PDUS. Based on comparisons with dynamic contrast-enhanced MRI, PDUS appears to be reliable for assessment of synovitis in RA MCP joints. Intravenous contrast injection may provide additional information in selected cases but did not in the present study increase the sensitivity of the method. PMID- 12541126 TI - Detection of postoperative residual cholesteatoma with delayed contrast-enhanced MR imaging: initial findings. AB - Our objective was to assess the value of delayed contrast-enhanced T1-weighted spin-echo MR imaging in the detection of residual cholesteatoma in patients who have undergone canal wall-up tympanoplasty procedure. The MR imaging was obtained prior to revision surgery in 18 patients with opacity of the post-operative cavity at CT examination 12-18 months after canal wall-up tympanoplasty. In each patient the following was performed: precontrast T1- and T2-weighted images; and early and delayed contrast-enhanced axial and coronal T1-weighted imaging. Early and delayed MR imaging results were separately compared with surgical second-look findings. Sensitivity, specificity, and predictive values were evaluated for early and delayed post-contrast MR imaging, compared with second-look surgery findings. A residual cholesteatoma was correctly identified in 8 of 9 cases with delayed contrast-enhanced T1-weighted MR imaging. Mean sensitivity, specificity, positive predictive value, and interobserver agreement (evaluated by kappa statistics) were, respectively, 85.2, 92.6, 92.6%, and kappa=0.78 for the delayed contrast-enhanced MR imaging technique. The same parameters were, respectively, 96.3, 33.3, 60.6, and 0.30 for the early contrast-enhanced T1-weighted MR images. We conclude that delayed contrast-enhanced T1-weighted MR imaging is reliable for the detection of residual cholesteatomas of the middle ear in patients who have undergone canal wall-up tympanoplasty. PMID- 12541127 TI - Usefulness of Doppler waveform analysis in differential diagnosis of cervical lymphadenopathy. AB - We compared Doppler spectral parameters in acute inflammatory, reactive, lymphomatous, and metastatic lymph nodes, and evaluated pulsed Doppler sonography as a method for distinguishing between different causes of cervical lymphadenopathy. Spectral Doppler analysis with measurements of resistance index (RI), pulsatility index (PI), peak systolic velocity (PSV), and end-diastolic velocity (EDV) was performed in 197 patients with cervical lymphadenopathy. Results of Doppler analysis were compared with findings of cytology and histology or with clinical presentation and follow-up. Student's t-test was used to assess statistical significance of differences in Doppler parameters between groups of patients. Significant differences for RI and PI were shown between all groups of patients except between lymphomatous and reactive nodes. Specificity of 100% for metastatic nodal involvement was shown for cutoff values RI>0.80 and PI>1.80. A positive predictive value (PPV) of 100% for acute lymphadenitis was shown for cutoff values RI<0.50 and PI<0.60. An EDV>9 cm/s has 100% negative predictive value for nodal metastasis, and EDV<1 cm/s has 100% specificity and PPV for metastasis. Although there exist differences in RI, PI, PSV, and EDV between different nodal diseases, none of these parameters offer both good sensitivity and good specificity, and only extreme cutoff values may occasionally be helpful in differential diagnosis. Doppler spectral analysis is a valuable noninvasive adjunct which can help in differentiation between metastatic, lymphomatous, acute inflammatory, and reactive lymphadenopathy, but cannot obviate biopsy in the majority of cases. PMID- 12541128 TI - Late adverse reactions to intravascular iodinated contrast media. AB - Late adverse reactions to intravascular iodinated contrast media are defined as reactions occurring 1 h to 1 week after contrast medium injection. They have received increasing interest over the past decade, but their prevalence remains uncertain and their pathophysiology is not fully understood. The Contrast Media Safety Committee of the European Society of Urogenital Radiology decided to review the literature and to issue guidelines. An extensive literature search was carried out and summarized in a report. Based on the available information, simple guidelines have been drawn up. The report and guidelines were discussed at the 8th European Symposium on Urogenital Radiology in Genoa. Late adverse reactions after intravascular iodinated contrast medium include symptoms such as nausea, vomiting, headache, itching, skin rash, musculoskeletal pain, and fever. A significant proportion of these reactions is unrelated to the contrast medium; however, allergy-like skin reactions are well-documented side effects of contrast media with an incidence of approximately 2%. Late reactions appear to be commoner after non-ionic dimers. The majority of late skin reactions after contrast medium exposure are probably T-cell-mediated allergic reactions. Patients at increased risk of late skin reactions are those with a history of previous contrast medium reaction and those on interleukin-2 treatment. Most skin reactions are self limiting and resolve within a week. Management is symptomatic and similar to the management of other drug-induced skin reactions. PMID- 12541129 TI - A prospective survey of delayed adverse reactions to iohexol in urography and computed tomography. AB - We investigated 7505 inpatients who underwent intravenous urography or contrast enhanced computed tomography to assess risk factors for delayed adverse drug reactions to iohexol, a non-ionic iodinated contrast medium. Focusing on delayed adverse reactions, all adverse events were prospectively investigated for 7 days after injection of iohexol. To explore the relevant risk factors, the relationship between occurrence of adverse reactions to iohexol and 17 different variables was evaluated by logistic regression analysis. To assess the influence of seasonal factors, adverse reactions were separately evaluated during two periods: February to April (the pollinosis period in Japan) and July to September (the non-pollinosis period). The prevalence of delayed adverse events and delayed adverse reactions was 3.5 and 2.8%, respectively, whereas the prevalence of adverse events and adverse reactions was 5.7 and 5.0%, respectively. Multivariate analysis showed that six parameters had a significant influence on delayed adverse reactions to iohexol, including (a) a history of allergy, (b) season, (c) radiographic procedure, (d) age, (e) concomitant surgery or other invasive procedures, and (f) concomitant medication. The prevalence of delayed reactions was lower than in previous large-scale studies. Significant risk factors included a history of allergy and performance of radiography during the pollinosis period, suggesting that allergy was involved in delayed adverse reactions. The type of radiographic procedure also had an influence. PMID- 12541130 TI - Fluorescence imaging with near-infrared light: new technological advances that enable in vivo molecular imaging. AB - A recent development in biomedical imaging is the non-invasive mapping of molecular events in intact tissues using fluorescence. Underpinning to this development is the discovery of bio-compatible, specific fluorescent probes and proteins and the development of highly sensitive imaging technologies for in vivo fluorescent detection. Of particular interest are fluorochromes that emit in the near infrared (NIR), a spectral window, whereas hemoglobin and water absorb minimally so as to allow photons to penetrate for several centimetres in tissue. In this review article we concentrate on optical imaging technologies used for non-invasive imaging of the distribution of such probes. We illuminate the advantages and limitations of simple photographic methods and turn our attention to fluorescence-mediated molecular tomography (FMT), a technique that can three dimensionally image gene expression by resolving fluorescence activation in deep tissues. We describe theoretical specifics, and we provide insight into its in vivo capacity and the sensitivity achieved. Finally, we discuss its clinical feasibility. PMID- 12541131 TI - Percutaneous injection of human thrombin to treat iatrogenic femoral pseudoaneurysms: short- and midterm ultrasound follow-up. AB - Our objective was to determine the efficacy of ultrasound-guided direct injection of human thrombin to close postcatheterization pseudoaneurysms and to evaluate the midterm results of this new percutaneous technique by ultrasound follow-up. One hundred one postcatheterization pseudoaneurysms in 100 patients were treated by direct injection of human thrombin (mean dose 200 IU, range 50-750 IU) under ultrasound guidance. Patient demographics, clinical data and ultrasonographic morphology of the pseudoaneurysms were analyzed. Midterm (mean 99 days, range 25 210 days) ultrasonographic follow-up was performed in 57 of 82 called patients (69.5%). Ninety-six of 101 pseudoaneurysms thrombosed completely after the first procedure. Of the remaining 5 pseudoaneurysms, 3 were treated successfully after the second attempt, and 2 patients underwent surgical repair; thus, the primary success rate was 95% (96 of 101), and the overall success rate was 98% (99 of 101). No changes in distal pulses or other adverse events were noted immediately or 1 day after the procedure. Ultrasound follow-up revealed complete disappearance of the pseudoaneurysm in 70% (40 of 57), presence of small, residual groin hematoma in 26% (15 of 57), and partial reperfusion of the pseudoaneurysm in 3.5% (2 of 57). Human thrombin injection under ultrasound guidance is a very effective and safe method of therapy. Failure (immediate and late) of the technique as well as procedure-related complications are very rare. In our institution, sonographically guided human thrombin injection has become the treatment of choice for postcatheterization pseudoaneurysms. PMID- 12541132 TI - Interaction of normal saline solution with ultrasound contrast medium: significant implication for sonographic diagnosis of vesicoureteral reflux. AB - The sonographic diagnosis of vesicoureteral reflux with intravesical administration of ultrasound contrast media is on the rise. In few cases did we encounter rapid dissolution of microbubbles in the bladder filled with normal saline. The aim of this study was to find out whether there is any interaction between normal saline and ultrasound contrast media and, if so, to elucidate the cause for the interaction. In an in vitro experimental setup the mixtures of various normal saline solutions and a galactose-based ultrasound contrast medium (Levovist) were scanned under pre-defined parameters. The duration of contrast was determined. Oxygen concentration of the solutions was measured (pO(2) and O(2) mg/l). The US contrast medium had significantly longer contrast duration when mixed with normal saline from plastic containers rather than glass containers. The contrast duration difference was even more between normal saline from containers sealed under vacuum and those not sealed under vacuum: 0.7 and 12 min, respectively. The oxygen concentration in normal saline from vacuum-sealed containers is one-third or less that from non-vacuum-sealed containers. The high concentration of dissolved oxygen in the normal saline seems to prevent the diffusion of air from the microbubbles into the solution and thus their collapse. Desaturated normal saline has detrimental effect on microbubbles of the galactose based US contrast medium. For the purpose of filling the bladder during contrast enhanced voiding urosonography only normal saline solution from non-vacuum-sealed containers should be used. For practical purposes, normal saline from plastic containers is safest. PMID- 12541134 TI - Tumour-specific activation of the sodium/iodide symporter gene under control of the glucose transporter gene 1 promoter (GTI-1.3). AB - Targeted transfer of a functionally active sodium iodide symporter (NIS) into tumour cells may be used for radioiodine therapy of cancer. Therefore, we investigated radioiodine uptake in a hepatoma cell line in vitro and in vivo after transfer of the sodium iodide symporter ( hNIS) gene under the control of a tumour-specific regulatory element, the promoter of the glucose transporter 1 gene (GTI-1.3). Employing a self-inactivating bicistronic retroviral vector for the transfer of the hNIS and the hygromycin resistance genes, rat Morris hepatoma (MH3924A) cells were infected with retroviral particles and hNIS-expressing cell lines were generated by hygromycin selection. (125)I(-) uptake and efflux were determined in genetically modified and wild type hepatoma cells. In addition, the iodide distribution in rats bearing wild type and genetically modified hepatomas was monitored. hNIS-expressing MH3924A cell lines accumulated up to 30 times more iodide than wild type hepatoma cells, with a maximal iodide uptake after 30 min incubation time. Competition experiments in the presence of sodium perchlorate revealed a decrease in the iodide uptake (80-84% decrease). Moreover, ouabain led to a loss of accumulated I(-) (81% decrease) whereas 4,4'-diisothiocyano-2,2' disulphonic acid stilbene (DIDS) increased the I(-) uptake into cells (87% increase). However, a rapid efflux of the radioactivity (70%) was observed 20 min after (125)I(-)-containing medium had been replaced by non-radioactive medium. Lithium had no significant effect on iodide efflux. In rats, the hNIS-expressing tumours accumulated 22 times more iodide than the contralateral wild type tumour. In accordance with the in vitro data, we also observed a rapid efflux of the radioactivity out of the tumour in vivo. Dosimetric calculations resulted in an absorbed dose of 85 mGy in the wild type tumour and 830 mGy in the hNIS expressing tumour after administration of 18.5 MBq (131)I. In conclusion, transduction of the hNIS gene under the control of the GLUT1 promoter element induces iodide transport in Morris hepatoma cells in vitro and in vivo. However, for therapeutic application additional conditions need to be defined which inhibit the iodide efflux out of the tumour cells. PMID- 12541136 TI - Comparison of methods for thyroid volume estimation in patients with Graves' disease. AB - Individualised dosage models are frequently applied for radioiodine therapy in patients with Graves' hyperthyroidism, especially in Europe. In these dosage schemes the thyroid volume is an important parameter. Thyroid volume determinations are usually made with ultrasonography or with thyroid scintigraphy, although the accuracy of planar scintigraphy for this purpose is not well established. The aim of this study was to compare the accuracy of three modalities for the determination of the thyroid volume in patients with Graves' disease: planar scintigraphy (PS), single-photon emission tomography (SPET) and ultrasonography (US). These three modalities were compared with magnetic resonance imaging (MRI) as the gold standard. Thyroid volume estimations were performed in 25 patients with Graves' disease. The PS images were subjected to filtering and thresholding, and a standard surface formula was used to calculate the thyroid volume. With SPET the iteratively reconstructed thyroid images were filtered, and after applying a threshold method an automatic segmentation algorithm was used for the volume determinations. Thyroid volumes were estimated from the US images using the ellipsoid volume model for multiple two-dimensional measurements. For MRI, thyroid segmentation was performed manually in gadolinium enhanced T1-weighted images and a summation-of-areas technique was used for the volume measurements. The thyroid volumes calculated with MRI were 25.0+/-13.8 ml (mean+/-SD, range 7.0-56.3 ml). PS correlated poorly with MRI ( R(2)=0.61) and suffered from a considerable bias (-4.0+/-17.6 ml). The differences between PS and MRI volume estimations had a very large spread (33+/-58%). For SPET both the correlation with MRI ( R(2)=0.84) and the bias (1.8+/-11.9 ml) were better than for PS. US had by far the best correlation with MRI ( R(2)=0.97) and the best precision, but the bias (6.8+/-7.5 ml) was not negligible. In conclusion, SPET is preferred over PS for accurate measurements of thyroid volume. US is the most accurate of the three modalities, if a correction is made for bias. PMID- 12541135 TI - Defining the success of cardiac gene therapy: how can nuclear imaging contribute? AB - Gene therapy is a promising modality for the treatment of various cardiovascular diseases such as ischaemia, heart failure, restenosis after revascularisation, hypertension and hyperlipidaemia. An increasing number of approaches are moving from experimental and preclinical validation to clinical application, and several multi-centre trials are currently underway. Despite the rapid progress in cardiac gene therapy, many basic tools and principles remain under development. Questions with regard to the optimal method for gene delivery in a given situation remain open, as do questions concerning therapeutic efficacy and the time course and magnitude of gene expression in target and remote areas. Nuclear imaging provides valuable tools to address these open issues non-invasively. Functional effects of molecular therapy at the tissue level can be identified using tracers of blood flow, metabolism, innervation or cell death. The use of reporter genes and radiolabelled reporter probes allows for non-invasive assessment of location, magnitude and persistence of transgene expression in the heart and the whole body. Co-expression of a reporter gene will allow for indirect imaging of the expression of a therapeutic gene of choice, and linkage of measures of transgene expression to downstream functional effects will enhance the understanding of basic mechanisms of cardiac gene therapy. Hence, nuclear imaging offers great potential to facilitate and refine the determination of therapeutic effects in preclinical and clinical cardiovascular gene therapy. PMID- 12541137 TI - Nuclear medicine technologist training in European countries. PMID- 12541138 TI - Reflex connections from forearm and hand afferents to shoulder girdle muscles in humans. AB - Using surface electromyographic recordings from the trapezius and serratus anterior muscles and percutaneous electrical stimulation of the median, ulnar and radial nerves, the reflex connections from forearm and hand afferents to these shoulder girdle muscles have been investigated in normal human subjects. Stimulation of the median, ulnar and radial nerves at the shoulder, elbow and wrist evoked late, excitatory reflexes in the upper and lower parts of trapezius and in serratus anterior. These reflexes are not evoked by stimulation of cutaneous afferents alone, since there was no response to stimulation of the distal cutaneous branches of these three nerves. Measurements of the conduction velocity of afferents of the median, ulnar and radial nerve evoking these reflexes gave a mean conduction velocity of approximately 50 m/s. The lowest stimulus intensities at which these reflexes could be evoked were found to be 0.3 times motor threshold (MT). By taking into account the reflex latency, the length of the conduction path and that the reflex was mediated by low threshold, fast conducting afferents, it is proposed that group I muscle afferents from the forearm or hand evoke a supraspinal reflex to trapezius and serratus anterior. It appears that the functional significance of these reflexes is to aid in the stability of the shoulder girdle. PMID- 12541139 TI - Object-based representations facilitate memory for inhibitory processes. AB - Previous work has shown that in a sequential cueing task, inhibition of the return of attention (IOR) can be observed for up to four or five loci. We have argued that the inhibition processes mediating IOR are associated with object based representations, and it is object-based representations that are maintained in memory. Experiments presented here show that, compared with standard conditions in which a number of identical grey squares are cued, cueing empty locations tends to reduce the memory for prior inhibitory processes; while cueing objects which are distinctive in colour and shape tends to increase memory for inhibition. Converging with other recent findings, we conclude that memory for the inhibitory processes of attention facilitates visual search and that this memory is dependent on object-based representations. PMID- 12541140 TI - Innocuous cooling can produce nociceptive sensations that are inhibited during dynamic mechanical contact. AB - In a previous study of the heat grill illusion, sensations of burning and stinging were sometimes reported when the skin was cooled by as little as 2 degrees C. Informal tests subsequently indicated that these nociceptive sensations were experienced if cooling occurred when the stimulating thermode rested on the skin, but not when the thermode was cooled and then touched to the skin. In experiment 1 subjects judged the intensity of thermal (cold/warm) and nociceptive (burning/stinging) sensations when the volar surface of the forearm was cooled to 25 degrees C (1) via a static thermode (Static condition), or (2) via a cold thermode touched to the skin (Dynamic condition). The total area of stimulation was varied from 2.6 to 10.4 cm(2) to determine if the occurrence of nociceptive sensations depended upon stimulus size. Burning/stinging was rated 10.3 times stronger in the Static condition than in the Dynamic condition, and this difference did not vary significantly with stimulus size. In experiment 2, thermal and nociceptive sensations were measured during cooling to just 31 degrees, 29 degrees or 27 degrees C, and data were obtained on the frequency at which different sensation qualities were experienced. Stinging was the most frequently reported nociceptive quality in the Static condition, and stinging and burning were both markedly reduced in the Dynamic condition. In experiment 3 we tested the possibility that dynamic contact might have inhibited burning and stinging not because of mechanical contact per se, but rather because dynamic contact caused higher rates of cooling. However, varying cooling rate over a tenfold range (-0.5 degrees to -5.0 degrees /s) had no appreciable effect on the frequency of stinging and burning. Overall, the data show that mild cooling can produce nociceptive sensations that are suppressed under conditions of dynamic mechanical contact. The latter observation suggests that cold is perceived differently during active contact with objects than during passive heat loss to the environment. Hypotheses about the physiological basis of the nociceptive sensations at mild temperatures and their possible role in the phenomena of paradoxical heat and synthetic heat are discussed. PMID- 12541141 TI - Inhibition of saccade initiation by preceding smooth pursuit. AB - In this study, we investigated the influence of smooth-pursuit eye movements on saccade initiation in response to a sudden jump of a continuously moving target. We replicated the finding by Tanaka et al. (1998) that saccadic eye movements in the direction opposite to preceding pursuit have longer latencies than those in the same direction. We confirmed that this asymmetry is indeed due to an inhibitory effect of smooth pursuit on saccade initiation in the opposite direction rather than facilitation of saccade initiation in the same direction. The inhibitory effect decreased strongly when subjects knew the jump direction in advance. This supports the notion that the prolonged latencies of backward saccades are not due to orbital mechanics or low-level motor processing. Furthermore, we found that the range of saccade directions inhibited by a pursuit movement is broad, covering all directions that did not have the same horizontal component as the pursuit direction. This is in contrast with the predictions of "Inhibition of Saccade Return" (ISR, Hooge and Frens 2000), which is restricted to a smaller confined area. PMID- 12541142 TI - Proprioceptive sensory function in Parkinson's disease and Huntington's disease: evidence from proprioception-related EEG potentials. AB - In both Parkinson's disease and Huntington's disease, proprioceptive sensory deficits have been suggested to contribute to the motor manifestations of the disease. Here, proprioceptive sensory function was investigated in Parkinson's disease patients, Huntington's disease patients, and healthy control subjects (each group n=8), using proprioception-related evoked potentials. Proprioception related potentials were elicited by passive index finger movements and measured with high-density EEG. Conventional median nerve somatosensory evoked potentials (mnSEPs) were recorded in the same session. Analysis included amplitude and latency measures from selected scalp electrodes and dipole source reconstruction. We found a proprioception-related N90 component of normal latency in both Parkinson's disease and Huntington's disease. The source strength of the underlying cortical generator was normal in Parkinson's disease, but marginally reduced in Huntington's disease. Using the source location of the N20-P20 component of the mnSEP as a landmark for postcentral area 3b, the N90 was localized to the precentral motor cortex. At a latency around 170-180 ms proprioception-related potentials were explained by bilateral sensory cortex activation with an altered distribution in Parkinson's disease and a reduction of ipsilateral activation in Huntington's disease. Together, the results show largely normal early proprioception-related potentials, but changes in the cortical processing of kinaesthetic signals at longer latencies in both diseases. PMID- 12541143 TI - Comparison between the effects of chronic and acute hypoxemia on muscle afferent activities from the tibialis anterior muscle. AB - The reflex loops initiated by the activation of muscle afferents are altered by a reduction of the oxygen supply. This has been shown in different mammalian species under experimental conditions of acute or chronic hypoxemia. In the present study in rats, we compared the effects of acute and chronic hypoxemia on the activity of afferents from the tibialis anterior muscle to investigate the existence of possible adaptive mechanisms to hypoxaemia. The activity in groups I and II (mechanosensitive) and groups III and IV (mechano- and mostly chemosensitive) muscle afferents was recorded under conditions of normoxaemia and acute and chronic hypoxemia. Chronic hypoxaemia was achieved by exposing the rats for 45 consecutive days to a gas mixture containing 10% oxygen in nitrogen, whereas acute hypoxaemia was limited to a 1-h exposure to a 10% oxygen mixture. Different test agents: muscle stimulation (MS), arterial injection of KCl or lactic acid and tendon vibrations, were used to activate muscle afferents. Both acute and chronic hypoxaemia suppressed the post-MS activation of groups III and IV muscle afferents and significantly depressed the response of these afferents to KCl and lactic acid. The pattern of response of mechanosensitive afferents to mechanical tendon vibration was similar under the three conditions but chronic hypoxemia significantly reduced the response to 10-50 Hz vibrations. We conclude that inhibitory effects on muscle metabosensitive afferents occurred in the first minutes following acute hypoxemia and was prolonged with chronic hypoxemia. Our data also suggest that chronic hypoxemia depresses mechanosensitive muscle afferents. This may explain the observation that sensorimotor control of skeletal muscles is often markedly impaired in hypoxemic humans. PMID- 12541144 TI - Visual-tactile spatial interaction in saccade generation. AB - Saccadic reaction times to visual targets tend to be faster when non-visual stimuli are presented in close temporal or spatial proximity even if subjects are instructed to ignore the accessory input. The effect tends to decrease with increasing spatial distance between the stimuli. Multisensory interaction effects measured in neural structures involved in saccade generation have demonstrated a similar spatial dependence. The present study investigated visual-tactile interaction effects on saccadic reaction time using a focused attention paradigm. Compared to unimodal visual targets saccadic reaction time to bimodal stimuli was reduced by up to 30 ms. The effect was larger for ipsi- than for contralateral presentations, and it increased with the eccentricity of the visual target. The results are consistent with attributing part of the facilitation to a multisensory effect of bimodal neurons with overlapping visual and tactile receptive field structures in the deep layers of the superior colliculus. PMID- 12541145 TI - Effects of predictive mechanisms on head stability during forward trunk perturbation. AB - While much is known about reflex and mechanical contributions to the control of head stability, little is known about predictive control. The goal of this experiment was to determine the contribution of predictive mechanisms to head stability in space, in the pitch plane, during forward trunk perturbations. Eleven standing healthy subjects had their trunk pulled forward by a load-pulley apparatus. The perturbation was either self-triggered or imposed (triggered by the experimenter). Subjects were exposed to two loads: 2% and 4% of their body weight. The contributions of torques acting on the head-neck system were inferred from head and trunk kinematics, neck muscle EMG, and the torques acting on the head, which were computed using inverse dynamics. The results showed that both the head and trunk moved less during the self-triggered than imposed condition during both loads for most of the participants. There was no evidence of predictive neck countertorque or increased neck muscle co-contraction during the self-triggered condition. These findings suggest that most of the subjects improved head stability in the self-triggered condition by reducing trunk motion and the associated interactive torque that perturbed the head. PMID- 12541146 TI - Object motion perception is shaped by the motor control mechanism of ocular pursuit. AB - It is still a matter of debate whether the control of smooth pursuit eye movements involves an internal drive signal from object motion perception. We measured human target velocity and target position perceptions and compared them with the presumed pursuit control mechanism (model simulations). We presented normal subjects (Ns) and vestibular loss patients (Ps) with visual target motion in space. Concurrently, a visual background was presented, which was kept stationary or was moved with or against the target (five combinations). The motion stimuli consisted of smoothed ramp displacements with different dominant frequencies and peak velocities (0.05, 0.2, 0.8 Hz; 0.2-25.6 degrees /s). Subjects always pursued the target with their eyes. In a first experiment they gave verbal magnitude estimates of perceived target velocity in space and of self motion in space. The target velocity estimates of both Ns and Ps tended to saturate at 0.8 Hz and with peak velocities >3 degrees /s. Below these ranges the velocity estimates showed a pronounced modulation in relation to the relative target-to-background motion ('background effect'; for example, 'background with' motion decreased and 'against'-motion increased perceived target velocity). Pronounced only in Ps and not in Ns, there was an additional modulation in relation to the relative head-to-background motion, which co-varied with an illusion of self-motion in space (circular vection, CV) in Ps. In a second experiment, subjects performed retrospective reproduction of perceived target start and end positions with the same stimuli. Perceived end position was essentially veridical in both Ns and Ps (apart from a small constant offset). Reproduced start position showed an almost negligible background effect in Ns. In contrast, it showed a pronounced modulation in Ps, which again was related to CV. The results were compared with simulations of a model that we have recently presented for velocity control of eye pursuit. We found that the main features of target velocity perception (in terms of dynamics and modulation by background) closely correspond to those of the internal drive signal for target pursuit, compatible with the notion of a common source of both the perception and the drive signal. In contrast, the eye pursuit movement is almost free of the background effect. As an explanation, we postulate that the target-to-background component in the target pursuit drive signal largely neutralises the background to-eye retinal slip signal (optokinetic reflex signal) that feeds into the eye premotor mechanism as a competitor of the target retinal slip signal. An extension of the model allowed us to simulate also the findings of the target position perception. It is assumed to be represented in a perceptual channel that is distinct from the velocity perception, building on an efference copy of the essentially accurate eye position. We hold that other visuomotor behaviour, such as target reaching with the hand, builds mainly on this target position percept and therefore is not contaminated by the background effect in the velocity percept. Generally, the coincidence of an erroneous velocity percept and an almost perfect eye pursuit movement during background motion is discussed as an instructive example of an action-perception dissociation. This dissociation cannot be taken to indicate that the two functions are internally represented in separate brain control systems, but rather reflects the intimate coupling between both functions. PMID- 12541147 TI - Upregulation of FGF-2 in reactive spinal cord astrocytes following unilateral lumbar spinal nerve ligation. AB - Spinal nerve ligation results in dramatic changes in spinal cord primary C afferent fibers, which include atrophy with an accompanied decrease in calcitonin gene-related peptide (CGRP). These changes parallel the activation of astrocytes, which have been implicated in the ensuing neuropathic pain states. As part of an effort to elucidate the role of the downstream effectors of astrocyte reactivity in the context of allodynia, the expression of fibroblast growth factor-2 (FGF-2) was examined following tight ligation of L5 and L6 spinal nerves. FGF-2 is a pleiotropic cytokine that is synthesized and secreted by neurons and astrocytes. FGF-2 immunoreactivity was increased in ipsilateral dorsal horn reactive astrocytes at 1 and 3 weeks following nerve ligation. Semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) of laser-captured dorsal spinal cord sections revealed an increase in FGF-2 mRNA in the dorsal horn ipsilateral to nerve injury compared to contralateral and SHAM. Furthermore, an increase in FGF-2 mRNA in ispilateral dorsal root ganglia (DRG) was seen by in situ hybridization. These results demonstrate that, in response to ligation-induced injury of sensory neurons, FGF-2 is upregulated in both DRG neurons and in spinal cord astrocytes, suggesting neurotrophic functions of this growth factor following peripheral nerve lesion and possibly in astrocyte-related maintenance of pain states. PMID- 12541148 TI - Effect of movement speed on limb segment motions for reaching from a standing position. AB - The performance of a standing reaching task that necessitates some forward bending requires: (1) the coordination of multiple joints (i.e., the trunk and limb segments) to reach the target, and (2) the preservation of postural stability. It has been proposed that the neural control of multijoint reaching tasks can be simplified by time scaling of joint motions while keeping joint excursions the same. To determine if time scaling of joint motions was used in this more complex reaching task, we had 20 healthy subjects (10 male and 10 female) reach for two targets located in a parasagittal plane while standing on a force platform. Subjects reached for the targets at a comfortable speed and a fast paced speed. Sagittal plane motions of the right shank, thigh, pelvis, trunk, humerus, and forearm were measured. At the fast paced movement speeds subjects had significantly larger excursions of the thigh, pelvis, humerus, and forearm compared to the comfortable speed. Thus, segment motions are not simply time scaled for standing multijoint reaches. We explored three possible reasons for not obeying time scaling: (1) to reduce scaling of peak kinetic energy, (2) to reduce scaling of peak horizontal ground reaction force, and (3) a convergence of movement strategies at faster speeds. While subjects modified their movement strategy in relationship to movement speed, these changes had no significant effect on the expected scaling of peak kinetic energy, or peak horizontal ground reaction forces. Given the intersubject differences in movement strategies used to perform these reaching tasks at the fast speeds, a convergence of movement strategies was ruled out. We propose that the increase in segment motions with speed may be a consequence of rules underlying motor output, the increases being greater for segments in which viscoelastic resistance to movement is more significant compared to inertial resistance. PMID- 12541149 TI - Features of motor performance that drive adaptation in rapid hand movements. AB - In order to explore how subjects correct for errors in movement and adapt their motor programs, we studied rapid hand movements. Subjects grasped a grooved knob and made brisk turning movements to various targets, similar to tuning a radio dial. A motor attached to the knob shaft was configured to apply a destabilizing negative viscous perturbation. Following a period of practice with no perturbations, the negative viscosity was engaged, which caused a large change in overall kinematics: the peak velocity increased, the movement amplitude was too large, and discrete corrective submovements were generated to bring the pointer back onto the target. After about an hour and nearly 1000 trials, subjects learned to move accurately in the new dynamic environment, returning their overall kinematics near to previous levels. Measures of performance included the endpoint error of the primary movement (the initial movement segment), the frequency and amplitude of corrective submovements, task success rate, mean squared jerk, and deviation from a "normal" angular velocity temporal profile. Both the amplitude and frequency of corrective submovements decreased progressively during adaptation as the subjects made fewer target overshoot errors. These results are consistent with motor learning schemes in which adaptation of the motor controller is driven by an attempt to reduce the endpoint error of the primary movement. While there have been many theories regarding what is being optimized in motor control, in general, biologically plausible mechanisms for implementing these schemes have not been described. A biologically plausible optimization criterion is the minimization of the occurrence and amplitude of corrective submovements, since the latter have been proposed as realistic climbing fiber training signals for adaptive changes in the cerebellum. We postulate that the other criteria that have been proposed are instead secondary to an increased accuracy of the primary movement and a corresponding decrease in the occurrence and amplitude of corrective submovements. PMID- 12541150 TI - Intersegmental synchronization of spontaneous activity of dorsal horn neurons in the cat spinal cord. AB - Extracellular recordings of neuronal activity made in the lumbosacral spinal segments of the anesthetized cat have disclosed the existence of a set of neurons in Rexed's laminae III-VI that discharged in a highly synchronized manner during the occurrence of spontaneous negative cord dorsum potentials (nCDPs) and responded to stimulation of low-threshold cutaneous fibers (<1.5x T) with mono- and polysynaptic latencies. The cross-correlation between the spontaneous discharges of pairs of synchronic neurons was highest when they were close to each other, and decreased with increasing longitudinal separation. Simultaneous recordings of nCDPs from several segments in preparations with the peripheral nerves intact have disclosed the existence of synchronized spontaneous nCDPs in segments S1-L4. These potentials lasted between 25 and 70 ms and were usually larger in segments L7-L5, where they attained amplitudes between 50 and 150 micro V. The transection of the intact ipsilateral hindlimb cutaneous and muscle nerves, or the section of the dorsal columns between the L5 and L6, or between the L6 and L7 segments in preparations with already transected nerves, had very small effects on the intersegmental synchronization of the spontaneous nCDPs and on the power spectra of the cord dorsum potentials recorded in the lumbosacral enlargement. In contrast, sectioning the ipsilateral dorsal horn and the dorsolateral funiculus at these segmental levels strongly decoupled the spontaneous nCDPs generated rostrally from those generated caudally to the lesion and reduced the magnitude of the power spectra throughout the whole frequency range. These results indicate that the lumbosacral intersegmental synchronization between the spontaneous nCDPs does not require sensory inputs and is most likely mediated by intra- and intersegmental connections. It is suggested that the occurrence of spontaneous synchronized nCDPs is due to the activation of tightly coupled arrays of neurons, each comprising one or several spinal segments. This system of neurons could be involved in the modulation of the information transmitted by cutaneous and muscle afferents to functionally related, but rostrocaudally distributed spinal interneurons and motoneurons, as well as in the selection of sensory inputs during the execution of voluntary movements or during locomotion. PMID- 12541151 TI - Short-latency eye movements evoked by near-threshold galvanic vestibular stimulation. AB - To investigate whether the primary planes of eye and body responses to galvanic vestibular stimulation (GVS) are congruent, we have measured the binocular, three dimensional eye movements (scleral coil technique) to bilateral bipolar GVS in six normal human subjects. Stimulation intensities were kept deliberately low in order to characterize the response to near-threshold intensities of stimulation (0.1-0.9 mA) that had been used previously to characterise body postural responses. Stimuli were applied for 4 s, but only the early responses that occurred within the initial 300 ms of turning the current on or off were measured. At intensities of 0.1-0.7 mA the 'on' response consisted almost exclusively of a torsional slow phase eye movement in which the top of the eyes rotated towards the anode. The latency of the torsional response was ca. 46 ms. A weak polarity-dependent disconjugate response was also observed in which the intorting eye elevated and the extorting eye depressed ('skew eye deviation'). When the current was turned off similar responses occurred in the reverse direction. Removal of the visual fixation light-emitting diode (LED) had no consistent effect on the short-latency ocular responses. The direction of the ocular response was similar to that of the postural response and is compatible with GVS inducing an apparent dynamic roll-tilt of the head towards the cathode. However, weak horizontal eye movements, which became more prominent as the stimulus intensity was increased to 0.9 mA, were also observed. This suggests that an additional weak rotational component about the yaw axis, or a component of lateral translation in the frontal plane, is contained in the GVS-evoked signal. The overall pattern of eye movement suggests that semicircular canal afferents contribute to these low-intensity GVS responses. PMID- 12541152 TI - Comparison of cardiac output measured with echocardiographic volumes and aortic Doppler methods during mechanical ventilation. AB - OBJECTIVE: To compare two transesophageal echocardiographic methods of cardiac output and stroke volume measurement in mechanically ventilated patients. DESIGN: Prospective clinical study. SETTING: Operating room (group I) and intensive care unit (group II) in two university hospitals. PATIENTS: Fifteen deeply anesthetized patients undergoing gynecological laparoscopy for sterility (group I) and 40 patients with septic shock (group II). INTERVENTIONS: Transesophageal echocardiography with modification of hemodynamic conditions. MEASUREMENTS AND RESULTS: Left ventricular (LV) volumes, cardiac (CI) and stroke index (SI) were measured with two methods using either LV volumes or aortic Doppler. These values were significantly lower in group I compared to group II. Using ANOVA and paired t-tests, there were no significant differences between the two methods of measurement. Correlation between these methods was better in group II than in group I, although not significantly so. In group I, bias for CI measurements was low (0.05 l/min per m(2)), with a weak agreement in terms of the 95% confidence interval (-1.17; 1.06 l/min per m(2)) compared to the mean values obtained with both methods (1.3 l/min per m(2)). In group II, bias for CI measurements was lower (0.2 l/min per m(2)). Agreement was weak, regarding 95% confidence intervals (-1.7; 1.3 l/min per m(2)) compared to the mean values (3 l/min per m(2) with the LV volumes method and 3.2 l/min per m(2) and with the Doppler method). CONCLUSIONS: Cardiac output and stroke volume can be measured from LV volumes in mechanically ventilated patients, yielding relevant information. However, the accuracy of LV volume measurements is not excellent compared to the aortic Doppler method. Thus, this latter technique should still be considered as the gold standard. PMID- 12541153 TI - The ANZPIC registry diagnostic codes: a system for coding reasons for admitting children to intensive care. AB - OBJECTIVE: To describe the uniform diagnostic coding system used in Australia and New Zealand to code reasons for admitting children to intensive care, and to highlight the benefits of a uniform approach. DESIGN: International, multicentre, observational study. SETTING: A registry of children admitted to intensive care in Australia and New Zealand. PATIENTS: The records of 19249 children admitted to intensive care between 1997 and 2000 were analysed. MEASUREMENTS AND RESULTS: The system was designed empirically using expert consensus. The principal diagnosis or main reason for intensive care admission and up to five associated diagnoses are coded. The system has four levels of coding: non-operative or post-procedural admission, diagnostic group, specific condition, and for injury and infection the aetiological factor. The main reason for intensive care admission was coded in all patient records, however, for 11.1% of records the code was limited to diagnostic group with the specific condition coded as "other diagnosis". Two or more diagnoses were coded in 61% of records. The most frequent reason for admission was asthma. CONCLUSIONS: The major advantage of the system is that units in the region use the same method of coding. A uniform international approach to coding reasons for admitting children to intensive care is needed. PMID- 12541154 TI - PIM2: a revised version of the Paediatric Index of Mortality. AB - OBJECTIVE: To revise the Paediatric Index of Mortality (PIM) to adjust for improvement in the outcome of paediatric intensive care. DESIGN: International, multi-centre, prospective, observational study. SETTING: Twelve specialist paediatric intensive care units and two combined adult and paediatric units in Australia, New Zealand and the United Kingdom. PATIENTS: All children admitted during the study period. In the analysis, 20787 patient admissions of children less than 16 years were included after 220 patients transferred to other ICUs and one patient still in ICU had been excluded. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: A revised model was developed by forward and backward logistic regression. Variable selection was based on the effect of including or dropping variables on discrimination and fit. The addition of three variables, all derived from the main reason for ICU admission, improved the fit across diagnostic groups. Data from seven units were used to derive a learning model that was tested using data from seven other units. The model fitted the test data well (deciles of risk goodness-of-fit chi(2 )8.14, p=0.42) and discriminated between death and survival well [area under the receiver operating characteristic (ROC) plot 0.90 (0.89-0.92)]. The final PIM2 model, derived from the entire sample of 19638 survivors and 1104 children who died, also fitted and discriminated well [chi(2 )11.56, p=0.17; area 0.90 (0.89-0.91)]. CONCLUSIONS: PIM2 has been re calibrated to reflect the improvement that has occurred in intensive care outcome. PIM2 estimates mortality risk from data readily available at the time of ICU admission and is therefore suitable for continuous monitoring of the quality of paediatric intensive care. PMID- 12541155 TI - Effects of positive end-expiratory pressure increments can be predicted by computer simulation based on a physiological profile in acute respiratory failure. AB - OBJECTIVE: We examined whether computer simulation predicts airway pressures after increments of positive end-expiratory pressure (PEEP) in acute respiratory failure. DESIGN AND SETTING: Prospective, nonrandomized comparative trial in an intensive care unit of a university hospital. PATIENTS: Twelve consecutive acute respiratory failure patients. INTERVENTIONS. PEEP increments from 0 to 2.5, 5, 7.5, 10, and 15 cm H(2)O. MEASUREMENTS AND RESULTS: A physiological profile comprising values for compliance, respiratory resistance and CO(2 )elimination as a function of tidal volume was established from a recording of ordinary breaths prior to increments of PEEP. Airway pressures and CO(2 )elimination were measured 30 s after resetting, pressures also after 10 min. Values from simulation of the resetting, based on the profile, were compared to measured values. The profiles indicated vast differences in physiology between the 12 subjects. Errors of simulation of airway pressures were nonsignificant or trivial up to PEEP levels of 10 cm H(2)O (95% of errors <3 cm H(2)O). After 10 min plateau pressure averaged 1.5 cm H(2)O lower than 30 s after resetting. At increments to PEEP 7.5, 10, and 15, CO(2 )elimination fell by on average 4%, 8%, and 11%, respectively. As tidal volume and respiratory rate was unchanged this was not predicted. CONCLUSIONS: On the basis of a simple lung model, simulation predicted effects of moderate increments of PEEP on airway pressures in patients with complex physiology. PMID- 12541156 TI - DCL-Hb for trauma patients with severe hemorrhagic shock: the European "On-Scene" multicenter study. AB - OBJECTIVE: A major cause of death in patients with severe hemorrhagic shock following trauma is the subsequent occurrence of multiple organ failure due to tissue hypoxia. Early administration of an oxygen carrier may reduce the occurrence of organ failures and improve survival. It may also reduce the need of blood products. DESIGN AND SETTING: Prospective multicenter study in a university clinic. PATIENTS: 121 patients with severe hemorrhagic shock. INTERVENTIONS: Patients were randomly assigned "on-scene" to receive either up to 1000 ml of a 10% diaspirin cross-linked hemoglobin (DCLHb) solution or the study center's standard therapy. MEASUREMENTS AND RESULTS: Demographic and physiological characteristics of the two treatment groups at baseline were comparable. Organ failures and survival rates until day 5 and day 28 showed no significant differences. The sponsor therefore terminated this trial prematurely after an interim evaluation of the data indicated no evidence of efficacy to offset concerns raised about the safety of DCLHb. Median volumes of cumulative blood products administered on 1 (1595 vs. 3716 ml) and 7 days (3139 vs. 4746 ml) after admission were lower in the DCLHb group. CONCLUSIONS: The early application of an oxygen carrier (DCLHb) to patients with severe hemorrhagic shock following trauma had no significant effect on the occurrence of organ failure or on 5- and 28-day survival in this abbreviated trial. However, early infusion of up to 1000 ml DCLHb reduces the need for blood products without changing morbidity or survival. PMID- 12541157 TI - Bactericidal activity response of blood neutrophils from critically ill patients to in vitro granulocyte colony-stimulating factor stimulation. AB - OBJECTIVE: Neutrophil function impairment is common in nonneutropenic critically ill patients. Whether granulocyte colony-stimulating factor (G-CSF) may be useful for preventing nosocomial infection in these patients is debated. The response of blood neutrophils from critically ill patients to G-CSF was investigated in vitro. DESIGN AND SETTING: Prospective study, laboratory investigation in two intensive care units. PATIENTS: 52 critically ill patients without immunosuppression. MEASUREMENTS: Neutrophils obtained from 52 patients on the 5th day of their intensive care unit stay were incubated with and without G-CSF (1, 10, 100 ng/ml). Reactive oxygen species (ROS) release and bactericidal activity against Staphylococcus aureus and Pseudomonas aeruginosa were evaluated. Plasma cytokines (interleukin 10, tumor necrosis factor alpha, and G-CSF) were measured. RESULTS: Median values (25th-75th percentiles) indicated no stimulatory effect of G-CSF on neutrophil bactericidal activity against either organism: S. aureus, 100% (95-109) of the unstimulated condition with 1 ng/ml G-CSF, and P. aeruginosa, 102% (98-109) with 1 ng/ml G-CSF. However, wide interindividual variability was found, ranging from marked inhibition to marked stimulation. Similar variability was found for ROS release. No correlations were found between ROS release and bactericidal activities against either bacterial strain. Inhibition of neutrophil bactericidal activity by G-CSF was associated with significantly higher plasma interleukin 10 concentrations. Plasma G-CSF levels were significantly higher in patients whose neutrophil bactericidal activity was unresponsive to G-CSF, suggesting G-CSF receptor downregulation. CONCLUSIONS: The effect of G-CSF on in vitro neutrophil bactericidal activity varied widely, depending on endogenous levels of G-CSF and was not predictable based on severity scores. PMID- 12541159 TI - Irreversible ischemia of the hand after peripheral administration of tromethamol (THAM). PMID- 12541158 TI - Monitoring of cerebral perfusion pressure during intracranial hypertension: a sufficient parameter of adequate cerebral perfusion and oxygenation? AB - OBJECTIVE: A cerebral perfusion pressure (CPP) oriented treatment is a widely accepted standard for patients with intracranial hypertension. In an animal model of controlled intracranial hypertension we investigated whether CPP is a reliable parameter of sufficient cerebral perfusion and oxygenation. Using near-infrared reflexion spectroscopy the effect of decreasing CPP due to increasing intracranial pressure (ICP) on cerebral tissue oxygenation was studied. METHODS: Ten rabbits were subjected to artificially elevated ICP using the cisterna-magna infusion technique. Regional cerebral O(2) saturation of hemoglobin (tiSO(2)), regional tissue concentration of hemoglobin (tiHb), and CPP were recorded continuously. CPP was investigated with respect to tiSO(2). Electrocortical activity was simultaneously recorded by two-channel EEG to determine the onset of ischemia. RESULTS: Reduced CPP due to increased ICP led to a continuous decrease in tiSO(2.) There was progressive suppression of EEG frequency and amplitude with decreasing CPP in all animals. Onset of EEG-silence due to elevated ICP was observed in a wide range of CPP-values between 9 and 42 mmHg. At the same time tiSO(2) varied merely between 0 and 5%. CONCLUSIONS: Regarding the EEG effects due to increased ICP (EEG silence), CPP values showed a wide interindividual variability, in contrast to tiSO(2). In our animal model the sole calculation of CPP did not reflect adequate cerebral perfusion. PMID- 12541160 TI - Base deficit in immediate postoperative period of coronary surgery with cardiopulmonary bypass and length of stay in intensive care unit. AB - OBJECTIVE: To assess the relationship between the base deficit value in the immediate postoperative period of coronary surgery for cardiopulmonary bypass and the length of stay in the ICU. DESIGN AND SETTING: Prospective descriptive study in the department of anesthesia and cardiovascular surgery of a university hospital. PATIENTS: 185 consecutive patients. INTERVENTIONS: Coronary artery bypass graft with cardiopulmonary by pass. MEASUREMENTS AND RESULTS: Thirty variables were determined during the pre-, intra-, and postoperative periods; a statistical univariate analysis was performed differentiating patients whose length of stay in the ICU was 2 days or less and those whose stay was more than 2 days. Secondly, a logistic regression model was performed on the variables shown to have a statistically significant difference in univariate analysis, with determination of the odd ratio. Fourteen variables had a statistically significant difference in univariate analysis and three of them highlighted by the logistic regression model: administration of catecholamines, base deficit value in the 1st h postoperatively, and age with odd ratios, respectively, of 3.15, 1.51, and 1.07). CONCLUSIONS: The value of base deficit measured during the 1st h after coronary surgery for cardiopulmonary bypass is correlated with the length of stay in ICU. PMID- 12541161 TI - Minocycline hypersensitivity reaction with acute respiratory distress syndrome. PMID- 12541162 TI - Clinical significance of pulmonary artery occlusion pressure. AB - BACKGROUND: Ppao values are routinely used to assess pulmonary vascular status and LV performance. Regrettably, under many common clinically relevant conditions, even when Ppao values are measured accurately, Ppao values at baseline and in response to therapy often reflect an inaccurate measure of cardiovascular status. RESULTS AND CONCLUSIONS: Thus, caution should be used when applying measures of Ppao in determining therapy if changes in RV volume, hyperinflation, or LV diastolic compliance are simultaneously occurring. PMID- 12541166 TI - [Anti-TNF antibodies in the treatment of inflammatory intestinal stenoses in Crohn's disease]. AB - Stenoses are a frequent complication in patients with Crohn's disease and represent a major diagnostic and therapeutic challenge. The proper assessment of the nature of a stenosis as inflammatory or fibrotic is critical for appropriate treatment, since symptomatic fibrotic stenoses require surgical resection. Standard diagnostic procedures to assess the nature of a stenosis include endoscopy, conventional contrast radiography and magnetic resonance tomography. Recent data suggest, that the positron-emission-tomography possesses a high sensitivity and specificity to confirm inflammatory activity in the bowel. The recombinant monoclonal anti-TNF-antibody Infliximab (Remicade) has been approved for the treatment of steroid refractory and steroid dependent Crohn's disease in Germany since 9/2000 and the efficacy of Infliximab is well documented. However, few data exist about the treatment of inflammatory stenoses with Infliximab. We performed a retrospective analysis of our experience with Infliximab in patients with Crohn's disease with special reference to patients with inflammatory stenoses. Among a total of 21 patients treated with Infliximab 11 patients had an inflammatory stenosis. 9 of these patients responded well to Inflimab and became completely asymptomatic for a considerable period of time. Infliximab was tolerated well except for one patient who developed an intrabdominal abscess. The notable clinical response of patients with inflammatory stenoses to Infliximab suggests that treatment with Infliximab might be helpful to postpone or avoid surgical intervention. This finding should be further investigated in a prospective randomized study. PMID- 12541165 TI - [Ceftriaxone versus Levofloxacin for antibiotic therapy in patients with acute cholangitis]. AB - INTRODUCTION: For the therapy of acute cholangitis complete biliary drainage and antibiotic therapy is needed. The aim of the current study was to compare intravenous therapy of acute cholangitis with Ceftriaxone or Levofloxacin in a prospective and randomized fashion. METHODS: Patients with biliary obstruction and clinical signs of infection received in addition to 1.5 g Metronidazole either 500 mg Levofloxacin/die or 2 g Ceftriaxone/die. Early on during ERCP, bile was aspirated via the cannulation catheter and cultured for bacteria under aerobic and anaerobic conditions. Minimal inhibitory concentrations of the respective antibiotics were determinate for each isolate. The clinical course was followed for at least 6 days with clinical and laboratory data. RESULTS: 60 patients with clinical signs of acute cholangitis were randomised. In 40 patients (66 %) biliary colonization with bacteria could be identified. In all bacterial species Levofloxacin showed significantly lower rates of in-vitro resistance as compared to Ceftriaxone. However, the percentage of patients with a clinical cure or significant improvement was the same in the two groups. CONCLUSIONS: The clinical effect of Levofloxacin and Ceftriaxone in patients with acute cholangitis showed no significant differences. Because of improved in-vitro efficiency, a calculated therapy with Levofloxacin might be advantageous. PMID- 12541167 TI - [Diagnostics and treatment of Crohn's disease -- results of an evidence-based consensus conference of the German Society for Digestive and Metabolic Diseases]. PMID- 12541168 TI - [Clinical diagnostics, classification]. PMID- 12541169 TI - [Pathologic diagnostics]. PMID- 12541170 TI - [Acute episode]. PMID- 12541171 TI - [Chronically active Crohn's disease]. PMID- 12541172 TI - [Maintaining a remission]. PMID- 12541173 TI - [Surgery -- fistulas]. PMID- 12541174 TI - [Psychosomatics]. PMID- 12541175 TI - [Complementary therapies]. PMID- 12541176 TI - [Extraintestinal manifestations]. PMID- 12541177 TI - [Nutrition]. PMID- 12541178 TI - [4199 biopsies from the endoscopic normal lower duodenum]. AB - Lower duodenal biopsies (LDB) are not taken at every oesophago-gastro duodenoscopy (EGD). In the present study, biopsies from the endoscopic normal lower duodenum were checked as a measure of quality assurance. From 1996 to 2000, 9,955 EGD were performed and 4,199 LDB were taken (42.2 %). Of these, 667 showed pathological histology (15.9 %). A non-specific inflammation was seen in 537 cases and lymphangiectasia in 30 cases. Signs of indigenous sprue were described histologically in 6 LDB. In 4 of the 6 first diagnoses, the LDB was taken owing to clinical suspicion of malabsorption syndrome. Giardia lamblia could be detected in 22 patients. Only 6 of the 22 patients had diarrhoea. A total of 18 clinically relevant first diagnoses were made by LDB in asymptomatic patients with normal endoscopic findings in the duodenum. In order to make a relevant first diagnosis, 233 LDB had to be taken. LDB can be dispensed within EGD when there is neither diarrhoea nor loss of weight, and no anemia, iron deficiency, vitamin deficiency, macrocytosis, hypoproteinaemia, meteorism, joint symptoms or fever. PMID- 12541179 TI - [Live demonstrations of endoscopic examinations -- a status determination]. PMID- 12541180 TI - [NASH -- nonalcoholic steatohepatitis]. AB - Nonalcoholic steatohepatitis describes a hepatic disorder with the typical characteristics of an alcoholic pathogenesis without alcohol consumption. It was first described in 1962 and named NASH by Ludwig et al. 1980. Many researchers worked on this disease since this time. It represents the hepatic manifestation of the syndrome X. The pathogenesis is a two-hit phenomenon. The first hit leads to steatosis hepatis and makes the liver vulnerable to the second hit. Central factors of the second hit are oxygen-radicals, oxidative stress, lipid peroxidation and cytokines. The exact pathogenic mechanisms are still unknown. NASH is a hepatic disease which can end up in liver cirrhosis and liver failure. Up to now a curative drug therapy does not exist. The poor prognosis in some cases, the increasing incidence in western populations and the lack of therapeutic options renders NASH to a serious problem. The aim of this article is to show the actual knowledge of this disease, especially focussed on the pathogenesis, by review of the literature from 1979 up to the present time. PMID- 12541184 TI - Albright's hereditary osteodystrophy and pseudohypoparathyroidism. AB - Pseudohypoparathyroidism (PHP) and Albright's hereditary osteodystrophy (AHO) are not interchangeable terms. AHO describes a constellation of physical features, including short adult stature, obesity, brachydactyly, and ectopic ossifications. PHP means end-organ resistance to PTH and is subclassified into types Ia, Ib, and Ic and type II. Pseudopseudohypoparathyroidism (PPHP) is a term used for individuals with AHO who have normal end-organ responses to PTH. Both the PHPIa and PPHP forms of AHO result from heterozygous deactivating mutations in the GNAS1 gene associated with a 50% reduction in bioactivity of the Gsalpha protein that it encodes. The GNAS1 gene is subject to tissue-specific genomic imprinting. Patients with mutations on their maternally derived allele are likely to have associated PHPIa, whereas mutations on the paternal allele usually cause PPHP. Isolated PTH resistance (PHPIb) can result from mutations within the GNAS1 gene but is more commonly caused by epigenetic imprinting abnormalities affecting the upstream exon 1A. The causes of PHP type Ic and PHP type II are not yet clear, and the latter is likely to be heterogeneous. PMID- 12541185 TI - Drug-induced metabolic bone disorders. AB - A number of drugs, either physician prescribed or those taken without the knowledge of the physician, may have metabolic effects on, and result in abnormal changes in, the musculoskeletal system. These medications may involve the developing fetus, infant, child, or adult and may manifest as drug-induced embryopathies, osteoporosis, osteomalacia, or rickets, or a combination of these, or they may be associated with proliferative changes. Not uncommonly, the radiologist may be the first physician to identify the metabolic manifestations of these medications. The radiological changes associated with such drugs will be reviewed. Certain drugs may have teratogenic effects, nonteratogenic effects, or both. These effects, dependent solely on the timing of administration, will be discussed separately. PMID- 12541186 TI - Osteopetrosis. AB - Osteopetrosis is a rare sclerosing inherited dysplasia of bone caused by the deficient function of osteoclasts. At first the disease was divided into the severe infantile recessive and the more benign autosomal dominant types, but clinical differences and progress in genetic understanding have now enabled identification of two distinct autosomal dominant types. The eponym Albers Schonberg disease should not be used as a generic term for the disease, because the single case reported almost 100 years ago almost certainly had an autosomal dominant inheritance. The diagnosis remains radiographic, supported by computed tomography (CT), if necessary. The use of magnetic resonance imaging (MRI) tends to be limited to imaging of the marrow in the severe recessive disease, which is usually fatal without marrow transplantation. PMID- 12541187 TI - Idiopathic hyperphosphatasia. AB - Idiopathic hyperphosphatasia is a rare autosomal recessive bone disorder, characterized by excessive bone resorption and bone formation. The radiographic appearances include widening of the diaphyses, vertebral osteoporosis, acetabular protrusion, and thickening of the skull vault. There is considerable variability in phenotype, with some cases diagnosed in infancy and others in later childhood. Most cases appear to arise from inactivating mutations in the gene encoding osteoprotegerin, a product of osteoblasts that is critically involved in osteoclastogenesis. Treatment with inhibitors of bone resorption (calcitonin or bisphosphonates) is successful in ameliorating some aspects of the disorder. PMID- 12541188 TI - Paget's disease of bone. AB - Paget's disease of bone is one of the most common diseases to affect bone, yet it is rare before the age of 50. The etiology of the condition remains unproven. Paget's disease of bone has become less common, less severe, and less extensive in recent decades. Isotope bone scans and radiographs remain the most frequent radiological investigations, demonstrating the extent of the disease and characteristic appearances in most cases. Recent changes in the radiological investigation of Paget's disease include increasing use of computed tomography (CT) and magnetic resonance (MR) imaging for the evaluation of less typical disease or disease complications; the incidental finding of Paget's disease on CT or MR requires recognition to avoid inappropriate investigation. The presence of sclerotic Paget's disease in the lumbar spine or hip may elevate bone mineral density measurements at these sites, with consequent potential to underestimate fracture risk. Awareness of the normal level-to-level vertebral variation in bone density in the spine, and careful assessment of the images acquired on dual energy X ray absorptiometry or quantitative CT will help to avoid this pitfall. Examples of these investigations and the combination of Paget's disease with other conditions such as osteoarthritis, metastatic bone disease, and bone infection are illustrated. PMID- 12541189 TI - Nuclear medicine studies in metabolic bone disease. AB - The principal application of nuclear medicine in metabolic bone disease is the isotope bone scan. Often, it is not a diagnostic tool but can be useful in clarifying the nature of a clinical problem. The best-established role for the bone scan in metabolic bone disease is in Paget's disease, in which it is diagnostic, provides definition of the extent of disease, and probably reflects disease activity. The isotope bone scan is also important in osteoporosis, for which it is not diagnostic but may often provide useful information to confirm that fracture has occurred, determine the age of the fracture, identify unsuspected fractures, and identify other causes for pain, for example, facet joint disease. The bone scan is less useful in other metabolic bone diseases, for example, renal osteodystrophy and osteomalacia, but will often have a characteristic appearance, with several metabolic features. The degree of positivity of the scan generally relates to the severity of the hyperparathyroidism. In patients with metabolic bone disease, other specific clinical problems may arise, such as osteomyelitis or avascular necrosis, and the bone scan may be diagnostic in these conditions. Nuclear medicine techniques are also of value in hyperparathyroidism and to localize the site of the adenoma where surgical treatment is being considered; the use of technetium Tc 99m sestamibi is now routine. More recently, positron emission tomography (PET) scanning has been found to be helpful in the more difficult cases. PMID- 12541190 TI - Tumoral calcinosis: 12 years later. AB - During the last 12 years, tumoral calcinosis (TC) has become an entity that is better understood by radiologists, clinicians, and pathologists. Several aspects of TC are well documented by the scientific community, although the most important, etiology, remains an enigma. As a consequence, the treatment of TC continues to be symptomatic. Imaging continues to be the best method for diagnosis, assessing extension, evaluating complications, monitoring treatment, and selecting patients for surgical excision. Investigators from different countries are reporting the most elusive component of TC: calcific myelitis. New systemic and cerebrovascular complications, even in the young, have been described recently. Similarly, ocular manifestation, which had not been reported previously, is well described in the recent literature. A few, poorly documented reports are confounding TC myelitis with osteomyelitis and TC periarticular soft tissue masses with tumoral calcium pyrophosphate dihydrate (CPPD) of the temporomandibular joint and spine. Advances in the clinical arena have resulted in a new approach and classification of several clinical forms of TC. Most of the recent literature has been related to the histopathology of TC. A new view of the old "traumatic theory" suggests microtrauma of connective tissues around the joints, causing bleeding and the formation of rudimentary bursae as the initial event. Perhaps, in the near future, research will define at the genetic and molecular levels the etiology and pathophysiology that will allow the treatment and prevention of TC. PMID- 12541191 TI - Metabolic disease in animals. AB - Rickets is a metabolic bone disorder characterized by osteopenic changes resulting from the failure of calcification of the osteoid matrix and absent mineralization of hypertrophic cartilage cells at the epiphyseal growth plates in growing primates, herbivores, swine, carnivores, and birds. The causes of rickets include inadequate dietary provision of calcium, phosphorus, and vitamin D. Osteomalacia in reptiles, simian bone disease in nonhuman primates, and osteodystrophia fibrosa (secondary hyperparathyroidism) or "bran disease" in herbivores are caused by a diet that has a much higher content of phosphorus than calcium, combined with inadequate exposure to direct sunlight. Medullary bone consists of interconnected spicules of bone resembling embryonic bone and is established in relation to the shell formation cycle of laying birds. Hypertrophic osteodystrophy develops in large-breed growing dogs, chickens, and guinea pigs and is possibly caused by vitamin C deficiency. Tibial dyschondroplasia is a defect in endochondral ossification characterized by a widened proximal tibial physis that is not penetrated by metaphyseal vascular sprouts, commonly found in growing broiler chickens, turkeys, and exotic birds. PMID- 12541193 TI - Imaging in metabolic bone disease. PMID- 12541194 TI - Vitamin D metabolism, rickets, and osteomalacia. AB - Rickets in the growing child or adolescent and osteomalacia in the adult develop in a variety of clinical situations and have in common an absence or delay in the mineralization of growth cartilage and in newly formed bone collagen. Classically, deficiency of vitamin D, which is essential for the absorption of dietary calcium, has been the major cause. However, rickets is also seen as a result of hereditary defects in critical vitamin D signaling molecules. Disturbances of phosphate metabolism can also lead to signs of rickets and osteomalacia, notably X-linked hypophosphatemic rickets, and oncogenic osteomalacia. Extrarenal synthesis of 1,25-dihydroxyvitamin D, such as that associated with granulomatous disease, can also lead to disturbances in calcium metabolism, with associated skeletal and nonskeletal changes. PMID- 12541195 TI - Rickets in childhood. AB - Bone accretion is a two-stage process, with the osteoblasts laying down osteoid, which is then mineralized. Mineralization of osteoid requires vitamin D to be available in its active form (1,25(OH) 2 D); in addition, mineralization also requires normal levels of serum calcium, phosphorus, and alkaline phosphate. Deficiencies of any of these will result in defective mineralization of bone, in which the mineral-osteoid ratio is reduced (a qualitative abnormality). In the juvenile skeleton deficiencies particularly affect enchondral ossification at the growth plates, giving the characteristic clinical and radiological features of rickets. The bone may be soft, leading to deformity. In this article the causes of rickets in childhood are reviewed, together with the clinical and radiological features and strategies for prevention and treatment. PMID- 12541196 TI - Oncogenic osteomalacia. AB - Oncogenic osteomalacia is a rare paraneoplastic syndrome caused by bone and soft tissue tumors. The characteristic clinical, metabolic, and imaging findings are reviewed, as is the radiologist's role in evaluation and diagnosis. New insight into the pathophysiology of oncogenic osteomalacia is also presented. PMID- 12541197 TI - Radiology of osteoporosis. AB - Osteoporosis is a common metabolic bone disorder that may be broadly subdivided into generalized or regional osteoporosis. This article reviews the common causes of generalized osteoporosis with emphasis on the radiological appearances, complications, and differential diagnosis. The more important causes of regional osteoporosis such as disuse osteoporosis, reflex sympathetic dystrophy, and transient regional osteoporosis are also discussed, and their imaging features are reviewed. PMID- 12541198 TI - Dual energy x-ray absorptiometry and its clinical applications. AB - Bone density scans using the technique of dual energy X-ray absorptiometry (DXA) are widely seen as having an essential role in the evaluation of patients at risk of osteoporosis. DXA scans of the spine and hip are the most useful because fractures at these sites result in the greatest impairment of quality of life. A hip DXA scan is the most reliable way of evaluating hip fracture risk, and the spine is the most sensitive site for monitoring response to treatment. Fundamental to the clinical role of DXA scanning are the findings of epidemiological studies that relate fracture risk to bone mineral density. The results of spine and hip DXA scans are interpreted using the World Health Organisation (WHO) definition of osteoporosis as a T-score less than -2.5. DXA scans are also used in longitudinal studies, both for research studies and for follow-up scans of individual patients. Although the former have an important role in the development of new treatments for osteoporosis, the latter are more controversial and are likely to be less important in the future. There is also interest in the use of smaller, less expensive DXA systems for scanning sites in the peripheral skeleton. However, in general, the results from these devices cannot be interpreted using the WHO definition of osteoporosis, and until there is a consensus over establishing an equivalent threshold, their use is premature. PMID- 12541199 TI - Quantitative computed tomography. AB - Quantitative computed tomography (QCT) is an established technique for measuring bone mineral density (BMD) in the axial spine and peripheral skeleton (forearm, tibia). QCT can determine in three dimensions the true volumetric density (mg/cm 3) of trabecular or cortical bone at any skeletal site. However, because of the high responsiveness of spinal trabecular bone and its importance for vertebral strength, QCT has been principally employed to determine trabecular BMD in the vertebral body. QCT has been used for assessment of vertebral fracture risk, measurement of age-related bone loss, and follow-up of osteoporosis and other metabolic bone diseases. This article reviews the current capabilities of QCT at different skeletal sites and the recent technical developments, including volumetric acquisition. PMID- 12541200 TI - Quantitative ultrasound in osteoporosis. AB - Quantitative ultrasound (QUS) has been applied to the assessment of bone for almost two decades. The physical interaction of ultrasound and bone is complex and not completely understood. However, it is considered that QUS provides information on bone mass and structure. Although dual-energy X-ray absorptiometry (DXA) is currently considered the "gold standard" for the diagnosis of osteoporosis, through the World Health Organisation definition, there is, in some parts of the world, limited availability to axial DXA. This has stimulated interest in smaller, more portable, and less expensive techniques to indentify those at risk of osteoporotic fracture. One such quantitative method is QUS, which has the added advantage over other photon absorptiometric bone density techniques of not using ionizing radiation. This article reviews the capabilities of QUS and indicates its appropriate application in clinical practice, based on current review of the scientific literature. PMID- 12541201 TI - Bone densitometry in children. AB - Compared with adults, limited attention has been paid to bone densitometry in children. However, due to the recognition of the importance of peak bone mass and the increased capabilities to treat children with diseases that affect bone growth and/or metabolism, bone densitometry is used more often in children. This article will discuss the available bone densitometry techniques and the limitations of their use in children. The article is concluded with a discussion on specific problems that can be encountered in pediatric bone densitometry. PMID- 12541202 TI - Identification of vertebral fractures in osteoporosis. AB - Osteoporosis is a systemic skeletal disease characterized by reduced bone mass and microarchitectural deterioration. These lead to subsequent increase in bone fragility and susceptibility to low-trauma or atraumatic fractures, most commonly vertebral fractures but also fractures of hip and wrist. These have a significant impact on morbidity, mortality, and health care cost. Studies have demonstrated that the presence of vertebral fractures is an independent and significant predictor of the increased risk for further fractures. The occurrence of a vertebral fracture is often clinically asymptomatic, and many of these fractures, therefore, remain undiagnosed. Recently, a number of techniques have been developed that allow for reliable identification of vertebral fractures on radiographs. The two most widely used methods in clinical research are the semiquantitative assessment of vertebral deformities, which is based on visual evaluation, and the quantitative approach, which is based on different morphometric criteria. In our practice for osteoporosis evaluation we use the Genant semiquantitative approach-an accurate and reproducible method, tested and applied in many clinical studies. The newest generation of fan-beam dual energy X ray absorptiometry (DXA) systems delivering "high-resolution" lateral spine images offer a potential practical alternative to radiographs for clinical vertebral fracture analysis. The advantages of using DXA over conventional X-ray devices are its minimal radiation exposure and high-speed image acquisition. It also allows combined evaluation of vertebral fracture status and bone mineral density, which could become a standard for patient assessment in osteoporosis. PMID- 12541203 TI - Imaging of trabecular bone structure. AB - In addition to bone mass, trabecular bone architecture is an important entity in assessing bone fragility, which is crucial in the diagnosis of osteoporosis. A number of imaging techniques have been used to analyze bone structure noninvasively. Projection radiography has been used with good results in the peripheral skeleton; however, this is only a two-dimensional technique, which reflects trabecular bone structure to a certain extent. High-resolution tomographic techniques, such as high-resolution magnetic resonance imaging and computed tomography (CT), have a limited spatial resolution but the potential to image three-dimensional architecture of trabecular bone. With the advances in magnetic resonance hardware and software and new CT techniques (i.e., multislice spiral CT and clinical micro-CT), noninvasive imaging of trabecular bone is becoming more feasible. PMID- 12541204 TI - Vertebroplasty in osteoporosis. AB - Percutaneous vertebroplasty (PV) is a safe and effective treatment for relieving pain in patients complaining of severe back pain induced by osteoporotic compression fractures. The success rate exceeds 90% and the complication rate is lower than 1%. Most of the complications are transient and should be avoided using good technique. The classic indication for PV is severe, persistent, and incapacitating focal back pain not responding to standard medical therapy and related to one or more collapsed vertebral bodies of 4 to 12 weeks duration. PV should be used earlier in patients at risk of immobilization complications and requiring narcotics. PV should be always considered as a good alternative treatment compared with medical therapy in patients with pain and osteoporotic compression fractures. PMID- 12541206 TI - Study of the motor corticospinal system in the developing rat fetus: comparison of Wistar and normal and hydrocephalic HTx rats. AB - The motor corticospinal system can be identified from day E14 in Wistar and HTx fetuses. There are no significant anatomical differences between the two species of rats. In addition, in day E17 Wistar and HTx fetuses cell counts in the cortical mantle (cortical plate, intermediate zone and germinal matrix) are similar. However, in day E20 fetuses there are significant differences in the number of cells in the cortical mantle of the hydrocephalic HTx fetuses compared to that in the Wistar and normal HTx fetuses, their total number of cells being reduced compared to that of the normal HTx and Wistars. Breakdown of the numbers of cells in the different layers shows that in the hydrocephalics there is a significant reduction in the number of cells in the germinal matrix and intermediate zone but, although the number of cells is also reduced in the cortical plate, the reduction is not significant. Measurements of the anterior/posterior width of the pyramid show that its growth is almost complete by day E17 and that on day E20 the measurements are similar in Wistar and normal and hydrocephalic HTx fetuses. These findings suggest that it is only cells generated after day E17 that are missing from the cortex of day E20 hydrocephalic rats. It is known that the motor corticospinal tract axons arise from pyramidal cells in layers 6, 5 and 4 of the cortical plate. These layers are generated earlier than layers 3 and 2 and are almost certainly in place by day E17 and account for why motor corticospinal tract function is spared in younger animals with established hydrocephalus. PMID- 12541207 TI - Ventricular shunting for hydrocephalus in children: patients, procedures, surgeons and institutions in English Canada, 1989-2001. AB - INTRODUCTION AND PURPOSE: Ventricular shunting remains the principle and most generally applicable method to treat hydrocephalus in children. This paper describes the demographics of this treatment in English Canada during the period of 1989 to March 2001. METHODS: Hospital discharge records were obtained for patients less than 18 years who had a shunt inserted or revised. A database was constructed relating patients and procedures to hospital discharges based on scrambled patient identifiers, year of birth, sex, postal code and diagnoses. OBSERVATIONS: 5,947 patients underwent ventricular shunting procedures for hydrocephalus in this period. 261 surgeons working in 73 institutions provided 12,106 interventions (Shunt insertions: ventriculoperitoneal--5009, ventriculoatrial--119, ventriculopleural--28. Revisions: 6,950). Infection was deemed to have occurred in 1,059 procedures. Over the study period, the median number of procedures performed per surgeon per year was 2, with 75 % of surgeons performing 5 or fewer procedures in children per year. Although many surgeons operated on children throughout the thirteen years of the study, many did not acquire substantive cumulative experience. Overall infection rate was 8.6 %. Surgeon infection rates were greater than or equal to 20 % during the first four years of practice and thereafter they fell to and remained in the 10 % range. The mean shunt survival at 12 months of individual surgeons varied between 50 - 60 %, regardless of the number of years of experience of the surgeon; however, performance variability as measured by the standard deviation of 12 month survival rates for all surgeons, adjusted for years of experience, ranged widely until the fifth year of practice. The average number of procedures per year for treating hospitals was 2 with 75 %, providing 12 or fewer services annually. Over the entire study, 50 % of institutions provided 10 or fewer procedures. The mean institutional infection rate was 11.4 % (SD 23, median--6.0). CONCLUSIONS: Quality monitoring of infection rate and duration of shunt function remains critical as many surgeons and hospitals provide care to children with hydrocephalus infrequently. Variability in infection rates and shunt survival at 12 months are a function of surgeon experience, measured by years in practice. Variability in outcome decreases with increasing surgeon experience. PMID- 12541208 TI - Chiari-II malformation: a biometric analysis. AB - INTRODUCTION: Chiari-II malformation is a complex set of anatomic abnormalities of the posterior fossa, brainstem, and cerebellum seen in myelomeningocele. Previous studies have tried to understand this condition and its clinical consequences by studying a few fixed anatomic landmarks as dependent variables and clinical symptoms as outcome variables. These studies have led to conflicting and limited results. As a first step towards a more comprehensive study of the structural-functional relationships in Chiari-II malformation, we undertook a biometric case-series analysis of a group of individuals with MMC, using both traditional methods to study size, and thin-plate spline methodology to study shape. MATERIALS AND METHODS: The traditional analysis was a retrospective, randomly obtained case-series of 25 individuals with myelomeningocele and Chiari II malformation. MRIs were digitized for biometric analysis of the component structures and compared to 25 controls. Landmarks on the digitized images were measured using Scion Image. Data were statistically analyzed using SPSS. The thin plate spline analysis used a subset of these individuals (n = 14). Landmarks on digitized MRIs were identified and marked with TPSDig software. Landmark data was then imported into Splus to generate a series of shape variables. Statistical analysis using Splus was then undertaken. RESULTS: Traditional analysis revealed relationships between vermian herniation and pontomedullary junction position (p < 0.05) as well as between the height (and overall size) of the posterior fossa and degree of vermian herniation (p < 0.05). The degree of brainstem herniation/elongation did not correlate with the degree of vermian herniation. The shape analysis revealed that there is as much shape variability within the group of Chiari-II patients as there is between this group and unaffected controls. Thin-plate spline analysis shows that the position of the caudal-most aspect of the vermis is the point of greatest brain rearrangement; likewise, the opisthion is the point of greatest bony rearrangement. Partial least-squares analysis shows that in Chiari-II patients, a strong correlation (r = 0.821) exists between the extent to which the brain is warped (specifically, in regard to vermian descent) and the extent to which the bone is warped (specifically, in regard to the displacement of the opisthion) (p < 0.05). CONCLUSION: Traditional biometric analysis of size confirms that the degree of vermian herniation and cervicomedullary junction herniation are independent variables in Chiari-II malformation. Posterior fossa size is an important factor in explaining the variability of vermian herniation. The relationship between IPN position and vermian herniation suggests the possibility of a common etiology. The phenomenon of cervicomedullary junction descent appears to be independent, suggesting a different etiology. Analysis of shape indicated that shape changes in bone and brain are related, and that the Chiari-II-associated abnormalities vary greatly in their degree of abnormality. The extreme morphological variability argues against the use of mean differences as a statistical technique in analyzing morphological abnormalities in the structures we investigated. PMID- 12541209 TI - Reduction of latex sensitisation in spina bifida patients by a primary prophylaxis programme (five years experience). AB - Spina bifida patients represent a group with the highest risk for latex sensitisation and allergy with life-threatening symptoms mostly during surgery. At the end of 1995 we initiated a primary latex prophylaxis around and during surgery and anaesthesia of all spina bifida patients. The aim of our study was to investigate the prevalence of latex sensitisation in the spina bifida patients born during the five years after establishing latex prophylaxis in the Cologne Children's Hospital in December 1995. We investigated 34 serum samples of 27 spina bifida patients (mean age 2.4 years) for specific IgE antibodies against latex allergens (CAP system) and compared these patients born after 1995 with 38 spina bifida patients up to 5 years of age (mean 3.1 years) born before. In the prophylaxis group two of 27 patients (7 %), one of them with two operations outside the Children's Hospital, had low specific IgE against latex ( 100 kU/l (mean 22.6 kU/l, min 0.4 kU/l). Sera of 22 patients remained negative for latex IgE (min. 1, max. 19, mean 4.3 operations). By primary latex prophylaxis during surgery, anaesthesia and in paediatric wards the prevalence of latex sensitisation can be significantly reduced even in the high risk group of spina bifida patients. Problems can arise by the need for surgery in hospitals not experienced in the treatment of spina bifida patients, where latex prophylaxis is neglected. PMID- 12541210 TI - Fetal surgery for spina bifida aperta: to be or not to be? AB - Fetal SBA (spina bifida aperta) has been operated upon in more than 213 cases in the USA in the last 8 years. Indications and results still remain controversial. Our group reproduced successfully Meuli's model in sheep and reviewed the literature on SBA and the Internet updated sites on the theme. Then we reviewed the follow-up of all cases operated upon postnatally in 20 years at a Neurosurgery Department in Rome. We published all the data regarding the three above-mentioned steps. This paper contains reflections on the three, and our opinion on the correct approach to prenatally diagnosed SBA, as far as fetal surgery is concerned. PMID- 12541211 TI - Ultrasound-guided needle aspiration of cranial epidural hematoma in a neonate: treating a rare complication of vacuum extraction. AB - Epidural hematoma is a rare form of neonatal birth injury accounting for 2% of newborn intracranial hemorrhage. We report the first case of ultrasound-guided needle aspiration of a cranial epidural hematoma in a neonate who also suffered subgaleal and intraparenchymal hemorrhage as a complication of vacuum extraction. PMID- 12541212 TI - Successful treatment of primary fetal hydrothorax with a double basket catheter. AB - Fetal pleural effusions can sometimes be detected before birth with ultrasonography. Intervention may be warranted when there is a condition that results in hydroplastic lung and/or fetal hydrops. A 22-week-old fetus with a severe pleural effusion and hydrops was successfully treated by long-term pleural drainage with a double basket catheter from 22 to 39 weeks of gestation. PMID- 12541213 TI - Gentamicin and tobramycin in neonates: comparison of a new extended dosing interval regimen with a traditional multiple daily dosing regimen. AB - Because of a lack of data supporting traditional dosing regimens for aminoglycosides, especially in extremely low-birth-weight infants, the authors developed revised dosing guidelines. The new guidelines increased doses to 5 mg/kg (over traditional doses of 2.5 mg/kg) and lengthened the dosing interval. When results of the two regimens were compared in 120 infants, 26.8% of infants in the traditional dosing group had subtherapeutic levels at <5 microg/mL, whereas only 1.3% of infants in the new practice dosing group were subtherapeutic. With the new dosing practice, serum levels in 1.3% of infants also exceeded the upper therapeutic range of 12 microg/mL. In conclusion, by increasing the dose of aminoglycosides and extending the dosing intervals, therapeutic levels-as defined by a C min <2 microg/mL and a C max of 5 to 12 microg/mL--were obtained significantly more often. In essence the regimen involves once daily dosing for infants <1200 g who are >30 days of age and for infants <1200 g who are >7 days of age. Serum concentrations still need to be monitored where clinically indicated. PMID- 12541214 TI - Doppler flow velocimetry assisted diagnosis of an intrauterine synechia during pregnancy. AB - The differential diagnosis of reflective intrauterine membranes during pregnancy includes amniotic bands of fetal origin or membranes of combined maternal-fetal origin. While the former have been associated with fetal transverse reduction defects or the lethal amniotic band syndrome, the latter are usually benign and consist of a preexisting uterine septation or synechia around which the fetal membranes fold or become enveloped. We present an unusual case in which a patient was noted at 19 weeks' gestation to have an intrauterine membrane of undetermined origin, appearing to contain a pulsating vessel. Color Doppler imaging and Doppler flow velocimetry clearly depicted a pulse synchronous with the maternal heart rate, defining the membrane as amniotic membranes surrounding a preexisting uterine synechia and not a true amniotic band. The pregnancy was otherwise uneventful and the patient delivered a healthy neonate at elective repeat cesarean at which time the presence of the uterine synechia was confirmed. PMID- 12541215 TI - Apoptosis of cord blood neutrophils and their response to colony-stimulating factors. AB - Neutrophil production and functions are immature in newborns. Although neutrophil kinetics during neonatal period have been widely studied, little is known about the effect of apoptosis on these defects. In this study, we examine the apoptosis of neonatal neutrophils and the effects of colony-stimulating factors (CSF) on this process. The study was performed using three different methodologies (morphological analysis, surface Fas expression, and mitochondrial 7A6 antigen expression) and the results were compared with adult controls. Neonatal neutrophils more rapidly underwent apoptosis in comparison to adult neutrophils. The above-mentioned three different methods gave similar results. Granulocyte-CSF (G-CSF) and granulocyte-macrophage CSF (GM-CSF) decreased the apoptosis of neutrophils in newborns and adults. This effect was significantly more pronounced in adults than newborns in morphological analysis. Increased apoptosis may contribute to qualitative and quantitative defects of neutrophils during neonatal period and may be an explanation for the proneness of newborn to develop neutropenia during systemic infections. PMID- 12541216 TI - Parotid swelling in a premature neonate. AB - Suppurative parotitis in neonates is very rare. We report a case of unilateral suppurative parotitis in a 29-week gestation age infant, who recovered completely following medical treatment. Diagnosis was by clinical examination and microbiology, and parotid involvement confirmed by computed tomographic (CT) scan. The causative organism was coagulase negative Staphylococcus epidermidis. PMID- 12541217 TI - Changes in skin temperature of hyperbilirubinemic newborns under phototherapy: conventional versus fiberoptic device. AB - The purpose of this study is to determine the changes in skin temperature of hyperbilirubinemic term newborns under conventional and fiberoptic phototherapy. The study included a group of 41 hyperbilirubinemic, but otherwise healthy, term infants, all of appropriate size for gestational age. The study was devised to include two separate groups: group 1 of 21 infants (51.2%) received conventional phototherapy, and group 2 of 20 infants (48.8%) received fiberoptic phototherapy. In both groups the surface temperature on the forehead, abdomen, left leg and back was calculated by infrared radiation thermometer (Cyclops Compac 3, Minolta, Land, England). A "Photo-Therapie 800 Heraeus" unit (Drager, Lubeck, Germany) was used for conventional phototherapy (CPT). A "Biliblanket PT system" (Ohmeda, Louisville, KY) was used for fiberoptic phototherapy (FO-PT). In our study we did not find significant differences of skin temperature in the four areas examined in the two groups prior to phototherapy. During conventional phototherapy, mean values of skin temperature were found to be significantly higher than those found before phototherapy. During fibreoptic phototherapy no statistically significant temperature differences were found on the forehead, abdomen, leg, and back before and during phototherapy. Furthermore, a statistically significant increase in skin temperature was found during phototherapy on the forehead, abdomen, leg, and back in patients of group 1 with respect to patients of group 2. We demonstrated that fibreoptic phototherapy, in contrast to conventional phototherapy, does not induce a significant increase in skin temperature. PMID- 12541218 TI - Maternal Arnold-Chiari type I malformation and syringomyelia: a labor management dilemma. AB - Arnold-Chiari type I malformations consists of elongation of the cerebellar tonsils with their displacement below the foramen magnum. Syringomyelia is an associated cyst that accumulates cerebrospinal fluid in the cord that can impinge on local nerve fibers. Pregnant women with either of these disorders are of special concern due to the potential risk of brain stem herniation and or spinal column compression from physiological changes that occur during labor. We present two cases. The first case is a patient with syringomyelia who was admitted in labor with worsening peripheral neurological symptoms. Epidural anesthesia was placed and she underwent an uncomplicated cesarean delivery with resolution of her symptoms postpartum. The second case is a patient with an Arnold-Chiari type I malformation and syringomyelia who presented in labor. The patient had an epidural placed and was allowed to progress to complete dilation and effacement at +2 station. She underwent a successful operative vaginal delivery without voluntary maternal expulsive efforts. Both patients had uncomplicated postpartum courses. Although these are rare disorders with significant potential morbidity, labor can be managed by either mode of delivery with careful patient selection. We caution that this review has insufficient numbers of patients to address the safety and efficacy of either delivery mode but rather focuses on alternatives for delivery. This report is the first to document a case of a patient with an Arnold-Chiari malformation and syringomyelia successfully managed in labor with a vaginal delivery. PMID- 12541219 TI - Reproductive risk factors, Doppler findings, and outcome of affected births in placental abruption: a population-based analysis. AB - Placental abruption complicates about 1% of all singleton pregnancies and the aim of this study is to assess the reproductive maternal risk factors associated with placental abruption, and the outcome of affected births. We analyze 170 women with singleton pregnancies complicated by placental abruption who gave birth at Kuopio University Hospital from March 1989 to December 1999. The general obstetric population ( n = 22,905) was selected as the reference group and logistic regression analysis was used to identify independent reproductive risk factors. Furthermore, Doppler ultrasonographic results and pregnancy outcome measures in the two groups were also recorded. The incidence of placental abruption was 0.57% in the referral area. Preeclampsia, grand multiparity, velamentous umbilical cord insertion, cigarette smoking, prior fetal demise, advanced maternal age (>35 years), and previous miscarriage were independent risk factors of placental abruption, with adjusted relative risks of 4.39, 3.60, 2.53, 2.46, 2.02, 1.62, and 1.55, respectively. Most cases of placental abruption occur before the onset of labor in low-risk pregnancies and are not predictable with regard to maternal reproductive risk factors. Current antepartum methods of detecting uteroplacental problems, including Doppler ultrasonography, are not effective in prenatal prediction of placental abruption. The outcome of affected births is still poor. PMID- 12541221 TI - MRI measurement of time-resolved wall shear stress vectors in a carotid bifurcation model, and comparison with CFD predictions. AB - PURPOSE: To study pulsatile fluid flow in a physiologically realistic model of the human carotid bifurcation, and to derive wall shear stress (WSS) vectors. MATERIALS AND METHODS: WSS vectors were calculated from time-resolved 3D phase contrast (PC) MRI measurements of the velocity field. The technique was first validated with sinusoidal flow in a straight tube, and then used in a model of a healthy human carotid bifurcation. Velocity measurements in the inflow and outflow regions were also used as boundary conditions for computational fluid dynamics (CFD) calculations of WSS, which were compared with those derived from MRI alone. RESULTS: The straight tube measurements gave WSS results that were within 15% of the theoretical value. WSS results for the phantom showed the main features expected from fluid dynamics, notably the low values in the bulb region of the internal carotid artery, with a return to ordered flow further downstream. MRI was not able to detect the high WSS values along the divider wall that were predicted by the CFD model. Otherwise, there was good general agreement between MRI and CFD. CONCLUSION: This is the first report of time-resolved WSS vectors estimated from 3D-MRI data. The technique worked well except in regions of disturbed flow, where the combination with CFD modeling is clearly advantageous. PMID- 12541220 TI - Sensitive and efficient detection of RB1 gene mutations enhances care for families with retinoblastoma. AB - Timely molecular diagnosis of RB1 mutations enables earlier treatment, lower risk, and better health outcomes for patients with retinoblastoma; empowers families to make informed family-planning decisions; and costs less than conventional surveillance. However, complexity has hindered clinical implementation of molecular diagnosis. The majority of RB1 mutations are unique and distributed throughout the RB1 gene, with no real hot spots. We devised a sensitive and efficient strategy to identify RB1 mutations that combines quantitative multiplex polymerase chain reaction (QM-PCR), double-exon sequencing, and promoter-targeted methylation-sensitive PCR. Optimization of test order by stochastic dynamic programming and the development of allele-specific PCR for four recurrent point mutations decreased the estimated turnaround time to <3 wk and decreased direct costs by one-third. The multistep method reported here detected 89% (199/224) of mutations in bilaterally affected probands and both mutant alleles in 84% (112/134) of tumors from unilaterally affected probands. For 23 of 27 exons and the promoter region, QM-PCR was a highly accurate measure of deletions and insertions (accuracy 95%). By revealing those family members who did not carry the mutation found in the related proband, molecular analysis enabled 97 at-risk children from 20 representative families to avoid 313 surveillance examinations under anesthetic and 852 clinic visits. The average savings in direct costs from clinical examinations avoided by children in these families substantially exceeded the cost of molecular testing. Moreover, health care savings continue to accrue, as children in succeeding generations avoid unnecessary repeated anaesthetics and examinations. PMID- 12541222 TI - MR imaging of the thyroid: correlation between apparent diffusion coefficient and thyroid gland scintigraphy. AB - PURPOSE: To evaluate the usefulness of echo-planar MR imaging for assessing the thyroid function and confirm the clinical use of MR imaging for thyroid diseases. MATERIALS AND METHODS: Thirty-four patients with a variety of thyroid disorders (24 Graves disease; five subacute thyroiditis; five Hashimoto thyroiditis) were examined using T1-, T2-, and diffusion-weighted magnetic resonance (MR) imaging and thyroid scintigraphy with Tc-99m pertechnetate. RESULTS: The ADC values obtained from the diffusion-weighted images of the patients with Graves disease were significantly higher than those of patients with subacute hyroiditis and Hashimoto thyroiditis, though no difference among those disorders was observed on T1- and T2-weighted images. Based on the ADC value, anisotropy was not observed in the thyroid gland. An ADC value of 1.82 x 10(-3) mm(2)/second or higher indicated the presence of Graves disease (sensitivity 75%, specificity 80%). CONCLUSION: Diffusion-weighted MR images may be of value for the diagnosis of thyroid diseases and could be clinically important in the evaluation of thyroid function. PMID- 12541223 TI - Coronary MR angiography: respiratory motion correction with BACSPIN. AB - PURPOSE: To improve the signal-to-noise ratio (SNR) of breath-held coronary magnetic resonance angiography (CMRA) without increasing the number or duration of breath holds. MATERIALS AND METHODS: In this BACSPIN (Breathing AutoCorrection with SPiral INterleaves) technique, a single breath-held electrocardiogram (ECG) gated multi-slice interleaved-spiral data set is acquired, followed by repeated imaging of the same slices during free breathing. Each spiral interleaf from the breath-held data set is used as a standard for comparison with corresponding acquisitions at the same interleaf angle during free breathing. The most closely matched acquisitions are incorporated into a multi-slice, multi-average data set with increasing SNR over time. In-plane translations of the coronary artery can be measured and compensated for each accepted acquisition before combination with the other acquisitions. RESULTS: CMRA was performed on six volunteers, with improved SNR and minimal motional blurring. In some cases, breath holding could be dispensed with completely and the average respiratory position used as a reference. CONCLUSION: BACSPIN provides a promising method for CMRA with improved SNR and limited breath-holding requirements. PMID- 12541224 TI - MR findings of collateral circulation are more accurate measures of hemodynamic significance than arm-leg blood pressure gradient after repair of coarctation of the aorta. AB - PURPOSE: To determine the relationship between percent stenosis and three indicators of hemodynamic significance-arm-leg blood pressure gradient, direct visualization of collaterals, and percent increase flow from proximal to distal descending thoracic aorta-in patients with prior repair of coarctation of the aorta (CoA). MATERIALS AND METHODS: Magnetic resonance imaging (MRI) examinations of 19 patients with prior repair of CoA were retrospectively reviewed. Percent stenosis was compared to the arm-leg blood pressure gradient as obtained from chart review, the depiction of collaterals by gadolinium-enhanced magnetic resonance angiography (MRA), and the percent increased flow in the distal thoracic aorta as measured by velocity-encoded cine MRI. Some imaging series or blood-pressure values were not available in some patients. All of the data were available for 15 of the 19 patients. RESULTS: The arm-leg blood pressure gradient showed no statistical association with percent stenosis (R(2) = 0.10, P = 0.22), direct visualization of collaterals (P = 0.80), or percent increase in flow (R(2) < 0.01, P = 0.85). Percent stenosis did show association with visualization of collaterals (P = 0.01) and increase flow (R(2) = 0.62, P < 0.01). CONCLUSION: The arm-leg blood pressure gradient is not a reliable indicator of hemodynamic significance of restenosis in patients with prior repair of CoA. Direct visualization of collateral vessels by MRA and percent increase in flow from proximal to distal descending thoracic aorta are reliable indicators of hemodynamic significance. PMID- 12541225 TI - Serial, noninvasive, in vivo magnetic resonance microscopy detects the development of atherosclerosis in apolipoprotein E-deficient mice and its progression by arterial wall remodeling. AB - PURPOSE: To test the ability of serial, in vivo magnetic resonance microscopy (MRM) to detect the development of atherosclerosis and quantify its progression in apolipoprotein E-deficient mice. MATERIALS AND METHODS: The abdominal aortae of six ApoE(-/-) and three wild-type (WT) control mice were imaged by MRM at 9.4T. Proton density weighted images were obtained (TR = 2000, TE = 9 msec) using four signal averages. The image resolution was 109 x 109 x 500 microm(3). The six ApoE(-/-) mice underwent serial MRM three to five times over a period < or = 44 weeks. Multiple, anatomically aligned MRM slices (N = 6-11 per time point, total 202) were compared serially in each animal. RESULTS: The abdominal aorta remained free of atherosclerosis until 20 weeks of age but thereafter, atherosclerosis was identified in all ApoE(-/-) mice (P < 0.05 to P < 0.001), but no WT controls. Lesion progression was accompanied by positive remodeling in which atherosclerosis within the aortic wall was accommodated by an increase in total cross sectional area (P < 0.01), while lumen area was unchanged. CONCLUSION: Serial MRM demonstrated the development and progression of atherosclerosis in mouse aorta. Importantly, progression of atherosclerosis could be identified within individual animals. By following the same aortic lesions over time, MRM demonstrated that progression of atherosclerosis in mice is associated with positive arterial remodeling. PMID- 12541226 TI - MRI of the liver: can True FISP replace HASTE? AB - PURPOSE: To determine the diagnostic accuracy of two fast breath-hold magnetic resonance (MR) imaging sequences, half-Fourier acquired single turbo spin-echo (HASTE) and true fast imaging with steady state precession (TrueFISP), for the detection and characterization of focal liver lesions MATERIALS AND METHODS: A total of 186 patients with suspected focal liver lesions were enrolled in this study. All patients underwent the same standardized study protocol including HASTE and TrueFISP. A consensus reading based on all available image data served as a standard of reference for classifying lesions into cysts, hemangiomas, focal nodular hyperplasia, or malignant/other lesions. All malignant lesions, as well as hepatic adenomas and abscesses, were histologically verified. Each separated by an eight-week interval, HASTE and TrueFISP images were retrospectively reviewed in random order for the detection and characterization of focal hepatic lesions. Finally, a receiver operating characteristic (ROC) analysis was calculated. RESULTS: HASTE images had an overall sensitivity of 0.86 and a specificity of 0.91, whereas TrueFISP showed an overall sensitivity and specificity of 0.79 and 0.83, respectively (p>0.1). CONCLUSION: Neither HASTE nor TrueFISP alone are sufficient for the detection and characterization of hepatic lesions. PMID- 12541227 TI - Feasibility of abdomino-pelvic T1-weighted real-time thermal mapping of laser ablation. AB - PURPOSE: To prove the hypothesis that T1-weighted (T1W) thermal mapping is reliable and achievable in magnetic resonance (MR)-guided laser tumor ablation. MATERIALS AND METHODS: Near real-time gray and color-scale T1W thermal maps in 111 MR-guided laser thermal ablations (LTA) of liver, kidney, and uterine tumors were studied. After laser fiber placement, near real-time gray and color-scale thermal maps were produced. Previous work showed T1 signal is inversely proportional to temperature below 55 degrees C (the point of irreversible necrosis). RESULTS: Thermal mapping was successful in 84% of uterine, 74% of hepatic, and 20% of renal ablations. For hepatic and uterine tumors, size and conspicuity of thermal lesions were significantly greater on subtracted colorized images rather than gray-scale raw image mapping (P = 0.001, paired Student's t test). Patient movement (N = 24), fiber charring N = 2), magnetic field distortion, and reconstruction errors (N = 2) caused mapping failure. For both renal and hepatic tumors, comparison of near real-time T1W colorized thermal map and follow up gadolinium-enhanced MR imaging revealed moderate correlation (Pearson correlation coefficient of 0.55 and 0.5, respectively). CONCLUSION: In hepatic, renal, and uterine thermal maps, the color scale produced significantly greater sized lesions with significantly greater conspicuity than the gray scale. T1W thermal mapping was reliable and successfully achieved in 73.7% of procedures, and predicted the ablated area of the tumor moderately well. PMID- 12541228 TI - Precise 3D skeletal kinematics using fast phase contrast magnetic resonance imaging. AB - PURPOSE: To examine the precision of cine-phase contrast (PC) magnetic resonance imaging (MRI) techniques as applied to the quantification of three-dimensional knee joint kinematics. MATERIALS AND METHODS: The knee joints of eight healthy volunteers were studied using three different dynamic, PC MRI protocols: cine-PC (one average), cine-PC (two averages), and cine-PC with segmented phase encoding (fast-PC). RESULTS: Fast-PC has comparable precision, shorter scan times, and improved subject interexam variability (SIEV) compared to cine-PC (two averages). Further, cine-PC (one average) has low precision and high SIEV, making fast-PC the preferred method of data acquisition. Specifically, the precision of fast-PC MRI in measuring knee joint kinematics ranged from 0.22 degrees -1.16 degrees. CONCLUSION: A cine-PC MRI technique utilizing segmented phase encoding (fast-PC MRI) acquires dynamic data at a faster rate than other PC imaging protocols, without compromising data precision. Being able to acquire precise 3D kinematics with shorter imaging times is critical if we are to use this technique to advance ongoing research in musculoskeletal kinematics. PMID- 12541229 TI - Percutaneous MR-guided discography in a low-field system using optical instrument tracking: a feasibility study. AB - PURPOSE: To evaluate the feasibility of MRI-guided discography with optical tracking. MATERIAL AND METHODS: 12 consecutive patients who had a clinical suspicion of lumbar discogenic pain and/or suggestive finding of disc degeneration in imaging studies (MRI, CT, plain radiography) underwent MRI-guided discography in order to determine possible pain provocation during puncture and contrast injection. An 0.23 T open configuration MRI device with interventional tools (Outlook Proview, Philips Medical Systems, MR Technologies, Finland) was used in procedural imaging and instrument guidance. An optical guidance tool was attached to the MRI compatible needle (Chiba-type MReye, Cook, Bloomington, IN). After initial disc puncture, 1-2 mL of gadolinium contrast (Magnevist, 469 mg/mL, Schering AG, Germany) saline mixture (1:8) was injected into the disc. Immediately after injection, sagittal FE T1 weighted images were obtained to verify the final position of the needle and formation of the MRI discogram. On nine patients, additional noncontrast sagittal fast spin echo (FSE) T1, FSE T2, and axial 3D T1 gradient echo imaging was performed before and after contrast media injection to obtain MRI discograms. RESULTS: Overall, 35 disc punctures were initialized and 34 MRI discograms were obtained. In all punctures, a positive or negative pain response was obtained. The average time for performing a procedure for three discs was 1 hour 25 minutes (minimum 45 minutes, maximum 2 hours, 15 minutes), and the average number of imaging sequences used for a puncturing one disc was 12. On one disc, the puncture failed and a discogram was not acquired. There was one complication (disc collapse) reported during follow up. CONCLUSION: Our results show that MRI guidance in performing discography is accurate and relatively safe. It is a technically comparable method to CT guidance or fluoroscopy. PMID- 12541230 TI - 8.0-Tesla human MR system: temperature changes associated with radiofrequency induced heating of a head phantom. AB - PURPOSE: To investigate if the heat induced in biological tissues by typical radio frequency (RF) energy associated with an 8.0-Tesla magnetic resonance (MR) system causes excessive temperature changes. MATERIALS AND METHODS: Fluoroptic thermometry was used to measure temperatures in multiple positions in a head phantom made of ground turkey breast. A series of experiments were conducted with measurements obtained at RF power levels ranging from a specific absorption rate (SAR) of up to 4.0 W/kg for 10 minutes. RESULTS: The highest temperature increases were up to 0.7 degrees C. An inhomogeneous heating pattern was observed. In general, the deep regions within the phantom registered higher temperature increases compared to the peripheral sites. CONCLUSION: The expectation of an inhomogeneous RF distribution in ultra high field systems (> 4 T) was confirmed. At a frequency of 340 MHz and in-tissue RF wave length of about 10 cm, the RF inhomogeneity was measured to create higher temperatures in deeper regions of a human head phantom compared to peripheral tissues. Our results agree with the computational electromagnetic calculations for such frequencies. Importantly, these experiments indicated that there were no regions of heating that exceeded the current FDA guidelines. PMID- 12541231 TI - Variation correction algorithm: analysis of phase suppression and thermal profile fidelity for proton resonance frequency magnetic resonance thermometry at 0.2 T. AB - PURPOSE: To develop and analyze the performance of the variation correction algorithm (VCA), a phase correction technique that mitigates the contribution of background phase variations by combining accurate alignment of echoes, K-space based phase correction (as opposed to spatial polynomials), and extraction of alias-free phase difference images. MATERIALS AND METHODS: A series of echo shifted gradient-recalled echo (GRE) images was processed with K-space alignment and phase corrected with increasing sizes of M x M masks of central K-space coefficients. The extent of background phase variation suppression due to magnet field drift was assessed. Further, a simulated thermal profile was superimposed on the same data in a related experiment. Residual errors in reconstructed simulated thermal profiles were quantitatively characterized to estimate algorithm performance. RESULTS: Using a 3 x 3 K-space mask, the VCA was able to 1) maintain the typical mean background error in a 35 x 35 pixel region of interest (ROI) at -0.1 degrees C; and 2) reconstruct, relative to the applied thermal profile, a phase-corrected profile that typically contains a 1.7 degrees C underestimation of peak temperature difference and a mean error along the 60 degrees C line of -0.8 degrees C. CONCLUSION: The results suggest that thermal profiles can be accurately reconstructed at 0.2 T using the VCA, even in the presence of over 1 ppm spatially and temporally dependent field drift over a 1 hour time frame. PMID- 12541232 TI - Can dynamic susceptibility contrast magnetic resonance imaging perfusion data be analyzed using a model based on directional flow? AB - PURPOSE: To examine the implications of a physiological model of cerebral blood that uses the contradictory assumption that blood flow in all voxels of DSCE-MRI data sets is directional in nature. Analysis of dynamic susceptibility contrast enhanced magnetic resonance imaging (DSCE-MRI) uses techniques based on indicator dilution theory. Underlying this approach is an assumption that blood flow through pixels of gray and white matter is entirely random in direction. MATERIALS AND METHODS: We have used a directional flow model to estimate theoretical blood flow velocities that would be observed through normal cerebral tissues. Estimates of flow velocities from individual pixels were made by measuring the mean transit time for net flow (nMTT). Measurements of nMTT were made for each voxel by estimating the mean difference in contrast arrival time between each of the adjacent six voxels. RESULTS: Examination of the spatial distribution of contrast arrival time from DSCE-MRI data sets in normal volunteers demonstrated clear evidence of directional flow both in large vessels and in gray and white matter. The mean velocities of blood flow in gray and white matter in 12 normal volunteers were 0.25 +/- 0.013 and 0.21 +/- 0.014 cm/second, respectively, compared to predicted values of 0.25 and 0.18 cm/second. These values give measured nMTT for a 1-mm isotropic voxel of gray and white matter of 0.45 +/- 0.12 and 0.52 +/- 0.11 seconds, respectively, compared to predicted values of 0.47 and 0.55 seconds. CONCLUSION: A directional model of blood flow provides an alternative approach to the calculation of cerebral blood flow from (CBF) DSCE-MRI data. PMID- 12541233 TI - In vivo and in vitro MR spectroscopic profile of central neurocytomas. AB - This study reports in vivo and in vitro magnetic resonance spectroscopic findings in two cases of central neurocytomas (CNC) confirmed by immunohistochemistry. Volume localized in vivo proton magnetic resonance spectroscopy (MRS) was carried out before surgery using a point resolved spectroscopy (PRESS) sequence with a repetition time of six seconds and an echo time of 135 msec. Normal spectrum was obtained from gray matter from a volunteer for comparison. (1)H and (31)P in vitro MRS studies were carried out at 9.4 T on the extracts prepared from the surgically excised tumors. The in vivo spectra showed prominent glycine (Gly) and choline (Cho) and low N-acetyl aspartate compared to the normal. The Gly peak was assigned using the in vitro studies. These studies showed that the major contribution to the Cho peak observed in vivo is from phosphocholine. A combination of the presence of NAA and an increased Gly in the proton MR spectrum could be a characteristic feature of CNCs, which are rare intraventricular tumors of neuronal origin. PMID- 12541234 TI - Multispectral magnetic resonance image analysis using principal component and linear discriminant analysis. AB - PURPOSE: To explore the possibilities of combining multispectral magnetic resonance (MR) images of different patients within one data matrix. MATERIALS AND METHODS: Principal component and linear discriminant analysis was applied to multispectral MR images of 12 patients with different brain tumors. Each multispectral image consisted of T1-weighted, T2-weighted, proton-density weighted, and gadolinium-enhanced T1-weighted MR images, and a calculated relative regional cerebral blood volume map. RESULTS: Similar multispectral image regions were clustered, while dissimilar multispectral image regions were scattered in a single plot. Both principal component and linear discriminant analysis allowed discrimination between healthy and tumor regions on the image. In addition, linear discriminant analysis allowed discrimination between oligodendrogliomas and astrocytomas. However, the discriminant analysis method was partially capable of recognizing the tumor identity in unknown multispectral images. CONCLUSION: The proposed method may help the radiologist in comparing multispectral MR images of different patients in a more easy and objective way. PMID- 12541235 TI - Efficacy of passive acoustic screening: implications for the design of imager and MR-suite. AB - PURPOSE: To investigate the efficacy of passive acoustic screening in the magnetic resonance (MR) environment by reducing direct and indirect MR-related acoustic noise, both from the patient's and health worker's perspective. MATERIALS AND METHODS: Direct acoustic noise refers to sound originating from the inner and outer shrouds of the MR imager, and indirect noise to acoustic reflections from the walls of the MR suite. Sound measurements were obtained inside the magnet bore (patient position) and at the entrance of the MR imager (health worker position). Inner and outer shrouds and walls were lined with thick layers of sound insulation to eliminate the direct and indirect acoustic pathways. Sound pressure levels (SPLs) and octave band frequencies were acquired during various MR imaging sequences at 1.5 T. RESULTS: Inside the magnet bore, direct acoustic noise radiating from the inner shroud was most relevant, with substantial reductions of up to 18.8 dB when using passive screening of the magnetic bore. At the magnet bore entrance, blocking acoustic noise from the outer shroud and reflections showed significant reductions of 4.5 and 2.8 dB, respectively, and 9.4 dB when simultaneously applied. Inner shroud coverage contributed minimally to the overall SPL reduction. CONCLUSION: Maximum noise reduction by passive acoustic screening can be achieved by reducing direct sound conduction through the inner and outer shrouds. Additional measures to optimize the acoustic properties of the MR suite have only little effect. PMID- 12541236 TI - Are time-intensity curves helpful in differentiating between benign vertebral compression fracture and vertebral metastasis? PMID- 12541238 TI - In vivo 31P magnetic resonance spectroscopy of human brain at 7 T: an initial experience. AB - In vivo (31)P spectra were acquired from the human primary visual cortex at 7 T. The relaxation times of the cerebral metabolites, intracellular pH, rate constant (k(f)) of the creatine kinase (CK) reaction, and nuclear Overhauser enhancement (NOE) on the detected phosphorus moieties from irradiation of the water spins were measured from normal subjects. With a 5-cm-diameter surface coil, 3D (31)P chemical shift imaging was performed with a spatial resolution of 7.5 ml and an acquisition resolution of 8 min, resulting in a signal-to-noise ratio (SNR) for phosphocreatine (PCr) resonance of 32. The apparent T(1) and T(2) of PCr measured at 7 T were 3.37 +/- 0.29 s and 132.0 +/- 12.8 ms, respectively, which were considerably longer than those of adenosine triphosphate (ATP) (T(1): 1.02-1.27 s; T(2): 25-26 ms). The NOE measured in this study was 24.3% +/- 1.6% for PCr, and 10% for ATP. The k(f) measured in the human primary visual cortex was 0.24 +/ 0.03 s(-1). The results from this study suggest that ultra-high-field strength is advantageous for performing in vivo (31)P magnetic resonance spectroscopy (MRS) in the human brain. PMID- 12541239 TI - Oscillating gradient measurements of water diffusion in normal and globally ischemic rat brain. AB - Oscillating gradients were used to probe the diffusion-time/frequency dependence of water diffusion in the gray matter of normal and globally ischemic rat brain. In terms of a conventional definition of diffusion time, the oscillating gradient measurements provided the apparent diffusion coefficient (ADC) of water with diffusion times between 9.75 ms and 375 micros, an order of magnitude shorter than previously studied in vivo. Over this range, ADCs increased as much as 24% in vivo and 50% postmortem, depending on the nature of the oscillating gradient waveform used. Novel waveforms were employed to sample narrow frequency bands of the so-called diffusion spectrum. This spectral description of ADC includes the effects of restriction and/or flow, and is independent of experimental parameters, such as diffusion time. The results in rat brain were found to be consistent with restricted diffusion and the known micro-anatomy of gray matter. Differences between normal and postmortem data were consistent with an increase in water restriction and/or a decrease in flow, and tentatively suggest that physical changes following the onset of ischemia occur on a scale of about 2 microm, similar to a typical cellular dimension in gray matter. PMID- 12541240 TI - Undersampled projection reconstruction for active catheter imaging with adaptable temporal resolution and catheter-only views. AB - In this study undersampled projection reconstruction (PR) was used for rapid catheter imaging in the heart, employing steady-state free precession (SSFP) contrast. Active catheters and phased-array coils were used for combined imaging of anatomy and catheter position in swine. Real-time imaging of catheter position was performed with relatively high spatial and temporal resolution, providing 2 x 2 x 8 mm spatial resolution and four to eight frames per second. Two interactive features were introduced. The number of projections (Np) was adjusted interactively to trade off imaging speed and artifact reduction, allowing acquisition of high-quality or high-frame-rate images. Thin-slice imaging was performed, with interactive requests for thick-slab projection images of the signal received solely from the active catheter. Briefly toggling on catheter only projection images was valuable for verifying that the catheter tip was contained within the selected slice, or for locating the catheter when part of it was outside the selected slice. PMID- 12541241 TI - Metabolic profiles of human brain tumors using quantitative in vivo 1H magnetic resonance spectroscopy. AB - Proton spectroscopy can noninvasively provide useful information on brain tumor type and grade. Short- (30 ms) and long- (136 ms) echo time (TE) (1)H spectra were acquired from normal white matter (NWM), meningiomas, grade II astrocytomas, anaplastic astrocytomas, glioblastomas, and metastases. Very low myo-Inositol ([mI]) and creatine ([Cr]) were characteristic of meningiomas, and high [mI] characteristic of grade II astrocytomas. Tumor choline ([Cho]) was greater than NWM and increased with grade for grade II and anaplastic astrocytomas, but was highly variable for glioblastomas. Higher [Cho] and [Cr] correlated with low lipid and lactate (P < 0.05), indicating a dilution of metabolite concentrations due to necrosis in high-grade tumors. Metabolite peak area ratios showed no correlation with lipids and mI/Cho (at TE = 30 ms), and Cr/Cho (at TE = 136 ms) best correlated with tumor grade. The quantified lipid, macromolecule, and lactate levels increased with grade of tumor, consistent with progression from hypoxia to necrosis. Quantification of lipids and macromolecules at short TE provided a good marker for tumor grade, and a scatter plot of the sum of alanine, lactate, and delta 1.3 lipid signals vs. mI/Cho provided a simple way to separate most tumors by type and grade. PMID- 12541242 TI - The carnosine C-2 proton's chemical shift reports intracellular pH in oxidative and glycolytic muscle fibers. AB - The appearance of new peaks in the 7.7-8.6 and 6.8-7.4 ppm regions of the postexercise (1)H spectrum of frog muscle is reported. These new peaks result from the splitting of single pre-exercise carnosine C-2 and C-4 peaks into two peaks, representing the intracellular pH (pH(I)) of oxidative and glycolytic fibers. The following data support this conclusion: 1) comparison of means and regression analysis indicates equivalence of the pH(I) measurements by (1)H and (31)P NMR; 2) the pre- and poststimulation concentrations of carnosine are equal; 3) in ischemic rat hindlimb muscles, the presence of a single, more acidic peak in the plantaris; a single, less acidic peak in the soleus; and two peaks (more and less acidic) in the gastrocnemius correspond to published values for the fiber-type composition of these muscles; and 4) in muscles treated with iodoacetate prior to and during stimulation, a second peak never appears. These data indicate that it is feasible to measure separately the pH(I) of oxidative and glycolytic fibers using (1)H NMR spectroscopy. PMID- 12541243 TI - Quantitative HRMAS proton total correlation spectroscopy applied to cultured melanoma cells treated by chloroethyl nitrosourea: demonstration of phospholipid metabolism alterations. AB - Recent NMR spectroscopy developments, such as high-resolution magic angle spinning (HRMAS) probes and correlation-enhanced 2D sequences, now allow improved investigations of phospholipid (Plp) metabolism. Using these modalities we previously demonstrated that a mouse-bearing melanoma tumor responded to chloroethyl nitrosourea (CENU) treatment in vivo by altering its Plp metabolism. The aims of the present study were to investigate whether HRMAS proton total correlation spectroscopy (TOCSY) could be used as a quantitative technique to probe Plp metabolism, and to determine the Plp metabolism response of cultured B16 melanoma cells to CENU treatment in vitro. The exploited TOCSY signals of Plp derivatives arose from scalar coupling among the protons of neighbor methylene groups within base headgroups (choline and ethanolamine). For strongly expressed Plp derivatives, TOCSY signals were compared to saturation recovery signals and demonstrated a linear relationship. HRMAS proton TOCSY was thus used to provide concentrations of Plp derivatives during long-term follow-up of CENU-treated cell cultures. Strong Plp metabolism alteration was observed in treated cultured cells in vitro involving a down-regulation of phosphocholine, and a dramatic and irreversible increase of phosphoethanolamine. These findings are discussed in relation to previous in vivo data, and to Plp metabolism enzymatic involvement. PMID- 12541244 TI - Renal and systemic pH imaging by contrast-enhanced MRI. AB - Perturbations of renal and systemic pH accompany diseases of the kidney, such as renal tubular acidosis, and the ability to image tissue pH would be helpful to assess the extent and severity of such conditions. A dual-contrast-agent strategy using two gadolinium agents, the pH-insensitive GdDOTP(5-) and the pH-sensitive GdDOTA-4AmP(5-), has been developed to generate pH maps by MRI. The renal pharmacokinetics of the structurally dissimilar pH-insensitive contrast agents GdDTPA(2-) and GdDOTP(5-) were found to be similar. On that basis, and on the basis of similarity of structure and charge, the renal pharmacokinetics of GdDOTP(5-) and GdDOTA-4AmP(5-) were assumed to be identical. Dynamic T(1) weighted images of mice were acquired for 1 hr each following boluses of GdDOTP(5 ) and GdDOTA-4AmP(5-). The time-varying apparent concentration of GdDOTP(5-) and the time-varying enhancement in longitudinal relaxation rate following GdDOTA 4AmP(5-) were calculated for each pixel and used to compute pH images of the kidneys and surrounding tissues. MRI pH maps of control mice show acidic regions corresponding to the renal papilla, calyx, and ureter. Pretreatment of mice with the carbonic anhydrase inhibitor acetazolamide resulted in systemic metabolic acidosis and accompanying urine alkalinization that was readily detected by this dual-contrast-agent approach. PMID- 12541245 TI - MRI-guided coronary catheterization and PTCA: A feasibility study on a dog model. AB - The aim of this work was to demonstrate the feasibility of MRI-guided coronary artery catheterization and intervention in a dog model. Experiments were performed on 10 healthy dogs. A 9F introducer sheath was placed through a right carotid artery cutdown. A prototype 0.014-inch coronary MRI guidewire, a prototype 7 French MRI-guiding catheter, and two flexible surface coils were connected to a GE 1.5 T CV/i scanner for simultaneous visualization of the guidewire, guiding catheter, and chest anatomy. Images were displayed in real time on an in-room monitor. A nongated, single-slice fast gradient-echo sequence was used to obtain real-time images of the catheters and background anatomy during the intervention. Fifteen selective catheterizations were attempted in the coronary arteries, and all were successful. Selective injection of diluted gadolinium into the MRI-guiding catheter provided dynamic 2D projection coronary angiography in all cases, confirming successful catheterization. Percutaneous transluminal coronary angioplasty (PTCA) was attempted after two catheterizations, and all attempts were successful. Inflation of the balloon angioplasty catheter was performed successfully in the left anterior and circumflex arteries. Our results indicate that coronary artery catheterization and intracoronary balloon angioplasty are feasible with MRI guidance only. MRI guidance may be used as an alternative to X-ray guidance in coronary artery interventions in the future. PMID- 12541246 TI - Characterization of effects of mean arterial blood pressure induced by cocaine and cocaine methiodide on BOLD signals in rat brain. AB - A total of 45 male Sprague-Dawley rats were employed to determine whether cocaine or cocaine methiodide (CM) administration can induce a significant increase in mean arterial blood pressure (MABP) in rats, and whether such an increase in MABP can produce a global increase in blood oxygenation level-dependent (BOLD) contrast in the rat brain detectable by functional magnetic resonance imaging (fMRI). Cocaine methiodide is a quaternary derivative of cocaine that shares the same cardiovascular effects of cocaine, but does not penetrate the blood-brain barrier (BBB). Experimental results demonstrated that both CM (with doses of 2.5 and 7.5 mg/kg) and cocaine (with doses of 1.25 and 5.0 mg/kg) can induce a significant MABP change (30-80%). It was found that CM can only produce scattered, weak, and transient BOLD signals in a few voxels of the rat brain, and that these MABP-induced BOLD signals are not dose-dependent. In contrast, the administration of cocaine induced dose-dependent biphasic BOLD signals that were consistent with pharmacologically-induced cerebral vascular constriction and neuronal activity in the mesolimbic systems of the rat brain. The potential confounding factor of the MABP changes had little effect on the interpretation of drug-induced BOLD signal changes. These results confirm that the BOLD-weighted fMRI method can be extended to map drug-induced neuronal activity. PMID- 12541247 TI - Directional correlation characterization and classification of white matter tracts. AB - To study the architectural characteristics of white matter (WM) tracts, the directional correlation (DC), defined as the inner product of the major eigenvector of adjacent pixels, was used as a quantitative index to investigate directional similarity in WM tracts. A region-growing algorithm was employed to propagate an area from a seed point as a function of the DC threshold (DCt) to critically evaluate the directional properties of WM tracts. As the DCt was increased, more pixels were excluded from the propagated region as their DC fell below the DCt, and neighboring WM tracts could be distinguished as the area decreased. Taking the DC into account, a systematic classification routine for WM tracts was devised and tested on a mouse brain in vivo. The results show that individual WM tracts possess a high degree of directional similarity, and, by careful choice of the DCt value, the proposed classification algorithm can recognize all possible WM tracts in a given data set. PMID- 12541248 TI - Quantification of renal perfusion using an intravascular contrast agent (part 1): results in a canine model. AB - In this work absolute values of regional renal blood volume (rRBV) and flow (rRBF) are assessed by means of contrast-enhanced (CE) MRI using an intravascular superparamagnetic contrast agent. In an animal study, eight foxhounds underwent dynamic susceptibility-weighted MRI upon injection of contrast agent. Using principles of indicator dilution theory and deconvolution analysis, parametric images of rRBV, rRBF, and mean transit time (MTT) were computed. For comparison, whole-organ blood flow was determined invasively by means of an implanted flow probe, and the weight of the kidneys was evaluated postmortem. A mean rBV value of 28 ml/100 g was found in the renal cortex, with a corresponding mean rBF value of 524 ml/100 g/min and an average MTT of about 3.4 s. Although there was a systematic difference between the absolute blood flow values determined by MRI and the ultrasonic probe, a significant correlation (r(s) = 0.72, P < 0.05) was established. The influence of the arterial input function (AIF), T(1) relaxation effects, and repeated measurements on the precision of the perfusion quantitation is discussed. PMID- 12541249 TI - Quantification of renal perfusion abnormalities using an intravascular contrast agent (part 2): results in animals and humans with renal artery stenosis. AB - The interrelation between the morphologic degree of renal artery stenosis and changes in parenchymal perfusion is assessed using an intravascular contrast agent. In seven adult foxhounds, different degrees of renal artery stenosis were created with an inflatable clamp implanted around the renal artery. Dynamic susceptibility-weighted gradient-echo imaging was used to measure signal-time curves in the renal artery and the renal parenchyma during administration of 1.5 mg/kg BW of an intravascular ultrasmall particle iron oxide (USPIO) contrast agent. From the dynamic series, regional renal blood volume (rRBV), regional renal blood flow (rRBF), and mean transit time (MTT) were calculated. The morphologic degree of stenosis was measured in the steady state using a high resolution 3D contrast-enhanced (CE) MR angiography (MRA) sequence (voxel size = 0.7 x 0.7 x 1 mm(3)). Five patients with renoparenchymal damage due to long standing renal artery stenosis were evaluated. In the animal stenosis model, cortical perfusion remained unchanged for degrees of renal artery stenosis up to 80%. With degrees of stenoses > 80%, cortical perfusion dropped to 151 +/- 54 ml/100 g of tissue per minute as compared to a baseline of 513 +/- 76 ml/100 g/min. In the patients, a substantial difference in the cortical perfusion of more than 200 +/- 40 ml/100 g/min between the normal and the ischemic kidneys was found. The results show that quantitative renal perfusion measurements in combination with 3D-CE-MRA allow the functional significance of a renal artery stenosis to be determined in a single MR exam. Differentiation between renovascular and renoparenchymal disease thus becomes feasible. PMID- 12541250 TI - High-resolution 3D proton spectroscopic imaging of the human brain at 3 T: SNR issues and application for anatomy-matched voxel sizes. AB - In a systematic study on the interdependence of linewidth, signal-to-noise ratio (SNR), and spatial resolution in 3D proton spectroscopic imaging ((1)H-SI) at 3 T, we demonstrate reduced linewidths with increased spatial resolution due to reduced magnetic inhomogeneity within the brain. High-precision quantitative data (0.75-0.094 cm(3)) were obtained for all resolutions, enabling the creation of metabolic maps that display details such as the ventricles, sulci, and gyri. High resolution (1)H-SI allows differences in metabolic ratios to be estimated for anatomically defined regions in gray (GM) and white matter (WM). Seven distinct regions in a healthy brain were anatomically segmented and their metabolic ratios were compared quantitatively. Data from a tumor patient are also presented to demonstrate potential clinical applications. Because of the high resolution, the metabolite ratios could be determined for distinct pathologic regions within the tumor and its surroundings. The method was additionally applied to a patient with patchy Pelizaeus Merzbacher disease (PMD), and compared to single-voxel spectroscopy performed in the same session. High-resolution SI data were demonstrated in our study to allow the direct matching of anatomic and metabolic images. This may enhance the clinical value of (1)H-SI. PMID- 12541251 TI - Improved perfusion-weighted MRI by a novel double inversion with proximal labeling of both tagged and control acquisitions. AB - A novel pulsed arterial spin labeling (PASL) technique for multislice perfusion weighted imaging is proposed that compensates for magnetization transfer (MT) effects without sacrificing tag efficiency, and balances transient magnetic field effects (eddy currents) induced by pulsed field gradients. Improved compensation for MT is demonstrated using a phantom. Improvement in perfusion measurement was compared to other PASL techniques by acquiring perfusion images from 13 healthy volunteers (nine women and four men; age range 29-64 years; mean age 45 +/- 14 years) and second-order image texture analysis. The main improvements with the new method were significantly higher image contrast, higher mean signal intensity, and better signal uniformity across slices. In conclusion, this new PASL method should provide improved accuracy in measuring brain perfusion. PMID- 12541252 TI - In vivo time-resolved quantitative motion mapping of the murine myocardium with phase contrast MRI. AB - Myocardial motion of healthy mice and mice with myocardial infarction was assessed in vivo by phase contrast (PC) cine MRI. The imaging module was a segmented fast low angle shot (FLASH) sequence with velocity compensation in all three gradient directions. To accomplish additional motion encoding, the spin phase was prepared using bipolar gradient pulses, which resulted in a linear dependence between the voxel velocity and spin phase. This method provided accurate quantification of the velocity magnitude and direction of the murine myocardium at a spatial resolution of 234 microm and a temporal resolution of about 10 ms. The acquisition was EKG-gated and the mice were anesthetized by inhalation of 1.5-4.0 vol.% isoflurane at 1.5 l/min oxygen flow. To validate the MRI measurements, an experiment with a calibrated rotating phantom was performed. Deviations between MR velocity measurements and optical assessment by a light detector were lower than 1.6%. During our study, myocardial motion velocities between 0.4 cm/s and 1.7 cm/s were determined for the healthy murine myocardium across the heart cycle. Areas with myocardial infarction were clearly segmented and showed a motion velocity which was significantly reduced. In conclusion, the method is an accurate technique for the assessment of murine myocardial motion in vivo. PMID- 12541253 TI - Rapid measurement of time-averaged blood flow using ungated spiral phase contrast. AB - A novel ungated spiral phase-contrast (USPC) imaging method was developed for rapid measurement of time-averaged blood-flow rates in the presence of pulsatility. The spatial point-spread function was analyzed to provide an intuitive understanding of how spiral trajectories, which sample the k-space origin at every excitation, can mitigate the effects of pulsatility. Pulsatile flow phantom experiments were performed to validate the accuracy and repeatability of the USPC method. The measurement of flow in the renal and femoral arteries of normal volunteers were also performed. The phantom results (error < or = +9%, SD(phantom) < or = 2%, time-averaged pulsatile-flow rates = 3 15 ml/s) and in vivo results (SD(renal) < or = 8%, SD(femoral) < or = 14%) demonstrate the potential of the USPC method for rapidly and repeatedly measuring accurate time-averaged blood flow even in relatively small arteries and in the presence of strong pulsatility. PMID- 12541254 TI - Myocardial tagging with SSFP. AB - This work presents the first implementation of myocardial tagging with refocused steady-state free precession (SSFP) and magnetization preparation. The combination of myocardial tagging (a noninvasive method for quantitative measurement of regional and global cardiac function) with the high tissue signal to-noise ratio (SNR) obtained with SSFP is shown to yield improvements in terms of the myocardium-tag contrast-to-noise ratio (CNR) and tag persistence when compared to the current standard fast gradient-echo (FGRE) tagging protocol. Myocardium-tag CNR and tag persistence were studied using numerical simulations as well as phantom and human experiments. Both quantities were found to decrease with increasing imaging flip angle (alpha) due to an increased tag decay rate and a decrease in myocardial steady-state signal. However, higher alpha yielded better blood-myocardium contrast, indicating that optimal alpha is dependent on the application: higher alpha for better blood-myocardium boundary visualization, and lower alpha for better tag persistence. SSFP tagging provided the same myocardium-tag CNR as FGRE tagging when acquired at four times the bandwidth and better tag- and blood-myocardium CNRs than FGRE tagging when acquired at equal or twice the receiver bandwidth (RBW). The increased acquisition efficiency of SSFP allowed decreases in breath-hold duration, or increases in temporal resolution, as compared to FGRE. PMID- 12541255 TI - Pulsed arterial spin labeling using TurboFLASH with suppression of intravascular signal. AB - Accurate quantification of perfusion with the ADC techniques requires the suppression of the majority of the intravascular signal. This is normally achieved with the use of diffusion gradients. The TurboFLASH sequence with its ultrashort repetition times is not readily amenable to this scheme. This report demonstrates the implementation of a modified TurboFLASH sequence for FAIR imaging. Intravascular suppression is achieved with a modified preparation period that includes a driven equilibrium Fourier transform (DEFT) combination of 90 degrees-180 degrees-90 degrees hard RF pulses subsequent to the inversion delay. These pulses rotate the perfusion-prepared magnetization into the transverse plane where it can experience the suitably placed diffusion gradients before being returned to the longitudinal direction by the second 90 degrees pulse. A value of b = 20-30 s/mm(2) was thereby found to suppress the majority of the intravascular signal. For single-slice perfusion imaging, quantification is only slightly modified. The technique can be readily extended to multislice acquisition if the evolving flow signal after the DEFT preparation is considered. An advantage of the modified preparation scheme is evident in the multislice FAIR images by the preservation of the sign of the magnetization difference. PMID- 12541256 TI - Simulations of thermal tissue coagulation and their value for the planning and monitoring of laser-induced interstitial thermotherapy (LITT). AB - MRI information is widely used for the monitoring of thermal therapies, such as laser-induced interstitial thermotherapy (LITT). However, a detailed knowledge about the relationship between time/temperature exposure and resulting tissue thermal damage is needed to estimate the lesion extent. The aims of this work were to model the thermal response of laser-irradiated tissue and to assess the value of Monte Carlo simulation (MCS) for tumor therapy planning and monitoring. MCS yielded true 3D distributions of laser-induced temperature and thermal damage. These results were compared with 2D MR thermometry data from in vitro experiments performed on animal liver tissue over an ordinary range of laser powers (10-25 W) and irradiation times (5-25 min). Clinical reference data were available from MR-guided liver LITT procedures. MCS correctly predicted the shape of temperature and damage distributions. The quantitative agreement between simulated and measured lesion diameters was within 10% and 15% for the in vitro and in vivo cases, respectively. The simulated 53 degrees C isotherm best estimated the boundary of the final thermal damage (6% variance). This study shows that MCS of interstitial laser coagulation provides unique information that can be of use in the empirical correlation of temperature with thermal damage, and can assist greatly in thermal treatment planning and monitoring. PMID- 12541257 TI - Theoretical and experimental evaluation of detached endcaps for 3 T birdcage coils. AB - The use of detached endcaps for 3 T birdcage coils was investigated both theoretically and experimentally. Finite difference time domain analysis, along with workbench and MRI techniques, were used to map the radiofrequency (RF) B(1) distribution along the coil axis with and without an endcap. Without an endcap the measured B(1) value at the service end of the birdcage was only 45% of the value at the coil's center. This was improved to 85% with a detached endcap of maximum achievable diameter (375 mm), positioned 4 mm from the RF shield. The B(1) field distribution on the patient side of the coil was unaffected by the presence of the endcap. The dependence of the B(1) distribution as a function of endcap diameter was also investigated. Surprisingly, simulations and experiments show that there is an optimum ratio of endcap-to-birdcage coil diameter (approximately 1.08) that gives the best B(1) homogeneity. In the human head the optimized endcap, positioned 16 mm from the RF shield, improves the MRI signal amplitude from 55% to 85% of maximum toward the service end. This novel endcap design is easy to implement with existing birdcage coils, and could prove useful when flexibility in access to the RF coil is required. PMID- 12541258 TI - Visualization of myocardial microstructure using high-resolution T*2 imaging at high magnetic field. AB - The analysis of myocardial microstructure in vivo is important for the determination of myocardial contractility and function. The purpose of the present study was to demonstrate that high-resolution T*2 imaging has the potential to visualize the microstructure of beating, isolated rat hearts. To perform T*2 imaging, a multiple gradient-echo sequence was implemented on an 11.75 Tesla microscopy system. An in-plane resolution of 78 microm and a slice thickness of 250 microm were achieved in 24 min. In comparison to histological sections, the T*2 maps showed an excellent spatial correspondence to the myocardial fiber structure. To demonstrate the utility of this technique, morphologic alterations in myocardial microstructure were investigated in hearts with chronic myocardial infarction. Scar tissue and the extent of the infarcted region were clearly visualized and quantified using high-resolution T*2 imaging. PMID- 12541259 TI - Detection of eye movements from fMRI data. AB - Awake humans make eye movements with amplitudes and frequencies that depend on behavioral state and task. This poses two problems for functional magnetic resonance imaging (fMRI) studies that compare brain activity across tasks. First, motion of the eye in the orbit increases the variance of the MR signal in adjacent regions of the orbitofrontal cortex, hampering activation detection. Second, eye movements are associated with activity in a distributed network of brain areas, confounding comparisons of task activation. Direct measurement of eye movements in the scanner bore is possible with expensive and technically demanding equipment. A method is described that detects eye movements directly from MR data without the use of additional equipment. Changes in the MR time series from the vitreous of the eye were observed that correlated with eye movements, as measured directly with an infrared pupil tracking system. In each of 10 subjects, the variance of the MR time series from the eye vitreous was greater when the subject made eye movements than when the subject fixated centrally (average standard deviation (SD) 99.7 vs. 75.6, P = 0.001). The assessment of eye movements directly from fMRI data may be especially useful for retrospective and meta-analyses. PMID- 12541260 TI - Characterization of aortic root atherosclerosis in ApoE knockout mice: high resolution in vivo and ex vivo MRM with histological correlation. AB - In vivo, cardiac-gated, black-blood, and ex vivo magnetic resonance microscopy (MRM) images of the aortic root, and histopathology data were obtained from 12 transgenic and wild-type (WT) mice. MRM was performed using a black-blood imaging spin-echo sequence with upstream and downstream in-flow saturation pulses to obtain aortic root images in three contrast techniques: proton density-weighted (PDW), T(1)- (T(1)W), and T(2)-weighted (T(2)W). Aortic wall thickness and area were measured and correlated with histopathology data (R > 0.90). Ex vivo lesion components (lipid core, fibrous tissue, and cell tissue) were identified and characterized by differing image contrast in PDW, T(1)W, and T(2)W MRM, and by histopathology. The differences between WT and transgenic mice for maximal wall thickness and area were statistically significant (P < 0.05). This study demonstrates the feasibility of in vivo murine aortic root lesion assessment and ex vivo plaque characterization by MRM. PMID- 12541261 TI - Dynamic MRI reveals that the magnitude of the ischemia-related enhancement in skeletal muscle is age-dependent. AB - This study was designed to evaluate the influence of age on the dynamic contrast enhanced MRI of ischemic tissue. A well-established model of peripheral arterial insufficiency (i.e., the rat hindlimb ischemia after removal of femoral artery) in different age groups (i.e., young, presenescent, and senescent rats) was studied. The analysis of the MR signal demonstrated a marked accumulation of a contrast agent (Gd-DTPA) in the ischemic leg (ischemia-related enhancement, IRE). IRE was an age-related event: 4-month-old rats showed a strong IRE while 12-month old rats and 20-month old rats showed a significantly reduced IRE in comparison to young animals. Histological analysis of the ischemic muscles revealed that there was no evidence of significant necrosis of the muscle tissue but only a weak interstitial fibrosis; CD31-immunostaining revealed a preserved microcirculatory bed. PMID- 12541262 TI - Resolution enhancement in lung 1H imaging using parallel imaging methods. AB - Resolution in (1)H lung imaging is limited mainly by the acquisition time. Today, half-Fourier acquisition single-shot turbo spin-echo (HASTE) sequences, with short echo time (TE) and short interecho spacing (T(inter)) have found increased use in lung imaging. In this study, a HASTE sequence was used in combination with a partially parallel acquisition (PPA) strategy to increase the spatial resolution in single-shot (1)H lung imaging. To investigate the benefits of using a combination of single-shot sequences and PPA, five healthy volunteers were examined. Compared to conventional imaging methods, substantially increased resolution is obtained using the PPA approach. Representative in vivo (1)H lung images acquired with a HASTE sequence in combination with the generalized autocalibrating partially parallel acquisition (GRAPPA) method, up to an acceleration factor of three, are presented. PMID- 12541263 TI - Is TrueFISP a gradient-echo or a spin-echo sequence? AB - It is commonly accepted that TrueFISP (balanced FFE, FIESTA) belongs to the class of gradient-echo (GRE) sequences. GRE sequences are sensitive to dephasing effects of the transverse magnetization between the excitation pulse and echo acquisition, and phase coherence is only established directly after and before excitation pulses. However, an analysis of the phase evolution of transverse magnetization in a TrueFISP experiment shows very close similarities to the echo formation of a spin-echo (SE) experiment. If dephasing between excitation pulses is below +/-pi, TrueFISP exhibits a nearly complete refocusing of transverse magnetization at TE = TR/2. Only signals acquired before and after TR/2 show an additional T*2 sensitivity. PMID- 12541264 TI - Velocity-driven adiabatic fast passage for arterial spin labeling: results from a computer model. AB - Velocity-driven adiabatic fast passage (AFP) is commonly employed for perfusion imaging by continuous arterial spin labeling (CASL). The degree of inversion of protons in blood determines the sensitivity of CASL to perfusion. For this study, a computer model of the modified Bloch equations was developed to establish the optimum conditions for velocity-driven AFP. Natural variations in blood velocity over the course of the cardiac cycle were found to result in significant variations in the degree of inversion. However, the mean degree of inversion was similar to that for blood moving at a constant velocity, equal to the time averaged mean, at peak velocities and heart rates within normal ranges. A train of RF pulses instead of a continuous RF pulse for labeling was found to result in a highly nonlinear dependence of the degree of inversion on RF duty cycle. This may have serious implications for the quantification of perfusion. PMID- 12541265 TI - Cape Asbestos, Barking, health and environment: 1928-1946. AB - BACKGROUND: The mining of chrysotile began in the province of Quebec in the late 1870s. Twenty years later, while annual exports were still measured in hundreds of tons, serious and lethal effects were being reported in England and shortly after in France. In 1913, after a number of years of operating in South Africa, Cape Asbestos Company established a factory in Britain in Barking, Essex. Some 15 years later, in 1929, the local Medical Officer of Health was seeing their asbestotic workers at his Chest Clinic, the local Chest Hospital was investigating them and publishing their findings, and the Coroner began what was to be a long series of inquests. METHODS: Documents from the Institute of Mining and Metallurgy, The Royal School of Mines, Central Library of Imperial College, The Porton and Barking Archives, and documents obtained by legal discovery were utilized in preparing this historical perspective. RESULTS: Despite complaints from the local authority, Parliament was assured that there was no special incidence of disease at Cape, Barking, and that its exhaust ventilation was quite up to the general standard and constantly being improved. Reports on the factory environment and information on the health of Cape's workforce that are available for the years 1928-1934, provide a less sanguine view of the situation, as was confirmed by subsequent health reports on its workforce. CONCLUSIONS: The case of asbestos is considered as a paradigm for the economic considerations that are considered to justify the perpetuation of unsafe and unhealthy occupations, decisions with which medical opinion can be found to concur. PMID- 12541266 TI - Health care utilization of carpenters with substance abuse-related diagnoses. AB - BACKGROUND: Persons in the construction trades in the US have high rates of alcohol and substance abuse. We had the unique opportunity to evaluate health care utilization through private insurance and workers' compensation for a group of carpenters at high risk of injury and substance abuse. METHODS: We identified a cohort of union carpenters. Their claims for medical care through union insurance and workers' compensation, and appropriate measures of time at risk were documented. Using methods of indirect standardization, we compared utilization and costs between carpenters with and without alcohol and substance abuse related diagnoses (ASRD). RESULTS: Through private insurance, those with ASRD had 10% higher outpatient utilization and 2.1 times higher rates of hospitalizations for injury care; 2.6 times higher rates of outpatient care; and 2.9 times higher inpatient admissions for non-injury care. Individuals with ASRD had only modestly increased rates (10%) of outpatient utilization through workers' compensation. CONCLUSIONS: These findings support the need for alcohol and drug abuse prevention and treatment services for this workforce. Operationalizing this among highly mobile workforces, such as the construction trades, is a challenge. While not intending to minimize the problems of alcohol or substance abuse on the job among carpenters, the focus of prevention should not necessarily be in the workplace. PMID- 12541267 TI - Excess of symptoms among radiographers. AB - BACKGROUND: The prevalence and occupational determinants of miscellaneous symptoms referred to as "Darkroom disease" was compared between radiographers and physiotherapists working in British Columbia, Canada. METHODS: The participation rate for the mailout questionnaire survey was 66.1%. A small subset underwent spirometry and methacholine challenge testing. RESULTS: Radiographers had a higher prevalence of most symptoms, with an extreme odds ratio of 11.4 for chemical/metallic taste. The percentage of radiographers with non-specific bronchial hyperresponsiveness (NSBHR) was 3 times higher than that of physiotherapists, although the comparison was not statistically significant. Reporting inadequate ventilation, frequently detecting the odor of X-ray processing chemicals and cleaning up spills within the past 12 months were highly associated with most of the symptoms. CONCLUSIONS: Our results suggest that differences in the prevalence of symptoms represent a complex process, both in exposure and response to the many constituents found in radiographic processing chemicals. Objective testing of health outcomes and more refined exposure measurements are recommended to further investigate occupational health problems of radiographers. PMID- 12541268 TI - The Drake Health Registry Study: findings from fifteen years of continuous bladder cancer screening. AB - BACKGROUND: The Drake Health Registry Study (DHRS) is an ongoing bladder cancer screening program initiated in 1986 due to workers' probable past exposure to the bladder carcinogen, beta-naphthylamine (BNA). METHODS: At periodic screening visits, a health survey is administered and three screening tests are applied to a urine sample, urinalysis (UA), papanicolaou (PAP), and quantitative fluorescence image analysis (QFIA). Positive screens are eligible for a free bladder cystoscopy with random biopsies. RESULTS: Forty of 51 persons eligible for diagnostic evaluation underwent cystoscopy. One person was diagnosed with carcinoma in situ, two with transitional cell papilloma, 14 with dysplasia, two of which developed transitional cell carcinoma; 26 had bladder abnormalities such as chronic inflammation, chronic cystitis, atypical changes, atypia, hyperplasia, or papillary clusters. CONCLUSIONS: The DHRS continues to identify early stage bladder cancer and other abnormalities among workers exposed to BNA before 1981 and generates useful clinical, psycho-social, and epidemiologic data. PMID- 12541269 TI - Occupation and smoking as risk factors for lung cancer: a population-based case control study. AB - BACKGROUND: Lung cancer is associated with occupation, but not much is known about the influence exerted on risk by length of exposure and the joint effect of occupational exposure and tobacco on risk of lung cancer. METHODS: Through a population-based case-control study, we defined risk professions as those that have been associated previously with higher risk of lung cancer. RESULTS: The relative risk seems to increase linearly and significantly with the number of years spent in risk occupations, rising significantly by 28% for every 10 years in a risk profession. Should such occupations be combined with exposure to a smoking habit, then in the case of heavy smokers, a working career spanning 20 years or more in risk occupations would mean tripling the possibility of developing lung cancer from occupation. CONCLUSIONS: The high risks observed indicate a public health problem and indicate that joint exposure to risk professions and tobacco ought to be avoided. We must stress from these results the need for effective education for all workers. PMID- 12541270 TI - An investigation of noise levels in Alberta sawmills. AB - BACKGROUND: Noise exposure in the sawmill industry is an area of concern. This study documents the level of noise exposure in nine sawmills in the province of Alberta, Canada. METHODS: Personal noise monitoring data were collected in nine Alberta sawmills, in winter and in summer (n = 213). Exposures were considered in light of an estimated "real world" noise reduction rating (NRR) calculation assuming use of conventional hearing protection. Limited comparisons were made with spot area monitoring data. RESULTS: Only 10% of the personal monitoring measurements were below the Alberta 8-hr exposure limit of 85 dBA. Twenty-seven percent of the personal monitoring measurements were 95 dBA or higher. Worker enclosures played a large role in reducing noise exposure. There were no significant differences between seasons in noise category distributions (P = 0.61). The planermen and planer infeed operators had the highest percentage of personal monitoring measurements 95 dBA or higher (62% and 82%, respectively). CONCLUSIONS: Based on a conservative formula, a risk of excess noise exposure could exist even when wearing required hearing protection due to very high noise levels found in planing operations in sawmills. PMID- 12541271 TI - Influence of the delta-aminolevulinic acid dehydratase (ALAD) polymorphism on biomarkers of lead exposure in Turkish storage battery manufacturing workers. AB - BACKGROUND: The relationship between delta-aminolevulinic acid dehydratase polymorphism (ALAD) and biomarkers of exposure was investigated in Turkish lead workers in this study. METHODS: Seventy two male lead battery manufacturing workers were selected for the study. Blood lead (BPb) and urinary lead (UPb) concentrations were determined by atomic absorption spectrometry. Erythrocyte ALAD activity and urinary 5-aminolevulinic acid (UALA) were measured spectrophotometrically. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to determine the genotype of the ALAD gene. RESULTS: In total, 51 workers (70.8%) had the ALAD 1-1 genotype, whereas 21 workers (29.2%) had the ALAD 1-2 genotype. No significant relationships were found between the two genotypes and BPb, UPb, and ALAD activity. ALAD1 homozygotes showed significantly higher levels of UALA in comparison with those ALAD2 carriers. CONCLUSIONS: ALAD 1-1 individuals might be an increased risk compared to ALAD2 carriers to disturbance in heme biosynthetic pathway in high lead exposure. PMID- 12541272 TI - Prevalence of hepatitis A virus infection among sewage workers in Georgia. AB - BACKGROUND: Wastewater (WW) workers could have opportunity for direct contact with raw sewage, which might contain hepatitis A virus (HAV). METHODS: A serologic survey of WW workers and a comparison population of roads and drainage workers (RD). Factors potentially associated with anti-HAV positivity were evaluated in univariate and multivariate analyses. RESULTS: Among the 365 WW workers, overall anti-HAV prevalence was 38%, similar to that (35%) of the 166 RD workers (P = 0.5). Prevalence varied by wastewater job type from 45% among the 164 field crew workers to 32% among the 201 treatment plant workers. In multivariate modeling, factors associated with anti-HAV positivity included age > or = 40 years (odds ratio [OR] = 2.4; 95% CI = 1.6-3.7), black compared to other races (OR = 2.4; 95% CI = 1.5-3.8), birth outside the United States (OR = 7.5; 95% CI = 3.0-18.6), a high school education or less (OR 2.1; 95% CI = 1.4-3.2) and work on the field crew compared to RD work (OR 1.6; 95% CI = 1.1-2.4). CONCLUSIONS: These results are consistent with no or a small increased risk of hepatitis A among WW workers, and do not provide a clear mandate for hepatitis A vaccination of these workers. Am. J. Ind. Med. 43: 172-178, 2003. PMID- 12541273 TI - Stressful psychosocial work environment increases risk for back pain among retail material handlers. AB - BACKGROUND: Back pain is a major source of lost work time. Occupational physical activity only accounts for a fraction of low back pain; therefore, there is growing interest in investigating other possible causes of back pain including the psychosocial work environment. METHODS: Material handlers (N = 6,311) in 160 newly opened stores were interviewed at study entry and approximately 6 months later. Factor analysis was used to reduce the 37 psychosocial questionnaire items to seven distinct factors. RESULTS: After adjusting for history of back problems and work-related lifting, risk of back pain was moderately increased among employees who reported high job intensity demands (odds ratio (OR) = 1.8), job dissatisfaction (OR = 1.7), and high job scheduling demands (OR = 1.6). CONCLUSIONS: Modification of the psychosocial work environment for material handlers in large retail stores may help reduce back pain among employees. PMID- 12541274 TI - Quantitative analysis of asbestos burden in women with mesothelioma. AB - BACKGROUND: Lung tissue from 15 women who died from mesothelioma was evaluated for tissue burden of ferruginous bodies and uncoated asbestos fibers. The group contained individuals who had occupational exposure to asbestos and others had family members whose work history included vocations where contact with asbestos containing materials occurred. METHODS: Tissue samples from tumor free lung were digested and filtered and then investigated for ferruginous bodies by light microscopy and asbestos and non-asbestos fibers by analytical transmission electron microscopy (ATEM). Size and type of fibers were also analyzed. RESULTS: Asbestos bodies were found in 13 of the 15 samples and asbestos fibers were found in all cases. The most commonly found uncoated asbestos fiber in these individuals was amosite whereas tremolite was the second most commonly found form. The asbestos fiber burden in these females was often of mixed types. CONCLUSIONS: The asbestos body and fiber burden in these cases show variation in tissue burden. Some cases in this study had appreciable burden, which was attributed to secondhand exposure from occupationally exposed family members. Mesothelioma can occur also in individuals with comparatively low tissue burdens of asbestos. PMID- 12541275 TI - Symptoms and lung function in health care personnel exposed to glutaraldehyde. AB - BACKGROUND: Glutaraldehyde is widely used as a disinfectant for endoscopic equipment. The aim of this study was to investigate work practices and glutaraldehyde exposure in relation to symptoms and lung function. METHODS: A questionnaire was administered to 76 nurses. Exposed nurses (n = 38) also completed lung function tests and visual analogue scales before and after a work session in which glutaraldehyde exposure occurred. Disinfection activities were timed and counted, personal exposures established, and control measures documented. RESULTS: Exposure values above the exposure limit (0.10 ppm) were found for all exposure control methods except for the enclosed washing machine. Skin symptoms were 3.6 times more likely to be reported by exposed workers. None of the other symptoms were significantly associated with glutaraldehyde exposure. There were significant cross-shift reductions in FVC and FEV(1) in the exposed group. No evidence of a dose-response relationship for symptoms or lung function was found. CONCLUSIONS: Further exposure controls for both glutaraldehyde and gloves are required to improve skin care in glutaraldehyde exposed nurses. Exposure monitoring methods also need review. PMID- 12541276 TI - Lifetime principal occupation and risk of Alzheimer's disease in the Kungsholmen project. AB - BACKGROUND: Some studies suggest that manual work is associated with dementia. This study is aimed at identifying the specific occupational categories that may be related to dementia. METHODS: A cohort of 913 non-demented subjects aged 75 + years was longitudinally examined twice over 6 years to detect incident dementia using the DSM-III-R diagnostic criteria. The lifetime longest occupations of all subjects were divided into different categories according to the occupation-based classification system. Data were analyzed with Cox models. RESULTS: During the follow-up period, 260 subjects were diagnosed with dementia (197 with Alzheimer's disease). Manual work was associated with an increased risk of dementia, and the association was dependent on educational level. Compared with non-manual work, manual work involving goods production had a multi-adjusted relative risk (95% CI) of 1.6 (1.0-2.5, P = 0.046) for Alzheimer's disease and 1.4 (0.9-2.1) for dementia. CONCLUSIONS: An association between goods production, manual work and Alzheimer's disease found in this study suggests that factors in the mid twentieth century goods production environment may be involved in the development of Alzheimer's disease. PMID- 12541277 TI - Association between occupational exposure to power frequency electromagnetic fields and amyotrophic lateral sclerosis: a review. AB - BACKGROUND: For the past two decades, there has been concern over electromagnetic exposure and human health. While most research has focused on cancer and reproductive outcomes, there is interest in the relationship between electromagnetic fields (EMF) and neurodegenerative diseases. METHODS: We review epidemiological findings and evidence regarding the association between occupational exposure to power frequency EMFs and amyotrophic lateral sclerosis (ALS). Medline was searched for citations related to occupational hazards and ALS, literature reviews and epidemiological evaluations. RESULTS: Nine out of the ten epidemiological studies that have been conducted on the risk of ALS in relation to occupational exposure to EMF show moderate to strong relative risk estimates that supported a link between them. Although data from these studies was consistent, the causal inference to a link between EMF exposure and ALS is restricted mainly due to the lack of direct information on EMF exposure and incomplete consideration of the other potential risk factors for ALS at workplaces. For instance, electric shock, in particular, is more common in electrical occupations than in any other occupations. CONCLUSIONS: This review concludes that further studies should consider investigating the separate effect of EMF exposure and electrical shocks to make more specific interpretations. On site measurements of EMF should be conducted to include information on EMF exposure from residences as well as workplaces to improve exposure assessment. PMID- 12541278 TI - Occupational issues in episodic care populations. AB - BACKGROUND: Much care for non-emergency problems is provided at "episodic care" settings (e.g., urgent care and emergency room). METHODS: Ninety-four subjects, without known occupational disease, who were seeking care for a chronic or sub acute problem in an episodic care location, were interviewed to assess the frequency and types of occupational health concerns. They were asked whether work probably "caused" or "made worse" their illness and whether their illness "made work difficult." RESULTS: Categorized hierarchically, 20% reported causation, 15% worsening, and 15% interference of illness with work. (Each was categorized in one category only). Only 50% reported absence of a workplace-health interaction. One-third reported that workplace changes could improve their functional status. Nevertheless, workplace factors were discussed only 21% of the time in physician patient encounters. CONCLUSIONS: This study suggests that occupational health issues are frequent in episodic care settings and that the definition of occupational health issues should be broadened to include "made worse" and "illness makes work difficult" as well as the traditional workplace causation definition. Further, episodic care settings present unique opportunities for prevention. PMID- 12541279 TI - National Health Interview Survey mortality among US farmers and pesticide applicators. AB - BACKGROUND: The mortality experience of pesticide-exposed workers across the US has not been thoroughly studied. METHODS: Cox regression mortality analyses adjusted for the complex sample survey design were performed on mortality-linked 1986-1994 National Health Interview Survey (NHIS) data. RESULTS: Nine thousand four hundred seventy-one farmers and pesticide applicators with 571 deaths were compared to 438,228 other US workers with 11,992 deaths. Age-adjusted risk of accidental death, as well as cancers of the nervous and lymphatic/hematopoietic systems, was significantly elevated in male and female pesticide-exposed workers; breast, prostate, and testicular cancer mortality risks were not elevated. CONCLUSIONS: Compared to all other workers, farmers and pesticide applicators were at greater risk of accidental mortality. These pesticide-exposed workers were not at an increased risk of cancers possibly associated with exposure to estrogen analogue compounds, but were at an increased risk of hematopoietic and nervous system cancers. NHIS mortality follow-up represents an important occupational health surveillance instrument. PMID- 12541280 TI - Rapid expression cloning of receptors using epitope-tagged ligands and high-speed cell sorting. AB - BACKGROUND: In this study we describe a new approach for expression cloning of receptors. METHODS: Our approach was based on highly efficient transfer of retroviral cDNA libraries into target cells and detection of receptor-ligand interaction with the use of an antibody directed against an epitope tag on recombinant ligands. Detection of the complex and isolation of receptor transduced cells were achieved by flow cytometry and rare event high-speed cell sorting. Recovery of the cDNA coding for the receptor(s) was achieved by polymerase chain reaction. RESULTS: As a proof-of-concept study we set out to clone the receptor for B-lymphocyte stimulator protein (BlyS), not known at the start of the project but reported while this work was in progress. First, we detected binding of epitope-tagged BlyS to IM9 cells. Second, human T lymphoblasts (CEM cells), which do not bind BlyS, were transduced with a retroviral cDNA library generated from IM9 cells. Transduced CEM cells binding epitope-tagged BlyS protein were identified by flow cytometry. After three sequential rounds of cell sorting, transduced CEM cell populations with high binding capacity for BlyS were identified. To determine the cDNAs conferring binding to the transduced CEM cells, the integrated proviral DNAs were amplified by polymerase chain reaction and analyzed by DNA sequencing. Rescued cDNAs contained Transmembrane Activator and calcium-modulator and cyclophilin ligand (CAML) Interactor (TACI) and B-Cell Maturation factor (BCMA) sequences, representing two published receptors of BlyS. CONCLUSIONS: Our data demonstrated that flow cytometry and high-speed cell sorting combined with transduction of retroviral cDNA libraries and binding of epitope-tagged orphan ligands as a selectable phenotype can be used efficiently for expression cloning of receptors. Of particular interest was our finding that apparently it is not necessary to purify the ligand but that conditioned medium containing the ligand can be used instead. Thus we concluded that our approach shortens the time to identify receptors for many orphan ligands and helps to exploit these receptors as drug targets. PMID- 12541281 TI - Detection of beta-lactamase reporter gene expression by flow cytometry. AB - BACKGROUND: Flow cytometry of gene expression in living cells requires accurate, sensitive, nontoxic fluorescent indicators capable of detecting transcription of specific genes. This is typically achieved by using genes that encode fluorescent proteins or enzymes coupled to promoters of interest. The most commonly used reporters are green fluorescent protein and beta-galactosidase (lacZ). In this study, we characterized the performance of a cell-permeant, ratiometric, beta lactamase substrate, coumarin cephalosporin fluorescein (CCF2/AM). We compared its characteristics with that of the beta-galactosidase/fluorescein di-beta-D galactopyranoside reporter system. METHODS: Jurkat cell lines were generated for beta-lactamase and beta-galactosidase reporters with the use of similar plasmid constructs. Rare event flow cytometric detection for the beta-galactosidase and beta-lactamase reporters were assayed by using mixed populations of negative (WT) and positive (constitutively expressing) cells for each reporter. To determine sensitivity at low reporter copy number, we measured the activity of an unstimulated inducible promoter and detected positive events as a function of substrate incubation time. Technical issues related to data processing and optical configuration are also presented. RESULTS: The low population coefficients of variation afforded by ratiometric detection of the beta-lactamase system improved the statistical performance of the assay in comparison with a single-dye, intensity-based assay, leading to markedly improved detection for low copy number and rare events. At low levels of gene expression, beta-lactamase was detected with approximately 10-fold higher confidence than was beta galactosidase. In rare event detection experiments, cells expressing high levels of beta-lactamase proteins were reliably detected at frequencies of 1:10(6) compared with about 1:10(4) for beta-galactosidase. CONCLUSION: The ratiometric fluorescence readout of the beta-lactamase system based on fluorescence resonance energy transfer allowed more sensitive and accurate detection of gene expression than the currently available beta-galactosidase substrates. Further, the cell permeant nature of the substrate improved experimental convenience. These properties facilitated cell engineering and enabled a variety of applications including selection of rare cells from large populations and measurement of low expressing or downregulated genes. PMID- 12541282 TI - Analysis-only detection of Giardia by combining immunomagnetic separation and two color flow cytometry. AB - BACKGROUND: Giardia is a protozoan parasite of concern to water utilities. Giardia detection relies on cyst isolation and confirmation with the use of fluorescence microscopy. It is of interest to develop a flow cytometric (FCM) method that reliably detects one cyst in 10 L of water. To date all available antibodies have targeted the same epitope on the cyst wall. To achieve a reliable method, two independent probes are required. METHODS: Giardia cysts were spiked into a backwash water sample with and without prior hybridization to peptide nucleic acid (PNA) probes. Immunomagnetic separation (IMS) as a pre-enrichment step was compared with filtration of the water sample. Cysts were recovered with two-color FCM. Those cysts hybridized with PNA and fluorescein isothiocyanate (FITC) were dual stained with monoclonal antibody (mAb) conjugated to phycoerythrin (PE); those not hybridized to PNA were dual stained with mAb-FITC and mAb-PE. RESULTS: A fourfold increase in fluorescent signal intensity was obtained when combining the mAb-PE and PNA probe compared with two-color antibody staining. When combined with IMS, Giardia was successfully identified by FCM, with no false positives detected. CONCLUSIONS: Analysis-only FCM detection of Giardia in water is feasible. Further method development incorporating PNA probe hybridization after IMS is necessary. PMID- 12541283 TI - Automated flow cytometry for acquisition of time-dependent population data. AB - BACKGROUND: The implementation of flow cytometry in many experimental settings can be limited by the extensive amount of sample handling and preparation required for analysis. We describe a system that automatically performs sample handling and flow cytometric analysis, thus allowing one to construct detailed pictures of changes in cell population distributions as a function of time. METHODS: Cell samples from bioreactors were loaded into a microchamber designed to perform all sample preparation steps including washing, fixation, staining, and dilution. The sample was then transported into a sample loop of known volume from which it was injected into the flow cell for determination of cell counts and measurement of scattering and fluorescence parameters. The apparatus was fully automated and controlled with a personal computer equipped with a data acquisition card. An inexpensive mechanism that continuously replenished the sheath fluid was implemented to ensure continuous and uninterrupted operation of the flow cytometer for several days. The device was interfaced with a FACSCalibur equipped with CellQuest software for data acquisition and analysis. RESULTS: The set-up was tested with batch cultures of Saccharomyces cerevisiae expressing the green fluorescent protein (GFP). On-line cell counts showed close agreement with off-line measurements throughout the exponential growth of a yeast culture. The time course of light scattering, GFP fluorescence, and viability distributions provided a detailed description of changes occurring in growing cell cultures based on sampling approximately every 15 min for more than 40 consecutive hours. Therefore, the device could be used to obtain descriptions of the dynamic behavior of cell populations with no user intervention required for several days. CONCLUSIONS: The system significantly expanded the utility of flow cytometry by eliminating cumbersome and time-consuming steps that make the application of flow cytometry impractical in certain situations. It is anticipated that the described set-up will find utility in biotechnology applications such as monitoring of cell cultures, screening of biologically active compounds, and in functional genomics efforts for phenotypic characterizations of cells. PMID- 12541284 TI - Analysis of the distribution of MRI contrast agents in the livers of small animals by means of complementary microscopies. AB - BACKGROUND: Magnetic resonance imaging (MRI) contrast agents contain magnetic molecules such as iron (Fe) or gadolinium (Gd) that are injected in vivo into rats or mice to study their distribution inside the liver. Fluorescent europium (Eu) can be used as a model of Gd to obtain comparable information of this distribution of corresponding contrast agents. In a similar approach, Fe can be attached to Texas Red and used as a model of ferumoxides and be detected by fluorescence. METHODS: To combine and compare the advantages of different microscopic imaging modes, characterization studies were carried out by means of a confocal laser scanning microscope (CLSM), a secondary ion mass spectrometric (SIMS) microscope, and an electron energy loss spectrometric (EELS) microscope. In the case of CLSM, the locations of fluorescent signals inside preparations were determined by factor analysis of biomedical image sequences (FAMIS) and selection of image sequences at emission. RESULTS: By CLSM and FAMIS, we distinguished chelated Eu and Texas Red attached to Fe. By SIMS microscopy, we distinguished Eu and Gd of chlorides and chelates and Fe of a ferumoxide. By EELS microscopy, we distinguished Eu and Gd of chlorides. CONCLUSIONS: Analysis of compounds inside correlative specimens by means of CLSM, SIMS, and EELS microscopes provided complementary results. PMID- 12541285 TI - Flow cytometric quantification of apoptosis and proliferation in mixed lymphocyte culture. AB - BACKGROUND: The one-way mixed lymphocyte culture (MLC) is the classic culture used for studying the allogenic immunoresponse in vitro, but stimulator and responder cell identifications and quantification of apoptotic or proliferative responder cells are unreliable. METHODS: Peripheral blood mononuclear cells were labeled with 5- (and 6-) carboxy fluorescein diacetate succinimidyl ester (CFSE) and stimulated with allogenic unlabeled irradiated cells in unidirectional cultures. Apoptosis was determined by the 7-aminoactinomycin D technique, and the absolute number of each cell population was calculated by adding a fixed number of cells stained with propidium iodide as the reference standard for each test. RESULTS: CFSE labeling of cells under different cultures did not affect the results of proliferation or apoptosis. Data of apoptosis obtained with this method were comparable to those of the monoclonal antibody technique, and the proliferation level determined by [(3)H]-thymidine incorporation or counting the number of proliferative living cells, as proposed in this method, showed a good correlation. CONCLUSIONS: The method presented in this report allows the simultaneous determination of apoptosis and proliferation in MLCs and the analysis of cell phenotype, thereby avoiding the use of radioactivity. This assay opens new perspectives for a better understanding of the mechanisms implied in the establishment or break of tolerance to the graft in solid organ transplants. PMID- 12541286 TI - Quantifying cell scattering: the blob algorithm revisited. AB - BACKGROUND: A method to objectively quantify cell scattering would permit quantitative evaluation of therapies and compounds intended to affect this physiologic process, which has relevance to normal (e.g., development) and pathologic (e.g., metastasis) events. METHODS: A grid-based modified blob analysis was performed on a set of images of Madin-Darby Canine Kidney (MDCK) cells to quantify the following parameters: the number of cellular clusters in each image, the size of the clusters in terms of pixel counts, and the number of cells in each cluster. These parameters were used as measures of cell scattering and were compared with subjective assessments of scattering made by three experienced examiners. RESULTS: The quantitative parameters correlated strongly to subjective assessments. The algorithm displayed a different concept of "clustering" than the examiners and consistently identified more clusters than did the examiners. There was close agreement in the number of cells counted. All three quantitative parameters correlated strongly to the subjective scattering scores, as follows: cluster count (r(s) = -0.765 to -0.789, P < 0.0001), cluster size in pixels (r(s) = 0.838 to 0.845, P < 0.0001), and cluster size in cells (r(s) = 0.758 to 0.804, P < 0.0001). The parameters were continuous, providing greater resolving power than ordinal subjective scores. CONCLUSIONS: The findings confirmed that our algorithm reproduces the traditional classification of scattering with improved resolution, quantification, and objectivity. PMID- 12541287 TI - Nuclear DNA content and genome size of trout and human. PMID- 12541289 TI - B. F. Skinner's technology of behavior in American life: from consumer culture to counterculture. AB - From the 1940s through the 1970s, articles in popular magazines and newspapers presented B. F. Skinner in a wide array of guises, from educational revolutionary and utopian to totalitarian and fascist. Understanding these diverse, and often contradictory, portrayals requires a consideration of the social and political discourses in which they were embedded. In this paper, I suggest that reports of Skinner's work were influenced by a number of cultural categories, from the better living campaign of the 1950s, to the counterculture crusade of the late 1960s. Through this examination, a multifaceted rendering of Skinner's public image that takes into account the nature of his work, the context in which it was produced, and the culture in which it was received is revealed. I propose that the received view of Skinner as maligned behaviorist actually obscures the complexity of his relationship with psychology's public throughout this period. PMID- 12541290 TI - Versemaking and lovemaking--W. B. Yeats' "strange second puberty": Norman Haire and the Steinach rejuvenation operation. AB - This article examines the Steinach rejuvenation operation (a vasectomy) which Norman Haire performed on W. B. Yeats in 1934: while this method is now discredited, many others are still used in similar attempts to "cure" old age and restore youthful vitality. The article deals substantially with ideas which were popular among well-read people and intellectuals in the 1920s and 1930s and discusses whether the poet's late-in-life burst of creativity was caused by a medical breakthrough, Haire's more general medical advice, the placebo effect, or by a combination of factors: questions which still have relevance to theories about age, sexuality, and mind-body relationships. PMID- 12541291 TI - "Automatism" and the emergence of dynamic psychiatry. AB - This article is about the clash of two explanatory paradigms, each attempting to account for the same data of human experience. In the first half of the nineteenth century, physiologists investigated reflex actions and applied a recently coined word, "automatism," to describe actions which, although seeming to arise from higher centers, actually result from automatic reaction to sensory stimuli. Experiments with spinal reflexes led to the investigation of the reflex action of the brain or "cerebral automatisms." Reflex actions of this kind were used to explain everything from acting compulsively to composing symphonies. Physiological explanations of phenomena of this kind seemed insufficient to some and, in the 1880s, Frederic Myers and Pierre Janet developed psychological frameworks for understanding these phenomena, positing hidden centers of intelligence at work in the individual, outside ordinary awareness, which produce what came to be called "psychological automatisms." Their attempts to unify this psychological framework with the existing physiological one failed. Nevertheless, their work played a crucial role in paving the way for what Ellenberger called dynamic psychiatry, which accepts the reality of an unconscious dynamic of the psyche. PMID- 12541292 TI - In the margins of scientific dialogue: Evart Van Dieren contra psychoanalysis. AB - Evart Van Dieren (1861-1940), a Dutch general practitioner who had a passionate aversion against psychoanalysis, wrote two particularly polemical books contra Freud but nevertheless did not succeed in generating much response. In this paper, some of his objections against psychoanalysis are briefly examined and compared with some present day arguments against Freud by Grunbaum and Crews, with the purpose of finding an answer to the question: What does it take to be a successful or unsuccessful critic? PMID- 12541295 TI - Damage to the gills, skin and other tissues by lysenin and the coelomic fluid of the earthworm Eisenia foetida in two teleosts, Tanichthys albonubes and Oreochromis mossambicus. AB - Lysenin is a 33-kDa protein found in the coelomic fluid (CF) of the earthworm Eisenia foetida. Purified lysenin binds specifically to sphingomyelin (SM). In the present studies, we found that the white cloud mountain minnow Tanichthys albonubes and the Mozambique tilapia Oreochromis mossambicus died in solutions of lysenin (at concentrations above 2.5 microg/ml) and CF (0.6%, v/v) within 2 h. The gills of both species of fish were damaged similarly by lysenin and by CF. Most gill lamellae became irregularly bent or curled, with swelling of the epithelial cells of the lamellae. Red blood cells in the lamellar vascular sinuses, in the central venous sinuses, and in the blood vessels of the entire body became swollen and lysed, choking the sinuses. Epithelial cells in the skin were also damaged. When fish of both species were treated with lysenin or CF that had been incubated with SM-liposomes, they did not die. Their behavior remained normal and there was no damage to any cells or tissues. These findings suggest that SM might be involved in the lethal effects of lysenin and CF. It is likely that purified lysenin and lysenin in CF bound to SM in the cell membranes of the tissues mentioned above, damaging the cells. The presence of SM in the gills and skin was confirmed, supporting this hypothesis. The damage to gills and hemolysis might have resulted in lethal respiratory problems. Damage to the skin might disturb the exchange of ions through the skin, hastening death. Damage by lysenin and CF to epithelial cells of the cornea and the wall of the oral cavity was also recognized, but there was no such damage to the intestine. PMID- 12541296 TI - Comparisons of calcium regulation in fish larvae. AB - The purpose of the present study was to compare the ability of larvae of different species, goldfish (Carassius auratus), zebrafish (Danio rerio), and ayu (Plecoglossus altivelis), to regulate their calcium balance. Whole-body Ca(2+) content and Ca(2+) influx in the larvae of the three species, which were incubated in low- (0.02 mM), mid- (0.2 mM), and high- (2.0 mM) Ca(2+) artificial fresh water from embryonic stages, were compared. The Ca(2+) uptake kinetics were determined in zebrafish and goldfish incubated in high- or low-Ca(2+) artificial fresh water. Ca(2+) content of both zebrafish and ayu acclimated to low-Ca(2+) media were significantly lower than those acclimated to mid- or high-Ca(2+) media. However, Ca(2+) contents of goldfish in low-, mid-, and high-Ca(2+) groups showed no significant differences. In goldfish, Ca(2+) influx in the low-Ca(2+) group was significantly higher than those of the mid- and high-Ca(2+) groups. In contrast, the Ca(2+) influx rate in the low-Ca(2+) group was significantly lower than those in the mid- and high-Ca(2+) groups in zebrafish and ayu. Compared to the high-Ca(2+) group, the low-Ca(2+) group of goldfish showed a 13% increase in the maximal velocity (J(max)) and an 84% decrease in the Michaelis constant (K(m)) for Ca(2+) influx. Smaller changes, i.e., an 8% increase in J(max) and a 67% decrease in K(m), were found in zebrafish larvae. Goldfish possess a more effective Ca(2+) regulatory capacity than do zebrafish and ayu. Differences in the strategies for Ca(2+) balance may be associated with different development patterns and environments in which these fish naturally occur. PMID- 12541297 TI - Evidence for a direct link between stress and immunity in the mollusc Haliotis tuberculata. AB - Stress is thought to cause increased disease outbreaks and mortality in a number of invertebrates but currently very little information is available on mechanisms linking physiological states of stress and reduced disease resistance in these organisms. In the present study, we examined the possibility that stress alters immune functions, the principal line of defense against pathogens, in a molluscan model, the abalone Haliotis turbeculata. Immune parameters were investigated in abalones subjected to a 15 min mechanical disturbance which, as indicated by noradrenaline and dopamine hemolymphatic levels, resulted in a transient state of physiological stress. During the application of the stressor, immune parameters such as the number of circulating hemocytes, the migratory activity, the phagocytic capacity and the respiratory burst responses of hemocytes, decreased significantly. All parameters returned to initial values within 15-30 min after the end of the disturbance and a transient period of immunostimulation occurred between 100 and 480 min after the stress for all immune parameters except intracellular superoxide anion production. These results indicate that in the abalone H. tuberculata, as in vertebrates, a link exists between stress and the immune system. This may begin to answer why stress and disease outbreaks are linked in shellfish. PMID- 12541298 TI - Iron uptake by hepatopancreas brush border membrane vesicles (BBMV) of the lobster (Homarus americanus). AB - The uptake of (55)Fe(2+) and solubilized (55)Fe(3+) into brush border membrane vesicles prepared from the hepatopancreas of the Atlantic lobster (Homarus americanus) was investigated. Non-specific surface binding of (55)Fe(2+) at equilibrium to the vesicular surface approximated 57% of total (55)Fe(2+) uptake. (55)Fe(2+) uptake showed temperature sensitivity and was trans-stimulated by a Ca(2+) gradient (at 5mM) directed out. Equilibrated (59)Fe(2+) exchanged for both Cd(2+) and cold Fe(2+). The data obtained in this study are suggestive that at least a portion of ferrous iron absorption may occur by a divalent exchanger mechanism. PMID- 12541299 TI - Distribution and characteristics of D-amino acid and D-aspartate oxidases in fish tissues. AB - The distributions of D-amino acid oxidase (D-AAO, EC 1.4.3.3) and D-aspartate oxidase (D-AspO, EC 1.4.3.1) activities were examined on several tissues of various fish species. Both enzyme activities were commonly high in kidney and liver and low in intestine with some exceptions. After oral administration of D alanine at 5 micromol /g body weight(-1)day(-1) to carp for 30 days, D-AAO activity increased by about 8-, 3-, and 1.5-fold in intestine, hepatopancreas, and kidney, respectively, whereas no increase was found in brain. In contrast, oral administration of D-glutamate or D-aspartate did not show any increase of D AspO activity in any tissues. D-AAO and D-AspO of common carp kidney and hepatopancreas were subcellularly localized in peroxisomes, as clarified in mammals. D-proline was the best substrate for D-AAO in rainbow trout kidney, common carp kidney, and hepatopancreas, followed by D-alanine and D phenylalanine. N-methyl-D-aspartate was the best substrate for D-AspO in rainbow trout kidney and common carp hepatopancreas. The optimal pH for D-AAO in rainbow trout kidney was broad, from 7.4 to 8.2, and that for D-AspO was around 10. D-AAO was inhibited by benzoate known as D-AAO inhibitor and D-AspO was strongly inhibited by meso-tartarate as D-AspO inhibitor. From these results, at least D AAO in fish is considered to work as a metabolizing agent of exogenous and endogenous free D-alanine that is abundant in aquatic invertebrates such as crustaceans and bivalve mollusks, which are potential food sources of these fishes. PMID- 12541300 TI - Molting in the Djungarian hamster (Phodopus sungorus Pallas): seasonal or continuous process? AB - After transfer into a short daylight regimen, the brownish summer pelage of the Djungarian hamster (Phodopus sungorus) changes into the whitish winter phenotype. Although changes in serum prolactin levels are identified as the initiating hormonal signal, morphological data about molting in that species are sparse. The aim of this study was to characterize in detail the summer and winter pelage of the Djungarian hamster and to analyze the alterations in the skin and pelage induced by photoperiodic changes. The main difference between summer and winter hair types is the pattern of pigmentation. In contrast to other mammalian species showing seasonal changes, the winter coat of the Djungarian hamster is not characterized by an increase in hair density. Molting patches were observed at all times, even in the winter coat, showing that the light regimen does not control the process of molting itself but the pattern of pigmentation and eventually the loss of hair during the single molting wave. PMID- 12541302 TI - Prenatal sex ratios influence sexual dimorphism in a reptile. AB - The prenatal environment influences offspring traits in a variety of ways and in a wide range of taxa. For example, maternal allocation of steroids to the eggs influences offspring traits in birds, and in some mammals the intrauterine position influences morphological, behavioural, and physiological traits due to sex-related steroid transfer between sibling fetuses. We show that similar phenomena occur in the common lizard (Lacerta vivipara), a viviparous reptile. Females developing in male-biased clutches had a more masculine allometry (relatively larger heads) at parturition than females developing in female-biased clutches. Males were correspondingly feminized in female-biased clutches. The effects could either be due to diffusion of steroids produced by the offspring or by a general tendency for females to allocate steroids according to the sex ratio of her clutch. Subsequent to parturition, the sexes differed in their growth trajectories depending on sex ratio environment. In males, the difference in allometry between sex ratio environments remained over time, whereas in females the corresponding effect disappeared. PMID- 12541301 TI - Structural and functional aspects of Bufo americanus spermatozoa: effects of inactivation and reactivation. AB - Very little is known about the effects of manipulating toad sperm activity in vitro, and such information is important in the development of a genetic resource bank for bufonid species. The specific objectives of this study were to: 1). identify the optimal inactivation and reactivation solutions for toad spermatozoa collected in urine; 2). establish the length of time toad spermatozoa can be exposed to an inactivation buffer and still resume motility upon reactivation; 3). evaluate the consequence of inactivation on specific sperm characteristics; and 4). characterize the sperm mitochondria vesicle (MV) and its relationship to motility. Reactivated sperm motility was similar after inactivation in either Simplified Amphibian Ringers (SAR) solution or DeBoer's (DB) solution. Diluting the buffer by 80% with water provided the best method for reactivating sperm. Dilutions with NaCl solutions (10-50 mM) produced inferior results. SAR inactivated spermatozoa could remain suspended up to 4 hr and still regain 25% of initial motility upon reactivation in water. Compared to the controls, sperm motility was greater (P<0.01) over time for samples treated with SAR, although forward progression was significantly lower. Furthermore, SAR treatment resulted in sperm samples with a greater number of viable, morphologically normal, and intact MVs over time. Electron microscopy and fluorescent staining confirmed that the toad sperm's MV contains a large number of active mitochondria with very few other cytoplasmic structures. Nearly all spermatozoa exhibiting motility had an intact MV, and dissociation of this structure was clearly related to motility loss. In conclusion, toad spermatozoa can be effectively inactivated and reactivated by varying the osmolality of the external solutions and, although sperm forward progression is reduced, all other characteristics are well maintained. Moreover, the increased number of spermatozoa with intact MV after inactivation suggests the process may help preserve this important structure. PMID- 12541303 TI - Storage of red deer epididymides for four days at 5 degrees C: effects on sperm motility, viability, and morphological integrity. AB - The purpose of this study was to assess the sperm motility, the plasma membrane integrity and the morphology of red deer spermatozoa when maintained within epididymides stored for 4 days at 5 degrees C, and to evaluate whether such stored spermatozoa are able to withstand a refrigeration process. Thirty pairs of testes, with attached epididymides, were collected from 30 hunter-killed mature stags (Cervus elaphus hispanicus), and spermatozoa from each one of the pairs were immediately collected in Triladyl medium, evaluated and refrigerated (Control Group). The remaining testes and epididymides were gradually cooled to 5 degrees C and stored for 1, 2, 3, and 4 days (Experimental Groups), after which spermatozoa were processed as described previously for the control group. The effects on spermatozoa that had been stored within epididymides for various times were determined by assaying sperm motility index (SMI), plasma membrane integrity and sperm morphology (SM). In the same way, SMI and SM were assessed after spermatozoa refrigeration at 5 degrees C for 3 hours in different groups (SMI-R, SM-R). There was no significant decrease in plasma membrane integrity of spermatozoa recovered from epididymides stored at 5 degrees C for 4 days. Similarly, the percentage of morphologically normal spermatozoa remained unaffected during the first 3 days of storage. In contrast, during storage sperm motility evaluation revealed significantly (P<0.05) lower SMI values for samples from epididymides stored 2, 3, and 4 days (47.7+/-3.6, 45.5+/-4.4, 44.1+/-5.2) than that of the control group (57.6+/-1.6). Similar results were obtained after refrigeration of spermatozoa in Triladyl at 5 degrees C. These data suggest that it might be possible to recover functional spermatozoa from red deer epididymides stored at 5 degrees C during several days when epididymal spermatozoa cannot be collected and cryopreserved immediately. PMID- 12541304 TI - Lipids as energy source during salinity acclimation in the euryhaline crab Chasmagnathus granulata dana, 1851 (crustacea-grapsidae). AB - Lipids seem to be the major energy store in crustaceans. Moreover, they are extremely important in maintaining structural and physiological integrity of cellular and sub cellular membranes. During salinity adaptation, energy-demanding mechanisms for hemolymph osmotic and ionic regulation are activated. Thus, the main goal of this work was to verify the possible involvement of lipids as an energy source in the osmotic adaptation process. The estuarine crab Chasmagnathus granulata was captured and acclimated to salt water at 20 per thousand salinity and 20 +/- 2 degrees C, for 30 days. After acclimation, crabs were divided into groups of ten and transferred to fresh water (0 per thousand ), salt water at 40 per thousand salinity, or maintained in salt water at 20 per thousand salinity (control group), without feeding. Before and seven days after the salinity change, wet weight and lipid concentration in gills, muscle, hepatopancreas, and hemolymph were determined according to the colorimetric assay of sulphophosphovanilin. Results show that hepatopancreas lipids were not mobilized during osmotic stress regulation. Gill and muscle lipids were significantly lower in crabs subjected to hypo-osmotic stress than those subjected to the hyper osmotic stress or maintained at the control salinity. Our results point to the occurrence of lipid mobilization and involvement of these compounds in the osmotic acclimation process in C. granulata, but with differences between tissues and the osmotic shock (hypo or hyper) considered. PMID- 12541305 TI - Yolk testosterone varies with sex in eggs of the lizard, Anolis carolinensis. AB - In the green anole (Anolis carolinensis), a lizard with genotypic sex determination, yolk testosterone (T) concentration is greater in male-producing than female-producing eggs at oviposition, but the source and potential effects were not clear from previous studies. If yolk T levels are also sex-specific before eggs are laid, a period during which embryonic steroidogenesis is unlikely, it would strongly suggest that the difference in yolk T is maternally derived. We collected yolk samples from eggs shelling within the oviducts of anesthetized females, and then allowed these females to lay the eggs naturally. Eggs were incubated to hatching to determine sex morphologically, and yolk T concentrations were analyzed by radioimmunoassay. As is the case just after they are laid, yolk T is higher in male than female oviductal eggs. To our knowledge, this is the earliest sex difference reported for any yolk steroid. We suggest that maternally derived yolk T levels could influence sex by differentially affecting male- and female-inducing sperm, because fertilization occurs after yolk deposition and ovulation, while the egg is in the oviduct. Our results, together with those of an increasing number of studies, suggest that a relationship between hormones and vertebrate sex determination may be more widespread than generally appreciated. PMID- 12541306 TI - Partial cloning and expression of the cyclin B gene in the ovary of the bullseye puffer (Sphoeroides annulatus). AB - The bullseye puffer is a marine fish species with great potential for aquaculture in Mexico, and the understanding of its reproductive physiology at every level of biological organization is essential in order to succeed. Several molecules orchestrate the complex process of oocyte maturation and spawning. One of these molecules is cyclin B, which is the regulatory subunit of the maturation promoting factor. In this study, a fragment of the cyclin B gene was isolated from the ovary of the bullseye puffer using an RT-PCR approach. The gene fragment was homologous to the cyclin B2 gene of other vertebrate species. Similar levels of cyclin B gene expression were detected in ovaries at different developmental stages, except for atretic ovaries from captive fish which did not spawn. However, cyclin B gene expression was maintained in captive fish treated with LHRH-a to induce spawning, and appeared to be similar to the pattern observed in wild fish. It is possible that the reduced expression of cyclin B in atretic ovaries is the result of mRNA degradation during atresia. Alternatively, reduced gene expression could be a controlling factor in the process of oocyte reabsorption since cyclin B is required for final oocyte maturation and ovulation. PMID- 12541307 TI - Expression of melanocortin 4 receptor mRNA in the central nervous system of the rat. AB - The melanocortin 4 receptor (MC4-R) plays a pivotal role in maintaining energy homeostasis in rodents and humans. For example, MC4-R deletion or mutation results in obesity, hyperphagia, and insulin resistance. Additionally, subsets of leptin-induced autonomic responses can be blocked by melanocortin receptor antagonism, suggesting that MC4-R-expressing neurons are downstream targets of leptin. However, the critical autonomic control sites expressing MC4-Rs are still unclear. In the present study, we systematically examined the distribution of MC4 R mRNA in the adult rat central nervous system, including the spinal cord, by using in situ hybridization histochemistry (ISHH) with a novel cRNA probe. Autonomic control sites expressing MC4-R mRNA in the hypothalamus included the anteroventral periventricular, ventromedial preoptic, median preoptic, paraventricular, dorsomedial, and arcuate nuclei. The subfornical organ, dorsal hypothalamic, perifornical, and posterior hypothalamic areas were also observed to express MC4-R mRNA. Within extrahypothalamic autonomic control sites, MC4-R specific hybridization was evident in the infralimbic and insular cortices, bed nucleus of the stria terminalis, central nucleus of the amygdala, periaqueductal gray, lateral parabrachial nucleus, nucleus of the solitary tract, dorsal motor nucleus of the vagus (DMV), and intermediolateral nucleus of the spinal cord (IML). By using dual-label ISHH, we confirmed that the cells expressing MC4-R mRNA in the IML and DMV were autonomic preganglionic neurons as cells in both sites coexpressed choline acetyltransferase mRNA. The distribution of MC4-R mRNA is consistent with the proposed roles of central melanocortin systems in feeding and autonomic regulation. PMID- 12541308 TI - Expression of vesicular glutamate transporters, VGluT1 and VGluT2, in axon terminals of nociceptive primary afferent fibers in the superficial layers of the medullary and spinal dorsal horns of the rat. AB - We examined immunohistochemically whether the vesicular glutamate transporters (VGluTs), VGluT1 and VGluT2, might be expressed in synaptic terminals of nociceptive primary afferent fibers within laminae I and II of the medullary and spinal dorsal horns of the rat. VGluT1 immunoreactivity (IR) was intense in the inner part of lamina II but weak in lamina I and the outer part of lamina II. VGluT2-IR was most intense in lamina I and the outer part of lamina II. Expression of VGluTs in synaptic terminals was confirmed by dual immunofluorescence histochemistry for VGluTs and synaptophysin. Expression of VGluTs in axon terminals of primary afferent fibers terminating in laminae I and II was also confirmed immunohistochemically after unilateral dorsal rhizotomy. The dual immunofluorescence histochemistry indicated expression of VGluTs in substance P (SP)-containing axon terminals in lamina I and the outer part of lamina II. Electron microscopy confirmed the coexpression of VGluTs and SP in axon terminals within laminae I and II; VGluTs was associated with round synaptic vesicles at the asymmetric synapses. It was further observed that isolectin IB4, a marker for unmyelinated axons, often bound with VGluT2-immunopositive structures but rarely with VGluT1-immunopositive structures in lamina II. Thus, the results indicated in laminae I and II of the medullary and spinal dorsal horns that both VGluT1 and VGluT2 were expressed in axon terminals of primary afferent fibers, including SP-containing nociceptive fibers and that VGluT in unmyelinated primary afferent fibers terminating in lamina II was primarily VGluT2. PMID- 12541309 TI - Reelin expression during embryonic brain development in Crocodylus niloticus. AB - The expression of reelin mRNA and protein was studied during embryonic brain development in the Nile crocodile Crocodylus niloticus, using in situ hybridization and immunohistochemistry. In the forebrain, reelin was highly expressed in the olfactory bulb, septal nuclei, and subpial neurons in the marginal zone of the cerebral cortex, dorsal ventricular ridge, and basal forebrain. At early stages, reelin mRNA was also detected in subventricular zones. In the diencephalon, the ventral lateral geniculate nuclei and reticular nuclei were strongly positive, with moderate expression in the habenula and focal expression in the hypothalamus. High expression levels were noted in the retina, the tectum, and the external granule cell layer of the cerebellum. In the brainstem, there was a high level of signal in cochleovestibular, sensory trigeminal, and some reticular nuclei. No expression was observed in the cortical plate or Purkinje cells. Comparison with reelin expression during brain development in mammals, birds, turtles, and lizards reveals evolutionarily conserved, homologous features that presumably define the expression profile in stem amniotes. The crocodilian cortex contains subpial reelin-positive cells that are also p73 positive, suggesting that they are homologous to mammalian Cajal Retzius cells, although they express the reelin gene less intensely. Furthermore, the crocodilian cortex does not contain the subcortical reelin-positive cells that are typical of lizards but expresses reelin in subventricular zones at early stages. These observations confirm that reelin is prominently expressed in many structures of the embryonic brain in all amniotes and further emphasize the unique amplification of reelin expression in mammalian Cajal-Retzius cells and its putative role in the evolution of the cerebral cortex. PMID- 12541310 TI - Somatosensory cortex of prosimian Galagos: physiological recording, cytoarchitecture, and corticocortical connections of anterior parietal cortex and cortex of the lateral sulcus. AB - Compared with our growing understanding of the organization of somatosensory cortex in monkeys, little is known about prosimian primates, a major branch of primate evolution that diverged from anthropoid primates some 60 million years ago. Here we describe extensive results obtained from an African prosimian, Galago garnetti. Microelectrodes were used to record from large numbers of cortical sites in order to reveal regions of responsiveness to cutaneous stimuli and patterns of somatotopic organization. Injections of one to several distinguishable tracers were placed at physiologically identified sites in four different cortical areas to label corticortical connections. Both types of results were related to cortical architecture. Three systematic representations of cutaneous receptors were revealed by the microelectrode recordings, S1 proper or area 3b, S2, and the parietal ventral area (PV), as described in monkeys. Strips of cortex rostral (presumptive area 3a) and caudal (presumptive area 1-2) to area 3b responded poorly to tactile stimuli in anesthetized galagos, but connection patterns with area 3b indicated that parallel somatosensory representations exist in both of these regions. Area 3b also interconnected somatotopically with areas S2 and PV. Areas S2 and PV had connections with areas 3a, 3b, 1-2, each other, other regions of the lateral sulcus, motor cortex (M1), cingulate cortex, frontal cortex, orbital cortex, and inferior parietal cortex. Connection patterns and recordings provided evidence for several additional fields in the lateral sulcus, including a retroinsular area (Ri), a parietal rostral area (PR), and a ventral somatosensory area (VS). Galagos appear to have retained an ancestral preprimate arrangement of five basic areas (S1 proper, 3a, 1-2, S2, and PV). Some of the additional areas suggested for lateral parietal cortex may be primate specializations. PMID- 12541311 TI - Critical calpain-dependent ultrastructural alterations underlie the transformation of an axonal segment into a growth cone after axotomy of cultured Aplysia neurons. AB - The transformation of a stable axonal segment into a motile growth cone is a critical step in the regeneration of amputated axons. In earlier studies we found that axotomy of cultured Aplysia neurons leads to a transient and local elevation of the free intracellular Ca2+ concentration, resulting in calpain activation, localized proteolysis of submembranal spectrin, and, eventually, growth cone formation. Moreover, inhibition of calpain by calpeptin prior to axotomy inhibits growth cone formation. Here we investigated the mechanisms by which calpain activation participates in the transformation of an axonal segment into a growth cone. To that end we compared the ultrastructural alterations induced by axotomy performed under control conditions with those caused by axotomy performed in the presence of calpeptin, using cultured Aplysia neurons as a model. We identified the critical calpain-dependent cytoarchitectural alterations that underlie the formation of a growth cone after axotomy. Calpain-dependent processes lead to restructuring of the neurofilaments and microtubules to form an altered cytoskeletal region 50-150 microm proximal to the tip of the transected axon in which vesicles accumulate. The dense pool of vesicles forms in close proximity to a segment of the plasma membrane along which the spectrin membrane skeleton has been proteolyzed by calpain. We suggest that the rearrangement of the cytoskeleton forms a transient cellular compartment that traps transported vesicles and serves as a locus for microtubule polymerization. We propose that this cytoskeletal configuration facilitates the fusion of vesicles with the plasma membrane, promoting the extension of the growth cone's lamellipodium. The growth process is further supported by the radial polymerization of microtubules from the growth cone's center. PMID- 12541312 TI - Toward a unified model of vertebrate taste bud development. PMID- 12541313 TI - Distribution of keratin 8-containing cell clusters in mouse embryonic tongue: evidence for a prepattern for taste bud development. AB - The initiation of the morphogenesis of gustatory papillae is independent of innervation. To address the question of whether taste bud formation is associated with gustatory papilla morphogenesis, we examined developing tongues in mouse embryos from embryonic day 11 to birth. Despite the smooth morphological appearance of the lingual dorsal surface at 13 days of gestation, we observed embryonic taste bud primordia as discrete collections of cytokeratin 8-positive and elongated cells in epithelial placodes in the anterior tongue. In subsequent stages until birth, cytokeratin 8 continues to be expressed in embryonic taste buds distributed in punctuate patterns at regular intervals along rows that are symmetrically located on both sides of the median sulcus in the dorsal anterior developing tongue. Embryonic taste buds were observed in the developing circumvallate papillae from 15.5 days of gestation until birth. The dorsal epithelium of the anterior tongue is not innervated when embryonic taste buds first occur. The increased numbers of embryonic taste buds in developing fungiform papillae until birth are not correlated with the neural invasion of the epithelium. Thus, taste buds occur prenatally more likely independently of the innervation. PMID- 12541314 TI - Neurosecretory identity conferred by the apterous gene: lateral horn leucokinin neurons in Drosophila. AB - The LIM-HD protein Apterous has been shown to regulate expression of the FMRFamide neuropeptide in Drosophila neurons (Benveniste et al. [1998] Development 125:4757-4765). To test whether Apterous has a broader role in controlling neurosecretory identity, we analyzed the expression of several neuropeptides in apterous (ap) mutants. We show that Apterous is necessary for expression of the Leucokinin neuropeptide in a pair of brain neurons located in the lateral horn region of the protocerebrum (LHLK neurons). ap null mutants are depleted of Leucokinin in these cells, whereas hypomorphic mutants show reduced Leucokinin expression. Other Leucokinin-containing neurons are not affected by mutations in ap gene. Co-expression of apterous and Leucokinin is observed exclusively in the LHLK neurons, from larval stages to adulthood. Rescue assays performed in null ap mutants, by expressing Apterous protein under apGAL4 and elavGAL4 drivers, demonstrate the recovery of Leucokinin in the LHLK neurons. These results reinforce the emerging role of the LIM-HD proteins in determining neuronal identity. They also clarify the neuroendocrine phenotype of apterous mutants. PMID- 12541315 TI - Reevaluation of the primary motor cortex connections with the thalamus in primates. AB - Six injections (approximately 1 mm in diameter) of biotinylated dextran amine (BDA) were placed in different locations of the primary motor cortex of the rhesus monkey. Anterograde and retrograde labeling patterns in the thalamus were charted and individual labeled axons traced in continuous serial sections. Both anterograde and retrograde labeling in the thalamus was extensive, spanning several millimeters mediolaterally and including ventral lateral, ventral anterior, centromedian, and centrolateral nuclei. Paracentral, mediodorsal, lateral posterior, and medial pulvinar nuclei were also labeled. Two basic types of corticothalamic axons were identified: small to medium-width, type 1 axons that formed large terminal fields with small boutons, and thick, type 2 axons that formed small terminal fields with large boutons. Within each group, subtypes were identified based on specific features of the axons and terminals: two subtypes of type 1 axons and four subtypes of type 2 axons. The results revealed multiple modes of corticothalamic connectivity: sparsely distributed type 1 axons, dense plexuses of type 1 axons, type 2 axon terminal fields either singly or in clusters, and mixed plexuses of type 1 and type 2 axons. Only some cells in the plexuses were retrogradely labeled; some plexuses did not contain any labeled neurons, and many retrogradely labeled neurons were in the regions devoid of anterograde labeling. These connectivity patterns differed between thalamic nuclei. The results revealed much more complex relationships between M1 and thalamus than were previously thought to exist. It is suggested that this connectivity is neither of exclusively a feedback nature nor perfectly reciprocal but is subserved by a multitude of channels, most likely originating from different populations of cortical neurons, and feeding into a variety of functionally different neuronal networks, with each processing specific information. PMID- 12541316 TI - Cerebrospinal fluid-contacting neurons in the rat spinal cord, a gamma aminobutyric acidergic system expressing the P2X2 subunit of purinergic receptors, PSA-NCAM, and GAP-43 immunoreactivities: light and electron microscopic study. AB - Cerebrospinal fluid-contacting neurons (CSFcNs) occur in various brain regions of lower vertebrates. In mammals, they are restricted to medullospinal areas, and little is known about their projection sites. In the present work, we investigated some morphofunctional characteristics of such neurons in the rat spinal cord by light and electron microscopic immunocytochemistry. CSFcNs expressing the P2X(2) subunit of purinergic receptors were present throughout the spinal cord, though more numerous at lower thoracolumbar and sacral levels. These neurons coexpressed GAD and the polysialylated neural cell adhesion molecule (PSA NCAM), a marker of cellular plasticity. From low thoracic levels downward, tiny amyelinic axons (less than 200 nm in diameter) were tightly packed in bundles, which ran along the ependyma and extended ventrally, eventually concentrating against the walls of the ventral median fissure. In addition to P2X(2), GAD, gamma-aminobutyric acid (GABA), and PSA, these axons expressed GAP-43 immunoreactivity. Moreover, they were labelled along their entire lengths with antibodies against synaptotagmin and synaptophysin, but these failed to reveal intraspinal terminal fields. Taken together, our observations indicate the presence in the rat spinal cord of a highly plastic system of GABAergic CSFcNs that express the P2X(2) subunit of purinergic receptors. The function of this original system remains open to question. In these neurons, the P2X(2) receptors may confer a sensitivity to ATP either present in the CSF or released by nearby neurons of the central autonomic area. PMID- 12541317 TI - Differential cholinergic activation of G proteins in rat and mouse brainstem: relevance for sleep and nociception. AB - Murine models are increasingly used for investigations of sleep, yet no previous studies have characterized cholinergic activation of guanine nucleotide binding proteins (G proteins) in mouse brainstem nuclei known to regulate sleep. This study used in vitro [(35)S]guanylyl-5'-O-(gamma-thio)-triphosphate ([(35)S]GTPgammaS) autoradiography to test the hypothesis that muscarinic cholinergic receptors activate G proteins in C57BL/6J (B6) mouse brainstem. The nuclei studied are homologous to those known in rat and cat to modulate sleep and nociception. In B6 mouse, carbachol significantly increased specific binding of [(35)S]GTPgammaS in the pontine reticular nucleus, caudal part (79%); pontine reticular nucleus, oral part (131%); laterodorsal tegmental nucleus (56%); pedunculopontine tegmental nucleus (86%); dorsal raphe nucleus (53%); dorsal medial periaqueductal gray (54%); and ventrolateral periaqueductal gray (52%) when compared with basal binding. Carbachol-induced G protein activation was concentration-dependent and blocked by atropine, demonstrating mediation by muscarinic receptors. G protein activation by carbachol was heterogeneous across B6 mouse brainstem nuclei. Comparison of [(35)S]GTPgammaS binding between mouse and rat revealed different magnitudes of G protein activation in the pontine reticular formation. In the same pontine reticular formation area of B6 mouse where in vitro treatment with carbachol activates G proteins, in vivo microinjection of cholinomimetics causes a rapid eye movement sleep-like state. These data provide the first direct measurement of muscarinic receptor-activated G proteins in B6 mouse brainstem nuclei known in other species to regulate sleep. PMID- 12541318 TI - Inner S-cone bipolar cells provide all of the central elements for S cones in macaque retina. AB - Synaptic terminals of cones (pedicles) are presynaptic to numerous processes that arise from the dendrites of many types of bipolar cell. One kind of process, a central element, reaches deeply into invaginations of the cone pedicle just below an active zone associated with a synaptic ribbon. By reconstruction from serial electron micrographs, we show that L- and M-cone pedicles in macaque fovea are presynaptic to approximately 20 central elements that arise from two types of inner (invaginating) bipolar cell, midget and diffuse. In contrast, S-cone pedicles, with more synaptic ribbons, active zones/ribbon, and central elements/active zone, are presynaptic to approximately 33 central elements. Moreover, all of these arise from one type of bipolar cell, previously described by others, here termed an inner S-cone bipolar cell. Each provides approximately 16 central elements. Thirty-three is twice 16; correspondingly, these bipolar cells are twice as numerous as S cones. (Specifically, each S cone is presynaptic to four inner S-cone bipolar cells; in turn, each bipolar cell provides central elements to two S cones.) These bipolar cells are presynaptic to an equal number of small-field bistratified ganglion cells, giving cell numbers in 2G:2B:1S ratios. Each ganglion cell receives input from two or more inner S-cone bipolar cells and thereby collects signals from three or more S cones. This convergence, along with chromatic aberration of short-wavelength light, suggests that S-cone contributions to this ganglion cell's coextensive blue-ON/yellow-OFF receptive field are larger than opponent L/M-cone contributions via outer diffuse bipolar cells and that opponent L/M-cone signals are conveyed mainly by inner S-cone bipolar cells. PMID- 12541319 TI - Fiber connections of the torus longitudinalis in a teleost: Cyprinus carpio re examined. AB - Fiber connections of the carp torus longitudinalis were re-examined by means of tract-tracing methods. The torus longitudinalis projected mainly to the stratum marginale of the optic tectum, area pretectalis, and corpus cerebelli. The stratum marginale was anterogradely labeled only by biocytin, but not by horseradish peroxidase. Because the stratum is composed of extremely fine axons of the small toral neurons, this may be ascribed to different molecular weights of the tracers. The main afferent sources to the torus longitudinalis were the nucleus subvalvularis, which was located beneath the nucleus lateralis valvulae, the nucleus subeminentialis pars magnocellularis, and neurons along the posterior mesencephalo-cerebellar tract. Some labeled cells also appeared in the area pretectalis, nucleus paracommissuralis, optic tectum, and torus semicircularis. In a previous paper, it was incorrectly reported that the valvula cerebelli was the main source of afferents to the torus longitudinalis. Here we report the reason for the previous mistake in relation to the techniques employed. PMID- 12541320 TI - Chemokine receptors in the brain: a developing story. PMID- 12541321 TI - Developmental changes in the expression of chemokine receptor CCR1 in the rat cerebellum. AB - Chemokines are small, soluble proteins that regulate leukocyte migration, adhesion, and proliferation. Recent evidence suggests that chemokine receptors are expressed in the central nervous system and that their functions extend beyond their roles in inflammation. Specific chemokines and their receptors are implicated in cerebellar development. In this study, we evaluated the expression of beta-chemokine receptor CCR1 in the immature and adult rat cerebellum and report striking developmental changes in CCR1 expression. Reverse transcriptase polymerase chain reaction assays of cerebellum revealed moderate increases in CCR1 mRNA expression from postnatal day (P) 3 to adulthood. Light and confocal microscopy were used to evaluate developmental changes in the neuroanatomical and cell-specific distribution of CCR1 immunoreactivity. CCR1 immunoreactivity was detected as early as P3 and peaked between P7 and P21. The predominant CCR1 immunoreactive neuronal cell types included granule cells of the internal granular layer, Purkinje cells, Golgi cells, and molecular layer interneurons; Bergmann glia, astrocytes, and resting microglia also expressed CCR1. In contrast, granule cells in the external germinal layer, descending granule cells, and activated microglia rarely expressed CCR1. We also evaluated the expression of the CCR1 ligand macrophage inflammatory protein-1alpha (MIP-1alpha/CCL3). Two cell populations expressed MIP-1alpha: physiologically activated microglia in white matter (P7-P14) and Purkinje cells (P7-adult). MIP-1alpha-positive cells were frequently located near the processes and cell bodies of CCR1-immunoreactive cells, during times of neuronal and glial maturation (second and third postnatal weeks). These findings provide support for the hypothesis that CCR1 plays a role in postnatal cerebellar development. PMID- 12541322 TI - Epidermal reinnervation after intracutaneous axotomy in man. AB - Two distinct patterns of reinnervation occur after injury to the cutaneous nerves: regenerative growth of the injured nerve and "collateral sprouting" of neighboring intact nerves. We describe two complementary models of regrowth of transected small sensory fibers in human skin. The "incision" model uses a circular incision that transects the subepidermal plexus, resulting in Wallerian degeneration of the nerve fibers that enter the incised cylinder, leaving a defined zone of denervated dermis and epidermis. The "excision" model utilizes an identical incision, followed by removal of the incised cylinder of skin, leaving a denervated area in which Schwann cells are absent. In the incision model, the earliest reinervation of denervated epidermis occurred by collateral sprouting from the terminals of epidermal axons from just outside the incision line. These axon terminals extended horizontally across the incision line and through the superficial layers of the epidermis, beneath the stratum corneum. By 13 days, numerous regenerating axons appeared in the deeper dermis derived from transected axons. These regenerating axons grew toward and ultimately into the epidermis, so that epidermal axonal density had normalized by 30-75 days. The invasion of these axons was associated with regression of the horizontally growing collateral sprouts. In the excision model, new fibers arose by terminal elongation of the epidermal axons outside the incision line, as in the incision model, and especially by collateral branching of epidermal fibers at the incision margins. These collaterals reached the epidermal surface of the basal lamina at the dermal epidermal junction and then grew slowly toward the center of the denervated circle. In contrast to the incision model, however, complete reinnervation was not achieved even after 23 months. These models can be used to study reinnervation of denervated skin in man in different injury models and have relevance for exploring the stimuli for axonal growth and remodeling. PMID- 12541323 TI - Tectothalamic visual projections in turtles: their cells of origin revealed by tracing methods. AB - In two species of turtle (Emys orbicularis and Testudo horsfieldi), retrograde and anterograde tracer techniques were used to study projections from the optic tectum to the nucleus rotundus (Rot) and to the dorsal lateral geniculate nucleus (GLd). The ipsilateral Rot received the most massive tectal projections, stemming from numerous neurons located in the stratum griseum centrale (SGC). These neurons varied in size and shape, many of them having a wide zone of dendritic arborization within both the (SGC) and the stratum griseum et fibrosum superficiale (SGFS). Projections from the tectum to the GLd were ipsilateral, were extremely scarce, and arose from a small number of neurons of various shapes situated in the SGFS; these cells were, as a rule, smaller than those projecting to the Rot. For the most part, these neurons were radially oriented, with rather restricted dendritic arborizations in the most superficial sublayers of the SGFS; smaller numbers of projection neurons were horizontally oriented, with long dendrites branching throughout the layer. Some neurons located in the stratum griseum periventriculare (SGP) projected to both the Rot and the GLd. Most of these neurons had dendritic arborizations within the retinorecipient zone of the SGFS. We were unable to rule out the possibility that some cells projecting to the GLd were situated in the SGC. Both the GLd and the main body of the Rot did not contain neurons projecting to the optic tectum. Thalamic neurons projecting to the tectum were observed in the ventral lateral geniculate nucleus, the intergeniculate leaflet and the interstitial nuclei of the tectothalamic tract, and the nucleus of the decussatio supraoptica ventralis. The question of whether variation in the laminar organization of the tectorotundal and tectogeniculate projection neurons in reptiles, birds, and mammals may be related to different degrees of differentiation of the tectal layers is discussed. PMID- 12541324 TI - Identification and localization of a neprilysin-like activity that degrades tachykinin-related peptides in the brain of the cockroach, Leucophaea maderae, and locust, Locusta migratoria. AB - Neprilysin (NEP) is an endopeptidase, which has an important role in the inactivation of mammalian tachykinins. NEP-like activity has also been found in the brain of several insects; however, the lack of information about the cellular localization of this peptidase has hindered our understanding of its role in peptidergic signaling in insects. We now provide evidence that membrane-bound NEP is involved in the inactivation of tachykinin-related peptides in the brain of the cockroach, Leucophaea maderae, and the locust, Locusta migratoria. The L. maderae enzyme cleaved the cockroach peptide LemTRP-1 and the mammalian NEP substrate [DAla(2),Leu(5)]enkephalin at the Gly-Phe peptide bond. The enzyme was acted upon by the NEP inhibitors phosphoramidon (IC(50), 0.64 microM) and thiorphan (IC(50), 1.23 microM), and the detergent-solubilized enzyme had an Mr of approximately 300,000 and a neutral pH optimum. This endopeptidase cleaved another insect tachykinin-related peptide, CavTK-II, in a predictable manner at the Ala-Phe peptide bond, suggesting that the peptidase can hydrolyse tachykinin related peptides with different structures. NEP activity was histochemically localized in several, but not all, regions of neuropil in the brain of L. maderae, including the central body, the lobula of the optic lobe, and the tritocerebrum. All of these regions are known to receive neuronal processes containing tachykinin-related peptides. A slightly different distribution pattern for NEP was observed in the brain of L. migratoria. Again, NEP was localized to regions of the neuropil that also display tachykinin-related peptide immunoreactivity. The data reported provide evidence for an evolutionary conserved role for NEP in the inactivation of tachykinin-related peptides in the brain. PMID- 12541325 TI - Progressive recovery of learning during regeneration of a single synapse in the medicinal leech. AB - The leech escape reflex-shortening of the body-can change with nonassociative conditioning, including sensitization, habituation, and dishabituation. Capacity for sensitization, which is an enhancement of the reflex, is lost when a single S interneuron is ablated, but the reflex response itself remains. In the present experiments, the S-interneuron's axon in the living leech was filled with 6 carboxyfluorescein (6-CF) dye and cut with an argon laser microbeam (lambda = 488 nm). In contrast to sham-operated animals, axotomized preparations did not sensitize, reflecting the key role of the S-cell. By 2 weeks or more, S-cell axons had regenerated and reestablished synapses at their usual locations with neighboring S-cells. By 4 weeks, this restored the ability to sensitize to a level indistinguishable from that of controls, but an intermediate state of recovery was seen from 2-3 weeks after injury-a period not previously examined. The small capacity for sensitization among newly regenerated preparations was significantly lower than in sham controls but appeared higher than in animals whose cut S-cell axon had not regenerated its synapse. The results confirm the crucial role of the S-cell in sensitization. Moreover, full sensitization does not occur immediately upon synapse regeneration. PMID- 12541326 TI - Changes in mouse barrel synapses consequent to sensory deprivation from birth. AB - Neonatal sensory deprivation induced by whisker trimming affects significantly the functional organization of receptive fields in adult barrel cortex. In this study, the effects of deprivation on thalamocortical synapses and on asymmetrical and symmetrical synapses not of thalamic origin were examined. Thalamocortical synapses were labeled by lesion-induced degeneration in adult (postnatal day 60) mice subjected to whisker trimming from birth, other synaptic types were unlabeled. Brains were processed for electron microscopy, and numerical densities of synapses were evaluated by using stereologic approaches for whisker trimmed vs. control animals. Results demonstrated no change in nonthalamic, asymmetrical synapses; however, a decrease of 52% in the numerical density of symmetrical synapses (46.3 vs. 88.5 million per mm(3); Z = -2.121; P < 0.05) and a decrease of 43% in the numerical density of thalamocortical synapses (57.5 vs. 102.33 million per mm(3); Z = -2.121; P < 0.05) were observed after deprivation. Thus, experience-dependent plasticity of receptive fields in barrel cortex involves directly axons of both extrinsic and intracortical origin. The proportion of thalamocortical axospinous to axodendritic synapses was the same in control vs. deprived animals: in each instance, 80% of the synapses were axospinous (Z = 0.85; P = 0.2). These results suggest that neither excitatory neurons, whose thalamocortical synapses are primarily axospinous, nor inhibitory neurons, whose thalamocortical synapses are mainly axodendritic (White [1989] Cortical Circuits. Synaptic Organization of the Cerebral Cortex; Structure, Function, and Theory. 1989; Boston: Birkhauser), are affected preferentially by the deprivation associated decrease in thalamocortical synapses. PMID- 12541327 TI - The relationship between hodological and cytoarchitectonic organization in the vestibular complex of the 11-day chicken embryo. AB - To understand the relationship between structure and function in specific brain regions, it is necessary to ascertain which anatomical features are physiologically relevant. Physiological studies of brain function traditionally have been set in the context of anatomical features based on cytoarchitectonics and myeloarchitectonics, but the relationship between structure and function in this context can be complex. Alternative schemes of anatomical organization, such as that based on hodology (the mapping of projections) may provide greater insight. Here, we make a direct comparison of the hodological and the cytoarchitectonic organization of the vestibular complex in the mid-term chicken embryo, using retrograde tracing and three-dimensional reconstruction. In one set of experiments, vestibulospinal and vestibulo-ocular neuron groups were selectively labeled with biotin dextran-amines and aligned with the cytoarchitectonically defined vestibular nuclei in alternating sections that were then combined into intercalated three-dimensional models. This allowed a semiquantitative analysis of the apportionment of individual hodological groups among cytoarchitectonic nuclei. In another set of experiments, vestibulospinal and vestibulo-ocular neuron groups were labeled differentially with fluorescent dextran-amines, three-dimensionally reconstructed, and subjected to a quantitative analysis of spatial overlap. Our results provide the first three dimensional representation and quantitative analysis of the hodological compartmentalization of the vestibular complex (the "hodological mosaic"). They also show directly how each hodologically defined neuron group relates to the conventional vestibular nuclei, underscoring the fact that the units of the hodological mosaic do not bear a one-to-one correspondence to the cytoarchitectonic nuclear divisions. Some hodologically defined groups are localized to restricted portions of a nucleus, whereas others overlap multiple nuclei. Thus, hodology and cytoarchitectonic features appear to be separately regulated in the vestibular complex of the chicken embryo, possibly through different sets of positional specification mechanisms. The three-dimensional representations we present here provide a foundation for integrating anatomical, physiological, developmental, and evolutionary studies of the vestibular system. PMID- 12541329 TI - Native American mtDNA prehistory in the American Southwest. AB - This study examines the mtDNA diversity of the proposed descendants of the multiethnic Hohokam and Anasazi cultural traditions, as well as Uto-Aztecan and Southern-Athapaskan groups, to investigate hypothesized migrations associated with the Southwest region. The mtDNA haplogroups of 117 Native Americans from southwestern North America were determined. The hypervariable segment I (HVSI) portion of the control region of 53 of these individuals was sequenced, and the within-haplogroup diversity of 18 Native American populations from North, Central, and South America was analyzed. Within North America, populations in the West contain higher amounts of diversity than in other regions, probably due to a population expansion and high levels of gene flow among subpopulations in this region throughout prehistory. The distribution of haplogroups in the Southwest is structured more by archaeological tradition than by language. Yumans and Pimans exhibit substantially greater genetic diversity than the Jemez and Zuni, probably due to admixture and genetic isolation, respectively. We find no evidence of a movement of mtDNA lineages northward into the Southwest from Central Mexico, which, in combination with evidence from nuclear markers, suggests that the spread of Uto-Aztecan was facilitated by predominantly male migration. Southern Athapaskans probably experienced a bottleneck followed by extensive admixture during the migration to their current homeland in the Southwest. PMID- 12541330 TI - Genetic diversity of JC virus in the modern Filipino population: implications for the peopling of the Philippines. AB - The Philippines is generally believed to have been established by various peoples who migrated from neighboring areas. To gain new insights into the peopling of the Philippines, we used the JC virus (JCV) genotyping approach. We collected about 50 urine samples on each of two representative islands of the Philippines, Luzon and Cebu. DNA was extracted from the urine samples and used to amplify the 610-bp region (IG region) of the viral genome. For each island, we determined about 20 IG sequences, from which a neighbor-joining phylogenetic tree was constructed to classify the JCV isolates detected into distinct genotypes. The predominant genotype detected was SC, the Southeast Asian genotype. Minor JCV genotypes were SC/Phi, B1-a, and B3. SC/Phi was a subcluster of SC and has not been detected in areas other than the Philippines. B1-a was detected previously in mainland China, Pamalican Island (Palawan, Philippines), and Taiwan (an aboriginal tribe). B3 was classified in this study into two subgroups, one (B3-a) containing three Luzon isolates and several Chinese, Thai, and Uzbek isolates, the other (B3-b) containing two Luzon, one Cebu, and one Indonesian isolate. These findings suggest that the modern Filipino population was formed not only by Southeast Asians carrying SC but also by a few distinct ethnic groups carrying SC/Phi, B1-a, and B3-a or -b. PMID- 12541331 TI - Two probable cases of treponemal disease of Medieval date from England. AB - Two British Medieval skeletons are described. Paleopathological examination suggests that treponemal disease was present in each case. Radiocarbon dating indicates that one is firmly pre-Columbian, and the other of likely pre-Columbian date. The implications for our understanding of the history and geographic spread of the treponemal diseases, particularly venereal syphilis, are discussed. PMID- 12541333 TI - Comparative functional analysis of skull morphology of tree-gouging primates. AB - Many primates habitually feed on tree exudates such as gums and saps. Among these exudate feeders, Cebuella pygmaea, Callithrix spp., Phaner furcifer, and most likely Euoticus elegantulus elicit exudate flow by biting into trees with their anterior dentition. We define this behavior as gouging. Beyond the recent publication by Dumont ([1997] Am J Phys Anthropol 102:187-202), there have been few attempts to address whether any aspect of skull form in gouging primates relates to this specialized feeding behavior. However, many researchers have proposed that tree gouging results in larger bite force, larger internal skull loads, and larger jaw gapes in comparison to other chewing and biting behaviors. If true, then we might expect primate gougers to exhibit skull modifications that provide increased abilities to produce bite forces at the incisors, withstand loads in the skull, and/or generate large gapes for gouging. We develop 13 morphological predictions based on the expectation that gouging involves relatively large jaw forces and/or jaw gapes. We compare skull shapes for P. furcifer to five cheirogaleid taxa, E. elegantulus to six galagid species, and C. jacchus to two tamarin species, so as to assess whether gouging primates exhibit these predicted morphological shapes. Our results show little morphological evidence for increased force-production or load-resistance abilities in the skulls of these gouging primates. Conversely, these gougers tend to have skull shapes that are advantageous for creating large gapes. For example, all three gouging species have significantly lower condylar heights relative to the toothrow at a given mandibular length in comparison with closely related, nongouging taxa. Lowering the height of the condyle relative to the mandibular toothrow should reduce the stretching of the masseters and medial pterygoids during jaw opening, as well as position the mandibular incisors more anteriorly at wide jaw gapes. In other words, the lower incisors will follow a more vertical trajectory during both jaw opening and closing. We predict, based on these findings, that tree-gouging primates do not generate unusually large forces, but that they do use relatively large gapes during gouging. Of course, in vivo data on jaw forces and jaw gapes are required to reliably assess skull functions during gouging. PMID- 12541332 TI - Widespread occurrence of Mycobacterium tuberculosis DNA from 18th-19th century Hungarians. AB - A large number (265) of burials from 1731-1838 were discovered in sealed crypts of the Dominican Church, Vac, Hungary in 1994. Many bodies were naturally mummified, so that both soft tissues and bones were available. Contemporary archives enabled the determination of age at death, and the identification of family groups. In some cases, symptoms before death were described and, occasionally, occupation. Initial radiological examination of a small number of individuals had indicated calcified lung lesions and demonstrable acid-fast bacteria suggestive of tuberculosis infection. Tuberculosis was endemic in 18th 19th century Europe, so human remains should contain detectable Mycobacterium tuberculosis complex (MTB) DNA, enabling comparisons with modern isolates. Therefore, a comprehensive examination of 168 individuals for the presence of MTB DNA was undertaken. Specific DNA amplification methods for MTB showed that 55% of individuals were positive and that the incidence varied according to age at death and sampling site in the body. Radiographs were obtained from 27 individuals and revealed an association between gross pathology and the presence of MTB DNA. There was an inverse relationship between PCR positivity and MTB target sequence size. In some cases, the preservation of MTB DNA was excellent, and several target gene sequences could be detected from the same sample. This information, combined with MTB DNA sequencing data and molecular typing techniques, will enable us to study the past epidemiology of TB infection, and extends the timeframe for studying changes in molecular fingerprints. PMID- 12541334 TI - Dietary constraints on encephalization in primates. AB - Encephalization, and its relationship to potential selective forces, have been a focus of many studies of primate adaptation. It has been argued that gut size may constrain brain mass because these two types of "expensive tissue" (among others) compete in their metabolic requirements (Aiello and Wheeler [1995] Curr. Anthropol. 36:199-221). Following from the inverse correlation of gut size with diet quality, the expensive tissue hypothesis predicts that differences in diet quality are positively correlated with differences in brain mass, once the correlation of each variable with body mass is taken into account. We tested this prediction using both nonphylogenetic and phylogenetic methods. The results of both methods are consistent with predictions made by the expensive tissue hypothesis. We also discuss several examples of independent contrasts that are consistent with the hypothesis (e.g., Colobinae vs. Cercopithecinae), as well as some that are not (e.g., Tarsius vs. anthropoidea). Overall, the results indicate that improved diet quality, by allowing reduction in relative gut mass, is one mechanism involved in increased encephalization. PMID- 12541335 TI - Demographic and life-history patterns in a population of ring-tailed lemurs (Lemur catta) at Beza Mahafaly Reserve, Madagascar: a 15-year perspective. AB - Over 15 field seasons (1987-2001), we collected census and life-history data on a population of individually identified ring-tailed lemurs at the Beza Mahafaly Reserve, Madagascar. No significant difference was found in population size over the study period, though a marked decline in the population occurred following a 2-year drought. The population rebounded rapidly after the immediate postdrought period. There was nearly a complete replacement of individuals over the study period. Average group size is 11.5 animals, and adult male to female sex ratio is 0.92. Most females reproduce annually, and the average fecundity rate is 84.3%. The greatest variability in fecundity is found among old females. We suggest that ring-tailed lemur females follow an "income breeding" strategy, i.e., females use maximum resources during reproduction rather than relying on fat stores, as do "capital breeders." Infant mortality to 1 year of age in a nondrought year is 52%, higher than infant mortality in small to medium-sized anthropoids. The oldest known female was 18 years old in 2001. We suggest that 18-20 years may represent the maximum life-span for wild ring-tailed lemurs. Because males regularly emigrate from the population, we have no data regarding male life-span; however, there is some indication that males do not survive as long as females. Group fission has occurred three times: twice from one parent group living in the driest area of the reserve, with the most dispersed food resources. We suggest that the reproductive strategy that has evolved in this species, wherein females reproduce early in life and annually until old age, is a response to the unusual climate and environmental conditions under which Lemur catta has evolved. PMID- 12541336 TI - Brief communication: coexistence of two distinct patterns in the surname structure of Sicily. AB - The extent of variation in the migratory movements that occurred in Sicily was evaluated using surname data taken from the telephone directories of the 390 communes of the island. The surname distribution of each commune was linearized by a log-log transformation, and a significant fit to a linear regression model was found in almost all cases. Interestingly, the slope of the regression line appeared to be a sensitive indicator of the different level of isolation associated with each Sicilian commune. By this approach, two distinct groups of communes, showing a higher or lower degree of isolation, were obtained, and two independent analyses of the surname structure of Sicily were carried out. A first multidimensional scaling analysis, based on the more isolated communes, yielded evidence for a more ancient pattern, characterized by a geographical gradient along the east-west axis. The same analysis, addressed to the less isolated communes, instead highlighted a wide network of interactions between geographically distant zones of the island. The fitting of the surname distribution to the log-log model allowed for the detection of a narrow subset of 35 Sicilian communes, whose significantly higher degree of isolation was statistically proved by the parallelism test. We believe that a genetic analysis focused on such specific zones of the island could reveal ancient patterns of differentiation, thus helping to answer the controversial question of the genetic history of Sicily. PMID- 12541337 TI - Pathological alterations in the archaic Homo sapiens cranium from Eliye Springs, Kenya. AB - This paper reports on the results of a first computerized tomography (CT)-based study of the Middle Pleistocene matrix-filled skull KNM-ES 11693 from Eliye Springs at Lake Turkana. Ectocranially, the hominid cranium exhibits a remarkable enlargement of the vault symmetrical to the sagittal suture and a porotic surface covering most of the vault. CT analysis further revealed a strong thickening of the cranial vault as well as other relevant aspects. Differential diagnosis suggests that the changes of the Eliye Springs cranium were probably caused by chronic anemia in the childhood or youth of this individual. PMID- 12541339 TI - [Long-term results of surgical treatment of intractable Meniere's disease for control of vertigo]. AB - OBJECTIVE: To acquire the long-term (an average of over 10 years follow-up) results of surgical treatment of patients with Meniere's disease for control of vertigo. METHOD: Endolymphatic sac surgery was done in 6 cases (1 case of endolymphatic sac decompression and 5 cases of sac-mastoid shunting) and translabyrinth vestibular neurotomy was operated on 6 cases. RESULT: Complete relief from symptoms of vertigo occurred in the 6 cases through vestibular neurotomy and there were no recurrence in 11-13 years follow-up. Three of 6 cases with endolymphatic sac surgery had complete control of vertigo and were cured clinically. Symptoms of vertigo of the rest 3 cases were effectively controlled. CONCLUSION: Endolymphatic sac surgery and translabyrinth vestibular neurotomy are effective way for control of vertigo of patients with intractable Meniere's disease. Translabyrinth vestibular neurotomy are more effective for control of vertigo for appropriately selected patients. PMID- 12541340 TI - [The high-resolution CT scan of facial nerve injury following temporal bone fracture]. AB - OBJECTIVE: To discuss the applied value of the high resolution CT scan in the facial nerve injury induced by the temporal bone fracture. METHOD: Thin slice temporal bone CT scans were conducted in 28 cases of facial nerve injuries following head trauma and operations were performed according to temporal bone CT scan and clinical manifestation. RESULT: 6 cases recovered nearly to normal; 18 cases partially recovered and 4 cases were without any change. CONCLUSION: The high resolution CT scan can show the fracture line's location and trend. It is of help to decide the injury site as well as other related injuries and to choose an appropriate surgical approach. PMID- 12541341 TI - [Effect of decompression or transplantation of the facial nerve on serious advanced facial paralysis]. AB - OBJECTIVE: To investigate operative effect on serious advanced facial paralysis. METHOD: 28 cases (28 sides) with serious facial paralysis underwent decompression or transplantation of facial nerve. RESULT: After following up 0.5-2 years postoperatively, the total, most or partial and no recovery were 14 cases (50%), 12 cases (42%) and 2 cases (7.1%) respectively. CONCLUSION: The operation performed as early as possible can be an effective procedure for correcting serious advanced facial paralysis if conservative treatment could be confirmed to be helpless. PMID- 12541342 TI - [Malignant external otitis: 4 cases report]. AB - OBJECTIVE: To improve the level of diagnosis and treatment of malignant external otitis (MEO). METHOD: Four cases with MEO were reported. RESULT: Three in four cases were followed up. One patient was cured, and the others died of intracranial infection. CONCLUSION: Early diagnosis of MEO is very important for treatment. The patients of MEO with diabetes often died of intracranial infection. PMID- 12541343 TI - [Analysis of measured results of transient-evoked otoacoustic emissions]. AB - OBJECTIVE: In order to provide grounds for clinical diagnosis, the normal sensitive parameters of the transient evoked otoacoustic emissions (TEOAE) in normal ears were studied. METHOD: 60 ears with normal hearing subjects using ILO 88 otodynamic analyzer system were measured, the click stimuli level is 80 dB SPL and the stimuli pattern used non-linear model. RESULT: Analysis of the normal values of the stability, A-B difference, response amplitude, wave reproducibility, band reproducibility and band SNR, dominant frequencies were obtained. CONCLUSION: The findings from comprehensive analysis of spectrum, response amplitude and reproducibility can be used as main sensitive parameters to evaluated whether or not of normal-hearing. PMID- 12541344 TI - [Measuring of spontaneous otoacoustic emissions of adolescence and its significance]. AB - OBJECTIVE: In order to study the relationship between spontaneous otoacoustic emissions (SOAE) and cochlear function. METHOD: SOAE of 50 normal hearing young people were measured. The incidences, amplitudes and frequencies of SOAE were obtained. RESULT: The SOAE incidence was 40% in 100 ears of 50 cases and there was no significant difference in sexuality and laterality (P > 0.05); In the 40 cases who had SOAE, the amplitudes ranged from--27.5 dB SPL to 9.7 dB SPL and the frequencies ranged from 269 Hz to 5,660 Hz, without sexual difference; There is no significant difference between SOAE and threshold of pure tone acoustic (P > 0.05). CONCLUSION: The appearance of SOAE indicated good cochlear function. PMID- 12541345 TI - [The normal value of DPOAE and effect of intensity differ of probe tone]. AB - OBJECTIVE: To investigate the normal value of DPOAE and effect of intensity differ of probe tone. METHOD: 30 subjects (60 ears) age from 17 to 40 were examined in this article, and the amplitude of DPOAE were measured when using L1 = L2 = 70 dB SPL and L1 = 70 dB SPL, L2 = 65 dB SPL. RESULT: DPOAE was recorded in 100%. The amplitude of DPOAE when using L1 = 70 dB SPL, L2 = 65 dB SPL is larger than the amplitude of DPOAE when using L1 = L2 = 70 dB SPL. CONCLUSION: Those results give us useful information for clinic. PMID- 12541346 TI - [The selection of tympanoplastic type in treating chronic suppurative otitis media]. AB - OBJECTIVE: To investigate the effect of different type of tympanoplasty on chronic suppurative otitis media (CSOM). METHOD: Myringoplasty, focuses clearance and myringoplasty, focuses clearance and ossiculoplasty and myringoplasty were performed in 189 cases of CSOM. RESULT: The cure rate of perforation of tympanic membrane with myringoplasty was above 99% accompanied by the varying improvement in acoustic sensibility. CONCLUSION: Different types of tympanoplasty should be used according to the CSOM. The slighter the pathological changes of CSOM, the more the hearing improve. The better effects was in tympanoplasty of simple type. The tympanic exploration is necessary to be included in tympanoplasty. PMID- 12541347 TI - [Effect of ear-canal air pressure variation on tempanometry using a sweep frequency probe tone]. AB - OBJECTIVE: By observing the effect of ear-canal pressure variation (ECPV) on tempanometry using a sweep-frequency probe tone, the roles of sweep-frequency tempanograms changes were studied. METHOD: Tympanograms using a sweep-frequence probe tone were measured for 60 ears of 30 subjects at ECPV by admittance (Y), susceptance (B), conductance (G) with GSI33 middle ear analyzer. RESULT: There was not obviously difference about phase angle and resonance frequency among 3 measured conditions (P > 0.05); Resonance frequency became higher with higher positive pressure and lower with lower negative pressure in ear-canal; delta Y, delta G got lower with higher positive and lower negative pressure at resonance frequency; delta B is about 0 mmhos and almost didn't change with ECPV; phase angle decreased with higher positive or lower negative pressure. delta Y, delta G and delta phi were the highest at the peak pressure. CONCLUSION: The higher the ear-canal positive pressure or the lower the negative pressure is, the higher the compliance and the lower the mass of middle ear is comparatively. The rub of middle ear is the lowest at normal peak pressure. PMID- 12541348 TI - [Modificated epiglottic laryngoplasty]. AB - OBJECTIVE: To investigate the surgical method and effect of laryngeal rehabilitation of patients with glottic carcinoma (T3 category) after extended partial laryngectomy. METHOD: 36 cases were operated with modificated epiglottic laryngoplasty. The materials for laryngoplasty included the epiglottic and one lateral partial thyroid cartilage and muscle-cartilage membrane flap and one arytenoid cartilage. The modificated technique was designed. RESULT: 33 cases were postoperatively in pulling-out-tube. The voice of 30 cases was clear, loud and comprehensible, but low and hoarse voice in other 3 cases. The 3 and 5 year survival rates were 89.3% and 77.3% respectively. The 3 and 5 year stable rates of laryngeal function were 95.6% and 94.1% respectively. CONCLUSION: The modificated epiglottic laryngoplasty was effective surgical method for the patients with glottic carcinoma (T3 category). PMID- 12541349 TI - [Clinical manifestations and CT diagnosis of non-allergic fungal sinusitis]. AB - OBJECTIVE: The diagnose of fungal sinusitis, especially complicated by bone destructive, and its differential diagnosis from malignant tumour of nasal sinus were studied. METHOD: Five cases were diagnosed with CT; nasal sinus microsurgery was done in 4 cases and sinusotomy in one case. None of them were treated by antifungal agents. RESULT: Fungal filaments were postoperatively shown in 5 pathological reports, fungal spores in some cases. No recurrence and complications occurred as followed from 6 months to 3 years. CONCLUSION: It is not easy to differentiate fungal sinusitis from malignant tumour of nasal sinus, sometimes, but CT has demonstrated its importance of diagnostic value. PMID- 12541350 TI - [Malignant melanoma of external ear: 14 cases report]. AB - OBJECTIVE: To improve the diagnosis and treatment level of malignant melanoma in external ear. METHOD: 14 cases with malignant melanoma of external ear were reported. All patients were treated by surgical excision. RESULT: 13(in 14) cases were followed up. 7 patients were survival with no tumor, the others died. The 5 year survival rate for the whole series was 50.0%. CONCLUSION: The malignant melanoma of external ear has rapid progress and poor prognosis, early detection and treatment are necessary. Extensive excision of malignant melanoma was principle method of the treatment. Recurrent tumor should be treated by combined operation with radiation. PMID- 12541351 TI - [The dermoid cyst of maxillofacial suture]. AB - OBJECTIVE: To investigate the clinical course, pathologic characteristic, diagnosis and surgical treatment of dermoid cyst of maxillofacial suture. METHOD: Eight cases of dermoid cyst of maxillofacial suture were reported. RESULT: All patients were treated by surgery. The cure rate was 100%. No recurrence was found as follow-up from 1 to 4 years. CONCLUSION: There are some features in the clinical course of the dermoid cyst of maxillofacial suture. It is necessary to do CT examination. Surgery operation is the only treatment by now. PMID- 12541352 TI - [Endoscopic treatment of sphenoidal and ethmoidal mucoceles]. AB - OBJECTIVE: To evaluate endoscopic surgery for sphenoidal and ethmoidal mucoceles. METHOD: Seventeen patients diagnosed as having paranasal sinus mucoceles underwent attempted endoscopic marsupialization between 1989 and 1995. Ten of the mucoceles were in the sphenoid sinus, seven in the ethmoid sinus. Postoperative endoscopic follow-up period varied from 3 to 9 years. RESULT: All the sphenoidal and ethmoidal mucoceles were satisfactorily approached endoscopically. Postoperative endoscopic examination manifested all the operating cavity was satisfactorily marsupialized. The symptoms were improved or relieved in 15 patients. None of the patients undergoing endoscopic surgery had significant complications. CONCLUSION: Our results show that endoscopic surgery seems to be a safe, valuable way for management of sphenoidal and ethmoidal mucoceles. PMID- 12541353 TI - [An electrophysiological study of the kainic acid toxicity on cochleas]. AB - OBJECTIVE: The purpose of this study was to investigate the kainic acid (KA) toxicity on cochleas. METHOD: One microliter KA in the concentration of 60 mM was applied to the round window membrane of guinea pig. The compound action potential (CAP), cochlear microphonic potential (CM) and distortion product otoacoustic emissions (DPOAE) were measured 12 hours after KA was administrated. RESULT: CAP was suppressed significantly or abolished after the drug administration, however, the CM and DPOAE almost presented no change. CONCLUSION: The targets of KA toxicity are the primary auditory afferent endings. PMID- 12541354 TI - [Cell infiltration in nasal polyps and it's pathological significance]. AB - OBJECTIVE: To study the relation between inflammatory cells infiltration in nasal polyp tissues and it's pathogenesis. METHOD: Nasal mucosa from 10 normal individual and 4 patients of nasal polyps were studied with immunohistochemical technique and histopathologic examination. RESULT: The number of eosinophils, CD68 positive cells and monocytes was significantly higher (P < 0.05) in nasal polyps tissues than in the normal controls. CONCLUSION: Infiltration of inflammatory effector cells such as eosinphils and chronic inflammatory changes of the nasal mucosa might play a role in nasal polyps sprouting. PMID- 12541355 TI - [Radiofrequency treatment of ethmoidal nerve with allergic rhinitis under nasal endoscopy]. AB - OBJECTIVE: To study the effect of radiofrequency treatment of ethmoidal nerve on allergic rhinitis. METHOD: Eighty-eight cases of allergic rhinitis were treated with ethmoidal nerve radiofrequency under nasal endoscopy. RESULT: It showed marked effectiveness in 73 cases (83.0%), immprovement in 11 cases (12.5%) and failed in 4 cases (4.5%) in one year follow-up. The overall effective rate was 95.5%. CONCLUSION: This result indicated that it is a choice for treating allergic rhinitis to apply ethmoidal nerve radiofrequency under nasal endoscopy. PMID- 12541356 TI - [Expression of vascular endothelial growth factor and tumor angiogenesis correlates with biological behaviors of nasopharyngeal carcinoma]. AB - OBJECTIVE: To study the correlation between tumor angiogenesis and biological behaviors of nasopharyngeal carcinoma(NPC). METHOD: Angiogenesis was assessed by the microvessel dinsity(MVD) and expression of vascular endothelial growth factor(VEGF) was detected using immunohistochemical S-P method. Paraffin-embedded specimens from 65 patients with NPC and 15 with benign nasopharyngeal tumors, 15 with tumor-free nasopharyngeal mucosac were detected. RESULT: Expressions of VEGF and MVD in NPC group were higher than that of nonmetastic carcinoma group. The relationship between expressions of VEGF and MVD and the clinical staging present statistic significance. CONCLUSION: MVD and expression of VEGF are highly correlated with biological behaviors of NPC, which may serve as a parameter for determining tumor biological metastatic potential and prognosis. Angiogenesis inhibitors might be helpful in preventing NPC from metastasis. PMID- 12541357 TI - [Surgical treatment of the functional obstruction of eustachian tube of middle aged and aging people]. AB - OBJECTIVE: To expore the effect and the way of palatosapingeus tendon-shortening surgery to treat the functional obstruction of eustachian tube of middle-aged and aging people. METHOD: Seven cases of palatosapingeus tendon-shortening surgery were analyzed retrospectively. RESULT: After operations the symptoms of all cases were relieved in a week. The tympanogram returned to type "A". The audiogram showed that the airbone gap disappeared. The shape of tympanic membrance returned to normal state. None of them recured in one-three year following up. CONCLUSION: The effect of palatosapingeus tendon-shortening surgery is satisfactory so long as its surgical indication is correctly selected. PMID- 12541358 TI - [Relationship between expression of estrogen receptor progestrone receptor and the biological characteristics of nasopharyngeal carcinoma]. AB - OBJECTIVE: To Clarity the relationship between the expression of the estrogen receptor (ER), progestogen receptor (PR) and the biological characteristics of nasopharyngeal carcinoma (NPC). METHOD: The immuno-histochemical LSAB method was used to analyse the ER and PR in paraffin section specimen of 71 cases of nasopharyngeal carcinoma. CT scaning and clinic presentation were also put into consideration. RESULT: The expression of ER with (-), (+), (++) and (+++) in 71 cases were: 4(5.63%), 19(26.76%), 27(38.03%) and 21 (26.76%), respectively. The expression of PR with (-), (+) and (++) in 71 cases were: 41(57.75%), 29 (39.44%) and 2(2.82%), respectively. The encroaching of tumor to the tissue of parapharynx, the metastasis rate of lymphonode and clinic stage were related to the expression of ER (P < 0.01). The encroaching of tumor to the tissue of parapharynx, the metastasis rate of lymphonode were also assciated with the expression of PR (P < 0.05). CONCLUSION: NPC is a tumor of hightly expression of ER and PR. The positive intensity of ER and PR were associated with the encroaching extent of NPC, and the metastasis rate of lymphonode. It is significant to detarmine ER and PR of NPC in assessing the biological characterisitics of NPC and predicting the prognosis of NPC. It also suggests that the endocrine therapy can be put into consideration. PMID- 12541359 TI - [Applying combination of ultrasound and CT to diagnose paralaryngeal space tumor]. AB - OBJECTIVE: To explore an effective way to diagnose parapharyngeal space tumor. METHODS: The clinical data of 25 cases of this type of tumor, diagnosed by combinating ultrasound examination and CT scan, were analysed. RESULTS: The diagnosis correspondence rate by the combinating way of ultrasound and CT is 92%. The combination of the two methods is likely able to reveal the overall shape, extend and feature of tumor, and to assist us in decision of the entrance of operation. CONCLUSION: The combination of ultrasound diagaosis and CT scan is of satisfactory clinical value in diagnosis of parapharyngeal space tumor. PMID- 12541360 TI - [Diagnostic value of ultrasonography in cervical lymph node metastasis]. AB - OBJECTIVE: To research the accuracy of ultrasonography in the diagnosis of cervical lymph node metastasis and the value of that of cervical lymph node occult metastasis. METHOD: Fifty-five patients(61 sides) operated by neck dissection were double-blindedly studied by preoperation palpation and ultrasonography comparing with postoperating pathological examination. RESULT: Fifty-two sides were confirmed to be pathology lymph node metastasis from 61 sides. 40 and 48 sides were checked out by palpation and ultrasonography, respectively. The sensitivity, specificity and accuracy of palpation and ultrasonography were 76.9% and 92.3% (P < 0.05), 77.8% and 88.9%(P > 0.05) and 77.0% and 91.8%(P < 0.05), respectively. Six sides were checked out by ultrasonography (occult metastasis) in 12 sides of palpation negative. CONCLUSION: The accuracy of ultrasonography was prior to that of palpation in examining cervical lymph node metastsis. Ultrasonography could check out about half occult metastasis and should be regarded as a routine examining items in diagnosing cervical lymph node metastasis. PMID- 12541361 TI - [Experience of endoscopic sinus operation on 74 cases]. AB - OBJECTIVE: To improve the curative effect of endoscopic sinus operation. METHOD: The clinical data of nasal cavity and nasal sinus via endoscopic sinus operation on 74 cases (115 sides) were in summary. Of those 74 cases, 65 were followed-up over six mothes. RESULT: It showed that 44 cases were cured, 18 were improved, 3 were ineffective. The effective rate was 95.4%. The disappearance rate of four symptons-nasal obstruction, headache, purulent nasal discharge and loss of smelling was 95.2%, 92%, 66.7% and 42.5%, respectively. The serious operative complication was absent. CONCLUSION: It helped to raise thd effect of anesthetization to apply nasal cavity surface anesthesia, nerve block anesthesia. Paying attention to the distinguish of dissection label can help to decrease the occurance of the serious complication of the sinus ethmoidal operation. The operation which was alternatively conducted between endoscope and cavascope, according to the operation requirement, can be carried out more thoroughly and more safely. Cleaning the operation cavity after operation can promote healing. PMID- 12541362 TI - [Study of the relation between MMP2, MMP9 and nasopharyngeal carcinoma]. AB - OBJECTIVE: To evaluate the role of matrix metalloproteinase (MMP) in the invasion and metastasis of nasopharyngeal carcinoma (NPC). METHOD: Twenty-six cases of NPC were studied by immunohistochmical method using monocolonal antibody to MMP2 and MMP9. RESULT: It was showed that there were significantly correlation between MMP2 or MMP9 and the invasion and metastasis of NPC. The positive rate of MMP2 and MMP9 were higher in NPC with cervical lymph node involvement than that of no cervical lymph node involvement. And they were lower in well differentiated NPC than poorly differentiated NPC. CONCLUSION: The results indicate that MMP2 and MMP9 may play a critical role in assisting carcinoma cell to degrade basement membrane to invade surrounding tissues and to form metastatic colonies in lymph nodes. PMID- 12541363 TI - [Effects of acoustic stimuli on intracellular free calcium isolated cochlear OHC]. AB - OBJECTIVE: To study the effects of acoustic stimuli upon intracellular free calcium concentration ([Ca2+]i) of cochlear outer hair cells (OHCs) isolated from guinea pigs. METHOD: The specimens were stained with fluorescence, and measured by using ACAS 570. RESULT: Acoustic stimuli caused a remarkable increase of [Ca2+]i and the concentration of [Ca2+]i decreased gradually after reaching its peak value. The reponeses can repeat with another stimuli. CONCLUSION: The acoustic stimuli could alter the physiologic characteristics of single OHC. PMID- 12541364 TI - [The influence of nasal blood flow after ligating external carotid artery and its branches in rabbits]. AB - OBJECTIVE: To study the influence of nasal blood flow after ligation of external carotid artery and its branches. METHOD: We ligate different levels of external carotid artery and its branches, and use laser Doppler flowmetry to observe the change of blood flow in nose before and after operation. RESULT: Among the four types of operation ligating different blood vessels to reduce the blood flow of the operating side, the most effective way is to ligate ipsilateral carotid artery and its branches in the same side. CONCLUSION: After ligation of relating blood vessels, the collateral circulation construct very soon, and the founding speed of that is quicker in the early stage. PMID- 12541365 TI - [Changes of CGRP activity and distribution in facial nucleus after facial nerve injury of guinea pig]. AB - OBJECTIVE: In order to study the changes of CGRP activity in facial nucleus of guinea pig after facial nerve injury. METHOD: The activity of CGRP was detected by the technique of immunohistochemical and image analysis methods were used. RESULT: It was showed the there were CGRP-immunoreactivity (IR) in normal facial nucleus and its' subnucleus. An initial increase of CGRP-IR was noted and the content of CGRP under image analysis were significantly different when compared with control group (P < 0.01) at day 1 after injury of facial nerve. CGRP continued to rise to a maximal level at day 7 (P < 0.001). And after which it gradually decreased. We noticed distinct bundle-shaped CGRP fibers elongated to the peripheral part from ventral facial nucleus in the period of 14 d-35 d. CONCLUSION: It suggests that during the course of facial nerve regeneration following injury, CGRP changed characteristically. CGRP may play the regulational role in facial nerve regeneration. PMID- 12541366 TI - [With periosteal "sandwich" suturing and repairing septonasal perforation by nasal columella degloving]. AB - OBJECTIVE: To recommend a new type of repairing septonasal peforation. METHOD: After a seagull's incision is made around the internal edges of anterior nares and the root of nasal columella, the mucoperiost of nasal septum and nasal bottom is widely dissected through the nasal columella degloving pathway. Then, relaxative incisions are made in bilateral mucoperiost at different directions and bilateral peforations are relaxative sutured at different levels respectively. Meanwhile, an autograft of tibil periost is fixed into nasal septum by the "sandwich" technique. Finally, both nasal cavities are carefully packed. RESULTS: Three cases of septonasal perforation were successfully repaired and have been followed up for 1-2 years respectivelly with no recurrence. CONCLUSION: The characteristics of this operation are: style rather fashionable, pathway rather wide, mucoperiost rather relaxed, suture rather firm and failing rather few. PMID- 12541367 TI - [Studies on human nasal cycle with acoustic rhinometry]. AB - OBJECTIVE: The aim of this article was to study the classifications and characteristics of nasal cycle of adult and children and the influence of pathological state of nasal mucosa on nasal cycle with acoustic rhinometry. METHOD: We examined the nasal cycle of 16 healthy adults, 11 healthy children and 14 patients with bilateral nasal obstruction. Acoustic rhinometry was performed every 30 minutes (20 minutes in children) over 4-8.5 hours to determine minimum cross-sectional area of nasal cavity (MCA) and nasal cavity volume (NV) which were used as indices of nasal patency. RESULT: Nasal cycles were observed in all subjects. Classical nasal cycle was most commonly observed in healthy adults and patients with nasal diseases. In healthy children, in concert nasal cycle was the most common type. The distribution of the healthy adults and patients with nasal diseases between the cycle categories was not statistically significant different (P > 0.05), the mean periodicity of the cycle was no significantly different between these two groups (P > 0.05). The mean alteration amplitude of nasal cycle of healthy adults was significantly longer than that of patients with nasal diseases (P < 0.01). The distribution of the healthy children and the healthy adults between the cycle categories was significantly different (P < 0.01), the mean periodicity of the cycle of healthy children was distinctively shorter than that of healthy adults (P < 0.01). CONCLUSION: The above results illustrated that nasal cycle exists as central control and local modulation. In healthy children, in concert nasal cycle was most commonly observed and this may be related with the immature condition of their autonomic nervous system. PMID- 12541368 TI - [Anatomic investigation of facial canal related to middle ear surgery]. AB - OBJECTIVE: To avoid hurting facial nerve in middle ear surgery. METHOD: 20 adult head cadavers with 40 sides tempotal bone were dissected and investigated. RESULT: 1. Prevalence of dehiscent facial nerve canal was 32.5% (13/40), while 2.5% (1/40), 27.5% (11/40), 7.5%(3/40) incidence facial nerve canal were found in the labyrinthine, horizontal, vertical segment respectively. 2. Distance between the vertical portion of facial nerve and posterior semicircular canal was 0.62 5.08(2.48 +/- 0.63) mm. Interspace between the vertical portion of facial nerve and tympanic chorda was 0-4.62 (2.12 +/- 0.76) mm at the level of tympanic ditch, while interspace between the vertical portion of facial nerve and tympanic ditch was 2.96-8.82 (5.86 +/- 1.62) mm. 3. Distance between horizontal portion of facial nerve and lateral semicircular canal was 0-3.02 (1.92 +/- 0.84) mm. Interspace between horizontal portion of facial nerve and stapedial board was 0.92-4.06(2.14 +/- 0.90) mm. CONCLUSION: Familiarity with facial nerve procession should be favorable to decrease facial paralysis with middle ear surgery. PMID- 12541369 TI - [Influence of pavulon on facial nerve monitoring during middle ear-mastoid operation]. AB - OBJECTIVE: To evaluate the influence of pavulon (muscle relaxant) on facial nerve monitoring for patient with middle ear-mastoid operation during general anesthesia. METHOD: Intraoperative monitoring of the facial nerve (FNM) was done in 45 ears (44 case). For patient with middle ear-mastoid disease, 38 ears underwent inhaled-intravenous anesthesia with muscle relaxant pavulon (group of general anesthesia), 7 ears underwent local anesthesia (group of local anesthesia). Accelography (ACCG) was used in 10 case for testing the correlation between of FNM and the various degree of recovery from ulnar nerve-musculor blockade. RESULT: FNM could be elicited during different periods of recovery with values of T4/T1 varying from 10%-98%. Pavulon could be used to eliminate muscular vibration from disturbing FNM. Electromyography (EMG) potentials were successfully recorded in 34 ears (89.4%). Anesthesia was uneventful and it may be controled satisfactority. CONCLUSION: Considering the advantage of pavulon. It can be used for general anesthesia during intraoperative monitoring of the facial nerve for children or adults suffering from local complicated diseases. PMID- 12541370 TI - [Transantral orbit decompression to malignant ophthalmopathy]. AB - OBJECTIVE: To recognize the indication and the value of transantral orbit decompression to malignant ophthalmopathy. METHOD: 9 eyes of 5 cases undergone transantral orbital decompression from November 1986 to October 1998 were reviewed. RESULT: After 3 months-2 years of follow-up. We found this approach successfully improved visual function of all 5 cases. Average proptosis reduction was 5 mm. The eye lids could be closed in 4 of the 5 cases and the palpebral fissures was decreased 1-2 mm in 1 of the 5 cases. Preoperative exposure keratitis resolved in both of the 2 cases. Diplopia was cured in both of the 2 cases. Secondary glucorna also recovered in both the 2 cases. Disturbance of ocular motility in both of the 2 cases improved postoperatively and preoperative visual acurity worsen also improved in both of the 2 cases. CONCLUSION: It has been suggested that the transantral decompression of the orbit provided a safe, effective therapeutic modality for patients with vision-threatening or major cosmetic problem. PMID- 12541371 TI - [Analysis of 16 cases with aggressive fungal maxillary sinusitis]. AB - OBJECTIVE: To improve the diagnosis and treatment level of aggressive fungal maxillary sinusitis. METHOD: 16 cases were diagnosed with CT. Caldwell-Luc's surgery was done in 13 cases and Lima surgery in 3 cases. None of them were treated by antifungal agents. RESULT: It showed osteolytic damage with bony defect which was different from expanded bony defect caused by benign tumor, but similar to malignant tumor. No recurrence occurred as followed from 6 months to 2 years. CONCLUSION: Computed tomography is very important in its diagnosis. Early explorative operation is useful in making definite diagnosis. PMID- 12541372 TI - [Applied anatomy of anterior ethmoid relevant to endoscopic transnasal dacryocystorhinostomy]. AB - OBJECTIVE: This study provided clinical guidance for the endoscopic transnasal dacryocystorhinostomy. METHOD: 16 adult cadavers were studied on the applied anatomy of between anterior ethmoid and fossa of lacrimal sac and the anatomic relationship between uncinate process and medial wall of orbit. RESULT: According to the anatomic relationship between anterior ethmoid sinus and fossa of lacrimal sac, the cells of anterior ethmoid sinus were classified into three typies, The results was that type I accounted for 31.2%, type II 50.0% type III 18.8%. CONCLUSION: CT scans should be performed to determine the anatomic relationships between fossa of lacrimal sac and anterior ethmoid and between uncinate process and medial wall of orbit before operation for patients with different typies of obstruction of nasocrimal duct, thus the proper bony opening was located during dacryocystorhinostomy. PMID- 12541373 TI - [Clinical study of capsaicin in the treatment of allergic rhinitis]. AB - OBJECTIVE: To study the clinical effect of capsaicin of nasal administration in 100 cases of allergic rhinitis. METHOD: Capsaicin was applied by nostril, once 5 days and 5 times as a therapeutic course. Acidophil were measured and nasal airflow resistance was analysed before and after capsaicin therapy. RESULT: It showed that nasal symptoms of patients were obviously relieved and 94% of the cases in the measure of acidophil changed to negative from positive. Moreover, capsaicin induced decrease in nasal airflow resistance from (0.6272 +/- 0.3534) kPa.s/L before therapy to (0.4057 +/- 0.2419) kPa.s/L after therapy (P < 0.001). 90 cases of 100 patients were fellowed up one year, the total effective rate was 89%. CONCLUSION: Our study suggests that capsaicin possessed remarkable effects in the treatment of allergic rhinitis; analyse of nasal airflow resistance may be used as one of the objective guideline in the determinant of therapeutic effects. PMID- 12541374 TI - [Clinical study of treatment of epistaxis of little's area by microwave, anterior nasal packing and trichloroacetic acid cautery]. AB - OBJECTIVE: To study the difference and quality of three different kind of treatment methods for epistaxis of Little's area, such as: microwave, anterior packing of nasal cavity and trichloroacetic acid cautery. METHOD: The cases were divided into three group. The state of bleeding, reaction of after-treatment and recurrence were recorded and analysed. RESULT: Comparing with the anterior nasal packing and trichloroacetic acid cautery, the microwave is characteristic by thorough stop-bleeding, light reaction, less recurrence. CONCLUSION: Microwave is an ideal way to treat epistaxis of Little's area. The patient's reaction is mild and less the clinical effects is obviously better than the anterior packing of nasal cavity and trichloroacetic acid cautery. PMID- 12541375 TI - [Clinical analysis of 28 patients with unsuccessful septoplasty]. AB - OBJECTIVE: To analyse the causes of unsuccessful septoplasty. METHOD: 28 cases were investigated by high-resolution CT scan and nasal endoscopy. Another operation was completed in endoscopic sinus surgery. RESULT: 16 upper or posterior nasal septal deviation and 12 paraseptal structural deformities and chronic sinus diseases were observed, which were neglected in the first operation. CONCLUSION: Before septoplasty, CT and endoscopic evaluation of the paranasal sinuses and paraseptal structures should be performed and accompanied nasal diseases should be considered. PMID- 12541376 TI - [Effection of children airway foreign bodies on blood acid-base disturbances]. AB - OBJECTIVE: To explore the effects of children airway foreign bodies on blood acid base disturbances. METHOD: To detect the blood-gas and electrolyte of 50 cases by children airway foreign bodies (test group), they were compared with 50 cases normal children (control group). To calculate anion gap (AG), to determine types of acidbase disturbances. RESULT: The blood-gas and K+ were significant difference between that two groups. In the test group, there were 25 simple and 19 mixed acid-base disturbances; 6 acid-base balances. There were 31 high AG, 18 normal AG, one low AG. CONCLUSION: Children airway foreign bodies can cause acid base disturbances. It was major in simple metabolic acidosis and mixed metabolic acidosis with respiratory alkalosis, it was more in high AG-metabobic acidosis. PMID- 12541377 TI - [Characterization of nitric oxide synthase in the chicken cochlea using electrochemical method]. AB - OBJECTIVE: Our investigation was aimed to explore whether certain agonists can induce NO release from neurons of chick cochlea in vitro to characterize NOS isoform in the cochlea. METHOD: The experiments were performed using CuPtCl6 chemically modified carbon fiber microelectrode under current recording in terms of NO redox activity. RESULT: In our previous study it has been shown that solutions containing L-arg, ATP, ACh with Ca2+ can mediate NO release from neurons of chick cochlea, respectively. The present study examined visual current changes induced by other stimuli to cochlear neurons. The solution containing glutamate with Ca2+ could exerted a positive effects and L-arg, ATP, ACh, glutamate without Ca2+ and bradykimin with Ca2+ or without Ca2+ can not induce NO release from the neurons, respectively. CONCLUSION: Based on these results, the NOS isofrom in the chick cochlear neurons is the neuronal isofrom (ncNOS). PMID- 12541379 TI - [Epidemiological survey on orchard pollinosis]. AB - OBJECTIVE: To investigate the prevalence of Orchard Pollinosis in China. METHOD: From 1995 to 1998, an epidemiological survey on orchard pollinosis was carried on among 1,660 primary and middle school students and 2,167 university students, including questionnaire investigation, nasal inspection and scratch test. From April to June of 1998, a clinical investigation of orchard pollinosis was adopted among 30 patients of allergic rhinitis including scratch test and nasal mucosa test. RESULT: 1. The total positive rate of orchard pollen scratch test was 6.7%, and the prevalence of orchard pollinosis was 0.37%. 2. 5 cases (16.7%) were hypersensitive to orchard pollen clinically in allergen test. CONCLUSION: Orchard pollinosis is one kind of important pollinosis and should be paid high attention to. PMID- 12541378 TI - [Cochleogram for assessing hair cells and efferent fibers in carboplatin-treated ear]. AB - OBJECTIVE: This paper describes a convenient histochemical technique that allows one to simultaneously assess the cochlear efferent neurons and inner (IHCs) and outer hair cells (OHCs). METHOD: Selective labeling of the sensory cells and efferent fibers was carried out by first labeling the hair cells for dehydrogenase activity and then staining the efferent fibers for acetylcholinesterase activity. Double-labeled specimens were prepared as a surface preparation and counts were made of the number of hair cells and the number of discreate fiber bundles crossing the tunnel of Corti along the entire length of the cochlea. RESULT: The utility of this technique for assessing ototoxic damage was demonstrated by constructing cochleograms showing the percentage of missing hair cells and tunnel-crossing efferent fibers in chinchillas treated with carboplatin. In the region of the cochlea associated with most IHC loss and a part of OHC missing showed by succinate dehydrogenase in the region of hair cells; however, the loss of tunnel-crossing efferent fibers showed by acetylcholinesterase staining was closely correlated with the percentage of OHC loss. But, if only damage the IHC, the efferent fibers showed normal. CONCLUSION: These results suggest that carboplatin damaged efferent systems may be due to the damage of OHC. PMID- 12541380 TI - [The detection and clinical significance of telomerase activity in supraglottic carcinomas]. AB - OBJECTIVE: To study the expression and significance of telomerase activity in laryngeal carcinoma. METHOD: We detected telomerase activity of HEP-2 cell line, 26 cases of supraglottic carcinoma tissues and 15 cases of peri-carcinoma tissues with PCR-based assay designated TRAP (for telomeric repeat amplification protocol). RESULT: The telomerase positive rate of supraglottic carcinoma 84.6% (22/26) was obvious higher than that of peri-carcinoma 40% (6/15), P < 0.01. No significant differences in positive rates were found among different groups divided according to clinical stage or metastasis of cervical lymph node, P > 0.05. CONCLUSION: Telomerase activation is related to immortalization process, though not much helpful in diagnosis of supraglottic carcinoma. PMID- 12541381 TI - [Evaluation of N staging of cervical lymph nodes with positive palpation in laryngeal carcinoma]. AB - OBJECTIVE: To evaluate N staging of cervical lymph nodes with positive palpation objectively. METHOD: Serial sections of lymph nodes of neck dissections with transparent methods were observed under light microscope. RESULT: 81 lymph nodes with pathological metastases were verified in 100 patients with 114 palpation positive lymph nodes. The correspondent rate was 71.1%. The correspondent rate of clinical N via pathological N staging was N1 35.6%, N2a 25.0%, N2b 58.3%, N2c 75.0%, N3 100.0% respectively. CONCLUSION: N staging judged only by palpation is not sufficient in clinic. It should be combined with other examinations. PMID- 12541382 TI - [The value of nasal endoscopy in the resection of upper and posterior nasal septum deviation]. AB - OBJECTIVE: In order to find a safe, reliable and satisfying technique to treat the upper and posterior nasal septum deviation. METHOD: Performing the resection of the upper and posterior nasal septum deviation under nasal endoscopy, comparing this new method with the traditional one and discussing the safety and long-term curative effect of this new method. RESULT: The observation of the outcome of 109 patients undergoing this operation after 1-50 months postoperation shows that this technique can resect the upper and posterior deviation safely and completely, improve the nasal respiratory and olfactory function and reduce the occurrence of nasal cavity adhesion and the recurrence of nasal polyp and sinusitis. CONCLUSION: Performing the resection of the upper and posterior nasal septum deviation under nasal endoscopy is a good method for popularizing to clinician. PMID- 12541383 TI - [Immunohistochemical study of safe resection margin in the surgical treatment of laryngeal carcinoma]. AB - OBJECTIVE: To study safe resection margin of laryngectomy according to the level of protein of gene expression. METHOD: Using citric-LSAB-immunochemical technique, the expressions of ras, c-myc and P53 were studied in different regions of larynx including 30 cases laryngeal carcinoma, border area, adjacent mucosa which was 0.5, 1.0, 1.5, 2.0 cm away from cancer and four normal laryngeal mucosa. RESULT: The results showed that both single expression and co-expression of p21, p62 and p53 increased as tissue progressed in sequence from normal mucosa, 2.0, 1.5, 1.0, 0.5 cm distant mucosa adjacent to carcinoma, border area and carcinoma. A significant different expression in most of them can be seen at regions of 0.5 cm and 1.0 cm distant from the tumor (P < 0.05). CONCLUSION: The tissue 0.5 cm adjacent to the tumor should be regarded as precancerous lesion. It is appropriate to regard 0.5 cm away from tumors as resection margins for surgical treatment of laryngeal carcinoma. PMID- 12541384 TI - [Complications of PPP: prevention and management strategies]. AB - OBJECTIVE: In order to minimize the complications of palatopharyngoplasty (PPP). METHOD: Perioperative complications of PPP of 110 cases were analysed. RESULT: In our series, the complications included velopharyngeal insufficiency (28 cases), hypertensive crisis (12 cases), respiratory embarrassment (12 cases), wound dehiscence (2 cases), local hemorrhage (1 cases), hypovolemic shock (1 cases) and nasopharyngeal stricture (1 cases). CONCLUSION: It is concluded that prevention is important. Prevention strategies included: 1. Preoperative careful examination; 2. Using local anesthesia; 3. Intraoperative monitor ECG, blood pressure and oxygen saturation; 4. Intraoperative inbreathing oxygen; 5. Staunch blood thoroughly; 6. Postoperative proper treatment with steroid; 7. CPAP treatment should be applied before PPP in the severe OSAS. PMID- 12541385 TI - [Applied anatomic study on intraorbital and intraethmoidal parts of ethmoidal artery]. AB - OBJECTIVE: In order to provide anatomic data for operation of ethmoidal sinus and optic canal decompression. METHOD: Using microanatomic method, the intraorbital and intraethmoidal parts of ethmoidal arteries were observed and measured. RESULT: The length of intraorbital segment of anterior ethmoidal artery was 5.16 +/- 1.24 mm, and its diameter was 0.56 +/- 0.17 mm; the length of intraorbital segment of posterior ethmoidal artery was 9.08 +/- 2.29 mm, with diameter 0.37 +/ 0.14 mm. The ethmoidal artery, nerve and vein were surrounded by fascial sheath which could extend to ethmoidal sinus. Three variants of ethmoidal arteries could be found: It passed through bony canal; It went between the superior wall of ethmoidal cell and ethmoidal mucosa; One part went through bony canal, but the other did not. CONCLUSION: It was important to find ethmoidal artery in operation of ethmoidal sinus and optic canal decompression. PMID- 12541386 TI - [Applied anatomy on musclebone flap of hyoideumomohyoid to renovate vocal fold]. AB - OBJECTIVE: In order to provide anatomical basis for renovating vocal fold with muscle-bone flap of hyoideum-omohyoid. METHOD: It was measured that the anatomical structure correlative with superior belly of hyoideum-omohyoid, artery, vein and nerve of the muscle and vocal fold morphosis by 26(52 sides) cephalocevical sample dissection. RESULT: It was observed that the muscle is 74.8 mm in length, 8.4 mm in width of starting part, 9.6 mm in width of middle part, 4.4 mm in width of tendon part. Its blood supply are mainly from cricothyriod artery (51.9%) and superior laryngeal artery (25.0%) with an arterial trunk of 9.8 mm in length and 0.7 mm in diameter. The venous trunk is 17.4 mm in length and 1.1 mm in diameter. The muscle is innervated by the sub-broaches arising from hypoglossal nerve ansa, the trunk is 19.6 mm in length and 0.6 mm in diameter. CONCLUSION: It is possible that musclebone flap of hyoideumomohyoid to renovate vocal fold according to the study of the muscle morphology characteristic. PMID- 12541387 TI - [Analysis of results of fundamental frequency and voice parameter in healthy young people]. AB - OBJECTIVE: To study and analyzing the parameter values of voice in normal healthy young people. METHOD: In present paper, voice quality of 153 cases were collected and analyzed with Dr. Speech software. RESULT: Jitter was (0.18 +/- 0.07)%, Shimmer (1.60 +/- 0.74)%, HNR (Harmonics-to-Noise Ratio) (25.34 +/- 3.12) dB, SNR (Signal-to-Noise Ratio) (25.39 +/- 3.09) dB and NNE (Normalized Noise Energy( 16.95 +/- 3.57) dB. Average of fundamental frequency in male was (160.81 +/- 24.27) Hz, in female was (297.42 +/- 35.89) Hz and in all was (206.35 +/- 70.77) Hz. At the same time, the results of some correlative parameter about voice was taken. CONCLUSION: We considered that there were significant differences between gender about fundamental frequency in parameter values of voice. Three formant produced at 4.5 times, 8.64 times, 17.04 times of the fundamental frequency in male, at 2.07 times, 4.96 times, 7.28 times in female. This data was usefulness to evaluate voice results of normal or abnormal. PMID- 12541388 TI - [The expression of suppressor gene p16, p21 and p53 in nasopharyngeal carcinoma]. AB - OBJECTIVE: To evaluate the roles of p16, p21 and p53 protein expression in nasopharyngeal carcinoma (NPC). METHOD: The SP immunohistochemical method was used to detect the expression of p16, p21 and p53 protein in 116 cases of NPC and 15 cases of non-tumour nasopharyngeal tissue. RESULT: 1. The positive expression rates of p16, p21 and p53 protein were 64.7%, 45.7% and 90.5% in NPC respectively. There were significant differences between NPC and non-tumour group (P < 0.05). 2. The expressive rate of p16 protein was 70.6% in non-keratinizing type, 44.8% in undifferentiated type (P < 0.05). 3. The positive expression of p21 was correlated with the expression of p16 and p53. CONCLUSION: Low expressions of p16 and p21 protein may play an important role in carcinogenesis of NPC, and that of p16 has some relation to cell differentiation of NPC. p16 protein may co-operate with p21 protein in development of NPC. p21 protein is probably not produced by stimulating wild-type p53 protein. PMID- 12541389 TI - [Neurilemmoma of the nasal cavity and paranasal sinuses (report of 12 cases)]. AB - OBJECTIVE: To further understand the neurilemmoma of nose. METHOD: 12 cases of neurilemmomas of the nasal cavity and paranasal sinuses were treated successfully by operation, 2 cases of malignant change lesions were treated by extensive resection and radiation. RESULT: 11 in 12 cases were followed up. 9 cases were cured and no any serious complications, 2 cases of malignant tumor were died. CONCLUSION: Early stage diagnosis is important. CT scan should be made as essential examination, complete surgical resection is the choice of treatment for primary lesions; malignant change lesions should be treated by combined operation with radiation. PMID- 12541390 TI - [Ectopic meningiomas of the head and neck (report of 10 cases)]. AB - OBJECTIVE: To study the diagnosis and treatment of ectopic meningiomas of the head and neck. METHOD: A retrospective review was made of 10 patients with ectopic meningiomas of the head and neck, the pathological diagnosis was as follows: 2 case epithelial type, 2 case psammomatous type, 1 case transitional type, 1 case malignant meningioma and types of another 4 cases were unknown. RESULT: All patients were treated surgically, 4 patients with local recurrence were operated secondarily and no signs were observed with the follow-up of many years. CONCLUSION: Surgery was the choice of ectopic meningiomas of the head and neck. PMID- 12541391 TI - [A comparative study of different neuromuscular pedicle embedded in posterior cricoarytenoid muscle]. AB - OBJECTIVE: To compare the results and effect of different neuromuscular pedicle (NMP) embedded in posterior cricoarytenoid muscle. METHOD: In 22 dogs, combined vocal cord paralysis models were made. The first group (10 dogs) were embedded by sternohyoid muscle pedicle of ansa cervicalis, the second (10 dogs), accessory sternocleidomastoid neuromuscular pedicle, the third (2 dogs), control group. RESULT: Evaluation was possible in 19 dogs. Reinnervation of the left posterior cricoarytenoid muscle occurred in 17 dogs following NMP. Return of spontaneous abduction in the operated side with NMP was noted after 4 months postoperatively. There was little difference between the two NMP groups. CONCLUSION: NMP was successful to reinnervate the posterior cricoarytenoid muscle. PMID- 12541392 TI - [Diagnostic significance of transiently evoked otoacoustic emissions in meniere's disease]. AB - OBJECTIVE: To judge the diagnostic significance of TEOAE by glycerol test in Meniere's disease. METHOD: TEOAE were recorded from all testing ears with or without Meniere's disease. RESULT: The wave reproducibility, amplitude of response, band reproducibility, band SNR in ears of affected ears with meniere's disease after administration of glycerol were increased, about which the parameters of affected ears in positive glycerol test were obviously higher than that in negative glycerol test. All parameters in normal ears with Meniere's disease increased too, and there have the changes of main frequency. CONCLUSION: TEOAE records of glycol test may be more sensitive and more simple than pure tone audiometry in diagnosing meniere's disease, and should be used more extensively. PMID- 12541394 TI - [A clinical study on sudden deafness in juveniles]. AB - OBJECTIVE: To understand the clinical characteristics, treatment efficacy and the main factors affecting the prognosis of sudden deafness in juveniles. METHOD: Clinical data of thirty-four cases about juvenile with sudden deafness were respectively analysesed. RESULT: 1) The number of male cases is as about four times as that of female cases, nearly one-third of the cases had a history of virus infection. 2) The total effective rate was 52.4%. The significantly differences among the different modalities were not found. 3) An association was found between the prognosis and the sex, the type of primary audiogram, the course of disease and the degree of hearing loss. The threshold of hearing at 4,000 Hz. was found valuable to judge the prognosis of sudden deafness in juveniles. The influence of the vertigo and tinnitus was not remarkable. CONCLUSION: Some aspects of sudden deafness in juvenile were similar to that in adult, however, there are also some special characteristics about it. PMID- 12541393 TI - [The observation of mycology and clinical efficacy in 325 cases with otomycosis]. AB - OBJECTIVE: To study otomycosis of pathogenic fungi, antifungal susceptibility test and clinical efficacy. METHOD: The study was performed as follows: 1) culture of pathogenic fungi and fungus identification in 110 cases. 2) antifungal susceptibility test with Etest strip by ketoconazole (KE), itraconazole (IT), Fluconazole (FC), Fluorocytosine (FL), and amphotericinum B (AP). 3) Topical application of doktarin ps cream (DC), ketoconazole cream (KEC), clotrimazole ung (CU), and thymol alcohol (TA) were performed in 4 groups and the intermittent impulsive treatment in who failed to respond to local treatment were performed too. RESULT AND CONCLUSION: 1) Chiefly, otomycosis of pathogenic fungi were aspergillus (79%) and candida albicans (8%) and penicillium citrinum (5%). 2) Antifungal susceptibility test showed that aspergillus were sensitive to KE and IT, but not sensitive to KE and FL, and candida albicans were sensitive to KE, IT, FC, FL and AP all; P. citrinum was sensitive to AP, FL and KE but not to FC. 3) Effective rate of topical application on 4 groups were 97.6%, 97.5%, 90.0% and 80.0% respectively. Comparing the results between compound azoles drugs and TA, there was a statistical significant difference (P < 0.05), the efficacy of compound azoles is better than that of TA. The effective rate of systematic intermittent impulsive treatment in 30 cases failed to respond to local treatment were 100%. PMID- 12541395 TI - [Observation of distortion product otoacoustic emissions in recovery course of sudden deafness]. AB - OBJECTIVE: To realize the DPOAE resulting in sudden deafness and to investigate the changes of OAE in the recovery course of sudden deafness. METHOD: The basic characters of distortion product OAE (DPOAE) were studied in 60 ears of 30 cases with sudden deafness. The treatment and following-up were carried on too. RESULT: The incidence and amplitude of OAEs in the affected ears were lower than that of normal ears and normal group, and the threshold of DPOAE was higher too. In the recovery course the amplitude and threshold of DPOAE were improved with the restoration of auditory threshold, but the restoring rate (RR) of auditory threshold was higher. CONCLUSION: The DPOAE in sudden deafness implies a serious sensorineural hearing loss. In the recovery course, the amplitude and threshold of DPOAE is later than that of the pure-tone behavioral threshold, which implies that DPOAE is a sensitive and direct way in reflecting the function of cochlea. PMID- 12541397 TI - [Bilateral large vestibular aqueduct syndrome (7 cases reported)]. AB - OBJECTIVE: To explore the CT features and the auditory threshold in patients with bilateral large vestibular aqueduct syndrome. METHOD: The diameter of vestibular aqueduct in CT and the auditory threshold in each ears of seven patients were measured. RESULT: Asymmetry of the enlarged vestibular aqueduct was found in all patients of whom the side the vestibular aqueduct enlarged was found having a greater hearing loss than the other side in the same patient (paried t-test; P < 0.05). CONCLUSION: The measurement of vestibular aqueduct may be useful in determining the prognosis of patients with large vestibular aqueduct syndrome. PMID- 12541396 TI - [The treatment to esophagus stricture with Ni-Ti shape memory alloy stand(with the report of 19 clinical cases)]. AB - OBJECTIVE: To discuss the use of Ni-Ti shape memory alloy stand in treating the stricture of esophagus. METHOD: After finding the stricture part of esophagus under X-ray, the metal guide string was put through the esophagoscope. Dilatation of esophagus was performed firstly. Then the inserting instrument, which has the Ni-Ti shape memory alloy stand inside, was put through the stricture part along the metal guide string and the stand which can return to its original shape automatically was released. Barium meal examination of esophagus was performed after the operation to determine the site of the stand and to realize the smooth degree of esophagus. RESULT: All 19 patients got a successful operation at one time. They swallowed freely after the operation. The effective rate was 100%. CONCLUSION: Ni-Ti shape memory alloy stand can dilate the stricture of esophagus quickly, improve swallowing function obviously. The operation can be easily performed. The therapeutic effectiveness is reliable, and there is no serious complication. This can be used as a routine method in treating the stricture of esophagus. PMID- 12541398 TI - [The clinical characters of tuberculous otitis media]. AB - OBJECTIVE: To sum up the clinical characters of tuberculous otitis media. METHOD: 2 cases of tuberculous otitis media and other reports are reviewed. RESULT: The onset of this disease is sudden and rapid with little discharge coming from the ear. Sometimes one or more perforation and granulation tissue can be found. Necrotizing bone may be seen by radiography. CONCLUSION: According to the tuberculous history of patients or their families and the clinical features, the diagnosis and the treatment to this disease are not difficult. PMID- 12541399 TI - [Glossus flap for hypopharyngeal reparation]. AB - OBJECTIVE: To research the surgical type for hypopharyngeal reparation. METHOD: The glossus flap with glossus deep arterial was made to repairing hypopharyngeal breach after total laryngotomy and partial hypopharyngotomy in 12 cases of repeated laryngeal carcinoma and hypopharyngeal carcinoma. RESULT: 11 cases recovered in the swallowing function after 15 days and the pharyngeal fistula took place in one case, but healed in 35 days postoperatively. The survival rates in 1 year, 3 years and 5 years were 81.8%, 66.7% and 50% respectively. CONCLUSION: The glossus flap is good material for hypopharyngeal reparation with rich blood supply, few complication and high survival rates. PMID- 12541400 TI - [The curative effect of daphnlin spray on treating rhinitis]. AB - OBJECTIVE: To observe the effect of Daphnlin spray on decongesting rhinocleisis and to search effective drugs for treating rhinitis. METHOD: 105 patients collected from chronic simple rhinitis, chronic hypertrophic rhinitis and perennial allergic rhinitis were treated with daphnlin spray (daphnlin group) or ephedrine spray (ephedrine group). RESULT: The total efficiency of daphnlin group in treating rhinitis above is 90.9% and that of ephedrine group (69.3%) in 1 minute and for ephedrine group is mostly (68.7%) between 1 to 3 minutes. The keeping curative effect time for daphnlin spray is mostly (70.4%) above 5 hours and for ephedrine spray is mostly (71.8%) in 3 hours. After a week, the nasal resistances are difference between daphnlin group and ephedrine group. CONCLUSION: Daphnlin spray is better than ephedrine spray in decongesting rhinocleisis, keeping curative effect and in safety, so daphnli is one kind of effective drugs in treating rhinitis. PMID- 12541401 TI - [Characteristics of outward potassium currents of outer hair cell isolated from the guinea pig cochlea]. AB - OBJECTIVE: To study the normal values and characteristics of potassium currents (Ik) of outer hair cell (OHC) isolated from the guinea pig cochlea. METHOD: With the patch-clamp whole cell recording technique and several supplementary methods, the Ik potassium tail current (Iktail) and reverse potential were recorded under the conditions of the various intra and extracellular solutions and various command voltage stimuli. RESULT: Ik possessed evident voltage and time dependence. The peak current appeared within 20 ms. The average activated potential was approximately -32.7 mV. Ik increased fastestly and greatestly in the voltage range from the activating voltage to 0 mV, and it approached saturation approximately at 40 mV. In normal status, no distinct "rundown phenomenon" of Ik appeared. When 10 mmol/L KCl was put in extracellular solution, Ik would increased. Nevertheless, when K+ was substituted by Cs+ in the intracellular solution, Ik would disappear. The reversal potential, -64.0 mV in average approximately, could be measured by means of Iktail. Furthermore, by means of slope command voltage stimuli, the processes of Ik activation, increment or decrement and saturation could be observed. CONCLUSION: As showing by the above described features, Ik possessed the characteristics of both an inward rectifier and rapid element of a delay rectifier, which would play important role in frequency tuning, mechanic-electrical transduction, maintenance of resting potential and excitement in OHC. PMID- 12541402 TI - [Ciliary neurotrophic factor in the regeneration of facial nerve]. AB - OBJECTIVE: To study the role of CNTF in regeneration of facial nerve. METHOD: SD Rat facial nerves in temporal bones were transected and then combined end to end. In ciliary neurotrophic factor group (n = 30), lesions were covered by gelatin sponge soaked with CNTF. In control group (n = 18), lesions were covered by gelatin sponge soaked with normal saline. 2, 4, 12 weeks after the operation, both CNTF group and SAL group undergone electrophysiology test and histopathology as well as quantity analysis. RESULT: Two weeks later, both CNTF and SAL group failed in eliciting muscular excitement by stimulating facial nerve; T-test showed significant difference between the two groups(P < 0.05) about the average of myelinated axons 4 weeks later, the latency of facial muscular excitement was 7.650 +/- 2.702 ms in CNTF group and 16.080 +/- 0.162 ms in SAL group. The average number of myelinated axons was 1614 +/- 411 in experimental group and 1094 +/- 343 in control group. T-test showed significant difference between them (P < 0.05). 12 weeks later, latency of facial muscle excitement was 2.016 +/- 0.122 ms in CNTF group and 2.060 +/- 0.162 ms in SAL group. The average number of myelinated axons was 1909 +/- 332 in experimental group and 1847 +/- 422 in control group. T-test showed no significant difference between the two groups (P > 0.05). CONCLUSION: CNTF enhance facial nerve regeneration in adult SD rats. PMID- 12541403 TI - [Establishment of animal model for ischemic facial palsy in rat]. AB - OBJECTIVE: In order to establish animal model for ischemic facial nerve palsy in rats and to observe the intrinsic vascular network of facial nerve in the fallopian canal. METHOD: Selective vascular embolization of the external carotid artery was performed in rats. The animals were killed at 1 and 30 days after operation respectively. 10% gelatin solution with fluorescent dye were injected into their left ventricles. The changes intrinsic vascular network were observed under inverted fluorescent microscopy. RESULT: Facial palsy was presented within 5 to 15 min following arterial embolization, the intrinsic capillaries of facial nerve appeared to be thinner and some of them were blocked by microspheres, especially in the labyrinthine segment. Facial palsy almost recovered after 30 days postoperatively. The capillaries returned to be almost normal shape and only few of them were thinner. CONCLUSION: The establishment of animal model for ischemic facial palsy will make contribution to experimental research and clinical treatment for facial palsy. PMID- 12541404 TI - [Expression of EGFR and PCNA, and DNA content in squamous cell carcinoma of larynx]. AB - OBJECTIVE: To investigate the expression of epidermal growth factor (EGF), proliferating cell nuclear antigen (PCNA) and DNA index (DI) in laryngeal carcinoma, to analyse the correlation between these index and the biological characteristics of laryngeal carcinoma and their values of clinical prognosis. METHOD: Immunohistochemical staining was used to detect the expression of EGFR and PCNA in laryngeal cancer and normal tissue, and with MIPS-I image analysis system DNA contents of cancer cell were measured and made out DNA index. RESULT: The positive rate of EGFR in laryngeal carcinoma was 54.8%, and it was negative in all 10 normal laryngeal mucosa specimens (P < 0.01). The expression of EGFR did not correlate with histological grading and 5-years survival rate (P > 0.05), The positive expression of PCNA and DNA contents in the laryngeal carcinoma were increased with the decrease of tumorous differentiation (P < 0.05). With the increasing of PCNA positive expression and DI, the prognosis of the patients were poorer (P < 0.05). CONCLUSION: EGFR may be related to the process of carcinogenesis in laryngeal carcinoma and was used as an early biomarker identifying premalignant lesions which had the greatest risk of carcinogenesis. PCNA and DI were simultaneously detected can be used as the prediction of tumor malignancy and prognosis. PMID- 12541405 TI - [Expression of EGFR in keratosis of laryngeal and the correlation with malignant phenotype]. AB - OBJECTIVE: To evaluate the roles of EGFR expression in laryngeal keratosis. METHOD: The ABC immunohistochemical method was performed to investigate the expression of EGFR in 32 cases of laryngeal keratosis tissue. RESULT: The EGFR indices in light, moderate and serious non-typical hyperplasia were 12.5%, 22.5% and 47.8% respectively. There were significant differences between heavy with light and middle non-typical hyperplasia (P < 0.05). CONCLUSION: The EGFR might play some role in the process of carcinogenesis of laryngeal keratosis, and be as an useful marker for the assessment of the behavior and prognosis in the laryngeal premalignant lesions keratosis. PMID- 12541406 TI - [Laryngeal function preservation in comprehensive therapy of carcinoma of pyriform sinus]. AB - OBJECTIVE: To investigate the sufficiency of comprehensive therapy in the preservation of laryngeal function of carcinoma of pyriform sinus. METHOD: Since 1986, 33 cases of pyriform sinus carcinoma were treated, the clinical stages were as follows: stage I 2 cases, stage II 9 cases, stage III 12, stage IV 10. The induced chemotherapy agents were cisplatin, methotrexate and pingyangmycin. Surgical treatment include local dissection of tumor, partial hypopharynx and partial larynx dissection. All patients were received postoperative radiotherapy. RESULT: Chemotherapy partial response rate was 42.4%, 3 years survive rate was 54.8%, 5 years survive rate was 38.7%. CONCLUSION: The cisplatin centered combined induced chemotherapy was effective in the treatment of pyriform sinus, the laryngeal function preservation surgical treatment of pyriform sinus carcinoma in T1 T2 and selective T3 patients is practicable. Postoperative radiotherapy can improve the local control rate. PMID- 12541407 TI - [Clinical significance of plasma PGE2 levels in pre- and post-operative patients with laryngeal carcinoma]. AB - OBJECTIVE: To study the clinical significance of PGE2 Levels in patients with laryngeal carcinoma. METHOD: Radioimmunoassay was used to evaluate plasma PGE2 levels in 35 patients with laryngeal carcinoma. RESULT: The plasma PGE2 levels in patients with laryngeal carcinoma were significant higher than that of normal subjects (P < 0.05), but there was no correlation with the stage of laryngeal carcinoma (P > 0.05). The plasma PGE2 levels of postoperative patients with laryngeal carcinoma were significant lower than that of preoperative patients (P < 0.05). After operation, the plasma PGE2 levels of recurrent patients were significant higher than that in patients with no recurrence (P < 0.01). CONCLUSION: The determination of PGE2 levels in patient plasma may help the diagnosis and prognostic judgment of laryngeal carcinoma. PMID- 12541408 TI - [16 cases of laryngeal keratosis treated with viaminati]. AB - OBJECTIVE: To investigate the treatment efficient of viaminati on the laryngeal keratosis. METHOD: All 16 cases of laryngeal keratosis took viaminati. RESULT: 15 cases were recovered. 1 case was developed cancer. CONCLUSION: Viaminati is effective in treating laryngeal keratosis and has not obvious side-effect. PMID- 12541409 TI - [Long-term results of modified Majer-Piquet's operation in the treatment of advanced glottic type laryngeal carcinoma]. AB - OBJECTIVE: To investigate the long term results of modified Majer-Piquet's operation in the treatment of advanced glottic type of laryngeal carcinoma. METHOD: A series of 21 cases treated were analysed, of whom, 11 were T3N0M0, 6 were T3N1M0, 4 were T4N0M0. RESULT: 3 years and 5 years survival rate were 100% and 85.7% respectively. Decannulation rate were 95.2%. All the patients could speak after decannulated, and could take food through mouth without inspiration. CONCLUSION: This operation could be used in the treatment of some advanced glottic type laryngeal carcinoma effectively. PMID- 12541410 TI - [Vasomotor rhinitis following endoscopic sinus surgery: 5 cases report]. AB - OBJECTIVE: To investigate the possibility of endoscopic sinus surgery causing vasomotor rhinitis. METHOD: A retrospective study of 300 patients who received endoscopic sinus surgery. 5 cases out of 300 suffered from vasomotor rhinitis 1-3 weeks following endoscopic sinus surgery. Radio frequency cautery of agger nasi mucous was applied to all 5 patients. RESULT: After 4-6 months of follow-up, 4 cases out of 5 with vasomotor rhinitis were effective and 1 improved. CONCLUSION: Endoscopic sinus surgery may be causative factor for vasomotor rhinitis. PMID- 12541411 TI - [Treatment of nasofacial hemangioma with interventional embolization and pingyungmycin injection]. AB - OBJECTIVE: To investigate a new method with interventional embolization and pingyugmycin injection for treating nasofacial Hemangioma. METHOD: The fillets and coil were used for interventional embolization facial artery, internal maxillary artery, temporal superficial artery, and pingyungmycin injected for 4 patients. RESULT: The tumor and vascular beat in this series of 4 patients were disappeared by follow up of one year. There are not nasal obstruction and epistaxis. CONCLUSION: Treatment with interventional embolization and pingyungmycin injection is a safe and effective method for nasofacial hemangioma. PMID- 12541412 TI - [The retrosigmoid approach surgery of trigeminal never]. AB - OBJECTIVE: To explore the effect of retrosigmoid approach surgery of trigeminal never to treat trigeminal neuralgia. METHOD: Twenty-six patients underwent the selective trigeminal neurectomy, among them 2 cases of surgical removal of the cerebellopontine angle cholesteatoma and 2 cases of trigeminal never branch No. I splitting; four patients underwent microvascular decompression. RESULT: By 1-7 years of following up, only one case of microvascular decompression recurrence in all successful operations. CONCLUSION: The effect of retrosigmoid approach surgery of trigeminal never is satisfactory. To know the management of the complications is very important too. PMID- 12541413 TI - [Treatment of mycosis sphenoiditis with endoscopic sinus surgery]. AB - OBJECTIVE: To evaluate the treatment effect of endoscopic sinus surgery for mycosis sphenoiditis. METHOD: From 1995 to 1998, 9 patients of mycosis sphenoiditis were treated with endoscopic sinus surgery. RESULT: In the follow-up of 6-36 months, all the 9 patients were cured. CONCLUSION: Treatment with endoscopic sinus surgery is a safe and effective method for mycosis sphenoiditis. PMID- 12541414 TI - [Diagnosis of nasal sinus fractures and its emergency management: 70 cases report]. AB - OBJECTIVE: To study and improve the diagnosis and treatment level of nasal sinus fractures in the emergency. METHOD: 70 patients suffered from nasal sinus fractures were classified by degree of lesion. Partial of them (24 cases) were treated by Caldwell-Luc method and endoscopic sinus surgery for reducing fractures. RESULT: The results of 22 out of 24 cases were satisfied and the other 2 good. CONCLUSION: To better diagnose, one must pay attention to the symptoms of eyes, orbital floor and nasoethmoid sinus fracture and other severe injuries, CT scan is also necessary for diagnosis. PMID- 12541415 TI - [Immunologic histochemical observation of anti-crypt keratin of cryptic epithelium of tonsilla in children with chronic tonsillitis]. AB - OBJECTIVE: To investigate anti-crypt keratin (CK) immunologic histochemical changes in children with chronic tonsillitis. METHOD: Removed tonsilla were fixed by 10% formaldehyde. Immunologic histochemical method was used to determine the changes of anti-broad spectrum (KD 68, 56, 56, 50) CKSP. RESULT: In 230 cases, obvious keratosis was 90.9%, no keratosis was 9.1%, 3 cases were found with fungus filaments and bacteria in the bottom of crypts. Anti-broad spectrum and hypermolecule CK of tonsil cryptic epithelium were positive reaction, anti-broad spectrum CK of cryptic keratosis in all cases was positive reaction. CONCLUSION: During the period of episode, cryptic epitheliums of tonsilla was destroyed repeatly, therefore, immunoglobulin production was reduce. Because the immune function of tonsilla was reduced, bacteria and virus might be invade into organism. This reduplicative malignant circles must be interrupted or blocked only by tonsillectomy. PMID- 12541416 TI - [An anatomical investigation relating to the stapes elevation surgery]. AB - OBJECTIVE: For the purpose of improving patient's hearing in stapes elevation surgery. METHOD: The anatomical features relating to this operation were observed and measured in 30 ears of fresh cadavers, which included the length of lenticular process, the highness of stapes head and neck, the thickness of lower border of footplate, the angle of incudostapecial joint and the gradients of upper and lower wall of vestibular niche. The relation between two types of the present operation and the investigation were discussed. RESULT: The two types of present operations were in accord with the physiologicoanatomical features of the stapes and incus. CONCLUSION: In order to improve the patient's hearing, making use of biobinder for strengthening the incudostapedial connexion as a routine is suggested. PMID- 12541417 TI - [Traumatic perilymphatic fistula of round and oval windows (four cases reported)]. AB - OBJECTIVE: To explore the diagnosis and treatment to traumatic perilymph fistula (PLF) of round and oval window. METHOD: Traumatic PLF was diagnosed by the traumatic history of head, neck and ear, the examinations of auditory and vestibular function, and the exploratory tympanotomy. The PLF of round and oval windows were repaired by fascia graft or tragus perichondrium and gelform. Four cases with traumatic PLF of round and oval window were reported in this paper, which included 2 cases hitten by hand, 1 by brick, 1 insulted by middle ear surgery. Exploratory tympanotomy was performed from one and half to nine months after injury. One case was misdiagnosed as Meniere's disease before confirming PLF. Exploration and repair of PLF underwent in all cases. RESULT: The symptom of vertigo relieved in all cases after surgery, while the hearing recovery was not evident. CONCLUSION: 1. Traumatic PLF is unrare, hence, traumatic PLF should be alerted if patients suffer from vertigo and hearing impairment after head and ear injury. 2. The features with vertigo attacks, fluctuating hearing loss and tinnutis should be distinguished from Meniere's disease. 3. Early exploratory tympanotomy and repair of PLF are effective for relieving vertigo and improving hearing. PMID- 12541418 TI - [The clinic application of multichannel cochlear implant with eleven cases reported]. AB - OBJECTIVE: To review the effects of profound sensorineural deaf patients who were implanted multichannel cochlear. METHOD: Eleven profound deaf patients (6 prelingually deaf and 5 postlingually deaf) were implanted with twenty-two (model Minisystem 22) and twenty-four (model Nucleus 24) channel cochlear. Mapping of cochlear speech processor was done in 1 to 1.5 months after operation. RESULT: All of 11 patients obtained the hearing. The patients with postlingually deaf can dialog and communicate with telephone after a simple language training without lip reading training. These patients achieved the best results, they not only stepped in sound world but also obtained a normal living and work. In the prelingually deaf patients, they got hold of incoordinate progress in speech ability after assess and speech training. CONCLUSION: The multichannel cochlear implant had good effects in profound deaf patients, especial in postlingually deaf patients and younger children. PMID- 12541419 TI - [The immunohistochemistry and clinic study on surgical margin of laryngeal cancer]. AB - OBJECTIVE: To probe into the safe incisal range of removing laryngectomy cancer determined from the albumen balance of cells. METHOD: LSAB or SABC of immunohistochemistry was applied, and three groups of samples with cancer tissue, nearby cancer tissue and distal cancer tissue of 36 primary laryngectomy cancer were tested by antibodies p16, p21, TGF-beta 1, PCNA and EGFR. RESULT: The difference of the positive rate with the five antibodies between the cancer group and the two side incisal group were very notable with P < 0.01 or P < 0.05, but which between nearby cancer tissue and distal cancer tissue were not notable with P > 0.05. The differences of the positive rates with 5 antibodied between high differentiation group and low differentiation group were notable with P < 0.05, and P < 0.05 with PCNA in the group whether the lymphonodi cervicales transmitted or not. The incisals for all the cases are beyond 5 mm, the recurerence rate is 3%. CONCLUSION: The 5 mm incisal for laryngectomy cancer is appropriate on the albumen cevel of cells. PMID- 12541420 TI - [Clinic and immunophenotypic studies on midline malignant histiocytosis]. AB - OBJECTIVE: To understand the clinic opatholgic changes of midline malignant histiocytosis (MMH), and to explain the source of atypical lymphoid cells (ALC) of MMH, so as to search after the relation between MMH and malignant lymphoma. METHOD: Histopathologic section of 39 cases of MMH were taken HE staining and the histopathologic changes were obsered under microscope. The immunohistochemical staining was also used to examine the immunophenotype of ALC. RESULT: 1. Pathologic changes of MMH: coaglation necrosis and mixed inflammatory cells infiltration, with epithelial infiltration which distributed around the center of blood vessel were their characteristics. 2. Immunohistochemical staining result: 28 in 39 cases of MMH (71.8%) expressed the immunophenvtype of T-cell; among them 13 (33.3%) were finded both positive reaction to CD3 and CD57 antigens with ALCs at the same time. CONCLUSION: According to immunophenotypic studies, most ALCs came from proliferation and infiltration of atypical T cell. ALCs of MMH expressed not only T-cell markers (CD3), but also NK-associated antigen (CD57). MMH was a mucosa related peripheral T/NK cell lymphoma originally. PMID- 12541421 TI - [A treatment to serious esophageal cicatrices stenosi by metal and silica gel dilator]. AB - OBJECTIVE: To find an effective method of treating the esophageal cicatricial stenosis. METHOD: Six cases with esophageal cicatricial stenosis were treated by mental and silica gel dilator. RESULT: The effects in all six cases were satisfactory and no any complications were finded. CONCLUSION: The method is safe, effective and of no complications, the treatment time is shorter also. PMID- 12541422 TI - [The normal parameters in people with static posturography]. AB - OBJECTIVE: To get the normal parameters of static posturography in people. METHOD: 279 normal people from 5 to 80 year old were measured when their eyes opened and closed separately by G-5500 gravity sway apparatus. RESULT: The normal parameters of static posturography were acquired. CONCLUSION: The result can be taken as normal in clinical examination. PMID- 12541423 TI - [The expression of the epidermal growth factor-receptor mRNA in laryngeal squamous cell carcinoma and its correlation with clinical pathological characteristics]. AB - OBJECTIVE: To study the expression of the epidermal growth factor-receptor(EGF-R) mRNA in laryngeal squamous cell carcinoma and its correlation with clinical characteristic and difference of tissues. METHOD: All 20 samples of laryngeal squamous cell carcinoma were determined by dot blot. RESULT: The EGF-R mRNA expression was evaluated in 20 laryngeal squamous cell carcinomas and 15(75%) were finded with EGF-R mRNA expression. When the mRNA of the laryngeal squmous cell carcinoma was decreased, the expression of the EGF-R mRNA was decreased. The EGF-R mRNA expression did not correlated with age or tumor localization (P > 0.05) but correlated with differentiated tumors (P < 0.05) and the stages of TNM according to classification (P < 0.05). Four patients with recuence cancers and six patients with the metastatic cervical-lymph-node involvement had an enhanced expression of EGF-R mRNA. CONCLUSION: The uncontrolled growth and proliferation of the laryngeal squmous cell carcinomas may be related to the abnormal EGF-R mRNA expression. The enhanced expression of EGF-R mRNA was correlated with the differentiation and stages of laryngeal squmous cell carcinomas. PMID- 12541424 TI - [The clinic features of human immunodeficiency virus (HIV) on ear, nose, throat head and neck]. AB - OBJECTIVE: To identify the clinic features of HIV infection on ENT-Head and neck area and to explore its treatment. METHOD: 68 HIV cases in zambia were analysed. RESULT: The ages of the patients are from 3 months to 46 year old. Among them 12 cases with Lump of neck, 9 with ulcer of oral-pharynx, 9 with parctid gland enlarged, 2 with submaxillary plleymon, 5 with nasal bleeding, 17 with purulent otitis Media, 4 with atrophic rhinitis. 18 cases in 58 HIV patients had tubericulosis, diarhea and anemia. 51 cases improved after treatment while 15 cases didn't improve and 2 died. CONCLUSION: It is very important to find and treat HIV patients and to prevent transmission as early as possible. PMID- 12541425 TI - [Experience in endoscopic sinus surgery in district hospital]. AB - OBJECTIVE: To explore the importance and specificity of conducting endoscopic sinus surgery (ESS) in district hospital. METHOD: Seventy patients with chronic nasal sinusitis or/and nasal polyps undergone ESS from June 1997 to October 1998 were retrospectively analyzed. The curative effect was evaluated according as FESS-95 Guangzhou Standard. RESULT: In all patients, 52 cases (74.3%) were cured, 16 cases (22.9%) improved, 2 cases (2.8%) unimproved and 4 cases (5.7%) were found with complications. CONCLUSION: The results showed that intranasal endoscopic sinus surgery had a significant effect on treating nasal-sinusitis and nasal polyps. The CT types of ethmoid sinus are significant in selecting anesthesia and operation program. To get a good result, a full combined therapy in the perioperation period and a careful endoscopic follow-up postoperation are important. PMID- 12541426 TI - [Improvements on radical neck dissection]. AB - OBJECTIVE: To improve the surgery of radical neck dissection for a better curative effect. METHOD: Some improvements on incision, path to exploration and manipulation of operation were made in 65 patients with head and neck tumors while radical neck dissection were done. RESULT: The average surgical time was 110 minutes and the average amount of bleeding was 120 ml in the patients when radical neck dissection had been completed. The complications decreased to 1.54%. The surviual rates of 2 and 3 years are 86.2% (56/65) and 81.6 (31/38) respectively. CONCLUSION: With these improvements on radical neck dissection, the curative effect in the near future is improved. PMID- 12541427 TI - [The investigation of the audio-vestibular and oculomotor functions in people more than eighty years old]. AB - OBJECTIVE: To investigate audio-vestibular and oculomotor functions of octogenarian. METHOD: The audition were measured with audiometry and auditory brainstem response. The semicrcular, oculomotor and balance functions were examinated by electronystagmography and posturography. RESULT: 1. The hearing abnormality of speech frequency were 84.1 percent by audiometry; 2. The abnormal rats of semicircular function and posturogram were 38.5% and 74.4% respectively. The body sway locus and velocity of octogenarians have increased as compared with the group of 60 years old in the posturography; 3. The latency of saccade was prolonged and total hearmonic distortion of pusuit was increased than that of 60 years old in the oculomotor test. CONCLUSION: The easily falling of octogenarians may be related to disturbing of semicircular oculomotor and equilibrational functions. PMID- 12541428 TI - [The rebound of nasal resistance after decongestant used in nose]. AB - OBJECTIVE: To investigate the effect of decongestant on nasal resistance so as to provide theories for studing rhinitis medicamentosa. METHOD: The changes of nasal blockage in 92 cases of acute rhinitis and chronic rhinitis after using decongestants were recorded, and the nasal resistance in 39 cases were measured by active anterior rhinomanometry. RESULT: According to the diseases and decongestants used, the percentage of nasal resistance rebound were 63.5%, 31.2%, 32.2%, 8.6% in 4 groups of investigated patients and 50%, 20%, 75% in 3 groups of measured patients respectively. CONCLUSION: Decongestants can cause the rebound of nasal resistance, which related to the diseases and the decongestants used. The mechanism may be that the factors dilating the vasa in nasal mucosa are activated. PMID- 12541429 TI - [Evaluation of 99mTc(V)-DMSA imaging in patients with pharyngolaryngeal carcinoma]. AB - OBJECTIVE: To evaluate the value of tumor-seeking agent 99mTc(V)-DMSA in patients with pharyngolaryngeal carcinoma and its biodistribution in human being. METHOD: Planar and SPECT scintigraphy with 99mTc(V)-DMSA were performed in 12 patients with histologically confirmed pharyngolaryngeal carcinoma. The dynamic distribution in patients was also analyzed. RESULT: 9 cases (11 lesions) showed increased uptake by planar imaging and 11 cases (19 lesions) were clearly demonstrated by SPECT images. Furthermore, distant metastatic lesions were showed in 5 patients. Biodistribution in patients showed the accumulation of 99mTc(V) DMSA in kidney was significantly greater than other organs. CONCLUSION: These results could suggested the clinical utility of 99mTc(V)-DMSA, specially with SPECT, for detecting primary pharyngolaryngealcarcinoma and their metastatic lesions (both local and distant). 99mTc(V)-DMSA Tumor-seeking agent Pharyngolaryngeal carcinoma. PMID- 12541430 TI - [The significance of the alerting tasks in the vestibular's caloric test]. AB - OBJECTIVE: To evaluate the vestibular's sensitivity to the stimulus of the cold and hot water. METHOD: Forty volunteers are randomly divided into four groups, group one for maths calculation, group two for answering the questions (they are active alerting groups) and group three and four for listening to music and to stories respectively (they are passive alerting groups). Each group has ten volunteers, their left ears are separately stimulated with the cold and hot water, recording the nystagmus slow phase velocity for 20 s at the highest level with the nystagmography instrument of HB-3 ENG computer treatment system. RESULT: There are significant differences between the two active alerting groups and the two passive ones (P < 0.01). CONCLUSION: In performing the vestibular's caloric test, it's more reasonable for clinic to use active alerting tasks. PMID- 12541431 TI - [Hemangioma of external ear: an analysis of 8 cases]. AB - OBJECTIVE: To investigate approaches of diagnosis and therapy to hemangioma of external ear and improve the effects of diagnosis and therapy. METHOD: Eight cases of hemangioma of external ear were analyzed retrospectively. All patients were unilateral hemangioma. 5 were performed digital subtraction angiography (DSA) before operation, of which 3 were performed embolization simultaneously. 7 were operated, 1 was not done. Racemose hemangioma was 5 and cavernous hemangioma was 2 according to pathologic diagnosis. RESULT: All patients have followed up 1 to 5 years, 5 were healed, 2 were controlled, and 1 was not treated. CONCLUSION: Surgery was effective approach to hemangioma of adults. It suggested that it was the key of controlling bleeding during operation that DSA and embolization before operation or ligating artery of blood supply during operation. PMID- 12541432 TI - [Study on stepwise regression of risk factors in patients with sudden deafness]. AB - OBJECTIVE: To explore the affected factors of sudden deafness. METHOD: To analyse the stepwise regression of various factors in age, systolic pressure/diastolic pressure, immunoglobulin G, A, M, KPTT, PT, Fbg, FDP, ELT and auditory threshold. RESULT: There was significant correlation (positive straightline relativity) between the auditory threshold and KPTT in patients with sudden hearing loss, and between the auditory threshold and PT in the analysis of coagulogram. CONCLUSION: The state of blood circulation didn't show hypercoagulability in some patients with sudden hearing loss. We must be very careful in therapy of anticoagulation and the increment of blood volume. PMID- 12541433 TI - [Nonsyndromic deafness and mitochondrial DNA mutation]. AB - OBJECTIVE: To analysis whether there is any mtDNA 1555A-->G homoplasmic point mutation among familial nonsyndromic deafness. METHOD: Polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) were used to screen the mutation 1555A-->G among four nonsyndromic deafness families. RESULT: The same mutations were checked out in 4 of 5 individuals in 1 of 4 families. CONCLUSION: The 1555A-->G change on mtDNA might be one of the multiple genetic defects and pathogenetic of familial nonsyndromic deafness. PMID- 12541434 TI - [Clinical and hereditary study of cup-ear]. AB - OBJECTIVE: To study morphology and genetics of cup-ear. METHOD: The occurrence and pedigree of patients with cup-ear were investigated in Benin, West Africa. Clinical observation and home visit were conducted to see if these patients had hearing loss or other anomalies. RESULT: Between 1996 and 1997, 9 patients with cup-ear were found, accounting for 0.303% of patients seen during this period. These 9 patients were from 7 different tribes. 5 patients had family history, 3 pedigrees were studied in details. CONCLUSION: the family investigation suggested that the formation of cup-ear was determined by the cup-ear gene of an affected parent and it was an autosomal dominant inheritance. PMID- 12541436 TI - [Computed tomographic findings of the development of the bone around the vestibular aqueduct in Meniere's disease]. AB - OBJECTIVE: To study the development of the bone around the vestibular aqueduct (VA) in Meniere's disease (MD) by CT. METHOD: The experiment consisted of three groups; normal ear group, non-MD vertigo group and MD group. VA in each group was examined by CT, and the minimum distance between the posterior semicircular canal and the posterior petrous surface where contain the endolymphatic sac. RESULT: VA visualization rate in MD was low, P-P distance in MD group was shorter than that of normal ear group and non-MD vertigo group. CONCLUSION: It is the fundamental pathological anatomy in MD that VA and the bone around VA are maldeveloped. PMID- 12541435 TI - [Effects of selective attention on distortion product otoacoustic emissions: depend on parameters]. AB - OBJECTIVE: In order to investigate the effects of parameters of selective attention on DPOAEs. METHOD: We designed two kinds of visual tasks. One was two letters O and Q displaying on computer screen. The other was a red-blue light device. Letter Q and blue light were target stimuli flashing at random (probability was 20%), letter O and red light were non-target stimuli occurring orderly (the probability was 80%). DPOAEs with and without visual tasks were tested in 7 normal subjects (14 ears). RESULT: Both of two kinds of visual tasks suppressed DPOAEs and there was no significant difference between them (P > 0.05). Long time test and retests reduced the suppression or even made it disappear. CONCLUSION: It was suggested that parameters influence the suppressive effects. The test should not last over 1 minute and once or twice may be optical. PMID- 12541438 TI - [Clinical analysis of vocal cord leukoplakia in 32 cases]. AB - OBJECTIVE: To investigate the types and curative effects of the leukoplakia of the vocal cord. METHOD: From 1990 to 1996, 32 patients (38 sides) with leukoplakia of vocal cord were operated in our hospital, postoperative follow-up ranged from 2 to 7 years. RESULT: There were eighteen cases of simple leukoplakia and fourteen cases of leukoplakia with mild, moderate and severe atypical hyperplasia in this study. None of simple leukoplakia had malignant change. But there were 33%, 75% and 75% of cases had malignant change in the leukoplakia with mild, moderate and severe atypical hyperplasia respectively. To surgical methods, there was no significant difference, in the rate of malignant change either by the microlaryngoscopy or thyrotomy. CONCLUSION: Simple leukoplakia can be cured. More attention should be paid to the leukoplakia with any types of atypical hyperplasia during clinical management. The postoperative results were not depend on the surgical methods. PMID- 12541437 TI - [Operations for laryngeal carcinoma with reservation of laryngeal function]. AB - OBJECTIVE: To study the long term results and functional recovery of partial laryngectomy in the treatment of glottic and supraglottic types of laryngeal carcinoma. METHOD: A series of 70 cases operated on between July 1978 and August 1998 were analysed, of whom 60 were glottic, 10 were supraglottic type laryngeal carcinoma. Among them, 22 cases underwent vocal cord resection, 22 vertical hemilaryngectomy, 17 Majer-Piquet's operation, 7 horizontal hemilaryngectomy, 2 Arslan's operation. RESULT: 1, 3, 5-year survival rates were 98.53%, 87.04% and 78.26% respectively. Decannulation rate was 100%. All patients could finally take food by mouth without inspiration, and could speak as soon as decannulated. Complication and recurrence rates were 15.71% and 13.04% separately. CONCLUSION: Partial laryngectomy is an effective operation on laryngeal carcinoma with reservation of laryngeal function. It not only can totally excise neoplasm and prolong life, but also can better reserve laryngeal function and improve the quality of patient's life. PMID- 12541439 TI - [Determination of serumal soluble interleukin-2 receptor levels in patients with tonsillitis]. AB - OBJECTIVE: To observe serumal soluble interleukin-2 receptor levels in patients with tonsillitis. METHOD: SIL-2R levels in serum were detected in 68 patients with acute or chronic tonsillitis and 68 normal controls. RESULT: The SIL-2R concentrations in patients with acute tonsillitis and in patients with chronic tonsillitis within attacking period were (642.2 +/- 87.2) x 10(3) U/L and (762.3 +/- 90.5) x 10(3) U/L, respectively. Both were significantly higher than those of the controls (285.5 +/- 49.6) x 10(3) U/L (P < 0.001). Moreover the latter was higher than the former (P < 0.001). There was no difference between the patients with chronic tonsillitis within non-attacking period and the controls (P > 0.05). After treatment of antibiotics for 3-5 days, the SIL-2R Concentrations in serum in valid cases were obviously becoming lower. CONCLUSION: Determination of SIL-2R in serum was valuable for the diagnosis and curative effect judgement of tonsillitis. PMID- 12541440 TI - [The effect of chronic ototoxicity of gentamicin on the function of auditory efferents]. AB - OBJECTIVE: To study the effect of chronic ototoxicity of gentamicin on the function of the cochlea and auditory efferents. METHOD: The effect of chronic ototoxicity of gentamicin on the function of the cochlea was determined by recording thresholds of the compound action potentials (CAP) of the auditory nerves to tone pip at 4, 6, 8, 10 and 12 kHz. The function of the medial olivocochlear (MOC) system was determined, through the observation of the effect of the contralateral noise (CLN) on CAP, before and after gentamicin administration. RESULT: The suppression effects of CLN on CAP were irreversibly eliminated, and the effects were most obvious at the 11th week after gentamicin (P < 0.01), and the response threshold of CAP was raised about 10 dB and 25 dB, respectively, at the 3rd and 11th week after the application of gentamicin. These results were most related to the morphological changes on cochlear efferents and hair cells. CONCLUSION: The auditory efferents play an important role in the chronic ototoxicity of gentamicin. PMID- 12541441 TI - [Morphological observation and electrophysiological properties of isolated Deiters' cells from guinea pig cochlea]. AB - OBJECTIVE: To dissociate viable isolated Deiters' cells from guinea pig cochlea and perform morphological measurements under inverted microscope; To study electrophysiological properties of Deiters' cells. METHOD: Deiters' cells were dissociated from guinea pig cochlea by papain digestion followed by trituration. Standard patch clamp methods were used to measure whole cell capacitance, zero current potential, reversal potential and potassium ion currents of Deiters' cells in normal external solution. Equations were used to compute the relative membrane permeability ratio of Deiters' cell to K+ and Na+ (PK/PNa) and the potassium equilibrium potential (EK). RESULT: Isolated Deiters' cell looks like a comma, which was divided into cell body, stem and Deiters' phalanx and the nucleus is at about center of its body. Cell length of Deiters' cell was (61.6 +/ 20) microM (mean +/- SEM, n = 45), whole cell capacitance was (27.5 +/- 9.2) pF (n = 28), zero current potential was (-20 +/- 1.8) mV(n = 28), reversal potential was (-64.3 +/- 2.9) mV (n = 24), PK/PNa = 25.4:1 and EK = -82.4 mV. Potassium currents of Deiters' cells were recorded in normal external solution. CONCLUSION: The situation of Deiters' cells in cochlea could be inferred according to length of them indirectly. The ion channel of Deiters' cells was mainly K+ selective. Voltage-dependent and outwardly rectifying potassium currents were prominent in Deiters' cells. PMID- 12541442 TI - [Column efficiency prediction of two dimensional chromatography by artificial neural network]. AB - This paper used artificial neural network (ANN) modeling method to study the relationship between the column efficiency and the operating conditions. This method solved the problem that it is not easy to establish a quantitative model between the column efficiency and its main effecting factors using those traditional modeling methods, as the relationship is usually quite complex and non-linear in fact. The varied-pace BP (back-propagation) learning algorithm was adopted, and a three-layer weight-connected ANN model for a typical dual column system was established. The effective plate number representing the column efficiency acted as the output vectors, while the temperature of the pre-column, the temperature of the main column, the pressure difference between the columns and the vent rate acted as the input vectors. Then the model acquired was used to predict column efficiency (characterized by "effective plate number") under different operating conditions. The results showed that the model predicting value was in consistent with the value found. This work proved that ANN modeling method was suitable for the study on the relationship between the column efficiency of two-dimensional column chromatography system and the operating conditions. PMID- 12541443 TI - [Influence of organic modifier on the retention behaviour in soil leaching column chromatography]. AB - The relationship between capacity factors (k') of 55 nonionic compounds and broad methanol volume percentage (psi) of methanol-water eluent in soil leaching column chromatography (SLCC) was systematically investigated. The compounds consist of 11 chlorobenzenes, 14 alkylbenzenes, 22 polyphenyls and polycyclic aromatic hydrocarbons, and 8 pesticides. Reference soil was dry-packed into a stainless steel chromatographic column (10 mm i.d. x 100 mm) by a homemade pressurizing device, and isocratic methanolwater mixture with psi from 0.0 to 0.80 eluated through the column at a flow-rate of 1 mL.min-1. The column was thermostated at (25.0 +/- 0.1) degree C, and chromatographic peak was monitored by an online ultraviolet detector. The results show that both equations, log k' = log k'w + a psi + b psi 2(1) and log k' = log k'w - S psi (2), well fit the retention values. Equation (2) can be used practically due to few experimental data needed and simpler in formula. Explanation is also given for the existence of the carbon (or chlorine) number rule for two classes of homologous series (i.e. methylbenzenes, n-alkylbenzenes) and weak-polar chlorobenzenes in the SLCC process. The slope and intercept of the rule are also well correlated, and both decreases linearly with increasing eluent psi value. PMID- 12541444 TI - [Synthesis of fluorescent derivatization reagent 2-(4-carboxyphenyl) naphthoimidazole and its application in the determination of alcohols by high performance liquid chromatography]. AB - A new sensitive pre-column fluorescent derivatization method, which employs high performance liquid chromatography(HPLC) with fluorometric detection, for the determination of alcohols is described. The alcohols were transformed into the corresponding fluorescent esters by treating with 2-(4-carboxyphenyl) naphthoimidazole in dichloromethane in the presence of 4-piperidinopyridine (PIP) and 1-isopropyl-3-(3-dimethylaminopropyl)-1-carbodiimide perchlorate(IDC) at 80 degrees C for 10 min. The resultant mixture was analyzed by HPLC on a reversed phase column(Zorbax Bp C8, 4.6 mm i.d. x 250 mm, 5 microns) with acetonitrile methanol (volume ratio was 90:10) as mobile phase, and the sample volume was 20 microL. Monitored excitation and emission wavelengths were 345 nm and 485 nm respectively in determination of the alcohol. The calibration curve of cholesterol is linear between 0 and 2.5 g/L with correlation coefficient of 0.9992. The average recovery was 96.15% with RSD of 1.39%. The lowest detectable mass concentration of cholesterol was 1.0 microgram/L (S/N = 3). PMID- 12541445 TI - [Determination of chiral purity of synthetic amino acids by high performance liquid chromatography]. AB - A series of synthetic beta-heterocyclic and beta-naphthyl alanine and aromatic substituted phenylalanine were separated by reversed-phase HPLC as diastereomeric derivatives obtained after derivatization with (1-fluoro-2,4-dinitrophenyl)-5-L valinamide (FDNP-Val-NH2). The separation was performed with gradient elution. The mobile phase A was acetonitrile containing 0.1% trifluoroacetic acid and mobile phase B was 0.1% trifluoroacetic acid. According to the hydrophobicity of the side chain of the amino acids, 6 different linear gradient programs were selected and all gradients completed in 20 minutes. Good resolution was obtained for neutral amino acids. All L-L diastereomer of neutral amino acids (the first letter refers to the configuration of amino acid) were eluted faster than the D-L diastereomer with retention times less than 16 minutes. In the same gradient eluting system, the hydrophobic parameter (pi value) and the position of the substituent in benzene ring of phenylalanine also influence the retention time of diastereomers. Usually the retention time of the phenylalanine with a large pi value was longer than that with a small one. Among the three phenylalanines containing the same substituent, compounds with the substituent in 2-postion gave the shortest retention time. In the case of basic amino acids, the D-L diastereomers eluted prior to the L-L diastereomers and showed rather poor resolution. The chiral purity (ee) of the optical active amino acids (synthetic DL-amino acids after resolution by different enzymes) were calculated with the equation as follows: ee = [[A(L- or D-) - A(D- or L-)]/[A(L-) + A(D-)]] x 100% (L or D-), A: peak area; L-, D-: amino acid diastereomer. PMID- 12541446 TI - [Study of the miscibility of polymer blends by inverse gas chromatography]. AB - An investigation was made on the thermodynamic parameters chi'23 of the polymer system polyvinyl chloride (PVC)/ethylene-vinyl acetate copolymer(EVA) by inverse gas chromatogrphy(IGC), and the miscibility of the blends was studied by using chi'23 as judging foundation, and it was preliminary discussed the relation between chi'23 and composition of the blend, and molecules-chain structure of polymer, and temperature. The influence of the property of probes on chi'23 was also discussed. Results showed thermodynamic miscibility for PVC/EVA blend was accurately and directly indicated by chi'23. Experimental data confirmed the conclusion reported by other studies. It was immiscible for blends of which the acetate content was lower in EVA copolymer. The experimental temperature was favorable for improving miscibility of blends. The medium content in EVA copolymer resulted in partial miscibility for the blend, but the phase-seperation appeared when the experimental temperature was raised. This is in agreement with the previous studies on polymer blends. The apparent interaction parameter was found to be dependent on the chemical nature of the probe, the values of chi'23 were very different, but most experimental data show the same general trend with temperature and composition of blends. PMID- 12541447 TI - [Polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans in C. I. violet 23 dyes and chloranils]. AB - A simple method for the extraction and analysis of polychlorinated dibenzo-p dioxins and polychlorinated dibenzofurans (PCDD/F) in dyes and chloranils has been developed. Five chloranil samples and a dye sample (C.I. Violet 23) were dissolved and dispersed in sulfuric acid, followed by extracting with toluene. After the clean-up by alumina, silica and Florisil columns, the PCDD/F extracts were analysed by high resolution gas chromatograph/high resolution mass spectrometry (HRGC/HRMS). It was found that the dye and its raw material contained octachlorinated dibenzodioxin/furans (OCDD/F) at 1 to 100 micrograms/g level and hexa-, hepta-congeners ng/g levels. The toxic equivalencies (TEQ) of the samples were all beyond the dangerous level. They are found to be one of the important sources of dioxins in textile fabrics. Further investigations were performed to identify their real source. PMID- 12541448 TI - [A novel method for preparation of C18 ester-bond RP-HPLC packing]. AB - RP-HPLC packings have found widespread application in the fields of biology, chemistry and pharmaceutical industry, etc. C18-bonded phase is one of the most important packing. In order to avoid using toxic and unstable chloro-containing silane as derivatization reagent, a new practical and economic procedure had been developed to prepare RP packing. Octadecyl group was bound onto YQG80, 5 microns silica (Tsingdao Oceanic Chemical Factory, particle size d.p. 5 microns) with gamma-glycidoxypropyltrimethoxysilane as coupling agent. Elemental analysis and chromatographic evaluation confirmed the feasibility of the procedure and RP behavior of the packing. After eluting with 500 column volume of mobile phase at different pH (2, 5 and 8), the column still showed good performance (> 64,000 theoretical plates/m) and stability. PMID- 12541449 TI - Study on the retention behaviour of metal-tetraphenyl porphine chelates in reversed-phase high performance liquid chromatography. AB - The retention behaviour of metal chelates of alpha,beta,gamma,delta tetraphenylporphine (TPP) was studied in reversed-phase high performance liquid chromatography (RP-HPLC). On C18 column, with various organic solvents as mobile phases, the retention of the chelates tends to increase in the following order: ZnTPP < TPP < NiTPP < CuTPP. This retention behaviour can get a good explanation from Horvath's "solvophobic theory". With the assumption that the electrostatic field strength of metal chelates can be measured by the ratio of electronegativity vs. ionic radius (EN/ri) of central metal ion, EN/ri was established to be an index for the retention behaviour of metal-TPP chelates. It was found that lnk' is in good linear relationship with the EN/ri value. PMID- 12541450 TI - [The application of liquid chromatography in genetic engineering]. AB - The recent development of the molecular conformation, renaturation and intermediates on therapeutic proteins produced in biotechnology is reviewed. It also includs the quality control and in preparation of the therapeutic proteins in large scale by liquid chromatography in down-stream technology. This review includes 42 references. PMID- 12541451 TI - [Chromatographic analysis of tea polyphenols]. AB - A review is presented about the chromatographic analysis of the polyphenols. Some of the chromatographic methods such as plate chromatography, gas chromatography, reversed-phase high performance liquid chromatography and some new chromatographic methods are introduced. PMID- 12541452 TI - [Chiral separation of glycerin monosulfides and bisulfides with a stationary phase of amylose-tris(3,5-dimethylphenylcarbamate)]. AB - Amylose-tris(3,5-dimethylphenylcarbamate) (ADMPC) and aminopropylated silica gel(APS) were prepared after the reported methods. ADMPC was immobilized on APS from tetrahydrofuran solution with a coating amount of 15%. The chiral stationary phase was packed in a stainless-steel column(150 mm x 4.6 mm i.d.) by slurry method. The column was used for the enantioseparation of 6 glycerin monosulfides and 4 glycerin bisulfides. Mixtures of hexane and 2-propanol were used as mobile phases. The enantiomers of the monosulfides could be separated quite well, while those of the bisulfides could not at all. This phenomenon shows that the ether oxygen atom of the monosulfides plays a key role in the chiral discrimination process. The retention time of the solutes increases significantly as the amount of 2-propanol decreases. This shows the main interaction between the solutes and the chiral stationary phase is hydrogen bonding. A dynamic model is presented to account for the chiral discrimination mechanism. PMID- 12541453 TI - [Selectivity of poly (4-methyl-5-vinylthiazole) stationary phase in reversed phase liquid chromatography]. AB - In this paper, the selectivity of poly(4-methyl-5-vinylthiazole) stationary phase (PMV CA phase) is comprehensively studied in reversed-phase liquid chromatography (RPLC) mode with methanol-water system. The selectivity difference between PMV CA phase and three commonly used RPLC stationary phases (C18, C8 and Phenyl phase) is shown. Four typical species of standards (alkylbenzens, chlorobenzenes, nitrobenzenes and PAHs) are selected for the selectivity study. It is shown that the PMV CA phase has some special selectivities: 1) It has different selectivity from C18 and C8 phase because the PMV CA phase may form diploe interaction, pi electron interaction and hydrogen bonding interaction (with the nitrogen atom and oxygen atom on the phase) with solutes owing to its polar aromatic thiazole ring and polar ester bond etc. on the phase; 2) It has similar property as phenyl phase because both of them can form pi-electron interaction with solutes. But owing to its polar thiazole ring and other polar group on the phase, which may form dipole interaction and hydrogen bonding interaction with solutes, there are still some differences between these two phases. PMID- 12541454 TI - [Determination of tetramethylpyrazine in animal serum and cerebrospinal fluid by high performance liquid chromatography]. AB - A reversed-phase high performance liquid chromatographic (RP-HPLC) method for the determination of the tetramethylpyrazine(TMP) in Chuanxiong extract, the animal(mouse) serum and cerebrospinal fluid has been developed. The TMP was separated on an ODS column Zorbax SB-C18(4.6 mm i.d. x 250 mm, 5 microns) at room temperature and detected by using UV detector at 270 nm. The mobile phase was methanol-water (50:50, V/V) containing 0.2 mmol/L of NH4H2PO4 flowing at a rate of 0.8 mL/min and 20 microL samples were injected. The detection limit of TMP was 1 mg/L and the calibration curve is linear between 5 and 500 mg/L with a correlation coefficient (r) of 0.999. The recovery of TMP ranged 98%-103%. The extract of Chuanxiong and pretreated serum and cerebrospinal fluid sample are stable for a week at room temperature. PMID- 12541455 TI - [Determination of plasma homocysteine by high performance liquid chromatography with fluorescence detection]. AB - This article report a highly sensitive method specific for the determination of homocysteine in plasma by high performance liquid chromatography with fluorescence detection. Half mL plasma with 100 microL 0.11 mol/L sodium borohydride in 50 mmol/L Tris-HCl (pH 9.0) was kept at 30 degrees C for 30 min, and then 0.5 mL 0.5 mol/L perchloric acid was added. After the mixture was kept at room temperature for 10 min and centrifuged at 15,000 r/min for 10 min, 0.5 mL aliquots of the supernatant solution was pipetted into another vial containing 0.1 mL 3 mmol/L Bromobimane in 1.0 mol/L Na2EDTA (pH 7.0) and 0.7 mL of 90 mmol/L ammonium bicarbonate buffer containing 1.43 mol/L Na2EDTA, pH 8.0. The content was mixed, kept at 37 degrees C for 30 min and centrifuged at 15,000 r/min for 10 min. Then 10 microL aliquots of the supernatant solution was injected into a high performance liquid chromatograph with fluorescence detector. The chromatographic conditions were as follows: an ODS column (4.6 mm i.d. x 150 mm, 5 microns), was eluted with a flow rate of 1.0 mL/min. The fluorescence detector was operated at lambda ex 365 nm and lambda em 475 nm. Mobile phase frompump A was 3% methanol containing 0.25% acetic acid. In gradient elution program the methanol from pump B was as follows: 0-8 min, 5%; 8-15 min, 5%-12%; 15-20 min, 12%; 20-30 min, 12% 20%; 30-32 min, 20%; 32-35 min, 20%-100%; 35-40 min, 100%; 40-43 min, 100%-5%; 43 45 min, 5%. The method proved to be linear in the range of 2.5-80.0 mumol/L with a regression coefficient of 0.9988. The minimum detection limit was 0.5 mumol/L, the recoveries were 94.0%-112.0%, and the RSD values were less than 5.6%. PMID- 12541456 TI - [Determination of vitamin D2 included with beta-cyclodextrin complex in "longmu zhuanggu chongji" by HPLC]. AB - In order to enhance the stability of vitamin D2(VD2), the inclusion complex of VD2 with beta-cyclodextrin(beta-CD) was added in "Longmu Zhuanggu Chongji", a medicine of children for the prevention and cure of rickets. The VD2 in beta-CD VD2 inclusion complex was satisfactorily determined by both reversed-phase high performance liquid chromatography(HPLC) and ultraviolet spectrometry(UV). The experimental results from UV were in well consistent with those from HPLC. The accuracy and recovery of HPLC measurement were analyzed, and their relative standard deviation(RSD) were given to be 0.63% and 1.54%, indicating the method is quite correct and reliable, and the linear regression equation is reasonable. The "Longmu Zhuanggu Chongji" containing beta-CD-VD2 inclusion complexes was saponified by a mixture of KOH, vitamin C (VC) and alcohol-water(1:2, V/V) at 70 degrees C, then extracted with petroleum ether. An accurate content of the VD2 product was obtained by HPLC. A simple and rapid HPLC method, which can detect VD2 in the inclusion complex and in "Longmu Zhuanggu Chongji", has been established in this work. PMID- 12541457 TI - [Analysis of 1-(2,6-dichlorophenyl)-2-indolone and related materials by high performance liquid chromatography]. AB - The separation of 1-(2,6-dichlorophenyl)-2-indolone (DCI) and related materials in the synthesis of medical intermediate DCI from 2, 6-dichlorophenol and phenylamine was investigated with high performance liquid chromatography under different chromatographic conditions. The results showed that DCI and related materials were in baseline separation by using CLC-CN(150 mm x 6.0 mm i.d., 7 microns) as column, MeOH-H2O(6:4, V/V) as mobile phase and a flow-rate at 1 mL/min. A rapid, accurate and reproducible HPLC method for determining DCI was developed. Fine and crude DCI samples were detected, results showed that N-(2,6 dichlorodiphenyl)-chloracetyl amide (CBCC) was the main impurity in fine sample, and CBCC and 2,6-dichlorodiphenylamine were the main impurities in the crude one. PMID- 12541458 TI - [A rapid, isocratic method for analysis of phospholipids by reversed-phase high performance liquid chromatography]. AB - Linear calibration curves were obtained for 4.5-10.5 micrograms of phosphatidylcholine(PC), 1.5-3.5 micrograms of phosphatidic acid(PA), 9.0-21.0 micrograms of cardiolipin(CL), 2.25-5.25 micrograms of phosphatidylglycerol(PG) by reversed-phase high performance liquid chromatography with UV detector at 205 nm, P-E C18 column (15 cm x 0.46 cm i.d., 5 microns) and isocratic elution with mobile phase of methanol-acetonitrile-water (79:8:13, V/V/V) at a flow rate of 2.0 mL/min. Separation and detection of these four standards were achieved reproducibly and easily in less than 12 minutes. The RSDs of peak area measurements were from 0.77% to 8.86%. Average recovery of PC was 89.21% (RSD = 2.1%). The air-dried bear biles were ground into powder and extracted continuously with chloroform-methanol (1:1, V/V) to a final dilution 25 folds the volume of the sample. The extract was washed by addition of half its volume of 0.58% aqueous NaCl. The resulting mixture separated into two phases. The lower phase was the total pure lipid extract and was dried at room temperature under nitrogen. The lipid residue was dissolved in methanol to the desired volume and injected into the HPLC system for analysis. The results indicated that six kinds of bear biles had differences in the phospholipid composition. The method enables a simple, rapid and reproducible quantification of PC, PA, PG and CL. PMID- 12541459 TI - [Determination of major components in Psoralea corylifolia L. by using supercritical fluid extraction (SFE)]. AB - A method based on SFE and CGC has been developed for qualitative and quantitative analysis of major components in Psoralea corylifolia L.. The orthogonal design method and the analysis of variance are used to optimize the five variables (temperature, pressure, volume of modifier, static extraction time and CO2 dynamic extracting volume) of SFE and the optimized conditions listed as follows: temperature 70 degrees C, pressure 38.5 MPa, static extraction 1 min, dynamic extraction 7 mL and 60 microL CHCl3 as modifier. The recoveries of psoralen and isopsoralen are 96.93% (RSD 1.87%) and 98.43% (RSD 2.06%) respectively. Being compared with conventional methods, the SFE as an alternative technique for sample preparation will be most useful in the quality control of Chinese traditional medicines. PMID- 12541460 TI - [Determination of dicofol residue in tea by wide-bore capillary gas chromatographic column]. AB - Dicofol residue is harmful to health. More and more countries have established the limitation of dicofol in foods. This paper describes an efficient method of determination for the dicofol residue in tea. The dicofol was extracted from the tea sample with 20% acetone-hexane, cleaned up on a column of Florisil and acidic siliceous earth (sulfuric acid 0.3 mL/g) in series. Then the column was washed with 10 mL, 20% dichloromethane-hexane, the flow rate was 1 mL/min. At last dicofol was hydrolyzed with potassium hydroxide solution, forming p,p' dichlorobenzophenone(DBP), which was separated from other ingredients through wide-bore capillary(LZ-II, 25 m x 0.53 mm i.d.) and determinated by gas chromatography with electron capture detector(ECD), using Aldrin as internal standard. When the mass ratio of dicofol was in the range of 0.05-3.0 mg/kg, the recoveries were 78%-104% and the limit of determination was 0.5 microgram/kg. This method is simple, sensitive and suitable for pesticide residue analysis. It can also be applied to the determination of dicofol residues in other plant samples such as vegetables, fruits and so on. PMID- 12541461 TI - [Separation and determination of three plant internal hormones by gas chromatography]. AB - A method for the simultaneous separation and determination of three plant internal hormones indole acetic acid (IAA), abscisic acid(ABA) and gibberellic acid(GA) in plant by gas chromatography with wide-bore capillary column is described. The gas chromatographic conditions were as follows: FID detector; HP-1 capillary column, 5 m x 0.53 mm x 2.65 microns film; the column temperature, 220 degrees C; the injector temperature, 250 degrees C; the detector temperature, 280 degrees C, carrier gas, 3.5 mL/min N2; internal standard n-docosane. All components and internal standard were separated in 8 min. The detection limit of IAA, ABA and GA were 0.16, 0.08 and 0.48 mg/L respectively. The relative standard deviations were 2.2%, 1.7% and 2.8% respectively. The linear range were 0.16-80 mg/L(r = 0.9986), 0.08-40 mg/L (r = 0.9993) and 0.48-240 mg/L (r = 0.9991) respectively. The average recoveries were (88.4 +/- 2.4%)%, (92.2 +/- 1.2)% and (91.8 +/- 1.8)% respectively. The method is simple, rapid, sensitive and reproducible. PMID- 12541462 TI - [Chiral separation of naproxen and flurbiprofen by capillary electrophoresis]. AB - The acidic chiral drugs naproxen and flurbiprofen were successfully separated into two enantiomers when beta-cyclodextrins (beta-CDs) were used as chiral selectors by capillary zone electrophoresis, under the conditions of 0.1 mol/L phosphate buffer with pH 4.92. The comparison of four CDs, namely beta-CD, DM beta-CD, HP-beta-CD and TM-beta-CD for chiral separation was made. Naproxen can be separated by either beta-CD or its derivatives, while flurbiprofen can only be separated by TM-beta-CD among the CDs. The elution order of enantiomers of naproxen in different CDs was also studied, and the R form always eluted before S form when any of the four CDs was used as chiral selectors. The method of chiral separation for weak acidic compounds was also developed. It was proved that the optimum pH value for their chiral separation was about 5, close to its pKa value. PMID- 12541463 TI - [Separation and determination of nycodenz and its impurities by capillary electrophoresis]. AB - Separation and determination of nycodenz (a new radiography reagent) and its impurities by capillary electrophoresis was investigated. Since the analytes were polyols, Na2B4O7-H3BO3 was chosen as the buffer system. The analytes were negatively charged by complexing with borate. Then cetyltrimethylammonium bromide was used to reverse electroosmotic mobility in order to shorten the analysis time. 2-Propanol was added to optimize the separation further. Acceptable results of linearity, precision of corrected peak area and migration time were obtained. The RSD(n = 6) of corrected peak area and migration time were 0.73% and 1.4% respectively. The results obtained by CE were directly compared to those obtained by high performance liquid chromatography. PMID- 12541464 TI - [Determination of benzene, toluene and xylene in marine sediment samples by micellar electrokinetic capillary chromatography]. AB - The analysis and determination of the contents of benzene series compounds in marine sediment are important in chemical exploration of petroleum resources. In the present study a method has been developed for this purpose by using micellar electrokinetic capillary chromatography (MECC). A fused silica capillary of 45 cm x 75 microns i.d. (effective length 40 cm) was used. Running buffer was 2 mmol/L aqueous solution of sodium tetraborate containing 75 mmol/L sodium dodecyl sulfate (pH 9.16) and 20% methanol. The applied voltage was 25 kV and detection wavelength was 200 nm. The benzene, toluene and xylene were separated well within 20 min. Peak area was used for quantitative analysis. The calibration curves were linear in the range 4-50 mg/L. The relative standard deviation was within 6.2%. The contents of benzene series in sediments were between 3.79 and 17.36 micrograms/g. PMID- 12541465 TI - [Study on the determination of peptide mixture by HPCE]. AB - A rapid method for the simultaneous assay of 7 peptide mixture, including angiotensin I, II, III, substance P, neurokinin, somatostatin and neurotensin, by high performance capillary electrophoresis has been established. The nature, pH and concentration of buffer, running voltage, detection wavelength, injection time and the effective length of amino-coated capillary were defined with the results of experiment. With 50 mmol/L ammonium acetate (pH 4.5) as running buffer and siphonage injection for 10 seconds, the measurements were carried out at 25 degrees C and 10 kV running voltage [(-)-->(+)] applied to a 57 cm x 75 microns i.d. (50 cm effective length) amino-coated capillary. The 7 peptide mixture was determined by a UV detector at 214 nm. The total time for separation and determination was within 8 min. The recoveries ranged from 95% to 98% with RSD from 2.9% to 4.2%. It has been found that the 75 microns i.d. capillary has higher sensitivity than 50 microns, but its efficiency and Rs were worse. PMID- 12541466 TI - [Determination of aflatoxins in peanut by high performance liquid chromatography using immunoaffinity column clean-up and on-line electrochemical derivatization]. AB - A method for the determination of aflatoxin B1, B2, G1 and G2 in peanut by high performance liquid chromatography using immunoaffinity column clean-up and on line electrochemical derivatization was developed. Sample was extracted with 80% methanol and the extract was passed through the IAC. Aflatoxins were retained by the monoclonal in the IAC and eluted with actonitrile. AFT B1 and G1 were derivatized with on-line electrochemical equipment Kobra Cell. The four toxins can be separated in 13 minutes and the detection limit was 0.1 microgram/kg for each toxin. The method has been applied to the determination of peanut sample. The relative standard deviation was 9.2%-15% and the average recovery of AFT spiked at 0.5-9.0 micrograms/kg levels was in the range of 74.8%-97.3%. PMID- 12541467 TI - [Determination of glucosinolates in rapeseed by reversed-phase ion-pair liquid chromatography]. AB - This paper presents a method for the direct determination of intact glucosinolates in rapeseed by reversed-phase ion-pair liquid chromatography (RILC). The catechin, mercaptoethanol and phytic acid was adopted respectively in sample pretreatment to prevent indole glucosinolates being oxidized. Among them, the effect of mercaptoethanol was the most obvious. The effects of composition and concentration of the mobile phase, the pH of the mobile phase and the column temperature on the retention and the capacity factor of the glucosinolates were studied. The condition of this method by RILC has been set up: column, YWG-C18 H37(10 microns, 250 mm x 4 mm i.d.); mobile phase, 0.02 mol/L KH2PO4 buffer(pH 6) containing 4.5 mmol/L (C4H8)4NBr and CH3CN(90/10, V/V); flow rate, 1 mL/min; detector, UV-226 nm; column temperation, 30 degrees C. In this condition, six glucosinolates were separated completely. The relative correction factors were determined by using sinigrin or benzoic acid as the internal standard. The characteristics of glucosinolates in different kinds of Chinese rapeseed can be determined by this RILC method. PMID- 12541468 TI - [Derivative gas chromatographic analysis of fructooligosaccharide in fermented sucrose]. AB - As a new sweetener, fructooligosaccharide is paid more and more attention for its health improvement property. It includes trisaccharide, tetrasaccharide and pentasaccharide, and can be produced from sucrose by the fermentation of microorganism. In order to analyze the content of fructooligosaccharide in fermented sugar by gas chromatography, fructooligosaccharide was transformed into trimethylsilyl derivatives. Based on the modified gas chromatograph SP2308, and under the chosen chromatographic conditions with 0.53 mm capillary column of OV 101, the contents of fructose, glucose, sucrose and fructooligosaccharide were determined by programmed temperature chromatography. The recovery of fructooligosaccharide was satisfactory. PMID- 12541469 TI - [Trouble shooting of splitter apparatus of Shimadzu GC-14A gas chromatograph]. AB - This paper describes the abnormal phenomena caused by almost complete block of connective tube of SPL-14 split/splitless sample injector of Shimadzu GC-14A gas chromatograph. It has been pointed out that if oil compounds with high boiling point are analyzed, periodical clean-up should be done to remove blocking materials for experimental repeatability and accuracy. PMID- 12541470 TI - [Determination of phenols in environmental water with bromination derivatization and gas chromatography]. AB - A method for the determination of phenols in environmental water with bromination derivatization and gas chromatography is described. The phenols were transformed to bromophenols and then extracted with toluene and separated by the FFAP wide bore capillary column and detected by electron capture detector (ECD). The results showed that the linear ranges were 0.06-80 micrograms/L for o methylphenol and p-methylphenol, 0.03-40 micrograms/L for phenol and m methylphenol. The detection limits for o-methylphenol, p-methylphenol, phenol and m-methylphenol were 0.03, 0.06, 0.01, 0.015 microgram/L respectively. The method is simple, rapid and reliable for determining phenols in environmental water. PMID- 12541471 TI - [The determination of activated partial thromboplastin time, coagulate time and thrombin time in patients with epistaxis of indeterminate cause]. AB - OBJECTIVE: To explore the coagulation mechanism of indeterminate epistaxis. METHOD: 36 cases with epistaxis of indeterminate cause were studies by mean of detecting activated partial thromboplastin time (APTT), coagulate time (CT) and thrombin time. The results of first APTT, APTT after 8 min and CT were observed. RESULT: 1. The first APTT in epistaxis and normal group didn't show statistical difference (P > 0.05). But the APTT after 8 min in epistaxis were significantly different compared with the first APTT and that in normal group (P < 0.01). 2. The CT prolonged in 33.3% patients with epistaxis, which was higher significantly than that in the normal group (P < 0.01). (3) The epistaxis thrombin time (TT) was longer than that in normal group. CONCLUSION: The result suggest that during blood coagulation in indeterminate epistaxis, the activated factors in internal coagulating system may decompose more quickly than that in normal group. Antiagglutinating factors increase in blood. PMID- 12541472 TI - [Nasopharyngeal stenosis following uvulopalatopharyngoplasty]. AB - OBJECTIVE: To report nasopharyngeal stenosis following UPPP and our treatment experience. METHOD: Two clinical cases were analysised. RESULT: In this paper, the complications were attributed to excessive excision palapharyngeal arches, which juried mucous membrane of nasopharyngeal inlet during the long management time as bleeding and the laser used. Surgical management was performed in two cases. One failed. One was treated successfully by rotation palato-pharyngeal flap and prolonged nasopharyngeal stent, Follow-up 20 months there was no recurrence of stenosis. CONCLUSION: Surgical correction of nasopharyngeal stenosis following UPPP is challenging and the best treatment is prevention. PMID- 12541473 TI - [Study on the relationship between allergic rhinitis and airway hyperresponsiveness]. AB - OBJECTIVE: In order to study the relationship between allergic rhinitis and airway hyperresponsiveness. METHOD: Each of them were separately engaged skin test with antigen. All 50 patients had intense positive reaction to artemisia pollen (++-+++), but all controls were negative. We conducted bronchial provocation test (BPT) with 1:100 (w/v) artemisia pollen. Before and after BPT, we separately tested the forced expiratory volume in first second (FEV1) which represented ventilative function and eosinophil cationic protein (ECP) in their serum. RESULT: Before BPT, 50 patients and 20 normal controls had no trachea symptoms. FEV1 of 50 patients is (70 +/- 20)%, that normal controls is (85.6 +/- 8.9)% (P < 0.001). After BPT, in 50 patients, 23 cases felt constriction in the chest, 10 cases had wheezes, 5 cases had asthma, 12 cases had no symptoms. At the same time, all of normal controls had no symptoms. After BPT, FEV1 of 50 patients is (60 +/- 20)%, that of normal controls was (85.6 +/- 8.9)% (P < 0.05), ECP of patients was (30.37 +/- 18.92) micrograms/L, that of controls was (5.245 +/- 3.228) micrograms/L (P < 0.05). CONCLUSION: Patients with allergic rhinitis not only have chronic inflammation in their nose mucosa that results from allergic react, but also have chronic inflammation in bronchus which leads to AHR. The results reveal that most patients with allergic rhinitis have AHR (76%). PMID- 12541474 TI - [The immunologic study on local nasal immunotherapy for perennial allergic rhinitis]. AB - OBJECTIVE: To investigate the clinical efficacy and immune mechanism of local nasal immunotherapy (LNIT). METHOD: 142 cases of perennial allergic rhinitis (PAR) were treated with LNIT. Immunologic study was carried out in 30 cases. RESULT: LNIT was as effective as traditional subcutaneous immunotherapy (P > 0.05). Immunologic study demonstrated, that the nasal secretory IgA (S-IgA) increased significantly after treatment (P < 0.05). The levels of nasal secretory total IgE, serum total IgE and serum specific IgE (anti-mite, ragweed pollon) decreased, but there was no significance (P > 0.05). CONCLUSION: LNIT can improve the local immune function and modify the local nasal allergic inflammatory response. It is an effective, safe and convenient treatment. PMID- 12541475 TI - [Influence of smoking on nasal airway resistance]. AB - OBJECTIVE: To discuss the influence of smoking on nasal airway resistance (NAR). METHOD: The unilateral and total NAR of 40 normal controls, 41 slight smokers and 42 heavy smokers were evaluated by anterior rhinomanometry before and after nasal decongestion. RESULT: Before nasal decongestion, the unilateral and total NAR of heavy smokers increased significantly (P < 0.05) compared with that of normal controls. After decongestion, there was no significant difference (P > 0.05). Whenever before or after decongestion, there was no significant difference between slight smokers and normal controls (P > 0.1). CONCLUSION: Long-time and heavy smoking can enhance the NAR, which may be due to the chang of blood vessels in nasal mucosa. PMID- 12541476 TI - [CT diagnosis of non-Hodgkin's lymphoma in nasal cavity and paranasal sinuses]. AB - OBJECTIVE: To explore the CT characters of non-Hodgkin's lymphoma initially located in nasal cavity and paranasal sinuses. METHOD: The CT manifestations of 9 non-Hodgkin's lymphoma initially located in nasal cavity and paranasal sinuses verified by pathology were analysed retrospectively. RESULT: 1) Non-Hodgkin's Lymphoma of nasal cavity and paranasal sinuses is essentially malignant, acts as a typical malignancy in its clinical manifestation, but on CT, its bone destruction acts as a benign tumor consisting of bone remodeling. 2) Hypertrophic mucosa lies on septum chiefly. 3) Mass with soft tissue density always involves in inferior meatus and inferior turbinate. CONCLUSION: The CT manifestation of non-Hodgkin's lymphoma initially located in nasal cavity and paranasal sinuses has its own characters. PMID- 12541477 TI - [The significance and expression of vascular endothelial growth factor on nasal inverted papillomas]. AB - OBJECTIVE: To determine the expression of vascular endothelial growth factor (VEGF) and its receptor in nasal inverted papillomas (NIP) and to clarify the function of VEGF in the establishment of NIPs. METHOD: VEGF and its receptor, fetal liver kinase-1 (flk-1) expression were examined by immunohistochemistry using SP method in sections of NIPs from 11 patients and inferior turbinates from 6 patients with chronic simple rhinitis. An automatic image analyzer was used to detect the staining results. Gray scale, a half-quantitative parameter, was used to describe the expression of VEGF and flk-1. RESULT: It showed that all the epithelium in NIPs, together with vascular and stroma adjacent to the epithelium, expressed different degree of VEGF. The VEGF expression in epithelium was significantly stronger in NIPs than that in inferior turbinates (P < 0.001). The expressions of both VEGF and flk-1 in vascular endothelium were more intense in NIPs than that in inferior turbinates (P < 0.001, P < 0.001). Flk-1 positive expression was observed in epithelium of NIPs but not in inferior turbinates. CONCLUSION: The results suggest that VEGF participate in the growth of NIPs, part of which shows the characteristics of containing plenty of blood vessels and being bloody in operation. The enhanced expression of VEGF may also play a key role in the development of edematous stroma. The expression degree of VEGF in the epithelium may be identified as one of the parameters of judging the propensity of NIPs' malignant transformation. PMID- 12541478 TI - [Study on the relationship between nitric oxide and allergic rhinitis]. AB - OBJECTIVE: To investigate the relationship between nitric oxide (NO) and allergic rhinitis (AR). METHOD: NO in the serum of 50 patients with AR and 40 healty control were measured by nitrate reductase. RESULT: The level of serum NO in AR group was higher than that in healthy control group (P < 0.001). CONCLUSION: There is a relationship between the level of NO in serum and AR. The results suggest that the NO may play an important role in the pathogenesis of AR. PMID- 12541479 TI - [The clinical features of laryngeal malignant lymphoma with 5 cases reported]. AB - OBJECTIVE: To improve the diagnosis and treatment of laryngeal malignant lymphoma. METHOD: Clinical features of 5 cases were reported and recent literatures were reviewed. RESULT: 2 cases presented with large ulcerated lesions and their laryngeal structure were destroyed, their prognosis were poor. Another 3 cases were local primary lesions, they were generally associated with a relatively short history of hoarseness and dysphagia and exhibited grey-white neoplasms with preferential involvement in the supraglottic parts of larynx, but non-ulcerated lesions, they were treated with operation and radiotherapy or chemotherapy or combinations. The prognosis were very well after 2 to 6 years follow up. CONCLUSION: It is important for the local primary lesion to be diagnosed correctly for the treatment and prognosis. PMID- 12541480 TI - [Combined therapy for 121 cases of advanced cancer of the maxillary sinus]. AB - OBJECTIVE: To investigate the possibility of combined therapy (chemotherapy + surgery) for advanced cancer of maxillary sinus. METHOD: Pre-operative radiation + surgery for 61 cases and preoperative chemotherapy + operation for 60 cases were performed. Radiotherapy referred to the use of linear accelerator and/or 60 Co with the dosage of 40-60 Gy. Chemotherapy referred to the administration of CDDP. VCR, and pingyangmycin. RESULT: The survival rate observed in the 61 cases treated with radiation operation was 36.1%; while the other 60 cases treated by chemotherapy + operation had a survival rate of 31.1%. CONCLUSION: The better way to treat advanced cancer of the maxillary sinus is pre-operative chemotherapy + surgery. In addition, repeated application of chemotherapy can raise the effect of surgical treatment. PMID- 12541481 TI - [Parapharyngeal space neoplasms]. AB - OBJECTIVE: To explore the diagnose and management of parapharyngeal space (PPS) neoplasms. METHOD: Records of 48 patients with PPS neoplasms operated from 1988 to 1998 were reviewed. RESULT: Neurogenic tumors were the most common neoplasma in primary PPS neoplasms. CT and MRI could provide clinically informations such as their size, shape, extent and site. Tumor vessels could be clearly seen by carotid arteriography. The majority of PPS tumors were resected by a transcervical approach. CONCLUSION: CT and/or MRI are essential for diagnose and presurgical planning, Arteriography can be used only when CT or MRi suggest a glomous tumor, or possible involvement of carotid artery, or when those tumors put the carotid artery at a risk for surgical injury. Transoral approach may be used for small tumors and transparotid approach may be especially indicated for removing deep lobe parotid tumors with extention to the PPS. When there is a significant risk for injury the internal carotid artery radiological evidence are necessary for infratemporal approach. PMID- 12541482 TI - [Detection of EBV-DNA, EBNA2 and LMP-1 in tumor tissues of han and uygur patients with nasopharyngeal carcinoma in Xinjiang]. AB - OBJECTIVE: To investigate the possible differences of human nasopharyngeal carcinogenesis between Han and Uygur patients in xinjiang. METHOD: Detection of EBV-DNA, EBNA2 and LMP-1 in tumor tissues of 73 patients (Han 41, uygur 32) with nasopharyngeal carcinoma (NPC) were performed by PCR and immunohistochemistry. RESULT: Tho positive rate of EBV-DNA, EBNA2 expression was 48.2%(20/41), 43.9% (18/41) in Han and 59.3%(19/32), 43.7%(14/32) in Uygur respectively, no significant difference was found (P > 0.05). Expression of LMP-1 in NPC, in both positive and negative EBV-DNA specimens, was higher in Uygur (78.9%) than that in Han (40%) (P < 0.05), CONCLUSION: The EBV infection may involve in nasopharyngeal carcinogenesis in xinjiang; the LMP-1 of EBV could be a more critical factor in malignant transformation of normal nasopharyngeal epithelium in uygur people than in han Chinese. PMID- 12541483 TI - [Detection of distortion-product otoacoustic emissions in well-drilling workers]. AB - OBJECTIVE: To understand the value of using distortion product of otoacoustic emissions(DPOAE) in monitoring noise-induced hearing-loss. METHOD: The amplitude and the positive rates of DPOAE were detected in 102 well-drilling workers (195 ears). RESULT: The amplitude and the positive rates of DPOAE declined significantly in noise groups against control. With the increase of working-age the positive rates of DPOAE was decrease. The DPOAE amplitudes and the positive rates of noise group with normal pure tone threshold were obvious lower than that in controls. There were negative relationship between pure tone threshold in high frequency and the positive rates. CONCLUSION: The DPOAE can reflect the cochlea's function of labors working in noise condition well. Contrast with the pure tone audiometry, DPOAE was more sensitive and more objective. PMID- 12541484 TI - [The influence of serotonin on cochlear microcirculation]. AB - OBJECTIVE: The present study was undertaken to investigate the influence of serotonin (5-HT) on cochlear microcirculation. METHOD: 2 microliters 0.5% 5-HT were applied to round window membrane in anaesthetized guinea pigs. The three parameters as blood flow (CBF), number of blood cell moving (CMBC) and blood velocity (Veloc) of cochlear microcirculation were measured with laser doppler flowmeter. The mean arterial pressure (MABP) was recorded simultaneously. RESULT: 5-HT induced a biphasic response of reduction in cochlear blood flow. In the first phase, cochlear microcirculation showed a transient abruptly reduction. The CBF, CMBC and Veloc reached to 86.8%, 90.2% and 96.3% of the baseline respectively. The second phase reduced slowly. The CBF and CMBC decreased to 89.7% and 89.6% by average throughout the experiment period respectively. The Veloc and MABP had no significant changes. CONCLUSION: 5-HT could decrease the cochlear, blood flow and result in cochlear microcirculation dysfunction. PMID- 12541485 TI - [Neurootologic study of acoustic neuroma]. AB - OBJECTIVE: To summarize the outcome of neurootologic examination of acoustic neuroma and study of diagnostic significance of neurootologic examination on acoustic neuroma. METHOD: 13 patients were examined with pure tune test, acoustic immittance, electronystagmography, acoustic brainstem evoked response, electrocochleogram, otoacoustic emissions and facial electroneurography. RESULT: All patients had subjective hearing loss. One patient had normal pure tone audiogram, other 4 patients had normal pure tone test at the frequency < 2000 Hz. The outcome of acoustic immittance is various. 5 patients were normal in caloric test, but all were abnormal in visual-vestibular optokinetic response. ABR are positive in 90.9% of cases, but 2 patients had no response. 7 patients were abnormal in facial electroneurography, 3 (of 3) eletrocochleogram and 2 (of 2) otoacoustic emissions revealed retrocochlear pathology. CONCLUSION: Neurootologic examination played important role in filtering diagnosis of acoustic neuroma, results should be considered synthetically, acoustic neuroma should not be ruled out if any one test was negative. PMID- 12541486 TI - [The effects of the plasty of the cavity of auricular concha in the post mastoidectomy]. AB - OBJECTIVE: The purpose of this study was to compare the results of mastoidectomy with the plasty of the cavity of auricular concha and the counterpart of mastoidectomy without it at after for 2-6 years followup. METHOD: A retrospective analysis of 320 cholesteatoma cases, surgically treated at the department of otorhinolaryngology-head and neck surgery, Renji hospital affiliated to Shanghai No. 2 Medical University, over a 9-year period, is reported. RESULT: A satisfactory results from the follow-up implicated that this procedure was important for epithelization of the mastoid cavity and the attaining of dry ear after mastoidectomy. CONCLUSION: The plasty of the cavity of auricular concha has an emphasized role in the post-mastoidectomy and is clinically worthy to be applicated. PMID- 12541487 TI - [The clinical significance of analysis of frequency domain of transient-evoked otoacoustic emissions]. AB - OBJECTIVE: To determine the clinical value of analysis of frequency domain of transient-evoked otoacoustic emissions (TEOAE). METHOD: Frequency domain of TEOAE were measured in 1. normal functional ears; 2. noise exposure ears; 3. Meniere's ears; 4. pathologic ears of media ear; 5. contralateral suppression of normal ears. RESULT: 1. After exposure to noise, the value of band signal/noise ratio were decreased in certain frequency band, main frequency of FFT response were changed in frequency spectrum. 2. In the ears of Meniere's disease, after the administration of glycerol, the value of band signal/noise ratio were increased in certain frequency band, no TEOAE were elicited before the administration of glycerol, but they appeared after glycerol administration in some ears, main frequency were changed in frequency spectrum, 3. In the pathologic ears of media ears. The value of band signal/noise ratio decreased to different extent with different kind of pathologic changes, which is related to the change of acoustic immittance. Besides. Acoustic immittance were normal in those with mild pathologic change in the tympanic membrane. The value were decreased in band signal/noise ratio. 4. After contralateral suppression of normal ears, 1 kHz and 2 kHz band signal/noise ratio value were decreased. CONCLUSION: Frequency domain of TEOAE is a sensitive parameter and has property of frequency specificity. It is of great help to study the function of outer hair cells farther and make a differential diagnose in the clinical practice. PMID- 12541488 TI - [Measuring and analyzing of otoacoustic emission tests of a family with genetic progressive sensorineural hearing loss]. AB - OBJECTIVE: To study the diagnostic significance of otoacoustic emission in patients with genetic progressive sensorineural hearing loss. METHOD: To analyse the cochlear function pure tone audiometry Transient Evoked Otoacoustic Emission (TEOAE) and distortion Products Otoacoustic Emission (DPOAE) were measured in 54 members of a six-generation family with genetic progressive sensorineural hearing loss (108 cases) and 50 normal people. RESULT: The family was autosomal dominant heritage. In the group with pure tone thresholds more than 20 dB HL, the amplitudes of TEOAE descended or disappeared in 84% cases and the amplitudes of DPOAE declined or vanished in 82% cases. In the group with normal pure tone thresholds, the amplitudes of TEOAE descended in 75% cases and the amplitudes at one of the frequencies of DPOAE dropped or disappeared in 64% cases. But the incidence of TEOAE and DPOAE in normal people was 100%. CONCLUSION: OAE had important significance in the diagnosis of genetic hearing loss at early stage. PMID- 12541489 TI - [A new method to evaluate the eustachian tube function]. AB - OBJECTIVE: To study the practicality of a new method to evaluate eustachian tube function. METHOD: We acquired the tympanogram of 30 normal persons (60 cases) by means of classical tympanometry, Toynbee test and valsalva maneuvre respectively, the peak point of each tympanogram was P1, P2 and P3 corresponding P1 -P2 > 10 daPa or Pmax-Pmin > 15 daPa was accepted as normal. RESULT: 70% cases were accepted normal, whose peak point shifted obviously. CONCLUSION: The method is a simple, objective and practical one; If the peak point shifts obviously, the function of ET is normal; On the contrary, if the function of ET may be abnormal, other tests should be performed. PMID- 12541490 TI - [The detection of Epstein-Barr virus in middle ear effusion of secretory otitis media]. AB - OBJECTIVE: To study the involvement of Epstein-Barr Virus (EBV) in the pathogenesis of secretory otitis media (SOM). METHOD: Epstein-Barr Virus (EBV) in blood serum, oral gargle and middle ear effusion was detected by polymerase chain reaction (PCR) method in 34 patients with secretory otitis media and 20 normal subjects. RESULT: The detection rate of EBV in serum and oral gargle in the SOM group was remarkably higher than that in normal control group (P < 0.01). EBV positive rate of middle ear effusion was higher than that of serum group in SOM individuals. CONCLUSION: EBV plays a role in the pathogenesis of secretory otitis media (SOM). PMID- 12541491 TI - [Benign paroxysmal positional vertigo following radiotherapy for nasopharyngeal carcinoma (report of 3 cases)]. AB - OBJECTIVE: To investigate the effect of ionizing radiation on the vestibule of the inner ear. METHOD: Three patients with benign paroxysmal positional vertigo (BPPV) accompanied by hearing loss following radiotherapy for nasopharyngeal carcinoma (NPC) were reported. Particle repositioning maneuver (PRM) was used as treatment for BPPV, and salvia miltiorrhiza tablets were taken orally to treat hearing loss. RESULT: Two of three cases with BPPV recovered and one was effective; Mean phone frequency hearing increased 10 dB compared with that before therapy. CONCLUSION: Occurrence of BPPV in this series may be related to radiation damage on the vestibule; PRM is a effective modality for the patients with BPPV. PMID- 12541492 TI - [Influence of chronic sinusitis on middle ear function]. AB - OBJECTIVE: To study the effect of range size and duration of chronic sinusitis on middle ear function. METHOD: Eustachian tube function, tympanogram and air hearing threshold were measured with an acoustic impedance audiometer and pure tone audiometer in 56 patients (112 ears). RESULT: Eustachian tube function was disturbed in 48.3% cases, tympanogram was abnormal in 42.0% cases, and air hearing threshold was increased in 33.9% cases. Comparison of middle ear function between anterior sinusitis and pan-sinusitis showed significant difference (P < 0.01), middle ear function between the course of less 5 years and of more than 5 years had significant difference (P < 0.05). CONCLUSION: The damage of middle ear function become more serious with the extent of lesion and with the longer course of disease in paranasal sinuses. PMID- 12541493 TI - [Extramedullary plasmacytoma of the head and neck: a clinicopathologic, immunohistochemistry study]. AB - OBJECTIVE: To investigate the relation between histopathological feature, clinical behavior characters, immunoglobulin (Ig) secretory function and prognosis of the extramedullary plasmacytoma (EMP) of the head and neck. METHOD: 12 cases of EMP in the head and neck region were studied retrospectively. The clinicopathological records from January, 1977 to December, 1996 were reviewed. Immunohistochemical (IHC) stains for detecting intracytoplasmic immunoglobulin kappa (IgG kappa) and lambda (IgG lambda) were carried out. RESULT: 10 cases of the patients were male and 2 female; Their ages ranged from 32 to 67 years with an average of 57 years. The tumors were located in the nasosinus (6), nasopharynx (2), Larynx (3) and ear (1). Low-(I), intermediate (II) and high-grade (III) tumors were 3, 3 and 6 cases respectively. Two patients with EMP of nasopharynx were treated with radiotherapy, in others radiotherapy was administered after complete surgical resection. Follow-up time ranged from 10 to 76 months. IHC showed monoclonality of the plasma cell and was positive for IgG lambda (6 cases) and IgG kappa (6 cases, conferming the identical clonal origin of the plasma cells. The 5-year survival for grade I, II and III were 75.0%, 62.0% and 37.6%, 37.5% and 68.50% for IgG lambda positive and IgG kappa positive cases, respectively. There are statistically significance between them (P < 0.01). CONCLUSION: EMP in head and neck region are rare plasma cell tumors. IHC play an important role in differential diagnosis. The combination of surgery and radiotherapy proved to be an adequate and effective treatment. The grading and IgG secretory function may be valuable for predicting the prognosis of EMP. PMID- 12541494 TI - [Thyroglossal duct cyst(fistula) operation and recurrence prophylasis]. AB - OBJECTIVE: To explore what leads to recurrent thyroglossal duct cyst (TGDC) following operation and how to improve the operation efficiency. METHOD: 32 patients with TGDC (including 8 recurrent cases) were performed operation for the second time. The cases who had infective signs received anti-inflammatory treatment prior to operation. Judge the relationship between thyroglossal duct cyst and hyoid bone before managing hyoid bone. The methylene blue was injected into TGDC for following up. Resect the small branches according to Sistruk procedure under the operation microscope. The residue underwent marsupiatization, electrocoagulation or radio frequency according to finger palpation. RESULT: 32 cases were successful with the operation and did not show any signs of recurrence after one-year following up. CONCLUSION: The recurrence of TDGC may be perfectly prevented as long as comprehensive treatment is carefully performed during the operation. PMID- 12541495 TI - [HSP70 expression of middle ear mucosa in acute suppurative otitis media]. AB - OBJECTIVE: To study HSP70 expression of middle ear mucosa in acute suppurative otitis media. METHOD: An animal model of acute middle ear infection was established by inoculating Klebsiella Pneumoniae into middle ear of guinea pigs. Animals were sacrificed 1, 3, 5 and 7 days after inoculation. The expression of Hsp70 epitope related proteins in middle ear mucosa were analysed by modified western blot test. RESULT: Very light 70 kD bands were recognized by anti Hsp70 monoclonal antibody in the unstressed group. However, middle ear mucosa of the stressed group not only showed relatively strong 70 kD bands, but also expressed more intensely 31 kD and 17 kD bands. The infected mucosa expressed one 70 kD band at 1st day, the strongest band at 3rd day, falling at 5th day; 31 kD was recognized two bands at 3rd day, the strongest bands at 5th day, falling at 7th day; two bands of 17 kD were detected in all days, with the strongest bands at 7th day. CONCLUSION: The results suggested that middle ear mucosa expressed Hsp70 and Hsp70 epitope related proteins like 31 kD and 17 kD in acute otitis media. PMID- 12541496 TI - [The localization of glutamate in the chicken cochlea]. AB - OBJECTIVE: To prove that glutamate plays a role as transmitter in cochlear-herrus acusticus introduction. METHOD: Glutamate immunoreactivity (Glu-IR) was investigated in the cytoplasm of the hair cells ganglion cells, and nerve fibers in normal chickens by means of monoclonal antibody immunochemistry. RESULT: The tall hair cells reacted more strongly than the short hair cells. The supporting cells showed no positive stain. It was suggested that glutamate may be the transmitter or modulator in the chicken cochlea. CONCLUSION: Glutamate may act as transmitter in cochlear introduction. PMID- 12541497 TI - [Genetic epidemiologic study of mitochondrial DNA 7445A-->G mutation among non syndromic deafness in Chinese population]. AB - OBJECTIVE: To study the Mitochondrial DNA 7445A-->G mutation in nonsyndromic deafness patients in Chinese population. METHOD: Polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) were used to screen the mitochondrial DNA 7445A-->G mutation among 128 nonsyndromic deafness individuals from 32 pedigrees, 135 sporadic nonsyndromic deafness patients and 100 normal subjects. RESULT: The 7445A-->G mutation did not appear in the experiment. CONCLUSION: Incidence of the mitochondrial 7445A-->G mutation was lower than that of mtDNA 1555A-->G mutation in nonsyndromic deafness patients in China. PMID- 12541498 TI - [Application of the quantitation with curve-fitting method on chiral separation by high performance liquid chromatography]. AB - We propose for the first time that curve-fitting method should be applied to the quantitation of overlapping peaks in chiral separation by high performance liquid chromatography (HPLC). We have examined the separation of eight isomers of cyfluthrin on a chiral stationary phase (CSP), and found that in comparing with the perpendicular-drop method, the accuracy of quantitation of overlapping peaks with curve-fitting was much higher. Generally, errors in peak area measurement using the perpendicular-drop method become larger as the degree of peak overlap increases. Meanwhile, by using curve-fitting method errors have no apparent difference for different degrees of peak overlap. Generally, for the same chiral compounds, although the resolutions are different with different mobile phase strength, the results of peak areas are the same by using curve-fitting method. PMID- 12541499 TI - [Direct optical resolution of acidic biphenyl drugs by high performance liquid chromatography on tris(3,5-dimethylphenylcarbamate) of cellulose]. AB - A chiral stationary phase was prepared by coating cellulose-tris(3, 5 dimethylphenylcarbamate) onto aminopropylated silica gel. A series of enantiomeric acidic biphenyl drugs were directly resolved on the chiral stationary phase (CSP) by normal-phase high performance liquid chromatography (HPLC). A hexane-2-propanol eluting system containing 1% of trifluoroacetic acid was used as mobile phase. Efficient optical resolution of the acidic biphenyl drugs has been attained. The factors that influence chiral discrimination such as structural characeristic of the samples and mobile phase were investigated. An interaction model between the stationary phase and the samples was discussed. The results showed that efficient optical resolution of racemic carboxylic acids could be attained by normal-phase HPLC on CSP using a hexane-2-propanol eluting system containing 1% of trifluoroacetic acid. PMID- 12541500 TI - [Screening for limonoid glucosides in Citrus grandis L. Osbeck by high performance liquid chromatography-electrospray ionization/mass spectrometry]. AB - A screening method for limonoid glucosides in the peel of Citrus grandis L. Osbeck, utilizing high performance liquid chromatography (HPLC) with diode-array detection (DAD), interfaced to electrospray ionization-mass spectrometry (ESI MS), has been developed. The methanol extract of citrus peel was preseparated with conventional polymer adsorption column and weak base anion ion-exchange column and then the eluate from the column was analysed by LC-ESI/MS. In this paper, the UV-Vis spectra and the mass spectra provide structural information without the necessity of isolating the individual compounds. Two major limonoid glucosides-obacunone glucoside (OG) and nomilin glucoside (NG) were identified in the methanol extract of the peel of Citrus grandis L. Osbeck. Then the two limonoid glucosides were taken as the target and isolated by means of preparative HPLC on a C18 reversed-phase column with an acidic acetonitrile-water mobile phase. The structures of OG and NG were further confirmed by NMR spectrometry. The results show that this method is rapid, convenient, sensitive and very useful for screening bioactive natural products. PMID- 12541501 TI - [Capillary gas chromatographic analysis and mass spectrometric identification of modified DNA bases 5-methylcytosine and 8-hydroxyguanine]. AB - The purpose of this paper was to establish a method for the determination of modified bases, 5-methylcytosine and 8-hydroxyguanine, in DNA by GC/FID. The experimental conditions were explored systematically for the quantitative analysis of these two modified bases, and the components were identified by GC/MS. The results showed that the variant components in DNA treated with Fenton's reaction can be derivatized and separated successfully. The relative molar reactive factors of 5-methylcytosine and 8-hydroxyguanine were 3.0 and 1.3, respectively. The sensitivity for them were 5.50 x 10(9) mV.s/g and 7.59 x 10(10) mV.s/g, respectively, while their detectable limits were 36.4 pg/s and 15.8 pg/s, respectively. The coefficients of variation for gas chromatograph were less than 5%, for derivatization, less than 6%, and for the whole analysis process, less than 20%. PMID- 12541502 TI - [Study on the separation of trace oxoanions by ion chromatography]. AB - It was the first time that bifunctional (cation exchange/anion exchange) stationary phase was used for the analysis of trace oxoanions by ion chromatography. The influence of sodium phosphate concentration and eluent pH on their separations was studied. An eluent of 7.5 mmol/L Na2HPO4(pH 9.3) was found to be the most suitable for the analysis of SeO3(2-), HAsO4(2-), SeO4(2-), WO4(2 ), MoO4(2-), GeO3(2-) and CrO4(2-) simultaneously. An eluent of 0.75 mmol/L Na2HPO4(pH 9.3) was found to be the most suitable for the analysis of IO3-, H2AsO3-, BrO3-, NO2- and NO3- simultaneously. The detection wavelength was 204 nm. Four other anion separation columns (AS4A-SC, AS9-SC, AS11 and PAX 500), which had different structures and properties, were compared to CS5A column. The separation mechanism was also studied. PMID- 12541503 TI - Hydroxy-dibenzo crown ether stationary phase for capillary gas chromatography using sol-gel technology. AB - A new kind of crown ether, OH-dibenzo-14-crown-4 (OH-DB14C4), is prepared and coated onto the fused silica capillary by sol-gel process. Chromatographic characteristics including column efficiency (> 3,000 plates/m), thermal stability (to 330 degrees C) and ability of deactivation are studied. The selectivity of new stationary phase is superior to sol-gel OH-terminal silicone oil (OH-TSO) for positional isomers of some aromatic compounds such as xylene, dichlorobenzene, nitrotoluene, nitrochlorobenzene. The new stationary phase has high sample capacity for separation of small molecular mass compounds: low-molecular-mass alcohols, ethers and ketones, short-chain fatty acids and volatile amines. PMID- 12541504 TI - [Recent advances of silica packing in high performance liquid chromatography]. AB - Developments of silica packings in high performance liquid chromatography are reviewed with 65 references. Preparative and modification methods of silica packings are introduced. The physical characterization on silica surface is elucidated. The applications of silica packings in HPLC are outlined and the trends of silica packings in China are described. PMID- 12541505 TI - [Development of integrated capillary electrophoresis chips]. AB - Capillary electrophoresis in its different operation modes has been demonstrated that it is suitable to be integrated into a planar microdevice which is called integrated capillary electrophoresis chip (ICE chip) or microchip. The use of microfabrication and micromachining allows the integration of complex channel nets and functional units such as pre- and post-column reaction chambers on a single chip, providing good sensitivity and high separation efficiency with low reagent consumption and short analysis time. In the last 10 years an important number of publications have been reported in this area. In the present paper, the development and applications are systematically reviewed with 39 references. PMID- 12541506 TI - [Prediction of output fraction from single layer to commingling oil well by high performance liquid chromatography(HPLC)]. AB - For crude oil from the same source, there is no obvious difference in chromatographic fingerprint of total hydrocarbons. But an evident difference in the concentrations of the series compounds of naphthalene and phenanthrene in oils has been found. Based on the fact, a new method for determine production allocation in commingled production, has been proposed. In this method, the series compounds of naphthalene and phenanthrene are selected as geochemistry indicator and are pretreated with aluminum oxide-silica gel column, and their relative concentration is determined by HPLC. Being verified by artificially mixing samples, the method can be used for the oils that can not be differentiated with the chromatographic fingerprint of total hydrocarbons, and error between predicted value and real value is 8% and its accuracy is better than that of the gas chromatographic fingerprint method. Based on the mixture principle, a model for prediction of output fraction of single oil layer is proposed. This model has been applied to production allocation calculation. PMID- 12541507 TI - [Determination of the adenosine triphosphate in myocardial tissue by ion-pair reversed-phase high performance liquid chromatography]. AB - ATP depletion is an important factor contributing to the incomplete recovery of ventricular function following ischemia and reperfusion. A simple and rapid method for determination of adenosine triphosphate(ATP) in myocardial tissue by RP-HPLC was developed. The chromatographic conditions were as follows: Spherisorb ODS2 column, V(40 mmol/L KH2PO4 and 5 mmol/L TBAP):V(methanol) = 74:26 (pH 6.2) mobile phase and UV detector at 259 nm. The ATP in myocardium was extracted with 0.4 mol/L HClO4. The calibration curve showed a good linearity in 5 mg/L-100 mg/L(r = 0.9998). The recovery ranged from 97.8% to 104.1%. The limit of detection was 2 mg/L. The verified results demonstrated that this method is precise, accurate and can be used for determination of ATP in myocardial tissue. PMID- 12541508 TI - [Preliminary study on the purification of IL-2]. AB - Interleukin-2 (IL-2) which is produced by T-lymphocytes is a protein molecule with the molecular weight of 15,420-Daltons. It has good functions in curing a lot of diseases such as cancer and AIDS even with a small quantity. A new method for purification of IL-2 by preparative high performance liquid chromatography is described. It was the first time to use hydroxyapatite as the packing to separate IL-2. IL-2 has one disulphide bridge (between 58th Cys and 105th Cys) and one free cysteine. In order to prevent aggregation and formation of wrong disulphide bond, 50 mumol/L Cu2+ and 1.5 mol/L guanidine hydrochloride was used for autoxidation and denaturing. Then we used mainly two chromatographic steps: First, Sephadex G-25 Gel permeation chromatography was used to separate guanidine hydrochloride and small molecules. Second, large scale preparative HPLC was used to separate impure protein from IL-2. Gradient elution was performed with phosphate buffer (pH 6.8). Its purity was examined by SDS-PAGE. Its activity reached 1 x 10(6) U/mg by CTLL-2 cell MTT method. The results showed that it is a suitable method for the large-scale purification of IL-2. PMID- 12541509 TI - [Research of indole-3-acetic acid biosynthetic pathway of Klebsiella oxytoca SG 11 by HPLC and GC-MS]. AB - The plant growth promoting bacteria are closely associated to plant. The bacteria are used to adhering to plant rhizoplane, promoting plant growth by fixing nitrogen from atmosphere, secreting stimulating substances or producing antagonistic to plant pathogens. It was indicated that the biological nitrogen fixation played an important role in plant growth promoting function. In fact, it was verified recently by overall research that IAA does it. Therefore research of IAA production and biosynthetic pathway of plant growth-promoting bacteria is much more important. The various ways of IAA production indicated the strong or weak promoting function of bacterium to plants in general. The purpose of this paper is to determine whether IAA exists in cultured medium of Klebsiella axytoca SG-11 and biosynthetic pathway of IAA, in order to opimize cultural conditions for IAA production. Klebsiella axytoca SG-11 is a plant growth promoting bacterium, isolated from rice rhizoplane, which can fix nitrogen. The supernatant of SG-11 cultured medium determined by HPLC showed that 47.4 mg/L of IAA existed in LB medium and 1.2 mg/L of IAA, in basal medium. IAA in metabolite was identified by GC/MS as well. The intermediate determination of tryptamine, indole 3-acetamide, tryptophol and indole-3-acetonitrile indirectly indicated that IAA was biosynthesized in a pathway of indole-3-pyruvic acid. Meanwhile, tryptophol in metabolite of SG-11 was verified by GC/MS. The direct intermediates of indole 3-pyruvic acid and indole-3-acetaldehyde in the pathway can not be determined, because both are unstable under normal condition. As reversible conversion existed between indole-3 pyruvic aldehyde and tryptophol, the presence of tryptophol also proved the pathway of indole-3-pyruvic acid in the synthesis of IAA by Klebsiella axytoca SG-11. The results laid basis for further research of plant growth-promoting function of the bacterium. PMID- 12541510 TI - [Analysis of carbon number and ethylene oxide number distribution in alcohol ethoxylates by HPLC/electrospray ionization mass spectrometry]. AB - Alcohol ethoxylates (AEO) of the formula RO(CH2CH2O)nH are an increasingly important class of nonionic surfactants. They are not single component but a complex mixture with a distribution of alkyl and ethoxylate groups. The average number of ethoxylate groups can greatly influence the characteristics of surfactants. High performance liquid chromatography (HPLC) was applied to separate alcohol ethoxylates(AEO) by alkyl and ethoxylate groups. Coupled with on line electrospray mass spectrometry, chromatographic peaks were assigned according to their mass spectra. Using this method, we succeeded in determining the distribution of LION AEO3 (Dobanox25c), which has an alkyl chain with 12, 13, 14 and 15 carbon atoms. Its ethylene oxide numbers vary from 1 to 12. The reason of distortion of the pseudo-molecule-ion intensity is also explained. The response of the MS detector for AEO species increased exponentially as the EO number increased from 1 to 6, after that, the increasing trend became moderate. PMID- 12541511 TI - [Determination of elemenyl piperidine salt in water by capillary gas chromatography]. AB - beta-Elemene and its derivatives were hopeful new antitumor drugs. A method for determination of elemenyl piperidine salt in water has been developed with capillary gas chromatography. The samples were neutralized by sodium carbonate and extracted by n-heptane. n-Octadecane was used as the internal standard. The linear range was from 10 g/L to 100 g/L, with a correlation coefficient of 0.9998. The average recovery of the method was 99.96% and the RSD was 0.46% (n = 5). The coefficient of variation was less than 2%. The method is simple, rapid, accurate and can be applied to the determination of the nitrogen-containing compounds of beta-elemene. PMID- 12541512 TI - [Determination of fatty acids in shark cartilage by GC-MS using microwave assisted digestion and derivatization]. AB - A rapid microwave-assisted digestion and derivatization method for the determination of fatty acids in shark cartilage by GC-MS was developed. The optimum conditions for digestion and derivatization were studied in detail using orthogonal design. The digestion and derivatization were accomplished in 4 minutes at 600 W microwave power using HCl-methanol (1:4, V/V) as digestion and derivatization solvent, and the extraction of the target analytes could be carried out simultaneously. This method is rapid, solvent-saving, and particularly suitable for the rapid determination of fatty acids in solid samples. PMID- 12541513 TI - [Stability of fatty acids in stored oyster]. AB - The contents of twenty-three kinds of fatty acids in the lyophilized oyster that stored for 0, 15, 30, 45, 60, 75, 90 days were studied. Oyster fat was extracted from the powder by means of SFE. The extraction was performed for 40 min at a pressure of 37 MPa and a temperature of 50 degrees C with supercritical carbon dioxide containing 8% (V/V) ethanol at a flow-rate of 2 mL/min as liquid carbon dioxide, and the recovery of oyster fat by extraction was over 99%. After being extracted, fat was esterified with potassium hydroxide and methanol. Methyl esters of fatty acid were separated and determined by 30 m x 0.25 mm i.d. x 0.25 micron HP-INNOWax capillary column and MS detector. The injector temperature was 220 degrees C. The column temperature was programmed from 150 degrees C (1 min) to 200 degrees C at 10 degrees C/min and then from 200 degrees C to 250 degrees C at 2 degrees C/min. Twenty-three peaks were identified with gas chromatography/mass spectrometry, and quantified with area normalization method. The variations of contents of them were shown. During storage, the contents of saturated fatty acids and mono-unsaturated fatty acids were getting higher, and those of polyunsaturated fatty acids were getting lower. The decrease of them was gradual, and there was no special period of stability. And the stability of fatty acids in oyster related to the degree of unsaturation of them. The higher the unsaturation the lower the stable it was. After being stored for 90 days, the content of EPA decreased from 16.94% to 5.43% and that of DHA from 9.25% to 2.86%. PMID- 12541514 TI - [Preparation of gas chromatographic capillary columns with beta-cyclodextrin polymer stationary phase modified with methyl phenyl silicone(OV-17)]. AB - An insoluble beta-cyclodextrin polymer cross-linked with epichlorohydrin was prepared. beta-Cyclodextrin polymer (0.26 g) was treated for 0.5 h by a ultrasonator in the mixed solvent (10 mL) of isopropyl ether, methylene dichloride and benzene with a volume ratio 2:1:2, and a 26 g/L colloidal stationary liquid was prepared. It was coated on a fused silica capillary by superdynamic coating method. The capillary column was of high column efficiency of above 3,000/m with good thermal stability and moderate polarity. Enantiomers of xylene, dichlorobenzene, alpha-pinene, beta-pinene, ethyl lactate, alpha phenyl ethanol, 1, 2-propanediol, alpha-lonone, methyl alpha-chloropropionate and gamma-valerolactone were separated on the capillary column. The experimental results showed that beta-cyclodextrin polymer treated by a ultrasonator is a capillary chromatographic stationary phase with good performance and high selectivity. PMID- 12541515 TI - [Determination of residues of 20 kinds of organochlorinated pesticides in oils, fruits and vegetables by wide-bore capillary gas chromatographic column]. AB - A rapid and high performance gas chromatographic method for the determination of 20 organchlorinated pesticides in oils, fruits and vegetables has been developed. The samples of fruits and vegetables were extracted with petroleum ether-acetone, while those of oils were extracted with acetonitrile-hexane. The extract was purified by Florisil column with ethyl ether-hexane(15:85, V/V) or ethyl ether petroleum ether(15:85, V/V) as eluent. Chromatographic analysis was performed on wide-bore capillary column with ECD detector. Satisfactory separation and detection of these organchlorinated pesticides are presented with the proposed method. The detection limits of this method were 1.0 ng/g-20.0 ng/g(S/N = 5), the recoveries 83.2%-106.8% and the relative standard deviations 2.0%-9.5%. PMID- 12541516 TI - [Purification of prothrombin in Nitschmann fraction III by membrane radial column ion-exchange liquid chromatography]. AB - A method is described for the purification of human prothrombin complex concentrate (PCC) from Nitschmann fraction III by membrane radial column ion exchange chromatography, which allows large sample volumes to be processed at low operation pressure. The Nitschmann fraction III (15 g) was mixed with 1,000 mL 0.06 mol/L Tris-HCl(pH 7.5). The centrifuged supernatant(10,000 r/min, 15 min, 20 degrees C) was applied onto a DEAE ion-exchange liquid chromatographic column(XK 16 DEAE fast flow Sepharose and DEAE membrane radical column chromatography) with almost the same excellent separation efficiency. The parameters of sample flow rate, elution flow rate and sample capacity were optimized. PMID- 12541517 TI - [Study on the characteristics and applications of a new hydrophobic interaction chromatographic packing using chitosan as support]. AB - Based on chitosan, by using valeraldehyd as ligand, a new hydrophobic interaction chromatographic (HIC) packing material was prepared by an improved method. The effects of salt concentration, temperature, pH on the adsorption of bovine serum albumin(BSA) were investigated. The results showed that the adsorption behavior of protein by packing material is in agree with the theory of hydrophobic interaction. The alpha-amylase could be separated by HIC with high recovery of enzymatic activity. PMID- 12541518 TI - [The separation of three basic drugs with carboxymethyl-poly-beta-cyclodextrin as chiral additive by capillary electrophoresis]. AB - Five cyclodextrin derivatives, namely phosphate ester beta-CD, carboxymethyl-beta CD(CM-beta-CD), bis-[6-oxygen-(beta-carboxymethyl-succinic acid-4-ester)] beta CD(F-CM-beta-CD), beta-CD polymer(P-beta-CD), carboxymethyl-poly-beta-CD (CD-P beta-CD), were used as chiral selector for separation of three basic drugs, lobeline, thiopendonesodium, flunarizine by capillary zone electrophoresis (CZE). All the five cyclodextrins have chiral separation ability to lobeline. P-beta-CD and CM-P-beta-CD have chiral recognition to thiopentonesodium and flunarizine. The results indicate that via optimizing separation conditions by varying pH and the concentration of chiral selectors the three racemic drugs could be baseline separated with 2% P-beta-CD or 0.5% CM-P-beta-CD in the buffer of 30 mmol/L Tris H3PO4. And the best resolution ranging from 4 to 35 with CM-P-beta-CD as chiral additive was obtained within 10 min. The results were much better than those reported in other references. PMID- 12541519 TI - [Preparation of immobilized metal-chelated affinity membrane and its application to purification of Cu/Zn-superoxide dismutase]. AB - Immobilized metal-chelated affinity membranes were prepared using macropore cellulose filter paper as matrix. The matrix was treated with alkaline, activated with epichlorohydrin and coupled with iminodiacetate sodium, and then Cu2+ was immobilized. Cu/Zn-superoxide dismutase(SOD) was purified with prepared affinity membranes. The special activity of the purified Cu/Zn-SOD was increased 10.7 times from 645 U/mL to 6,882 U/mL. The protein recovery and the activity recovery were 92.3% and 985%, respectively. A new method to prevent the leaking of metal ion was developed and Cu2+ concentration in the effluent was lowered to 86 micrograms/L. PMID- 12541520 TI - [Identification of synthetic adhesives by simultaneous pyrolysis methylation gas chromatography]. AB - At present, pyrolysis gas chromatography (Py-GC) is an effective analytical method for trace synthetic adhesives. Synthetic adhesives are usually composed of polar components. Thus, the polar pyrolysis products are normally difficult to measure by Py-GC due to their partial or complete adsorption in the pyrolysis zone, injection system or capillary column and the accuracy of results is seriously affected. Simultaneous pyrolysis methylation gas chromatography (Py-Me GC) is a fast analytical method which was developed in recent years for polar compounds. The five synthetic adhesives have been identified by using Py-Me-GC in our study. Analytical conditions were: filament pyrolysis apparatus, flame ionization detector (FID), gas chromatograph, FFAP capillary column (30 m x 0.25 mm i.d.) and a quaternary ammonium salt as methylating agent. Major peaks of the products of pyrolysis were analysed qualitatively by GC/MS and comparisons were also made between the results by Py-Me-GC and those by Py-GC. The information gained from the components of substances by Py-Me-GC is more than that by Py-GC. The method is suitable for identifying synthetic adhesives of trace evidence in forensic science. PMID- 12541521 TI - [Determination of three active ingredients in food additive sucrose fatty acid ester by TLC scanning]. AB - A method of separation and quantitation of monoester, diester and triester in food additive sucrose fatty acid esters by TLC with dual-wavelength TLC scanner in the sawtooth scanning mode is described. The detective wavelength was 530 nm and the reference wavelength was 700 nm. A mixture of chloroform, methanol, acetic acid and water (80:10:8:1, V/V) was used as mobile phase. The calibration curve was linear over a range from 4 micrograms to 60 micrograms with correlation coefficients of 0.9949-0.9980. The average recoveries were 96.45%-98.73% (n = 3, RSD = 2.7%-3.2%). This method is accurate, simple and dependable, with a wide range of linearity. It has been applied to the analysis of various samples and can be used for the quality control of the food additive sucrose fatty acid ester. PMID- 12541522 TI - [Analysis for monobromo- and dibromo-pentaerythritol by gas chromatography]. AB - A method for separation and determination of dibromo-pentaerythritol (DBPT), monobromo-pentaerythritol (MBPT), and tribromo-pentaerythritol (TBPT) by gas chromatography was developed. The conditions for determination were: BPX-70 capillary column, operated at 240 degrees C, with both FID detector and injector at 280 degrees C. Xylitol was the internal standard. Linear regression equations were YDBPT = 0.0423X-0.0124 (r = 0.9998) and YMBPT = 0.0634X-0.0835(r = 0.9966). This method is rapid, simple and accurate. PMID- 12541524 TI - [Process monitoring the synthesis of N-dichloroacetyloxazolidine by thin-layer chromatography]. AB - A thin-layer chromatographic procedure has been developed for the monitoring and identification of the synthesis of N-dichloroacetyloxazolidine by dichloroacetyl chloride, an alcoholamine and a ketone. The determination of reaction was made on the plates at room temperature. The reaction liquid was spotted on the silica gel G thin-layer plate and developed using solvent system, dichloromethane methanol(10:1, V/V), until the solvent front migrated 10 cm from the starting spot. Then, the separated spots(raw materials and product) were scanned in iodine. About 1 hour, the raw material spots disappeared gradually and the product spots became clear, then the reaction had basically completed. Compounds A, B, C and D had Rf values of 0.71, 0.76, 0.75 and 0.79 respectively. The repeatability of Rf value was good. The TLC procedure has also been developed for the identification of the recrystallization of crude product. After recrystallization, there was one spot on the plate. Then, the products were confirmed by IR, 1HNMR and elemental analysis. The separation conditions were easily obtained. This method is simple, rapid, sensitive and reproducible. PMID- 12541523 TI - [Determination of alpha-naphthylacetic acid by high performance liquid chromatography]. AB - A high performance liquid chromatographic method for the determination of alpha naphthylacetic acid has been established on a Hypersil C18 column(4.6 mm i.d. x 250 mm, 5 microns), with the mobile phase of methanol-water-phosphoric acid(60:40:0.35, V/V), and UV detection at 272 nm. The linear correlation was observed within the range of 16 mg/L-1,000 mg/L(r = 0.9997). The RSD was 0.8%(n = 5). The method is simple and accurate. PMID- 12541525 TI - [Analysis and stability study on nimotop injections by RP-HPLC]. AB - A method was developed by using reversed-phase high performance liquid chromatography (HPLC) to analyze nimotop injections. Methyltestosterone was used as an internal standard. The separation was performed on a YWG C18 column with mobile phase of V (methanol):V (water) = 65:35 and detected at 238 nm. The linear concentration range of this method was 5.98 mumol/L-299.0 mumol/L. This method was simple, rapid and has been used to study the stability of nimotop injections. The experimental results showed that the nimotop was stable at higher temperature (50 degrees C) but unstable under light. Nimotop injection should be kept away from light. PMID- 12541526 TI - [Reuse of the print cartridge of chromatograph in data processor]. PMID- 12541527 TI - [Telomerase activity in human head and neck squamous cell carcinoma and adjacent tissues]. AB - OBJECTIVE: To investigate the telomerase activity in human head and neck squamous cell carcinomas and their adjacent tissues and to explore its possibility of being tumor biological marker. METHOD: Thirty-two patient samples of primary head and neck squamous cell carcinoma and 15 adjacent tissues were assayed using TRA PCR-ELISA for detection of telomerase activity. RESULT: Twenty-seven of 32 (84.4%) cancer samples and 5 of 15 (33.3%) adjacent samples were telomerase positive (P < 0.05, Chi-Square Test). The frequency of telomerase activation was lower in cancer without lymph node involvement (82.4%) than that with lymph node involvement(86.7%), but the difference has no statistical significance. Detection rate of telomerase activity in poor grading cancer (90%) is higher than that in moderate and high differentiation cancer (81.8%), although no statistical significance. No obvious correlation was found between the telomerase activity and the other clinical parameters such as primary site and staging. CONCLUSION: The finding of telomerase activity in 84.4% of primary head and neck squamous cell carcinoma samples is consistent with the concept of telomerase playing a key role in tumorigenesis. Further study is needed to determine the usefulness of this enzyme as a new molecular marker. PMID- 12541528 TI - [Clinical study on treatment of early glottic carcinoma with different cordectomy]. AB - OBJECTIVE: To explore the advantage of KTP laser cordectomy in the treatment of early glottic carcinoma. METHOD: Follow-up observation on 30 patients who had KTP laser cordectomy and 18 patients who had laryneal split cordectomy was applied, then their postoperative laryngeal structure's change, the quality of voice and the quality of life (QOL) were compared and analyzed. RESULT: The patient group treated by KTP laser grew out a piece of new cord just at the place where it was excised after half a year, and the quality of voice was obviously improved near to norm, and they had higher QOL. On the contrary, the other group had no newly grown cord, their quality of voice was better in the first half year but was worse last out. Their QOL was lower than the laser group before decannulation and the same as the laser group after the decannulation. There are 2 cases recurrented in the laser group and 1 case recurrented in the laryneal split group, the two groups all had no metastasis and death in laryneal carcinoma by 2 4 years follow up postoperation. CONCLUSION: On the basis of effecting radical cure, the early glottic carcinoma treated with non-contact KTP laser with suspension laryngoscope can result in good recuperating in laryngeal structure, less injury and high QOL. So it makes micro-injury treatment in laryngeal carcinoma come true and better than the laryngeal split cordectomy. PMID- 12541529 TI - [Clinical study of effects of submaxillary transcutaneous electrical stimulation of genioglossus on obstructive sleep apnea syndrome]. AB - OBJECTIVE: To investigate the effect of submaxillary transcutaneous electric stimulation of genioglossus on obstructive sleep apnea syndrome (OSAS). METHOD: Nineteen patients (male) with OSAS were diagnosed by polysomnography previously. Fixed the provocative locus of genioglossus and its controlling nerve with local anatomy. Estimated obstructive level of pharyngeal cavity by observing muller's maneuver under fiberolaryngoscope. Determined the effects of electric stimulation on different levels of narrow pharyngeal cavity with control study method. RESULT: The effect of electrical stimulation was different in different narrow level. The best effect was found on the obstruction of retrotongue-base-pharynx level, data of breath disorder (P < 0.05), oxygen saturation (P < 0.05) and clinical symptoms were significantly improved. The data were partly improved on multilevel obstructions of retrotongue-base-pharynx or above, no improvement was found on the obstruction of retrotongue-base-pharynx level. Dormant index was not worse. CONCLUSION: Submaxillary electrical stimulation is effective to the treatment of OSAS and its curative effect perhaps has a close relationship with obstructive level of with upper-airway. Further research on the mechanism showed that stimulating upper-airway dilating muscle which formed mainly by genioglossus could push the tongue ahead and effectively open pharyngeal cavity. Maybe it is a reason that the best effect took place on the retrotongue-base-pharynx level. Sleep was not affected by electrical stimulation. PMID- 12541530 TI - [Surgical treatment for the recurrent patients after palatopharyngoplasty]. AB - OBJECTIVE: To study the surgical treatment for the recurrent patients after palatopharyngoplasty (PPP). METHOD: Five patients which were confirmed recurrence by sleep survey and examinations were reported. RESULT: The pharyngeal cavity was narrower obviously ten months after ppp than before ppp. The anteroposterior and transverse distances were (4.06 +/- 2.94) mm and (11.40 +/- 4.16) mm respectively. After dilatation, the distances were (11.90 +/- 3.25) mm and (21.00 +/- 4.18) mm separately, and were increased significantly than before (P < 0.01). Two patients were cured and three cases marked, and all patients were not recurrent post-operation with no complications. CONCLUSION: The recurrence was mainly concerned with the unsuitable manipulation in PPP. Selecting the cooperation and proper surgical treatment, the better effect was also obtained for the recurrent patients. PMID- 12541531 TI - [The study on the standardization of adults voice acoustic parameters by objective analysis]. AB - OBJECTIVE: To explore the characteristic of normal voice in different age-bracket of Chinese adults and to formulate the normal reference value suited to our country at voice assessment. METHOD: We divided the adults into the youth, the middle-aged and the old groups, and analyzed normal voice samples with Dr. Speech software and computer multimedia technique and compared the following 5 parameters (Jitter, Shimmer, NNE, Fo and SDFo) of different groups divided according to one's age, sex and vowel. RESULT: There is difference among the group of age, sex and vowel in most acoustic parameters in the adults. CONCLUSION: To initiate taking multi-vowels samples and using different normal reference value and multi-parameters analysis according to the difference of age bracket, sex and vowel in the adults. PMID- 12541532 TI - [Hemangioendothelioma of larynx]. AB - OBJECTIVE: To study the biological behaviour and treatment of larynx hemangioendothelioma. METHOD: One case of larynx hemangioendothelioma was reported and the relative literatures were reviewed. RESULT: The common symptoms of the disease include hoarseness and larynx obstruction caused by the enlargement of the tumor. The biological behaviour of the larynx hemangioendothelioma is low-grade malignant and recurred, between the benign hemangiomas and malignant angiosarcomas. CONCLUSION: The treatment of the hemangioendotheliomas is enlarged removal of the tumor by Majer-Piquet' operation. Chemotherapy and radiation are not valuable. PMID- 12541533 TI - [Change of Fo and vocal range of pathological voice]. AB - OBJECTIVE: To study the relationship between Fo and true or falsetto range in four phonation modes. METHOD: Testing Fo of lowest true voice, comfortable voice, highest true voice and falsetto voice. RESULT: In pathological voice, Fo of lowest true voice increases, Fo of highest falsetto voice reduces, true and falsetto voice range become thin and the change of falsetto voice range is more obvious. CONCLUSION: Fo of falsetto highest voice reduces, falsetto voice rang become thin in pathological voice. PMID- 12541534 TI - [Detection of HPV-DNA and humoral immunity in juvenile laryngeal papilloma]. AB - OBJECTIVE: To study HPV infection route and pathogenesis of juvenile laryngeal papilloma(JLP). METHOD: HPV-DNA of JLP was detected with PCR and PCR product dot blot hybridization. Serum Ig and complement 3(C3) was detected with nephelometry. RESULT: HPV total positive rate in JLP was 95%(19/20). HPV6 was 55%(11/20). HPV11 was 30%(6/20), and HPV6 + 11 was 10%(2/20). Serum IgG, IgA, IgM and C3 of JLP were normal, no significant difference between JLP and the control group(P > 0.05). CONCLUSION: HPV6 infection of JLP was in the majority. And the type was consistent with the type of female genital organ pointed condyloma. Humoral immunity was negligible in HPV infection. Pathologic picture of different HPV type infection was identical. PMID- 12541535 TI - [Comparison of four techniques in the treatment of little's area epistaxis]. AB - OBJECTIVE: To probe the most effective technique in the treatment of little's area epistaxis. METHOD: 851 patients were separately treated with four techniques. The effects of sclerosant mucosa injection, cryotherapy, CO2 laser and chemical cautery in the treatment of epistaxis were compared. RESULT: The effective rate of sclerosant mucosa injection is 92.4%, CO2 laser 75.8%, Cryotherapy 76.4%, Chemical cautery 73.8%. No side effect was found. CONCLUSION: Sclerosant mucosa injection is superior to the others. PMID- 12541536 TI - [Treatment of chronic hypertrophic rhinitis by Nd:YAG laser under nasal endoscope]. AB - OBJECTIVE: To evaluate the curative effect of the treatment of chronic hypertrophic rhinitis with Nd:YAG laser under nasal endoscope. METHOD: Fifty seven patients with chronic hypertrophic rhinitis were treated with Nd:YAG laser operation under nasal endoscope at our outpatient clinic. Some factors affecting operation effect were discussed. RESULT: In all patients, 10 cases (17.5%) were cured, 16 cases (28.1%) significantly improved, 28 cases (49.1%) improved, 3 cases (5.3%) unimproved and the total effective rate was 94.7%. No complication happened. CONCLUSION: Compared with the partial middle or inferior turbinectomy and YAG laser surgery without nasal endoscope. This technique has several advantages such as less damage, less hemorrhage, clear and clean operation field, easy to operate and so on. PMID- 12541537 TI - [Analysis of 19 cases adenoid cystic carcinoma of the nasal cavity]. AB - OBJECTIVE: To study the biological features of adenoid cystic carcinoma of the nasal cavity in order to provide a basis for its clinical treatment. METHOD: A retrospective analysis was made to 19 cases which were treated with operation combined with preoperative chemotherapy and radiation. RESULT: 5, 8, and 10 years' follow-ups conducted post treatment showed the local recurrence rates were 5.3%, 21.1%, and 31.6% respectively. CONCLUSION: Adenoid cystic carcinoma of the nasal cavity is highlighted because of high-degree local infiltration, the specific anatomy of the cavity, and the difficulty of wide-ranging surgical resection. Preoperative chemotherapy combined with radiation has the advantages of extinguishing the recessive cancerous foci, inhibiting the primary foci and lowering local recurrences and cancerous metastases. Thus it constitutes an effective way to deal with this malignancy. PMID- 12541538 TI - [Study on the association between HPV16/18 and expression of p53 protein in laryngeal papillomas]. AB - OBJECTIVE: To understand the relationship between the expression of p53 protein and HPV16/18 infection in laryngeal papillomas. METHOD: Using PCR and immunohistochemistry techniques examined the paraffin embedded tissues of 35 cases of laryngeal papillomas. RESULT: Showed that HPV16/18-DNA were found in 24 cases of laryngeal papillomas (68.8%); overexpression of p53 protein were detected in 19 cases (54.3%); both of HPV16/18-DNA and overexpression of p53 protein were demonstrated in 12 cases of laryngeal papillomas (34.3%). CONCLUSION: This study suggests that HPV16/18 infection and p53 gene mutation are associated with pathogenesis of laryngeal papillomas. The relation between HPV infection and p53 mutation in tissues of laryngeal papillomas remains to be clarified. PMID- 12541539 TI - [The location of inserting the electrode into the cochlear]. AB - OBJECTIVE: To determine the position of implanting electrode into the scala tympani. METHOD: The analogous operation of cochlear implantation were made in 50 temporal bones and the anatomic data were measured after the operation. RESULT: The position of implanting electrode is 2.77 mm below the vestibular window and 0.85 mm anterior to the round window. CONCLUSION: It suggests that this anatomic parameter provide a reference value to determine the position of cochlear implant electrode into the scala tympani. PMID- 12541540 TI - [The discrimination rate in the consonant by cochlear implanted]. AB - OBJECTIVE: To evaluate the effect of the discrimination rate in the consonant by cochlear implanted. METHOD: The four cases by cochlear implanted have rehabilitated with hearing speech for 1.0-1.5 years. RESULT: The discrimination rate in the four cases by cochlear implanted was 76.0% in the consonant, 94.1% in the vowel. CONCLUSION: The discrimination rate in common language of Chinese was lower than that of vowel. When the patients discriminated the consonants with the same formants feature, the chance of the auditory is relatively high. PMID- 12541541 TI - [Investigation report on the inherited characteristics of congenital preauricular fistula]. AB - OBJECTIVE: To further analyse the inherited characteristics of congenital preauricular fistula. METHOD: Analysed the familial incidence of 8 cases out of 73 (95 ears) which had been cured and investigated in our hospital in the past 16 years. RESULT: The disease could be inherited from either parents; It was of autosomal dominant in type; It was not sex-linked; They inherited either in the same side or both sides; It could be transferred between every other generations. CONCLUSION: All these were resulted in that congenital preauricular fistula was autosomal dominant inheritance with incomplete dominant. PMID- 12541542 TI - [The evaluation of eustachian tube function in patients suffered from SOM with C type curve]. AB - OBJECTIVE: To explore eustachian tube (ET) function in patients suffered from SOM with C type curve. METHOD: To test the tympanogram following toynbee test and valsalva manoeuvre respectively. The shift of peak point was quantified and the results were compared with normal subjects. RESULT: ET dysfunction was found in 52.38% of the patients, it was observed in 30% of the normal subjects (P < 0.05). CONCLUSION: ET dysfunction is one of the causes of SOM with C type curve, but not the only cause. PMID- 12541543 TI - [Reconstruction acusticus back wall and tympanoplasty with homograft costal cartilage]. AB - OBJECTIVE: To eliminate the back wall defect of external canal and opened mastoid cavity which was remained from radical mastoidectomy. METHOD: We performed the operation on 17 cases after radical mastoidectomy by using reconstruction of the canal wall and fill-in opened mastoid cavity and ossicular replacement prostheses made of homograft costal cartilage. The tympanoplasty had been made operated for all patients, 8 ears were PORP, the others were TORP. RESULT: All cases were followed-up for 6 months-2 years. The external canal of all patients are normal in appearance. The total effective rate of gain (> 15 dB) was 14 ears (82.4%). Only one patient whose tympanic membrane was perforated postoperation 3 months. CONCLUSION: Reconstruction of acusticus back wall and hearing after radical mastoidectomy with homograft costal cartilage is effective to prevent mastoid cavity infection and improve hearing. PMID- 12541544 TI - [The preoperative application of selective catheterization and embolism of the external carotid artery in treating tumors with enriched blood supply in the base of skull]. AB - OBJECTIVE: Embolizing the feeding vessels of the tumors with enriched blood supply in the base of skull in order to reduce the bleeding during operation. METHOD: Under digital subtraction angiographic control, superselectively catheterizing the external carotid artery, thus occluding the terminal branches of the feeding arteries of the tumor with particles of liquidized gelfoam. RESULT: Nine cases studied showed that the angiography of the feeding vessels of the tumor disappeared relatively. The bleeding decreased significantly during operation. The postoperative pathological findings showed that the entity of the tumor was congested with gelfoam. CONCLUSION: After figuring out the blood supply of the tumor, superselective catheterization of the external carotid artery followed by occlusion of the feeding vessels can reduce the size of the tumor and diminish the bleeding during operation, thus can improve the safety of the operation. It should be one of the crucial preoperation measures. PMID- 12541545 TI - [A contrast analysis of four phonation methods frequency between patients with glottis closure disturbance and normal human]. AB - OBJECTIVE: To find out frequency change parameters of four phonation methods (minimum, natural, maximum and falsetto maximum voices) and different maximum phonation persistent time (MPPT) of them in patients with glottis closure disturbance (GCD). METHOD: With frequency spectrum analysis, the frequencies of four phonation methods were studied in functional GCD patients (150 cases) and normal human (60 cases). RESULT: There were marked differences of voice frequencies between them not only in male but also in 5 female. And with the prolongation of MPPT, the voice frequencies also changed. CONCLUSION: The functions of both low and high voices decline in GCD patients. With the prolongation of MPPT, the functions of low and high voices strengthen in GCD patients. PMID- 12541546 TI - [Correlation analysis of laryngeal vocal efficiency and nasalance after vertical hemilaryngectomy]. AB - OBJECTIVE: To analyse the correlation between laryngeal vocal efficiency and nasalance for patients after vertical hemilaryngectomy. METHOD: The lately invented instruments, phono-laryngography and nasometer, which can objectively evaluate vocal function, were used to determine the laryngeal vocal efficiency and nasalance in 30 patients with laryngeal carcinoma treated primarily by vertical hemilaryngectomy. RESULT: The results showed that laryngeal vocal efficiency and nasalance have significant positive correlations in all subjects (r = 0.47). CONCLUSION: Change of laryngeal vocal function influence the value of nasalance. PMID- 12541547 TI - [Clinical analysis of 35 patients with orbital maxillary zygoma complex fractures]. AB - OBJECTIVE: To assess the treatment result of reposition and fixation to orbital maxillary zygoma complex fractures. METHOD: The records of 35 patients, who were treated in 1996-1998 at our hospital, were analyzed retrospectively. The patients of open fracture operated debridement, at the same time, operated reposition and fixation. The patients of closed fracture operated hairline approach and sublabial approach or use two approach for reposition and fixation. RESULT: We had 35 patients of operation. After operation, two of them had malocclusion, one of them had facial deformity, two of them had restriction of mouth opening. The treatment results of the rest appeared to be reasonably satisfactory. CONCLUSION: To know topographic anatomy, earlier reposition and sturdy fixation is the key for curing orbital maxillary zygoma complex fractures. PMID- 12541548 TI - [Report of 4 cases for treating optic nerve inflammation through total ethmoidectomy under nasal endoscope]. AB - OBJECTIVE: The curative effect of treating optic nerve inflammation through total ethmoidectomy under nasal endoscope is evaluated. METHOD: The operations through total ethmoidectomy under nasal endoscope for 4 patients who had optic nerve inflammation has been done. RESULT: Two patients have apparent effect while the other two patients are effective, and the symptoms such as snuffle, eye distension and headache are well remitted. CONCLUSION: Towards the sufferer who had both optic nerve inflammation and intercurrent nasosinusitis, when conservative treatment was invalid or the patient was relapsed after surgery, it is necessary to do the operation in season and its effective rate is satisfactory. Total ethmoidectomy under nasal endoscope is safety and reliable. PMID- 12541549 TI - [Clinical analysis of 20 cases of adenoidal hypertrophy in adult]. AB - OBJECTIVE: To discuss the diagnosis and properly treatment to the adult adenoidal hypertrophy. METHOD: To do nasal endoscopic and pathologic exam to all the 20 symptomatic patients before conservative or operative treatment. RESULT: The results of exam under endoscope were that 8 patients were hypertrophy I, 8 hypertrophy II, 4 hypertrophy III. Pathologic exam: 7 acute and chronic inflammation, 13 chronic inflammation with hyperplasia. TREATMENT: 10 of them received conservative treatment all have negative result. 6 received traditional operation, 2 of them reoccurred within 1 year. 4 received adenoidectomy and microwave treatment under nasal endoscope, all have good result in the next 1 year. CONCLUSION: Some adults have different sizes of adenoidal hypertrophy. This hypertrophy is different with children's under the pathologic microscope. Operative treatment especially the nasal endoscopic adenoidectomy and microwave treatment is a safe and effective method. PMID- 12541550 TI - [The proliferating cells in the rat endolymphatic sac]. AB - OBJECTIVE: To study the cellular proliferating activity of the endolymphatic sac. METHOD: 10 healthy adult S-D rats were used for the experiment. After administration of 5-bromo-2'-deoxyuridine (BrdUrd) intraperitoneally, the rats were sacrificed and the temporal bones were removed, then the decalcified samples examined immunohistochemically by using monoclonal antibodies against BrdUrd. RESULT: Scattered BrdUrd+ cells were detected within the endolymrhatic sac (ES) whereas labelled cells were not fonnd in the cochlea. CONCLUSION: Possessing a low rat of mitotic activity, the ES may be an important the sole site for the cell-renewing in the inner ear. PMID- 12541551 TI - [Distribution of cholinergic efferent vestibular neurons]. AB - OBJECTIVE: To investigate the effect and distribution of cholinergic efferent vestibular neurons in normal wistar rat. METHOD: HPR retrograde tracer and Immunocytochemical techniques were used. RESULT: Results show that AchT immunoreactive neurons locate in dorsolateral and dorsomedial to genu of the facial. Few neurons scatter in the parvocellular nucleus. CONCLUSION: As a neural transmitter, acetylcholine distribute in efferent vestibular neurons and play an important role. PMID- 12541552 TI - [Thermodynamic characteristics of stoichiometric displacement linear parameter log I in reversed-phase liquid chromatography]. AB - Based on the physical meaning of each term in the linear parameter log I (a constant correlating the affinity of solute to stationary phase) of stoichiometric displacement model for retention(SDM-R) of solute, the thermodynamic characteristics of log I of solute in reversed-phase liquid chromatography (RPLC) were investigated theoretically. This point was tested by experimental data with two linear relationships: (1) plot of log I vs 1/T (T is absolute temperature) with apolar, polar small solutes and proteins; (2) plot of log I vs log Po/w (partition coefficient of solute between n-octanol and water) with apolar and polar small solutes. From the comparison of the magnitude of partition coefficient between small solute and proteins, a phenomenon that the resolution of small solutes depends on column length but that of proteins almost to be independent of column length is quantitatively explained. PMID- 12541553 TI - [Retention behavior of some polycyclic aromatic hydrocarbons on alkylphosphonate modified magnesia-zirconia composite stationary phase for reversed-phase liquid chromatography]. AB - An alkylphosphonate-modified magnesia-zirconia composite stationary phase (PZMS) for reversed-phase liquid chromatography was prepared. The reversed-phase chromatographic performance of the new packing material was investigated with some polycyclic aromatic hydrocarbons (PAHs) as solutes. The effect of the methanol content in the mobile phase on the retention of PAHs was examined. The logarithmic capacity factor of PAHs was decreased linearly with increasing volume percent of the methanol in the mobile phase, which is similar to that observed on Zorbax ODS. The capacity factor of PAHs on the new stationary phase was also correlated to their partition coefficient between n-octanol and water (log P). A good linear relationship between them was obtained. These results show that the new material can behave as a reversed-phase packing. The separations of phenanthrene and anthracene, and terphenyl isomers were investigated on the new stationary phase and the Zorbax ODS, respectively. The retention of the solutes on the Zorbax ODS is greater than that on the new stationary phase, but the separation selectivity of the solutes on the new stationary phase is better than that on the Zorbax ODS. The better separation selectivity on the new stationary phase may be contributed from the Lewis acid-base interaction between the solutes and the residual Lewis acid sites on the stationary phase. PMID- 12541554 TI - [Separation and determination of tyramine by reversed-phase high performance liquid chromatography]. AB - The method for separation and determination of tyramine by reversed-phase high performance liquid chromatography has been established. The effects of mobile phase on mechanism of separation based on the ion interaction are discussed. Using C8 alkyl bonded phase and 40% methanol-water solution containing Trisperchlorate as pH buffer (adjusted pH to 7.9) and ion interaction reagent and applying p-toluene sulfonamide as internal standard substance, the tyramine obtained from decarboxylation of p-tyrosine was determined. The RSD of determination was 0.66%(n = 11). The recoveries of sample were between 99.33% and 100.38%. PMID- 12541555 TI - [Study of the crystalline behaviors of polymer by inverse gas chromatography]. AB - An investigation was made on crystallinity(Xc) and melting temperature(Tm) of the crystalline polymer polyethylene glycol(PEG) by inverse gas chromatography(IGC). The influence of the property of the probe and the coating weight of PEG in stationary phase on the data determined is discussed. And IGC data are compared with those got by differential scanning calorimetric (DSC) experiment. Results showed that IGC is a very useful technique for measuring the melting point and the degree of crystallinity of polymer samples since it does not depend on the assumptions about the properties of 100% crystalline polymer. IGC data indicated that the melting temperature of PEG was 67 degrees C and crystallinity was 89.7%. DSC data indicated the same results. The experimental data was found to be not related with the chemical nature of the probes, but depended on the coating weight of PEG on the supports. PMID- 12541556 TI - A preliminary study of the quantitative structure-retention relationship of ginsenosides in normal phase thin-layer chromatography. AB - The TLC retention of ten ginsenosides has been determined on silica gel layers developed with chloroform-ethyl acetate-methanol-water (15:40:22:10, V/V) as the mobile phase. For the quantitative structure-retention relationship (QSRR) study seventeen structure parameters of the ten ginsenosides were calculated, including topologic indices and physicochemical properties. Particularly, a novel parameter "E" was presented according to the differences of the parent substances of which the ginsenosides are composed. Of these parameters, Aw, 0B and E were selected by optimization and employed in the derived multi-parameter regression equation. The relationships between the retention and the molecular structures have been discussed. PMID- 12541557 TI - [Enantiomer separation by capillary electrochromatography]. AB - As capillary electrochromatography (CEC) combines the desirable features of both high performance liquid chromatography(HPLC) and high performance capillary electrophoresis(HPCE), CEC is an attractive alternative for enantiomer separation. The applications and developments in the field of chiral drug separation by CEC are reviewed with 92 references, including different operating models and different chiral agents and stationary phases. PMID- 12541558 TI - [Advances in nonaqueous capillary electrophoresis]. AB - Advances in nonaqueous capillary electrophoresis are reviewed with 71 references in this paper. Capillary electrophoresis is generally performed in aqueous buffer. In fact, it can provide some advantages to use organic solvent as a separation medium instead of water. The choice of organic solvent and electrolyte, the detection mode and solute-additive interactions are summarized. Furthermore, the separation of inorganic ions, neural compounds, organic acids, pharmaceuticals, metabolites and chiral substance is also described. PMID- 12541559 TI - [Preparation and evaluation of glycine derivatized beta-cyclodextrin bonded silica for high performance liquid chromatography]. AB - Glycine derivatized beta-cyclodextrin bonded silica (GCDS) has been prepared for high performance liquid chromatography through the reactions of beta-cyclodextrin bonded silica with tosyl chloride and glycine in sequence. The separation performance of GCDS for positional isomers, dansyl amino acids and phenylpropionic acids was investigated. It was found that nitrophenols could not be eluted with methanol as mobile phase, but could be eluted with methanol phosphate buffer solution within appropriate time, which was much different from the behavior observed on native beta-cyclodextrin bonded silica. The effect of column temperature on the enantio-separation of dansyl D,L-Phe with GCDS was examined. The results show that the enantio-separation ability of the GCDS decreases with the increase of the column temperature. Other factors such as methanol concentration and pH of the mobile phase influencing the retention behavior of solutes on the GCDS are also discussed. PMID- 12541560 TI - [Simultaneous determination of magnolol and honokiol in serum and urine by high performance liquid chromatography]. AB - A reversed-phase high performance liquid chromatographic method for simultaneous determination of magnolol and honokiol in serum and urine of rat has been established. Two drugs were determined within 15 minutes by the method on the column with spherisorb C18, by using a mobile phase consisted of methanol-water glacial acetic acid (70:30:1, V/V) at 1 mL/min, monitored at 294 nm and with a sensitivity of 0.005 AUFS. After 0.25, 1 and 8 hour of administration of the drugs, protein in serum and urine of Wistar rat was precipitated by methanol and magnolol and honokiol in acidified body fluid were determined after being extracted by a mixture of ethyl acetate and ether. Good linear relationship between concentration in serum and urine and peak area in the ranges of 0.05-2 mg/L for magnolol and 0.025-1 mg/L for honokiol was obtained. Good precision and reproducibility were found too. The average recoveries of the two drugs were 95.6% (RSD = 3.85%), 93.8% (RSD = 3.95%) in serum and 96.0% (RSD = 3.83%), 94.9% (RSD = 3.54%) for urine respectively. The lower limit of the method was 0.02 mg/L of magnolol and 0.04 mg/L of honokiol respectively. The results showed that this method is suitable for the determination of magnolol and honokiol in body fluids. PMID- 12541561 TI - [Determination of simvastatin in human plasma by high performance liquid chromatography]. AB - Simvastatin is widely used in the treatment of hypercholestero-lemia. Recently, it was reported that it is also effective in reducing lethality in coronary heart disease. A simple and sensitive reversed-phase liquid chromatographic method has been developed and validated for the analysis of simvastatin in human plasma. Simvastatin and internal standard lovastatin in plasma were extracted with cyclohexane-dichloromethane (3.5:1, V/V), and then measured by HPLC using a Lichrospher C18 column as stationary phase and an acetonitrile-water (70:30, V/V) mixture as mobile phase. The flow rate was 1.2 mL/min. Simvastatin was quantified by UV at 237 nm. The calibration curve showed a good linearity in the mass concentration range of 0.25-50.0 micrograms/L. The regression equation is: Y = 0.0364 rho + 0.00583, r = 0.9998. Intra-day and inter-day coefficients of variation of assay for simvastatin in plasma < 7.94% and < 8.58%, respectively. The recoveries of simvastatin were > 93.3%. The method has been used to determine simvastatin in plasma from 12 healthy male volunteers. PMID- 12541562 TI - [Determination of activity of bone alkaline phosphatase in whole serum by high performance liquid chromatography]. AB - An analytical method has been established for the determination of bone alkaline phosphatase (BALP) activity using high performance liquid chromatography(HPLC). The stainless steel column was 22 cm x 4.6 mm i.d. packed with totally porous, spherical silica particles. A solution of methanol and 0.01 mol/L CH3COOH(30/70, V/V) adjusted to pH 4.8 was employed as the mobile phase. The flow rate was 0.8 mL/min. Chromatography was performed with ultraviolet detector at 275 nm. Linear calibration curve for BALP was measured within the range of 8-200 mg/L with correlation coefficient of 0.996. The lowest detection limit was 3 ng. The HPLC method described is simple, rapid and reliable and suitable for clinical monitoring. PMID- 12541563 TI - [Studies on response characteristics of Cl, Br, I of microwave plasma torch atomic emission detector for gas chromatography]. AB - The present paper deals with a microwave plasma torch atomic emission detector for gas chromatography. Argon is used as support gas, carrier gas and make-up gas. The effect of oxygen scavenger gas on the detection performance for chlorine, bromine and iodine is discussed. Detection limits, dynamic ranges, relative standard deviations and response characteristics of GC-MPT-AED for chlorine, bromine and iodine in organic compounds were studied. The results are favorable in comparing with GC-ICP-AED. PMID- 12541564 TI - [Analysis of heavy oil fractions in high-temperature coal tar by capillary gas chromatography/fourier transform infrared spectrometry]. AB - Capillary gas chromatography-Fourier transform infrared spectrometry-gas chromatography retention indices (GC/FTIR-RI), combined with the linear relationship between retention value and boiling points for polycyclic aromatic hydrocarbons, performed the determination of the composition of heavy oil fractions in high-temperature coal tar. More than 70 and 61 compounds were separated and identified in heavy oil fractions F7(300-330 degrees C) and F8(330 360 degrees C) respectively. The results are useful in coal tar distillation processing and pollution monitoring when such compounds are present. PMID- 12541565 TI - [Study on mixed stationary phase composed of 2,3,6-tributyl-beta-cyclodextrin and AgNO3 for capillary gas chromatography]. AB - A new mixed gas chromatographic stationary phase composed of 2,3,6-tributyl-beta cyclodextrin and AgNO3(in polyethylene glycol 400) was synthesized for the first time. It was coated on fused silica capillary column. Excellent selectivity was obtained for the separation of seven enantiomeric pairs and four groups of positional isomers of aromatic compounds. The recognition and separation mechanisms were discussed by measuring enthalpy, entropy, free energy and the difference in enthalpy and entropy of these compounds. Results were compared with those obtained on 2,3,6-tributyl-beta-cyclodextrin stationary phase. The results showed that the mixed stationary phase had positive synergistic effects. PMID- 12541566 TI - [A study on the chemical components of essential oil of oak moss concrete by gas chromatography/mass spectrometry]. AB - The essential oil of oak moss concrete was extracted by volatile oil content equipment. The chemical compositions and their relative contents were analyzed by GC and GC/MS. A Supelco-5 fused silica capillary column (30 m x 0.32 mm i.d.; 0.25 micron thickness) and a flame ionization detector (FID) were employed in GC analysis. The temperature program included temperature increase of 4 degrees C/min from 50 degrees C to 250 degrees C, and a 10 min isothermal period at 250 degrees C. Mass spectra were obtained by electron impact at 70 eV and a source temperature of 170 degrees C. Twenty-four volatile compounds of oak mass concrete were identified, which comprised more than 83% of volatile fraction. The major components were diethyl phthalate, alpha-terpineol, cedrane and linalool. PMID- 12541567 TI - [Determination of ibandronate by high performance ion exchange chromatography]. AB - A method for the determination of Ibandronate by high performance ion exchange chromatography is reported. Ibandronate can form a complex compound with the copper ion added in the mobile phase. The complex can be detected at UV 240 nm. The effects of pH of mobile phase and concentration of methanol to the retention time were studied. Concentration of 10% methanol and pH 2.5 were suitable for the detection in less than 5 min. The flow rate was 1.0 mL/min. The relative standard deviation of peak area was 0.8%. The linear correlation was observed from 300 mg/L to 1,500 mg/L of Ibandronate (r = 0.9989). The lowest detectable concentration of Ibandronate was 6 mg/L. The method is simple, rapid and reproducible. PMID- 12541568 TI - [Studies on the effect of column temperature on gel permeation chromatography]. AB - By experiments, we found that the effect of temperature on column efficiency increased with the increase of relative molecular mass. The elution volume was relevant to the temperature, while the selectivity was not. Besides, the resolution for macromolecules was improved significantly by increasing temperature while not for small molecules. Further, the same results were obtained at different temperatures in certain range. PMID- 12541569 TI - [Determination of inositol by reversed-phase high performance liquid chromatography]. AB - A new method for determination of inositol by reversed-phase high performance liquid chromatography was studied. The method used a Shim-Pack CLC-C18 column, acetonitrile-water (10:90, V/V) as the mobile phase at a flow rate of 1.0 mL/min with detection wavelength at 196 nm. The RSD of the results was 0.8%. It is simple, rapid, accurate, and useful for quantitative determination of inositol product. PMID- 12541570 TI - [Determination of berberine in decocted liquid from shenshu granules with water by reversed-phase liquid chromatography]. AB - A reversed-phase liquid chromatographic method has been developed and validated for the analysis of berberine in decocted liquid from Shenshu granules with water. The drug was extracted with supersonic wave and then measured by HPLC using a Hypersil C18 column and an acetonitrile -0.04 mol/L H3PO4(42:58, V/V) as mobile phase. Berberin was quantified by ultraviolet absorbance at 349 nm. The method proved to be linear in the range of 1.2-19.2 mg/L. The lower limit of detection of berberin was 0.6 mg/L. Intra-day and inter-day coefficients of variation of assay for berberin were 0.6%-3.5%(n = 5) and 5.3%-6.5% (n = 5), respectively. The recoveries of the method were 89.1%-91.4%. PMID- 12541571 TI - [Quantitative determination of chlorogenic acid in eucommia leaf by high performance liquid chromatography]. AB - An HPLC method for the quantitative determination of chlorogenic acid in Eucommia leaf was investigated. The sample was extracted with methanol solution containing 5% fomic acid and then analyzed by HPLC. The operating conditions were Shim-pack CLC-ODS column (6.0 mm i.d. x 150 mm, 5 microns) at 40 degrees C, mobile phase of methanol-0.01 mol/L KH2PO4(25:75, V/V; pH 3.9) at 1 mL/min and UV detection at 328 nm. Mercaptopurine was selected as the internal standard. There was a good linear relationship between the response (area ratio of chlorogenic acid to mercaptopurine) and the concentration of sample in the range of 25-300 mg/L for chlorogenic acid. The corresponding regression equation was Y = 8.0368X + 0.001275, r = 0.9999. The average recovery was 99.89% and RSD was 1.45%(n = 6). The results showed that this method is simple, specific and accurate. PMID- 12541572 TI - [Determination of inositol by chromatography]. AB - Inositol was acetylated to inositol hexacetate and determined by GC. The content of inositol was got. The gas chromatographic conditions were as follows: 2 m x 3 mm i.d. column packed with 10% SE-30/201 Diatoport PAW(60-80 mesh) was kept at 240 degrees C. The temperature of both injector and FID was 260 degrees C. The flowrate of N2 carrier gas was 40 mL/min. The sample was analyzed by this method with a coefficient of variation of 0.15% and a recovery of 98.2%-104.3%. PMID- 12541573 TI - [Gas chromatographic determination of camphor, menthol, methyl salicylate, thymol in JEIL COOL PAP]. AB - The contents of camphor, menthol, methyl salicylate and thymol in JEIL COOL PAP were determined with gas chromatography by using a stainless steel column (2 m x 3 mm i.d.) packed with 15% DEGS, Chromosorb W (AW-DMCS) 80-100 mesh. Temperature programming was from 70 degrees C to 180 degrees C. The quantitative determination was performed with diphenyl as the internal standard. The internal standard method showed good linearity (r = 0.9995-0.9999). The average recoveries were 99.63% (camphor), 99.83% (menthol), 100.0% (methyl salicylate) and 100.4% (thymol). PMID- 12541574 TI - [The determination of cantharidin in AIDI liquid injection by gas chromatography]. AB - A method for the determination of cantharidin in AIDI liquid injection with gas chromatography is described. The cantharidin was extracted with dichloromethane and then determined by gas chromatography with OV-17 packed column and FID detector. Its distribution constant between dichloromethane and water was 480. The extraction recoveries of cantharidin were 99%-102%. The calibration curve was Y = -113 + 35,900X, r = 0.9999 and the detection limit was 0.02 mg/L in liquid injection. As the results showed, the method is simple, rapid, sensitive and precise for determining cantharidin in AIDI liquid injection. PMID- 12541575 TI - [Determination of dimethyldiphenylurea in dibase-propellant by gas chromatography]. AB - By using gas chromatography external standard method, dimethyldiphenylurea in the storage propellant was separated and determined. Being compared with the conventional bromination method, this one is more rapid, precise and reproducible. Moreover, it can provide reliable data for evaluationg the quality grade of the propellant. PMID- 12541577 TI - [Measurement for relative molecular mass distribution of lignin dispersants]. AB - Gel chromatography was employed to measure relative molecular mass distribution of lignin dispersants. The results show that relative molecular mass distribution of Reax-85A(I) and M-9(II) are respectively to be from 500 to 35,000 and from 1,000 to 50,000. The fractions separated from (I) and (II) by thin layer chromatography were also investigated through gel chromatography to prove that these fractions were mainly separated depending on relative molecular mass. PMID- 12541576 TI - [Determination of volatile halohydrocarbons in drinking water by gas chromatography with direct aqueous injection]. AB - The volatile CHCl3, CCl4, CHBrCl2, CHBr2Cl, CHBr3 in drinking water have been determined by wide bore capillary gas chromatography/electron capture detector(GC/ECD) with direct aqueous injection method. The method is simple, fast and highly sensitive. It has better accuracy and precision. When the average mass concentration of five halohydrocarbons ranged from 9.03 to 20.19 micrograms/L, the relative deviations ranged from 5.7% to 9.1%, RSD ranged from 2.9% to 7.5%. The standard deviations ranged from 0.3 to 0.9. Correlation coefficients were 0.998 and 0.999. The detection limits ranged from 0.1 to 0.7 microgram/L and were lower than those in other work when the temperature of ECD was 350 degrees C. PMID- 12541578 TI - [Separation and determination of baicalin in prescriptions containing Scutellaria baicalensis Georgi by HPCE]. AB - Qingkailing Oral Tonic and Shuanghuanglian Oral Tonic are two kinds of complex formlae of Chinese traditional patent medicine, and both contain Scutellaria baicalensis Georgi. Baicalin is referred to the main active component with functions of dispelling wind-heat to reduce fever and detoxicating, and takes effect on uppering respiratory tract infection and tonsillities. In this work a high performance capillary electrophoresis (HPCE) method was established for separation and quantitative analysis of baicalin in prescriptions containing Scutellaria baicalensis Georgi, based on the mode of capillary zone electrophoresis. Electrophoretic conditions are as follows: uncoated fused silica capillary (34.8/39.5 cm x 50 microns i.d.), 40 mmol/L borax buffer(pH 9.0), applied voltage 17 kV, polarity anode (inlet) to cathode (outlet), temperature 25 degrees C, detection wavelength 285 nm, injection 68.95 kPa x s. Under the optimum conditions, baicalin was separated successfully from other components within 15 minutes. The relative peak area of baicalin increased linearly with the increase of its concentration in the range of 10-640 mg/L. The recoveries were (100.31 +/- 1.98)% for Qingkailing Oral Tonic and (100.60 +/- 2.36)% for Shuanghuanglian Oral Tonic. The analytical results demonstrate the method is simple, rapid and well reproducible, and can be used as a reliable tool for the quality control of Chinese traditional medicines containing Scutellaria baicalensis Georgi. PMID- 12541579 TI - [Study on the determination of trace ions in the presence of high concentration of matrix by simplified column-switching ion chromatography]. AB - A simplified column-switching technique for the determination of trace ions in the presence of high concentration of matrix is established. A binary eluent mechanism is developed to study the effect of matrix on the shift of the analyte retention time. According to this mechanism, eluent of high concentration is preferred to suppress the peak shift of trace analyte caused by large amount of matrix. Since the peak shift has been effectively suppressed, the procedures for determining the column-switching time windows were greatly simplified. According to the different samples, different strategies should be employed in this technique. In this work, three typical samples were analyzed by this technique and the strategies for these three samples were studied to show the principles of choosing the proper separation conditions. Results showed that by this simplified technique analysis for trace analytes in the presence of complicated matrix was accurate and precise. The successful use of this method to the determination of trace bromate in drinking water, and nitrate, ammonium and iodide in sea water indicates that the application of this versatile technique to quantify analytes of low concentration in a wide range of sample matrices is promising. PMID- 12541580 TI - [Analysis of multi-wavelength overlapping chromatographic data by orthogonal projection resolution]. AB - Orthogonal projection resolution(OPR) has been proved to be a high performance chromatographic data processing technique. In this paper, it is applied to the resolution of multi-wavelength chromatographic overlapping peaks. When the highest number of degree of overlapping in chromatogram is less than or equal to the number of wavelengths used, a real resolved chromatogram can be completely obtained from multi-wavelength chromatographic overlapping peaks by this method. When based on dual wavelength chromatographic data analysis, a new method, namely dual wavelength characteristic information analysis (DWCI), used for the base line correction, determination of number of components and region of pure components signal. Analysis of overlapping cases of every components in the overlapping chromatogram is also proposed. DWCI has been successfully used for the analysis of overlapping chromatogram with dual peaks of three components and with single peak of two components. But based on single wavelength chromatographic data, the analysis of these overlapping peaks is a very difficult and complicated problem. PMID- 12541581 TI - [Separation model of small molecules with SEC columns and micellar mobile phase]. AB - Small alkanol molecules were successfully separated with SEC method in combination with a micellar mobile phase. The separation model has been given in this system. The separation is based on the differential migration of the micelle and the surrounding water and the differential distribution of the solute between the micelle and aqueous phase, therefore, the separation mechanism is very similar to that of micella electrokinetic chromatography (MEKC). The retention volume of solute is related to the micellar concentration by theoretical treatment, through which the solute distribution coefficient between micelle and aqueous phase could be obtained. Two different SEC columns were employed for separation of some alkanols and the theoretical model is supported by the experimental results. The distribution coefficients obtained were in good agreement for these two columns. PMID- 12541582 TI - [Research on the separation of limonoid glucosides by reversed-phase preparative high performance liquid chromatography]. AB - Obacunone-17-beta-D-glucopyranoside (OG) was isolated from the seeds of Citrus Sinensis Osbeck by using solvent extraction, classical polymer adsorption column separation and weak base anion ion-exchange separation, OG was finally purified by C18 reversed-phase preparative high performance liquid chromatography and was identified by thin-layer chromatography. The purity of OG was analyzed by analytical reversed-phase HPLC. At last the structure of OG was determined by 1H and 13C nuclear magnetic resonance spectrometry (NMR). In this work, the conditions of the reversed-phase preparative HPLC technique to purify limonoid glucosides was optimized. The reversed-phase preparative HPLC on a C18 column with a mobile phase of acidic acetonitrile-water (about 0.2% TFA, V/V) at pH 3 enabled the baseline separation of limonoid glucosides in the extract. The results show that OG is the predominant limonoid glucoside in the seeds of Citrus Sinensis Osbeck and nomilin glucoside is the second one. The results also show that the classical polymer adsorption column separation and weak base anion ion exchange separation are effective for purifying limonoid glucosides. PMID- 12541583 TI - [Preparation of novel chiral stationary phases for ligand exchange chromatography and their applications]. AB - Two novel chiral stationary phases for ligand exchange chromatography were prepared by synthesizing 2-(2-hydroxyl-3-octoxy)propyl-(S)-1, 2, 3, 4-tetrahydro 3-isoquinolinecarboxylic acid (C8-(S)-THIQCA) and 2-(2-hydroxyl-3-dodecoxy)propyl (S)-1, 2, 3, 4-tetrahydro-3-isoquinolinecarboxylic acid (C12-(S)-THIQCA) as chiral selectors, and coating them on ODS bonded stationary phase. Chromatographic resolutions of some DL-amino acids were achieved on the chiral stationary phases by using 1 mmol/L NH4Cl with 0.2 mmol/L Cu(Ac)2 solution as mobile phase and detection at 254 nm. Ability of chiral resolution of the DL amino acids through the stationary phases was compared and the effect of cupric ion on resolutions of the DL-amino acids was examined. The elution order of D isomer before L-isomer was observed for all the DL-amino acids resolved, except DL-Val. For DL-Pro, DL-Val and DL-Leu the elution order through the chiral phases was different from that through the chiral ligand exchange phases prepared from L Pro or L-hydroxyl proline with five membered ring structure. PMID- 12541584 TI - [Sample preconcentration technique in capillary electrophoresis]. AB - The methods of improving the detection sensitivity in analysis of trace level components by sample preconcentration technique including sample stacking, field amplified sample injection, isotachophoresis etc. are reviewed, and 67 references are cited. PMID- 12541585 TI - [The latest applications of ion chromatography in food analysis]. AB - The latest applications of ion chromatography in food analysis is reviewed with 60 references, and the development trend in this aspect is also discussed. PMID- 12541586 TI - [Studies on quantitative chromatographic retention-structure relationships]. AB - Using quantum and molecular mechanics methods, 48 structural descriptors of alkylbenzene compounds such as atomic partial charge, dipole moment, molecular area, molecular volume etc. were calculated. GFA regression methods were applied to build a quantitative chromatographic retention-structure relationships equation. The equation have good fitness and strong predictivity. Principle structural factors that effect chromatographic retention of alkylbenzenes were discussed. PMID- 12541587 TI - [Investigation on the changing regularity of the relationship between retention index and temperature of the GDX series stationary phases]. AB - A systematical measure of the polarity scale of the Chinese porous polymer beads of this series of stationary phase has been carried out, including the McReynolds constant at 120 degrees C by using squalane as non-polar stationary phase and the relative constant to GDX-103 at 180 degrees C. Based on the above work, two kinds stationary phases GDX-101 and GDX-103 were selected, on which the changing regularity of the relationship between retention index and temperature of the five "probes" of stationary polarity determination, including benzene, butanol, 1 nitropropane, 2-pentanone and pyridine were investigated. At the same time, the changing regularity with temperature of the relative McReynolds constant delta I of GDX-101 to GDX-103 was studied. It is found that Ip-theta c curve and delta I theta c curve all showed a turning point at the temperature of 180 degrees C and that there was a maximum value of delta I at the temperature of 180 degrees C. So we think that the retention mechanism of this kind of stationary phase changed at 180 degrees C. PMID- 12541588 TI - [Investigation of a gas chromatographic column system for the on-line analysis of gaseous components in de-propane tower of pyrolysis equipment]. AB - Multi-dimensional gas chromatograph has become an important process analyzer due to the advantages of high resolution and fast speed. According to the production requirement, a gas chromatographic column switching system has been investigated for the on-line analysis of gaseous components from high-pressure and lower pressure de-propane towers of pyrolysis equipment. By using two different injection times on three injectors, and fore-flush and back-flush techniques, C2 hydrocarbons, propane, propene, methylacetylene, propadiene and C4-hydrocarbons can be separated on 7 columns in 7 minutes. The practical application showed the developed column system is suitable for the on-line monitoring of the production process. PMID- 12541589 TI - [Studies on shape selective isopropylation products of coal tar naphthalene by gas chromatography/mass spectrometry]. AB - Shape selective isopropylation products of coal tar naphthalene over different zeolite catalysts have been studied by capillary gas chromatography/mass spectrometry (CGC/MS) combined with computerized library search and manual interpretations of mass spectra. The analytical results have shown that products mainly consisted of some monosubstituted, disubstituted and a few trisubstituted naphthalenes. Except the low activity of HZSM-5 catalyst, all other zeolite catalysts have higher activity. HY and H beta catalytic activities were the highest, and their naphthalene conversions attained to 79.12 and 55.79 percent respectively, but their selectivities for 2-isopropylnaphthalene (beta-IPN), 2, 6 diisopropylnaphthalene (2, 6-DIPN) and 2, 7-diisopropylnaphthalene (2, 7-DIPN) were too low. ZM catalysts had good activity and much better selectivity, especially for ZM4-3, of which its 2,6-DIPN/2,7-DIPN ratio was 2.97. Therefore, some zeolite catalysts are recommended for shape selective isopropylation of coal tar naphthalene. PMID- 12541590 TI - [Determination of alkyl alcohol polyoxyethylene ether by high temperature gas chromatography]. AB - High temperature GC method has been developed for the separation and determination of alkyl alcohol polyoxyethylene ether (AEO). These AEO samples were separated on high temperature Al-coated fused-silica capillary column (0.1 micron bonded methyl silicon stationary phase, 25 m x 0.25 mm i.d.). The components of AEO sample were identified by GC/MS. The free alkyl alcohol and ethoxymer distribution of polyoxyethylene of AEO sample were determined by normalization method. The FID responses of typical components of AEO sample were determined, and their relative deviations were less than 4.1%. The recoveries of the free alkyl alcohol ranged from 96.5% to 98.1%. The relative standard deviations were less than 1.9%. In comparing with previous methods, this method is simple, fast and more reproducible. PMID- 12541591 TI - [Determination of phospholipid in the peroxisomal membrane of rat liver by high performance liquid chromatography]. AB - A method was employed for the determination of phospholipid in membrane of peroxisomes by high performance liquid chromatography. The separation was performed on a mu-Porasil Si60 column(250 mm x 4.6 mm i.d.) by gradient elution. The phospholipids were detected at 205 nm. The limits of detection for cardiolipin, phosphatidylinositol, phosphatidylserine, phosphatidylethanolamine, phosphatidylcholine and sphingomyelin were 12, 8, 23, 4, 23 and 7 ng respectively and the recoveries were between 84% and 103%. The linear relationship of calibration curves was good (r > 0.91). The phospholipids in peroxisomal membrane of 10 SD rat liver were determined and the results are satisfactory. PMID- 12541592 TI - [Study on the residual dynamics of flumioxazin in soybean and soil]. AB - The residual dynamics of flumioxazin in soil and soybean were determined with HPLC, using a Zorbax SIL column and methylene dichloride-methanol (99.5:0.5, V/V) as mobile phase. A method for pretreatment of samples and analysis was established. Recoveries of flumioxazin in soil, green soybean and soybean were 84.82%-85.63%, 88.79%-94.00% and 93.70%-94.48%, respectively. Flumioxazin degrades relatively slowly and its half-life in soil was 10.0-10.5 days, but in the soil, green soybean and soybean residual flumioxazin was not detected. The results showed that the residual pollution of flumioxazin is negligible, so the application of flumioxazin is safe for the soil and soybean. PMID- 12541593 TI - [Enantiomeric separation of N-FMOC amino acids with sulfobutyl ether-beta cyclodextrin as chiral additive by capillary electrokinetic chromatography]. AB - Enantiomeric separation of N-FMOC amino acids by capillary electrophoresis with an anionic chiral resolving agent, sulfobutyl ether-beta-cyclodextrin (SBE-beta CD) with average substitution degree of 3.8, employed as chiral additive was studied. Chiral separation was achieved with the mode of electrokinetic chromatography due to the electrophoretic mobility and chiral complexing ability of the chiral additive. The effects of pH of background electrolyte, the concentration of sulfobutyl ether-beta-cyclodextrin on enantiomeric separations were investigated. The enantiomeric separations of 8 N-FMOC amino acids were accomplished, among which 5 N-FMOC amino acids could be on baseline separated. PMID- 12541594 TI - [Study on the separation of proteins in human serum by high performance capillary zone electrophoresis]. AB - A high performance capillary zone electrophoretic method has been developed for the separation of proteins in human serum. Sera were diluted with a buffer (0.05 mol/L boric acid, pH 8.80), and then introduced hydrodynamically into a 37 cm (32 cm to detector) x 50 microns i.d. fused-silica capillary, electrophoresed for 12 min using a boric buffer solution (0.10 mol/L, pH 9.35; containing 4 g/L of polyethylene glycol 8000), and detected at the wavelength of 200 nm. The method is simple, rapid and repeatable. PMID- 12541595 TI - [Effect of sample solvents on retention in packed column supercritical fluid chromatography]. AB - Factors affecting the retention in supercritical fluid chromatography(SFC) on C18 and NH2 packed columns have been studied. These factors include the sampling solvent, continuous injection, stationary phase type and concentration of modifier. When supercritical carbon dioxide or low concentration(less than 0.5%) of methanol is employed as the mobile phase, retention time increases with the polarity of the solvent on NH2 column, while it keeps almost constant on C18 column. Methanol will cover the active sites on the surface of the bonded phase, forming the "pseudo" stationary phase, which will lower the interaction between the analytes and the stationary phase, hence the shorter will be the retention time. The hydrogen bond between stationary phase (including the amino group and the remaining hydroxyl group) and methanol is the most obvious. So it's quite more evident on NH2 column than on C18 column that the delaying effect of the solvent, with methanol, acetone and chloroform in decreasing sequence, encourages the elution of the analytes during continuous injections. The elution will recover to the first injection after adequate interval has passed, i.e., more than 10 column volumes. When the methanol concentration is greater than 1.0% in carbon dioxide, the effect decreases and becomes negligible. The solvent effect is significant to the reasonable injection and reproducible chromatographic data in packed column SFC. PMID- 12541596 TI - [Capillary gas chromatographic determination of residual benzene, toluene and DMF in acrylic fabrics]. AB - A simple method for the analysis of residual benzene, toluene and N,N-dimethyl formamide (DMF) in acrylic fabrics by wide-bore capillary gas chromatography is presented. The sample was extracted by carbon disulfide (CS2) for an hour. The results showed that this is a rapid and sensitive method with less interferences and fairly good precision. The detection limits were 0.1, 0.1 and 2.0 ng for benzene, toluene and DMF respectively. The recoveries were 93.3%-101.8% and coefficients of variation were 2.0%-3.4%. PMID- 12541597 TI - [Study on the direct determination of auxins by gas chromatography]. AB - A method for the direct determination of auxins, indole acetic acid (IAA), indole butyric acid (IBA) and naphthylacetic acid (NAA), by gas chromatography with wide bore capillary column is described. The gas chromatographic conditions were as follows: HP-1 capillary column, 5 m x 0.53 mm i.d., 2.65 microns film, the column temperature 180 degrees C, the temperature of injector 250 degrees C, the temperature of FID detector 260 degrees C, carrier gas: N2 20 mL/min and dibutyl phthalate was used as internal standard. All components and internal standard were separated in 3 min. The relative standard deviations were from 0.61% to 1.14%. The method is simple, rapid, sensitive, good reproducible and satisfactory. PMID- 12541598 TI - [Determination of the butyl 2,4-dichlorophenoxyacetate in the workshop by gas chromatography]. AB - The new method for determining the content of 2,4-dichlorophenoxyacetic acid butyl ester in the workshop by using gas chromatography is presented. The experiments for precision and efficiency have been performed. The range of determination is 0.05-2.5 mg/m3. The minimum detectable mass concentration is 25 micrograms/m3. The method meets the requirement of the "monitoring and researching standard for toxic substances", and shows that it can be applied in routine analysis. PMID- 12541599 TI - [Study on the packings of affinity chromatography for the separation of urokinase]. AB - Two kinds of affinity chromatographic packings for the separation of urokinase were synthesized by coupling of p-amino benzamidine (p-ABZ) to commercially available sepharose and polyepoxypropyl methacrylate (PEPMA). Then they were applied for separating crude urokinase. It was found that the average recovery of bioactivity on sepharose was higher than that on PEPMA, resulting in 108.3% and 43.4% respectively. The high rigidity of PEPMA permits fast flow of protein solutions and operation by higher-pressure affinity chromatography. The average purification times were 36.9 folds for PEPMA column and nine-folds for Sepharose column. The purification of crude urokinase described in this paper demonstrates that PEPMA column is effective for purifying biological products in a large scale. PMID- 12541600 TI - [Studies on the active constituents of sichuan liang shan rhododendron L. I. Determination of two flavonoids in leaves, extracts and jinjuan liquors]. AB - A reversed-phase HPLC method for the separation and determination of the two flavonoids: quercetin and kaempferol is described. The column was 25 cm x 4.6 mm i.d., packed with Zorbax SB-C18(5 microns). The mobile phase was a mixture of MeOH-H2O (60:40, V/V). The flow rate was 0.7 mL/min. UV detection was performed at 360 nm. External standard was used and the calibration curves showed good linearity over the range of 6.0-100 mg/L, r = 0.9995-0.9998, and detection limits were 3.3-6.2 mg/L. The recoveries were 96.0% and 97.3% for quercetin and kaempferol respectively. The relative standard deviations of the method were 2.1% 4.5%. The method is simple, fast, sensitive and accurate and can be applied to the analysis of quercetin and kaempferol. It provides a scientific basis for industrial production and quality control of Jinjuan liquor preparations for clinical uses. PMID- 12541601 TI - [Simultaneous analysis of low concentrations of glucose, ethanol and glycerol by high performance liquid chromatographic method]. AB - A method based on high performance liquid chromatography (HPLC) for the analysis of glucose, ethanol and glycerol simultaneously is presented. The baseline separation was achieved at room temperature (25-30 degrees C) on a Beckman mu Spherogel carbohydrate column (6.5 mm i.d. x 30 cm) with H2SO4-H2O(0.5:1,000, V/V) as mobile phase at a flow rate of 1.000 mL/min. All the substances were detected with the Beckman 186 Refractive Index Detector. Under these conditions, distinct peaks of glucose, ethanol and glycerol were resolved within 15 min. The retention time in the particular condition was (6.04 +/- 0.04) min for glucose, (13.36 +/- 0.08) min for ethanol and (8.72 +/- 0.05) min for glycerol. The detection limit for glucose, ethanol and glycerol were found to be 10(-5) g, 10( 4) g and 10(-5) g respectively. A series of experiments have been performed to investigate the glucose consumption and the ethanol, glycerol production in the random growth of Saccharomyces cerevisiae with the method above. Only 0.5 mL sample was needed each time. In 30 hours after inoculation, the cell density increased from 10(6) mL-1 to 10(7) mL-1. It was found that the glucose concentration decreased approximately linearly after 8 hours, and ethanol and glycerol was detected after the delay period as expected. The results show that the method proposed is rapid and convenient with enough accuracy. The method may be used to investigate the metabolic behavior of yeast in the fermentation process at the early stage or detect the slight changes of concentrations of some objective substances in particular culture experiments. PMID- 12541602 TI - [RP-HPLC determination of MBAMT and its decomposition product]. AB - This paper presents a method of reversed-phase high performance liquid chromatography to determine N,N-methylene-bi-(2-amino-5-mercapto-1,3,4 thiodiazole) (MBAMT) and its decomposition product 2-amino-5-mercapto-1,3,4 thiodiazole (AMT). Recoveries of MBAMT and AMT were 91% and 94% respectively. The detectable limits of them in water samples were 7.19 nmol/L for MBAMT and 2.87 nmol/L for AMT. The resolution between the peaks of MBAMT and AMT was 2.5. The column efficiency and the sensitivity of this method were significantly better in comparison with normal phase chromatography. PMID- 12541603 TI - [Determination of rufloxacin in human plasma by high performance liquid chromatography]. AB - A high performance liquid chromatographic method has been developed for the determination of rufloxacin in human plasma. Rufloxacin was extracted from plasma with dichloromethane for three times. It was chromatographed on an Ultrasphere ODS column with Pefloxacin as internal standard with a mobile phase consisting of methanol-tetrabutylammonium bromide-triethanolamine (32:68:0.5, V/V) adjusted to pH 2.8 with orthophosphoric acid. The flow rate was 1.2 mL/min and the monitoring wavelength was 295 nm. The calibration curve was linear from 0.1 to 10 mg/L of plasma. The detection limit of rufloxacin was 0.05 mg/L for plasma and the recovery was (97.7 +/- 2.1)%. The intra-day RSD and inter-day RSD were 2.33% and 3.38% respectively. The method is simple, rapid, accurate and can be used to determine the rufloxacin concentration in plasma and for pharmacokinetic study. PMID- 12541604 TI - [Determination of sotalol hydrochloride by reversed-phase high performance liquid chromatography]. AB - An RP-HPLC method for determination and detection of sotalol hydrochloride is described. The baseline separation was achieved on ODS column within 15 minutes by using 0.1% HAc-acetonitrile (80:20, V/V) as mobile phase at a flow rate 1.5 mL/min, and the wavelength was set at 227 nm. The linear range was 5-45 mg/L (r = 0.9991) and the limit of detection was 1 mg/L(S/N > 3). The intra-day and inter day RSDs were 0.20% and 0.93% respectively. PMID- 12541605 TI - [Analysis of imidacloprid and damanlin by high performance liquid chromatography]. AB - A new method for determination of imidacloprid and damanlin by using HPLC was established. It was carried out on a mu Bondapak C18 column (3.9 mm i.d. x 300 mm) with ACN-MeOH-H2O (60:10:30, V/V) as eluent and detected at 240 nm using photodiode array detector. The results of imidacloprid and damanlin were 3.71% and 15.65% respectively. The standard deviations for imidacloprid and damanlin were 0.88% and 0.76% respectively. PMID- 12541606 TI - [Synthesis and application of sulfonated beta-cyclodextrins as chiral additives in capillary electrophoresis]. AB - Three new kinds of the sodium salt of sulfonated beta-cyclodextrins with different degrees of substitution were synthesized and characterized. A simple synthesis method was employed by direct sulfonating reaction with concentrated sulfuric acid. Sulfonated beta-cyclodextrins were used as chiral resolving agents for the capillary electrophoretic separation of enantiomers in high pH and low pH background electrolytes. In different electrophoretic polarity mode, the effects of the type and concentration of sulfonated beta-cyclodextrin were investigated. Sulfonated beta-cyclodextrins were proved to be strong complexing agents for basic and neutral analytes. PMID- 12541607 TI - [Determination of O6-methylguanine by high performance capillary zone electrophoresis]. AB - In this paper, a method to determine O6-methylguanine in urine by high performance capillary zone electrophoresis has been established. O6-methylguanine was directly introduced into the capillary employing sodium borate at pH 9.0 as buffer. The capillary used was 75 microns i.d. x 57 cm. The calibration curve showed good linearity, r = 0.9987, CVs were 1.51% for intra-day and 1.93% for inter-day. The average recovery was 98.2%. In conclusion, this method is a simple, rapid, precise and reliable technique for O6-methylguanine measurement in urine. PMID- 12541608 TI - [Chromatographic analysis on tryptic hydrolysis of whole casein]. AB - Tryptic hydrolysis of whole casein was investigated by high performance size exclusion chromatography (HPSEC) in combination with the degree of hydrolysis (DH). In terms of HPSEC chromatograms obtained at different DH values, the complex process of enzymatic reaction and the relative molecular mass distribution of multiple hydrolysates were quantitatively characterized. Based on the information of casein micelle structure, the possible reaction mechanism was deduced from a series of chromatograms. Being taken into account the primary structure of whole casein and the target amino acid of trypsin, the distribution of theoretical peptides were accurately calculated by determining the split sites of complete enzymatic hydrolysis. According to the relationship between retention time and relative molecular mass, the corresponding HPSEC absorption peaks of active peptides in hydrolysates were identified, and caseinophosphopeptides sequences were also characterized. PMID- 12541609 TI - [Relationship study between the retention behavior on diverse gas chromatographic stationary phases and molecular structure information connectivity indices of PCDFs]. AB - Based on the molecular valence connectivity index, atomic characteristic value(valence delta) delta iH is defined as: delta iH = mi.Zi.(Zi-hi).(8 Ni)(hi/(2.ni))/ [4.(Li + pi).(ni-1)]. Molecular valence connectivity index n chi H (n = 1,2,...,m) of atomic characteristic are set up with the delta iH:n chi H = sigma (delta iH. delta jH. delta kH..... delta mH)-0.5 and n chi H being defined as: 1 chi H = sigma (delta iH. delta jH)-0.5, 2 chi H = sigma (delta iH. delta jH. delta kH)-0.5. The 1 chi H, 2 chi H values of 135 polychlorinated dibenzofurans (PCDFs) molecules are calculated. It is found that 1 chi H or 2 chi H or 1 chi H and 2 chi H are correlated well with the retention behavior(RI and RRT) of gas chromatography for these compounds. Twelve models, each of which is constructed by using all sample sets, with high correlation coefficient, r > 0.96, are developed for three columns (DB-5, SE-54 and OV-101). It has been demonstrated that the method possesses the advantages of easy computation and clear physical significance. PMID- 12541610 TI - [Analysis of confined crystalline behaviors of poly(styrene)-poly(ethyleneoxide) poly(styrene) triblock copolymers by inverse gas chromatography]. AB - The confined crystalline behaviors of the triblock copolymers, poly(styrene) poly(ethyleneoxide)-poly(styrene) (PS-PEO-PS), were studied by using inverse gas chromatography(IGC) probe technique, including phase-transformation of melting crystalline, crystallinity (Xc), melting temperature(Tm) and melting range of temperature. The effects of the molecule-chain length of linear alkane probes on the results are discussed. Results showed that micro-phase separation of PS-PEO PS had a greater influence on crystallization of PEO molecule-chain. Crystalline structure of PS-PEO-PS had interphase formed by some kinds of imperfect PEO crystal and amorphous PS. The molecule-chain length of linear alkane probes had no effect on the determination of melting temperature and melting range of temperature of PS-PEO-PS, but had a greater influence on the determination of crystallinity of PS-PEO-PS and investigation phase-transformation of melting crystalline. Crystallinity of PS-PEO-PS determined by IGC was decreased with the increase of molecule-chain length of linear alkane probes. By suitable shorter molecule-chain length of linear alkane probes, it was truer to reflect the existence of interphase of PS-PEO-PS and multi-phase-transformation of melting crystalline presenting in interphase. PMID- 12541611 TI - [New technologies of solid-phase microextraction]. AB - Solid-phase microextraction (SPME) is a technology with functions of sampling, extraction and concentration based on adsorption/absorption-desorption equilibrium between coating and sample. The recent development of SPME is reviewed. Studies on the in-tube-SPME technology, novel extraction probes and its coupling with other analytical technologies are summarized. PMID- 12541612 TI - Study on retention in liquid chromatography. AB - After joining the faculty at Toyohashi University of Technology (TUT) in 1978, the author has found two areas in separation sciences where microcolumn liquid chromatography (micro-LC) can be beneficial. One is the hyphenated techniques between many spectroscopic methods such as mass (MS), infrared (FT-IR) and atomic emission (ICP), and micro-LC. The other one is rather difficult, but basic and theoretical approach which deals with retention mechanism in LC. The latter project has especially been giving him a lot of scientific funs and honors in the last 20 years. On the occasion of being awarded by The Society for Chromatographic Sciences the author would like to summarize his contributions to this topic which asks us "What Is Chromatographic Retention?" PMID- 12541613 TI - [Plate theory model under non-ideal chromatography]. AB - The mobile phase in chromatography is regarded as to be composed of many continual plates, whose height is the same as that of one plate in the stationary phase. The solute in initial concentration of mobile phase is seen to be concentrated in the first plate. The solute is exchanged dynamically between the mobile phase and stationary phase. When the mobile phase passes the distance of one plate in the stationary phase, the solute cannot be immediately released from the stationary phase to mobile phase, because of the dynamic factors of chromatographic system. The solute distribution between mobile and stationary phases is controlled by two factors, i.e. the equilibrium coefficient P, which represents chromatographic thermodynamic properties, and release probability factor alpha, which represents chromatographic dynamic properties. This process is considered as non-ideal chromatography. In the view of statistics, authors studied the influences integrated of the release property factor and equilibrium coefficient factor upon chromatographic distribution, and simulated the chromatographic distribution curve by means of computer. As a result, a new distribution model has been established, the distribution equation under the non ideal chromatography was deduced and, the integrative action of the release probability and the equilibrium coefficient was revealed. The release probability factor can be regarded as an index in chromatography and the peak width can be deduced and hence the resolution can be calculated through this index. PMID- 12541614 TI - [Synthesis of a new cyclodextrin derivative and its application as CGC chiral stationary phase in determination of enantiomeric excess]. AB - A new cyclodextrin (CD) derivative, 2,6-di-O-benzyl-3-O-valeryl-beta-CD, was synthesized and characterized by 1H NMR and IR. Using the beta-CD derivative as chiral stationary phase of capillary gas chromatography, one chiral column was prepared. On this column, the enantiomeric excesses (e.e.) of 1-(2,4 dichlorophenyl) ethanol obtained by asymmetric catalytic hydrogenation of 2,4 dichloroacetophone and trans-3-propyloxiranemethanol synthesized by Sharpless epoxidation of trans-2-hexenol were determined and the catalytic reactions were evaluated. The results are satisfactory. PMID- 12541615 TI - [Pharmacokinetic studies on artemether under conditions simulated in vivo]. AB - Pharmacokinetics of artemether under conditions simulated in vivo was studied by high performance liquid chromatography. The pharmacokinetic curves of artemether and its metabolites dihydroartemisinin and furano acetate of artemether were determined and the results showed that the half-life of artemether under the conditions simulated the pH of plasma or small intestine was more than 150 min and that of stomach acid was 74.6 min. The results also showed that dihydroartemisinin and furano acetate of artemether were the main metabolites of artemether under the above conditions. Furthermore, the determination methods reported in literature were improved. PMID- 12541616 TI - [Extraction of three tanshinones from the root of Salvia miltiorrhiza Bunge by supercritical carbon dioxide fluid and their analysis with high performance liquid chromatography]. AB - The extraction of three tanshinones from the root of Salvia miltiorrhiza Bunge by supercritical carbon dioxide (SC-CO2) fluid with ethanol as co-solvent was investigated using four-factor and three-level orthogonal design method and the systematic method. The optimized conditions were established as follows: 95% ethanol was selected as a co-solvent, flow rate of co-solvent was 1.0 mL/min, extraction pressure was 20 MPa, extraction temperature was 45 degrees C, separation temperature was 35 degrees C. The sample solution was separated on a Nova-Pak C18 column (3.9 mm i.d. x 300 mm, 5 microns) with CH3OH-H2O (80:20, V/V) as the mobile phase and detected at 280 nm. The correlation coefficients were 0.9994-0.9998 and the relative standard deviations were 2.37%-3.47%. The detection limits were 0.473 mg/L, 0.184 mg/L and 0.452 mg/L for tanshinone-II A, tanshinone-I and cryptotanshinone respectively. PMID- 12541617 TI - [Determination of nanchangmycin and meilingmycin by high performance liquid chromatography]. AB - A high performance liquid chromatographic method had been established for the separation and determination of two antibiotics produced by Streptomyces nanchangensis, polyether nanchangmycin and 16-membered macrolide meilingmycin. The latter is composed of several components. The operating conditions were Waters XTerra RP18 column (3.9 mm i.d. x 150 mm, 5 microns) at 25 degrees C, mobile phase linear gradient elution of acetonitrile-water with 57:43 (volume ratio) during 0 min-30 min, 70:30 during 30 min-32 min, 80:20 during 32 min-34 min, 90:10 during 34 min-50 min at a flow rate of 0.8 mL/min, and photodiode array detector at 234 nm. This method is accurate, rapid and simple, and can be used for the integrated analysis of the two different natural compounds, polyethers and macrolides. PMID- 12541618 TI - [Determination of organic acids in fermentation broth of spiramycin by high performance liquid chromatography]. AB - A method for determining organic acids in spiramycin fermentation broth by high performance liquid chromatography is described. The operating conditions were Zorbax 300-SB C18 column (5 microns, 4.6 mm i.d. x 15 cm) at 35 degrees C, 0.01 mol/L phosphoric acid buffer solution (pH 2.32) and methanol as mobile phase at a flow rate of 0.6 mL/min and UV detection at 210 nm. The relative standard deviations were 0.33%-0.10% and the recoveries were 99.95%-100.08%. It's a simple, rapid and accurate method. PMID- 12541619 TI - [Determination of tetracyclines antibiotics by reversed-phase high performance liquid chromatography]. AB - A method for the determination of 7 tetracycline antibiotics (TCs) by reversed phase high performance liquid chromatography is described. TCs were successfully separated on a Diamonsil C18 column (250 mm x 4.6 mm i.d., 5 microns) using methanol-acetonitrile-0.01 mol/L oxalic acid (pH 2.0) (11:22:67, volume ratio) as mobile phase at a flow rate of 0.8 mL/min and detected at 267 nm within 22 min. Effects of the pH value of the mobile phase, concentration of the mobile phase buffer, elution composition and detection wavelength on the response and retention were studied. Tetracycline (TC) and oxytetracycline(OTC) in medicinal tablets were quantitated by standard added method. It has been proven that the method is fast, accurate and suitable for routine analysis. PMID- 12541620 TI - [Rapid analysis of tryptophan in serum by high performance liquid chromatography with fluorescence detection]. AB - A sensitive, rapid and specific method for the determination of tryptophan(Trp) in serum by high performance liquid chromatography with fluorescence detection (HPLC-FLD) has been established. Serum sample was precipitated by 5% perchloric acid solution and centrifuged to remove protein and then assayed by HPLC-FLD. The operating conditions were Nova-Pak C18 column (3.9 mm i.d x 150 mm, 4 microns), 5 mmol/L KH2PO4 as the mobile phase at a flow rate of 1.0 mL/min. The fluorescence detector was operated at lambda ex 254 nm and lambda em 338 nm. The method was proved to be linear in the range of 0.49 mumol/L-490.00 mumol/L with a regression coefficient of 0.9999. The minimum detection limit was 0.10 mumol/L. The recoveries were 97.2%-98.6%. The intra-batch and inter-batch RSDs were 2.0% and 3.2% respectively. The detection time of serum Trp was within 5 min after injection. The results suggest that this method is simple, fast, accurate and convenient. PMID- 12541621 TI - [Determination of ropivacaine in human serum by reversed-phase high performance liquid chromatography]. AB - A simple and rapid reversed-phase high performance liquid chromatographic method was developed for the determination of ropivacaine. The sample was treated by the addition of bupivacaine as an internal standard, extraction with dichloromethane, and drying by N2. Then the residue dissolved in the mobile phase and 40 microL were injected into the chromatographic system with a mobile phase of 0.01 mol.L-1 KH2PO4 (pH 3.0)-CH3CN(84:16, volume ratio). The detection wavelength was 210 nm. The retention times of ropivacaine and the internal standard were 5.5 min and 10.5 min respectively. The detection limit was 0.025 mg.L-1. The linear range was from 0.05 mg.L-1 to 2.50 mg.L-1 with r of 0.9997. The average recovery was 104.1% with the intra-day and inter-day RSDs of 3.42% and 6.27% respectively. The results showed that it is a simple, rapid and stable method for drug monitoring in clinical research. PMID- 12541622 TI - [Determination of trace chrysene in environmental water by solid-phase microextraction coupled with high performance liquid chromatography]. AB - A rapid method for the determination of trace chrysene in environmental water by solid-phase microextraction (SPME) coupled with high performmnce liquid chromatography (HPLC) was developed. The experimental conditions of SPME, such as extraction time, extraction temperature, effect of ion strength, desorption time, desorption solution, desorption mode and the analytical conditions of HPLC were optimized. The optimal conditions were room temperature, 1,100 r/min of stirring rate, 30 min of extraction time, 3 min of desorption time, and methanol as the desorption solution. Methanol was used as the mobile phase on a C18 reversed phase chromatographic column. The flow rate was 1 mL/min. The detection wavelength was 266 nm on a UV-Vis detector. The linear range was from 0.013 microgram/L to 3.0 micrograms/L, the detection limit was 2.7 ng/L, and the relative standard deviation (RSD) was 5.6%. The method was used for the determination of trace chrysene in tap water, mineral water, rain water and river water. The recoveries were from 103.2% to 119.3%, the RSDs were from 4.8% to 10.2%. The method is fast, convenient, sensitive, solvent-free, and suitable for the determination of trace chrysene in environmental water. PMID- 12541623 TI - [Improvement of the determination method of benzene, toluene, ethylbenzene and xylene(BTEX) in water using activated carbon fiber solid-phase microextraction/gas chromatography-mass spectrometry(GC-MS)]. AB - The methods of direct injection, carbon disulfide extraction and activated carbon fiber solid-phase microextraction/GC-MS, usually used in the determination of BTEX in water matrix, are compared and discussed. Experimental data of linearity, precision and limit of detection illustrate that the last one is better than the two other methods. This method was tested by the practical sample experiments and expected to be a simple and sensitive new method for the analysis of BTEX in water. PMID- 12541624 TI - [Determination of perfluorooctanoic acid in plasma by gas chromatography]. AB - A method for the quantitative determination of perfluorooctanoic acid in plasma by gas chromatograph-electron capture detector is described. An HP-1 capillary column (30 m x 0.25 mm i.d. x 0.25 micron) was used. Perfluorodecanoic acid as the internal standard was added to plasma sample, which was then lyophilized. The acids were derivatized to their methyl esters and then extracted into hexane. The calibration curve was linear over a range from 20.00 micrograms/L to 4.00 x 10(3) micrograms/L with a correlation coefficient of 0.9946. The average recovery was 99.2% and the RSD was 0.95%. The detection limit was 6.0 micrograms/L. The level of perfluorooctanoic acid in plasma of related occupational workers was measured. The results showed that this method is accurate and reliable. PMID- 12541625 TI - [Quantitative determination of morphine in opium powder by addition and correlation method using capillary electrophoresis]. AB - The morphine in opium powder has been quantitatively determined by addition and correlation method (ACM), in which capillary zone electrophoresis was applied, and the average recovery was 100.6%. The relative standard deviation (RSD) of migration time was not more than 2.4%, the RSD of relative migration time was not more than 1.1%, and the RSD of the relative area was not more than 0.51%. Meanwhile, the contrast test has been done by the calibration curve method with an internal standard correlation. The content of morphine in opium powder determined by ACM was the same as that by using the calibration curve method with an internal standard correlated. The study shows that ACM is simple, quick and accurate. PMID- 12541626 TI - [Quantitative determinations of glycyrrhizic acid, glycyrrhetinic acid, morphine and sodium benzoate in compound liquorice tablets by capillary zone electrophoresis]. AB - Capillary zone electrophoresis (CZE) was used to quantitatively determine the contents of glycyrrhizic acid, glycyrrhetinic acid, morphine and sodium benzoate in compound liquorice tablets. The detection wavelength was 228 nm and the applied voltage was 14 kV. Borax solution of 50 mmol/L was used as the background electrolyte and hydrochlorothiazide as the internal standard. There were all good linear relationships between the concentrations and the relative areas of glycyrrhizic acid, glycyrrhetinic acid, morphine and sodium benzoate. Their average recoveries were 98.2%, 97.3%, 97.1% and 97.5%, respectively. The method is simple, rapid and accurate. PMID- 12541627 TI - [Determination of caffeine in teas by supercritical fluid chromatography]. AB - A rapid supercritical fluid chromatographic method for the determination of caffeine in tea is described. It is shown that a mobile phase of carbon dioxide, modified with 5% methanol achieved excellent results. The method is simple in the sample preparation. Other components contained in the sample do not interfere the measurement. It can be used to determine caffeine in tea rapidly. PMID- 12541628 TI - [Establishment of comprehensive fingerprint database of traditional Chinese herbs]. AB - Quality control of traditional Chinese herbs by fingerprint is being paid close attention in these years. A database software has been designed and built to store these fingerprints and other relevant information. The use of this database is very simple and convenient for ordinary analysts who have no experience on professional database software. This software could be used under many kinds of operation systems such as Windows 95, 98, 2000 and Windows NT 4.0, 5.0. By now, nearly 30 fingerprints and their relevant information of ten kinds of traditional Chinese herbs have been input into this database, and the number of fingerprints in this base will be continuously increased in the future. This database is helpful for the analysis and quality control of traditional Chinese herbs. PMID- 12541629 TI - [Determination of aconitine in biological samples]. AB - The determination of aconitine in biological samples by using TLC, HPLC and animal tests is described. TLC is simple, easy to manipulate and highly sensitive. The minimum detection limit was 0.3 microgram, so it can be used as a preliminary screening method. While detecting by HPLC, the characteristic UV absorption spectrum of aconitine and animal tests were used as the qualitative methods. The characteristic UV absorption wavelengths were (228 +/- 2) nm and (275 +/- 2) nm. The linear range of calibration curve was 2 mg/L-50 mg/L. The coefficient of variation was 0.9996. It is suitable for applications in forensic field. PMID- 12541631 TI - [Determination of moth-proof agent residues in textiles with headspace solid phase microextraction and gas chromatography-mass spectrometry]. AB - A procedure is described for the identification and determination of volatile moth-proof pesticide residues, dichlorobenzene and naphthalene, in textile products by headspace solid-phase microextraction (HS-SPME) coupled with GC/MS in selective ion monitoring mode (SIM). Several standard fabrics (ISO 105/F01-1982) are used as sample matrices to reduce the target compound-matrix interaction. Prior to SPME, 0.2 g textile sample is immersed in 3 mL boric acid-citric acid sodium acid phosphate buffer (pH 2.2) solution with 300 g/L sodium chloride and 5% (volume fraction) methanol for supersonic treatment in water bath(50 +/- 1) degrees C. Then an SPME fiber holder is used manually by passing the needle through the septum of the sample vial, depressing the plunger and lowering a fused silica fiber coated with 100 microns polydimethylsiloxane (PDMS) into the headspace right above the sample solution. Adsorption equilibrium is almost established within 5 minutes with magnetic stirring at (40 +/- 1) degrees C. After sample adsorption, draw SPME fiber into the needle, and withdraw the needle from the sample vial and immediately introduce it into the chromatograph injector, where the adsorbed analytes are thermally desorbed for 3 minutes at 230 degrees C and delivered to a capillary GC column for analysis by mass detector. The method provides data of very good linearity with low coefficients of variation for volatile target compounds like p-dichlorobenzene and naphthalene. For all the fabrics tested, typical limits of detection (LOD) were 1 microgram/kg and average recoveries from fortified sample fabrics were between 83.6%-115.2% and the relative standard deviations of the residues were 8.1%-9.8%. PMID- 12541630 TI - [Extraction and gas chromatographic analysis of residual trimethylamine(TMA) in the bakelite-type macromolecule complex material]. AB - A method for the determination of residual TMA in the bakelite-type macromolecule complex material has been established. The whole process includes sample pretreatment, TMA extraction and GC analysis. After the samples have been pretreated, the TMA in them was extracted at (38 +/- 2) degrees C for 16 hours. Then the content of TMA in the liquid can be determined directly. From the experimental results we can conclude that the whole method is cheap, simple and accurate. It meets the requirement of microanalysis and can be used to determine real samples. PMID- 12541632 TI - [Determination of methanol and fusel oils in alcohol beverages using headspace solid-phase microextraction and gas chromatography]. AB - A method for the determination of methanol and fusel oils in alcohol beverages using headspace solid-phase microextraction and gas chromatography (HS-SPME-GC) is presented. The solid phase was a coated epoxy resin. The extraction and chromatography conditions were optimized. Limits of detection were 0.02 mg/L-0.04 mg/L and relative standard deviations were in the range of 1.4%-4.1%. The proposed method showed better sensitivity in comparing with direct headspace gas chromatography(HS-GC, the National Standard method). This method was applied to evaluate real samples. The spiked recoveries in beer, wine and functional alcohol samples ranged from 80.8% to 110.3% for methanol and fusel oils. The results by HS-SPME-GC and HS-GC for alcohol samples coincided very well. The proposed method is simple, fast and accurate with high reproducibility, high sensitivity and low cost. It extends the applications of SPME. PMID- 12541633 TI - [Determination of sodium phenytoin in plasma by high performance capillary zone electrophoresis]. AB - A method to determine sodium phenytoin in plasma by high performance capillary zone electrophoresis (CZE) has been established. CZE was performed in 25 mmol/L phosphate buffer solution (pH 12) at 20 degrees C. The capillary column size was 75 microns i.d. x 30 cm (effective length). Sodium phenytoin was quantified with UV detector at 200 nm. The plasma was extracted by ether after plasma was processed with proteinase K for 3 h at 55 degrees C. Then the extract was evaporated to dryness. The residue was reconstituted in 1 mL of running buffer. The average recovery was higher than 95%. The precisions of intra-day and inter day were 3.1% and 4.7% respectively. The detection limit of sodium phenytoin was 0.6 mg/L. PMID- 12541634 TI - [Determination of the point of gene mutation in two types of tissue from the aminoglycoside antibiotics induced deaf patients]. AB - OBJECTIVE: To elucidate the gene mutation presents in hair follicle cells, just as blood cells, in the aminoglycoside antibiotics induced deaf patients(AAID). METHOD: Mitochondrial DNA of blood cells and/or hair follicle cells from 8 members of 3 aminoglycoside antibiotics induced deafness families were analysed using PCR-SSCP and Alw 26 I digestian. RESULT: Our result showed that a gene mutation at nucleotide 1555 in 12 S rRNA of mitochondrial DNA appeared in blood cells and/or hair follicle cells of 7 subjects among those 8 members. Only a father with normal hearing did't exhibit such a mutation. CONCLUSION: This indicated that hairs could be used as a sample instead of blood to perform gene examination for AAID. PMID- 12541635 TI - [Application of posterior canal wall reconstruction and mastoid obliteration]. AB - OBJECTIVE: To explore the obliteration of the old mastoid cavity and reconstruction of the posterior canal wall after radical mastoidectomy. METHOD: Posterior canal wall reconstruction and mastoid obliteration of 11 ears of 10 patients with old mastoid cavity for 1 to 6 years after radical mastoidectomy were done using the pedicle postauricular composite skin flap (PPCSF), and their tympanoplasty were performed at the same time. Follow-up was performed at 3 to 30 months. RESULT: All PPCSFs survived. The old mastoid cavity of 11 ears vanished and reconstructed posterior canal wall remained in normal position without retraction. The drying ear time took 2 to 3 weeks postoperatively, and the symptoms of the radical cavity disease after radical mastoidectomy were cleared up nearly, and the average hearing threshold decreased 13.6 dB HL. The otorrhea and perforation of tympanic membrane resulted from infection happened to 1 of 11 ears three month after the operation. CONCLUSION: The PPCSF is an effective method in the obliteration of old mastoid cavity and reconstruction of the posterior canal wall. It can reduce drying ear time postoperatively, treat the radical cavity disease after radical mastoidectomy and is beneficial to the tympanoplasty at the posterior external at the same time. PMID- 12541636 TI - [The therapy of children with secretory otitis media by myringotomy with grommet insertion and the relevant management]. AB - OBJECTIVE: To improve the diagnosis and treatment level of secretory otitis media (SOM) of children. METHOD: Clinical information of 80 children with SOM were collected and analysed. All of the 80 cases (144 ears) were treated by myringotomy with Grommet insertion. Among them 30 cases by adenotonsillectomy. 10 cases by palatoplasty. Secretions of 54 ears were carded out by bacterial culture. RESULT: 80 cases were followed up for 6 to 12 months. 120 ears of 144 were cured(83%), with hearing improved. The tympanograms and hearing threshold of pure tone were found to be normal. The Gram's positive and negative germs of bacterial culture were 20.7% (6/29 samples) and 24% (6/25 samples) respectively. CONCLUSION: Children with secretory otitis media should be treated as early as possible to improve the patients' hearing and develope their speech. PMID- 12541637 TI - [Cytokine expression in effusions of secretory otitis media]. AB - OBJECTIVE: To assess the cytokine level in effusions of secretary otitis media and it's relationship with clinical index. To further study the inflammatory mechanism of otitis media with effusion. METHOD: The concentration of TNF-alpha and TNFsolR II was quantitatively determined using enzyme-linked immunosorbent assay and it was correlated with clinical index in otitis media with effusion using stepwise regression analysis. RESULT: TNF-alpha and TNFsolR II was found in 100% effusions. Mean values of TNF-alpha were (3.42 +/- 2.25) ng/g total protein. Mean values of TNFsolR II were (405.80 +/- 216.53) ng/g total protein. Mean values of TNFsolR II index was (154.18 +/- 90.45) U. There was a significant decrease of TNFsolR II and TNFsolR II index in patients who recurred (P < 0.05). CONCLUSION: Cytokine TNF-alpha and TNFsolR II are important inflammatory medium which exist in all otitis media effusions. The decrease of the level of TNFsolR II and TNFsolR II index present significantly in middle ear effusions with recurrence, but there is no any relation of TNF-alpha in middle ear effusions with recurrence. PMID- 12541638 TI - [Study of blood dynamics of ophthalmic artery in patients with sudden sensorineural hearing loss using color Doppler flow imaging]. AB - OBJECTIVE: To investigate the relationship between the pathogenesis of sudden sensorineural hearing loss (SSHL) and the disorder of blood circulation in inner ear. METHOD: Blood dynamics of the ophthalmic artery were studied quantitatively using color doppler imaging in 34 patients with SSHL. RESULT: Compared with 34 self-controls and 15 normal controls, 28 patients (82.4%) with SSHL had significantly lower blood flow velocities and higher resistance indices (P < 0.05), and there was no significant difference between the self-control group and the normal control group (P > 0.05). CONCLUSION: The study suggested that the blood situations--the decreased blood flow velocities and perfusion and increased resistance of ophthalmic artery in patients with SSHL maybe play a role in the pathogenesis of SSHL. PMID- 12541639 TI - [One stage otoplasty treatment for congenital malformations of the auricle and middle ear]. AB - OBJECTIVE: To investigate the application of one stage otoplasty for congenital atresia of the external acoustic canal and malformations of the middle ear and the auricle. METHOD: Patients with the ear malformations were given surgical reconstruction of one stage otoplasty. The auricle was reconstructed with the rib which was encapsulated with the superthin temporal flap. According to the malformations of the middle ear in patients, I style tympanoplasty and III style tympanoplasty were carried out respectively. All patients were performed myringoplasty with temporal fascia and reconstructed the external acoustic canal with full thickness skin-grafting. RESULT: A long term follow-up (4-6 years) demonstrated that 11 ears were survival of which 8 ears figuration were ideal. The hearing improvement was observed in all patients. CONCLUSION: One stage otoplasty is effective for treatment of the congenital malformations of the external and middle ear. PMID- 12541640 TI - [Confirmation of patulous eustachian tube syndrome by Tubo-tymanoaerodynamic graphy]. AB - OBJECTIVE: To compare the advantage of tympanogram, Morimitsu's method and Tubotymanoaerodynamic graphy (TTAG) in the confirmation of patulous eustachian tube syndrome. METHOD: Twenty ears with patulous eustachian tube syndrome diagnosed clinically were selected. The tympanogram, Morimitsu's method and TTAG were examined in these ears and the positive rate was estimated. RESULT: The confirmation of patulous eustachian tube syndrome by tympanogram, Morimitsu's method and TTAG was 5, 12 and 20 ears in 20 ears with patulous eustachian tube syndrome. The positive rate was 25%, 60% and 100% respectively. TTAG also was useful in following-up. CONCLUSION: TTAG is an important method for diagnosis and following-up in patulous eustachian tube syndrome. PMID- 12541641 TI - [Evaluation of the effects of two operative option in the treatment of orbital fractures]. AB - OBJECTIVE: In order to explore the treatment and it's clinical effects of orbital fractures. METHOD: 34 patients were separately treated with nasal endoscopic surgery and external nose incision surgery. RESULT: The total effective rate of nasal endoscopic surgery and external nose incision surgery was 90.0%, 91.6% respectively. There was no significant difference between the two groups (P > 0.05). CONCLUSION: Treating orbital fractures under nasal endoscopic surgery has advantages such as no incision on face, less damage, clear view and easy recheck after operation. It is the best way to treat this kind of disease. PMID- 12541642 TI - [Vowel selection effects on the parameters of acoustic analysis]. AB - OBJECTIVE: To investigate if there is difference about the parameters of acoustic analysis with the different vowel |a|, |i|, |ae| and decide which vowels is best fitted to the acoustic analysis. METHOD: 40 normal and 130 pathologic voice patients accepted acoustic analysis. Each acoustic parameters such as jitter, shimmer, NNE, SDF0, SNR were compared with different vowels. RESULT: Jitter, shimmer, NNE with |i| were much lower than that with |a| and |ae| in normal cases and light hoarseness group. While the acoustic parameters with |i| were much higher than that with |a| and |ae| in moderate and heavy hoarseness group. CONCLUSION: |a| and |ae| were the preferred vowels for normal and light hoarseness group, while the |i| could supply some useful suggestion for the moderate and heavy hoarse patients. PMID- 12541643 TI - [Clinical studies on sternocleidomastoid myoperiosteal flap for cervical tracheal reconstruction]. AB - OBJECTIVE: To introduce the experience of repairing the defect of cervical trachea wall by using the sternocleidomastoid myoperiosteal flap after the anterior or posterior wall of cervical trachea was invaded by cervical neoplasm. METHOD: Between 1989 to 1998 the sternocleidomastoid myoperiosteal flap was applied in 12 patients with different diseases, among which 3 cases were thyroid carcinoma 5 cases were laryngeal carcinoma, 4 cases were cervical esophageal carcinoma. RESULT: The operation was successful. 12 patients were decannuated and had normal exercise tolerance. The time from reconstruction to decannulation was ranging from 20 days to 6 months. CONCLUSION: The sternocleidomastoid myoperiosteal flap is an ideal transplant for cervical tracheal reconstruction. PMID- 12541644 TI - [The curative effect of livostin spray on treating allergic rhinitis of children]. AB - OBJECTIVE: To observe the effect of Livostin spray on children's allergic rhinitis and to search the mechanism of treating allergic rhinitis. METHOD: 113 patients were treated with Livostin spray (Livostin group) or normal saline spray (control group). RESULT: The total efficiency of Livostin group in treating allergic rhinitis is above 95.1% and that of the control group is 25.0%. Initial time of starting effect of Livostin (72.1%) is in 1 minute, and that of the control group (mostly 23.1%) is in 3 minutes. The keeping curative effect time of Livostin spray is mostly (72.1%) above 5 hours and that of the control group is mostly (30.8%) in 3 hours. After 2 weeks, the eosinophilic granulocyte number in nose's secretion of Livostin group is obviously reduced (P < 0.05). CONCLUSION: Livostin is better than control group in relieving symptoms, keeping curative effect and safety, so Livostin is one kind of effective drug in treating children's allergic rhinitis. PMID- 12541645 TI - [The effects of iron supplementation on auditory brain-stem response with iron deficiency anemia in rats]. AB - OBJECTIVE: To evaluate the therapeutic methods of sensorineural hearing loss with iron deficiency anemia and their effects. METHOD: Sixty-eight Wistar rats were randomly divided into 3 groups: Group A (iron deficiency group A) 24 rats, group B (iron deficiency group B) 24 rats and group C (control group) 20 rats. Group A and B were fed with iron deficiency diet for 6 weeks. By analyzing ABR and DPOAE, hearing loss was detected in group A and B. Then, group A was given iron supplementation diet and hyperbaric oxygen therapies, and group B was given iron supplementation diet only. RESULT: The auditory thresholds and period latencies (PL) of I waves of ABR and DPOAE amplitude in 2 kHz and 3 kHz within group A and B were significantly different before and after treatment (P < 0.05). But the effects of treatment between these two groups had not significant differences (P > 0.05). CONCLUSION: It suggested that good results can be obtained by early iron supplementation in iron deficiency hearing loss cases. Hyperbaric Oxygen therapy may not be necessary. PMID- 12541646 TI - [Study on the working noise in BYPC and the effects caused by working noise on the workers' vestibular and auditory function]. AB - OBJECTIVE: In order to observe the kinds and intensity of the working noise of Yansan Petrochemical Co. and the effects caused by the working noise on the workers' vestibular and auditory function. METHOD: The intensity and frequency of the working noise were recorded by exactolnoisemeter in the workshop. One hundred and seventeen workers were tested in routine pure-tone tested method. The SPVN and ABR were tested within fifty-one workers of all. RESULT: The working noise of Yansan Petrochemical Co. belongs to the broad band and steady noise. The intensity of the working noise were during 85.7-104.0 dB(A) and the main frequency were during 1-8 kHz. About 59 percent workers who exposed to the working noise had hearing loss. The most hearing-loss were in the high frequency. The hearing-loss of speech frequency were slight. Workers who have more six years standing have obviously increased hearing-loss than the workers who have less five years standing. There were significant differences the ABR thresholds and wave-interval between the tested and controlled groups. The SPVN and CP were abnormal in more than 17.4 percent workers with hearing loss. CONCLUSION: The working noise of Yansan Petrochemical Co. belongs to the broad band and steady noise. Working noise can lead to workers' hearing loss of certain degrees who exposed in the noise for a long time. Obvious correlation was not defined between the hearing-loss and the abnormal vestibular response group. PMID- 12541647 TI - [The determination of porphyrin carbon isotope composition by gas chromatography isotope ratio monitoring mass spectrometry technique]. AB - The porphyrin carbon isotope composition can be used to explore the precursor of porphyrin, oil-oil and oil-source rock correction and calculation of paleo P CO2. The conventional method is limited because of its time consuming and large sample size (several mg of individual porphyrin) required. Therefore, it hampers the application of porphyrin carbon isotope composition into the chemistry and geoscience. The present paper describes a quantification method to prepare bis (tert-butyldimethylsiloxy) silicon (IV) [(TBDMSO)2Si(IV)] porphyrin which is sufficiently volatile at 300 degrees C and can be used for GC-IRMS analysis. The analysis of carbon isotope composition of aetio I as the form of free base, nickel, demetalization derivative, silicon(IV) and (TBDMSO)2Si(IV) have shown that aetio I porphyrin has no obvious isotope fractionation in the whole synthesis procedure for (TBDMSO)2Si(IV) porphyrin. The carbon isotope study on the porphyrin mixtures of aetio I and OEP indicates that isotope exchange between porphyrins during the synthesis of (TBDMSO)2Si(IV) porphyrin is absent. The method can be applied to the determination of porphyrin carbon isotope compositions. The advantages of the method are time saving, less sample size and lower standard deviation. PMID- 12541648 TI - [Chromatographic behavior and purification of synthetic huwentoxin-I]. AB - A comparative study was made to investigate the chromatographic behavior and purification of chemically synthesized and native polypeptide neurotoxin huwentoxin-I (HWTX-I) by means of reversed-phase HPLC and ion-exchange chromatography. The results showed that the synthetic HWTX-I in crude product has a longer retention time than the native HWTX-I, but has the same retention time as the denatured HWTX-I, suggesting it being in a state of complete denaturation which was confirmed by the MALDI-TOF MS analysis. It has also been discovered that, before complete purification, the synthetic HWTX-I, after oxidative treatment, showed wider and more asymmetric peaks than the native HWTX-I that was denatured and then renatured under the same experimental conditions, which suggests that there is even greater conformational unhomogeneity in the synthetic HWTX-I. It was inferred that, in addition to the wrong disulfide bonds formation during oxidative folding, the racemization during chemical synthesis resulted in more stereoisomers of synthetic HWTX-I. It was needed for the renatured synthetic peptides to be purified using different methods alternatively. PMID- 12541649 TI - [Simultaneous determination of various food additives by high performance liquid chromatography]. AB - A novel method is proposed for the simultaneous separation and determination of eight food additives, acesulfame potassium (AK), aspartame (ASP), benzoic acid (BA), caffeine (CA), saccharin sodium (SA), sorbic acid (SOR), theobromine (TB) and theophylline (TP) by reversed-phase high performance liquid chromatography. The separation was achieved within 23 min by using an Alltech Econosphere C18 column with 10 mmol/L NaH2PO4(pH 4.00)-acetonitrile (90:10, V/V) as mobile phase. The qualification and quantitation were accomplished by using a photodiode array detector. The detection limits (S/N = 3) for all analytes were below mg/L level. Under the experimental conditions, other common food additives and organic acids such as cyclamate, citric acid, malic acid, tartaric acid and ascorbic acid, did not interfere with the determination. The method has been successfully applied to the analysis of various foods as well as pharmaceutical preparation, and the average recoveries for real samples ranged from 78.5% to 107.2%. PMID- 12541650 TI - [Enantioseparation of twelve pharmaceutical racemates with high performance capillary electrophoresis using L-leucine as chiral selector]. AB - A rapid enantiomeric separation method using L-leucine as chiral selector was established. Capillary zone electrophoresis (CZE) has been used for the enantiomeric separation of twelve pharmaceutical racemates with bare fused silica capillary and employing L-leucine as chiral selector. The enantiomeric resolution was influenced by L-leucine concentration and pH of background electrolyte (BGE). The effects of the BGE types and concentrations on the enantiomeric separation were also investigated. The results showed that in the solution containing 50 mmol/L borax and 70 mmol/L L-leucine (pH 9.0), all the twelve drugs were on baseline separated in less than 11 minutes. PMID- 12541651 TI - [Resolution of overlapping chromatographic peaks by radial basis function neural network]. AB - A new algorithm-resolution of overlapping chromatographic peaks by radial basis function neural network(RBFNN) is presented. A two-phase genetic algorithm(GA) which has robustness and random globe optimization is used to train RBFNN so that it has the ability on the resolution of overlapping chromatographic peaks. The two-phase genetic algorithm involves two procedures: training structure and optimizing parameter. The first procedure uses GA to train the architectures of RBFNN, the second procedure uses gradient descent to train the center(tR) and the width(sigma) of RBFNN. The alternate use of these two procedures makes the network having the ability to learn structure, therefore makes itself adaptable to resolution of the chromatographic peaks with unknown number of components. The method proposed here needs no artificial interference, not only has it robustness and globalism, but also the ability of accurate resolution to completely overlapped chromatographic peaks. The simulation experiments show that this method is more accurate than other methods. PMID- 12541652 TI - [Quantitative structure-retention relationship (QSRR) studies of polychlorinated dibenzofurans (PCDFs) on diverse gas chromatographic stationary phases on a set of novel molecular distance edge vector]. AB - By taking phenyl as a pseudo atom, there are three types of atom in the polychlorinated dibenzofuran (PCDF) molecule, being chlorine atom, oxygen atom and pseudo atom. The chemical structure of PCDF congeners is depicted by a novel molecular distance-edge vector(VMDE, mu in short), developed in our laboratory, which consists of the modified VMDE parameters based on the identical group as a pseudo atom instead of a traditional atom. Furthermore quantitative structure retention relationships (QSRR) between the new mu vector and gas chromatographic (GC) retention behavior of PCDFs are generated by multiple linear regression method for various stationary phases. Four models, each of which is constructed by using all sample sets, with high correlation coefficient, r > 0.98, are developed for three columns (DB-5, SE-54, OV-101). In order to test the equation stability and prediction ability of each model, it is essential to perform a cross validation (CV) procedure. Satisfactory CV results have been obtained by using one external predicted sample every time with high correlation coefficients, r > 0.97. These results show that the new mu vector has high structural selectivity and good property relativity, and it is easy to calculate the mu vector. And the QSRR models have high relative coefficients, good stability and good predictability. PMID- 12541653 TI - [Topological research of Kovats indices for amines]. AB - A novel connectivity index(mQ) is defined as mQ = sigma (ti.tj.tk....)-0.5 in this paper. 0Q and 3Q have very good correlation for Kovats indices of 22 amines on three stationary phases(OV-101, OV-225 and NGA). Their linear regression equations are proposed as follows: IOV-101 = 118.341 + 197.854 x 0Q + 448.773 x 3Q, r = 0.9733; IOV-225 = 249.218 + 1,815.760 x 3Q + 343.222 x 1Q, r = 0.9746; INGA = 382.196 + 2,004.277 x 3Q + 318.416 x 1Q, r = 0.9734. These models can better elucidate the change rule of Kovats indices for the amines. Furthermore, a modified Jackknife's test was performed to validate the model robustness. PMID- 12541654 TI - Rapid analysis of momordicoside A in bitter melon by high performance liquid chromatography following solid phase extraction. AB - A rapid, simple and accurate method for the determination of momordicoside A has been established using solid phase extraction (SPE) followed by high performance liquid chromatography (HPLC). Sample was processed by SPE on a Carb cartridge (3 mL/250 mg), and was then determined by HPLC on a C18 column (4.6 mm i.d. x 250 mm, 5 microns) with V(acetonitrile):V(methanol):V(50 mmol/L potassium dihydrogen phosphate buffer) = 25:20:60 as mobile phase (0.8 mL/min) and UV detection at 208 nm. The calibration curves were linear from 10 mg/L to 1,000 mg/L (r2 = 0.9992). The analytical method was shown to be highly reproducible, giving all of the relative standard deviations and relative mean errors less than 10% for both intra-day and inter-day determinations. The absolute recoveries were greater than 90%. PMID- 12541655 TI - [Technique and advance of comprehensive two-dimensional gas chromatography]. AB - Many analytical problems require more resolution than the conventional single column chromatographic technique can provide. In such cases the separation power can be enhanced by using more than one separation technique or mechanism. The sample is then dispersed in different time dimensions. The resolution obtained depends strongly on the difference between these dimensions. The highest resolution is gained when there is no correlation between the separations, the dimensions being orthogonal to each other. Comprehensive two-dimensional gas chromatography (GC x GC) provides a true orthogonal separation system in which a modulator serially couples two columns containing dissimilar stationary phases. It focuses and subsequently reinjects components eluting from the first column into the second one. The system generates a peak capacity that is approximately equal to the product of the peak capacities of the two individual separation systems. In this paper, technique and instrumental considerations of GC x GC are discussed. The three designs of contemporary GC x GC systems are presented and compared. A number of typical applications on complex samples such as petroleum products and environmental pollutants are also cited. Finally, the future perspectives of GC x GC are simply discussed. PMID- 12541656 TI - [A study on the contents of fat and fatty acids in five shellfishes]. AB - The contents of fat and fatty acids in Callista erycina (Linnaeus), Paphia (Paratapes) undulata (Born), Meretrix meretrix (Linnaeus), Chlamys farreri (Jones et Preston) and Patinopecten yessoensis (Jay) were studied. Fat was extracted with Bligh & Dyer (B&D) method. The lipid classes were transesterified with potassium hydroxide in methanol. Fatty acid methyl esters (FAMEs) were assayed with GC-MS and polar capillary column (HP-INNOWax 30 m x 0.25 mm i.d. x 0.25 micron). GC injector temperature was 220 degrees C. The column temperature was programmed from 150 degrees C (1 min) to 200 degrees C at 10 degrees C/min and then from 200 degrees C to 250 degrees C at 2 degrees C/min. FAMEs were identified by MS library, and part by their standards. Total identified fatty acids were over 99% for all samples. Fat contents of them were all over 1% by wet samples. And ratios between omega-3PUFA and omega-6PUFA were above 2 by and large. Patinopecten yessoensis (Jay) contains more fat and the valuable fatty acids, EPA and DHA. It is suitable to use it as the source of EPA and DHA. PMID- 12541657 TI - [Study on gas chromatographic method for the evaluation of residual solvents by using wide bore open tubular columns]. AB - A GC method using wide bore open tubular columns was developed to evaluate residual organic solvents. Sixteen organic solvents commonly used for pharmaceutical synthesis were analysed. The effects of different polarity columns and extraction solvents on the assay of residual solvents were studied in this work. It showed that most solvents can be well separated from each other on non polar or less polar wide-bore columns. Extraction solvents differed in their extraction recovery and peak shape. Satisfactory results can be obtained when the proposed method was employed and a suitable extraction solvent was selected. The procedures summarized in this paper can be used as a guidance for the evaluation of residual organic solvents in bulk pharmaceuticals. PMID- 12541658 TI - [Study on analysis of trace phenol in small amount of aqueous sample by liquid liquid micro-extraction and gas chromatography]. AB - A method for the liquid-liquid micro-extraction and gas chromatographic analysis of trace phenol in small amount of aqueous sample has been developed. The influences of acid and salts on recovery rate of phenol were examined. And, the influences of injection speeds and the conditions of silica wool in injection port on the quantitation precision were tested. With a direct injection port, an FID and a DB-1 capillary column of wide bore, a detection limit of 1 microgram.L 1 can be obtained employing 8 mL water sample, 160 microL ethyl acetate and 3.5 g ammonium sulfate. The added recoveries were in the range of 95.0%-98.5%. The relative standard deviations were in the range of 2.8%-3.3%. This method is simple, convenient, rapid, accurate and practical. PMID- 12541659 TI - [Determination of fatty acids in Malania oleifera oil by gas chromatography-mass spectrometry]. AB - Fatty acids in the Malania oleifera oil of Yunnan were esterified by sulfuric acid-methanol and analyzed by GC-MS. Seventeen components were separated and seven acids as methylesters were identified and determined. They were hexadecanoic acid (1.50%), 9c-octadecenoic acid (21.54%), 11c-eicosenoic acid (7.04%), 13c-docosenoic acid (18.29%), docosanoic acid (1.33%), 15c-tetracosenoic acid (40.92%), tetracosanoic acid (2.14%). There were 7.24% components unidentified. PMID- 12541660 TI - [Analysis of group composit of crude oil by on-line coupling of capillary HPLC HTGC]. AB - The group composition of crude oil was analyzed by on-line coupled capillary HPLC HTGC. After removal of asphaltenes, the oil sample was separated by capillary HPLC into aliphatics, aromatics and resins. The interface cut and stored each fraction, and transfered them into GC sequentially. The group contents of oil were determined by FID. The error of reproducibility(RSD) was less than 3%. The method is accurate, time saving, and easy for operation. It is very important for quality control and development of new and better products in crude oil. PMID- 12541661 TI - [Preparation of restricted-access media--alkyl-diol silica (ADS) by an improved method]. AB - Alkyl-diol silica (ADS) is a kind of restricted-access media, which can be used for direct injection and analysis of complex biological samples containing bio macromolecules. A new economical procedure has been developed to prepare ADS packing. At first gamma-glycidoxypropyl group was coupled onto the surface of microporous silica gel (Develosil 60-5 mu) to form epoxy-silica. Prepared epoxy silica can react with stearic acid in organic solvent to prepare C18 ester-bonded reversed-phase packing. The packing was packed into a column and then the solution of pancreatic lipase was pumped into the column to create enzymoloysis reaction. The stearyl groups on the surface of packing can be removed by the enzymolysis to form a hydrophilic surface. At the same time, inner surface of micropore remains hydrophobic nature due to size exclusion effect of micropore to enzyme molecules. Chromatographic evaluations were carried out and the typical ADS behavior was confirmed. PMID- 12541662 TI - [Preparation, characterization and evaluation of cellulose tris (3,5 dimethylphencarbamate) chiral stationary phase]. AB - With the deposition of cellulose tris(3,5-dimethylphenylcarbamate) (CDMPC) on aminopropylated silica gel (mean particle size, 5 microns; pore size, 13 nm; surface area, 110 m2/g) by using two different methods (evaporation and precipitation), two chiral stationary phases (CSP1 and CSP2) characterized by elemental analysis and scanning electron micrography were obtained. They were also evaluated by using seven racemic compounds with n-hexane/ethanol(95/5, V/V) and n-hexane/2-propanol (90/10, V/V) as mobile phases. The results showed that the chiral stationary phase CSP1 obtained by the evaporation method had better efficiency and chiral resolution ability than CSP2 by the precipitation method. PMID- 12541663 TI - [Recognition and quantitative contrast characteristic components for root of Chinese angelica]. AB - A method of multi-component quantitative analysis in root of Chinese Angelica has been developed. The analytical mode was reversed-phase HPLC with photodiode array detector. Based on comparison of UV spectra, a software of UV spectra database to assist recognition has been established. The solvent system, a gradient program of mobile phase with 2-propanol/acetonitrile/water can separate a lot of liposoluble components in root of Chinese Angelica. UV detection wavelength was set at 202 nm and the flow rate was 1 mL/min on a Hypersil ODS column. Twenty three components in two roots of Chinese Angelica were recognized using characteristic parameters of chromatography and UV spectroscopy. It is the base of quantitative contrast to recognize many characteristic components without standard components in root of Chinese Angelica. For two samples of root of Chinese Angelica both have 23 characteristic components and the concentrations of some components were similar and those of others were different. The method is convenient and reproducible. It can be used for the quality control of the root of Chinese Angelica without the use of standards. PMID- 12541665 TI - [Study on analysis of concentration of surfactant PS in injected-produced liquor used in ASP flooding with HPLC]. AB - Ion exchanger and ODS bonded silica gel for HPLC packing were prepared. An HPLC method for the analysis of surfactant PS concentration in injected-produced liquor has been established to meet the need of ASP flooding developed in Gudao West Block of Shengli Oilfield, with a minimum detectable limit of 0.4 mg/L, a linear range of 50 mg/L-1,000 mg/L and recoveries of 95.7%-99.8%. This method has provided great technical support in a variety of fields to the design of the prescription for ASP flooding, the regulation of injection measurement, the quality control of products and the study on the mechanism of oil flooding etc. PMID- 12541664 TI - [Separation and determination of ephedrine alkaloids and 2,3,5,6-tetramethyl pyrazine in Ephedra herba by HPLC]. AB - A sensitive and reliable high performance liquid chromatographic method(HPLC) has been developed for the first time for the simultaneous determination of the active ingredients of ephedrine alkaloids and 2,3,5,6-tetramethyl pyrazine (TMP) in Ephedra herba crude drug and two Chinese traditional medicines (Xiao-er qingfeiwan and Lu-si kewan). The HPLC assay was performed on a reversed phase C18 column (Nova-Pak C18, 3.9 mm i.d. x 150 mm) by using methanol-0.02 mol/L KH2PO4 acetic acid-triethyl amine (4:96:0.2:0.01, V/V) as mobile phase for the ephedrine alkaloids analysis and methanol-H2O-acetic acid (35:65:0.5, V/V) as mobile phase for TMP analysis. Regression equations revealed the linear relationships (correlation coefficients: 0.991-0.998) between the peak area of each constituent (E, PE, NE, NPE, TMP) and its concentration. The detection limits for E, PE, NE, NPE and TMP were 0.4 mg/L, 0.1 mg/L, 0.03 mg/L, 0.02 mg/L and 0.03 mg/L, respectively, and the recoveries ranged between 92%-103%. The contents of E, PE, NE, NPE, TMP in Ephedra herba, traditional medicine Xiao-er qingfeiwan and Lu-si kewan were determined respectively. The relative standard deviations (RSD) of the contents ranged between 1.1%-3%. PMID- 12541666 TI - [The analysis of rice total RNA by capillary electrophoresis]. AB - When the RNase was eliminated through backing at 200 degrees C or treated by diethylpyrocarbonate (DEPC), the total RNA of rice could be separated within 15 min using 1.0%T, 0%C linear polyacrylamide as sieving matrix and 7 mol/L urea as denaturant. The tRNA of rice can be separated into two classes and about nine peaks when high concentrated polyacrylamide sieving matrix (5.0%T, 0%C) was used. This technique could provide one of the rapid and accurate methods for the determination of RNA in plants. PMID- 12541667 TI - [Determination of human PDGF-B promoter binding nuclear protein by capillary electrophoresis]. AB - With the use of 1.0% T, 0% C linear polyacrylamide as sieving matrix, 0.25 x TBE(Tris 89 mmol/L, boric acid 89 mmol/L, EDTA 2 mmol/L) as running buffer and 15 degrees C as column temperature, the human PDGF-B promoter binding nuclear protein can be determined within 50 min with good resolution. The results proved that there are two proteins having strong ability binding human PDGF-B promoter, which similar to that in slab gel electrophoresis. This technique can provide one of the rapid and accurate separation methods in the studying of the formation and repression behavior of DNA binding protein based on PDGF gene as the target. PMID- 12541668 TI - [Sensitive and rapid analysis of nitrophenols and herbicides by capillary zone electrophoresis (CZE)]. AB - 11 Nitrophenols and some acidic herbicides have been separated by modified CZE. Good separation of such compounds can be achieved in less than 10 min by simply modifying the run buffer with organic solvents such as methanol, acetonitrile in a concentration of 40% (V/V). The limits of detection (LODs) for nitrophenols by UV-absorption at 215 nm were 0.5 mg/L-1.1 mg/L. Solid phase extraction was used to preconcentrate nitrophenols and some herbicides to reach an LOD at microgram/L levels. Typical samples have been analyzed. PMID- 12541669 TI - [Study on sampling preconcentration method combining electrostacking with isotachophoresis in capillary electrophoresis]. AB - A sampling preconcentration method combining electrostacking with isotachophoresis is presented in this paper. The optimum conditions for electrostacking and isotachophoresis were investigated. With the preconcentration conditions of 70 s electrostacking and 40 s isotachophoresis, two medicines, propranolol and metoprolol, with similar structures were separated by capillary zone electrophoresis. The working voltage for all the steps was 15 kV. Being adjusted to pH 4.0, 30 mmol/L NaAc-HAc, 30 mmol/L beta-alanine-HAc and 1.5 mmol/L NaAc-HAc were employed as the background, viz. leading, terminating and sample buffer solutions respectively. In comparing with the conventional electro migration injection, the enhancement factors in peak height of the present method were about 250 and 160 for propranolol and metoprolol respectively. Total analysis time was similar to that of the conventional one. PMID- 12541670 TI - [Supercritical fluid extraction of beta-elemene under lower pressure]. AB - A method has been developed for the extraction and analysis of the active component beta-elemene from Rhizoma Atractylodis Macrocephalae by supercritical fluid extraction with CO2 containing ethanol. The article discusses some factors such as pressure, temperature and modifier that affect the selectivity. Class selective extraction has been performed by varying the pressure. beta-Elemene could almost be extracted completely at lower pressure (12.0 MPa) and its relative amount was raised from 4.8% under higher pressure to 8.0% for lower pressure. PMID- 12541671 TI - [Determination of alkali metals, ammonium ion and alkyl amines by non-suppressed ion chromatography]. AB - The determination of Li+, Na+, NH4+, K+, CH3NH2, CH3CH2NH2 and CH3CH2CH2NH2 by non-suppressed ion chromatography using sulfamic acid as mobile phase is described. A cation-exchange column was employed for separation with conductivity detection. The effect of the eluent concentration on retention value was studied. The retention value of ions decreased with increasing the eluent concentration. The suitable eluent concentration was 4.0 mmol/L. When 4.0 mmol/L sulfamic acid was used as mobile phase, the detection limits (mg/L) for Li+, Na+, NH4+, K+, CH3NH2, CH3CH2NH2 and CH3CH2CH2NH2 were 0.008, 0.029, 0.027, 0.099, 0.069, 0.163 and 0.417 and the linear ranges of calibration curve(mg/L) were 0.008-20, 0.029 20, 0.027-20, 0.099-30, 0.069-30, 0.163-40 and 0.417-40, respectively. The method has been applied to the determination of tap water samples. The recoveries of added standard were 94.0%-97.6%. PMID- 12541672 TI - [Resolution of overlapping mass spectra of azobiphenyl dyes by subwindow factor analysis technique]. AB - Subwindow factor analysis(SWFA) has been proven to be an effective data processing technique. In this paper, it is applied to the resolution of overlapping GC-MS peaks. A two-component sample of azobiphenyl dyes (3,3' dichlorobenzidine, 4,4'-methylene-bis-(2-chloroaniline)) was prepared and a data set from a mixture of azobiphenyl dyes recorded by GC-MS was obtained. Then the overlapping GC-MS data were resolved by SWFA. The results show that both mass spectrum and chromatogram of components can be resolved from multicomponent overlapping data. The resolved spectra were in good consistence with the mass spectra of standard samples. At the same time, overlapping GC-MS data were also resolved by WFA. In Comparing with WFA technique, SWFA is more convenient, and the speed of resolution could be increased. PMID- 12541673 TI - [Separation of standard proteins and actual samples using cross-axis counter current chromatography]. AB - A system of m (12.5% PEG8000): m (25% K2HPO4) = 1:1 was used to separate two kinds of standard proteins, glycoprotein in Morchella esculenta (L.) and glycoprotein in Lycium barbarum (L.), the upper phase was used as stationary phase, and the lower phase was used as mobile phase. The revolution speed was 500 r/min and the flow rate was 60 mL/h. In comparing with high performance liquid chromatography, cross-axis counter current chromatography can increase load capacity without loss of resolution. The advantage of this method was verified, indicating that cross-axis counter current chromatography was a useful method for biopolymer separation. PMID- 12541674 TI - [Determination of salicylic acid and its hydroxylated products using high performance liquid chromatography and fluorescence detection]. AB - High performance liquid chromatography (HPLC) with fluorescence detector was used to determine salicylic acid (SA) and its main hydroxylated products, 2,3 dihydroxybenzoic acid (2,3-DHBA) and 2,5-dihydroxybenzoic acid (2,5-DHBA). Mobile phase was 30 mmol/L acetate(pH 4.9) and the flow rate was 1.0 mL/min. Low detection limits, 10 nmol/L for 2,3-DHBA, 0.6 nmol/L for 2,5-DHBA and 1.0 nmol/L for SA were obtained. The photolysis system of salicylic acid and H2O2 was investigated. The products in the system were measured and the yields were 45% and 51% for 2,3-DHBA and 2,5-DHBA, respectively. PMID- 12541675 TI - [Effects of blood transfusion on cellular immuno-function in patients with laryngeal carcinoma]. AB - OBJECTIVE: To explore the influence of blood transfusion on cellular immunofunction in patient with laryngeal carcinoma. METHOD: EPICS-XL flow cytometry was used to measure T cell subgroup, NK cell and CD28 in 36 patients with laryngeal carcinoma pre-operation and 2 weeks post-operation. Patients were divided into allotransfusion group and non-transfusion group. Comparison was conducted between the 2 groups. RESULT: 1. Comparison with normal population, decreasing of CD3, CD4, NK cell and CD28 in the 36 patients pre-operation was statistically significant (P < 0.01). 2. Decreasing of CD3, CD4, NK cell and CD28 was statistically significant post-operation (P < 0.05). 3. In the non transfusion group change of CD3, CD4, CD8, NK cell and CD28 post-operation was statistically insignificant (P > 0.05). CONCLUSION: While cellular immunofunction is generally low in patients with laryngeal carcinoma, allotransfusion will reduce further. It makes contribution to spreading and metastasis of carcinoma easier. PMID- 12541676 TI - [Study of pharyngo-cutaneous fistula after total laryngectomy]. AB - OBJECTIVE: To explore the factors which lead to pharyngo-cutaneous fistula after total laryngectomy. METHOD: A retrospective study of 125 patients who underwent total laryngectomy during a 16-year period was carried out. RESULT: The rate of post-laryngectomy pharyngo-cutaneous fistula is 16.0% (20/125), it appears reduced tendency. With single-factor analysis, the factors effect on the rate of post-laryngectomy pharyngo-cutaneous fistula include clinical tumor stage, tumor types, pre-operative radiotherapy dose, intraoperative blood transfusion, operative duration. With all variable logistic model analysis, the factors effect on the rate of pharyngo-cutaneous fistula are clinical tumor stage and pre operative radiotherapy dose, but pre-operative radiotherapy is very possible to be a risk factor (P = 0.0566). With logistic model stepwise regression analysis, the factors effect on the rate of pharyngo-cutaneous fistula are clinical tumor stage and operative duration. CONCLUSION: Pharyngo-cutaneous fistula appears easily in patients with T3, T4 tumor, and have pre-operative radiotherapy, especially have large dose preradiotherapy. For these high risk patients, the duration of operation should be as short as possible to reduce the rate of post laryngectomy pharyngo-cutaneous fistula. PMID- 12541677 TI - [Detection of S-100 protein in laryngeal carcinoma]. AB - OBJECTIVE: To study the relationship between S-100 protein and prognosis of laryngeal carcinoma. METHOD: S-100 protein was observed by immunohistochemical methods using anti-S-100 protein antibody in 68 laryngeal carcinoma and 21 non cancer laryngeal tissues. RESULT: The result showed that S-100 protein positive were found 43 (63.24%) in 68 laryngeal carcinoma. S-100 protein positive cells were interspersed among the tumor cells and in the stroma. S-100 protein were rare in non-cancer laryngeal tissues. The five-year survival rate in patients with S-100 protein positive was significantly higher than that in patients with S 100 protein negative. CONCLUSION: Our result indicated that the detection of S 100 protein is of value for the prognostic analysis in patients with laryngeal carcinoma. PMID- 12541678 TI - [Study on the relation between the cell cycle regulators and laryngeal carcinogenesis]. AB - OBJECTIVE: To investigate the clinicopathological significance of the Cyclin E, p21WAF1/CIP1 and p53 expression in laryngeal carcinogenesis. METHOD: The expression of Cyclin E, p21WAF1/CIP1 and p53 was examined by using immunohistochemical method in normal epithelia (20), dysplastic epithelia (40) and laryngeal cancers (60). RESULT: 1. In normal epithelium, dysplastic squamous epithelium and in squamous cell carcinoma of the larynx, the positive immunostaining rate of Cyclin E protein was 5.0%(1/20), 20.0%(8/40) and 45.0%(27/60) respectively (P < 0.001), and p21WAF1/CIP1 protein was 95.0%(19/20), 75.0%(30/40) and 63.3%(38/60) respectively (P < 0.01), while the rate of p53 was 0, 30.0%(12/40) and 61.7%(37/60)(P < 0.001) respectively. 2. p21WAF1/CIP1 positive expression in highly, moderately and poorly differentiated carcinomas were 76.2%(16/21), 66.5%(19/29) and 30.0%(3/10) respectively (P < 0.05), p21WAF1/CIP1 protein expression was significantly associated with tumour cell differentiation. 3. The positive expression of Cyclin E closely related with the positive expression of p53 (P < 0.05). No correlation was found between p21WAF1/CIP1 expression and p53 expression. CONCLUSION: 1. Cyclin E and p53 could be an early molecular event in the carcinogenesis of larynx. 2. p21WAF1/CIP1 expression is associated with tumour cell differentiation. 3. The mutual mechanism of cell cycle regulators (Cyclin E, p21WAF1/CIP1 and p53) play an important role in the laryngeal carcinoma. PMID- 12541679 TI - [Measurement of microvessel density in laryngeal carcinoma and its clinical significance]. AB - OBJECTIVE: To determine the microvessel density (MVD) in laryngeal carcinoma and its clinical significance. METHOD: Thirty-eight tumor specimens were selected from laryngeal cancer patients from January, 1994 to March, 1996. Histological sections of the tumors were stained immunohistochemically for factor VIII. Using light microscopy, we counted microvessels per 400 x field in the most active areas of tumor angiogenesis. RESULT: 1. The tumor blood vessels, composed of only one layer of endothelium were mainly distributed heterogeneously in the interstitial tissue of laryngeal carcinoma with irregular lumen, poorly developed structure. 2. The MVD in the cancer tissues were statistically higher than that in peritumoral tissues (P < 0.01). 3. The MVD in the cancer tissues in group of patients with metastasis to cervical lymphonodes were statistically higher than in group without metastasis (P < 0.01), the MVD in the cancer tissues in group of advanced cases (III, IV stages) were statistically higher than that in group of early cases (I, II stages, P < 0.01). 4. There was no statistically difference in MVD in the cancer tissue between supraglottic and glottic laryngeal carcinoma patients (P > 0.05). 5. There was no statistically difference in MVD in the cancer tissue among the G1, G2 and G3 group (P > 0.05). CONCLUSION: The laryngeal cancer blood vessels have some characteristics that don't appear in normal vessels. It is suggested that tumor angiogenesis can promote tumor growth and metastasis and MVD may be a new prognostic indicator of laryngeal carcinoma. PMID- 12541680 TI - [Application of voice acoustic analysis in microlaryngoscopic surgery]. AB - OBJECTIVE: To evaluate the relationship between the effect of microlaryngoscopic surgery of vocal polyp and the change of all-parameters. METHOD: Acoustic parameters of 80 patients with vocal polyp were measured by Dr. Speech Sciente for Windows, and analyzed preoperatively and postoperatively. RESULT: The jitter, shimmer and NNE were different significantly between preoperative and postoperative, the NNE and shimmer were more sensitive than jitter. CONCLUSION: The NNE and shimmer of acoustic parameter are objective parameter to evaluate the effect of the microlaryngoscopic surgery of vocal polyp. PMID- 12541681 TI - [Application of video laryngoscope in diagnosis and treatment of laryngeal diseases]. AB - OBJECTIVE: To investigate the value of video laryngoscope in diagnosis of laryngeal diseases. To study the effect of video laryngoscope in treatment of laryngeal diseases. METHOD: Pentax VNL-1530T and Olympus BF-240T video laryngoscope were used in diagnosis of 3628 cases of laryngeal diseases and in treatment of 1221 cases of laryngeal diseases. RESULT: 3628 cases were examined by video laryngoscope, and correct diagnosis was made. 768 cases of vocal cord polyp and 249 cases of vocal cord node were excised, 113 cases of foreign bodies in hypopharynx or larynx were removed. 91 cases of other laryngeal diseases were treated under video laryngoscope. CONCLUSION: Video laryngoscope is a new excellent instrument in diagnosis and treatment of laryngeal disease. PMID- 12541682 TI - [Herpesviridae and laryngeal neoplasia]. AB - OBJECTIVE: To investigate the relationship between herpesviridae and malignant or benign laryngeal diseases. METHOD: 128 paraffin-embedded laryngeal squamous cell carcinoma and laryngeal epithelium hyperplastic lesions were detected by polymerase chain reaction (PCR) and PCR-ISH for herpesviridae. RESULT: HSV-1 was detected in 10 cases by PCR, among them 3 were laryngeal squamous cell carcinoma (LSCC), 1 was carcinoma in situ(CIS), 4 were laryngeal polyps and 2 were laryngeal keratosis. Except 1 LSCC and 1 CIS 8 of 10 cases were positive while detected by PCR-ISH. In benign diseases, signals were shown from basal layer to superficial cell; in malignant lesions, the signals were scattered in the diseases. CONCLUSION: Most of laryngeal diseases were not related to herpesviridae, but HSV-1 may acts as initiator in the development of a few cases. PMID- 12541683 TI - [A report of twice outbreak epidemic of acute tonsillitis and acute pharyngitis]. AB - OBJECTIVE: To discuss the pathogenic factors, prevention and treatment of fulminant epidemic of acute tonsillitis and acute pharyngitis. METHOD: We investigated and analysed the cases of twice fulminant epidemic of acute tonsillitis and acute pharyngitis. RESULT: In the first time, 91 patients among 518 persons were attacted, the sick rate was 17.6%. In the second time, 39 among 295 persons were attacted, the sick rate was 13.2%. The main symptoms were pharyngodynia and fever. Bacterial culture of throat swab of 20 cases of the first time and 17 cases of the second time were carried out. The positive rate of beta-hemolytic streptococcus was 82.5% and 85.3% respectively. CONCLUSION: Environment and individual health condition are important factors in the fulminant epidemic of acute tonsillitis and acute pharyngitis. PMID- 12541684 TI - [Effects of selective attention and contralateral acoustic stimulation on latency of distortion product otoacoustic emissions]. AB - OBJECTIVE: To investigate the effects of selective attention and contralateral acoustic stimulation on latency of distortion product otoacoustic emissions. METHOD: Latency of DPOAE was recorded in 30 ears of 15 normal subjects with and without visual attention task, contralateral acoustic stimulation and combination of both forms of stimulus. RESULT: No significant change was observed in three form of stimulus. CONCLUSION: The effects of selective attention and contralateral acoustic stimulation on latency of DPOAE have to be further studied. PMID- 12541685 TI - [Study of a mutation in connexin 26 gene associated with congenital sensorineural deafness]. AB - OBJECTIVE: So far, at least 39 deafness gene loci have been mapped in human chromosome including the connexin 26 gene coding for a gap-junction protein. This gene is thought to be linked to hereditary non-syndromic sensorineural hearing loss. About 80% of cases of DFNB1 hereditary deafness carry a 30 mer G mutation of connexin 26. METHOD: To investigate this relationship, we obtained DNA samples from 15 cases with autosomal recessive and autosomal dominant forms of non syndromic deafness. In addition, DNA samples were obtained from 252 unrelated subjects with sporadic hearing loss; parents of these subjects were symptom free. We analyzed the coding region for the connexin 26 gene for mutation using PCR SSCP and sequence analysis and PCR-mediated site-directed mutagensis. RESULT: We detected 46 mutations with SSCP. The PCR products that of 5 cases mutations had similar abnormalities in their electrophoresis bands were sequenced. Results from 2 cases of families with hereditary hearing loss and 3 cases of sporadic hearing loss had a G-to-A transversion at nucleotide 79. In addition, 2 cases sporadic hearing loss with 251 delT and 233 delC were found, 35 delG was not detected in 46 cases abnormal PCR products using PSDM assay. CONCLUSION: High mutation rates were found in China deafness population of connexin 26 gene, but the mutation loci is different from those previously reported. Finding hot spot of connexin 26 gene mutation in China population is important for deafness etiologic diagnosis and affect genetic counseling. PMID- 12541686 TI - [Two terms tympanoplasty in chronic suppurative otitis media]. AB - OBJECTIVE: To explore the clinical manifestations and therapeutic effects of treating chronic suppurative otitis media with two terms tympanoplasty. METHOD: 36 patients were retrospectively analyzed. The criterions of success are air room in middle ear, intact eardrum and decrease of A-B gape in audiometry. RESULT: 32 cases of all patients got successful effects, 4 cases failed. The main cause of failure is to fail forming air room in middle ear. CONCLUSION: The two term's tympanoplasty can save the residual hearing in most patient. But it is difficult to get successful result if more mucous membrane in the middle ear disappeared. PMID- 12541687 TI - [Gentamicin for the transtympanic treatment of Meniere's disease]. AB - OBJECTIVE: This study described the method and efficacy of intratympanic gentamicin injection for obstinated, incapacitated Meniere's disease. METHOD: It is a prospective study. Patients selected had classic unilateral Meniere's disease. All had preinjection audiometric and electronystagmography. Injection was accepted every two or three days. It is important to direct the therapy by observing the symptoms closely and checking the audiometric and electronystagmography whenever necessary. RESULT: All the 12 cases undergone the treatment were followed up from 2 to 9 years. According to the criterion of Chinese Medical Association of ENT, the vertigo of Meniere's disease was evaluated. A (completed control): 10 cases; B (largely control): 2 cases. 4 cases said their tinnitus disappeared and 3 cases felt it improved. Hearing loss and vibration illusion vision are two complications. CONCLUSION: The methods is simple, useful and safe, the effect of which is similar to vestibular neurectomy. It is the first choice therapy suitable to Meniere's disease patients, which symptoms keep unchanged after medical treatment or failed in surgery. PMID- 12541688 TI - [Influence of the hearing loss level and auditory sensation curve type on click evoked SP/AP amplitude ratio in Meniere's disease]. AB - OBJECTIVE: To assess the influence of hearing loss level and auditory sensation curve type on click evoked SP/AP amplitude ratio (negative summating potential/action potential) in the diagnosis of Meniere's disease. METHOD: AP and SP was recorded with tympanic ECochG in normal controls (50 ears), patients with sensorineural hearing loss (SNHL) and patients with Meniere's disease (90 ears) which were further divided based upon the degree of hearing loss and audiometric configuration. SP/AP amplitude ratio was calculated and analyzed. RESULT: AP latency in the three groups had no significant differences. AP amplitude in normal hearing subjects was higher than other two patients groups (P < 0.05). Amplitude of SP of control group was higher than that of SNHL group (P < 0.01), but lower than Meniere's disease group (P < 0.01). SP/AP amplitude ratio in Meniere's disease ears was (48.09 +/- 14.83)%, which was significantly higher than that of SNHL group (20.20 +/- 15.00)% (P < 0.01) and normal group (23.85 +/- 8.05)% (P < 0.01). In Meniere's disease ears, SP/AP amplitude ratio was positively correlated to pure tone thresholds (r = 0.5601 P < 0.01). An elevated SP/AP ratio was found in 73% of the Meniere's ears and 6% of the SNHL ears. SP/AP ratio among the different levels of hearing loss in Meniere's disease ears, of the ears with mild hearing loss was lower than those with moderate (P < 0.01) and severe hearing loss (P < 0.01), but between later groups there was no statistical significant difference (P > 0.05). Positive rate of SP/AP ratio was 63% in mild, 83% in moderate, 73% in severe hearing loss ears respectively. SP/AP amplitude ratio in patients with low tone auditory curve type was lower than that with flat type(P < 0.05), the latter had no significant difference (P > 0.05) compared with high tone type. Positive rate was 67.57% in low tone type, 75% in high tone type and 78% in flat tone type respectively. CONCLUSION: Results suggests the increase of SP/AP ratio was of clinical significance in the diagnosis of Meniere's disease, and was with advancing pure tone thresholds in certain range of hearing loss levels. Click evoked SP/AP amplitude ratio was affected by degree of hearing loss and auditory sensation curve as well, perhaps the influence of the former on SP/AP ratio was considerable significant. PMID- 12541689 TI - [A study on the state of cell proliferation and apoptosis in human middle ear cholesteatoma]. AB - OBJECTIVE: To study the state of proliferation and apoptotic cell death in cholesteatoma. METHOD: 20 cholesteatoma samples and 10 normal external ear canal skin samples were studied with immunohistochemical SABC technique and TUNEL technique. RESULT: Immunohistochemical analysis using a monoclonal antibody to proliferation cell nuclear antigen (PCNA) demonstrated the presence of proliferating cell in the basal cell layer of normal external ear canal skin, whereas in the cholesteatoma tissue samples larger numbers of proliferation cell nuclear antigen-positive cells were also observed in the prickle and granular cell layers. The PCNA labeling index and the mean optical density were (10.52 +/- 2.65)% and (0.1340 +/- 0.0363) in normal external ear canal skin samples and (36.91 +/- 22.77)% and (0.2427 +/- 0.0586) in cholesteatoma tissue samples. In cholesteatoma tissue samples and normal external ear canal skin samples, apoptotic cell were observed in the prickle and granular cell layers. The apoptotic indexers were (27.50 +/- 12.50)% in cholesteatoma tissue samples and (9.96 +/- 3.86)% in skin samples. CONCLUSION: The results demonstrate the rates of apoptotic cell death and cell proliferation in cholesteatoma are higher than those in normal skin. Increased cell proliferation and cell death will contribute to increased keratous debris. PMID- 12541690 TI - [Superoxide dismutase and cytokines in otitis media with effusion]. AB - OBJECTIVE: To study the role of superoxide dismutase (SOD) and cytokines in the pathogenesis of secretory otitis media(SOM). METHOD: The content of SOD, interleukin-6(IL-6), interleukin-8(IL-8) and tumor necrosis factor alpha (TNF alpha) in blood plasma and middle ear effusion (MEE) were measured in 64 (90 ears) patients respectively. RESULT: SOD, IL-6, IL-8 and TNF-alpha were found in 88.9%, 86.8%, 81.5% and 74.6% of MEI respectively. SOD, IL-6, IL-8 and TNF-alpha of blood plasma in SOM group were higher than those in normal control group (P < 0.05). The concentration of SOD, IL-6, IL-8 and TNF-alpha MEE was remarkably higher than that in blood plasma (P < 0.01). It revealed that the shorter the course, the higher the concentration of IL-6 or IL-8 in MEE (P < 0.01, or P < 0.05), and the longer the course, the higher the concentration of SOD or TNF alpha in MEE (P < 0.01, P < 0.05). IL-6 and IL-8 concentration of MEE in serous fluid group was remarkably higher than those in the mucous fluid group (P < 0.01, P < 0.05), and the concentration of SOD and TNF-alpha of MEE in the mucous fluid group was higher than that in the serous fluid group (P < 0.01, P < 0.05). SOD Level of MEE was positively correlated with TNF-alpha (r = 0.587, P < 0.01). CONCLUSION: SOD, IL-6, IL-8 and TNF-alpha might be important mediators in MEE secretion in SOM. IL-6 and IL-8 might participate in the defensive reaction of organism during the early stages of SOM. SOD and TNF-alpha might be closely related to the persistence of SOM, and might be mediators of mucin secretion in SOM. PMID- 12541691 TI - [Bcl-2, p53 expression and Epstein-Barr virus infection in primary nasopharyngeal carcinoma]. AB - OBJECTIVE: To observe bcl-2, p53 gene expression and Epstein-Barr virus (EBV) infection and their relationship with clinicopathologic factors in primary nasopharyngeal carcinoma (NPC). METHOD: p53, bcl-2 proteins were assayed with immunohistochemical technique and EBV were assayed with polymerase chain reaction in 54 cases of NPC and 13 cases of chronic inflammation of nasopharyngeal epithelium (CINE). RESULT: The expression of p53, bcl-2 proteins and EBV were found respectively in 1(7.7%), 3(23.1%) and 1(7.7%) of 13 CINEs, which were significantly lower than those in 54 NPCs with respectively 41(75.9%), 48(88.9%) and 47(87%) (P < 0.01). In 39 cases of 54(72.2%) NPCs, we could find simultaneous p53, bcl-2 expression and EBV infection, furthermore, p53 and bcl-2 expression were significantly positively correlated with each other (r = 0.78, P < 0.01). p53 gene expression was significantly correlated with cervical lymph node metastasis of NPCs (P < 0.01). The expression of p53 in stage II, III, IV (92.9%, 78.9% and 100% respectively) were significantly higher than that in stage I (38.5%) (P < 0.01). CONCLUSION: The expression of p53, bcl-2 gene and EBV infection are the useful factors in the diagnosis of NPC, the overexpression of p53 is related with the stage and cervical lymph node metastasis of NPC, all the three play important roles in the pathogenesis of nasopharyngeal carcinoma. PMID- 12541692 TI - [The 5-year death analysis in patients with nasopharyngeal carcinoma treated with radiotherapy]. AB - OBJECTIVE: To study the main death cause of nasopharyngeal carcinoma (NPC) treated with radiotherapy for 5 years. METHOD: An analysis had been made to evaluated the 5-year death in 165 patients with NPC treated with radiotherapy. RESULT: The death correlates strongly with the clinical stage, type and radiation dose (P < 0.05). CONCLUSION: The distant metastasis and local recurrence are the main death cause of NPC. PMID- 12541693 TI - [The influence of labyrinthectomy on the expression of NMDA R1 mRNA]. AB - OBJECTIVE: To study the effect of labyrinthectomy on the expression of NMDAR1 (N methyl-D-aspartate receptor subunit R1). METHOD: The in situ hybridization was used to observe the expression of NMDA R1mRNA post-unilateral labyrinthectomy and investigate its effect on vestibular compensation. RESULT: 20 h post-unilateral labyrinthectomy, the NMDA R1mRNA positive neurons in the medial vestibular nucleus increased significantly compared with the unlesioned side(P < 0.01). 40 h 6 d, the positive cells decreased gradually. 6 d post lesion, the positive neuron counts of the lesioned side had no significant difference compared with those of the un-lesioned side (P > 0.05). CONCLUSION: The changing trend of the expression of NMDA R1mRNA in the medial vestibular nucleus is related to the recovery of the vestibular static symptoms after unilateral labyrinthectomy. It is suggested that the modulation of the efficacy of the NMDA R1 play an important role in the compensation of the static symptoms. PMID- 12541694 TI - [The application of multi-pulse stripping amperometric detection to capillary electrophoresis]. AB - There are some drawbacks in amperometric detection in capillary electrophoresis since there is influence of high voltage from electrophoretic instrument. Three electrodes of electrochemical detection system and electrochemical system of capillary electrophoresis are separately placed in capillary outlet buffer reservoir, which can lead to a reduction in the influence of high voltage in the great degree. A carbon fiber with diameter of 7 microns is used as working electrode, and placed just at the separation capillary outlet. The multi-pulse stripping amperometric detection has 1.1 s deposition step and several stripping steps with 0.02 s, which can increase the detection amperometric current and identify the species of each separated component in some classes by different pulse stripping potentials. This detection technique was applied in the separation of Cu, Zn, Pb, Tl and Cd of 25 mg/L by the capillary electrophoresis, and separation efficiency is over 60 thousands theoretical plates. The limit of the multi-pulse stripping amperometry detection was 0.10 microgram/L for cadmium by using water plug field amplified injection. PMID- 12541695 TI - [Conical column end for wide bore packed capillary liquid chromatographic column]. AB - Wide bore (> or = 0.5 mm i.d.) packed capillary liquid chromatographic columns with conical end were designed and evaluated. It was found that the column efficiency was 30%-50% higher than that of the packed capillary columns with conventional end connection at optimal flow rate, and was twice as much as that at reduced flow rate of 10-15. The chromatographic peak symmetry and the van Deemter curve of the novel shape column were also improved substantially. Fast analysis of PAHs is demonstrated. PMID- 12541696 TI - [Determination of nitrite and nitrate in rat nervi ischiadicus by high performance liquid chromatography]. AB - To study the effect of nitric oxide on the diabetic neuropathy, a quantitative method for the determination of nitrite(NO2-) and nitrate(NO3-) in rat nervi ischiadicus by high performance liquid chromatography(HPLC) with UV-VIS detection was developed. In the range of 1 mumol/L-25 mumol/L, correlation coefficient between concentration and peak area for NO2- and NO3- was more than 0.991. The minimum detection limits for both ions were 0.2 mumol/L and 0.5 mumol/L, respectively. Relative standard deviation(RSD) of inter-day and intra-day was less than 14%. The effects of different acetonitrile concentration and pH of mobile phase on separation as well as the stability of both salty ions were studied. In comparison with normal group, the levels of NO2- and NO3- for diabetic groups were lower. Measured mean values were, in turn, as follows: normal rats, (2.62 +/- 0.38) mumol/L and (24.43 +/- 5.61) mumol/L (n = 8); diabetic-controlled rats (saline-given only), (2.24 +/- 0.20) mumol/L and (17.15 +/- 7.79) mumol/L (n = 6); diabetic rats treated with bovine colostrum insulin like growth factor(BctIGF-1) or with recombinant human insulin-like growth factor (rhIGF-1), (2.55 +/- 0.36) mumol/L and (6.12 +/- 2.14) mumol/L (n = 5) and (1.46 +/- 0.08) mumol/L and (9.95 +/- 3.34) mumol/L (n = 7), respectively. PMID- 12541697 TI - Rapid analysis of terpene lactones in extract of Ginkgo biloba L. by high performance liquid chromatography. AB - A new rapid analytical method was developed for the quantification of terpene lactones (bilobalide(BB) and ginkgolide A, B, C, J) in extract of Ginkgo biloba L. (EGb) using a liquid-liquid solvent extraction procedure followed by high performance liquid chromatography. EGb was dissolved in 30% ethanol and extracted with ether. After evaporation, the residue was then determined by HPLC on a C18 column with methanol-water-orthophosphoric acid (25:75:0.1, V/V) as eluent and refractive index (RI) detection. Results showed that the excellent sample clean up procedure is more simple and specific, and saves more time (less than 20 min) than any other methods that have been reported, and also leads to high recoveries (> 99.0%) and low RSDs (< 2.0%). The reproducible method is regarded to be very useful for evaluating the quality of extract of Ginkgo biloba L. PMID- 12541698 TI - [The evaporative light-scattering detection technology]. AB - The theory, principles of operation and theoretical investigation of evaporative light-scattering detector (ELSD) are reviewed with 45 references. The applications of ELSD in HPLC on lipids, surfactants, pharmaceutical compounds and others are introduced. PMID- 12541699 TI - Advances in capillary chromatography. AB - Capillary columns are used in both capillary liquid chromatography and capillary electrochromatography. The design for capillary liquid chromatography is discussed in comparison with capillary gas chromatography. The difference of diffusion coefficient in gas and liquid phase is a key role. The study for obtaining a high performance capillary liquid chromatography is discussed. Capillary electrochromatography is recently interesting for its instinct ability to realize a high performance chromatography. Capillary electrochromatography with and without pressurized flow is reviewed briefly. Instrumentation for capillary electrochromatography with pressurized flow is discussed. The port of splitting, and gradient elution of both solution and potential are described. The new findings of both the variation of column resistance and capacity factor according to the value of applied electric voltage are also discussed. PMID- 12541700 TI - [Recent advances in the ion chromatographic stationary phases]. AB - Four types of the ion chromatographic stationary phases are discussed. 96 references are cited, among which 63 are published after the year of 1995. PMID- 12541701 TI - [A simple method for preparing reversed-phase electrochromatographic column and its evaluation]. AB - The packed column was prepared with a simple method by combination of vacuum filling and pressure compressing. The reproducibility of efficiency and retention time was evaluated. The mixture of thiourea and three aromatic compounds was used. RSD values of retention time were smaller than 2.2% and RSD values of efficiency were smaller than 7.5%. The influence of voltage, phosphate concentration, acetonitrile volume fraction and column temperature on the electroosmotic flow and capacity factor of neutral compounds was investigated. Acetonitrile concentration had significant effect on the electroosmotic flow and capacity factor of neutral compounds. High concentration of acetonitrile improved the elution ability. Low concentration of acetonitrile improved the resolution. Voltage, phosphate concentration, and column temperature also had effects on the electroosmotic flow and capacity factor of neutral compounds, but the effects were smaller than the concentration of acetonitrile. These results gave a guide to select separation conditions in capillary reversed-phase electrochromatography. PMID- 12541702 TI - [Determination of the active components in Chinese herb Aloe vera L. var. chinensis (Haw.) Berger by capillary zone electrophoresis]. AB - It has been proved that the Chinese herb Aloe vera L. var. chinensis (Haw.) Berger as well as its active components showed many important pharmacology activities. In order to find an easy and low-cost method to control the quality of the herb, a CZE method for the determination of the active components aloin and aloe-emodin in Aloe vera L. var. chinensis (Haw.) Berger was developed in this work. Under the buffer conditions of 24 mmol/L phosphate (pH 10.52), applied voltage of 15 kV and detector wavelength of 254 nm, baseline separation of the active compounds in Aloe vera L. var. chinensis (Haw.) Berger was achieved and the active components were quantitatively analyzed. The linear calibration equations of the two components are: Y= -0.140 + 57.2X (r = 0.997) for aloin and Y = -0.393 + 1.08 x 10(2) X (r = 0.999) for aloe-emodin respectively. In addition, the effects of buffer pH value and organic modifier on the migration behavior of the solutes were also investigated. PMID- 12541703 TI - [Determination of sialic acids in serum of lung cancer with ultrafiltration capillary electrophoresis]. AB - A method for the determination of sialic acids in serum with ultrafiltration capillary electrophoresis is described and the operation was optimized. Sialic acids were directly separated and analyzed with UV detection at 195 nm and without pre-or post-column derivatization. The recovery of N-acetylneuraminic acid (NANA) was 92.6%, the concentration and mass detection limit of NANA were 9.6 mumol/L and 39 fmol respectively. This method was used for the determination of NANA level in the serum of 11 lung cancer patients and 30 normal adults. The results showed that the average concentration of NANA in the serum of patients was much higher than that of normal adults with P < 0.001. The results were also compared with those obtained by the traditional colorimetric method, with good linear relationship of r = 0.983 at n = 10. It is concluded that the method described in this paper is simple and sensitive, and is suitable for basic research and clinical applications to malignant tumors. PMID- 12541704 TI - [Determination of relative molecular mass distribution of electro-oxidative degradation products of lignosulfonate by gel permeation chromatography]. AB - The relative molecular mass and relative molecular mass distribution of electro oxidative degradation products of sodium lignosulfonate were determined using a Waters ALC/GPC 244 apparatus comprising a mu-Ultrahydrogel 250 column and a differential refractometer index detector. The influences of the mobile phases on the elution behaviors of the samples are discussed. It is confirmed through a series of experiments that the eluent of a neutral aqueous solution containing low concentration of electrolyte could separate the components of the degradation products of lignosulfonate. The best results were obtained by using 0.05 mol/L LiCl solution with pH 6.5 as the eluent. PMID- 12541705 TI - [HPLC determination of PAHs from electric power plant by UVD-FLD connected in series]. AB - An HPLC method of UVD-FLD connected in series was used to detect naphthalene, biphenyl, fluorene, phenanthrene, anthracene, fluoranthene, 2-methyl-anthracene, chrysene, benzanthracene, benzfluoranthene [b], benzfluoranthene[k], benzopyrene PAH compounds in electricity plant environment. A CLC-ODS column (150 mm x 6.0 mm i.d.) with acetonitrile-water(85:15, V/V) as mobile phase and UV detector set at 250 nm and fluorescent detector set at lambda ex295 nm and lambda em427 nm were used. These 12 PAHs can be separated completely by this method. The earlier eluted eight PAHs were detected by UVD and the rest were detected by FLD. The minimum detectable concentrations of the 12 PAHs were 0.002 mg/L-0.1 mg/L. Satisfactory results were obtained in the analysis of PAH compounds in waste water, flue gas and flyash from several electric power plant in Hunan Province. PMID- 12541706 TI - [Simultaneous determination of Cd2+, Pb2+, Cu2+ and Zn2+ by reversed-phase high performance liquid chromatography]. AB - HPLC was used in the investigation of the reaction behavior of Cd2+, Pb2+, Cu2+ and Zn2+ with mesotetrakis (4-hydroxy-phenyl)porphine (THPP) and chromatographic behavior of their complexes. The reaction and chromatographic separation parameters were optimized and a method for the simultaneous determination of Cd2+, Pb2+, Cu2+ and Zn2+ by reversed-phase high performance liquid chromatography with THPP as pre-column derivatization reagent was established. The metal-THPP complexes formed in 0.1 mol/L borate buffer (pH 10.5) by heating in 60 degrees C water bath for 20 min and were extracted into ethyl acetate benzene mixture. The metal-THPP complexes and THPP were separated on a Prodigy ODS, 5 microns, 250 mm x 4.6 mm i.d. column with methanol-0.05 mol/L borate buffer(90:10, V/V) (pH 8.0) as eluent and detected at 437 nm (Cd2+ and Zn2+), 465 nm (Pb2+) and 416 nm (Cu2+). The detection limits of the four metal ions in 5 microL injection solution were Cd2+ 0.02 ng, Pb2+ 0.02 ng, Cu2+ 0.02 ng, and Zn2+ 0.12 ng, and the linear ranges were Cd2+ 8 ng-1.5 micrograms, Pb2+ 8 ng-3.0 micrograms, Cu2+ 8 ng-5.0 micrograms, and Zn2+ 50 ng-10 micrograms in 10 mL reaction solution. The intra-day repeatability (as RSDs) was in the range of 2.8% 4.8% and the inter-day reproducibility was in the range of 3.7%-9.7%. PMID- 12541707 TI - [Determination of six ginsenosides in Panax species by high performance liquid chromatography]. AB - A method for the determination of six major ginsenosides in panax species by high performance liquid chromatography (HPLC) with UV detection at 203 nm is described. The six major ginsenosides, which were reported to account for more than 90% of the total ginsenosides content of the panax species' root, were successfully separated using gradient HPLC. The column to be used was a Cosmosil C18 reversed-phase, 4.6 mm i.d. x 250 mm, 5 microns (Japan), connected with a C18 guard column and the mobile phase was acetonitrile and water. The calibration curves of the six ginsenosides were all linear over the range of 25 mg/L-300 mg/L with good correlation coefficients (> 0.99). The recoveries for the ginsenosides were all above 80%. For real samples and recovery study, the major ginsenosides were ultrasonically extracted by methanol. The extracts were evaporated to 1 mL, under vacuum, at about 50 degrees C and then diluted by methanol. Just prior to HPLC analysis, the sample solutions were filtered using a C18 cartridge. This method has been successfully applied to the analysis of four commercial panax samples. PMID- 12541708 TI - [Determination of three principal components in chuanjing tablets by using high performance liquid chromatography]. AB - Uniform design method was employed to optimize the mobile phase of HPLC in order to determine simultaneously three principal components, theophylline, amobarbital and methylephedrine hydrochloride, in Chuanjing tablet, a compound preparation for asthma and cough. The stationary phase was ODS and the optimal mobile phase composition was V (0.015 mol/L phosphate buffer containing 0.3% triethylamine, pH 4.9): V (methanol) = 35:65. The detection was performed at 215 nm and the sensitivity was time programmed for simultaneous determination of minor and major components. Caffeine was selected as the internal standard. A baseline separation was achieved within 10 min. The linear ranges for theophylline, amobarbital and methylephedrine hydrochloride were 0.03 g/L-0.20 g/L, 7.5 mg/L-50.0 mg/L and 7.5 mg/L-50.0 mg/L, recoveries 99.7%-102.6%, 98.5%-100.2% and 98.0%-102.7%, inter-day RSDs 0.23%-1.2%, 0.35%-2.5%, 0.33%-1.6% respectively. This HPLC method is rapid and accurate, and suitable for the quality control of the preparation. PMID- 12541709 TI - [Determination of EDTA residue in canned food products by RP-HPLC]. AB - The homogenized sample is diluted with water. After adding copper(II) chloride solution and 20 mg of ascorbic acid, dilute the sample to the volume with water. After thorough mixing and microfiltration over a 0.45 microns filter, the filtrate is ready for injection into the HPLC system and detected at 254 nm. The 125 mm x 4 mm i.d. chromatographic column is packed with Hypersil ODS 5 microns, eluted with a mobile phase of water-methanol (80:20, V/V) containing 20 mmol/L tetrabutyl ammonium bromide, 0.03 mol/L sodium acetate buffer (pH 4) at a flow rate of 0.8 mL/min. The injection volume is 20 microL. The response value was linear between 10 mg/L-400 mg/L. The recovery was 95.5%-98.9%. The RSD was 0.82% 1.32%. PMID- 12541710 TI - [Study on the analysis of NO2-, Br- and NO3- by ion chromatography in the presence of high mass concentration of chloride]. AB - The effects of high mass concentration chloride on the determination of NO2-, Br- and NO3- were studied on three different analytical columns by ion chromatography. The variations of retention behavior of these anions were caused by self-elution effect and on-column eluent change in the presence of high concentration chloride. When their peaks overlapped with chloride, the deviations of the quantity of weak acid anions were caused by anion-proton interactions within the suppressor. However, the strong acid anions and the other weak acid anions, not overlapped by chloride, were not affected. The linear ranges of these anions in different concentrations of chloride and the maximum chloride concentration allowed to be measured by peak height quantitative method were examined. PMID- 12541711 TI - [Analysis of headspace constituents of Gardenia flower by GC/MS with solid-phase microextraction and dynamic headspace sampling]. AB - The headspace constituents of fresh Gardenia flower were investigated by GC/MS. The headspace volatiles were sampled by solid-phase microextraction (SPME) and dynamic headspace sampling (DHS). SPME sampling was conducted with 100 microns PDMS fiber at 28 degrees C for 60 min. In DHS sampling, purified nitrogen was used as purging gas with a flow rate at 80 mL/min for 120 min. Tenax GR(20 mesh 40 mesh) was used as adsorbent and the volatiles were eluted by ether, and concentrated to 0.5 mL for GC/MS analysis. A Supelco-wax capillary column (30 m x 0.25 mm i.d. x 0.25 micron df) was employed in GC/MS analysis. Initial oven temperature was kept at 45 degrees C for 2 min, then raised to 250 degrees C at 4 degrees C/min, and kept at 250 degrees C for 10 min. According to SPME-GC/MS analysis, the main compounds in headspace of fresh Gardenia flower included farnesene(64.86%), cis-ocimene(29.33%), linalool(2.74%), cis-3-hexenyl tiglate(1.34%), methyl benzoate(0.25%). Results obtained from SPME and DHS sampling were also compared. In this study, SPME afforded a simpler and more sensitive sampling method, and much more accurate information about headspace volatiles of Gardenia flower. PMID- 12541712 TI - [Separation and determination of glibenclamide in xiaotangling tablets by micellar electrokinetic capillary chromatography]. AB - A high performance capillary electrophoresis method has been developed for separation and quantitative analysis of glibenclamide in Xiaotangling tablets. Electrophoretic conditions were as follows: micellar electrokinetic capillary chromatography (MECC) mode, trimethoprimum as internal standard, uncoated fused silica capillary (34.8 cm/39.5 cm(effective/total length), 50 microns i.d.), 25 mmol/L borax-30 mmol/L SDS(pH 9.0), applied voltage 17 kV, (+)-->(-), temperature 28 degrees C, detection wavelength 228 nm, pressure injection 68.95 kPa.s. Glibenclamide was separated successfully from other components within 14 minutes under the optimum conditions, and the relative peak area of glibenclamide increased linearly with the increase of its concentration within the range of 25 mg/L-275 mg/L. The recovery was (100.6 +/- 1.4)%. The method is simple, rapid and well reproducible, and can be used as a reliable tool for the quality control of Chinese traditional medicine containing glibenclamide. PMID- 12541713 TI - [The determination sorbitol and carbohydrates in nane fruit by high performance liquid chromatography]. AB - A high performance liquid chromatographic method for the determination of carbohydrates and sorbitol in Nane fruit is described. The chromatography was carried out on a mu-Spherogel carbohydrate column(4.6 mm i.d. x 300 mm) (at 85 degrees C) using water as mobile phase and refractive index detector. The average recovery of carbohydrates were 75.00%-88.37% and correlation coefficients were 0.9709-0.9999. The relative standard deviations for intra-day were 5.10% for sucrose, 3.15% for glucose, 3.03% for fructose and 4.84% for sorbitol respectively (n = 5). The accumulations of soluble carbohydrates(including fructose, glucose, sorbitol and sucrose) in flesh showed their characteristic patterns in different special phase of fruit growth. PMID- 12541714 TI - [Determination of 4 antiseptics and 6 sunscreen agents in cosmetic products by reversed-phase high performance liquid chromatography]. AB - A method for the quantitative determination of 4 antiseptics (methyl p hydroxybenzoate, ethyl p-hydroxybenzoate, n-propyl p-hydroxybenzoate, n-butyl p hydroxybenzoate) and 6 sunscreen agents (2-hydroxy 4-methoxybenzophenone-5 sulfonic acid, 2-hydroxy 4-methoxybenzophenone, phenyl salicylate, 2-ethylhexyl 4 dimethyl aminobenzoate, 2-ethylhexyl 4-methoxycinnamate, 2-ethylhexyl salicylate) in cosmetic products was developed successfully. It was based on a reversed-phase high performance liquid chromatographic separation under the conditions of isocratic elution with a mixture of methanol-THF-water (30:100:100, V/V) by using a column packed with 10 microns YWG-C18, and the UV detection at 254 nm and 310 nm. The recoveries(n = 6) ranged from 87.2% to 106.5%, and the relative standard deviations(n = 6) were from 1.2% to 3.3%. PMID- 12541715 TI - [Determination of beberine hydrochloride and puerarin in tangwei capsules by RP HPLC of gradient elution method]. AB - A reversed-phase high performance liquid chromatographic gradient elution method was developed for the determination of beberine hydrochloride and puerarin in Tangwei capsules. Separation was obtained by using Hypersil ODS (150 mm x 4.6 mm i.d., 5 microns) column and mobile phase of methanol and acetic acid, triethylamine water solution. Detection was performed with UV detector at 230 nm. The column temperature was 35 degrees C. The calibration curves were linear within the range of 0.18 microgram-1.66 micrograms for beberine hydrochloride (n = 5, r = 0.9999) and 0.09 microgram-0.89 microgram for puerarin (n = 5, r = 0.9998). The recovery and RSD of beberine hydrochloride were 98.2% and 1.6% and those of puerarin were 97.5% and 1.8%, respectively. PMID- 12541716 TI - [Determination of brucine in seed dressing agents by high performance liquid chromatography]. AB - An HPLC method for the determination of brucine in seed dressing agents was investigated. The sample solution was separated on a Spherisorb C18 column(4.6 mm i.d. x 200 mm, 5 microns) with CH3OH-H2O-CTAB (88:12:0.04, m/m) as the mobile phase and detected at 254 nm. Calibration curve of brucine was obtained for the concentration range of 0.01 g/L-0.25 g/L. The linear regression equation was A = 11,485,814 rho + 21,229, r = 0.9997. The RSD was 1.2%(n = 11). The results show that this method is simple, specific and accurate. PMID- 12541717 TI - [Preparative GC separation and identification of methyl 2-methyl-3-furyl disulfide flavour]. AB - On 4% FFAP + 1% H3PO4 Chromosorb W/AW-DMCS (80 mesh-100 mesh)column, the optimized separation of methyl 2-methyl-3-furyl disulfide flavour was obtained. By using automatic preparative attachment model App-5 to GC-9A, the content of methyl 2-methyl-3-furyl disulfide flavour was raised from about 77% to 99.5%. The product was identified by IR, MS and 1HNMR. PMID- 12541718 TI - [Micro determination of acrylonitrile in ambient air by gas chromatography]. AB - Acrylonitrile in ambient air was collected by Tenax GC and then thermo-desorbed. It was separated on GDX-502 chromatographic column from other pollutants and determined by flame-ion ization detector. The minimum detectable concentration was 0.01 mg/m3 when 2 L ambient air was collected. The recoveries were 85.6% 105.4% and RSDs were 4.5%-6.2%. PMID- 12541719 TI - [Clinical comparison of bilateral neck dissection for supraglottic carcinoma]. AB - OBJECTIVE: In order to rise the survival rate and quality of life, the patients with bilateral neck dissection was investigated in supraglottic carcinoma. METHOD: 76 cases were retrospectively analyzed. In terms of clinical diagnosis, 33 cases who had bilateral neck metastasis had received simultaneously bilateral neck dissection. 43 patients first diagnosed unilateral neck metastasis got subsequent bilateral neck dissection. All of the patients were pathologically diagnosed. RESULT: The 3 and 5 year survival rates, in the 33 cases who had received simultaneous bilateral neck dissection, were 81.5% (22/27) and 61.5%(16/26) respectively, while in the other 43 cases, was 69.2%(27/39) and 27.8%(10/36) respectively. CONCLUSION: It is necessary for those patients who have suffered from supraglottic carcinoma with bilateral neck metastasis to have simultaneous bilateral neck dissections. Meanwhile, the preservation of bilateral internal jugular veins is the best choice, at least one should be preserved. PMID- 12541720 TI - [Acoustic analysis in patients with trauma to unilateral cricoarytenoid joint by computer technique]. AB - OBJECTIVE: To investigate the acoustic characteristics of unilateral cricoarytenoid joint trauma and evaluate the effect of acoustic analysis technique on the diagnosis and treatment of this disease. METHOD: The voice signals of sustained vowel [a] were measured using a micro-computer with Dr. Speech software in 50 healthy adults and 30 patients with unilateral cricoarytenoid joint trauma. The acoustic parameters (jitter, shimmer and NNE) and spectrographic characteristics (harmonic waves, formants and noise) were analyzed. The acoustic changes before and after the treatment of arytenoid motion were observed and compared. RESULT: All acoustic parameters were significantly increased in trauma of unilateral cricoarytenoid joint. The pathologic spectrograph showed decrease or loss of harmonic waves and formants, and increase of noise, in middle and high frequencies. There was a relationship between NNE and the position of the vocal cord. After the treatment of arytenoid motion, 33% patients were cured and had normal acoustic parameters and spectrograph. 47% were improved with reduced parameters and improved spectrograph. The acoustics in 20% cases didn't change at all after treatment. The acoustic parameters were significantly decreased after the treatment of arytenoid motion. CONCLUSION: The present study suggested that the acoustic parameters may estimating the degrees of voice lesion as objective standards and NNE may judge the degree of glottic closure. Both acoustic parameters and spectrograph are of value in the diagnosis, treatment effect and prognosis observation to cricoarytenoid joint trauma. PMID- 12541721 TI - [The relationship between content of substance P, VIP in pharyngeal tissue and narrow pharyngeal cavity of patients with OSAS]. AB - OBJECTIVE: To investigate the relationship between content of substance P, VIP in the pharyngeal tissue and narrow pharyngeal cavity of patients with OSAS. METHOD: By using semi-quantitative immunohistochemical method, resected pharyngeal tissue from 30 patients with OSAS and 12 normal health adult controls were investigated with rabbit anti-substance P and rabbit anti-VIP. RESULT: SP and VIP were detected in tribution within the epithelium or between epithelium cells, around the pharyngeal glands and gland ducts, around endothelium cells and smooth muscle of blood vessels. SP and VIP levels in pharyngeal tissue of the patients with OSAS were increased than those of the controls. CONCLUSION: Narrow of pharyngeal cavity with OSAS was relative to edema of pharyngeal tissue induced by increased SP and VIP in pharyngeal tissue. A neurogenic inflammation was one of the factors of OSAS. PMID- 12541722 TI - [Differentiated microcarcinoma of the thyroid gland in 45 cases]. AB - OBJECTIVE: To offer therapeutic management for patients with microcarcinoma of the thyroid gland (TMC). METHOD: Forty-five patients with TMC were treated from 1993, including 10 males and 35 females. Homolateral total thyroidectomy + contralateral subtotal thyroidectomy + homolateral functional neck dissection was taken as routine. RESULT: Among 168 patients with differentiated carcinoma of the thyroid gland, 45 patients were TMC (26.78%). Homolateral metastasis of lymph node presented in 11 (24.44%) of the TMC patients. No operation complications. All patients were followed up. No recurrence, no death. CONCLUSION: The lymph node metastasis appears to be high. Homolateral total thyroidectomy + contralateral subtotal thyroidectomy + homolateral functional neck dissection was effective and essential. PMID- 12541723 TI - [The selection of surgical technique for differentiated thyroid carcinoma]. AB - OBJECTIVE: To evaluate the optimal extent of primary thyroidectomy and neck dissection in the patients with differentiated thyroid carcinoma (DTC). METHOD: Retrospective analysis of 66 cases of DTC. Lobectomy for tumor side plus subtotal lobectomy for another side was performed in 49 cases, lobectomy with isthmectomy was performed in 7 cases, subtotal lobectomy for one side plus partial lobectomy for another side was performed in 6 cases, total thyroidectomy was performed in 4 cases. Unilateral functional neck dissection for another side was performed in 1 case and unilateral traditional neck dissection in 5 cases. RESULT: The 5 and 3 years survival rates were 94% and 96%. All 4 cases operated with total thyroidectomy suffered hypothyroidism and 2 of them had hypoparathyroidism postoperatively. There was no hypothyroidism and hypoparathyroidism for other cases. No recurrent nerve paralysis occurred in this group. CONCLUSION: We recommend lobectomy for tumor side plus subtotal lobectomy for another side for most of DTC. Selective neck dissection is not necessary for N0 cases unless the patient is in high risk group. PMID- 12541724 TI - [Clinical study of decompression of optic nerve through combined orbit, ethmoid and sphenoid approach]. AB - OBJECTIVE: To improve the level of diagnosis and treatment of optic nerve trauma at the canalicular segment. METHOD: Fifty-two patients with optic nerve trauma at the canalicular segment received decompression of optic canal through combined orbit, ethmoid and sphenoid approach. RESULT: The visual improvement rate of all 52 patients was 42.3%, 73.9% in 23 patients with secondary optic trauma and 17.2% in 29 patients with primary optic trauma. Patients's vision covered from 0.05 to 0.6 after six months. CONCLUSION: The advantages of decompression of optic nerve through combined orbit, ethmoid and sphenoid approach include easily operation and location, short approach, wide visual field and clear anatomy mark, so it is very safe and suited for those patients with dysplasia and osteosclerosis of ethmoidal or sphenoidal sinus. Orbital CT scan facilitates the diagnosis of optic canal fracture without false positive results. The curative effect is associated with the type of optic nerve trauma and the chance of surgery. Amaurosis shortly after trauma is a dangerous factor of poor prognosis, but it isn't impossible to improve vision if patients receive operation as early as possible. PMID- 12541725 TI - [Study on relationship between the reflux laryngitis and gastroesophageal reflux disease]. AB - OBJECTIVE: To investigate the relationship between reflux laryngitis and gastroesophageal reflux disease (GRED). METHOD: 130 Patients of refractory chronic laryngitis with upper endoscopy and ambulatory 24-hr esophageal pH-metry. The patients of gastroesophageal reflux disease were divided into two groups. Group A was treated with antacids and motile medications. Group B was control. RESULT: 45 (34.6%) gastroesophageal reflux disease were found in 130 cases. 21 cases(91.3%) were healed in group A. 3 of 22 cases (13.6%) were healed in control (P < 0.01). CONCLUSION: The gastroesophageal reflux disease was an important cause of the reflux laryngitis which symptoms and signs can be healed by antacids and motile medications. PMID- 12541726 TI - [The clinical investigation of bacteriology of chronic suppurative otitis media]. AB - OBJECTIVE: To understand bacteriology of chronic suppurative otitis media (CSOM) and drug sensitivity of isolated bacteria for better treatment of CSOM. METHOD: Secretions of 92 samples with chronic suppurative otitis media were collected and cultured for aerobic and anaerobic bacteria respectively. Drug sensitivity test of isolated bacteria was done. RESULT: Aerobic bacteria was found in 75 of 92 samples (81.5%), the main pathogens were pseudomonas aeruginosa, aureus staphylococcus and bacillus proteus, and their sensitive rates for ciprofloxacin were 89.0%, 88.2% and 85.0% respectively. Anaerobic bacteria was found in 10 samples (10.9%) and the relative sensitive drug was cefoperazone sodium. CONCLUSION: Pseudomonas aeruginosa, aureus staphylococcus and bacillus proteus are the main pathogens of chronic suppurative otitis media, and their sensitive rates for ciprofloxacin are high. PMID- 12541727 TI - [Nasal cavity dimensions in the infants measured with acoustic rhinometry]. AB - OBJECTIVE: To investigate the range of normal value of minimal cross-sectional area (MCA) of the nasal cavity, distance of minimal cross-sectional area from the nostril (DCA), total nasal cavity volume(NV) and total nasopharyngeal cavity volume (NPV) of healthy infants. METHOD: One hundred and seventy-five healthy infants and one hundred and eight infants suffered from acute upper respiratory tract infection were measured with acoustic rhinometry. RESULT: The range of total MCA of healthy infants was 0.2-0.5 (cm)2, DCA was 1.93-2.47 cm, NV was 2.69 4.75 cm3 and NPV was 3.83-8.92 cm3. Comparing healthy infants with infants suffered from acute upper respiratory tract infection, there all were significant differences in MCA, DCA and NV. CONCLUSION: Acoustic rhinometry suit objective assessment of the nasal airways in infants. It is demonstrated that these data can provide available information for the study of nasal physiology and pathophysiology, well as for the diagnosis and judgement of therapeutic effectiveness of nasal diseases. PMID- 12541728 TI - [Experimental studies on antitumor effect by the new recombinant tumour necrosis factor and cyclophosphamide in nude mice bearing laryngeal carcinoma]. AB - OBJECTIVE: We aimed at finding ideal biological treatment and regime for laryngeal carcinoma. METHOD: Nude mice model bearing laryngeal carcinoma were established using laryngeal carcinoma cell line (HEP-2). New recombinant human tumor necrosis factor (NrhTNF-alpha) and cyclophosphamide were administered locally or systemically, single or in combination. The microstructural and ultra micro-structural changes of carcinoma after administration of NrhTNF-alpha were observed under light and electron microscopes. RESULT: 1. After treatment, the tumor showed hemorrhage, necrosis, regression, the survival time was prolonged. 2. More significant antitumor effect was observed when locally administered NrhTNF-alpha than intravenously administered of NrhTNF-alpha. 3. There is a notable synergistic antitumor effect by the combination use of NrhTNF-alpha and cyclophosphamide. 4. Microscopy showed that cell changes mostly took place in plasma especially in mitochondria and endoplasmic neticula. The mitochondria were highly bloated. Besides, some nuclei were condensed and fragmented resulted in cell death. CONCLUSION: These studies provide experimental basis for biological treatment of laryngeal carcinoma. PMID- 12541729 TI - [A survey of pollens in Chongqing]. AB - OBJECTIVE: In order to investigate the variety of the airborne pollen in Chongqing. METHOD: The diffusions pattern of airborne pollens in two areas of Chongqing was investigated for one year from October 1999 to September 2000. The slides were exposed daily through one year. RESULT: The peak of spreading was in March in spring and in Autumn from September to October. The test on the pollen patients shows that the main resulting pollen is in Autumn. CONCLUSION: The airborne pollen peak in Chongqing in spring prone to the influence of temperature, wind power and rain amount. PMID- 12541730 TI - [Study on the rule of solvent strength in reversed-phase liquid chromatography]. AB - Based on the physical meanings of Z (total moles of the released solvent at the contact region between solute and stationary phase), and j(the slope of the linear plot of lg I versus Z, a constant relating to the affinity of organic solvent to the stationary phase) of stoichiometric displacement model for retention (SDM-R) of solute in liquid chromatography(LC), both Z and j could be employed to characterize solvents, therefore, Z value only can be used as a characterization parameter for fixed solute and stationary phase. However, the parameter j is independent of solutes. As long as phase is given, j can be employed as a new characterization parameter of organic solvent strength in LC. With the homologues of methanol, ethanol and n-propanol as organic solvents in mobile phase and n-alkylalcohols as solutes, both the SDM-R and Snyder's empirical equation were tested, and compared with the corresponding parameters obtained from Snyder's empirical equation. With S [the slope of the linear plot of lg k' (k' is capacity factor) vs psi(volume fraction of organic solvent in mobile phase)] and lg kw0(the intercept of this plot), it was found that the two linear plots with the SDM-R, lg k' vs the composition of mobile phase and lg I vs Z were better than the corresponding plots with Snyder's empirical equation. The results indicate that Z, lg I and j decrease with the increase in the molecular size of the organic solvent of the homologue. This fact indicates that the solvent strength in LC depends on the size of solvent molecules. And the greater the solvent molecules, the stronger the solvent strength is. Furthermore the solvent strength was found to follow homologous rule. The j value may become a new parameter for the characterization of organic solvent strength in LC. PMID- 12541731 TI - [Prediction and estimation on molar response values of alkanes by using molecular path vector]. AB - A new method based on a novel molecular topological index vector, called the molecular path vector (MPV), of alkane molecules is proposed and employed for estimation and prediction of the molar response values of various alkanes. The novel MPV, p = (P1, P2, P3, P4, P5, P6, P7, P8, P9, P10)', which derived directly from the interaction terms of molecular graph, is used to characterize well molecular structures of all alkanes from one through ten or eleven carbon atoms. It showed that there exists very good correlation between the MPV elements and molar response values on both FID and TCD detectors in classical gas chromatography. Based on the given calibration set with different sample numbers and by using the practical multiple linear regression, the quantitative structure response relationship (QSRR) equations, for the molar response values (SM) on both FID and TCD, are respectively given as follows: SM(FID) = 15.4004881 + 17.9905995 X1 - 0.1652116 X2 - 0.6974103 X3 - 0.8452390 X4 - 0.2671000 X5 - 1.5657273 X6 + 0.0944440 X7, n = 50, m = 7, r = 0.9976, ST = 26.132, SR = 1.965 1, Ev = 99.72%, RMS = 1.801, F = 1231.71 SM(TCD) = 11.9946996 + 29.1490916 X1 - 4.7451669 X2 - 3.7673385 X3 - 1.4948330 X4 - 1.6278831 X5 - 0.7934611 X6 - 3.0566093 X7, n = 32, m = 7, r = 0.9968, ST = 15.72, SR = 1.4310, Ev = 99.59%, RMS = 1.239, F = 531.227 where the independent descriptor variables, X1-X7, refer to the elements, P1, P2, P3, P4, P5, P6, P7 in the molecular path vector for all samples in both FID and TCD training sets; n, r, ST, SR, Ev, RMS and F are the sample number, regression coefficient, total standard deviation, standard residual deviation, explained variance, rooted mean squared error and F-statistic value, respectively. To test both models by using back-propagation neural network (BPNN) with the topological structure NN(7-4-2) and the cross validation through leave-one-out (LOO) procedure, the correlation coefficient of cross validation is over 0.96. Because there exists a quite good linear relationship between the molar responses and molecular path parameters, BPNN (r = 0.989 and 0.968) does not show its nonlinear advantage over multiple linear regression(MLR) (r = 0.9976 and 0.9968) in both presently examined cases, FID and TCD in the GC technique, for molecular modelling and quantitative prediction. PMID- 12541732 TI - [Studies on retention behaviour of biopolymers on silica-based binding stationary phase in the mobile phase of acetic acid-water]. AB - The reversed-phase chromatographic method consisting of acetic acid-water as a new mobile phase for separating biopolymers on ODS column is presented. The new mobile phase may overcome the weak points of irreversible adsorption in chromatographic separation to some degree and difficulty in concentrating proteins with lyophylization and reduce the three-component (organic solvent, ion pairing agent and water) of typical mobile phase to two component system. Based on the retention and Z (the number of solvent molecules involving in the displacement from stationary-phase surface), log I (a constant relating to the affinity of 1 mol of solute to the bonded stationary phase), and j (a constant relating to the affinity of 1 mol of solvent to the bonded stationary phase), the elution strength of acetic acid-water mobile phase was found to be stronger than that of methanol-water-TFA. From a good linear relationship between log I and Z, it can be concluded that the interaction force between proteins and RPLC stationary phase with acetic acid-water as mobile phase is non-selective hydrophobic interaction forces. In addition, the molecular conformations of protein in acetic acid-water and methanol-trifluoroacetic acid-water were found to be different. PMID- 12541733 TI - [Determination of catechins in grape seeds by capillary electrophoresis]. AB - A capillary zone electrophoresis (CZE) method has been developed for the determination of four pharmacologically active catechins in grape seed. The components analyzed were (+)-catechin, (-)-epicatechin, epigallocatechin and epicatechin gallate. Separation was achieved by using a mixed buffer of 0.02 mol/L borate and 0.005 mol/L phosphate at pH 10.0 as background electrolyte and an uncoated fused silica capillary with 50 microns i.d. x 375 microns o.d. (effective length 41.5 cm) maintained at 25 degrees C. The applied voltage was 25 kV, and detection was performed by DAD at wavelength of 210 nm. By using proposed CZE method, four catechins were well separated within only 10 minutes with the repeatability of < 1% RSD for migration time and < 5% RSD for peak area. Correlation coefficients better than 0.995 were obtained within the concentration range from 0.005 g/L to 0.5 g/L for the four analytes studied with the detection limit of 3 mg/L-10 mg/L. The contents of these compounds in ten species of grape seeds cultivated in China were successfully determined. PMID- 12541734 TI - [Preparation and evaluation of polyvinyl pyrrolidone-coated columns based on sol gel chemistry for capillary electrophoresis]. AB - Polyvinyl pyrrolidone(PVP) coating techniques based on sol-gel chemistry was investigated in this article. Multiple-step coating and single-step coating methods were developed and the coating conditions(molar ratios of MAPS to 0.1 mol/L HCl, reaction times and concentration of PVP) were selected. Electroosmosis mobility and column efficiency were used as the evaluating parameters for coated capillary columns. For multiple-step coating, the optimal sublayer sol solution was obtained by adding 0.1 mol/L hydrochloric acid to MAPS(molar ratio, 1:1) and reacted at room temperature for 2 hours. The best PVP mass concentration was 40 g/L in alcohol. For single-step coating, the sol-gel solution was obtained by mixing the following ingredients: (a)MAPS, (b) 8% PVP alcohol solution and (c) 0.1 mol/L hydrochloric acid in 1:1 molar ratio to MAPS. The mixture was kept at room temperature for 2 hours. Proper amounts of 100 g/L ammonium persulfate and 10% (V/V) N, N, N', N'-tetramethylethylenediamine were then added. The mixture was mixed ultrasonically, then centrifuged at 4,000 r/min for 5 min. The upper clean sol-gel solution was transferred into another clean vial for coating capillary column. The reproducibility of migration times and efficiencies of single-step-coated capillary was better than that of multiple-step-coated capillary. Single-step-coating method was very effective and easy. It can fit the needs in experimental work. Coating techniques based on sol-gel chemistry are prospective but still need to be further developed. PMID- 12541735 TI - [Separation and determination of purine bases and pyrimidine bases from nucleic acid hydrolysis by HPLC on BDS column]. AB - The hydrolysates of nucleic acid, six purine bases and pyrimidine bases (cytosine, uracil, guanine, hypoxanthine, adenine and thymine) were separated and determined by using HPLC. It is discussed how the column and mobile phase affect the separation. The peaks of cytosine and adenine are tailed on ordinary C18 column, and they are very good on BDS-C18 column. The KH2PO4-H3PO4 buffer can be used in separation the hydrolysates of RNA and DNA, and the NaAc-HAc buffer is only used in DNA. In addition, pH value is a very important factor for separation. With pH value of mobile phase increasing, the retention times of guanine, hypoxanthine and thymine were first increased and then decreased, adenine was increased, and cytosine and uracil were almost constant. The chosen mobile phase was 0.1 mol/L KH2PO4-H3PO4 buffer, with a pH value of 4.05. It was detected at UV 260 nm. The determination was completed within 10 min. The RSDs were all less than 3% and the recoveries were in the range of 82%-114%. The method has been applied to the detection of yeast hydrolysates. PMID- 12541736 TI - [Advances of capillary electrophoresis and capillary electrochromatography using ionic coating column]. AB - Capillary electrochromatography (CEC) is a promising micro-separation technique, which combines the high selectivity of HPLC and the high efficiency of HPCE. In comparing to CE with fused silica capillary and CEC with ODS stationary phase, capillary electrophoresis and capillary electrochromatography with ionic coating columns can cause EOF more controllable, and provide an alternative separation mechanism. Therefore, the new CEC and CE modes are in favor of optimizing the separation and broadening the range of samples for analysis. The development of capillary electrophoresis and capillary electrochromatography using ionic coating column is reviewed with 48 references. PMID- 12541737 TI - Egg white protein-bonded columns and their applications. AB - Egg white protein-bonded columns were developed for HPLC. These columns can be used under aqueous mobile phase and separate various kinds of drug enantiomers. Hyphenated techniques using protein-bonded columns with LC/MS and/or column switching have been recognized as integral methods in pharmaceutical research to analyze drug enantiomers. Therefore, these methods are very useful for research fields of pharmacokinetics and pharmacology. PMID- 12541738 TI - [Determination of ferulic acid in chuanxiong and in animal serum and cerebrospinal fluid by reversed-phase high performance liquid chromatography]. AB - An easy, rapid and sensitive method for the determination of ferulic acid(FA) in Chuanxiong extracts, animal (mouse) serum and cerebrospinal fluid by RP-HPLC has been developed. The FA was separated on an ODS column, Nova-Pak C18(3.9 mm i.d. x 150 mm) and detected at the wavelength of 320 nm. The mobile phase was methanol water-acetic acid (35:65:0.5, V/V), with a flow rate of 0.8 mL/min. The detection limit of FA was 1.7 micrograms/L(S/N = 3) and the calibration curve was linear within the range of 0.85 mg/L-4.00 mg/L(r = 0.99904, n = 6). The mean recovery from animal serum and cerebrospinal was 95%-102%. PMID- 12541739 TI - [Analysis of ginsenosides by high performance liquid chromatography/mass spectrometry/mass spectrometry(LC/MS/MS)]. AB - Ginseng, one of the most popular medicinal herbs used in traditional Chinese medicines, has been studied to have biological effects attributing to its main constituents such as ginsenosides. There are more than 30 kinds of ginsenosides and some of them have similar polarity and structure. Thus, it is difficult to separate those ginsenosides with isocratic mobile phase owing to their similar polarity. A reversed-phase high performance liquid chromatographic method with gradient elution has been developed in this work. Nine ginsenosides can be separated by this method. Among them, five ginsenosides with different molecular weights can be determined by means of their molecular weight gained through LC/MS with an ESI interface. Re and Rd have the same Mw but belonging to two different kinds of ginsenosides. They can be differentiated from each other through their MS/MS gained by CID(collision induced decomposition) at the second stage quadrapole. Rc and Rb3 having the same molecular weight and belonging to the same kind of ginsenoside can be distinguished through their negative ion spectra gotten from LC/MS. So 9 ginsenosides can be determined by LC/MS/MS. This method can also be used to determine the ginsenosides contained in commercially available samples. PMID- 12541740 TI - [Simultaneous determination of sulbactam sodium and cefoperazone sodium in sulperazon by high performance liquid chromatography]. AB - A reversed-phase, isocratic high performance liquid chromatographic method with acid mobile phase can separate sulbactam and cafoperazone within 12 minutes. Column packed with Hypersil ODS2(250 mm x 4.6 mm i.d., 5 microns) was manufactured by Dalian Elite Company. Mobile phase is composed of water (adjusted to pH 4.0 with 1% phosphoric acid) and acetonitrile (80:20, V/V). The detection was performed at 210 nm and the injection volume was 2 microL. Cefoperazone and sulbactam have good linearity in the ranges of 100 mg/L to 800 mg/L and 100 mg/L to 1,000 mg/L with the correlation coefficients of 0.9991 and 0.9997 respectively. This method is easily to be operated and can be applied for manufacturing and medicinal study. PMID- 12541741 TI - [Determination of lactic acid in fermentation broth of Rhizopus oryzae by reversed-phase high performance liquid chromatography (RP-HPLC)]. AB - A method for determining lactic acid in fermentation broth of Rhizopus oryzae by RP-HPLC is described. The operating conditions were Wakosil-II 5 C18 RS column(4.6 mm i.d. x 150 mm, 5 microns) at room temperature, 0.01 mol/L phosphoric acid solution (pH 2.5) as mobile phase with a flow rate of 1.0 mL/min and UV detection at 210 nm. The retention time of lactic acid was 3.820 min. This method is simple, rapid and accurate. The results will not be affected by other components in the broth. The relative standard deviation was 0.22% (n = 5), and the recovery was over 99%. PMID- 12541742 TI - [The preliminary study on the open tubular capillary affinity liquid chromatography]. AB - A novel mode of affinity chromatography based on the open tubular column (OTAC) is presented in this paper. The OTAC column was prepared by coating a layer of Cibacron Blue F3GA onto the inner surface of capillary column with 50 microns inner diameter. The affinity chromatography experiment was performed on a capillary electrophoresis instrument by using its pressure system as driving force. The bovine serum albumin (BSA) and lysozyme (Lys) were successfully separated with step gradient elution. The relative standard deviation (RSD) for 10 consecutive runs for the elution time of lysozyme was less than 0.1%, and those for its peak area and peak height were 4.3% and 3.7% respectively. The good linear relationships of the amount of the injected lysozyme versus its peak area and peak height were observed with correlation coefficients of 0.9946 and 0.9988 respectively in the range from 8.6 ng to 28.7 ng. PMID- 12541743 TI - [A study on the mechanism of reductive alkylation for preparing 3-(beta-hydroxy ethyl-sulfonyl) N-ethyl aniline with HPLC/MS]. AB - Hydrogenating 3-(beta-hydroxy-ethyl-sulfonyl)-aniline and acetaldehyde in the presence of Raney Nickel as a catalyst, 3-(beta-hydroxy-ethyl-sulfonyl)-N-ethyl aniline was obtained with 98% conversion and 95% monoalkylation selectivity under optimum conditions. By using high performance liquid chromatography/mass selective detection technique to characterize the structures of the products, the mechanism of reductive alkylation is proposed. From the intermediates determined, it is shown that the reaction mechanism would go via an unstable N-alpha hydroxyethylaniline derivative and Schiff base stage. After hydrogenation of Schiff base, finally the product 3-(beta-hydroxyethyl-sulfonyl)-N-ethyl aniline was formed. PMID- 12541744 TI - [Enantiomeric separation of ketoprofen on an ovomucoid column]. AB - The enantiomeric separation of ketoprofen in three mobile phase systems on an ovomucoid (OVM) chiral stationary phase (CSP) column was investigated. Suitable chromatographic conditions for simultaneous separation of the two pairs of enantiomers of ketoprofen and its methyl ester were adopted. All of the four compounds occuring in the process of enzymatic stereoselective hydrolysis or esterification can be simultaneously separated. PMID- 12541745 TI - [Simultaneous rapid determination of eight food additives in foods by reversed phase high performance liquid chromatography]. AB - A simple and rapid method for the determination of eight food additives by RP HPLC is described. They were saccharin, aspartame, benzoic acid, sorbic acid, vanillin, caffeine, carmine and sunset yellow. The experiments were carried on Shim-pack CLC-ODS (150 mm x 6.0 mm i.d.) with methanol-20 mmol/L NH4Ac (44:56, V/V; pH 7.0) as the eluent at a flow rate of 1.0 mL/min. The UV detection wavelength was fixed at 220 nm. The food samples, after precipitation of the impurities with Carrez reagent, were injected directly into the HPLC system. The average recoveries of all the eight additives were between 91.9%-108.5%, and the RSDs were lower than 4% (n = 5). The analysis of a single sample required only 8 min. This method has been successfully applied to the routine analysis of these additives in foods. PMID- 12541746 TI - [Determination of bromhexine in plasma by gas chromatography-electron capture detection and pharmacokinetic studies]. AB - This paper reports a gas chromatography-electron capture detection(GC-ECD) method for the study of pharmacokinetics of bromhexine in healthy human body. A2 m x 3 mm i.d. silanized glass column packed with 5% SE-30 was used. The carrier gas was nitrogen. The internal standard was 5-chloro-2-amino-diphenyl ketone. After NaH2PO4-Na2HPO4 buffer (pH 6.0) being added, the plasma was extracted with n hexane-dichloromethane (9:1, V/V). A good linearity was obtained from 1.0 microgram/L to 50.0 micrograms/L of bromhexine in human plasma, r = 0.9994. The detection limit of bromhexine in plasma was 0.5 microgram/L. The average recovery was 97.5%. The pharmacokinetics of bromhexine was determined by this method after a single oral dose of 8 mg bromhexine capsule given to each of 8 volunteers. The results showed that the plasma concentration-time courses conformed to one compartment model. The established GC-ECD method is a good method for the determination of bromhexine in human plasma. The method is rapid, simple, precise and sensitive. PMID- 12541747 TI - [Practical optimization of the two-dimensional thin-layer chromatographic separation of ginsenosides]. AB - A computer-assisted method is presented for optimization of mobile phase selection for separation of the mixture of ginsenosides in two-dimensional TLC. This method bases on the correlation between the proportions of mobile phases and the retention values Rf of ginsenosides, and uses Dmin(the minimum distance of two spots) as the selection criterion with a two factor (Xs, Ys) statistical scanning technique. The experimental results, from which we observe that more new spots are separated than on one-dimensional TLC, are in excellent agreement with the predicted results. PMID- 12541748 TI - [Study of the detection characteristics of clozapine, N-desmethyl clozapine and olanzapine with high performance liquid chromatograph-electrochemical detector]. AB - In order to analyze clozapine, N-desmethyl clozapine and olanzapine, their detection characteristics with high performance liquid chromatograph electrochemical detector (HPLC-ECD) were investigated. The separation was performed on an ODS-3 column with the mobile phase of methanol and 0.1 mol/L phosphate buffer(60:40, V/V). The retention times of clozapine, N-desmethyl clozapine and olanzapine were all prolonged with higher pH of the mobile phase. These three compounds could be separated on the baseline at pH 4.56 and 5.56. The relationships of peak heights and detection voltages shown typical "S" shaped curves, and these curves shifted to the left with higher pH. To get stable detection current, the detection voltages for clozapine, N-desmethyl clozapine and olanzapine must be higher than 0.60 V, 0.60 V and 0.35 V at pH 4.56, and 0.48 V, 0.48 V and 0.30 V at pH 5.56, respectively. The typical "S" shaped ampere-volt curves were very important for the selection of suitable voltage for quantitative detection, and could be used for the qualitative detection of these three compounds. PMID- 12541749 TI - [Determination of Pericarpium papaveris in foods accompanied with some fragrant substances by high performance liquid chromatography]. AB - An HPLC method for the determination of Pericarpium papaveris in foods accompanied with some fragrant substances is described. The Pericarpium papaveris in food was extracted in alkaline condition with CHCl3-C2H5OH(9:1, V/V) and analyzed by reversed-phase HPLC by using a YWG C6H5 column and acetonitrile methanol-water as mobile phase. The calibration curves of papaverine, morphine and codeine were linear in the range of 1.2 mg/L-100 mg/L and the mean recoveries were from 79.3% to 86.1%. The method has a good specificity and accuracy. PMID- 12541750 TI - [Separation and determination of monosaccharides with high performance liquid chromatography by using internal standard method]. AB - A new approach by which free plant monosaccharides were separated directly and determined with high performance liquid chromatography by using internal standard method was established and beta-indolylacetic acid was selected as the internal standard substance. The separation time for xylose(Xyl), fructose(Fru), glucose(Glu), sucrose(Suc), maltose(Mal), lactose(Lac) and raffinose(Raf) was within 18 min; and the detection limits were 1.8 micrograms, 2.3 micrograms, 2.7 micrograms, 1.8 micrograms, 3.5 micrograms, 4.1 micrograms and 4.3 micrograms respectively while the linear dynamics varied within the range of 5.0 micrograms/L-750 micrograms/L. In this work, the effects of the concentration of CH3CN and pH values in the mobile phase to the separation of the 7 sugars and beta-indolylacetic acid were studied as well. The method can be used to the determinations of jujube, apple samples. Moreover, the recovery experiment was also conducted. The results showed that the relative standard deviations of 5 determinations to Xyl, Fru, Glu and Suc were 1.66%-3.45%, 1.54%-3.01%, 1.29% 2.91%, 1.09%-2.65% and the recoveries were 97.4%-102.1%, 97.3%-101.8%, 98.7% 102.2%, 97.7%-102.5%, respectively. PMID- 12541751 TI - [Liquid-solid adsorption separation and gas chromatography/mass spectrometry analysis of recycle alkanes in the production of alkyl benzenes]. AB - A method for qualitative and quantitative analysis of recycle alkanes in the production of alkyl benzenes is proposed in this paper. The mixture was separated into saturated hydrocarbons (alkanes and cyclanes) and aromatics by means of adsorption chromatography with silica gel of 100 mesh-200 mesh as stationary phase and n-pentane and dichloromethane as eluents. Results were obtained by analysing these two fractions separately. A total of 21 different kinds of n-, iso-, cyclo and a few light aromatic hydrocarbons were identified in the saturated hydrocarbons which were 97.3% of the mixture, while the rest 2.4% aromatic hydrocarbons included 15 different kinds of indanes, naphthanes and diphenylanes, the majority of which were products of dehydrogenation and alkylation, and the minor part was from original alkanes. By-products can be truely revealed by the qualitative analysis, which can be used to improve the process conditions. Analysis showed that after adsorption separation by silica gel, the low-content aromatic hydrocarbons was concentrated, and thus the analysis accuracy and sensitivity were improved. Preseparation avoided the problems which would appear if the samples were directly analysed by gas chromatography or gas chromatography/mass spectrometry. PMID- 12541752 TI - [Study on the analysis of alpha-naphthylacetic acid]. AB - This paper introduces a method for the determination of alpha-naphthylacetic acid with HPLC. Under the reversed-phase condition, puting ion-pair reagent IPR-A in the mobile phase of methanol-water, an unknown peak was separated. The recovery was 100.2% and the RSD was 0.30%. In comparing with conventional HPLC using methanol-water as mobile phase and GC methods, the results were more dependable. PMID- 12541753 TI - [Determination of benproperine phosphate tablets by high performance liquid chromatography]. AB - A reversed-phase HPLC method for the determination of benproperine phosphate tablets is reported. The chromatographic conditions were Hypersil-C18 column(5 microns, 4.6 mm i.d. x 150 mm) at 35 degrees C, mobile phase of methanol-water glacial acetic acid-triethylamine(60:35:5:0.1, V/V) with a flow rate of 0.9 mL/min and UV detection at 270 nm. Cortisone acetate was selected as the internal standard. The linear relationship of calibration curve was good in the range of 9.96 mg/L-49.8 mg/L(r = 0.9998). The average recovery and RSD were 99.91%(n = 5) and 0.43% respectively. The total time of analysis for a run was within 7 min. The method is simple, sensitive, rapid and accurate. PMID- 12541754 TI - [Determination of phthalic diesters in water by solid phase microextraction capillary gas chromatography]. AB - The solid phase microextraction (SPME) has the advantages of high efficiency, high sensitivity, simple operation and solvent free operation for analyzing environmental samples. An optical fiber coated with mixed stationary phase of polysiloxane and polymeric fullerene (in ratio of 4:1) was used to extract 5 phthalic diesters by HS-SPME-GC. The operating conditions of SPME such as the extraction temperature, ionic strength of solution, adsorption time and desorption time have been studied. It was compared with commercial polymer coating PDMS fiber through headspace SPME combined with GC. The results indicated that OV-1/PSO-C60 was better than PDMS in extracting high boiling point and semi volatile phthalic diesters. For phthalic diesters, the detection limits were in the range of 0.331 ng/L-12.5 micrograms/L, and relative standard deviations were below 12% except for dinonyl phthalate. PMID- 12541755 TI - [Determination of residual acetone in soybean phospholipids by headspace gas chromatography]. AB - Soybean phospholipids have many functions and alimentary actions. In our country, powder soybean phospholipids are generally got by extraction with acetone, followed by vacuum drying. There may be some residual acetone present in the soybean phospholipids, which is harmful to health. So, we must know residual acetone content in the soybean phospholipids. However we have not found a method to determine the residual acetone in the soybean phospholipids. In this paper, headspace GC was used to determine residual acetone in powder soybean phospholipids. The headspace bottle was glass with a volume of 15 milliliters. Certain amounts of water, ammonium sulfate, and sample were added into the bottle. The mixture was made into a brei as soon as possible. The bottle was put into a water bath at 40 degrees C for an hour. The GC column was a 2 m x 3 mm i.d. stainless steel tube packed with GDX-103 stationary phase. Temperatures of both injector and detector were kept at 120 degrees C. Column temperature was 160 degrees C. Injection volume was 1 mL. External standard method was used for quantitation. The RSD was 1.2%. The recoveries in the range of 25.0 micrograms/g 100 micrograms/g were 98.4%-104%. PMID- 12541756 TI - [Application of capillary chromatography in benzene boiling range analysis]. AB - The experimental data were analyzed by regression analytical method and the relation between the quantity of impurities and the boiling range of benzene was demonstrated. It was proved that when the quantity of hydrocarbon impurities was within certain limits, the quality of benzene fractions can be determined by analyzing the hydrocarbon impurities in it. So the distillation method for analyzing benzene boiling range may be replaced by capillary chromatography for analyzing the hydrocarbon impurities. The practical applications support the above suggestion. PMID- 12541757 TI - [Purification of follicle-stimulating hormone by radial-flow chromatography]. AB - By comparing the purification results of radial-flow chromatography with conventional chromatography, a new purification process of follicle-stimulating hormone (FSH) from crude sample of human menopausal gonadotrophin has been developed. The results showed that the specific activity, activity recovery, and purification factor of FSH purified by radial-flow chromatography were higher than those by conventional chromatography in large scale. The specific activity of FSH purified was 180 IU/mg protein, activity recovery 56%, purification factor 150. The purified product meets the quality requirement of the Standard of Chinese Pharmacopoeia. PMID- 12541758 TI - [Analysis of the reason that patients with chronic sinusitis having a negative CT manifestation]. AB - OBJECTIVE: To explore the reason why the CT manifestation of the patients with chronic nasosinusitis is negative. METHOD: To analyze the clinical and CT manifestation of the patients with chronic sinusitis having a negative CT manifestation. RESULT: Pathological changes were found in all 8 patients during the course of endoscopic sinus surgery, such as pus storing in the sinus cavity, mucous swelling polypoidly, small polyps formation and so on. CONCLUSION: The reasons that the patients with chronic sinusitis having a negative CT manifestation are, 1. CT scan can only give a static one-off image, 2. partial volume effect, 3. maybe result from the location of CT scan and the resolving power of tomograph, 4. maybe attribute to the pathological classify of chronic sinusitis. PMID- 12541759 TI - [Expression of TNF alpha and VCAM-1 in nasal polyps and relation with eosinophil infiltration]. AB - OBJECTIVE: To study the expression of TNF alpha and VCAM-1 in nasal polyps tissue and its significance. METHOD: Paraffin sections of nasal polyps and chronic rhinitis samples were studied with immunohistochemical technique. RESULT: Expression of TNF alpha and VCAM-1 were stranger in nasal polyps tissues than in controls. There was positive relationship between the expression of TNF alpha and that of VCAM-1(r = 0.833), and expression of VCAM-1 coincide with eosinophil infiltration(r = 0.746). CONCLUSION: It is suggested that TNF alpha may up regulate the expression of VCAM-1 in vessel endothelium, and prompt adhesion and migration of eosinophils, TNF alpha and VCAM-1 may play important role in the pathogenesis of nasal polyps. PMID- 12541760 TI - [Linkage of allergic rhinitis with HLA-DRB alleles polymorphism]. AB - OBJECTIVE: To investigate whether susceptibility or resistance to allergic rhinitis associated with HLA-DRB alleles. METHOD: Using PCR-SSP (sequence specific primer polymerase chain reaction), we tested the frequency distribution of HLA-DRB alleles in 41 patients with allergic rhinitis (AR) and 41 healthy controls from Beijing China. RESULT: The frequence of HLA-DRB1 * 0301.2 and HLA DRB4 * 0101 was lower in AR than in controls (2.44% vs 17.07%, P < 0.05; 29.27% vs 51.22%, P < 0.05). CONCLUSION: HLA-DRB1 * 0301.2 and HLA-DRB4 * 0101 alleles might confer protection against AR. PMID- 12541761 TI - [Clinical observation of TNF-alpha content in nasopharyngeal secretion of patients with nasopharyngeal carcinoma]. AB - OBJECTIVE: To detect the changes of tumor necrosis factor-alpha (TNF-alpha) in nasopharyngeal secretion of patients with nasopharyngeal carcinoma(NPC). METHOD: The content of TNF-alpha in nasopharyngeal secretion was determined by radioimmunoassay (RIA) in 20 controls, 52 patients with NPC. RESULT: The average concentrations of TNF-alpha were (12.95 +/- 4.62) pmol/L in control group, (33.68 +/- 15.13) pmol/L in NPC group and (64.00 +/- 11.57) pmol/L in NPC with necrosis group. The content of TNF-alpha in nasopharyngeal secretion in control group was lower significantly than that of patients with NPC (P < 0.01). CONCLUSION: The level of TNF-alpha in nasopharyngeal secretion of NPC patients was higher than that of normal subjects. To detect the content of TNF-alpha in nasopharyngeal secretion is a useful test for the study and diagnosis of NPC. PMID- 12541762 TI - [Measurement of serum neopterin levels in patients with nasopharyngeal carcinoma and its significance]. AB - OBJECTIVE: To evaluate the relationship between serum neopterin levels and nasopharyngeal carcinoma and its clinical significance. METHOD: The serum neopterin levels were determined in 54 patients with nasopharyngeal carcinoma (NPC group) and 32 healthy adults (normal group) by ELISA. RESULT: The serum neopterin levels in NPC group were significantly higher than that of in normal group (P < 0.001). The serum neopterin levels in UICC-TNM stages were undifferentiated (P > 0.05). The serum neopterin levels in patients with no lymph node metastasis were lower than that of in lymph node metastasis (P < 0.05). The serum neopterin levels decreased significantly after disappearance of tumor and its lymph node treated by radiotherapy (P < 0.01). CONCLUSION: Cellular immunity mediated by macrophages might be involved in the pathogenesis of nasopharyngeal carcinoma and the measurement of the serum neopterin may be a useful objective index to evaluate cellular immunological functions and curative effects. PMID- 12541763 TI - [Expression and significance of TGF-beta 1 in nasal polyps]. AB - OBJECTIVE: To study the expression and significance of transforming growth factor beta 1(TGF-beta 1) in nasal polyps. METHOD: Expression of TGF-beta 1 in nasal polyps from 34 patients and middle turbinates from 30 patients with deviation of nasal septum were prospectively studied with immunohistochemistry. Each tissue section was observed under optical microscope. RESULT: 1. The TGF-beta 1 positivity in extracellular matrix and in cells in the stroma was significantly higher in nasal polyps than in middle turbinates (P < 0.01). 2. The distribution and shape of TGF-beta 1 expressing cells in nasal polyps was similar to that of eosinophil, their positivities were significantly correlative (P < 0.05). 3. The positivity of TGF-beta 1 did not correlate with clinical type of nasal polyps (P > 0.05), eosinophil infiltration correlated significantly with clinical type of nasal polyps(P < 0.05). CONCLUSION: 1. The TGF-beta 1 may contribute to some of the pathologic changes observed in nasal polyps, such as thickening of the epithelial basement membrane and stromal fibrosis. 2. Eosinophils in nasal polyps represent a major source of TGF-beta 1. 3. Eosinophils infiltration may play a prominent role in the development and recurrence of nasal polyps. PMID- 12541764 TI - [The impact of lateral rhinotomy on nasal airway]. AB - OBJECTIVE: In order to ascertain the lateral operation and medial maxillectomy affect on nasal airway. METHOD: Report 16 cases of nasal cavity or nasal sinus neoplasm who were therapied by lateral rhinotomy and medial maxillectomy. They were observed by acoustic rhinometer (AR) and CT examination in 1-2.5 years after the surgery. RESULT: The feeling of nasal obstruct were disappeared in 3 months after the surgery. The nasal airway resistance(NAR) of the ill nasal cavity was markedly decreased after the surgery, and the total NAR were decreasing unmarkably. The mucous proliferation was notable only in the first year after surgery, and especially on remains of the inferior turbinate or extra wall of nasal cavity. CONCLUSION: Lateral rhinotomy operation and medial maxillectomy have less impact on nasal airway and the functions. PMID- 12541765 TI - [Human papillomavirus DNA test in nasal polypsis]. AB - OBJECTIVE: To evaluate pathogenesis of nasal polypsis. METHOD: To detect HPV in 26 pathological samples of 13 cases by polymerase chain reaction(PCR). Ten cases of normal nasal mucous were used as control. RESULT: HPV-DNA was positive 12 cases(92.3%) in 13 cases of the first surgical operation by PCR. HPV-DNA was positive 5 cases (38.5%) in 13 cases of the second surgical operation by PCR. The most HPV type was HPV6. HPV-DNA was negative in 10 cases of control. CONCLUSION: The results showed that HPV plays an etiologic role in the development of nasal polypsis. PMID- 12541766 TI - [The determination of p53 protein in maxillary sinus carcinoma and its prognosis value]. AB - OBJECTIVE: To determine whether p53 protein overexpression correlated with early tumor recurrence in carcinoma of maxillary sinus. METHOD: Forty-five patients with T2N0M0 squamous cell carcinoma of maxillary sinus were treated with operation and/or radiotherapy. The median follow up time was 3 years. The ratio of local control was 62.2% (28 patients), as control group; The ratio of local recurrence and/or metastasis was 37.8% (17 patients), as experiment group. Cancer specimens were obtained before treatment. p53 protein overexpression, which indicated a mutated p53 gene, was analyzed by immunohistochemistry, using the mouse monoclonal antibody DO-7. RESULT: Approximately 88.2% of the carcinoma that recurred locally overexpressed p53 protein, compared with only 28.6% of those with local control (P < 0.01). No significant relation was noted between p53 protein overexpression and pathohistologic grade. Intensity of staining did not predict tumor recurrence. CONCLUSION: The study demonstrated that the p53 protein is an important prognostic marker on gene level for maxillary sinus carcinoma. PMID- 12541767 TI - [Transconjunctival approach to the fractures of the orbital floor and infraorbital rim]. AB - OBJECTIVE: To study the surgical approach to the orbital floor and infraorbital rim. METHOD: 15 cases of fractures of the orbital floor and infraorbital rim were treated with the transconjunctival approach. RESULT: All fractures of the orbital floor and infraorbital rim were repositioned and fixed through transconjunctival approach. Except for 1 case with mild canthal malposition, no other complication was observed. CONCLUSION: The transconjunctival approach can provide adequate exposure of the infraorbital rim and orbital floor without scar formation in the face. PMID- 12541768 TI - [Influence of anisodamine on guinea pig experimental nasal hypersensitivity]. AB - OBJECTIVE: To observe the influence of anisodamine on guinea pig experimental nasal hypersensitivity. METHOD: Experimental animals were divided into normal control group, TDI group, reserpine group and anisodamine group. The scores of symptom of nose and the cytologic changes of nasal mucosa were parameters of observing. RESULT: The nasal symptom of reserpine group was significantly stronger than the TDI group, and the eosinophilia evidently increased (both P < 0.01). The nasal symptom of the anisodamine group reduced and the eosinophilia decreased. It is significantly difference between the TDI group and the reserpine group (both P < 0.01). CONCLUSION: Anisodamine may play a role in therapy of the experimental nasal hypersensitivity with hyperfunction of parasympathetic nerve. PMID- 12541769 TI - [Functional subdivisions of the guinea pig accessory olfactory bulb revealed by electrophysiology]. AB - OBJECTIVE: To elucidate possible functional subdivisions in the guinea pig accessory olfactory bulb. METHOD: The guinea pig accessory olfactory bulbs were cut in sagittal slice. Bipolar tungsten electrodes were inserted into anterior or posterior vomeronasal nerve layers and single square-pulses were delivered through the electrodes to activate afferent fibres. Glass microelectrodes were used to record extracellular field potentials of anterior or posterior external plexiform layers. RESULT: A single shook of the VNL provoked a characteristic damped oscillatory field potential and the oscillation in the pAOB was more distinct in wave form and longer in duration than those in the aAOB. The stimulation of anterior VNL elicited field potentials exelusively in the anterior region of EPL, whereas shocks to the posterior VNL provoked oscillatory responses only within the posterior EPL. CONCLUSION: The accessory olfactory bulb in the guinea-pig is distinctly segregated into the anterior and posterior subdivisions and an anatomical boundary exists in both regions. The results suggested that there are at least two different input-output pathways in vomeronasal systems. PMID- 12541770 TI - [Recognizing of the transglottic carcinoma]. AB - OBJECTIVE: A study was undergone about transglottic carcinoma(TGC) in eighty six cases from 1979 to 1998, so as to recognize the TGC. METHOD: Evaluating retrospectively the procedure of the disease, surgical operations, presence of cervical metastasis, gathering the data on histopathological discovery and following up and so on. RESULT: The TGC was different from T2, T3 supraglottic or glottic carcinoma of larynx. In 86 TGC, there were 63 cases which were operated with total laryngectomy, 9 cases subtotal laryngectomy and 14 cases frontolateral hemilaryngectomy, 47 cases with neck dissection. In TGC which size > 2 cm, there were 58% which extend to the framework of larynx, 46% with extension out of larynx, and 71% with neck metastasis. The result of following up shows that 5 year survival rate was 55.41%, 10-year was 29.72%, 15-year and more was 18.92%. CONCLUSION: There is an actual signification for TGC to be a special type of the laryngeal carcinoma. The characteristics of the precise topography of the the paraglottic space (PGS) can explain the reason of extension, spread of the TGC and cervical metastasis. Elective neck dissections and complemental radiotherapy postoperatively should be included during surgical resection. PMID- 12541771 TI - [Study of the reconstruction methods after vertical partial laryngectomy for glottic carcinoma of the larynx]. AB - OBJECTIVE: To study the reconstruction methods after vertical partial laryngectomy for glottic carcinoma of the larynx. METHOD: 58 cases with glottic carcinoma of the larynx were treated with vertical partial laryngectomy or extended vertical partial laryngectomy. The clinical materials of these cases were retrospectively studied. RESULT: The total decannulation rate was 93.1% and the 3- and 5-year survival rates were 87.2% and 80.5% respectively. The voice was socially acceptable in 93.1%(54/58) of the patients. Aspiration was only 3. All resumed mouth-food-taking. CONCLUSION: The reconstruction methods after vertical partial laryngectomy were mainly decided by the extent of the laryngeal defect. If the laryngeal defect of the framework was small, the pedicle muscular flap was the best reconstruction method in comparison with other reconstruction methods. If the laryngeal defect of the framework was too big, allograft nasal septal cartilage or epiglottic laryngoplasty should be performed. PMID- 12541772 TI - [Clinical analysis of 62 cases laryngeal papilloma in children]. AB - OBJECTIVE: To investigate the clinical features of laryngeal papilloma in children. METHOD: In a group of 62 patients with laryngeal papilloma in children, the tumors of 28 patients were cut under the direct laryngoscope, 34 patients were treated with laryngeal micro-laser operation. RESULT: The post-operation followup ranged from 2 to 5 years, the cure rate of 2 years was 51.61%. CONCLUSION: The clinical features of this disease include rapid development, a large lesion, and it is often found in the infraglottic cavity. Furthermore, we believe that in order to eliminate the tumors more accurately and decrease recurrence, micro-laser surgery and the use of interferon is very necessary. PMID- 12541773 TI - [Correlation of p16 mutation and biological behavior in Chinese laryngeal cancer]. AB - OBJECTIVE: To investigate the correlation between p16 gene mutation and laryngeal cancer biological behavior as well as its prognosis in laryngeal cancer. METHOD: 24 specimens of primary laryngeal cancer and 10 speciments with benign lesion in larynx were examined for mutations in exon2 of p16 by using PCR-SSCP silver stainning technique. RESULT: Mutations frequency of laryngeal cancer was 62.5% (15/24). Nothing was found in 10 cases with laryngeal benign lesion. CONCLUSION: There is a strong correlation between p16 gene mutation and the biological behavior of chinese laryngeal cancer, such as histologic differentiation, invasion stage, and regional lymph nodes metastasis (P < or = 0.05). PCR-SSCP silver stainning technique is one of the most sensitive and simplest measure for detecting genetic mutation. It is worth using in clinical laboratory because of its readiness, repetition and lower cost. PMID- 12541774 TI - [The dynamic study of cytokines pre- and postoperation in patients with carcinoma of larynx]. AB - OBJECTIVE: To observe the dynamic changes of cytokines pre- and postoperation in patients with carcinoma of the larynx and provide reference for recurrence monitor and prognosis evaluation in postoperative patients with head and neck tumor. METHOD: Thirty-four patients with head and neck squamous cell carcinoma were included in this study. Serum levels of IL-2, sIL-2R and TNF-alpha were measured before surgery, 7 days, 30 days and 90 days after surgery with ELISA and RIA. RESULT: The serum levels of sIL-2R and TNF-alpha before surgery in patients with carcinoma of the larynx were significantly elevated compared with controls(P < 0.01), while the level of IL-2 was not different. Serum concentrations of sIL 2R and TNF-alpha in patients without recurrence found at 7 days after operation were slightly higher than those before operation, but there was not significant difference. Moreover, the levels of sIL-2R and TNF-alpha at 30 days and 90 days postoperation decreased gradually, especially the sIL-2R levels decreased more significantly. No statistical difference were found among serum IL-2 levels in non-recurrence patients. The serum levels of sIL-2R and TNF-alpha in patients with recurrence at 30 days after operation were lower than those before operation and increased at 90 days after operation, while the levels of IL-2 decreased gradually. CONCLUSION: Dynamic study of the cytokines pre and postoperation play reference role in recurrence monitor and prognosis evaluation after the larynx carcinoma were removed. PMID- 12541775 TI - [Study on the coagulation state of laryngeal cancer patients before and after operation]. AB - OBJECTIVE: To explore the coagulation state of laryngeal cancer patients before and after operation. METHOD: The eight parameters of coagulant system of 46 laryngeal cancer patients were examined before operation, 1 day and 10 days after operation, and compared with control group. RESULT: The result suggests, anticoagulant activity decreased and fibrinolytic activity increased in laryngeal cancer patients before operation. Laryngectomy trauma may worsen the abnormality. With the incision of tumors and recovery from operation trauma, anticoagulant activity and fibrinolytic activity of the patients gradually returned to normal level. CONCLUSION: Coagulation disorders in laryngeal cancer patients had existed in a way. With operation trauma, it may worsen the abnormality. So anticoagulant and fibrinolytic systems of these patients should be taken seriously to prevent thrombosis and bleeding. PMID- 12541776 TI - [Surgical treatment and reconstruction for the patients with advanced-stage tonsillar cancer]. AB - OBJECTIVE: To investigate the operative approach and reconstruction for the patients with advanced-stage tonsillar cancer. METHOD: 7 cases with advanced stage tonsillar cancer were operated from Dec. 1994 to May 1999. Among them, 5 cases were operated through combined neck-mandible-oral cavity approach (CNMOCA) and with immediate reconstruction. Forearm free skin (FFS) flaps were applied in 3 cases. Pectoralis major myocutaneous (PMM) flaps were applied in 2 cases. 2 other cases were operated separately through approach of oral cavity or lateral pharynx. 6 cases were operated with radical neck dissection. RESULT: 5 cases who were operated through CNMOCA or with reconstruction were followed up from 1 year to 5 years and 3 months. They had not recurrence of tumor and complication. All had function recovery of speech. Swallowing or appearance is satisfactory in the 3 cases with FFS flaps. 2 Other cases died as recurrence of tumor in 1 year and 2 years and 10 months post-operation. CONCLUSION: CNMOCA is able to provide a good exposure method to remove the extensive tumors completely for the patients with advanced stage tonsillar cancer. It is necessary to reconstruct immediately for the patients. PMID- 12541777 TI - [Diagnosis and restitution of nasal bone fractures in emergency management]. AB - OBJECTIVE: To improve the diagnosis of nasal bone fractures in emergency. METHOD: 161 cases underwent fracture taxis, 118 cases underwent taxis and septum corrected operation, 13 cases underwent taxis and nasal sinus operation. RESULT: 279 cases were cured and 18 cases were improved. CONCLUSION: Patients must be checked carefully also with CT scan, one must pay attention to the symptoms of septum fractures, and the possibility of craniocerebral injury and nasal sinus fractures. Severe injuries must be treated according to the emergency treatment principle; patients with septum fractures may be managed simultaneously with septum corrected operation. PMID- 12541778 TI - [Different sampling method affects the voice assessment results for the patient with vocal polyp]. AB - OBJECTIVE: In order to definitude the influence caused by the different sampling in voice assessment. METHOD: We comparing the results acquired by total section and subsection sampling. RESULT: The results acquired by subsection tended to normal more than those acquired by total section. CONCLUSION: Subsection sampling voice assessment might conceal the degree of the disease state of patients. PMID- 12541779 TI - [Uvulopalatopharyngoplasty for the treatment of obstructive sleep apnea syndrome in 70 cases]. AB - OBJECTIVE: To elevate the effect of uvulopalatopharyngoplasty (UPPP) and minimize the postoperative complications. METHOD: The data of 70 patients with obstructive sleep apnea syndrome diagnosed with polysomnography were analyzed. RESULT: 15 cases (21%) reported unchanged snoring and apnea and hyponea index. 55 cases improved after UPPP for at least 6 months. The complications included hypertensive crisis (2 cases), respiratory embarrassment (4 cases) in period of paretic UPPP. After UPPP, The complications included local hemorrhage (1 case), wound dehiscence (2 cases), temporal velopalatal insufficiency (9 cases), one had suffocation after extraction of endotracheal tube. CONCLUSION: Our results indicate that the strategies of improving the effect and avoiding the complications include: Excision of the lateral pharyngeal wall in retropalatal region and fat tissue in soft palate. Protection of palatosalpingeus and palatostaphylinus. Part preservation of uvula. Confirmation of the obstructive region. Choice of the anesthesia and preventive tracheotomy. Modified tracheotomy were safe and convenient when exchanging trachea canula. PMID- 12541780 TI - [Observation on the kinetic process in replantation of the recurrent laryngeal nerve of the dog]. AB - OBJECTIVE: In order to provide experimental foundation for replantation in the recurrent laryngeal nerve(RLM) of the laryngeal reconstruction and laryngeal transplantation. METHOD: The left RLN of ten caine dogs were cut. The histopathology of the cut RLN were observed under the microscope after operation (two dogs) and 3.5 hours later (two dogs). Meanwhile the RLN were sewed up under the microscope at once. The kinetic process in regeneration of the RLN were observeded 1 week (two dogs), 2 week (two dogs) and 4 week (two dogs) after the replantation. RESULT: The replantation of RLN of six dogs succeeded. CONCLUSION: The results confirmed that sewing up the cut RLN at once is helpful to get normal regeneration in the replantation. PMID- 12541782 TI - [The study of human papillomavirus(HPV) DNA expression in middle-ear carcinomas]. AB - OBJECTIVE: To study the expressions of human papillomavirus DNA in the middle-ear carcinomas. METHOD: A PCR method using consensus primers for the detection of HPV6,11,16,18 was applied in 5 biopsies of middle-ear carcinomas. Eight biopsies of mucosa in middle-ear and mastoid were concluded as controls. RESULT: It was found that a resulting 80% (4/5) of the middle-ear carcinomas contained high-risk type HPV DNA, while the 0%(0/8) of the controls. CONCLUSION: The high prevalence of high-risk-type HPV in carcinomas of the middle ear suggests that viral infections may be an important etiologic component in the carcinogenic process. PMID- 12541781 TI - [Role of epithelial cell proliferation and apoptosis in human middle ear cholesteatoma]. AB - OBJECTIVE: To determine the role of middle ear epithelial cell proliferation and apoptosis in the pathogenesis of human cholesteatoma. METHOD: 33 cholesteatoma, 25 auditory meatal skin of cholesteatoma and 10 normal human auditory meatal skin were detected by using immunohistochemistry analysis for cell proliferation with Ki 67, PCNA and PCNA antibody and TUNTEL methods for cell apoptosis. RESULT: PCNA expression and apoptosis cells exists different quantity in cholesteatoma, auditory metal skin of cholesteatoma and normal human auditory meatal skin and distributes in various cell layer. There were positive correlation between p53 and PCNA as well as negative correlation between p53 and apoptosis. CONCLUSION: The dyregulation of proliferation and apoptosis in keratinocyte is important in pathogenesis of cholesteatoma. PMID- 12541783 TI - [Synchronal ossicular chain reconstruct of chronic suppurative otitis media surgery (389 cases analyze)]. AB - OBJECTIVE: To discuss the best material and operation style of synchronal ossicular chain reconstruct for chronic suppurative otitis media patients. METHOD: Review and analyze the effect of ossicular chain reconstruct in 389 cases with chronic suppurative otitis media after surgery. RESULT: The result of hearing improve of macro-tympanum III mould of ossicular chain reconstruct with artificial ossicle was better than other materials and operation types. The outcome of hearing improvement of incudostapedial joint reconstruct was better than the group of macrotympanum III mould of ossicular chain reconstruct with aritificial ossicle. CONCLUSION: The result of hearing improvement of macro tympanum III mould of ossicular chain reconstruct with aritificial ossicle was better than other material and operation types. The effect of aritificial ossicle better than self-ossicle in ossicular chain reconstruct surgery. PMID- 12541784 TI - [Labyrinthine fistula caused by chronic suppurative otitis media]. AB - OBJECTIVE: To study the clinical feature and surgical management of labyrinthine fistula caused by chronic suppurative otitis media (CSOM). METHOD: A retrospective study of 450 patients with CSOM who underwent radical mastoidectomy between the years 1992 to 1999 in Department of Otolaryngology, Xijing Hospital was conducted. RESULT: 20 patients (4.4%) with labyrinthine fistula caused by CSOM were found. The average history of ear drainage was 22 years, 18 patients (90%) had subjective hearing loss, and 13 patients (65%) had dizziness. The fistula test was positive in 9 patients (45%). 2 patients had anacusis in the affected ear. The preoperative pure tone average for bone conduction thresholds was less than 30 dB in 11 patients, between 31 and 50dB in 3 patients, and more than 51 dB in 4 patients. The ears with a positive fistula test had worse preoperative hearing than those with negative test (P < 0.01). The fistula was detected by CT in 2 of 9 patients. Cholesteatoma was present in 18 patients (90%) and granulation tissue was present in 2 patients (10%). The lateral semicircular canal was the site of labyrinthine fistula. The lesion at the site of fistula was completely removed in 17 patients and exteriorized in the remaining 3. CONCLUSION: There are no reliable methods at present for preoperative diagnosis of labyrinthine fistula, ultimately, the most reliable way to identify a fistula is during surgery. The method of manipulation of labyrinthine fistula is based on the concrete conditions of fistula and patient. PMID- 12541785 TI - [Diagnose of congenital malformation of inner ear by CT and MRI]. AB - OBJECTIVE: To inquire into diagnose of congenital malformation of inner ear by CT and MRI. METHOD: To report one case of CT and MRI image of congenital malformation of inner ear, and review relational documents. Character of CT and MRI of inner ear and membranous labyrinth by MR with 3D-CISS sequence were analysed. RESULT: The bone structure of inner ear was shown by CT, the memberanous labyrinth was shown by MRI. CONCLUSION: The mutual benefit of diagnose of congenital malformation of inner ear by CT and MRI. It is important for diagnose of congenital malformation of memberanous labyrinth by MR with 3D CISS sequence. PMID- 12541786 TI - [Use X-ray or CR to evaluate the operation result of the cochlear implant]. AB - OBJECTIVE: To evaluate the cochlear implant operation by the use of X-ray or CR. METHOD: There are 15 ears 15 cases which were completed the cochlear implant operation, after operation the implant electrode array was shown by the use of X ray or CR. RESULT: The shape and station of those were normal, average insered depth was 23.05 mm, come up to provide standard. CONCLUSION: It was an economic, convenient, objective and worthy method to evaluate the cochlear implant. PMID- 12541787 TI - [Research on the immunological cause of auricular pseudocyst]. AB - OBJECTIVE: To elucidate the relationship between the auricular pseudocyst and the immunological function of patients. METHOD: The cyst fluid and blood sample had been detected for contents of IgG, IgA, IgM and complement C3 by radial immunodiffusion, immunocomplex (IC) contents by polyethylene glycol turbidimetry in 55 cases, anti-nuclear antibody (ANA) by immuno-fluorescent technique in 23 cases, extractable nuclear antibody (ENA) by immunotransfer technique in 24 cases. The frozen sections of cyst wall tissue of 24 cases had been detected for immunocomplex by immunoenzyme histochemistric method. The reactions were observed separately between the auricle tissue of healthy white rat, human embryo and cyst fluid, serum of patient and normal serum. RESULT: Contents of IgG, IgA, IgM and C3 in the cyst fluid were lower than the serum significantly (P < 0.01). No IC, ANA and ENA could been found both in serum and cyst fluid, but reactive immunocomplex was discovered on the auricle cartilagous tissue of patient, immunocomplex also be observed on human embryo aurical tissue when it reacted with cyst fluid, but complex hadn't be found when the auricle tissue of white rat and human embryo reacted with serum of patient and normal people. CONCLUSION: The reason of auricular pseudocyst may be related with the local autoimmune status of patient. PMID- 12541788 TI - [Plasma autoantibodies to heat shock proteins in idiopathic sensorineural hearing loss]. AB - OBJECTIVE: To identify the specificity of autoantibodies to heat shock proteins in idiopathic sensorineural hearing loss (ISNHL) and to evaluate the existence of autoantibodies. METHOD: Plasma samples were obtained from patients with ISNHL (n = 41), systemic lupus erythematosus (n = 22), and normal controls (n = 30). Plasma autoantibodies to heat shock proteins (HSPs) were detected by Western blot using purified recombinant human HSP27, HSP60, HSP71 HSP90 alpha and HSP90 beta as antigen respectively. RESULT: In the ISNHL, the incidence of plasma autoantibodies to recombinant human HSP27, HSP60, HSP71, HSP90 alpha and HSP90 beta were 12.1%, 7.3%, 51.2%, 4.8% and 0% respectively. The incidence of autoantibody to HSP71 was significantly greater than that in systemic lupus erythematosus and in normal controls (P < 0.01). CONCLUSION: These results indicated that the autoantibody to HSP71 was main autoantibodies to HSPS in ISNHL and its presence might serve as a marker for an autoimmune etiology of these patients' hearing loss. PMID- 12541789 TI - [Mitochondrial mechanisms of CPT-induced apoptosis in nasopharyngeal carcinoma cells]. AB - OBJECTIVE: To understand whether CPT-induced apoptosis is related to the alteration of mitochondrial transmembrane potentials (delta psi m). METHOD: A low differentiation human nasopharyngeal carcinoma (NPC) cell line of CNE-2Z cells were used as in vitro model. After treatment with 2 mumol/L CPT for 24 h, the delta psi m was detected by the staining of rhodamine 123 (Rh123), while apoptosis was confirmed by sub-G1 cell content and the double staining of hoechst33258 and propidium iodide (PI). RESULT: CPT induced delta psi m collapse and apoptosis. With 2 mumol/L CPT treatment for 24 h, delta psi mlow cell was increased to (19.0 +/- 3.0)%, while in the DMSO control, delta psi mlow was (7.0 +/- 1.4)% P < 0.05. CONCLUSION: The disruption of the delta psi m may be one of the mechanisms of CPT-induced apoptosis in CNE-2Z cells. PMID- 12541790 TI - [Eight cases of rhabdomyosarcoma in head and neck]. AB - OBJECTIVE: To improve the clinical diagnosis and treatment level of rhabdomyosarcoma in head and neck. METHOD: It was based on the documents that 8 cases of rhabdomyosarcoma in head and neck had been diagnosed by pathological diagnosis. RESULT: The 8 cases were treated by operation. Among 8 patients, 5 took radiotherapy after operation. 1 took chemotherapy after operation. We kept in touch with 7 patients, the survival rate of 5 years was 57.1% (4/7). CONCLUSION: We must pay attention to the painful or painless phyma in the otolaryngology and head and neck. It has the possibility of being rhabdomyosarcoma. We diagnose the rhabdomyosarcoma through biopsy and immuohistochemistry and with the references from the examination of CT, MR. In order to improve the survival rate of patients of rhabdomyosarcoma, we should use the comprehensive treatment, including operation, radiotherapy, and chemotherapy. PMID- 12541792 TI - [Audiological findings of the aging across the urban and rural of Suzhou]. AB - OBJECTIVE: To determine the epidemiological characteristics of old people's hearing. METHOD: Questionnaires, physical examination, audiometry and bio chemical tests were performed on the elders above 60 years old in part of the urban and rural area of Suzhou with random sample survey. RESULT: A total of 1,040 individuals was investigated, among which 505 were from urban, and 535 were from rural. 33 of 505 senior citizen (6.53%) were nososacusis, 282 (55.84%) were presbycusis and 21 (4.16%) were noise-induced deafness; In the rural area, 35 of 535 (6.54%) were nososacusis, 232 (43.36%) were presbycusis, and 4 (0.75%) were noise deafness. There was significant difference of the incidence of presbycusis between urban and rural. The audiometric thresholds chart manifested that the threshold elevated with age increasing especially in high-frequency. CONCLUSION: The etiology of hearing loss of elders was mainly due to presbycusis. The higher incidence of psychotic disorder in urban probably caused a correspondingly higher incidence of presbycusis. So the prevention and cure of some age-induced diseased (e.g. hypertension, arteriosclerosis and diabetes) may be helpful to release and improve presbycusis. PMID- 12541791 TI - [Distribution of beta-actin within hair cells in the chick: comparatived with those in the guinea pig cochleae]. AB - OBJECTIVE: To study the distribution of beta-actin in the chicken and guinea pig cochlea hair cells. METHOD: beta-actin immunoreactivity was observed by immunohistochemistry. RESULT: beta-actin was located in stereocilia of hair cells and cytoplasm of marginal epithelium cells which tectorial membrane attached in basilar papilla; In the corti's organ beta-actin was present in three row outer hair cells cytoplasm and the phalangeal process and cytoplasm of the tunnel cells. CONCLUSION: There is a common cytoskeletal protein beta-actin in the chick and guinea pig cochleae, but the distribution of beta-actin in two species are different significantly. PMID- 12541793 TI - [Uncharged monolithic capillary column modified with an anionic surfactant in electrochromatography]. AB - A monolithic capillary column was obtained by in-situ co-polymerization of butyl methacrylate with ethylene dimethacrylate. A mode of capillary electrochromatography (CEC) based on the dynamic adsorption of long chain sodium dodecyl sulfate(SDS) on the monolithic stationary phases was developed. Effects of SDS concentration and concentration of acetonitrile in mobile phase on the electroosmotic flow(EOF) of the column and the plate height on the electroosmotic velocity were investigated. Efficiency of monolithic capillary column with higher than 140,000 plates per meter for neutral aromatic compounds were obtained, and the relative standard deviations observed for dead time(t0) and retention times of neutral solutes were about 0.22% and less than 0.56% for ten consecutive runs, respectively. PMID- 12541794 TI - [Development of a gas chromatographic column system for the on-line analysis of trace tetrahydrofuran in hexane]. AB - A gas chromatographic system using home-made 7 microns thick cross-linked dimethylpolysiloxane columns for the on-line analysis of trace tetrahydrofuran in hexane in a rubber production facility was developed. The experimental parameters of the column system, including the temperature, flow rate as well as the back flushing and heart-cutting program were investigated. The total column-switching program was suggested. The system has been successfully operated for more than one year with good resolution, stability and precision (RSD < 5%) in analyzing trace tetrahydrofuran (0-250 x 10(-6), V/V) for industrial process control in the rubber production facility. PMID- 12541795 TI - [Separation of echinacoside by reversed-phase preparative high performance liquid chromatography]. AB - Echinacoside, one kind of phenylethanoid glycosides (PhGs), was isolated from the stems of Cistanche tubulosa (Schenk) R. Wight with a series of steps, including solvent extraction, D101 polymer adsorption column separation, Sephadex LH-20 separation, C18 column reversed-phase preparative high performance liquid chromatographic (RP-prep-HPLC) preparation and polyamide thin-layer chromatographic detection. The purity of the product was over 98%. The chemical structure of echinacoside was identified by 1H NMR and 13C NMR. To find out the optimum condition of mobile phase of RP-prep-HPLC, several systems were used in this work. Finally acetonitrile-1% HCOOH water (18:82, V/V) system was found to be the best. On the other hand D101 polymer adsorption column and Sephadex LH-20 were also effective for PhGs separation. PMID- 12541796 TI - [Purification of proteins in human plasma with new nylon affinity membranes]. AB - A series of affinity membranes were prepared by using microporous nylon membrane as matrix and activation method with s-triazine and 1,4-butanediol diglycidyl ether, separately. Three proteins with clinic value, gamma-globulin, plasminogen and thrombin, were purified from human plasma by one step with affinity membrane chromatography. The purities of gamma-globulin purified were higher than 83%, and purification folds were higher than 5. The purity of gamma-globulin obtained was higher than that of standard human gamma-globulin. The efficiencies of gamma globulin purification with affinity membranes prepared by two activation methods were equal. Plasminogen purified with affinity membranes prepared by s-triazine activation was higher than that by 1,4-butanediol diglycidyl ether activation. And SDS-PAGE analysis showed that the purities of gamma-globulin and plasminogen obtained were higher than that of the commercial product. In addition, the purification of thrombin from human plasma by one step with trypsin activation column and affinity membrane was studied. Thrombin with specific activity of 42 NIH unit/mg and 42.5 NIH unit/mg could be obtained from human plasma through affinity membranes prepared by s-triazine and 1,4-butanediol diglycidyl ether methods, respectively. PMID- 12541797 TI - Determination of ginkgolic acids from Ginkgo biloba leaves by reversed-phase argentation high performance liquid chromatography. AB - An analytical method has been firstly achieved for the quantification of ginkgolic acids from Ginkgo biloba leaves by reversed-phase argentation high performance liquid chromatography. Analytical sample was cleaned-up after addition of acidic salt solution and adsorbent to the matrix solution by counter extraction of analytes with hexane. Ginkgolic acids were determined by HPLC with methanol and 5% aqueous acetic acid (90:10, V/V) containing 0.03 mol.L-1 silver ion as mobile phase and UV detection at 310 nm. Results showed ginkgolic acids were separated successfully from each other and from other interfering components, which were confirmed by spectra analysis and purity assay. The linearity of the calibration curve was good in the range of 0.084 microgram-10.56 micrograms (r = 0.9998). The average recovery was 97.3% and RSD was 1.6%. The detection limit was 0.026 microgram (S/N = 3). The convenient method can be used as a reliable tool for the quantitative analysis of ginkgolic acids. PMID- 12541798 TI - [Advances in estimation of physico-chemical parameters of organic compounds by RP HPLC]. AB - Reversed-phase high performance liquid chromatography is currently used in the determination of some important physico-chemical properties of compounds, and its potential uses in areas of modeling environmental behaviour and pharmacological activity of compounds are desirable. This paper reviews the methodology, major areas, present situation and chief difficulties of RP-HPLC method for the estimation of physico-chemical parameters, and the perspective of this method. PMID- 12541799 TI - [Research on the separation of three isomers of hydroxybenzoic acid by capillary zone electrophoresis]. AB - The separation behavior of o-hydroxybenzoic acid, m-hydroxybenzoic acid and p hydroxybenzoic acid in high performance capillary zone electrophoresis was investigated using cetyltrimethylammonium bromide (CTAB) as reversed reagent for electroosmotic flow. The effects of pH of buffer solution and volume fraction of methanol on the separation, peak shape and elution order were studied. PMID- 12541800 TI - [Measurement of binding constants of metal complex by indirect ultraviolet affinity capillary electrophoresis]. AB - The binding constants of complexes formed by metal ions and ligands have been obtained by affinity capillary electrophoresis, according to the measurement of the migration times of metal ions effected by ligands in aqueous media, under the experimental model named electrophoretic peak shifting. This method has been applied to the research of magnesium ion with citric acid and manganese ion with tartaric acid. Under the condition of imidazole and acetate buffer (pH 5.01) at 20 kV running voltage, the migration times of metal ion with ligand of different concentrations have been measured. The estimated logarithmic values of binding constants after computation were 3.27 for magnesium-citric acid and 2.28 for manganese-tartaric acid, which showed that the results were in agreement with the data in previous literature. This method is applicable to the measurement of the binding constants of complex systems with 1:1 ratio. PMID- 12541801 TI - [Separation, purification and identification of angiotensin converting enzyme inhibitory silk fibroin peptide]. AB - Silk fibroin peptides could be obtained from soluble silk fibroin by enzymatic hydrolysis. Its hydrolyzates produced with Alcalase showed significant inhibitory activity against the angiotensin I-converting enzyme (ACE). One inhibitory peptide from the hydrolyzate at a degree of hydrolysis of 20% (sample A20) was purified and identified. Sample A20 was first isolated by size exclusion chromatography(SEC), eluted with 0.01 mol/L hydrochloric acid solution on a Sephadex G-15 column (1.6 cm i.d. x 100 cm). The peak of No. 5 on the SEC chromatography was further purified by reversed-phase HPLC (mu Bondapak C18 P/N 84176 column, 7.8 mm i.d. x 300 mm), eluted with a linear gradient elution with acetonitrile from 0% to 15% at temperature (30 +/- 2) degrees C. Then the pure peptide with ACE inhibitory activity was obtained, the amino acid sequence of which was identified as Gly-Tyr by mass spectrometry. PMID- 12541802 TI - [Analysis of trehalose in transgenic tobacco extracts by high performance liquid chromatography with evaporative light scattering detection]. AB - A high performance liquid chromatographic method with ethylenediamine dynamically modified silica column and evaporative light-scattering detection (ELSD) was developed for the direct determination of trehalose in transgenic tobacco extracts. The separation column (Zorbax Rx-SIL, 4.6 mm i.d. x 250 mm, 5 microns, Hewlett-Packard, USA) modified by ethylenediamine and a guard column (Zorbax Rx SIL, 4.6 mm i.d. x 12.5 mm, 5 microns) were used. The mobile phase was a mixture of acetonitrile-water(2.6:1, V/V) containing 0.03% ethylenediamine. Regression equations revealed the linear relationships (r = 0.996-0.999) between the concentrations and peak areas of carbohydrates detected by ELSD. The detection limits of ELSD (S/N = 3:1) were 10 mg/L, 20 mg/L, 10 mg/L and 10 mg/L for fructose, glucose, sucrose and trehalose, respectively. PMID- 12541803 TI - [Determination of catechol-O-methyltransferase activity in human erythrocytes by high performance liquid chromatography]. AB - A rapid assay method for catechol-O-methyltransferase (COMT) activity in human erythrocyts by high performance liquid chromatography (HPLC) with UV detection was established. Enzyme activity was determined from erythrocyte lysates using S adenosyl-L-methionine (SAM) as methyl donor and 3, 4-dihydroxybenzoic acid (DBA) as substrate. The 3-O-methylated reaction products were measured by HPLC with UV detection. The linear range of COMT was from 1 U/mL to 60 U/mL with an average RSD < 10%, and the detection limit was 0.5 U/mL(S/N > or = 5). PMID- 12541804 TI - [Separation of catechins and caffeine in tea polyphenol by isocratic elution high performance liquid chromatography]. AB - The catechins, particularly in green tea, have been found to possess anti mutagenic and anti-tumorigenic properties. As each catechin possesses distinct properties, a simple and rapid method that could be used for analysis of individual catechins in a complex mixture would be necessary. A relatively simple and rapid method for simultaneous separation of five catechins and caffeine in tea polyphenol by isocratic elution high performance liquid chromatography has been developed. The analysis could be finished within 30 min. They were measured using Resolve C18 column (at 43 degrees C) and UV detector (at 280 nm), water-85% phosphoric acid aqueous solution-acetonitrile-dimethyl formamide(DMF) (859:1:120:20, V/V) as mobile phase. There was a good linear relationship between the content of component and its peak area for catechins and caffeine, with the correlation coefficients of 0.9992-0.9999. The average recoveries (n = 5) were 83.33%-104.42%, and the relative standard deviations (n = 6) were 0.74%-1.43%. The effect of concentration of DMF in mobile phase was studied. PMID- 12541805 TI - [Simultaneous determination of chlorpheniramine, paracetamol and pseudo-ephedrine hydrochloride in composite tablets by high performance liquid chromatography]. AB - An HPLC method is described for the determination of chlorpheniramine, paracetamol and pseudoephedrine hydrochloride simultaneously in Nuonuo composite tablets. Chromatographic conditions were: column, YWG-C18 column (150 mm x 4.0 mm i.d., 10 microns); mobile phase, 25% methanol(0.05 mol/L sodium dihydrogen phosphate); flow rate, 1.0 mL/min; detection wavelength, 205 nm; injection volume, 20 microL. The detection limits of chlorpheniramine, paracetamol and pseudo-ephedrine hydrochloride were 1.16 mg/L, 0.15 mg/L and 1.82 mg/L respectively and the corresponding average recoveries were 98.35% (n = 5, RSD = 1.60%), 101.16% (n = 5, RSD = 1.50%) and 98.50% (n = 5, RSD = 1.59%) respectively. The proposed method is simple, rapid and accurate and has been satisfactorily used for the quantitative analysis of real samples. PMID- 12541806 TI - [Study of unified analytical method of compatibleness for traditional Chinese medicines. I. Separation, recognition of characteristic components and fingerprint analysis for single and complex medicines]. AB - It is difficult to separate, identify and quantify multi-component in complex traditional Chinese medicines (TCM). Many analytical methods for TCM have been established, but only a few components can be separated and quantified. For analyzing the multi-component and finding new method of compatibility analysis, a strategy--the multi-mode and multi-column chromatographic system and unified analytical method has been proposed by us. A part of the results from the study of astragalus root(A), Chinese angelica root(Ca.) and their complex prescription (Danggui Buxue Tang, DBT) with the analytical mode of reversed-phase HPLC/photodiode array detector as an example of the method for unified separation and recognition of characteristic components and fingerprint analysis was introduced. This is the first part of the results of compatibleness analysis for TCM. PMID- 12541807 TI - [Determination of syringin and rutin in 11 species of Saussurea by high performance liquid chromatography]. AB - Zorbax ODS column and gradient elution of acetonitrile-acetic acid solution as mobile phase were employed to determine the content of syringin and rutin in 11 species of Saussurea by RP-HPLC. The detection wavelength was 260 nm. The results showed that standard curves of syringin and rutin were linear in the range of 0.0525 microgram-1.05 micrograms and 0.0658 microgram-1.32 micrograms, respectively. The correlation coefficients were 0.9988 and 0.9999. The method provides new indices for quality control of Saussurea. PMID- 12541808 TI - [Study of diphacinone in biological samples by high performance liquid chromatography/diode array detector]. AB - An analytical approach has been developed for high performance liquid chromatographic determination of diphacinone extracted from liver, blood, urine and kidney of rabbit by solid phase extraction (SPE) cartridges (using SAX, CN or SILICA GEL) with coumarin as the internal standard. Diphacinone was separated by reversed-phase gradient chromatography with DAD detection at 286 nm. The Analytical column was Hypersil BDS C18(150 mm x 4.6 mm i.d., 5 microns) and the guard column was Phenomenex ODS(4 mm x 3.0 mm i.d.). The mobile phase was a gradient mixture of aqueous solution (A) and methanol solution (B) both containing 0.5% ion pair A. There was a good linear relationship between the concentration of diphacinone and the ratio of peak areas of diphacinone and coumarin (internal standard) (r = 0.9999). The linear range was 1 mg/L-100 mg/L, and the lower detection limit was 5 ng (S/N = 3). The average recoveries of diphacinone in urine, blood and liver were 88.4% (n = 3, RSD = 1.25%, SPE by CN column), 82.2% (n = 3, RSD = 1.67%, SPE by SAX column), 91.0% (n = 3, RSD = 2.77%, SPE by SILICA GEL column), respectively. PMID- 12541809 TI - [High performance liquid chromatographic determination of domoic acid in shellfish samples]. AB - Domoic acid in shellfish samples was determined by HPLC with DAD/UV detector at 242 nm. Samples were extracted with methanol-water followed by clean-up of the extracts with strong anion exchange solid phase extraction cartridge(3 mL LC SAX). Zorbax SB-C18 column, 150 mm x 4.6 mm i.d., and mobile phase of acetonitrile-0.1% aqueous trifluoroacetic acid(13:87, V/V) were used for the assay. The quantitative analysis was performed with external standard. The calibration curve of domoic acid was linear in the range of 1.0 m/L-25.0 mg/L and the detection limit was ca. 0.2 microgram/g. PMID- 12541810 TI - [Determination of phosphocreatine in muscular tissues by high performance liquid chromatography]. AB - The quantitation of energy stores(phosphocreatine, PCr) is of paramount importance in the study of living-tissues metabolism. This is more so in the heart, which depends to a very great extent on an uninterrupted aerobic metabolism to maintain its normal function. Availability of data on myocardial energy stores is, therefore, indispensable for assessing the responses of heart to drugs or stresses such as ischemia and hypoxemia. A simple and rapid method for the determination of phosphocreatine in muscular tissues by RP-HPLC has been developed. The chromatographic conditions were as follows: Zorbax XDB-C18 column(150 mm x 4.6 mm i.d., 5 microns), V(20 mmol/L KH2PO4 and 2 mmol/L tetrabutyl ammonium phosphate): V(acetonitrile) = 96:4(pH 5.8) mobile phase and UV detector at 215 nm. PCr in muscular tissues was extracted with 0.4 mol/L HClO4. The calibration curve showed a good linearity in 5 mg/L-100 mg/L(r = 0.9992). The average recovery was 99.34%. The limit of detection was 2 mg/L. The verified results demonstrated that this method is precise, accurate and can be used for determination of PCr in muscular tissue. PMID- 12541811 TI - [Enantiomeric separations of 1-1,2,4-triazole compounds by micro-high performance liquid chromatography]. AB - The enantiomeric separations of sixteen novel 1-1,2,4-triazole chiral compounds were investigated by micro-HPLC with a 200 mm x 100 microns i.d. capillary chiral OD column. The detection was performed at 223 nm. The mobile phase was a mixture of iso-propyl alcohol and n-hexane(20:80, V/V) at a flow rate of 30 microL/min and column head pressure of 6.9 MPa. Some of the enantiomers could be separated on baseline. In comparing with the separation results by the conventional HPLC method, the micro-HPLC method had the advantages of high-speed, economy, environmental friendliness etc. The effect of solute structure on enantio selectivity was investigated and the chiral recognition mechanism was also discussed. It was found that the main separation mechanisms involve the pi-pi association and inclusion between the substituent R and the spiral pocket of OD CSP. PMID- 12541812 TI - [A new method for predicting retention time of solutes under linear gradient elution conditions in RP-HPLC]. AB - A new method for predicting the retention time of solutes under linear gradient elution has been established. In this method, the migration distance of solutes in column and the mobile phase gradient were modified simultaneously according to the distribution of mobile phase in column. The prediction results of retention time of both weakly and strongly retentive solutes were accurate in different instrumental systems using this method. In order to test and verify this method, the retention time of fifteen amino acids and eight homologous compounds of benzene were predicted. The predicted results were compared with those predicted by the method in common use. The relative average errors between predicted and experimental values were 3.70% and 4.90% for the weakly retentive solutes, far smaller than those obtained by the reference method (23.61% and 31.16%). For the strongly retentive solutes, the relative average errors were 0.21% and 6.01% respectively, slightly smaller than those obtained by the reference method (0.81% and 6.69%). It is seen that this new method has many advantages. PMID- 12541813 TI - [Study on reversed-phase high performance liquid chromatography separation condition and determination method of organic acids]. AB - The pH values of the mobile phase which affect the chromatographic behavior of organic acids in RP-HPLC have been studied. It was considered that the problem of organic acids on an RP-HPLC column was dissociation. The equation for finding the optimum pH of mobile phase was pHlimit of colunm < or = pH < or = pKa -2. In addition the effects of contents of methanol and acetonitrile in mobile phase were studied. By using the formula the mobile phases used for the analysis of organic monobasic acids--formic, acetic, propionic and butyric acid and of organic dibasic acids--propane diacid, butane diacid, pentane diacid and hexane diacid were proposed. For the analysis of the monobasic acids, the wavelength of UV-detector was set at 210 nm and the mobile phase was a mixture of phosphate buffer solution(15 mmol/L, pH 2.1)-methanol(85:15, V/V) and at a rate of 1.5 mL/min and the column was Diamond C18, 150 mm x 4.6 mm i.d.. The average coefficient of correlation was 0.9997 and RSD was less than 0.90%. The mobile phase for organic dibasic acids was a mixture of phosphate buffer solution (20 mmol/L, pH 2.1)-methanol(75:25, V/V). The other conditions were the same as for organic monobasic acids. The average coefficient of correlation was 0.9998 and RSD was less than 0.70%. PMID- 12541814 TI - [Research on a new side-chain liquid crystalline polysiloxane as stationary phase for capillary column gas chromatography]. AB - A new stationary phase, side-chain liquid crystalline polysiloxane, was obtained and used in capillary column gas chromatography. Two kinds of side-chain polysiloxane, PSI5 and PSI6, were characterized by polarized light microscopy (POM), differential scanning calorimetry (DSC) and X-ray diffraction and proved to be smectic phase Sc. Two fused-silica capillary columns (0.25 mm i.d.) were coated with 5 g/L solutions of PSI5 and PSI6 in dichloromethane by the static method, named column 15 and column 16. Numbers of theoretical plate were 1,598/m 2,423/m (I5) and 3,191/m-4,235/m (I6) for anthracene at 200 degrees C-240 degrees C. These columns showed wide operation temperature range (I5: 50 degrees C-200 degrees C; I6: 100 degrees C-260 degrees C), outstanding thermostability and weak polarity. These columns exhibited unique selectivity for many organic compounds, especially positional isomers. For example, the mixtures of anthracene, phenanthrene and fluorene were well separated on column I5 and I6; dimethyl benzene isomers were well separated on column I5, but they were not separated on column I6; dimethyl phenol isomers were well separated on column I5, but they were not wholly separated on column I6. The different selectivities of these columns are mainly due to the different chemical structures. PMID- 12541815 TI - [Structure characterization of melanin in black sesame by GC/MS]. AB - Black sesame melanin, a kind of biopolymer was degraded by alkali fusion to study structure characterization. The degraded products were derivatized with bis (trimethylsilyl) trifloroacetamide in a sealed tube at 125 degrees C for 30 min. The silylanization derivatives of degradation products were analyzed by GC/MS. Catechol, 1,4-dihydroxy benzene and catechuic acid were detected. This method can be used to characterize the structure type of black sesame melanin. PMID- 12541816 TI - [Determination of fenfluramine hydrochloride in tablets by gas chromatography with derivatization]. AB - The contents of fenfluramine hydrochloride in tablets were determined by using gas chromatography (GC) with derivatization. Five percent SE-30 was used as stationary liquid(2.1 m x 3.2 mm i.d., glass column), and was detected with flame ionization detector (FID). The oven temperature was set at 125 degrees C, with injector and FID temperature at 160 degrees C. The fenfluramine hydrochloride in the tablets was extracted with ethyl acetate and derivatized with trifluoroacetic anhydride at 40 degrees C for 30 min, and then analyzed by GC. The quantitative determination was performed with mexiletine as the internal standard. The method showed good linearity within the mass concentration range from 0.1 g/L-0.5 g/L(r = 0.9996) and the limit of detection was 8 ng. The average recovery of the samples was (100.2 +/- 2.2)% (n = 6). The precision of the method is satisfactory(RSD = 1.4%). The method developed has been applied to the determination of fenfluramine hydrochloride in tablets. PMID- 12541817 TI - [Determination of organophosphorus pesticide multi-residues in tea by gas chromatography]. AB - An efficient gas chromatographic method for the determination of 14 organophosphorus pesticide residues in tea by gas chromatography using micro chemical and solid phase extraction technique has been established. The organophosphorus pesticides were extracted from tea samples with ethyl acetate and mixture of ethyl acetate-hexane(1:1, V/V). The extracts were cleaned up with a column of 3 mL Supelclean Envl-carb by GILSON ASPEC XL. The 14 organophosphorus pesticides were separated from each other through SPB-1701 capillary column(30 m x 0.53 mm i.d. x 1.0 micron) and determined by flame photometric detector. The recoveries ranged 70.8%-120% when the added levels were from 0.100 mg/kg to 1.00 mg/kg, the RSD was 2%-7% and the limits of detection were 0.01 mg/kg-0.08 mg/kg. This method is rapid, sensitive and suitable for the analysis of pesticide residues. It can also be applied to the determination of organophosphorus pesticide multi-residues in other plant samples such as vegetables, fruits and so on. PMID- 12541818 TI - [Simultaneous determination of 11 organic acids in fruit juice by ion exclusion chromatography]. AB - A method for the quantitative determination of 11 organic acids (oxalic acid, citric acid, tartaric acid, malic acid, ascorbic acid, lactic acid, succinic acid, formic acid, acetic acid, glutaric acid and fumaric acid) in fruit juice was developed successfully. It was based on an ion exclusion chromatographic separation under the conditions of isocratic elution with 17 mmol/L sulphuric acid solution with the ICE-ION-300 ion exclusion column, and the UV detection at 210 nm. The precision of the method was investigated and the relative standard deviations were from 1.5% to 9.8% (n = 10). PMID- 12541819 TI - [Determination of bifonazole in cream by high performance liquid chromatography]. AB - An accurate method for the determination of bifonazole in cream is described. The analytical column was packed with Hypersil ODS. The mobile phase was a mixture of 0.08 mol/L aqueous triethylamine (TEA) phosphate solution(pH 7.0)-acetonitrile methanol (20:10:70, V/V) and the detection wavelength was 254 nm. This method is simple, accurate and can be used for the quality control of bifonazole in cream. PMID- 12541820 TI - [Analysis of olanzapine in human plasma with reversed-phase high performance liquid chromatography]. AB - Olanzapine is a novel antipsychotic drug. Recently it has been applied to clinical research. For the determination of olanzapine in plasma a reversed-phase HPLC method has been established. An aqueous solution of 0.25% ascorbic acid was added into the plasma sample and kept at -20 degrees C. Olanzapine can be separated on a Zorbax ODS column with a mobile phase of 50 mmol/L sodium phosphate (pH 7.2)-methanol-acetonitrile(12:10:3, volume ratios) and detected at 270 nm. The flow rate was 1.0 mL/min. The linear range was 15 micrograms/L-1,200 micrograms/L, r = 0.9988. The limited concentration of detection was 3 micrograms/L. The average recovery was (97.02 +/- 3.11)%. The intra-day and inter day coefficients of variation were 3.86% (n = 15) and 4.12% (n = 15) respectively. The method has been used to determine olanzapine concentration in patient's plasma. The data obtained show that the sensitivity and selectivity of this method are adequate for drug monitoring in clinical research. PMID- 12541821 TI - [Extraction and analysis of profenofos residue in tomato and cabbage by gas chromatography-flame photometric detector]. AB - A quick and effective extraction and clean-up method of profenofos residue in tomato and cabbage is presented. Tomato and cabbage samples were homogenized with a mixture of acetone-hexane (1:1, V/V) using a mechanical homogenizer. The resultant homogenate was cleaned-up by adding active carbon and then filtered under reduced pressure. The filter cake was extracted twice with the same solvent mixture. The filtrates were combined and transferred into a separatory funnel. The organic layer was separated and evaporated to dryness using a rotary evaporator. The residue was dissolved in 2 mL of acetone and transferred into a small glass vial and then determined by GC on a 5% OV-101 Chromosorb W-HP column with flame photometric detector. The results showed that this analytical method can be used for an accurate determination of profenofos residues in tomato and cabbage. The minimum detectable concentration of profenofos in samples was 0.06 mg/kg. The recoveries of profenofos in tomato and cabbage were in the range of 96.2%-105.9% and 94.7%-102.3%, respectively. The relative standard deviations were in the range of 3.7%-4.9% and 3.7%-5.0%, respectively. The tomato and cabbage samples were collected 3 weeks after applying profenofos in the field, and the contents of profenofos were determined. The average contents of profenofos in tomato and cabbage were (13.8 +/- 0.8) mg/kg and (14.4 +/- 0.7) mg/kg, respectively. PMID- 12541822 TI - [Rapid determination of ethephon residues in concentrated pineapple juice by head space gas chromatography]. AB - The method developed is based on the special property that ethephon can be easily decomposed into ethene in alkaline solution by heating. Certain amount of concentrated KOH solution was added to the sample and heated at 70 degrees C. Then, 1 mL of the gas above the sample was injected into a gas chromatograph by head-space sampler for the determination of the target compound with external standard quantitation method. The detection limit was 0.025 mg/kg and the fortified recoveries of ethephon in concentrated pineapple juice (60 +/- 1) Brix at the range of 0.1 mg/kg-10 mg/kg were 92%-98% (n = 8, for each level). The relative standard deviations were 3.99%-7.94%. PMID- 12541823 TI - [Anatomy structures of nasal cavity and paranasal sinus on virtual endoscopy and coronal image]. AB - OBJECTIVE: To evaluate normal and bony anatomy of nasal cavity and paranasal sinus on the spiral CT virtual endoscopy(VE) and coronal scanning image. METHOD: After patients were scanned on axial or coronal position by spiral CT, data were transferred to workstation. Structures such as ostium-meatal complex(OMC) were viewed by navigator software when threshold was changing. RESULT: Turbinates, meatus and ostium-meatal complex were better viewed by virtual endoscopy compared with by coronal scanning image. To some extent, bony anatomy was clearer. CONCLUSION: Virtual endoscopy is of value in realizing characteristics of nasal cavity and paranasal sinus anatomy. PMID- 12541824 TI - [The infiltration and activation stage of eosinophils in nasal polyps]. AB - OBJECTIVE: To assess the infiltration and activation of eosinophils in nasal polyps. METHOD: Nasal polyps from sixteen patients were investigated. The samples were stained with Chromotrope 2R histochemical and ABC immunohistochemical methods. RESULT: The majority of eosinophils in nasal polyps were activated, and there were no significant differences in the numbers of total eosinophils (Chromotrope 2R positive cells) and activated eosinophils (EG2 positive cells) between allergic and nonallergic patients. CONCLUSION: Eosinophilia is a prominent histologic feature of nasal polyps which indicates that the activated eosinophils may play vital role in the pathogenesis of nasal polyps. PMID- 12541825 TI - [The diagnosis and treatment of isolated sphenoid sinus disease]. AB - OBJECTIVE: To detail the underlying symptoms and signs of patients with isolated sphenoid sinus inflammatory disease and enhance the initial diagnostic accuracy. METHOD: 49 patients with isolated sphenoid sinus inflammatory disease were reviewed, and the outcomes of the endoscopic surgery were compared with the non endoscopic surgery. RESULT: In patients who underwent endoscopic sphenoidotomy, 31 were virtually free from symptoms and 5 still experienced mild headache and hyposmia but their radiographic finding had become normalized. None of them need re-operation within 6-48 months follow-up. However, in patients with non endoscopic sinus irrigation, only one was free from the symptoms and 5 patients underwent re-operation because of recurrence. CONCLUSION: Nasal endoscopy and CT/MRI scan could provide a precise diagnosis for isolated sphenoid sinus disease and endoscopic surgery is valuable in its treatment. PMID- 12541826 TI - [The effects of BDP on the inflammatory cells in nasal secretions of the patients with allergic rhinitis]. AB - OBJECTIVE: To study the effects of beclomethasone dipropionate (BDP) on the inflammatory cells in nasal secretions of the patients with allergic rhinitis. METHOD: The variation of intercellular RNA and RNA/DNA ratio of the eosinophils, neutrocyte and lymphocyte in the nasal secretions were observed under the laser scanning confocal microscopy and technology of fluorescence. RESULT: The scanning images displayed that either in eosinophils or in neutrocyte, the fluorescence signal of intercellular RNA was significantly decreased after BDP treatment, and the intercellular RNA/DNA ratio was significantly reduced (eosinophils group P < 0.001, and neutrocyte group P < 0.01). The fluorescence singal of lymphocyte intercellular RNA and RNA/DNA ratio were not significantly changed (P > 0.05). CONCLUSION: BDP can reduce the content of intercellular RNA of the eosinophils and neutrocyte. The anti-inflammation function of BDP is realized by regulating the intercellular RNA synthesize and/or RNA degradation, and by inhibiting the intercellular inflammatory medium production. BDP does not directly influence the intercellular RNA metabolize of the lymphocyte. PMID- 12541827 TI - [Treatment of nuclein 32P colloid perfusion with chronic suppurative maxillary sinusitis]. AB - OBJECTIVE: To determine the effect of nuclein 32P colloid in treatment of chronic maxillary sinusitis (CMS). METHOD: 22 patients (31 sides) with CMS after washing of sinucavity were perfused with nuclein 32P in sinucavity. 20 patients (31 sides) with routine conservative treatment were served as a control group. Two groups were followed up for one year. RESULT: In nuclein group, 27 sides were cured, 4 sides were recurred and the rate of cure was 87%. In routine group, 19 sides were cured, 12 sides were recurred and the rate of cure is 61%. There is significant difference between the two groups (P < 0.01). The treatment active of nuclein group is better than that of routine group. CONCLUSION: Nuclein 32P colloid with the function of ionizing radiation can increase the pathogenic tissue immunocompetence and metabolic course, obstruct the microvascular and promote the absorption of inflammatory exudation and edema subsidence. It can also make the proliferous tissue atrophy thence accelerate the rate of cure of CMS. PMID- 12541828 TI - [The preliminary research of maxillary sinus cancer on oncogene level]. AB - OBJECTIVE: To determine the oncogene c-myc, bcl-2' product expression in the maxillary sinus cancer and to investigate its clinical significance. METHOD: In 46 maxillary sinus cancer and 56 normal nasal mucous membrane, immunochemical method was adopted to determine oncogene c-myc and bcl-2' product. RESULT: In the normal nasal mucous membrane, the positive rates of oncogene c-myc and bcl-2' product were 0; in the maxillary sinus cancer, the positive rates of c-myc and bcl-2' product were 73.91 (34/46), 60.87 (28/46) respectively. The positive rate of c-myc' product in the relapse group(91.67%, 11/12) was significantly higher than that in the non-relapse group(67.65%, 23/34), P < 0.01; the positive rate of bcl-2' product in the relapse group(50%, 6/12) was significantly lower than that in the non-relapse group(64.71%, 22/34), P < 0.05; the positive rate of c-myc' product in the stage IV (88.89%, 8/9) was significantly higher than that in the stage II (63.64%, 7/11), P < 0.05; the positive rate of bcl-2' product in the stage II (72.73%, 8/11) was significantly higher than that in the stage IV (55.56%, 5/9), P < 0.05; the positive rate of bcl-2' product was in the negative correlation to the pathological grade of primary carcinoma. CONCLUSION: The oncogene c-myc and bcl-2 take part in the genesis and advancing course of maxillary sinus cancer. Their products were the important oncogology indexes, which might be useful to determine illness condition and predict relapse. PMID- 12541829 TI - [Detection of bate 2-microglobulin fibronection and interleukin 2 content in patients with nasopharyngeal carcinoma]. AB - OBJECTIVE: To study the clinical significance of detecting of beta 2 microglobulin (beta 2-MG). Fibronection (Fn) and interleukin 2 (IL-2) in patients with nasopharyngeal carcinoma. METHOD: The levels of serum beta 2-MG and IL-2 in 32 patients with nasopharyngeal carcinoma were determined by radioimmunoassay, radial immunodiffusion and ELISA before and after radiotherapy. RESULT: The levels of serum beta 2-MG in patients with nasopharyngeal carcinoma were significantly higher than that in controls (P < 0.05); but serum Fn and IL-2 in patients were significantly lower than that in controls (P < 0.05). The level of serum beta 2-MG decreased in patients after radiotherapy, but the serum Fn and IL 2 increased in patients after radiotherapy. CONCLUSION: Detection of serum beta 2 MG, Fn and IL-2 was useful in clinical diagnosis and prognosis observation of nasopharyngeal carcinoma. PMID- 12541830 TI - [Detection of radiation-induced apoptosis in nasopharyngeal carcinoma]. AB - OBJECTIVE: To investigate the pathological changes after radiotherapy and its clinical significance of nasopharyngeal carcinoma (NPC). METHOD: Paraffin embedded tissue blocks obtained from 30 cases of NPC before and during radiotherapy were used in this study. Using terminal oxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL) and HE staining, the spontaneous and radiation-induced apoptotic rate (AR) were examined, and the pathological changes of NPC tissues before and after radiotherapy were compared. RESULT: The AR of radiation-induced apoptosis was significant higher than that of spontaneous apoptosis, but the AR was decreasing and the differentiation of proliferatic tumor clonogehic cell getting better during radiotherapy of NPC. CONCLUSION: The process of apoptosis was accelerated and the differentiation of proliferative tumor clonogehic cell getting better during radiotherapy of NPC. PMID- 12541831 TI - [Study on some metal elements levels and the T lymphocyte subset of the peripheral lymphocytes in patients with nasopharyngeal carcinomas]. AB - OBJECTIVE: To study some metal elements levels and T lymphocyte subset of the peripheral lymphocytes in patients with nasopharyngeal. METHOD: Metal elements in the patients with nasopharyngeal and normal control were detected by the Atomic Absorption Spectrometer (Flame SHIMATZV.JAPAN). T lymphocyte subset were detected by the flow cytometry (FCM). RESULT: The level of copper was significantly higher and the level of zinc was significantly lower in patients with nasopharyngeal carcinomas than those in normal control group (P < 0.01 and < 0.05 respectively). The number of CD3/CD4 cell in the patients group were less than those in normal group(P < 0.01). CONCLUSION: The results of correlative analysis showed that there was a significant correlationship between the number of CD4 cells and levels of copper in peripheral lymphocytes of the patients group. PMID- 12541832 TI - [Changes of NK cell's activity and T lymphocyte subpopulation in peripheral blood of patients with laryngocarcinoma]. AB - OBJECTIVE: By detecting the changes of NK cell's activity and T lymphocyte subpopulation in the peripheral blood of the patients with laryngocarcinoma, the immune function level of the patients could be judged and the prognosis of the patients could be predicted. METHOD: The NK cell's activity in 47 patients with laryngocarcinoma was tested, and T lymphocyte subpopulation in 35 patients were detected with flow cytometer, and compared with the normal control group of 45 volunteers and the group of 15 patients with polyp of vocal cord. RESULT: The NK cell's activity in the patients with laryngocarcinoma was lower than both of the normal control group and the polyp of vocal cord patients group, and the more severe was the degree of disease, the lower of the NK cell's activity was. The NK cell's activity in the pharyngocacinoma patients group in the 3-6 days after operation was lower than that in preoperation significantly; in the same time when the NK cell's activity was droping, CD3 and the ratio of CD4/CD8 became lower. CONCLUSION: By detecting the changes of both NK cell's activity and the T lymphocyte subpopulation in patients with laryngocarcinoma, the immune function level of the patients can be judged objectively, which is of the important clinic significance. PMID- 12541833 TI - [An analysis of the results of central masking effect on healthy ears]. AB - OBJECTIVE: To investigate the mechanism of Central Masking Effects(CME) and its effects on audiometry measurement. METHOD: The pure tone threshold was measured when the notest ear was masked or was in quiet respectively. We investigated the different results while giving different level maskers at the same frequency, or giving the same masker at different frequencies to find out their rules. RESULT: The CME has frequency-selective properties and sound level-selective properties. The CME appears apparently at 1 kHz and 2 kHz. At 2 kHz, the CME get the highest. When the masker was lower than 60 dB HL, the CME became higher as the masker was tuned higher; when the masker was 60 dB HL, the CME get the highest[(11.53 +/- 4.38)dB HL]. If the masker was higher than 70 dB HL, overmasking appeared. CONCLUSION: Our observations come to the conclusion that central masking correction should be made clinically when the masker is higher than 40 dB HL. 60 dB HL masker at 2 kHz can make the CME higher than 10 dB HL, thereby can be used to identify false deafness. PMID- 12541834 TI - [Transmission electron microscopy of the tegmentum vasculosum in chick inner ear]. AB - OBJECTIVE: To study the ultrastructure of the tegmentum vasculosum(TV) in chick inner ear so as to obtain important information for its physiology and pathology. METHOD: The ultrastructures of TV in 20 chicks were observed by transmission electron microscope(TEM). RESULT: TV was composed of flask-shaped dark cells, which had dense cytoplasm; hourglass-shaped light cells, which had light cytoplasm; abundant microvascula. Dark cell had the extensive networks of folded plasma membrane along the side and throughout the lower half of the cell, these folds of dark cell interdigitated with thin processes which extended from the lateral walls of light cell, these were sparse microvilli on the luminal surface of light cell and sparse cellular organizations in light cell. Light cell had more direct contacts with the micrangium. CONCLUSION: TV is very similar to stria vascularis but there are also some differences between them under TEM. PMID- 12541835 TI - [Study on quantitative relationship between retention parameters of solutes in reversed-phase liquid chromatography]. AB - A quantitative relationship between lg I (a constant relating to the affinity of one mole of solute to stationary phase) and Z (total moles of organic solvent released at the contact region as one mole solute adsorbed by stationary phase) in the stoichiometric displacement model for retention (SDM-R) of solute was compared with the corresponding relationship in Snyder's Equation in reversed phase liquid chromatography (RPLC). It was found that the relationship from SDM-R was more exact than that from Snyder's Equation. The linearity depends on the ranges of both lg I and lg Kw0. The broader the range, the better the linearity is. This conclusion conflicts to that reported in references in which the linearity only exists either when one factor dominates solute retention, or when it is completely dominated by statistics. The linear relationship in SDM-R and in Snyder's Equation was found valid not only for homologues but also for non homologues. PMID- 12541836 TI - [Syntheses of polymer coated silica packing materials and their retention behaviors for reversed-phase liquid chromatography]. AB - The polymer coated silica packings C or D with methylmethacrylate(MMA) or MMA ethylene dimethacrylate(EDMA) monomer were prepared. For the packing materials the polymer coated degree, effects of cross-linking agent and retention behavior were characterized by diffuse reflectance infrared Fourier transform spectroscopy, elemental analysis, size exclusion chromatography and reversed phase liquid chromatography. The results showed that polymer coated degree on silica depended on the amount of monomer, and the volume of micropore of the meterials, which was controlled more by the cross-linked structure than by the linear structure. The hydrophobicity, retention ratio and number of theoretical plate (80% methanol and 60% acetonitrile) of the packing materials in comparison with those of C18 are discussed. The polymer coated silica packings C and D showed the same stereoselectivity for separation of aromatic compounds, such as triptycene. PMID- 12541837 TI - [The effect of temperature on the enantiomeric resolutions on albumin and beta cyclodextrin chiral stationary phases]. AB - The effect of column temperature on enantiomeric resolutions of tryptophan, warfarin and ketoprofen was investigated on bovine and human serum albumin (BSA and HSA) stationary phases, which were synthesized with s-triazine as the activator. It was observed that the entropy change made a great contribution to the separation of those enantiomers on albumin chiral stationary phases. The column temperature for the maximal resolution of tryptophan on the BSA stationary phase prepared by this method was about 35 degrees C, which was not 24 degrees C as reported for the BSA stationary phase synthesized with glutaric dialdehyde as the activator. This results may come from the different conformation of the immobilized BSA and HSA due to the different coupling methods used. On the other hand, it was indicated that the resolution of enantiomers on beta-CD chiral stationary phase was mainly contributed from the change of enthalpy, which means the resolution of chiral solutes on albumin and beta-CD stationary phases has different thermodynamic behaviors. PMID- 12541838 TI - [Determination of enantiomeric purity for lactic acid in fermentation broth by Rhizopus oryzae with high performance liquid chromatography]. AB - A procedure for the resolution of DL-lactic acid and the determination of D isomer ratio in L-lactic acid fermentation broth by Rhizopus oryzae is described. The effects of pH of mobile phase and concentration of chiral mobile phase additives on resolution of DL-lactic acid were investigated. The optical isomers of lactic acid were resolved by RP-HPLC with 2,3,6-tri-O-beta-cyclodextrin(TM beta-CD) as a chiral mobile phase additive, and C18 column as stationary phase, and detected at wavelength 210 nm. The results showed that a correction factor should be introduced into the equation for calculation of the percentage of D lactic acid, because the UV absorption of D-lactic acid and L-lactic acid might not be the same when TM-beta-CD was present. Quantitation was achieved with external standard method, the average recovery was 100.4%, and the relative standard deviation was 0.82%. This method can be used for the determination of the percentage of D-isomer in L-lactic acid fermentation broth by Rhizopus oryzae, and it is simple, rapid and accurate. The results showed that the mass fraction of D-isomer in the fermented broth increased during the period of storage. PMID- 12541839 TI - [Study of the adsorption properties in chromatographic separation of the citric acid]. AB - A new method studying the change of thermodynamic properties at different temperatures to screen suitable stationary phases for chromatographic separation of the citric acid (CA) on an analytical column has been developed. The applied method is mainly based on using CA with a series of solute concentrations (0 g/L 250 g/L) as mobile phase to seriously overpass the unloaded stationary phase to find the breakthrough curves of CA. The adsorption isotherms were calculated from the retention times of breakthrough curve. The bed porosity of column was calculated from the elution time of a peak of the blue dextran as non-adsorbed tracer. The stationary phase, which has been selected through this method, for the separation of CA from fermentation broth, showed a very high adsorption capacity for CA, even under very low concentrations, so it can even be used in the separation and purification process of minute amounts of CA. The separation effect of this stationary phase for citric acid can be greatly enhanced through the technique of change of temperature. This new separation process can shorten the old process path, improve the yield of finished products more than 10%, and reduce the cost more than 10%. At the same time, the pollution of the production of citric acid can be reduced. It turned out that this method will play an important role in the synthesis of stationary phase for the separation of citric acid and the study of its structure. PMID- 12541840 TI - [Preparation of novel magnetic dextran affinity adsorbents and their application to purify urokinase]. AB - The reverse phase suspension and embedment technique were adopted to prepare magnetic dextran microsphere (MDMS). The dispersion medium was mixture of some organic solvents. Span-80 was used as stabilizer. The aqueous dextran with magnetic fluid was suspended in dispersion medium with epichlorohydrin as cross linking reagent. The mixture was stirred for 30 minutes at room temperature and then heated at 70 degrees C for 4 hours, MDMS was thus obtained. MDMS was activated by epichlorohydrin on which 6-aminohexanoic acid, glycine or ethylene diamine was bonded as spacers. Then it was coupled with p-aminobenzamide, L arginine methyl ester or guanidohexanoic acid and five magnetic affinity adsorbents were prepared. The MDMS was polydisperse particles with the size of 50 300 meshes and the content of Fe3O4 was about 6.2 per cent in the MDMS. Influence of some parameters such as viscosity and density of organic phase, the volume ratio of organic and aqueous phase, the quantity of surfactant and stirring speed on preparing MDMS was studied. Magnetic affinity adsorbents were used to purify crude urokinase in a bath mode and the effect of coupling reagents and ligands on results of purification was discussed. The bioactivity recovery was 40.0 to 60.7 per cent, the purification-fold was between 14.9 and 32.8, and the adsorptive capacity varies from 89 mg to 121 mg per milliliter of adsorbent. PMID- 12541841 TI - [Estimation of gas chromatographic retention index for polycyclic aromatic hydrocarbons using VMDE]. AB - A novel molecular distance-edge vector with 6 parameters was calculated for 100 polycyclic aromatic hydrocarbons (PAHs) based on the molecular interaction theory. It was found that there was a good linear relationship between the molecular distance-edge vector (VMDE) and the gas chromatographic retention index for the 100 PAHs, and the molecular modelling equation for the quantitative structure-retention relationship (QSRR) can be expressed as follows: IR = a + b VMDE, n = 100, r = 0.988, where a and b represent regression coefficients. The regression results showed that the molecular distance-edge vector is in good agreement with the results of molecular polarizability index to the gas chromatographic retention index for the polycyclic aromatic hydrocarbons. PMID- 12541842 TI - [The design and evaluation of horizontal pipe mini-flame atomization and ionization synchronous detector in GC/AAS]. AB - Gas chromatography/atomic absorption spectroscopy(GC/AAS) is a good method for the species analysis of organometallic compounds. But the traditional atomizers are not very suitable for this technology and all of them response only to one kind of signal--the concentration of the metallic atoms of the ground state. They can not give any information about the organic group of organometallic compounds and organic compounds which coexist with the former. For GC/AAS we want to design and manufacture a new kind of detector which is much more sensitive and has a much smaller dead volume and will sensitively and synchronously response to the atomization signal of organometallic compound and ionization signal of organic compound. The authors have noticed that the atomization of organometallic compound and ionization of organic compound have been existing in the same hydrogen flame. The question is how to gain and exchange and output the two signals which are completely different in characters. For this purpose we designed and manufactured a new type of horizontal pipe mini-flame atomization and ionization synchronous detector. The key part is a T type glass tube (80 mm x 13 mm x 10 mm i.d.) which covers horizontally on the jet of the mini-flame atomizer and a long pipe stainless steel collector (70 mm x 9.5 mm o.d. x 9 mm i.d.) is tightly inserted in the tube. The light beam of the hollow cathode lamp passes through the hydrogen flame along the axial center of the glass tube and the ground state metallic atoms in the flame diffuses to both ends of the tube along the axial center of the glass tube too. This process enriches the depth of the light absorption and then increases the sensitivity of atomization. At the same time the long pipe collector can enrich the collection efficiency of the ionization and increases the sensitivity and widens the linear range. So the detector can synchronously and sensitively detect the organometallic and organic compounds. The detection limit for atomization of the organometallic compound(diethylmercury) is 2.5 x 10(-11) g.s-1 and for ionization of the organic compound (benzene) is 1.0 x 10(-11) g.s-1. The linear range is 7.0 x 10(2) for atomization of diethylmercury (r = 0.9989) and 2.5 x 10(5) for ionization of benzene (r = 0.9992). The selectivity of the detector to the organic solvent under the optimum operating condition of atomization is 10(4)-10(7). The relative standard deviation (RSD, n = 11) of the peak area in the optimum operating conditions of atomization for the organometallic compound (diethylmercury) and of ionization for the organic compound (benzene) is 1.8% and 1.5% respectively. Under the same operating condition, the atomization and the ionization can not reach the optimum performance at the same time. If the atomization is in the optimum performance state, the performance for ionization is like a chromatographic thermal conductivity detector (TCD). The optimum performance of the atomization and ionization can be quickly adjusted by changing the flow rates of the corresponding gases such as H2, N2 and air. PMID- 12541843 TI - [Advances in chromatography and related techniques in China--status report in 1998-1999]. AB - This report covers developments in chromatography and related techniques in China during the period of 1998-1999. The papers published in 23 domestic and a few foreign periodicals which are closely related to chromatography, and presented at 1999 Beijing Conference and Exhibition on Instrumental Analysis and the 12th National Symposium of Chromatography are reviewed. Specific topic areas covered include gas chromatography (GC), high performance liquid chromatography (HPLC), capillary electrophoresis (CE) and capillary electrochromatography (CEC), solid phase extraction (SPE) and solid phase microextraction (SPME). PMID- 12541844 TI - [The synthesis of phenyl-silica gel and its application in purification of taxol]. AB - Phenyl-silica gels were synthesized with four methods of reaction on gamma aminopropyltriethoxysilane (APTS) silica gel. The synthesized gels were used as stationary phase of normal pressure chromatography to separate and purify taxol. The comparison of these four silica gels was carried out. It was found that the gel synthesized by gas-phase reaction gave the best results. The purity of taxol was 77.4%, and the recovery was about 90%. PMID- 12541845 TI - [Optimized linear temperature programming for capillary gas chromatography by the method of gridding search]. AB - Linear temperature programming of capillary gas chromatography was optimized by using a new method of gridding search. A point of gridding demonstrated a condition of temperature programmed and the resolution and the retention time were optimized at all gridding points. With a new design of calculation, the method not only considered directly all components that were separated, but the velocity of calculation was improved also. Twenty-one compounds of alkanes, alcohols, halo-hydrocarbons and aromatic hydrocarbons, of which Kovats Indexes are varying from 600 to 1,000, were separated by this new method on an OV-101 column under linear temperature programming. Good results were obtained. The time for optimizing the 21 compounds was less than 1 min. PMID- 12541846 TI - [Determination of surface energy of PS-PEO-PS triblock copolymers by inverse gas chromatography]. AB - The surface energy(gamma sd) of the dispersive component of the triblock copolymers, poly(styrene)-poly (ethylene oxide)-poly(styrene)(PS-PEO-PS), was determined by using inverse gas chromatography(IGC). A series of linear alkanes were used to determine the free energy of adsorption (delta GCH2) of a CH2 group in a temperature range of 70 degrees C-120 degrees C, and gamma sd of PS-PEO-PS were calculated. The effects of the polymer composition and temperature on gamma sd values were discussed. IGC data indicated that gamma sd values of PS-PEO-PS were lower (gamma dPS-PEO-PS-1 = 16.99 mJ.m-2, gamma dPS-PEO-PS-2 = 17.98 mJ.m-2, 70 degrees C) than that of PEG (gamma dPEG = 21.74 mJ.m-2, 70 degrees C), and increased as the ethylene oxide content increased in PS-PEO-PS. The gamma sd was sensitive to temperature, decreased sharply with the increase of temperature. Linear relationships were obtained between the gamma sd and temperature (t, degree C), which were gamma dPE-PEO-PS-1 = 28.12-0.161t and gamma dPS-PEO-PS-2 = 28.23-0.148t. PMID- 12541847 TI - [Determination of triptolide in human plasma by gas chromatography-electron capture detection]. AB - A method for the determination of tiptolide in human plasma by gas chromatography electron capture detection is described. An SE-54 capillary column (30 m x 0.25 mm i.d.) was used. Nitrogen was used as the carrier gas. After NaOH solution being added, the plasma was extracted with ether-chloroform (3:1, V/V) and then reacted with trifluoroacetic anhydride. A good linearity was obtained from 1.0 microgram/L to 50.0 micrograms/L of tiptolide in human plasma, r = 0.9990. The detection limit of it in plasma was 0.5 microgram/L. The average recovery was 96.3%. The method is sensitive and precise. PMID- 12541848 TI - [Electrophoretic migration behavior of deoxyribonucleic acid fragments in three polymer solution concentration regions]. AB - The theory of polymer coils shrinking in semi-dilute solution has recently been developed on polymer solution. The polymer solution from coils shrinking concentration Cs to uniform entangled concentt concentration C+ has been defined as semi-dilute solution. We experimentally investigated the electrophoretic migration behavior of 100 bp deoxyribonucleic acid (DNA) Ladder in hydroxypropyl methyl cellulose (HPMC) concentration ranging from 1.25 g/L to 16.06 g/L. The friction force mobility mu f is used to express the friction force that DNA will encounter in capillary electrophoresis. Our results indicate the division of polymer solution into three regions depending on the relationship of mu f and HPMC concentration and Ferguson plot. Resolutions of 200 bp/300 bp and 700 bp/800 bp show semi-dilute polymer solution suits large fragments and small fragments DNA separation respectively. the results confirm that the current polymer theory is valid under actual CE condition. PMID- 12541849 TI - [Determination of the pKa value of icariin and its content in the Chinese herb medicine epimedium grandiflorum morr. by capillary electrophoresis]. AB - Knowledge of dissociation constants is important for prediction and understanding the migration behavior of analytes in capillary electrophoresis. Icariin is the active component of the Chinese herb medicine Epimedium grandiflorum Morr.. In order to determine the dissociation constant of icariin and to show many important pharmacology activities, a CZE method has been used to determine the ion mobility (mu A-) and the pKa value of icariin based on the non-linear relation between mu eff and [H+] and the linear relation between the reciprocal of effective mobility of the solute (1/mu eff) and the [H+] of the buffer solution. In addition, the change of pKa of icariin with the increase of ethanol concentration in the buffer was also investigated. Under the buffer condition of 24 mmol/L phosphate + 30% ethanol, the content of the active component icariin in Chinese herb Epimedium grandiflorum Morr. was quantitatively determined. The linear correlation equation was Y = 6.96 x 10(-3) + 17.0X, and the linear range is 0.032 g/L-0.354 g/L. PMID- 12541850 TI - [Determination of angiotensin-converting enzyme activity by micellar electrokinetic capillary chromatography]. AB - A sensitive and rapid method was developed for angiotensin-converting enzyme (ACE) activity determination by micellar electrokinetic capillary chromatography (MECC). MECC was carried out to separate and quantify the products of the enzymatic reacting using Hip-Leu-His as the substrate in 20 mmol/L boric acid borate buffer (pH 9.0) including 50 mmol/L SDS as the run buffer at an applied voltage of 8.1 kV. The electrophoresis was monitored at 228 nm, and completed in 6 minutes. The detection limits of ACE activity was 5 pmol/min(signal to noise ratio was 2). PMID- 12541851 TI - [Dynamic pre-concentration method of gas sample]. AB - By utilizing the combination of frontal chromatography at ambient temperature, back-flushing and temperature gradient thermal desorption, a prototype of a dynamic pre-concentration system has been designed and evaluated. It realized the enrichment for gas samples of boiling point higher than - 103 degrees C without cryogenic facilities. To reduce the sample volume and enhance enrichment factor, the technique of momentarily non-steady state produced at the beginning of back flushing, and the negative temperature gradient generated during thermal desorption were utilized, resulting a very sharp desorption band. Standard samples were tested to demonstrate the applicability of the method. At 10 mL sample volume, the enrichment factor was 100. PMID- 12541852 TI - [Novel affinity membrane used for bilirubin removal]. AB - Bilirubin is a toxic substance. In order to effectively remove it from the hepatic patients' blood, two novel affinity membranes were prepared. These were prepared by chemically grafting on cellulose and immobilized with different ligands. One kind of ligand was poly-D-lysine, the other one was quaternary ammonium salt. Both affinity membranes were used for removal bilirubin from phosphate buffer and HSA solutions, and the effects of temperature, HSA concentration, adsorption time in static state experiment and flow rate in dynamic state experiment have been investigated. The results indicated that the membranes could remove over 70% bilirubin from phosphate buffer and at least 50% from low concentration HSA solutions. The results also indicated that the removal efficiency was better at higher temperature. In the static state experiment, four hours can be selected as adsorption time. In the dynamic state experiment, the flow rate can be properly increased. PMID- 12541853 TI - [Analysis of main chemical composition in hydrogenated rosin from Zhuzhou]. AB - The acid fraction, the main part of the hydrogenated rosin produced by Zhuzhou Forest Chemicals Plant of China, was separated from neutral fraction by modified DEAE-Sephadex ion exchange chromatography and analyzed with GC-MS-DS technique by using DB-5 capillary column. Six dihydroabietic-type resin acids, four dihydropimaric/isopimaric-type resin acids and four tetrahydroabietic-type resin acids were identified. The hydrogenated rosin is composed mainly of 8-abietenoic acid, 18-abietanoic acid, 13-abietenoic acid, 8 alpha, 13 beta-abietanoic acid, 13 beta-8-abietenoic acid and 8-isopimarenoic acid etc. PMID- 12541854 TI - [Analysis of chlorogenic acid in sunflower cake by thin-layer chromatography]. AB - The analytical method of chlorogenic acid in sunflower cake by thin-layer chromatography(TLC) was developed. With 36% acetic acid as mobile phase and on the polyamide film plate, the Rf value of chlorogenic acid was 0.61. The calibration curve was linear in the range of 0.05 microgram-0.6 microgram of chlorogenic acid with the detection limit of 0.025 microgram and the average recovery of 97.53%. The inter-plate accuracy of chlorogenic acid was 2.57%. With 70% ethyl alcohol as extracting solvent and 30 min of extraction time, the amount of chlorogenic acid in the extract, the residue amount of chlorogenic acid in the sunflower cake and the extraction rate were determined and calculated by applying this method. The optimal time of extraction was found. PMID- 12541855 TI - [Determination and purification of beta-carotene in Spirulina maximum]. AB - There are several kinds of nutritious materials in Spirulina maximum, and beta carotene is one of the most important materials. Because beta-carotene is of low polarity, for purification it was extracted with the mixture of acetone and methanol in the ratio of 7:2 (V/V), then saponified, and extracted with petroleum ether, purified with column packed with neutral Al2O3. Its content was determined by reversed-phase HPLC. This method is accurate and the results are reproducible. The average recovery was 96.8%. PMID- 12541856 TI - [Determination of degraded products of penicillin by high performance liquid chromatography]. AB - A method for the determination of main composition in degraded products of penicillin, 6-aminopenicillanic acid (6-APA), penicillin G kalium (PGK), phenylacetic acid (PAA), and the by-product benzylpenicilloic acid (BPA) by HPLC was studied. Separation conditions were as follows: Spherisorb C18 column, 250 mm x 4.6 mm i.d., 10 micrograms; mobile phase: V (methanol): V(0.004 mol/L KH2PO4 buffer, pH 4.5) = 50:50; flow rate: 1 mL/min; detector: UV 230 nm. The method is effective, quick, accurate and reproducible. The satisfactory results show that this new method has certain practical values as an approach of quality control. PMID- 12541857 TI - [Detection of residual antibiotics in honey with capillary electrophoresis]. AB - Five antibiotics compounds, tetracycline (TC), oxytetracycline (OTC), doxycycline (DOC), chlortetracycline (CTC) and chloramphenicol (CP), were successfully separated and determined by high performance capillary electrophoresis(HPCE). Effects of buffer pH, various organic additives and temperature on electrophoretic separation of antibiotics were investigated. Satisfactory separation of these five antibiotics was achieved in the buffer of pH 3.2, 0.02 mol/L Na2HPO4-0.01 mol/L citric acid with addition of 4% (V/V) N-methylmorpholine and 12% (V/V) acetonitrile within 20 minutes. The calibration graphs were linear by plotting the peak area against the sample concentration over the range of 150 micrograms/L to 750 micrograms/L and the correlation coefficients were greater than 0.9917. The detection limits were 10 micrograms/L for CP, 20 micrograms/L for TC, OTC and DOC and 40 micrograms/L for CP (signal to noise ratio > 5). HPCE method was successfully applied to the analysis of trace antibiotics in honey. PMID- 12541858 TI - [Determination of micro amounts of iodide in dairy products by ion chromatography with direct amperometric detector]. AB - A method was developed for the determination of micro amounts of iodide in dairy products by ion chromatography with direct amperometric detection. The eluent used was 0.04 mol/L NaOH. The relative standard deviations of retention times and peak heights for 1 mg/L and 2 micrograms/L iodide solutions were 0.67%, 3.9% (n = 8) and 0.97%, 1.8% (n = 8), respectively. Calibration curves showed good linearity with r = 0.9991 (n = 8, from 10 micrograms/L to 160 micrograms/L), and with r = 0.9986 (n = 8, from 400 micrograms/L to 5 mg/L). The detection limit of iodide by this method was 1 microgram/L. The method has been used to analyse two kinds of milk powder and a kind of fresh milk. The recoveries were 89.0%, 86.0% and 84.1% for infant formula milk powder, mother's milk powder and fresh milk, respectively. PMID- 12541859 TI - [Expression of Th1, Th2-typed cytokines and its significance in nasal polyps]. AB - OBJECTIVE: To evaluate the possible role of cytokines in pathophysiology and treatment of nasal polyps. METHOD: The expressions of Th1-typed cytokines IFN gamma, IL-2, IL-12 and Th2-typed cytokines IL-4, IL-5, IL-8, IL-10, IL-13 were investigated with enzyme-linked immune sorbent assay(ELISA) in 25 patients with nasal polyps. RESULT: There was a significant upregulation of Th2-typed cytokines IL-4, IL-5, IL-8, IL-10, IL-13 in nasal polyps compared with normal nasal mucosa, especially IL-4 and IL-5 (3.9 times and 8.8 times higher than normal mucous respectively) while the expression of Th1-typed cytokines IFN-gamma, IL-2, IL-12 decreased after treated with local glucocorticoid. The levels of Th2-typed cytokines decreased significantly and Th1-typed cytokines had no obvious change. CONCLUSION: The upregulation of Th2-typed cytokines such as IL-4, IL-5, IL-8, IL 10, IL-12 may play an important role in the pathophysiology of nasal polyps and Th2-typed cytokines can be viewed as a target of treatment to nasal polyps. PMID- 12541860 TI - [Observations of the curative effect with various liquid for post operative irrigation of ESS of treating chronic sinusitis and nasal polyps]. AB - OBJECTIVE: To explore the influence of liquid for post-operative irrigation on the effect of ESS. METHOD: 192 cases with chronic sinusitis and/or nasal polyps of groupIIstageII who had undergone the ESS from September, 1998 to June, 1999 and completed a 3-month follow-up were divided randomly into 3 groups to accept 3 different kinds of irrigation liquid. The evaluation of curative effect was according to the index of HaiKou ESS-97. RESULT: The curative effect of Herba Houttuyniae group was better than the other two. CONCLUSION: The post-operative care is important and the irrigation liquid can affect the effect of ESS. PMID- 12541861 TI - [The analysis of long-term curative effect in 201 cases of endoscopic sinus surgery]. AB - OBJECTIVE: To analyze the long-term curative effect of endoscopic sinus surgery (ESS), and to study the factors related to the defeat of endoscopic sinus surgery. METHOD: 201 cases (353 sides) of chronic sinusitis and nasal polyps were treated with ESS, and were treated by taking the nasal cavity as a whole unit. RESULT: Following up survey from 12 months to 24 months postoperatively, it was found that 108 cases were cured (53.7%), 82 cases achieved symptomatic relief(40.8%), 11 cases remained failure (5.5%). CONCLUSION: It is very important to improve the ESS technique in order to reduce the recurrence of nasal polyps and sinusitis and to improve the curative effect of ESS, in addition, the treatment after operation is also indispensable to successful surgery. PMID- 12541862 TI - [Observation on the changes in nasopharyngeal carcinoma patients' olfactory before and after radiotherapy]. AB - OBJECTIVE: Detecting nasopharyngeal carcinoma (NPC) patients' olfactory before and after radiotherapy for investigating the infection of radiotherapy on these patients' olfactory. METHOD: 100 NPC patients' olfactory were detected before the start of radiotherapy and after the termination of therapy in 3, 6, 12, 24 and 36 months. RESULT: Patients' smell acuity decreased sharply 3 months after therapy, but varying degrees of recovery were noted in 6 and 12 months. However their smell acuity became worse in the following time. CONCLUSION: NPC patients' smell acuity can be profoundly affected by radiotherapy. And after radiotherapy, their smell acuity will decrease wave-likely and cannot fully recover. PMID- 12541863 TI - [The influence of IL-2 on the immunologic function of the NPC patients treated with radiotherapy and chemotherapy]. AB - OBJECTIVE: To investigate the combining therapy which not only have cured effect but also can uphold and improve the NPC patient's immunity function after radiotherapy and chemotherapy. METHOD: 90 cases randomly divided into 3 groups 1. Local group (local injected with IL-2 + radiotherapy + chemotherapy); 2. General group (ivdrip with IL-2 + radiotherapy + chemotherapy); 3. Convention group (radiotherapy + chemotherapy). Checked and observed the immunity function around the immunotherapy and after the radiotherapy and chemotherapy. RESULT: Cellular immunity of 3 groups are lower and humoral immunity are hypetuntion than normal person. After treated with IL-2 the cellular immunity improves but there's no great change of the humoral immunity. The immune status of the immune groups have not obvious change than before radiotherapy, at the same time, the cellular immunity of the convention group cut down and the humoral immunity doesn't change obviously. CONCLUSION: 1. It has some effect to uphold and improve the NPC patient's immunity function to treat with small dosage of IL-2 before radiotherapy and chemotherapy, general treatment is better than local injection; 2. The three therapies have not great influence on the patient's humoral immunity. PMID- 12541864 TI - [DNA character of the cholesteatoma]. AB - OBJECTIVE: To compare the DNA character with in cholesteatoma tissues with that in normal skin. METHOD: DNA content within 25 cholesteatoma tissues and 10 normal postauricular skin was measured with flow cytometry. RESULT: The content of cholesteatoma tissues and postauricular skin specimens are a normal euploid. The cell number during S phase of cholesteatoma (10.9 +/- 2.32)% was higher than that in normal skin(7.31 +/- 2.82)%. CONCLUSION: Although the cholesteatoma has a very high proliferating character, the result suggest it isn' t a malignant tumor. PMID- 12541865 TI - [A clinical analysis of cholesterol granuloma following chronic suppurative otitis media]. AB - OBJECTIVE: To investigate the pathophysiological mechanism, clinical menifestation and radiographic diagnosis of cholesterol granuloma following chronic suppurative otitis media. METHOD: Six cases of CG following chronic suppurative otitis media, confirmed by surgery and pathology, were reviewed and analyzed. RESULT: CG frequently accompanied with other middle ear diseases, and was shown as a high signal intensity on both T1-and T2-weighted images in magnetic resonance imaging(MRI). CONCLUSION: It was postulated that the obstruction of pneumatized temporal bone air cell, caused by other middle ear diseases such as cholesteatoma and tympanosclerosis, might be the pathophysiological mechanism of CG. The evaluations of computed tomography (CT) and clinical manifestation were limited to distinguish CG from cholesteatoma or other neoplasm, while the MRI can be of great value to characteristic diagnosis. PMID- 12541866 TI - [A study on expression of tumor necrosis factor alpha in middle ear cholesteatoma and its effect on bone destruction]. AB - OBJECTIVE: To determine the expression and localization of tumor necrosis factor alpha (TNF-alpha) in human middle ear cholesteatoma and its effects on bone resorption. METHOD: The distribution of TNF-alpha in 18 cholesteatoma tissues and 8 normal external ear canal skin, facial skin and tympanic membrane were studied immunohistochemically with paraffin-embedded sections. RESULT: TNF-alpha was localized in the epithelium and connective tissue of cholesteatomas studied, particularly on basal and spinous cells and macrophages. The epithelium of normal external ear canal skin, facial skin and tympanic membrane was slightly stained. CONCLUSION: This study revealed significantly increased level of TNF-alpha in cholesteatoma tissues. TNF-alpha acts both directly, by causing bone erosion as an autocrine growth factor, and indirectly as an important mediator by stimulating the release of enzymes causing bone destruction. PMID- 12541867 TI - [Effect of arytenoid adduction operation for vocal function of laryngeal palsy patients]. AB - OBJECTIVE: To evaluate the curative efficiency on vocal function of 56 laryngeal palsy patients underwent arytenoid adduction operation. METHOD: Voice analysis and laryngeal aerodynamical measurement were performed for the 56 laryngeal palsy patients to compare maximum phonation time (MPT), mean flow rate (MFR), voice intensity(SPL), and expiratory pressure(EP) before and after operation. Perceptual evaluation was also performed for comparison by using GRBAS system(G). RESULT: The postoperative average MPT increased by 3.1 times in males and 2.7 times in females. The average MFR decreased apparently from 827.3 ml/s in males and 477.1 ml/s in females before operation to 340.3 ml/s and 158 ml/s after operation, which was statistically significant (P < 0.01). The average SPL increased and the average EP decreased after operation. After operation the average G decreased from 2.8 to 1.7 (P < 0.01) and the percentage of improvement over one grade was 71%. CONCLUSION: From these results mentioned above, it can be concluded that the arytenoid adduction operation might be one of the most effective operative methods to improve glottal incompetence caused by laryngeal nerve paralysis. PMID- 12541868 TI - [Some problem in diagnosis and treatment of closed laryngeal trauma]. AB - OBJECTIVE: Raise the diagnosis and treatment level of laryngeal closed trauma. METHOD: Analyze and discuss some problems of diagnosis or treatment through five cases reports. RESULT: Except clinical signs and symptoms, fibrolaryngoscopy and CT or MRI imaging examination should be used. According to it, make the injury degree of intralaryngeal structure and decide the management principle. CONCLUSION: Corrective diagnosis and treatment in emergency period was the key to raise curative effect in the near future and to prevent laryngeal stenosis in late period. PMID- 12541869 TI - [Prognostic significance of CD57+ cell level and pTNM staying system in patients with laryngeal cancer]. AB - OBJECTIVE: To analyze the prognostic value of the pTNM clinical staying system, as well as the CD57+ cells level in peripheral blood samples of the patients with laryngeal carcinoma. METHOD: The CD57+ cells level of peripheral blood was determined with SAP method in 34 patients with laryngeal carcinoma and 16 normal subjects. The pTNM clinical stage was T1-2 in 13, T3-4 in 21; pN+ in 10, pN0 in 24; supraglottis in 8, glottis in 23; hypoglottis in 3; pathologic differentiation G1 in 11, G2,3 in 23. RESULT: The CD57+ cells level significantly increased (pN+ vs. pNo). In the 12nd month actual survival were significantly different (T1,2 vs. T3,4, pN0 vs. pN+, G1 vs G2,3, supraglottis vs. glottis and hypoglottis, CD57+ > or = 15% vs. CD57+ < 15%). In the 24th month and 36th month actual survival were significantly different (pN0 vs. pN+, G1 vs G2,3, supraglottis. glottis and hypoglottis, CD57+ > or = 15% vs. CD57+ < 15%). In the 48th month actual survival were significantly different only (pN0 vs. pN+, supraglottis vs. glottis and hypo glottis, CD57+ > or = 15% vs. CD57+ < 15%). CONCLUSION: These results suggested that the assessment of pTNM system and the CD57+ cells level have significance for prognosis of patients with laryngeal carcinoma. PMID- 12541870 TI - [Study on susceptibility test of pathogenic fungi from otorhinolaryngology by Etest method]. AB - OBJECTIVE: To study the sensitivity of pathogenic fungi from otorhinolaryngology to 5 antifungal agents of Ketoconazole (KE), Itraconazole(IT), Fluorocytosine(FC), Amphtericin B (AP) and Fluconazole(FL) by Etest method. METHOD: Etest for determine MIC of 14 pathogenic fungi was performed according to the manufacturer's instructions. The Etest strip containing the 5 antifungal drugs were putted on medium with Candida albicans, A. flarus and A. fumigatus etc., respectively, to determine MIC of antifungal drugs. RESULT: Candida albicans were sensitized to all the above 5 antifungal drugs, KE and IT were most sensitized. Seven fungi such as A. flavus, A fumigatus, A. oryzae etc were all sensitized to KE and IT. The MIC range of KE and IT against 15 strains of pathogenic fungi were < or = 0.008-2 mg/L and < or = 0.006-4 mg/L respectively. In 15 strains, 11 strains showed drug-resistant to FL, 8 strains to AP and FC, 4 strains to KE and 1 strains to IT respectively and MIC were all > 32 mg/L. CONCLUSION: Etest MIC were in good agreement with macrobroth MIC. The use of Etest strips for antifungal susceptibility test is a new and promising method with advantage of easy to perform, exact results and satisfactory reproducibility. Etest is valuable on clinical practice. PMID- 12541871 TI - [Coefficients between TEOAEs and DPOAEs in infants]. AB - OBJECTIVE: To investigate the coefficients of TEOAEs and DPOAEs based on various pass/fail criteria. METHOD: Thirty-six Australia infants were tested (age range: 1-6 months) using both TEOAE and DPOAE. Cohen's Kappa (K) were used to analyze different criteria. RESULT: There is a significant low agreement in the screening outcomes between TEOAEs and DPOAEs, and different criteria for TEOAEs. CONCLUSION: The findings from the study of coefficients among commonly used screening pass/fail criteria suggested that there is no simple relationship among them. The possible reasons for weak agreement were discussed. PMID- 12541872 TI - [Primary restoration for congenital malformation of external and middle ear]. AB - OBJECTIVE: To promote the hearing, improve the appearance, shorten the therapeutic course in congenital malformation of external and middle ear, the program of primary restoration has been designed. METHOD: Under general or local anaesthesia a Z-shaped incision is made in skin. After turn up the flap A (defective ear) and flap B(retroauricular skin), a periosteal flap C with pedicle in front is made and elevated from mastoid. Then made frameworklize on mastoid and perform tympanoplasty. Once the myringograft is put properly the flap C is turn into the mastoidal cavity to serve as the lining of anterior wall. Flap A is sutured with the edge of retroauricular incision to serve as the lower part of new auricle. Then use the flap B to wrap a siliceous frame to make the upper part of new auricle. Finally, the naked walls of mastoidal cavity are lined with free skin graft and the cavity is packed with vaseline gauze for two weeks. RESULT: Two cases of congenital malformation of external and middle ear were treated with the procedure mentioned above and were followed up for one and 3 years respectively. Both effects of shape and hearing were good. CONCLUSION: The primary restoration for congenital malformation of external and middle ear designed by the authors is an excellent method worthy to be recommended. PMID- 12541873 TI - [Diagnostic significance of audiological and vestibular function examination in the pontocerebellar angle tumor]. AB - OBJECTIVE: To study the diagnostic value of audiological and vestibular function examination in the pontocerebellar angle tumor. METHOD: The data of audiological and vestibular function test were analized retrospectively in 20 subjects with tumors of the pontocerebellar angle. RESULT: Pure tone and ABR waveforms were abnormal in 20 subjects (21 ears), moreover acoustic stapedius reflex and the vestibular function were abnormal in 19 subjects. CONCLUSION: It's valuable that audiological and vestibular function examinations are able to diagnosis the pontocerebellar angle tumor. PMID- 12541874 TI - [The study of TNF-alpha destructive action to the bone in human middle cholesteatoma]. AB - OBJECTIVE: To study the expression of TNF-alpha in the cholesteatoma and elucidate the role of TNF-alpha in the destruction of bone. METHOD: All samples (5 microns paraffin sections) from 25 cholesteatoma cases and 10 normal cases were examined by immunohistochemical SABC method and analysed by computer image. RESULT: In 25 cholesteatoma cases, TNF-alpha expressed in the cytoplasm of the full epithelial tissue cells, subcutaneous inflammatory cells and fibroblast. In contrast, the normal cases, there were 6 cases week positive, 4 cases negative. The results of the computer image quantitative analysis system showed that the mean optical density of TNF-alpha was 0.1326 +/- 0.0022 in the cholesteatoma epithelial tissue and 0.0868 +/- 0.0014 in normal skin epithelial tissue respectively, both of which had significantly differentiation (P < 0.05). CONCLUSION: TNF-alpha may be a factor of the destruction of bone of cholesteatoma. PMID- 12541875 TI - [Appraisement of the operative therapeutical effects of salpingocatheterism for refractory otitis media]. AB - OBJECTIVE: The therapeutical effects of salpingocatheterism and the states of tubal function were studied after different middle ear operations and within different middle ear pathological changes. METHOD: Total 71 patients with tubal occlusion, 18 of myringoplasty, 20 of reconstruction of ossicolar chain followed tympanoplasty, 14 of combined approach tympanoplasty, 10 of radical mastoidectomy followed tympanoplasty, 9 of re-radical mastoidectomy followed tympanoplasty were implemented salpingocatheterism. The patients were followed up for evaluation of the postoperative therapeutical effects and the state of tubal function. RESULT: The causes of operation failure in different middle ear operations or different middle ear pathological changes had no significantly difference. The major cause for postoperative tympanic membrane perforation and tubal occlusion was the pathogenic damage being not cleared up thoroughly and/or there being nasal cavity or pharyngeal cavity pathogenic changes accompanied. CONCLUSION: Salpingocatheterism is effective for preventing eustachian tube postoperative adhesion. PMID- 12541876 TI - [Reconstruction of post traumatic auricle partial defect by post-auricle flap schlepping muscle root]. AB - OBJECTIVE: To study the method of repairing post-traumatic auricle partial defect. METHOD: 11 cases of post-traumatic auricle partial defect had been repaired by post-auricle flap schlepping muscle root in our hospital, the follow up period from 3 months to 1 year. RESULT: It shows excellent shape. The flap necrosis hadn't been happened. CONCLUSION: Advantages of this procedure were simple to manage, texture match and good skin color. The donor scar was unconscious. It is a choice for treating post-traumatic auricle partial defect. PMID- 12541877 TI - [Experiences of surgical treatment with cicatricial laryngotracheal atresia: report of 48 cases]. AB - OBJECTIVE: In order to improve the treatment methods of laryngotracheal atresia, several improved surgical procedures were used. METHOD: From 1986, the 48 patients with laryngotracheal atresia accepted surgical treatment in our hospital. Laryngo-fissure with laser cauterization of the glottis, laryngotracheal reconstruction using surgical flaps with or without particles, laryngo-tracheal reconstruction using itself's cicatrices, tracheal end-to-end anastomosis, two-stage laryngotracheal reconstruction using aqueduct method, bronchotracheal reconstruction through the chest. RESULT: 46/48 cases were cured breathing without tracheal cannula. Following-up survey is 1-12 years. CONCLUSION: The treatment of cicatricial laryngotracheal atresia was difficult, but if the surgical methods were selected carefully according to the situation of each patient, we also can get a good result. PMID- 12541878 TI - [The relationship between acoustic analysis and auditory assessment]. AB - OBJECTIVE: To investigate the relationship between acoustic analysis and auditory assessment of hoarseness. METHOD: 24 normal and 154 cases of pathological voice were included in this research. They were divided into 4 groups with different hoarseness and were accepted acoustic analysis. The parameters were analyzed by SPSS statistical software. RESULT: There were significant difference on acoustical parameters among each group excluding no difference of the jitter between normal and light hoarseness. NNE and SNR were the major parameters in descriptive of hoarseness. There were good correlation between 5 parameters and hoarseness assessment. CONCLUSION: Acoustic analysis is a useful method in clinical application to assess hoarseness with objective and simple advantage. PMID- 12541879 TI - [Local microcirculatory disorders in the cochlea basal turn near termination in guinea pigs]. AB - OBJECTIVE: To study the characteristics of hearing loss caused by local microcirculatory disorders in the cochlea basal turn near termination of guinea pigs. METHOD: The local microcirculatory disorders in the cochlea basal turn near termination of guinea pigs were caused by photochemical reaction; morphological changes were observed by light microscope; the CAP N1 amplitude, CAP N1 latency and CAP N1 threshold shift which were evoked by short tone burst were recorded by Madsen 2250. RESULT: Morphological changes showed that histopathological changes were located in the basal turn near termination of cochlea. The CAP N1 amplitude, CAP N1 latency and CAP N1 threshold shift showed that the hearing loss was remarkable in the high-range frequencies. CONCLUSION: If the local microcirculatory disorders in the cochlea basal turn near termination take place, the hearing losses are more remarkable in the high-range frequencies. PMID- 12541880 TI - [Isolation of outer hair cells from varying turns of the guinea-pig cochlea]. AB - OBJECTIVE: To probe the method of isolating outer hair cells (OHC) from each of four turns of the guinea-pig cochlea. METHOD: From eight guinea pigs the organ of Corti from each of four turns of the cochlea were dissected, and then treated using enzyme. RESULT: A fair amount of living OHCs from each of four turns were obtained. The length of OHCs from each of four turns were 23.81, 34.50, 60.48 and 71.37 microns. CONCLUSION: The key to success in isolating OHCs from each of four turns of the cochlea is to know very well the anatomical characteristics of each of four turns of the cochlea and be operated in accordance with normal rules. PMID- 12541881 TI - [Posterior semicircular canal occlusion with microwave assisted technique in guinea pigs]. AB - OBJECTIVE: To study the effectiveness of posterior semicircular canal occlusion with microwave assisted technique. METHOD: Posterior semicircular canal occlusion with microwave assisted technique was performed in 20 guinea pigs. Electronystagmography (ENG) and auditory brainstem response (ABR) were recorded preoperatively and postoperatively. Morphological changes were observed under light microscopy. RESULT: Left and right nystagmus returned symmetrical on the 3rd postoperative day. Two ears showed normal symmetrical nystagmus to icc-water stimulation on preoperation and the 30th postoperative day. There were no significant changes of the thresholds of ABR before and after surgery. Histopathological findings revealed that posterior semicircular canal was completely occluded, and that Organ of Corti and crista ampullaris were normal. CONCLUSION: Posterior semicircular occlusion with microwave technique is suggested as an effective and safe treatment for the benign paroxysmal positional vertigo. PMID- 12541882 TI - [The clinical and pathological research of supraglottic carcinoma with bilateral cervical nodal metastasis]. AB - OBJECTIVE: To evaluate clinical and pathological characteristics in patients with supraglottic carcinoma and bilateral cervical nodal metastasis. METHOD: 60 patients with supraglottic carcinoma treated in this hospital from 1985 to 1994 were retrospectively reviewed, including 30 cases with bilateral metastasis (experiment group) and 30 cases with unilateral metastasis (control group). The two groups were contrasted according to site of tumor, tumor diameter, surface morphology, clinical staging, the degree of invading the contralateral semilarynx, Border's grading, growth mode of tumor edge, the degree of invading preepiglottic space. RESULT: In stage T4 cases with low grading of cell differentiation made up 63.6% (7/11) in the bilateral metastasis group, while 10% in the ipsilateral metastasis group (P < 0.05). Significant differences lied between N0 group and N2 group (P < 0.05). Moderate and severe invasion of contralateral semilarynx in the bilateral group made up 87.5% (14/16), significantly higher than that in ipsilateral group 53.3% (16/30). CONCLUSION: Bilateral neck dissection should be used in T4 cases with low grading of cell differentiation in which the contralateral semilarynx was invaded. Bilateral neck dissection should be taken for the treatment of supraglottic carcinoma with N2.3 diseases. Bilateral neck dissection is recommended for the patients with moderate and severe invasion of the contralateral semilarynx. PMID- 12541883 TI - [Substituting esophagus with colon in the treatment of hypopharyngeal and esophageal disease]. AB - OBJECTIVE: To study the feasibility and effect of substituting esophagus with colon in the treatment of advanced stage hypopharyngeal carcinoma, cervical esophagus cancer and serious esophageal stenosis on the basis of laryngeal function preservation. METHODS: From 1989 to 1996. 25 patients in our department were retrospectively reviewed among them, nine were hypopharyngeal carcinoma, T3 (No1, N1 4, N2 2), T4 N1 M0 2 (UICC 1997), thirteen were cervical esophagus cancer, T1N0M0 2, T2 (N0 4, N1 7), there were serious esophageal stenosis, length was 3-5 cm. The patients with carcinoma were given postoperation radiotherapy. RESULT: The total survival rates in 3, 5 years are 54.5%, 42.9% respectively. The 3-year survival rates are 44.4% in the cases with advanced stage hypopharyngeal carcinoma, and 61.5% with cervical esophagus cancer, respectively. CONCLUSION: Substituting esophagus with colon is in conformity with physical demand, can reduce complication and improve life quality. PMID- 12541884 TI - [The study on the occult metastases to cervical lymph node in squamous cell carcinoma of the larynx and hypopharynx]. AB - OBJECTIVE: To study the metastastic regularity of cervical lymph node in squamous cell carcinoma of the larynx and hypopharynx, which will provide some theoretical basis for surgeon to use selective neck dissection (SND) in some patients with carcinoma above. METHOD: Sixty-nine originally cN0 staged neck dissection specimens from 40 patients were evaluated using mutipl-level sectioning in 4 microns intervals, which were respectively processed by H&E-staining and immunostaining with an antibody to cytokeratin (CK AE3). These patients weren't given any treatment (radiotherapy or chemotherapy or surgery in neck) before surgery. All cases had been followed up 1 year or more. RESULT: The examination of lymph nodes from 40 patients revealed 31 (0.59% 31/2219) occult metastases in 14 (35% 14/40) patients. Among them, six patients with supraglottic cancer, 1 patient with transglottic cancer and 7 patients with hypopharyngeal cancer. There were no occult metastases to cervical lymph node among 9 patients with glottic cancer. All occult metastases mainly located in ipsilateral level II and/or level III. Occult metastases weren't found in nodes from level I, level IV and level V. CONCLUSION: The study further prove that metastases to cervical lymph node mainly occur in the ipsilateral level II and level III among patients with carcinoma of the larynx and hypopharynx. According to the risks of occurring occult metastases to cervical lymph node, we suggest applying to ipsilateral or bilateral SND (level II and level III be dissected) among patients with carcinoma of the larynx and hypopharynx which staged T2-T4 N0M0 clinically. PMID- 12541885 TI - [Diagnosis and surgical treatment of carotid body tumor]. AB - OBJECTIVE: To promote the rate of diagnosis and successful rate of operation for carotid body tumor. METHOD: 26 cases of the 30 patients with carotid body tumor were performed by DSA and 5 cases of them, feeding arteries of tumor were embolized with gelfoam through superlective catheterization, preoperativelly. The tumor was resected under carotid adventitial plane in 12 cases, with extracervical artery in 8 cases and with tumor-carotid segment block in 10 cases. RESULT: Of the 30 cases, no one died from operation or cerebral vascular complications. DSA plays a significant role in diagnosis of the tumor. Intraoperative bleeding can be remarkably reduced if feeding arteries were embolized preoperativelly. The tumor fixed to segment of the carotid arteries block was resected and the reconstruction of vasculature was unnecessary as soon as candidates were certified to be qualified by carotid training and DSA or/and TCD tests. Follow up of these 30 patients revealed that 25 of them survived free from cerebrovascular complications and tumor recurrence for 5 months-10 years. 1 of them died from traffic accident 2 year after operation, 4 of them had lost communication. CONCLUSION: DSA is a most authritative approach for diagnosis of carotid body tumor, DSA and TCD tests play a key role to resect carotid artery without reconstruction. Successful operation based on a precise preoperative evaluation, choice of operative ways and proper procedure. PMID- 12541886 TI - [Cricohyoidoepiglottopexy: a report of 21 cases]. AB - OBJECTIVE: To assess the indications, function preserving and long term results of cricohyoidoepiglottopexy (CHEP) in the treatment of laryngeal carcinoma. METHOD: Twenty-one patients were underwent this procedure during the period from 1993 to 1999 in our department. There were 20 glottic cancers (7 T1N0M0, 10 T2N0M0, 3 T3N0M0) and 1 supraglottic type (T2N1M0) according to the 1997 UICC system. In these cases, 15 were the aged over 60 years. RESULT: First decannulation rate was 95.24%, and majority completed in 28 days post-operation. No patient was with cannula all his life. All cases could finally take food by mouth without aspiration. Speech was also satisfactory. The overall 3-year and 5 year survival rates were 87.50% and 75.00% respectively. No patient experienced local recurrence. CONCLUSION: This procedure is presented as a useful alternative to partial vertical laryngectomy and total laryngectomy in select cases of laryngeal carcinoma. PMID- 12541887 TI - [Subacute thyroiditis as seen initially in otorhinolaryngology department: a report of 30 cases]. AB - OBJECTIVE: To assess the pharyngeal presentations and the diagnostic value of thyroid SPECT and thyroid fine needle biopsy (FNB) in subacute thyroiditis (SAT) as seen initially in otorhinolaryngology department. METHODS: Thirty patients with SAT were mainly recorded about pharyngeal symptoms and performed about serum T3, T4 level and thyroid SPECT imaging or combining thyroid FNB. RESULT: Of 30 patients, 21 had variant sore throat, 9 had abnormal sensation of throat. 6 were diagnosed as having SAT by only SPECT, the remaining 24 by that combined with FNB, 2 of whom were finally considered as having SAT by oral Prednisone therapy. CONCLUSION: Sore throat and abnormal sensation of throat are the important presentations of SAT, thyroid SPECT imaging and thyroid FNB are valuable on diagnosing SAT. PMID- 12541888 TI - [The correlation of multidrug resistance phenotype with clinical response to chemotherapy in laryngeal cancer]. AB - OBJECTIVE: The correlation of multidrug resistance(MDR) phenotype with clinical response to chemotherapy was investigated in patients with laryngeal cancer. METHOD: Tumor specimens prior to neoadjuvant chemotherapy from 36 cases of laryngeal cancer were collected for detection of P-glycoprotein (P-gp) with anti Pgp monoclonal antibodies (JSB-I). Immunohistochemical assays were used for P-gp detection on 5 microns thick frozen section. Anti-Pgp monoclonal antibodies(JSB I) as primary antibodies and goat anti-mouse IgG monoclonal antibodies as second antibodies were applied. All patients received preoperative induction chemotherapy with a regimen of cisplatin, 5-fluorouracil and pingyangmycin. Evaluation for tumor response was scored as follows: Response to chemotherapy was graded as good for complete response or partial response; as poor for stable disease or progressive disease. RESULT: Of the 36 pretreatment specimens, 12(33.3%) were P-gp positive and 24 (66.7%) were P-gp negative. Among the 12 patients with P-gp positive, 3(25%) had a good response and 9(75%) had a poor response. Of the 24 patients with P-gp negative, 21(87.5%) had a good response and 3(12.5%) had a poor response (P < 0.05). CONCLUSION: These preliminary data suggest that a significant correlation between MDR phenotype, Pgp-mediated and chemotherapy resistance existed. PMID- 12541889 TI - [Application of CT virtual bronchoscope in foreign-body in children's bronchus]. AB - OBJECTIVE: To evaluate the clinical application of CT virtual bronchoscope in foreign-body in bonchus. METHOD: The CT virtual bronchoscope were used in 21 cases with foreign-body in bronchus, all the images and reports were compared with the actual bronchoscopy. RESULT: The reports and images of CT virtual bronchoscope are very agreed with that of actual bronchoscopy. CONCLUSION: CT virtual bronchoscope is a very good complemental method in preoperative diagnosis of foreign-body in bronchus. PMID- 12541890 TI - [Comparison of the post-operative complication in head-neck malignant neoplasms between aged and pre-aged]. AB - OBJECTIVE: To compare the post-operation complication of head-neck malignant tumor in the aged with pre-aged. METHOD: 80 aged and 92 pre-aged cases of head neck tumor were performed operation. In the aged group, 26 cases were performed with preoperative radiotherapy and 13 cases with postoperative radiotherapy. In the pre-aged group, 40 cases were performed with preoperative radiotherapy and 6 cases with postoperative radiotherapy. RESULT: The incidence of complication in the aged is higher than that in the pre-aged group. Moreover, the post-surgery complications of tumor in head and neck in the aged preferably appear in the cardiopulmones and urology. In contrast, most of the complication in the pre-aged appears in the incision of operation. CONCLUSION: Prior to surgery, we should clear with the situation of patients, and consequently choose appropriate methods of operation. Age does not play an essential role in the decision of the methods of operation. PMID- 12541891 TI - [Improved retrosigmoid approach operation in the treatment of trigeminal neuralgia]. AB - OBJECTIVE: To explore the surgical method which can raise the treatment effect and reduce the complication occurrence of cerebellopontine angle (CPA) operation by retrosigmoid approach. METHOD: Have improved the retrosigmoid approach in the skin incision, the hole position of bone, the intracranial operation skills and the duramatral suture, etc. By the improved retrosigmoid approach, we used partial sensory rhizotomy (PSR) to cure 54 cases with trigeminal neuralgia (TN). RESULT: 52 cases got satisfactory treatment effect and no complication appeared. In the other 2 cases, the trigeminal nerve was not found in CPA. CONCLUSION: The CPA operation by retrosigmoid approach is dangerous relatively, it usually can cause some serious complication. But improved retrosigmoid approach can expose CPA sufficiently and is convenient to operate. On the other hand, it can decrease intracranial unexpected hemorrhage and complication occurrence. PMID- 12541892 TI - [Application of nasal endoscope in diagnosis and treatment of epistaxis]. AB - OBJECTIVE: To investigate the indication for diagnosis and treatment of epistaxis under nasal endoscope. METHOD: The data of 226 epistaxis cases were analized. Under nasal endoscope: 126 cases were packed, 78 cases were treated with microwave, laser cautery, 42 cases were operated. RESULT: An outstanding visutization was afforded by nasal endoscope and operation conveniencly, diagnosis correctively. CONCLUSION: We think it is an excellent method to examine and treat epistaxis and worthy to be recommented. PMID- 12541893 TI - [Measurement of distortion product otoacoustic emissions in guinea pig]. AB - OBJECTIVE: To establish a stable detecting method of distortion product otoacoustic emissions (DPOAEs). METHOD: After cutting off tragus, seven healthy pigmented guinea pigs (thirteen ears) were selected to measure the DPOAEs at the frequency of 2f1-f2 (cubic DPOAEs). The I/O function and latency at f2 = 2, 4, 6 kHz were analyzed. RESULT: The incidence of DPOAEs at all frequencies was 100%. The maximal amplitude of input-output function was (27.24 +/- 4.15) dB SPL, (27.98 +/- 4.05) dB SPL and (41.45 +/- 1.20) dB SPL respectively; latency was (2.06 +/- 0.21) ms, (1.52 +/- 0.16) ms and (1.37 +/- 0.08) ms at the corresponding frequencies. CONCLUSION: After the improvement of detection, it is satisfactory to measuring DPOAE in animals because of the high incidence, high amplitude and good repetition. PMID- 12541894 TI - [Expression and the role of vascular endothelial growth factor mRNA in nasal polyps]. AB - OBJECTIVE: To explore the pathogenesis of nasal polyps, the aim of this study is to detect the expression of vascular endothelial growth factor (VEGF) in nasal polyp(NP) tissue, and compare these findings with normal nasal turbinate. METHOD: We examined the expression of VEGF in human nasal polyps from patients underwent nasal polypectomy (n = 7) and nasal turbinate from patients with snoring (n = 6). VEGF mRNA expression was determined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULT: VEGF mRNA expression was significantly stronger in the eight NP than in the turbinate tissue group (P < 0.05). CONCLUSION: We conclude that VEGF expression is upregulated in NP, which suggests a potential role of VEGF in the formation and growth and recrudescence of NP. PMID- 12541895 TI - [Research of drug resistance and detection of methicillin-resistant Staphylococcus aureus in chronic maxillary sinusitis]. AB - OBJECTIVE: To investigate the detection method of methicillin-resistant staphylococcus aureus (MRSA) in chronic maxillary sinusitis and the situation of drug resistance to some common antibiotics. METHOD: Using multiplex polymerase chain reaction technique to detect the drug resistant gene (mecA) of MRSA and specific gene (femA) of staphylococcus aureus at the same time. Drug resistant tests were performed with disk diffusion (K-B) method. RESULT: femA is a specific gene of staphylococcus aureus, the detective ratio of mecA in MRSA is 96.2% (51/53). Vancomycin is the only antibiotic which is sensitive to MRSA, while others are not. CONCLUSION: MRSA can be rapidly and specifically identified with multiplex PCR. There are drug resistance to many antibiotic drugs for MRSA. PMID- 12541896 TI - [Revision endoscopic sinus surgery on chronic sinusitis and polyps]. AB - OBJECTIVE: To study the indications, surgical landmarks, surgical significance, complications and operative avoidance of revision endoscopic sinus surgery on chronic sinusitis and polyps. METHOD: We performed revision endoscopic sinus surgery on 112 cases of chronic sinusitis and polyps under controlled hypotension anaesthesia by using the Messerklinger technique and Stryker debrider. RESULT: 89 cases were successful and 17 unresolved, 6 failed, all the complications consist of cerebrospinal fistula in one, orbital hematoma in four, increased tearing in three and profuse bleeding in 6 patients. CONCLUSION: Revision endoscopic sinus surgery was advised in case of sinuous obstruction caused by uncontrolled lesions under postoperative care, which can do good for adhesion and obstruction except nasal polyposis. The remanet of the middle turbinate and the upper arch of choana were helpful operative landmarks. Preoperative CT scan decrease the complications. PMID- 12541897 TI - [Diagnostic value of computer-based endoscopy system]. AB - OBJECTIVE: To assess the roles of computer-based endoscopy system as a clinical tool in evaluation of laryngeal disorder. METHOD: An extensive research of 100 patients under stroboscopy examination with acoustic and EGG signal was made, and a number of quantitative information, such as contact quotient, glottal noise and glottal area changes, was examined. RESULT: Quantitative information contributes significant diagnostic information in 14 cases, resulted in avoidance of surgery in 6 cases and 8 cases referred to surgery instead of voice and medical therapy. CONCLUSION: Acoustic and EGG measures should be used as substantially complements of endoscopy examination in the routine clinical examination. PMID- 12541898 TI - [Relationship between operative time and therapeutic effectiveness after the optic nerve injuries]. AB - OBJECTIVE: To understand the relationship between the operative opportunity and effectiveness in the optic nerve injuries. METHOD: Twenty-three cases with optic nerve injuries were undertaken optic canal decompression. RESULT: The visual acuity in 13 cases were improved after the operation, and progressed about 0.233 averagely. The average time from injuries to operations was (12.90 +/- 16.77) d (3-22 d) in ineffective group. 7 cases of hypoactivity of eyeball recovered in 8 cases. CONCLUSION: The optic nerve decompression should be done after the patients were treated with megadose of steroid for 48 hours and did not recover. The time had better be no more than 7 days. PMID- 12541899 TI - [The clinic treatment of traumatic optic neuropathy]. AB - OBJECTIVE: To observe the visual outcome of traumatic optic neuropathy treated with optic canal decompression surgery and medication. METHOD: 15 patients, suffering from traumatic optic neuropathy, had received medication and optic canal decompression surgery through the approach of ethmoid and sphenoid sinus. The vision acuity was divided into 5 grades: no light perception, light perception, finger movement, counting fingers, vision > or = 0.1, the vision acuity improvements reaching 2 grades or more were determined as effective. All patients were followed up for intervals of 1 month and 6 months. RESULT: The total efficient rate is 57.4%, no intraoperative or postoperative complications were encountered. Being followed up 6 months, 2 cases, who underwent respectively the operation 8, 11 days after trauma, felt no light perception. CONCLUSION: The approach through ethmoid and sphenoid sinus have the merits such as safety, less damage and larger operative field. Timely medication, megacorticosteroid therapy mainly, may play a good role in alleviating optic nerve edema and restoring its function. Comprehensive therapy may be an ideal treatment to traumatic optic neuropathy. PMID- 12541900 TI - [Plasma cell granuloma of maxillary sinuses]. AB - OBJECTIVE: To study the clinical features and treatment of plasma cell granuloma in maxillary sinuses. METHOD: The clinical information of 3 cases was reported and the literatures were also reviewed. RESULT: All of the 3 cases were cured after Caldwell-Luc operation combined with oral corticosteroids and antiallergic agent. Recurrence hasn't been found in half to 2 years follow-up period. CONCLUSION: Plasma cell granuloma of maxillary sinus is a chronic inflammatory lesion with unknown origin. In almost all of the cases, CT images showed some degree of involvement of the bony wall of the maxillary sinuses. The imaging findings mimicker malignant tumors. But the prognosis were quite well in most of the patients. PMID- 12541901 TI - [The effect of submucosal diathermy to the inferior turbinates on nasal airway resistance]. AB - OBJECTIVE: The main purpose of the study is to observe the effect of submucosal diathermy to the inferior turbinates on nasal airway resistance (NAR). METHOD: Using active anterior rhinomanometry, NAR were measured in seventeen patients who suffered from chronic rhinitis before and post operation. The subjective impression of their nasal patency was assessed using a 10-point scale. RESULT: The NAR is(0.662 +/- 0.22) kPa.s/L before operation and (0.302 +/- 0.17) kPa.s/L three months after operation. The NAR is 0.397 +/- 0.25 ten months after operation. The differences are statistically significant (P < 0.01) between the NAR before and post operation. There are significant differences in the subjective impression of patients' nasal patency as well (P < 0.01). CONCLUSION: Submucosal diathermy to the inferior turbinates is effective in decreasing NAR and improving the subjective impression of their nasal patency. PMID- 12541902 TI - [The clinical value of the TSGF pre and post-therapy in patients with nasopharyngeal carcinoma(NPC)]. AB - OBJECTIVE: To investigate the role of TSGF (tumor supplied group of factor) in early diagnosing and predicting in patients with NPC. METHOD: EBV-IgA and level of TSGF were measured before therapy and 1 month, 1 year after therapy in 35 patients with NPC, and the relationship between them was researched. RESULT: The diagnostic sensitivity of TSGF and EBV-IgA was 91%, 97% respectively, there is no significant relation between them. After treatment, the level of TSGF in PD (progression disease) patient remained higher, but had a significantly drop in RR(including the CR, PR and SI)) patient. CONCLUSION: Results suggested that TSGF may be useful in diagnosing and prediction of NPC. PMID- 12541903 TI - [Clinical analysis of 17 cases rhabdomyosarcoma in ear nose and throat]. AB - OBJECTIVE: To better understand the rhabdomyosarcoma in ear nose and throat and make correct diagnosis. METHOD: 17 patients with rhabdomyosarcoma in ear nose and throat were investigated. Diagnosis, treatment and prognosis were analyzed retrospectively. RESULT: Among 17 patients with rhabdomyosarcoma, three patients were in stage I, four patients were in stage II, eight patients were in stage III and two patients were in stage IV. It is the commonest in nose and predisposed to male. The clinic presentations were related to the size and location of tumor. Comprehensive therapy was need in most of patients. Poor prognosis was noted in this disease. CONCLUSION: This disease is easy to misdiagnose. Early diagnosis and comprehensive therapy will make contribution to better prognosis. PMID- 12541905 TI - [The application of mastoid cavity plombage in open method tympanoplasty]. AB - OBJECTIVE: The role mastoid cavity plombage played in restoring physiological function of external meatus in open method tympanoplasty was studied. METHOD: 85 cases of cholesteatoma otitis media were performed open method tympanoplasty with the broken bits of their own mastoid process cortex, homological costal cartilage or U shape periosteum connective tissue valve. And plombage was performed at the same time. The reconstruction of ossicular chain (type 0 tympanoplasty): 10 ears did not undergo reconstruction due to increase ment of bone conduction threshold or were in preparation for the second phase operation; 40 ears underwent type III tympanoplasty; 35 ears underwent type IV tympanoplasty. RESULT: The total improvement rate of hearing was 76%, among which 80% were performed type III tympanoplasty and 71.4% underwent type IV tympanoplasty. After operation, the time waited till dry-ears was 19.8 +/- 65.31 days and the dry-ears rate was 97.6% (83/85). Six months after operation, 89.4% (76/85) had smooth external meatus and fine self-cleaning function and no recurrence was found. CONCLUSION: As to cholesteatoma otitis media that could not be performed closed method tympanoplasty, U shape periosteum connective tissue valve and mastoid process cortex were used. Mastoid cavity plombage with homological costal cartilage could effectively overcome the shortcomings of destruction of physiological structure of external meatus caused by open method tympanoplasty, the self-cleaning function of skins of external meatus was remained and the life quality of patients after operation was increased. PMID- 12541904 TI - [Detection of telomerase activity in laryngeal carcinoma and the association between telomerase activity and lymphatic metastasis]. AB - OBJECTIVE: To investigate the telomerase activity in human laryngeal squamous cell carcinomas (LSCC) and the association between telomerase activity and lymphatic metastasis. METHOD: Telomerase activity was determined in 47 specimens of LSCC and 10 specimens of laryngeal inflammatory polyps using PCR-ELISA. RESULT: Telomerase activity was detected in 39 of 47 (83%) LSCC, but was not detected in all 10 laryngeal inflammatory polyps. There were significant differences between them (P < 0.01). Telomerase activation in LSCC with positive cervical lymph node involvement appeared to be higher (13 out of 13 cases, 100%) than in LSCC with negative cervical lymph node involvement (26 out of 34, 76.47%), (P < 0.05). CONCLUSION: Telomerase is activated frequently in LSCC and may become a biomarker of LSCC. The detection of telomerase activity in LSCC may have some value for predicting metastasis potential and prognosis and can guide the clinical practices applying selective neck dissection. PMID- 12541906 TI - [Effect of the Purariae-isofiavones on atrophic nasal mucosas in ovariectomized rats]. AB - OBJECTIVE: To study the effect of Purariae-isoflavone on atrophic nasal mucosas in ovariectomized rats. METHOD: 60 rats were divided into four groups randomly, every group with 15 rats. The first group was studied as normal control group. Rats of the second group were ovariectomized and not given any treatment. Rats of the third group were given Weinian by gastrogavage from 30 days after being ovariectomized to being killed. Rats of the forth group were given purariae isoflavone by gastrogavage from 30 days after operation to being killed. Level of Estrodiol (E2) of every rat was measured. Morphological changes of the nasal mucosas were observed by light-microscopy and electron-microscopy. RESULT: 30 days after being ovariectomized, atrophic nasal mucosas occured, and average E2 level was lower significantly than that of control group (P < 0.05). Atrophic mucosas recovered after being treated with Weinian or Purariae isoflavone. E2 level of the third group was lower significantly than that of the first group and the second group (P < 0.05). E2 level of the forth group recovered (P > 0.05). CONCLUSION: Purariae-isoflavone have effects on treating the atrophic nasal mucosas and recovering E2 level in ovariectomized rats. PMID- 12541907 TI - [Fast optimization of stepwise gradient conditions for ternary mobile phase in reversed-phase high performance liquid chromatography]. AB - In high performance liquid chromatography, it is necessary to apply multi composition gradient elution for the separation of complex samples such as environmental and biological samples. Multivariate stepwise gradient elution is one of the most efficient elution modes, because it combines the high selectivity of multi-composition mobile phase and shorter analysis time of gradient elution. In practical separations, the separation selectivity of samples can be effectively adjusted by using ternary mobile phase. For the optimization of these parameters, the retention equation of samples must be obtained at first. Traditionally, several isocratic experiments are used to get the retention equation of solute. However, it is time consuming especially for the separation of complex samples with a wide range of polarity. A new method for the fast optimization of ternary stepwise gradient elution was proposed based on the migration rule of solute in column. First, the coefficients of retention equation of solute are obtained by running several linear gradient experiments, then the optimal separation conditions are searched according to the hierarchical chromatography response function which acts as the optimization criterion. For each kind of organic modifier, two initial linear gradient experiments are used to obtain the primary coefficients of retention equation of each solute. For ternary mobile phase, only four linear gradient runs are needed to get the coefficients of retention equation. Then the retention times of solutes under arbitrary mobile phase composition can be predicted. The initial optimal mobile phase composition is obtained by resolution mapping for all of the solutes. A hierarchical chromatography response function is used to evaluate the separation efficiencies and search the optimal elution conditions. In subsequent optimization, the migrating distance of solute in the column is considered to decide the mobile phase composition and sustaining time of the latter steps until all the solutes are eluted out. Thus the first stepwise gradient elution conditions are predicted. If the resolution of samples under the predicted optimal separation conditions is satisfactory, the optimization procedure is stopped; otherwise, the coefficients of retention equation are adjusted according to the experimental results under the previously predicted elution conditions. Then the new stepwise gradient elution conditions are predicted repeatedly until satisfactory resolution is obtained. Normally, the satisfactory separation conditions can be found only after six experiments by using the proposed method. In comparison with the traditional optimization method, the time needed to finish the optimization procedure can be greatly reduced. The method has been validated by its application to the separation of several samples such as amino acid derivatives, aromatic amines, in which satisfactory separations were obtained with predicted resolution. PMID- 12541908 TI - [Transfer of solutes in capillary electrochromatography with mixed-mode stationary phase]. AB - In capillary electrochromatography with the mixed-mode stationary phase, while the transfer of solutes could be controlled by the contributions of chromatography (reversed-phase and ion exchange), the transfer of charged solutes could be influenced by electrophoresis transfer. According to the principle of ion independent transfer and several kinds of interaction between solute, mobile phase, and stationary phase, an theoretical expression was derived to describe the relationship among the apparent transfer velocity of solute, the transfer velocities in various forms and the diversified interactions. The studies focused on the influence of various parameters (e.g., pH, organic modifier content, stationary phase proportion) and the transfer characters of various solutes in the mixed-mode capillary electroosmotic chromatography (CEC). These results demonstrate that electroosmotic flow in the mixed stationary phase electrochromatography are kept high and steady within wide ranges of pH and organic modifier content. The separation was affected by the pH through the change of the solutes conformation. The influence of neutral solutes by organic modifier complies with the rule of conventional reversed-phase CEC. The separation of charged solutes was influenced evidently by the compete reagents which could improve the shape of the peak, but it was not so important as in the conventional ion exchange chromatography due to the adjustment by the function of electrophoresis. PMID- 12541910 TI - [Resolution of the epinephrine enantiomers derivatized with fluorescent chiral reagent by reversed-phase high performance liquid chromatography]. AB - A new method was developed to separate epinephrine enantiomers derivatized with fluorescent chiral reagent, R-(-)/S-(+)-4-(N, N-dimethylaminosulfonyl)-7-(3-iso thiocyanatopyrrolidino)-2, 1, 3-benzoxadiazole (DBD-PyNCS), by reversed-phase high performance liquid chromatography, on a Diamonsil C18 column(150 mm x 4.6 mm i.d., 5 microns), with a mobile phase of water-acetonitrile(72:28, volume ratio) and a fluorescent detector at the excitation wavelength of 460 nm and the emission wavelength of 550 nm. The column was eluted at 40 degrees C, with a flow rate of 1 mL/min. At first, let the fluorescent chiral reagent(36 mmol/L, 10 microL) react with epinephrine enantiomers(1 mmol/L, 10 microL), and then 10 microL 20% pyridine acetonitrile solution was added. The resulting mixture was stirred for 1 min and kept at 65 degrees C for 35 min and then a 10 microL aqueous of the diastereomeric derivatives was injected directly into the chromatograph. The diastereomeric compounds were efficiently separated. The Rs value was 2.6 with alpha of 1.07. The k's of L-epinephrine and D-epinephrine were 17.11 and 18.13 respectively. The retention times of L-epinephrine and D epinephrine were 30.24 min and 31.94 min respectively. The linear range was 0.07 g/L -0.50 g/L (r = 0.9995). The RSD was 4.3%(n = 7). The detection limit was 12 g/L(S/N = 3). PMID- 12541909 TI - [Application of fingerprint chromatogram in quality control of Shen-Mai injection]. AB - The theory and practice of traditional Chinese medicine require some comprehensive methods to assess quality of the Chinese herbal medication. Fingerprint chromatogram is one of the feasible approaches to evaluate the quality of Chinese herbal medication. So the fingerprint chromatogram of Shen-Mai injection was established by using reversed-phase high performance liquid chromatography. The chromatographic conditions were as follows: a Hypersil C18 column was used; the mobile phase was composed of water (A) and acetontrile (B) with linear gradient elution (0-50 min, 5%-95% B, volume fraction); the flow rate was 1.0 mL/min and the UV absorbance detection was set at 202 nm. The peak-area ratios of twenty-three fingerprint peaks and internal standard (diphenyl) were taken as the criteria for quality control. The quality differences in various batches and various manufacturers of Shen-Mai injections were investigated by projection discriminance based on principal component analysis. The results show the method developed is convenient, reliable and applicable for the quality control analysis of Shen-Mai injection. PMID- 12541911 TI - [Separation and purification of cellulase using affinity membrane]. AB - The importance of cellulase as a means for the efficient utilization of abundant cellulose resources in the world has been well recognized. Many researchers devote themselves to studying the mechanism of the action of cellulase to cellulose so that such expensive enzyme can be used much more widely. The first step is to obtain cellulase of high purity. So purification of cellulase is the key point in this field. However, the major problem in isolation is that cellulase is a complicated enzyme system and needs too many steps for separation, and that every cellulase needs special purification processing which cannot be used for the others. A novel method for the separation of the cellulase from crude extraction of Aspergillus niger with normal qualitative filter paper processed by 5 mol/L sodium hydroxide without precipitation and desalting steps was developed. Further purification of the cellulase was achieved by using an anion-exchange column of POROS 20HQ. The cellulase purified was identified as a new endoglucanase that had relatively high endurance to pH and temperature. Its relative molecular mass was estimated to be 60,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis. This enzyme exhibited very high activity towards carboxymethyl cellulose (CMC) with specific activity of 350 U.mg-1 and the recovery of activity of 9.7%. Its optimum pH and temperature were 4.0 and 70 degrees C, respectively. This is a simple, rapid and efficient method for purifying cellulase with high activity. PMID- 12541912 TI - [Alkene bromination used for detailed hydrocarbon and bulk hydrocarbon group-type analysis of gasolines containing alkenes]. AB - The optimized reaction conditions of selective alkene bromination for gasolines containing aromatics and saturated hydrocarbons are presented. By this way, the interfering problem in alkene determination from coeluting saturated hydrocarbons has been solved. So the detailed hydrocarbon analysis can be improved by a simple system containing polar and non-polar columns or by a gas chromatograph coupled with an atomic emission detector (GC-AED). Under the optimized conditions, it was found that the alkene compounds were selectively and completely brominated but the aromatics and alkane compounds were remained unaffected. A simple treatment, 90 s-120 s for reaction and 10 s-20 s for removing the excess bromine, can be easily realized. The treatment is applied for the different types of gasoline containing 0-100% alkene. Besides, one of the most important applications of this treatment is to analyse the hydrocarbons in detail from the fluid catalytic cracking (FCC) gasoline. The samples in these cases may not be accurately analyzed when using the traditional method of hydrocarbon analysis because of the presence of coeluted interfering olefins above C7. PMID- 12541913 TI - Method development of enantiomer separations by affinity capillary electrophoresis, cyclodextrin electrokinetic chromatography and capillary electrophoresis-mass spectrometry. AB - Capillary electrophoresis (CE) has become a powerful tool for enantiomer separations during the last decade. Since 1993, the author has investigated enantiomer separations by affinity capillary electrophoresis (affinity CE) with some proteins and by cyclodextrin electrokinetic chromatography (CDEKC) with some charged cyclodextrins (CDs). Many successful enantiomer separations are demonstrated from our study in this review article. In the enantiomer separations by affinity CE, the deterioration of detection sensitivity was observed under high concentration of the protein in running solutions. The partial filling technique was practically useful to solve the serious problem. It allowed operation at high protein concentrations, such as 500 mumol/L, without the detection problem. Charged CDs had several advantages for the enantiomer separations over neutral ones. Strong electrostatic interactions between a charged CD and oppositely charged analytes should be effective for the formation of the complex. A large difference in electrophoretic mobility between the free analyte and the inclusion complex should also enhance the enantiomeric resolution. In CE-mass spectrometry (CE-MS), the partial filling technique was applied to avoid the introduction of nonvolatile chiral selectors into the CE-MS interface. By replacing the nonvolatile electrolytes in the running buffer by volatile ones, the separation conditions employed in CE with the UV detection method could be transferred to CE-MS. PMID- 12541914 TI - [Characterization of chromatographic behavior of polar solutes of small molecules with lgI in reversed-phase liquid chromatography]. AB - A new characterization parameter lgI, affinity constant, in reversed-phase liquid chromatography (RPLC) has been tested at several aspects with experimental data of polar n-alkyl alcohol homologoues and aromatic alcohol homologoues. The lgI values were compared with other physicochemical parameters of corresponding solutes, such as hydrophobic fragmental constant (lgf), van der Waals volume (Vw), molecular connectivity (chi). It was proved that lgI was also used for the characterization of solutes, mobile and stationary phases in RPLC. Moreover, it was also confirmed that lgI has the property of thermodynamic equilibriam constant. PMID- 12541915 TI - [Design and evaluation of a confocal laser-induced fluorescence detector]. AB - A portable laser-induced fluorescence detector, based on confocal configuration detection system has been developed. This is assembled from commercially available components. All the components of the detector are domestic, which makes it low cost. The routine alignment procedure is simplified by using a skillful and visual alignment system and requires minimal experience for operation. The module design makes it possible for high performance liquid chromatographic, capillary electrophoretic and microfluid chip applications. The performance of the detector, including the sensitivity, noise, linear range and detection limit, was evaluated by capillary electrophoresis and flow injection analytical technique using a red-absorbing cyanine derivative (Cy5) and Cy5 labeled tryptophan as test samples. The results show that the background signal is very low and the peak-to-peak noise level is 0.002 mV. The detection limit and the linear dynamic range are 3.7 nmol/L and 10(3), respectively. PMID- 12541916 TI - [Determination of sildenafil in medicines for erectile dysfunction by capillary electrophoresis]. AB - A method has been developed for the determination of sildenafil in the medicines for erectile dysfunction by capillary electrophoresis. The samples were analyzed with 60 mmol/L NaH2PO4(pH 5.0) running buffer at 35 degrees C capillary temperature and 30 kV voltage. A linear calibration was obtained from 0.07 g/L to 1.05 g/L of sildenafil (r = 0.9985) with the RSD of 4.7% for peak area, and the average recovery was 97.4%. The results were compared with those of HPLC, showing that the method is precise, simple and cost-effective, and can be used as a complementary method to HPLC. PMID- 12541917 TI - [Chiral separation of enantiomers of synthesized amlodipine and its intermediate by capillary electrophoresis]. AB - A capillary electrophoretic method for the chiral separation of enantiomers of synthesized amlodipine and its intermediate was developed. Several cyclodextrins (CDs) were applied as the chiral selectors and it was found that the ionic modified carboxymethyl-beta-cyclodextrin (CM-beta-CD) could give satisfactory enantioselectivity. In addition, the effects of the pH value of the buffer system, the concentration of the CD and the voltage on the chiral separation were investigated. The optimized buffer for amlodipine and its intermediate enantiomers was a buffer containing 30 mmol/L phosphate and 50 mmol/L CM-beta-CD (pH 6.12). Under these conditions, the resolutions of enantiomers of amlodipine and its intermediate were 1.73 and 1.55, respectively. PMID- 12541918 TI - [Analysis of carbaryl and other pesticides by capillary electrophoresis]. AB - Five pesticides, i.e. carbaryl, simazine, atrazine, arnetryne and prometryn were separated by capillary electrophoresis technique. The effects of experimental parameters including pH, the concentration of surfactant and organic modifier on the separation were investigated. From the experiments, we found the optimum electrophoretic conditions were as follows: 20 mmol/L borate-50 mmol/L sodium dodecyl sulfate-10% methanol (volume fraction) as the running buffer (pH 9.2), applied voltage of 25 kV, detection wavelength of 225 nm. Under the optimum conditions, baseline-separation of each component has been obtained using anionic surfactant for micellar electrokinetic chromatography. The results indicated that the calibration curve of carbaryl was linear in the range of concentrations 30 mg/L-200 mg/L. This method has been applied to the determination of real sample with satisfactory results obtained, and the recovery of carbaryl was 93.4% 101.3%, and the relative standard deviation of peak area was 2.59%. PMID- 12541919 TI - [Analysis of a novel platinum anticancer compound HJ5 by high performance liquid chromatography]. AB - Cis-dichloroamine (4-amino-2, 2, 6, 6-tetramethylpiperidine-1-oxyl) platinum (II) (abbreviated to HJ5) is a novel platinum anticancer compound which has prospects for research and application. In order to establish a conventional analytical method, an HPLC has been studied. Under the conditions of acetonitrile-water (10:90, volume ratio) as mobile phase, detection wavelength of 210 nm, a Phenomenex column C18(150 mm x 3.9 mm i.d., 10 microns). The linear equation, recovery and accuracy of the method were determined with external standard method. The HPLC has satisfactory resolution between the peaks of HJ5 and impurities. The peak areas of HJ5 were linear to its amounts detected and the detection limit was 0.07 microgram. The hydration of HJ5 can be inhibited effectively by NaCl solution. In a 9 g/L NaCl solution, the stability can be kept for more than 2 hours which is enough for analysis. PMID- 12541920 TI - [Determination of the base contents of liver DNA of rats by reversed-phase high performance liquid chromatography]. AB - The base contents of liver deoxyribonucleic acid (DNA) of rats living at an altitude of 2.3 km were determined by reversed-phase high performance liquid chromatography. At first, 0.05 mol/L KH2PO4(pH 4.0) was used to dissolve the DNA acid hydrolysis products with 8-bromoguanosine (Br8G) as an internal standard. Then the DNA hydrolysis products with Br8G were chromatographed on a Supelcosil LC-18 column with UV detection at 254 nm and eluted by the mobile phase of MeOH 0.05 mol/L KH2PO4(pH 4.0) (20:80, V/V) at the flow rate of 0.8 mL/min. Under these conditions, several bases were separated effectively. From the results, the relatively constant proportions of the bases in DNA were found. The contents were 17.4% of cytosine (C), 28.8% of adenine (A), 23.3% of guanine (G) and 25.3% of thymine (T). RSDs of the determination of these bases were 1.7%, 1.5%, 1.3% and 2.1%, respectively. At the same time the methylation level of liver DNA of the rats determined by the internal standard method was 6.2%. PMID- 12541921 TI - [Determination of furazolidone, carbenoxolone sodium and berberine hydrochloride in wei kang tablets by reversed-phase high performance liquid chromatography(RP HPLC)]. AB - An HPLC method with gradient elution was developed to separate and determine furazolidone, carbenoxolone sodium and berberine hydrochloride in Wei Kang Tablets on Nova-Pak C18 column (150 mm x 3.9 mm i.d., 4 microns), using acetonitrile-phosphate buffer solution (pH 7.0) as mobile phase and 3,5 dinitrobenzoic acid as the internal standard. Detection was performed with UV detector at 254 nm. The calibration curves were linear within the ranges of 141.2 mg/L -1,270.8 mg/L for furazolidone(r = 0.9997), 100.6 mg/L -905.4 mg/L for carbenoxolone sodium(r = 0.9995) and 99.2 mg/L -892.8 mg/L for berberine hydrochloride(r = 0.9991). The recoveries and RSDs were 99.5% and 1.38% for furazolidone, 100.3% and 1.73% for carbenoxolone sodium, 97.3% and 1.97% for berberine hydrochloride respectively. The results show that this method is simple, rapid, specific, accurate and reproducible. PMID- 12541922 TI - [Determination of purity of different types of aprotinin by micellar electrokinetic capillary chromatography]. AB - A sensitive and rapid method was developed for purity determination of different types of aprotinin by micellar electrokinetic capillary chromatography (MECC). MECC was performed with 80 mmol/L Na2HPO4-H3PO4(pH 7.00) buffer solution containing 4 mmol/L cetyltrimethyl ammonium bromide (CTAB) at an applied voltage of 15 kV. The electrophoresis was monitored at 277 nm, and completed in 6 minutes. The relative standard deviations of migration time and area for aprotinin were below 0.3% and below 5% respectively within 1 day (for 6 repetitive runs). The effect of high concentration of salt in the samples has been studied. The results obtained by capillary zone electrophoresis, MECC and high performance liquid chromatography were compared, and those of MECC were the best. PMID- 12541923 TI - [Simultaneous analysis of aspartic acid and sorbitol in potassium magnesium aspartate and sorbitol injection by ion-exclusion chromatography with evaporative light-scattering detection]. AB - An ion-exclusion chromatographic (IEC) method with evaporative light-scattering detector (ELSD) was established for the simultaneous analysis of the aspartic acid and sorbitol in Potassium Magnesium Aspartate and Sorbitol Injection. This procedure eliminates the need for complex derivatization schemes since ELSD can detect aspartic acid and sorbitol directly. An ion exclusion column (TSKgel OApak A column, 7.8 mm i.d. x 300 mm, 5 microns, Tosho, Japan) was used for separation. The mobile phase was a mixture of aqueous solution (containing 0.1% (volume fraction) trifluoroacetic acid)-methanol (9:1, volume ratio). Under the above conditions aspartic acid and sorbitol were separated to baseline in 12 min. Regression equations revealed linear relationship (correlation coefficient: 0.999) between the concentrations of the analytes injected and the peak areas of the analytes detected by ELSD. The detection limits of ELSD (S/N = 3) were about 30 mg/L and 20 mg/L for aspartic acid and sorbitol, respectively. PMID- 12541924 TI - [Development of gas chromatographic method for chiral analysis of acrylic ketone alcohol and acrylic ketone acetate]. AB - A gas chromatographic method was developed to directly determine the enantiomers of acrylic ketone alcohol and acrylic ketone acetate. Two pairs of enantiomers were nearly baseline-separated by using Model HP-5890 Gas Chromatograph with flame ionization detector and WCOT CD-Chiral-DEX CB (30 m x 0.25 mm i.d.) column. The optimum conditions were the split ratio of 30:1 for the sample injected, the flow rates of 3.7 mL/min for the carrier gas (nitrogen), 60 mL/min for the hydrogen, and 250 mL/min for the oxygen, and temperatures of 148 degrees C for the column, 230 degrees C for the injector, and 230 degrees C for the detector. The linearity and reproducibility were satisfactory. The regression coefficients were over 0.9965. The RSDs obtained were less than 1.25%. The method was further used to trace the reactants and products, and to evaluate the enzyme catalyst activity and selectivity under different conditions. The results show that the method developed is very successful. PMID- 12541925 TI - [Gas chromatographic determination of micro organic impurities in 3-chloro-2 hydroxypropyl trimethyl ammonium chloride]. AB - A simple, rapid and reliable method for the determination of micro organic impurities, epichlorohydrin and 1,3-dichloropropanol, in 3-chloro-2-hydroxypropyl trimethyl ammonium chloride (CHPTMA) by gas chromatography is reported. 2 Ethylhexanol was the internal standard. Epichlorohydrin and 1,3-dichloropropanol in CHPTMA were extracted by chloroform. The gas chromatographic separation was achieved on a stainless steel column packed with 10% PEG 20M/Chromosorb W/AW and with a flame ionization detector. The ranges of recovery were 97.5%-105.0% and 93.3%-98.8%, the RSDs were 11.5% and 13.1% and the mininum detectable concentrations were 5.0 micrograms/g and 10 micrograms/g for epichlorohydrin and 1,3-dichloropropanol respectively. PMID- 12541926 TI - [Analysis of mannoligosaccharides by liquid chromatography-electrospray ionization mass spectrometry]. AB - Oligosaccharide characterization has been of utmost interest in various areas such as medicine, biochemistry, and food chemistry. These biologically relevant molecules are ideally suited for mass spectrometric investigation, because of the capability of this technique in offering structure and relative molecular mass information. Therefore, liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) was applied to characterize the acetolysis of mannan from Saccharomyces cerevisiae. The electrospray using Na+ as adducts proved to be superior to the LC-MS for the determination of mannoligosaccharides. LC separation was accomplished by the use of NH2 column and the elution by acetonitrile-water (70:30, volume ratio). The results showed that mannoligosaccharides side chain consisted of mannose, mannobiose, mannotriose and mannotetraose. The method developed is accurate, fast and convenient and can be used to characterize the relative molecular mass of the oligosaccharides. PMID- 12541927 TI - [Quantitative determination of barbaloin in aloe capsule by high performance liquid chromatography]. AB - Barbaloin in aloe capsule was quantitatively determined by high performance liquid chromatography with ODS column, a mixture of CH3OH-H2O (50:50, volume ratio) as mobile phase and UV detection at 298 nm. There was a good linear relationship within the range of 0.02 microgram-1.50 micrograms (r = 0.9999). The recovery was 99.0%-101.9% and the relative standard deviation was 1.37%(n = 6). The detection limit was 0.002 microgram (S/N = 2:1). The total time for separation and determination was within 10 min. The method is rapid, simple, accurate and reproducible. The method is proposed to be used for quality control of aloe capsule. PMID- 12541928 TI - [Simultaneous determination of 18 amino acids by reversed-phase high performance liquid chromatography with precolumn phenylisothiocyanate derivatization]. AB - A reversed-phase high performance liquid chromatographic method for simultaneous determination of 18 amino acids is described. The sample was mixed with norleucine as the internal standard, then derivatized with phenylisothiocyanate (PITC) and analyzed on a Kromasil C18 column at 38 degrees C, using gradient elution with detection at 254 nm. The correlation coefficients between the ratios of peak area of amino acid to that of the internal standard and the amino acid concentrations were above 0.99, except for cysteine(Cys)(0.962). The recoveries of amino acids added were 96.0%-102.4%. Detection limit of leucine was 0.5 mg/L. The method was applied to analyze free amino acids in deproteinized calf blood injection and good results were obtained. PMID- 12541929 TI - [Determination and stability tests of triclosan in disinfectants]. AB - A method has been developed for the determination of triclosan in antibacterial detergent by RP-HPLC equipped with a diode-array detector. A Kromasil C18 column (200 mm x 4.6 mm i.d., 7 microns) was used with a mobile phase of methanol acetonitrile-water (40:40:20, volume ratio) containing 0.02 mol/L KH2PO4 at pH 2.7 and UV detection at 280 nm. The samples were dissolved with the mobile phase by ultrasonic method. The recoveries of triclosan were from 94.2% to 102.2% and the RSDs were from 2.2% to 3.0%. The method is simple, rapid, accurate and reproducible. The concentrations of standards and samples did not change evidently after storage at 54 degrees C for 14 d and at 37 degrees C for 90 d. The results showed that triclosan has a good stability. PMID- 12541930 TI - [Determination of 2,6-di-tertbutyl-4-methylphenol in an aviation jet fuel by high performance liquid chromatography]. AB - The content of antioxidant of 2,6-di-tertbutyl-4-methylphenol(BHT) in a new aviation jet fuel (HDF-1) was determined by high performance liquid chromatography. The influences of the volume ratio of methanol and acetate buffer and the flow rate for BHT separation from HDF-1 were studied and the best analysis conditions were obtained. The optimum volume ratio is 85:15 and the optimum flow rate is 1 mL/min. The relation of BHT mass fraction and its peak area has been analyzed and simulated. It was found that they had a good linear relationship. PMID- 12541931 TI - [Determination of volatile constituents in guanxin suhe wan by gas chromatography mass spectrometry]. AB - The volatile constituents of Guanxin Suhe Wan and its ingredient drugs were analyzed by gas chromatography-mass spectrometry. Two compounds, borneol and benzyl benzoate were determined by selected ion monitoring with methyl salicylate as the internal standard. The recoveries of borneol and benzyl benzoate were 91.7% and 89.7% with the RSDs of 5.6% and 2.3%, respectively. PMID- 12541932 TI - [Simultaneous determination of fluoroacetamide and tetramine by gas chromatography]. AB - A simple and fast gas chromatographic method was used for the simultaneous determination of fluoroacetamide and tetramine. The samples were extracted with acetone and the extracts were analyzed by gas chromatography. A megabore capillary column coated with FFAP and a nitrogen-phosphorous detector were used to obtain satisfactory selectivity and sensitivity. The detection limits and average recoveries were 5 ng and 69.1% for fluoroacetamide and 1 ng and 89.8% for tetramine. PMID- 12541933 TI - [Determination of diniconazole residue in grape by gas chromatography]. AB - An efficient method was developed for the determination of diniconazole in grape by gas chromatography. The diniconazole residue in the sample was extracted by acetone. The extract was cleaned-up by liquid-liquid extraction and then solid phase extraction. The diniconazole residue was separated completely from other impurities in the sample through SPB -1701 capillary column (30 m x 0.53 mm i.d. x 1.0 micron) and determined by an electron capture detector. The recoveries ranged from 85.8% to 94.7% when the added levels were from 0.01 mg/kg to 1.00 mg/kg. The RSD was within 5% and the limit of detection was 0.01 mg/kg. PMID- 12541934 TI - [Studies on the retention mechanism of N-methylaniline bonded silica for high performance liquid chromatography]. AB - Three N-methylaniline bonded stationary phases containing different bonding amounts were prepared by modifying silica gel with N methylanilinetriethoxysilane. Influences of the bonding amounts on the retentions and selectivity of polycyclic aromatic hydrocarbons(PAHs), polar-, acid- and basic-substituted aromatic compounds were investigated in reversed- and normal phase high performance liquid chromatography. In the reversed-phase liquid chromatography, the plots of logk' against the carbon atom numbers for alkylbenzenes are linear, and the retentions and relative retentions of the solutes(including alkylbenzenes, PAHs and nitrobenzenes) increased with the increase of the bonding amounts and water volume fractions in mobile phase. The results indicated that the hydrophobic interaction played a major role in the retention process. In the normal-phase liquid chromatography, the retentions of PAHs increased and the retentions of nitrobenzenes decreased with the increase of bonding amounts. With the increase of bonding amounts, the retentions of acidic compounds increased and the retentions of basic solutes decreased considerably. It is indicated that the number of imino groups increased and the activity of residual silanols diminished with the increase of bonding amounts. The retentions of solutes on the stationary phases result from acid-basic, dipole-dipole, pi-pi and charge transfer interactions in the normal phase chromatography. PMID- 12541935 TI - [Study on the production of trehalose by bacterium D-97 endocellular enzymes using HPLC/RI and HPLC/ESI-MS]. AB - The mechanism in which trehalose is produced from dextrin or starch hydrolyzate by endocellular enzymes of bacterium D-97 can be elucidated high performance liquid chromatography (HPLC) with differential refraction detection (RI) basically, including the effect of the different carbon sources on the endocellular trehalose-producing enzymes in bacterium D-97 and the possibility or ability of the endocellular enzymes to produce trehalose using maltooligosaccharides of different chain lengths. After purification of endocellular enzymes of bacterium D-97, two enzymes (called Enzyme A and Enzyme B) related to trehalose synthesis were found. The unknown oligosaccharides produced by Enzyme A were analyzed with the HPLC/RI and HPLC/ESI-MS (electrospray ionization mass spectrometry). The results showed the relative molecular masses of the unknown oligosaccharides were the same as those of the enzymatic reaction substances (maltotriose, maltotetraose and maltopentaose) respectively. In combining with other results of biological experiments, these unknown oligosaccharides had been identified basically. There was no reduction power in these unknown oligosaccharides and only one trehalose residue exited in the molecular chain of these unknown oligosaccharides. PMID- 12541936 TI - Preparation and characteristics of new coating for solid-phase microextraction. AB - Poly (methylphenylvinylsiloxane) (PMPVS) coating for solid-phase microextraction (SPME) was first prepared by using sol-gel approach and cross-linking of free radical initiation. The extraction properties of the novel coating for aromatic compounds were investigated using a self-made SPME device coupled with a gas chromatograph-flame ionization detector (GC-FID). The coating provided high surface areas and allowed high extraction efficiency. Compared with some commercial SPME stationary phases, the new phase showed better selectivity and sensitivity toward aromatic compounds. Furthermore, PMPVS coating showed good thermal stability and longer lifetime. PMID- 12541937 TI - Gas chromatographic analysis of aromatic hydrocarbons in gasoline by column switching-back flushing technique and normalization method. AB - A simple and efficient method based on the technique of capillary column switching-back flushing was developed for the detailed analysis of aromatic hydrocarbons in gasoline. The early eluting components from a pre-column and the components of interest eluting from an analytical column are all directed to a flame ionization detector through a tee piece, which facilitates the quantitation of aromatic hydrocarbons in gasoline by normalization method. Real samples were analyzed and RSD < or = 3%. The operation is simple and easy, and the switching time window is +/- 5 s. PMID- 12541938 TI - [Solid-phase microextraction for the analysis of drugs and biological samples]. AB - Solid-phase microextraction (SPME) as a new sample pretreatment technique was developed in 1990s. The applications of SPME on drugs and biological samples are reviewed with 60 references. The references were classified into twelve groups according to the character of analysis. PMID- 12541939 TI - [Application of resilient backpropagation neural network in predicting hydrophobic parameters of alkylbenzenes]. AB - Artificial neural networks have been applied for predicting the hydrophobic parameters of alkylbenzene. Compared with traditional methods it has the advantages of simple operation and wide applications. Based on error back propagation neural networks the relationship among the molecular connectivity index (chi), van der Waals surface area (Aw) and hydrophobic parameter was studied, meanwhile the mathematical model was established and used to predict the hydrophobic parameters. By comparing the hydrophobic parameters of experimental values with those calculated by neural networks, we found they had good agreement. The average relative deviation was less than 1%. Because traditional back propagation network is generally time consuming, resilient backpropagation (RPROP) algorithm was used to solve this problem. By using RPROP algorithm, the hydrophobic parameters were obtained precisely by fast training and simple parameter's selection. It needed less than 1,000 iterations to reach the goal on the computer operated at 1.4 GHz. The present work shows that the artificial neural network is a new powerful tool to predict the physicochemical parameters. PMID- 12541940 TI - [Simple preparation and evaluations of amide-octyl-bonded phase]. AB - A simple preparation process of amide-octyl-bonded phase (AOBP) is described. In the process aminopropyltriethoxysilane first reacted with octanoyl chloride and then the obtained intermediate was bonded onto porous silica. Characterization of prepared packing was carried out with elemental analysis, solid-state nuclear magnetic resonance (13C-NMR) and Fourier transform infrared (FTIR) spectroscopy. Chromatographic evaluations were carried out by testing with a mixture including acidic, basic and neutral organic compounds using methanol-water as binary mobile phase. The results show that the stationary phase has excellent chromatographic properties and resistance to hydrolysis between pH 2.5-7.5. It can be used for the efficient separation of basic compounds. PMID- 12541941 TI - [Effect of the bonding amounts of chiral ligand exchange chromatographic stationary phases on the enantioseparation of amino acids]. AB - L-proline chiral stationary phases with various bonding amounts for ligand exchange chromatography were prepared by treating silica gel with 3 glycidoxypropyltrimethoxysilane and bonding L-proline. The bonding amounts of L proline chiral stationary phase were controlled effectively and characterized by IR and elemental analysis. Some DL-amino acids were separated and the influences of the bonding amounts on the efficiency of resolution of DL-amino acids by ligand exchange chromatography were briefly discussed and explained by the mechanism of the separation. PMID- 12541942 TI - [Development of capillary electrophoretic method for determination of the inorganic anions in the ash of black powder]. AB - A capillary electrophoretic method for determining the inorganic anions in the ash of black powder has been developed. The components and pH of the buffer solution, the concentration of osmotic flow modifier and the separation voltage were investigated. The optimized parameters were 5.0 mmol/L sodium chromate buffer solution (pH 8.20) containing 0.5 mmol/L cetyltrimethylammonium bromide (CTAB) as osmotic flow modifier, a separation voltage of -20 kV and a detection wavelength of 254 nm. Under the specified conditions five anions were completely separated in four minutes, and the repeatabilities (RSD) of migration time and peak area were in the range of 0.17%-1.4% and 3.9%-5.0%, respectively. The detection limits were in the range of 5.0 mumol/L to 10.0 mumol/L. To show the usefulness, the method was applied to analyze the ash of a black powder and the results showed that the relative standard deviations of the determination for Cl- and NO2- were 6.0% and 3.9%, respectively. PMID- 12541943 TI - [Separation of saccharides in tobacco by capillary zone electrophoresis]. AB - A method for the separation of derivatized saccharides in tobacco with high performance capillary electrophoresis and ultraviolet spectrophotometry (HPCE-UV) is presented. Five saccharides in tobacco were derivatized with p aminobenzonitrile, separated at pH 10.5 with 50 mmol/L borate buffer containing 5% (volume fraction, the same in the following percentage concentration) methanol, 5% acetonitrile, 2.5% ethyleneglycol, 2.5% 2-propanol and 1 mmol/L sodium dodecyl sulfate, and detected at 285 nm. PMID- 12541944 TI - [Research on the separation of caffeine and its nine analogues by micellar electrokinetic capillary chromatography]. AB - The separation of caffeine and its nine analogues by micellar electrokinetic capillary chromatography (MECC) using sodium dodecyl sulphate (SDS) as the micellar phase was investigated. The effects of pH and concentration of phosphate buffer solution, SDS micelle concentration, methanol volume fraction, applied voltage and temperature on the separation were studied. It was found that the migration of these compounds was affected by these factors, especially by pH of the solution. The order of elution, as well as the migration time and separation efficiency of these compounds changes with the acidity of the solution, which suggests the disassociation of the hydrogen on the secondary amino group in these compounds. The optimized separation conditions consisted of a running buffer of 20 mmol/L sodium phosphate buffer, at pH 11.0, containing 20 mmol/L SDS, with an applied voltage of 25 kV and a temperature of 25 degrees C. All of the compounds were resolved within 9 min with the detection limits of 0.70 mg/L and the linearity in the range of 1.40 mg/L-45.5 mg/L with the correlation coefficients of 0.998-0.999. PMID- 12541945 TI - [Investigation of separation mechanism for neutral solutes on cyano column in capillary electrochromatography]. AB - The separation mechanism for neutral solutes on a cyano column in capillary electrochromatography (CEC) was investigated and the effects of components of mobile phases and the kinds of buffers on retention of samples were studied. In comparing the separation behavior of samples on CN column with reversed-phase ODS column and normal phase SI column, the peak order was different on CN column under different experimental conditions, and the characteristics of CN column having both reversed and normal phase mechanisms were obvious. The migration velocities for neutral solutes of different polarity on CN column were very different by the effects of two mechanisms under different operating conditions. It is easy to change peak order and to adjust selectivity. PMID- 12541946 TI - [Influence of mobile phase composition on chiral separation of organic selenium racemates]. AB - The chiral separation of some chiral compounds with similar structure on the cellulose tris (3,5-dimethylphenylcarbamate) chiral stationary phase prepared by us was obtained. Ternary mobile phases influencing chiral recognition were investigated. A mode of interaction between the structural character of samples and chiral stationary phase is discussed. The results indicated that the retention and chiral separation of the analytes had a bigger change with minute addition of alcohols or acetonitrile as modifier in n-hexane/2-propanol (80/20, volume ratio) binary mobile phase. PMID- 12541947 TI - [Studies on the separation and determination of natural phenolic acids by reversed-phase high performance liquid chromatography]. AB - A new method using gradient elution of reversed-phase high performance liquid chromatography was set up for the separation and determination of 12 natural phenolic acids simultaneously. The separation conditions were studied with a Zorbax ODS (5 microns) column, and optimized conditions were suggested with the mobile phase gradient elution of CH3CN-H2O (with 0.5% HAc) (5:95-->35:65, V/V, in 40 min) at room temperature, flow rate at 1.0 mL/min and detection wavelength at 280 nm. And the effects of acidity on separations and the relationships between the phenolic acids structures and their retention behaviors were discussed. Qualitative and quantitative analysis of the 12 compounds were performed. The regression equation, correlation coefficient, linear range, detection limit and relative standard deviation (RSD) of each compound were obtained under the optimized conditions. The correlation coefficients of the 12 compounds were between 0.9980 and 0.9999. The detection limits ranged from 0.96 ng to 4.10 ng and RSDs were < or = 2.64%. These results demonstrate that the optimized determination method obtained is successful for the separation and determination of the 12 phenolic acids analysed. PMID- 12541948 TI - [Separation of different phosphatidyl cholines by reversed-phase ion-pair high performance liquid chromatography]. AB - A reversed-phase ion-pair high performance liquid chromatographic (RP-IP-HPLC) method for the separation of different phosphatidyl cholines (PC) was established. The optimized conditions were a PERKIN-ELMER/HS-5 C18 column (150 mm x 4.6 mm i.d., 5 microns), isocratic elution with mobile phase of methanol acetonitrile-water (70:22:8, volume ratio) containing 15 mmol/L tetramethyl ammonium phosphate (TMAP) at pH 7.0, flow rate of 2.0 mL/min and UV detection at 208 nm. Seven kinds of PC could be separated with this method. PMID- 12541949 TI - [Determination of aconitine alkaloids in a pesticide--0.25% aconitine alkaloids mixed with emulsion by high performance liquid chromatography]. AB - The raw root of Aconitum, an important Chinese traditional medicine, contains some very toxic alkaloids such as aconitine, mesaconitine and hypaconitine etc. They are usually processed to lower the alkaloid content before used as a drug. The extract of crude drug (aconite) can be made into a plant pesticide by means of mixing with some emulsion. In order to evaluate the quality of the pesticide, we developed a rapid, specific and precise method using high performance liquid chromatography (HPLC) for the separation and quantitation of the alkaloids in the aconite extract and the mixed products (the aconite extract is mixed with emulsion). Before the determination by HPLC, the sample must be acidified with 2% (mass percentage) HCl at first. Extract the acid liquid with CHCl3. Alkalize the extract with ammonia, and add a little of 0.1 mol/L NaHCO3 and 0.1 mol/L Na2CO3 till pH 9. Extract it with CHCl3. Evaporate the extract and add a certain amount of methanol. Add the internal standard into the sample. Inject the combined sample solution onto a column of chemically bonded octadecylsilane phase and develop the chromatogram with MeOH-H2O-CHCl3-triethylamine (68:32:2:0.1, volume ratio). Aconitine, mesaconitine, hypaconitine and medroxyprogesteroni acetas (internal standard) were on base line separated. Quantify the alkaloids by peak area ratio (aconitine alkaloids vs internal standard). This method has high recovery (> 92%) and good reproducibility (RSD < 3.2%). PMID- 12541950 TI - [Determination of zopiclone in serum by reversed-phase high performance liquid chromatography]. AB - A reversed-phase high performance liquid chromatographic method (external standard method) was developed for the determination of zopiclone in serum. After the selective extraction with n-butyl chloride, this compound was chromatographed on a LiChroCART 125-4 column packed with LiChrospher 60 RP select B(5 microns) using acetonitrile-monopotassium phosphate (20:80, V/V) as mobile phase. The eluting compound was measured by an ultraviolet detector at 254 nm. Spiked with zopiclone of 400 micrograms/L, 1,600 micrograms/L and 6,400 micrograms/L in serum, the recoveries of zopiclone were (73.4 +/- 3.2)%, (82.2 +/- 4.1)% and (90.3 +/- 4.5)% respectively. The detection limit of the method was 15 micrograms/L. The method is simple and rapid for the determination of zopiclone in forensic toxicology. PMID- 12541951 TI - [Separation and purification of lysozyme from egg white by high performance cation-exchange chromatography]. AB - A new method used to separate and purify lysozyme from egg white by high performance cation-exchange chromatography has been established. The process conditions for purifying lysozyme were also discussed in detail. The procedure involved that homogenization of the egg white sample, preliminary purification with sodium chloride, and chromatographic separation by the weak cation exchange column (XIDACE-WCX). The experimental results showed that the purified lysozyme and other impurity proteins were completely separated. By using bioactivity assay, the recovery of lysozyme was 107%, and the specific activity was 15,467 U/mg through the column. Its purity was raised 5.6-fold. The collected fraction with activity was detected by size-exclusion chromatography (SEC). The purified lysozyme was homogeneous. Compared with the traditional soft-based low pressure ion-exchange chromatography, the developed method is rapid and effective. PMID- 12541952 TI - [Determination of chloride and trace inorganic anions in natural gas field water with high chloride concentration by ion chromatography]. AB - An ion chromatographic method for determining chloride and trace anions, such as F-, Br-, I- and SO4(2-) in natural gas field water with high chloride concentration was studied. The separation of the anions was achieved on an IonPac AS11 column with NaOH as gradient eluent, and the detection was performed by a conductivity detection mode (self-regenerated suppression). The pre-treatment of the samples was through a filter of 0.45 micron and a Dionex OnGuard RP C18 column. The method is simple and useful, and has good linearity with correlation coefficients between 0.9994 and 0.9999. The recoveries of anions in the samples were between 98.4% and 102.1%. PMID- 12541953 TI - [On-line determination of transitional metal ions by low-pressure ion chromatography with chemiluminescence detection]. AB - The separation and determination of five transitional metal ions, Co2+, Cu2+, Mn2+, Fe2+ and Cr3+ by low-pressure ion chromatography with chemiluminescence detection were investigated for the first time. The separation was carried out on a C3 ion chromatographic column by using a step gradient elution with the mobile phase of oxalic acid-citric acid solution. The five transitional metal ions could be separated simultaneously. Luminol-H2O2-Mn+ system was used as the flow injection chemiluminescence analytical system. Under the optimum conditions with 0.5 mmol.L-1 luminol, 0.05 mol.L-1 H2O2 and 0.020 mol.L-1 NaOH, the linear ranges (mg.L-1) for Co2+, Cu2+, Mn2+, Fe2+ and Cr3+ were 0.001-0.1, 0.1-6, 0.06-4, 0.03 5 and 0.025-1 respectively; the detection limit (microgram.L-1) for Co2+, Cu2+, Mn2+, Fe2+ and Cr3+ were 0.85, 85, 42, 21 and 20. The present method is simple, rapid, accurate and has been successfully applied to detect Co2+ in vitamin B12 and Co2+, Cu2+, Mn2+, Fe2+ and Cr3+ in human hair sample with satisfactory results, which were in good agreement with those from atomic absorption spectrometry. PMID- 12541954 TI - [Determination of acetylene and vinyl chloride in hydrochloric acid from producing vinyl chloride by headspace gas chromatography]. AB - A headspace gas chromatographic method was used for determining acetylene and vinyl chloride in hydrochloric acid from vinyl chloride production. Hydrogen chloride was neutralized by sodium hydroxide, so its effect on determining acetylene and vinyl chloride was eliminated. The headspace bottle was put in a water bath at 35 degrees C for 45 min. The GC column was a 2 m x 3 mm i.d. stainless steel tube packed with GDX-202 stationary phase. The column temperature was 140 degrees C. Injection volume was 1 mL. External standard method was used for quantitation. The relative standard deviation was 0.85% and the recoveries in the range of 30.0 micrograms/g-150 micrograms/g were 98.9%-103% for acetylene. The relative standard deviation was 1.4% and the recoveries in the range of 20.0 micrograms/g-100 micrograms/g were 98.8%-102% for vinyl chloride. PMID- 12541955 TI - [Determination of p-tert-butylcatechol in styrene monomer by gas chromatography/mass spectrometry]. AB - An efficient method for the determination of p-tert-butylcatechol (TBC) in styrene monomer by gas chromatography/mass spectrometry (GC/MS) has been established. The sample was injected directly, and separated by gas chromatography with capillary column (HP-1, 30 m x 0.32 mm i.d. x 0.25 micron), then determined by a mass-spectrometer with electron impact (EI) at selected ion monitoring (SIM) mode. There was a good linear relationship within the range of 5 mg/kg-50 mg/kg (r2 = 0.9987). Compared with the colorimetric determination by a spectrophotometer (ASTM D4590), the method was proved to be fast and accurate. PMID- 12541956 TI - [Analysis of preservatives in cosmetics by gas chromatography]. AB - The wide bore capillary column HP-1701(15 m x 0.53 mm i.d. x 1.0 micron) has been used to analyze benzyl alcohol, methyl-p-hydroxy benzoate, ethyl-p-hydroxy benzoate, propyl-p-hydroxy benzoate, and butyl-p-hydroxy benzoate in cosmetics. The results showed that these preservatives could be separated with the capillary column completely and their peaks were not interfered with those of other components in cosmetics. The detection limits were 13 ng, 70 ng, 70 ng, 100 ng and 100 ng respectively and the linear range was 50 ng-500 ng. PMID- 12541957 TI - [Determination of aspirin and free salicylic acid in lysinipirine injection by high performance liquid chromatography]. AB - The contents of aspirin and free salicylic acid in lysinipirine injection were determined by high performance liquid chromatography (HPLC). A Hypersil BDS C18 column was used with the mobile phase of methanol-water-acetic acid (35:65:3, volume ratio) and the detection wavelength of 280 nm. The average recoveries of aspirin and salicylic acid added were 99.27% (RSD = 0.8%) and 99.61%(RSD = 1.3%), respectively. The calibration curves had good linearity in the range of 0.028 g/L -0.141 mg/L and 0.77 mg/L -3.85 mg/L, and the correlation coefficients were 0.9999 and 0.9998 for aspirin and salicylic acid respectively. PMID- 12541958 TI - [Preparation of capillary gas chromatographic columns for separation of polar organic compounds by sol-gel method]. AB - A sol-gel method was used in preparing capillary gas chromatographic columns with simplicity and rapidity. Due to the formation of the chemical bonds between the coating layer and the inner wall of the capillary, the thermal stability of the column was greatly increased. The sol-gel poly(dimethyl-siloxane) (PDMS) column was able to withstand temperatures as high as 400 degrees C. This column showed excellent separation of free fatty acids, amines, and other polar compounds. The sol-gel column also showed good repeatability of retention time on a single column as well as good repeatability of capacity factor, column efficiency, symmetry and McReynold's constant on several columns of the same type. PMID- 12541959 TI - [Analysis of sulfur compounds in residue fluid catalytic cracking gasoline]. AB - Sulfur compounds in residue fluid catalytic cracking (RFCC) gasolines from Refinery of SINOPEC Beijing Yanshan Company and Refinery of Shijiazhuang were analyzed by gas chromatograph connected with flame photometric detector (FPD) and atomic emission detector (AED). Twelve and 26 kinds of sulfur compounds were detected by AED in RFCC gasolines from Yanshan and Shijiazhuang respectively. Only 0 and 19 kinds of sulfur compounds were found by FPD in these two gasolines respectively. The experimental results demonstrated that AED is more sensitive and selective to sulfur compounds than FPD. It also indicated that thiophenes were the major sulfur compounds in the RFCC gasoline. In addition, mercaptan, sulfoether and disulfide species were found. PMID- 12541960 TI - [Study on the chemical constituents of the volatile oil from Lysimachia trientaloides Hemsl]. AB - The volatile oil from Lysimachia trientaloides Hemsl. was obtained by steam distillation. The chemical constituents were separated and identified by gas chromatography-mass spectrometry (GC-MS). The relative contents in the volatile oil were determined by peak area normalization. The yield of oil by steam distillation was 0.11%, and 40 chemical constituents were separated and identified. Terpenic series and their oxo-derivatives are major chemical constituents in the oil. The main compounds were patchouli alcohol(22.54%), L bornyl acetate(16.17%), gamma-gurjunene(3.27%), delta-guaiene(2.62%), nerolidol(2.02%), linalool(1.99%) and palmitic acid(1.96%). PMID- 12541961 TI - [Application of genetic algorithm in optimization of mobile phase composition in high performance liquid chromatography]. AB - Referring to traditional optimal methods, a method for the optimization of isocratic elution mobile phase composition in high performance liquid chromatography has been developed. In this method, the genetic algorithm based on line-crossover and plane-mutation is used. The principle of genetic algorithm and the process of optimization of mobile phase composition in reversed-phase ion pair high performance liquid chromatography using genetic algorithm are introduced in details. With the concentrations of acetonitrile and ion-pair reagent sodium octane sulfonate chosen as the optimal parameters, the optimum was obtained by three times of optimization procedures. The mean relative error between the predicted and experimental values was 0.75% at the optimum and the optimization results were satisfactory. PMID- 12541962 TI - [Molecularly imprinted polymeric microspheres prepared by seed swelling and suspension polymerization]. AB - A series of molecularly imprinted polymeric microspheres(MIPMs) were prepared by seed swelling and suspension polymerization method in aqueous system using tyrosine as printing molecules, methacrylic acid as functional monomers and trimethylolpropane trimethacrylate (TRIM) as cross-linkers. The morphology including the size, size distribution, pore and pore distribution of the polymer beads was analyzed by scanning electron microscope(SEM). The major factors that influence these properties of the beads are discussed. The molecule selecting property of the MIPMs was detected through liquid chromatography. The results showed that uniform-sized spherical MIPMs had been prepared in aqueous system by seed swelling and suspension polymerization method, and its surface was distributed with pores between 1 micron to 10 microns. The average bead size was adjusted from about 50 microns to 400 microns by changing the stirring speed and the amount of seeds used during the preparation. The prepared MIPMs exhibited a considerable tendency to adsorb tyrosine, the printing molecules selectively, and the separation factor was up to 1.82 when phenylalanine was chosen as the competitive molecules. PMID- 12541963 TI - Endotoxin removal in some medicines and human serum albumin solution by affinity membranes. AB - Three kinds of affinity membranes utilized for endotoxin removal, namely, chitosan affinity membrane (KFCC517), hydrophobic and cationic charged membrane (KFCG316) and metal chelate affinity membrane (KFCM402) were prepared by using cellulose membrane as support. The adsorbability and applications of the membranes in endotoxin removal were investigated. The results showed that all of these affinity membranes could be utilized for endotoxin removal in several kinds of solutions. The factors affecting endotoxin removal such as ionic strength, pH value and flow rate were investigated, and the results were discussed. Three cartridges of membranes KFCC517, KFCG316 and KFCM402 were utilized for endotoxin removal in human serum albumin solution. The cartridge of membrane KFCC517 also was utilized in medicine injections, such as hydrocortisone, puerarin, tetracaine hydrochloride and dextran 40 glucose. Under the optimal conditions, the removal efficiencies of endotoxin were over 80% and the recoveries of desirable substances were over 95%. PMID- 12541964 TI - [Application of electroosmotic pump on micro column liquid chromatography]. AB - An electroosmotic pump(EOP) was designed and evaluated, which could replace the mechanical pump. The EOP could generate 2.0 MPa-6.0 MPa output pressure and tens of nL/min-3 microL/min flow rate. A test mixture containing naphthalene, anthracene and phenanthrene was separated on a 14 cm x 320 microns i.d., 5 microns, C18 micro column with acetonitrile/water as a mobile phase, which demonstrated the applicability of the EOP. PMID- 12541965 TI - [Direct resolution of clenbuterol enantiomers with amide derivative as chiral stationary phase]. AB - A simple and rapid high performance liquid chromatographic method has been developed for the direct resolution of clenbuterol enantiomers. The method involved the use of an amide type chiral stationary phase(CSP) made of (R)-1 naphthylglycine and 3,5-dinitrobenzoic acid known as the Chirex 3005 column. The effects of different contents of n-hexane, 1,2-dichloroethane and methanol in mobile phase on the chiral separation are discussed. The effects of column temperature and flow rate of mobile phase were also studied. The separation factor obtained was 1.32 and the resolution factor was 1.81 when using the optimized mobile phase composed of n-hexane-1,2-dichloroethane-methanol (54:38:8, volume ratio) at 17 degrees C and 1.0 mL/min. The mechanism of separation is also discussed. PMID- 12541966 TI - [Study of the deactivation of the organic-inorganic hybrid sol-gel coatings on stainless steel tubes]. AB - The organic-inorganic hybrid sol-gel coatings used as deactivating substance for stainless steel tubes in gas chromatography have been prepared by dynamic processing of the sol-gel solution produced by hydrolysis and polycondensation of methyltriethoxysilane and tetraethoxysilane under acidic conditions. The chromatographic behavior of the coatings was studied by GC at different aging temperatures. The experimental results indicate that the active sites on the inner surface of the stainless steel tubes have been shielded effectively by the coatings and the coatings also have the ability to separate hydrocarbons. If the coatings were aged only at 250 degrees C, the hydrocarbons with the number of carbon under 12 can be separated with perfect baseline. Compared with the results obtained from the untreated stainless steel tubes the adsorbent activation of the treated stainless steel tubes has decreased greatly. It is also found that when the aging temperature was increased, the action between the coatings and the samples was enhanced, mainly produced by the porous structure of the coatings. This phenomenon is also discussed using the tailing factors. PMID- 12541967 TI - [Capillary electrophoretic analysis of amnesic shellfish toxin--domoic acid]. AB - Domoic acid, a main component of amnesic shellfish toxin (one of the four sorts of marine red tide bio-toxins) was determined by capillary electrophoresis with UV detection. The shellfish samples were prepared by solvent extraction, cleaned up with a strong anion-exchange cartridge (SAX: Part No. 1210-2044, Lot No. 182639, Varian) and a cation-exchange cartridge (SCX: Part No. 1211-3039, Lot No. 171069, Varian). The quantitative analysis was performed with external standard under the optimum conditions of capillary electrophoretic analysis. The calibration curve of domoic acid showed good linearity in the range of 0.2 mg/L 50 mg/L with r = 0.9990, and the detection limit was 0.063 mg/L(S/N > 3). Spiked with standard at three levels (5.00 micrograms/g, 10.00 micrograms/g and 20 micrograms/g), the samples had the recoveries of 97.24%, 96.92% and 97.55%, and the RSDs of 2.74%, 2.59% and 1.95%, respectively. Five kinds of shellfish samples normally consumed in Dalian sea area were collected and quantified. The method has the advantages of being simple, convenient, sensitive and low cost. Therefore it can be used as one of the routine monitoring methods for amnesic shellfish toxin. PMID- 12541968 TI - [Determination of trace level of bromate and perchlorate in drinking water by ion chromatography with microwave preconcentration]. AB - A simple sample preconcentration technique for the determination of trace level of bromate and perchlorate in drinking water with ion chromatography is presented. With the hydrophobic anion-exchange column and a sodium hydroxide eluent in linear gradient, bromate and perchlorate can be determined in a single injection within 35 min. Prior to ion chromatographic analysis, the drinking water sample was treated with the OnGuard Ag cartridge to remove the superfluous chloride and concentrated in 20-fold with poly(tetrafluoroethylene)(PTFE) beaker in a domestic microwave oven in 15 min. The recoveries of the analytes are better than 90%. The detection limits for bromate, perchlorate, iodate and chlorate were 0.10 microgram/L, 0.20 microgram/L, 0.11 microgram/L and 0.21 microgram/L, respectively. The developed method is applicable for the quantitation of bromate and perchlorate in real drinking water samples. PMID- 12541969 TI - [Comparison of the separation performances between semi-preparative chromatographic column and chromatographic pie]. AB - By comparing the resolution and recoveries of proteins between semi-preparative chromatographic column and chromatographic pie (pie-like column) having the same geometric volume under the same chromatographic conditions, the result from the former was found to have the almost same as that form the latter. However, the back pressure of the chromatographic pie rose slower than that of the chromatographic column with the increase of flow rate. Furthermore, the mass and volume loads for the chromatographic pie were higher than those for the chromatographic column. The results indicated that the chromatographic pie had more advantages than the chromatographic column in their performances, and this lay a foundation of the applications of the chromatographic pie in industry. PMID- 12541970 TI - [Simultaneous determination of trace Ni2+, Mn2+, Zn2+ and Cu2+ with microwave derivatization/ion-pair high performance liquid chromatography/meso-tetra(4 sulfophenyl)-porphyrin spectroscopy]. AB - The best conditions for the reaction of meso-tetra(4-sulfophenyl)-porphyrin (TPPS4) with Mn2+, Ni2+, Zn2+ and Cu2+ have been investigated. A method for the fast determination of Ni2+, Mn2+, Zn2+ and Cu2+ has been developed: microwave derivatization at 750 W for 3 min, C18 column, acetonitrile-water(22.5:77.5, volume ratio) as mobile phase with tetraethylammonium bromide (TEABr) as ion-pair reagent and detection wavelength of 415 nm. The detection limits for Zn2+, Cu2+, Ni2+ and Mn2+ were 0.05 microgram/L, 0.01 microgram/L, 0.10 microgram/L and 0.40 microgram/L, respectively. This method has been applied to analyze the concentrations of these four metal ions in marketed tea. PMID- 12541971 TI - [Study of the acupuncture effect on monoamine transmitters in rabbit plasma and brain tissue by high performance liquid chromatography with electrochemical detection]. AB - A rapid and simple method for the study of the acupuncture effect on monoamine transmitters and related compounds in rabbit plasma and brain tissue by high performance liquid chromatography with electrochemical detection was developed. An ODS column was selected as the separation column at 25 degrees C, and pH 4.50, 0.02 mol/L of trisodium citrate-0.05 mol/L sodium phosphate dibasic to methanol (95:5, volume ratio) without ion-pair at a flow rate of 1.0 mL/min. Four compounds, epinephrine (E), norepinephrine (NE), dopamine (DA) and 5 hydroxytryptamine (5-HT), were simultaneously separated and determined under the above conditions. Twenty rabbits were investigated after the acupuncture action upon the central neurotransmitters. The sufficient data showed that acupuncture could significantly affect the activities of the neurotransmitters including E, NE, DA and 5-HT, and the changed functions of the neurotransmitter systems induced by acupuncture not only lead to the neurotransmitter content increase both in brain and plasma but also cause the increase of rabbit breed ability. The results show that the method is very simple and fast. The method is valuable not only for clinical diagnosis but also for research work. PMID- 12541972 TI - [Modification of the analytical method for rotenoids in plants]. AB - Rotenoids are the active ingredients of some botanical insecticides and prospective candidates as anticancer agents. The proper isolation and determination of rotenoids in plants is of great importance for their further research and development. However, the HPLC method available for this purpose was developed particularly for the detection and determination of rotenone, so it appears to be unsuitable for the analysis of other rotenoids such as deguelin, elliptone and their analogues. By checking the UV spectra, it has been found that four types of UV absorption patterns occurred among the major rotenoids isolated from the roots of Derris elliptica and leaves of Tephrosia vogelii, and that the detection wavelength at 240 nm is more adequate for the analysis of a complex of rotenoids than at 280 nm-300 nm, which is used for rotenone detection. The extraction of rotenoids from plants is conveniently carried out by CHCl3 MeOH(9:1, V/V) and the purification can be accomplished by filtration of the crude residue through a C18 reversed-phase cartridge. Rotenoids can be isocratically eluted by MeOH-H2O(66:34, V/V). The results showed that rotenone, deguelin, elliptone, and their 12a-hydroxy- and 6a,12a-dehydro-analogs can be easily detected by the modified method, along with a satisfactory peak separation. The rotenoid components might be characterized by their retention times and relative retention times based on rotenone, which were at a range of 3.26 min-39.42 min and 40.4%-489.1% respectively. PMID- 12541973 TI - [High performance liquid chromatographic determination of internal hormones in inter-subspecific hybrid rice]. AB - A simple, rapid and perfect extraction and determination method for the internal hormones (gibberellin acid (GA3), indole-3-acetic acid (IAA), zeatin (Z), abscisic acid (ABA)) in inter-subspecific hybrid rice with reversed-phase high performance liquid chromatography has been developed. The chromatographic conditions were as follows: a Waters C18 column (4.6 mm i.d. x 250 mm, 5 microns), an SPD-6AV detector set at 254 nm and the mobile phase was a mixture of methanol, water and acetic acid (45:54.2:0.8, volume ratio). The lower detection limits were 0.5 mg/L for GA3, 0.1 mg/L for IAA, 0.3 mg/L for Z and 0.03 mg/L for ABA. PMID- 12541974 TI - Determination of vitamin C in plasma and dialysate from uremia patients by high performance liquid chromatography with electrochemical detection. AB - A convenient and valid method for the determination of ascorbic acid(AA) and dehydroascorbic acid(DHAA) in plasma and dialysate from patients with uremia by high performance liquid chromatography with electrochemical detection is described. A mixture of 0.8 g/L metaphosphoric acid and 18% (volume fraction) perchloric acid was used as a protein precipitant and the extractant for AA from biosamples. It was also a good stabilizer for AA in samples. The proposed method is satisfied for routine screening of vitamin C in clinical applications with a correlation coefficient of more than 0.99 in the range of 2 mumol/L-40 mumol/L AA. The within-day precision was less than 8.9% and 10.55% for AA and DHAA, respectively. The recoveries of AA in plasma and dialysate were over 95% and 78%, respectively. PMID- 12541975 TI - [Direct UV detection of glucose and its derivatives in capillary zone electrophoresis]. AB - UV labeling detection has been commonly used to determine the association constants between lectins and saccharides, but the interaction is always between the labeled carbohydrates, rather than the truly underivatized carbohydrates, and lectins. In order to directly detect saccharides during the study on the interaction of glucose and its derivatives with lectins (e.g., concanavalin A), a capillary zone electrophoretic method with detection at a wavelength of 195 nm has been developed. The influences of various separation conditions including buffer concentration, pH and voltage were investigated. By using an uncoated silica capillary (50 microns i.d., 375 microns o.d., 48.5 cm of total length, and 44.0 cm to the detector) and 50 mmol/L Na2HPO(4)-50 mmol/L NaH2PO4 solution (near to the physiological pH of 7.4) as buffer, the underivatized sugars, including glucosamine, N-acetylglucosamine, glucose, and sodium gluconate, were sufficiently separated within 11 min at an applied voltage of 10 kV. On-column UV monitoring allowed the detection of these compounds at less than 4 mmol/L level, and quantification by the peak area method allowed reproducible determination of them at least at their respective concentration ranges. The method is characterized by its simplicity, rapidity, and reproducibility, and should be useful for the analysis of the interaction of glucose and its derivatives with lectins. PMID- 12541976 TI - Use of capillary electrophoresis with conductivity detection to anions. AB - Capillary electrophoresis with conductivity detection was used to separate and detect chloride (Cl-), nitrate (NO3-), bicarbonate (HCO3-) and dihydric phosphate (H2PO4-) ions. The proposed method was carried out by using citric acid and sodium citrate as buffer solution, under the condition of a-10 kV high voltage power supply. The effects of various buffers, concentrations, pH values and running voltages on separation were investigated. Under the chosen conditions the linear ranges of Cl-, NO3-, HCO3-, and H2PO4- were 5.0 x 10(-5) mol/L -2.5 x 10( 3) mol/L, 6.0 x 10(-5) mol/L-2.0 x 10(-3) mol/L, 5.0 x 10(-6) mol/L-2.0 x 10(-4) mol/L and 6.0 x 10(-5) mol/L-1.0 x 10(-3) mol/L respectively. The detection limits were 1.5 x 10(-5) mol/L, 3.0 x 10(-5) mol/L, 1.0 x 10(-6) mol/L, 2.0 x 10( 5) mol/L and the relative standard deviations (RSD) of migration time were 3.1%, 3.3%, 2.6% and 2.9% respectively. Tap water was analyzed under the same conditions. PMID- 12541977 TI - [Rapid determination of metallothioneins in foods by capillary zone electrophoresis]. AB - A rapid method for the analysis of metallothioneins (MT) in foods by capillary zone electrophoresis(CZE) was developed. Two isomers of MT (MT1, MT2) in foods were separated and determined. After a series of optimization, the separation and determination of MT1 and MT2 were obtained within 10 min by using the phosphate buffer system consisting of 0.02 mol/L Na2HPO4 and 0.02 mol/L NaH2PO4 (pH 7.0), and UV detection at 200 nm. Under the optimal experimental conditions, the minimum detectable limit was 1 mg/L, and the added standard recoveries of MT1, MT2 in foods were found to be in the range of 82.0%-93.4%. The relative standard deviations (RSD) were found to be lower than 10%. Therefore, with this simple and rapid method, the contents of two isomers in foods can be determined by external standard method after sample pretreatment. PMID- 12541978 TI - [Determination of organochlorine pesticides and polychlorinated biphenyl congeners residues in eggs by gas chromatography with electron capture detection (GC-ECD)]. AB - A procedure of clean-up with H2SO4 after extraction with a shaking machine using n-hexane, and fast determination of some organochlorine residues in eggs by GC ECD with PCB2 and PCB209 as internal standards was established. The test results showed that with this method satisfactory recoveries(84.31%-116.77%), relative standard deviation (6%-18%) and detection limits (0.07 ng/g-0.35 ng/g) could be obtained. In comparing with the Soxhlet method, this method is simpler in pre treatment, lower in cost and less in sample amount. It can determine organochlorine pesticides and some PCBs in the same extract, simultaneously with enough accuracy, sensitivity and permissible RSD. It is a suitable procedure for determining micro-amounts of organochlorine pollutants in foods. The results showed that p, p'-DDE was detected in maximum; p, p'-DDT next; and PCBs very little, that were usually less than 10 ng/g in the eggs from some Chinese markets. It was found that most of PCBs in eggs were the congeners with 3-5 chlorine atoms. PMID- 12541979 TI - [Determination of volatile hydrocarbon in atmosphere by two-dimensional capillary column gas chromatography]. AB - Fifty-seven volatile hydrocarbons (VHCs) (C3-C10) in the atmosphere have been determined by two-dimensional capillary column gas chromatography. The samples were preconcentrated using adsorption/two stages of thermal desorption. The samples were analysed by dual columns (PLOT, 50 m x 0.32 mm i.d. x 8 microns, Al2O3/Na2SO4; HP-1, 50 m x 0.22 mm i.d. x 0.5 micron, methyl silicone)/dual flame ionization detectors(FID). The qualitative stability of the method is good. The relative standard deviation of the peak area response is smaller than 20%. The correlation coefficients(r2) for linear regression equations for PLOT and HP-1 columns were 0.9990 and 0.9997 respectively. Analytical period was 48 min. The 182 atmospheric samples from the Ecological Station of Changshu, Jiangsu were measured. The results show that the method is accurate, rapid and practically useful for the determination of the VHCs in atmosphere. PMID- 12541980 TI - [Determination of antioxidants butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) in cosmetics by gas chromatography-mass spectrometry selected ion method]. AB - A new method for the determination of butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) in cosmetics by gas chromatography-mass spectrometry selected ion storage (SIS) method was developed. The BHA and BHT in samples were extracted by methanol. The m/z 165 and m/z 205 were the monitoring ion for BHA and BHT respectively. The detection limits of BHA and BHT in samples were 2.5 micrograms/g and 0.5 microgram/g, respectively. The method is simple, rapid and accurate. PMID- 12541981 TI - [Determination of 7-aminoflunitrazepam, the major metabolite of flunitrazepam in urine by high performance thin-layer chromatography]. AB - A rapid and highly sensitive method, based on high performance thin-layer chromatography, is described for the qualitative and semiquantitative determination of 7-aminoflunitrazepam, the main metabolite of flunitrazepam in human urine. 7-Aminoflunitrazepam in specimen was extracted by solid phase extraction using GDX-403 porous polymer bead as sorbent and ethyl ether as eluant. Fluorescamine was used as a reagent to produce fluorescent product on the plate. The limit of detection was 5 micrograms/L and the limit of quantitation was 15 micrograms/L for 7-aminoflunitrazepam in urine. The method can be successfully used for measuring 7-aminoflunitrazepam in urine samples of the subjects excreted over a 48 h period after receiving 1 mg flunitrazepam orally. The method is applicable to drug examination for the cases of drug-facilitated robbery and rape. PMID- 12541982 TI - [Preparative separation of alkaloids from Cortex Phellodendri by high speed countercurrent chromatography]. AB - High speed countercurrent chromatography was successfully applied to the separation of alkaloids from Cortex Phellodendri with a two-phase solvent system composed of CHCl3-CH3OH-0.5 mol/L HCl (2:1:1, volume ratio). Some alkaloid components were obtained, and identification of berberine was made by thin-layer chromatography. PMID- 12541983 TI - [Determination of benzo[a]pyrene in mainstream smoke by solid-phase extraction and gas chromatography-mass spectrometry]. AB - A novel method is suggested for the determination of benzo[a]pyrene in mainstream smoke. Extraction of Cambridge pads was cleaned up with methanol/n-heptane extraction system and silica solid-phase extraction (SPE). The eluted solution containing benzo[a]pyrene was evaporated gently to complete dryness with N2 flow, then reconstituted in 200 microL ethyl acetate for determination with gas chromatography-mass spectrometry selected ion monitoring (SIM). The proposed method had good results and precision for the separation of pyrene in complex matrices. PMID- 12541984 TI - [Determination of oxadiazon residues in cereals by gas chromatography-mass spectrometry]. AB - An efficient method with microchemical method and solid phase extraction (SPE) technique for the determination of oxadiazon residues in cereals by gas chromatography-mass spectrometry (GC-MS) has been established. The oxadiazon was extracted from the sample with benzene-hexane(1:1, V/V). The extract was cleaned up by a column of 500 mg Supelclean LC-Alumina-N. When the blank rice and corn were spiked with oxadiazon in the range of 0.005 mg/kg-5.00 mg/kg, the recoveries were 90.4%-115.7% for rice and 81.3-109.7% for corn, and with RSD of 2%-6% and 4% 9% respectively. The limit of detection was 0.005 mg/kg. The method is rapid, sensitive, accurate and suitable for the analysis of pesticide residues. PMID- 12541985 TI - Towards an HIV/AIDS health education movement. PMID- 12541986 TI - Project TEACH instructors' handbook. PMID- 12541987 TI - Interview with Paisan Tad-Ud, Founding Chairman of the Thai Network of People Living with HIV/AIDS (TNP+), Bangkok, Thailand. Interview by Karyn Kaplan. PMID- 12541988 TI - HIV education: crossing borders. PMID- 12541989 TI - Diary of a treatment educator. PMID- 12541990 TI - Global treatment update. PMID- 12541991 TI - Reclaiming individual and community power. PMID- 12541992 TI - New research confirms role of heart disease as a treatment by-product. AB - Various antiretroviral drugs lead to metabolic and morphologic problems, according to the body of evidence now available to HIV clinicians for review. However, how HIV physicians should react to this situation when patients have the long-term side effects remains debatable. While discontinuation of certain drugs may be advisable in some cases; in others, the risks may outweigh the benefits of switching regimens. PMID- 12541994 TI - FDA approves new rapid HIV test. AB - The Food and Drug Administration has approved a new rapid HIV diagnostic test kit that provides results with 99.6% accuracy in as little as 20 minutes. PMID- 12541993 TI - AACTG recommendations for metabolic problems. Guide covers insulin resistance and diabetes. AB - The Adult AIDS Clinical Trials Group has responded to research showing a link between HIV antiretroviral treatment and metabolic disorders with guidelines that offer recommendations for assessing, monitoring, and treating the problem. According to the AACTG, up to 40% of HIV patients on a protease inhibitor containing regimen will have impaired glucose tolerance caused by significant insulin resistance, which can lead to increased risk of cardiovascular complication. PMID- 12541995 TI - Atazanavir found to help improve lipid profile. IDSA study offers good news for HIV care. AB - A new protease inhibitor called atazanavir actually was found to improve the lipid profile of HIV patients in a 48-week study. The study compared the total cholesterol from baseline and at 48 weeks following treatment with atazanavir. PMID- 12541996 TI - DAAT may work where DOT model fell short. Good results for drugs issued by methadone clinics. AB - While there are some similarities between HIV and tuberculosis treatments, it's the difference in treatment regimens, duration, and end results that have made HIV clinicians and other somewhat hesitant to use the directly observed treatment strategy, which has been so successful in treating TB, with HIV patients. However, the drawbacks to modeling HIV treatment after DOT have been that unlike TB medication, HIV medication always is prescribed daily and that HIV is incurable so medications must continue for the patient's lifetime, rather than for the six months that a typical TB patient might receive treatment. PMID- 12541997 TI - Efavirenz effects worse than reported, study says. AB - A San Francisco study suggests a greater incidence rate of severe psychiatric illness resulting from HIV treatment with efavirenz than what has previously been reported. PMID- 12541998 TI - FDA notifications. New guidance assists HIV drug development. Changes affect approvals for antiretroviral drugs. AB - The Food and Drug Administration has issued new guidance intended to assist sponsors in the clinical development of drugs for the treatment of HIV infection. PMID- 12541999 TI - Working it out. Scenes from the lives of gay and lesbian youth. PMID- 12542000 TI - In search of solidarity. BP's Gillian Murphy interviews radical AIDS doctor Alan Berkman, physician and firebrand. PMID- 12542001 TI - Mothers living with HIV: setting personal goals and improving family life. PMID- 12542002 TI - Smallpox vaccination and HIV infection. PMID- 12542003 TI - Report from the 4th International Workshop on Adverse Drug Reactions and Lipodystrophy in HIV. PMID- 12542004 TI - The HIV/AIDS vaccine research effort: an update. PMID- 12542005 TI - A promising new anti-HCV protease inhibitor. PMID- 12542006 TI - Current status of antiretroviral treatment interruption and intermittent therapy strategies. PMID- 12542007 TI - World AIDS Day: reflections on the pandemic. PMID- 12542008 TI - 2001 syphilis rates show increase: does this portend a new wave of HIV infection? PMID- 12542009 TI - Which predatory mite can control both a dominant mite pest, Tetranychus urticae, and a latent mite pest, Eotetranychus asiaticus, on strawberry? AB - After biological control of Tetranychus urticae using Phytoseiulus persimilis, a latent mite pest, Eotetranychus asiaticus, was found on strawberries growing in a plastic greenhouse in western Japan. To determine whether the release of P. persimilis, an exotic natural enemy of T. urticae, enhanced the occurrence of the latent pest, the efficiency of P. persimilis in controlling E. asiaticus was compared with the efficiency of two indigenous phytoseiid mites, Amblyseius californicus and A. womersleyi, under laboratory conditions. Phytoseiulus persimilis and A. californicus attacked not only T. urticae but also E. asiaticus. However, the predators produced very few eggs and their offspring rarely reached adulthood when fed on E. asiaticius, so their potential as control agents for the latent mite appears to be low. In contrast, A. womersleyi feeding on E. asiaticus reproduced as well as when fed on T. urticae, and exterminated both species of spider mite. This suggests that A. womersleyi has greater potential as a biological control agent in crops where both species may occur. PMID- 12542010 TI - Carry over effects of dietary crude protein and methimazole in broiler chickens. AB - Seven day old male broiler chickens were fed diets containing 12, 18 or 24% crude protein + 0 or 1 g methimazole/kg diet for 21 days to examine the interaction of the birds' thyroid status and crude protein levels on metabolism. Methimazole (1 methyl-2-mercaptimidazole) inhibits thyroidal production of thyroid hormones and results in hypothyroidism. Birds were fed a diet containing 18% crude protein for an additional 21 days to determine the carry over effects of treatments. Birds were killed at 28 and 49 d. In vitro lipogenesis was inversely related (P < 0.05) to dietary protein levels in control birds at 28 d. Dietary methimazole attenuated (P < 0.05) this effect, resulting in a common rate similar to that attained in the birds fed the highest level of protein without methimazole. Birds fed methimazole for an initial 21-day period (7 to 28 d of age) had greater lipogenic rates (P < 0.05) at 49 d than did their control counterparts. In contrast, methimazole increased (P < 0.05) abdominal fat pad (AFP) lipoprotein lipase (LPL) at both age periods, indicating increased ability by the AFP to remove triglycerides from systemic circulation. Observations at 49 d suggest that perturbations in the thyroid of the young bird may substantially change metabolism in later life. Results also show that obesity in hypothyroid birds cannot be explained by increases in de novo lipogenesis, but probably relates to changes in LPL activity. PMID- 12542011 TI - Established cardiovascular disease and CVD risk factors in a primary care population of middle-aged Irish men and women. AB - Contemporary Irish data on the prevalence of major cardiovascular disease (CVD) risk factors are sparse. The primary aims of this study were (1) to estimate the prevalence of major cardiovascular disease risk factors, including Type 2 Diabetes Mellitus, in the general population of men and women between the ages of 50 and 69 years; and (2) to estimate the proportion of individuals in this age group at high absolute risk of cardiovascular disease events on the basis of pre existing cardiovascular disease or as defined by the Framingham equation. Participants were drawn from the practice lists of 17 general practices in Cork and Kerry using stratified random sampling. A total of 1018 people attended for screening (490 men, 48%) from 1473 who were invited, a response rate of 69.1%. Cardiovascular disease risk factors and glucose intolerance are common in the population of men and women aged between 50 and 69 years. Almost half the participants were overweight and a further quarter met current international criteria for obesity, one of the highest recorded prevalence rates for obesity in a European population sample. Forty per cent of the population reported minimal levels of physical activity and 19% were current cigarette smokers. Approximately half the sample had blood pressure readings consistent with international criteria for the diagnosis of hypertension, but only 38% of these individuals were known to be hypertensive. Eighty per cent of the population sample had a cholesterol concentration in excess of 5 mmol/l. Almost 4% of the population had Type 2 Diabetes Mellitus, of whom 30% were previously undiagnosed. A total of 137 participants (13.5%) had a history or ECG findings consistent with established cardiovascular disease. Of the remaining 881 individuals in the primary prevention population, a total of 20 high-risk individuals (19 male) had a risk of a coronary heart disease event > or = 30% over ten years according to the Framingham risk equation, giving an overall population prevalence of 2.0% (95% CI 1.3 - 3.0). At a risk level > or = 20% over ten years, an additional 91 individuals (8.9%) were identified. Thus a total of 24.4% of the population were at risk either through pre-existing CVD (13.5%) or an estimated 10-year risk exceeding 20% according to the Framingham risk equation (10.9%). Thus a substantial proportion of middle-aged men are at high risk of CVD. The findings emphasise the scale of the CVD epidemic in Ireland and the need for ongoing monitoring of risk factors at the population level and the need to develop preventive strategies at both the clinical and societal level. PMID- 12542012 TI - [Chemotherapy terminology]. PMID- 12542013 TI - [The use of esophagotracheal combination tubes in routine clinical and emergency medicine]. PMID- 12542014 TI - Atomic scale characterization of vacancy ordering in oxygen conducting membranes. AB - This article presents a comprehensive investigation of (La, Sr)FeO3 by correlated atomic resolution annular dark field imaging and electron energy loss spectroscopy. Here, the ability of these techniques to characterize point defect formation and phase transitions under reducing conditions in situ in the scanning transmission electron microscope is evaluated and the influence of oxygen vacancies on the structure-property relationships is discussed. In particular, the evolution of the Ruddlesden-Popper, Brownmillerite, and Aurivillius phases can be associated directly with the ionic and electronic conductivity of the bulk material under different thermodynamic conditions. These results lead naturally to an atomistic defect chemistry model to explain the high temperature ionic and electronic conductivity in this and other perovskite materials. PMID- 12542015 TI - Proceedings of the Workshop on Quantitative Neuroanatomy Tools. Amsterdam, The Netherlands. October 11-12, 2001. PMID- 12542016 TI - Assisted reproductive technologies and insurance under the Americans with Disabilities Act of 1990. PMID- 12542017 TI - The faces behind us. PMID- 12542018 TI - A virtue ethics perspective on research. PMID- 12542019 TI - A new ethic for medicine and society. PMID- 12542020 TI - Metaphysics and Catholic bioethics. PMID- 12542021 TI - Tube feeding and advanced dementia. PMID- 12542022 TI - Living wills and health care proxies. PMID- 12542023 TI - Patients' rights in Europe: where do we stand and where do we go? PMID- 12542024 TI - Health and continued residence: reason or pretext. PMID- 12542025 TI - Consent and capacity in psychiatric practice revisited. PMID- 12542026 TI - The cost and value of new medicines in an era of change. Proceedings of a symposium at the Tufts University European Center. Talloires, France. July 25-27, 2001. PMID- 12542027 TI - Improving spinach, radish, and lettuce growth under red light-emitting diodes (LEDs) with blue light supplementation. AB - Radish (Raphanus sativus L. cv. Cherriette), lettuce (Lactuca sativa L. cv. Waldmann's Green), and spinach (Spinacea oleracea L. cv. Nordic IV) plants were grown under 660-nm red light-emitting diodes (LEDs) and were compared at equal photosynthetic photon flux (PPF) with either plants grown under cool-white fluorescent lamps (CWF) or red LEDs supplemented with 10% (30 micromoles m-2 s-1) blue light (400-500 nm) from blue fluorescent (BF) lamps. At 21 days after planting (DAP), leaf photosynthetic rates and stomatal conductance were greater for plants grown under CWF light than for those grown under red LEDs, with or without supplemental blue light. At harvest (21 DAP), total dry-weight accumulation was significantly lower for all species tested when grown under red LEDs alone than when grown under CWF light or red LEDs + 10% BF light. Moreover, total dry weight for radish and spinach was significantly lower under red LEDs + 10% BF than under CWF light, suggesting that addition of blue light to the red LEDs was still insufficient for achieving maximal growth for these crops. PMID- 12542028 TI - Toward legal recognition for termination of life without request? Remarks on Advice No. 9 of the Belgian Advisory Committee on Bioethics concerning termination of life of incompetent patients. PMID- 12542029 TI - The intrepid ethicist: Peter Singer remains undeterred in putting forth his ground-breaking ideas. Interview by Norm Allen. PMID- 12542030 TI - [Re: Kay Brune. Altex 19, 3, 2002: Animal experimentation in sciences: sadistic nonsense or indispensible necessity?]. PMID- 12542031 TI - CRG launches genetic bill of rights. PMID- 12542032 TI - A geostatistical analysis of small-scale spatial variability in bacterial abundance and community structure in salt marsh creek bank sediments. AB - Small-scale variations in bacterial abundance and community structure were examined in salt marsh sediments from Virginia's eastern shore. Samples were collected at 5 cm intervals (horizontally) along a 50 cm elevation gradient, over a 215 cm horizontal transect. For each sample, bacterial abundance was determined using acridine orange direct counts and community structure was analyzed using randomly amplified polymorphic DNA fingerprinting of whole-community DNA extracts. A geostatistical analysis was used to determine the degree of spatial autocorrelation among the samples, for each variable and each direction (horizontal and vertical). The proportion of variance in bacterial abundance that could be accounted for by the spatial model was quite high (vertical: 60%, horizontal: 73%); significant autocorrelation was found among samples separated by 25 cm in the vertical direction and up to 115 cm horizontally. In contrast, most of the variability in community structure was not accounted for by simply considering the spatial separation of samples (vertical: 11%, horizontal: 22%), and must reflect variability from other parameters (e.g., variation at other spatial scales, experimental error, or environmental heterogeneity). Microbial community patch size based upon overall similarity in community structure varied between 17 cm (vertical) and 35 cm (horizontal). Overall, variability due to horizontal position (distance from the creek bank) was much smaller than that due to vertical position (elevation) for both community properties assayed. This suggests that processes more correlated with elevation (e.g., drainage and redox potential) vary at a smaller scale (therefore producing smaller patch sizes) than processes controlled by distance from the creek bank. PMID- 12542033 TI - Introduction to research ethics. PMID- 12542034 TI - [Re: Kay Brune. Altex 19, 3, 2002: Animal experimentation in sciences: sadistic nonsense or indispensible necessity?]. PMID- 12542035 TI - A tailored pelvic floor exercise program commenced immediately post-partum promotes continence. PMID- 12542036 TI - Community physiotherapy provides a small transient benefit for stroke patients with long term mobility problems. PMID- 12542038 TI - Exercise-based cardiac rehabilitation is effective in reducing cardiac deaths. PMID- 12542037 TI - [The ESAC point of view]. PMID- 12542039 TI - Ottawa Ankle Rules are more sensitive than Dutch in detecting significant ankle fracture. PMID- 12542040 TI - A multidisciplinary evaluation and management unit reduces mortality in sick, frail older patients. PMID- 12542041 TI - Placebos in clinical trials: new forms of an old issue. PMID- 12542042 TI - Effects of dipfluzine on delayed afterdepolarizations and triggered activity induced by ouabain in guinea pig papillary muscles. AB - AIM: To investigate the effects of dipfluzine (Dip) on delayed afterdepolarizations (DADs) and triggered activity (TA) induced by ouabain and high Ca2+ in guinea pig papillary muscles. METHODS: Stable and reproducible DADs and TA in guinea pig papillary muscles were induced by ouabain (1 micromol/L) and high Ca2+ (5.4 mmol/L). DADs and TA were recorded using intracellular glass microelectrode technique. RESULTS: (1) DADs and TA were markedly inhibited by pretreatment with Dip (10, 30 micromol/L). The amplitude and duration of DADs were reduced by Dip (30 micromol/L) from 10.5 mV +/- 2.2 mV and 230 ms +/- 19 ms to 3.6 mV +/- 0.3 mV and 152 ms +/- 14 ms, respectively, and the induced time of DADs was prolonged from (21+/-5) to (66+/-11) min. TA was not observed. (2) Dip (10, 30 micromol/L) had significant therapeutic effects on DADs and TA. The amplitude and duration of DADs were reduced by Dip (30 micromol/L) from 10.4 mV +/- 1.2 mV and 218 ms +/- 22 ms to 3.3 mV +/- 0.6 mV and 159 ms+/-26 ms. The occurrence of TA was also abolished. CONCLUSION: Dip has inhibitory effects on DADs and TA induced by ouabain and high Ca2+ in guinea pig papillary muscles, which might be related to alleviation of intracellular calcium overload through inhibiting calcium channel and/or calcium release from sarcoplasmic reticulum. The effects of Dip on DADs and TA might produce anti-arrhythmic effects. PMID- 12542043 TI - Human subjects issues in Internet research. PMID- 12542044 TI - Ethical issues in migrant farmworker studies. PMID- 12542045 TI - Prisoner participation in biomedical research. PMID- 12542046 TI - Protecting human research subjects: ten common problems. PMID- 12542047 TI - Family and community in genetic research. PMID- 12542048 TI - Filming in the ER: invasion of privacy or public service? PMID- 12542049 TI - Levinas and the euthanasia debate. AB - The philosophers' tendency to characterize euthanasia in terms of either the right or the responsibility to die is, in some ways, problematic. Stepping outside of the analytic framework, the author draws out the implications of the ethics of Emmanuel Levinas for the euthanasia debate, tracing the ways Levinas' position differs not only from the philosophical consensus but also from the theological one. The article shows that, according to Levinas, there is no ethical case for suicide or assisted suicide. Death cannot be assumed or chosen- not only because suicide is a logically and metaphysically contradictory concept but also because in the choice of death ethical responsibility turns into irresponsibility. However, since Levinas holds that one must be responsible to the point of expiation, he can be said to approve certain actions that may have the consequence of hastening death. PMID- 12542050 TI - Dysfunction of myocardial taurine transport and effect of taurine supplement in rats with isoproterenol-induced myocardial injury. AB - AIM: To study the alterations of myocardial taurine transport function, taurine transporter (TAUT), and cysteine sulfinate decarboxylase (CSD) mRNA as well as effect of exogenous taurine in rats with isoproterenol (ISO)-induced cardiomegaly. METHODS: [3H]-Taurine uptake and release on myocardium were determined. Binding sites of [3H]-taurine for myocardial sarcolemma were measured. TAUT and CSD mRNA levels were assayed using competitive quantitative reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: ISO group as compared with control group, myocardial taurine uptake markedly reduced, taurine release obviously increased; Bmax value of [3H]-taurine binding on cardiac sarcolemma reduced by 42% (P<0.05); TAUT and CSD mRNA levels decreased by 40% and 38% (P<0.05), respectively. ISO+taurine group as compared with ISO-treated group, the amounts of taurine uptake and TAUT mRNA returned to normal; taurine release reduced; Bmax increased by 92% (P<0.01), and CSD mRNA content augmented by 23% (P<0.05). There were no statistical differences of Kd values among the four groups (P>0.05). CONCLUSION: The data indicate that the failure to generate sufficient TAUT on myocardial sarcolemma may be the fundamental abnormality in ISO-induced cardiac injury. The mechanism by which administration of taurine considerably improves ISO-induced cardiac damage is probably to increase the expression of TAUT gene and recover taurine transport function. PMID- 12542051 TI - Ethical issues of animal welfare in Jewish thought. PMID- 12542052 TI - The right-to-die movement: extrapolating from the National Hemlock Society U.S.A. membership survey. AB - A national membership survey of Hemlock Society USA was conducted by Fox and Kamakahi (1995). Respondents (N=6398) were asked a variety of questions, but in this paper we perform a longitudinal analysis of the characteristics of Hemlock Society USA members. Hemlock Society USA members are divided into three 5-year cohorts: Early Joiners (11 or more years of membership), Middle Joiners (6 to 10 years membership), and Late Joiners (5 or fewer years of membership). Differences between cohorts are examined and extrapolations made regarding Hemlock Society USA and the Right-to-Die Movement. A series of one-way ANOVAs were used with Scheme post-hoc comparisons as heuristic tools for assessing between-cohort differences. Late Joiners are different from earlier members, but are more like other Hemlock Society USA members than the adult U.S. population at large. Hemlock Society USA members are essentially societal "elites" (based on socio demographic variables) who work in social environments that are decidedly split on the issue of voluntary suicide and euthanasia. PMID- 12542054 TI - Abortion and the Supreme Court: advancing the culture of death. PMID- 12542053 TI - 4,4,4-trifluoro-3-(indole-3-)butyric acid promotes root elongation in Lactuca sativa independent of ethylene synthesis and pH. AB - We studied the mode of action of 4,4,4-trifluoro-3- (indole-3-) butyric acid (TFIBA), a recently described root growth stimulator, on primary root growth of Lactuca sativa L. seedlings. TFIBA (100 micromoles) promoted elongation of primary roots by 40% in 72 h but inhibited hypocotyl growth by 35%. TFIBA induced root growth was independent of pH. TFIBA did not affect ethylene production, but reduced the inhibitory effect of ethylene on root elongation. TFIBA promoted root growth even in the presence of the ethylene biosynthesis inhibitor L-alpha-(2 aminoethoxyvinyl)glycine. TFIBA and the ethylene-binding inhibitor silver thiosulphate (STS) had a similar effect on root elongation. The results indicate that TFIBA-stimulated root elongation was neither pH-dependent nor related to inhibition of ethylene synthesis, but was possibly related to ethylene action. PMID- 12542056 TI - Racism: a central problem for the Human Genome Diversity Project. PMID- 12542055 TI - Further commentary on Fox's "In the Matter of 'Baby M'". PMID- 12542057 TI - Ethics in split treatment. PMID- 12542058 TI - Confidential! Genetics research is prompting calls for new privacy laws--before it's too late. PMID- 12542059 TI - Hypolipidemic treatment in children and young adults. Proceedings of a workshop of the International Task Force for Prevention of Coronary Heart Disease. VIlla Orlandi, Italy. May 19, 2001. PMID- 12542061 TI - Thrombolytic therapy in patients with submassive pulmonary embolism. PMID- 12542062 TI - Thrombolytic therapy in patients with submassive pulmonary embolism. PMID- 12542063 TI - Thrombolytic therapy in patients with submassive pulmonary embolism. PMID- 12542064 TI - Thrombolytic therapy in patients with submassive pulmonary embolism. PMID- 12542065 TI - Inhibition of tumor necrosis factor alpha and ankylosing spondylitis. PMID- 12542066 TI - Helicobacter pylori. PMID- 12542067 TI - Changing health insurance trends. PMID- 12542068 TI - XXIX Annual Meeting on Basic Research in Chagas Disease and XVIII Meeting of the Brazilian Society of Protozoology. Caxambu, Brasil, 4-6 November 2002. Abstracts. PMID- 12542069 TI - [Abstracts of the International Meeting of Cardiology. Paris, France, May 27-28, 1999]. PMID- 12542071 TI - Allergen immunotherapy: a practice parameter. American Academy of Allergy, Asthma and Immunology. American College of Allergy, Asthma and Immunology. PMID- 12542073 TI - [Pathology 2002 Meeting. 4-8 November 2002, Paris, France. Proceedings and abstracts]. PMID- 12542074 TI - National Asthma Education and Prevention Program. Expert Panel Report: Guidelines for the Diagnosis and Management of Asthma Update on Selected Topics--2002. PMID- 12542075 TI - What is driving health care cost increases? PMID- 12542076 TI - Converging on nursing home quality. AB - This paper looks at nursing home quality initiatives, built around public reporting of quality data, that have been inaugurated by the Centers for Medicare and Medicaid Services and the California Health Care Foundation. How the projects were developed is explored, along with preliminary indicators of their impact on consumers and providers and likely next steps in their evolution. PMID- 12542077 TI - Back in the driver's seat: specialists regaining autonomy. AB - Between 1997 and 2001, the proportion of specialists reporting more freedom to make clinical decisions that meet their patients' needs increased significantly, much more so than among primary care physicians (PCPs), according to a new study by the Center for Studying Health System Change (HSC). Specialists now are also more likely to believe they can make clinical decisions in the best interest of their patients without reducing their income and can maintain continuing relationships with their patients to promote high-quality care. In contrast, PCPs' views on these issues have changed little. These findings about physicians' perceptions are likely a reflection of recent changes in managed care. Responding to a strong consumer and physician backlash, health plans gave consumers a broader choice of physicians and eased restrictions on care in the late 1990s PMID- 12542078 TI - State eligibility rules under separate state SCHIP programs--implications for children's access to health care. AB - This Policy Brief is the fourth in a series of reports issued by the George Washington University Center for Health Services Research and Policy that examine the design of separately-administered State Children's Health Insurance Programs (SCHIP) that is, programs that operate directly under the authority of the federal SCHIP statute rather than expansions of state Medicaid programs. These Policy Briefs also consider the implications of states' design choices for children's access to health care. The first three briefs in this series focused on three aspects of separate SCHIP programs: children's legal right to assistance under separate programs; benefit and coverage design choices under SCHIP plans; and the design and structure of freestanding managed care contracts negotiated by SCHIP agencies. This issue brief focuses on how financial eligibility for SCHIP actually is calculated, that is, the formulas that states have developed to count children's family income for purposes of measuring eligibility. This topic is of central importance to overall program administration because of the federal legal prohibition against assistance to targeted low-income children who are in fact Medicaid-eligible. This prohibition on duplication of assistance was a crucial assumption in the enactment of SCHIP. It is also key to the conservation of limited SCHIP funding for targeted low-income children who are ineligible for either Medicaid or any other form of health insurance, particularly as unemployment rises and the number of lower income children without health insurance may be poised to increase. PMID- 12542079 TI - Behavioral health and managed care contracting under SCHIP. AB - This Policy Brief examines behavioral health managed care contracting under separately administered State Children's Health Insurance Programs (SCHIP), i.e., programs that operate under the direct authority of Title XXI of the Social Security Act rather than as expansions of Medicaid. Most separate SCHIP programs buy managed care style health insurance for some or most of their enrolled children. Because Title XXI provides states with far greater administrative flexibility than Medicaid with respect to coverage and benefit design, provision of services, and administration of managed care arrangements,studying separate SCHIP managed care products sheds important light on how states might approach insurance and managed care design generally in the area of behavioral health were Medicaid modified through section 1115 demonstration or federal statutory authority to permit greater latitude. To conduct this analysis, two nationwide databases maintained by the George Washington University Center for Health Services Research and Policy (CHSRP) were used: a database consisting of all Medicaid MCO-style managed care contracts in use in Calendar Year 2000; and a nationwide database consisting of contracts used by separate SCHIP programs for the same calendar year. As of the point of collection in 2000 there were 33 such separate programs; according to CMS' latest website information, that total has now reached 35. Both sets of contracts were analyzed and separated into their components by lawyers experienced in managed care contract analysis and interpretation. The data were entered into working tables that organize the contents of the contracts into a series of searchable domains. PMID- 12542080 TI - The next frontier: vascular ultrasound. PMID- 12542081 TI - Awareness of dying: an experience of Chinese patients with terminal cancer. AB - Despite the effort to provide an alternate way to dying, there seems to be little effort to examine the real situation of hospice care. For example, the patients' desired outcomes are seldom addressed. It is recently that studies began to investigate the features of "good death." However, work in eastern cultures, such as Chinese, remains scarce. The purpose of this study is to gain an understanding of what it means to die a "good death" from the perspective of Chinese patients. Thirty-three Chinese hospice patients with terminal cancer were interviewed. Grounded in the analysis of qualitative data, seven elements that contribute to dying a good death emerged. Awareness of dying was identified as the foremost essential element of a good death. From the findings of this study, most respondents considered receiving a diagnosis of cancer as one of the significant points of psychological distress in the course of their illness. Half of the patients openly talked about their cancers and more than half of them discussed various issues related to death. That means, about one-third of the respondents talked about death and cancer with openness. These preliminary findings may reflect the progress of hospice work in Hong Kong. Meanwhile, 13 respondents did not mention the word "death" and 16 of them did not mention that they had cancer. They talked about their illness and future using the expressions that they preferred. It is important that health care professionals also respect people who show awareness of dying as much as those who do not present clear evidence of this awareness. Both groups of people can have a positive experience when they die. PMID- 12542082 TI - Traube's double tone. Invited comment on: "Reports from the clinic of Privy Councilor Traube: two peculiar phenomena regarding aortic insufficiency". PMID- 12542083 TI - ICN announces its position on advanced nursing roles. PMID- 12542084 TI - INR author speaks out: lack of data inhibits efforts to combat nursing shortages. PMID- 12542085 TI - Hope springs eternal. Interview by Alastair McLellan. PMID- 12542086 TI - Cloning and characterization of Edwardsiella ictaluri proteins expressed and recognized by the channel catfish Ictalurus punctatus immune response during infection. AB - An Edwardsiella ictaluri expression library was screened for clones expressing antigenic E. ictaluri proteins using anti-E. ictaluri serum, which resulted in the isolation of 32 clones. The clones were partially characterized and 4 were selected for complete analysis. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), 2-dimensional PAGE, Western blotting, and DNA sequencing were used to analyze expressed antigenic proteins and encoded genes. Sequence analysis identified 4 putative open reading frames (ORFs) in the insert of Clone 4d6, which corresponded to antigenic acidic proteins of 55, 20 and 18 kDa expressed by both the clone and E. ictaluri cells. The predicted gene products of these ORFs were similar to several products of the imp locus of Rhizobium leguminosarum bv. trifolii. The imp locus of R. leguminosarum contains 14 genes that encode proteins involved in a putative temperature-dependent protein secretion system. In addition there was significant amino acid identity for a variety of hypothetical proteins from R. solanacearum, Ps. aeruginosa, A. tumefaciens, Y. pestis, and Salmonella typhimurium. Overlapping inserts of Clones 1.4, 5d2, and 5d3 encoded ORFs similar to Escherichia coli partial genes serA and pgk, and complete genes rpiA, iciA, yggE, yggB and fda. These genes encode D-3 phosphoglycerate dehydrogenase (serA), ribose 5-phosphate isomerase (rpiA), a specific inhibitor of chromosomal initiation of replication (iciA), a hypothetical protein (yggE), a protein involved in responses to osmotic stress (yggB), fructose 1,6-bisphosphate aldolase (fda), and phosphoglycerate kinase (pgk). Cloned antigenic E. ictaluri proteins of 33, 27, 35 and 45 kDa appeared to be products of the ORFs similar to yggE, rpiA, iciA, and fda respectively. All the cloned antigenic proteins were recognized by antiserum from catfish that had recovered from enteric septicemia of catfish (ESC), indicating that these antigens are expressed during the infectious process. The cloned antigenic proteins were subsequently evaluated as subunit vaccines for protection against wild-type E. ictaluri. All vaccine treatments were protective against E. ictaluri in catfish, but results were inconclusive due to high levels of cross-reactive protection afforded by the E. coli host strain of the cloning vector. PMID- 12542087 TI - Occurrence of Flavobacterium psychrophilum in fish-farming environments. AB - Occurrence of Flavobacterium psychrophilum in fish farms and fish-farming environments was studied using agar plate cultivation, the immunoflourescence antibody technique (IFAT) and nested PCR. Characteristics of 64 F. psychrophilum isolates from rainbow trout Oncorhynchus mykiss, fish farm rearing water, ovarian fluid and wild fish were serotyped, ribotyped and compared biochemically. Virulence of F. psychrophilum isolates from different sources was compared by injection into rainbow trout. Additionally, the number of F. psychrophilum cells shed by naturally infected rainbow trout was determined. F. psychrophilum was detected and isolated from skin mucus, skin lesions and internal organs of diseased rainbow trout and from fish without clinical disease. The pathogen was also present in wild perch Perca fluviatilis, roach Rutilus rutilus, and ovarian fluids of farmed rainbow trout brood fish. Isolates were biochemically homogenous, excluding the capability to degrade elastin. Five different agglutination patterns with different antisera against F. psychrophilum were found among the isolates studied. Although several different ribopatterns were found (ClaI: 12 ribopatterns and HaeIII: 9 ribopatterns), ribotype A was the most dominant. Farmed rainbow trout brood fish carried a broad-spectrum of serologically and genetically different F. psychrophilum in ovarian fluids. Virulence of the tested isolates in rainbow trout varied and naturally infected rainbow trout shed 10(4) to 10(8) cells fish(-1) h(-1) of F. psychrophilum into the surrounding water. PMID- 12542088 TI - Inhibition of chemiluminescent response of olive flounder Paralichthys olivaceus phagocytes by the scuticociliate parasite Uronema marinum. AB - Experiments were conducted to evaluate the in vitro capacity of the scuticociliatian parasite Uronema marinum to inhibit chemiluminescence (CL) of olive flounder Paralichthys olivaceus phagocytes. Luminol-enhanced CL was used to measure the production of reactive oxygen intermediates (ROIs) generated by respiratory bursts of phagocytes using zymosan as a stimulant. Cytotoxic and antioxidative activities of excretory-secretory (ES) products of the parasite were evaluated as well. Live U. marinum and its ES products had a negative and dose-dependent effect on luminol-enhanced CL responses of zymosan-stimulated phagocytes of olive flounder. After CL assay, the number of phagocytes showing viability was significantly reduced in the cells incubated with live U. marinum at ratios of 2:1 and 1:1 phagocytes:ciliates or ES products with 0.3 mg protein ml(-1) compared to controls. Lysis of phagocytes by exposure to ES products was observed also. ES products from U. marinum showed considerably high activities of superoxide dismutase (SOD) and catalase. The results of this study suggest that U. marinum can protect itself against host's phagocytes mediated oxidative damage by destroying phagocytes and scavenging ROIs. PMID- 12542089 TI - Site preference of myxosporean spp. on the fins of some Hungarian fish species. AB - Some species of the known fish myxosporeans select fins as type location for their development. During the plasmodial development of Myxobolus, Henneguya and Thelohanellus species in cyprinid fishes and eels, 3 main locations of plasmodial development were distinguished inside the fins. M. portucalensis, M. alburni, M. caudatus and a Myxobolus sp. formed plasmodia in the dermis of the skin doublets between finrays, while plasmodia of M. diversus and H. cutanea developed inside the lumen of the cartilaginous finrays. The third type of development was represented by T. nikolskii, which started its development on the outer surface of the finrays. Plasmodia both in the lumen and on the surface of the finrays were surrounded by a cartilaginous capsule, but plasmodia of T. nikolskii also had a thick connective tissue capsule containing pieces of cartilage. PMID- 12542090 TI - Dormancy as a survival strategy of the fish pathogen Streptococcus parauberis in the marine environment. AB - The fate of Streptococcus parauberis in seawater and sediment microcosms at different temperatures (6 and 22 degrees C) was investigated by comparing the survival dynamics of 2 strains of this bacterial species, isolated respectively from diseased turbot and cattle. The turbot and the bovine isolate showed similar survival kinetics, remaining culturable for approximately 1 mo in water and 6 mo in sediment. A slight influence of temperature on the stability of the cells was observed, in that the number of culturable cells was about 1 log10 unit higher at 6 than at 22 degrees C. During the starvation period, the metabolic activity of the cells, after suffering a strong reduction during the first 12 d, stabilized at levels ranging from 20 to 40% of the initial values. However, in all the microcosms, the acridine orange (AO) and 4',6-diamidino-2-phenilindole (DAPI) counts remained at about 10(5) cells ml(-1) throughout the experimental period, even when cells became undetectable by standard plate count methods. The addition of fresh medium to microcosms containing nonculturable cells induced the return to culturability of S. parauberis strains. On the basis of these results, it seems that S. parauberis has the ability to enter into a viable but nonculturable (VBNC) state. Dormant cells of the turbot isolate maintained their infectivity and pathogenic potential for fish. PMID- 12542091 TI - Cockle emergence at the sediment surface: 'favourization' mechanism by digenean parasites? AB - The aim of the present work was to assess the effect of digenean trematodes on indirect mortality of the cockle Cerastoderma edule, an infaunal bivalve. The tested hypothesis was that parasites altered the burrowing capacity of cockles and thus exposed them at the sediment surface, where they are more vulnerable to predators. If the predator is the final host, this mechanism, which drives the cockle out of the sediment, is considered as a 'favourization'. Cockle populations from 2 stations in Arcachon Bay (France)-Banc d'Arguin (oceanic situation) and La Canelette (lagoonal situation)--were sampled for 1 yr. At La Canelette, monitoring every 2 d showed that 50% of adult cockles regularly migrated to the sediment surface at a rate of 5 cockles m(-2) yr(-1) and disappeared in a few days. In the laboratory, 67% of these 'surface cockles' did not burrow again, suggesting that they would die in the field. Moreover, mortality measured after 7 d in the laboratory was 2 to 5 times higher than mortality of 'buried cockles', at both stations and particularly during summer. Species richness and abundance of digeneans from both stations were compared in 'buried' and 'surface' individuals to determine whether parasites played a role in cockle migration and mortality. Ten and 9 digeneans were found at Banc d'Arguin and La Canelette, respectively, with Himasthla quissetensis and Labratrema minimus being the most prevalent and abundant species at both stations. The abundance of H. quissetensis was slightly higher in surface cockles at Banc d'Arguin, but the difference fluctuated with station and cockle age (or size). L. minimus prevalence was only higher in surface cockles at La Canelette. In the latter station, we estimated that L. minimus and H. quissetensis were responsible for the emergence of 9 and 2%, respectively, of the buried cockles. Although this favourization mechanism may induce some mortality in cockles, it does not alone explain the magnitude of the observed mortalities (41 and 57% at La Canelette and Banc d'Arguin, respectively). A correspondence analysis did not show the presence of a particular parasite community in buried or surface cockles, which could explain these high surface cockle mortalities in association with the 2 dominant digeneans. PMID- 12542092 TI - Dynamics of shell disease in the edible crab Cancer pagurus: a comparative study between two sites on the Gower Peninsula, South Wales, UK. AB - Shell disease syndrome is a degradative condition of the crustacean exoskeleton which results in the formation of black-spot lesions. Field surveys on the edible crab Cancer pagurus (L.) population at 2 sites in the Gower Peninsula of South Wales, UK, namely Langland Bay and Rhossili causeway, revealed similarities in the prevalence of shell disease but differences in the severity. Male crabs from Langland Bay displayed significantly greater severities of the disease on the dorsal carapace and ventral surfaces than those in Rhossili. The greatest severities of the disease in Langland were attributable to sand abrasion injuries during back-burrowing behaviour and prolonged exposure to high numbers of chitinolytic bacteria. Lower severities of shell disease in males from Rhossili were a reflection of the inability to back-burrow due to the nature of the onshore sediment, and hence the crabs were only exposed to low levels of chitinolytic bacteria in the water column. On areas of the causeway that permitted back-burrowing, the chitinase enzyme activities in sediments were significantly lower than those in Langland Bay. It is concluded that the potential for crustacean shell degradation is higher at Langland than Rhossili, but whether the differences in chitinolytic abilities of the sediments at the 2 sites are a result of differences in pollution levels or simply of different substratum types, remains to be determined. PMID- 12542093 TI - Transmission of crayfish plague. AB - Two possible means of transmission of crayfish plague were investigated: via fish (as vectors), and via crayfish (as hosts or vectors when dead). The crayfish transmission experiments focussed on both the viability of the fungus in dead crayfish when kept in simulated field conditions, and on the treatments which kill viable forms of Aphanomyces astaci within the recently dead host (cadaver). It was found that A. astaci remains viable for 5 d, and possibly longer in crayfish kept in water at 21 degrees C after dying of crayfish plague. Heating (boiling for 1 min) was the quickest way of decontaminating crayfish cadavers. Freezing took considerably longer: after 48 h at -20 degrees C, viable stages were still present. It was also found that A. astaci is unlikely to survive passage of the gastrointestinal tract of either mammals or birds as no viable stages were found after 12 h at 37 degrees C. Two basic modes of transmission of crayfish plague via fish were investigated: (1) after passage of initially viable forms of A. astaci through the digestive tract of fish and (2) via fish skin. If A. astaci was fed to fish as infected abdominal cuticle, it was still viable after passage through the gastrointestinal tract. When pure mycelium or spores were fed to fish there was no indication of viable forms of A. astaci after passage through the gastrointestinal tract. Transmission via fish skin was not observed under the experimental conditions applied. The results on fish as vectors have practical importance for fish transport and stocking because the present study shows that there is a risk of transmission of crayfish plague via fish faeces. The investigation of crayfish as vectors delivers methods that could be used for treatment of crayfish imported for human consumption into crayfish plague-free areas. This application is particularly important for the importation of American crayfish, which are in general suspected to carry A. astaci in their cuticle. PMID- 12542094 TI - Parasitic infections in live freshwater tropical fishes imported to Korea. AB - We examined 15 species of ornamental tropical fishes originating from Southeast Asia to determine the cause of losses among 8 fish farms in Korea. A total of 351 individuals belonging to 5 different families (1 species of Characidae, 6 of Cichlidae, 3 of Cyprinidae, 1 of Heleostomatidae, and 4 of Poecilidae) were collected for the purpose of detecting metazoan and protozoan parasites. Parasites were fixed and stained using routine methods, and identified. We found 3 ciliates, 2 monogeneans, 1 nematode, and 1 copepod from 7 host species. Of these, Ichthyophthirius multifiliis was the most common parasite in our study, and together with Trichodina sp., caused mass mortality of Sumatra barb Puntius tetrazona at 1 farm. We also found Camallanus cotti and Tetrahymena corlissi from guppies Poecilia reticulata, both for the first time in Korea. Farmers consider these 2 pathogens to be the most serious ones in Korea. Gussevia asota from oscar Astronotus ocellatus, and Gyrodactylus bullatarudis from platy Xiphophorus maculatus were also found in Korea for the first time. We believe that appropriate quarantine practices for tropical ornamental fishes should be introduced because the failure to require and implement quarantines has already resulted in the accidental introduction of exotic parasites to fish farms, and because these parasites can cause further economic losses if they become established in the wild. PMID- 12542095 TI - Detection of the parasitic dinoflagellate Hematodinium in the Norway lobster Nephrops norvegicus by ELISA. AB - Norway lobsters Nephrops norvegicus from the coastal waters of Scotland are seasonally infected by a parasitic dinoflagellate of the genus Hematodinium. An enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of the parasite in the haemolymph of N. norvegicus. The ELISA is simple to perform with a detection limit of 5 x 10(4) parasites ml(-1) haemolymph. The ELISA is currently being used to study the prevalence and seasonality of Hematodinium infection in N. norvegicus and other crustacean hosts. PMID- 12542096 TI - It works for me! PMID- 12542097 TI - Glass ionomers. PMID- 12542098 TI - Diagnosis and management of maxillary incisors affected by incisal wear: an interdisciplinary case report. AB - In the attempt to restore anterior teeth affected by erosion and bruxism, many clinicians have been frustrated with the constant restorative failures. Frequently, these failures are attributed to the restorative materials employed, especially in cases in which composite resins are used. However, some flaws of the restorations are related to the oversight of occlusal principles. The purpose of this article is to discuss the etiology, signs, and symptoms of incisal wear, with special attention to that caused by bruxism and chemical erosion. Relatively simple management techniques (e.g., occlusal adjustment, adhesive restorations) are proposed, and the diagnosis and management of a representative clinical case is presented. CLINICAL SIGNIFICANCE: In some cases of bruxism and/or dental erosion, it is possible to acquire space to recuperate the esthetics and function of maxillary incisors affected by incisal wear through a conservative treatment associated with the control of the etiologic factors. PMID- 12542099 TI - Esthetic rehabilitation of anterior teeth affected by enamel hypoplasia: a case report. AB - Alterations during tooth structure formation, mainly on anterior teeth, are known to severely compromise esthetics. This article presents a case report in which the etiology and clinical characteristics of anterior teeth affected by enamel hypoplasia are analyzed. The restoration treatment sequence is also discussed, demonstrating that the use of a hybrid and a microfilled composite associated with tooth hybridization (interdiffusing zone) can be used to conservatively rehabilitate a smile. CLINICAL SIGNIFICANCE: This article presents a simple and conservative technique for restoring disfiguring areas of enamel hypoplasia to achieve an optimally esthetic result. PMID- 12542100 TI - Capping the inflamed pulp under different clinical conditions. AB - BACKGROUND: A great deal of controversy exists regarding the reliability of capping the inflamed pulp. In particular, the use of calcium hydroxide as a capping agent has come into question. In this study, hard tissue barrier formation after inflamed pulps were capped directly or after partial pulpotomy was compared with calcium hydroxide or bonded resin and with no additional seal or an IRM surface seal. Seventy teeth in five dogs were used. Ten untreated teeth were used as negative controls. In 60 teeth, pulpal inflammation was induced by preparing a cavity close to the pulp and sealing a cotton pellet soaked in plaque in it for 1 to 2 weeks. The cavities were then re-entered and extended to expose the pulps. MATERIALS AND METHODS: In half the teeth (n = 30) a partial pulpotomy was performed and in the other half (n = 30) pulpal treatment was performed on the superficial exposed pulp. Both pulpal treatment groups received the same restorative procedures: (1) calcium hydroxide + amalgam + IRM surface seal; (2) OptiBond Solo, Prodigy with IRM surface seal; or (3) OptiBond Solo, Prodigy without IRM surface seal. The presence, absence, and quality of a hard tissue barrier were evaluated histologically. RESULTS: The calcium hydroxide groups were statistically superior to all other groups. The IRM surface seal resulted in significantly better healing. Although there was no statistically significant difference between direct pulp capping and partial pulpotomy with the numbers in this study, power statistics indicated that in clinical practice a partial pulpotomy would be preferable. CLINICAL SIGNIFICANCE The results of this study suggest that a partial pulpotomy, calcium hydroxide medicament, and a bacteria tight coronal restoration represent a viable technique for capping the inflamed pulp. PMID- 12542101 TI - Safety issues when using a 16% carbamide peroxide whitening solution. AB - BACKGROUND: The scientific literature is lacking on the occurrence of side effects and other safety issues when using carbamide peroxide whitening solutions of concentrations greater than 10%. This double-blind nightguard vital bleaching study compares safety issues when using 16% carbamide peroxide against a placebo or 10% carbamide peroxide (Nite White Classic by Discus Dental Inc.). Evaluated were changes in gingival index, plaque index, nonmarginal gingival index, nongingival oral mucosal index, tooth vitality, and the patients' perceptions of tooth sensitivity and gingival irritation. MATERIALS AND METHODS: Twenty female dental hygiene students participated in the study. Each participant wore a maxillary treatment tray for 1 week without any solution and then for 8 to 10 hours per night for 14 nights, filling each quadrant with placebo, 10% carbamide peroxide, or 16% carbamide peroxide, using a split tray design. RESULTS: With respect to gingival index, plaque index, nonmarginal gingival index, nongingival oral mucosa index, tooth vitality, and tooth sensitivity, there were no statistically significant differences between the 16% carbamide peroxide solution and the other two solutions (p > .05). Quadrants receiving the 16% carbamide peroxide solution experienced more gingival irritation than quadrants receiving placebo or 10% carbamide peroxide solution (p > .05). CONCLUSIONS: When evaluating the above-mentioned safety issues, except for gingival irritation, there were no statistically significant differences between a 16% carbamide peroxide solution and 10% carbamide peroxide solution or a placebo when used as described here. CLINICAL SIGNIFICANCE: Among the 20 participants whose data were analyzed, it was found that a 16% carbamide peroxide whitening solution (Nite White Classic), when used as described in this study, can be effective in nightguard vital bleaching with no statistical differences in gingival index, plaque index, nonmarginal gingival index, nongingival oral mucosa changes, tooth vitality, or tooth sensitivity, compared with a 10% whitening solution (Nite White Classic). More gingival irritation was experienced with 16% carbamide peroxide. Additionally, 20% of the participants in this study self-reported sensitivity when wearing their treatment tray without any solution, and 36% of the participants reported sensitivity to the placebo solution. PMID- 12542103 TI - Longevity of ceramic inlays/onlays, Part I. PMID- 12542102 TI - In vitro wear simulation measurements of composite versus resin-modified glass ionomer luting cements for all-ceramic restorations. AB - BACKGROUND: Although composite cements are generally indicated for cementation of all-ceramic restorations, some manufacturers of dental cements propose that resin modified glass ionomers (RMGIs) may be used for cementation of high-strength ceramic restorations. PURPOSE: This study was undertaken to compare the in vitro abrasion and attrition wear of two dual-cure cements (in dual-cure and self-cure modes) and two RMGI cements when placed between ceramic and enamel to simulate the margin of a restoration. METHODS: Rectangular fragments of pressed ceramic (Empress 2) were cemented between the halves of bovine incisors sectioned mesiodistally, using one of the following materials: RelyX ARC, Variolink II, RelyX Luting, or ProTec CEM. The two resin cements were tested in dual-cure and self-cure modes. A three-body wear test was performed in the new Oregon Health Sciences University (OHSU) oral wear simulator (100,000 cycles; abrasion load: 20 N; attrition load: 90 N). Degree of conversion of resin cements was determined by Fourier transform infrared spectroscopy. Results were analyzed by analysis of variance and Tukey's test (p = .05). Epoxy replicas of wear specimens were observed in the scanning electron microscope. RESULTS: No significant differences in abrasion wear (RelyX ARC dual-cure: 11 +/- 4.4 microm; RelyX ARC self-cure: 17 +/- 7.0 microm; Variolink dual-cure: 14 +/- 8.6 microm; Variolink self-cure: 23 +/- 10.7 microm) or attrition wear (RelyX ARC dual-cure: 18 +/- 6.4 microm; RelyX ARC self-cure: 31 +/- 4.5 microm; Variolink dual-cure: 32 +/- 6.8 microm; Variolink self-cure: 39 +/- 15.9 microm) were found between activation modes of the resin cements. ProTec CEM (32 +/- 8.7 microm) showed abrasion similar to that of Variolink II and RelyX ARC self-cure. Resin-modified glass ionomers showed more attrition wear than the resin cements (ProTec CEM: 62 +/- 13.0 microm; RelyX Luting: 69 +/- 7.1 microm). RelyX ARC showed a similar degree of conversion for both activation modes (dual-cure: 70 +/- 4.3%; self-cure: 68 +/- 1.2%), but Variolink II had a higher degree of conversion in dual-cure mode (67 +/- 0.5% vs 60 +/- 1.0%). Cement wear was accompanied by marginal breakdown and increased surface roughness of enamel and ceramic. CONCLUSIONS: The activation mode of resin cements did not influence their wear resistance. The RMGIs underwent higher attrition wear than the resin cements. CLINICAL SIGNIFICANCE: Increased submargination associated with marginal breakdown and increased roughness of the surrounding structures may be expected when ceramic inlays are cemented with resin-modified glass ionomers. PMID- 12542104 TI - Noncarious cervical defects. PMID- 12542105 TI - Suicidology--expanding its research tools. PMID- 12542106 TI - Suicide, gender, and age variations in India. Are women in indian society protected from suicide? AB - A new set of data concerning the pattern of suicide in India between 1991-1997 are presented. Suicide rates rose over this period despite a small decline in the Indian suicide rate in 1995 and 1996. It was found that between 1995 and 1997 there was a modest fall in the suicide rates among under 29-year-olds of both sexes, and an increase among those 30 years and older. The pattern of suicides in India is bimodal: the incidence of suicides is highest for those in the 30-44 year-old category of both sexes and tends to decline in higher age categories. Suicide rates were nearly equal for young women and men, a contrast with the pattern of suicide sex ratios in eight developed countries. PMID- 12542107 TI - Suicide on the Indian subcontinent. AB - The Indian subcontinent comprises eight countries (India, Pakistan, Bangladesh, Nepal, Sri Lanka, Afghanistan, Bhutan, and the Maldives) and a collective population of more than 1.3 billion people. 10% of the world's suicides (more than 100,000 people) take place in just three of these countries, viz. India, Sri Lanka, and Pakistan. There is very little information on suicides from the other four countries. Some differences from suicides in Western countries include the high use of organophosphate insecticides, larger numbers of married women, fewer elderly subjects, and interpersonal relationship problems and life events as important causative factors. There is need for more and better information regarding suicide in the countries of the Indian subcontinent. In particular, studies must address culture-specific risk factors associated with suicide in these countries. The prevention of this important public health problem in an area of the world with myriad socio-economic problems, meager resources, and stigmatization of mental illness poses a formidable challenge to mental health professionals, policy makers, and governments of these countries. PMID- 12542108 TI - Suicidal behavior in South Asians in the UK. AB - Rates of deliberate self-harm have been reported to be higher among South Asians, especially women, in the UK. Two studies collected information on inception rates of deliberate self-harm in the UK and reported that rates of attempted suicide are much higher among South Asian women especially those aged 18-24. These rates are associated with high rates of cultural alienation and previous attempts. The implications of cultural conflict are discussed in this context. In addition, a hypothetical model of interrelationship of factors for both adults and adolescents is described. PMID- 12542109 TI - Beyond risk theory: suicidal behavior in its social and epidemiological context. AB - The effect of exposure to risk factors for suicidal behavior varies from place to place and from period to period. This may be due to contextual influences, which arise if individuals' suicide risk depends not only on their personal exposure to risk or protective factors, but also on how these are distributed in their social, cultural, economic, or even physical environments. There has been relatively little explicit attention in suicide research for such contextual influences even though they are an important component of the cross-level bias, which can arise when aggregate level associations are assumed to also apply in individuals and vice versa. Contextual effects are conceptually related to the issues of social selection vs. causation, population density, and network effects. Because of a lack of prospective multilevel research, it is unclear exactly which mechanisms underlie the phenomenon that the distribution of risk factors in an individual's context may affect their suicide risk above and beyond their personal exposure. A number of mechanisms, like deviancy amplification, formalization of restraints, and buffering effects of social support are proposed. Contextual effects may result in a concentration of suicide risk in persons when the risk factors they are exposed to become rare--whether spontaneously or through focused prevention. This has important but mostly overlooked implications for population-based prevention strategies. PMID- 12542110 TI - Evidence-based prevention strategies of suicide behavior in the world: implications for Slovenia. AB - In this review we consider how Slovenia could consider tackling its high rate of suicide (overall 29 per 100,000, 46 in males, 13 in females). First, we consider the evidence for risk factors that may contribute to Slovenia's high rate of suicide. Second, we describe the interventions to try to reduce the impact of these factors and the evidence for such interventions. We categorize interventions in terms of their operation at either the population level or that of high-risk groups. However, it should be borne in mind that settings often assumed to provide access to population groups, such as general practice and schools, do not reach some people who are likely to be at high risk; for example those who have dropped out of school or who have been excluded from a GP's list. We focus particularly on those for high-risk groups, as a number of East-European countries with high suicide rates such as Slovenia, Hungary, and the Baltic republics are currently considering a shift toward more community-based mental health services. The provision of community mental health services in Slovenia would provide an opportunity to study their impact on the suicide rate. However, we conclude that their development should be accompanied by other initiatives operating at population levels. This multilevel approach acknowledges the complexity of the etiology of suicide, the impossibility of reaching all those at risk through services and the lack of strong evidence for any one intervention. PMID- 12542111 TI - Depression and parasuicide in refugee and Slovenian adolescents. AB - The association between depression and traumatic experiences, stress, loss, and bereavement is well known. It was expected that Bosnian refugee adolescents who had fled from war zones to Slovenia would develop higher levels of depression than their Slovenian peers without war traumatic experiences. Two years after the beginning of the war in Bosnia, 265 8th-grade refugee students aged 14 to 15 years were assessed with the Children's Depression Inventory (CDI) and compared with a sample of 195 Slovenian students of the same age. The Bosnian refugees showed significantly lower rates of depressive symptomatology despite very high rates of posttraumatic stress symptoms. Slovenian adolescents showed significantly lower self-esteem and greater feelings of inadequacy in the academic field; they were more likely to have feelings of not being loved and more often expressed the wish to kill themselves. Refugees described more sadness, worried about the future and about physical pains, but they do not show lower school achievement or more behavioral problems. The results indicate that the association between traumatic experiences and depressive symptoms is not inevitable, and that other factors including culture and context influences must be taken into account. PMID- 12542112 TI - The scientific study of suicide requires accurate data. PMID- 12542113 TI - Progress--what progress? Letter across the Pacific. PMID- 12542114 TI - Comment on Burless and DeLeo. PMID- 12542115 TI - Evolution of the mortality rate in the age class of 75-84 years worldwide. PMID- 12542116 TI - Right ventricular function in patients with acute anterior myocardial infarction: tissue Doppler echocardiographic approach. AB - OBJECTIVE: Our purpose was to investigate the right ventricular (RV) performance of patients with a first acute anterior myocardial infarction (AAMI) by using pulsed wave Doppler tissue (PWDT) samplings of tricuspid annulus and RV free wall. METHODS AND RESULTS: The study group included 31 patients with AAMI and 20 age-matched controls. Conventional indexes of RV functions were the magnitude of tricuspid annular plane systolic excursion (TAPSE), and the transpulmonary and transtricuspid Doppler parameters. PWDT velocities were obtained by placing the sample volume at the lateral tricuspid annulus and the mid-segment of RV free wall; the peak systolic (S), early (E) and late (A) diastolic PWDT velocities and time intervals from ECG-Q wave to their peaks were analysed. Standard indexes were comparable except TAPSE that was significantly lower in AAMI-patients (p < 0.001). S velocities were similar; A of both regions (p = 0.018 and 0.012) and E of RV free wall (p = 0.011) were significantly increased in AAMI-group. Q-Sa intervals in both regions (p = 0.007 and 0.015) and Q-Ea of tricuspid annulus (p = 0.045) were significantly shorter in patients with AAMI. TAPSE and E of RV free wall had significant negative correlations with left ventricular systolic volume index and right atrial filling fraction (AFF), respectively (r = -0.46, p = 0.01 for both). A of tricuspid annulus had a positive correlation with left AFF (r = 0.42, p = 0.02). CONCLUSION: PWDT imaging of tricuspid annulus and RV free wall is capable to sensitively detect the adaptive mechanisms and unfavourable diastolic properties of RV dynamics in patients with AAMI. PMID- 12542117 TI - Maternal reproductive history and isolated hypoplastic left heart syndrome. PMID- 12542118 TI - Lack of association between plasma lipoprotein(a) concentrations and the presence or absence of coronary atherosclerosis. AB - BACKGROUND: Findings from previous studies relating lipoprotein(a) [Lp(a)] as an independent risk factor for coronary atherosclerosis and the presence of angiographically detectable coronary atherosclerotic lesions are not consistent. This study was performed to determine whether the plasma concentration of Lp(a) is associated with coronary atherosclerosis asessed by coronary angiography. METHODS: We studied a total of 100 men and women (41 women, 59 men, age 63.7 +/- 11.0 years) who were referred for coronary angiography. Base-line data collection comprised conventional risk factors for coronary artery disease, lipids, fasting total homocysteine, and clinical characteristics. The relation between plasma Lp(a) levels and the presence or absence of coronary lesions was studied. The coronary angiograms were evaluated in a blinded manner. Any coronary stenosis was considered as coronary artery disease (CAD). RESULTS: From the 100 patients, 40 were found to have no CAD and 60 had CAD assessed by coronary angiography. Estimates of the relative risk of coronary heart disease for the fifth quintile of plasma Lp(a) as compared with the first quintile were 0.87 (95 percent confidence interval, 0.66 to 1.34). After adjustment for age, sex, lipoproteins, and homocysteine levels, estimates of the relative risk of coronary heart disease for the fifth quintile of plasma Lp(a) as compared with the first quintile were 1.06 (95 percent confidence interval, 0.81 to 1.39). The presence of angiographic CAD was associated with patient age (p=0.048), male sex (p<0.01), high LDL cholesterol levels (p=0.02), low HDL-cholesterol levels (p=0.02), high plasma fibrinogen levels (p<0.01) and high fasting total homocysteine levels (p=0.04). CONCLUSION: These results suggest that the plasma concentration of Lp(a) is not associated with the presence of coronary artery disease in patients referred for coronary angiography. PMID- 12542119 TI - Hyperhomocysteinaemia and coronary artery disease in the Turkish population. AB - OBJECTIVE: Many studies have demonstrated a strong association between elevated plasma total homocysteine (Hcy) levels and vascular disease. The objective of this study was to examine the relation between homocysteine levels and coronary artery disease in Turkish patients. METHODS AND RESULTS: In this study plasma homocysteine levels were measured in control and patient groups. A significant coronary artery lesion was defined as a stenosis of > or = 70% as shown by coronary angiography and determined by on-line quantitative measurements; treatment was by coronary angioplasty. Total plasma Hcy level was measured before the coronary intervention. Plasma homocysteine levels were measured by an HPLC method in patients with a definite diagnosis of coronary artery disease and compared with age- and sex-matched controls. Patients with coronary artery disease had significantly higher mean homocysteine concentrations than control subjects (geometric mean +/- 95% CI: 12.5 +/- 1.1 micromol/l vs. 8.60 +/- 1.07 micromol/l, p<0.001). Eighty-three (59%) members of the patient group and 14 (21%) members of the control group had plasma homocysteine concentrations above the 11.3 micromol/l, which represents the concentration which includes the uppermost quintile of the control group distribution (odds ratio 4.35, 95% CI; 2.1-8.94). CONCLUSION: Results of this study indicate that high plasma levels of homocysteine in Turkish subjects are associated with coronary artery disease. Our data suggest that focusing public health initiatives on this issue may reduce the high prevalence of cardiovascular disease in the Turkish population. PMID- 12542120 TI - Plasma fibronectin level and its relationships with lipids, lipoproteins and C reactive protein in patients with dyslipidaemia during lipid-lowering therapy. AB - OBJECTIVES: The purpose of this study is to determine plasma fibronectin level and its relationships with plasma lipids, lipoproteins and C-reactive protein (CRP) levels in patients with dyslipidaemia during lipid-lowering therapy. METHODS: Plasma levels of fibronectin, CRP, fibrinogen, lipids and lipoproteins in 38 patients with dyslipidaemia were determined before and after lipid-lowering therapy by using atorvastatin, 10 mg/day. RESULTS: After lipid-lowering therapy, serum levels of fibronectin and CRP were found to be significantly decreased by 30.4% and 43.6%, respectively, while fibrinogen levels were increased 11.7% in patients with dyslipidaemia. Before the treatment, fibronectin was found to be positively correlated with CRP and total cholesterol (r=0.38, p<0.05 and r=0.33, p<0.05, respectively) and negatively correlated with high-density lipoprotein cholesterol (HDL-C) (r=-0.42, p<0.01) in patients with dyslipidaemia. High fibronectin levels (0.57 +/- 0.17 g/l) were found in patients with HDL-C below 35 mg/dl (0.57 +/- 0.09 g/l), compared to patients with HDL-C above 35 mg/dl (0.45 +/- 0.11 g/l). During the lipid-lowering therapy, total cholesterol, low-density lipoprotein cholesterol (LDL-C), triglyceride and apo B levels were reduced while HDL-C and apo AI levels were increased. CONCLUSIONS: It was found that plasma fibronectin and CRP levels were decreased by lipid lowering therapy. Plasma fibronectin levels were associated with lipids, lipoproteins, CRP levels before treatment and these relationships disappeared after treatment. Consequently, it was suggested that reduction of plasma fibronectin levels, together with lipids and loss of its relationship with CRP, may play a role on the antiatherogenic effects of lipid-lowering therapy. PMID- 12542121 TI - Interhospital transport of haemodynamically unstable patients using intra-aortic balloon pumping. AB - The intra-aortic balloon pump (IABP) is a valuable tool in the treatment of cardiogenic shock or instable myocardial ischaemia. We report on our recent experience with interhospital transport of haemodynamically unstable patients using on-site stabilization with IABP. A 75% survival rate encourages us to keep using this technique despite the logistic efforts it demands. PMID- 12542122 TI - Accessory mitral valve tissue as a cause of left ventricular outflow tract obstruction and probable cause of cerebrovascular accident. AB - Accessory mitral valve tissue is a rare finding in echocardiographic examinations. We report a case with an echocardiographically detected accessory mitral valve tissue which caused left ventricular outflow tract obstruction and was presumably responsible for a cerebrovascular accident. PMID- 12542123 TI - Cardiac pseudoaneurysm coupled with a rupture of the papillary muscle complicating myocardial infarction. AB - As a complication of myocardial infarction, dual rupture of the left ventricular myocardium and the papillary muscle is a rare condition. In such a case, the heart is predisposed to reduced output because of unloading of the ventricle during systole, resulting in the patient being in danger of deteriorating into a severe state or dying suddenly from cardiogenic shock. We report a rescued case of a 65-year-old woman, who had cardiogenic shock due to left ventricular pseudoaneurysm, coupled with partial rupture of the posteromedial papillary muscle three weeks after posterior myocardial infarction. Emergent left ventriculography revealed a large aneurysmal cavity and regurgitation towards the left atrium. The patient underwent emergent aneurysmectomy with mitral valve replacement. PMID- 12542124 TI - Acute myocardial infarction secondary to thyrotoxicosis. AB - Angina and myocardial infarction have been reported in hyperthyroidism with normal coronary arteries, presumably secondary to thyroid-induced coronary artery spasm. We report the case of a very young female patient with an atypical presentation of acute myocardial infarction mimicking pulmonary embolization secondary to thyrotoxicosis. Hyperthyroidism should be kept in mind in younger patients in the differential diagnosis of coronary artery disease. PMID- 12542125 TI - Arrhythmogenic right ventricular cardiomyopathy and atrial right-to-left shunt. AB - A 47-year-old woman diagnosed with arrhythmogenic right ventricular cardiomyopathy, was studied at our institution with palpitations, progressive dyspnoea and cyanosis over a 3-year period. Her haemoglobin saturation was 76.1%. The transthoracic echocardiogram showed that both right atrium and ventricle were dilated, with diffuse akinetic and hypokinetic areas. Radionuclide scintigraphy, angiography and magnetic resonance imaging showed dilated and severe dysfunction of the right ventricle, with depressed ejection fraction (0.16), and bulging at the level of the right ventricular anterior wall areas. Doppler examination revealed a right-to-left interatrial shunt through a patent foramen ovale. Contrast transoesophageal echocardiography confirmed the right-to-left shunting. Considering the clinical symptoms were severe and refractory to medical treatment, she was referred for cardiac transplantation. Unfortunately the patient died suddenly a few weeks later. PMID- 12542126 TI - A massive left-to-right shunt due to a ruptured giant aneurysm of the sinus of Valsalva. AB - We describe a patient with a giant ruptured sinus of Valsalva aneurysm causing a massive left-to-right shunt. The diagnosis was made by transoesophageal echocardiography and confirmed by angiography. We outline briefly a diagnostic and therapeutic work-up. PMID- 12542127 TI - Blinks of the mind: memory effects of attentional processes. AB - If 2 words are presented successively within 500 ms, subjects often miss the 2nd word. This attentional blink reflects a limited capacity to attend to incoming information. Memory effects were studied for words that fell within an attentional blink. Unrelated words were presented in a modified rapid serial visual presentation task at varying stimulus-onset asynchronies, and attention was systematically manipulated. Subsequently, recognition, repetition priming, and semantic priming were measured separately in 3 experiments. Unidentified words showed no recognition and no repetition priming. However, blinked (i.e., unidentified) words did produce semantic priming in related words. When, for instance, ring was blinked, it was easier to subsequently identify wedding than apple. In contrast, when the blinked word itself was presented again, it was not easier to identify than an unrelated word. Possible interpretations of this paradoxical finding are discussed. PMID- 12542128 TI - The octave illusion revisited: suppression or fusion between ears? AB - The octave illusion occurs when each ear receives a sequence of tones alternating by 1 octave but with the high and low tones in different ears. Most listeners perceive these stimuli as a high pitch in one ear alternating with a low pitch in the other ear. D. Deutsch and P. L. Roll (1976) interpreted this phenomenon as evidence for a what-where division of auditory processing caused by sequential interactions between the tones. They argued that the pitch follows the frequency presented to the dominant ear but is lateralized toward the higher frequency component. This model was examined in 4 experiments. Results indicate that the perceived pitch approximates the fundamental frequency and that the illusion does not depend on sequential interactions. The octave illusion may arise from an interaction between dichotic fusion and binaural diplacusis rather than from suppression as proposed by Deutsch. PMID- 12542129 TI - Visual search for dimensionally redundant pop-out targets: parallel-coactive processing of dimensions is location specific. AB - Three visual search experiments investigated redundancy gains for single and dual odd-one-out feature targets that differed from distractors in orientation, color, or both. In Experiment 1, redundant-target displays contained (a) a single target defined in 2 dimensions, (b) dual targets each defined in a different dimension, or (c) dual targets both defined in the same dimension. The redundancy gains, relative to single nonredundant targets, decreased from the first condition on, with violations of J. Miller's (1982) race model inequality (RMI) manifested only in the first 2 conditions. Experiment 2 systematically varied the spatial separation between dual targets each defined in a different dimension. Violations of the RMI were evident only when the 2 targets occupied nearby locations. Experiment 3 provided evidence of RMI violations by dimensionally redundant targets at both precued (likely) and noncued (unlikely) display locations. Taken together, these results suggest that there is coactivation of a common mechanism by target signals in different dimensions (not by signals in the same dimension), that the coactivation effects are spatially specific, and that the coactivated mechanisms are located at a preattentive, perceptual stage of processing. PMID- 12542130 TI - Bigger than a breadbox? Attention to distractors may not enhance negative priming. AB - In several recent studies, P. A. MacDonald and colleagues (e.g., P. A. MacDonald & S. Joordens, 2000) reported unusually large negative priming effects and claimed that attention to distractors, counter to expectations, served to enhance the magnitude of the effect. In 3 experiments using their novel comparative judgment task, negative priming was assessed using both a control condition based on that of P. A. MacDonald, S. Joordens, and K. N. Seergobin (1999) and one in which control probe items exactly matched those on ignored repetition trials. In MacDonald et al.'s unmatched control condition, participants were faster than on ignored repetition trials, but this difference was reduced or absent when control items were matched. This result led to the conclusion that the apparently large negative priming effect reported by MacDonald and colleagues may be an artifact arising because judgments for a subset of their control trials were relatively easier than for ignored repetition trials. PMID- 12542131 TI - Feature-specific perceptual processing dissociates action from recognition. AB - Does visual processing differ for action and recognition? To address this question, the authors capitalized on research showing that color is preferred over binocular disparity in the ventral (recognition) stream, whereas disparity is preferred over color in the dorsal (action) stream. Participants searched for oblique targets among vertical distractors in displays defined only by color or disparity. Action-task participants stamped the target with a handheld block, whereas recognition-task participants lifted the block through a target compatible gap. Analyses of reaction time and time-varying hand orientation showed that disparity and color displays were processed equally efficiently during action, but disparity was processed less efficiently than color during recognition. The results suggest that visual processing differs for action and recognition. PMID- 12542132 TI - Control over location-based response activation in the Simon task: behavioral and electrophysiological evidence. AB - In 4 Simon experiments the authors examined control over 2 routes of sensorimotor processing: response priming in the unconditional route and response selection via the conditional route. The Simon effect diminished as the frequency of noncorresponding trials increased. Location-based response priming was observed only when the stimulus followed a corresponding event but not after a noncorresponding trial. Therefore, the unconditional route appears to be suppressed whenever the task context indicates priming as potentially disadvantageous. Moreover, the task-irrelevant stimulus location was used for response selection as a function of correspondence probability. Although exact repetitions of stimulus-response sequences caused a marked speed-up of responses, this 3rd mechanism is independent of unconditional route suppression and frequency-based adjustments in the conditional route. PMID- 12542133 TI - Online response preparation in a rapid serial visual search task. AB - Online response preparation was assessed in a visual search task using rapid serial visual presentation. In each trial, a series of letters was presented sequentially, and participants were instructed to make a target-present response if a prespecified target letter was presented or a target-absent response if it was not. Measurements of response preparation using both probe reaction time and the lateralized readiness potential indicated that preparation of the target absent response increased near the end of the sequence. Most of the increase appeared to be due to direct priming of the target-absent response by nontarget letters, but part was due to the increased conditional probability of this response near the end of the sequence. These results extend previous studies of response preparation by showing online response preparation during a temporally extended reaction time task. PMID- 12542134 TI - Neural correlates of spatial term use. AB - The current research used event-related brain potentials (ERPs) to assess the processes underlying online apprehension of the spatial term above. Constituent steps defined within G. D. Logan and D. D. Sadler's (1996) computational theory of apprehension were associated with distinct modulations of ERPs. Specifically, finding the relevant objects was associated with an amplitude modulation of P3; competition in assigning directions to space was associated with modulation of a frontal slow wave; and computing and comparing the spatial relation was associated with modulation of a parietal slow wave. These modulations were differentially influenced by the type of reference frame used to define the spatial term and by the participant's response. The current study supports this decompositional approach to apprehension and provides a means of assessing each step independently. PMID- 12542135 TI - The role of orthographic and phonological codes in the word and the pseudoword superiority effect: an analysis by means of multinomial processing tree models. AB - Central to the current accounts of the word and the pseudoword superiority effect (WSE and PWSE, respectively) is the concept of a unitized code that is less susceptible to masking than single-letter codes. Current explanations of the WSE and PWSE assume that this unitized code is orthographic, explaining these phenomena by the assumption of dual read-out from unitized and single-letter codes. In this article, orthographic dual read-out models are compared with a phonological dual read-out model (which is based on the assumption that the 1st unitized code is phonological). From this phonological code, an orthographic code is derived, through either lexical access or assembly. Comparison of the orthographic and phonological dual read-out models was performed by formulating both models as multinomial processing tree models. From an application of these models to the data of 2 letter identification experiments, it was clear that the orthographic dual read-out models are insufficient as an explanation of the PWSE, whereas the phonological dual read-out model is sufficient. PMID- 12542136 TI - Global effect of a nearby distractor on targeting eye and hand movements. AB - Eye-hand coordination was investigated with the global effect paradigm. In this paradigm, saccades typically land in between the target and a nearby presented distractor, the configuration's center of gravity. This so-called global effect, or spatial averaging, is attributed to incomplete target selection. Four experiments demonstrated a similar effect for hand movements; thus, eye and hand are coupled during target selection. However, under some conditions the global effect was different for eye and hand, suggesting that their coupling is not achieved through a shared target representation. Instead, eye and hand seem to use 2 separate target representations that exchange information. The convergent amplitudes of eye and hand with simultaneous execution support this interpretation. Latencies showed a similar converging pattern. PMID- 12542138 TI - Monocular motion adaptation affects the perceived trajectory of stereomotion. AB - Perceived stereomotion trajectory was measured before and after adaptation to lateral motion in the dominant or nondominant eye to assess the relative contributions of 2 cues: changing disparity and interocular velocity difference. Perceived speed for monocular lateral motion and perceived binocular visual direction (BVD) was also assessed. Unlike stereomotion trajectory perception, the BVD of static targets showed an ocular dominance bias, even without adaptation. Adaptation caused equivalent biases in perceived trajectory and monocular motion speed, without significantly affecting perceived BVD. Predictions from monocular motion data closely match trajectory perception data, unlike those from BVD sources. The results suggest that the interocular velocity differences make a significant contribution to stereomotion trajectory perception. PMID- 12542137 TI - Learning to recognize talkers from natural, sinewave, and reversed speech samples. AB - In 5 experiments, the authors investigated how listeners learn to recognize unfamiliar talkers and how experience with specific utterances generalizes to novel instances. Listeners were trained over several days to identify 10 talkers from natural, sinewave, or reversed speech sentences. The sinewave signals preserved phonetic and some suprasegmental properties while eliminating natural vocal quality. In contrast, the reversed speech signals preserved vocal quality while distorting temporally based phonetic properties. The training results indicate that listeners learned to identify talkers even from acoustic signals lacking natural vocal quality. Generalization performance varied across the different signals and depended on the salience of phonetic information. The results suggest similarities in the phonetic attributes underlying talker recognition and phonetic perception. PMID- 12542139 TI - Within- and between-dimensional processing in the auditory modality. AB - Participants made speeded target-nontarget responses to singly presented auditory stimuli in 2 tasks. In within-dimension conditions, participants listened for either of 2 target features taken from the same dimension; in between-dimensions conditions, the target features were taken from different dimensions. Judgments were based on the presence or absence of either target feature. Speech sounds, defined relative to sound identity and locale, were used in Experiment 1, whereas tones, comprising pitch and locale components, were used in Experiments 2 and 3. In all cases, participants performed better when the target features were taken from the same dimension than when they were taken from different dimensions. Data suggest that the auditory and visual systems exhibit the same higher level processing constraints. PMID- 12542140 TI - The optic trajectory is not a lot of use if you want to catch the ball. AB - According to linear optic trajectory (LOT) theory, fielders use the direction of curvature of theoptic trajectory to control the way they run to intercept the ball. Data presented by D. M. Shaffer and M. K. McBeath (2002) as support for LOT theory show that the optic trajectory of balls that will fall behind the fielder provide the cue that LOT theory predicts would send the fielder running forward, not backward. In this article, the authors show that watching these balls would provide the fielder with the cue that the optic acceleration cancellation (OAC) theory of interception predicts would send the fielder running backward. It appears that the fielders studied by Shaffer and McBeath were following the cue predicted by OAC theory, not that predicted by LOT theory. PMID- 12542141 TI - Cow-calf and feedlot performances of Brahman-derivative breeds. AB - A study was conducted to compare Brangus, Beefmaster, Gelbray, and Simbrah breed influences for economically important traits. Brangus (9), Beefmaster (12), Gelbray (10), and Simbrah (7) sires were used in purebred and crossbred (Brahman x Hereford F1 cows) matings to generate calves (326) in eight breed groups. Beefmaster cows were of similar size (448 kg), Brangus and Gelbray cows were 11% heavier (501 and 503 kg), and Simbrah cows were 21% heavier (548 kg) compared to Brahman x Hereford F1 cows (452 kg). Calves sired by Brangus and Beefmaster bulls had lower birth weights (35 vs 38 kg; P < 0.05), preweaning growth rates (0.87 vs 0.91 kg x d(-1); P < 0.01), and weaning weights (206 vs 219 kg; P < 0.01) than Gelbray- and Simbrah-sired calves. Birth weights, preweaning ADG, and weaning weight and hip heights were similar between Brangus- and Beefmaster-sired calves. Simbrah-sired calves had greater preweaning growth rates (0.94 vs 0.88 kg x d( 1); P < 0.05), weaning weights (227 vs 211 kg; P < 0.01), and adjusted 205-d hip heights (126 vs 122 cm; P < 0.05) than Gelbray-sired calves. Straightbred Angus steers were introduced in the postweaning portion of the study. Steer calves were placed on feed at an average age of 14.5 mo. Steers were removed from the feedlot upon attaining a targeted 10 mm of backfat. Feedlot ADG did not differ among sire breeds. Brahman-derivative sired steers required an additional 54 d on feed (P < 0.01) and were 86 kg heavier (P < 0.01) at harvest than Angus steers. Continental Brahman steers spent an additional 25 d on feed (P < 0.05) and were 35 kg heavier (P < 0.01) at harvest than British-Brahman steers. Simbrah-sired steers were 52 kg heavier (P < 0.01) at harvest than Gelbray-sired steers when fed for a similar number of days (211 vs 203 d). However, straightbred Simbrah steers required an additional 12 d on feed (P < 0.01) and weighed 47 kg more (P < 0.01) than Simbrah sired crossbred steers. The economic value of the heavier calf weaning weights may be offset by the attendant larger cow size of the Continental-Brahman compared to the British-Brahman breeds. Similarly, the heavier weights of Continental-Brahman compared to British-Brahman steers, when harvested at a prescribed level of fatness may be viewed as a benefit, but the increased number of requisite days in the feedlot is a disadvantage. PMID- 12542142 TI - Prediction of intramuscular fat percentage in live swine using real-time ultrasound. AB - Purebred Durocs (n = 207) were used to develop a model to predict loin intramuscular fat percentage (PIMF) of the longissimus muscle in live pigs. A minimum of four longitudinal, real-time ultrasound images were collected 7 cm off midline across the 10th to the 13th ribs on the live animal. A trained technician used texture analysis software to interpret the images and produce 10 image parameters. Backfat and loin muscle area were measured from a cross-sectional image at the 10th rib. After harvest, a slice from the 10th to the 11h rib loin interface was used to determine carcass loin intramuscular fat percentage (CIMF). The model to predict loin intramuscular fat percentage was developed using linear regression analysis with CIMF as the dependent variable. Initial independent variables were off-test weight, live animal ultrasonic 10th rib backfat and loin muscle area, and the 10 image parameters. Independent variables were removed individually until all variables remaining were significant (P < 0.05). The final prediction model included live animal ultrasound backfat and five image parameters. The multiple coefficient of determination and root mean square error for the prediction model were 0.32 and 1.02%, respectively. An independent data set of Duroc (n = 331) and Yorkshire (n = 288) pigs from two replications of the National Pork Board's Genetics of Lean Efficiency Project were used for model validation. Results showed the Duroc pigs provided the beat validation of the model. The product moment correlation and rank correlation coefficients between PIMF and CIMF were 0.60 and 0.56, respectively, in the Duroc population. Results show real-time ultrasound image analysis can be used to predict intramuscular fat percentage in live swine. PMID- 12542143 TI - Seasonal effects on estrous behavior and time of ovulation in nonlactating beef cows. AB - Estrous behavior and time of ovulation relative to the onset of estrus were determined in mature Angus x Hereford cows (n = 17 to 21 each season) during summer, winter, and spring for 2 yr. Estrous behavior was evaluated during the first of two consecutive estrous periods, and time of ovulation was determined during the second estrus. Concentrations of progesterone were quantified in twice weekly blood samples to ensure all cows had normal estrous cycles. The HeatWatch system was used to measure the duration of estrus, number of mounts received per estrus, and duration of the longest interval between mounts received. Commencing 16 h after the onset of the second estrus, transrectal ultrasonography was performed every 4 h until the dominant follicle was no longer present on the ovary, and time of ovulation was defined as 2 h preceding the absence of the dominant follicle. There was a seasonal effect on the duration of estrus; cows were estrus longer in summer (17.6 +/- 0.8 h) than in winter (15.5 +/- 0.8 h; P = 0.07) or spring (13.9 +/- 0.9 h; P < 0.05). Cows were mounted more times per estrus (P < 0.05) in winter (59.0 +/- 5.3) than in summer (43.6 +/- 5.3) or spring (38.2 +/- 5.8). Intervals between mounts of estrous cows were longer (P < 0.05) in summer (4.1 +/- 0.4 h) than in spring or winter (2.7 +/- 0.4 h). During all seasons, cows were mounted more times (P < 0.01) between 0600 to 1200 (3.2 +/ 0.2 mounts received/h of estrus) than during other times of the day (2.1 +/- 0.2 mounts received/h of estrus). Cows ovulated 31.1 +/- 0.6 h after the onset of estrus, and time of ovulation was not influenced by season. We conclude that season influences estrous behavior of beef cows; cows are mounted more times per estrus in winter than in summer or spring. Time of ovulation relative to the onset of estrus is constant during all seasons and averages 31.1 h. PMID- 12542144 TI - Evaluation of a fixed-time artificial insemination protocol for postpartum suckled beef cows. AB - Treatment with melengestrol acetate (MGA), an oral progestin, prior to administration of gonadotropin-releasing hormone (GnRH) and prostaglandin F2alpha (PG) effectively synchronizes estrus and maintains high fertility in postpartum beef cows. The objective of this experiment was to determine whether treatment with MGA prior to a GnRH-PG-GnRH protocol would improve pregnancy rates resulting from fixed-time artificial insemination (AI). Multiparous crossbred beef cows at two University of Missouri-Columbia farms (n = 90 and n = 137) were assigned by age and days postpartum to one of two treatments. Cows were fed carrier (1.8 kg x animal(-1) x d(-1)) with or without MGA (0.278 mg x kg(-1)) for 14 d. All cows were administered GnRH (100 microg; intramuscularly) on d 12 after MGA or carrier withdrawal and 7 d before PG (25 mg; intramuscularly). All cows received a second injection of GnRH and AI 72 h after PG. Mean days postpartum for MGA and control cows at the initiation of treatment were 39.6 and 38.9 d for herd 1; and 51.9 and 50.9 d for herd 2, respectively (P > 0.70 within herds). Blood samples were collected from all cows at 10 and 1 d before the feeding of MGA or carrier began and at the times GnRH and PG were administered. Concentrations of progesterone in serum at the initiation of treatment were elevated (>1 ng/mL) in 0% of MGA and 7% of control cows in herd 1, and 54% of MGA and 49% of control cows in herd 2 (P > 0.05 within herds). Pregnancy rates to fixed-time AI were determined by transrectal ultrasonography 50 d after AI. Pregnancy rates in herd 1 were 58% (26/45) and 51% (23/45) for MGA-treated and control cows, respectively (P = 0.52), and 63% (44/70) and 45% (30/67) for MGA-treated and control cows in herd 2, respectively (P = 0.03). Differences in pregnancy rates to fixed-time AI were significant (P = 0.04) when data from the two herds were combined (with MGA = 70/115 [61%]; control = 53/112 [47%]). There was no difference (P > 0.20) in final pregnancy rates (timed AI plus 45 d exposure to bulls) between treatments, within herds, or when herds were combined. In summary, pregnancy rates resulting from fixed-time AI may be improved with treatment of MGA prior to a GnRH-PG-GnRH protocol. PMID- 12542145 TI - Effects of providing a water-soluble globulin in drinking water and diet complexity on growth performance of weanling pigs. AB - Two experiments were conducted to evaluate the effects of providing a water soluble globulin in the drinking water on growth performance of weanling pigs. In Exp. 1, 360 weanling pigs (5.0 +/- 1.2 kg; 17 +/- 3 d of age; PIC) were blocked by initial weight and allotted to one of six treatments in a 2 x 3 factorial arrangement. Treatments included three diet complexity regimens with or without water-soluble globulin (3 and 1.5% solutions; d 0 to 7 and d 0 to 14, respectively) provided in the drinking water. The 35-d study was divided into three phases (d 0 to 7, 7 to 14, and 14 to 35) with corresponding lysine levels of 1.6, 1.5, and 1.35%. Soybean meal replaced specialty protein and lactose sources to provide three different complexity regimens. From d 0 to 7, a water soluble globulin x diet complexity interaction (P < 0.05) was observed for average daily gain (ADG) and gain:feed (G/F). Increasing diet complexity increased ADG and G/F for pigs provided water, whereas the medium diet complexity regimen optimized performance for pigs offered water-soluble globulin. From d 0 to 14, pigs fed the two more complex regimens had greater ADG and G/F (P < 0.01) than the pigs fed the least complex regimen. Pigs offered water-soluble globulin had decreased (P < 0.01) ADFI, but increased (P < 0.001) G/F from d 0 to 14. For overall performance (d 0 to 35), increasing diet complexity increased (P < 0.03) ADG and ADFI, whereas water-soluble globulin offered from d 0 to 14 had no effect. In Exp. 2, 360 weanling pigs (5.2 +/- 1.6 kg; 19 +/- 4 d of age) were used in a 21-d growth assay. The trial was arranged as a 2 x 3 factorial with pigs fed the low- or medium-complexity diets (Exp. 1) with water or a 3% solution of water-soluble globulin offered for 4 or 8 d after weaning. From d 0 to 4, pigs offered water-soluble globulin had increased (P < 0.001) ADG and G/F compared with pigs provided water, whereas from d 4 to 8, pigs provided water had increased (P < 0.05) ADG and G/F compared with pigs offered water-soluble globulin. Pigs fed the medium-complexity diet had increased ADG and G/F (d 4 to 8 and d 8 to 12) compared with pigs fed the low-complexity diet. From d 0 to 8 and d 0 to 21, pigs provided water-soluble globulin for 4 or 8 d after weaning had improved G/F compared with pigs provided water. Results demonstrate that providing water-soluble globulin through the water source of weanling pigs improves ADG and G/F immediately after weaning. PMID- 12542146 TI - Genotype x environmental interaction for mature size and rate of maturing for Angus, Brahman, and reciprocal-cross cows grazing bermudagrass or endophyte infected fescue. AB - Mature weight and rate of maturing were estimated in 177 Angus, Brahman, and reciprocal-cross cows grazing bermudagrass or endophyte-infected tall fescue over a 4-yr period to evaluate genotype x environment interactions. Data were collected every 28 d until cows were approximately 18 mo of age and then at prebreeding, postcalving, and weaning of calf. All cows with weight data to at least 42 mo of age were included in the analysis. Mature weight and rate of maturing were estimated using the three-parameter growth curve model described by Brody (1945). Data were pooled over year and analyzed by the general linear model procedure of SAS. Included in the models for mature weight and rate of maturing were the independent variables of genotype, environment, and genotype x environment interaction. There was a genotype x environment interaction (P < 0.01) for mature body weight (BW) but not for rate of maturing. Angus cows grazing fescue pastures had greater (P < 0.01) mean mature BW than Angus x Brahman cows grazing bermudagrass (611 +/- 17 vs 546 +/- 16 kg). Angus x Brahman cows grazing bermudagrass had lower (P < 0.05) mean mature BW than Brahman x Angus cows grazing bermudagrass or endophyte-infected fescue and Brahman cows grazing bermudagrass (546 +/- 16 vs 624 +/- 19, 614 +/- 22 and 598 +/- 20 kg, respectively). Brahman cows grazing endophyte-infected fescue had smaller (P < 0.05) mean mature BW than all genotype x forage combinations except for Angus x Brahman cows grazing bermudagrass. Angus cows had a smaller (P < 0.05) mean rate of maturing than Angus x Brahman and Brahman x Angus cows (0.039 +/- 0.002 vs 0.054 +/- 0.002 and 0.049 +/- 0.002%/mo, respectively), respectively, and Angus x Brahman cows had a larger (P < 0.05) mean rate of maturing than Brahman x Angus and Brahman cows (0.054 +/- 0.002 vs 0.049 +/- 0.002 and 0.041 +/- 0.002 %/mo, respectively). There was a direct breed x forage interaction (P < 0.05) for mature BW. These data suggest that the choice of breed type is important in maintaining a crossbreeding program, in that mature BW and rate of maturing are critical to the matching of animal requirements to available production resources. PMID- 12542147 TI - Evaluation of single-nucleotide polymorphisms in CAPN1 for association with meat tenderness in cattle. AB - Micromolar calcium activated neutral protease (CAPN1) was evaluated as a candidate gene for a quantitative trait locus (QTL) on BTA29 affecting meat tenderness by characterization of nucleotide sequence variation in the gene. Single-nucleotide polymorphisms (SNP) were identified by sequencing all 22 exons and 19 of the 21 introns in two sires (Piedmontese x Angus located at the U.S. Meat Animal Research Center in Clay Center, NE; Jersey x Limousin located at AgResearch in New Zealand) of independent resource populations previously shown to be segregating meat tenderness QTL on BTA29. The majority of the 38 SNP were found in introns or were synonymous substitutions in the coding regions, with two exceptions. Exons 14 and 9 contained SNP that were predicted to alter the protein sequence by the substitution of isoleucine for valine in Domain III of the protein, and alanine for glycine in Domain II of the protein. The resource populations were genotyped for these two SNP in addition to six intronic polymorphisms and two silent substitutions. Analysis of genotypes and shear force values in both populations revealed a difference between paternal CAPN1 alleles in which the allele encoding isoleucine at position 530 and glycine at position 316 associated with decreased meat tenderness (increased shear force values) relative to the allele encoding valine at position 530 and alanine at position 316 (P < 0.05). The association of maternal alleles with meat tenderness phenotypes is consistent with the hypothesis of CAPN1 as the gene underlying the QTL effect in two independent resource populations and presents the possibility of using these markers for selective breeding to reduce the numbers of animals with unfavorable meat tenderness traits. PMID- 12542148 TI - Estimates of genetic parameters and genetic change for reproduction, weight, and wool characteristics of Columbia sheep. AB - Genetic parameters from both single-trait and bivariate analyses for prolificacy, weight and wool traits were estimated using REML with animal models for Columbia sheep from data collected from 1950 to 1998 at the U.S. Sheep Experiment Station (USSES), Dubois, ID. Breeding values from both single-trait and seven-trait analyses calculated using the parameters estimated from the single-trait and bivariate analyses were compared with respect to genetic trends. Number of observations were 31,401 for litter size at birth and litter size at weaning, 24,741 for birth weight, 23,903 for weaning weight, 29,572 for fleece weight and fleece grade, and 2,449 for staple length. Direct heritability estimates from single-trait analyses were 0.09 for litter size at birth, 0.06 for litter size at weaning, 0.27 for birth weight, 0.16 for weaning weight, 0.53 for fleece weight, 0.41 for fleece grade, and 0.55 for staple length. Estimate of direct genetic correlation between littersize at birth and weaning was 0.84 and between birth and weaning weights was 0.56. Estimate of genetic correlation between fleece weight and staple length was positive (0.55) but negative between fleece weight and fleece grade (-0.47) and between staple length and fleece grade (-0.70). Estimates of genetic correlations were positive but small between birth weight and litter size traits and moderate and positive between weaning weight and litter size traits. Fleece weight was lowly and negatively correlated with both litter size traits. Fleece grade was lowly and positively correlated with both litter size traits, while staple length was lowly and negatively correlated with the litter size traits. Estimates of correlations between weight traits and fleece weight were positive and low to moderate. Estimates of correlations between weight traits and fleece grade were negative and small. Estimates of correlations between staple length and birth weight (0.05) and weaning weight were small (-0.04). Estimated breeding values averaged by year of birth from both the single-trait and multiple-trait analyses for the prolificacy and weight traits increased over time, but were unchanged for the wool traits. Estimated changes in breeding values over time did not differ substantially for single trait and multiple-trait analyses, except for traits highly correlated with another trait that was responding to selection. PMID- 12542149 TI - Genetic parameters for pork carcass components. AB - Data from 456 homozygous halothane normal purebred Yorkshire, Duroc, and Other breed pigs from two national progeny testing and genetic evaluation programs were utilized to estimate genetic parameters for carcass components in pigs. Carcass components were cut and weighed according to Institutional Meat Purchase Specifications. Primal cut weights evaluated included 401 Ham (HAM), 410 Loin (LOIN), 405 Picnic shoulder (PIC), 406 Boston Butt (BB), and 409 Belly (BELLY). Individual muscle weights included the inside (INS), outside (OUT), and knuckle (KNU) muscles of the ham, the longissimus dorsi (LD) and psoas major (TEND) of the loin, and the boneless components of both the Boston Butt (BBUTT) and picnic (BPIC). Muscle weights from each primal were summed to yield a boneless subprimal weight (BHAM, BLOIN, BSHLDR), and all boneless subprimals were summed to yield total primal boneless lean (LEAN). Heritability estimates for HAM, LOIN, and BELLY were 0.57, 0.51, and 0.51, respectively. Heritability estimates for BB and PIC were 0.09 and 0.21, respectively. Heritability estimates for the boneless components of each primal were higher than those for the intact primals. Genetic correlations for HAM, LOIN, and PIC with loin muscle area (LMA) were 0.53, 0.78, and 0.70, respectively, and-0.62, -0.51, and -0.60, respectively, with 10th rib off-midline backfat (BF10). Boneless subprimal components were highly correlated with LEAN. Gilts had heavier weights (P < 0.01) than barrows for all boneless subprimals, individual muscles, LEAN, and for all primal cuts except BELLY. Gilts also had less BF10 and more LMA (P < 0.01) than barrows. Duroc pigs had a heavier (P < 0.01) weight for HAM and PIC when compared to Yorkshires. Yorkshire pigs had more (P < 0.01) LOIN weight than did the Durocs. Results suggest primal, boneless subprimal, and individual muscle weights in pigs should respond favorably to selection. PMID- 12542150 TI - Estimates of parameters between direct and maternal genetic effects for weaning weight and direct genetic effects for carcass traits in crossbred cattle. AB - Estimates of heritabilities and genetic correlations were obtained for weaning weight records of 23,681 crossbred steers and heifers and carcass records from 4,094 crossbred steers using animal models. Carcass traits included hot carcass weight; retail product percentage; fat percentage; bone percentage; ribeye area; adjusted fat thickness; marbling score, Warner-Bratzler shear force and kidney, pelvic and heart fat percentage. Weaning weight was modeled with fixed effects of age of dam, sex, breed combination, and birth year, with calendar birth day as a covariate and random direct and maternal genetic and maternal permanent environmental effects. The models for carcass traits included fixed effects of age of dam, line, and birth year, with covariates for weaning and slaughter ages and random direct and maternal effects. Direct and maternal heritabilities for weaning weight were 0.4 +/- 0.02 and 0.19 +/- 0.02, respectively. The estimate of direct-maternal genetic correlation for weaning weight was negative (-0.18 +/- 0.08). Heritabilities for carcass traits of steers were moderate to high (0.34 to 0.60). Estimates of genetic correlations between direct genetic effects for weaning weight and carcass traits were small except with hot carcass weight (0.70), ribeye area (0.29), and adjusted fat thickness (0.26). The largest estimates of genetic correlations between maternal genetic effects for weaning weight and direct genetic effects for carcass traits were found for hot carcass weight (0.61), retail product percentage (-0.33), fat percentage (0.33), ribeye area (0.29), marbling score (0.28) and adjusted fat thickness (0.25), indicating that maternal effects for weaning weight may be correlated with genotype for propensity to fatten in steers. PMID- 12542151 TI - Genetic parameters for weight, weight adjusted for body condition score, height, and body condition score in beef cows. AB - Weight (CW, n = 61,798), weight adjusted for condition score (WA), hip height (CH, n = 56,494), and condition score (CS, n = 61,434) of cows (2 through 8 yr of age) produced by crosses of 22 sire breeds with Angus and Hereford dams in the first four cycles of the Germplasm Evaluation (GPE) Program at the U.S. Meat Animal Research Center were used to estimate genetic parameters with REML. The model included sire breed, dam breed, age in years, season of measurement (1 to 4) and their interactions, and year of birth and pregnancy-lactation code (PL) as fixed effects for CW and CS. The model for CH excluded PL. Random effects were additive genetic and permanent environmental effects. Univariate analyses of all data, by season and by year of age, bivariate analyses between pairs of seasons and ages (2 to 6), and between traits were done. Estimates of heritability and repeatability over all ages were 0.49, 0.54, 0.68, and 0.16, and 0.65, 0.67, 0.75, and 0.30 for CW, WA, CH, and CS, respectively. Corresponding estimates for each age and season were similar for all traits and cycles. Estimates of genetic and permanent environmental correlations were close to unity for all pairs of seasons and traits. Genetic correlations were greater than 0.92 for all pairs of ages for CW, WA, and CH, and greater than 0.67 for CS. Genetic correlations were 0.80, 0.86, 0.43, and -0.04 for CW-CH, WA-CH, CW-CS, and CH-CS, respectively. Results suggest that repeatability models can be used to model weights and heights in this population. PMID- 12542152 TI - Breed comparisons of Angus, Charolais, Hereford, Jersey, Limousin, Simmental, and South Devon for weight, weight adjusted for body condition score, height, and body condition score of cows. AB - Breed means and differences for weight (CW, n = 19,851), height (CH, n = 14,553), and condition scores (CS, n = 19,536) recorded in four seasons per year were evaluated for 881 cows ranging from 2 to 7 yr of age from Cycle I of the Germplasm Evaluation Program at the U.S. Meat Animal Research Center. Cows were straightbred Herefords and Angus and topcrosses from mating of Hereford, Angus, South Devon, Jersey, Simmental, Limousin, and Charolais sires to Hereford and Angus dams. The model included cow age, season of measurement, and their interactions, with year of birth, pregnancy-lactation (PL) code, and breed group as fixed effects for CW and CS. Analyses of weight adjusted for condition score included CS as covariate. Model for CH excluded PL. Random effects were additive genetic and permanent environmental effects. Data were analyzed by REML. Differences due to breeds of sire were significant for all traits. Differences were generally maintained across ages, with few interchanges in ranking through maturity. Rankings were in the following order: Jersey (lightest and shortest), Hereford-Angus (and reciprocal), Limousin, South Devon, Simmental, and Charolais (heaviest and tallest). The only exception was that Limousin-sired cows were heavier than South Devon-sired cows after 5 yr of age. Cows sired by breeds of British origin tended to be lighter than breeds of continental European origin. Adjustment for condition score changed estimates of breed differences. Rankings of breed groups, however, were generally the same for actual weight and weight adjusted for condition score. Results indicated that the part of the differences in weight due to differences in condition were of small magnitude. Differences tended to increase when adjusted for condition score, especially in contrasts of continental vs British breeds. Differences among breed groups for height followed differences for weight closely. PMID- 12542153 TI - Comparisons of Angus-, Braunvieh-, Chianina-, Hereford-, Gelbvieh-, Maine Anjou-, and Red Poll-sired cows for weight, weight adjusted for body condition score, height, and body condition score. AB - Data from Angus, Hereford, and top-cross cows (n = 641) from 2- to 8-yr-old daughters of seven breeds of sires included in Cycle II of the Germplasm Evaluation Program at the U.S. Meat Animal Research Center, comprising cow weight (CW, n = 15,698), height (CH, n = 15,676), and condition score (CS, n = 15,667), were used to estimate breed-group differences. Data were recorded in four seasons of each year (1975 to 1982). The mixed model included cow age, season of measurement, and their interactions, year of birth, pregnancy-lactation code (PL), and breed-group as fixed effects for CW and CS. Analyses of weight adjusted for condition score included CS as covariate. The model for CH excluded PL. Random effects were additive genetic and permanent environmental effects. Differences among breed-groups were significant for all traits at different ages and were maintained across ages, with few interchanges in ranking through maturity. Cows were ranked (by breed of sire) in the following order for weight: Red Poll (lightest), Hereford-Angus (reciprocal), Braunvieh, Gelbvieh, Maine Anjou, and Chianina (heaviest). In general, cows sired by breeds of British origin were lighter and shorter than those of continental origin. Differences in weight due to differences in condition seemed to be of small magnitude because making an adjustment for condition score did not affect rankings of breed groups across ages. Differences among breed groups for height were consistent with differences for weight. Cows from Chianina sires were taller than Hereford-Angus cows by 14 to 15 cm across ages. In this study, breed of sire effects were significantly different for the mature size of their daughters. PMID- 12542154 TI - Breed comparisons of Angus, Brahman, Hereford, Pinzgauer, Sahiwal, and Tarentaise for weight, weight adjusted for condition score, height, and body condition score. AB - Data from topcross cows (n = 468) from six breeds of sire (Angus, Brahman, Hereford, Pinzgauer, Sahiwal, Tarentaise) and two breeds of dam (Angus and Hereford) of Cycle III of the Germplasm Evaluation (GPE) program at the U.S. Meat Animal Research Center (MARC) comprising cow weight (CW, n = 9,012), height (CH, n = 9,010), and condition score (CS, n = 8,991) recorded in four seasons per year from 2 to 6 yr of age were used to estimate breed-group differences. The mixed models included cow age, season of measurement and their interactions, year of birth, pregnancy-lactation code (PL), and breed group as fixed effects for CW and CS. Analyses of weight adjusted for condition score included CS as a linear covariate. Model for CH excluded PL. Random effects were additive genetic and permanent environmental effects. Differences among breed-groups were significant for all traits for different ages and were maintained across ages, with few interchanges in ranking through maturity. Cows with Sahiwal sires were lightest (392 to 479 kg), whereas Hereford-Angus (HA) reciprocal-cross cows were shortest (119 to 123 cm) at each age. Cows with Brahman sires were heaviest and tallest among breed groups at all ages, exceeding HA cows by 19 to 24 kg and 9 to 10 cm, respectively. Cows with Pinzgauer and Tarentaise sires were intermediate for weight and height and interchanged ranking across ages. Differences in weight due to differences in condition seemed to be of small magnitude because adjustment for condition score did not affect rankings of breed groups across ages. Important changes for mature size of cows can be achieved by breed substitution with the breeds of sires used in this study. PMID- 12542155 TI - Genetic parameter estimates of yearling live animal ultrasonic measurements in Brangus cattle. AB - The objective of this study was to estimate genetic parameters for real-time ultrasound measurements of longissimus muscle area (LMA), 12th rib backfat thickness (FT), percent intramuscular fat (IMF), and yearling weight (YW) for 1,299 yearling Brangus bulls and heifers. A single ultrasound technician performed all measurements. The number of observations was 1,298, 1,298, 1,215, and 1,170 for LMA, FT, IMF, and YW, respectively. Genetic parameters were estimated for each trait using single- and multiple-trait derivative-free restricted maximal likelihood. Fixed effects were contemporary group (defined as same sex, same age within six months, and same environment), and days of age as a covariate. Correlations were estimated from two-trait models. Heritabilities for LMA, FT, IMF, and YW were 0.31, 0.26, 0.16, and 0.53, respectively. Genetic correlations between LMA and FT, LMA and IMF, LMA and YW, FT and IMF, FT and YW, and IMF and YW were 0.09, 0.25, 0.44, 0.36, 0.42, and 0.31, respectively. Yearling live animal ultrasonic measurements can be used as a selection tool in breeding cattle for the improvement of carcass traits. PMID- 12542156 TI - Influence of macronutrients and polyethylene glycol on intake of a quebracho tannin diet by sheep and goats. AB - We determined if supplemental macronutrients or polyethylene glycol (PEG) influenced intake of a tannin diet. Sheep (lambs 5 mo age, 36 kg) and goats (kids 7 mo age, 32 kg) were fed supplements high in either energy or protein or offered a choice between the two supplements before and after receiving a meal containing 15% quebracho tannin. The effect of PEG, a compound that attenuates the negative effects of tannins, was assessed by offering PEG while animals consumed the tannin diet for 4 h/d. Intake of the tannin diet was influenced by both macronutrients and PEG. Animals that chose their own supplements or that received the high-protein supplement, consumed more of the tannin diet than animals fed the high-energy supplement: 34 and 36 vs 31 g/kg(0.75) (lambs) and 41 and 39 vs 34 g/kg(0.75) (kids), respectively (P < 0.05). Animals supplemented with PEG ate much more of the tannin diet than unsupplemented animals: 70 vs 39 g/kg(0.75) (lambs) and 63 vs 34 g/kg(0.75) (kids), respectively (P < 0.001). Sheep and goats consumed more tannin food when given PEG than when supplemented with macronutrients (51 and 38 g/kg(0.75), P < 0.001). Sheep and goats offered a choice between supplements consumed more CP than animals fed the high-energy supplement and more ME than animals fed the high-protein supplement (P < 0.05). In so doing, they selected a combination of foods that yielded a more balanced intake of macronutrients, while achieving high levels of intake of the tannin food. Sheep and goats can be used as an environmentally safe and economically sound means to reduce the abundance of tannin-rich vegetation. Macronutrients and PEG enhance use of tannin-containing plants, which may increase production of alternate forages and create a more diverse mix of species in a plant community. PMID- 12542157 TI - The relationship between meal composition and long-term diet choice. AB - When animals are offered a choice of feeds that are nutritionally complementary, they are able to select a consistent combination of these feeds over long periods of time. Analysis of how such consistent diet choice is achieved, in terms of short-term feeding behavior, may further our knowledge of how animals regulate nutrient intake. Previous work, on meal pattern analysis and on nutrient synchronization, led us to hypothesize that animals may select a consistent diet within a meal. In three experiments cows were offered a choice between high- (H) and low- (L) protein feeds and short-term feeding behavior data were collected using computerized feeders. Feeding behavior was first analyzed in terms of visit characteristics. A greater average daily intake of H, relative to L, was more closely related to the ratio of H visits to L visits than to differences in the intake per visit to feeders supplying H or L. Individual meal criteria were estimated using a mixed-distribution model, and visits were clustered into meals. Cows typically had approximately six meals per day. The observed frequency distribution of meal composition, in terms of the proportion of visits to H feeders, was determined. Subsequently, the observed visits were randomly reclustered into bouts consisting of the same number of visits as were observed in meals, and the frequency distribution of random bout composition was calculated. If frequency distributions of meals and random bouts coincide, then this is evidence that cows do not regulate diet choice within a meal. Comparison of the frequency distributions of meals and random bouts provided no evidence that cows attempted to achieve their long-term average diet composition within a meal. We also investigated whether cows tried to achieve a consistent diet choice within a meal by adjusting their intake per visit, depending on the feed type visited and the proportion of visits to H feeders in a meal. There was no evidence that this occurred. In conclusion, our analyses have shown that cows did not attempt to select within a meal a consistent diet in terms of protein to energy ratio. Indeed, our data and the literature suggest that the timeframe over which the intake of energy and protein is regulated must be greater than a meal in a number of animal species. PMID- 12542158 TI - Synthetic maternal pheromone stimulates feeding behavior and weight gain in weaned pigs. AB - One hundred and forty-four pigs were used to determine the effects of a putative synthetic maternal pheromone on behavior and performance of weanling pigs. Each pen of weaned pigs contained three pigs that were given free access to water and feed. Pigs were videotaped in time lapse for 48 h after weaning and weekly body weights and feed disappearances were recorded for 4 wk. Treatments included: a) control (vehicle applied), b) 30 mL of synthetic pheromone applied to the feeder, or c) 10 mL of synthetic pheromone applied to each of three pigs' snouts. Pigs exposed to the synthetic pheromone spent more (P < 0.05) time with their heads in the feeder and less (P < 0.05) time drinking, lying down, or engaged in agonistic behaviors than control pigs. Pigs exposed to the synthetic pheromone were more (P < 0.05) active during the 48-h period of video taping than control pigs. Pigs exposed to the synthetic pheromone (either on the feeder or their snout) had increased (P < 0.01) average daily gain (ADG) and better (P < 0.01) feed:gain ratio than control pigs over the 28-d postweaning period. In conclusion, the putative synthetic pheromone, applied once at weaning, stimulated apparent feeding behaviors, and reduced fighting and apparent drinking behaviors during the first 48 h after weaning. ADG and feed:gain ratio were improved by application of the putative synthetic pheromone either directly on the feeder or when painted on the pigs' snouts. Olfactory signals can modulate adaptation to the postweaning environment in ways that may improve pig performance and welfare. PMID- 12542159 TI - Feeding intensity and dietary protein level affect adipocyte cellularity and lipogenic capacity of muscle homogenates in growing pigs, without modification of the expression of sterol regulatory element binding protein. AB - Muscle fat stores at slaughter partly determine the dietetic and sensory quality traits of pork meat. Nutritional strategies during the growing-finishing period are able to modify intramuscular fat content; however, the underlying mechanisms remain largely unknown. The objective of this study was to determine some of the cellular, biochemical, and molecular bases of muscle fat content variation in response to feeding regimen in pigs. Crossbred pigs of 30 kg BW were allocated to three feeding groups: free access ([C], n = 10) to a standard diet (3.25 kcal of DE/kg, 9.5 g of lysine/kg), standard diet at 75% of the spontaneous voluntary intake ([FR], n = 10), or both low protein and energy intakes ([PR], n = 10) in order to get the same growth rate as the FR pigs and the same body composition as the C pigs. At slaughter (110 kg BW), FR and PR pigs were 30 d older than C pigs (P < 0.001). In agreement with the protocol, carcass adiposity was similar in PR and reduced (P < 0.01) in FR pigs compared with C animals. Lipid content in longissimus lumborum muscle was reduced by 25% in FR pigs and increased by 40% in PR pigs compared to C pigs (P < 0.001). Commensurate variations in the diameter of muscle adipocytes were observed between the three feeding groups (P < 0.001). The muscle activities of malic enzyme and glucose-6-phosphate dehydrogenase, generating reduced nicotinamide adenine dinucleotide phosphate for fatty acid synthesis, were depressed (P < 0.05) in both FR and PR groups, compared to the C group. The expression level of the sterol regulatory element binding protein that was chosen as the putative candidate at the molecular level was not modified by the feeding regimen. No variations in the oxidative enzyme markers were denoted, whereas lactate dehydrogenase activity was reduced by 13% (P < 0.05) in PR group compared to other groups. In conclusion, moderate long-term feed restriction results in decreased lipogenic capacity of muscle adipocytes and intramuscular fat content. In contrast, the reduction of both protein and energy intakes more likely results in an imbalance between multiple aspects of muscle energy metabolism, in favor of intramuscular fat accretion. PMID- 12542160 TI - Effects of fasting and transportation on pork quality development and extent of postmortem metabolism. AB - One hundred seventy-seven pigs were used to determine the interaction effects of fasting and length of transport prior to harvest on pork muscle quality. The study design was a 2 x 2 x 3 factorial, which involved two genetic sources, fasting (F) or no fasting (N) of pigs 48-h prior to harvest, and three transport times (0.5, 2.5, or 8.0 h) on a semitrailer to the packing plant. Genetic source was a significant source of variation (P < 0.05) for most composition and muscle quality variables. Fasting reduced hot carcass weight 3.6% (P < 0.05), but length of transport did not affect hot carcass weight (P > 0.05). There were no differences (P > 0.05) in percent lean among fasting and transport treatments. Fasted pigs had higher longissimus dorsi (LD) ultimate pH (pHu), darker lean color, higher marbling score and lower 7-d purge loss, 24-h drip loss, and cooking loss (P < 0.05) than nonfasted pigs. Meat from pigs that were transported 8.0 h had lower glycolytic potential (GP), higher LD and semimembranosus (SM) pHu, darker lean color, and lower L*, 7-d purge loss, 24-h drip loss, cooking loss, and shear force values than meat from pigs transported 0.5 h (P < 0.05). Meat from pigs transported 2.5 h had higher LD and SM pHu and lower L*, 7-d purge loss, 24-h drip loss, and cooking loss than meat from pigs transported 0.5 h (P < 0.05). Meat from pigs transported 8.0 h had higher LD pHu and color scores and lower L* and cooking loss than meat from pigs transported 2.5 h (P < 0.05). The fasting x transport interaction was significant for SM pHu, L*, color score, and drip loss. Fasting improved SM pHu, L*, color score, and drip loss for pigs that were transported 0.5 h (P < 0.05), but when pigs were transported for 2.5 h or 8.0 h, fasting had little or no effect on these muscle quality traits. Fasting lowered GP and increased LD pHu for pigs from the genetic source with the higher initial pork quality (P < 0.05), while fasting had no effect on pork quality for pigs from the genetic source with the lower initial pork quality (P > 0.05). Longer transport times resulted in lower GP and higher LD pHu regardless of genetic source. Fasting and length of transport each had positive effects on pork quality, but length of transport effects was greater in magnitude. When pigs were transported for 0.5 h, fasting for 48 h prior to harvest improved pork quality, but when pigs were transported 2.5 or 8.0 h, fasting had little effect on pork quality. PMID- 12542161 TI - Effects of increasing dietary niacin on growth performance and meat quality in finishing pigs reared in two different environments. AB - We conducted two experiments to determine the effects of added dietary niacin on growth performance and meat quality in finishing pigs. Pigs were blocked by weight and assigned to one of six dietary treatments in both experiments. Dietary treatments consisted of a corn-soybean meal-based control diet (no added niacin) or the control diet with 13, 28, 55, 110, or 550 mg/kg of added niacin. In Exp. 1, pigs were housed at the Kansas State University research from with two pigs per pen (six pens per treatment per sex). In Exp. 2, pigs were housed with 26 pigs per pen (four pens per treatment per sex) in a commercial research barn. In Exp. 1, 144 pigs (initially 51.2 kg) were fed diets in two phases (d 0 to 25 and 25 to 62) that were formulated to 1.00 and 0.75% lysine, respectively. In Exp. 2, 1,248 pigs (initially 35.9 kg) were fed diets in four phases (d 0 to 28, 29 to 56, 57 to 84, and 85 to 117), with corresponding total lysine concentrations of 1.25, 1.10, 0.90, and 0.65% lysine, respectively. Added fat (6.0%) was included in the first three phases. In Exp. 1, average daily feed intake tended (quadratic, P < 0.07) to increase then return to values similar to control pigs as dietary niacin increased. Longissimus muscle (LM) 24-h pH (longissimus of pigs fed added niacin) tended to increase (control vs niacin, P < 0.06) for pigs fed added niacin. In the commercial facility (Exp. 2), increasing added niacin improved gain:feed (quadratic, P < 0.01) and subjective color score, and ultimate pH (linear, P < 0.01). Added niacin also decreased (linear, P < 0.04) carcass shrink, L* values, and drip loss percentage. Results from these two studies show that 13 to 55 mg/kg added dietary niacin can be fed to pigs in a commercial environment to improve gain:feed. It also appears that pork quality, as measured by drip loss, pH, and color, may be improved by higher concentrations of added dietary niacin. PMID- 12542162 TI - Effects of growth potential and growth path on tenderness of beef longissimus muscle from bulls and steers. AB - The influence of growth potential or growth path on the tenderness of the longissimus muscle was investigated using 117 Angus and Angus-cross bulls and steers raised on pasture over two successive years. Growth rate for a period of 100 d from a weight of about 200 kg was used to identify the faster-growing two thirds of cattle within the gender groups, half of which were grown fast to a slaughter weight of 530 kg at 16 to 18 mo of age (the Fast group), whereas the other half were restricted in growth (the Restricted group) so they attained a similar final weight as the slower-growing third (the Slow group) at about 26 mo of age. The Restricted group was included to determine whether the tougher meat expected from the Slow group relative to the Fast group (based on previous results) was due to the greater age of the Slow group or to their slower early growth rate. Beef from the Fast group was tenderer than that from both the Slow and Restricted groups based on sensory panels (P < 0.05) and objective measures (P < 0.05), indicating that the early growth-rate potential was less important than the differences in age or the patterns of growth for the Slow and Restricted groups. Improved tenderness for the Fast group was associated with more intramuscular fat (P < 0.05) and higher myofibrillar fragmentation indexes (P < 0.05). Patterns of tenderness differences between treatment groups were similar for bulls and steers, but beef from bulls was tougher (P < 0.001) than that from steers. The more tender beef from steers was associated with a slightly lower ultimate pH (P < 0.001), higher myofibrillar fragmentation indexes (P < 0.001), and more intramuscular fat (P < 0.001). Ultimate pH affected beef tenderness (P < 0.01), but adjustments to a constant pH did not decrease differences between treatment and gender groups. The higher growth rates (P < 0.01) and leaner carcasses (P < 0.01) of bulls compared with steers were consistent with other studies. Increases in age of 8 to 10 mo may be associated with less tender beef for cattle finished on pasture, and beef from bulls is likely to be less tender than that from steers. PMID- 12542163 TI - Additivity and associative effects of metabolizable energy and amino acid digestibility of corn, soybean meal, and wheat red dog for White Pekin ducks. AB - The additivity of true amino acid digestibility (TAAD) and true metabolizable energy (TME) values in corn, soybean meal, and wheat red dog for White Pekin ducks was investigated. Differences between observed values for the complete diets and values predicted from measurements of individual ingredients were used to test additivity. Eight ducks were each assigned to the following dietary treatments: corn, soybean meal (44% CP), wheat red dog (wheat by-product with less than 4% fiber), complete Diet 1 (corn-soybean meal), complete Diet 2 (corn red dog-soybean meal), and dextrose. Dextrose-fed ducks were used to estimate endogenous losses. The nitrogen-corrected TME (TMEn) in corn, soybean meal, wheat red dog, and two complete diets were 3.411, 2.919, 2.502, 3.148, and 3.111 kcal/g, respectively. In general, the TME and TMEn values observed in the two complete diets were not different (P > 0.05) from predicted values and indicated that the TME and TMEn in corn, soybean meal, and wheat red dog were all additive. The mean TAAD of corn, soybean meal, wheat red dog, and the two complete diets were not different, and were 87.03, 88.15, 90.58, 85.83, and 87.02%, respectively. The differences in TAAD between observed and predicted values were significant (P < 0.05) only for arginine, lysine, and aspartate in complete Diet 1, and for arginine, histidine, lysine, and aspartate in complete Diet 2. These results indicated that TME and TMEn values for corn,soybean meal, and wheat red dog were all additive in the two complete diets, but digestibilities of some amino acids were not additive and demonstrated some associative effects. PMID- 12542164 TI - Effect of vitamin E on improving fresh pork quality in Berkshire- and Hampshire sired pigs. AB - This study was designed to evaluate the effects of vitamin E supplementation on pork quality of two genotypes with distinct differences in pork quality traits. Pigs (n = 240; BW = 87 +/- 0.35 kg) were allotted by weight to one of 20 treatments (4 pens/treatment, 3 pigs/pen) in a 2 x 2 x 5 factorial randomized complete block design. Factors included 1) genotype (Berkshire or Hampshire sired), 2) sex (gilts or barrows), and 3) vitamin E level (12.1, 54.7, 98.8, 174.0, and 350.6 IU of vitamin E/kg diet). Hampshire-sired pigs had greater average daily gain (1.05 vs 0.98 kg) and gain:feed (0.30 vs 0.27) and less average daily feed intake (ADFI) (3.46 vs 3.62 kg) than Berkshire-sired pigs (P < 0.001) for the 6-wk study. Hampshire-sired barrows consumed more feed (3.54 vs 3.38 kg/d) and were less efficient (0.29 vs 0.31) than Hampshire-sired gilts (P < 0.05), but this sex difference was not observed in Berkshire-sired pigs (interaction, P < 0.05). Berkshire-sired pigs had greater backfat (34.1 vs 21.1 mm; P < 0.001), reduced longissimus muscle area (37.6 vs 46.3 cm2; P < 0.001), reduced lean percentage (53.0 vs 55.8; P < 0.001), and a greater head-on yield (79.8 vs 79.2; P < 0.05). Vitamin E increased (P < 0.05) ADFI linearly (P < 0.05), but had no effects on carcass composition. Loin chops from Hampshire-sired pigs had reduced ultimate pH (5.64 vs 5.91), greater drip loss (92.2 vs 66.3 mg), and increased Minolta L* (52.6 vs 48.6), a* (8.9 vs 7.5), and b* (6.9 vs 5.2) values compared to Berkshire-sired pigs (P < 0.001). Vitamin E had no effect on pH, temperature, drip loss, and L* or a* values, but tended (P < 0.07) to increase b* values linearly (P < 0.06). Oxidation as indicated by thiobarbituric acid reactive substances (TBARS) was greatest in Hampshire-sired gilts at the lowest level of vitamin E, and decreased linearly (P < 0.001) with additional vitamin E. However, TBARS responded in a cubic fashion (P < 0.05) to vitamin E in Hampshire-sired barrows and were not affected in Berkshire-sired gilts or barrows (three-way interaction, P < 0.02). Hampshire-sired pigs had greater TBARS than Berkshire-sired pigs (0.053 vs 0.047 mg malondialdehyde equivalents/kg). Vitamin E supplementation increased serum concentrations of vitamin E on d 21 (1.06 to 4.79 microg/mL) and d 42 (1.02 to 2.82 microg/mL) and increased tissue concentrations of vitamin E (1.99 to 4.83 microg/g) linearly (P < 0.001). Vitamin E supplementation was not effective in improving fresh meat quality in genotypes with poor or superior meat quality traits. PMID- 12542165 TI - Effects of dietary protein and oathull fiber on nitrogen excretion patterns and postprandial plasma urea profiles in grower pigs. AB - The objectives of this study were: 1) to determine if dietary protein reduction or oathull fiber inclusion would reduce urinary N excretion in grower pigs, 2) to determine if plasma urea could predict urinary N excretion among diets differing in protein and fiber content with an expected range in N excretion patterns, and 3) to determine the postprandial time point to sample blood for the best prediction. Three dietary protein concentrations (high, 19.7; medium, 16.9; low, 13.8%) and two fiber levels (high, 5.0; low, 3.6% crude fiber) were tested in a 3 x 2 factorial arrangement. Diets (wheat, barley, soybean meal; oathulls as fiber source) were formulated to 3.25 Mcal of digestible energy (DE)/kg and 2.2 g of digestible lysine/Mcal DE for low- and medium-protein diets, and 2.4 g/Mcal of DE for high-protein diets, and supplemented with lysine, methionine, tryptophan, threonine, isoleucine, or valine to meet an ideal amino acid profile. Pigs (32 +/ 3.4 kg; n = 42) were housed in metabolism crates for 19 d. On d 10 or 11, catheters were installed by cranial vena cava venipuncture. Daily feeding allowance was adjusted to 3x maintenance (3 x 110 kcal DE/kg body weight(0.75)), and was fed in two equal meals. Feces and urine were collected from d 15 to 19. Five blood samples were collected in 2-h intervals on d 16 and 19. Fecal, urinary, and total N excretion was reduced linearly with a reduction of dietary protein (P < 0.001); the reduction was greater for urinary (48%) and total N excretion (40%) than for fecal N excretion (23%). Similarly, the ratio of urinary to fecal N was reduced linearly with a reduction of dietary protein (P < 0.001). Retention of N (g/d) was reduced linearly, but N retention as a percentage of N intake was increased linearly with a reduction of dietary protein (P < 0.001). The addition of oathulls did not affect N excretion patterns and plasma urea (P > 0.10). Dietary treatments did not affect average daily gain or feed efficiency (P > 0.10). A dietary protein x time interaction affected plasma urea (P < 0.001). For medium- and high-protein diets, plasma urea increased postprandially, peaking 4 h after feeding, and then decreased toward preprandial levels (P < 0.05). Plasma urea did not alter postprandially for the low-protein diet (P > 0.10). Urinary N excretion (g/d) was predicted by 3.03 + 2.14 x plasma urea concentration (mmol/L) at 4 h after feeding (R2 = 0.66). Plasma urea concentration is indicative of daily urinary N excretion and reduction of dietary protein is effective to reduce total and urinary N excretion. PMID- 12542166 TI - Effects of dietary protein and fermentable fiber on nitrogen excretion patterns and plasma urea in grower pigs. AB - Effects of dietary protein concentration (high, 18.5; low, 15.7%) and fermentable fiber (control; soyhulls, SH; and sugar beet pulp, SBP) on N excretion patterns and plasma urea were tested in a 2 x 3 factorial arrangement. The objectives were: 1) to determine if reduced dietary protein together with fermentable fiber would reduce urinary N excretion further than a single diet manipulation, 2) to determine if effects of diet manipulations were similar between pigs with restricted and free access of feed, and 3) to further develop predictions of urinary N excretion using plasma urea. Diets were formulated to 3.30 Mcal digestible energy (DE)/kg and 2.4 g of digestible lysine per Mcal DE, and supplemented with lysine, methionine, tryptophan, threonine, isoleucine, leucine, or valine to ensure meeting an ideal AA profile. Pigs (30.5 +/- 3 kg; n = 36) were housed in metabolism crates with restricted access to feed (3 x 110 kcal DE/kg BW(0.75)) from d 1 to 18, and free access from d 19 to 26. Feces and urine were collected from d 15 to 18 and d 23 to 26, and blood was sampled on d 17 and 25. With restricted access to feed, urinary N was reduced 28% and N retention was reduced 12% for the low- compared to high-protein diet (P < 0.01; as g/d). Fecal N was increased 4% units for SH and 6.5% units for SBP (P < 0.01; as % of N intake) and urinary N was reduced 5% units for SH (P < 0.10) and 9% units for SBP (P < 0.05) compared to the control. With free access to feed, urinary N was reduced 27% (P < 0.05; as g/d) and N retention was reduced 7% (P < 0.10) for the low- compared to high-protein diet. Fecal N was increased 5% units for SH and 9% units for SBP (P < 0.001; as % of N intake), and urinary N was reduced 9% units for SH and 10% units for SBP (P < 0.01) compared to the control. For either restricted or free access to feed, fermentable fiber did not affect N retention (P > 0.10). A protein x fiber interaction was not observed for urinary N excretion (P > 0.10), indicating that reducing dietary protein and including fermentable fiber reduced urinary N excretion in an additive manner. Daily urinary N excretion was related positively and linearly with plasma urea in pigs with free access to feed (R2 = 0.71; at 0800). In summary, reduction of dietary protein reduced urine N excretion, and fermentable fiber shifted N excretion from urine to feces. Effects of dietary protein and fermentable fiber on reducing urinary N excretion are additive. PMID- 12542167 TI - Effects of feeding a blend of grains naturally contaminated with Fusarium mycotoxins on swine performance, brain regional neurochemistry, and serum chemistry and the efficacy of a polymeric glucomannan mycotoxin adsorbent. AB - The co-occurrence of Fusarium mycotoxins in contaminated swine diets has been shown to result in synergistic toxicity beyond that observed for individual toxins. An experiment was conducted, therefore, to investigate the effects of feeding a blend of grains naturally contaminated with Fusarium mycotoxins on growth, brain regional neurochemistry, serum immunoglobulin (Ig) concentrations, serum chemistry, hematology, and organ weights of starter pigs. Three levels of glucomannan polymer (GM polymer, extract of yeast cell wall, Alltech Inc.) were also tested for its efficacy to overcome Fusarium mycotoxicoses. A total of 175 starter pigs (initial weight of 10 +/- 1.1 kg) were fed five diets (seven pens of five pigs per diet) for 21 d. Diets included (1) control, (2) blend of contaminated grains, (3) contaminated grains + 0.05% GM polymer (4) contaminated grains + 0.10% GM polymer and (5) contaminated grains + 0.20% GM polymer. Diets containing contaminated grains averaged 5.5 ppm deoxynivalenol, 0.5 ppm 15 acetyldeoxynivalenol, 26.8 ppm fuuric acid, and 0.4 ppm zearalenone. Feed intake and weight gain of all pigs fed contaminated grains was significantly reduced compared to controls throughout the experiment. The weights of liver and kidney, expressed as a percentage of body weight, were lower in pigs fed the contaminated diet than in those fed the control diet. The feeding of contaminated grains significantly reduced concentrations of dopamine in the hypothalamus and pons and concentrations of dihydroxyphenylacetic acid and norepinephrine in the pons. The ratios of 5-hydroxyindoleacetic acid to serotonin, however, were elevated in the hypothalamus and pons. The feeding of contaminated grains increased serum IgM and IgA concentrations, while serum IgG concentrations were not altered. The supplementation of GM polymer prevented some of the mycotoxin-induced alterations in brain neurotransmitter and serum Ig concentrations. In summary, the feeding of grains naturally contaminated with Fusarium mycotoxins reduced growth, altered brain neurochemistry, increased serum Ig concentrations, and decreased organ weights in starter pigs. Some of the Fusarium mycotoxin-induced changes in neurochemistry and serum Ig concentrations can be prevented by the feeding of yeast cell wall polymer at appropriate concentrations, although this was not reflected in increased growth rate under these experimental conditions. PMID- 12542168 TI - Influence of exogenous gonadotropin-releasing hormone on ovarian function in beef cows after short- and long-term nutritionally induced anovulation. AB - The effect of pulsatile infusion of gonadotropin-releasing hormone (GnRH) on follicular function was evaluated in nutritionally induced anovulatory beef cows. After 4 (short; n = 12) or 18 wk (long; n = 12) of anovulation, cows were randomly assigned within anovulatory group to either 2 microg of GnRH treatment or saline (control; i.v.) every hour for 5 d. Ovarian structures were monitored by daily ultrasonography. Growth rate of the largest follicle (P < 0.01) and maximal size of the largest follicle during treatment were greater (P < 0.01) for GnRH vs control cows. At exsanguination after 5 d of GnRH treatment, the size of the second-largest follicle was greater (P < 0.05) in short (i.e., 4 wk) anovulatory cows than in long (i.e., 18 wk) anovulatory cows and the largest follicle tended (P < 0.10) to be larger in long vs short anovulatory cows. Short anovulatory GnRH-treated cows had more small follicles than short anovulatory control cows or long anovulatory GnRH-treated or control cows (anovulation x GnRH; P < 0.10). Follicular fluid (FFL) concentrations of estradiol (P < 0.01) and androstenedione (P < 0.05) were greater in GnRH vs control cows. Concentrations of insulin-like growth factor-I were greater (P < 0.10) in large vs small follicles in cows that were anovulatory for 4 wk, but not in cows that were anovulatory for 18 wk. The amount of insulin-like growth factor-binding protein (IGFBP)-3 in FFL was greater (P < 0.05) in 4- vs 18-wk anovulatory cows. Amounts of IGFBP-2, -4, and -5 were greater (P < 0.001) in FFL of small (< 5 mm) vs large (> or = 5 mm) follicles regardless of treatment. We conclude that pulsatile treatment with GnRH for 5 d stimulates similar growth of the largest follicles in short- and long-term anovulatory beef cows, and that the duration of anovulation is not a major factor that limits follicular growth w hen anovulatory cowsare treated with GnRH. The primary intrafollicular factors associated with increased follicular size were increased concentrations of estradiol, progesterone, and insulin-like growth factor-I,and decreased concentrations of IGFBP-2, -4, and -5. Increased duration of anovulation was associated with decreased concentrations of IGF-I and IGFBP-3 in FFL. PMID- 12542169 TI - High versus low body condition in mares: interactions with responses to somatotropin, GnRH analog, and dexamethasone. AB - Mares that had previously been fed to attain body condition scores (BCS) of 7.5 to 8.5 (high) or 3.0 to 3.5 (low) were used to determine the interaction of BCS with the responses to 1) administration of equine somatotropin (eST) daily for 14 d beginning January 20 followed by administration of GnRH analog (GnRHa) daily for 21 d and 2) 4-d treatment with dexamethasone later in the spring when mares in low BCS had begun to ovulate. The majority of mares with high BCS continued to cycle throughout the winter, as evidenced by larger ovaries (P < 0.002), more corpora lutea (P < 0.05), greater progesterone concentrations during eST treatment (P < 0.04), and more (P < 0.05) large- and medium-sized follicles. Treatment with eST alone or in combination with GnRHa had no effect (P > 0.05) on ovarian activity or ovulation. Plasma leptin concentrations were greater (P < 0.002) in mares with high BCS; however, there was no effect (P > 0.10) of eST treatment. Plasma IGF-I concentrations were greater (P < 0.0001) in mares treated with eST compared with mares given vehicle, and mares with high BCS had greater IGF-I (P < 0.02) and LH concentrations (P < 0.02) than mares with low BCS. Plasma leptin concentrations in mares with high BCS were increased (P < 0.001) within 12 h of dexamethasone treatment; the leptin response (P < 0.001) in mares with low BCS was greatly reduced (P < 0.001) and transient. Glucose and insulin concentrations also increased (P < 0.0001) after dexamethasone treatment in both groups, and the magnitude of the response was greater (P < 0.0001) in mares with high BCS than in mares with low BCS. In summary,low BCS in mares was associated with a consistent seasonal anovulatory state that was affected little by eST and GnRHa administration. In contrast, all but one mare with high BCS continued to experience estrous cycles and(or) have abundant follicular activity on their ovaries. The IGF-I response to eST treatment was also reduced in mares with low BCS, as was the basal leptin concentration and leptin response to dexamethasone. Although low BCS and leptin concentrations were associated with inactive ovaries during winter and early spring, mares with low BCS eventually ovulated in April and May while leptin concentrations remained low. PMID- 12542170 TI - Protein (lysine) restriction in primiparous lactating sows: effects on metabolic state, somatotropic axis, and reproductive performance after weaning. AB - Low protein intake during lactation has been demonstrated to increase the loss of body protein and to reduce the reproductive performance of female pigs. The objectives of the current experiment were 1) to determine whether protein (lysine) restriction alters levels of somatotropic hormones, insulin, follicle stimulating hormone, and leptin around weaning, and 2) to evaluate the relationships between these eventual alterations and postweaning reproductive performance. One day after farrowing, crossbred primiparous sows were randomly allocated to one of two diets containing 20% crude protein and 1.08% lysine (C, n = 12) or 10% crude protein and 0.50% lysine (L, n = 14) during a 28-d lactation. Diets provided similar amounts of metabolizable energy (3.1 Mcal/kg). Feed allowance was restricted to 4.2 kg/d throughout lactation, and litter size was standardized to 10 per sow within 5 d after farrowing. Catheters were fitted in the jugular vein of 21 sows around d 22 of lactation. Serial blood samples were collected 1 d before (day W - 1) and 1 d after (day W + 1) weaning, and single blood samples were collected daily from weaning until d 6 postweaning (day W + 6). Sows were monitored for estrus and inseminated. They were slaughtered at d 30 of gestation. During lactation, litter weight gain was similar among treatment groups. Reduced protein intake increased (P < 0.001) sow weight loss (-30 vs -19 kg) and estimated protein mobilization throughout lactation (-4.1 vs -2.0 kg). On day W - 1, L sows had higher (P < 0.02) plasma glutamine and alanine concentrations, but lower (P < 0.05) plasma tryptophan and urea than C sows. Mean and basal plasma GH were higher (P < 0.001), whereas plasma IGF-I and mean insulin were lower in L than in C sows on day W - 1. Preprandial leptin did not differ between treatments on day W - 1, but was higher (P < 0.01) in L sows than in C sows on day W + 1. Mean FSH concentrations were similar in both treatments on day W - 1 (1.3 ng/mL), but L sows had greater (P < 0.001) mean FSH on day W + 1 than C sows (1.6 vs 1.2 ng/mL). The weaning-to-estrus interval (5 +/- 1 d) was similar in both groups. Ovulation rate was lower in L than in C sows (20.0 +/- 1 vs 23.4 +/- 1, P < 0.05). No obvious relationships between reproductive traits and metabolic hormone data were observed. In conclusion, these results provide evidence that protein (lysine) restriction throughout lactation alters circulating concentrations of somatotropic hormones and insulin at the end of lactation and has a negative impact on postweaning ovulation rate. PMID- 12542171 TI - Effects of four mono- and sesquiterpenes on the consumption of alfalfa pellets by sheep. AB - Effects of individual terpenes on alfalfa pellet intake by lambs were examined in four experiments. Forty-five lambs (nine lambs/treatment) were individually fed alfalfa pellets sprayed with either camphene, myrcene, caryophyllene oxide, or beta-pinene at one of five concentrations (one terpene per experiment). Treatments (0, 0.5, 1, 2, and 10x) were multiples of the concentration (x) of a specific terpene in tarbush. Terpenes were applied to alfalfa pellets (0.64 kg x lamb(-1) x d(-1), DM basis), and consumption was measured during a 20-min interval for 5 d. Lambs were maintained and fed alfalfa pellets in one group (except during 20-min tests) at a mean total daily intake of 4.7% of BW (DM basis). Camphene and caryophyllene oxide tended to decrease intake (linear contrasts were P = .0651 and P = .0504, respectively), whereas myrcene and beta pinene exerted no effect on the consumption of alfalfa pellets by lambs. Camphene and caryophyllene oxide may be involved in the differential herbivory of individual tarbush plants by livestock. PMID- 12542172 TI - Binding affinity and capacities for ytterbium(3+) and hafinum(4+) by chemical entities of plant tissue fragments. AB - The binding affinity of ytterbium (Yb3+) and hafinum (Hf4+) to ligands of chemical entities of fragments of bermudagrass tissues and their resistance to exchanging Yb with other ligands and to displacement by protons were investigated. Chemical entities of acid resistant NDF (ARNDF), 0.1 N acid detergent fiber (0.1 N ADF), and permanganate cellulose (CELL) were prepared from fragments of bermudagrass hay (Cynodon dactylon [L.] Pers.) obtained by grinding to pass a 2-mm sieve. 175Ytterbium and Yb, as YbCl3, were initially bound to each preparation by soaking for 12 h in pH 5.5 borate buffer to obtain Yb bound onto ligands having affinity constants for Yb equal to or greater than that for the weakly stable borate ligand, Yb > or = borate. The fraction of Yb > or = borate was measured and fragments then sequentially exposed to acetate, citrate, nitrotriacetate (NTA), and EDTA ions to allow exchange of Yb from Yb > or = borate with ligands having affinity constants for Yb equal to or greater than acetate (Yb > or = acetate), citrate (Yb > or = citrate), NTA (Yb > or = NTA), and EDTA (Yb > or = EDTA) ions. Binding of Yb > or = borate indicated the existence of two species of ligands: strong ligands binding essentially 100% of added Yb at levels of 1 to 1,300 ppm (0.1 N ADF) and at 1 to 7,000 ppm (ARNDF); and weaker ligands binding 4 and 8% of the Yb, respectively, at levels of added Yb greater than 1,300 ppm and 7,000 ppm. Ytterbium > or = acetate of ARNDF, but not 0.1 N ADF, was as resistant to exchange as Yb > or = citrate. Ytterbium > or = borate was exchanged extensively (85% or greater) with soluble ligands having affinity constants > or = NTA. Ytterbium resistance to proton displacement at pH of 1.5 increased with Yb > or = EDTA > Yb > or = NTA > Yb > or = citrate > Yb > or = acetate. Very efficient binding of Yb to CELL suggested that such chemical preparations are not representative of native cellulose. Hafnium (4+) was strongly bound to plant tissues rendering both Hf and Hf-bound DM insoluble at a pH of 1.5 and insoluble in a modified NDF solvent without EDTA. It is concluded that Yb specifically applied as Yb > or = acetate and Hf4+ are indelible markers for estimating sojourn time of undigested plant tissues at the normal pH of the rumen. Because of its resistance to proton displacement, Hf4+ would be an indelible marker for estimating sojourn time in more acidic postgastric segments of the gastrointestinal tract. PMID- 12542173 TI - The efficacy of three objective systems for identifying beef cuts that can be guaranteed tender. AB - The objective of this study was to determine the accuracy of three objective systems (prototype BeefCam, colorimeter, and slice shear force) for identifying guaranteed tender beef. In Phase I, 308 carcasses (105 Top Choice, 101 Low Choice, and 102 Select) from two commercial plants were tested. In Phase II, 400 carcasses (200 rolled USDA Select and 200 rolled USDA Choice) from one commercial plant were tested. The three systems were evaluated based on progressive certification of the longissimus as "tender" in 10% increments (the best 10, 20, 30%, etc., certified as "tender" by each technology; 100% certification would mean no sorting for tenderness). In Phase I, the error (percentage of carcasses certified as tender that had Warner-Bratzler shear force of > or = 5 kg at 14 d postmortem) for 100% certification using all carcasses was 14.1%. All certification levels up to 80% (slice shear force) and up to 70% (colorimeter) had less error (P < 0.05) than 100% certification. Errors in all levels of certification by prototype BeefCam (13.8 to 9.7%) were not different (P > 0.05) from 100% certification. In Phase I, the error for 100% certification for USDA Select carcasses was 30.7%. For Select carcasses, all slice shear force certification levels up to 60% (0 to 14.8%) had less error (P < 0.05) than 100% certification. For Select carcasses, errors in all levels of certification by colorimeter (20.0 to 29.6%) and by BeefCam (27.5 to 31.4%) were not different (P > 0.05) from 100% certification. In Phase II, the error for 100% certification for all carcasses was 9.3%. For all levels of slice shear force certification less than 90% (for all carcasses) or less than 80% (Select carcasses), errors in tenderness certification were less than (P < 0.05) for 100% certification. In Phase II, for all carcasses or Select carcasses, colorimeter and prototype BeefCam certifications did not significantly reduce errors (P > 0.05) compared to 100% certification. Thus, the direct measure of tenderness provided by slice shear force results in more accurate identification of "tender" beef carcasses than either of the indirect technologies, prototype BeefCam, or colorimeter, particularly for USDA Select carcasses. As tested in this study, slice shear force, but not the prototype BeefCam or colorimeter systems, accurately identified "tender" beef. PMID- 12542174 TI - Evaluation of finishing performance, carcass characteristics, acid-resistant E. coli and total coliforms from steers fed combinations of wet corn gluten feed and steam-flaked corn. AB - Crossbred beef steers (n = 615) were used in a 152-d experiment to compare steam flaked corn (SFC) diets containing 0, 30, or 60% wet corn gluten feed (WCGF). On d 114 to 118, ruminal and fecal samples were collected from 180 steers and analyzed for pH, VFA, and total and acid-resistant Escherichia coli and coliforms. Acid resistance of E. coli and coliform populations was determined by exposure of the samples for 1 h in pH 2, 4, and 7 citric acid/sodium phosphate buffers. Increasing levels of WCGF linearly decreased total ruminal VFA (P = 0.01) and total fecal VFA (P = 0.06), but linearly increased ruminal and fecal acetate:propionate (P < 0.01) ratio and ruminal and fecal pH (P < 0.05). Feeding increasing WCGF levels resulted in a quadratic response (P < 0.05) with respect to numbers of ruminal E. coli and total coliform populations resistant to pH 4 exposure. Steers fed 30% WCGF had higher (0.7 log units) ruminal E. coli and total coliforms after exposure at pH 4 compared to steers fed 0 or 60% WCGF. Populations of E. coli and total coliforms at pH 2 and 7 were similar for all dietary treatments. Dietary WCGF linearly increased DMI (P = 0.07) and liver abscesses (P = 0.03) and linearly decreased dietary NEg (P = 0.02). Average daily gain and feed efficiencies were greatest when steers were offered 30% WCGF (quadratic, P < 0.05). Dietary manipulations that reduce acid concentrations may not correspond to changes in acid resistance of E. coli and total coliform populations detected in the gastrointestinal tracts of cattle. Moderate levels of WCGF complement SFC finishing diets. PMID- 12542175 TI - Effects of oscillating dietary protein on ruminal fermentation and site and extent of nutrient digestion in sheep. AB - Eight cannulated wethers (BW = 52.5 +/- 5.7 kg) were used in a replicated 4 x 4 Latin square designed experiment to evaluate the effects of oscillating dietary protein concentrations on ruminal fermentation, site and extent of digestion, and serum metabolite concentrations. Four treatments consisted of a 13, 15, or 17% CP diet fed daily or a regimen in which dietary CP was oscillated between 13 and 17% on a 48-h basis (ACP). All diets consisted of 65% bromegrass hay (10.5% CP, 61.9% NDF, 37.2% ADF) plus 35% corn-based supplement and were formulated to contain the same amount of degradable intake protein (9.6% of DM) plus additional undegradable intake protein (SoyPLUS, West Central Cooperative, Ralston, IA) to accomplish CP levels above 13%. Each of four experimental periods were 16 d in duration with 12 d for diet adaptation followed by 4 d for sample collection. All wethers were fed at 3.0% of initial BW (DM basis) throughout the experiment, resulting in an average organic matter intake of 1.39 kg/d across treatments. When compared to the 15% CP daily treatment, feeding ACP had no effect (P > or = 0.10) on ruminal or lower tract N, NDF, ADF, or OM digestion. True ruminal OM digestion responded quadratically (P = 0.07) to increasing dietary CP, reaching a maximum of 52.0% of OM intake with the 15% CP treatment. Sheep fed ACP tended to have lower (P = 0.08) ruminal NH3 N concentrations and an overall higher (P = 0.0001) molar proportion of acetate compared to those fed 15% CP daily. Total VFA concentrations were not affected (P > or = 0.45) by increasing dietary CP. Microbial efficiency did not differ (P > or = 0.55); thus, bacterial N flow at the duodenum responded quadratically (P = 0.04) to increasing dietary CP. Nonbacterial N (P = 0.001) and total N (P = 0.01) flows at the duodenum and total tract N digestibility (P < or = 0.04) increased linearly as dietary CP increased. Wethers fed ACP maintained a lower (P = 0.002) serum glucose and lower (P = 0.0006) serum urea N compared to those fed 15% CP daily. Because the CP content of the diet was increased at the expense of corn, the response to increased CP observed in this experiment is most likely due to negative associative effects of supplemental starch on ruminal fermentation and microbial growth. Oscillating the CP content of the diet on a 48-h basis has little effect on digestion or N utilization in sheep compared with feeding the same quantity of protein on a daily basis. PMID- 12542176 TI - Factors affecting conjugated linoleic acid and trans-C18:1 fatty acid production by mixed ruminal bacteria. AB - The objective of this study was to identify environmental factors that influence conjugated linoleic acid (CLA) and trans-C18:1 fatty acid production by mixed ruminal bacteria. Ruminal contents were collected from a 600-kg ruminally fistulated Hereford steer maintained on pasture. Mixed ruminal bacteria were obtained by differential centrifugation under anaerobic conditions and added to a basal medium that contained a commercial emulsified preparation of soybean oil and a mixture of soluble carbohydrates (cellobiose, glucose, maltose, and xylose). Culture samples were collected from batch culture incubations at 0, 2, 4, 6, 8, 12, 24, 26, 28, 30, 32, and 48 h. Continuous culture incubations were conducted at dilution rates of 0.05 and 0.10 h(-1) with extracellular pH values of 5.5 and 6.5, and 0.5 and 1.0 g/L of mixed soluble carbohydrates. Culture samples were obtained from the culture vessel once steady-state conditions had been achieved. In batch culture, trans-C18:1 concentrations increased over time and reached a maximum at 48 h. Little CLA was produced during the first 8 h, but cis-9, trans-11 CLA concentrations remained high between 24 and 30 h. When mixed ruminal bacteria were maintained in continuous culture on 0.5 g/L of mixed soluble carbohydrates, concentrations of trans-C18:1 and cis-9, trans-11 CLA were reduced (P < 0.05) at a dilution rate of 0.05 h(-1) and an extracellular pH of 5.5. Similar effects were also observed when 1.0 g/L of mixed soluble carbohydrates was used. When extracellular pH was lowered to 5.0, neither trans C18:1 or CLA isomers were detected. In conclusion, our results suggest that culture pH appears to have the most influence on the production of trans-C18:1 and CLA isomers by mixed ruminal bacteria. PMID- 12542177 TI - Effect of high-oil corn or added corn oil on ruminal biohydrogenation of fatty acids and conjugated linoleic acid formation in beef steers fed finishing diets. AB - Three Angus steers (410 kg) cannulated in the proximal duodenum were used in a replicated 3 x 3 Latin square to evaluate the effects of dietary lipid level and oil source on ruminal biohydrogenation and conjugated linoleic acid (CLA) outflow. Dietary treatments included: 1) typical corn (TC; 79.2% typical corn), 2) high-oil corn (HOC; 79.2% high-oil corn), and 3) the TC diet with corn oil added to supply an amount of lipid equal to the HOC diet (OIL; 76.9% TC + 2.4% corn oil). Duodenal samples were collected for 4 d following 10-d diet adaptation periods. Data were analyzed with animal, square, period, and treatment in the model and planned, nonorthogonal contrasts were used to test the effects of dietary lipid content (TC vs HOC and OIL) and oil source (HOC vs OIL) on ruminal biohydrogenation. Intake and duodenal flow of total long-chain fatty acids were increased (P < 0.05) by over 63% for diets containing more lipid regardless of oil source. Apparent ruminal dry matter and long chain fatty acid digestibilities were not altered (P > 0.05) by dietary lipid level or oil source. Ruminal biohydrogenation of total and individual 18-carbon unsaturated fatty acids was greater (P < 0.05) for diets with higher lipid content. Biohydrogenation of oleic acid was greater (P < 0.05) for HOC than OIL, but biohydrogenation of linoleic acid was lower (P < 0.05) for HOC than OIL. Duodenal flows of palmitic, stearic, oleic, linoleic, and arachidic acids were more than 30% greater (P < 0.05) for diets containing more lipid. Flow of all trans-octadecenoic acids was greater (P < 0.05) for diets containing more lipid. Corn oil addition increased (P < 0.05) the flow of trans-10 octadecenoic acid and the trans-10, cis-12 isomer of CLA by threefold compared to feeding high-oil corn. Feeding high-oil corn or adding corn oil to typical corn rations increased intake, biohydrogenation, and duodenal flow of unsaturated long-chain fatty acids. Compared with high-oil corn diets, addition of corn oil increased duodenal flow of trans-10, trans-12 and cis-12 isomers of octadecenoic acid and the trans-10, cis-12 isomer of CLA. The amount of cis-9, trans-11 isomer of conjugated linoleic acid flowing to the duodenum was less than 260 mg/d, a value over 20 times lower than flow of trans-11 vaccenic acid indicating the importance of tissue desaturation for enhanced conjugated linoleic acid content of beef. PMID- 12542178 TI - Intestinal starch disappearance increased in steers abomasally infused with starch and protein. AB - Steers (379 +/- 10 kg) with ruminal, duodenal, and ileal cannulas were used in a 5 x 5 Latin square digestion trial to quantify and evaluate the relationship between intestinal protein supply and intestinal starch disappearance. Treatments were infusions of 0, 50, 100, 150, or 200 g/d of casein along with 1,042 g/d of raw cornstarch. Abomasal infusions were accomplished by passing tubing and a pliable retaining washer through the reticular-omasal orifice into the abomasum. Steers were fed a 93% corn silage, 7% supplement diet that contained 12% crude protein at 1.65% body weight in 12 equal portions/d. Periods lasted 17 d (12 d for adaptation, 2 d of collections, and 3 d of rest). The quantity and percentage of organic matter and protein disappearance from the small intestine increased linearly (P < 0.03) with infused casein. Greater quantities of starch disappeared with increased casein infusion (P < 0.01). The infusion of 200 g/d of casein increased small intestinal starch disappearance by 226 g/d over the control. Casein infusion did not affect the quantity or percent of organic matter, starch, or protein disappearance in the large intestine. Treatments did not change ruminal ammonia N, ruminal pH, or plasma glucose concentrations. Starch disappearance from the small intestine was increased with greater protein flow to the duodenum of steers. PMID- 12542180 TI - Revision of Darsie and Ward (1981) to include Ochlerotatus japonicus Theobald and a checklist of species referred to the genus Ochlerotatus in the Nearctic Region. AB - Breeding populations of Ochlerotatus japonicus japonicus, 1st detected in 1998, are now found in several states in the United States. To be able to identify this mosquito with the keys to the mosquitoes of North America north of Mexico, changes are proposed, especially dealing with the new genus Ochlerotatus. Also, a checklist of the genera Ochlerotatus and Aedes in the Nearctic Region is given. PMID- 12542179 TI - Critique of a dynamic model of N metabolism in the lactating dairy cow. PMID- 12542181 TI - Mosquito studies in Belize, Central America: records, taxonomic notes, and a checklist of species. AB - Data from mosquito collections made in Belize, Central America, between September 1990 and April 1993 are presented. A total of 537 collections yielding 15,139 specimens are summarized. One genus, 4 subgenera, and 31 species are recorded from Belize for the 1st time. A checklist of the 111 mosquito species now known to occur in Belize is presented. PMID- 12542182 TI - Redescription of the pupa of Culex (Melanoconion) iolambdis Dyar. AB - The pupa of Culex iolambdis is redescribed with the Belkin system of nomenclature, a table of setal branching, and an illustration. PMID- 12542183 TI - The structure and function of the larval siphon and spiracular apparatus of Coquillettidia perturbans. AB - The structure of the larval siphon and spiracular apparatus of Coquillettidia perturbans and the mechanism of attachment to roots of emergent aquatic macrophytes were examined by utilizing dissection and scanning electron microscopy. The roots of these plants contain large air-filled aerenchyma channels that larvae of Cq. perturbans pierce with their specialized siphon and spiracular apparatus to breathe. The siphon contains the spiracular apparatus, comprising the saw, postabdominal spiracles, inner spiracular teeth, and the spiracular apodeme. These are the primary structures that are utilized by larvae to pierce root tissue. Once entry is made into a root, the outer spiracular teeth open fully, anchoring the larva in place. PMID- 12542184 TI - Vector competence of three North American strains of Aedes albopictus for West Nile virus. AB - To evaluate the potential for North American (NA) Aedes albopictus to transmit West Nile virus (WN), mosquito strains derived from 3 NA sources (Frederick County, Maryland, FRED strain; Cheverly, MD, CHEV strain; Chambers and Liberty counties, Texas, TAMU strain) were tested. These strains were tested along with a previously tested strain from a Hawaiian source (OAHU strain). Mosquitoes were fed on 2- to 3-day-old chickens previously inoculated with a New York strain (Crow 397-99) of WN. All of the NA strains were competent laboratory vectors of WN, with transmission rates of 36, 50, 83, and 92% for the FRED, CHEV, OAHU, and TAMU strains, respectively. The extrinsic incubation period for WN in Ae. albopictus held at 26 degrees C was estimated to be 10 days. Based on efficiency of viral transmission, evidence of natural infection, bionomics, and distribution, Ae. albopictus could be an important bridge vector of WN in the southeastern USA. PMID- 12542185 TI - Seasonal prevalence of Culex vishnui subgroup, the major vectors of Japanese encephalitis virus in an endemic district of Andhra Pradesh, India. AB - Seasonal prevalence of members of the Culex vishnui subgroup, the major vectors of Japanese encephalitis virus (JE) in an endemic district of Andhra Pradesh, was investigated. Approximately 15,500 mosquitoes belonging to 2 genera and 5 species were collected by indoor-resting collections. The predominant species were found to be from the Cx. vishnui subgroup, which comprised 42.6% of the total collection, followed by Anopheles subpictus (40.4%), An. hyrcanus (12.8%), Cx. gelidus (3.5%), and An. barbirostris (0.8%). Mosquitoes of the Cx. vishnui subgroup were collected throughout the year, and densities of females ranged from 2.3/man-hour in January 1999 to 26.0/man-hour in December 1999. Ninety-three cases of Japanese encephalitis (27 confirmed serologically) were reported, mainly during the monsoon months of November and December. A significant positive correlation between densities of mosquitoes of the Cx. vishnui subgroup and occurrence of Japanese encephalitis cases (r = 0.765, P < 0.01) was observed. The factors responsible for transmission of JE in the endemic district are discussed. PMID- 12542186 TI - Comparison of vero cell plaque assay, TaqMan reverse transcriptase polymerase chain reaction RNA assay, and VecTest antigen assay for detection of West Nile virus in field-collected mosquitoes. AB - Mosquitoes collected during the epidemic of West Nile virus (WN) in Staten Island, NY, during 2000 were identified to species, grouped into pools of up to 50 individuals, and tested for the presence of WN by using TaqMan reverse transcriptase polymerase chain reaction (RT-PCR) to detect West Nile viral RNA, Vero cell plaque assay to detect infectious virus, and VecTest WNV/SLE Antigen Panel Assay. A total of 10,866 specimens was tested in 801 pools. Analysis of results indicated that TaqMan RT-PCR detected 34 WN-positive pools, more than either of the other techniques. The plaque assay detected 74% of the pools positive by TaqMan, and VecTest detected 60% of the pools positive by TaqMan. The VecTest assay detected evidence of West Nile viral antigen in 67% of the pools that contained live virus detected by plaque assay. A WN enzyme immunoassay performed similarly to the VecTest WN assay. Differences in performance were related to relative sensitivity of the tests. Infection rates of WN in Culex pipiens and Cx. salinarius calculated by the 3 techniques varied, but each estimate indicated a high infection rate in the population. Positive and negative attributes of each procedure, which may influence how and where they are used in surveillance programs, are discussed. PMID- 12542187 TI - Molecular divergence of the mitochondrial cytochrome oxidase II gene in three mosquitoes. AB - The cytochrome oxidase II (COII) genes in the mitochondrial DNA of 3 mosquito species (Anopheles sinensis, Aedes albopictus, and Culex quinquefasciatus) were amplified and sequenced. Both the gene order and direction of transcription were identical to those of other species of Anopheles, Aedes, and Culex. The polymerase chain reaction-amplified fragments in these mosquitoes were approximately 700 base pairs and the nucleotide sequences exhibited more than 82% similarity, whereas amino acids were more than 85% similar. The frequency of transitions was less than that of transversions. Four highly conserved segments of COII proteins are similar to those in other insects. These segments contain the major amino acid residues of cytochrome c oxidase involved in electron transport and ligand binding. PMID- 12542188 TI - Host feeding preferences of Anopheles species collected by manual aspiration, mechanical aspiration, and from a vehicle-mounted trap in the Toledo District, Belize, Central America. AB - The host-feeding patterns of Anopheles albimanus, Anopheles vestitipennis, and Anopheles punctimacula from the Toledo District in southern Belize were studied with blood-fed females that were collected by manual aspiration, a backpack aspirator, and a vehicle-mounted trap for sampling in-flight mosquito populations. Female An. vestitipennis collected from both inside and outside house walls by manual aspiration tested positive for human blood meals (88 and 67%, respectively). At increasing distances from the houses, specimens of An. vestitipennis collected from vegetation with the backpack aspirator were equally positive for human and cattle blood (44 and 43%, respectively). In contrast, 68% of the An. albimanus specimens (148) collected by backpack aspiration tested positive for cattle blood. Engorged An. vestitipennis from vehicle-mounted trap collections tested positive for cattle (108) and human (52) blood. Almost all specimens of An. albimanus from these collections were positive for cow (95%). After analyzing the data from the An. vestitipennis samples using the feeding index, the ratio of human blood to all other bloodmeal sources showed indices greater than 1. Both An. albimanus and An. punctimacula fed mostly on cattle and rarely fed on humans. Foraging ratios for the 3 Anopheles species were very similar to the feeding indexes. Ratios based on data from all collection methods showed that An. vestitipennis feeds predominately on humans. The foraging ratios for An. albimanus demonstrated consistent preferences for nonhuman hosts. As with previous studies. An. albimanus seemed to prefer cattle and pigs to almost all other host species. PMID- 12542189 TI - Heritability of wing-beat frequency in Anopheles quadrimaculatus. AB - The repeatability of male wing-beat frequency measurements of Anopheles quadrimaculatus was determined by using mosquitoes allowed free flight in a confined space. Heritability of the wing-beat frequency trait was estimated for a laboratory and a wild-strain population of An. quadrimaculatus by using free flight measurement with a parent-offspring regression of offspring on dams. Repeatability was 0.75 for free flight. Wing-beat frequency rose for the 1st day after adult emergence and then became steady. Female heritability of wing-beat frequency was 21.6% for colony and 24.0% for wild-strain mosquitoes. Male heritability was 57.2% for colony and 53.7% for wild-strain mosquitoes. Male heritability was significantly different from 0 when probabilities were combined across both populations. PMID- 12542190 TI - Larvicidal effect of the juvenile hormone mimic pyriproxyfen on Culex pipiens. AB - The larvicidal activity of the juvenile hormone mimic pyriproxyfen was evaluated against the 4th larval instar of Culex pipiens under 5 constant temperatures in the laboratory. Toxicity of this insect compound increased with temperature. The 50% lethal concentrations ranged between 0.00111 ppm at 20 degrees C and 0.00013 ppm at 32 degrees C. A similar trend was observed for the 90% lethal concentrations, which varied from 0.00379 ppm to 0.00024 ppm at the 2 temperatures, respectively. Some effects of the compound were observed on the ultrastructure of the 4th-stage larval integument, where electron micrographs revealed the destruction of procuticle lamellae, formation of cuticular vacuoles, deformed mitochondria, and destruction of nuclear envelopes and the epidermal layer, in addition to an increase in electron-dense lysosomelike bodies. PMID- 12542191 TI - The potential of intermittent irrigation for increasing rice yields, lowering water consumption, reducing methane emissions, and controlling malaria in African rice fields. AB - Rice production in sub-Saharan Africa has more than doubled in the last 3 decades and the potential to further develop rice-harvested areas is considerable. Several studies have demonstrated that the transformation of arable land into rice irrigation might create suitable habitats for large populations of disease vectors. Prominent among those are anopheline mosquitoes responsible for transmission of malaria. The method of irrigation on an intermittent basis during the rice-cropping calendar has gained renewed interest as a potentially effective malaria control strategy since the early 1980s. We review the experiences of the past 80 years with intermittent irrigation in the cultivation of rice. This method has been shown to reduce significantly the density of malaria vectors by curtailing their larval development. Furthermore, reduced methane emissions and water savings with at least equal yields were achieved in intermittently irrigated rice fields. We explore and discuss under what conditions intermittent irrigation might be beneficial in new rice-growing areas and identify steps that have to be taken to expand such programs in the future. PMID- 12542192 TI - Operational studies with Valent VectoLex WDG, Bacillus sphaericus, in three Florida Mosquito Control Districts. AB - VectoLex WDG, Bacillus sphaericus (Valent BioSciences Corporation), was evaluated in 3 Florida Mosquito Control Districts under operational conditions. Application rates were 0.5 and 1.0 lb/acre and an untreated control in each district. Study sites included woodland ponds, residential areas such as catch basins, and commercial business retention ponds. Pretreatment and posttreatment assessments were made with a standard dipper at designated dipping stations. The most prevalent mosquitoes included Culex quinquefasciatus, Culex nigripalpus, Culex erraticus, Ochlerotatus infirmatus, Anopheles crucians, Anopheles quadrimaculatus, and Psorophora spp. VectoLex WDG was effective for 24-35 days posttreatment with one application. PMID- 12542193 TI - Repellent activities of essential oils and monoterpenes against Culex pipiens pallens. AB - Essential oils of Eulcalyptus globulus, Lavender officinalis, Rosemarinus officinalis, and Thymus vulgaris were examined for their repellent activities against Culex pipiens pallens. All 4 essential oils effectively repelled adult mosquitoes on hairless mice. Essential oil of T. vulgaris (thyme) had potent repellent activity within the tested materials, with a protection rate of 91% at a concentration of 0.05% topical treatment. Thyme essential oil significantly extended the duration of protection until 3 bites by mosquitoes. With gas chromatography-mass spectrometry analysis, thyme essential oil was a rich source of 5 monoterpenes, including in descending order thymol, p-cymene, carvacrol, linalool, and alpha-terpinene. These 5 monoterpenes also were assessed to determine their repellent activities to the mosquitoes. alpha-Terpinene had a potent repellent activity with a protection rate of 97% at a concentration of 0.05% topical treatment. Additionally, carvacrol and thymol showed an equivalent level of repellency. A spray-type solution containing 2% alpha-terpinene was tested for its repellent activity against Cx. pipiens. This solution showed stronger repellent activity than the currently used repellent N,N-diethyl-m methylbenzamide (deet). PMID- 12542195 TI - Colonization of Anopheles maculatus from Central Java, Indonesia. AB - The routine colonization of Anopheles maculatus, a reputed malaria vector from Central Java, is described. The strain is free mating and long lived in the laboratory. This species will readily bloodfeed on small rodents and artificial membrane systems. Either natural or controlled temperatures, humidity, and lighting provide acceptable conditions for continuous rearing. A simple larval diet incorporating a 10:4 powdered mixture of dried beef and rice hulls proved acceptable. Using a variety of simple tools and procedures, this colony strain appears readily adaptable to rearing under most laboratory conditions. This appears to be the first report of continuous colonization using a free-mating strain of An. maculatus. Using this simple, relatively inexpensive method of mass colonization adds to the short list of acceptable laboratory populations used in the routine production of human-infecting plasmodia. PMID- 12542194 TI - Evaluation of polymer-based granular formulations of Bacillus thuringiensis israelensis against larval Aedes aegypti in the laboratory. AB - A strategy to increase residual activity of Bacillus thuringiensis serovar. israelensis (Bti) extract through slow-release formulations and protection from solar radiation was studied. The median lethal concentration (LC50) and 90% lethal concentration (LC90) levels of laboratory-reared early 4th-stage larval Aedes aegypti after exposure to Bti extract were determined. Formulations with 4 polymers and 1 solar protectant were prepared, and their shelf lives were evaluated for year-long storage at 20-35 degrees C and 50-80% relative humidity. Also, the effect of ultraviolet light on unformulated (extract) and formulated Bti larvicidal activity persistence was determined. Laboratory bioassays were conducted with larval Ae. aegypti introduced into treated and control containers at 1, 2, 7, 14, 21, 28, 35, and 42 days after treatment, and larval mortalities were checked 24 h after introduction. Probit analysis of Bti extract showed LC50 and LC90 values of 0.016 and 0.051 mg/liter, respectively. The polymer-based Bti formulations showed no significant loss of insecticidal activity after 8 months of storage. Ultraviolet irradiation reduced activity of unformulated Bti extract after different exposure times, up to 40-46%, whereas Bti formulated with gelatin or acacia gum showed lower variation in larvicidal activity than formulations with sodium alginate and paraffin for protecting the activity of Bti toxin. Residual activity against 4th-stage Ae. aegypti in the laboratory for the formulation containing acacia gum at 10% (w/w) was 80% mortality at 14 days after treatment, whereas the Bti formulation containing gelatin (10%, w/w) caused 65% mortality. In addition, Bti formulations made with paraffin at 5% (w/w) sustained up to 60% mortality for 21 days. Unformulated Bti showed only 2.6% mortality, and a commercial preparation maintained 37% mortality, both at 14 days after treatment. PMID- 12542196 TI - Is it time to eliminate confusion regarding cervical lymph node levels according to the scheme originated at the Memorial Sloan-Kettering Cancer Center? PMID- 12542197 TI - Decrease in cochlear blood flow with infusion of nitric oxide synthase inhibitor and its recovery with L-arginine infusion: comparison with abdominal blood flow and auricular blood flow. AB - We observed changes in cochlear blood flow (CoBF), abdominal blood flow (AbBF) and auricular blood flow (AuBF) in rats after administration of the nitric oxide (NO) synthase inhibitor N-nitro-L-arginine-methyl ester (L-NAME). Ten min after i.v. infusion of L-NAME, L-arginine, a substrate of NO, was infused (100 mg/kg) intravenously. Using a laser Doppler flowmeter, changes in blood flow were recorded from the basal turn of the right cochlea, abdominal wall or right auricle, and systemic blood pressure (BP) was recorded simultaneously from the left femoral artery. As another parameter of vascular response, vascular conductance (VC) was calculated from blood flow/mean BP. I.v. infusion of L-NAME produced a dose-dependent depression of cochlear VC at concentrations of 0.2 ( 18.9%), 1 (-37.9%), 5 (-45.8%) and 10 mg/kg (-48.3%). Infusion of L-arginine partially reversed the decrease of CoBF caused by L-NAME. The group that received a 0.2 mg/kg infusion of L-NAME showed the largest degree of recovery (23.8%) with L-arginine, while the 10 mg/kg group showed the smallest degree of recovery (10.1%). AbBF also decreased after infusion of L-NAME (5 mg/kg) but to a lesser degree (-41.1% depression of VC) and with no significance compared to the decrease in CoBF (5 mg/kg L-NAME). Likewise, AuBF showed a decrease (-44.0% depression of VC) after infusion of L-NAME (10 mg/kg), the decrease being less than that of CoBF (10 mg/kg L-NAME). Recoveries from these decreased levels of VC in the AbBF and AuBF groups appeared to be smaller than those in the CoBF groups at the same dose of L-NAME (5.5% vs 17.3% in abdominal VC; 5.3% vs 10.1% in auricular VC). In a previous study comparing the CoBF changes caused by i.v. infusion and round window application of L-NAME, we proposed that i.v. infusion of L-NAME in rats primarily affects the precapillary arteriole of the spiral modiolar artery, which effectively regulates cochlear microcirculation as a resistance artery. Thus we assume that there may exist an active pathway of the NO/soluble guanylate cyclase/cyclic guanosine monophosphate system in the above vessels. With regard to the finding of a smaller recovery of VC with L-arginine in both the AbBF and AuBF groups, we consider that differences in L-arginine availability or uptake, and in the synthesis of NO, may exist between the cochlear and cutaneous vasculatures. PMID- 12542199 TI - Tympanic displacement analysis in healthy volunteers after indomethacin administration. AB - The aim of this study was to investigate whether a tympanic displacement analyser could detect decreases in cerebral blood flow and intracranial pressure after administration of indomethacin in healthy volunteers. In a double-blind crossover study involving 14 healthy volunteers all subjects first underwent a test-retest evaluation to investigate reproducibility followed by tests performed in sitting and supine positions to confirm intracranial-cochlear pressure transfer. In two further sessions tests were performed before and 90 min after subjects were blindly administered a suppository containing either 100 mg of indomethacin or placebo. It was found that tympanic membrane analysis performed 90 min after administration of such a suppository did not mirror the induced reduction in cerebral blood flow after administration of active drug. After administration of indomethacin eight of the subjects experienced discomfort and dizziness; after placebo none experienced subjective symptoms. After administration of indomethacin a statistically significant decrease in heart rate was demonstrated. The exponential form of the intracranial pressure-volume curve may explain why a decrease in intracranial pressure was not detected using the tympanic membrane displacement method, because the measurements were made in subjects with normal intracranial pressure. More significant findings may be found in patients with elevated intracranial pressure. PMID- 12542198 TI - Bumetanide-induced enlargement of the intercellular space in the stria vascularis requires an active Na+-K+-ATPase. AB - OBJECTIVE: Loop diuretics such as bumetanide and furosemide cause an acute enlargement of the intrastrial space of the stria vascularis, with an associated decline in the endocochlear DC potential (EP). The aim of this study was to determine the role played by the Na+-K+-ATPase in the bumetanide-induced enlargement of the intrastrial space, and to examine the importance of the balance between the activities of the Na+-K+-2Cl- cotransporter and the Na+-K+ ATPase to the physiological function of the stria vascularis. MATERIAL AND METHODS: Albino guinea pigs were used in experiments involving perilymphatic perfusion, EP measurement and electron microscopy. The effects of bumetanide on the stria vascularis were examined following inhibition of the Na+-K+-ATPase by ouabain. Ouabain was administered to the perfusate and, when the EP reached 0 mV, both ouabain and bumetanide were administered. RESULTS: Although there was no enlargement of the intrastrial space, vacuoles were apparent in marginal cells. The vacuolar change in marginal cells was similar to that caused by ouabain alone. CONCLUSION: This study indicates that the enlargement of the intrastrial space requires not only the blockade of the Na+-K+-2Cl- cotransporter but also normal activity of the Na+-K+-ATPase, and suggests that the bumetanide-induced enlargement of the intrastrial space resulted from the imbalance between the activities of the Na+-K+-2Cl- cotransporter and the Na+-K+-ATPase. PMID- 12542200 TI - Community-acquired methicillin-resistant Staphylococcus aureus infections in discharging ears. AB - OBJECTIVES: Although community-acquired methicillin-resistant Staphylococcus aureus (MRSA) infections have recently become an increasing problem in various fields of medicine, they have rarely been studied in the ear. The purpose of this study was to determine the prevalence of community-acquired MRSA infections in discharging ears and to compare the epidemiology of MRSA with that of other pathogens. MATERIAL AND METHODS: Between August 2000 and February 2002, 248 isolates recovered from 221 discharging ears of patients with 3 disease entities (chronic otitis media, acute otitis externa and granular myringitis) were prospectively analyzed using bacteriology. The isolates analyzed using bacteriology were divided into three groups: MRSA, methicillin-susceptible S. aureus (MSSA) and non-SA. RESULTS: S. aureus was found in 108 (43.5%) isolates and MRSA in 27. Therefore, the percentage of MRSA among the S. aureus isolates was 25.0% (27/108). The prevalence of community-acquired MRSA infections in discharging ears was 12.2% (27/221) and the MRSA strains were highly susceptible to vancomycin, teicoplanin, fusidic acid and minocycline. The MRSA-infected patients were older than those infected with other groups of pathogens. Also, MRSA infections appeared to be more common in chronic otitis media than in acute otitis externa or granular myringitis. CONCLUSIONS: Community-acquired MRSA represents an increasing problem. In this study, MRSA strains were highly susceptible to vancomycin, teicoplanin, fusidic acid and minocycline. PMID- 12542201 TI - 3D CT imaging method for measuring temporal bone aeration. AB - OBJECTIVE: 3D volume reconstruction of CT images can be used to measure temporal bone aeration. This study evaluates the technique with respect to reproducibility and acquisition parameters. MATERIAL AND METHODS: Helical CT images acquired from patients with radiographically normal temporal bones using standard clinical protocols were retrospectively analyzed. 3D image reconstruction was performed to measure the volume of air within the temporal bone. The appropriate threshold values for air were determined from reconstruction of a phantom with a known air volume imaged using the same clinical protocols. The appropriate air threshold values were applied to the clinical material. RESULTS: Air volume was measured according to an acquisition algorithm. The average volume in the temporal bone CT group was 5.56 ml, compared to 5.19 ml in the head CT group (p = 0.59). The correlation coefficient between examiners was > 0.92. There was a wide range of aeration volumes among individual ears (0.76-18.84 ml); however, paired temporal bones differed by an average of just 1.11 ml. CONCLUSIONS: The method of volume measurement from 3D reconstruction reported here is widely available, easy to perform and produces consistent results among examiners. Application of the technique to archival CT data is possible using corrections for air segmentation thresholds according to acquisition parameters. PMID- 12542202 TI - Osteoblast-like cell cultures from human stapes. AB - OBJECTIVE: In order to develop new middle ear prostheses for ossicular reconstruction it is important to study how the recipient middle ear tissues, especially the stapes footplate and superstructure, react to the implanted biomaterial. In this respect, animal studies and cell cultures using non-specific cells are of limited value. MATERIAL AND METHODS: The morphology and growth pattern of cells cultured from human stapes were studied. Cultured cells were examined for the presence of alkaline phosphatase and were processed for immunocytochemistry in order to detect the presence of osteocalcine. Fibroblast cultures served as controls. RESULTS: Cultured stapes cells proliferated in a polygonal-cubic shape and without any regular pattern in the culture. These cells were shown to contain alkaline phosphatase and osteocalcine. Cultured fascia fibroblasts proliferated in a spindle-shaped form and in a pattern resembling a shoal of fish. Cultured fibroblasts did not contain alkaline phosphatase or osteocalcine. CONCLUSIONS: It is possible to culture osteoblasts from human stapes. These cells can be characterized as osteoblast-like cells by means of their external shape and the presence of alkaline phosphatase and osteocalcine. Using these cultures, specific in vitro investigations concerning the interaction of biomaterials and middle ear ossicles could be performed. PMID- 12542203 TI - Factors affecting recovery after acoustic neuroma resection. AB - In order to determine which variables influence recovery from acoustic neuroma resection, acutely postoperative patients were divided into two groups and treated twice daily at bedside as inpatients at a tertiary care center. Treatment times were increased gradually from 5 to 30 min. Exercise subjects underwent passive and active head movements and walking with augmented head movements. Control subjects received attention without exercise. Pre- and post-tests included vertigo intensity and frequency, low-frequency vestibulo-ocular reflex (VOR), posturography and path integration. Post-tests were given at discharge and at approximately 4-week intervals for 3 months. Multilevel statistical analyses showed that neither group assignment nor age affected the outcome. All measures were abnormal at discharge but improved towards normal within 3 weeks. VOR on the unoperated side returned to normal values but on the lesioned side did not. Therefore, compensation is influenced by tumor size but not by age or early postoperative vestibular rehabilitation. Most compensation occurs within 3 weeks, probably due to central mechanisms, although some measures may not recover completely. PMID- 12542204 TI - Influence of human leukocyte antigen in the pathogenesis of Meniere's disease in the South Korean population. AB - In order to study the associations of Meniere's disease (MD) with human leukocyte antigen (HLA) alleles in the South Korean population, we examined the distribution of HLA class I and II alleles in 39 MD patients and 199 healthy controls. The frequencies of HLA-Cw*0303 (relative risk (RR) = 2.5; p < 0.02) and -Cw*0602 (RR = 3.7; p < 0.03) were significantly increased in patients with MD compared to controls. However, HLA-B44 (RR = 0.2, p < 0.004) and -Cw*0102 (RR = 0.3; p < 0.03) were significantly decreased in patients compared to controls. When the association between hearing level and the presence of HLA alleles was evaluated, the frequencies of HLA-B13 (RR = 7.4; p < 0.004), -B39 (RR = 45.7; p < 0.003), -Cw*0303 (RR = 4.5; p < 0.02) and -Cw*0602 (RR = 6.5; p < 0.02) were significantly increased and those of -B44 (RR = 0.1; p < 0.02) and -Cw*0102 (RR = 0.1; p < 0.03) were significantly decreased in MD patients with hearing losses (stages II-IV) compared to controls. The frequency of HLA-B13 was significantly higher in MD patients with hearing loss (stages II-IV) than in those with normal hearing (stage I). In terms of HLA-DRB1, -DQA1 and -DQB1 allele frequencies, a significantly higher frequency of DRB1*15 was seen in the patients (RR = 2.4; p < 0.03). These results suggest that some HLA alleles may be a useful genetic marker for predicting prognosis in South Korean patients with MD. PMID- 12542205 TI - Increased CD4+ T cells during acute attack of Meniere's disease. AB - OBJECTIVE: We aimed to investigate if there is an alteration in CD4 and CD8 T cell populations and in CD25 and CD49d activation antigens linked to these cells during an acute attack of Meniere's disease (MD). MATERIAL AND METHODS: Patients with MD who met the American Academy of Otolaryngology-Head and Neck Surgery criteria for the disease and healthy controls were enrolled in this study. Blood samples were collected during an acute attack of MD and when the disease was in its quiescent phase. RESULTS: 16 patients with MD and nine healthy controls were enrolled. The percentage of CD4 (T helper) cells was significantly increased during an acute attack of MD compared to that in healthy controls and in patients in the quiescent phase (71% vs 58% and 59%, respectively; p < 0.05). The increase in activation antigens was not statistically significant. CONCLUSION: To our knowledge this is the first demonstration of an increase in CD4 cells during an acute attack of MD. Further investigation is required to clarify the role of T cells in the pathogenesis of the disease. PMID- 12542206 TI - The paranasal sinuses as reservoirs for nitric oxide. AB - OBJECTIVE: Nitric oxide (NO) is an important mediator and inflammatory marker in human upper airways. Enzymes responsible for NO production have been demonstrated both in the nose and in the paranasal sinuses, but NO levels in the sinuses are reported to be several times higher than those in the nose. It has been postulated that the paranasal sinuses may be the primary sites for NO production in the upper respiratory tract. The present study was designed to compare the NO levels sampled from the nose with those found in the paranasal sinuses. MATERIAL AND METHODS: NO levels in the maxillary sinus and nose were determined using a continuous chemiluminescence measuring technique in seven healthy volunteers. RESULTS: When NO was sampled, via a drainage tube inserted into the maxillary sinus, a transient peak in NO level was recorded. The maximal NO level (5,761 +/- 1,513 ppb; n = 7) was reached within 10 s and was followed by the establishment of a lower steady-state level (304 +/- 51 ppb). When NO was continuously sampled from the nose a steady-state level, similar to that found in the sinus, was immediately established (313 +/- 52 ppb). CONCLUSION: The data presented confirm previous findings of extremely high NO levels in the paranasal sinuses and suggest that these cavities may also function as reservoirs for NO. PMID- 12542207 TI - Expression of mRNA transcripts of the Na+/H+ and Cl-/HCO3- exchanger isoforms in human nasal mucosa. AB - Electrolyte transport by nasal epithelia has been suggested to be important for controlling the quantity and composition of the nasal fluid and may play an important role in the development of nasal polyps. One of a number of mechanisms involving translocation of Na+ and Cl- across cell membranes includes electroneutral processes, such as Na+/H+ exchange (NHE) and Cl-/HCO3- exchange (AE). The present study evaluated the presence of mRNAs for various members of the human NHE and AE gene families in human inferior turbinate mucosa and nasal polyp using reverse transcriptase polymerase chain reaction and in situ hybridization. The mRNA for NHE1 was detected in human turbinate mucosa and nasal polyp while the mRNAs for NHE2 and NHE3 could not be detected in any of the samples examined. Of the AE isoforms, AE2 mRNA was expressed in inferior turbinate mucosa but not in nasal polyp. In situ hybridization revealed that NHE1 mRNA in the turbinate mucosa and nasal polyp was localized in the epithelial layer and submucosal glands. AE2 mRNA was also expressed in the epithelial layer and submucosal glands of inferior turbinate mucosa. Taken together, these results indicate that the expression of AE2 mRNA is altered in nasal polyp compared with inferior turbinate mucosa, suggesting that the altered expression of these genes in nasal polyp may cause impaired electrolyte and water transport across the epithelial cells. PMID- 12542208 TI - Presence of platelet-activating factor in nasal polyps and eosinophils. AB - Platelet-activating factor (PAF) has been reported to play a role in allergy and inflammatory reactions but its role in the pathogenesis of nasal polyps remains unclear. In this study, we examined both PAF and peptide leukotrienes (peptLTs) in individual preparations from nasal polyps. The amounts of PAF were much greater than those of peptLTs in all preparations. Nasal polyps were divided into two groups according to the severity of eosinophil infiltration: a severe group (eosinophil count > or = 50/mm2) and a mild group (eosinophil count < 50/mm2). The amounts of PAF in the nasal polyps were significantly higher in the severe group than in the mild group (p < 0.01). PAF activity correlated with tissue eosinophilia and polyps obtained from patients with aspirin-sensitive asthma contained relatively large amounts of PAF, with enriched infiltration of eosinophils. PMID- 12542209 TI - The quinoxaline derivative caroverine in the treatment of sensorineural smell disorders: a proof-of-concept study. AB - The treatment of non-conductive olfactory disorders is to a large extent an unsolved problem. This proof-of-concept study focused on possible effects of the N-methyl-D-aspartate (NMDA) antagonist caroverine. Potential mechanisms for the hypothesized effect included reduced feedback inhibition in the olfactory bulb as a consequence of NMDA antagonistic actions and antagonism of an excitotoxic action of glutamate. A total of 77 consecutive patients with non-conductive olfactory disorders were included in the study. Fifty-one patients received caroverine for 4 weeks (120 mg/day); 26 controls matched for age, gender and duration of olfactory loss were treated with zinc sulfate for the same length of time (400 mg/day). Olfactory sensitivity was evaluated before and after treatment. Testing included assessment of n-butanol odor threshold and odor identification. When compared to baseline, treatment with caroverine improved both odor thresholds (p = 0.005) and odor identification (p = 0.042) in anosmic patients. In hyposmic patients it significantly improved odor identification ability (p = 0.041). In contrast, zinc sulfate had no significant effect on olfactory function. These results indicate that caroverine appears to be effective for the treatment of non-conductive smell disorders. PMID- 12542210 TI - Cricoid area of the larynx: its physiological and pathological significance. AB - The three-dimensional distribution of the cricoid area was investigated using computer graphics and its histological structure and pathology were studied using whole-organ serial sections. A total of 26 adult human larynges were examined. The findings were as follows: 1. Cricoid areas were located along the superior portion of the cricoid arch on both sides. 2. The cricoid area was surrounded by the perichondrium of the cricoid cartilage, the conus elasticus and the fibrous layer of the subglottic mucosa. 3. The cricoid area was a loose areolar area, mainly composed of adipose tissue and loose elastic and collagenous fibers. 4. Many vessels were present in the cricoid area and a superficial branch of the cricothyroid artery ran through it. 5. Vessels in the cricoid area penetrated the anteroinferior portion of the conus elasticus and extended into the prelaryngeal region. 6. In larynges with laryngeal carcinoma, cancer invasion into the cricoid area and intravascular tumor invasion facilitated metastasis to the prelaryngeal, pretracheal and/or paratracheal regions and stomal recurrence. Cricoid areas were related to the growth pattern of laryngeal cancer. PMID- 12542211 TI - Sinonasal adenocarcinoma: clinical study of nine cases in Taiwan. AB - OBJECTIVE: Sinonasal adenocarcinoma is an uncommon disease in Taiwan. This paper presents details of the management and outcome of the disease. MATERIAL AND METHODS: The medical records of patients with sinonasal adenocarcinoma who presented to the Department of Otolaryngology, National Taiwan University Hospital between 1983 and 2002 were retrospectively reviewed. Tumors were staged based on the American Joint Committee of Cancer staging criteria. The series included 5 males and 4 females (mean age 51 years; range 28-75 years). RESULTS: No association between sinonasal adenocarcinoma and occupations such as furniture making, wood working or textile working was found. Epistaxis and nasal obstruction were the most common symptoms. Metastasis to bone or liver was found initially at diagnosis in two cases. Treatment was given according to the tumor stage, the clinical condition of the patient and the patient's wishes: two received combined treatment comprising wide surgical resection and radiotherapy, two received radiotherapy only, one received wide surgical resection only and four received no treatment. At a mean follow-up of 30 months (range 2-68 months), 2 patients were free from the disease and 1 was alive with the disease. Six cases died of the disease. Advanced-stage disease was associated with a poor prognosis. Early-stage disease, a well-differentiated histologic type and surgical resection combined with radiotherapy were associated with a favorable result. CONCLUSION: Sinonasal adenocarcinoma has a low incidence in the general population in Taiwan. However, prognosis is poor owing to delayed diagnosis and the consequent advanced stage of the disease. PMID- 12542212 TI - Personality traits in head and neck squamous cell carcinoma patients in relation to the disease state, disease extent and prognosis. AB - OBJECTIVE: We aimed to study whether personality scores, as measured using the Eysenck Personality Inventory (EPI), are associated with the risk and prognosis of head and neck squamous cell carcinoma (HNSCC). MATERIAL AND METHODS: A total of 78 male patients with newly diagnosed HNSCC and 61 male patients with benign HN disease completed the EPI, which assesses the following: neuroticism, extraversion and lie score. The TNM stage, prognosis, diagnostic delay, level of education and smoking and alcohol consumption histories of the patients were also recorded. Patients with cachexia and those aged > 80 years were excluded. RESULTS: The cancer patients had higher neuroticism scores (10.7 +/- 0.5 vs 8.3 +/- 0.6; p < 0.01) than the control patients. This association was shown to be secondary to the fact that neuroticism is associated with increased alcohol consumption. The personality trait scores were not associated with the length of diagnostic delay. It was also suggested that a low lie score predicted disease specific death in the HNSCC patients (p = 0.02). Total survival also seemed to be predictable (p < 0.05). CONCLUSION: Neuroticism is probably associated with a risk of HNSCC as a result of increased alcohol consumption. If the prognostic results can be replicated, a potentially important association between this personality trait and the prognosis of HNSCC is suggested. PMID- 12542213 TI - Current management of cutaneous malignant melanoma of the head and neck. PMID- 12542214 TI - The syndrome of inappropriate antidiuretic hormone secretion after bilateral radical neck dissections. AB - OBJECTIVE: It is commonly believed that synchronous, bilateral radical neck dissection causes the syndrome of inappropriate secretion of antidiuretic hormone (SIADH), presumably as a result of increased intracranial pressure. This belief is based mainly on the results of an experimental study published in 1978 in which occlusion of the superior vena cava in dogs resulted in increased intracranial pressure and SIADH. The purpose of the present study was to test this assumption in an animal model that more closely resembles the clinical condition of bilateral radical neck dissection, in an attempt to discover possible surgical methods for preventing SIADH. MATERIALS AND METHODS: The experiments were carried out in six adult mongrel dogs. In two animals the internal and external jugular veins were ligated on both sides of the neck, high in the neck and at the thoracic inlet, respectively. In four animals, a formal radical neck dissection was performed on both sides of the neck. Urine and serum osmolarity and serum electrolyte levels were measured prior to surgery and at 2.5, 24, 48 and 120 h after surgery, in order to monitor the development of SIADH. RESULTS: No significant changes in serum osmolarity, urine osmolarity or serum sodium levels were observed in any of the animals studied. CONCLUSIONS: Bilateral synchronous jugular vein ligation and bilateral radical neck dissections did not result in SIADH in a dog model. As these results contradict a commonly held belief in clinical practice, a prospective evaluation of the physiologic changes after bilateral radical neck dissections is warranted. PMID- 12542215 TI - Bronchopneumonia and oral health in hospitalized older patients. A pilot study. AB - AIMS: To correlate microbial findings obtained by bronchoalveolar lavage in pneumonia patients with the clinical situation of the oral cavity. METHOD: Quantitative aerobic and anaerobic cultures were carried out in 150 ml samples of bronchoalveolar lavage (BAL) obtained by means of an endoscope (Video Endoscope Pentax) inserted per os in the infected bronchus. MATERIAL: Twenty consecutive patients with a tentative clinical diagnosis of bronchopneumonia in whom BAL was carried out for diagnostic purposes. A clinical evaluation of the oral health status (oral hygiene, caries, periodontal diseases) was subsequently carried out. RESULTS: In seven edentulous subjects wearing complete dentures the culture of anaerobic microorganisms was negative or yielding less than 100 cfu/ml BAL. Two patients yielded high counts of S. aureus and one high counts of P. aeruginosa. In the 13 subjects with natural teeth left one showed high counts of Veillonella spp. (anaerobic) + P. aeruginosa, one high counts of Veillonella spp. + S. aureus, one high counts of P. aeruginosa + S. aureus and one high counts of E. coli. These four subjects showed poor oral hygiene, periodontal pockets and a BAL microflora consistent with periodontal pathology. CONCLUSION: The results of this pilot study suggest that microorganisms of denture plaque or associated with periodontal diseases may give rise to aspiration pneumonia in susceptible individuals. PMID- 12542216 TI - Prosthetic crowns and other clinical risk indicators of caries among old-old Swedish adults: findings from the KEOHS Project. Kungsholmen Elders Oral Health Study. AB - OBJECTIVES: The Kungsholmen Elders Oral Health Study (KEOHS) evaluated the oral health status of generally healthy, community-dwelling persons over the age of 80 living in Kungsholmen, Sweden. This paper explored possible clinical risk indicators of coronal and root caries among the KEOHS subjects. DESIGN: In this cross-sectional study, dentate KEOHS subjects received a caries assessment using defined visual, tactile criteria. SETTING: Examinations were carried out in two local clinics by standardized examiners. SUBJECTS: One hundred twenty-nine dentate persons were examined. MAIN OUTCOME MEASURES: The examination identified decayed and filled surfaces, prosthetic crowns, and missing teeth. RESULTS: More root than coronal surfaces had untreated decay, and secondary root caries contributed the greatest number of decayed surfaces. Ninety percent of the examined dentate subjects had at least one prosthetic crown. Root surfaces exposed to crown margins were more likely to have caries than root surfaces not so exposed, particularly among women. The presence of untreated coronal caries (yes/no) was positively associated with having untreated root caries and an intermediate number (14-20) of teeth, but inversely associated with having 4+ prosthetic crowns. Active root caries (yes/no) was positively associated with having untreated coronal caries, 14-20 teeth, and 4+ prosthetic crowns. Nearly 20% of identified root lesions were present at or below the gingival margin, and most (88%) were secondary caries associated with crown margins (65%) or other restorations (23%). CONCLUSIONS: Our findings suggest that some dental characteristics, including the presence of prosthetic crowns, are risk indicators for the presence of untreated coronal and root caries. PMID- 12542217 TI - Caries incidence and increments in community-living older adults with and without dementia. AB - OBJECTIVES: The Oral Health of Older Adults with Dementia was instigated in the late 1990s to quantify and compare coronal and root caries incidence and increments in community-living older Australians with and without dementia. METHODS: A longitudinal design was used to conduct dental inspections at baseline and one-year, for two groups of randomly selected community-living older adults- one group of 116 people with dementia and a comparison group of 116 people without dementia. RESULTS: At one-year there were 103 dementia and 113 (112 dentate) non-dementia participants. Coronal and root surface caries incidence was higher for dementia participants (p < 0.05). Dementia participants had higher coronal and root caries adjusted caries increments (ADJCI) (p < 0.01). Both coronal and root ADJCI were evident in half of dementia participants, compared with one-quarter of non-dementia participants. Dementia participants with higher coronal ADJCI were those who had visited the dentist since baseline, who were taking neuroleptics with high anticholinergic adverse effects, and whose carer had high carer burden score (p < 0.01). Dementia participants with higher root ADJCI were those needing assistance with oral hygiene care and whose carers had difficulties with oral hygiene care (p < 0.05). Baseline characteristics predictive in linear regression for: (1) coronal caries increments among all participants were--dementia participants, those with cognitive testing scores indicative of moderate-severe dementia, those with private health insurance; (2) root caries increments among all participants were--dementia participants, and those who had > or = 1 decayed/filled root surface at baseline. Among dementia participants, being male was the baseline characteristic predictive in logistic regression for coronal caries increments, and having > or = 1 decayed coronal surface was the baseline characteristic predictive for root caries increments. CONCLUSIONS: Coronal and root caries incidence and increments were significantly higher in the community-living older adults with dementia over the one-year follow-up period. Dementia participants had high levels of coronal and root caries increments; characteristics related to high caries increments included sex (males), dementia severity (moderate-severe), high carer burden, oral hygiene care difficulties, use of neuroleptic medication (with high anticholinergic adverse effects) and previous experience of caries. PMID- 12542218 TI - Masticatory ability in 80-year-old subjects and its relation to intake of energy, nutrients and food items. AB - OBJECTIVE: The aim of the present study was to analyse the relationship between masticatory ability (self-assessed masticatory ability and bite force) and intake of energy, nutrients and food items in a population sample of elderly subjects. DESIGN AND SUBJECTS: From a population sample of 80-year-old people, 160 individuals (74 men and 86 women) took part in an odontological study. MAIN OUTCOME MEASURES: A dental examination including bite force recording, a questionnaire focusing on self-assessed masticatory ability, and a dietary interview. SETTING: Department of Geriatric Medicine, Goteborg University, Sweden. RESULTS: The dental status among the participants varied much (from edentulous in both jaws--22%--to more than 20 natural teeth--30%). The mean maximum bite force was higher in men (165 N) than in women (105 N). Bite force was significantly correlated to the Eichner index and to the number of teeth. One third of the subjects reported no masticatory problem, whereas 18% identified > or = 3 such problems. The intake of energy and nutrients varied much but the means were well above recommended values. The correlations between dental status and bite force on one side and dietary intake on the other side were in general weak and most often statistically non-significant. Impaired general health and reduced dentition were both associated with more masticatory problems. CONCLUSION: The examined sample of 80-year-old subjects had a great variation in dental status, bite force and self-assessed masticatory ability, but these factors had only a minor influence on dietary selection and intake, which on average were well above recommended values. PMID- 12542219 TI - Testing a model of the relationship between gender, ethnicity, clinical status and impact in older adults from minority ethnic groups. AB - OBJECTIVE: To identify the relationship between indicators of self-assessed symptom status, the reported impact of oral conditions and clinical indices, and the extent to which this relationship was moderated by gender and ethnicity. DESIGN: Secondary analysis of data from an oral health survey of minority ethnic groups. PARTICIPANTS: Purposive sample of 376 individuals from minority ethnic groups in the United Kingdom recruited through community groups. MEASURES: Numbers of decayed, missing and filled teeth. Measures of self assessed symptoms, and impact upon quality of life. RESULTS: Impact of oral conditions upon lifestyle was predicted by the number of missing teeth, the presence of pain on eating certain foods and the presence of toothache in the previous four weeks. Social variables (gender and ethnicity) did not predict impact either singly or through interaction with symptoms. CONCLUSIONS: The findings support a linear model of the relationship between the experience of oro-facial symptoms and impact on everyday life amongst older adults. PMID- 12542220 TI - Oral health behind bars: a study of oral disease and its impact on the life quality of an older prison population. AB - OBJECTIVE: The principal aims of this study were to report on the prevalence of oral disease and its impact on the life quality of older (aged 60 and above) prisoners at Hong Kong SAR's elderly detention centre, China. DESIGN: A cross sectional epidemiological survey involving clinical oral examinations and face-to face interviews with 64 older prisoners at the centre. Clinical oral examinations were carried out following WHO criteria. The impact of oral health on life quality was assessed using the 14-item Oral Health Impact Profile measure, OHIP 14. RESULTS: The prevalence of oral disease was high, the mean DMFT was 22.5 (SD 10.6) and 40% (18) of the dentate prisoners had shallow or deep periodontal pockets. Most (75%, 48) were in need of prosthetic treatment. Over half the prisoners experienced one or more oral health impacts on life quality during the previous year. This was associated with socio-demographic factors: employment status prior to incarceration (P < 0.01), oral health behaviour: smoking habits (P < 0.01) and clinical oral health status: DMFT (P < 0.05) and prosthetic need (P < 0.05). CONCLUSION: Among inmates at Hong Kong's elderly detention centre, the prevalence of oral disease was high and the impact of oral health on their life quality was substantial. The impact of oral health on the life quality of the older prisoners was associated with prisoners socio-demographic background, oral health behaviour and oral health status. PMID- 12542221 TI - Utilization of dental health services by community-dwelling older adults in Japan who attended a weekly educational programme. AB - OBJECTIVES: The purpose of this study was to describe dental health services utilization and identify factors which influenced a group of independently living elderly persons in an urban area of Japan. SUBJECTS: The study sample consisted of 2,990 participants, 83% of the total sample of the Senior Citizen's College, who were 60 years and over. MEASUREMENTS: Their dental utilization and satisfaction with dental treatment were measured by a questionnaire from 1995 to 1998. RESULTS: The mean age of the subjects was 66.5 years and 52% were male. Sixty percent of the subjects had visited a dentist within the previous year, and 33% of them had received a regular oral health check-up. The majority of the subjects (63%) were satisfied with their dental treatment. A multiple stepwise logistic regression analysis showed that dental services utilization had a significant positive association with presence of teeth (p < 0.001), being male (p < 0.01) and satisfaction with financial status (p < 0.05). A regular oral health check-ups had a significant association with presence of teeth (p < 0.001), satisfaction with financial status (p < 0.05) and aging (p = 0.001). CONCLUSIONS: Dental health services utilization was related to the presence of teeth and financial status, rather than age or medical conditions, among independently living elderly persons in an urban area of Japan. PMID- 12542222 TI - Difficulties in dental prescribing of saliva substitutes for xerostomia. AB - The value of salivary substitutes for many elderly patients is indicated by a simple web search which found 2,470 references, with references to prescribing across the globe. To contain the public expense, prescribing medicaments within the National Health Service in England is limited to approved lists. The only salivary substitute currently prescribable by dentists has not been produced for 18 months. It is argued that: the prescription of this symptomatic therapy should be based on potential benefit for the xerostomic patient and not, as now, primarily on the pathological aetiology of the oral dryness. Secondly, that the professional caring for that oral problem--the dentist, is best placed to manage the prescribing as part of the overall management of the patient, independent of any reference to the patients physician that may be appropriate as part of the overall management of the patient. PMID- 12542223 TI - Facilitated CA1 hippocampal synaptic plasticity in dystrophin-deficient mice: role for GABAA receptors? AB - Duchenne muscular dystrophy (DMD) is associated with cognitive deficits that may result from a deficiency in the brain isoform of the cytoskeletal membrane associated protein, dystrophin. CA1 hippocampal short-term potentiation (STP) of synaptic transmission is increased in dystrophin-deficient mdx mice, which has been attributed to a facilitated activation of NMDA receptors. In this study, extracellular recordings in the hippocampal slice preparation were used first to determine the consequences of this alteration on short-term depression (STD). STD induction was facilitated in mdx as compared with wild-type mice in a control medium. Because brain dystrophin deficiency results in a decreased number of gamma-aminobutyric acid A (GABAA)-receptor clusters, we tested the hypothesis that neuronal disinhibition contributes to the enhanced synaptic plasticity in mdx mice. We found that the GABAA receptor antagonist, bicuculline, increased basal neurotransmission in wild-type, but not in mdx mice and prevented the enhanced STP and STD in the CA1 area of slices from mdx mice. The possibility that altered GABA mechanisms underlie the facilitation of NMDA receptor-dependent synaptic plasticity in mdx mice is discussed. PMID- 12542224 TI - Differential contributions of the parahippocampal place area and the anterior hippocampus to human memory for scenes. AB - Past neuroimaging research has identified a parahippocampal place area (PPA) in the posterior medial temporal lobe (MTL), which responds preferentially to visual scenes and plays a role in episodic memory for this class of stimuli. In the present positron emission tomography study, we examined to what extent the functional characteristics of the PPA resemble those of other, more anterior MTL regions across various learning and recognition-memory tasks. We also determined whether the involvement of the PPA in recognition of previously studied scenes is specific to a particular type of scene information. We found that, like the PPA, anterior hippocampal regions showed a novelty response (higher activation for novel than repeated scenes) and a stimulus-related response (higher activation for scenes than objects) during learning, indicating that MTL structures other than the PPA contribute to the encoding of novel stimulus relationships in scenes. However, these anterior hippocampal regions showed no involvement during recognition of either spatial or nonspatial information contained in scenes. The PPA, by contrast, was consistently involved in recognition of all types of scene details, presumably through interactions with co-activated parietal and occipitotemporal cortices. We suggest that MTL contributions from the PPA are sufficient to support recognition of scenes when the task can be based on a perceptually based familiarity process. PMID- 12542225 TI - Single neuron burst firing in the human hippocampus during sleep. AB - Although there are numerous non-primate studies of the single neuron correlates of sleep-related hippocampal EEG patterns, very limited hippocampal neuronal data are available for correlation with human sleep. We recorded human hippocampal single neuron activity in subjects implanted with depth electrodes required for medical diagnosis and quantitatively evaluated discharge activity from each neuron during episodes of wakefulness (Aw), combined stage 3 and 4 slow-wave sleep (SWS), and rapid eye movement (REM) sleep. The mean firing rate of the population of single neurons was significantly higher during SWS and Aw compared with REM sleep (p = 0.002; p < 0.0001). In addition, burst firing was significantly greater during SWS compared with Aw (p = 0.001) and REM sleep (p < 0.0001). The synchronized state of SWS and associated high-frequency burst discharge found in human hippocampus may subserve functions similar to those reported in non-primate hippocampus that require burst firing to induce synaptic modifications in hippocampal circuitry and in hippocampal projections to neocortical targets that participate in memory consolidation. PMID- 12542226 TI - Projections from the posterior cortical nucleus of the amygdala to the hippocampal formation and parahippocampal region in rat. AB - The posterior cortical nucleus of the amygdala is involved in the processing of pheromonal information and presumably participates in ingestive, defensive, and reproductive behaviors as a part of the vomeronasal amygdala. Recent studies suggest that the posterior cortical nucleus might also modulate memory processing via its connections to the medial temporal lobe memory system. To investigate the projections from the posterior cortical nucleus to the hippocampal formation and the parahippocampal region, as well as the intra-amygdaloid connectivity in detail, we injected the anterograde tracer phaseolus vulgaris-leucoagglutinin into different rostrocaudal levels of the posterior cortical nucleus. Within the hippocampal formation, the stratum lacunosum-moleculare of the temporal CA1 subfield and the adjacent molecular layer of the proximal temporal subiculum received a moderate projection. Within the parahippocampal region, the ventral intermediate, dorsal intermediate, and medial subfields of the entorhinal cortex received light to moderate projections. Most of the labeled terminals were in layers I, II, and III. In the ventral intermediate subfield, layers V and VI were also moderately innervated. Layers I and II of the parasubiculum received a light projection. There were no projections to the presubiculum or to the perirhinal and postrhinal cortices. The heaviest intranuclear projection was directed to the deep part of layer I and to layer II of the posterior cortical nucleus. There were moderate-to-heavy intra-amygdaloid projections terminating in the bed nucleus of the accessory olfactory tract, the central division of the medial nucleus, and the sulcal division of the periamygdaloid cortex. Our data suggest that via these topographically organized projections, pheromonal information processed within the posterior cortical nucleus can influence memory formation in the hippocampal and parahippocampal areas. Also, these pathways provide routes through which seizure activity can spread from the epileptic amygdala to the surrounding region of the temporal lobe. PMID- 12542227 TI - Coexistence of serotonin 3 (5-HT3) and CB1 cannabinoid receptors in interneurons of hippocampus and dentate gyrus. AB - Using in situ hybridization histochemistry, a high degree of coexpression of the functional 5-HT3A subunit of the 5-HT3 receptor and the central CB1 cannabinoid receptor was detected in all subfields of the hippocampus and subgranular layer of the dentate gyrus (DG). Semi-quantitative analysis demonstrated that, depending on the hippocampal layer, 72-88% of CB1-expressing interneurons coexpress the 5-HT3A subunit. Within the DG, 5-HT3A/CB1 double-labeled neurons were confined to the subgranular layer, where close to 80% of all CB1-expressing basket neurons were found to contain 5-HT3A subunit transcripts. These results provide the first evidence indicating that the only ion channel receptor for serotonin and central CB1 cannabinoid receptor coexist in neurons containing the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). These findings suggest possible interactions between the cannabinoid and serotonergic systems at the level of GABA neurotransmission. However, activation of 5-HT3- or CB1 receptors are likely to have opposing regulatory effects on GABA neurotransmission, as 5-HT3 receptor activation by serotonin results in the release of GABA, while CB1 activation by cannabinoids results in inhibition of GABA release. PMID- 12542228 TI - Trans-synaptic increase of hypoxic tolerance in hippocampus upon physical challenge with two-photon microscopy. AB - Neuronal hypoxic tolerance is modulated by preceding challenges. We investigated hypoxic tolerance in CA1 pyramidal cells of murine hippocampal slices upon preceding physical challenge with two-photon illumination in close spatial proximity to the recorded area, at distant presynaptic neurons, or preceding chemical treatment with acetylsalicylic acid while zinc fluorescence was assessed with fluorescence measurement upon staining with N-(6-methoxy-8-quinolyl)-para toluenesulfonamide (TSQ). Posthypoxic recovery (15 min hypoxia, 45 min recovery) of CA1 population spike amplitude (PSAP) upon stimulation of Schaffer collaterals in hippocampal region CA3 was 20 +/- 38% (mean +/- SD; n = 15) in control slices. At the end of hypoxia, zinc fluorescence increased to 120 +/- 16% (P < 0.05 to control) in slices that later recovered and 141 +/- 20% in slices that did not recover (P < 0.01 to control; P < 0.05 compared with returns). Multi-photon illumination alone was an appropriate physical challenge to improve hypoxic tolerance, even trans-synaptically. Depending on the number of illuminations posthypoxic PSAP increased up to 84 +/- 25% (P < 0.01 to control) upon illumination of hippocampal region CA1 and 85 +/- 28% (P < 0.01 to control) upon illumination of CA3. With the latter treatment, zinc fluorescence in CA1 increased to 126 +/- 20% before hypoxia (P < 0.05 to control), and no further zinc increase was observed upon subsequent hypoxia. Similar results were obtained upon chemical preconditioning with acetylsalicylate. We conclude that observation of live specimen with multi-photon imaging alters the physiology of neuronal cell ensembles, including hypoxic tolerance, even trans-synaptically at long distances from the imaged area. This is mediated in part through endogenous modulation by zinc. Mild zinc increase improves hypoxic tolerance while pronounced increase predicts neuronal cell death. PMID- 12542229 TI - Posttraining but not pretraining lesions of the hippocampus interfere with feature-negative discrimination of fear-potentiated startle. AB - Previous studies have suggested that the hippocampus may play an important role in some forms of inhibitory learning. The goal of the present study was to assess whether the hippocampus is also important for inhibition of fear acquired after serial feature-negative discrimination training. Rats were given aspiration lesions of the hippocampus either before or after training in which a target light was paired with shock when presented alone, but not paired with shock when presented in serial compound with a noise feature (light+/noise-->light-). Conditioned fear to the target stimulus and feature-target compound were measured with fear-potentiated startle. Pre-training lesion of the hippocampus did not disrupt feature-negative discrimination performance relative to sham-operated and cortical lesioned controls. In contrast, hippocampal lesions performed after training severely disrupted performance. Specifically, rats with hippocampal lesions failed to inhibit fear when the noise feature was presented in compound with the target. However, these rats could reacquire the feature-negative discrimination. These observations suggest that the hippocampus may normally be involved in retention or retrieval of serial feature-negative discrimination; however, in its absence feature-negative discrimination can still be acquired. PMID- 12542230 TI - Alterations in the balance of protein kinase and phosphatase activities and age related impairments of synaptic transmission and long-term potentiation. AB - Aging is associated with an impaired ability to maintain long-term potentiation (LTP), but the underlying cause of the impairment remains unclear. To gain a better understanding of the cellular and molecular mechanisms responsible for this impairment, the synaptic transmission and plasticity were studied in the CA1 region of hippocampal slices from adult (6-8 months) and poor-memory (PM)-aged (23-24 months) rats. The one-way inhibitory avoidance learning task was used as the behavioral paradigm to screen PM-aged rats. With intracellular recordings, CA1 neurons of PM-aged rats exhibited a more hyperpolarized resting membrane potential, reduced input resistance, and increased amplitude of afterhyperpolarization and spike threshold, compared with those in adult rats. Although a reduction in the size of excitatory synaptic response was observed in PM-aged rats, no obvious differences were found between adult and PM-aged rats in the pharmacological properties of excitatory synaptic response, paired-pulse facilitation, or frequency-dependent facilitation, which was tested with trains of 10 pulses at 1, 5, and 10 Hz. Slices from the PM-aged rats displayed significantly reduced early-phase long-term potentiation (E-LTP) and late-phase LTP (L-LTP), and the entire frequency-response curve of LTP and LTD is modified to favor LTD induction. The susceptibility of time-dependent reversal of LTP by low-frequency afferent stimulation was also facilitated in PM-aged rats. Bath application of the protein phosphatase inhibitor, calyculin A, enhanced synaptic response in slices from PM-aged, but not adult, rats. In contrast, application of the cAMP-dependent protein kinase inhibitors, Rp-8-CPT-cAMPS and KT5720, induced a decrease in synaptic transmission only in slices from the adult rats. Furthermore, the selective beta-adrenergic receptor agonist, isoproterenol, and pertussis toxin-sensitive G-protein inhibitor, N-ethylmaleimide, effectively restored the deficit in E-LTP and L-LTP of PM-aged rats. These results demonstrate that age-related impairments of synaptic transmission and LTP may result from alterations in the balance of protein kinase/phosphatase activities. PMID- 12542231 TI - Deterioration in learning and memory of inferential tasks for evaluation of transitivity and symmetry in aged SAMP8 mice. AB - This study examined age-dependent deficits in the learning and memory of inferential tasks, using an established senescence-accelerated mouse model in age related brain dysfunction (SAMP8) and its genetically related inbred strain (SAMR1). The mice learned two sets of nonspatial odor-odor pairs by association learning successively (i.e., A-->B, X-->Y, then B-->C, Y-->Z). They were tested in transitive inference (i.e., A-->C, X-->Z) and symmetrical inference (i.e., C- >B, Z-->Y). In the probe test of A-->C, X-->Z transitive inference, 1-month-old SAMP8 and control SAMR1 at the same age significantly chose the alternative based on transitive inference, but 4- and 7-month-old SAMP8 performed at a random chance level, in comparison with unambiguous inference by control SAMR1 at the same ages. During the test of C-->B, Z-->Y symmetrical inference, SAMP8 at 1 month of age made errors as frequently as control SAMR1 at the same age, but SAMP8 at 4 and 7 months of age made more errors than SAMR1 at the same ages. At 4 and 7 months of age, SAMP8 made more errors than 1-month-old SAMP8. Control SAMR1 did not show such an age-related deficient. These results indicate that SAMP8 mice have age-related learning and memory deficits in the ability to perform inferential tasks. Age-related hippocampal dysfunction is suggested to be the cause of these age-related deficits in old SAMP8 mice during the performance of inferential tasks mediated by declarative memory. PMID- 12542232 TI - Human hippocampus and viewpoint dependence in spatial memory. AB - Virtual reality was used to sequentially present objects within a town square and to test recognition of object locations from the same viewpoint as presentation, or from a shifted viewpoint. A developmental amnesic case with focal bilateral hippocampal pathology showed a massive additional impairment when tested from the shifted viewpoint compared with a mild, list length-dependent, impairment when tested from the same viewpoint. While the same-view condition could be solved by visual pattern matching, the shifted-view condition requires a viewpoint independent representation or an equivalent mechanism for translating or rotating viewpoints in memory. The latter mechanism was indicated by control subjects' response latencies in the shifted-view condition, although the amnesic case is not impaired in tests of mental rotation of single objects. These results show that the human hippocampus supports viewpoint independence in spatial memory, and suggest that it does so by providing a mechanism for viewpoint manipulation in memory. In addition, they suggest an extremely sensitive test for human hippocampal damage, and hint at the nature of the hippocampal role in episodic recollection. PMID- 12542234 TI - User-oriented views in health care information systems. AB - In this paper, we present the methodology we adopted in designing and developing an object-oriented database system for the management of medical records. The designed system provides technical solutions to important requirements of most clinical information systems, such as 1) the support of tools to create and manage views on data and view schemas, offering to different users specific perspectives on data tailored to their needs; 2) the capability to handle in a suitable way the temporal aspects related to clinical information; and 3) the effective integration of multimedia data. Remote data access for authorized users is also considered. As clinical application, we describe here the prototype of a user-oriented clinical information system for the archiving and the management of multimedia and temporally oriented clinical data related to percutaneous transluminal coronary angioplasty (PTCA) patients. Suitable view schemas for various user roles (cath-lab physician, ward nurse, general practitioner) have been modeled and implemented on the basis of a detailed analysis of the considered clinical environment, carried out by an object-oriented approach. PMID- 12542233 TI - Transcriptional profiling reveals regulated genes in the hippocampus during memory formation. AB - Transcriptional profiling (TP) offers a powerful approach to identify genes activated during memory formation and, by inference, the molecular pathways involved. Trace eyeblink conditioning is well suited for the study of regional gene expression because it requires the hippocampus, whereas the highly parallel task, delay conditioning, does not. First, we determined when gene expression was most regulated during trace conditioning. Rats were exposed to 200 trials per day of paired and unpaired stimuli each day for 4 days. Changes in gene expression were most apparent 24 h after exposure to 200 trials. Therefore, we profiled gene expression in the hippocampus 24 h after 200 trials of trace eyeblink conditioning, on multiple arrays using additional animals. Of 1,186 genes on the filter array, seven genes met the statistical criteria and were also validated by real-time polymerase chain reaction. These genes were growth hormone (GH), c-kit receptor tyrosine kinase (c-kit), glutamate receptor, metabotropic 5 (mGluR5), nerve growth factor-beta (NGF-beta), Jun oncogene (c-Jun), transmembrane receptor Unc5H1 (UNC5H1), and transmembrane receptor Unc5H2 (UNC5H2). All these genes, except for GH, were downregulated in response to trace conditioning. GH was upregulated; therefore, we also validated the downregulation of the GH inhibitor, somatostatin (SST), even though it just failed to meet criteria on the arrays. By during situ hybridization, GH was expressed throughout the cell layers of the hippocampus in response to trace conditioning. None of the genes regulated in trace eyeblink conditioning were similarly affected by delay conditioning, a task that does not require the hippocampus. These findings demonstrate that transcriptional profiling can exhibit a repertoire of genes sensitive to the formation of hippocampal-dependent associative memories. PMID- 12542235 TI - A novel codification scheme based on the "VITAL" and "DICOM" Standards for telemedicine applications. AB - The field of interest discussed in this study is a novel codification scheme for (vital signs) medical device communication and patient monitoring data interchange, into the context of effective home care service provisioning. With medical technology having developed in isolation and major manufacturers developing their own proprietary communication protocols, which preclude connection to devices from different manufacturers, and with healthcare trends having evolved, pointing to primary care, telecare and home care monitoring, there is an increasing need for technical standardization in healthcare environments and the development of protocols that enable communication in a structured and open way. In this study, a novel codification scheme has been developed, based on two healthcare informatics standards, the VITAL and DICOM Sup. 30, in addressing the robust interchange of waveform and medical data for a home care application. Based on this scheme, we created a real-time facility, consisting of a base unit and a telemedicine (mobile) unit, that enables home telemonitoring, by installing the telemedicine unit at the patient's home while the base unit remains at the physician's office or hospital. The system allows the transmission of vital biosignals (3-lead ECG, pulse rate, blood pressure and SpO2) of the patient. This paper presents an object-oriented design with unified modeling language (UML) of a class hierarchy for exchanging the acquired medical data and performing alert management, and investigates the applicability of the proposed scheme into a commercial patient-connected medical device, thus addressing service and functionality requirements with focus on home-care applications. The system has been validated for technical performance over several telecommunication means and for clinical validity via real patient involved pilot trials. PMID- 12542236 TI - DIABCARD CCMIS--a portable and scalable CPR for diabetes care. AB - The aim of this paper is to introduce the main software module of the DIABCARD Chip Card Medical Information System (DIABCARD CCMIS) that provides an online, portable diabetes medical record information system based on a high performance object-oriented rapid application development language such as Borland Delphi. A chip card based medical information system was developed as a good possibility to create a portable electronic patient record. In particular the patient data card makes the up-to-date patient's record available whenever needed. The developed DIABCARD Core System, described in this paper, includes a patient record management system that has the ability to handle topics such as administrative and medical data, medical anamnesis, and physical examination data. Issues tackled were simplicity, data security and reporting, customization, and internationalization. Especially for the two last issues (customization and internationalization) a novel approach based on using native initialization table files is presented. Proper care has been addressed during the development of the software modules for matters of security, data integrity and confidentiality. PMID- 12542237 TI - Virtual center for renal support: technological approach to patient physiological image. AB - The patient physiological image (PPI) is a novel concept which manages the knowledge of the virtual center for renal support (VCRS), currently being developed by the Biomedical Engineering Group of the University of Seville. PPI is a virtual "replica" of the patient, built by means of a mathematical model, which represents several physiological subsystems of a renal patient. From a technical point of view, PPI is a component-oriented software module based on cutting-edge modeling and simulation technology. This paper provides a methodological and technological approach to the PPI. Computational architecture of PPI-based VCRS is also described. This is a multi-tier and multi-protocol system. Data are managed by several ORDBMS instances. Communications design is based on the virtual private network (VPN) concept. Renal patients have a minimum reliable access to the VCRS through a public switch telephone network--X.25 gateway. Design complies with the universal access requirement, allowing an efficient and inexpensive connection even in rural environments and reducing computational requirements in the patient's remote access unit. VCRS provides support for renal patients' healthcare, increasing the quality and quantity of monitored biomedical signals, predicting events as hypotension or low dialysis dose, assisting further to avoid them by an online therapy modification and easing diagnostic tasks. An online therapy adjustment experiment simulation is presented. Finally, the presented system serves as a computational aid for research in renal physiology. This is achieved by an open and reusable modeling and simulation architecture which allows the interaction among models and data from different scales and computer platforms, and a faster transference of investigation models toward clinical applications. PMID- 12542238 TI - Multimedia telehomecare system using standard TV set. AB - Nowadays, there are a very large number of patients that need specific health support at home. The deployment of broadband communication networks is making feasible the provision of home care services with a proper quality of service. This paper presents a telehomecare multimedia platform that runs over integrated services digital network and internet protocol using videoconferencing standards H.320 and H.323, and standard TV set for patient interaction. This platform allows online remote monitoring: ECG, heart sound, blood pressure. Usability, affordability, and interoperability were considered for the design and development of its hardware and software components. A first evaluation of technical and usability aspects were carried forward with 52 patients of a private clinic and 10 students in the University. Results show a high rate (mean = 4.33, standard deviation--SD = 1.63 in a five-points Likert scale) in the global perception of users on the quality of images, voice, and feeling of virtual presence. PMID- 12542239 TI - ATMTN: a telemammography network architecture. AB - One of the goals of the National Cancer Institute (NCI) to reach more than 80% of eligible women in mammography screening by the year 2000 yet remains as a challenge. In fact, a recent medical report reveals that while other types of cancer are experiencing negative growth, breast cancer has been the only one with a positive growth rate over the last few years. This is primarily due to the fact that 1) examination process is a complex and lengthy one and 2) it is not available to the majority of women who live in remote sites. Currently for mammography screening, women have to go to doctors or cancer centers/hospitals annually while high-risk patients may have to visit more often. One way to resolve these problems is by the use of advanced networking technologies and signal processing algorithms. On one hand, software modules can help detect, with high precision, true negatives (TN), while marking true positives (TP) for further investigation. Unavoidably, in this process some false negatives (FN) will be generated that are potentially life threatening; however, inclusion of the detection software improves the TP detection and, hence, reduces FNs drastically. Since TNs are the majority of examinations on a randomly selected population, this first step reduces the load on radiologists by a tremendous amount. On the other hand, high-speed networking equipment can accelerate the required clinic-lab connection and make detection, segmentation, and image enhancement algorithms readily available to the radiologists. This will bring the breast cancer care, caregiver, and the facilities to the patients and expand diagnostics and treatment to the remote sites. This research describes asynchronous transfer mode telemammography network (ATMTN) architecture for real time, online screening, detection and diagnosis of breast cancer. ATMTN is a unique high-speed network integrated with automatic robust computer-assisted diagnosis-detection/digital signal processing (CAD/DSP) methods for mass detection, region of interest (ROI) compression algorithms using Digital Imaging and Communications in Medicine (DICOM) 3.0 medical image standard. While ATMTN has the advantage of higher penetration for cancer screening, it provides the diagnosis with higher efficiency, better accuracy and potentially lower cost. This paper presents the development of the infrastructure and algorithm design for ATMTN-based telemammography. The research goals involved: 1) networking stations for telemammography to demonstrate, evaluate, and validate technologies and methods for delivering mammography screening services via high-speed (155 MB/s) links, performing real-time network-transmitted, high-resolution mammograms for immediate diagnosis as a "second opinion" strategy; 2) development of object oriented compression methods for storage, retrieval and transmission of mammograms; 3) inclusion and optimization of detection algorithms for identification of normal images in different resolutions to increase the speed and effectiveness of telemammography as a "second opinion" strategy; 4) resolving the compatibility issues between images from different equipment (DICOM standards); and 5) optimization of an integrated ATMTN with adaptive CAD/DSP methods that are robust for large image databases and input sources. PMID- 12542240 TI - Real-time collaborative environment for radiation treatment planning virtual simulation. AB - A virtual simulation (VS) system is a software system that enables the delineation of anatomical structures, and the placement of irradiation fields for the purpose of radiation treatment planning, making use of patient's tomographic data, instead of the real patient. Since patient's tomographic data can be communicated between distinct radiotherapy departments, collaborative work on VS, connecting remote health care professionals, becomes feasible. In this paper, an environment enabling real-time collaboration on VS is presented. The environment architecture is based on both offline and online communication of data under a secure framework and can be directly integrated into the infrastructure of a radiotherapy department. The online collaboration relies on the simultaneous execution of all actions at both collaborating sites, and prerequisites the offline communication of the data set on which the collaboration will be performed. Analytical description of the custom-made layered service, which supports the offline communication is given, along with a detailed presentation of the secure management of messages, which enables the real-time collaboration. The technical evaluation of the environment highlights the effectiveness of the proposed methodology, since real-time secure collaboration on VS is achieved. PMID- 12542241 TI - Asynchronous web-based patient-centered home telemedicine system. AB - A web-based system for asynchronous multimedia messaging between shoulder replacement surgery patients at home and their surgeons was developed and tested. A web browser plug-in simplifies the process of capturing video and transferring it to a web site for novice computer users. The design of the video capture plug in can be reused to acquire and securely transfer any type of data over the web. For example, readings from home biosensor instruments (e.g., glucometers and spirometers) that can be connected to a personal computer can be transferred to a home telemedicine web site. Both patients and doctors can access this web site to monitor health status longitudinally. Six patients, whose familiarity with computers ranged from no experience to expert users, used the system. All of the subjects were able to use the system to check treatment reminders and to send at least one message with video to their surgeons. The surgeons monitored the system regularly and always responded to messages within 24 h during the six-month trial period. PMID- 12542242 TI - A system for automatic measurement of circadian activity deviations in telemedicine. AB - A system for the automatic measurement of the circadian activity deviations in telemedicine has been developed within the framework of a "Health Integrated Smart Home Information System" (HIS2). HIS2 is an experimental platform for the evaluation and the development of technologies in order to ensure the security and quality of life for patients who need home based medical monitoring. Location sensors are placed in each room of the HIS2, allowing the monitoring of patient's successive activity phases within the patient's home environment. We proceeded with a sampling in an hourly schedule to detect weak rhythmic variations. Based on numerous measurements, we established a mean value with confidence limits. These also allowed us to define a zone within which the patient's activity is qualified to be "predictable." Alerts are set off if the patient's activity deviates from this zone. PMID- 12542243 TI - New approaches toward the fully digital integrated management of a burn unit. AB - In this paper, the design of an application that allows the integrated management of a burn unit is reported. Starting with the problems associated with the current procedures, technical solutions are found from the requirements demanded by the specialists. The major design considerations and implementation details are outlined. Special attention is devoted to the prescription of drugs and inventory control, as well as reducing the time that healthcare professionals spend in administrative tasks. The developed implementation is an example of a low-cost system suitable for adoption in a wide range of units in a hospital organization. PMID- 12542244 TI - Mobile access to biological databases on the Internet. AB - We have developed a new way of accessing biological databases and bioinformatics applications on the Internet. This new service, bioinformatics wireless application protocol (BioWAP) service, which is accessible by mobile devices makes it possible to access bioinformatics services, where normal PC or personal digital assistant (PDA) connections are not feasible. The BioWAP service includes major biological databases and applications demonstrating a simple method of implementing WAP interfaces to uncompliant applications, i.e. the applications that are not WAP or Internet based. The BioWAP service can be browsed with any WAP terminal. PMID- 12542245 TI - Recent advances in the molecular diagnosis of tuberculosis. AB - To date, the diagnosis of tuberculosis has not improved significantly and still relies heavily on staining and culture of sputum or other clinical specimens which were developed more than 100 years ago. Staining does not differentiate tuberculosis from other mycobacterial infections, and culture requires at least 4 to 8 weeks. These are the major problems faced by tuberculosis control programs. In response to this demand, new rapid diagnostic methods are urgently sought. In recent years, much hope has been laid on the development of molecular techniques in the routine tuberculosis laboratory. This review concentrates on 4 techniques that are increasingly used in clinical laboratories: polymerase chain reaction to detect mycobacterial DNA in clinical specimens, nucleic acid probes to identify culture, restriction fragment length polymorphism analysis to compare strains for epidemiologic purposes, and genetic-base susceptibility testing methods for rapid detection of drug resistance. Finally, the increase in the use of clinically useful molecular biological techniques that affect turnaround time, length of stay, and patient outcome, and reduce overall hospitalization costs will continue until universal standardization for molecular diagnostic procedures are provided. At present, conventional methods should not be replaced by novel methods until the latter are shown to be of equal or greater sensitivity, specificity, reliability, and user-friendliness. However, it is expected that the newly developed molecular techniques will complement our armamentarium of diagnostic tools in the detection of tuberculosis. It is also expected that clinical protocols based on molecular methods will increase the chances for cure by selecting the most appropriate treatment and improving the quality of life of tuberculosis patients. PMID- 12542246 TI - Prognosis of tuberculous meningitis in adults in the era of modern antituberculous chemotherapy. AB - This study reviewed the clinical manifestations and outcome of tuberculous meningitis in the era of modern antituberculous chemotherapy and applied these data in assessing the role of clinical staging evaluated 30 days after treatment in predicting long-term outcome. A total of 41 adult patients with tuberculous meningitis hospitalized at a university hospital in Taiwan from June 1994 through August 1999 were included in this retrospective study. Their age ranged from 16 to 80 years (median, 41 years), and 17 (41.5%) patients had had a variety of underlying immunocompromising diseases. Fever (90%), headache (75.6%), neck stiffness (68.3%), altered consciousness (26.8%), and nausea and/or vomiting (26.8%) were the leading initial presentations. During the treatment course, 19 patients experienced new neurologic complications. The overall case fatality rate was 9.8% and morbidity rate 56.1%. More advanced clinical stage evaluated at 30 days after initiation of antituberculous chemotherapy and positive cerebrospinal fluid culture for Mycobacterium tuberculosis were the only 2 factors significantly associated with a worse long-term prognosis. Results indicate that tuberculous meningitis is associated with a high morbidity, consisting of minor and major neurologic sequelae, despite modern antituberculous chemotherapy. In addition, more advanced clinical staging evaluated at 30 days after the start of antituberculous chemotherapy and a positive cerebrospinal fluid culture for M. tuberculosis were associated with a poor prognosis. PMID- 12542247 TI - Phylogenetic analysis of Newcastle disease virus in Taiwan. AB - The virulent forms of Newcastle disease virus cause a devastating disease of poultry. Between 1998 and 2000, sporadic outbreaks of Newcastle disease occurred in Taiwan despite vaccination. The causes of the failure of the vaccination remain unclear. The purpose of this study was to investigate the possible factors causing these outbreaks by serologic and virologic methods. Anti-Newcastle disease virus hemagglutination-inhibition titers were measured for serum samples obtained from a breeder farm and a broiler farm. The serologic data showed continued presence of virulent Newcastle disease viruses in the field during inter-outbreak periods. Phylogenetic analysis demonstrated that the field virulent Newcastle disease viruses were genetically similar and were grouped into genotype VIIa. Efficacy testing by virulent Newcastle disease virus challenge revealed that the vaccines used were effective for protecting chickens from infections. This investigation demonstrated that the Newcastle disease virus strain can spread quickly and widely throughout a large geographic area, and that the sporadic cases originate from virulent Newcastle disease viruses present in the field. PMID- 12542249 TI - Monitoring the hygene of chicken hatcheries in Taiwan during 1999-2001. AB - Microorganism contamination in hatcheries and eggs has a serious impact on the viability and quality of chicks as well as on the overall growth performance of chickens. Microbiological agents are present in the fluff when chicks hatch. Detecting microorganisms in fluff is a convenient method for evaluating the hygienic status in a hatchery. Fluff samples from 31 hatcheries collected over 3 years were tested for the total bacterial count, the presence of Salmonella spp., and fungus to evaluate the hygienic status of hatcheries in Taiwan from 1999 through 2001. The total bacterial score from the fluff samples was calculated and expressed as a bacterial score in a log scale. Most hatcheries had a bacterial count ranged from scale 1 to 3. Among the hatcheries, 13% to 29% were contaminated with Salmonella spp.; and 33% to 73% were contaminated with fungi in different quarters. The third quarter of each year was the most contaminated period (p<0.01). According to the data obtained from the fluff tests, hatcheries keep their hygienic status and supply good quality chicks by cleaning and disinfecting. PMID- 12542248 TI - Characterization of a highly attenuated Salmonella enterica serovar Typhimurium mutant strain. AB - Salmonella enterica serovar Typhimurium ATCC 13311 is virulent at a dose as low as 10(2) colony-forming units when administered intraperitoneally to BALB/c mice. In order to develop highly attenuated mutant strain through the combination of 2 phenotypically attenuated markers, we constructed a number of amino acid requiring auxotrophic strains of S. enterica serovar Typhimurium by means of UV induced mutations. One of them, strain NDMC-B1, was highly attenuated for mice, with an LD50-value of 6 and 3 log units lower for mice than the wild-type strain and S. enterica serovar Typhimurium aroA strain, respectively. This strain still contained the Salmonella O- and H-antigens but had a requirement for cysteine and was unable to utilize citrate as its sole carbon source. NDMC-B1 colonized the gut-associated lymphoid tissue more efficiently than the wild-type strain, but its capacities to colonize spleen and liver were significantly reduced. Mice intraperitoneally or orally vaccinated with NDMC-B1 were highly protected against either an intraperitoneal challenge with 10(6) colony-forming units or an oral challenge with 10(9) colony-forming units of the wild-type strain. Taken together, the results illustrate that through the combination of 2 independently phenotypical attenuating markers, the requirement for cysteine and the inability to use citrate, we have successfully constructed a highly attenuated, stable, and immunogenic S. enterica serovar Typhimurium vaccine strain which can induce protective immunity in a mouse model against lethal challenge of wild-type strain. PMID- 12542250 TI - Phenotypes and genotypes of vancomycin-resistant enterococci isolated during long term follow-up in a patient with recurrent bacteremia and colonization. AB - Twenty-seven isolates of vancomycin-resistant enterococci were obtained at monthly intervals from a bed-ridden man with hypoxic encephalopathy. During the 28-month period of the patient's hospitalization, 3 episodes of bacteremia and one episode of catheter-related infection caused by vancomycin-resistant enterococci occurred. Rectal swabs showed colonization of vancomycin-resistant enterococci for more than 2 years. Three months after termination of antimicrobial therapy, the rectal colonization for vancomycin-resistant enterococci was eradicated. Four species (Enterococcus faecium, Enterococcus gallinarum, Enterococcus faecalis, and Enterococcus casseliflavus) were identified among the 27 vancomycin-resistant enterococcus isolates. Three non clonal related patterns were found among 17 strains of E. faecium by pulsed-field gel electrophoresis. All of the 3 E. faecalis isolates were of the VanB phenotype, but of the vanA genotype. Linezolid had the most potent in vitro activity against these vancomycin-resistant enterococcus isolates, with minimum inhibitory concentrations >2 microg/mL. Eighty-five percent of these vancomycin resistant enterococcus isolates were susceptible to tetracycline and 66% were susceptible to quinupristin-dalfopristin. Although a high genetic correlation of E. faecium was identified in the patient with prolonged hospitalization, the isolation of 3 genetically unrelated colonized isolates suggested a lack of correlation between infection and colonization. Precautions against resistant organisms, adapted antibiotic policies, and elimination of patient carriage are useful for controlling the spread of vancomycin-resistant enterococci. PMID- 12542251 TI - Clinical significance of Candida species isolated from cerebrospinal fluid. AB - Candida species have only rarely been isolated from the cerebrospinal fluid. Some of these isolates are true pathogens with a high morbidity and mortality, while others are only colonizers or contaminants. Spontaneous recovery from Candida meningitis has also been reported. The purpose of this study was to identify factors indicative of patients positive for Candida from cerebrospinal fluid who should receive antifungal therapy. A total of 36 patients with a positive cerebrospinal fluid culture for Candida were included in this retrospective analysis. Seventeen of these patients had received antifungal therapy under the impression of Candida meningitis. The differences in the case fatality rate and the duration of hospitalization between the antifungal treatment and untreated groups were statistically significant (p<0.04 and p<0.005, respectively). Statistical analysis showed a significant difference in the percentage of patients with previous ventricular shunt (14/17 vs 8/19, p<0.05), central venous line in situ (9/17 vs 1/19, p<0.005), multiple positive cerebrospinal fluid culture (11/17 vs 1/19, p<0.0005), isolation of Candida at least 20 days after hospitalization in adults (6/7 vs 1/13, p<0.005), and cerebrospinal fluid white blood cell count (median 77 vs 2 /mm3, p<0.005). These results suggest that antifungal agent should be initiated promptly in patients whose cerebrospinal fluid is positive for Candida and who have one or more of the identified risk factors. PMID- 12542252 TI - Bacteroides fragilis bacteremia associated with portal vein and superior mesentery vein thrombosis secondary to antithrombin III and protein C deficiency: a case report. AB - Hypercoagulability is one of the causes of portal vein and superior mesentery vein thrombosis. We report a case of Bacteroides fragilis bacteremia associated with portal vein and superior mesentery vein thrombosis secondary to antithrombin III and protein C deficiency. The patient presented with high fever for more than 3 weeks. Abdominal sonography revealed a liver cyst of 1.7 cm in diameter over segment 4 and a renal stone of 0.7 cm in size over the lower portion of the right kidney but no evidence of hydronephrosis. Elevation of liver enzymes was also noted. Intermittent fever was noted despite treatment with ceftriaxone and doxycycline. On Day 15 of hospitalization, blood culture revealed B. fragilis, which prompted further investigation of the source of intraabdominal and pelvic infection. Abdominal computed tomography revealed portal vein and superior mesentery vein thrombosis. Endoscopic studies of the gastrointestinal tract showed no tumor or diverticulum. Study of coagulation factors disclosed deficiency of antithrombin III and protein C. Clinicians should remain aware of the need to promptly search for a portal or mesentery vein thrombosis in cases of Bacteroides bacteremia of unknown origin. PMID- 12542253 TI - Steroid refractory interstitial pneumonitis in a patient with juvenile dermatomyositis. AB - Interstitial pneumonitis is a severe complication of juvenile dermatomyositis. We report a 4-year-old girl with juvenile dermatomyositis. Coughing, shortness of breath, and general malaise developed during steroid treatment. The histology of her lung biopsy is compatible with interstitial pneumonitis. Aggressive treatment including intravenous methylprednisolone pulse therapy, intravenous immunoglobulin, and oral cyclosporin all failed. Creatinine phosphokinase level was within the normal range during the disease course. The clinical features are discussed and the importance of a differential diagnosis of interstitial pneumonitis in patients with juvenile dermatomyositis is emphasized. PMID- 12542254 TI - Neonatal lupus erythematosus with recurrent pancytopenia: a case report. AB - Neonatal lupus erythematosus is an immune-mediated disease associated with the transplacental passage of maternal autoantibodies, primarily anti-Ro (SS-A) and anti-La (SS-B). The major clinical manifestations are isolated congenital heart block and cutaneous lupus lesions. Other symptoms include hematologic and hepatic abnormalities. We report a male neonate, born to a mother with Sjogren's syndrome, who experienced cutaneous lupus lesions and 2 episodes of pancytopenia. Both anti-Ro and anti-La antibodies were positive in infant and mother. PMID- 12542255 TI - Interleukin-10 levels in Epstein-Barr virus-associated nasopharyngeal carcinoma. AB - The aim of this study was to determine the levels of interleukin-10 in patients with nasopharyngeal carcinoma. Both biopsies and sera were obtained from patients with nasopharyngeal carcinoma as well as Epstein-Barr virus-seronegative patients as a control. Nasopharyngeal carcinoma patients were classified using the World Health Organization pathological assessment and clinical staging of nasopharyngeal carcinoma. The numbers of interleukin-10 positive cells and the levels of serum interleukin-10 were assessed by immunohistochemical methods and enzyme-linked immunosorbent assay, respectively. The levels of serum interleukin 10 were determined by enzyme-linked immunosorbent assay. The results showed that the number of interleukin-10 positive cells and serum interleukin-10 levels were significantly increased in patients with nasopharyngeal carcinoma-World Health Organization type III and with clinical late stage (p<0.05), suggesting that interleukin-10 may have a crucial role in the progression of nasopharyngeal carcinoma. PMID- 12542256 TI - Analysis of the baseplate region of phage AR1 that specifically infects Escherichia coli O157:H7. AB - Phage AR1 that specifically infects Escherichia coli O157:H7 has been characterized previously for its primary host contact molecule, the long tail fiber. To strengthen its attachment to the host bacteria, the phage may use its baseplate for a second contact. This study investigated the characteristics of AR1 baseplate by deducing the responsible genes including wac and genes 9 to 15. The wedge pin g11 and the short tail fiber g12 varied the most, whereas the others were highly conserved compared with their counterparts in T4. The degrees of gene conservation correlated well with the known functions of individual gene products. PMID- 12542257 TI - Guidelines for antimicrobial therapy of acute upper respiratory tract infections in Taiwan. PMID- 12542259 TI - The interaction of rhinal cortex and hippocampus in human declarative memory formation. AB - Human declarative memory formation crucially depends on processes within the medial temporal lobe (MTL). These processes can be monitored in real-time by recordings from depth electrodes implanted in the MTL of patients with epilepsy who undergo presurgical evaluation. In our studies, patients performed a word memorization task during depth EEG recording. Afterwards, the difference between event-related potentials (ERPs) corresponding to subsequently remembered versus forgotten words was analyzed. These kind of studies revealed that successful memory encoding is characterized by an early process generated by the rhinal cortex within 300 ms following stimulus onset. This rhinal process precedes a hippocampal process, which starts about 200 ms later. Further investigation revealed that the rhinal process seems to be a correlate of semantic preprocessing which supports memory formation, whereas the hippocampal process appears to be a correlate of an exclusively mnemonic operation. These studies yielded only indirect evidence for an interaction of rhinal cortex and hippocampus. Direct evidence for a memory related cooperation between both structures, however, has been found in a study analyzing so called gamma activity, EEG oscillations of around 40 Hz. This investigation showed that successful as opposed to unsuccessful memory formation is accompanied by an initial enhancement of rhinal-hippocampal phase synchronization, which is followed by a later desynchronization. Present knowledge about the function of phase synchronized gamma activity suggests that this phase coupling and decoupling initiates and later terminates communication between the two MTL structures. Phase synchronized rhinal-hippocampal gamma activity may, moreover, accomplish Hebbian synaptic modifications and thus provide an initial step of declarative memory formation on the synaptic level. PMID- 12542258 TI - Cognitive processing in the primary visual cortex: from perception to memory. AB - The primary visual cortex is the first cortical area of the visual system that receives information from the external visual world. Based on the receptive field characteristics of the neurons in this area, it has been assumed that the primary visual cortex is a pure sensory area extracting basic elements of the visual scene. This information is then subsequently further processed upstream in the higher-order visual areas and provides us with perception and storage of the visual environment. However, recent findings show that such neural implementations are observed in the primary visual cortex. These neural correlates are expressed by the modulated activity of the late response of a neuron to a stimulus, and most likely depend on recurrent interactions between several areas of the visual system. This favors the concept of a distributed nature of visual processing in perceptual organization. PMID- 12542261 TI - Influence of enriched environment on spatial learning following cerebral insult. AB - The fact that our brain is continuously shaped by the environment and experience presents many challenges. It was noted several decades ago that exposure of a laboratory rat to a complex environment alters its brain and improves problem solving in complex tasks. Rats that are housed in an enriched environment can better compensate for brain lesion-induced deficits and resist neurodegeneration. In this article, the role of an enriched environment on brain plasticity following experimentally-induced lesions is discussed. PMID- 12542260 TI - Information processes in the primate prefrontal cortex in relation to working memory processes. AB - Working memory is a mechanism for short-term active storage of information as well as for processing stored information. Although evidence for neuronal mechanisms of temporary storage of information has accumulated for the prefrontal cortex, little is known about neuronal mechanisms for processing information. To understand how information is processed by prefrontal neurons, we first need to know what information is represented by single-neuron activity, and then examine how information represented by single-neuron activity or a population of activities changes along the temporal sequence of the trial. By examining task related single-neuron activities while monkeys performed various working memory tasks, delay-period activity observed in the prefrontal cortex is considered to be a neuronal correlate of the mechanism for temporary active storage of information. Delay-period activity represents a variety of information including the spatial position and the physical feature of the stimulus, the forthcoming behavioral response, the quality of reward that the subject would receive, the difference of the task, or the rule of the task. Although delay-period activity could represent this variety of information, the information represented by delay period activity is only the information relevant for task performance. In addition, using complex conditional tasks, delay-period activity has been shown to represent several kinds of information simultaneously. Based on these results, we examined how information represented by a population of prefrontal activities changes along the temporal sequence of the trial. Using two kinds of oculomotor delayed-response tasks, we first identified what information each task-related activity represents. Then, using population vector analysis, we could not only visualize information represented by a population of prefrontal activities, but also demonstrate the temporal change of information represented by a population of prefrontal activities. These attempts are important to understand information processes for working memory. PMID- 12542262 TI - Sedentary life impairs self-reparative processes in the brain: the role of serum insulin-like growth factor-I. AB - Regular exercise has long being recognized as an important contributor to appropriate health status and is currently recommended to reduce the incidence of many diseases. More recent is the notion that sedentary life may also be a risk factor for neurodegenerative diseases even though for the last decade the beneficial effects of exercise on brain function have been widely documented. In the brain, exercise exerts both acute and long-term changes that can be interpreted as beneficial, such as increased levels of various neurotrophic factors or enhanced cognition. However, the signals involved in exercise-induced changes in the brain are not yet well known. It is generally thought that they arise from the periphery as a direct consequence of increased metabolic activity and aim to elicit adaptive changes in brain function. However, body-to-brain signaling induced by exercise also underlies a different aspect. Exercise induces changes in the brain that are essential for proper brain function. In this view, sedentarism, a relatively new cultural trait, negates the beneficial effects of exercise and paves the way to pathological derangement. A critical step in this process is exercise-induced uptake by the brain of insulin-like growth factor-I (IGF-I), a circulating hormone with potent neurotrophic activity. We summarize the evidence supporting the hypothesis that serum IGF-I is a neuroprotective hormone within a neuroprotective network modulated by physical activity. PMID- 12542263 TI - Is it only neurogenesis? AB - For a long time, the occurrence of neurogenesis in the adult mammalian brain was deemed non-existent or, at best, restricted to phylogenetically old brain regions. The pendulum of current opinion has now swung in the opposite direction with growing awareness that incorporation of labeled precursors into neuronal DNA occurs widely in the brain, and undergoes significant modulation with learning, different kinds of experiential inputs, and a number of physiological manipulations. A thorough review of the literature indicates that unscheduled DNA synthesis may significantly contribute to available evidence. Notably, data interpreted in terms of nerve cell turnover are more likely to reflect turnover of neuronal DNA, as suggested by earlier investigations. PMID- 12542264 TI - Veterinary surgeon to be struck off for false certification and unprofessional behaviour. PMID- 12542265 TI - Direction of the BVA: council agrees on planning the way forward. PMID- 12542266 TI - Comparative analyses of Pasteurella multocida strains associated with the ovine respiratory and vaginal tracts. AB - Thirty-five isolates of Pasteurella multocida from the vagina and respiratory tract of sheep were compared by analysing their capsular polysaccharide types and outer membrane protein profiles. The phylogenetic relationships of selected isolates with respect to reference strains of P. multocida were also determined by comparative 16S rRNA sequence analysis. Three capsular types, A, D and F, and three major outer membrane protein types were identified, and there were four different combinations of these characteristics which probably marked four individual clones of P. multocida. Strains representing three of these clones were recovered from cases of ovine pneumonia, whereas isolates of the fourth clone were associated exclusively with the vagina of healthy ewes and the liver of a dead septicaemic lamb on the same farm. Analysis of the 16S rRNA sequences showed that there was 100 per cent identity between representative pneumonic isolates and reference strains of P. multocida subspecies galliseptica and P. multocida subspecies multocida. The 16S rRNA genes of representative vaginal and liver isolates from the same farm were identical but differed from the other strains at one nucleotide position, providing strong evidence that the vaginal and liver isolates represent a distinct subpopulation of P. multocida. PMID- 12542267 TI - Reduced dosage of ketoprofen for the short-term and long-term treatment of joint pain in dogs. AB - Two studies were conducted under laboratory conditions with 16 dogs to investigate the analgesic effectiveness of a low dose of ketoprofen in a short term sodium urate crystal-induced synovitis model of arthritis. The effect of the treatment, defined as the improvement in peak vertical force weight bearing was evaluated in the first study at three dose levels. A single oral dose of 0.25 mg/kg ketoprofen was significantly better (P < 0.01) than the control (0 mg), but doses of 0.5 and 0.75 mg/kg did not improve the dogs' weight bearing further. The second study investigated the efficacy and safety of the 0.25 mg/kg dose administered daily for 30 days. The beneficial effects of ketoprofen at this dose were constant, with the treated dogs bearing 89.1 per cent of the baseline vertical force four hours after the induction of arthritis on day 1 and 92.2 per cent on day 29, compared with 42 per cent and 34 per cent of the baseline in the untreated dogs. No gastrointestinal or other side effects were observed during the treatment. PMID- 12542268 TI - Patterns of exposure to Lawsonia intracellularis infection on European pig farms. AB - A serological investigation was made of the patterns of exposure of pigs to Lawsonia intracellularis, the causative agent of proliferative enteropathy (ileitis), on farms in France and Spain. Blood samples from groups of adult female pigs in breeding programmes and from postweaning pigs were monitored, the latter every month for five months, by a L. intracellularis-specific immunofluorescence seroassay. Four of 33 farms monitored in France (12 per cent) and three of 29 farms monitored in Spain (10.3 per cent) remained free of clinical signs and seronegative throughout the study. The postweaning pigs on all of the remaining French farms and on 20 of the 26 remaining Spanish farms had a pattern of infection characterised by seroconversion in the grower period, generally between eight and 16 weeks of age. The seroprevalence in these groups ranged from 8 to 20 per cent. On all of these farms at least 15 per cent of the breeding females tested were seropositive, and the farms were under similar management systems, with a continuous flow of pigs or between buildings on one site, so-called 'one site, farrow-to-finish'. On the six remaining Spanish farms, under two management groups, a multiple-site system was used, with the piglets being separated from the adults at weaning and moved to a separate location. On three of these farms, the pattern of infection was characterised by seroconversion later in the finisher period, at between 16 and 20 weeks of age, and none of the breeding females was seropositive. On the three other multiple site farms the pattern of infection resembled that on the one-site farms. On all of the farms, the seroconversion of groups of pigs was frequently associated with clinical or subclinical signs of ileitis. PMID- 12542269 TI - Isoflurane to prolong medetomidine/ ketamine anaesthesia in six adult female chimpanzees (Pan troglodytes). AB - Six adult female chimpanzees (Pan troglodytes) were anaesthetised for the placement of intrauterine contraceptive devices, microchips for identification, routine blood sampling, and physical measurements. Anaesthesia was induced with medetomidine in combination with ketamine administered by intramuscular injection with a projectile syringe. Induction was smooth and rapid, but five of the animals were insufficiently relaxed for orotracheal intubation. The plane of anaesthesia was deepened by administering isoflurane delivered in oxygen and nitrous oxide, and general anaesthesia was maintained for up to 74 minutes. The action of medetomidine was reversed at the end of each procedure with atipamezole, and the animals recovered smoothly and uneventfully. PMID- 12542270 TI - Antibacterial activity of ceftiofur-impregnated polymethylmethacrylate beads following manufacture, storage and sterilisation. PMID- 12542271 TI - Helminths found in hedgehogs (Erinaceus europaeus) in three areas of Italy. PMID- 12542272 TI - Mastitis and somatic cell counts. PMID- 12542274 TI - Haemorrhagic enteritis in turkeys. PMID- 12542275 TI - International sequestration experiment and conventions for the protection of the marine environment. PMID- 12542276 TI - Is a weakening of U.S. environmental policy ahead? PMID- 12542277 TI - Can private-sector funds resolve U.S. water woes? PMID- 12542278 TI - Does hydraulic fracturing harm groundwater? PMID- 12542279 TI - Hanford's vitrification challenge. PMID- 12542280 TI - Characterizing aquatic dissolved organic matter. PMID- 12542281 TI - Developing clean coal technology. PMID- 12542282 TI - Field kits fail to provide accurate measure of arsenic in groundwater. PMID- 12542283 TI - Levels of extractable organohalogens in pine needles in China. AB - Pine needles can accumulate organohalogens from the ambient air, which are, hence, able to serve as a biomonitor to evaluate the levels of organochlorinated contaminants in the atmospheric environment. Extractable organochlorine (EOCl), the most abundant of the extractable organohalogens (EOX) in environmental samples, has received much attention as a parameter for evaluation of total contamination levels of organochlorinated compounds (OCs). However, few data concerning EOCl in vegetation are available. In this study, we selected pine needles from 17 different areas in China as a sampling matrix to reflectthe regional distribution of OCs. EOX (EOX = EOCl + EOBr + EOI) were measured by instrument neutron activation analysis for their concentrations and distribution in pine needles. The concentrations of EOX were on the order of EOCl >> EOBr > EOI. About 5-38% of EOCl remained as sulfuric acid-resistant organochlorine (EPOCl). The relatively high concentrations of EPOCl in pine needles from remote areas suggested that EPOCl mainly originated from long-range atmospheric transport and contaminated soil. The relative proportions of the known organochlorines (such as HCHs, DDTs, aldrin, heptachlor, and chlordanes) to total EOCl and EPOCl were 0.3-5.2% and 1.4-19.8%, respectively, which implied that a major portion of the EPOCl measured in pine needles was unknown. The EPOX accumulation rates were preliminarily estimated under the natural condition, which suggested that the "young" needle accumulated EPOX more quickly than the "old" and more than 94% of EPOX was accumulated at the first year of pine needles. PMID- 12542284 TI - A predictive tool for emissions from heavy-duty diesel vehicles. AB - Traditional emissions inventories for trucks and buses have relied on diesel engine emissions certification data, in units of g/bhp-hr, processed to yield a value in g/mile without a detailed accounting of the vehicle activity. Research has revealed a variety of other options for inventory prediction, including the use of emissions factors based upon instantaneous engine power and instantaneous vehicle behavior. The objective of this paper is to provide tabular factors for use with vehicle activity information to describe the instantaneous emissions from each heavy-duty vehicle considered. To produce these tables, a large body of data was obtained from the research efforts of the West Virginia University Transportable Heavy Duty Emissions Testing Laboratories (TransLabs). These data were available as continuous records of vehicle speed (hence also acceleration), vehicle power, and emissions of carbon monoxide (CO), oxides of nitrogen (NOx), and hydrocarbons (HC). Data for particulate matter (PM) were available only as a composite value for a whole vehicle test cycle, but using a best effort approach, the PM was distributed in time in proportion to the CO. Emissions values, in g/sec, were binned according to the speed and acceleration of a vehicle, and it was shown that the emissions could be predicted with reasonable accuracy by applying this table to the original speed and acceleration data. The test cycle used was found to have a significant effect on the emissions value predicted. Tables were created for vehicles grouped by type (large transit buses, small transit buses, and tractor-trailers) and by range of model year. These model year ranges were bounded by U.S. national changes in emissions standards. The result is that a suite of tables is available for application to emissions predictions for trucks and buses with known activity, or as modeled by TRANSIMS, a vehicle activity simulation model from Los Alamos National Laboratories. PMID- 12542285 TI - Alkyl amides and nitriles as novel tracers for biomass burning. AB - The occurrence of n-alkanoic acids, amides, and nitriles in samples of aerosol particulate matter from Kuala Lumpur and Santiago suggests that emissions from cooking and biomass burning are the primary sources of these organic markers in the atmosphere. It is proposed that fatty acids react with ammonia during biomass burning or combustion to produce amides and nitriles, which can be applied as useful biomarker tracers. To test this hypothesis, nonadecanoic acid and hexadecanamide were used as reactants in hydrous pyrolysis experiments. These experiments produced amides and nitriles and indicated that ammonia is an essential agent in their formation. Thus amides and nitriles are of utility as indicators for input from combustion and biomass burning in the ambient atmosphere. PMID- 12542286 TI - Measurements of atmospheric mercury species at a coastal site in the Antarctic and over the south Atlantic Ocean during polar summer. AB - Mercury and many of its compounds behave exceptionally in the environment because of their volatility, capability for methylation, and subsequent biomagnification in contrast with most of the other heavy metals. Long-range atmospheric transport of elemental mercury, its transformation to more toxic methylmercury compounds, the ability of some to undergo photochemical reactions, and their bioaccumulation in the aquatic food chain have made it a subject of global research activities, even in polar regions. The first continuous high-time-resolution measurements of total gaseous mercury in the Antarctic covering a 12-month period were carried out at the German Antarctic research station Neumayer (70 degrees 39' S, 8 degrees 15' W) between January 2000 and February 2001. We recently reported that mercury depletion events (MDEs) occur in the Antarctic after polar sunrise, as was previously shown for Arctic sites. These events (MDEs) end suddenly during Antarctic summer. A possible explanation of this phenomenon is presented in this paper, showing that air masses originating from the sea-ice surface were a necessary prerequisite for the observations of depletion of atmospheric mercury at polar spring. Our extensive measurements at Neumayer of atmospheric mercury species during December 2000-February 2001 show that fast oxidation of gaseous elemental mercury leads to variable Hg0 concentrations during Antarctic summer, accompanied by elevated concentrations, up to more than 300 pg/m3, of reactive gaseous mercury. For the first time in the Southern Hemisphere, atmospheric mercury species measurements were also performed onboard of a research vessel, indicating the existence of homogeneous background concentrations over the south Atlantic Ocean. These new findings contain evidence for an enhanced oxidizing potential of the Antarctic atmosphere over the continent that needs to be considered for the interpretation of dynamic transformations of mercury during summertime. PMID- 12542287 TI - Effect of peat decomposition and mass loss on historic mercury records in peat bogs from patagonia. AB - Ombrotrophic peat bogs have been widely used to evaluate long-term records of atmospheric mercury (Hg) deposition. One of the major aims of these investigations is the estimation of the increase in atmospheric Hg fluxes during the industrial age compared to preindustrial fluxes. Comparability of Hg accumulation rates calculated from density, peat accumulation rates, and Hg concentrations requires linearity between these parameters. Peat formation is a dynamic process accompanied by intense mass loss and alteration of the organic material. Our investigations on three peat cores from the Magellanic Moorlands, Chile, indicate that Hg concentrations in peat strongly depend on peat humification. Moreover, differences in mass accumulation rates during peat evolution are not compensated by linear changes in density, peat accumulation, or Hg concentrations. We suggest that Hg accumulation rates be normalized to carbon accumulation rates to achieve comparability of Hg accumulation rates derived from differently altered peat sections. Normalization to the carbon accumulation rates reduces Hg accumulation rates in less degraded peat sections in the upper peat layers by factors of more than 2. Our results suggest that the increase in Hg deposition rates during modern times derived from ombrotrophic peat bogs are potentially overestimated if Hg accumulation rates are not corrected for mass accumulation rates. PMID- 12542288 TI - Estimating the natural background atmospheric deposition rate of mercury utilizing ombrotrophic bogs in southern Sweden. AB - A critical gap in the understanding of the global cycling of mercury is the limited data describing the natural background atmospheric deposition rate of mercury before the advent of pollution. Existing estimates of the natural deposition rate are typically about 2-5 microg of Hg m(-2) year(-1) (see, for example, Swain et al. Science 1992, 257, 784-787), based on studies that generally rely on short, 210Pb-dated lake sediment and peat cores that span the past 150 years. Analyses of mercury in long peat cores in southcentral Sweden indicate that natural mercury deposition rates in the period 4000-500 BP were lower, about 0.5-1 microg of Hg m(-2) year(-1). This suggests that recent mercury accumulation rates in the peat (15-25 microg of Hg m(-2) year(-1)) and measured atmospheric deposition rates of mercury in Sweden over the past 3 decades (5-30 microg of Hg m(-2) year(-1)) (Munthe et al. Water, Air, Soil Pollut.: Focus 2001, 1, 299-310) are at least an order of magnitude greater than the prepollution deposition rate, rather than representing only a 3-5-fold increase, as has generally been estimated. PMID- 12542289 TI - Polycyclic aromatic hydrocarbons in a semiaquatic plant and semipermeable membrane devices exposed to air in Thailand. AB - Semipermeable membrane devices (SPMDs) were deployed at six sites in the Bangkok region, Thailand, to investigate spatial variations in atmospheric concentrations of polycyclic aromatic hydrocarbons (PAHs). Sampling sites affected by various levels of traffic intensity were studied. In addition, PAH levels were determined in a common human food plant (water spinach) harvested from canals and ponds in the sampling areas. Significant differences in atmospheric PAH concentrations between sites were found, with 10 times higher PAH levels in the urban areas compared to the rural areas. Increasing concentrations of 1-methylphenanthrene relative to phenanthrene were found in the urban air close to the city center, indicating that traffic probably contributed to the higher PAH concentrations detected. Due to SPMD's passive sampling technique, their long-term operation and high ability to detect spatial differences, they proved to be suitable for semiquantitative field studies of PAHs. The PAH compounds sampled with SPMDs were mainly associated with gaseous PAHs, while both gas phase and particle-bound PAHs were detected in the plant samples. The relative abundance ratios of some PAHs in the plants were not well correlated with the ratios detected in the SPMDs, indicating that gas-phase exposure made low contribution to the PAH concentrations in the plants. However, similarities in the profiles of 3-ring PAHs between the SPMD and plant samples indicate that gas-phase exchange occurs between the atmosphere and the plants. PMID- 12542290 TI - High-resolution record of pyrogenic polycyclic aromatic hydrocarbon deposition during the 20th century. AB - A high-resolution record of polycyclic aromatic hydrocarbon (PAH) deposition in Rhode Island over the past approximately 180 years was constructed using a sediment core from the anoxic Pettaquamscutt River basin. The record showed significantly more structure than has hitherto been reported and revealed four distinct maxima in PAH flux. The characteristic increase in PAH flux at the turn of the 20th century was captured in detail, leading to an initial maximum prior to the Great Depression. The overall peak in PAH flux in the 1950s was followed by a maximum that immediately preceded the 1973 Organization of Petroleum Exporting Countries (OPEC) oil embargo. During the most recent portion of the record, an abrupt increase in PAH flux between 1996 and 1999 has been found to follow a period of near constant fluxes. Because source-diagnostic ratios indicate that petrogenic inputs are minor throughout the record, these trends are interpreted in terms of past variations in the magnitude and type of combustion processes. For the most recent PAH maximum, energy consumption data suggest that diesel fuel combustion, and hence traffic of heavier vehicles, is the most probable cause for the increase in PAH flux. Systematic variations in the relative abundance of individual PAHs in conjunction with the above changes in flux are interpreted in relation to the evolution of combustion processes. Coronene, retene, and perylene are notable exceptions, exhibiting unique down core profiles. PMID- 12542291 TI - Characterizing the emissions of polychlorinated dibenzo-p-dioxins and dibenzofurans from crematories and their impacts to the surrounding environment. AB - This study was set out to characterize PCDD/F emissions from crematories and assess their impacts on the surrounding environment. Two crematories located in southern Taiwan were investigated, including the one (C1) with no air pollution control device installed and the other (C2) installed with a bag filter. Results show the mean PCDD/F emissions (11% oxygen) from the stacks of Cl and C2 were 2.36 and 0.322 ng I-TEQ Nm(-3), respectively. The mean emission factors for C1 and C2 were 13.6 and 6.11 microg I-TEQ body(-1), respectively. The removal efficiency of the bag filter on PCDD/Fs was 55.1%. The estimated PCDD/F emission rate for all crematories in Taiwan was 0.838 g I-TEG yr(-1) accounting for 227% and 112% of the annual emissions from all medical waste incinerators and municipal waste incinerators, respectively. The above results indicate that PCDD/F emissions from crematories were quite significant. To assess the impact of PCDD/F emissions from a crematory to the surrounding environment, ambient air samples were collected from the downwind site of C1 with the maximum ground concentration. We found the estimated maximum ground concentration at the downwind site of C1 (= 0.521 pg I-TEQ Nm(-3)) was much higherthan thatfound atthe background, rural area, residential area, urban area, and industrial area (= 0.006, 0.023, 0.052, 0.093, and 0.190 pg I-TEG Nm(-3), respectively). The above results suggestthat PCDD/F emissions from a crematory did significantly affect its surrounding environment. In conclusion, a proper control strategy should be taken immediately in order to eliminate PCDD/F emissions from crematories. PMID- 12542292 TI - Kinetic speciation of Co(II), Ni(II), Cu(II), and Zn(II) in model solutions and freshwaters: lability and the d electron configuration. AB - The kinetic speciation of Co(II), Ni(II), Cu(II), and Zn(II) in model solutions of a well-characterized fulvic acid (Laurentian fulvic acid), freshwater samples from the Rideau River (Ottawa, Ontario), and freshwater samples from the Sudbury (Ontario) area were investigated by the competing ligand exchange method using Chelex 100 as the competing ligand and by inductively coupled plasma-mass spectrometry to measure the dissociation kinetics. The metal species were quantitatively characterized by the rate coefficient for the first-order dissociation of metal complex to free metal ion. This technique can be applied to almost all elements and represents an important advance in our ability to investigate the kinetic availability of metal species in the freshwater environment. The order of the lability of the metal complexes, Co(II) > Ni(II) > Cu(II) < Zn(II), follows the reverse order of the ligand field stabilization energy with the exception of Cu(II); the behavior of Cu(II) is also due to the Jahn-Teller effect, which shortens the equatorial bonds and lengthens the axial bonds of a tetragonally distorted Cu(II)-L6 complex. This study has demonstrated a relationship between the lability of metal-DOM complexes of the 3d transition metals in freshwaters and their d electron configuration. This is the first time that the importance of the d electron configuration on the lability of metal complexes in the freshwater environment has been demonstrated. The slow complexation kinetics of both Ni(II) and Cu(II) suggestthatthe usual equilibrium assumption for freshwaters may be invalid. PMID- 12542293 TI - Molecular-scale speciation of Zn and Ni in soil ferromanganese nodules from loess soils of the Mississippi Basin. AB - Determining how environmentally important trace metals are sequestered in soils at the molecular scale is critical to developing a solid scientific basis for maintaining soil quality and formulating effective remediation strategies. The speciation of Zn and Ni in ferromanganese nodules from loess soils of the Mississippi Basin was determined by a synergistic use of three noninvasive synchrotron-based techniques: X-ray microfluorescence (microXRF), X-ray microdiffraction (microXRD), and extended X-ray absorption fine structure spectroscopy (EXAFS). We show that Ni is distributed between goethite (alpha FeOOH) and the manganese oxide lithiophorite, whereas Zn is bound to goethite, lithiophorite, phyllosilicates, and the manganese oxide birnessite. The selective association of Ni with only iron and manganese oxides is an explanation for its higher partitioning in nodules over the soil clay matrix reported from soils worldwide. This could also explain the observed enrichment of Ni in oceanic manganese nodules. The combination of these three techniques provides a new method for determining trace metal speciation in both natural and contaminated environmental materials. PMID- 12542294 TI - "Nonavailable" soil cadmium is bioavailable to snails: evidence from isotopic dilution experiments. AB - Isotopic dilution techniques were initially used to evaluate the bioavailability of trace metals contained in soils to plants, i.e., the phytoavailability. Here, we use for the first time an isotopic technique to evaluate the zooavailability of cadmium (Cd), i.e., the bioavailability to an animal organism. A terrestrial invertebrate, the snail Helix aspersa, was exposed for 14 days to a polluted soil that was spiked with 109Cd. Isotopic composition of snail tissues was then determined, allowing the computing of the L value, which can be considered as a biological estimate of the bioavailable Cd pool in the soil. It showed that the bioavailable pool of Cd to H. aspersa represented 58% of the total soil Cd. The Evalue, a chemical estimate of the soil Cd labile pool, was calculated with short term isotopic exchange kinetics experiments and a time-dependent model describing the isotopic exchange over time. The E(14 days) value indicated that the labile soil Cd pool represented 49.6% of the total soil Cd. The L value was significantly higher than the E value, showing that snails accessed the nonlabile soil Cd pool, generally considered as nonbioavailable. The nonlabile pool contributed for 16% to the total Cd accumulated by snails. These results showed that the uptake of Cd in the nonlabile pool by soil organisms could be important and should be considered in risk assessment procedures for metal polluted soils. PMID- 12542295 TI - Delta13C values of grasses as a novel indicator of pollution by fossil-fuel derived greenhouse gas CO2 in urban areas. AB - A novel fossil fuel pollution indicator based on the 13C/12C isotopic composition of plants has been designed. This bioindicator is a promising tool for future mapping of the sequestration of fossil fuel CO2 into urban vegetation. Theoretically, plants growing in fossil-fuel-CO2-contaminated areas, such as major cities, industrial centers, and highway borders, should assimilate a mixture of global atmospheric CO2 of delta13C value of -8.02 per thousand and of fossil fuel CO2 of average delta13C value of -27.28 per thousand. This isotopic difference should, thus, be recorded in plant carbon. Indeed, this study reveals that grasses growing near a major highway in Paris, France, have strikingly depleted delta13C values, averaging at -35.08 per thousand, versus rural grasses that show an average delta13C value of -30.59 per thousand. A simple mixing model was used to calculate the contributions of fossil-fuel-derived CO2 to the plant tissue. Calculation based on contaminated and noncontaminated isotopic end members shows that urban grasses assimilate up to 29.1% of fossil-fuel-CO2 derived carbon in their tissues. The 13C isotopic composition of grasses thus represents a promising new tool for the study of the impact of fossil fuel CO2 in major cities. PMID- 12542296 TI - Chemical speciation of Zn, Cd, Cu, and Pb in pore waters of agricultural and contaminated soils using Donnan dialysis. AB - Knowledge of trace metal speciation in soil pore waters is important in addressing metal bioavailability and risk assessment of contaminated soils. Numerous analytical methods have been utilized for determining trace metal speciation in aqueous environmental matrixes; however, most of these methods suffer from significant interferences. The Donnan dialysis membrane technique minimizes these interferences and has been used in this study to determine free Zn2+, Cd2+, Cu2+, and Pb2+ activities in pore waters from 15 agricultural and 12 long-term contaminated soils. The soils vary widely in their origin, pH, organic carbon content, and total metal concentrations. Pore water pM2+ activities also covered a wide range and were controlled by soil pH and total metal concentrations. For the agricultural soils, most of the free metal activities were below detection limit, apart from Zn2+ for which the fraction of free Zn2+ in soluble Zn ranged from 2.3 to 87% (mean 43%). Five of the agricultural soils had detectable free Cd2+ with fractions of free metal ranging from 59 to 102% (mean 75%). For the contaminated soils with detectable free metal concentrations, the fraction of free metal as a percentage of soluble metal varied from 9.9 to 97% (mean 50%) for Zn2+, from 22 to 86% (mean 49%) for Cd2+, from 0.4 to 32.1% (mean 5%) for Cu2+, and from 2.9 to 48.8% (mean 20.1%) for Pb2+. For the contaminated soils, the equilibrium speciation programs GEOCHEM and WHAM Model VI provided reasonable estimates of free Zn2+ fractions in comparison to the measured fractions (R2 approximately 0.7), while estimates of free Cd2+ fractions were less agreeable (R2 approximately 0.5). The models generally predicted stronger binding of Cu2+ to DOC and hence lower fractions of free Cu2+ as compared with the observed fractions. The binding of Cu2+ and Pb2+ to DOC predicted by WHAM Model VI was much strongerthan that predicted by GEOCHEM. PMID- 12542297 TI - Reinterpreting literature sorption data considering both absorption into organic carbon and adsorption onto black carbon. AB - We hypothesized that the sorption of polycyclic aromatic hydrocarbons (PAHs) to natural sediments and soils should consider both absorption into a biogenic/diagenetic organic carbon (OC) fraction and adsorption onto a combustion derived, black carbon (BC) fraction. Here, two sets of literature data were reevaluated to illustrate that an OC absorbent and a BC adsorbenttogether can (1) account for sediment--pore-waterdistribution coefficients observed in the field that are greater than predicted by a simple f(OC)K(OC) partitioning model and (2) explain a group of nonlinear phenanthrene isotherms observed in the laboratory with a single value for the BC-normalized distribution coefficient (log K(BC) = 6.1 i 0.04) and a Freundlich exponent (n approximately 0.6 if log K(OC) = 4.0) that is strongly dependent on the K(OC) value selected. PMID- 12542298 TI - Evaluation and testing of analytical methods for cyanide species in municipal and industrial contaminated waters. AB - Total cyanide analysis by distillation is used most commonly to assess cyanide content of water samples. This manual method is robust but slow and provides no information about cyanide speciation, a significant limitation in that cyanide species have substantially different toxicity characteristics. Seven alternative methods for the analysis of cyanide species or groups of species were evaluated in reagent water and five different contaminated water matrices, including five species-specific methods--weak acid dissociable (WAD) cyanide, free cyanide by microdiffusion, available cyanide, automated WAD cyanide by thin film distillation, metal cyanides by ion chromatography--and two automated techniques for total cyanide--total cyanide bythin film distillation and total cyanide by low-power UV digestion. The species-specific cyanide analytical techniques achieved low, ppb-level detection limits and exhibited satisfactory accuracy and precision for most contaminated waters. Analysis of low concentrations of cyanide species in raw wastewater was problematical for the available cyanide and ion chromatography methods, which experienced significant interference problems and/or low recoveries. There was recovery of significant diffusible cyanide in microdiffusion tests with nickel-cyanide-spiked samples, reflecting dissociation of this weak metal-cyanide complex during the test and demonstrating that the test can recover some fraction of WAD cyanide in addition to free cyanide. The automated total cyanide methods, which involve UV digestion, achieved low detection limits for most waters but exhibited low recoveries for some waters. PMID- 12542299 TI - Sorption kinetics and microbial biodegradation activity of hydrophobic chemicals in sewage sludge: model and measurements based on free concentrations. AB - In the current study, a new method is introduced with which the rate-limiting factor of biodegradation processes of hydrophobic chemicals in organic and aqueous systems can be determined. The novelty of this approach lies in the combination of a free concentration-based kinetic model with measurements of both free and total concentrations in time. This model includes microbial biodegradation activity of the chemical in the aqueous phase and chemical sorption kinetics with respect to organic carbon and aqueous phases. The time dependency of free and total concentrations of 7-acetyl-1,1,3,4,4,6 hexamethyltetrahydronaphthalene and 7-acetyl-1,1,3,4,4,6-hexahydro-4,6,6,7,8,8 hexamethylcyclopenta(g)-2-benzopyrane in activated sludge was experimentally determined in vitro. Evaporation losses from the test system were also determined. Least-squares regression to optimize the model parameters resulted in a model that is in accordance with the experimental data. Additionally, the model shows that a comparison between the decrease of free and total chemical concentrations in time, in combination with an independent measurement of the organic carbon/water partition coefficient provides information aboutthe rate limiting step of the degradation process. This information can be used by sewage treatment plant managers to decide whether the microbial biodegradation activity itself or the desorption from organic carbon to the aqueous phase should be improved. PMID- 12542300 TI - Spectroelectrochemical sensing based on multimode selectivity simultaneously achievable in a single device. 11. Design and evaluation of a small portable sensor for the determination of ferrocyanide in Hanford waste samples. AB - A portable spectroelectrochemical sensor has been designed, evaluated, and demonstrated on a complex sample of radioactive waste. The sensor consisted of a black delrin sample compartment with a total internal sample volume of 800 microL, attached to an indium tin oxide coated glass multiple internal reflection optical element. Detection was by total internal reflection of light from a blue light emitting diode source. After a 10 min uptake for each standard, the sensor showed a linear response in absorbance change for 5 x 10(-5) to 5 x 10(-3) M ferrocyanide with electrochemical modulation by scanning at 20 mV/s from -0.30 V to +0.55 V vs a Ag/AgCl reference electrode. Due to the complex nature of Hanford radioactive tank waste samples containing ferrocyanide, a standard addition method was developed for analysis. The spectroelectrochemical sensor determined a concentration of 9.2 mM ferrocyanide for U-Plant-2 simulant solution containing 9.38 mM ferrocyanide that was prepared according to Hanford process flowsheets. A radioactive tank waste sample from Hanford Tank 241-C-112 was determined to be 1.0 mM in ferrocyanide using the spectroelectrochemical sensor. A value for the ferrocyanide concentration in the sample of 0.61 mM was determined by FTIR spectroscopy. PMID- 12542301 TI - Atmospheric pressure mass spectrometry: a new analytical chemical characterization method for dissolved organic matter in rainwater. AB - The complex mixture of organic compounds in the atmosphere influences climate, air quality, and ecosystem processes. Atmospheric pressure electrospray ionization mass spectrometry (APESI-MS) was evaluated as a potential tool for direct measurement of the total suite of individual dissolved organic matter (DOM) compounds in rainwater. The APESI-MS response was linear to all DOM compounds of atmospheric significance examined as standard solutions. Urban precipitation samples from New Brunswick, NJ (USA) were analyzed by APESI-MS over the mass-to-charge (m/z) range 50-3,000. Over 95% of the m/z ions detected were in the low m/z range (50-500). Over 300 unique m/z ions were detected across the 11 rainwater samples indicating the complexity of the mixture of DOM in rainwater. Forty percent of the organic bases (positive mode detection) and 22% of the organic acids (negative mode) occurred in at least 6 of the 11 rainwater samples. Ions corresponding to the m/z of carboxylic acids standards (nonanedioic acid; 1,4-butanedicarboxylic acid; pentanedioic acid; hydroxybutanedioic acid; and butanedioic acid) and to reduced N standards (allylurea; caffeine; imidazole; and N-2-propenylurea) occurred in at least one of the 11 rainwater samples. Total dissolved organic carbon (DOC) estimated from the APESI-MS analysis and measured by standard DOC methods were not statistically different. PMID- 12542302 TI - In situ trace metal speciation in lake surface waters using DGT, dialysis, and filtration. AB - In situ measurements of Fe and Mn by dialysis and diffusive gradients in thin films (DGT) in 5 lakes (pH 4.7-7.5, ionic strength 0.3-5 mmol l(-1)) and Cu and Zn in an acidic and circumneutral lake were compared to results from on site filtration. For the most acidic lake (pH 4.7) all measurements agreed, indicating an absence of colloids and negligible complexation by organic matter. There was little difference in the Mn concentrations measured by the three techniques for any lake, consistent with it being free from complexation. Zn measured by dialysis in circumneutral water was only slightly higher than DGT measurements, appropriate to only partial complexation. Substantial differences between dialysis and DGT for Cu were consistent with complexation by fulvic and humic substances, though not to the extent predicted by the speciation code WHAM. To achieve a good fit it was necessary to adjust the pK for Cu-fulvic binding from 0.8 to 1.3 and to assume that fulvic substances dominated. The presence of low molecular weight strong binding ligands would also be consistent with the data. Differences between the three measurement methods were greatest for Fe, attributable to the presence of large oxyhydroxide colloids, organic complexation and low molecular weight, reactive hydrolysis products. Fe and Mn concentrations measured by DGT on samples returned to the laboratory were much lower than in situ concentrations, illustrating the need for in situ measurements. While use of two in situ techniques provided useful information on the speciation of these natural waters, further refinements are required for unambiguous characterization of the solution. The use of DGT with a more restricted gel that excludes complexes with humic substances should provide complementary information to in situ dialysis. PMID- 12542303 TI - Highly enhanced photoreductive degradation of perchlorinated compounds on dye sensitized metal/TiO2 under visible light. AB - This study reports an example of visible-light photocatalyst based on TiO2 modified by ruthenium-complex sensitizers and noble metal deposits. The photodegradation of trichloroacetate (TCA) and carbon tetrachloride was used as a probe reaction for evaluating the visible light activity of the photocatalyst under the illumination of lambda > 420 nm. Photodeposition of platinum nanoparticles on dye-sensitized TiO2 (Pt/TiO2/Ru(II)L3) drastically enhanced the degradation rate of TCA and CCl4. The visible light reactivity of Pt/TiO2/Ru(II)L3 was optimal with [Ru(II)L3] = 10 microM, [TiO2] = 0.5 g/L, and Pt loading of about 0.2 wt %. Although no electron donors to regenerate the oxidized Ru-sensitizers were added in the aqueous suspension, the photoreductive dechlorination of perchlorinated compounds proceeded far beyond the stoichiometric limit of the initial sensitizer concentration. Water acted as an electron donor to regenerate the sensitizer with a concurrent production of dioxygen. On the other hand, Pt/TiO2/Ru(II)L3 was completely inactive in the presence of dissolved oxygen and the in-situ generated dioxygen gradually decelerated the dechlorination rate. Conduction band electrons transferred to O2 in preference to CCl4 and TCA on Pt deposits. Other noble metals (Ag, Au, and Pd) deposited on TiO2 showed a better oxygen-tolerance but less visible-light reactivity than PtTiO2/Ru(II)L3. Effects of metal loading on the visible light activity and its implications for the efficientvisible-light photocatalyst development are discussed. PMID- 12542304 TI - Design of a photocatalyst for bromate decomposition: surface modification of TiO2 by pseudo-boehmite. AB - The rate of BrO3- reduction by a commercial TiO2 photocatalyst under UV illumination in an aqueous solution was increased by lowering the pH from 7 to 5. The effect is attributable to an enhancement of the electrical interaction between BrO3- and the positively charged surfaces of the TiO2 photocatalyst. The surface charge can be controlled by a surface modification of the TiO2 photocatalyst without controlling the pH of the water. In fact, the isoelectric point of surface-modified TiO2 was higher than that of the unloaded TiO2 photocatalyst, resulting in an increase in the rate of the photocatalytic reduction of BrO3- at a neutral pH. This increase is explained by an increase in the amount of adsorbed BrO3- on the photocatalyst surface. PMID- 12542305 TI - The effect of natrojarosite addition to mine tailings. AB - An increasingly common practice for metallurgical plants is to discard their wastes by combining them with mine tailings prior to disposing the blended material to a containment facility. This practice has occurred since 1985 at the Kidd Creek tailings impoundment where natrojarosite, a waste produced from the adjacent Zn refinery, is combined with mine tailings and is deposited in a single impoundment. To assess the environmental impact of the co-disposal, a laboratory column experiment was conducted. The column material was flotation tailings from the Kidd Creek site containing 3 wt % natrojarosite residue. Dilute sulfuric acid was passed through the column to simulate the acid generated in the unsaturated zone of the tailings impoundment. The results of this experiment were compared to the results of a previous experiment conducted on unamended flotation tailings. The results showed that the effluent from the column containing the natrojarosite bearing mixture had a faster decrease in pH, earlier increases in the concentrations of dissolved metals such as Pb and Cd, and a greater persistence in effluent metal concentrations such as Pb, Zn and Ni. To prevent the observed enhanced release of dissolved metals from mine waste disposal areas, natrojarosite should not be co-disposed with tailings. PMID- 12542306 TI - Recycling of hazardous solid waste material using high-temperature solar process heat. 2. Reactor design and experimentation. AB - A novel high-temperature solar chemical reactor is proposed for the thermal recycling of hazardous solid waste material using concentrated solar power. It features two cavities in series, with the inner one functioning as the solar absorber and the outer one functioning as the reaction chamber. The solar reactor can handle thermochemical processes at temperatures above 1,300 K involving multiphases and controlled atmospheres. It further allows for batch or continuous mode of operation and for easy adjustment of the residence time of the reactants to match the kinetics of the reaction. A 10-kW solar reactor prototype was designed and tested for the carbothermic reduction of electric arc furnace dusts (EAFD). The reactor was subjected to mean solar flux intensities of 2,000 kW m( 2) and operated in both batch and continuous mode within the temperature range of 1,120-1,400 K. Extraction of over 90% of the toxic compounds originally contained in the EAFD was achieved while the condensable products of the off-gas contained mainly Zn, Pb, and Cl. The use of concentrated solar energy as the source of process heat offers the possibility of converting hazardous solid waste material into valuable commodities for processes in closed and sustainable material cycles. PMID- 12542307 TI - Characterization of gamma-alumina-supported manganese oxide as an incineration catalyst for trichloroethylene. AB - Trichloroethylene (TCE) decomposition over a MnOx/ gamma-Al2O3 catalyst in a fixed-bed reactor was conducted in this study. The MnOx/gamma-Al2O3 powders were prepared by the incipient wetness impregnation method with aqueous solution of manganese nitrate. The catalysts were characterized by DTA-TGA, XRD, porosity analysis, SEM, EDX, and XPS. The results show that the main distinct weight loss is found at the temperature around 373 and 873 K,the MnO peaks (2theta = 34.9 degrees and 40.5 degrees) are only observed crystal phase on the fresh catalyst, the SEM image of the MnOx-impregnated gamma-Al2O3 support is much different from the calcined catalyst, and the Mn element quantity on the catalyst surface is higher than that of the impregnated support. The products and reactants distributions from the oxidation of TCE over MnOx/gamma-Al2O3 were analyzed by GC. The results show that the TCE conversion starts from 5% at 443 K and rises to very high values in the 673-873 K ranges and that the CO2 yield also pushes to 99% at the same temperature ranges. HCl and Cl2 are the other main products with little halogenated VOC intermediates. PMID- 12542308 TI - Speciation and transport of heavy metals and macroelements during electroremediation. AB - Electroremediation makes treatment of contaminated clay soils possible. The external electrical field causes several transport processes and changes in soil chemistry. This study concerns the leachability and transport of calcium, magnesium, copper, zinc, lead, nickel manganese, chromium, and iron during treatment with an electric field of soil from a chlor-alkali factory. As expected, most elements were removed from the acidic part of the soil and accumulated in the zone where pH changed from acidic to alkaline. However, acidic leaching of the soil in this zone did not mobilize the elements. Lead formed both an anionic complex which electromigration transported toward the anode as well as a cationic lead fraction which moved toward the cathode. The anionic complex could be lead sulfate. Lead from both fractions was strongly attached to the soil after treatment. The low availability of metals and macroelements after electrokinetic remediation could make electroremediation, excavation, and deposition of the accumulation zone an alternative for the treatment of contaminated soils. PMID- 12542309 TI - Removal of the natural hormone estrone from aqueous solutions using nanofiltration and reverse osmosis. AB - The ability of a variety of nanofiltration and reverse osmosis membranes to retain the natural hormone estrone are examined here as a function of solution conditions. While size exclusion dominates retention with the tighter membranes, both size exclusion and adsorptive effects appear to be instrumental in maintaining high retention on nanofiltration membranes that otherwise exhibit relatively low ion retentions. These adsorptive effects may be driven by hydrogen bonding between estrone and the membrane. Electrostatic attraction appears to aid retention with an apparent slight decrease in retention at high NaCl concentrations. Deprotonation of estrone leads to a significant decrease in retention, most likely as a result of the effect of strong electrostatic repulsive forces decreasing the proximity of the negatively charged estrone to the negatively charged membrane surface and thus lowering the potential for adsorptive retention. This deprotonation effect is absent for tight RO membranes. The results reported here indicate that while open nanofiltration membranes may be effective in retaining estrone under some conditions, the extent of retention may be very susceptible to maintenance of adsorptive capacity at the membrane surface and depend on solution chemistry. PMID- 12542310 TI - Stabilization of available arsenic in highly contaminated mine tailings using iron. AB - To evaluate the stabilization of available As in contaminated tailings from two abandoned metal mines of South Korea (the Myoungbong and Daduck Mines, 6670 and 56,600 mg/kg total As, respectively), characteristics of the tailings were investigated, and the tailings were treated through precipitation of amorphous iron compounds. Steep decreasing trends of extractable (5% NaOCl) As with increasing initial Fe(III) additions were observed in both treated tailings. In general, the treated tailings had the lowest extractable As concentration at pH 6. Available As, defined as the sum of As concentrations for the first four steps of a sequential extraction, was reduced from 2090 to 428 mg/kg (80% reduction) in the Myoungbong tailings and from 1320 to 395 mg/kg (70% reduction) in the Daduck tailings. As levels in the treated tailings decreased even more after a 1-month dormant period. Adsorption/coprecipitation tests performed with mixed As(III) and Fe(III) solutions demonstrated dramatically increased As sequestration via interaction with amorphous iron compounds with increasing pH. The bulk of the As appeared to be affiliated with stable Fe precipitates. PMID- 12542311 TI - Evaluation of the vehicle inspection/maintenance program in the Metropolitan Area of Mexico City. AB - The Inspection/Maintenance Program in the Metropolitan Area of Mexico City (MAMC) mandates a test every 6 months for all gasoline motor vehicles as one of the strategies to decrease emissions of vehicular pollutants. FTP-75 and ASM procedures were performed in our facilities to a fleet of 108 in-use motor vehicles before and after the approval of the I/M mandatory test When our laboratory-simulated ASM data were compared with those of the official certificate, a large difference was observed between them. On the other hand, audits at the test-only centers indicate poor maintenance of the analytical instruments and dynamometers. On the basis of our FTP results, an estimation of the emissions change for the MAMC fleet shows a net 4% decrease in CO emissions, while total hydrocarbons and NOx increased 9 and 8%, respectively. Our results indicate that the I/M system in the MAMC lacks the technical capability and investment to ensure that software and hardware are properly maintained, calibrated, and upgraded. Sometimes limited attention is paid to ensure adequate training of inspectors, auditors, and quality control staff. PMID- 12542312 TI - Wide-field compensation of monochromatic eye aberrations: expected performance and design trade-offs. AB - The optical quality of the human eye varies across the visual field. Hence an exact compensation of the eye aberration for a given field point can give rise to a less-than-optimum compensation in neighboring field regions. We have studied some aspects of this problem and present here an approach to design wide-field (< 10 degrees) optically thin correcting elements, e.g., phase plates, deformable mirrors, and liquid-crystal displays. Their expected performance is assessed using actual eye aberration data. Particular attention is given to the design of elements providing a minimum averaged rms residual aberration and those providing a nearly uniform rms residual aberration across a given field. PMID- 12542313 TI - Foveal contrast thresholds exhibit spatial-frequency- and polarity-specific contour interactions. AB - Traditionally, contour interaction has been investigated at the visual acuity limit using a Landolt C and flanking bars, performance being quantified in terms of a percent correct measure. More recently, it has been shown that the properties of the contour interaction are different when larger stimuli are used: Contour interaction is not polarity specific, and spatial frequency tuning for an unflanked C is broader. Here we quantify contour interaction for stimuli 5x larger than the resolution limit in terms of contrast thresholds. We show that polarity of bars has little effect on unfiltered stimuli but does show very different effects on the spatial-frequency-tuning curves for discrimination of the Landolt C. This explains the polarity dependence of crowding at the visual acuity limit and its independence for larger unfiltered targets. Thus the underlying filtering function is composed of more than one mechanism, affected differently depending on the relative polarity of the test and flank contours. PMID- 12542314 TI - Wavelength dependence of reflectometric cone photoreceptor directionality. AB - We present evidence for the wavelength dependence of the directionality of light reflected from cone receptor cells (optical Stiles-Crawford effect): Blue light is more directional than red. According to the waveguide-scattering model of Marcos et al. [J. Opt. Soc. Am. A 15, 2012 (1998)], directionality is the sum of a waveguide component and a scattering component. The latter is proportional to 1 over wavelength squared, and it is related to the row-to-row spacing of the cone lattice. Our results allow a firm confirmation of Marcos et al.'s theory. For a 1.9-deg foveal area, group mean (n = 18) cone spacing was 3.42 microm, in good agreement with anatomical data. Group mean waveguide directionality was 0.077 mm( 2). PMID- 12542315 TI - The KGBR viewpoint-lighting ambiguity. AB - We address the visual ambiguities that arise in estimating object and scene structure from a set of images when the viewpoint and lighting are unknown. We obtain a novel viewpoint-lighting ambiguity called the KGBR that corresponds to a group of three-dimensional affine transformations on the object or scene geometry combined with transformations on the object or scene albedo. Our analysis assumes orthographic projection with an affine camera model. We include photometric cues, such as shadowing and shading, that we model using Lambertian reflectance functions with shadows (cast and attached) and multiple light sources (but no interreflections). We relate the KGBR to affine ambiguities in estimating shape and to the generalized bas-relief (GBR) ambiguity. PMID- 12542316 TI - Statistical-physical model for foliage clutter in ultra-wideband synthetic aperture radar images. AB - Analyzing foliage-penetrating (FOPEN) ultra-wideband synthetic aperture radar (SAR) images is a challenging problem owing to the noisy and impulsive nature of foliage clutter. Indeed, many target-detection algorithms for FOPEN SAR data are characterized by high false-alarm rates. In this work, a statistical-physical model for foliage clutter is proposed that explains the presence of outliers in the data and suggests the use of symmetric alpha-stable (SalphaS) distributions for accurate clutter modeling. Furthermore, with the use of general assumptions of the noise sources and propagation conditions, the proposed model relates the parameters of the SalphaS model to physical parameters such as the attenuation coefficient and foliage density. PMID- 12542317 TI - Phase retrieval by iterated projections. AB - Several strategies in phase retrieval are unified by an iterative "difference map" constructed from a pair of elementary projections and three real parameters. For the standard application in optics, where the two projections implement Fourier modulus and object support constraints, respectively, the difference map reproduces the "hybrid" form of Fienup's input-output map when a particular choice is made for two of the parameters. The geometric construction of the difference map illuminates the distinction between its fixed points and the recovered object, as well as the mechanism whereby the form of stagnation encountered by alternating projection schemes is avoided. When support constraints are replaced by object histogram or atomicity constraints, the difference map lends itself to crystallographic phase retrieval. Numerical experiments with synthetic data suggest that structures with hundreds of atoms can be solved. PMID- 12542318 TI - Local correlations, information redundancy, and sufficient pixel depth in natural images. AB - A mathematical framework that permits the factorization of a joint probability distribution into its localized components for a two-dimensional array of pixels is presented. The factorization was used to estimate the contribution to mutual information of two- (I2) and three-pixel (I3) luminance correlations for a large ensemble of natural images analyzed at various spatial scales and pixel depths b. It is shown that both I2 and I3 saturate at b approximately 6 bits per pixel. Three-pixel correlations are shown to produce only a marginal increase of information redundancy (4%) over two-pixel correlations (50%). Implications for neural representation in visual cortex are discussed. PMID- 12542319 TI - Efficient frequency-domain sample selection for recovering limited-support images. AB - An image whose region of support is smaller than its bounding rectangle can, in principle, be reconstructed from a subset of the Nyquist samples. However, determining such a sampling set that gives a stable reconstruction is a difficult and computationally intensive problem. An algorithm is developed for determining periodic nonuniform sampling patterns that is orders of magnitude faster than existing algorithms. The speedup is achieved by using a sequential selection algorithm and heuristic metrics for the quality of sampling sets that are fast to compute, as opposed to the more rigorous linear algebraic metrics that have been used previously. Simulations show that the sampling sets determined using the new algorithm give image reconstructions that are of accuracy comparable with those determined by other slower algorithms. PMID- 12542320 TI - Coherent-mode decomposition of partially polarized, partially coherent sources. AB - It is shown that any partially polarized, partially coherent source can be expressed in terms of a suitable superposition of transverse coherent modes with orthogonal polarization states. Such modes are determined through the solution of a system of two coupled integral equations. An example, for which the modal decomposition is obtained in closed form in terms of fully linearly polarized Hermite Gaussian modes, is given. PMID- 12542321 TI - Novel electromagnetic approach to photonic crystals with use of the C method. AB - We introduce a new method allowing rigorous electromagnetic analysis of scattering through photonic crystals comprising polygonal or round rods. For this purpose, we reformulate the C method with adaptive spatial resolution by utilizing the hybrid-spectrum connection method, permitting the use of nonidentical trapezoidal profiles. Considering polygonal rods as gratings consisting of different piecewise-differentiable surfaces, we are able to analyze the reflection and the transmittance of crystals by means of the C method. To enhance computational efficiency, we apply the recursive S-matrix approach with Redheffer's star product to solve the transfer matrix for structures of numerous successive layers of rods. PMID- 12542322 TI - Light propagation in biological tissue. AB - Biological tissue scatters light mainly in the forward direction where the scattering phase function has a narrow peak. This peak makes it difficult to solve the radiative transport equation. However, it is just for forward-peaked scattering that the Fokker-Planck equation provides a good approximation, and it is easier to solve than the transport equation. Furthermore, the modification of the Fokker-Planck equation by Leakeas and Larsen provides an even better approximation and is also easier to solve. We demonstrate the accuracy of these two approximations by solving the problem of reflection and transmission of a plane wave normally incident on a slab composed of a uniform scattering medium. PMID- 12542323 TI - Light transport in chiral and magnetochiral random media. AB - We present a microscopic approach to study electromagnetic wave propagation in media with broken mirror symmetry. We introduce and calculate the transport mean free path l*C associated with the residual polarization of diffuse light in chiral systems. In chiral media subject to an external magnetic field B, all symmetry requirements exist to create a macroscopic "super" light current in the direction of B that persists even in the absence of a spatial photon density gradient. However, we show that such a current is identically zero in our model. We finally show the existence of a linear magnetotransmission in magnetochiral media. PMID- 12542324 TI - Optimized two-frequency phase-measuring-profilometry light-sensor temporal-noise sensitivity. AB - Temporal frame-to-frame noise in multipattern structured light projection can significantly corrupt depth measurement repeatability. We present a rigorous stochastic analysis of phase-measuring-profilometry temporal noise as a function of the pattern parameters and the reconstruction coefficients. The analysis is used to optimize the two-frequency phase measurement technique. In phase measuring profilometry, a sequence of phase-shifted sine-wave patterns is projected onto a surface. In two-frequency phase measurement, two sets of pattern sequences are used. The first, low-frequency set establishes a nonambiguous depth estimate, and the second, high-frequency set is unwrapped, based on the low frequency estimate, to obtain an accurate depth estimate. If the second frequency is too low, then depth error is caused directly by temporal noise in the phase measurement. If the second frequency is too high, temporal noise triggers ambiguous unwrapping, resulting in depth measurement error. We present a solution for finding the second frequency, where intensity noise variance is at its minimum. PMID- 12542325 TI - Design of curved waveguides: the matched bend. AB - A new criterion for the optimum design of curved dielectric waveguides is proposed. The bends designed according to this model are named matched bends. In the matched bend, the suitable choice of both bending radius and bending angle reduces the total losses of the bend and avoids the leaky-mode excitation at the end of the bend. For a given angle, a discrete number of bending radii that satisfy the matched bend criterion can be analytically determined. With respect to the lateral offset, matched bends are more robust to both fabrication tolerances and wavelength and can be realized in every technology. The reduction of the leaky-mode excitation at the output of the bend is a fundamental property when two or more components are cascaded. Ghost images in the spectral response, cross talk, and asymmetries of the transfer function are successfully reduced. Some examples that use buried, rib, ridge, and diffused waveguides are presented and discussed. PMID- 12542326 TI - Sensitivity analysis of narrowband photonic crystal filters and waveguides to structure variations and inaccuracy. AB - Photonic crystal microcavities, formed by local defects within an otherwise perfectly periodic structure, can be used as narrowband optical resonators and filters. The coupled-cavity waveguide (CCW) is a linear array of equally spaced identical microcavities. Tunneling of light between microcavities forms a guiding effect, with a central frequency and bandwidth controlled by the local defects' parameters and spacing, respectively. We employ cavity perturbation theory to investigate the sensitivity of microcavities and CCWs to random structure inaccuracies. For the microcavity, we predict a frequency shift that is due to random changes in the lattice structure and show an approximate linear dependence between the standard deviation of the structure inaccuracy and that of the resonant frequency. The effect of structural inaccuracy on the CCW devices, however, is different; it has practically no effect on the CCW performance if it is below a certain threshold but may destroy the CCW if this threshold is exceeded. PMID- 12542327 TI - Group velocity and dispersion model of coupled-cavity waveguides in photonic crystals. AB - A theoretical model of the group velocity, dispersion parameter, and dispersion slope of coupled-cavity waveguides in photonic crystals is reported. Results arising from closed-form expressions show a good agreement with simulation results obtained by employing a plane-wave expansion method. Coupled-cavity waveguides present interesting dispersion properties that may be employed in applications such as optical signal processing, dispersion compensation, and optical delay lines. PMID- 12542328 TI - Resonator stability subject to dynamic random-tilt aberration. AB - We derive the behavior of the average exit time (i.e., the number of reflections before escape) of a ray path traveling between two perfect mirrors subject to dynamic random-tilt aberrations. Our calculation is performed in the paraxial approximation. When small random tilts are taken into account, we may consider an asymptotic regime that generically reduces the problem to the study of the exit time from an interval for a harmonic, frictionless oscillator driven by Gaussian white noise. Despite its apparent simplicity, the exact solution of this problem remains an open mathematical challenge, and we propose here a simple approximation scheme. For flat mirrors, the natural frequency of the oscillator vanishes, and, in this case, the average exit time is known exactly. It exhibits a 2/3 scaling-law behavior in terms of the variance of the random tilts. This behavior also follows from our approximation scheme, which establishes the consistency of the scaling law. Our mathematical results are confirmed with simulation experiments. PMID- 12542329 TI - Dispersion compensation in whispering-gallery modes. AB - We show that manipulation by a spatial profile of the refractive index of a circularly symmetric dielectric cavity results in a novel way of fine tuning frequency separations as well as spatial localizations of high-Q whispering gallery modes excited in the cavity. The method permits dispersion compensation in the modes (spectrum equalization), diminishes the quality-factor limitation by surface roughness and contamination, and allows critical coupling to ultra-high-Q modes without maintaining an air gap with evanescent couplers. PMID- 12542330 TI - Circularly polarized beams and vortex generation in uniaxial media. AB - We deduce the expressions for the two circularly polarized components of a paraxial beam propagating along the optical axis of a uniaxial crystal. We find that each of them is the sum of two contributions, the first being a free field and the second describing the interaction with the opposite component. Moreover, we expand both components as a superposition of vortices of any order, thus obtaining a complete physical picture of the interaction dynamics. Consequently, we argue that a left-hand circularly polarized incoming beam, endowed with a circular symmetric profile, gives rise, inside the crystal, to a right-hand circularly polarized vortex of order 2. The efficiency of this vortex generation is investigated by means of a power exchange analysis. The Gaussian case is fully discussed, showing the relevant features of the vortex generation. PMID- 12542331 TI - Transmission and reflection of Gaussian beams by anisotropic parallel plates. AB - Explicit and compact expressions describing the reflection and the transmission of a Gaussian beam by anisotropic parallel plates are given. Multiple reflections inside the plate are taken into account as well as arbitrary optical axis orientation and angle of incidence. PMID- 12542332 TI - Remote sensing of penetrable objects buried beneath two-dimensional random rough surfaces by use of the Mueller matrix elements. AB - The modified Mueller matrix elements for electromagnetic scattering from penetrable objects buried under two-dimensional random rough surfaces are investigated. This matrix relates the incident to the scattered waves, and it contains different combinations of the fully polarimetric scattering matrix elements. The statistical average of each Mueller matrix element is computed on the basis of the Monte Carlo simulations by exploiting the speed of the three dimensional steepest-descent fast multipole method. The numerical results clearly show that relying only on the co-polarized or the cross-polarized intensities or both (i.e., vv, hh, vh, and hv) is not sufficient for sensing the buried objects. However, examining all 16 Mueller matrix elements significantly increases the possibility of detecting these objects. This technique can be used in remote sensing of scatterers buried beneath the rough ground. PMID- 12542334 TI - Puupehenone congeners from an indo-pacific hyrtios sponge. AB - An investigation of the constituents from an Indonesian Hyrtios sponge has provided new insights about the chemistry and biology of the puupehenones, a unique class of merosesquiterpenes. The parent compound, puupehenone (2), has been repeatedly encountered in sponges from four distinct orders. In this study we characterized three compounds, (+)-(5S,8S,9R,10S)-20-methoxypuupehenone (3), (+)-(5S,8S,10S)-20-methoxy-9,15-ene-puupehenol (4), and (+)-(5S,8S,9R,10S)-15,20 dimethoxypuupehenol (5). Their structures were supported by complete sets of spectroscopic data along with comparisons to literature properties. While 5 was observed in the crude extracts, it was also heat labile and could be converted at 35 degrees C to a mixture of 3 and 4. The possibility that 3, 4, and 5 are formed from 2 by a series of methanol additions is discussed. The bioactivity of these compounds in soft-agar cytotoxicity tests was also explored. PMID- 12542335 TI - Characterization of a rhamnogalacturonan and a xyloglucan from neriumindicum and their activities on PC12 pheochromocytoma cells. AB - Two polysaccharides, a rhamnogalacturonan (J1) and a xyloglucan (J2), were isolated and purified from the whole flowers of Nerium indicum and characterized by GC, GC-MS, NMR, and ESMS. The rhamnogalacturonan was found to consist of repeating units of a linear trisaccharide with the structure [-->4)-alpha-GalpA(1 ->2)alpha-Rhap(1-->4)-alpha-GalpA(1-->4)-](n)(), while the xyloglucan consisted of repeating units of the trisaccharide, which was (1-->4)-linked beta glucopyranosyl residues as a backbone and substituted at position 6 linked to terminal alpha-xylopyranose side chains. These two polysaccharides were tested on the proliferation and differentiation on PC12 pheochromocytoma cells and found to have effects similar to nerve growth factor (NGF). PMID- 12542336 TI - New bisindole alkaloids from Tabernaemontana corymbosa. AB - Ten new bisindole alkaloids of the vobasinyl-ibogan type, viz., conodiparines A-F (1-6), conodutarines A and B (7, 8), and cononitarines A and B (9, 10), were obtained from the leaf extract of the Malayan species Tabernaemontana corymbosa. The structures were determined using NMR and MS analysis. PMID- 12542337 TI - Absolute configuration of a tetrahydrophenanthrene from Heliotropium ovalifolium by LC-NMR of its Mosher esters. AB - A new tetrahydrophenanthrene (1, (1R,2R)-1-hydroxy-2-methoxy-6,9-dimethyl-2,3 dihydrophenanthren-4(1H)-one (heliophenanthrone)) has been isolated from the aerial parts of Heliotropium ovalifolium. Its structure was elucidated on the basis of spectroscopic data, and the absolute configuration of the asymmetric centers was determined from LC-NMR data of the Mosher ester derivatives. PMID- 12542338 TI - Acetylene sesquiterpenoid esters from the green alga Caulerpa prolifera. AB - Sixteen secondary metabolites of the green alga Caulerpa prolifera have been isolated, and their chemical structures elucidated by analysis of their spectroscopic data. Two groups of metabolites have been established, with either a 1,2-dihydro- (2a-2i) or a 1,2,3,3'-tetrahydro-2,3-didehydro (3a-3f) caulerpenyne carbon backbone. The terminal vinyl acetoxy group of caulerpenyne was substituted by various fatty acid residues. The antifouling activity of the algal extract was tested in laboratory assays against two of the major groups of fouling organisms (bacteria, microalgae). PMID- 12542339 TI - Yardenone A and B: new cytotoxic triterpenes from the Indian Ocean sponge Axinella cf. bidderi. AB - Two new polyepoxysqualene-derived triterpenes, yardenone A (1) and B (2), together with the known yardenone (3) and sodwanone A (4), have been isolated from the marine sponge Axinella cf. bidderi from Yemen's Socotra Island in the Indian Ocean. The structures were elucidated using spectroscopic data. The relative stereochemistry was established by the analysis of ROESY spectra as well as coupling constants and molecular modeling. Furthermore, the absolute configuration of 1 was confirmed by the advanced Mosher's method. The cytotoxicity of these compounds was evaluated against a NSCLC cell line. PMID- 12542340 TI - Diterpenes from Colophospermum mopane: "missing links" in the biogenesis of 9,13 epoxylabdanes. AB - Moponeol A (1) and moponeol B (2) were isolated from Colophospermum mopane along with a mixture of their corresponding aldehydes (3 and 4). These substances are primitive diterpenes that we view as the "missing links" in the biosynthesis of the 9,13-epoxylabdanes. The structures of 1 and 2 were elucidated by a combination of spectra (NMR and MS) of the isolates and their mono-p-bromobenzoyl derivatives. The structures of 3 and 4 were confirmed by their ready reduction to 1 and 2. The biosynthetic implications of the stereochemical assignments of these terpenoids are briefly discussed. PMID- 12542341 TI - New terpene hydrocarbons from the alligatoridae (crocodylia, reptilia). AB - The contents of the paracloacal gland secretions of the alligatorids Alligator mississippiensis, A. sinensis, Paleosuchus palpebrosus, and P. trigonatus were investigated. Novel acyclic hydrocarbon terpenes with a rare trisubstituted 2,4 diene system were identified in the secretions of A. sinensis, P. palpebrosus, and P. trigonatus. The structures of the monoterpene (2E,4E)-3,7-dimethyl-2,4 octadiene (9) and the sesquiterpene (2E,4E,7S)-3,7,11-trimethyl-2,4-dodecadiene (14) were proven by synthesis and gas chromatography on a chiral phase. Several other new terpenes (11, 16, 17, 18, 19, and 20) related to these components also were present in the secretions, as well as the known compounds myrcene (6), (E) beta-farnesene (4), (E)-beta-springene (3), squalene (5), cembrene A (1), and 11,12-dihydrocembren-10-one (2). PMID- 12542342 TI - Flavanone glycosides from Miconia trailii. AB - Assay-guided fractionation of the ethanol extract of the twigs and leaves of Miconia trailii yielded two new flavanone glycosides, matteucinol 7-O-alpha-l arabinopyranosyl(1-->6)-beta-d-glucopyranoside (miconioside A, 1) and farrerol 7 O-beta-d-apiofuranosyl(1-->6)-beta-d-glucopyranoside (miconioside B, 2), along with the known compounds matteucinol 7-O-beta-d-apiofuranosyl(1-->6)-beta-d glucopyranoside (3), matteucinol (4), 2alpha,3beta,19alpha-trihydroxyolean-12-ene 24,28-dioic acid (bartogenic acid, 5), 2alpha,3beta,23-trihydroxyolean-12-ene-28 oic acid (arjunolic acid, 6), 2alpha,3alpha,19alpha, 23-tetrahydroxyurs-12-ene-28 oic acid (myrianthic acid, 7), and stigmast-4-ene-3,6-dione (8). The structures of 1-8 were elucidated by spectroscopic methods, including 2D NMR. PMID- 12542344 TI - A new sesquiterpenoid quinone and other related compounds from the Okinawan marine sponge Dactylospongia elegans. AB - The new sesquiterpenoid quinone, neodactyloquinone (1), and dactylolactones A-D (2-5) were isolated from the Okinawan sponge Dactylospongia elegans. The structures of these compounds were determined by spectroscopic analysis. PMID- 12542343 TI - PSII inhibitory activity of resorcinolic lipids from Sorghum bicolor. AB - Resorcinolic lipids were isolated from the root extracts of Sorghum bicolor and identified as 4,6-dimethoxy-2-[(8'Z,11'Z)-8',11',14'-pentadecatriene]resorcinol (4), 4-methoxy-6-ethoxy-2-[(8'Z,11'Z)-8',11',14'-pentadecatriene]resorcinol (5), and 4-hydroxy-6-ethoxy-2-[(10'Z,13'Z)-10',13',16'-heptadecatriene]resorcinol (6). Compounds 4 and 5 inhibited photosynthetic oxygen evolution (IC50 0.09 and 0.20 microM, respectively). Compound 4 could not be enzymatically converted to a quinone, suggesting that the quinone moiety is not required for its photosystem II inhibitory activity. Compounds 5 and 6 are reported for the first time. PMID- 12542345 TI - Melophlins C-O, thirteen novel tetramic acids from the marine sponge Melophlus sarassinorum. AB - Thirteen new metabolites, melophlins C-O (1-13), were identified from the marine sponge Melophlus sarassinorum. Compounds 1-13 represent tetramic acid derivatives that differ with regard to the nature of their alkyl side chains. The structures of the new compounds were elucidated on the basis of comprehensive spectral analysis (1H, 13C, 1H-1H COSY, HMQC, and HMBC NMR, as well as low- and high resolution ESIMS and EIMS). The absolute configurations of 1, 8, 10, 11, 12, and 13 were determined by ESI LC/MS analysis of chiral derivatives obtained upon oxidation and hydrolysis of the respective parent compounds. Melophlin C (1) displayed pronounced antibacterial activity against Bacillus subtilisand Staphylococcus aureus, together with antifungal activity against Candida albicans. PMID- 12542346 TI - Novel chromone derivatives from the fungus Aspergillus versicolor isolated from the marine sponge Xestospongia exigua. AB - From the marine sponge Xestospongia exigua collected in Indonesia the fungus Aspergillus versicolor was isolated. Following cultivation in a seawater-based medium seven new angular tricyclic chromone derivatives (1-7) were obtained from the mycelia and culture filtrate. Compounds 2-7 contain an additional dihydropyran ring system which is replaced by a pyridine ring in 1. The structures of the new natural products were established on the basis of extensive one- and two-dimensional NMR spectroscopic studies (1H, 13C, COSY, HMQC, HMBC, NOE difference spectra) as well as on mass spectral analysis. PMID- 12542347 TI - Metabolites of a blocked chloramphenicol producer. AB - Addition of p-aminophenylalanine (4), an advanced biosynthetic precursor of the antibiotic chloramphenicol (5), to a Streptomyces venezuelae pabAB mutant (VS629) restored chloramphenicol production and led to formation of the non-chlorinated analogue corynecin II (6) and four acetanilide derivatives: p (acetylamino)phenylalanine (7), p-(acetylamino)benzyl alcohol (13), p (acetylamino)benzoic acid (14), and p-(acetylamino)phenol (acetaminophen, 16). Metabolite structures were deduced from NMR and MS-MS data and established by chromatographic and spectroscopic comparisons with authentic samples. Reference compound 13 was synthesized by reducing the acid chloride of 14. Shunt pathways are proposed to account for the formation of the metabolites from p aminophenylalanine. PMID- 12542348 TI - An efficient conversion of (3R,3'R,6'R)-lutein to (3R,3'S,6'R)-lutein (3' epilutein) and (3R,3'R)-zeaxanthin. AB - Two dietary carotenoids, (3R,3'R,6'R)-lutein (1) and (3R,3'R)-zeaxanthin (2), and their metabolite (3R,3'S,6'R)-lutein (3'-epilutein) (3) accumulate in human serum, milk, and ocular tissues. There is increasing evidence that compounds 1 and 2 play an important role in the prevention of age-related macular degeneration. Therefore, the availability of these carotenoids for metabolic studies and clinical trials is essential. Compound 1 is isolated from extracts of marigold flowers (Tagete erecta) and is commercially available, whereas 2 is only accessible by a lengthy total synthesis, and a viable method for synthesis of 3 has not yet been developed. This report describes an efficient conversion of technical grade 1 to 2 via 3. Acid-catalyzed epimerization of 1 yields an equimolar mixture of diastereomers 1 and 3. The mixture was separated by enzyme mediated acylation with lipase AK from Pseudomonas fluorescens that preferentially esterified 3 and after alkaline hydrolysis yielded this carotenoid in 90% diastereomeric excess (de). Compound 3 was also separated from 1 in 56-88% de by solvent extraction and low-temperature crystallization, Soxhlet extraction, or supercritical fluid extraction. Base-catalyzed isomerization of 3 gave 2 in excellent yield, providing a convenient alternative to the total synthesis of this important dietary carotenoid. PMID- 12542349 TI - New bioactive rosigenin analogues and aromatic polyketide metabolites from the freshwater aquatic fungus Massarina tunicata. AB - Four new rosigenin analogues (massarigenins A-D; 1-4) and two new aromatic polyketide-derived secondary metabolites (massarinins A and B; 6, 7) and have been isolated from the freshwater aquatic fungus Massarina tunicata. The structures of these compounds were determined primarily by analysis of NMR data, and that of compound 1 was verified by X-ray crystallography. The known compound 4-(2-hydroxybutynoxy)benzoic acid (11) was also obtained, and its absolute stereochemistry was assigned. Several of these metabolites showed antibiotic activity against Gram-positive bacteria. PMID- 12542350 TI - Cyclorocaglamide, the first bridged cyclopentatetrahydrobenzofuran, and a related "open chain" rocaglamide derivative from Aglaia oligophylla. AB - Two rocaglamide-related natural products, the previously known compound 6 demethoxy-10-hydroxy-11-methoxy-6,7-methylendioxyrocaglamide (3), and cyclorocaglamide (4), its 8b,10-anhydro analogue, have been isolated from the tropical plant Aglaia oligophylla. Compound 4 is the first bridged cyclopentatetrahydrobenzofuran natural product, and it exhibited a CD spectrum virtually opposite that of all the other rocaglamide natural products known so far, but it still has the same absolute configuration at all stereogenic centers of the basic molecular framework. This was shown unequivocally by quantum chemical CD calculations (here based on molecular dynamics-weighted force field structures) and was finally confirmed experimentally, by a "biomimetic-type" cyclization of 3 to give 4, with the expected "inversion" of the CD spectrum. The opposite chiroptical properties of 3 and 4, despite their homochiral character, underline the necessity of handling chiroptical data with the greatest care, e.g., by simulating them by quantum chemical CD calculations. Compound 3 exhibited an LC(50) of 2.5 ppm when evaluated against neonate larvae of Spodoptera littoralis, while 4 was inactive in this assay up to 100 ppm. PMID- 12542351 TI - Structures of new cyclic diarylheptanoids and inhibitors of nitric oxide production from Japanese folk medicine Acer nikoense. AB - Three new cyclic diarylheptanoids, acerosides B1 and B2 and aceroketoside, were isolated together with 20 known compounds from a Japanese folk medicine, the stem bark of Acer nikoense. The absolute stereostructures of the new compounds were determined on the basis of chemical and physicochemical evidence. In addition, the principal diarylheptanoid constituents were found to exhibit inhibitory activity on nitric oxide production in lipopolysaccharide-activated macrophages. PMID- 12542352 TI - Antineoplastic agents 500. Narcistatin. AB - An efficient procedure was found for synthetic conversion of the sparingly soluble anticancer isocarbostyril narciclasine (1), a component of various Narcissus species, to a cyclic phosphate designated narcistatin (3b). The reaction between narciclasine, tetrabutylammonium dihydrogen phosphate, and p toluenesulfonic acid in pyridine afforded pyridinium narcistatin (3a) in reasonable yields. Transformation of narcistatin (3a) to, for example, the water soluble prodrug sodium narcistatin (3d) was easily achieved by cation exchange chromatography. Narcistatin (3b) and 15 salt derivatives were evaluated against a panel of human cancer cell lines, and the range (0.1-0.01) of GI(50) values in micro g/mL was found to parallel that shown by the parent narciclasine. PMID- 12542353 TI - Complement-inhibiting iridoids from Morinda morindoides. AB - Morinda morindoides (Baker) Milne-Redhead (syn. Gaertnera morindoides Bak.) is one of the most popular medicinal plants in the Democratic Republic of Congo. In relation to its traditional use against rheumatic pains, fractionation of both the EtOAc- and the n-BuOH-soluble fraction of the 80% MeOH extract of the leaves, guided by the anticomplementary activity on the classical activation pathway, yielded eight novel iridoids (1-8), all containing a spirolactone functionality. Their structure was elucidated using spectroscopic methods. Gaertneroside 1, acetylgaertneroside 2, and gaertneric acid 5 were found to inhibit the activation of the classical pathway of the complement system, with IC(50) values between 58 and 69 microM. In addition to the biologically active flavonoids reported before from the same plant, these complement-inhibiting iridoids may contribute at least in part to the traditional use against rheumatic pains. PMID- 12542354 TI - Chemical and biological investigation of the fungus Pulveroboletus ravenelii. AB - Two new compounds, pulveraven A (1) and pulveraven B (2), as well as vulpinic acid (3) and its previously unreported polymorph were isolated from the fruiting body of Pulveroboletus ravenelii. The structures were determined using a combination of NMR, MS, IR, optical rotation, molecular modeling, and X-ray analysis. The isolates were evaluated for antimicrobial activity as well as their potential to inhibit cyclooxygenase (COX) activity and carcinogen-induced preneoplastic lesion formation with mouse mammary organ culture (MMOC). PMID- 12542355 TI - New bis-secolabdane diterpenoids from Excoecaria agallocha. AB - Two new bis-secolabdane diterpenoids, excoecarins R1 (1) and R2 (2), were isolated from the resinous wood of Excoecaria agallocha. The structures of 1 and 2 were established on the basis of spectroscopic data interpretation and chemical evidence. PMID- 12542356 TI - Cationic reduction of bastadin-4 to bastadin-5. Preparation of 5-[2h]-bastadin-5 by site-specific isotopic labeling. AB - A chemoselective conversion of bastadin-4 to the important Ca2+ channel modulator bastadin-5 (1a) has been achieved using cationic hydrogenation (Et3SiH, TFA, 60%). Specifically deuterated bastadin-5 (1b, >95 at. %) was prepared following this method and the simplified 1H NMR H-5/H2-6 spin system of 1b exploited to study temperature-dependent macrocyclic ring dynamics. PMID- 12542357 TI - Lakshminine, a new rare oxoisoaporphine alkaloid from Sciadotenia toxifera, and structural revisions of telazoline and teladiazoline, two related oxoaporphines from Telitoxicum peruvianum and T. glaziovii. AB - Lakshminine (1), a novel oxoisoaporphine alkaloid possessing a C-6 amine substituent, was isolated from a basic fraction from the woody vines (collected from two bush-ropes) of Sciadotenia toxifera. This compound represents the first documented occurrence of an oxoisoaporphine from any Menispermaceae species other than Menispermum dauricum. The structures of two related aporphine alkaloids, telazoline (3) and teladiazoline (5), were revised on the basis of a comparison of their spectral data with that of lakshminine (1). PMID- 12542358 TI - iso-caracurine V, a novel unexpected decomposition product of caracurine V. AB - The structure of iso-caracurine V (3), an unexpected decomposition product of the Strychnos alkaloid caracurine V (1), was determined by NMR spectroscopy. Compound 3, which was probably previously regarded as bisnortoxiferine I (2), is a one sided ring closed product of 2. PMID- 12542359 TI - Nodulisporic acids C, C1, and C2: a series of D-ring-opened nodulisporic acids from the fungus Nodulisporium sp. AB - Nodulisporic acids C (4a), C1 (5a), and C2 (6a), a series of D-ring-opened nodulisporic acids, were isolated from fermentations of a mutant culture of Nodulisporium sp. MF5954. Nodulisporic acid C, the most potent of the three, showed good activity against fleas with an LD90 of 10 micro g/mL. These compounds tend to be unstable in the free acid form and were isolated as stable sodium salts. PMID- 12542360 TI - Antitumor agents. 221.1 buceracidins A and B, two new flavanones from Bucida buceras. AB - As part of a study on antitumor agents from rainforest plants, two new flavanones, buceracidin A (1), 5,2'-dihydroxy-3-methoxy-6,7-(2' ',2' ' dimethylchromene)flavanone, and buceracidin B (2), 5,2'-dihydroxy-6,7-(2' ',2' ' dimethylchromene)flavanone, were isolated together with four known prenylated flavanones, minimiflorin (3), 3-hydroxyminimiflorin (4), 3-methoxyminimiflorin (5), and mundulinol (6), from Bucida buceras. The structures of buceracidins A and B were elucidated from detailed 2D NMR analyses. Among the six flavanones, 3, 5, and 6 were marginally cytotoxic in a human tumor cell line panel. PMID- 12542361 TI - New icetexane and 20 norabietane diterpenes with trypanocidal activity from Dracocephalum komarovi. AB - Two new icetexane diterpenes, cyclocoulterone (1) and komaroviquinone (2), and a novel 20-norabietane diterpene, dracocephalone A (3), were isolated from Dracocephalum komarovi. Their structures were elucidated by extensive analyses of spectral data. Komaroviquinone (2) showed strong trypanocidal activity against epmastigotes of Trypanosoma cruzi, the causative agent of American trypanosomiasis, with a minimum lethal concentration of 0.4 micro M. PMID- 12542362 TI - Three new malyngamides from a Papua New Guinea collection of the marine cyanobacterium Lyngbya majuscula. AB - Three new malyngamides, U (1), V (2), and W (3), all possessing an uncommon oxygenation pattern, have been isolated from the marine cyanobacterium Lyngbya majuscula collected in Papua New Guinea. The planar structures of these compounds were deduced from NMR data. Aspects of their relative stereochemistry were investigated using selective (1)H decoupling, 1D and 2D NOESY, and E-COSY experiments in three different NMR solvents. PMID- 12542363 TI - New dimeric naphthopyrones from Aspergillus niger. AB - Three new dimeric naphthopyrones, asperpyrones A (1), B (2), and C (3), together with two known compounds, fonsecinone A (4) and aurasperone A (5), have been isolated from okara that was fermented with Aspergillus niger JV-33-48. Compounds 1, 4, and 5 showed inhibitory activity on Taq DNA polymerase. PMID- 12542364 TI - A new P-glycoprotein inhibitor from the caper spurge (Euphorbia lathyris). AB - The macrocyclic lathyrane polyester Euphorbia factor L10 (1a) has been obtained from the seeds of the caper spurge (E. lathyris). The interaction of L10 (1a) and its acetyl derivative (1b) with P-glycoprotein, a multidrug transporter overexpressed in cancer cells and responsible for resistance to chemotherapy, was investigated. The results established lathyrane diterpenoids as a novel chemotype for P-glycoprotein inhibitors. PMID- 12542365 TI - Tartrolone C: a novel insecticidal macrodiolide produced by Streptomyces sp. CP1130. AB - A new member of the tartrolone series of macrodiolides, tartrolone C (1), was isolated from a Streptomyces species on the basis of its insecticidal activity. Metacycloprodigiosin (2) and undecylprodigiosin (3) were also isolated on the same basis. The structure of all compounds was established by spectroscopic data (NMR, MS, and UV). PMID- 12542366 TI - Isolation and structure of the cytotoxic cycloheptapeptide phakellistatin 13. AB - A new cyclic heptapeptide phakellistatin 13 (1) had been isolated from the sponge Phakellia fusca Thiele, collected at Yongxing Island of China. Its structure was elucidated as cyclo-(Pro1-Trp-Leu-Thr-Pro2-Gly-Phe) on the basis of MS, UV, IR, and high-field NMR (600 MHz) analysis. The compound was significantly cytotoxic against the human hepatoma BEL-7404 cell line with an ED(50) < 10(-2) microg/mL. PMID- 12542367 TI - A simple synthesis of 7,4'-dihydroxy-6-methoxyisoflavone, glycitein, the third soybean isoflavone. AB - 4-Methoxyresorcinol (3) was synthesized as the precursor for glycitein (6) synthesis by the oxidation of 3-hydroxy-4-methoxybenzaldehyde (1) to the aryl formate with H2O2 and a catalytic amount of SeO2. Glycitein (6) was synthesized by cyclization of 2,4,4'-trihydroxy-5-methoxydeoxybenzoin (5) with N,N dimethylformamide, boron trifluoride diethyl ether, and methanesulfonyl chloride in a microwave oven. PMID- 12542368 TI - New alkaloids from Cephalotaxus fortunei. AB - Four new cephalotaxus alkaloids, cephalotaxine alpha-N-oxide (1), cephalotaxine beta-N-oxide (2), 11-beta-hydroxycephalotaxine beta-N-oxide (3), and isocephalotaxine (4), were isolated, together with several known alkaloids from an EtOAc extract of the fruits of Cephalotaxus fortunei. The structures were determined by spectral analysis including mass spectrometry and 2D NMR. Compounds 1, 2, 3, and 4 displayed cytotoxicity against nasopharynx KB cells with IC50 values of 30, 14, 31, and 15 micro g/mL, respectively. PMID- 12542375 TI - In vivo autofluorescence imaging of early cancers in the human tracheobronchial tree with a spectrally optimized system. AB - The changes in the autofluorescence characteristics of the bronchial tissue is of crucial interest as a cancer diagnostic tool. Evidence exists that this native fluorescence or autofluorescence of bronchial tissues changes when they turn dysplastic and to carcinoma in situ. There is good agreement that the lesions display a decrease of autofluorescence in the green region of the spectrum under illumination with violet-light, and a relative increase in the red region of the spectrum is often reported. Imaging devices rely on this principle to detect early cancerous lesions in the bronchi. Based on a spectroscopic study, an industrial imaging prototype is developed to detect early cancerous lesions in collaboration with the firm Richard Wolf Endoskope GmbH, Germany. A preliminary clinical trial involving 20 patients with this spectrally optimized system shows that the autofluorescence can help to detect most lesions that would otherwise have remained invisible to an experienced endoscopist under white light illumination. A systematic off line analysis of the autofluorescence images pointed out that real-time decisional functions can be defined to reduce the number of false positive results. Using this method, a positive predictive value (PPV) of 75% is reached using autofluorescence only. Moreover, a PPV of 100% is obtained, when combining the white light (WL) mode and the autofluorescence (AF) mode, at the applied conditions. Furthermore, the sensitivity is estimated to be twice higher in the AF mode than in WL mode. PMID- 12542374 TI - Light scattering from cervical cells throughout neoplastic progression: influence of nuclear morphology, DNA content, and chromatin texture. AB - A number of noninvasive fiber optic optical technologies are under development for real-time diagnosis of neoplasia. We investigate how the light scattering properties of cervical cells are affected by changes in nuclear morphology, DNA content, and chromatin texture, which occur during neoplastic progression. We used a Cyto-Savant computer-assisted image analysis system to acquire quantitative nuclear features measurements from 122 Feulgen-thionin-stained histopathologic sections of cervical tissue. A subset of the measured nuclear features was incorporated into a finite-difference time-domain (FDTD) model of cellular light scattering. The magnitude and angular distribution of scattered light was calculated for cervical cells as a function of pathologic grade. The nuclear atypia strongly affected light scattering properties. The increased size and elevated DNA content of nuclei in high-grade lesions caused the most significant changes in scattering intensity. The spatial dimensions of chromatin texture features and the amplitude of refractive index fluctuations within the nucleus impacted both the angular distribution of scattering angles and the total amount of scattered light. Cellular scattering is sensitive to changes in nuclear morphology that accompany neoplastic progression. Understanding the quantitative relationships between nuclear features and scattering properties will aid in the development of noninvasive optical technologies for detection of precancerous conditions. PMID- 12542376 TI - Time-resolved microspectrofluorometry and fluorescence lifetime imaging of photosensitizers using picosecond pulsed diode lasers in laser scanning microscopes. AB - This work describes the time-resolved fluorescence characteristics of two different photosensitizers in single cells, in detail mTHPC and 5-ALA induced PPIX, which are currently clinically used in photodynamic therapy. The fluorescence lifetime of the drugs was determined in the cells from time-gated spectra as well as single photon counting, using a picosecond pulsed diode laser for fluorescence excitation. The diode laser, which emits pulses at 398 nm with 70 ps full width at half maximum duration, was coupled to a confocal laser scanning microscope. For time-resolved spectroscopy a setup consisting of a Czerny Turner spectrometer and a MCP-gated and -intensified CCD camera was used. Time-gated spectra within the cells were acquired by placing the laser beam in "spot scan" mode. In addition, a time-correlated single photon counting module was used to determine the fluorescence lifetime from single spots and to record lifetime images. The fluorescence lifetime of mTHPC decreased from 7.5 to 5.5 ns during incubation from 1 to 6 h. This decrease was probably attributed to enhanced formation of aggregates during incubation. Fluorescence lifetime imaging showed that longer lifetimes were correlated with accumulation in the cytoplasm in the neighborhood of the cell nucleus, whereas shorter lifetimes were found in the outer cytoplasm. For cells that were incubated with 5-ALA, a fluorescence lifetime of 7.4 ns was found for PPIX; a shorter lifetime at 3.6 ns was probably attributed to photoproducts and aggregates of PPIX. In contrast from fluorescence intensity images alone, different fluorescence species could not be distinguished. However, in the lifetime image a structured fluorescence distribution in the cytoplasm was correlated with the longer lifetime and probably coincides with mitochondria. In conclusion, picosecond diode lasers coupled to a laser scanning microscope equipped with appropriate detection units allows time-resolved spectroscopy and lifetime imaging with high spatial resolution and provides numerous possibilities in cellular and pharmaceutical research. PMID- 12542377 TI - Feasibility study using surface-enhanced Raman spectroscopy for the quantitative detection of excitatory amino acids. AB - The release of excitatory amino acids (EAAs) from injured neurons has been associated with secondary injury following head trauma. The development of a rapid and sensitive method for the quantification of EAAs may provide a means for clinical management of patients affected by head trauma. We explore the potential application of surface-enhanced Raman spectroscopy (SERS) for rapid quantification of the concentration of EAAs in aqueous silver colloids. The EAAs glutamate (Glu) and aspartate (Asp) are released following head injury and have been observed to exhibit SERS spectra that should enable them to be distinguished in a complex aqueous media. Of the two EAAs, the concentration of Glu has been shown to be more indicative of injury to the central nervous system. Using 30-s scans and a 50-mW argon laser, aqueous Glu is quantifiable from 0.4 to 5 micromol/L and is spectrally distinguishable from Asp. In addition, initial in vivo microdialysis experiments suggest that this SERS system is capable of measuring chemical changes following head trauma in the rat brain. Compared with current high-performance liquid chromatography (HPLC) techniques for amino acid detection, the short scanning and processing time associated with the SERS approach enables measurement on a near-real-time basis, providing clinical information in anticipation of pharmaceutical intervention. PMID- 12542378 TI - Spatiotemporal anisotropic diffusion filtering to improve signal-to-noise ratios and object restoration in fluorescence microscopic image sequences. AB - We present an approach for significantly improving the quantitative analysis of motion in noisy fluorescence microscopic image sequences. The new partial differential equation based method is a general extension of a 2-D nonlinear anisotropic diffusion filtering scheme to a specially adapted 3-D nonlinear anisotropic diffusion filtering scheme, with two spatial image dimensions and the time t in the image sequence as the third dimension. Motion in image sequences is considered as oriented, line-like structures in the spatiotemporal x,y,t domain, which are determined by the structure tensor method. Image enhancement is achieved by a structure adopted smoothing kernel in three dimensions, thereby using the full 3-D information inherent in spatiotemporal image sequences. As an example for low signal-to-noise ratio (SNR) microscopic image sequences we have applied this method to noisy in vitro motility assay data, where fluorescently labeled actin filaments move over a surface of immobilized myosin. With the 3-D anisotropic diffusion filtering the SNR is significantly improved (by a factor of 3.8) and closed object structures are reliably restored, which were originally degraded by noise. Generally, this approach is very valuable for all applications where motion has to be measured quantitatively in low light level fluorescence microscopic image sequences of cellular, subcellular, and molecular processes. PMID- 12542379 TI - Kinetics of picosecond laser pulse induced charge separation and proton transfer in bacteriorhodopsin. AB - Bacteriorhodopsin (BR) films oriented by an electrophoretic method are deposited on a transparent conductive ITO glass. A counterelectrode of copper and gelose gel is used to compose a sandwich-type photodetector with the structure of ITO/BR film/gelose gel/Cu. A single 30-ps laser pulse and a mode-locked pulse train are respectively used to excite the BR photodetector. The ultrafast falling edge and the bipolar response signal are measured by the digital oscilloscope under seven different time ranges. Marquardt nonlinear least squares fitting is used to fit all the experimental data and a good fitting equation is found to describe the kinetic process of the photoelectric signal. Data fitting resolves six exponential components that can be assigned to a seven-step BR photocycle model: BR-->K-->KL-->L-->M-->N-->O-->BR. Comparing tests of the BR photodetector with a 100-ps Si PIN photodiode demonstrates that this type of BR photodetector has at least 100-ps response time and can also serve as a fast photoelectric switch. PMID- 12542380 TI - Interplay of tumor vascular oxygenation and tumor pO2 observed using near infrared spectroscopy, an oxygen needle electrode, and 19F MR pO2 mapping. AB - This study investigates the correlation of tumor blood oxygenation and tumor pO(2) with respect to carbogen inhalation. After having refined and validated the algorithms for calculating hemoglobin concentrations, we used near-infrared spectroscopy (NIRS) to measure changes of oxygenated hemoglobin concentration (delta[HbO(2)]) and used an oxygen needle electrode and (19)F MRI for pO(2) measurements in tumors. The measurements were taken from Dunning prostate R3327 tumors implanted in rats, while the anesthetized rats breathed air or carbogen. The NIRS results from tumor measurements showed significant changes in tumor vascular oxygenation in response to carbogen inhalation, while the pO(2) electrode results showed an apparent heterogeneity for tumor pO(2) response to carbogen inhalation, which was also confirmed by (19)F MR pO(2) mapping. Furthermore, we developed algorithms to estimate hemoglobin oxygen saturation, sO(2), during gas intervention based on the measured values of delta[HbO(2)] and pO(2). The algorithms have been validated through a tissue-simulating phantom and used to estimate the values of sO(2) in the animal tumor measurement based on the NIRS and global mean pO(2) values. This study demonstrates that the NIRS technology can provide an efficient, real-time, noninvasive approach to monitoring tumor physiology and is complementary to other techniques, while it also demonstrates the need for an NIR imaging technique to study spatial heterogeneity of tumor vasculature under therapeutic interventions. PMID- 12542381 TI - Blood pO2 and blood flow at the optic disc. AB - A fundus camera-based phosphorometer to noninvasively and quasicontinuously measure the blood partial pressure of oxygen (pO(2,blood)) in the microvasculature of the pig optic nerve using the principle of the phosphorescence quenching by O(2) is described. A porphyrin dye is injected into the venous circulation and the decay of its phosphorescence emission is detected locally in the eye, after excitation with a flash of light. Combined with blood flow measurements by means of a laser Doppler flowmeter mounted on the phosphorometer, we demonstrate the capability of the instrument to determine the time course of optic nerve blood flow and pO(2,blood) in response to various physiological stimuli, such as hyperoxia and hypercapnia. This instrument appears to be a useful tool for the investigation of the oxygenation of the optic nerve. PMID- 12542382 TI - Light-scattering instrument to detect thromboemboli in blood. AB - The characteristics and capabilities of a light-scattering microemboli detector (LSMD) are delineated by detailing its state-of-the-art configuration, by discussing the theoretical and empirical aspects of instrument calibration, and by summarizing various experimental studies that have benefited from this instrument. In the past, thromboembolism, which often results when blood contacts medical devices, has eluded scientific scrutiny due to the absence of instruments that could detect and quantify thromboemboli in circulating blood. More recently, the ability of the LSMD to provide continuous, noninvasive detection of thromboemboli in whole blood (meaning that the LSMD probe does not contact the blood) was exploited in various in vitro and ex vivo models to explore thromboembolic phenomena. Through this work, the LSMD evolved as a sensitive and an economical research tool for the study of thromboembolic phenomena. PMID- 12542383 TI - Measurement of skin stretch via light reflection. AB - A noninvasive technique for measuring the stretch of skin is described. The technique utilizes changes in the reflectivity of polarized light intensity as a monitor of skin stretch. Measurements of in vitro pigskin and in vivo human skin show that the reflectivity of polarized light intensity increases linearly with stretch. The changes in diffusive reflectivity properties of skin result from the alterations that take place in the roughness across the thickness of the skin layers due to stretch. Conceptually, as the roughness of a layer decreases with stretch, a smoother reflecting media is produced, resulting in a proportional increase in the specular reflection. Results can be easily extended to a real time stretch analysis of large tissue areas that would be applicable for mapping the stretch of skin. PMID- 12542384 TI - Assessment of an in situ temporal calibration method for time-resolved optical tomography. AB - A 32-channel time-resolved optical imaging device is developed at University College London to produce functional images of the neonatal brain and the female breast. Reconstruction of images using time-resolved measurements of transmitted light requires careful calibration of the temporal characteristics of the measurement system. Since they can often vary over a period of time, it is desirable to evaluate these characteristics immediately after, or prior to, the acquisition of image data. A calibration technique is investigated that is based on the measurement of light back-reflected from the surface of the object being imaged. This is facilitated by coupling each detector channel with an individual source fiber. A Monte Carlo model is employed to investigate the influence of the optical properties of the object on the back-reflected signal. The results of simulations indicate that their influence may be small enough to be ignored in some cases, or could be largely accounted for by a small adjustment to the calibrated data. The effectiveness of the method is briefly demonstrated by imaging a solid object with tissue-equivalent optical properties. PMID- 12542385 TI - Treatment of nevus using medical tattooing. AB - Medical tattooing is used to color skin with a pigment loss. Currently, however, a trial-and-error scheme is employed to obtain the desired color appearance of tattooed skin because prediction of the color appearance is dependent on the experiences of medical doctors. We propose a method for predicting the color appearance of tattooed skin. Two trial dyes are first injected in the area of pigment loss, and the color appearance of a third dye to be injected can be predicted using measured spectrocolorimeter data and mathematical formula. The spectrocolorimeter measures the color appearances of the skin before and after tattooing using the first two dyes, and the mathematical formula calculates the color appearance of the tattooed skin using any third dye. In the derivation of the mathematical formula, light propagation in the skin has been modeled using the modified Lambert-Beer law considering the strong scattering of light by biological tissues. The proposed method was successfully validated by a preliminary tattooing of the skin to an area with pigment loss. Predicting the color appearance of tattooed skin significantly reduces the number of trial-and error attempts required in the current methods. Medical tattooing using this method can also be applied to treat various skin color abnormalities such as leukoderma, intradermal nevi, and reconstructed nipples. PMID- 12542386 TI - Near-infrared imaging in the small animal brain: optimization of fiber positions. AB - We investigate fiber placement issues associated with a hybrid magnetic resonance imaging (MRI) near-infrared (NIR) imaging technique for small animal brain studies. Location of the optical fibers on the cranium is examined, with an emphasis on maximizing the recovered resolution and contrast in the region of interest, which in this case is the murine brain. In a series of simulation studies, singular value decomposition of the Jacobian is used in order to determine the measurement sites that provide the most information about the region of interest. The modeling results indicate that data collected using fibers arranged on one side of the head near the brain contain as much information about optical changes within the brain as those positioned equally spaced around the entire periphery of the head. Practical space limitation considerations favor the one-sided fiber array geometry in the case where the NIR acquisition is expected to occur simultaneously with MRI. PMID- 12542387 TI - Near-infrared fluorescence contrast-enhanced imaging with intensified charge coupled device homodyne detection: measurement precision and accuracy. AB - Fluorescence frequency-domain photon migration (FDPM) through tissue refers to the propagation of intensity-modulated fluorescent light that originates from tissue-laden fluorophores following illumination with an intensity-modulated excitation light source. FDPM measurements of modulation amplitude and phase are ultimately employed in an inversion algorithm for tomographic reconstruction of interior optical and fluorescent property maps that delineate disease enhanced with fluorescent contrast agent. Because the inverse problem is underdetermined, measurement precision and accuracy crucially impact its solution. Reported here are the precision and accuracy of FDPM measurements acquired using an intensified CCD homodyne detection system. By introducing 32 phase delays between the oscillators used to modulate the intensifier gain and light source intensity at 100 MHz, mean precision is maximized at +/-0.46% and +/-0.26 deg for measurements of modulation amplitude and phase, respectively. Measurement precision improves when the number of phase delays increases. Measurements of fluorescence modulation amplitude and phase, acquired from the surface of a tissue phantom at distances ranging between 0.71 and 3.6 cm from an incident excitation point source, exhibit a mean accuracy of 17% and 1.9 deg, respectively. Measurement accuracy deteriorates with increasing distance from the point source, but for distances up to 1.0 cm from the point source, measurements of fluorescence modulation amplitude and phase exhibit a mean accuracy of 5.4% and 0.30 deg, respectively. PMID- 12542388 TI - Fiber optic probes for biomedical optical spectroscopy. AB - Fiber optic probes are a key element for biomedical spectroscopic sensing. We review the use of fiber optic probes for optical spectroscopy, focusing on applications in turbid media, such as tissue. The design of probes for reflectance, polarized reflectance, fluorescence, and Raman spectroscopy is illustrated. We cover universal design principles as well as technologies for beam deflecting and reshaping. PMID- 12542389 TI - Liquid light guides versus fiber light guides in clinical near-infrared spectroscopy. AB - New commercial liquid light guides have an advantage over fiberoptic bundles regarding breakage during clinical handling. We investigate the quality of clinical data collection using liquid versus fiber bundles as receivers. A four wavelength NIRO-500 near-IR spectrophotometer is used with single-terminal fiber bundles, multiterminal fiber bundles, or a single-terminal liquid light guide as receivers. Repeated 3-min trials are done using a stable phantom, an unstable phantom, and the human forearm. A least-squares linear best-fit line and its root mean square error (RMSE), a measure of signal noise, are derived for each wavelength of each trial. The mean and standard deviations for the RMSEs of the single-terminal fiber optic receiving cable are derived for comparison standards. The liquid light guides have 51 to 174% greater signal noise with RMSEs 2 to 12 standard deviations above the mean of the single-terminal fiber bundle. The multiterminal fiber bundles have 49% less to 32% greater signal noise and had RMSEs within 1 to 4 standard deviations above the mean of the single-terminal fiber bundle. These comparisons suggest fiber optic bundles are preferable for clinical near-IR spectroscopy (NIRS) applications requiring low signal noise. PMID- 12542390 TI - Self-mixing in a diode laser as a method for cardiovascular diagnostics. AB - Our aim is the development of a simple optical method for pulse wave profile, pulse wave delay time, and blood flow measurement. The method is based on recording the Doppler frequency shift related to a moving target--blood vessel walls or small particles. The Doppler signal is detected using the self-mixing that occurs in the diode laser cavity when radiation scatters back from the moving target into the laser and interferes with the field inside. Two different ways can be simultaneously used for the self-mixing signal extraction: a photodiode accommodated in the rear facet of the diode laser package and a resistor from the laser pump current. An experimental device with a pigtail laser diode is developed that is able to detect the pulsation of major arteries with potentially useful information, including the pulse wave profile and the pulse wave delay time. The pulse wave delay time in different regions of the human body is measured relative to the electrocardiogram (ECG) signal. Also the flow velocity of a liquid suspension containing particles the size of erythrocytes (equivalent to blood flow) is measured. Registered signals are stored after digitalization and preprocessed using LabView for a Windows environment. The described device has the application of the self-mixing method and highlights significant advantages of simplicity, compactness, and robustness as well as the self-aligning and self-detecting abilities of such method, compared with the use of conventional interferometric method. PMID- 12542391 TI - [Infection prevention and safe practice (referring to CDC guideline)]. AB - Recent nosocomial infections, such as HCV and tuberculosis, have stimulated interest in infection control practices in health care setting. Therefore, Infection Control Doctors (ICDs) have already started to make best strategies for infection prevention and safe practice. However, many obstacles lie ahead of them, with the cost as the most important. It has been believed that it takes a lot of money for the improvement of infection control. But, this aspect has been changed completely by introduction of EBM because EBM can point out and quite unnecessary practices, resulting in saving money. CDC has been published many guidelines which is evidence based. Therefore, it might be best strategy to introduce CDC guideline into the infection control in Japanese hospitals. PMID- 12542392 TI - [Simultaneous detection of hepatitis B virus surface antigen (HBsAg) and hepatitis C virus (HCV) related antibodies by an immunochromatographic test method using synthesized oligonucleotide-bound protein probes, OligoFast]. AB - An immunochromatographic test using synthesized oligonucleotide-bound protein probes, OligoFast (Nissui Pharmaceutical Co., Ltd., Tokyo) was developed and evaluated for simultaneous detection of hepatitis B surface antigen (HBsAg) and antibodies related with hepatitis C virus (HCV). The color development of colloidal gold was visually read and easily interpretable for the respective antigen and antibody, positive or negative. When the performance panels of Boston Biomedica Inc. (BBI) for HBsAg and HCV-related antibodies were assayed, the results indicated; first, the most positive specimens with 1.2 IU/ml of HBsAg were correctly determined as positive, and secondly, all the positive specimens for HCV-related antibodies confirmed with Ortho RIBA 3.0 were consistently determined as positive and additional two undetermined specimens were interpreted as positive. However, when the seroconversion panel of BBI for hepatitis B virus (HBV) infection, the seroconversion was delayed 20 to 30 days when compared to HBV DNA detection. When the clinical serum specimens were tested in comparison with the automated AxSYM (Abbott Laboratories, Abbott Park, IL, U.S.A.), both sensitivity and specificity were estimated to be 100% for HBsAg, and 91.3% and 99.0% for HCV-related antibodies, respectively. With these results, we can conclude that this newly developed immunochromatographic test will be applicable to simultaneous detection of HBsAg and HCV-related antibodies in a single device, and will be expected to be widely applied in a clinical setting. PMID- 12542393 TI - [Evaluation of Norwalk virus detection kit by enzyme immunoassay]. AB - We briefly examined detection kit using the EIA method for Norwalk virus, and compared the results of the tests using the EIA method with those using RT-PCR method. In reproducibility, an amount of variation was observed in data obtained from positive controls and in lower values. The sensitivity obtained from the EIA method was about 300 times lower than that obtained from the RT-PCR method. Results accordance ratio between EIA method and RT-PCR method was 70%. This results discrepancy was presumably caused by a difference of sensitivity and specificity between these two methods. In conclusion, this detection kit using the EIA method is easily manually operated so that this kit can be considered as an effective and simple detection tool for Norwalk virus. PMID- 12542394 TI - [The cyclopedic detection of pathogens from digestive and respiratory system using polymerase chain reaction (PCR) and DNA microarray]. PMID- 12542395 TI - [Practical evaluation of rapid diagnostic tests for mycobacteriosis and virus infection]. AB - Because cultures of mycobacteria and viruses take a great number of days, the results are delayed in these clinical laboratory tests. For this reason, rapid diagnostic tests without these cultures are required. Among the laboratory tests of mycobacteria, acid-fast stain as a rapid diagnostic test is the first to give significant information to laboratory staffs and physicians. There are mainly two procedures, directly smearing clinical specimens and smearing concentrated specimens treated with NALC-NaOH on slide glass. The former is performed for specimens from emergency patients. When a patient is suspected of tuberculosis, a DNA-amplification test such as COBAS AMPLICOR TEST (PCR test) is performed for directly detecting Mycobacterium tuberculosis in clinical specimens, regardless of whether the specimens show positive or negative by acid-fast stain. Sixty-one of 1,023 specimens tested in Showa University Fujigaoka Hospital in February 2000 May 2002 were found positive by both PCR and culture tests. Fourteen were found negative by PCR test and positive by culture test. Five were found positive by PCR test and negative by culture test. Alternatively, MTB-64 (Capilia TB) is a kit for rapidly identifying M. tuberculosis using colonies. The time necessary for completion of the test was approximately 15 minutes per sample. In our study, the identification test showed weak-positive for 2 strains of M. marinum (ATCC927 and one clinical isolate), compared with positive for 28 clinical isolates and strain H37Ra of M. tuberculosis. Recently, rapid diagnostic kits for detecting various pathogenic viruses have been developed. The kits for detecting influenza virus are in especially great demand with the spread of newly-developed antiviral agents. There was an epidemic of the type A virus in the winters 2001-2002. 101 (32%) of 316 clinical specimens tested in the hospital were found positive with InfluA. B-Quick [SEIKEN] (Denka) or Directigen Flu A (Becton Dickinson). These kits showed high utility. Also, Abbot TESTPACK RSV (Dainabot) for detecting RS virus and ROTALEX DRY (Orion Diagnostica) for detecting rotavirus showed high utility. However, these rapid diagnostic kits have weak points: high costs, false positives and false negatives. Thus, more improvements are required. PMID- 12542396 TI - Deglycosylation, processing and crystallization of human testis angiotensin converting enzyme. AB - Angiotensin I-converting enzyme (ACE) is a highly glycosylated type I integral membrane protein. A series of underglycosylated testicular ACE (tACE) glycoforms, lacking between one and five N-linked glycosylation sites, were used to assess the role of glycosylation in tACE processing, crystallization and enzyme activity. Whereas underglycosylated glycoforms showed differences in expression and processing, their kinetic parameters were similar to that of native tACE. N glycosylation of Asn-72 or Asn-109 was necessary and sufficient for the production of enzymically active tACE but glycosylation of Asn-90 alone resulted in rapid intracellular degradation. All mutants showed similar levels of phorbol ester stimulation and were solubilized at the same juxtamembrane cleavage site as the native enzyme. Two mutants, tACEDelta36-g1234 and -g13, were successfully crystallized, diffracting to 2.8 and 3.0 A resolution respectively. Furthermore, a truncated, soluble tACE (tACEDelta36NJ), expressed in the presence of the glucosidase-I inhibitor N -butyldeoxynojirimycin, retained the activity of the native enzyme and yielded crystals belonging to the orthorhombic P2(1)2(1)2(1) space group (cell dimensions, a=56.47 A, b=84.90 A, c=133.99 A, alpha=90 degrees, beta=90 degrees and gamma=90 degrees ). These crystals diffracted to 2.0 A resolution. Thus underglycosylated human tACE mutants, lacking O-linked oligosaccharides and most N-linked oligosaccharides or with only simple N-linked oligosaccharides attached throughout the molecule, are suitable for X-ray diffraction studies. PMID- 12542397 TI - Generation of hepatocytes expressing functional cytochromes P450 from a pancreatic progenitor cell line in vitro. AB - The proliferating AR42J-B13 pancreatic cell line is known to respond to glucocorticoid treatment by producing foci of cells that express the liver specific albumin gene. We demonstrate that this cell line also expresses liver specific or liver-enriched functional cytochrome P450 proteins when stimulated to trans-differentiate into hepatocytes by glucocorticoid. These data suggest that this cell line has an unusual ability to trans-differentiate into functional hepatocytes and that it could be possible to generate a limitless supply of functional hepatocyte-like cells in vitro. PMID- 12542398 TI - A peptide mimetic of human interferon (IFN)-beta. AB - Type I interferons (IFNs) are cytokines that are used clinically as antiviral and antitumour agents. The interaction of IFNs with their heterodimeric type I IFN receptor comprised of IFNAR1 and IFNAR2 is a first step to inducing biological actions. Here, we describe the successful mimicry of IFN-beta by a peptide isolated by phage-display screening using a neutralizing anti-IFN-beta monoclonal antibody. The 15-mer peptide, designated SYR6, was shown to compete with IFN-beta for binding to type I IFN receptor in a concentration-dependent manner, and was shown to elicit antiviral activity on cultured cells. This antiviral activity was not eliminated in the presence of neutralizing monoclonal antibodies to IFN alpha, -beta and -gamma, and a low concentration of soluble type I IFN receptor, suggesting that it was not due to IFN contamination or the induction of endogenous IFNs by SYR6. This peptide might be a potent agonist to provide a mechanism of activating heterodimeric cytokine receptors. PMID- 12542399 TI - Chipping away at breast cancer: insights from microarray studies of human and mouse mammary cancer. AB - Breast cancer is the most prevalent tumor in American woman. Multiple factors, including age, diet, genetics, environment, geographic location, parity, as well as race, influence the development of this heterogeneous disease. As the process of oncogenesis involves the disruption of diverse cellular pathways including cell cycle, growth, survival, and apoptosis, the high throughput technique of microarray analyses provides a powerful insight into multiple cellular processes. These techniques have identified particular expression patterns that can classify tumors into new groups and aid in the prediction of the natural history of the disease and the therapeutic response. This wealth of information may also form the basis for the development of new types of targeted therapies. Studies to identify the earliest molecular events in oncogenesis and progressive changes in the human disease have been difficult to perform within the same patient. The use of transgenic mouse mammary cancer models provides an opportunity to decipher molecular changes that occur at progressive stages of tumor development. This paper reviews microarray technology, and the insights gained from published breast cancer microarray analyses, and considers the contribution of microarray studies in identifying mouse cancer models that may be appropriate for answering particular experimental questions. PMID- 12542400 TI - Uncontrolled insulin secretion from a childhood pancreatic beta-cell adenoma is not due to the functional loss of ATP-sensitive potassium channels. AB - We report the case of an 8-year-old child who presented with severe hyperinsulinaemic hypoglycaemia due to a pancreatic islet cell adenoma. In vivo, there was no beneficial response to the hyperglycaemia-inducing agent diazoxide and as a consequence the child underwent a subtotal pancreatectomy. In vitro studies of adenomatous beta-cells revealed no operational defects in ATP sensitive potassium channel activity and appropriate responses to diazoxide. In comparison with patients with focal adenomatous hyperplasia, genetic analysis of the isolated adenoma showed no loss of heterozygosity for chromosome 11p15 and expression of the cyclin-dependent kinase inhibitor p57(kip2). This case illustrates that the excess insulin secretion from an infantile adenoma has an aetiology different from that observed in hyperinsulinism in infancy. PMID- 12542401 TI - Management of papillary and follicular (differentiated) thyroid cancer: new paradigms using recombinant human thyrotropin. AB - The incidence of differentiated thyroid cancer (DTC) has increased in many places around the world over the past three decades, yet this has been associated with a significant decrease in DTC mortality rates in some countries. While the best 10 year DTC survival rates are about 90%, long-term relapse rates remain high, in the order of 20-40%, depending upon the patient's age and tumor stage at the time of initial treatment. About 80% of patients appear to be rendered disease-free by initial treatment, but the others have persistent tumor, sometimes found decades later. Optimal treatment for tumors that are likely to relapse or cause death is total thyroidectomy and ablation by iodine-131 ((131)I), followed by long-term levothyroxine suppression of thyrotropin (TSH). On the basis of regression modeling of 1510 patients without distant metastases at the time of initial treatment and including surgical and (131)I treatment, the likelihood of death from DTC is increased by several factors, including age >45 years, tumor size >1.0 cm, local tumor invasion or regional lymph-node metastases, follicular histology, and delay of treatment >12 months. Cancer mortality is favorably and independently affected by female sex, total or near-total thyroidectomy, (131)I treatment and levothyroxine suppression of TSH. Treatments with (131)I to ablate thyroid remnants and residual disease are independent prognostic variables favorably influencing distant tumor relapse and cancer death rates. Delay in treatment of persistent disease has a profound impact on outcome. Optimal long term follow-up using serum thyroglobulin (Tg) measurements and diagnostic whole body scans (DxWBS) require high concentrations of TSH, which until recently were possible to achieve only by withdrawing levothyroxine treatment, producing symptomatic hypothyroidism. New paradigms, however, provide alternative pathways to prepare patients for (131)I treatment and to optimize follow-up. Patients with undetectable or low Tg concentrations and persistent occult disease can now be identified within the first year after initial treatment by recombinant human (rh)TSH-stimulated serum Tg concentrations greater than 2 microg/l, without performing DxWBS. These new follow-up paradigms promptly identify patients with lung metastases that are not evident on routine imaging, but which respond to (131)I treatment. In addition, rhTSH can be given to prepare patients for (131)I remnant ablation or (131)I treatment for metastases, especially those who are unable to withstand hypothyroidism because of concurrent illness or advanced age, or whose hypothyroid TSH fails to increase. PMID- 12542402 TI - Molecular targets in breast cancer: current status and future directions. AB - Our increasing understanding of the pathophysiologic mechanisms of breast carcinogenesis has generated detailed information about the potential roles of specific biomolecular markers in this process. Furthermore, in the last few years the process of targeted drug design has become faster and more sophisticated, providing a variety of agents targeted at these molecules. In this review, we describe the most widely recognized molecular targets in breast cancer. PMID- 12542403 TI - Fulvestrant ('Faslodex')--a new treatment option for patients progressing on prior endocrine therapy. AB - Since its introduction more than 30 years ago, tamoxifen has been the most widely used endocrine therapy for the treatment of women with advanced breast cancer. More recently, a number of alternative endocrine treatments have been developed, including several selective estrogen receptor modulators (SERMs), aromatase inhibitors (AIs) and, most recently, fulvestrant ('Faslodex'). Fulvestrant is an estrogen receptor (ER) antagonist, which, unlike the SERMs, has no known agonist (estrogenic) effect and downregulates the ER protein. Tamoxifen is effective and well tolerated, although the non-steroidal AIs, anastrozole and letrozole, are more effective treatments for advanced disease than tamoxifen. Fulvestrant has recently gained US Food and Drug Administration approval for the treatment of hormone receptor-positive metastatic breast cancer in postmenopausal women with disease progression following antiestrogen therapy. In two global phase III clinical trials fulvestrant was at least as effective and as equally well tolerated as anastrozole for the treatment of postmenopausal women with advanced and metastatic breast cancer. In a retrospective analysis of the combined data from these trials, mean duration of response was significantly greater for fulvestrant compared with anastrozole. These new hormonal treatments expand the choice of endocrine therapy for women with advanced breast cancer and offer new options for sequencing and combining treatments. PMID- 12542404 TI - Ontologies for molecular biology and bioinformatics. AB - About five years ago, ontology was almost unknown in bioinformatics, even more so in molecular biology. Nowadays, many bioinformatics articles mention it in connection with text mining, data integration or as a metaphysical cure for problems in standardisation of nomenclature and other applications. This article attempts to give an account of what concept ontologies in the domain of biology and bioinformatics are; what they are not; how they can be constructed; how they can be used; and some fallacies and pitfalls creators and users should be aware of. PMID- 12542405 TI - AGenDA: gene prediction by comparative sequence analysis. AB - Comparative sequence analysis is a powerful approach to identify functional elements in genomic sequences. Herein, we describe AGenDA (Alignment-based GENe Detection Algorithm), a novel method for gene prediction that is based on long range alignment of syntenic regions in eukaryotic genome sequences. Local sequence homologies identified by the DIALIGN program are searched for conserved splice signals to define potential protein-coding exons; these candidate exons are then used to assemble complete gene structures. The performance of our method was tested on a set of 105 human-mouse sequence pairs. These test runs showed that sensitivity and specificity of AGenDA are comparable with the best gene- prediction program that is currently available. However, since our method is based on a completely different type of input information, it can detect genes that are not detectable by standard methods and vice versa. Thus, our approach seems to be a useful addition to existing gene-prediction programs. AVAILABILITY: DIALIGN is available through the Bielefeld Bioinformatics Server (BiBiServ) at http://bibiserv.techfak.uni-bielefeld.de/dialign/ The gene-prediction program AGenDA described in this paper will be available through the BiBiServ or MIPS web server at http://mips.gsf.de. PMID- 12542406 TI - A system architecture for genomic data analysis. AB - MOTIVATION: Most of diseases are caused by a set of gene defects, which occur in a complex association. The association scheme of expressed genes can be modelled by genetic networks. Genetic networks are efficiently facilities to understand the dynamic of pathogenic processes by modelling molecular reality of cell conditions. In this sense a genetic network consists of first, a set of genes of specified cells, tissues or species and second, causal relations between these genes determining the functional condition of the biological system, i. e. under disease. A relation between two genes will exist if they both are directly or indirectly associated with disease [8]. Our goal is to characterize diseases (especially autoimmune diseases like chronic pancreatitis CP, multiple sclerosis MS, rheumatoid arthritis RA) by genetic networks generated by a computer system. We want to introduce this practice as a bioinformatic approach for finding targets. PMID- 12542407 TI - Building a genome database using an object-oriented approach. AB - GOBASE is a relational database that integrates data associated with mitochondria and chloroplasts. The most important data in GOBASE, i. e., molecular sequences and taxonomic information, are obtained from the public sequence data repository at the National Center for Biotechnology Information (NCBI), and are validated by our experts. Maintaining a curated genomic database comes with a towering labor cost, due to the shear volume of available genomic sequences and the plethora of annotation errors and omissions in records retrieved from public repositories. Here we describe our approach to increase automation of the database population process, thereby reducing manual intervention. As a first step, we used Unified Modeling Language (UML) to construct a list of potential errors. Each case was evaluated independently, and an expert solution was devised, and represented as a diagram. Subsequently, the UML diagrams were used as templates for writing object oriented automation programs in the Java programming language. PMID- 12542408 TI - The semantic metadatabase (SEMEDA): ontology based integration of federated molecular biological data sources. AB - A system for "intelligent" semantic integration and querying of federated databases is being implemented by using three main components: A component which enables SQL access to integrated databases by database federation (MARGBench), an ontology based semantic metadatabase (SEMEDA) and an ontology based query interface (SEMEDA-query). In this publication we explain and demonstrate the principles, architecture and the use of SEMEDA. Since SEMEDA is implemented as 3 tiered web application database providers can enter all relevant semantic and technical information about their databases by themselves via a web browser. SEMEDA' s collaborative ontology editing feature is not restricted to database integration, and might also be useful for ongoing ontology developments, such as the "Gene Ontology" [2]. SEMEDA can be found at http://www-bm.cs.uni magdeburg.de/semeda/. We explain how this ontologically structured information can be used for semantic database integration. In addition, requirements to ontologies for molecular biological database integration are discussed and relevant existing ontologies are evaluated. We further discuss how ontologies and structured knowledge sources can be used in SEMEDA and whether they can be merged supplemented or updated to meet the requirements for semantic database integration. PMID- 12542409 TI - Construction of stochastic context trees for genetic texts. AB - A method has been developed for constructing a tree source model for genetic text generation. Model visualisation in the form of suffix (context) trees provides a new way of context analysis of symbol sequences. Estimation of the stochastic complexity of the data in the frame of the model serves as a criterion for the model's ascertainment. The model and complexity values are used for analysis of genetic texts. The software realisation of this algorithm enables to reveal statistical properties of genetic sequences based on an information measure. The program developed is available via Internet at http://wwwmgs.bionet.nsc.ru/mgs/programs/complexity/. PMID- 12542410 TI - Finding and decrypting of promoters contributes to the elucidation of gene function. AB - The combination of full-scale genomic sequencing with high throughput expression analysis provides a new and largely unexploited basis for in silico functional genomics. Recent break through developments in locating and analyzing promoters now allow extending functional genomics in silico far beyond identification of protein sequences into the complex regulatory structures and mechanisms of the genome. However, only first examples of this new type of approach are emerging at present and intensive further developments of bioinformatics tools will be required before such analysis can become large-scale routine in genomic sequence analysis. Nevertheless, the door to a new dimension of functional analysis of the genomic sequence is open. Finally, only the tight integration of the enormous amount of knowledge gained from proteins sequence analysis with the complementary information about gene regulation will afford us with a more complete picture of the networks than constitute life. PMID- 12542411 TI - Computer system "gene discovery" for promoter structure analysis. AB - This paper presents implementation of Data Mining and Knowledge Discovery techniques for searching for regularities in tables of context features of DNA sequences involved in regulation of transcription. The goal is to discover regularities that relate nucleotide sequences to the functional classes of these sequences. The search patterns for regularities have been constructed in the first-order logic augmented by probabilistic estimates. To this aim. the PC software system "Gene Discovery" has been designed. This system accepts molecular genetical data retrieved from a database by using SQL queries. Nucleotide sequences of promoters of several functional systems were extracted from the TRRD database (http://wwwmgs.bionet.nsc.ru/mgs/gnw/trrd/) and analysed. The data include nucleotide sequences of erythroid-specific gene promoters, endocrine system gene promoters, promoter regions of the genes controlling cell cycle, promoter of genes regulating lipid metabolism, and muscle-specific gene promoters. Several regularities that relate the nucleotide sequences in the regulatory DNA and their location relative to the transcription start with each functional class have been found. PMID- 12542412 TI - Mining putative regulatory elements in promoter regions of Saccharomyces cerevisiae. AB - The availability of genome-wide gene expression data provides a unique set of genes from which we can decipher the mechanisms underlying the common transcriptional response. Transcription factors, which can bind to specific DNA sites, cooperatively regulate the transcription of genes. This study attempts to mine putative binding sites to investigate how combinations of the sites predicted from known sites and over-represented repetitive elements are distributed in the promoter regions of groups of functionally related genes. The over-represented repetitive elements appearing in the associations are possible transcription factor binding sites. The deduced association rules would facilitate to predict putative regulatory elements and to identify genes which are potentially co-regulated by the putative regulatory elements. Our proposed approach is applied to Saccharomyces cerevisiae and the promoter regions of yeast ORFs. PMID- 12542413 TI - Protein similarity search under mRNA structural constraints: application to targeted selenocysteine insertion. AB - Selenocysteine is the 21th amino acid, which occurs in all kingdoms of life. Selenocysteine is encoded by the STOP-codon UGA. For its insertion, it requires a specific mRNA sequence downstream the UGA-codon that forms a hairpin like structure (called Sec insertion sequence (SECIS)). We consider the computational problem of generating new amino acid sequences containing selenocysteine. This requires to find an mRNA sequence that is similar to the SECIS-consensus, is able to form the secondary structure required for selenocysteine insertion, and whose translation is maximally similar to the original amino acid sequence. We show that the problem can be solved in linear time when considering the hairpin-like SECIS-structure (and, more generally, when considering a structure that does not contain pseudoknots). PMID- 12542414 TI - An overview on predicting the subcellular location of a protein. AB - The present paper overviews the issue on predicting the subcellular location of a protein. Five measures of extracting information from the global sequence based on the Bayes discriminant algorithm are reviewed. 1) The auto-correlation functions of amino acid indices along the sequence; 2) The quasi-sequence-order approach; 3) the pseudo-amino acid composition; 4) the unified attribute vector in Hilbert space, 5) Zp parameters extracted from the Zp curve. The actual performance of the predictive accuracy is closely related to the degree of similarity between the training and testing sets or to the average degree of pairwise similarity in dataset in a cross-validated study. Many scholars considered that the current higher predictive accuracy still cannot ensure that some available algorithms are effective in practice prediction for the higher pairwise sequence identity of the datasets, but some of them declared that construction of the dataset used for developing software should base on the reality determined by the Mother Nature that some subcellular locations really contain only a minor number of proteins of which some even have a high percentage of sequence similarity. Owing to the complexity of the problem itself, some very sophisticated and special programs are needed for both constructing dataset and improving the prediction. Anyhow finding the target information in mature protein sequence and properly cooperating it with sorting signals in prediction may further improve the overall predictive accuracy and make the prediction into practice. PMID- 12542415 TI - Molecular dynamics simulations on the free and complexed N-terminal SH2 domain of SHP-2. AB - Signal transduction events are often mediated by small protein domains such as SH2 (Src homology 2) domains that recognize phosphotyrosines (pY) and flanking sequences. In case of the SHP-2 receptor tyrosine phosphatase an N-terminal SH2 domain binds and inactivates the phosphatase (PTP) domain. The pY-peptide-binding site on the N-terminal SH2 domain does not overlap with the PTP binding region. Nevertheless, pY-peptide binding causes domain dissociation and phosphatase activation. Comparative multi-nanosecond molecular dynamics simulations on the N SH2 domain in ligand-bound and free states have been performed to study the allosteric mechanism that leads to domain dissociation upon pY-peptide binding. Significant ligand-dependent differences in the conformational flexibility of regions that are involved in SH2-PTP domain association have been observed. The results support a mechanism of signal transduction where SH2-peptide binding modulates the domain flexibility and reduces its capacity to fit into the entrance of the PTP catalytic domain of SHP-2. PMID- 12542416 TI - ProML--the protein markup language for specification of protein sequences, structures and families. AB - We propose a specification language ProML for protein sequences, structures, and families based on the open XML standard. The language allows for portable, system independent, machine-parsable and human-readable representation of essential features of proteins. The language is of immediate use for several bioinformatics applications: we discuss clustering of proteins into families and the representation of the specific shared features of the respective clusters. Moreover, we use ProML for specification of data used in fold recognition bench marks exploiting experimentally derived distance constraints. PMID- 12542417 TI - Improving fold recognition of protein threading by experimental distance constraints. AB - We present a comprehensive analysis of methods for improving the fold recognition rate of the threading approach to protein structure prediction by the utilization of few additional distance constraints. The distance constraints between protein residues may be obtained by experiments such as mass spectrometry or NMR spectroscopy. We applied a post-filtering step with new scoring functions incorporating measures of constraint satisfaction to ranking lists of 123D threading alignments. The detailed analysis of the results on a small representative benchmark set show that the fold recognition rate can be improved significantly by up to 30% from about 54%-65% to 77%-84%, approaching the maximal attainable performance of 90% estimated by structural superposition alignments. This gain in performance adds about 10% to the recognition rate already achieved in our previous study with cross-link constraints only. Additional recent results on a larger benchmark set involving a confidence function for threading predictions also indicate notable improvements by our combined approach, which should be particularly valuable for rapid structure determination and validation of protein models. PMID- 12542418 TI - A hypergraph-based method for unification of existing protein structure- and sequence-families. AB - Classification of proteins is a major challenge in bioinformatics. Here an approach is presented, that unifies different existing classifications of protein structures and sequences. Protein structural domains are represented as nodes in a hypergraph. Shared memberships in sequence families result in hyperedges in the graph. The presented method partitions the hypergraph into clusters of structural domains. Each computed cluster is based on a set of shared sequence family memberships. Thus, the clusters put existing protein sequence families into the context of structural family hierarchies. Conversely, structural domains are related to their sequence family memberships, which can be used to gain further knowledge about the respective structural families. PMID- 12542419 TI - Comparing bound and unbound protein structures using energy calculation and rotamer statistics. AB - Protein data in the PDB covers only a snapshot of a protein structure. For flexible docking conformational changes need to be considered. Rotamer statistics provide the likelihood for side chain conformations, and further comparison of bound and unbound state yields differences in preferred positions. Furthermore, we do a full sampling of selected chi angles and apply the AMBER force field. Conformation of energy minima complies with the rotamer statistics. Both types of information target the reduction of search space for enumerative docking algorithms and provide parameters for elastic docking. PMID- 12542420 TI - Prediction and uncertainty in the analysis of gene expression profiles. AB - We have developed a complete statistical model for the analysis of tumor specific gene expression profiles. The approach provides investigators with a global overview on large scale gene expression data, indicating aspects of the data that relate to tumor phenotype, but also summarizing the uncertainties inherent in classification of tumor types. We demonstrate the use of this method in the context of a gene expression profiling study of 27 human breast cancers. The study is aimed at defining molecular characteristics of tumors that reflect estrogen receptor tatus. In addition to good predictive performance with respect to pure classification of the expression profiles, the model also uncovers conflicts in the data with respect to the classification of some of the tumors, highlighting them as critical cases for which additional investigations are appropriate. PMID- 12542421 TI - Impact of integrating clinical and genetic information. AB - To assess the relevance of molecular markers it is required to combine clinical and genetic information. For reliable assessment of parameters relevant to diagnostics and therapy large patient collectives must be characterized both with respect to phenotype and genotype. Matching of genetic data like gene expression profiles, molecular genetics and cytogenetics with clinical data like follow-up, morphological findings and diagnoses involves integration of complex databases. In the context of a nationwide leukemia research network in Germany we designed an integrated database covering both genetic and clinical data of patients. The system contains follow-up data and relevant laboratory modalities, i. e. cytomorphology, cytogenetics, molecular genetics, FISH, immunophenotyping and gene expression profiling. So far 13,541 cases from 7,746 patients treated by 1,225 physicians are documented. The data structure consists of up to 888 variables per case. From our experience, integration of clinical and genetic information requires significant efforts - including data protection issues -, but is feasible and improves data quality leading to faster and more reliable research results for the benefit of the patients. PMID- 12542422 TI - Modeling of self-organized avascular tumor growth with a hybrid cellular automaton. AB - Pattern formation in multicellular spheroids is addressed with a hybrid lattice gas cellular automaton model. Multicellular spheroids serve as experimental model system for the study of avascular tumor growth. Typically, multicellular spheroids consist of a necrotic core surrounded by rings of quiescent and proliferating tumor cells, respectively. Furthermore, after an initial exponential growth phase further spheroid growth is significantly slowed down even if further nutrient is supplied. The cellular automaton model explicitly takes into account mitosis, apoptosis and necrosis as well as nutrient consumption and a diffusible signal that is emitted by cells becoming necrotic. All cells follow identical interaction rules. The necrotic signal induces a chemotactic migration of tumor cells towards maximal signal concentrations. Starting from a small number of tumor cells automaton simulations exhibit the self-organized formation of a layered structure consisting of a necrotic core, a ring of quiescent tumor cells and a thin outer ring of proliferating tumor cells. PMID- 12542423 TI - Supporting genotype-phenotype correlation with the rare metabolic diseases database Ramedis. AB - To gain further knowledge about rare genetic diseases, a world wide method for data collection via the Internet has been established. This new approach will improve collecting valuable data from single case reports. Ramedis saves standardised patient data which will be usable for statistics, longitudinal examinations and cooperative studies in future time. Embedded in the scene of the German Human Genome Project, Ramedis directly will enable phenotype-genotype correlations. Beside the better characterisation of clinical heterogeneity of rare metabolic diseases, there may be a great benefit for the treatment of these patients in whom prospective studies are otherwise expensive and difficult to perform. This contribution presents the motivation for this system, introduces features, current state and the future of the project. Additionally, first experiences of using Ramedis by health professionals are explained. PMID- 12542424 TI - The system architecture of the BioPath system. AB - BioPath is a prototype system for the interactive exploration of biochemical pathways. It has been developed as an electronic version of the famous Boehringer Biochemical Pathways map and offers various ways to access information on substances and pathways and to navigate through pathways. This paper describes the main features and the software architecture of BioPath. The companion paper [11] focuses on the advanced visualization incorporated into BioPath. PMID- 12542425 TI - Rapid generation of a representative ensemble of N-glycan conformations. AB - Glycosylated proteins are ubiquitous components of extracellular matrices and cellular surfaces where their oligosaccharide moieties are implicated in a wide range of cell-cell and cell-matrix recognition events. Glycans constitute highly flexible molecules. Only a small number of glycan X-ray structures is available for which sufficient electron density for an entire oligosaccharide chain has been observed. An unambiguous structure determination based on NMR-derived geometric constraints alone is often not possible. Time consuming computational approaches such as Monte Carlo calculations and molecular dynamics simulations have been widely used to explore the conformational space accessible to complex carbohydrates. The generation of a comprehensive data base for N-glycan fragments based on long time molecular dynamics simulations is presented. The fragments are chosen in such a way that the effects of branched N-glycan structures are taken into account. The prediction database constitutes the basis of a procedure to generate a complete set of all possible conformations for a given N-glycan. The constructed conformations are ranked according to their energy content. The resulting conformations are in reasonable agreement with experimental data. A web interface has been established (http://www.dkfz.de/spec/glydict/), which enables to input any N-glycan of interest and to receive an ensemble of generated conformations within a few minutes. PMID- 12542426 TI - A hidden periodontitis epidemic during the 20th century? AB - OBJECTIVES: Increasing evidence suggests a strong causal link between smoking and periodontitis. The goal of this study was to impute how the secular changes in smoking prevalence during the 20th century impacted the advanced periodontitis incidence in the US. METHODS: Epidemiological analyses based on US prevalence data of advanced periodontitis and smoking, and predictions of future smoking prevalence. RESULTS: Assuming other risk factors for periodontitis remained constant, we estimated that the incidence of advanced periodontitis decreased by 31% between 1955 and 2000. The changes in smoking habits, and consequently the changes in periodontitis incidence, depended strongly on education and gender. Between 1966 and 1998, we estimated a 43% decreased periodontitis incidence among college-educated individuals versus only an 8% decrease among individuals with less than a high school education. Between 1955 and 1999, we estimated a 41% decrease among males versus a 14% decrease among females. By the year 2020, the incidence of advanced periodontitis may decrease 43% from its level in 1955. CONCLUSIONS: A periodontitis epidemic fueled by smoking remained hidden for most of the 20th century. Because this epidemic was hidden, it distorted our understanding of the treatment and etiology of periodontitis. The socioeconomic polarization of this epidemic will dictate alterations in patterns of periodontal care. PMID- 12542427 TI - Applicability of an abbreviated version of the oral impacts on daily performances (OIDP) scale for use among Tanzanian students. AB - OBJECTIVES: The objective was to examine the applicability of an abbreviated version of the oral impact of daily performances (OIDP) inventory to Tanzanian adults attending the University of Dar es Salaam. METHOD: A total of 1123 students (mean age 26.4 years, response rate 58%) completed a survey instrument designed to measure subjective oral health indicators including the eight-item OIDP frequency scores. After a period of 4 weeks, 228 students (mean age 24.6 years, response rate 65%) were examined clinically for the presence or absence of oral disorders. RESULTS: A total of 51% of the participants reported that an oral problem had affected them on at least one daily performance in the 6 months preceding the survey. Cronbach's alpha for the OIDP frequency items was 0.83 and 0.87 on the first and second administration of the questionnaire. A reliability coefficient (Spearman's rho) of 0.87 was obtained for the OIDP frequency scores. For five of the eight OIDP frequency items, the kappa values ranged from 0.60 to 1.0, whereas one scale item had the less satisfactory value of 0.22. Construct validity was demonstrated in that the OIDP frequency scores were statistically significantly associated in the expected direction with clinically observed oral problems and a global self-report indicator of oral health status, respectively. CONCLUSION: The OIDP frequency scale seems to have acceptable psychometric properties in the context of a descriptive questionnaire survey among Tanzanian university students. PMID- 12542428 TI - Atraumatic perspectives of ART: psychological and physiological aspects of treatment with and without rotary instruments. AB - Atraumatic Restorative Treatment, ART, is a method of minimal caries intervention that uses only hand instruments. The aim of the present study was to explore a possible difference between the extent of discomfort experienced during dental treatment according to the ART approach and a method using rotary instruments. The study was performed in Indonesia. A total of 403 children were randomly divided in two groups. In each child, one class II restoration in a deciduous molar was made. One group received treatment using rotary instruments (750 r.p.m.). The other group was treated according to the ART approach. Glass ionomer cement was used for restoration in both groups. Discomfort scores were determined using both physiological measurements (heart rate) and behavioral observations (Venham) on specific moments during the treatment. Venham scores showed a marked difference between the two groups at most time points. Heart rate measurements were different at deep excavation. Also, a clear relation between Venham scores and heart rate measurements could be found at all time points. Confounding could be shown for operating dentist, gender of the patient and initial anxiety, not for age. No effect modification could be shown. It can be concluded that children treated according to the ART approach using hand instruments alone experience less discomfort than those treated using rotary instruments. PMID- 12542429 TI - Two-year incidence of oral disadvantage, a measure of oral health-related quality of life. AB - Dental research has progressed from describing the burden of oral disease using traditional epidemiologic measures of incidence and prevalence, to measuring how oral disease, oral signs, and oral symptoms affect the daily activities and the overall quality of life of the individual. However, longitudinal evaluation of these associations remains rare. OBJECTIVES: To (i). describe the 2-year incidence and patterns of oral disadvantage; (ii). identify dimensions of oral health measures that are significant antecedents of oral disadvantage; and (iii). determine which oral health dimensions are the most strongly predictive of oral disadvantage. METHODS: The Florida Dental Care Study was a longitudinal study of oral health in diverse groups of persons who at baseline had at least one tooth and were 45 years or older. Incidence rates, odds ratios, and 95% confidence intervals were used to describe oral disadvantage and its relation to other measures of oral health. RESULTS: Nearly one-half of the participants experienced oral disadvantage at least once during 24 months of follow-up. The strongest antecedents associated with oral disadvantage were toothache pain and chewing difficulty. CONCLUSIONS: The incidence of oral disadvantage is substantial and consistent with the notion that oral health has a substantial impact on quality of life. Measures of oral pain and oral functional limitation were more strongly predictive of oral disadvantage than disease and tissue damage antecedents. PMID- 12542430 TI - The emergence of permanent teeth in Flemish children. AB - The aim of the present study was to investigate the timing of emergence of permanent teeth in Flanders (Belgium). Data were obtained in the Signal Tandmobiel projecta, a longitudinal study in a sample of 4468 Flemish children. Survival analysis with a log-logistic distribution was performed to calculate median (and mean) values and 95% CI's for all permanent teeth. The analysis indicated significantly earlier emergence ages in girls than in boys. The emergence pattern turned out to be symmetric in both sexes. Most mandibular teeth emerged significantly before their antagonists, in boys as well as in girls. These findings are in agreement with other studies on the emergence of permanent teeth. PMID- 12542431 TI - Medically administered antibiotics, dietary habits, fluoride intake and dental caries experience in the primary dentition. AB - OBJECTIVES: Dental caries continues to be a major public health problem in the US and other nations, particularly among younger children. Unfortunately, understanding of factors contributing to caries in this age group is incomplete and further studies are necessary. The purpose of this study was to assess factors related to extensive caries experience among children aged 4-7 years, including antibiotic use, dietary patterns, fluoride intakes, and tooth brushing habits. METHODS: The case-control study was nested within a longitudinal cohort study of children's oral health. The larger cohort study collected data on fluoride intakes and behavioral factors, including antibiotic use and dietary patterns of children periodically from birth. A single dental examination was completed for each child at a time between the ages of 4-7 years. A case-control design was used to compare children with severe caries experience (6 or greater dmfs) to those with no caries experience. Cases (n = 39) included all who met the criteria and had sufficient longitudinal data (n = 39) while a control group was selected to have comparable mean age, exam dates, and an equal number of boys and girls. RESULTS: Backward elimination logistic regression controlling for demographic variables showed severe caries experience to be related to greater regular pop/other sugared beverage intake, greater frequency of starch foods, and greater frequency of eating occasions. Mother's education and fluoride intake from water were inversely related to severe caries experience. Antibiotic use was generally higher in the control group, but was not significantly different than in the severe caries group. CONCLUSIONS: Although we cannot rule out the possibility that antibiotics may be associated with increased or decreased caries risk, our data suggest that they play only a minor role in caries prevention compared to fluoride. Dietary sugar continues to be a significant risk factor for caries in the primary dentition. PMID- 12542432 TI - Trauma-related phenomena in anxious dental patients. AB - OBJECTIVES: The main purpose of this descriptive study was to investigate whether dental anxiety is associated with the occurrence of trauma-related symptoms (i.e. recurrent memories and a tendency to avoid these memories) associated with earlier traumatic dental experiences. METHODS: A sample of 37 consecutive anxious patients attending a dental fear clinic was assessed prior to dental treatment. The results were compared with those of a sample of 32 consecutive patients of a general dental practice, who served as a reference group. RESULTS: The proportion of patients who indicated they had had a horrific dental experience at least once in their life did not significantly differ between the anxious group and the reference group. Conversely, significantly more patients (76%) in the anxious group reported suffering from memories of these events. They also showed a higher level of trauma-related symptomatology as indexed by the Impact of Event Scale (IES). About half of the anxious patients suffered from symptoms typically reported by patients with posttraumatic stress disorder (PTSD). Severity of dental anxiety showed a high correlation with both frequency of intrusions (r = 0.64, P < 0.001) and avoidance of the memories (r = 0.62, P < 0.001). CONCLUSIONS: The results suggest that in anticipation of treatment dentally anxious individuals suffer from a high level of intrusive recollections of earlier dental experiences. PMID- 12542433 TI - Risk indicators for apical periodontitis. AB - OBJECTIVES: The aim of the study was to identify risk indicators for apical periodontitis (AP) in the individual. A parameter was defined as a 'risk indicator' when its presence increased the individual's risk of exhibiting AP. Risk indicators may, but need not be directly associated with the disease. METHODS: The study included full-mouth surveys of 613 randomly selected individuals, who were 20-60 years old and living in Aarhus County. Logistic regression analyses were performed. The outcome variable was >or=1 AP/no AP. The independent variables were obtained from information on socioeconomic and dental status. RESULTS: It was found that smoking, no usage of services from the dentist, >or=2 secondary caries lesions, >or=3 inadequate coronal fillings and the presence of root filling(s) were statistically associated with AP. A separate analysis for individuals with no previous root fillings showed that individuals with regular dental visits as well as individuals who had all their teeth (third three molars excluded) were less likely to have AP, whereas individuals who had >or=3 inadequate coronal fillings were more likely to have AP. CONCLUSIONS: The results from the present study of a Danish population showed that the most important risk indicator of having AP in the individual was the radiographic evidence of root fillings. The presence of several caries lesions, the quality of the dental treatment, the regularity of dental visits and smoking were also statistically associated with AP. Further, the socioeconomic status of the individual did not provide much additional information on the periapical status. PMID- 12542434 TI - Exposure to fluoridated drinking water and dental caries experience in Australian army recruits, 1996. AB - OBJECTIVES: The purpose of this study was to investigate a group of young Australian adults to determine their caries experience and measure associations between caries experience and age, gender, socioeconomic status, education level and lifetime exposure to fluoridated water. METHODS: This was achieved through a cross-sectional study involving Australian Army recruits seen for their initial dental examination on enlistment into the Australian Army. A total of 499 recruits had a clinical examination with the aid of bitewing radiographs and an orthopantomograph (OPG). Sociodemographic and fluoride exposure data were obtained via a questionnaire. RESULTS: This study showed that subjects with a lifetime exposure to fluoridated water reported a 23% lower level of caries experience than subjects with no exposure to fluoridated water, with a greater effect on proximal surfaces compared to smooth and occlusal surfaces. Female subjects had a level of caries experience 25% higher than male subjects, while subjects from the lowest socioeconomic background had a level of caries experience 89% times greater than subjects from the highest socioeconomic group. CONCLUSIONS: Although it is not possible to directly establish a causal relationship from a cross-sectional study such as this, the results from this study show a dose-response relationship which suggests that there are benefits of lifetime exposure to fluoridated drinking water through young adulthood. PMID- 12542435 TI - British society for haematology slide session, annual scientific meeting, Harrogate, 2001. PMID- 12542436 TI - Significant differences between capillary and venous complete blood counts in the neonatal period. AB - The normal capillary and venous hematologic values for neonates have not been defined clearly. It is well known that capillary blood has higher hemoglobin (Hb) and hematocrit (Hct) values than venous blood. In a recent study, we reported differences between capillary and venous complete blood counts (CBC) in healthy term neonates on day 1 of life. The aim of this study was to extend our previous investigation. Term neonates (n=141) were stratified into four groups by days of postnatal age: group 2 (day 7, n=38), group 3 (day 14, n=35), group 4 (day 21, n=32) and, group 5 (day 28, n=36). Data from our previous study were included in the statistical analysis as group 1 (day 1, n=95). A CBC and differential count were carried out on each capillary and venous sample drawn simultaneously. Within each group, the mean and standard deviation for each parameter in capillary and venous blood were calculated and then compared using the paired sample t-test. In all groups, the capillary blood samples had higher Hb, Hct, red blood cell (RBC), white blood cell (WBC), and lymphocyte counts. In each group, venous platelet counts were significantly higher than the corresponding capillary values. There was also a trend toward higher venous mean corpuscular volume, higher capillary polymorphonuclear leukocyte (PML) count and mean platelet volume in all groups. In both capillary and venous blood, Hb, Hct, RBC, MCV values and WBC, lymphocyte, PML counts decreased and platelet counts increased steadily during neonatal period. This study reveals that CBC parameters and differential counts may differ depending on the blood sampling used. The findings underline the importance of considering the sample source when using hematologic reference ranges for healthy or septic neonates. When interpreting results, the term 'peripheral blood' should be replaced with 'capillary blood' or 'venous blood' so that an accurate assessment can be made. PMID- 12542437 TI - Development and clinical application of nucleated red blood cell counting and staging on the automated haematology analyser XE-2100. AB - We initially developed a new flow cytometric (FCM) reference method for the enumeration and staging of nucleated red blood cells (NRBC) in 1997 [Wang et al., 1998 (XIth International Symposium on Technological Innovations in Laboratory Haematology, Banff, Canada, 1998); Tsuji et al., 1999 (Cytometry, 1999)]. The method used CD45 antibody and propidium iodide staining to separate NRBCs from other cells. Accuracy and precision were enhanced because larger numbers of cells were counted than was possible with the manual method. We also developed a method for automated NRBC counting on a haematology analyser, the XE-2100 (Wang, 1988). NRBC were separated from other cells using a special lysing buffer and a fluorescent dye. The XE-2100 was found to detect peripheral and cord blood NRBC accurately and precisely when compared with cell morphology or FCM control methods. The FCM NRBC staging method was established through the identification of different NRBC populations following the novel staining and lysing method. To evaluate the method further, we sorted samples containing NRBCs using a FACSort and investigated NRBC staging on the Sysmex XE-2100 based on the cell sorting results. Data were analysed using special software (ida). First, we used the data in various parameter combinations. We then established gates to classify the NRBC populations. Finally, we analysed blood specimens from patients with different types of diseases to explore possible clinical applications. PMID- 12542438 TI - The utility of the autohaemolysis test for children with congenital haemolytic anaemia. AB - Autohaemolysis testing can assist the evaluation of haemolytic anaemia, but involves a cumbersome assay that is difficult to perform accurately. Because this test persists in clinical practice without clear indications or guidelines, we retrospectively reviewed our experience with autohaemolysis testing for children with suspected congenital haemolytic anaemia. Over 12 years, autohaemolysis without glucose was elevated for 38 of 39 children with congenital spherocytosis, while glucose corrected or reduced autohaemolysis in 33 of these patients. Autohaemolysis was elevated only once among seven other cases (four with congenital nonspherocytic haemolytic anaemia and three normal siblings). In three cases of congenital spherocytosis with equivocal osmotic fragility, autohaemolysis was abnormal and corrected with glucose in two. In our experience, autohaemolysis testing was helpful diagnostically in only two of 54 cases, so has only limited utility as a routine test for children with suspected congenital haemolytic anaemia. PMID- 12542439 TI - alpha-Thalassemia 2, 3.7 kb deletion and hemoglobin AC heterozygosity in pregnancy: a molecular and hematological analysis. AB - alpha-Thalassemia is a synthesis hemoglobinopathy with a worldwide distribution. alpha-thalassemia-23.7kb (alpha-Thal23.7kb) was investigated by PCR and standard hematologic analysis techniques in 106 pregnant women - 53 heterozygous for hemoglobin (Hb) A and C (AC) and 53 homozygous for the normal Hb A (AA) with similar ages and race ancestry. Eleven (21%) of AC women were alpha-Thal23.7kb heterozygous and 1 (2%) was homozygous, while 12 AA women (23%) were heterozygous. In the AA group, the MCV differed among those with normal alpha genes and those with alpha-Thal23.7kb (P = 0.031). Statistical analysis of AC group patients with normal alpha genes and alpha-Thal23.7kb carriers showed differences in MCV (P = 0.001); MCH (P = 0.003) and Hb C concentrations (P = 0.011). Analysis of AA and AC group patients with normal alpha genes showed differences in RBC (P = 0.033), Hb concentration (P = 0.003) and MCHC (P < 0.0001). There were no statistically significant differences for any hematologic parameters between AC and AA group patients with the alpha-Thal23.7kb genotype. The AC alpha-Thal23.7kb homozygous women had low hematologic parameters. Serum ferritin levels were normal among the groups studied. These results emphasize the importance of diagnosis and follow-up of patients with hemoglobinopathy carriers during pregnancy in order to administer adequate therapy and avoid further complications for mothers and newborns. PMID- 12542440 TI - The effects of high dose methylprednisolone on apoptosis in children with acute lymphoblastic leukemia. AB - Rapid leukemic cell kill at initial diagnosis of patients with acute lymphoblastic leukemia (ALL) has been shown to be associated with a favorable outcome. The aim of the present study was to investigate the effect of high dose methylprednisolone (HDMP) on in vivo blast cell apoptosis in children with ALL. Annexin V-binding and Fas (CD95), Fas ligand (FasL; CD95L), and Bcl-2 expression in PB blasts were determined in newly diagnosed children with ALL before and 4, 24, 96 h after initiation of HDMP treatment (n=20) or conventional dose steroids (CDS) (n=10) as the control group. A decrease in absolute blast count (from 40.8 x 09 to 21.4 x 109/l) associated with an increase in apoptosis (14.2 to 26.9%) (P < 0.05) was detected 4 h after initiation of HDMP. A significant increase in Fas and FasL expression was detected 96 h after HDMP. There was no significant change in apoptosis, Fas and FasL expression from baseline in the control group treated with CDS. The changes in Bcl-2 expression after treatment was not significant in both groups. The results of this preliminary study have shown that HDMP treatment was effective in inducing immediate (within 4 h) blast cell apoptosis. The contribution of Fas/FasL interaction in the rapid component of cell kill remains to be determined, as the increase in the expression of these molecules was evident later. PMID- 12542441 TI - The clinical and prognostic significance of erythrocyte sedimentation rate (ESR), serum interleukin-6 (IL-6) and acute phase protein levels in multiple myeloma. AB - Interleukin-6 (IL-6) and acute phase proteins are commonly increased in patients with multiple myeloma. Several of these acute phase proteins are believed to predict prognosis and influence survival. We measured interleukin-6 (IL-6), C reactive protein (CRP), alpha-1-antitrypsin (a1AT), acid alpha-1-glycoprotein (a1AG), haptoglobin (HAP), transferrin (TRF), hemoglobin (Hb), beta-2 microglobulin (beta2M) and erythrocyte sedimentation rate (ESR) in 42 newly diagnosed multiple myeloma patients and 25 normal controls. At the time of blood collection, nine patients were at stage I of disease, 14 at stage II, and 19 at stage III according to the Durie and Salmon myeloma staging system. Mean +/- SD values of IL-6, CRP, a1AT, a1AG, HAP, beta2M, and ESR were significantly higher and Hb significantly lower than those found in the controls. Univariate analysis, using the log-rank test, showed that among the acute phase proteins, serum CRP (P < 0.002), a1AT (P < 0.008) and ESR (P < 0.008) were significantly correlated with survival. However, when a multivariate Cox proportional hazard model was performed, ESR, CRP, a1AT, a1AG and beta2M were identified as independent prognostic factors, while the others were not. We conclude that ESR, a simple and easily performed marker, was found to be an independent prognostic factor for survival in patients with multiple myeloma. PMID- 12542442 TI - Immature reticulocytes as an early predictor of engraftment in autologous and allogeneic bone marrow transplantation. AB - The purpose of this study was to evaluate reticulocyte parameters by means of flow cytometric reticulocyte counting in a group of patients who had undergone autologous and allogeneic bone marrow transplantation (BMT). The pattern of reticulocyte response and the predictive value of absolute neutrophil count (ANC), platelet count, number of CD34+ cell infused and graft source for reticulocyte response were studied. We compared absolute reticulocyte count (RetAbs), mean fluorescence index (MFI) and mean reticulocyte volume/mean corpuscular volume (MRV/MCV) ratio with conventional criteria (ANC and platelet count) in 22 allogeneic and 20 autologous BMT recipients. An abrupt increase in MRV/MCV ratio or a rise in MFI value were the earliest signs of erythropoietic recovery following allogeneic transplantation (63.6 and 22.8% of cases, respectively). In 13.6% of the cases, both parameters were observed simultaneously. All but three autologous transplant recipients showed changes in reticulocyte parameters earlier than ANC recovery. Granulocyte recovery and peripheral blood progenitor cells (PBPC) graft were predictive variables for RetAbs response in allogeneic transplant recipients. In the autologous group, predictive variables for RetAbs response were a high number of CD34+ infused cells and platelet recovery. An increase in the immature reticulocyte population is the earliest sign of haematopoietic recovery following BMT. PMID- 12542443 TI - Consolidation therapy with autologous blood stem cell transplantation in a patient with primary plasma cell leukaemia. AB - Primary plasma cell leukaemia (PPCL) is a rare form of plasma cell dyscrasia. Conventional melphalan-based treatment is often ineffective, with a reported median survival of 2-7 months only. We report a 53-year-old man with PPCL who was treated with four cycles of combination chemotherapy including vincristine, adriamycin and dexamethasone that resulted in a good partial remission. High-dose melphalan 200 mg/m2 and autologous peripheral blood stem cell (PBSC) rescue was then given 6 months after diagnosis. Maintenance interferon-alpha was started 8 weeks after transplantation with good drug compliance. Complete remission was achieved and molecular remission was documented 11 months after autologous PBSC transplantation. In conclusion, high-dose therapy followed by autologous stem cell rescue is a feasible option for PPCL that can result in a reasonably sustained remission. PMID- 12542444 TI - A new platelet parameter, the mean platelet component, can demonstrate abnormal platelet function and structure in myelodysplasia. AB - The mean platelet component (MPC) is a new platelet parameter generated by the Bayer ADVIA 120 full blood count analyser as part of the routine complete blood count (CBC) test cycle. We report a case of myelodysplasia with bleeding complications and abnormal template bleeding time in whom low mean platelet component parameters were associated with partial platelet granule deficiency, demonstrated by transmission electron microscopy. We suggest that the mean platelet component is an inexpensive and rapid test to screen for platelet dysfunction related to ultrastructural abnormalities in myelodysplasia. PMID- 12542445 TI - Isolated thrombocytopenia: the presenting finding of typhoid fever. AB - Thrombocytopenia is generally seen as a complication in typhoid fever. However, it can also be encountered as a presenting sign on admission. A 29-year-old man with complaints of fever and diarrhoea was hospitalized because of isolated thrombocytopenia encountered on routine complete blood count examination. The diagnosis of typhoid fever was established when Salmonella typhi was isolated from the blood cultures. The platelet count returned to normal level within the first week of ceftriaxone therapy. Possible mechanisms of thrombocytopenia were discussed. PMID- 12542446 TI - Pitfalls of CD45 gating strategies in leukaemia immunophenotyping. PMID- 12542447 TI - Gender differences in the regulation of vascular tone. AB - 1. The greater incidence of hypertension and coronary artery disease in men and post-menopausal women compared with premenopausal women has suggested vascular protective effects of the female sex hormone oestrogen. However, vascular effects of the female sex hormone progesterone and the male sex hormone testosterone have also been suggested. 2. Oestrogen, progesterone and testosterone receptors have been identified in the plasmalemma, cytosol and nuclear compartments of vascular cells. The interaction of sex hormones with their specific receptors triggers not only long-term genomic vascular effects, but also acute non-genomic vascular responses. 3. Sex hormones may activate endothelium-dependent vascular relaxation pathways, including the nitric oxide-cGMP and prostacyclin-cAMP pathways and a hyperpolarizing factor pathway. 4. Sex hormones may also inhibit the mechanisms of vascular smooth muscle contraction, such as [Ca2+]i, protein kinase C and other protein kinases. 5. The sex hormone-induced stimulation of endothelium dependent vascular relaxation and inhibition of vascular smooth muscle contraction may contribute to the gender differences in vascular tone and may represent potential beneficial vascular effects of hormone-replacement therapy during natural and surgically induced deficiencies of gonadal hormones. PMID- 12542448 TI - Intracerebral microdialysis: 30 years as a tool for the neuroscientist. AB - 1. Microdialysis is an established technique for studying physiological, pharmacological and pathological changes of a wide range of low molecular weight substances in the brain extracellular fluid. Many studies have proven its sensitivity in sampling the extracellular space in discrete brain locations, such as the striatum, and monitoring the action of exogenous substances. 2. The two main areas of application of microdialysis are the recovery of endogenous substances, primarily the neurotransmitters, and the infusion of drugs through the microdialysis cannula (retrodialysis). 3. Microdialysis in awake animals is the tool of choice for studying the relationship between changes in behaviour and neurotransmitters in certain brain areas. In addition, the concomitant recording of the electroencephalogram at the site of microdialysis has been shown to be extremely useful in determining the role of certain neurotransmitters in paroxysmal activity. 4. Clinical applications of microdialysis have included monitoring of ischaemic injury, subarachnoid haemorrhage, trauma and epilepsy. With the recent availability of standardized equipment, the use of microdialysis in the neurological clinic is likely to become more common. PMID- 12542449 TI - Survival, haemodynamics and cardiac remodelling follow up in mice after myocardial infarction. AB - 1. In the present study, the time-course, over a 1 year period, of postischaemic dilated cardiomyopathy and/or development of congestive heart failure was investigated in mice in terms of survival and cardiac functional and structural characteristics. 2. C57BL/6 mice with myocardial infarction (MI mice; coronary ligation n = 78) or sham-operated animals (n = 45) were used and echocardiographic, haemodynamic and histomorphometric parameters were assessed at 3, 6 and 12 months post-MI. 3. At 12 months, the survival rate was 70% in MI mice. Left ventricular dysfunction was evidenced by a strong decrease in ejection fraction (EF; -48 and -53% at 6 and 12 months, respectively; both P < 0.05) and an increase in left ventricular end-diastolic pressure (+100% at both 6 and 12 months; both P < 0.05). There was no major worsening in cardiac function between 6 and 12 months, suggesting strong compensatory mechanisms. Cardiac remodelling was observed, characterized by strong left ventricular hypertrophy (+38 and +62% at 6 and 12 months, respectively; both P < 0.05) and dilatation (+53% at 6 months; P < 0.05), but collagen was not significantly increased. Significant correlations were found between EF (echocardiography) and dP/dtmax, between end diastolic volume (echocardiography) and left ventricular internal perimeter (histomorphometry) and between left ventricular mass (echocardiography) and weight. 4. In conclusion, despite a high survival rate, the MI mouse model displays most of the hallmarks of postischaemic dilated cardiomyopathy and/or congestive heart failure, thus affording the necessary background for the subsequent evaluation of gene manipulation and/or drug effects. In addition, two dimensional echocardiography appears to be a suitable tool for the long-term follow up of cardiac function and remodelling in this model. PMID- 12542450 TI - Renal function in adult rats subjected to prenatal dexamethasone. AB - 1. Prenatal dexamethasone leads to low birth weight and compromises organogenesis, but its effects on nephrogenesis in male and female rats have not yet been investigated extensively. Reduced renal mass may be responsible for hypertension and renal haemodynamic and morphological adjustments to maintain the glomerular filtration rate (GFR). Subsequently, these compensatory mechanisms determine glomerular sclerosis and irreversible reduction in GFR. When a high protein diet is associated with reduced renal mass, it accelerates glomerular sclerosis and the decline in renal function. The aim of the present study was to evaluate whether rats subjected to prenatal dexamethasone and a high-protein diet during growth present a premature decline in renal function. 2. The number of nephrons and renal haemodynamics were estimated in Wistar rats fed a high-protein diet (40% protein) after weaning in offspring of dams treated with either dexamethasone (0.1 mg/kg per day) or its vehicle (control; physiological solution, 0.1 mL/kg per day) during gestation. 3. At 70 days of age, rat offspring were anaesthetized and prepared surgically for renal haemodynamic measurements. 4. Mean arterial pressure (MAP), renal blood flow (RBF) and GFR were measured using a blood pressure transducer, a flow probe and inulin clearance, respectively. 5. The number of nephrons was counted using the acid maceration technique. 6. Dexamethasone during pregnancy induced a lower weight gain in the dams (65%; P < 0.0001) and a lower birth weight in both male and female offspring (14 and 13%, respectively; P < 0.01). 7. Compared with control, the number of nephrons in male rats was reduced by 13% (30 703 +/- 1262 vs 26 308 +/- 1305, respectively; P < 0.05), but was unaltered in female rats (23 197 +/- 553 vs 24 231 +/- 1009, respectively). 8. Male and female rats did not show any alteration in MAP. In addition, they did not show any alteration in renal vascular resistance, RBF, filtration fraction or GFR. 9. In conclusion, prenatally administered dexamethasone (0.1 mg/kg during the entire pregnancy) induced a low birth weight. The magnitude of the reduction in nephrogenesis in male offspring from mothers treated with dexamethasone was not sufficient to alter renal function (measured at 70 days), even when rats had been fed a high protein diet. PMID- 12542451 TI - Antihyperglycaemic effect of Cassia auriculata in experimental diabetes and its effects on key metabolic enzymes involved in carbohydrate metabolism. AB - 1. In experimental diabetes, enzymes of glucose and fatty acid metabolism are markedly altered. Persistent hyperglycaemia is a major contributor to such metabolic alterations, which lead to the pathogenesis of diabetic complications. To our knowledge, there are no available reports on the enzymes of hepatic glucose metabolism of Cassia auriculata flower against diabetes. The present study was designed to study the effect of Cassia auriculata flower extract (CFEt) on hepatic glycolytic and gluconeogenic enzymes. 2. Streptozotocin diabetic rats were given CFEt (0.15, 0.30 and 0.45 g/kg) or 600 microg/kg glibenclamide for 30 days. At the end of 30 days, blood glucose, plasma insulin, haemoglobin, glycosylated haemoglobin, glycolytic and gluconeogenic enzymes were assessed. 3. Administration of CFEt at 0.45 g/kg significantly decreased blood glucose, glycosylated haemoglobin and gluconeogenic enzymes and increased plasma insulin, haemoglobin and hexokinase activity. Similarly, administration of glibenclamide showed a significant effect; however, CFEt at 0.15 and 0.30 g/kg did not show any significant effect. 4. In conclusion, the observations show that the aqueous extract of CFEt possesses an antihyperglycaemic effect and suggest that enhanced gluconeogenesis during diabetes is shifted towards normal and that the extract enhances the utilization of glucose through increased glycolysis. The effect of CFEt was more prominent than that of glibenclamide. PMID- 12542452 TI - Effect of Undaria pinnatifida (Wakame) on the development of cerebrovascular diseases in stroke-prone spontaneously hypertensive rats. AB - 1. We showed that a nutritional factor was able to attenuate the development of hypertension and its related diseases in stroke-prone spontaneously hypertensive rats (SHRSP). In the present study, the effect of Wakame, an edible brown seaweed, on the development of stroke was examined in SHRSP. 2. We studied the treatment with 5% (w/w in a diet) Wakame powder in salt-loaded (0.5% NaCl in drinking water) SHRSP. Salt-loaded animals treated with 5% cellulose or kaolin were used as controls. Wakame significantly delayed the development of stroke signs (P < 0.05) and significantly improved the survival rate of salt-loaded SHRSP (P < 0.05). There was no significant difference in the elevation of blood pressure among the three groups during the observation period. 3. We isolated fucoxanthin, a carotinoid, from Wakame powder and studied its preventive effect on ischaemic cultured neuronal cell death. Fucoxanthin significantly attenuated neuronal cell injury in hypoxia and re-oxygenation (P < 0.05). 4. Based on these results, we conclude that Wakame has a beneficial effect on cerebrovascular diseases in SHRSP, independent of hypertension. It is possible that fucoxanthin in Wakame may have a preventive effect against ischaemic neuronal cell death seen in SHRSP with stroke. PMID- 12542453 TI - Restoration of arterial baroreflex function contributes to organ protection in spontaneously hypertensive rats treated with long-term hydrochlorothiazide mixture. AB - 1. Hydrochlorothiazide mixture (HCTM) is widely used in China for the treatment of hypertension. This mixture consists of hydrochlorothiazide, triamterene, reserpine, hydralazine and chlordiazpoxide, with small (one-third to one-fifth of normal) doses of each drug. The present study was designed to investigate the effects of this mixture on blood pressure, blood pressure variability (BPV), baroreflex sensitivity (BRS) and end-organ damage in spontaneously hypertensive rats (SHR). 2. The HCTM was mixed in the rat chow and rats were treated for 4 months. After treatment, rats were catheterized and their blood pressure, BPV and BRS were measured in the conscious state. Organ damage was examined after these measurements had been completed. 3. It was found that HCTM not only decreased blood pressure and BPV, but also ameliorated impaired BRS in SHR. The HCTM had an obvious effect on organ protection in SHR. 4. The HCTM prevented left ventricular hypertrophy and this effect was mainly related to a decrease in systolic blood pressure. The effects of HCTM on preventing renal atrophy were mainly determined by BRS. Baroreflex sensitivity was the most important determinant for predicting organ damage in HCTM-treated SHR. 5. In conclusion, long-term treatment of rats with HCTM prevented hypertensive organ damage. Restoration of arterial baroreflex function contributes to organ protection in SHR treated in the long term with HCTM. PMID- 12542454 TI - Effect of 6-hydroxydopamine on gluconeogenesis in the rat renal cortex. AB - 1. In the present study, the effects of 6-hydroxydopamine (6-OHDA) on renal gluconeogenesis were investigated in vitro using rat renal cortical slices. Cisplatin, a known nephrotoxin, was used as a positive control. The working hypothesis for the present study was that 6-OHDA, as a reactive oxygen species producing agent, could inhibit renal gluconeogenesis. 2. 6-Hydroxydopamine is used for chemical sympathectomy because it selectively destroys adrenergic nerve endings. Long-term use of levodopa causes a variety of side-effects in parkinsonian patients. 6-Hydroxydopamine has been reported to be present in the urine of parkinsonian patients on levodopa medication. The renal toxicity of endogenously formed 6-OHDA is a matter of concern in these patients. 3. In one series of experiments, renal cortical slices were incubated for 60 min in medium containing 0.5, 1.0, 2.08, 5.15, 10.30 or 20.60 mg/mL 6-OHDA at 37 degrees C under a 100% O2 atmosphere. In another series of experiments, renal cortical slices were incubated in medium containing 10.30 mg/mL 6-OHDA for 15, 30, 45, 60, 90 or 120 min or in 6-OHDA-free medium. 4. In a second series of experiments, renal cortical slices were incubated for 60 min in medium containing 0.25, 0.50, 0.75, 1.0, 1.25 or 1.50 mg/mL cisplatin at 37 degrees C under a 100% O2 atmosphere. In another set of experiments, renal cortical slices were incubated in medium containing 1 mg/mL cisplatin for 15, 30, 45, 60, 90 or 120 min or in a cisplatin-free medium. 5. The results of the studies in which slices were incubated in 6-OHDA-containing media indicate that 6-OHDA induced a time- and concentration-dependent decrease in renal gluconeogenesis. Therefore, 6-OHDA causes functional injury of renal proximal tubule cells responsible for renal gluconeogenesis, thus leading to nephrotoxicity. PMID- 12542455 TI - Effects of second-generation histamine H1 receptor antagonists on the active avoidance response in rats. AB - 1. The aim of the present study was to establish a new schedule of active avoidance response in rats to estimate the central effects of second-generation histamine H1 receptor antagonists. 2. With the new schedule, a rat was placed into a dark room. A sliding door was opened after a delay of 5 s and, unless the animal moved into the lit room, an electric shock was delivered for 3 s. With the conventional schedule, the sliding door was opened immediately after the rat was placed into the dark room. 3. Ketotifen, at a dose of 50 mg/kg, showed no significant effect on the retrieval of active avoidance response with the conventional schedule. However, with the new schedule, the drug caused significant inhibition of retrieval of the response, even at a dose of 10 mg/kg. 4. Epinastine showed no significant effect on retrieval of the active avoidance response, even at a dose of 50 mg/kg with the new schedule. 5. Cetirizine, at a dose of 50 mg/kg, caused a significant effect, indicating that cetirizine, at this dose, markedly inhibits memory retrieval. 6. Both olopatadine and loratadine had potent effects; at doses of 20 and 50 mg/kg, respectively, these agents showed significant inhibitory effects on retrieval of the response. 7. In conclusion, we have developed a new schedule of active avoidance response that can be used to estimate the central effects of second-generation histamine H1 receptor antagonists. PMID- 12542456 TI - Effects of polarizing solution on glucose uptake of rat oxygenated or hypoxic ventricular myocytes. AB - 1. Although adult cardiac metabolism depends on fatty acids rather than on carbohydrates, hypoxic hearts consume glucose and infusion with a glucose-insulin potassium solution (GIK) helps in the recovery of the pressure register, the heart rate and electrocardiogram of rat hypoxic whole hearts. 2. Rat myocytes obtained by enzymatic disaggregation captured significant amounts of glucose in Tyrode's solution (9.8 +/- 2.05% during oxygenation and 16.26 +/- 2.76% during hypoxia) after 60 min incubation. Cells incubated with GIK showed a significantly increased glucose uptake (to 30.04 +/- 1.99% during oxygenation and to 37.06 +/- 1.97% during hypoxia). 3. Glucose was the component of the Tyrode's solution that most enhanced its own entry, increasing glucose entry to 45.42 +/- 2.64% with oxygen and 41.42 +/- 2.69% under hypoxia. 4. Antibodies against glucose transporters 1 and 4 (GLUT-1 and -4, respectively) decreased glucose transport during GIK treatment under hypoxia. 5. We conclude that GIK has beneficial effects mediated by GLUT-1 and -4 during hypoxia, inducing recovery of the damaged tissue. PMID- 12542457 TI - Angiotensin-converting enzyme inhibition in adult hypertensive rats: a stereological study of renal filtration surface area. AB - 1. Angiotensin-converting enzyme (ACE) inhibitor treatment leads to beneficial effects on kidney function. The aim of the present study was to determine whether ACE inhibition at high or low doses affects glomerular capillary surface area and length, glomerular number or total renal filtration surface area in rats with established hypertension and, if so, to determine whether these effects are mediated through bradykinin potentiation. 2. Spontaneously hypertensive rats (SHR) were treated with the ACE inhibitor perindopril at either 3 or 0.1 mg/kg per day (high and low doses, respectively) from 16 to 24 weeks of age. Some rats were concomitantly treated with the bradykinin B2 receptor antagonist S16118 (10 nmol/kg per day). Blood pressure was measured twice weekly during the treatment period. At 24 weeks of age, rats were perfusion fixed at 140 mmHg, the kidneys removed, embedded in resin and examined stereologically to estimate glomerular number and volume, length and surface area of glomerular capillaries and total renal filtration surface area. 3. High- and low-perindopril treatment significantly reduced systolic blood pressure compared with control SHR. However, the rats treated with low-dose perindopril were still considered hypertensive. Neither low-dose nor high-dose perindopril treatment had any observable effect on glomerular number (23 876 +/- 1201 vs 26 240 +/- 1465 glomeruli/kidney, respectively) or volume (2.25 +/- 0.21 and 1.96 +/- 0.06 x 10-3 mm3, respectively) compared with controls (glomerular number 25866 +/- 1210 glomeruli/kidney; glomerular volume 2.24 +/- 0.21 x 10-3 mm3). As a result, there was no significant difference in total renal filtration surface area between any of the experimental groups (8161.6 +/- 550.9, 8699.7 +/- 427.6, 9081.9 +/- 453.6, 8830.2 +/- 521.2 and 8559.4 +/- 341.4 mm2 for SHR, SHR low-dose perindopril, SHR low-dose perindopril + B2 antagonist, SHR high-dose perindopril and SHR high-dose perindopril + B2 antagonist, respectively). Coadministration of the bradykinin antagonist had no observable effect on any of the parameters studied. 4. In conclusion, because neither high-dose nor low-dose perindopril had any effect on total renal filtration surface area, the observed beneficial effects of ACE inhibition on kidney function are not the result of enhancement in glomerular capillary surface area. PMID- 12542458 TI - Endurance training adaptations modulate the redox-force relationship of rat isolated slow-twitch skeletal muscles. AB - 1. Studies have shown that, in isolated skeletal muscles, maximum isometric force production (Po) is dependent on muscle redox state. Endurance training increases the anti-oxidant capacity of skeletal muscles, a factor that could impact on the force-producing capacity following exogenous exposure to an oxidant. We tested the hypothesis that 12 weeks treadmill training would increase anti-oxidant capacity in rat skeletal muscles and alter their response to exogenous oxidant exposure. 2. At the conclusion of the 12 week endurance-training programme, soleus (slow-twitch) muscles from trained rats had greater citrate synthase (CS) and catalase (CAT) activity compared with soleus muscles from untrained rats (P < 0.05). In contrast, CAT activity of extensor digitorum longus (EDL; fast-twitch) muscles from trained rats was not different to EDL muscles of untrained rats. The CS activity was lower in EDL muscles from trained compared with untrained rats (P < 0.05). 3. Equilibration with exogenous hydrogen peroxide (H2O2, 5 mmol/L) increased the Po of soleus muscles from untrained rats for the duration of treatment (30 min), whereas the Po of EDL muscles was affected biphasically, with a small increase initially (after 5 min), followed by a more marked decrease in Po (after 30 min). The H2O2-induced increase in Po of soleus muscles from trained rats was less than that in untrained rats (P < 0.05), but no differences were observed in the Po of EDL muscles following training. 4. The results indicate that 12 weeks endurance running training conferred adaptations in soleus but not EDL muscles. These adaptations were associated with an attenuation of the oxidant induced increase in Po of soleus muscles from trained compared with untrained rats. We conclude that endurance training-adapted soleus muscles have a slightly altered redox-force relationship. PMID- 12542459 TI - Cardiovascular reflex responses after intrathecal omega-conotoxins or dexmedetomidine in the rabbit. AB - 1. The effects of thoracic intrathecal doses (1 microg/kg) of the alpha2 adrenoceptor agonist dexmedetomidine and omega-conotoxins MVIIA and CVID on vasoconstrictor and heart rate responses to acute central hypovolaemia were studied in seven chronically instrumented rabbits. 2. Gradual inflation of an inferior vena cava cuff to reduce cardiac index (CI) by 8% per minute induced progressive vasoconstriction and an increase in heart rate (phase I). At approximately 40% of resting CI, there was sudden decompensation with failure of vasoconstriction and decrease in mean arterial pressure (MAP; phase II). 3. Both intrathecal MVIIA and CVID decreased resting CI (by 20% at 3 h), but only MVIIA significantly reduced resting MAP (P = 0.003). Dexmedetomidine resulted in transient bradycardia, but no other significant change in the resting circulation. With simulated haemorrhage, the relationship between CI and vascular conductance was shifted after MVIIA (1-3 h after injection) so that there was less vasoconstriction and a reduced increase in heart rate by the end of phase I compared with other treatments (P = 0.002 and P = 0.009, respectively). One hour after injection, dexmedetomidine reduced the slope of the phase I vasoconstrictor response (P = 0.03), but did not significantly alter the end-point of the response. With failure of vasoconstriction and the onset of phase II, vascular conductance was higher after MVIIA compared with controls. Both conotoxins caused progressive failure of vasoconstriction rather than recovery during phase II (P < 0.001). 4. Intrathecal injections of these drugs to control chronic pain may compromise cardiovascular responses to changes in central blood volume. At the single doses studied, there were significant differences between the responses to simulated haemorrhage after MVIIA or dexmedetomidine compared with CVID, with the prolonged effect after MVIIA most likely to be of clinical significance. PMID- 12542460 TI - Green tea catechins evoke a phasic contraction in rat aorta via H2O2-mediated multiple-signalling pathways. AB - 1. The contractile effects of tea polyphenols (TP) and its four principle catechins, namely (-)-epicatechin (EC), (-)-epicatechin-3-gallate (ECG), (-) epigallocatechin (EGC) and (-)-epigallocatechin-3-gallate (EGCG), on rat aorta contractility were investigated using the isometric tension recording technique. 2. At concentrations of 5-100 mg/L, TP evoked phasic contraction of rat aorta in a concentration-dependent but endothelium-independent manner. Of the four catechins tested, EGCG and EGC (3-300 micromol/L), but not EC and ECG, mimicked the contractile response to TP, suggesting that the epigallol moiety in the B ring may be associated with the contractile effect. 3. Contractions in response to EGCG and EGC were not affected by several endogenous vasoconstrictor receptor antagonists, but could be abolished by 10 micro mol/L BAPTA-AM, a membrane permeable Ca2+ chelator, or attenuated by removal of extracellular Ca2+, suggesting the involvement of both intracellular and extracellular Ca2+ in evoking the contraction. 4. Pretreatment with non-selective Ca2+ channel antagonists mefenamic acid (10 micro mol/L), tetrandrine (30 micro mol/L) and SKF 96365 (30 micromol/L), but not nifedipine (1 micromol/L), the selective inhibitor of voltage-dependent Ca2+ channels, inhibited the contractile responses to EGC and EGCG, indicating the involvement of Ca2+ influx via non-voltage dependent Ca2+ channels. 5. Several intracellular Ca2+ channel modulators, including procaine (5 mmol/L), dantrolene (30 micromol/L) and 2-amino ethoxydiphenyl borate (50 micromol/L; an inositol 1,4,5-trisphosphate receptor inhibitor), also inhibited EGCG- and EGC-induced contractions, thus suggesting a role of intracellular Ca2+ release in these contractions. 6. Both EGCG- and EGC-induced contractions were depressed, to different degrees, by inhibitors of several receptor-coupled enzymes, including phospholipase C, protein kinase C, phospholipase A2 and tyrosine kinase. Furthermore, both EGCG- and EGC-induced contractions were completely abolished by catalase, but not by superoxide dismutase or mannitol/dimethyl sulphoxide. 7. Taken together, these data show, for the first time, that TP and its related catechins that contain an epigallol structure in the B ring, as in EGCG and EGC, exert direct contractile effects on rat aortic smooth muscle via a H2O2-mediated pathway. PMID- 12542461 TI - RNA interference may be more potent than antisense RNA in human cancer cell lines. AB - 1. RNA interference (RNAi) is a newly discovered cellular pathway for the silencing of sequence-specific genes at the mRNA level by the introduction of the cognate double-stranded (ds) RNA. Because antisense (AS) mechanisms have similar effects, we compared these two effects in human cancer cell lines, considering a possible application of RNAi for cancer therapy. 2. We tested RNAi effects by transfecting human hepatoma and pancreatic cancer cell lines with AS and sense (S) RNA expression plasmids corresponding to the exogenous luciferase gene or the endogenous c-raf gene in the form of complexes with a cationic lipopolyamine or a tumour-targeting peptide vector we developed. In addition, we compared the effects of small interfering RNA and AS oligoDNA complexed with the peptide vector. 3. From the viewpoint of AS actions, the effect of the AS RNA may be cancelled by the S RNA, although, interestingly, we found that the combination of the AS and S RNA expression plasmids was more effective than the AS RNA expression plasmids alone in reducing target gene expression, whereas the S RNA expression plasmids had no effects. The combination of the luciferase AS and S RNA had no effects on the expression of either the beta-galactosidase gene or the c-raf gene. In the presence of 2-aminopurine (an inhibitor of dsRNA-activated protein kinase), the inhibitory effect of the combination of AS and S RNA on gene expression did not change in the case of the endogenous c-raf gene, but was reduced in the case of the exogenous luciferase gene. The effect of 22 nucleotide RNA duplexes corresponding to the luciferase gene was by one order stronger than that of the phosphorothioate AS DNA. 4. Thus, it is suggested that RNAi may be more potent than AS RNA in reducing target gene expression in human cancer cell lines, regardless of the length of dsRNA. With further studies on the RNAi phenomenon in cancer cells, RNAi could provide a novel approach for cancer gene therapy. PMID- 12542462 TI - Extracellular Ca2+ is obligatory for ouabain-induced potentiation of cardiac basal energy expenditure. AB - 1. The method of action of cardiac glycosides is commonly explained by the 'pump inhibition hypothesis': inhibition of the Na+/K+-ATPase allows [Na+]i to rise, eventually reversing Na+/Ca2+ exchange. The resulting influx of Ca2+o increases [Ca2+]i, thereby activating intracellular Ca2+-dependent ATPases and, hence, energy demand. This sequence has been presumed to occur during diastole as well as systole. However, it has been reported that dihydro-ouabain-induced potentiation of heat production by quiescent ventricular trabeculae persists in the absence of Ca2+o. This implies that the pump-inhibition hypothesis is inapplicable during diastole. 2. We tested this implication by: (i). measuring the rate of oxygen consumption (Vo2) of arrested guinea-pig whole-hearts; (ii). measuring[Ca2+]i in quiescent ventricular trabeculae; and (iii). mathematical modelling using software (Oxsoft Heart, Oxford Software, Oxford, UK) based on DiFrancesco-Noble formalism. 3. Upon induction of arrest, whole heart Vo2 fell to one-quarter of its 'beating' value. Subsequent perfusion with ouabain (20 micromol/L), in the presence of Ca2+o, increased Vo2 fourfold. This increase was prevented by withholding Ca2+o. Comparable results were obtained in quiescent trabeculae: ouabain increased [Ca2+]i only if Ca2+o was present. Mathematical modelling readily simulated these experimental results. 4. We conclude that influx of Ca2+o is mandatory for potentiation of cardiac basal metabolism by cardiac glycosides. PMID- 12542463 TI - Cardiac mitochondrial complex activity is enhanced by heat shock proteins. AB - 1. Prolonged ischaemia and reperfusion in heart transplantation results in mitochondrial dysfunction and loss of cardio-energetics. Improved myocardial tolerance to ischaemia-reperfusion can be increased by de novo synthesis of heat shock protein (Hsp) groups, transiently expressed following mild hyperthermic or oxidative stress. Consideration of the roles of various Hsp in ischaemic reperfused myocardium can provide new insights into potential therapeutic adjuncts to cardiac surgery. 2. Several Hsp classes have been located within or in association with mitochondrial elements. Cardiac Hsp research has focused primarily on the 70 kDa group, involved in protein folding functions within the cytosol and matrix. Similarly, Hsp 60 and 10 have been shown to form a mitochondrial chaperonin complex conferring protection to ischaemia-challenged myocytes. Equally pertinent is Hsp 32, an isoform of the haem-metabolizing enzyme heme oxygenase. 3. Our studies have shown that mitochondrial respiratory enzyme activity can be protected by Hsp, affording protection to cardiac energetics during preservation for transplantation. Upregulation of Hsp 32, 60 and 72 in rats, achieved by mild hyperthermic stress, improved cardiac function, ultrastructure and mitochondrial respiratory and complex activities in ex vivo perfused hearts subjected to cold cardioplegic arrest and ischaemia-reperfusion. Pre-ischaemic mitochondrial complex activities were increased in heat stress versus sham-treated groups for complex I, IV and V. 4. Investigation of the direct effect of upregulation of Hsp 72 by gene transfection resulted in a similar pattern of response, with increased complex I activity and improved ventricular function. 5. These studies provide the first evidence of Hsp-mediated enhancement of mitochondrial energetic capacity. Enhanced protection of mitochondrial energetics, as a result of increased Hsp expression, contributes to the recovery of myocardial function in ischaemia-reperfusion. PMID- 12542464 TI - Why government and charitable research funding agencies should not seek commercial control or returns from the research they fund. PMID- 12542466 TI - Insect immunity and its implication in mosquito-malaria interactions. AB - Insects' resistance to infectious agents is essential for their own survival and also for the health of the plant, animal and human populations with which they closely interact. Several of the major human diseases are spread by insects and are rapidly expanding as a result of the development of insecticide resistance in vectors and drug resistance in parasites. A vector insects' permissiveness to a pathogen, and hence the spread of the disease, will largely depend on the compatibility of the molecular interactions between the two species and the capability of the insect immune system to recognize and kill the pathogen. The innate immune system comprises a variety of components and mechanisms that can discriminate between different microorganisms and mount specific responses to control pathogenic infections. An impressive body of knowledge on the insects' innate immunity has been generated from studies in the model organism Drosophila. These studies are now guiding the exploration of the immune system in the vector mosquito of human malaria, Anopheles, and its implication in the elimination of parasites. Anopheles immune responses have been linked to parasite losses and some refractory mosquitoes can kill all parasites through specific defence mechanisms. The recently sequenced Drosophila and Anopheles genomes provide a detailed and comparative view on their immune gene repertoires that in combination with post-genomic analyses is used to further dissect the complex mechanisms of Plasmodium killing in the mosquito. PMID- 12542467 TI - Anthrax toxins and the host: a story of intimacy. AB - Although the dramatic events of the year 2001 have revitalized the interest in anthrax, research on Bacillus anthracis and its major virulence factors is one of the oldest theme in microbiology and started with the early works of Robert Koch and Louis Pasteur. The anthrax toxins are central to anthrax pathogenesis. They were discovered in the mid-1950s and since then there has been an enormous amount of work to elucidate both the molecular and physiopathological details of their mode of action. In this review, after a brief introduction of B. anthracis, we will focus on the latest findings that concern two aspects of anthrax toxin research: the environmental signals and the molecular mechanisms that regulate toxin synthesis, and the mechanisms of intoxication. We hope to convince the reader that the anthrax toxins are highly specialized determinants of B. anthracis pathogenicity: their synthesis is integrated within a global virulence programme and they target key eukaryotic cell proteins. We conclude with a consideration of the therapeutic perspectives arising from our current knowledge of how the toxins work. PMID- 12542468 TI - Helicobacter pylori strains expressing the vacuolating cytotoxin interrupt phagosome maturation in macrophages by recruiting and retaining TACO (coronin 1) protein. AB - Recent evidence suggests that persistence of Helicobacter pylori can be explained, at least in part, by the failure of macrophages to kill bacteria. The fate of type 1 H. pylori strain LC11, which expresses the cag pathogenicity island (PAI) and the vacuolating cytotoxin, and type 2 strain LC20, which lacks both these virulence factors, was determined following infection of the murine macrophage cell line RAW 264.7 or the human macrophage-like cell line THP-1. Helicobacter pylori strain LC11 displayed enhanced survival in macrophages in comparison with strain LC20 (4.0 +/- 0.2 versus 2.1 +/- 0.6 log CFU ml-1, P < 0.01) at 24 h. Phagosomes containing strain LC11 showed reduced co-localization with LysoTracker Red, higher levels of expression of the early endosome marker EEA1 expression and lower expression of the late endosome/lysosome marker LAMP1 relative to internalized strain LC20, both at 2 h and 24 h. These findings indicate that, in contrast to strain LC20, strain LC11 resides in a compartment with early endosome properties and does not fuse with lysosomes. In addition, phagosomes containing LC11 recruited and retained a higher percentage of TACO (coronin 1) protein in comparison with phagosomes containing strain LC20. Furthermore, IFN-gamma stimulation facilitated maturation of phagosomes containing strain LC11 in association with the release of TACO and a reduction in bacterial survival. We have demonstrated through the use of isogenic cagA-, cagE /picB- and vacA- mutant strains, that VacA plays a significant role in the interruption of the phagosome maturation. Taken together, these results indicate that, following phagocytosis, H. pylori strains expressing the vacuolating cytotoxin arrest phagosome maturation in association with the retention of TACO. PMID- 12542469 TI - Francisella tularensis inhibits Toll-like receptor-mediated activation of intracellular signalling and secretion of TNF-alpha and IL-1 from murine macrophages. AB - Microbial ligands, including lipopolysaccharide (LPS) and bacterial lipoproteins, activate Toll-like receptors (TLR) of mononuclear phagocytes, thereby inducing proinflammatory cytokines and antimicrobial activity. We show that Francisella tularensis, an intracellular pathogen, is capable of inhibiting this macrophage response. Infection with the live vaccine strain F. tularensis LVS rendered cells of the murine macrophage-like cell line J774A.1 incapable of secreting TNF-alpha or IL-1beta and mobilizing an antimicrobial activity in response to bacterial lipopeptide or Escherichia coli-derived LPS. Inhibition of TNF-alpha secretion occurred also when J774 cells were infected with F. tularensis LVS in the presence of chloramphenicol, but not when they were infected with a mutant of F. tularensis LVS defective in expression of a 23 kDa protein that is upregulated during intracellular infection. Purified F. tularensis LPS did not show an agonistic or antagonistic effect on the E. coli LPS-induced activation of the J774 cells. Francisella tularensis LVS suppressed the capability of the cells to respond to LPS or bacterial lipopeptide (BLP) with activation of nuclear factor kappa B (NF-kappaB), and degradation of the in-hibitor of NF-kappaB, IkappaB, was blocked during the infection. Also the LPS- or BLP-induced phosphorylation of the mitogen-activated protein kinase p38 and the transcription factor c-Jun was inhibited by F. tularensis LVS but not by the 23 kDa protein mutant. In conclusion, F. tularensis appears capable of abrogating the TNF-alpha and IL-1 responses of macrophages induced by E. coli LPS or BLP via a mechanism that involves suppression of several intracellular pathways and is dependent on expression of a bacterial 23 kDa protein. PMID- 12542470 TI - Interaction between the Yersinia protein tyrosine phosphatase YopH and eukaryotic Cas/Fyb is an important virulence mechanism. AB - The tyrosine phosphatase YopH is an essential virulence factor produced by pathogenic Yersinia species. YopH is translocated into host cells via a type III secretion system and its dephosphorylating activity causes disruption of focal complex structures and blockage of the phagocytic process. Among the host cell targets of YopH are the focal adhesion proteins Crk-associated substrate (p130Cas) and focal adhesion kinase (FAK) in epithelial cells, and p130Cas and Fyn-binding protein (Fyb) in macrophages. Previous studies have shown that the N terminal domain of YopH acts as a substrate-binding domain. In this study, the mechanism and biological importance of the targeting of YopH to focal complexes relative to its interaction with p130Cas/Fyb was elucidated. Mutants of YopH that were defective in p130Cas/Fyb binding but otherwise indistinguishable from wild type were constructed. Mutants unable to bind p130Cas did not localize to focal complex structures in infected cells, indicating that the association with p130Cas is critical for appropriate subcellular localization of YopH. These yopH mutants were also clearly attenuated in virulence, showing that binding to p130Cas and/or Fyb is biologically relevant in Yersinia infections. PMID- 12542471 TI - The allele encoding the mycobacterial Erp protein affects lung disease in mice. AB - Erp (exported repetitive protein), also known as P36, Pirg and Rv3810, is a member of a mycobacteria-specific family of extracellular proteins. These proteins consist of three domains, the N- and C-terminal domains are similar in all mycobacterial species, however, the central domain contains a repeated PGLTS module and differs considerably between species. The erp knockout mutant of Mycobacterium tuberculosis displays very low levels of multiplication both in macrophage cell lines and in vivo in a mouse model of infection. The high interspecies variability of the central repeated region of the Erp protein led us to investigate whether these orthologous proteins were functionally equivalent in a mouse model of tuberculosis. We expressed a gene fusion with the erp gene of Mycobacterium smegmatis, Mycobacterium leprae or M. tuberculosis in trans in an erp-M. tuberculosis mutant and found that these three alleles restored multiplication to similar levels in the spleen of infected mice. However, these alleles gave different levels of colonization in the lung, for the early time points. Quantitative histological analyses of the lungs of infected animals showed that the nature of the erp allele strongly affected the number and the size of lung lesions, demonstrating the importance of surface determinants for virulence and tissue damage. PMID- 12542472 TI - Candidal infection diagnosed on trephine biopsy sections. PMID- 12542473 TI - Granulocytic sarcoma (chloroma) of the lung. PMID- 12542474 TI - The role of hydroxyurea in sickle cell disease. PMID- 12542475 TI - Guidelines for the diagnosis and therapy of adult myelodysplastic syndromes. PMID- 12542476 TI - Defective expression of the T-cell receptor-CD3 zeta chain in T-cell acute lymphoblastic leukaemia. AB - This study analysed the T-cell receptor (TCR)-CD3 zeta complex and the signal transduction apparatus of T-acute lymphoblastic leukaemia (T-ALL) blasts, and investigated the function of the ubiquitin-proteasome system. In all nine T-ALL samples studied, the leukaemic cells showed a marked reduction in the expression of the zeta chain, while a variety of tyrosine kinases (p56lck, ZAP70 and SYK) were normally present. There was no expression of the FcepsilonRIgamma chain. To confirm that this aberration was specific to immature T-ALL blasts, we investigated two patients with lymphoproliferative disorders of granular lymphocytes (LDGL), characterized by the expansion of mature T lymphocytes and found normal zeta chain expression. The reduction of the zeta chain protein was not reversible after 72 h stimulation with the anti-CD3 monoclonal antibody and interleukin 2, either alone or in combination. Northern blot analysis indicated that the reduced protein expression did not correspond to a defect at the mRNA level, nor were mutations in the coding region of the zeta chain found. We, therefore, hypothesized that the observed reduction of protein expression in T ALL blasts could be secondary to an increased degradation at the proteasome level. Following selective inhibition of the proteasome, a marked increase of the zeta chain expression was observed. Moreover, an increase in the surface expression of CD3 was also documented. Taken together, these results indicate that the expression of the zeta subunit of the TCR-CD3 complex is consistently reduced in T-ALL blasts and that degradation of the protein is mediated by the proteasome system. PMID- 12542477 TI - Elderly haematological patients with chemotherapy-induced febrile neutropenia have similar rates of infection and outcome to younger adults: a prospective study of risk-adapted therapy. AB - We prospectively evaluated 131 consecutive episodes of fever and chemotherapy induced neutropenia in 85 adults with haematological malignancies to determine whether older patients (aged < 60 years) have different causes of fever and outcome than younger adults (aged < 60 years). Patients were stratified into high risk and low-risk groups according to previously published criteria. High-risk patients received ceftazidime plus amikacin and low-risk patients received ceftazidime alone. All patients were hospitalized until fever and neutropenia resolved. Ninety one high-risk episodes were documented: 56 occurring in older patients (mean age 69 years) and 35 in younger adults (mean age 45 years). Non Hodkgin's lymphoma and acute myeloid leukaemia were the most frequent underlying neoplasias in both age groups. Intensity of chemotherapy was similar in both age groups. Mean neutrophil count at entry, median duration of neutropenia, rate of documented infection, incidence of bacteraemia, response to therapy, overall mortality and infectious mortality were similar in the two high-risk age subgroups. The elderly subgroup had a trend to have more Gram-negative infections and the younger patients more Gram-positive infections. In addition, 40 low-risk episodes were registered: 29 in elderly patients (mean age 68 years) and 11 in younger patients (mean age 44 years). Elderly low-risk patients had more concurrent diseases that younger ones (P = 0.124). Mean neutrophil count at entry, median duration of severe neutropenia and rate of response were similar in the two age subgroups. All low-risk patients survived. In conclusion, elderly haematological cancer patients with febrile neutropenia show similar rates of infection and outcome to younger ones. PMID- 12542478 TI - Incidence of potential glycosylation sites in immunoglobulin variable regions distinguishes between subsets of Burkitt's lymphoma and mucosa-associated lymphoid tissue lymphoma. AB - Recently, a high incidence of novel N-glycosylation sites introduced by somatic mutation was observed in the immunoglobulin variable region genes of follicular lymphoma. As these are positively selected and are uncommon in normal B cells, they may have a role in tumour growth and behaviour. Sites are not characteristic of chronic lymphocytic leukaemia or myeloma, but are detectable in approximately 50% of diffuse large cell lymphomas. Another feature of the variable region genes of certain lymphomas is ongoing somatic mutation. To determine whether glycosylation is associated with this phenomenon, we analysed variable region gene sequences of Burkitt's lymphoma (BL) and mucosa-associated lymphoid tissue (MALT) lymphoma. Novel sites were common in endemic BL (82%) and in 4/5 patients with Iranian BL. However, sporadic BL had a lower incidence (43%). Patients with MALT lymphoma had a low frequency (9%) of novel sites, comparable to normal B cells. These findings distinguish glycosylation sites from ongoing mutation and may reflect different environmental influences on these tumours. PMID- 12542479 TI - A possible role for the loss of CD27-CD70 interaction in myelomagenesis. AB - CD27 is a marker of memory B cells and its interaction with its ligand, CD70, is very important for differentiation into plasma cells. Although CD27 is detected on normal plasma cells, its expression is significantly reduced with the progression of multiple myeloma (MM), including monoclonal gammopathy of undetermined significance (MGUS). CD27+ myeloma cells are thought to represent an early phase of myeloma, as CD27+ plasma cells from MM patients were found to be composed of normal plasma cells (CD19+/CD38++) and myeloma cells (CD19-/CD38++), and monoclonality was detected in the CD27+/CD38++ fraction. Given that the lack of CD27 on plasma cells is related to myelomagenesis and that the pro-apoptotic protein Siva is thought to bind to the cytoplasmic tail of CD27, we analysed alterations of cell growth and genes caused by co-culturing CD27-transfected myeloma cell lines (U266, KMS-5) with CD70-transfected NIH3T3 cells. CD27-CD70 interaction could not induce apoptosis in either type of myeloma transfectant, and binding between Siva and CD27 was not detected. cDNA microarray (human apoptosis CHIP) analysis showed a significant upregulation of expression of the ectodermal neural cortex 1 (ENC1) gene by CD27-CD70 interaction compared with CD27 transfection alone. These findings show that the relationship between the loss of CD27 and oncogenesis of plasma cells is not simple. It remains unclear whether the lack of CD27 leads to evasion of apoptosis. PMID- 12542480 TI - Comparison of five biochemical markers of bone resorption in multiple myeloma: elevated pre-treatment levels of S-ICTP and U-Ntx are predictive for early progression of the bone disease during standard chemotherapy. AB - Increased osteoclastic bone resorption is the major causal factor of bone disease in multiple myeloma. Recently, non-invasive methods have been developed for the estimation of bone resorptive activity. To evaluate the biological sensitivity and clinical usefulness of five biochemical assays for measuring the C-terminal telopeptide of collagen I (ICTP) in serum (beta-Crosslaps ELISA and ICTP radioimmunoassay) and urinary creatinine-adjusted excretions of pyridinoline (PYR), deoxypyridinoline (DPD) and N-terminal telopeptide of collagen I (Ntx), we performed a study of 34 consecutive newly diagnosed myeloma patients. Serum and morning-fasting, second-void urine samples were taken before the start of treatment. In total, 40 age- and sex-adjusted healthy individuals served as controls. Results were expressed as Z-scores. All test variables were highly significantly elevated in the patients (P < 0.001). Serum (S)-ICTP was elevated (Z-score > 2) in most patients (85%) and showed significantly higher Z-score values than the other markers. S-ICTP remained more sensitive than the urinary assays when patients with impaired renal function were excluded from analysis. S ICTP and the urinary metabolites correlated significantly with skeletal morbidity. S-beta-Crosslaps correlated with the bone morbidity only when patients with renal insufficiency were excluded from the analysis. High levels of S-ICTP and urinary (U)-Ntx correlated with an increased risk for early progression of bone lesions during standard melphalan-prednisolone treatment. U-Ntx and S-ICTP are sensitive tools for estimating the increased bone resorption in multiple myeloma and are clinically useful for identifying patients with increased risk of early progression of bone disease. PMID- 12542481 TI - Idiotype-pulsed dendritic cells are able to induce antitumoral cytotoxic CD8 cells in chronic lymphocytic leukaemia. AB - Idiotypic structures of immunoglobulins from malignant B cells constitute tumour specific antigens, though the function of immunoglobulin-specific CD8+ T cells in disease control and rejection remains unclear. We have studied five cases of B chronic lymphocytic leukaemia patients affected with indolent (three patients) or aggressive (two patients) disease. We showed that CD8+ T cells with major histocompatibility complex class I-restricted cytotoxicity against autologous tumour B cells could be generated following repeated stimulations with idiotype pulsed dendritic cells in vitro. CD8+ T-cell lines were able to upregulate CD69 expression and to release interferon (IFN)-gamma upon contact with the autologous B cells, though cytolytic activity was only substantiated for patients with indolent disease. The failure of cytolytic activity in patients with aggressive disease may be explained by a skewed maturation of memory CD8 cells. PMID- 12542482 TI - Novel translocations that disrupt the platelet-derived growth factor receptor beta (PDGFRB) gene in BCR-ABL-negative chronic myeloproliferative disorders. AB - The BCR-ABL-negative chronic myeloproliferative disorders (CMPD) and myelodysplastic/myeloproliferative diseases (MDS/MPD) are a spectrum of related conditions for which the molecular pathogenesis is poorly understood. Translocations that disrupt and constitutively activate the platelet-derived growth factor receptor beta(PDGFRB) gene at chromosome band 5q33 have been described in some patients, the most common being the t(5;12)(q33;p13). An accurate molecular diagnosis of PDGFRB-rearranged patients has become increasingly important since recent data have indicated that they respond very well to imatinib mesylate therapy. In this study, we have tested nine patients with a CMPD or MDS/MPD and a translocation involving 5q31-33 for disruption of PDGFRB by two-colour fluorescence in situ hybridization (FISH) using differentially labelled, closely flanking probes. Normal control interphase cells gave a false positive rate of 3% (signals more than one signal width apart). Six patients showed a pattern of one fused signal (from the normal allele) and one pair of signals separated by more than one signal width in > 85% of interphase cells, indicating that PDGFRB was disrupted. These individuals had a t(1;5)(q21;q33), t(1;5)(q22;q31), t(1;3;5)(p36;p21;q33), t(2;12;5)(q37;q22;q33), t(3;5) (p21;q31) and t(5;14)(q33;q24) respectively. The remaining three patients with a t(1;5)(q21;q31), t(2;5)(p21;q33) and t(5;6)(q33;q24-25) showed a normal pattern of hybridization, with > or = 97% interphase cells with two fusion signals. We conclude that two-colour FISH is useful to determine the presence of a PDGFRB rearrangement, although, as we have shown previously, this technique may not detect subtle complex translocations at this locus. Our data indicate that several PDGFRB partner genes remain to be characterized. PMID- 12542483 TI - Overexpression of complement receptors and related antigens on the surface of bone marrow mast cells in patients with systemic mastocytosis. AB - Depending on their stage of maturation and other factors, mast cell (MC) subsets differ from each other in terms of the expression of complement-associated antigens. This study analysed the expression of various complement-related cell surface antigens (CD11b/CR3, CD11c/CR4, CD35/CR1, CD55/DAF, CD59/MIRL, CD88/C5aR) on bone marrow mast cells (BMMC) in patients suffering from systemic mastocytosis (SM), other haematological diseases and non-haematological disorders (control groups). Expression of complement-associated cell surface antigens was analysed by flow cytometry. There were clear immunophenotypic differences between BMMC obtained from patients with SM and those from the control subjects: the percentage of patients expressing surface CD11c, CD35 and CD88 was significantly higher in patients with SM (76%, 100%, 54%) than in the control subjects (58%, 11%, 18%) (P < 0.05). In addition, the levels of CD11c, CD35 and CD88 expressed per MC (sites per cell) were significantly higher (P < 0.05) in SM than in the control group. Expression of the complement regulatory molecules CD55 and CD59 was detected in BMMC in all patients analysed. However, the levels of CD59 per BMMC were higher in patients with SM as compared with the control subjects, which could help to explain the formation of BMMC aggregates in the former group of individuals. Together, our results showed that BMMC in systemic mastocytosis overexpressed the cell surface membrane receptors involved in binding of complement components and complement-mediated cell activation. Whether this pathological expression of complement receptors is of pathophysiological significance remains to be determined. PMID- 12542484 TI - Early detection of meningeal localization in acute promyelocytic leukaemia patients with high presenting leucocyte count. AB - Extramedullary relapse occurs infrequently in acute promyelocytic leukaemia (APL) but has been increasingly reported after the advent of all-trans retinoic acid (ATRA) treatment, probably as a consequence of improved patient survival. We describe our single centre experience of six APL patients who had disease localization in the central nervous system (CNS). In three patients, clinical symptoms (headache and/or nausea) that presented during follow-up led to the performance of a lumbar puncture and detection of overt CNS infiltration. Two of these patients had simultaneous haematological relapse and one was in molecular remission when CNS leukaemia was documented. One patient with no local symptoms showed CNS infiltration at the time of molecular relapse. Following the introduction of routine lumbar puncture, carried out after front-line induction in all newly diagnosed patients with white blood cell count (WBC) greater than 10 x 109/l, two additional patients in molecular remission with no local symptoms were found to have initial APL localization in the CNS. Presenting features included in 6/6 patients an elevated WBC count (> 10 x 109/l) and a predominance of the PML/RAR bcr3 type (5/6 patients) and of microgranular morphology (5/6 patients). Our findings highlight the importance of carrying out lumbar puncture in APL patients presenting with high-risk features. PMID- 12542485 TI - A novel fusion variant of the MORF and CBP genes detected in therapy-related myelodysplastic syndrome with t(10;16)(q22;p13). AB - We report a case of therapy-related myelodysplastic syndrome (t-MDS) with t(10;16)(q22;p13), in which novel fusion transcripts of the MORF and CBP genes were detected. In one MORF-CBP fusion transcript, exon 15 of the MORF gene was fused in frame with exon 5 of the CBP gene. In a reciprocal CBP-MORF transcript, exon 4 of the CBP gene was fused in frame with exon 16 of the MORF gene. This is the first reported case of t-MDS associated with t(10;16), and provides molecular evidence that the novel MORF-CBP and/or CBP-MORF fusion protein(s) might play an important role in the development of t-MDS. PMID- 12542486 TI - Successful treatment of acute myelomonocytic leukaemia with NUP98-HOXD11 fusion transcripts and monitoring of minimal residual disease. AB - Patients with haematological malignancies involving the NUP98 gene have been reported to have an aggressive clinical course and a poor outcome. We report successful treatment of a 15-year-old Japanese boy with acute myelomonocytic leukaemia having t(2;11)(q31;p15) and a novel fusion transcript, NUP98-HOXD11. He achieved complete remission by combined chemotherapy, and underwent unrelated cord blood transplantation 4 months after diagnosis. He is in complete remission 24 months after diagnosis. Monitoring of minimal residual disease (MRD) showed the absence of fusion transcript 12 months after transplantation. This is the first report of monitoring MRD in a patient with haematological malignancy involving NUP98 fusion transcripts. PMID- 12542487 TI - Risk-adjusted prognostic models for Hodgkin's disease (HD) and grade II non Hodgkin's lymphoma (NHL II): validation on 6728 British National Lymphoma Investigation patients. AB - Using significant factors from multivariate analyses, based on 20 putative markers from a consecutive series of 1198 Sheffield Lymphoma Group patients, risk adjusted prognostic models had been previously derived for Hodgkin's disease (HD) (using age, albumin and lymphocyte count) and non-Hodgkin's lymphoma (NHL) grade II (based on albumin, age, erythrocyte sedimentation rate, lactate dehydrogenase and stage). Data from 6728 patients on the British National Lymphoma Investigation database were used for validation: thus the models were applied to 4411 patients with HD and 2317 patients with NHL grade II. Survival curves derived from these validation groups confirmed our risk models. PMID- 12542488 TI - Non-myeloablative allografting from human leucocyte antigen-identical sibling donors for treatment of acute myeloid leukaemia in first complete remission. AB - Many patients with acute myeloid leukaemia (AML) in first complete remission (CR1) are ineligible for allogeneic transplantation as a result of age or medical problems other than leukaemia. Eighteen patients (median age 59 years, range 36 73 years) with de novo (n = 13) and secondary (n = 5) AML in morphological CR1, who were not candidates for conventional allografting, received non-myeloablative peripheral blood stem cell transplants from human leucocyte antigen identical sibling donors after conditioning with 2 Gy total body irradiation (TBI; n = 10) or 2 Gy TBI and 90 mg/m2 of fludarabine (n = 8). Postgrafting immunosuppression was with cyclosporine and mycophenolate mofetil. Two rejections were observed in patients not given fludarabine and one died with relapse. Overall, 10 patients died between 77 and 841 d, seven from relapse and three from non-relapse mortality (NRM). Day +100 NRM was 0% with a 1-year estimated NRM of 17%[95% confidence interval (CI) 0-35%]. The median follow-up among the eight survivors was 766 d (range, 188-1141 d). Seven of these eight survivors remain in complete remission (CR). One-year estimates of overall and progression-free survivals were 54% (95% CI 31-78%) and 42% (95% CI 19-66%) respectively. While follow-up is short, this analysis demonstrates that the procedure is sufficiently safe to be studied in a wider group of patients. PMID- 12542489 TI - Bone marrow transplantation for beta-thalassaemia major: the UK experience in two paediatric centres. AB - Stem cell transplantation (SCT) remains the only cure for thalassaemia major. Recent advances in medical treatment make it even more important that accurate information is available regarding outcome of SCT in relevant patient populations in order to guide informed decisions regarding the most appropriate treatment for individual thalassaemia patients. We report the results of 55 consecutive first related allogeneic bone marrow transplants (BMT) for children with beta thalassaemia major performed in two UK paediatric centres over 10 years. Between February 1991 and February 2001, 55 children underwent 57 allogeneic BMT. The median age at BMT was 6.4 years and the majority of patients (73%) originated from the Indian subcontinent. Using the Pesaro risk classification, 17 patients were class 1, 27 were class 2 and 11 were class 3. Actuarial overall survival and thalassaemia-free survival at 8 years were 94.5% (95% CI 85.1-98.1) and 81.8% (95% CI 69.7-89.8) respectively. Despite the majority of patients being in class 2 or 3, transplant-related mortality was low (5.4%). The principal complication was graft rejection accompanied by autologous reconstitution that occurred in 13.2% of transplants. Following modification of the conditioning regimen in 1993, the rejection rate fell to 4.6% and remained low. Acute graft-versus-host disease (GVHD) of grade II-IV occurred in 31% and chronic GVHD in 14.5%. These data compare favourably with survival with medical treatment for thalassaemia major and suggest that allogeneic BMT remains an important treatment option for children with beta-thalassaemia major, particularly when compliance with iron chelation is poor. PMID- 12542490 TI - Tandem transplants with different high-dose regimens improve the complete remission rates in multiple myeloma. Results of a Grupo Espanol de Sindromes Linfoproliferativos/Trasplante Autologo de Medula Osea phase II trial. AB - Between 1994 and 1999, 88 multiple myeloma (MM) patients were included in a phase II study to evaluate a tandem autologous stem cell transplantation (ASCT) programme. The first was conditioned with melphalan 200 mg/m2 (MEL200-ASCT1), and the second with cyclophosphamide, etoposide and BCNU (CBV-ASCT2). All patients were in response after MEL200-ASCT1. A control group of MM patients with response to a single ASCT was selected to compare outcomes. After MEL200-ASCT1, 26 patients (30%) achieved complete remission (CR). Of the remaining 48 evaluable patients, 16 (33%) achieved CR with CBV-ASCT2. The final CR rate was 48%. The 5 year survival (OS) was 55%[95% confidence interval (CI) 43-67%] while the event free survival (EFS) was 28% (95% CI 15-39%). CR status after CBV-ASCT2 was the most important prognostic factor for OS and EFS (P = 0.00001), although no differences in outcomes were detected when the patients in CR after MEL200-ASCT1 were compared with those who obtained CR after CBV-ASCT2. Univariate and multivariate analyses showed improved OS and EFS for the tandem series as compared with the control series treated with a single MEL200-ASCT. However, in a stratified comparison by response, there were no prognostic differences between tandem patients and control patients treated with a single ASCT. In summary, our study suggests that the benefit of a second high-dose therapy course depends on its capacity to result in CR for MM patients who have not attained CR after ASCT1. PMID- 12542491 TI - Allogeneic haematopoietic stem cell transplantation for the treatment of adult T cell leukaemia/lymphoma. AB - The feasibility of allogeneic haematopoietic stem-cell transplantation (allo HSCT) in 11 patients with adult T-cell leukaemia/lymphoma (ATL) (6 acute, 4 lymphoma, 1 chronic type) was evaluated. The preparative regimens (9 conventional, 2 reduced-intensity) were tolerable. Five patients developed acute graft-versus-host disease (GVHD), and three, extensive chronic GVHD. All 10 patients who survived > 30 d achieved complete remission. Estimated 1-year overall and disease-free survival rates were 53 +/- 30% and 45 +/- 29% respectively. Four patients remain alive and disease-free at a median follow-up of 25 months. The others died of transplantation-related complications. This pilot study suggests that allo-HSCT in ATL should be evaluated further. PMID- 12542492 TI - Is the factor XIII 34Val/Leu polymorphism a protective factor for cerebrovascular disease? AB - A frequent polymorphism in the factor XIII (FXIII) A-subunit gene, leading to a Val to Leu amino acid exchange at position 34, suggested to affect clot stability, has been associated with a decreased risk for venous thromboembolism and myocardial infarction. Its role in the development of stroke is still under investigation. Ninety-four patients with primary arterial intracerebral haemorrhage (mean age +/- standard deviation: 69 +/- 14 years; 48 men, 46 women), 718 patients with ischaemic stroke (63 +/- 14 years; 395 men, 323 women) and 369 healthy control subjects (59 +/- 14 years; 299 men, 170 women) were analysed for FXIII Val34Leu. No differences in genotype distribution between all three groups were observed. Also, no significant differences in the genotype distribution were found between subgroups of patients stratified according to age, sex, aetiology, history of hypertension, antiplatelet or anticoagulant medication and other vascular risk factors. In contrast to previously reported findings in smaller collectives, our data suggest that an association of the FXIII Val34Leu polymorphism with a decreased risk of ischaemic stroke or an increased risk of intracerebral haemorrhage is highly unlikely. Thus, screening for the FXIII Val34Leu polymorphism will not contribute significantly to the risk prediction of cerebrovascular disease. PMID- 12542493 TI - The effects of chemotherapeutic agents on the regulation of thrombin on cell surfaces. AB - Thromboembolic disorders are common in cancer patients. Two major contributing factors are central venous catheters for drug delivery and the use of l aparaginase, which decreases the plasma antithrombin level, but the causes of the hypercoagulable state in these patients are not fully understood. In this study, the T24/83 cell line was used as a model to investigate the effects of chemotherapeutic agents on cell surface thrombin regulation. Plasma thrombin generation and prothrombin consumption was increased in most of the treated cells, particularly vincristine- and adriamycin-treated cells (P < 0.05), compared with controls. However, no free thrombin generation or prothrombin consumption was observed in factor VII (FVII)-depleted plasma. No significant differences in the levels of thrombin-alpha2-macroglobulin (IIa-alpha2M) and thrombin-anti-thrombin (TAT) were observed between controls and any of the treatments, except for vincristine- and adriamycin-treated cells, which showed a significant difference in TAT production (P < 0.05). Also, there was an upregulation in tissue factor (TF) mRNA expression in etoposide-, methotrexate- and vincristine-treated monolayers compared with controls, as well as an upregulation in TF protein production in vincristine-treated cells. The data suggests that thrombin generation occurs via the extrinsic (TF-dependent) coagulation pathway on cell surfaces and that some chemotherapeutic agents are able to upregulate TF mRNA and protein expression in T24/83 cells. PMID- 12542494 TI - Anti-thymocyte globulin treatment of marrow aplasia associated with paroxysmal nocturnal haemoglobinuria (PNH) resulted in haematological improvement due to an expansion of the PNH clone. AB - A patient with aplastic anaemia developed paroxysmal nocturnal haemoglobinuria (PNH) 4 years after diagnosis, with an ensuing haematopoietic improvement. The PNH clone subsequently declined, leading to pancytopenia again. Anti-thymocyte globulin had to be administered 14 years later, which resulted in haematopoietic improvement once more. Flow cytometric analysis showed that this was attributable to expansion of the PNH clone, owing probably to alleviation of its suppression by immune-mediated mechanisms. PIG-A gene analysis showed that the same PNH clone had waned and waxed in the clinical course. Our results suggest that the PNH clone might rarely be an immune target as well. PMID- 12542495 TI - Granulocyte colony-stimulating factor administration upregulates telomerase activity in CD34+ haematopoietic cells and may prevent telomere attrition after chemotherapy. AB - Hematopoietic reconstitution could be associated with premature ageing of the transplanted cells and a high frequency of myelodysplastic syndrome and secondary leukaemia. Telomere length decreases with cell divisions and age, and at a crucial length it is associated with chromosomal instability and cell senescence. Telomerase is a reverse transcriptase enzyme that adds nucleotides to chromosomal ends. Most somatic cells lack telomerase activity yet haematopoietic stem cells retain low levels of telomerase. Some studies have found that chemotherapy and stem cell transplantation lead to the accelerated shortening of telomere length. As granulocyte colony-stimulating factor (G-CSF) is routinely used in the mobilization of stem cells for transplantation, we evaluated its effects on telomerase activity and regulation, and on telomere dynamics, in normal donors and selected lymphoma patients. Administration of G-CSF increased telomerase activity in CD34+ haematopoietic cells compared with controls. In marrow-derived CD34+ cells, telomerase activity increased sevenfold, compared with a 14-fold increase in peripheral-blood-mobilized CD34+ cells. A parallel increase in the expression of human telomerase enzyme reverse transcriptase RNA and protein kinase C alpha occurred. In addition, G-CSF administration to five lymphoma patients after consecutive courses of CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone) chemotherapy, resulted in telomere length preservation or elongation, as opposed to marked attrition in patients who did not receive growth factors. We conclude that the in vivo administration of G-CSF prevents or attenuates telomere attrition associated with chemotherapy administration. This attenuation may contribute to the preservation of telomere integrity inG-CSF primed transplanted stem cells. PMID- 12542496 TI - HIV-1 matrix protein p17 enhances the proliferative activity of natural killer cells and increases their ability to secrete proinflammatory cytokines. AB - We investigated the effects of human immunodeficiency type-1 virus (HIV-1) matrix protein p17 on freshly isolated and purified human natural killer (NK) cells. HIV 1 p17 increased the cytokines interleukin (IL) 2, IL-12 and IL-15, and induced natural killer cell proliferation, but not cytotoxicity. This effect was specific because it was abrogated by anti-p17 monoclonal antibody. Moreover, HIV-1 p17 enhanced the cytokine-induced production of tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma by NK cells. IL-4 downregulated IFN-gamma and TNF alpha secretion in IL-2- and IL-15-treated NK cells. HIV-1 p17 restored the ability of NK cells to produce both cytokines when added to the cultures simultaneously with IL-4. The property of p17 to increase the production of TNF alpha and IFN-gamma might be a mechanism used by HIV-1 to modulate the immune system to support its replication and spreading. PMID- 12542497 TI - Regulated expression of MUC1 epithelial antigen in erythropoiesis. AB - MUC1 is a large surface glycoprotein expressed by epithelial cells, which is overexpressed and aberrantly glycosylated in carcinomas. MUC1 is involved in epithelial cell interactions and appears to function as a signal-transducing molecule. The finding that MUC1 can also be expressed in the haematopoietic lineages prompted us to further investigate the possible function(s) of this molecule in haematopoietic cells. In bone marrow differentiating cells, MUC1 was strongly and selectively expressed during erythropoiesis; it was also weakly expressed during megakaryocytopoiesis and granulomonocytopoiesis; however, no correlation between MUC1 and differentiation marker expression was observed in these lineages. In vitro CD34+ cells, induced towards erythroid differentiation, acquired MUC1 transiently, while expressing increasing levels of the lineage marker glycophorin A. MUC1 was absent in the circulating erythrocytes. During erythropoiesis, MUC1 expression was transcriptionally regulated and the molecule underwent phosphorylation. To investigate the possible role of MUC1 during erythropoiesis, we studied the ability of MUC1 to act as ligand for cell-cell interaction. The sialylated MUC1 glycoforms selectively expressed on erythroid cells were able to bind the macrophage-restricted molecule sialoadhesin. These results suggest that MUC1 can function as a cross-talk molecule between the erythroblasts and the surrounding cells during erythropoiesis. PMID- 12542498 TI - Impact of disordered puberty on bone density in beta-thalassaemia major. AB - Reduction of bone density and its associated morbidity is recognized in young adults with beta-thalassaemia major, but the aetiology is not clear. This study used dual X-ray absorptiometry (DXA) to look at bone mineral apparent density (BMAD) in children and young adults with thalassaemia in a predominantly Asian population, in the context of sexual maturation. Fifty-five patients were scanned (mean age 13.8 years, range 5.9-37.5) and BMAD z-scores were calculated using normal data from locally recruited control subjects. Eighteen patients had undergone bone marrow transplantation (BMT) and the remainder were on a transfusion/chelation regimen. BMAD z-scores ranged from -3.3-1.6 with a mean of 0.92. No difference in BMAD was found between those patients treated conventionally and those who had undergone BMT. When comparing mean BMAD z-score according to sexual maturation, there was a highly significant difference (P < 0.0001) between those whose pubertal maturation was age appropriate (mean z-score -0.22), when compared with those who had disordered puberty (mean z-score -1.82). We have shown that failure to progress normally through puberty is highly significant in the failure of adequate bone mineralization and achievement of peak bone mass in thalassaemic patients. The management of these patients should therefore be pro-active to anticipate problems and facilitate normal sexual maturation. PMID- 12542499 TI - Circadian variations of transferrin saturation levels in iron-overloaded patients: implications for the screening of C282Y-linked haemochromatosis. AB - The phenotypic screening for genetic haemochromatosis (GH) relies upon the determination of transferrin saturation (TS). In large-scale screening programs, the time of blood sampling can be uneasy to control. We studied the circadian variations of TS at 08.00 hours, 12.00 hours, 18.00 hours and 00.00 hours in 46 C282Y homozygous patients (GH) and 47 non-GH patients (NH), to determine whether the time of blood sampling influenced the results of screening. In both groups, there were significant circadian variations in TS, with the highest values at 08.00 hours and the lowest at 00.00 hours. For any given time-point, TS was significantly higher in the GH group when compared with the NH group (P < 0.0001). For both groups, there was a significant decrease in TS between 08.00 hours and 00.00 hours (P < 0.0001) but this decrease was not as significant in GH when compared with NH patients (interaction P < 0.0073). Receiver operating characteristics (ROC) curves generated for TS at 08.00 hours, 12.00 hours, 18.00 hours and 00.00 hours, presented the same efficiency of diagnosis of GH, with TS threshold varying between 64% at 08.00 hours and 36% at 00.00 hours. In conclusion, for screening studies of C282Y homozygosity, determination of transferrin saturation may be performed at any time during the day. PMID- 12542500 TI - A novel 7.9 kb deletion causing alpha+-thalassaemia in two independent families of Indian origin. AB - We describe the characterization of a novel 7.9 kb deletion that eliminated one of the duplicated alpha-globin genes, causing an alpha+-thalassaemia phenotype in two independent carriers of Suriname-Indian origin. The molecular characterization of the deletion breakpoint fragment revealed neither involvement of Alu repeat sequences nor the presence of homologous regions prone to recombination, suggesting a non-homologous recombination event. This alpha+ thalassaemia deletion was found to give rise to an atypical haemoglobin H (HbH) disease characterized by a non-transfusion-dependent moderate microcytic hypochromic anaemia in combination with a poly adenylation signal mutation of the alpha-globin gene (alpha2 AATAAA --> AATA-- --). PMID- 12542501 TI - Long haul flights and deep vein thrombosis: who is at risk? PMID- 12542502 TI - Severe aplastic anaemia in a child with brachydactyly type C. PMID- 12542504 TI - Modified chop-chemotherapy plus rituximab for diffuse large b-cell lymphoma complicating ataxia-telangiectasia. PMID- 12542506 TI - The influence of fatigue on health-related quality of life in patients with Parkinson's disease. AB - OBJECTIVE: To examine the correlation between fatigue and health-related quality of life (HRQL) in patients with Parkinson's disease (PD). PATIENTS AND METHODS: Sixty-six patients with idiopathic PD. The patients did not have a depressive mood disorder or cognitive impairment. Fatigue was measured by the Fatigue Severity Scale (FSS). HRQL was measured by the Parkinson's Disease Questionnaire (PDQ-39) and the Short-Form 36 (SF-36). RESULTS: Thirty-three (50%) of the patients had significant fatigue. Patients with fatigue had a more advanced disease than those without fatigue, measured by the UPDRS scale, including a higher Hoehn and Yahr stage and lower Schwab and England score. Patients with fatigue reported more distress in the dimensions of emotional well-being and mobility (PDQ-39) and also had a significantly higher PDQ summary index. On the SF-36 patients with fatigue reported more problems in the areas of physical functioning, role limitation (physical), social functioning and vitality. Correlations between the FSS and the HRQL scales were highest for the summary index of PDQ-39 and in the dimensions of ADL, mobility and emotional well-being (PDQ-39) and physical functioning, role limitation (physical), social functioning, general health and vitality (SF-36). CONCLUSIONS: PD has a substantial negative impact on HRQL. We found a strong correlation between fatigue and high distress scores on HRQL scales in a population of patients with PD who were not depressed or demented. The diversity of symptoms and high prevalence of non-motor features, including fatigue, is important to take into account in our efforts to optimize treatment and care for this patient group. PMID- 12542507 TI - Clinical benefits of interferon beta-1a in relapsing-remitting MS: a phase IV study. AB - OBJECTIVE: To evaluate the efficacy and safety of IFNbeta-1a (Avonex, Biogen, Inc., Cambridge, MA, USA) in patients with relapsing-remitting multiple sclerosis (MS). METHODS: In this multicenter, open-label, prospective clinical trial, 96 patients with relapsing-remitting MS received IFNbeta-1a 30 mcg intramuscularly once weekly for 2 years. Outcome variables included: change from baseline in mean number of exacerbations, proportion of exacerbation-free patients, and mean Expanded Disability Status Scale (EDSS) scores at Years 1 and 2. RESULTS: IFNbeta 1a significantly (P < 0.0001) reduced exacerbation rate at Years 1 and 2 of treatment. The percentage of exacerbation-free patients was 53% during Year 1 and 33% during Year 2. Mean EDSS scores were 2.96 +/- 1.26 at baseline, 2.89 +/- 1.42 at Year 1, and 3.00 +/- 1.62 at Year 2 (P = 0.116). EDSS scores improved in 35.4%, remained stable in 28.1%, and worsened in 36.5% of patients. IFNbeta-1a treatment was well tolerated. CONCLUSION: This study confirms and extends the beneficial clinical profile for IFNbeta-1a in relapsing MS. PMID- 12542508 TI - Surgical prognosis in hindbrain related syringomyelia. AB - OBJECTIVES: The recommended operative treatment against hindbrain related syringomyelia is suboccipital decompression. The aim of the study was to define prognostic factors for surgical outcome in this disease. MATERIALS AND METHODS: Between 1990 and 1997, 31 patients with hindbrain related syringomyelia were treated. All patients underwent craniovertebral decompression by suboccipital craniectomy and laminectomy of C1 or C1 and C2, respectively. Additionally, in nearly half of the cases, the tonsils were treated by bipolar coagulation. Mean post-operative observation period was 35 months including clinical and radiological (MRI) examination. RESULTS: Neither there was correlation between clinical outcome and age nor correlation between clinical outcome and duration of preoperative symptoms. There was good correlation between clinical outcome and result of post-operative MRI: 63% of patients with a sufficient post-operative MRI demonstrated a significant clinical improvement, whereby only 17% of patients with insufficient MRI did so (P < 0.05). None of the patients with coagulation of the tonsils showed clinical improvement. In contrast, 77% of patients without tonsillar manipulation demonstrated clinical improvement (P < 0.01). Additionally, 88% of the patients with dorsal-tenting duraplasty experienced neurological improvement (P < 0.01), too. CONCLUSIONS: The results give evidence for the prognostic importance of creating an adequate artificial cisterna magna. Dorsal-tenting of the duraplasty is an advantageous means against scarring and adhesions, whereby intradural applications such as extensive bipolar coagulation of the cerebellar tonsils should be avoided. PMID- 12542509 TI - Developmental dyslexics show altered allocation of attention in visual classification tasks. AB - OBJECTIVES: Event-related brain potentials (ERPs) were recorded to investigate allocation of attention in adult developmental dyslexics. SUBJECTS AND METHODS: Twelve adult developmental dyslexics and 12 matched normal controls performed three visual choice reaction tasks. In the passive oddball condition, subjects watched two different simple visual stimuli presented with 87.5 and 12.5% probability. In the active oddball condition, participants responded to the low probability target stimulus. In the active 50/50-condition, both stimuli were presented with 50% probability and a response was required to the target stimulus only. RESULTS: No group differences emerged for performance, P300 latency or laterality and for N200 amplitude, latency or laterality. An enhancement of P300 amplitude with a frontal distribution was found for NoGo (standard)-stimuli in both active conditions for the dyslexic sample. CONCLUSION: Results are discussed in the context of deviances in allocation of attentional resources in dyslexic readers. PMID- 12542510 TI - Gap junction protein beta 1 (GJB1) mutations and central nervous system symptoms in X-linked Charcot-Marie-Tooth disease. AB - OBJECTIVES: To clarify the clinical variability, including central nervous system (CNS) involvement, in X-linked Charcot-Marie-Tooth disease (CMTX) patients. MATERIAL AND METHODS: We clinically, pathologically and genetically studied six CMTX patients with distinct symptoms and four different GJB1 mutations. RESULTS: One patient with Val63Ile had deafness, low intelligence, saccadic eye movement, upper extremity distal dominant muscle weakness and normal sensation. Another patient with Glu186Lys had severe sensorineural deafness at the age of 6 years, but did not develop muscle weakness until the age of 20 years. Two patients with Arg22Gln had typical CMT1A-like clinical features, no CNS symptoms and obvious onion bulb formations. Two siblings with deletion of the entire GJB1 gene had mild to moderate lower extremity muscle weakness and sensory disturbance without CNS involvement. CONCLUSION: These findings suggest that some gain of function mutations of GJB1 may be related to CNS symptoms because the patients with GJB1 deletion only had peripheral neuropathy, although other unknown associated factors may contribute to their clinical phenotypes. PMID- 12542511 TI - Frequency of spinocerebellar ataxia mutations in the Kinki district of Japan. AB - OBJECTIVES: To determine the frequencies of spinocerebellar ataxias (SCAs) in the Kinki district, the western part of the main island of Japan. MATERIAL AND METHODS: One hundred and forty-three families with dominantly inherited ataxia and 220 patients with apparently sporadic cerebellar ataxia were examined for the SCA1, SCA2, SCA3/Machado-Joseph disease (MJD), SCA6, SCA7, SCA8, SCA12 and dentatorubral-pallidoluysian atrophy (DRPLA) mutations. RESULTS: Among the dominant families, SCA1 accounted for 3%, SCA2 for 4%, SCA3/MJD for 24%, SCA6 for 31% and DRPLA for 12%. Neither SCA7 nor SCA12 mutations were detected. Among the apparently sporadic patients, 15% were found to have expanded triplet repeats. Of these, the SCA6 mutation was most frequently detected. CONCLUSION: SCA6 is the most common SCA in the Kinki district of Japan. Comparison of our results with those from other regions of Japan and different countries shows geographic and ethnic variation in the frequency of SCAs. PMID- 12542512 TI - R-R interval variation and the sympathetic skin response in the assessment of the autonomic nervous system in leprosy patients. AB - OBJECTIVES: The aim of this study was to evaluate possible autonomic nervous system (ANS) dysfunction in leprosy patients with the sympathetic skin response (SSR) and the heart rate (R-R) interval variation (RRIV) measurements which are easy and reliable methods for evaluation of autonomic functions. MATERIAL AND METHODS: We studied 37 lepromatous leprosy patients (mean age: 38 +/- 17 years, range 23-62 years, 20 females and 17 males) and 35 age-matched healthy subjects (mean age: 34.19 +/- 12.74 years, range 24-48 years, 20 females and 15 males). Non-invasive bedside tests (orthostatic test, Valsalva ratio), R-R interval variation (RRIV) during at rest and deep breathing, the SSR latency and amplitude from both palms, and nerve conduction parameters were studied in all the subjects. RESULTS: The mean values of RRIV in leprosy patients during at rest [mean RRIV in patients, 17.42 +/- 8.64% vs controls, 22.71 +/- 3.77% (P < 0.05)] and during deep breathing [mean RRIV in patients, 21.64 +/- 9.08% vs controls, 30.70 +/- 5.99% (P < 0.005)] was significantly lower compared with the controls. The mean latency of SSR in leprosy patients [mean SSR latency in patients, 1.72 +/- 1.13 ms vs controls, 1.30 +/- 0.41 ms (P < 0.05)] was significantly prolonged compared with the controls. The mean amplitude of SSR in leprosy patients [mean SSR amplitude in patients, 0.54 +/- 0.57 microV vs controls, 1.02 +/- 0.56 microV (P > 0.05)] was smaller compared with the controls, but this difference was not significant. The mean Valsalva ratio in leprosy patients [mean in patients, 1.11 +/- 0.13 vs controls, 1.16 +/- 0.07 (P > 0.05)] was smaller compared with the controls, but not statistically significant. The mean difference of systolic and diastolic blood pressure between supine rest and during standing in leprosy patients were higher compared with the controls [mean systolic pressure in patients, 7 +/- 6 mmHg vs controls, 6 +/- 8 mmHg (P > 0.05) and mean diastolic pressure in patients, 3 +/- 3 mmHg vs controls, 3 +/- 2 mmHg (P > 0.05)], but they did not reach statistical significance. Furthermore, lower RRIV and the prolonged SSR latencies in leprosy patients were closely correlated to some parameters of sensorimotor nerve conduction and each other [median nerve distal latency and RRIV, r = -0.67 (P < 0.05), ulnar nerve distal latency and RRIV, r = 0.59 (P < 0.05), RRIV and SSR latency, r = -0.33 (P < 0.02)]. These data indicate that leprosy patients have the functional abnormalities of ANS. CONCLUSION: We conclude that combined use of these two tests, both of which can be easily and rapidly performed in the electromyogram (EMG) laboratory using standard equipment, allows separate testing of parasympathetic and sympathetic function, and are very sensitive methods in assessing of ANS function in peripheral neuropathy in leprosy patients. PMID- 12542513 TI - Patterns of mortality in patients with motor neurone disease. AB - OBJECTIVE: Motor neurone disease (MND) is a rapidly fatal condition with survival of less than 4 years. Patients can deteriorate quickly in the preterminal stages resulting in inappropriate resuscitation or admission to intensive care units (ICU) or accident and emergency (A & E). MATERIAL AND METHODS: We looked at patterns of mortality with emphasis on the place of death. A retrospective study was performed of all patients attending an MND clinic, who had died within a 10 year period. RESULTS: Of 179 patients (63 female), 81 patients (45%) died at home, in a hospice or in a nursing home. Sixty-five patients (36%) died in hospital (11 in ICU or A & E). Nine of the latter were previously known to have MND and six admissions were probably avoidable. Most ward patients died of respiratory causes and were treated conservatively. CONCLUSION: The proportion of patients dying in A & E or ICU was small but could have been reduced further. A number of those who died on the wards could probably have been managed conservatively at home. Older patients and those with bulbar disease had a poorer prognosis. PMID- 12542514 TI - Superficial siderosis of the central nervous system: pathogenetic heterogeneity and therapeutic approaches. AB - OBJECTIVE: Superficial siderosis of the central nervous system (CNS) is a rare chronic progressive disorder caused by chronic subarachnoid hemorrhage. We present four patients with superficial siderosis of the CNS to describe the characteristic symptoms, and to discuss the pathogenetic heterogeneity and possible new therapeutic approaches. RESULTS: The causes of chronic subarachnoid bleeding in superficial siderosis were different. In two patients surgical treatment of ependymoma or cerebral cavernomas were the underlying diseases. No cause was detected in one patient. For the first time, we present one patient with vasculitis of the central nervous system associated with systemic hemochromatosis in superficial siderosis. Therapeutic approaches included exstirpation of cavernomas as the source of chronic bleeding in one patient, immunosuppressive therapy and venupunctures in the patient with vasculitis and hemochromatosis, and symptomatic treatment with chelating agents and antioxidants. The patients remained clinically stable for the follow-up period of up to 2 years. CONCLUSIONS: Our cases underline the pathogenetic heterogeneity of superficial siderosis and favor the early diagnosis for prompt initiation of therapy. Besides treatment of the underlying condition, antioxidants and radical scavengers may be effective in halting the progression of the disease. PMID- 12542515 TI - A case of inflammatory demyelinating polyradiculoneuropathy associated with T cell lymphoma. AB - Malignant lymphoma may present prominent peripheral nervous system disorders with variable etiologies. We describe a patient who presented with chronic relapsing polyradiculoneuropathy accompanied by right facial nerve palsy. Gadolinium enhancement of the right facial nerve and cervical spinal roots was noted on magnetic resonance imaging (MRI). Sural nerve biopsy specimens showed mononuclear cell infiltration around the vessels in the epineurium. Histopathological and immunohistochemical investigations of sural nerve specimens revealed perivascular infiltration of lymphocytes with T-cell dominancy. No apparent direct invasion of lymphoma cells was seen. The results of nerve conduction studies, sural nerve biopsy and cerebrospinal fluid examination were suggestive of immune-mediated inflammatory demyelinating neuropathy. The chronic and relapsing fashion and unique radiological findings in our patient expand on the previously reported features of peripheral neuropathy associated with peripheral T-cell lymphoma. PMID- 12542516 TI - Endoscopic ventriculo-cystomy for non-communicating hydrocephalus secondary to quadrigeminal cistern arachnoid cyst. AB - INTRODUCTION: Intracranial arachnoid cysts are developmental, anomalous collections of cerebrospinal fluid (CSF), and occasionally cause symptoms when large enough to obliterate the CSF outflow pathway and give rise to non communicating hydrocephalus. The treatment of choice for symptomatic arachnoid cysts has been surgical excision or fenestration of the cyst, but less invasive endoscopic fenestration has been attempted with favorable preliminary results. CASE REPORT: We report a case of non-communicating hydrocephalus caused by a quadrigeminal cistern arachnoid cyst in a 35-year-old woman who presented with worsening headaches. She was successfully treated by endoscopic third ventriculostomy and fenestration of the cyst (ventriculo-cistomy). DISCUSSION: Endoscopic fenestration of symptomatic arachnoid cysts can be as effective as open surgery in terms of short-term efficacy, and it is certainly less invasive than open surgery. However, its long-term outcome remains unknown, and these rare cases require careful clinical and radiological follow-up. PMID- 12542517 TI - Ross syndrome--a case documentation. PMID- 12542518 TI - Juvenile asymmetric segmental spinal muscular atrophy (Hirayama's disease). PMID- 12542519 TI - Differentiation and apoptosis in human immortalized sebocytes. AB - Increased cell volume, accumulation of lipid droplets in the cytoplasm, and nuclear degeneration are phenomena indicating terminal differentiation of human sebocytes followed by holocrine secretion and cell death. The molecular pathways of natural and induced sebocyte elimination are still unknown, however. In this study, SZ95 sebocytes were found to exhibit DNA fragmentation after a 6 h culture followed by increased lactate dehydrogenase release after 24 h, indicating cell damage. With the help of morphologic studies and using Oil Red detection of cellular lipids, cell enlargement, accumulation of lipid droplets in the cytoplasm, and nuclear fragmentation could be observed under treatment with arachidonic acid. Staurosporine, a potent inhibitor of phospholipid Ca2+ dependent protein kinase, increased externalized phosphatidylserine levels on SZ95 sebocytes, detected by annexin V/propidium iodide flow cytometry, as early as after 1 h, whereas dose-dependent reduction of bcl-2 mRNA and protein expression, enhanced DNA fragmentation, and increased caspase 3 levels, detected by caspase 3 inhibitor/propidium iodide flow cytometry, were found after 6 h of treatment. SZ95 sebocyte death was detected as early as after 6 h of SZ95 sebocyte treatment with high staurosporine concentrations (10(-6)-10(-5) M). 5Alpha-dihydrotestosterone (10(-8)-10(-5) M) did not affect externalized phosphatidylserine levels and DNA fragmentation in SZ95 sebocytes but slightly decreased lactate dehydrogenase cell release. Neither acitretin nor 13-cis retinoic acid (10(-8)-10(-5) M) affected externalized phosphatidylserine levels, DNA fragmentation, and lactate dehydrogenase cell release, despite the increased caspase 3 levels under treatment with 13-cis retinoic acid. The combined staurosporine and 13-cis retinoic acid treatment enhanced DNA fragmentation in SZ95 sebocytes to the same magnitude as in cells only treated with staurosporine. In conclusion, SZ95 sebocytes in vitro undergo apoptosis, which can be enhanced by the terminal differentiation inductor arachidonic acid or by staurosporine and leads to cell death. 5Alpha-dihydrotestosterone inhibits SZ95 sebocyte death without involving apoptotic pathways, and retinoids did not affect the programmed death of human sebocytes. The latter result fits well with the currently reported inability of normal skin cells to undergo apoptosis after treatment with retinoids, in contrast to their malignant counterparts. PMID- 12542520 TI - Fibroblast activation protein: differential expression and serine protease activity in reactive stromal fibroblasts of melanocytic skin tumors. AB - Growth and metastasis of solid neoplasms require the recruitment of a supporting tumor stroma. A highly consistent trait of tumor stromal fibroblasts in most epithelial cancers is the induction of fibroblast activation protein (FAP), a member of the serine protease family. Recently it was demonstrated that FAP has both dipeptidyl peptidase and collagenolytic activity capable of degrading gelatin and type I collagen. In this study, we describe the expression and enzyme activity of FAP in benign and malignant melanocytic skin tumors. FAP-positive fibroblasts were detected immunohistochemically in the reactive stroma of all melanocytic nevi tested. In primary and metastatic melanomas an upregulation of FAP expression in the reactive mesenchyme could be observed. Whereas 30% of the nevi revealed additional FAP expression on subsets of melanocytic cells, melanoma cells from primary and metastatic melanomas were FAP negative. This may indicate a possible role for FAP in the control of tumor cell growth and proliferation during melanoma carcinogenesis. Consistent with this in vivo expression pattern FAP enzyme activity could be detected by a specific immunocapture assay in extracts of melanocytic nevi and melanoma metastases, whereas no significant activity was detectable in normal adult skin. Strong protein expression of FAP was observed in patterned structures restricted to a subset of the melanoma metastases. Our findings that these FAP-positive structures showed no overlap with endothelial cell surface markers, nor with various melanoma antigens, suggest that FAP is a marker for specific stromal-cell-derived patterns in cutaneous melanoma metastases. PMID- 12542521 TI - Plectin-isoform-specific rescue of hemidesmosomal defects in plectin (-/-) keratinocytes. AB - The various plectin isoforms are among the major crosslinking elements of the cytoskeleton. The importance of plectin in epithelia is convincingly supported by the severe skin blistering observed in plectin-deficient humans and mice. Here, we identified plectin 1a (> 500 kDa), a full length plectin variant containing the sequence encoded by the alternative first exon 1a, as the isoform most prominently expressed in human and mouse keratinocytes. In skin sections and cultured keratinocytes, plectin 1a was shown to colocalize with hemidesmosomal structures. In contrast, a second isoform expressed in epithelia, plectin 1c, differing from 1a merely by a short N-terminal sequence, colocalized with microtubules. Expression of plectin 1a, but not of its N-terminal fragment alone, or of a third alternative full length isoform (plectin 1), restored the reduced number of hemidesmosome-like stable anchoring contacts in cultured plectin-null keratinocytes. Our results show for the first time that different isoforms of a cytolinker protein expressed in one cell type perform distinct functions. Moreover, the identification of plectin 1a as the isoform defects in which cause skin blistering in plectin-related genetic diseases, such as epidermolysis bullosa simplex MD and epidermolysis bullosa simplex Ogna, could have implications for the future development of clinical therapies for patients. PMID- 12542522 TI - Light-induced resistance of the keratin network to the filament-disrupting tyrosine phosphatase inhibitor orthovanadate. AB - Epidermal keratinocytes respond to low-dose light irradiation by inducing signaling cascades that lead to long-term effects on gene transcription thereby protecting cells against damage. In contrast, little is known about immediate light-induced alterations of structural proteins. We have made the intriguing observation that light produces fundamental changes in the properties of the keratin filament system of cultured epidermoid A-431 cells. A short light exposure (1-10 min) causes the keratin cytoskeleton to become immediately resistant to the tyrosine phosphatase inhibitor orthovanadate, which otherwise disrupts the keratin filament network completely in just a few minutes. This protective effect is inducible throughout the entire visible spectrum and is elicited by normal room light (<200 Lux). Exposure of cells to monochromatic light of various wavelengths is therefore equally effective. In addition, the acquisition of orthovanadate resistance has been directly monitored in living cells; a partially disrupted keratin cytoskeleton recovers to a completely filamentous network in half an hour. Finally, the protective light effect is largely reversed in 2 h and can be mimicked by preincubation with the p38 kinase inhibitor SB203580. In contrast, the mitogen-activated protein kinase inhibitor PD98059 and epidermal growth factor inhibit orthovanadate action to a lesser extent. Taken together, these observations suggest a stabilizing function of light on the keratin filament network; this may be of relevance to the treatment of skin diseases with reduced keratin stability. PMID- 12542523 TI - The prevalence and clinical significance of anti-U1 RNA antibodies in patients with systemic sclerosis. AB - We studied the prevalence and clinical significance of anti-U1 RNA antibodies in patients with systemic sclerosis. The presence of anti-U1 RNA antibodies was determined using immunoprecipitation in systemic sclerosis patients with anti-U1 RNP antibodies (n=36), antitopoisomerase I antibodies (n=20), or anticentromere antibodies (n=20), mixed connective tissue disease patients (n=23), systemic lupus erythematosus patients with anti-U1 RNP antibodies (n=26), and normal controls (n=20). Moreover, antigen specificities for anti-U1 RNP antibodies were examined in patients with systemic sclerosis by immunoblotting and enzyme-linked immunosorbent assay. Anti-U1 RNA antibodies was detected in 22 of 36 systemic sclerosis patients (61%) with anti-U1 RNP antibodies, 14 of 23 patients (61%) with mixed connective tissue disease, and eight of 26 systemic lupus erythematosus patients (31%) with anti-U1 RNP antibodies. Anti-U1 RNA antibodies were not detected in other groups. As for systemic sclerosis patients, the frequencies of pulmonary fibrosis and reduced percentage diffusion capacity for carbon monoxide were significantly greater in patients with anti-U1 RNA antibodies than in those without (76%vs 18%, p<0.005; 82%vs 27%, p<0.005, respectively). Moreover, patients with anti-U1 RNA antibodies had significantly lower percentage diffusion capacity for carbon monoxide and percentage vital capacity values than those without (51.9+/-16.8 vs 79.4+/-16.4, p<0.01; 83.8+/ 21.4 vs 101.4+/-12.9, p<0.05, respectively). Regarding the antigen specificities of anti-U1 RNP antibodies in systemic sclerosis patients, the frequency of anti 70 kDa antibodies determined by immunoblotting was significantly higher in patients with anti-U1 RNA antibodies than in those without (77%vs 43%, p<0.05). This finding was also confirmed by enzyme-linked immunosorbent assay for anti-70 kDa antibodies (86%vs 43%, p<0.05). These results indicate that anti-U1 RNA antibodies may be a serologic indicator for pulmonary fibrosis in systemic sclerosis patients with anti-U1 RNP antibodies. PMID- 12542524 TI - Risk of malignancies in psoriasis patients treated with cyclosporine: a 5 y cohort study. AB - This prospective long-term cohort study investigated the incidence of malignancies in severe psoriasis patients treated with cyclosporine. A total of 1252 patients were followed prospectively for up to 5 y. Malignancies were recorded prospectively. Incidence rates for malignancies were compared with the general population using standardized incidence ratios. The effect of duration of exposure to cyclosporine and to previously administered anti-psoriatic treatments on the incidence of malignancies was investigated using Poisson regression models. The mean age of patients was 43 y and on average, patients received cyclosporine for 1.9 y. Malignancies were diagnosed in 47 patients (3.8%), 49% of them had skin malignancies. The standardized incidence ratio in the study cohort was 2.1 as compared with the general population. The higher incidence of malignancies was attributed to a 6-fold higher incidence of skin malignancies, most of which were squamous cell carcinoma. The incidence of nonskin malignancy overall was not significantly higher in this study than in the general population. Duration of exposure to cyclosporine, exposure to psoralen and ultraviolet A, exposure to methotrexate, and exposure to immunosuppressants showed a significant effect on the incidence of nonmelanoma skin malignancies. In conclusion, treatment of psoriasis with cyclosporine is associated with an increased risk of nonmelanoma skin cancer. Patients treated for more than 2 y with cyclosporine were shown to have a higher risk. In addition, exposure to psoralen and ultraviolet A and to other immunosuppressants was shown to contribute to the overall risk. PMID- 12542525 TI - Familial and attributable risks in cutaneous melanoma: effects of proband and age. AB - We studied familial risks in invasive and in situ cutaneous melanoma by comparing the occurrence of melanoma, or discordant cancer, between parents and offspring, and between siblings, based on the Swedish Family Cancer Database of over 10 million individuals. Offspring were 0-66 y of age. Cancers were obtained from the Swedish Cancer Registry from 1961 to 1998. The study was based on 24,818 invasive and 5510 in situ cases of melanoma. Standardized incidence ratios were calculated for familial risk. The standardized incidence ratios for offspring was 2.40 (95% confidence intervals: 2.10-2.72) when only the parent had melanoma and it was 2.98 (95% confidence intervals: 2.54-3.47) when only a sibling was affected; when both a parent and a sibling were affected the standardized incidence ratios was 8.92 (95% confidence intervals: 4.25-15.31). The respective population attributable risks were 1.38, 1.20, and 0.10%. The familial risk showed a clear age dependence and somewhat higher risk in in situ melanoma than in the invasive counterpart. The highest standardized incidence ratio of 61.78 (5.82-227.19) was noted for offspring whose parent had multiple melanomas. Superficially spreading melanoma showed the highest familial risk both among invasive and in situ tumors. Melanoma associated with breast, nervous system, and skin cancers, and in situ melanoma possibly also with connective tissue and thyroid tumors and multiple myeloma. PMID- 12542526 TI - Self-healing collodion baby: a dynamic phenotype explained by a particular transglutaminase-1 mutation. AB - Spontaneous healing with no or only very mild ichthyosis distinguishes the "self healing collodion baby" from other congenital ichthyoses. In two self-healing collodion baby siblings with markedly diminished epidermal transglutaminase 1 activity we found the compound heterozygous transglutaminase 1 mutations G278R and D490G. Molecular modeling and biochemical assays of mutant proteins under elevated hydrostatic pressure suggest significantly reduced activity in G278R and a chelation of water molecules in D490G that locks the mutated enzyme in an inactive trans conformation in utero. After birth these water molecules are removed and the enzyme is predicted to isomerize back to a partially active cis form, explaining the dramatic improvement of this skin condition. PMID- 12542527 TI - Acrokeratosis verruciformis of Hopf is caused by mutation in ATP2A2: evidence that it is allelic to Darier's disease. AB - Acrokeratosis verruciformis of Hopf is a localized disorder of keratinization affecting the distal extremities. Onset is early in life and the disease is inherited in an autosomal dominant fashion. Although histology of acrokeratosis verruciformis lesions shows no evidence of dyskeratosis, a possible relationship with Darier's disease has long been postulated on the basis of clinical similarity. ATP2A2 encoding the sarco(endo)plasmic reticulum Ca2+ ATPase2 pump has been identified as the defective gene in Darier's disease. In this report, we studied a family affected with acrokeratosis verruciformis in six generations and identified a heterozygous P602L mutation in ATP2A2. This mutation predicts a nonconservative amino acid substitution in the ATP binding domain of the molecule. The mutation segregates with the disease phenotype in the family and was not found in 50 controls. Moreover, functional analysis of the P602L mutant showed that it has lost its ability to transport Ca2+. This result demonstrates loss of function of the sarco(endo)plasmic reticulum Ca2+ ATPase2 mutant in acrokeratosis verruciformis, thus providing evidence that acrokeratosis verruciformis and Darier's disease are allelic disorders. PMID- 12542528 TI - Coupling of contact sensitizers to thiol groups is a key event for the activation of monocytes and monocyte-derived dendritic cells. AB - Strong contact sensitizers are able to induce distinct signal transduction mechanisms in antigen-presenting cells by coupling to cell proteins. The predominant target structures of haptens are thought to be thiol and amino groups in cysteine and lysine residues. We studied whether coupling of small reactive chemicals to thiol or amino groups might be responsible for the activation of monocytes and mature monocyte-derived dendritic cells. Human peripheral blood mononuclear cells were stimulated in vitro with subtoxic concentrations of the strong haptens 5-chloro-2-methylisothiazolinone plus 2-methylisothiazolinone and 2, 4, 6-trinitrochlorobenzene, the thiol-reactive reagents N-hydroxymaleimide and N-ethylmaleimide, as well as the amino-reactive compounds sulfosuccinimidyl acetate and 2-iminothiolane. Flow cytometric quantification of tyrosine phosphorylation in CD14+ monocytes showed that 5-chloro-2-methylisothiazolinone plus 2-methylisothiazolinone, 2, 4, 6-trinitrochlorobenzene, N-hydroxymaleimide, and N-ethylmaleimide but not sulfosuccinimidyl acetate and 2-iminothiolane strongly induced this process. Tyrosine phosphorylation induced by 5-chloro-2 methylisothiazolinone plus 2-methylisothiazolinone and 2, 4, 6 trinitrochlorobenzene was completely prevented in the presence of cysteine but not lysine, suggesting a competitive mechanism between cysteine and sulfhydryl groups of cell proteins. Using the mouse ear swelling test N-hydroxymaleimide could be classified as a significant contact allergen in comparison to 2, 4, 6 trinitrochlorobenzene, whereas no sensitizing potential became apparent for sulfosuccinimidyl acetate and 2-iminothiolane. Western blot analysis on monocytes and mature monocyte-derived dendritic cells confirmed the flow cytometric data for tyrosine phosphorylation and demonstrated a selective capacity of haptens and thiol-reactive compounds to activate ERK1/2 mitogen-activated protein kinase. Our data show that strong affinity of a small reactive chemical toward thiol groups is important for the activation of monocytes and monocyte-derived dendritic cells and can support the process of sensitization. PMID- 12542529 TI - Retinoic acid inhibits in vitro development of mast cells but has no marked effect on mature human skin tryptase- and chymase-positive mast cells. AB - Stem cell factor plays a key role in the development of human mast cells via interaction with Kit receptor. We and other groups have previously shown that a number of cytokines can regulate the stem-cell-factor-dependent development of mast cells in vitro. In this study we investigated the effect of retinoic acid on human mast cells in vitro and in vivo. Retinoids are known to have strong modulatory effects on hematopoietic differentiation. We found that all-trans retinoic acid, at concentrations as low as 1 nM, inhibits the stem-cell-factor dependent differentiation of mast cells in vitro. This effect of retinoic acid was found to be on progenitor cells, whereas more mature mast cells were less affected. The use of specific agonists binding either to the RAR or the RXR nuclear receptors indicated involvement of both the RAR/RXR and RXR/RXR pathways in inhibiting mast cell differentiation. In contrast to the effects on mast cell progenitors, retinoic acid had no effect on the number of mature mast cells in skin organ cultures. Furthermore, topical treatment of normal skin with a retinoic-acid-containing cream caused an increase in the number of tryptase positive mast cells, whereas the numbers of the major cutaneous mast cell type, tryptase- and chymase-positive mast cells, remained unaffected. Our results suggest that retinoic acid suppresses commitment of progenitor cells into the mast cell lineage and/or acts on early mast cell progenitors, whereas mature cutaneous mast cells are less susceptible to retinoic acid. PMID- 12542530 TI - Liver X receptor activators display anti-inflammatory activity in irritant and allergic contact dermatitis models: liver-X-receptor-specific inhibition of inflammation and primary cytokine production. AB - Activators of liver X receptors (LXR) stimulate epidermal differentiation and development, but inhibit keratinocyte proliferation. In this study, the anti inflammatory effects of two oxysterols, 22(R)-hydroxy-cholesterol (22ROH) and 25 hydroxycholesterol (25OH), and a nonsterol activator of LXR, GW3965, were examined utilizing models of irritant and allergic contact dermatitis. Irritant dermatitis was induced by applying phorbol 12-myristate-13-acetate (TPA) to the surface of the ears of CD1 mice, followed by treatment with 22ROH, 25OH, GW3965, or vehicle alone. Whereas TPA treatment alone induced an approximately 2-fold increase in ear weight and thickness, 22ROH, 25OH, or GW3965 markedly suppressed the increase (greater than 50% decrease), and to an extent comparable to that observed with 0.05% clobetasol treatment. Histology also revealed a marked decrease in TPA-induced cutaneous inflammation in oxysterol-treated animals. As topical treatment with cholesterol did not reduce the TPA-induced inflammation, and the nonsterol LXR activator (GW3965) inhibited inflammation, the anti inflammatory effects of oxysterols cannot be ascribed to a nonspecific sterol effect. In addition, 22ROH did not reduce inflammation in LXRbeta-/- or LXRalphabeta-/- animals, indicating that LXRbeta is required for this anti inflammatory effect. 22ROH also caused a partial reduction in ear thickness in LXRalpha-/- animals, however (approximately 50% of that observed in wild-type mice), suggesting that this receptor also mediates the anti-inflammatory effects of oxysterols. Both ear thickness and weight increased (approximately 1.5-fold) in the oxazolone-induced allergic dermatitis model, and 22ROH and GW3965 reduced inflammation by approximately 50% and approximately 30%, respectively. Finally, immunohistochemistry demonstrated an inhibition in the production of the pro inflammatory cytokines interleukin-1alpha and tumor necrosis factor alpha in the oxysterol-treated sites from both TPA- and oxazolone-treated animals. These studies demonstrate that activators of LXR display potent anti-inflammatory activity in both irritant and allergic contact models of dermatitis, requiring the participation of both LXRalpha and LXRbeta. LXR activators could provide a new class of therapeutic agents for the treatment of cutaneous inflammatory disorders. PMID- 12542531 TI - Macrophage colony-stimulating factor in cooperation with transforming growth factor-beta1 induces the differentiation of CD34+ hematopoietic progenitor cells into Langerhans cells under serum-free conditions without granulocyte-macrophage colony-stimulating factor. AB - Macrophage colony-stimulating factor has not been considered as a factor responsible for dendritic cell or Langerhans cell development from hematopoietic progenitor cells. In this study, we examined whether macrophage colony stimulating factor could be used instead of granulocyte-macrophage colony stimulating factor for the in vitro development of Langerhans cells from hematopoietic progenitor cells. We replaced granulocyte-macrophage colony stimulating factor with macrophage colony-stimulating factor from a serum-free culture containing granulocyte-macrophage colony-stimulating factor, stem cell factor, Flt3 ligand, tumor necrosis factor-alpha, and transforming growth factor beta1. This serum-free culture medium containing macrophage colony-stimulating factor, but not granulocyte-macrophage colony-stimulating factor (macrophage colony-stimulating factor culture), could induce CD1a+ Birbeck granule+ Langerin+ E-cadherin+ factor-like XIIIa Langerhans cells. As a control, the culture of hematopoietic progenitor cells in this culture medium depleted of macrophage colony-stimulating factor or transforming growth factor-beta1 resulted in far fewer or null CD1a+ cells, respectively. Macrophage colony-stimulating factor increased the number of CD1a+ cells in a concentration-dependent fashion. These macrophage colony-stimulating factor-induced Langerhans cells were different from granulocyte-macrophage colony-stimulating factor-induced Langerhans cells in their decreased expression of CD11c and their immature phenotype. The decreased expression of CD11c, however, was recovered by culturing them with granulocyte macrophage colony-stimulating factor, while they acquired a mature phenotype qby granulocyte-macrophage colony-stimulating factor, tumor necrosis factor-alpha, interleukin-1alpha, or lipo-polysaccharide. Macrophage colony-stimulating factor induced Langerhans cells could stimulate allogeneic T cells. Interestingly, we could keep the growth and immature phenotypes of macrophage colony-stimulating factor-induced Langerhans cells for at least 28 d of culture. These studies demonstrated that macrophage colony-stimulating factor in cooperation with transforming growth factor-beta1 could induce Langerhans cell development from hematopoietic progenitor cells in vitro without granulocyte-macrophage colony stimulating factor, which suggests the possibility that macrophage colony stimulating factor plays a part in the Langerhans cell development in vivo. In addition, the culture using macrophage colony-stimulating factor presents a novel culture system to enable a large-scale and long-term culture of immature Langerhans cells. PMID- 12542532 TI - Visualization and characterization of migratory Langerhans cells in murine skin and lymph nodes by antibodies against Langerin/CD207. AB - Dendritic cells are professional antigen-presenting cells that initiate primary immunity. Migration from sites of antigen uptake to lymphoid organs is crucial for the generation of immune responses. We investigated the migratory pathways specifically of epidermal Langerhans cells by tracing them from the epidermis to the draining lymph nodes. This was possible with a new monoclonal antibody, directed against murine Langerin/CD207, a type II lectin specific for Langerhans cells. In situ, resident, and activated Langerhans cells express Langerin in the epidermis and on their way through dermal lymphatic vessels. Both emigrated and trypsinization-derived Langerhans cells expressed high levels of Langerin intracellularly but reduced it upon prolonged culture periods. Sizeable numbers of Langerin+ cells were found in skin draining lymph nodes but not in mesenteric nodes. Langerin+ cells localized to the T cells areas and rarely to B cell zones. Numbers of Langerin-expressing cells increased after application of a contact sensitizer. In the steady state, Langerhans cells in the skin-draining nodes expressed maturation markers, such as 2A1 and costimulatory molecules CD86 and CD40. These molecules, CD86 and CD40, were further upregulated upon inflammatory stimuli such as contact sensitization. Thus, the novel anti-Langerin monoclonal antibody permits the unequivocal visualization of migratory Langerhans cells in the lymph nodes for the first time and thereby allows to dissect the relative immunogenic or tolerogenic contributions of Langerhans cells and other types of dendritic cells. PMID- 12542533 TI - Hydration disrupts human stratum corneum ultrastructure. AB - Using transmission and cryo-scanning electron microscopy, we confirm that extended water exposure leads to extensive disruption of stratum corneum intercellular lipid lamellae. We define the in vivo swelling behavior of the stratum corneum: exposure to water for 4 or 24 h results in a 3- or 4-fold expansion of the stratum corneum thickness, respectively. Corneocytes swell uniformly with the exception of the outermost and inner two to four corneocyte layers, which swell less. We show that hydration induces large pools of water in the intercellular space, pools that can exceed the size of water-swollen corneocytes. By 4 h of water exposure there are numerous small and large intercellular pools of water ("cisternae") present throughout the stratum corneum, and at 24 h these cisternae substantially increase in size. Within cisternae the lipid structure is disrupted by lamellar delamination ("roll-up"). Cisternae appear to be disk-shaped structures that do not obviously communicate. Cisternae appear to contain considerable lipidic and other material and to contain a substantial fluid volume that can rival the volume of the dry stratum corneum. Similar results are obtained following urine exposure. With urine exposure, cisternae communicate with salts in the external solution. This study illustrates the disruptive effect of overhydration on the stratum corneum intercellular space, identifies large and numerous unanticipated intercellular cisternal structures, defines the magnitude of stratum corneum swelling, and identifies stratum corneum cell layers that swell less. The study suggests the stratum corneum is a more chaotic structure than previously envisioned, and provides a framework for better understanding desquamation, irritancy, and percutaneous transport. PMID- 12542534 TI - Active influx transport is mediated by members of the organic anion transporting polypeptide family in human epidermal keratinocytes. AB - Normal human epidermal keratinocytes have been shown to express a cell-type specific pattern of extrahepatic cytochrome P450 enzymes and efflux transport proteins showing that these cells metabolize and excrete a variety of xenobiotics. Recently transport proteins involved in the uptake of xenobiotics have been detected and here we analyzed the mRNA and protein expression profiles and functional activities of these proteins in human keratinocytes in comparison to primary liver cells. The transporters studied included the subtypes A, B, C, D, and E of the organic anion transporting polypeptide (OATP) family, which are responsible for the uptake of various anionic and neutral molecules and especially organic cations - including drugs. Constitutive expression of OATP-B, OATP-D, and OATP-E was shown for the first time in normal human epidermal keratinocytes on a molecular level using reverse transcription polymerase chain reaction and northern blot analysis, as well as in human skin tissue shown by tissue blot hybridization and immunohistochemistry. Expression of OATP-A and OATP C was not detected in any of the keratinocyte samples. In contrast, liver tissue showed a significant expression of OATP-A and OATP-B as well as OATP-C, a weak expression of OATP-D, and no expression of OATP-E. These data revealed that normal human epidermal keratinocytes express a specific profile of transporters involved in drug influx. Using a newly developed uptake-transport assay, uptake of known and well-characterized OATP substrates like estradiol-17beta-glucuronide and estrone sulfate was inhibited in normal human epidermal keratinocytes by specific inhibitors such as taurocholate, verifying the functional capacity of the expressed OATPs. Human dermal fibroblasts seem to have a lower influx transport activity for estradiol-17beta-glucuronide, which correlates with the immunohistologic data. Even though the substrate specificity of the OATP isoforms is only partially known until now, our findings support the concept that uptake of large organic cations like drugs in keratinocytes is an active transport process mediated by members of the OATP family. PMID- 12542536 TI - Transcriptional regulation of the elafin gene in human keratinocytes. AB - Elafin (also known as skin-derived anti-leukoproteinase/trappin-2) is an epithelial host-defense protein that is absent in normal skin but highly induced in keratinocytes of inflamed skin (e.g., psoriasis), in epidermal skin tumors, and after wounding. Previously, it was shown that in cultured keratinocytes, elafin expression is induced by serum or tumor necrosis factor-alpha, and that expression is suppressed by retinoids, dithranol, and p38 mitogen-activated protein kinase inhibitors. Here we have studied the regulation of elafin gene expression in epidermal keratinocytes at the molecular level. First we determined the transcription start site of the elafin gene and found that the elafin mRNA possesses an unusually short 5'-untranslated region. Using transient transfection of luciferase reporter constructs of the elafin promoter, we mapped a 440 bp region upstream of the translation start site that conferred high-level expression in keratinocytes, but not in A431 cells or cells of mesenchymal origin. We observed that the promoter constructs were not subjected to the same regulation as the endogenous elafin gene as these constructs were highly active independent of keratinocyte stimulation. When elafin promoter constructs were stably transfected in the HaCaT keratinocyte cell line, tumor necrosis factor alpha inducible expression of both the endogenous elafin gene and the transgene was observed, suggesting that regulation of the elafin gene is also dependent on chromatin structure. We found, however, that a stably transfected 4 kb elafin promoter fragment did not confer retinoid sensitivity indicating that additional sequences are required for proper regulation. This study reveals the complex regulation of a gene that can be used as a paradigm for the specific differentiation program of activated epidermal keratinocytes. PMID- 12542535 TI - Cutaneous metabolism of vitamin B-6. AB - Vitamin B-6 is important for skin development and maintenance. We examined vitamin B-6 metabolism in human and mouse skin collected at different phases of the hair cycle; in hamster melanomas; in normal and immortalized human keratinocytes (HaCaT) and several human melanoma cell lines. Pyridoxamine 5' phosphate content was higher in mouse and hamster than in human skin. Activity of both pyridoxamine 5'-phosphate oxidase and pyridoxal 5'-phosphate hydrolase was significantly increased in rapidly growing melanomas compared to either normal skin or slower growing skin tumors. Reducing the pyridoxine content of the culture medium significantly increased the activity of pyridoxal kinase and pyridoxamine 5'-phosphate oxidase. Pyridoxal 5'-phosphate hydrolase has been proposed as a regulatory enzyme for vitamin B-6, but we found B-6 vitamer content to be significantly correlated only with kinase and oxidase activity and not with pyridoxal 5'-phosphate hydrolase activity. Although pyridoxal 5'-phosphate hydrolase activity is usually attributed to tissue-nonspecific alkaline phosphatase, tissue-nonspecific alkaline phosphatase knockout mice showed preservation of normal histology of the skin and adnexal structures. Furthermore, expression of tissue-nonspecific alkaline phosphatase mRNA was not detected in either HaCaT cells or human skin, both of which exhibited significant pyridoxal 5'-phosphate hydrolase activity. This suggests that an enzyme different from the classical tissue-nonspecific alkaline phosphatase may perform cutaneous pyridoxal 5'-phosphate hydrolase activity. PMID- 12542537 TI - Keratinocyte responsive element 3: analysis of a keratinocyte-specific regulatory sequence in the 230-kDa bullous pemphigoid antigen gene promoter. AB - The 230-kDa bullous pemphigoid antigen gene is expressed primarily, if not exclusively, in basal keratinocytes of the epidermis. Keratinocyte responsive element 3, a cis-element at position -216 to -197 of the human 230-kDa bullous pemphigoid antigen gene promoter, confers tissue-specific expression to this gene (Tamai et al: J Biol Chem 270:7609-7614, 1995). In this study, we investigated the functional characteristics of keratinocyte responsive element 3 on the 230 kDa bullous pemphigoid antigen gene core promoter by transient transfections of cultured normal human keratinocytes and normal human fibroblasts, as well as of lung carcinoma (A549), osteosarcoma (OST), and gastric adenocarcinoma (GT3TKB) cell lines. A 230-kDa bullous pemphigoid antigen gene core promoter/luciferase reporter gene plasmid construct, pBPL, was modified to develop a series of constructs (pKBPL-p4KBPL), which have insertions of one, two, three, or four tandem repeats of keratinocyte responsive element 3, and these plasmids were used in transient transfections of the cultured cells. The promoter activities of pKBPL-p4KBPL constructs, relative to pBPL, in normal human keratinocytes were 7.6 , 15.5-, 4.6-, and 2.7-fold higher, respectively, whereas no upregulatory effect by keratinocyte responsive element 3 insertion was observed in other cell lines tested. prKBPL, a plasmid constructed with keratinocyte responsive element 3 in reverse orientation, showed essentially no activity in normal human keratinocytes. Insertion of a random 20 bp sequence between keratinocyte responsive element 3 and the 230-kDa bullous pemphigoid antigen gene core promoter resulted in about 40% reduction of luciferase activity in normal human keratinocytes. These data suggest that keratinocyte responsive element 3 functions as a position-, copy number-, and orientation-dependent cis-element contributing to tissue-specific regulation of the 230-kDa bullous pemphigoid antigen gene. PMID- 12542538 TI - Normal human Merkel cells are present in epidermal cell populations isolated and cultured from glabrous and hairy skin sites. AB - The Merkel cell is a highly specialized cell that primarily acts as a slowly adapting mechanoreceptor. Merkel cells are scarce in normal skin but can be identified by the expression of distinct keratin filaments. Merkel cells constitute a very unique population and many questions still remain as to their origin, number, proliferative capacity, and functions in cutaneous biology. The dissociation of epidermal cells from skin is a widely used technique to extract and culture keratinocytes. We took advantage of a two-step extraction method to quantify keratin-20-expressing Merkel cells among total cutaneous cells obtained from either hairy or glabrous skin biopsies. Flow cytometry analysis revealed that keratin-20-labeled Merkel cells represent between 3.6% and 5.7% of freshly dissociated basal epidermal cells. No significant differences were seen between samples derived from glabrous palmar and hairy anatomic sites, from children and adult, respectively. We also report on the presence of Merkel cells in primary and first subcultures of epidermal cells indicating their capacity to remain viable after extraction from skin of various anatomic sites. To our knowledge, this is the first demonstration of nontumorigenic human Merkel cells in culture in vitro. The persistence of a small number of Merkel cells in culture suggests that, with the development of appropriate culture conditions, these cells could be amplified and further studied to unravel long-standing questions relative to their paracrine function or epithelial origin. PMID- 12542539 TI - Induction of metallothionein in human skin by routine exposure to sunlight: evidence for a systemic response and enhanced induction at certain body sites. AB - Expression of metallothionein, an antioxidant induced by a variety of stimuli including ultraviolet light, was quantitated by immunohistochemistry in the skin of males aged over 50 who had known short- and long-term exposures to sunlight. Skin punch biopsies were taken from two sites in each subject: the hand in all subjects and a range of other sites matched to patients with a previously excised primary melanoma. Metallothionein expression (strongest in the basal layers of the epidermis and primarily nuclear) was associated with both short- and long term exposure to sunlight. A plateau of staining intensity was reached after 3 h sun exposure, within the previous 3 d before biopsy. Expression was also elevated in the nonexposed skin sites of subjects who had recent sun exposure, indicating a systemic response to exposure of remote sites. Using the skin of the hand to normalize responses to chronic exposure between individuals, the systemically modulated response to sunlight was significantly greater on the unexposed back than on other sites. The possibility of ultraviolet-induced cytokines selectively modifying the response of skin on a site-specific basis was investigated. The circulating leukocytes, but not lymphocytes, of two individuals exposed to 1 minimal erythema dose whole-body solar-simulated ultraviolet showed increased interleukin-6 mRNA 4 h after exposure. Interleukin-6 was not directly induced in these cell populations 4 h after ultraviolet A or ultraviolet B irradiation ex vivo. Leukocytes may therefore contribute to and amplify the systemic effects of ultraviolet-induced interleukin-6 and metallothionein expression. PMID- 12542540 TI - Influence of 5-aminolevulinic acid and red light on collagen metabolism of human dermal fibroblasts. AB - Patients with localized scleroderma receiving topical photodynamic therapy with 5 aminolevulinic acid show a reduction in skin tightness, suggesting that this therapy reduces skin sclerosis. To investigate potential mechanisms, the effects of 5-aminolevulinic acid and light on collagen metabolism were studied in vitro. Normal and scleroderma fibroblasts were treated with sublethal doses of 5 aminolevulinic acid and red light and transferred to three-dimensional collagen lattices. Cell supernatants were taken 6-72 h after photodynamic therapy to determine protein levels of the matrix metalloproteinases 1, 2, and 3, and of their inhibitors, tissue inhibitor of metalloproteinase 1 and 2 by enzyme-linked immunosorbent assay. Cellular mRNA expression of these proteins and of collagen type I and III was measured by quantitative real-time polymerase chain reaction. A significant, time-dependent induction of matrix metalloproteinase 1 (up to 2.4 fold after 48 h) and matrix metalloproteinase 3 (up to 4.3-fold after 48 h) protein levels was seen after 5-aminolevulinic acid-photodynamic therapy. Irradiation with ultraviolet A light, used as a positive control, showed a similar induction of matrix metalloproteinase 1 (2.3-fold after 48 h). The mRNA levels of matrix metalloproteinase 1 and matrix metalloproteinase 3 were significantly increased 12 h after irradiation, whereas collagen type I mRNA was strongly decreased already 6 h following irradiation. Collagen type III, tissue inhibitor of metalloproteinase 1, and matrix metalloproteinase 2 did not change after photodynamic therapy. Addition of nontoxic concentrations of sodium azide, a singlet-oxygen quencher, significantly inhibited induction of matrix metalloproteinase 1 by 5-aminolevulinic acid and light. These data show that 5 aminolevulinic acid and light induce matrix metalloproteinase 1 and matrix metalloproteinase 3 expression in normal and scleroderma fibroblasts in a singlet oxygen-dependent way while reducing collagen type I mRNA expression. Induction of collagen-degrading enzymes together with reduction of collagen production might be responsible for the anti-sclerotic effects of 5-aminolevulinic acid photodynamic therapy observed in vivo. PMID- 12542541 TI - Endothelium-dependent regulation of cutaneous microcirculation in patients with systemic scleroderma. AB - The microinjection method used for the first time in this study makes it possible to measure in vivo the endothelium-dependent vasomotion in the nailfold capillaries of systemic scleroderma patients. Aceylcholine (blood concentration of 10(-5) M) and sodium nitroprusside (blood concentration of 1.6 microg per kg per min) were used as test substances to examine capillary vasomotion. Defined quantities of medication in the range of microliters were administered as a bolus using the Panomat V-3 medicine pump. The boluses were injected into the capillaries at the capillary loop in 10 scleroderma patients and 10 randomized healthy volunteers. Blood flow velocity and the diameter of the skin capillaries at the venous loop were measured and tested for correlation with the effects of injections of test substances. The intracapillary administration of sodium nitroprusside brought about a significant decrease in capillary blood flow velocity and a significant increase in vascular diameter in comparison with baseline values, both in systemic scleroderma patients (p<0.005 and p<0.0001) and in the healthy volunteers (p<0.001 and p<0.0001). The intracapillary administration of acetylcholine led to a significant decrease in blood flow velocity (p<0.001) and increase in vascular diameter (p<0.0001) only in the healthy volunteers. In systemic sclerosis patients capillary diameter and blood flow velocity were unaffected. In two systemic scleroderma patients this procedure was repeated in identical fashion at the end of a 1 wk infusion therapy cycle with prostacyclin. The infusion therapy led to a normalization of endothelium-dependent vasomotion in the two scleroderma patients. The microinjection technique used in this study for the first time made it possible to establish in vivo that endothelium-dependent vasomotion is disturbed in systemic scleroderma patients. PMID- 12542542 TI - Neutralization of hepatocyte growth factor leads to retarded cutaneous wound healing associated with decreased neovascularization and granulation tissue formation. AB - To elucidate biologic functions of hepatocyte growth factor and the c-Met receptor in cutaneous wound healing, we analyzed expression and localization of hepatocyte growth factor and c-Met receptor and used a strategy to neutralize endogenous hepatocyte growth factor in a cutaneous wound healing model in mice. Following excision of full-thickness skin on the dorsum of mice, expression of both hepatocyte growth factor and the c-Met receptor increased transiently in cutaneous tissues. Expressions of hepatocyte growth factor increased as early as 2 d postwounding and reached a peak on day 2, whereas the c-Met receptor expression reached a peak 2-4 d postwounding. Immunolocalization of the c-Met receptor indicated that c-Met receptor expression was upregulated in keratinocytes, vascular endothelial cells, and myofibroblasts in granulation tissue, hence these are potential target cells of hepatocyte growth factor. When normal rabbit IgG or neutralizing anti-hepatocyte growth factor IgG was locally and continuously delivered to subcutaneous lesions, the number of capillary vessels decreased with the neutralization of hepatocyte growth factor and there was an associated decreased expansion of granulation tissue. Likewise, retardation in re-epithelialization and the rate of wound closure occurred with neutralization of endogenous hepatocyte growth factor on days 4 and 7 postwounding. Therefore, hepatocyte growth factor is definitely involved in enhancing cutaneous wound healing processes, including re-epithelialization, neovascularization, and granulation tissue formation. PMID- 12542544 TI - Clinical snippets. PMID- 12542545 TI - In this issue: genotype-phenotype correlations: lessons learned from 'milder' genodermatoses. PMID- 12542546 TI - An orphan meets family members in skin. PMID- 12542547 TI - Assessing the safety of immunologic modifiers for the treatment of chronic disease: the psoriasis paradigm. PMID- 12542548 TI - Paracetamol hepatotoxicity at therapeutic doses. PMID- 12542549 TI - The risk of thrombolysis in association with cardiopulmonary resuscitation: no reason to withhold this causal and effective therapy. PMID- 12542550 TI - Helicobacter pylori: resurrection of the cancer link. AB - Helicobacter pylori is one of the most common pathogenic bacterial infections, colonizing an estimated half of all humans. In a subset of individuals, the infection leads to serious gastroduodenal disease such as peptic ulcers and gastric adenocarcinoma. The factors contributing to skewing this, in most cases benign, relationship into disease development are largely unknown. However, factors emanating from the bacterium, host and the environment have been shown to affect the risk for disease, although no factor can be singled out to be most important. The known factors are associated with affecting the risk of disease, and are not absolute. Virulence of H. pylori is affected by the existence and regulation of certain genes present in the bacterial population in a stomach. The effects of H. pylori on gastric cancer development have been challenged and the risk associated with infection with virulent (i.e. Cag PAI positive) H. pylori has likely been underestimated. PMID- 12542551 TI - The molecular genetics of cardiovascular disease: clinical implications. AB - Coronary heart disease (CHD) is one of the main leading causes of death worldwide. CHD is a complex condition resulting from numerous gene-gene and gene environment interactions. A large number of candidate genes exist. We review the evidence for the role of apolipoprotein E, lipoprotein lipase and interleukin-6 in CHD and their interaction with smoking (an environmental risk). The main objective of genetic research into CHD is to provide a complete risk assessment, complementing the well-studied traditional clinical and biochemical risk factors. Unravelling the role that functional gene polymorphisms play in determining risk, and in determining the levels of intermediate phenotypes is crucial to our understanding of the key metabolic pathways and physiology not only in the diseased, but also in the disease-free state. The identification of new molecules (implicated in the relevant metabolic pathways) may subsequently lead to potential targets for therapeutic intervention. Improving our molecular understanding may also lead to the development of improved diagnostics. Sequencing of the entire genome is now a reality. Advances in technology, such as the development of "gene chips" may allow us to study 100 000 genes at a time on a single chip. Such advances will amplify the power available to us in the realms of research into genetic physiology and interaction with environmental stimuli. The insights that these novel approaches may yield are eagerly awaited. PMID- 12542552 TI - Major bleeding complications after cardiopulmonary resuscitation: impact of thrombolytic treatment. AB - OBJECTIVE: The risk of bleeding complications caused by thrombolysis in patients with cardiac arrest and prolonged cardiopulmonary resuscitation is unclear. We evaluate the complication rate of systemic thrombolysis in patients with out-of hospital cardiac arrest caused by acute myocardial infarction, especially in relation to duration of cardiopulmonary resuscitation. DESIGN: The study was designed as retrospective cohort study, the risk factor being systemic thrombolysis and the end-point major haemorrhage, defined as life-threatening and/or need for transfusion. Over 10.5 years, emergency cardiac care data, therapy, major haemorrhage and outcome of 265 patients with acute myocardial infarction admitted to an emergency department after successful cardiopulmonary resuscitation were registered. RESULTS: We observed major haemorrhage in 13 of 132 patients who received thrombolysis (10%, 95% confidence interval 5-15%), five of these survived to discharge, none died because of this complication. Major haemorrhage occurred in seven of 133 patients in whom no thrombolytic treatment had been given (5%, 95% confidence interval 1-9%), two of these survived to discharge. Taking into account baseline imbalances between the groups, the risk of bleeding was slightly increased if thrombolytics were used (odds ratio 2.5, 95% confidence interval 0.9-7.4) but this was not significant (P = 0.09). There was no clear association between duration of resuscitation and bleeding complications (z for trend = 1.52, P = 0.12). Survival was not significantly better in patients receiving thrombolysis (odds ratio 1.6, 0.9-3.0, P = 0.12). CONCLUSIONS: Bleeding complications after cardiopulmonary resuscitation are frequent, particularly in patients with thrombolytic treatment, but do not appear to be related to the duration of resuscitation. In the light of possible benefits on outcome, thrombolytic treatment should not be withheld in carefully selected patients. PMID- 12542553 TI - Hyperinsulinaemia as long-term predictor of death and ischaemic heart disease in nondiabetic men: The Malmo Preventive Project. AB - OBJECTIVES: Prospective studies have indicated that hyperinsulinaemia/insulin resistance is a risk factor for ischaemic heart disease (IHD), the risk decreasing with time of follow-up. Few studies have so far investigated the role of hyperinsulinaemia in the prediction of long-term total mortality. SETTING: Section of Preventive Medicine, Department of Medicine, University Hospital, Malmo, Sweden. SUBJECTS: A total of 6074 nondiabetic, middle-aged, healthy Swedish males. SCREENING EXAMINATION: We determined IHD risk factors including blood glucose and plasma insulin before and 2 h after an oral glucose tolerance test (OGTT). Total follow-up time was 19 years. Hyperinsulinaemia was defined as values above the 10th decentile of fasting or 2 h insulin concentration. MAIN OUTCOME MEASURES: Total mortality and cardiac event (CE) rate for IHD. RESULTS: Unadjusted relative risks (RRs) for both death and CE were J-shaped with the highest relative risk (RR: 1.4-1.6) in the hyperinsulinaemic group compared with all other men. The RRs for death and CE were significant for fasting insulin but became nonsignificant after adjustment for other risk factors and also with a longer follow-up. The risk of death in hyperinsulinaemic men, defined on the basis of 2-h insulin level, increased with time of follow-up and was still significantly increased after 19 years [RR: 1.32 (95% CI: 1.05-1.65], even after adjustment for other risk factors. CONCLUSIONS: Fasting hyperinsulinaemia was a predictor of total mortality and IHD in nondiabetic men, although not more significantly after adjustment for other risk factors and with lengthening of follow-up time. The 2-h postglucose hyperinsulinaemia appeared to be a stronger and independent predictor of mortality over long-term follow-up. These findings support the view that insulin resistance with associated cluster of risk factors predicts increased long-term risk of mortality and IHD. PMID- 12542554 TI - Vitamin B-12 treatment has limited effect on health-related quality of life among individuals with elevated plasma methylmalonic acid: a randomized placebo controlled study. AB - OBJECTIVE: To examine the hypothesis that treatment with vitamin B-12 improves health-related quality of life (HRQOL) in individuals with biochemical signs of vitamin B-12 deficiency. DESIGN: A randomized placebo-controlled study. SETTING: Municipality of Aarhus, Denmark. SUBJECTS: Nonhospitalized individuals (n = 140) with a modest increase in plasma methylmalonic acid (0.40-2.00 micromol L-1) not previously treated with vitamin B-12. INTERVENTION: The participants were randomized to vitamin B-12 injection treatment or placebo weekly for 4 weeks and re-examined 3 months later. The investigator and the participants were blinded to the intervention. MAIN OUTCOME MEASURE: Change in HRQOL assessed by the SF-36 questionnaire from baseline to follow-up examination 3 months later. RESULTS: The participants reported a significantly worser HRQOL than the age- and sex-matched Danish general population (P < 0.001). However, no change was observed after treatment with vitamin B-12 for seven of eight health dimensions. A significant improvement was found only in general health when compared with the placebo group (P = 0.03). CONCLUSIONS: Vitamin B-12 treatment influenced only one of eight dimensions of HRQOL amongst participants with biochemical signs of vitamin B-12 deficiency. We therefore question the benefit of vitamin B-12 treatment amongst elderly with a modestly increased plasma methylmalonic acid as the only sign of vitamin B-12 deficiency. PMID- 12542555 TI - Elevated cardiac troponin T in predialysis patients is associated with inflammation and predicts mortality. AB - OBJECTIVES: Cardiac troponin T (cTnT) is a highly sensitive and specific marker of myocardial damage. It has been shown that elevated serum concentrations of cTnT in haemodialysis (HD) patients are associated with poor prognostic outcome. The aim of the present study was to investigate the predictive value of cTnT in samples from predialysis patients and to investigate associations between cTnT and inflammatory markers, such as C-reactive protein (CRP) and interleukin-6 (IL 6). DESIGN: Cohort, follow-up study. SETTING: Huddinge University Hospital, Sweden. SUBJECTS: A total of 115 (62% males, 28% diabetic patients) end-stage renal disease (ESRD) patients (52 +/- 1 years), of which 29% had cardiovascular disease (CVD), were studied shortly before the onset of dialysis therapy. Sixty four patients started peritoneal dialysis (PD) as renal replacement therapy, whilst 49 started HD during the follow-up. MAIN OUTCOME MEASURES: The cTnT was analysed with the third generation TnT assay on Elecsys 2010. The prognostic value was calculated for cTnT, IL-6, age, CVD, malnutrition, diabetes mellitus (DM) and gender. Survival analyses were made with Kaplan-Meier and Cox regression analyses, with all-cause mortality as the clinical end point (mean follow-up period 2.7 +/- 0.1 years). RESULTS: Significant correlations were found between cTnT and CKMB (rho = 0.52, P < 0.0001), IL-6 (rho = 0.23, P < 0.05), CRP (rho = 0.30, P < 0.05), and serum albumin (rho = -0.31, P < 0.001), respectively. Diabetic patients had higher median serum cTnT level (0.09 microg L-1; range <0.01-0.51 vs. 0.04 microg L-1; range <0.01-0.67 microg L-1; P < 0.005) compared with nondiabetic patients. Likewise, patients with CVD had a significantly higher median level (0.08 microg L-1; range <0.01-0.67 microg L-1 vs. 0.04 microg L-1; range <0.01-0.61 microg L-1; P < 0.01) of cTnT compared with patients without CVD. Patients with cTnT > or =0.10 microg L-1 had a higher cumulative mortality rate than patients with cTnT < 0.10 microg L-1 (chi2 = 7.04; P < 0.01). Whilst age, CVD, malnutrition, DM, IL-6, cTnT and male gender were associated with poor outcome in the univariate analysis, only DM (P < 0.05) and cTnT (P < 0.05) were independently associated with mortality in a multivariate analysis. CONCLUSIONS: The present study demonstrates that serum concentrations of cTnT > or =0.10 microg L-1 is a significant predictor of mortality in patients starting dialysis. Moreover, the positive correlations between cTnT and IL-6, and CRP, respectively, suggest an association between inflammation and cTnT levels. Finally, the results of the present study suggest that cTnT is an independent predictor of mortality in ESRD patients starting dialysis. PMID- 12542556 TI - Prevalence and significance of cardiovascular risk factors in a large cohort of patients with familial hypercholesterolaemia. AB - OBJECTIVE: Patients with familial hypercholesterolaemia (FH) vary widely in terms of onset of cardiovascular disease (CVD). DESIGN: The association between cardiovascular risk factors and prevalent CVD was examined in a cross-sectional study in order to elucidate their contribution to atherogenesis. SETTING AND SUBJECTS: Patients were recruited from 37 Dutch Lipid Clinics. The diagnosis of FH was based on a uniform diagnostic protocol, confirmed by DNA analysis in 62% of the cases. All patients were investigated free from any lipid-lowering drug for at least 6 weeks. MAIN OUTCOME MEASURES: Differences in lipids, lipoproteins and other risk factors for CVD were analysed in FH patients with and without CVD. RESULTS: A total of 526 patients were assessed and more than 37% had a history of CVD with a mean age of onset of 46.8 years. Mean LDL cholesterol (LDL-C) levels were severely elevated (8.38 +/- 2.13 mmol L-1). In univariate analysis, age, presence of hypertension or diabetes, body mass index, triglycerides (TG) and low HDL cholesterol (HDL-C) were all significantly associated with CVD. Also in multivariate analysis, all these risk factors, except TG and diabetes, were significantly linked to CVD. CONCLUSION: A high CVD risk in this large well documented characterized sample of FH patients is not only conferred by elevated LDL-C but also by low HDL-C. PMID- 12542557 TI - What cause of mortality can we predict by cholesterol screening in the Japanese general population? AB - OBJECTIVE: In a population with a markedly lower coronary mortality such as in Japan, the benefit of cholesterol screening may be different from Western populations. We attempted to assess the importance of cholesterol screening in Japan. DESIGN: A 13.2-year cohort study for cause-specific mortality. SETTING: Three hundred randomly selected districts throughout Japan in which the National Survey on Circulatory Disorders 1980 was performed. SUBJECTS: A total of 9216 community dwelling persons aged 30 years and over, with standardized serum cholesterol measurement and without a past history of cardiovascular disease. RESULTS: There were 1206 deaths, which included 462 deaths due to cardiovascular disease with 79 coronary heart diseases. Hypercholesterolemia (>6.21 mmol L-1) showed a significant positive relation to coronary mortality (relative risk; 2.93, 95% confidence interval; 1.52-5.63) but not to stroke. Although hypocholesterolemia (<4.14 mmol L-1) was significantly associated with an increased risk of liver cancer, noncardiovascular, noncancer disease and all cause mortality, these associations, except for liver cancer, disappeared after excluding deaths in the first 5 years of the follow-up. The multivariate adjusted attributable risk of hypercholesterolaemia for coronary disease was 0.98 per 1000 person-years, which was threefold higher than that of hypocholesterolemia for liver cancer: 0.32 per 1000 person-years. The attributable risk percentage of hypercholesterolaemia was 66% for coronary heart disease. CONCLUSION: Similar to Western populations, it is recommended to provide screening for hypercholesterolaemia in Japan, especially for males, although its attributable risk for coronary disease might be small. PMID- 12542558 TI - Acupuncture versus placebo versus sumatriptan for early treatment of migraine attacks: a randomized controlled trial. AB - OBJECTIVES: To investigate whether acupuncture is superior to placebo and equivalent to sumatriptan for the early treatment of an acute migraine attack. DESIGN: Randomized, partly double-blind (sumatriptan versus placebo) trial. SETTING: Two hospitals in Germany (one specialized in traditional Chinese medicine and one in the treatment of headache). SUBJECTS: A total of 179 migraineurs experiencing the first symptoms of a developing migraine attack. INTERVENTIONS: Traditional Chinese acupuncture, sumatriptan (6 mg subcutaneously) or placebo injection. MAIN OUTCOME MEASURE: Number of patients in whom a full migraine attack (defined as severe migraine headache) within 48 h was prevented. In patients who developed a migraine attack in spite of early treatment, acupuncture and sumatriptan were applied a second time, whilst patients initially randomized to placebo received sumatriptan. RESULTS: A full migraine attack was prevented in 21 of 60 (35%) patients receiving acupuncture, 21 of 58 (36%) patients receiving sumatriptan and 11 of 61 (18%) patients receiving placebo (relative risk of having a full attack 0.79 (95% CI, 0.64-0.99) for acupuncture versus placebo, and 0.78 (95% CI, 0.62-0.98) for sumatriptan versus placebo). Response to the second intervention in patients who developed a full attack was better with sumatriptan (17/31 patients who received sumatriptan twice and 37/46 patients who had had placebo first) than with acupuncture (4/31). The number of patients reporting side-effects was 14 in the acupuncture group, 23 in the sumatriptan group and 10 in the placebo group. CONCLUSIONS: In this trial acupuncture and sumatriptan were more effective than a placebo injection in the early treatment of an acute migraine attack. When an attack could not be prevented, sumatriptan was more effective than acupuncture at relieving headache. PMID- 12542559 TI - Validation of a glucose-insulin-potassium infusion algorithm in hospitalized diabetic patients. AB - OBJECTIVES: Validation of a novel glucose-insulin-potassium (GIK) infusion algorithm to optimize metabolic control in hospitalized diabetic patients. SUBJECTS: We randomized 33 diabetic patients admitted to Sahlgrenska University Hospital with acute internal medicine diseases to either GIK infusion or multiple injection therapy (MIT). The GIK infusion rate and the MIT were controlled according to special algorithms. The treatment efficacy was evaluated through comparisons of capillary blood glucose eight times on day 2 of the study. RESULTS: The GIK infusion led to significantly lower mean blood glucose when compared with MIT [10.1 (9.0-11.2) vs. 12.3 (9.3-14.4) mmol L(-1), median and interquartile range, P < 0.01]. Four episodes of hypoglycaemia without loss of consciousness were recorded in the GIK group whereas no hypoglycaemic event occurred in the MIT group. A mean of 1 (1-3) episodes of blood glucose levels above 12.0 mmol L(-1) were recorded in the GIK group compared with 3.5 (1.5-5.0) in the MIT group, P < 0.01. CONCLUSIONS: The algorithm used for the GIK infusion gave an acceptable level of metabolic control and this insulin infusion protocol is safe enough to be used by the nursing staff on a general internal medicine ward. PMID- 12542560 TI - Association between vitamin D receptor gene polymorphisms and tubular citrate handling in calcium nephrolithiasis. AB - OBJECTIVES: Hypocitraturia is a risk factor for calcium nephrolithiasis. 1,25(OH)2D3 influences renal citrate handling and enhances citraturia. The aim of this study was to evaluate the relationship between vitamin D receptor (VDR) allelic variant and urinary citrate excretion in recurrent stone formers (SF) patients. DESIGN: Case-control study. SUBJECTS: A total of 220 recurrent calcium oxalate SF patients and 114 healthy control (C) subjects were enrolled for this study. Subjects with urinary tract infections, hyperparathyroidism, cystinuria >70 micromol/24 h, gouty diathesis, renal tubular acidosis, renal failure, chronic diarrhoeal states, intake of thiazide diuretics, angiotensin-converting enzyme (ACE)-inhibitors, glucocorticoids or oestrogens were excluded. A standard constant diet was given for 7 days. The 24-h urinary citrate excretion and the active tubular reabsorption of filtered citrate (Rcit) were evaluated. Hypocitraturia was defined as a urinary citrate excretion lower than 1.7 mmol day 1. Stone formers patients and C were genotyped for BsmI and TaqI VDR alleles. Contingency table chi-square tests were used to compare genotype frequencies in hypocitraturic SF patients, normocitraturic SF and C. RESULTS: The prevalence of hypocitraturia in SF patients was 32.7% (72 of 200). Hypocitraturia in these patients resulted from excessive Rcit of a normal load of citrate. We found a different distribution (P < 0.05) of BsmI and TaqI VDR genotypes in hypocitraturic SF patients compared with normocitraturic SF and C. In particular, the prevalence of bb and TT VDR genotypes in hypocitraturic SF was significantly higher than in normocitraturic SF and C. CONCLUSIONS: These results point to a genetic association between BsmI and TaqI VDR polymorphisms and idiopathic hypocitraturia in calcium-oxalate recurrent SF patients. PMID- 12542561 TI - No acute impact of haemodialysis treatment on free radical scavenging enzyme gene expression in white blood cells. AB - OBJECTIVE: Oxidative stress has been implicated in the side-effects caused by haemodialysis (HD) treatment. DESIGN: In the present study we have investigated whether gene expression of the enzymatic defence system provided by cellular glutathione peroxidase (GPx-1), phospholipid glutathione peroxidase (GPx-4), glutathione reductase (GSSG-R), glutathione synthethase (GSH-S), Cu/Zn-superoxide dismutase (SOD-1) and catalase (CAT) is affected by HD. The GPx-1, GPx-4, GSSG-R, GSH-S, SOD-1 and CAT mRNA were determined in white blood cells by quantitative reverse transcriptase-polymerase chain reaction with the LightCycler instrument and transcription elongation factor-2 as reference gene at the start (SD) and immediately after (ED) dialysis treatment (n = 36). In a subgroup (n = 10), messenger RNA (mRNA) expression was determined hourly during a 5 h HD. RESULTS: The expression of GPx-1, GPx-4, GSSG-R, GSH-S, SOD-1 and CAT mRNA was not affected by a single HD treatment. All mRNAs were significantly (P < 0.05) increased in HD patients [median (16. percentiles (perc.); 84. perc.)]: GPx-1: 2.18 (0.89; 3.23); GPx-4: 0.41 (0.26; 0.74); GSSG-R: 0.04 (0.02; 0.10); GSH-S: 0.04 (0.02; 0.08); SOD-1: 0.32 (0.20; 0.62); CAT: 0.12 (0.06; 0.18) when compared with healthy blood donors (GPx-1: 0.91 (0.60; 1.44); GPx-4: 0.27 (0.16; 0.43); GSSG-R: 0.02 (0.01; 0.02); GSH-S: 0.02 (0.02; 0.04); SOD-1: 0.15 (0.10; 0.18); CAT: 0.07 (0.04; 0.16). CONCLUSIONS: These results show that the HD procedure does not acutely affect the antioxidant defence system on the gene level but suggest that the chronic stress caused by uraemia and/or HD may cause gene induction of the enzymatic defence system. PMID- 12542562 TI - Phospholipid and cholesteryl ester transfer are increased in lipoprotein lipase deficiency. AB - OBJECTIVES: Phospholipid transfer protein (PLTP) and cholesteryl ester transfer protein (CETP) are key enzymes in lipoprotein metabolism by mediating the transfer and exchange of phospholipids (PL) and neutral lipids between lipoproteins. Lipoprotein lipase (LPL) deficiency is associated with low HDL cholesterol (HDL-C) levels in both, the homozygous and heterozygous state. In the present study we set out to investigate the role of lipid transfer proteins, which are known to strongly determine HDL-C levels, in LPL deficiency. DESIGN/SUBJECTS: Phospholipid acceptor and donor properties of lipoproteins, PLTP activity, CETP mass, activity and cholesteryl ester (CE) transfer were determined in two homozygous and six heterozygous LPL-deficient subjects and in 10 healthy, normolipidaemic controls, respectively. RESULTS: The HDL isolated from LPL deficient subjects showed strongly increased PL-acceptance when compared with controls (homozygotes versus heterozygotes versus control: 26.46 +/- 15.26 vs. 3.41 +/- 1.61 vs. 1.89 +/- 0.33 micromol mL-1 h-1/micromol mL-1 PL; all P < 0.05). Phospholipid transfer from apolipoprotein B containing lipoproteins was increased in heterozygotes when compared with controls (46.66 +/- 23.3 vs. 28.91 +/- 18.05 micromol mL-1 h-1/micromol mL-1 PL, P = 0.05). PLTP activity, however, was similar in LPL-deficient subjects and controls. CETP mass was highest in homozygotes, whilst enzyme activity was similar in LPL-deficient subjects and controls. CE transfer was highest in homozygotes (72.5 +/- 8.8%) and lowest in controls (28.7 +/- 5.2%, P < 0.01). CONCLUSIONS: In conclusion, PL and CE transfer are increased in LPL deficiency and thus, partly explain low HDL-levels in LPL-deficient subjects. Enhanced transfer seems rather to be the result of altered lipoprotein composition and concentration than altered enzyme activity. Our findings on mechanisms leading to low HDL-C levels might show another aspect in atherogenesis in LPL deficiency. PMID- 12542563 TI - A targeted approach significantly increases the identification rate of patients with undiagnosed haemochromatosis. AB - OBJECTIVE: To determine the optimal means of identifying patients with undiagnosed haemochromatosis. DESIGN: Case-control study where cases are defined by the presence of specific clinical diagnoses or symptoms. SETTING: Primary care patients were recruited from three Oxfordshire practices and secondary care patients were recruited from those patients attending specialist clinics in Amiens University Hospital. SUBJECTS: A total of 569 patients recruited via hospital clinics and 60 primary care patients (recruited from 4022 consultations) presenting with the following haemochromatosis associated conditions, diabetes, arthralgia/chronic fatigue, osteoporosis or arthropathy were studied. The control group, a total of 991 healthy volunteers, were recruited through a Health Appraisal Centre. Patients and controls were included in the study if they or their family members had not previously been diagnosed with hereditary haemochromatosis. MAIN OUTCOME MEASURES: Serum ferritin concentration, transferrin saturation (Tsat) and presence of HFE mutations, C282Y and H63D. The check-up in controls consisted of a questionnaire, clinical examination, biochemical tests and screening for the presence of the C282Y and H63D mutations. RESULTS: Patient groups presenting with unstable diabetes or chronic fatigue and arthralgia together with a raised serum ferritin concentration showed an enrichment in the haemochromatosis-associated genotype HH/YY, odds ratio (OR) = 40.1, confidence interval (CI) = 8.0-202.1 and OR = 103, CI = 22.9-469.7, respectively. CONCLUSION: Patients presenting to hospital clinics with haemochromatosis associated conditions should be screened biochemically for iron overload. Only those with a serum ferritin >300 microg L-1 or Tsat >40% should subsequently go on to be genotyped for HFE mutations. The patients at greatest risk of having undiagnosed haemochromatosis are those presenting with unstable diabetes, or fatigue and/or arthralgia in the absence of any other explanation. PMID- 12542564 TI - Effects of glucocorticoids on leptin levels and eating behaviour in women. AB - OBJECTIVE: Long-term treatment with glucocorticoids induces weight gain and increased risk to develop obesity-related metabolic complications. The underlying mechanisms are not fully understood. Glucocorticoid therapy has previously been associated with increased levels of circulating leptin. In this study the eating behaviour was therefore studied in relation to leptin levels before and after short-term prednisolone treatment. DESIGN: Within-subject design. SUBJECTS: Twelve healthy postmenopausal women with a mean body mass index (BMI) of 28.9 kg m-2 (+/-0.8 SEM) volunteered after recruitment by an advertisement in the local paper. INTERVENTIONS: The subjects received 25 mg prednisolone daily for 7 days. MAIN OUTCOME MEASUREMENTS: Fasting serum samples were obtained before, during and after treatment for determination of leptin and insulin, glucose and fractionated lipoproteins in plasma. The microstructure of the eating behaviour was registered with a universal eating monitor, VIKTOR. Appetite was estimated by visual analogue rating scales and food intake by a 48-h recall. RESULTS: Serum leptin increased after 2 and 7 days of glucocorticoid administration (P < 0.01), and the food intake measured by VIKTOR after 7 days of treatment (P < 0.05). No statistically significant changes were however, found in the 48-h food- recall or in the subjective appetite registrations. Insulin levels were borderline elevated (P = 0.062) after treatment, but no significant changes of fasting glucose were seen. High density lipoprotein cholesterol (HDL) increased (P < 0.05), whilst low density lipoprotein cholesterol (LDL) decreased (P < 0.05). CONCLUSION: Food intake was elevated after glucocorticoid administration as observed with an objective, quantitative method, in spite of increased levels of circulating leptin. PMID- 12542566 TI - Paracetamol-induced hepatotoxicity at recommended dosage. AB - In patients who develop liver damage following moderate paracetamol overdose in the order of 5-10 g daily, recent fasting and nutritional impairment have been identified as key precipitants. Hepatotoxicity caused by paracetamol at recommended dosage, in the absence of exposure to enzyme-inducing drugs, has recently been described as an idiosyncratic phenomenon. The possible importance of fasting and malnutrition in this setting is uncertain. We report a severely malnourished 53-year-old woman who developed severe hepatotoxicity whilst receiving paracetamol at recommended dosage (4 g daily) following a period of fasting, in the absence of enzyme-inducing agents. Subsequent paracetamol exposure up to 2.6 g daily thrice weekly, in the setting of ongoing malnutrition and fasting as before, did not lead to recurrent liver damage. These findings indicate that paracetamol-related liver damage occurring within recommended dosage guidelines can be a dose-dependent rather than necessarily idiosyncratic phenomenon, at least in the setting of recent fasting and severe malnutrition. PMID- 12542565 TI - Smoking and relation to other risk factors in postmenopausal women with coronary artery disease, with particular reference to whole blood viscosity and beta-cell function. AB - OBJECTIVES: To investigate possible associations between smoking habits and other coronary risk factors in postmenopausal women with known coronary heart disease (CHD). SETTING: The study was conducted at a university clinic. SUBJECTS: A total of 118 postmenopausal women with CHD verified with angiography, consecutively recruited. INTERVENTIONS: Conventional treatment for CHD. The women were randomized to hormone replacement therapy (HRT) with transdermal 17-beta oestradiol and medroxyprogesterone acetate, or to a control group. RESULTS: Smokers were younger (P = 0.005), had lower body mass index (P = 0.04) and lipoprotein Lp(a) levels (P = 0.02) compared with nonsmokers. Smokers had reduced beta-cell function (homeostasis model assessment, P = 0.006), whereas whole blood viscosity (WBV) was higher at all shear rates. WBV was not affected by HRT over a 12-month period. Oestrone levels were higher in smokers. CONCLUSIONS: Smoking adversely affects insulin secretion (beta-cell function) and WBV in postmenopausal women with established CHD, which could be of importance as a mechanism for the increased risk of CHD in smokers. The importance of smoking as a risk factor, overrides the effect of Lp(a), which is lower in smokers compared with nonsmokers. PMID- 12542567 TI - Clinical relevance of isoproterenol challenge in tilt table testing in a patient with nonspecific repolarization abnormalities and exercise-induced syncope. PMID- 12542568 TI - Facial flushing and localized Castleman's disease: an extension of the POEMS syndrome phenotype. PMID- 12542570 TI - Diagnostic and prognostic role of cardiac troponins. PMID- 12542572 TI - Life satisfaction of people with intellectual disability living in community residences: perceptions of the residents, their parents and staff members. AB - BACKGROUND: Within the literature on quality of life (QoL), life satisfaction (LS) has emerged as a key variable by which to measure perceived well-being, which is referred to as subjective QoL. The LS self-reports of 93 residents with intellectual disability (ID) living in community-based residences were compared with reports about their LS completed by their staff and parents. METHOD: The residents were interviewed on their LS by social workers who did not belong to the staff of the interviewee's residence. The instrument used was the Life Satisfaction Scale (LSS). Staff and parents completed the short version of the LSS. RESULTS: Residents and staff's LS reports were positively correlated. However, significant differences were found between these two groups of informants when the residents were characterized as high functioning, had a low score in challenging behaviour, worked in an integrative employment setting and lived in an apartment. As opposed to staff/resident discrepancies, no differences were found between parents' and residents' LS reports. CONCLUSIONS: If residents cannot to be interviewed about their LS, then the parent is the preferred person to respond on behalf of the resident. The current study highlights the importance of including both objective measures (e.g. functional assessment characteristics) and subjective measures (e.g. LS) in order to get a better understanding of the QoL of people with ID. PMID- 12542573 TI - Motion analysis of stereotyped hand movements in Rett syndrome. AB - BACKGROUND: Rett syndrome is a genetic developmental disorder, and stereotyped hand movements are a striking feature of this condition. The aim of the present study was to subject these movements to objective analysis and compare the results in one girl at different ages. METHOD: The hand movements of a 10-year old girl with Rett syndrome were subjected to accurate, three-dimensional (3D) computerized motion analysis and compared to two-dimensional (2D) video analysis of the same girl at 3 years of age. RESULTS: Three-dimensional computerized analysis revealed regular patterns with strong coupling between the hands. Frequency analysis showed a dominant frequency at 1.2 Hz, with a higher component at 2.4 Hz that may relate to the activity of basic rhythm generators. The same coupling characteristics were extracted from standard, 2D video recordings made at the same time as the 3D capture. CONCLUSION: An informal video of the same girl taken when she was 3 years of age was analysed in the same way as the 2D video and showed the same characteristics, indicating the possible future use of automated video analysis for early screening and intervention evaluation. PMID- 12542574 TI - Magnetic resonance imaging, Down's syndrome and Alzheimer's disease: research and clinical implications. AB - BACKGROUND: The diagnosis of Alzheimer's disease (AD) remains at times difficult to make using available neuropsychological measures. Neuro-imaging is a relatively new form of detecting the changes associated with dementia. The present study investigated the role of magnetic resonance imaging (MRI) in diagnosing AD in adults with Down's syndrome (DS). METHODS: Subjects with DS and Alzheimer-type dementia were matched to non-demented controls with DS. Magnetic resonance imaging findings (i.e. volumetric and two-dimensional scans) were compared between the two groups in order to show a relationship between the changes of AD and structural MRI abnormalities. RESULTS: Specific structural abnormalities which are seen in non-intellectually disabled subjects with dementia are also found in individuals with both DS and AD. However, such findings cannot be used to diagnose clinical AD with good accuracy in adults with DS. A number of practical issues of patient compliance and over-sedation are demonstrated by the findings. CONCLUSIONS: Magnetic resonance imaging has an important but limited role to play in the management of AD in the population with DS. If intravenous sedation is used, medical support is essential to prevent a serious mishap. PMID- 12542575 TI - Parotid salivary immunoglobulins, recurrent respiratory tract infections and gingival health in institutionalized and non-institutionalized subjects with Down's syndrome. AB - BACKGROUND: The purpose of the present study was to evaluate the possible correlation between recurrent respiratory infections (RRIs), gingival health and the secretion rates of salivary immunoglobulins (Igs) in institutionalized (I) and non-institutionalized (NI) individuals with Down's syndrome (DS). METHODS: Stimulated parotid saliva was collected from nine I and 14 NI subjects with DS. Salivary flow rate, secretion rates of total salivary IgM, IgG and IgA, and the incidence of RRIs were determined. Gingival health was recorded by using the plaque index, the gingival index and the percentage of bleeding surfaces (BS). RESULTS: The mean salivary flow rate and parotid Ig secretion rates in the I group were 25% of those of the NI group. The prevalence of RRIs in the I group was more than twice that in the NI group. Oral hygiene and gingival health were significantly better in the NI group. CONCLUSIONS: The lower parotid Ig secretion rates in I individuals with DS might be a possible factor involved in their susceptibility to recurrent infections compared to NI subjects with DS. PMID- 12542576 TI - Prevalence, aetiology and comorbidity of severe and profound intellectual disability in Finland. AB - BACKGROUND: The aim of the present study was to describe the aetiology, associated impairments and prevalence of severe and profound intellectual disability (SPID) in Finland. METHODS: The number of people with SPID in the catchment area of the Paajarvi Centre for the Mentally Retarded, Lammi, Finland, (total population = 341,227) was calculated from the client register of this centre. Aetiological factors and background diagnoses for all subjects with SPID were analysed retrospectively. RESULTS: The number of people with SPID was 461, giving a prevalence of 0.13%. The aetiology of their SPID was genetic or congenital in 235 (50.9%) individuals, acquired in 89 (19.3%), genetic and/or acquired in 84 (18.3%), and unknown in 53 (11.5%) subjects. Out of the 53 individuals with an SPID of unknown origin, 48 (90.6%) had an associated impairment; the remaining five were the only members of the study group showing normal growth, and having neither dysmorphic features, physical abnormalities nor family members with ID. Out of the 461 subjects, 422 (91.5%) had between one and six associated impairments (total = 954), and the remaining 39 (8.5%) had SPID as their only impairment. Uncomplicated SPID was mainly of genetic or congenital origin, whereas all subjects with acquired encephalopathy had multiple disabilities. Speech defects, epilepsy and cerebral palsy were the most common associated impairments. CONCLUSIONS: Severe and profound ID almost always occurs concomitantly with other severe neurological or psychiatric impairments. The proportion of people with SPID described in the present study is similar to that found in Finland in 1966. The aetiology of SPID in the vast majority of cases is biopathological. PMID- 12542577 TI - Early play behaviour in children with 5p- (Cri-du-Chat) syndrome. AB - BACKGROUND: 5p- (Cri-du-Chat) syndrome (5p-S) is a well defined chromosomal condition. While the physical symptoms have frequently been documented, the developmental and behavioural aspects of the syndrome have not been explored adequately, especially in young children. METHOD: Mental level, and complexity and style of play were analysed in 10 children who were homogeneously selected with respect to their chronological age (range = 2-7 years). RESULTS: A high rate of distractability and a low level of object-directed behaviours were observed in the play sessions. The findings were compared to two comparison groups (subjects with Down's syndrome or Cornelia-de-Lange syndrome) matched for mental age in order to analyse the specificity of these behavioural features for young children with 5p-S. CONCLUSIONS: A low level of object-directed behaviours may be an early precursor of hyperactivity, distractability and stereotypy, which have been reported to be the characteristic features of the behavioural phenotype of older individuals with 5p-S. PMID- 12542578 TI - Mastery motivation in young children with Down's syndrome: relations with cognitive and adaptive competence. AB - BACKGROUND: Prior research on relations between motivation and competence have focused on cognitive competence, despite theoretical predictions that mastery motivation spurs behaviour that is effective in meeting the demands of one's environment, i.e. adaptive competence. Issues of adaptive competence are especially relevant for children with developmental delay since functional independence is an important long-term goal for these children. METHODS: In the present study, mastery motivation was examined in relation to both cognitive and adaptive competence in 5-year-old children with Down's syndrome (n = 41). RESULTS: Scores on mastery task and parent-report measures of mastery motivation were generally low, but positively related to scores on standardized measures of cognitive competence and adaptive competence. CONCLUSIONS: This is the first study of motivation-competence relations in pre-school-age children. The findings have implications for both developmental theory and early intervention efforts, and the authors hope that they will serve as a stimulus for future research on cross-domain relationships in atypical populations. PMID- 12542579 TI - Medical needs of people with intellectual disability require regular reassessment, and the provision of client- and carer-held reports. AB - BACKGROUND: Previous work has indicated a wide range of unmet medical health needs in people with intellectual disability (ID). METHODS: A profile of recorded medical needs was produced for 589 people with ID through a detailed search of individual medical and nursing case records. Specialist optometric and audiological assessments were offered, and reports were provided in technical and plain English terms. A Health Watch project delivered folders with copies of the plain English reports to 60 individuals and carers. RESULTS: The case record review indicated a wide range of medical disorders; however, exact diagnoses and counselling regarding underlying neurological conditions were seldom recorded. Assessed levels of hearing and vision loss were much greater than had been previously recognized. The Health Watch reports were welcomed by the clients and carers. CONCLUSIONS: If satisfactory healthcare is to be achieved for people with ID, medical needs must be monitored, regular specialist reassessments offered, access to specialist services facilitated and reports clearly explained to carers. PMID- 12542580 TI - Different presentations of late-detected phenylketonuria in two brothers with the same R408W/R111X genotype in the PAH gene. AB - Although the clinical heterogeneity of phenylketonuria (PKU) is well established, some questions about this condition remain. Subjects from the same family who share the same mutations in the phenylalanine hydroxylase (PAH) gene are expected to display similar disease courses, and therefore, when blood phenylalanine (Phe) levels, genotype and dietary treatment are all similar, differences in patient outcomes require additional explanations. The present authors describe two entirely different courses of late-detected PKU in two brothers with the same R408W/R111X genotype in the PAH gene. The older sibling was diagnosed with PKU at the age of 4 years and given treatment. His IQ was 97 at 26 years of age and moderate involvement of periventricular white matter was detected. The younger brother was diagnosed with PKU at the age of 11 months and given treatment. His IQ was < 25 at 22 years of age and severe dysmyelination changes were found by magnetic resonance imaging. The differences in the courses of the disease between these two brothers appear to be related to variations in their blood-brain barriers. PMID- 12542581 TI - Inpatient transfers to the intensive care unit: delays are associated with increased mortality and morbidity. AB - OBJECTIVE: To examine if delayed transfer to the intensive care unit (ICU) after physiologic deterioration is associated with increased morbidity and mortality. DESIGN: Inception cohort. SETTING: Community hospital in Ogden, Utah. PATIENTS: Ninety-one consecutive inpatients with noncardiac diagnoses at the time of emergent transfer to the ICU. We determined the time when each patient first met any of 11 pre-specified physiologic criteria. We classified patients as "slow transfer" when patients met a physiologic criterion 4 or more hours before transfer to the ICU. Patients were followed until discharge. INTERVENTIONS: None. MEASUREMENTS: In-hospital mortality, functional status at hospital discharge, hospital resources. MAIN RESULTS: At the time when the first physiologic criterion was met on the ward, slow- and rapid-transfer patients were similar in terms of age, gender, diagnosis, number of days in hospital prior to ICU transfer, prehospital functional status, and APACHE II scores. By the time slow transfer patients were admitted to the ICU, they had significantly higher APACHE II scores (21.7 vs 16.2; P =.002) and were more likely to die in-hospital (41% vs 11%; relative risk [RR], 3.5; 95% confidence interval [95% CI], 1.4 to 9.5). Slow transfer patients were less likely to have had their physician notified of deterioration within 2 hours of meeting physiologic criteria (59% vs 31%; P =.001) and less likely to have had a bedside physician evaluation within the first 3 hours after meeting criteria (23% vs 83%; P =.001). CONCLUSIONS: Slow transfer to the ICU of physiologically defined high-risk hospitalized patients was associated with increased risk of death. Slow response to physiologic deterioration may explain these findings. PMID- 12542582 TI - Mass media release of medical research results: an analysis of antihypertensive drug prescribing in the aftermath of the calcium channel blocker scare of March 1995. AB - BACKGROUND: Disclosure of medical research results to the public creates tension between lay medical reporters and the medical profession. OBJECTIVE: To explore the early effect of media attention on the risks associated with short-acting calcium channel blockers (CCBs) for treating hypertension after publication at a national meeting and following publication. DESIGN: Time-series analysis of prescription claims data. SETTING AND DATA SOURCE: National third-party pharmaceutical benefits manager. PATIENTS: Employed or retired persons and their families, 18 years of age or older, receiving prescription benefits from 1 of 4 national companies that contracted with the pharmaceutical benefits manager exclusively for prescription drug coverage. MEASUREMENTS: Prescription claims for antihypertensive drugs by fill date converted to a percentage of all cardiovascular drug claims. Data were grouped into weekly intervals before and immediately after the national release of negative information about CCBs on March 10, 1995 and following publication of the results on August 23, 1995. RESULTS: The most prevalent antihypertensive drugs were diuretics (21% of cardiovascular prescription claims) and calcium channel blockers (19%). A 10% decline in prescriptions filled for CCBs occurred 4 weeks following the intense media attention. Only prescriptions for long-acting calcium channel blockers declined. Alpha-1-blocker prescriptions increased by approximately the same amount that prescriptions for CCBs declined, suggesting substitution of one drug for the other. Changes in diuretic or beta-blocker prescriptions filled were not statistically significant. No immediate change in other cardiovascular drug classes occurred following journal publication. CONCLUSIONS: Intense media publicity regarding a controversial study measurably and unpredictably changed prescription claims. PMID- 12542583 TI - Primary care delivery is associated with greater physician experience and improved survival among persons with AIDS. AB - OBJECTIVE: It has been shown that greater physician experience in the care of persons with AIDS prolongs survival, but how more experienced primary care physicians achieve better outcomes is not known. DESIGN/SETTING/PATIENTS: Retrospective cohort study of HIV-infected patients enrolled in a large staff model health maintenance organization from 1990 through 1999. MEASUREMENTS: Adjusted odds of medical service delivery and adjusted hazard ratio of death by physician experience level (least, moderate, most) and service utilization. MAIN RESULTS: Primary care delivery by physicians with greater AIDS experience was associated with improved survival. After controlling for disease severity, patients cared for by the most experienced physicians were twice as likely to receive a primary care visit in a given month compared with patients of the least and moderately experienced physicians (P <.01). Patients of the least experienced physicians received the lowest level of outpatient pharmacy and laboratory services (P <.001) and were half as likely to have a specialty care visit compared with patients of the most and moderately experienced physicians (P <.05). Patients who received infrequent primary care visits by the least experienced physicians were 15.3 times more likely to die than patients of the most experienced physicians (P =.02). There was a significant increase in primary care services delivered to the population of HIV-infected patients receiving care in 1999, when highly active antiretroviral therapy (HAART) was in general use, compared with the time period prior to the introduction of HAART. CONCLUSIONS: Primary care delivery by physicians with greater HIV experience contributes to improved patient outcomes. PMID- 12542584 TI - Association of social stress, illicit drug use, and health beliefs with nonadherence to antiretroviral therapy. AB - OBJECTIVE: To assess the roles of socioeconomic status, social stability, social stress, health beliefs, and illicit drug use with nonadherence to antiretroviral therapy. DESIGN: Cross-sectional study. SETTING: Urban hospital clinic. PARTICIPANTS: One hundred ninety-six consecutive HIV-infected patients taking at least 1 antiretroviral medication, awaiting a visit with their primary care provider. METHODS: Patients were interviewed while waiting for a clinic appointment and were asked to fill out a 4-part survey with questions regarding antiretroviral adherence, illicit drug use, health beliefs, and social situation. Adherence was defined as the percentage of doses taken, i.e., the number of doses taken divided by the number of doses prescribed over a 2-week interval. Univariate and multivariate logistic regressions were performed to identify factors associated with nonadherence in different patient subgroups. MAIN RESULTS: Nonadherence to antiretroviral therapy was associated with active illicit drug use (adjusted odds ratio [AOR], 2.31; 95% confidence interval [95% CI], 1.17 to 4.58), eating fewer than 2 meals per day (AOR, 3.31; 95% CI, 1.11 to 9.92), and feeling as though pressures outside of the clinic affected patient's ability to take antiretroviral medications as prescribed (AOR, 2.22; 95% CI, 0.99 to 4.97). In patients with a history of injection drug use, nonadherence to antiretroviral therapy was independently associated with eating fewer than 2 meals per day (AOR, 17.54; 95% CI, 1.92 to 160.4) and active illicit drug use (AOR, 4.18; 95% CI, 1.68 to 10.75). In patients without any injection drug use, nonadherence was only associated with feeling as though pressures outside of clinic affected patient's ability to take antiretroviral medications as prescribed (AOR, 3.55; 95% CI, 1.07 to 11.76). Male-to-male sexual contact was associated with lower nonadherence in patients with an HIV risk factor other than injection drug use (AOR, 0.35; 95% CI, 0.13 to 0.95). A history of drug use but no illicit drug use within 6 months of the interview was not associated with an increased rate of nonadherence. CONCLUSIONS: Although our sample size was limited and variables that are not significant in subgroup analysis may still be associated with adherence, our results suggest that correlates of nonadherence are HIV risk factor specific. Strategies to increase antiretroviral adherence in HIV-infected patients should include social support interventions targeted at different risk factors for different patient groups. PMID- 12542585 TI - Evaluating medical students' skills in obtaining informed consent for HIV testing. AB - OBJECTIVE: To evaluate fourth-year medical students' abilities to obtain informed consent or refusal for HIV testing through a performance-based evaluation method. DESIGN: Student competence was assessed in a standardized patient interaction in which the student obtained informed consent or refusal for HIV testing. A previously validated 16-item checklist was completed by the standardized patient. A subset was independently reviewed and scored by a faculty member to calculate interrater reliability for this report. Student feedback on the assessment was elicited. SETTING: School of Medicine at the University of New Mexico. PATIENTS/PARTICIPANTS: All senior medical students in the class of 2000 were included. INTERVENTIONS: A 10-minute standardized patient interaction was administered within the context of a formal comprehensive performance assessment. MEASUREMENTS AND MAIN RESULTS: Seventy-nine students participated, and most (96%) demonstrated competence on the station. For the 15 specific items, the mean score was 25.5 out of 30 possible points (range, 13 to 30; SD, 3.5) on the checklist. A strong positive correlation (rs =.79) was found between the total score on the 15 Likert-scaled items and the score in response to the global item, "I would return to this clinician" (mean, 3.5; SD, 1.0). Scores given by the standardized patients and the faculty rater were well correlated. The station was generally well received by students, many of whom were stimulated to pursue further learning. CONCLUSIONS: This method of assessing medical students' abilities to obtain informed consent or refusal for HIV testing can be translated to a variety of clinical settings. Such efforts may help in demonstrating competence in performing key ethics skills and may help ensure ethically sound clinical care for people at risk for HIV infection. PMID- 12542586 TI - Diabetes control improved when inner-city patients received graphic feedback about glycosylated hemoglobin levels. AB - OBJECTIVE: To develop and test an inexpensive visual tool to help patients with diabetes improve glycemic control. METHODS: A multidisciplinary team developed a 1-page form, the "Take-home Diabetes Record" (THDR), providing feedback to patients by displaying per cent glycosylated hemoglobin (GHb) values graphically over time, with target levels highlighted. Patients with type 2 diabetes in an inner-city clinic were randomized to THDR use (n = 57) or not (n = 70) over 15 months. Self-care activities were discussed, linked with GHb results, and charted at each clinic visit. Initial and final GHb were compared. RESULTS: Mean GHb fell significantly in THDR patients (-0.94, P =.003), but not in control patients ( 0.18, P =.36). Mean GHb decrease was greater in THDR patients (P =.047). A greater proportion of THDR patients (51%) than control patients (18%) achieved a decrease in GHb >/=0.9 (P =.001). CONCLUSIONS: A graph linking GHb and self-care activities shows promise for improving glycemic control. PMID- 12542587 TI - Effect of violence on utilization of services and access to care in persons with HIV. AB - We analyzed the HIV Costs and Service Utilization Study data to determine the association of violence, assessed at baseline, with utilization of and access to health care at follow-up, among gay/bisexual male, heterosexual female, and heterosexual male HIV/AIDS patients. In multivariate analyses, male gay/bisexual violence victims had increased odds of reporting emergency department visits (odds ratio [OR], 1.74; 95% confidence interval [95% CI], 1.20 to 2.52), going without needed medical care because of expense (OR, 1.83; 95% CI, 1.06 to 3.18), and having poor ability to access medical specialists (OR, 1.96; 95% CI, 1.05 to 3.67). Further research is required to understand the association of violence with health care among gay/bisexual men with HIV/AIDS. PMID- 12542588 TI - Is the distal hyperplastic polyp a marker for proximal neoplasia? AB - CONTEXT: The current literature is unclear about the association between distal hyperplastic polyps and synchronous neoplasia (adenomatous polyps and cancer) in the proximal colon. OBJECTIVE: To estimate the prevalence of proximal neoplasia associated with distal hyperplastic polyps. DATA SOURCES: Database searches (medline and embase from 1966 to 2001) and manual search of the bibliographies of included and excluded studies, case reports, editorials, review articles, and textbooks of Gastroenterology. STUDY SELECTION: Studies describing the prevalence of proximal neoplasia in persons with distal hyperplastic polyps. DATA EXTRACTION: Demographics, clinical variables, study design, and prevalence of proximal neoplasia associated with various distal colorectal findings. DATA SYNTHESIS: Of 18 included studies, 12 involved asymptomatic individuals in which the pooled absolute risk of any proximal neoplasia associated with distal hyperplastic polyps was 25% (95% confidence interval [95% CI], 21% to 29%). In 4 studies where colonoscopy was performed irrespective of distal findings, the absolute risk was 21% (95% CI, 14% to 28%). The relative risk of finding any proximal neoplasia in persons with distal hyperplastic polyps was 1.3 (95% CI, 0.9 to 1.8) compared to those with no distal polyps. Among 6 studies of patients with symptoms or risk factors for neoplasia, the absolute risk of proximal neoplasia was 35% (95% CI, 32% to 39%) in persons with distal hyperplastic polyps. In 2 studies of screening colonoscopy, advanced proximal neoplasia (cancer, or a polyp with villous histology or severe dysplasia, or a tubular adenoma >/=1 cm) was present in 4% to 5% of persons with distal hyperplastic polyps, which was 1.5 to 2.6 times greater than in those with no distal polyps. CONCLUSIONS: In asymptomatic persons, a distal hyperplastic polyp is associated with a 21% to 25% risk for any proximal neoplasia and a 4% to 5% risk of advanced proximal neoplasia, and may justify examination of the proximal colon. Further study is needed to determine the risk of advanced proximal neoplasia associated with size and number of distal hyperplastic polyps. PMID- 12542589 TI - Helping journalists get it right: a physicians's guide to improving health care reporting. AB - News reports are the way that most people, including many physicians and scientists, first learn about new developments in medicine. Because these reports can raise awareness, influence behavior, and confer credibility, physicians should share responsibility with the media for accurate reporting. Physicians can work with reporters to avoid sensationalizing tentative findings, overstating benefits, and making inappropriate generalizations. This article includes pragmatic suggestions for crafting effective news releases and explaining numerical data. It details "rules of the road" for interviews. Working collaboratively with news reporters to improve the quality of medical stories in the lay press benefits patients and physicians alike. PMID- 12542590 TI - Racial and ethnic disparities in the use of health services: bias, preferences, or poor communication? AB - African Americans and Latinos use services that require a doctor's order at lower rates than do whites. Racial bias and patient preferences contribute to disparities, but their effects appear small. Communication during the medical interaction plays a central role in decision making about subsequent interventions and health behaviors. Research has shown that doctors have poorer communication with minority patients than with others, but problems in doctor patient communication have received little attention as a potential cause, a remediable one, of health disparities. We evaluate the evidence that poor communication is a cause of disparities and propose some remedies drawn from the communication sciences. PMID- 12542591 TI - On the prevention and treatment of exaggeration. PMID- 12542592 TI - Delays in transfer to the ICU: a preventable adverse advent? PMID- 12542593 TI - Quality primary care for HIV/AIDS: how much HIV/AIDS experience is enough? PMID- 12542594 TI - Hepatocellular carcinoma in sustained responders of interferon-treated chronic hepatitis C. PMID- 12542595 TI - Non-alcoholic steatohepatitis: what is it, and why is it important in the Asia Pacific region? AB - Non-alcoholic steatohepatitis (NASH), is a critical link in the chain of metabolic fatty liver disorders that spans steatosis to cryptogenic cirrhosis. It is the hepatic manifestation of the insulin resistance (or metabolic) syndrome, and provides a clue to understanding fibrotic progression of other chronic liver diseases, particularly hepatitis C. Non-alcoholic steatohepatitis is often the first clinical indication of insulin resistance, with its complications of high blood pressure, coronary heart disease and type 2 diabetes. Among those with risk factors, NASH is common: present in at least 20% of obese adults or children with or without type 2 diabetes, and at least 5% of those overweight. With emerging urbanization, increasing affluence and behavioral changes of physical inactivity and high fat/energy-excessive diet, type 2 diabetes has become common in Asia and the western Pacific rim. The rates range from 7-40%, which in countries like Japan represents a 3-20-fold increase (depending on age) over the last 20 years. The increase is associated with central adiposity, insulin resistance, hepatic steatosis and NASH. After cancer, cirrhosis from NASH is now the second most common age-related cause of death in type 2 diabetes. Reversing these trends must become a public health priority; the first awakenings were evident in Taiwan at the time of this meeting. In order to stimulate clinicians to think more about the importance of metabolic liver disease for development of cirrhosis, this review will cover clinical and laboratory features, natural history and an approach to diagnosis and management of NASH. Some emerging concepts on pathogenesis will be mentioned briefly, but the emphasis will be on the potency of lifestyle adjustments (physical activity and diet) to prevent or reverse fatty liver disorders. PMID- 12542596 TI - Is smoking an indirect risk factor for the development of ulcerative colitis? An age- and sex-matched case-control study. AB - BACKGROUND: It has been suggested that smoking protects against the development of ulcerative colitis (UC). Evidence is mainly driven from the way data from a multitude of case-control studies have been interpreted. METHODS: An age- and sex matched case-control study was conducted to further assess the association between history of smoking, past surgery, childhood, and other potential causative factors with the development of UC using the answers to a detailed questionnaire. The data were analyzed using univariate analysis and logistic regression. The results are presented as odds ratios (OR) and 95% confidence intervals. RESULTS: One hundred and two cases and an equal number of matched controls were included in the study. Using a three-level comparison, at the age of diagnosis, the risks of developing UC were 0.41 (0.19-0.87), 3.45 (1.62-7.35) and 0.78 (0.44-1.37) for smokers, ex-smokers and non-smokers, respectively. When compared to individuals who have never smoked, ex-smokers were at a higher risk of developing the disease (OR = 3.00 (1.38-6.51)). The specific history of quitting smoking prior to the age of onset of symptoms was associated with an increased risk for developing the disease (OR = 3.45 (1.62-7.35)). CONCLUSIONS: Active smoking was associated with a low risk for the development of UC, but the lack of history of smoking was not associated with an increase in the risk. History of quitting smoking prior to the onset of symptoms, in contrast, was associated with a significant increase in the risk of developing the disease. These findings make the theory of a simple protective effect of smoking on the development of UC difficult to justify. It may be plausible to suggest that the withdrawal of the immunosuppressive effect of smoking triggers the disease onset in a genetically susceptible individual or simply unmasks its symptoms. PMID- 12542597 TI - c-kit gene mutation at exon 17 or 13 is very rare in sporadic gastrointestinal stromal tumors. AB - BACKGROUND AND AIM: Gastrointestinal stromal tumors (GIST) are the most common mesenchymal tumors of the human gut. They frequently have gain-of-function mutations of the c-kit gene, which encodes a receptor, tyrosine kinase. The mutations were found at exon 11 in most cases, and either at exon 9 or at exon 13 in rare cases. Recently, we found a family with multiple GIST and a gain-of function mutation at exon 17. The family was the first reported GIST case with c kit gene mutation at exon 17 including sporadic GIST. Although we previously reported that the c-kit gene mutation at exon 17 was not detected in 124 sporadic GIST by single-strand conformation polymorphism (SSCP) analysis, the mutation at exon 17 observed in the familial GIST was detectable by the use of direct sequencing but not by our SSCP method. In the present study, we examined the mutations at exon 17 and exon 13 by using direct sequencing. METHODS: Genomic DNA was extracted from formalin-fixed, paraffin-embedded GIST tissues. We could obtain 143 sporadic GIST cases appropriate for DNA analysis at exon 17 and 141 at exon 13. Exons 17 and 13 were amplified by using polymerase chain reaction and direct sequencing was conducted. RESULTS: No mutation was found at exon 17, and only one case with the mutation at exon 13 was observed. The GIST with the mutation at exon 13 was large and showed frequent mitosis, and the patient died of the recurrent GIST 3 years after the first operation. CONCLUSION: The mutation at exons 17 or 13 was considered to be very rare in sporadic GIST. PMID- 12542598 TI - Beneficial effects of terlipressin in hepatorenal syndrome: a prospective, randomized placebo-controlled clinical trial. AB - BACKGROUND AND AIM: Hepatorenal syndrome (HRS) occurs in about 18% of cirrhotic patients with ascites and is characterized by intense renal vasoconstriction, low glomerular filtration rate and preserved tubular function and normal renal histology. The aim of the present study was to examine the effects of terlipressin on renal function, systemic hemodynamics and clinical outcome in patients with HRS. METHODS: The study was a randomized controlled single-blind trial. We randomly assigned 24 consecutive patients with HRS to treatment with terlipressin 1 mg i.v. at 12-h intervals (group A; n = 12) or placebo at 12-h intervals (group B; n = 12). The end-point of the study was improvement in renal functions defined as reversal of HRS and survival at 15 days. RESULTS: The two groups were comparable at baseline. After treatment with terlipressin, urine output significantly (P < 0.05) increased progressively in group A (day 4, 960 +/ 40 mL/24 h; day 8, 1068 +/- 56 mL/24 h) compared with group B (day 4, 451 +/- 40 mL/24 h; day 8, 291 +/- 45 mL/24 h). Creatinine clearance improved (P < 0.05) in group A (day 4, 20.2 +/- 2.1 mL/min; day 8, 35 +/- 2.8 mL/min) compared with group B (day 4, 11.3 +/- 1.3 mL/min; day 8, 9.3 +/- 1.7 mL/min). Serum creatinine decreased in group A but not in group B (day 8, 1.6 +/- 0.01 compared with 3.9 +/ 0.26, P < 0.05). Mean arterial pressure increased significantly (P < 0.05) in group A. Terlipressin administration was associated with transient self-limiting side-effects including crampy abdominal pain in two patients and cardiac arrhythmias in three patients. Five of the 12 patients survived in group A compared with none in group B at day 15 (P < 0.05) and all survivors had reversal of HRS. CONCLUSION: In patients with HRS, terlipressin significantly improved renal functions and systemic hemodynamics, and showed a trend towards better clinical outcome. The drug merits further evaluation with different dosages and longer schedules. PMID- 12542599 TI - Impaired gallbladder mucosal function in aged gallstone patients suppresses gallstone recurrence after successful extracorporeal shockwave lithotripsy. AB - BACKGROUND: Absorption of water, as well as emptying of bile, are important functions of the gallbladder. We studied the changes of gallbladder function with age in gallstone patients and their influence on the outcome of extracorporeal shockwave lithotripsy (ESWL). METHODS: (i) A total of 123 consecutive patients with complete stone clearance by ESWL were examined. Gallbladder emptying was assessed before treatment using intravenous cholecystography. After stone clearance, the recurrence of gallstones was monitored by using ultrasonography. Cox regression analysis was used to determine the risk factors associated with stone recurrence. (ii) Gallbladder bile was sampled from 59 gallstone patients during surgery. Biliary cholesterol, phospholipids, and total bile acids were simultaneously quantified by using gas-liquid chromatography. RESULTS: Impaired gallbladder function, but not gallstone recurrence, was more frequently observed in older patients (>/=65 years old) than in younger patients (<65 years old). Cox regression analysis revealed that poor gallbladder emptying was an independent predictor of stone recurrence after ESWL in the total study population, but not in the older patients (>/=65 years old). Analysis of bile from surgically treated patients with cholesterol stones showed a significantly higher total lipid concentration and a shorter nucleation time in the younger group (<65 years old), but the cholesterol saturation index did not differ between the younger and older groups. CONCLUSIONS: Our data suggest that the reduced concentrating function of the gallbladder in elderly gallstone patients helps to counteract stone recurrence despite their abnormal gallbladder motility. Therefore, aged gallstone patients may be preferentially treated by a non-surgical strategy. PMID- 12542600 TI - Long-term follow up of gastric low-grade mucosa-associated lymphoid tissue lymphoma by endosonography emphasizing the application of a miniature ultrasound probe. AB - BACKGROUND AND AIMS: Endoscopic ultrasonography (EUS) is a useful tool for the evaluation of gastric wall infiltration including gastric lymphoma. The aims of this study were to characterize gastric low-grade mucosa-associated lymphoid tissue (MALT) lymphoma according to EUS findings and to evaluate the role of a miniature ultrasound probe in the long-term follow up. METHODS: From January 1994 to March 2002, 20 patients were proven to have gastric low-grade MALT lymphoma. Endoscopic ultrasonography was performed with a conventional echoprobe and/or a miniature ultrasound probe for initial staging and a miniature ultrasound probe was performed during follow up. All patients positive for Helicobacter pylori received a 2-week course of omeprazole, amoxicillin and clarithromycin. RESULTS: Helicobacter pylori infection was found in 17 (85%) patients. In all patients, H. pylori was eradicated after treatment. Initial EUS showed significantly greater wall thickness (6.1 +/- 3.0 mm) in MALT lymphoma patients when compared with control (2.8 +/- 0.3 mm). The infiltrative patterns included wall thickening (3.5 14.1 mm) in 18 patients: stage E-I1 in 16 (mucosa and/or submucosa), stage E-I2 in one and stage E-II in one. Complete regression of MALT lymphoma following treatment for H. pylori was noted in 14 patients, with a mean duration of 11.3 +/ 9.1 months. Follow-up miniature ultrasound probe sonography showed comparative reduction in wall thickness (P < 0.05). CONCLUSIONS: Endoscopic ultrasonography plays a valuable role in the initial staging and long-term follow up of gastric low-grade MALT lymphoma. The application of a miniature ultrasound probe enables adequate evaluation in the majority of these patients, with additional benefits. PMID- 12542601 TI - Influence of lansoprazole, famotidine, roxatidine and rebamipide administration on the urea breath test for the diagnosis of Helicobacter pylori infection. AB - BACKGROUND AND AIM: The sensitivity of the urea breath test (UBT) has been reported to be influenced by the administration of omeprazole, lansoprazole and ranitidine. However, it is unclear whether other H2 receptor antagonists (H2RA), except ranitidine, and rebamipide, a mucosal protective agent, affect UBT sensitivity. The aim of this study is to clarify the effects of lansoprazole, famotidine, roxatidine and rebamipide administration on UBT sensitivity. METHODS: Subjects comprised 30 volunteers with Helicobacter pylori infection. All subjects were examined by the 13C-UBT on four occasions: (i) without medication (control); (ii) after the administration of 30 mg lansoprazole (u.i.d) for 14 days; (iii) after the administration of 100 mg rebamipide (t.i.d) for 14 days; and (iv) after the administration of 20 mg famotidine or 75 mg roxatidine (b.i.d) for 14 days. In the H2RA study, individuals were randomized into two groups of 15 subjects and were administered either famotidine or roxatidine. RESULTS: Five of the 30 cases administered lansoprazole and one of the 15 cases given roxatidine gave a false negative UBT result. No negative UBT results were observed in patients administered famotidine or rebamipide. CONCLUSION: Patients showing negative UBT results during the administration of proton pump inhibitors and H2RA should be re examined after the cessation of these drugs to confirm the true negativity of H. pylori infection. PMID- 12542602 TI - Clinical characteristics of Japanese reflux esophagitis patients as determined by Los Angeles classification. AB - BACKGROUND: Recent studies have shown that the number of patients with reflux esophagitis is increasing in Japan, but the prevalence and risk factors associated with reflux esophagitis in Japanese patients are not well defined. METHODS: By using all endoscopic records in the Katta General Hospital from April through to September 1999, we identified 392 patients. We examined the Los Angeles classification, peptic ulcer, gastric mucosal atrophy, hiatal hernia and other medical variable factors for their contribution to esophagitis in the patients. RESULTS: Patients (13.8%) were diagnosed as having reflux esophagitis with a mucosal break. In a multivariate analysis, reflux esophagitis was associated with hiatal hernia (odds ratio (OR) 2.276, 95% confidence interval (CI) 1.164-4.450), with patients over 65 years of age (OR 2.521, 95% CI 1.238 5.134) and the open type of gastric mucosal atrophy (OR 0.420, 95% CI 0.225 0.785). There was no significant difference between esophagitis and Helicobacter pylori infection and peptic ulcer. CONCLUSIONS: We observed that age, hiatal hernia and a lower rate of gastric mucosal atrophy were associated with the proportion of mucosal breaks accompanying esophagitis. PMID- 12542603 TI - Lafutidine increases hepatic blood flow via potentiating the action of central thyrotropin-releasing hormone in rats. AB - BACKGROUND: Lafutidine, (+/-)-2-(furfurylsulfinyl)-N-[4-[4-(piperidinomethyl)-2 pyridyl]oxy-(Z)-2 butenyl]acetamide, is a newly synthesized histamine H2 receptor antagonist and possesses a cytoprotective efficacy, which comprises mucin biosynthesis and stimulation of gastric blood flow mediated through capsaicin sensitive sensory neurons and endogenous calcitonin gene-related peptide (CGRP). In the present study, an effect of lafutidine on hepatic blood flow was investigated in rats that received an intracisternal injection of a subthreshold dose of thyrotropin-releasing hormone (TRH) analog, RX 77368. METHODS: Change in hepatic blood flow was determined by laser Doppler flowmetry. Male Wistar rats were anesthetized with urethane (1.5 g/kg, i.p.), and positioned on a stereotaxic apparatus. An abdominal incision was made, and a probe of laser Doppler flowmeter was placed on the surface of the liver. After a 60-min stabilization, basal hepatic blood flow was measured for 30 min, and lafutidine (0.5, 1, 3, 5 or 10 mg/kg) or vehicle was injected into the portal vein and a subthreshold dose (1.5 ng) of RX 77368 was injected intracisternally. Hepatic blood flow was monitored for 120 min postinjection. To investigate a role of capsaicin-sensitive sensory neurons and endogenous CGRP, systemic capsaicin treatment (125 mg/kg, s.c., 10-14 days before) and intravenous infusion of a CGRP receptor antagonist, human CGRP (8-37) (15 micro g/kg as a bolus, followed by infusion at 3 micro g/kg/h) were performed, respectively. RESULTS: Intracisternal injection of RX 77368 (1.5 ng) or intraportal lafutidine (10 mg/kg) by itself did not affect hepatic blood flow, but co-injection of intracisternal RX 77368 (1.5 ng) and intraportal lafutidine (5 mg/kg) increased it with peak response at 30 min postinjection. The effect of lafutidine on hepatic blood flow in rats given RX 77368 was dose-related over the range 1-5 mg/kg. By contrast, intracisternal injection of RX 77368 (1.5 ng) did not change hepatic blood flow in rats injected with another histamine H2 receptor antagonist, famotidine (5 mg/kg), intraportally. The stimulatory effect of co injection of TRH analog and lafutidine was abolished by systemic capsaicin treatment and CGRP antagonist. CONCLUSION: These data suggest that lafutidine increases hepatic blood flow by sensitizing the liver to the action of central TRH via both capsaicin-sensitive sensory neurons and endogenous CGRP in urethane anesthetized rats. PMID- 12542604 TI - Clinical and laboratory differentiation of cirrhosis and extrahepatic portal venous obstruction in children. AB - BACKGROUND AND AIM: Differentiation between cirrhosis and extrahepatic portal venous obstruction (EHPVO) in children presenting with features of portal hypertension is important for cost-effective management and proper resource utilization. We undertook this study to differentiate clinical and laboratory features between these two groups of patients. METHODS: Clinical features and laboratory parameters at presentation of children with portal hypertension and cirrhosis of the liver were analyzed. The variables analyzed were age at presentation, duration of symptoms, incidence and frequency of upper gastrointestinal (GI) bleeding, encephalopathy, jaundice, ascites, splenomegaly, and presence of dilated abdominal veins. The laboratory parameters studied were hemoglobin, prothrombin time, serum bilirubin, albumin, aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase. Two groups were compared using appropriate statistical methods; and the differentiating features were analyzed using logistic regression analysis. RESULTS: Of the total of 120 cases, cirrhosis was diagnosed in 28.3% and EHPVO in 71.6%. Children with EHPVO, in comparison to cirrhosis, had significantly higher frequency of upper GI bleeding (61.6%vs 14.7%), increased number of previous bleeding episodes (2.7 +/- 0.5 vs 1.2 +/- 0.4), longer duration of symptoms (25.7 +/- 4.6 vs 12.3 +/- 3 months) and a lower frequency of jaundice (2%vs 76.4%). Low hemoglobin (6.4 +/- 2.7 g/dL) and preserved liver functions characterized by normal bilirubin, albumin levels and prothrombin time were observed in EHPVO cases. Cirrhosis patients had higher hemoglobin (8.8 +/- 2.8 g/dL) and abnormal liver function tests. CONCLUSION: Presence of UGI bleeding and the absence of jaundice are 97.5% accurate in predicting diagnosis of EHPVO. PMID- 12542605 TI - Anti-Helicobacter pylori therapy in India: differences in eradication efficiency associated with particular alleles of vacuolating cytotoxin (vacA) gene. AB - BACKGROUND AND AIMS: The efficiency of Helicobacter pylori eradication varies geographically, as do many parameters that might affect therapeutic efficiency, including bacterial genotype. The aim of the present study was to determine the efficiency of H. pylori eradication using a 10-day proton pump inhibitor-based triple-therapy regimen (omeprazole, clarithromycin and amoxycillin) in an eastern Indian patient population, and to find out the relationship, if any, of the success or failure of the therapy to known features of bacterial genotype. METHODS: Helicobacter pylori infections were analyzed in 66 duodenal ulcer patients by upper gastrointestinal endoscopy, rapid urease tests, histology and culture. The cytotoxin-associated gene (cagA) and vacuolating cytotoxin (vacA) gene status of cultured strains were studied by polymerase chain reaction. Treatment was given for 10 days and endoscopy was repeated at 4 and 12 weeks post therapy to monitor ulcer healing and H. pylori eradication. RESULTS: Ulcer healing was observed in 60 patients (96.77%). Helicobacter pylori was eradicated in 41 (62.12% intention to treat, 66.13% per protocol) of the 66 duodenal ulcer patients, but not in the other 25. The bacteria from 47 patients were genotyped. The only significant disease-associated difference in patterns observed was that the vacA m1 allele was represented more disproportionately among patients with eradication failures (68%) than in those with successful eradication (39%) (P < 0.05) No significant association of vacAs1 (signal sequence allele) or cag pathogenicity island status with persistence was detected. CONCLUSIONS: This study highlights the public health need for cheaper, more cost-effective anti-H. pylori therapies for developing countries, and suggests that subtle features of bacterial genotype can influence therapeutic efficiency. The possibility that particular vacA mid region alleles affect persistence, perhaps through toxin action on particular gastric cell types, merits further study. PMID- 12542606 TI - Meta-analysis of randomized controlled trials on the benefits and risks of using cisapride for the treatment of gastroesophageal reflux in children. AB - BACKGROUND AND AIM: Gastroesophageal reflux is a common problem in infancy. Cisapride is a commonly used therapy for gastroesophageal reflux in children. In view of recent concern regarding adverse effects this study aims to evaluate the benefits and risks of cisapride for the treatment of gastroesophageal reflux in children. METHODS: A meta-analysis of randomized controlled trials of cisapride using a random-effects model. RESULTS: Ten trials involving 415 children were identified. There was no evidence of a significant reduction in vomiting severity with cisapride as measured by a clinical score (five trials, standardized weighted mean difference -0.18; 95% confidence interval (CI) -0.51 to 0.15). Twenty-four-hour esophageal pH monitoring data showed the mean reflux index was significantly lower in the children treated with cisapride compared with controls (five trials, weighted mean difference -6.24; 95% CI -8.81 to -3.67). With cisapride treatment, there was no reduction in the mean number of reflux episodes lasting greater than 5 min (three trials, weighted mean difference -0.72; 95% CI 1.92 to 0.47) or in the number of children with esophagitis at final follow up compared with baseline (two trials, relative risk 0.80; 95% CI 0.40 to 1.61). There was no significant difference in reported side-effects or adverse events (six trials, relative risk 1.16; 95% CI 0.95 to 1.41). CONCLUSIONS: No clinically important benefits of cisapride in children with gastroesophageal reflux have been demonstrated. Nor was there any evidence of adverse or harmful events. PMID- 12542607 TI - Enhancement of aquaporin-3 by vasoactive intestinal polypeptide in a human colonic epithelial cell line. AB - BACKGROUND: Vasoactive intestinal polypeptide (VIP) plays an important role in water transport in the intestine. Several specialized channels termed aquaporins (AQP) facilitate water transport in the gastrointestinal tract. Aquaporin-3 localizes to epithelial cells in the human small intestine and colon. However, the regulatory mechanisms underlying the functions of AQP3 remain unclear. To characterize the regulation of AQP3 expression by VIP, we studied messenger (m)RNA expression, protein expression and DNA binding activity in a human colonic epithelial cell line, HT-29. METHOD: Human colonic epithelial cells, HT-29, were incubated with VIP (10-12-10-7 M). The cells were treated with protein kinase-A (PK-A) inhibitors (H-89, H-9) or chloride channel-blockers (diphenylamine-2 carboxylate (DPC), 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPD)). The expression of AQP3 mRNA and protein was determined by Northern blot and Western blot, respectively. The DNA-binding activities of cyclic adenosine monophosphate (cAMP) response elements/activating transcription factor (CRE/ATF)) in the nuclear extract were determined by electrophoretic mobility shift assay. RESULTS: Aquaporin-3 mRNA was up-regulated at a concentration of 10-10 M VIP. The expression of AQP3 protein was enhanced at 3 h after addition of VIP. The PK-A inhibitors (H-89, H-9) inhibited the expression of AQP3 mRNA enhanced by VIP and cAMP. The gel shift assay of CRE/ATF in HT-29 cells revealed a single band. CONCLUSION: These results indicate that VIP upregulated the expression of AQP3 mRNA and protein, and that a cAMP-dependent pathway mediated this effect in a human colonic epithelial cell line, HT-29. PMID- 12542608 TI - Low-dose ethanol attenuates gut ischemia/reperfusion-induced liver injury in rats via nitric oxide production. AB - BACKGROUND AND AIMS: The acute administration of low-dose ethanol was demonstrated to attenuate liver injury elicited by gut ischemia/reperfusion (I/R). Nitric oxide (NO) has been found to be a modulator of adhesive interactions between leukocytes, platelets, and endothelial cells, but there has been much controversy about the effects of ethanol on NO regulation. The objective of this study was to investigate the role of NO in ethanol-reduced hepatic microvascular dysfunction elicited by gut I/R. METHODS: Male Wistar rats were exposed to 30 min of gut ischemia followed by 60 min of reperfusion. Intravital microscopy was used to monitor leukocyte recruitment and non-perfused sinusoids (NPS). Plasma alanine aminotransferase (ALT) activities were measured 6 h after the onset of reperfusion. In another set of experiments, ethanol (10%, 1 g/kg) was administered before ischemia. RESULTS: Gut I/R elicited increases in the number of stationary leukocytes, NPS, and plasma ALT activities; all of which were attenuated by pretreatment with ethanol or an NO donor. Gut I/R caused the apoptosis of hepatocytes, which was prevented by pretreatment with ethanol. Pretreatment with an NO synthase inhibitor diminished the protective effects of ethanol. The administration of ethanol increased plasma nitrite/nitrate levels. CONCLUSION: These results suggest that low-dose ethanol attenuates the gut I/R induced hepatic microvascular dysfunction and sequential liver injury by increasing sinusoidal NO levels. PMID- 12542609 TI - Failure of therapeutic vaccination using hepatitis B surface antigen vaccine in the immunotolerant phase of children with chronic hepatitis B infection. AB - AIM: The aim of this study was to investigate the efficacy of specific hepatitis B virus (HBV) vaccination as active immunotherapy in treating chronic hepatitis B (CHB) infection during the immune-tolerant phase in children with normal aminotransferase levels and high viral load. METHODS: Fifty-one immunotolerant patients were randomly and prospectively recruited into two groups. Group 1 included 23 patients that were vaccinated with three standard injections of the GenHevac B vaccine in the deltoid or quadricep muscle, initially, and at 30 days and 60 days, for specific immunization. Group 2 contained 28 patients who did not receive any medication or vaccination and were recruited as the control group. Post-vaccination evaluation was performed at 6 months from the first injection and at the end of the 12th month by serological and virological analyses. A response criterion to therapy was defined as loss of HBV-DNA in serum and hepatitis B early antigen (HBeAg) seroconversion (loss of HBeAg, development of antibody to HBeAg (anti-HBe)). RESULTS: The mean alanine aminotransferase (ALT) value in group 1 at the beginning of the vaccination was 33.6 +/- 8.1 IU/L; this changed to 31.7 +/- 9.0 IU/L at 6 months after first injection and 29.2 +/- 7.1 IU/L at the end of 12 months (P > 0.05). In this group, mean HBV-DNA load at the starting point of the vaccination was 3,709 +/- 1,126 pg/mL; this value changed to 3,569 +/- 726 pg/mL at the sixth month and 3,295 +/- 832 pg/mL at the 12th month (P > 0.05). In group 2, the mean ALT values at the beginning of therapy, and at the 6th and 12th month were 32 +/- 8 IU/L, 31.8 +/- 8 IU/L, and 29.7 +/- 7 IU/L, respectively (P > 0.05), and the mean viral load of HBV-DNA values were 3,827 +/- 1,375 pg/mL, 3,498 +/- 886 pg/mL, and 3,059 +/- 731 pg/mL, respectively (P > 0.05). The load of HBV DNA of all patients in both groups was greater than 2,000 pg/mL. There was no statistically significant difference in the mean ALT values and mean viral load of HBV DNA (P > 0.05) between group 1 and group 2 at the end of the 6th and 12th months. Except for one each patient in each group, hepatitis B surface antigen (HBsAg) and HBeAg clearance or antibody to HBsAg (anti-HBs) and anti-HBe seroconversion were not observed during the follow-up period (P > 0.05). CONCLUSION: In this study, comparison of vaccinated and unvaccinated groups of immunotolerant children with CHB infection showed no difference in the clearance of HBV DNA and seroconversion of HBeAg to anti-HBe. Different immunization protocols should be considered for future investigations in the immunotolerant phase of children with CHB infection. PMID- 12542610 TI - Hepatobiliary and pancreatic: an old man with jaundice and abdominal pain. PMID- 12542611 TI - Gastrointestinal: iatrogenic gastrointestinal bleeding. PMID- 12542613 TI - Simultaneous occurrence of adenoma, focal nodular hyperplasia, and hemangioma of the liver: are they derived from a common origin? AB - The association between hepatic hemangioma (HH) and focal nodular hyperplasia (FNH) or the association between FNH and hepatic adenoma (HA) has been reported. The authors report a case in which FNH, HH, and HA simultaneously appear in the liver. A 25-year-old woman was admitted to the Department of Surgery of the University of Catania (Italy), after presenting pain in the right hypocondrium. No therapy with oral contraceptives, no pregnancy and no abnormalities of the laboratory tests were found. Ultrasonography and computed tomography scans revealed four masses with the characteristics of HH, HA, FNH, and a hydatid cyst located, respectively, in segments II, IV, IV, and V of the liver. The surgical procedures performed were hemangioma and adenoma enucleation and en bloc resection of the FNH, hydatid cyst and gallbladder. No complications were recorded in the postoperative period and the patient was discharged from the hospital after 1 week. A pathological examination confirmed the preoperative diagnosis. To the best of our knowledge, the association of HH, FNH, and HA has never been reported. A common pathogenesis has clearly been demonstrated for hepatocytes and other cell types. The simultaneous presence of these three different kinds of tumor suggest that HH, FNH and HA could be the different expression of the same malformative anomaly. PMID- 12542614 TI - Octreotide for bleeding as a result of hepatocellular carcinoma invasion of the gastrointestinal tract. PMID- 12542617 TI - Conventional versus resin-modified glass-ionomer cement for Class II restorations in primary molars. A 3-year clinical study. AB - OBJECTIVE: To compare the clinical performance of two glass-ionomer cements (GICs)for Class II restorations in primary molars: a conventional cement (Fuji II) and a resin-modified cement (Vitremer). DESIGN: Split mouth and random assignment to the two materials were used for the majority of the molars. SAMPLE AND METHODS: Forty consecutive 4-7-year-old children were included. One operator made 115 restorations: 53 with Vitremer and 62 with Fuji II. The restorations were evaluated clinically, radiographically and from colour photographs. RESULTS: The cumulative success rate of the Vitremer restorations was 94% and that of the Fuji II restorations 81%. The difference is statistically significant. The risk of a failed restoration was more than five times higher with Fuji II than with Vitremer as the restorative material. Of the 13 unsuccessful restorations, seven had lost their retention, four had secondary caries, and two were fractured. CONCLUSIONS: The resin-modified GIC offered advantages over the conventional GIC for restoring approximal caries in primary molars. PMID- 12542618 TI - Self-report treatment techniques used by dentists to treat dentally anxious children: a preliminary investigation. AB - OBJECTIVE: To investigate the extent to which dentists report their encouragement to dentally anxious children to blunt (distract from the procedure) or monitor (give information about the procedure) during treatment. SETTING: British Society of Paediatric Dentistry conference. SUBJECTS AND METHODS: One hundred and two of 155 delegates (66% response rate) completed a Monitor Blunter Treatment Checklist (MBTC), which was included in their conference pack. Participants were asked to tick all of the 10 statements (five monitoring and five blunting) that reflected how, in general, they would treat a dentally anxious child. RESULTS: Dentists chose significantly more monitoring than blunting statements. Treatment strategies relating to explaining procedures were endorsed by the majority of dentists, while distraction was chosen by less than a third. CONCLUSION: Dentists tend to use monitoring strategies that adhere generally to the principles of the Tell-Show-Do technique. This may have implications if the anxious child has a 'blunting' coping style. PMID- 12542619 TI - Knowledge of physicians in hospital emergency rooms in Israel on their role in cases of avulsion of permanent incisors. AB - OBJECTIVE: The purpose of this study was to assess the knowledge of physicians in hospital emergency room care on their possible role in cases of traumatic loss of permanent incisors. METHODS: A questionnaire was sent to directors of emergency room facilities in hospitals in Israel regarding the medical team at the emergency room in the hospital, availability of an in-house dental service and existence of a protocol for management of avulsed teeth. Another questionnaire, completed by physicians in the emergency room, contained demographic questions and questions regarding their knowledge of management of avulsed permanent teeth. RESULTS: The findings of the study showed that only 4% (12 of 335 physicians) would provide an appropriate initial treatment that, if followed by treatment by a dentist as recommended, could save the avulsed tooth. No correlation was found between knowledge of appropriate treatment of avulsed teeth and type of specialty, years of experience, previous exposure to information on trauma to the teeth and gender. The only significant association was found between physicians' knowledge and having a dentist spouse. Most of the physicians had never attended any formal seminar or lecture or received printed instructions on the management of avulsed teeth. CONCLUSIONS: Medical students and physicians in hospitals' emergency rooms should be made aware of their possible role in cases of avulsion of permanent teeth, in order to minimize late complications associated with such injuries. One possible way to achieve this goal is through education during and after training and introduction of a formal protocol for treatment of avulsed permanent teeth and other dental injuries. PMID- 12542620 TI - The effect of sociodemographic factors on dental anxiety in children seen in a suburban Nigerian hospital. AB - OBJECTIVE: This paper attempts to determine the sociodemographic factors that relate to dental anxiety in suburban African children. METHODS: A short form of the Dental subscale of the Child Fear Survey schedule (DFSS_SF) was administered to 81 patients (first time attendees at the clinic) between the ages of eight and 13 years. The psychometric scale was used after determining its reliability and validity to ensure that it was culturally appropriate. Age, gender, sociodemographic status were also recorded. The dental anxiety score distribution, its threshold level as well as the relationship between the sociodemographic variables and patients' anxiety were determined. RESULTS: There was no statistically significant association between age (P = 0.856), gender (P = 0.124), socio-economic status of the children (P= 0.12) and level of dental anxiety. However, a statistically significant relationship did exist between the type of school the child attended and the anxiety level (P = 0.009). Also, 18 (14.8%) of the children were found to have high anxiety levels. CONCLUSIONS: Further studies involving larger non-clinic samples are needed to examine the issues raised by the findings of this study. PMID- 12542621 TI - Fluoride intake in children living in a high-fluoride area in Ethiopia - intake through beverages. AB - INTRODUCTION: The present study was conducted in Wonji Shoa, a sugar estate in the Ethiopian Rift Valley. Drinking water in the area is provided either by the Awash River or by high-fluoride ground water wells. Defluoridation plants have been installed, but are not in regular use, and fluorosis, dental as well as skeletal, is endemic. The aim of this study was to assess daily fluoride intake from drinking water and beverages in children from neighbouring villages with varying fluoride concentration in the drinking water. SUBJECTS AND METHODS: Thirty families were selected from two of the plantation villages (A and K). The criterion for being included in the project was the presence in the household of at least one child, fully weaned and below the age of 5 years. For sampling of beverages, the duplicate portion technique was used. The fluoride concentration in the beverage samples was determined using standard methods, using a fluoride ion-selective electrode. RESULTS: Ten of the selected households in Village A fetched water from the Awash River (1.8 mg F-/L) while five relied upon water from a local well (2.1 mg F-/L). All 15 households in Village K used water from a local well with fluoride concentration of 14.4 mg/L. The mean daily fluoride intake from drinking water and beverages during the four days, varied from 1.2 to 1.5 mg and 5.9 to 8.8 mg in Village A and K, respectively. Low variety in types of beverages consumed was reported both during the study period and through the questionnaire. Only local water was used for beverage preparation. Children who consumed milk had a reduced fluoride intake. Tea, which was part of the children's diet, was not found to be a main source of fluoride. CONCLUSION: A2n effective defluoridation of the drinking water or a change of water source would seem to be the only options for avoidance of dental and possibly skeletal fluorosis. PMID- 12542622 TI - Hypoplasia of primary and permanent teeth following osteitis and the implications of delayed diagnosis of a neonatal maxillary primary molar. AB - A 2-year-and-6-month-old Japanese girl with delayed eruption of a maxillary right primary canine and disturbance in the development of maxillary right primary molars was examined. The crown of the maxillary right first primary molar was severely decayed and hypoplastic. The periodontal tissues on the maxillary right posterior region were swollen and slight erythrogenic change was observed. Radiographic examination revealed a disturbance in the development of the permanent successor and the primary teeth in the maxillary right quadrant. According to the history, the maxillary right first primary molar had erupted on day seven after birth. It is suggested that the disturbance in development of the permanent successor and the primary teeth was secondary to osteitis caused by infection of the neonatal tooth. Appropriate dental treatment had not been administered for more than two years because the neonatal molar had not been diagnosed. PMID- 12542623 TI - Severe gingival hyperplasia in a child with I-cell disease. AB - I-cell disease (mucolipidosis II) is a rare metabolic disorder resulting from the deficiency of a specific lysosomal enzyme, N-acetylglucosamine-1 phosphotransferease. The disease presents as a mental and motor developmental delay with oral manifestations that include severe gingival hyperplasia usually seen before one year of age. The life expectancy of children with this condition is poor, with death usually occurring around the fifth year. A case report of a 3 year-old Pakistani male, with I-cell disease, is presented. The chief dental concerns of the parents were his swollen gums and delayed tooth eruption. Supportive treatment only was initiated. Differential diagnosis for severe gingival overgrowth in young patients should take account of this rare metabolic disorder in addition to hereditary and idiopathic fibromatosis and drug associated gingival overgrowth. PMID- 12542624 TI - Inverted eruption of a supplemental lower premolar: report of an unusual case. AB - Eruption of an inverted tooth is a rare condition. This case report describes the inverted eruption of a supplemental lower premolar in a 10-year-old Caucasian female. The tooth erupted in a downward direction and reached the lower cortical border of the mandible 2 years later. During an 8-year follow-up period, the tooth did not interfere with eruption of permanent teeth and the patient remained symptom-free. Considering clinical examination, medical and dental history as well as scientific evidence, the inverted eruption was most likely caused by a developmental abnormality with an inverted location of the tooth bud. This reported eruption of an inverted tooth is extremely rare and requires a radiographic follow-up to evaluate the relative position of this tooth in the jaw bone and its potential interference with neighbouring teeth or anatomic structures. PMID- 12542625 TI - Lesch-Nyhan syndrome: a preventive approach to self-mutilation. AB - Lesch-Nyhan syndrome (LNS), a rare inborn error of metabolism, is characterized by self-injurious behaviour, which results in partial or total destruction of oral and perioral tissues and/or fingers and hands. Prevention of self-mutilation raises significant difficulties. Treatment modalities include drug therapy, extraction of teeth, oral appliances and orthognathic surgery, all with variable success. A case of a 10-year-old boy with aggressive behaviour and severe lower lip injuries is presented. A palatal plate fabricated to raise the anterior bite proved to have satisfactory results. Although two adjustments were required during the recall visits, the effectiveness of the appliance in assisting healing of soft tissue damage was immediate and still apparent during the 3 years and 8 month follow-up period. PMID- 12542626 TI - A case of bilateral supplemental maxillary central incisors. AB - A case of bilateral supplemental maxillary central incisors is presented. Treatment comprised of extraction of one supplemental and two lateral incisors, preservation of one supplemental incisor, finishing with a 'Cyclops' arrangement of the teeth. The handling of supernumerary teeth in general is discussed. PMID- 12542627 TI - Lymphangioma of the tongue. PMID- 12542630 TI - Isolation and characterization of a termite transferrin gene up-regulated on infection. AB - PCR-based subtractive hybridization was used to isolate genes preferentially expressed in a termite (Mastotermes darwiniensis) following exposure to an entomopathogenic fungus. The subtraction procedure yielded a cDNA clone encoding a putative transferrin that, when sequenced to its ends, is the largest (728 amino acids) for any insect transferrin characterized to date. Cysteines and residues comprising putative iron-binding sites are conserved in both N- and C terminal lobes, suggesting structural and functional similarity to diferric vertebrate transferrins. A quantitative PCR assay confirmed a significant increase in transferrin expression following infection, suggesting its up regulation is part of the innate immune response. However, codon-based tests for selection among known insect transferrins revealed only a small proportion of codon-sites positively selected. Thus, unlike certain vertebrate transferrin lineages, no widespread evidence for pathogen-mediated positive selection was detected at this locus. PMID- 12542631 TI - Toxoneuron nigriceps polydnavirus encodes a putative aspartyl protease highly expressed in parasitized host larvae. AB - Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae) is an endophagous parasitoid of larval stages of the tobacco budworm, Heliothis virescens (F.) (Lepidoptera: Noctuidae). This parasitoid is associated with a polydnavirus (TnBV), injected at oviposition along with the egg, and involved in the disruption of host immune reaction and endocrine balance. This paper reports the molecular characterization of TnBV2, one of the most abundant genes in the TnBV genome. TnBV2 expression produces a mature 0.6 kb transcript in fat body, prothoracic glands and haemocytes, as early as 6 h after parasitoid oviposition. Only in haemocytes a specific longer transcript of 2.5 kb is found 24 h after parasitization. The putative translation product of TnBV2 contains a retroviral type aspartyl protease domain. The possible origin and functional role of this TnBV gene are discussed. PMID- 12542632 TI - Cloning and characterization of a 70 kDa heat shock cognate gene (HSC70) from two species of Chironomus. AB - In the present study we carried out the isolation and characterization of an HSC70 gene from two midges, Chironomus tentans and C. yoshimatsui. The HSC70 cDNAs are approximately 2424 (C. tentans) and 2464 bp (C. yoshimatsui) long, and contain 1950 and 1956 bp open reading frames, respectively. Analysis of genomic DNA revealed the presence of two introns in these genes. The 5' untranslated regions of the HSC70 genes are adenosine-rich, a feature found in inducible HSP70 genes. The nucleotide and amino acid sequences exhibit high identity with cytosolic HSC70s from other Dipterans. Northern hybridization indicated that HSC70 is expressed at all developmental stages, from embryo to adult, and Southern hybridization confirmed the presence of multiple HSP70 genes in Chironomus. PMID- 12542633 TI - Functional analysis of a G protein-coupled receptor from the southern cattle tick Boophilus microplus (Acari: Ixodidae) identifies it as the first arthropod myokinin receptor. AB - The myokinins are invertebrate neuropeptides with myotropic and diuretic activity. The lymnokinin receptor from the snail Lymnaea stagnalis (Mollusca) has been the only previously identified myokinin receptor. We had cloned a G protein coupled receptor (AF228521) from the tick Boophilus microplus (Arthropoda: Acari), 40% identical to the lymnokinin receptor, that we have now expressed in CHO-K1 cells. Myokinins at nanomolar concentrations induced intracellular calcium release, as measured by fluorescent cytometry and the receptor coupled to a pertussis toxin-insensitive G protein. Absence of extracellular calcium did not inhibit the fluorescence response, indicating that intracellular stores were sufficient for the initial response. Control cells only transfected with vector did not respond. We conclude that the tick receptor is the first myokinin receptor to be cloned from an arthropod. PMID- 12542634 TI - Cytochrome P450 CYP6X1 cDNAs and mRNA expression levels in three strains of the tarnished plant bug Lygus lineolaris (Heteroptera: Miridae) having different susceptibilities to pyrethroid insecticide. AB - Three cDNAs, cloned from both pyrethroid-susceptible and -resistant strains of Lygus lineolaris, contained a 1548 nucleotide open reading frame encoding a 516 amino acid residue protein. Predicted cytochrome P450s from cDNAs were classified as the first three new members of subfamily CYP6X, CYP6X1v1 for a susceptible strain and CYP6X1v2 and CYP6X1v3 for two resistant strains. Putative cytochrome P450 CYP6X1s from L. lineolaris were highly similar (up to 42% amino acid sequence identity) to several insect CYP6s that are responsible for reduced sensitivity to pyrethroid insecticides. A total of twenty-six nucleotide substitutions were revealed between cDNAs of susceptible and resistant strains. Two nucleotide substitutions resulted in amino acid changes, Asp373 to Ala373 and Ser487 to Ala487, between susceptible and resistant strains. The resistant laboratory strain contained 2.1-fold higher cytochrome P450 mRNA per microgram total RNA than the susceptible laboratory strain. Topical treatment with 10 ng permethrin elevated cytochrome P450 mRNA levels by approximately 2-fold. The results of this study indicated that cytochrome P450 gene mutation, coupled with up-regulation, was present only in the pyrethroid resistant strains, and was possibly related to resistance development in the tarnished plant bug. PMID- 12542635 TI - Isolation and expression of a sterol carrier protein-2 gene from the yellow fever mosquito, Aedes aegypti. AB - Trafficking of cholesterol in insects is a very important process due to the fact that insects depend on dietary cholesterol to fulfil their physiological needs. We identified a putative mosquito sterol carrier protein-2 (SCP-2) cDNA from fourth instar subtracted cDNA library. The AeSCP-2 protein has high degree homology in the sterol transfer domain to both rat and human SCP-2. Transcripts of AeSCP-2 in fourth instars were detected strongly in the midgut, and weakly in the head and hindgut. In the early pupae, AeSCP-2 transcription was observed in the thorax, head and body wall of abdomen, but not in the gut. The interaction of mosquito sterol carrier protein-2 (AeSCP-2) with cholesterol was examined. The Kd of purified recombinant AeSCP-2 to cholesterol was 5.6 +/- 0.6 x 10-9 m using radiolabelled cholesterol-binding assay. The results suggest that AeSCP-2 has high affinity to cholesterol and may function as a carrier protein in mosquitoes. PMID- 12542636 TI - Characterization of JNK-like protein derived from a mosquito cell line, C6/36. AB - When Western blot analysis of heat-killed bacteria- and lipopolysaccharide (LPS) treated Aedes albopictus mosquito cell line C6/36 was performed using antiphospholyrated c-Jun amino-terminal kinase (JNK) antibodies, approximately 46 kDa protein was clearly detected with a peak around 30 min. After the C6/36 cells were incubated at 45 degrees C in order to induce apoptosis, the 46 kDa protein continued to be detected for at least 3 h. The internalization of fluorescein labelled bacteria was inhibited by a JNK-specific inhibitor SP600125, suggesting that phagocytosis involves the JNK signalling pathway in mosquito cells. Based on these results, we found one candidate for the nucleotide sequence of JNK (Ae-JNK) from the C6/36 cells. This study is the first report regarding the mitogen activated protein kinase (MAPK) of mosquito. PMID- 12542637 TI - Characterization of three Toll-like genes from mosquito Aedes aegypti. AB - Three Toll-related genes (AeToll1A, AeToll1B and AeToll5) were cloned and characterized from the yellow fever vector mosquito, Aedes aegypti. All three genes exhibited high levels of amino acid sequence similarity with Drosophila melanogaster (Dm)Toll1 and DmTehao (Toll5). AeToll1A and AeToll1B are 1124 and 1076 amino acid residues long, respectively. Both contain a carboxyl extension downstream of the Toll/interleukin-1 receptor (TIR) domain. AeToll5 is 1007 residues long and, like DmTehao, lacks the carboxyl terminal extension. Expression of these three genes was examined throughout development and after immune challenge. Both AeToll1A and AeToll5, like their Drosophila counterparts, activate transcription of drosomycin promoter in both Aedes and Drosophila cell lines. Deletion of the carboxyl extension of AeToll1A did not result in a further elevated level of the antifungal response. The intracellular signalling process appears to be species specific based on two observations. (1) DmToll is completely inactive in an Aedes cell line, suggesting a higher specificity requirement for DmToll in the intracellular signalling process. (2) Only one of three amino acid residues essential for DmToll function is required for AeToll1A function. PMID- 12542638 TI - Translation of both 5'TOP and non-TOP host mRNAs continues into the late phase of Baculovirus infection. AB - Complete cDNA sequences were obtained for ribosomal protein (rp) L15 and eukaryotic initiation factor eIF2alpha from the lepidopteran insect Spodoptera frugiperda, and for elongation factor eEF2 from S. exigua. The presence of a 5' terminal oligopyrimidine (TOP) tract classified the lepidopteran rpL15 transcript as a TOP mRNA. For eEF2, two types of transcripts were observed, one of which had a 5'TOP tract. The transcript levels for rpL15, eEF2 and eIF2alpha decreased following baculovirus infection. Polysome analysis showed that the corresponding mRNAs remained to be translated until at least 16 h post-infection for both TOP and non-TOP mRNAs. Baculovirus-induced host shut-off therefore appears to be regulated at the level of RNA abundance rather than at the translational level. PMID- 12542639 TI - Application of 16S rDNA-DGGE to examine the microbial ecology associated with a social wasp Vespula germanica. AB - Invertebrates host numerous bacteria, with interactions ranging from pathogenesis to symbiosis. While certain symbiotic relationships have been well studied, little is known about the dynamics of these bacterial communities. Denaturing gradient gel electrophoresis (DGGE) was used to examine the bacterial microflora associated with the eusocial wasp Vespula germanica. DGGE profiles of larval guts revealed a variable microflora, suggesting that V. germanica is not dependent on a particular suite of mutualists. The variation in profiles was not related to season, nest size or macrohabitat. Sequences corresponding to Lactococcus, Lactobacillus, a novel Leuconostoc and two Rickettsiella grylli strains were obtained. DGGE proved to be a useful technique for characterizing the wasp microflora. Given the importance of microbial communities to invertebrates, there is much to be gained from the application of such techniques. PMID- 12542640 TI - Hidden from the host: Spiroplasma bacteria infecting Drosophila do not cause an immune response, but are suppressed by ectopic immune activation. AB - Insects and other arthropods have an effective innate immune system that can clear infections with bacteria and other microorganisms. Despite this ability, one group of bacteria, the spiroplasmas, survive unharmed within the haemolymph of a wide range of arthropod hosts. We investigated the interaction between one member of this clade, a relative of Spiroplasma poulsonii, and the immune system of its Drosophila host. Expression of antimicrobial genes in spiroplasma-infected flies did not differ from wild-type controls either in the naturally infected state, nor after septic shock. We therefore concluded that spiroplasma infection did not induce an immune response in its host, but that this absence of response was unlikely to be because the bacterium inhibited response. Further experiments revealed immune reactions induced ectopically did reduce parasite titre. We therefore conclude that this bacterium has a novel form of interaction with its host, being hidden from the host immune system, but potentially suppressible by it. PMID- 12542655 TI - The mouse VPAC2 receptor confers suprachiasmatic nuclei cellular rhythmicity and responsiveness to vasoactive intestinal polypeptide in vitro. AB - Expression of coherent and rhythmic circadian (approximately 24 h) variation of behaviour, metabolism and other physiological processes in mammals is governed by a dominant biological clock located in the hypothalamic suprachiasmatic nuclei (SCN). Photic entrainment of the SCN circadian clock is mediated, in part, by vasoactive intestinal polypeptide (VIP) acting through the VPAC2 receptor. Here we used mice lacking the VPAC2 receptor (Vipr2-/-) to examine the contribution of this receptor to the electrophysiological actions of VIP on SCN neurons, and to the generation of SCN electrical firing rate rhythms SCN in vitro. Compared with wild-type controls, fewer SCN cells from Vipr2-/- mice responded to VIP and the VPAC2 receptor-selective agonist Ro 25-1553. By contrast, similar proportions of Vipr2-/- and wild-type SCN cells responded to gastrin-releasing peptide, arginine vasopressin or N-methyl-D-aspartate. Moreover, VIP-evoked responses from control SCN neurons were attenuated by the selective VPAC2 receptor antagonist PG 99-465. In firing rate rhythm experiments, the midday peak in activity observed in control SCN cells was lost in Vipr2-/- mice. The loss of electrical activity rhythm in Vipr2-/- mice was mimicked in control SCN slices by chronic treatment with PG 99-465. These results demonstrate that the VPAC2 receptor is necessary for the major part of the electrophysiological actions of VIP on SCN cells in vitro, and is of fundamental importance for the rhythmic and coherent expression of circadian rhythms governed by the SCN clock. These findings suggest a novel role of VPAC2 receptor signalling, and of cell-to-cell communication in general, in the maintenance of core clock function in mammals, impacting on the cellular physiology of SCN neurons. PMID- 12542656 TI - Neurogenesis in explants from the walls of the lateral ventricle of adult bovine brain: role of endogenous IGF-1 as a survival factor. AB - Previous studies have shown the existence of proliferating cells in explants from bovine (Bos Taurus) lateral ventricle walls that were maintained for several days in vitro in the absence of serum and growth factors. In this study we have characterized the nature of new cells and have assessed whether the insulin-like growth factor-1 (IGF-1) receptor regulates their survival and/or proliferation. The explants were composed of the ependymal layer and attached subependymal cells. Ependymal cells in culture were labelled with glial markers (S-100, vimentin, GFAP, BLBP, 3A7 and 3CB2) and did not incorporate bromodeoxiuridine when this molecule was added to the culture media. Most subependymal cells were immunoreactive for beta III-tubulin, a neuronal marker, and did incorporate bromodeoxiuridine. Subependymal neurons displayed immunoreactivity for IGF-1 and its receptor and expressed IGF-1 mRNA, indicating that IGF-1 is produced in the explants and may act on new neurons. Addition to the culture media of an IGF-1 receptor antagonist, the peptide JB1, did not affect the incorporation of bromodeoxiuridine to proliferating subependymal cells. However, JB1 significantly increased the number of TUNEL positive cells in the subependymal zone, suggesting that IGF-1 receptor is involved in the survival of subependymal neurons. In conclusion, these findings indicate that neurogenesis is maintained in explants from the lateral cerebral ventricle of adult bovine brains and that IGF-1 is locally produced in the explants and may regulate the survival of the proliferating neurons. PMID- 12542658 TI - Suprachiasmatic control of melatonin synthesis in rats: inhibitory and stimulatory mechanisms. AB - The suprachiasmatic nucleus (SCN) controls the circadian rhythm of melatonin synthesis in the mammalian pineal gland by a multisynaptic pathway including, successively, preautonomic neurons of the paraventricular nucleus (PVN), sympathetic preganglionic neurons in the spinal cord and noradrenergic neurons of the superior cervical ganglion (SCG). In order to clarify the role of each of these structures in the generation of the melatonin synthesis rhythm, we first investigated the day- and night-time capacity of the rat pineal gland to produce melatonin after bilateral SCN lesions, PVN lesions or SCG removal, by measurements of arylalkylamine N-acetyltransferase (AA-NAT) gene expression and pineal melatonin content. In addition, we followed the endogenous 48 h-pattern of melatonin secretion in SCN-lesioned vs. intact rats, by microdialysis in the pineal gland. Corticosterone content was measured in the same dialysates to assess the SCN lesions effectiveness. All treatments completely eliminated the day/night difference in melatonin synthesis. In PVN-lesioned and ganglionectomised rats, AA-NAT levels and pineal melatonin content were low (i.e. 12% of night-time control levels) for both day- and night-time periods. In SCN lesioned rats, AA-NAT levels were intermediate (i.e. 30% of night-time control levels) and the 48-h secretion of melatonin presented constant levels not exceeding 20% of night-time control levels. The present results show that ablation of the SCN not only removes an inhibitory input but also a stimulatory input to the melatonin rhythm generating system. Combination of inhibitory and stimulatory SCN outputs could be of a great interest for the mechanism of adaptation to day-length (i.e. adaptation to seasons). PMID- 12542657 TI - Expression of the circadian clock gene Period 1 in neuroendocrine cells: an investigation using mice with a Per1::GFP transgene. AB - The circadian clock located in the suprachiasmatic nuclei (SCN) of the hypothalamus regulates daily temporal organization in behaviour and neuroendocrine function. The molecular basis for circadian rhythm generation is an interacting transcriptional/translational feedback loop comprised of several 'clock genes' and their respective protein products. Clock genes are expressed not only in the SCN but also in numerous other locations throughout the brain, including regions rich in neuroendocrine cells. In order to investigate whether neuroendocrine cells function as autonomous oscillators, we used female transgenic mice in which an unstable, degradable jellyfish green fluorescent protein (GFP) gene is driven by a mouse Period 1 (Per1) gene promoter. Mice were injected (s.c.) with fluorogold (FG) in order to label neuroendocrine cells and brain sections were double-labelled for either FG and Per1 mRNA (labelled by GFP immunostaining) or FG and PER1 protein using fluorescence immunocytochemistry. Mice were killed during either the day or night. Neuroendocrine cells contained Per1 mRNA and PER1 protein in several brain regions with the greatest proportion of double-labelled cells occurring in the arcuate nucleus (Arc). The number of neuroendocrine cells labelled was not affected by the stage of the estrous cycle. Fewer FG-labelled cells expressed Per1 message and protein during the night compared to the day. In the Arc, staining for tyrosine hydroxylase revealed that neuroendocrine cells expressing Per1 message and protein were dopaminergic. Together, these findings suggest that neuroendocrine cells contain the molecular machinery necessary to oscillate independently. It remains to be determined whether these cells actually function as autonomous oscillators or whether these rhythms are driven by signals from the SCN. These findings also indicate that the endocrine system represents an opportunity to study the interactions between central (SCN and neuroendocrine cells) and peripheral circadian (endocrine gland) oscillators. PMID- 12542659 TI - Neurons of the trigeminal main sensory nucleus participate in the generation of rhythmic motor patterns. AB - The trigeminal principal sensory nucleus (NVsnpr) contains both trigemino thalamic neurons and interneurons projecting to the reticular formation and brainstem motor nuclei. Here we describe the inputs and patterns of firing of NVsnpr neurons during fictive mastication in anaesthetized and paralysed rabbits to determine the role that NVsnpr may play in patterning mastication. Of the 272 neurons recorded in NVsnpr, 107 changed their firing patterns during repetitive stimulation of the left or right sensorimotor cortex to induce fictive mastication. Thirty increased their firing tonically. Seventy-seven became rhythmically active, but only 31 fired in phase with mastication. The others discharged in bursts at more than twice the frequency of trigeminal motoneurons. Most rhythmic masticatory neurons were concentrated in the dorsal part, and those which fired during the jaw closing phase of the cycle were confined to the anterior pole of the nucleus. Most of these cells had inputs from muscle spindle afferents, whereas most of those firing during jaw opening had inputs from periodontal receptors. Non-masticatory rhythmical neurons had receptive fields on the lips and face. The majority of rhythmical masticatory units were modulated during fictive mastication evoked by both the left and right cortices and only four changed their phase of firing when switching from one cortex to the other. When coupled with the finding that NVsnpr neurons exhibit spontaneous bursting in vitro[Sandler et al. (1998) Neuroscience, 83, 891], the results described here suggest that neurons of dorsal NVsnpr may form the core of the central pattern generator for mastication. PMID- 12542660 TI - Localization of a FMRFamide-related peptide in efferent neurons and analysis of neuromuscular effects of DRNFLRFamide (DF2) in the crustacean Idotea emarginata. AB - In the ventral nerve cord of the isopod Idotea emarginata, FMRFamide immunoreactive efferent neurons are confined to pereion ganglion 5 where a single pair of these neurons was identified. Each neuron projects an axon into the ipsilateral ventral and dorsal lateral nerves, which run through the entire animal. The immunoreactive axons form numerous varicosities on the ventral flexor and dorsal extensor muscle fibres, and in the pericardial organs. To analyse the neuromuscular effects of a FMRFamide, we used the DRNFLRFamide (DF2). DF2 acted both pre- and postsynaptically. On the presynaptic side, DF2 increased transmitter release from neuromuscular endings. Postsynaptically, DF2 depolarized muscle fibres by approximately 10 mV. This effect was not observed in leg muscles of a crab. The depolarization required Ca2+, was blocked by substituting Ca2+ with Co2+, but not affected by nifedipine or amiloride. In Idotea, DF2 also potentiated evoked extensor muscle contractions. The amplitude of high K+ contractures was increased in a dose dependent manner with an EC50 value of 40 nm. In current-clamped fibres, DF2 strongly potentiated contractions evoked by current pulses exceeding excitation-contraction threshold. In voltage-clamped fibres, the inward current through l-type Ca2+ channels was increased by the peptide. The observed physiological effects together with the localization of FMRFamide-immunoreactive efferent neurons suggest a role for this type of peptidergic modulation for the neuromuscular performance in Idotea. The pre- and postsynaptic effects of DF2 act synergistically and, in vivo, all should increase the efficacy of motor input to muscles resulting in potentiation of contractions. PMID- 12542661 TI - Muscarinic and GABAA receptors modulate acetylcholine release in feline basal forebrain. AB - Acetylcholine (ACh) release within the basal forebrain changes significantly as a function of sleep and wakefulness, hence identifying the neurochemical modulators of basal forebrain ACh release will contribute to a mechanistic understanding of sleep cycle regulation. This study tested the hypothesis that muscarinic and gamma aminobutyric acid(A) (GABAA) receptors modulate basal forebrain ACh release. Cats were anaesthetized with halothane to hold arousal state constant and a microdialysis probe was aimed stereotaxically for the substantia innominata region of the basal forebrain. Four concentrations of the muscarinic antagonist scopolamine (0.1, 0.3, 1.0, and 10 nm) and five concentrations of the GABAA antagonist bicuculline (3, 10, 30, 100, and 300 micro m) were delivered by reverse dialysis from the same probes used to collect ACh. These results are based on 27 experiments in nine animals. Scopolamine and bicuculline each caused a concentration dependent enhancement of ACh release. Scopolamine increased ACh by 118% above control levels whereas bicuculline was more effective, causing a 287% increase in ACh release. Scopolamine was more potent (EC50 = 0.16 nm) than bicuculline (EC50 > or = 90 micro m) for increasing ACh release. The results support the hypothesis that substantia innominata ACh release is modulated by muscarinic autoreceptors and inhibited by GABAA receptors. These findings are consistent with the interpretation that inhibition of basal forebrain cholinergic neurotransmission by GABA contributes to the generation of sleep. PMID- 12542662 TI - Long-term striatal overexpression of GDNF selectively downregulates tyrosine hydroxylase in the intact nigrostriatal dopamine system. AB - Sustained neurotrophic factor treatment in neurodegenerative disorders such as Parkinson's disease is likely to affect both degenerating and intact neurons. To investigate the effect of long-term glial cell line-derived neurotrophic factor (GDNF) overexpression on intact nigrostriatal dopamine neurons, we injected a recombinant lentiviral vector encoding GDNF, or green fluorescent protein, in the right striatum of young adult rats. Thirteen months after viral injection GDNF levels were 4.5 ng/mg tissue in the striatum and 0.9 ng/mg in the substantia nigra as measured by ELISA, representing a 25-100-fold increase above control vector- or nontransduced tissue. GDNF overexpression significantly reduced tyrosine hydroxylase mRNA levels (by 39-72%) in the substantia nigra and ventral tegmental area neurons, and the optical density of tyrosine hydroxylase immunoreactive innervation in the striatum was reduced by 25-52% with the most prominent reductions appearing caudally. No significant reduction was seen in striatal vesicular monoamine transporter 2-immunoreactivity or [3H]mazindole binding autoradiography to dopamine uptake sites, two other presynaptic markers in dopamine axon terminals. The striatal D1 and D2 receptor binding as determined by [3H]SCH23390 and [3H]spiperone binding, respectively, was unaltered relative to the intact side in both treatment groups. Preproenkephalin mRNA levels in postsynaptic striatal neurons, which increase upon removal of striatal dopamine, were also unaffected by the GDNF treatment. Taken together our findings indicate that sustained GDNF administration to intact nigrostriatal dopamine neurons selectively reduces tyrosine hydroxylase expression, without altering striatal dopamine transmission to the extent that compensatory changes in several other components related to dopamine storage and signalling occur. PMID- 12542663 TI - cDNA profiling of epileptogenesis in the rat brain. AB - Symptomatic temporal lobe epilepsy typically develops in three phases: brain insult --> latency period (epileptogenesis) --> recurrent seizures (epilepsy). We hypothesized that remodeling of neuronal circuits underlying epilepsy is associated with altered gene expression during epileptogenesis. Epileptogenesis was induced by electrically triggered status epilepticus (SE) in rats. Animals were continuously monitored with video-EEG, and the hippocampus and temporal lobe were collected either during epileptogenesis (1, 4 and 14 days) or after the first spontaneous seizures (14 days) for cDNA array analysis. Altogether, 282 genes had altered expression, from which 87 were in the hippocampus and 208 in the temporal lobe (overlap in 13). Assessment of hippocampal gene expression during epileptogenesis indicated that 37 genes were altered in the 1-day group, 12 in the 4-day group and 14 in the 14-day epileptogenesis group. There were 42 genes with altered expression in the 14-day epilepsy group. In the temporal lobe, the number of genes with altered expression was 29 in the 1-day group, 155 in the 4-day group, 32 in the 14-day epileptogenesis group and 62 in the 14-day epilepsy group. Products of the altered genes are involved in neuronal plasticity, gliosis, organization of the cytoskeleton or extracellular matrix, cell adhesion, signal transduction, regulation of cell cycle, and metabolism. As most of these genes have not previously been implicated in epileptogenesis or epilepsy, these data open new avenues for understanding the molecular basis of epileptogenesis and provide new targets for rational development of anti-epileptogenic treatments for patients with an elevated risk of epileptogenesis after brain injury. PMID- 12542664 TI - Electrophysiological evidence for convergence of inputs from the medial prefrontal cortex and lateral habenula on single neurons in the dorsal raphe nucleus. AB - Neuronal projections to the dorsal raphe nucleus (DRN) from the medial prefrontal cortex (mPFC) and lateral habenula nucleus (LHb) provide the two key routes by which information processed by mood regulatory, cortico-limbic-striatal circuits input into the 5-HT system. These two projections may converge as it appears that both activate local GABAergic neurons to inhibit 5-HT neurons in the DRN. Here we have tested this hypothesis by measuring the effect of stimulation of the mPFC and LHb on the activity of 5-HT and non-5-HT, putative gamma-amino butyric acid (GABA) neurons in the DRN using extracellular recordings in anaesthetized rats. A total of 119 5-HT neurons (regular, slow firing, broad spike width) and 21 non-5 HT, putative GABA neurons (fast-firing, narrow spike width) were tested. Electrical stimulation of the mPFC or LHb caused a poststimulus inhibition (30 ms latency) of 101/119 5-HT neurons, of which 61 (60%) were inhibited by both the mPFC and LHb. Electrical stimulation of the mPFC or LHb also caused a short latency (12-20 ms) poststimulus facilitation of 10/21 non-5-HT neurons, of which 5 (50%) were activated by both the mPFC and LHb. These data indicate that a significant number of 5-HT neurons and non-5-HT neurons in the DRN are influenced by both the mPFC and LHb. Moreover, the data are compatible with the hypothesis and that there is a convergence of mPFC and LHb inputs on local circuit GABAergic neurons in the DRN which in turn inhibit the activity of 5-HT neurons. PMID- 12542665 TI - Imaging of calcineurin activated by long-term depression-inducing synaptic inputs in living neurons of rat visual cortex. AB - Long-term depression (LTD) of synaptic transmission is induced by low-frequency stimulation (LFS) of afferents lasting for a long time, typically for 10-15 min, in neocortical and hippocampal slices. It is suggested that calcineurin, Ca2+/calmodulin-dependent protein phosphatase, plays a role in the induction of LTD, based on the results that pharmacological or genetic manipulation of calcineurin activity interfered in its induction. However, questions as to why it takes so long to induce LTD and in which compartment of neurons calcineurin is activated remain unanswered. With a fluorescent indicator for calcineurin activity, we visualized the spatiotemporal pattern of its activation in living neurons in layer II/III of visual cortical slices of rats during the LFS of layer IV that induced LTD of synaptic responses. During LFS, the fluorescence intensity gradually increased with a latency of a few minutes in dendrites and soma of neurons, and remained increased during the whole observation period (10-25 min) after LFS. The onset latency of the increase in the soma was slower than that in the distal dendritic region. The LFS-induced rise in fluorescence was not observed in neurons which were loaded with inhibitors of calcineurin, indicating that the intensity of fluorescence reflects calcineurin activity. Control stimulation at 0.05 Hz and theta-burst stimulation did not significantly change the intensity of fluorescence. Only LFS-type inputs effectively activate calcineurin in postsynaptic neurons in an augmenting manner, and such a time consuming activation of calcineurin may be a reason why long-lasting LFS is necessary for the induction of LTD. PMID- 12542666 TI - Substance induced plasticity in noradrenergic innervation of the paraventricular hypothalamic nucleus. AB - Single administration of the cytokine interleukin-1 alpha (IL-1), or the psychostimulant amphetamine, enhanced adrenocorticotropin hormone and corticosterone responses to a stress challenge weeks later. This long-lasting hypothalamic-pituitary-adrenal (HPA)-sensitization is paralleled by an increase in electrically evoked release of noradrenaline in the paraventricular hypothalamic nucleus (PVN). We hypothesized that these functional changes may be associated with morphological plasticity of noradrenergic projections to the PVN, a parameter that shows high reproducibility. Specific alterations in relative (nor)adrenergic innervation density were studied by using dopamine-alpha hydroxylase (DBH) as a marker. An image analysis system was used to detect changes in the relative DBH innervation density of the PVN. Groups of adult male rats were given IL-1 (10 microg/kg i.p.), amphetamine (5 mg/kg i.p.), or saline. Three weeks later, IL-1 and amphetamine primed rats showed enhanced adrenocorticotropin hormone and corticosterone responses to an amphetamine challenge. In another set of experiments, the relative DBH innervation density was measured in different PVN subnuclei at four rostro-caudal levels. Single administration of either IL-1 or amphetamine causes three weeks later a selective decrease in relative DBH innervation density in those subnuclei of the PVN that contain high numbers of corticotrophin-releasing hormone (CRH) producing neurons: the dorsal parvocellular and medial parvocellular PVN. We conclude that (1) long lasting sensitization induced by single exposure to IL-1 and amphetamine induces specific pattern of neuroplastic changes in (nor)adrenergic innervation in the PVN and (2) reduction of relative DBH innervation density in CRH-rich areas is associated with paradoxical increase of electrically evoked release of (nor)adrenaline. PMID- 12542667 TI - Ciproxifan, a histamine H3-receptor antagonist/inverse agonist, modulates the effects of methamphetamine on neuropeptide mRNA expression in rat striatum. AB - We have explored the effect of histamine H3-receptor ligands on the regulation of neuropeptide mRNA expression in the striatum by using in situ hybridization performed with proenkephalin, prodynorphin, substance P and proneurotensin riboprobes. Acute administration of ciproxifan, an H3-receptor antagonist/inverse agonist, or (R)-alpha-methylhistamine, an H3-receptor agonist, did not modify the striatal expression of the neuropeptides by itself. However, ciproxifan strongly and differentially modulated the effect of a single administration of 3 mg/kg methamphetamine on neuropeptide mRNA expression. This modulation was suppressed by the administration of (R)-alpha-methylhistamine and occurred in both the caudate-putamen and nucleus accumbens. Ciproxifan strongly potentiated the decrease of proenkephalin mRNA expression induced by methamphetamine. In contrast, it suppressed the increase in prodynorphin and substance P mRNA expression induced by methamphetamine. Methamphetamine alone or with ciproxifan did not modify proneurotensin mRNA expression. These neurochemical findings indicate that ciproxifan differentially regulates the effect of methamphetamine on the neuropeptides contained in striatonigral and striatopallidal neurons. They suggest that endogenous histamine and dopamine cooperate to modulate the activity of striatal projection neurons and strengthen the interest of H3-receptors as new targets for the treatment of psychotic disorders and drug abuse. PMID- 12542669 TI - Cellular mechanisms of Delta 9-tetrahydrocannabinol behavioural sensitization. AB - We investigated the cellular events linked to the induction of cannabinoid behavioural sensitization. In sensitized rats, autoradiographic binding studies with [3H]CP-55,940 showed a significant increase in cannabinoid receptor binding, specifically in the cerebellum, with no changes in the other brain areas where basal CB1-receptor expression is observed. In vitro autoradiography of CP-55,940 stimulated [35S]GTP gamma S binding provided a picture of cannabinoid receptor mediated G protein activation. Basal [35S]GTP gamma S binding was not affected, whereas sensitized rats showed a significant increase of net [35S]GTP gamma S binding in the caudate putamen and cerebellum. Autoradiographic studies suggested that only these two areas had altered receptor functionality. We therefore focused our intracellular investigations only there, first surveying the responsiveness of the cAMP system to cannabinoids. CP-55,940 was unable to inhibit forskolin-induced cAMP accumulation in the cerebellum of sensitized animals, but no difference was observed between groups in the caudate putamen. Finally, we surveyed the levels of CREB and AP-1 binding activity, in the same two areas and found no difference in sensitized rats. The intracellular picture in sensitized rats suggests that besides the cAMP cascade, other signalling pathways may participate in the development of cannabinoid sensitization. PMID- 12542668 TI - Increase of morphine withdrawal in mice lacking A2a receptors and no changes in CB1/A2a double knockout mice. AB - CB1 cannabinoid and A2a adenosine receptors are highly expressed in the central nervous system where they modulate numerous physiological processes including emotional behaviour and the responses of several drugs of abuse. To investigate the contribution of these receptors in emotional-like responses and opioid dependence we have generated CB1/A2a double deficient mice (CB1-/-/A2a-/-). The spontaneous locomotor activity was reduced in double knockout as compared to wild type animals. Emotional-like responses of CB1-/-/A2a-/- mice were investigated using the elevated plus-maze and the lit-dark box. Mutant mice exhibited an increased level of anxiety in both behavioural models. The specific involvement of CB1 and A2a receptors in morphine dependence was evaluated by using A2a knockout mice and CB1/A2a double mutant mice. The severity of naloxone precipitated morphine withdrawal syndrome was significantly increased in the absence of A2a adenosine receptors whereas no modifications were observed in the double knockout mice. These results suggest that both receptors participate in the control of emotional behaviour and seem to play an opposite role in the expression of opioid physical dependence. PMID- 12542670 TI - LTP but not seizure is associated with up-regulation of AKAP-150. AB - We have used differential display to profile and compare the mRNAs expressed in the hippocampus of freely moving animals after the induction of long-term potentiation (LTP) at the perforant path-dentate gyrus synapse with control rats receiving low-frequency stimulation. We have combined this with in situ hybridization and have identified A-kinase anchoring protein of 150 kDa (AKAP 150) as a gene selectively up-regulated during the maintenance phase of LTP. AKAP 150 mRNA has a biphasic modulation in the dentate gyrus following the induction of LTP. The expression of AKAP-150 was 29% lower than stimulated controls 1 h after the induction of LTP. Its expression was enhanced 3 (50%), 6 (239%) and 12 h (210%) after induction, returning to control levels by 24 h postinduction. The NMDA receptor antagonist CPP blocked the tetanus-induced modulation of AKAP-150 expression. Interestingly, strong generalized stimulation produced by electroconvulsive shock did not increase the expression of AKAP-150. This implies that the AKAP-150 harbours a novel property of selective responsiveness to the stimulation patterns that trigger NMDA-dependent LTP in vivo. Its selective up regulation during LTP and its identified functions as a scaffold for protein kinase A, protein kinase C, calmodulin, calcineurin and ionotropic glutamate receptors suggest that AKAP-150 encodes is an important effector protein in the expression of late LTP. PMID- 12542671 TI - Long-term potentiation in the hippocampal CA1 area and dentate gyrus plays different roles in spatial learning. AB - NMDA receptor-dependent long-term potentiation (LTP) at hippocampal synapses has been considered a crucial component of the cellular basis for learning and memory. This form of LTP occurs in excitatory synapses in both the CA1 area and the dentate gyrus in the hippocampus. However, differential roles of LTP in these areas have not yet been identified. To address this issue, we enhanced the degree of LTP by expressing Ca2+-permeable AMPA receptors at either hippocampal CA1 or dentate gyrus synapses using Sindbis viral vectors (SINs) encoding both green fluorescent proteins and unedited GluR2 (GluR2Q) subunits, and examined their effects on rat spatial learning. The viral vectors were locally injected into the 8-week-old-rat brain in vivo bilaterally. The postsynaptic expression of Ca2+ permeable AMPA receptors enhanced the degree of LTP, and induced NMDA receptor independent LTP in the presence of the NMDA receptor antagonist in SIN-infected regions in both CA1 and dentate gyrus in hippocampal slice preparations. However, the regional expression of Ca2+-permeable AMPA receptors caused opposite behavioural consequences on the Morris water maze task: rats with SIN-infected CA1 pyramidal cells showed shorter escape latency and better probe test performance, whereas those with SIN-infected dentate gyrus granule cells showed impaired performance. Thus, it was demonstrated that CA1 and dentate gyrus synapses play different functional roles in spatial learning despite their similar mechanism for LTP induction. PMID- 12542672 TI - Olfactory learning modifies the expression of odour-induced oscillatory responses in the gamma (60-90 Hz) and beta (15-40 Hz) bands in the rat olfactory bulb. AB - This study addressed the question of the possible functional relevance of two different oscillatory activities, beta and gamma (15-40 and 60-90 Hz, respectively) for perception and memory processes in olfactory areas of mammals. Local field potentials were recorded near relay olfactory bulb neurons while rats performed an olfactory discrimination task. Signals reflected the mass activity from this region and characteristics of oscillatory activities were used as an index of local synchrony. Beta and gamma oscillatory activities were quantified by time-frequency methods before during and after odour sampling. In rats early in their training, olfactory sampling was associated with a significant decrease in power in the gamma band in parallel with a weak but significant increase in the beta band (centred on 27 Hz). Several days later, in well-trained rats, the gamma oscillatory depression was significantly enhanced both in duration and amplitude. It appeared within the 500 ms time period preceding odour onset and was further reduced during the odour period. Concurrently the beta oscillatory response (now centred on 24 Hz) during odour sampling was amplified by a twofold factor. The beta band response was modulated according to the chemical nature of the stimuli and rat's behavioural response. This study showed for the first time that odour sampling in behaving animals is associated with a clear shift in the olfactory bulb neuronal activity from a gamma to a beta oscillatory regime. Moreover, the data stress the importance of studying the odour-induced beta activity and its relation to perception and memory. PMID- 12542673 TI - Activation of the phasic pontine-wave generator enhances improvement of learning performance: a mechanism for sleep-dependent plasticity. AB - The aim of this study was to test the hypothesis that supplementary activation of the phasic pontine wave (P-wave) generator during rapid eye movement (REM) sleep enhances consolidation and integration of memories, resulting in improved learning. To test this hypothesis, two groups of rats were trained on a two-way active avoidance learning task in the morning. Immediately after training, one group of rats received a carbachol microinjection into the P-wave generator and the other group was microinjected with control saline into the same target area. After training trials and microinjections, rats were allowed a 6-h period of undisturbed sleep in the polygraphic recording chamber. At the end of 6 h of undisturbed sleep-wake recordings, rats were retested in a session of avoidance learning trials. After learning trials, the total percentage of time spent in REM sleep was significantly increased in both saline (15.36%)- and carbachol (17.70%) microinjected rats. After learning trials, REM sleep P-wave density was significantly greater throughout the 6-h period of recordings in carbachol treated rats than in the saline treated rats. In the retrial session, the improvement in learning task performance was 22.75% higher in the carbachol microinjected rats than in the saline-microinjected rats. These findings show that the consolidation and integration of memories create a homeostatic demand for P-waves. In addition, these findings provide experimental evidence, for the first time, that activation of the P-wave generator may enhance consolidation and integration of memories, resulting in improved performance on a recently learned task. PMID- 12542674 TI - Accumbal dopamine and serotonin in anticipation of the next aggressive episode in rats. AB - Autonomic and limbic neural activities are linked to aggressive behavior, and it is hypothesized that activities in the cardiovascular and monoaminergic systems play a role in preparing for an aggressive challenge. The objective was to learn about the emergence of monoamine activity in nucleus accumbens before an aggressive confrontation that was omitted at the regular time of occurrence, dissociating the motoric from the aminergic activity. Dopamine, serotonin, heart rate and behavioral activity were monitored before, during and after a single 10 min confrontation in resident male Long-Evans rats fitted with a microdialysis probe in the n. accumbens and with a telemetry sender (experiment 1). DA, but not 5-HT efflux, was confirmed to increase in n. accumbens during and after a single aggressive episode. In aggressive males that confronted an opponent daily for 10 days (experiment 2) heart rate rose 1 h before the regularly scheduled encounter relative to control rats, as measured on day 11 in the absence of any aggression. Concurrently, DA levels increased by 60-70% over baseline levels and 5-HT levels decreased by 30-35% compared to baseline levels. These changes were sustained over 1 h, and contrasted with no significant changes in DA, 5-HT, heart rate or behavioral activity in control rats. The rise in mesolimbic DA appears to be significant in anticipating the physiological and behavioral demands of an aggressive episode, and the fall in 5-HT in its termination, dissociated from the actual execution of the behavior. PMID- 12542675 TI - Serial analysis of gene expression predicts structural differences in hippocampus of long attack latency and short attack latency mice. AB - The genetically selected long attack latency (LAL) and short attack latency (SAL) mice differ in a wide variety of behavioural traits and display differences in the serotonergic system and the hypothalamus-pituitary-adrenocortical (HPA)-axis. Serial analysis of gene expression (SAGE) was used to generate a hippocampal expression profile of almost 30 000 genes in LAL and SAL mice. Using SAGE, we found differential expression of 191 genes. Among these were genes involved in growth, signal transduction, and cell metabolism. The SAGE study was supported by GeneChip analysis (Affymetrix). Strikingly, both SAGE and GeneChips showed a higher expression of numerous cytoskeleton genes, such as cofilin and several tubulin isotypes in LAL mice. LAL mice also showed a higher expression of several calmodulin-related genes and genes encoding components of a MAPK cascade, namely raf-related oncogene and ERK2. The findings were confirmed by in situ hybridization. Our results of differential expression of cytoskeleton and signal transduction genes therefore suggest differential regulation of the raf/ERK pathway that may be related to structural differences in the hippocampus of LAL and SAL mice. As stress-related disorders, such as depression, are also linked to differential regulation of the HPA-axis and the serotonergic system and are associated with altered hippocampal morphology, differential regulation of these genes may be involved in the pathogenesis of these diseases. PMID- 12542676 TI - Age-dependent cognitive decline in the APP23 model precedes amyloid deposition. AB - Heterozygous APP23 mice, expressing human amyloid-precursor protein with the Swedish double mutation and control littermates, were subjected to behavioral and neuromotor tasks at the age of 6-8 weeks, 3 and 6 months. A hidden-platform Morris-type water maze showed an age-dependent decline of spatial memory capacities in the APP23 model. From the age of 3 months onwards, the APP23 mice displayed major learning and memory deficits as demonstrated by severely impaired learning curves during acquisition and impaired probe trial performance. In addition to the cognitive deficit, APP23 mice displayed disturbed activity patterns. Overnight cage-activity recording showed hyperactivity in the transgenics for the three age groups tested. However, a short 2-h recording during dusk phase demonstrated lower activity levels in 6-month-old APP23 mice as compared to controls. Moreover, at this age, APP23 mice differed from control littermates in exploration and activity levels in the open-field paradigm. These findings are reminiscent of disturbances in circadian rhythms and activity observed in Alzheimer patients. Determination of plaque-associated human amyloid beta 1-42 peptides in brain revealed a fivefold increase in heterozygous APP23 mice at 6 months as compared to younger transgenics. This increase coincided with the first appearance of plaques in hippocampus and neocortex. Spatial memory deficits preceded plaque formation and increase in plaque-associated amyloid-beta 1-42 peptides, but probe trial performance did correlate negatively with soluble amyloid-beta brain concentration in 3-month-old APP23 mutants. Detectable plaque formation is not the (only) causal factor contributing to memory defects in the APP23 model. PMID- 12542677 TI - Patterns of calcium-binding proteins support parallel and hierarchical organization of human auditory areas. AB - The human primary auditory cortex (AI) is surrounded by several other auditory areas, which can be identified by cyto-, myelo- and chemoarchitectonic criteria. We report here on the pattern of calcium-binding protein immunoreactivity within these areas. The supratemporal regions of four normal human brains (eight hemispheres) were processed histologically, and serial sections were stained for parvalbumin, calretinin or calbindin. Each calcium-binding protein yielded a specific pattern of labelling, which differed between auditory areas. In AI, defined as area TC [see C. von Economo and L. Horn (1930) Z. Ges. Neurol. Psychiatr.,130, 678-757], parvalbumin labelling was dark in layer IV; several parvalbumin-positive multipolar neurons were distributed in layers III and IV. Calbindin yielded dark labelling in layers I-III and V; it revealed numerous multipolar and pyramidal neurons in layers II and III. Calretinin labelling was lighter than that of parvalbumin or calbindin in AI; calretinin-positive bipolar and bitufted neurons were present in supragranular layers. In non-primary auditory areas, the intensity of labelling tended to become progressively lighter while moving away from AI, with qualitative differences between the cytoarchitectonically defined areas. In analogy to non-human primates, our results suggest differences in intrinsic organization between auditory areas that are compatible with parallel and hierarchical processing of auditory information. PMID- 12542678 TI - Protein 4.1B associates with both Caspr/paranodin and Caspr2 at paranodes and juxtaparanodes of myelinated fibres. AB - Caspr/paranodin, a neuronal transmembrane glycoprotein, is essential for the structure and function of septate-like paranodal axoglial junctions at nodes of Ranvier. A closely related protein, Caspr2, is concentrated in juxtaparanodal regions where it associates indirectly with the shaker-type potassium channels. Although ultrastructural studies indicate that paranodal complexes are linked to the cytoskeleton, the intracellular partners of Caspr/paranodin, as well as those of Caspr2, are poorly characterized. We show that the conserved intracellular juxtamembrane regions (GNP motif) of Caspr/paranodin and Caspr2 bind proteins 4.1R and 4.1B. 4.1B is known to be enriched in paranodal and juxtaparanodal regions. 4.1B immunoreactivity accumulates progressively at paranodes and juxtaparanodes during postnatal development, following the concentration of Caspr/paranodin and Caspr2, respectively, in central and peripheral myelinated axons. These two proteins coimmunoprecipitated with 4.1B in brain homogenates. Our results provide strong evidence for the association of 4.1B with Caspr/paranodin at paranodes and with Caspr2 at juxtaparanodes. We propose that 4.1B anchors these axonal proteins to the actin-based cytoskeleton in these two regions. PMID- 12542679 TI - The serotonin transporter intronic VNTR enhancer correlated with a predisposition to affective disorders has distinct regulatory elements within the domain based on the primary DNA sequence of the repeat unit. AB - We have demonstrated that a variable number tandem repeat domain (VNTR) within intron 2 of the serotonin transporter gene is a transcriptional regulatory domain which is potentially correlated with a predisposition to affective disorders and other behavioural conditions. This correlation based on copy number of the VNTR alone (nine, 10 or 12 copies of 16/17 base-pair element) has been controversial and not reproduced in all studies. We demonstrate that individual repeat elements within the VNTR domain differ in their enhancer activity in an embryonic stem cell model. This has implications for both the mechanism by which these VNTRs are correlated with the progression of the disease and suggests that clinical analysis should now be extended to correlate sequence variation within the VNTR with the disorder. The latter may resolve some of the conflicting data published to date. PMID- 12542680 TI - Attention-dependent coupling between beta activities recorded in the cat's thalamic and cortical representations of the central visual field. AB - We have previously proposed that enhanced 16-24 Hz (beta) local field potential activity in the primary visual cortex and lateral geniculate nucleus may be an electrophysiological correlate of the attentional mechanism that increases the gain of afferent visual information flow to the cortex. In this study, we measured coupling between beta signals recorded in the thalamic (i.e. lateral geniculate or perigeniculate) and cortical representations of the central visual field (within 5 degrees from area centralis), during visual and auditory attentive situations. Signal coupling was calculated in two ways: (i) by means of crosscorrelation between raw beta activities, which depends primarily on phase coherence, and (ii) by phase-independent crosscorrelation between amplitude envelopes of beta activities. Mean amplitudes of raw signal cross correlations obtained for thalamo-cortical recording pairs were not significantly different when calculated during behavioural demands for either visual or auditory attention. In contrast, amplitudes of envelope cross correlations obtained during behaviour requiring visual attention were, on average, two times higher than those calculated during the auditory task. This attention-related coupling emerged from synchronized amplitude modulation of beta oscillatory activity that occurs within the cortico-thalamic circuit involved in central vision. PMID- 12542682 TI - Dental education and registration in an expanding European Union. PMID- 12542683 TI - Providing support for problem-based learning in dentistry: the Manchester experience. AB - The introduction of problem-based learning (PBL) into any programme demands a period of adjustment on the part of faculty. Similarly, students new to PBL take time to adapt to what is, for the majority of them, an unfamiliar mode of learning. At Manchester, closed loop PBL is used throughout the first and second years of the dental programme; the method is interdisciplinary; there are no subject boundaries. Dental students work in groups of between 10 and 15, facilitated by a tutor from the Department of Biological Sciences, to research topics and share information in a mutually supportive environment. Each week a different problem forms the focus for learning. In this paper, we seek to describe the measures introduced in response to student feedback collected via routine meetings with the senior tutor, after meetings with their academic or personal tutors and through discussion at the staff students' committee, which we at Manchester have taken to facilitate the process of adaptation to PBL. Changes have been made in the areas of recruitment, pre-admission interviewing, induction (development of an induction booklet and communication skills module) and tutorial support (overhaul of personal tutor system and introduction of peer assisted study (PAS) and personal and academic development programmes (PADPs)). Feedback on these changes, gathered via the routes described above, has been positive and continues to be central to our processes of development in these areas. Although the various ways in which PBL has been implemented worldwide may place limits on the transferability of our methods, this paper serves to illustrate some of the means available to support students in the transition to self-directed learning. The latter is not only an essential component of PBL but also something we should be seeking to foster in all students, no matter what philosophy and method of course delivery are utilized. PMID- 12542684 TI - A pilot study comparing the effectiveness of conventional training and virtual reality simulation in the skills acquisition of junior dental students. AB - The use of virtual reality (VR) in the training of operative dentistry is a recent innovation and little research has been published on its efficacy compared to conventional training methods. To evaluate possible benefits, junior undergraduate dental students were randomly assigned to one of three groups: group 1 as taught by conventional means only; group 2 as trained by conventional means combined with VR repetition and reinforcement (with access to a human instructor for operative advice); and group 3 as trained by conventional means combined with VR repetition and reinforcement, but without instructor evaluation/advice, which was only supplied via the VR-associated software. At the end of the research period, all groups executed two class 1 preparations that were evaluated blindly by 'expert' trainers, under traditional criteria (outline, retention, smoothness, depth, wall angulation and cavity margin index). Analyses of resulting scores indicated a lack of significant differences between the three groups except for scores for the category of 'outline form', for group 2, which produced significantly lower (i.e. better) scores than the conventionally trained group. A statistical comparison between scores from two 'expert' examiners indicated lack of agreement, despite identical written and visual criteria being used for evaluation by both. Both examiners, however, generally showed similar trends in evaluation. An anonymous questionnaire suggested that students recognized the benefits of VR training (e.g. ready access to assessment, error identification and how they can be corrected), but the majority felt that it would not replace conventional training methods (95%), although participants recognized the potential for development of VR systems in dentistry. The most common reasons cited for the preference of conventional training were excessive critical feedback (55%), lack of personal contact (50%) and technical hardware difficulties (20%) associated with VR-based training. PMID- 12542685 TI - Instructional multimedia programs for self-directed learning in undergraduate and postgraduate training in orthodontics. AB - OBJECTIVES: The objectives of this study were to develop and evaluate an interactive multimedia courseware package in orthodontics and to provide dental undergraduate and postgraduate students with an interactive means of self-study and self-evaluation. METHODS: A commercially available software authoring system was used to develop the courseware package. The courseware was presented for evaluation through a questionnaire to four different groups of undergraduate and postgraduate students. Each group consisted of 25 students. The first three groups were third, fourth, and fifth year of dental curriculum at the Katholieke Universiteit Leuven, Belgium. The fourth group comprised postgraduate students. A one-way ANOVA was carried out in order to check for statistical differences, whereas a Scheffe's test was used in order to locate the statistical differences between groups. The p value was set at 0.05. RESULTS: The majority of undergraduate and postgraduate students in this study were very enthusiastic about this form of educational approach (74/100) because it was experienced as very helpful in understanding the orthodontic curriculum more efficiently (98/100). CONCLUSIONS: The program provides both undergraduate and postgraduate dental students with a useful interactive means of self-study and self evaluation. PMID- 12542686 TI - The use of the Internet within a dental school. AB - The Internet is an increasingly popular medium for delivering educational material. The aim of this study was to determine the attitudes of students and their clinical teachers to the use of the Internet within a dental school in the UK. Questionnaires were distributed to undergraduate dental students in the three clinical years and to all their clinical academic teachers. All students and staff have access to computers and Internet at the university. The majority (72%) of students have access to a computer and 53% also have access to the Internet at home. Of the academic staff, 91% have a computer and 68% have access to the Internet at home. The reasons for use of the Internet differed between staff and students. Whilst equal proportions of students used the Internet for dentistry (38%) and for pleasure (35%), a higher proportion of staff used the Internet more for dentistry (36%) than for pleasure (14%). Students highlighted cost and time as barriers to Internet use, whereas staff lacked confidence in their ability to use the Internet. Less than half (44%) of the students are confident in the accuracy of information from the Internet compared to almost two-thirds (64%) of staff. This study revealed differences in the attitudes of staff and students to the use of Internet as a resource for dentistry. Students are positive to the suggestion that lectures should be presented on the web. Most students (74%) did not see that this would influence attendance at lectures whilst 91% of staff stated that it would decrease lecture attendance. In conclusion, this study revealed differences in the attitudes of staff and students to the use of Internet as a resource for dentistry. PMID- 12542687 TI - Recording and understanding social histories by dental undergraduates in a community-based clinical programme. AB - Although social inequalities are known to account for large differences in dental health, attitudes to dental attendance and the type of treatment received by the patient, the taking of a patient's social history receives little emphasis in dental undergraduate teaching. Social history is defined as recording the social and family circumstances of the patient. An exercise undertaken by undergraduate dental students on clinical placements in the Community Dental Service (CDS) is described. Students write a profile of the community in which the clinic is situated, write case histories for two of their patients and then highlight issues illustrated by the case histories with reference to information on the wider community and published literature on inequalities in health and barriers to care. Students completing the exercise in April 2000 and their tutors were given additional information, including a checklist to help in the completion of the social histories of the two patients. A comparison with the reports submitted in the previous year showed that this significantly improved the recording and understanding of social history. This exercise provides a means whereby dental public health issues can be taught to undergraduates whilst maintaining relevance to the clinical setting. PMID- 12542688 TI - The repair of direct composite restorations: an international survey of the teaching of operative techniques and materials. AB - This article reports the findings of a study on the aspects of the teaching of repair as a conservative alternative to the replacement of failing direct composite restorations (DCRs) in primary dental degree curricula in the UK and Ireland, Germany and Scandinavia. Data on teaching, including operative techniques applied in the repair of DCRs, were collected by means of a questionnaire sent to 58 dental schools in the surveyed countries. Based on the overall response rate of 83%, the findings indicate that the majority (59%) of the schools in the countries surveyed may be found to teach the repair of DCRs. However, marked variations were observed in respect of this teaching and the expected longevity of repaired DCRs. Where the repair of DCRs was not taught, views differed as to the intentions, if any, to include this teaching in the curriculum. It is concluded that the teaching of DCRs may be found to be widespread in dental schools in the UK and Ireland, Germany and Scandinavia, with this teaching being subjected to considerable variation within and between countries. PMID- 12542689 TI - Assessment of telomere length and factors that contribute to its stability. AB - Short strands of tandem hexameric repeats known as telomeres cap the ends of linear chromosomes. These repeats protect chromosomes from degradation and prevent chromosomal end-joining, a phenomenon that could occur due to the end replication problem. Telomeres are maintained by the activity of the enzyme telomerase. The total number of telomeric repeats at the terminal end of a chromosome determines the telomere length, which in addition to its importance in chromosomal stabilization is a useful indicator of telomerase activity in normal and malignant tissues. Telomere length stability is one of the important factors that contribute to the proliferative capacity of many cancer cell types; therefore, the detection and estimation of telomere length is extremely important. Until relatively recently, telomere lengths were analyzed primarily using the standard Southern blot technique. However, the complexities of this technique have led to the search for more simple and rapid detection methods. Improvements such as the use of fluorescent probes and the ability to sort cells have greatly enhanced the ease and sensitivity of telomere length measurements. Recent advances, and the limitations of these techniques are evaluated. Drugs that assist in telomere shortening may contribute to tumor regression. Therefore, factors that contribute to telomere stability may influence the efficiency of the drugs that have potential in cancer therapy. These factors in relation to telomere length are also examined in this analysis. PMID- 12542690 TI - Novel aspects of calmodulin target recognition and activation. AB - Several crystal and NMR structures of calmodulin (CaM) in complex with fragments derived from CaM-regulated proteins have been reported recently and reveal novel ways for CaM to interact with its targets. This review will discuss and compare features of the interaction between CaM and its target domains derived from the plasma membrane Ca2+-pump, the Ca2+-activated K+-channel, the Ca2+/CaM-dependent kinase kinase and the anthrax exotoxin. Unexpected aspects of CaM/target interaction observed in these complexes include: (a) binding of the Ca2+-pump domain to only the C-terminal part of CaM (b) dimer formation with fragments of the K+-channel (c) insertion of CaM between two domains of the anthrax exotoxin (d) binding of Ca2+ ions to only one EF-hand pair and (e) binding of CaM in an extended conformation to some of its targets. The mode of interaction between CaM and these targets differs from binding conformations previously observed between CaM and peptides derived from myosin light chain kinase (MLCK) and CaM-dependent kinase IIalpha (CaMKIIalpha). In the latter complexes, CaM engulfs the CaM binding domain peptide with its two Ca2+-binding lobes and forms a compact, ellipsoid-like complex. In the early 1990s, a model for the activation of CaM regulated proteins was developed based on this observation and postulated activation through the displacement of an autoinhibitory or regulatory domain from the target protein upon binding of CaM. The novel structures of CaM-target complexes discussed here demonstrate that this mechanism of activation may be less general than previously believed and seems to be not valid for the anthrax exotoxin, the CaM-regulated K+-channel and possibly also not for the Ca2+-pump. PMID- 12542691 TI - Stoichiometric network theory for nonequilibrium biochemical systems. AB - We introduce the basic concepts and develop a theory for nonequilibrium steady state biochemical systems applicable to analyzing large-scale complex isothermal reaction networks. In terms of the stoichiometric matrix, we demonstrate both Kirchhoff's flux law sigma(l)J(l)=0 over a biochemical species, and potential law sigma(l) mu(l)=0 over a reaction loop. They reflect mass and energy conservation, respectively. For each reaction, its steady-state flux J can be decomposed into forward and backward one-way fluxes J = J+ - J-, with chemical potential difference deltamu = RT ln(J-/J+). The product -Jdeltamu gives the isothermal heat dissipation rate, which is necessarily non-negative according to the second law of thermodynamics. The stoichiometric network theory (SNT) embodies all of the relevant fundamental physics. Knowing J and deltamu of a biochemical reaction, a conductance can be computed which directly reflects the level of gene expression for the particular enzyme. For sufficiently small flux a linear relationship between J and deltamu can be established as the linear flux-force relation in irreversible thermodynamics, analogous to Ohm's law in electrical circuits. PMID- 12542692 TI - Mosquito (Aedes aegypti ) aquaporin, present in tracheolar cells, transports water, not glycerol, and forms orthogonal arrays in Xenopus oocyte membranes. AB - Previous results showed that mRNA encoding a putative aquaporin (AQP) (GenBank accession number AF218314) is present in the tracheolar cells associated with female Aedes aegypti Malpighian tubules. In this study, immunohistochemistry detected the protein, AeaAQP, also in tracheolar cells, suggesting its involvement in water movement in the respiratory system. When expressed in Xenopus oocytes, AeaAQP increased the osmotic water permeability from 15 x 10(-6) to 150 x 10(-6) m x s-1, which was inhibited by mercury ions. No permeability to glycerol or other solute was observed. AeaAQP expressed in oocytes was solubilized as a homotetramer in nondenaturing detergent as deduced from velocity centrifugation on density gradients. Phylogenetic analysis of MIP (major intrinsic protein) family sequences shows that AeaAQP clusters with other native orthogonal array forming proteins. Specific orthogonal arrays were detected by freeze-fracture analysis of AeaAQP oocyte membranes. We conclude that, in tracheolar cells of A. aegypti, AeaAQP is probably a highly water-permeable homotetrameric MIP which natively can form 2D crystals. PMID- 12542693 TI - Elementary modes analysis of photosynthate metabolism in the chloroplast stroma. AB - We briefly review the metabolism of the chloroplast stroma, and describe the structural modelling technique of elementary modes analysis. The technique is applied to a model of chloroplast metabolism to investigate viable pathways in the light, in the dark, and in the dark with the addition of sedoheptulose-1,7 bisphosphatase (normally inactive in the dark). The results of the analysis show that it is possible for starch degradation to enhance photosynthetic triose phosphate export in the light, but the reactions of the Calvin cycle alone are not capable of providing a sustainable flux from starch to triose phosphate in the dark. When reactions of the oxidative pentose phosphate pathway are taken into consideration, triose phosphate export in the dark becomes possible by the operation of a cyclic pathway not previously described. The effect of introducing sedoheptulose-1,7-bisphosphatase to the system are relatively minor and, we predict, innocuous in vivo. We conclude that, in contrast with the traditional view of the Calvin cycle and oxidative pentose phosphate pathway as separate systems, they are in fact, in the context of the chloroplast, complementary and overlapping components of the same system. PMID- 12542694 TI - Endotoxic activity and chemical structure of lipopolysaccharides from Chlamydia trachomatis serotypes E and L2 and Chlamydophila psittaci 6BC. AB - The lipopolysaccharide (LPS) of Chlamydia trachomatis serotype E was isolated from tissue culture-grown elementary bodies and analyzed structurally by mass spectrometry and 1H, 13C and 31P nuclear magnetic resonance. The LPS is composed of the same pentasaccharide bisphosphate alphaKdo-(2-8)-alphaKdo-(2-4)-alphaKdo (2-6)-betaGlcN-4P-(1-6)-alphaGlcN-1P (Kdo is 3-deoxy-alpha-d-manno-oct-2-ulosonic acid) as reported for C. trachomatis serotype L2[Rund, S., Lindner, B., Brade, H. and Holst, O. (1999) J. Biol. Chem. 274, 16819-16824]. The glucosamine disaccharide backbone is substituted with a complex mixture of fatty acids with ester or amide linkage whereby no ester-linked hydroxy fatty acids were found. The LPS was purified carefully (with contaminations by protein or nucleic acids below 0.3%) and tested for its ability to induce proinflammatory cytokines in several readout systems in comparison to LPS from C. trachomatis serotype L2 and Chlamydophila psittaci strain 6BC as well as enterobacterial smooth and rough LPS and synthetic hexaacyl lipid A. The chlamydial LPS were at least 10 times less active than typical endotoxins; specificity of the activities was confirmed by inhibition with the LPS antagonist, B1233, or with monoclonal antibodies against chlamydial LPS. Like other LPS, the chlamydial LPS used toll-like receptor TLR4 for signalling, but unlike other LPS activation was strictly CD14-dependent. PMID- 12542695 TI - Trophoblast-like human choriocarcinoma cells serve as a suitable in vitro model for selective cholesteryl ester uptake from high density lipoproteins. AB - As human choriocarcinoma cells display many of the biochemical and morphological characteristics reported for in utero invasive trophoblast cells we have studied cholesterol supply from high density lipoproteins (HDL) to these cells. Binding properties of 125I-labeled HDL subclass 3 (HDL3) at 4 degrees C were similar for BeWo, JAr, and Jeg3 choriocarcinoma cell lines while degradation rates at 37 degrees C were highest for BeWo. Calculating the selective cholesteryl ester (CE) uptake as the difference between specific cell association of [3H]CE-labeled HDL3 and holoparticle association of 125I-labeled HDL3 revealed that in BeWo cells, the selective CE-uptake was slightly lower than holoparticle association. However, the pronounced capacity for specific cell association of [3H]CE-HDL3 and selective [3H]CE-uptake in excess of HDL3-holoparticle association, and cAMP mediated enhanced cell association of [3H]CE-HDL3 in JAr and Jeg3 suggested the scavenger receptor class B, type I (SR-BI) to be responsible for this pathway. Abundant expression of SR-BI (but not SR-BII, a splice variant of SR-BI) could be observed in JAr and Jeg3 but not in BeWo cells using RT-PCR, Northern and Western blot analysis, and immunocytochemical technique. Adenovirus-mediated overexpression of SR-BI in all three choriocarcinoma cell lines resulted in an enhanced capacity for cell association of [3H]CE-HDL3 (20-fold in BeWo; fivefold in JAr and Jeg3). The fact that exogenous HDL3 remarkably increases proliferation in JAr and Jeg3 supports the notion that selective CE-uptake and subsequent intracellular generation of cholesterol is coupled to cellular growth. From our findings we propose that JAr and Jeg3 cells serve as a suitable in vitro model to study selective CE-supply to human placental cells. PMID- 12542696 TI - tmRNA from Thermus thermophilus. Interaction with alanyl-tRNA synthetase and elongation factor Tu. AB - The interaction of a Thermus thermophilus tmRNA transcript with alanyl-tRNA synthetase and elongation factor Tu has been studied. The synthetic tmRNA was found to be stable up to 70 degrees C. The thermal optimum of tmRNA alanylation was determined to be around 50 degrees C. At 50 degrees C, tmRNA transcript was aminoacylated by alanyl-tRNA synthetase with 5.9 times lower efficiency (kcat/Km) than tRNAAla, primarily because of the difference in turnover numbers (kcat). Studies on EF-Tu protection of Ala approximately tmRNA against alkaline hydrolysis revealed the existence of at least two different binding sites for EF Tu on charged tmRNA. The possible nature of these binding sites is discussed. PMID- 12542697 TI - New insights into the P-glycoprotein-mediated effluxes of rhodamines. AB - Multidrug resistance (MDR) in tumour cells is often caused by the overexpression of the plasma drug transporter P-glycoprotein (P-gp). This protein is an active efflux pump for chemotherapeutic drugs, natural products and hydrophobic peptides. Despite the advances of recent years, we still have an unclear view of the molecular mechanism by which P-gp transports such a wide diversity of compounds across the membrane. Measurement of the kinetic characteristics of substrate transport is a powerful approach to enhancing our understanding of their function and mechanism. The aim of the present study was to further characterize the transport of several rhodamine analogues, either positively charged or zwitterionic. We took advantage of the intrinsic fluorescence of rhodamines and performed a flow-cytometric analysis of dye accumulation in the wild-type drug sensitive K562 that do not express P-gp and its MDR subline that display high levels of MDR. The measurements were made in real time using intact cells. The kinetic parameter, ka = VM/km, which is a measure of the efficiency of the P-gp-mediated efflux of a substrate was similar for almost all the rhodamine analogues tested. In addition these values were compared with those determined previously for the P-gp-mediated efflux of anthracycline. Our conclusion is that the compounds of these two classes of molecules, anthracyclines and rhodamines, are substrates of P-gp and that their pumping rates at limiting low substrate concentration are similar. The findings presented here are the first to show quantitative information about the kinetic parameters for P-gp-mediated efflux of rhodamine analogues in intact cells. PMID- 12542698 TI - Role of Acinetobacter calcoaceticus 3,4-dihydrocoumarin hydrolase in oxidative stress defence against peroxoacids. AB - The physiological role of a bifunctional enzyme, 3,4-dihydrocoumarin hydrolase (DCH), which is capable of both hydrolysis of ester bonds and organic acid assisted bromination of organic compounds, was investigated. Purified DCH from Acinetobacter calcoaceticus F46 catalysed dose- and time-dependent degradation of peracetic acid. The gene (dch) was cloned from the chromosomal DNA of the bacterium. The dch ORF was 831 bp long, corresponding to a protein of 272 amino acid residues, and the deduced amino acid sequence showed high similarity to those of bacterial serine esterases and perhydrolases. The dch gene was disrupted by homologous recombination on the A. calcoaceticus genome. The dch disruptant strain was more sensitive to growth inhibition by peracetic acid than the parent strain. On the other hand, the recombinant Escherichia coli cells expressing dch were more resistant to peracetic acid. A putative catalase gene was found immediately downstream of dch, and Northern blot hybridization analysis revealed that they are transcribed as part of a polycistronic mRNA. These results suggested that in vivo DCH detoxifies peroxoacids in conjunction with the catalase, i.e. peroxoacids are first hydrolysed to the corresponding acids and hydrogen peroxide by DCH, and then the resulting hydrogen peroxide is degraded by the catalase. PMID- 12542699 TI - Selecting cells with different Alzheimer's disease gamma-secretase activity using FACS. Differential effect on presenilin exon 9 gamma- and epsilon-cleavage. AB - The ultimate step in Alzheimer's disease Abeta generation involves gamma secretase, which releases Abeta from its membrane-bound precursor. A similar presenilin-dependent proteolytic activity is implicated in the release of the Notch intracellular domain. We have developed a novel assay for gamma-secretase activity based on green fluorescent protein detection. This involves cotransfection of a substrate-activator based on the amyloid precursor protein or the Notch sequence and a fluorescent reporter gene. Stable fluorescent cell populations were selected by fluorescent activated cell sorting and characterized. This assay enabled the identification and sorting of populations, which differ in their levels of gamma-secretase activity, with high fluorescent cells producing more Abeta than low fluorescent cells. Specific gamma-secretase inhibitors, L-685,458 and MW167, reduced cell fluorescence in a dose-dependent manner that paralleled inhibition of Abeta secretion. Overexpression of presenilin 1 increased the cell fluorescence. Cells expressing presenilin with different aspartate mutations (D257A, D385A and D257A/D385A) or exon 9 deletion mutation showed reduced fluorescence. The single aspartate mutations showed a concomitant reduction in Abeta secretion, whereas the D257A/D385A and DeltaE9 mutations had no effect on Abeta secretion. PMID- 12542700 TI - Bovine tryptases. cDNA cloning, tissue specific expression and characterization of the lung isoform. AB - A complementary DNA encoding a new bovine tryptase isoform (here named BLT) was cloned and sequenced from lung tissue. Analysis of sequence indicates the presence of a 26-amino acid prepro-sequence and a 245 amino acid catalytic domain. It contains six different residues when compared with the previously characterized tryptase from bovine liver capsule (BLCT), with the most significant difference residing at the primary specificity S1 pocket. In BLT, the canonical residues Asp-Ser are present at positions 188-189, while in BLCT these positions are occupied by residues Asn-Phe. This finding was confirmed by mass fingerprinting of the peptide mixture obtained upon in-gel tryptic digestion of BLT. Analysis by gel filtration of the purified protein shows that BLT is probably tetrameric, similar to the previously identified tryptases from other species, with monomer migrating as 35-40 kDa multiple bands in SDS/PAGE. As expected, the catalytic abilities of the two bovine tryptases are different. The specificity constant values (kcat/Km) assayed with model substrates are 10- to 60 fold higher in the case of BLT. The tissue-specific expression of the two tryptases was evaluated at the RNA level by analysis of their different restriction patterns. In lung, only BLT was found to be expressed, while in liver capsule only BLCT is present. Both isoforms are distributed in similar amounts in heart and spleen. Analysis of the two gene sequences reveals the presence of several recognition sequences in the promoter regions and suggest a role for hormones in governing the mechanism of tissue expression of bovine tryptases. PMID- 12542701 TI - The transthyretin-related protein family. AB - A number of proteins related to the homotetrameric transport protein transthyretin (TTR) forms a highly conserved protein family, which we present in an integrated analysis of data from different sources combined with an initial biochemical characterization. Homologues of the transthyretin-related protein (TRP) can be found in a wide range of species including bacteria, plants and animals, whereas transthyretins have so far only been identified in vertebrates. A multiple sequence alignment of 49 TRP sequences from 47 species to TTR suggests that the tertiary and quaternary features of the three-dimensional structure are most likely preserved. Interestingly, while some of the TRP orthologues show as little as 30% identity, the residues at the putative ligand-binding site are almost entirely conserved. RT/PCR analysis in Caenorhabditis elegans confirms that one TRP gene is transcribed, spliced and predominantly expressed in the worm, which suggests that at least one of the two C. elegans TRP genes encodes a functional protein. We used double-stranded RNA-mediated interference techniques in order to determine the loss-of-function phenotype for the two TRP genes in C. elegans but detected no apparent phenotype. The cloning and initial characterization of purified TRP from Escherichia coli reveals that, while still forming a homotetramer, this protein does not recognize thyroid hormones that are the natural ligands of TTR. The ligand for TRP is not known; however, genomic data support a functional role involving purine catabolism especially linked to urate oxidase (uricase) activity. PMID- 12542702 TI - Probing suggested catalytic domains of glycosyltransferases by site-directed mutagenesis. AB - The plant enzyme arbutin synthase isolated from cell suspension cultures of Rauvolfia serpentina and heterologously expressed in Escherichia coli is a member of the NRD1beta family of glycosyltransferases. This enzyme was used to prove, by site-directed mutagenesis, suggested catalytic domains and reaction mechanisms proposed for enzyme-catalyzed glycosylation. Replacement of amino acids far from the NRD domain do not significantly affect arbutin synthase activity. Exchange of amino acids at the NRD site leads to a decrease of enzymatic activity, e.g. substitution of Glu368 by Asp. Glu368, which is a conserved amino acid in glycosyltransferases located at position 2 and is important for enzyme activity, does not serve as the nucleophile in the catalytic centre as proposed. When it is replaced by Ala, the resulting mutant enzyme E368A exhibits comparable activity as found for E368D in respect to vanillin. Enzyme activities of wild-type and E368A towards several substrates were not affected at the same level. His360 at position 1 of NRD1beta glycosyltransferases occupies a more crucial role as expected. When it is exchanged against other basic amino acids such as Lys or Arg the enzyme activity decreases approximately 1000-fold. Replacement of His360 by Glu leads to a mutant enzyme (H360E) with an approximately 4000-fold lower activity compared with the wild-type. This mutein still produces a beta glucoside, not an alpha-glucoside and therefore indicates that generation of the typical E-E motif of NRD1alpha glycosyltransferases does not convert a NRD1beta enzyme into a NRD1alpha enzyme. The presented data do not support several suggestions made in the literature about catalytic amino acids involved in the glycosyltransfer reaction. PMID- 12542703 TI - Cofactor-independent oxygenation reactions catalyzed by soluble methane monooxygenase at the surface of a modified gold electrode. AB - Soluble methane monooxygenase (sMMO) is a three-component enzyme that catalyses dioxygen- and NAD(P)H-dependent oxygenation of methane and numerous other substrates. Oxygenation occurs at the binuclear iron active centre in the hydroxylase component (MMOH), to which electrons are passed from NAD(P)H via the reductase component (MMOR), along a pathway that is facilitated and controlled by the third component, protein B (MMOB). We previously demonstrated that electrons could be passed to MMOH from a hexapeptide-modified gold electrode and thus cyclic voltammetry could be used to measure the redox potentials of the MMOH active site. Here we have shown that the reduction current is enhanced by the presence of catalase or if the reaction is performed in a flow-cell, probably because oxygen is reduced to hydrogen peroxide, by MMOH at the electrode surface and the hydrogen peroxide then inactivates the enzyme unless removed by catalase or a continuous flow of solution. Hydrogen peroxide production appears to be inhibited by MMOB, suggesting that MMOB is controlling the flow of electrons to MMOH as it does in the presence of MMOR and NAD(P)H. Most importantly, in the presence of MMOB and catalase, the electrode-associated MMOH oxygenates acetonitrile to cyanoaldehyde and methane to methanol. Thus the electochemically driven sMMO showed the same catalytic activity and regulation by MMOB as the natural NAD(P)H-driven reaction and may have the potential for development into an economic, NAD(P)H-independent oxygenation catalyst. The significance of the production of hydrogen peroxide, which is not usually observed with the NAD(P)H driven system, is also discussed. PMID- 12542704 TI - Functional properties of the sex-hormone-binding globulin (SHBG)-like domain of the anticoagulant protein S. AB - Protein S (PS) possesses a sex-hormone-binding globulin (SHBG)-like domain in place of the serine-protease domain found in other vitamin K-dependent plasma proteins. This SHBG-like domain is able to bind a complement fraction, C4b binding protein (C4b-BP). To establish whether the PS SHBG-like domain can fold normally in the absence of other domains, and to obtain information on the specific functions of this region, we expressed the PS SHBG-like domain alone or together with its adjacent domain EGF4. The folding of the two recombinant modules was studied by analyzing their binding to C4b-BP. The apparent dissociation constants of this interaction indicated that both recombinant modules adopted the conformation of native PS, indicating that the PS SHBG-like region is an independent folding unit. We also obtained the first direct evidence that the SHBG-like domain alone is sufficient to support the interaction with C4b BP. In addition, both recombinant modules were able to bind Ca2+ directly, as shown by the migration shift in agarose gel electrophoresis in the presence of Ca2+, together with the results of equilibrium dialysis and the functional effect of Ca2+ on the C4b-BP/PS interaction, confirming the presence of one Ca2+ binding site within the SHBG-like domain. Neither recombinant module exhibited activated protein C (aPC) cofactor activity in a clotting assay, suggesting that the PS SHBG-like region must be part of the intact molecule for it to contribute to aPC cofactor activity, possibly by constraining the different domains in a conformation that permits optimal interaction with aPC. PMID- 12542705 TI - Two CCAAT/enhancer binding protein sites in the cytochrome P4503A1 locus. Potential role in the glucocorticoid response. AB - Induction of CYP3A genes by the ligand-activated pregnane-X-receptor (PXR) involves the interaction of other as yet unidentified liver transcription factors. Here we show that the CYP3A1 promoter contains two active sites controlled by the CCAAT/enhancer-binding protein alpha (C/EBPalpha), previously shown to regulate a number of liver stress response genes. We have identified two functional C/EBP binding sites at the CYP3A1 promoter that confer luciferase activity to C/EBPalpha cotransfected CHO cells. When inserted upstream of a thymidine kinase promoter, oligonucleotides corresponding to these elements ( 350/-311 and -628/-608), increase reporter gene expression when cotransfected with a C/EBPalpha expression vector. Point mutations in the most conserved nucleotides in either element prevent binding of C/EBPalpha and abolish transactivation of the CYP3A1 promoter. Moreover, we demonstrate that C/EBPalpha accumulates in the rat liver nuclei in response to dexamethasone, and that under these conditions C/EBPalpha binds to the CYP3A1 promoter elements. Our results suggest a correlation between transcription of C/EBPalpha, nuclear protein function and induction of CYP3A1 by dexamethasone in the liver. They also support the notion that C/EBPalpha participates in the up-regulation of the CYP3A1 gene in response to synthetic glucocorticoids. PMID- 12542708 TI - Adaptation to nickel spiking of bacterial communities in neocaledonian soils. AB - Adaptation to nickel of bacterial communities of two extreme neocaledonian soils (an ultramafic soil and an acidic soil) was investigated by nickel spiking and compared with adaptation in a non-neocaledonian soil used as reference. Soil microcosms were amended with nickel chloride (NiCl2), and bacterial community structure was analysed with the ribosomal intergenic spacer analysis (RISA) technique. Then, bacterial populations that respond to nickel stress were identified by cloning and sequencing. In the ultramafic soil, a shift occurred on day zero on the assay profiles and consisted of the emergence of a bacterial group closely related to the Ralstonia/Oxalobacter/Burkholderia group. It is hypothesized that NiCl2 had a physico-chemical impact on soil structure. Fourteen days after nickel spiking, another shift occurred in the two soils that concerned a bacterial group belonging to the Actinomycete group. Only a few changes occurred in the bacterial community structure of the neocaledonian soils compared with those of the reference soil, which is more affected by nickel spiking. These results suggest that neocaledonian soil bacteria are particularly well adapted to nickel. PMID- 12542709 TI - Molecular diversity and characterization of nitrite reductase gene fragments (nirK and nirS) from nitrate- and uranium-contaminated groundwater. AB - Nitrate-contaminated groundwater samples were analysed for nirK and nirS gene diversity. The samples differed with respect to nitrate, uranium, heavy metals, organic carbon content, pH and dissolved oxygen levels. A total of 958 nirK and 1162 nirS clones were screened by restriction fragment length polymorphism (RFLP) analysis: 48 and 143 distinct nirK and nirS clones, respectively, were obtained. A single dominant nirK restriction pattern was observed for all six samples and was 83% identical to the Hyphomicrobium zavarzinii nirK gene. A dominant nirS pattern was observed for four of the samples, including the background sample, and was 95% identical to the nirS of Alcaligenes faecalis. Diversity indices for nirK and nirS sequences were not related to any single geochemical characteristic, but results suggested that the diversity of nirK genes was inversely proportional to the diversity of nirS. Principal component analysis (PCA) of the sites based on geochemistry grouped the samples by low, moderate and high nitrate but PCA of the unique operational taxonomic units (OTUs) distributions grouped the samples differently. Many of the sequences were not closely related to previously observed genes and some phylogenetically related sequences were obtained from similar samples. The results indicated that the contaminated groundwater contained novel nirK and nirS sequences, functional diversity of both genes changed in relation to the contaminant gradient, but the nirK and nirS functional diversity was affected differently. PMID- 12542711 TI - Potential bias of fungal 18S rDNA and internal transcribed spacer polymerase chain reaction primers for estimating fungal biodiversity in soil. AB - Four fungal 18S rDNA and internal transcribed spacer (ITS) polymerase chain reaction (PCR) primer pairs were tested for their specificity towards target fungal DNA in soil DNA extracts, and their ability to assess the diversity of fungal communities in a natural grassland soil was compared. Amplified PCR products were cloned, and approximately 50 clones from each library were sequenced. Phylogenetic analysis and database searches indicated that each of the sequenced cloned DNA fragments was of fungal origin for each primer pair, with the exception of the sequences generated using the 18S rDNA primers nu-SSU-0817 and nu-SSU-1196, where 35 of the 50 sequenced clones represented soil invertebrates. Although some of the primers have previously been suggested to be biased towards certain fungal taxonomic groups, the ratio of sequences representing each of the four main fungal phyla, Ascomycota, Basidiomycota, Chytridiomycota and Zygomycota, was similar for each of the primer pairs, suggesting that primer bias may be less significant than previously thought. Collector's curves were plotted to estimate the coverage obtained for each of the clone libraries after clustering the sequences into operational taxonomic units at a level of 99% sequence similarity. The curves indicated that good coverage of diversity was achieved, with the exception of the clone library constructed using primers nu-SSU-0817 and nu-SSU-1196, on account of the high number of non-fungal sequences obtained. The work demonstrates the usefulness of 18S rDNA and ITS PCR primers for assessing fungal diversity in environmental samples, and it also highlights some potential limitations of the approach with respect to PCR primer specificity and bias. PMID- 12542712 TI - Diversity of alpha-halocarboxylic acid dehalogenases in bacteria isolated from a pristine soil after enrichment and selection on the herbicide 2,2 dichloropropionic acid (Dalapon). AB - Five pure cultures of bacteria (strains DA1-5) able to degrade 2,2 dichloropropionic acid (22DCPA) were isolated for the first time from pristine bulk soil samples. From 16S rDNA analysis, it was concluded that strains DA2, DA3 and DA4 were members of the Bradyrhizobium subgroup (alpha-Proteobacteria), strain DA5 clustered in the Brucella assemblage (alpha-Proteobacteria) and strain DA1 clustered in the beta-Proteobacteria. Biochemical and molecular analysis of the dehalogenases from the isolates showed that these enzymes were quite diverse. Several dehalogenases were closely related to group I and II alpha-halocarboxylic acid dehalogenases, and partial polymerase chain reaction (PCR) products were obtained from isolates DA1, 2, 3 and 4 using degenerate dehalogenase primers. However, no PCR products were obtained from isolate DA5 using either of the group I or II alpha-halocarboxylic acid dehalogenase primers. Isolates DA2 and DA4 contained putative silent dehalogenases. The investigation highlighted the endemic nature of these genes in pristine environments and how diverse these were even from spatially close samples. PMID- 12542710 TI - Isolation and phylogenetic characterization of bacteria capable of inducing differentiation in the green alga Monostroma oxyspermum. AB - Many green algae cannot develop normally when they are grown under axenic conditions. Monostroma oxyspermum, for example, proliferates unicellularly in an aseptic culture, but develops into a normal foliaceous gametophyte in the presence of some marine bacteria. More than 1000 bacterial strains were isolated from marine algae and sponges and assayed for their ability to induce the morphogenesis of unicellular M. oxyspermum. Fifty bacterial strains exhibiting morphogenesis-inducing activity against unicellular M. oxyspermum were isolated. The partial gyrB (approximately 1.2 kbp) and 16S rDNA (approximately 1.4 kbp) sequences of about 40 active strains were determined, and their phylogenetic relationships were analysed. All these strains were located within the Cytophaga Flavobacterium-Bacteroides (CFB) complex, and most of these strains were clustered in a clade comprising Zobellia uliginosa. On the other hand, these bacteria also exhibited morphogenetic activity against germ-free spores of Ulva pertusa, Ulva conglobata and Enteromorpha intestinalis. Moreover, these bacteria induced the release of spores from the leafy young gametophyte of M. oxyspermum. These results indicate that strains belonging to several groups in the CFB complex play an important role in the normal development of green algae in the marine coastal environment. PMID- 12542713 TI - Diversity of microbial communities correlated to physiochemical parameters in a digestion basin of a zero-discharge mariculture system. AB - Bacterial community structure and physiochemical parameters were examined in a sedimentation basin of a zero-discharge mariculture system. The system consisted of an intensively stocked fish basin from which water was recirculated through two separate treatment loops. Surface water from the basin was pumped over a trickling filter in one loop while bottom-water was recirculated through a sedimentation basin followed by a fluidized bed reactor in the other. Ammonia oxidation to nitrate in the trickling filter and organic matter digestion together with nitrate reduction in the sedimentation basin and fluidized bed reactor, allowed zero-discharge operation of the system. Relatively high concentrations of oxygen, nitrate, sulphate and organic matter detected simultaneously in the digestion basin suggested the potential for a wide range of microbially-mediated transformation processes. In this study, physiochemical parameters were correlated to bacterial diversity and distribution in horizontal and vertical profiles within this basin in an effort to obtain a basic understanding of the chemical and microbial processes in this system. Chemical activity and microbial diversity, the latter measured by denaturing gradient gel electrophoresis (DGGE) analysis of polymerase chain reaction (PCR) amplified 16S rDNA fragments, were higher in the sludge layer than in the overlying aqueous layer of the basin. Chemical parameters in sludge samples close to the basin inlet suggested enhanced microbial activity relative to other sampling areas with evidence of both nitrate and sulphate reduction. Four of the nine DGGE bands identified in this zone were affiliated with the Bacteroidetes phylum. Detected sequences closely related to sequences of organisms involved in the sulphur cycle included Desulfovibrio, Dethiosulfovibrio and apparent sulphur oxidizers from the gamma-proteobacteria. In addition, a number of sequences from the beta and alpha proteobacteria were identified. PMID- 12542714 TI - Ecology of mixed biofilms subjected daily to a chlorinated alkaline solution: spatial distribution of bacterial species suggests a protective effect of one species to another. AB - Three bacterial strains (Kocuria sp. C714.1, Brevibacterium linens B337.1 and Staphylococcus sciuri CCL101) were grown together on stainless steel and were subjected daily to a commercial alkaline chlorine solution (22 mg l-1 of free chlorine, pH 11) over a period of 4 weeks. After the daily chemical shock, culture madia [1:20 dilution of tryptic soy broth (TSB-YE/20) or diluted whey] was deposited on the biofilms. The chemical shocks led first to a drop in the culturable population, followed by an increase and finally stabilization at around 106-107 CFU cm-2 by day 11 of the experiment. These changes in the microbial population can be attributed to a decreasing susceptibility to the antimicrobial agent with biofilm age, and to the consumption of free chlorine by biofilm exoproteins. The microbial composition appeared to be linked to the free chlorine concentration that depended on exoprotein production. At the end of the experiment, exoprotein production was greater for biofilms grown in TSBYE/20 than in whey. As a consequence, biofilms grown in whey did not neutralize the chlorine and the dominant strain was the one having the highest resistance to chlorine: K. varians. When biofilm were grown in TSBYE/20, chlorine was neutralized and the dominant strain was the one having the highest growth rate: S. sciuri. The presence of chlorine may also explain the distribution of S. sciuri cells as a ring around Kocuria sp. microcolonies. When chlorine was totally consumed by the biofilm during the chemical shock, S. sciuri was no longer grouped around Kocuria sp. microcolonies but was evenly scattered over the substratum as single cells or in small clusters, as it was before any chemical treatment. These findings strongly suggest protection of S. sciuri by Kocuria sp. microcolonies against the chlorinated solution. This phenomenon, added to the low susceptibility phenotype of the biofilm cells, could at least partly explain the survival of microbial cells in an adverse environment. PMID- 12542715 TI - Monitoring the colonization of monuments by bacteria: cultivation versus molecular methods. AB - Building materials commonly used in wall paintings and monuments (mortar, limestone and sandstone) were inoculated with an artificial consortium composed of 14 microorganisms and incubated for 6 months at 28 degrees C. The colonization of the different materials by the consortium was investigated. Culture independent techniques revealed the presence of a diversity of bacteria, whereas culture-dependent techniques yielded mainly spore-forming bacteria. The data suggest that plating leads to an overestimation of the number of spore-forming bacteria with respect to quiescent vegetative forms; the latter are less easily cultured, but are readily detected by culture-independent techniques. PMID- 12542719 TI - Researching new treatments for obesity: from neuroscience to inflammation. PMID- 12542720 TI - Human insulin resistance: the role of glucocorticoids. PMID- 12542721 TI - Evidence-based treatment of hypertension in patients with diabetes mellitus. PMID- 12542722 TI - Brain-derived neurotrophic factor modulates glucagon secretion from pancreatic alpha cells: its contribution to glucose metabolism. AB - AIM: Brain-derived neurotrophic factor (BDNF) reduces plasma glucose levels in obese db/db diabetic mice and is speculated to produce its effects via the hypothalamus, the regulatory centre of satiety and the autonomic nervous system. The potential effect of BDNF directly on peripheral endocrine organs, however, remains to be clarified. Here we report the effects of BDNF on hormonal secretion from pancreatic islets of Langerhans using their isolated culture. METHODS AND RESULTS: In an immunohistochemical study, mouse pancreatic alpha cells were stained specifically with the anti-TrkB (a specific receptor for BDNF) antibody. After 7 days culture of isolated islets of the normal mouse pancreas, 10 ng/ml BDNF decreased the secretion of glucagon per 6 h significantly less than that of the control (p = 0.016). In contrast, there were no significant changes in insulin secretion or glucagon and insulin contents in the islets cultured under the same conditions. In vivo administration of BDNF (10 mg/kg/day) to normal mice for 7 days significantly decreased their food consumption (p < 0.05). The fasting plasma glucose levels were decreased on day 7 compared with day 1 more significantly in BDNF-treated mice (p = 0.043) than in pair-fed control mice (p = 0.14). In newborn BDNF-knockout mice, fasting plasma glucose levels increased in the order of homozygote, heterozygote and wild type (p = 0.033). No apparent immunohistochemical abnormality was observed for pancreatic glucagon in the BDNF knockout mice. CONCLUSION: These data suggest that BDNF affects glucose metabolism not only with its anorectic effect but also with modulated glucagon secretion from pancreatic alpha cells. PMID- 12542723 TI - Influence of treatment with acarbose or glibenclamide on insulin sensitivity in type 2 diabetic patients. AB - AIM: The aim of our double-blind, placebo-controlled study was to compare the effect of acarbose and glibenclamide on the insulin sensitivity in type 2 diabetes. METHODS: We investigated 77 patients (mean age 58.7 years, mean BMI 27.3 kg/m2), treated by diet alone for at least 4 weeks. The subjects were randomized into three treatment groups for 16 weeks: 100 mg t.i.d. acarbose (n = 25) or 1 mg t.i.d. glibenclamide (n = 27) or one t.i.d. placebo (n = 25). Before and after therapy, the levels of fasting plasma glucose, glycosylated haemoglobin, fasting insulin, plasma glucose and insulin 1 h after a standardized breakfast were measured and insulin sensitivity determined by euglycaemic hyperinsulinaemic clamp test. RESULTS: After the treatment period, BMI in the acarbose and placebo group decreased significantly, whereas in the glibenclamide group a significant increase was observed. Fasting plasma glucose was only significant reduced under glibenclamide. The postprandial glucose decreased significantly after acarbose (13.8 vs. 11.4 mmol/l, p < 0.05) and glibenclamide treatment (14.6 vs. 11.4 mmol/l, p < 0.05) and was unchanged under placebo (13.8 vs. 13.7 mmol/l). The fasting insulin levels remained unchanged in all three groups, whereas postprandial insulin values increased significantly under glibenclamide. Neither acarbose nor glibenclamide significantly changed insulin sensitivity [acarbose: glucose disposal rate before treatment 2.3 mg/kg body weight/min/insulin, after treatment 3.2; glibenclamide 2.2 vs. 2.1; placebo 2.6 vs. 3.0]. CONCLUSIONS: Our results show a more substantial improvement of glucose control under glibenclamide than under acarbose which, however, was not associated with an increase of insulin sensitivity. PMID- 12542724 TI - Rosiglitazone (PPARgamma-agonist) attenuates atherogenesis with no effect on hyperglycaemia in a combined diabetes-atherosclerosis mouse model. AB - BACKGROUND: The administration of peroxisome proliferator-activated receptor gamma (PPARgamma) agonists to low-density lipoprotein (LDL)-receptor-deficient mice resulted in a reduction in the atherosclerotic lesion area in male mice, but not in female mice. The male mice also exhibited reduction in insulin resistance while the female mice did not. To further examine the relationship between PPARgamma agonists, insulin resistance and atherosclerosis, we used the model of accelerated atherosclerosis in male apolipoprotein E (apoE)-deficient mice rendered diabetic by low-dose streptozotocin (STZ). METHODS: Male, apoE-deficient mice (n = 48) were randomly divided into four groups. To induce diabetes, two groups received low-dose STZ and two groups served as controls. After diabetes induction, rosiglitazone (a PPARgamma agonist) was administered by oral gavage to one of the diabetic and one of the non-diabetic groups. RESULTS: Rosiglitazone reduced significantly the atherosclerotic aortic plaque area in both diabetic and non-diabetic apoE-deficient mice: 340 +/- 54 vs. 201 +/- 27 micromol2 (p = 0.001) in diabetic mice; 243 +/- 22 vs. 158 +/- 27 micromol2 (p = 0.001) in non-diabetic mice. Also, rosiglitazone reduced the correlation coefficient between plasma glucose and the degree of atherosclerosis (p < 0.0025) without affecting plasma glucose levels. The rosiglitazone-treated mice, both diabetic and non-diabetic, had higher lipid levels. CONCLUSIONS: Rosiglitazone-treated animals showed less atherosclerosis despite higher lipid levels and similar glucose levels. These data suggest a direct anti-atherogenic effect of rosiglitazone on the arterial wall. PMID- 12542725 TI - Aldose reductase gene polymorphism is associated with progression of diabetic nephropathy in Japanese patients with type 1 diabetes mellitus. AB - AIM: The objective of this study was to investigate cross-sectionally and longitudinally whether polymorphism of the (A-C)n dinucleotide repeat sequence of the aldose reductase (AR) gene may modulate risk for diabetic nephropathy or retinopathy in Japanese patients with type 1 diabetes. METHODS: We obtained DNA samples from 101 patients followed up after the onset of type 1 diabetes and analysed a (A-C)n dinucleotide repeat polymorphic marker in the AR gene by polymerase chain reaction (PCR) method. RESULTS: Ten alleles ranging from Z-10 (128 bp) to Z+8 (146 bp) in repeat number were identified. In cross-sectional studies, the prevalence of the Z+2 allele was higher than that of any other allele in patients with diabetic nephropathy (37.5% of patients in a microalbuminuria group, and 41.7% of those in a macroalbuminuria group including patients with chronic renal failure and maintenance haemodialysis treatment). Prevalence of the Z+2 allele was not increased in patients with diabetic retinopathy. In longitudinal Kaplan-Meier plots, the cumulative incidence of nephropathy was significantly associated with homozygosity for the Z+2 allele (log rank test, p = 0.031); respective prevalence of nephropathy after diabetes durations of 10 and 15 years was 42.9% and 100% in Z+2 homozygotes (n = 8), 17.6% and 27.4% in Z+2 heterozygotes (n = 44), and 6.1% and 17.4% in patients without the Z+2 allele (n = 49). However, occurrence of retinopathy was not influenced by the Z+2 allele (log rank test, p = 0.926). CONCLUSIONS: Homozygosity for the Z+2 allele was associated with accelerated early progression of diabetic nephropathy in Japanese type 1 diabetic patients. PMID- 12542726 TI - Clinical evaluation of pioglitazone in patients with type 2 diabetes using alpha glucosidase inhibitor and examination of its efficacy profile. AB - AIM: Pioglitazone is considered to reduce insulin resistance. This study was conducted to evaluate the efficacy, safety and clinical profile of pioglitazone in patients whose type 2 diabetes were poorly controlled with alpha-glucosidase inhibitor alone or alpha-glucosidase in combination with sulfonylurea. METHODS: Twenty patients with type 2 diabetes were treated with pioglitazone (30 mg q.d.) orally for 16 weeks. RESULTS: There were significant reductions in HbA1C, FPG and postprandial plasma glucose at week 16. As adverse events, oedema, hypoglycaemia like reaction, increases in LDH, CPK, etc. were noted. There was no significant change in TNF-alpha. Leptin levels increased significantly at week 16 and were still increasing 4 weeks after the treatment. Per cent body fat was almost constant throughout the study period. When efficacy was classified by demographic variables, pioglitazone was found to be more effective in the subjects who had a higher postprandial 2-h plasma glucose level, leptin level or per cent body fat value. CONCLUSION: Pioglitazone was considered to be effective when used in patients whose type 2 diabetes were poorly controlled with alpha-glucosidase inhibitor alone or alpha-glucosidase in combination with sulfonylurea. PMID- 12542728 TI - Pharmacological approaches to reducing allogeneic blood exposure. AB - This article discusses evidence for the role of pharmacological interventions such as the protease inhibitor aprotinin (Trasylol), lysine analogue anti fibrinolytics [tranexamic acid (Cyclokapron) and epsilon aminocaproic acid (Amicar)], DDAVP (Desmopressin) and recombinant Factor VIIa (NovoSeven), in preventing the need for blood and blood-component therapies after major (cardiac, hepatic and orthopaedic/trauma) surgery. The data show that aprotinin is consistently effective in reducing globally the transfusion burden in cardiac and hepatic surgical procedures. However, there are little data to support its use in routine elective orthopaedic surgery. Multiple studies have failed to show an increased risk for myocardial ischaemia or infarction with aprotinin, and there may even be a reduced incidence of perioperative stroke in patients undergoing cardiac surgery. An increased probability of a hypersensitivity reaction when the drug is readministered within a 6-month period remains a significant issue. The data for the lysine analogue anti-fibrinolytics show no evidence of efficacy in reducing the transfusion burden for epsilon aminocaproic acid and inconsistent results with tranexamic acid in cardiac and hepatic surgery. As with aprotinin therapy, there is a paucity of data to support their use in routine elective orthopaedic surgery. There are no data to support the routine use of DDAVP to reduce the transfusion burden. Limited data suggest that this drug may be effective when a defect in platelet function is demonstrated. This aspect deserves further investigation. Recombinant activated Factor VII (rFVIIa) has proven benefit for its licensed indication to reduce bleeding in haemophiliacs with inhibitors to Factors VIII and IX. Reports of benefit in other instances are largely anecdotal. Hence, at this time it is therefore speculative and premature to suggest whether there is a place for this agent in routine clinical practice. No adequately powered, placebo-controlled prospective studies are available to investigate the safety of the lysine analogues, DDAVP or rFVIIa in cardiac, hepatic or orthopaedic surgery. PMID- 12542729 TI - Nucleic acid technology screening of Australian blood donors for hepatitis C and human immunodeficiency virus-1 RNA: comparison of two high-throughput testing strategies. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to compare the performance of two high-throughput strategies for hepatitis C virus (HCV) and human immunodeficiency virus-1 (HIV-1) RNA nucleic acid technology (NAT) screening in a volunteer blood-donor population. MATERIALS AND METHODS: The semiautomated Chiron Procleix HIV-1/HCV transcription mediated amplification (TMA) assay was used to screen 1 439 765 donations in two different testing configurations. Three sites (termed PDT sites) performed a mixture of individual donation (ID) and minipool (MP) testing, where 1 113 288 donations were screened as pools of 24 and an additional 32 003 donations were screened in ID format. A further two sites (termed SDT sites) screened 294 474 donations exclusively in ID format. RESULTS: A significantly higher proportion of initial NAT reactives that failed to react on follow-up testing [termed non-repeatably reactive (NRR)] was observed for ID testing at SDT sites than at PDT sites (0.082 vs. 0.047%: P < 0.01). Within the PDT sites, however, there was no significant difference between the NRR rate for MP or ID samples (0.037 vs. 0.047%; not significant). There was a significant difference in failed run rates between PDT and SDT sites (P < 0.01), with PDT sites having a higher run failure rate owing to non-amplification of the internal control. The PDT sites also had a significantly higher overall invalid sample rate. However, the invalid sample rate, specifically caused by known equipment failure, was significantly higher in the SDT sites, possibly attributable to greater usage (P < 0.0001). Four HCV NAT-positive/antibody-negative samples were identified in the course of the study. CONCLUSIONS: The comparison of the performance of PDT with SDT sites identified only minor differences that did not adversely impact on the timely release of blood products. Although both ID and MP strategies showed excellent specificity, irrespective of site configuration, the significantly increased NRR rate, observed exclusively for ID testing performed at SDT sites, indicates a potential for contamination that may limit the number of samples that can optimally be processed using ID testing. The performance data for ID testing in particular should serve as a useful benchmark for evaluating candidate NAT systems that are fully automated. PMID- 12542730 TI - Collaborative study to calibrate hepatitis C virus genotypes 2-6 against the HCV International Standard, 96/790 (genotype 1). AB - BACKGROUND AND OBJECTIVES: A major requirement of a hepatitis C virus (HCV) RNA nucleic acid amplification technology (NAT) assay validation is the ability of the assay to detect the six major genotypes of HCV with equivalent sensitivities. The aim of this study was to characterize and calibrate an HCV genotype panel for use in such studies. MATERIALS AND METHODS: Panels consisting of the first International Standard (IS) for HCV RNA NAT assays (96/790; HCV genotype 1a) and isolates of genotypes 2-6 were sent to 17 laboratories worldwide which use a variety of NAT tests, both qualitative and quantitative. The HCV RNA content of each panel member was determined and the mean titre calculated in International Units/ml (IU/ml). RESULTS: The calculated mean titres (calibrated against the HCV International Standard), in log10 IU/ml, of the genotype 2-6 samples were 3.99, 3.81, 4.14, 4.18 and 4.61, respectively. CONCLUSIONS: An HCV genotype panel, calibrated in IU/ml, has been established and should be valuable for assay validation. All the genotypes were detected by all the assays used, but it was not possible to demonstrate that the genotypes were detected with equal efficiencies. PMID- 12542731 TI - The influence of platelet additive solutions on cytokine levels and complement activation in platelet concentrates during storage. AB - BACKGROUND AND OBJECTIVES: The accumulation of platelet-derived cytokines in platelet concentrates (PC) during storage may contribute towards non-haemolytic transfusion reactions (NHTR). We investigated the effect of platelet storage medium on platelet activation, complement activation and cytokine levels in leucocyte-reduced PC. MATERIALS AND METHODS: Hyperconcentrated platelets (3000 6000 x 109/l) were collected by apheresis and diluted in 100% plasma, 70% PASIII, or 70% or 80% PASIII supplemented with magnesium and potassium (PAS IIIM). RESULTS: Levels of transforming growth factor-beta (TGF-beta) and regulated on activation, normal, T-cell expressed, and secreted (RANTES) increased during storage, as did the expression of P-selectin (CD62P), and were highest in PC stored in PASIII. In PC stored in PASIIIM, however, levels of TGF-beta and RANTES were not significantly different from PC stored in plasma. Levels of CD62P expression, however, remained higher in PASIIIM PC than in those stored in plasma by day 5, but were lower than PC stored in PASIII. C3a des arg levels increased during storage in all media with the exception of PASIII and, on day 7, were higher in PC stored in plasma compared to PC stored in the other media. CONCLUSIONS: Our results indicate that replacing plasma in PC with unmodified PASIII for storage results in higher levels of platelet-derived cytokines in PC. Furthermore, it appears that the nature of the medium used for storage of PC has a significant impact on platelet activation and cytokine levels of the PC. These implications should be taken into account when considering replacement of plasma with PAS. PMID- 12542732 TI - Inactivation of viral and prion pathogens by gamma-irradiation under conditions that maintain the integrity of human albumin. AB - BACKGROUND AND OBJECTIVES: The administration of therapeutic plasma protein concentrates has been associated with the real risk of transmitting viral diseases and the theoretical risks of prion transmission. Our objective was to determine if gamma-irradiation can inactivate viral or prion infectivity without damaging a protein biotherapeutically. MATERIALS AND METHODS: Human albumin 25% solution, spiked with four model viruses (including porcine parvovirus) or with brain homogenate from scrapie-infected hamsters, was gamma-irradiated at constant low-dose rates and assayed for viral and prion infectivity or for albumin integrity. RESULTS: At a radiation dose of 50 kGy, viruses were inactivated by >/= 3.2 to >/= 6.4 log10 and scrapie by an estimated 1.5 log10, whereas albumin was only moderately aggregated and fragmented. CONCLUSIONS: gamma-Irradiation can preferentially inactivate viral and prion pathogens without excessive damage to albumin structure. PMID- 12542733 TI - Efficacy, tolerability, safety and pharmacokinetics of a nanofiltered intravenous immunoglobulin: studies in patients with immune thrombocytopenic purpura and primary immunodeficiencies. AB - BACKGROUND AND OBJECTIVES: A nanofiltration step with the capacity to reduce blood-borne pathogens was introduced into the manufacturing process of intravenous immunoglobulin (IVIG). In order to demonstrate the efficacy, safety and pharmacokinetics of the modified product, we conducted Phase II/III studies comparing the nanofiltered IVIG (IVIG-N) with its parent product, Sandoglobulin, in patients with chronic immune thrombocytopenic purpura (ITP) and primary immunodeficiencies (PID). MATERIALS AND METHODS: Patients with ITP (n = 27) with platelet counts of < 20 x 10(9)/l were treated with Sandoglobulin or IVIG-N infusions at a dose of 0.4 g/kg body weight on five consecutive days. The primary efficacy end-point was the number of patients with an increase in platelet counts to > 50 x 10(9)/l. Secondary end-points were time to and duration of response, and regression of bleeding. Patients with PID (n = 36) were treated for 6 months with Sandoglobulin or IVIG-N at doses of 0.2-0.8 g/kg, infused at 3- or 4-week intervals. The primary end-point was the number of days absent from school/work. Secondary end-points were hospitalization, use of antibiotics and feeling of well being. In both studies, tolerability was assessed by recording of adverse events and laboratory determinations. Viral safety was ascertained by serology supplemented with nucleic acid detection methods. Pharmacokinetics were analysed in patients with PID using serum concentration-time data for immunoglobulin G (IgG), and IgG antibodies to hepatitis B surface antigen (anti-HBsAg). RESULTS: In the ITP study, the primary end-point was met by 12/16 patients on IVIG-N and by 10/10 patients on Sandoglobulin (P = 0.123). A shift towards lesser bleeding intensity was seen in both groups. In the PID study, seven of 18 patients on IVIG N and six of 16 patients on Sandoglobulin missed days at work/school, with monthly mean absences of 0.4 and 0.5 days (P = 0.805). The feeling of well-being was comparable in both groups. In the ITP study, adverse events with a causal relationship to medication were suspected in six patients on IVIG-N and in seven on Sandoglobulin. In the PID study, three patients on IVIG-N and two on Sandoglobulin experienced possible drug-related adverse events. In both studies, serological and polymerase chain reaction (PCR) tests gave evidence for virus safety. Pharmacokinetics showed constant peak and trough serum IgG levels in all patients, indicating almost steady-state conditions for both formulations. The overall half-life (t1/2) for total IgG was 33 +/- 17 days in the IVIG-N arm and 25 +/- 16 days in the Sandoglobulin arm; for anti-HBsAg t1/2, values were 17 +/- 7 and 17 +/- 9 days, respectively. CONCLUSIONS: IVIG-N is efficacious, well tolerated and safe in patients with ITP and PID. Its pharmacokinetic properties were comparable to those of Sandoglobulin. PMID- 12542734 TI - Large-scale production and properties of human plasma-derived activated Factor VII concentrate. AB - BACKGROUND AND OBJECTIVES: An activated Factor VII (FVIIa) concentrate, prepared from human plasma on a large scale, has to date not been available for clinical use for haemophiliacs with antibodies against FVIII and FIX. In the present study, we attempted to establish a large-scale manufacturing process to obtain plasma-derived FVIIa concentrate with high recovery and safety, and to characterize its biochemical and biological properties. MATERIALS AND METHODS: FVII was purified from human cryoprecipitate-poor plasma, by a combination of anion exchange and immunoaffinity chromatography, using Ca2+-dependent anti-FVII monoclonal antibody. To activate FVII, a FVII preparation that was nanofiltered using a Bemberg Microporous Membrane-15 nm was partially converted to FVIIa by autoactivation on an anion-exchange resin. The residual FVII in the FVII and FVIIa mixture was completely activated by further incubating the mixture in the presence of Ca2+ for 18 h at 10 degrees C, without any additional activators. For preparation of the FVIIa concentrate, after dialysis of FVIIa against 20 mm citrate, pH 6.9, containing 13 mm glycine and 240 mm NaCl, the FVIIa preparation was supplemented with 2.5% human albumin (which was first pasteurized at 60 degrees C for 10 h) and lyophilized in vials. To inactivate viruses contaminating the FVIIa concentrate, the lyophilized product was further heated at 65 degrees C for 96 h in a water bath. RESULTS: Total recovery of FVII from 15 000 l of plasma was approximately 40%, and the FVII preparation was fully converted to FVIIa with trace amounts of degraded products (FVIIabeta and FVIIagamma). The specific activity of the FVIIa was approximately 40 U/ micro g. Furthermore, virus-spiking tests demonstrated that immunoaffinity chromatography, nanofiltration and dry heating effectively removed and inactivated the spiked viruses in the FVIIa. These results indicated that the FVIIa concentrate had both high specific activity and safety. CONCLUSIONS: We established a large-scale manufacturing process of human plasma-derived FVIIa concentrate with a high yield, making it possible to provide sufficient FVIIa concentrate for use in haemophiliacs with inhibitory antibodies. PMID- 12542735 TI - Cost-effectiveness of leucocyte depletion of red-cell transfusions for patients undergoing cardiac surgery. PMID- 12542737 TI - Detection of platelet-reactive antibodies in patients who are refractory to platelet transfusions, and the selection of compatible donors. PMID- 12542736 TI - Lymphocyte T subsets and natural killer cells in Italian and Philippino blood donors. AB - BACKGROUND AND OBJECTIVES: The characterization of lymphocyte subsets in blood donors has been utilized to determine the normal ranges that can be related to race. A study was performed in blood donors from two racial groups - Caucasian (Italians) and Asian (Philippinos) - to define respective T-lymphocyte subsets and levels of cytokines. MATERIALS AND METHODS: Ninety-two blood donors (46 Italians and 46 Philippinos) were enrolled. Blood count and immunophenotyping of lymphocytes by flow cytometry were carried out, and cytokine production was tested in six blood donors of each group. RESULTS: Philippino blood donors showed a significantly higher mean value of leucocytes (P = 0.01) and lymphocytes (P < 0.001) than Italians. The mean absolute count of lymphocyte subsets CD3- CD16+ CD56+ and CD3+ CD8+ were both significantly higher in Philippino than in Italian subjects, respectively, P < 0.01 and P < 0.0001. Philippinos showed a statistically significant higher frequency of lymphocytes producing interferon gamma (IFN-gamma) compared to Italians (P = 0.02). CONCLUSIONS: T-lymphocyte subsets in Italian and Philippino blood donors seem to be correlated to ethnic background. The higher levels of CD3+ CD8+ T cells, natural killer (NK) cells and IFN-gamma-producing cells found in Philippinos suggest leucoreduction in Asian blood donors. PMID- 12542739 TI - Genetic epidemiology of rheumatoid arthritis. AB - Building on the spectacular success of molecular genetics in defining the biological basis of many rare single gene disorders over the past decade, epidemiologists have turned their attention to unravelling the complex genetic mysteries of common disorders, such as rheumatoid arthritis (RA). As a prelude to any such endeavour it is obviously important to establish that there is a significant genetic component to the disease. The classical approaches of twin and other family recurrence risk studies, coupled with prevalence studies in different ethnic and migrant populations, have been used to estimate the environmental and genetic contributions to RA. However, developing a consensus on these estimates has proved difficult, thereby providing an early warning to the unwary investigator that the road to gene discovery in RA is likely to be a rough ride. PMID- 12542740 TI - Common HLA-B8-DR3 haplotype in Northern India is different from that found in Europe. AB - The aim of this study was to determine whether a common diabetic haplotype, including human leukocyte antigen (HLA)-B8 and HLA-DR3, in Northern India is the same haplotype as the European HLA-B8-DR3 haplotype. DNA samples from Northern Indian subjects selected on the basis of HLA-B8 and HLA-DR3 were tested for microsatellite and single nucleotide polymorphism alleles throughout the major histocompatibility complex (MHC). It was found that the Indian samples represent a conserved haplotype in which all alleles were shared by Indian subjects with HLA-B8 and HLA-DR3, but were different to those that are characteristic of the European 8.1 ancestral haplotype. The Indian and European haplotypes share HLA B*0801, HLA-DRB1*0301 and HLA-DQB1*02 but differ for subtypes of HLA-Cw*07 and HLA-DRB3 and all central MHC alleles tested. In contrast, Indian subjects selected on the basis of HLA-B58 ( 1-17) and HLA-DR3 shared the same alleles at other MHC loci as have been described in the common Chinese haplotype with HLA B58/17 and HLA-DR3. A third haplotype, HLA-B50/21 and HLA-DR3, was also found to be highly conserved but shares little in common with the other two HLA-DR3 containing Indian haplotypes. PMID- 12542741 TI - Comprehensive hereditary hemochromatosis genotyping. AB - Hereditary hemochromatosis (HH) is an iron-overload disease common in populations of Northern European origin. Patients display increased iron absorption leading to excessive iron deposition and potential multiorgan failure. Using polymerase chain reaction sequence-specific primer (PCR-SSP) technology, we have developed an HH diagnosis assay capable of detecting 19 non-synonymous HFE mutations (including a previously unreported mutation, V295A) and several TFR2, SLC11A3 and H ferritin alleles implicated in HH. As part of the validation process, 159 UK renal donors were genotyped to determine HH allele frequencies in the UK population. The alleles nominally identified as HFE*01 (C282Y), HFE*02 (H63D) and HFE*03 (S65C) were found at frequencies of 0.085, 0.173 and 0.009, respectively. All other potential HH-associated alleles were absent, confirming their low prevalence in this population. This assay enables comprehensive routine HH genotyping, producing rapid, accurate and reproducible results at low cost. PMID- 12542742 TI - Role of the autoimmune regulator (AIRE) gene in alopecia areata: strong association of a potentially functional AIRE polymorphism with alopecia universalis. AB - Alopecia areata is characterized by a reversible form of patchy or complete hair loss associated with T-cell infiltration of hair follicles. The lifetime disease risk of approximately 1.4% in the general population is increased to more than 30% in autoimmune polyendocrinopathy candidiasis ectodermal dysplasia syndrome (APECED), a recessive condition resulting from a mutation of the autoimmune regulator (AIRE) gene on chromosome 21q22.3. Aire protein is thought to have transcriptional regulatory activity but its role is not well defined at present. In this study, we have examined the possible involvement of AIRE in the pathogenesis of alopecia areata. On screening the AIRE coding sequence, we identified 20 variants. Two of these at positions, G961C and T1029C, give rise to amino acid changes, S278R and V301A, located in the DNA-binding segment (SAND) and PHD1 zinc finger motif, respectively. We found no difference in the frequency of the AIRE T1029C polymorphism between the control and patient groups. We genotyped 202 alopecia areata patients and 175 matched Caucasian controls for the AIRE G961C alleles. The frequency of the rare allele (961G) was 0.08 in the controls and there was a significant increase to 0.13 in alopecia areata overall and 0.20 in severe disease (alopecia universalis). We found no association between the AIRE G961G variant and mild (patchy) alopecia areata or alopecia totalis. However, the AIRE 961G allele is a potent risk factor (> 3) for the severest form of alopecia areata, and for disease of early age at onset (at 30 years). The change from serine to arginine in the SAND domain of AIRE protein may have a significant effect on AIRE DNA-binding activity. Moreover, our results could provide a rational explanation of the unusually high frequency of AA in APECED patients, supporting the concept of AA as an autoimmune disease. PMID- 12542743 TI - HLA polymorphism in Bulgarians defined by high-resolution typing methods in comparison with other populations. AB - In the present study we analyzed for the first time HLA class I and class II polymorphisms defined by high-resolution typing methods in the Bulgarian population. Comparisons with other populations of common historical background were performed. Most HLA-A, -B, -DRB alleles and haplotypes observed in the Bulgarian population are also common in Europe. Alleles and haplotypes considered as Mediterranean are relatively frequent in the Bulgarian population. Observation of Oriental alleles confirms the contribution of Asians to the genetic diversity of Bulgarians. The use of high-resolution typing methods allowed to identify allele variants rare for Europeans that were correlated to specific population groups. Phylogenetic and correspondence analyses showed that Bulgarians are more closely related to Macedonians, Greeks, and Romanians than to other European populations and Middle Eastern people living near the Mediterranean. The HLA-A, B,-DRB1 allele and haplotype diversity defined by high-resolution DNA methods confirm that the Bulgarian population is characterized by features of southern European anthropological type with some influence of additional ethnic groups. Implementation of high-resolution typing methods allows a significantly wider spectrum of HLA variation to be detected, including rare alleles and haplotypes, and further clarifies the origin of Bulgarians. PMID- 12542744 TI - Evidence for discoordinate regulation of the HLA-DPB1 gene. AB - Characterization of cell lines derived from patients with type II bare lymphocyte syndrome, a pathological state in which the constitutive and inducible expression of HLA class II antigens is lacking, has permitted the identification of several trans-acting factors involved in the coordinated regulation of HLA class II genes. Although an increasing body of evidence has pointed to the existence of a discoordinate regulation of HLA class II loci, the mechanisms underlying such regulation are essentially unknown. In the present study, 45.EM2, a mutant lymphoblastoid cell line with a new pattern of HLA discoordinate expression is characterized. 45.EM2 expresses HLA-DR and -DQ but fails to express HLA-DP. The absence of HLA-DP expression in 45.EM2 is the result of a transcriptional defect, leading to a lack of DPB1 mRNA. By contrast, DPA1 transcription in this LCL is not impaired. The characteristics of 45.EM2 described here suggest the existence of a specific trans-acting factor involved in the control of DPB1 gene expression. PMID- 12542745 TI - Enhanced binding of low-affinity antibodies interacting simultaneously with targeted cell surface molecules and Fc receptor. AB - The possibility that different forms of class I molecules might be expressed on the cell surface of lymphocytes has been investigated periodically over the past several decades. A series of major histocompatibility complex (MHC) class I specific monoclonal antibodies, including the commonly used antibodies 64-3-7 and 25-D1.16, bind B cells differentially, suggesting the existence of differentially expressed class I-associated cell surface determinants on B lymphocytes. However, the ability of antibodies to bind cells is determined by the sum of interactions between the antibodies and the molecules expressed on the cell surface. The interactions of class I-specific antibodies with B cells were dissected, revealing dual specificity of the antibodies for the targeted class I molecules, as well as to Fc receptors preferentially expressed by B cells. We demonstrate that antibodies simultaneously bind targeted class I molecules and Fc receptors expressed on the surface of B cells. Simultaneous binding to two cell surface structures significantly enhances the class I-specific binding pattern of certain antibodies by increasing their avidity, leading to apparent cell-specific differences in MHC expression patterns. We conclude that no differences in MHC structures need be postulated to account for the observed binding patterns. PMID- 12542746 TI - A new allele within the transmembrane region of the human MICA gene with seven GCT repeats. AB - Major histocompatibility complex class I chain-related genes (MIC) belong to a multicopy gene family located within the HLA class I region of chromosome 6. They encode for proteins that have a completely different organization, expression, and products from classical HLA class I gene products. One member of this family is the MICA gene, which is characterized by its high degree of polymorphism, with over 50 MICA alleles described. Moreover, MICA exon 5 presents a microsatellite polymorphism consisting of a variable number of GCT repeats that encode for 4, 5, 6, 9, or 10 alanine residues, and a variant (MICA A5.1) that includes a nucleotide insertion (GCT-->GGCT). In this study, we report a novel allele in the transmembrane region of the MICA gene consisting of seven GCT repeats found in a family based study of MICA polymorphism in celiac disease. PMID- 12542747 TI - Identification of HLA-A*0249 and confirmation of A*2615. AB - A new HLA-A*02 allele, A*0249, and the confirmatory sequence of A*2615 are here reported. Both alleles were detected by irregular patterns during routine molecular typing. A*0249 showed two mismatches with A*02011 in exon 3 at positions 538 and 539, changing amino acid 156 from leucine to arginine. Remarkable was a mismatch in exon 4, at position 779, where the C of A*0201 was changed to A in A*0249. As A*0209 has the same substitution, A*0249 may have arisen from A*0209. The A*2615 allele was identical to A*2601 in exons 2 and 3 except for a single nucleotide difference at nucleotide position 180, changing codon 36 from phenylalanine to leucine. This is a unique amino acid change, as in all the class I alleles a phenylalanine was identified at this position. PMID- 12542748 TI - Major histocompatibility complex class II DOA sequences from three Antarctic seal species verify stabilizing selection on the DO locus. AB - To provide additional support for the sequence conservation and hence the regulatory role of the MHC class II DOA locus, we obtained the nucleotide sequences of exon 2 and exon 3, along with the intervening intron, of the Ross seal, and sequences from the exon 2 region from the Weddell and leopard seals. These are the first reports of the sequences of this locus from a carnivore species. The results demonstrate strong conservation among mammals for the exon sequence and produce a gene genealogy that is consistent in topology with a species tree. PMID- 12542749 TI - New HLA-DRB1*07 allele identified from microlymphocytotoxicity/SSP discrepancies. AB - A HLA-DRB1*07 variant allele has been identified in a cadaver kidney donor. Serological typing using monoclonal antibodies detected HLA-DR4 and HLA-DR7. HLA class II DNA typing using sequence-specific primer (PCR-SSP) polymerase chain reaction only detected DRB1*04, while sequence-specific oligonucleotide (PCR-SSO) polymerase chain reaction confirmed the presence of both DRB1*04 and DRB1*07 alleles, although two extra reactions were also found. Exon 2 of the HLA-DRB1*07 was isolated using allele-specific PCR, then cloned and sequenced. Four mutations, at positions 170 (T --> C), 171 (C --> T), 174 (C --> G), and 179 (C - > A), were observed. These mutations changed codons 57 and 60 (V --> A; S --> Y, respectively). This amino acid sequence at position 56-61 is only found in DRB1*0811. PMID- 12542750 TI - Nomenclature for factors of the HLA system, update July 2002. PMID- 12542751 TI - Nomenclature for factors of the HLA system, update August 2002. PMID- 12542752 TI - Nomenclature for factors of the HLA system, update September 2002. PMID- 12542753 TI - Tissue Typing and Immunosuppression Symposium. Ankara, Turkey, 19-21 June 2002. Abstracts. PMID- 12542792 TI - Anti-inflammatory activities of human lactoferrin in acute dextran sulphate induced colitis in mice. AB - In this study, we investigated the anti-inflammatory effects of orally administered human lactoferrin (hLF) and two peptides, based on the bactericidal region of hLF (HLD1 and HLD2), on the course of experimental colitis. Acute colitis was induced in C57Bl/6 mice by giving 5% dextran sulphate (DX) in the drinking water. The mice were killed after 2 or 7 days of DX exposure. The animals were given hLF or the peptides orally twice a day (2 mg/dose/mouse) during the DX exposure. In the control animals, the hLF or the peptides were replaced by bovine serum albumin or water. The appearance of occult blood in the faeces and macroscopic rectal bleeding were significantly delayed and partly reduced in the hLF-treated animals compared with the control animals. The shortening of the colon, a pathological effect of DX exposure, was significantly less pronounced in the hLF-treated group compared with the control group. Also, the interleukin-1beta (IL-1beta) levels in the blood were significantly diminished in this group after 2 days of DX exposure. A significantly lower crypt score was observed in the distal part of the colon in the hLF-treated group compared with the control group. Also, significantly reduced numbers of CD4 cells, F4/80-positive macrophages and tumour necrosis factor-alpha-producing cells were detected by immunohistochemistry in the distal colon of the hLF treated animals compared with the control animals after 7 days of DX exposure. A reduction was also observed concerning the IL-10-producing cells in the middle colonic submucosa. The HLD1 and HLD2 treatment, which was carried out for 2 days, only gave results almost identical to those of hLF, concerning clinical parameters after the 2 days of DX exposure. An even stronger effect was observed for HLD2, regarding decreased occult blood in the faeces and colon length. Our results show that perorally given hLF mediates anti-inflammatory effects on the DX-induced acute colitis, and further suggest that the bactericidal region of the hLF molecule may be involved in these activities. PMID- 12542793 TI - Mesenchymal stem cells inhibit and stimulate mixed lymphocyte cultures and mitogenic responses independently of the major histocompatibility complex. AB - We aimed to study the effects of mesenchymal stem cells (MSCs) on alloreactivity and effects of T-cell activation on human peripheral blood lymphocytes (PBLs) in vitro. MSCs were expanded from the bone marrow of healthy subjects. MSCs isolated from second to third passage were positive for CD166, CD105, CD44, CD29, SH-3 and SH-4, but negative for CD34 and CD45. MSCs cultured in osteogenic, adipogenic or chondrogenic media differentiated, respectively, into osteocytes, adipocytes or chondrocytes. MSC added to PBL cultures had various effects, ranging from slight inhibition to stimulation of DNA synthesis. The stimulation index (SI = (PBL + MSC)/PBL) varied between 0.2 and 7.3. The SI was not affected by the MSC dose or by the addition of allogeneic or autologous MSCs to the lymphocytes. Suppression of proliferative activity was observed in all experiments after the addition of 10,000-40,000 MSCs to mixed lymphocyte cultures (MLCs). Lymphocyte proliferation was 10-90%, compared with a control MLC run in parallel without MSCs. In contrast, the addition of fewer MSCs (10-1000 cells) led to a less consistent suppression or a marked lymphocyte proliferation in several experiments, ranging from 40 to 190% of the maximal lymphocyte proliferation in control MLCs. The ability to inhibit or stimulate T-cell alloresponses appeared to be independent of the major histocompatibility complex, as results were similar using 'third party' MSCs or MSCs that were autologous to the responder or stimulating PBLs. The strongest inhibitory effect was seen if MSCs were added at the beginning of the 6 day culture, and the effect declined if MSCs were added on day 3 or 5. Marked inhibitory effects of allogeneic and autologous MSCs (15,000) were also noted after mitogenic lymphocyte stimulation by phytohaemagglutinin (median lymphocyte proliferation of 30% of controls), Concanavalin A (56%) and protein A (65%). Little, if any, inhibition occurred after stimulation with pokeweed mitogen. Low numbers of MSCs (150 cells) were unable to inhibit mitogen-induced T cell responses. MSCs have significant immune modulatory effects on MLCs and after mitogenic stimulation of PBL. High numbers of MSCs suppress alloreactive T cells, whereas very low numbers clearly stimulated lymphocyte proliferation in some experiments. The effect of a larger number of MSCs on MLCs seems more dependent on cell dose than histocompatibility and could result from an 'overload' of a stimulatory mechanism. PMID- 12542794 TI - Self-peptides with intermediate capacity to bind and stabilize MHC class I molecules may be immunogenic. AB - Thirty self-peptides were selected on the basis of their predicted binding to H 2b molecules. The binding of peptides was ascertained experimentally by biochemical (KD measurements) and cellular [major histocompatibility complex class I (MHC-I) stabilization] assays. A weak, but significant, correlation between KD measurements and MHC-I stabilization was observed. Mice (n = 99) were immunized with individual peptides. Twenty-eight peptides were found to induce peptide-specific cytotoxic activity, and a total of 84 mice developed significant cytotoxic T lymphocyte (CTL) responses after immunization. Only one of the 21 mice immunized with high-affinity peptides developed a peptide-specific CTL response of 29 lytic units per 106 splenocytes, whereas 11 of the 42 mice immunized with intermediate-affinity peptides developed peptide-specific CTL responses at this level (P < 0.05). These observations suggest the absence of tolerance towards most MHC-I-restricted self-peptides and that strong antiself immunity can be generated preferentially towards self-peptides with an intermediate affinity for MHC-I. These data should be considered in the design of tumour vaccines based on MHC-I-binding self-peptides. PMID- 12542795 TI - Downregulation of mafB expression in T-helper cells during early differentiation in vitro. AB - We have studied the expression of a human homologue of mafB (maf-1), a member of the family of large maf transcription factors. In support of the suggested key role that mafB expression plays in differentiating macrophages, we found mafB to be expressed at a very high level in monocytic U937 and THP-1 cell lines. However, we show here that mafB transcription is not restricted to myeloid cells but can also be detected in lymphoid cells, indicating transcriptional plasticity during haematopoiesis. In conclusion, strong proliferative signals mediated by T cell activation and interleukins (IL-4 and IL-12) downregulate the mafB messenger RNA transcript level when resting naive CD4+ T-helper cells enter the differentiation pathway in vitro. PMID- 12542796 TI - Downregulation of mac-1 expression in monocytes by surface-bound IgG. AB - Physical and functional association between the beta2-integrin Mac-1 (CD11b/CD18) and receptors of immunoglobulin G (IgG) (FcgammaRs) has been previously reported. In this study, we examined the modulation of Mac-1 expression by IgG in different leucocyte populations. Our data show that human monocytes, but not neutrophils, macrophages, dendritic or natural killer cells, downregulate the expression of Mac-1 after overnight exposure to surface-bound IgG. This effect, which requires at least 6 h of incubation, is not associated with a general downmodulation of membrane antigens, and is selectively induced by immobilized IgG (iIgG), as the stimulation of monocytes with N-formyl-methionyl-leucyl-phenylalanine, lipopolysaccharide, tumour necrosis factor-alpha (TNF-alpha) or soluble IgG did not modify the Mac-1 expression after 18 h in culture. The loss of Mac-1 was completely prevented by blocking antibodies (Abs) directed to FcgammaRII (CD32) or CD18. On the other hand, the serine protease inhibitor, phenyl methyl sulphonyl fluoride, but not inhibitors of cysteine proteases or neutral endopeptidases, partially prevented the downregulation of Mac-1 by iIgG. Monocytes cultured overnight on iIgG exhibited a dramatic decrease in their capacity to ingest zymosan particles that could be attributed to the reduced expression of Mac-1. However, there was no inhibition of TNF-alpha production induced by zymosan, suggesting that Mac-1-dependent responses require different levels of its expression to be fully activated. PMID- 12542797 TI - Intranasal Cry1Ac protoxin is an effective mucosal and systemic carrier and adjuvant of Streptococcus pneumoniae polysaccharides in mice. AB - Streptococcus pneumoniae is a major respiratory pathogen in infants, children and the elderly. Available parenteral anti-pneumococcal vaccines based on type specific capsular polysaccharides (CPSs) are useful in adults but do not elicit protective immunity in infants and young children. To enhance their immunogenicity, pneumococcal CPSs conjugated to proteins are being developed. Mucosal vaccines may induce mucosal and systemic immune responses, but their development has been hampered by the lack of effective, inexpensive innocuous mucosal adjuvants or immunogenic vaccine carriers. We have demonstrated that the recombinant Cry1Ac protoxin from Bacillus thuringiensis is highly immunogenic and has mucosal and systemic adjuvant effects on proteins coadministered in mice. In this work, we evaluated Cry1Ac as a carrier and adjuvant of S. pneumoniae CPS for the induction of mucosal and systemic antibody responses after intranasal and intraperitoneal immunization in mice. Our results demonstrate that intranasal application of pneumococcal polysaccharides either coadministered or conjugated with Cry1Ac induces higher systemic and mucosal specific antibody responses than those elicited by pneumococcal polysaccharides alone. Adjuvant effects of Cry1Ac on polysaccharides may be appropriate for vaccine design. PMID- 12542798 TI - Single-cell analysis of Ets-1 transcription factor expression during lymphocyte activation and apoptosis. AB - The Ets-1 proto-oncogene is a prototype member of Ets family of transcription factors. It is preferentially expressed in lymphoid cells, where it is essential for the maintenance of the normal pool of resting T and B cells. We have investigated the protein expression of the Ets-1 transcription factor during the activation and apoptosis of T and B cells by flow cytometry and confocal microscopy. Cells of the thymus, spleen and bursa expressed high levels of Ets-1 protein, while resting peripheral blood mononuclear cells had lower Ets-1 expression. Activation and proliferation of T cells induced the upregulation of Ets-1. alphabeta-T cells were found to upregulate Ets-1 expression more than gammadelta-T cells. Increased Ets-1 protein expression was located predominantly in the perinuclear area. In contrast, during apoptosis, Ets-1 expression was downregulated. Collectively, our results indicate that Ets-1 expression can be accurately determined by flow cytometry and confocal microscopy. Ets-1 expression level and distribution are differentially controlled in resting, activated and apoptotic lymphocytes. PMID- 12542799 TI - Staphylococcal enterotoxin B upregulates fas-mediated apoptosis of peripheral blood mononuclear cells in childhood atopic dermatitis. AB - Staphylococcal infection-producing superantigens, such as staphylococcal enterotoxin B (SEB), are presumed to play an important role of inflammatory processes in atopic dermatitis (AD). The aim of this study was to elucidate the apoptotic response of peripheral blood mononuclear cells (PBMCs) from children with AD. PBMCs from AD children were sampled and cultured with SEB stimulation. Levels of apoptosis and Fas expression were measured using flow cytometry; the soluble Fas ligand (sFasL) was also measured using the enzyme-linked immunosorbent assay method, and the expression of FasL in PBMCs was observed using reverse transcriptase-polymerase chain reaction. There was no difference in the initial levels of apoptosis and Fas expression in precultured PBMCs of AD patients and healthy donors. After culturing for 48 h under SEB stimulation, the apoptosis level and Fas expression were significantly upregulated in the PBMCs from AD children compared with that from the normal controls. In patients, the sFasL was significantly increased, and the expression of FasL was observed in messenger RNA of peripheral monocytes. These results suggest that the Fas/FasL system is involved in the apoptosis induced by SEB in AD, with simultaneous increases in sFasL and expression of FasL. PMID- 12542800 TI - Effects of anti-neutrophil cytoplasm autoantibodies on tissue factor activity by HL-60 cells in vitro. AB - The effects of cytoplasmic anti-neutrophil cytoplasm autoantibodies (C-ANCA) and perinuclear ANCA (P-ANCA) immunoglobulin G (IgG) on tissue factor (TF) activity using HL-60 cells in vitro were compared with those of medium, lipopolysaccharide (LPS) and control IgG. Cells were also incubated with both ANCA IgG and control IgG in the presence of a submaximal concentration of LPS capable of upregulating TF procoagulant activity (TF-PCA) measured in arbitrary units of TF equivalent (AU-TFEq). The purpose was to search for an additive effect between LPS and ANCA IgG. All IgG preparations increased HL-60 cell TF-PCA in comparison with the medium. When cells were incubated with P-ANCA IgG and LPS (1 micro g/ml), a larger increase was seen (151.23 +/- 31.6 SEM (standard error of the mean) AU TFEq) than when incubated with control IgG plus LPS (91.01 +/- 18.4 SEM AU-TFEq; P < 0.005), P-ANCA IgG alone (73.68 +/- 12.7 SEM AU-TFEq; P < 0.005) or LPS (1 micro g/ml) (58.11 +/- 7.9 SEM AU-TFEq; P < 0.005). There was concordance between PCA and TF total antigen content by enzyme-linked immunosorbent assay (ELISA). The fact that P-ANCA IgGs upregulate the function of TF in HL-60 cells in combination with LPS adds to information regarding the possible role of ANCAs in the enhancement of TF by different cells, although it does not support the fact that ANCAs alone play a role in mononuclear cell TF upregulation. The additive effects of LPS underline the possible role of pro-inflammatory stimuli in the pathogenesis of ANCA-associated diseases. PMID- 12542801 TI - Novel specificity of anti-U1A autoimmune patient sera. AB - We have previously described a novel complex of proteins which contains the U1snRNP-A protein (U1A) but no other small nuclear ribonucleoprotein particle (snRNP) components (O'Connor et al., RNA 1997;3:1444-55). Antibodies to this novel complex inhibit both splicing and polyadenylation in vitro of a test pre mRNA (O'Connor et al., RNA 1997;3:1444-55; Lutz et al., RNA 1998;4:1493-9). This novel complex of proteins was identified using an unusual mouse monoclonal antibody (MoAb), called MAb 12E12. We were interested to know if autoimmune patient sera were similar to this MoAb. We have discovered a novel specificity of systemic lupus erythematosus patient sera reminiscent of MAb 12E12 in that the patient serum, like 12E12, (1) does not recognize U1A when bound to U1 RNA, (2) recognizes primarily the epitopes in the amino-terminal third of the protein, including RNA recognition motif 1 (RRM1) and (3) inhibits in vitro polyadenylation. These findings may lead to the discovery of previously undescribed autoantigens as components of the novel protein complex, and may provide insight into autoimmune diseases. PMID- 12542802 TI - Clonal analysis of a human antimouse antibody (HAMA) response. AB - Circulating human antimouse antibodies (HAMAs) directed to mouse immunoglobulin G (IgG) are clinically significant, compromising mouse antibody therapy and imaging, and interfering in immunological assays. To investigate the HAMA response, 20 stable cell lines secreting human monoclonal antibodies reactive with mouse IgG were established from a donor with a history of exposure to mice. Their subclass and domain specificities were established by solid-phase binding, indirect haemagglutination assays and immunoblotting, using Igs of known subclass and Ig fragments. The heavy-chain variable region gene usage was determined for 12 HAMAs. Eight HAMAs were IgM, 11 HAMAs were IgG4 and one HAMA was IgG1, indicating an IgG4-dominated response. All of the IgG HAMAs reacted with epitopes present on the Fc portion; one was subclass-specific, nine were subclass restricted and two were pan-IgG-reactive. Measurement of their affinities gave dissociation constants typically in the nanomolar range. Seven and five HAMAs were derived from variable heavy-chain 3 (VH3) and VH1 gene segments, respectively. The IgG HAMAs used different VH segments to the IgM HAMAs. JH regions were coded by JH4 in eight HAMAs. DH segment usage appeared to be restricted in the IgM HAMAs. Two IgG HAMAs were clonally related. These monoclonal HAMAs are potentially useful as reagents for detecting mouse IgG and as reference reagents for the investigation of the HAMA response in patients undergoing mouse monoclonal antibody therapy and for the investigation of the influence of HAMAs on immunodiagnostic tests. PMID- 12542804 TI - Sticky issue of adherence. AB - The phenomenon of adherence, also known as compliance, is the vital link that allows effective medications to have the desired clinical effect when self administered. It is often assumed that the population is generally adherent, but more than 50% of people with chronic illness do not take their medication as prescribed. We highlight how the terminology and language of non-adherence act to conceptualize adherence as a patient problem in a manner that is inadvertently judgmental, narrowly focused and clinically unhelpful. In contrast, knowledge of the dynamic nature of adherence promotes the conceptualization of adherence as the common problem that it is, where the responsibility for improving it lies primarily with the health professional. The example of asthma is used to highlight how individually focused clinical strategies can fit within a population perspective that, in its entirety, can be conceptualized as a framework of adherence-promoting strategies. PMID- 12542805 TI - Rehabilitation of spasticity and related problems in childhood cerebral palsy. AB - Spasticity is one part of the upper motor neuron syndrome, but it is a widespread problem in cerebral palsy; it is debilitating, affects function and can lead to musculoskeletal complications. Significant advances have occurred in antispasticity management (and related musculoskeletal problems) in children with cerebral palsy during the past 5-10 years. Botulinum toxin A has been the most outstanding treatment advance; it is relatively long-lasting, easy to administer, reversible, has a favourable side-effect profile and is highly useful for focal spasticity. There is an emerging role for intrathecal baclofen in Australia. Despite being available and practised in North America for years, selective dorsal rhizotomy has not been popular in Australia. The use of orthopaedic surgery has significantly altered in recent years. There is still a place for oral drug treatment, including some newer agents and the potential for combination treatment with other modalities. The role of physical therapy in defining disability, assessing function, undertaking biomechanical assessment and providing mobility aids/casting/orthoses and motor training/stretching exercises is critical for the success of medical and surgical interventions. From an Australian perspective, the purpose of the present review is to provide a critical review of therapies available for spasticity associated with childhood cerebral palsy. PMID- 12542806 TI - Health-service use by children with asthma over a 6-month period. AB - OBJECTIVES: The present study aims to describe the use of health services by children with asthma, and examine disease-specific, parental and sociodemographic variables associated with different levels of health-service utilization. METHODS: Parents of 135 children attending an emergency room (ER) completed questionnaires measuring the children's asthma symptoms, and sociodemographic and psychological variables. Parents were contacted monthly for 6 months to document the number of planned and unplanned visits to hospital and community health-care services for asthma. RESULTS: At least one further unplanned visit to the ER was made by 37% of children, while 62% made at least one unplanned visit to a general practitioner (GP). Fifty-five per cent made planned review visits to a GP, 30% to paediatricians and 5% to hospital clinics. After controlling for the level of asthma symptoms, parental anxiety and parental perceptions of children's vulnerability were associated with unplanned GP visits (P = 0.05 and P = 0.01, respectively); a planned review visit and the child being admitted to hospital for the index attack were associated with unplanned ER visits (P = 0.05 and P = 0.004, respectively). CONCLUSIONS: Children with asthma more frequently attend GP services than hospital services for both planned and unplanned asthma management. Different variables predict the unplanned use of GP and ER services. Understanding these differences is imperative if children and families are to make the most effective use of health services. PMID- 12542807 TI - Acute flaccid paralysis in Australian children. AB - OBJECTIVES: To describe the epidemiology and causes of acute flaccid paralysis (AFP) in Australian children, and the clinical features of the two most common causes of AFP, Guillain-Barre syndrome and transverse myelitis. METHODS: Monthly active surveillance for AFP was carried out through the Australian Paediatric Surveillance Unit, with AFP defined as 'acute onset of flaccid paralysis in one or more limbs or of bulbar paralysis in any child less than 15 years of age'. RESULTS: Between March 1995 and December 1999, 143 cases of AFP were reported (approximately 0.8 per 100000 children < 15 years of age per annum). The age range was 2 months-14 years and 59% were boys. Out of these children, 137 (96%) were hospitalized and 47 required intensive care. No case of wild or vaccine associated poliomyelitis was identified. The most common causes of AFP were Guillain-Barre syndrome in 67 (47%) and transverse myelitis in 27 (19%). Other diagnoses included acute disseminated encephalomyelitis, trauma, tick-bite paralysis and infantile botulism. CONCLUSION: The participation of paediatricians in AFP surveillance contributed to the accreditation of Australia (along with the other 36 countries of the western Pacific region) as 'polio free' by the World Health Organization in October 2000. The surveillance also provided data on the frequency of AFP and identified Guillain-Barre syndrome and transverse myelitis as the most common diagnoses. In this large national series, many other conditions that may present as non-polio AFP were identified. PMID- 12542808 TI - Idiopathic pulmonary haemosiderosis: an Oriental experience. AB - OBJECTIVES: Idiopathic pulmonary haemosiderosis (IPH) is a rare but potentially lethal disorder. A retrospective analysis of documented cases of IPH in our hospital was conducted in order to study the clinical spectrum and radiographic features, and to explore therapeutic strategies. METHODS: A retrospective chart review was carried out, collecting medical records of patients with pulmonary haemo-siderosis at Chang Gung Children's Hospital (CGCH), a tertiary children's hospital in northern Taiwan. Secondary causes of pulmonary haemosiderosis were excluded. RESULTS: Five patients were diagnosed as having IPH over a 25-year period. The classical triad of IPH was found at initial presentation in only 2/5 patients. One patient had well-established pulmonary fibrosis, but no pulmonary symptoms. The clinical course of IPH was exceedingly variable, with a mean delay of 9 months before diagnosis was made. Bronchoalveolar lavage (BAL) confirmed IPH in 3/5 patients. Immunological abnormalities were noted in two patients, without progression to immune disorders during follow up. While using corticosteroids alone, 4/5 patients continued to have recurrent bleeding episodes. All five patients required immunosuppressive therapy for maintenance of a symptom-free period, but survived to a mean follow up of 2 years. CONCLUSIONS: Early definitive diagnosis and aggressive immunosuppressive therapy of IPH are imperative in order to avoid pulmonary fibrosis and mortality in IPH. A chest radiograph should be included in a serial work-up of unexplained anaemia in children. An examination using BAL can confirm IPH and high-resolution thoracic computed tomography scans are useful for early detection of pulmonary fibrosis. PMID- 12542809 TI - Fatal unintentional traumatic asphyxia in childhood. AB - OBJECTIVE: The present study was undertaken to examine specific features of unintentional traumatic asphyxial deaths in childhood. METHODS: Coronial files and records at the Forensic Science Centre in Adelaide, South Australia, were examined over a 35-year period from 1966 to 2000 for all cases of traumatic asphyxial death occurring in children under the age of 17 years. RESULTS: Six cases of unintentional fatal traumatic asphyxia were identified. All of the victims were boys with an age range of 2-15 years (mean 6.8 years) and all were found dead at the scene. Fatal traumatic asphyxia resulted from entrapment beneath a chest of drawers, beneath a table tennis table, between a pile of wooden pallets and a metal fence, between a conveyor belt and its frame, and under a motor vehicle (in two cases). CONCLUSIONS: Fatal traumatic asphyxia in childhood is a rare event, with younger children commonly being trapped by furniture or by industrial equipment while playing, and older children being trapped under motor vehicles in similar circumstances to adult traumatic asphyxial deaths. Unsupervised play of young children around heavy and potentially unstable pieces of furniture may be dangerous, particularly if more than one child is present. Unsupervised play of young children in industrial yards should be avoided. PMID- 12542810 TI - Comparison of health-risk behaviours among students in alternative high schools from New Zealand and the USA. AB - OBJECTIVES: To describe the prevalence of health-risk behaviours of New Zealand alternative education students, and to compare these behaviours to similar students in the USA. METHODS: Thirty-six alternative education schools in the northern region of New Zealand were surveyed. A total of 269 students completed a youth health questionnaire using laptop computers. These data were compared to data from an equivalent population of alternative education students in the USA. RESULTS: Alternative education students from New Zealand and the USA engage in similar high levels of health-risk behaviours. Female students in New Zealand are at particularly high risk of poor health and social outcomes due to high levels of alcohol and marijuana use, driving under the influence of alcohol and high prevalence of risky sexual behaviours. Such health-risk behaviours place alternative education students at greater risk of some of the leading causes of morbidity and mortality in both youth and adult populations. CONCLUSIONS: Findings from the present study support the need for specific health policies and programmes for alternative high school students. Providers of New Zealand alternative education should be aware that female students are at particularly high risk of many health-risk behaviours. PMID- 12542811 TI - Variations in bronchiolitis management between five New Zealand hospitals: can we do better? AB - OBJECTIVES: To determine the current management of bronchiolitis by five major New Zealand hospitals and to identify areas for improvement. METHODS: Lists of infants under 1 year of age admitted with bronchiolitis during 1998 were obtained from the casemix offices of the five largest New Zealand hospitals with paediatric services. Hospital records from a random sample of these admissions were reviewed. RESULTS: Out of the 409 infants admitted overnight, 8% had been born less than or=32 weeks gestation and 53% were aged younger than 6 months. Overall, 59% received oxygen, 21% had nasogastric fluids, 22% had intravenous fluids, 34% were prescribed antibiotics, 42% received bronchodilators and 60% had a chest radiograph. Respiratory secretions were collected for viral studies from 58% of infants and, in 59%, respiratory syncytial virus was detected. Significant variations in management were detected between hospitals. The overall proportion of infants requiring oxygen, intravenous or nasogastric fluids (65%) was significantly higher than that found in a 1986-1988 Christchurch study where only 25% received one or more of these interventions (P < 0.001). CONCLUSIONS: Opportunities exist to rationalize bronchiolitis management in New Zealand with potential cost savings, particularly by reducing the number of chest radiographs and prescribing of unnecessary antibiotics and bronchodilators. PMID- 12542812 TI - Death due to electrocution in childhood and early adolescence. AB - OBJECTIVES: To delineate the clinicopathological features of fatal childhood electrocutions and to identify specific risk factors. METHODS: Coronial files in Adelaide (Australia) were searched from 1967 to 2001 and Medical Examiners' files in San Diego (USA) were searched from 1988 to 2001, for cases of deaths of children and adolescents younger than 16 years attributed to electrocution. RESULTS: Sixteen cases were identified aged between 10 months and 15 years (mean 8.0 years) with a male : female ratio of 5 : 3. Deaths were due to accidents occurring while playing with or near faulty electrical equipment at home or at school (n = 8), electrical equipment while in the bath (n = 2), damaged outdoor electrical equipment (n = 1), overhead wires (n = 1), and a high voltage electricity substation (n = 1). In addition, one death was due to suicide involving an electrical appliance placed in a bath, and two other deaths occurred in older children who were moving equipment under overhead wires. No homicides were identified. CONCLUSIONS: Childhood deaths due to electrocution are rare and are more likely to occur when children are playing around electrical wires or equipment, and often result from either faulty apparatus, or a lack of understanding of the potential dangers involved. The majority of deaths (11/16; 69%) occur in the home environment. In contrast to adult electrical deaths, high voltage electrocutions, suicides and workplace deaths are uncommon. Strategies for eliminating childhood electrocution should concentrate on ensuring safe domestic environments with properly maintained electrical devices. PMID- 12542813 TI - Parents' Evaluation of Developmental Status in the Australian day-care setting: developmental concerns of parents and carers. AB - OBJECTIVE: To trial the Parents' Evaluation of Developmental Status (PEDS) as an instrument for reporting developmental concerns for Australian preschool children. METHOD: A cross-sectional survey of parents and carers of 262 children attending five day-care centres and two kindergartens in Melbourne was conducted between October and November 1999. Parents and carers completed the written PEDS questionnaire, comprising 10 questions eliciting concerns about learning, development and behaviour, and answered questions about the acceptability and use of the PEDS. RESULTS: Of 445 children, 389 were eligible for inclusion. Complete parent and carer PEDS data were available for 262 children (67% response: 47% boys; 53% girls) aged from 18 months to 5 years, 9 months. Most parents found the PEDS questionnaire easy to complete (98%) and likely to be useful to health professionals (89%). Twenty-four children (9%) were classified as being at high risk of disabilities and 49 (19%) were classified as being at medium-risk of disabilities. Parents of 125 children (48%) reported no concerns. The prevalence of parental concerns was similar to the USA norming sample. Carers reported similar prevalences of concerns. Although overall agreement was high, parent/carer kappa-values were modest, being highest for the gross motor (kappa = 0.40) and social-emotional (kappa = 0.37) domains. CONCLUSIONS: The PEDS is acceptable to parents of Australian preschool children, with a prevalence of significant concerns (i.e. children at high- and medium-risk of developmental problems) that is similar to those in the USA. Further research is needed to assess what factors differentially influence whether a concern is felt in a particular domain for a particular child. PMID- 12542814 TI - School-aged children with Kawasaki disease: high incidence of cervical lymphadenopathy and coronary artery involvement. AB - OBJECTIVE: We describe 10 school-aged children with Kawasaki disease (KD) with a high incidence of cervical lymphadenopathy and coronary abnormality. METHODS: Based on a database of 1002 children with KD in Chang Gung Children's Hospital from January 1983 to March 2001, 10 (1%) school-aged patients (five boys, five girls) who met the diagnostic criteria of KD were included for analysis. RESULTS: Cervical lymphadenopathy was noted in all (100%) of these patients. Unilateral neck mass mimicking acute suppurative infections not responding to antibiotic therapy was the initial presentation in nine (90%) of the 10 patients. The mean interval between disease onset and diagnosis was 9.9 +/- 3.3 days (range, 6-15 days). Seven (70%) of these patients responded to one course of high-dose intravenous immunoglobulin (IVIG) therapy (2 g/kg) and oral aspirin (80-100 mg/kg per day), two (20%) required a second course of IVIG, and one (10%) responded to high-dose aspirin treatment only. Coronary artery abnormality (dilatation or aneurysm) was documented by echocardiography in seven (70%) patients (four boys, three girls). In six patients, the coronary artery abnormalities resolved in 1 year, while one patient had persistent right coronary artery aneurysm, which necessitated continued anticoagulant and low-dose aspirin therapy. CONCLUSION: The incidence of school-aged children among patients with KD is about 1% in our hospital. These patients are notable for the high incidence of initial manifestations of unilateral neck mass and coronary artery involvement. This disease should be listed as the differential diagnosis in school-aged children presenting with fever and neck mass that do not respond to antibiotic therapy. PMID- 12542815 TI - Moral status of the fetus: fetal rights or maternal autonomy? PMID- 12542816 TI - HIV in pregnancy: interests of the mother and the baby. PMID- 12542819 TI - Blue lips, coma and haemolysis. PMID- 12542820 TI - Polyneuritis cranialis with contrast enhancement of cranial nerves on magnetic resonance imaging. AB - The disorder of multiple cranial nerve palsies without spinal cord involvement is referred to as polyneuritis cranialis (PC) and is rare. It is thought to be an acute post-infective polyneuropathy or a variant of Guillain-Barre syndrome. Electrophysiological evidence of demyelination has been reported, but no radiological abnormalities of the affected cranial nerves have been noted. We report a case of PC where contrast enhanced magnetic resonance imaging (MRI) showed enhancement of the peripheral segments of the oculomotor and abducens nerves. This case illustrates the utility of MRI in the assessment of cranial nerve palsies. PMID- 12542821 TI - Iatrogenic injury in childhood staphylococcal scalded skin syndrome. AB - Staphylococcal scalded skin syndrome (SSSS) may cause significant morbidity in children. It is common practice for adhesive occlusive dressings to be used to apply topical local anaesthetic prior to venepuncture. We report two cases in which removal of these dressings from children with SSSS caused injury and discomfort in areas previously free from blistering. We recommend that an alternative method of topical anaesthetic application is used to minimize skin trauma in these patients. PMID- 12542822 TI - Scurvy: forgotten but not gone. AB - Scurvy is still seen sporadically in the developed world. At a time when subclinical vitamin C deficiency in the general population is being recognized increasingly, the need for clinicians to be aware of this disease remains. We present the case of a 9-year-old boy admitted to hospital with musculoskeletal pain, weakness and changes in the skin and gums. After extensive investigation, he was found to have vitamin C deficiency resulting from a restricted eating pattern. Musculoskeletal complaints are a common mode of presentation of scurvy in children. Failure to appreciate this fact and the risk factors for poor vitamin C intake in the paediatric age group can result in unnecessary and invasive investigations for apparent 'multisystem' disease. PMID- 12542827 TI - Which putatively pre-malignant oral lesions become oral cancers? Clinical relevance of early targeting of high-risk individuals. AB - Oral squamous cell carcinomas continue to be a group of diseases with high mortality and increasing incidence rates, particularly among young individuals. This is a paradox finding, since most oral cancers are preceded - even by several years - by readily detectable mucosal changes, most often white or red patches (leukoplakias and erythroplakias, respectively). However, only a small fraction of leukoplakias or erythroplakias are related to cancer development, and the challenge has been to identify the high-risk lesions. From the vast literature on molecular markers in oral pre-cancer, no reliable prognostic marker for risk assessment in putatively pre-malignant lesions has emerged. For this reason, a wait-and-see approach has been adopted for this group of lesions. Recently published data point to gross genomic aberrations (DNA aneuploidy) as a tool for targeting patients at particular risk for future cancerous lesions in the oral cavity. Thus, DNA aneuploidy signalled a very high risk for subsequent development of carcinomas in a wide range of lesions from the oral mucous membrane, ranging from oral leukoplakias to oral erythroplakias and even including lesions that had been explicitly defined as being without a malignant potential by a group of trained pathologists. As an extension of research in oral pre-malignancies, the enzyme cyclooxygenase-2 (COX-2) seems to be enhanced specifically in high-risk oral lesions, as defined by the aberrant DNA content of their lesions. These data strongly indicate that COX-2 inhibitors (coxibs) should be investigated as chemopreventive agents in patients identified to be at high risk of developing oral cancer. PMID- 12542828 TI - Usefulness of AgNOR counts in diagnosing epithelial dysplasia. AB - BACKGROUND: Diagnosis of epithelial dysplasia has traditionally been subjective, and there is a need for a quantifiable and useful test. METHODS: In a double blind study, clinical leukoplakias from 52 people were diagnosed for presence (DLK) or absence (NDLK) of epithelial dysplasia using hematoxylin and eosin (H&E) stain as a gold standard criterion, and results were compared against their mean silver stainable nucleolar organizer region (AgNOR) counts. We used mean AgNOR count cut-point of 2.37 from our prior report as the diagnostic threshold (mean >or= 2.37 being DLK and mean < 2.37 being NDLK). RESULTS: The two methods (H&E and AgNOR) disagreed in 37% of the diagnoses. Both NDLK and DLK had high AgNOR counts. P-AgNOR was non-contributory for diagnosing epithelial dysplasia. CONCLUSIONS: Mean AgNOR count can be a useful tool in definitive diagnosis of epithelial dysplasia. PMID- 12542829 TI - Th1 cytokines in oral lichen planus. AB - BACKGROUND: Cell-mediated immune responses in oral lichen planus (OLP) may be regulated by cytokines and their receptors. METHODS: In situ cytokine expression and in vitro cytokine secretion in OLP were determined by immunohistochemistry and ELISA. RESULTS: The majority of subepithelial and intraepithelial mononuclear cells in OLP were CD8+. In some cases, intraepithelial CD8+ cells were adjacent to degenerating keratinocytes. CD4+ cells were observed mainly in the deep lamina propria with occasional CD4+ cells close to basal keratinocytes. Mononuclear cells expressed IFN-gamma in the superficial lamina propria and TNF-alpha adjacent to basal keratinocytes. Basal keratinocytes expressed TNF-alpha as a continuous band. TNF R1 was expressed by mononuclear cells and basal and suprabasal keratinocytes. There was variable expression of TGF-beta1 in the subepithelial infiltrate while all intraepithelial mononuclear cells were TGF beta1-. Keratinocytes in OLP stained weakly for TGF-beta1. Unstimulated OLP lesional T cells secreted IFN-gamma in vitro. TNF-alpha stimulation down regulated IFN-gamma secretion and up-regulated TNF-alpha secretion. IL-4, IL-10 and TGF-beta1 secretion were not detected. CONCLUSIONS: These data suggest the development of a T helper 1 immune response that may promote CD8+ cytotoxic T cell activity in OLP. PMID- 12542830 TI - Hepatocyte growth factor and c-Met (HGF/c-Met) in adenoid cystic carcinoma of the human salivary gland. AB - BACKGROUND: Hepatocyte growth factor (HGF) enhances cell growth, morphogenesis, and scattering of various epithelial cells. The aim of this study was to evaluate the hypothesis that HGF/c-Met plays a biological role in the invasive growth of adenoid cystic carcinoma (ACC). METHODS: Immunohistochemically, expression of HGF and its receptor c-Met was examined in 15 cases of ACC. To examine the direct effects of HGF on ACC, cell line derived from of ACC (ACC3) was used. The expression of HGF and c-met in ACC3 was investigated by RT-PCR. Analysis of mechanisms of invasion was done by performing scattering assay and matrigel invasion chamber assay. RESULTS: Positive staining of HGF was found in all cases, and that of c-Met was 67%. In ACC3, c-met was expressed, but not HGF. Stimulation of ACC3 by rhHGF induced scattering and promoted invasion. CONCLUSION: The present results suggest that HGF/c-Met increases tumor cell scattering and may play a part in invasiveness of ACC. PMID- 12542831 TI - Proliferation and distribution of myoepithelial cells during atrophy of the rat sublingual gland. AB - BACKGROUND: The present study was aimed to determine the proliferation and distribution of myoepithelial cells during atrophy of rat sublingual glands. METHODS: The excretory duct of the right sublingual gland of rats was doubly ligated with metal clips to induce atrophy in the gland. The atrophic sublingual glands were taken from 1 to 28 days after duct ligation and examined with single immunohistochemistry for actin as a marker of myoepithelial cells and with immunohistochemical double staining for actin and proliferating cell nuclear antigen (PCNA) as a marker of proliferating cells. RESULTS: In unligated sublingual glands, myoepithelial cells embraced acini and intercalated ducts, but not striated and interlobular excretory ducts. In the early stages of atrophy, myoepithelial cells surrounded small ducts but not large ones. However, in the later stages of atrophy, myoepithelial cells were also observed at the periphery of the large ducts. The immunohistochemical double staining showed that there were PCNA-positive myoepithelial cells in the normal as well as in the atrophic sublingual glands. However, the PCNA labeling indices of myoepithelial cells were low in the unligated and atrophic sublingual glands, and there were no statistically significant differences in these labeling indices. CONCLUSION: The observations suggest that the distribution of myoepithelial cells change during atrophy of rat sublingual glands and that myoepithelial cells have low proliferative activity in both the normal and atrophic condition of rat sublingual glands. PMID- 12542832 TI - Alterations of nitrate and nitrite content in saliva, serum, and urine in patients with salivary dysfunction. AB - BACKGROUND: The role of the salivary glands in the maintenance of nitrate and nitrite in saliva is poorly understood. The aim was to study alterations of nitrate and nitrite metabolisms in patients with Sjogren's syndrome (SS) or sialosis. METHODS: Saliva, serum, and urine samples were collected from healthy volunteers (n = 29), patients with SS (n = 31), and patients with sialosis (n = 30). Concentrations of nitrate and nitrite were determined by high-performance liquid chromatography (HPLC). RESULTS: In the healthy group, the highest concentration of nitrate was found in parotid saliva (172 mg/l), followed by urine (160 mg/l), whole saliva (97 mg/l), and serum (33 mg/l). In the SS group, concentration of nitrate was decreased in parotid saliva and whole saliva, and increased significantly in urine. Concentration of nitrite in whole saliva was significantly decreased in the SS group and increased in the sialosis group. CONCLUSION: Hypofunction of the salivary glands is associated with significant changes of nitrate and nitrite levels in the saliva and urine. PMID- 12542833 TI - Laminin-5 gamma 2 chain is colocalized with gelatinase-A (MMP-2) and collagenase 3 (MMP-13) in odontogenic keratocysts. AB - BACKGROUND: Odontogenic keratocyst (KC) differs from other epithelial odontogenic cysts in regard to increased epithelial proliferation and a strong tendency to recur. Laminin-5 (Ln-5) is an epithelial anchoring filament component, which after modulation by certain matrix metalloproteinases (MMPs), like MMP-2 and MMP 13, induces epithelial cell migration. METHODS: Using in situ hybridization and immunohistochemistry, we studied the Ln-5 gamma-2 chain expression related to the expression of MMP-2, -8, and -13 in different odontogenic cysts, including radicular cysts (RC; n = 11), follicular cysts (FC; n = 11), and odontogenic keratocysts (KC; n = 16). RESULTS: Ln-5 mRNA was present in all cysts examined, while less than half of KCs and RCs (33 and 40%, respectively) demonstrated MMP-2 mRNA. MMP-13 mRNA was present in all KC samples. Ln-5 protein was located as a continuous ribbon in BM zone of all KCs, and MMP-2 and MMP-13 immunoreactivities colocated significantly with Ln-5 in that area. MMP-8 was expressed by stromal macrophages and epithelial goblet cells, but never located in BM zone. CONCLUSIONS: Our results indicate that the colocalization of Ln-5 with MMP-2 or MMP-13, but not with MMP-8, in BM zone of KCs, may be related to special characteristics of KC. PMID- 12542834 TI - Immunohistochemical and genetic analysis of mandibular cysts in heterozygous ptc knockout mice. AB - BACKGROUND: Alterations of human patched (ptc) homolog have been proven to be responsible for basal cell nevus syndrome (BCNS). Mandibular cysts in heterozygous ptc knockout mouse (ptc+/- mouse) were microradiologically, histologically, immunohistochemically, and genetically examined to investigate the possible role of the ptc gene and its associates in the jaw cysts. METHODS: The mandibular bones were prepared from 63 ptc+/- mice and 6 ptc+/+ mice. Soft X ray radiographs and histological sections were examined for detection of the presence of mandibular cysts. The mandibular cysts were immunohistochemically investigated using anti-ptc, shh, and smo antibodies. PCR analysis of loss of heterozygosity (LOH) of ptc was performed in genomic DNA from the mandibular cysts. RESULTS: Six ptc+/+ mice showed no pathologic change in any examinations. Microradiologically, ptc+/- mice did not show any apparent lesion. Mandibular cysts were often multiple, and were histologically detected in the alveolar bones or periodontal ligaments of the molars in 16 (25.4%) ptc+/- mice. The mandibular cysts were lined by thin parakeratotic stratified squamous epithelium and contained keratinized materials. Immunohistochemical examination showed sonic hedgehog (shh) protein mainly in cyst lining epithelium, and ptc and smoothened (smo) proteins in cyst lining epithelium, and surrounding fibrous connective tissue. Expression of ptc protein in the cyst lining epithelium tended to be weak as compared with incisor enamel organs and gingival stratified squamous epithelium. LOH of the ptc gene couldn't be found in lining epithelium of mandibular cysts in any ptc+/- mice. CONCLUSIONS: Ptc+/- mouse is a useful model of BCNS from the standpoint of occurrence of jaw cysts, and downregulation of ptc protein in cyst lining epithelium caused by gene targeting would be associated with formation of jaw cysts in ptc+/- mice. PMID- 12542835 TI - Immunohistochemical detection of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in ameloblastomas. AB - BACKGROUND: To evaluate the roles of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in tumor progression, expression of MMP-1, -2 and -9 and TIMP-1 and -2 was analyzed in ameloblastomas as well as tooth germs. METHODS: Frozen tissue sections of seven tooth germs and 22 ameloblastomas were immunohistochemically examined using anti-MMP-1, -2 and -9 and anti-TIMP-1 and -2 antibodies. RESULTS: MMP-1, -2 and -9 and TIMP-1 and -2 were expressed strongly in mesenchymal components of tooth germs, and stromal cells of ameloblastomas. Immunoreactivity for MMP-9 in stromal cells of ameloblastomas was significantly stronger than in mesenchymal cells of dental follicles and dental papillae. Dental laminae showed weak MMP-2 expression in six tooth germs, MMP-9 expression in two tooth germs and TIMP-1 expression in six tooth germs. Some tumor cells showed weak MMP-2 expression in 19 ameloblastomas, MMP-9 expression in four ameloblastomas and TIMP-1 expression in all cases. TIMP-2 reactivity was prominently found in basement membrane zones of dental laminae in tooth germs, and tumor cell islands or nests in ameloblastomas. CONCLUSION: Expression of MMPs and TIMPs was considered to be associated with interactions between epithelial cells and mesenchymal components in normal and neoplastic odontogenic tissues; these molecules might play a role in regulation of tumor progression in ameloblastomas as well as regulation of developmental processes in tooth germs. PMID- 12542836 TI - A case of sebaceous carcinoma of the parotid gland. AB - Sebaceous carcinoma of salivary gland origin is an extremely rare malignancy. It occurs mainly in the parotid gland. This is a case report of a sebaceous carcinoma in a 57-year-old woman who had a lump over the right parotid region for 8-9 months. The tumour was composed of small basaloid cells and large foamy cells. Sebaceous differentiation was evident in some tumour islands. This is the first case reported in the Department of Oral Pathology, Faculty of Dental Sciences, University of Peradeniya. PMID- 12542838 TI - Time series modeling for syndromic surveillance. AB - BACKGROUND: Emergency department (ED) based syndromic surveillance systems identify abnormally high visit rates that may be an early signal of a bioterrorist attack. For example, an anthrax outbreak might first be detectable as an unusual increase in the number of patients reporting to the ED with respiratory symptoms. Reliably identifying these abnormal visit patterns requires a good understanding of the normal patterns of healthcare usage. Unfortunately, systematic methods for determining the expected number of (ED) visits on a particular day have not yet been well established. We present here a generalized methodology for developing models of expected ED visit rates. METHODS: Using time series methods, we developed robust models of ED utilization for the purpose of defining expected visit rates. The models were based on nearly a decade of historical data at a major metropolitan academic, tertiary care pediatric emergency department. The historical data were fit using trimmed-mean seasonal models, and additional models were fit with autoregressive integrated moving average (ARIMA) residuals to account for recent trends in the data. The detection capabilities of the model were tested with simulated outbreaks. RESULTS: Models were built both for overall visits and for respiratory-related visits, classified according to the chief complaint recorded at the beginning of each visit. The mean absolute percentage error of the ARIMA models was 9.37% for overall visits and 27.54% for respiratory visits. A simple detection system based on the ARIMA model of overall visits was able to detect 7-day-long simulated outbreaks of 30 visits per day with 100% sensitivity and 97% specificity. Sensitivity decreased with outbreak size, dropping to 94% for outbreaks of 20 visits per day, and 57% for 10 visits per day, all while maintaining a 97% benchmark specificity. CONCLUSIONS: Time series methods applied to historical ED utilization data are an important tool for syndromic surveillance. Accurate forecasting of emergency department total utilization as well as the rates of particular syndromes is possible. The multiple models in the system account for both long-term and recent trends, and an integrated alarms strategy combining these two perspectives may provide a more complete picture to public health authorities. The systematic methodology described here can be generalized to other healthcare settings to develop automated surveillance systems capable of detecting anomalies in disease patterns and healthcare utilization. PMID- 12542839 TI - The role of strategy and redundancy in diagnostic reasoning. AB - BACKGROUND: Diagnostic reasoning is a key competence of physicians. We explored the effects of knowledge, practice and additional clinical information on strategy, redundancy and accuracy of diagnosing a peripheral neurological defect in the hand based on sensory examination. METHOD: Using an interactive computer simulation that includes 21 unique cases with seven sensory loss patterns and either concordant, neutral or discordant textual information, 21 3rd year medical students, 21 6th year and 21 senior neurology residents each examined 15 cases over the course of one session. An additional 23 psychology students examined 24 cases over two sessions, 12 cases per session. Subjects also took a seven-item MCQ exam of seven classical patterns presented visually. RESULTS: Knowledge of sensory patterns and diagnostic accuracy are highly correlated within groups (R2 = 0.64). The total amount of information gathered for incorrect diagnoses is no lower than that for correct diagnoses. Residents require significantly fewer tests than either psychology or 6th year students, who in turn require fewer than the 3rd year students (p < 0.001). The diagnostic accuracy of subjects is affected both by level of training (p < 0.001) and concordance of clinical information (p < 0.001). For discordant cases, refutation testing occurs significantly in 6th year students (p < 0.001) and residents (p < 0.01), but not in psychology or 3rd year students. Conversely, there is a stable 55% excess of confirmatory testing, independent of training or concordance. CONCLUSIONS: Knowledge and practice are both important for diagnostic success. For complex diagnostic situations reasoning components employing redundancy seem more essential than those using strategy. PMID- 12542840 TI - Implementing guidelines in primary care: can population impact measures help? AB - BACKGROUND: Primary care organisations are faced with implementing a large number of guideline recommendations. We present methods by which the number of eligible patients requiring treatment, and the relative benefits to the whole population served by a general practice or Primary Care Trust, can be calculated to help prioritise between different guideline recommendations. METHODS: We have developed measures of population impact, "Number to be Treated in your Population (NTP)" and "Number of Events Prevented in your Population (NEPP)". Using literature-based estimates, we have applied these measures to guidelines for pharmacological methods of secondary prevention of myocardial infarction (MI) for a hypothetical general practice population of 10,000. RESULTS: Implementation of the NICE guidelines for the secondary prevention of MI will require 176 patients to be treated with aspirin, 147 patients with beta-blockers and with ACE Inhibitors and 157 patients with statins (NTP). The benefit expressed as NEPP will range from 1.91 to 2.96 deaths prevented per year for aspirin and statins respectively. The drug cost per year varies from euro 1940 for aspirin to euro 60,525 for statins. Assuming incremental changes only (for those not already on treatment), aspirin post MI will be added for 37 patients and produce 0.40 of a death prevented per year at a drug cost of euro 410 and statins will be added for 120 patients and prevent 2.26 deaths per year at a drug cost of euro 46,150. An appropriate policy might be to reserve the use of statins until eligible patients have been established on aspirin, ACE-Inhibitors and beta blockers. CONCLUSIONS: The use of population impact measures could help the Primary Care Organisation to prioritise resource allocation, although the results will vary according to local conditions which should be taken into account before the measures are used in practice. PMID- 12542841 TI - Effect of human leukocyte antigen heterozygosity on infectious disease outcome: the need for allele-specific measures. AB - BACKGROUND: Doherty and Zinkernagel, who discovered that antigen presentation is restricted by the major histocompatibility complex (MHC, called HLA in humans), hypothesized that individuals heterozygous at particular MHC loci might be more resistant to particular infectious diseases than the corresponding homozygotes because heterozygotes could present a wider repertoire of antigens. The superiority of heterozygotes over either corresponding homozygote, which we term allele-specific overdominance, is of direct biological interest for understanding the mechanisms of immune response; it is also a leading explanation for the observation that MHC loci are extremely polymorphic and that these polymorphisms have been maintained through extremely long evolutionary periods. Recent studies have shown that in particular viral infections, heterozygosity at HLA loci was associated with a favorable disease outcome, and such findings have been interpreted as supporting the allele-specific overdominance hypothesis in humans. METHODS: An algebraic model is used to define the expected population-wide findings of an epidemiologic study of HLA heterozygosity and disease outcome as a function of allele-specific effects and population genetic parameters of the study population. RESULTS: We show that overrepresentation of HLA heterozygotes among individuals with favorable disease outcomes (which we term population heterozygote advantage) need not indicate allele-specific overdominance. On the contrary, partly due to a form of confounding by allele frequencies, population heterozygote advantage can occur under a very wide range of assumptions about the relationship between homozygote risk and heterozygote risk. In certain extreme cases, population heterozygote advantage can occur even when every heterozygote is at greater risk of being a case than either corresponding homozygote. CONCLUSION: To demonstrate allele-specific overdominance for specific infections in human populations, improved analytic tools and/or larger studies (or studies in populations with limited HLA diversity) are necessary. PMID- 12542843 TI - Tetracycline-regulated gene expression in replication-incompetent herpes simplex virus vectors. AB - Although herpes simplex virus (HSV) vectors appear to have great potential as gene delivery vectors both in vitro and in vivo, the expression of foreign genes in such vectors cannot be easily regulated. Of the known eukaryotic regulatory systems, the tetracycline-inducible gene expression system is perhaps the most widely used because of its induction characteristics and because of the well known pharmacological properties of tetracycline (Tet) and analogs such as doxycycline. Here, we describe the adaptation of the Tet-inducible system for use in replication-incompetent HSV vectors. HSV vectors were constructed that contained several types of Tet-inducible promoters for foreign gene expression. These promoters contained a tetracycline response element (TRE) linked to either a minimal cytomegalovirus (CMV) immediate-early promoter, a minimal HSV ICP0 promoter, or a truncated HSV ICP0 promoter containing one copy of the HSV TAATGARAT cis-acting immediate-early regulatory element (where R represents a prime base). All three promoter constructs were regulated appropriately by doxycycline, as shown by the expression of the marker gene lacZ in cell lines engineered to express Tet transactivators. The ICP0 promoter constructs expressed the highest and most sustained levels of lacZ, but the CMV promoter construct had the highest relative level of induction, suggesting their use in different applications. To extend the utility of Tet-regulated HSV vectors, vectors were constructed that coexpressed an inducible Tet transactivator in addition to the inducible lacZ marker gene. This modification resulted in tetracycline-inducible gene expression that was not restricted to specific cell lines, and this vector was capable of inducible expression in irreversibly differentiated NT2 cells (NT neurons) for several days. Finally, HSV vectors were constructed that expressed modified Tet transactivators, resulting in improved induction properties and indicating the flexibility of the Tet-regulated system for regulation of foreign gene expression in HSV vector-infected cells. PMID- 12542842 TI - In silico characterisation and chromosomal localisation of human RRH (peropsin)- implications for opsin evolution. AB - BACKGROUND: The vertebrate opsins are proteins which utilise a retinaldehyde chromophore in their photosensory or photoisomerase roles in the visual/irradiance detection cycle. The majority of the opsins, such as rod and cone opsins, have a very highly conserved gene structure suggesting a common lineage. Exceptions to this are RGR-opsin and melanopsin, whose genes have very different intron insertion positions. The gene structure of another opsin, peropsin (retinal pigment epithelium-derived rhodopsin homologue, RRH) is unknown. RESULTS: By in silico analysis of the GenBank database we have determined that the human RRH comprises 7 exons spanning approximately 16.5 kb and is localised to chromosome 4q25 in the following gene sequence: cen-EGF-RRH IF-qter - a position that excludes this gene as a candidate for the RP29 autosomal recessive retinitis pigmentosa locus. A comparison of opsin gene structures reveals that RRH and RGR share two common intron (introns 1 and 4) insertion positions which may reflect a shared ancestral gene. CONCLUSION: The opsins comprise a diverse group of genes which appear to have arisen from three different lineages. These lineages comprise the "classical opsin superfamily" which includes the rod and cone opsins, pinopsin, VA-opsin, parapinopsin and encephalopsin; the RRH and RGR group; and the melanopsin line. A common lineage for RRH and RGR, together with their sites of expression in the RPE, indicates that peropsin may act as a retinal isomerase. PMID- 12542844 TI - Glucose-regulated glucose uptake by transplanted muscle cells expressing glucokinase counteracts diabetic hyperglycemia. AB - Type 1 diabetic patients depend on insulin replacement therapy. However, chronic hyperglycemia due to failure to maintain proper glycemic control leads to microvascular, macrovascular, and neurological complications. Increased glucose disposal by tissues engineered to overexpress key regulatory genes in glucose transport or phosphorylation can reduce diabetic hyperglycemia. Here we report that differentiated myoblast cells expressing the glucose-phosphorylating enzyme glucokinase (GK) showed a glucose-dependent increase in glucose uptake and utilization in vitro. Transplantation of GK-expressing myotubes into healthy mice did not alter blood glucose levels and recipient mice maintained normoglycemia. After streptozotocin treatment, mice transplanted with GK-expressing myotubes counteracted hyperglycemia, polydipsia, and polyphagia, whereas mice transplanted with control myotubes developed diabetes. Similarly, diabetic mice transplanted with control myotubes remained hyperglycemic. In contrast, transplantation of GK expressing myotubes into diabetic mice lowered hyperglycemia. These results suggest that the use of genetically engineered muscle cells to express glucokinase may provide a glucose-regulated approach to reduce diabetic hyperglycemia. PMID- 12542845 TI - Production of recombinant H1 parvovirus stocks devoid of replication-competent viruses. AB - Vector and helper plasmids for the production of recombinant H1 (rH1) parvovirus, an oncolytic virus and candidate vector for cancer gene therapy, were constructed with the aim of reducing the contamination of these preparations with replication competent viruses (RCV). Split-helper plasmids were constructed by manipulating the splicing signals for the capsid proteins such that VP1 and VP2 were expressed from separate plasmids. H1 vectors with similarly mutated splice sites were packaged, using the split-helper plasmids, and the resulting recombinant H1 viruses were completely free of RCV because the generation of recombinants expressing both capsid proteins was prevented. Vector yields of rH1 produced with split-helper plasmids in combination with splice site-modified vectors were similar (in the range of 10(7) replication units/ml) to yields of rH1 produced with the standard vector/helper pair, in which case significant levels of RCV were generated (10(4)-10(5) plaque-forming units/ml). To assess the functionality of this approach in vivo, rH1 was produced that contained the human interleukin 2 (IL-2) transgene and that was devoid of RCV. This IL-2-carrying rH1 vector expressed IL-2 efficiently in human tumor cells (HeLa) in vitro and generated antitumor responses in nude mice xenografted with HeLa cells that had been infected ex vivo with this virus. These results should allow the large-scale production of recombinant oncotropic parvoviruses and their assessment for the gene therapy of cancer in a clinical setting. PMID- 12542846 TI - Dynamics of transgene expression in human glioblastoma cells mediated by herpes simplex virus/adeno-associated virus amplicon vectors. AB - One of the challenges in gene therapy is to ensure stable transgene expression at the site of disease with a high degree of accuracy and safety. In this paper, we examine both viral and cellular elements that may affect the level of transgene expression mediated by herpes simplex virus type 1 (HSV-1) adeno-associated virus (AAV) amplicon vectors. These elements include the AAV inverted terminal repeats (ITRs), the AAV Rep proteins, and the allelic status of 19q in human glioma cell lines. The latter is of particular interest because the AAV integration site (AAVS1) is located on the long arm of chromosome 19 and 30-40% of human glioblastoma tumors are reported to have loss of heterozygosity in this region of chromosome 19q. Fluorescence-activated cell-sorting analysis results indicate that inclusion of minimal or full-length AAV ITRs in HSV-1 amplicon vectors markedly increases the efficiency of transgene expression. On the other hand, insertion of the AAV rep gene decreases the level of transgene expression, apparently because of the cytotoxic effects of Rep proteins. Further, the levels of transgene expression appear to be independent of 19q allelic status or the number of endogenous AAVS1 sequences in the various glioma cell lines studied. Taken together, these data support employing AAV ITRs, in the context of HSV-1 amplicon vectors, to enhance short-term levels of transgene expression. PMID- 12542847 TI - Tetracycline transcriptional silencer tightly controls transgene expression after in vivo intramuscular electrotransfer: application to interleukin 10 therapy in experimental arthritis. AB - The doxycycline (Dox)-inducible reverse tetracycline transactivator (rtTA) is often used to control gene expression. However, the Tet-on system displays a high background activity. To overcome this unregulated expression we used the tetracycline-dependent transcriptional silencer (tTS), which binds the tetO inducible promoter in the absence of Dox. Controlled gene expression was analyzed in vivo by delivering combinations of Dox-regulated luciferase reporter construct, rtTA, and tTS expression plasmids into mouse muscle, using electrotransfer. Elevated luciferase expression levels were observed in the absence of doxycycline, and a 10-fold induction was obtained after drug administration. In contrast, when tTS was added, background expression was dramatically lowered by three to four orders of magnitude, and induction was maintained. The tTS system was then used to control expression of a therapeutic gene in experimental arthritis. DBA/1 mice were coinjected with plasmids encoding the antiinflammatory interleukin-10 cytokine under the control of the tetO promoter, the rtTA, and the tTS. Electrotransfer resulted in a dose-dependent increase in IL-10 expression, maintained over a 3-month period, and significant inhibitory effects on collagen-induced arthritis. We conclude that the use of tTS significantly improves the utility of the rtTA system for somatic gene transfer by reducing background activity. PMID- 12542848 TI - Improvement of spatial learning and memory after adenovirus-mediated transfer of the nerve growth factor gene to aged rat brain. AB - Adenovirus-mediated transfer of the nerve growth factor gene promotes significant recovery of age-related cholinergic neuronal deficits in aged rats, but the effects of such treatment on cognitive dysfunction remain unclear. Herein we report a beneficial effect of first-generation adenovirus-mediated nerve growth factor gene transfer (AdNGF) on the spatial learning and memory of aged rats. The NGF protein was detected by enzyme-linked immunosorbent assay in cerebrospinal fluid as early as 3 days after gene transfer and was expressed for at least 30 days. Escape latency in the Morris water maze hidden-platform test was significantly improved on day 8 postinoculation in memory-impaired rats treated with AdNGF as well as at later testing intervals. Ultimately, the escape latency values for the AdNGF group become indistinguishable from those for aged rats with normal learning capacity. Immunohistochemical analysis of septal cholinergic neurons for choline acetyltransferase (ChAT) showed significant increases in both the number and somal distribution of ChAT-positive cells after inoculation of memory-impaired rats with AdNGF. Improvement in memory performance was positively correlated with increases in both NGF concentration in cerebrospinal fluid (r = 0.73, p = 0.005) and the number of ChAT-staining cells (r = 0.77, p = 0.0022). We conclude that AdNGF can improve cognitive function in memory-impaired aged rats and, with refinements in vector-driven expression of the transgene, may prove suitable for use in humans. PMID- 12542849 TI - Preexisting antiadenoviral immunity and regional myocardial gene transfer: modulation by nitric oxide. AB - The utility of adenoviral vectors, currently used in cardiovascular gene transfer protocols, is limited by the brevity of transgene expression and by antiadenoviral immune responses. The effect of preexisting antiadenoviral immunity on intracardiac gene transfer or its modulation by nitric oxide is unknown. Adenoviral vectors, expressing the firefly luciferase gene (AdLuc) or the human nitric oxide synthase 3 (NOS3) gene (AdNOS3), were infused into the great cardiac vein of naive pigs or immunized pigs. Pigs were immunized by intravenous injection of control virus AdRR5 and the resulting neutralizing antibody titers (median, 1:178; p < 0.0001 vs. baseline) were similar to preexisting titers in 54% of randomly selected coronary artery bypass graft patients. In naive animals distribution of transgene expression in the left ventricular free wall was focal. In immunized pigs myocardial luciferase expression 3 days after AdLuc gene transfer was more than 1000-fold lower than in naive pigs, whereas no change in NOS3 transcript levels was detected after AdNOS3 gene transfer. Severe, grade III-IV mononuclear cell infiltration and myocyte apoptosis were observed in four of five AdLuc-infected, immunized animals, compared with low-level inflammation and apoptosis in five of six AdNOS3-infected pigs. Coinfusion of AdLuc and AdNOS3 in immunized pigs resulted in spatially colocalized transgene expression, reduced T cell-mediated inflammation, and myocyte apoptosis and was associated with 200-fold greater median reporter transgene expression levels in the subendocardium (1.0 x 10(3) light units [LU]/mg protein, n = 8, vs. 4.5 x 10(1) LU/mg protein in AdLuc- and AdRR5 coinfected pigs, n = 7, p = 0.02). Preexisting antiadenoviral immunity abrogates myocardial gene expression in pigs and is associated with severe inflammation and myocyte apoptosis. Intracardiac NOS3 gene transfer may reduce these barriers to adenovirus-mediated myocardial gene transfer. PMID- 12542851 TI - Editorial. PMID- 12542850 TI - Lentiviral-mediated RNA interference. AB - RNA interference (RNAi) is a form of posttranscriptional gene silencing mediated by short double-stranded RNA, known as small interfering RNA (siRNA). These siRNAs are capable of binding to a specific mRNA sequence and causing its degradation. The recent demonstration of a plasmid vector that directs siRNA synthesis in mammalian cells prompted us to examine the ability of lentiviral vectors to encode siRNA as a means of providing long-term gene silencing in mammalian cells. The RNA-polymerase III dependent promoter (H1-RNA promoter) was inserted in the lentiviral genome to drive the expression of a small hairpin RNA (shRNA) against enhanced green fluorescent protein (EGFP). This construct successfully silenced EGFP expression in two stable cell lines expressing this protein, as analyzed by fluorescence microscopy, flow cytometry, and Western blotting. The silencing, which is dose dependent, occurs as early as 72 hr postinfection and persists for at least 25 days postinfection. The ability of lentiviruses encoding siRNA to silence genes specifically makes it possible to take full advantage of the possibilities offered by the lentiviral vector and provides a powerful tool for gene therapy and gene function studies. PMID- 12542852 TI - Antimicrobial and immunoregulatory functions of lactoferrin and its potential therapeutic application. AB - Lactoferrin is an iron-binding glycoprotein present in various secretions (eg. milk, tears, saliva,pancreatic juice, etc.). It is also stored in specific granules of polymorphonuclear granulocytes from which it is released following activation. Lactoferrin exerts a bactericidal activity by damaging the outermembrane of Gram-negative bacteria, as well as immunoregulatory functions by decreasing the release of interleukin-l (IL- 1), IL-2 and tumor necrosis factor alpha INF-alpha) and enhancing monocyte and natural killer cell cytotoxicity. Lactoferrin binds with high affinity to lipid A, the toxic moiety of the lipopolysaccharide, or endotoxin from Gram-negative bacteria Lipopolysacchride interaction with monocytes/ma phages results in the production and release of TNF alpha, that plays an important role in inducing septic shock In this respect, it has recently been demonstrated that lactoferrin inhibits the lipopolysaccharide interaction with CD14 on monocytes/macrophages by competition with the lipopolysaccharide binding protein. Therefore, besides its bactericidal activity, lactoferrin may also act by neutralizing the toxic effects of lipopolysaccharide and this protective role against endotoxin lethal shock has been demonstrated in animal models. Moreover, in vitro and in vivo neutralization of endotoxin by a human lactoferrin-derived peptide was also reported and lactoferrin or lactoferrin-derived peptides could represent useful tools for the treatment of endotoxin-induced septic hock. The recent production and characterization of monoclonal antibodies against different epitopes of human lactoferrin, including monoclonal antibodies selectively neutralizing lactoferrin binding to lipid A, may allow a better elucidation of the consequence of lactoferrin lipopolysaccharide interaction. PMID- 12542853 TI - Direct hemoperfusion with polymyxin B-immobilized fiber improves shock and hypoxemia during endotoxemia in anesthetized sheep. AB - This study evaluates the effect of direct hemoperfusion (DHP) using polymyxin B immobilized fibers (PMX-F) as an extracorporeal blood filter on systemic hypotension and lung injury during endotoxemia Sheep were anesthetized, intubated, mechanically ventilated with 50% oxygen and connected to the DHP system between the right femoral artery and left jugular vein. Group 1 (n = 6)sheep were infused with 10 microg/kg Escherichia coli endotoxin over a 30 min period. At the same time, sheep underwent DHP with PMX-F (Toraymyxin: PMX-20R) for 2 h at a flow rate of 60 ml/h. Group 2 (n = 6) sheep were infused with the same dose of endotoxin and treated with a sham column, in the same manner as those in group 1. DHP with PMX-F significantly improved and restored systemic pressure and arterial oxygen tension in group 1 sheep, although these values never returned to the baseline levels of group 2 sheep. Pulmonary hypertension and leukocytopenia were observed after endotoxin infusion in both groups, but there were no significant differences between these values. DHP with PMX-F significantly decreased the elevation of plasma nitric oxide products. The treatment with PMX-F improves shock and deteriorated oxygenation during endotoxemia, probably through the suppression of nitric oxide production. PMID- 12542854 TI - Sensitivity of sympathetically correlated spinal interneurons, renal sympathetic nerve activity, and arterial pressure to somatic and visceral stimuli after chronic spinal injury. AB - In the chronic stage of spinal cord injury in humans, both innocuous and noxious somatic and visceral stimuli can elicit severe autonomic dysreflexia characterized by potentially dangerous, sympathetically mediated, increases in arterial pressure. We hypothesized that a similar sympathetic hyperexcitability would be manifested in spinal sympathetic networks of chronically spinally transected rats. To test this hypothesis, we compared the responses of sympathetically correlated spinal interneurons and arterial pressure to both innocuous and noxious stimuli in acutely and chronically spinally transected rats. Experiments were conducted in anesthetized female rats, either within hours of T(3) spinal transection (rats with acute spinal transection) or one month after T(3) spinal transection (rats with chronic spinal transection). Sympathetically correlated spinal interneurons were identified by cross correlating their ongoing activity with simultaneously recorded renal sympathetic nerve activity. Cutaneous stimuli (either light brushing or noxious pinch) were delivered to a wide area of the ipsilateral side of the rat. Colorectal distension was used as a noxious visceral stimulus. The activity of sympathetically correlated interneurons was increased by stimulation of more of the body surface and decreased by stimulation of less of the body surface in rats with chronic spinal transection than in rats with acute spinal transection. Colorectal distension elicited greater increases in arterial pressure in chronically than acutely spinally-transected rats without exciting significantly more interneurons in those rats. These results suggest that spinal circuits undergo significant plastic changes in the chronic stage of spinal cord injury, and they provide a mechanism for the observation, in some human patients, that many stimuli, both noxious and non-noxious, applied caudal to the site of spinal injury increase sympathetic activity and arterial pressure. PMID- 12542855 TI - Neutralizing intraspinal nerve growth factor with a trkA-IgG fusion protein blocks the development of autonomic dysreflexia in a clip-compression model of spinal cord injury. AB - Increased intraspinal nerve growth factor (NGF) after spinal cord injury (SCI) is detrimental to the autonomic nervous system. Autonomic dysreflexia is a debilitating condition characterized by episodic hypertension, intense headache, and sweating. Experimentally, it is associated with aberrant primary afferent sprouting in the dorsal horn that is nerve growth factor (NGF)-dependent. Therapeutic strategies that neutralize NGF may ameliorate initial apoptotic cellular responses to the injury and aberrant afferent plasticity that occurs weeks after the injury. Subsequently, the development of autonomic disorders may be suppressed. We constructed a protein including the extracellular portion of trkA fused to the Fc portion of human IgG and expressed it using a baculovirus system. Binding of our trkA-IgG fusion protein was specific for NGF with a K(d) = 4.26 x 10(-11) M and blocked NGF-dependent neuritogenesis in PC-12 cells. We hypothesized that binding of NGF in the injured cord by our trkA-IgG fusion protein would diminish autonomic dysreflexia. Severe, high thoracic SCI was induced with clip compression and the rats were treated with intrathecal infusions (4 microg/day) of trkA-IgG or control IgG. At 14 days post-SCI, the magnitude of autonomic dysreflexia was assessed. Colon distension increased mean arterial pressure (MAP) in control rats by 46 +/- 2 from 96 +/- 5 mmHg. In contrast, MAP of rats treated with trkA-IgG increased by only 30 +/- 2 mmHg. Likewise, the MAP response to cutaneous stimulation was also reduced in rats treated with trkA-IgG (20 +/- 1 vs. 29 +/- 2). In contrast, trkA-IgG treatment had no effect on heart rate responses during colon distension or cutaneous stimulation. These results indicate that treatment with trkA-IgG to block NGF suppresses the development of autonomic dysreflexia after a clinically relevant spinal cord injury. PMID- 12542856 TI - Identification of regenerative tissue cables using in vivo MRI after spinal cord hemisection and schwann cell bridging transplantation. AB - The purpose of this study was to examine the feasibility of a non-invasive in vivo magnetic resonance imaging (MRI) procedure, performed at 1.5 T, to detect regenerative tissue cables in a rat spinal cord hemisection and Schwann cell (SC) bridging transplantation paradigm. Two months after implantation of a SC-seeded guidance channel (1.25 mm in diameter and 3.0 mm in length) into a T8 spinal cord hemisection-gap lesion, axial fast-spin echo (FSE) T2-weighted MR imaging (T2WI) was performed. Axial T2WI through the graft identified a circular area of low intensity surrounded by high-intensity signal within the guidance channel lumen. Correlative histological assessments of Toluidine blue-stained sections confirmed that the low-intensity signal represented a tissue cable, which, in most cases, contained a substantial number of myelinated axons oriented along the rostro caudal axis of the spinal cord. The percentage of guidance channel cross sectional area occupied by the tissue cable, expressed as the tissue cable index (TCI), was also determined from histological sections. Linear regression analysis of the TCI plotted relative to the number of myelinated axons revealed a strong positive correlation (r(2) = 0.85) between these two outcome measures. In addition, the sensitivity of MRI to detect regenerative tissue cables within guidance channels was 86%. These results demonstrate that (1). 1.5 T MR imaging performed 2 months after spinal cord hemisection and SC bridging transplantation is sensitive in detecting low-intensity regenerative tissue cables, and (2). the TCI strongly correlates with the extent of axonal regeneration into implanted SC seeded guidance channels. PMID- 12542857 TI - Changes in expression of amyloid precursor protein and interleukin-1beta after experimental traumatic brain injury in rats. AB - There is increasing evidence linking neurodegenerative mechanisms in Alzheimer's disease (AD) and traumatic brain injury (TBI), including increased production of amyloid precursor protein (APP), and amyloid-beta (Abeta) peptide. In vitro data indicate that expression of APP may be regulated in part by the inflammatory cytokine IL-1beta. To further investigate the mechanisms involved, we measured APP and IL-1beta protein levels and examined immunohistochemical localization of APP in brain tissue from rats subjected to controlled cortical impact (CCI) injury. Animals were examined at time intervals ranging from 3 h to 4 weeks after TBI. The 24-h time point revealed a dramatic increase in APP immunoreactivity, detected with both N- and C-terminal antibodies, in the hippocampus and cortex ipsilateral to injury. This finding was sustained up to 3 days post-injury. At these early time points, APP increase was particularly robust in the white matter axonal tracts. By 14 days after injury, APP immunoreactivity was not significantly different from sham controls in cortex, but remained slightly elevated in hippocampus. Western blot data corroborated early increases in hippocampal and cortical APP in injured versus control animals. Despite profound APP changes, no Abeta deposits were observed at any time after injury. Hippocampal and cortical IL-1beta increases were even more robust, with IL-1beta levels peaking by 6 h post-injury and returning to baseline by 24-72 h. Our results demonstrate that both APP and IL-1beta are rapidly elevated after injury. Because of the rapidity in the IL-1beta peak increase, it may serve a role in regulation of APP expression after TBI. PMID- 12542858 TI - Time-dependent changes in rat brain cholinergic receptor expression after experimental brain injury. AB - Alterations in neurotransmitter receptor expression in the central nervous system may contribute to physiological and behavioral deficits that follow traumatic brain injury (TBI). Previous studies from our laboratory have demonstrated significant and widespread deficits in alpha7* nicotinic cholinergic receptor (alpha7* nAChr) expression 2 days following cortical contusion brain injury. The purpose of this study was to evaluate changes in alpha7* nAChr expression over a wider range of post-TBI recovery intervals. Animals were anesthetized and subjected to a moderate cortical contusion brain injury (2 mm cortical compression). Animals were euthanatized at various post-TBI time intervals, ranging from 1 h to 21 days, and quantitative autoradiography was used to evaluate cholinergic receptor subtype expression in the cerebral cortex and hippocampus. As previously reported, the alpha7* nAChr was the most sensitive target of TBI-induced plasticity. Significant decreases in alpha-[(125)I] bungarotoxin (BTX) binding occurred as early as 1 h post-TBI, and persisted in some brain regions for up to 21 days. A kinetic analysis of changes in BTX binding, performed 2 days following brain injury, indicated that the binding deficits are not due to significant changes in receptor affinity. TBI-induced changes in alpha3*/alpha4* nACh receptors, muscarinic cholinergic receptors, and NMDA-type glutamate receptor expression were lower in magnitude, restricted to fewer brain regions and more transient in nature. Persistent deficits in alpha7* nAChr expression following TBI may contribute to impaired functional outcome following brain injury. PMID- 12542859 TI - Temporal and spatial profile of phosphorylated mitogen-activated protein kinase pathways after lateral fluid percussion injury in the cortex of the rat brain. AB - Mitogen-activated protein kinases (MAPK) play a crucial role in signal transduction that regulates gene expression through transcriptional factor activity. The purpose of this study was to investigate the temporal expression and topographic distribution of the activated MAPK pathways including extracellular signal-regulated protein kinase (ERK), c-Jun NH(2)-terminal kinase (JNK), and p38 MAPK following traumatic brain injury (TBI) in the cortex of the rat brain. Adult male Sprague-Dawley rats (300-400 g) were subjected to lateral fluid percussion injury of moderate severity (3.5-4.0 atm) using the Dragonfly device model (no. HPD-1700). Phosphorylated-MAPK protein levels were quantified using Western blot analysis. Topographic distribution of immunoreactivity for phosphorylated-MAPK was examined using immunohistochemistry. Our findings showed that TBI significantly increased the phosphorylated-ERK (p-ERK) and -JNK (p-JNK) levels, but not the -p38 (p-p38) protein levels in the cortex surrounding the injury site. The immunoreactivity for p-ERK and p-JNK immediately after TBI were localized in neurons. The immunoreactivity for p-JNK was uniformly but only transiently induced and returned to control levels 1 h after TBI. The immunoreactivity for p-ERK was confirmed up until 30 min after TBI in the superficial neuronal layers. Double immunostaining using a glial-specific marker demonstrated that p-ERK was prominent in astrocytes 6 h after TBI. The current results suggest that the ERK and JNK pathways, but not the p38 MAPK pathways are involved in signal transduction in the cortex following TBI. PMID- 12542860 TI - Alcohol consumption in traumatic brain injury: attenuation of TBI-induced hyperthermia and neurocognitive deficits. AB - Clinical and animal studies indicate that hyperthermia during or after traumatic brain injury (TBI) is associated with poor outcome. Alcohol intoxication, a complicating risk factor in many cases of head injury, has been found to both worsen or attenuate posttraumatic neural damage and outcome. The purpose of the present study was to determine whether chronic ethanol consumption would affect TBI-induced hyperthermia and deficits in spatial learning. TBI was produced by cortical contusion injury in adult male rats. We first characterized the TBI induced febrile response using probes implanted intraperitoneally (ip) or intracerebroventricularly for continuous biotelemetric recording of core body and brain temperatures and locomotor activity. In another experiment, rats, implanted with ip probes, were fed a liquid diet containing ethanol (5% w/v, 35% ethanol derived calories); control rats were pair-fed the isocaloric liquid diet (P-P). At 14 days after commencement of diet feeding, TBI or sham surgery was performed, and the ethanol-fed rats were divided into two groups: half were transferred to the isocaloric diet (E-P) and the other half remained on the ethanol-containing diet (E-E). TBI produced a significant febrile response in all rats, that persisted for at least 6 days in the E-P and P-P groups but lasted for only 2 days in the E-E group. When tested at 3-4 weeks after TBI, E-E rats required significantly fewer trials than E-P rats to reach criterion in the Morris water maze. In sum, continuous consumption of ethanol before and after TBI attenuated TBI-induced hyperthermia and deficits in spatial learning. Whereas the results suggest that this ethanol regimen may be neuroprotective, a causal relationship between the two outcomes remains to be determined. PMID- 12542861 TI - Intracerebral transplantation of marrow stromal cells cultured with neurotrophic factors promotes functional recovery in adult rats subjected to traumatic brain injury. AB - This study was designed to examine the effects of bone marrow stromal cells (MSCs) cultured in vitro with or without neurotrophic factors transplanted into adult male Wistar rats after traumatic brain injury (TBI). MSCs harvested from donor Wistar rats were cultured with either the culture medium containing brain derived neurotrophic factor (BDNF) and nerve growth factor (NGF) or the same culture media without these factors. Control and experimental animals were then traumatized by a controlled cortical impact. One day after the impact, either the placebo or the washed MSCs (1 x 10(6)) cultured with or without NGF and BDNF were transplanted adjacent to the site of injury. In addition, a nontreated group of rats was employed. Motor function of the animals was evaluated by the Rotarod test both before and after the injury. All animals were sacrificed 8 days after TBI, and the brain sections were stained by H&E as well as for immunohistochemistry. MSCs survived and migrated toward the injury site. The group treated with MSCs cultured with BDNF and NGF had a significantly higher number of engrafted cells than the group treated with MSCs cultured without BDNF and NGF (6.3 x 10(4) +/- 4250 compared to 4.1 x 10(4) +/- 3684; p < 0.05). In both groups, some transplanted MSCs showed positive staining for astrocytic (GFAP) and neuronal markers (Neu N and MAP-2). The groups treated with MSCs had better motor function than the groups receiving no treatment or receiving the placebo (PBS; p < 0.05); however, the improvement reached statistical significance only in the group treated with MSCs cultured with neurotrophic factors. These data suggest that more robust motor function described in rats subjected to TBI and treated with intracerebral transplantation of MSCs was achieved by the use of MSCs cultured with neurotrophic factors. PMID- 12542862 TI - NMDA receptor activation contributes to a portion of the decreased mitochondrial membrane potential and elevated intracellular free calcium in strain-injured neurons. AB - In our previous studies, we have shown that in vitro biaxial strain (stretch) injury of neurons in neuronal plus glial cultures increases intracellular free calcium ([Ca(2+)](i)) and decreases mitochondrial membrane potential (deltapsi(m)). The goal of this study was to determine whether strain injury, without the addition of exogenous agents, causes glutamate release, and whether NMDA receptor antagonists affect the post-strain injury rise in [Ca(2+)](i) and decrease in deltapsi(m). [Ca(2+)](i) and deltapsi(m) were measured using the fluorescent indicators fura-2 AM and rhodamine-1,2,3 (rh123). Strain injury of neuronal plus glial cultures caused an immediate 100-200 nM elevation in neuronal [Ca(2+)]i and a decline in neuronal deltapsi(m) by 15 min post-injury. Pretreatment with the NMDA receptor antagonist MK-801 (10 microM) attenuated the [Ca(2+)](i) elevation after mild, but not moderate and severe injury. MK-801 pretreatment reduced the decline in deltapsi(m) after mild and moderate, but not after severe injury. The NMDA receptor antagonist D-2-amino-5-phosphonopentanoic acid (APV; 100 microM) had effects similar to MK-801. Simultaneous measurement of [Ca(2+)](i) and deltapsi(m) demonstrated a significant correlation and a temporal relationship between [Ca(2+)](i) elevation and depression of deltapsi(m). We conclude that NMDA receptor stimulation contributes to some of the changes in [Ca(2+)](i) and deltapsi(m) after less severe strain injury. However, after more pronounced injury other mechanisms appear to be more involved. PMID- 12542863 TI - Pergamum: renowned medical center of antiquity. Origin of medical centers of today. PMID- 12542864 TI - Claudius Galenus of Pergamum: surgeon of gladiators. Father of experimental physiology. PMID- 12542865 TI - The effect of portal hypertension on the glycoprotein biosynthesis of rat gastric mucosa. AB - The aim of this study was to assess the influence of epidermal growth factor (EGF) on glycoprotein biosynthesis in portal hypertensive (PHT) gastric mucosa. Portal-vein ligation (PVL) for a period of 4 weeks was applied to 40 male Wistar rats to produce experimental portal hypertension. The rats were subdivided into four groups. Human EGF was administrated to these four groups of animals at a does of 0, 10, 25, and 50 microg/kg/day for 7 days. An additional group of 10 rats without PVL and EGF pretreatment was employed as a control. The severity of gross gastric mucosal lesions was evaluated macroscopically by a gross ulcer index. Glycoprotein biosynthesis of the gastric mucosa was determined by the incorporation rate of [(3)H]glucosamine. Quantitative changes of gastric mucosal hexosamines were also used for mucosal glycoproteins analyses. The gross mucosal damage was considerably greater in the PVL group without EGF pretreatment than in the EGF-pretreated groups (p <.05). The incorporation rate of [(3)H]glucosamine was significantly higher in the control group and the EGF-pretreated groups than in the PVL group without EGF pretreatment (p <.05). Moreover, the incorporation rate of [(3)H]glucosamine and the gastric mucosal hexosamine content were closely relevant to administration does of human EGF (p <.001). In addition, the reduction of glycoprotein biosynthesis was closely related to the increase in portal pressure (p =.001) and the severity of portal hypertensive gastropathy (p <.001). Our current study shows that the rate of incorporation of glucosamine is decreased in the PHT gastric mucosa and that EGF significantly stimulated glycoprotein synthesis in the PHT gastric mucosa. Accordingly, these findings may be helpful to explain the protective effect of EGF on the PHT gastric mucosa via increased glycoprotein biosynthesis in the stomach. PMID- 12542866 TI - Influence of mesh materials on collagen deposition in a rat model. AB - Alterations of the extracellular matrix (ECM) with its major component collagen are increasingly discussed as possible risk factors implicated in the development of abdominal-wall herniation. Because of the widespread use of alloplastic meshes for the surgical repair of hernias, an animal study was performed to analyze the influence of various mesh materials on the quantity and quality of collagen deposition. In 60 male Sprague-Dawley rats an abdominal replacement was performed using three different kinds of mesh materials: polyester (PE), a pure polypropylene (PP), and a composite mesh made of polypropylene and polyglactin (PG). A simple fascia suture repair served as control. The count of fibroblasts, the collagen/protein ratio, the type I/III collagen ratio, and the expression of basic fibroblast growth factor (b-FGF) at the interface were analyzed after 7, 21, and 90 days. The ratio of collagen to overall protein (microg/mg) showed significant differences comparing different mesh materials (sham controls 38.44 +/- 16.33 microg/mg, PE 68.5 +/- 23.8 microg/mg, PP 101.6 +/- 32.3 microg/mg, PG 49.6 +/- 11.6 microg/mg at day 90). The ratio of collagen type I/III increased over time in all groups. However, 90 days after mesh implantation the ratio was always significantly lowered compared to the controls. No significant difference was found comparing different mesh materials. The alteration of the scar composition is closely connected to an increased b-FGF expression. b-FGF and count of fibroblasts highly correlated (r =.95) and showed significant elevated levels compared to simple suture repair. The results of our study strongly support the notion that wound healing is affected by mesh implantation. The quality of the ECM deposition as determined by collagen type I/III ratio is impaired in general, whereas the quantity of ECM deposition is markedly influenced by the kind of mesh material. PMID- 12542867 TI - Chronic portal hypertension in the rat by triple-portal stenosing ligation. AB - A surgical technique based on the development of a triple stenosing ligation is used to worsen the complications inherent to the prehepatic chronic portal hypertension. The results have been compared with those obtained in rats with a single-portal stenosing ligation. An increase (p <.05) in the body, liver, spleen, and kidney weights as well as a decrease (p <.001) in the testes weight to body weight ratio were produced in both groups of animals. In addition, the variability in the obtained weights, particularly in the liver weight, stands out. The incidence of portosystemic and portohepatic collateral circulation and of the mesenteric venous vasculopathy increases in the animals with triple-portal stenosing ligation. The new proposed technique is a valid alternative to the classic one that used single portal stenosing ligation. PMID- 12542868 TI - Percutaneous central dual-lumen catheter for apheresis in the canine. AB - The dog model has been used extensively for preclinical hematopoietic stem-cell transplantation (HSCT) research. Apheresis of granulocyte colony-stimulating factor mobilized peripheral blood donor stem cells is increasingly being used for transplantation. We investigated the use of a percutaneous dual-lumen central venous catheter (DLC) for large-volume apheresis in the dog model. Fifteen dogs undergoing apheresis for HSCT had catheters placed percutaneously. Apheresis was performed in all 15 dogs without complications. In comparison to the previous techniques of arteriovenous shunts for apheresis, the DLC demonstrates comparable efficacy, is more efficient and cost-effective, and is associated with a lower complication rate. We demonstrate that the percutaneous DLC is the preferable method for apheresis in the dog model. PMID- 12542869 TI - Laparoscopic fluorometry: a new minimally invasive tool for investigation of the intestinal microcirculation. AB - The aim of this study was to develop and establish a new system of laparoscopic fluorometry for the purpose of investigating the intestinal microcirculation. In 25 pigs (German Landrace, 16-25 kg body weight), ischemia was established in two segments (A, irreversible; B, reversible ischemia; C, internal control) of the small intestine by a laparoscopic technique. Microcirculation in the segments was assessed by laparoscopy at a second-look operation 24 h later by means of the fluorescence system Endoscan. The fluorescence of the three bowel segments was measured by arbitrary dye fluorescence units (DFU) 15 min after starting reperfusion, before and after injection of sodium fluorescein (NaFlu, 0.25 mg/kg body weight). The dividing line between viable and nonviable bowel tissue was established from the inflow and outflow rates of NaFlu with the aid of ROC (receiver operating characteristic) curves. The specificity and sensitivity of the new method were evaluated by correlating the results with the viability of each intestinal segment as predicted by three laparoscopically experienced surgeons and by histological examination. By means of the calculated separation sharpness (fluorescence index at 2 min >0.5, outflow factor of NaFlu at 10 min >20%), the overall predictions of intestinal viability in all 25 animals achieved a sensitivity of 93.5% and a specificity of 94.1% by laparoscopic fluorometry, versus a sensitivity of 70.8% and a specificity of 87.5% for the prediction of bowel viability by ordinary laparoscopic technique. Used as an adjunct to conventional laparoscopy, laparoscopic fluorometry brought significant gains in sensitivity and specificity in the distinction between reversible and irreversible intestinal ischemia. PMID- 12542872 TI - Prefocal alignment improves stone comminution in shockwave lithotripsy. AB - BACKGROUND: The Dornier HM-3 machine continues to be one of the most effective lithotripters in use. However, tissue damage occurs in most, if not all, shockwave lithotripsy (SWL) treatments. Cavitation appears to contribute to desired stone comminution as well as to undesired tissue damage. Studies of cavitation in electrohydraulic shockwave lithotripters indicate that the greatest cavitation activity occurs, not at the geometric focus, F2, but at a site proximal to F2 by 1 to 3 cm. In clinical practice, however, stones are aligned with F2. MATERIALS AND METHODS: In vitro stone comminution, hemolysis, and free radical production were assessed along the focal axis, and pig kidneys treated with SWL in vivo were sectioned to determine the extent of hemorrhagic injury along the focal axis. Model gypsum stones received 200 shockwaves in vitro at 18 kV. RESULTS: At F2, the average number of fragments >1.5 mm was 1.3 +/- 0.5, and the weight loss was 11.3 +/- 1.1%. At 2 cm from F2 (F2-2 cm), these values increased to 4 +/- 2.8 and 16.1 +/- 4.2%, respectively. Samples of 10% hematocrit blood were similarly exposed. Hemolysis was equivalent at F2-2 cm (14.7 +/- 2.3%) and F2 (15.2 +/- 3%) but decreased significantly at all other positions. Samples of iodine solution received 1500 shockwaves at 20 kV. Hydroxyl radical production was greatest at F2-2 cm (0.384 +/- 0.035 microM) and decreased significantly distal to this position. The volume of tissue injury in pig kidneys was greatest with prefocal shockwave exposure. CONCLUSION: Stone comminution may be achieved more rapidly without greater tissue damage by a simple shift in stone alignment to F2-2 cm. PMID- 12542871 TI - A personal perspective on the origins of endourology and the endourological society. PMID- 12542873 TI - Shockwave lithotripsy for urinary stones in patients with urinary diversion after radical cystectomy. AB - BACKGROUND AND PURPOSE: During recent years, survival of patients with invasive bladder cancer has been improved by early diagnosis and radical treatment. Urinary lithiasis is not rare in patients who have been submitted to radical cystectomy and urinary diversion. We have demonstrated the effectiveness and safety of SWL for these patients. PATIENTS AND METHODS: We studied 11 patients who presented to our lithotripsy department suffering from urinary lithiasis after radical cystectomy. They were all cancer free at the time of treatment, and all underwent SWL on the Dornier HM-3 lithotripter as a first-line treatment. The mean stone burden was 1.85 cm(2), and the stone-to-patient ratio was 1:1. RESULTS: The stone-free rate 1 month after SWL was 63.7%. Patients who were not stone free underwent a second SWL, and the stone-free rate after the second SWL session was 81.8%. We performed percutaneous nephrolithotripsy in one patient after the second SWL session because of the large stone burden remaining (3.2 cm(2)). The remaining patient was submitted to ureterolithotomy. CONCLUSION: Application of SWL gives very good results in the treatment of urinary lithiasis in patients with a urinary diversion. Indeed, the results are equivalent to those achieved in patients without urinary diversion. PMID- 12542874 TI - Effect of single-dose subarachnoid spinal anesthesia on pain and recovery after unilateral percutaneous nephrolithotomy. AB - PURPOSE: We studied the impact of one dose of subarachnoid spinal analgesia on postoperative pain and recovery after percutaneous nephrolithotomy (PCNL). PATIENTS AND METHODS: Between August 1999 and May 2000, 20 PCNL patients were randomized into two groups: Group A (N = 9), who received preoperative subarachnoid spinal analgesia with morphine sulfate, and Group B (N = 11), who received no subarachnoid spinal analgesia. Both groups were given general anesthesia, and the nephrostomy site was infiltrated with bupivacaine hydrochloride. Stone size was similar in the two groups, as were ASA classification, age, and body mass index. Pain analog scales (maximum score 10) were completed preoperatively and on the day of surgery (D0), after 1 day (D1), and after 2 days (D2). The amount of morphine sulfate equivalents (MS eq) needed, the activity level, and adverse effects were recorded. RESULTS: In Group A, the average pain score on D0, D1, and D2 was 2.7, 3.7, and 1.4, respectively; in Group B, the average pain score was 4, 4.5, and 2, respectively (P > 0.05). The average MS eq used in Groups A and B were 8.3 v 33.8 (P = 0.002) on D0; 17.7 v 28.7 (P > 0.05) on D1; and 11.1 v 10.1 (P > 0.05) on D2. On D0, in Group A, 56% of the patients were ambulating and 11% complained of nausea, while in Group B, 0 were ambulating and 46% complained of nausea. CONCLUSIONS: A single preoperative dose of subarachnoid spinal analgesia provides a statistically significant decrease in postoperative parenteral pain medication and earlier ambulation. It also appears to reduce the amount of postoperative pain and decrease nausea. PMID- 12542875 TI - Radiation protection during percutaneous nephrolithotomy: a new urologic surgery radiation shield. AB - BACKGROUND AND PURPOSE: As endourology becomes an important part of the practice of urology, the use of fluoroscopic guidance has increased the exposure of urologists to the possibly deleterious effects of radiation. There is a need for a method of radiation protection for percutaneous nephrolithotomy (PCNL), as the exposure from radiation scatter may be significant, depending on the difficulty of establishing access. PATIENTS AND METHODS: We ascertained the effectiveness of a newly modified radiation shield during PCNL. Exposure readings were taken using a thermoluminescent dose monitor placed different distances from the radiation source during six PCNLs. We compared the exposure readings with and without the shield. RESULTS: The shield was able to reduce the radiation by an average of 96.1% at a distance of 25 cm and 71.2% at a distance of 50 cm from the source. CONCLUSION: The shield can be used as one step toward the goal of reducing surgeon radiation exposure. Other methods, such as dose-minimizing imaging protocols and adaptation of equipment optimized to reduce exposure, are also important measures in creating a safe environment for both the urologist and the patient. PMID- 12542876 TI - Tips and tricks for the management of retained ureteral stents. AB - BACKGROUND AND PURPOSE: Retained ureteral stents, especially those that are encrusted and associated with a stone burden, can be a difficult management problem. We review our experience and the different options employed for treating this complication. PATIENTS AND METHODS: From July 1998 to February 2002, 26 retained ureteral stents were managed in our department. The average patient age was 45.9 years (range 8-77 years). The average time the stent had been in place was 10.7 months (range 3-28 months). Prior to planning definitive therapy, a plain radiograph with tomographic views was reviewed. RESULTS: A guidewire or Glidewire was often placed adjacent to the stent in order to maintain ureteral access and in some cases was able to facilitate removal of the retained stent. The patients required an average of 2.7 endourologic procedures (range 1-4) performed at one or more sessions to remove the stent and all associated stone burden. If the stone burden could not be entirely removed then stent extraction and subsequent sessions were performed until stone-free status was achieved. Cystolitholapaxy was required to treat the distal component of stent encrustation in 20 cases. Percutaneous nephrolithotomy was performed in four patients, antegrade ureteroscopy with or without intracorporeal lithotripsy in four patients, retrograde ureteroscopy with or without laser lithotripsy in five patients, and extracorporeal shockwave lithotripsy in seven patients to treat the proximal component of stent encrustation. The stent could be removed in a single anesthetic session in 23 of 26 cases (88.5%). Analysis revealed that the major component of the encrustations was a combination of calcium oxalate and phosphate. CONCLUSION: Successful management of retained ureteral stents requires careful planning and may entail a combination of endourologic approaches. It is imperative to avoid using significant force, which can result in severe ureteral injury or breakage of the stent. If encrustations are present along the stent, we believe in treating the distal component prior to managing any proximal or ureteral components. PMID- 12542877 TI - Hand-assisted retroperitoneoscopic nephroureterectomy for upper urinary-tract urothelial tumors. AB - PURPOSE: We report our experience with hand-assisted retroperitoneoscopic nephroureterectomy for upper urinary-tract urothelial cancer. PATIENTS AND METHODS: Our initial 10 cases of clinical stage T(1)N(0)M(0) renal pelvic and ureteral tumors treated with hand-assisted retroperitoneoscopic nephroureterectomy are included in the present report. Nephrectomy was conducted retroperitoneoscopically with hand assistance via a lower-abdominal midline incision. Resection of the lower ureter together with the bladder cuff was performed as open surgery and the specimen was removed en bloc via the same incision. RESULTS: Hand-assisted retroperitoneoscopic nephroureterectomy was completed successfully in all 10 cases. The mean operating time was 456 +/- 90 minutes, and the mean estimated blood loss was 462 +/- 364 mL. The times to oral intake and walking were 1.5 +/- 0.5 days and 2.3 +/- 0.7 days, respectively. One case of renal vein injury, one case of pulmonary embolism, and three cases of wound infection were the complications. CONCLUSION: This is the first report of hand-assisted endoscopic nephroureterectomy using the retroperitoneal approach. The surgical technique seems quite reasonable because the lower-abdominal incision can be utilized, not only as a route for hand assistance, but also as a window for open surgery when resecting the distal ureter as well as for extracting the surgical specimen. PMID- 12542878 TI - Laparoscopic partial nephrectomy with a diode laser: porcine results. AB - PURPOSE: To develop a safe and effective technique for laparoscopic partial nephrectomy without need for hilar occlusion. MATERIALS AND METHODS: Laparoscopic transperitoneal lower-pole partial nephrectomy was performed in five 45- to 50-kg female farm pigs using a 980-nm diode laser. Standard transperitoneal access was obtained, and a four-port approach was used to perform a laparoscopic right partial nephrectomy using a diode laser (23 W) without hilar occlusion. The pigs were allowed to recover and 2 weeks later underwent a left laparoscopic partial nephrectomy. Postoperatively, renal function was monitored by serial serum creatinine measurements. Both kidneys and ureters were removed for ex-vivo retrograde pyelograms and histologic analysis. RESULTS: The 980-nm diode laser resulted in successful lower-pole partial nephrectomy without hilar occlusion in all 10 of the kidneys. In three cases, laser hemostasis was insufficient, and adjunctive hemostatic clips were necessary to stop bleeding. The mean operative time was 126 minutes, and the mean laser time was 84 minutes. An average of 23% (range 13%-33%) of the kidney parenchyma was resected. The mean blood loss was 150 mL (range 50-300 mL). There was no evidence of urinary extravasation on ex vivo retrograde pyelograms at 2 weeks in any of the kidneys. CONCLUSION: Laparoscopic partial nephrectomy without hilar occlusion using the 980-nm diode laser is feasible in the porcine model. Because adjunctive hemostatic measures may be necessary in some cases, clinical trials in humans should be limited to small exophytic tumors. PMID- 12542879 TI - Role of early endourologic management of tuberculous ureteral strictures. AB - BACKGROUND AND PURPOSE: Tuberculous ureteral stricture causing progressive obstructive uropathy commonly complicates renal tuberculosis. The aim of our study was to evaluate the effectiveness of early ureteral stenting or percutaneous nephrostomy (PCN) in patients with tuberculous ureteral strictures. PATIENTS AND METHODS: Seventy-seven patients (84 renal units) with tuberculous ureteral strictures were analyzed. We evaluated the final outcome of involved kidneys with three different managements: medication only (N = 37), medication plus ureteral stenting (N = 28), or medication plus PCN (N = 19). RESULTS: The overall nephrectomy rate was 51%. In patients treated with medication only, the nephrectomy rate was 73%, whereas the nephrectomy rate was 34% in patients treated with medication plus early ureteral stenting or PCN. The rate of reconstructive surgery for ureteral strictures also was significantly different for patients treated with medication only (8%) and those receiving medication plus early ureteral stenting or PCN (49%). Spontaneous resolution of the strictures was noted in 6 of the 12 renal units that were managed with early ureteral stenting. CONCLUSIONS: Early ureteral stenting or PCN in patients with tuberculous ureteral strictures may increase the opportunity for later reconstructive surgery and decrease the likelihood of renal loss. PMID- 12542880 TI - A bioabsorbable self-expandable, self-reinforced poly-L-lactic acid urethral stent for recurrent urethral strictures: long-term results. AB - BACKGROUND AND PURPOSE: Biodegradable urethral stents have been used clinically for some years mainly in order to prevent postoperative urinary retention after minimally invasive thermotreatment of benign prostatic hyperplasia. We previously reported a 15-month-follow-up pilot study of a bioabsorbable self-expandable, self-reinforced poly-L-lactic acid (SR-PLLA) urethral stent in combination with optical urethrotomy in the treatment of recurrent urethral strictures. The present aim was to evaluate the long-term results of this new treatment modality. PATIENTS AND METHODS: A series of 22 patients with severe urethral strictures (mean number of earlier urethrotomies 2.5) were involved. An SR-PLLA spiral stent with a bioabsorption time of 10 to 12 months was inserted into the urethra at the stricture site immediately after optical urethrotomy. Uroflowmetry, measurement of postvoiding residual urine, urine culture, and urethroscopy were performed preoperatively and at 1 and 3 months and then every 3 months up to 12 months, with subsequent follow-up visits every 6 months. The mean follow-up was 46 months. RESULTS: The treatment was successful in 8 of the 22 patients (36%). Six recurrences were found within the stented area and 15 outside. Patients with failure have since been treated with several urethrotomies and repeat self dilatations or free skin urethroplasties. CONCLUSIONS: The results of the use of a bioabsorbable SR-PLLA urethral stent for the treatment of recurrent strictures were encouraging. Without this additional therapy, the recurrence rate of strictures might have been much higher. The main problem was sudden collapse of the stent, possibly induced by outer compression. A new generation of bioabsorbable stents is already under development. PMID- 12542881 TI - How did the endoscopic video camera change our practice in transurethral resection of the prostate? A retrospective study of 200 cases. AB - PURPOSE: Video camera-assisted transurethral resection of the prostate (TURP) is an invaluable teaching tool and provides physical comfort to the surgeon. However, whether it improves technical skills and reduces complications rates and hospital stay has not been well documented. PATIENTS AND METHODS: We retrospectively reviewed 100 cases of video camera-assisted (VCA) TURP and 100 cases of non-video camera-assisted (NVCA) TURP randomly chosen from 1490 cases performed over a 5-year period. The patients' mean age was similar in the two groups (70 years in the NVCA group and 71 years in the VCA group). RESULTS: The mean weight of prostatic chips was 33.2 g in the VCA and 19.02 g in the NVCA group (P < 0.05). The mean operative time was identical in the two groups (VCA 41 minutes and NVCA 42.2 minutes). The mean postoperative hospital stay was shorter in the VCA group than in the NVCA group (1.56 and 3.39 days, respectively) (P < 0.05), as was the urinary drainage time (1.68 and 4.04 days, respectively) (P < 0.05). In the NVCA group, three patients had profuse hemorrhage necessitating blood transfusions and two had TURP syndrome, while in the VCA group, one patient had TURP syndrome and no patient experienced intraoperative or postoperative hemorrhage. Urethral stricture was less frequent in the VCA group than in the NVCA group (1% and 3%, respectively). CONCLUSIONS: With video camera-assisted TURP, we resected larger prostate glands with minimal intraoperative bleeding, better hemostasis, and a shorter hospital stay. PMID- 12542882 TI - Vapor resection: a good alternative to standard loop resection in the management of prostates >40 cc. AB - BACKGROUND AND PURPOSES: The morbidity of transurethral vesection of the prostate (TURP) necessitates constant attempts at modifications to the standard equipment and technique. Patients with larger prostates (>40 cc) need a longer time for the procedure, and the blood loss, requirement for irrigation fluid, and incidence of postoperative complications tend to be greater. We report on the safety and efficacy of TURP with the thick vapor resection loop compared with the standard wire loop in comparable groups of patients with prostates >40 cc. PATIENTS AND METHODS: We randomized 100 patients with benign prostatic hyperplasia (BPH) into two groups of 50 each which were similar in age. The inclusion criteria included an indication for prostatectomy and prostate size >40 cc. Patients who were on finasteride preoperatively (six) and those who had histopathologic adenocarcinoma of the prostate (three) were excluded. Preoperative evaluation included assessment of International Prostate Symptom Score (IPSS), prostate volume by abdominal ultrasonography, maximum flow rate (Q(max)), and residual urine volume. The two groups were similar in the signs and symptoms of BPH. Patients in Group 1 underwent transurethral vapor resection of the prostate (TUVRP) using the vapor resection loop (Wing trade mark; Richard Wolf, Germany), while patients in Group 2 underwent TURP using a standard wire loop. All procedures were performed by consultant urologists with equivalent experience. We used a Martin ME 401 (Gebruder Martin, Tuttlingen, Germany) electrosurgical generator with settings of 120 to 150 Watts and 50 to 70 W for cutting and coagulating, respectively, for the thick loop and 70 to 80 W and 40 to 50 W for the standard loop. Operating time, resected tissue weight, duration of catheterization, nursing contact time, hospital stay, hemoglobin change, serum sodium concentrations, and any complications were noted and analyzed using the Kruskal-Wallis paired variables test, and P values were calculated. P value <0.04 was considered significant. The IPSS, Q(max), and residual urine volume were reevaluated at 6 months and 1 year after the procedure and compared for the two groups. RESULTS: The median prostate volume was 63 cc and 54 cc in Groups 1 and 2, respectively, and the median resected weight was 20 and 19 g, respectively (P = NS). The differences in operating time (median 45 v 60 minutes; P < 0.0001), intraoperative irrigant use (15 v 21 L; P < 0.0001), and intraoperative blood loss (median 52.5 v 150 mL; P < 0.0001) in the two groups were statistically significant. Peroperative blood loss was estimated by the indicator dilution method of Freedman et al in three 5-mL samples of the irrigant fluid and arriving at the average of the three values and adjusted for the volume of irrigant fluid used. The differences in postoperative irrigant (P < 0.01) and catheter duration (P = 0.04) were also significant. Two patients received blood transfusion in Group 2 and none in Group 1. The difference in the change in hemoglobin and serum sodium postoperatively in the two groups was not statistically significant. The complications were dysuria in six patients lasting for a month and one capsular perforation in Group and incontinence lasting for 6 months in two patients and stricture in one patient in Group 2. The efficacy, assessed with IPSS, Q(max), and residual urine volume, was comparable at 6 months and at 1 year. CONCLUSION: The use of a thick vapor resection loop for TURP coupled with higher generator settings as recommended for them, especially for prostates >40 cc, is beneficial, as it significantly reduces operating time, blood loss, irrigant requirement, nursing contact time, and duration of catheterization, besides providing clear vision during surgery and ease of resection. There was no reduction in efficacy or increase in complications. PMID- 12542883 TI - The graying of... PMID- 12542884 TI - Urban seventh graders and smoking: a health risk behavior assessment. AB - Health risk behaviors undertaken in adolescence, such as smoking, can have a lasting impact on both short-term and long-term health developments. To better describe the health risk behaviors of an adolescent urban population, a study was conducted at two parochial middle schools in the southwest section of Philadelphia. The study purpose was to describe (1) the types of health risk behaviors being undertaken by a seventh grade student population, (2) the frequency of health risk behaviors, and (3) the age of initiation of the health risk behavior, e.g., age when they first began smoking. A descriptive, correlational study was undertaken with 106 urban seventh graders (ages of 11-13 years) from two seventh grade classes. Using the Youth Risk Behaviors Surveillance System Questionnaire, information was collected about health risk behaviors. Correlational statistics indicated significant correlations between students who smoke and alcohol use, dieting, and not feeling safe in their neighborhood. Interestingly, girls were more likely to use smoking as their primary method of weight control and dieting. Because the long-term impact of smoking is known to be negative, nurses should provide support to this at-risk population and also should be prepared to educate adolescents who come under their care. PMID- 12542885 TI - Birth weight, neonatal morbidities, and school age outcomes in full-term and preterm infants. AB - In this prospective longitudinal study, birth weight and neonatal morbidities were evaluated relative to a broad range of school age outcomes. Fully 188 infants, 151 who were preterm, were recruited at birth, stratified by birth weight and socioeconomic status, and were followed until age 8 with a 97% retention rate. A gradient relationship was found among birth groups, with full term children earning the highest scores. The very low birth weight and extremely low birth weight groups were equivocal in all scores except visual perception. The findings also were consistent with a pattern of nonverbal learning disability (Rourke, 1995) in which there is evidence of math underachievement and adequate performance in verbal, reading, and spelling scores. Children who had bronchopulmonary dysplasia, chronic lung disease, intraventricular hemorrhage, and sepsis differed from children without these neonatal morbidities, with an average of 10-20 points below the mean. PMID- 12542886 TI - An evaluation of college and low-income youth writing together: self-discovery and cultural connection. AB - Although the health and healing effects of writing have been documented in the literature, most of the studies have focused primarily on individuals writing alone. This formative evaluation is a component of an intervention reported elsewhere. The evaluation describes the experience of low-income youth and college students (n = 7) writing in a group during a 10-week workshop. The results revealed the development of protective processes of self-esteem, self efficacy, coping strategies, social support, and cultural connections. In the weekly 2-hr writing sessions, using the Amherst Writers and Artists method, participants were encouraged to write their stories in their own voices in response to prompts, followed by reading aloud and positive feedback. At the end of the 10 weeks participants were interviewed about their experience within the group and outside the group. Analysis of interviews revealed two themes that emerged from the experience of writing together: connection to self through feelings, reflection, and behaviors; and connection to others through learning and empathy. The results suggest that writing in a group using a specific approach facilitated emotional catharsis, increased self knowledge, coping strategies, and understanding and appreciating of others. PMID- 12542887 TI - Liposomally targeted cytotoxic drugs for the treatment of cancer. AB - Phospholipid spherules composed of lipid bilayer membranes entrapping a central aqueous core were first described more than 30 years ago (Bangham et al 1965). The term liposome was coined in 1968 (Sessa & Weissmann 1968) and the first suggestions that these vesicles might have potential as vehicles for targeted drug delivery for a range of diseases, including cancer, appeared shortly afterwards (Gregoriades et al 1974; Gregoriades 1976a, b). However, the process of turning this expectation into a clinical reality has suffered a number of setbacks and has taken more than a quarter of a century. In the process, new types of liposomes with favourable in-vivo pharmacokinetics and biodistribution patterns have been generated (Lasic & Papahadjopoulos 1995). Many of these preparations have been subjected to extensive examination and an increasing number of agents have entered clinical trials. In this review, we will trace the development of those liposomes that are currently undergoing (or are about to undergo) clinical evaluation. PMID- 12542888 TI - Alpha-monoisostearyl glyceryl ether enhances percutaneous penetration of indometacin in-vivo. AB - Molecules that reversibly remove the barrier resistance of skin enhance penetration. alpha-Monoisostearyl glyceryl ether (GE-IS) is a novel compound that can be used as a non-ionic surfactant and increases percutaneous penetration of indometacin in rat abdominal skin in-vitro. The present study investigated GE-IS induced enhancement of indometacin penetration in-vivo. When 1% GE-IS in propylene glycol was applied to rat abdominal skin, serum and muscle concentrations of indometacin increased markedly. Anti-inflammatory activities of test solutions containing both indometacin and GE-IS were investigated in experimental models of acute and chronic inflammation. Application of indometacin with GE-IS to the skin produced greater inhibitory effects on carrageenan-induced rat paw oedema, UV-induced erythema in guinea-pigs, and adjuvant arthritis in rats, compared with application of indometacin alone. The results suggest that GE IS enhances penetration in-vivo and improves the anti-inflammatory effects of indometacin in animal models. Thus, GE-IS might contribute to the development of cosmetic or medical formulations to improve transfer of bioactive substances to hypodermal sites. PMID- 12542889 TI - Topical delivery of plasmid DNA using biphasic lipid vesicles (Biphasix). AB - The development of non-invasive methods for the delivery of vaccines through the skin will greatly improve the safety and the administration of human and veterinary vaccines. In this study we examined the efficiency of topical delivery of plasmids by assessing the localization of gene expression using luciferase as a reporter gene and induction of immune responses using a plasmid encoding for the bovine herpesvirus type-1 glycoprotein D (pgD). Topical administration of plasmids in a lipid-based delivery system (biphasic lipid vesicles--Biphasix) resulted in gene expression in the lymph node, whereas with intradermal injection, antigen expression was found in the skin. Following administration of plasmid with the gene gun, antigen expression was observed in both the skin as well as in the draining lymph nodes. Transcutaneous immunization with pgD formulated in biphasic lipid vesicles elicited gD-specific antibody responses and a Th2-type cellular response. In contrast, immunization by the intradermal route resulted in the stimulation of a Th1-type response. These findings have implications for both vaccine design and tailoring of specific immune responses. PMID- 12542890 TI - Biorelevant dissolution testing of St John's wort products. AB - In contrast to chemically defined drugs, most herbal medicinal products (HMPs) are poorly characterized in terms of their pharmaceutical properties. In many cases it is assumed that the plant extract as a whole is the active moiety, since it is often difficult to identify the individual components responsible for the pharmacological activity and even more difficult to assess synergies among the various components. However, where the active components have been identified, it should be possible to compare products with respect not only to content uniformity but also to their biopharmaceutical properties. The aim of this study was to investigate and compare the dissolution characteristics of several St John's wort products under biorelevant conditions. Components of St John's wort known, or suspected, to play a role in its antidepressant activity include phloroglucines, naphthodianthrones and the flavonoids. Since these groups have a broad spectrum of polarity and solubility, dissolution was studied for representative compounds from each group. Although the labelling indicates that several of the products studied should be pharmaceutically equivalent, dissolution under biorelevant conditions revealed that they have quite different release profiles and cannot be considered switchable. It was concluded that biorelevant dissolution testing can be a powerful tool for comparing HMPs as well as synthetically produced drug products. PMID- 12542891 TI - Metabolism and excretion of centbutindole (neuroleptic) in rats after oral administration. AB - The metabolism and excretion of centbutindole was studied in Sprague-Dawley rats after oral administration. The percentage of dose excreted was monitored over 4 days. Efforts were also directed towards evaluating the stability of centbutindole in various in-vitro biomatrices. Centbutindole was found to be metabolised in rats and only a negligible amount of parent drug was excreted. The percentage of the dose excreted in bile and faeces was 0.2% and 0.6%, respectively. No drug was found in urine after oral administration, but after an intravenous dose only 0.0012% of the dose was eliminated through urine. Two metabolites, dealkylated metabolite and hydroxy metabolite, were identified by ion spray LC/MS/MS, using a combination of parent ion and product ion scanning techniques. The major routes of metabolism of centbutindole include reduction of the carbonyl functional group in the butyrophenone side chain and N-dealkylation of the butyrophenone side chain attached to the pyrazinopyridoindole ring nitrogen at position 2. The hydroxy metabolite was excreted negligibly in bile, although it was present in the form of glucuronide conjugates more in comparison to its free form. PMID- 12542892 TI - Transbilayer transport of a propyltrimethylammonium derivative of diphenylhexatriene (TMAP-DPH) in bovine blood platelets and adrenal chromaffin cells. AB - The membrane fluorescent probe N-((4-(6-phenyl-1,3,5 hexatrienyl)phenyl)propyl)trimethylammonium (TMAP-DPH) has an additional three carbon spacer between the fluorophore and the trimethylammonium substituent of 1 (4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH). As a basic study to clarify the transport mechanism of amphiphilic quaternary ammoniums, we observed the characteristics of the transbilayer transport of TMAP-DPH in bovine blood platelets and bovine adrenal chromaffin cells using the albumin extraction method. We compared these inward transport rates with those of TMA-DPH. TMAP-DPH crossed into the cytoplasmic layers of the membranes more slowly than TMA-DPH after rapid binding to the outer halves of the plasma membranes. The transport rate markedly depended on temperature. Time to reach the half-maximal incorporated amount of TMAP-DPH increased threefold accompanied by an increase in the concentration from 0.2 to 1.5 microM. The transport was stimulated significantly by various types of membrane perturbations such as modification of sulfhydryl-groups by N-ethylmaleimide and benzyl alcohol-induced increase in the fluidity of the lipid bilayer. The saturation phenomenon suggested the presence of the regulatory process in the transbilayer transport of TMAP-DPH. PMID- 12542893 TI - Tocolytic activity of formoterol against premature delivery in mice. AB - The tocolytic activity of formoterol (eformoterol), a long-acting potent beta(2) adrenoceptor agonist, was assessed in pregnant mice, with determination of uterine effects on the 15th and 16th days of gestation. For examination in the lipopolysaccharide-induced premature delivery model, osmotic pumps filled with formoterol or saline solution were implanted subcutaneously under the back skin. The mice were sacrificed 18-20 h thereafter, and the numbers of fetuses in the uteri and the newborn were counted. The uteri, amniotic membranes and placenta were also rapidly removed for determination of IL-6 concentrations. Furthermore, the effect of formoterol on IL-6 secretion from mouse amnion cells was determined. Formoterol and ritodrine inhibited contraction responses of isolated mouse uteri and their intravenous administration resulted in lowered uterine motility. Lipopolysaccharide (30 microg mL(-1)/mouse) induced premature delivery, attributable to increased IL-6 secretion, and formoterol suppressed this. Doses of 5-500 microg/mouse thus reduced the number of prematurely delivered newborn, and 50 microg/mouse also depressed IL-6 secretion. On histopathologic analysis, the marked oedema and slight haemorrhage in the mouse cervix induced by lipopolysaccharide were reduced by administration of the beta(2)-adrenoceptor agonist. Neither formoterol (10(-7)-10(-5) M) nor ritodrine (10(-7)-10(-5) M) influenced spontaneous secretion of IL-6 in amnion cells. However, at 10(-7) and 10(-5) M, and 10(-6) and 10(-5) M, respectively, they inhibited lipopolysaccharide-induced IL-6 secretion and this inhibitory effect was competitively reversed by addition of ICI-118,551 (beta(2)-adrenoceptor antagonist), but not atenolol (beta(1)-adrenoceptor antagonist). These findings strongly suggest that formoterol can suppress premature delivery mediated by its actions on IL-6 secretion. PMID- 12542894 TI - Inhibition of urinary bladder motility by a spinal action of U-50488H in rats. AB - We examined the effect of a kappa agonist, U-50488H, upon the bladder motility of anaesthetized rats. The frequency of distension-induced rhythmic bladder contractions was reduced by the intravenous (10 mg kg(-1)) or intrathecal (10-100 microg) administration of U-50488H. The effect of intravenous U-50488H was inhibited by pre-treatment with nor-binaltorphimine (10 mg kg(-1), s.c.). The inhibition of bladder contractions by intrathecal U-50488H (30 microg) was eliminated with the concomitant use of nor-binaltorphimine (10 mg kg(-1), s.c.), and diminished by reserpine (4 mg kg(-1), i.p.), yohimbine (10 microg, i.t.) or methysergide (20 microg, i.t.). The amplitude of bladder contractions induced by an electrical stimulation of the pontine micturition centre was not inhibited by intrathecal U-50488H (30 and 100 microg). These results suggested that a kappa agonist could inhibit micturition reflex as well as other opioids, and at least part of the inhibition was due to the diminishment of bladder sensation based on the activation of the descending monoaminergic systems through the spinal kappa opioid receptors. PMID- 12542895 TI - Autoantibodies to malondialdehyde-modified low-density lipoprotein in patients with angiographically confirmed coronary artery disease. AB - Oxidised low-density lipoprotein (LDL) has physicochemical properties that are pivotal in atherosclerotic plaque formation. As a consequence, antioxidant regimens may prove an important therapy in the prevention and treatment of cardiovascular disease. Since oxidised LDL is immunogenic, the aims of our study were to measure serum IgG titres to malondialdehyde-modified LDL (MDA-LDL) in patients with coronary artery disease (CAD) and control subjects and assess their potential as a clinical marker for coronary atherosclerosis and, consequently, antioxidant intervention. Serum IgG titres to MDA-LDL were measured in patients with angiographically confirmed CAD (n = 40) and aged-matched controls (n = 40) by enzyme-linked immunosorbant assay (ELISA). Titres were calculated and expressed as both the difference and the ratio of blanked absorbance units (AU) for IgG binding to MDA-LDL and native LDL. For the control population, median IgG titres were 0.26 AU (interquartile range (IQR) 0.20-0.46 AU) and 5.34 (IQR 3.40 8.58), respectively, while the patient population had median IgG titres of 0.30 AU (IQR 0.20-0.47 AU) and 5.08 (IQR 3.30-9.66), respectively. Both sets of calculated titre values were not significantly different between the two groups (P = 0.60 and 0.82, respectively). In conclusion, this study could not establish any significant association between circulating autoantibody titres to MDA-LDL and coronary atherosclerosis. Therefore, the diagnostic value of autoantibodies to oxidised LDL remains unclear. PMID- 12542896 TI - The effect of the isomers of cyclo(Trp-Pro) on heart and ion-channel activity. AB - Cyclo(L-Trp-L-Pro) has shown potential for use in the treatment of cardiovascular dysfunction. The aim of the study was to determine the effects of the isomers of cyclo(Trp-Pro) - cyclo(L-Trp-L-Pro), cyclo(L-Trp-D-Pro), cyclo(D-Trp-L-Pro) and cyclo(D-Trp-D-Pro) - on heart and ion-channel activity. The effects on L-type Ca(2+)-channel, Na(+)-channel and inward rectifier K(+)-channel activity were determined by using the whole-cell patch-clamp technique on myocytes of guinea pig origin. Dependence on the membrane potential in terms of Ca(2+)-channel activity was also investigated. A modified Langendorff method was used to determine the effects of the isomers on heart rate, coronary flow, duration of ventricular tachycardia and arrhythmia, time to sinus rhythm and QRS interval on the rat isolated heart. Cyclo(L-Trp-L-Pro), cyclo(L-Trp-D-Pro) and cyclo(D-Trp-D Pro), 100 microM, showed agonism towards Ca(2+)-channel activity, while cyclo(D Trp-L-Pro) caused a blockage of the current. The action of cyclo(D-Trp-L-Pro) was shown to be independent of membrane potential. No significant effect (P > 0.05) on the inward rectifier K(+) current was observed in the presence of cyclo(L-Trp D-Pro) and cyclo(D-Trp-D-Pro), while antagonism was noted in the presence of cyclo(L-Trp-L-Pro) and cyclo(D-Trp-L-Pro). All isomers showed antagonist effects on the Na(+) channel. No adverse effects were noted on chronotropic effects in the presence of 200 microM cyclo(L-Trp-L-Pro) and cyclo(D-Trp-D-Pro) (P > 0.05), while cyclo(L-Trp-D-Pro) significantly increased the heart rate. Cyclo(D-Trp-L Pro) significantly reduced the heart rate (P < 0.05). In addition, no significant effects were observed on the coronary flow rate in the presence of the isomers. All isomers significantly reduced the duration of ventricular tachycardia and arrhythmia, as well as the time to sinus rhythm. Furthermore, no change in the QRS intervals was noted in the presence of the isomers in comparison with the control, with a significant increase being noted for cyclo(D-Trp-D-Pro) (P < 0.05) in reference to the other isomers. The isomers thus show antiarrhythmic potential and may manifest as novel agents in the treatment of cardiovascular dysfunction, since a decrease in ventricular fibrillation may reduce the mortality rates in acute myocardial infarction. PMID- 12542897 TI - Rapid inhibition of the contraction of rat tail artery by progesterone is mediated by inhibition of calcium currents. AB - Progesterone induced rapid relaxation of KCl-contracted tail artery helical strips from rats. The effect was dose dependent, with an IC50 (inhibitory concentration which produces 50% of the maximal response) of 8.9 microM progesterone. The actions of progesterone were not blocked by bicuculline, indicating that in this tissue the non-genomic actions of progesterone were not mediated via a gamma-aminobutyric acid (GABA)-A receptor. Fura-2 was used to measure intracellular calcium levels ([Ca(2+)](i)) in isolated vascular smooth muscle cells (VSMC). Incubation of cultured VSMC for 15 min with progesterone (10 microM) resulted in an inhibition of the KCl-induced [Ca(2+)](i )increase. The whole-cell patch-clamp technique was used to examine Ca(2+)-channel currents in the membrane of isolated VSMC. Progesterone suppressed the L-type Ca(2+)-channel currents in cells held at a potential of -40 mV. The effects of progesterone were quickly reversed by washout in all three experimental protocols suggesting that these effects on vascular tissues are non-genomic. The correlation of the effects on all these preparations, their time course and reversibility suggested that the rapid relaxation of the rat tail artery induced by progesterone is mediated at least in part by inhibition of L-type calcium channels, leading to inhibition of calcium responses in the VSMC of this tissue. PMID- 12542898 TI - Aspirin and sodium salicylate inhibit proliferation and induce apoptosis in rheumatoid synovial cells. AB - Aspirin has been reported to induce apoptosis in a variety of cell lines. In this study, we examined whether aspirin and sodium salicylate inhibit cell growth and induce apoptosis in rheumatoid synovial cells. Synovial cells were obtained from patients with rheumatoid arthritis, and the cells were treated with aspirin or sodium salicylate (0.1-10 mM) for 24 h. Cell proliferation and viability were assessed by 5-bromo-2'-deoxyuridine incorporation and by 4-[3-(4-iodophenyl)-2-(4 nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate (WST-1) assay, respectively. The apoptosis of synovial cells was identified by DNA fragmentation assay and terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) assay. Aspirin and sodium salicylate suppressed the proliferation (IC50 (concentration causing 50% inhibition of cell proliferation): 2.1 and 1.2 mM, respectively) and reduced the viability (IC50: 2.0 and 1.4 mM, respectively) of synovial cells in a concentration-dependent manner at 0.3-10 mM. Furthermore, they induced DNA fragmentation and increased the number of TUNEL-positive synovial cells. These results suggest that aspirin and sodium salicylate can inhibit the proliferation of rheumatoid synovial cells through induction of apoptosis. PMID- 12542899 TI - Cimetidine: antioxidant and metal-binding properties. AB - Cimetidine is one of the most potent H(2) receptor antagonists for inhibiting excessive histamine-induced acid secretion and is currently used worldwide to treat peptic ulcers. In this study, levels of free radicals were assessed and the ability of cimetidine to act as an antioxidant was determined using nitroblue tetrazolium assay and lipid peroxidation assays. Free radical generation in the brain is promoted by the presence of iron, as occurs in the Fenton reaction. The results show that cimetidine reduces the generation of superoxide anion formed in the nitroblue-tetrazolium assay. In addition, cimetidine (1 mM) is able to reduce the iron-induced rise in lipid peroxidation in rat brain homogenates. Electrochemistry, UV/Vis spectroscopy and HPLC experiments show metal-ligand interactions between cimetidine and transition metals. The results imply that cimetidine provides a neuroprotective effect by binding to iron and copper, thus making them unavailable for free radical production. PMID- 12542900 TI - Effects of acute renal failure induced by uranyl nitrate on the pharmacokinetics of intravenous theophylline in rats: the role of CYP2E1 induction in 1,3 dimethyluric acid formation. AB - In rats with acute renal failure induced by uranyl nitrate, the hepatic microsomal cytochrome P450 (CYP) 2E1 and CYP3A23 increased 2-4- and 4-times, respectively, CYP2C11 decreased to 80% of control, but the levels of CYP1A2 and CYP2B1/2 were not changed. It has been reported that theophylline was metabolized to 1,3-dimethyluric acid by CYP1A2 and CYP2E1 and 1-methylxanthine via CYP1A2, which was metabolized further to 1-methyluric acid via xanthine oxidase in rats. Hence, it was expected that the formation of 1,3-dimethyluric acid would show an increase in rats with renal failure as a result of induction of CYP2E1. The pharmacokinetics of theophylline were compared in control rats and rats with renal failure after intravenous administration of aminophylline, 5 mg kg(-1) as theophylline. In rats with renal failure, the plasma concentrations of theophylline were considerably lower and the resultant total area under the plasma concentration-time curve from time zero to time infinity (AUC(0- infinity )) of theophylline was significantly smaller (2,200 vs 1,550 microg min mL(-1)) compared with control rats. In rats with renal failure, the plasma concentrations of 1,3-dimethyluric acid were considerably higher and the resultant AUC(0-6 h) of 1,3-dimethyluric acid was significantly greater (44.4 vs 456 microg min mL(-1)) compared with control rats. Moreover, the AUC(0-6 h, 1,3-dimethyluric acid)/AUC(0 infinity, theophylline) ratio increased from 2.02% in control rats to 29.4% in rats with renal failure. The in-vitro intrinsic 1,3-dimethyluric acid formation clearance was significantly faster in rats with renal failure (734 vs 529 10(-6) mL min(-1)) compared with control rats using hepatic microsomal fraction. The results led us to conclude that in rats with uranyl nitrate-induced renal failure after the administration of aminophylline, 5 mg kg(-1) as theophylline, there was an increase in the formation of 1,3-dimethyluric acid as a result of an increase in CYP2E1 expression. PMID- 12542901 TI - Continuous infusion versus intermittent administration of cefepime in patients with Gram-negative bacilli bacteraemia. AB - The objective of this study was to compare the pharmacokinetics of cefepime administered by continuous infusion and intermittent injection regimens. A prospective, randomized, cross-over study of ten patients with Gram-negative bacilli bacteraemia was conducted. All patients were randomized to receive cefepime either as a 4-g continuous infusion over 24 h for 48 h or a 2-g bolus administered intermittently intravenously every 12 h for 48 h. After 48 h the patients received the alternative dose regimen. Cefepime pharmacokinetic studies were carried out during hours 36-48 after the start of both regimens. All of the pathogens isolated from the blood in 7 patients had a minimum inhibitory concentration (MIC) < 1 microg mL(-1). In both regimens, the serum cefepime concentrations at all time points were higher than the MIC for the pathogens isolated from this study. For the continuous infusion arm, the highest steady state concentration was 49.80+/-18.40 microg mL(-1) and the lowest steady-state concentration was 41.42+/-16.48 microg mL(-1). The steady-state concentrations were greater than 4 times the MIC of 8 microg mL(-1). For the intermittent injection regimen, the mean trough concentration was 4.74+/-3.99 microg mL(-1). The mean serum cefepime concentration was above 8 microg mL(-1) for 81.66% of the dosing interval. Therefore, we conclude that either continuous infusion or intermittent injection can be used as an effective mode of cefepime administration to achieve bactericidal activity. PMID- 12542902 TI - Synthesis and radical scavenging of novel magnolol derivatives. AB - We have investigated the developdment of potential antioxidants based on magnolol, a naturally occurring biphenolic obtained from the bark of Magnolia officinalis. A series of aminomethylated derivatives of magnolol were synthesized under the aromatic Mannich reaction. In-vitro testing for diphenyl-p picrylhydrazyl (DPPH) scavenging and chemiluminescence assays in whole cell models revealed that the pyrrolidyl-containing magnolols (2b (5,5'-diallyl-3 (pyrrolidin-1-ylmethyl)-biphenyl-2,2'-diol), 3a (5,5'-diallyl-3,3'-bis (pyrrolidin-1-ylmethyl)-biphenyl-2,2'-diol) and 4c (5,5'-diallyl-3-(morphorin-4 ylmethyl)-3'-(pyrrolidin-1-ylmethyl)-biphenyl-2,2'-diol)) displayed promising free radical scavenging effects as compared with magnolol. The results from compound 4c indicated that the naturally occurring component was suitable to be a lead compound toward promising antioxidants. PMID- 12542905 TI - Some practical aspects of providing a diagnostic service for respiratory chain defects. AB - The oxidative phosphorylation system (OXPHOS) is organized into five multi protein complexes, comprising four complexes (I-IV) of the respiratory chain and ATP synthase (complex V). OXPHOS has a vital role in cellular energy metabolism and ATP production. Enzyme analysis of individual OXPHOS complexes in a skeletal muscle biopsy remains the mainstay of the diagnostic process for patients suspected of mitochondrial cytopathy. Practical guidelines are presented to provide optimal conditions for performance of laboratory investigations and a reliable diagnosis. A fresh muscle biopsy is preferable to a frozen muscle sample because the overall capacity of the OXPHOS system can be measured in a fresh biopsy. In about 25% of patients referred for muscle biopsy to our centre, reduced substrate oxidation rates and ATP+creatine phosphate production rates were found without any defect in complexes I-V and the pyruvate dehydrogenase complex. Investigation of frozen muscle biopsy alone may lead to false-negative diagnoses in many patients. In some patients, it is necessary to investigate fibroblasts for prospective diagnostic purposes. An exact diagnosis of respiratory chain defects is a prerequisite for rational therapy and genetic counselling. Provided guidelines for specimen collection are followed, there are now reliable methods for identifying respiratory chain defects. PMID- 12542906 TI - Proteomics and neuromuscular diseases: theoretical concept and first results. AB - Although many patients suspected of suffering disturbances of the mitochondrial energy metabolism have been investigated, only a fraction of these patients have been diagnosed at the molecular level. Introduction of new techniques like proteomics will be necessary to understand the various clinical and biochemical aberrations in the field of mitochondrial disorders. Two-dimensional electrophoresis is the first, important step in the proteomics strategy. Separation of soluble proteins is performed on the basis of isoelectric point (net charge) in one direction and on molecular mass in the other. The technique provides an overview of the majority of proteins expressed in a sample (e.g. muscle biopsy, muscle cell or mitochondrial fraction). Once an abnormal spot is observed in the gel the responsible protein can be identified by analysing a limited part of its amino acid sequence by mass spectrometry. We optimized two dimensional (2D) gel electrophoresis to obtain high resolution 2D-maps and tested the reproducibility of the technique. Potentially, this new technique is capable of identifying novel mitochondrial diseases and defining their molecular basis. PMID- 12542907 TI - Proton nuclear magnetic resonance spectroscopy of body fluids in the field of inborn errors of metabolism. AB - Proton nuclear magnetic resonance (NMR) spectroscopy of body fluids has been successfully applied to the field of inborn errors of metabolism. This technique has the advantage of minimal sample pretreatment not requiring extraction or derivatization steps. Moreover, the spectrum provides a comprehensive metabolic profile of proton-containing, low-molecular-weight metabolites. The sensitivity limit is in the low micromolar range. This allows diagnosis of many inborn errors of metabolism. This review explains the key features of the NMR spectrum and reviews the available literature on metabolic diseases. Three novel diseases have been delineated with the technique. Relevant parts of the spectra from the urine samples of patients with these diseases are shown. NMR spectroscopy may develop to become a key tool in a metabonomics approach in clinical biochemistry. PMID- 12542908 TI - Brain-specific proteins in cerebrospinal fluid for the diagnosis of neurodegenerative diseases. AB - Neurodegenerative disorders have traditionally been classified according to clinical criteria, e.g. as dementia syndromes (the best known is Alzheimer's disease) or as movement disorders (e.g. Parkinson's disease). Another subdivision is based on recent insights into the respective pathogenetic mechanisms, leading to the recognition of so-called tauopathies and alpha-synucleinopathies. It is this increased knowledge of the underlying (neuro)pathological mechanisms that has sparked interest in studies aimed at the identification of disease-specific biomarkers in cerebrospinal fluid (CSF) for this field of neurological disorders. This review deals with the recent progress that has been made in identification, quantification and subsequent validation of brain-specific proteins in CSF for the diagnosis of various neurodegenerative disorders. Development of disease specific CSF biomarkers will undoubtedly add to the process of differential diagnosis early in the course of the disease. PMID- 12542909 TI - Pharmacogenetic and clinical aspects of dihydropyrimidine dehydrogenase deficiency. AB - Dihydropyrimidine dehydrogenase (DPD) is the initial and rate-limiting enzyme in the catabolism of 5-fluorouracil (5FU). A deficiency of DPD is increasingly being recognized as the cause of an important pharmacogenetic syndrome. The importance of DPD deficiency in the aetiology of unexpected severe 5FU toxicity has been demonstrated by the fact that, in 39-59% of cases, decreased DPD activity could be detected in peripheral blood mononuclear (PBM) cells. It was observed that 55% of the patients with a decreased DPD activity suffered from grade IV neutropenia compared with 13% of the patients with a normal DPD activity (P = 0.01). Furthermore, toxicity developed significantly earlier in patients with low DPD activity than in patients with normal DPD activity (10.0 +/- 7.6 versus 19.1 +/- 15.3 days, P < 0.05). In patients suffering from severe 5FU-associated toxicity, 11 mutations have been identified in DPYD, including one splice-site mutation (IVS14 + 1G-->A), one nonsense mutation (E386X), four missense mutations (M166V, V335L, I560S, D949V) and five polymorphisms (C29R, R21Q, S534N, I543V, V732I). Considering the common use of 5FU in the treatment of cancer patients, the severe 5FU-related toxicities in patients with a low DPD activity and the high prevalence of the IVS14 + 1G-->A mutation, analysis of the DPD activity in PBM cells or screening for the IVS14 + 1G-->A mutation should be routinely carried out prior to the start of treatment with 5FU. PMID- 12542910 TI - Genetics of hyperhomocysteinaemia in cardiovascular disease. AB - Homocysteine, a sulphur amino acid, is a branch-point intermediate of methionine metabolism. It can be degraded in the transsulphuration pathway to cystathionine, or remethylated to methionine via the remethylation pathway. In both pathways, major genetic defects that cause enzyme deficiencies are associated with very high plasma homocysteine concentrations and excretion of homocystine into the urine. Mildly elevated plasma homocysteine concentrations are thought to be an independent and graded risk factor for both arterial occlusive disease and venous thrombosis. Genetic defects in genes encoding enzymes involved in homocysteine metabolism, or depletion of important cofactors or (co)substrates for those enzymes, including folate, vitamin B(12) and vitamin B(6), may result in elevated plasma homocysteine concentrations. Plasma homocysteine concentrations are also influenced by dietary and lifestyle factors. In the last decade, several studies have been conducted to elucidate the genetic determinants of hyperhomocysteinaemia in patients with cardiovascular disease. We report on both environmental and genetic determinants of hyperhomocysteinaemia and give a detailed overview of all the genetic determinants that have been reported to date. PMID- 12542911 TI - Is serum cystatin C the marker of choice to predict glomerular filtration rate in paediatric patients? AB - BACKGROUND: It has been suggested that serum cystatin C (cyst-C) concentration provides a better indication of changes in glomerular filtration rate (GFR) than does serum creatinine concentration. METHODS: Because of conflicting results as to the usefulness of cyst-C, we compared the GFRs calculated from serum cyst-C, inulin clearance and endogenous creatinine clearance in children. GFRs calculated from cystatin concentration, inulin clearance following a single injection and endogenous creatinine clearance using Jaffe and enzymic methods were compared in 66 children (1.3-21.9 years) with a variety of renal disorders. Receiver operating curve analysis was used to determine the cut-off value that would give the best discrimination between normal and decreased GFR. RESULTS: The serum cyst C concentration ranged from 0.66 to 7.61 mg/L (median 1.94). Serum creatinine Jaffe concentration (creat-J) ranged from 38 to 871 micro mol/L (median 105) and creatinine enzymatic concentration (creat-E) ranged from 28 to 862 micro mol/L (median 126). The linear correlation coefficient (R) of 1/cyst-C versus GFR (R = 0.937) did not differ from either that of 1/creat-J versus GFR (R = 0.918) or that of 1/creat-E versus GFR (R = 0.901). These coefficients had overlapping confidence intervals. The areas under the curve for cyst-C, creat-J and creat-E were 0.967, 0.977 and 0.924, respectively, and were not significantly different from each other. For cyst-C, the optimal cut-off was 1.1 mg/L. CONCLUSIONS: Serum cyst-C is equivalent to creat-J and creat-E as a marker for estimating the GFR in the paediatric population studied. PMID- 12542912 TI - Cobalamin-binding proteins in normal and cobalamin-deficient older subjects. AB - BACKGROUND: The causes of cobalamin (vitamin B(12)) deficiency in older people are only partly understood. We investigated the role of the cobalamin-binding proteins and tested the hypothesis that low saturated transcobalamin concentration is an early marker of cobalamin deficiency. METHODS: We measured saturated (holo) and unsaturated (apo) transcobalamin and haptocorrin concentrations in healthy middle-aged volunteers, healthy older volunteers, cobalamin-deficient older volunteers and cobalamin-deficient older patients. RESULTS: Holo and apo concentrations of transcobalamin and haptocorrin were similar in healthy middle-aged and older subjects. Holotranscobalamin concentrations were significantly decreased in cobalamin-deficient subjects but did not differ between healthy volunteers and patients. Furthermore, the relative amount of cobalamin on transcobalamin (i.e. holotranscobalamin/holotranscobalamin + holohaptocorrin) was similar in all four groups. CONCLUSIONS: Abnormalities of the cobalamin-binding proteins are not a cause of cobalamin deficiency in the aged. Plasma holotranscobalamin concentration did not differ between stages of cobalamin deficiency in older persons. Therefore, plasma holotranscobalamin is not an early marker of cobalamin deficiency in older people and has no additional value in the diagnostic work-up of reduced plasma cobalamin concentrations in older people. PMID- 12542913 TI - Combination therapy in childhood leukaemia: in vitro studies of thiopurines and inhibitors of purine metabolism on apoptosis. AB - BACKGROUND: Methotrexate (MTX) followed by 6-mercaptopurine (6MP) is one of the best known combinations for the treatment of childhood acute lymphoblastic leukaemia. Tiazofurin (TF) and 6-thioguanine (TG) are also used as chemotherapy agents in the treatment of malignancies. We have examined the induction of apoptosis by combinations of these drugs to gain more insights into their efficacy in the treatment of malignancies. METHODS: The induction of apoptosis was examined in Molt-4, a human malignant acute lymphoblastic T-cell line. The cells were exposed to increasing drug concentrations at various exposure times. Annexin V/FITC and propidium iodide (PI) were used as markers for apoptosis and cell death. Annexin V/FITC positive and PI positive cells were detected by flow cytometric analysis. RESULTS: Sequential 24-h exposure with MTX (0.005-0.02 micro mol) followed by 6MP (1-10 micro mol) and 24-h exposure with TF (5-20 micro mol) followed by TG (0.5-2 micro mol) showed a more than additive induction of apoptosis compared with single-drug exposure. Simultaneous administration of the drugs does not show an additive effect on apoptosis. CONCLUSIONS: The results of this study indicate that sequential administration of MTX before 6MP and of TF before TG may be essential for therapeutic success in the treatment of leukaemia. PMID- 12542914 TI - An improved procedure for testing for assay linearity. AB - BACKGROUND: Statistical testing of assay linearity using replicate measurements made on binary mixtures of samples provides no information about the significance of any detected deviations from linearity, which may often be desirable. METHOD: Performance of additional statistical analysis of the assay data. RESULT: Objective measures for non-linearity and the minimum detectable deviation from linearity of the assay are obtained. CONCLUSION: A few additional calculations from the available data make it possible to assess whether or not an assay method needs improvement to achieve adequate performance within its intended application. PMID- 12542915 TI - Kinetics of myo-inositol loading in women of reproductive age. AB - BACKGROUND: Myo-inositol plays a key role in an important intracellular signalling pathway. A deranged myo-inositol metabolism has been associated with neural tube defects. A myo-inositol loading test was performed to investigate the kinetics in healthy women of reproductive age. METHODS: Five healthy non-obese females [mean age (standard deviation: SD) 22.8 (2.2) years] were recruited at the University Medical Center Nijmegen. Blood samples were drawn fasting and at 20, 40, 60, 90, 180 and 270 min after ingestion of 100 mg/kg body weight of myo inositol. Urine samples were collected before myo-inositol loading and at 180 and 270 min post-loading. Samples were analysed for serum myo-, epi- and scyllo inositol and glucose concentrations by gas chromatography. Plasma insulin concentrations were determined by radio-immunoassay. Random intercept models were fitted to evaluate the data. RESULTS: The estimated myo-inositol and scyllo inositol concentrations both reached maximum values at 180 min post-loading, respectively: mean (SD) 101.5 (9.2) micro mol/L and 1.09 (0.11) micro mol/L. The estimated plasma insulin and serum glucose concentrations decreased slightly but significantly during the experiment: P < 0.0001 and P < 0.05, respectively. At 180 and 270 min post-loading, urinary myo-inositol concentrations were increased and urinary glucose concentrations were unchanged. CONCLUSIONS: Myo-inositol enters the bloodstream quickly after oral ingestion and a small amount of myo inositol is converted to scyllo-inositol. The synthesis of glucose from myo inositol could not be detected by serum measurements. These data can be used in further research into the association between myo-inositol and neural tube defects. PMID- 12542916 TI - Measurement of thiopurine S-methyltransferase activity in human blood samples based on high-performance liquid chromatography: reference values in erythrocytes from children. AB - BACKGROUND: Monitoring 6-thiopurine S-methyltransferase (TPMT; EC 2.1.1.67) activity is especially important when patients are treated with 6-thiopurine drugs, since severe bone marrow toxicity may be induced if patients have deficient TPMT activity. METHODS: We have developed a method based on high performance liquid chromatography (HPLC) for the measurement of TPMT activity in various cell types: erythrocytes (RBC), human peripheral blood mononuclear cells (pMNC) and human malignant lymphoblasts (Molt-F4). The enzymatic activity is measured by the amount of 6-methylmercaptopurine formed, using 6-mercaptopurine (6MP) as substrate and S-adenosylmethionine as co-substrate. RESULTS: The K(m) values calculated for 6MP were 0.54 (RBC), 0.85 (pMNC) and 0.65 (Molt-F4 cells) mmol/L. The K(m) values for S-adenosylmethionine were 11.9 (RBC), 16.4 (pMNC) and 6.65 (Molt-F4 cells) micro mol/L. The assay variation was 8.2-17%. TPMT activity was determined in a control group of 103 children and young adults (44 female, 59 male). The values observed were (mean +/- standard deviation): female children and young adults, 15.1 +/- 4.8 pmol/10(7) cells per h (n = 44); male children and young adults, 15.8 +/- 6.4 pmol/10(7) cells per h (n = 59). No gender or age differences were found. CONCLUSION: The HPLC-based method enables the rapid screening of TPMT activities in large groups of patients treated with 6 thiopurines. PMID- 12542917 TI - Quantitative detection of peripheral thyroglobulin mRNA has limited clinical value in the follow-up of thyroid cancer patients. AB - BACKGROUND: As recurrences after treatment for differentiated thyroid cancer can occur many years after thyroidectomy, periodic monitoring of serum thyroglobulin (Tg) levels is performed in these patients. However, autoantibodies that can interfere with Tg immunoassays occur in the blood of approximately 25% of these patients. Several earlier reports suggest that measuring Tg mRNA by reverse transcriptase polymerase chain reaction (RT-PCR) could be of value, especially in patients with Tg autoantibodies. METHODS: Using an earlier described, real-time quantitative Taqman RT-PCR assay, Tg mRNA concentrations were assessed in peripheral blood taken from 58 patients treated for thyroid cancer and from two healthy controls. RESULTS: In all tested samples Tg mRNA could be found. No correlation between serum Tg protein and Tg mRNA could be found. Tg mRNA concentrations did not differ between serum Tg-negative and Tg-positive patients. No differences in the number of patients with high or low Tg/beta-actin ratios were found between the groups of patients without, (131)I uptake on whole-body scan, or patients with thyroid bed uptake, uptake elsewhere in the neck, or distant metastases with or without regional uptake (P = 0.871). CONCLUSIONS: We were not able to confirm earlier positive reports on the clinical value of Tg mRNA measurement for the monitoring of patients treated for thyroid cancer. PMID- 12542918 TI - Stability studies of ubiquinol in plasma. AB - BACKGROUND: Ubiquinol is a sensitive redox marker in the first line of the antioxidative defence mechanism and is increasingly being measured in oxidation studies. Because of its apparent instability during storage and processing, we compared various storage conditions. METHOD: Blood was collected from three volunteers into tubes containing EDTA; it was then separated at 4 degrees C and cryopreserved with saccharose (final concentration 6 g/L). Aliquots were stored with or without glutathione or butylated hydroxytoluene at -20 degrees C and -80 degrees C. RESULTS AND CONCLUSION: Ubiquinol in samples stored at -20 degrees C was not stable; however, it was stable when stored at -80 degrees C, even without addition of antioxidant. By contrast, alpha-tocopherol was stable under all conditions studied. PMID- 12542919 TI - Moderate citrullinaemia without hyperammonaemia in a child with mutated and deficient argininosuccinate synthetase. AB - In a patient with microcephaly, feeding problems and restlessness, moderately increased serum and urine citrulline concentrations were observed. Protein and allopurinol loading did not result in additional indications for a urea cycle defect. The diagnosis of citrullinaemia was made at both the enzyme and DNA level, resulting from a novel mutation in the argininosuccinate synthetase gene. The fact that the patient has not suffered from severe deterioration, and that there were only minor abnormalities in metabolite concentrations, suggests that the argininosuccinate synthetase capacity was less affected in vivo than in vitro. In vitro nuclear magnetic resonance investigation suggested an active acetylation mechanism for citrulline. This case illustrates the importance of performing extensive biochemical and molecular investigations in order to reach a definitive diagnosis, particularly in instances of moderate citrullinaemia. PMID- 12542920 TI - Reversal of clinical symptoms and radiographic abnormalities with protein restriction and ascorbic acid in alkaptonuria. AB - There is no definitive treatment protocol for alkaptonuria. A patient with alkaptonuria was treated with ascorbic acid (0.5 g/day) from the age of 4 years. He developed episodes of severe recurrent joint pain at 9.5 years of age after which a protein-restricted diet (1.3 g/kg/day) was started. Protein restriction in combination with ascorbic acid therapy (1 g/day in two divided doses) resulted in a significant decrease but not a normalization of the urinary homogentisic acid excretion. Joint pain resolved and the radiological evidence of 'moth-eaten' irregularities on the articular surface in both knees disappeared. He is currently well, growing normally and in nitrogen balance. Our findings document a reversal of bone abnormalities and clinical symptoms in a case of alkaptonuria. The results should be confirmed in a larger study. We suggest that protein restriction should be applied in combination with ascorbic acid in affected patients as soon as joint pain occurs. PMID- 12542921 TI - Are we draining the life from our patients? PMID- 12542922 TI - The Surgical Infection Society guidelines on antimicrobial therapy for intra abdominal infections: an executive summary. AB - The Surgical Infection Society last published guidelines on antimicrobial therapy for intra-abdominal infections in 1992 (Bohnen JMA, et al., Arch Surg 1992;127:83 89). Since then, an appreciable body of literature has been published on this subject. Therefore, the Therapeutics Agents Committee of the Society undertook an effort to update the previous guidelines, primarily using data published over the past decade. An additional goal of the Committee was to characterize its recommendations according to contemporary principles of evidence-based medicine. To develop these guidelines, the Committee carried out a systematic search for all English language articles published between 1990 and 2000 related to antimicrobial therapy for intra-abdominal infections. This literature was reviewed individually and collectively by the Committee, and categorized according to the type of study and its quality. Additional articles published prior to 1990 were also utilized when necessary. By a process of iterative consensus, the Committee developed provisional guidelines for antimicrobial therapy for intra-abdominal infections based on this evidence. Following extensive review by members of the Society, these guidelines were approved for publication in final form by the Council of the Surgical Infection Society. This executive summary delineates the Society's current recommendations for antimicrobial therapy of patients with intra-abdominal infections. Topics discussed include the selection of patients needing therapeutic antimicrobials, duration of antimicrobial therapy, acceptable antimicrobial regimens, and identification and treatment of higher-risk patients. Guidelines for patient selection and specific antimicrobial regimens were based on relatively good evidence, but those regarding optimal duration of therapy and treatment of higher risk patients relied mostly on expert opinion, since there was a paucity of high quality studies on those issues. Relevant areas for future investigation include the safety, convenience, and cost-effectiveness of available antimicrobial regimens for lower-risk patients, and better means for identifying and treating higher-risk patients with intra-abdominal infections. PMID- 12542923 TI - The Surgical Infection Society guidelines on antimicrobial therapy for intra abdominal infections: evidence for the recommendations. AB - Revised guidelines for the use of antimicrobial therapy in patients with intra abdominal infections were recently developed by the Therapeutic Agents Committee of the Surgical Infection Society (Mazuski et al., Surg Infect 2002;3:161-173). These were based, insofar as possible, on evidence published over the past decade. The objective of this document is to describe the process by which the Committee identified and reviewed the published literature utilized to develop the recommendations and to summarize the results of those reviews. English language articles published between 1990 and 2000 related to antimicrobial therapy for intra-abdominal infections were identified by a systematic MEDLINE search and an examination of references included in recent review articles. If current literature with regard to a specific issue was lacking, relevant articles published prior to 1990 were identified. All prospective randomized controlled trials, as well as other articles selected by the Committee, were evaluated individually and collectively. Data with regard to patient numbers, types of infections, and results of interventions were abstracted. Studies were categorized according to their design, and all included trials were graded according to quality. On the basis of this evidence, the Committee formulated recommendations for antimicrobial therapy for intra-abdominal infections and graded those recommendations. After receiving comments from invited reviewers and the general membership of the Society, the guidelines were finalized and submitted to the Council of the Surgical Infection Society for approval. The final recommendations related to the selection of patients needing therapeutic antimicrobials, acceptable antimicrobial regimens, duration of antimicrobial use, and the identification and treatment of higher-risk patients. Although numerous publications pertaining to these topics were identified, but nearly all of the prospective randomized controlled trials represented comparisons of different antimicrobial regimens for the treatment of intra-abdominal infections. A few prospective trials evaluated the need for therapeutic antimicrobial therapy in patients with peritoneal contamination following abdominal trauma. The quality of these prospective trials was highly variable. Many did not limit enrollment to patients with complicated intra-abdominal infections, lacked blinding of treatment assignment, did not provide a complete description of the criteria used to determine therapeutic success or failure, failed to identify the reasons why patients were excluded from analysis, or did not include an intention-to-treat analysis. For many issues, no prospective randomized controlled trials were encountered, and guidelines had to be formulated using evidence from studies with historical controls or uncontrolled data, or on the basis of expert opinion PMID- 12542924 TI - Influence of antiseptic agents on interleukin-8 release and transmigration of polymorphonuclear neutrophils in a human in vitro model of peritonitis. AB - BACKGROUND: The aim of this study was to evaluate the influence of taurolidine (TAU) and polyhexanid (POLY) on basic inflammatory reactions during peritonitis by using an in vitro model of human peritoneum. MATERIALS AND METHODS: Human umbilical vein endothelial cells (HUVEC) and human peritoneal mesothelial cells (HPMC; concentration: 2x10(5)/cm2) were brought on a collagen-coated filter insert with 3-microm pore size (HUVEC on the bottom, HPMC on the top), thus resulting in a two-chamber peritoneal model. After 5 days, confluence of the cells was reached, and HPMC were stimulated with 0.5 mL of TNF-alpha (10 microg/mL) for 4 h. Afterwards, 0.5 mL of TAU (1% and 2%) or 0.5 mL of POLY (0.1% and 0.2%) solution were added to the upper (HPMC) compartment. Polymorphonuclear neutrophils (PMN, 10(6)/mL) were placed in the lower compartment 1 h later. After 2 and 6 h, aliquots were taken from the upper compartment and transmigrated PMN were counted. Interleukin-8 (IL-8) concentrations were measured in both compartments by chemiluminescent enzyme immunometric assay. Expression of the adhesion molecules P-selectin and intercellular adhesion molecule-1 (ICAM-1) was assessed by immunohistochemistry. Controls were either TNF-alpha-stimulated HPMC without any antiseptic agents, or stimulated HPMC where TNF-alpha had been substituted by culture medium. Each experiment was performed in triplicate. RESULTS: Stimulation with TNF-alpha led to a time-dependent increase of IL-8 secretion to the apical compartment resulting in a gradient between both chambers, as well as to a time-dependent increase of PMN transmigration and expression of adhesion molecules. IL-8 gradients and PMN migration were significantly higher as compared to the other groups (p<0.05). After substitution of the stimulus by culture medium, significantly less IL-8 was measured in both compartments. PMN transmigration was almost absent (p<0.05). Addition of POLY and TAU led to comparable low IL-8 gradients with concomitant low PMN transmigration. The initially detected expression of adhesion molecules significantly decreased during the observation time. The IL-8 gradient in all groups correlated significantly with PMN transmigration (r=0.74226; p<0.0001). CONCLUSION: The diminished IL-8 response together with low PMN transmigration rates after addition of TAU and POLY may reflect either antiinflammatory effects or cellular damage. PMID- 12542925 TI - High-pressure gradients generated by closed-suction surgical drainage systems. AB - BACKGROUND: Closed suction drains remain in widespread use in surgical practice. There have been reports of complications associated with their use. We sought to characterize the pressure-generating capacities of three commonly used closed suction drainage systems. MATERIALS AND METHODS: Three commonly used closed suction surgical drainage systems were studied: Davol Reliavac 400 Evacuator, Jackson-Pratt Closed Wound Suction Drainage System, and Snyder Hemovac 400. Each drainage system was connected to a pneumatic pressure transducer, and pressure measurements were made. Measurements were made with the drain reservoirs at varying degrees of fullness. Measurements were also made while "stripping" the drains. RESULTS: In all three systems, maximal negative pressures (-71 to -175 mm Hg) were generated with the reservoirs empty of fluid. Pressure generation by all drains decreased as the volume of fluid in the reservoir increased. In all cases, drain "stripping" was associated with a transient elevation in drain pressure (p<0.05). In two out of three drains, stripping led to a significant residual increase in static drain pressure. CONCLUSION: Closed suction drains are capable of generating high pressures that may contribute to some complications associated with their use. Closed suction drainage systems differ with regard to their generation of negative pressure. PMID- 12542926 TI - In vitro assessment of microbial barrier properties of Dermabond topical skin adhesive. AB - BACKGROUND: Several different techniques are used for wound closure. Cyanoacrylate tissue adhesives are less invasive and easy to apply, and the cosmetic results are comparable to or better than the conventional methods for wound repair. The purpose of this study is to demonstrate that Dermabond Topical Skin Adhesive, 2-octylcyanoacrylate (a registered trademark of Ethicon) is an effective barrier against the penetration of microorganisms in an in vitro model. MATERIALS AND METHODS: Octylcyanoacrylate adhesive was evaluated in vitro as a barrier to microbial penetration using a strike through test. Agar media containing pH sensitive dye was used. The dye changed color in the presence of acidic microbial metabolic products. Octylcyanoacrylate adhesive was applied to the agar surface to form uniformly thick films. A total of 300 single layered films and an additional 300 triple layered films were evaluated. A 10-microL aliquot of inoculum containing at least 1x10(3) cfu was applied to the surface of each film. Plates were incubated at 37 degrees C for 72 h and were observed for growth and color change every 24 h during the incubation period. RESULTS: A series of binomial calculations were performed, varying the level of effectiveness of the test and the level of statistical confidence. Of the 600 test articles evaluated, 598 retained their patency at the end of 72 h. One individual test film was invalidated due to extrinsic contamination. The data presented indicated that octylcyanoacrylate tissue adhesive provided a barrier to microbial penetration with 95% confidence of 99% efficacy for 72 h in this in vitro model. CONCLUSION: The results of these in vitro experiments supported the hypothesis that octylcyanoacrylate tissue adhesive is an effective barrier to microbial penetration by gram-positive and gram-negative motile and nonmotile species. PMID- 12542927 TI - Economic consequences of antimicrobial resistance. AB - BACKGROUND: In the past two decades, a dramatic increase in the frequency and prevalence of antimicrobial-resistant pathogens has challenged clinicians and researchers. MATERIALS AND METHODS: A review of the literature was conducted. Available data identifying the costs and consequences of resistance are summarized while the issues and limitations of research assessing the economics of resistance are acknowledged. RESULTS: Microbial resistance is a complex, multifactorial phenomenon, but the single most powerful influence is antimicrobial use. Treatment guidelines, clinical pathways, and other directives exert widespread influences on individual selection of antimicrobial agents. However, use of an empiric regimen that does not provide effective coverage, or a targeted regimen that is dosed too low to provide optimal therapy, will delay eradication of the pathogen, increase the potential for resistance to emerge, extend and increase morbidity, and expose the patient to an increased risk of mortality. Coincident with these untoward clinical events are economic consequences secondary to increased duration of treatment, and for some, an extended duration of hospitalization. CONCLUSION: Resistant gram-negative and gram-positive bacteria have been associated with increased direct medical costs ranging from several thousand dollars to tens of thousands of dollars per patient. PMID- 12542928 TI - Abdominal emergencies: diagnostic and therapeutic laparoscopy. AB - BACKGROUND: Laparoscopy has been practiced in the management of emergencies resulting from inflammatory conditions, lumenal obstruction, perforation, vascular occlusion, and trauma. This article identifies and discusses controversial areas in the field, in particular surrounding the efficacy, cost effectiveness, and perceived advantages of laparoscopy in the evaluation and treatment of patients with acute abdominal conditions. MATERIALS AND METHODS: Review and commentary on pertinent articles in the English language literature are presented. RESULTS: Prospective randomized trials have been reported in the treatment of some disorders, but a lack of recommendations that are evidence based has hindered more widespread usage of laparoscopy in an emergency setting. In addition, concerns have been raised that the creation of capnoperitoneum in the patient with established peritonitis may be detrimental with respect to potentiation of bacteremia and severe sepsis, and experimental studies have yielded conflicting data in this regard. CONCLUSION: As such issues are resolved, utilization of laparoscopy is likely to increase substantially as expertise is acquired. A minimal-access approach carries less morbidity and may offer other practical advantages in terms of surgical technique and application. When surgical intervention is appropriate, laparoscopy is now preferred for acute biliary disease. Female patients of reproductive age with acute appendicitis may benefit, particularly if there is preoperative diagnostic uncertainty. Selected cases of intestinal obstruction and visceral perforation presenting soon after symptom onset and in whom shock is absent may also be amenable to laparoscopic repair. Its use in the treatment of most trauma patients and patients with generalized peritonitis or hemodynamic instability is not recommended at present. PMID- 12542929 TI - Systemic Apophysomyces elegans after trauma: case report and literature review. AB - We present a case of systemic fungal infection caused by Apophysomyces elegans in a 50-year-old patient who developed a progressive skin lesion after a motor vehicle crash. Histopathological and mycological examination of the surgical sample showed non-septated hyphae characteristic of mucoraceous fungi. Despite extensive surgical debridement, and parenteral administration of amphotericin B, the patient died of multi-organ failure. Autopsy findings suggested systemic involvement. The fungi recovered from culture had non-apophyseal and globose sporangi, and branched sporaniophores and was identified as Apophysomyces elegans. PMID- 12542930 TI - AIDS presenting with cutaneous Kaposi's sarcoma and bacillary angiomatosis in the bone marrow mimicking Kaposi's sarcoma. AB - Kaposi's sarcoma (KS) and bacillary angiomatosis (BA) may be histologically similar. A precise diagnosis is required because of the different management of these diseases. KS or BA involving bone marrow is rare in patients with and without acquired immune deficiency syndrome (AIDS). We report the case of a 40 year-old human immunodeficiency virus (HIV)-positive homosexual male who presented with small KS lesions in the skin and BA in the bone marrow that histologically were similar. Laboratory evaluation revealed anemia and thrombocytopenia; CD4 count was 103/mm3, and the viral load was 750,000 HIV-1 mRNA copies per milliliter in plasma. Bartonella henselae, the etiologic agent of BA, was isolated from a blood culture. DNA sequences of human herpesvirus-8 (HHV 8), the putative etiologic agent of KS, were identified by polymerase chain reaction (PCR) in skin and bone marrow specimens, but antibody anti-HHV-8-encoded protein ORF73, localized signals only in the skin-KS lesion. The patient received clarithromycin and cefotetan for the BA, and antiretroviral therapy for the HIV infection. The skin lesions gradually regressed, the HIV-1 mRNA copy number decreased to less than 400 per milliliter and the CD4 lymphocyte count increased to 665/mm3. In conclusion, vascular lesions of BA and KS may be clinically and histologically similar, both may be associated with advanced AIDS, and an accurate diagnosis is required because of their different management. PMID- 12542931 TI - Vacuolar myelopathy and vacuolar cerebellar leukoencephalopathy: a late complication of AIDS after highly active antiretroviral therapy-induced immune reconstitution. AB - Controversy exists as to whether vacuolar myelopathy (VM) responds to highly active antiretroviral therapy (HAART) in a salutary fashion similar to other primary human immunodeficiency virus (HIV)-related neurologic complications such as acquired immune deficiency syndrome (AIDS) dementia complex and progressive multifocal leukoencephalopathy. Herein, we describe the case of a patient with AIDS, non-Hodgkin's lymphoma, and cytomegalovirus colitis, who began HAART and cytotoxic chemotherapy. After 6 months of therapy, restaging studies showed no residual lymphoma or active opportunistic infection. For 2 years he was maintained on HAART, during which time his HIV viral load remained nondetectable and his CD4+ count improved from 20 to 300 cells per microliter. Shortly after developing the acute onset of cerebellar ataxia, he aspirated, developed adult respiratory distress syndrome, and died. At autopsy the spinal cord demonstrated a characteristic vacuolated appearance that extended into the cerebellum. No relation between HIV and the development of VM was discerned by in situ hybridization studies. Experience with this one patient suggests that HAART may not alter the natural history of VM. Whether this case represents yet another variant of the recently described inflammatory immune response syndrome whereby progression of previously quiescent disorders evolve to symptomatic disease after initiation of HAART is uncertain. PMID- 12542932 TI - Overcoming obstacles to the success of protease inhibitors in highly active antiretroviral therapy regimens. AB - Success with current protease inhibitors (PIs) is limited by substantial variability in pharmacokinetics, onset of adverse metabolic effects that include sustained lipid elevations and insulin resistance, and increased risk of lipodystrophy. Additionally, poor adherence to the often complex regimens can lead to emergence of PI-resistant human immunodeficiency virus (HIV) variants and treatment failure. Boosting blood levels of current PIs through coadministration of ritonavir can improve the pharmacokinetic characteristics of these agents, increasing the chances of success, but often at the price of additional adverse effects. New PIs in development have the potential to overcome at least some of these obstacles. Tipranavir, mozenavir, and atazanavir have favorable and unique resistance profiles, making them potentially effective in new treatment strategies in both PI-naive and PI-experienced patients. Atazanavir does not cause the lipid elevations seen with current PIs, and it may improve adherence through once-daily dosing. PMID- 12542933 TI - Predictors of antiretroviral adherence as measured by self-report, electronic monitoring, and medication diaries. AB - This study reports on the prevalence and predictors of adherence as measured by self-report, medication diaries, and electronic monitoring. A sample of 180 patients receiving human immunodeficiency virus (HIV) antiretroviral therapy were randomized to one of three adherence surveillance methods (electronic monitoring caps, patient medication diaries, no surveillance control group) for 4 weeks, with adherence measured by self-report at study end point; 173 (96%) participants completed the study. Most participants were male (82%), non-white (68%), and unemployed (84%); mean self-reported CD4 count was 406 cells/mm,3 and 31% reported having undetectable viral load. The three methods of adherence measurement revealed moderate (electronic monitoring: 80.6%) to high (self report: 93.7%; diaries: 92.6%) rates of adherence. Although there were differences across adherence measures, variables that were identified as correlates of lower adherence by all three methods included living with others or having a larger household, current symptomatology, and cognitive functioning deficits. This consensus across adherence measures strengthens the validity of these identified predictors and their potential value in informing the development of adherence interventions, as well as increases the validity of the three adherence measures. PMID- 12542935 TI - Antiviral Briefs. PMID- 12542934 TI - Self-reported adherence to antiretroviral treatment and degree of sense of coherence in a group of HIV-infected patients. AB - The goal of this study was to explore the relation between self-reported adherence to antiretroviral treatment and degree of sense of coherence in a group of human immunodeficiency virus (HIV)-infected patients. Ninety-nine patients from an outpatient clinic, all undergoing antiretroviral therapy, participated. Questionnaires were answered twice at a 12-month interval. The 29-item Sense of Coherence (SOC) Scale was used for measuring the ability to cope with stressful life situations. Medication adherence was assessed with self-reported measurements. Clinical characteristics and background variables were collected from the medical records. Results from variables measured with the 12-month interval show a significant concordance with disease stage (p < or = 0.0001), and in HIV-RNA copies per milliliter (p < or = 0.0001) and an increase in CD4 cell count/mm3 (p = < 0.0001). Univariate analysis showed significant differences between nonadherent patients (n = 19) and adherent patients (n = 80) at the last measurement (i.e., the nonadherent group had lower CD4 cell count/mm3 [p = 0.004], higher HIV-1 RNA levels [p 50.029], and lower SOC [p = 0.04] than the adherent group). Finally, multiple regression analyses showed that at measurement 2 the SOC predicted nonadherence, the lower SOC the more missed doses (p < or = 0.01). Because SOC seems to play an important role in this group of patients managing their disease, a caring patient-provider relationship should be developed to minimize nonadherent behaviour. For this reason SOC scale might be of great clinical value to identify patients needing the most support for successful treatment. PMID- 12542936 TI - Replicative aging and gene expression in long-term cultures of human bone marrow stromal cells. AB - Bone marrow stromal cells (BMSCs) can be easily isolated from adult marrow and contain a population of pluripotent progenitors that can give rise to different mesenchymal lineages both in vitro and in vivo. These properties make BMSCs an attractive target for cell-based therapeutic strategies for a variety of disorders. However, because of their low frequency in vivo, to obtain a sufficient number of cells for tissue engineering a step of extensive in vitro expansion is required, which could significantly alter BMSC properties. Therefore, effective therapeutic use of BMSCs requires the design of appropriate approaches for in vitro cell expansion. In this study we have investigated the biological effects of in vitro expansion on BMSC proliferative ability and on their spontaneous differentiation. Telomerase activity and telomere shortening kinetics were evaluated together with variations in osteogenic, chondrogenic, and adipogenic gene expression throughout the BMSC life span. In culture BMSCs never displayed telomerase activity and during in vitro expansion telomere length decreased. Furthermore, gene expression patterns spontaneously varied during expansion, indicating a progressive commitment of the population toward the osteogenic lineage. In conclusion, BMSCs in culture undergo progressive replicative aging and osteogenic differentiation. These observations are relevant to their successful use in clinics and should be considered when designing novel therapeutic strategies. PMID- 12542937 TI - Bone tissue-engineered implants using human bone marrow stromal cells: effect of culture conditions and donor age. AB - At present, it is well known that populations of human bone marrow stromal cells (HBMSCs) can differentiate into osteoblasts and produce bone. However, the amount of cells with osteogenic potential that is ultimately obtained will still be dependent on both patient physiological status and culture system. In addition, to use a cell therapy approach in orthopedics, large cell numbers will be required and, as a result, knowledge of the factors affecting the growth kinetics of these cells is needed. In the present study we investigated the effect of dexamethasone stimulation on the in vivo osteogenic potential of HBMSCs. After a proliferation step, the cells were seeded and cultured on porous calcium phosphate scaffolds for 1 week, and then subcutaneously implanted in nude mice for 6 weeks, in order to evaluate their in vivo bone-forming ability. Furthermore, the effect of donor age on the proliferation rate of the cultures and their ability to induce in vivo bone formation was studied. In 67% of the assayed patients (8 of 12), the presence of dexamethasone in culture was not required to obtain in vivo bone tissue formation. However, in cultures without bone-forming ability or with a low degree of osteogenesis, dexamethasone increased the bone-forming capacity of the cells. During cellular proliferation, a significant age-related decrease was observed in the growth rate of cells from donors older than 50 years as compared with younger donors. With regard to the effect of donor age on in vivo bone formation, HBMSCs from several donors in all age groups proved to possess in vivo osteogenic potential, indicating that the use of cell therapy in the repair of bone defects can be applicable irrespective of patient age. However, the increase in donor age significantly decreased the frequency of cases in which bone formation was observed. PMID- 12542938 TI - Tissue-engineered repair of osteochondral defects: effects of the age of donor cells and host tissue. AB - Transplantation of a tissue-engineered construct containing cells of a chondrocytic phenotype into an osteochondral defect provides a biological solution to this type of injury. Among the factors that affect cell proliferation and phenotypic expression, age is one that has not been well characterized. In this study adult and aged male donor cells, derived from perichondrium, were cultured and adsorbed into a polylactic acid (PLA) scaffold and transplanted into osteochondral defects created in adult (8- to 10-month-old) and aged (4- to 5 year-old) female rabbits. Three groups were investigated: (1) adult cells transplanted into aged defects, (2) aged cells transplanted into aged defects, and (3) aged cells transplanted into adult defects. In vitro characterization of adult and aged cells and in vivo assessment of osteochondral repair tissue at 12 weeks posttransplantation were carried out. The in vitro studies demonstrated that the proliferation rate of aged cells was less than that of mature cells during the earliest stage of culture. Also, the chondrocytic phenotype was reduced in aged cells compared with mature cells. The in vivo results showed that donor (SRY-positive) cell survival differed among the three groups: survival of adult cells into aged defect > survival of aged cells into aged defect > survival of aged cells into adult defect. The biological acceptability of the repair, defined as smooth firm cartilaginous tissue filling the defect, was < 25% of the operated specimens in each of the three groups. This repair tissue contained only 20-25% of the amounts of type II collagen and glycosaminoglycans found in normal articular cartilage. These data suggest that the outcome of tissue-engineered repair of osteochondral defects is affected by both the age of donor cells and the age of the host. PMID- 12542939 TI - Mechanical and chemical characteristics of mineral produced by basic fibroblast growth factor-treated bone marrow stromal cells in vitro. AB - It has been shown that various organ and cell cultures exhibit increased mineral formation with the addition of basic fibroblast growth factor (bFGF) and phosphate ions in the medium. However, to date there has been no attempt to relate the chemical composition of mineral formed in vitro to a measure of its mechanical properties. This information is important for understanding the in vivo mineralization process, the development of in vitro models, and the design of tissue-engineered bone substitutes. In this study we examined the reduced modulus; hardness; and mineral-to-matrix, crystallinity, carbonate-to-mineral, and calcium-to-phosphorus ratios of mineral formed by bFGF-treated rat-derived bone marrow stromal cells in vitro. The cells were treated with 1 or 3 mM beta glycerophosphate for 3 and 4 weeks. Both mechanical parameters, reduced modulus and hardness, increased with increasing beta-glycerophosphate concentration. The only chemical measure of the mineral composition that exhibited the same dependency was the mineral-to-matrix ratio. The values of crystallinity and carbonate fraction were similar to those for intact cortical bone, but the calcium-to-phosphorus ratio was substantially lower than that of normal bone. These data indicate that the mineral formed by bFGF-treated bone cells is mechanically and chemically different from naturally formed lamellar bone tissue after 4 weeks in culture. These results can be used to improve in vitro models of mineral formation as well as enhance the design of tissue-engineered bone substitutes. PMID- 12542940 TI - In vitro control of human bone marrow stromal cells for bone tissue engineering. AB - For the clinical application of cultured human mesenchymal stem cells (MSCs), cells must have minimal contact with fetal calf serum (FCS) because it might be a potential vector for contamination by adventitious agents. The use of human plasma and serum for clinical applications also continues to give rise to considerable concerns with respect to the transmission of known and unknown human infectious agents. With the objective of clinical applications of cultured human MSCs, we tested the ability of autologous plasma, AB human serum, FCS, and artificial serum substitutes containing animal-derived proteins (Ultroser G) or vegetable-derived proteins (Prolifix S6) to permit their growth and differentiation in vitro. To conserve as much autologous plasma as possible, we attempted to mix it at decreasing concentrations with the serum substitute containing vegetable-derived mitogenic factors. Under control conditions, by day 10 all the fibroblast colony-forming units (CFU-Fs) were alkaline phosphatase (ALP) positive. However, their number and size were highly variable among donors. Better CFU-F formation was obtained with Ultroser G, and with human AB serum and autologous plasma mixed at, respectively, 5 and 1% with Prolifix S6. The effects of these mixtures on CFU-F formation demonstrate synergy, with the human serum or plasma supplying the factors that favor differentiation of MSCs while Prolifix S6 supplies the mitogenic factors. Finally, we demonstrated the possibility of controlling human MSC growth and differentiation in vitro. Notably, by means of a minimal quantity of human serum or human plasma mixed with a new serum substitute containing vegetable-derived proteins, we displayed growth and differentiation of human MSCs comparable to that obtained with FCS or serum substitutes containing animal-derived proteins. These results will have crucial significance for future applications of cultured human MSCs in bone tissue engineering. PMID- 12542941 TI - Small intestinal submucosa versus salt-extracted polyglycolic acid-poly-L-lactic acid: a comparison of neocartilage formed in two scaffold materials. AB - This study sought to compare differences in neocartilage produced over time from two types of resorbable scaffold materials. One material was entirely synthetic and contained a polyglycolic acid-poly-L-lactic acid matrix (PGA-PLLA). The second scaffold material was bioactive and consisted of a four-layered construct of porcine small intestinal submucosa (SIS). Disk-shaped scaffolds were seeded with canine chondrocytes and implanted into athymic mice for periods of 5, 8, 12, and 24 weeks. Constructs were examined microscopically, assayed for hydroxyproline (HP) and glycosaminoglycan (GAG) content, and collagen typed (I or II) at each time period. Creep indentation tests determined aggregate and shear modulus, permeability, and thickness. Results indicated that SIS maintained its thickness through the first 12 weeks, and then doubled by week 24. The 24-week tissue appeared chondroid-like and possessed high GAG content. Tissues derived from PGA-PLLA scaffolds were lower in HP content than SIS-derived tissues, but type II collagen was demonstrated only in PGA-PLLA-derived tissues at 24 weeks. Mechanical properties were not significantly different for any tissue over time (p > 0.05), but aggregate and shear modulus mean values were consistently higher for PGA-PLLA-derived tissues at nearly every time interval. This, coupled with the presence of collagen types I and II, suggested a more congruent solid phase may be forming within the extracellular matrix of tissues derived from PGA-PLLA scaffolds. Future study is necessary to compare these materials under simulated loading conditions. PMID- 12542942 TI - Use of controlled mechanical stimulation in vivo to induce cartilage layer formation on the surface of osteotomized bone. AB - A micromachine was used to study the response of mesenchymal tissue to mechanically controlled motion in vivo. The middle portion of the coccygeal vertebra of Fischer 344 rats was osteotomized, and continuous bending motion was applied for 4 weeks. The experimental groups were divided into two groups with higher sliding displacement applied at the osteotomized gap of group II. Hyaline cartilage tissue was generated at the osteotomized ends, and was predominantly formed on the side that extended during the bending motion. These newly formed tissues stained intensively with safranin O and toluidine blue, positively with immunostain for type II collagen, but negatively with immunostain for type I collagen. Articular cartilage-like tissues with a surface and a layer structure were obtained in group II, in which higher sliding motion was applied. Light and electron microscopy revealed morphological features similar to those of normal articular cartilage tissue in the superficial and middle zones of the tissues obtained in group II. Collagen fibrils in the superficial zone were found aligned parallel to the smooth surface. Although tidemark formation was not observed in the deep zone, the structure was much more natural than that of any other tissue engineered cartilage reported to date. These results suggest that controlled sliding stimulation can elicit the generation of articular cartilage structure in vivo. PMID- 12542943 TI - Collagen fibrillogenesis by chondrocytes in alginate. AB - Collagen is the primary structural component in connective tissue. The poor mechanical properties of most cell-seeded cartilage grafts used for cartilage repair can be attributed to the low level of collagen synthesized compared with native cartilage. In this study, the synthesis and assembly of collagen by chondrocytes in hydrogels were investigated, with particular attention paid to the role of cross-link formation in this process. Primary bovine chondrocytes were seeded in alginate and collagen synthesis was assessed in the presence and absence of beta-aminopropronitrile (BAPN), a potent inhibitor of the enzyme lysyl oxidase and collagen cross-link formation. Cultures on days 21, 35, and 49 were evaluated by stereology, biochemistry, and real-time reverse transcriptase polymerase chain reaction. All measures of collagen synthesis (except hydroxyproline) significantly increased in the presence of 0.25 mM BAPN. By 35 days of culture, the average collagen fibril diameter was 62 +/- 10 nm in control cultures and 109 +/- 20 nm with BAPN supplementation. The collagen volume density increased from 5 +/- 3% in control cultures to 17 +/- 1% in the presence of BAPN. Likewise, the expression of cartilage-specific collagens (type II and XI) and aggrecan increased significantly as a result of BAPN culture. These findings demonstrate the prominent role of collagen cross-linking in collagen fibrillogenesis and suggest approaches by which collagen synthesis and assembly could be controlled in tissue-engineered constructs. PMID- 12542944 TI - Interfibrillar collagen bonding exists in matrix produced by chondrocytes in culture: evidence by electron microscopy. AB - Interfibrillar bonding of collagen fibrils in tissue grown from rabbit chondrocytes in culture was examined by a variety of electron microscopy techniques. Interfibrillar bonding is expected to increase tissue strength and may be a desirable feature in engineered cartilage and other soft tissues. The apparent bonding evident by scanning electron microscopy, using standard chemical fixation processing, is suspected to be artifact due to drying. The goal of this article was to establish the existence of interfibrillar bonding, apart from any processing artifacts. Specimens prepared by transmission electron microscopy, scanning electron microscopy (SEM) after notching and fixing under load, and cryo SEM all showed evidence of bonding, supporting the existence of bonding in the unprocessed tissue. Exclusion from the bond space of gold particles labeled to decorin further supported the existence of natural bonds. Artifactual bonding may still be occurring with some of the methods used, but interfibrillar bonds exist in natural tissue. The bond distance was estimated to be 7-14 nm. Demonstration of the existence of these bonds supports further study of their mechanism and effect on tissue properties. PMID- 12542945 TI - Engineering a bioactive matrix by modifications of calcium sulfate. AB - The goal of this study was to define the conditions for the fabrication of a bioactive matrix that induces and supports cell proliferation and tissue regeneration. The proposed hypothesis was that a composite graft could be engineered by the absorption of platelet-rich plasma (PRP) onto calcium sulfate (CS). Evaluation of the biological activity of the engineered grafts was based on osteoblast proliferation studies and scanning electron microscopy (SEM) analyses. Graft samples were created in a standard size and shape so that the surface available for attachment and cell proliferation was always identical. Proliferation data were expressed as counts per minute per group and differences among groups were statistically analyzed by analysis of variance followed by the Scheffe test (alpha = 0.1). SEM analysis showed that the combination of CS and PRP presents a preserved crystalline structure well integrated by organic matrix. This combination showed the highest cell proliferation levels (p < 0.001). Further evaluations demonstrated that PRP is activated when combined with CS. When tested as a possible carrier for biologically active molecules such as platelet-derived growth factor (PDGF), CS showed increased cell proliferation (p < 0.001). SEM revealed adherent osteoblasts with broad flattened edges on CS-PRP. This study proposes CS as an efficient carrier for PRP or PDGF and supports the use of these combinations as bioactive matrices in clinical or laboratory applications. PMID- 12542946 TI - Potential use of chitosan as a cell scaffold material for cartilage tissue engineering. AB - One of the most important factors in any tissue-engineering application is the cell substrate. The purpose of this study was the initial evaluation of chitosan, a derivative of the abundant, naturally occurring biopolymer chitin, as a cell scaffold for cartilage tissue engineering. Chitosan scaffolds having an interconnecting porous structure were easily fabricated by simple freezing and lyophilization of a chitosan solution. After rehydration of scaffolds, porcine chondrocytes were seeded onto scaffolds and cultured for up to 28 days in a rotating-wall bioreactor. Chitosan scaffolds supported cell attachment and maintenance of a rounded cell morphology. After 18 days, cells within the scaffolds had synthesized extracellular matrix in which proteoglycan and type II collagen were detected by toluidine blue staining and immunohistochemistry, respectively. Abundant extracellular matrix was found almost exclusively in the periphery of the scaffolds, as scaffold microstructure prevented cells from penetrating to interior regions. Nonetheless, the results suggest that chitosan scaffolds may be a useful alternative to synthetic cell scaffolds for cartilage tissue engineering. PMID- 12542947 TI - Porous poly(propylene fumarate) foam coating of orthotopic cortical bone grafts for improved osteoconduction. AB - A porous biodegradable scaffold coating for perforated and demineralized cortical bone allografts could maintain immediate structural recovery and subsequently allow normal healing and remodeling by promoting bony ingrowth and avoiding accelerated graft resorption. This new type of osteoconductive surface modification should improve allograft incorporation by promoting new bone growth throughout the biodegradable scaffold, hence encasing the graft with the recipient's own bone. We investigated the feasibility of augmenting orthotopically transplanted cortical bone grafts with osteoconductive biodegradable polymeric scaffold coatings. Five types of bone grafts were prepared: type I, untreated fresh-frozen cortical bone grafts (negative control); type II, perforated and partially demineralized cortical bone grafts without additional coating (positive control); type III, perforated and partially demineralized cortical bone coated with a low-porosity poly(propylene fumarate) (PPF) foam; type IV, perforated and partially demineralized cortical bone coated with a medium-porosity PPF foam; and type V, perforated and partially demineralized cortical bone coated with a high-porosity PPF foam. Grafts were implanted into the rat tibial diaphysis. Fixation was achieved with an intramedullary threaded K-wire. Two sets of animals were operated on. Animals were killed in groups of eight with one set being killed 12 weeks, and the other 16 weeks, postoperatively. Radiographic, histologic, and histomorphometric analyses of grafts showed that the amount of new bone forming around the foam coated grafts was significantly higher than that in the type I control group (uncoated) or that in type II group (perforated and partially demineralized cortical bone grafts). Although all foam formulations appeared initially equally osteoconductive, histologic evaluation of medium-porosity PPF foam-based coatings appeared to result in a sustained response 16 weeks postoperatively. Significant resorption was present in perforated and partially demineralized cortical bone graft allografts, with some accompanying new bone formation occurring primarily within the laser holes. Therefore, PPF foam-coated cortical bone grafts appeared to be better protected from excessive bone resorption, as frequently seen with invasion of fibrovascular tissue. Biomechanical analysis of the PPF foam-coated grafts corroborated findings of the morphometric analysis in that the failure strength at the allograft-host bone junction sites of all PPF-coated cortical bone grafts was higher than in the uncoated controls. PMID- 12542948 TI - In vivo magnetic resonance imaging explorative study of ectopic bone formation in the rat. AB - In animal studies of tissue engineering of bone, histology remains the standard for assessing bone formation. As longitudinal studies with this method are feasible only at the cost of large numbers of animals, we looked for an alternative. Therefore, demineralized bone matrix (DBM) and inactivated demineralized bone matrix (iDBM) implants were subcutaneously implanted in a rat. At 1, 3, 5, and 7 weeks postimplantation soft X-ray and magnetic resonance imaging (MRI) were done to monitor bone formation in the implants. At 7 weeks, the animal was killed and the implants were retrieved for histology. Our results showed that in vivo MRI is well suited to assess bone formation larger than 0.5 mm in diameter and to monitor the complete three-dimensional shape of the newly formed bone noninvasively and longitudinally. The MRI results matched well with the histology results obtained at 7 weeks. In contrast, X-ray imaging appeared inappropriate to monitor the bone formation process in DBM. PMID- 12542949 TI - Proteoglycans synthesized by canine intervertebral disc cells grown in a type I collagen-glycosaminoglycan matrix. AB - The objective of this study was to determine the characteristics of proteoglycans synthesized by canine annulus fibrosus cells expanded in number in monolayer culture through passage 4 and subsequently grown in a type I collagen glycosaminoglycan matrix to be employed for tissue engineering. Newly synthesized [35S]sulfate-labeled proteoglycans were analyzed by gel chromatography, including sequential digestion with enzymes and nitrous acid. After 1 week in culture, the percentage of cell-associated, aggregated proteoglycans synthesized in type I collagen-glycosaminoglycan matrices was 52% compared with 38% by the cells in monolayer. The percentage of aggregated proteoglycan in each group increased only slightly with the addition of exogenous hyaluronic acid, but remained significantly different from each other. There were at least three different hydrodynamic sizes of proteoglycans both in the collagen-glycosaminoglycan matrix and in monolayer; the average size was larger in the collagen matrices and the glycosaminoglycan chains were longer. The proteoglycans contained chondroitin sulfate, dermatan sulfate, heparan sulfate, and keratan sulfate. The results provide a foundation for future investigations of collagen-glycosaminoglycan matrices for intervertebral disc tissue engineering. PMID- 12542950 TI - Ultrastructure of proteoglycans in tissue-engineered cardiovascular structures. AB - Proteoglycans such as versican, decorin, and perlecan are important components of the extracellular matrix in various tissues. They play an important role in water homeostasis, tissue elasticity, prevention of calcification, and thrombogenicity. The aim of our study was to detect such proteoglycans in engineered tissue and compare them with the proteoglycans of native porcine heart valves. Myofibroblasts were seeded on a type I collagen scaffold. Thereafter, endothelial cells were seeded onto the presettled myofibroblasts. The newly formed tissue was histologically and immunohistochemically examined. Cupromeronic blue was used for ultracytochemical staining of proteoglycans. Radiolabeled proteoglycans were isolated by ion-exchange chromatography and characterized by enzymatic degradation. Three differently sized proteoglycan precipitates were found. The large-sized proteoglycan (154 nm) was located outside the collagen bundles in a rarely structured extracellular matrix compound. The small-sized proteoglycan (46 nm) was aligned along the collagen bundles at intervals of 60 nm. The intermediate-sized proteoglycan (56 nm) was detected on the cell surface of myofibroblasts. The glycosaminoglycans included 80% chondroitin and dermatan sulfate and 20% heparan sulfate. We conclude that proteoglycans play an important role in the functional integrity of cardiovascular tissues. This study shows the successful production of a heart valve-like tissue with proteoglycans resembling, in terms of type, production, and distribution, proteoglycans of native heart valves. PMID- 12542951 TI - Systemic delivery of human growth hormone using genetically modified tissue engineered microvascular networks: prolonged delivery and endothelial survival with inclusion of nonendothelial cells. AB - Endothelial cells have the potential to provide efficient long-term delivery of therapeutic proteins to the circulation if a sufficient number of genetically modified endothelial cells can be incorporated into the host vasculature and if these cells persist for an adequate period of time. Here we describe the ability of nonendothelial cells to modulate the survival of implanted endothelial cells and their incorporation into host vasculature. Bovine aortic endothelial cells (BAECs) suspended in Matrigel and cultured in vitro remained spherical and decreased in number over time. Subcutaneous implantation of gels containing BAECs secreting human growth hormone (hGH) in mice initially resulted in detectable plasma hGH levels, which were undetectable after 2 weeks. When mixed with fibroblasts and suspended in Matrigel, hGH-secreting BAECs formed microvascular networks in vitro. Implantation of these gels resulted in plasma hGH levels that decreased slightly over 2 weeks and then remained stable for at least 6 weeks. BAECs incorporated into blood vessels within both the implant and fibrous capsule that surrounded and invaded implants. Within implants containing BAECs and fibroblasts, viable BAECs were present for at least 6 weeks at a higher density than in implants containing BAECs alone at 3 weeks. These results indicate that implanted BAECs can incorporate into host blood vessels and that inclusion of fibroblasts in this system prolongs BAEC survival and hGH delivery. PMID- 12542952 TI - Differentiation kinetics of in vitro 3T3-L1 preadipocyte cultures. AB - Engineering autologous adipose constructs from cell culture is a promising strategy to overcome limitations of conventional soft-tissue implants. A methodology is presented to experimentally determine and mathematically model the differentiation kinetics of in vitro 3T3-L1 preadipocyte cultures that can aid in construct design. Relative rates of morphological and interfacial events during adipogenesis were compared. Model results suggest that maturation of an intermediate multilocular phenotype was the rate-limiting step in morphological differentiation and had an intrinsic rate of 0.012 day(-1). Dislodgment of multilocular fat cells was the primary mechanism of cell loss during adipogenesis. The maximum rate of lipid droplet nucleation was predicted to precede that of coalescence by 10 days and to be three times faster. Coalescence probability was estimated to decrease from 33 to 11% for 4- and 8-microm-diameter droplets, respectively. Fluid drainage and the cytoskeleton between droplets could have impeded coalescence. The kinetic analysis suggests that droplet ripening was the dominant mechanism of lipid production. Applications of this research include engineering of an adipose construct and predicting surgical outcome of patients requiring soft-tissue augmentation. PMID- 12542953 TI - A novel pulsatile, laminar flow bioreactor for the development of tissue engineered vascular structures. AB - Exposure of vascular cell-seeded, tubular, biodegradable polymers to pulsatile flow conditions has been proposed as a method to develop tissue-engineered blood vessels by "maturing" structural integrity, and increasing collagen content, suture retention, burst pressure, and tissue formation. These in vitro tissue engineered arteries demonstrate contractile responses to pharmacologic agents and express markers of vascular differentiation. Current methods to induce pulsatile flow in a bioreactor system are limited by the creation of nonphysiologic pressure waveforms and noncompliant reservoirs to house the tissue-engineered vascular constructs. We have developed a novel method for the in vitro development of tubular vascular structures by using a mechanical ventilator to induce pulsatile, laminar flow into a fluid column, resulting in pressurized waveforms similar to mammalian physiology. The vascular constructs are housed in semicompliant tubing to facilitate an additional variable of circumferential stretch as a potential signaling mechanism. This approach more closely approximates mammalian physiology and we hypothesize that it will facilitate mechanical signaling necessary for the development of tissue-engineered vessels for clinical applications. PMID- 12542954 TI - Microsyringe-based deposition of two-dimensional and three-dimensional polymer scaffolds with a well-defined geometry for application to tissue engineering. AB - A technique for controlled deposition of biomaterials and cells in specific and complex architectures is described. It employs a highly accurate three dimensional micropositioning system with a pressure-controlled syringe to deposit biopolymer structures with a lateral resolution of 5 microm. The pressure activated microsyringe is equipped with a fine-bore exit needle and a wide variety of two- and three-dimensional patterns on which cells to be deposited can adhere. The system has been characterized in terms of deposition parameters such as applied pressure, motor speed, line width and height, and polymer viscosity, and a fluid dynamic model simulating the deposition process has been developed, allowing an accurate prediction of the topological characteristics of the polymer structures. PMID- 12542955 TI - Polymer-demixed nanotopography: control of fibroblast spreading and proliferation. AB - Cell response to nanometric scale topography is a growing field. Nanometric topography production has traditionally relied on expensive and time-consuming techniques such as electron beam lithography. This presents disadvantages to the cell biologist in regard to material availability. New research is focusing on less expensive methods of nanotopography production for in vitro cell engineering. One such method is the spontaneous demixing of polymers (in this case polystyrene and polybromostyrene) to produce nanometrically high islands. This article observes fibroblast response to nanometric islands (13, 35, and 95 nm in height) produced by polymer demixing. Changes in cell morphology, cytoskeleton, and proliferation are observed by light, fluorescence, and scanning electron microscopy. Morphological features produced by cells in response to the materials were selected, and cell shape parameters were measured with shape recognition software. The results showed that island height could either increase or reduce cell spreading and proliferation in relation to control, with 13-nm islands producing cells with the greatest area and 95 nm islands producing cells with the lowest areas. Interaction of filopodia with the islands could been seen to increase as island size was increased. PMID- 12542956 TI - Reciprocal action between BMP-2 and BMP-3 in cultured fibroblast in vitro. AB - OBJECTIVE: To explore reciprocal action between BMP-2 (bone morphogenetic protein 2) and BMP-3 for better understanding of the mechanism of BMP during bone fracture union. METHODS: rhBMP-2 was added into the cultured fibroblasts with the concentration of 1,200 ng/ml. The expression of BMP-3 in fibroblasts was detected by immunohistochemistry. Eukaryotic expression vector pcDNA3-BMP-3 was transfected into the fibroblasts. After the effective expression of BMP-3 was identified, BMP-2 was also detected by immunohistochemistry in BMP-3 expression cells. The fibroblasts transfected with empty vector pcDNA3 were used as the control. RESULTS: Exogenous rhBMP-2 could promote the expression of BMP-3 in fibroblasts. BMP-3 also could be detected in these cells. CONCLUSIONS: BMP-2 and BMP-3 could reciprocally adjust the expression in fibroblasts. PMID- 12542957 TI - Thermal stress can inhibit proliferation of ECV304 cells. AB - OBJECTIVE: To observe the effects of thermal stress on proliferation of human vascular endothelial cells (VECs) and explore its significance. METHODS: Changes of VECs proliferation were investigated with (3)H-TdR incorporation method after ECV304 was treated at 43 degrees for 2 hours, while expressions of intercellular adhesion molecule-1 (ICAM-1), inhibitor of differentiation-1 (ID1), and P16 and P21 proteins were determined by Western Blotting. RESULTS: The effect of inhibition of VECs growth after thermal stress was detected by (3)H-TdR incorporation experiment. Western blotting showed ICAM-1, a marker of activated endothelial cells, was increased markedly after thermal stress. Expression of ID1 protein declined gradually with increasing expressions of its downstream genes, P16 and P21 following the thermal stress. CONCLUSIONS: Thermal stress could strongly activate VECs and inhibit proliferation of VECs through ID1, thus down regulating cyclin-dependent kinase inhibitors, P16 and P21, which might be an essential pathway for recovery of VECs after thermal stress. PMID- 12542958 TI - Risk factors of military training-related injuries in recruits of Chinese People's Armed Police Forces. AB - OBJECTIVE: To assess the incidence, types and risk factors of military training related injuries in recruits of Chinese People's Armed Police Forces (CPAPF). METHODS: A cohort study was made on the risk factors of injuries in 805 male recruits during the military training from December 25, 1999 to December 25, 2000. RESULTS: A total of 111 recruits (14%) experienced one or more injuries, and the cumulative incidence was 16.1 injuries per 100 soldiers in a year. And 77.7% of the injuries belonged to overuse injuries of the skeletal and muscular systems, the most common type of which was stress fractures. Most injuries occurred in the 3rd month of training. Univariate analysis and logistic regression analysis of possible risk factors for overuse injuries were carried out, and a number of risk factors were identified: history of agricultural labor, history of lower limb injury, flatfoot and less running exercise before entry into the army. But a suitable body mass index (BMI) was a protective factor. Examination of age, body height, smoking, body flexibility and frequency of 2 mile running revealed no significant association with the injuries. CONCLUSIONS: History of agricultural labor, history of lower limb injury, flatfoot, less running exercise before entry into the army and lower BMI were risk factors of the overuse injuries. In order to decrease the incidence of overuse injuries, the young people with good physical ability and shapely body type should be selected during conscription. During the training, nutrition should be improved so as to decrease the incidence of injuries. PMID- 12542959 TI - Detection of gene expression pattern in the early stage after spinal cord injury by gene chip. AB - OBJECTIVE: To study the changes of the gene expression pattern of spinal cord tissues in the early stage after injury by DNA microarray (gene chip). METHODS: The contusion model of rat spinal cord was established according to Allen's falling strike method and the gene expression patterns of normal and injured spinal cord tissues were studied by gene chip. RESULTS: The expression of 45 genes was significantly changed in the early stage after spinal cord injury, in which 22 genes up-regulated and 23 genes down-regulated. CONCLUSIONS: The expression of some genes changes significantly in the early stage after spinal cord injury, which indicates the complexity of secondary spinal cord injury. PMID- 12542960 TI - Effects of ganglioside GM1 on reduction of brain edema and amelioration of cerebral metabolism after traumatic brain injury. AB - OBJECTIVE: To observe the effects of ganglioside GM1 on reduction of brain edema and amelioration of cerebral metabolism after traumatic brain injury (TBI). METHODS: An acute experimental closed TBI model in rats was induced by a fluid percussion brain injury model. At five and sixty minutes after TBI, the animals were intraperitoneally injected by ganglioside GM1 (30 mg/kg) or the same volume of saline. At the 6th hour after TBI, effects of ganglioside GM1 or saline on changes of mean arterial pressure (MAP), contents of water, lactic acid (LA) and lipid peroxidation (LPO) in the injured cerebral tissues were observed. RESULTS: After TBI, MAP decreased and contents of water, LA and LPO increased in brain injury group; however, MAP was back to normal levels and contents of water, LA and LPO decreased in ganglioside GM1 treated group, compared with those in brain injury group (P < 0.05). No significant difference between the saline treated group and the brain injury group (P > 0.05) was observed. CONCLUSIONS: Ganglioside GM1 does have obvious neuroprotective effect on early TBI. PMID- 12542961 TI - The risk factors of nosocomial infection in severe craniocerebral trauma. AB - OBJECTIVE: To explore the risk factors of nosocomial infection in severe craniocerebral trauma and the way of prevention. METHODS: The clinical data of 387 patients with severe craniocerebral trauma were reviewed. RESULTS: The total nosocomial infection rate of this study was 22.99%. Pulmonary nosocomial infection presented most frequently. The G-bacilli were the most common infectious bacteria. The mortality rate of the infection group was 38.20%. CONCLUSIONS: Complications of nosocomial infection affect the prognosis of craniocerebral trauma patients. Nosocomial infection is related to the age of the patients, craniocerebral trauma severity, unreasonable utilization of antibiotics and invasive operations, such as tracheal cannula, mechanical ventilation, urethral catheterization and deep venous catheterization. Patients with severe craniocerebral trauma should be carefully treated and nursed to avoid nosocomial infection. In order to reduce the rate of nosocomial infection, intensive measurement should be adopted. PMID- 12542962 TI - Component changes of calcium and phosphorus in osteogenesis by lengthening procedures in adult canines. AB - OBJECTIVE: To explore chemical component changes of dog bone at different lengthening time and in different bone regions of interest, and to evaluate the mineralization during Ilizarov lengthening process. METHODS: The ash weight, the concentrations of calcium, phosphorus and the calcium/phosphorus ratio were measured at different intervals (2, 4, 6, 8, 12 weeks) since lengthening and the lengthened part was compared with a control area at each interval. RESULTS: The ash weight, the concentrations of calcium and phosphorus in the lengthened area differed at all development time. The calcium/phosphorus (Ca/P) ratio in the lengthened region remained significantly lower than that in the control region up to 12 weeks after the lengthening. CONCLUSIONS: These results suggest that also other inorganic ions play an important role in the mineralization process and that they become relatively more important since 8 weeks after the lengthening. PMID- 12542963 TI - Animal study on expression of laminin and fibronectin in cornea during wound healing following alkali burn. AB - OBJECTIVE: To observe the expression of laminin and fibronectin in alkali-burned corneas in rats. METHODS: A total of 18 normal Wistar rats were randomly divided into 6 groups (n = 3 in each group). For each rat, one eye was injured by alkali burn, the other one was taken as the normal control. Then all the corneas were surgically removed and the expression of laminin and fibronectin was observed with immunohistochemistry respectively at 7 hours, 1 day, 3 days, 7 days, 14 days and 28 days after alkali burn. RESULTS: Compared with that of the normal controls, the expression of laminin and fibronectin of the burned eyes was dramatically higher at 7 hours, reached peak at 14 days and decreased to the normal level at 28 days after alkali burn. CONCLUSIONS: In the process of wound healing after alkali burn, the expression of laminin and fibronectin increases dramatically, which suggests that laminin and fibronectin may participate in the process of corneal wound healing. PMID- 12542964 TI - Advances in surgical treatment of chronic subdural hematoma. PMID- 12542965 TI - The development of foot microsurgery: the past and the future. PMID- 12542966 TI - Blunt cardiac rupture in patient with liver laceration. PMID- 12542967 TI - Diagnosis and treatment of traumatic hematoma of posterior fossa. PMID- 12542969 TI - A critical evaluation of DNA adducts as biological markers for human exposure to polycyclic aromatic compounds. AB - The causative role of polycyclic aromatic hydrocarbons (PAH) in human carcinogenesis is undisputed. Measurements of PAH-DNA adduct levels in easily accessible white blood cells therefore represent useful early endpoints in exposure intervention or chemoprevention studies. The successful applicability of DNA adducts as early endpoints depends on several criteria: i. adduct levels in easily accessible surrogate tissues should reflect adduct levels in target tissues, ii. toxicokinetics and the temporal relevance should be properly defined. iii. sources of interand intra-individual variability must be known and controllable, and finally iv. adduct analyses must have advantages as compared to other markers of PAHexposure. In general, higher DNA adduct levels or a higher proportion of subjects with detectable DNA adduct levels were found in exposed individuals as compared with nonexposed subjects, but saturation may occur at high exposures. Furthermore, DNA adduct levels varied according to changes in exposure, for example smoking cessation resulted in lower DNA adduct levels and adduct levels paralleled seasonal variations of air-pollution. Intraindividual variation during continuous exposure was low over a short period of time (weeks), but varied significantly when longer time periods (months) were investigated. Inter-individual variation is currently only partly explained by genetic polymorphisms in genes involved in PAH-metabolism and deserves further investigation. DNA adduct measurements may have three advantages over traditional exposure assessment: i. they can smooth the extreme variability in exposure which is typical for environmental toxicants and may integrate exposure over a longer period of time. Therefore, DNA adduct assessment may reduce the monitoring effort. ii. biological monitoring of DNA adducts accounts for all exposure routes. iii. DNA adducts may account for inter-individual differences in uptake, elimination, distribution, metabolism and repair amongst exposed individuals. In conclusion, there is now a sufficiently large scientific basis to justify the application of DNA adduct measurements as biomarkers in exposure assessment and intervention studies. Their use in risk-assessment, however, requires further investigation. PMID- 12542970 TI - Imidazole ring-opened DNA purines and their biological significance. AB - Fragmentation of purine imidazole ring and production of formamidopyrimidines in deoxynucleosides (Fapy lesions) occurs upon DNA oxidation as well as upon spontaneous or alkali-triggered rearrangement of certain alkylated bases. Many chemotherapeutic agents such as cyclophosphamide or thiotepa produce such lesions in DNA. Unsubstituted FapyA and FapyG, formed upon DNA oxidation cause moderate inhibition of DNA synthesis, which is DNA polymerase and sequence dependent. Fapy 7MeG, a methylated counterpart of FapyG-, a efficiently inhibits DNA replication in vitro and in E.coli, however its mutagenic potency is low. This is probably due to preferential incorporation of cytosine opposite Fapy-7MeG and preferential extension of Fapy-7MeG:C pair. In contrast, FapyA and Fapy-7MeA possess miscoding potential. Both lesions in SOS induced E.coli preferentially mispair with cytosine giving rise to A-->G transitions. Fapy lesions substituted with longer chain alkyl groups also show simult aneous lethal and mutagenic properties. Fapy lesions are actively eliminated from DNA by repair glycosylases specific for oxidized purines and pyrimidines both in bacteria and eukaryotic cells. Bacterial enzymes include E.coli formamidopyrimidine-DNA-glycosylase (Fpg protein), endonuclease III (Nth protein) and endonuclease VIII (Nei protein). PMID- 12542972 TI - Genetic polymorphisms and cancer susceptibility of breast cancer in Korean women. AB - Breast cancer is the most prevalent cancer among women in Western countries, and its prevalence is also increasing in Asia. The major risk factor for breast cancer can be traced to reproductive events that influence the lifetime levels of hormones. However, a large percentage of breast cancer cases cannot, be explained by these risk factors. The identification of susceptibility factors that predispose individuals to breast cancer (for instance, if they are exposed to particular environmental agents) could possibly give further insight into the etiology of this malignancy and provide targets for the future development of therapeutics. The most interesting candidate genes include those that mediate a range of functions. These include carcinogen metabolism, DNA repair, steroid hormone metabolism, signal transduction, and cell cycle control. we conducted a hospital-based case-control study on South Korea to evaluate the potential modifying role of the genetic pollymprphisms of selected low penetrance gens that are involved carcinogen metabolisms (i.e., CYP1A1, CYP2E1, GSTM1/T1/P1, NAT1/2, etc.), estrogen synthesis and metabolism (i.e., CYP19, CYP17, CYP1B1, COMT, ER alpha, etc.), DNA repair (i.e., XRCC1/3, ERCC2/4, ATM, AGT, etc.), and signal transduction as well as others (i.e., TGF- beta, IGF-1, TNF- beta, IL-1B, IL-1RN, etc.). We also took into account the potential interaction between these and the known risk factors of breast cancer. The results of selected genes will be presented in this mini-review. PMID- 12542971 TI - Activation of dihaloalkanes by thiol-dependent mechanisms. AB - Dihaloalkanes constitute an important group of chemicals because of their widespread use in industry and agriculture and their potential for causing toxicity and cancer. Chronic toxic effects are considered to depend upon bioactivation, either by oxidation or thiol conjugation. Considerable evidence links genotoxicity and cancer with glutathione conjugations reactions, and some aspects of the mechanisms have been clarified with 1,2-dihaloalkanes and dihalomethanes. Recently the DNA repair protein O6-alkylguanine transferase has been shown to produce cytotoxicity and genotoxicity by means of a thiol-dependent process with similarities to the glutathione reactions. PMID- 12542973 TI - Use of transgenic and mutant animal models in the study of heterocyclic amine induced mutagenesis and carcinogenesis. AB - Heterocyclic amines (HCAs) are potent mutagens generated during the cooking of meat and fish, and several of these compounds produce tumors in conventional experimental animals. During the past 5 years or so, HCAs have been tested in a number of novel in vivo murine models, including the following: lacZ, lacI, cII, c-myc/lacZ, rpsL, and gptDelta. transgenics, XPA-/-, XPC-/-, Msh2+/-, Msh2-/- and p53+/- knock-outs, Apc mutant mice (ApcDelta716, Apc1638N, Apcmin), and A33DeltaNbeta-cat knock-in mice. Several of these models have provided insights into the mutation spectra induced in vivo by HCAs in target and non-target organs for tumorigenesis, as well as demonstrating enhanced susceptibility to HCA induced tumors and preneoplastic lesions. This review describes several of the more recent reports in which novel animal models were used to examine HCA-induced mutagenesis and carcinogenesis in vivo, including a number of studies which assessed the inhibitory activities of chemopreventive agents such as 1,2-dithiole 3-thione, conjugated linoleic acids, tea, curcumin, chlorophyllin-chitosan, and sulindac. PMID- 12542974 TI - The role of stem cells and gap junctional intercellular communication in carcinogenesis. AB - Understanding the process of carcinogenesis will involve both the accumulation of many scientific facts derived from molecular, biochemical, cellular, physiological, whole animal experiments and epidemiological studies, as well as from conceptual understanding as to how to order and integrate those facts. From decades of cancer research, a number of the "hallmarks of cancer" have been identified, as well as their attendant concepts, including oncogenes, tumor suppressor genes, cell cycle biochemistry, hypotheses of metastasis, angiogenesis, etc. While all these "hallmarks" are well known, two important concepts, with their associated scientific observations, have been generally ignored by many in the cancer research field. The objective of the short review is to highlight the concept of the role of human adult pluri-potent stem cells as "target cells" for the carcinogenic process and the concept of the role of gap junctional intercellular communication in the multi-stage, multi-mechanism process of carcinogenesis. With these two concepts, an attempt has been made to integrate the other well-known concepts, such as the multi-stage, multimechanisn or the "initiation/promotion/progression" hypothesis; the stem cell theory of carcinogenesis; the oncogene/tumor suppression theory and the mutation/epigenetic theories of carcinogenesis. This new "integrative" theory tries to explain the well-known "hallmarks" of cancers, including the observation that cancer cells lack either heterologous or homologous gap junctional intercellular communication whereas normal human adult stem cells do not have expressed or functional gap junctional intercellular communication. On the other hand, their normal differentiated, non-stem cell derivatives do express connexins and express gap junctional intercellular communication during their differentiation. Examination of the roles of chemical tumor promoters, oncogenes, connexin knock-out mice and roles of genetically-engineered tumor and normal cells with connexin and anti sense connexin genes, respectively, seems to provide evidence which is consistent with the roles of both stem cells and gap junctional communication playing a major role in carcinogenesis. The integrative hypothesis provides new strategies for chemoprevention and chemotherapy which focuses on modulating connexin gene expression or gap junctional intercellular communication in the premalignant and malignant cells, respectively. PMID- 12542975 TI - Platelet-derived growth factor signaling and human cancer. AB - Platelet-derived growth factor (PDGF) is a critical regulator of mesenchymal cell migration and proliferation. The vital functions of PDGFs for angiogenesis, as well as development of kidney, brain, cardiovascular system and pulmonary alveoli during embryogenesis, have been well demonstrated by gene knock-out approaches. Clinical studies reveal that aberrant expression of PDGF and its receptor is often associated with a variety of disorders including atherosclerosis, fibroproliferative diseases of lungs, kidneys and joints, and neoplasia. PDGF contributes to cancer development and progression by both autocrine and paracrine signaling mechanisms. In this review article, important features of the PDGF isoforms and their cell surface receptor subunits are discussed, with regards to signal transduction, PDGF-isoform specific cellular responses, and involvement in angiogensis, and tumorstromal interactions. PMID- 12542976 TI - Cell cycle and cancer. AB - Cancer is frequently considered to be a disease of the cell cycle. As such, it is not surprising that the deregulation of the cell cycle is one of the most frequent alterations during tumor development. Cell cycle progression is a highlyordered and tightly-regulated process that involves multiple checkpoints that assess extracellular growth signals, cell size, and DNA integrity. Cyclin dependent kinases (CDKs) and their cyclin partners are positive regulators or accelerators that induce cell cycle progression; whereas, cyclindependent kinase inhibitors (CKIs) that act as brakes to stop cell cycle progression in response to regulatory signals are important negative regulators. Cancer originates from the abnormal expression or activation of positive regulators and functional suppression of negative regulators. Therefore, understanding the molecular mechanisms of the deregulation of cell cycle progression in cancer can provide important insights into how normal cells become tumorigenic, as well as how new cancer treatment strategies can be designed. PMID- 12542978 TI - Current mechanistic approaches to the chemoprevention of cancer. AB - The prevention of cancer is one of the most important public health and medical practices of the 21st century. We have made much progress in this new emerging field, but so much remains to be accomplished before widespread use and practice become common place. Cancer chemoprevention encompasses the concepts of inhibition, reversal, and retardation of the cancer process. This process, called carcinogenesis, requires 20-40 years to reach the endpoint called invasive cancer. It typically follows multiple, diverse and complex pathways in a stochastic process of clonal evolution. These pathways appear amenable to inhibition, reversal or retardation at various points. We must therefore identify key pathways in the evolution of the cancer cell that can be exploited to prevent this carcinogenesis process. Basic research is identifying many genetic lesions and epigenetic processes associated with the progression of precancer to invasive disease. Many of these early precancerous lesions favor cell division over quiescence and protect cells against apoptosis when signals are present. Many oncogenes are active during early development and are reactivated in adulthood by aberrant gene promoting errors. Normal regulatory genes are mutated, making them insensitive to normal regulatory signals. Tumor suppressor genes are deleted or mutated rendering them inactive. Thus there is a wide range of defects in cellular machinery which can lead to evolution of the cancer phenotype. Mistakes may not have to appear in a certain order for cells to progress along the cancer pathway. To conquer this diverse disease, we must attack multiple key pathways at once for a predetermined period of time. Thus, agent combination prevention strategies are essential to decrease cancer morbidity. Furthermore, each cancer type may require custom combination of prevention strategies to be successful. PMID- 12542977 TI - Signal transduction pathways: targets for green and black tea polyphenols. AB - Tea is one of the most popular beverages consumed in the world and has been demonstrated to have anti-cancer activity in animal models. Research findings suggest that the polyphenolic compounds, (-)-epigallocatechin-3-gallate found primarily in green tea, and theaflavin-3,3'-digallate, a major component of black tea, are the two most effective anti-cancer factors found in tea. Several mechanisms to explain the chemopreventive effects of tea have been presented but others and we suggest that tea components target specific cell-signaling pathways responsible for regulating cellular proliferation or apoptosis. These pathways include signal transduction pathways leading to activator protein-1 (AP-1) and/or nuclear factor kappa B (NF-kappaB). AP-1 and NF-kappaB are transcription factors that are known to be extremely important in tumor promoterinduced cell transformation and tumor promotion, and both are influenced differentially by the MAP kinase pathways. The purpose of this brief review is to present recent research data from other and our laboratory focusing on the tea-induced cellular signal transduction events associated with the MAP kinase, AP-1, and NF-kappaB pathways. PMID- 12542979 TI - Chemoprevention of Helicobacter pylori-associated gastric carcinogenesis in a mouse model: is it possible? AB - Although debates still exist whether Helicobacter pylori infection is really class I carcinogen or not, H. pylori has been known to provoke precancerous lesions like gastric adenoma and chronic atrophic gastritis with intestinal metaplasia as well as gastric cancer. Chronic persistent, uncontrolled gastric inflammations are possible basis for ensuing gastric carcinogenesis and H. pylori infection increased COX-2 expressions, which might be the one of the mechanisms leading to gastric cancer. To know the implication of long-term treatment of antiinflammatory drugs, rebamipide or nimesulide, on H. pylori-associated gastric carcinogenesis, we infected C57BL/6 mice with H. pylori, especially after MNU administration to promote carcinogenesis and the effects of the long-term administration of rebamipide or nimesulide were evaluated. C57BL/6 mice were sacrificed 50 weeks after H. pylori infection. Colonization rates of H. pylori, degree of gastric inflammation and other pathological changes including atrophic gastritis and metaplasia, serum levels and mRNA transcripts of various mouse cytokines and chemokines, and NF-kappaB binding activities, and finally the presence of gastric adenocarcinoma were compared between H. pylori infected group (HP), and H. pylori infected group administered with long-term rebamipide containing pellet diets (HPR) or nimesulide mixed pellets (HPN). Gastric mucosal expressions of ICAM-1, HCAM, MMP, and transcriptional regulations of NF-kappaB binding were all significantly decreased in HPR group than in HP group. Multi probe RNase protection assay showed the significantly decreased mRNA levels of apoptosis related genes and various cytokines genes like IFN-gamma, RANTES, TNF alpha, TNFR p75, IL-1beta in HPR group. In the experiment designed to provoke gastric cancer through MNU treatment with H. pylori infection, the incidence of gastric carcinoma was not changed between HP and HPR group, but significantly decreased in HPN group, suggesting the chemoprevention of H. pylori-associated gastric carcinogenesis by COX-2 inhibition. Long-term administration of antiinflammatory drugs should be considered in the treatment of H. pylori since they showed the molecular and biologic advantages with possible chemopreventive effect against H. pylori-associated gastric carcinogenesis. If the final concrete proof showing the causal relationship between H. pylori infection and gastric carcinogenesis could be obtained, that will shed new light on chemoprevention of gastric cancer, that is, that gastric cancer could be prevented through either the eradication of H. pylori or lessening the inflammation provoked by H. pylori infection in high risk group. PMID- 12542980 TI - Oxidative stress, chromatin remodeling and gene transcription in inflammation and chronic lung diseases. AB - Inflammatory lung diseases are characterized by chronic inflammation and oxidant/antioxidant imbalance. The sources of the increased oxidative stress in patients with chronic inflammatory lung diseases such as asthma and chronic obstructive pulmonary disease (COPD) derive from the increased burden of inhaled oxidants, and from the increased amounts of reactive oxygen species (ROS) generated by several inflammatory, immune and various structural cells of the airways. Increased levels of ROS produced in the airways is reflected by increased markers of oxidative stress in the airspaces, sputum, breath, lungs and blood in patients with lung diseases. ROS, either directly or via the formation of lipid peroxidation products such as 4-hydroxy-2-nonenal may play a role in enhancing the inflammation through the activation of stress kinases (JNK, MAPK, p38) and redox sensitive transcription factors such as NF-capital KJE, MacedonianB and AP-1. Recent evidences have indicated that oxidative stress and pro-inflammatory mediators can alter nuclear histone acetylation/deacetylation allowing access for transcription factor DNA binding leading to enhanced pro inflammatory gene expression in various lung cells. Understanding of the mechanisms of redox signaling, NF-kappaB/AP-1 regulation, the balance between histone acetylation and deacetylation and the release and expression of pro- and antiinflammatory mediators may lead to the development of novel therapies based on the pharmacological manipulation of antioxidants in lung inflammation and injury. Antioxidants that have effective wide spectrum activity and good bioavailability, thiols or molecules which have dual antioxidant and anti inflammatory activity, may be potential therapeutic agents which not only protect against the direct injurious effects of oxidants, but may fundamentally alter the underlying inflammatory processes which play an important role in the pathogenesis of chronic inflammatory lung diseases. PMID- 12542981 TI - Histone deacetylase in carcinogenesis and its inhibitors as anti-cancer agents. AB - The acetylation state of histone is reversibly regulated by histone acetyltransferase (HAT) and deacetylase (HDAC). An imbalance of this reaction leads to an aberrant behavior of the cells in morphology, cell cycle, differentiation, and carcinogenesis. Recently, these key enzymes in the gene expression were cloned. They revealed a broad use of this modification, not only in histone, but also other proteins that involved transcription, nuclear transport, and cytoskeleton. These results suggest that HAT/HDAC takes charge of multiple-functions in the cell, not just the gene expression. HDAC is especially known to play an important role in carcinogenesis. The enzyme has been considered a target molecule for cancer therapy. The inhibition of HDAC activity by a specific inhibitor induces growth arrest, differentiation, and apoptosis of transformed or several cancer cells. Some of these inhibitors are in a clinical trial at phase I or phase II. The discovery and development of specific HDAC inhibitors are helpful for cancer therapy, and decipher the molecular mode of action for HDAC. PMID- 12542982 TI - Hypoxia-induced angiogenesis during carcinogenesis. AB - The formation of new blood vessels, angiogenesis, is an essential process during development and disease. Angiogenesis is well known as a crucial step in tumor growth and progression. Angiogenesis is induced by hypoxic conditions and regulated by the hypoxia-inducible factor 1 (HIF-1). The expression of HIF-1 correlates with hypoxia-induced angiogenesis as a result of the induction of the major HIF-1 target gene, vascular endothelial cell growth factor (VEGF). In this review, a brief overview of the mechanism of angiogenesis is discussed, focusing on the regulatory processes of the HIF-1 transcription factor. HIF-1 consists of a constitutively expressed HIF-1 beta (HIF-1beta) subunit and an oxygen-regulated HIF-1 alpha (HIF-1a) subunit. The stability and activity of HIF-1alpha are regulated by the interaction with various proteins, such as pVHL, p53, and p300/CBP as well as by post-translational modifications, hydroxylation, acetylation, and phosphorylation. It was recently reported that HIF-1alpha binds a co-activator of the AP-1 transcription factor, Jab-1, which inhibits the p53 dependent degradation of HIF-1 and enhances the transcriptional activity of HIF-1 and the subsequent VEGF expression under hypoxic conditions. ARD1 acetylates HIF 1alpha and stimulates pVHL-mediated ubiquitination of HIF-1alpha. With a growing knowledge of the molecular mechanisms in this field, novel strategies to prevent tumor angiogenesis can be developed, and from these, new anticancer therapies may arise. PMID- 12542983 TI - Roles of matrix metalloproteinases in tumor metastasis and angiogenesis. AB - Matrix metalloproteinases (MMPs), zinc dependent proteolytic enzymes, cleave extracellular matrix (ECM: collagen, laminin, firbronectin, etc) as well as non matrix substrates (growth factors, cell surface receptors, etc). The deregulation of MMPs is involved in many diseases, such as tumor metastasis, rheumatoid arthritis, and periodontal disease. Metastasis is the major cause of death among cancer patients. In this review, we will focus on the roles of MMPs in tumor metastasis. The process of metastasis involves a cascade of linked, sequential steps that involve multiple host-tumor interactions. Specifically, MMPs are involved in many steps of tumor metastasis. These include tumor invasion, migration, host immune escape, extravasation, angiogenesis, and tumor growth. Therefore, without MMPs, the tumor cell cannot perform successful metastasis. The activities of MMPs are tightly regulated at the gene transcription levels, zymogen activation by proteolysis, and inhibition of active forms by endogenous inhibitors, tissue inhibitor of metalloproteinase (TIMP), and RECK. The detailed regulations of MMPs are described in this review. PMID- 12542984 TI - Molecular aspects of hepatitis B viral infection and the viral carcinogenesis. AB - Of many viral causes of human cancer, few are of greater global importance than the hepatitis B virus (HBV). Over 250 million people worldwide are persistently infected with HBV. A significant minority of these develop severe pathologic consequences, including chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC). Earlier epidemiological evidence suggested a link between chronic HBV infection and HCC. Further, the existence of related animal viruses that induce acute and chronic infections of the liver, and eventually HCC, confirms the concept that HBV belongs to one of the few human oncogenic viruses. Although it is clear that chronic HBV infections are major risk factors, relatively little is understood about how the viral factors contribute to hepatocarcinogenesis. This review will introduce molecular aspects of the viral infection, and highlight recent findings on the viral contribution to hepatocarcinogenesis. PMID- 12542985 TI - Radiation-induced tumorigenesis. AB - During the past 2 decades, radiation tumorigenesis researchers have focused on cellular and molecular mechanisms. We reviewed some of these research fields, since they may specifically relate to the induction of cancer by ionizing radiation. First, radiation-mediated mutation was discussed. Then the initiating event in radiation carcinogenesis, as well as other genetic events that may be involved, is discussed in terms of the possible role of the activation of genes and the loss of cell-cycle checkpoints. PMID- 12542986 TI - Lessons from drug allergy: against dogmata. PMID- 12542987 TI - The role of epinephrine in the treatment of anaphylaxis. AB - Epinephrine is the cornerstone of anaphylaxis management. Its administration should be immediate upon evidence of the occurrence of anaphylaxis. Delays in administration may be fatal. The most appropriate administration is 0.3 to 0.5 mL of 1:1000 dilution intramuscularly for adults and 0.01 mg/kg for children, given in the lateral thigh. Patients with known anaphylactic reactivity should be prescribed an epinephrine auto-injector to be carried at all times for treatment of potential recurrences. Education of the patient or parent regarding the proper use of this tool is paramount. PMID- 12542988 TI - Exercise-induced Anaphylaxis. AB - Exercise-induced anaphylaxis has been recognized with increasing frequency since its original description in 1980. Recent studies suggest food-induced reactions may occur frequently in this syndrome, which is a mast cell-dependent phenomenon. In this article, the clinical manifestations of exercise-induced anaphylaxis are reviewed, and food-related factors contributing to the disorder are considered. PMID- 12542989 TI - Immunopharmacology of hypersensitivity reactions to drugs. AB - Drug hypersensitivity reactions are characterized by their unpredictability, lack of simple dose-dependency, host sensitivity, and potentially serious clinical outcome. They occur in a small proportion of patients, and usually the predisposing factors are unknown, although there is increasing evidence for genetic predisposition and disease being significant risk factors. The current understanding of the chemical basis of immune-mediated reactions is based on the hapten hypothesis, which requires drug bioactivation, covalent binding to proteins, followed by uptake, antigen processing, and a polyclonal immune response. The recently proposed "danger hypothesis" can be considered to be an essential addition to the hapten hypothesis. According to the danger hypothesis, the immune response to a drug-derived antigen requires the presence of co stimulatory signals and cytokines, which propagate and determine the type of immune response. The "danger signal" might result from chemical, physical, or viral stress. PMID- 12542990 TI - Epidemiology of anaphylaxis in the United States. AB - Anaphylaxis is a potentially life-threatening allergic reaction characterized by a syndrome of dermatologic and systemic signs and symptoms, at times including cardiovascular collapse from distributive shock. Although it is recognized that the incidence of anaphylaxis is increasing, the extent of its prevalence, both from individual causes and in toto, remains unclear. Common causes include pharmacologic agents, such as antibiotics and radiocontrast media, as well as foods, insect stings, and latex exposure. The available literature suffers from methodologic shortcomings, limiting the accuracy of estimated prevalence. However, current data suggest that there are approximately 1500 annual deaths from anaphylaxis, and between 2.8 million and 42.7 million Americans are at risk of experiencing an episode of anaphylaxis during their lives. PMID- 12542993 TI - Psychological aspects of food allergy. AB - Food allergies may impact the emotions of patients through both direct and indirect mechanisms. Direct mechanisms include the effects on the central nervous system from biologic mediators released during an allergic reaction to food. Indirect mechanisms include the stress of coping with a food allergy--for example, food preparation and avoidance--as well as managing the fear of the potential consequences of ingesting the food. Indirect effects may also be mediated through family members--for example, the impact of a parent's stress on the child. These relationships are difficult to study, in part because many patients who report food allergy symptoms do not have objective symptoms when challenged with the offending food. Symptoms may be misinterpreted as food allergy more often by patients with certain psychological profiles. In this paper, relevant literature is reviewed, and clinical treatment designed to minimize the emotional suffering of patients and their families is presented through the description of a case vignette. PMID- 12542994 TI - Latex-fruit syndrome. AB - Natural rubber latex immunoglobulin E-mediated hypersensitivity is probably one of the most relevant challenges that has been faced in the treatment of allergies during recent years. Additionally, allergen cross-reactivity has arisen as another very important problem, in the difficulty in diagnosing it and in its clinical implications. It is clear that some latex allergens cross-react with plant-derived food allergens, the so-called latex-fruit syndrome, with evident clinical consequences. Although the foods most frequently involved are banana, avocado, kiwi, and chestnut, several others are also implicated. Investigations point to a group of defense-related plant proteins, class I chitinases, which cross-react with a major latex allergen, hevein, as the panallergens responsible for the syndrome. This review focuses on our current understanding of the latex fruit syndrome. PMID- 12542995 TI - Tree nut allergy. AB - Tree nuts are clinically associated with severe immunoglobulin E-mediated systemic allergic reactions independent of pollen allergy and with reactions that are usually confined to the oral mucosa in patients with immunoglobulin E directed toward cross-reacting pollen allergens. The latter reactions can progress to severe and life-threatening episodes in some patients. Many patients with severe tree nut allergy are co-sensitized to peanut. Clinical studies on cross-reactivity between the tree nuts are few in number, but based on reports to date, avoidance of the other tree nuts once sensitivity is diagnosed appears prudent unless specific challenges are performed to ensure clinical tolerance. Even then, great care must be taken to avoid cross-contamination. As with other severe food allergies, a recurrent problem in clinical management is the failure of physicians to prescribe self-injectable epinephrine to patients who are at risk of anaphylaxis. PMID- 12542996 TI - Adverse reactions to food additives. AB - There are thousands of additives used by the food industry for a variety of purposes in the foods we eat. However, only a small number have been implicated in causing adverse reactions in humans. Although there are reported cases of individuals who have reactions to single additives, most of the medical literature involves patients with asthma or chronic idiopathic urticaria/angioedema whose conditions are exacerbated after ingestion of food additives. Many of these reports are characterized by poorly controlled challenge procedures. Recent studies performed under properly controlled conditions imply that sensitivity to food additives in patients with chronic urticaria/angioedema is very uncommon. PMID- 12542998 TI - Animal models of allergic asthma. AB - We are fortunate to have many species of animals that can serve as adequate models for allergic disease in humans. This review is focused mostly on models of allergic airway disease, and some major categories of animals used for asthma research are discussed, including rodents and nonhuman primates. Furthermore, evidence that supports and criticizes the use of animal models of asthma is provided. There is no animal model that exactly reproduces the pathology of human asthma. However, these models are necessary for the development of novel therapies and an understanding of the detailed pathogenesis of the response of mammals to respirable allergens. PMID- 12542999 TI - Eosinophilic gastroenteritis: a problem of the mucosal immune system? AB - The gastrointestinal immune system is a major component of the mucosal barrier, which provides an appropriate immunologic homeostasis between host and numerous foreign antigens, including microbial and dietary antigens. However, under certain pathological circumstances created by disturbance of the immunologic balance, allergic responses associated with the gastrointestinal tract can be triggered by abnormal immune responses against selected food protein antigens. Among the several types of immune competent cells, eosinophils are generally considered to play a central role for the development of allergic diseases in gastrointestinal tissue. Although most research has been focused on the molecular and cellular understanding of eosinophils in the peripheral tissues and lung, recent studies elucidate the unique trafficking and regulation mechanisms of eosinophils in the gastrointestinal tissues. In this review, we summarize current findings in the regulatory mechanism of gastrointestinal eosinophils. Furthermore, several unique murine models for eosinophilic gastroenteritis, which can be applied for the elucidation of underlying mechanisms of eosinophil mediated gastrointestinal allergy, and the development of new mucosal immune therapy for the control of food allergy are reviewed. PMID- 12543000 TI - Experimental autoimmune encephalomyelitis: cytokines, effector T cells, and antigen-presenting cells in a prototypical Th1-mediated autoimmune disease. AB - Experimental autoimmune encephalomyelitis (EAE) is widely depicted as the prototypical CD4+ Th1-mediated autoimmune disease. Microglia and perivascular macrophages are believed to act as antigen-presenting cells during the effector phase of EAE. In this article, recent data that challenge these conceptions are reviewed. Several recent studies have shown that myelin-reactive CD8+ T cells can mediate inflammatory demyelination. Furthermore, dendritic-like cells have been detected in EAE lesions and implicated in encephalitogenic T-cell activation. Although Th1 polarizing monokines, such as interleukin-12 (IL-12) and possibly IL 23, are critical for the manifestation of EAE, individual Th1 effector cytokines were found to be dispensible. PMID- 12543039 TI - Macrophages and post-burn immune dysfunction. AB - The activation of a pro-inflammatory cascade after burn injury appears to be important in the development of subsequent immune dysfunction, susceptibility to sepsis and multiple organ failure. Macrophages are major producers of pro inflammatory mediators and their productive capacity for these mediators is markedly enhanced following thermal injury. Thus, macrophage hyperactivity (as defined by increased productive capacity for pro-inflammatory mediators) appears to be of critical importance in the development of post-burn immune dysfunction. This review will focus on the current state of knowledge with regards to the role of macrophages in the development of post-burn immune dysfunction. Particular areas of discussion include: nitric oxide synthase (NOS) and cyclooxygenase (COX) enzyme systems, macrophages and the T-helper (Th)-1/Th-2 cytokine responses, alterations in macrophages signal transduction and a potential role for gamma/delta T-cells in the development of macrophage hyperactivity following thermal injury. A more comprehensive understanding of the relationship between macrophage activity and post-burn immune dysfunction will hopefully provide the basis for improved therapeutic regimes in the treatment of burn patients. PMID- 12543040 TI - What's new in burn microbiology? James Laing Memorial Prize Essay 2000. AB - A variety of factors contribute to the development of infection in burned patients. The role of wound management procedures, risk factors associated with infection, typical bacterial pathogens and associated exotoxins, current problems with antibiotic resistance, wound sampling and rare complications of infection are described. The use of new novel treatments that are currently being developed, such as cell signalling molecules and the increasing use of natural antimicrobial agents, for example honey, papaya fruit and tea-tree oil are discussed. The impact of new methods for earlier detection of infectious agents that could change future practices in burn care is also described. PMID- 12543041 TI - Social competence and behaviour problems in burned children. AB - The aim of this study was to collect follow-up data on social competence and behavioural problems in a sample of Swedish burned children and to compare the results with normative data from a reference group of children comparable in age, socio-economic status and gender. Parents of 44 children (55% response rate) aged 7-12 years were asked to complete a questionnaire booklet including the Children's Behaviour Questionnaire (CBQ) and the Social Competence Inventory (SCI). Data from the children's teachers were also collected for 20 children using the same booklet. In addition, data on TBSA, localisation of injury, and other background factors were collected. Results showed that the burned children were rated by their parents as showing lesser degrees of social initiative and more externalising problems and concentration problems compared with the control group. Teachers rated the burn injured children as having less prosocial orientation, more externalising problems, and more concentration problems. No clear effects were found for gender and characteristics of the burn injury. Results on the Social Competence Inventory were associated with scores on the Children's Behaviour Questionnaire.The findings are consistent with previous research in that the differences found were relatively small. However, they do call for attention to the possible adverse effects of growing up with a burn injury, but also to the possible pre-morbid characteristics that may be related to the injury. PMID- 12543042 TI - Burnt wives--a study of suicides. AB - Ritual self-immolation has long been practiced in India. Although in the past the practice has been related to the act of sati, it is currently more commonly associated with dowry disputes. In India, dowries are a continuing series of gifts endowed before and after the marriage. When dowry expectations are not met, the young bride may be killed or compelled to commit suicide, most frequently by burning. In a cohort of 152 burned wives, 32 (21%) were immolation suicides and these cases were analyzed from both epidemiological and medicolegal aspects. Suicide by burning amongst women is a major concern in India as it has become pervasive throughout all social strata and geographical areas. In this series, most women were from joint families (i.e. multigenerational groups of related individuals living under a single roof) and the suicides occurred 2-5 years after marriage. The majority of the affected wives were 21-25 years of age (69%) at the time of the suicide and sustained more than a 50% TBSA burn injury after dousing themselves with kerosene. Most died at the time of the incident or within the subsequent 24h, most commonly from shock. PMID- 12543043 TI - Therapeutic effects of IL-12 combined with benzoylmesaconine, a non-toxic aconitine-hydrolysate, against herpes simplex virus type 1 infection in mice following thermal injury. AB - IL-12 is an inducer of type 1 T cell responses, which are essential in host defense against herpes simplex virus type 1 (HSV-1) infection. However, type 1 T cell responses are not elicited by IL-12 in thermally injured mice (TI-mice) that routinely have a predominance of burn-associated type 2 T cell responses. In our previous studies, benzoylmesaconine (BEN, an aconitine derivative extracted from heated-Aconiti tuber) induced the generation of CD4+ T cells antagonistic to type 2 T cells (BEN-CD4+ T cells). In the present study, the effects of a combination therapy using IL-12 and BEN to treat severe HSV-1 infection in TI-mice were investigated. When TI-mice were treated with either IL-12 (500 U per mouse) or BEN (1 microg/kg) alone, they did not resist HSV-1 infection. However, 60-80% of TI-mice exposed to HSV-1 survived after they received IL-12 and BEN or BEN-CD4+ T cells in combination. After stimulation with anti-CD3 mAb in vitro, IFN- was not produced in cultures of splenic T cells from TI-mice exposed to HSV-1 and treated with either IL-12, BEN or BEN-CD4+ T cell alone. However, IFN- production was induced by the mAb stimulation in the cultures of T cells from infected mice treated with IL-12 and BEN or BEN-CD4+ T cells in combination. These results suggest that the combination therapy of IL-12 (an inducer of type 1 T cell responses) and BEN (an inhibitor of type 2 T cell responses) may protect TI-mice from severe HSV-1 infection. PMID- 12543044 TI - Effects of dietary arginine supplementation on antibody production and antioxidant enzyme activity in burned mice. AB - This study investigated the effect of arginine (Arg) supplementation on specific antibody production and antioxidant enzyme activities in burned mice vaccinated with detoxified Pseudomonas exotoxin A linked with the outer membrane proteins I and F, named PEIF. Also, the survival rate of burned mice complicated with Pseudomonas aeruginosa was evaluated. Experiment 1: Thirty BALB/c mice were assigned to two groups. One group was fed a control diet with casein as the protein source, while the other group was supplemented with 2% Arg in addition to casein. The two groups were isonitrogenous. The mice were immunized twice with PEIF, and the production of specific antibodies against PEIF was measured every week. After 8 weeks, all mice received a 30% body surface area burn injury. Mice were sacrificed 24h after the burn. The antioxidant enzyme activities and lipid peroxides in the tissues as well as the specific antibody production were analyzed. Experiment 2: Twenty-eight mice were divided into two groups and vaccinated as described in experiment 1. After the burn the mice were infected with P. aeruginosa, and the survival rate was observed for 8 days. The results demonstrated that antioxidant enzyme activities and lipid peroxides in tissues were significantly lower in the Arg group than in the control group after the burn. The production of specific antibodies against P. aeruginosa significantly increased in the Arg group at 4 and 7 weeks after immunization, and 24h after the burn. The survival rates of vaccinated burned mice after bacterial infection did not significantly differ between the two groups. These results suggest that vaccinating mice with Arg supplementation may enhance humoral immunity and attenuate the oxidative stress induced by burn injury. However, Arg supplementation did not improve survival in vaccinated mice complicated with P. aeruginosa infection. PMID- 12543045 TI - Treatment of patients with severe burn injuries: the impact of schizophrenia. AB - Four patients treated in one ward of a psychiatric clinic were admitted to our burn unit within 2 months due to severe burn injuries. The patients showed signs of a self-mutilation epidemic. All four patients were female and the mean age was 28 years. The psychiatric diagnosis was schizophrenia in all patients (ICD 10: F20.9). The ignition of flammable liquid was the most common method and the mean burned TBSA was 33%. The mean severity score (ABSI) was 8 and the median hospital stay was 50 days. All patients were characterised by a prolonged hospital stay in comparison to patients without additional psychiatric pathology (median 31 days). This prolonged stay was based on a delayed wound healing, more operations, extended time for mobilisation and difficulties in co-operation. It is possible that in patients with schizophrenia, changes in nutrition, activity, sleep and drug use could influence their immune system profoundly. Anxiety and depression is also associated with the impairment of cellular and humoural immunity. Poor sleep reduces the production of an anabolic endocrine environment and sleep disturbances can interfere with macrophage and lymphocyte functions. Poor appetite leads to malnutrition, which is also capable of producing delayed wound healing. On the other hand, apathy and a general lack of motivation interfere with therapeutic strategies, because poor appetite and weight loss often occurs after neuroleptic withdrawal, which is correlated with clinical decompensation. Moreover, this "self-destructive" behaviour, which is acting on the immune system, might make a patient more susceptible to infection. All these aspects and side effects of schizophrenia combine to make the treatment of burned patients with schizophrenia a very special and difficult task. PMID- 12543046 TI - The prevalence of HIV infection among burn patients in a burns unit in Malawi and its influence on outcome. AB - In a 1 year study, 342 patients admitted to the Burns Unit at Queen Elizabeth Central Hospital (QECH), Blantyre, Malawi underwent a voluntary HIV test. Forty (11.7%) tested HIV positive: of those aged above 15 years, 31% (34 out of 112) were HIV positive, whilst 3% (6 of 231) aged under 15 were HIV positive of whom the majority were in those aged under 5 years (5 of 125, 4%). Patients who were HIV positive had an increased risk of death (P=0.04) which was mainly due to sepsis, but those HIV patients, who did not develop infection or recovered from an episode of sepsis, had similar hospital stay, need for skin grafting and graft take as nonHIV patients. There was no difference in pathogens cultured from wound swabs taken from HIV positive and negative patients. HIV positive patients had significantly lower CD4 counts as compared to HIV negative patients (mean 383mm3 (S.D. 320) and 937mm3 (S.D. 497), respectively). However, low CD4 counts were also found in the HIV negative patients (mean 901, range 131-1964) and 24% had CD4 <500/mm3. Both HIV status and the total body surface area (TBSA) burned were independent predictors of CD4 count. TBSA was an independent risk factor for death (odds ratio 1.3; 95% CI 1.1, 1.4). In patients with TBSA burns of over 30%, mortality approached 100% irrespective of HIV status, but in patients with burns of 11-20% TBSA and who were HIV positive have a mortality of 25% compared to 12% in HIV negative patients; for 21-30% TBSA burns mortality was 100% compared to 50% for HIV positive and HIV negative patients, respectively. PMID- 12543047 TI - Shabbes burn, a burn that occurs solely among Jewish orthodox children; due to accidental shower from overhead water heaters. AB - From January 1990 to January 2000, 35 children were treated in our department for Shabbes burn, a unique scald burn that occurred mainly among orthodox Jews during the Sabbath. A retrospective review was conducted to determine the extent of the problem, to understand the burn mechanism and to suggest a prevention program. A shower of hot water from the Sabbath heater is the cause of this burn. Among the Shabbes burn cases, 27 patients were female (77%) and 17 children (48%) were between 3 and 6 years old. It is suggested that this is a common burn that occurs among orthodox Jewish families and affects mainly females. Education programs using the media directed to the Jewish orthodox population have been conducted, this combined with redesigned of the heater, have reduced significantly the incidence of the burn as seen in our institute. However, even though efforts have been supported widely, there remains a need for educational and governmental regulations on a national level. This could aid orthodox Jews not only in Israel but globally, as well. PMID- 12543048 TI - Experience of 14 years of emergency reconstruction of electrical injuries. AB - Although there have been great advances in the treatment of electrical injuries in the last 20 years, the extremity loss ratio in electrical injuries remains at an unacceptably high level. The primary cause is due to the progressive tissue necrosis which results in the continuous extension of necrosis in the wound, leading to loss of the whole injured extremity. This study reports attempts to break the dangerous tissue necrosis circle and save the form and function of damaged extremities. After 14 years of systematic experimental and clinical studies a successful comprehensive urgent reconstruction alternative (CURA) for electrical injuries is proposed. CURA includes: debriding the wound as early as possible after injury; preserving the vital tissue structures as much as possible, such as nerves, vessels, joints, tendons, bone, even though they have undergone devitalization or local necrosis; repairing these vital tissues during the first surgery if functional reconstruction requires it; protecting the wound bed by covering with tissue flaps of rich blood supply; improving flap survival through moist dressings supported by continuous irrigation beneath the flaps for a 24-72h period after surgery with measures to control local infection; and last, giving general systemic treatment with vasoactive agents and antibiotics. Four hundred and fifty nine wounds in 155 patients suffering from electrical injuries have been successfully treated with this technique between 1986 and 2000 and are reported in this paper. Satisfactory results were obtained with the extremity loss proportion reduced to less than 9% compared with 41.5% during the 10 years before 1984 in the same hospital. The authors suggest that CURA is an effective and workable method for treatment of electrical injuries. PMID- 12543049 TI - Radiological manifestation of pulmonary complications in deceased intensive care burn patients. AB - The chest radiographs of 46 burn patients who died in the burn intensive care unit (BICU) were retrospectively analyzed to study the spectrum of pulmonary complications and their contribution to patient's mortality. There were 25 male and 21 female patients and their mean ages were 34 and 30 years, respectively. Forty-three patients had flame burns, two chemical, and one scald with a mean total burn surface area (TBSA) of 71%. Thirty-six of them had inhalation injury and of these 25 patients developed septicaemia. Out of these 46 patients, 39 had a total of 60 pulmonary complications on various postburn days. The commonest complications were consolidation (28.3%) and adult respiratory distress syndrome (ARDS) (26.7%) mainly due to inhalation injury and/or following septicaemia. The majority of these complications (46.7%) occurred in the late phase (postburn day 5 onwards). Forty-one (89.2%) patients died due to multi-organ failure (MOF) and a good number of them had secondary respiratory failure. The flame burn patients with large TBSA, presence of inhalation injury, and occurrence of septicaemia, are at risk for pulmonary complications that equally affect adult males and females. Pulmonary complications irrespective of the cause significantly contribute to the mortality. This study suggests that serial chest X-rays done in BICU form an important diagnostic tool for pulmonary complications from postburn day 1 onwards, and is useful for subsequent monitoring of the treatment. All burn intensive care units may not be privileged to have a full time radiologist, and intensivist. Therefore, the burn surgeon needs to metamorphose into an intensivist and double as a burn radiologist for early detection and quick treatment if his surgical skills are to be adequately rewarded. PMID- 12543051 TI - Unusual inflicted hot oil burns in a 7-year-old. AB - Pediatric burn injuries occur not infrequently as the result of abuse. While the majority of these burns are inflicted scald burns, those due to contact or contact/scald mechanisms may present diagnostic challenges. A child with unusual combined contact and scald burns caused by a metal spatula heated in hot cooking oil is described. The odd pattern of healed injury limited the initial diagnosis to inflicted healing burns. Despite the initial lack of disclosure, an inflicted etiology was supported by clearly delineated margins, macular lesions with hyper pigmented rims and variegated central regions. Additional findings of numerous adult bites and bruises provided adjunctive support for an inflicted etiology. Aspects of this case are atypical for the usual demographics of a burn victim. This case extends the known presentations of inflicted contact/scald burns. PMID- 12543050 TI - Effect of diagnosis and treatment of sinusitis in critically ill burn victims. AB - The purpose of this study was to assess the incidence of diagnosed sinusitis and the effect of diagnosis and treatment on the outcome in critically ill burn victims. Chart analysis of 84 consecutive burn victims requiring mechanical ventilation for greater than 7 days was performed. Sinusitis was diagnosed in 13/84 patients (15%). There was no difference in age or total body surface area burned, or the incidence of inhalation injury, ARDS, pneumonia and sepsis (P>0.05). Co-morbid disease was similar in both the groups. The number of ventilator-dependent days and hospital length of stay were higher in the sinusitis group (P<0.05). The hospital mortality in those diagnosed and treated for sinusitis was 23% (3/13) as opposed to 48% (34/71) in those not diagnosed with sinusitis (P<0.05). Increased number of ventilator-dependent days and longer hospital stay are associated with the diagnosis of sinusitis. Our findings suggest an improved survival in those diagnosed and treated for sinusitis. PMID- 12543052 TI - Bilateral free myocutaneous latissimus dorsi flap repair of the upper limb amputation stumps due to electrical injury. PMID- 12543053 TI - Finger ischemia secondary to the synergistic agonist effect of norepinephrine and ergonovine and in a burn patient. PMID- 12543054 TI - Infant scald burns: a case of negligence? PMID- 12543055 TI - Maggot debridement therapy. PMID- 12543056 TI - The long-term consequences of lightning injuries. Muehlberger T, Vogt PM, Munster AM, (Burns 2001;27:829-33). PMID- 12543057 TI - Current St John's wort research from mode of action to clinical efficacy. AB - Preparations from St. John's wort extracts are used in the treatment of depression in many countries and represent an accepted alternative to synthetic antidepressants or behavioural therapy. St. John's wort extracts are therefore used in a therapeutic area which extends well beyond the traditional field of herbal medicine. The current status of preclinical and clinical research is summarised. St. John's wort extract has a clear inhibitory effect on the neuronal uptake not only of serotonin, noradrenaline, and dopamine but also of gamma aminobutyric acid (GABA) and L-glutamate. No other antidepressant shows an approximately equally broad inhibitory profile. In good agreement with the effects in various biochemical models of antidepressant action, many effects in a number of behavioural pharmacology models of antidepressant efficacy could also be demonstrated for St. John's wort extract. Similar doses of John's wort also cause changes in the above-mentioned neurotransmitter systems in the brain. Out of all individual substances of St. John's wort only hyperforin and its structural analogue adhyperforin inhibit the re-uptake of the investigated neurotransmitters. However, hyperforin does not act as a competitive inhibitor at the transmitter binding sites of the transporter proteins but it affects the sodium gradient which then leads to an inhibition of uptake. The broad spectrum of action which characterises St. John's wort extracts has only been described for the pure substance hyperforin. Over the past year a number of good clinical studies have been carried out which confirm the efficacy and tolerability of St. John's wort extracts in mild depressive disorders, even if the therapeutic efficacy has recently been questioned by an American study. All studies have confirmed the good tolerability of St. John's wort extract and the very low frequency of adverse events. However, some drug interactions have been found to occur with St. John's wort extract, a number of which are of clinical relevance. In summary, pharmacological activity and therapeutic efficacy of St. John's wort extract as an antidepressant are supported by a large number of scientific publications. Within the wide range of components in St. John's wort extract, hyperforin plays an important, if not an outstanding role. PMID- 12543058 TI - Anthracycline-induced cardiac toxicity is not increased by dietary omega-3 fatty acids. AB - Exogenous n-3 polyunsaturated fatty acids (PUFA) and specially docosahexaenoic acid (DHA) have been previously reported to potentiate the efficacy of anticancer agents that generate an oxidative stress, such as anthracyclines, by enhancing the susceptibility of cell membranes to lipid peroxidation. Since lipid peroxidation has also been suggested to mediate anthracycline-induced heart failure, we designed a study aimed at investigating whether a DHA-enriched diet coupled with controlled oxidative conditions prevents or aggravates this serious side effect in vivo. Female Sprague-Dawley rats were submitted for at least 3 weeks to diet enriched in DHA, which was provided either as natural oil (sardine oil, experiment 1) or in a purified form (DHASCO, experiment 2). At the same time, to constrain the nutritional oxidative status, the anti-oxidant Vitamin E or the pro-oxidant menadione/sodium ascorbate redox mixture was added. Then, epirubicin was administered weekly at two cumulative doses, 9 mg x kg(-1) (experiment 1) or 15 mg x kg(-1) (experiment 2). Cardiotoxicity was assessed by electrocardiographic (ECG) and hemodynamic measurements, completed with histological examination. Epirubicin-induced dose-dependent mortality, alterations of hemodynamic parameters and histological damages, all features characterizing the occurrence of congestive heart failure. Moreover, the addition of anti- or pro-oxidant did not change the hemodynamics either at the lowest (experiment 1) or the highest dose (experiment 2). Similarly, the ECG measurements and histological examinations did not reveal any difference. DHA was actually incorporated, as evaluated through the adipose tissue fatty acid composition. All these observations indicated that the DHA-enriched diet, placed under controlled oxidative conditions, did not appear to prevent but neither to aggravate epirubicin-induced cardiotoxicity. These findings support the idea that DHA improves the anthracycline therapeutic index. PMID- 12543059 TI - Pravastatin inhibits pro-inflammatory effects of Alzheimer's peptide Abeta(1-42) in glioma cell culture in vitro. AB - Statins are known to exert a number of biological effects apart from reducing cholesterol synthesis. The results of recent studies indicate that patients treated with pravastatin have a lower prevalence of diagnosed Alzheimer's disease (AD). These observations prompted us to examine the effects of pravastatin on Alzheimer's peptide (Abeta(1-42))-induced pro-inflammatory activation in the human glioma cell line in vitro. Cells alone or cells pre-treated with pravastatin (0.1mg x ml(-1)) for 24h were stimulated with 5 microM of freshly dissolved Abeta(1-42) for the next 24h. The pre-treatment of cells with pravastatin diminished the capacity of Abeta to induce metalloproteinases, cytokine IL-6 and free radical levels. Although both pravastatin and Abeta(1-42) separately increased PPARgamma activity, the combination of Abeta(1-42) and pravastatin resulted in no effect on PPARgamma expression. These data indicate that soluble forms of Abeta(1-42), which are a potent stimulus of pro inflammatory activation of glioma cells in vitro, could be a good target for pravastatin. PMID- 12543060 TI - The influence of midazolam and flumazenil on rat brain slices oxygen consumption. AB - This study investigated the impact of benzodiazepine receptor agonist, midazolam and antagonist, flumazenil, on the rat frontal cortex slices oxygen consumption (QO(2)), in presence and absence of gamma-aminobutyric acid (GABA). QO(2) was polarographically determined, using the biological oxygen monitor. As it was previously shown, GABA on its own decreases QO(2) moderately. Midazolam decreased QO(2) at 1.0mg/kg, whereas flumazenil had no effect. In combination with per se ineffective GABA (10(-6)mol/l), flumazenil showed respiratory depressant action, presumably revealing partial agonistic activity at some of GABA(A) receptor subtypes. However, it completely antagonized effects of midazolam on QO(2), on its own and in presence of GABA. Our results show that in vivo well-established effects of midazolam on cerebral metabolic activity could be reproduced in in vitro settings. Moreover, flumazenil antagonized this action, indicating the role of GABA(A)-benzodiazepine receptor complex activation in QO(2) regulation. PMID- 12543061 TI - Anticarcinogenic effect of bis-1,7-(2-hydroxyphenyl)-hepta-1,6-diene-3,5-dione a curcumin analog on DMH-induced colon cancer model. AB - 1,2-Dimethylhydrazine (DMH) is a toxic environmental pollutant which was reported also to be a colon-specific carcinogen. This study was performed to study the effect of bis-1,7-(2-hydroxyphenyl)-hepta-1,6-diene-3,5-dione, a bisdemethoxycurcumin analog (BDMC-A) on DMH-induced colon carcinogenesis in male Wistar rats and effects were compared with that of the reference drug, curcumin. Rats were given a weekly subcutaneous injection of DMH (20mg/kg body weight) in the groin, for 15 weeks. After a total experimental period of 32 weeks (including 2 weeks of acclimatization) tumor incidence was 100% in DMH-treated rats. Tumor was identified histologically as adenocarcinoma. Dysplasia, papillary pattern, cellular pleomorphism and carcinomatous glands were also noticed in DMH-treated rats. However, there was no colonic tumor in DMH+BDMC-A- and DMH+curcumin-treated rats but, lymphocyte infiltrations were observed. The levels of total bile acids and cholesterol in 24h fecal samples were significantly lower in DMH administered rats when compared to control rats, while, the excretion of bile acids and cholesterol were significantly increased and was near normal levels in DMH+BDMC-A and DMH+curcumin-treated rats. In DMH-induced tumor bearing rats the levels of colonic and intestinal cholesterol was significantly increased whereas, the levels of phospholipid was decreased with a concomitant increase in the activities of phospholipase A (PLA) and phospholipase C (PLC), compared to untreated control rats. Intragastric administration of BDMC-A and curcumin to DMH administered rats significantly lowered the cholesterol content and raised the phospholipid content and lowered the activities of PLA and PLC towards near normal values. Our study shows that the protective effect of BDMC-A during DMH induced colon carcinogenesis may be due to its modulatory effects on (i). histological changes, (ii). bile acids, (iii). cholesterol, and (iv). phospholipid metabolism in the target organ. Absence of histological changes in the colon of rats treated with BDMC-A, shows that long term administration of BDMC-A is nontoxic to experimental animals. Our study suggest that BDMC-A may emerge as a potent anticarcinogenic agent against colon cancer. As both BDMC-A and curcumin are equipotent in inhibiting the DMH-induced colon tumor incidence and normalizing histological changes, it could be concluded that the terminal phenolic group and the conjugated double bonds in the central seven carbon change may be responsible for the beneficial effects. PMID- 12543062 TI - Chemoprevention of DMBA-induced UV-B promoted, NOR-1-induced TPA promoted skin carcinogenesis, and DEN-induced phenobarbital promoted liver tumors in mice by extract of beetroot. AB - Our previous studies identified the extract of Beta vulgaris (beetroot), commercially also known as betanin, as a potent cancer chemopreventive agent in both in vitro Epstein-Barr early antigen activation assay and in an in vivo two stage mouse lung and skin carcinogenesis. To explore this issue further, we have now investigated its cancer chemopreventive potentials in three different chemical carcinogen initiation-promotion experimental tumor models in mice. Following tumor initiation with 390 nmol of 7,12-dimethylbenz(a)anthracene (DMBA) in 100 microl of acetone, the mouse skin tumor promotion with 3430 J/m(2) of ultraviolet light-B (UV-B) as well as splenomegaly was significantly inhibited by oral administration of 0.0025% betanin. At the same dose, betanin also afforded significant protection in the mouse skin cancer model following the topical application of 390 nmol of (+/-)-(E)-4-methyl-2-[(E)-hydroxyamino]-5-nitro-6 methoxy-3-hexanamide (NOR-1) in 100 microl of acetone and promoted by topical administration of 1.7 nmol of 12-O-tetradecanoylphorbol-13-acetate (TPA). In the two-stage model of hepatocarcinogenesis in mice with N-nitrosodiethylamine (DEN, 30 mg/kg) as the initiator and phenobarbital as the promoter, oral administration of 0.0025% betanin also showed a very significant inhibition of both the incidence and multiplicity of the liver tumors. These findings along with our initial reports suggest that betanin which is a regularly consumed natural product colorant is an effective cancer chemopreventive agent in mice. The most interesting observation is that the cancer chemopreventive effect was exhibited at a very low dose used in the study and thus indicating that beetroot warrants more attention for possible human applications in the control of malignancy. PMID- 12543063 TI - Oral erdosteine administration attenuates cisplatin-induced renal tubular damage in rats. AB - The effect of oral erdosteine on tissue malondialdehyde (MDA) and nitric oxide (NO) levels, and catalase (CAT), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities are investigated in the cisplatin model of acute renal failure in rats. A single dose of cisplatin caused kidney damage manifested by kidney histology as well as increases in plasma creatinine and blood urea nitrogen (BUN) levels. Treatment with free radical scavenger erdosteine attenuated increases in plasma creatinine and BUN, and tissue MDA and NO levels, and provided a histologically-proven protection against cisplatin-induced acute renal failure. Erdosteine also reduced depletion in the tissue CAT, GSH-Px, and SOD activities. These results show that erdosteine may be a promising drug for protection against cisplatin-induced nephrotoxicity. However, further studies with different doses of erdosteine are warranted for clarifying the issue. PMID- 12543064 TI - Different response of intracerebroventricular cadmium administration on blood pressure in normal and low urinary kallikrein rats. AB - Cadmium intracerebroventricular (i.c.v.) administration, at definite concentrations, induces a dose-dependent increase in the systemic blood pressure. Kallikreins are suggested to be important regulators of cardiovascular function. We evaluated the effects of 10 microg i.c.v. cadmium on mean blood pressure (MBP) and several urinary parameters such as 24h urine volume, sodium and potassium excretion and osmolality in a rat strain inbred for low urinary kallikrein and in normal-kallikrein Wistar rats. Low-kallikrein rats (LKR) showed an increase in MBP that, after an initial peak (27% from baseline), persisted higher than basal levels (10%) over 24h. In normal-kallikrein rats (NKR) a different reaction of blood pressure to cadmium was observed, causing a temporary increase (26% from baseline) of the systemic blood pressure, that returned to normal values within 2h. In addition, LKR showed a considerable reduction in the urinary volume (UV; 43.0+/-20 ml/24h versus 13.2+/-6 ml/24h, P<0.006), with an increase in the urinary osmolality (U(Osm); 500+/-210 mOsm/l versus 1391+/-245 mOsm/l, P<0.0002). Sodium (U(Na); 1761+/-432 microEq/24h versus 1156+/-522 microEq/24h, P<0.03) and potassium excretion (U(K); 2186+/-482 microEq/24h versus 936+/-299 microEq/24h, P<0.0006) were both significantly reduced. No changes in UV, U(Osm) and U(K) were observed in normal urinary kallikrein rats with the exception of U(Na) excretion that was significantly increased (667+/-274 microEq/24h versus 1725+/-300 microEq/24h, P<0.03). These results suggest that a genetically determined defect in urinary kallikrein excretion leads to a different hypertensive response to i.c.v. cadmium and to a different renal excretion of electrolytes. Perhaps the differences of blood pressure response could be due, at least in part, to a different sensitivity of LKR to cadmium: this implies a complex and articulate hypertensive effect of cadmium involving more systems than those supposed so far. PMID- 12543065 TI - Use of the proportionality equations for analyses of dose-response curves. AB - A proportionality-oriented theory was applied to analyze dose-response curves commonly generated in pharmacology. The principle of the proportionality theory is to express changes of two associated variables in reference to their asymptotes. Thus, a linear relationship between the two associated variables can be obtained if proper dimensions and scales are used. Based on this proportionality approach, we have developed equations which are used to analyze dose-response curves (1). generated by simulation data based on the Michaelis Menten equation and the Hill equation and (2). obtained from the contractile effect of acetylcholine (ACh) in isolated guinea pig ileum and the contractile effect of neurokinin NK(1) agonists (NK(1)) in guinea pig trachea muscle strips. Graphic methods are provided for plotting the graphs and for simultaneous determination of asymptote, slope parameter, and position constant. The slope parameter and position constant relate the concentration of an agonist to its response. Apparent equilibrium dissociation constant (K(A)), which is the product of position constant and asymptote in this approach, can be determined directly from the analysis of agonist dose-response curves. It is demonstrated that the proportionality theory and equations are useful for analyzing dose-response curves and for interpreting drug-receptor interactions. PMID- 12543066 TI - In vivo imaging. PMID- 12543067 TI - Quantitative motion analysis and visualization of cellular structures. AB - The availability of cellular markers tagged with the green fluorescent protein (GFP) has recently allowed a large number of cell biological studies to be carried out in live cells, thereby addressing the dynamic organization of cellular structures. Typically, microscopes capable of video recording are used to generate time-resolved data sets. Dynamic imaging data are complex and often difficult to interpret by pure visual inspection. Therefore, specialized image processing methods for object detection, motion estimation, visualization, and quantitation are required. In this review, we discuss concepts for automated analysis of multidimensional image data from live cell microscopy and their application to the dynamics of cell nuclear subcompartments. PMID- 12543068 TI - Using FRAP and mathematical modeling to determine the in vivo kinetics of nuclear proteins. AB - Fluorescence recovery after photobleaching (FRAP) has become a popular technique to investigate the behavior of proteins in living cells. Although the technique is relatively old, its application to studying endogenous intracellular proteins in living cells is relatively recent and is a consequence of the newly developed fluorescent protein-based living cell protein tags. This is particularly true for nuclear proteins, in which endogenous protein mobility has only recently been studied. Here we examine the experimental design and analysis of FRAP experiments. Mathematical modeling of FRAP data enables the experimentalist to extract information such as the association and dissociation constants, distribution of a protein between mobile and immobilized pools, and the effective diffusion coefficient of the molecule under study. As experimentalists begin to dissect the relative influence of protein domains within individual proteins, this approach will allow a quantitative assessment of the relative influences of different molecular interactions on the steady-state distribution and protein function in vivo. PMID- 12543069 TI - A high-speed multispectral spinning-disk confocal microscope system for fluorescent speckle microscopy of living cells. AB - Fluorescent speckle microscopy (FSM) uses a small fraction of fluorescently labeled subunits to give macromolecular assemblies such as the cytoskeleton fluorescence image properties that allow quantitative analysis of movement and subunit turnover. We describe a multispectral microscope system to analyze the dynamics of multiple cellular structures labeled with spectrally distinct fluorophores relative to one another over time in living cells. This required a high-resolution, highly sensitive, low-noise, and stable imaging system to visualize the small number of fluorophores making up each fluorescent speckle, a means by which to switch between excitation wavelengths rapidly, and a computer based system to integrate image acquisition and illumination functions and to allow a convenient interface for viewing multispectral time-lapse data. To reduce out-of-focus fluorescence that degrades speckle contrast, we incorporated the optical sectioning capabilities of a dual-spinning-disk confocal scanner. The real-time, full-field scanning allows the use of a low-noise, fast, high-dynamic range, and quantum-efficient cooled charge-coupled device (CCD) as a detector as opposed to the more noisy photomultiplier tubes used in laser-scanning confocal systems. For illumination, our system uses a 2.5-W Kr/Ar laser with 100-300mW of power at several convenient wavelengths for excitation of few fluorophores in dim FSM specimens and a four-channel polychromatic acousto-optical modulator fiberoptically coupled to the confocal to allow switching between illumination wavelengths and intensity control in a few microseconds. We present recent applications of this system for imaging the cytoskeleton in migrating tissue cells and neurons. PMID- 12543070 TI - Visualizing chromatin and chromosomes in living cells. AB - The dynamic organization of eukaryotic genomes in cell nuclei recently came into the focus of research interest. The kinetics of genome dynamics can be addressed only by approaches involving live cell microscopy. Different methods are available to visualize chromatin, specific chromatin fractions, or individual chromosome territories within nuclei of living mammalian cells. Appropriate labeling procedures as well as cell chamber systems and important controls for live cell microscopy are described. PMID- 12543071 TI - Visualizing RNA molecules inside the nucleus of living cells. AB - Fluorescence in situ hybridization is a widely used technique in cell biology providing insight into the spatial organization of specific RNA transcripts in the cell nucleus. However, to further investigate the dynamics of the transcription process and the transport rates of RNAs through the nucleus, RNAs need to be visualized and tracked in the living cell. In past years, various methods have been developed with the aim of tagging specific RNAs with a fluorescent moiety without interfering with cell vitality. These methods include the delivery of probes into a living cell, the in vivo hybridization of fluorescent oligonucleotide probes to endogenous RNAs, and the microscopic imaging of the tagged RNAs in living cells. In this article, we review a number of methods for tagging and visualizing endogenous RNAs in living cells. In addition, a protocol is described that allows detection of various RNA types using fluorochrome-labeled 2(')-O-methyl oligoribonucleotide (2(')-OMe RNA) probes. Compared with conventional oligodeoxynucleotide probes, 2(')-OMe RNA probes are not degraded by nucleases, form stable hybrids with structured RNAs, and do not interfere with cell vitality. PMID- 12543072 TI - Characterization of one- and two-photon excitation fluorescence resonance energy transfer microscopy. AB - Advances in molecular biology provide various methods to define the structure and function of the individual proteins that form the component parts of subcellular structures. The ability to see the dynamic behavior of a specific protein inside the living cell became possible through the application of advanced fluorescence resonance energy transfer (FRET) microscope techniques. The fluorophore molecule used for FRET imaging has a characteristic absorption and emission spectrum that should be considered for characterizing the FRET signal. In this article we describe the system development for the image acquisition for one- and two-photon excitation FRET microscopy. We also describe the precision FRET (PFRET) data analysis algorithm that we developed to remove spectral bleed-through and variation in the fluorophore expression level (or concentration) for the donor and acceptor molecules. The acquired images have been processed using a PFRET algorithm to calculate the energy transfer efficiency and the distance between donor and acceptor molecules. We implemented the software correction to study the organization of the apical endosome in epithelial polarized MDCK cells and dimerization of the CAATT/enhancer binding protein alpha (C/EBPalpha). For these proteins, the results revealed that the extent of correction affects the conventionally calculated energy transfer efficiency (E) and the distance (r) between donor and acceptor molecules by 38 and 9%, respectively. PMID- 12543073 TI - A dynamic view of cellular processes by in vivo fluorescence auto- and cross correlation spectroscopy. AB - Fluorescence correlation spectroscopy (FCS) is becoming increasingly popular as a technique that aims at complementing live cell images with biophysical information. This article provides both a short overview over recent intracellular FCS applications and a practical guide for investigators, who are seeking to integrate FCS into live cell imaging to obtain information on particle mobility, local concentrations, and molecular interactions. A brief introduction to the principles of FCS is provided, particularly emphasizing practical aspects such as the choice of appropriate dyes and positioning of the measurement volume in the sample. Possibilities and limitations in extracting parameters from autocorrelation curves are discussed, and attention is drawn to potential artifacts, such as photobleaching and probe aggregation. The principle of dual color cross-correlation is reviewed along with considerations for proper setup and adjustment. Practical implications of nonideal conditions including incomplete focus overlap and spectral cross-talk are considered. Recent examples of both auto- and cross-correlation applications demonstrate the potential of FCS for cell biology. PMID- 12543074 TI - The application of atomic force microscopy to the study of living vertebrate cells in culture. AB - Atomic force microscopy (AFM), a relatively new variant of scanning probe microscopy developed for the material sciences, is becoming an increasingly important tool in other disciplines. In this review I describe in nontechnical terms some of the basic aspects of using AFM to study living vertebrate cells. Although AFM has some unusual attributes such as an ability to be used with living cells, AFM also has attributes that make its use in cell biology a real challenge. This review was written to encourage researchers in the biological and biomedical sciences to consider AFM as a potential (and potent) tool for their cell biological research. PMID- 12543075 TI - Second harmonic generation imaging of endogenous structural proteins. AB - We show that structural protein arrays consisting largely of collagen, myosin, and tubulin, and their associated proteins can be imaged in three dimensions with high contrast and resolution by laser-scanning second harmonic generation (SHG) microscopy. SHG is a nonlinear optical scheme and this form of microscopy shares several common advantages with multiphoton excited fluorescence, namely, intrinsic three-dimensionality and reduced out-of-plane photobleaching and phototoxicity. SHG does not arise from absorption and in-plane photodamage considerations are therefore also greatly reduced. In particular, structural protein arrays that are highly ordered and birefringent produce large SHG signals without the need for any exogenous labels. We demonstrate that thick tissues including muscle and bone can be imaged and sectioned through several hundred micrometers of depth. Combining SHG with two-photon excited green fluorescent protein (GFP) imaging allows inference of the molecular origin of the SHG contrast in Caenorhabditis elegans sarcomeres. Symmetry and organization of microtubule structures in dividing C. elegans embryos are similarly studied by comparing the endogenous tubulin contrast with that of GFP::tubulin fluorescence. It is found that SHG provides molecular level data on radial and lateral symmetries that GFP constructs cannot. The physical basis of SHG is discussed and compared with that of two-photon excitation as well as that of polarization microscopy. Due to the intrinsic sectioning, lack of photobleaching, and availability of molecular level data, SHG is a powerful tool for in vivo imaging. PMID- 12543076 TI - Visualizing protein-protein interactions in living animals. AB - A variety of techniques have been developed to analyze protein-protein interactions in vitro and in cultured cells. However, these methods do not determine how protein interactions affect and are regulated by physiologic and pathophysiologic conditions in living animals. This article describes methodology for detecting and quantifying protein interactions in living mice, using an inducible two-hybrid system developed for positron emission tomography (PET) imaging. We discuss the methods to establish stably transfected cells with components of the imaging system, create tumor xenografts, synthesize PET radiopharmaceuticals used to visualize the imaging reporter, perform microPET imaging, and analyze data from imaging studies. Development and application of technologies for molecular imaging of protein-protein interactions in vivo should enable researchers to investigate intrinsic binding specificities of proteins during normal development and disease progression as well as aid drug development through direct interrogation of molecular targets within intact animals. PMID- 12543077 TI - SHP2 regulates IL-2 induced MAPK activation, but not Stat3 or Stat5 tyrosine phosphorylation, in cutaneous T cell lymphoma cells. AB - The phosphotyrosine phosphatase SHP2 has been suggested to regulate activation of MAPK, Stat3, and Stat5 in several experimental models. In this study we investigated the role of SHP2 in IL-2 induced activation of MAPK and the Stat proteins using the human CTCL cell line MyLa2059 derived from a cutaneous T cell lymphoma (CTCL). For this purpose, MyLa2059 cells were stably transfected with wild-type SHP2 or inactive SHP2. The cells transfected with inactive SHP2 showed reduced MAPK activation upon IL-2 stimulation, suggesting that SHP2 upregulates IL-2 induced MAPK activation in T cells. However, the constitutive tyrosine phosphorylation of Stat3 as well as IL-2 induced Stat5 tyrosine phosphorylation and DNA binding were unaffected by the stably transfected wild-type SHP2 as well as the inactive SHP2. In conclusion, we show for the first time that SHP2 positively regulates IL-2 induced MAPK activation in malignant T cells. Furthermore, the results indicate that SHP2 may not be involved in the activation of Stat3 or Stat5 in CTCL cells. PMID- 12543079 TI - Effect of azacytidine in the release of leukemia inhibitory factor, oncostatin m, interleukin (IL)-6, and IL-11 by mononuclear cells of patients with refractory anemia. AB - 5-azacytidine (AZA) yields hematologic improvement in patients with myelodysplastic syndromes (MDS). Ineffective hemopoiesis in MDS produce the paradox of high intramedullary cellularity with peripheral cytopenias. Leukemia inhibitory factor (LIF), oncostatin M (OSM), interleukin (IL)-6, and IL-11 regulate hemopoiesis and LIF, OSM, and IL-6 also inhibit the proliferation of myeloid leukemic cell lines through the signal-transducing subunit gp130. These IL-6-type cytokines were measured by enzyme-linked immunosorbent assay in cell culture supernatants (SN) obtained from peripheral blood mononuclear cells (MNC) and monocyte-depleted MNC of patients with refractory anemia (RA; n=12) and healthy individuals (n=10). AZA down-regulated OSM, IL-6, and IL-11 release by MNC of patients but not by MNC from healthy individuals. Patient's SN had significantly lower concentrations of LIF, OSM, and IL-11 than SN of normal subjects. When monocyte-depleted MNC of patients were stimulated with phytohemagglutinin a significant increment in OSM levels was observed. In contrast, monocyte depletion in healthy subjects did not cause any significant change in OSM values. We conclude that: (a) AZA inhibits the release of OSM, IL 6, and IL-11 exclusively in RA-diseased MNC, (b) Patient's MNC release subnormal amounts of LIF, OSM, and IL-11, and (c) RA-derived monocytes probably down regulate OSM release by phytohemagglutinin-activated MNC. PMID- 12543078 TI - Signaling pathways for TNF production induced by human aminoacyl-tRNA synthetase associating factor, p43. AB - The p43 protein is associated with human macromolecular aminoacyl tRNA synthetase complex and secreted to up-regulate diverse proinflammatory genes including TNF. Here we focused on the p43-induced TNF production and determined its responsible signal pathway. The p43-induced TNF production was mediated by the activation of MAPK family members, ERK and p38 MAPK, and by IkappaB degradation leading to the activation of NFkappaB. We also studied the upstream molecules for ERK and p38 MAPK by using a variety of inhibitors. The inhibitors for protein kinase C (PKC) and phospholipase C (PLC) prevented the p43-induced TNF production. Interestingly, all of the effective drugs inhibited the ERK activity, while the drugs had no effects on p38 MAPK activity and IkappaB degradation. Together, the p43-induced TNF production was controlled by NFkB, p38 MAPK, and ERK that is dependent on the activities of PLC and PKC. PMID- 12543080 TI - Modulation by colony stimulating factors of human epithelial colon cancer cell apoptosis. AB - Colony stimulating factors (CSF) promote leukocyte survival by reducing apoptotic cell death. However, their effects on non-leukocyte cell types are unclear. Reduced apoptosis in colon epithelial cells is thought to contribute to the initiation of cancer. Here, we report diminished spontaneous apoptosis of human colon epithelial HT-29 cells in the presence of macrophage-CSF or granulocyte macrophage-CSF. Moreover, reduced apoptosis induced by sulindac sulfide was also observed with macrophage-CSF. Granulocyte-CSF failed to modify spontaneous or sulindac sulfide induced apoptosis. It seems, therefore, that the action of CSFs on apoptosis is not confined to haematopoietic cells but may be extended to stromal cells. PMID- 12543081 TI - Morphological analyses of interleukin-8 effects on rat ovarian follicles at ovulation and luteinization in vivo. AB - The aim of the present study was to elucidate functions of the interleukin (IL)-8 at ovulation and luteinization in vivo. To compare the morphological differences between human chorionic gonadotropin (hCG) and IL-8 stimulation, scanning electron microscopy was employed to study rat ovarian vascular corrosion casts. Follicular growth and increased capillary vessel densities around the follicles were seen in vascular corrosion casts after IL-8 injection, similar to the result of hCG administration. This result indicated that exogenous IL-8 could play a role in the neovascularization during follicular development as an angiogenetic factor. Many fenestrations were observed in the vascular endothelium by hCG administration. In contrast, no fenestrations were observed with IL-8 injection, indicating that IL-8 may not be sufficient to increase the vascular permeability directly. Although germinal vesicle breakdown (GVBD) occurred at rates of 82% after the hCG injection, only 20% GVBD was observed after the IL-8 injection. The present study indicated that IL-8 might have important effects on rat follicles at ovulation and luteinization via vascularization in a similar manner to hCG. However, IL-8 was not effective on vascular permeability and oocyte maturation, which were different from hCG. Thus, we can conclude that IL-8 can participate in follicular development in part and may play important roles in ovulation and luteinization as one of some mediators induced by endogenous luteinizing hormone. PMID- 12543082 TI - A longitudinal study of leptin and appetite, resting energy expenditure and body fat mass in weight-stable cancer patients. AB - Leptin and its relationship with energy metabolism in male weight-stable patients with colorectal liver metastases (n=14) was assessed at baseline and after 6 weeks. At baseline, median leptin concentration was 5.9 microg/l and the median percentage fat mass was 32.1%. Circulating leptin concentrations were correlated with measured percentage fat mass at baseline (r(s)=0.519, P=0.040) and with the changes after 6 weeks (r(s)=0.611, P=0.027) but not with insulin, cortisol, C reactive protein, appetite or resting energy expenditure. Therefore, it would appear that leptin concentrations reflect changes in fat mass in male weight stable patients with cancer and their role in the regulation of energy metabolism appears more complex than previously proposed. PMID- 12543083 TI - Abnormal production of interleukin (IL)-11 and IL-8 in polycythaemia vera. AB - We studied the production of interleukin (IL)-11 and IL-8, two cytokines known to affect erythropoiesis, in polycythemia vera (PV). In vivo, IL-11 was detected more frequently in serum and bone marrow (BM) plasma of PV patients than in controls (healthy donors and patients with idiopathic erythrocytosis (IE)). In addition, serum IL-11 levels of PV patients were higher than those of controls. IL-8 was elevated in serum of both PV and IE patients (respective median levels: 38.6 and 242pg/ml, vs 4.4pg/ml for healthy donors). BM plasma IL-8 levels of PV patients (508pg/ml) were significantly higher than those of IE patients (120pg/ml). In vitro, bone marrow (BM) stromal cells (BMSC) of PV patients produced significantly more IL-11 (x6.4) and IL-8 (x8.3) than BMSC of healthy donors or IE patients. In conclusion, both IL-11 and IL-8 are overproduced in PV, apparently by BMSC; IL-8 is also overproduced in IE, by cells other than BMSC. PMID- 12543084 TI - Prognostic role of serum vascular endothelial growth factor, basic fibroblast growth factor and nitric oxide in patients with colorectal carcinoma. AB - Recently, angiogenesis has gained an increasing interest as a prognostic factor in a variety of solid tumours. In this study we aimed to assess the prognostic role of serum vascular endothelial growth factor (VEGF), basic fibroblast growth factor (b-FGF) and nitric oxide (NO) levels in patients with colorectal carcinoma (CRC).A total of 52 consecutive colorectal cancer patients with stage I to IV disease was included. In addition to routine laboratory and staging procedures, serum VEGF, b-FGF levels, and nitrate levels as a surrogate marker for in-vivo NO production were assayed. Serum VEGF concentrations, adjusted to the platelet count were found to be a significant factor for overall survival in univariate analysis (P=0.033). A new angiogenic index (AI), derived from serum VEGF and nitrate concentrations, was established. AI is the only independent prognostic factor of survival in all patients (P=0.008, Cox regression analysis). Likewise, AI is also significant prognostic factor for disease-free survival (DFS) in patients with operable CRC (P=0.032, Cox regression analysis). In conclusion, serum VEGF and NO levels have prognostic role in patients with CRC and the new angiogenesis index using the serum levels of the factors seem to be useful. PMID- 12543085 TI - The endoplasmic reticulum is a focal point for co-ordination of cellular activity. PMID- 12543086 TI - The endoplasmic reticulum: a multifunctional signaling organelle. AB - The endoplasmic reticulum (ER) is a multifunctional signaling organelle that controls a wide range of cellular processes such as the entry and release of Ca(2+), sterol biosynthesis, apoptosis and the release of arachidonic acid (AA). One of its primary functions is as a source of the Ca(2+) signals that are released through either inositol 1,4,5-trisphosphate (InsP(3)) or ryanodine receptors (RYRs). Since these receptors are Ca(2+)-sensitive, the ER functions as an excitable system capable of spreading signals throughout the cell through a process of Ca(2+)-induced Ca(2+) release (CICR). This regenerative capacity is particularly important in the control of muscle cells and neurons. Its role as an internal reservoir of Ca(2+) must be accommodated with its other major role in protein synthesis where a constant luminal level of Ca(2+) is essential for protein folding. The ER has a number of stress signaling pathways that activate various transcriptional cascades that regulate the luminal content of the Ca(2+) dependent chaperones responsible for the folding and packaging of secretory proteins.Another emerging function of the ER is to regulate apoptosis by operating in tandem with mitochondria. Anti-apoptotic regulators of apoptosis such as Bcl-2 may act by reducing the ebb and flow of Ca(2+) through the ER/mitochondrial couple. Conversely, the presenilins that appear to increase the Ca(2+) content of the ER lumen make cells more susceptible to apoptosis. PMID- 12543087 TI - Measuring [Ca2+] in the endoplasmic reticulum with aequorin. AB - The photoprotein aequorin was the first probe used to measure specifically the [Ca(2+)] inside the lumen of the endoplasmic reticulum ([Ca(2+)](ER)) of intact cells and it provides values for the steady-state [Ca(2+)](ER), around 500 microM, that closely match those obtained now by other procedures. Aequorin-based methods to measure [Ca(2+)](ER) offer several advantages: (i) targeting of the probe is extremely precise; (ii) the use of low Ca(2+)-affinity aequorin allows covering a large dynamic range of [Ca(2+)], from 10(-5) to 10(-3)M; (iii) aequorin is nearly insensitive to changes in Mg(2+) or pH, has a high signal-to noise ratio and calibration of the results in [Ca(2+)] is made straightforward using a simple algorithm; and (iv) the equipment required for luminescence measurements in cell populations is simple and low-cost. On the negative side, this technique has also some disadvantages: (i) the relatively low amount of emitted light makes difficult performing single-cell imaging studies; (ii) reconstitution of aequorin with coelenterazine requires previous complete depletion of Ca(2+) of the ER for 1-2h, a maneuver that may result in deleterious effects in some cells; (iii) because of the high rate of aequorin consumption at steady-state [Ca(2+)](ER), only relatively brief experiments can be performed; and (iv) expression of ER-targeted aequorin requires previous transfection or infection to introduce the appropriate DNA construct, or alternatively the use of stable cell clones. Choosing aequorin or other techniques to measure [Ca(2+)](ER) will depend of the correct balance between these properties in a particular problem. PMID- 12543088 TI - The distribution of the endoplasmic reticulum in living pancreatic acinar cells. AB - Studies on pancreatic acinar cells provided the original evidence for the Ca(2+) releasing action of inositol 1,4,5-trisphosphate (IP(3)). Ironically, this system has presented problems for the general theory that IP(3) acts primarily on the endoplasmic reticulum (ER), because the IP(3)-elicited Ca(2+) release occurs in the apical pole, which is dominated by zymogen granules (ZGs) and apparently contains very little ER. Using confocal and two-photon microscopy and a number of different ER-specific fluorescent probes, we have now investigated in detail the distribution of the ER in living pancreatic acinar cells. It turns out that although the bulk of the ER, as expected, is clearly located in the baso-lateral part of the cell, there is significant invasion of ER into the granular pole and each ZG is in fact surrounded by strands of ER. This structural evidence from living cells, in conjunction with recent functional studies demonstrating the high Ca(2+) mobility in the ER lumen, provides the framework for a coherent and internally consistent theory for cytosolic Ca(2+) signal generation in the apical secretory pole, in which the primary Ca(2+) release occurs from ER extensions in the granular pole supplied with Ca(2+) from the main store at the base of the cell by the tunnel function of the ER. PMID- 12543089 TI - Ca2+ signaling and calcium binding chaperones of the endoplasmic reticulum. AB - The endoplasmic reticulum is a centrally located organelle which affects virtually every cellular function. Its unique luminal environment consists of Ca(2+) binding chaperones, which are involved in protein folding, post translational modification, Ca(2+) storage and release, and lipid synthesis and metabolism. The environment within the lumen of the endoplasmic reticulum has profound effects on endoplasmic reticulum function and signaling, including apoptosis, stress responses, organogenesis, and transcriptional activity. Calreticulin, a major Ca(2+) binding (storage) chaperone in the endoplasmic reticulum, is a key component of the calreticulin/calnexin cycle which is responsible for the folding of newly synthesized proteins and glycoproteins and for quality control pathways in the endoplasmic reticulum. The function of calreticulin, calnexin and other endoplasmic reticulum proteins is affected by continuous fluctuations in the concentration of Ca(2+) in the endoplasmic reticulum. Thus, changes in Ca(2+) concentration may play a signaling role in the lumen of the endoplasmic reticulum as well as in the cytosol. Recent studies on calreticulin-deficient and transgenic mice have revealed that calreticulin and the endoplasmic reticulum may be upstream regulators in the Ca(2+)-dependent pathways that control cellular differentiation and/or organ development. PMID- 12543090 TI - Molecular physiology of the SERCA and SPCA pumps. AB - Intracellular Ca(2+)-transport ATPases exert a pivotal role in the endoplasmic reticulum and in the compartments of the cellular secretory pathway by maintaining a sufficiently high lumenal Ca(2+) (and Mn(2+)) concentration in these compartments required for an impressive number of vastly different cell functions. At the same time this lumenal Ca(2+) represents a store of releasable activator Ca(2+) controlling an equally impressive number of cytosolic functions. This review mainly focuses on the different Ca(2+)-transport ATPases found in the intracellular compartments of mainly animal non-muscle cells: the sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA) pumps. Although it is not our intention to treat the ATPases of the specialized sarcoplasmic reticulum in depth, we can hardly ignore the SERCA1 pump of fast-twitch skeletal muscle since its structure and function is by far the best understood and it can serve as a guide to understand the other members of the family. In a second part of this review we describe the relatively novel family of secretory pathway Ca(2+)/Mn(2+) ATPases (SPCA), which in eukaryotic cells are primarily found in the Golgi compartment. PMID- 12543091 TI - Molecular genetics of ryanodine receptors Ca2+-release channels. AB - The family of ryanodine receptor (RyR) genes encodes three highly related Ca(2+) release channels: RyR1, RyR2 and RyR3. RyRs are known as the Ca(2+)-release channels that participate to the mechanism of excitation-contraction coupling in striated muscles, but they are also expressed in many other cell types. Actually, in several cells two or three RyR isoforms can be co-expressed and interactive feedbacks among them may be important for generation of intracellular Ca(2+) signals and regulation of specific cellular functions. Important developments have been obtained in understanding the biochemical complexity underlying the process of Ca(2+) release through RyRs. The 3-D structure of these large molecules has been obtained and some regulatory regions have been mapped within these 3-D reconstructions. Recent studies have clarified the role of protein kinases and phosphatases that, by physically interacting with RyRs, appear to play a role in the regulation of these Ca(2+)-release channels. These and other recent advancements in understanding RyR biology will be the object of this review. PMID- 12543092 TI - IP3 receptors and their regulation by calmodulin and cytosolic Ca2+. AB - Inositol 1,4,5-trisphosphate (IP(3)) receptors are tetrameric intracellular Ca(2+) channels, the opening of which is regulated by both IP(3) and Ca(2+). We suggest that all IP(3) receptors are biphasically regulated by cytosolic Ca(2+), which binds to two distinct sites. IP(3) promotes channel opening by controlling whether Ca(2+) binds to the stimulatory or inhibitory sites. The stimulatory site is probably an integral part of the receptor lying just upstream of the pore region. Inhibition of IP(3) receptors by Ca(2+) probably requires an accessory protein, which has not yet been unequivocally identified, but calmodulin is a prime candidate. We speculate that one lobe of calmodulin tethers it to the IP(3) receptor, while the other lobe can bind Ca(2+) and then interact with a second site on the receptor to cause inhibition. PMID- 12543093 TI - Sphingosine 1-phosphate: a Ca2+ release mediator in the balance. AB - Sphingosine 1-phosphate (S1P) is a lipid signalling molecule with Ca(2+) mobilising properties. Importantly for a role as a Ca(2+) release messenger, intracellular levels of S1P can be regulated by a variety of extracellular stimuli, via the enzyme sphingosine kinase. However, neither the mechanism underlying S1P generation, nor its actions at the endoplasmic reticulum are clear. Thus, the role of S1P as an intracellular mediator of Ca(2+) release remains in the balance. PMID- 12543094 TI - Interactions between calcium release pathways: multiple messengers and multiple stores. AB - The discovery of cyclic adenosine diphosphate ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP) as Ca(2+) releasing messengers has provided additional insight into how complex Ca(2+) signalling patterns are generated. There is mounting evidence that these molecules along with the more established messenger, myo-inositol 1,4,5-trisphosphate (IP(3)), have a widespread messenger role in shaping Ca(2+) signals in many cell types. These molecules have distinct structures and act on specific Ca(2+) release mechanisms. Emerging principles are that cADPR enhances the Ca(2+) sensitivity of ryanodine receptors (RYRs) to produce prolonged Ca(2+) signals through Ca(2+)-induced Ca(2+) release (CICR), while NAADP acts on a novel Ca(2+) release mechanism to produce a local trigger Ca(2+) signal which can be amplified by CICR by recruiting other Ca(2+) release mechanisms. Whilst IP(3) and cADPR mobilise Ca(2+) from the endoplasmic reticulum (ER), recent evidence from the sea urchin egg suggests that the major NAADP-sensitive Ca(2+) stores are reserve granules, acidic lysosomal-related organelles. In this review we summarise the role of multiple Ca(2+) mobilising messengers, Ca(2+) release channels and Ca(2+) stores, and the interplay between them, in the generation of specific Ca(2+) signals. Focusing upon cADPR and NAADP, we discuss how cellular stimuli may draw upon different combinations of these messengers to produce distinct Ca(2+) signalling signatures. PMID- 12543095 TI - Calcium leak from intracellular stores--the enigma of calcium signalling. AB - Wherever you travel through the cytoplasm of the cells you will find organelles with internal [Ca(2+)] levels higher than in the surrounding cytosol. This is particularly true of the endoplasmic reticulum (ER) (or sarcoplasmic reticulum (SR) in muscle cells); such organelles serve as the main sources of releasable Ca(2+) for cytosolic cellular signalling. Calcium pumps of the SERCA family (sarcoplasmic and endoplasmic reticulum calcium ATP-ases) import calcium into the organelle lumen. The other mechanism that is responsible for the steady state calcium level within the lumen of ER or SR is a calcium leak that balances the influx created by the pumps. The leak remains the most enigmatic of the processes involved in calcium regulation. The molecular nature of the leak mechanism is not known. The basal leak is a relatively slow process, which is difficult to investigate and which is easily outmatched (both in the amplitude of calcium responses and in attractiveness to experimenters) by substantially faster second messenger-induced release. Nevertheless, information on the properties of the calcium leak, although thinly scattered through the pages of PubMed, has been slowly accumulating. In this review we will discuss the properties of the calcium leak and speculate about possible mechanisms, which could mediate this process. PMID- 12543096 TI - Old players in a new role: mitochondria-associated membranes, VDAC, and ryanodine receptors as contributors to calcium signal propagation from endoplasmic reticulum to the mitochondria. AB - In many cell types, IP(3) and ryanodine receptor (IP(3)R/RyR)-mediated Ca(2+) mobilization from the sarcoendoplasmic reticulum (ER/SR) results in an elevation of mitochondrial matrix [Ca(2+)]. Although delivery of the released Ca(2+) to the mitochondria has been established as a fundamental signaling process, the molecular mechanism underlying mitochondrial Ca(2+) uptake remains a challenge for future studies. The Ca(2+) uptake can be divided into the following three steps: (1) Ca(2+) movement from the IP(3)R/RyR to the outer mitochondrial membrane (OMM); (2) Ca(2+) transport through the OMM; and (3) Ca(2+) transport through the inner mitochondrial membrane (IMM). Evidence has been presented that Ca(2+) delivery to the OMM is facilitated by a local coupling between closely apposed regions of the ER/SR and mitochondria. Recent studies of the dynamic changes in mitochondrial morphology and visualization of the subcellular pattern of the calcium signal provide important clues to the organization of the ER/SR mitochondrial interface. Interestingly, key steps of phospholipid synthesis and transfer to the mitochondria have also been confined to subdomains of the ER tightly associated with the mitochondria, referred as mitochondria-associated membranes (MAMs). Through the OMM, the voltage-dependent anion channels (VDAC, porin) have been thought to permit free passage of ions and other small molecules. However, recent studies suggest that the VDAC may represent a regulated step in Ca(2+) transport from IP(3)R/RyR to the IMM. A novel proposal regarding the IMM Ca(2+) uptake site is a mitochondrial RyR that would mediate rapid Ca(2+) uptake by mitochondria in excitable cells. An overview of the progress in these directions is described in the present paper. PMID- 12543097 TI - Permeation through store-operated CRAC channels in divalent-free solution: potential problems and implications for putative CRAC channel genes. AB - CRAC channels are key calcium conduits in both physiological and pathological states. Understanding how these channels are controlled is important as this will not only provide insight into a novel signal transduction pathway coupling intracellular stores to the channels in the plasma membrane, but might also be of clinical relevance. Determining the molecular identity of the CRAC channels will certainly be a major step forward. Like all Ca(2+)-selective channels, CRAC channels lose their selectivity in divalent-free external solution to support large monovalent Na(+) currents. This approach has provided new insight into channel permeation and selectivity, and identifies some interesting differences between CRAC channels and voltage-operated calcium channels (VOCCs). Studies in divalent-free solution are a double-edged sword, however. Electrophysiologists need to be wary because some of the conditions used to study I(CRAC) in divalent free external solution, notably omission of Mg(2+)/Mg-ATP from the recording pipette solution, activates an additional current permeating through Mg(2+) nucleotide-regulated metal ion current (MagNuM; TRPM7) channels. This channel underlies the large single-channel events that have been attributed to CRAC channels in the past and which have been used to as a tool to identify store operated channels in native cells and recombinant expression systems.Are we any closer to identifying the elusive CRAC channel gene(s)? TRPV6 seemed a very attractive candidate, but one of the main arguments supporting it was a single channel conductance in divalent-free solution similar to that for CRAC reported under conditions where MagNuM is active. We now know that the conductance of TRPV6 is approximately 200-fold larger than that of CRAC in native tissue. Moreover, it is unclear if TRPV6 is store-operated. Further work on TRPV6, particularly whether its single-channel conductance is still high under conditions where it apparently forms multimers with endogenous store-operated channels, and whether it is activated by a variety of store depletion protocols, will be helpful in finally resolving this issue. PMID- 12543098 TI - The endoplasmic reticulum and neuronal calcium signalling. AB - The endoplasmic reticulum (ER) is a multifunctional signalling organelle regulating a wide range of neuronal functional responses. The ER is intimately involved in intracellular Ca(2+) signalling, producing local or global cytosolic calcium fluctuations via Ca(2+)-induced Ca(2+) release (CICR) or inositol-1,4,5 trisphosphate-induced Ca(2+) release (IICR). The CICR and IICR are controlled by two subsets of Ca(2+) release channels residing in the ER membrane, the Ca(2+) gated Ca(2+) release channels, generally known as ryanodine receptors (RyRs) and InsP(3)-gated Ca(2+) release channels, referred to as InsP(3)-receptors (InsP(3)Rs). Both types of Ca(2+) release channels are expressed abundantly in nerve cells and their activation triggers cytoplasmic Ca(2+) signals important for synaptic transmission and plasticity. The RyRs and InsP(3)Rs show heterogeneous localisation in distinct cellular sub-compartments, conferring thus specificity in local Ca(2+) signals. At the same time, the ER Ca(2+) store emerges as a single interconnected pool fenced by the endomembrane. The continuity of the ER Ca(2+) store could play an important role in various aspects of neuronal signalling. For example, Ca(2+) ions may diffuse within the ER lumen with comparative ease, endowing this organelle with the capacity for "Ca(2+) tunnelling". Thus, continuous intra-ER Ca(2+) highways may be very important for the rapid replenishment of parts of the pool subjected to excessive stimulation (e.g. in small compartments within dendritic spines), the facilitated removal of localised Ca(2+) loads, and finally in conveying Ca(2+) signals from the site of entry towards the cell interior and nucleus. PMID- 12543099 TI - Calcium stores and synaptic plasticity. AB - Chemical transmission at central synapses is known to be highly plastic; the strength of synaptic connections can be modified bi-directionally as a result of activity at individual synapses. Long-term changes in synaptic efficacy, both increases and decreases, are thought to be involved in the development of the nervous system, and in ongoing changes in response to external cues such as during learning and addiction. Other, shorter lasting changes in synaptic transmission are also likely to be important in normal functioning of the CNS. Calcium mobilisation is an important step in multiple forms of plasticity and, although entry into neurones from the extracellular space is often the initial trigger for plasticity changes, release of calcium from intracellular stores also has an important part to play in a variety of forms of synaptic plasticity. PMID- 12543100 TI - Endoplasmic reticulum, Bcl-2 and Ca2+ handling in apoptosis. AB - In the complex signalling interplay that allows extracellular signals to be decoded into activation of apoptotic cell death, Ca(2+) plays a significant role. This is supported not only by evidence linking alterations in Ca(2+) homeostasis to the triggering of apoptotic (and in some cases necrotic) cell death, but also by recent data indicating that a key anti-apoptotic protein, Bcl-2, has a direct effect on ER Ca(2+) handling. We will briefly summarise the first aspect, and describe in more detail these new data, demonstrating that (i) Bcl-2 reduces the state of filling of the ER Ca(2+) store and (ii) this Ca(2+) signalling alteration renders the cells less sensitive to apoptotic stimuli. Overall, these results suggest that calcium homeostasis may represent a pharmacological target in the fundamental pathological process of apoptosis. PMID- 12543101 TI - Neuronal survival in the balance: are endoplasmic reticulum membrane proteins the fulcrum? PMID- 12543102 TI - Allogeneic hematopoietic stem cell transplantation--yesterday, today, and tomorrow. AB - During the past 50 years, intensive studies into the use of hematopoietic stem cell transplantation (HSCT) for therapy of cancer and nonmalignant hematologic diseases have changed this treatment modality from one that was thought to be plagued by insurmountable complications to one that is now standard therapy for some diseases. More-recent research has opened up the way to include elderly patients with a wide variety of hematologic malignant diseases. Continued research by transplant teams worldwide is likely to allow continued progress toward developing novel and improved treatment modalities and even wider application of the use of pluripotent hematopoietic stem cells in the treatment of human diseases. PMID- 12543103 TI - Combinatorial peptide library methods for immunobiology research. AB - The field of combinatorial peptide chemistry has emerged as a powerful tool in the study of many biological systems. This review focuses on combinatorial peptide library methodology, which includes biological library methods, spatially addressable parallel library methods, library methods requiring deconvolution, the "one-bead one-compound" library method, and affinity chromatography selection method. These peptide libraries have successfully been employed to study a vast array of cell surface receptors, as well as have been useful in identifying protein kinase substrates and inhibitors. In recent immunobiological applications, peptide libraries have proven monumental in the definition of MHC anchor residues, in lymphocyte epitope mapping, and in the development of peptide vaccines. Peptides identified from such libraries, when presented in a chemical microarray format, may prove useful in immunodiagnostics. Combinatorial peptide libraries offer a high-throughput approach to study limitless biological targets. Peptides discovered from such studies may be therapeutically and diagnostically useful agents. PMID- 12543104 TI - Human CD34(+) blood cells induce T-cell unresponsiveness to specific alloantigens only under costimulatory blockade. AB - OBJECTIVES: The immunogenic role of human CD34(+) cells in allogeneic hematopoietic stem cell transplantation is controversial. In this study we tested the role of CD40 and CTLA4 ligands on CD34(+) cell costimulation of HLA mismatched lymphocytes. MATERIALS AND METHODS: An anti-CD40L monoclonal antibody (hu5C8) and/or CTLA4-Ig molecule were used in primary mixed lymphocyte culture (MLC) with irradiated CD34(+) blood cells and allogeneic responders. Then, secondary MLC, cytotoxic activity, and effector cytokine expression and production were measured. RESULTS: Each reagent was able to reduce anti-CD34(+) cell alloreactivity, but only the combination of the anti-CD40L monoclonal antibody and CTLA4-Ig induced greater than 90% inhibition of T-cell response in primary MLC and prevented generation of cytotoxic T cells when priming with purified CD34(+) cells. Importantly, responder cells activated by allogeneic CD34(+) cells in the presence of anti-CD40L monoclonal antibody and CTLA4-Ig entered a state of antigen-specific unresponsiveness while responding to third party antigen, tetanus toxoid, or phytohemagglutinin, and showed suppression of interferon-gamma and increase of interleukin-10 expression and release. Interestingly, addition of interleukin-2 in secondary MLC did not reverse T-cell anergy. CONCLUSIONS: The results demonstrate that human CD34(+) blood progenitors stimulate T-cell responses potently and can induce T-cell unresponsiveness only when both B7:CD28 and CD40:CD40L pathways are blocked, with an increase of interleukin-10-producing cells. Therefore, our data allow design of in vivo studies aimed at achieving T-cell tolerance across HLA barriers by using purified CD34(+) cells and costimulatory blockade. PMID- 12543105 TI - Transient expression of PU.1 commits multipotent progenitors to a myeloid fate whereas continued expression favors macrophage over granulocyte differentiation. AB - OBJECTIVES: The Ets-family transcription factor PU.1 is expressed specifically in the hematopoietic system, in which it is absolutely required for the generation of B lymphocytes and macrophages. In contrast, overexpression of PU.1 blocks terminal differentiation of the erythroid lineage, in which it can act as an oncogene. In this study we used a multipotential progenitor cell line to examine the effects of PU.1 overexpression on myeloerythroid commitment within a single model system. MATERIALS AND METHODS: PU.1 cDNA was introduced transiently and stably into the multipotent, nonleukemic hemopoietic cell line FDCPmix. Transiently transfected cells were isolated by fluorescence-activated cell sorting within 18 hours of transfection. Stable transfectants were selected by antibiotic resistance over a number of weeks. The effects of short- and long-term overexpression of PU.1 on self-renewal, proliferation, and differentiation were investigated. RESULTS: A transient pulse of expression in multipotent progenitor cells eliminated the options of self-renewal and erythroid differentiation, resulting in commitment to the myeloid lineage. However, this transient pulse of expression did not affect the subsequent lineage choice of bipotent granulocyte/macrophage progenitors. In contrast, continuous expression of PU.1 resulted in a strong bias toward macrophage rather than granulocyte differentiation. CONCLUSIONS: These results demonstrate promyeloid effects of PU.1 at two distinct stages of hematopoiesis. PMID- 12543106 TI - Selective in vivo growth of lymphocyte function- associated antigen-1-positive murine myeloma cells. Involvement of function-associated antigen-1-mediated homotypic cell-cell adhesion. AB - OBJECTIVES: Lymphocyte function-associated antigen-1 (LFA-1) expression on multiple myeloma cells and its potential role in myeloma biology have been the subject of conflicting literature reports. In this study we used the 5T experimental mouse model to analyze the involvement of LFA-1 in myeloma cell bone marrow homing, survival, and growth. MATERIALS AND METHODS: The 5T33MM vitro (5T33MMvt) myeloma line was used. LFA-1 and intercellular adhesion molecule-1 (ICAM-1) expression were analyzed by flow cytometry. A small molecule antagonist of LFA-1/ICAM interactions, BIRT 377, was used to block LFA-1 in vitro. Transendothelial migration was assessed by measuring migration through Transwells coated with bone marrow endothelial cells. Immediate in vivo homing was analyzed by tracing 51Cr-labeled cells. Invert microscopic cell counting was used to analyze homotypic cell adhesion. Cell cycle analysis was used to analyze apoptosis. S+G(2)/M phase analysis and 3H-thymidine incorporation were used to assess proliferation. Cells were separated into LFA-1(+) and LFA-1(-) fraction by magnetic activated cell sorting. RESULTS: 5T33MMvt cells had a heterogeneous LFA 1 expression and all cells were positive for the LFA-1 ligand ICAM-1. LFA-1 inhibition with BIRT 377 did not affect transendothelial migration of the 5T33MMvt cells; however, it did result in cell cluster scattering, indicating LFA 1 involvement in homotypic cell-cell adhesion. No effect was observed on apoptosis, but the percentage of cells in S+G(2)/M phase was decreased by 39%. 3H thymidine incorporation confirmed this effect on 5T33MMvt cell proliferation (38% reduction). When 5T33MMvt cells were injected into animals, all myeloma cells isolated at the end stage of the disease were LFA-1(+) in contrast to the situation before injection. LFA-1(+) and LFA-1(-) MM cells had similar in vivo bone marrow homing capacities. Mice injected with LFA-1(+) 5T33MMvt cells developed myeloma (5/5) within 12 weeks after injection. In contrast, LFA-1(-) recipients did not develop the disease (0/5), even 1 year after tumor inoculation. CONCLUSIONS: Our data suggest that LFA-1-mediated homotypic cell cell adhesion is involved in myeloma cell proliferation and raises the possibility that this interaction may have a crucial role in in vivo myeloma cell growth. LFA-1 does not appear to play a role in the bone marrow homing of these cells. PMID- 12543107 TI - Using divisional history to measure hematopoietic stem cell self-renewal and differentiation. AB - OBJECTIVES: The purpose of this study was to investigate cell fates and long-term repopulating potential of a primitive hematopoietic stem cell (HSC) population (i.e., FR25Lin(-) cells) in vitro. MATERIALS AND METHODS: FR25Lin(-) cells were isolated by elutriation and cell sorting and cultured with a combination of cytokines for 7 days. Utilizing the membrane dye PKH-26, cultured cells were separated into two subsets based on their proliferation rates and assayed for progenitors and HSC. RESULTS: Fresh FR25Lin(-) cells were mostly quiescent; however, some of this population entered cell cycle after cytokine exposure reaching a peak 4 to 5 days after culture. Two subsets of cultured cells were isolated: 1) cells that had divided several times (PKH(dull) cells) and 2) cells that remained undivided or divided only once or twice (PKH(bright) cells). The PKH(dull) cells accounted for 94% of total viable cells in culture after 5 days. The PKH(dull) subset contained all the multi-potential in vivo progenitors (CFU S) and 10 times more committed progenitors (CFU-C). Quantitative analysis of HSC engraftment from the PKH(bright) subset demonstrated stem cell maintenance. For the PKH(dull) subset, on day 5, HSC numbers increased. By day 7, increased differentiation in the PKH(dull) population supports expanding differentiation divisions. CONCLUSIONS: Our primitive HSC population underwent different types of cell divisions stimulated by cytokines, resulting in subsets with different self renewal and differentiation potentials. This in vitro/in vivo model provides a useful tool for studies of early events during HSC self-renewal and differentiation. PMID- 12543108 TI - CD34+ cells from paroxysmal nocturnal hemoglobinuria (PNH) patients are deficient in surface expression of cellular prion protein (PrPc). AB - Cellular prion protein (PrP(c)) is a glycosylphosphatidylinositol (GPI)-anchored protein (GPI-AP) constitutively expressed by neurons but also in hematopoietic cells. In trasmissible spongiform encephalopathies, the protease-resistant form of prion (PrP (s c)) converts the host PrP(c) into the pathologic form. We have investigated PrP(c) expression in hematopoietic cells from paroxysmal nocturnal hemoglobinuria (PNH). In this disease, due to somatic mutations in PIG-A gene, biosynthesis of the (GPI)-anchor is impaired and affected cells lack membrane expression of all GPI-AP. Normal and PNH hematopoietic progenitors and paired wild-type (WT) and PIG-A mutant cell lines were used for analysis of intracellular and surface PrP(c) expression using flow cytometry and Western blot.By flow cytometry, PrP(c) was constitutively present on normal CD34(+) cells, including more immature CD38(dim) cells, as well as hematopoietic cell lines. Similar results were obtained in purified CD34(+). Phospholipase C treatment confirmed that PrP(c) was expressed on the membrane via the GPI-anchor. In PNH patients, GPI-AP-deficient CD34(+) cells lacked PrP(c) membrane expression. PIG-A-mutated cell lines (Jurkat, K562, C(EBV), A(EBV)), in contrast to their normal counterparts, did not express surface PrP(c). However, we detected intracellular PrP(c) at approximately equivalent levels in both normal and PIG-A-mutated cells using intracellular flow cytometry and Western blotting. Cells and cell lines with PNH phenotype together with their normal counterparts may be a suitable system to explore the function of membrane PrP(c) in the hematopoietic system. Conversely, PrP(c) is a good model to elucidate the fate of GPI-AP in PIG-A-deficient cells. PMID- 12543109 TI - Prognostic factors in allogeneic transplantation for patients with high-risk multiple myeloma after reduced intensity conditioning. AB - OBJECTIVES: The aim of this study was to identify prognostic factors for outcome of high-risk patients with multiple myeloma after allogeneic transplantation prepared by reduced intensity conditioning (RIC). MATERIALS AND METHODS: Data from 45 consecutive patients (median age 52 years, range 38-68), who received grafts from a sibling (n = 34) or unrelated donor (n = 11) were analyzed. Fourteen patients received an RIC allotransplant while chemosensitive (>/=partial remission [PR]), whereas 31 chemoresistant patients (0.05). SIGNIFICANCE: Results obtained do not support claims for surface flaw and/or defect healing through the formation of an SiOH layer at the surface of Duceram-LFC specimens over time. However, the polishing regime interrupted the defect population associated with flaws present in dry (control) samples of Duceram-LFC which can be attributed to an associated reduction of initial surface flaws and/or defects. PMID- 12543115 TI - Sublethal, 2-week exposures of dental material components alter TNF-alpha secretion of THP-1 monocytes. AB - OBJECTIVES: The aim of this study was to investigate the hypothesis that dental material components alter cytokine secretion from monocytes if applied for several weeks at sublethal doses. The current study significantly extended exposure times of monocytes to the components over times published in previous studies. These exposure times approached the estimated average life span of monocytes in the bloodstream. METHODS: Human THP-1 monocytes were exposed to 2 hydroxyethylmethacrylate (HEMA, 0-1.2mmol/l), triethyleneglycoldimethacrylate (TEGDMA, 0-0.75mmol/l), Hg(2+) (0-2 micromol/l), or Ni(2+) (0-20 micromol/l) for 2 weeks. The cells were then collected and additionally incubated for 24h, with or without bacterial lipopolysaccharide (LPS), a common component of dental plaque. TNF-alpha secretion from THP-1 was determined using by enzyme-linked immunosorbent assay. RESULTS: None of the dental material components induced TNF alpha from THP-1 by themselves, but LPS alone strongly induced TNF-alpha secretion as expected. HEMA and TEGDMA significantly suppressed (40-70%) TNF alpha secretion from cells stimulated with LPS. Hg(2+) at 2.0 micromol/l doubled TNF-alpha secretion from THP-1s stimulated with LPS over LPS alone. Ni(2+) did not significantly affect TNF-alpha secretion, with or without LPS exposure. Significance. The results in this study suggest that sublethal, 2-week exposures of some dental material components may alter TNF-alpha secretion from THP-1 monocytes when the cells are challenged. These alterations may influence the biological response of tissues to materials in an inflammatory intraoral environment. PMID- 12543116 TI - Reflection photoelasticity: a new method for studies of clinical mechanics in prosthetic dentistry. AB - OBJECTIVES: This research was conducted to demonstrate the effectiveness of reflective photoelasticity as an in vivo technique for monitoring the strain/stress distribution in prosthetic devices during function. METHODS: Seven subjects were selected for this study and divided into three groups according to the design of the prostheses. The buccal surfaces of the experimental prostheses were coated with a birefringent resin 2mm thick. Using a reflection polariscope, fringe patterns were documented on video tape during standardized in vivo loadings. Strain and stress levels (gamma(max) and tau(max)) were calculated from isochromatic fringe order determinations. The intra-observer variability in counting fringe orders and the accuracy level of reflective photoelasticity as compared to strain gauge results were also studied. The strain results of in vivo loading of the prostheses were analyzed with ANOVA and Scheffes tests. The intra observer variability was analyzed with the Friedman nonparametric test. Comparisons of photoelastic data and strain gauges were tested with Pearson's correlation. RESULTS: Coated areas accessible to normally incident light, produced comprehensive maps of strain distribution. Significant differences were found in the in vivo gamma(max) values for the prostheses designs (p<0.05). The test for intra-observer variability showed that no significant differences occurred in counting fringe orders. Strain values obtained with reflective photoelasticity showed a high correlation r=0.98 and 0.99) with values obtained at the same areas using strain gauges. SIGNIFICANCE: Reflective photoelasticity is a valid, reliable and accurate technique to be used for in vivo studies on the biomechanical behavior of prosthetic devices. PMID- 12543117 TI - A novel sensor for bite force determinations. AB - OBJECTIVES: The clinical usefulness, accuracy and precision of a novel bite force sensor based on force sensing resistors were tested in six subjects wearing maxillary removable partial dentures retained by conical crowns. METHODS: The surfaces of the sensor were manufactured in a silicone material that had mechanical properties similar to those of tough foodstuffs. In two separate series of standardized bite force tests, submaximum force levels were recorded with the sensor and with a strain gaged bite fork. Subjects were assisted in the loading tests with visual feedback instrumentation. Reliability estimates for the bite force sensor were calculated in order to show their reproducibility. Strain gages attached to the prostheses were used to determine the pattern of force distribution during loading tests. The bite force results obtained with the new bite force sensor and with the bite fork were analyzed with ANOVA and Scheffes tests. The strain patterns registered with strain gages were analyzed with F test. RESULTS: The bite force sensor and the bite fork transducer showed no statistically significant differences in respect of intra-individual bite force levels (range 50-300N). The bite forces registered with the new sensor were dependent on the loading position (p<0.05), sex (p<0.05) and test subject (p<0.05). The reliability of the new sensor for submaximum bite forces was calculated to be 93%. Strain gage results showed that the new sensor generated strain patterns of less variance (p<0.05) than the bite fork and therefore allowed for higher precision during biting tests. SIGNIFICANCE: The presented instrument has such clinical merits, as to favor its use in experimental clinical studies on the biomechanics of prosthetic appliances. PMID- 12543118 TI - On the accuracy of some in vitro models for mechanical studies of maxillary removable partial dentures. AB - OBJECTIVES: Comparative in vitro/in vivo studies show wide differences in functional strain magnitudes and patterns of functional strain distribution in prosthetic devices. The aim of this study was to evaluate the accuracy of three in vitro models for biomechanical studies of prosthetic devices. METHODS: Strain gages were attached to the test RPDs of six subjects. In vitro maxillary models with simple (model A), intermediate (model B) and advanced (model C) features for the anatomical simulation of supporting structures were manufactured for each subject. The dentures were subjected to two separate series of standardized functional and laboratorial loading tests. The principal maximum strain value (epsilon (1)) obtained for each gage and experiment was used to calculate the intra-experiment variance and inter-experiment variance for the in vivo and in vitro experiments. The integrated strain gage difference value between the in vitro and in vivo data (Sigma(diff)) was calculated for every loading test and compared to the in vivo inter-experiment variance with ANOVA and Scheffes test. RESULTS: The bite forces recorded were similar for each subject and loading position and between the two loading sessions. The inter- and intra-experiment variance was found to be higher for in vivo loadings than for in vitro. Results showed that the (Sigma(diff)) values for model B and model C were significantly different from the in vivo strain inter-experiment variance (p<0.05). SIGNIFICANCE: This study showed that the anatomical simulation of in vitro models is insufficient to allow for accurate mechanical analyses of maxillary RPDs and that only simple verifications of the strain levels in prosthetic appliances can be attained in vitro. PMID- 12543119 TI - Optimizing filler content in an adhesive system containing pre-reacted glass ionomer fillers. AB - OBJECTIVES: (1) To evaluate the optimal concentration of the calcium type pre reacted glass-ionomer (PRG-Ca) fillers in experimental light-activated adhesives for bonding to tooth substrates and fluoride release; (2) to evaluate the polymerization reactivity of these experimental filled adhesives; and (3) to examine the ultrastructure of the resin-dentin interface bonded with the commercial version of this self-etching adhesive (Imperva Fluoro Bond, Shofu Inc., Kyoto, Japan). MATERIALS AND METHODS: PRG-Ca fillers were prepared from the acid-base reaction of calcium type fluoroaluminosilicate glass with poly(acrylic acid) in water. Different experimental light-cured adhesives were prepared, with 0-57wt% PRG-Ca fillers and 3wt% aerosil silica incorporated in a resin matrix. Twenty-four hours shear bond strengths to bovine enamel and dentin were assessed using the adhesive with/or without accelerated aging. The cumulative amount of fluoride released from the cured adhesives and differential scanning calorimetry (DSC) measurements were also conducted. 180-grit SiC paper polished human dentin disks were treated with Imperva Fluoro Bond that contains 17wt% PRG-Ca fillers and prepared for TEM examination. RESULTS: Increase in PRG-Ca filler content of experimental adhesives was highly correlated with the decrease in bond strengths to dentin (r=-0.96 to -0.99), the increase in cumulative fluoride release, (r=0.97) and the decrease in polymerization energy (cal/g) of the adhesives (r= 0.99) (p<0.001). A PRG-Ca content of 17wt% provided optimal bond strength to enamel and dentin after accelerated aging and fluoride release. TEM showed that salt-like matrices were present around the hydrogels and remnant glass cores of the PRG-Ca fillers. The commercial self-etching primer completely dissolved the smear layer and formed 2 micrometer thick hybrid layers in intact dentin. SIGNIFICANCE: The use of silanized PRG-Ca fillers provides good adhesion and a source of fluoride release in a commercially available self-etching adhesive system. PMID- 12543120 TI - Effect of slow-curing on cavity wall adaptation using a new intensity-changeable light source. AB - OBJECTIVE: The purpose of the study was to investigate the effects of a new intensity-changeable light source Curetron 7 (CT-7) devised for the slow-curing on cavity wall adaptation in the adhesive composite restorations, as well as the microhardness of the cured composite. METHODS: Microhardness of both top and bottom surfaces was measured by an indentation method for 2mm thick cylindrical specimens, and cavity adaptation was evaluated in cylindrical dentin cavities (slashed circle 3.5x1.5mm(2)) of human extracted molars bonded to a hybrid resin composite. The irradiation was done with CT-7, VIP or Candelux (CDX) under the following five conditions: CT1, 600mW/cm(2)x30s (CT-7); CT2, 230mW/cm(2)x20s+600mW/cm(2)x20s (CT-7); CT3, 230mW/cm(2)x20s+pause x 10s+600mW/cm(2)x20s (CT-7); VIP, 300mW/cm(2)x3s+pause x 3min+600mW/cm(2)x30s (VIP); CDX, 200mW/cm(2)x10s+600mW/cm(2)x30s. RESULTS: Five irradiation conditions had no influence on the microhardness for either top or bottom surface (Scheffe, p>0.05). In conditions CT3, VIP and CDX, top surfaces were harder than bottom surfaces (Student's t-test, p<0.05). The statistical analysis revealed no difference in adaptation among the locations along the cavity walls for each irradiation conditions (Kruskal-Wallis, p>0.05). In comparison of poled data for each condition, the best adaptation was seen in condition CT3, and the second in condition VIP (Kruskal-Wallis, p<0.05). SIGNIFICANCE: These results proved the efficacy of the slow-curing method combined with the interval between two irradiations with low intensity and high intensity. CT-7 could be useful for the adhesive composite restorations. PMID- 12543121 TI - Phosphoric acid and various transition metal salt solutions as a combined etchant and activator prior to the use of an anaerobic adhesive. AB - OBJECTIVES: The objectives of this experiment were to determine whether various transition metal salts in 37% o-phosphoric acid could both activate and etch an enamel surface prior to the use of an anaerobic adhesive. METHOD: Stainless steel attachments were bonded to human enamel using an anaerobic adhesive. In each case, the enamel was etched and activated using a solution of 37% o-phosphoric acid containing various transition metal sulfates and chlorides. After bench curing, the specimens were shear bond tested to failure and the load at debond recorded in each case. RESULTS: The results were analyzed using mean force to debond (N) and 95% confidence intervals. Kaplan-Meier survival probabilities and log-rank tests were also performed.Conclusions. Under the conditions of this experiment, the sulfate and chloride of copper in acid were the most effective etching/activating solutions. There was no significant difference in the mean force to debond between the copper (II) sulfate and copper (II) chloride. The chloride in acid was, however, the only one of the two to produce a conventional etch pattern on the surface of the enamel. SIGNIFICANCE: It is possible to render the enamel surface both retentive and active towards anaerobic adhesives, such that relatively unreactive substrates can be bonded to enamel using such adhesives. PMID- 12543122 TI - Enhanced antibacterial activity in Hydra polyps lacking nerve cells. AB - The nervous system evolved within cnidarians. When assessing antibacterial activity in the freshwater polyp Hydra, we observed a strong correlation between the number of neurons present and the antibacterial activity. Tissue lacking neurons had a drastically enhanced antibacterial activity against Gram-positive (Bacillus subtilis) and Gram-negative (E. coli) bacteria compared to control tissue. The results indicate direct and strong neural influences on immunity in the phylogenetically oldest animals having a nervous system. PMID- 12543123 TI - Emergence and evolution of functional heavy-chain antibodies in Camelidae. AB - Antibodies of jawed-vertebrates are composed of paired heavy (H) and light (L) polypeptide chains. Surprisingly, the sera of camelids, nurse shark and wobbegong shark, and possibly ratfish contain antibodies that lack L-chains. In camelids, these Heavy-chain antibodies (HCAbs) are gamma-isotypes, and are functional in antigen binding. In this review we focus on the dedicated immunoglobulin (Ig) genes that encode the HCAb in Camelidae (camels, dromedaries and llamas), about their origin, and how these camel immunoglobulins evolved and acquire a large and diverse repertoire of antigen binding sites in absence of the H-L combinatorial diversity. PMID- 12543124 TI - Lipopolysaccharides induce intestinal serum amyloid A expression in the sea cucumber Holothuria glaberrima. AB - We have previously characterized the first invertebrate homolog of serum amyloid A (SAA) proteins in the sea cucumber Holothuria glaberrima, where its expression is associated with intestinal regeneration, suggesting a possible involvement of SAA proteins in intestinal morphogenesis. Here we show that bacterial lipopolysaccharides (LPS) trigger a coelomocyte-mediated immune response in H. glaberrima, inducing an approximately threefold increase in coelomocyte phagocytic activity. Furthermore, LPS induces an approximately fourfold increase in SAA mRNA levels in non-regenerating intestines. These results show that in H. glaberrima, LPS act as an immune activator and that SAA expression can be modulated by immune-associated processes. PMID- 12543125 TI - Cloning and expression analysis of rainbow trout Oncorhynchus mykiss interferon regulatory factor 1 and 2 (IRF-1 and IRF-2). AB - Two rainbow trout (Oncorhynchus mykiss) genes for interferon regulatory factors (IRF) 1 and 2 have been cloned and sequenced. The IRF-1 cDNA contains an open reading frame (ORF) of 996 nucleotides that translates into a 331 amino-acid putative peptide, with a 5' untranslated region (UTR) of 145bp and a 3' UTR of 481bp. The IRF-2 cDNA contains a 1035bp ORF that translates into a 344 amino-acid putative peptide, with a 5' UTR of 146bp and a 3' UTR of 925bp. In vivo, IRF-1 and IRF-2 are constitutively expressed in head kidney, gill and spleen but not liver. Both genes were induced in all the tissues examined. IRF-1 but not IRF-2 expression was significantly increased at the site of injection 1 week after DNA vaccination against viral haemorrhagic septicaemia virus. In vitro, IRF-1 and IRF 2 transcripts are present in unstimulated rainbow trout gonad cells and are up regulated by poly I/C. PMID- 12543126 TI - Molecular cloning and characterisation of a carp (Cyprinus carpio) cytokine-like cDNA that shares sequence similarity with IL-6 subfamily cytokines CNTF, OSM and LIF. AB - In the course of suppression subtractive hybridisation between sodium alginate induced peritoneal cells (SA-PC) and normal head kidney cDNAs in common carp (Cyprinus carpio), a cytokine-like cDNA clone was found. The clone, named M17, contains a 1600bp nucleotide sequence that encodes a 215 amino acid putative protein that would have a pI of 9.01 and would include a 33 amino acid signal peptide. The 3' untranslated region has seven ATTTA mRNA destabilising motifs that are common in cytokines and oncogenes. In a BLASTP search, M17 was most similar to chicken ciliary neurotrophic factor (CNTF) with 25% amino acid identity, followed by mammalian CNTF, cardiotrophin-1 and leukemia inhibitory factor (LIF) all of which belong to the IL-6 subfamily. However, M17 has some differences with CNTF in that CNTF has no signal sequence, the gene organisation of M17 is three exons and two introns, whereas that of CNTF is two exons and one intron, M17 has seven cysteines while CNTF has one cysteine, and M17 mRNA is detected in peripheral blood leukocytes as well as brain, whereas CNTF is expressed only in the nervous system. Compared to other members in the IL-6 subfamily cytokines, M17's cysteine positions and gene organisation are similar to those of oncostatin M and LIF, although amino acid identities are only 15-17%. Southern hybridisation suggested that M17 is a single copy gene. SA-PC showed significantly higher M17 mRNA levels than normal head kidney cells, which are considered to be a source of the SA-PC, indicating that M17 is inducible by inflammatory stimulation. PMID- 12543127 TI - Interleukin-1beta does not contribute to genetic strain-based differences in iNOS expression and activity in chicken macrophages. AB - The expression of IL-1beta and inducible nitric oxide synthase (iNOS) from iNOS hypo (GB2, B(6)B(6)) and hyper (K-strain, B(15)B(15)) responder chickens was examined. Compared to GB2, macrophages from K-strain expressed higher iNOS mRNA as quantitated by reverse transcriptase polymerase (RT-PCR) chain reaction after stimulation with 1 microgram/ml of Escherichia coli (E. coli) lipopolysaccharide (LPS). On the contrary, IL-1beta mRNA expression was comparable between K and GB2 macrophages at 3h post-LPS stimulation but persisted up to 9h only in GB2 macrophages. The LPS-inducible interleukin-1 (IL-1) surface receptor expression, measured by flow cytometry, was higher in GB2 than on K-strain macrophages. Blocking of IL-1 receptor by the anti-IL-1 receptor antibody reduced the LPS mediated iNOS expression by 50% as quantified by competitive RT-PCR. Furthermore, iNOS activity (nitrite) was also reduced to 50%. However, this magnitude of inhibition was similar in both K and GB2 macrophages. While these observations suggest that IL-1beta is involved in mediating LPS-induced iNOS expression and activity, the differential response of GB1 and K-strain macrophages in terms of LPS-induced iNOS expression and activity is unlikely to be modulated by IL-1beta. PMID- 12543128 TI - Establishment of immune competence in the avian GALT during the immediate post hatch period. AB - Population dynamics of intestinal lymphocytes and the temporal development of lymphocyte functions were studied in broiler chicks during the first 2 weeks post hatch. This period is of major immunological importance as the chick is immediately exposed to environmental antigens and pathogens. We show that the gut associated lymphoid tissue contains functionally immature T and B lymphocytes at hatch, and that function is attained during the first 2 weeks of life as demonstrated by mRNA expression of both ChIL-2 and ChIFNgamma. Functional maturation occurred in two stages: the first-during the first week post-hatch, and the second during the second week, which was also accompanied by an increase in lymphocyte population, as determined by expression of antigen receptor genes. Evidence is presented to show that in the intestinal milieu cellular immune responses mature earlier, and are a prerequisite for humoral responses. Hence, the lack of antibody response in young chicks is primarily due to immaturity of T lymphocytes. PMID- 12543129 TI - Immunobiology of chicken germinal center: I. Changes in surface Ig class expression in the chicken splenic germinal center after antigenic stimulation. AB - The germinal center (GC) develops after antigenic stimulation and is thought to occur at the site of various immune responses. We separated a single GC from chicken spleen after antigenic stimulation. Flow cytometric analysis of the cells derived from a single GC and RT-PCR analysis of Ig mRNA expression in GC was performed. Direct evidence indicates that: (1) there was a considerable difference in the cell population of each GC, (2) the ratio of CD3(+) cells in a GC remains constant at 10-20%, (3) the highest proportion of sIgY(+) cells in a GC occurs 1 week after the time of highest proportion of sIgM(+) cells, and (4) RT-PCR analysis was used to detect IgY mRNA expression in a GC. The continuous existence of CD3(+) cells, the alterations in sIgM(+) and sIgY(+) cell ratios, and the expression of IgY mRNA strongly suggest that Ig class switching occurs in the GC during an immune response. PMID- 12543131 TI - Computer model for recognition of functional transcription start sites in RNA polymerase II promoters of vertebrates. AB - This paper introduces a new computer system for recognition of functional transcription start sites (TSSs) in RNA polymerase II promoter regions of vertebrates. This system allows scanning complete vertebrate genomes for promoters with significantly reduced number of false positive predictions. It can be used in the context of gene finding through its recognition of the 5' end of genes. The implemented recognition model uses a composite-hierarchical approach, artificial intelligence, statistics, and signal processing techniques. It also exploits the separation of promoter sequences into those that are C+G-rich or C+G poor. The system was evaluated on a large and diverse human sequence-set and exhibited several times higher accuracy than several publicly available TSS finding programs. Results obtained using human chromosome 22 data showed even greater specificity than the evaluation set results. The system has been implemented in the Dragon Promoter Finder package, which can be accessed at http://sdmc.krdl.org.sg:8080/promoter/. PMID- 12543132 TI - MolSpace: a computer desktop tool for visualization of massive molecular data. AB - The authors have developed a software tool, MolSpace, to visualize massive molecular datasets. MolSpace can project a set of massive multivariate data onto a visual space (two- or three-dimensional space) by means of principal component analysis. MolSpace allows users not only to draw a scatter diagram of the data but also to display their two- or three-dimensional molecular structures as the objects in that space. With a probe (a molecular object) the user can navigate vast data spaces, thus facilitating understanding of the data structure. In addition, partial space searching is also available that is based on similarity searching techniques. It is possible to interrogate a three-dimensional structure of a chemical compound that corresponds to each object on the space in real time. The detail of the system is discussed with an illustrative example. PMID- 12543133 TI - Designing supramolecular structures from models of cyclic peptide scaffolds with heterocyclic constraints. AB - Cyclic peptides containing oxazole and thiazole heterocycles have been examined for their capacity to be used as scaffolds in larger, more complex, protein-like structures. Both the macrocyclic scaffolds and the supramolecular structures derived therefrom have been visualised by molecular modelling techniques. These molecules are too symmetrical to examine structurally by NMR spectroscopy. The cyclic hexapeptide ([Aaa-Thz](3), [Aaa-Oxz](3)) and cyclic octapeptide ([Aaa Thz](4), [Aaa-Oxz](4)) analogues are composed of dipeptide surrogates (Aaa: amino acid, Thz: thiazole, Oxz: oxazole) derived from intramolecular condensation of cysteine or serine/threonine side chains in dipeptides like Aaa-Cys, Aaa-Ser and Aaa-Thr. The five-membered heterocyclic rings, like thiazole, oxazole and reduced analogues like thiazoline, thiazolidine and oxazoline have profound influences on the structures and bioactivities of cyclic peptides derived therefrom. This work suggests that such constrained cyclic peptides can be used as scaffolds to create a range of novel protein-like supramolecular structures (e.g. cylinders, troughs, cones, multi-loop structures, helix bundles) that are comparable in size, shape and composition to bioactive surfaces of proteins. They may therefore represent interesting starting points for the design of novel artificial proteins and artificial enzymes. PMID- 12543134 TI - Formation of neutral molecules of potential stellar interest by neutralisation of negative ions in a mass spectrometer. The application of experiment and molecular modelling in concert. AB - This paper is a modified version of a lecture which describes the synthesis, structure and reactivity of some neutral molecules of stellar significance. The neutrals are formed in the collision cell of a mass spectrometer following vertical Franck-Condon one electron oxidation of anions of known bond connectivity. Neutrals are characterised by conversion to positive ions and by extensive theoretical studies at the CCSD(T)/aug-cc-pVDZ//B3LYP/6-31G(d) level of theory. Four systems are considered in detail, viz (i) the formation of linear C(4) and its conversion to the rhombus C(4), (ii) linear C(5) and the atom scrambling of this system when energised, (iii) the stable cumulene oxide CCCCCO, and (iv) the elusive species O(2)C-CO. This paper is not intended to be a review of interstellar chemistry: examples are selected from our own work in this area. PMID- 12543135 TI - Novel pharmacophore-based methods reveal gossypol as a reverse transcriptase inhibitor. AB - In a program to identify new structural entities for the inhibition of the HIV-1 reverse transcriptase (RT) enzyme via database searching, a series of RT pharmacophores were developed. By utilising a novel filtering technique, the National Cancer Institute database of compounds was scanned producing 15 compounds to be screened for activity. A notable inclusion was a series of gossypol derivatives. The testing of a series of compounds revealed the parent compound gossypol to be an HIV-1 reverse transcriptase inhibitor. These results suggest that at least a part of its anti-HIV activity is due to gossypol targeting the non-nucleoside inhibitor binding pocket of RT. PMID- 12543136 TI - Discrimination and selection of new potential antibacterial compounds using simple topological descriptors. AB - The aim of the work was to discriminate between antibacterial and non antibacterial drugs by topological methods and to select new potential antibacterial agents from among new structures. The method used for antibacterial activity selection was a linear discriminant analysis (LDA). It is possible to obtain a QSAR interpretation of the information contained in the discriminant function. We make use of the pharmacological distribution diagrams (PDDs) as a visualizing technique for the identification and selection of new antibacterial agents. PMID- 12543137 TI - Prediction of dihydrofolate reductase inhibition and selectivity using computational neural networks and linear discriminant analysis. AB - A data set of 345 dihydrofolate reductase inhibitors was used to build QSAR models that correlate chemical structure and inhibition potency for three types of dihydrofolate reductase (DHFR): rat liver (rl), Pneumocystis carinii (pc), and Toxoplasma gondii (tg). Quantitative models were built using subsets of molecular structure descriptors being analyzed by computational neural networks. Neural network models were able to accurately predict log IC(50) values for the three types of DHFR to within +/-0.65 log units (data sets ranged approximately 5.5 log units) of the experimentally determined values. Classification models were also constructed using linear discriminant analysis to identify compounds as selective or nonselective inhibitors of bacterial DHFR (pcDHFR and tgDHFR) relative to mammalian DHFR (rlDHFR). A leave-N-out training procedure was used to add robustness to the models and to prove that consistent results could be obtained using different training and prediction set splits. The best linear discriminant analysis (LDA) models were able to correctly predict DHFR selectivity for approximately 70% of the external prediction set compounds. A set of new nitrogen and oxygen-specific descriptors were developed especially for this data set to better encode structural features, which are believed to directly influence DHFR inhibition and selectivity. PMID- 12543138 TI - Comparison of chemical clustering methods using graph- and fingerprint-based similarity measures. AB - This paper compares several published methods for clustering chemical structures, using both graph- and fingerprint-based similarity measures. The clusterings from each method were compared to determine the degree of cluster overlap. Each method was also evaluated on how well it grouped structures into clusters possessing a non-trivial substructural commonality. The methods which employ adjustable parameters were tested to determine the stability of each parameter for datasets of varying size and composition. Our experiments suggest that both graph- and fingerprint-based similarity measures can be used effectively for generating chemical clusterings; it is also suggested that the CAST and Yin-Chen methods, suggested recently for the clustering of gene expression patterns, may also prove effective for the clustering of 2D chemical structures. PMID- 12543139 TI - A priori molecular descriptors in QSAR: a case of HIV-1 protease inhibitors. I. The chemometric approach. AB - A quantitative structure-activity relationship (QSAR) study on 48 peptidic HIV-1 protease inhibitors was performed. Fourteen a priori molecular descriptors were used to build QSAR models. Hierarchical cluster analysis (HCA), principal component analysis (PCA) and partial least squares (PLS) regression were employed. PLS models with 32/16 (model I) and 48/0 (model II) molecules in the training/external validation set were constructed. The a priori molecular descriptors were related to two energetic variables using PLS. HCA and PCA on data from model II classified the inhibitors as slightly, moderately and highly active; three principal components, the chemical nature of which has been highlighted, are enough to describe the enzyme-inhibitor binding. Model I (r(2)=0.91, q(2)=0.84) is comparable to literature models obtained by various QSAR softwares, which justified the use of a priori descriptors. PMID- 12543140 TI - Assessing the performance of OMEGA with respect to retrieving bioactive conformations. AB - OMEGA is a rule-based program which rapidly generates conformational ensembles of small molecules. We have varied the parameters which control the nature of the ensembles generated by OMEGA in a statistical fashion (D-optimal) with the aim of increasing the probability of generating bioactive conformations. Thirty-six drug like ligands from different ligand-protein complexes determined by high resolution (< or =2.0A) X-ray crystallography have been analyzed. Statistically significant models (Q(2)> or =0.75) confirm that one can increase the performance of OMEGA by modifying the parameters. Twenty-eight of the bioactive conformations were retrieved when using a low-energy cut-off (5 kcal/mol), a low RMSD value (0.6A) for duplicate removal, and a maximum of 1000 output conformations. All of those that were not retrieved had eight or more rotatable bonds. The duplicate removal parameter was found to have the largest impact on retrieval of bioactive conformations, and the maximum number of conformations also affected the results considerably. The input conformation was found to influence the results largely because certain bond angles can prevent the bioactive conformation from being generated as a low-energy conformation. Pre-optimizing the input structures with MMFF94s improved the results significantly. We also investigated the performance of OMEGA in connection with database searching. The shape-matching program Rapid Overlay of Chemical Structures (ROCS) was used as search tool. Two multi conformational databases were built from the MDDR database plus the 36 compounds; one large (maximum 1000 conformations/mol) and one small (maximum 100 conformations/mol). Both databases provided satisfactory results in terms of retrieval. ROCS was able to rank 35 out of 36 X-ray structures among the top 500 hits from the large database. PMID- 12543141 TI - A new method for mapping macromolecular topography. AB - A new method, using circular variance, is introduced for mapping macromolecular topography. Circular variance, generally used to measures angular spread, can be used to characterize of molecular structures based on a simple idea. It will be shown that the circular variance of vectors drawn from some origin to a set of points is well correlated with the degree to which that origin is inside/outside the chosen points. In addition, it has continuous derivatives that are also easy to compute. This concept will be shown to be useful for: (i) distinguishing between atoms near the surface of a macromolecule and those in either the deep interior or remote exterior; (ii) identifying invaginations (even shallow ones); and (iii) detecting linker regions that interconnect two domains. PMID- 12543142 TI - Transmission-blocking vaccine of vivax malaria. AB - Malaria remains one of the leading causes of both morbidity and mortality of humans residing in tropical countries. For many malarious regions outside of Africa, development of effective transmission-blocking vaccines will require coverage against both Plasmodium falciparum and Plasmodium vivax. The genes coding for two potential P. vivax transmission-blocking antigens, Pvs25 and Pvs28, have been cloned. Mice vaccinated with yeast-produced recombinant proteins Pvs25 and Pvs28 adsorbed to aluminum hydroxide developed strong antibody responses against the immunogens. The development of oocysts in mosquitoes was completely inhibited when these antisera were ingested with the P. vivax Salvador (Sal) I strain-infected chimpanzee blood. In a large collection of P. vivax field isolates, we found only 5 nucleotide changes that would result in amino acid substitutions in Pvs25. In contrast, the Pvs28 gene had 22 nucleotide changes that would result in conservative amino acid substitutions. How the antigenic polymorphism of Pvs25 and Pvs28 would affect the efficacy of Sal I based vaccine remains to be elucidated. Clinical trials with Pvs25 and the P. falciparum ortholog Pfs25 are in preparation. PMID- 12543143 TI - Analysis of Wuchereria bancrofti infections in a village community in northern Nigeria: increased prevalence in individuals infected with Onchocerca volvulus. AB - Infections with Wuchereria bancrofti causing lymphatic filariasis still represent one of the major health problems in the tropics, with 120 million people infected and over 750 million exposed to this filarial parasite. We have studied lymphatic filariasis infections as part of a multi-parasite survey in a village community in the savannah of northern Nigeria. We analysed serum samples from 341 individuals aged 5-70 years, detecting a W. bancrofti circulating antigen using the commercially available ICT Filariasis card test. The prevalence of infections was 10% and clearly age-dependent, increasing from below 2% in children to over 20% in subjects older than 40 years. Measuring IgG4 antibodies against the recombinant W. bancrofti antigen SXP1 showed that 36% of all tested individuals had been at least exposed to the parasite. Antibody levels also increased very significantly with age. A further analysis measuring Onchocerca volvulus-specific IgG4 antibodies showed a very significant association between infections with O. volvulus and those with W. bancrofti. Our data show that infections with W. bancrofti in Nigeria are still a frequently occurring health problem, since they are more prevalent than previously reported, and that individuals with an O. volvulus infection are more often infected with W. bancrofti than expected statistically. PMID- 12543144 TI - Trematode fauna of prosobranch snails of the genus Semisulcospira in Lake Biwa and the connected drainage system. AB - The parasite fauna of prosobranch snails of the genus Semisulcospira was surveyed in Lake Biwa and the adjacent water system. One aspidogastrean and 28 digenetic trematode taxa were detected in 19209 snails consisting of 10 morphological species. There was no trematode species peculiar to members of the subgenus Biwamelania that is endemic to the Lake Biwa water system. However, one species, Notocotylus magniovatus, was found only in the non-endemic subgenus Semisulcospira. Of 23 digenean taxa detected in more than one host, 13 were distributed in both the lake and the tributaries. Seven of these had host taxa, more than 1% of which were infected with the parasite in both the lake and the tributaries, four had such hosts only in the tributaries, and two had no such hosts. Three species detected only in Lake Biwa were previously reported from other rivers in Japan. In the seven species detected only in the tributaries, two species had life cycles that could be maintained only in rivers. These results indicate that the core areas for the distribution of parasites of Semisulcospira are tributaries, and the lake is a sink for these species. These results contradict the expectation that the parasite fauna should be richer in the lake than in tributaries because the lake is a stable habitat over a geological time scale and has more divergent freshwater animals than the adjacent water system. PMID- 12543145 TI - Adult worms of the rodent intestinal nematode, Strongyloides venezuelensis, successfully invade chick intestinal mucosa. AB - In order to study the mucosal invasion of a rodent intestinal nematode in bird intestine, chicks were infected with the intestinal nematode of rodents, Strongyloides venezuelensis, by subcutaneous larva inoculation and adult worm implantation. No evidence was obtained for larvae reaching the lungs or the intestine after infective larva inoculation. Adult worms implanted in the small intestine invaded the mucosa and remained there at least for 24 h, whereas those implanted in the caecum were trapped by mucus, and did not invade the mucosa. Mucosal invasion of adult worms in the small intestine was confirmed by histological examination. The number of adult worms in the intestinal mucosal tissue dropped rapidly within the first 24 h, which was associated with infiltrating granulocytes around the worms. The present study suggests that S. venezuelensis adult worms are able to invade the intestinal tissue of chicks, which do not belong to the vertebrate class of its normal definitive host, but that they are eliminated rapidly by mucosal defense system of the bird. PMID- 12543146 TI - Characterisation of a sexual stage-specific gene encoding ORC1 homologue in the human malaria parasite Plasmodium falciparum. AB - The origin recognition complex (ORC) is a multisubunit protein composed of six polypeptides that binds to replication origins and is essential for the initiation of chromosomal DNA replication. Using the Vectorette technique, we have isolated a novel gene encoding an ORC1-like protein from the human malaria parasite Plasmodium falciparum. The gene has no introns and encodes a protein (PfORC1) of 1189 amino acid residues with a predicted molecular mass of 139 kDa. PfORC1 contains all conserved sequences in the ORC1/Cdc6/Cdc18 family and displays the highest homology to the Schizosaccharomyces pombe ORC1. However, PfORC1 possesses an extensive N-terminal segment with several interesting features including multiple potential phosphorylation sites, a large proportion of charged amino acids, four copies of a heptamer repeat, two nuclear localisation signals, and a leucine zipper motif. Southern blot analyses show that the Pforc1 gene is present as a single copy per haploid genome and is located on chromosome 12. A 5600 nucleotide transcript of this gene is expressed predominantly in the sexual erythrocytic stage, indicating that PfORC1 may be involved in gametogenesis during which DNA is quickly replicated. PMID- 12543147 TI - Oral artesunate prevents Plasmodium berghei Anka infection in mice. AB - Artesunate, a semi-synthetic derivative of a naturally occurring anti-malarial artemisinin was compared with chloroquine in C57BL/6 mice infected with Plasmodium berghei Anka (PbA). A 7-day oral administration of artesunate prevented parasitaemia at 10 mg/kg/day. However, recrudescence of parasitaemia and cerebral malaria occurred upon cessation of treatment followed by death within 28 days. However, a 14-day course of artesunate (100 mg/kg/day) prevented completely the development of parasitaemia and cerebral malaria with a survival of more than 60-days as did 10 mg/kg/day chloroquine. These data demonstrate that oral artesunate inhibits PbA and prevents cerebral malaria, but needs to be administered at high dose and for prolonged time to eradicate PbA infection in mice. PMID- 12543148 TI - Expansion of unconventional T cells with natural killer markers in malaria patients. AB - Immunological states during human malarial infection were examined. In parallel with parasitemia and anemia, granulocytosis was induced in the blood of patients, especially those infected with Plasmodium (P.) falciparum. At that time, the level of lymphocytes remained unchanged or slightly increased in the blood. However, the distribution of lymphocyte subsets was modulated, showing that the proportion of CD56(+)T cells, CD57(+)T cells, and gammadeltaT cells (i.e. all unconventional T cells) had increased in patients infected with P. falciparum or P. vivax. This phenomenon occurred at the early phase of infection and disappeared in the course of recovery. The data from patients with multiple attacks of P. vivax infection showed that there was no augmentation of these responses. In adult cases, the increase in the proportion of unconventional T cells seemed to closely parallel disease severity. However, all these responses were weak in children, even those infected with P. falciparum. In conjunction with accumulating evidence from mouse malaria experiments, the present results suggest that the immunological state induced by malarial infection might mainly be an event of unconventional T cells and that the immunological memory might not be long-lasting, possibly due to the properties of unconventional T cells. PMID- 12543149 TI - Intraepithelial infiltration of eosinophils and their contribution to the elimination of adult intestinal nematode, Strongyloides venezuelensis in mice. AB - Eosinophils were examined for the capacity of attacking Strongyloides venezuelensis adult worms in the intestinal mucosa by using interleukin (IL)-5 transgenic mice. In IL-5 transgenic mice, most of the subcutaneously inoculated infective larvae were killed during migration, and only a few worms could reach the small intestine. When the same number of adult worms were surgically implanted in the small intestine of IL-5 transgenic and control mice, fecal egg output as well as the number of adult worms recovered from the intestine was significantly lower in IL-5 transgenic mice. In the intestinal mucosa of IL-5 transgenic mice, large number of eosinophils was present in the lamina propria even before adult worm implantation. The number of eosinophils increased significantly as early as 24 h after implantation and tripled by day 3, whereas mucosal eosinophilia remained low in wild-type mice. Most notably, eosinophils infiltrated into the intestinal epithelium and surrounded adult worms in IL-5 transgenic mice, which was never seen in wild-type control mice. However, IL-5 transgenic mice required the same period as normal mice to completely expel implanted adult worms. The amount of specific IgA as well as total IgA in the stool was high in IL-5 transgenic mice before adult worm implantation, and dropped rapidly after adult worm implantation. The present study suggests that eosinophils are capable of attacking adult nematodes in the intestinal epithelia, probably in conjunction with secretory IgA, although they are not enough for the complete worm expulsion. PMID- 12543150 TI - A mathematical model for the transmission of Plasmodium vivax malaria. AB - We have proposed a mathematical model for the transmission of Plasmodium vivax malaria quantitatively, which is adjusted to the infected region, Guadalcanal, in the Solomon Islands. The simulation of a transmission model will be instrumental in planning the malaria control strategy. A characteristic of the life cycle of P. vivax is that a sporozoite injected into the blood stream by a mosquito bite may sometimes stay in a hepatocyte as a hypnozoite. Therefore, we have incorporated a phenomenon of renewed infections caused by a relapse into the transmission model. Also through the simulations we have attempted to evaluate the decline in prevalence caused by the programs of selective mass drug administration (MDA) and vector control such as the distribution of permethrin treated bednets. The simulations have indicated that the concentrated repetition of MDA at 1-week intervals would reduce the prevalence of vivax malaria swiftly in the beginning and would keep the parasite rate below 1% for a few years but the prevalence would increase thereafter. In contrast, the parasite rate would remain below 1% for a long time if a trial of 1 or 2 times MDA is accompanied with some reduction of the vectorial capacity by the enforcement of vector control. In any case, it is important to beware of relapse cases because even after the execution of MDA it takes a long time to decrease the proportion of hypnozoite carriers. PMID- 12543152 TI - Adaptive plasticity in the otolith-ocular reflex. AB - This review focuses on the plasticity in the vestibulo-ocular reflex (VOR), especially in the otolith-ocular reflex (OOR). The VOR is a mechanism for the production of rapid compensatory eye movements during head movements. The VOR is under adaptive control which corrects VOR performance when visual-vestibular mismatch arises during head movements. It has been demonstrated that chronic exposure to certain visual environments, those generated by magnifying lenses and reversing prisms, alter the gain and phase of VOR in the dark in numerous species. Most experiments concerning such modification of the VOR have used semicircular canal stimulation. The VOR consists of the semicircular-ocular reflex (ScOR) and the OOR. There are few results regarding the relationship between the gain of the ScOR and the OOR. This review summarizes the studies on plasticity in the OOR. In addition, the difficulty in evaluating the OOR using current conventional methods including earth horizontal axis (EHA) rotation, off vertical axis rotation (OVAR) and linear sled, is discussed. We believe that the ScOR and the OOR share common neural pathways in such a way that a change in the synaptic efficacy of one pathway is accompanied by a change in the other. PMID- 12543153 TI - Central tinnitus. AB - Tinnitus is likely initiated by a discontinuity in the spontaneous or low-level stimulus induced neural activity across auditory nerve fibers with different characteristic frequency (CF). This discontinuity may be caused by functional loss of outer hair cells in those regions where inner hair cells are preserved. The reduced spontaneous activity for nerve fibers with CFs in the hearing loss range may result in a reduction of lateral inhibition at more central levels. This reduced lateral inhibition of neurons with CFs close to the edge frequency of the audiogram induces hypersensitivity and hyperactivity in these neurons. Persistent changes in lateral inhibition result in increased numbers of neurons that are tuned to a limited range of frequencies at the edge of the cochlear lesion. Thus, the frequency map in auditory cortex, the tonotopic map, becomes reorganized as it reflects these changes. The spontaneous neuronal firings in the auditory cortex after insults that cause tinnitus show increased synchrony, thereby mimicking one aspect of the responses to normal sound stimulation. All long-standing tinnitus may thus be called central tinnitus, despite the fact that it is initiated by cochlear hearing loss or is localized to the hearing loss ear. PMID- 12543154 TI - Wallenberg's syndrome: neurotological classification. AB - OBJECTIVES: Visually induced eye movements were investigated in 26 patients with Wallenberg's syndrome. METHODS: Slow-phase optokinetic nystagmus (OKN) velocities, pursuit gains, and percentage fixation suppression (%FS) of caloric nystagmus were recorded by DC electrooculography (EOG), and stored directly onto FM magnetic tape. RESULTS: OKN velocities, pursuit gains, and %FS decreased toward the lesion side (group A), whereas OKN velocities and pursuit gains decreased toward the side contralateral to the lesion side. %FS decreased toward the lesion side (group B). MR images in group A showed that lesions were only in and near to the vestibular nuclei, and images in group B showed that lesions extended to the cerebellum. CONCLUSION: Visually guided eye movements allow classification of Wallenberg's syndrome into two types, one with brainstem lesions and one with both brainstem and cerebellar lesions. This demonstrates that assessing eye movements is helpful in supplementing MRI data. PMID- 12543155 TI - Changes in CAP adaptation in experimentally induced endolymphatic hydrops. AB - OBJECTIVE: To determine the effect of experimentally induced endolymphatic hydrops (ELHs) on compound action potential (CAP) adaptation. METHODS: The CAP was recorded 1 month after surgical treatment to produce ELH. The CAP threshold in response to click stimuli showed no significant differences between the hydropic and control groups. The CAP was elicited from the retroauricular-exposed round window using a silver ball electrode. Click trains consisting of 10 clicks with an interclick interval (Deltat) of either 4 or 8 ms were delivered through a loudspeaker placed in front of the animals' ears. The patterns of adaptation to these click trains were evaluated as functions of Deltat and intensity level. RESULTS: An abnormal pattern of CAP adaptation was observed in hydropic ears. CONCLUSIONS: These findings suggest that early ELH influences CAP adaptation. PMID- 12543156 TI - Zinc deficiency and tinnitus. AB - OBJECTIVE: To determine if there is a correlation between serum zinc levels and audiometric performance in tinnitus patients. METHODS: Seventy-three patients participated in this study. Patient's age was restricted to 20-59 years. All patients were examined at the otolaryngology outpatient clinic of the St. Marianna University Toyoko Hospital. The control group consisted of 38 age- and sex-matched healthy volunteers. A blood sample was taken to measure serum zinc levels. Hypozincemia was set at a level of the mean minus one S.D. in the control group. An average hearing sensitivity was calculated as the mean value of hearing thresholds at five frequencies: 250, 500, 1000, 2000, and 4000 Hz. Normal hearing was indicated when the hearing threshold at each of these frequencies was within 20 dB of normal thresholds. RESULTS: There was no significant difference in serum zinc levels between patients with tinnitus and controls. However, patients with tinnitus who had normal hearing had significantly lower serum zinc levels compared to controls. In contrast, no significant difference in serum zinc levels was found between patients with tinnitus who had hearing loss, and controls. A significant correlation between average hearing sensitivity and serum zinc level was observed. CONCLUSIONS: These findings suggest that zinc is involved in the generation of tinnitus, especially in patients whose hearing is relatively normal. PMID- 12543157 TI - Flexibility of vestibulo-ocular reflex adaptation to modified visual input in human. AB - The vestibulo-ocular reflex (VOR) serves to keep images relatively stable on the retina. To maintain appropriate performance and minimize image slip throughout life, VOR is subjected to long-term adaptive regulation by visual input. It has been reported that adaptive changes in VOR gain (eye velocity/head velocity) are evoked either by fitting subjects with magnifying, miniaturizing, or reversing spectacles during normal behavior, or by moving a large visual field in or out of phase relative to the subject's head movement. The changes in VOR gain are frequency selective. Here, we examine the extent of VOR gain flexibility by causing VORs of similar direction to undergo different behavioral gain changes. Nine healthy adults participated in the study, ranging in age from 24 to 38 years (mean: 26 years) and with no history of neurotological symptoms. All subjects were clinically normal according to a screening battery that included combined neurologic and otologic physical examinations. Horizontal and vertical eye positions were recorded by bitemporal DC-coupled electro-oculography (EOG). The subject sat in a rotating chair. The axis of rotation of the body was always earth-vertical, with the interaural axis crossing the axis of rotation of the chair. The head was pointed 20 degrees downwards in all experiments and stabilized in this position using a chin rest. The chair was surrounded by a half cylindrical optokinetic screen (78 cm in diameter) placed in front of the subject, onto which random dot patterns were projected. Goggles were used to ensure that the subject was in complete darkness during both pre- and postadaptation periods. The chair was rotated sinusoidally at maximum amplitude of 30 degrees or 60 degrees : for 30 degrees the stimulation was at 0.1, 0.2, 0.3, and 0.4 Hz; for 60 degrees it was at 0.1, 0.2, and 0.3 Hz. VOR adaptation was obtained by inducing a retinal slip velocity by short-term alteration of the visual input of the large field; this change was produced by a combination of sinusoidal head rotation and the random dot patterns. In each adaptation session, the sinusoidal head rotation was either at 0.1 or 0.3 Hz and the amplitude was 30 degrees. The random dot pattern was synchronized with sinusoidal head rotation in the same direction (x0 experiment) to make the retinal slip zero, and in the opposite direction to make the retinal slip twofold (x2 experiment). Therefore, a total of four adaptation protocols were tested. The subjects were asked to fixate on a single dot with the eyes straight-ahead in the x0 experiment, and follow it within random dot patterns in the x2 experiment. Each adaptation session lasted for 30 min. Two adaptation experiments were performed on each subject once per day. The averages of VOR gain and phase lag were calculated using Fourier analysis. Seven of the subjects who participated in the x2 adaptation experiment at 0.3 Hz and 30 degrees amplitude showed a steady increase of VOR gain within a couple of trials. One of the remaining two subjects showed a decrease in VOR gain in all the three trials, and the other subject showed a VOR gain increase in three trials and decrease in two trials. In the x2 adaptation experiment at 0.3 Hz and 30 degrees amplitude (peak velocity: 28 degrees /s), the percentage gain change ((post-pre)/pre) was 133% at the same stimulation and 100% at 0.4 Hz (peak velocity: 37 degrees /s). The gain change was 65% at 0.1 Hz and 60 degrees amplitude (peak velocity: 18 degrees /s), and 64% at 0.2 Hz and 60 degrees amplitude (peak velocity: 37 degrees /s). In the x1 adaptation experiment (30 degrees at 0.3 Hz), the percentage gain change was -62% under the same conditions, -50% at 0.1 Hz and 60 degrees amplitude, and -30% at 0.2 Hz and 60 degrees amplitude. No change of VOR gain was observed at other frequencies, and the subject was not adapted at 0.1 Hz and 30 degrees amplitude. The larger VOR gain increase in the x2 adaptation experiment for the faster head angular acceleration leads to the conclusion that the VOR gain mechanism can set vuite similar in terms of the head acceleration. PMID- 12543158 TI - Retrograde-labeling of pretecto-vestibular pathways in cats. AB - The projections of the nucleus of the optic tract (NOT) were studied in cats using three kinds of retrograde tracers (horseradish peroxidase wheat germ agglutinin (WGA-HRP), dextran fluorescein (DF) and dextran tetramethylrhodamine (DTR)). First, in the cases that WGA-HRP were injected into the rostral part of medial vestibular nucleus (VN), a significant number of retrograde-labeled neurons, which concentrated between 600 and 800 micrometer from the rostral part of the NOT, were observed in the NOT ipsilateral to the injection sites. Second, no double-labeled neurons were found following injections of DF in the medial VN and DTR in the nucleus prepositus hypoglossi (NPH) at the same time. In this experiment, it was made clear that two different kinds of neurons in the NOT projected to the medial VN and the NPH, respectively. PMID- 12543159 TI - Saccadic adaptation in the horizontal and vertical directions in normal subjects. AB - We studied the properties of adaptive gain control in the saccadic system and examined whether the adaptations in the horizontal direction transferred to those in the vertical direction, and vice versa, in 16 normal subjects. Eye movements were measured using search coil system. In the adaptive session, a target moved randomly in amplitude steps of 20 degrees or 30 degrees, with half of the subjects performing in the horizontal direction and the others in the vertical direction. The target eccentricity was changed by a constant percentage (70%) during each saccade aimed at the target. Gains were gradually decreased during the course of 120 repetitions, and then approached approximately 70%. Comparison was made of the accuracy (saccadic amplitude/first target step amplitude) of horizontal and vertical vectors of oblique saccades before and after the adaptation. After the horizontal adaptation, the accuracy of horizontal vectors decreased in the horizontal direction (P<0.05) but not in the vertical direction. After the vertical adaptation, however, the accuracy of horizontal and vertical vectors did not decrease. Our data show that adaptive gain control in the horizontal direction might not transfer to that in the vertical direction, nor vice versa. PMID- 12543160 TI - Horizontal eye response to an abrupt lateral head drop in normal subjects and patients with Meniere's disease. AB - OBJECTIVE: To detect the endolymphatic hydrops by using the otolithic nature in patients with Meniere's disease and delayed hydrops. METHODS: We developed a new technique for measuring horizontal eye movements, corresponding to lateral head drop in total darkness, approximately 1g environment. Normal subjects (n=16, mean age: 27.3 years old) and patients (seven for pre-operative state, four for furosemide test at pre-operative state; two for post-operative state) were employed in the test. The principle of the test is to drop the head in a lateral position separately from the body with a rapid release exposing the head. Bitemporal EOG, accelometric and trigger signals for the releaser were recorded for processing at 1 KHz sampling time. RESULTS: The drop was performed five times in succession. The latency of horizontal eye response was of 40 ms order in normal subjects on both sides and were never beyond 60 ms. Eye response of the patients was in different patterns from normal subjects and the latencies delayed either to normal side or to the affected side. Following furosemide intravenous injection, more certain information was obtained. Three of four patients were recovered to normal range as to the latency. CONCLUSIONS: We emphasize that this test may provide a new measure sensitive to the hydrops and can underline this test to detect the hydrops in Meniere's disease. PMID- 12543161 TI - Physical therapy for benign paroxysmal positional vertigo patients with movement disability. AB - OBJECTIVE: Particle repositioning manoeuvres are broadly recognized effective for benign paroxysmal positional vertigo (BPPV), however, we think that these therapies are not suitable for all BPPV patients. In this study we investigate whether these manoeuvres can be used for BPPV patients with the movement disability. In addition, we introduce our habituation training (HT) and report the results of treatment with it. METHODS: Nine BPPV patients with the movement disability due to orthopedic disease were examined. For each patient we determined their movement disabilities, and assessed the suitability of particle repositioning manoeuvres, Brandt-Daroff (B-D) manoeuvre and HT for them. RESULTS: All of these nine patients had neck or trunk movement disabilities due to orthopedic disease, and we judged that particle repositioning manoeuvres and B-D manoeuvre were impossible or too risky to be employed in them. Although HT was suitable for them and symptoms and signs of them improved within a short period of time. CONCLUSION: Particle repositioning manoeuvres were not suitable for BPPV patients with the movement disability because these manoeuvres required various movements. These patients were able to perform HT depending on their capabilities. We think that HT is caused by the fatigability of BPPV, and it accelerates the natural curing process. PMID- 12543162 TI - Patency of inferior meatal antrostomy. AB - OBJECTIVE: To evaluate the patency and effectiveness of inferior meatal antrostomy for the treatment of maxillary lesions. METHODS: Patients suffering from moderate-to-severe chronic sinusitis who underwent inferior meatal antrostomies at St. Marianna University Toyoko Hospital and followed up for at least 1 year after surgery were selected. Thirty sides of 27 patients were analyzed. Postoperative mucociliary function was examined in seven sides of seven patients. RESULTS: No closure was observed in all 30 procedures. Out of 30 antra, 26 (86.7%) had normal antral mucosa. Recurrent mucosal lesions inside the maxillary sinus were successfully treated through the inferior meatal window. No cases showed circular flow on the india ink test. All sinuses exhibited a discharge from the middle meatal window and three out of seven sinuses exhibited a discharge from the inferior meatal window. CONCLUSIONS: Although long-term follow-up is required to determine the patency of inferior meatal antrostomy, initial results are encouraging. The inferior meatal window provides both intra- and postoperative benefits. PMID- 12543163 TI - Effect of fexofenadine hydrochloride on cedar pollinosis. AB - OBJECTIVE: To investigate the therapeutic efficacy of fexofenadine hydrochloride (Allegra(R) tablets), an antihistaminic launched in 2001, in patients with cedar pollinosis by dividing them into two groups for comparison, i.e. the early treatment group in which treatment was started before the initial day of the pollen scattering, and the therapeutic-treatment group in which treatment was started after the initial day of the pollen scattering. METHODS: Early-treatment group: patients who visited the hospital before the initial day of cedar pollen scattering were orally given one tablet of the drug twice daily. Therapeutic treatment group: patients who visited the hospital after the initial day of cedar pollen scattering were orally given one tablet of the drug twice daily. The total number of cases in which the efficacy evaluation was possible was 37 cases (19 cases of the early-treatment group and 18 cases of the therapeutic-treatment group) after application of exclusion criteria. RESULTS: The useful rate of moderately effective or better against sneeze was 90% in the early-treatment group, and 78% in the therapeutic-treatment group, and there was a significant difference between both groups. The degree of satisfaction in the early-treatment group was 3.8 points, and 4.2 points in the therapeutic-treatment group, and the therapeutic-treatment group showed a higher score, but there was no significant difference between both groups. As adverse reaction, there was only one case of mild dizziness (2.7%), and no other adverse reactions such as sleepiness were observed. CONCLUSIONS: It was suggested that fexofenadine hydrochloride administered in patients with cedar pollinosis from before substantial pollen scattering might control their symptoms to mild ones, and might control worsening of their symptoms after the substantial pollen scattering, and, therefore, the drug was considered to be useful in early therapy. PMID- 12543164 TI - End-to-end anastomosis in chronic tracheal stenosis. AB - OBJECTIVE: This study assessed the efficacy and safety of end-to-end anastomosis of the trachea following segmental resection in chronic tracheal stenosis. METHODS: End-to-end anastomoses of the trachea were performed in 35 patients with chronic tracheal stenosis; 18 patients with tracheal invasion of thyroid cancer and 17 patients with long-term intubation and blunt injuries of the trachea. RESULTS: All operations were successful, except one whose unilateral recurrent nerve had not been identified in the recurrent thyroid cancer invasion with trachea. CONCLUSION: This operation provides a one-step cure for the stenosed trachea and can be applied to the resection of less than six tracheal segments. PMID- 12543165 TI - Video-assisted submandibular sialadenectomy using an ultrasonic scalpel. AB - OBJECTIVE: To evaluate the benefits in video-assisted submandibular sialadenectomy by using an ultrasonically activated scalpel (HS: Harmonic Scalpel(R), Ethicon, Somerville, NJ) and a special handmade lifting device. METHODS: The study cohort consisted of four male patients. We considered that the intraglandular part sialolithiasis of the submandibular glands was the main indicator for this technique. Under general anesthesia, a 15-20 mm skin incision was performed just above the inferior margin of the submandibular gland lying parallel to the mandible. We used a 4 mm diameter endoscope that we normally use for paranasal sinus surgery. Appropriate working space was created using a special handmade lifting device that was constructed from a wooden tongue depressor. By using HS, we dissected between the capsule and the circumferential tissue in order to minimize the possibility of damage to the facial nerve and lingual nerve. The submandibular gland was pulled out through the surgical wound, and Wharton's duct was then suture-ligated and resected. RESULTS: Endoscopic operation without recourse to conventional open surgery was achieved in all patients. No patients suffered any complications, such as functional disorder of the marginal mandibular, hypoglossal and lingual nerves, or wound hematoma formation. Excellent cosmesis was obtained in all patients. The amount of intraoperative bleeding ranged from 10 to 60 ml (mean: 38.0 ml). The operating time ranged from 175 to 250 min (mean: 232.5 min). CONCLUSIONS: We applied two useful devices to achieve the endoscopic submandibular surgery: an HS and a special handmade lifting device. Excellent cosmesis was obtained in all patients without complications. The disadvantage of this method is that the surgery is more time-consuming than conventional open methods. PMID- 12543166 TI - Sensitization of head and neck squamous cell carcinoma cells to Fas-mediated apoptosis by the inhibition of Bcl-X(L) expression. AB - BACKGROUND: Various types of malignant tumor cells are known to acquire resistance to Fas receptor (Fas)-mediated apoptosis. In Fas-sensitive cells, Fas mediated apoptosis is observed when anti-Fas antibody is bound to Fas. Bcl-2 and Bcl-X(L) are representative anti-apoptosis proteins reported to be capable of suppressing Fas-mediated apoptosis. OBJECTIVE: To investigate the mechanism of resistance acquisition to Fas-mediated apoptosis in cultured human head and neck squamous cell carcinoma cells (HNSCCs). METHODS AND RESULTS: We applied an anti Fas antibody (CH11) to Fas-expressing HNSCCs (HSC-2) and the CH11 did not induce cell death in HSC-2. Treatment with actinomycin D (ActD) converted the phenotypes of HSC-2 from CH11-resistant to CH11-sensitive. Western blot analysis showed no differences between ActD-treated and ActD-untreated HSC-2 in the expression of Bcl-2. On the other hand, the expression of Bcl-X(L) was greatly reduced in ActD treated HSC-2. Moreover, the reduction of Bcl-X(L) by specific antisense oligonucleotide treatment enhanced the CH11-induced cell death of HSC-2. CONCLUSION: Our data suggest that Fas-signaling might be regulated by a Bcl-X(L) inhibitable step in CH11-resistant HSC-2. PMID- 12543167 TI - Inhibition of caspase-9 activity and Apaf-1 expression in cisplatin-resistant head and neck squamous cell carcinoma cells. AB - We have previously reported that cisplatin induces caspase-9 (Casp9) activation in head and neck squamous cell carcinoma cells in vitro (HNSCCs). Our purpose here was to examine whether HNSCCs selected for resistance to cisplatin fail to exhibit Casp9 activation in response to cisplatin. The cisplatin-resistant HNSCCs (HSC-2CR) were selected from cisplatin-sensitive HNSCCs (HSC-2) for growth in the presence of cisplatin. Following cisplatin treatment, protelyzed Casp9 subunits were detected in HSC-2, but not detected in HSC-2CR. Using a direct enzymatic assay measuring cleavage of the synthetic peptide substrate (LEHD-AFC), Casp9 activity in cisplatin-treated HSC-2CR was less than that in cisplatin-treated HSC 2. Apoptotic protease-activating factor 1 (Apaf-1) has been shown to participate as an adaptor molecule in Casp9 activation. In the presence of cytochrome c (Cyt c) released from mitochondria, Apaf-1 binds to Casp9 and causes its activation. HSC-2 expressed 2-fold higher levels of Apaf-1 compared with HSC-2CR. On the other hand, following cisplatin treatment, the same degree of increase in cytoplasmic Cyt c was detected in both HSC-2 and HSC-2CR. These results suggest that in a certain type of HNSCCs, the inhibition of Casp9 activity and Apaf-1 expression may represent a mechanism of acquired cisplatin resistance. PMID- 12543168 TI - Vestibular-evoked myogenic potentials in two patients with Ramsay Hunt syndrome. AB - We report on the function of the inferior vestibular nerve, as monitored by the vestibular-evoked myogenic potentials (VEMP), in two patients suffering from Ramsay Hunt syndrome. Both the patients presented canal paresis (CP) and hearing loss, but in one patient normal VEMP was recorded while the other presented vagus nerve paralysis plus no VEMP response at the highest stimulus intensity used in our institute (i.e., 105 dB nHL). PMID- 12543169 TI - Neurotological findings in a patient with narrow internal auditory canal: a case report. AB - We report neurotological findings in a patient with unilateral narrow internal auditory canal, as confirmed by computed tomography. The patient presented no auditory brainstem response on the affected side. Vestibular tests including vestibular-evoked myogenic potentials (VEMP) and caloric test revealed normal function of both inferior and superior vestibular neural pathways. PMID- 12543170 TI - Pediatric bilateral facial nerve palsy secondary to acute otitis media. AB - Bilateral facial palsy after the onset of acute otitis media is presented in a 19 month-old patient. Facial nerve palsy disappeared gradually after myringotomy. The mechanisms underlying the facial nerve palsy with otitis media are discussed. PMID- 12543171 TI - A case of carcinoid tumor of the middle ear. AB - We report here a case of a carcinoid tumor observed in the middle ear (ME), which was initially diagnosed as ME adenoma. The patient was a 64-year-old woman who was first seen in our hospital in March 2001 for a 7-month hearing loss. On otoscopic examination, a whitish mass could be observed through the intact tympanic membrane. High-resolution computed tomography demonstrated a tumor-like lesion in the ME with no evidence of bone destruction. A myringotomy and biopsy were performed and an initial diagnosis of ME adenoma was made. Light microscopy showed fragments of cellular tissue in which both glandular (adenomatous) and trabecular (carcinoid) growth patterns could be identified, but neuroendocrine differentiation was not detected by immunohistochemistry (negative staining for chromogranin A and synaptophysin). On the basis of this diagnosis, the patient underwent a tympanomastoidectomy in June 2001, in which the presumed ME adenoma was completely excised and the diagnosis was modified to ME carcinoid tumor. Immunohistochemical examinations at that time showed positive staining of the tumor cells for chromogranin A and synaptophysin. This case suggests the difficulties in distinguishing ME carcinoid tumors from ME adenomas. The patient is without recurrence of her disease to date. PMID- 12543172 TI - A case of meningoencephalic herniation of the temporal bone. AB - A case of cerebrospinal fluid leakage from a meningoencephalic herniation of the temporal bone is presented. Computed tomography and magnetic resonance imaging were useful for a diagnosis of meningoencephalic herniation. After surgical treatment, no recurrence was observed during a 6-month follow-up period. PMID- 12543173 TI - Powered endoscopic marsupialization for recurrent sphenoid sinus mucocele: a case report. AB - We report a case of recurrent sphenoid mucocele successfully treated by using a powered instrument under endoscopic control. A 59-year-old male came to our clinic complaining of severe headache, right-side facial numbness (in the areas of the first and second branches of the trigeminal nerve), diplopia, and right blepharoptosis. Computed tomography (CT) imaging revealed opacification and expansion of the sphenoid sinus lesion. The lesion was diagnosed as right-side sphenoid mucocele affecting the functions of the trigeminal (first and second branches), oculomotor, and abducent nerves. Endoscopic drainage of the right-side sphenoid mucocele leads to gradual improvement of these symptoms. Approximately 1 year after the drainage procedure, the size of the enlarged sphenoid sinus ostium had decreased. The patient underwent endoscopic right-side total marsupialization of the sphenoid sinus using a powered instrument. Subsequently, the patient has presented no evidence of recurrent disease after 1 year of follow-up. PMID- 12543174 TI - A rare case of olfactory neuroblastoma with intracranial extension. AB - Olfactory neuroblastoma (ONB) is a rare form of head and neck tumor, and further ONB presenting an intracranial extension is extremely rare. Therefore, at the present time there is no established therapeutic standard supported by oncological studies. Our report documents a patient who underwent a craniotomy in combination with transnasal approach for tumor excision, followed by postoperative irradiation. We conclude that the patient shows no evidence of recurrence during the 11-year follow-up involving sinus CT scans and endoscopic examinations. PMID- 12543175 TI - Two cases of Aspergillus sinusitis with bone destruction. AB - We experienced the clinical courses of two cases of destructive Aspergillus in paranasal sinuses in which an Aspergillus fungus ball was formed, and which caused bony destruction are described. The case in which a nidus could not be completely surgically removed and with intervening diabetes had an unfavorable prognosis. It is recommend to perform an early expanded operation as well as to administer a sufficient amount of antifungal drugs for the treatment of destructive Aspergillus in paranasal sinuses. PMID- 12543176 TI - Cementifying fibroma in the ethmoidal sinus extending to the anterior cranial base in an 11-year-old girl: a case report. AB - A case of an 11-year-old female with cementifying fibroma in the ethmoidal sinus extending to the anterior cranial base was presented. She complained of slowly progressive nasal obstruction and anosmia. Magnetic resonance imaging (MRI) revealed a tumor in both ethmoidal and sphenoidal sinuses and nasal cavity. Although a tumor reduction surgery was attempted through a transnasal approach, tumor size increased gradually and right exophthalmos subsequently appeared. The pathological diagnosis was cementifying fibroma of the nose. Total tumor removal was consequently accomplished through the intracranial anterior transbasal approach. The surgical approach should be selected according to the site and expansion of the lesion in individual patients. PMID- 12543177 TI - A case of an inverted tooth in the nasal cavity. AB - We report a case of an inverted tooth in the nasal cavity. The patient was a 27 year-old man who attended our hospital in May 1998, complaining of left cheek pain. There was nothing remarkable in his medical or family history. Fiberscopic (intranasal) and radiological examinations revealed a white foreign body in the left nasal cavity, within 2 cm of the left nostril. This foreign body was diagnosed as an inverted tooth. It was removed under general anesthesia and found to be 17 mm in length. Although the tooth showed a single root, it possessed two cusps and we deduced it to be a molar. PMID- 12543178 TI - A case report of difficult diagnosis in the patient with advanced laryngeal tuberculosis. AB - We report a patient with severe laryngeal tuberculosis (LTB) involving thyroid cartilage and combined with whole-bone metastasis. A 57-year-old male had presented only with hoarseness. Radiological findings were indicative of suspected metastasis from a malignant tumor. However, tuberculosis was considered by histopathological findings, and so sputum samples were tested for acid-fast bacilli and purified protein derivatives of tuberculin in order to detect the presence of LTB. A polymerase chain reaction confirmed the diagnosis. Anti tuberculous medications were effective in resolving the hoarseness, and the removal of the mass in the right wing of thyroid cartilage was confirmed by computed tomography (CT). PMID- 12543179 TI - Subcutaneous emphysema secondary to tonsillectomy: a case report. AB - We report a patient in whom subcutaneous emphysema developed shortly after a tonsillectomy. A 55-year-old female with a chronic tonsillitis underwent a tonsillectomy, and about 8 h after surgery complained of swelling to the left side of the face and neck. There was crepitus and local tenderness in the left side of the neck. A CT scan revealed subcutaneous emphysema. The mechanisms underlying emphysema after tonsillectomy are discussed. PMID- 12543180 TI - A case of large dermoid cyst in the floor of the mouth. AB - Dermoid cysts in the floor of the mouth are rarely observed. When they develop, they do not appear until they grow large enough or appear through infection. Some operative approaches and management have been performed to large dermoid cysts in the floor of the mouth with some variations. Intra-oral approach is the most useful for cosmetic appearance. We presented a 24-year-old female who underwent expedient surgical excision using finger end serviceable enucleation through the intra-oral approach. PMID- 12543181 TI - A case of relapsing polychondritis involving placement of an expandable metallic stent. AB - Relapsing polychondritis is a relatively uncommon disease characterized by recurrence of progressive inflammation of cartilaginous structures. Laryngotracheal involvement occurs in 50-70% of cases, and occasionally results in acute airway obstruction. We reported a case of relapsing polychondritis with severe tracheal stenosis that was treated by the placement of two expandable metallic stents. A 58-year-old man was hospitalized with fever and breathing difficulties. A tracheostomy was performed and a silicone T-tube was placed. A diagnosis of relapsing polychondritis was made based on the biopsy from auricular and tracheal cartilages. Two expandable metallic stents were placed in the trachea to bilateral main bronchus. This dilated the airway lumen and resulted in the dyspnea disappearing completely. However, 18 months later a sudden massive hemorrhage occurred through the tracheostoma, and he died of respiratory failure. Autopsy showed a tracheoinnominate artery fistula. We discuss the management of relapsing polychondritis with severe tracheal stenosis. PMID- 12543182 TI - Penetrating neck injuries involving wooden foreign bodies: the role of MRI and the misinterpretation of CT images. AB - A 12-year-old, boy fell in the bush and suffered a penetrating neck wound. Computed tomography(CT-scan) was performed, but no foreign body was detected inside the wound. The patient returned home after ordinary wound care. About 6 months after the accident, the patient visited hospital, because of inflammatory neck pain. A CT-scan was performed and the foreign body appeared as a high density. Magnetic resonance imaging (MRI) revealed the presence of a large foreign body. Under general anesthesia the wooden (bamboo) foreign body was removed. This case highlights the needs to be aware of the potential for misinterpretation of CT images, especially in acute wooden penetration injury. Its also demonstrates that MRI is a useful adjust to CT in the detection of non metallic foreign bodies. PMID- 12543183 TI - Rare localization of paraganglioma in head and neck. AB - In this paper, we describe the clinical course of a 61-year-old female patient with paraganglioma in the head and neck region. Computed tomographic scan (CT), magnetic resonance imaging (MRI), ultrasound scan (US) and arteriogram findings initially led us to suspect that this tumor originated in the vagal nerve. In particular, a color Doppler US enabled an easy diagnosis of hypervascular tumor. We removed this surgically, but the tumor was easy to peel from the vagal nerve and carotid bifurcation. The distal side of the tumor was under the digastric muscle and running into the hypoglossal nerve. The intraoperative findings were highly suggestive of localization at the hypoglossal nerve, although Xth and XIIth cranial nerve palsies have remained. PMID- 12543184 TI - New color vision tests to evaluate faulty color recognition. AB - PURPOSE: To develop and assess new color vision tests to be used in evaluating faulty color recognition. METHODS: We developed new color vision tests to evaluate faulty color recognition. The two types of color vision tests, designed to assess faulty color recognition in color vision deficiencies, are based on principles that are different from those of the conventional color vision tests. In the first test plate, the subject is asked to choose either a red, green, or gray line from among 10 lines that are randomly colored red, green, gray, yellow, or blue. The score is the difference between the number of correct answers and the number of incorrect answers. In the second test plate, the subject is asked to identify a total of 10 red azalea blossoms, which are dispersed among numerous green leaves. Seventy-five persons with congenital color deficiencies and 20 subjects with normal color vision were examined using these new test plates. RESULTS: The scores differed significantly between dichromats and anomalous trichromats, and between anomalous trichromats and subjects with normal color vision. CONCLUSIONS: The new tests are easy to use, sensitive, and have good reproducibility for use in discriminating subjects with color vision anomalies. These tests reveal the faulty color recognition that occurs unconsciously in persons with color deficiencies, and are useful in judging the quantification of color vision required in their daily life and occupations. PMID- 12543185 TI - Identification and characterization of beta-D-galactosyl-transferase in chick corneas. AB - PURPOSE: To purify beta-galactosyltransferase, which is involved in the biosynthesis of keratan sulfate, from 2-day-old chick corneas. METHODS: The activity was assayed using pyridylaminated GlcNAcbeta1-3Galbeta1-4Glc as acceptor substrate. RESULTS: The beta-galactosyltransferase did not bind to several ion exchange and affinity columns. In particular, it did not bind completely to an alpha-lactalbumin-agarose column. The partially purified enzyme showed an optimum pH at 7.0 [(N-[2-hydroxyethyl]piperazine-N'-[2-ethanesulfonic acid]) (HEPES) buffer]. Pyridylaminated GlcNAcbeta1-3Galbeta1-4GlcNAc, as well as the pyridylaminated GlcNAcbeta1-3Galbeta1-4Glc, served as the acceptor substrate of the enzyme, but not p-nitrophenyl-beta-GlcNAc. The crude extract from chick corneas contained a high activity of the beta-galactosyltransferase, which transfers Gal to pyridylaminated 6-sulfo-GlcNAcbeta1-3Gal, but the activity was almost all lost during the purification procedures. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of the enzyme fraction showed many bands. When each of the five main band proteins was partially analyzed for the amino acid sequence, none showed homology with the recently reported chick beta galactosyltransferases 1 and 2. CONCLUSIONS: beta-Galactosyltransferase, which transfers Gal to 6-sulfo-GlcNAc end, was identified in chick corneas. The chick corneal beta-galactosyltransferase(s) may be a novel one. PMID- 12543186 TI - Endogenous substance P in corneal epithelial cells and keratocytes. AB - PURPOSE: To detect endogenous substance P (SP) and neurokinin receptor 1 (NK1R) in cultured human corneal epithelial cells (HE) and cultured human keratocytes (HK). METHODS: Messenger RNA (mRNA) expression of SP and endogenous SP in HE and HK was investigated by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay. mRNA expression of NK1R in HE and HK was investigated by RT-PCR. RESULTS: The mRNA expression of SP and endogenous SP were recognized in HE and HK. Furthermore, the mRNA of NK1R was expressed in HE and HK. CONCLUSION: It was suggested that endogenous SP regulates the biological functions of HE and HK in autocrine or paracrine fashion. PMID- 12543187 TI - Neuroprotective effects of R(-)-1-(benzo[b]thiophen-5-yl)-2- [2-(N,N diethylamino)ethoxy]ethanol hydrochloride (T-588) against retinal ganglion cell death induced by elevated intraocular pressure in rat. AB - PURPOSE: We investigated whether T-588 can attenuate retinal ganglion cell (RGC) death induced by elevated intraocular pressure (IOP). METHODS: IOP elevation was induced unilaterally by argon laser irradiation of the rat trabecular meshwork 4 days after an intracameral injection of India ink. We orally administered either the vehicle, or 10, 30, or 100 mg/kg body weight (BW) of T-588 24 hours before the laser application. Five days after the laser application, 1.5 microL of 3% Fast Blue was injected into the superior colliculi bilaterally. Three days after the Fast Blue injection, the eye was enucleated and the retinal whole flatmount was prepared. Labeled ganglion cells were counted by fluorescence microscope with an ultraviolet filter. RESULTS: Laser treatment significantly increased the IOP. The percentages of labeled RGCs in the lasered eyes as compared with the nonlasered contralateral eyes were 78.0 +/- 11.6% in the control group, 78.7 +/- 12.9% in the 10 mg/kg BW group, 79.1 +/- 13.0% in the 30 mg/kg BW group, and 91.0 +/- 9.0% in the 100 mg/kg BW T-588-treated group. The survival rate of RGCs was significantly higher in the 100 mg/kg BW T-588-treated group than in the control group. CONCLUSION: T-588 appears to have a neuroprotective effect on retinal ganglion cells in this ocular hypertensive model. PMID- 12543188 TI - An automated measuring system for fundus perimetry. AB - PURPOSE: In order to automate fundus perimetry, which is usually laborious because of the many manual operations and the frequent eye movements, we developed a new fundus perimeter with an automated measuring system using computer technology. METHODS: This fundus perimeter consists of an infrared fundus camera, a fundus-pursuit unit, and a target-display unit. Both units are personal computers operating on Windows. After presetting the measuring points on the fundus, perimetry is automatically performed. The fundus-pursuit unit recognizes and pursues the fundus reference region while calculating its coordinates in real time during perimetry. The target-display unit corrects the stimulus coordinates with data from the fundus-pursuit unit and even if eye movement occurs, automatically displays the stimulus targets at the correct retinal point that had been preset. RESULTS: Fundus perimetry can be automatically and precisely performed with this automated measuring system. It shortens the duration of the examination and frees the operator from the lengthy and laborious manual operations during perimetry. CONCLUSIONS: This automated measuring system will make fundus perimetry easier for the operator, as well as more popular for clinical use. PMID- 12543189 TI - Mitogen-activated protein kinase inhibitor, PD98059, inhibits rat retinal pigment epithelial cell replication by cell cycle arrest. AB - PURPOSE: To investigate the effect of PD98059, a mitogen-activated protein kinase (MAPK) inhibitor, on the replication of rat cultured retinal pigment epithelial (RPE) cells. METHODS: Growth-phase rat RPE cells were exposed to various concentrations of PD98059 in serum-free F12 medium containing 0.1% dimethyl sulfoxide. Cell proliferation was assessed by cell counts using a hemocytometer. Cell viability was tested by CellTiter 96 AQueous Non-Radioactive Cell Proliferation Assay at 24 hours after PD98059 application. Hoechst 33552 and propidium iodide staining were used to assess nuclear morphology. Immunostaining with Ki67 antibody was used for cell cycle analysis because the staining patterns produced on cells are characteristic depending on their position within the cell cycle. RESULTS: PD98059 inhibited cellular proliferation of cultured rat RPE cells in a dose-dependent manner but did not induce cell death. Twenty-four hours after the application of PD98059, cultured RPE cells were not immunopositive for Ki67, indicating that their cell cycle was arrested in the G0/G1 phase. CONCLUSION: These results demonstrated that MAPK inhibition arrested cell cycle progression of rat cultured RPE cells at the G0/G1 phase. The pharmacological induction of cell cycle arrest could be a new approach to inhibit cellular proliferation in such conditions as proliferative vitreoretinopathy. PMID- 12543190 TI - Colvard pupillometer measurement of scotopic pupil diameter in emmetropes and myopes. AB - PURPOSE: To prospectively compare the scotopic pupil size between emmetropes and myopes using a Colvard pupillometer. METHODS: The pupil diameters of 55 normal subjects and 55 healthy myopic subjects were measured with the Colvard pupillometer in a low-light situation that simulated the level of light encountered while driving at night. RESULTS: The mean (+/- SD) age of the emmetropic subjects was 30.78 years +/- 10.03 (range, 18-54 years) and the mean (+/- SD) age of the myopic subjects was 27.35 years +/- 8.43 (range, 21-52 years). The mean (+/- SD) scotopic pupil diameter was 6.46 +/- 0.90 mm (range, 4.5-8.0 mm) in the emmetropic group and 6.98 +/- 0.67 mm (5.5-8.5 mm) in the myopic group. The unpaired Student t-test showed that the difference in the scotopic pupil diameter between emmetropes and myopes was statistically significant (P =.0001). CONCLUSIONS: The mean scotopic pupil diameter in myopes was larger than that in emmetropes. Therefore, a large ablation zone of the cornea or an appropriate optical size of the phakic intraocular lens should be considered in refractive surgery. Preoperative scotopic pupil measurements may be necessary in all refractive patients. PMID- 12543191 TI - Underestimation of intraocular pressure in eyes after laser in situ keratomileusis. AB - PURPOSE: Retrospectively, we reviewed the records of 65 patients (115 eyes) regarding the intraocular pressure (IOP) after laser in situ keratomileusis (LASIK). METHODS: The mean patient age was 31.2 +/- 10.5 years. The average preoperative spherical equivalent was -6.85 +/- 2.54 diopters. A noncontact pneumatic tonometer and a Goldmann applanation tonometer were used in measuring the IOP. RESULTS: The IOP after LASIK was significantly lower than that before surgery (Mann-Whitney U-test). The IOP correlated significantly with the corrected diopter value, corneal thickness, and corneal curvature (Spearman rank correlation). CONCLUSIONS: The postoperative IOP can be underestimated due to decreased corneal thickness and curvature. When evaluating IOP after LASIK surgery, this possibility should be carefully investigated. PMID- 12543193 TI - A case of cystoid macular edema associated with latanoprost ophthalmic solution. AB - BACKGROUND: Although there have been reports of adverse effects after use, it is unclear whether latanoprost ophthalmic solution contributes to the development of cystoid macular edema (CME). CASE: A 71-year-old man underwent lens extraction, the insertion of an intraocular lens, and vitrectomy for elevated intraocular pressure (IOP) associated with lens subluxation in the left eye. After the surgery, antiglaucoma ophthalmic solutions controlled IOP well for over a year, maintaining good visual acuity with no abnormalities in the fundus. OBSERVATIONS: Two months after the previously prescribed antiglaucoma ophthalmic solutions were replaced by latanoprost, the patient's visual acuity decreased and CME developed. When latanoprost was replaced by other antiglaucoma ophthalmic solutions for controlling IOP, CME disappeared and visual acuity returned to the base level. CONCLUSIONS: Latanoprost may be involved in the development of CME. Patients who have undergone vitreous surgery or those with aphakia should be carefully observed for the possible development of CME associated with the use of latanoprost, even a long time after surgery. PMID- 12543192 TI - Use of scanning laser ophthalmoscope microperimetry in clinically significant macular edema in type 2 diabetes mellitus. AB - PURPOSE: We used scanning laser ophthalmoscope (SLO) microperimetry to evaluate scotomas in patients with clinically significant diabetic macular edema (CSME) in type 2 diabetes mellitus. METHODS: We studied 19 patients (mean age = 63 years; range, 45-78 years) (19 eyes). SLO microperimetry was performed in all eyes. We divided patients into three groups as follows: dense scotoma, relative scotoma, and no scotoma. The following variables were documented: age; duration of diabetes, hemoglobin A(1c) levels; logarithm of the minimum angle of resolution (Log(MAR)) visual acuity; refractive power; a history of panretinal photocoagulation; presence or absence of proliferative diabetic retinopathy, vitreomacular separation, and cystoid changes; the type of macular edema; and stability of fixation. All variables were compared in the three groups. RESULTS: We identified 4 eyes (21.1%) with dense scotoma, 10 (52.6%) with relative scotoma, and 5 (26.3%) with no scotoma. There were significant differences in log(MAR) visual acuity among those with dense scotoma (1.4 +/- 0.5), relative scotoma (0.6 +/- 0.2), and no scotoma (0.2 +/- 0.3) (P <.05), and in the prevalence of cystoid changes, diffuse edema, and unstable fixation among those with dense scotoma (75%, 75%, and 100%, respectively), relative scotoma (20%, 30% and 50%, respectively) and no scotoma (0%, 0% and 0%, respectively) (P <.05). CONCLUSIONS: Macular scotoma was observed by SLO microperimetry in 74% of the patients in this study. A scotoma in CSME is related to the formation of cystoid changes and the type of macular edema. In eyes with CSME in type 2 diabetes mellitus, a scotoma in the macula causes visual acuity impairment and unstable fixation. PMID- 12543194 TI - Macular nerve fibers temporal to fovea may have a greater potential to recover function in patients with Leber's hereditary optic neuropathy. AB - PURPOSE: It is known that even after visual loss, younger patients with Leber's hereditary optic neuropathy (LHON) can recover vision. The purpose of this study was to determine the mean age at onset for LHON patients with and without visual recovery who carried the 11778 mutation, and to determine the pattern of central vision recovery. METHODS: Thirty-five LHON patients with the 11778 mutation of mitochondrial DNA who had visited the Keio University Hospital between 1980 and 1999 and were followed for 2 to 20 years, were the subjects of this retrospective study. The patients who had recovered vision were tested by Goldmann perimetry, Humphrey perimetry, and landmark-driven fundus microperimetry with a scanning laser ophthalmoscope (SLO). The fixation status was assessed by SLO microperimetry. RESULTS: Nine of the 35 patients (14 of 70 eyes) demonstrated a recovery of visual acuity to better than 0.3 in at least one eye. The mean age of disease onset was 15.9 +/- 4.6 years in patients with visual recovery and 25.5 +/ 8.9 years in patients without visual recovery. This difference in the mean age at onset was significant (P =.0001; Welch t-test). These 9 patients (14 eyes) showed fenestrated central scotomas in testing by Humphrey 10-2 threshold and SLO microperimetry. The nasal side of the central visual fields had a higher sensitivity than the temporal side in 7 of the 9 patients in Humphrey 10-2 threshold testing. Areas insensitive to 0 dB were detected on the nasal side of the central retina in these patients by SLO microperimetry, and fixation stability was related to the degree of clinical visual acuity. CONCLUSION: The LHON patients with the 11778 mutation and a younger age of onset were more likely to show visual recovery. The findings made by perimetry suggest that the nerve fiber bundles in the nasal field (retina temporal to the fovea) may have a greater potential to recover function in LHON patients. PMID- 12543195 TI - Pseudotumor cerebri induced by minocycline therapy for acne vulgaris. AB - BACKGROUND: We report a case of a young girl who developed pseudotumor cerebri while taking minocycline for acne vulgaris. CASE: A 16-year-old girl without a history of menstrual irregularity, weighing 60 kg (body mass index: 26.0%) presented with a history of 1 week of headache and sudden onset of a horizontal diplopia. OBSERVATION: Examination revealed bilateral papilledema and an abduction deficit in her right eye. Her cerebrospinal fluid had an opening pressure of 400 mm H(2)O and a normal composition. Following normal findings on computed tomography and magnetic resonance imaging, a diagnosis of pseudotumor cerebri was made. She reported receiving minocycline to treat acne vulgaris during the previous 3 weeks. The headache resolved with withdrawal of minocycline. The diplopia and papilledema resolved after two lumbar punctures, although the visual field defects persisted. The minocycline concentrations in the serum and cerebrospinal fluid taken after cessation of the drug were below the detectable level. CONCLUSION: The role of minocycline should be considered and routine ophthalmologic examination during minocycline treatment should be performed when pseudotumor cerebri occurs in patients treated for acne vulgaris. PMID- 12543196 TI - Heterozygous Ala137Pro mutation in keratin 12 gene found in Japanese with Meesmann's corneal dystrophy. AB - PURPOSE: To report the molecular genetic analysis of a Japanese pedigree with Meesmann's corneal dystrophy (MCD). METHODS: Sequencing of the keratin 3 and keratin 12 genes was performed in 2 patients who were siblings and in an unaffected individual in the same family. The patients had the typical corneal microcysts and recurrent erosions with mild photophobia. RESULTS: A novel mutation resulting in the substitution of alanine to proline in codon 137 of the keratin 12 gene (Ala137Pro) was found in the 2 patients, but not in the unaffected member of the family and the 50 controls. CONCLUSIONS: This novel mutation (Ala137Pro) of the keratin 12 gene found in a Japanese family had caused MCD. PMID- 12543197 TI - Immunohistochemical study of epiretinal proliferative cellular tissue from a patient with sarcoidosis. AB - BACKGROUND: Sarcoidosis is a granulomatous disease causing uveitis. Although steroid therapy is usually effective for many patients, some are resistant to this therapy. In such cases, vitrectomy may be chosen as the therapeutic method to treat intraocular inflammation. CASE: A 26-year-old Japanese man was diagnosed as having sarcoidosis by clinical findings and histological examination of bronchoalveolar lavage. OBSERVATION: One year after diagnosis, vitreous opacity worsened and pars plana vitrectomy (PPV) was performed. Six months later, recurrence of vitreous opacity appeared and severe retinal exudative changes with proliferative vitreoretinopathy developed. One year after the first operation, PPV was repeated and the epiretinal proliferative tissue was removed and examined. CONCLUSIONS: Histologic examination of the specimen showed scattered noncaseating granulomatous inflammation mainly consisting of epithelioid histiocytes and lymphocytes. Plasma cells as well as T cells were identified and the predominance of CD8-positive T cells was demonstrated. Taking everything into consideration, a new finding of CD8 predominance in the epiretinal proliferative tissue was demonstrated. PMID- 12543198 TI - Surgical treatment of limbal vernal keratoconjunctivitis by resection of a limbal lesion. AB - PURPOSE: To report our experience managing a limbal mass associated with limbal vernal keratoconjunctivitis (VKC) by surgical excision. METHODS: A 16-year-old girl was referred to our hospital with the chief complaints of itching and photophobia in both eyes. She had a 2-year history of gradually enlarging limbal gelatinous masses on the temporal limbus in both eyes, which were diagnosed as limbal papillae of VKC. As the symptoms in her left eye were more severe, surgical resection of the limbal mass in her left eye was performed to relieve mechanical stress on the eyelid, followed by a free conjunctival autograft taken from the supranasal bulbar conjunctiva. RESULTS: After excision of the mass, ocular inflammation and other symptoms gradually disappeared. At 12-month follow up, there was no recurrence of the mass. Findings in a histological examination of the excised limbal lesion were consistent with those of limbal papilla of VKC. CONCLUSION: In recalcitrant cases of limbal VKC, in which the symptoms are apparently caused by an elevated limbal mass, surgical excision of the limbal mass can be one of the therapeutic modalities, and it may facilitate resolution of the symptoms caused by chronic limbal VKC. PMID- 12543199 TI - Frosted branch angiitis associated with Harada disease-like manifestations recurs 10 years later. AB - PURPOSE: To describe a case with frosted branch angiitis occurring in association with manifestations of Harada disease. CASE: A 41-year-old man developed marked iridocyclitis, frosted branch angiitis and retinal white dots, together with serous detachment of the sensory retina in the posterior pole of both eyes. OBSERVATIONS: Fluorescein angiography demonstrated multiple retinal pigment epithelial leakage points in the early phase followed by late dye pooling in the serous detachment, consistent with the manifestations of Harada disease. The patient experienced recurrence of the same combination of manifestations 10 years later. On both occasions, he showed mononuclear cell increase and protein elevation in the cerebrospinal fluid on spinal tap. CONCLUSION: Fluorescein angiography and spinal tap supported the diagnosis of Harada disease in the patient. The present patient could be diagnosed as having either Harada disease with frosted branch-like response or frosted branch angiitis with serous retinal detachment as occurs in Harada disease. PMID- 12543214 TI - Effects of tryptophan depletion on acute smoking abstinence symptoms and the acute smoking response. AB - Given the putative role of serotonin in the modulation of smoking withdrawal and the central actions of nicotine, this study examined the affective and neuroelectric correlates of smoking abstinence and cigarette smoking following depletion of the serotonin precursor, tryptophan. In a randomized, double-blind two session (tryptophan depletion [TD] vs. nondepletion), placebo-controlled design, spectrally analyzed electroencephalogram (EEG), self-ratings of withdrawal symptoms and mood states were assessed in 18 male cigarette smokers before smoking abstinence, 5 h postsmoking abstinence and again following sham smoking and the smoking of one cigarette. Compared to a nutritionally balanced amino acid (AA) mixture containing tryptophan (i.e., placebo mixture), oral ingestion of a similar mixture devoid of tryptophan resulted in a 70% reduction of plasma tryptophan but failed to alter the appearance or reversal (by acute cigarette smoking) of withdrawal symptoms, negative mood states and increased slow wave EEG in male smokers deprived of cigarettes. These results, although not supporting a role for the serotonergic system in acute smoking and early smoking abstinence symptoms, are discussed in relation to the neuropharmacology of smoking behavior and suggestions for future work. PMID- 12543215 TI - Inhibition of acute hyperammonemia-induced convulsions by systemically administered gamma aminobutyric acid in rats. AB - The present study has investigated the effects of intraperitoneally administered gamma aminobutyric acid (GABA) on ammonium chloride-induced hyperammonemia and convulsions in rats. Systemically administered GABA did not alter the concentration of GABA in the brain of control as well as hyperammonemic animals. However, hyperammonemia-induced convulsions were inhibited by GABA in a dose dependent manner. This was accompanied by a dose-dependent decrease in the concentrations of ammonia in both blood and brain and an elevation of glutamine in the blood. These results suggest that GABA has the potential to prevent acute hyperammonemia by increasing detoxification of ammonia to glutamine. As a result, the diffusion of ammonia from blood into the brain has been decreased. This accounts for an inhibition of convulsions by systemically administered GABA in hyperammonemic animals. PMID- 12543216 TI - Chronic access to a sucrose solution enhances the development of conditioned place preferences for fentanyl and amphetamine in male Long-Evans rats. AB - Consumption of palatable food and fluids alters the behavioral consequences of psychoactive drugs. To further investigate the effects of intake of palatable nutrients on the rewarding properties of these drugs, the effects of chronic intake of a sweet sucrose solution on the development of conditioned place preferences (CPP) to a mu-opioid agonist, fentanyl, and to a stimulant drug, amphetamine, were examined. Male Long-Evans rats consumed laboratory chow and water or chow, water, and a 32% sucrose solution. CPP testing was conducted in a three-chamber apparatus. In Experiment 1 (over four conditioning days), rats received saline, 0.004, or 0.016 mg/kg sc fentanyl citrate before being placed on the nonpreferred side of the apparatus and saline (subcutaneously) before being placed on the preferred side during a separate session on the same day. When given access to all three chambers, rats injected with 0.016 mg/kg fentanyl spent significantly more time on the drug-paired side than rats injected with saline. Furthermore, sucrose-fed rats displayed a significantly greater CPP than chow-fed rats. After conditioning, rats were tested for fentanyl-induced antinociception using the tail-flick test. Using a cumulative dose procedure, fentanyl (0.003, 0.010, 0.030, and 0.100 mg/kg sc) led to dose-dependent increases in tail-flick latencies. Rats fed with sucrose displayed significantly greater responses to fentanyl than those in the chow group. In Experiment 2, rats spent significantly more time on the drug-paired side of the CPP apparatus following injections of 0.33 or 1.0 mg/kg amphetamine than after saline injections. Additionally, following injection of 0.33 mg/kg amphetamine, sucrose-fed rats spent significantly more time on the drug-paired side of the chamber than chow-fed rats. PMID- 12543217 TI - Gender differences in the intravenous self-administration of mu opiate agonists. AB - Gender differences have been observed in a number of aspects of the pharmacology of opiates, including their antinociceptive activity, discriminative stimulus properties, the generation of physical dependence, and their positive reinforcing properties. The current experiments were carried out to rigorously examine whether gender differences exist in the intravenous (IV) self-administration of opiates in an operant conditioning paradigm. Both dose-response analyses and the determination of the strength of the reinforcing properties of opiates using a "breakpoint" analysis were examined. We found strong gender differences in the IV self-administration of two mu opiate agonists-heroin and morphine. At a standard fixed ratio (FR) of responding, females consumed significantly greater amounts of heroin and morphine than did males in a dose-dependent fashion. In addition, females also showed much higher breakpoints than did males: the highest FR breakpoint achieved in females was more than double that observed in males and the frequency distribution of breakpoints was shifted significantly to the right in females when compared to males. These data collectively show that mu opiate agonists may serve as reinforcing agents in females over a broader dose range than males and that they also self-administer considerably more opiates on a milligram per kilogram basis. Finally, we conclude that they will also expend much greater effort in an operant conditioning task to obtain opiate reinforcement. PMID- 12543218 TI - Nitroglycerin inhibits the development of morphine tolerance and dependence in rats. AB - The development of tolerance to and physical dependence on opioids remains a significant barrier to their clinical use. N-Methyl-D-aspartate (NMDA) receptor antagonists inhibit tolerance and dependence. However, many NMDA antagonists have undesirable side effects. It has been shown that nitroglycerin (NTG) can antagonize NMDA receptor activity. This study was designed to determine whether NTG could inhibit the development of morphine tolerance and dependence. Rats were anesthetized and implanted with either morphine or placebo pellets, and pumps infusing vehicle or NTG (doses from 0.1 microg/kg/day to 10 mg/kg/day). Tolerance development was assessed by tail-flick latency (TFL). After 6 days, withdrawal was precipitated by subcutaneous injection of 2 mg/kg naloxone. Withdrawal signs were observed for 15 min. Placebo-pelleted rats showed no changes in TFL over the course of the study and no withdrawal signs. Morphine-pelleted rats developed tolerance. The 0.1 mg/kg/day NTG dose significantly attenuated tolerance development, while the other doses had no significant effect. The 0.1 mg/kg/day dose also attenuated some withdrawal signs. Higher or lower doses were not effective, possibly because of competing biochemical effects. PMID- 12543219 TI - 5-HT1A receptor activity disrupts spontaneous alternation behavior in rats. AB - Agonists selective for three different serotonin (5-HT) receptor subtypes were tested for the ability to disrupt spontaneous alternation behavior (SAB) in the CD strain of rats. Rats were scored for alternation or repetition in their choice of arms of a T-maze equally baited with chocolate milk. Compared with vehicle controls, the 5-HT(1A) agonist 8-hydroxy-dipropylaminotetraline (8-OH-DPAT; 2 mg/kg) significantly (P<.0001) increased repetitive choices (disrupted SAB). In contrast, intraperitoneal injections with the 5-HT(2) agonist R-(-) dimethoxyiodophenylaminoethane (DOI; 1 mg/kg) or the 5-HT(3) agonist N-methyl quipazine (NMQ; 3 mg/kg) had no significant effect on SAB in CD rats. Onset of vicarious trial and error (VTE) behavior prolonged the time required for each rat to select an arm of the T-maze when injected with either 8-OH-DPAT (P<.0001) or buspirone (1-2 mg/kg), a 5-HT(1A) partial agonist. The disruption of SAB and the induction of VTE behavior were reversible with behavioral scores returning to preinjection levels within 48 h after injections. The disruption of SAB by 8-OH DPAT was also seen with the Long-Evans rat strain. The results extend the use of the SAB model and point to a specific role of 5-HT(1A) receptors in the induction of repetitive behavioral patterns. PMID- 12543220 TI - Coadministration of gabapentin or MK-801 with lamotrigine slows tolerance to its anticonvulsant effects on kindled seizures. AB - The development of tolerance to therapeutic effects of antiepileptic drugs can be a problem in the treatment of epilepsy, bipolar disorder, and pain syndromes. In the present study, acute treatment with the new antiepileptic drug lamotrigine (LTG, 15 mg/kg) markedly suppressed seizure stage and seizure duration in amygdala-kindled rats; but this antiseizure effect was rapidly lost following 4-8 days of repeated treatment. When gabapentin (GBP, 20 mg/kg) was coadministered with LTG, the ability of LTG to suppress seizure stage, seizure duration, and after-discharge (AD) duration was markedly extended. In addition, GBP coadministration with LTG decreased the number of animals that developed LTG related running fits (Stage 6 seizures) and lengthened the number of days required to develop running fits or complete tolerance. Neither acute nor repeated treatment with MK-801 (0.3 mg/kg), a noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist, had effects on kindled seizures. However, cotreatment with MK-801 markedly extended the anticonvulsant effects of LTG on the three seizure indices and reduced running fits. These data indicate that cotreatment with either GBP or MK-801 slows tolerance development to the anticonvulsant effects of LTG on kindled seizures. Therapeutic implications of the present study remain to be explored. PMID- 12543221 TI - The GABA(A) receptor mediates the hypnotic activity of melatonin in rats. AB - The present investigation assessed whether hypnotic activity of melatonin was mediated by the GABA(A) receptor in rats. Electroencephalography (EEG) was measured in this experiment. Melatonin, at a dose of 10 mg/kg ip, showed a significant sleep-promoting effect in rats. Flumazenil (3.5 and 7 mg/kg), a specific antagonist of the benzodiazepine (BZP) recognition site on the GABA(A) receptor, and picrotoxin (2 and 4 mg/kg), the ligand of the picrotoxin site on the GABA(A) receptor, seemed to be devoid of intrinsic influence on each sleep parameter when used alone, but they significantly antagonized the melatonin induced increase in total sleep time (TS), slow-wave sleep time (SWS) and paradoxical sleep time (PS), and the decrease in time to sleep onset (TSO) and wakefulness time (W). A significant interaction was shown between melatonin and flumazenil or picrotoxin. When bicuculline methiodide (2 and 4 mg/kg), a specific antagonist of the GABA binding site on the GABA(A) receptor, was used together with melatonin, the melatonin-induced increase in TS, SWS and PS, and the decrease in W were abolished. However, there was no interaction between melatonin and bicuculline methiodide on sleep parameters except PS. These results indicate that the hypnotic activity of melatonin may be linked to the GABA(A) receptor and mediated through the BZP recognition site, the picrotoxin site on the GABA(A) receptor and partially through the GABA binding site on the GABA(A) receptor. PMID- 12543222 TI - Effect of Centella asiatica on pentylenetetrazole-induced kindling, cognition and oxidative stress in rats. AB - Cognitive impairment in epileptics may be a consequence of the epileptogenic process as well as antiepileptic medication. Thus, there is a need for drugs, which can suppress epileptogenesis as well as prevent cognitive impairment. In the present study, the effect of aqueous extract of Centella asiatica (CA) (100 and 300 mg/kg), an Indian medicinal plant known to possess antiepileptic, cognitive-enhancing and antioxidant property, was evaluated on the course of kindling development, kindling-induced learning deficit and oxidative stress markers in pentylenetetrazole (PTZ) kindled rats. Male Wistar rats were injected PTZ (30 mg/kg ip) once every alternate day (48+/-2 h) until the development of the kindling. Passive avoidance test and spontaneous locomotor activity were carried out 24 and 48 h after the last administration of PTZ, while the oxidative stress parameters (malondialdehyde [MDA] and glutathione) were carried out in the whole brain upon completion of the behavioral assessment. The administration of CA (300 mg/kg orally) decreased the PTZ-kindled seizures and showed improvement in the learning deficit induced by PTZ kindling as evidenced by decreased seizure score and increased latencies in passive avoidance behavior. However, low dose of the CA (100 mg/kg) showed improvement only in the learning deficit due to the kindling and failed to improve the seizure score. The findings suggest the potential of aqueous extract of CA as adjuvant to antiepileptic drugs with an added advantage of preventing cognitive impairment. PMID- 12543223 TI - Behavioural and microdialysis study after neurotoxic lesion of the dorsal raphe nucleus in rats. AB - The study investigated the effects of a 5,7-dihydroxytryptamine (5,7-DHT) lesion of the dorsal raphe nucleus (DRN) on anxiety-related behaviour and neurochemical correlates in rats. Behaviour was assessed in the elevated plus maze test (X maze). Lesion of the DRN reduced markedly 5-HT levels in projection areas by at least 60%. Destruction of the serotonergic neurons in the DRN changed neither anxiety-related behaviour on the elevated plus maze, nor aversion-induced 5-HT release in the brain. Exposure of the lesioned rats to the elevated plus maze increased extracellular 5-HT (148%) in the ventral hippocampus similar as in sham lesioned (162%) and non-lesioned (160%) controls. The results demonstrate that lesioning of 5-HT neurons in the DRN does not abolish totally the control of anxiety-related behaviour. PMID- 12543224 TI - Comparative anticonvulsant activity of some 2,3-benzodiazepine derivatives in rodents. AB - The anticonvulsant activities of some 2-amino-3-(3-hydroxy-5-methylisoxazol-4 yl)propionic acid (AMPA)/kainate receptor antagonists, noncompetitive (2,3 benzodiazepines) and a competitive 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F) quinoxaline (NBQX), were compared in different experimental seizure models. In particular, compounds were evaluated against audiogenic seizure in DBA/2 mice, maximal electroshock seizure (MES) test and various chemoconvulsant models; both groups showed a protective action against audiogenic seizure, MES- and pentylenetetrazole (PTZ)-induced seizures. All 2,3-benzodiazepines were also protective against clonic and tonic seizures and lethality induced by 4 aminopyridine, kainate, AMPA and 3-mercaptopropionic acid but were ineffective against NMDA-induced seizures. NBQX was unable to affect 4-aminopyridine-, mercaptopropionic acid- and NMDA-induced seizures. The duration of anticonvulsant action of 33 micromol/kg of some 2,3-benzodiazepine in DBA/2 mice, genetically susceptible to audiogenic seizures, was also investigated. The derivatives possessing a thiocarbonyl group at the C-4 position of heptatomic ring showed higher anticonvulsant activities and longer lasting protective effects. We conclude that all 2,3-benzodiazepines studied are effective against various models of experimental epilepsy and the presence of thiocarbonyl groups at the C 4 position of heptatomic ring is able to increase the anticonvulsant effect of these compounds. PMID- 12543225 TI - Atropine reverses the antinociception of nonsteroidal anti-inflammatory drugs in the tail-flick test of mice. AB - The nonsteroidal anti-inflammatory drugs (NSAIDs) clonixin, diclofenac, piroxicam, ketoprofen, meloxicam, and paracetamol induced antinociception after intraperitoneal or intrathecal administration in mice submitted to an acute thermal algesiometric test without inflammation (tail-flick). Antinociception was evaluated by the increase in reaction time difference (Delta latency), between readings obtained before and after the administration of drugs. The antinociception induced by doses of NSAIDs producing between 20% and 30% of the maximum possible effect (MPE) 30 min after intraperitoneal and 15 min after intrathecal injections was compared with the antinociception obtained after pretreatment with 1 mg/kg atropine ip, 30 min before. Systemic atropine (1 mg/kg) significantly antagonized NSAID-induced antinociception in all cases, both after intraperitoneal and intrathecal administration. Cholinergic depletion by intracerebroventricular hemicholinium-3 (HC-3, 5 microg) 5 h before prevented the antinociceptive effect of all NSAIDs. These observations suggest that intrinsic muscarinic cholinergic facilitatory pathways represent an important modulating system in pain perception in this animal model of acute thermal pain. The results of the present work support the increasingly accepted notion that NSAIDs are effective analgesics even when inflammation is not present, acting by mechanisms that involve actions on spinal and supraspinal nociceptive transmission. It is suggested that, similar to morphine and clonidine, the active mechanism of NSAIDs may involve the release of acetylcholine (ACh) in the spinal cord. PMID- 12543226 TI - Rapid habituation of hippocampal serotonin and norepinephrine release and anxiety related behaviors, but not plasma corticosterone levels, to repeated footshock stress in rats. AB - Prior stress exposure is known to alter the activation response to a subsequent stressor. In the present study, we examined neurochemical, neuroendocrinological, and behavioral correlates of short-term adaptation to homotypic stressors administered 60 min apart. An initial electric footshock significantly induced extracellular levels of both serotonin (5-HT) and norepinephrine (NE) in the rat hippocampus (650% and 200% above baseline, respectively), as measured by in vivo microdialysis. A rapid habituation in this response was evident in the inability of a second footshock to evoke similar increases. In contrast, the hypothalamic pituitary-adrenal (HPA) response was augmented further after the second shock session: plasma corticosterone (CORT) levels were 18.1, 316.5, and 441.6 mg/ml in nonstressed, one-footshock-, or two-footshock-treated rats, respectively. In a social interaction paradigm, rats subjected to a single footshock showed several fear- and anxiety-related behaviors such as increases in freezing and decreases in rearing and active approach for social interaction. Exposure to a second footshock completely blocked the freezing response and restored rearing behavior without affecting the disruption in social interactions. Taken together, these data raise the possibility that neurochemical and neuroendocrine adaptations to short-term homotypic stressors differentially contribute to expression of different fear and anxiety-like responses in the rat. PMID- 12543227 TI - Effects of beta-funaltrexamine and naloxonazine on single-trial morphine conditioned place preference and locomotor activity. AB - The current study assessed the ability of the selective irreversible mu-opioid receptor antagonists beta-funaltrexamine (betaFNA) and naloxonazine (NALZ) to alter the locomotor and rewarding effects of a single intravenous injection of morphine using the conditioned place preference (CPP) model. In the first experiment, rats were conditioned with a single injection of morphine (10 mg/kg iv) paired with one compartment of a CPP apparatus and then were tested for CPP at either 1 or 7 days after conditioning. Rats showed hypoactivity following acute morphine on the conditioning trial and showed CPP when tested either 1 or 7 days later. In the next experiments, rats were pretreated with betaFNA (20 mg/kg sc, 20 h before conditioning), NALZ (15 or 30 mg/kg sc, 24 h before conditioning) or saline and then were conditioned with a single injection of morphine (10 mg/kg iv) or saline. Pretreatment with NALZ alone, but not betaFNA, significantly decreased locomotor activity; neither antagonist alone produced a significant shift in preference for either compartment of the CPP apparatus. Pretreatment with either betaFNA or NALZ blocked completely morphine-induced hypoactivity, but neither antagonist had a significant effect on morphine CPP. These results indicate that mu-opioid receptors are more critically involved in acute morphine induced hypoactivity than in acute morphine reward. PMID- 12543228 TI - Conditioned place aversion and self-administration of nitrous oxide in rats. AB - The rewarding/aversive effects of nitrous oxide (N(2)O) were evaluated using the place conditioning paradigm. Male Long-Evans rats (N=103) received a daily 40-min gas exposure for 8 consecutive days that alternated between two distinct chambers. A control group received placebo gas in both chamber types, while the N(2)O groups (8%, 15%, 30%, and 60% N(2)O) received four consistent pairings of N(2)O with one chamber type and four pairings of placebo gas with the other. A conditioned place aversion was found for the chambers that had been paired with 30% and 60% N(2)O. Place aversions were demonstrated during a 20-min test session on Day 9 when placebo gas was delivered to both chambers, and also during a 20 min test session on Day 10 when N(2)O was delivered to both chambers. A second study evaluated two novel methods of inhalant self-administration, one that used a forced-choice alternating gas environment and one that used a free-choice paradigm. Of four rats tested, two self-administered N(2)O, one rat avoided N(2)O, and one rat's behavior was consistent with neither self-administration nor avoidance. Availability of these methods will facilitate research on the neurobiological mechanisms underlying the rewarding and reinforcing effects of N(2)O and other abused inhalants. PMID- 12543229 TI - Amphetamine-sensitized rats show sugar-induced hyperactivity (cross sensitization) and sugar hyperphagia. AB - The goal was to determine the locomotor and consummatory effects of sugar in amphetamine-sensitized rats. Following a 30-min locomotor activity baseline using a photocell cage, male rats were administered either 3.0 mg/kg amphetamine or saline i.p. daily for 6 days. On the final day of injections, locomotor activity was measured again to affirm amphetamine sensitization. Experiment 1: Seven days later, half of each group was offered 10% sucrose or water for 1 min in the home cages, followed by a 30-min locomotor activity test to determine whether or not the animals had become hyperactive in response to sugar. Results showed that amphetamine-sensitized animals were hyperactive following a taste of sugar, but not water. Experiment 2: All subjects were then given access to 10% sucrose for 1 h daily for five consecutive days. Results showed that the amphetamine-sensitized group consumed more sucrose across the 5-day measurement period. These results suggest that sugar may be acting on the same system as amphetamine to trigger hyperactivity, and that alterations in this system caused by repeated doses of amphetamine can instigate an appetite for sugar that persists for at least a week. PMID- 12543230 TI - Possible mechanism of action in melatonin attenuation of haloperidol-induced orofacial dyskinesia. AB - Tardive dyskinesia (TD) is a late complication of prolonged neuroleptic treatment characterized by involuntary movements of the oral region. In spite of high incidence and much research, the pathophysiology of this devastating movement disorder remains elusive. Chronic treatment with neuroleptics leads to the development of abnormal oral movements in rats, referred to as vacuous chewing movements (VCMs). VCMs in rats are widely accepted as an animal model of TD. Rats chronically treated with haloperidol (1.5 mg/kg ip) significantly developed VCMs and tongue protrusions. Melatonin dose-dependently (1, 2, and 5 mg/kg) reversed the haloperidol-induced VCM and tongue protrusions frequencies. Biochemical analysis reveals that chronic haloperidol treatment significantly induced lipid peroxidation and decreased the forebrain glutathione (GSH) levels in the rats. Chronic haloperidol-treated rats also showed decreased levels of antioxidant defense enzymes, superoxide dismutase (SOD), and catalase. Coadministration of melatonin (1, 2, and 5 mg/kg) along with haloperidol significantly reduced the lipid peroxidation and restored the decreased GSH levels by chronic haloperidol treatment, and significantly reversed the haloperidol-induced decrease in forebrain SOD and catalase levels in rats. However, a lower dose of melatonin (1 mg/kg) failed to reverse chronic haloperidol-induced decreases in forebrain GSH, SOD, and catalase levels. In conclusion, melatonin could be screened as a potential drug candidate for the prevention or treatment of neuroleptic-induced orofacial dyskinesia. PMID- 12543231 TI - Involvement of the kappa-opioid receptor in the anxiogenic-like effect of CP 55,940 in male rats. AB - We have studied the possible interaction between three selective opioid-receptor antagonists, nor-binaltorphimine (NB: kappa) (5 mg/kg), cyprodime (CY: mu) (10 mg/kg) and naltrindole (NTI: delta) (1 mg/kg), and the cannabinoid receptor agonist CP 55,940, in the modulation of anxiety (plus-maze) and adrenocortical activity (serum corticosterone levels by radioimmunoassay) in male rats. The holeboard was used to evaluate motor activity and directed exploration. CP 55,940 (75 microg/kg, but not 10 microg/kg) induced an anxiogenic-like effect, which was antagonised by NB. The other effects of CP 55,940 (75 microg/kg), a decreased holeboard activity and stimulation of adrenocortical activity, were not antagonised by any of the three opioid receptor antagonists. CY and NTI, when administered alone, induced marked reductions in motor activity, anxiogenic-like effects and stimulation of adrenocortical activity. The selective kappa-opioid receptor antagonist NB, on its own, did not modify the level of anxiety but stimulated adrenocortical activity. We provide the first pharmacological evidence about the involvement of the kappa-opioid receptor in the anxiogenic-like effect of CP 55,940. PMID- 12543232 TI - Spinal nociceptin inhibits AMPA-induced nociceptive behavior and Fos expression in rat spinal cord. AB - The effects of intrathecal nociceptin (NOCI) on the nociceptive behavior (biting, scratching and licking; BSL) and the spinal Fos expression induced by intrathecal administration of N-methyl-D-aspartate (NMDA, 4 microg/rat) or alpha-amino-3 hydroxy-5-methylisoxazole-4-propionate (AMPA, 2 microg/rat) were studied. Coadministration of NOCI (3 and 10 nmol/rat) with NMDA did not modify the NMDA induced BSL or Fos expression. In contrast, NOCI (0.1-3 nmol/rat) dose dependently inhibited the BSL behavior induced by AMPA. Furthermore, coadministration of NOCI (3 and 10 nmol/rat) significantly reduced the AMPA induced expression of Fos protein in the superficial layers of the spinal dorsal horn. In order to test whether classical or opioid receptor-like type 1 (ORL1) receptors are involved in the inhibitions by NOCI of AMPA-evoked BSL, the corresponding antagonists were assayed. The administration of the nonselective opioid receptor antagonist, naloxone (10 mg/kg i.p.), did not modify the NOCI induced inhibition of AMPA-evoked BSL. However, the selective ORL1 receptor antagonist, [N-Phe(1)]nociceptin-(1-13)-NH(2) (90 nmol/rat i.t.), completely prevented the NOCI-mediated inhibition of the nociceptive responses evoked by AMPA. In conclusion, NOCI, acting at ORL1 receptors can, at least in part, induce spinal analgesia by blocking the nociceptive responses produced through the stimulation of AMPA receptors. PMID- 12543233 TI - Forelimb use after focal cerebral ischemia in rats treated with an alpha 2 adrenoceptor antagonist. AB - Atipamezole, a selective alpha(2)-adrenoceptor antagonist, enhances recovery of sensorimotor function after focal cerebral ischemia in rats. The aim of the present study was to further characterize the effects of atipamezole treatment combined with enriched-environment housing in ischemic rats by evaluating spontaneous exploratory activity in the cylinder test. The right middle cerebral artery (MCA) of rats was occluded for 120 min using the intraluminal filament method. Atipamezole (1.0 mg/kg) or 0.9% NaCl was administered on postoperative days 2 through 11 and 15, 19, and 23. Spontaneous behavior of rats in a transparent cylinder was videotaped before, and 6 and 23 days after surgery 20 min after drug administration. Constant asymmetry in forelimb use was observed in the cylinder test on postoperative days 6 and 23. Ischemic rats used the impaired forelimbs (contralateral to lesion) during lateral exploration less than did sham operated rats (P<.001). Ischemic rats also preferred to turn contralateral to the lesion (P<.05). Atipamezole increased the simultaneous, but not independent, use of the forelimbs during lateral exploration (P<.05). The data suggest that noradrenergic manipulation does not significantly enhance recovery in a test that does not depend on practice following focal cerebral ischemia. PMID- 12543234 TI - Prolonged perinatal AZT administration and early maternal separation: effects on social and emotional behaviour of periadolescent mice. AB - Zidovudine (AZT) is an effective treatment in preventing perinatal transmission of HIV-1; however, a continuous re-evaluation of the risk-benefit ratio of human exposure to this drug is suggested by both clinical and animal studies. The objective of this study was to assess the medium and long-term effects of pre postnatal AZT treatment on mouse social and emotional behaviour and the possible interactions between AZT exposure and disruptions in the mother-infant relationship. Pregnant CD-1 mice were administered per os with AZT (160 mg/kg) from pregnancy day 10, throughout delivery, to lactation day 10. In half of the litters, the offspring was separated from the mother for 3 h from postnatal days 2 (PND2) to PND14. On PND35, a 30-min social interaction test was performed and corticosterone levels were measured at the end of the session. On PND80, long term effects of AZT on emotionality were assess by means of an elevated plus maze. Results indicate that, on PND35, previous AZT exposure affected social behaviour of the experimental subjects, reducing aggressive interactions in males, while decreasing investigative behaviours in females. At adulthood, AZT inhibited exploratory behaviour in the plus-maze while increasing the frequency of risk-assessment postures in male mice. As for maternal deprivation, this early manipulation exerted a pro-aggressive effect in adolescent male mice, deprived subjects being overall characterised by higher activity levels and a deficit in habituation, an effect also observed in the plus-maze. A significant interaction between AZT and maternal deprivation was found for affiliative behaviours. As for corticosterone levels, no AZT effect was found, while maternal deprivation tended to reduce elevations of this hormone in response to stressful stimuli. Overall results from this study indicate that both AZT exposure and maternal deprivation induced gender-dependent changes in social and emotional behaviour both during adolescence and at adulthood. PMID- 12543235 TI - Nicotine-induced changes in cerebrocortical neuroactive steroids and plasma corticosterone concentrations in the rat. AB - Nicotine, one of the most widely used psychotropic substances, is able to induce both anxiolytic and anxiogenic effects. The effect of this drug on the brain and plasma concentrations of neuroactive steroids was examined in the rat. Anxiolytic doses of nicotine (0.03-0.3 mg/kg) had no significant effect, whereas administration of anxiogenic doses (0.5 to 2 mg/kg) produced a dose- and time dependent increase in the cerebrocortical concentrations of pregnenolone, progesterone, and allopregnanolone, with the greatest observed effects (+180%, +223%, and +124%, respectively) apparent at the dose of 2 mg/kg. In contrast, nicotine (1-2 mg/kg) decrease by 31% and 38%, respectively, the concentration of 3alpha,21-dihydroxy-5alpha-pregnan-20-one (allotetrahydrodeoxycorticosterone, or THDOC) in the cerebral cortex. Nicotine also increased the plasma concentrations of pregnenolone and progesterone, whereas failed to affect significantly those of allopregnanolone or THDOC. Nicotine induced a dose- and time-dependent increase in the plasma concentration of corticosterone, indicating that this drug activates the hypothalamic-pituitary-adrenal (HPA) axis. These results suggest that the changes in emotional behavior elicited by nicotine, similar to those induced by stressful stimuli or other anxiogenic drugs, are associated with an increase in neuroactive steroids content of the brain. PMID- 12543236 TI - The effect of electroacupuncture as an adjunct on cyclophosphamide-induced emesis in ferrets. AB - The effect of electroacupuncture (EA) on cyclophosphamide-induced emesis in ferrets was studied at acupuncture point Neiguan (P6) with various electrical stimulation parameters (5-100 Hz, 1.5-3 V, 5-20 min, n=6/group). The combination therapy of EA (100 Hz, 1.5 V and 10 min) with the lower doses of ondansetron (0.04 mg/kg), droperidol (0.25 mg/kg) and metoclopramide (2.24 mg/kg) significantly reduced the total number of emetic episodes by 52%, 36% and 73%, respectively, as well as the number of emetic episodes in the first phase as compared to the sham acupuncture control (P<.01). These EA/drug combinations also showed a significant effect in preventing emesis as compared to either EA or drug alone (P<.05). The present study suggests that acupuncture may be useful as an adjunctive therapy in the treatment of chemotherapy-induced emesis. PMID- 12543237 TI - Cocaine-conditioned behavioral effects: a role for habituation processes. AB - Cocaine has potent locomotor stimulant effects in rodents, which seemingly can become conditioned to test environment cues. In two experimental protocols, we measured the effects of cocaine on locomotor activity and grooming behavior, and subsequently tested whether these cocaine effects became conditioned to contextual cues. In the first experiment, three groups of rats received 14 injections of either saline or cocaine (10 mg/kg) paired or unpaired to the test environment. Cocaine increased locomotion and decreased grooming during treatment and on the conditioning test. Over the course of the treatment phase, however, the saline- and cocaine-unpaired groups but not the cocaine paired group developed progressively lower locomotion and higher grooming scores indicative of substantial habituation effects. To examine whether the cocaine may have impaired the acquisition of habituation effects rather than induce a Pavlovian cocaine conditioned response, an additional experiment was conducted in which two additional non-habituation saline and cocaine control groups were added to the experimental design. On a conditioning test, the two non-habituation control groups were equivalent in activity and grooming behavior to the cocaine-paired group. The findings were consistent with a failure by cocaine-paired animals to acquire habituation effects, which could transfer to the non-cocaine state. The connection between cocaine and novelty/habituation may have substantial importance for understanding cocaine effects. PMID- 12543238 TI - Characterization of the discriminative stimulus properties of centrally administered (-)-DOM and LSD. AB - Despite the plausible assumption that the effects of hallucinogens predominantly arise in the central nervous system, most studies of these drugs in intact subjects have been conducted following systemic administration. The objective of the present investigation was to characterize the stimulus effects of (-)2,5 dimethoxy-4-methylamphetamine ((-)-DOM) following intracerebroventricular administration. Chronic indwelling cannulae were implanted into the lateral ventricle of male Fischer 344 rats trained to discriminate systemically administered (-)-DOM or lysergic acid diethylamide (LSD) from saline. Time-course and dose-response relationships for (-)-DOM and LSD administered intracerebroventricularly were established. For both LSD and (-)-DOM, central administration did not change the pretreatment times required for the maximal stimulus effects to occur. However, the onset of the stimulus effect was more rapid following intracerebroventricular administration. Following pretreatment periods that maximize drug-appropriate responding, central administration of (-) DOM and LSD was approximately 2.4- and 1.5-times more potent, respectively, than systemic administration. The results of this study are consistent with the assumption that the stimulus effects of (-)-DOM and LSD are centrally mediated. PMID- 12543239 TI - Smoked heroin in rhesus monkeys: effects of heroin extinction and fluid availability on measures of heroin seeking. AB - The purpose of the present study was to evaluate the reinforcing effects of smoked heroin in nonopioid-dependent nonhuman primates when an alternative reinforcer, sweetened fluid, was made available. Four adult male rhesus monkeys lived in three chambers, with heroin self-administration (0, 0.3, and 0.6 mg/kg) specific to one end of the chamber, oral sweetened fluid self-administration specific to the other end chamber, and no commodity available in the middle chamber. The length of time monkeys spent in the drug-associated chamber provided one measure of drug seeking (i.e., location preference). During self administration sessions, a second-order schedule of reinforcement was used, with responding during the first component maintained by a brief presentation of the stimuli associated with reinforcement. Responding during the second component was maintained by a delivery of the reinforcer, and the associated stimuli. Responding during the first component provided a second measure of drug seeking. Monkeys also had choice trials each day, when they could choose to work for either commodity. Choice behavior provided a third measure of drug seeking. Each experimental day consisted of a smoking session (four smoking trials), a sweetened fluid session (four fluid trials), and a choice session (four choice trials). Monkeys typically completed all four smoking trials each day when either of the active heroin doses was available. They chose both heroin doses over fluid on 3.5 of the four choice trials, and they had a location preference for the heroin chamber. Under baseline conditions, the number of acquisition responses and the number of consumption responses (inhalations) were greater for the high dose of heroin compared to the low dose of heroin. Further, it took longer to extinguish the responding for the high dose of heroin compared to the low dose of heroin when a vehicle was substituted. During heroin extinction, acquisition responding for fluid increased, the number of fluid choices increased, and location preference shifted to the fluid chamber. These data suggest that in nondependent rhesus monkeys, measures of heroin seeking decreased when heroin was not available and seeking behavior shifted to the available alternative commodity. PMID- 12543240 TI - Clove cigarette smoking: biochemical, physiological, and subjective effects. AB - Alternative tobacco products such as clove (kreteks) and bidi cigarettes have become increasingly popular among US smokers. The nicotine content of a popular clove cigarette (Djarum Special) filler averaged 7.4 mg; conventional cigarettes contained 13.0 mg. However, smoke yields from standardized machine-smoking analysis indicated it delivered more nicotine, carbon monoxide (CO), and tar than conventional cigarettes. In a clinical study, nicotine delivery, physiologic, and subjective effects of the clove cigarette were compared to their own brand of cigarette in 10 adult smokers (7 males). Average time to smoke the clove cigarette (549 s) and number of puffs (15.1) were significantly greater than own brand (314 s and 9.4 puffs). Increases in venous plasma nicotine and exhaled CO after smoking the clove cigarette (17.4 ng/ml; 6 ppm) were similar to those after own brand (17.6 ng/ml; 4.5 ppm). Maximal changes in heart rate (HR), systolic, and diastolic blood pressures (BP) did not differ significantly between the clove and own brand of cigarette. Compared to their own brand of cigarette, the clove cigarette was rated as better tasting and being distinctly different. Our findings indicate that clove cigarettes deliver significant quantities of nicotine, CO, and presumably other toxic components of tobacco smoke. Taste satisfaction, aromatic odor, and novelty may contribute to their appeal to young smokers. PMID- 12543241 TI - Neurobehavioral effects of racemic threo-methylphenidate and its D and L enantiomers in rats. AB - D,L-methylphenidate (Ritalin) is used to treat attention deficit hyperactivity disorder (ADHD) in children. The therapeutic effect is predominantly due to the d enantiomer. Dexmethylphenidate (D-MPH; Focalin) was therefore developed for its better therapeutic index. The present study determined and compared the acute behavioral toxicity of D,L-MPH, D-MPH and L-MPH in rats after oral dosing. Comprehensive functional observational battery (FOB) evaluations and rota-rod tests were performed 30, 60 and 120 min after dosing. Ten rats/sex/dose were administered a single dose of vehicle, 2, 20, 100 mg/kg D,L-MPH and 1, 10, 50 mg/kg D-MPH or 1, 100, 500 mg/kg L-MPH. There was no mortality. Certain FOB evaluations were statistically significant from vehicle control at any of the time points with most occurring at 60 and 120 min in the high D,L-MPH dose. These included increases in rearing, difficulty in removal from box, arousal, click, tail-pinch and decreases in hind-limb splay distance, hind-limb grip strength and handling reactivity. Behavioral responses were also present at the mid-dose D,L MPH and high dose D- and L-MPH. Responses in female were significantly different from males in D,L- and L-MPH groups suggesting a sex difference in sensitivity. In the rota-rod test, mean latency to remain on the rod was significantly less for males compared to control given high dose D-MPH and D,L-MPH. In females, latency times were significantly less for high doses of all three compounds. In summary, fewer significant FOBs were seen with D- and L-MPH compared to equimolar doses of D,L-MPH. L-MPH was the least potent in producing FOBs. These results were supported by rota-rod studies. PMID- 12543242 TI - Strain-dependent differences in schedule-induced polydipsia: an assessment in Lewis and Fischer rats. AB - Strain-dependent differences have been used to highlight unknown genetic contributions to important behavioral and physiological end points. In this regard, the Fischer (F344) and Lewis (LEW) rat strains have often been studied because they exhibit a myriad of behavioral and physiological differences. Recently, schedule-induced polydipsia (SIP), a potential model of stress and drug abuse, has been reported to differ between the two strains (see [Pharmacol. Biochem. Behav. 67 (2002) 809]) with F344 rats displaying greater levels of consumption than LEW rats. Given the importance of SIP as a behavioral model of stress and of drug abuse, the present study further explored SIP in F344 and LEW strains by assessing the acquisition and steady-state performance of SIP (under a fixed-time 30 schedule of food delivery; FT30), its characteristic postprandial temporal licking pattern and its modulation by variations in the food delivery schedule (FT15, FT30 and FT60). F344 rats acquired SIP at a faster rate and drank at a higher asymptotic level than LEW rats. Both strains displayed the typical inverted U-shaped post-pellet pattern of drinking and changes in levels of consumption (and displacement of the initiation of post-pellet drinking) with changes in the FT value, supporting the position that the increased drinking seen in both groups was schedule induced. These strain differences in SIP are consistent with the fact that the F344 and LEW strains differ on other behavioral and physiological indices of stress and raise the issue of the use of this model in the assessment of differential drug intake between the two strains. PMID- 12543243 TI - Effects of diazepam and flumazenil on food competition behavior in high- and low aggression pigeons. AB - The food competition interaction test performed with food-restricted pigeons with previously consolidated dominance is a useful tool for the study of offensive and defensive social aggression. In the present study, we examined the effect of GABA A-benzodiazepine (BZD) receptor manipulation on aggression, emotion, feeder control, and eating behavior in high- and low-aggression female pigeons maintained at 80% of their normal weight and exposed to food competition interactions. The pigeons were divided into pairs by previously ranked high aggression females (total time spent in aggression over 60 s/5 min; n=6 pairs) and low-aggression females (time spent in aggression less than 10 s/5 min; n=6 pairs). In Experiment 1, a pigeon in each pair of high- and low-aggression subjects were treated daily with an oral dose of diazepam (DZP, 0.6 mg/kg/0.3 ml) for 8 days. The other animal received the vehicle. On Day 8, food competition trials (10 min) were performed 30 min after treatments. In Experiment 2, pigeons were injected subcutaneously with flumazenil (FZL, 0.1 mg/kg/1 ml) or saline and exposed to a food competition trial 30 min after injections. In Experiment 3, one animal in each pair received DZP for 8 days. The other animal received the vehicle. On Day 8, the DZP-treated subjects were injected subcutaneously with FZL (0.1 mg/ kg/1 ml) 30 min before the oral dose of DZP. Trials were performed 30 min after DZP or vehicle administration. In Experiment 1, it was found that the DZP group of high-aggression pigeons showed lower scores of aggression (P<.05) and emotional responses (P<.05) than controls. The other group-scored behaviors were not affected. The DZP low-aggressions, however, showed scores of aggression eightfold higher than their controls (P<.05) but the other scored behaviors were not changed. In Experiment 2, FZL injection did not induce intrinsic effects on aggression either in the high- or in the low-aggression group. Experiment 3 showed that the emotional and aggressive responses to DZP were neutralized by FZL. This shows that GABA-A-BZD receptor mechanisms are implicated in the DZP responses in high- and low-aggression pigeons. PMID- 12543244 TI - An in vivo evaluation of the antiseizure activity and acute neurotoxicity of agmatine. AB - Agmatine, an endogenous cationic amine, exerts a wide range of biological effects, including modulation of glutamate-activated N-methyl-D-aspartate (NMDA) receptor function in the central nervous system (CNS). Since glutamate and the NMDA receptor have been implicated in the initiation and spread of seizure activity, the capacity of agmatine to inhibit seizure spread was evaluated in vivo. Orally administered agmatine (30 mg/kg) protected against maximal electroshock seizure (MES)-induced seizure spread in rats as rapidly as 15 min and for as long as 6 h after administration. Inhibition of MES-induced seizure spread was also observed when agmatine was administered intraperitoneally. Agmatine's antiseizure activity did not appear to be dose-dependent. An in vivo neurotoxicity screen indicated that agmatine was devoid of any acute neurological toxicity at the doses tested. These preliminary data suggest that agmatine has promising anticonvulsant activity. PMID- 12543245 TI - Nitric oxide-induced mitochondrial dysfunction: implications for neurodegeneration. AB - Excessive generation of nitric oxide (NO) has been implicated in the pathogenesis of several neurodegenerative disorders. Damage to the mitochondrial electron transport chain has also been implicated in these disorders. NO and its toxic metabolite peroxynitrite (ONOO(-)) can inhibit the mitochondrial respiratory chain, leading to energy failure and ultimately cell death. There appears to be a differential susceptibility of brain cell types to NO/ONOO(-), which may be influenced by factors including cellular antioxidant status and the ability to maintain energy requirements in the face of marked respiratory chain damage. Although formation of NO/ONOO(-) following cytokine exposure does not affect astrocyte survival, these molecules may diffuse out and cause mitochondrial damage to neighboring NO/ONOO(-)-sensitive cells such as neurons. Evidence suggests that NO/ONOO(-) causes release of neuronal glutamate, leading to glutamate-induced activation of neuronal NO synthase and generation of further damaging species. While neurons appear able to recover from short-term exposure to NO/ONOO(-), extending the period of exposure results in persistent damage to the respiratory chain and cell death ensues. These findings have important implications for acute infection vs. chronic neuroinflammatory disease states. The evidence for NO/ONOO(-)-mediated mitochondrial damage in neurodegenerative disorders is reviewed and potential therapeutic strategies are discussed. PMID- 12543246 TI - Suicidal inactivation of the rabbit 15-lipoxygenase by 15S-HpETE is paralleled by covalent modification of active site peptides. AB - Lipoxygenases (LOXs) are multifunctional enzymes that catalyze the oxygenation of polyunsaturated fatty acids to hydroperoxy derivatives; they also convert hydroperoxy fatty acids to epoxy leukotrienes and other secondary products. LOXs undergo suicidal inactivation but the mechanism of this process is still unclear. We investigated the mechanism of suicidal inactivation of the rabbit 15 lipoxygenase by [1-(14)C]-(15S,5Z,8Z,11Z,13E)-15-hydroperoxyeicosa-5,8,11,13 tetraenoic acid (15-HpETE) and observed covalent modification of the enzyme protein. In contrast, nonlipoxygenase proteins (bovine serum albumin and human gamma-globulin) were not significantly modified. Under the conditions of complete enzyme inactivation we found that 1.3 +/- 0.2 moles (n = 10) of inactivator were bound per mole lipoxygenase, and this value did depend neither on the enzyme/inactivator ratio nor on the duration of the inactivation period. Covalent modification required active enzyme protein and proceeded to a similar extent under aerobic and anaerobic conditions. In contrast, [1-(14)C] (15S,5Z,8Z,11Z,13E)-15-hydroxyeicosa-5,8,11,13-tetraenoic acid (15-HETE), which is no substrate for epoxy-leukotriene formation, did not inactivate the enzyme and protein labeling was minimal. Separation of proteolytic cleavage peptides (Lys-C endoproteinase digestion) by tricine SDS-PAGE and isoelectric focusing in connection with N-terminal amino acid sequencing revealed covalent modification of several active site peptides. These data suggest that 15-lipoxygenase catalyzed conversion of (15S,5Z,8Z,11Z,13E)-15-hydroperoxyeicosa-5,8,11,13 tetraenoic acid to 14,15-epoxy-leukotriene leads to the formation of reactive intermediate(s), which are covalently linked to the active site. Therefore, this protein modification contributes to suicidal inactivation. PMID- 12543247 TI - Inhibition of oral cancer cell growth by adenovirusMnSOD plus BCNU treatment. AB - We hypothesized that inhibitors of peroxide removal, such as BCNU, an indirect inhibitor of glutathione peroxidase (GPx), and 3-amino-1,2,4-triazole (AT), a direct inhibitor of catalase (CAT), should cause toxicity to cancer cells after manganese superoxide dismutase (MnSOD) overexpression due to elevated peroxide levels. In vitro, hamster cheek pouch carcinoma cells (HCPC-1) and human oral squamous carcinoma cells (SCC-25) were infected with various combinations of adenovirus containing MnSOD cDNA (AdMnSOD). Cells were then treated with or without BCNU and assayed for viability using Annexin/PI staining and flow cytometry. In AdMnSOD plus BCNU-treated SCC-25 and HCPC-1 cells, a 30-60% decrease in cell viability was observed compared to BCNU alone. In vivo, HCPC-1 and SCC-25 xenografts were allowed to grow to approximately 70 mm(3) and 10(9) plaque forming units (pfu) of AdMnSOD were injected directly into the tumors. Two days later, 15 or 30 mg/kg BCNU was injected intratumorally. Tumor growth was greatly inhibited (4- to 20-fold) by this combined treatment, as well as increasing animal survival. Tumor volume could be decreased further by giving multiple doses of AdMnSOD or inhibiting catalase activity with AT. These results suggest that, by using these combination therapies, a significant decrease in tumor mass can be achieved. PMID- 12543248 TI - HPLC analysis of vitamin E isoforms in human epidermis: correlation with minimal erythema dose and free radical scavenging activity. AB - The content and composition of different vitamin E isoforms was analyzed in normal human skin. Interestingly the epidermis contained 1% alpha-tocotrienol, 3% gamma-tocotrienol, 87% alpha-tocopherol, and 9% gamma-tocopherol. Although the levels of tocotrienol in human epidermis appear to be considerably lower than reported in the hairless mouse, the presence of significant amounts of tocotrienol levels leads to speculation about the physiological function of tocotrienols in skin. Besides antioxidant activity and photoprotection, tocotrienols may have skin barrier and growth-modulating properties. A good correlation was found for epidermal alpha-tocopherol (r = 0.7909, p <.0003), gamma-tocopherol (r = 0.556, p <.025), and the total vitamin E content (r = 0.831, p <.0001) with the free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging in epidermis, as assessed by electron paramagnetic resonance (EPR) spectroscopy. In human epidermis, alpha-tocopherol is quantitatively the most important vitamin E isoform present and comprises the bulk of first line free radical defense in the lipid compartment. Epidermal tocotrienol levels were not correlated with DPPH scavenging activity. The minimal erythema dose (MED), an individual measure for sun sensitivity and a crude indicator for skin cancer susceptibility, did not correlate with the epidermal content of the vitamin E isoforms. Hence it is concluded that vitamin E alone is not a determinant of individual photosensitivity in humans. PMID- 12543249 TI - Chronic ethanol treatment: dolichol and retinol distribution in isolated rat liver cells. AB - The aim of this study was to use chronic ethanol intoxication for 2 and 4 months as a means of studying the distribution of dolichol and retinol in isolated rat liver parenchymal cells, Kupffer cells, sinusoidal endothelial cells, and two subfractions of hepatic stellate cells: Ito 1 and Ito 2. Dolichol and retinol were studied in two batches of rats: on normal nutrition and after a load of vitamin A given 3 d before sacrifice. New observations reported are: (i) on normal nutrition, after 2 months of treatment, dolichol in HC seems to be the first target of chronic ethanol, while retinol is the first target in hepatic stellate cells; (ii) the various types of liver cells are differently affected by chronic ethanol, which highlights the importance of studying each type of sinusoidal cell; (iii) a load of vitamin A given when the damage has already occurred restores dolichol content in HC while retinol decreases; and, (iv) a link between dolichol and vitamin A metabolism might be supposed after the load of vitamin A: the percentage distribution of dolichol with 18 isoprene units (Dolichol -18) increases in all the control cells but decreases after chronic ethanol treatment. A different role of this dolichol and/or a different compartmentalization within the cell need to be further investigated. PMID- 12543250 TI - The cytotoxic effect of volatile organic compounds of the gas phase of cigarette smoke on lung epithelial cells. AB - Health effects of cigarette smoke (CS) in humans are well known from both clinical and epidemiological studies. However, the mechanism behind CS toxicity and carcinogenicity remains mainly unknown. Recent studies have pointed to the major importance of the gas phase of CS in generating its cytotoxic effects. In the current study, an exposure system capable of introducing the gas phase of mainstream cigarette smoke deprived of its volatile organic constituents (VOCs) was used to study the role of the nonorganic components of the gas phase on the cytotoxicity of smoke to monolayer cultures of mouse lung epithelial cells. Cell viability was measured by Wst-1 and the lactate dehydrogenase (LDH) assays. In cells treated with increasing doses of mainstream cigarette smoke gas phase (one to nine puffs), a dose-dependent increase in cytotoxicity was observed (one puff, 95% viability; nine puffs, 40% viability). Cell viability of cultures exposed to gas phase with only the nonorganic components was found to be equivalent to control, unexposed cultures, indicating that removal of VOCs resulted in almost eliminating the cytotoxic ability of the gas phase of CS. Furthermore, the removal of VOCs seems to reduce the effects of protein tyrosine nitration mediated through the gas phase constituents. The results obtained suggest the important and decisive role of VOCs in inducing cytotoxic effects. PMID- 12543251 TI - Phospholipids in oxidized LDL not adducted to apoB are recognized by the CD36 scavenger receptor. AB - Previous studies have shown that oxidation of low-density lipoprotein (oxLDL) results in its recognition by scavenger receptors on macrophages. Whereas blockage of lysyl residues on apoB-100 of oxLDL by lipid peroxidation products appears to be critical for recognition by the scavenger receptor class A (SR-A), modification of the lipid moiety has been suggested to be responsible for recognition by the scavenger class B receptor, CD36. We studied the recognition by scavenger receptors of oxidized LDL in which lysyl residues are blocked prior to oxidation through methylation [ox(m)LDL]. This permits us to minimize any contribution of modified apoB-100 to the recognition of oxLDL, but does not disrupt the native configuration of lipids in the particle. We found that ox(m)LDL was recognized by receptors on mouse peritoneal macrophages (MPM) almost as well as oxLDL. Ox(m)LDL was recognized by CD36-transfected cells but not by SR A-transfected cells. Oxidized phospholipids (oxPC) transferred from oxLDL or directly from oxPC to LDL, conveyed recognition by CD36-transfected cells, confirming that CD36 recognized unbound oxidized phospholipids in ox(m)LDL. Collectively, these results suggest that oxPC not adducted to apoB within the intact oxLDL particle are recognized by the macrophage scavenger receptor CD36, that these lipids are not recognized by SR-A, and that they can transfer from oxidized to unoxidized LDL and induce CD36 recognition. PMID- 12543252 TI - Late preconditioning by ethanol is initiated via an oxidant-dependent signaling pathway. AB - Ingestion of alcoholic beverages at low to moderate levels 24 h prior to ischemia and reperfusion (I/R) prevents postischemic leukocyte/endothelial cell adhesive interactions, a phenomenon referred to as late ethanol preconditioning (EtOH-PC). The aim of this study was to determine whether oxidants act as initiators of late EtOH-PC. Ethanol was instilled into the stomachs of C57BL/6 mice as a bolus by gavage at a dose that produced a peak plasma concentration of 45 mg/dl 30 min after administration and returned to control levels 60 min after ingestion. Twenty four hours later, the superior mesenteric artery was occluded for 45 min followed by 70 min of reperfusion. The numbers of rolling and firmly adherent leukocytes were quantified in postcapillary venules of the small intestine in sham animals (no EtOH-PC, no I/R), in mice subjected to I/R alone or EtOH-PC + I/R, and in animals treated with Mn-TBAP (a cell-permeant superoxide dismutase mimetic), oxypurinol (a XO inhibitor), the NAD(P)H oxidase inhibitors PR-39 or apocynin, or oxypurinol plus PR39 during the period of EtOH-PC on Day 1 followed by I/R on Day 2. In separate groups of mice, oxypurinol or apocynin were also administered 1 h after ethanol ingestion on Day 1, with induction of I/R 24 h later. I/R induced marked increases in leukocyte rolling and adherence, effects that were completely prevented by EtOH-PC. Coincident treatment with Mn-TBAP, oxypurinol, PR-39, apocynin, or oxypurinol plus PR-39 with ethanol attenuated these anti-inflammatory actions of EtOH-PC. However, administration of oxypurinol or apocynin 1 h after ethanol ingestion failed to prevent these protective effects of EtOH-PC. Our results indicate that reactive oxygen species formed during the period of ethanol exposure on Day 1 trigger the development of an anti inflammatory phenotype that renders the small bowel resistant to the proadhesive effects of I/R 24 h later. PMID- 12543253 TI - Inhibition of oral peroxidase activity by cigarette smoke: in vivo and in vitro studies. AB - Oral peroxidase (OPO), the pivotal enzyme in the salivary antioxidant system, seems to be of paramount importance in the oral defense mechanism, especially against the attack of free radicals related to cigarette smoke (CS) and the evolution of oral cancer. The major inducer of oral cancer is exposure to tobacco, which is responsible for 50-90% of cases worldwide. The purpose of our study was to elucidate the outcome of interaction between CS and OPO in smokers and nonsmokers. After smoking a single cigarette, a sharp drop of OPO activity was observed in both groups: 42.5% in smokers and 58.5% in nonsmokers (p <.05). After 30 min, the level of activity returned to 90-100% of the presmoking level, presumably due to the secretion of new saliva into the oral cavity. The difference between the two groups was also observed after exposure of saliva to one cigarette in smoking flasks (in vitro studies); however, as expected, no recovery of activity was observed in either group. Similarly, the OPO activity loss was accompanied by increased carbonylation of the salivary proteins, an indicator of the oxidative damage to proteins. These results may be of great clinical importance, as heavy smokers smoke 20 cigarettes or more on a daily basis. Accordingly, most of the time the oral epithelium of heavy smokers is essentially unprotected by OPO against the deleterious effects of thiocyanate ions and hydroxyl radicals produced by unremoved hydrogen peroxide in the presence of the salivary redox-active metal ions. This may pave the way for the CS-induced and saliva-mediated initiation and progression of oral cancer. PMID- 12543254 TI - Regulation of antioxidant enzymes gene expression in the yeast Saccharomyces cerevisiae during stationary phase. AB - Gene expression of three antioxidant enzymes, Mn superoxide dismutase (MnSOD), Cu,Zn superoxide dismutase (Cu,ZnSOD), and glutathione reductase (GR) was investigated in stationary phase Saccharomyces cerevisiae during menadione induced oxidative stress. Both GR and Cu,ZnSOD mRNA steady state levels increased, reaching a plateau at about 90 min exposure to menadione. GR mRNA induction was higher than that of Cu,ZnSOD (about 14-fold and 9-fold after 90 min, respectively). A different pattern of response was obtained for MnSOD mRNA, with a peak at about 15 min (about 8-fold higher) followed by a decrease to a plateau approximately 4-fold higher than the control value. However, these increased mRNA levels did not result in increased protein levels and activities of these enzymes. Furthermore, exposure to menadione decreased MnSOD activity to half its value, indicating that the enzyme is partially inactivated due to oxidative damage. Cu,ZnSOD protein levels were increased 2-fold, but MnSOD protein levels were unchanged after exposure to menadione in the presence of the proteolysis inhibitor phenylmethylsulfonyl fluoride. These results indicate that the rates of Cu,ZnSOD synthesis and proteolysis are increased, while the rates of MnSOD synthesis and proteolysis are unchanged by exposure to menadione. Also, the translational efficiency for both enzymes is probably decreased, since increases in protein levels when proteolysis is inhibited do not reflect the increases in mRNA levels. Our results indicate that oxidative stress modifies MnSOD, Cu,ZnSOD, and GR gene expression in a complex way, not only at the transcription level but also at the post-transcriptional, translational, and post-translational levels. PMID- 12543256 TI - Cellular senescence and apoptosis: how cellular responses might influence aging phenotypes. AB - Aging in complex multi-cellular organisms such as mammals entails distinctive changes in cells and molecules that ultimately compromise the fitness of adult organisms. These cellular and molecular changes lead to the phenotypes we recognize as aging. This review discusses some of the cellular and molecular changes that occur with age, specifically changes that occur as a result of cellular responses that evolved to ameliorate the inevitable damage that is caused by endogenous and environmental insults. Because the force of natural selection declines with age, it is likely that these processes were never optimized during their evolution to benefit old organisms. That is, some age related changes may be the result of gene activities that were selected for their beneficial effects in young organisms, but the same gene activities may have unselected, deleterious effects in old organisms, a phenomenon termed antagonistic pleiotropy. Two cellular processes, apoptosis and cellular senescence, may be examples of antagonistic pleiotropy. Both processes are essential for the viability and fitness of young organisms, but may contribute to aging phenotypes, including certain age-related diseases. PMID- 12543257 TI - A Darwinian-evolutionary concept of age-related diseases. AB - Humans and animals are structurally designed as a compromise to guarantee optimal survival until the time of reproduction based on natural selection that is effective until that age. Modern conditions of life including improvement of hygiene, preventive and curative medicine as well as socio-economic and political developments have led to an increase of the mean life expectancy that allows ever larger proportions of the population to reach an age that is far beyond that of the reproductive phase. The concept of a Darwinian-evolutionary basis for the development of age-related diseases in principle postulates that genetic traits that are beneficial in younger years to allow for successful reproduction may become deleterious in the elderly, i.e. when selective pressure does not seem to be effective anymore. Examples for this mechanism of pleiotropic antagonism taken from the work of the Institute for Biomedical Aging Research in Innsbruck, Austria, include atherosclerosis, benign and malignant prostate hypertrophy, Alzheimer's disease and the reciprocal relationship between cellular senescence and cancer. PMID- 12543258 TI - Aging, fertility, and immortality. AB - Evolutionary theory suggests that fecundity rates will plateau late in life in the same fashion as mortality rates. We demonstrate that late-life plateaus arise for fecundity in Drosophila melanogaster. The result qualitatively fits the evolutionary theory of late life based on the force of natural selection. But there are a number of alternative interpretations. Fecundity plateaus could be secondary consequences of mortality-rate plateaus. Female fecundity plateaus might arise from diminished male sexual function. Another alternative hypothesis is analogous to male sexual inadequacy: nutritional shortfalls. These may arise later in life because of a decline in female feeding or digestion. If some females have a life-long tendency to lay eggs at a faster rate, but die earlier, then aging for fecundity could arise from the progressive loss of the fast layers, with the late-life plateau simply the laying patterns of individual females who were slow-layers throughout adult life. If this type of model is generally applicable to late life, then we should find that the females who survive to lay at a slow but steady rate in late life have a similar laying pattern in mid-life. PMID- 12543259 TI - Calorie restriction in rhesus monkeys. AB - Calorie restriction (CR) extends lifespan and reduces the incidence and age of onset of age-related disease in several animal models. To determine if this nutritional intervention has similar actions in a long-lived primate species, the National Institute on Aging (NIA) initiated a study in 1987 to investigate the effects of a 30% CR in male and female rhesus macaques (Macaca mulatta) of a broad age range. We have observed physiological effects of CR that parallel rodent studies and may be predictive of an increased lifespan. Specifically, results from the NIA study have demonstrated that CR decreases body weight and fat mass, improves glucoregulatory function, decreases blood pressure and blood lipids, and decreases body temperature. Juvenile males exhibited delayed skeletal and sexual maturation. Adult bone mass was not affected by CR in females nor were several reproductive hormones or menstrual cycling. CR attenuated the age associated decline in both dehydroepiandrosterone (DHEA) and melatonin in males. Although 81% of the monkeys in the study are still alive, preliminary evidence suggests that CR will have beneficial effects on morbidity and mortality. We are now preparing a battery of measures to provide a thorough and relevant analysis of the effectiveness of CR at delaying the onset of age-related disease and maintaining function later into life. PMID- 12543260 TI - Nutritional control of aging. AB - For more than 60 years the only dietary manipulation known to retard aging was caloric restriction, in which a variety of species respond to a reduction in energy intake by demonstrating extended median and maximum life span. More recently, two alternative dietary manipulations have been reported to also extend survival in rodents. Reducing the tryptophan content of the diet extends maximum life span, while lowering the content of sulfhydryl-containing amino acids in the diet by removing cysteine and restricting the concentration of methionine has been shown to extend all parameters of survival, and to maintain blood levels of the important anti-oxidant glutathione. To control for the possible reduction in energy intake in methionine-restricted rats, animals were offered the control diet in the quantity consumed by rats fed the low methionine diet. Such pair-fed animals experienced life span extension, indicating that methionine restriction related life span extension is not a consequence of reduced energy intake. By feeding the methionine restricted diet to a variety of rat strains we determined that lowered methionine in the diet prolonged life in strains that have differing pathological profiles in aging, indicating that this intervention acts by altering the rate of aging, not by correcting some single defect in a single strain. PMID- 12543261 TI - Estrogen, synaptic plasticity and hypothalamic reproductive aging. AB - Unlike primates who undergo ovarian failure and loss of sex steroids at the end of reproduction, aging rodents undergo constant vaginal estrus followed by constant diestrus and finally anestrus, which indicates the absence of responsive ovarian follicles. The latter state is analogous to menopause in women. The timing of the appearance of constant estrus is determined by many factors including estrogen exposure in the brain during development and the number of times that the animal gets pregnant. The chief site of this reproductive aging in rat brains is the arcuate nucleus of the hypothalamus. The transition from normal cycles to constant estrus parallels the females' gradually decreased ability to respond to administered estradiol with a cycle of inhibition followed by disinhibition of gonadotrophin-releasing hormone. Evidence has accumulated indicating this to be due to a loss of the rat's ability to respond to markedly elevated estradiol with the usual arcuate nucleus neuro-glial plasticity that supports the estrogen-induced gonadotrophin surge (EIGS). Just as male rats are not capable of an EIGS, aged females loose this ability through repeated EIGS. Experiments indicate that in male rats the hypothalamic synaptology that develops as a result of exposure to testicular androgens in the perinatal period (brain sexual differentiation) is a result of conversion of testosterone from the testes to estrogen in the brain and is therefore due to early estrogen exposure. Aging females appear to reach a synaptology similar to males and constant estrus as a result of repeated exposure to ovarian estrogens during their reproductive careers. The relative role of aging and hormonal factors remains unclear. Morphological evidence is presented that indicates the above effects of estrogen involve changes in hypothalamic arcuate nucleus neurons and glia, including changes in the organization of perikaryal membranes as well as arcuate nucleus synaptology and the load of peroxidase in the astroglia. A possible role for free radicals (reactive oxygen species) in hypothalamic reproductive aging has been proposed. Such a mechanism is supported by evidence that the anti-oxidant vitamin E delays the onset of constant estrus and the accumulation of glial peroxidase in aging female rats. However, since the synaptology and peroxidase load in constant estrus females is independent of the age at which the constant estrus occurs, it appears that the role of (repeated) estradiol exposure is more deterministic of hypothalamic failure than is aging, per se. PMID- 12543262 TI - The neurobiology of memory changes in normal aging. AB - Cognitive alterations occur over the lifespan of every species studied and have been quantified carefully in humans, other primates and rodents. Correspondingly, changes in hippocampal function have been associated with a number of observed memory impairments across species. It appears that humans, alone, show Alzheimer's disease-like cognitive and neural pathology spontaneously. Thus, a comparison of normal age-related changes in cognition in other animals can help disambiguate the boundary between normal and pathological states of aging in humans. Another important contribution made from studying aging in non-human species is the ability to examine, in more detail, the basic neural mechanisms that may be responsible for brain aging in these species. So far, most of the functional neurobiological studies have been conducted in the aged rat. We propose that the link between rodent and human work can be made much stronger by combining neurophysiological and behavioral investigation of normal aging in the non-human primate. PMID- 12543263 TI - Effects of aging on the hippocampal formation in a naturally occurring animal model of mild cognitive impairment. AB - Vertical integration is being used to great advantage in neurobiological research on the basis of age-related cognitive decline. Such research bridges analysis between the molecular and cellular levels and the outcome of impaired psychological functions. Current use of animals models within this paradigm has defined mild cognitive impairment in a subpopulation of outbred aged Long-Evans rats by assessment of hippocampal-dependent spatial cognition. Aged rats with cognitive impairment exhibited no loss of neurons in the hippocampus. Current research is focused on the functional alterations in neurons by methods which assess transcriptional mechanisms and signaling pathways. PMID- 12543264 TI - Regeneration in the central nervous system. AB - Unlike neonatal axons, mammalian adult axons of the CNS do not regenerate after injury. This developmental loss of regenerative capacity, is correlated with the onset of myelination. Likewise, myelin, or myelin-associated components such as Nogo-A and myelin-associated glycoprotein (MAG) inhibit regeneration from older but not younger neurons. Identification of the molecular events responsible for this developmental loss of regenerative capacity is central to devise strategies to encourage regeneration in adults after injury. Endogenous levels of the cyclic nucleotides cAMP and cGMP have been suggested to determine the neuronal responsiveness to various axonal guidance factors. Elevating cAMP concentrations block Nogo-A or MAG induced inhibition of neurite outgrowth in older neurons, whereas suppressing cAMP levels in young neurons renders them susceptible to Nogo A and MAG. Interestingly, elevated cAMP levels abrogated the Nogo-A and MAG mediated activation of RhoA and down regulation of Rac1 in adult neurons. In contrast, elevation of cAMP leads to the inactivation of RhoA and prevents activation of downstream effector proteins, while Rac is activated. We therefore conclude that the endogenous neuronal cAMP levels determine the neuronal responsiveness to myelin-associated neurite growth inhibitors by regulating rho GTPase activities. PMID- 12543265 TI - Auditory function in presbycusis: peripheral vs. central changes. AB - The hearing abilities of a group of 30 elderly (67-93 yr of age) subjects were compared with those of a group of 30 young (19-27 yr of age) normal hearing volunteers with the aim of characterizing the changes in the peripheral and central parts of the auditory system. In elderly subjects the pure-tone thresholds were typically represented by a gradually sloping curve with a significantly greater decline in men than in women at frequencies of 3 and 4 kHz. In spite of pure tone threshold elevation in the elderly, the difference limen for intensity at 1 and 3 kHz were not significantly smaller than in the young subjects. The incidence and levels of spontaneous, transient and distortion product otoacoustic emissions were low, which would suggest the involvement of outer hair cell pathology. Also, contralateral suppression was less marked in elderly than in young subjects. Speech audiometry in the elderly revealed serious difficulties in understanding speech. Deteriorated temporal resolution, as demonstrated by increased gap detection thresholds, correlated significantly with increased speech recognition thresholds. The results support the view that presbycusis represents a combination of deteriorated function of the auditory periphery with deteriorated function of the central auditory system. PMID- 12543266 TI - Aging and the human neocortex. AB - Neurostereology has been applied to quantitative anatomical study of the human brain. Such studies have included the total neocortical number of neurons and glial cells, the estimated size distribution of neocortical neurons, the total myelinated fiber length in the brain white matter, the total number of synapses in the neocortex, and the effect of normal aging on these structural elements. The difference in total number of neurons was found to be less than 10% over the age range from 20 to 90 years, while the glial cell number in six elderly individuals, mean age 89.2 years, showed an average number of 36 billion glial cells, which was not statistically significantly different from the 39 billion glial cells in the neocortex of six young individuals with a mean age of 26.2 years. The total myelinated fiber length varied from 150,000 to 180,000 km in young individuals and showed a large reduction as a function of age. The total number of synapses in the human neocortex is approximately 0.15 x 10(15) (0.15 quadrillion). Although the effect of aging is seen in all estimated structural elements, the effect of sex is actually higher. The functional relevance of these differences in neuron numbers in both age and gender is not known. PMID- 12543267 TI - Polycyclic phenols, estrogens and neuroprotection: a proposed mitochondrial mechanism. AB - Polycyclic phenols, including the estrogens, have been shown to be potent neuroprotectants in a variety of cellular and animal model systems. Although classical estrogen receptor interactions and consequent responses play a role in certain circumstances, the neuroprotective activity of polycyclic phenols that do not interact with estrogen receptors ERalpha or ERbeta is more likely to be through non-genomic mechanism(s). We propose here that such non-feminizing polycyclic phenols exert their protective effects at least in part by stabilizing mitochondria, preventing apoptotic and/or necrotic forms of cell death that are associated with mitochondrial dysfunction. Consistent with this mitochondrial model and the available data, these compounds protect neurons and other cell types from a wide variety of pathologically relevant stressors. PMID- 12543268 TI - The neuroprotective effects of estrogen on the aging brain. AB - The population of the western world is ageing. This increase in the elderly population will inevitably mean a rise in the prevalence of age-related cognitive decline and late-onset neuropsychiatric disorder, such as Alzheimer's disease (AD). There are sex differences in the incidence and age of onset of these disorders. Sex steroids and sex chromosomes are therefore implicated in their pathophysiology. We have identified relevant past and current literature using a Medline search and from the references of relevant papers. These were then reviewed and relevant articles have been summarized and included in the review. Evidence is presented for the wide-ranging actions of estrogen in the brain at the cellular, metabolic and neurotransmitter levels as well as from the cognitive, AD, depression and cerebrovascular perspectives. The authors conclude that it is unlikely that estrogen will become a stand-alone treatment for any of these disorders, although there may still be a role as an adjunctive treatment and as a prophylactic measure. PMID- 12543269 TI - Cognition enhancing or neuroprotective compounds for the treatment of cognitive disorders: why? when? which? AB - Neurodegenerative disorders such as Alzheimer's disease, Lewy-Body dementia, Parkinson's disease and cerebrovascular dementia result in an insidious cognitive and behavioural decline culminating in the development of severe dementia. Based on current population projections it has been estimated that by 2050 the number of individuals over 65 will increase to 1.1 billion worldwide and as a consequence, the number of cases of dementia to 37 million. Faced with such an enormous public health and socio-economic burden it is evident that the importance of therapeutic intervention aimed at either finding a cure or preventing disease progression cannot be overstated. The aim of the present paper is to present an overview, in the context of a brain aging continuum, at what stage cognition enhancing and/or neuroprotective intervention strategies aimed at stabilising and/or preventing neurodegenerative disease could demonstrate potential clinical benefit. In particular, the clinical identification of patients with mild cognitive impairment and age-associated memory impairment which may represent a 'transition' state between normal aging and dementia is discussed as a potential clinical population cohort targeted for early intervention in dementia. Considering the wide spectrum of cognitive and psychotic effects in dementia juxtaposed with the neuropathological evolution of the disease, it is clear that a variety of therapeutic intervention(s) will be required in order, to at the least, stabilise disease progression. Evidently, since Alzheimer's disease is by far the most prevalent form of dementia, and will undoubtedly serve as the benchmark for any future treatment of dementia, an update of current symptomatic and disease-modifying therapeutic approaches (cholinergic, glutamatergic, nootropics, beta-amyloid cascade inhibitors) will be reviewed. PMID- 12543270 TI - Body size, insulin/IGF signaling and aging in the nematode Caenorhabditis elegans. AB - A number of recent studies of aging in Drosophila, mice and dogs have shown an association between reduced body size and increased lifespan. It is unclear (a) whether such an association is a general feature of animal species; and (b) whether the association reflects an effect of body size on aging, or pleiotropic effects of common determinants of growth and aging. To address these issues, we have studied the relationship between size and lifespan in the nematode Caenorhabditis elegans, and surveyed related findings in Drosophila. In C. elegans, we compared 12 wild isolates with varying body size and lifespan, but saw no correspondence between these traits. We also examined aging in giant and dwarf mutants, but observed only reduced lifespan in all cases. In a comparison of 15 long-lived daf-2 insulin/IGF receptor mutants, we saw a positive correlation between body length and lifespan, and up to a 28% increase in daf-2 mutant body volume. Thus, in C. elegans, insulin/IGF signaling may limit growth rather than promote it. Studies of Drosophila show no consistent correlation between body size and lifespan. These results indicate that the negative correlation between body size and lifespan seen in some mammals is not typical of invertebrates, but support the view that co-variation of size and longevity may occur via effects on insulin/IGF signaling. PMID- 12543271 TI - Glucose regulation and oxidative stress in healthy centenarians. AB - Aging, oxidative stress and insulin resistance are strongly correlated. There is a growing body of evidence showing that aging is associated with a significant rise in oxidative stress mainly due to a decline in anti-oxidant activity and a rise in pro-oxidant factors such as glucose and insulin concentrations. Furthermore, aging is also associated with a progressive rise in insulin resistance which is due to a complex network of environmental, anthropometric and neuro-hormonal factors. It is noteworthy that extreme longevity, e.g. centenarians, is associated with a low degree of oxidative stress and insulin resistance. The causes for such differences between aged subjects and centenarians is not fully understood. It is likely that a specific genetic background might play a role. However, the insulin gene does not seem to be involved for explaining such age-related differences. PMID- 12543272 TI - The pharmacology of amyloid precursor protein processing. AB - The possibility to understand the causes and treat the symptoms of Alzheimer's disease patients is still a great challenge. The triggering events leading to the selective neurodegeneration observed in Alzheimer's brains are not completely understood. This lack of understanding of the pathophysiological processes posses an important theoretical challenge for the rational design of pharmacological intervention. The scientific community is divided over the pathogenesis of the disease which is historically divided between 'baptists' and 'tauists'. Baptists suggest that beta-amyloid, the peptide deposited in neuritic plaques, is the cause of all damages while tauists suggest that hyperphosphorylated tau, the cytoskeletal protein that forms neurofibrillary tangles, is the culprit for the disease. This review will be focused on the pharmacological modulation of the amyloid precursor protein metabolism, with the goal of reducing the formation of beta-amyloid. Over the years such an approach has led to the identification of a complex intracellular mechanism, which may be regulated by neurotransmitters and other ligands. More recently, these efforts have contributed to the characterization of the enzymes which regulate the formation of beta-amyloid. PMID- 12543273 TI - The serotonin 5-HT4 receptor and the amyloid precursor protein processing. AB - A large body of evidence supports a major role for the serotonin 5-HT(4) receptor in learning and memory and it is suggested that 5-HT(4) agonists may be beneficial for memory disorders such as Alzheimer's disease (AD). The 5-HT(4) receptors are members of the G protein-coupled receptor superfamily and are positively coupled to adenylyl cyclase. In this communication we show that a neuronal isoform of the human 5-HT(4) receptor, h5-HT(4(g)) regulates the metabolism of the amyloid precursor protein (APP695). This process is observed in Chinese hamster ovary (CHO) cells stably coexpressing the neuronal h5-HT(4(g)) receptor isoform as well as the human APP695. The 5-HT(4) agonists strongly stimulate the release of the non-amyloidogenic soluble amyloid precursor protein sAPPalpha as detected by immunoblot. Prucalopride was more potent than serotonin (5-HT) with regard to enhanced of sAPPalpha secretion. This process was blocked by a selective 5-HT(4) antagonist, GR113808. Furthermore, 5-HT(4) ligands enhance sAPPalpha secretion via cAMP-dependent and PKA-independent signalling pathways indicating there are alternative pathways by which the h5-HT(4) receptor via cAMP regulates APP metabolism. Because the alpha-cleavage event may preclude the formation of amyloidogenic peptides, and secreted sAPPalpha has putative neuroprotective and enhancing-memory properties, our present data suggest the 5 HT(4) receptor as a novel target for the treatment of AD. PMID- 12543274 TI - Post-mortem brain tissue cultures from elderly control subjects and patients with a neurodegenerative disease. AB - Aging may be viewed as a progressive loss of normal biological function. Due to complex genetic and environmental interactions, the sequence of functional impairment shows a high degree of individual variability. In humans life style and health care have an additional influence on the aging process. To study aging and age-related disorders of the human nervous system, brain tissue that has undergone aging and pathological alterations can provide valuable study material. Recently, we have shown that adult human postmortem brain tissue can be cultured and experimentally manipulated. This approach permits the study of cellular aspects of human neuronal aging and neurodegenerative processes and complements those existing research methods such as in vivo imaging (MRI, PET, etc.) and fixed or frozen postmortem brain tissue examination. PMID- 12543275 TI - Molecular manipulation of neural precursors in situ: induction of adult cortical neurogenesis. AB - Over the past three decades, research exploring potential neuronal replacement therapies have focused on replacing lost neurons by transplanting cells or grafting tissue into diseased regions of the brain. Over most of the past century of modern neuroscience, it was thought that the adult brain was completely incapable of generating new neurons. However, in the last decade, the development of new techniques has resulted in an explosion of new research showing that neurogenesis, the birth of new neurons, normally occurs in two limited and specific regions of the adult mammalian brain, and that there are significant numbers of multipotent neural precursors in many parts of the adult mammalian brain. Recent findings from our lab demonstrate that it is possible to induce neurogenesis de novo in the adult mammalian brain, particularly in the neocortex where it does not normally occur, and that it may become possible to manipulate endogenous multipotent precursors in situ to replace lost or damaged neurons. Recruitment of new neurons can be induced in a region-specific, layer-specific, and neuronal type-specific manner, and newly recruited neurons can form long distance connections to appropriate targets. Elucidation of the relevant molecular controls may both allow control over transplanted precursor cells and potentially allow the development of neuronal replacement therapies for neurodegenerative disease and other CNS injuries that do not require transplantation of exogenous cells. PMID- 12543276 TI - Brain ischemia as a potential target of gene therapy. AB - Brain infarction is one of the most important age-associated medical conditions, and the age-related neuronal vulnerability to brain ischemia is suggested to play an important role. Recent advancements in gene transfer techniques have provided promising approaches to the treatment of brain ischemia. In experimental studies, the ischemic penumbra area can be targeted by gene transfer even after ischemic insult, and post-ischemic gene therapy seems effective in attenuation of ischemic damage in both global and focal brain ischemia. Perivascular approaches of gene transfer to the cerebral blood vessels through the subarachnoid space may lead to prevention of brain ischemia caused by vasospasm after subarachnoid hemorrhage. Gene transfer to cerebral blood vessels and ischemic brain tissue may offer future therapeutic approaches to stroke. PMID- 12543277 TI - Selected aspects of endocrine pharmacology of the aging male. AB - This minireview explores the endocrinology and the clinical consequences of age related hypogonadism (hypotestosteronemia). In addition, pharmacological and clinical applicability of new androgen formulations is described briefly. Other topics include selective androgen receptor modulators, non-feminizing estrogens, and the possible use of selective aromatase modulators. Finally, a theoretical concept of hormone displacement (i.e. excessive hormone production) is introduced using cortisol as an example. PMID- 12543278 TI - Melatonin rhythmicity: effect of age and Alzheimer's disease. AB - The circadian rhythm of the pineal gland hormone, melatonin is generated within the hypothalamic suprachiasmatic nuclei (SCN), site of the circadian clock. The circadian clock and its output melatonin rhythm is synchronized to the 24h day by environmental light which is transmitted from the retina to the SCN primarily via the retinohypothalamic tract. Changes in both the amplitude and timing of the melatonin rhythm have been reported with aging in humans. Whether these age related changes (reduced melatonin amplitude, earlier timing of melatonin rhythm) are a result of aging of the retina, the SCN clock, the pineal gland, their neural connections or a combination of some or all of these is not known. The fragmented sleep/wake patterns observed in the elderly and to a greater extent in patients with Alzheimer's disease have been shown to be partly related to an altered retina-SCN-pineal axis. Therapies designed to reinforce the circadian axis (for example, administration of melatonin or light) have been reported to alleviate the disturbed circadian rhythms and disrupted sleep. Future research needs to pinpoint the site(s) of age-related dysfunction so that therapies can be specifically tailored to correct the abnormality in addition to reinforcing any of the intact processes. PMID- 12543279 TI - Dawn-dusk simulation light therapy of disturbed circadian rest-activity cycles in demented elderly. AB - We investigated whether low intensity dawn-dusk simulation (DDS), a 'naturalistic' form of light therapy designed to embed sleep in its accustomed phase, could improve the disturbed circadian rest-activity cycle, nocturnal sleep and and/or cognitive functions in dementia. A protocol of 3 weeks each of baseline, treatment and follow-up was completed by 13 patients (85yr old+/-5yr, MMSE 14+/-5; n=9 DDS versus n=4 'placebo' dim red light) who wore an activity/lux monitor throughout. There were no significant changes in clinical or cognitive status, nor modification of circadian stability or amplitude characteristics of the rest-activity cycle. However, two aspects of sleep responded to DDS but not to dim red light. The main sleep episode was 1:14h earlier during treatment (p=0.03) compared with before and after DDS. With respect to actimetry-determined sleep variables, the DDS group tended to have shortened 'sleep latency', longer 'sleep duration', more nocturnal immobility and less nocturnal activity than the dim red group (p<0.1). In parallel, nighttime light exposure tended to be reduced (p=0.07). These promising findings-after only 3 weeks of light treatment in elderly patients with advanced dementia-suggest that the circadian timing system remains functionally responsive even to low intensity DDS light. Increasing zeitgeber strength is an important strategy for improving sleep quality and timing in dementia, and DDS light therapy may provide one of the appropriate means to do so. PMID- 12543281 TI - Clinical effectiveness of laparoscopic fundoplication in a U.S. community. AB - BACKGROUND: The aim of our study was to determine the outcome of laparoscopic fundoplication for reflux disease in a cohort of patients who underwent this procedure in routine clinical practice. METHODS: We identified 151 patients who had undergone laparoscopic fundoplication in a managed care organization in Milwaukee. Symptoms were evaluated using a validated questionnaire. Postoperative medication use and endoscopic and surgical procedures were recorded. RESULTS: Eighty-seven patients agreed to participate, of whom 80 (41 [51%] men) were eligible. Their mean (+/- SD) age was 45 +/- 12 years, and the mean duration after surgery was 20 +/- 10 months. Thirty-six patients (45%) underwent the procedure because their physician recommended it, and 22 (27%) because they thought it would cure their disease. Forty-three patients (61%) were satisfied with the outcome of the procedure. Twenty-six patients (32%) were taking medications on a regular basis for treatment of heartburn, 9 (11%) required esophageal dilation for dysphagia, and 6 (7%) had repeat surgical procedures. Of the 54 patients (67%) who reported new symptoms after surgery, 38 reported excessive gas, 22 reported abdominal bloating, and 22 reported dysphagia. Health related quality of life was significantly lower in patients with these symptoms. CONCLUSION: Medical therapy is required for control of heartburn in approximately one third of patients after laparoscopic fundoplication. New symptoms are common after surgery. Patients need to be better informed about the indications and outcomes of surgery. PMID- 12543282 TI - Bowel dysfunction after laparoscopic antireflux surgery: incidence, severity, and clinical course. AB - PURPOSE: To evaluate the incidence, severity, and clinical course of postoperative bowel dysfunction, primarily diarrhea, after laparoscopic antireflux surgery. METHODS: Patients who underwent laparoscopic antireflux surgery during January to December 1998 responded to a questionnaire about pre existing and postoperative bowel symptoms, which included questions about the type of bowel dysfunction (diarrhea, abdominal pain, bloating, constipation), onset in relation to surgery, frequency, severity, duration, use of medical resources or diagnostic evaluations, and treatment outcome. RESULTS: Of the 109 patients who underwent laparoscopic antireflux surgery at our center during the study, 84 (77%) completed the survey. Thirty-six (43%) had no bowel dysfunction before or after surgery, whereas 29 (35%) had pre-existing bowel dysfunction. New bowel symptoms developed postoperatively in 30 patients (36%), including bloating in 16 (19%) and diarrhea in 15 (18%). Two thirds of the patients with new diarrhea developed it within 6 weeks after surgery. The severity of the diarrhea ranged from mild to debilitating; 4 had fecal incontinence. Most patients (13/15) with diarrhea had symptoms for > or =2 years following surgery. No patient was hospitalized, and only 2 patients reported temporary work loss. CONCLUSION: Postoperative bowel dysfunction, namely diarrhea, is an important adverse effect of antireflux surgery. Awareness of this complication should lead to prompt recognition, effective management, and reduction in anxiety. PMID- 12543283 TI - The risk of osteoporosis in Caucasian men and women with obstructive airways disease. AB - PURPOSE: Because patients with obstructive airways disease may be susceptible to osteoporosis, we sought to determine the association between airflow obstruction and osteoporosis. SUBJECTS AND METHODS: We analyzed data from Caucasian participants (n = 9502) in the Third National Health and Nutrition Examination Survey, conducted in the United States between 1988 and 1994. We used data from dual-energy x-ray absorptiometry measurements of the total femur to determine whether a study participant had osteoporosis (defined as total bone mineral density values < or =2.5 SD below the corresponding mean values from young, healthy participants). We calculated the odds ratio (OR) for osteoporosis in four lung function categories: none, mild, moderate, and severe airflow obstruction. RESULTS: Overall, airflow obstruction was associated with increased odds of osteoporosis compared with without airflow obstruction (OR = 1.9; 95% confidence interval [CI]: 1.4 to 2.5). Participants with severe airflow obstruction were at especially increased risk (OR = 2.4; 95% CI: 1.3 to 4.4). Moderate but not mild airflow obstruction was also associated with osteoporosis. CONCLUSION: Airflow obstruction was an important risk factor for osteoporosis in the study population. These data highlight the importance of measuring bone mineral density in those with moderate-to-severe airflow obstruction for the detection and prevention of osteoporosis-related morbidity. PMID- 12543284 TI - Early antidepressant therapy for elderly patients. AB - PURPOSE: We studied factors affecting the management of depression in older patients, especially the use of early antidepressant therapy. METHODS: We recruited 128 primary care physicians to view one version of a 5-minute videotape of an elderly patient with somatic symptoms that were suggestive of depression, and to complete an interview that assessed decision making. Using an experimental factorial design, 16 versions of the videotape were produced, holding constant the clinical features of the case, while varying the patient's age, race, sex, and socioeconomic status. Dependent variables were the physicians' probability assessment of depression and the recommendation of antidepressant medication after the first visit. RESULTS: Depression was considered a possible diagnosis by 121 physicians (95%) and the most likely diagnosis by 69 (54%). Sixteen physicians (13%) recommended antidepressant therapy after the first visit, and they were less likely than other physicians to order initial laboratory tests to assess the possibility of other conditions. Recommendations for antidepressant therapy was not associated with patient age, sex, race, or socioeconomic status, or with physician sex, race, or experience. Family physicians were more likely than internists to recommend an antidepressant (19% [12/64] vs. 6% [4/64], P = 0.04). CONCLUSION: Based on a 5-minute vignette, physicians were likely to recognize depression, independent of patient characteristics. Those recommending early antidepressant therapy were more likely to be in family medicine and less likely to investigate other diagnoses initially. PMID- 12543285 TI - Valvular flow abnormalities are often identified by a resting focused Doppler examination performed at the time of stress echocardiography. AB - BACKGROUND: Patients are usually referred for stress echocardiography to assess whether there is inducible myocardial ischemia. At some centers, a focused Doppler examination is also performed. We sought to determine the clinical value of this additional study by examining how often valvular flow abnormalities were identified that might affect clinical care. METHODS: We reviewed 1272 consecutive stress echocardiogram reports from a 1-year period, including 1223 tests that contained focused Doppler data. Important Doppler findings were defined as at least moderate mitral regurgitation, at least mild aortic regurgitation, any aortic or mitral valve stenosis, or any resting left ventricular outflow tract gradient. RESULTS: Overall, focused Doppler identified an important Doppler abnormality in 214 patients (17%). At least moderate mitral regurgitation was identified in 67 patients (5%) and at least mild aortic regurgitation was identified in 163 patients (13%). In addition, aortic stenosis (n = 14; 1%), mitral stenosis (n = 5; 0.4%), and resting outflow tract gradient (n = 2; 0.2%) were noted. A prior echocardiogram had been performed at our institution in 317 patients (26%). For this subset, a new important Doppler finding, a two-step change in regurgitant grade, or a one-step change in stenosis severity was noted in 28 patients (9%). Among patients who had a previous study, the prevalence of new findings was the same (9%) in those who had been studied within the previous year as in those whose previous study had been performed more than 2 years before. CONCLUSION: Focused Doppler documents valvular flow abnormalities in 17% of patients referred for stress echocardiography, thereby enhancing the potential overall value of the test. The frequency of new findings was independent of the time interval from the previous Doppler study. These data should be considered when clinicians choose a stress imaging modality. PMID- 12543286 TI - Association between plasma chromogranin A concentration and long-term mortality after myocardial infarction. AB - PURPOSE: Chromogranin A, a polypeptide that is distributed throughout the neuroendocrine system, may be a marker of neuroendocrine activation. We sought to assess the long-term prognostic value of circulating levels of chromogranin A after myocardial infarction. METHODS: We studied 119 patients (88 [74%] male; median age, 70 years [interquartile range, 62 to 75 years]) with documented myocardial infarction. Chromogranin A levels in plasma were determined by radioimmunoassay from samples obtained 3 days after the onset of symptoms. RESULTS: During a median follow-up of 10.8 years, 56 patients (47%) died. The median concentration of chromogranin A in plasma was 24 ng/mL (interquartile range, 18 to 36 ng/mL). Plasma chromogranin A levels were associated with increased long-term mortality (hazard ratio [HR] = 1.17 per 10-ng/mL increase; 95% confidence interval [CI]: 1.06 to 1.28) in models that adjusted for age, clinical heart failure during the initial hospitalization, and use of thrombolytic therapy. As a dichotomous variable (cutoff, 24 ng/mL), an elevated chromogranin A level was also associated with mortality in univariate analysis (HR = 2.6; 95% CI: 1.4 to 4.8), but this relation was no longer significant after adjustment for age (HR = 1.4; 95% CI: 0.8 to 2.7). CONCLUSION: Plasma levels of chromogranin A are related to long-term mortality after myocardial infarction, perhaps because they reflect neuroendocrine activation. PMID- 12543287 TI - What affects influenza vaccination rates among older patients? An analysis from inner-city, suburban, rural, and Veterans Affairs practices. AB - BACKGROUND: Despite strong evidence of the effectiveness of influenza vaccination, immunization rates have reached a plateau that is below the 2010 national goals. Our objective was to identify facilitators of, and barriers to, vaccination in diverse groups of older patients. METHODS: A survey was conducted in 2000 by computer-assisted telephone interviewing of patients from inner-city health centers, Veterans Affairs (VA) outpatient clinics, rural practices, and suburban practices. The inclusion criteria were age > or =66 years and an office visit after September 30, 1998. RESULTS: Overall, 1007 (73%) interviews were completed among 1383 patients. Influenza vaccination rates were 91% at VA clinics, 79% at rural practices, 79% at suburban practices, and 67% at inner-city health centers. There was substantial variability in vaccination rates among practices, except at the VA. Nearly all persons who were vaccinated reported that their physicians recommended influenza vaccinations, compared with 63% of unvaccinated patients (P <0.001). Thirty-eight percent of unvaccinated patients were concerned that they would get influenza from the vaccine, compared with only 6% of vaccinated persons (P <0.001). Sixty-three percent of those vaccinated, in contrast with 22% of unvaccinated persons, thought that an unvaccinated person would probably contract influenza (P <0.001). CONCLUSION: Older patients need intentional messages from physicians that recommend vaccination. Furthermore, more patient education is needed to counter myths about adverse reactions. PMID- 12543288 TI - Medication nonadherence and the outcomes of patients with quiescent ulcerative colitis. AB - PURPOSE: We conducted a prospective study to determine the effects of nonadherence with mesalamine among patients with quiescent ulcerative colitis. METHODS: We followed a cohort of 99 consecutive patients who had ulcerative colitis in remission for more than 6 months and who were taking maintenance mesalamine. Medication adherence rates were calculated based on pharmacy records and a validated formula. Nonadherence was defined as refilling less than 80% of prescribed medication. Patients were followed prospectively and evaluated either in clinic or via telephone at 6, 12, and 24 months. The primary outcome was clinical recurrence of ulcerative colitis. Proportional hazards models were used to adjust for confounders. RESULTS: At 6 months, 12 patients (12%) had clinical recurrence of disease symptoms, all of whom were nonadherent with medication. At 12 months, 19 of 86 patients had recurrent disease, 13 (68%) of whom were nonadherent. Patients who were not adherent with medication had more than a fivefold greater risk of recurrence than adherent patients (hazard ratio = 5.5; 95% confidence interval: 2.3 to 13; P < 0.001). CONCLUSION: Nonadherence with medication increases the risk of clinical relapse among patients with quiescent ulcerative colitis. Future research should be directed at behavioral interventions to improve adherence. PMID- 12543289 TI - Tachycardia-induced cardiomyopathy. AB - Systolic dysfunction associated with chronic tachyarrhythmias, known as tachycardia-induced cardiomyopathy, is a reversible form of heart failure characterized by left ventricular dilatation that is usually reversible once the tachyarrhythmia is controlled. Its development is related to both atrial and ventricular arrhythmias. The diagnosis is usually made following observation of a marked improvement in systolic function after normalization of heart rate. Clinicians should be aware that patients with unexplained systolic dysfunction may have tachycardia-induced cardiomyopathy, and that controlling the arrhythmia may result in improvement and even complete normalization of systolic function. PMID- 12543290 TI - Use of random skin biopsy to diagnose intravascular lymphoma presenting as fever of unknown origin. PMID- 12543291 TI - Systemic symptoms without erythema migrans as the presenting picture of early Lyme disease. PMID- 12543292 TI - Efficacy of molecular adsorbent recirculating system for the treatment of intractable pruritus in cholestasis. PMID- 12543293 TI - Failure of health care professionals to interpret fecal occult blood tests accurately. PMID- 12543294 TI - Let the patient beware: the evolving truth about laparoscopic antireflux surgery. PMID- 12543295 TI - Toward a more complete appreciation of the clinical spectrum of Borrelia burgdorferi infection: early Lyme disease without erythema migrans. PMID- 12543296 TI - The importance of using strict inclusion criteria in subclinical hypothyroid studies. PMID- 12543297 TI - Methodological problems in a study on the treatment of subclinical hypothyroidism. PMID- 12543298 TI - Clopidogrel-associated angioedema. PMID- 12543299 TI - Caloric intake, not carbohydrate or fat consumption, determines weight loss. PMID- 12543373 TI - Critical review of acylation-stimulating protein physiology in humans and rodents. AB - In the last few years, there has been increasing interest in the physiological role of acylation-stimulating protein (ASP). Recent studies in rats and mice, in particular in C3 (-/-) mice that are ASP deficient, have advanced our understanding of the role of ASP. Of note, the background strain of the mice influences the phenotype of delayed postprandial triglyceride clearance in ASP deficient mice. Administration of ASP in all types of lean and obese mice studied to date, however, enhances postprandial triglyceride clearance. On the other hand, regardless of the background strain, ASP-deficient mice demonstrate reduced body weight, reduced leptin and reduced adipose tissue mass, suggesting that ASP deficiency results in protection against development of obesity. In humans, a number of studies have examined the relationship between ASP, obesity, diabetes and dyslipidemia as well as the influence of diet, exercise and pharmacological therapy. While many of these studies have small subject numbers, interesting observations may help us to better understand the parameters that may influence ASP production and ASP action. The aim of the present review is to provide a comprehensive overview of the recent literature on ASP, with particular emphasis on those studies carried out in rodents and humans. PMID- 12543374 TI - Hydration pressure and phase transitions of phospholipids. I. Piezotropic approach. AB - Dehydration reduces the main phase transition pressure of phospholipids. An analysis based on the Gibbs-Duhem equation shows how the shift of the transition pressure is correlated to the hydration pressure. By using Fourier transform infrared (FT-IR) spectroscopy we determined the hydration-dependent phase transition pressure. The application of our new approach gives hydration pressure values which agree with the values obtained with the osmotic stress method. PMID- 12543375 TI - Hydration pressure and phase transitions of phospholipids. II. Thermotropic approach. AB - It is widely known that dehydration increases the main phase transition temperature of phospholipids. A mathematical analysis now shows that hydration pressure can be calculated by the dehydration-induced shift of the phase transition temperature. The hydration-dependent piezotropic and thermotropic phase transitions were determined by using calorimetry and FT-IR spectroscopy, and the application of our approach gives hydration pressure parameters that agree very well with the values obtained with the osmotic stress method. PMID- 12543376 TI - A permeability transition in liposomes induced by the formation of Ca2+/palmitic acid complexes. AB - Formation of palmitic acid/Ca(2+) (PA/Ca(2+)) complexes was suggested to play a key role in the non-classical permeability transition in mitochondria (NCPT), which seems to be involved in the PA-induced apoptosis of cardiomyocytes. Our previous studies of complexation of free fatty acids (FFA) with Ca(2+) showed that long-chain (C:16-C:22) saturated FFA had an affinity to Ca(2+), which was much higher than that of other FFA and lipids. The formation of FFA/Ca(2+) complexes in the black-lipid membrane (BLM) was demonstrated to induce a nonspecific ion permeability of the membrane. In the present work, we have found that binding of Ca(2+) to PA incorporated into the membrane of sulforhodamine B (SRB)-loaded liposomes results in an instant release of a part of SRB, with the quantity of SRB released depending on the concentration of PA and Ca(2+). The pH optimum of this phenomenon, similar to that of PA/Ca(2+) complexation, is in the alkaline range. The same picture of SRB release has been revealed for stearic, but not for linoleic acid. Along with Ca(2+), some other bivalent cations (Ba(2+), Sr(2+), Mn(2+), Ni(2+), Co(2+)) also induce SRB release upon binding to PA-containing liposomes, while Mg(2+) turns out to be relatively ineffective. As revealed by fluorescence correlation spectroscopy, the apparent size of liposomes does not alter after the addition of PA, Ca(2+) or their combination. So it has been supposed that the cause of SRB release from liposomes is the formation of lipid pores. The effect of FFA/Ca(2+)-induced permeabilization of liposomal membranes has several analogies with NCPT, suggesting that both these phenomena are of similar nature. PMID- 12543377 TI - No entry for TAT(44-57) into liposomes and intact MDCK cells: novel approach to study membrane permeation of cell-penetrating peptides. AB - Cell penetrating peptides (CPPs) have been postulated to carry macromolecules across cell plasma membranes without the need of receptors, transporters, endocytosis or any energy-consuming mechanism. We developed an assay to study lipid bilayer permeation of CPPs. HIV-1 TAT peptides were conjugated to N-(4 carboxy-3-hydroxyphenyl)maleimide (SAM) and incubated with Tb(3+)-containing liposomes. Upon chelation of Tb(3+) by an aromatic carboxylic acid, the fluorescence of Tb(3+) increases many fold. The CPP TAT(44-57)-SAM and TAT(37-53) SAM, as a negative control, were unable to enter liposomes consisting of phosphatidylcholine (PC) or a mix of PC, negatively charged lipids and cholesterol. In parallel, cell entry of fluorescein-labeled TAT peptides was studied using confocal laser scanning microscopy (CLSM). TAT(44-57)-fluorescein did not enter Madin Darby canine kidney (MDCK) cells with intact plasma membranes but accumulated at their basal side. Only cells with impaired plasma membranes, as identified by nuclear staining with ethidium homodimer-1 (EthD-1), showed accumulation of TAT(44-57). Our findings change the perspectives of the potential use of TAT peptides as carriers for intracellular targeting. SAM- and fluorescein labeled TAT(44-57) cannot penetrate lipid bilayers and intact plasma membranes of MDCK cells, respectively. PMID- 12543378 TI - Sensitivity of oocyte-expressed epithelial Na+ channel to glibenclamide. AB - The effect of glibenclamide on heterologously expressed amiloride-sensitive sodium channels (ENaCs) was investigated in Xenopus oocytes. The ENaC is a heteromer and consists of alpha-, beta- and gamma-subunits and the alpha- and beta-subunits have previously been shown to confer sensitivity to glibenclamide. We coexpressed either colonic rat alpha- (ralpha) or guinea-pig alpha-subunit (gpalpha) with Xenopus betagamma-subunits. The gpalphaxbetagamma was significantly stimulated by glibenclamide (100 microM) (184+/-15%), whereas the ralpha-combination was slightly down-regulated by the sulfonylurea (79+/-4%). The stimulating effect did not interfere with Na(+)-self-inhibition resulting from intracellular accumulation of Na(+)-ions. We exchanged cytosolic termini between both orthologs but the gpalpha-chimera with the termini from rat retained sensitivity to glibenclamide. The effect of glibenclamide on Xenopus ENaC (xENaC) was inhibited by ADP-beta-S but not by ATP-gamma-S, when applied intracellularly. Intracellular loading with Na(+)-ions after inhibition of Na(+)/K(+)-ATPases with ouabain prevented an up-regulation of ENaC activity by glibenclamide. Pretreatment of oocytes expressing xENaC with edelfosine (ET-18-OCH(3)) slightly reduced stimulation of I(ami) (118+/-12%; control: 132+/-9%) while phosphatidylinositol-4,5-biphosphate (PIP(2)) significantly reduced the effect of glibenclamide to 101+/-3%. PMID- 12543379 TI - Low chemical specificity of the nicotinic acetylcholine receptor sterol activation site. AB - The nicotinic acetylcholine receptor (nAcChoR) has an absolute requirement for cholesterol if agonist-stimulated channel opening is to occur [Biochemistry 25 (1986) 830]. Certain non-polar analogs could replace cholesterol in vectorial vesicle permeability assays. Using a stopped-flow fluorescence assay to avoid the limitations of permeability assays imposed by vesicle morphology, it was shown that polar conjugates of cholesterol could also satisfy the sterol requirement [Biochim. Biophys. Acta 1370 (1998) 299]. Here this assay is used to explore the chemical specificity of sterols. Affinity-purified nAcChoRs from Torpedo were reconstituted into bilayers at mole ratios of 58:12:30 [1,2-dioleoyl-sn-glycero-3 phosphocholine (DOPC)/1,2-dioleoyl-sn-glycero-3-phosphate (DOPA)/steroid]. When the enantiomer of cholesterol was used, or when the stereochemistry at the 3 hydroxy group was changed from beta to alpha by substituting epicholesterol for cholesterol, activation was still supported. The importance of cholesterol's planar ring structure was tested by comparing planar cholestanol (5alpha cholestan-3beta-ol) with nonplanar coprostanol (5beta-cholestan-3beta-ol). Both supported activation. Thus, these steroids support activation independent of structural features known to be important for modulation of lipid bilayer properties. This provides indirect support for a steroid binding site possessing very lax structural requirements. PMID- 12543380 TI - Protons and calcium alter gating of the hyperpolarization-activated cation current (I(h)) in rod photoreceptors. AB - We investigated the effects of protons and calcium ions on the voltage-dependent gating of the hyperpolarization-activated, nonselective cation channel current, I(h), in rod photoreceptors. I(h) is a cesium-sensitive current responsible for the peak-plateau sag during the rod response to bright light. The voltage dependence of I(h) activation shifted about 5 mV per pH unit, with external acidification producing positive shifts and alkalinization producing negative shifts. Increasing external [Ca(2+)] from 3 to 20 mM resulted in a large (approximately 17 mV) positive shift in I(h) activation. External [Ca(2+)] (20 mM) blocked pH-induced shifts in activation. Cytoplasmic acidification produced by 25 mM sodium acetate led to a negative shift in inactivation (-9 mV) and internal alkalinization produced with 20 mM ammonium chloride resulted in a positive shift (+6 mV). Surface charge binding and screening theory (Gouy-Chapman Stern) accounted for the observed shifts in I(h) activation, with the best fit achieved when protons and calcium ions were assumed to bind to distinct sites on the membrane. Since light induces changes in the retinal ionic environment, these results permit us to gauge the degree to which rod light responses could be modified via alterations in I(h) activation. PMID- 12543381 TI - Thermotropic phase behaviour of alpha-dipalmitoylphosphatidylcholine multibilayers is influenced to various extents by carotenoids containing different structural features--evidence from differential scanning calorimetry. AB - Carotenoids are the effective modulators of physical properties of model and natural membranes. To demonstrate the relationship between the structure of carotenoids and their effect on the molecular dynamics of membranes, we have investigated the influence of five structurally different carotenoids: beta carotene, lycopene, lutein, violaxanthin, zeaxanthin and additionally carotane--a fully saturated derivative of beta-carotene, on thermotropic phase behaviour of dipalmitoylphosphatidylcholine (DPPC) multilamellar vesicles by means of differential scanning calorimetry (DSC). The results obtained indicate that the carotenoids used modulated the thermotropic properties of multibilayers to various extents, broadening the pretransition and the main phase transition peaks and shifting them to lower temperatures. Pronounced decrease of pretransition enthalpy (DeltaH(p)) proves that carotenoids very strongly alter the membrane properties in its gel phase. Comparison of the influence of several carotenoids shows that a rigid, polyisoprenoid chain plays a basic role in altering the thermotropic properties of such membranes and the presence of rings without oxygen-containing groups has a minor significance for the observed interactions. Carotenoids containing epoxy and/or hydroxy groups attached to their rings modify the thermotropic phase behaviour of DPPC multilamellar vesicles stronger than carotenes--a result of their orientation in the DPPC bilayer. PMID- 12543382 TI - Beauvericin-induced channels in ventricular myocytes and liposomes. AB - The antibiotic Beauvericin (BEA) was previously shown to express ionophoric properties under simple experimental systems. Its channel-forming activity was examined in inside-out patches of ventricular myocytes and synthetic membranes with the patch clamp and fluorescence imaging techniques. Current transitions to several open state levels were evident after wash-in. The BEA channel is cation selective. Conductance and kinetics are presented for K(+) and Na(+) substates and main states. The pore was blocked by La(3+). In myocytes, the [K(+)](i) was reduced, while [Na(+)](i) and [Ca(2+)](i) increased, leading to cytolysis. These results indicate that BEA forms cation-selective channels in lipid membranes, which can affect the ionic homeostasis. PMID- 12543383 TI - Design and characterization of enzymosomes with surface-exposed superoxide dismutase. AB - Superoxide dismutase (SOD) was chemically modified by covalent linkage of fatty acid chains to the accessible epsilon-amino groups of the enzyme. This acylation method gave rise to a different enzyme entity (Ac-SOD) as evidenced by different physicochemical properties such as octanol/water partition coefficient and isoelectric point (pI) as compared to SOD. Ac-SOD was incorporated in conventional and long-circulating liposomes (LCL) and characterized in terms of incorporation efficiency, protein to lipid ratio (Prot/Lip), enzymatic activity retention and zeta potential. The observation that Ac-SOD liposomes present enzymatic activity on their external surface indicates that these formulations can act independent of rate and extent of enzyme release as required in case of SOD liposomes. The decrease of superficial charge of liposomal formulations containing Ac-SOD, as compared to SOD liposomes, may be related to the negatively charged enzyme molecules localized on the liposome surface. The comparative characterization of Ac-SOD and SOD liposomal formulations evidenced that the two enzyme forms differ substantially regarding their intraliposomal location: SOD tends to be localized in the internal aqueous spaces, whereas Ac-SOD is expected to be localized in the lipid bilayers of the liposomes, partially buried into the outer surface and exposed to the external medium. These liposomal structures with surface-exposed SOD were designated as Ac-SOD enzymosomes. The properties of these enzymosomes may influence the therapeutic effect, as the release of the enzyme from extravasated vesicles is no longer a necessary requirement for achieving dismutating activity within the inflamed target site. PMID- 12543384 TI - Distinct transport activity of tetraethylammonium from L-carnitine in rat renal brush-border membranes. AB - We investigated the contribution of the Na(+)/L-carnitine cotransporter in the transport of tetraethylammonium (TEA) by rat renal brush-border membrane vesicles. The transient uphill transport of L-carnitine was observed in the presence of a Na(+) gradient. The uptake of L-carnitine was of high affinity (K(m)=21 microM) and pH dependent. Various compounds such as TEA, cephaloridine, and p-chloromercuribenzene sulfonate (PCMBS) had potent inhibitory effects for L carnitine uptake. Therefore, we confirmed the Na(+)/L-carnitine cotransport activity in rat renal brush-border membranes. Levofloxacin and PCMBS showed different inhibitory effects for TEA and L-carnitine uptake. The presence of an outward H(+) gradient induced a marked stimulation of TEA uptake, whereas it induced no stimulation of L-carnitine uptake. Furthermore, unlabeled TEA preloaded in the vesicles markedly enhanced [14C]TEA uptake, but unlabeled L carnitine did not stimulate [14C]TEA uptake. These results suggest that transport of TEA across brush-border membranes is independent of the Na(+)/L-carnitine cotransport activity, and organic cation secretion across brush-border membranes is predominantly mediated by the H(+)/organic cation antiporter. PMID- 12543443 TI - Event-related potentials to Stroop and reverse Stroop stimuli. AB - In the Stroop task, the latency of response to a colour is either faster or slower in the presence of a congruent or incongruent colour-word (J. Exp. Psychol. 18 (1935) 643). Debate remains as to whether this effect occurs during early stimulus processing or late response competition. The present study examined the task using reaction time (RT) and event-related potentials to determine temporal differences in this processing. The 'reverse Stroop' effect (where colour interferes with processing of a colour-word) which is much less well established, was also examined. Standard Stroop interference was found as well as reverse Stroop interference. A late lateralised negativity at frontal sites was greater for Incongruent trials and also for the word-response (reverse Stroop) task, and was interpreted as semantic selection and word-rechecking effects. Late positive component latency effects generally mirrored the speed of processing of the different conditions found in RT data. Stroop effects were also found in early temporal N100 and parietal P100 components, which differentiated Congruent from Incongruent trials in the reverse Stroop but not the standard Stroop, and were interpreted as early perception of physical mismatch between the colour and word. It was concluded that Stroop stimuli are processed in parallel in a network of brain areas rather than a particular structure and that Stroop interference arises at the output stage. PMID- 12543444 TI - The regularities of the discrete nature of multi-variability of EEG spectral patterns. AB - The short-term structure of electroencephalogram (EEG) spectral transformations during different brain functional states (closed/opened eyes and memory task) was studied. It was shown that approximately 50% of spectral pattern (SP) types occur not more than 2-3 times per 149 analysis epochs in a 1-min EEG. The remaining 50% of SP types were the same for the different EEG channels, in all subjects and various brain functional states. Additionally, a high incidence of the neighboring SP types in strongly overlapping (by 80%) 2-s analysis epochs of the EEG was shown. The SP identified in a given epoch has only a limited predictive value on the SPs identified in the subsequent epochs. The incidence effect was restricted by the limited SP set and by a 50% reduction in the functionally active SPs, which resulted in a temporary stabilization of SPs in sequential combinations. The parameters of temporary stabilization of SPs were significantly different from 'random' EEG which provides evidence of the non-occasional character of stabilization of the main dynamic parameters of neuronal activity. Thus, the findings suggest that the multi-variability of neuronal nets is discrete in time, and limited by the dynamics of the short quasi-stable brain states. PMID- 12543445 TI - Predicting blood pressure reactivity and heart rate variability from mood state following coronary artery bypass surgery. AB - STUDY OBJECTIVES: Coronary Artery Bypass Graft (CABG) surgery is a common and successful procedure for revascularisation. However, the experience can induce emotional reactions prior to and following surgery. This study aimed to document changes in blood pressure (BP) reactivity and heart rate variability (HRV) following CABG surgery, and to determine the impact of mood state, particularly anxiety and depression upon cardiovascular functioning. METHOD: Twenty-two patients preparing to receive elective, first time CABG surgery were recruited from The Cardiothoracic Centre, Liverpool, UK and psychologically assessed using the Hospital Anxiety and Depression Scale (HAD), Global Mood Scale (GMS) and the Dispositional Resilience Scale (DRI). BP and heart rate responses were also measured during four conditions: baseline response; laboratory session; ambulatory monitoring; and self-initialised recordings during the ambulatory period. In addition, HRV was measured for 12 h in conjunction with the ambulatory monitoring period. All measures were assessed 1 week prior to surgery and 2 months following surgery. RESULTS: A significant decrease in negative mood and an increase in positive mood were reported following surgery. Forty percent of patients were clinically anxious and depressed prior to surgery although this was reduced to 27% after surgery. Depression was the strongest independent predictor of pre-operative BP and HRV whilst anxiety was most significantly related to follow-up BP reactivity. DBP was most strongly predicted by mood state. CONCLUSIONS: These results suggest that patients with higher levels of anxiety and depression are at risk of reduced HRV and increased BP reactivity in response to mental stressors. The study also strongly suggests that current patient services should be expanded to acknowledge the role of psychological factors within clinical prognosis after CABG surgery. PMID- 12543446 TI - An investigation into asymmetrical cortical regulation of salivary S-IgA in conscious man using transcranial magnetic stimulation. AB - Transcranial magnetic stimulation (TMS) can activate discrete areas of the cerebral cortex through the intact skull of healthy conscious volunteers. A magnetic coil generates a brief and focused magnetic field that penetrates the skull to activate the specific area of cerebral cortex beneath. This non-invasive procedure is painless, well tolerated by participants and now widely used to explore brain function. We used TMS to investigate asymmetrical cortical regulatory influences on one aspect of immune function: secretion of the antibody immunoglobulin A (S-IgA) into saliva. The right and left temporo-parietal occipital cortex of 16 healthy, conscious subjects was stimulated on two different occasions, at least 1 week apart. There was an immediate increase in S IgA concentration following both right and left stimulation. Saliva volume was reduced immediately post-right but not left stimulation. When secretion (microg/min) of S-IgA was calculated (controlling for changing saliva volume) an increase was apparent following left but not right hemisphere stimulation. Furthermore, a significant difference between the relationship between S-IgA concentration and volume of saliva post-left and right stimulation was observed. We conclude that TMS can be used as a tool to investigate cortical regulation of autonomic and immune function in healthy, conscious human subjects and that secretion of saliva and S-IgA is differentially affected by stimulation of the left and right cerebral cortex. PMID- 12543447 TI - Paradox lost? Exploring the role of alpha oscillations during externally vs. internally directed attention and the implications for idling and inhibition hypotheses. AB - Although slow waves of the electroencephalogram (EEG) have been associated with attentional processes, the functional significance of the alpha component in the EEG (8.1-12 Hz) remains uncertain. Conventionally, synchronisation in the alpha frequency range is taken to be a marker of cognitive inactivity, i.e. 'cortical idling'. However, it has been suggested that alpha may index the active inhibition of sensory information during internally directed attentional tasks such as mental imagery. More recently, this idea has been amended to encompass the notion of alpha synchronisation as a means of inhibition of non-task relevant cortical areas irrespective of the direction of attention. Here we test the adequacy of the one idling and two inhibition hypotheses about alpha. In two experiments we investigated the relation between alpha and internally vs. externally directed attention using mental imagery vs. sensory-intake paradigms. Results from both experiments showed a clear relationship between alpha and both attentional factors and increased task demands. At various scalp sites alpha amplitudes were greater during internally directed attention and during increased load, results incompatible with alpha reflecting cortical idling and more in keeping with suggestions of active inhibition necessary for internally driven mental operations. PMID- 12543448 TI - The effect of training distinct neurofeedback protocols on aspects of cognitive performance. AB - The use of neurofeedback as an operant conditioning paradigm has disclosed that participants are able to gain some control over particular aspects of their electroencephalogram (EEG). Based on the association between theta activity (4-7 Hz) and working memory performance, and sensorimotor rhythm (SMR) activity (12-15 Hz) and attentional processing, we investigated the possibility that training healthy individuals to enhance either of these frequencies would specifically influence a particular aspect of cognitive performance, relative to a non neurofeedback control-group. The results revealed that after eight sessions of neurofeedback the SMR-group were able to selectively enhance their SMR activity, as indexed by increased SMR/theta and SMR/beta ratios. In contrast, those trained to selectively enhance theta activity failed to exhibit any changes in their EEG. Furthermore, the SMR-group exhibited a significant and clear improvement in cued recall performance, using a semantic working memory task, and to a lesser extent showed improved accuracy of focused attentional processing using a 2-sequence continuous performance task. This suggests that normal healthy individuals can learn to increase a specific component of their EEG activity, and that such enhanced activity may facilitate semantic processing in a working memory task and to a lesser extent focused attention. We discuss possible mechanisms that could mediate such effects and indicate a number of directions for future research. PMID- 12543449 TI - Emotional sweating response in a patient with bilateral amygdala damage. AB - Sweat output on the palm or sole is distinct from thermoregulatory sweating and has been designated emotional sweating. The amygdala has been implicated in this phenomenon, but the role it plays remains unclear. We had the chance to evaluate emotional sweating in a 21-year-old female with bilateral restricted amygdala lesions caused by idiopathic subacute limbic encephalitis. At the peak of the illness, sweat responses in the palm were not evoked in this patient in conjunction with any sympathetic activation procedures, including deep inspiration, mental arithmetic, isotonic exercise, and tactile stimulation. After neurological improvement associated with diminution of amygdala lesions on the magnetic resonance imaging, normal sweat responses were incited. This indicates that the amygdala does play an important role in emotional sweating. PMID- 12543450 TI - Neotrofin increases heme oxygenase-1 selectively in neurons. AB - There is evidence that heme oxygenase plays a role in cellular defense against reactive oxygen species and thereby has neuroprotective effects. We examined the interaction of Neotrofin, a cognitive-enhancing and neuroprotective drug, with the heme oxygenase system. In adult rats, both a single administration or seven daily injections of Neotrofin at 10, 30 or 100 mg/kg intraperitoneally increased HO-1 immunoreactivity in neurons of the hippocampal formation and its connections including CA1-4, fornix, septal nuclei, hippocampal commissure, septohippocampal nucleus, fimbria, anteroventral thalamic nucleus, frontal and parietal cortex. Prominent HO-1 staining of neuronal cells in the proximity of blood vessels and circumventricular organs was also observed. Increasing doses of Neotrofin resulted in an increase in the number of neuronal populations expressing HO-1 with 100 mg/kg evoking a widespread neuronal cell response in brain. Quantification by ELISA confirmed that intraperitoneal administration of 100 mg/kg Neotrofin caused a significant increase in HO-1 protein expression in the hippocampus. The increase was evident by 6 h post-injection, peaked at 24 h with a 4-fold increase, and persisted for at least 48 h. Similarly, oral administration of 100 mg/kg Neotrofin produced a 5-fold increase in HO-1 protein 24 h post-administration. The effect of Neotrofin on HO-1 appears to be at the transcriptional level, as suggested by an increase in HO-1 mRNA levels. Neotrofin treatment was also associated with a significant increase in HO-2 mRNA levels in whole brain homogenate. These data may explain the neuroprotective effects of Neotrofin in models of excitotoxic neuronal injury. PMID- 12543451 TI - Heat acclimation prolongs the time to central nervous system oxygen toxicity in the rat. Possible involvement of HSP72. AB - Oxygen toxicity of the central nervous system (CNS-OT) can occur during diving with oxygen-enriched gas mixtures, or during hyperbaric medical treatment. CNS-OT is characterised by convulsions and sudden loss of consciousness, which may be fatal in diving. Heat acclimation is known to provide cross-tolerance to various forms of stress in different organs, including the brain. We hypothesised that heat acclimation may delay the onset of CNS-OT in the rat. Male Sprague-Dawley rats were acclimated to an ambient temperature of 32 degrees C for 4 weeks. Rats in the control group were kept at 24 degrees C. Both groups were exposed to oxygen at 608 kPa. EEG was recorded continuously until the appearance of the first electrical discharge preceding clinical convulsions. CO(2) production was measured simultaneously with the EEG. Latency to CNS-OT was measured and brain samples were taken for evaluation of heat shock protein 72 (HSP72) levels by Western blot analysis at the end of the acclimation period and during 4 weeks of deacclimation. Latency to CNS-OT was twice as long in the heat-acclimated rat, with insignificant changes in CO(2) production. This prolongation continued for 2 weeks during deacclimation. There was a significant increase in the level of HSP72 following heat acclimation, with a subsequent decrease during deacclimation. We conclude that heat acclimation prolongs latency to CNS-OT in a way that does not involve changes in metabolic rate. During deacclimation there was a linear relationship between latency to CNS oxygen toxicity and the level of HSP72. A possible beneficial effect of HSP72 is discussed. PMID- 12543452 TI - Cooperative expression of survival p-ERK and p-Akt signals in rat brain neurons after transient MCAO. AB - In order to determine possible coordinate expression of major survival signals, immunofluorescent analyses for phosphorylated ERK (p-ERK) and phosphorylated Akt (p-Akt) were carried out after 90 min of transient middle cerebral artery occlusion (MCAO) in rats. p-Akt single positive cells (E-/A+) were found in the sham control brains with weak signal intensity. The levels of both survival signals concurrently increased from 1 to 3 h after the reperfusion with the peak at 1 h, and the signals were much stronger in the ischemic penumbra (IP) than ischemic core (IC). The number of E-/A+ cells was larger in both the IC and IP than that of p-ERK single positive cells (E+/A-). The E+/A- cells were primarily expressed at 1 h in the IP. The number of p-ERK plus p-Akt double positive cells (E+/A+) peaked at 1 h, and the intensity was much stronger in the IP than IC. These findings suggest that p-ERK and p-Akt play independent roles, respectively as emergency or maintenance signal for survival at an early stage after reperfusion, and that both signals were cooperatively expressed especially in the IP. PMID- 12543453 TI - The efferent connections of the pupillary constriction area in the rat medial frontal cortex. AB - This study investigated, in the rat, the efferent projections of the pupillary constriction area, which is located within the medial frontal cortex. In order to identify the location of the pupillary constriction area, in preliminary experiments the medial frontal cortex was microstimulated. Intracortical microstimulation elicited pupillary constriction in a thin strip of cortex near the interhemispheric fissure and bordering the frontal eye field and vibrissae area of the somatomotor cortex. Seven animals received a single iontophoretic injection of Phaseolus vulgaris leucoagglutinin in the pupillary constriction area. In these cases, anterogradely labelled fibres and terminal-like elements were found in both hemispheres. The densest labeling was seen in several areas of the injected hemisphere, where labeled fibers prevailed in the secondary visual cortex. Dense labeled fibers were also found in the retrosplenial and cingulate cortex. In the thalamus, labeled fibers were seen in the intralaminar nuclei and posterior nuclear group. In the midbrain and pons, labeled fibers were located in the anterior pretectal area, superior colliculus and in the dorsolateral portion of the central gray. Contralaterally to the injection site, labeled fibers were distributed in the homotopic region. These findings led us to assume that, in the medial frontal cortex of the rat, besides controlling pupillary constriction, the pupillary constriction area may also be involved in controlling orientation and exploring behavior. PMID- 12543454 TI - Calmodulin and calmodulin-dependent kinase II mediate neuronal cell death induced by depolarization. AB - Depolarization has been known to play an important role in the neuronal damage that occurs following cerebral ischemia. In the present study, we investigated the roles of calmodulin (CaM) and CaM-dependent enzymes in depolarization-induced neuronal cell death. Treatment of primary cortical neurons with 10 microM veratridine, a voltage sensitive Na(+) channel activator, induced cell death as indicated by lactate dehydrogenase leakage from neurons. CaM antagonists (calmidazolium, trifluoperazine, W-7, and W-5) inhibited cell death induced by veratridine in a concentration-dependent manner. CaM kinase II (CaMKII) inhibitors (KN-62, KN-93, and myristoylated autocamtide-2 related inhibitory peptide), but not inhibitors of nitric oxide synthase or calcineurin, prevented veratridine-induced neuronal cell death. Veratridine rapidly activated CaMKII in neurons, and CaM antagonists and a CaMKII inhibitor suppressed the CaMKII activation. These results suggest that the CaM-CaMKII pathway contributes to depolarization-evoked cell death in neurons. PMID- 12543455 TI - D-(+)-glucose rescue against 1-methyl-4-phenylpyridinium toxicity through anaerobic glycolysis in neuroblastoma cells. AB - The active neurotoxin of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), 1 methyl-4-phenylpyridinium (MPP+), exerts its lethal effect by inhibiting Complex I of the electron transport chain (ETC). MPP+ shuts down aerobic oxidative phosphorylation and ETC-mediated ATP synthesis. The present investigation examines anaerobic survival during MPP+ toxicity in murine neuroblastoma cells Neuro 2-A (N2-A). MPP+ addition to the cells resulted in a reduction in cell viability, mitochondrial O(2) consumption (MOC) and ATP concentration in a dose dependent manner. However, the addition of 10 mM of D-(+)-glucose prevented MPP+ toxicity, attenuated the loss of ATP, but did not reverse the complete inhibition of MOC, indicating substrate level phosphorylation and explicit anaerobic survival. Glucose addition prevented MPP+-mediated drop in DeltaPsim, endoplasmic reticulum and intracellular organelle membrane potential tantamount to an increase of cell viability. Secondly, we examined the metabolic regulation of pyruvate dehydrogenase (PDH) and carnitine palmitoyl transferase (CPT) activities during glucose rescue. These enzymes exert control over acetyl CoA reservoirs in the mitochondria during aerobic metabolism. DL-6,8-Thioctic acid (PDH prosthetic group) and insulin slightly augmented metabolic rate, resulting in enhanced vulnerability to MPP+ in a glucose-limited environment. Additional glucose prevented these effects. Amiodarone (CPT inhibitor) and glucagon did not hamper or potentiate glucose rescue against MPP+. These data support strict anaerobic glucose utilization in the presence of toxic levels of MPP+. Moreover, the findings indicate that MPP+ exerts two distinct modes of toxicity (fast and slow death). With MPP+ (<1 mM), anaerobic glycolysis is operational, and toxicity is strictly dependent upon glucose depletion. MPP+ (1-10 mM) initiated acute metabolic collapse, with failure to sustain or switch to anaerobic glycolysis. In conclusion, overcoming energy failure against MPP+ may involve targeting rate limiting controls over anaerobic energy pathways. PMID- 12543457 TI - Rapid eye movement sleep induction by microinjection of the GABA-A antagonist bicuculline into the dorsal subcoeruleus area of the rat. AB - In cats, rapid eye movement sleep (REMS) can be induced rapidly and reliably by injections of the cholinergic agonist carbachol into the anterodorsal pontine tegmentum, also recognized as the perilocus coeruleus alpha, and designated the REMS Induction Zone (RIZ). In rats, the RIZ has been ascribed to a much larger and more ventral region within the entire oral pontine reticular formation. However, carbachol injections throughout this area produce only small, unreliable, and long latency REMS enhancements. The present study investigated whether REMS induction in the rat is possible by microinjection into the dorsal subcoeruleus nucleus (SubCD), a region with similarities to the cat RIZ. In freely moving unanaesthetized rats, microinjection of the GABA-A antagonist bicuculline significantly increased the amount and reduced the latency to REMS during a 2-h recording in the mid-light period. However, at effective doses, bicuculline usually also produced intermittent ipsiversive circling behavior that disrupted REMS maintenance. Attempts at eliminating this side-effect by: (i) coinjection of bicuculline with the NMDA antagonist, APV, (ii) lower bicuculline doses, or (iii) injection of the GABA-B antagonist, phaclofen, were unsuccessful. Other drugs injected into this area did not induce REMS; these included carbachol, the acetylcholinesterase inhibitor neostigmine, the glutamate agonist kainate, and vasopressin. In the rat, the SubCD is a highly sensitive region for both REMS induction and locomotor effects. PMID- 12543458 TI - A paired-pulse facilitation analysis of long-term synaptic depression at excitatory synapses in rat hippocampal CA1 and CA3 regions. AB - Paired-pulse facilitation (PPF) is a form of short-term, activity-dependent synaptic plasticity common to most chemically transmitting synapses, manifested as an enhancement in the amplitude of the second of two rapidly evoked excitatory postsynaptic potentials (EPSPs). The generally accepted explanation of PPF posits that residual intraterminal free [Ca(2+)] from the first action potential facilitates the probability of transmitter release evoked by the second stimulus. A common extension of this hypothesis postulates that any plastic change which alters the probability of transmitter release, should also alter the magnitude of PPF. In the present study, we examined the relationship between PPF and both stimulus- and chemically-evoked long-term depression of synaptic strength (LTD) at Schaffer collateral-CA1, commissural/associational-CA3 and mossy fiber-CA3 synapses in rat hippocampal slices. We observed no significant change in mean PPF associated with either electrically- or chemically-induced LTD at any of these synapses. However, a correlation analysis revealed a complex pattern of PPF changes with LTD, such that low initial PPF was correlated with increases in PPF, while high initial PPF was associated with decreases. Combined with previous findings supporting a presynaptic site for chemical and stimulus-evoked LTD, our current data suggests a complex set of neurosecretory modifications downstream of presynaptic Ca(2+) influx, may, at least in part, underlie the expression of LTD. PMID- 12543456 TI - Adenoviral GDNF gene transfer prevents motoneuron loss in the nucleus ambiguus. AB - We examined neuroprotective effects of an adenoviral vector encoding glial cell line-derived neurotrophic factor (AxCAhGDNF) on the lesioned adult rat motoneurons in the nucleus ambiguus. After vagal nerve avulsion, AxCAhGDNF, AxCALacZ (adenovirus encoding beta-galactosidase gene) or PBS was inoculated into the jugular foramen. Four days after the avulsion and treatment with AxCALacZ, the animals expressed beta-galactosidase activity in the lesioned motoneurons in the nucleus ambiguus. The animals avulsed and inoculated with AxCAhGDNF showed immunolabeling for GDNF in the nucleus ambiguus on the treated side and expression of virus-induced human GDNF mRNA transcripts in the brainstem tissue that contained the nucleus ambiguus of the treated side. The treatment with AxCAhGDNF after avulsion prevented the loss of lesioned motoneurons in the nucleus ambiguus, ameliorated the choline acetyltransferase immunoreactivity, and also suppressed the activity of nitric oxide synthase in these neurons. These results indicate that adenovirus-mediated GDNF gene transfer may prevent the degeneration of motoneurons in humans after either vagal nerve injury or recurrent laryngeal nerve injury. PMID- 12543459 TI - Subcellular identification of angiotensin I/II- and angiotensin II (AT1)-receptor immunoreactivity in the central nervous system of rats. AB - To gain insight into generating and transport mechanisms of angiotensin (ANG) in the brain the study was focused on the subcellular localization of ANG II and its AT(1)-receptors in the hypothalamus of rats. The present paper demonstrates ANG II- and AT(1)-receptor-immunolabelling at brain parenchyma vessels and at glial and neuronal structures in the perivascular region. Further, ANG II- and AT(1) receptor-immunoreactivity is shown at plasma membranes and intracellular structures in the ependyma of the third ventricle. Based upon a conventional horseradish peroxidase technique, combined with the classical substrate 3,3' diaminobenzidine, a procedure is introduced that will be useful with a variety of antibodies used on glutar- and paraformaldehyde-fixed brain tissue. This technique enables a fast correlation between light and electron microscopical results and might also provide an attractive alternative to colloidal gold labelling and silver-intensification techniques. PMID- 12543460 TI - Cytokines regulate c-Met expression in cultured astrocytes. AB - We investigated c-Met expression in cultured astrocytes and their regulation by cytokines. Immunocytochemistry revealed that c-Met was expressed in cultured astrocytes. Western blotting revealed that acidic and basic fibroblast growth factor (FGF) enhanced and hepatocyte growth factor (HGF) reduced c-Met expression. Reverse transcription-polymerase chain reaction revealed that FGFs and HGF enhanced c-met expression. These findings suggest that c-Met expressed in astrocytes may have important roles during the nervous system regeneration. PMID- 12543461 TI - Effects of acute hypotension on expression of cFos-like protein in the vestibular nuclei of rats. AB - The expression and regional distribution of cFos protein, which is an oncogene product and metabolic marker of neural excitation, were investigated in the vestibular nuclear complex following acute hypotension in adult Sprague-Dawley rats. Intravenous administration of nitroprusside elicited a 10-50% reduction in mean blood pressure for 10 min. Unilateral or bilateral chemical labyrinthectomies were performed 14 days before the start of the experiment to eliminate afferent signals from the peripheral vestibular receptors in the inner ear. All of the animals were sacrificed and the tissues were fixed 2 h after the onset of acute hypotension using the cardiac perfusion method for c-Fos immunohistochemical staining. The cFos-like immunoreactive (cFLI) neurons were expressed selectively in the central area of the medial vestibular nucleus following a 10% reduction in blood pressure. Once the blood pressure had fallen by 30%, bilateral expression of cFLI neurons was observed in the superior, medial, and spinal vestibular nuclei, but not in the lateral vestibular nucleus, of control rats with intact labyrinths. The expression of cFLI neurons increased proportionately with reductions in blood pressure. In unilaterally labyrinthectomized rats, acute hypotension induced the expression of cFLI neurons in vestibular nuclei contra lateral to the injured labyrinth, but not in the ipsilateral vestibular nuclei. However, cFLI neurons were not expressed in bilateral vestibular nuclei following acute hypotension in bilateral labyrinthectomized rats. These results suggest that afferent signals from the peripheral vestibular receptors are essential for cFos protein expression in the vestibular nuclei following acute hypotension. PMID- 12543462 TI - Involvement of protein kinase C in glutamate release from cultured microglia. AB - Glutamate release from microglial cells may cause neuronal damage. To elucidate the mechanism of glutamate release, we examined the possible regulation by nitric oxide and protein kinase C. Cultured microglia prepared from the whole brains of newborn rats released glutamate by the stimulation with lipopolysaccharide (LPS) dose dependently. The time course study revealed that glutamate release showed a long lag time about 6 h after LPS stimulation, whereas about 3 h lag time was observed in LPS-induced NO production. An inhibitor for NO synthase, N(G)-nitro-L arginine, could effectively inhibit the glutamate release. Glutamate release induced by LPS was enhanced by 1 nM phorbol myristate acetate (PMA). Furthermore, high concentrations of PMA (>10 nM) induced glutamate release even without LPS stimulation. Glutamate release stimulated either by 100 ng/ml LPS or 100 nM PMA was inhibited by staurosporine, and also by alpha-aminoadipate. These results provide insight into the pathways regulating microglial pathological activation by protein kinase C and may be a base for the protection against microglia-evoked neurotoxicity. PMID- 12543463 TI - Enhanced effects of central angiotensin II on cardiovascular and drinking responses in inbred polydipsic (STR/N) mice. AB - STR/N, an inbred strain of mice, is known to exhibit extreme polydipsia and polyuria. The objective of this study was to investigate the possible reasons for polydipsia. First, comparisons were made between STR/N mice and control mice from the Institute of Cancer Research (ICR) concerning daily drinking, urinary excretion, and basal cardiovascular function. Then, since angiotensin II (ANG II) is a potent stimulus for drinking behavior, we investigated the effects of intracerebroventricular (i.c.v.) administration of ANG II on cardiovascular and water intake responses. Daily water intake, food intake, urinary volume, and urinary electrolytes (Na and K) excretion were larger in STR/N mice than in ICR mice, and the basal blood pressure was significantly lower in STR/N mice than in ICR mice. The i.c.v. administration of ANG II (10 pmol/per mouse) resulted in increased mean arterial blood pressure (MAP) and water intake in both STR/N and ICR mice, but the changes in MAP were significantly larger in STR/N mice than in ICR mice. These results suggest that polydipsia in STR/N mice is at least partially attributable to high sensitivity of central ANG II receptors and low MAP. PMID- 12543464 TI - Capsaicin modulates K+ currents from dissociated rat taste receptor cells. AB - Chili pepper is one of most widely used spices. The main active component of chili pepper is the capsaicin. The effects of capsaicin on sensory nerve endings are well known; however, little is known regarding the direct effect of capsaicin on taste receptor cells (TRCs). In this study, patch clamp methods were used to study the effects of capsaicin on the K(+) currents in TRCs isolated from the rat circumvallate papilla. Fura-2 microspectrofluorimetry was also used to determine the effects of capsaicin on the intracellular Ca(2+) concentration ([Ca(2+)](i)). In the resting state, whole-cell experiments identified outward-rectifying K(+) currents, which were inhibited by 5 mM tetraethylammonium (TEA(+)) chloride. Voltage-dependent K(+) channels with a conductance of 55+/-4 pS (mean+/-S.E.M.; n=3), were observed in cell-attached patches. Capsaicin (500 nM) completely inhibited the outward-rectifying K(+) current in the whole-cell recordings. In cell-attached patches 500 nM capsaicin significantly reduced the open probability (P(o)) of the K(+) channels from 0.401+/-0.052 (n=3) in the resting state, to 0.018+/-0.002 (n=3, P<0.05 by unpaired t-test). In the fura-2-loaded TRCs, micromolar concentrations of capsaicin increased [Ca(2+)](i) in a dose-dependent manner, e.g., 100 microM capsaicin consistently increased the 340:380 fluorescence ratio from 1.04+/-0.05 in the resting state to 1.40+/-0.05 (n=28). These results suggest that capsaicin can enhance or modify the gustatory sensation by inhibiting the K(+) currents of the TRCs directly. PMID- 12543465 TI - Lipid rafts of purified mouse brain synaptosomes prepared with or without detergent reveal different lipid and protein domains. AB - Lipid rafts have been proposed to be important in a variety of functions including lipid transport, signal transduction and cell growth. There is increasing evidence that lipid rafts may play a role in cell functions in brain. Lipid rafts are typically isolated using a detergent such as Triton X-100. There has been, however, data from non-brain tissue indicating that preparation of lipid rafts using a detergent may represent different raft domains as compared with non-detergent preparation. The purpose of the present study was to compare protein and lipid markers of lipid rafts using a highly purified mouse synaptosomal fraction and non-detergent and detergent methods. The lipid raft marker proteins, alkaline phosphatase and flotillin, and the lipid markers, cholesterol and sphingomyelin, were highly enriched in lipid rafts prepared with detergent as compared with the non-detergent fraction. Enrichment of Na(+),K(+) ATPase was greater in the non-detergent lipid raft fraction as compared with lipid rafts prepared with detergent. Lipid rafts from the nerve terminal of neurons prepared with or without detergents may represent different membrane domains each with unique specialized functions. PMID- 12543466 TI - Transport of 5-formyltetrahydrofolate into primary cultured cerebellar granule cells. AB - Transport of 5-formyltetrahydrofolate (5-FTHF) into primary cultured cerebellar granule cells (CGC) was studied. Uptake of 5-FTHF into CGC was saturable with K(m)=2.86 microM and V(max)=40.8 pmol/mg protein/45 min in pH 7.4 medium. Uptake of 5-FTHF in the astrocytes has a similar style in the time curve. Uptake of 5 FTHF is characterized by countertransport because adding unlabeled 5-FTHF in the medium resulted in the efflux of labeled 5-FTHF. Uptake of 5-FTHF was inhibited by the structural analogs 5-methyltetrahydrofolate, methotrexate and folic acid (K(i)=6.64, 7.69, and 19.38 microM, respectively). Uptake was significantly decreased by high concentrations of sodium azide and sodium arsenate but not by sodium cyanide. Uptake was also inhibited by p-chloromercuriphenylsulfonate and by the anions probenecid and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid. Acute exposure of the cells to ethanol (100 mM) did not affect the uptake. It is concluded that CGC have a carrier-mediated system for the uptake of 5-FTHF and other folates. PMID- 12543467 TI - Effects of lidocaine reversible inactivation of the median raphe nucleus on long term potentiation and recurrent inhibition in the dentate gyrus of rat hippocampus. AB - Considering the fact that median raphe nucleus (MRN) constitutes one of the inputs of the hippocampus, the effects of reversible inactivation of MRN on long term potentiation (LTP) and recurrent inhibition in the dentate gyrus (DG) of rat hippocampus, in vivo, were examined. Rats were anesthetized with urethane (1.5 g/kg, i.p.). MRN was temporarily suppressed by intra-MRN injection of lidocaine (0.5 microl, 2%). For LTP induction, eight episodes of high frequency stimuli (100 Hz) were delivered to the perforant path (PP), each consisting of 10 stimuli at 100 Hz. Population spikes (PS) and population excitatory post synaptic potentials ((P)EPSP) in DG were recorded 10 min before, and 5, 10, 20, 40, 60 and 120 min after tetanization. MRN inactivation itself had no effect on the amplitude of baseline responses. The PS amplitude and (P)EPSP slope in rats, injected with intra-MRN lidocaine, 5 min before tetanization, were not different from the control group. However, at 120 min PS amplitude was significantly higher than control. Lidocaine injection 5 min after tetanic stimuli caused a significant decrease in PS amplitude (10, 20 and 60 min) and (P)EPSP slope (20 and 40 min) after tetanization. The data showed that inactivation of MRN has no effect on LTP induction in the DG of hippocampus but it does affect its maintenance, and this effect depends on the pre- or post-tetanic inactivation. In the last part of this study, in order to investigate the effect of MRN on the efficacy of recurrent inhibition in the perforant-dentate synapses, paired pulse was applied to the PP at 10 and 20 ms interpulse intervals. Inactivation of MRN increased the amount of recurrent inhibition in the DG with 20 ms interpulse interval. This observation indicates that MRN inhibits the recurrent inhibition mechanism, which is in accordance with the suggested role of MRN neurons on inhibition of hippocampal GABAergic interneurons. PMID- 12543468 TI - Functional consequences of the acute administration of the cannabinoid receptor antagonist, SR141716A, in cannabinoid-naive and -tolerant animals: a quantitative 2-[14C]deoxyglucose study. AB - Cannabinoid systems have been shown to be involved in the regulation of ingestive behaviors. Administration of the cannabinoid antagonist, SR141716A, markedly reduces intake of sucrose solutions, food pellets, and ethanol. The purpose of the present studies was to identify the neural substrates that mediate these actions in rats using the quantitative autoradiographic 2-[14C]deoxyglucose (2 DG) method. In the first study, rats were trained to lever press in daily 15-min sessions for food pellets under a fixed-ratio schedule of food presentation. On the day of the experiment, rats received SR141716A (0, 1 or 3 mg/kg, i.p.) 15 min prior to behavioral testing, and the 2-DG procedure was initiated immediately after the operant test session. The acute administration of SR141716A dose dependently decreased rates of responding and was accompanied by decreases in glucose utilization concentrated in the limbic system, particularly those areas mediating motivated behavior. Because the effects of SR141716A on behavior are intensified in animals tolerant to the effects of Delta(9)-THC, the purpose of the second study was to assess the effects of the SR141716A administration on food-maintained responding and rates of glucose utilization in tolerant animals. The suppression of responding was greater in tolerant than in drug-naive animals. Furthermore, decreases in cerebral metabolism were more intense and widespread. Although still concentrated in limbic regions, functional changes now included areas subserving the regulation of ingestive behavior including the hypothalamus. These data suggest that the effects of SR141716A administration shift in the tolerant animal and may involve different aspects of feeding behavior than in cannabinoid-naive animals. PMID- 12543469 TI - Lidocaine inactivation demonstrates a stronger role for central versus medial extended amygdala in medial forebrain bundle self-stimulation. AB - Given recent attention to the role of the extended amygdala (EA) in brain reward processes, this study examines the relative contributions of the medial versus central aspects of that forebrain macrostructure to the rewarding effects of medial forebrain bundle (MFB) stimulation. Thirty-one rats were self-stimulated at either the rostral or caudal MFB before and after lidocaine-induced inactivation of an EA target. Relative to non-injection baseline tests, the injection of 0.5 or 1.0 microl of 4% lidocaine into the central EA structures of the lateral bed nucleus of the stria terminalis, the central sublenticular EA, and the interstitial nucleus of the posterior limb of the anterior commissure frequently and substantially disrupted the rewarding effect of MFB stimulation, whereas comparable saline infusions did not. The effects were most pronounced when the central EA was inactivated either bilaterally or ipsilateral to the stimulation site. Contralateral inactivation was less effective but did impair the stimulation's reward effects in several cases. Inactivation of medial EA structures did not have as great or as consistent effects on stimulation reward value except when the lidocaine infusion encroached on the MFB itself. These results support prior demonstrations of the EA's role in brain reward and motivational processes and further show that the central rather than medial aspects of the EA are particularly relevant. The results are discussed in the context of possible anatomical substrates supporting MFB self-stimulation. PMID- 12543470 TI - Retinal lesions affect extracellular glutamate levels in sensory-deprived and remote non-deprived regions of cat area 17 as revealed by in vivo microdialysis. AB - This study aimed at gaining insight into the role of the excitatory neurotransmitter glutamate in topographic map reorganization in the sensory systems of adult mammals after restricted deafferentations. Hereto, in vivo microdialysis was used to sample extracellular glutamate from sensory-deprived and non-deprived visual cortex of adult awake cats 18 to 53 days after the induction of restricted binocular retinal lesions, and in topographically corresponding cortical regions of control animals. A microbore HPLC-ED method was applied for the analysis of the microdialysates. In normal subjects, the visual cortex subserving central and peripheral vision showed similar extracellular fluid glutamate concentrations. In contrast, in animals with homonymous central retinal lesions, the extracellular glutamate concentration was significantly lower in central, sensory-deprived cortex compared to peripheral, non-deprived cortex. Compared to control regions in normal subjects, glutamate decreased in the extracellular fluid of deprived cortex but increased significantly in remote non-deprived visual cortex. These results not only suggest an activity-dependent regulation of the glutamate levels in visual cortex but also imply a role for perilesional cortical regions in topographic map reorganization following sensory deafferentation. PMID- 12543471 TI - A spinal mechanism for the peripheral anti-inflammatory action of indomethacin. AB - In the present study, we evaluated the consequences of prostaglandin E(2) (PGE(2)) or indomethacin injection into the spinal cord, on a model of peripheral inflammatory edema. Male Wistar rats (200-250 g) received PGE(2) (10 and 100 ng), intrathecally, at 2, 15, 30, and 60 min before an intraplantar carrageenan (CG; 300 microg) injection into the right hindpaw. The developing edema was measured hourly after CG injection, and the groups injected with PGE(2) 30 and 60 min before CG, presented significant edema potentiation. On the other hand, indomethacin (0.3, 0.6, 1.2, 2.5, and 5.0 microg) given intrathecally 60 min before CG injection, inhibited edema formation dose-dependently. The indomethacin effect was not inhibited by aminoglutethimide, which suggests that it was independent of endogenous steroid production. In addition, intrathecally given PGE(2) (10 and 100 ng) dose-dependently reversed the anti-edematogenic effect of indomethacin given by the same route (2.5 microg, i.t.). This suggests that the anti-edematogenic effect produced by intrathecally given indomethacin is probably due to prostaglandin synthesis inhibition at the spinal cord. It is suggested here that during inflammation, prostaglandin may be released into the spinal cord potentiating dorsal root reflexes that contribute to the peripheral edema formation. The inhibition of this potentiation by indomethacin may be a mechanism embedded into the overall anti-inflammatory action of this drug. PMID- 12543472 TI - Effects of spinal administration of muscimol on C- and A-fibre evoked neuronal responses of spinal dorsal horn neurones in control and nerve injured rats. AB - Neuropathic pain is a common clinical problem with complex aetiology, mechanisms and symptoms. Alterations in spinal gamma-aminobutyric acid (GABA) receptors may contribute to persistent pain states. The aim of the present study is to investigate potential changes of spinal GABA(A)-receptor function following peripheral nerve injury. Effects of spinal administration of the GABA(A)-receptor agonist muscimol (0.1-30 microg/50 microl) on electrically-evoked responses of spinal neurones in control, spinal nerve ligated and sham operated halothane anaesthetised rats were studied. Spinal muscimol significantly (10 microg/50 microl) reduced evoked Abeta-, Adelta- and C-fibre responses of spinal neurones in control rats (58+/-22% of control, P<0.05; 3+/-2% of control, P<0.001; and 8+/ 7% of control, P<0.001; respectively). Muscimol produced significantly greater inhibition of Adelta- and C-fibre evoked neuronal responses compared to Abeta fibre evoked neuronal responses in control rats (P<0.001). C-fibre mediated post discharge responses and the non-potentiated C-fibre evoked responses were significantly inhibited by muscimol in control rats. Inhibitory effects of muscimol (10 microg/50 microl) were blocked by pre-application of spinal bicuculline (10 microg/50 microl). Following either sham surgery, or spinal nerve ligation, spinal muscimol inhibited Abeta-, Adelta- and C-fibre evoked responses of spinal neurones to a similar extent, however significant inhibitory effects on the post-discharge response were not observed in nerve injured rats. Our data demonstrate that GABA(A)-receptor control of Abeta- and Adelta-fibre evoked responses are not altered in nerve injured or sham operated rats, compared to control. However, following nerve injury we report a reduction in GABA(A) receptor control of C-fibre responses, in particular in relation to post discharge responses. PMID- 12543473 TI - Glial cells in the chicken optic tectum. AB - We mapped the distribution of the three neuroglial cells, oligodendrocytes, astrocytes and microglia, in the chicken optic tectum using their specific markers, transferrin binding protein (TfBP), glial fibrillary acidic protein (GFAP), and Ricinus communis agglutinin-1 (RCA-1), respectively. Neuroglial cells showed distinct distribution according to their cell types. While the astrocytes were mainly found in the stratum opticum (SO), stratum album centrale (SAC) and stratum fibrosum periventriculare (SFP), with their processes extending throughout the entire optic tectum region, the oligodendrocytes were mainly scattered in the SO, stratum griseum centrale (SGC) and SAC. In the case of the microglia, ramified cells were found in nearly all the layers, with the majority being present in the SAC. This is the first report demonstrating the distribution of glial cells in the chicken optic tectum, and these findings may present a basis for further study. PMID- 12543474 TI - Different effects of phencyclidine and methamphetamine on firing activity of medial prefrontal cortex neurons in freely moving rats. AB - The purpose of this study was to compare the effects of systemically administered MAP with those of phencyclidine (PCP), both of which induced comparable locomotor activity, on firing activity of medial prefrontal cortex (mPFC) neurons in freely moving rats. The results show that, unlike PCP, acutely administered MAP produced little changes in firing activity of mPFC neurons. PMID- 12543475 TI - Activation of accessory olfactory bulb neurons during copulatory behavior after deprivation of vomeronasal inputs in male rats. AB - We examined whether the vomeronasal organ (VNO) is the sole receptor for pheromonal cues for male sexual behavior. Males carrying surgical removal of the VNO (VNOx) mated with stimulus females as sham-operated males, with a comparable number of mounts but a prolonged latency for ejaculation. In sham-operated males, mating increased cFos immunoreactivity in the granule and mitral cell layers of the accessory olfactory bulb and in the medial amygdala. VNOx diminished baseline as well as mating-induced cFos in the granule cell layer and in the medial amygdala; VNOx had no effect on either basal or induced cFos immunoreactivity in the mitral cell layer. Thus, during mating encounter, cFos expression in the mitral cell layer did not depend on VNO inputs. The medial amygdala may be modulated by impulses other than of mitral cell origin. PMID- 12543476 TI - Dopamine and cyclic AMP-regulated phosphoprotein immunoreactive neurons are innervated by axon terminals immunopositive for vasoactive intestinal polypeptide in the bed nuclei of the stria terminalis and central nucleus of the amygdala. AB - The bed nuclei of the stria terminalis (BST) and the central nucleus of the amygdala (CeA) are highly heterogeneous structures, which play a central role in the modulation and/or regulation of stress responses. The oval nucleus of the anterior division of BST (BSTov) and the CeA exhibit several dopamine and cyclic AMP-regulated phosphoprotein (DARPP-32) immunoreactive (ir) neurons. It has been demonstrated that DARPP-32, if phosphorylated, can inhibit protein-phophatase-1, thereby controlling other neuropeptide/neurotransmitter actions. In addition, a dense network of vasoactive polypeptide (VIP) immunoreactive axon terminals was also observed here. VIP, via its receptors, increases intracellular cAMP levels, therefore it can play an important role in regulating the phosphorylation of DARPP-32. Since the localization of DARPP-32- and VIP-ir neuronal structures overlaps in the BSTov and CeA, the aim of this study was to investigate the possible synaptic innervation of DARPP-32-ir neurons by fiber terminals immunopositive for VIP, to provide anatomical evidence for the interaction between a neuropeptide and a phosphoprotein. In summary, this study for the first time demonstrated that VIP-ir axon terminals innervate DARPP-32 perikarya and dendrites in the BSTov and CeA, which play an important role in the central autonomic regulation of stress responses. In addition, morphological evidence for possible interaction between neuropeptides and phosphoproteins was also provided at the electron microscopic level. PMID- 12543477 TI - Estrogen enhances potassium-stimulated acetylcholine release in the rat hippocampus. AB - Short-term estrogen replacement has been shown to enhance basal forebrain cholinergic function. Whether or not long-term estrogen replacement can enhance basal forebrain cholinergic function has been questioned in light of recent studies showing that several cholinergic measures which are increased following short-term treatment are not increased following longer-term (>30 days) treatment. In the present study, in vivo microdialysis was used to assess the effects of continuous estradiol replacement on basal forebrain cholinergic function. Our data show that 6-7 weeks of continuous estradiol replacement significantly enhanced potassium-stimulated acetylcholine release in the hippocampus of ovariectomized rats, and that this effect was reversed following discontinuation of the estrogen treatment. These data are consistent with the hypothesis that estrogen-mediated effects on cholinergic neurons can contribute to the effects of long-term estrogen replacement on cognitive performance recently described. PMID- 12543478 TI - Trade-offs between social behaviour and foraging by sheep in heterogeneous pastures. AB - In heterogeneous pastures, groups of sheep may have to alter their social behaviour in order to graze patches of preferred vegetation. In this event, patch size, inter-patch distance and the contrast between patch and background vegetation are likely to affect behaviour. In this experiment, groups of five female Scottish Blackface sheep grazed for 2-h periods in 0.1ha grass plots containing seven 1.5mx1.5m patches of improved vegetation, with inter-patch distances of 1.5, 6 or 11.5m. Background vegetation was of either medium or poor quality. Control plots contained no patches. On average, sheep spent 44% of the time grazing patches, although patches comprised only 1.6% of the total plot area. Inter-patch distance did not affect accumulated time spent grazing patches during the first 30min, but patch residence time and the number of sheep on a patch increased with inter-patch distance. The distribution of nearest neighbour distances was altered when patches were 6 or 11.5m apart, compared to no patches. Accumulated time spent grazing patches and the number of sheep on a patch were greater with poor than medium backgrounds. Sheep visited patches frequently and for short periods and it is suggested that sheep often moved off patches as a result of competition. The results provide evidence that sheep make trade-offs between social and foraging behaviour and demonstrate the importance of interactions between social spacing and the size and spacing of vegetation patches, when sheep forage in heterogeneous pastures. PMID- 12543479 TI - Preexposure to the stimulus elements, but not training to detect them, retards human odour-taste learning. AB - Odours are judged to smell sweeter following simultaneous oral pairings with the tastant sucrose and sourer after parings with the tastant citric acid. This effect may result from human participants perceiving and encoding a unitary odour taste percept. This study examined two factors thought likely to disrupt such encoding; (a) preexposure to the mixture elements and (b) training to spot the elements of taste-odour mixtures. Half of the participants were trained to identify tastes and smells and half received no training. All participants were preexposed to two odours (A, B) and two tastes (X, Y), followed by pairings of these stimuli (AX, BY) and then by pairings between two non-preexposed odours and the same tastes (CX, DY). This process was then repeated on a second session. Odour-taste learning was retarded following preexposure, but was unaffected by training. These findings suggest; (1) that odour-taste mixtures may be cognitively impenetrable and (2) that preexposure leads to encoding of A and B, which are then resistant to interference when further pairings are presented (i.e. AX, BY). PMID- 12543480 TI - Lateralised behaviour in the domestic dog, Canis familiaris. AB - Paw use on three tasks in 53 domestic dogs (Canis familiaris) was studied to determine whether the animals exhibited a significant paw preference, and, if so, to explore the direction and strength of the dogs' preferred paw use, both within, and between, tasks. The influence of the dogs' sex on their paw preferences was also explored. The findings revealed that lateralised behaviour was strongly sex related. Male and female showed paw preferences at the level of the population but in opposite directions. Female dogs had a greater preference for using their right paw on all tasks, whilst males were more inclined to adopt their left paw. Analysis revealed significant positive relationships for two of a possible three intertask correlations. Overall, the findings suggest that there are two distinct populations of paw preference in the domestic dog that cluster around the animals' sex. It is recommended that further work be conducted to explore the influence of pre-training on dogs' paw preferences before generalisations regarding laterality in this species are drawn. PMID- 12543481 TI - The effect of non-contingent negative and positive reinforcement operations on the acquisition of superstitious behaviors. AB - The differential effects of non-contingent positive and negative reinforcement operations on the acquisition of superstitious behaviors and rules were investigated in two experiments. College students were instructed to try to produce and/or keep the word "GOOD" on a computer screen (positive reinforcement), or to try to prevent and/or remove the word "BAD" from the screen (negative reinforcement) using response keys. Data from both experiments indicated that participants exposed to lean schedules of negative reinforcers believed that they had greater control over non-contingent stimulus events than participants exposed to either rich or lean schedules of positive reinforcers. These findings and results from research investigating everyday superstitious activities suggest that, relative to positive reinforcement, negative reinforcement operations may provide a more fertile condition for the development and maintenance of superstitious behaviors. PMID- 12543482 TI - Manipulating decision processes in the human scalar timing system. AB - Two experiments attempted to manipulate the decision processes used in a temporal generalisation task with humans. In Experiment 1, payoffs (points awarded or deducted) were used to try to alter behaviour when the standard duration was 400ms, and the comparison durations ranged from 100 to 700ms in 100ms steps. Two conditions which either encouraged or discouraged the subject to identify a comparison duration as the standard were compared with a neutral condition. Encouraging identifications of the standard increased the proportion of identifications of the standard, whereas the discouraging manipulation had more ambiguous effects. Using the "modified Church and Gibbon" model, it was shown that the effect of the encourage manipulation was an increase in the response threshold, consistent with the information-processing version of scalar timing theory. A second experiment compared encourage and discourage manipulations with a more difficult discrimination (comparison durations spaced in 50ms steps around the 400ms standard), and with more distinct payoff differences for the different conditions. Behavioural effects were much more marked in Experiment 2, with the encourage condition producing more identifications of a comparison duration as the standard for all comparison durations except the shortest, compared with the discourage condition. Modelling showed that the main theoretical difference between the two conditions was in a change in the response threshold, in a manner consistent with the scalar timing model. PMID- 12543483 TI - Effects of noise on the performance of rats in an operant discrimination task. AB - Two experiments examined the effect of the presentation of an irregular, moderate intensity auditory stimulus ('noise') on the performance of rats in an operant discrimination task. In Experiment 1, rats first learned to press a lever in the presence of a visual stimulus but not in its absence. Discrimination performance was impaired during subsequent exposure to noise. In Experiment 2, different groups of rats learned the discrimination task under a noise or a no-noise condition. Thereafter, all rats were tested under each noise condition. Discrimination performance was best when the noise condition at test was identical to the noise condition at training. These results were discussed in the framework of arousal, distraction, generalization decrement, and contextual occasion setting. They point to the necessity of using a 2x2 factorial design in human and animal research on noise effects, with noise condition at training (noise present or absent) and noise condition at test (noise present or absent) as factors. PMID- 12543484 TI - Factorial correspondence analysis of fear-related behaviour traits in Japanese quail. AB - Factorial correspondence analysis was performed on 341 quails from a F2 cross between two lines divergently selected on the duration of tonic immobility over 29 generations. Several fear- or stress-related traits were recorded, i.e. tonic immobility duration, number of inductions needed to induce tonic immobility, open field behaviour (time spent walking, latency before first movement and number of defecations), asymmetry of tibia lengths and corticosterone concentration after restraint stress. Variables were categorised in classes and analysed by factorial correspondence analysis. The first axis was mostly described by open-field behaviour, and the second by tonic immobility traits (duration of tonic immobility and number of inductions), which showed that these behaviours were almost independent. No relationship was found between axes of the factorial correspondence analysis and corticosterone concentration or asymmetry of tibia lengths, showing that these variables reflected other characteristics of stress susceptibility than those described by tonic immobility and open-field behaviour. These results show that reaction to stress of quails is a multidimensional trait and cannot be summarised by one trait. PMID- 12543485 TI - How mothers find their pups in a colony of Antarctic fur seals. AB - In Antarctic fur seals, Arctocephalus gazella, mothers must identify their own young among hundreds or even thousands of pups, if they are to invest in their own offspring and avoid misdirecting their parental care. When returning to their breeding colony from a foraging trip of several days at sea, mothers have to find and identify their young before suckling can occur. There appears to be little confusion about which pup belongs to a mother, and adoption is absent or rare. Using behavioral observations, we investigated the means by which female Antarctic fur seals identified their pups in a breeding colony of about 750 mother-pup pairs on Kerguelen Island. We evaluated the importance of vision, scent communication, vocalizations, and rendezvous locations as possible explanations of how mothers find their pups. Every pup that a mother examined, whether her own or not, exchanged naso-nasal inspection with her, suggesting a strong role for olfactory communication in individual recognition. Both mothers and pups called to each other, and mothers that searched for pups over a longer period gave more calls and encountered more pups. Thus, vocalizations may have been used to attract pups that might be offspring. Nursing usually occurred in the same place from the end of one maternal visit to the colony and the arrival at the beginning of the next visit, suggesting that nursing locations may serve as a meeting place, or rendezvous, for mothers and pups. These results suggest that finding pups is a two-stage process for females, in which pups for sampling are attracted by calls or examined at the previous nursing location, and then individual identification is made by olfactory cues. PMID- 12543486 TI - The role of differential reinforcement in predator avoidance learning. AB - Little is known about how predator recognition develops under natural conditions. Predispositions to respond to some stimuli preferentially are likely to interact with the effects of experience. Convergent evidence from several studies suggests that predator-nai;ve tammar wallabies (Macropus eugenii) have some ability to respond to vertebrate predators differently from non-predators and that antipredator responses can be selectively enhanced by experience. Here, we examined the effects of differential reinforcement on responses to a model fox (Vulpes vulpes), cat (Felis catus) and conspecific wallaby. During training, tammars experienced paired presentations of a model fox and a simulated capture, as well as presentations of a wallaby and a cat alone. Training enhanced responses to the fox, relative to the conspecific wallaby, but acquired responses to the two predators did not differ, despite repeated, non-reinforced presentations of the cat. Results suggest that experience interacts with the wallabies' ability to perceive predators as a natural category. PMID- 12543487 TI - Homing in pelagic birds: a pilot experiment with white-chinned petrels released in the open sea. AB - During the breeding period white-chinned petrels (Procellaria aequinoctialis) repeatedly perform long foraging trips in the open ocean from their breeding island, and are able to home with an astonishing precision. The orientation mechanisms involved are not yet known. By analogy with those used by desert ants moving in a similarly "featureless" environment, one can hypothesise that petrels may home using path-integration. We displaced 11 white-chinned petrels 725-785km from their burrows to the open sea, preventing them from using visual and magnetic route-based information. Three birds carried satellite transmitters. Our results showed that they can home rather efficiently in such conditions. PMID- 12543488 TI - The importance of stereoselective determination of drugs in the clinical laboratory. AB - About 56% of the drugs currently in use are chiral compounds, and 88% of these chiral synthetic drugs are used therapeutically as racemates. Only a few of these drugs qualify for a stereospecific determination in a clinical laboratory for therapeutic drug monitoring of patients. If the qualitative and quantitative pharmacokinetic and pharmacodynamic effects are similar, the enantiomers do not need to be separated. However, if the metabolism of the different stereoisomers is handled by different enzymes which are either polymorphic or can be induced or inhibited, and if their pharmacodynamic effects have differences either in strength or in quality, enantiospecific analysis is urgently needed. Unfortunately, there are many racemic drugs where the stereospecificity of the metabolism and/or the pharmacodynamic effects of the enantiomers is not known today. For these drugs, there is a great need for studies concentrating on these differences to improve treatment of the patients. PMID- 12543489 TI - Preparative chiral chromatographic resolution of enantiomers in drug discovery. AB - A brief account is given of the role of preparative chromatography for the direct separation of enantiomers in the drug discovery process. Although it is not yet possible to predict the outcome of a chromatographic resolution attempt, and the optimisation procedure sometimes might be time-consuming, the technique is still indispensable as a means to obtain both enantiomers in pure form from a drug racemate for biological testing. The most suitable types of chiral stationary phases (CSPs) available for this purpose are discussed with special reference to loadability and compatibility with different mobile phase systems. PMID- 12543490 TI - Resolution of chiral drugs by liquid chromatography based upon diastereomer formation with chiral derivatization reagents. AB - Chiral derivatization reagents for resolution of biologically important compounds, such as chiral drugs by high-performance liquid chromatography (HPLC), based upon pre-column derivatization and diastereomer formation, are reviewed. The derivatization reagents for various functional groups, i.e., amine, carboxyl, carbonyl, hydroxyl and thiol, are evaluated in terms of reactivity, stability, wavelength, handling, versatility, sensitivity, and selectivity. The applicability of the reagents to the analyses of drugs and bioactive compounds are included in the text. PMID- 12543491 TI - Enantioselective analytical methods in pharmacokinetics with specific reference to genetic polymorphic metabolism. AB - The new trend towards developing enantiospecific drugs has increased the interest in enantiospecific pharmacokinetics of chiral drugs, mainly in the case where only one of the two enantiomers is responsible for the pharmacological activity. Enantiospecific bioassays are also useful in investigating the pharmacokinetic behaviour of the two enantiomers when a given drug is marketed as racemate. The stability of the stereogenic centre in vitro and in vivo, as far as unidirectional and bidirectional inversions are concerned, is another reason for requiring stereospecific assay and bioassay. These assays are often complicated in order to achieve quantification, mainly for in vivo measurements, which are often in the low pg/ml range. This paper considers the enantiospecific bioassays, the methods and approaches used, the need for chemical derivatization, and the difficulties involved. It includes a specific discussion for the genetic polymorphic metabolism involving stereogenic centres. PMID- 12543492 TI - Species-dependency in chiral-drug recognition of serum albumin studied by chromatographic methods. AB - Stereoselective binding of benzodiazepine and coumarin drugs to serum albumin from human and six mammalian species were studied by chiral chromatographic techniques. The applied methods were affinity chromatography on the albumins immobilized on Sepharose 4B, high-performance liquid chromatography (HPLC) separation on columns based on human serum albumin (HSA) and bovine serum albumin (BSA), and chiral HPLC analysis of ultrafiltrates of solutions containing the racemic drug and the native protein. Substantial differences in preferred configurations and conformations were detected among the species. The binding stereoselectivity of the 2,3-benzodiazepine drug, tofisopam, in human, is opposite to that in all other species. In the binding of 1,4-benzodiazepines, dog albumin is very similar to HSA. Highly preferred binding of (S)-phenprocoumon was found with dog albumin. PMID- 12543493 TI - Simulated moving bed chromatography with supercritical fluids for the resolution of bi-naphthol enantiomers and phytol isomers. AB - The combination of the simulated moving bed (SMB) technique with supercritical fluid chromatography (SFC) leads to a process with unique features. Besides the known advantages of the SMB process, the use of supercritical carbon dioxide as the mobile phase offers the advantages of reduction in organic solvents and an easy eluent/solute separation. Because of the low viscosity and high diffusion coefficients of supercritical fluids, a high efficiency is possible. The steps of process development for SMB SFC are presented using the separations of the bi naphthol enantiomers and phytol isomers as examples. The development of a packed column SFC method at an analytical scale is shown for the separation of the bi naphthol enantiomers on a chiral stationary phase and CO(2) with a modifier as the mobile phase. The influence of the modifier, modifier content, and column configuration on productivity of the SMB SFC process was investigated by simulation. The first set of experiments was performed in the SMB separation of phytol isomers at low concentration to test the feasibility of the SMB SFC high purity separation of the binary mixtures. In the second set of experiments, the productivity of the process was increased by increasing the feed concentration up to 54 grams feed per liter stationary phase (SP) and hour (g(feed)/l(SP) h). PMID- 12543494 TI - Analysis of enantiomers of chiral phenethylamine drugs by capillary gas chromatography/mass spectrometry/flame-ionization detection and pre-column chiral derivatization. AB - Several important chiral phenethylamine agents such as mexiletine, fenfluramine, amphetamine, methamphetamine and N-n-propylamphetamine show stereoselective disposition in humans and large differences in therapeutic relevance and toxicity. To analyze the enantiomers of chiral amine drugs, stereoselective methods were developed to separate those enantiomers on an achiral capillary gas chromatography by pre-column chiral derivatization with S-(-)-N-(fluoroacyl) prolyl chloride. The stereoselectivity and sensitivity can be improved by chiral derivatization. The methods established offer enantioselective, simple, flexible and economic approaches for the analysis of chiral amine drug enantiomers in biological fluids. The methods have been used to determine S-(+)-methamphetamine in human forensic samples and to analyze enantiomers of amphetamine and fenfluramine in rat liver microsomes. PMID- 12543495 TI - Enantiomeric composition of abused amine drugs: chromatographic methods of analysis and data interpretation. PMID- 12543496 TI - Strategies for the enantiomeric determination of amphetamine and related compounds by liquid chromatography. AB - This paper summarizes recent research on the stereospecific analysis of amphetamine, its analogs and metabolites, by liquid chromatography. The different methods proposed have been evaluated and compared in terms of resolution power, time of analysis, sensitivity, or potential for automation. Chiral derivatization, followed by separation of the diastereomers formed in achiral chromatographic systems, is still the method preferred for the analysis of amphetamines at trace levels, as derivatization also improves analyte detectability. This is the method of choice for the enantiomeric analysis of amphetamines at the low concentrations typically encountered in biological samples. In recent years, special attention has been devoted to the development of alternatives for the automation of the analytical process by integrating the derivatization step into the chromatographic scheme. A promising alternative is the employment of beta-cyclodextrins as chiral selectors, both immobilized on the stationary phase and added to the mobile phase. However, with a few exceptions, beta-cyclodextrins perform better for non-derivatized amphetamines. Therefore, the utility of these selectors in the analysis of biological samples is limited. The reliability of less-used chiral stationary phases (Pirkle type, cellulose based or protein based), as well as methods based on the mathematical treatment of the chromatographic signal, are also discussed. PMID- 12543497 TI - Enantiomeric separations of amino alcohols by packed-column SFC on Hypercarb with L-(+)-tartaric acid as chiral selector. AB - The use of L-(+)-tartaric acid as a chiral mobile phase additive (CMPA) has been investigated in a packed-column SFC system. The CMPA, carbon dioxide, and methanol, containing a high concentration of aliphatic amine additive, were used as the mobile phase and Hypercarb as support [Gyllenhaal O., Karlsson A., SFC of metoprolol and other amino alcohols on Hypercarb (in preparation)]. Good enantioselectivities were obtained for tertiary amine homologues of 2-amino alcohols, used as beta-adrenoreceptor-blocking drugs. Moderate selectivities were observed for aromatic compounds having a second substituent in the ortho position. The overall retention was influenced by the aromaticity of the analytes as well as the presence of free electron pairs in the molecule. Increased concentrations of CMPA gave higher retention and also increased the enantioselectivity. The practical utility of this present enantioselective system was demonstrated on one batch of (S)-metoprolol that was N-methylated with methyl iodide. The enantiomeric separation was accomplished within 10 min. PMID- 12543498 TI - Chiral recognition of terpenoids in some pharmaceuticals derived from natural sources. AB - Capillary gas chromatography was applied to explore the enantiomeric composition of some terpenoids in pharmaceuticals of natural origin. The drugs under investigation were produced in Germany (Rowachol, Rowatinex), Poland (Terpichol, Terpinex, Rub arom, Herbolen and Oleum Camphoratum), Slovenia (Uroterp, Mentoklar) and United Kingdom (Olbas oil, Vicks Vapo Rub). The model compounds tested were: menthone, isomenthone, menthol, fenchone, borneol and camphor. It has been found that depending on the manufacturer, pharmaceuticals possessing similar chemical composition may differ considerably in enantiomer composition. Exceptionally large discrepancies have been found for the content of borneol and fenchone enantiomers in pharmaceuticals applied in liver and kidney diseases. It seems that the changes in enantiomeric composition are the main reason of the lack of general acceptance of natural medicines by clinicians. The study of enantiomeric composition may sometimes lead to information concerning the origin of preparation, i.e. natural or synthetic. PMID- 12543499 TI - Stereoselective chromatography of cardiovascular drugs: an update. AB - This review reports the latest achievements in chromatographic enantioseparations of various classes of cardiovascular drugs and selected applications of these methods in pharmaceutical and clinical analysis. The use of these drugs as test compounds for new chiral stationary phases and different parameters of chromatographic processes is also presented. PMID- 12543500 TI - High-performance liquid chromatographic separation of fluoroquinolone enantiomers: a review. AB - Fluoroquinolones are antibacterial agents widely used clinically. In recent years, there has been an important development of new derivatives, and more than 7000 analogues have been described today. Different fluoroquinolones (FQ) have one or two chiral centers in their chemical structure and are available as racemates, diastereoisomers, or pure enantiomers. The clinical and pharmaceutical uses of these compounds need effective analytical procedures for quality control and pharmacodynamic and pharmacokinetic studies. This review article focuses on the high-performance liquid chromatographic separation of fluoroquinolone stereoisomers by the use of derivatization methods and ligand exchange (LE) or chiral liquid chromatography. PMID- 12543501 TI - Liquid chromatographic separation of the enantiomers of fluoroquinolone antibacterials on a chiral stationary phase based on a chiral crown ether. AB - A chiral stationary phase (CSP) recently developed by bonding (diphenyl substituted 1,1'-binaphthyl) crown ether to silica gel for the liquid chromatographic separation of enantiomers was applied to the resolution of investigational fluoroquinolone antibacterial agents including gemifloxacin (formerly LB20304a). All fluoroquinolone compounds used in this study were resolved quite well on the CSP. Especially, the resolution of gemifloxacin and its analogs on the CSP was excellent and even greater than that on the commercial Crownpak CR(+). The resolution was found to be dependent on the type and the content of organic, acidic, and inorganic modifiers added to the mobile phase and on the column temperature. PMID- 12543502 TI - Separation of antifungal chiral drugs by SFC and HPLC: a comparative study. AB - The enantiomeric separation of several compounds, including an antifungal drug and several of its precursors, using HPLC and SFC is described in this work. The columns employed were based on polysaccharide derivatives and the results show that most of the separations obtained by SFC are better, in terms of high resolution and short analysis time, than those obtained by HPLC. Only one compound could not be resolved using SFC but, in this case, HPLC provided baseline resolution. PMID- 12543503 TI - Stereoselective determination and pharmacokinetics of dihydropyridines: an updated review. AB - All dihydropyridines, except nifedipine, have at least one chiral center, and their pharmacokinetics and clinical effects differ from one enantiomer to another. Chiral separation methods for dihydropyridines using chromatographic techniques are discussed. The stereoselective pharmacokinetics of dihydropyridine calcium antagonists were reviewed in detail in 1995. The present review article updates the methods for the stereoselective determination of dihydropyridines using chromatographic techniques and summarizes the pharmacokinetics of the dihydropyridines, including the newest drugs under development. PMID- 12543504 TI - Enantiomeric separation of substituted 2-arylindoles on derivatised polysaccharide chiral stationary phases. AB - The enantiomeric separation of a series of 2-arylindoles, developed as 5HT(2A) receptor antagonists for the treatment of schizophrenia, was investigated. Evaluation of a number of chiral stationary phases (CSPs) suggested that Chiralcel OD-H and Chiralpak AD were the most versatile for these compounds, and were employed for more detailed studies. A degree of complementarity between the CSPs was observed, such that Chiralcel OD-H was more effective for piperidine containing molecules and Chiralpak AD for piperazine- and morpholine-containing molecules. The presence of a basic secondary amine was detrimental chromatographically, but resolution was improved substantially by employing diethylamine (DEA) in the mobile phase. All separations were either enthalpy controlled or showed no temperature dependence. Differential temperature effects between series highlighted the possibility of multiple binding modes on these CSPs. Based on this study, it is possible to make a more rational selection of chromatographic conditions for future novel analogues. PMID- 12543505 TI - Separation of oxazepam, lorazepam, and temazepam enantiomers by HPLC on a derivatized cyclodextrin-bonded phase: application to the determination of oxazepam in plasma. AB - The enantioselective high-performance liquid chromatography (HPLC) of three racemic 3-hydroxybenzodiazepines, oxazepam (Oxa), lorazepam (Lor), and temazepam (Tem), is a difficult operation because of the spontaneous chiral inversion in polar solvent. To solve this problem, we have developed an HPLC method based on a chiral Cyclobond I-2000 RSP column, maintained at 12 degrees C, and a reversed mobile phase (acetonitrile in 1% triethylamine acetate buffer, TEAA) at a flow rate of 0.4 ml/min. Peaks were detected by a photodiode-array detector at 230 nm for quantification and by an optical rotation detector for identification of (+) and (-) enantiomers. The results showed that peak resolutions of Oxa, Lor, and Tem enantiomers, analyzed under the same conditions, were 3.2, 2.0, and 1.8, respectively. For the determination of Oxa enantiomers in plasma of rabbits, extraction with diethyl ether at pH 1.5, a polar organic mobile phase, and a Cyclobond I-2000 SP column were used. Other analytical conditions were the same as previously described. Blood samples were immediately cooled at 4 degrees C and centrifuged at 0 degrees C for the collection of plasma. The results showed a difference in plasma S(+)- and R(-)-oxazepam concentrations in rabbits. No racemization of S(+)- or R(-)-Oxa enantiomers, added alone to blank plasma, was observed after extraction and enantioselective HPLC analysis. PMID- 12543506 TI - Direct calculation and computer simulation of the enantiomerization barrier of oxazepam in dynamic HPLC experiments--a comparative study. AB - Dynamic chromatographic methods constitute a versatile approach to the rapid and precise determination of enantiomerization barriers of stereolabile drugs. In the present study enantioselective dynamic high-performance liquid chromatography (DHPLC) was employed to determine the enantiomerization barrier of oxazepam. Dynamic elution profiles, exhibiting plateau formation and/or peak broadening between 20 and 60 degrees C at pH 2.6 and pH 8 were obtained in the presence of the chiral stationary phase (CSP) Nucleodex-beta-PM (permethylated beta cyclodextrin chemically bonded to silica) using a 6:4 mixture of phosphate buffer and methanol as mobile phase. Evaluation of the experimental chromatograms was performed by the novel approximation function (AF) (without computer simulation), and by the stochastic model implemented in the ChromWin simulation software (with computer simulation) furnishing the respective apparent forward rate constants, k(1)(app)(T). From the rate constants, k(1)(app)(T), measured at variable temperatures, the kinetic Eyring activation parameters, deltaG(T)(#), deltaH(#) and deltaS(#), of the enantiomerization of oxazepam were obtained. By variation of the flow rate of the mobile phase, the expected independence of the enantiomerization barrier from the chromatographic time scale was demonstrated for the first time. PMID- 12543507 TI - High-performance liquid-chromatographic determination of warfarin enantiomers in plasma with automated on-line sample enrichment. AB - A sensitive and selective chiral high performance liquid chromatographic method was developed for the direct determination of R- and S-warfarin enantiomers in human plasma. The method involved direct injection of human plasma onto a semipermeable surface (SPS) guard column, washing the proteins from the column with aqueous acetonitrile and back flushing the analytes onto a reversed phase ovomucoid silica HPLC column using switching valves. After separation, the analytes were simultaneously detected and quantitated with a fluorometer. The recoveries of R-warfarin from human plasma at 25 and 2500 ng/ml were 98.9% and 88.1%, respectively. The recoveries of S-warfarin at 25 and 2500 ng/ml were 105.4% and 93.9%, respectively. Using 100 microl of human plasma, the lower limit of quantification for both R- and S-warfarins was 25 ng/ml. Linear responses in analyte/internal standard peak height ratios were observed for analyte concentrations ranging from 25 to 2500 ng/ml for both enantiomers. Fluorescence chromatograms of drug-free human plasma showed no interfering peaks with retention times similar to those for R- and S-warfarins and the internal standard. Results from a 3-day validation study for both enantiomers demonstrated excellent precision (1.7-9.0%) and accuracy (97-109%) across the calibration range. PMID- 12543508 TI - Insight into the chiral recognition of warfarin enantiomers by epichlorhydrin/beta-cyclodextrin polymer-based supports: determination of stoichiometry and stability of warfarin/beta-cyclodextrin polymer complexes. AB - The complexation of warfarin (W) enantiomers by a hydrosoluble high-molecular weight beta-cyclodextrin/epichlohydrin polymer (EP/beta-CD polymer) was studied using HPLC with a mobile phase of methanol/0.1 M Na acetate/acetic acid (pH 4) at 22 degrees C. It was found that the complexes (W/beta-CD unit) have a 1:1 stoichiometry. The stability constants of the complex involving each enantiomer and the polymer beta-CD units were determined in the mobile phase, and the highest stability of the complex (S-warfarin/beta-CD unit) was observed. From the chromatographic separations of warfarin enantiomers on different beta-CD or its derivative supports, we have deduced the role of the simultaneous presence of several glyceryl (-O-CH(2)-CHOH-CH(2)-O-) and dihydroxypropyl (-O-CH(2)-CHOH CH(2)OH) groups on one beta-CD ring in promoting the chiral recognition of warfarin enantiomers. PMID- 12543509 TI - Separation of albendazole sulfoxide enantiomers by chiral supercritical-fluid chromatography. AB - The enantioseparation of albendazole sulfoxide (ABZSO) by chiral supercritical fluid chromatography (SFC) on two columns, based on the polysaccharide derivatives Chiralpak AD and Chiralcel OD, was studied. The effect of different modifiers, methanol, ethanol, 2-propanol, and acetonitrile, was examined. The results showed that ABZSO can be separated on both columns, using an alcohol-type modifier. Using the Chiralpak AD column, the best results were obtained with 2 propanol and, in the case of the Chiralcel OD, with methanol. PMID- 12543510 TI - Effect of temperature on the reversal in the retention order of the enantiomers of mosapride on Chiral-AGP. AB - The retention order of the enantiomers of mosapride could be controlled by column temperature and mobile phase pH. In the presented paper, temperature studies have been used to study the thermodynamics of the reversal in retention order. A linear relationship was obtained plotting the logarithm of the capacity factor versus the inverted column temperature. However, at higher mobile phase pHs, the logarithm of the separation factor versus the inverted column temperature showed a non-linear behaviour and at the highest mobile phase pH used (pH=7.4), an optimum in the separation factor was observed. The plots showed that the thermodynamics for the two enantiomers of mosapride differ in the studied mobile phase pH interval. Thermodynamic values, enthalpy and entropy were calculated and showed that at a low mobile phase pH, the enantiomeric resolution was caused by differences in enthalpy between the two enantiomers. However, at a higher mobile phase pH, the chiral discrimination was a result of entropy effects. High correlation was obtained between experimental and predicted separation factors at different mobile phase pHs. PMID- 12543511 TI - Enantiomeric determination of amlodipine in human plasma by liquid chromatography coupled to tandem mass spectrometry. AB - A sensitive method for the separation and determination of amlodipine enantiomers in plasma has been developed based on solid-phase extraction (SPE) with disposable extraction cartridges (DECs) in combination with chiral liquid chromatography (LC). The SPE technique is used to isolate the drug from the biological matrix and to prepare a cleaner sample before injection and analysis by HPLC coupled to mass spectrometry. The DEC is filled with ethyl silica (50 mg) and is first conditioned with a 2.5% ammonia in methanol solution and then with ammonium acetate buffer. A 1.0-ml volume of plasma is then applied on the DEC. The washing step is first performed with ammonium acetate buffer and secondly with a mixture of water and methanol (65:35, v/v), while the final elution step is obtained by dispensing methanol containing 2.5% of ammonia. The eluate is then collected and evaporated to dryness before being dissolved in the LC mobile phase and injected into the LC system. The stereoselective analysis of amlodipine is achieved on a Chiral AGP column containing alpha(1)-acid glycoprotein as chiral selector by using a mobile phase consisting of a 10-mM acetate buffer (pH 4.5) and 1-propanol (99:1, v/v). The LC system is coupled to tandem mass spectrometry with an APCI interface in the positive-ion mode. The chromatographed analytes are detected in the selected reaction monitoring mode (SRM). The MS/MS ion transitions monitored are 409 to 238 for amlodipine, and 260 to 116 for S-(-) propranolol used as internal standard (IS). The method was validated considering different parameters, such as linearity, precision and accuracy. The limit of quantitation was found to be 0.1 ng/ml for each amlodipine enantiomer. PMID- 12543512 TI - Chiral reversed phase high-performance liquid chromatography for determining propranolol enantiomers in transgenic Chinese hamster CHL cell lines expressing human cytochrome P450. AB - An enantioselective assay for S-(-)- and R-(+)-propranolol in transgenic Chinese hamster CHL cell lines, expressing human cytochrome P450 (CYP), was developed. The method involves extraction of propranolol from the S(9) incubates, using S (+)-propafenone as internal standard, chiral derivatization with 2,3,4,6-tetra-O beta-D-glucopranosyl isothiocyanate and quantitation by reversed phase high performance liquid chromatography system with UV detection (lambda=220 nm). A baseline separation of propranolol enantiomers was achieved on a 5-microm reverse phase ODS column, with a mixture of methanol/water/glacial acetic acid (67:33:0.05, v/v) as mobile phase. The assay is linear from 5 to 500 microM for each enantiomer. The analytical method affords average recoveries of 99.2% and 98.8% for S-(-)- and R-(+)-propranolol, respectively. The limit of quantitation for the method is 5 microM for both S-(-)- and R-(+)-propranolol. The reproducibility of the assay is satisfactory (RSD < 10%). The method allowed study of the depletion of S-(-)- and R-(+)-propranolol in transgenic Chinese hamster CHL cell lines expressing CYP3A4, CYP2C18 and CYP2C9. PMID- 12543513 TI - Central composite design as a powerful optimisation technique for enantioresolution of the rac-11-dihydrooracin--the principal metabolite of the potential cytostatic drug oracin. AB - Three types of chiral stationary phase were used to achieve chromatographic resolution of enantiomers of rac-11-dihydrooracin (DHO), the principal metabolite of a potential cytostatic drug oracin. Chiralcel OD-R as a chiral stationary phase with mobile phase comprising acetonitrile (modifier) and sodium perchlorate (buffering component) proved to be the most suitable system. Chemometric optimisation based on the Box-Wilson central composite design was employed to find the optimum resolution. The optimum factor space was defined by three parameters: temperature, modifier concentration and buffer concentration. Newly designed chromatographic response functions based on a combination of resolution R(S) and retention time of the last component eluted t(RL) were employed to evaluate the resolution with regard to quality and quantity. Optimum values predicted from those models of response surfaces were in excellent agreement with the experimental results. The chromatographic resolution of DHO enantiomers is suitable for xenobiochemical studies on stereoselectivity and stereospecificity of biotransformation enzymes. PMID- 12543514 TI - Reversed-phase liquid chromatography of lisinopril conformers. AB - The effect of flow rate, temperature, pH, organic solvent and counter ion on peak shape and separation of the cis and trans conformers of lisinopril are investigated by HPLC. It was demonstrated that complete separation of the two isomers can be achieved at low temperature at either neutral or low pH together with appropriate type and concentration of organic solvent, whereas the elution of lisinopril as a single peak is favored by a decrease of flow rate, elevated temperature, choice of organic solvent (type and amount) and the use of an appropriate counter ion concentration. PMID- 12543515 TI - Effect of acidity on the enantiomeric resolution of thyroxine and tocainide by HPLC on a (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid column. AB - Enantiomeric resolution of thyroxine and tocainide was achieved on a (+)-(18 crown-6)-2,3,11,12-tetracarboxylic acid column. The mobile phases were methanol/water (4:1, v/v) and methanol/water containing 5 mM sulfuric acid (4:1, v/v) for tocainide and thyroxine respectively. The flow rate was 0.5 ml/min. The effect of the acidity on the chiral resolution of these drugs was studied. Detection was at 220 nm for both drugs. The values of alpha and Rs were 2.08-3.11 and 1.00-2.60, respectively, for thyroxine while the values of alpha and Rs were 1.13-1.26 and 0.10-1.30, respectively, for tocainide. PMID- 12543516 TI - Enantiomeric resolution of a novel chiral cannabinoid receptor ligand. AB - The enantiomeric resolution of a racemic novel cannabinoid receptor ligand conformationally restricted at the southern aliphatic chain was accomplished using a ChiralPak AD column. Both enantiomers were tested for their competitive binding to the rat brain CB1, mouse spleen CB2 and human CB2 receptors. The levorotatory isomer showed exceptionally high affinity for the CB1 receptor with a seven-fold selectivity over CB2. PMID- 12543519 TI - Mind and matter as asymptotically disjoint, inequivalent representations with broken time-reversal symmetry. AB - Many philosophical and scientific discussions of topics of mind-matter research make implicit assumptions, in various guises, about the distinction between mind and matter. Currently predominant positions are based on either reduction or emergence, providing either monistic or dualistic scenarios. A more-involved framework of thinking, which can be traced back to Spinoza and Leibniz, combines the two scenarios, dualistic (with mind and matter separated) and monistic (with mind and matter unseparated), in one single picture. Based on such a picture, the transition from a domain with mind and matter unseparated to separate mental and material domains can be viewed as a result of a general kind of symmetry breaking, which can be described formally in terms of inequivalent representations. The possibility of whether this symmetry breaking might be connected to the emergence of temporal directions from temporally non-directed or even non-temporal levels of reality will be discussed. Correlations between mental and material aspects of reality could then be imagined as remnants of such primordial levels. Different conceivable types of inequivalent representations would lead to correlations with different characteristics. PMID- 12543518 TI - Dynamics of nutrient-phytoplankton interaction in the presence of viral infection. AB - The present paper deals with the problem of a nutrient-phytoplankton (N-P) populations where phytoplankton population is divided into two groups, namely susceptible phytoplankton and infected phytoplankton. Conditions for coexistence or extinction of populations are derived taking into account general nutrient uptake functions and Holling type-II functional response as an example. It is observed that the three component systems persist when the infected phytoplankton population is not able to consume nutrient. PMID- 12543520 TI - Different facets of a computational equivalent of genetic addition. AB - This study deals with a biologically inspired system called DNA Addition System (DAS). The system consists of four elements: a chain molecule; a set of free (elemental) molecules, all of which refer to the elements of DNA; a quasi-free molecule, which is the last elemental molecule of the chain molecule; and a molecular machine. When the system works, a definite reconfiguration, referred to as addition, takes place. Addition actually adds a free molecule to the chain molecule to make it grow longer. The molecular machine decides which type of free molecule should be added taking into account the informational state of the system. A simulation model is also developed based on DAS to help generate chain molecule applying successive additions. This model can be used to simulate chain molecules that exhibit the characteristics of genetic molecules (i.e. molecules with identifiable segments) and behaviors of dynamical systems. How to correlate the characteristics of a dynamical system with the characteristics of DAS is also pointed out. Further study should be carried out to enhance the knowledge of DAS from both theory and application points of view. PMID- 12543521 TI - A search for conceptual congruence between individuals and species. AB - A new argument that species are individuals has recently been presented based on the way species names are used and systemic logic. The alternate idea that species are a categorical item, i.e., collections of individuals, was rejected. We wanted to evaluate this approach by establishing concepts that apply to individuals in order to examine if they are also applicable to a species viewed as a categorical item. It is our thinking that if species are not such, then concepts applicable to individuals will not be applicable to species so viewed. Individuals and species are similar in the presence of conceptual boundaries, unidirectional change and the presence of past and future time. These are all central, and salient, biological phenomena. But, these are concepts that apply also to populations which, for the most part, are not the same as species. Thus, these considerations infer that species, when viewed as a collection of individuals, do not share relevant concepts with individuals. Our conclusion is that a species is a 'unity' rather than an 'individual'. PMID- 12543522 TI - On the number of states of the neuronal sources. AB - In a previous paper (Proceedings of the World Congress on Neuroinformatics (2001)) the authors applied the so-called Lempel-Ziv complexity to study neural discharges (spike trains) from an information-theoretical point of view. Along with other results, it is shown there that this concept of complexity allows to characterize the responses of primary visual cortical neurons to both random and periodic stimuli. To this aim we modeled the neurons as information sources and the spike trains as messages generated by them. In this paper, we study further consequences of this mathematical approach, this time concerning the number of states of such neuronal information sources. In this context, the state of an information source means an internal degree of freedom (or parameter) which allows outputs with more general stochastic properties, since symbol generation probabilities at every time step may additionally depend on the value of the current state of the neuron. Furthermore, if the source is ergodic and Markovian, the number of states is directly related to the stochastic dependence lag of the source and provides a measure of the autocorrelation of its messages. Here, we find that the number of states of the neurons depends on the kind of stimulus and the type of preparation ( in vivo versus in vitro recordings), thus providing another way of differentiating neuronal responses. In particular, we observed that (for the encoding methods considered) in vitro sources have a higher lag than in vivo sources for periodic stimuli. This supports the conclusion put forward in the paper mentioned above that, for the same kind of stimulus, in vivo responses are more random (hence, more difficult to compress) than in vitro responses and, consequently, the former transmit more information than the latter. PMID- 12543523 TI - Field-induced reorganization of the neural membrane lipid bilayer: a proposed role in the regulation of ion-channel dynamics. AB - We present a computational model demonstrating that an electric field propagating in the plane of the neural membrane during transmembrane ion movement creates lateral concentration gradients of the lipids. Due to this field-induced reorganization, ethenes of the lipid chains become aligned and polarized. This finding is interpreted within the context of molecular studies of protein folding in biological membranes. We propose that electrostatic interactions between membrane dipoles and charged amino acid residues of the unfolded ion-channel protein regulate protein-folding kinetics (channel closing). These electrostatic interactions thus regulate electrical signaling in neurons. PMID- 12543524 TI - Evaluating health interventions in the 21st century: old and new challenges. PMID- 12543525 TI - The development of health technology assessment. AB - The field of health technology assessment (HTA) is still relatively new, but it has shown remarkable growth over the last decade, having spread first from the United States to Europe, and now to the entire world. HTA seeks to couple evidence with decision-making, and thus has similarities to evidence-based health care and evidence-based policy-making. The early history of HTA, beginning around 1975, reveals a first period of synthesising available evidence-principally that dealing with efficacy and cost-effectiveness of health care interventions-so as to put it in a format helpful to health policy-makers, especially those in national governments. From 1985 or so, the focus of the second period was on seeking more effective links with these policy-makers, particularly in Europe. The most recent period, beginning in the late 1990s, has been increasingly devoted to more effective dissemination and implementation in order to influence administrators and clinicians. While early assessments tended to focus on large, expensive, machine-based technologies, the scope has gradually widened to include smaller technologies, 'softer' technologies (such as counselling), and health care needs. Actual assessments have also taken on broader issues, such as organisational, social, and ethical implications. In the Member States of the European Union (EU), HTA activities are increasingly visible, and almost all now have a national focus for HTA associated with the Ministry of Health or its equivalent. Central and Eastern European countries are also developing HTA activities. Most recently, HTA has been highlighted by health policy documents from the European Commission. It seems likely that HTA will in the future be institutionalised in some form as part of EU activities. PMID- 12543526 TI - Evaluating health care interventions in the European Union. AB - This paper examines the current state of evaluations of health care interventions in the European Union, from the identification and commissioning of research through to its impact on policy and practice. Material is drawn from a survey conducted for the ASTEC project as well as a review of literature. Although the use of evaluative research has increased substantially in the last decade, both the pace of change and preferred research methodologies employed differ markedly. Much research still concentrates on issues of safety, efficacy and effectiveness, although there is evidence of an increasing emphasis on cost-effectiveness. Many countries are beginning to introduce systems linking economic evaluation to the decision-making process, while networks for the exchange of information continue to evolve. Research capacity in the public sector, although improving, is uneven, in part due to the uncertainty over long term career prospects and competition from industry. Capacity building measures should in particular ensure that dissemination expertise is strengthened, and that more emphasis is placed on developing receptor capacity within different stakeholder groups. Linking knowledge production to changes in practice remains a key challenge. Further research on implementation and impact assessment is required, to help demonstrate the value of evaluations on both policy and practice. PMID- 12543527 TI - Setting priorities for the evaluation of health interventions: when theory does not meet practice. AB - Priority setting is a key component of the process of evaluating health interventions. This has traditionally been an informal process led by power and influence, but a number of explicit criteria and systematic models have been developed since the late 1980s. This paper presents a review and appraisal of these conceptual models and examines how they have influenced the practice of priority setting in the United States and Europe. The main conclusion is that a significant gap exists between theory and practice. Most models have been developed for the purpose of maximising health gains through an efficient allocation of resources. However, they present at least three important limitations that need to be removed if formal models are to play a more substantial role in decision making: they tend to prioritize interventions for evaluation, rather than evaluations themselves; they fail to address priority setting in a research portfolio perspective; and they fail to adopt an incremental perspective. Existing prioritization models are not suitable for supporting cost-containment or distributional objectives. PMID- 12543528 TI - From health technology assessment to research on the organisation and delivery of health services: addressing the balance. AB - This paper argues that the focus of research to improve health services has, until recently, been on health technology assessment. The authors make the case for a greater emphasis on research on how health services are managed, organised and delivered, and refer to initiatives in a number of countries which are seeking to address this balance. The way two such initiatives in England and Canada have set priorities for this type of research, involving a wide range of stakeholders is described. The authors argue that a wide range of disciplines needs to be applied to research on the organisation and delivery of health services. Important theoretical differences between and within disciplines, and their implications for research methods, are discussed. An example of an issue in the delivery of organisation of health services (how best to deliver orthopaedic care) is used to illustrate how a number of different disciplines can be applied. The challenge for researchers from these disciplines is to see how far they can work together to carry out research in this important field. The challenge for this research is that the findings are valued and used by health service professionals, managers and users. PMID- 12543529 TI - Regulating the economic evaluation of pharmaceuticals and medical devices: a European perspective. AB - Throughout the developed world, economic evaluation of costly new pharmaceuticals and medical devices became increasingly widespread and systematic during the 1990s. However, serious concerns remain about the validity and relevance of this economic evidence, and about the transparency and accountability of its use in public sector reimbursement decisions. In this article, we summarise current concerns in Europe, based on interviews with European health economists from industry, universities, research institutes and consulting firms. We identify five challenges for European policy-makers, and conclude that there is considerable scope for improving decision-making without damaging incentives to innovate. The challenges are: (1). full publication of the economic evidence used in reimbursement decisions; (2). the redesign of licensing laws to improve the relevance of economic data available at product launch; (3). harmonisation of economic evaluation methodologies; (4). development of methodologies for evaluation of health inequality impacts; and (5). negotiation of price performance deals to facilitate the use of economic evidence in post-launch pricing review decisions, as information is gathered from studies of product performance in routine use. PMID- 12543530 TI - The role of the media in steering public opinion on healthcare issues. AB - The media can play an important role in influencing both the demand and supply of medical treatments, regardless of evidence of effectiveness. This can be highlighted by recent experiences in Italy following publicity for a new unproven treatment in Italy, which it was claimed was highly successful in treating a wide range of cancers. The media role in influencing both health authorities to fund large scale trials of the intervention and patients awareness and expectations of therapy are discussed. The changes in Italian media over the last 20 years have seen a reduction in the numbers of specialist medical journals, with increased emphasis placed on sensationalism rather than accuracy. The media though has the potential to play in future an important strategic role in disseminating accurate information on issues pertaining to health. PMID- 12543531 TI - Evaluations of health interventions in social insurance-based countries: Germany, the Netherlands, and Austria. AB - Health Technology Assessment (HTA) in social insurance-based, or so-called 'Bismarck' health care systems (Germany, Austria, and the Netherlands) has taken a different course than in either taxed-based (Sweden, Norway, United Kingdom, and Spain) or private health care systems (such as the United States). The culture of informed decisions supported by transparent and evidence-based evaluations of health interventions was hindered by the strong professional autonomy and sectoral interests in Germany and Austria for a long time. On the other hand, HTA has a long-standing tradition in the Netherlands. In all three countries sickness funds play an important role in implementing evaluations-as a policy tool-by linking reimbursement to explicit proof of effectiveness in both new and established interventions. This article focuses on the obstacles and opportunities for HTA in Germany, Austria and the Netherlands as countries with insurance-based health care systems. PMID- 12543532 TI - Health economic evaluation in Japan: a case study of one aspect of health technology assessment. AB - There is a burgeoning literature in health economic evaluation, with this form of analysis becoming increasingly influential at the health policy making level in a number of countries. However, a search of the literature reveals that in Japan, the world's second largest health care market, very little health economic evaluation has been undertaken. The main reason for the lack of interest in economic evaluation is that the fee-for-service and strict price regulation that characterises the system of health care financing in Japan is not conducive to this form of analysis. Moreover, the government and many researchers are satisfied that the current organisation of health care has given long life and low infant mortality at low cost. Even if it is accepted that low health care costs and good health prevail in Japan, slower economic growth rates, an ageing population and the development of new medical technologies will place increasing pressure on health care resources and will necessitate a more rational use of these resources. Good economic evaluation, by weighing benefits against costs, has an important role to play. PMID- 12543533 TI - Co-ordinating health technology assessment in Canada: a European perspective. AB - Health Technology Assessment (HTA) is well-established in Canada, with both a national co-ordinating office for HTA and several provincial HTA organisations. The success of and relationship between these agencies, may provide useful lessons for policy-makers considering how to facilitate increased co-operation and harmonisation of HTA activity across Europe. Interviews were conducted with a number of key individuals associated with HTA in Canada to identify achievements, examine the relationship between the provincial agencies and the national co ordinating office, and detect barriers to the increased use of HTA knowledge in decision-making. PMID- 12543534 TI - Evaluating health interventions: exploiting the potential. AB - The importance of the evaluation of health care interventions (EHI) including formal health technology assessment (HTA) cannot be over-emphasised, as its results can inform and improve resource allocation decisions in all parts of the health care system, public and private. At present, fragmented and inefficient resource allocation processes are a universal problem and, as a consequence, patients are deprived of care from which they could benefit. Such outcomes are not only inefficient but also unethical. In this paper we focus on HTA, which emphasises the need to link evidence to policy and practice, but our conclusions can be applied to all types of EHI. There is a need for greater use of economic evidence within HTA. Four principle characteristics of HTA are discussed in this paper: types of knowledge and evaluation, and commissioning and knowledge utilisation. Policy-makers must invest in improving research and receptor capacities for producing and using knowledge, improve quality control, and invest in research into dissemination and implementation mechanisms. The potential of HTA is great, and its use has increased, but it remains largely unexploited in most countries. PMID- 12543535 TI - Biofilm testing of Staphylococcus epidermidis clinical isolates: low performance of vancomycin in relation to other antibiotics. AB - The in vitro killing effect of widely used antibiotics (cephalothin, clindamycin, erythromycin, ofloxacin, rifampicin, teicoplanin, tetracycline, phosphomycin and vancomycin) was comparatively analyzed in this study on 24-h biofilms of 64 Staphylococcus epidermidis clinical isolates. This effect was assessed at the expected antibiotic concentration reached in serum, using ATP-bioluminescence. Erythromycin, rifampicin, tetracycline and phosphomycin presented generally a higher killing effect than vancomycin, clindamycin, cephalothin, teicoplanin and ofloxacin in these biofilms. Differences in the resistance profiles obtained in classical assays (broth microdilution and diffusion) did not help to predict differences in the killing effect of the antibiotics in biofilms. Only some antibiotics (vancomycin but not rifampicin or tetracycline) highly decreased their killing effect as the biofilm age increased (from 6 to 24 or 48 h). These studies underline the relevance of biofilm susceptibility testing and the potential danger of the indiscriminate use of vancomycin monotherapy as the ultimate resource against infections involving aged biofilms. PMID- 12543536 TI - Epidemiology and outcome of Clostridium difficile infection and diarrhea in HIV infected inpatients. AB - Clostridium difficile causes diarrhea in HIV infected patients but reports of prevalence, risk factors, and outcome vary. We studied the impact of C. difficile in 161 HIV infected inpatients admitted to Cook County Hospital. Patients with C. difficile had more hospital admissions in the previous 6 months (p =.04), spent more days in the hospital in the previous 3 months (p =.02), more often had previously received H2 blockers or treatment for Pneumocystis carinii (p <.05), and had a more frequent history of herpesvirus (p =.03) or opportunistic infections (p =.04). C. difficile associated diarrhea (CDAD) was the etiology in 32% of all study patients with diarrhea. Patients with CDAD were hospitalized for longer periods (p =.02) and received more antibiotics (p =.002). C. difficile was frequently present in our HIV infected patients, especially those with advanced HIV disease, but appeared to have little impact on morbidity or mortality. PMID- 12543537 TI - Prevalent transfer of human colostral IgA antibody activity for the enteropathogenic Escherichia coli bundle-forming pilus structural repeating subunit A in neonates. AB - The frequency of IgA antibody activity to the structural protein subunit BfpA of the enteropathogenic Escherichia coli bundle-forming pilus was determined in 40 mother-infant pairs by immunoblot analysis using affinity purified recombinant BfpA to monitor for IgA in maternal colostrum and in feces of the neonates. Fecal samples were collected from exclusively breastfed term infants < 24-h after the first breastmilk feeding and colostral samples from their mothers. Infants were monitored prospectively with monthly visits to ascertain dietary practices and diarrheal illnesses. The percentage of colostral anti-BfpA IgA positive patients that were also coproantibody positive was 67.5%. The median duration of lactation was 108 days and the incidence of infantile diarrheal disease was 7.5%. Thus, colostral anti-BfpA IgA antibody activity survives passage through the gut of breastfed neonates, persisting in their feces. It is suggested that oral passive immunotherapy may be used to prevent and/or treat typical EPEC infection during infancy. PMID- 12543538 TI - Characteristics and association with disease of two major subclones of Shiga toxin (Verocytotoxin)-producing strains of Escherichia coli (STEC) O157 that are present among isolates from patients in Germany. AB - Shiga toxin (Verocytotoxin) producing E. coli (STEC) O157 were isolated from 168 patients living in different parts of Germany. Most isolates were from sporadic cases and seven small outbreaks with STEC O157 were identified. The 168 strains were examined for phenotypic and genotypical traits in order to identify major types of STEC O157 occurring in Germany. Phage typing (PT) revealed PT8 (n = 54) and PT2 (n = 48) strains as most frequent (60.7%) among the isolates. Carriage of the stx(2) gene by STEC O157 was closely associated with hemolytic uremic syndrome (100%) and with bloody diarrhea (61.7%). The stx(2) gene was frequent in PT88, PT47 (both 100%), PT2 (91.5%) and PT4 (87.5%) strains and more rarely (33.3%) found in strains belonging to the other PTs. PT8 and PT2 strains formed two groups which differed from each other in their motility, stx-genotypes and the severity of the illness they caused. Pulsed-field gel electrophoresis of PT2 and PT8 strains and hybridization of XbaI digested DNA with stx(1) and stx(2) specific gene probes revealed similarities among epidemiologically unrelated strains belonging to the same PT. The results indicate that STEC O157 PT2 and PT8 strains form two distinct subclones which are dominating in Germany and other European countries. PMID- 12543539 TI - Evaluation of BACTEC MGIT 960 PZA medium for susceptibility testing of Mycobacterium tuberculosis to pyrazinamide (PZA): compared with the results of pyrazinamidase assay and Kyokuto PZA test. AB - The fully automated BACTEC MGIT 960 PZA medium for susceptibility testing of Mycobacterium tuberculosis to pyrazinamide (PZA) was evaluated using 101 Mycobacterium tuberculosis clinical isolates. The results obtained with the system were compared with those of the pyrazinamidase (PZase) assay and the Kyokuto PZA test based on a broth culture, which is commercially available in Japan. The overall concordance rate was 90.1% (91/101) among the three methods in the initial test. The concordance rates between the BACTEC MGIT 960 PZA medium vs the PZase assay, the BACTEC MGIT 960 PZA medium vs the Kyokuto PZA test, and the PZase assay vs the Kyokuto PZA test were 93.1, 91.1, and 96.0%, respectively. On the repeat test of the 10 strains with discrepant results among the three methods, the concordance rates reached over 97% between each of the two systems. The results of the repeat test were confirmed by MIC testing and sequencing analysis of the pncA gene encoding PZase of M. tuberculosis. The mean turnaround times from incubation for PZA susceptibility testing were almost similar for the two methods based on liquid media, the BACTEC MGIT 960 PZA medium and the Kyokuto PZA test (7.7 and 7.4 days, respectively). These results indicate that both methods based on liquid media, the fully automated BACTEC MGIT 960 PZA medium and the Kyokuto PZA test for susceptibility testing to PZA, are useful for rapid diagnosis of PZA resistant tuberculosis. PMID- 12543540 TI - Polymicrobial candidemia. AB - We conducted a retrospective review of the clinical data on patients with polymicrobial candidemia during a nine-year period (1992-2000) at our tertiary care University Hospital. Also, the clinical features of patients with polymicrobial vs monomicrobial candidemia were compared. There were 16 (5.2%) patients with multiple candidal species in blood among 303 patients with candidemia. Patients' age varied from 21-85 years (median 52 years); they had serious underlying co-morbidities with prolonged hospitalization (median duration 13 days); all had intravenous vascular catheters, had been exposed to multiple antibiotics and were heavily colonized with Candida. Concomitant serious bacterial infections were common (56%). Candida albicans was isolated from blood in 11 of 16 patients; other species were C. glabrata (7 patients), C. tropicalis (6 patients) and C. parapsilosis (4 patients). Sixty seven percent (8 of 12) patients had causative Candida species isolated from vascular catheter tip culture. Polymicrobial candidemia occurred in sicker, non-oncologic patients with frequent concomitant bacterial infections, as compared to those with monomicrobial candidemia. Crude mortality was 43%, a rate similar to that seen with monomicrobial candidemia. Polymicrobial candidemia is uncommon, seen in hospitalized patients with multiple co-morbidities and heavy candidal colonization; removal of vascular catheter and institution of antifungal therapy are important therapeutic maneuvers. PMID- 12543541 TI - In vitro susceptibility of Mycoplasma hominis clinical isolates to tetracyclines, quinolones and macrolides. AB - We tested the in vitro activity of levofloxacin, ciprofloxacin, doxycycline, tetracycline, erythromycin, roxithromycin, clarithromycin and azithromycin against 110 clinical isolates of Mycoplasma hominis. The minimal inhibitory concentrations (MICs) were determined with the Etest. The minimal concentrations at which 90% of the isolates were inhibited (MIC(90)) were 0.064 microg/ml doxycycline and 0.19 microg/ml tetracycline. In 9 isolates (8.1%), the MIC for doxycycline was 4-12 microg/ml. These isolates were also resistant to tetracycline with a MIC of 32-128 microg/ml. No significant difference was found between doxycycline and tetracycline (p = 0.076). Comparison of the two quinolones revealed that the MIC(90) for levofloxacin was 0.19 microg/ml and for ciprofloxacin, 0.5 microg/ml. A significant difference was found between doxycycline/tetracycline and levofloxacin or ciprofloxacin (p = 0.0001), and between levofloxacin and ciprofloxacin (p = 0.001). All the isolates were highly resistant to the macrolides with MIC > or = 256 microg/ml. This finding has important implications for cases in which Mycoplasma infection is suspected and culture and/or in vitro susceptibility tests are not available. PMID- 12543542 TI - Comparison of methods of interpretation of checkerboard synergy testing. AB - Four different methods for interpreting the results of checkerboard synergy testing were compared by applying each to a set of synergy study data. Statistically significant differences in synergy were detected among methods (% synergy ranged from 10 to 83%). As interpretations were found to vary widely based upon method, one should be aware of this in interpreting the relevant literature. PMID- 12543544 TI - Antimicrobial activity of broad-spectrum agents tested against Gram-negative bacilli resistant to ceftazidime: report from the SENTRY Antimicrobial Surveillance Program (North America, 2001). AB - To assess the effect of ceftazidime resistance on the activity of other antimicrobial agents, 4,175 Gram-negative bacilli isolates, designated as ceftazidime-susceptible (3,759, 90%) or -resistant (416, 10%) were tested against a panel of nine agents. Cefepime was the most active of the agents tested, against both ceftazidime-susceptible and -resistant isolates, with a particularly low incidence (0.3%, 10/3152 isolates) of resistance among the Enterobacteriaceae. Tobramycin had the best activity against Pseudomonas aeruginosa and ticarcillin/clavulanate was the least active of the tested agents. This study confirms the need for constant monitoring of in vitro antimicrobial efficacy to direct empiric treatment regimens. PMID- 12543543 TI - Evaluation of the in vitro activity of six broad-spectrum beta-lactam antimicrobial agents tested against recent clinical isolates from India: a survey of ten medical center laboratories. AB - The widespread use of beta-lactam antimicrobial agents as first-line therapy for the treatment of serious infections has led to the development of various resistances that have compromised the use of some agents. In certain countries, the lack of local or national surveillance programs limits the ability to detect these resistant strains and prevent their dissemination. A 10 medical center study in India was initiated to benchmark prevailing resistance rates for a range of bacterial pathogens to beta-lactams, and it found high rates of beta-lactamase mediated resistance in Escherichia coli and Klebsiella spp. These rates included: cephalosporins (55.6-61.3% resistance), with extended-spectrum beta-lactamase (ESBL) phenotypes noted in over 60% of E. coli isolates and in Salmonella spp. (3.2-8.1%). Imipenem, a carbapenem, was the only antimicrobial agent tested with 100% activity against Enterobacteriaceae. Cefpirome was the most active of the tested cephalosporins, and all were fully active against methicillin-susceptible staphylococci with the exception of ceftazidime. Molecular and susceptibility characterization of 52 selected ESBL-producing strains showed a high level of co resistance with aminoglycosides and fluoroquinolones, and clonal dissemination of resistant strains within medical centers. Collaborative studies, such as those presented here, can accurately detect changes in resistance patterns, and their continued use may help limit the further development and spread of bacterial resistances in India. PMID- 12543545 TI - Molecular identification of Paracoccidioides brasiliensis by 5' nuclease assay. AB - A PCR assay based on the 5' nuclease assay using a fluorescent probe derived from the sequence of the gene coding for the 43,000-Da (gp43) antigen was developed to detect Paracoccidioides brasiliensis. The assay could detect at least 10 copies of this DNA sequence, providing efficient accuracy to be useful for diagnosis of paracoccidioidomycosis. PMID- 12543546 TI - Biological activity of immunostimulatory CpG DNA motifs in domestic animals. AB - Bacterial DNA contains a much higher frequency of CpG dinucleotides than are present in mammalian DNA. Furthermore, bacterial CpG dinucleotides are often not methylated. It is thought that these two features in combination with specific flanking bases constitute a CpG motif that is recognized as a "danger" signal by the innate immune system of mammals and therefore an immune response is induced when these motifs are encountered. These immunostimulatory activities of bacterial CpG DNA can also be achieved with synthetic CpG oligodeoxynucleotides (ODN). Recognition of CpG motifs by the innate immune system requires engagement of Toll-like receptor 9 (TLR-9), which induces cell signaling and subsequently triggers a pro-inflammatory cytokine response and a predominantly Th1-type immune response. CpG ODN-induced innate and adaptive immune responses can result in protection in various mouse models of disease. Based on these observations, clinical trials are currently underway in humans to evaluate CpG ODN therapies for cancer, allergy and infectious disease. However, potential applications for immunostimulatory CpG ODN in species of veterinary importance are just being explored. In this review, we will highlight what is presently known about the immunostimulatory effects of CpG ODN in domestic animals. PMID- 12543547 TI - Cutaneous hypersensitivity reactions to Psoroptes ovis and Der p 1 in sheep previously infested with P. ovis--the sheep scab mite. AB - We have previously shown that infestation with Psoroptes ovis induces an IgE response and intense tissue eosinophilia, typical of a Type I hypersensitivity response [Parasite Immunol. 22 (2000) 407]. Intradermal tests (IDSTs) suggest that there are also delayed and Arthus-type responses to this parasite. In order to study the nature of ovine cutaneous reactions to P. ovis, naive controls and experimentally infested sheep (n = 5) were challenged intradermally with mite antigen. Challenge elicited immediate (P < 0.001) and delayed (P < 0.005) wheal reactions in sensitised sheep. At 6 (P < 0.02) and 30 h (P < 0.001) the predominant infiltrating cells were eosinophils. To explore the role of circulating antibodies, naive sheep (n = 5) were subjected to Prausnitz-Kustner (PK) tests. These elicited immediate (P < 0.02) but not delayed wheal reactions. At 6 h eosinophils (P < 0.001) dominated the infiltrate. These results suggest that P. ovis allergens provoke an IgE-dependent immediate and late phase response and a cell-mediated eosinophil-rich delayed-type hypersensitivity response (ER DTH). PMID- 12543548 TI - Equine platelet CD62P (P-selectin) expression: a phenotypic and morphologic study. AB - Acute inflammatory diseases, such as colic, septicemia and endotoxemia are common in equines and have been shown to be correlated to vascular injury and thrombosis. In humans with similar thrombotic conditions, P-selectin and P selectin glycoprotein ligand-1 (PSGL-1)-mediated platelet-leukocyte adhesion contributes to the pathogenesis of these disorders through the generation of inflammatory mediators and tissue factor. As such, we hypothesized that a P selectin-PSGL-1 (platelet-leukocyte) interaction, similar to that in humans, may also exist in the horse. The objective of this study was to investigate phenotypic and morphological properties of equine platelet activation with a focus on CD62P (P-selectin) expression and CD62P mediated platelet-leukocyte interactions. To study high levels of platelet activation, we used 1 U/ml thrombin to induce secondary, irreversible aggregation in both human and equine platelets. Addition of glycyl-L-prolyl-L-arginyl-L-proline amide (GPRP) prior to thrombin activation blocked fibrin polymerization, allowing the use of flow cytometry to study alpha-granule expression as a measure of platelet activation. Thrombin activation resulted in high levels of activation, measured as P-selectin expression, in both humans and equines. Interestingly, our research illustrates that in healthy horses, P-selectin is also constitutively expressed on 20-25% of resting platelets. This finding is in direct contrast to humans, in which P selectin expression is negligible (<5%) in the absence of agonist activation. The high baseline level of P-selectin expression among equine platelets may suggest that they are primed for leukocyte adhesion, possibly resulting in prothrombotic conditions. This phenomenon could be of significant clinical relevance, as it may be related to the rapid clinical decline often seen in equine patients with colic and endotoxemia, where vascular injury and thrombotic complications compromise patient survival. Based on these findings, further investigation into the mechanisms of platelet P-selectin-mediated inflammation and platelet-leukocyte mediated vascular injury in the horse appears warranted. PMID- 12543549 TI - Immunomodulatory effects of alpha melanocyte stimulating hormone on common carp (Cyprinus carpio L.). AB - The immunomodulatory effects of phagocytic cells in common carp Cyprinus carpio L., by alpha melanocyte stimulating hormone (alpha-MSH) was analyzed in vitro. Carp head kidney leucocytes were cultured in RPMI 1640 medium containing 0.05, 0.1, 1 and 10 nM alpha-MSH and the production of superoxide anion was measured via the reduction of nitroblue tetrazolium (NBT) in vitro. Macrophages incubated with alpha-MSH showed an increase in the production of superoxide anion in comparison to control macrophages cultured without hormone. Phagocytic cells treated with alpha-MSH also displayed increased phagocytosis. Furthermore, carp lymphocytes treated with alpha-MSH increased the mitogenic responses to phytohaemoagglutinin (PHA-P). These results show that alpha-MSH directly influences fish immune responses. PMID- 12543550 TI - Control of immunoglobulin isotype production by porcine B-cells cultured with cytokines. AB - Cytokines regulate immunoglobulin (Ig) isotype production following the Th1/Th2 paradigm, derived from studies of inbred mice. In pigs, it is not known which, if any, Ig isotypes may reflect a Th1/Th2 response. To evaluate this, purified porcine CD21(+) B-cells were co-cultured with Staphylococcus aureus Cowan strain 1 or Escherichia coli lipopolysaccharide as B-cell mitogens together with recombinant human IL-2, and recombinant porcine (rp) interferon (IFN)-gamma, IL 12 or IL-10. While the mitogens increased B-cell proliferation, cytokines had no additional effect. A quantitative competitive enzyme-immuno assay was used to measure concentrations of porcine IgM, IgG(1) and IgG(2) in B-cell culture supernatants. In vitro, porcine B-cells produced IgG(2), 106 +/- 17.3 microg/ml; IgG(1) 107 +/- 38.3 microg/ml and IgM 25.6 +/- 8.45 microg/ml. In some individuals, Th1 cytokines such as rpIFN-gamma and IL-12, enhanced IgG(2) in the face of low concentrations of IgG(1). Furthermore, individual responses, in some cases, tended to be diametrically opposed, reminiscent of previously documented categorical immune responses in pigs such that some individuals produced high concentrations of IgG(1) in response to the various doses of rp cytokines, while others produced lower concentrations. Pigs may generate a high IgG(1):IgG(2) ratio in response to rpIL-10, and possibly to other Th2-associated cytokines. However, B-cell response to rp cytokines in vitro exhibits marked variation by pig, a feature that is likely a function of highly variable individual genotypes and their interaction with complex environments. PMID- 12543551 TI - Cloning of porcine interleukin (IL)-12 receptor beta2 (IL-12Rbeta2) gene and its application to a rapid biological assay for human/porcine IL-12. AB - Porcine IL-12Rbeta2 gene was cloned from mRNA preparation of mitogen-activated peripheral blood mononuclear cells (PBMCs), and its complete nucleotide sequence was determined. To confirm the biological function, the entire open reading frame (ORF) was re-cloned into a mammalian expression vector, pcDNA3.1/Zeo(+), at the downstream of CMV promoter, and introduced to a Th1-like human lymphoma cell line, Jurkat E6-1. Antibiotic-resistant cells retaining the expression construct were selected then, isolated by the limiting dilution method. An established clone (10B10) constitutively expressed chimeric IL-12Rs composed of intrinsic (human) beta1 and extrinsic (porcine) beta2 subunits, and produced interferon (IFN)-gamma in response to IL-12 of both species with optimal PHA/PMA stimulation. The production of IFN-gamma was observed as early as 42 h after culture and appeared to be dose-dependent within the range between 20 and 2000 pg/ml. Thus, this clone not only reacts with IL-12 of both species but also provides a useful tool for quick and sensitive detection of IL-12 bioactivity. PMID- 12543553 TI - Synthesis and potential antimetastatic activity of monovalent and divalent beta-D galactopyranosyl-(1-->4)-2-acetamido-2-deoxy-D-glucopyranosides. AB - Anomers of monovalent and divalent beta-D-galactopyranosyl-(1-->4)-2-acetamido-2 deoxy-D-gluco-pyranosides were synthesized under different glycosylation conditions, and evaluated for in vitro antimetastatic activity. Three compounds showed promising inhibitory effects on cancer cell attachment, spreading, migration, and invasion. PMID- 12543554 TI - Purification and characterization of an endo-beta-(1-->6)-galactanase from Trichoderma viride. AB - An endo-beta-(1-->6)-galactanase from Onozuka R-10, a commercial cellulase preparation from Trichoderma viride, was purified 57-fold. Apparent Mr values of the purified enzyme, estimated by denaturing gel electrophoresis and gel filtration, were 47,000 and 17,000, respectively. The enzyme was assayed with a galactan from Prototheca zopfii, which has a high proportion of beta-(1-->6) linked galactosyl residues. It exhibited maximal activity toward the galactan at pH 4.3. The enzyme hydrolyzed specifically beta-(1-->6)-galactooligosaccharides with a degree of polymerization higher than 3 and their acidic derivatives with 4 O-methyl-glucosyluronic or glucosyluronic groups at the nonreducing terminals. The methyl beta-glycoside of beta-(1-->6)-galactohexaose was degraded to reducing galactooligomers with a degree of polymerization 2-5 as the products at the initial stage of hydrolysis, and galactose and galactobiose at the final stage, indicating that the enzyme can be classified as an endo-galactanase. The extent of hydrolysis of the carbohydrate portion of a radish root arabinogalactan protein (AGP) increased when alpha-L-arabinofuranosyl residues attached to beta (1-->6)-linked galactosyl side chains of the AGP were removed in advance. The enzyme released galactose, beta-(1-->6)-galactobiose, and 4-O-methyl-beta glucuronosyl-(1-->6)-galactose as major hydrolysis products when allowed to act exhaustively on the modified AGP. PMID- 12543555 TI - Epoxidation of C-branched glycals: unexpected stereochemical results and their theoretical rationale. AB - This paper describes the synthesis of C-3 methyl-branched glycosides by epoxidation of partially unblocked L-configured glycals. The stereochemical result depends on the orientation of the allylic hydroxyl group. A theoretical explanation is presented, based on the conformational preferences of the respective glycal half-chair conformations that were estimated by applying the BP density functional and a valence triple-zeta basis set. PMID- 12543556 TI - Structural determination of the lipo-chitin oligosaccharide nodulation signals produced by Rhizobium giardinii bv. giardinii H152. AB - Rhizobium giardinii bv. giardinii is a microsymbiont of plants of the genus Phaseolus and produces extracellular signal molecules that are able to induce deformation of root hairs and nodule organogenesis. We report here the structures of seven lipochitooligosaccharide (LCO) signal molecules secreted by R. giardinii bv. giardinii H152. Six of them are pentamers of GlcNAc carrying C 16:0, C 18:0, C 20:0 and C 18:1 fatty acyl chains on the non-reducing terminal residue. Four are sulfated at C-6 of the reducing terminal residue and one is acetylated in the same position. Six of them are N-methylated on the non-reducing GlcN residue and all the nodulation factors are carbamoylated on C-6 of the non-reducing terminal residue. The structures were determined using monosaccharide composition and methylation analyses, 1D- and 2D-NMR experiments and a range of mass spectrometric techniques. The position of the carbamoyl substituent on the non reducing glucosamine residue was determined using a CID-MSMS experiment and an HMBC experiment. PMID- 12543557 TI - Heparin antiproliferative activity on bovine pulmonary artery smooth muscle cells requires both N-acetylation and N-sulfonation. AB - The antiproliferative activity of Heparin (HP) on bovine pulmonary artery smooth muscle cells (BPASMC) in vitro requires both N-acetylation and N-sulfonation. This was demonstrated by quantifying the relative N-acetylation of three commercial heparins of known antiproliferative activities, using their Fourier transform infrared (FTIR) band areas at 1381-1378 and 1320-1317 cm(-1), which combined resulted in 1.0, 1.0 and 1.3 cm2 for Choay, Elkins-Sinn and Upjohn HP, respectively. These results show that Upjohn HP, which is at least 44% more antiproliferative than the other two, is 30% more N-acetylated. Upjohn HP was also N-desulfonated chemically, and its antiproliferative activity was determined. Its total sulfonate (--SO 3 -) content (O- and N-sulfonate) was quantified using the FTIR band area at 1260-1200 cm(-1) for the S=O stretching; a drop in sulfonate content from 21.87% (w/w) before N-desulfonation to 16.51% (w/w) after N-desulfonation, resulted in a 67% decrease in its inhibitory potency. In addition to the requirement that approximately 24% of the sulfonate content be bonded to N, the data show a direct correlation between the extent of Upjohn HP N-acetylation and its antiproliferative activity on BPASMC. PMID- 12543558 TI - Stereoselective synthesis of 1,2:4,5-di-O-isopropylidene-3-C-(5-phenyl-1,2,4 oxadiazol-3-yl)-beta-D-psicopyranose and its X-ray crystallographic analysis. AB - In a novel procedure, when 3-O-benzoyl-3-C-(N-hydroxycarbamimidoyl)-1,2:4,5-di-O isopropylidene-beta-D-psicopyranose (1) is treated with acetic anhydride, chloroacetyl chloride, propanic anhydride and benzoyl chloride, the 3-O-benzoyl group undergoes an intramolecular replacement reaction with neighbouring group participation and transfer resulting in a more stable conjugated system by the formation of a 1,2,4-oxadiazol ring. A possible mechanism is reported. The structure has been determined by spectroscopic data and X-ray crystallographic analysis. PMID- 12543559 TI - Purification and characterization of dermatan sulfate from the skin of the eel, Anguilla japonica. AB - Glycosaminoglycans were isolated from the eel skin (Anguilla japonica) by actinase and endonuclease digestions, followed by a beta-elimination reaction and DEAE-Sephacel chromatography. Dermatan sulfate was the major glycosaminoglycan in the eel skin with 88% of the total uronic acid. The content of the IdoA2Salpha1- >4GalNAc4S sequence in eel skin, which shows anticoagulant activity through binding to heparin cofactor II, was two times higher than that of dermatan sulfate from porcine skin. The anti-IIa activity of eel skin dermatan sulfate was determined to be 2.4 units/mg, whereas dermatan sulfate from porcine skin shows 23.2 units/mg. The average molecular weight of dermatan sulfate was determined by gel chromatography on a TSKgel G3000SWXL column as 14 kDa. Based on 1H NMR spectroscopy, the presence of 3-sulfated and/or 2,3-sulfated IdoA residues was suggested. The reason why highly sulfated dermatan sulfate does not show anticoagulant activity is discussed. In addition to dermatan sulfate, the eel skin contained a small amount of keratan sulfate, which was identified by keratanase treatment. PMID- 12543560 TI - N-(2-Carboxyethyl)chitosans: regioselective synthesis, characterisation and protolytic equilibria. AB - N-(2-Carboxyethyl)chitosans were obtained by reaction of low molecular weight chitosan with a low degree of acetylation and 3-halopropionic acids under mild alkaline media (pH 8-9, NaHCO3) at 60 degrees C. The chemical structure of the derivatives obtained was determined by 1H and 13C NMR spectroscopies. It was found that alkylation of chitosan by 3-halopropionic acids proceeds exclusively at the amino groups. The products obtained are described in terms of their degrees of carboxyethylation and ratio of mono-, di-substitution and free amine content. The protonation constants of amino and carboxylate groups of a series of N-(2-carboxyethyl)chitosans were determined by pH-titration at ionic strength 0.1 M KNO3 and 25 degrees C. PMID- 12543561 TI - Structural elucidation of the O-chain of the lipopolysaccharide from Xanthomonas campestris strain 8004. AB - A novel O-specific polysaccharide containing 3-acetamido-3-deoxy-alpha-D-fucose (Fuc3NAc) and D-rhamnose was isolated from the phenol-soluble lipopolysaccharide fraction of the plant associated bacterium Xanthomonas campestris strain 8004. The structure, determined by means of chemical analysis and 1D and 2D NMR spectroscopy, showed a branched trisaccharide repeating unit, as shown below: [formula: see text]. PMID- 12543562 TI - Synthesis and structural analyses of 3-acetamido-1,4-di-O-acetyl-2,3,5-trideoxy-5 C-(isopropylphosphinyl)-D-erythro-pentopyranoses. AB - 1H NMR spectroscopy of phosphorus containing hetero sugars (phospha sugars), revealed the alpha and beta configurations and chair conformations for 3 acetamido-1,4-di-O-acetyl-2,3,5-trideoxy-5-C-(isopropylphosphinyl)-alpha- and beta-D-erythro-pentopyranoses. The conformation of the title compounds was determined by 1H NMR as 1C4 in CDCl3 and the conformation was in accord with that in solid state determined by X-ray crystallographic analysis. PMID- 12543563 TI - Synthesis of anellated carbasugars from (--)-quinic acid. AB - (3R,4R,5R)-3-[(tert-Butyl-dimethylsilyl)oxy]-4,5-(isopropylidenedioxy)-1 cyclohexanone (2) reacted with carbon disulfide and methyl iodide in the presence of sodium hydride to furnish (3R,4R,5R)-5-[(tert-butyl-dimethylsilyl)oxy]-3,4 (isopropylidenedioxy)-2-[bis(methylthio)methylene]-1-cyclohexanone (3). 2 and N,N dimethylformamide dimethyl acetal afforded (2E,3R,4R,5R)-5-[(tert-butyl dimethylsilyl)oxy]-2-(dimethylaminomethylene)-3,4-(isopropylidenedioxy)-1 cyclohexanone (4). These push-pull activated methylenecyclohexanones 3 and 4 underwent a ring closure reaction with hydrazine hydrate and methylhydrazine, respectively, to give pyrazoloanellated carbasugars. Treatment of 3 with formamidinium, acetamidinium and benzamidinium salts, respectively, in the presence of sodium methanolate yielded three (5R,6R,7R)-7-[(tert-butyl dimethylsilyl)oxy]-5,6,7,8-tetrahydro-5,6 (isopropylidenedioxy)benzo[d]pyrimidines. PMID- 12543564 TI - Is aortic angiography necessary for accurate planning of endovascular aortic aneurysm stents? AB - Endovascular management of abdominal aortic aneurysms (AAA) relies upon accurate stent design. We hypothesized the length of the stent could be measured from computed tomogram (CT) generated 3D surface rendered images of the abdominal vessels, eliminating the need for digital subtraction angiography (DSA). METHODS: The CT scans from a consecutive series of 54 patients considered for endovascular AAA repair were reviewed. A TRIFAB Zenith (Cook) endovascular aortic stent was designed, using the manufacturer's sizing computer program. These hypothetical stents were compared to those that had been implanted, using measurements from both angiogram and CT (Spearman's test of correlation (p<0.05)). RESULTS: The stent body size and short limb extension (SLE) of stents in each group was similar (correlation coefficient 0.70(p<0.01) and 0.50 (p=0.014)), however, CT alone significantly underestimated the length of the long limb extension (LLE) (correlation coefficient 0.095). CONCLUSION: These data suggest that CT underestimates the tortuosity of iliac vessels. Aortic angiography is necessary for accurate AAA endovascular stent design. PMID- 12543565 TI - Endovascular abdominal aortic aneurysm repair outside a tertiary referral centre: feasibility and impact upon workload. AB - In the UK, the majority of endovascular abdominal aortic aneurysm repairs (EVAR) are carried out in tertiary referral centres. We studied the feasibility and impact upon workload of an endovascular programme introduced into a district general hospital. Data was collected prospectively on all patients considered for EVAR since the inception of the programme in April 1999. Evaluation was by contrast enhanced CT scan followed by angiography if potentially suitable for EVAR. We recorded eligibility for EVAR, reasons for exclusion and the outcome of all patients irrespective of the mode of repair. Ninety patients were assessed over a 26-month period, nine of which did not complete the evaluation process. Sixty patients had one or more morphological features that precluded EVAR. Twenty one patients were eligible for EVAR (26%), of which eighteen proceeded to EVAR, one awaits EVAR and two were below the treatment threshold (<5.5 cm). Seventeen had successful aneurysm exclusion and one required immediate conversion. One patient died within 30 days from gastrointestinal haemorrhage. There was one type II endoleak at 1 month and to date we have encountered no ruptures or aneurysm related deaths. Of the remaining 60 patients, 29 have proceeded to elective and 8 to urgent open repair with 2 deaths within 30 days in each group. EVAR is feasible outside tertiary referral centres and satisfactory early results can be achieved. An endovascular programme has important implications upon radiological and surgical workload. The operative mortality of patients undergoing open aortic repair remains acceptable following the introduction of an endovascular service into a district general hospital. PMID- 12543566 TI - Dyeless vascular surgery. AB - PURPOSE: The morbidity associated with contrast-based diagnostics performed for preoperative evaluation prior to vascular intervention ranges from 1 to 21%. These complications range from minor hematomas to death. However, these exams are commonly felt to be a necessary step to completely evaluate the arterial tree before intervention is undertaken. Since this has varied from our experience, we reviewed our experience with repair of abdominal aortic aneurysms (AAAs), carotid endartectomy (CEA), and lower extremity revascularization performed without preoperative contrast studies. MATERIALS AND METHODS: During the last 10 years, we have performed 184 elective AAA repairs with abdominal-pelvis CAT scan without intravenous contrast as a preoperative study. During this same period of time, 903 CEAs were performed in 810 patients based solely on duplex ultrasonography or in combination with magnetic resonance angiography in cases where duplex ultrasonography was inconclusive (53 cases). Finally, over the last 30 months, we have performed 485 revascularizations in the lower extremity based solely on duplex ultrasonography mapping. Direct visualization of all major arteries from the distal aorta to the pedal vessels was performed using duplex imaging. Both the carotid duplex imaging and lower extremity duplex imaging were confirmed to have greater than 95 % positive predictive value during an initial phase of 50 cases confirmed with MRA and contrast angiography respectively. RESULTS: All cases of venous anomalies such as retrocaval left renal vein or left sided inferior vena cava in AAA patients were accurately identified and confirmed by intraoperative findings. No cases of horseshoe kidney were identified. Despite the presence of diminished femoral pulses in six patients, aortic reconstructions were performed with only duplex imaging. The 30 day mortality of AAA patients was 5% for elective repairs. In addition, no gross differences were appreciated with intraoperative findings of CEA as compared to preoperative duplex findings. However, in 5 cases CEA could not be performed due to extension of the lesion well above the available surgical exposure. The 30 day mortality of the CEA patients was 0.7% and the incidence of postoperative stroke or transient ischemic attack was 0.7%. Finally, in two early cases of lower extremity revascularization, the distal anastomosis was placed proximal to a lesion. This was appreciated during the procedure and corrected with a jump graft in each case. CONCLUSIONS: These data suggest that AAA repair, CEA, and lower extremity revascularization can be performed without contrast based preoperative studies and without compromise to evaluation of disease, patient safety or patency of bypass grafts. PMID- 12543567 TI - Visceral artery aneurysms: a single center experience. AB - BACKGROUND: Splanchnic artery areurysm is a rare but clinically relevant disease, showing a high mortality rate in emergency surgery. Reports on splanchnic aneurysms are rare and often anecdotal. The aim of this study is to discuss data obtained from 16 patients in a single vascular surgery center. MATERIALS AND METHODS: Between January 1987 and December 2000, 16 patients underwent surgery for splanchnic artery aneurysms. In 13 patients the localization was single (in two associated with an infrarenal abdominal aortic aneurysm) and in 3 patients multiple. The arteries involved were: splenic (8), hepatic (4), celiac (3), superior mesenteric (3), ileocolic (2), and pancreatoduodenal (1). 13 patients were asymptomatic and 3 presentec with abdominal pain. All patients underwent CT scan, and abdominal aortic and selective visceral artery angiography. Before surgery, all patients underwent cerebral MR or cerebral CT scan. 13 patients underwent open vascular surgery; 3 patients (2 splenic and 1 hepatic) underwent endovascular procedure (angioembolization). Histological examination of the aneurysmal wall was obtained in 14 patients. Ultrasound examination was performed after 6 months, then yearly. 14 patients underwent abdominal CT scan during the long-term follow-up. RESULTS: Perioperative mortality was absent. 12 cases were classified as displastic, with minor or major secondary atherosclerotic changes, and in many cases severe calcications. 2 cases were classified as atherosclerotic. Cerebral MR did not show any cases of intracerebral displastric aneurismal disease. One patient was lost at follow-up after 9 years. One patient showed a recurrence at 6 years (superior mesenteric artery): the patient underwent a new surgical procedure and died 20 days after surgery for intestinal infarction. All abdominal follow-up CT scans show good results of the vascular reconstruction and escluded other new visceral or aortic aneurysms. CONCLUSIONS: Visceral artery aneurysms are an uncommon form of abdominal vascular disease showing a high postoperative mortality rate in emergencies. Surgery, and in selected cases, endovascular treatment, can successfully manage splanchnic artery aneurysms with few complications and low recurrence. PMID- 12543568 TI - Intra-operative assessment of technical defects after carotid endarterectomy: a comparison between angiography and colour duplex scan. AB - We compared the intra-operative diagnostic value of CDS vs IA-DSA in identifying major and minor technical defects after CEA. Between August 1997 and December 1998, 138 consecutive patients undergoing 141 carotid endarterectomies were intra operatively investigated with colour duplex scan and intra-arterial digital subtraction angiography. Thirty-six (25.5%) technical defects were identified. Four (11.1%) major defects were detected by both methods and they were immediately corrected. Fifteen (41.6%) minor defects were detected by both methods, thirteen (36.1%) minor defects were detected by colour duplex but ignored by angiography. Angiography detected four (11.1%) kinkings missed with the colour duplex. The overall sensitivity of both methods for major defect was 100%. The sensitivity of colour duplex for minor defects was 87% vs 59% for angiography. On the basis of our study, colour duplex could be considered the choice method for quality control after carotid endarterectomy. PMID- 12543569 TI - The chemically treated bovine ureter--clinical performance of a novel biological vascular prosthesis. AB - AIM: This paper describes the rationale, development and clinical performance of a novel small-diameter vascular prosthesis of non-vascular animal tissue origin. METHOD: Durable conduits suitable for coronary and peripheral arterial reconstruction are prepared for the first time from non-vascular animal tissue. Australian bovine ureters from young steers from protected herds are processed to form strong, bloodflow-compatible and non-immunogenic biologic grafts ('Flonova', Bionova International Pty Ltd, Melbourne). Fifty-six patients lacking suitable autogenous veins received 62 prostheses (32 bypasses were implanted above knee and 30 below knee). Determination of patency and function included Doppler ultrasound, duplex scanning and angiography. FINDINGS: The primary patency rate at 1, 2 and 5 years, respectively was 67, 59 and 52% and the secondary patency for the same time frames was 88, 73 and 61%. There were no occlusions as a result of intimal hyperplasia. Dilatation of the wall was diagnosed in one prosthesis at 3.5 years, and one at 7 years, and a false aneurysm of unknown aetiology occurred in another at 3 months. There were no primary graft infections. CONCLUSION: This evaluation shows the chemically treated bovine ureter (CTBU) is a viable medium term alternative to the currently available artificial vascular prostheses for peripheral revascularisation. PMID- 12543570 TI - Primary forearm arteriovenous fistula for hemodialysis access--an integrated approach to improve outcomes. AB - PURPOSE: Maximizing the ratio of primary arteriovenous fistula (PAVF) over bridge graft fistula (BGF) for hemodialysis access is a primary recommendation of the National Kidney Foundation published as Dialysis Outcomes Quality Initiative (DOQI). Imaging, anesthetic and surgical techniques were taken into account to achieve this and other goals, including extensive use of forearm vessels to lower immediate and early failure rates and prolong the useful life of PAVFs. DESIGN: Prospective non-randomized study. METHODS: High-resolution duplex ultrasonography (DUS) was added to careful clinical assessment in planning and follow-up of the dialysis access. Brachial plexus block, which allowed the use of an arterial tourniquet and gave a postoperative sympathectomy type effect, was used for anesthesia, and together with meticulous surgical technique, prevented spasm. Access puncture, post-operative follow-up and surgical revisions were planned in close cooperation with the nephrology team. FINDINGS: Ninety (57.3%) of the 157 fistulas constructed for new hemodialysis access between August 1998 and March 2000 were PAVFs. Seventy-three (81.1%) of these were confined to the forearm and comprise the study population, with a mean follow-up of 8.4+/-4.4 months. There were no immediate failures in the study group. The early failure rate (1 month) was 6.8% and revisions based on DUS were easily accomplished in all cases. The one year assisted primary patency rate was 81.8% and the secondary patency rate at 18 months was 98.6%. CONCLUSIONS: DUS for planning and follow-up of PAVF along with careful surgical technique under a brachial plexus blockade can achieve a PAVF/BGF ratio well above 50% with a low early failure rate and a high secondary patency rate. Algorithms are presented to achieve these goals. PMID- 12543571 TI - Pelviureteric junction disruption as a complication of chemical lumbar sympathectomy. AB - Chemical lumbar sympathectomy is a commonly performed procedure in vascular surgery and pain management. This case report discusses the management of a patient who suffered pelviureteric junction disruption following phenol injection for ischaemic leg pain despite radiological evidence of correct placement. The authors suspect this is an underreported complication, which could be relevant in obtaining informed consent. PMID- 12543572 TI - The effect of inhibition of renin-angiotensin system by valsartan during hypovolemic shock and low flow sigmoideal ischaemia in pigs. AB - The aim of this experiment was to study the effect of Renin-Angiotensin System (RAS) blockade by means of valsartan on the colonic and systemic circulation in pigs during low flow sigmoideal ischemia in combination with hypovolemic shock. This condition resembles the situation that occurs in patients suffering from a ruptured aneurysm and a compromised colonic circulation. An experimental study in pigs was performed : 6 pigs with low flow sigmoideal ischemia and hypovolemic shock were treated with valsartan and a control group of 5 pigs with low flow sigmoideal ischemia and hypovolemic shock without medical treatment.Valsartan, 3 mg/kg, was administered intravenously. The operation was performed via left sided lumbotomy. The distal aorta was partially occluded to a flow reduction of 30% of the initial value. Hypovolemic shock was induced by withdrawing 20 ml/kg blood in 45 min. Resuscitation with 30 ml/kg haemaccel was iniated after 2 h of shock. The following parameters were measured: blood pressure, cardiac output; hemoglobin, lactate, angiotensin II in mixed venous blood (obtained from pulmonary artery) and in splanchnic blood (obtained from caudal mesenteric vein); and endoluminal pulse oximetry of the sigmoideal mucosa. Statistical analysis was performed by ANOVA and Wilcoxon signed rank test. There was a significant increase of lactate levels both in systemic and splanchnic circulation (P<0.05) in both groups. In the control group, the mean angiotensin II concentrations in the systemic circulation increased, after induction of ischaemia and shock. In the experimental group, the increase in angiotensin concentrations after resuscitation was significantly more prominent. In the colonic circulation, in both groups, there was a significant increase in angiotensin II levels in the splanchnic circulation following ischaemia and reperfusion (P<0.05), but there was no significant difference between the groups. There were no detectable mucosal signals measured by pulse oximetry after induction of shock throughout the experiment, whereas in the experimental group, median mucosal oxygen saturations of 81, 74.5 and 85% were achieved after resuscitation and declamping (P<0.01).In conclusion, angiotensin II inhibition during hypovolemic shock improves the colonic circulation, measured by pulse oximetry. However, other parameters of tissue ischaemia did not improve. PMID- 12543573 TI - Fluvastatin induces apoptosis of vascular endothelial cells: blockade by glucocorticoids. AB - Statins block de novo synthesis of cholesterol by inhibiting the enzyme, HMG CoA reductase. The product of this reaction, mevalonic acid, is also a precursor of isoprenoids, molecules required for the activation of signaling G-proteins, such as Ras. Signal transduction pathways involving Ras are important for cell survival and this may be why statins induce apoptotic death of several cell types. Given that statins are used to treat vascular disease, surprisingly no studies have been conducted on vascular endothelial cells. Here we show that fluvastatin (FS), at concentrations from 1-2 microM, blocks growth and induces apoptosis of the endothelial cell line, EA.hy 926. Considerable redundancy is known to exist in cell signaling and in vivo toxicity of FS might be prevented by other signaling pathways, like those activated by adrenal or sex steroids. RT-PCR analysis revealed the expression of the androgen and glucocorticoid receptor in EA.hy 926 cells. Although the androgen, dihydrotestesterone (DHT) had no effect, the glucocorticoid, dexamethasone (Dex), blocked FS-induced apoptosis. Cell cycle analysis revealed that 24 h exposure to FS prevented cells from leaving G(1) and 24-48 h later a marked sub-G(1) peak was observed. Dex was able to reduce the sub G(1) peak, but it failed to block accumulation of cells in G(1), indicating that it's effect was specific for blockade of apoptosis, and not specific to an effect on FS alone. This study strongly suggests that glucocorticoids have a role to play in preventing vascular injury and they may provide the reason why statins are not inherently toxic to vascular endothelial cells, in vivo. PMID- 12543574 TI - Undiagnosed aberrant right subclavian artery: pitfall in aortic arch surgery. AB - Selected cerebral perfusion as brain protection via right subclavian artery during aortic arch aneurysm repair adds safety, but may be jeopardized by aortic arch anomalies not readily recognized preoperatively. We describe a case of transverse aortic arch aneurysm repair where an undiagnosed aberrant right subclavian artery was cannulated for selective brain protection. PMID- 12543575 TI - Issues concerning consent in patients undergoing cardiac surgery-the need for patient-directed improvements: a UK perspective. AB - AIMS: To determine the extent to which informed patient consent for cardiac surgery corresponds with standard guidelines and to assess patient satisfaction with the consent process. METHODS: A questionnaire developed in accordance with consent guidelines published by the Medical Defence Union (MDU) and the Practicalities Of Producing Patient information (POPPi) Guide was administered to 82 patients after they were consented for cardiac surgery. RESULTS: Patients were informed about i) type of operation [n=80, 97.5%] ii) reason for surgery [n=78, 95%] iii) surgical procedure [n=77, 94%] iv) length of hospital stay [n=73, 89%] and v) post-surgical care [n=79, 96.3%]. However, a decreasing number of patients were notified about i) possible risks of surgery [n=64, 78%] and ii) consequences of no surgery [n=62, 75.6%]. Interestingly, patients received less adequate information about i) alternative methods of treatment (n=34, 41.4%], ii) future lifestyle (n=55, 67%] and iii) medications following surgery [n=40, 48.7%]. 81% (n=67) were overall 'satisfied' with the information provided. More than 95% (n=78) patients were invited to ask questions. The most commonly asked questions were about i) quality of life ii) length of operation and hospital stay iii) surgical procedure iv) consequences of no surgery. CONCLUSIONS: The study has highlighted areas of informed consent in cardiac surgical patients that need attention. Although, patients were generally satisfied and well informed about the procedure, less information was presented covering alternative treatments, future lifestyle and medications. This was reflected in the questions asked by patients and their suggested improvements for the consent procedure. PMID- 12543576 TI - The Fontan-operation: from intra- to extracardiac procedure. AB - PURPOSE: For treatment of univentricular heart, the Fontan operation has been established as the definitive palliation. The current controversy is mainly based on the high incidence of arrhythmias after an intra-atrial lateral tunnel Fontan operation. METHODS: From January 1995 until April 2002, 46 children underwent a Fontan-type operation with or without a small fenestration. In 33 patients (group I) an intracardiac tunnel and in 13 patients (group II) an extracardiac conduit procedure was performed. PRINCIPAL FINDINGS: There was no perioperative mortality. All patients showed postoperative a significant increase of arterial oxygen saturation, from 76 to 86% after surgery with fenestration, or to 90.5% without fenestration respectively. In patients with fenestration procedure, the saturation rose to 90% after closure of fenestrations 9 to 12 months after operation. CONCLUSIONS: Modified Fontan operations can be performed in normothermia on the beating heart with acceptable mortality. The extracardiac conduit Fontan procedure has the benefits of less surgical injury and a higher intraoperative flexibility. PMID- 12543577 TI - Off-pump coronary surgery: surgical strategy for the high-risk patient. AB - OBJECTIVE: In a retrospective study, we compared two groups of consecutive patients operated by the same team during the year 2000 for coronary artery disease with the use of extracorporeal circulation (group 1, n=230) or on the beating heart using the Octopus II plus stabiliser (group 2, n=228). High-risk patients were identified by a EuroSCORE plus 6. EuroSCORE definitions and predicted risk models were utilized to compare the variables of the groups. METHODS: There were no significant differences between the preoperative variables of the groups in age, gender, left ventricular function, diabetes and peripheral vascular and renal disease as is indicated by the Euroscore (resp. 4.7/5.1 p=0.107). Calcification of the ascending aorta and chronic obstructive lung disease were statistically significant more prevalent in the beating heart group. No differences in preoperative variables in the high-risk patients group (Euroscore 8.5/8.1 p=0.356) except for calcification of the ascending aorta. RESULTS: All patients underwent a full revascularisation through a midline sternotomy. Significant more distal anastomoses were performed in group 1 (3.7 per patient (1-6)) with regard to group 2 (2.9 per patient (1-6)). Anesthesia, postoperative treatment and follow up were equal for both groups. A significant lower incidence of atrial fibrillation (p=0.010), shorter ICU stay (p=0.031) and renal insufficiency (p=0.033) was reported in group 2. In the low risk group, we could not diagnose any difference between the two groups, except for atrial fibrillation. The benefits of the beating heart surgery however were more pronounced in the high-risk patient as is indicated by a significant reduction of the ICU stay by 1 day (3.5d/2.5d (p=0.028)), better preservation of the renal function (p=0.017) and a significant reduction of the length of hospital stay by more than two days (p=0.040). A lower incidence of atrial fibrillation, however not significant. CONCLUSION: In our experience, beating heart surgery is a safe alternative for conventional coronary heart surgery. High-risk patients do benefit most from this technique. It became our first choice in the elderly patient and patients presenting with higher co-morbidities. PMID- 12543578 TI - An experimental model of coronary anastomosis without suturing. AB - OBJECTIVE: The aim of the study is to explore the feasibility and mid-term patency of an easier anastomotic technique for Minimally Invasive Direct Coronary Bypass Grafting (MIDCAB). METHODS: Eight mongrel dogs (+/-15 kg) underwent direct anastomosis between the left internal thoracic artery (LITA) and the left anterior descending coronary artery (LAD) via inferior sternotomy on the beating heart. After positioning the graft, the distal part of the LAD was opened to allow retrograde filling of the LITA-graft. The anastomosis was secured by the use of biological glue (BioGlue, Cryolife, Marietta, GA, USA). No intravascular suture material was used. Ischemic time averaged 6 min. The proximal LAD was occluded upstream the arteriotomy. All survivors were angiographically controlled for patency after 6-8 weeks. Consequently, four dogs were sacrificed after 6 weeks and the remaining after 3 months for anatomo-pathological and histological examination by light and electron microscopy of the anastomotic site. RESULTS: All procedures were successful except for one animal that died of uncontrollable bleeding at the anastomotic site. Another sustained post-operative transmural anterior myocardial infarction due to a late graft occlusion. All angiographically controlled grafts were patent with two vascular strings near the anastomotic site. Histology showed early macrophage infiltration into the glue. At post-mortem examination, new endothelialization was noticed in 80% of the cases. However, ultrastructural examination detected marked differences in endothelial fibroblastic lining compared to normal histology. CONCLUSIONS: Good mid-term permeability of the LITA grafts was observed in this new anastomotic technique for MIDCAB in the canine model. Although neo-endothelialization was present in most cases, ultrastructural differences were noticed after 3 months in the neo-intima compared to normal. PMID- 12543579 TI - Preliminary evaluation of the Sternum Screw: a novel method for improved sternal closure to prevent dehiscence. AB - Sternal dehiscence is a relatively rare but serious complication of sternal closure with an unacceptable mortality and morbidity rate. The 6 mm cannulated Sternum Screws are a novel approach that aims to prevent dehiscence. The screws are placed on either side of the sternotomy and wire threaded through the cannula and closed customarily. The Sternum Screws make the bone non-penetrable at the same time retaining the familiarity of conventional stainless steel wire. This novel technique was tested in sheep sterna to compare its efficacy to conventional wire closure. Using tensile testing in a testing machine, randomised controlled closures of the Sternum Screws and No. 5 stainless steel wires were evaluated until system failure. Seventeen matched pairs were tested. The Sternum Screw closure was on average 36% stronger, 284+/-43 N (mean+/-SD) compared to conventional wire closure alone, 215+/-38 N (mean+/-SD) [p<0.0001 by t-test]. System failure in 82% of Sternum Screw closures, however, was due to wire breakage or untwisting rather than the screw itself. In all these cases the screw remained intact in bone. 71% of conventional wire closures failed by dehiscing through the sterna. The mean forces required for wire dehiscence and wire failure in the Sternum Screws may be achieved in vivo during large coughs. The study shows there is merit in further evaluating the approach as a method of preventing dehiscence. It also highlights the use of alternative wiring techniques increase the tensile strength of the closure yet retain the familiarity and versatility of conventional wire. PMID- 12543580 TI - Two cases of anomalous origin of LAD from right coronary artery requiring coronary artery bypass. AB - We experienced two cases with anomalous origin of the left anterior descending artery (LAD) from the proximal right coronary artery requiring coronary artery bypass grafting. A 66-yr old female with a long history of angina and a positive stress test had the anomalous artery coursing anterior to the right ventricular outflow tract. A 42-yr old male with worsening angina after an anteroseptal myocardial infarction had the anomalous artery running between the great vessels. Both patients underwent left internal mammary artery-to-LAD bypass on the beating heart with complete resolution of ischemic symptoms. Isolated coronary artery anomaly is an uncommon disease (0.6-1.2%) in patients undergoing cardiac catheterization. An anomalous origin of the LAD from the proximal right coronary artery (RCA) or the right sinus of Valsalva (RSV) is very rare, found in 1.2-6.1% of all coronary anomalies. This coronary anomaly has been considered potentially serious but functionally unimportant. We report two cases of anomalous LAD from the proximal RCA resulting in anterior wall ischemia which was effectively treated by coronary artery bypass surgery. PMID- 12543581 TI - Treatment of an aortic fungal false aneurysm by composite stentless porcine/pericardial conduit: a case report. AB - Fungal prosthetic valve endocarditis is an uncommon but serious condition with high early and long-term mortality. The majority of these cases occur after aortic valve surgery and are caused by Candida species. Radical debridement of all infected tissues, valve replacement with perioperative and long-term anti fungal agents is the recommended treatment. Choice of prosthesis varies widely among surgeons, but present recommendations favour biological prostheses. We report for the first time the case of a fungal PVE with false aneurysm after composite aortic root replacement with a dacron composite conduit treated successfully with aortic root replacement using a Shelhigh (Shelhigh Inc., Millburn, NJ) stentless porcine pericardial valved conduit. PMID- 12543582 TI - A thymic carcinoid mimicking acute aortic dissection. AB - Thymic carcinoids are extremely rare. We present an 80 year old hypertensive patient who presented as an emergency with sudden onset of severe chest and upper back pain raising the suspicion of acute aortic dissection or acute coronary syndrome. Further investigations revealed a carcinoid tumour of thymic origin. PMID- 12543587 TI - Three for 2003: reducing the burden of stress. PMID- 12543588 TI - Postmenopausal hormones: where do we go from here? PMID- 12543589 TI - New ways to control psoriasis. PMID- 12543590 TI - Antiperspirants don't cause breast cancer. PMID- 12543591 TI - Waist better than weight as measure of health risk. PMID- 12543592 TI - By the way doctor. I've often wondered about the expiration data on prescription drugs. Is it unsafe to take them after they expire? PMID- 12543593 TI - By the way, doctor. What can you tell me about the new drug Provigil? I've read that it helps people stay awake without feeling jittery. For those of us who work a lot of overtime, it sounds like a dream come true. PMID- 12543594 TI - Diet wars II: how do the plans measure up? PMID- 12543595 TI - Viagra and erections: do the benefits persist? PMID- 12543596 TI - Snow shoveling: The cold facts. PMID- 12543597 TI - On call. I have arthritis of both knees. Prescription medicines didn't help and they irritated my stomach. I tried glucosamine, and it didn't do much good either. Finally my doctor sent me to a specialist, who washed out both knees with an arthroscope. My pain is nearly gone, but I read that doctors did a test that showed a fake operation was just as good as my surgery. It's an outrage. How can responsible doctors perform fake operations? PMID- 12543598 TI - On call. My wife and I are both 62 and healthy. She started taking Prempro during her menopause eight years ago but has now decided to stop the hormones. I began taking DHEA five years ago, but I switched to AndroGel when my doctor gave me a prescription last year. Should I stay on AndroGel, go back to DHEA, or stop hormones? PMID- 12543600 TI - How autism looks. PMID- 12543599 TI - Generalized anxiety disorder: toxic worry. PMID- 12543601 TI - Scared crooked. PMID- 12543602 TI - Mental illness and medical care. PMID- 12543603 TI - The truth about mold. Most experts say there's more fear than fact to "toxic mold." But that doesn't mean that indoor mold can't cause health problems. PMID- 12543604 TI - Hooked on fish? There might be some catches. Health-conscious people eat it three, even four times a week. But farm-raised fish and worries about mercury contamination are churning the waters. PMID- 12543605 TI - Men's health: a conversation with an expert. PMID- 12543606 TI - Grapefruit juice and your medications. PMID- 12543608 TI - Killer snow. PMID- 12543607 TI - Picky people, nosey germs. PMID- 12543609 TI - By the way, doctor. I have diabetes and two years ago I started to have pain in my feet with numbness and tingling. My doctors say that the problem is damage to my nerves related to the diabetes. Any ideas? PMID- 12543610 TI - By the way doctor. Is there a best time to take prescribed cardiac pills? Does a multivitamin or a calcium pill cancel out their effects? PMID- 12543611 TI - Keeping plaque in its place. New therapies focus on stabilizing artery-clogging plaque rather than getting rid of it. PMID- 12543612 TI - 7 ways to save money on prescription drugs. PMID- 12543613 TI - A shock at home can save a life. For some people, it makes sense to have an automated external defibrillator at home. PMID- 12543614 TI - Ask the doctor. Every so often I get a feeling of pressure around my heart that can last for hours. It worries me since I've already had several procedures for coronary artery disease. Can episodes of angina last this long? PMID- 12543615 TI - Ask the doctor. A heart attack caused me to develop heart rhythm problems. These made it necessary for me to get an implantable cardioventer/defibrillator (ICD) that can shock my heart back to a normal rhythm. My medical story is a lot like Vice President Dick Cheney's, but my social situation isn't--I don't have anyone to drive me around. My doctor doesn't want me to drive. But depending on friends and family is a major problem, and we don't live near public transportation. Any suggestions? PMID- 12543616 TI - Snippets from the past: 70 years ago in the journal. PMID- 12543617 TI - Risk factors for ectopic pregnancy: a comprehensive analysis based on a large case-control, population-based study in France. AB - This case-control study was associated with a regional register of ectopic pregnancy between 1993 and 2000 in France. It included 803 cases of ectopic pregnancy and 1,683 deliveries and was powerful enough to investigate all ectopic pregnancy risk factors. The main risk factors were infectious history (adjusted attributable risk = 0.33; adjusted odds ratio for previous pelvic infectious disease = 3.4, 95% percent confidence interval (CI): 2.4, 5.0) and smoking (adjusted attributable risk = 0.35; adjusted odds ratio = 3.9, 95% CI: 2.6, 5.9 for >20 cigarettes/day vs. women who had never smoked). The other risk factors were age (associated per se with a risk of ectopic pregnancy), prior spontaneous abortions, history of infertility, and previous use of an intrauterine device. Prior medical induced abortion was associated with a risk of ectopic pregnancy (adjusted odds ratio = 2.8, 95% CI: 1.1, 7.2); no such association was observed for surgical abortion (adjusted odds ratio = 1.1, 95% CI: 0.8, 1.6). The total attributable risk of all the factors investigated was 0.76. As close associations were found between ectopic pregnancy and infertility and between ectopic pregnancy and spontaneous abortion, further research into ectopic pregnancy should focus on risk factors common to these conditions. In terms of public health, increasing awareness of the effects of smoking may be useful for ectopic pregnancy prevention. PMID- 12543618 TI - Subfecundity as a correlate of preeclampsia: a study within the Danish National Birth Cohort. AB - A long interpregnancy interval is associated with preeclampsia. If some women experiencing a long interval between births had difficulty conceiving, subfecundity and preeclampsia may share a common etiology. Therefore, the authors examined the association between subfecundity and preeclampsia. By using interview data collected during the second trimester of pregnancy (1998-2001) from women participating in the Danish National Birth Cohort, they identified 20,034 and 24,698 singleton livebirths to primiparous and multiparous women, respectively, for whom preeclampsia information was available from hospital birth records. Among women with no known hypertension, the authors estimated a higher risk of preeclampsia in those with longer times to pregnancy (TTPs), after adjustment for maternal age, prepregnancy body mass index, and smoking. Compared with primiparas who became pregnant right away (referent category), the risk of preeclampsia increased with TTP and then stabilized for women taking 6 months or longer to conceive, whose risk of preeclampsia increased by 50%. Multiparas also had an increased risk, but only those reporting a TTP longer than 12 months (odds ratio = 2.47, 95% confidence interval: 1.30, 4.69). The authors found that a long TTP was associated with preeclampsia, supporting the hypothesis that some factors delaying clinically recognized conception may also be in a causal pathway for preeclampsia. PMID- 12543619 TI - Maternally transmitted antibodies to pregnancy-associated variant antigens on the surface of erythrocytes infected with Plasmodium falciparum: relation to child susceptibility to malaria. AB - The consequences of pregnancy-associated malaria on a child's health have been poorly investigated. Malarial infection of the placenta seems to result in a higher susceptibility of children to the parasite during their first year of life. In 1993-1995, the authors investigated the role of antibodies in variable surface antigens (VSA) specific to chondroitin sulfate A (CSA)-binding parasites to assess the parasitologic status of the child. Flow cytometry was used to measure levels of antibodies to VSA from CSA-selected and -unselected parasite lines in the cord blood of 79 newborns in Ebolowa, Cameroon. These newborns were subsequently followed up for 2 years to determine the date of first occurrence of blood parasites and mean parasite density during follow-up. Maternally transmitted antibodies to VSA expressed by CSA-binding parasites, but not antibodies to any other specificity, were negatively related to time of first appearance of Plasmodium falciparum in a child's blood and were positively related to mean parasite density during the first 2 years of life. If maternal infection is thought to be the main mechanism influencing susceptibility of the newborn to malaria, antibodies to VSA may better denote maternal malaria infection during almost the entire pregnancy as opposed to placental blood smear reflecting infection during the last few months of pregnancy. PMID- 12543620 TI - Delayed application of condoms is a risk factor for human immunodeficiency virus infection among homosexual and bisexual men. AB - The objective of this analysis was to identify risk factors for recent human immunodeficiency virus (HIV) infection among homosexual and bisexual men in Ontario, Canada, during 1998-2001. Participants were recruited through the provincial HIV diagnostic laboratory and through physicians and community organizations. HIV test results were used to identify recent seroconverters (cases). A subsample of 183 men (62 cases and 121 controls) enrolled in the Polaris HIV Seroconversion Study as of June 2001 was analyzed. This analysis focused on sexual behaviors with partners who were HIV-positive or whose HIV status was unknown. In multiple logistic regression, independent risk factors were identified. Rates of unprotected receptive oral, insertive anal, and receptive anal sex and delayed application of condoms during receptive anal sex (RAS) were significantly higher among cases (97%, 41%, 53%, and 32%, respectively) than among controls (73%, 19%, 14%, and 2%). Independent risk factors for HIV infection were RAS without condoms (odds ratio = 4.4, p = 0.0004) and delayed application of condoms during RAS (odds ratio = 5.8, p = 0.01). There was an association with condom failure (breakage or slippage) during RAS that approached significance (odds ratio = 2.9, p = 0.09). Delayed application of condoms for RAS may result in contact with preejaculatory fluid. This behavior, which to date has received little attention, may pose as much risk for HIV infection as fully unprotected RAS. PMID- 12543621 TI - Genital human papillomavirus infection: incidence and risk factors in a cohort of female university students. AB - Incidence data on human papillomavirus (HPV) infection are limited, and risk factors for transmission are largely unknown. The authors followed 603 female university students in Washington State at 4-month intervals between 1990 and 2000. At each visit, a sexual and health questionnaire was completed and cervical and vulvovaginal samples were collected to detect HPV DNA. At 24 months, the cumulative incidence of first-time infection was 32.3% (95% confidence interval: 28.0, 37.1). Incidences calculated from time of new-partner acquisition were comparable for enrolled virgins and nonvirgins. Smoking, oral contraceptive use, and report of a new male sex partner--in particular, one known for less than 8 months before sex occurred or one reporting other partners--were predictive of incident infection. Always using male condoms with a new partner was not protective. Infection in virgins was rare, but any type of nonpenetrative sexual contact was associated with an increased risk. Detection of oral HPV was rare and was not associated with oral-penile contact. The data show that the incidence of HPV associated with acquisition of a new sex partner is high and that nonpenetrative sexual contact is a plausible route of transmission in virgins. PMID- 12543622 TI - Exposure to metal fume and infectious pneumonia. AB - To test the hypothesis that inhalation of metal fume reversibly increases susceptibility to pneumonia, the authors conducted a case-control study. Men aged 20-64 years, admitted to 11 hospitals in West Midlands, England, with community acquired pneumonia during 1996-1999 were interviewed about their lifetime occupational history, exposure to metal fume, and potential confounding factors. Similar information was collected from controls admitted to the same hospitals with nonrespiratory illness. For cases, exposures were timed relative to the onset of their illness (on average, 6 months before interview). Exposure histories for controls were censored 6 months before interview. Interviews were completed by 525 cases and 1,122 controls (response rates of 74% and 99%). Pneumonia was associated with reported occupational exposure to metal fume in the previous year (adjusted odds ratio (OR) = 1.6, 95% confidence interval (CI): 1.1, 2.4) but not in earlier periods (OR = 1.1). The risk was highest for lobar pneumonia and recent exposure to ferrous fume (OR = 2.3, 95% CI: 1.2, 4.3). The association was not specific to any one microorganism. These findings support the hypothesis that ferrous and possibly other metal fumes reversibly predispose to infectious pneumonia. Research should now focus on the underlying mechanisms and prevention. PMID- 12543623 TI - Shaving, coronary heart disease, and stroke: the Caerphilly Study. AB - The relation between frequency of shaving and all-cause and cardiovascular disease mortality, coronary heart disease, and stroke events was investigated in a cohort of 2,438 men aged 45-59 years. The one fifth (n = 521, 21.4%) of men who shaved less frequently than daily were shorter, were less likely to be married, had a lower frequency of orgasm, and were more likely to smoke, to have angina, and to work in manual occupations than other men. Over the 20-year follow-up period from 1979-1983 to December 31, 2000, 835 men (34.3%) died. Of those who shaved less frequently than daily, 45.1% died, as compared with 31.3% among those who shaved at least daily. Men who shaved less frequently had fully adjusted hazard ratios (adjusted for testosterone, markers of insulin resistance, social factors, lifestyle, and baseline coronary heart disease) of 1.24 (95% confidence interval (CI): 1.03, 1.50) for all-cause mortality, 1.30 (95% CI: 0.99, 1.71) for cardiovascular disease mortality, 1.08 (95% CI: 0.61, 1.92) for lung cancer mortality, 1.16 (95% CI: 0.90, 1.48) for coronary heart disease events, and 1.68 (95% CI: 1.16, 2.44) for stroke events. The association between infrequent shaving and all-cause and cardiovascular disease mortality is probably due to confounding by smoking and social factors, but a small hormonal effect may exist. The relation with stroke events remains unexplained by smoking or social factors. PMID- 12543624 TI - Mortality from diseases of the circulatory system in radiologic technologists in the United States. AB - Although increased mortality from diseases of the circulatory system has been observed in patients treated with radiotherapy, the effects of chronic low-dose radiation exposure are not clear. Among 90,284 US radiologic technologists who responded to a mailed questionnaire during 1983-1989, the authors evaluated mortality from circulatory system diseases through 1997 in relation to job history and work procedures as surrogates for radiation exposure. They used Poisson regression models stratified for sex, race, age, and calendar year and adjusted for smoking, body mass index, alcohol intake, marital status, parity, menopausal status, and history of myocardial infarction. A total of 1,107,100 person-years accrued, and 1,070 subjects died from circulatory system diseases. Relative risks for first employment during 1950-1959, 1940-1949, or before 1940, compared with 1960 and later, were 1.01 (95% confidence interval (CI): 0.78, 1.30), 1.14 (95% CI: 0.86, 1.50), and 1.42 (95% CI: 1.04, 1.94), respectively (trend p < 0.001). For the subset of deaths from cerebrovascular disease (n = 174), the respective relative risks were 0.90 (95% CI: 0.45, 1.78), 1.54 (95% CI: 0.74, 3.23), and 2.40 (95% CI: 1.09, 5.31) (trend p = 0.004), and for deaths from ischemic heart disease (n = 633), the relative risks were 0.98 (95% CI: 0.71, 1.35), 1.00 (95% CI: 0.71, 1.42), and 1.22 (95% CI: 0.81, 1.82) (trend p = 0.026). The relative risks for mortality from circulatory system diseases and the subset of cerebrovascular disease increased significantly with the number of years worked before 1950 (trend p = 0.007 and < 0.001, respectively). The data suggest increased mortality from diseases of the circulatory system with occupational radiation exposure before 1950 when radiation doses were likely high. PMID- 12543625 TI - Multiple informants: a new method to assess breast cancer patients' comorbidity. AB - Past assessments of comorbidity indices have sought to recommend a single index that performs better than others. The authors used a multiple informants approach as an alternative method to simultaneously assess five indices of comorbidity. This approach provides a single estimate of the overall effect of comorbidity and evaluates the relation any individual index has to the outcomes of interest. Association of comorbidity with definitive primary therapy, discussion of tamoxifen, and receipt of tamoxifen was evaluated in a cohort of 830 older breast cancer patients enrolled at four geographically distinct centers in the United States from 1996 to 1999. The estimated adjusted effect of a unit increase in comorbidity on the odds of discussing tamoxifen therapy was 0.70 (95% confidence interval: 0.56, 0.88). An increase in comorbidity was not associated with receipt of definitive primary therapy (odds ratio = 0.94, 95% confidence interval: 0.79, 1.13) or receipt of tamoxifen (odds ratio = 0.96, 95% confidence interval: 0.72, 1.27). The multiple informants regression proved superior to separate regression models that included only one index. In analyses that require comorbidity adjustment and for which no single index is expected to be ideal, the multiple informants approach is an attractive alternative to selecting a single index and to other methods of using multiple indices. PMID- 12543626 TI - Accuracy of fetal growth indicators as surrogate measures of steroid hormone levels during pregnancy. AB - This study evaluates the use of fetal growth characteristics as surrogate measures for steroid hormone exposures during pregnancy. The validity of using birth weight, birth length, ponderal index, and placental weight as indicators of pregnancy hormone exposures was assessed using third-trimester serum samples from 568 pregnant women who participated in the Child Health and Development Studies, Berkeley, California (1959-1966). The magnitude of the associations between birth characteristics and hormone concentrations was assessed using geometric means, Pearson's correlations, and linear and logistic regression. Accuracy was evaluated using sensitivity, specificity, and receiver operating characteristic curve analyses. The strongest and most consistent association observed was between birth weight and estriol levels. Despite a positive correlation (r = 0.32) and strong associations with high estriol levels (odds ratio for highest compared with lowest birth weight quartile = 6.63, 95% confidence interval: 3.20, 12.5), the predictive performance of birth weight as a proxy for estriol levels was poor (area under the receiver operating characteristic curve = 0.66, 95% confidence interval: 0.61, 0.71). Likewise, all fetal growth measures revealed little discriminatory ability as indicators of estriol, estrone, estradiol, or progesterone levels. Thus, observed associations with these surrogate measures may not be a reflection of pregnancy hormone exposure and should be interpreted with caution. PMID- 12543627 TI - Validity of self-reported mammography: examining recall and covariates among older women in a Health Maintenance Organization. AB - Self-reports of screening are frequently used in place of chart abstraction, particularly in outpatient settings, because they are generally less expensive and frequently provide the only information available. The authors expanded the literature on validation of self-reported mammography by including the validity of recall and by assessing covariates in a setting where women were examined more than once. In 1995, this study validated mammography use in a sample of 949 women aged 50-80 years who were members of a health maintenance organization with centralized automated records of mammographic examinations. The majority of women had had a mammogram within the previous 2 years according to self-reports and records, but self-reported rates exceeded record rates by 8.2%. Sensitivity was high (93.8%), whereas specificity was low (53.6%). The overall agreement between self-reports and records was 82.7%. The kappa value was 0.52, indicating fair agreement beyond chance. Modeling with logistic regression revealed that being a college graduate and having a first-degree relative with breast cancer were significantly associated with accurate recall. Comparison of actual time interval data revealed that disagreements consisted largely of women's underestimates of time since their last screening. These results add to knowledge about the validity of self-reported mammographic screening data in settings where women are screened more than once. PMID- 12543628 TI - Methodological issues in the surveillance of poisoning, illicit drug overdose, and heroin overdose deaths in new Mexico. AB - New Mexico leads the nation in poisoning mortality, which has increased during the 1990s in New Mexico and the United States. Most of this increase has been due to unintentional deaths from illicit drug overdoses. Medical examiner and/or vital statistics data have been used to track poisoning deaths. In this study, the authors linked medical examiner and vital statistics records on underlying cause of death, coded using the International Classification of Diseases, Ninth Revision, to assess the extent to which these data sources agreed with respect to poisoning deaths. The authors used multiple-cause files, which are files with several causes listed for each death, to further assess poisoning deaths involving more than one drug. Using vital statistics or medical examiner records, 94.7% of poisoning deaths were captured by each source alone. For unintentional illicit drug and heroin overdose deaths, each data source alone captured smaller percentages of deaths. Deaths coded as E858.8 (unintentional poisoning due to other drugs) require linkage with medical examiner or multiple-cause records, because this code identifies a significant percentage of illicit drug overdose deaths but obscures the specific drug(s) involved. Surveillance of poisoning death should include the use of medical examiner records and underlying- and multiple-cause vital statistics records. PMID- 12543630 TI - Re: "Cancer in Korean War Navy technicians: mortality survey after 40 years". PMID- 12543631 TI - Exercise pressor reflex in decerebrate and anesthetized rats. AB - I investigated whether muscular contraction evokes cardiorespiratory increases (exercise pressor reflex) in alpha-chloralose- and chloral hydrate-anesthetized and precollicular, midcollicular, and postcollicular decerebrated rats. Mean arterial pressure (MAP), heart rate (HR), and minute ventilation (Ve) were recorded before and during 1-min sciatic nerve stimulation, which induced static contraction of the triceps surae muscles, and during 1-min stretch of the calcaneal tendon, which selectively stimulated mechanosensitive receptors in the muscles. Anesthetized rats showed various patterns of MAP response to both stimuli, i.e., biphasic, depressor, pressor, and no response. Sciatic nerve stimulation to muscle in precollicular decerebrated rats always evoked spontaneous running, so the exercise pressor reflex was not determined from these preparations. None of the postcollicular decerebrated rats showed a MAP response or spontaneous running. Midcollicular decerebrated rats consistently showed biphasic blood pressure response to both stimulations. The increases in MAP, HR, and Ve were related to the tension developed. The static contractions in midcollicular decerebrated rats (381 +/- 65 g developed tension) significantly increased MAP, HR, and Ve from 103 +/- 12 to 119 +/- 24 mmHg, from 386 +/- 30 to 406 +/- 83 beats/min, and from 122 +/- 7 to 133 +/- 25 ml/min, respectively. After paralysis, sciatic nerve stimulation had no effect on MAP, HR, or Ve. These results indicate that the midcollicular decerebrated rat can be a model for the study of the exercise pressor reflex. PMID- 12543632 TI - Tonin in rat heart with experimental hypertrophy. AB - The present study was undertaken to determine tonin expression and activity in rat heart presenting isoproterenol-induced hypertrophy. Renin, angiotensin converting enzyme (ACE), and angiotensinogen (AG) expression were also determined. Wistar rats were treated with isoproterenol for 7 days (5 mg x kg(-1) x day(-1) sc). For untreated animals, the levels of tonin-specific activity in the atrium were 2.6- and 5.5-fold higher than those of the left and right ventricle, respectively. After treatment, the levels of tonin-specific activity increased twofold in the atrium but did not change in the ventricles. Renin expression was not detectable in these structures, and ACE expression levels did not change with treatment. AG expression was detected in the left ventricle at very low levels compared with the atrium and increased significantly only in the hypertrophied atrium (1.8-fold). Tonin mRNA was not detected in the ventricle but was found at low levels in the atrium, which increased after isoproterenol treatment. Our results permit us to conclude that tonin may play a role in the process of heart hypertrophy in the rat. PMID- 12543633 TI - Uniaxial strain upregulates matrix-degrading enzymes produced by human vascular smooth muscle cells. AB - Arteries remodel in response to environmental changes. We investigated whether mechanical strain modulates production of matrix metalloproteinase (MMP)-2 and -9 by cultured vascular smooth muscle cells (SMC). MMP-2 and MMP-9 expression were tested using human saphenous vein SMC cultured on silicone membranes at rest or subjected to physiological levels (5%) of stationary or cyclical (1 Hz) uniaxial strain. Compared with control, stationary strain significantly increased MMP-2 mRNA levels at all time points, whereas cyclic strain decreased it after 48 h. Both secreted and cell-associated pro-MMP-2 levels were increased by stationary strain at all times (P < 0.01), whereas cyclic strain decreased secreted levels after 48 h (P < 0.02). MMP-9 mRNA levels and pro-MMP-9 protein were increased after 48 h of stationary stretch (P < 0.01) compared with both no strain and cyclic strain. Our study indicates that vascular SMC show a selective response to different types of strain. We suggest that local increases in stationary mechanical strain resulting from stenting, hypertension, or atherosclerosis may lead to enhanced matrix degradation by SMC. PMID- 12543634 TI - Angiogenic growth factor expression in rat skeletal muscle in response to exercise training. AB - Angiogenesis occurs in skeletal muscle in response to exercise training. To gain insight into the regulation of this process, we evaluated the mRNA expression of factors implicated in angiogenesis over the course of a training program. We studied sedentary control (n = 17) rats and both sedentary (n = 18) and exercise trained (n = 48) rats with bilateral femoral artery ligation. Training consisted of treadmill exercise (4 times/day, 1-24 days). Basal mRNA expression in sedentary control muscle was inversely related to muscle vascularity. Angiogenesis was histologically evident in trained white gastrocnemius muscle by day 12. Training produced initial three- to sixfold increases in VEGF, VEGF receptors (KDR and Flt), the angiopoietin receptor (Tie-2), and endothelial nitric oxide synthase mRNA, which dissipated before the increase in capillarity, and a substantial (30- to 50-fold) but transient upregulation of monocyte chemoattractant protein 1 mRNA. These results emphasize the importance of early events in regulating angiogenesis. However, we observed a sustained elevation of the angiopoietin 2-to-angiopoietin 1 ratio, suggesting continued vascular destabilization. The response to exercise was (in general) tempered in high oxidative muscles. These findings place importance on cellular events coupled to the onset of angiogenesis. PMID- 12543635 TI - Role of kappa-opioid receptor activation in pharmacological preconditioning of swine. AB - Pharmacological preconditioning with kappa-opioid receptor agonists is proarrhythmic and exerts antipreconditioning effects in rats. In swine, it is unknown whether kappa-opioid receptor stimulation plays a role in pharmacological preconditioning. Swine were preconditioned with 1) saline (controls), 2) [d Ala(2),d-Leu(5)]enkephalin (DADLE), 3) morphine, 4) pentazocine, 5) norbinaltorphimine (nor-BNI), 6) DADLE + nor-BNI, 7) morphine + nor-BNI, or 8) pentazocine + nor-BNI before occlusion (45 min) and reperfusion (180 min) of the left anterior descending coronary artery. Infarct size to area at risk (IS), regional (systolic shortening) and global (pressures and flows) myocardial function, and arrhythmia occurrence were assessed. Only DADLE + nor-BNI preconditioning significantly decreased infarct size compared with controls (47 +/- 13 vs. 65 +/- 5%, P < 0.05); morphine preconditioning was not cardioprotective with or without kappa-opioid receptor blockade (nor-BNI). DADLE preconditioning significantly increased ischemia-induced arrhythmias relative to controls, whereas pentazocine-preconditioned animals (n = 2) experienced intractable ventricular fibrillation during ischemia. kappa-Opioid receptor blockade with DADLE or pentazocine preconditioning alleviated proarrhythmic effects. These results suggest that kappa-opioid receptor activation during pharmacological preconditioning is proarrhythmic in swine. PMID- 12543636 TI - Desipramine attenuates loss of cardiac sympathetic neurotransmitters produced by congestive heart failure and NE infusion. AB - We reported recently that inhibition of neuronal reuptake of norepinephrine (NE) by desipramine prevented the reduction of sympathetic neurotransmitters in the failing right ventricle of right heart failure animals. In this study, we studied whether desipramine also reduced the sympathetic neurotransmitter loss in animals with left heart failure induced by rapid ventricular pacing (225 beats/min) or after chronic NE infusion (0.5 microg. kg(-1). min(-1)). Desipramine was given to the animals for 8 wk beginning with rapid ventricular pacing or NE infusion. Animals receiving no desipramine were studied as controls. We measured myocardial NE content, NE uptake activity, and sympathetic NE, tyrosine hydroxylase, and neuropeptide Y profiles by histofluorescence and immunocytochemical techniques. Effects of desipramine on NE uptake inhibition were evidenced by potentiation of the pressor response to exogenous NE and reduction of myocardial NE uptake activity. Desipramine treatment had no effect in sham or saline control animals but attenuated the reduction of sympathetic neurotransmitter profiles in the left ventricles of animals with rapid cardiac pacing and NE infusion. In contrast, the panneuronal marker protein gene product 9.5 profile was not affected by either rapid pacing or NE infusion, nor was it changed by desipramine treatment in the heart failure animals. The study confirms that excess NE contributes to the reduction of cardiac sympathetic neurotransmitters in heart failure. In addition, it shows that the anatomic integrity of the sympathetic nerves is relatively intact and that the neuronal damaging effect of NE involves the uptake of NE or its metabolites into the sympathetic nerves. PMID- 12543637 TI - Respiratory sinus arrhythmia is associated with efficiency of pulmonary gas exchange in healthy humans. AB - Respiratory sinus arrhythmia (RSA) may be associated with improved efficiency of pulmonary gas exchange by matching ventilation to perfusion within each respiratory cycle. Respiration rate, tidal volume, minute ventilation (.VE), exhaled carbon dioxide (.VCO(2)), oxygen consumption (.VO(2)), and heart rate were measured in 10 healthy human volunteers during paced breathing to test the hypothesis that RSA contributes to pulmonary gas exchange efficiency. Cross spectral analysis of heart rate and respiration was computed to calculate RSA and the coherence and phase between these variables. Pulmonary gas exchange efficiency was measured as the average ventilatory equivalent of CO(2) (.VE/.VCO(2)) and O(2) (.VE/.VO(2)). Across subjects and paced breathing periods, RSA was significantly associated with CO(2) (partial r = -0.53, P = 0.002) and O(2) (partial r = -0.49, P = 0.005) exchange efficiency after controlling for the effects of age, respiration rate, tidal volume, and average heart rate. Phase between heart rate and respiration was significantly associated with CO(2) exchange efficiency (partial r = 0.40, P = 0.03). These results are consistent with previous studies and further support the theory that RSA may improve the efficiency of pulmonary gas exchange. PMID- 12543638 TI - Adenosine receptors mediate glutamate-evoked arteriolar dilation in the rat cerebral cortex. AB - We tested the hypothesis that adenosine (Ado) mediates glutamate-induced vasodilation in the cerebral cortex by monitoring pial arteriole diameter in chloralose-anesthetized rats equipped with closed cranial windows. Topical application of 100 microM glutamate and 100 microM N-methyl-d-aspartate (NMDA) dilated pial arterioles (baseline diameter 25 +/- 2 microm) by 17 +/- 1% and 18 +/- 4%, respectively. Coapplication of the nonselective Ado receptor antagonist theophylline (Theo; 10 microM) significantly reduced glutamate- and NMDA-induced vasodilation to 4 +/- 2% (P < 0.01) and 6 +/- 2% (P < 0.05), whereas the Ado A(1) receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (0.1 microM) had no effect. Moreover, application of the Ado A(2A) receptor-selective antagonist 4-(2 [7-amino-2-(2-furyl)(1,2,4)triazolo(2,3-a)(1,3,5)triazin-5-ylamino]ethyl)phenol (ZM-241385), either by superfusion (0.1 microM, 1 microM) or intravenously (1 mg/kg), significantly inhibited the pial arteriole dilation response to glutamate. Neither Theo nor ZM-241385 affected vascular reactivity to mild hypercapnia induced by 5% CO(2) inhalation. These results suggest that Ado contributes to the dilation of rat cerebral arterioles induced by exogenous glutamate, and that the Ado A(2A) receptor subtype may be involved in this dilation response. PMID- 12543639 TI - Ischemia and reperfusion of skeletal muscle lead to the appearance of a stable lipid free radical in the circulation. AB - Both ischemia and reperfusion injury and contractile activity are associated with the generation of reactive oxygen species and free radicals by skeletal muscle. In addition, exercise has been reported to lead to the formation of a circulating free radical species that is detectable in the blood by spin trapping before analysis by electron-spin resonance (ESR) techniques. Previous analysis of the ESR signal indicated that the circulating species is either a carbon- or oxygen centered lipid-derived free radical. The current data indicate that this species is present in the blood of anesthetized rats after 4-h ischemia and 1 h of reperfusion of a single hindlimb. During 4 h of ischemia, the species was also present in microdialysates from the tibialis anterior muscle but was unchanged in magnitude compared with control tissue. During 1 h of reperfusion, the signal intensity increased by a mean of 420% (P < 0.05, n = 4). Hydroxyl radical activity in the interstitial fluid also significantly increased during ischemia and further increased by a mean of 210% (P < 0.05, n = 4) during reperfusion. No changes in interstitial superoxide levels were seen, but interstitial PGE(2) content also increased during reperfusion. A significant positive correlation was found between the magnitude of the ESR signal and both the hydroxyl radical activity and PGE(2) content of microdialysis fluids. These data support the hypothesis that the circulating free radical species is formed in the interstitial fluid by hydroxyl radical interaction with a lipid that may be released from reperfused tissue with a similar pattern to prostanoids. PMID- 12543640 TI - Syndecan-4 modulates basic fibroblast growth factor 2 signaling in vivo. AB - Syndecan-4 is one of the principal heparan sulfate-carrying proteins on the cell surface. Unlike other members of syndecan family, syndecan-4 mediates phosphatidylinositol 4,5-bisphosphate 2 (PIP(2))-dependent PKC-alpha activation, and overexpression of syndecan-4 in vitro results in enhanced FGF2 signaling. The present study was designed to test the functional effect of increased syndecan-4 expression in endothelial cells in transgenic mice. Several transgenic mice lines expressing syndecan-4 cDNA under control of human endothelial nitric oxide (NO) synthase (eNOS) promoter were generated. Exogenous syndecan-4 was mainly expressed in the heart, brain, and lungs. In particular, the heart demonstrated the greatest increase in the ratio of transgenic-to-native syndecan-4 gene expression. Vessels from the eNOS-syndecan-4 mice demonstrated more pronounced vasodilation to FGF2 but not to VEGF-A(165), sodium nitroprusside, and A 23187 compared with wild-type mice. To elucidate the mechanism of this effect, we measured NO release from primary cardiac endothelial cells isolated from transgenic or wild-type adult mice. Cells from the eNOS-syndecan-4 transgenic mice had a significant increase in FGF2- and VEGF-A(165)-induced NO release compared with endothelial cells from the wild-type mice. However, the absolute magnitude of this increase was higher for FGF2 than VEGF-A(165). In conclusion, enhanced syndecan-4 expression in mouse cardiac endothelial cells results in preferential augmentation of FGF2 but not VEGF-A(165)-induced NO release. PMID- 12543641 TI - E-Tmod capping of actin filaments at the slow-growing end is required to establish mouse embryonic circulation. AB - Tropomodulins are a family of proteins that cap the slow-growing end of actin filaments. Erythrocyte tropomodulin (E-Tmod) stabilizes short actin protofilaments in erythrocytes and caps longer sarcomeric actin filaments in striated muscles. We report the knockin of the beta-galactosidase gene (LacZ) under the control of the endogenous E-Tmod promoter and the knockout of E-Tmod in mouse embryonic stem cells. E-Tmod(-/-) embryos die around embryonic day 10 and exhibit a noncontractile heart tube with disorganized myofibrils and underdevelopment of the right ventricle, accumulation of mechanically weakened primitive erythroid cells in the yolk sac, and failure of primary capillary plexuses to remodel into vitelline vessels, all required to establish blood circulation between the yolk sac and the embryo proper. We propose a hemodynamic "plexus channel selection" mechanism as the basis for vitelline vascular remodeling. The defects in cardiac contractility, vitelline circulation, and hematopoiesis reflect an essential role for E-Tmod capping of the actin filaments in both assembly of cardiac sarcomeres and of the membrane skeleton in erythroid cells that is not compensated for by other proteins. PMID- 12543642 TI - Reactive oxygen species modulate coronary wall shear stress and endothelial function during hyperglycemia. AB - Hyperglycemia is associated with generation of reactive oxygen species (ROS), and this action may contribute to accelerated atherogenesis. We tested the hypothesis that hyperglycemia produces alterations in left anterior descending coronary artery (LAD) wall shear stress concomitant with endothelial dysfunction and ROS production in dogs (n = 12) instrumented for measurement of LAD blood flow, velocity, and diameter. Dogs were randomly assigned to receive vehicle (0.9% saline) or the superoxide dismutase mimetic 4- hydroxy-2,2,6,6 tetramethylpiperidine-1-oxyl (tempol) and were administered intravenous infusions of d-glucose to achieve target blood glucose concentrations of 350 and 600 mg/dl (moderate and severe hyperglycemia, respectively). Endothelial function and ROS generation were assessed by coronary blood flow responses to acetylcholine (10, 30, and 100 ng/kg) and dihydroethidium fluorescence of myocardial biopsies, respectively. Indexes of wall shear stress were calculated with conventional fluid dynamics theory. Hyperglycemia produced dose-related endothelial dysfunction, increases in ROS production, and reductions in oscillatory shear stress that were normalized by tempol. The results suggest a direct association between hyperglycemia-induced ROS production, endothelial dysfunction, and decreases in oscillatory shear stress in vivo. PMID- 12543643 TI - A glutamate mutase is involved in the biosynthesis of the lipopeptide antibiotic friulimicin in Actinoplanes friuliensis. AB - Actinoplanes friuliensis produces the lipopeptide antibiotic friulimicin. This antibiotic is active against gram-positive bacteria such as multiresistant Enterococcus and Staphylococcus strains. It consists of 10 amino acids that form a ring structure and 1 exocyclic amino acid to which an acyl residue is attached. By a reverse genetic approach, biosynthetic genes were identified that are required for the nonribosomal synthesis of the antibiotic. In close proximity two genes (glmA and glmB) were found which are involved in the production of methylaspartate, one of the amino acids of the peptide core. Methylaspartate is synthesized by a glutamate mutase mechanism, which was up to now only described for glutamate fermentation in Clostridium sp. or members of the family ENTEROBACTERIACEAE: The active enzyme consists of two subunits, and the corresponding genes overlap each other. To demonstrate enzyme activity in a heterologous host, it was necessary to genetically fuse glmA and glmB. The resulting gene was overexpressed in Streptomyces lividans, and the fusion protein was purified in an active form. For gene disruption mutagenesis, a host-vector system was established which enables genetic manipulation of Actinoplanes spp. for the first time. Thus, targeted inactivation of biosynthetic genes was possible, and their involvement in friulimicin biosynthesis was demonstrated. PMID- 12543644 TI - Biological properties of novel antistaphylococcal quinoline-indole agents. AB - The antibacterial properties of novel quinoline-indole (QI) agents were examined. QI agents demonstrated potent bactericidal activities against Staphylococcus aureus, killing by lytic and nonlytic mechanisms. S. aureus mutants resistant to a lytic QI agent (SEP 155342) and a nonlytic QI agent (SEP 118843) arose at frequencies of 1.4 x 10(-9) and 1.2 x 10(-8), respectively, by selection at four times the MICs. Mutants resistant to QI agent SEP 155342 were unstable, but mutants resistant to QI agent SEP 118843 displayed stable resistance. Mutants resistant to QI agent SEP 118843 were not cross resistant to other inhibitors, including QI agent SEP 155342. Addition of QI agents SEP 118843 and SEP 155342 at four times the MIC caused nonspecific inhibition of several macromolecular biosynthetic pathways in S. aureus. Within 10 min, QI agents SEP 118843 and SEP 155342 both interfered with bacterial membrane integrity, as measured by uptake of propidium iodide. Agents from the two classes of the QI agents probably kill staphylococci by separate mechanisms which, nevertheless, both involve interference with cytoplasmic membrane function. Precise structure-activity relationships for the division of QI agents into two classes could not be determined. However, lytic activity was often associated with substitution of a basic amine at position 4 of the quinoline nucleus, whereas compounds with nonlytic activity usually contained an aromatic ring with or without a methoxy substituent at position 4. Nonlytic QI agents such as SEP 118843 may possess selective activity against the prokaryotic membrane since this compound failed to lyse mouse erythrocytes when it was added at a concentration equivalent to four times the MIC for S. aureus. PMID- 12543645 TI - In vitro parasiticidal effect of Nitazoxanide against Echinococcus multilocularis metacestodes. AB - When humans serve as inadvertent intermediate hosts for Echinococcus multilocularis, disease (alveolar echinococcosis [AE]) may result from the expanding parasite metacestode in visceral organs, mostly in the liver. Benzimidazole carbamate derivatives such as mebendazole and albendazole are used for chemotherapeutic treatment of AE. However, these treatments are, in most cases, parasitistatic rather than parasiticidal. As treatment is discontinued, a recurrence of parasite growth has been observed in many AE patients with nonradical resections. The only curative treatment for AE is radical surgical resection of the parasite tissue and support by chemotherapy. As there is a need for new treatment options for AE, the in vitro efficacy of nitazoxanide (NTZ), a broad-spectrum drug used against intestinal parasites and bacteria, was investigated. We showed that in vitro treatment of E. multilocularis metacestodes with NTZ induced high levels of alkaline phosphatase activity in the medium. Concurrently, distinct morphological and ultrastructural alterations were detected. Most significantly, two distinct types of alterations were observed as soon as after 3 h of NTZ treatment. At first, the drug induced a peripheral output of membranous vesicles from the tegumental membrane into the laminated layer. Simultaneously, germinal layer-associated undifferentiated cells produced large vacuoles filled with lipid-like and often electron-dense membranous segments. Other alterations were observed at later time points, including vacuolization of the germinal layer, accumulation of lipid droplets, and lastly, loss of microtriches and separation of the laminated and germinal layers. The pattern of damage induced by NTZ was different from the alterations earlier observed in albendazole sulfoxide-treated vesicles. The nonviability of NTZ treated metacestodes was confirmed through bioassay, i.e., inoculation of treated and untreated parasites into mice. These experiments demonstrate the in vitro parasiticidal effect of NTZ on E. multilocularis metacestodes. PMID- 12543646 TI - Trypanosoma cruzi inactivation in human platelet concentrates and plasma by a psoralen (amotosalen HCl) and long-wavelength UV. AB - Trypanosoma cruzi, the protozoan pathogen that causes Chagas' disease, can be found in the blood of infected individuals for their entire life span. This presents a serious challenge in safeguarding blood products. Transmission of T. cruzi from blood products is a frequent occurrence in Latin America, where Chagas' disease is endemic. This study was designed to determine whether T. cruzi could be inactivated in human platelet concentrates and plasma by a photochemical treatment process with long-wavelength UV A light (UVA, 320 to 400 nm) plus the psoralen amotosalen HCl (Cerus Corporation). Units of platelet concentrates (300 ml) and plasma (300 ml) were intentionally contaminated with approximately 10(6) T. cruzi trypomastigotes, the T. cruzi form found in the bloodstream, per ml. The viability of T. cruzi after photochemical inactivation was determined by their ability to replicate in 3T3 fibroblasts. Controls, including treatment with 150 micro M amotosalen or 3 J/cm(2) UVA alone, did not lead to reduction of the viability of T. cruzi in plasma or platelet concentrates. However, treatment with 150 micro M amotosalen plus 3 J/cm(2) UVA inactivated T. cruzi to undetectable levels in plasma and platelet concentrates. This represented a >5.4-log reduction of T. cruzi in platelet concentrates and >5.0-log reduction of T. cruzi in plasma. We conclude that the amotosalen plus UVA photochemical inactivation technology is effective in inactivating high levels of protozoan pathogens, such as T. cruzi, in platelet concentrates and plasma, as has been previously shown for numerous viruses and bacteria. PMID- 12543647 TI - MC21-A, a bactericidal antibiotic produced by a new marine bacterium, Pseudoalteromonas phenolica sp. nov. O-BC30(T), against methicillin-resistant Staphylococcus aureus. AB - We previously reported a new marine bacterium, Pseudoalteromonas phenolica sp. nov. O-BC30(T), which produced a bactericidal antibiotic against methicillin resistant Staphylococcus aureus (MRSA). In the present study, we purified an anti MRSA substance (MC21-A) from the methanol extract of the cells of P. phenolica O BC30(T) and analyzed its chemical structure. MC21-A was determined to be 3,3',5,5'-tetrabromo-2,2'-biphenyldiol by spectrometric analyses. Its anti-MRSA activity against 10 clinical isolates of MRSA was comparable to that of vancomycin (MC21-A MICs, 1 to 2 micro g/ml; vancomycin MICs, <0.25 to 2 micro g/ml). This substance was also high active against Enterococcus serolicida, Enterococcus faecium, and Enterococcus faecalis but was less active against Streptococcus spp. A time-kill study also demonstrated that MC21-A was bactericidal and that its killing rate was much higher than that of vancomycin. The postantibiotic effect (PAE) of MC21-A against a clinical MRSA isolate, strain E 31243, was also comparable to that of vancomycin (MC21-A PAEs, 1.46 to 1.65 h; vancomycin PAEs, 0.84 to 1.43 h). However, a lysis experiment demonstrated that this substance failed to lyse MRSA cells. This substance also did not lyse human erythrocytes. A SYTOX Green staining experiment implied that this substance permeabilized the cell membrane of MRSA as its mode of action. When its activities against a hypersensitive Escherichia coli mutant (KO 1489) and wild type strains were tested, MC21-A exhibited higher levels of activity against the former. Furthermore, MC21-A was not cytotoxic to human normal fibroblast, rat pheochromocytoma, and Vero cells at concentrations up to 50 micro g/ml. These results suggest that MC21-A might be useful as a lead compound in the development of new types of anti-MRSA substances with modes of action different from those of vancomycin and teicoplanin. PMID- 12543648 TI - Macrolide-resistant Streptococcus pneumoniae and Streptococcus pyogenes in the pediatric population in Germany during 2000-2001. AB - In a nationwide study in Germany covering 13 clinical microbiology laboratories, a total of 307 Streptococcus pyogenes (mainly pharyngitis) and 333 Streptococcus pneumoniae (respiratory tract infections) strains were collected from outpatients less than 16 years of age. The MICs of penicillin G, amoxicillin, cefotaxime, erythromycin A, clindamycin, levofloxacin, and telithromycin were determined by the microdilution method. In S. pyogenes isolates, resistance rates were as follows: penicillin, 0%; erythromycin A, 13.7%; and levofloxacin, 0%. Telithromycin showed good activity against S. pyogenes isolates (MIC(90) = 0.25 micro g/ml; MIC range, 0.016 to 16 micro g/ml). Three strains were found to be telithromycin-resistant (MIC >/= 4 micro g/ml). Erythromycin-resistant strains were characterized for the underlying resistance genotype, with 40.5% having the efflux type mef(A), 38.1% having the erm(A), and 9.5% having the erm(B) genotypes. emm typing of macrolide-resistant S. pyogenes isolates showed emm types 4 (45.2%), 77 (26.2%), and 12 (11.9%) to be predominant. In S. pneumoniae, resistance rates were as follows: penicillin intermediate, 7.5%; penicillin resistant, 0%; erythromycin A, 17.4%; and levofloxacin, 0%. Telithromycin was highly active against pneumococcal isolates (MIC(90) 98%. Pretreating cells with NP-1 for 1 h prior to inoculation with untreated virus also prevents infection. NP-1, a cationic peptide, does not compete with viral envelope glycoproteins for binding to cellular heparan sulfate receptors, but it prevents viral entry. No VP16, a major viral tegument protein, is transported to the cell nucleus in the presence of NP-1. Infectious center assays demonstrate that NP-1 also inhibits cell-to-cell viral spread. Thus, NP-1 prevents virally mediated fusion events, entry, and cell-to-cell spread. This unique mechanism of anti-HSV activity, coupled with established antibacterial and possible anti-human immunodeficiency virus type 1 activities of defensins, render this family of compounds excellent candidates for further development as topical microbicides. PMID- 12543650 TI - Toward the development of a virus-cell-based assay for the discovery of novel compounds against human immunodeficiency virus type 1. AB - The emergence of human immunodeficiency virus type 1 (HIV-1) strains resistant to highly active antiretroviral therapy necessitates continued drug discovery for the treatment of HIV-1 infection. Most current drug discovery strategies focus upon a single aspect of HIV-1 replication. A virus-cell-based assay, which can be adapted to high-throughput screening, would allow the screening of multiple targets simultaneously. HIV-1-based vector systems mimic the HIV-1 life cycle without yielding replication-competent virus, making them potentially important tools for the development of safe, wide-ranging, rapid, and cost-effective assays amenable to high-throughput screening. Since replication of vector virus is typically restricted to a single cycle, a crucial question is whether such an assay provides the needed sensitivity to detect potential HIV-1 inhibitors. With a stable, inducible vector virus-producing cell line, the inhibitory effects of four reverse transcriptase inhibitors (zidovudine, stavudine, lamivudine, and didanosine) and one protease inhibitor (indinavir) were assessed. It was found that HIV-1 vector virus titer was inhibited in a single cycle of replication up to 300-fold without affecting cell viability, indicating that the assay provides the necessary sensitivity for identifying antiviral molecules. Thus, it seems likely that HIV-1-derived vector systems can be utilized in a novel fashion to facilitate the development of a safe, efficient method for screening compound libraries for anti-HIV-1 activity. PMID- 12543651 TI - Two mechanisms for human immunodeficiency virus type 1 inhibition by N-terminal modifications of RANTES. AB - C-C chemokine receptor 5 (CCR5) is the primary coreceptor for human immunodeficiency virus type 1 (HIV-1) infection. Native chemokines that bind to CCR5 inhibit HIV-1 infection, albeit weakly, but chemically modified chemokines inhibit infection more efficiently. We have investigated the inhibitory mechanism of three N-terminally modified RANTES variants (AOP-, NNY-, and PSC-RANTES) with the MT-2 human T-cell line stably expressing either native or mutated CCR5. The RANTES analogues showed the same rank order (PSC > NNY > AOP) in their capacity to induce prolonged CCR5 internalization, inhibit surface reexpression, and prevent HIV-1 infection on MT-2 cells expressing wild-type CCR5 or CCR5 with four C-terminal serine phosphorylation sites mutated to alanine. None of the RANTES analogues caused internalization of a C-terminal cytoplasmic domain deletion mutant of CCR5, and each derivative had equal potency in inhibiting HIV-1 infection of MT-2 cells expressing this mutant. We conclude that the C-terminal cytoplasmic residues of CCR5 are necessary for receptor sequestration by RANTES analogues but that the process and the relative activity of each derivative are not dependent upon phosphorylation of the C-terminal serine residues. Two mechanisms of antiviral activity are demonstrated: receptor blockade and receptor sequestration. Potency correlates with the ability to induce CCR5 sequestration but not with receptor binding, suggesting that sequestration may make the greater contribution to antiviral activity. PMID- 12543652 TI - Suboptimal antibiotic dosage as a risk factor for selection of penicillin resistant Streptococcus pneumoniae: in vitro kinetic model. AB - Optimizing pharmacokinetic/pharmacodynamic indices of antibiotics to obtain clinical and microbiological efficacy is essential, but dosing regimens must also be tailored to minimize the risk for emergence of resistance. The aim of the present study was to investigate whether certain concentrations of benzylpenicillin are critical for the selection of resistant subpopulations. A mixed culture of Streptococcus pneumoniae containing ca. 90% susceptible (MIC = 0.031 mg/liter), 9% intermediate (MIC = 0.25 mg/liter), and 1% resistant (MIC = 8 mg/liter) was studied in an in vitro kinetic model. The time that concentrations exceeded the MIC (T>MIC) for the three strains in the culture was varied by different initial concentrations of benzylpenicillin. Samples for viable counts were withdrawn at different times during 24 h and seeded on blood agar plates and on selective antibiotic-containing plates. The T>MIC varied from 46 to 100% for the susceptible strain, from 6 to 100% for the intermediate strain, and from 0 to 48% for the resistant strain. Our study, which may mimic the clinical situation with carriage of a mixed population of S. pneumoniae with different antibiotic susceptibilities, has shown that selection of resistant bacteria may easily occur if dosing regimens are only targeted toward fully susceptible strains. PMID- 12543653 TI - Activities of azithromycin and amphotericin B against Naegleria fowleri in vitro and in a mouse model of primary amebic meningoencephalitis. AB - Inhalation of fresh water containing the free-living ameba Naegleria fowleri may lead to a potentially fatal infection known as primary amebic meningoencephalitis. Amphotericin B is the only agent with established clinical efficacy in the treatment of primary amebic meningoencephalitis in humans, but therapy with this drug is often associated with adverse effects on the kidneys and other organs, and not all persons treated with amphotericin B have survived. We investigated the in vitro activity and in vivo efficacy of newer therapeutic agents in an attempt to identify other useful agents for treating primary amebic meningoencephalitis. Azithromycin has shown in vitro activity against Acanthamoeba spp. and in vivo activity against experimental toxoplasmosis. In our study, the MIC of azithromycin against N. fowleri was 13.4 micro M (10 micro g/ml), which was 123 times greater than the MIC of amphotericin B, which was 0.108 micro M (0.1 micro g/ml). Azithromycin protected 100% of mice infected with N. fowleri at a dose of 75 mg/kg/day for 5 days, whereas amphotericin B protected only 50% of mice at a dose of 7.5 mg/kg/day for 5 days, and all control mice died during the 28-day observation period. We conclude that azithromycin has both in vitro and in vivo activity versus N. fowleri and may be a useful addition to therapy for primary amebic meningoencephalitis. PMID- 12543654 TI - Activity of tigecycline (GAR-936), a novel glycylcycline, against Enterococci in the mouse peritonitis model. AB - A novel glycylcycline agent, tigecycline (GAR-936), was evaluated in vivo in the mouse model of peritonitis against three Enterococcus faecalis and four Enterococcus faecium isolates with different susceptibilities to vancomycin and tetracyclines, all of which were inhibited by 100 times the MIC) of antimicrobials. Furthermore, upon retest, these surviving organisms did not demonstrate increased MICs. Thus, this phenomenon does not reflect selection of antimicrobial-resistant mutants but rather survival of some organisms in high antimicrobial concentrations (heterotypic survival). An additional 44 clinical isolates of C. trachomatis from patients with single incident infections were tested against those from patients with recurrent or persistent infections, and heterotypic survival was seen in all isolates tested; hence, in vitro resistance did not correlate with the patient's apparent clinical outcome. PMID- 12543672 TI - MUC7 20-Mer: investigation of antimicrobial activity, secondary structure, and possible mechanism of antifungal action. AB - This study was aimed at examining the spectrum of antimicrobial activity of MUC7 20-mer (N-LAHQKPFIRKSYKCLHKRCR-C; residues 32 to 51 of MUC7, the low-molecular weight human salivary mucin, comprised of 357 residues) and comparing its antifungal properties to those of salivary histatin 5 (Hsn-5). We also examined the secondary structure of the 20-mer and the possible mechanism of its antifungal action. Our results showed that MUC7 20-mer displays potent killing activity against a variety of fungi and both gram-positive and gram-negative bacteria at micromolar concentrations. Time-dependent killing of Candida albicans and Cryptococcus neoformans by MUC7 20-mer and Hsn-5 indicated differences in killing rates between MUC7 20-mer and Hsn-5. The secondary structure prediction showed that MUC7 20-mer adopts an amphiphilic helix with distinguishable hydrophilic and hydrophobic faces (a characteristic that is associated with antimicrobial activity). In comparison to that of Hsn-5, the fungicidal activity of MUC7 20-mer against C. albicans seems to be independent of fungal cellular metabolic activity, as evidenced by its killing potency at a low temperature (4 degrees C) and in the presence of inhibitors of oxidative phosphorylation in the mitochondrial system. Fluorescence microscopy showed the ability of MUC7 20-mer to cross the fungal cell membrane and to accumulate inside the cells. The internalization of MUC7 20-mer was inhibited by divalent cations. Confocal microscopy of cells doubly labeled with MUC7 20-mer and a mitochondrion-specific dye indicated that mitochondria are not the target of MUC7 20-mer for either C. albicans or C. neoformans. PMID- 12543673 TI - Sterilizing activities of fluoroquinolones against rifampin-tolerant populations of Mycobacterium tuberculosis. AB - The bactericidal activities of ciprofloxacin, ofloxacin, levofloxacin, moxifloxacin, and gatifloxacin were tested in three models of rifampin-tolerant Mycobacterium tuberculosis persisters. Model 1 was a 100-day-old, unshaken, anaerobically adapted culture in which serial dilutions of the quinolones were incubated for 5 days and CFU counts were then done In models 2 and 3, 100 mg of rifampin/liter was added to the 100-day culture for 5 or 7 days to produce tolerant organisms that did not grow on plates; the rifampin was then washed off, fresh medium was added to allow recovery of growth on plates, and the culture was incubated for 7 days before CFU counts. In model 2, the quinolones were added after rifampin had been washed off, whereas in model 3 the quinolones were added to the cultures containing rifampin. In models 1 and 2, ciprofloxacin had the least bactericidal activity, ofloxacin and levofloxacin had greater activities, and moxifloxacin and gatifloxacin had the greatest activities. In model 3, ofloxacin had no detectable activity whereas moxifloxacin killed about log(10) 0.279 CFU of the persisters per ml at concentrations attainable in lesions; isoniazid had virtually no activity. These findings predict that ofloxacin will not be found to have effective sterilizing activity in clinical studies now planned whereas moxifloxacin will be able to shorten treatment. PMID- 12543674 TI - RWJ-54428 (MC-02,479), a new cephalosporin with high affinity for penicillin binding proteins, including PBP 2a, and stability to staphylococcal beta lactamases. AB - RWJ-54428 (MC-02,479) is a new cephalosporin active against gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). The potency of this new cephalosporin against MRSA is related to a high affinity for penicillin-binding protein 2a (PBP 2a), as assessed in a competition assay using biotinylated ampicillin as the reporter molecule. RWJ-54428 had high activity against MRSA strains COL and 67-0 (MIC of 1 micro g/ml) and also showed affinity for PBP 2a, with a 50% inhibitory concentration (IC(50)) of 0.7 micro g/ml. RWJ 54428 also displayed excellent affinity for PBP 5 from Enterococcus hirae R40, with an IC(50) of 0.8 micro g/ml and a MIC of 0.5 micro g/ml. The affinity of RWJ 54428 for PBPs of beta-lactam-susceptible S. aureus (MSSA), enterococci (E. hirae), and Streptococcus pneumoniae showed that the good affinity of RWJ-54428 for MRSA PBP 2a and E. hirae PBP 5 does not compromise its binding to susceptible PBPs. RWJ-54428 showed stability to hydrolysis by purified type A beta-lactamase isolated from S. aureus PC1. In addition, RWJ-54428 displayed low MICs against strains of S. aureus bearing the four classes of staphylococcal beta-lactamases, including beta-lactamase hyperproducers. The frequency of isolation of resistant mutants to RWJ-54428 from MRSA strains was very low. In summary, RWJ-54428 has high affinity to multiple PBPs and is stable to beta-lactamase, properties that may explain our inability to find resistance by standard methods. These data are consistent with its excellent activity against beta-lactam-resistant gram positive bacteria. PMID- 12543675 TI - AcrAB multidrug efflux pump is associated with reduced levels of susceptibility to tigecycline (GAR-936) in Proteus mirabilis. AB - Tigecycline has good broad-spectrum activity against many gram-positive and gram negative pathogens with the notable exception of the PROTEEAE: A study was performed to identify the mechanism responsible for the reduced susceptibility to tigecycline in Proteus mirabilis. Two independent transposon insertion mutants of P. mirabilis that had 16-fold-increased susceptibility to tigecycline were mapped to the acrB gene homolog of the Escherichia coli AcrRAB efflux system. Wild-type levels of decreased susceptibility to tigecycline were restored to the insertion mutants by complementation with a clone containing a PCR-derived fragment from the parental wild-type acrRAB efflux gene cluster. The AcrAB transport system appears to be associated with the intrinsic reduced susceptibility to tigecycline in P. mirabilis. PMID- 12543676 TI - Isoniazid activation defects in recombinant Mycobacterium tuberculosis catalase peroxidase (KatG) mutants evident in InhA inhibitor production. AB - Mycobacterium tuberculosis KatG catalyzes the activation of the antitubercular agent isoniazid to yield an inhibitor targeting enoyl reductase (InhA). However, no firm biochemical link between many KatG variants and isoniazid resistance has been established. In the present study, six distinct KatG variants identified in clinical Mycobacterium tuberculosis isolates resistant to isoniazid were generated by site-directed mutagenesis, and the recombinant mutant proteins (KatG(A110V), KatG(A139P), KatG(S315N), KatG(L619P), KatG(L634F), and KatG(D735A)) were purified and characterized with respect to their catalase peroxidase activities (in terms of k(cat)/K(m)), rates of free-radical formation from isoniazid oxidation, and, moreover, abilities to activate isoniazid. The A110V amino acid replacement did not result in significant alteration of KatG activities except that the peroxidase activity was enhanced. The other mutations, however, resulted in modestly reduced catalase and peroxidase catalytic efficiencies and, for the four mutants tested, significantly lower activities to oxidize isoniazid. Compared to the wild-type enzyme, the ability of the KatG(L634F), KatG(A139P), and KatG(D735A) variants to activate isoniazid decreased by 36%, 76%, and 73%, respectively, whereas the KatG(S315N) and KatG(L619P) variants completely lost their abilities to convert isoniazid into the InhA inhibitor. In addition, the inclusion of exogenous Mn(2+) to the isoniazid activation reaction mix significantly improved the ability of wild-type and KatG mutants to produce the InhA inhibitor. PMID- 12543677 TI - Genome-wide screening of Saccharomyces cerevisiae to identify genes required for antibiotic insusceptibility of eukaryotes. AB - The adverse reactions provoked by many antibiotics in humans are well documented but are generally poorly understood at the molecular level. To elucidate potential genetic defects that could give rise to susceptibility to prokaryote specific antibiotics in eukaryotes, we undertook genome-wide screens using the yeast Saccharomyces cerevisiae as a model of eukaryotes; our previous work with a small number of yeast mutants revealed some specific gene functions required for oxytetracycline resistance. Here, the complete yeast deletion strain collection was tested for growth in the presence of a range of antibiotics. The sensitivities of mutants revealed by these screens were validated in independent tests. None of the approximately 4,800 defined deletion strains tested were found to be sensitive to amoxicillin, penicillin G, rifampin, or vancomycin. However, two of the yeast mutants were tetracycline sensitive and four were oxytetracycline sensitive; encompassed among the latter were mutants carrying deletions in the same genes that we had characterized previously. Seventeen deletion strains were found to exhibit growth defects in the presence of gentamicin, with MICs for the strains being as low as 32 micro g ml(-1) (the wild type exhibited no growth defects at any gentamicin concentration tested up to 512 micro g ml(-1)). Strikingly, 11 of the strains that were most sensitive to gentamicin carried deletions in genes whose products are all involved in various aspects of vacuolar and Golgi complex (or endoplasmic reticulum) function. Therefore, these and analogous organelles, which are also the principal sites of gentamicin localization in human cells, appear to be essential for normal resistance to gentamicin in eukaryotes. The approach and data described here offer a new route to gaining insight into the potential genetic bases of antibiotic insusceptibilities in eukaryotes. PMID- 12543678 TI - Tandem amplification of a 28-kilobase region from the Yersinia enterocolitica chromosome containing the blaA gene. AB - Most Yersinia enterocolitica strains are resistant to beta-lactam antibiotics due to the production of one or two chromosomally encoded beta-lactamases. Strain Y56 is a Y. enterocolitica O:3 serotype natural isolate that is resistant to moderate amounts of penicillins and that produces a single class A beta-lactamase. To select mutants with increased levels of resistance to beta-lactam antibiotics, strain Y56 was grown on plates containing increasing amounts of ampicillin, and variants resistant to up to 500 micro g of ampicillin per ml were obtained. Chromosomal DNA from hyperresistant isolates was analyzed by Southern hybridization with a blaA-specific probe to detect gene rearrangements. The use of pulsed-field gel electrophoresis revealed that the increase in the resistance level correlated with the amplification in tandem of a DNA fragment of about 28 kb containing the blaA gene. The phenotype of these isolates was not stable, and they recovered the basal low resistance level when the ampicillin used for selection was withdrawn from the growth medium. This loss of resistance was followed by the recovery of the original chromosomal structure. To understand this amplification process, the 28-kb amplification unit was cloned, and the ends were sequenced. The analysis of these sequences did not reveal the presence of either repeats or transposable elements to explain this process. However, we found short sequences similar to some DNA gyrase target sequences that have been described. In addition, we observed that the frequency of appearance of ampicillin-hyperresistant isolates by amplification of the blaA locus was lowered in the presence of the gyrase inhibitor novobiocin. These findings suggest that the DNA gyrase could be involved in this amplification event. PMID- 12543679 TI - Effects of organic anion, organic cation, and dipeptide transport inhibitors on cefdinir in the isolated perfused rat kidney. AB - Cefdinir (Omnicef; Abbott Laboratories) is a cephalosporin antibiotic primarily eliminated by the kidney. Nonlinear renal elimination of cefdinir has been previously reported. Cefdinir renal transport mechanisms were studied in the erythrocyte-free isolated perfused rat kidney. Studies were performed with drug free perfusate and perfusate containing cefdinir alone to establish the baseline physiology and investigate cefdinir renal elimination characteristics. To investigate cefdinir renal transport mechanisms, inhibition studies were conducted by coperfusing cefdinir with inhibitors of the renal organic anion (probenecid), organic cation (tetraethylammonium), or dipeptide (glycylsarcosine) transport system. Cefdinir concentrations in biological samples were determined using reversed-phase high-performance liquid chromatography. Differences between treatments and controls were evaluated using analysis of variance and Dunnett's test. The excretion ratio (ER; the renal clearance corrected for the fraction unbound and glomerular filtration rate) for cefdinir was 5.94, a value indicating net renal tubular secretion. Anionic, cationic, and dipeptide transport inhibitors all significantly affected the cefdinir ER. With probenecid, the ER was reduced to 0.59, clearly demonstrating a significant reabsorptive component to cefdinir renal disposition. This finding was confirmed by glycylsarcosine studies, in which the ER was elevated to 7.95, indicating that reabsorption was mediated, at least in part, by the dipeptide transporter system. The effects of the organic cation tetraethylammonium, in which the ER was elevated to 7.53, were likely secondary in nature. The anionic secretory pathway was found to be the predominant mechanism for cefdinir renal excretion. PMID- 12543680 TI - Salmonella enterica serovar Typhimurium bla(PER-1)-carrying plasmid pSTI1 encodes an extended-spectrum aminoglycoside 6'-N-acetyltransferase of type Ib. AB - We have studied the aminoglycoside resistance gene, which confers high levels of resistance to both amikacin and gentamicin, that is carried by plasmid pSTI1 in the PER-1 beta-lactamase-producing strain of Salmonella enterica serovar Typhimurium previously isolated in Turkey. This gene, called aac(6')-Ib(11), was found in a class 1 integron and codes for a protein of 188 amino acids, a fusion product between the N-terminal moiety (8 amino acids) of the signal peptide of the beta-lactamase OXA-1 and the acetyltransferase. The gene lacked a plausible Shine-Dalgarno (SD) sequence and was located 45 nucleotides downstream from a small open reading frame, ORF-18, with a coding capacity of 18 amino acids and a properly spaced SD sequence likely to direct the initiation of aac(6')-Ib(11) translation. AAC(6')-Ib(11) had Leu118 and Ser119 as opposed to Gln and Leu or Gln and Ser, respectively, which were observed in all previously described enzymes of this type. We have evaluated the effect of Leu or Gln at position 118 by site-directed mutagenesis of aac(6')-Ib(11) and two other acetyltransferase gene variants, aac(6')-Ib(7) and -Ib(8), which naturally encode Gln118. Our results show that the combination of Leu118 and Ser119 confers an extended spectrum aminoglycoside resistance, with the MICs of all aminoglycosides in clinical use, including gentamicin, being two to eight times higher for strains with Leu118 and Ser119 than for those with Gln118 and Ser119. PMID- 12543681 TI - Efflux pump Lde is associated with fluoroquinolone resistance in Listeria monocytogenes. AB - Five Listeria monocytogenes isolates (CLIP 21369, CLIP 73298, CLIP 74811, CLIP 75679, and CLIP 79372) were found to be resistant to fluoroquinolones during the screening for antibiotic resistance of 488 L. monocytogenes isolates from human cases of listeriosis in France. On the basis of a fourfold or greater decrease in the ciprofloxacin MIC in the presence of reserpine, fluoroquinolone resistance was attributed to active efflux of the drugs. The lde gene (Listeria drug efflux; formerly lmo2741) encodes a 12-transmembrane-segment putative efflux pump belonging to the major facilitator superfamily of secondary transporters that displayed 44% identity with PmrA from Streptococcus pneumoniae. Insertional inactivation of the lde gene in CLIP 21369 indicated that the corresponding protein was responsible for fluoroquinolone resistance and was involved in the level of susceptibility to dyes such as ethidium bromide and acridine orange. PMID- 12543682 TI - Genetics of mefloquine resistance in the rodent malaria parasite Plasmodium chabaudi. AB - The genetic determinants of resistance to mefloquine in malaria parasites are unclear. Some studies have implied that amplification of, or mutations in, the multidrug resistance gene pfmdr1 in Plasmodium falciparum may be involved. Using the rodent malaria model Plasmodium chabaudi, we investigated the role of the orthologue of this gene, pcmdr1, in a stable mefloquine-resistant mutant, AS(15MF/3), selected from a sensitive clone. pcmdr1 exists as a single copy gene on chromosome 12 of the sensitive clone. In AS(15MF/3), the gene was found to have undergone duplication, with one copy translocating to chromosome 4. mRNA levels of pcmdr1 were higher in the mutant than in the parent sensitive clone. A partial genetic map of the translocation showed that other genes in addition to pcmdr1 had been cotranslocated. The sequences of both copies of pcmdr1 of AS(15MF/3) were identical to that of the parent sensitive clone. A cross was made between AS(15MF/3) and an unrelated mefloquine-sensitive clone, AJ. Phenotypic and molecular analysis of progeny clones showed that duplication and overexpression of the pcmdr1 gene was an important determinant of resistance. However, not all mefloquine-resistant progeny contained the duplicated gene, showing that at least one other gene was involved in resistance. PMID- 12543683 TI - Phenothiazines and thioxanthenes inhibit multidrug efflux pump activity in Staphylococcus aureus. AB - Efflux-related multidrug resistance (MDR) is a significant means by which bacteria can evade the effects of selected antimicrobial agents. Genome sequencing data suggest that Staphylococcus aureus may possess numerous chromosomally encoded MDR efflux pumps, most of which have not been characterized. Inhibition of these pumps, which may restore clinically relevant activity of antimicrobial agents that are substrates for them, may be an effective alternative to the search for new antimicrobial agents that are not substrates. The inhibitory effects of selected phenothiazines and two geometric stereoisomers of the thioxanthene flupentixol were studied using strains of S. aureus possessing unique efflux-related MDR phenotypes. These compounds had some intrinsic antimicrobial activity and, when combined with common MDR efflux pump substrates, resulted in additive or synergistic interactions. For S. aureus SA 1199B, which overexpresses the NorA MDR efflux pump, and for two additional strains of S. aureus having non-NorA-mediated MDR phenotypes, the 50% inhibitory concentration (IC(50)) for ethidium efflux for all tested compounds was between 4 and 15% of their respective MICs. Transport of other substrates was less susceptible to inhibition; the prochlorperazine IC(50) for acriflavine and pyronin Y efflux by SA-1199B was more than 60% of its MIC. Prochlorperazine and trans(E)-flupentixol were found to reduce the proton motive force (PMF) of S. aureus by way of a reduction in the transmembrane potential. We conclude that the mechanism by which phenothiazines and thioxanthenes inhibit efflux by PMF dependent pumps is multifactorial and, because of the unbalanced effect of these compounds on the MICs and the efflux of different substrates, may involve an interaction with the pump itself and, to a lesser extent, a reduction in the transmembrane potential. PMID- 12543684 TI - Molecular epidemiology of the SRL pathogenicity island. AB - The Shigella resistance locus (SRL), which is carried on the SRL pathogenicity island (PAI) in Shigella flexneri 2a YSH6000, mediates resistance to the antibiotics streptomycin, ampicillin, chloramphenicol, and tetracycline. In the present study, we investigated the distribution and structural variation of the SRL and the SRL PAI in 71 Shigella isolates and 28 other enteric pathogens by PCR and Southern analysis. The SRL and SRL-related loci, although absent from the other enteric pathogens evaluated in this study, were found to be present in a number of Shigella isolates. SRL PAI markers were also present in the majority of strains carrying the SRL and SRL-related loci. PCR linkage studies with six of these strains demonstrated that the SRL is carried on elements similar in structure and organization to the YSH6000 SRL PAI, consistent with the hypothesis that the SRL PAI may be involved in the spread of multiple-antibiotic resistance in these strains. PMID- 12543685 TI - Fosmidomycin, a novel chemotherapeutic agent for malaria. AB - In previous studies, fosmidomycin has been shown to possess activity against Plasmodium falciparum in vitro and in the mouse model. It has a novel mode of action through inhibition of 1-deoxy-D-xylulose 5-phosphate reductoisomerase, an enzyme of the nonmevalonate pathway of isoprenoid biosynthesis, which is absent in humans. In this open-label, uncontrolled trial, the efficacy and safety of fosmidomycin, in an oral dose of 1,200 mg every 8 h for 7 days, were evaluated in the treatment of acute uncomplicated Plasmodium falciparum malaria in 20 adult subjects in Gabon and Thailand. Clinical assessments were performed and thick blood smears were evaluated every 8 h until parasite clearance and resolution of symptoms were achieved; assessments continued at weekly intervals thereafter for the duration of the 28-day followup period. All subjects were clinically and parasitologically cured on day 7 (primary end point). Parasite and fever clearance were rapid, with means of 44 and 41 h, respectively. On day 28, seven out of nine subjects (78%) were cured in Gabon and two out of nine subjects (22%) were cured in Thailand. The drug was well tolerated, although mild gastrointestinal side effects were recorded for five subjects. Analysis of hematological and biochemical parameters showed no clinically significant changes throughout the study. Fosmidomycin is an effective and safe antimalarial drug, although its use as a single agent is restricted by the occurrence of recrudescent infections. However, its role in combination therapy should be explored. PMID- 12543686 TI - Influence of macrolide susceptibility on efficacies of clarithromycin and azithromycin against Streptococcus pneumoniae in a murine lung infection model. AB - We evaluated the activities of clarithromycin and azithromycin against 19 isolates of Streptococcus pneumoniae using a neutropenic lung infection model. The isolates included five susceptible isolates (clarithromycin and azithromycin MICs, /=64 micro g/ml). Infected mice were administered either saline (control), clarithromycin (4, 40, or 200 mg/kg of body weight twice daily or 200 mg/kg once daily), or azithromycin (4, 40, or 200 mg/kg once daily or 40 mg/kg twice daily) by oral gavage for 72 h. Mortality was assessed at regular intervals for 10 days, and survival in each group was compared to that of untreated controls. Animals infected with susceptible isolates demonstrated significant improvement in survival compared to the controls following treatment with either agent at doses of >/=40 mg/kg. In contrast, none of the regimens improved the survival of animals infected with isolates exhibiting high-level macrolide resistance. Among mice infected with strains expressing low-level resistance, significant improvement in survival compared to the controls was noted among isolates treated with clarithromycin at 40 (seven of nine isolates) and 200 (nine of nine isolates) mg/kg twice a day and with azithromycin at 40 (one of nine isolates) and 200 (three of nine isolates) mg/kg once a day. Animals infected with isolates of S. pneumoniae exhibiting low level, mefA-mediated macrolide resistance responded to treatment with clarithromycin at rates similar to those observed among mice infected with fully susceptible isolates. PMID- 12543687 TI - Lamivudine can exert a modest antiviral effect against human immunodeficiency virus type 1 containing the M184V mutation. AB - The M184V mutation in human immunodeficiency virus (HIV) reverse transcriptase is associated with high-level resistance to both (-)2',3'-dideoxy-3'-thiacytidine (3TC) and (-)2',3'-dideoxy-5-fluoro-3'-thiacytidine as well as low-level resistance to 2',3'-dideoxyinosine, 2',3'-dideoxycytidine, and abacavir. This mutation is also associated with diminished HIV replicative fitness as well as several functional changes in enzyme activity, including diminutions in polymerase processivity, pyrophosphorylysis, and nucleotide primer unblocking. Despite the fact that M184V encodes up to 1,000-fold resistance to 3TC, we asked whether this drug might still display some antiviral effect in regard to viruses containing this mutation. Cell-free assays revealed that high concentrations of 3TC triphosphate (i.e., >100 micro M) could affect chain termination and/or inhibit purified reverse transcriptase containing the M184V substitution. This effect became more pronounced with elongation of reverse transcriptase products. In newly infected cells (i.e., peripheral blood mononuclear cells), we found that the amount of full-length reverse transcriptase product was diminished in the presence of 2 to 10 micro M 3TC, although no decrease in the first product of the reverse transcriptase reaction, i.e., minus strong-stop DNA, was observed. In the presence of two other HIV inhibitors, e.g., nevirapine and indinavir, 3TC exerted additive effects in tissue culture at concentrations only marginally higher than the 50% inhibitory concentration (IC(50)). Reverse transcriptases cloned from clinical isolates harboring M184V in the context of multidrug resistance had similar IC(50) values for 3TC triphosphate compared to reverse transcriptase containing only the M184V mutation. These results suggest that viruses containing M184V can retain a higher degree of sensitivity to 3TC than previously assumed. PMID- 12543688 TI - Emergence in Klebsiella pneumoniae of a chromosome-encoded SHV beta-lactamase that compromises the efficacy of imipenem. AB - A Klebsiella pneumoniae isolate was identified that had reduced susceptibility to several expanded-spectrum cephalosporins and imipenem. That isolate produced a chromosome-encoded SHV-type beta-lactamase, SHV-38, that had an alanine to valine substitution in position Ambler 146 compared to beta-lactamase SHV-1. The kinetic parameters for purified beta-lactamases SHV-38 and SHV-1 showed that the hydrolytic spectrum of SHV-38 included only ceftazidime and imipenem. This report is the first example of an SHV-type beta-lactamase capable of hydrolyzing imipenem. PMID- 12543690 TI - In vitro and in vivo pharmacokinetics-pharmacodynamics of GV143253A, a novel trinem. AB - The aim of the present study was to characterize the pharmacokinetic pharmacodynamic relationship of GV143253A, a novel trinem anti-methicillin resistant Staphylococcus aureus (MRSA) agent active against gram-positive cocci, including multidrug-resistant clinical isolates. An in vitro pharmacodynamic study with methicillin-susceptible S. aureus (MSSA) and MRSA has shown that the duration of exposure to GV143253A rather than its concentration is the major determinant of the extent of bacterial killing. The in vitro findings were confirmed by use of a neutropenic murine model of thigh infection caused by MSSA ATCC 25923. From the dose-response curves, the static doses extrapolated for three different dosing intervals showed that more frequent dosing of GV143253A was more effective than less frequent dosing. A pharmacokinetic-pharmacodynamic analysis demonstrated that only the time during which the drug concentration exceeded the MIC (t>MIC) correlated with in vivo GV143253A activity. The value of t>MIC required to achieve a bacteriostatic effect in a thigh infection of neutropenic animals was 20% (95% confidence interval [CI], 18 to 22%) of the dosing interval. This result is similar to those reported in the literature for carbapenems and for GV104326A, another novel trinem compound. In addition, in order to compare the therapeutic efficacy of GV143253A to that of vancomycin in a thigh infection caused by MRSA in immunocompetent mice, suitable dosing regimens were designed on the basis of previous pharmacokinetic-pharmacodynamic findings for GV143253A and on the human pharmacokinetic profile of the glycopeptide. Although the pharmacokinetic profiles of the two agents were completely different, GV143253A showed good efficacy comparable to that of vancomycin, reducing by 4 log units the bacterial counts in the thighs of treated mice relative to untreated infected animals after 48 h of therapy. The results suggest that if the time of exposure to the pathogen above the MIC is at least 30% of the dosing interval, GV143253A could have a role in the clinical treatment of infections caused by MRSA, which is difficult to eradicate with current antibiotics. PMID- 12543689 TI - Selection of high-level resistance to human immunodeficiency virus type 1 protease inhibitors. AB - Protease inhibitors represent some of the most potent agents available for therapeutic strategies designed to inhibit human immunodeficiency virus type 1 (HIV-1) replication. Under certain circumstances the virus develops resistance to the inhibitor, thereby negating the benefits of this therapy. We have carried out selections for high-level resistance to each of three protease inhibitors (indinavir, ritonavir, and saquinavir) in cell culture. Mutations accumulated over most of the course of the increasing selective pressure. There was significant overlap in the identity of the mutations selected with the different inhibitors, and this gave rise to high levels of cross-resistance. Virus particles from the resistant variants all showed defects in processing at the NC/p1 protease cleavage site in Gag. Selections with pairs of inhibitors yielded similar patterns of resistance mutations. A virus that could replicate at near toxic levels of the three protease inhibitors combined was selected. The pro sequence of this virus was similar to that of the viruses that had been selected for high-level resistance to each of the drugs singly. Finally, a molecular clone carrying the eight most common resistance mutations seen in these selections was characterized. The sequence of this virus was relatively stable during selection for revertants in spite of displaying poor processing at the NC/p1 site and having significantly reduced fitness. These results reveal patterns of drug resistance that extend to near the limits of attainable selective pressure with these inhibitors and confirm the patterns of cross-resistance for these three inhibitors and the attenuation of virion protein processing and fitness that accompanies high-level resistance. PMID- 12543691 TI - Interferons, interferon inducers, and interferon-ribavirin in treatment of flavivirus-induced encephalitis in mice. AB - We evaluated the prophylactic and therapeutic efficacy of interferon alpha-2b, pegylated interferon alpha-2b, poly(I. C), and Ampligen against Modoc virus encephalitis in an animal model for flavivirus infections. All compounds significantly delayed virus-induced morbidity (paralysis) and mortality (due to progressive encephalitis). Viral load (as measured on day 7 postinfection) was significantly reduced by 80 to 100% in the serum, brain, and spleen in mice that had been treated with either interferon alpha-2b, pegylated interferon alpha-2b, poly(I. C), or Ampligen. We also studied whether a combination of interferon alpha-2b and ribavirin (presently the standard therapy for the treatment of infections with hepatitis C virus) would be more effective than treatment with interferon alone. However, ribavirin did not enhance the inhibitory effect of interferon therapy in this animal model for flavivirus infections. PMID- 12543692 TI - Rapid in vivo screening of experimental drugs for tuberculosis using gamma interferon gene-disrupted mice. AB - We have developed a rapid new in vivo method for screening experimental drugs for their activity against Mycobacterium tuberculosis by using the gamma interferon gene-disrupted (GKO) C57BL/6 mouse. Due to the rapid growth of the infection, statistical differences indicating positive efficacy of active compounds can be seen after only 8 days of treatment. To validate this model, several fluoroquinolones, including ciprofloxacin, levofloxacin, moxifloxacin, and gatifloxacin, were tested in parallel. PMID- 12543693 TI - Nonconjugative transposition of the vanB-containing Tn5382-like element in Enterococcus faecium. AB - The vanB2 operon encoding glycopeptide resistance is an integral part of the putative conjugative transposon Tn5382. Characterization of clinical glycopeptide resistant derivatives from an epidemic ampicillin-resistant Enterococcus faecium strain showed precise chromosomal or plasmid insertions of a vanB2-containing Tn5382-like element. Conjugative transposition of the Tn5382-like element was not demonstrated in retransfer studies. PMID- 12543694 TI - Clinical isolates of Enterobacteriaceae producing extended-spectrum beta lactamases: prevalence of CTX-M-3 at a hospital in China. AB - The prevalence of extended-spectrum beta-lactamase-producing strains was demonstrated in 5 of 44 (11.4%) Escherichia coli, 17 of 43 (39.5%) Klebsiella pneumoniae, 3 of 50 (6.0%) Enterobacter cloacae, and 2 of 25 (8.0%) Citrobacter freundii strains at a teaching hospital in China. Nineteen of these 27 strains expressed CTX-M-3 beta-lactamase (pI 8.6). A subset of the clinical isolates expressing the CTX-M-3 enzyme, tested by pulsed-field gel electrophoresis, revealed multiple clones. Five isolates expressed a novel enzyme, SHV-43 (pI 8.0), which had two substitutions (Leu113Phe and Thr149Ser) compared with SHV-1. PMID- 12543695 TI - Genetic basis of antibiotic resistance in Streptococcus agalactiae strains isolated in a French hospital. AB - The genetic basis of antibiotic resistance in 113 unrelated group B streptococci was studied by PCR. Ninety-four strains were resistant to tetracycline minocycline, and tet(M) was detected in 85% of these isolates. Seventeen erythromycin-resistant strains contained the erm(B), erm(TR), or mef(A) gene. Eleven strains exhibited high-level resistance to kanamycin due to the presence of the aphA3 gene; eight of these strains were also highly resistant to streptomycin; aad-6-related sequences were detected in seven strains. PMID- 12543696 TI - Conjugative plasmid conferring resistance to olaquindox. AB - A conjugative plasmid, pOLA52, conferring resistance to the antibiotic growth promoter olaquindox has been isolated from Escherichia coli from swine manure. It also confers resistance to ampicillin and chloramphenicol and has a high frequency of transfer between strains of E. coli. Plasmid-borne olaquindox resistance has not been demonstrated before. PMID- 12543697 TI - Growth-inhibitory effects of artesunate, pyrimethamine, and pamaquine against Babesia equi and Babesia caballi in in vitro cultures. AB - Three antimalarial drugs, artesunate, pyrimethamine, and pamaquine, were evaluated for their growth-inhibitory effects against Babesia equi and Babesia caballi in in vitro culture. B. equi was more resistant to pyrimethamine than B. caballi. B. equi was also found to be more sensitive to artesunate and pamaquine than B. caballi. Of the three compounds, pyrimethamine gave the most promise for in vivo effectiveness. PMID- 12543698 TI - Molecular epidemiology of penicillin-resistant and ciprofloxacin-resistant Streptococcus pneumoniae in Canada. AB - Eighty-nine penicillin- and ciprofloxacin-resistant Streptococcus pneumoniae isolates were evaluated by serotyping and pulsed-field gel electrophoresis. Although penicillin-resistant isolates demonstrated considerable homogeneity, resistance to ciprofloxacin did not correlate with a reduction in genotypic variability. These results suggest that, unlike that of penicillin resistance, the spread of S. pneumoniae ciprofloxacin resistance in Canada is currently not attributable to clonal dissemination. PMID- 12543699 TI - Daily variations in ceftriaxone pharmacokinetics in rats. AB - The aim of this study was to determine whether the time of day ceftriaxone was administered modified its pharmacokinetics. Ceftriaxone was given intraperitoneally at either 0400, 1000, 1600, and 2200 h to Sprague-Dawley rats synchronized under a light-dark cycle of 12 h of light and 12 h of dark. Pharmacokinetic parameters were analyzed for the presence of a 24-h rhythm. Results showed significant daily variations (P < 0.05) in ceftriaxone clearance, with the highest values during the dark phase. It is concluded that time dependent variations in ceftriaxone pharmacokinetics may affect the therapeutic efficacy of current once-daily dosing schedules. PMID- 12543700 TI - Efficacy of amphotericin B or itraconazole in a murine model of central nervous system Aspergillus infection. AB - Given the greater than 90% lethality of clinical central nervous system (CNS) aspergillosis despite current therapies, there is a need for an animal model to study therapeutic strategies. We previously established a model of CNS aspergillosis by intracerebral infection and report here the results of treatment with the two therapies with the greatest clinical experience, i.e., treatments with amphotericin B (AMB) and itraconazole (ITZ). Mice were given cyclophosphamide to produce pancytopenia. AMB was given intraperitoneally (i.p.; 3 mg/kg of body weight) or intravenously (i.v.; 0.8 mg/kg) once daily. ITZ in cyclodextrin was given by gavage once daily at a dose of 100 mg/kg or twice daily at 50 mg/kg. Treatments were started at day 1 postinfection and given for 10 days. At day 15, survivors were euthanatized. Ninety percent of the mice given no treatment died by day 6, and 100% died by day 10. Mice treated with AMB either i.p. or i.v. had 40% survival. Mice treated with ITZ either once or twice per day had a median survival time of 10 days, compared with 4 days for control animals, but a survival rate of only 10%. AMB and ITZ prolonged survival (P, <0.0001 to <0.05) compared with controls. Brains from surviving mice had CFU of Aspergillus fumigatus. This model can be used to compare newer antifungals and to study combination therapy or immunotherapy to find better therapeutic alternatives. PMID- 12543701 TI - Levofloxacin and ciprofloxacin in vitro activities against 4,003 clinical bacterial isolates collected in 24 Italian laboratories. AB - Levofloxacin showed comparable in vitro susceptibility to ciprofloxacin among Enterobacteriaceae, Pseudomonas aeruginosa, enterococci, and Staphylococcus aureus, while greater susceptibility was observed in Stenotrophomonas maltophilia and Staphylococcus epidermidis, mainly when oxacillin resistant. The susceptibility of Streptococcus pneumoniae to levofloxacin reached 99%. PMID- 12543702 TI - Pharmacokinetics of florfenicol in healthy pigs and in pigs experimentally infected with Actinobacillus pleuropneumoniae. AB - A comparative in vivo pharmacokinetic study of florfenicol was conducted in 18 crossbred pigs infected with Actinobacillus pleuropneumoniae following intravenous (i.v.), intramuscular (i.m.), or oral (p.o.) administration of a single dose of 20 mg/kg. The disease model was confirmed by clinical signs, X rays, pathohistologic examinations, and organism isolation. Florfenicol concentrations in plasma were determined by a validated high-performance liquid chromatography method with UV detection at a wavelength of 223 nm. Pharmacokinetic parameters were calculated by using the MCPKP software (Research Institute of Traditional Chinese Veterinary Medicine, Lanzhou, China). The disposition of florfenicol after a single i.v. bolus was described by a two compartment model with values for the half-life at alpha phase (t(1/2alpha)), the half-life at beta phase (t(1/2beta)), the area under the concentration-time curve (AUC(0- infinity )), and the volume of distribution at steady state (V(ss)) of 0.37 h, 2.91 h, 64.86 micro g. h/ml, and 1.2 liter/kg, respectively. The concentration-time data fitted the one-compartment (after i.m.) and two compartment (after p.o.) models with first-order absorption. The values for the maximum concentration of drug in serum (C(max)), t(1/2alpha), t(1/2beta), and bioavailability after i.m. and p.o. dosing were 4.00 and 8.11 micro g/ml, 0.12 and 3.91 h, 13.88 and 16.53 h, and 122.7 and 112.9%, respectively, for the two models. The study showed that florfenicol was absorbed quickly and completely, distributed widely, and eliminated slowly in the infected pigs, and there was no statistically significant difference between the pharmacokinetic profiles for the infected and healthy pigs. PMID- 12543703 TI - Serotype distribution and antimicrobial resistance of Streptococcus pneumoniae isolated in Algiers, Algeria. AB - There are few data on antibiotic resistance of Streptococcus pneumoniae in Algeria. Among 309 strains, 34.6% were penicillin G-nonsusceptible S. pneumoniae strains (25.2% were intermediate and 9.4% were resistant). Serotypes 1, 5, 14, and 6 were the most frequent in invasive child infections. A multicenter study to standardize the national guidelines is needed. PMID- 12543704 TI - Is it time to change fluoroquinolone breakpoints for Salmonella spp.? PMID- 12543705 TI - Reinterpretation of susceptibility data using current NCCLS breakpoint criteria. PMID- 12543706 TI - Volumetric properties of proteins. AB - Structural and thermodynamic characterizations of a variety of intra- and intermolecular interactions stabilizing/destabilizing protein systems represent a major part of multidisciplinary efforts aimed at solving the problems of protein folding and binding. To this end, volumetric techniques have been successfully used to gain insights into protein hydration and intraglobular packing. Despite the fact that the use of volumetric measurements in protein-related studies dates back to the 1950s, such measurements still represent a relatively untapped yet potentially informative means for tackling the problems of protein folding and binding. This notion has been further emphasized by recent advances in the development of highly sensitive volumetric instrumentation that has led to intensifying volumetric investigations of protein systems. This paper reviews the volumetric properties of proteins and their low-molecular-weight analogs, in particular, discussing the recent progress in the use of volumetric data for studying conformational transitions of proteins as well as protein-ligand, protein-protein, and protein-nucleic acid interactions. PMID- 12543707 TI - The state of lipid rafts: from model membranes to cells. AB - Lipid raft microdomains were conceived as part of a mechanism for the intracellular trafficking of lipids and lipid-anchored proteins. The raft hypothesis is based on the behavior of defined lipid mixtures in liposomes and other model membranes. Experiments in these well-characterized systems led to operational definitions for lipid rafts in cell membranes. These definitions, detergent solubility to define components of rafts, and sensitivity to cholesterol deprivation to define raft functions implicated sphingolipid- and cholesterol-rich lipid rafts in many cell functions. Despite extensive work, the basis for raft formation in cell membranes and the size of rafts and their stability are all uncertain. Recent work converges on very small rafts <10 nm in diameter that may enlarge and stabilize when their constituents are cross-linked. PMID- 12543708 TI - The enzymes, regulation, and genetics of bile acid synthesis. AB - The synthesis and excretion of bile acids comprise the major pathway of cholesterol catabolism in mammals. Synthesis provides a direct means of converting cholesterol, which is both hydrophobic and insoluble, into a water soluble and readily excreted molecule, the bile acid. The biosynthetic steps that accomplish this transformation also confer detergent properties to the bile acid, which are exploited by the body to facilitate the secretion of cholesterol from the liver. This role in the elimination of cholesterol is counterbalanced by the ability of bile acids to solubilize dietary cholesterol and essential nutrients and to promote their delivery to the liver. The synthesis of a full complement of bile acids requires 17 enzymes. The expression of selected enzymes in the pathway is tightly regulated by nuclear hormone receptors and other transcription factors, which ensure a constant supply of bile acids in an ever changing metabolic environment. Inherited mutations that impair bile acid synthesis cause a spectrum of human disease; this ranges from liver failure in early childhood to progressive neuropathy in adults. PMID- 12543709 TI - From the Lymphatic Research Foundation. PMID- 12543710 TI - The lymphatic continuum: the past, present, and exciting future of lymphatic research. PMID- 12543711 TI - The role of the National Center for Research Resources at the National Institutes of Health: infrastructure to forge a new road for lymphatic biology and therapeutics. AB - Lymphatic research has infrastructure needs ranging from the nursing support provided by General Clinical Research Centers to training grants for future clinician investigators. Both have high priority in the activities currently funded by the Division of Clinical Research at NCRR. Further into the future, the therapeutic development networks and embryonic stem cells resources that are currently being developed should seem equally to have been essential resources. PMID- 12543712 TI - An overview of the pathology and approaches to tissue engineering. AB - In tissue engineering, there is an attempt to culture living tissues for surgical transplantation. In vitro and in vivo approaches have produced vascular and cardiovascular components, cartilage, bone, intestine, and liver. Attempts to microdesign cell-culture support scaffolds have used a new generation of biocompatible and bioabsorbable polymers. Suspensions of donor cells are seeded onto protein-coated polymer scaffolds and grown to confluence in dynamic bioreactors. In vitro techniques produce monolayers of tissues. Denser masses are achieved by implanting monolayers onto a host, or by culturing cell/polymer constructs in vivo. Existing techniques have produced functioning heart valves from sheep endothelial cells and myofibroblasts. Cultured ovine arterial cells have replaced 2-cm segments of pulmonary artery in lambs. Chondrocyte cultures have produced a human-ear-shaped construct, temporo-mandibular joint discs, meniscal replacement devices, and human-phalange-shaped constructs, complete with a joint. The culture of composite tissue types has recently been reported. Intestinal organoid units containing a mesenchymal core with surrounding polarized epithelia have been used in lieu of an ileal pouch in Lewis rats, and the long-term culture of rat hepatocytes has revealed cellular differentiation and neomorphology resembling elements of a biliary drainage system. To sustain the in vitro culture of dense tissues prior to implantation, micro-electro mechanical systems (MEMS) fabrication technologies have been adapted to create wafers of polymer containing sealed, branching, vascular-type spaces. After seeding with rat lung endothelial cells, followed by 5 days of bioreactor culture, the result is an endothelial network with controlled blood flow rates, pressure, and hematocrit. When these customized vascular systems can be used to support in vitro culture, a new generation of dense, composite, morphologically complex tissues will be available for clinical development. PMID- 12543713 TI - Tissue engineering of the lymphatic system. AB - The field of tissue engineering has seen tremendous expansion in the last decade. In the last several years, tissue-engineering strategies to treat diseases of skin, cartilage, bone, bladder, blood vessel, tendon, and other tissues have been described. However, tissue-engineering approaches to treat diseases of the lymphatic system are currently nonexistent. We propose that acellular tissues, either native or engineered, could be exploited as a platform for the study of lymphatic biology, and for lymphatic tissue engineering. While speculative, this type of experimental model system could prove powerful for dissecting molecular and cellular events surrounding tumor invasion of lymphatics, as well as lymphangiogenesis. Scaffolds seeded with genetically engineered lymphatic cells could also be implanted to repopulate lymphatic vasculature. In the future, the lymphatic system will surely be added to the list of tissues and organs that prove amenable to tissue-engineering therapies. PMID- 12543715 TI - Research perspectives in inherited lymphatic disease. AB - The hereditary lymphedemas provide an opportunity to identify genes involved in normal and deranged lymphatic development. Genetic analysis of families with Milroy's disease identified mutations in VEGFR3 as a cause of congenital lymphedema, confirming the importance of VEGFC/VEGFR3 signaling in lymphatic development. These observations led to the identification of a mouse model for primary lymphedema, and subsequent analysis of this mouse model, using transgenic and gene transfer techniques, has provided initial clues to the development of a biologically based therapy for primary lymphedema. Of more importance from a public health perspective is the fact that manipulation of this pathway may lead to effective therapies for the more prevalent forms of secondary lymphedema. Identification of FOXC2 as the gene mutated in the lymphedema-distichiasis syndrome has revealed new molecular insight into lymphatic development. Molecular analysis of the FOXC2 pathway may provide clues to developmental pathways shared by the lymphatic system and the other developmental abnormalities associated with this complex syndrome. With improving knowledge of the human genome, genetic analysis of families with lymphedema continues to offer one of the most promising approaches to identifying genes influencing lymphatic development. PMID- 12543716 TI - Contractility patterns of normal and pathologically changed human lymphatics. AB - Human leg lymphatics contract spontaneously, rhythmically propelling lymph. This intrinsic property regulates the fluid environment in the intercellular space in skin, connective tissue, and perivascular spaces. The pressures generated by lymphatic contractions constitute the main force for lymph flow. This mechanism is of utmost importance during night rest, anesthesia, and immobilization, as well as in those with damaged peripheral motor neurons. All supporting forces are only secondary to those created by spontaneous lymphatic contractions. The intrinsically regulated lymph flow depends on tissue fluid and lymph production rate. The transport capacity of lymphatics adjusts to the fluid load. Under physiological conditions, limb activity and position only slightly change lymph flow. In obstructive lymphedema, high lymph pressures can be observed. They are usually generated by leg muscle contraction, whereas the spontaneous contractility of lymphatics becomes ineffective in lymph transport because of low generated pressures and lymphatic valve insufficiency. The knowledge of lymph flow in normal and lymphedematou limbs will be useful in the derivation of rational treatments for lymphedema. PMID- 12543717 TI - Preclinical models of lymphatic disease: the potential for growth factor and gene therapy. AB - The human disease states that are characterized by functional lymphatic insufficiency currently lack a cure. Molecular approaches may ultimately provide a therapeutic window to reverse the stigmata of both primary and secondary lymphatic insufficiency. To harness the potential therapeutic power of lymphangiogenesis, testing the safety and efficacy of the treatment response will be necessary. This, in turn, necessitates the availability of suitable preclinical animal models of the disease processes in question, along with suitable research tools to permit an assessment of the response to applied therapies. An ideal model would reproducibly and inexpensively replicate the untreated disease of human lymphedema. It would closely simulate the biology, as we understand it, of the human disease, and would replicate both the pathogenesis of the disease, including its natural history and the temporal patterns of its clinical expression. In this way, one might aspire to make valid predictions about the human applicability of therapy by extrapolation from observations in animal models. In addition to the availability of suitable animal models, the required investigative tools must also be available. In the context of lymphangiogenesis, to assess the therapeutic response, one must certainly possess the ability to recognize newly developed lymphatic vasculature. Sophisticated immunohistochemical and imaging techniques make this increasingly feasible. Initial experimental observations indicate that growth factor and gene therapy with VEGF-C holds promise for the treatment of both primary and secondary forms of lymphedema. PMID- 12543719 TI - Placental growth factor (PlGF) and its receptor Flt-1 (VEGFR-1): novel therapeutic targets for angiogenic disorders. AB - Efforts to therapeutically stimulate or inhibit vessel growth have been primarily focused on vascular endothelial growth factor (VEGF) and its receptor VEGFR-2 (Flk-1), while little attention has been devoted to the therapeutic potential for angiogenic disorders of placental growth factor (PlGF), a VEGF family member, and its receptor VEGFR-1 (Flt-1). However, recent developments and insights could shift that focus to P1GF and Flt-1. Indeed, PlGF stimulated angiogenesis and collateral growth in ischemic heart and limb with at least a comparable efficiency to VEGF and did not cause side effects associated with VEGF, such as edema or hypotension. An anti-Flt-1 antibody suppressed neovascularization in tumors and ischemic retina, and angiogenesis and inflammatory joint destruction in arthritis. The anti-Flt-1 antibody also reduced atherosclerotic plaque growth and vulnerability, but the atheroprotective effect was not due to reduced plaque neovascularization. The anti-inflammatory effects of the anti-Flt-1 antibody were attributable to a reduced mobilization of bone marrow-derived myeloid progenitors into the peripheral blood, a reduced mobilization/differentiation (and impaired infiltration) of Flt-1-expressing leukocytes into inflamed tissues, and a defective activation of myeloid cells. Thus, PlGF and Flt-1 constitute potential candidates for therapeutic modulation of angiogenesis and inflammation. PMID- 12543720 TI - Insights into the molecular pathogenesis and targeted treatment of lymphedema. AB - Abnormal function of the lymphatic vessels is associated with a variety of diseases, such as tumor metastasis and lymphedema. The development of strategies for local and controlled induction or inhibition of lymphangiogenesis would thus be of major importance for the treatment of such diseases. Two growth factors, vascular endothelial growth factor C (VEGF-C) and D (VEGF-D), have been found to be important in the proper formation and maintenance of the lymphatic network, through their receptor VEGFR-3. In patients with lymphedema, heterozygous inactivation of VEGFR-3 leads to primary lymphedema due to defective lymphatic drainage in the limbs. We have shown that VEGF-C gene transfer to the skin of mice with lymphedema induces regeneration of the cutaneous lymphatic vessel network. However, as is the case with VEGF, high levels of VEGF-C cause blood vessel growth and leakiness, resulting in tissue edema. Strategies to avoid these side-effects have also been developed. This new field of reseach has important implications for the development of new therapies for human lymphedema. PMID- 12543721 TI - Lymphangiogenesis new mechanisms. AB - A mouse model has been developed to study lymphangiogenesis dissociated from angiogenesis. bFGF implanted in a mouse cornea at a concentration below the threshold to induce angiogenesis potently induces lymphangiogenesis. This model has permitted a study of cellular and molecular mechanisms of lymphangiogenesis. PMID- 12543722 TI - Lymphatic vessel activation in cancer. AB - Metastasis of most cancers occurs primarily through the lymphatic system, and the extent of lymph node involvement is the most important prognostic indicator. While the importance of the lymphatic system as a pathway for metastasis has been well recognized, there is very little information available about the mechanisms by which tumor cells interact with the lymphatics. Recently, production of the lymphangiogenic factor VEGF-C has been detected in tumors, and the significance of VEGF-C-mediated lymphangiogenesis for tumor metastasis has been demonstrated. Increased lymphatic vessel density has been found associated with certain tumors. The mechanisms by which tumor cells gain access to and enter lymphatic vessels are critical issues that need to be addressed in the future. In contrast to the prevailing view that has assigned to the lymphatic system a passive role in the metastatic process, our results indicate the importance of lymphatic vessel activation in tumor dissemination. PMID- 12543723 TI - The pathogenesis of filarial lymphedema: is it the worm or is it the host? AB - Our understanding of the pathogenesis of filarial lymphedema, although evolving, is still limited. Recurrent bacterial infections play a major role in the progression of lymphedema to elephantiasis, but the host and parasite factors that trigger disease development are not known. Field studies in Haiti show that lymphedema and host responses to parasite antigens cluster in families, consistent with the hypothesis that host genes influence lymphedema susceptibility. The recent recognition that filarial parasites harbor the endosymbiotic bacteria, Wolbachia, also raises questions about the potential contribution of the inflammatory response to Wolbachia antigens to lymphedema development. In this review, we discuss potential risk factors for lymphedema and try to integrate these in a model of pathogenesis. PMID- 12543724 TI - The molecular control of adipogenesis, with special reference to lymphatic pathology. AB - Adipogenesis is the process by which mature fat cells are formed from pre adipocytes. Adipogenesis has come under increasing scrutiny not only because the availability of reliable in vitro models makes it an attractive choice for developmental studies, but also because adipocytes are increasingly recognized as major players in a variety of physiological and pathophysiological states, such as obesity and type 2 diabetes. Adipocytes develop from mesenchymal stem cell precursors that are characterized by multipotency. Under the influence of various cues, these cells become committed to the adipocyte lineage. Further hormonal stimulation recruits these pre-adipocytes to accumulate lipid, express fat specific markers, and become sensitive to the metabolic effects of insulin. A complex transcriptional cascade regulates this process, involving several distinct classes of transcription factor. In particular, the role of the nuclear hormone receptor PPARgamma will be discussed, along with bZip family members C/EBPalpha, C/EBPbeta, and C/EBPdelta. The relationship of adipose depots to the lymphatic system will also be discussed. PMID- 12543725 TI - A stepwise model of the development of lymphatic vasculature. AB - Although lymphedema was first described more than a century ago, little progress has been made in understanding the mechanisms that cause it. Investigation of the normal development of the lymphatic system has been hindered by the lack of known lymphatic-specific markers. In 1902, F. Sabin proposed the most widely accepted theory about the origin of the lymphatic vasculature. This model proposed that isolated primitive lymph sacs bud from the endothelium of veins during early development; from these primary lymph sacs, the peripheral lymphatic system spreads by endothelial sprouting into tissues where local capillaries form. In 1999, we identified the homeobox gene Prox1 as the first specific marker of lymphatic endothelial cells. Functional inactivation of Prox1 in mice demonstrated that lymphangiogenesis requires the activity of this gene in a subpopulation of endothelial cells in embryonic veins. Prox1 promotes the development of the lymphatic vasculature by determining the final lymphatic fate of budding venous endothelial cells. On the basis of our findings, we propose a stepwise model of lymphangiogenesis in which lymphatic vasculature development is initiated by the specific expression of Prox1 in a subpopulation of vascular endothelial cells that subsequently adopt a lymphatic vasculature phenotype. PMID- 12543726 TI - De novo lymph node formation in chronic inflammation of the human leg. AB - Organized lymphoid tissue is the first line of antigenic defense. Recruited by antigen located in the non-lymphoid tissues, the infiltrating lymphocytes often organize themselves as follicle-like structures that contain germinal centers, similar to those in secondary lymphoid follicles of lymph nodes. These extranodal tertiary lymphoid follicles are found in various autoimmune diseases. We investigated 153 patients with protracted lymph stasis of the lower limb, caused by lymphatic damage incurred through soft tissue bacterial inflammation or mechanical trauma of soft tissues and bones. In 10% of patients with post inflammatory, and in 25% with post-traumatic lymph stasis, "newly-formed" lymph nodes were detected by means of lymphoscintigraphy. They were located along the large calf and thigh veins. Although scattered nodes are normally detected in these areas, the number and total mass of visualized nodes substantially exceeded those seen in healthy subjects. The calculated surface area of "newly formed" nodes attained 50-70% of the area of ipsilateral inguinal nodes. Histological evaluation of nodal biopsy specimens in three such patients revealed, in one, a lymph node structure without differentiation into cortical and medullary areas, and in a second, a follicle-like structure within a dilated lymph vessel. Lymph clot removed from another dilated vessel contained a lymphocyte/dendritic cell aggregate. The "newly formed" nodes likely originate from primordial lymphoid follicles and/or lymphoid cell aggregates formed in response to chronic stimulation by microbial products and self-antigens from the damaged tissues. Detection of "newly-formed" lymph nodes in the limb is evidence of an ongoing inflammatory process and requires appropriate therapy. PMID- 12543727 TI - Physiologic aspects of lymphatic contractile function: current perspectives. AB - The lymphatic system plays an important role in fluid/macromolecular balance, lipid absorption, and immune functions, and is involved in many different pathologic conditions, like inflammation, spread of cancer cells, and lymphedema. There are several forces that drive lymph centripetally. Extrinsic driving forces, or the passive lymph pump, include lymph formation, arterial pulsations, skeletal muscles contractions, fluctuations of central venous pressure, gastrointestinal peristalsis, and respiration. Intrinsic forces, or the active lymph pump, are the result of coordinated contractions of lymphangions, the morpho-functional units of the lymphatic vessels, which include the valve and portion of the vessel extending to the next valve. The contractions of the lymphangions are initiated by the pacemaker activity of the smooth muscle cells of lymphangion wall. Transmural pressure is an important hydrodynamic factor that modulates pacemaking. Under conditions of low filling, lymphangions might produce negative intraluminal pressures and a suction effect. Because of the complicated hydrodynamic conditions in lymphatic beds, the passive and active lymph pumps sometimes work together to propel lymph centripetally. In other cases (i.e., under conditions of enhanced lymph flow), flow-mediated inhibition of the active lymph pump could serve to decrease lymphatic outflow resistance and save metabolic energy when the driving force of the passive lymph pump is enough to propel lymph. We have recently found that there are profound differences in the pressure and flow sensitivities of lymphatic vessels derived from different tissues, such as the thoracic duct and mesenteric lymphatics. Such results, when considered in light of the controversy surrounding some studies performed in different animals, lead to the idea that the active lymph pumps in humans may have greater regional differences in contractile function than has been seen in animals, because of the upright posture in bipedal humans. This posture creates an additional outflow resistance for lymphatics of the lower part of the body. Thus, despite the ongoing attempts to determine the mechanisms of lymphatic diseases and useful therapies to treat them, there are many disputable or unknown issues regarding the physiology of lymph transport in humans. PMID- 12543729 TI - The role of interstitial stress in lymphatic function and lymphangiogenesis. AB - The management and control of tissue fluid balance depends on the highly regulated orchestration of various interstitial factors. In particular, lymphatic function, lymphatic biology, and development (lymphangiogenesis), and the extracellular matrix all contribute to interstitial fluid balance. In light of the dynamic interdependence of these factors, our lab has been working towards establishing a mechanical-molecular picture of the process of lymphangiogenesis that is, bridging the physiological context of lymphangiogenesis with its molecular regulation by studying the coordination of mechanical forces, ECM development, lymphatic biology, and lymphatic capillary organization and development. Our working hypothesis is that the physiological driving force for lymphangiogenesis is the need for organized interstitial fluid flow. This paper will outline the rationale and background for such an approach and highlight some of the recently published findings of our lab and others that support this concept. PMID- 12543730 TI - Proteomic technologies to study diseases of the lymphatic vascular system. AB - Now that the human genome has been mapped, a new challenge has emerged: deciphering the various products of individual genes. Consequently, new proteomic technologies are being developed to monitor and identify protein function and interactions responsible for the total activities of the cell. The application of these new proteomic technologies to study cellular activities, will lead to a faster sample throughput and increased sensitivity for the detection of individual proteins, thus providing major opportunities for the discovery of new biomarkers for the early detection of protein alterations associated with the progression of the disease state. PMID- 12543733 TI - Suppurative keratitis. PMID- 12543734 TI - Central nervous system mechanisms in Sjogren's syndrome. PMID- 12543735 TI - Ophthalmology in Hong Kong. PMID- 12543736 TI - Corneal blindness in a southern Indian population: need for health promotion strategies. AB - AIM: To assess the distribution and causes of corneal blindness in a population in southern India. METHODS: A total of 11 786 people of all ages from 94 clusters representative of the population of the Indian state of Andhra Pradesh were sampled using a stratified, random, cluster, systematic sampling strategy. These participants underwent a detailed interview and eye examination including measurement of visual acuity with logMAR charts, refraction, slit lamp biomicroscopy, applanation tonometry, gonioscopy, and stereoscopic dilated fundus evaluation. An eye was considered to have corneal blindness if the visual acuity was <20/200 due to a corneal disease. RESULTS: Of those sampled, 10 293 (87.3%) people participated in the study. Corneal blindness in at least one eye was present in 86 participants, an age, sex, and urban-rural distribution adjusted prevalence of 0.66% (95% confidence interval 0.49 to 0.86), which included 0.10% prevalence of corneal blindness in both eyes and 0.56% in one eye. The most frequent causes of corneal blindness in at least one eye included keratitis during childhood (36.7%), trauma (28.6%), and keratitis during adulthood (17.7%). Nearly 95% of all corneal blindness was avoidable. Multivariate analysis showed that the prevalence of corneal blindness was significantly higher with decreasing socioeconomic status and with increasing age. Of the 99 eyes with corneal blindness, 51 (51.5%) had visual acuity of inaccurate projection of light or no perception of light. CONCLUSIONS: There is a significant burden of corneal blindness in this population, the majority of which is avoidable. Eye health promotion strategies are warranted to raise awareness about the causes and prevention of corneal blindness. PMID- 12543738 TI - Community treatment with azithromycin for trachoma is not associated with antibiotic resistance in Streptococcus pneumoniae at 1 year. AB - AIMS: To determine if macrolide resistant Streptococcus pneumoniae will be a major concern in areas that receive annual mass azithromycin distributions for trachoma. METHODS: A cross sectional survey was conducted of nasopharyngeal S pneumoniae isolates for susceptibility to azithromycin 1 year after administering a single dose of azithromycin to treat trachoma in a village in Nepal. RESULTS: S pneumoniae was isolated from 50 (86%) of 57 nasopharyngeal cultures and no resistance to azithromycin was detected. CONCLUSION: The authors were unable to demonstrate that mass azithromycin therapy for trachoma produced macrolide resistant S pneumoniae that persists until the next scheduled annual treatment. PMID- 12543737 TI - Follicular conjunctivitis caused by Chlamydia trachomatis in an infant Saharan population: molecular and clinical diagnosis. AB - AIMS: To ascertain the prevalence of acute trachoma as a supposed endemic disease among infants in the Saharan refugee camps of Tindouf (Algeria) and to evaluate the efficacy of treatment with a single dose of azithromycin. METHODS: 527 children aged between 3 and 17 were evaluated in the camp schools in October 2001. All the children were clinically and microbiologically examined, including slit lamp checks of anterior segment and two conjunctival swabs, one for the detection of membrane lipopolysaccharide by quick immunoassay test Clearview Chlamydia MF and the other for the detection of specific DNA by chlamydia plasmid specific polymerase chain reaction (PCR) assay. After examination, a single dose of azithromycin was administered to each child. One month later samples from positive children and 12 random negative children by the first PCR were taken to perform a new PCR assay. RESULTS: Chlamydia trachomatis was suspected in 2.47% of the children, papillary hypertrophy being the most frequently seen clinical sign. Scarring changes secondary to trachoma were detected in 11.7% of the children. Only four cases (0.8%) were positive to the immunoassay test and 12 cases (2.27%) were positive by PCR. After treatment a second PCR was performed on positive children and they were negative of chlamydia DNA amplification. However, one child who was negative and received the treatment was positive in the second PCR assay. CONCLUSION: Acute trachoma prevalence is much lower than expected among infants in this Saharan population. The possible reasons could be the recent improvements in hygiene and health care. Cicatricial trachoma is more frequent. The Clearview immunoassay test has very low sensitivity in the detection of this disease. A single dose of azithromycin is sufficient to treat chlamydial conjunctivitis. However, a programme of improvement in hygiene and health care is necessary to prevent re-infection. PMID- 12543739 TI - The phenotype of normal tension glaucoma patients with and without OPA1 polymorphisms. AB - AIM: Polymorphisms in OPA1, the gene responsible for autosomal dominant optic atrophy, were recently found to be strongly associated with normal tension glaucoma (NTG). The aim of this study was to determine whether OPA1 polymorphisms affect the phenotype of NTG patients. METHODS: A retrospective analysis was performed of 108 well characterised NTG patients who had been genotyped for OPA1 variations, and who had previously undergone automated perimetry and Heidelberg retina tomography (HRT). 25 NTG patients had the at-risk OPA1 genotype (IVS 8 +4 C/T; +32 T/C) and 83 NTG patients did not. Differences between groups were sought in a wide range of structural, psychophysical, and demographic factors. These included sex, age at diagnosis, family history of glaucoma, history of ischaemic risk factors and vasospasm, laterality of glaucoma, presenting and highest diurnal intraocular pressure (IOP), initial cup-disc (CD) ratio, baseline visual field global indices, and optic disc parameters as measured by HRT. For a subgroup of patients with at least 5 years of follow up and 10 visual field tests, pointwise linear regression analysis (PROGRESSOR for Windows software) was applied to the visual field series. RESULTS: There was no significant difference in the two groups with respect to sex, age at diagnosis, family history of glaucoma, history of ischaemic risk factors and vasospasm, or laterality of glaucoma. The comparison of IOP, CD ratio and visual field global indices, MD and CPSD in the two groups showed no significant difference. There were no differences in the mean values for any of the HRT parameters analysed. For the subgroup of patients with at least 5 years of follow up, there was also no significant difference in the number of patients with progressing locations, the mean number of progressing locations per subject, the mean slope of the progressing locations or the mean slope for whole visual field. CONCLUSIONS: The absence of phenotypic differences in normal tension glaucoma patients with and without the OPA1 polymorphisms IVS 8 +4 C/T; +32 T/C suggest that these OPA1 polymorphisms do not underlie any major phenotypic diversity in these patients. PMID- 12543740 TI - Effect of a patient training video on visual field test reliability. AB - AIMS: To evaluate the effect of a visual field test educational video on the reliability of the first automated visual field test of new patients. METHODS: A prospective, randomised, controlled trial of an educational video on visual field test reliability of patients referred to the hospital eye service for suspected glaucoma was undertaken. Patients were randomised to either watch an educational video or a control group with no video. The video group was shown a 4.5 minute audiovisual presentation to familiarize them with the various aspects of visual field examination with particular emphasis on sources of unreliability. Reliability was determined using standard criteria of fixation loss rate less than 20%, false positive responses less than 33%, and false negative responses less than 33%. RESULTS: 244 patients were recruited; 112 in the video group and 132 in the control group with no significant between group difference in age, sex, and density of field defects. A significant improvement in reliability (p=0.015) was observed in the group exposed to the video with 85 (75.9%) patients having reliable results compared to 81 (61.4%) in the control group. The difference was not significant for the right (first tested) eye with 93 (83.0%) of the visual fields reliable in the video group compared to 106 (80.0%) in the control group (p = 0.583), but was significant for the left (second tested) eye with 97 (86.6 %) of the video group reliable versus 97 (73.5%) of the control group (p = 0.011). CONCLUSIONS: The use of a brief, audiovisual patient information guide on taking the visual field test produced an improvement in patient reliability for individuals tested for the first time. In this trial the use of the video had most of its impact by reducing the number of unreliable fields from the second tested eye. PMID- 12543742 TI - Efficacy of lignocaine 2% gel in chalazion surgery. AB - BACKGROUND/AIMS: To determine whether topical 2% lignocaine (lidocaine) gel is an effective anaesthetic agent for chalazion surgery. METHODS: In a randomised controlled clinical trial, 57 subjects aged 12 years or over requiring incision and curettage for chalazion were recruited over an 8 month period. Patients were randomised into two groups. One group received 1.5 ml of lignocaine 2% injection and the other 1.5 ml of lignocaine 2% gel topically. Standard incision and curettage was then performed. The primary outcome of interest was the total pain experienced during the entire procedure including anaesthetic administration as well as incision and curettage. The pain from the local anaesthetic administration and during incision and curettage was assessed independently using a visual analogue scale (0-100). The sum of these two scores would be the total pain score out of 200. "Fear of injection" score (0-100) was also assessed. RESULTS: There was a statistically significant difference in the mean total pain scores between the injection and the gel groups (95.6 v 57.0) (p <0.001) (alpha = 0.05) (1 - beta = 0.9394). There was a statistically significant difference in the mean scores on "pain of anaesthetic administration" (47.0 v 5.5) (p <0.000). There was no statistically significant differences in the mean scores on "fear of injection" (43.9 v 47.7) (p = 0.668) and "pain during incision and curettage" (48.28 v 51.4) (p=0.679). CONCLUSIONS: Lignocaine 2% gel is effective in chalazion surgery especially in lowering the pain caused by anaesthetic administration. PMID- 12543743 TI - Posterior corneal topographic changes after partial flap during laser in situ keratomileusis. AB - AIM: To study the posterior corneal topographic changes in eyes with partial flaps during laser assisted in situ keratomileusis (LASIK). METHODS: Case records of 16 patients, who had partial flap in one eye during LASIK (group 1) and uncomplicated surgery in the other eye (group 2), were studied. Following occurrence of partial flap intraoperatively, laser ablation was abandoned in all the eyes. A 160/180 micro m flap was attempted during the initial procedure using the Hansatome microkeratome (Bausch & Lomb Surgicals, Munich, Germany). LASIK surgery in all cases was performed using a 180 micro m plate, at the mean interval of 4.16 (SD 1.5) months following the initial procedure. None of the eyes had intraoperative complication during LASIK. Relative posterior corneal surface elevation above the best fit sphere (BFS) before the initial procedure, before, and after LASIK were compared using the Orbscan slit scanning corneal topography/pachymetry system. RESULTS: Posterior corneal elevation was comparable in the two groups, both preoperatively (group 1; 16.4 (4.8) micro m, group 2; 16.1 (4.8) micro m) and after final surgery (group 1; 57.2 (15.6) micro m, group 2; 54.3 (13.1) micro m). In group 1 after occurrence of partial flap, the posterior corneal elevation was 16.9 (4.4) micro m, and this increase was not significant statistically (p=0.4). On multiple linear regression analysis, residual bed thickness (p<0.001) was independently the significant determinant of final posterior corneal elevation in both groups. CONCLUSION: The inadvertent occurrence of partial flap during LASIK procedure does not contribute to the increase in posterior corneal elevation. PMID- 12543744 TI - Comparison of 5% povidone-iodine solution against 1% povidone-iodine solution in preoperative cataract surgery antisepsis: a prospective randomised double blind study. AB - BACKGROUND/AIM: Povidone-iodine (PI, Betadine) is routinely used as a preoperative topical antiseptic in cataract surgery as it has been shown to reduce the incidence of postoperative endophthalmitis. However, the concentration used clinically is variable. In vitro studies have shown that PI is paradoxically more effective at lower concentration. This study was undertaken to determine if this effect was reproducible in vivo. METHODS: A prospective randomised double blind study was carried out in the ophthalmic theatre in a district general hospital. 105 patients attending for routine cataract surgery were randomly allocated to have their conjunctival fornices irrigated preoperatively with either PI 1% (group A) or PI 5% (group B). Conjunctival swabs were taken, in identical fashion, both before and 1 minute after irrigation. The number and species of bacterial colonies cultured from each swab was counted. The difference in the median number of bacterial colonies from pre-irrigation to post-irrigation cultures was then compared between the groups. RESULTS: Bacterial cultures were gained from 100 patients (33 male, 67 female, mean age 74 years, range 30-95 years). Group B (5% PI) showed a decrease in median colony forming units (CFU) pre-irrigation from 100 to 40 CFU post-irrigation (a drop of 60%). This was greater than in group A (1% PI) where the reduction was 120 CFU pre-irrigation to 100 CFU post-irrigation (a drop of 16.7%) (Mann-Whitney test, p<0.05). At higher initial bacterial loads (CFU pre-irrigation >1000), the difference in median between the two groups became larger as the number of pre-irrigation bacteria increased. In group B pre-irrigation CFU reduced from 3340 to 110 post-irrigation (a drop of 96.7%) compared with group A: 5000 CFU pre-irrigation to 3000 post irrigation (a drop of 40%) (Mann-Whitney test, p=0.0014). CONCLUSION: Despite in vitro evidence of higher bactericidal efficacy of PI at more dilute concentrations, 5% PI is more effective than 1% PI in decreasing the human conjunctival bacterial flora in vivo, particularly in the presence of heavier initial bacterial load. PMID- 12543745 TI - Five year incidence of cataract surgery: the Blue Mountains Eye Study. AB - AIMS: To assess the 5 year incidence of cataract surgery in an older population based prospective cohort. METHODS: 5 Year prospective follow up of the population based Blue Mountains Eye Study (BMES) performed in 1992. The follow up study examined 2335 survivors (75.1%) of the 3654 baseline participants. Baseline and 5 year slit lamp and retroillumination lens photographs were graded for presence of cortical, nuclear, or posterior subcapsular cataract using the Wisconsin cataract grading method and cataract surgery was documented from the history and the clinical examination. RESULTS: An overall cataract surgery rate of 5.7% in first or both eyes was documented. The incidence was 0.3% in people aged 49-54 years at baseline, 1.7% for ages 55-64 years, 7.9% for ages 65 to 74 years, and 17.4% in people aged 75 years or older. The rate of surgery in first or both eyes was 6.0% in women and 5.2% in men, age adjusted p = 0.66. Bilateral cataract surgery was performed during follow up on 2.7% of participants, while 43.1% of unilateral phakic cases had second eye surgery. Presence of any posterior subcapsular (PSC) cataract, either alone or in combination with other cataract types, was the most likely type of cataract at baseline to be associated with incident cataract surgery. Baseline age was the most important non-ocular variable predicting incident cataract surgery. CONCLUSIONS: This study has documented age specific rates for 5 year incident cataract surgery in an older community. The finding of relatively similar incidence rates and ocular predictors of cataract surgery to those reported by the Beaver Dam Eye Study, Wisconsin, United States, is of interest, given previous documented similarities between these two populations. PMID- 12543746 TI - Verteporfin photodynamic therapy in highly myopic subfoveal choroidal neovascularisation. AB - AIMS: To analyse the visual and angiographic results of photodynamic therapy (PDT) with verteporfin in highly myopic patients with subfoveal choroidal neovascularisation (CNV). METHODS: PDT was performed on highly myopic patients with subfoveal CNV. The patient population was divided into two groups according to age at onset of CNV (group 1 aged < or = 55 and group 2 aged >55 years old). Best corrected visual acuity (BCVA) and angiographic findings were considered. RESULTS: At the end of follow up group 2 had worse BCVA than group 1. 76% of patients in both groups showed a complete closure of CNV at the end of follow up. CONCLUSION: Visual prognosis of myopic CNV treated by PDT is influenced by age at onset. PMID- 12543747 TI - Angiographic features after photodynamic therapy for choroidal neovascularisation in age related macular degeneration and pathological myopia. AB - AIM: To describe the angiographic features after photodynamic therapy (PDT) with verteporfin in choroidal neovascularisation (CNV) associated both with age related macular degeneration (AMD) and pathological myopia (PM). METHODS: 36 patients affected by subfoveal CNV in AMD and 25 patients with subfoveal CNV in PM underwent an ophthalmological examination including fluorescein angiography (FA) and indocyanine green angiography (ICGA) using the IMAGEnet System. Post-PDT examinations were performed 7, 30, and 90 days later. RESULTS: The typical angiographic aspect after PDT for AMD related CNV was a round hypofluorescence visible both on FA and on ICGA, which included both CNV and the surrounding tissues and corresponded to the area exposed to laser light. In PM the CNV appeared hypofluorescent during the early phases and gradually became hyperfluorescent during the late phases on FA, whereas on ICGA it was detectable in its whole extension as a hyperfluorescent lesion since the early phases. Differently from AMD, there was no round hypofluorescence surrounding the CNV on FA or on ICGA. Moreover, five patients in the AMD group showed hot spots on ICGA, which spontaneously disappeared during the follow up. Classic and occult components of the AMD related CNV revealed a different angiographic response to PDT, showing with the latter only a partial closure 1 week after PDT followed by a complete reopening at the first month in 100% of cases. CONCLUSION: The post PDT hypofluorescence typical of AMD related CNV, especially visible on FA, might be secondary to a combination of choriocapillary occlusion and masking effect due to swelling of retinal pigment epithelium cells. Hot spots in the AMD affected patients could be interpreted as the expression of a non-thermal choroidal vasculitis secondary to PDT. PMID- 12543748 TI - Blood flow per unit retinal nerve fibre tissue volume is lower in the human inferior retina. AB - AIM: To determine if perfusion per unit tissue volume of retinal nerve fibre layer and optic nerve head in the inferior sector is lower than in the superior sector. METHODS: Heidelberg retinal tomogram (HRT) for topographic measurement of optic nerve head and retinal nerve fibre layer and Heidelberg retinal flowmeter (HRF) for retinal blood flow were performed on 19 normal healthy subjects. Measurements from the superior and inferior sectors were compared. The perfusion/nerve fibre ratio (PNR); the blood flow per unit retinal nerve fibre tissue volume, was calculated in each sector with a formula; HRF flow measurements divided by HRT measurements. RESULTS: Retinal nerve fibre layer thickness in the inferior retina was significantly higher than in the superior retina (p<0.05). There were, however, no differences in retinal blood flow between the superior and inferior retinal sectors. The PNR in the inferior sector were significantly lower than in the superior sector (p=0.047 for HRF mean flow/rim volume and p = 0.0282 for HRF 75th percentile flow/rim volume). CONCLUSIONS: The inferior sector of retinal nerve fibre layer and optic nerve head may have lower blood flow per unit nerve tissue volume compared to the superior sector. This result suggests that the inferior sector is more vulnerable to elevated intraocular pressure (IOP) and ischaemic insults in glaucomatous optic neuropathy. PMID- 12543749 TI - Optic disc morphology in south India: the Vellore Eye Study. AB - AIM: To evaluate the morphology of the optic nerve head in an unselected population group in south India. METHODS: The study included 70 subjects forming a population based sample, selected in a random manner. Mean age was 47.5 (SD 8.7) years, mean refractive error measured -0.07 (1.11) dioptres (range -4.50 to +2.50 dioptres). Optic disc slides were morphometrically analysed. RESULTS: Mean optic disc area measured 2.58 (0.65) mm(2). It was statistically independent of age and refractive error. Optic disc shape was slightly vertically oval. Mean neuroretinal rim area was 1.60 (0.37) mm(2). It was significantly and positively correlated with optic disc size and optic cup size. It was independent of age, sex, refractive error, and axial length. In all subjects included in the study, the rim was smallest in the temporal horizontal optic disc sector. Mean horizontal cup/disc diameter ratio (0.66 (0.07)) was significantly (p<0.001) higher than the mean vertical cup/disc diameter ratio (0.56 (0.08)). Both ratios were highly significantly (p <0.001) and positively correlated with optic disc size. The alpha zone of parapapillary atrophy (0.84 (0.29) mm(2)), and beta zone (0.13 (0.38) mm(2)), respectively, occurred in 69 (98.6%) subjects and in eight (11.4%) subjects, respectively. They were significantly larger in the temporal horizontal sector. The alpha zone was significantly (p<0.001) larger and occurred significantly more often than beta zone. Retinal arterioles and venules were wider, and in spatial correlation, the visibility of the retinal nerve fibre layer was significantly better, in the temporal inferior disc arcade and the temporal superior arcade than in the nasal superior arcade and the nasal inferior vessel arcade. Except for the absolute size measurements these optic nerve head parameters did not differ markedly (p >0.05) from the values found in white people. CONCLUSIONS: South Indians and white people do not show marked differences in the morphology of the optic nerve head as measured by morphometric optic disc parameters, with the possible exception of the absolute optic disc dimensions. PMID- 12543751 TI - A clinical and molecular genetic study of a rare dominantly inherited syndrome (MRCS) comprising of microcornea, rod-cone dystrophy, cataract, and posterior staphyloma. AB - AIM: To phenotype and genetically map the disease locus in a family presenting with autosomal dominant microcornea, rod-cone dystrophy, cataract, and posterior staphyloma. METHODS: Six affected and three unaffected members of the pedigree were examined. All individuals provided a history and underwent a full clinical examination with A-scan and B-scan ultrasonography and electrophysiological testing where appropriate. PCR based microsatellite marker genotyping using a positional candidate gene approach was then performed on DNA samples extracted from venous blood provided by each subject. RESULTS: The disorder is inherited as an autosomal dominant trait with variable expressivity and has a complex phenotype. Affected individuals had bilateral microcornea, pulverulent-like lens opacities, a rod-cone dystrophy and posterior staphyloma (MRCS). Using a positional candidate gene approach, the authors have evidence suggestive of linkage of this disorder to a region on 11q13 within the nanophthalmos 1 (NNO1) genetic interval. The small family size militates against achieving a LOD score of 3, but the haplotype data and the position of the putative MRCS locus within a known nanophthalmos locus are suggestive of linkage. A candidate gene within this region (ROM1) was screened and no mutations were found in affected members of the family. CONCLUSION: This rare developmental disorder has some phenotypic similarities to nanophthalmos and possibly maps to a locus within the genetic interval encompassing the NNO1 locus. Screening of candidate genes within this region continues. PMID- 12543752 TI - Intercellular adhesion molecule-1 expression on human corneal epithelial outgrowth from limbal explant in culture. AB - AIM: To investigate the relation between intercellular adhesion molecule (ICAM)-1 expression and cellular dynamics occurring concomitantly with epithelial cell movement. METHODS: Outgrowing epithelial sheets of human corneal epithelial (HCE) cells from cultured limbal explants were examined by immunoperoxidase staining with anti-ICAM-1 monoclonal antibody. An adhesion assay was performed using the epithelial sheets of HCE cells and an Epstein-Barr virus (EVB) infected B cell lymphoma cell line (EVB(+)BJAB) expressing CD11a/CD18, a counter-receptor of ICAM 1. Also, the effect of calphostin C, a specific protein kinase C (PKC) inhibitor, on ICAM-1 expression on the outgrowing epithelial sheets of HCE cells was examined. RESULTS: Strong positive staining for ICAM-1 was found predominantly on HCE cells in the marginal segment of the epithelial sheet, particularly on the cells at the leading edge. EBV(+)BJAB cells adhering to the HCE cells corresponded well to the area of ICAM-1 staining. Treatment of outgrowing epithelial sheets with calphostin C markedly decreased the ICAM-1 expression on the HCE cells. CONCLUSION: ICAM-1 is actively expressed on HCE cells in the marginal segment of the outgrowing epithelial sheets where there is active movement mediated through a PKC dependent mechanism, suggesting the role of ICAM 1 in epithelial cell motility such as the spreading and migration of cells. PMID- 12543753 TI - Expression and distribution of matrix metalloproteinases and their inhibitors in the human iris and ciliary body. AB - AIM: To determine the expression and distribution of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) in the normal human iris and ciliary body. METHODS: Seven postmortem human eyes were fixed with formalin. The iris and ciliary body were dissected out and embedded in paraffin. The expression of MMPs 1, 2, 3, and 9, and TIMPs 1-4 in the iris and ciliary body was determined by a novel immunofluorescence technique and the results graded by masked observers. RESULTS: Positive staining for MMPs and TIMPs was observed in all regions of the anterior uvea, and was more intense in the ciliary body than in the iris. Most MMPs and TIMPs showed similar patterns in their distribution. In the ciliary body, staining was strongest in the epithelium, and was localised to the epithelial cell cytoplasm, except for TIMP-3 which was strongly expressed in the basement membranes. In the iris, staining was most noticeable in the anterior border and anterior epithelial layer. Blood vessels in the stroma of the iris and ciliary body also stained moderately for MMPs and TIMPs. CONCLUSION: Both MMPs and TIMPs are widely expressed in the anterior uvea, with a positive correlation between their expressions. Their differential localisation in the ciliary body suggests they may have a role in maintaining homeostasis in the uveal tract. PMID- 12543754 TI - Distribution of the collagen IV isoforms in human Bruch's membrane. AB - AIMS: To determine the distribution of the alpha1 to alpha6 chains of type IV collagen in Bruch's membrane of the human posterior pole. METHODS: Cryosections (10 micro m) from 18 human eyes (20 months to 83 years old) were acid treated, blocked with 10% normal goat serum, incubated for 1 hour with monoclonal antibodies against type IV collagen isoform specific peptides at 1:75 dilution, and visualised with an ABC staining kit. RESULTS: In Bruch's membrane, the alpha1(IV) and alpha2(IV) chains were identified in retinal pigment epithelial (10/18 = 55%) and choriocapillaris basement membranes (18/18 = 100%); the alpha3(IV), alpha4(IV), and alpha5(IV) chains were also found in the retinal pigment epithelial basement membrane (13/18 = 72%). In the choroid, the alpha1(IV) and alpha2(IV) chains were detected in the blood vessels (18/18=100%). The alpha6(IV) chain was not identified in any sections. CONCLUSION: The heterogeneous distribution of alpha1-2(IV) and alpha3-5(IV) in Bruch's membrane could give insights into the function of this structure in health, ageing, and diseases such as age related macular degeneration. PMID- 12543755 TI - Trypan blue staining of internal limiting membrane and epiretinal membrane during vitrectomy: visual results and histopathological findings. AB - AIMS: To report on the use of trypan blue (TB) 0.06% for staining the internal limiting membrane (ILM) and epiretinal membrane (ERM) during vitrectomy and report on their histology. METHOD: 14 consecutive patients with idiopathic macular hole or macular pucker (seven patients each) were prospectively recruited for ILM or ERM peel respectively. After pars plana vitrectomy and induction of posterior vitreous detachment, 0.5 ml TB 0.06% in phosphate buffered saline (VisonBlue) was injected over the posterior pole in an air filled eye and left for 2 minutes. The stained tissue was peeled with intraocular forceps. Specimens were evaluated using histochemical and immunohistochemical methods. RESULTS: The average follow up was 4.4 months. Internal limiting membranes and epiretinal membranes were stained satisfactorily in all cases and removed successfully. Eight patients (57%) had improvement of 2 or more Snellen lines. All seven macular holes closed. In the ERM cases, no residual membranes were observed clinically, at the latest follow up. No complications relating to the use of the dye were encountered intraoperatively or postoperatively. Of the 14 procedures, nine (four macular hole and five macular pucker) yielded sufficient tissue for histopathological evaluation. Histological and immunohistological assessment revealed that the morphology of these specimens was similar to that observed in macular hole ILM and macular pucker ERM removed without the aid of dye. CONCLUSION: TB staining facilitated the identification and delineation of ILM and ERM removal during the surgical management of macular holes and macular pucker. The visual outcome of this series and the specimens removed suggest they are no different from those without TB staining. Its use in posterior segment appears to be safe but further studies are required to investigate its long term safety. PMID- 12543756 TI - Repair of the lax medial canthal tendon. PMID- 12543759 TI - Merkel cell carcinoma of the eyelid in association with chronic lymphocytic leukaemia. PMID- 12543760 TI - Steatocystoma simplex of the caruncle. PMID- 12543761 TI - Preliminary results with posterior lamellar keratoplasty for endothelial failure. PMID- 12543762 TI - Persistent accommodative spasm after severe head trauma. PMID- 12543757 TI - In vivo confocal microscopy of the human cornea. AB - AIMS: To describe the optics of in vivo confocal microscopy, its advantages over previous methods, and to summarise the literature that arose from its use for the observation of the human cornea. A critical review of the clinical usefulness of this new technology for the corneal examination is undertaken. METHODS: Confocal microscopes obtain increased resolution by limiting the illumination and observation systems to a single point. Rapid scanning is used to reconstruct a full field of view and allows for "real time" viewing. RESULTS: Coronal sections of the in situ epithelium, Bowman's membrane, stroma, and endothelium can be visualised at a resolution of 1-2 micro m. A backscattered light intensity curve allows objective measurements of sublayer thickness and corneal haze to be taken. In vivo confocal microscopy is therefore particularly useful in the areas of infective keratitis, corneal dystrophies, refractive surgery, and contact lens wear, where it aids in differential diagnosis and detection of subtle short and long term changes. Real time endothelial cell assessment can also be performed. CONCLUSION: Because of their ability to visualise living tissue at cellular levels, confocal microscopes have proved useful additions to the current clinical tools. PMID- 12543763 TI - Extramedullary plasmacytoma of the eyelid. PMID- 12543764 TI - Idiopathic sclerochoroidal calcification. PMID- 12543765 TI - Stereotactic irradiation of biopsy proved optic nerve sheath meningioma. PMID- 12543766 TI - Severe interferon associated retinopathy. PMID- 12543767 TI - Mass lesions of the posterior segment associated with Bartonella henselae. PMID- 12543768 TI - Corneal opacification following keratoplasty in the rat model. PMID- 12543769 TI - Optic neuritis with marked distension of the optic nerve sheath due to local fluid congestion. PMID- 12543770 TI - Surgery for glaucoma in the 21st century. PMID- 12543772 TI - Micro-magnetic resonance lymphangiography in mice using a novel dendrimer-based magnetic resonance imaging contrast agent. AB - Major advances in cancer biology and immunology have been gained using mouse models. However, very few methods are currently available to visualize the deep lymphatic system. A new micro-magnetic resonance lymphangiography (MRL) method in mice, which uses dendrimer-based magnetic resonance imaging contrast agents, was developed. Micro-MRL imaging clearly visualized most of the mouse lymphatic system, including both lymphatics and lymph nodes. This method could detect and distinguish among dilation of lymphatic vessels in a lymphangitis model, proliferative or neoplastic lymph node swellings in a lymphoproliferative model, and inflammatory lymph node swellings in an infection/inflammation model. Changes in the lymphoid system of transgenic mice overexpressing interleukin-15 could be visualized. Abnormal enlarged lymph nodes identified by micro-MRL were selectively removed and analyzed to demonstrate their cell type, receptor expression, and clonality in individual mice. We conclude that the enhanced resolution of this noninvasive micro-MRL can detect and classify lymphatic and lymph node abnormalities in mice, which should have wide applicability to the study of immunology and cancer in both experimental animals and clinical medicine. PMID- 12543773 TI - Overexpression of a dominant negative type II bone morphogenetic protein receptor inhibits the growth of human breast cancer cells. AB - Bone morphogenetic proteins (BMPs) exert cell type-specific effects on cell proliferation. To clarify the role of the BMP pathway in human breast cancer cells, we used a dominant negative strategy with a truncated human type II BMP receptor (DN-BMPRII; amino acid 1-172) fused to the NH2 terminus of enhanced green fluorescent protein. Transient overexpression of DN-BMPRII interfered with BMP-2-induced Smad1 transcriptional activity and caused cells to accumulate in G1. Stable cell lines that constitutively overexpressed DN-BMPRII were resistant to BMP-2-induced Smad1 phosphorylation and proliferated much more slowly than control stable cell lines. These results suggest that BMPs interacting with type II BMP receptors contribute to the proliferation and/or survival of human breast cancer cells. PMID- 12543774 TI - Increased nuclear phosphatase and tensin homologue deleted on chromosome 10 is associated with G0-G1 in MCF-7 cells. AB - Phosphatase and tensin homologue deleted on chromosome 10 (PTEN) is a tumor suppressor that causes cell cycle arrest. Lack of a nuclear locator sequence and a function in the cytosolic phosphatidylinositol 3'-kinase/Akt pathway diverted its study from the nucleus. However, immunohistochemistry revealed PTEN in the nucleus of normal cells and decreased nuclear PTEN in neoplastic tissues. Using protein expression analysis and fluoroscopic localization of green fluorescence protein-tagged PTEN, we examined nuclear PTEN in MCF-7 cells. We demonstrate that PTEN enters the nucleus and that nuclear PTEN varies throughout the cell cycle. Higher nuclear PTEN levels were associated with G0-G1 phase, and lower nuclear PTEN levels were associated with S phase. We postulate that nuclear PTEN activity might directly regulate the cell cycle. PMID- 12543775 TI - Recombinant osteoprotegerin decreases tumor burden and increases survival in a murine model of multiple myeloma. AB - The aim of the present study was to determine whether modifying the local bone environment with osteoprotegerin (OPG), the soluble decoy receptor for receptor activator of nuclear factor-kappaB (RANK) ligand, could affect tumor burden and survival in the 5T33MM murine model of multiple myeloma. Treatment of mice, injected with 5T33MM cells, with recombinant OPG (Fc-OPG) caused a significant decrease in serum paraprotein and tumor burden and a significant increase in time to morbidity. This was associated with a decrease in osteoclast number in vivo but had no effect on apoptosis and proliferation of 5T33MM cells in vitro. These data indicate that targeting the bone microenvironment by inhibiting the interaction between RANK ligand and RANK with Fc-OPG not only inhibits the development of myeloma bone disease but also decreases tumor growth and increases survival. PMID- 12543776 TI - Apoptosis caused by chemotherapeutic inhibition of nuclear factor-kappaB activation. AB - Both the protein kinase C (alpha/beta) inhibitor Go6976 and expression of dominant-negative nuclear factor (NF)-kappaB inhibitor kinase mutants: (a) blocked the growth and caused regression of a mammary tumor insyngeneic mice; (b) inhibited epidermal growth factor (EGF)-induced activation, nuclear translocation, and DNA-binding activity of NF-kappaB; and (c) caused apoptosis of EGF-stimulated cultured mammary tumor cells. cDNA microarray analysis revealed that these treatments reversed the expression changes of a subset of genes altered by EGF treatment. These included: up-regulation of proapoptotic genes of the tumor necrosis factor (TNF) pathway, death-associated protein (DAP) kinase, p53, and p21/Waf1; and down-regulation of inhibitors of apoptosis: inhibitor of apoptosis(IAP)-1 and X-IAP, TNF receptor-associated factor (TRAF)-2, and factors OX40 and 4-1BB. These results and our previous studies suggest the practicality of a target-directed chemotherapy for EGF-responsive breast cancers, by blocking NF-kappaB activation and thereby reinstating apoptosis. PMID- 12543777 TI - Insulin-like growth factor II receptor gene-167 genotype increases the risk of oral squamous cell carcinoma in humans. AB - Our purpose was to evaluate inherited short tandem repeat polymorphisms of the insulin-like growth factor II receptor gene (IGF2R) in oral cancer risk. The 197 individuals that consented to participate in a hospital-based, case-control study were interviewed with a structured questionnaire and provided blood and saliva. DNA was extracted for genotyping using a PCR-based method. Odds ratios were calculated using multivariate logistic regression. Subjects carrying the heterozygous 167-bp IGF2R genotype had a 2.7-fold higher risk of oral cancer compared with subjects with other genotypes (odds ratio = 2.7, 95% confidence interval: 1.16-6.48), controlling for major confounders. Our results suggest that genetic variation of IGF2R may influence significantly the risk of oral cancer. PMID- 12543778 TI - Bcr and Abl interaction: oncogenic activation of c-Abl by sequestering Bcr. AB - c-Abl tyrosine kinase is under rigorous control because of an unknown cellular inhibitor that maintains c-Abl in a relatively inactive state. Because SH2 domains are positive regulators of the nonreceptor tyrosine kinases, we tested whether this putative inhibitor would bind to an Abl SH2 protein construct and thus activate the c-Abl tyrosine kinase. Expression of a Mr 10,000 Abl SH2 protein in COS-1 and Rat-1 cells activated the tyrosine kinase activity of p145 ABL and induced both morphological transformation and foci formation in Rat-1 cells. Importantly, the R to L mutant of the FLVRES sequence of the Abl SH2 protein also activated the c-Abl tyrosine kinase and induced oncogenic transformation. Addition of the Abl kinase inhibitor STI-571 to ABL SH2 transformed Rat-1 cells inhibited tyrosine phosphorylation of p145 ABL. Overexpression of Bcr has been shown to inhibit the Bcr-Abl oncoprotein, and the endogenous Bcr protein forms a complex with c-Abl in hematopoietic cells and insect cells. Therefore, we determined whether Bcr is the putative c-Abl inhibitor that interacts with the Mr 10,000 Abl SH2 protein. Bcr expression in Rat-1 cells transformed by the Mr 10,000 Abl SH2 protein reduced the activated c Abl tyrosine kinase activity to near normal levels and reversed the oncogenic effects (morphology changes and foci formation) seen in the Abl SH2-treated cells. We additionally demonstrated that Bcr and the Mr 10,000 Abl SH2 protein are present in a complex. We conclude from these studies that Bcr is a major tyrosine kinase inhibitor of cytoplasmic c-Abl and that procedures that sequester Bcr will release the c-Abl protein from the Bcr/c-Abl complex, which leads to c Abl oncogenic activation. PMID- 12543779 TI - BRD4-NUT fusion oncogene: a novel mechanism in aggressive carcinoma. AB - The poorly differentiated carcinoma with t(15;19)(q13, p13.1) is characterized by its highly aggressive, invariably lethal clinical course. The chromosome 19 translocation breakpoint targets the BRD4 double bromodomain-containing gene, which functions in regulation of cell cycle progression. Herein we demonstrate that BRD4 is fused with nearly the entire transcript of the novel 15q13 gene, NUT (nuclear protein in testis), forming a 6.4-kb fusion oncogene, BRD4-NUT. NUT, like BRD4, is predicted to encode a nuclear protein but, unlike the ubiquitous BRD4 transcript, is expressed only in testis. These findings establish a model to elucidate the oncogenic consequences of unscheduled NUT expression and altered BRD4 function. Very few fusion oncogenes have been identified in epithelial tumors, and BRD4-NUT is the first fusion oncogene mechanism identified in a highly lethal form of carcinoma. PMID- 12543780 TI - Prolonged treatment with angiostatin reduces metastatic burden during radiation therapy. AB - Ionizing radiation (IR) and concomitant angiostatin (AS) produce greater than additive local antitumor effects. We examined whether prolonged AS treatment added to IR reduces proliferation of lung metastases from LLC primary tumors. Flank tumors were treated with 40 Gy with or without AS (25 mg/kg/day). IR plus a 14-day course of AS improved local tumor control and blocked the increase in lung weights observed in the group receiving IR plus a 2-day course of AS group. Animals treated with prolonged AS exhibited no increase in lung weight and no macrometastases. These findings suggest that long-term treatment with antiangiogenic compounds may be effective in preventing metastases from IR treated tumors as well as increasing the local antitumor effects of radiotherapy. PMID- 12543781 TI - Inhibition of glutathione synthesis reverses Bcl-2-mediated cisplatin resistance. AB - Cisplatin is a potent cytotoxic agent that functions as a bivalent electrophile, forming both interstrand and intrastrand DNA cross-links. Cisplatin-mediated DNA damage results in cell cycle arrest and initiation of apoptotic cell death. Increased cellular glutathione concentrations have been closely correlated with cisplatin resistance but do not reduce the extent of cisplatin-DNA adduct formation. One hypothesis to explain the ability of glutathione to inhibit cisplatin cytotoxicity is that glutathione, through its antioxidant function, plays a role in apoptotic regulatory pathways. We tested this hypothesis using MCF-7 breast cancer cells transfected with the apoptotic inhibitor Bcl-2. Bcl-2 overexpression in MCF-7 cells was associated with a nearly 3-fold increase in cellular glutathione levels and with increased resistance to cell death after cisplatin exposure. Treatment of MCF-7 lines with buthionine sulfoximine, an inhibitor of glutathione synthesis, normalized glutathione levels in Bcl-2 and control transfectants and completely abrogated Bcl-2-mediated cisplatin resistance without affecting Bcl-2 expression. Bcl-2 overexpression and up regulation of glutathione were not associated with a change in either cisplatin DNA adduct formation or repair over time. These results suggest that Bcl-2 mediated cisplatin resistance in MCF-7 cells is dependent on up-regulation of glutathione production, which contributes to cell survival by mechanisms independent of cisplatin inactivation or inhibition of DNA adduct formation. A similar dependence on glutathione for Bcl-2-mediated inhibition of cisplatin toxicity was confirmed in a second cell line, the lymphocytic precursor FL5.12. Taken together, these data suggest that apoptotic signaling after genotoxic exposure can be inhibited by the antioxidant activity of glutathione. Inhibition of glutathione synthesis or modulation of glutathione stores in tumors that overexpress Bcl-2 may comprise a novel anticancer strategy. PMID- 12543782 TI - Inducible activation of Ras and Raf in adult epidermis. AB - Ras effects vary with developmental setting, with oncogenic RAS activation implicated in epithelial carcinogenesis. In epidermal cells, previous studies described conflicting Ras impacts on growth and differentiation, with the only in vivo studies relying on constitutive alterations of Ras function throughout development. To study Ras effects in developmentally mature adult epidermis, we expressed a 4-hydroxytamoxifen (4OHT)-regulated Ras fusion in transgenic mice using the keratin 14 promoter. Resulting adult K14-ER:Ras mice displayed 4OHT inducible activation of Ras as well as elements of Raf/mitogen-activated protein kinase (MAPK) but not RalGDS/Ral or phosphatidylinositol 3'-kinase (PI3K)/Akt downstream Ras effector pathways. Ras reversibly induced massive cutaneous hyperplasia and suppressed differentiation. Ras-driven hyperproliferation was accompanied by increases in beta1 and beta4 integrin epidermal progenitor markers. Epidermal expression of inducible Raf produced similar changes. These findings indicate that activation of Ras in adult epidermis promotes proliferation and inhibits differentiation and that Raf is sufficient to mediate these effects. PMID- 12543783 TI - Characterization of a novel epigenetic effect of ionizing radiation: the death inducing effect. AB - The detrimental effects associated with exposure to ionizing radiation have long been thought to result from the direct targeting of the nucleus leading to DNA damage; however, the emergence of concepts such as radiation-induced genomic instability and bystander effects have challenged this dogma. After cellular exposure to ionizing radiation, we have isolated a number of clones of Chinese hamster-human hybrid GM10115 cells that demonstrate genomic instability as measured by chromosomal destabilization. These clones show dynamic and persistent generation of chromosomal rearrangements multiple generations after the original insult. We hypothesize that these unstable clones maintain this delayed instability phenotype by secreting factors into the culture medium. To test this hypothesis we transferred filtered medium from unstable cells to unirradiated GM10115 cells. No GM10115 cells were able to survive this medium. This phenomenon by which GM10115 cells die when cultured in medium from chromosomally unstable GM10115 clones is the death-inducing effect. Medium transfer experiments indicate that a factor or factors is/are secreted by unstable cells within 8 h of growth in fresh medium and result in cell killing within 24 h. These factors are stable at ambient temperature but do not survive heating or freezing, and are biologically active when diluted with fresh medium. We present the initial description and characterization of the death-inducing effect. This novel epigenetic effect of radiation has implications for radiation risk assessment and for health risks associated with radiation exposure. PMID- 12543784 TI - A novel human prostate-specific gene-1 (HPG-1): molecular cloning, sequencing, and its potential involvement in prostate carcinogenesis. AB - Prostate-specific genes that have a role in normal and abnormal prostate growth are needed for early and specific diagnosis and treatment of prostate cancer. In the present study, the differential display-PCR technique was used to obtain a prostate-specific approximately 339-bp cDNA fragment. On screening the human prostate lambdagt10 library with this fragment, a full-length 1468-bp human prostate-specific gene (HPG-1) with an open reading frame of 127 amino acids (aa) was retrieved. Extensive database search revealed that the HPG-1 has novel nucleotide/aa sequences. It was localized on Homo sapiens 3q26 chromosomal locus, a region that has been shown to be involved in prostate carcinoma. The computer generated translated protein has a calculated molecular mass of 14.8 kDa with several potential glycosylation and phosphorylation sites including two N-linked glycosylation, one tyrosine phosphorylation, and one N-myristoylation sites. The in vitro transcription and translation procedures using HPG-1 cDNA yielded a protein of similar molecular mass of approximately 15 kDa. Hydrophilicity analysis of the deduced aa sequence indicated that HPG-1 is a membrane anchored/attached protein. Analysis for tissue specificity by using the Northern blot and reverse transcription-PCR-Southern blot procedures using 19 different human tissues revealed that HPG-1 is expressed specifically only in prostate tissue. To examine its involvement in prostate carcinogenesis, three prostate cancer epithelial cell lines, one androgen-responsive (LNCaP) and two androgen nonresponsive (DU-145 and PC-3), were examined for the expression of HPG-1. Using the Northern blot and quantitative reverse transcription-PCR procedures it was found that LNCaP and DU-145 cells and not the PC-3 cells have HPG-1 expression, with LNCaP cells having approximately 2-3-fold higher levels of HPG-1 mRNA transcripts compared with DU-145 cells. In vitro culture of LNCaP cell with antisense and not the sense oligonucleotide decreased the HPG-1 mRNA levels and inhibited the cell growth in a concentration-dependent manner; at 72 h there was an 86% inhibition of cell growth. HPG-1 mRNA expression in LNCaP cells was found to be responsive to androgen. Thus, the novel androgen-responsive HPG-1, which has prostate-specific expression and seems to be involved in carcinogenesis, may have applications in the specific diagnosis and treatment of prostate cancer. PMID- 12543785 TI - The prognostic value of polymorphonuclear leukocyte elastase in patients with primary breast cancer. AB - A variety of serine proteases, including urokinase-type plasminogen activator (uPA), plasmin,and polymorphonuclear leukocyte elastase (PMN-E), have been implicated in the processes of tumor cell invasion and metastasis. Besides degrading of matrix proteins, PMN-E has been shown to be able to cleave and inactivate plasminogen activator inhibitor-1 (PAI-1), the main inhibitor of uPA, and alpha2-antiplasmin, the natural inhibitor of plasmin, thus enabling an uncontrolled matrix degradation by the fibrinolytic enzymes. Because only limited data are available on a relationship between the tumor level of PMN-E and prognosis in primary breast cancer patients, in the present study we have measured with an ELISA the levels of PMN-E (in complex with alpha1-proteinase inhibitor) in cytosolic extracts of 1143 primary breast tumors. Levels of complexed PMN-E have been correlated with the lengths of metastasis-free survival (MFS), relapse-free survival, and overall survival, and a comparison was made with data previously obtained for uPA and PAI-1. Our results show that patients with a high PMN-E level in their primary tumor had a rapid relapse and an early death compared with patients with a low tumor level of PMN-E. This held true for node-negative and node-positive subgroups of patients as well. The relationship of PMN-E with a poor prognosis was especially obvious during short-term follow-up (0-60 months). In Cox multivariate regression analysis, corrected for the traditional prognostic factors, PMN-E was an independent prognostic factor, and high levels of PMN-E were associated with a poor MFS [hazard ratio (HR), 1.63; 95% confidence interval (CI), 1.23-2.16; P < 0.001], relapse-free survival (HR, 1.45; 95% CI, 1.10-1.89; P = 0.01), and overall survival (HR, 1.64; 95% CI, 1.20 2.23; P = 0.003). Furthermore, in all three multivariate models, PMN-E still added significantly to the model after the additional inclusion of the uPA. PMN-E was an independent prognostic factor for MFS even in the multivariate analysis including the traditional clinical prognostic factors and the strong established biochemical prognostic factors uPA and PAI-1. Our present study suggests that PMN E is associated with breast cancer metastasis, and knowledge of the tumor PMN-E status might be helpful in selecting the appropriate individualized (adjuvant) treatment for patients with breast cancer. PMID- 12543786 TI - BRCA1 and BRCA2 mutation status and tumor characteristics in male breast cancer: a population-based study in Italy. AB - To investigate at the population level the impact of BRCA1/BRCA2 gene alterations in male breast cancer, we analyzed a population-based series of 25 male breast cancer cases from Florence, Central Italy. We combined mutational screening with the study of germ-line allele transcript levels and of tumor-associated losses of heterozygosity. Screening by protein truncation test and single-strand conformational polymorphism assay, followed by sequencing, revealed 4 pathogenetic mutations (4 of 25 = 16%; 95% confidence interval, 5-37%), 1 in BRCA1 and 3 in BRCA2, including mutations recurring in Central Italy (BRCA1 3345delAG and BRCA2 6696delTC). The a priori probability of carrying a mutation, estimated using BRCAPRO software, showed a good agreement between expected and observed mutations (14% versus 16%). A 7-fold association between germ-line mutations and family history of breast-ovarian cancer emerged. To investigate associations between BRCA1/BRCA2 status and clinicopathological characteristics, we analyzed the histopathological and immunophenotypic parameters of the tumors. A significant association emerged between mutation carrier status and high histological grade (P = 0.02). Furthermore, one BRCA2 carrier was affected with Paget's disease, an extremely rare male breast cancer histotype. Overall, BRCA1/2 mutations were observed to be strongly associated with positive c-erbB-2 immunostaining (P = 0.004). To evaluate germ-line allele expression, we used primer extension assays targeting frequent BRCA1 and BRCA2 polymorphisms. A BRCA2 allele transcript imbalance was found in one of four heterozygotes tested, all of them negative for germ-line mutations. BRCA1 transcript imbalances were not detected in nine heterozygotes analyzed. Losses of heterozygosity at one or more of nine loci in the BRCA2 region were found in 8 of 22 tumors tested. Interestingly, a case that was negative for BRCA1/BRCA2 germ-line mutations and that had a priori mutation probability <10% showed loss of heterozygosity at all three of the intragenic BRCA2 markers analyzed, which could be related to a somatic involvement of BRCA2. No losses of heterozygosity were detected at BRCA1. In conclusion, constitutional BRCA1/BRCA2 mutations accounted for 16% of the male breast cancer cases in this area of Central Italy. The detection of a BRCA2 germ line transcript imbalance and of a somatic loss of BRCA2 among the cases that resulted negative for germ-line mutations suggests a role of this gene more relevant than indicated by conventional mutational analysis. A distinct pattern of characteristics indicative of aggressive behavior, including high-grade and c erbB-2 expression, was evident in tumors from germ-line BRCA2 mutation carriers. This suggests that phenotypic characteristics may contribute to the identification of hereditary BRCA2-related male breast cancers and that these tumors might share a unique molecular pathway of cancer progression. PMID- 12543787 TI - Systemic reovirus therapy of metastatic cancer in immune-competent mice. AB - The human reovirus is an oncolytic virus that specifically targets cancer cells with an activated Ras pathway. Because it is replication competent and highly specific for cancer cells, this virus has the potential to be an effective antimetastatic cancer agent through remote site delivery. In this study, we exploited the ability of reovirus to replicate in murine cells to test the efficacy of this virus in eliminating distal and/or metastatic tumors in immune competent mice. We found that i.v. therapy with reovirus not only inhibited metastatic tumor growth but also led to a significant improvement in animal survival. Combining i.v. reovirus treatment with immune suppression (cyclosporine A or anti-CD4/anti-CD8 antibodies) resulted in further reduction in tumor size and a considerable prolongation in survival, compared with viral therapy alone. Combined therapy was also effective in overcoming a preexisting immunity to reovirus (a common occurrence in humans and thus a potential impediment to oncolytic effectiveness) to induce metastatic tumor regression. This is the first study to use systemic delivery of an oncolytic agent in conjunction with immune suppressive drugs to effectively prolong animal survival. Altogether, our results suggest that i.v. reovirus therapy may present a feasible, novel alternative in the treatment of metastatic cancer in humans. PMID- 12543788 TI - A universal pretargeting system for cancer detection and therapy using bispecific antibody. AB - Multistep targeting systems represent highly selective alternatives to targeting systems using directly radiolabeled antibodies for diagnostic and therapeutic applications. A flexible bispecific antibody (bsMAb) multistep, pretargeting system that potentially can be developed for use with a variety of different imaging or therapeutic agents is described herein. The flexibility of this system is based on use of an antibody directed against histamine-succinyl-glycine (HSG) and the development of peptides containing the HSG residue. HSG-containing peptides were synthesized with either 1,4,7,10-tetraazacyclododecane-N,N',N",N"' tetraacetic acid for the chelation of 111In, 90Y, or 177Lu, or a technetium/rhenium chelate. The peptides can be radiolabeled to a high specific activity in a facile manner that avoids the need for purification. In vivo studies in nude mice bearing human colon tumor xenografts showed that the radiolabeled peptides cleared rapidly from the body with minimal retention in tumor or normal tissues. For pretargeting, these peptides were used in combination with a bsMAb composed of the anti-HSG Fab' that was covalently coupled with the Fab' of either an anticarcinoembryonic antigen or an anticolon specific antigen-p antibody to provide tumor targeting capability. When the radiolabeled peptides were administered 1-2 days after a pretargeting dose of the bsMAbs, tumor uptake of the radiolabeled peptides increased as much as 28-175 fold over that seen with the peptides alone with tumor:nontumor ratios exceeding 2:1 to 8:1 within just 3 h of the peptide injection, which was a marked improvement over the tumor:nontumor ratios seen with a directly radiolabeled 99mTc-anti-anticarcinoembryonic antigen Fab' at this same time. The anticolon specific antigen-p x anti-HSG F(ab')2 bsMAb had the highest and longest retention in the tumor, and when used in combination with the 111In-labeled peptide, radiation dose estimates for therapeutic radionuclides, such as 90Y and 177Lu, suggested that antitumor effects would be expected with tolerable radiation exposure to the normal tissues. These results suggest that this multistep, pretargeting system has diagnostic imaging and therapeutic potential. PMID- 12543789 TI - Insulin-like growth factor I-mediated protection from rapamycin-induced apoptosis is independent of Ras-Erk1-Erk2 and phosphatidylinositol 3'-kinase-Akt signaling pathways. AB - The mTOR inhibitor rapamycin induces G1 cell cycle accumulation and p53 independent apoptosis of the human rhabdomyosarcoma cell line Rh1. Insulin-like growth factor I (IGF-I) and insulin, but not epidermal growth factor or platelet derived growth factor, completely prevented apoptosis of this cell line. Because the Ras-Erk1-Erk2 and phosphatidylinositol 3'-kinase (PI3K)-Akt pathways are implicated in the survival of various cancer cells, we determined whether protection from rapamycin-induced apoptosis by IGF-I requires one or both of these pathways. Despite the blocking of Ras-Erk signaling by the addition of PD 98059 (a MEK1 inhibitor) or by the overexpression of dominant-negative RasN17, IGF-I completely prevented rapamycin-induced death. Inhibition of Ras signaling did not prevent Akt activation by IGF-I. To determine the role of the PI3K-Akt pathway in rescuing cells from apoptosis caused by rapamycin, cells expressing dominant-negative Akt were tested. This mutant protein inhibited IGF-I-induced phosphorylation of Akt and blocked phosphorylation of glycogen synthase kinase 3. The prevention of rapamycin-induced apoptosis by IGF-I was not inhibited by expression of dominant-negative Akt either alone or under conditions in which LY 294002 inhibited PI3K signaling. Furthermore, IGF-I prevented rapamycin-induced apoptosis when the Ras-Erk1-Erk2 and PI3K-Akt pathways were blocked simultaneously. Similar experiments in a second rhabdomyosarcoma cell line, Rh30, using pharmacological inhibitors of PI3K or MEK1, alone or in combination, failed to block IGF-I rescue from rapamycin-induced apoptosis. Therefore, we conclude that a novel pathway(s) is responsible for the IGF-I-mediated protection against rapamycin-induced apoptosis in these rhabdomyosarcoma cells. PMID- 12543790 TI - SKI-606, a 4-anilino-3-quinolinecarbonitrile dual inhibitor of Src and Abl kinases, is a potent antiproliferative agent against chronic myelogenous leukemia cells in culture and causes regression of K562 xenografts in nude mice. AB - Constitutive tyrosine kinase activity of Bcr-Abl promotes proliferation and survival of chronic myelogenous leukemia (CML) cells. Inhibition of Bcr-Abl tyrosine kinase activity or signaling proteins activated by Bcr-Abl in CML cells blocks proliferation and causes apoptotic cell death. The selective Abl kinase inhibitor, STI-571 (marketed as Gleevec), is toxic to CML cells in culture, causes regression of CML tumors in nude mice, and is currently used to treat CML patients. Here we describe a p.o. active, dual Src/Abl kinase inhibitor with potent antiproliferative activity against CML cells in culture. This 4-anilino-3 quinolinecarbonitrile (SKI-606) ablates tyrosine phosphorylation of Bcr-Abl in CML cells and of v-Abl expressed in fibroblasts. SKI-606 inhibits phosphorylation of cellular proteins, including STAT5, at concentrations that inhibit proliferation in CML cells. Phosphorylation of the autoactivation site of the Src family kinases Lyn and/or Hck is also reduced by treatment with SKI-606. Once daily oral administration of this compound at 100 mg/kg for 5 days causes complete regression of large K562 xenografts in nude mice. PMID- 12543791 TI - The designed angiostatic peptide anginex synergistically improves chemotherapy and antiangiogenesis therapy with angiostatin. AB - Recently, we demonstrated that the designed peptide anginex displays potent antiangiogenic activity. The aim of the present study was to investigate anginex treatment as a single-agent therapy and to test its ability to improve conventional chemotherapy and antiangiogenesis therapy. In a human ovarian carcinoma mouse model, anginex inhibited tumor growth by 70%. When anginex was combined with a suboptimal dose of carboplatin, tumors regressed to an impalpable state. Anginex plus angiostatin worked synergistically to inhibit tumor growth. Assessment of microvessel density suggested that the antitumor activity of anginex is mediated by angiogenesis inhibition. In any of the experiments, no sign of anginex-induced toxicity was observed. PMID- 12543792 TI - Antitumor activity of the 16-kDa prolactin fragment in prostate cancer. AB - The 16-kDa prolactin (PRL), derived from the proteolytic cleavage of wild-type 23 kDa PRL, has been shown to have antiangiogenic activity. Such an antiangiogenic activity may have an effect on tumor growth in vivo. Here we examined the effect of 23-kDa and 16-kDa PRL on tumor growth, and the potential of using recombinant 16-kDa human PRL for prostate cancer therapy. The effects of 23-kDa PRL and 16 kDa PRL on the tumorigenicity of prostate cancer cells in vivo were studied. Using an adenovirus transfer vector to achieve high efficiency 23-kDa and 16-kDa PRL transfection in DU145 and PC-3 human prostate carcinoma cell lines, we demonstrated that expression of 16-kDa PRL in the prostate cancer cells markedly reduced their ability to form tumors in a xenograft animal model. These studies established that the 16-kDa PRL has antitumor activity in vivo, presumably as a result of its antiangiogenic effect. Interestingly, 23-kDa PRL showed a weak and transient suppression of prostate tumor growth. The weak antitumor activity of 23 kDa PRL may be because of the production of 16-kDa PRL from 23-kDa PRL by the tumor cells. Thus, the apparent effect of 23-kDa PRL on the growth of DU145 and PC-3 cells in vivo may result from the combined effects of 23-kDa PRL and 16-kDa PRL. These results suggest that the 16-kDa PRL has potential as a treatment agent in prostate cancer. PMID- 12543793 TI - CpG oligonucleotides enhance the tumor antigen-specific immune response of a granulocyte macrophage colony-stimulating factor-based vaccine strategy in neuroblastoma. AB - Granulocyte macrophage colony-stimulating factor (GM-CSF)-transduced autologous tumor cells form the basis of many immunotherapeutic strategies. We tested whether combining this approach with T-helper 1 (Th-1)-like immunostimulatory CpG oligodeoxynucleotides (CpG ODNs) would improve therapeutic efficacy in an established model of murine neuroblastoma. The weakly immunogenic Neuro-2a cell line was used in syngeneic A/J mice. CpG 1826 was tested for its antitumor effect alone and as an adjuvant to Neuro-2a cells retrovirally transduced to express murine GM-CSF (GM/Neuro-2a). Three days after wild-type (WT) tumor cell inoculation, mice in different groups were s.c. vaccinated in the opposite leg with combinations of WT neuro2a, irradiated (15 Gy) WT or GM/Neuro-2a transfectants with or without CpG 1826 (200 micro g). To test for the induction of memory responses, mice that rejected their tumor were rechallenged with WT Neuro-2a (1 x 10(6)) 7 weeks after vaccination. All of the mice in the control (unvaccinated) group died within 3 weeks after Neuro-2a inoculation. Most of the vaccinated groups had only minimal-to-modest antitumor responses, and the mice succumbed to tumor. Tumor growth was remarkably inhibited in the group of mice that received irradiated GM/Neuro-2a plus CpG and four (50%) of eight mice in this group survived tumor free. Tumor-free mice were resistant to further WT tumor cell challenge, indicating a memory response. Mechanistic studies showed that CpG alone induced a favorable Th-1-like cytokine immune response and vaccine induced tumor cell killing was dependent on both CD4 and CD8 T cells that killed tumor cell targets by apoptosis. These results demonstrate that CpG ODNs enhanced the antitumor effect of irradiated GM-CSF secreting Neuro-2a cells. This vaccine strategy elicits a potent tumor antigen-specific immune response against established murine neuroblastoma and generates systemic neuroblastoma-specific immunity. PMID- 12543794 TI - Association between human leukocyte antigen polymorphism and human papillomavirus 16-positive vulval intraepithelial neoplasia in British women. AB - Polymorphisms in human leukocyte antigen (HLA) genes have been implicated in the risk for developing human papillomavirus (HPV)-associated cervical neoplasia. By comparison with local cadaver controls typed for HLA class I (n = 946) and II (n = 144) antigens, HPV-16-positive high grade vulval intraepithelial neoplasia patients (n = 42) showed significantly different frequencies of HLA-A2 [odds ratio (OR), 2.1; confidence interval (CI), 1.4-3.9], HLA-B7 (OR, 2.6; CI, 1.4 4.7), HLA-DRB1*01(01/02/04) (OR, 0.1; CI, 0.03-0.5), HLA-DRB1*11 (OR, 3.3; CI, 1.4-7.1), HLA-DRB1*13 (OR, 0), HLA-DQB1*05 (OR, 0.2; CI, 0.05-0.6), and HLA DQB1*03032 (OR, 4.6; CI, 1.5-14.0). With the exception of HLA-B7 and HLA-DRB1*11, these significant differences were also seen comparative to local HPV-16-positive cervical carcinoma patients (n = 114), suggesting a specific immunogenetic contribution that is independent of HPV-16 infection in high-grade vulval intraepithelial neoplasia. Such factors are important to the development of HPV vaccines for treatment of cervical and vulval neoplasia. PMID- 12543795 TI - An integrated physical and gene map of the 3.5-Mb chromosome 3p21.3 (AP20) region implicated in major human epithelial malignancies. AB - To facilitate the identification of tumor suppressor genes in the chromosome 3p21.3-p22 AP20 subregion, we constructed a 3.5-Mb physical and gene map of this segment (between markers D3S4285 and D3S3873) that spans the distance from 124.4cR3000 to 133.5 cR3000 of the GB4 genetic map. We used NotI-linking and jumping clones, sequence-tagged site PCR marker analysis, and multicolor and fiber fluorescence in situ hybridization to confirm the sequence order and map orientation. An integrated clone contig composed of 5 yeast artificial chromosome, 15 bacterial artificial chromosome, 5 P1 artificial chromosome, and 8 NotI-linking clones provided the physical base of the map. We unequivocally established the order of 28 sequence-tagged sites and 35 genes in the region. Gaps between published bacterial artificial chromosome contigs were determined and covered by our own sequence data. Furthermore, three new genes were isolated, namely the human homologue to the rat Golgi peripheral membrane protein p65, GOLPH5 (GORASP1), the gene for stress-inducible protein, STI2, and the AP20 region gene 1, APRG1. The tumor suppressor gene candidate APRG1 was positioned close to the border of the homozygous deletion in a small cell lung cancer cell line ACC-LC5. Expression analysis with a tissue-specific panel of cDNA revealed seven distinct tissue-specific splice variants (A-G) of the message (size range, 1.0-1.8 kb). Although the gene was expressed at a low level in all tested tissues, comparatively higher expression was detected in pancreas (splice forms B and D), kidney (A) and placenta (B and C). The APRG1 gene encoded a predicted protein of 170 amino acids (isoform B), which had an NH2-terminal part conserved among members of the eukaryotic translation factor 6 gene family. A Prosite pattern corresponding to the cell attachment sequence Arg-Gly-Asp was also found. The presence of this domain raised the intriguing possibility that APRG1B may be directly involved in membrane interactions and cell adhesion. We showed that the AP20 region was duplicated during mammalian evolution and homologous gene clusters were present in human chromosome 2 and syntenic mouse regions on chromosomes 1, 2, and 9. Interestingly, the HYA22 gene (human ortholog of the yeast YA22 gene) was located at the borders of both breakpoints, evolutionarily conserved gene cluster and homozygous deletions detected in lung, kidney and other cancers. NotI digestion revealed that the AP20 region was frequently and extensively methylated in renal carcinoma cell lines and tumor biopsies. PMID- 12543796 TI - Selection pressures of TP53 mutation and microenvironmental location influence epidermal growth factor receptor gene amplification in human glioblastomas. AB - Epidermal growth factor receptor (EGFR) gene amplification occurs in glioblastomas as so-called double minutes. Because double minutes are extrachromosomal fragments, selection pressures must operate to maintain high EGFR copy number over multiple cell divisions. In analyses of glioblastoma lysates, EGFR amplification has been observed almost exclusively in glioblastomas harboring wild-type TP53 genes, which raises the alternative hypotheses that TP53 mutation either prevents amplification or selects against maintenance of EGFR amplified cells. To address these possibilities at the cellular level, we studied 14 glioblastomas for TP53 mutation and EGFR gene amplification status, using fluorescence in situ hybridization (FISH) for the latter. Remarkably, four of the six cases with TP53 mutation had isolated EGFR-amplified cells in different regions, demonstrating that EGFR amplification occurs frequently at the cellular level in TP53-mutant glioblastomas. Thus, TP53 mutation does not prevent EGFR amplification but does not facilitate selection of EGFR-amplified cells. Of the eight cases without TP53 mutation, five had widespread EGFR amplification. In four of these five cases, multiple regions of the tumor were available for examination; FISH demonstrated a gradation of EGFR amplification, with highly amplified cells, primarily at the invading edges rather than the relatively solid tumor centers, suggesting that EGFR overexpression, when selected for in vivo, may be related to tumor invasion. PMID- 12543797 TI - BRCA1/2 mutation status influences somatic genetic progression in inherited and sporadic epithelial ovarian cancer cases. AB - Metaphase comparative genomic hybridization was used to analyze the spectrum of genetic alterations in 141 epithelial ovarian cancers from BRCA1 and BRCA2 mutation carriers, individuals with familial non-BRCA1/2 epithelial ovarian cancer, and women with nonfamilial epithelial ovarian cancer. Multiple genetic alterations were identified in almost all tumors. The high frequency with which some alterations were identified suggests the location of genes that are commonly altered during ovarian tumor development. In multiple chromosome regions, there were significant differences in alteration frequency between the four tumor types suggesting that BRCA1/2 mutation status and a family history of ovarian cancer influences the somatic genetic pathway of ovarian cancer progression. These findings were supported by hierarchical cluster analysis, which identified genetic events that tend to occur together during tumorigenesis and several alterations that were specific to tumors of a particular type. In addition, some genetic alterations were strongly associated with differences in tumor differentiation and disease stage. Taken together, these data provide molecular genetic evidence to support previous findings from histopathological studies, which suggest that clinical features of ovarian and breast tumors differ with respect to BRCA1/2 mutation status and/or cancer family history. PMID- 12543798 TI - Overexpression of cyclin D1 contributes to malignancy by up-regulation of fibroblast growth factor receptor 1 via the pRB/E2F pathway. AB - Overexpression of cyclin D1 due to gene rearrangement, gene amplification, or simply increased transcription occurs frequently in several types of human cancers. However, overexpression of cyclin D1 in cell culture system is insufficient, by itself, to cause malignant transformation. In the present study, we found that when rodent fibroblasts that overexpress cyclin D1, but not normal fibroblasts, were treated with basic fibroblast growth factor (bFGF), there was enhanced cell cycle progression, extracellular signal-regulated kinase 2 activation, induction of anchorage-independent growth, and enhanced invasion of a Matrigel barrier. These enhanced responses to bFGF appear to be due to increased expression of fibroblast growth factor receptor 1, at both the mRNA and protein levels, in the cyclin D1-overexpressing cells. We obtained evidence that this increase in fibroblast growth factor receptor 1 expression is mediated through cyclin D1 activation of the pRB/E2F pathway. Taken together, these results suggest that in vivo cyclin D1 overexpression can enhance tumor progression, at least in part, by potentiating the stimulatory efforts of bFGF, which is often produced by stromal cells, and the growth of adjacent tumor cells. PMID- 12543799 TI - Melanoma metastasis suppression by chromosome 6: evidence for a pathway regulated by CRSP3 and TXNIP. AB - Loss of genetic material on chromosome 6 has been associated with progression of human melanomas. We showed previously that introducing chromosome 6 into metastatic human melanoma cell lines suppresses metastasis without affecting the ability of the hybrids to form progressively growing tumors. By subtractive hybridization comparing nonmetastatic chromosome 6-containing (neo6/C8161) versus parental (C8161) metastatic cells, the KISS1 metastasis suppressor gene was isolated. However, KISS1 mapped to chromosome 1q32. To identify upstream regulator(s) of (and downstream effectors of) KISS1, microarray hybridization comparing C8161 and neo6/C8161 variants was performed. TXNIP/VDUP1, a thioredoxin binding protein, was expressed more highly in neo6/C8161 and in nonmetastatic melanomas. Increased TXNIP expression inhibited metastasis and up-regulated KISS1. Surprisingly, TXNIP also mapped to chromosome 1q. PCR karyotyping that refined the region on chromosome 6 identified CRSP3/DRIP130, a transcriptional coactivator, as a metastasis suppressor. CRSP3 transfectant cells had up regulated KISS1 and TXNIP expression and were suppressed for metastasis. Quantitative real-time reverse-transcription PCR of clinical melanoma samples showed that loss of CRSP3 expression correlated with decreased KISS1 expression and increased metastasis. Thus, we implicated a specific gene on chromosome 6 in the etiology of melanoma metastasis and identified potential up-stream regulators of KISS1 and TXNIP. PMID- 12543800 TI - Expression of differentiation melanoma-associated antigen genes is associated with favorable disease outcome in advanced-stage melanomas. AB - Cutaneous melanomas have been found to express several immunogenic differentiation melanoma-associated antigens (MAAs) that have been suggested to play an important role in disease outcome. Adaptive host immunity to MAAs has shown some level of control on melanoma progression. To date, there has been no definitive report correlating the level of differentiated MAAs gene expression in melanomas with overall disease outcome. Metastasis of melanoma to distant visceral organ sites usually indicates a survival of less than 1 year; however, a subset of patients who undergo cytoreductive surgery of distant metastases survive for a longer period. We hypothesized that the gene expression level of differentiation MAAs in metastatic melanoma (AJCC stage IV) lesions would be predictive of survival. We focused on three known differentiation MAAs: tyrosinase (TYR), TYR-related protein 2 (TRP-2), and melanoma antigen recognized by T cells 1 (MART-1); all three of them are known to induce immune responses in melanoma patients and are frequently expressed in melanomas. A quantitative reverse-transcriptase RealTime PCR (qRT) assay was developed for these MAAs to assess mRNA expression in metastatic melanoma tumors obtained from cytoreductive surgery of AJCC stage IV melanoma patients (n = 35). Patients were followed up for over 60 months. There was a variation in mRNA copy levels for individual MAAs in melanoma tumors. Elevated MAA mRNA copy levels of TYR and TRP-2 significantly (P < 0.03 and < 0.009, respectively) correlated with improved overall survival. Patients having at least one MAA expressed in their tumors had a significantly (P = 0.01) better overall survival (median 16 months). These studies demonstrate that levels of differentiated MAA mRNA expression of advanced-stage metastatic melanomas can be used as molecular predictive factors of disease outcome. The studies also imply that an assessment of melanoma tumor MAAs may provide a stratification factor targeted for active-specific immunotherapy. PMID- 12543801 TI - The homeotic protein Six3 is a coactivator of the nuclear receptor NOR-1 and a corepressor of the fusion protein EWS/NOR-1 in human extraskeletal myxoid chondrosarcomas. AB - Nuclear receptors represent a large family of transcription factors involved in development, differentiation, homeostasis, and cancer. In recent years, a growing number of cofactors has been discovered that participate in the regulation of the transcriptional activity of these proteins. We present in this study the identification of a cofactor, the homeotic protein Six3, which differentially regulates the transcriptional activity of the orphan nuclear receptor NOR-1 (NR4A3). NOR-1 is normally involved in the balance between cell proliferation and cell death, and is implicated in oncogenesis as part of the EWS/NOR-1 fusion protein found in human extraskeletal myxoid chondrosarcoma (EMC) tumors. Reverse transcription-PCR analyses indicate that EMC tumors expressing the EWS/NOR-1 mRNA also express mRNAs encoding NOR-1 and Six3. Glutathione S-transferase fusion protein assays show that Six3 binds in vitro the DNA-binding domain of NOR-1 and the EWS domain of EWS/NOR-1 and that the homeodomain of Six3 is required for these interactions. Mammalian two-hybrid experiments, using immortalized human chondrocytes as a model, indicate that Six3 also interacts with NOR-1 and EWS/NOR 1 in vivo. Cotransfection experiments show that Six3 stimulates the transcriptional activity of NOR-1, whereas it represses that of EWS/NOR-1. Considering the highly specific expression pattern of Six3, our finding that it is expressed in EMC suggests that it plays a pivotal role in the development of these tumors. We propose that Six3 maintains a transcriptional balance between the activities of NOR-1 and EWS/NOR-1, the net effect being to deregulate the expression of specific target genes and push the equilibrium toward uncontrolled cell proliferation. PMID- 12543802 TI - Deletion of chromosome 3p14.2-p25 involving the VHL and FHIT genes in conventional renal cell carcinoma. AB - Loss of heterozygosity (LOH) at chromosome 3p and inactivation of the VHL gene are associated with the development of conventional renal cell carcinomas (RCCs). Recently, it was suggested that LOH at the FHIT gene at 3p14.2 is an early event in the development of RCC and is characteristic for all types of RCC. We have analyzed 88 conventional, 30 papillary, and 22 chromophobe RCCs for LOH at the VHL and FHIT regions and at other loci on chromosome 3p. A continuous deletion of 3p14.2-p25 harboring the VHL and FHIT genes occurred in 96% of the conventional RCCs but only in 10% of the papillary RCCs and 18% of the chromophobe RCCs. Our data indicate that LOH at chromosome 3p14.2-p25 is specific for conventional RCC and that loss of one allele of both the VHL and FHIT genes occurs in early stage of tumorigenesis. PMID- 12543803 TI - High levels of telomere dysfunction bestow a selective disadvantage during the progression of human oral squamous cell carcinoma. AB - Human epithelial cells experience multiple barriers to cellular immortality in culture (mortality mechanisms 0, 1, and 2). Mortality mechanism 2 (M2) is termed crisis and involves telomere dysfunction due to lack of telomerase. However, proliferating normal keratinocytes in vivo can express telomerase, so it is unclear whether human squamous cell carcinomas (SCCs), which usually have high telomerase levels, develop from preexisting telomerase-positive precursors or by the activation of telomerase in telomerase-deficient somatic cells. We show that 6 of 29 oral SCCs show characteristics of M2 crisis in vivo, as indicated by a high anaphase bridge index (ABI), which is a good correlate of telomere dysfunction, and that 25 of 29 tumors possess some anaphase bridges. ABIs in excess of 0.2 in the primary tumor showed a decrease in the corresponding lymph node metastases. This suggests that high levels of telomere dysfunction (>0.2) and, by inference, M2 crisis bestow a selective disadvantage on SCCs during progression stages of the disease. Supporting this, SCCs with high levels of telomere dysfunction grow poorly in culture, and the ectopic expression of telomerase corrects this, together with other features of M2 crisis. Our data suggest that a substantial proportion of oral SCCs in vivo ultimately arise from telomerase-deficient keratinocytes rather than putative telomerase-proficient cells in the undifferentiated parts of the epithelium. Furthermore, the presence of significant levels of telomere dysfunction in a high proportion of SCCs at diagnosis but not in the normal epithelium implies that the therapeutic inhibition of telomerase should selectively compromise the growth of such tumors. PMID- 12543804 TI - 2-methoxyestradiol up-regulates death receptor 5 and induces apoptosis through activation of the extrinsic pathway. AB - 2-Methoxyestradiol (2ME2), a natural metabolite of estradiol, is a potent antitumor and antiangiogenic agent. In vitro, 2ME2 inhibits the proliferation of a wide variety of cell lines and primary cultures, and in numerous models in vivo, it has been shown to be an effective inhibitor of tumor growth and angiogenesis. 2ME2 is currently in several Phase I and Phase II clinical trials under the name Panzem. Although various molecular targets have been proposed for this compound, the mechanism by which 2ME2 exerts its effects is still uncertain. This study shows that 2ME2 uses the extrinsic pathway for induction of apoptosis. 2ME2 treatment results in up-regulation of death receptor 5 (DR5) protein expression in vitro and in vivo and renders cells more sensitive to the cytotoxic activities of the DR5 ligand tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). 2ME2-induced apoptosis requires caspase activation and kinetic studies show the sequential activation of caspase-8, caspase-9, and caspase-3. Blockage of death receptor signaling by expression of dominant-negative Fas associated death domain severely attenuates the ability of 2ME2 to induce apoptosis. Because 2ME2 administration has not manifested dose-limiting toxicity in the clinic, DR5 expression may serve as a surrogate marker for biological response. PMID- 12543806 TI - A nuclear form of the heparin-binding epidermal growth factor-like growth factor precursor is a feature of aggressive transitional cell carcinoma. AB - Heparin-binding epidermal growth factor-like growth factor (HB-EGF), a member of the ErbB receptor ligand family, exists in distinct molecular forms with disparate biological activities. Previous studies have shown that the HB-EGF precursor, proHB-EGF, localizes to the cytoplasm of transitional cells of the human bladder urothelium and that the soluble form of the growth factor is an autocrine urothelial cell mitogen. In this study, we identify a potential role for proHB-EGF in transitional cell carcinoma (TCC) of the bladder. In an analysis of 33 TCC specimens and 8 normal controls, proHB-EGF, identified using an antibody directed against the cytoplasmic tail domain, localized to cell nuclei in a manner that correlated positively with tumor stage and grade (P < 0.001). The ability of proHB-EGF to localize to the nucleus was independently confirmed in a TCC cell line (TCCSUP), in which approximately 40% of transfected proHB-EGF was found to reside in the nuclear compartment. In Kaplan-Meier survival analysis, TCC patients with >20% proHB-EGF-positive cell nuclei demonstrated markedly reduced survival compared with patients with <20% proHB-EGF-positive nuclei (P < 0.005, log-rank test). In multivariate analysis, nuclear localization of proHB-EGF of >20% was an independent prognostic indicator of disease-specific mortality. This is the first report in any cell type that HB-EGF is capable of translocating to the cell nucleus. In addition, our findings suggest that nuclear proHB-EGF may play a role in disease progression in bladder cancer and possibly other cancers. PMID- 12543805 TI - Abrogation of the retinoblastoma tumor suppressor checkpoint during keratinocyte immortalization is not sufficient for induction of centrosome-mediated genomic instability. AB - Deregulation of the retinoblastoma (pRB) tumor suppressor pathway and telomerase activation have been identified as rate-limiting steps for immortalization of primary human epithelial cells. However, additional molecular aberrations including p53 inactivation, ras activation, and deregulation of protein phosphatase 2A activity are necessary for full transformation of immortalized epithelial cells. Genomic instability is observed in most human tumors and constitutes an important mechanism to allow emerging tumor cells to acquire additional mutations to efficiently overcome selection barriers during carcinogenic progression. In an attempt to model oral cancer in a human cell based system, we analyzed normal oral epithelial keratinocytes with the pRB pathway dysregulated by loss of expression of the cyclin-dependent kinase (cdk) 4/cdk6 inhibitor p16(INK4A) and/or ectopic expression of cdk4 or expression of the human papillomavirus (HPV) type 16 E7 oncoprotein. Ectopic expression of cdk4 and HPV-16 E7 was equally efficient in extending the life span of normal oral keratinocytes, and each was able to cooperate with telomerase (hTERT) to immortalize these cells. HPV-16 E7/hTERT-immortalized normal oral keratinocytes showed centrosome abnormalities, whereas populations of cdk4/hTERT-immortalized cells or hTERT-immortalized cells that had lost expression of p16INK4A showed no such abnormalities. These results demonstrate that disruption of the p16INK4A/pRB checkpoint of epithelial cell immortalization does not necessarily lead to centrosome-associated genomic instability. PMID- 12543807 TI - Interleukin-18 binding protein reduces b16 melanoma hepatic metastasis by neutralizing adhesiveness and growth factors of sinusoidal endothelium. AB - We studied the role of endogenous interleukin (IL)-18 in hepatic metastasis by blocking this cytokine using the naturally occurring IL-18 binding protein (IL 18BP). A single i.p. dose of IL-18BP given 30 min before intrasplenic injection of murine B16 melanoma (B16M) cells reduced the number of hepatic metastatic foci by 75% and metastatic volume by 80%. Same treatment reduced the intrahepatic retention of luciferase-transfected B16M by 50% and abolished VCAM-1 up regulation in the hepatic microvasculature, as assessed by reverse transcription PCR, Western blot, and immunohistochemistry. Twelve hours after IL-18BP, hepatic sinusoidal endothelium (HSE) cells were isolated, and adhesion of B16M cells to these cultured HSE cells was reduced to the level of vehicle-treated mice. IL 18BP treatment of mice with established micrometastases resulted in a 25% decrease in metastasis number and 40% decrease in metastasis volume, suggesting inhibition of endogenous growth factors. Indeed, the addition of IL-18BP to normal HSE abolished the release of melanoma cell growth factor(s) induced by B16M. IL-18 promoted the in vitro growth of B16M and human melanoma cells, which was IL-1 dependent. These data demonstrate a significant role of endogenous IL-18 on hepatic metastasis by up-regulating melanoma cell adhesion to HSE cells and tumor growth, implicating a possible antimetastatic benefit of neutralizing IL 18. PMID- 12543808 TI - Methyl-CpG-binding domain protein-2 mediates transcriptional repression associated with hypermethylated GSTP1 CpG islands in MCF-7 breast cancer cells. AB - GSTP1, encoding the pi-class glutathione S-transferase, is commonly inactivated by somatic CpGisland hypermethylation in cancers of the prostate, liver, and breast. We report here thathypermethylation of CpG dinucleotides at the 5' transcriptional regulatory region was sufficient to inhibit GSTP1 transcription in MCF-7 breast cancer cells and that repression of GSTP1 transcription was mediated in part by the methyl-CpG-binding domain (MBD) protein MBD2. MCF-7 breast cancer cells contained only hypermethylated GSTP1 CpG island alleles and failed to express GSTP1 mRNA or GSTP1 polypeptides. In contrast, MCF-7/ADR cells contained only unmethylated GSTP1 CpG island alleles and exhibited abundant GSTP1 expression. Chromatin immunoprecipitation analysis detected the presence of MBD2 and DNMT1 at the GSTP1 promoter in MCF-7 breast cancer cells but not in MCF-7/ADR breast cancer cells. In a test of the contribution of MBD2 to GSTP1 repression in MCF-7 breast cancer cells, transfection of small interference RNA complementary to MBD2 mRNA into MCF-7 cells both reduced MBD2 polypeptide levels and stimulated GSTP1 mRNA expression. These findings implicate MBD2 in GSTP1 silencing associated with somatic GSTP1 CpG island hypermethylation in breast cancer cells. PMID- 12543809 TI - The aldo-keto reductase AKR1C3 is a novel suppressor of cell differentiation that provides a plausible target for the non-cyclooxygenase-dependent antineoplastic actions of nonsteroidal anti-inflammatory drugs. AB - We and others have demonstrated expression of the aldo-keto reductase AKR1C3 in myeloid leukemia cell lines and that inhibitors of the enzyme, including nonsteroidal anti-inflammatory drugs (NSAIDs), promote HL-60 differentiation in response to all-trans retinoic acid (ATRA) and 1alpha,25-dihydroxyvitamin D3 (D3). Here, we demonstrate that overexpression of AKR1C3 reciprocally desensitizes HL-60 cells to ATRA and D3, thus confirming the enzyme as a novel regulator of cell differentiation. AKR1C3 possesses marked 11-ketoreductase activity converting prostaglandin (PG) D2 to PGF2alpha. Supplementing HL-60 cultures with PGD2 mimicked treatment with AKR1C3-inhibitors by enhancing the differentiation of the cells in response to ATRA. However, PGD2 is chemically unstable, being converted first to PGJ2 and then stepwise to 15-deoxy Delta(12,14)-prostaglandin J2(15Delta-PGJ2), a natural ligand for the peroxisome proliferator-activated receptor-gamma (PPARgamma). Consistent with this, PGD2 was rapidly converted to PGJ2 under normal tissue culture conditions but not in the presence of recombinant AKR1C3 when PGF2alpha was predominantly formed. In addition, PGJ2 but not PGF2alpha recapitulated the potentiation of HL-60 differentiation by PGD2 and AKR1C3 inhibitors. Furthermore, the capacity of all of these treatments to potentiate HL-60 cell differentiation was significantly reduced in the presence of the PPARgamma-antagonist GW 9662. We conclude that AKRIC3 protects HL-60 cells against ATRA and D3-induced cell differentiation by limiting the production of natural PPARgamma ligands via the diversion of PGD2 toward PGF2alpha and away from PGJ2. In addition, these observations identify AKR1C3 as plausible target for the non-cyclooxygenase-dependent antineoplastic actions of NSAIDs. PMID- 12543810 TI - Failure of activation of caspase-9 induces a higher threshold for apoptosis and cisplatin resistance in testicular cancer. AB - Testicular germ cell cancer is one of the very few cancers that are highly sensitive to and curable by cisplatin-based chemotherapy even in an advanced stage. However, in a few cases resistance to cisplatin occurs and patients subsequently die from progressive disease. The molecular basis for this resistance remains to be determined. Using two cisplatin-sensitive (2102EP and H12.1) and one cisplatin-resistant human testicular germ cell cancer cell line (1411HP), we investigated molecular mechanisms in the induction of apoptosis after cisplatin-treatment focusing on the cleavage and activation of caspase-2, caspase-3, caspase-7, caspase-8, and caspase-9. The cell line 1411HP showed a 3.3 fold cisplatin resistance when compared with the sensitive cell lines 2102EP and H12.1 by IC(90)s, which was treatment schedule independent (2- or 24-h incubation). Cisplatin resistance was associated with substantially decreased apoptosis in vitro and in derived nude mice xenografts as determined by Apo 2.7 detection, DNA-laddering, immunohistochemistry of active caspase-3, and terminal deoxynucleotidyl transferase-mediated nick end labeling assay. Total DNA platination as assessed by ELISA after cisplatin treatment in equimolar doses did not differ between cisplatin-resistant or -sensitive cells. In separate analysis of cells of early and late apoptotic stages, initiation of cisplatin-induced apoptosis appeared to be rather mediated by caspase-9 than by caspase-8. Resistant 1411HP cells failed to activate caspase-9 during the induction of apoptosis after cisplatin treatment at the IC(90) dose. Interestingly, inhibition of caspase-9 in sensitive H12.1 almost completely blocked apoptosis and induced cisplatin resistance to the same extent as in 1411HP so that apoptosis could only be induced by 3.3-fold higher cisplatin doses. Furthermore, in caspase-9 blocked cells, initiation of apoptosis occurred in a caspase-9 independent manner accompanied by activation of caspase-2 and caspase-3, which are intrinsic characteristics of resistant 1411HP cells. Failure of caspase-9 activation and cisplatin resistance was independent of the expression of p53, Bcl-2 family proteins, Fas receptor, and Fas ligand. In conclusion, failure of activation of the caspase-9 pathway induces a higher cellular threshold for cisplatin-mediated induction of apoptosis in testicular cancer cells. However, this higher threshold can be overcome by higher cisplatin doses, conceivably by using an alternate, caspase-9-independent apoptotic pathway. This supports the current clinical strategy of high-dose chemotherapy in patients with chemorefractory germ cell tumors. However, additional defining and eventually targeting the exact molecular mechanism blocking caspase-9 activation might lead to more selective therapeutic approaches to overcome cisplatin resistance in germ cell cancer. PMID- 12543811 TI - Cyclooxygenase-2 expression and prostanoid biogenesis reflect clinical phenotype in human colorectal fibroblast strains. AB - Up-regulated cyclooxygenase (COX)-2 expression and prostaglandin E2 (PGE2)synthesis contribute causally to the early stages of colorectal neoplasia and carcinogenesis, yet COX-2 expression is barely detectable in normal and premalignant colorectal epithelium. Rather, COX-2 expression in nonmalignant colonic tissue is probably confined to subepithelial cells, such as fibroblasts. We established a panel of 33 primary subepithelial fibroblast strains from human colonoscopic biopsies of normal colon (group I), normal segments of colons that harbored synchronous advanced neoplasms in remote segments (group II), advanced neoplasms (group III), and segments of active ulcerative colitis (group IV). In group I strains, mean basal and peak PGE2 levels after 24 h of interleukin (IL) 1beta stimulation were 5.4 +/- 1.1 and 32.8 +/- 4.9 ng/mg protein, respectively. Mean IL-1beta-stimulated peak levels in groups II, III, and IV strains were, respectively, 6-, 9-, and 7-fold greater than that in group I (P < 0.001 for each comparison), and inductions of COX-2 mRNA and protein were consistent with these findings. IL-1beta-mediated stimulation of PGE2 was fully blocked in the presence of a nonselective COX inhibitor (indomethacin) or a selective COX-2 inhibitor (NS 398). IL-1beta treatment elicited from group I (normal) and group III (cancer associated) fibroblasts, respectively, approximately 2- and 3-fold inductions of COX-2 transcriptional activity and approximately 1.4- and 1.7-fold inductions of COX-2 promoter activity. This modestly greater COX-2 transcription rate could not alone account for the dramatically higher levels of COX-2 mRNA and protein and PGE2 in cancer-associated compared with normal fibroblasts. However, incubation of fibroblasts with PGE2 after IL-1beta stimulation prolonged COX-2 mRNA half life from approximately 1 to 9 h. Our results strengthen the evidence that fibroblasts and other mesenchymal cells are the source of COX-2 expression in normal and premalignant colorectal tissue. Group II fibroblasts from normal segments of colons that harbored synchronous remote advanced neoplasms behaved like group III fibroblasts from advanced neoplasms and not group I fibroblasts from normal colons. We hypothesize that the effects of modestly greater COX-2 transcription in groups II-IV fibroblasts yield corresponding modest increases in PGE2 synthesis whose effects are progressively amplified through robust stabilization of COX-2 mRNA. PMID- 12543812 TI - Mechanisms by which tumor cells and monocytes expressing the angiogenic factor thymidine phosphorylase mediate human endothelial cell migration. AB - The angiogenic factor thymidine phosphorylase (TP) is highly expressed in many human solid tumors, and the level of its expression is associated with tumor neovascularization, invasiveness, and metastasis and with shorter patient survival time. TP promotes endothelial cell (EC) migration in vitro and angiogenesis in vivo, and these have been linked to its enzymatic activity. The mechanism by which TP stimulates EC migration was investigated using human umbilical vein ECs (HUVECs). TP induced concentration-dependent HUVEC migration, which required a TP gradient and thymidine and which was abrogated by the TP inhibitor CIMU (5-chloro-6(1-imidazolylmethyl)uracil). The chemotactic actions of TP plus thymidine were duplicated by the TP metabolite, 2-deoxyribose-1-phosphate (dR-1-P), and 10-fold more potently by its subsequent metabolite, 2-deoxyribose (2dR). Migration induced by dR-1-P, but not 2dR, was blocked by an alkaline phosphatase inhibitor, suggesting that the actions of dR-1-P first required its conversion to 2dR. In the migration assay, [5'-3H]dThd was metabolized to dR-1-P (96%) and 2dR (3.8%), and a gradient of both metabolites was maintained between the lower and upper chambers over the entire 5-h assay. TP expression in human solid tumors occurs in both tumor epithelial cells and in tumor-associated macrophages. The migration assay was adapted to use TP-transfected carcinoma cells to stimulate HUVEC migration, and they were found to induce more migration than did control vector-transfected cells. Human monocyte cells U937 and THP1, which constitutively expressed high levels of TP, also strongly induced HUVEC migration in the coculture assay. CIMU inhibited tumor-cell and monocyte-induced migration. In contrast, a neutralizing antibody to TP had no effect on cell stimulated HUVEC migration, even though it completely blocked the migration mediated by purified TP. Thus, the intracellular actions of TP were sufficient to stimulate HUVEC chemotaxis. In contrast to purified TP, when incubated with [5' 3H]-thymidine, cells expressing TP released up to 20-fold more 2dR into the medium than dR-1-P. These studies demonstrate that TP-expressing cells mediate EC migration via the intracellular metabolism of thymidine and subsequent extracellular release of 2dR, which forms a chemotactic gradient. PMID- 12543814 TI - Bcl-2 and Bax modulate adenine nucleotide translocase activity. AB - Bcl-2 is a prosurvival factor that reportedly prevents the nonspecific permeabilization of mitochondrial membranes, yet enhances specific ADP/ATP exchange by these organelles. Here, we show that Bcl-2 enhances the ADP/ATP exchange in proteoliposomes containing the purified adenine nucleotide translocase (ANT) in isolated mitochondria and mitoplasts, as well as in intact cells in which mitochondrial matrix ATP was monitored continuously using a specific luciferase-based assay system. Conversely, Bax, which displaces Bcl-2 from ANT in apoptotic cells, inhibits ADP/ATP exchange through a direct action on ANT. The Bax-mediated inhibition of ADP/ATP exchange can be separated from Bax stimulated formation of nonspecific pores by ANT. Chemotherapy-induced apoptosis caused an inhibition of ANT activity, which preceded the loss of the mitochondrial transmembrane potential and could be prevented by overexpression of Bcl-2. These data are compatible with a model of mitochondrial apoptosis regulation in which ANT interacts with either Bax or Bcl-2, which both influence ANT function in opposing manners. Bcl-2 would maintain the translocase activity at high levels, whereas Bax would inhibit the translocase function of ANT. PMID- 12543813 TI - Anti-CD20 therapeutic antibody rituximab modifies the functional organization of rafts/microdomains of B lymphoma cells. AB - Incubation of Burkitt lymphoma-derived Raji cells at physiological temperature with submicromolar concentrations of humanized anti-CD20 antibody rituximab (RTX) redistributes CD20 to liquid-ordered, plasma membrane rafts. This accumulation of the CD20 tetraspan protein in rafts does not change the existing lipid and phosphoprotein composition but makes sphingolipids and the Src regulator Cbp/PAG (Csk-binding protein/phosphoprotein associated with glycosphingolipid-enriched microdomain) transmembrane phosphoprotein more resistant to n-octyl-beta pyranoside, a detergent that dissociates sphingolipid clusters. On the contrary, sphingolipids and Cbp/PAG are not protected by the presence of CD20 against the disruptive effects of methyl-beta-cyclodextrin, a cyclic carbohydrate that removes membrane cholesterol. After accumulation of CD20, the activity of the raft-associated Lyn kinase is down-regulated without apparent alteration of its relationship to substrates. Moreover, in rafts of lymphoblastoid cells that express lower amounts of Cbp/PAG, RTX redistributes CD20 to rafts but does not modulate the raft-associated protein tyrosine kinase activity, suggesting that the presence of Cbp/PAG protein in rafts is necessary for RTX to exert its transmembrane "signaling effects." Lastly, redistribution of CD20 in rafts renders the glycosylphosphatidyl inositol (GPI)-linked CD55 C'-defense protein hypersensitive to glycosylphosphatidyl inositol-specific phospholipases. By redistributing CD20 to rafts, RTX modifies their stability and organization and modulates the associated signaling pathways and C' defense capacity. PMID- 12543815 TI - The causes of autism spectrum disorders. PMID- 12543816 TI - Governance for NHS foundation trusts. PMID- 12543817 TI - Reinstitutionalisation in mental health care. PMID- 12543818 TI - BMJ Learning. PMID- 12543819 TI - Non-invasive ventilation in chronic obstructive pulmonary disease. PMID- 12543820 TI - Health professionals challenge US smallpox vaccination plan. PMID- 12543823 TI - Police uncover large scale organ trafficking in Punjab. PMID- 12543826 TI - The market for medical journals is anticompetitive, says expert. PMID- 12543825 TI - FDA halts gene therapy trials after leukaemia case in France. PMID- 12543829 TI - US health care should focus on 20 areas to increase impact. PMID- 12543830 TI - Restoring the profession's self esteem. PMID- 12543831 TI - Taking up cudgels for peace. PMID- 12543832 TI - Non-invasive positive pressure ventilation to treat respiratory failure resulting from exacerbations of chronic obstructive pulmonary disease: Cochrane systematic review and meta-analysis. AB - OBJECTIVES: To determine the effectiveness of non-invasive positive pressure ventilation (NPPV) in the management of respiratory failure secondary to acute exacerbation of chronic obstructive pulmonary disease. DESIGN: Systematic review of randomised controlled trials that compared NPPV and usual medical care with usual medical care alone in patients admitted to hospital with respiratory failure resulting from an exacerbation of chronic obstructive pulmonary disease and with PaCO2 >6 kPa. RESULTS: The eight studies included in the review showed that, compared with usual care alone, NPPV as an adjunct to usual care was associated with a lower mortality (relative risk 0.41 (95% confidence interval 0.26 to 0.64)), a lower need for intubation (relative risk 0.42 (0.31 to 0.59)), lower likelihood of treatment failure (relative risk 0.51 (0.38 to 0.67)), and greater improvements at 1 hour in pH (weighted mean difference 0.03 (0.02 to 0.04)), PaCO2 (weighted mean difference -0.40 kPa (-0.78 to -0.03)), and respiratory rate (weighted mean difference -3.08 breaths per minute (-4.26 to 1.89)). NPPV resulted in fewer complications associated with treatment (relative risk 0.32 (0.18 to 0.56)) and shorter duration of stay in hospital (weighted mean difference -3.24 days (-4.42 to -2.06)). CONCLUSIONS: NPPV should be the first line intervention in addition to usual medical care to manage respiratory failure secondary to an acute exacerbation of chronic obstructive pulmonary disease in all suitable patients. NPPV should be tried early in the course of respiratory failure and before severe acidosis, to reduce mortality, avoid endotracheal intubation, and decrease treatment failure. PMID- 12543833 TI - NHS waiting lists and evidence of national or local failure: analysis of health service data. AB - OBJECTIVES: To investigate the national distribution of prolonged waiting for elective day case and inpatient surgery, and to examine associations of prolonged waiting with markers of NHS capacity, activity in the independent sector, and need. SETTING: NHS hospital trusts in England. POPULATION: People waiting for elective treatment in the specialties of general surgery; ear, nose and throat surgery; ophthalmic surgery; and trauma and orthopaedic surgery. MAIN OUTCOME MEASURE: Numbers of people waiting six months or longer (prolonged waiting). Characteristics of trusts with large numbers waiting six months or longer were examined by using logistic regression. RESULTS: The distribution of numbers of people waiting for day case or elective surgery in all the specialties examined was highly positively skewed. Between 52% and 83% of patients waiting longer than six months in the specialties studied were found in one quarter of trusts, which in turn contributed 23-45% of the national throughput specific to the specialty. In general, there was little evidence to show that capacity (measured by numbers of operating theatres, dedicated day case theatres, available beds, and bed occupancy rate) or independent sector activity were associated with prolonged waiting, although exceptions were noted for individual specialties. There was consistent evidence showing an increase in prolonged waiting, with increased numbers of anaesthetists across all specialties and with increased bed occupancy rates for ear, nose and throat surgery. Markers of greater need for health care, such as deprivation score and rate of limiting long term illness, were inversely associated with prolonged waiting. CONCLUSION: In most instances, substantial numbers of patients waiting unacceptably long periods for elective surgery were limited to a small number of hospitals. Little and inconsistent support was found for associations of prolonged waiting with markers of capacity, independent sector activity, or need in the surgical specialties examined. PMID- 12543834 TI - Case fatality rates after admission to hospital with stroke: linked database study. PMID- 12543835 TI - Career choices of United Kingdom medical graduates of 1999 and 2000: questionnaire surveys. PMID- 12543836 TI - Barriers to accurate diagnosis and effective management of heart failure in primary care: qualitative study. AB - OBJECTIVE: To ascertain the beliefs, current practices, and decision making of general practitioners in the diagnosis and management of suspected heart failure in primary care, with a view to identifying barriers to good care. DESIGN: A qualitative approach using focus groups with 30 general practitioners from four primary care groups. The sampling strategy was stratified and purposive. The contents of interviews were transcribed and analysed according to the principles of "pragmatic variant" grounded theory. SETTING: North east England. RESULTS: Three categories of difficulties contribute to variations in medical practice and to the reasons why general practitioners experience difficulties in diagnosing and managing heart failure. The first is uncertainty about clinical practice, including lack of confidence in establishing an accurate diagnosis and worries about using angiotensin converting enzyme inhibitors, beta blockers, and spironolactone in patients who are often elderly and frail, with comorbidity and polypharmacy. The second is a lack of awareness of relevant research evidence in what was perceived to be a complex and rapidly changing therapeutic field. Doubts about the applicability of research findings in primary care, and fear of information overload also emerged. The third category consists of influences of individual preference and local organisational factors. Medical training, negative clinical experiences, and outside agencies influenced the behaviour of general practitioners and professional culture. Local factors included the availability of diagnostic services, resources (such as accessible cardiologists), and interactions between professionals in primary or secondary care, and they seemed to shape the practice and decision making processes in primary care. CONCLUSIONS: The national service framework for coronary heart disease stresses that the substandard care of patients with heart failure is unacceptable. This study identified barriers to be overcome across primary and secondary care in implementation strategies that are specific to the locality and multifaceted. Single strategies--for example, the provision of guidelines--are unlikely to have an impact on clinical outcomes, and new, conjoint models of care need to be explored. PMID- 12543837 TI - Identifying depression in primary care: a comparison of different methods in a prospective cohort study. PMID- 12543838 TI - Ten ways to improve information technology in the NHS. PMID- 12543839 TI - Onchocerciasis. PMID- 12543840 TI - Fear of the dark in children: is stationary night blindness the cause? PMID- 12543841 TI - Applying educational theory in practice. PMID- 12543842 TI - WHO responds. PMID- 12543843 TI - Interaction revisited: the difference between two estimates. PMID- 12543844 TI - Open letter to Tony Blair: call to prevent escalating violence. PMID- 12543845 TI - Doctors and computers. Poor system design and little investment mean hospital doctors do not use computers . . . PMID- 12543847 TI - Risk factors for cot death increase danger from infection. PMID- 12543848 TI - Long term effects of advice to reduce dietary salt. Front cover was highly misleading. PMID- 12543849 TI - Indian objection to export of human tissue for research. Clarifications from authors of study. PMID- 12543850 TI - Ethics dialogue between rich and poor countries is overdue. PMID- 12543851 TI - Issues relating to abortions are complicated in Nigeria. PMID- 12543852 TI - Orthopaedic surgery. PMID- 12543853 TI - Gaining experience in different teaching methods. PMID- 12543857 TI - Transgenic sickle mice have vascular inflammation. AB - Inflammation may play an essential role in vaso-occlusion in sickle cell disease. Sickle patients have high white counts and elevated levels of serum C-reactive protein (CRP), cytokines, and adhesion molecules. In addition, circulating endothelial cells, leukocytes, and platelets are activated. We examined 4 transgenic mouse models expressing human alpha- and sickle beta-globin genes to determine if they mimic the inflammatory response seen in patients. These mouse models are designated NY-S, Berk-S(Antilles), NY-S/S(Antilles) (NY-S x Berk S(Antilles)), and Berk-S. The mean white counts were elevated 1.4- to 2.1-fold (P 70% DSP), was observed at 4000 s-1 but not at 200 s-1. Monoclonal antibodies (MoAbs) 6D1 (anti-GPIb) and 328 (anti-VWF A1 domain), completely abolished SIPA and p125FAK phosphorylation mediated by 2B-rVWF. In contrast, neither RGDS peptide nor MoAb 7E3, both known to block alphaIIbbeta3 engagement, had any effect on SIPA and pp125FAK. The size of aggregates formed at a high shear rate in the presence of 2B-rVWF was decreased by genistein, demonstrating the biologic relevance of pp125FAK. These findings provide a unique mechanism whereby the enhanced interaction of 2B-rVWF with GPIb, without engagement of alphaIIbbeta3, is sufficient to induce SIPA but does not lead to stable thrombus formation. PMID- 12543871 TI - Nodular sclerosing Hodgkin disease: new grading predicts prognosis in intermediate and advanced stages. AB - The prognostic value of histologic classification and single histomorphologic parameters in Hodgkin disease has been widely debated in the literature. Whereas several former studies identified single parameters to be of clinical relevance, some recent reports have doubted the prognostic value of histology using modern treatment. Grading of the largest histologic category of Hodgkin disease, nodular sclerosis (NS), has been controversially discussed concerning clinical relevance. In this study, 965 cases of NS were reviewed to assess 9 histomorphologic parameters. The histologic results were correlated with laboratory and clinical findings and with overall survival and disease-free survival. Based on these results, a new grading of the NS category was established. The new grading, based on the 3 criteria eosinophilia, lymphocyte depletion, and atypia of the Hodgkin/Reed-Sternberg cells, was a significant indicator of prognosis in intermediate and advanced stages. Patients investigated in this study represent an outstanding collection because all of them were enrolled in the prospective multicenter clinical trial of the German Hodgkin Lymphoma Study Group. All of them had been staged uniformly according to the Ann Arbor system and had received stage-adapted modern treatment according to multimodality protocols. A subtle analysis of histology could represent a possible way to identify patients with a significantly better or worse prognosis. This new grading should help to avoid overtreatment to reduce severe therapy-related side effects such as acute toxicity and chronic sequelae such as cardiopulmonary complications and secondary neoplasias. PMID- 12543872 TI - Public health, GIS, and the internet. AB - Internet access and use of georeferenced public health information for GIS application will be an important and exciting development for the nation's Department of Health and Human Services and other health agencies in this new millennium. Technological progress toward public health geospatial data integration, analysis, and visualization of space-time events using the Web portends eventual robust use of GIS by public health and other sectors of the economy. Increasing Web resources from distributed spatial data portals and global geospatial libraries, and a growing suite of Web integration tools, will provide new opportunities to advance disease surveillance, control, and prevention, and insure public access and community empowerment in public health decision making. Emerging supercomputing, data mining, compression, and transmission technologies will play increasingly critical roles in national emergency, catastrophic planning and response, and risk management. Web-enabled public health GIS will be guided by Federal Geographic Data Committee spatial metadata, OpenGIS Web interoperability, and GML/XML geospatial Web content standards. Public health will become a responsive and integral part of the National Spatial Data Infrastructure. PMID- 12543873 TI - Complement inhibition in renal diseases. PMID- 12543874 TI - Evidence-based recommendations for immunosuppression in IgA nephropathy: handle with caution. PMID- 12543875 TI - Recent approaches to the pathogenesis of minimal-change nephrotic syndrome. PMID- 12543876 TI - Arterial pressure components and cardiovascular risk in end-stage renal disease. PMID- 12543877 TI - New insights in dialysis membrane biocompatibility: relevance of adsorption properties and heparin binding. PMID- 12543878 TI - Creatine supplementation does not affect kidney function in an animal model with pre-existing renal failure. AB - BACKGROUND: Creatine is widely used as an ergogenic substance among athletes. Safety of prolonged creatine intake has been questioned, based upon case reports and animal data. We investigated the effect of prolonged creatine ingestion on renal function in animals with normal kidney function or pre-existing kidney failure, respectively. METHODS: Male Wistar rats were randomly allocated to four experimental groups: (i) sham-operated, control diet; (ii) sham-operated, creatine-supplemented diet (2% w/w (0.9+/-0.2 g creatine/kg body weight/day)); (iii) two-thirds nephrectomized, control diet; and (iv) two-thirds nephrectomized, creatine supplemented diet. Glomerular filtration rate was determined using inulin and creatinine clearance, together with albumin excretion, urea clearance, muscle and serum creatine and serum cystatin C concentrations. RESULTS: In contrast to previous reports, no detrimental effects of creatine supplementation on the renal function indices were observed in two thirds nephrectomized or sham-operated animals. No differences were observed in inulin (0.28+/-0.08 vs 0.25+/-0.08 ml/min/100 g; P=NS) or creatinine clearance rates. Serum cystatin C concentration, urinary protein excretion, and albumin and urea clearance were comparable between creatine-supplemented and control-diet fed animals in both sham-operated and two-thirds nephrectomized animals. Serum creatine and intramuscular total creatine concentrations were higher in creatine supplemented groups (P<0.05). CONCLUSIONS: Creatine supplementation at a dosage of 2% w/w for 4 weeks does not impair kidney function in animals with pre existing renal failure or in control animals. PMID- 12543879 TI - HMG-CoA reductase inhibitor ameliorates diabetic nephropathy by its pleiotropic effects in rats. AB - BACKGROUND: An inflammatory process may be one of the critical factors that contribute to the development of diabetic nephropathy (DN). We reported previously that intercellular adhesion molecule-1 (ICAM-1) is up-regulated and promotes macrophage infiltration in the glomeruli of diabetic rats. 3-Hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) have recently been emphasized to have anti-inflammatory effects; inhibition of leukocyte adhesion and migration, independent of the cholesterol-lowering effect. The present study was designed to test the hypothesis that statins prevent the development of DN by pleiotropic effects. METHODS: Streptozotocin-induced diabetic rats were treated with cerivastatin (0.5 mg/kg body weight) or vehicle for 4 weeks. We analysed glomerular macrophage infiltration and ICAM-1 expression. We also evaluated major regulators of ICAM-1, activation of nuclear factor-kappa B (NF-kappaB) using electrophoretic mobility shift assay, and oxidative stress. RESULTS: Statin treatment reduced urinary albumin excretion (UAE) (2.96+/-0.18 vs 2.38+/-0.06; log(10) UAE, P<0.05), glomerular size (12 150+/-329 vs 9963+/-307 micro m(2), P<0.05), and lowered blood pressure, compared with untreated diabetic rats. Immunohistochemistry revealed that macrophage infiltration and ICAM-1 expression in glomeruli were increased in diabetic rats and were inhibited by statin treatment. Renal NF-kappaB activity, urinary excretion and renal deposition of 8-OHdG were increased in diabetic rats, and reduced by statin treatment. CONCLUSION: Statin treatment prevented glomerular injury, independent of the cholesterol-lowering effects. Our findings suggest that the beneficial effect might be mediated by pleiotropic effects including an anti-inflammatory action through a reduction of oxidative stress, NF-kappaB activation, ICAM-1 expression and macrophage infiltration in the early phase of DN. PMID- 12543880 TI - Primary hyperoxaluria type 1 in The Netherlands: prevalence and outcome. AB - BACKGROUND: Primary hyperoxaluria type 1 (PH1) is a phenotypically heterogeneous disease. To date the relationship between biochemical parameters and outcome is unclear. We therefore undertook a national cohort study on biochemical and clinical parameters and outcome in PH1. METHODS: Review of medical charts of all Dutch PH1 patients, who were identified by sending questionnaires to all Dutch nephrologists for children and adults. RESULTS: Fifty-seven patients were identified. The prevalence and incidence rates were 2.9/10(6) and 0.15/10(6)/year, respectively. Median age at diagnosis was 7.3 years (range 0 57). Seventeen (30%) patients were older than 18 years at time of diagnosis, of whom 10 (59%) presented with end-stage renal disease (ESRD), in contrast to only nine (23%) of those aged under 18 years. Median age at initial symptoms was 6.0 years (range 0-50). In four of nine patients with infantile PH1, normal renal function was preserved after a median follow-up of 7.7 years (range 0.1-16). Progression to renal insufficiency was associated with the presence of nephrocalcinosis, as assessed by ultrasound (relative risk=1.8; 95% CI, 1.0-3.4) and with pyridoxine-unresponsiveness (relative risk=2.2; 95% CI, 1.1-4.2) but not with age at presentation, the extent of hyperoxaluria, or AGT activity. No apparent nephrocalcinosis was found in five of the 19 patients who presented with ESRD. CONCLUSIONS: Although more than one-half of the PH1 patients have symptoms under the age of 10 years, PH1 can present at any age. In adults, PH1 presents predominantly with ESRD, which may be due to misinterpretation of early symptoms. Although nephrocalcinosis is correlated with development of renal insufficiency, the latter can occur even in the absence of nephrocalcinosis. Pyridoxine sensitivity is associated with better outcome in PH1. PMID- 12543881 TI - Seven cases of granulomatous interstitial nephritis in the absence of extrarenal sarcoid. AB - BACKGROUND: Renal disease in sarcoidosis may occur due to granulomatous interstitial nephritis. However, granulomatous interstitial nephritis in the absence of features of extrarenal sarcoid, or other causes, has been reported very rarely. In this report we describe seven such patients. METHODS: Since 1995, we have identified a number of patients with biopsy-proven granulomatous interstitial nephritis. Patients were excluded if they had (i) evidence of extrarenal sarcoid, (ii) infections that may have contributed to pathogenesis or (iii) an obvious drug-related aetiology. RESULTS: Seven patients were identified, of whom five were male and two female, with a median age of 69. Median calculated creatinine clearance at presentation was 14 ml/min. Two had raised serum calcium at presentation and three had a raised serum angiotensin-converting enzyme. All patients were treated with steroids and five out of seven had an improvement in their renal function. Two patients progressed to end-stage renal failure despite treatment with steroids. CONCLUSIONS: Idiopathic granulomatous interstitial nephritis may represent a renal-limited form of sarcoid. It may be associated with hypercalcaemia and a raised serum angiotensin-converting enzyme and usually responds to treatment with corticosteroids. PMID- 12543882 TI - Upregulation of MHC class II, interferon-alpha and interferon-gamma receptor protein expression in HIV-associated nephropathy. AB - BACKGROUND: Renal cellular HIV infection has been linked to the pathogenesis of HIV-associated nephropathy (HIVAN), but mediators of its development are unknown. HIV infection is associated with disordered cytokine metabolism, and chemokine receptors are coreceptors for HIV immune cellular infection. Chemokines such as interleukin (IL)-8, monocyte chemoattractant protein-1 (MCP-1) and RANTES, and interferons (IFNs) have been implicated in the progression of nephropathy. Renal major histocompatibility complex (MHC) protein expression is involved in antigen presentation and modulating tissue cellular immune responses. Their relative importance in HIVAN pathogenesis is unknown. METHODS: We measured levels of chemokines, IFN-alpha, IFN-gamma receptor and non-polymorphic MHC Class II protein by high performance capillary electrophoresis, and incubation with antibodies for quantification by chemiluminesce in renal tissue of patients with HIVAN, compared with tissue without HIV infection, in the presence and absence of nephropathy. Renal biopsy tissue protein levels were correlated with the number and type of infiltrating tissue immune cells. RESULTS: Mean renal interstitial and glomerular MCP-1, RANTES and IL-8 tissue levels were higher in patients with HIV infection compared with tissue without HIV infection, regardless of the presence of renal disease. In contrast, mean renal interstitial and glomerular non-polymorphic MHC Class II, IFN-alpha and IFN-gamma receptor protein were higher in patients with HIVAN compared with all other groups. Tissue MHC Class II and IFN-gamma receptor protein levels did not correlate with immune cellular infiltration in patients with HIV infection and renal disease. CONCLUSIONS: The data suggest an upregulated renal immune microenvironment, capable of antigen presentation, exists in HIVAN. MHC Class II proteins and IFNs, and the capacity to present antigen may be crucial in HIVAN pathogenesis. PMID- 12543883 TI - Time course of the antiproteinuric and antihypertensive effect of losartan in diabetic nephropathy. AB - BACKGROUND: Blockade of the renin-angiotensin system is the primary target in the treatment of diabetic kidney disease. Angiotensin II subtype 1 (AT1) receptor antagonists reduce albuminuria and lower blood pressure, but the initial time course of these effects after initiation of treatment is unknown. We evaluated the time course of the antihypertensive and antialbuminuric effect after initiation of AT1 receptor blockade by losartan in diabetic nephropathy. METHODS: Ten hypertensive type 1 diabetic patients with diabetic nephropathy were included in the study. After a washout period of 4 weeks, patients received losartan 100 mg once daily for 28 days. Every morning, one urine sample was collected for daily determination of albumin/creatinine ratio. Twenty-four hour blood pressure (Takeda TM2420), plasma renin and plasma creatinine were measured at baseline and days 7, 14 and 28. RESULTS: Baseline levels of urinary albumin/creatinine ratio and 24 h mean arterial blood pressure were 676 (402-1136) mg/g (geometric mean and 95% CI, respectively) and 100+/-3 mmHg (mean +/- SEM). Albumin/creatinine ratio was significantly reduced after 7 days of treatment by 29% (15-41) (95% CI) without significant further reductions during the 28 day study period (P<0.01 vs baseline). Mean arterial blood pressure was significantly lowered by 7 mmHg after 7 days of treatment and remained unchanged throughout the study (P<0.01 vs baseline). Plasma renin was significantly increased from baseline after initiation of losartan treatment and stabilized after 7 days (P<0.01). We found no changes in plasma creatinine during the study. CONCLUSIONS: The initial time course of the reduction in arterial blood pressure and albuminuria are concordant, which suggests that systemic and renal haemodynamic mechanisms are of primary importance in the reduction of albuminuria. PMID- 12543884 TI - Continuing uncertainty about the value of percutaneous revascularization in atherosclerotic renovascular disease: a meta-analysis of randomized trials. AB - BACKGROUND: To study the effect of revascularization on blood pressure (BP) and serum creatinine (SCr) in patients with atherosclerotic renovascular disease (ARVD). METHODS: Three randomized studies comparing balloon angioplasty (plus medication if necessary) with medical therapy alone in patients with ARVD were identified. In one study, patients were stratified and analysed according to whether they had unilateral or bilateral disease. Therefore, four sets of results were available for inclusion in a meta-analysis comparing BP and SCr at 6 months and changes from baseline. RESULTS: The three trials recruited 210 patients. There was no clear benefit for angioplasty when comparing BP at 6 months. Relative to the medical therapy group, the mean (95% CI) systolic/diastolic BP was 2.9 mmHg (-9.1, 3.4)/0.35 mmHg (-3.6, 2.9) lower in the angioplasty group (P=0.4/0.8). There was, however, some suggestion of benefit for angioplasty when changes in BP were compared. There was a greater reduction in the systolic/diastolic BP in the angioplasty group, with a difference of 6.3 mmHg ( 11.7, -0.8)/3.3 mmHg (-6.2, -0.4) in the mean change (P=0.02/0.03). There was some suggestion of benefit for angioplasty in terms of changes in SCr, although this was not significant (P=0.06). CONCLUSIONS: The reported trials have been too small to determine reliably the role of angioplasty in ARVD. Although the combined results of three previous trials exclude the possibility of a large improvement in renal function or hypertension after angioplasty, a moderate but clinically worthwhile benefit cannot be ruled out. Further large-scale randomized evidence is needed. PMID- 12543885 TI - Attitudes of Canadian nephrologists, family physicians and patients with kidney failure toward primary care delivery for chronic dialysis patients. AB - BACKGROUND: Nephrologists have traditionally assumed responsibility for both nephrological and primary care health problems of their dialysis patients. However, given the increasing limitations of nephrology human resources, there is concern that traditional models may fall short of providing comprehensive care. METHODS: We studied this issue by distributing three different self-administered surveys to 361 members of the Canadian Society of Nephrology, 325 family physicians, and 163 chronic dialysis patients. RESULTS: The overall response rate was 61.3% for nephrologists, 51% for family physicians, and 90% for patients. More than 50% of Canadian nephrologists are spending approximately one-third of their time in primary care delivery. The majority of these nephrologists and family physicians agree that nephrologists should not be solely responsible for the primary care of patients on dialysis. Yet, both groups of physicians have concerns that family physicians do not have the knowledge/training and time to care for this complicated group of patients. The patients themselves have more confidence in the primary care that is delivered by their family physicians than by their nephrologists. Unfortunately, there is little communication between the two physician groups either between themselves or with their patients about the services that should be provided by their nephrologist or their family physician. CONCLUSION: Nephrologists and family physicians agree that more primary care for dialysis patients should be provided by family physicians. However, the lack of communication between physicians and patients may result in either a duplication or omission of services that are required by this patient population. Dialysis delivery systems in Canada must evolve to ensure that comprehensive chronic dialysis and primary care is provided to these patients through cooperation and communication with primary care physicians. PMID- 12543886 TI - Quality of life in adults with end-stage renal disease since childhood is only partially impaired. AB - BACKGROUND: Little is known about the quality of life of adults with end-stage renal disease (ESRD) since childhood. In a long-term follow-up study, we assessed quality of life in these patients and compared their outcomes with those in the general population and in dialysis patients with adult-onset of ESRD. METHODS: All Dutch adult patients with onset of ESRD at age 0-14 years between 1972 and 1992 were asked to complete the RAND-36 questionnaire. We obtained clinical information by reviewing all available medical charts, and by current history. Patient scores were compared with those of age-matched healthy controls and with those of patients with adult onset of dialysis, of whom data were derived from a national study on the adequacy of dialysis (NECOSAD-2). RESULTS: We obtained a complete RAND-36 score in 131 of 187 surviving patients. Clinical characteristics of participants and non-participants were similar. Compared with the general population, we found a higher prevalence of impaired quality of life in dialysis patients in the following domains: physical functioning, role limitations due to physical health, social functioning (SF), general health perception (GH) and physical component summary. In other domains, impaired quality of life was equally or less prevalent (mental component summary). In transplanted patients, only impaired GH and SF were more prevalent. In dialysis patients with adult ESRD onset, impaired quality of life was more prevalent than in the general population in all domains, except in emotional role limitations. CONCLUSIONS: Dialysis patients with paediatric ESRD have an impaired physical quality of life, but in contrast to dialysis patients with adult ESRD onset, they have a normal mental quality of life. PMID- 12543887 TI - Prognostic value of heart rate variability in patients with end-stage renal disease on chronic haemodialysis. AB - BACKGROUND: Although decreased heart rate variability (HRV) is an independent predictor of death in various populations, its prognostic value in patients with end-stage renal disease on chronic haemodialysis is unknown. METHODS: We prospectively studied 120 chronic haemodialysis patients (age 61+/-11 years; males 51%; diabetics 38%; duration of haemodialysis therapy 50+/-114 months) who underwent 24 h electrocardiography at baseline for analysis of time- and frequency-domain HRV. RESULTS: All HRV measures in the patients were significantly reduced compared with those obtained from 62 age-matched healthy subjects. During a follow-up period of 26+/-10 months, 21 patients died (17.5%); 10 from cardiac causes and 11 from non-cardiac causes (seven fatal strokes and four other causes). A Cox proportional hazards model revealed that, of the HRV measures, decreases in the triangular index (TI), very-low-frequency (0.0033-0.04 Hz) power, ultra-low-frequency (<0.0033 Hz) power (ULF) and the ratio of low frequency (0.04-0.15 Hz) power to high-frequency (0.15-0.4 Hz) power had significant predictive value for cardiac death. None of the HRV measures, however, had predictive value for non-cardiac death, including stroke death. Even after adjustment for other univariate predictors including age, diabetes, serum albumin and coronary artery disease, the predictive value of decreased TI and ULF remained significant-adjusted relative risk (95% confidence interval) per 1 SD decrement of TI and ULF, 3.28 (1.08-9.95) and 1.92 (1.01-3.67), respectively. CONCLUSIONS: Decreases in some HRV measures, particularly those reflecting long term variability, are independent predictors of cardiac death in chronic haemodialysis patients. PMID- 12543888 TI - Anthropometric measures, cytokines and survival in haemodialysis patients. AB - BACKGROUND: Lower serum albumin concentration (sAlb) and higher levels of pro inflammatory cytokines have been reported to predict death in patients treated with haemodialysis (HD). SAlb, along with anthropometric measures, has been used as a surrogate marker for nutritional status in patients with chronic disease. Though adequate nutrition has been considered an important factor for patients treated with HD, it has not been established if any nutritional markers other than lower serum albumin and lower body mass index (BMI) predict death. Furthermore, it has not been shown whether anthropometric measures other than BMI are associated with predictors of mortality. METHODS: At the outset of the study, patients were assessed using demographic and anthropometric indices including arm fat area (AFA), arm muscle area (AMA), BMI, per cent ideal weight (PIW), pre dialysis sAlb, and circulating levels of tumour necrosis factor-alpha (TNF alpha), IL-1 and IL-6. A severity index, previously demonstrated to be a mortality marker, was used to grade medical co-morbidity. RESULTS: Two-hundred and forty patients entered the study. The mean age was 55.1+/-14.3 years, mean sAlb 3.76+/-0.60 mg/dl, mean AFA 1742+/-1225 mm(2), mean AMA 5464+/-1817 mm(2), mean PIW 101.0+/-21.3% and mean BMI 24.9+/-5.6 kg/m(2). PIW, BMI, AFA and AMA were, as expected, all highly correlated with one another. SAlb correlated with serum transferrin; however, neither sAlb nor serum transferrin concentration correlated with circulating cytokine levels. Circulating cytokines and sAlb did not correlate with PIW, BMI, AFA or AMA. In Cox regression analyses using multiple control variables, IL-6 predicted survival, while the anthropometric measures did not. CONCLUSIONS: Pro-inflammatory cytokines and sAlb are robust predictors of death in patients treated with HD. PIW and BMI correlate well with other anthropometric measures in patients treated with HD, but these measures do not correlate with markers of inflammation. Anthropometric measures are poor predictors of survival compared with measures linked to the acute-phase response. PMID- 12543889 TI - Influence of body composition on 5 year mortality in patients on regular haemodialysis. AB - BACKGROUND: Reduction of body mass index (BMI) significantly affects mortality in haemodialysis (HD) patients, but it remains to be determined which of the body components influences mortality. METHODS: We examined the whole body composition of 262 HD patients by dual-energy X-ray absorptiometry (DEXA) (age: 60+/-12 years; HD duration 9+/-7 years; male/female: 177/85; diabetics, n=50) and subsequently followed mortality for 5 years. RESULTS: Patient age was significantly correlated with limb/trunk lean mass (LTLM) ratio (r=-0.350, P<0.01) and % fat content in whole tissue (r=0.145, P=0.02). There was a significant positive relationship between LTLM ratio and serum creatinine both in males (r=0.404, P<0.01) and females (r=0.267, P=0.01). Diabetic males and females both had a significantly lower LTLM ratio than non-diabetic males (P<0.01) and females (P<0.04). During the 5 years, 65 patients (24.8%) died mainly of cardiovascular diseases and infections. BMI was lower in the expired group than in survivors (P<0.04). LTLM ratio was significantly reduced in the expired group compared with the surviving males (0.629+/-0.097 vs 0.707+/-0.094; P<0.01) and females (0.611+/-0.101 vs 0.651+/-0.078; P<0.01). Cox's proportional hazards analysis revealed that the reduction of LTLM ratio was a significant determinant of death in men (P<0.01), while a lower percentage of fat content of trunk was a significant determinant of death in women (P<0.01). In contrast, BMI did not influence mortality in either sex. CONCLUSIONS: Measurements of regional lean and fat mass volumes by DEXA may be useful for predicting death in patients receiving long-term HD. PMID- 12543890 TI - Dietary potassium and laxatives as regulators of colonic potassium secretion in end-stage renal disease. AB - BACKGROUND: In end-stage renal disease (ESRD), colonic potassium (K+) secretion increases as renal K+ excretion declines. The nature of this adaptive process is poorly understood, but post-prandial increases in plasma K+ concentration may be a determining factor. In addition, even though colonic K+ secretion increases in ESRD, interdialytic hyperkalaemia is a serious problem in haemodialysis patients, which might be reduced by stimulating colonic K+ secretion still further using laxatives. METHODS: Plasma K+ concentrations were measured in the fasting state, and for 180 min after the oral administration of 30 mmol of K+ to nine control subjects and 16 normokalaemic patients with ESRD (eight "predialysis" patients and eight patients undergoing continuous ambulatory peritoneal dialysis (CAPD)). Plasma K+ concentrations were also monitored for 180 min in fasting controls and ESRD patients who were not given the oral K+ load. To study the effect of laxatives on interdialytic hyperkalaemia, plasma K+ concentrations were measured in eight control subjects and 13 haemodialysis patients before and during 2 weeks treatment with bisacodyl (a cAMP-mediated laxative) and in five haemodialysis patients before and during 2 weeks treatment with lactulose (an osmotic laxative). RESULTS: Oral K+ loading caused plasma K+ concentration to rise within the normal range (3.5-5.1 mmol/l) in control subjects, while significantly higher concentrations were achieved in the "predialysis" patients and sustained hyperkalaemia developed in the CAPD patients. Bisacodyl treatment had no effect on plasma K+ concentrations in control subjects, but significantly decreased the mean interdialytic plasma K+ concentration (from 5.9+/-0.2 to 5.5+/-0.2 mmol/l, P<0.0005) in haemodialysis patients, whereas plasma K+ concentration did not change during lactulose treatment. CONCLUSIONS: Higher plasma K+ concentrations after food may help to maintain K+ homeostasis in ESRD by enhancing colonic K+ secretion. Bisacodyl may be useful for reducing interdialytic hyperkalaemia in patients undergoing haemodialysis. PMID- 12543891 TI - Clinical significance of SEN virus infection in patients on maintenance haemodialysis. AB - BACKGROUND: The SEN virus (SENV) has been identified as a putative new hepatitis virus. This study was conducted to clarify the clinical significance of SENV infection in patients on haemodialysis. METHODS: A total of 189 patients on maintenance haemodialysis and 60 healthy controls were enrolled. Of the 189 patients, 154 were followed up for 2 years. SENV DNA (genotypes D and H) was measured by means of polymerase chain reaction. RESULTS: SENV infection was significantly (P=0.012) more prevalent in patients on haemodialysis (38%) than in controls (22%). SENV infection was not associated with the amount of transfusion or duration of haemodialysis, while hepatitis C virus (HCV) infection was significantly associated with both of these factors. Elevation of alanine aminotransferase was significantly associated with HCV, but not with SENV viraemia. Of the 154 patients who were followed up, 63 (41%) remained negative, 34 (22%) gained positivity, 28 (18%) lost it and 29 (19%) remained positive for SENV infection. Episodes of alanine aminotransferase elevation were recorded for 3% of the patients who incurred SENV infection and this rate was similar to that observed in patients who were continuously negative for SENV infection (5%). CONCLUSIONS: SENV infection was common in patients on haemodialysis. No evidence was obtained that suggested involvement of the hepatitis virus in the pathogenicity of SENV. PMID- 12543892 TI - A randomized controlled trial of haemoglobin normalization with epoetin alfa in pre-dialysis and dialysis patients. AB - BACKGROUND: Partial correction of renal anaemia with erythropoietin improves quality of life (QoL). We aimed to examine if normalization of haemoglobin with epoetin alfa in pre-dialysis and dialysis patients further improves QoL and is safe. METHODS: 416 Scandinavian patients with renal anaemia [pre-dialysis, haemodialysis (HD) and peritoneal dialysis patients] were randomized to reach a normal haemoglobin of 135-160 g/l (n=216) or a subnormal haemoglobin of 90-120 g/l (n=200) with or without epoetin alfa. Study duration was 48-76 weeks. QoL was measured using Kidney Disease Questionnaires in 253 Swedish dialysis patients. Safety was examined in all patients. RESULTS: QoL improved, measured as a decrease in physical symptoms (P=0.02), fatigue (P=0.05), depression (P=0.01) and frustration (P=0.05) in the Swedish dialysis patients when haemoglobin was normalized. In pre-dialysis patients, diastolic blood pressure was higher in the normal compared with the subnormal haemoglobin group after 48 weeks. However, the progression rate of chronic renal failure was comparable. In the normal haemoglobin group (N-Hb), 51% had at least one serious adverse event compared with 49% in the subnormal haemoglobin group (S-Hb) (P=0.32). The incidence of thrombovascular events and vascular access thrombosis in HD patients did not differ. The mortality rate was 13.4% in the N-Hb group and 13.5% in the S-Hb group (P=0.98). Mortality decreased with increasing mean haemoglobin in both groups. CONCLUSIONS: Normalization of haemoglobin improved QoL in the subgroup of dialysis patients, appears to be safe and can be considered in many patients with end-stage renal disease. PMID- 12543893 TI - Treatment of anaemia in dialysis patients with unit dosing of darbepoetin alfa at a reduced dose frequency relative to recombinant human erythropoietin (rHuEpo). AB - BACKGROUND: Darbepoetin alfa is an erythropoietic agent with a 3-fold longer elimination half-life than recombinant human erythropoietin (rHuEpo), which allows less frequent dosing. This study investigated the safety and efficacy of darbepoetin alfa for treating anaemia in dialysis patients, using a dosing regimen that was independent of the patient's body weight (unit dosing). METHODS: Dialysis patients (n=341) maintained on rHuEpo treatment (alfa or beta) were switched to darbepoetin alfa at a reduced dosing frequency, but by the same route of administration [intravenous (i.v.) or subcutaneous (s.c.)]. Patients receiving rHuEpo two or three times weekly changed to once-weekly darbepoetin alfa, and those receiving rHuEpo once weekly changed to once every other week darbepoetin alfa. The unit doses of darbepoetin alfa (10-150 microg) were titrated to maintain haemoglobin concentrations within -1.0 and +1.5 g/dl of the individual mean baseline haemoglobin and between 10 and 13 g/dl for 24 weeks. RESULTS: Mean change in haemoglobin from baseline to the evaluation period (weeks 21-24) was 0.13 g/dl (95% CI, 0.01, 0.25), which was not clinically relevant. An analysis by route of administration revealed that mean haemoglobin concentrations had increased by 0.58 g/dl (95% CI, 0.33, 0.82) in patients receiving i.v. darbepoetin alfa, and previously treated with i.v. rHuEpo, while remaining unchanged in s.c. patients (0.00 g/dl; 95% CI, -0.13, 0.13) previously treated by s.c. rHuEpo. In addition, there was a statistically significant decrease in mean weekly i.v. darbepoetin alfa dose requirements from 25.2 microg/week at baseline to 21.5 microg/week (P=0.004) during the evaluation period (-17.3%). Subcutaneous weekly dosage requirements increased slightly during the study period (20.8 to 22.7 microg/week; P=0.014). An i.v./s.c. dose ratio of 0.95 (95% CI, 0.78, 1.14) at evaluation confirms previous findings that dose requirements by the i.v. and s.c. routes were not different in patients treated with darbepoetin alfa. Haemoglobin concentrations were also effectively maintained in patients who received darbepoetin alfa once weekly and once every other week. Darbepoetin alfa was well tolerated. CONCLUSIONS: The treatment of renal anaemia in dialysis patients with unit doses of darbepoetin alfa effectively and safely maintains target haemoglobin concentrations with less frequent dosing. Dose requirements for darbepoetin alfa following i.v. and s.c. administration were not different. The results of this study demonstrate that darbepoetin alfa administered i.v. once weekly, or once every other week is an effective treatment regimen for haemodialysis patients with renal anaemia. PMID- 12543894 TI - Early prediction of response to intravenous iron supplementation by reticulocyte haemoglobin content and high-fluorescence reticulocyte count in haemodialysis patients. AB - BACKGROUND: Optimal response to recombinant human erythropoietin (rHuEpo) in haemodialysis (HD) patients requires provision of sufficient available iron. However, a balance between iron requirements and supplements remains a challenge in clinical practice. Reticulocyte parameters, i.e. reticulocyte haemoglobin content (CHr) and reticulocytes in a high-fluorescence intensity region (HFR), have been shown to be accurate predictors of iron-deficient erythropoiesis as compared with traditional markers. Therefore, the aim of this study was to appraise the diagnostic power of these two parameters in the early prediction of response to intravenous iron (IVFE) medications in HD patients receiving rHuEpo. METHODS: Sixty-five HD patients with a serum ferritin level of <500 microg/l and on rHuEpo therapy for >6 months were enrolled for IVFE supplementation (100 mg iron saccharate three times a week for 4 weeks, then 100 mg every 2 weeks for 5 months). Haemoglobin, haematocrit, serum ferritin, transferrin saturation, reticulocyte count, percentage of hypochromic red cells, CHr and HFR were measured before and following iron supplementation. Response was defined as a rise in haematocrit of >3% and/or a reduction in rHuEpo dose of >30% over the baseline values at the end of the study. RESULTS: Forty-two patients had a dramatic response to IVFE therapy with a 13.5% increase in mean haematocrit and a 38% reduction in rHuEpo dose at the end of the study (P<0.001). This paralleled a statistically significant rise in CHr and HFR (P<0.001). Univariate analyses showed that ferritin (P<0.010) and CHr (P<0.001) at baseline, changes in CHr (DeltaCHr(2W), P<0.001) and HFR (DeltaHFR(2W), P<0.010) at 2 weeks, as well as changes in CHr (DeltaCHr(4W), P<0.001) and HFR (DeltaHFR(4W), P<0.001) at 4 weeks, strongly correlated with response to IVFE supplementation. Stepwise discriminant analysis disclosed that DeltaCHr(4W) in conjunction with DeltaHFR(4W) exhibited an r(2) value of 0.531 (P<0.001) to predict response to IVFE therapy. Analyses by receiver operating characteristic curves and logistic regression further revealed that DeltaCHr(4W) at a cut-off value of >1.2 pg and DeltaHFR(4W) of >500/microl were more specific to the status of iron-deficient erythropoiesis following IVFE medications. Combined use of the two cut-off values allowed for the highest accuracy in the early prediction of the response to IVFE therapy, with a sensitivity of 96% and a specificity of 100%. CONCLUSIONS: Our study shows that changes in CHr and HFR at either 2 or 4 weeks are superior to the conventional erythrocyte and iron metabolism indices and may serve as reliable parameters to detect iron-deficient erythropoiesis in HD patients undergoing rHuEpo therapy. During aggressive IVFE treatment, early identification of non-responsiveness and subsequent discontinuation of treatment can avoid the inadvertent iron-related toxicity due to over-treatment. PMID- 12543895 TI - Prospective evaluation of failure modes in autogenous radiocephalic wrist access for haemodialysis. AB - INTRODUCTION: Radiocephalic wrist arteriovenous fistulae (RCAVF) are the primary and best option for vascular access for haemodialysis treatment. However, 10-24% of these AVFs fail due directly to thrombosis and non-maturation. In a prospective study, the failure modes of radiocephalic AVFs and the impact of surgical and interventional treatment on fistula outcome were investigated. METHODS: The rate of thrombosis and non-maturation was evaluated in 43 RCAVFs. The selection of RCAVF creation was made on preoperatively determined duplex parameters. Fistula function was evaluated post-operatively by clinical examination and non-invasively measured AVF blood flow. A policy of a liberal use of radiological and/or surgical revision of non-functioning RCAVFs was made on the basis of duplex measured blood flow and angiographically detected vessel stenosis. RESULTS: Primary fistula function was achieved in 26 of 43 patients (60%). Non-maturation and thrombosis occurred in 14 (33%) and three (7%) patients, respectively. A total of 12 interventions (PTA 6; surgery 6) were needed, resulting in salvage of eight RCAVFs (47%). The blood flow in functioning AVFs was significantly higher compared to non-functioning AVFs at 1 (754 vs 440 cc/min), 7 (799 vs 524 cc/min) and 42 days (946 vs 532 cc/min) post-operatively. At the end, 34 RCAVFs (79%) became functional as vascular access for haemodialysis treatment. CONCLUSION: Primary RCAVFs have a high rate of failure. An aggressive approach towards early interventional treatment of these non functional AVFs is worthwhile and leads to a considerable salvage rate. Early post-operative AVF flow measurement indicates the chance of successful maturation of RCAVF. PMID- 12543896 TI - Comparison of anthropometric equations for estimation of total body water in peritoneal dialysis patients. AB - BACKGROUND: Several formulae exist for estimating total body water (TBW). We aimed to assess their validity in peritoneal dialysis patients by comparison with TBW estimated by deuterium oxide dilution (TBW(D)). METHODS: We compared the equations of Chertow (TBW(Cher)), Chumlea (TBW(Chum)), Hume and Weyers (TBW(HW)), Johansson (TBW(J)), Lee (TBW(L)), Watson (TBW(W)) and TBW as 58% of body weight (TBW(0.58Wt)) with TBW(D) in 31 peritoneal dialysis (PD) patients and 32 controls. Estimates were compared with TBW(D) using Bland and Altman comparison. Extracellular water (ECW) was also estimated by sodium bromide dilution. RESULTS: In PD patients, mean TBW(D) was 35.04 (SD 7.84) l. Estimates were greater for TBW(Cher), TBW(Chum), TBW(HW), TBW(J) and TBW(0.58Wt). Mean TBW(L) and TBW(W) did not differ from TBW(D). Ninety-five percent limits of agreement (LOA) compared with TBW(D) (as a percentage of the mean) were similar for all of the different equations in PD patients (between +/-15.4 and +/-17.3%) except TBW(0.58Wt), which was far greater (+/-26.4%). In controls, mean TBW(D) was 37.03 (SD 6.63) l. Estimates were greater for TBW(Cher), TBW(Chum), TBW(HW), TBW(J) and TBW(0.58Wt). Mean TBW(L) and TBW(W) did not differ from TBW(D). Ninety-five percent LOA compared with TBW(D) (as a percentage of the mean) were similar for all equations in the controls, and closer than in PD patients (between +/-9.1 and +/-11.5%) except TBW(0.58Wt), which was again far greater than the other equations (+/ 28.1%). TBW(HW) - TBW(D) correlated with mean TBW (r=-0.412, P<0.05 in PD and r= 0.383, P<0.05 in controls). TBW(W) - TBW(D) (r=-0.539, P<0.005) correlated with mean TBW in PD. TBW(0.58Wt) - TBW(D) correlated with body mass index (BMI) (r=0.624, P<0.0001 in PD and r=0.829, P<0.0001 in controls) and ECW/TBW (r=0.406, P<0.05 in PD and r=0.411, P<0.02 in controls). CONCLUSIONS: Predictive equations were less accurate in PD than controls. TBW(0.58Wt) was most inaccurate, with systematic overestimation of TBW with increasing BMI and ECW/TBW. There were no differences in LOA with TBW(D) for the other equations within each group. PMID- 12543897 TI - Economic impact of extended time on peritoneal dialysis as a result of using polyglucose: the application of a Markov chain model to forecast changes in the development of the ESRD programme over time. AB - BACKGROUND: The use of polyglucose as a peritoneal dialysis (PD) fluid extends time on PD treatment. It is anticipated, therefore, that the share of patients treated with PD will be positively influenced. The relationship between extension of PD treatment time and an increase of the PD treatment share, however, is complex and needs further investigation. In this paper, a Markov chain model was applied to investigate the impact of extended time on PD treatment for the PD share in all dialysis patients in The Netherlands. Furthermore, the economic impact of the extended time on treatment (ETOT) was explored. METHODS: Scenarios were forecast over a 10 year period using aggregate data from the End-Stage Renal Registry in The Netherlands (Renine). Three scenarios were simulated in which the median PD technique survival was extended by 8, 10 and 12 months. Two other scenarios explored the impact of the combined effect of ETOT of 10 months together with a 10% and 20% increase of PD inflow shares. Reductions of costs to society due to ETOT were estimated using Dutch cost data on renal replacement therapies. RESULTS: PD share increases from 30.0% in the null scenario to 34.5% in the scenario with an ETOT of 10 months and an increased PD inflow share of 20%. The reduction in total costs to society of the renal replacement therapies is 0.96%. The average societal costs per discounted patient year for haemodialysis (HD) are 84 100 euros. For PD, these costs are 60 300 euros. A shift from HD to PD results in average cost savings of 28% per patient year. CONCLUSIONS: In view of high dialysis costs to society, a reduction of 0.96% can be considered to be relevant for healthcare policy makers. PMID- 12543898 TI - CMV infection of the renal allograft is much more common than the pathology indicates: a retrospective analysis of qualitative and quantitative buffy coat CMV-PCR, renal biopsy pathology and tissue CMV-PCR. AB - BACKGROUND: Quantitative blood polymerase chain reaction (PCR) for cytomegalovirus (CMV) is used to direct therapy in kidney transplant patients, but cytomegalic inclusions are rarely found in allograft renal biopsies even with an elevated serum creatinine and apparent CMV disease. The relationship between quantitative blood CMV and renal allograft pathology is unknown. METHODS: Thirteen biopsy samples were available for analysis from patients suspected of CMV disease, who had a buffy coat CMV-PCR drawn within 2-5 days of a renal allograft biopsy for an elevated creatinine. All were evaluated for CMV pathologically, by light microscopy, immunohistochemistry, in situ hybridization and tissue PCR. RESULTS: Qualitative and quantitative buffy coat CMV-PCR were positive in 10/13 (77%) patients. Tissue CMV-PCR was positive in five (50%) biopsies, including two with CMV inclusions and three with no inclusions. Quantitative buffy coat CMV-PCR levels did not correlate with detection of CMV inclusions in renal tissue. Paradoxically, quantitative buffy coat CMV-PCR was low (239 and 538 copies/microg of DNA) when CMV inclusions were detected. All five biopsies with acute rejection were associated with CMV viraemia and two of the five with allograft CMV inclusions. A quantitative buffy coat CMV-PCR of <100 copies/microg of DNA ruled out disease with CMV inclusions. CONCLUSIONS: CMV nephropathy is much more common than previously reported when sensitive techniques are used for detection in tissue. Acute rejection and CMV viraemia occur commonly together in patients at risk for CMV. Quantitative buffy coat CMV PCR does not correlate with the presence of CMV inclusions. These findings have implications for management of patients who have elevated serum creatinine and are at risk for CMV disease. PMID- 12543899 TI - Estimation of individual sensitivity to cyclosporin in children awaiting renal transplantation. AB - BACKGROUND: Optimal immunosuppressive drug therapy requires that efficacy be balanced against toxicity. We have performed in vitro assays of cyclosporin (CsA) efficacy in children awaiting renal transplantation. METHODS: Peripheral blood mononuclear cells (PBMC) from 13 children awaiting renal transplantation and 10 healthy paediatric controls ("responders") were incubated in the presence of CsA (0-250 ng/ml). Irradiated PBMC from a parent (prospective live donor) were cultured with those of the child in the presence of interleukin 2. Europium labelled, non-irradiated phytohaemagluttinin-stimulated target cells from the parent were added to the culture after 7 days incubation. Target cell lysis was quantified by time resolved fluorometry. CsA-mediated inhibition of target cell lysis was calculated and used to compare individual responses to the drug. Two colour flow cytometry was performed to identify activated subsets of lymphocytes at varying concentrations of CsA. RESULTS: Wide inter-individual variations in per cent lysis and per cent inhibition were observed in patients and controls. Immunophenotyping indicated expansion of CD8(+) and CD25(+) lymphocyte subsets following allo-stimulation that was inhibited by increasing concentrations of CsA. Eight out of 13 patients and four out of 10 controls were "sensitive" to CsA in vitro in that they achieved 50% inhibition of cell lysis (IC(50)) at low concentrations of the drug (<50 ng/ml). Eleven patients have subsequently received a renal transplant. Five out of seven of these patients with IC(50) <50 ng/ml have suffered problems with infection, nephrotoxicity and graft vasculopathy raising the possibility of "over-immunosuppression". CONCLUSION: The data imply a useful role for this model in the prediction of individual response to immunosuppression following allo-stimulation in the pre-transplant setting. PMID- 12543900 TI - Early experience with sildenafil for the treatment of erectile dysfunction in renal transplant recipients. AB - BACKGROUND: Erectile dysfunction (ED) is common in men with renal failure, but is not always alleviated following kidney transplant. The objective of the present study was to assess the feasibility in renal transplant patients of sildenafil citrate treatment, an agent with proven efficacy in the management of ED. METHODS: This was a phase IV, open, multicentre, 3 month, dose-escalation study. All patients meeting the inclusion criteria were prescribed a dose of 50 mg sildenafil at the first visit. Thereafter the dose could be increased to 100 mg or reduced to 25 mg based on efficacy or tolerability. The primary efficacy parameter assessed the ability of patients to achieve erections sufficient for intercourse and to maintain erections after penetration. Secondary endpoints assessed patient satisfaction with sildenafil and the effect of sildenafil on their quality of life. Patients were carefully monitored throughout the study for adverse events, interactions with immunosuppressive therapy and effect on graft function. RESULTS: The study included 50 patients in the intent-to-treat population. Sildenafil significantly improved patient's erection ability and the frequency of their erection maintenance. Analysis of the secondary efficacy parameters revealed that 66% of patients believed treatment had improved their erections. Patients reported improvements in their sexual life and partner relationships and a high level of satisfaction with treatment. There were no interactions between sildenafil and the immunosuppressive drugs and there was no significant adverse effect of sildenafil on graft function. CONCLUSIONS: Sildenafil is an effective and well-tolerated agent for the treatment of ED in renal transplant recipients. PMID- 12543901 TI - Kidney transplantation from related and unrelated living donors in a single German centre. AB - BACKGROUND: Organ transplantation began in 1954 with living related donation (LRD). Because of organ shortage from cadavers, unrelated kidney donation (LURD) has been proposed and shown to have good results despite complete HLA mismatching. This study aims to look at differences and similarities comparing LRD and LURD performed in our centre since the implementation of the German transplant law in 1997. METHODS: Between January 1997 and July 2001, 62 out of 112 potential living donors and their recipients were accepted. Immunosuppression consisted of triple therapy (steroids, cyclosporin, mycophenolate) in patients with three or fewer mismatches, or quadruple therapy including mono- or polyclonal antibody treatment in patients with four or more mismatches or cytotoxic antibodies. LRD and LURD groups were compared for number and type of rejections, complications and kidney function at the end of observation (median 15.5 months, range 1-50 months). RESULTS: Out of 112 pairs presenting, transplantation was performed in only 62 cases (55.4%). Reasons to deny transplantation were medical problems of the potential donors in 19, psychological problems in 13, recipient problems in seven and other reasons in 11 pairs. In 38 cases LRD transplantation and in 24 cases LURD transplantation was carried out. Recipient age was significantly lower in the LRD group (37.7+/-12.1 years) compared with the LURD group (53.6+/-7.8 years). Mean donor age was 49.7+/ 9.2 years in the LRD group and 50.3+/-9.1 years in the LURD group (ns). The number of mismatches was lower in LRD (2.1+/-1) than in LURD (4.4+/-0.9) (P=0.001) transplantation. The acute rejection rate was similar in both groups (52.2 vs 54.2%). OKT3 and tacrolimus rescue therapy for more severe rejections was more often applied in the LRD group but the difference did not reach the level of significance. There were more infectious complications in LURD transplantation (66.7 vs 36.4%, P=0.036) and a trend towards more surgical complications in LRD transplantation (28.9 vs 8.3%, P=0.062). One graft was lost due to transplant artery thrombosis and one recipient died 4 months after transplantation subsequent to cerebral ischaemia. Both patients belonged to the LRD group. Creatinine values at the end of observation time were 1.76+/-0.6 mg/dl in the LRD group and 1.62+/-0.5 mg/dl in the LURD group (ns). CONCLUSION: Although kidney transplantation from unrelated donors was performed with a lower HLA match and although the recipients were older, the results are equivalent to living related transplantation. Therefore, kidney transplantation from emotionally related living donors represents a valuable option for patients with end-stage renal disease. Careful selection of donors and recipients is a prerequisite of success. PMID- 12543902 TI - Invasive urothelial carcinoma after exposure to Chinese herbal medicine containing aristolochic acid may occur without severe renal failure. PMID- 12543903 TI - Leukocytoclastic vasculitis and interstitial nephritis with carbimazole treatment. PMID- 12543904 TI - Adherence of tunnelled haemodialysis catheter to superior vena caval stent: successful percutaneous removal. PMID- 12543905 TI - The man who gained a stone. PMID- 12543906 TI - Triangular face and vascular malformation. The links to renal failure. PMID- 12543907 TI - The importance of a histology-based diagnosis of interstitial nephropathy in two patients with renal insufficiency. PMID- 12543908 TI - Kidney at risk: 11-year course of renal artery stenosis. PMID- 12543909 TI - Familial phosphoglycerate kinase deficiency associated with rhabdomyolysis and acute renal failure: abnormality in mRNA splicing? PMID- 12543910 TI - Malnutrition, chronic inflammation and atherosclerosis in dialysis patients. PMID- 12543911 TI - On-line haemodiafiltration decreases serum TNFalpha levels in haemodialysis patients. PMID- 12543912 TI - Sirolimus and lymphocele formation after kidney transplantation: an immunosuppressive medication as co-factor for a surgical problem? PMID- 12543914 TI - Cytolethal distending toxin B gene (cdtB) homologues in taxa 2, 3 and 8 and in six canine isolates of Helicobacter sp. flexispira. AB - The presence of the cytolethal distending toxin B gene (cdtB) was examined in eight Helicobacter sp. flexispira reference strains, Helicobacter trogontum ATCC 700114(T) and 12 Finnish porcine H. trogontum strains and canine flexispira isolates. Part of the cdtB gene was amplified by PCR with degenerate primers VAT2 and DHF1, cloned and sequenced. The presence/absence of the cdtB gene as determined by PCR was confirmed by Southern hybridization and toxin production by HeLa cell-line experiments. PCR amplification resulted in approximately 700 bp fragments from Helicobacter sp. flexispira taxa 2 (ATCC 49314), 3 (ATCC 49320) and 8 (ATCC 43880, ATCC 49308, ATCC 43879), from six canine isolates as well as from the control strains Helicobacter bilis and Helicobacter hepaticus. The hybridization patterns of HaeIII-, HindIII- and AseI-digested chromosomal DNA confirmed the results of the PCR experiments. The cdtB-positive strains had effects ranging from weak to strong on HeLa cell cultures. PCR amplification from the reference strains Helicobacter sp. flexispira taxa 1 (ATCC 43968), 4 (ATCC 49310) and 5 (ATCC 43966) and H. trogontum (ATCC 700114(T)), and also six of the Finnish strains, was unsuccessful. No toxic effect on HeLa cells was evident when bacterial suspensions of PCR-negative strains were used for toxicity assay. Our results are in accordance with previous observations that the cdtB gene is not present in all Helicobacter species. Further, the presence/absence of the cdtB gene in Helicobacter sp. flexispira strains was in accordance with recent taxonomic analysis of the same strains, which suggests that it could serve as a useful marker in Helicobacter taxonomy. PMID- 12543915 TI - Role of staphylococcal enterotoxin A in a fatal case of endocarditis. AB - A young female with no identifiable risk factors developed rapid, overwhelming Staphylococcus aureus endocarditis. Despite rapid sterilization of the blood and the mitral valve with optimal antimicrobials, she had persistent septic shock. In order to investigate this, the toxin-producing capacity of the infecting strain and the patient's ability to produce antibodies were determined. The strain produced high levels of both alpha-toxin and staphylococcal enterotoxin A (SEA), whilst the patient responded with modestly high levels of antibodies to alpha toxin and low-normal levels to SEA. The patient was most probably susceptible to the actions of SEA and developed a toxic-shock-syndrome-like disease that further aggravated her valvular dysfunction. This case illustrates that optimal antimicrobial therapy alone is not sufficient treatment in patients with persistent toxic shock and that there is a need to evaluate immunomodulatory strategies in such patients. PMID- 12543916 TI - Manganese-dependent regulation of the endocarditis-associated virulence factor EfaA of Enterococcus faecalis. AB - There is increasing recognition of the emerging role of manganese regulation and acquisition in some pathogenic bacteria. Expression of the Enterococcus faecalis endocarditis-associated virulence factor EfaA is induced by growth in serum. It is demonstrated here that expression of the efaCBA operon encoding a putative ABC type transporter is regulated by Mn(2+). Transcription of efaCBA and EfaA production were repressed in Mn(2+)-supplemented medium. A Mn(2+)-responsive transcriptional regulator, EfaR, sharing 27 % identity with the Corynebacterium diphtheriae diphtheria toxin repressor (DtxR), was identified. In the presence of Mn(2+), EfaR protein bound in vitro to the efaC promoter region. Analysis of the E. faecalis V583 genome revealed ten additional putative EfaR-binding sites, suggesting that manganese availability could have a broader regulatory role in infection. The results identify a new Mn(2+)-sensing regulator in enterococci that regulates the expression of a virulence factor implicated in enterococcal endocarditis. PMID- 12543917 TI - Immune response to native NadA from Neisseria meningitidis and its expression in clinical isolates in Brazil. AB - A mAb against the NadA protein from Neisseria meningitidis strain 3006 (serosubtype B : 2b : P1.2 : P5.2,8) demonstrated strong bactericidal activity against Brazilian epidemic serogroup B strain N44/89 (B : 4,7 : P1.19,15 : P5.5,7) and a serogroup C strain, IMC 2135 (C : 2a : P1.5,2), but not against another serogroup C strain, N1002/90 (C : 2b : P1.3 : P5.8). The immunogenicity of native NadA in an outer-membrane vesicle (OMV) preparation was also tested. Serum from mice immunized with OMV from serogroup B strain N44/89, which contains the NadA protein, showed bactericidal activity against serogroup B and C strains possessing NadA. In dot-blot analysis of 100 serogroup B and 100 serogroup C isolates from Brazilian patients, the mAb to NadA recognized about 60 % of the samples from both serogroups. The molecular mass of the NadA protein from strain N44/89 determined by mass spectrometry was 37 971 Da and the peptide sequences were identical to those of NadA from N. meningitidis strain MC58. PMID- 12543918 TI - Low-stringency single specific primer PCR for identification of Leptospira. AB - Thirty-five Leptospira serovars from the species Leptospira interrogans, Leptospira borgpetersenii, Leptospira santarosai, Leptospira kirschneri, Leptospira weilii, Leptospira biflexa and Leptospira meyeri were characterized by the low-stringency single specific primer PCR (LSSP-PCR) technique. LSSP-PCR analysis was performed to detect DNA polymorphisms in a 285 bp DNA fragment amplified from genomic DNA with G1 and G2 selected primers. Similar LSSP-PCR profiles were obtained for serovars from the same genomic species, while serovars from non-related species produced distinct multiband patterns. Based on the data from sequence analysis, all genomic fragments amplified with G1 and G2 primers from distinct serovars of Leptospira were 285 bp in length, with nucleotide variation observed most frequently among different genomic species. The simplicity and accuracy of the LSSP-PCR technique were found to be suitable for identification of Leptospira species. PMID- 12543919 TI - Granulicatella adiacens and Abiotrophia defectiva bacteraemia characterized by 16S rRNA gene sequencing. AB - Traditionally, the identification, epidemiology and spectrum of clinical diseases caused by Granulicatella adiacens and Abiotrophia defectiva are dependent upon their phenotypic characterization. During a 6-year period (July 1995-June 2001), seven and two alpha-haemolytic streptococci were identified as G. adiacens and A. defectiva, respectively, by 16S rRNA gene sequencing. Three patients with haematological malignancies and neutropenic fever had primary bacteraemia. Three patients with valvular problems or congenital heart disease had infective endocarditis. A patient with ischemic heart disease and cerebrovascular accident had infected aortic atheroma with dissection. A patient with recurrent pyogenic cholangitis had acute cholangitis and a patient with polypoid cystitis and benign prostatic hypertrophy had acute prostatitis. Four of the nine patients died, including all three with G. adiacens infective endocarditis or infected atheroma. For the seven G. adiacens isolates, the API 20 STREP system successfully identified one and five isolates as G. adiacens with >95 % and 80-90 % confidence, respectively, whereas the Vitek System (GPI) and ATB Expression system (ID32 STREP) successfully identified none and one isolate as G. adiacens. Of the two A. defectiva isolates, none of the three systems successfully identified either of them as A. defectiva. 16S rRNA gene sequencing is the technique of choice for identifying G. adiacens and A. defectiva, and early surgical intervention should be considered when G. adiacens endocarditis is diagnosed. PMID- 12543920 TI - Characterization of class I integrons in clinical strains of Salmonella enterica subsp. enterica serovars Typhimurium and Enteritidis from Norwegian hospitals. AB - The characterization of integrons and their promoters in 156 antibiotic-resistant clinical isolates of the important zoonotic pathogens Salmonella enterica subsp. enterica serovars Typhimurium (S. Typhimurium) and Enteritidis (S. Enteritidis) from Norwegian hospitals were performed. Integrons were found in 64 of 66 S. Typhimurium isolates (97 %) and in 20 of 90 S. Enteritidis isolates (22.2 %) with the following sizes; 650, 1000, 1200, 1500, 1600, 1700, 2000 and 2100 bp. The integrons were further sequenced and the aadA1, aadA2, aadA5, aadB, pse-1, catB3, oxa1, dfrA1, dfrA12 and dfrA17 genes, as well as a fragment of the sat1 gene, were found embedded in cassettes. An internal fragment of the purG gene was additionally found as an artefact PCR amplicon. PMID- 12543921 TI - Independent subsets of amplified fragments from the genome of Neisseria meningitidis identify the same invasive clones of ET37 and ET5. AB - To determine whether fluorescent amplified-fragment length polymorphism (FAFLP) analysis is an unbiased genome sampling technique, data were analysed from three different primer combinations, amplifying three independent fragment subsets from 123 isolates of Neisseria meningitidis. Using these data, dendrograms were generated with near-identical topologies that identified the same invasive clones of ET37 and ET5 and also identified the same outbreak clusters. PMID- 12543922 TI - Antibiotic resistance among verocytotoxigenic Escherichia coli (VTEC) and non VTEC isolated from domestic animals and humans. AB - Two hundred verocytotoxigenic and 216 non-verocytotoxigenic Escherichia coli (VTEC and non-VTEC), isolated from a variety of sources were tested for their resistances to 11 antimicrobial agents. The strains included isolates from domestic food animals and both symptomatic and asymptomatic infections in man. A much higher level of resistance was found among the non-VTEC than among the VTEC, regardless of source. The resistant VTEC isolated from animals were predominantly from specimens associated with sick animals. Antibiotic resistance was detected in only four of the 59 (6.8 %) VTEC of human origin, whereas more of the human non-VTEC possessed antibiotic resistance determinants. It was particularly noteworthy that 24/87 (28 %) strains isolated from healthy babies, who had neither contact with antibiotics nor had gastrointestinal symptoms for at least 2 weeks prior to the specimen being taken, were resistant to one or more of the antibiotics tested. PMID- 12543923 TI - Species prevalence and antibacterial resistance of enterococci isolated in Kuwait hospitals. AB - This study investigated the species prevalence and antibacterial resistance among enterococci isolated in Kuwait hospitals. They consisted of 415 isolates of Enterococcus faecalis (85.3 %), Enterococcus faecium (7.7 %), Enterococcus casseliflavus (4.0 %), Enterococcus avium (1.2 %), Enterococcus durans (1.0 %), Enterococcus gallinarium (0.5 %) and Enterococcus bovis (0.2 %) isolated from urine (36.6 %), blood (10.4 %), wound swabs (11.0 %), stool samples (12.0 %), high vaginal swabs (9.0 %), endocervical swabs (3.0 %) and miscellaneous sources (18.0 %). All of them were susceptible to linezolid. Fifty-two (12.5 %) isolates were ampicillin resistant but none of them produced beta-lactamase. They were resistant to erythromycin (63.3 %), tetracycline (60.5 %), ciprofloxacin (40.0 %), chloramphenicol (28.0 %), vancomycin (2.6 %), and teicoplanin (2.6 %). Fourteen, 19 and 20 % of them expressed high-level resistance to gentamicin, kanamycin and streptomycin, respectively. All of the vancomycin-resistant strains carried the vanA phenotype and genotype. There was no evidence of clonal spread of the vancomycin-resistant isolates. PMID- 12543924 TI - Antifungal properties of 5-hydroxytryptamine (serotonin) against Candida species in vitro. AB - In this study the minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) of 5-hydroxytryptamine (5 HT, serotonin) against clinical isolates of Candida albicans (n = 11), Candida glabrata (n = 9), Candida tropicalis (n = 10) and Candida parapsilosis (ATCC 22019) using a broth microdilution test were investigated. In addition, it was examined whether delayed regrowth as a post-antifungal effect results following short exposure to 5 HT. 5 HT showed antifungal activity towards all isolates of Candida spp. The isolates yielded comparable MIC and MFC values of 5 HT in the range 0.91-7.34 mM and 1.83-14.68 mM, respectively. A lag in regrowth was dependent on the concentration tested. Treatment for 3 h at concentrations of 5 HT below and equipotent to the MFC resulted in a delayed regrowth of 8-12 h for isolates of Candida spp. In conclusion, these in vitro studies clearly demonstrate antifungal effects of 5 HT. Identifying the mode of action could be of great help in developing and researching new antifungal drugs. PMID- 12543925 TI - Neisseria meningitidis phenotypic markers and septicaemia, disease progress and case-fatality rate of meningococcal disease: a 20-year population-based historical follow-up study in a Danish county. AB - The incidence rate (IR) and case-fatality rate (CFR) of meningococcal disease increased during the late 1980s and early 1990s in North Jutland County, Denmark. We examined the hypothesis that phenotypic markers of Neisseria meningitidis are predictors of septicaemia with or without meningitis, rapid disease progress and fatal outcome of meningococcal disease and we studied whether changes in IR and CFR over time might be related to emergence or spread of certain phenotypes. This follow-up study was based on a complete registration of 413 cases of meningococcal disease in North Jutland County during 1980-99. Phenotypic markers included serogroup, serotype and serosubtype. A complete phenotype was available for 315 cases (76 %); 100 (32 %) strains were phenotype B : 15 : P1.7,16 and 31 (10 %) were C : 2a : P1.2,5. Septicaemia without meningitis was less common in cases with B : 15 : P1.7,16 and C : 2a : P1.2,5 strains. No association was found between phenotype and rapid disease progress. The overall CFR was 12 %. An increased CFR was associated with phenotypes B : 15 : P1.7,16 [odds ratio (OR) 2.8, 95 % confidence interval (CI) 1.2-18.5] and C : 2a : P1.2,5 (OR 5.2, 95 % CI 1.6-16.4) when compared with other phenotypes. The prevalence of B : 15 : P1.7,16 strains increased gradually during the study period and the CFR increased from 8 % during 1980-89 to 19 % during 1990-99, although the CFR for other phenotypes also increased. The CFR for C : 2a : P1.2,5 remained high ( approximately 20 %), but the contribution of this phenotype to the overall CFR decreased during the study period. In conclusion, phenotypes B : 15 : P1.7,16 and C : 2a : P1.2,5 were predictors of an increased CFR. The high prevalence of phenotype B : 15 : P1.7,16 contributed to increased overall IR and CFR during 1990-99. PMID- 12543926 TI - Molecular epidemiology and characteristics of Corynebacterium diphtheriae and Corynebacterium ulcerans strains isolated in Italy during the 1990s. AB - Five cases of diphtheria were reported in Italy between January 1990 and June 2001. Three cases were confirmed microbiologically by the isolation of toxigenic Corynebacterium diphtheriae (two cases) and Corynebacterium ulcerans (one case). Over the same period, 11 cases of non-toxigenic C. diphtheriae infection were reported to the Italian Public Health Institute, from which the causative organism was isolated from a skin infection in one case and from the throat in the other ten. Seven of the throat isolates were associated with fever, severe pharyngitis and tonsillitis and were all biotype gravis. Because there are no standardized breakpoints, the antimicrobial sensitivities of C. diphtheriae were determined in accordance with the National Committee for Clinical Laboratory Standards guidelines for Streptococcus spp. other than Streptococcus pneumoniae. MICs for penicillin ranged between 0.125 and 0.250 mg l(-1) and 7 out of 11 strains had a minimal bactericidal concentration (MBC)/MIC ratio >or= 32. All strains were sensitive to clindamycin (MIC /=50% of a single residue type are surveyed in this work. Both eukaryote and prokaryote proteomes are investigated with emphasis on the eukaryotes. Very large numbers of such sequences are found in all organisms surveyed. It is found that eukaryotes possess far more simple sequences per protein than do the prokaryotes. Prokaryotes display a linear relationship between number of proteins containing simple sequences and proteome size, whereas it is not clear that such a relationship holds for eukaryotes. Strikingly, it is found that each eukaryote possesses its own unique distribution of simple sequences. Within those distributions it is found that simple sequences enriched in certain residue types are clearly favored, whereas others are just as clearly discriminated against. The preferences observed are not correlated with residue occurrence. An analysis of classes of proteins of known function suggests that simple sequence occurrence and distribution may be related to protein function. Based upon this analysis, the large number of simple sequences found above that would be expected from a simple statistical model, plus the known functional importance of numerous such sequences, it is postulated that eukaryotes have evolved to not only tolerate large numbers of simple sequences but also to require them. PMID- 12543935 TI - Poxviruses and immune evasion. AB - Large DNA viruses defend against hostile assault executed by the host immune system by producing an array of gene products that systematically sabotage key components of the inflammatory response. Poxviruses target many of the primary mediators of innate immunity including interferons, tumor necrosis factors, interleukins, complement, and chemokines. Poxviruses also manipulate a variety of intracellular signal transduction pathways such as the apoptotic response. Many of the poxvirus genes that disrupt these pathways have been hijacked directly from the host immune system, while others have demonstrated no clear resemblance to any known host genes. Nonetheless, the immunological targets and the diversity of strategies used by poxviruses to disrupt these host pathways have provided important insights into diverse aspects of immunology, virology, and inflammation. Furthermore, because of their anti-inflammatory nature, many of these poxvirus proteins hold promise as potential therapeutic agents for acute or chronic inflammatory conditions. PMID- 12543937 TI - IP policies and serving the public. PMID- 12543936 TI - The regulatory role of Valpha14 NKT cells in innate and acquired immune response. AB - A novel lymphocyte lineage, Valpha14 natural killer T (NKT) cells, is now well established as distinct from conventional alphabeta T cells. Valpha14 NKT cells express a single invariant Valpha14 antigen receptor that is essential for their development. Successful identification of a specific ligand, alpha galactosylceramide(alpha-GalCer), and the establishment of gene-manipulated mice with selective loss of Valpha14 NKT cells helped elucidate the remarkable functional diversity of Valpha14 NKT cells in various immune responses such as host defense by mediating anti-nonself innate immune reaction, homeostatic regulation of anti-self responses, and antitumor immunity. PMID- 12543938 TI - Planetary exploration. Europe's comet chaser put on hold following launcher failure. PMID- 12543939 TI - Bioterrorism. Go slow with smallpox shots, panel says. PMID- 12543940 TI - Bioinformatics. Private pact ends the DNA data war. PMID- 12543941 TI - Bioweapons. Plague of lies lands Texas scientist in jail. PMID- 12543942 TI - Scientific advice. Spanish researchers vent anger over handling of oil spill. PMID- 12543943 TI - Paleontology. Four-winged dinos create a flutter. PMID- 12543944 TI - Astronomy. Fire destroys historic Australian observatory. PMID- 12543945 TI - Astrophysics. Stanford gets serious about space physics. PMID- 12543946 TI - Russia. Putin aims to turn science cities into silicon steppes. PMID- 12543947 TI - Embryonic stem cells. Key questions loom over effort to energize research. PMID- 12543948 TI - Gene therapy. Seeking the cause of induced leukemias in X-SCID trial. PMID- 12543949 TI - Neuroscience. Tapping the mind. PMID- 12543950 TI - Neuroscience. Power to the paralyzed. PMID- 12543951 TI - American Astronomical Society meeting. Through a lens, deeply. PMID- 12543952 TI - American Astronomical Society meeting. A tsunami of hot Jupiters? PMID- 12543953 TI - American Astronomical Society meeting. Stars behaving badly. PMID- 12543954 TI - Structural biology. Propelled by recent advances, NMR moves into the fast lane. PMID- 12543955 TI - Society for Integrative and Comparative Biology meeting. Shedding light on avian iridescence. PMID- 12543956 TI - Society for Integrative and Comparative Biology meeting. Seeing the invisible. PMID- 12543957 TI - Society for Integrative and Comparative Biology meeting. Lean times, lean gut. PMID- 12543958 TI - Communication. Science and the semantic web. PMID- 12543959 TI - Development. The bills of qucks and duails. PMID- 12543960 TI - Evolution. Haploid superiority. PMID- 12543961 TI - Chemistry. Shaped laser pulses as reagents. PMID- 12543963 TI - Geophysics. An ensemble view of Earth's inner core. PMID- 12543962 TI - Transcription. Puffing with PARP. PMID- 12543964 TI - Biomedicine. Nota bene: weathering the big chill. PMID- 12543965 TI - Information storage using supramolecular surface patterns. PMID- 12543966 TI - Deciphering the reaction dynamics underlying optimal control laser fields. AB - Femtosecond high-resolution pump-probe experiments have been used together with theoretical ab initio quantum calculations and wave packet dynamics simulations to decode an optimal femtosecond pulse that is generated from adaptive learning algorithms. This pulse is designed to maximize the yield of the organometallic ion CpMn(CO)3 while hindering the competing fragmentation. The sequential excitation and ionization of the target ion are accomplished by an optimized field consisting of two dominant subpulses with optimal frequencies and time delays. PMID- 12543967 TI - High-accuracy ab initio rotation-vibration transitions for water. AB - The spectrum of water vapor is of fundamental importance for a variety of processes, including the absorption and retention of sunlight in Earth's atmosphere. Therefore, there has long been an urgent need for a robust and accurate predictive model for this spectrum. In our work on the high-resolution spectrum of water, we report first-principles calculations that approach experimental accuracy. To achieve this, we performed exceptionally large electronic structure calculations and considered a variety of effects, including quantum electrodynamics, which have routinely been neglected in studies of small many-electron molecules. The high accuracy of the resulting ab initio procedure is demonstrated for the main isotopomers of water. PMID- 12543968 TI - Vibrationally resolved fluorescence excited with submolecular precision. AB - Tunneling electrons from a scanning tunneling microscope (STM) were used to excite photon emission from individual porphyrin molecules adsorbed on an ultrathin alumina film grown on a NiAl(110) surface. Vibrational features were observed in the light-emission spectra that depended sensitively on the different molecular conformations and corresponding electronic states obtained by scanning tunneling spectroscopy. The high spatial resolution of the STM enabled the demonstration of variations in light-emission spectra from different parts of the molecule. These experiments realize the feasibility of fluorescence spectroscopy with the STM and enable the integration of optical spectroscopy with a nanoprobe for the investigation of single molecules. PMID- 12543969 TI - Long-range correlations in the diffuse seismic coda. AB - The late seismic coda may contain coherent information about the elastic response of Earth. We computed the correlations of the seismic codas of 101 distant earthquakes recorded at stations that were tens of kilometers apart. By stacking cross-correlation functions of codas, we found a low-frequency coherent part in the diffuse field. The extracted pulses have the polarization characteristics and group velocities expected for Rayleigh and Love waves. The set of cross correlations has the symmetries of the surface-wave part of the Green tensor. This seismological example shows that diffuse waves produced by distant sources are sufficient to retrieve direct waves between two perfectly located points of observation. Because it relies on general properties of diffuse waves, this result has potential applications in other fields. PMID- 12543970 TI - A nebular origin for chondritic fine-grained phyllosilicates. AB - Hydrated minerals occur in accretionary rims around chondrules in CM chondrites. Previous models suggested that these phyllosilicates did not form by gas-solid reactions in the canonical solar nebula. We propose that chondrule-forming shock waves in icy regions of the nebula produced conditions that allowed rapid mineral hydration. The time scales for phyllosilicate formation are similar to the time it takes for a shocked system to cool from the temperature of phyllosilicate stability to that of water ice condensation. This scenario allows for simultaneous formation of chondrules and their fine-grained accretionary rims. PMID- 12543971 TI - Robust normal mode constraints on inner-core anisotropy from model space search. AB - A technique for searching full model space that was applied to measurements of anomalously split normal modes showed a robust pattern of P-wave and S-wave anisotropy in the inner core. The parameter describing P-wave anisotropy changes sign around a radius of 400 kilometers, whereas S-wave anisotropy is small in the upper two-thirds of the inner core and becomes negative at greater depths. Our results agree with observed travel-time anomalies of rays traveling at epicentral distances varying from 150 degrees to 180 degrees. The models may be explained by progressively tilted hexagonal close-packed iron in the upper half of the inner core and could suggest a different iron phase in the center. PMID- 12543972 TI - An evolutionary advantage of haploidy in large yeast populations. AB - Although seed plants and multicellular animals are predominantly diploid, the prominence of diploidy varies greatly among eukaryote life cycles, and no general evolutionary advantage of diploidy has been demonstrated. By doubling the copy number of each gene, diploidy may increase the rate at which adaptive mutations are produced. However, models suggest that this does not necessarily accelerate adaptation by diploid populations. We tested model predictions regarding rates of adaptation using asexual yeast populations. Adaptive mutations were on average partially recessive. As predicted, diploidy slowed adaptation by large populations but not by small populations. PMID- 12543973 TI - Tracheal respiration in insects visualized with synchrotron x-ray imaging. AB - Insects are known to exchange respiratory gases in their system of tracheal tubes by using either diffusion or changes in internal pressure that are produced through body motion or hemolymph circulation. However, the inability to see inside living insects has limited our understanding of their respiration mechanisms. We used a synchrotron beam to obtain x-ray videos of living, breathing insects. Beetles, crickets, and ants exhibited rapid cycles of tracheal compression and expansion in the head and thorax. Body movements and hemolymph circulation cannot account for these cycles; therefore, our observations demonstrate a previously unknown mechanism of respiration in insects analogous to the inflation and deflation of vertebrate lungs. PMID- 12543974 TI - Chromatin loosening by poly(ADP)-ribose polymerase (PARP) at Drosophila puff loci. AB - Steroid response and stress-activated genes such as hsp70 undergo puffing in Drosophila larval salivary glands, a local loosening of polytene chromatin structure associated with gene induction. We find that puffs acquire elevated levels of adenosine diphosphate (ADP)-ribose modified proteins and that poly(ADP) ribose polymerase (PARP) is required to produce normal-sized puffs and normal amounts of Hsp70 after heat exposure. We propose that chromosomal PARP molecules become activated by developmental or environmental cues and strip nearby chromatin proteins off DNA to generate a puff. Such local loosening may facilitate transcription and may transiently make protein complexes more accessible to modification, promoting chromatin remodeling during development. PMID- 12543975 TI - Phosphorus limitation of coastal ecosystem processes. AB - Primary production in coastal wetlands is conventionally thought to be limited by nitrogen. Although the plant community in a pristine salt marsh was found to be limited primarily by nitrogen availability, the bacterial community in the soil was limited by phosphorus. Hence, in coastal wetlands, and possibly in many ecosystems, individual trophic groups may respond differently to nitrogen and phosphorus loading. Phosphorus limitation of the growth of nitrogen-transforming bacteria will affect carbon fixation, storage, and release mediated by plants, a result that has important implications for ecosystem management. PMID- 12543976 TI - The cellular and molecular origins of beak morphology. AB - Cellular and molecular mechanisms underlying differences in beak morphology likely involve interactions among multiple embryonic populations. We exchanged neural crest cells destined to participate in beak morphogenesis between two anatomically distinct species. Quail neural crest cells produced quail beaks in duck hosts and duck neural crest produced duck bills in quail hosts. These transformations involved morphological changes to non-neural crest host beak tissues. To achieve these changes, donor neural crest cells executed autonomous molecular programs and regulated gene expression in adjacent host tissues. Thus, neural crest cells are a source of molecular information that generates interspecific variation in beak morphology. PMID- 12543977 TI - Functional mapping of the primate auditory system. AB - Cerebral auditory areas were delineated in the awake, passively listening, rhesus monkey by comparing the rates of glucose utilization in an intact hemisphere and in an acoustically isolated contralateral hemisphere of the same animal. The auditory system defined in this way occupied large portions of cerebral tissue, an extent probably second only to that of the visual system. Cortically, the activated areas included the entire superior temporal gyrus and large portions of the parietal, prefrontal, and limbic lobes. Several auditory areas overlapped with previously identified visual areas, suggesting that the auditory system, like the visual system, contains separate pathways for processing stimulus quality, location, and motion. PMID- 12543978 TI - Extended longevity in mice lacking the insulin receptor in adipose tissue. AB - Caloric restriction has been shown to increase longevity in organisms ranging from yeast to mammals. In some organisms, this has been associated with a decreased fat mass and alterations in insulin/insulin-like growth factor 1 (IGF 1) pathways. To further explore these associations with enhanced longevity, we studied mice with a fat-specific insulin receptor knockout (FIRKO). These animals have reduced fat mass and are protected against age-related obesity and its subsequent metabolic abnormalities, although their food intake is normal. Both male and female FIRKO mice were found to have an increase in mean life-span of approximately 134 days (18%), with parallel increases in median and maximum life spans. Thus, a reduction of fat mass without caloric restriction can be associated with increased longevity in mice, possibly through effects on insulin signaling. PMID- 12543979 TI - Disruption of transforming growth factor-beta signaling in ELF beta-spectrin deficient mice. AB - Disruption of the adaptor protein ELF, a beta-spectrin, leads to disruption of transforming growth factor-beta (TGF-beta) signaling by Smad proteins in mice. Elf-/- mice exhibit a phenotype similar to smad2+/-/smad3+/- mutant mice of midgestational death due to gastrointestinal, liver, neural, and heart defects. We show that TGF-beta triggers phosphorylation and association of ELF with Smad3 and Smad4, followed by nuclear translocation. ELF deficiency results in mislocalization of Smad3 and Smad4 and loss of the TGF-beta-dependent transcriptional response, which could be rescued by overexpression of the COOH terminal region of ELF. This study reveals an unexpected molecular link between a major dynamic scaffolding protein and a key signaling pathway. PMID- 12543980 TI - Dynamics of the hippocampus during encoding and retrieval of face-name pairs. AB - The medial temporal lobe (MTL) is critical in forming new memories, but how subregions within the MTL carry out encoding and retrieval processes in humans is unknown. Using new high-resolution functional magnetic resonance imaging (fMRI) acquisition and analysis methods, we identified mnemonic properties of different subregions within the hippocampal circuitry as human subjects learned to associate names with faces. The cornu ammonis (CA) fields 2 and 3 and the dentate gyrus were active relative to baseline only during encoding, and this activity decreased as associations were learned. Activity in the subiculum showed the same temporal decline, but primarily during retrieval. Our results demonstrate that subdivisions within the hippocampus make distinct contributions to new memory formation. PMID- 12543981 TI - Spectroscopic determination of the OH- solvation shell in the OH-.(H2O)n clusters. AB - There has been long-standing uncertainty about the number of water molecules in the primary coordination environment of the OH- and F- ions in aqueous chemistry. We report the vibrational spectra of the OH-.(H2O)n and F-.(H2O)n clusters and interpret the pattern of OH stretching fundamentals with ab initio calculations. The spectra of the cold complexes are obtained by first attaching weakly bound argon atoms to the clusters and then monitoring the photoinduced evaporation of these atoms when an infrared laser is tuned to a vibrational resonance. The small clusters (n 90 dB, SD <20%) after acoustic neuroma excision (n = 7), meningitis (n = 1), sudden sensorineural hearing loss (n = 1), and sudden sensorineural hearing loss with chronic suppurative otitis media (n = 1). Entry criteria included normal hearing in the contralateral ear (pure tone average <25 dB, SD >80%). INTERVENTION: Subjects were fitted with contralateral routing of offside signal amplification devices for 1 month and tested with contralateral routing of offside signal before mastoid implantation of the deaf ear, fitting, and testing for bone anchored hearing aid. OUTCOME MEASURES: Subjects' assessment of experience with their devices and patterns of use, 2) source azimuth identification in noise test, and 3) speech discrimination in quiet and in noise under conditions of noise-front, noise-to-normal-ear, and noise-to-deaf-ear. RESULTS: There was consistent satisfaction with bone anchored hearing aid implantation and amplification, and poor acceptance of contralateral routing of offside signal amplification. Sound localization was poor at baseline and with both bone anchored hearing aid and contralateral routing of offside signal. Relative to baseline, contralateral routing of offside signal and bone anchored hearing aid produced significantly better speech recognition in noise under most conditions. The bone anchored hearing aid enabled significantly better speech recognition than contralateral routing of offside signal in quiet and in a composite of noise conditions. The advantages may relate to averting the interference of speech signals delivered to the better ear, as occurs with conventional contralateral routing of offside signal amplification. CONCLUSIONS: Preliminary data in subjects with normal monaural hearing indicate that vibromechanical stimulation with the bone anchored hearing aid overcomes some of the negative head shadow effects in unilateral deafness. The bone anchored hearing aid system, when placed on the side of a deaf ear, yields greater benefit in subjects with normal monaural hearing than does contralateral routing of offside signal amplification. It seems that this rehabilitative approach can expand the sound field of monaural listeners in further enhancing speech understanding. Observations suggest that further understanding of bone conduction as implemented in transcranial stimulation will guide further options for patients with monaural hearing. Longer follow-up will help to determine whether communicative skill improvements with the bone anchored hearing aid outweigh the disadvantages of implantation surgery, costs, and device maintenance. PMID- 12544033 TI - Auditory brainstem implant in bilateral and completely ossified cochleae. AB - OBJECTIVE: To report and evaluate the results of auditory brainstem implantation in a case of postmeningitis hearing loss with totally ossified cochleae on both sides. STUDY DESIGN: Case report. SETTING: Tertiary referral center. PATIENT: A 49-year-old man was referred to the authors' department for bilateral total hearing loss subsequent to bacterial meningitis 2 years earlier. Pure tone audiometry, auditory brainstem response tests, and promontory tests did not reveal any cochlear activity. Computed tomography showed bilateral and totally ossified cochleae. Magnetic resonance imaging confirmed the diagnosis by showing no cochlear signal on T2-weighed images and ruled out brainstem and cerebellopontine angle abnormalities. INTERVENTION: A left auditory brainstem implantation was performed through a translabyrinthine route, using a Nucleus 22 channel device. MAIN OUTCOME MEASURES: Word and sentence recognition tests in sound-only and sound plus vision modes. RESULTS: No postoperative complication was observed. Twelve electrodes could be activated, and their tonotopy was defined. Nine other electrodes were inactivated because of an absence of auditory response (4 electrodes) or paresthesia (5 electrodes). At the last follow-up visit, 26 months after the implantation, 50% of disyllabic word scores and 60% of sentence scores were achieved using auditory brainstem implant sound only. These scores reached 80% and 93%, respectively, with lip-reading. CONCLUSION: Auditory brainstem implantation is an efficient means of auditory rehabilitation in cases of bilateral total hearing loss with totally ossified cochleae. It should be considered in cases of predictable failure in cochlear implantation. PMID- 12544034 TI - Recurrent extrusion of cochlear implant at an interval of 5 years. AB - OBJECTIVE: To describe two different types of device extrusion occurring in the same patient 1 and 6 years after cochlear implantation and to discuss the relation between the two extrusions and problems of cochlear implantation in patients with chronic suppurative otitis media. STUDY DESIGN: Retrospective case review. SETTING: Tertiary referral center. PATIENT AND INTERVENTIONS: The patient was a 52-year-old man with profound hearing loss in the right ear caused by chronic otitis media and deafness in the left ear caused by removal of an acoustic neuroma. Surgery for cleaning the tympanic cavity and reconstruction of the bony canal after previous radical mastoidectomy was performed in the right ear first. One year later, cochlear implantation was performed in this ear. RESULTS: One year after implantation, the anterior edge of the internal device of cochlear implant was exposed along the skin incision. This site was stable after corrective surgery. Five years after this operation, however, the posterior edge of the internal device extruded, accompanied by infection around the device. CONCLUSION: The two extrusions occurred with a long interval in between and at two different sites. The first extrusion occurred along the skin incision, and the second extrusion occurred away from the skin incision with infection around the device. The authors assumed that the first and the second extrusion occurred independently; the first extrusion was caused by insufficient blood supply resulting from scar formation from repeated mastoid operations and by pressure on the skin from the internal and external devices, and the second extrusion was caused by proliferated tissue pushing the device upward and against the skin as a result of chronic middle ear infection. The clinical course indicates that sufficiently long follow-up periods are needed to determine the usefulness of the prevention techniques for cochlear implantation in patients with chronic suppurative otitis media. PMID- 12544035 TI - The role of zinc in the treatment of tinnitus. AB - OBJECTIVE: This study was designed to investigate the role of zinc administration in treatment of tinnitus. STUDY DESIGN: Randomized, prospective, placebo controlled study. SETTING: Patients with tinnitus were admitted to the ear, nose, and throat clinic of the authors' hospital. PATIENTS: Patients with tinnitus with no know pathologic conditions of the ear, nose, and throat; the mean age of 28 patients receiving zinc was 51.2 years, and that of 13 patients given placebo was 55 years. INTERVENTION: Blood zinc levels were measured. Frequency was detected by audiometry, and loudness of tinnitus was screened by tinnitus match test. A questionnaire that scored tinnitus subjectively between 0 and 7 was given to patients before zinc treatment. After 2 months of treatment (zinc 50 mg daily to zinc group, placebo pill containing starch to placebo group), all of the tests were performed again. There was no difference in age, sex, duration of tinnitus, and affected ears between the patients treated with zinc and those treated with placebo. Blood zinc levels were lower than normal in 31% of patients before treatment. MAIN OUTCOME MEASURES: A decrease in tinnitus loudness by at least 10 dB was accepted as clinically favorable progress. A decrease of more than 1 point in subjective tinnitus scoring was accepted as valid. RESULTS: Clinically favorable progress was detected in 46.4% of patients given zinc. Although this decrease was not statistically significant, the severity of subjective tinnitus decreased in 82% of the patients receiving zinc. The mean of subjective tinnitus decreased from 5.25 +/- 1.08 to 2.82 +/- 1.81 ( < 0.001). However, the decrease in severity of the tinnitus was not significant in patients receiving placebo. CONCLUSION: It can be concluded that patients with tinnitus may have low blood zinc levels (31%) and clinical and subjective improvement can be achieved by oral zinc medication. However, it remains to be seen whether the longer duration of treatment has more significant results. PMID- 12544036 TI - Superficial siderosis of the meninges and its otolaryngologic connection: a series of five patients. AB - OBJECTIVE: To study the otolaryngologic disorders in patients with superficial siderosis of the meninges. BACKGROUND: Superficial siderosis of the central nervous system is a rare disorder characterized by progressive bilateral hearing loss and ataxia caused by recurrent bleeding into the subarachnoid space. The cerebellum, eighth cranial nerve, and olfactory nerve are particularly susceptible to the deposition of hemosiderin, which is responsible for the symptoms. The diagnosis is confirmed by magnetic resonance imaging. METHODS: The clinical notes of five patients with superficial siderosis of the meninges were reviewed with the intent of reporting the otolaryngologic symptoms and signs, the clinical investigations, and treatments. RESULTS: Four of the five patients had sensorineural deafness, two had smell disturbances, and three had gait abnormalities. Magnetic resonance imaging was the most important investigation used to identify the condition. CONCLUSIONS: Superficial siderosis of the meninges is an important differential diagnosis in patients with progressive sensorineural deafness. PMID- 12544037 TI - A new theory to explain the genesis of petrous apex cholesterol granuloma. AB - OBJECTIVE: To propose a new hypothesis that attempts to explain the pathogenesis of petrous apex cholesterol granuloma (PA CG). CLASSIC OBSTRUCTION-VACUUM HYPOTHESIS: PA CGs form when mucosal swelling blocks the circuitous pneumatic pathways to the apical air cells. Trapped gas resorption results in a vacuum that triggers bleeding, and CG forms through anaerobic breakdown of blood products. PROBLEMS WITH THE CLASSIC (OBSTRUCTION-VACUUM) HYPOTHESIS: Impaired ventilation of mucosa-lined pneumatic tracts in the middle ear, mastoid, paranasal sinuses, and lung are very common, but CG is rare. The extraordinary levels of temporal bone pneumatization typically observed in PA CG cases is indicative of excellent ventilation and freedom from inflammatory mucosal disease. Were under pressure due to gas absorption alone sufficient to trigger hemorrhage, CG ought to be frequent in otitis media with effusion. PATIENTS: The opposite PA of 13 patients with PA CG compared with 31 highly pneumatic PAs in patients undergoing imagery for non-otologic reasons. MAIN OUTCOME MEASURE: The nature of the bony partition, as seen on computed tomography, between the PA air cell system and the adjacent marrow compartment. RESULTS: 4 of 13 PAs with CGs on the opposite side showed deficient septation between air cells and marrow, whereas this was not observed in any of the 31 extensively pneumatized normal ears. NEW HYPOTHESIS (EXPOSED MARROW): As cellular tracts penetrate the apex during young adulthood, budding mucosa invades and replaces hematopoietic marrow. The bony interface becomes deficient, with coaptation of richly vascular marrow and the mucosal air cell lining. Hemorrhage from the exposed marrow coagulates within the mucosal cells and occludes outflow pathways. Sustained hemorrhage from exposed marrow elements provides the engine responsible for the progressive cyst expansion. As the cyst expands, bone erosion increases the surface area of exposed marrow along the cyst wall. This exposed marrow theory explains the unique proclivity of the healthy and well-pneumatized PA to form a CG. PMID- 12544038 TI - Cerebrospinal fluid leak after acoustic neuroma surgery: a comparison of the translabyrinthine, middle fossa, and retrosigmoid approaches. AB - OBJECTIVE: To determine whether the choice of surgical approach affects the rate of postoperative cerebrospinal fluid leakage in patients who have undergone surgical resection of acoustic neuroma. STUDY DESIGN: Retrospective chart review. SETTING: Tertiary referral center. PATIENTS: Three hundred patients who underwent surgery for acoustic neuromas were selected by consecutive medical record number until 100 resections via each surgical approach (translabyrinthine, middle fossa, and retrosigmoid) had been gathered. MAIN OUTCOME MEASURES: Surgical approach used, cerebrospinal fluid leak incidence, tumor size, patient age. RESULTS: Postoperative cerebrospinal fluid leak of any severity was observed in 13% of translabyrinthine, 10% of middle fossa, and 10% of retrosigmoid patients. These difference in the rate of cerebrospinal fluid leakage were not statistically significant (p = 0.82). The majority of leaks were managed conservatively with fluid and activity restriction, often accompanied by a period of lumbar subarachnoid drainage. There was a need to return to the operating room for a definitive procedure in 4% of translabyrinthine, 2% of middle fossa, and 3% retrosigmoid patients; again not statistically different among the approaches (p = 0.43). Tumor size was not correlated with cerebrospinal fluid leak rate (p = 0.13). Patient age, for patients older than 50 years, was suggestive of increased odds of cerebrospinal fluid leak (p = 0.06). CONCLUSION: Neither surgical approach nor tumor size affects the rate of postoperative cerebrospinal fluid leakage or the necessity of managing a leak with a return to the operating room. Cerebrospinal fluid leakage rates have remained stable in recent decades despite numerous innovative attempts to improve dural closure, seal transected air cell tracts, and occlude anatomic pathways. The finding that leak rates were similar among three dissimilar surgical techniques suggests that factors other than techniques of wound closure, such as transient postoperative rises in cerebrospinal fluid pressure, may be responsible for these recalcitrant cases. PMID- 12544039 TI - Acute otitis media and facial nerve paralysis in adults. AB - OBJECTIVE: The pathophysiology and treatment of facial nerve paralysis associated with acute otitis media are still under debate. The objective of this study was to review treatment strategies and extent of recovery in adult patients with the aim of defining a standard treatment protocol for this rare pathologic condition. STUDY DESIGN: Retrospective chart review. SETTING: University hospital, tertiary referral center. PATIENTS: Between 1993 and 2000, 11 patients were admitted for facial nerve paralysis secondary to acute otitis media. There were six women and five men without a history of chronic middle ear disease, who ranged in age from 21 to 71 years. Facial palsy was graded with the House-Brackmann scale: four patients had Grade III palsy, six had Grade IV palsy, and one patient had Grade V palsy. Bacteriologic examination of middle ear fluid was performed in four patients Streptococcus pneumoniae was observed in one patient, and the remaining three cultures were negative. INTERVENTIONS: All patients were treated with parenteral ampicillin-sulbactam or a third-generation cephalosporin in conjunction with oral or intravenous corticosteroids, except in a single patient with diabetes mellitus who received antibiotics alone. Myringotomy alone or with ventilation tube application was performed in eight patients. A simple mastoidectomy without facial nerve decompression was used in a patient with sudden impairment to Grade VI paralysis and worsening otitis after an initial improvement. RESULTS: Normal facial function returned in all patients, independently of the grade of the paralysis, the treatment strategy, or the outcome of the middle ear disease. The time of recovery varied from 2 weeks to 3 months, except for one patient who underwent mastoidectomy and in whom normal function returned in 10 months. CONCLUSIONS: The treatment of facial nerve paralysis secondary to otitis media should be as conservative as possible, using antibiotics and corticosteroids. Myringotomy and a ventilation tube should be added when spontaneous perforation of the tympanic membrane is not present. Mastoidectomy should be performed only when it is necessary to treat otitis media. Facial nerve decompression should not be necessary. PMID- 12544041 TI - Brainstem cavernous angioma. PMID- 12544040 TI - Significance of House-Brackmann facial nerve grading global score in the setting of differential facial nerve function. AB - OBJECTIVE: To determine the clinical significance of the House-Brackmann facial nerve grading scale (HBFNGS) in the setting of differential function along the branches of the facial nerve. STUDY DESIGN: Prospective study of 38 patients with facial palsy who demonstrated differential facial function. SETTING: Tertiary referral center. PATIENTS: Patients with facial nerve dysfunction from any cause. Patients with complete facial nerve paralysis (House-Brackmann Grade 6) were excluded. INTERVENTION: Physicians were provided with printed description of the HBFNGS and asked to report facial nerve function as a traditional global score and as a regional score based on the House-Brackmann scale for the forehead, eye, nose, and mouth. This was reported as F(w) E(x) N(y) M(z), where w, x, y, and z ranged from 1 to 6 based on the HBFNGS. Synkinesis was graded as none, mild, or severe. MAIN OUTCOME MEASURES: The traditional HBFNGS score was compared with a regional grading facial nerve grading system based on the HBFNGS for the forehead, eye, nose, and mouth. Agreement between the traditional global score and the regional scores was analyzed. RESULTS: In patients with variable facial weakness, the single House-Brackmann score did not fully communicate their facial function. Further, the single grade did not always correlate with the best or worst function along the four facial regions. The single House-Brackmann score most strongly correlated with the regional scoring of the eye (61%), followed by the nose/midface (40%), mouth (32%), and forehead (18%). The global score did not correlate with the worst regional score in 30 patients (79%). In 3 of 5 patients with synkinesis and an obligatory Grade 3 or higher in the global House-Brackmann grading system, the regional facial function was Grade 2 or better at one or more areas of the face. CONCLUSIONS: In patients with differential facial function, a single global number is inadequate to describe facial function and primarily reflects the function of the eye. Regional assessment using the HBFNGS and reported as F(w) E(x) N(y) M(z) more fully communicates facial function. PMID- 12544042 TI - Scheibe dysplasia. PMID- 12544044 TI - Addressing the nursing shortage: our voices are also being heard outside of nursing. PMID- 12544045 TI - Future trends in nursing employment: the nursing shortage has made choosing the right career path easier. PMID- 12544046 TI - Making informed employment decisions: questions to ask and issues to consider. PMID- 12544047 TI - Charting the course of your nursing career: finding the joy in being a nurse. PMID- 12544048 TI - Results of a multistate licensure survey. PMID- 12544049 TI - How to get a license. PMID- 12544050 TI - Your guide to certification: earning additional credentials can significantly enhance your career. Here's what you need to know to pursue them. PMID- 12544051 TI - Honoring nursing's philanthropists. PMID- 12544052 TI - Shocking Edith. PMID- 12544054 TI - Debate intensifies over health care issues. PMID- 12544056 TI - The left ventricular assist device. AB - Of the 400,000 people in the United States who develop end-stage heart failure each year, 60,000 are unresponsive to medical therapy and 2,500 undergo heart transplantation. Surgically implanted pumps, called LVADs, are extending many lives. PMID- 12544057 TI - Failure to rescue. PMID- 12544058 TI - Ethical concerns in end-of-life care. PMID- 12544062 TI - Media relations 101. PMID- 12544063 TI - Screening for osteoporosis in postmenopausal women: recommendations and rationale. PMID- 12544064 TI - Sexuality and chronic pain. PMID- 12544065 TI - Nurse anesthetists: one step forward, one step back. PMID- 12544066 TI - Nursing care in assisted living facilities. PMID- 12544067 TI - Rose petal corridors: homicide at a college of nursing. PMID- 12544068 TI - Smallpox vaccines. PMID- 12544069 TI - Resistance to hoop stress in balloon expandable stents: evaluation in an ex vivo model. AB - RATIONALE AND OBJECTIVES: To develop a new model for a more realistic evaluation of radial strength and elastic recoil of balloon expandable stents using a new ex vivo model in human cadaver bifurcations of the aorta. MATERIALS AND METHODS: Four different stents (each group n = 10) were implanted in cadaver common iliac arteries. Randomization was performed either to right or left iliac artery. The specimens were cast filled with silicone caoutchouc and after 24 hours the vascular walls including the stents were removed from the hardened casts. The weight of the cast cylinders of the stents was measured in air and in purified water: the difference of the two values resulted in the buoyancy force and because of that the volume of the bodies as a relative degree for the radial strength could be calculated. The findings were correlated with the workbench tests of the manufacturers. RESULTS: Manufacturer's workbench tests were incomparable because of wide spread specifications. There was a significant difference between the theoretical maximal volume (0.866 mL) and the real cast volumes ( P< 0.025) that corresponds directly to the elastic recoil of the stents. The following mean real cast volumes were measured (corrected for 1 cm): Sinus-Stent 0.677 mL, Palmaz-Stent 0.708 mL, Jostent 0.715 mL, Saxx-Stent 0.732 mL, thus reflecting the various degree of radial strength; therefore, the ranking in radial strength resistance was Saxx, Jostent, Palmaz, and Sinus. CONCLUSION: High radial strength and low elastic recoil are important requirements of any stent design. Ex vivo tests unlike in vitro physical testing facilitate a realistic evaluation of the inherent stent characteristics. The model used in this study proved to be uniformly valid for physical stent design testing. PMID- 12544070 TI - Visualization of breast lymphatic pathways with an indirect computed tomography lymphography using a nonionic monometric contrast medium iopamidol: preliminary results. AB - RATIONALE AND OBJECTIVES: The capability of an indirect computed tomographic lymphography (CT-LG) using a nonionic monometric contrast medium iopamidol for visualizing breast lymphatic pathways was preliminarily tested. MATERIALS AND METHODS: In 10 female dogs, a total of 0.5 and 1 mL of undiluted iopamidol was injected subcutaneously into the skin areas overlying the both caudal mammary glands. Contiguous 2-mm-thick multidetector raw helical CT images were obtained through the upper thorax and axilla before and during 60 minutes after gentle massage at the injection sites, with reconstruction into three-dimensional (3D) postcontrast CT images. The first lymph node (1st LN) directly draining from the injection sites was marked under CT guidance, followed by pre- and postmortem examinations. This CT-LG with 2-mL iopamidol was also attempted in five human female volunteers. RESULTS: Even with 0.5-mL iopamidol, the CT-LG clearly visualized the direct connection of the 1st LN and lymphatic vessels draining from the injection sites throughout the examination time in all the animals, with the maximum CT attenuation of 269 Hounsfield units (HU) +/- 137 in the 1st LN on the first postcontrast images. The topographic 3D images provided comprehensive anatomic outlines of these lymphatic pathways. Of the total of 20 opacified 1st LN and 110 distant nodes, all the 1st LN (100%) and 92 (83.6%) distant nodes could be resected at pre- or postmortem, with a good correlation with the CT images. The CT-LG also effectively localized the 1st LN with the maximum attenuation of 223 HU +/- 63 in the human volunteers, without any significant late adverse effects. CONCLUSION: Indirect CT-LG with iopamidol may have excellent potential for visualizing breast lymphatic drainage and for preoperative localization of breast sentinel lymph nodes. PMID- 12544071 TI - Low-dose gadobenate dimeglumine versus standard dose gadopentetate dimeglumine for contrast-enhanced magnetic resonance imaging of the liver: an intra individual crossover comparison. AB - RATIONALE AND OBJECTIVES: Gadobenate dimeglumine (Gd-BOPTA) has a two-fold higher T1 relaxivity compared with gadopentetate dimeglumine (Gd-DTPA) and can be used for both dynamic and delayed liver MRI. This intraindividual, crossover study was conducted to compare 0.05 mmol/kg Gd-BOPTA with 0.1 mmol/kg Gd-DTPA for liver MRI. MATERIALS AND METHODS: Forty-one patients underwent two identical MR examinations separated by >or= 72 hours. Precontrast T1-FLASH-2D and T2-TSE sequences and postcontrast T1-FLASH-2D sequences were acquired during the dynamic and delayed (1-2 hours) phases after each contrast injection. Images were evaluated on-site by two independent, blinded off-site readers in terms of confidence for lesion detection, lesion number, character and diagnosis, enhancement pattern, lesion-to-liver contrast, and benefit of dynamic and delayed scans. Additional on-site evaluation was performed of the overall diagnostic value of each agent. RESULTS: Superior diagnostic confidence was noted by on-site investigators and off-site assessors 1 and 2 for 6, 4 and 2 patients with Gd BOPTA, and for 3, 1 and 2 patients with Gd-DTPA, respectively. No consistent differences were noted for other parameters on dynamic phase images whereas greater lesion-to-liver contrast was noted for more patients on delayed images after Gd-BOPTA. More correct diagnoses of histologically confirmed lesions (n = 26) were made with the complete Gd-BOPTA image set than with the complete Gd-DTPA set (reader 1: 68% vs. 59%; reader 2: 78% vs. 68%). The overall diagnostic value was considered superior after Gd-BOPTA in seven patients and after Gd-DTPA in one patient. CONCLUSION: The additional diagnostic information on delayed imaging, combined with the possibility to use a lower overall dose to obtain similar diagnostic information on dynamic imaging, offers a distinct clinical advantage for Gd-BOPTA for liver MRI. PMID- 12544072 TI - Vascular responses in normal canine carotid arteries: comparison between various self-expanding stents of the same unconstrained size. AB - RATIONALE AND OBJECTIVES: To compare long-term vascular responses upon the insertion of various self-expandable stents, all the same unconstrained size, in canine carotid artery models. MATERIALS AND METHODS: Twenty-two stents (5 SMARTs, 5 Wallstents, 6 Niti-Ss, 6 Niti-Ds) of the same unconstrained size (6 mm in diameter, 20 mm in length) were endovascularly placed in canine common carotid arteries. The luminal changes were measured on three occasions, on prestenting, immediate poststenting, and angiograms taken before specimens were killed. After en-bloc harvest of the stented carotid arteries at 6 months, the intraluminal surface was evaluated by gross observation and scanning electron microscopy (SEM). Neointimal thickness was measured at several points both over the wire and between the wires. RESULTS: Niti-D was excluded from analysis because of high rate of poststenting occlusion. SMART stent showed the greatest expansibility with average initial luminal gain ( < 0.05) of 21.2% (Niti-S: 16.5%, Wallstent: 12.9%). At 6 months follow-up, the dilated arterial lumen had returned almost to the prestenting caliber without any significant differences among the stent types ( > 0.05). The thickness of neointimal coverage was more prominent with SMART stent (354 microm over the wire and 258 microm between the wires) than Niti-S (228 microm and 83 microm) or Wallstent (187 microm and 78 microm). CONCLUSION: Stent types with its higher initial luminal gain appeared to be associated with thicker neointimal formation at 6 months. The acute expanding force of a self expanding stent may be the key to the cause of neointimal hyperplasia. Regardless of the inserted stent type, the variations in neointimal response were offset by luminal gains of varying degree, thus preserving the arterial patency almost to the prestenting size. PMID- 12544073 TI - Dynamic gadolinium uptake in thermally treated canine brain tissue and experimental cerebral tumors. AB - RATIONALE AND OBJECTIVES: Thermal coagulation of cerebral tumors induces reactive changes within adjacent brain tissue, which appear as Gd-DTPA enhancement in MR images. This makes assessment of therapeutic success difficult to establish radiographically because the reactive changes can mimic residual tumor. Dynamic Gd-DTPA uptake curves in reactive tissue and tumor were investigated to assess the utility of contrast enhanced (CE)-dynamic MRI to distinguish reactive changes from residual tumor in a canine model. MATERIALS AND METHODS: Cerebral thermal necrosis was induced using a 980 nm laser in 11 dogs with intracerebral transmissible venereal tumors (TVTs). A fast spin-echo T1-weighted imaging sequence was used for CE-dynamic MRI. Gd-DTPA uptake data were acquired with 10 second temporal resolution and for untreated TVTs for reactive tissue using a sigmoidal-exponential model. RESULTS: Characteristic gadolinium uptake curves were measured and characterized for reactive brain tissue, and untreated and treated TVTs. Both early and delayed dynamic responses were significantly different in reactive brain tissue compared with TVT. CONCLUSION: Reactive thermal changes in otherwise normal brain tissue can be distinguished from residual tumor after cerebral thermal therapy using CE-dynamic MRI. PMID- 12544074 TI - Evaluation of reconstruction windows for multislice computed tomography in quantification of coronary calcium. AB - RATIONALE AND OBJECTIVES: To search for an optimum reconstruction window in retrospectively gated multislice computed tomography (MSCT) for quantification of coronary calcium. MATERIALS AND METHODS: Coronary calcium quantified was examined as Agatston and volume scores by two experienced observers at 10 time points across the R-R interval of the electrocardiogram in 42 patients. A combination of statistical approaches was used to evaluate the distributions of minimum and maximum scores and of interobserver variability for both scoring methods across the cardiac cycle. RESULTS: Based on the combination of evaluation approaches, 60% to 70% of the R-R interval appeared to be the optimum time point for obtaining maximum calcium scores with minimum interobserver variability. The optimum time point was more clearly defined for the Agatston score than for the volume score. CONCLUSION: A reconstruction window beginning at 60% to 70% of the R-R interval seems to be most advantageous for retrospective gating of MSCT studies performed to quantify coronary calcium. PMID- 12544075 TI - Comparison of image quality in contrast-enhanced coronary-artery visualization by electron beam tomography and retrospectively electrocardiogram-gated multislice spiral computed tomography. AB - RATIONALE AND OBJECTIVES: To compare the image quality of electron beam tomography (EBT) and multislice spiral CT (MSCT) for coronary artery visualization. MATERIALS AND METHODS: Two groups of 30 patients without coronary stenoses were studied by MSCT (4 x 1 mm collimation) or EBT (3 mm slice thickness). Contrast-to-noise ratio (CNR), overall length of the visualized arteries and vessel length free of motion artifacts were measured. RESULTS: Length of visualized arteries was equal in MSCT and EBT. In EBT, longer segments were depicted free of motion artifacts (MSCT: 73%, EBT: 92% of visualized length, P< 0.001) and CNR was significantly higher than in MSCT (15.4 vs. 9.0; P< 0.001). In both modalities, vessel diameters correlated closely to quantitative coronary angiography. CONCLUSIONS: EBT and MSCT permit reliable coronary artery visualization and measurement of vessel diameters. For the used scan protocol, MSCT images had a lower CNR and were more frequently affected by motion. PMID- 12544076 TI - Percutaneous radiofrequency thermal ablation of lung VX2 tumors in a rabbit model using a cooled tip-electrode: feasibility, safety, and effectiveness. AB - RATIONALE AND OBJECTIVE: To evaluate the feasibility, safety, and effectiveness of percutaneous CT-guided transthoracic radiofrequency ablation (RFA) of lung VX2 tumors implanted in rabbits. MATERIALS AND METHODS: Thirty-five rabbits with implanted lung VX2 tumors were divided into two groups, an RFA group (n = 28) and a control group (n = 7). In the RFA group, RFA was performed on VX2 tumors using a 17-gauge internally cooled-tip electrode. Contrast-enhanced CT was performed before the RFA and on the same day, day 3, weeks 1, 2, 3, and months 1, 2, 6, and 9, after the RFA. The therapeutic efficacy was evaluated by CT and pathologic findings. RESULTS: RFA of lung tumors was technically successful in each instance. Complete tumor ablation was achieved in 19 of the 28 rabbits (67.9%) in the RFA group. Nine rabbits (32.1%) showed local tumor relapse and mediastinal lymph nodal or pleural metastasis. The animals in the control group died of end stage malignancies with diffuse tumor spread and malignant pleural effusion (mean 26 +/- 2.7 days). There were 17 (60.7%) complications related to the procedure, ie, pneumothorax (n = 12), obstructive pneumonia (n = 3), hemothorax (n = 1), and burn (n = 1). RFA of centrally located VX2 tumors (in inner 2/3 of the lung) was more frequently associated with complications than RFA of peripherally located VX2 tumors ( P= 0.02). CONCLUSION: This experimental study demonstrates the feasibility of RFA therapy for treating lung VX2 tumors in rabbits, although RFA for central tumors carries the potential for major complications, including large pneumothorax or obstructive pneumonia. PMID- 12544077 TI - Glucocorticoid-induced osteoporosis in patients with sarcoidosis. AB - BACKGROUND: Patients with sarcoidosis are at risk for osteoporosis caused by glucocorticoid therapy. However, because of potential hypercalciuria and hypercalcemia, the usual conservative treatment for low bone mass, calcium and vitamin D supplements, may not be well tolerated. METHODS: Patients with sarcoidosis referred to a metabolic bone clinic were compared with other sarcoidosis patients studied prospectively and patients with chronic obstructive pulmonary disease (COPD) or asthma. The subjects underwent bone mineral density (BMD) testing, and the sarcoidosis patients underwent mobility testing and measurements of serum and urine chemistries, vitamin D levels, bone turnover markers, and sex hormone levels. RESULTS: The subjects were mostly male African Americans in the 6th decade of life. Many took chronic oral glucocorticoid therapy and often used home oxygen therapy. Low hip BMD was common among the referred group, comparable with patients with COPD. Surprisingly, hypercalciuria and hypercalcemia were uncommon, and serum testosterone levels were frequently low. The use of calcium supplements, multivitamins containing vitamin D, and glucocorticoids had no impact on serum or urine calcium levels. From univariate analysis, potential risk factors for low hip BMD were low weight, low body mass index (BMI), advanced age, and current use of glucocorticoids. However, in stepwise multiple regression analysis, only low BMI predicted about 40% of hip BMD. CONCLUSIONS: Despite calcium and vitamin D supplements, this group of patients with sarcoidosis had low BMD but relatively infrequent hypercalciuria and hypercalcemia. No prediction model of BMD was adequate. Therefore, we conclude that each patient needs to be assessed individually, including measurement of BMD, serum and urine calcium, and sex steroid status. PMID- 12544078 TI - Evaluation and treatment of acute bronchitis at an academic teaching clinic. AB - BACKGROUND: Randomized controlled trials have demonstrated that antibiotics provide no benefit for acute bronchitis, yet 55 to 90% of patients who receive this diagnosis are treated with antibiotics. Given substantial data against antibiotics for acute bronchitis, it could be expected that physicians at academic teaching institutions would be less likely to prescribe antibiotics. However, limited data of antibiotic use for acute bronchitis in this setting has been published. METHODS: Charts of patients seen between January 1 and October 25, 2000, who received an ICD-9 diagnosis of acute bronchitis or upper respiratory infection (URI) at the University of Virginia internal medicine clinic were reviewed. Patients were excluded if they had no cough, chronic obstructive pulmonary disease, symptoms for > or = 3 weeks, or antibiotics for another reason. RESULTS: Of the 160 patients included in this study, 105 (66%) received an antibiotic. Multivariate analysis revealed that patients with increasing age (P = 0.002), purulent cough (P = 0.003), abnormal exam (P = 0.003), and comorbidities (P = 0.03) were most likely to receive an antibiotic. Smoking, duration of symptoms, gender, and race did not predict antibiotic use (P > 0.05). Macrolides accounted for 68% of antibiotics. Twenty-two (14%) of all patients received a chest radiograph and 72 (45%) received an inhaler. Of those who had chest radiographs negative for signs of infection, 76% received an antibiotic. CONCLUSION: In our teaching clinic, antibiotics were overused, whereas chest radiographs and inhalers were underused for the evaluation and treatment of acute bronchitis. Recently published guidelines will help curb use of antibiotics, but a more intensive intervention, including physician and patient education is probably necessary. PMID- 12544079 TI - Loss of laboratory instruction in American medical schools: erosion of Flexner's view of "scientific medical education.". AB - There has been a steep decline in the number of hours devoted to laboratory instruction in American medical schools. Medical students attending 1 of the 3 Washington DC medical schools now spend less than 10% of their first 2 basic science years in a laboratory. The paucity of laboratory instruction represents a reversal of the gains made in American medical education after the Flexner report and may partly account for our nation's missing physician-scientists. This situation is not expected to improve anytime soon, given the expenses that would be required to divert research-intensive faculty to laboratory instruction. The expenses would be particularly onerous for medical schools under intensive managed care pressures. Because it is unlikely that many American medical schools have either the will or means to make substantive changes in their laboratory based curricula, novel solutions to restoring laboratory-based medical education may be required. PMID- 12544080 TI - Cardiology Grand Rounds from the University of North Carolina at Chapel Hill. The antioxidant vitamins and coronary heart disease: Part II. Randomized clinical trials. PMID- 12544081 TI - Southwestern Internal Medicine Conference. Etiology and management of delirium. AB - Delirium has been recognized for the last 3 millennia and is the most common complication found in hospitalized patients aged 65 and older in the United States. However, critical basic science and clinical research did not progress until the DSM III criteria clearly defined delirium 20 years ago. The term delirium then replaced many nonspecific entities, such as acute confusion state, acute brain syndrome, metabolic encephalopathy, and toxic psychosis. This review discusses the epidemiology, risk factors, interventions, causes, management, and outcomes of delirium. The pathophysiology of delirium has the potential to radically alter our management of delirium and is a controversial area of research. PMID- 12544082 TI - Hepatitis associated with amoxicillin/clavulanic acid and/or ciprofloxacin. AB - We describe an elderly patient with normal pre-existing liver functions who was treated with amoxicillin/clavulanic acid and later ciprofloxacin for acute bronchitis. He developed a pattern of liver dysfunction consistent with hepatocellular injury, with clinical features of a hypersensitivity reaction, which may be attributable to either or both of the antimicrobial agents used. This gradually resolved over a 4-week time period, with conservative management. A review of the relevant literature on drug-induced hepatotoxicity is also presented. PMID- 12544083 TI - Kikuchi disease presenting as a flu-like illness with rash and lymphadenopathy. AB - Kikuchi-Fujimoto disease (Kikuchi Disease) is a self-limited and benign systemic lymphadenitis of unknown cause, originally described by Kikuchi and Fujimoto and coworkers in 1972. Although relatively uncommon, it is increasingly discussed in the medical literature. Clinical presentation typically includes adenopathy, particularly cervical, with fever and flu-like symptoms. This constellation of symptoms, in the presence of a characteristic histiocytic necrotizing lymphadenitis, provides the clinicopathologic diagnosis. The immunopathogenesis of Kikuchi disease may lie in a hyperactive response to viral infection. We describe an African American man with Kikuchi disease, unusual in the extent of his rash and debilitation, and in the relapse of his clinical symptoms. PMID- 12544084 TI - Spontaneous tumor lysis syndrome in solid tumors: really a rare condition? AB - Acute tumor lysis syndrome (ATLS), which occurs spontaneously, without cytotoxic therapy, is a rare condition. Spontaneous TLS (STLS) has been seen most commonly in lymphoma and leukemia. We report a series of 3 cases of STLS in patients with solid tumors who were hospitalized in our department during a 9-month period and suggest that STLS is probably more frequent than previously thought. PMID- 12544085 TI - Primary antiphospholipid syndrome associated with acute adrenal failure. AB - We describe a 48-year-old woman with primary antiphospholipid syndrome who developed acute adrenal failure after an operation for a uterine myoma. After surgery, she developed a preshock state with hypotension, hypoglycemia, and hyponatremia. A diagnosis of primary antiphospholipid syndrome was made based not only on her past history of skin ulceration and recurrent spontaneous abortions but also on the presence of anticardiolipin antibodies. An abdominal computed tomography showed a bilateral enlargement of the adrenal glands but no high density region in either gland. The patient recovered from the shock-like syndromes after the administration of glucocorticoids. Because it is possible that patients with antiphospholipid syndrome have acute or chronic adrenal failure caused by repeated hemorrhage or thrombosis, it may be important to monitor adrenal function in patients when the presence of this antibody is detected. PMID- 12544086 TI - Heparin-induced thrombocytopenia with pulmonary embolism and disseminated intravascular coagulation associated with low-molecular-weight heparin. AB - Heparin-induced thrombocytopenia (HIT) is a severe adverse effect of heparin therapy. Although most cases occur in patients receiving unfractionated heparin, HIT can arise in venous thrombosis prophylaxis with a low-molecular-weight heparin (LMWH). We report an uncommon case of HIT in a postoperative orthopedic patient associated with LMWH (nadroparin), complicated by deep venous thrombosis, pulmonary embolism, and disseminated intravascular coagulation, treated successfully with recombinant hirudin and immunoglobulin therapy. PMID- 12544087 TI - Thyrotropin-secreting pituitary adenoma presenting as hypokalemic periodic paralysis. AB - Thyrotropin (TSH)-secreting pituitary adenoma presenting with hypokalemic periodic paralysis is extraordinarily rare and may be misdiagnosed. We describe a 44-year-old man who suffered from acute muscle weakness and inability to ambulate upon awakening in the morning. Physical examination showed hypertension, tachycardia, and symmetrical flaccid paralysis of all extremities. The major biochemical abnormality was hypokalemia (K+, 2.0 mmol/L) with low urine K+ excretion. A thyroid function study revealed elevated thyroid hormone levels and inappropriately high TSH concentrations (2.10 microU/mL). Brain magnetic resonance imaging delineated a pituitary tumor with suprasellar extension. After trans-sphenoidal removal of tumor, he became clinically and biochemically euthyroid without any further attack of paralysis. Pathological findings confirmed a TSH-secreting adenoma with exclusive TSH immunostaining. TSH secreting pituitary adenoma must be kept in the differential diagnosis in any thyrotoxic periodic paralysis patients with detectable TSH levels to avoid delaying diagnosis and management. PMID- 12544088 TI - Pericardial tamponade-associated hyponatremia. AB - We describe a 58-year-old patient with adenocarcinoma of the lung who suffered from severe weariness that was attributed to hyponatremia. The cause of hyponatremia was found to be a malignant pericardial effusion that caused pericardial tamponade. The hyponatremia was corrected rapidly after the evacuation of the malignant effusion. The pathogenetic mechanisms of hyponatremia secondary to pericardial effusion are discussed. PMID- 12544091 TI - Pigmented atypical fibroxanthoma: a tumor that may be easily mistaken for malignant melanoma. AB - Nine cases of an unusual presentation of atypical fibroxanthoma that we previously termed pigmented atypical fibroxanthoma are reported. Pigmented atypical fibroxanthoma can be easily mistaken for malignant melanoma both clinically and histopathologically. The resemblance of pigmented atypical fibroxanthoma to melanoma is secondary to the ability of neoplastic cells to ingest and degrade erythrocytes following intratumoral hemorrhage and to accumulate hemosiderin in their cytoplasm. The histopathologic diagnosis of pigmented atypical fibroxanthoma can be easily confirmed by immunohistochemistry and iron stain. PMID- 12544092 TI - Gene expression profiling of melanocytic lesions. AB - DNA microarrays, microscopic grids of DNA, can be used to assess gene expression within a particular cell or cell population. Since DNA from thousands of genes can be hybridized and analyzed in one experiment, researchers can globally characterize genes expressed in normal and various pathologic states. To accurately assess the differences between normal and pathologic states, one derives cDNA from control and diseased tissue specimens for genes expression profiling. For these reasons, microarray technology may be of particular interest to dermatopathologists and dermatologists interested in understanding cutaneous disease because these physicians have access to tissue specimens. In addition, microarray technology is an efficient way of identifying molecules expressed in a cell population; therefore, it can be used to search for unique immunohistologic markers. To this end, we have used microarray technology to define differences in the gene expression profile of nevi and melanomas. In this manuscript, we discuss the results of our study, which confirm previously known differences in gene expression between melanoma and nevi. While a few genes appear slightly overexpressed in nevi, a number of genes involved in regulating cell proliferation were upregulated in melanoma, such as cyclin D1, cdc2-related protein kinase, c-Myc binding protein, early growth response protein 1, and pleiotrophin. "Housekeeping" genes such as glyceraldehyde 3-phosphate dehydrogenase were expressed at similar levels in melanoma and nevi. Surprisingly, a majority of genes were expressed at similar levels in both nevi and melanoma. Based on this study, DNA microarray technology appears to be a valuable tool for identifying genes that may be specifically expressed in cutaneous lesions. PMID- 12544093 TI - Oral melanoacanthoma: a report of 10 cases, review of the literature, and immunohistochemical analysis for HMB-45 reactivity. AB - Oral melanoacanthoma (MA) is rare reactive mucosal lesion that, like cutaneous MA, demonstrates hyperplasia of spinous keratinocytes and melanocytes. Unlike MA of the skin, oral MA is unrelated to seborrheic keratosis. This series adds 10 cases to the limited number of previous reports of oral MA. The clinicopathologic features of the cases in this series are generally consistent with those previously reported in the literature; that is, although documented in various intraoral locations in patients of differing ethnicity, oral melanoacanthoma most often presents as an enlarging flat or slightly raised area of hyperpigmentation on the buccal mucosa of adult black women. The current series provides evidence of occurrence over a wider age range (5-77 years) than previously reported. Additionally, the reactivity of oral melanoacanthoma to HMB-45 was investigated. Strong HMB-45 reactivity was present in all cases, thus demonstrating its limited utility in distinguishing oral MA from malignant melanoma. PMID- 12544094 TI - Ultrastructural studies in stable vitiligo. AB - Vitiligo is a disease of melanocytes characterized by achromic lesions in the skin, affecting the epidermis and the pilosebaceous follicle. We performed an ultrastructural analysis of biopsy specimens from four patients with noninflammatory, stable vitiligo of long duration (three had generalized vitiligo and one had segmental vitiligo). The samples were taken from the oldest achromic lesions, and the biopsy sites were far from normal skin. In all cases we noted alterations in keratinocytes, Langerhans cells, and melanocytes. We also found lymphocytes in the epidermis, and these cells and macrophages were noted in the dermis. The basal membrane disappeared at some points, and sometimes it was possible to see dermal cells with processes that engulfed either granular material or vesicles of epidermal origin in such areas. Our studies suggest that even in stable vitiligo, achromia implies intense cytologic activity, probably involving cell-mediated cytotoxicity, and ultrastructural findings resemble those of a lichenoid reaction. PMID- 12544095 TI - Malignant blue nevus: a case report and molecular analysis. AB - Malignant blue nevus is a rare melanocytic tumor that is described by some authors as a variant of malignant melanoma, whereas others regard it as a distinct entity. To our knowledge no molecular studies of this tumor have been performed, although the molecular pathogenesis of conventional melanomas has been extensively described. We present a case of malignant blue nevus that developed in a 15-cm congenital blue nevus on the back of a 41-year-old man. Subsequent regional lymph node and lung metastases developed within 1 and 29 months, respectively. We performed a molecular analysis for loss of heterozygosity on microdissected samples from the spectrum of benign to malignant blue nevus, using a panel of eight genes (MTS1, MXI1, CMM1, p53, NF1, L-myc hOGG1, and MCC), many of which are commonly associated with conventional melanomas. No loss of heterozygosity was detected, despite informativeness in seven genes. We suggest that malignant blue nevus may represent a distinct entity with a different molecular pathway to tumorigenesis than that of conventional melanomas. PMID- 12544096 TI - Herpesvirus 8-associated penile Kaposi's sarcoma in an HIV-negative patient: first report of a solitary lesion. AB - Kaposi's sarcoma is a neoplastic vascular lesion. Its form of onset is frequently disseminated, especially in HIV-positive patients. Its association with the infection caused by a virus of the Epstein-Barr family, human herpesvirus 8 (HHV 8), has been recently demonstrated. In this article we discuss the unusual presentation of a solitary manifestation of Kaposi's sarcoma on the penis of a 53 year-old HIV-negative patient. Polymerase chain reaction analysis of the tumor tissue was positive for HHV-8 in the tumor cells but not in the reactive stroma cells surrounding the tumor. The case is interesting for its unusual site of presentation, the young age of onset, the association with HHV-8 infection, the HIV-negative serology, and the benign course of the disease. PMID- 12544097 TI - Neuroblastoma-like schwannoma: a case report and review of the literature. AB - We report a case of schwannoma (neurilemmoma) predominantly composed of small cells arranged in rosettes around central collagenous cores and discuss the differential diagnosis of this unusual variant. Schwannoma with giant rosettes, previously designated as neuroblastoma-like schwannoma by Goldblum et al., must be differentiated from neuroblastoma, peripheral neuroectodermal tumors, and malignant change in a schwannoma. PMID- 12544098 TI - Rosette formation within a proliferative nodule of an atypical combined melanocytic nevus in an adult. AB - Rosette formation is a feature that has not been described as occurring in melanocytic neoplasms. We present such a unique case. A 59-year-old man presented with an asymptomatic, soft, hairy 3.0 x 2.0-cm pigmented lesion that had been present for many years in the right external ear, extending from the conchal bowl onto the antitragus area. Examination of histologic sections showed a proliferation of nonatypical and heavily pigmented melanocytes in the superficial dermis and around deep adnexal structures, characteristic of a congenital nevus. In other areas, pigmented spindled and dendritic cells infiltrated thickened collagen bundles in a pattern of a blue nevus. A nodular proliferation of epithelioid melanocytes was seen within the deep dermis and subcutaneous tissue. The periphery of the nodule merged with the surrounding nevus cells. Neoplastic cells with nuclear atypia, melanin pigment, pseudonuclear inclusions, and balloon cell change were present. In addition, there was rosette formation by the tumor cells, with a central aggregate of coarse cell processes. Neuroid cords were also noted. No prominent mitotic figures, necrosis, or significant inflammatory infiltrate were noted. The neoplastic cells were positive for S-100 protein, Mart 1, tyrosinase, neuron-specific enolase, and vimentin. HMB-45 and Ki-67 (MIB-1) labeled only rare neoplastic cells within the proliferative nodule. The tumor cells were negative for synaptophysin, protein gene product 9.5, CD57, epithelial membrane antigen, CD31, and CD34. The central cell processes of the rosettes were negative for trichome, type IV collagen, neurofilament protein, glial fibrillary acidic protein, and tyrosine hydroxylase. We also retrospectively examined 78 congenital nevi of 65 pediatric patients at our institution. Rosette formation was not seen in any of these cases. PMID- 12544099 TI - Ganglioneuroblastic differentiation in a primary cutaneous malignant melanoma. AB - Ganglioneuroblastic differentiation in malignant melanomas is an exceedingly rare event. Although there has been a single report of this occurrence in a metastatic melanoma, divergent ganglioneuroblastic differentiation has not been documented previously in a primary cutaneous lesion of melanoma. The present report describes an unusual case of invasive melanoma arising on the lower leg of a 61 year-old woman. The 16.9-mm thick tumor showed extensive ganglioneuroblastic differentiation, which was confirmed both immunohistochemically and ultrastructurally. Although the prognostic significance of this observation remains uncertain, the unique case reaffirms the potential morphologic diversity of melanomas and suggests a shared histogenetic origin from a common neural crest derivative. PMID- 12544100 TI - Embryonal rhabdomyosarcoma presenting in an adult: a case report and discussion of immunohistochemical staining. AB - Embryonal rhabdomyosarcoma is the most common soft tissue sarcoma of childhood but is rarely seen in adults. We report an embryonal rhabdomyosarcoma of the cheek in a 21-year-old Filipino man presenting as a rapidly enlarging mass. An incisional biopsy was consistent with embryonal rhabdomyosarcoma. The patient failed to respond to one cycle of chemotherapy after initial diagnosis and workup for metastatic disease. Wide local excision of the tumor was then completed with the use of the Mohs technique to clear the glabrous cutaneous margins during surgery. A Mohs surgeon and dermatopathologist were involved in interpretation of the Mohs sections. Subsequent immunohistochemical staining revealed a poorly differentiated spindled and epithelioid cell tumor, which stained diffusely positive for vimentin, S-100, and MyoD1. Rare rhabdomyoblasts were present and stained positively for desmin, muscle-specific actin, and phosphotungstic acid hematoxylin. The patient subsequently underwent radiation therapy for a total of 6,000 cGy, followed by two cycles of chemotherapy. The patient continues to be disease-free at 22 months after his wide local excision. PMID- 12544101 TI - Calcospherules associated with juvenile hyaline fibromatosis. AB - Juvenile hyaline fibromatosis is a rare autosomal recessive genodermatosis that manifests with 1 to 5 cm skin tumors, gingival hyperplasia, osteolytic bone lesions, and joint contractures. The skin tumors are most often located on the head and periarticular extremities. A case of juvenile hyaline fibromatosis with typical skin tumors that on biopsy exhibited basophilic calcospherules is presented. Although noted in a single case, these structures may be a novel, yet helpful, diagnostic marker. PMID- 12544102 TI - Granuloma annulare-like infiltrates with concomitant cutaneous involvement by B cell non-Hodgkin's lymphoma: report of a case. AB - Granulomatous infiltrates, including granuloma annulare-like changes, in patients with T-cell non-Hodgkin's lymphoma and Hodgkin's disease are well described but are exceedingly uncommon in B-cell non-Hodgkin's lymphoma. Herein, we describe a 73-year-old male with a 10-year history of B-cell chronic lymphocytic leukemia who developed erythematous annular plaques symmetrically on his upper extremities. Biopsies of these lesions revealed both granuloma annulare-like areas, in conjunction with nodular atypical lymphoid infiltrates consistent with secondary involvement by B-cell chronic lymphocytic leukemia. To our knowledge, this is the first report of cutaneous lesions resembling granuloma annulare with concomitant involvement by B-cell chronic lymphocytic leukemia. Awareness of this association may prevent nonrecognition of a neoplastic B-cell infiltrate in a granuloma annulare-like process. PMID- 12544103 TI - A case of lymphomatoid papulosis with prominent myxoid change resembling a mesenchymal neoplasm. AB - Lymphomatoid papulosis is a chronic recurrent eruption of self-regressing papules and nodules. Histologically this disorder is characterized by large atypical lymphoid cells resembling Reed-Sternberg (RS) cells that are set in a background of a mixed inflammatory infiltrate. Depending on the architecture of the infiltrate and the percentage of atypical cells, the histologic appearance is subclassified as type A, B, or C. The different histologic patterns do not seem to correlate with distinct clinical appearances. We report a case of lymphomatoid papulosis with unusual histologic features affecting a 14-year-old girl. The presence of anaplastic cells in a background of myxoid stroma closely resembled a sarcoma on histologic examination. PMID- 12544104 TI - Mucinous carcinoma of the skin with apocrine-type differentiation: immunohistochemical studies. AB - We investigated the pathway of differentiation in a case of mucinous carcinoma of the skin (MCS) arising on the right temple of a 70-year-old man. Histopathologic findings of the tumor showed evidence of apocrine-type differentiation such as distinctive decapitation secretion. Additionally, by employing a panel of antibodies, the immunohistochemical staining pattern of tumor cells was shown to be compatible with that of apocrine glands. Although the differentiation of this neoplasm remains controversial, the findings in our case suggest apocrine differentiation. PMID- 12544105 TI - The man behind the eponym: C. L. Pierre Masson. AB - This report traces the life of Pierre Masson and mentions some of his investigations. It summarizes Masson's seminal descriptions of glomus tumor and hemangioendotheliome vegetant intra-vasculaire (intravascular papillary endothelial hyperplasia), also known as Masson's pseudoangiosarcoma and the Masson lesion. Finally, we reprise Masson's view of the origins of melanocytic nevi and mention Masson's neuronevus. The literature on these subsequent to Masson's articles is not discussed. PMID- 12544106 TI - A basaloid neoplasm with ductal differentiation. PMID- 12544107 TI - Clinical pathological challenge: confluent and reticulate papillomatosis of Gougerot and Carteaud. PMID- 12544108 TI - The longer your telomeres, the larger your nevus? PMID- 12544109 TI - "Connective tissue nevus" and a serendipitous S-100 discovery. AB - Although desmoplastic melanoma is classically described as an indurated nodule on the head of an elderly patient associated with the histologic changes of an atypical intraepidermal melanocytic lesion accompanied by abnormal dermal spindled cells and rounded lymphoid infiltrates, it is capable of presenting in a variety of clinical and pathologic guises. Herein, a case with unusual clinical and histologic attributes is reported, which was initially misdiagnosed as a connective tissue nevus. PMID- 12544110 TI - Pictures of a unicorn? PMID- 12544111 TI - Renewed in a vision. PMID- 12544112 TI - Specialization in professional nursing. 1965. PMID- 12544114 TI - Upgrading systems design to reduce medication administration errors. PMID- 12544115 TI - Truthfulness. PMID- 12544116 TI - The role of nebulized opioids in managing terminal dyspnea: implications for the clinical nurse specialist. PMID- 12544117 TI - You don't have to leave your hospital system to be an entrepreneur. PMID- 12544118 TI - Redesign of clinical nurse specialist role course for distance education. Development, implementation, and outcomes. AB - The purpose of this article is to describe a collaborative effort to redesign a clinical nurse specialist curriculum for distance education among 4 regional state universities in southern Louisiana. The multisite approach was implemented to increase access to educational opportunities in advanced practice nursing for location-bound professional nurses. Course development, challenges to implementation, and success of the redesign are discussed. Additional topics described are online technical requirements and strategies for role socialization through distance learning. Success of the effort included an enhanced ability to recruit students to this specialty area. PMID- 12544119 TI - Acknowledging posttraumatic stress effects on health. A nursing intervention model. AB - Many people in our society have been exposed to overwhelming trauma, including abuse and assault. Posttraumatic stress can persist and become a chronic disorder, with behavioral and physiologic alterations affecting health across the lifespan. Often the etiologic role of trauma in a health problem remains undiscerned and unacknowledged. Acknowledging the effects of trauma is a caring intervention in itself, and it can lead to more effective healthcare and better relationships with patients. This article describes the process of acknowledging the effects of trauma in clinical reasoning, in dialogue with the patient, and in planning care and interventions. PMID- 12544120 TI - Clinical nurse specialist participation on a collaborative research project. Barriers and benefits. AB - Clinical nurse specialists are expected to contribute to the development of new nursing knowledge through research activities. Competing demands require constant juggling of time and priorities, and activities, such as research, often do not receive the attention they deserve. Research requires specific skills and knowledge and significant time expenditure. Collaborative research is an appropriate way for the clinical nurse specialist to both contribute to research and improve his or her research skills. This article discusses two pediatric critical care clinical nurse specialists' participation in a collaborative research team led by university faculty. The study was designed to reduce family uncertainty and stress and to increase staff nurse competency in providing family care. The ability of the clinical nurse specialists to contribute to the research team was enhanced by knowledge of the research process, good communication skills, and clinical expertise in the specific population of interest. Challenges for the clinical nurse specialists included limited experience in conducting clinical research and balancing clinical and research priorities. Both clinical nurse specialists and the university faculty benefited from this collaboration. The clinical nurse specialist, with expertise in a clinical specialty, is able to bridge the gap between the academic and clinical sites, making clinical research more likely. At the same time, working with expert researchers allows the clinical nurse specialist to increase his or her research skills. PMID- 12544121 TI - A description of clinical nurse specialist programs in the United States. AB - Changes have occurred in clinical nurse specialist educational requirements in recent years, and it is not known how programs have responded. The purpose of this descriptive survey was to identify the number of clinical nurse specialist programs in the United States, describe curricula, and examine enrollment and employment trends. This report contains data from 157 separate clinical nurse specialist programs/majors representing 139 different schools. The mean number of credit hours for semester- and quarter-based programs was 41.4 and 52.2, respectively. The mean number of clinical/practical clock hours was 416.2. Approximately half of the respondents (56%) used the National Association of Clinical Nurse Specialists to guide curricula. Nearly 66% of the programs had from 1 to 10 applicants during the previous year. The majority (68%) indicated that their graduates do not have difficulty finding employment after graduating. These findings indicate that efforts to revise curricula and clarify the CNS role must continue. PMID- 12544122 TI - NACNS responds to the National Council of State Boards of Nursing Uniform Advanced Practice Registered Nurse Licensure/Authority to Practice Requirements. PMID- 12544123 TI - Art, literature, and nursing phenomena. PMID- 12544124 TI - Comparison of I-123 and I-131 for whole-body imaging after stimulation by recombinant human thyrotropin: a preliminary report. AB - PURPOSE: Many studies have established I-123 whole-body scanning (WBS) as an alternative to I-131 WBS in the evaluation of differentiated thyroid cancer. Similarly, recombinant human thyroid-stimulating hormone (rhTSH) has been shown to be an effective alternative to thyroid hormone withdrawal in the stimulation of TSH levels for WBS. The purpose of the current investigation was to compare the ability of rhTSH-stimulated I-123 WBS to that of rhTSH-stimulated I-131 WBS in the evaluation of thyroid cancer. METHODS: The reports of 101 consecutive I 123 WBS and 101 consecutive I-131 WBS were analyzed. All patients had undergone near-total thyroidectomy, were considered to have a low risk for recurrence, and had rhTSH-stimulated serum thyroglobulin levels measured at the time of imaging. Findings of an examination were considered positive if either residual thyroid tissue was detected on imaging or the serum thyroglobulin level was greater than 2 ng/dl. RESULTS: The results of rhTSH-stimulated I-123 scans and thyroglobulin levels were concordant in 91 (90%) cases, whereas the results of rhTSH-stimulated I-131 scans and thyroglobulin levels were concordant in 85 (84%) cases. I-123 WBS detected 9 foci of disease in 6 patients, whereas I-131 WBS detected 10 foci of disease in 9 patients. CONCLUSION: The results of this preliminary investigation are promising regarding the utility of rhTSH-stimulated I-123 WBS as an alternate approach to rhTSH-stimulated I-131 WBS in patients at low risk for recurrence. PMID- 12544125 TI - Areolar-cutaneous "junction" injections to augment sentinel node count activity. AB - PURPOSE: The authors report on a modified lymphoscintigraphy protocol for increasing activity in the sentinel node (SN) through a specific technique (LymphoBoost). It consists of an areolar-cutaneous "junction" injection, using a very shallow, high-volume, high-specific-activity injection of 100% filtered Tc 99m sulfur colloid, as an adjunct to their standard protocol. MATERIALS AND METHODS: Results from a previously optimized protocol (group 1, n = 28) were compared with those from their new protocol (group 2, n = 85), which consisted of two sets of consecutively applied (within 12 to 20 minutes) injections: group 2A composed of perilesional and intradermal injections (similar to the previous group 1) followed by group 2B LymphoBoost injections within 12 to 20 minutes in the same patients. Regions of interest were drawn around the SN and the injection sites (IS) at the end of the studies to calculate the end-of-study SN:IS ratio for both group 1 and group 2 studies. The SN:IS ratio is generally independent of dose and is a measurement of the "efficiency" of getting activity from the IS to the SN. RESULTS: The mean SN:IS ratio in group 2 was 3.34 times greater than that in group 1 studies (P < 0.0005). The median SN:IS ratio was 3.53 times greater in the group 2 studies. Many cases showed a dramatic increase in SN counts before the LymphoBoost injection was even completed, with more than 5% of injected activity reaching nodes at the end of the study in some patients. Multiple different lymphatic pathways were noted, but all led to the same node(s). No significant disagreement between group 2A and group 2B results was noted. CONCLUSIONS: Areolar-cutaneous junction injections, performed under these conditions, augment SN activity dramatically in most patients. Hotter nodes provide several benefits, especially when next-day surgery is contemplated, and should also reduce the extent of dissection needed to remove the sentinel node. PMID- 12544126 TI - Calcium channel blockers: a potential cause of false-positive captopril renography. AB - PURPOSE: The purpose of the study was to review the causes of false-positive and false-negative captopril Tc-99m MAG3 renograms performed to evaluate renovascular hypertension at the authors' institution. METHODS: From May 2000 to January 2002, 77 patients were referred for evaluation of possible renovascular hypertension with captopril Tc-99m MAG3 renography. If the findings of the captopril study were abnormal, a baseline study was performed 3 days later. The captopril studies were retrospectively reviewed and correlated with available renal arteriography, magnetic resonance angiography, and duplex Doppler sonography studies and clinical follow-up. Renal artery stenosis greater than 70% on an arteriogram and 60% on Doppler studies or magnetic resonance angiography was considered positive for renovascular hypertension. RESULTS: There were five false-negative and four false-positive studies. The five patients who had the false-negative captopril studies underwent arteriography, which showed renal artery stenosis, and a stent was placed in two of them. The four false-positive captopril studies were proved by negative arteriograms in three and negative magnetic resonance angiography in one. Three patients with false-positive studies had bilateral renal function impairment after captopril and were taking calcium channel blockers at the time of the captopril study. One patient had a left nephrectomy and was also taking calcium channel blockers. CONCLUSION: Chronic intake of calcium channels blockers is a potential cause of bilateral symmetric false-positive captopril renography. PMID- 12544127 TI - Concomitant hyperparathyroidism and nonmedullary thyroid cancer, with a review of the literature. AB - PURPOSE: Primary hyperparathyroidism and concomitant thyroid cancer is a rare and complicated setting for diagnostic imaging. MATERIALS AND METHODS: The authors report the accidental finding of primary hyperparathyroidism in a patient with rapid enlargement of a thyroid nodule and the results of a literature review. RESULTS: Tl-201-Tc-99m subtraction scintigraphy correctly revealed the malignant nature of a large cold thyroid nodule and mediastinal parathyroid hyperplasia. In contrast, high-resolution ultrasound indicated a retrothyroidal hyperplastic parathyroid gland. Surgery followed the findings of the preoperative ultrasound and intraoperative biopsy, yet hyperparathyroid disease persisted. Repeated scintigraphy confirmed an ectopic parathyroid gland, which was resected from a paraesophageal location. Subsequently, hormone and calcium levels returned to normal and remained normal during a follow-up period of 3 years. A literature review revealed a prevalence of approximately 3% of nonmedullary thyroid cancer, which was found in patients operated on for primary hyperparathyroidism. Previous neck irradiation, especially in childhood, appears to be a risk factor for the development of both nonmedullary thyroid carcinoma and for primary hyperparathyroid disease. CONCLUSIONS: This case illustrates the need for clinical awareness of concomitant hyperparathyroidism and nonmedullary thyroid cancer and is substantiated with published case reviews. The preoperative scintigraphic localization of hyperfunctioning parathyroid tissue, although not advised as a routine procedure, may provide diagnostic information in addition to high-resolution ultrasound and intraoperative biopsy. In addition, scintigraphy can be useful even in the technically difficult setting of concomitant thyroid cancer. PMID- 12544128 TI - Scintigraphic findings in necrotizing myopathy. AB - Necrotizing myopathy is a rare syndrome associated with several causes, including non-small-cell lung carcinoma. The authors present a case of this infrequently occurring disease, describe an unusual scintigraphic pattern, and show tracer uptake localized in soft tissues rather than the skeleton. PMID- 12544129 TI - Radionuclide imaging in thoracic splenosis and a review of the literature. AB - Splenosis is defined as the autotransplantation of splenic tissue to abnormal locations after splenic injury. Although abdominal splenosis is the most common form and is usually discovered at laparotomy, thoracic splenosis is less common and occurs as an asymptomatic peripheral pulmonary nodule, incidentally discovered on a routine chest radiograph. Given the long interval between the initial trauma and its discovery, thoracic splenosis is rarely considered in the differential diagnosis of left-sided, pleural-based pulmonary nodules or masses. The diagnosis is often a result of surgery. The authors report an additional case of thoracic splenosis, review the literature, and discuss nonsurgical diagnostic methods. PMID- 12544130 TI - False-positive splenic imaging secondary to cavernous transformation of the portal venous system in non-Hodgkin's lymphoma. PMID- 12544131 TI - Rhabdomyolysis of bilateral teres major muscles. PMID- 12544132 TI - Scintigraphy of gluteal tendonopathy. PMID- 12544133 TI - FDG positron emission tomographic imaging of a large abdominal aortic aneurysm. PMID- 12544134 TI - Transposition of the great vessels manifested on radionuclide ventriculography. PMID- 12544135 TI - Early enhancing lesion seen on computed tomography consistent with focal nodular hyperplasia. PMID- 12544136 TI - Diagnosis of focal nodular hyperplasia with hepatobiliary scintigraphy using a modified SPECT technique. PMID- 12544137 TI - Massive intrathoracic toxic multinodular goiter treated with radioiodine. PMID- 12544138 TI - SPECT and planar imaging of a lingual thyroid gland. PMID- 12544139 TI - Elevated F-18 FDG uptake in the thymus in Graves' disease. PMID- 12544140 TI - The use of bone scintigraphy to evaluate metastatic calcification caused by end stage renal disease and secondary hyperparathyroidism. PMID- 12544141 TI - The heart or a tumor? Lung cancer in the lower left thorax can mimic myocardial FDG uptake on PET imaging. PMID- 12544142 TI - Renal ptosis and change of glomerular filtration values in the upright and supine positions. PMID- 12544143 TI - Extrarenal adult nephroblastoma. PMID- 12544144 TI - Unusual bone scan of a child with osteogenesis imperfecta. PMID- 12544145 TI - Ga-67 in Castleman's disease. PMID- 12544146 TI - Demonstration of necrotizing fasciitis on Ga-67 scans in a patient with systemic lupus erythematosus. PMID- 12544147 TI - Distinctive imaging evidence of brachial plexus neuritis demonstrated on Ga-67 citrate scintigraphy. PMID- 12544148 TI - Ring-like FDG uptake in acute cholecystitis. PMID- 12544150 TI - Decision support computer program for cancer pain management. AB - The purpose of the study was to develop an initial version of computer software that could assist nurses' decision making about cancer pain reported by women from diverse cultural groups. This cross-sectional study included two phases: (1) data collection and (2) development of computer software. Data were collected using an Internet survey and e-mail group discussions of 19 faculty members from 10 countries who were self-identified experts in oncology nursing. The data were analyzed using descriptive statistics and content analysis. The findings indicated ethnic, gender, geographic, and age differences in cancer pain descriptions. Based on the collected data, a decision support computer program for cancer pain management, including (1) a knowledge base generation module, (2) a decision-making module, and (3) a self-adaptation module, was developed. Based on the study findings, suggestions for future research and practice related to cancer pain and expert systems were proposed. PMID- 12544151 TI - Sepsis alert and diagnostic system: integrating clinical systems to enhance study coordinator efficiency. AB - Screening patients for clinical studies is time-consuming for researchers. Inefficiencies from human-based eligibility screening cause delay in scientific breakthroughs and are costly. We sought to determine the reliability of an automated computer-based real-time eligibility screening tool. A time-motion diary study was conducted in two university-based intensive care units using a cohort-controlled design. Time saved by automated eligibility screening and the positive and negative predictive values of the integrated eligibility screening system were compared with the gold standard of manual chart review. Sepsis Alert and Diagnostic System sensitivity and specificity were 82% and 95%, respectively. Positive and negative predictive values were 87.5% and 93%, respectively. During evaluation, Sepsis Alert and Diagnostic System saved a minimum of 137 minutes for the study coordinator. Sepsis Alert and Diagnostic System serves as a reliable tool for real-time eligibility screening in an intensive care unit setting. Time efficiencies through use of Sepsis Alert and Diagnostic System may translate into cost savings for funding agencies. The concept and methodology deployed in this study are applicable to any facility with electronic medical record capacity, as long as the data within that system are granular enough to support the specific query. PMID- 12544152 TI - Technology survey of nursing programs: implications for electronic end-of-life teaching tool development. AB - From an online survey of current technological capabilities of US undergraduate nursing programs, we found almost universal use of Microsoft Windows-based computers and Microsoft Office Suite software. Netscape and Microsoft Internet Explorer were the most popular browsers for Internet access. The survey also assessed faculty preferences for end-of-life care teaching materials and found that nurse educators preferred simple easy-to-use tools provided on CD-ROM or the Internet, with instructions provided via CD-ROM, the Internet, and demonstration workshops. Our findings have numerous implications for the development of electronic teaching materials for nursing. PMID- 12544153 TI - Modeling nursing care in health level 7 reference information model. AB - This article describes the attempt that was made to fit nursing knowledge into the Reference Information Model developed by Health Level 7 (HL 7 RIM). A part of nursing knowledge concerning pressure ulcer was used to show the possibilities of modeling nursing knowledge, terminology, and information in the Health Level 7 Reference Information Model. The purpose of this modeling work was to allow the integration of nursing information in electronic patient records and to ensure the interoperability and exchange of nursing information. Based on earlier reviews of pressure ulcer literature, an overview was made of relevant variables, coding, and information required for nursing care for pressure ulcer. Next, a use case was made and followed by modeling the required information in the Unified Modeling Language. It is possible to integrate nursing knowledge, terminology, and information requirements in the Health Level 7 Reference Information Model. PMID- 12544154 TI - Frequently asked questions about computer-adaptive testing. PMID- 12544155 TI - A practical way to create a library in a bibliography database manager: using electronic sources to make it easy. PMID- 12544156 TI - Jumping versus nonjumping anterior cruciate ligament injuries: a comparison of pathology. AB - OBJECTIVE: To compare pathology in knees with an anterior cruciate ligament (ACL) injury as a result of a jumping mechanism with knees injured from a nonjumping mechanism. DESIGN: This was a prospective study of 263 consecutive patients with a complete ACL tear and no subsequent giving way episodes nor reinjuries prior to undergoing an ACL reconstruction. The patients were placed into one of two groups: those with knees injured as a result of a jumping injury, and those with knees injured as a result of a nonjumping injury. SETTING: This study was performed at the Cleveland Clinic Foundation, Cleveland, Ohio. PATIENTS: Anterior cruciate ligament-deficient knees undergoing ACL reconstruction. Two hundred thirty-eight (91%) of the patients were injured while participating in sports. MAIN OUTCOME MEASURED: Intra-articular pathology documented prospectively at the time of arthroscopy. RESULTS: Knees injured as a result of a jumping mechanism had a higher incidence of medial (p = 0.05) and lateral (p = 0.03) meniscal tears and a lower incidence of medial collateral ligament injuries (p = 0.05). No difference in arthroscopic articular cartilage injuries was seen between the two groups. CONCLUSIONS: We believe that identifying jumping as a mechanism of ACL tears is important, since a jumping mechanism is associated with a significantly increased meniscus tear rate and may predispose this population to future degenerative changes. PMID- 12544157 TI - Degenerative changes in the ankle in former elite high jumpers. AB - OBJECTIVE: The object of this study was to find what degenerative changes are present in the ankle (talotibiofibular) joints in former elite high jumpers at least 10 years after their retirement from competition, whether there are differences between takeoff and swinging leg, and whether correlations between any particular training history parameters, former injuries, and the degenerative changes can be recognized. DESIGN: A cross-sectional case control study with matched controls for radiological outcomes SETTING: The Orthopedic Department at the University of Heidelberg, Germany. PARTICIPANTS: The subjects were 40 male high jumpers (required personal best at least 2.18m), featured in the lists of top athletes kept by the German Athletics Association (DLV) from 1972 to 1986. All radiological findings were compared with X-rays of male age, and BMI-matched controls. MAIN OUTCOME MEASUREMENTS: All underwent clinical and radiological examinations. In addition to eliciting data on the training history with the aid of a questionnaire, we assessed symptoms affecting the ankle by means of the Freiburg Ankle Score and the Kitaoka Score and scored the radiological findings according to Bargon and Scranton and MacDermott. RESULTS: Differences between takeoff and swinging leg were small (Freiburg takeoff leg 93/swinging leg 95, Kitaoka takeoff 89/swinging leg 93 points on a 100 points scale) in both clinical scores, but statistically significant (p < 0.005). The more jumps were performed during the active phase, the worse the radiological scores (r(s) = 0.4, p = 0.01). Radiological differences between takeoff and swinging leg were not found. Comparison with controls revealed no radiological differences between athletes and age- and BMI-matched men (all p-values >0.40). It was found that one takeoff and one swinging leg ankle was affected by grade 2 arthritis (Bargon). Scranton grade 3 was found in four takeoff and in three swinging legs. No further correlations with training history data were found. Athletes who reported injuries in the past tended to have poorer radiological scores, although statistically not significant. None of the athletes had an instable ankle. CONCLUSIONS: The talotibiofibular joints in former high jumpers showed only slight signs of wear and tear with no clinically relevant side-related differences; severe arthrosis with narrowing of the joint space was rare. The risk of arthrosis connected with high-jumping seems not to be elevated. PMID- 12544158 TI - Clinical diagnosis of Achilles tendinopathy with tendinosis. AB - OBJECTIVE: To evaluate sensitivity, specificity, reproducibility, and predictive value of palpation of the painful arc sign and of the Royal London Hospital test in 10 patients with Achilles tendinopathy and in 14 asymptomatic subjects. DESIGN: Test-retest study. SETTING: University teaching hospital. PARTICIPANTS: Ten male athletes on the waiting list for exploration of one of their Achilles tendons for tendinopathy of the main body of the tendon attended a special clinic. Each was invited to bring at least one athlete of the same sex in the same discipline aged within 2 years of themselves with no history and no symptoms of Achilles tendinopathy. A total of 14 controls were thus recruited. MAIN OUTCOME MEASURES: Pain and tenderness following performance of palpation, the painful arc sign, and the Royal London Hospital test. RESULTS: There were no statistically significant differences at the 5% level among the effects of investigator or between morning and afternoon measurements for any of the three measurement methods. There was no evidence of a difference of the three assessment methods (p > 0.05). When the three methods were combined, the overall sensitivity was 0.586 (confidence interval [CI], 0.469-0.741), and the overall specificity was 0.833 (CI, 0.758-0.889). CONCLUSIONS: In patients with tendinopathy of the Achilles tendon with a tender area of intratendinous swelling that moves with the tendon and whose tenderness significantly decreases or disappears when the tendon is put under tension, a clinical diagnosis of tendinopathy can be formulated, with a high positive predictive chance that the tendon will show ultrasonographic and histologic features of tendinopathy. PMID- 12544159 TI - The effects of home interferential therapy on post-operative pain, edema, and range of motion of the knee. AB - OBJECTIVE: We studied the effects of home interferential current therapy (IFC) on postoperative pain, range of motion, and edema in subjects undergoing anterior cruciate ligament (ACL) reconstruction, menisectomy, or knee chondroplasty. DESIGN: Randomized, double-blind, placebo-controlled prospective study. SETTING: A tertiary care outpatient orthopaedic clinic/ambulatory surgery center. SUBJECTS OR PARTICIPANTS: Eighty-seven subjects were separated into three groups based on their type of knee surgery and within each group randomized into a treatment or placebo group. INTERVENTIONS: All subjects received home IFC units. Subjects randomized to treatment group received a working IFC unit. Placebo subjects received units that were previously set to deliver no current. MAIN OUTCOME MEASUREMENTS: Post-operative edema at 24, 48, and 72 hours, and weeks 1-8; range of motion at 1, 3, 6, and 9 weeks; pain immediately after surgery, at 24, 48, and 72 hours, and weeks 1-7; and amount of pain medication taken at days 1-10 were compared between treatment and placebo groups. RESULTS: All IFC subjects reported significantly less pain and had significantly greater range of motion at all post operative time points. ACL and menisectomy IFC subjects experienced significantly less edema at all time points, while chondroplasty subjects experienced significantly less edema until 4 weeks postoperatively. CONCLUSIONS: These findings indicate that home IFC may help reduce pain, pain medication taken, and swelling while increasing range of motion in patients undergoing knee surgery. This could result in quicker return to activities of daily living and athletic activities. PMID- 12544160 TI - Assessment of cognitive recovery following sports related head trauma in boxers. AB - OBJECTIVE: To prospectively examine recovery of cognitive function within one month following subconcussive sports related head trauma. DESIGN: A prospective study of New York State licensed professional boxers who underwent testing of cognitive functioning before and after (within days, one week, and one month) a professional bout. SETTING: Male professional athletes recruited from the New York State Athletic Commission and local boxing gyms. PARTICIPANTS: Twenty-six licensed professional boxers were enrolled in the protocol. Data is presented on the 18 participants who completed testing on at least three of the four time points. INTERVENTIONS: Serial neuropsychological assessment before and after the athletes engaged in competition. MAIN OUTCOME MEASURES: Neuropsychological measures of cognitive functioning, including new learning and memory, information processing speed, and mental flexibility. RESULTS: A series of repeated measures MANOVAS revealed significant within subject differences across testing on complex information processing and verbal fluency. Post hoc analyses indicated significant differences between time 1 (baseline) and time 4 (one month post), with scores one month following the bout indicating significantly improved performance. Memory scores did not change significantly across testing; however, prior boxing exposure measured by total number of professional bouts was associated with poorer memory performance. CONCLUSIONS: Cognitive testing one month following participation in a professional boxing bout yielded scores suggestive of recovery to a level above the baseline. We conclude that baseline assessment taken during periods of intense training are likely confounded by other pre-bout conditions (i.e., sparring, rapid weight loss, pre-bout anxiety) and do not represent true baseline abilities. Instability of performance associated with mild head injury may complicate the interpretation of post-injury assessments. Practice effects may also confound the interpretation of serial assessments, leading to underestimation of the effects of sports related head trauma. Poorer cognitive performance was evident during the presumed recovery period in boxers with greater exposure to the sport (>12 professional bouts). This finding is consistent with reports of a cumulative effect of repetitive head trauma and the subsequent development of chronic traumatic brain injury. These data have implications for assessing recovery of function following head injury in players of other contact sports as well as determination of return-to-play following an injury. PMID- 12544161 TI - CogSport: reliability and correlation with conventional cognitive tests used in postconcussion medical evaluations. AB - OBJECTIVE: To determine the repeatability of a computerized cognitive test designed to monitor recovery from concussion and assist team physicians make return to play decisions (CogSport). To determine the correlation between CogSport and two conventional neuropsychological tests. DESIGN: Prospective, serial investigation of cognitive function. SETTING: Professional and semi professional Australian Football clubs and a university affiliated research institute in Melbourne, Australia. PARTICIPANTS: Three-hundred healthy young adults, including 240 elite athletes. MAIN OUTCOME MEASURES: Intra-class correlation (ICC) coefficients for CogSport performance measures administered serially. ICC between CogSport performance measures and conventional neuropsychological tests. Normative data for CogSport performance measures. RESULTS: Measures of psychomotor function, decision making, working memory and learning were highly reliable. Some measures also displayed high correlations with conventional neuropsychological tests of information processing and attention. Preliminary normative data is described. CONCLUSIONS: CogSport is a highly reliable cognitive function test when administered to healthy young adults and elite athletes. CogSport measures similar cognitive functions as conventional tests used commonly in concussion research. PMID- 12544162 TI - Medication use in athletes selected for doping control at the Sydney Olympics (2000). AB - During the Olympic Games held in Sydney in September, 2000 Doping Control was undertaken as specified in the International Olympic Code. During this process information about the medications taken by athletes was collected as a routine and formed part of the paperwork associated with a urine test. In their Post Games Report the World Anti-Doping Agency (WADA) recommended that the information about medications be collated with a view to assessing their use by athletes. Mandatory doping control for winners of events as well as random selection of athletes both during competition and out of competition allowed data to be collected about medications and supplements used by athletes. At the Doping Control Stations all competitors selected for a test, after providing a urine sample for analysis, were asked the same question: "what medications have you taken in the past three days?" The answer was to include all prescription drugs, over-the-counter medications, any other substances taken by mouth, injection, inhalation, ointment or by suppository, as well as vitamins, minerals, and all other supplements. This paper reviews the data from the 2758 Declaration Forms obtained at doping control. The prevalence of use of medications, the number used by an individual, and the pattern of use by these elite sports people were examined. The trends seen in this survey point to a dangerous overuse of nonsteroidal anti-inflammatory agents and an unnecessary overuse of vitamins in this population, while pointing out the increased prevalence of asthma and the dangers of drug interactions. OBJECTIVE: The main objective here is to review some of the medications used by athletes in the Olympic Games in Sydney 2000. DATA SOURCES: During these Games Doping Control was undertaken as specified by the International Olympic Committee. As well as a urine test, information about medications routinely taken was collected. Mandatory doping control for winners of events as well as random selection of athletes both during competition and out of competition required data to be collected about medications and supplements used by athletes as part of the sample collection protocol. At the Doping Control Stations all competitors selected for a test, after providing a urine sample for analysis, were asked the same question: "what medications have you taken in the past three days?" The answer was to include all prescription drugs, over-the counter medications, any other substances taken by mouth, injection, inhalation, ointment or by suppository, as well as vitamins, minerals, and all other supplements. DATA SELECTION: In this article we review the data from the laboratory copy of the 2758 Declaration Forms obtained at doping control. The cut down version of the Declaration Form submitted to the laboratory had all information identifying the athlete removed. Thus all information used in this article is completely anonymous. The prevalence of use of medications, the number used by an individual, and the pattern of use by these elite sports people were examined at the request of the IOC. CONCLUSIONS: In their Post-Games Report, the World Anti-Doping Agency (WADA) acting as independent observers of the anti doping process recommended to the IOC that the information obtained in the Athlete Declaration Forms concerning medications be collated with a view to assessing their use by athletes. The trends in their use seen in this survey point to an overuse of supplements as well as a dangerous overuse of drugs such as nonsteroidal anti-inflammatory agents together with multiple drug use emphasising the dangers of drug interactions and points out the increased prevalence of asthma in this population. PMID- 12544163 TI - The incidence, risk factors, and clinical manifestations of hyponatremia in marathon runners. AB - OBJECTIVE: To report on the incidence, identify the risk factors, and clarify the clinical manifestations of acute hyponatremia in marathon runners. DESIGN: An observational and retrospective case-controlled series. SETTING: The medical care area of the 2000 Houston Marathon. PATIENTS: Marathon finishers treated in medical area receiving intravenous fluids (N=55), including a more detailed analysis of 39 runners completing a retrospective questionnaire. MAIN OUTCOME MEASURES: Vital signs, serum electrolytes, and finish time were analyzed via ANOVA studies between all non-hyponatremic (NH: N=34)) and hyponatremic (H: N=21)) runners. Fluid intake, training variables, NSAID use, and Symptomatology were further analyzed to delineate all significant differences between groups. RESULTS: There were no significant differences in vital signs, training variables, or NSAID use between H and NH groups, although there was a trend towards the less experienced runners presenting with lower post-race sodium levels. H runners had lower potassium [K] (p=.04), chloride [Cl] (p<.001), and blood urea nitrogen [BUN] (p=.004) levels than NH runners. There was a significant inverse linear relationship between both finish time versus [Na] (r2 =.51) and total amount of fluid ingested versus [Na] (r2 =.39). The total cups of water (p=.004), electrolyte/carbohydrate solution (p=.005) and total amount of fluid ingested (p<.001) were significantly higher in H compared to NH runners and the degree of hyponatremia was related in a dose dependent manner. Vomiting was observed more frequently in H than NH runners (p=.03). CONCLUSION: 21 runners presented to the medical area of the Houston Marathon with hyponatremia (.31% of entrants). Excessive fluid consumption and longer finishing times were the primary risk factors for developing this condition. Vomiting was the only clinical sign differentiating hyponatremia from other conditions that induce exercise-associated collapse. PMID- 12544164 TI - Practical management: hamstring muscle injuries. PMID- 12544165 TI - Unusual pattern of injuries following knee hyperextension: a case report. PMID- 12544167 TI - The answer is on! A response to the initial lateral cervical spine film for the athlete with a suspected neck injury: helmet and shoulder pads on or off? PMID- 12544168 TI - Prehospital care of the spine-injured athlete: monograph summary. PMID- 12544169 TI - Psychiatric disorders in detainees. PMID- 12544170 TI - Psychiatric diagnosis in preschool children. PMID- 12544171 TI - Extraversion as a risk factor. PMID- 12544172 TI - Youth suicide and Hurricane Andrew. PMID- 12544173 TI - Treatment recommendations for the use of antipsychotics for aggressive youth (TRAAY). Part I: a review. AB - OBJECTIVES: To review the evidence for the safety and efficacy of nonpharmacological and pharmacological treatments for aggression in children and adolescents. METHOD: and searches (1990-present) were conducted for double-blind, placebo-controlled studies of atypical antipsychotics for aggression and for literature on the use of other pharmacological agents and psychosocial interventions for aggression. Case reports and adult literature regarding the safety of atypical antipsychotics were used where controlled data for youth were lacking. RESULTS: Controlled data on the treatment of aggression in youth is scarce. Psychosocial interventions may be effective alone or in combination with pharmacological treatments. Psychotropic agents (e.g., stimulants, mood stabilizers, beta-blockers) have also been shown to have limited efficacy in reducing aggression. Antipsychotics, particularly the atypical antipsychotics, show substantial efficacy in the treatment of aggression in selected pediatric populations. Atypical antipsychotics are generally associated with fewer extrapyramidal symptoms than are typical antipsychotics. CONCLUSIONS: Psychosocial interventions and atypical antipsychotics are promising treatments for aggression in youth. Double-blind studies should examine the safety and efficacy of atypical antipsychotics compared to each other and to medications from other classes, the efficacy of specific medications for different subtypes of aggression, combining various psychotropic medications, optimal dosages, and long-term safety. PMID- 12544174 TI - Treatment recommendations for the use of antipsychotics for aggressive youth (TRAAY). Part II. AB - OBJECTIVE: To develop treatment recommendations for the use of antipsychotic medications for children and adolescents with serious psychiatric disorders and externalizing behavior problems. METHOD: Using a combination of evidence- and consensus-based methodologies, recommendations were developed in six phases as informed by three primary sources of information: (1) current scientific evidence (published and unpublished), (2) the expressed needs for treatment-relevant information and guidance specified by clinicians in a series of focus groups, and (3) consensus of clinical and research experts derived from a formal survey and a consensus workshop. RESULTS: Fourteen treatment recommendations on the use of atypical antipsychotics for aggression in youth with comorbid psychiatric conditions were developed. Each recommendation corresponds to one of the phases of care (evaluation, treatment, stabilization, and maintenance) and includes a brief clinical rationale that draws upon the available scientific evidence and consensus expert opinion derived from survey data and a consensus workshop. CONCLUSION: Until additional research from controlled trials becomes available, these evidence- and consensus-based treatment recommendations may be a useful approach to guide the use of antipsychotics in youth with aggression. PMID- 12544175 TI - Early-onset schizophrenia in children with mental retardation: diagnostic reliability and stability of clinical features. AB - OBJECTIVES: To investigate the presentation of early-onset schizophrenia in children with mental retardation (MR) and the stability of clinical features over time. METHOD: A purpose-designed assessment protocol was developed based on best practice recommendations in the research literature. Diagnostic reliability, investigated in a group of 20 children with MR, was found to be very good (kappa = 0.87). Using this protocol, the clinical service identified 10 children with an initial diagnosis of schizophrenia. This group was then reassessed 2 years later. RESULTS: At the end of 2 years, 8 of the 10 children with schizophrenia still warranted this diagnosis. However, there was considerable instability in relation to certain diagnostic criteria, notably delusions and visual hallucinations. In all cases some improvements in severity were seen at follow-up. However, the eight whose schizophrenia was confirmed at follow-up showed major deterioration in cognitive level. CONCLUSIONS: It is possible to make reliable diagnoses of early-onset schizophrenia in people with MR, provided the assessment system is carefully structured and pays attention to all the important sources of information. However, symptoms should be reviewed frequently, particularly those that cannot be substantiated by patient interview. PMID- 12544176 TI - A prospective, open-label trial of olanzapine in adolescents with schizophrenia. AB - BACKGROUND: Olanzapine is an atypical antipsychotic that has efficacy in adults with psychotic disorders. This preliminary study examined the effectiveness of olanzapine in adolescents with schizophrenia or its related conditions. METHOD: Adolescents aged 12-17 years (inclusive) with a diagnosis of schizophrenia, schizoaffective, or schizophreniform disorder were enrolled in this 8-week, open label, outpatient study. The Positive and Negative Syndrome Scale (PANSS), the Clinical Global Impressions Scale (CGI), and the Children's Global Assessment Scale (CGAS) were administered as outcome measures. Extrapyramidal side effects were assessed at each visit. Olanzapine was initiated at a dose of 2.5 mg/day and could be increased to a maximum total daily dose of 20 mg. RESULTS: Sixteen participants with a mean age of 13.8 (SD = 1.5) years were treated. Significant improvements were found in the PANSS, CGI severity, and CGAS scores. Reductions in both positive and negative symptoms were found. Increased appetite and sedation were the most frequently reported side effects. Two subjects required treatment for extrapyramidal side effects. CONCLUSIONS: Psychotic symptoms significantly improved during study. Overall, olanzapine was well tolerated. Future studies are needed to confirm these findings, to assess long-term treatment outcomes, and to compare the effectiveness of olanzapine with that of other antipsychotics. PMID- 12544177 TI - Adult outcome of social function in adolescent-onset schizophrenia and affective psychosis. AB - OBJECTIVE: To examine and compare the adult outcome in a representative sample of hospitalized adolescent-onset psychoses including occupational and social aspects. METHOD: A total of 81 patients with a first episode of early-onset psychosis (before age 19 years) presenting to the University Hospital of Lund, Sweden, between 1982 and 1993 were followed up an average of 10.5 years (range 5.1-18.2) after admission. Initial diagnosis was assessed from records and consisted of DSM-IV schizophrenia (n = 32), schizoaffective disorder (n = 7), bipolar disorder (n = 25), and major depressive disorder with psychotic features (n = 17). All could be traced and assigned a major outcome group. RESULTS: Early onset schizophrenia spectrum disorder suffered a chronic course with a poor outcome in 79% of the cases, while early-onset affective psychosis in 74% showed a good or intermediate outcome. The poor outcome (26%) in the affective group was connected to mental retardation in 7% and to progression to a schizoaffective disorder in 12%. A particularly severe outcome was seen for schizophrenia spectrum patients with a family history of nonaffective psychosis. CONCLUSIONS: Early-onset schizophrenia spectrum disorder showed a severe course while affective psychoses had a much more benign functional outcome. PMID- 12544178 TI - Child temperament, ADHD, and caregiver strain: exploring relationships in an epidemiological sample. AB - OBJECTIVE: To investigate the relationship between child-reported dimensions of temperament and attention-deficit/ hyperactivity disorder (ADHD), as well as other indicators of child psychopathology, including disruptive disorders, depression, and anxiety. It also examined whether difficult child temperament scores independently predicted caregiver strain. METHOD: A school-district-wide, two-phase screening design (response rate 70% for phase 2) identified elementary school children at high risk for ADHD. Two hundred high-risk children and their parents completed standardized instruments to assess child temperament, diagnoses of disruptive disorders, children's symptoms of anxiety and depression, and caregiver strain. Relationships were examined using analysis of variance, correlations, and multivariate prediction models, adjusting for child sociodemographic characteristics and psychopathology. All estimates were weighted for sampling design and differential participation. RESULTS: Combined subtype ADHD was associated with lower scores on task orientation and higher scores on general activity level. Depressive symptoms correlated significantly with all but one difficult temperament dimension, in a pattern consistent with clinical symptoms of depression. Child temperament did not vary by ADHD treatment status. Among these high-risk children, maternal caregiver strain experiences were increased by male gender, inattention symptoms, and oppositional defiant disorder, but not by difficult temperament scores. CONCLUSION: This study provides support for Graham and Stevenson's hypothesis of continuity between specific temperament traits and certain child psychiatric disorders, namely ADHD and depressive disorders. PMID- 12544179 TI - Correlates of comorbid psychopathology in children with ADHD. AB - OBJECTIVE: To investigate correlates of internalizing and externalizing psychopathology in a clinical sample of children with attention deficit/hyperactivity disorder (ADHD). METHOD: Parent and teacher Child Behavior Checklists were administered to 300 children with ADHD to ascertain comorbid symptoms. Based on previous research, a seven-step hierarchical regression analysis was developed. Six hierarchical regression analyses were conducted with either parent or teacher Child Behavior Checklist aggression, delinquency, or anxious/depressive comorbid symptoms as dependent measures. RESULTS: Controlling for (1) variables known to increase risk for ADHD child psychopathology and (2) estimated duration of ADHD, our results suggest that the presence of comorbid symptoms is influenced by age of onset. An early age of onset of ADHD was correlated with a greater rate of parent-reported child aggressive symptoms, and a later age of onset was correlated with a greater rate of parent-reported child anxious/depressive symptoms. Elevated levels of comorbid externalizing and internalizing symptoms are associated with greater ADHD symptom severity. CONCLUSIONS: Comorbid externalizing and internalizing symptoms are correlated with age of ADHD onset and are related to the severity of clinical presentation in a referred sample of children with ADHD. PMID- 12544180 TI - Conduct problems and symptoms of sleep disorders in children. AB - OBJECTIVE: Conduct problems and hyperactivity are frequent among children referred for sleep-disordered breathing (SDB), restless legs syndrome, or periodic leg movements during sleep (PLMS), but children not referred to sleep centers have received little study. METHOD: Parents of children aged 2 to 14 years were surveyed at two general clinics between 1998 and 2000. A Pediatric Sleep Questionnaire generated validated scores for SDB and PLMS. The Conners Parent Rating Scale (CPRS-48) produced an age- and sex-adjusted Conduct Problem Index (CPI) and Hyperactivity Index. RESULTS: Parents of about 1,400 children were approached; those of 872 (62%) completed the surveys. Bullying and other specific aggressive behaviors were generally two to three times more frequent among 114 children at high risk for SDB than among the remaining children. An association between high CPI and SDB scores (p <.0001) retained significance after adjustment for sleepiness, high Hyperactivity Index, stimulant use, or PLMS scores. Analogous results were obtained for the association between high CPI and PLMS scores. CONCLUSIONS: Conduct problems were associated with symptoms of SDB, restless legs syndrome, and PLMS. Although these results cannot prove a cause-and effect relationship, assessment for sleep disorders may provide a new treatment opportunity for some aggressive children. PMID- 12544181 TI - Treatment effects of methylphenidate on behavioral adjustment in children with mental retardation and ADHD. AB - OBJECTIVE: The effects of stimulant medication treatment were investigated in children with mental retardation (MR) and attention-deficit/hyperactivity disorder (ADHD). METHOD: Parent and teacher behavioral ratings and reports of side effects were obtained for children (N = 24, mean age = 10.9 years, SD = 2.4) during a placebo-controlled, double-blind, crossover treatment trial with 0.15 mg/kg, 0.30 mg/kg, and 0.60 mg/kg b.i.d. dosages of methylphenidate. RESULTS: The most significant improvements occurred at the 0.60 mg/kg methylphenidate dose for teacher ratings of inattention (p =.024), hyperactivity (p <.001), aggression (p <.001), and asocial behavior (p =.009). No significant improvements, relative to placebo, occurred at the 0.15 mg/kg dosage. Of interest, nearly all significant medication-related behavioral improvements were detected by teachers. However, parents were sensitive raters of side effects, noting more sleeping problems and loss of appetite at the 0.60 mg/kg dose compared with placebo. CONCLUSIONS: These results suggest that symptoms of ADHD can be treated successfully in children with ADHD/MR, and consistent with MTA study results, higher doses were most effective. Furthermore, these improvements were not accompanied by increases in symptoms such as staring, social withdrawal, or anxiety. PMID- 12544182 TI - Onset of major depressive disorder among adolescents. AB - OBJECTIVES: To examine the association between parental affective disorders and psychoactive substance use disorders and the onset of major depressive disorder (MDD) among adolescents and young adults and to determine whether this association is affected by stressful life events, family cohesion, self-esteem, or gender. METHOD: Prospective cohort study of 804 adolescents, aged 11-17 years, and their parents who were followed for seven consecutive years. The sample was drawn from the Minneapolis-St. Paul metropolitan area. Parental diagnoses were based on Structured Clinical Interview for administered during study screening stage. Diagnoses of MDD and age of onset were based on Composite International Diagnostic Interview 2.1 administered during final year of data collection. RESULTS: Of the correlates examined, only parental affective disorders, low self esteem, and gender were significantly related to the onset of MDD. Females were twice as likely as males to experience MDD. CONCLUSIONS: The direct association between parental affective disorders and MDD onset was not affected by family cohesion, self-esteem, or stressful life events; thus more research is needed on other factors that may affect this association, such as genetic factors or other family- and intrapersonal-based variables. PMID- 12544183 TI - Major depression and conduct disorder in a twin sample: gender, functioning, and risk for future psychopathology. AB - OBJECTIVE: Major depression (MDD) and conduct disorder (CD) co-occur in adolescents at rates higher than would be expected by chance. This study described the functioning of adolescents with histories of these disorders and examined whether these patterns of association differed by gender. METHOD: Subjects with a lifetime diagnosis of MDD and/or CD were selected from a sample of 17-year-old twins; control subjects had no history of either disorder. The domains of school success, substance dependence, peer relationships, and age of first sexual intercourse were examined. RESULTS: Overall, each disorder separately and especially both disorders together related to increased maladjustment in the domains of school success and substance dependence. For school behavior problems, nicotine dependence, and drug dependence, the combination of MDD and CD related to particularly problematic functioning. Results were similar for males and females. Longitudinal data indicated that the occurrence of these disorders by late adolescence was predictive of subsequent depression and antisocial behavior in early adulthood. CONCLUSIONS: The combination of CD and MDD relates to more serious maladjustment, especially relating to school success and substance dependence, than would be expected given the adjustment associated with each disorder alone. PMID- 12544184 TI - Correlates of suicide risk in juvenile detainees and adolescent inpatients. AB - OBJECTIVE: To compare correlates of risk for suicidal behavior in juvenile detainees with those in another high-risk group, adolescent psychiatric inpatients. METHOD: Eighty-one adolescents in a short-term juvenile detention center were contrasted with a matched group of 81 adolescent psychiatric inpatients on a clinical assessment battery of established instruments including a measure for risk of suicidal behavior. RESULTS: Juvenile detainees and adolescent psychiatric inpatients reported similar levels of distress on measures of suicide risk, depression, impulsivity, and drug abuse. After controlling for depression, impulsivity and drug abuse remained significantly associated with suicide risk scores in the juvenile detention group, but did not in the psychiatric contrast group. For depressed female inpatients, hopelessness added significantly to the prediction of suicide risk scores. CONCLUSIONS: Correlates of risk for suicidal behavior in juvenile detainees may differ from those in other high-risk groups. Results suggest that it may be helpful to examine impulsivity and history of drug abuse when assessing suicide risk for detained adolescents. Further study of juvenile detainees as a separate high-risk group is warranted to better determine the nature and extent of risk. PMID- 12544185 TI - Spanish-language services assessment for children and adolescents (SACA): reliability of parent and adolescent reports. AB - OBJECTIVE: To assess test-retest reliability of the service utilization screening section of the Services Assessment for Children and Adolescents (SACA) interview among Spanish-speaking parents and adolescents, correspondence between parent and adolescent reports, and the correlation between reliability and participants' demographic and service use characteristics. METHOD: The English SACA was translated and administered from September 1999 through January 2000 in Los Angeles County, California, on two separate occasions to eligible parents with a child (4-17 years old) who was a client of a local public mental health authority. Adolescents of these parents (12-17 years old) were also interviewed. Reliability was measured by the kappa statistic. RESULTS: Adult and adolescent reports about lifetime and previous year service setting use exhibited good reliability, but concordance of parents and adolescents did not. Children's service utilization appears to be correlated with reliability of parent reports, and child gender appears to be correlated with reliability of adolescent reports. CONCLUSION: The SACA appears to be a useful tool for screening Spanish-speaking families about child and adolescent mental health service use. These findings must be considered preliminary until replicated in a larger sample of culturally diverse Spanish-speaking families. PMID- 12544186 TI - Between-groups factorial designs: analysis and interpretation. PMID- 12544187 TI - Genetics of childhood disorders: XLVII. Autism, part 6: duplication and inherited susceptibility of chromosome 15q11-q13 genes in autism. PMID- 12544188 TI - What do eosinophils tell us in biopsies of patients with inflammatory bowel disease? PMID- 12544189 TI - Management of pain in chronic pancreatitis: medical or surgical. PMID- 12544190 TI - Hepatitis C and alcohol. PMID- 12544191 TI - Endoscopic ultrasound and upper gastrointestinal disorders. AB - Endoscopic ultrasound (EUS) plays a vital role in management of upper gastrointestinal disorders, particularly cancer of the esophagus, pancreas, stomach, lung (via transesophageal mediastinal staging), and bile duct. Endoscopic ultrasound has also been valuable in detection of early chronic pancreatitis (CP). In cancer of the esophagus, the primary role of EUS is to determine whether disease is localized (T1-2, N0) and appropriate for surgery, locally advanced (T3-4, N1, M1a) (which may benefit from chemoradiation with or without surgery), or metastatic. Pancreatic and bile duct cancers are more complex given the controversy over portal vein resection. In centers that resect tumors invading the portal venous system, the role of EUS is limited to tissue confirmation or identification of metastases to the liver or distant lymph nodes. In centers that do not resect the portal vein invasion, EUS plays an important role in local staging. In lung cancer, EUS is emerging as an accurate, nonsurgical alternative to staging the mediastinum through EUS fine-needle aspiration. Endoscopic ultrasound has an important role in diagnosing CP because of its high degree of sensitivity. This has also led to controversy over whether EUS can overdiagnose CP. For these reasons, we recommend the use of a high threshold for EUS and that CP be diagnosed in conjunction with other standard tests (endoscopic retrograde cholangiopancreatography, pancreatic function tests). PMID- 12544192 TI - Use of probiotics in the treatment of inflammatory bowel disease. AB - The gut flora plays a fundamental role in maintaining normal intestinal function. A disturbance of this flora, or the host response to this flora, has been clearly demonstrated to play a role in the pathogenesis of inflammatory bowel disease (IBD). This has led to attempts to modify the bacterial flora with "live non pathogenic organisms that confer health benefits by improving the microbial balance," otherwise known as probiotics. Recent attention has focused on this potential strategy to treat or prevent IBD. The potential therapeutic benefit is enhanced by the natural and apparently safe approach that probiotics offer. Animal models of colitis have provided the proof of principle that probiotics can prevent and treat established intestinal inflammation. Controlled clinical studies have demonstrated the efficacy of probiotics in the maintenance of remission of pouchitis, prophylaxis of pouchitis after the formation of an ileoanal reservoir, maintenance of remission of ulcerative colitis, and treatment of Crohn's disease. However, large controlled trials are needed to definitively establish the place for probiotics in the treatment of IBD and resolve issues such as the dose, duration, frequency of treatment, and use of single or multiple strains. Research is focusing on establishing the mechanism of action, so that treatments with individually tailored properties are developed and innovative approaches are explored. PMID- 12544193 TI - Immunohistochemical study of intestinal eosinophils in inflammatory bowel disease. AB - BACKGROUND: Eosinophil accumulation and activation are characteristic features of inflammation in allergic diseases and in host defense against parasites. GOALS: To investigate the involvement of eosinophils in inflamed and noninflamed mucosa of patients with inflammatory bowel disease (IBD). STUDY: Specimens of inflamed colonic mucosa from 15 patients with ulcerative colitis (UC) and inflamed and noninflamed colonic mucosa from 15 patients with Crohn's disease (CD) were submitted to histologic and immunohistochemical studies. Twelve patients with irritable bowel syndrome were studied as controls. Sirius red was used to label eosinophils in tissue. EG1, EG2, and anti-hIL-5 were used as primary antibodies in an indirect alkaline phosphatase-labeled immunostaining protocol. Both positive and negative lamina propria cells were assessed by a quantitative grading system and the results expressed as cell numbers per mm. RESULTS: Increased proportions of eosinophils stained with Sirius red, EG1, EG2, and anti hIL-5+ cells were found in the colon of patients with UC and in inflamed and noninflamed colon of CD patients as compared with controls. Crohn's disease patients showed increased proportions of EG1+ and EG2+ cells as compared with those with UC. Increased proportions of IL-5+ cells were detected in UC patients as compared with those with CD. CONCLUSION: Quantitative eosinophil alterations and IL-5+ cells may indicate enhanced cellular activation with degranulation, which is implicated in the pathogenesis of IBD. Increase in IL-5+ cells may reflect a predominant local Th2 response in UC as compared with CD. PMID- 12544194 TI - Impact of imprint cytology in detecting short segment Barrett's esophagus. AB - BACKGROUND: Short-segment Barrett's esophagus (SSBE) is defined by the presence of columnar-appearing mucosa in distal esophagus (involving less than 2 to 3 cm), with intestinal metaplasia on biopsy. Its potential to develop dysplasia and cancer may require a surveillance program with better diagnostic tools to detect intestinal metaplasia. GOALS: To investigate the role of imprint cytology as a diagnostic tool either alone or combined with histology in SSBE. STUDY: Seventy nine patients (46 men, 33 women) with SSBE diagnosed during elective upper gastroscopy were included. Patients with serrated z-line with short tongues of pink mucosa and patients with a circular non-serrated z-line that extended less than 2 cm above the esophagogastric junction were biopsied on four quadrants just distal to z-line. Four slides of imprint preparation (including 1, 2, 3, and 4 touching of each biopsy specimen) was made for cytologic examination. Hematoxylin and eosin and Alcian blue staining for histologic examinations and Alcian Blue for cytologic evaluations were used to find evidence of intestinal metaplasia. RESULTS: Intestinal metaplasia was detected in 15 (19%), 21 (27%), and 30 (38%) patients by histologic examination with hematoxylin and eosin alone, by Alcian blue alone, and by histologic plus cytologic examination with Alcian blue, respectively. Nine patients with negative histologic but positive cytologic results were positive for intestinal metaplasia when they were reevaluated after further sectioning and staining. Sensitivity of imprint cytology alone was 53%. When imprint cytology was combined with the histologic evaluation, the prevalence of intestinal metaplasia increased from 27% to 38% (p < 0.05). CONCLUSION: Imprint cytology might be a complementary diagnostic tool for histology in detecting patients with SSBE. PMID- 12544195 TI - An association between Helicobacter pylori infection and serum vitamin B12 levels in healthy adults. AB - GOALS: To determine whether serum vitamin B12 levels in non-vitamin B12 deficient healthy adults correlate with serological evidence of H. pylori infection. BACKGROUND: An association between H. pylori infection and vitamin B12 deficiency has been recently reported. STUDY: 133 adults, presenting to a community based primary care clinic who met the following exclusion criteria; history of H. pylori eradication or antacid use, liver disease, inflammatory bowel disease, previous gastrointestinal surgery, a vegetarian diet or multivitamin supplementation were studied. Blood was drawn for a complete blood count, serum vitamin B12, gastrin, folic acid and H. pylori IgG antibodies. Subjects with vitamin B12 < or = 145 ng/mL (deficient range) were excluded. RESULTS: Of 133 subjects 96 (72.2%) were seropositive for H. pylori IgG antibodies (HP+). Age of HP(+) subjects did not differ from that of seronegative subjects (HP-); 52.8 +/- 1.6 mean +/- SE versus 49.2 +/- 2.9 ( = NS). Prevalence of HP seropositivity was significantly higher among subjects with borderline (>145-180 pg/mL) or low normal (>180-250 pg/mL) vitamin B12 levels than among those with vitamin B12 > 250 pg/mL; among 25 subjects with vitamin B12 > 145-180 pg/mL 92% were seropositive and among 47 subjects with vitamin B12 > 180-250 pg/mL 89% were seropositive as compared with 31/61 (51%) of subjects with B12 > 250 pg/mL, Fisher exact test < 0.0001. Vitamin B12 levels did not correlate with age (r = 0.07). Gastrin levels (pg/mL) did not differ significantly between groups; 70.2 +/- 5.8 in HP(+) versus 56.0 +/- 12.4 in HP(-). CONCLUSIONS: The higher prevalence of H. pylori infection among subjects with serum vitamin B12 levels that are within the lower end of the normal range suggests a causal relationship between H pylori infection and vitamin B12 levels in healthy adults. PMID- 12544196 TI - Seroprevalence of Helicobacter pylori antibodies in medical students and residents in Recife, Brazil. AB - BACKGROUND: The epidemiology of Helicobacter pylori (Hp) still constitutes a relevant subject of investigation. GOAL: The goal of this study was to determine the seroprevalence rate of Hp antibodies in medical students and residents, and its relationship with medical practice, age, gender and parents' formal education level. STUDY: The study was carried out from March 1999 to March 2000, on a population of 1,119 medical students and residents, from which a randomized and stratified sample was drawn. Each stratum comprised students attending the basic training level, students attending the professional training level, and residents. Independent samples were calculated for each group, making a total of 405 subjects. Each participant was interviewed using a questionnaire. After the interview, serum samples were collected and stored at -20 degrees C for detecting IgG antibodies against Hp by an enzyme-linked immunosorbent assay. RESULTS: A 31.4% seroprevalence rate of Hp antibodies was observed, which significantly increased during the graduation course according to the medical practices, ranging from 23.4% in students up to 38.6% in residents. Seroprevalence rate increased according to the age (p = 0.002), where older individuals had spent more time in practical activities. Seroprevalence was higher in males and was smaller among those whose parents had a university degree. CONCLUSION: The seroprevalence rate of Hp antibodies amongst medical students and residents increases from the basic up to residence level, suggesting that contact with patients during clinical practice may constitute a risk factor for acquiring Hp infection. PMID- 12544197 TI - Survey of the opinions, knowledge and practices of surgeons and internists regarding Helicobacter pylori test-and-treat policy. AB - BACKGROUND: Helicobacter pylori infection is highly prevalent and may cause disease in 10% to 15% of infected individuals: duodenal and gastric ulcer, gastric cancer and gastric lymphoma. Guidelines for a test-and-treat policy have been published by the European H. pylori working group in Maastricht. GOALS: To assess the current approach to H. pylori-related diseases, indications for testing and eradication therapy, among internists and surgeons of a University affiliated medical center. STUDY: A 15-items questionnaire was formulated according to the Maastricht II test-and-treat recommendation and distributed among the physicians of 6 internal medicine departments and the department of surgery. The questionnaires were completed anonymously at the department's staff meeting, under the supervision of the head of the department. All questions required a yes/no answer; the maximum possible score was 15. The relative ratio (RR) of correct answers was calculated for every question and by subgroups as follows: all participants, internists, surgeons, experts and residents in internal medicine and surgery. RESULTS: The response rate was 68% (55 out of 80 physicians). Total score for all participants was 10.9 (RR = 0.73), significantly lower than the maximum score of 15 (RR = 1.00). The internists had significantly higher average score than the surgeons for 2 questions: causative agent in gastric cancer (p = 0.003) or gastric lymphoma (p = 0.003), 1 question about triple therapy, PPI, penicillin and azythromycin being not recommended (p = 0.022), and for 1 question of test-and-treat policy: should this strategy apply to family members of gastric cancer patients (p = 0.045). Only 56% of the internists and 21% of surgeons knew what MALToma is, and the correlation with H. pylori infection. CONCLUSION: Knowledge of Surgeons and Internists regarding infection and correlation with diseases or test and treat policy should be improved. PMID- 12544198 TI - Collagenous sprue associated with an extensive T-cell lymphoma. AB - A 79-year-old woman developed collagenous sprue, a rare small intestinal mucosal disorder. Later, extensive T-cell lymphoma was documented, a neoplasm known to complicate celiac disease. Although the precise relationship of collagenous sprue to celiac disease has been debated and remains controversial, the findings here provide additional evidence that collagenous sprue and celiac disease are closely linked. In the past, long-term survival with collagenous sprue may have been compromised due to severe pan-malabsorption. With improved treatment measures, including modern nutritional support, it is likely that there will be an increased opportunity in future for improved appreciation of the complications of collagenous sprue, specifically, lymphoma. PMID- 12544199 TI - Perianal fistulae caused by Mycobacterium fortuitum. AB - Mycobacterium fortuitum is a rapidly growing Mycobacterium , which usually colonizes the soil, dust and water. It commonly causes skin and soft tissue infections especially in patients who have preceding trauma. We report a case of perianal fistulae caused by M. fortuitum. PMID- 12544200 TI - Liver disease in hereditary hemorrhagic telangiectasia. AB - Hereditary hemorrhagic telangiectasia (HHT), also known as Osler-Weber-Rendu disease, is an hereditary disorder that results in fibrovascular dysplasia with the development of telangiectasias and arteriovenous malformations. It predominantly involves the skin, mucous membranes, viscera, lungs, and brain. Hereditary hemorrhagic telangiectasia shows great genetic heterogeneity, and its phenotypes have been classified based on the recently identified mutated genes: endoglin (HHT-1) and activin-like kinase receptor-1 (HHT-2). Other families with phenotypic HHT do not bear these mutations; therefore, other genes are probably involved as well. Liver involvement is reported in up to 30% of persons affected by HHT. Large arteriovenous malformations in the liver can lead to significant complications, including high-output congestive heart failure, portal hypertension, hepatic encephalopathy, biliary ischemia, and liver failure. Embolization of large arteriovenous malformations in the liver remains controversial; however, liver transplantation can successfully eradicate these complications. PMID- 12544201 TI - Chronic pancreatitis. Long-term pain relief with or without surgery, cancer risk, and mortality. AB - To determine the natural history of chronic pancreatitis (CP), we retrospectively studied 193 consecutive patients who had at least one hospitalization for the control of pain or a complication of CP by examining the hospital records and by using a standard questionnaire. Alcohol (66%) was the major cause of CP and the cause was unknown in 21%. Pain was the presenting symptom in 93%. Pancreatic calcification was observed in 41% (alcoholic 54% vs. nonalcoholic 19%; OR = 6.7, CI = 2.7, 14.3; p < 0.0001). Diabetes (28%), malabsorption (16%), pseudocysts (21%) and pancreatic (3%) or extrapancreatic malignancy (5%) were the main complications. 43% had surgical intervention for pain relief, 10% had either endoscopic sphincterotomy or surgical sphincteroplasty and 16% had surgery for complications. Surgical or endoscopic intervention was more commonly performed in nonalcoholics compared with alcoholics (OR = 12.8, CI = 3.6, 53.9; p < 0.0001). However, if sphincterotomy and sphincteroplasty were excluded, the total number of surgical procedures for pain relief was similar in both groups. Complete follow-up information was available in 107 patients with a mean duration of follow-up of 10 years (range, 1-28 years); 27 patients died during the follow-up; 5, 10 and 15 year mortality was 14%, 18% and 20% respectively. The mortality was significantly higher in patients with alcoholic CP than in nonalcoholic CP (35% vs. 10%; OR = 1.4, 18.7; p = 0.005). Of the 80 patients who were alive and had complete long-term follow-up, pain improved in 62 patients, remained unchanged in 17 and worsened in one. Pain improved in 34 of 41 (83%) patients who had surgical intervention for pain, 7 of 9 patients (78%) who had surgery for complications, 4 of 7 (57%) who had sphincter ablation and 17 of 23 patients (74%) who had nonprocedural treatment. Long-term pain relief was similar in patients with alcoholic and nonalcoholic pancreatitis. PMID- 12544202 TI - Clinical features of acute acalculous cholecystitis. AB - BACKGROUND: Acute acalculous cholecystitis (AAC) tends to have a fulminant course and be associated with critically ill diseases, there have been reports of AAC without any risk factors but good prognosis. GOALS: To assess the risk factors, clinical features and prognosis of AAC. STUDY: All patients who had a cholecystectomy due to acute cholecystitis at Pundang Jesaeng General Hospital during a 43-month period were prospectively enrolled. AAC was defined by ultrasonographic, intraoperative and pathologic findings of acute cholecystitis without evidence of gallstones. Clinical features and pathologic findings were analyzed and outcome was assessed. RESULTS: 156 patients with acute cholecystitis were enrolled and 14% (22 of 156) met the criteria of AAC. Fifteen (68%) of the patients with AAC were male and the average age was 63 year old. Twenty patients were presented with AAC as outpatients of whom seven of them (35%) had atherosclerotic vascular disease. Laparoscopic cholecystectomy was performed in 126 patients (80.8%) with acute cholecystitis but was possible in only 12 patients (54.5%) with AAC. AAC was associated with a high incidence of gangrene (59%) but no patients died of acute cholecystitis. CONCLUSIONS: We conclude that AAC frequently occurs in elderly male outpatients without critical illness and gangrene is common but the prognosis is better than reported previously. PMID- 12544203 TI - Effect of hepatitis C virus infection and abstinence from alcohol on survival in patients with alcoholic cirrhosis. AB - GOALS: We assessed the effect of HCV infection and abstinence from alcohol on survival in a cohort of patients with alcoholic cirrhosis. BACKGROUND: Hepatitis C virus (HCV) infection may be an important cofactor for liver disease in chronic alcoholics. STUDY: The study population consisted of 213 patients with the diagnosis of alcoholic cirrhosis, 72 of these patients were infected by HCV. Complete alcohol abstinence after diagnosis of alcoholic cirrhosis was recorded in 86 patients. The reference group consisted of 89 patients with anti-HCV positivity who had never consumed alcohol. Survival was analyzed by the Kaplan and Meier method and predictors of survival by the Cox's multiple regression model. RESULTS: HCV infection was not a determinant factor for survival in alcoholic cirrhosis. Age and Child-Pugh grade at the time of diagnosis of cirrhosis and persistence of alcohol consumption after diagnosis were independent predictors of poor outcome. The cumulative survival curve in abstinent alcoholics was significantly different from that of alcoholics who maintained the same pattern of alcohol consumption (log-rank = 4.30, p = 0.0381). Moreover, the cumulative survival in patients with anti-HCV-positive cirrhosis who stopped drinking after diagnosis was similar to that in patients with HCV-positive cirrhosis who had never consumed alcohol (log-rank 0.26, p = 0.61). CONCLUSIONS: Cumulative survival in alcoholic cirrhosis does not seem to be influenced by the presence or absence of markers of HCV infection. Once liver cirrhosis has been diagnosed in the alcoholic patient, complete alcohol abstinence should be strongly recommended. PMID- 12544204 TI - Gastric emptying time and the effect of cisapride in cirrhotic patients with autonomic neuropathy. AB - GOALS To investigate the relationships between gastric emptying and autonomic dysfunction in hepatic cirrhosis and to assess the effects of cisapride on gastric emptying in cirrhotic patients. STUDY Twenty-four cirrhotic patients (8 patients in each Child-Pugh classification) and 25 healthy controls were enrolled. All the patients had viral (B or C) hepatitis. Patients with DM, alcoholic cirrhosis, active peptic ulcer, gastric malignancy and pyloric obstruction were excluded by esophagogastroduodenoscopy. Parasympathetic and sympathetic functions were assessed by the criteria set forth by Ewing and Clark. Drugs affecting GI motility and smoking were discontinued 48 hours and 12 hours prior to the study respectively. A solid-phase of gastric emptying study was conducted by scintigraphic method for the calculation of gastric half-emptying time (GET1/2). RESULTS The study revealed that 9 patients with Child-Pugh B and C cirrhosis had autonomic neuropathy and none of Child-Pugh A cirrhosis had autonomic neuropathy. Prolonged GET1/2 was noted in cirrhotics compared with the control group (p < 0.05). However, there was significant difference between 9 patients (Child B-C) with autonomic neuropathy compared with patients 15 patients without autonomic neuropathy. Again there was a significant difference in GET1/2 between Child A cirrhotic and Child B-C cirrhotic whether they had autonomic neuropathy or not. Cisapride decreased GET1/2 significantly in cirrhotic patients (Child B-C cirrhotic). Clearly, patients with autonomic neuropathy in Child B-C cirrhosis had significantly reduced GET1/2 after cisapride administration. Even though cisapride decreased GET1/2 in patients with Child B-C cirrhosis without autonomic neuropathy, this was not significant. CONCLUSION Autonomic neuropathy in advanced cirrhosis from viral hepatitis may cause prolonged gastric emptying. Cisapride can shorten gastric emptying time in such patients. PMID- 12544205 TI - Chronic graft-versus-host disease complicated by acute hepatitis B. AB - We observed a 45-year-old man with acute hepatitis B while receiving treatment of chronic graft-versus-host disease (GVHD) of the liver. When he developed a sudden elevation of serum aminotransferases 17 months after bone marrow transplantation, he was under immunosuppressive therapy consisting of cyclosporin A against chronic GVHD of the liver. Serologic tests for hepatitis B virus (HBV) showed no reactivation but de novo acute infection. The serum levels of aminotransferases after elevation of biliary tract enzymes increased mildly. However, icterus was not observed in his sequential course. A liver biopsy specimen revealed mild acute liver injury accompanied by slight degeneration of bile ducts. It is presumed that owing to immunosuppressive therapy, his liver dysfunction was relatively mild, and the hepatitis became quiescent without becoming serious. On the other hand, the serum of the patient remained hepatitis B surface antigen positive for more than 1 year after the onset of the hepatitis, which suggested chronicity of HBV infection. PMID- 12544206 TI - Characterization of long-term outcomes after Toupet fundoplication. PMID- 12544207 TI - Reversal of iron deficiency anemia in a patient with gastric antral vascular ectasia treated with cyproheptadine. PMID- 12544208 TI - The incidence of Helicobacter pylori is not increased in obese people, but the risk of duodenal ulcer is higher in H. pylori-positive people of high obesity. PMID- 12544209 TI - Giardia duodenalis: a confounding factor for the diagnosis of celiac disease. PMID- 12544210 TI - Red man syndrome and infliximab. PMID- 12544211 TI - Klebsiella liver abscess. PMID- 12544212 TI - A renal allograft recipient with viral-burden, steatoviral, and fibroviral hepatitis B who achieved remission with Lamivudine. PMID- 12544213 TI - Technology that is driving the systems for the coming decade. PMID- 12544214 TI - A comparison of the stability of single-piece and segmental Le Fort I maxillary advancements. AB - This study retrospectively evaluated the stability of Le Fort I maxillary advancements and compared segmental and one-piece maxillary osteotomy procedures. A cephalometric analysis was performed on 26 cases of maxillary advancement. The sample comprised 11 cases of one-piece and 15 cases of segmental maxillary procedures. The tracings were superimposed and digitized by computer software, and the skeletal changes were analyzed before surgery, immediately after surgery, and at a minimum of 1 year of follow-up. Different values were compared by the paired and nonpaired t tests and were correlated by the Pearson correlation test. The significant value was set at a 95% confidence interval. The maxilla was advanced by a mean of 5.0 +/- 1.6 mm (P < 0.001), and the anterior maxilla was repositioned inferiorly by a mean of 1.5 +/- 3.3 mm (P < 0.05). The maxilla relapsed posteriorly by a mean of 0.6 +/- 1.2 mm (P < 0.05) and superiorly at the anterior maxilla by a mean of 0.8 +/- 1.1 mm (P < 0.001). Overjet and overbite did not significantly change (P > 0.05). It was concluded that maxillary advancement using rigid fixation and interpositional bone grafting in both groups was a stable procedure, particularly in the horizontal plane. In the one-piece group, there was a significantly higher relapse in the vertical plane than in the segmental group (P < 0.05), however. Minor skeletal relapse was compensated for by postoperative tooth movement, and segmental procedures are recommended when required to enhance occlusal results. PMID- 12544215 TI - Anatomical variations of the frontal and supraorbital transcranial passages. AB - In this study, 500 frontal and supraorbital transcranial passages were studied in 50 cadavers and 200 crania of 500 samples. One hundred six specimens had a frontal foramen (notch), and all the samples had a supraorbital foramen (notch). The frontal passage was a foramen in 7 samples and a notch in 99 samples. As for the supraorbital passage, it was found as a foramen in 133 of specimens and as a notch in 358 of specimens. In 9 of the specimens, there were double notches or foramina. The distances from the foramina (notches) to angulus oculi medialis were measured in the cadavers. The average distance from the angulus oculi medialis to the frontal foramen (notch) was 4.50 mm, and the average distance to the supraorbital foramen (notch) was 9.87 mm. The distances from the foramina (notches) to the midline were measured in the crania. The average distances from the midline to the frontal foramen (notch) and the supraorbital foramen (notch) were 20.24 mm and 25.23 mm, respectively. The average distance between the frontal foramen (notch) and supraorbital foramen (notch) was 5.37 mm in cadavers and 4.99 mm in crania. In 200 crania, the distances of the frontal and supraorbital transcranial passages to the midline were measured. Types of these passages were also evaluated, and frequencies were calculated. Measurements were made using a digital compass, and the student t test was used in the statistical evaluation of results. PMID- 12544216 TI - Proportion indices in the craniofacial regions of 284 healthy North American white children between 1 and 5 years of age. AB - The study's aim was to provide the normal range data on 81 anthropometric proportion indices of the craniofacial complex, separately in boys and girls between 1 and 5 years of age, supplementary to the previously published normal proportion values of the North American white population from 6 to 18 years of age. In the first age group, 17 or 18 boys and 18 or 22 girls were measured. In each age group between 2 and 5 years old, 30 boys and 30 or 31 girls were examined. The normal values of the 81 proportion indices were established with the help of 45 measurements (7 cranial, 13 facial, 4 orbital, 12 nasal, 7 labio oral, and 2 auricular) taken from 287 healthy children. The data on the normal indices were reported in two groups: areal and interareal. The areal proportions are formed by taking measurements from one particular craniofacial region. Proportions indicating the quality of relation of measurements taken from two craniofacial regions were classified as interareal (N = 36). The basic proportions (N = 13) of the areal one show the relation between the measurements influencing the general view of each craniofacial region. The detailed analysis of the proportionality in the individual regions, carried out with the help of the other measurements within the regions, established 36 additional areal proportion indices. The recommended method of examination is to start with the basic proportions, which offers the determination of the most and least disproportionate craniofacial region, followed by a more comprehensive search for other disproportions with the help of the additional areal proportions (N = 36) and interareal proportions (N = 32). PMID- 12544217 TI - Three-dimensional cranial expansion using distraction osteogenesis for oxycephaly. AB - Oxycephaly is associated with raised intracranial pressure as a result of the fusion of multiple cranial sutures. We have performed an effective and less invasive cranial expansion by means of three-dimensional cranial distraction for the treatment of oxycephaly with suspicion of increased intracranial pressure. We describe two oxycephaly cases and the surgical technique of three-dimensional cranial expansion using distraction osteogenesis. PMID- 12544218 TI - Acute bacterial suppurative parotitis: microbiology and management. AB - The parotid gland is the salivary gland most commonly affected by inflammation. The most common pathogens associated with acute bacterial parotitis are Staphylococcus aureus and anaerobic bacteria. The predominant anaerobes include gram-negative bacilli (including pigmented Prevotella and Porphyromonas spp.), Fusobacterium spp., and Peptostreptococcus spp. Streptococcus spp. (including S. pneumoniae) and gram-negative bacilli (including Escherichia coli) have also been reported. Gram-negative organisms are often seen in hospitalized patients. Organisms less frequently found are Arachnia, Haemophilus influenzae, Klebsiella pneumoniae, Salmonella spp., Pseudomonas aeruginosa, Treponema pallidum, cat scratch bacillus, and Eikenella corrodens. Mycobacterium tuberculosis and atypical mycobacteria are rare causes of parotitis. Therapy includes maintenance of hydration and administration of parenteral antimicrobial therapy. Once an abscess has formed surgical drainage is required. The choice of antimicrobial depends on the etiologic agent. Maintenance of good oral hygiene, adequate hydration, and early and proper therapy of bacterial infection of the oropharynx may reduce the occurrence of suppurative parotitis. PMID- 12544219 TI - An unusual case of heminasal aplasia: proboscis lateralis or cystic teratoma? AB - Unilateral aplasia of the nose is a rare congenital malformation. It is often associated with other malformations of the facial region, including abnormalities of the eye and lacrimal system, proboscis lateralis, and facial bone malformations. The authors present a case of heminasal aplasia in a 4-year-old boy who underwent surgery immediately after birth because of right-sided proboscis lateralis. A huge hemispheric tumefaction was occupying a large part of the right orbit, medial canthus, and frontonasal region of the face, causing lateroinferior protrusion of the eyeball. The right half of the external nose was aplastic. The patient was operated on by an external approach. The pathohistological findings from the top of tumefaction revealed cystic teratoma. The possible backgrounds for the onset of the tumefaction are discussed in two main directions: the cystic form of the regrowth of the remnants of inadequately excised proboscis lateralis and cystic teratoma of the ethmoidal sinus. PMID- 12544220 TI - False diagnosis caused by Warthin tumor of the parotid gland combined with actinomycosis. AB - A case is reported in which a unilateral parotid gland cystadenolymphoma was combined with actinomycosis. A 48-year-old woman presented with a mass in the left parotid region and paresis of the lower left palpebra. The computed tomography, echography, and parotid radiographic findings did not exclude a neoplasm of the left parotid gland. The ramus of the mandible was involved in the process. Intraoperative freezing histology, total parotidectomy, and partial mandibulectomy were performed, with sacrifice of the facial nerve followed by nerve reconstruction. The final histological evaluation was Warthin tumor with actinomycosis. Four years after treatment, the patient is free of disease. No similar cases seem to have been reported thus far. PMID- 12544221 TI - Diagnosis of Bell palsy with gadolinium magnetic resonance imaging. AB - Bell palsy is a condition resulting from a peripheral edematous compression on the nervous fibers of the facial nerve. This pathological condition often has clinical characteristics of no importance and spontaneously disappears in a short time in a high percentage of cases. Facial palsy concerning cranial nerve VII can also be caused by other conditions such as mastoid fracture, acoustic neurinoma, tumor spread to the temporal lobe (e.g., cholesteatoma), neoformation of the parotid gland, Melkersson-Rosenthal syndrome, and Ramsay-Hunt syndrome. Therefore, it is important to adopt an accurate diagnostic technique allowing the rapid detection of Bell palsy and the exclusion of causes of facial paralysis requiring surgical treatment. Magnetic resonance imaging (MRI) with medium contrast of the skull shows a marked increase in revealing lesions, even of small dimensions, inside the temporal bone and at the cerebellopontine angle. The authors present a clinical case to show the important role played by gadolinium MRI in reaching a diagnosis of Bell palsy in the differential diagnosis of the various conditions that determine paralysis of the facial nerve and in selecting the most suitable treatment or surgery to be adopted. PMID- 12544222 TI - Orthodontic-surgical treatment after posttraumatic bilateral condylectomy of the mandible in an adult patient. AB - A posttraumatic open bite associated with a bird face is reported. Condylectomy was indicated in relation to the plurifragmentary fracture of the condyles with limitation of mandibular movement. Condylectomies were mandatory as a result of delayed maxillofacial surgical treatment, which was related to the poor general condition of the patient after trauma. Functional disorder recovery and aesthetic deformity correction were planned by a team approach between orthodontists and maxillofacial surgeons with the support of a logopedist for the postural-related muscle problems. The main practical and theoretical problems presented by the clinical case were a result of the need to restore the occlusal relations and to avoid recurrence of open bite in this patient. The patient presented a wide alteration of muscular function and a strength fibrotic retraction with alteration in the relationship between upper and lower jaws and retrusion of the mandible associated to open bite. Orthodontic treatment was carried with no impact on the upper and lower axis, avoiding orthodontic correction of the open bite. Surgery corrected both the open bite and the bird face by means of bilateral sagittal split osteotomies. Wiring of the mandibular osteotomies and intermaxillary fixation allowed positioning of the mandibular ramus bilaterally because of the fibrosis and muscular action-related forces without resulting in a similar rotation of the mandible with the risk of recurrence. Myotherapy and logopedic support minimized the risk of recurrence, improved reduction of muscular tension with the resolution of the lip incompetence, and allowed functional recovery of mandibular movements. PMID- 12544223 TI - Porous polymethylmethacrylate as bone substitute in the craniofacial area. AB - In craniofacial surgery, alloplastic materials are used for correcting bony defects. Porous polymethylmethacrylate (PMMA) is a biocompatible and nondegradable bone cement. Porous PMMA is formed by the classic bone cement formulation of methylmethacrylate liquid and PMMA powder in which an aqueous biodegradable carboxymethylcellulose gel is dispersed to create pores in the cement when cured. Pores give bone the opportunity to grow in, resulting in a better fixation of the prostheses. We evaluated the long-term results (n = 14), up to 20 years, of augmentations and defect fillings in the craniofacial area, with special interest in possible side effects and bone ingrowth. The evaluation consisted of a questionnaire, a physical examination, and a computed tomography (CT) scan. There were no side effects that could be ascribed to the porous PMMA. Twelve CT scans showed bone ingrowth into the prostheses, proving the validity behind the concept of porous PMMA. PMID- 12544224 TI - Brown tumor in children with normocalcemic hyperparathyroidism: a report of two cases. AB - Brown tumor is an important sign of a number of metabolic bone diseases. Normocalcemic hyperparathyroidism, a rare childhood pathologic finding, reveals itself with brown tumors as the only initial sign. In this study, two cases of normocalcemic hyperparathyroidism accompanied by mandibular brown tumors are presented. PMID- 12544225 TI - Malignant fibrous histiocytoma of scalp with intracranial invasion 20 years after postoperative irradiation. AB - We report a case of malignant fibrous histiocytoma with intracranial invasion occurring 20 years after radiation therapy for astrocytoma. Magnetic resonance imaging suggested that a dumbbell-shaped tumor arising from soft tissue was pressing the brain. Histopathological findings revealed a storiform pattern with spindle cells such as fibroblasts, histiocytes, epithelioid cells with irregularly shaped nuclei, and bizarre giant cells. Three operations involving craniectomy were necessary because of recurrence. PMID- 12544226 TI - Etiology and patterns of facial fractures in the United Arab Emirates. AB - A retrospective study on facial fractures was carried out in the Department of Oral and Maxillofacial Surgery at Tawam Hospital (Al Ain, United Arab Emirates) between January 1, 1998 and December 31, 2001. The study included 144 patients with a mean age of 26.5 years; the most frequently injured patients belonged to the 16- to 20-year-old age group. The male predilection was 83%. Road traffic accident was the most common causative factor (59%), followed by falls (21.5%), accidents where camels were involved (5.5%), work- and sport-related accidents (4.8% and 4.8%, respectively), and assault (4.1%). A total of 53.4% of the patients suffered isolated mandibular fractures, 32.6% had isolated midface fractures, and 13.8% had combined midface and mandibular fractures. Associated injuries were noted in 22.2% of the patients. The number of patients treated increased from 28.3 (1990-1995) to 36 (1998-2001) on an annual average; a reduction in isolated nasal fractures and associated injuries, including facial lacerations, was noted with no change in age or etiology predilection. Sufficient data could not be obtained to determine if the favorable results with the associated injuries were a result of the effect of a compulsory seat belt law introduced on June 1, 1998, but the increasing number of maxillofacial injuries suggests that the seat belt law is ignored in this country. PMID- 12544227 TI - Hydroxyapatite cranioplasty: I. Experimental results from a new quick-setting material. AB - The biophysical characteristics of a new formulation of hydroxyapatite (HA) for craniofacial applications having a rapid set time was evaluated in animals. Using a mature rabbit model, the material was applied in both cranial defects and as a geometrically shaped cranial onlay. Visual and histological assessment was done after 1 year of implantation. The material remained morphologically stable after 1 year, with complete preservation of its onlay shape and stable fill of the cranial defects. A significant osteoconductive response was seen with bone growth across the inner and outer surfaces of the inlays and along the sides of the onlays. No fibrovascular ingrowth or bony replacement of the material itself was evident as a result of the age of the study animals or the compact density of the material. This rapid-setting HA formulation offers similar favorable tissue responses as those of other longer setting HA materials. PMID- 12544228 TI - Long-spanning resorbable plates in cranial vault reconstruction. PMID- 12544229 TI - A new technique for correction of trigonocephaly using bioabsorbable osteofixation tacks and plates and a novel tack-shooter. AB - We report on the feasibility of applying bioabsorbable tacks using a new tack shooter to fix bioabsorbable plates applied endocranially for the correction of three cases of trigonocephaly. Tacks do not require tapping or tightening because they are applied using a tack-shooter directly into drill holes in the bone. Hence, the technique saves valuable operative time. A 1.5- to 2.0-cm broad supraorbital bar (bandeau) was raised and reshaped. The corrected shape was maintained using a Biosorb plate (Bionx Implants Ltd, Tampere, Finland), and tacks were applied on the endocranial side of the bar. The plate extended a few centimeters laterally beyond the edge of the supraorbital bar, and it was fixed with Biosorb miniscrews and/or tacks affixed to the temporal bones. Other molded bone pieces were fixed using Biosorb plates, screws, and/or tacks. The technique of using tacks was easy, and it provided secure osteofixation. Cosmetic results were excellent, and no complications were encountered except for palpability of plate edges on the right side of the skull in one case. PMID- 12544230 TI - Large osteomas of the cranial vault. PMID- 12544231 TI - FGFR2 mutations among Thai children with Crouzon and Apert syndromes. AB - Crouzon and Apert syndromes have been reported to be associated with mutations in Fibroblast Growth Factor Receptor 2 (FGFR2) gene in various ethnic groups, but never in Southeast Asian subjects. Therefore, the authors conducted a study to characterize 11 Thai patients: four with Crouzon syndrome and seven with Apert syndrome. All cases are sporadic. Mean paternal and maternal ages were 38.7 and 28.6 years, respectively. Molecularly, all patients were found to have mutations in the FGFR2 gene. Three mutations (C278F, S347C, S351C) were detected in all Crouzon patients with two having S351C. The seven patients with Apert syndrome have either S252W or P253R mutation. The authors' findings that sporadic cases were associated with advanced paternal age and that they all had mutations in FGFR2 are consistent with previous reports. This is another observation supporting the causative role of FGFR2 mutations in Crouzon and Apert syndromes. PMID- 12544232 TI - Experimental study on bone formation potential of cryopreserved human bone marrow mesenchymal cell/hydroxyapatite complex in the presence of recombinant human bone morphogenetic protein-2. AB - Secondary bone grafting in the alveolar cleft is one of the most important treatment modalities for patients with a cleft lip and palate. In children, however, harvesting a sufficient amount of bone graft from the donor site is difficult, and the procedure imposes a heavy burden on the patients. This problem may be resolved if autologous transplantation can be performed using a cell hybrid type of artificial bone prepared with the patient's own bone marrow cells through a tissue engineering technique. In the current study, we examined the possibility of achieving such an autologous transplantation using cryopreserved human bone marrow cells. Human bone marrow cells were grown in culture and cryopreserved, and the cells preserved for 3 years and the cells preserved for 3 months were then thawed and recultivated. In one experiment, the recultivated cells were seeded onto a complex composed of porous hydroxyapatite and bone morphogenetic protein (BMP), and the complex was grafted subcutaneously in nude mice. In another experiment, the cells were seeded onto the porous hydroxyapatite and cultivated for 10 days before grafting in a medium supplemented with BMP. The bone formation potential of the cells was compared between these two experiments. Formation of mature bone was observed 2 weeks after transplantation in the former experiment, whereas only scarce bone formation was evident 4 weeks after transplantation in the latter experiment. It is therefore assumed that exposing the cultured human bone marrow cells to a high concentration of BMP in vivo strongly promotes bone formation. PMID- 12544233 TI - Reconstruction of the immature craniofacial skeleton with a carbonated calcium phosphate bone cement: interaction with bioresorbable mesh. AB - Calcium phosphate cements have been recently introduced for use in craniofacial reconstruction. In the clinical setting, however, pulsations of the underlying brain and dura may interfere with the crystallization of these cements, thereby rendering their use in cranioplasty problematic. To circumvent such problems, many clinicians have interposed synthetic resorbable plates or mesh between the dura and the cement. At the present time, however, little is known about the influence of such materials or their breakdown products on the fate of calcium phosphate cements. The specific aim of this project was to evaluate the biocompatibility, osteoconductivity, and remodeling capacity of a calcium phosphate cement after implantation into experimental calvarial defects when combined with a resorbable mesh underlay. Four 10-mm diameter full-thickness calvarial defects (two frontal, two parietal) were created in each of six 3-week old Yorkshire pigs. The defects were treated as follows: 1) empty control, 2) macroporous polylactic acid (70/30 L/DL polylactic acid [PLA]) mesh, 3) Norian CRS calcium phosphate cement, and 4) Norian CRS over PLA mesh underlay. Animals were divided into two groups. Half of the animals were killed 30 days after surgery, and half were killed 180 days after surgery, and the graft recipient sites were examined histologically. At 30 days, minimal bone ingrowth was observed in untreated calvarial defects or in those that were treated with PLA plates alone. Defects treated with the cement alone demonstrated a modest amount of new woven bone deposition, primarily at the periphery of the implants. Defects treated with calcium phosphate cement over PLA mesh underlays were characterized by remodeling and woven bone deposition at 30 days, with complete or near complete osseous bridging of the ectocranial implant surfaces. Progressive bone ingrowth was noted in all defects at 180 days, with near-complete replacement of all Norian CRS implants by host bone. The PLA mesh remained incompletely resorbed at 180 days. No inflammatory response to the implants was observed at either time point. Calcium phosphate cement may be safely used for craniofacial reconstruction in the presence of PLA implants without compromise to its biocompatibility, osteoconductivity, or remodeling capacity. PMID- 12544235 TI - Diffusion-weighted imaging for the evaluation of diffuse axonal injury in closed head injury. AB - PURPOSE: The purpose of this work was to compare diffusion-weighted imaging (DWI) with conventional MRI in the detection of shearing injuries in acute closed head injuries. METHOD: Twenty-five patients (19 male, 6 female) were examined within 48 h of trauma. Conventional MRI included T2-weighted fast spin echo, fluid attenuated inversion recovery (FLAIR), and T2*-weighted gradient echo sequences. Full tensor DWI with calculation of apparent diffusion coefficient (ADC) maps was also performed. Lesions were identified and compared on all sequences. RESULTS: Four hundred twenty-seven lesions were counted by the combined use of all sequences. DWI identified 70 lesions not seen on conventional MRI. DWI identified 310 shearing injuries, followed by T2/FLAIR (n = 248) and T2* (n = 202). The majority of DWI-positive lesions showed decreased diffusion (65%). CONCLUSION: DWI is valuable in closed head injury because it identifies additional shearing injuries not visible on T2/FLAIR or T2* sequences. Furthermore, DWI/ADC maps differentiate between lesions with decreased or increased diffusion. DWI is less sensitive than T2* imaging for detecting hemorrhagic lesions. PMID- 12544236 TI - Central nervous system paracoccidioidomycosis: imaging findings in 17 cases. AB - PURPOSE: To quantify the imaging findings of 17 patients with central nervous system (CNS) paracoccidioidomycosis. METHOD: Retrospective analysis of the clinical data, films, and laboratory findings of 17 patients with CNS paracoccidioidomycosis. The clinical data were obtained by medical records review. Plain chest films, cranial computed tomography scans (CT), and serum and liquor analysis were evaluated. RESULTS: All the patients were male, with a mean age of 46.5 years. Headache and hemiparesis were the most frequent symptoms. The head CT scans revealed two or more granulomas in 53% of the cases, frequently confined to the parietal lobes (35%) and cerebellar hemispheres (35%). The lesions were hypodense (53%) and irregular (76%) and showed ring-like contrast enhancement (94%). Perilesional edema was seen in 82% of the patients, and hydrocephalus was seen in 41%. CONCLUSION: Although CNS paracoccidioidomycosis might represent a suspicious lesion pattern on CT scan examination, correlation of the CT scan findings, clinical data, plain chest films, and laboratory results is necessary to define the diagnosis of this uncommon entity. PMID- 12544237 TI - Esophageal invasion by thyroid carcinomas: prediction using magnetic resonance imaging. AB - PURPOSE: We evaluated the accuracy of magnetic resonance imaging (MRI) in predicting esophageal invasion by thyroid carcinomas and established an optimal criterion for diagnosing esophageal invasion. METHOD: The MRI findings (size and margins of tumor, ratio of tumor contact to the esophagus, shape and displacement of the esophagus, and tumor invasion to the outer and inner layers of the esophagus) in 67 patients with thyroid carcinomas were retrospectively reviewed and correlated with surgical and pathologic findings. Logistic modeling was used to determine the significant factors for predicting esophageal invasion. RESULTS: Seventeen (34%) of the 67 patients had pathologically or surgically verified esophageal invasion. The logistic modeling revealed that outer layer invasion (P < 0.001) and poorly defined margins (P = 0.001) were the significant factors. The outer layer invasion showed the highest accuracy of 91%, with 82% sensitivity and 94% specificity. The addition of poorly defined margins to this criterion did not improve its accuracy. CONCLUSION: Esophageal invasion by thyroid carcinoma was accurately predicted with MRI, and an MRI finding of outer layer invasion was optimal for diagnosing esophageal invasion. PMID- 12544238 TI - Neuroimaging findings in a suprasellar granular cell tumor. AB - A 27-year-old woman presented with a history of amenorrhea, visual disturbance, and diabetes insipidus. Magnetic resonance imaging showed a large enhancing suprasellar mass with associated edema involving the left striatum. The lesion was hypometabolic on fluorodeoxyglucose-positron emission tomography. At surgery, a subtotal resection of a vascular tumor that appeared to arise from the posterior pituitary and hypothalamus was carried out. Pathologic examination revealed a granular cell tumor. We report the preoperative neuroimaging findings in this rare posterior pituitary stalk tumor. PMID- 12544239 TI - Canavan disease: diffusion magnetic resonance imaging findings. AB - A 15-month-old boy with Canavan disease is reported in whom a restricted diffusion pattern on diffusion magnetic resonance imaging (MRI) (high signal on b = 1,000 mm2/s images and low apparent diffusion coefficient [ADC] values) was evident in the affected regions of the brain, including the peripheral white matter, globi pallidi, thalami, brainstem, dorsal pons, and dentate nuclei. The ADC values at these regions ranged from 0.42 to 0.56 x 10(-3) mm2/s compared with the normal ADC values from the uninvolved deep frontal white matter (0.68-0.92 x 10(-3) mm2/s). The known histopathologic features in Canavan disease include edematous and gelatinous brain tissue associated with diffuse vacuolization. Considering these and the diffusion MRI findings in this patient, it is likely that existence of a gel (gelatinous) state rather than the usual sol state of water molecules in the affected brain regions accounted for the restricted diffusion pattern in Canavan disease. PMID- 12544240 TI - Diffusion magnetic resonance imaging in infantile neuroaxonal dystrophy. AB - A 7-month-old girl with infantile neuroaxonal dystrophy is reported. In diffusion MRI, the pyramidal tracts and dentate nuclei had high signal on b = 1,000 s/mm2 images and low apparent diffusion coefficient (ADC) values. This pattern likely reflected the presence of abnormal (dystrophic) axons with restricted mobility of water molecules. A reverse pattern was evident in the cerebellar cortex with high ADC values. This was likely a reflection of dysmyelination or lack of myelination. PMID- 12544241 TI - L-2 hydroxyglutaric aciduria: proton magnetic resonance spectroscopy and diffusion magnetic resonance imaging findings. AB - A 10-month-old boy was reported with the diagnosis of L-2 hydroxyglutaric aciduria. Amino acid chromatographic analysis revealed an 80-fold increase of hydroxyglutaric acid in the urine. Proton magnetic resonance (MR) spectroscopy of the brain obtained with the hybrid chemical shift imaging sequence (repetition time = 1,500 milliseconds, echo time = 40 milliseconds) revealed prominent peaks resonating at 2.50 ppm, which were attributable to L-2 hydroxyglutaric acid. Diffusion MR imaging was obtained using the echo-planar trace sequence (repetition time = 5,700 milliseconds, echo time = 139 milliseconds). Two different diffusion patterns were evident: a restricted diffusion pattern in the globi pallidi and an increased diffusion pattern in the white matter. PMID- 12544242 TI - Magnetic resonance spectroscopy in a 9-day-old heterozygous female child with creatine transporter deficiency. AB - An X-linked creatine deficiency syndrome caused by mutations in the creatine transporter gene SLC6A8/CRTR mapped to Xq28 has recently been described. Essential in the recognition of this disorder is the absence of creatine on proton magnetic resonance spectroscopy (MRS) examination. A 9-day-old heterozygous female child with this syndrome demonstrated a significant reduction of creatine on proton MRS. She is a carrier of the R514X nonsense mutation. PMID- 12544243 TI - Accuracy in differentiation of mucinous and nonmucinous rectal carcinoma on MR imaging. AB - PURPOSE: The purpose of this work was to determine the diagnostic accuracy of MRI for the differentiation of mucinous and nonmucinous carcinomas in patients with rectal carcinomas. METHOD: Seventy-six patients with pathologically proven rectal adenocarcinomas were evaluated with MRI. Both T2-weighted fast spin echo and T1 weighted spin echo images were independently reviewed by two radiologists who were blinded for tumor histology. Of the 76 cases, mucin pools were found in 28 (37%), and 16 had a mucin pool occupying >or=50% of the tumor (mucinous carcinoma). The sensitivity, specificity, accuracy, and positive and negative predictive values of distinguishing between mucinous and nonmucinous carcinomas were calculated. The interobserver agreement was estimated using kappa statistics. RESULTS: The accuracies of the diagnosis of mucinous carcinoma were 97% for Radiologist 1 and 96% for Radiologist 2. The sensitivities and specificities for diagnosing a mucinous carcinoma were 94 and 98% for Radiologist 1 and 100 and 95% for Radiologist 2, respectively. Interobserver agreement was good (kappa = 0.61) for the identification of a mucin pool and was excellent (kappa = 0.81) for the diagnosis of a mucinous carcinoma. CONCLUSION: Differentiation of mucinous and nonmucinous rectal carcinoma is highly accurate on preoperative MRI. PMID- 12544244 TI - Fractal analysis of internal and peripheral textures of small peripheral bronchogenic carcinomas in thin-section computed tomography: comparison of bronchioloalveolar cell carcinomas with nonbronchioloalveolar cell carcinomas. AB - PURPOSE: To analyze the internal and peripheral textures of small peripheral bronchogenic carcinomas (<2 cm) in thin-section computed tomography (HRCT) images with fractal analysis. METHOD: Thin-section computed tomography images from 70 patients with bronchogenic carcinomas (61 adenocarcinomas and 9 squamous cell carcinomas) were used. Regions of interest (ROIs) with a matrix size of 32 x 32 (0.326 mm per pixel) were selected manually on the lung-nodule interfaces and within the nodules on HRCT images. Three-dimensional density surfaces based on CT values of ROIs were characterized by fractal dimensions (FDs). RESULTS: When all the bronchogenic carcinomas were divided into bronchioloalveolar cell carcinomas (BACs) and other bronchogenic carcinomas (nonBACs), there were significant differences between BACs and nonBACs in the FDs obtained from the internal textures (mean: 2.38 +/- 0.05 versus 2.19 +/- 0.05; P< 0.0001) and in the FDs obtained from the peripheral textures (mean: 2.16 +/- 0.01 versus 2.06 +/- 0.01; P< 0.0001). CONCLUSION: The textures of BACs that reveal ground-glass opacities are more complicated than those of nonBACs. The FDs can differentiate between small localized BACs, which have a good prognosis, and nonBACs, which have a poor prognosis. Fractal analysis is promising for characterization of small peripheral pulmonary bronchogenic carcinomas based on radiographic features of HRCT images. PMID- 12544245 TI - Image quality in a standardized algorithm for minimally invasive coronary angiography with multislice spiral computed tomography. AB - PURPOSE: To report our experience with a standardized approach to pharmacologic heart rate control and image postprocessing for computed tomographic coronary angiography (CTCA) with multislice computed tomography (MSCT). METHOD: Two experienced observers used transaxial tomograms and maximum-intensity projections to classify coronary segments (12 per patient, 135 consecutive patients) for degree of stenosis. One factor affecting image quality was identified for each segment that could not be assessed. RESULTS: Nine patients (7%) were excluded for technical reasons. Of 1,512 segments from 126 patients, 1,086 (72%) were assessable (8.6 per patient). Of 300 segments from 25 patients who also had selective coronary angiography, CTCA was able to assess 211 (70%) and detected significant disease in 27 (82% sensitivity, 96% specificity, 73% positive predictive value, and 97% negative predictive value). Vessel caliber, heart rate, and Agatston score were associated with inability to assess 426 coronary segments (28%). CONCLUSION: Heart rate and Agatston score are important predictors of the ability to assess proximal and midcoronary segments by CTCA with MSCT. PMID- 12544246 TI - Evaluation of hemangioma by positron emission tomography: role in a multimodality approach. AB - PURPOSE: The relative utility of various preoperative diagnostic imaging modalities for the evaluation of hemangioma of the extremities, including positron emission tomography (PET) (using 18F-fluoro-2-deoxy-D-glucose [FDG] and fluorine-18 alpha-methyltyrosine [FMT]), computed tomography (CT), magnetic resonance imaging (MRI), and digital subtraction angiography (DSA), was investigated. METHODS: Imaging findings in 16 patients with 16 histopathologically documented hemangiomas of the extremities were retrospectively reviewed. Preoperative imaging included: FDG-PET (n = 16), FMT PET (n = 12), MRI (n =16), CT (n =11), and DSA (n =14). RESULTS: All 16 lesions examined by PET with FDG and/or FMT showed accumulation. The standardized uptake values (SUVs) for FDG-PET for the 16 examined tumors ranged from 0.7 to 1.67; for FMT-PET, they ranged from 0.14 to 1.00. The SUVs with both tracers indicated the benign nature of the tumor. Computed tomography demonstrated variable attenuation and phleboliths in two patients. The MRI signal characteristics were relatively consistent: heterogeneous signals were slightly higher than those of skeletal muscle on T1-weighted images and brighter than those of subcutaneous fat on T2 weighted images. The pooling and cotton-wool staining depicted in DSA was found to be significantly correlated with FDG accumulation, suggesting that localized blood retention-induced ischemia may accelerate anaerobic glycolysis, which leads to high FDG uptake. CONCLUSION: Although plain radiography, CT, MRI, and angiography may provide anatomic extent and be pathognomonic, FDG-PET and FMT-PET may be the most reliable among the studied imaging modalities for differentiating benign hemangiomas from other soft tissue tumors, especially malignant neoplasms. PMID- 12544247 TI - Usefulness of electron beam tomography in the prone position for detecting atrial thrombi in chronic atrial fibrillation. AB - OBJECTIVE: We compared the usefulness of electron beam tomography (EBT) in the prone position relative to that in the supine position for detecting atrial thrombi. METHODS: We studied 96 patients with chronic atrial fibrillation, of whom 71 were scanned in the supine position and 25 were scanned in the prone position. Electron beam tomography was performed twice after contrast medium injection to obtain early- and late-phase images. RESULTS: Filling defects were detected in 13 patients in the supine position by EBT. Transesophageal echocardiography (TEE) revealed a thrombus in the region of the filling defect in 9 patients. In 4 patients, filling defects in the left atrial appendage were not confirmed as thrombi by TEE. Filling defects were detected in 4 patients in the prone position, all of which were confirmed as thrombi by TEE. CONCLUSION: Electron beam tomography in the prone position is an effective technique for reducing false-positive results in the detection of atrial thrombi. PMID- 12544248 TI - Lithiasis in a heterotopic pancreas of the stomach. AB - A heterotopic pancreas is a relatively uncommon abnormality and usually of no clinical importance. A heterotopic pancreas is subject to various pathologic changes occurring in the pancreas, however, such as acute pancreatitis, cyst or abscess formation, pancreatic cancer, and islet cell tumor. We describe a rare case of a heterotopic pancreas presenting as a gastric submucosal mass with punctate calcifications occurring in a 43-year-old man. PMID- 12544249 TI - Retained products of conception masquerading as acquired arteriovenous malformation. AB - This case documents a seldom-described event of retained products of conception masquerading as an acquired arteriovenous malformation (AVM) of the uterus. The patient presented with sudden onset of heavy vaginal bleeding 6 weeks after artificial abortion. Ultrasonography and magnetic resonance imaging revealed a diffuse intramural lesion predominantly consisting of prominent vessels, which raised suspicion of an AVM. Hysterectomy revealed retained products of placenta that were necrotic and encroached into thin myometrium. PMID- 12544250 TI - Sudden cardiac arrest during computed tomography examination: clinical findings and "dense abdominal veins" on computed tomography. AB - We report imaging findings in five patients who had sudden cardiac arrest during contrast-enhanced computed tomography (CT). We observed strikingly dense abdominal veins, variable degrees of arterial enhancement, and poor abdominal visceral enhancement. Comparison with a control group of 30 patients revealed a statistically significant increase in mean enhancement of the abdominal veins (including the inferior vena cava, bilateral renal veins, and major tributaries of the hepatic vein) (Kruskal-Wallis test, P< 0.05). These patients lacked any clear cause of cardiac arrest, and the arrest may have been related to an adverse reaction to contrast medium. Despite resuscitation, two patients died and three convalesced. In summary, sudden cardiac arrest is characterized by "dense abdominal veins" on CT in the absence of the cardiac pump function. This effect was presumably the result of forced reflux and stagnation of contrast medium in the abdominal veins without any dilution with the circulating blood, leading to a markedly dense appearance of these venous structures. PMID- 12544251 TI - Adult cranial dura II: venous sinuses and their extrameningeal contributions. AB - This is the second of two articles designed to provide detailed user-friendly schematics of the adult dural vascular anatomy. It supplements the first article on the intrinsic meningeal arteries and veins by describing the dural venous sinuses along with their three remaining connections: to the brain, to the diploe, and to the extracranial soft tissues. The discussion of this anatomy is supplemented by illustrative pathologic insights. This information will assist in interpreting neuroimaging studies, communication with clinicians, and teaching of this complex subject. PMID- 12544252 TI - Efp as a new molecular target for breast cancer therapy. PMID- 12544253 TI - Inducing cancer cell death by targeting transcription factors. AB - We review the biological significance of transcription factors such as p53, Myc, E2F family and AP-1 (Jun/Fos) in anticancer drug-induced apoptosis. It is likely that the functional role of these transcription factors is complex in response to DNA damage depending on cancer cell type. Regulation of apoptosis following DNA damage is mediated by cell cycle arrest for DNA repair and subsequent signal transduction pathways leading to apoptosis, which is associated with mitochondrial dysfunction. Activation of transcription factors following anticancer drugs is located upstream of signal transduction pathways, thereby the downstream pathway is promoted, which is connected to activation or suppression of apoptosis-related proteins. Switching on apoptotic signals by anticancer drugs is amplified in mitochondria by releasing cytochrome from the ion channel to activate the caspase cascade, which is regulated by Bcl-2 families in the central gate for drug-induced apoptosis. Activation of transcription factors targeting downstream genes, some of which are apoptosis-related genes, can play a critical role in promoting apoptosis following treatment with anticancer drugs. The strategy of identification of downstream target proteins or transcription factors involved in apoptosis will be necessary for the development of an effective transcription factor-targeted chemotherapy for cancer. PMID- 12544254 TI - Paclitaxel at ultra low concentrations inhibits angiogenesis without affecting cellular microtubule assembly. AB - Many conventional chemotherapeutics, such as the microtubule-stabilizing anticancer drug paclitaxel (Taxol), have been shown to have anti-angiogenic activity and clinical application of a continuous low dose of these agents has been suggested for cancer therapy. In this study, we show that paclitaxel selectively inhibits the proliferation of human endothelial cells (ECs) at ultra low concentrations (0.1-100 pM), with an IC50 = 0.1 pM, while it inhibits non endothelial type human cells at 10(4) - to 10(5) -fold higher concentrations, with IC50 = 1-10 nM. The selectivity of paclitaxel inhibition of cell proliferation is also species specific, as mouse ECs are not sensitive to paclitaxel at ultra low concentrations. They are inhibited by higher concentrations of paclitaxel with IC50 = 1-10 nM. Inhibition of human ECs by paclitaxel at ultra low concentrations does not affect the cellular microtubule structure, and the treated cells do not show G2/M cell cycle arrest and apoptosis, suggesting a novel but as yet unidentified mechanism of action. In an in vitro angiogenesis assay, paclitaxel at ultra low concentrations blocks human ECs from forming sprouts and tubes in the three-dimensional fibrin matrix. In summary, paclitaxel selectively inhibits human EC proliferation and in vitro angiogenesis at low picomolar concentrations. The data support a clinical application of continuous ultra-low-dose paclitaxel to treat cancer. PMID- 12544255 TI - In vitro studies on the lymphoma growth-inhibitory activity of sulfasalazine. AB - Sulfasalazine (SASP) is a novel, potent inhibitor of cellular cystine uptake mediated by the x(c)- cystine/glutamate antiporter. Lymphoid cells cannot synthesize cyst(e)ine and depend for growth on its uptake from their micro environment. We previously showed that SASP (0.2 mM) can abrogate lymphoma cell proliferation in vitro by specifically inhibiting x(c)- -mediated cystine uptake. Intraperitoneal administration of SASP to Noble rats markedly suppressed Nb2-U17 rat lymphoma transplant growth, notably without major toxicity to the hosts. Since Nb2-U17 cells are x(c)- -deficient, the growth arrest was apparently not due to SASP-tumor cell interaction, but possibly to interference with x(c)- mediated cysteine secretion by somatic cells. In this study we found that replication of x(c)- -deficient Nb2-11 lymphoma cells can be sustained in vitro, in the absence of cystine uptake enhancers, by co-culturing with IMR-90 fibroblasts known to secrete cysteine. SASP, at 0.15 and 0.2 mM, arrested replication of fibroblast-driven Nb2-11 cells by 93 and 100%, respectively, without impeding fibroblast proliferation. Addition of 2-mercapto-ethanol (60 microM), a cystine uptake enhancer, almost completely prevented this growth arrest, indicating that SASP specifically inhibited cysteine secretion by the fibroblasts, a process based on x(c)- -mediated cystine uptake. It is proposed that the lymphoma growth-inhibitory activity of SASP in vivo involves inhibition of cysteine secretion by tumor-associated somatic cells (macrophages, dendritic cells), leading to cysteine starvation of the tumor cells and apoptosis. The difference between the lymphoma cells and fibroblasts in sensitivity to SASP treatment is consistent with the marked antitumor effect of SASP lacking significant side effects. PMID- 12544256 TI - Induction of HL-60 cell differentiation by the p38 mitogen-activated protein kinase inhibitor SB203580 is mediated through the extracellular signal-regulated kinase signaling pathway. AB - The pyridinyl imidazole p38 kinase inhibitor, SB203580, was initially used to block inflammatory cytokine synthesis. Here we report that SB203580 by itself could induce human promyeloid leukemic HL-60 cells to differentiate mainly along the granulocytic lineage, as evidenced by cellular morphological changes, and the concurrent expression of cell surface markers CD11b and CD14. This differentiation induction was time and dose dependent. After 12 h exposure to 10 microM SB203580, 12.5% of the cells became CD11b as compared to only 2.6% in untreated control cells. By 96 h, CD11b cells increased to 72.3%, and among them, 26% were CD14. Morphologically, the cells were smaller in size with lower nuclear/cytoplasmic ratio. The nucleus was indented and nucleoli markedly reduced. However, 10 microM SB203580 had little effect on HL-60 cell growth and survival during the first 72 h, but by 96 h the percentage of cells in G1 phase was markedly increased. These effects of SB203580 were not attributable to its inhibition of p38 kinase activity. Instead, the essential kinases in the extracellular signal-regulated kinase (ERK) pathway such as phospho-Raf-1, phospho-MEK1/2, phospho-ERK1/2 and phospho-p90RSK were all elevated dramatically shortly after cells were exposed to SB203580 and lasted for 24 h before declining. Pre-incubation of cells with 20 microM of PD98059 1 h before addition of SB203580 could completely block the expression of differentiation markers. Our results suggest that SB203580-induced differentiation in HL-60 cells was mediated by activation of MEK/ERK signaling. In conclusion, our data have shown that SB203580 possessed biological activities other than inhibition of p38 and these activities could make it a potential candidate as an inducing agent for cell differentiation in the therapeutic treatment of leukemia. PMID- 12544257 TI - Resistance to chemotherapeutic drugs overcome by c-Myc inhibition in a Lewis lung carcinoma murine model. AB - Chemotherapy resistance is a significant obstacle in lung cancer therapy, and has been found to frequently correlate with amplification and overexpression of the c myc oncogene. Earlier studies have shown that c-Myc inhibition alone is not always effective in cancer models. The purpose of this study was to test different dosing regimen, which included commonly used chemotherapeutic drugs in combination with c-Myc inhibition in a Lewis lung syngeneic drug-resistant murine tumor model. Inhibition of c-myc was specifically achieved by using phosphorodiamidate Morpholino oligomer (PMOs), a novel, non-toxic antisense DNA chemistry for inhibition of gene expression by an RNase H-independent mechanism. When administration of cisplatin overlapped with c-myc PMO (AVI-4126) treatment there was no additional effect on tumor growth inhibition compared to cisplatin alone. In contrast, using a dosing regimen in which cisplatin or taxol treatment preceded AVI-4126, a dramatic decrease in tumor growth rate was observed with tumor areas less then 0.5 cm2 in 60% of the animals at the end of the study. This effect was specific to c-Myc inhibition as other antisense PMOs against p21 or Rad51 showed no such effect in combination with chemotherapy. Immunoblot and HPLC based analysis of tumor lysates at the end of the study confirmed c-Myc inhibition and detection of intact AVI-4126, respectively. In conclusion, AVI 4126 potentiates the efficacy of chemotherapeutic drugs in a manner that is schedule dependent. PMID- 12544258 TI - Assessment of epidermal growth factor receptor with 99mTc-ethylenedicysteine-C225 monoclonal antibody. AB - Epidermal growth factor receptor (EGFR) plays an important role in cell division and cancer progression, as well as angiogenesis and metastasis. Since many tumor cells exhibit the EGFR on their surface, functional imaging of EGFR provides not only a non-invasive, reproducible, quantifiable alternative to biopsies, but it also greatly complements pharmacokinetic studies by correlating clinical responses with biological effects. Moreover, molecular endpoints of anti-EGFR therapy could be assessed effectively. C225 is a chimeric monoclonal antibody that targets the human extracellular EGFR and inhibits the growth of EGFR expressing tumor cells. Also, it has been demonstrated that C225, in combination with chemotherapeutic drugs or radiotherapy, is effective in eradicating well established tumors in nude mice. We have developed 99mTc-labeled C225 using ethylenedicysteine (EC) as a chelator. This study aimed at measuring uptake of 99mTc-EC-C225 in EGFR+ tumor-bearing animal models and preliminary feasibility of imaging patients with head and neck carcinomas. In vitro Western blot analysis and cytotoxicity assays were used to examine the integrity of EC-C225. Tissue distribution studies of 99mTc-EC-C225 were evaluated in tumor-bearing rodents at 0.5-4 h. In vivo biodistribution of 99mTc-EC-C225 in tumor-bearing rodents showed increased tumor-to-tissue ratios as a function of time. In vitro and biodistribution studies demonstrated the possibility of using 99mTc-EC-C225 to assess EGFR expression. SPECT images confirmed that the tumors could be visualized with 99mTc-EC-C225 from 0.5 to 4 h in tumor bearing rodents. We conclude that 99mTc-EC-C225 may be useful to assess tumor EGFR expression. This may be useful in the future for selecting patients for treatment with C225. PMID- 12544259 TI - Pharmacokinetics of doxorubicin and cisplatin used in intraoperative hyperthermic intrathoracic chemotherapy after cytoreductive surgery for malignant pleural mesothelioma and pleural thymoma. AB - Cytoreductive surgery combined with intraoperative hyperthermic intrathoracic chemotherapy (HITHOC) is studied in a phase I study in the treatment of malignant pleural mesothelioma and pleural thymoma. We studied the pharmacokinetics of doxorubicin and cisplatin used during the HITHOC procedure. Furthermore, the penetration characteristics of doxorubicin were examined. Between 1998 and 2001, 24 perfusions were performed with a solution containing doxorubicin and cisplatin for 90 min at 40-41 degrees C. The dose was first based on square meters body surface, whereas in later studies a fixed concentration of the perfusion fluid was used. Samples of blood and perfusion fluid were collected for doxorubicin and cisplatin measurements. The penetration characteristics of doxorubicin in tissue were determined by fluorescence microscopy. The mean AUC(perfusate):AUC(plasma) ratios for doxorubicin and cisplatin (ultrafiltration for plasma) were 99 and 59, respectively. During perfusion the concentration in the perfusate declined essentially according to first-order elimination kinetics for both doxorubicin and cisplatin with half-lives of 74 and 138 min, respectively. At the end of the perfusion, about 35 and 52% of the dose of doxorubicin and cisplatin, respectively, was recovered in the perfusion fluid. One patient developed a nephrotoxicity grade II. No leukopenia or hair loss was seen. Doxorubicin penetrated into the intercostal muscle specimen, albeit that there was considerable variation in distribution throughout the specimen. We conclude that HITHOC with doxorubicin and cisplatin is relatively a safe procedure with the advantage of high intrathoracic cytostatic drug concentrations, while having limited systemic side effects. PMID- 12544260 TI - A phase I study of paclitaxel, estramustine phosphate and vinorelbine (Pacl-E Vin) in advanced malignancies: triple tubulin targeting. AB - Anti-tubulin couplets have activity in hormone-resistant prostate cancer. This study was designed to define the dose-limiting toxicity (DLT) and recommended phase II dose (RPTD) of the unique triplet combination of paclitaxel, estramustine phosphate (EMP) and vinorelbine (Pacl-E-Vin). Patients with advanced malignancies who had failed standard therapy, ECOG performance status (PS 0-2) and adequate organ function were included. Dose of EMP was fixed at 300 mg/m2/dose p.o. t.i.d. on days 1-3 and 8-10. Vinorelbine dose was 20 mg/m2/day i.v. on days 3 and 10. Paclitaxel was dose escalated from 40 to 50 mg/m /day i.v. on days 3 and 10. Cycles were repeated every 3 weeks. Twelve adults (median age 72) were entered on this study. Primary tumors included prostate (n=7), cervix (n=2), melanoma (n=1), colon (1) and lung with synchronous prostate cancer (n=1). Nine patients had received no prior chemotherapy, one had received a prior regimen and two had received two or more prior regimens. Of four evaluable patients at dose level 1, one patient had grade 3 neutropenia leading to the day 10 dose being withheld. Of five evaluable patients at dose level 2, there was one DLT (febrile neutropenia) and two grade 3 neutropenias leading to the day 10 dose being withheld. One patient had a lower extremity deep vein thrombosis. Other side effects were mild and reversible. Nine patients were evaluable for efficacy: three with prostate cancer had a greater than 50% prostate-specific antigen (PSA) response, and a patient with synchronous prostate and lung cancer had a greater than 50% PSA response. We conclude that the DLT of Pacl-E-Vin is neutropenia. RPTD is vinorelbine 20 mg/m2, paclitaxel 40 mg/m2, both administered on days 3 and 10, and EMP 900 mg/m2/day on days 1-3 and 8-10, q3w. Dose omission at day 10 followed by 20% dose reduction of paclitaxel and vinorelbine is recommended in the event of grade 3 neutropenia. Activity in hormone-refractory prostate cancer is promising and warrants phase II evaluation. PMID- 12544261 TI - Docetaxel with epirubicin--investigations on cardiac safety. AB - The aim was to evaluate clinical and subclinical cardiac toxicity of epirubicin docetaxel (ET) combination. Breast cancer patients were given epirubicin (75 mg/m2 for 15 min), followed 1 h later by a 1-h infusion of docetaxel (75 mg/m2) q3w as first-line treatment. Cardiac function was monitored using a 24-h ambulatory electrocardiogram (ECG), left ventricular ejection fraction (LVEF), physical examination and chest radiography. The median LVEF did not decrease during the course of the treatment: median LVEF was 64% prior to treatment and 68% after cycle 8. The 24-h ECG did not reveal any significant changes in heart rate variability. The number of extrasystoles or cardiac arrhythmia did not increase with the ET treatment. No patient experienced congestive heart failure during treatment or the mean follow-up of 34 months. We conclude that first-line ET caused no major cardiac changes during 6 months of treatment (8 cycles) or during follow-up. Twenty-four-hour ECG, combined with echocardiography to measure LVEF, was a feasible method for the close monitoring of the cardiac effects during chemotherapy. PMID- 12544262 TI - Safety and efficacy of outpatient treatment with CPT-11 plus bolus folinic acid/5 fluorouracil as first-line chemotherapy for metastatic colorectal cancer. AB - The combination of irinotecan (CPT-11), bolus 5-fluorouracil (5-FU) and folinic acid (FA) (Saltz regimen) has recently been questioned as first-line chemotherapy for metastatic colorectal cancer after high early death rates due to gastrointestinal and thromboembolic events were reported in two US trials. Therefore, we carefully evaluated the safety and efficacy of this regimen, with high value placed on the management of delayed diarrhea. Forty-six patients with metastatic colorectal cancer received this first-line treatment in nine German outpatient clinics. Dose reductions were mandatory from the first cycle in case of toxicity grade >2. Chemotherapy was administered only to diarrhea-free patients. During a total of 175 cycles administered treatments were delayed for 1 week in 11.6% and given at a reduced dose in 14.5%. All and 40 patients were evaluable for toxicity and response, respectively. Grade 3/4 toxicities included diarrhea (n=10), leukopenia (n=9), neutropenia (n=3) and anemia (n=4). One non fatal pulmonary embolism occurred. Four complete responses (CR) and 10 partial responses were seen, for an overall response rate of 35%. In addition, 16 patients (40%) had stable disease. Resectability of liver metastases was achieved in three patients, including one pathologically confirmed CR. Median progression free and overall survival were 5 and 13 months, respectively. We conclude that outpatient treatment with the Saltz regimen was well tolerated. Severe gastrointestinal toxicity and thromboembolic events were rarely observed and never fatal. As down-staging was possible, combinations of CPT-11 and FA/5-FU should be further investigated in neoadjuvant protocols. PMID- 12544263 TI - Palliative chemotherapy with gemcitabine and weekly high-dose 5-fluorouracil as 24-h infusion in metastatic biliary tract and gall bladder adenocarcinomas. AB - At present, systemic treatment is not generally recommended for advanced biliary tract and gall bladder carcinomas. In particular cases, however, it may be justified to consider systemic chemotherapy treatment. In four cases we investigated the efficacy of palliative systemic treatment in metastatic biliary tract and gall bladder adenocarcinomas. Similar to the proceedings in a phase II study for metastatic pancreas adenocarcinomas, four patients with advanced biliary tract and gall bladder adenocarcinomas received a combination treatment of gemcitabine (GEM) and weekly high-dose 5-fluorouracil (5-FU) as a 24-h infusion. Altogether, the four patients received 96 chemotherapy applications. The palliative chemotherapy was tolerated well. In one patient, leukocytopenia (toxicity grade III) and thrombocytopenia (toxicity grade III) occurred. In three patients, the palliative systemic treatment led to stable disease, partly with a significant decrease of the CA 19-9 tumor marker, and in one patient to partial remission (PR). The survival times in these four patients were 6, 10, 17 and 26 months. Even in the case of PR, a curative hemihepatectomy right could be achieved after 'downsizing'. We conclude that in the four case studies, the applied palliative combination treatment based on GEM and 5-FU proved to be effective. However, future multicenter studies will be necessary to determine the significance of palliative chemotherapy in biliary tract and gall bladder carcinomas. PMID- 12544264 TI - Hematology and oncology. PMID- 12544265 TI - Endocrine late effects of cancer treatment. AB - Endocrinopathies are significant consequences of childhood cancers and their treatments. The risk of developing these adverse events is related to the underlying disease and its treatment with cytotoxic drugs and radiation therapy. This article will explore the current literature about endocrine late effects of hypothalamic-pituitary, thyroid, and gonadal dysfunction, as well as osteoporosis and obesity. PMID- 12544266 TI - The management of Hodgkin disease in the young child. AB - Although childhood Hodgkin disease is sensitive to the treatment regimens devised for Hodgkin disease in adults, long-term toxicity is enhanced in the developing individual. As a result, there have been dual goals in the design of clinical trials for pediatric Hodgkin disease: 1) to reduce long-term organ injury; and 2) to increase efficacy. Radiation dose and field has been reduced by enhanced reliance on chemotherapy, thus limiting the risks of hypoplasia, hypothyroidism, secondary cancers, and valvular and atherosclerotic heart disease. Multiagent, chemotherapeutic regimens for children have been developed to avoid the risks of sterility, leukemia, and cardiopulmonary toxicity. Newer approaches advocate for early dose intensity to limit cumulative therapy using response-based paradigms. Targeting molecular mechanisms specific for the Reed-Sternberg cell may allow for less toxic and more efficacious treatments in the future. PMID- 12544267 TI - Thrombocytopenia in the newborn. AB - Thrombocytopenia remains a common problem in sick newborns. A quarter of all neonates admitted to neonatal intensive care units develop thrombocytopenia, and in 20% of episodes the thrombocytopenia is severe (platelets <50 x 10(9)/L). Practical and clinically relevant classifications of neonatal thrombocytopenia have now been developed which, by highlighting the principal conditions precipitating severe thrombocytopenia (eg, sepsis, necrotizing enterocolitis, perinatal asphyxia, and the immune thrombocytopenias), aid the practicing neonatologist. Recent reviews demonstrate that many neonates with severe thrombocytopenia receive repeated platelet transfusions, although evidence of their clinical benefit is lacking, and there exists a significant variation in platelet transfusion practice between centers. These facts support the need for the development of evidence-based protocols for platelet transfusion in the newborn and stimulate continued interest in the potential of hemopoietic growth factors (, thrombopoietin and interleukin-11) to prevent or treat neonatal thrombocytopenia. PMID- 12544268 TI - Stem cell transplantation for hemoglobinopathies. AB - Hereditary anemias caused by beta-thalassemia and sickle cell disease are the most common genetic diseases worldwide. Supportive therapies such as chronic lifelong transfusions, iron chelation for thalassemia, and transfusions or hydroxyurea for sickle cell anemia have significantly ameliorated clinical manifestations of these diseases but cannot eliminate disease and treatment related complications that result in end-organ damage. Allogeneic hematopoietic stem cell transplantation is the only cure for patients with hemoglobinopathies. Results of transplants have steadily improved over the last few decades due to effective control of transplant-related complications and development of new preparative regimens. Our understandings of mixed chimerism in patients with hemoglobinopathies provide a rationale for the use of less intensive conditioning regimens and gene therapy in these disorders. Although the role of stem cell transplantation for thalassemia major is well defined, few transplants have been carried out in sickle cell disease, and, in light of recent advances, the role of stem cell transplantation in this disease should be revised. This review summarizes the current status of stem cell transplantation for hemoglobinopathies. PMID- 12544269 TI - Stem cell plasticity: a new image of the bone marrow stem cell. AB - The central tenet of stem cell biology is that within tissues there reside stem cells with the capacity for both self-renewal and terminal differentiation to the multiple lineages of that tissue. Over the last few years, numerous studies have challenged this paradigm by showing that tissue stem cells can differentiate to unexpected cell lineages, suggesting an enormous plasticity of differentiation. The hematopoietic stem cell, which resides within bone marrow and gives rise to all blood cells, has been the focal point of these efforts. However, recent studies have disputed the notion of hematopoietic stem cell plasticity. In truth, stem cell plasticity, strictly defined, has yet to be rigorously proven. Both animal models to carefully address outstanding issues and pilot clinical trials to explore the therapeutic potential will be key elements to advance science for the benefit of patients. PMID- 12544270 TI - Osteochondritis dissecans of the knee in children. AB - Osteochondritis dissecans is a term used to describe the separation of an articular cartilage subchondral bone segment from the remaining articular surface. Juvenile osteochondritis dissecans describes an osteochondritis dissecans lesion found in skeletally immature children with a maximum incidence occurring between the ages of 10 and 20. It is found more frequently in children who are active athletically and involved in organized sports and is twice as common in males as in females. Although the etiology of these lesions is unclear, it is believed that repetitive microtrauma may interrupt the already tenuous epiphyseal blood supply in the growing child and contribute to the development of osteochondritis dissecans lesions. Treatment is dependent upon age at presentation, fragment size, fragment location, and fragment stability. Stable lesions in skeletally immature patients are generally amenable to conservative management. Failed conservative management or unstable lesions will more likely require surgical intervention. Lesions in skeletally mature patients have a more unpredictable course and may require surgery. This review article discusses the anatomy, etiology, evaluation, classification, treatment, and expected outcome of osteochondritis dissecans lesions. PMID- 12544271 TI - Anterior cruciate ligament injury in children: update of current treatment options. AB - As our society's interest in competitive athletics has grown, so has the participation of our youth. Unfortunately, along with this increase in participation has come a predictable increase in rate of injury. More specifically, anterior cruciate ligament injury in the skeletally immature individual is being recognized with increasing frequency and currently poses an unsolved clinical problem. Conservative management of midsubstance anterior cruciate ligament tears in the skeletally immature population has been shown to have an unfavorable prognosis related to functional knee instability, subsequent meniscal tears, and the development of early degenerative arthritis. Despite poor outcomes following conservative treatment, many orthopaedic surgeons have been reluctant to perform anterior cruciate ligament reconstructions in skeletally immature patients due to the potential for physeal injury and resultant growth disturbance. Although there is growing evidence in the literature suggesting that anterior cruciate ligament reconstruction in the adolescent population may be safely performed using anatomic, transphyseal techniques, there are insufficient data to provide concrete guidelines in treatment of anterior cruciate ligament injuries in the prepubescent population. Management of these injuries, therefore, must be based on the physiologic and skeletal maturity of the child. Anterior cruciate ligament reconstruction in the skeletally immature individual still poses a clinical problem with the safest and most effective techniques still evolving. PMID- 12544272 TI - Ewing sarcoma: current concepts in diagnosis and treatment. PMID- 12544273 TI - Benign bone tumors in children. AB - When children present for evaluation of bone tumor, great anxiety often occurs among the family. Usually the diagnosis of a benign bone tumor can be made easily with plain radiographs without biopsy. Most have a good outcome. However, occasionally, serious complications can develop. Moreover, the physician must be aware of the possibility of the malignant tumors. To avoid errors in the evaluation and treatment of this heterogeneous group of tumors, the clinician should have a good knowledge base of common bone neoplasms and correlate the clinical, radiographic, and pathologic findings. In this article, history, diagnosis, evaluation, treatment, and management of common types of benign tumors of bone commonly seen in children and adolescents are reviewed in the light of current literature. PMID- 12544274 TI - Elbow injuries in children. PMID- 12544275 TI - New recommendations for influenza vaccination for children and pregnant women. AB - This year the Advisory Committee on Immunization Practices of the United States Centers for Disease Control and Prevention has updated recommendations for use of influenza vaccine. Previously, use of influenza vaccine focused primarily on the elderly as well as younger persons with underlying conditions that place them at high risk for severe disease and complications from influenza infection. The new recommendations also emphasize the benefits of influenza vaccination for young, healthy children who are at high risk for hospitalization with influenza infection. These changes are the result of recent reports demonstrating that otherwise healthy young children aged 6 to 24 months are hospitalized for influenza and its complications at rates comparable to those for whom influenza vaccination is already recommended, including the elderly. PMID- 12544276 TI - Rapid diagnostic tests for influenza. AB - In recent years, seven rapid tests for influenza have become commercially available. These tests offer the promise of making rapid influenza diagnosis much more widely available than in the past, when rapid diagnosis could be made only by fluorescent antibody staining, a demanding technique available only in advanced laboratories. Two of the rapid tests have waived status under the Clinical Laboratory Improvements Amendments of 1988. This article describes each of the seven tests and reviews English-language publications that have evaluated the performance of these tests on specimens from children. A discussion of the implications of the tests for clinical decision-making is included. PMID- 12544277 TI - Human herpesvirus 8 infection. AB - Human herpesvirus 8, also known as Kaposi sarcoma-associated herpesvirus, is etiologically associated with Kaposi sarcoma and other rare malignancies. Human herpesvirus 8 infection is common in certain areas of Africa and Italy, but occurs in only 0% to 15% of adult populations in North America and Europe. Reports of human herpesvirus 8 prevalence of 3% to over 50% among children in Central Africa, Brazil, and South Texas suggest that horizontal transmission of human herpesvirus 8 occurs among children. Primary human herpesvirus 8 infection in immunocompetent children is associated with a fever and maculopapular rash. PMID- 12544278 TI - Recombinant human activated protein C for the treatment of severe sepsis: is there a role in pediatrics? AB - Sepsis with organ failure (severe sepsis) remains an important cause of morbidity and mortality among children. The clinical pathophysiology of severe sepsis reflects a coordinated activation of the innate immune response, including elaboration of proinflammatory cytokines and the induction of the extrinsic pathway of coagulation (sepsis-induced coagulopathy). These proinflammatory and procoagulant pathways are linked, and are similarly coregulated by a number of proteins and factors, including protein C. However, at least 80% of children and adults with severe sepsis develop acquired deficiency of protein C because of factor consumption. This deficiency is associated with poor outcomes, including multiple organ failure and mortality. Recently, recombinant activated protein C was shown to reduce the mortality of adults with severe sepsis, and is now approved for such use in the United States and Europe. The rationale for pediatric applications of protein C and ongoing clinical trials in children are reviewed. PMID- 12544279 TI - Candida infections in the neonate. AB - Candida infections have become an increasingly frequent problem in neonatal intensive care units, particularly among extremely low birth weight infants. Transmission occurs both vertically and horizontally, with Candida albicans and C. parapsilosis as the predominant species. Multiple risk factors have been identified with prior antibiotic exposure, presence of a central line, endotracheal intubation, and prior fungal colonization reported most frequently. The primary site of infection can involve the bloodstream, meninges, or urinary tract, but disease is frequently disseminated to multiple organ systems. Amphotericin is the most commonly used antifungal agent, although fluconazole is being used more frequently. The potential role of antifungal prophylaxis is not yet clearly defined, but has been the topic of recent investigative efforts. The crude mortality rate among neonates with systemic candidiasis remains approximately 30%. PMID- 12544280 TI - Stachybotrys. AB - Stachybotrys is a toxin-producing fungus that grows indoors when both water and cellulose are available. Epidemiologic evidence has demonstrated an association between acute pulmonary hemorrhage in infants and exposure to Stachybotrys and other fungi in water-damaged home environments. In recent years, advances in understanding of this association have occurred in six major areas: animal models, biologic mechanism of lung injury, dose-response relationship, isolation from diseased patients, detection methods, and intervention. The association demonstrates strength, consistency, coherence, and specificity. While additional data are gathered, a preventive approach to reducing the exposure of infants with pulmonary hemorrhage is suggested. PMID- 12544281 TI - Ring-a-ring-a-roses: bioterrorism and its peculiar relevance to pediatrics. PMID- 12544282 TI - Fever without apparent source on clinical examination. AB - This section focuses on issues in infectious disease that are commonly encountered in pediatric office practices. Dr. McCarthy discusses recent literature regarding the evaluation and management of acute fevers without apparent source on clinical examination in infants and children and the evaluation of children with prolonged fevers of unknown origin. Drs. Klig and Chen (pp 121-126) review recent literature about lower respiratory infection in children. This section focuses on febrile children in whom a source of fever is not readily apparent on clinical examination. This issue is discussed in several contexts: recent developments concerning the epidemiology, pathophysiology, diagnostic approach, and therapy of febrile illnesses; children from 3 to 36 months of age with fever; infants younger than 90 days of age with fever; and children of any age with prolonged fever, usually lasting more than 7 to 10 days, for whom a diagnosis has not been established. PMID- 12544283 TI - Lower respiratory infections in children. AB - Lower respiratory infections (LRIs) continue to threaten the health of children worldwide and are exacerbated by global environmental problems such as air pollution [1]. In the developing world where nutrition remains poor and access to healthcare is scarce, LRIs are the most common cause of illness and death in children. Outcomes of LRI illness are far better in developed countries, but the overall morbidity of LRI is still high and may exceed that of other age groups [2]. The environmental "health" of our planet remains an issue closely linked to the health of children--particularly in terms of LRI disease. It is also clear that further progress in preventing, diagnosing, and treating LRI disease will have a significant impact on the health of children. The recent literature details both progress and clinical dilemmas of LRI-related illnesses in children. Lower respiratory infection disease is considered to include the following entities: acute lower respiratory infections, pneumonia, atypical pneumonia, bronchitis, and bronchiolitis. This update will review current reports on the evaluation of children with LRIs, bacterial and viral LRIs, and sequelae of LRIs. PMID- 12544284 TI - Case report: nonalcoholic steatohepatitis in a teenage girl with type 2 diabetes. PMID- 12544285 TI - Surgical techniques for treating a renal neoplasm invading the inferior vena cava. AB - PURPOSE: Historically inferior vena caval thrombus associated with renal cell carcinoma was a deterrent to surgery. During the last 3 decades there has been steady improvement in surgical techniques and perioperative care, which has dramatically improved the ability to resect safely these tumors. We acknowledge these improvements in chronological order. MATERIALS AND METHODS: A comprehensive literature review of the different techniques used for resecting renal cell carcinoma with inferior vena caval involvement was performed using MEDLINE. Data focused on surgical techniques, including various incisions, exposures, adjuncts to surgery and outcomes. RESULTS: Tumor thrombus associated with renal cell carcinoma is no longer considered to have a detrimental impact on survival. Patients who are acceptable surgical candidates have survival rates as high as 68%. Although there is a great deal of emphasis on the importance of an aggressive surgical approach, a uniform operative strategy based on the level of the tumor thrombus has not been established. Surgical techniques derived from liver transplant surgery and cardiac arrest with cardiopulmonary bypass have drastically decreased operative complications associated with extensive involvement of the inferior vena cava with tumor thrombus. CONCLUSIONS: The only curative approach to renal cell carcinoma is surgery. An aggressive approach is warranted when tumor involves the renal vein and inferior vena cava. Surgical strategy depends on the level of the inferior vena caval thrombus. Patients with extension of the thrombus above the diaphragm are a greater technical challenge. Hypothermic circulatory arrest should be considered when treating vena caval atrial tumor thrombus. Surgeons familiar with liver mobilization can greatly facilitate the exposure needed for safely operating in these cases. PMID- 12544286 TI - Prostate specific antigen and human glandular kallikrein 2 in early detection of prostate cancer. AB - PURPOSE: Several tumor markers have recently been applied for prostate cancer screening. We analyze the effectiveness of prostate specific antigen (PSA), age specific PSA, PSA velocity, volume adjusted PSA densities, change in PSA level following antibacterial therapy, free-to-total PSA ratio, alpha1-antichymotrypsin bound PSA, alpha2-macroglobulin bound PSA, alpha1-protease inhibitor bound PSA and human glandular kallikrein 2 in detecting prostate cancer. MATERIALS AND METHODS: We conducted a review of the literature between September 2000 and February 2001. A total of 7,250 abstracts and articles published during the previous 12 years were retrieved from MEDLINE using the key words PSA and human glandular kallikrein 2. Of these reports 135 are included in this review. RESULTS: We analyzed and systematized data from studies regarding the effectiveness of PSA and human glandular kallikrein 2 and their derivatives in the detection of prostate cancer. CONCLUSIONS: Improvement in the specificity and sensitivity of PSA is imperative. Free-to-total PSA ratio, transition zone PSA density and change in PSA level increase the specificity of PSA to some extent. Protocols investigating the effectiveness of different combinations of these 3 measurements seem necessary for improving the effectiveness of prostate cancer screening among men within the diagnostic "gray zone." PSA velocity, age adjusted PSA levels and PSA density might be used in limited cases. alpha1 Antichymotrypsin, alpha2-macroglobulin and alpha1-protease inhibitor bound PSA, and human glandular kallikrein 2 are promising experimental methods. PMID- 12544287 TI - Hugh Hampton Young: father of American urology. PMID- 12544288 TI - Prevention of stone formation and bone loss in absorptive hypercalciuria by combined dietary and pharmacological interventions. AB - PURPOSE: We determined whether dietary restriction of calcium and oxalate, combined with thiazide and potassium citrate treatment, would prevent stone formation and avert bone loss in 18 men and 10 women with type I absorptive hypercalciuria. MATERIALS AND METHODS: Patients were treated with thiazide (20) or indapamide (8) and potassium citrate (average dose 35 mEq. daily) for 1 to 11 years (mean 3.7) while maintained on low calcium oxalate diet. Serum and urinary chemistry studies and bone mineral density were measured at baseline and at the end of treatment. New stones formed were quantitated during 3 years before and during treatment. RESULTS: During treatment urinary calcium significantly decreased (346 +/- 85 to 248 +/- 79 mg. daily, p <0.001) but urinary oxalate did not change. Urinary pH and citrate significantly increased, and urinary saturation of calcium oxalate significantly decreased by 46%. Stone formation rate decreased significantly from 2.94 to 0.05 per year (p <0.001). L2-L4 bone mineral density increased significantly by 5.7% compared to normal peak value, and by 7.1% compared with normal age and gender matched value. Femoral neck bone mineral density also increased significantly. CONCLUSIONS: Dietary restriction of calcium and oxalate, combined with thiazide and potassium citrate, satisfactorily controlled hypercalciuria, prevented the secondary increase in urinary oxalate, reduced urinary saturation of calcium oxalate, virtually eliminated recurrent stone formation, and increased bone density of the spine and femoral neck. Thus, this dietary pharmacological program controlled stone formation as well as bone loss that often accompany type 1 absorptive hypercalciuria. PMID- 12544289 TI - Effect of dietary calcium on stone forming propensity. AB - PURPOSE: Epidemiological studies have reported that high calcium diet protects against kidney stone formation in normal subjects. This metabolic study was designed to elucidate the physiological and physicochemical effects conferring this apparent protection. MATERIALS AND METHODS: A total of 21 normal volunteers underwent 2 phases of study in a crossover, randomized design, wherein they consumed constant metabolic diets that matched the estimated highest and lowest quintiles of calcium intake from published epidemiological studies. RESULTS: Urinary calcium was significantly greater on the high calcium diet (148 +/- 55 versus 118 +/- 43 mg. daily, p <0.01, p <0.01) but urinary oxalate did not differ between diets. There was no difference in relative saturation ratio of calcium oxalate between the 2 diets. The high calcium diet significantly increased saturation of brushite and decreased that of uric acid. Due to the other differences between the diets (more fluid, potassium, magnesium and phosphate in the high calcium diet), the high calcium diet also increased 24-hour urinary volume, potassium, phosphorus, pH and citrate. After adjustment of these confounding variables, the high calcium diet significantly increased relative saturation ratio of calcium oxalate by 24%. CONCLUSIONS: High calcium diet from published epidemiological studies does not alter the propensity for calcium oxalate crystallization in normal subjects despite increased urinary calcium and unaltered urinary oxalate because of the greater amounts of ingested fluid, potassium and phosphate. However, high calcium intake alone, without concomitant changes in the diet, poses a modest risk for calcium stone formation. PMID- 12544290 TI - The etiology of urolithiasis in HIV infected patients. AB - PURPOSE: It is commonly thought that urinary lithiasis in HIV infected patients on protease inhibitor therapy is composed primarily of the protease inhibitor itself. Since many HIV infected patients on protease inhibitors presenting to our institution had nonprotease inhibitor stones, we investigated potential underlying metabolic abnormalities that may account for the lithogenesis. MATERIALS AND METHODS: We retrospectively reviewed all HIV infected patients on protease inhibitors with renal colic and evidence of nephrolithiasis who presented to our institution between June 1996 and January 2001. Patients were evaluated for stone composition and metabolic abnormalities of blood and urine when possible. RESULTS: A total of 24 patients were identified, and all were or had been on protease inhibitors (indinavir 14, ritonavir 3, nelfnavir 2, unspecified 5). Of the 14 patients on indinavir only 4 (28.6%) had indinavir containing stones. The remaining stones in this group and in those not on indinavir contained various amounts of calcium oxalate monohydrate and dihydrate, ammonium acid urate and uric acid. Of 10 patients who underwent 24-hour urine collection for metabolic evaluation 8 (80%) had abnormalities, including hypocitraturia in 5, hyperoxaluria in 4, hypomagnesuria in 4, hypercalciuria in 3, increased supersaturation of calcium oxalate in 3 and hyperuricosuria in 2. Abnormalities in the levels of urinary phosphate and sodium were also observed. CONCLUSIONS: HIV infected patients form many types of stones, which probably are attributable to underlying metabolic abnormalities rather than the use of protease inhibitors. A complete metabolic evaluation is warranted in these patients, as a means of guiding treatment to prevent future stone episodes, while avoiding the need to alter antiretroviral regimens. PMID- 12544291 TI - Editorial: Something new about stones. PMID- 12544292 TI - Minimally invasive retroperitoneoscopic ureterolithotomy. AB - PURPOSE: We assessed the efficacy of modified technique of retroperitoneal ureterolithotomy for managing ureteral stones. MATERIALS AND METHODS: Between December 1999 and March 2002, 31 patients underwent retroperitoneal ureterolithotomy. In most cases only 3 ports (a 10 mm., 2 5 mm. and in a few 2, 3 mm.) were used without any ureteral stent or catheter. The stone was removed from the primary port site, while visualizing retrieval through the 3/5 mm. port using fine laparoscope. Ureterotomy closure was performed by intracorporeal interrupted sutures of 4-zero polyglactin. RESULTS: The 20 males and 11 females had a mean age of 38.5 years. Mean operative time was 67 minutes and mean hospital stay was 2.4 days. The mean analgesic requirement was 42.2 mg. meperidine. Mean followup was 8 months. There were no significant postoperative complications except persistent urine leakage in 2 patients 48 hours after surgery. An internal stent was placed and leakage subsided without any consequences. CONCLUSIONS: The modified, minimally invasive technique of retroperitoneal ureterolithotomy is highly effective and efficient without an associated increase in the complication rate. PMID- 12544293 TI - Laparoscopic partial nephrectomy for renal tumor: single center experience comparing clamping and no clamping techniques of the renal vasculature. AB - PURPOSE: We performed a nonrandomized retrospective comparison of 2 techniques for laparoscopic partial nephrectomy, that is without and with clamping the renal vessels. MATERIALS AND METHODS: Between December 1997 and February 2002, 28 consecutive patients underwent transperitoneal laparoscopic partial nephrectomy for renal tumor. In group 1 (12 patients) partial nephrectomy was performed with ultrasonic shears and bipolar cautery without clamping the renal vessels, while in group 2 (16 patients) the renal pedicle was clamped before tumor excision. In group 2 patients intracorporeal kidney cooling was achieved by a ureteral catheter connected to 4C solution. Intracorporeal freehand suturing techniques were used to close the collecting system when opened and approximate the renal parenchyma. RESULTS: All procedures were successfully completed laparoscopically. Mean renal ischemia time +/- SD was 27.3 +/- 7 minutes (range 15 to 47) in group 2 patients. Mean laparoscopic operating time was 179.1 +/- 86 minutes (range 90 to 390) in group 1 compared with 121.5 +/- 37 minutes (range 60 to 210) in group 2 (p = 0.004). Mean intraoperative blood loss was significantly higher in group 1 than in group 2 (708.3 +/- 569 versus 270.3 +/- 281 ml., p = 0.014). Three patients in group 1 and 2 in group 2 required blood transfusions. Immediately postoperatively mean creatinine was 1.26 +/- 0.36 and 1.45 +/- 0.61 mg./dl. in groups 1 and 2, respectively (p = 0.075). Surgical margins were negative in all specimens. Pathological examination revealed renal cell cancer in 18 cases (stages pT1 in 17 and pT3a in 1), oncocytoma in 4, angiomyolipoma in 5 and renal adenoma in 1. CONCLUSIONS: Laparoscopic partial nephrectomy represents a feasible option for patients with small renal masses. Clamping the renal vessels during tumor resection and suturing the kidney mimics the open technique and seems to be associated with less blood loss and shorter laparoscopic operative time. PMID- 12544294 TI - Laparoscopic radical nephrectomy for cancer with level I renal vein involvement. AB - PURPOSE: Venous involvement develops in 5% to 10% of patients with renal cell carcinoma and is generally considered a relative contraindication to laparoscopic radical nephrectomy. To our knowledge we report the initial clinical series of laparoscopic radical nephrectomy for renal cell carcinoma associated with level I renal vein thrombus. MATERIALS AND METHODS: At our 2 institutions 8 patients each underwent laparoscopic radical nephrectomy for level I microscopic renal vein thrombus (group 1) and level I gross thrombus (group 2). In all 8 group 2 patients the level I thrombus was preoperatively diagnosed by computerized tomography. Mean renal tumor size in groups 1 and 2 was 7.8 and 12.4 cm., respectively. After controlling the renal artery the renal vein was secured by firing an endoscopic gastrointestinal anastomosis stapler on its collapsed, uninvolved proximal part adjacent to the vena cava. Intraoperative, postoperative and pathological parameters were assessed in the 2 groups. RESULTS: In group 1 laparoscopic radical nephrectomy was technically successful in all 8 patients. Mean operative time was 3.1 hours, mean estimated blood loss was 382 cc and mean hospital stay was 1.9 days. In 1 patient each a soft tissue and a vascular margin was positive for cancer. At a mean follow up of 19.5 months (range 2 to 36) metastatic disease occurred in 3 cases (38%). In group 2 laparoscopic radical nephrectomy was technically successful in 7 cases with open conversion in 1. Mean operative time was 3.3 hours, mean estimated blood loss was 354 cc and mean hospital stay was 2.3 days. Surgical soft tissue and the renal vein vascular margin of the transected vein were negative for cancer in all 8 cases. At a mean followup of 9.4 months (range 5 to 16) pulmonary metastasis developed in 1 patient (13%). CONCLUSIONS: Although it is an advanced procedure, laparoscopic radical nephrectomy in patients with level I renal vein thrombus is feasible, safe and follows established oncological principles. PMID- 12544295 TI - Prognostic implications of p53 gene mutations in bladder tumors. AB - PURPOSE: Alterations in the p53 gene related to neoplastic progression were studied in tumor tissue samples from patients with transitional cell carcinoma and correlated with classic staging parameters. On this basis, biological characterization of the tumor was performed to establish subgroups of patients at high risk and those with a more favorable prognosis. MATERIALS AND METHODS: This observational, analytical and cross-sectional study included 115 patients divided into 4 homogeneous groups of 1-control, 2-primary superficial transitional cell carcinoma, 3-recurrent superficial transitional cell carcinoma, and 4 infiltrative transitional cell carcinoma. DNA was obtained from tumor tissue samples and polymerase chain reaction-single strand conformational polymorphism analysis was performed on exons 5 to 9 of the p53 gene. Samples showing mutations were submitted to automatic sequencing. Statistics included bivariate analysis and logistic regression. RESULTS: Of the tumors the 63.8% were superficial and 37.2% were infiltrative transitional cell carcinoma. Of the infiltrative tumors 23.5% (8 of 34) resulted from recurrent transitional cell carcinoma. Mutations were found in samples from 46.8% of patients, all with bladder tumors. There was a trend toward increasing appearance of mutations as the size of the tumor, number of tumor implants, degree of dedifferentiation and stage of local infiltration increased. The presence of mutations in p53 was 2.5 times greater in infiltrative tumors than in low stage and 4.3 times greater in moderate to high grade than in low grade tumors. All mutations found were point mutations and 79.25% provoked severe alterations in protein structure. CONCLUSIONS: Mutations in the p53 gene are mainly point mutations that aggregate in hot spots, and provoke genetic instability and substantial changes that alter p53 function, implying a trend to tumor progression and dissemination (with a greater proportion of mutations in high stage high grade tumors). Since a large percentage of bladder tumors are under staged, analysis of p53 gene mutations could be useful as a factor for prognosis and therapeutic decisions. PMID- 12544296 TI - Multi-institutional long-term experience with conservative surgery for invasive penile carcinoma. AB - PURPOSE: Invasive squamous cell carcinoma of the penis occurs on the glans, prepuce, glans and prepuce, coronal sulcus and shaft. Penile squamous cell carcinoma subsequently invades local structures, corpora cavernosa and the urethra, and metastasizes to the inguinal lymph nodes. Invasive squamous cell carcinoma of the penis usually requires total or partial penectomy. We studied the effect of primary tumor resections tailored to the anatomical extent of the cancer with preservation of uninvolved structures in select patients with invasive penile squamous cell carcinoma. MATERIALS AND METHODS: A total of 30 patients between 39 and 82 years old were treated with unconventional conservative surgical excision of the primary penile lesion. More than 130 patients were excluded from the study because they were treated with partial or total penectomy, Mohs' surgery or more extensive surgery. The 30 patients underwent preoperative biopsy with careful mapping of the extent of the disease. Patient age, tumor extent and grade, operative details, outcome and length of followup were analyzed. RESULTS: Tumor size ranged from 1.5 to 8 cm. in diameter. Tumors were well differentiated in 19 patients, moderately differentiated in 5 and poorly differentiated in 6. A total of 17 patients underwent ilioinguinal lymphadenectomy, 12 of whom had pathologically positive lymph nodes. Inguinal radiation was used in 2 patients. Chemotherapy was given to 7 patients with extensive inguinal lymphadenopathy and to 2 of 5 with pathologically positive lymph nodes. Followup ranged from 12 to 360 months. A total of 21 patients had no evidence of disease at last followup. Tumor resection with no sacrifice of function was performed in 2 patients in whom 3 small recurrences developed. One patient with numerous tumors had 2 small recurrences, which were completely excised with no further recurrence. Of the 7 patients with advanced lymphadenopathy 5 and of 5 patients with pathologically positive lymph nodes at presentation 1 died of the cancer but had no local recurrence in the penis. CONCLUSIONS: In a minority of patients with anatomically suitable penile cancer conservative surgical techniques are safe and provide equal tumor control compared to conventional resections. The anatomical situation and tumor characteristics should dictate the choice of treatment for the primary penile lesion. Inguinal lymph nodes should be managed by appropriately established guidelines but should not influence the extent of primary penile lesion resection. PMID- 12544297 TI - Does retroperitoneal lymphadenectomy for testicular germ cell tumor require a different approach in the presence of horseshoe kidney? AB - PURPOSE: We report our management of stage II testicular nonseminomatous germ cell tumor in 2 patients with horseshoe kidney and discuss the technical challenges posed by this renal fusion anomaly. The embryology and clinical anatomy of horseshoe kidney are discussed with particular reference to the anomalous vascular pattern and routes of testicular lymphatic drainage in this setting. Modifications and innovations of the standard technique of retroperitoneal lymphadenectomy in the presence of horseshoe kidney are discussed in light of our experience with these patients at 2 major tertiary care cancer centers. The significance of contemporary advanced noninvasive radiological techniques, such as helical computerized tomographic angiography with digital 3 dimensional reconstruction and magnetic resonance angiography, in the surgical planning and safe performance of surgery is emphasized. MATERIALS AND METHODS: Two young male patients treated at 2 major American teaching hospitals who had coexistent stage II testicular nonseminomatous germ cell tumor and horseshoe kidney underwent salvage retroperitoneal lymph node dissection. RESULTS: There was no evidence of recurrence in these 2 patients 12 and 15 months after surgery, respectively. CONCLUSIONS: Horseshoe kidney poses special technical problems during retroperitoneal lymphadenectomy for testicular tumors due to anomalous renal and intra-abdominal vascular patterns. Helical computerized tomography angiography is useful for meticulous surgical planning and the safe performance of surgery in this setting. PMID- 12544298 TI - Soy consumption and phytoestrogens: effect on serum prostate specific antigen when blood lipids and oxidized low-density lipoprotein are reduced in hyperlipidemic men. AB - PURPOSE: Herbal remedies high in phytoestrogens have been shown to reduce serum prostate specific antigen (PSA) and have been proposed as a treatment for prostate cancer. Soy proteins used to lower serum cholesterol are rich sources of phytoestrogens. Therefore, we assessed the effect of soy consumption on serum PSA in men who had participated in cholesterol lowering studies. MATERIALS AND METHODS: For 3 to 4 weeks 46 healthy middle-aged men with a range of starting PSA values took soy (mean 44 gm. soy protein daily, 116 mg. isoflavones daily) or control foods, and a subgroup of men took a lower level of soy supplements for 3 months. PSA was measured at the start and end of each treatment. RESULTS: Soy had no significant effect on serum total or free PSA, independent of PSA starting value or isoflavone intake. The lack of effect on PSA was seen, although soy intake was sufficient to reduce low-density lipoprotein cholesterol (5.8 +/- 2.2%, p = 0.012), the estimated coronary heart disease risk (6.1 +/- 2.8% for 10 years, p = 0.032) and the serum concentration of oxidized low-density lipoprotein measured as conjugated dienes (9.5 +/- 3.4%, p = 0.008) in the 3 to 4-week study. In addition, the lack of effect of soy on PSA persisted for the 3 months of the extended study. CONCLUSIONS: At levels of soy intake which reduce low-density lipoprotein cholesterol any potential benefits of soy consumption on prostate cancer are likely to occur for reasons other than alterations in hormone activity. PMID- 12544299 TI - Association of cigarette smoking with extraprostatic prostate cancer in young men. AB - PURPOSE: A few recent studies have revealed that cumulative or recent smoking is associated with death from prostate cancer suggesting that smoking may influence progression to more advanced disease. We evaluate the association of cigarette smoking with extraprostatic and/or Gleason sum 7 or greater prostate cancer in young men. MATERIALS AND METHODS: The study included men who underwent radical prostatectomy before age 55 years for prostate cancer between 1992 and 1999. A survey soliciting cigarette smoking history and other exposures was mailed to 498 eligible men. The response rate was 73%. Cases were defined as men with Gleason sum 7 or greater, extraprostatic or Gleason sum 7 or greater/extraprostatic disease based on pathologic analysis. All remaining participants were considered noncases for each case definition. We used logistic regression modeling to estimate the odds ratio (OR) for Gleason sum 7 or greater, extraprostatic and Gleason sum 7 or greater/extraprostatic disease with cigarette smoking. RESULTS: Of the 352 respondents with a cigarette smoking history 5.4% were current smokers and 44.6% were former smokers at the time of surgery. The odds ratios of extraprostatic and Gleason sum 7 or greater/extraprostatic disease were 3.85 (95% CI 1.44-10.33) and 3.17 (95% CI = 1.13-8.85), respectively, for current smokers compared to men who never smoked. Evidence of an association of smoking with Gleason sum 7 or greater disease was limited. Risk of extraprostatic (p = 0.005) and Gleason sum 7 or greater/extraprostatic (p = 0.003) disease increased with increasing cumulative pack-years smoked. Higher cumulative smoking in the 10 years before surgery was associated with an increased risk of extraprostatic (p = 0.004) and Gleason sum 7 or greater/extraprostatic (p = 0.005) disease. CONCLUSIONS: Cigarette smoking may influence the risk of extraprostatic prostate cancer in young men. PMID- 12544300 TI - Biochemical (prostate specific antigen) recurrence probability following radical prostatectomy for clinically localized prostate cancer. AB - PURPOSE: We retrospectively reviewed the clinical followup for a large series of men with clinically localized prostate cancer who underwent radical retropubic prostatectomy to identify clinical and/or pathological indicators of biochemical (prostate specific antigen [PSA]) recurrence. We then used those indicators to develop multivariate models for determination of recurrence probability following radical retropubic prostatectomy. MATERIALS AND METHODS: From 1982 to 1999, 2,091 consecutive men underwent radical retropubic prostatectomy and pelvic lymphadenectomy for clinically localized adenocarcinoma of the prostate (clinical stage T1c or T2 disease with Gleason score 5 or greater). Actuarial analysis was performed comparing freedom from biochemical recurrence after radical retropubic prostatectomy (PSA 0.2 ng./ml. or greater.) using the Kaplan-Meier method. Event time distributions for the time to recurrence were compared using the log rank statistic or the Cox proportional hazards regression model. The first model was developed using preoperative variables only and the second model using all available variables. Observed and predicted recurrence-free survival curves for different models were compared to select a model for calculation of predicted recurrence-free probabilities and confidence intervals. RESULTS: With a median followup of 5.9 years (range 1 to 17) 360 men (17%) had biochemical recurrence. Overall actuarial 5, 10 and 15-year biochemical recurrence-free survival rates were 84%, 72% and 61%, respectively. The relative risk of biochemical recurrence following surgery decreased with time, even after adjusted for other perioperative parameters. Variables identified for the preoperative model were biopsy Gleason score, clinical TNM stage and PSA. Variables identified for the postoperative model were prostatectomy Gleason score, PSA and pathological organ confinement status. Nomograms were generated and corrected for the decreasing relative risk of biochemical recurrence over time. CONCLUSIONS: Using 3 preoperative or postoperative parameters, these nomograms can easily be used to determine the 3, 5, 7 and 10-year biochemical recurrence-free survival probabilities among men who undergo radical retropubic prostatectomy for clinically localized prostate cancer in the modern era. PMID- 12544301 TI - Imaging findings of urinary tuberculosis on excretory urography and computerized tomography. AB - PURPOSE: We performed a retrospective study to analyze findings on excretory urography (IVP) and computerized tomography (CT) in patients with urinary tuberculosis. MATERIALS AND METHODS: In a 16-year period 53 patients with a total of 47 IVP and 33 CT examinations were diagnosed with urinary tuberculosis at our hospital. IVP and CT were reviewed and compared for certain imaging findings, including moth-eaten calices, renal parenchymal masses, an amputated infundibulum, autonephrectomy, thick urinary tract walls, urinary tract calcifications, renal parenchymal cavities, hydrocalycosis, hydronephrosis or hydroureter due to stricture, extra-urinary tubercular manifestations and renal parenchymal scarring. RESULTS: The most common finding on IVP was hydrocalycosis, hydronephrosis or hydroureter due to stricture, whereas renal parenchymal scarring was the most common finding on CT. Imaging findings of renal parenchymal masses and scarring, thick urinary tract walls and extra-urinary tubercular manifestations were significantly more common on CT than on IVP. Three imaging patterns were noted on all 44 IVPs (100%) and 31 of 33 CTs (94%) with multiple imaging findings, including multiple stricture sites, a single stricture with 1 other imaging finding and autonephrectomy with another imaging finding other than stricture. CONCLUSIONS: When the 3 imaging patterns are shown on IVP and CT, tubercular cultures or biopsies are suggested to make the definite diagnosis of urinary tuberculosis. Thus, treatment can be initiated as early as possible. PMID- 12544302 TI - Urodynamic classification of patients with symptoms of overactive bladder. AB - PURPOSE: We describe a new classification of patients with overactive bladder symptoms. MATERIALS AND METHODS: The office records of 132 patients who presented for evaluation of symptoms of urinary frequency, urinary urgency and/or urge incontinence, and who underwent videourodynamics were identified. All patients completed a 24-hour voiding diary and 24-hour pad test. Data collection included age, sex, hormone status, number of voids and incontinence episodes per 24 hours, functional bladder capacity, pad weight and associated genitourinary conditions. Videourodynamics were reviewed and patients were divided into 4 clinical categories, including type 1-no evidence of involuntary detrusor contractions on videourodynamics, type 2-involuntary detrusor contractions present, and patient aware and able to abort them, type 3-contractions present, patient aware and able to contract the sphincter but not abort contractions and type 4-contractions present and patient unaware but unable to contract the sphincter or abort contractions. RESULTS: Average patient age +/- SD was 64 years +/- 13. There were an average of 13 +/- 5 voids and 3 +/- 5 incontinence episodes per 24 hours. Average functional bladder capacity was 306 +/- 146 cc and average pad weight was 94 +/- 165 gm. Associated diagnoses included benign prostatic hypertrophy in 28% of cases, sphincteric incontinence in 17%, idiopathic urge incontinence in 29% and uterovaginal or bladder prolapse in 17%. Another 11% of patients had bladder outlet obstruction, impaired detrusor contractility or neurogenic bladder conditions. Of the cases 72 (55%), 32 (25%), 23 (17%) and 5 (4%) were categorized as classes 1 to 4, respectively. ANOVA revealed no statistically significant differences in the number of voids or incontinence episodes, functional bladder capacity or pad test when individual categories were compared to each other. CONCLUSIONS: This overactive bladder classification stratifies patients according to degrees of awareness, and control of bladder and sphincter function. It may prove useful as a guide for prognosis and therapy. Patients can be stratified into clinical groups based on the presence or absence of involuntary detrusor contractions, the ability to abort contractions and the ability to contract the urinary sphincter in response to contractions. Limiting the definition of overactive bladder to apply only to patients with no proved infection or other pathological condition would have eliminated more than 75% of those in this sample with symptoms of urinary urgency, frequency and/or urge incontinence. PMID- 12544303 TI - The evolution of detrusor overactivity after watchful waiting, medical therapy and surgery in patients with bladder outlet obstruction. AB - PURPOSE: We analyzed the evolution of detrusor overactivity in patients with bladder outlet obstruction treated with either medical or surgical therapy or watchful waiting. MATERIALS AND METHODS: Of 255 patients with symptomatic benign prostatic enlargement who completed the International Prostate Symptom Score and underwent full urodynamic investigation 161 presented with bladder outlet obstruction. Of the 161 men 101 were reevaluated with a second clinical evaluation and urodynamics 1 to 5 years (mean 2) after watchful waiting in 20, medical treatment (alfuzosin 20 and finasteride 16) in 36 and surgery (transurethral incision of the prostate 13 and prostatectomy 32) in 45. For statistical analysis Wilcoxon matched paired data and Kruskal Wallis tests were used as appropriate. RESULTS: Overall detrusor overactivity was present in 53 patients (52%) at baseline and 41 (40%) at followup. Detrusor overactivity was present in 9 patients (45%) at baseline and 11 (55%) at followup in the watchful waiting group (p = 0.17); 7 (35%) at baseline and 6 (30%) at followup in the alfuzosin group (p = 0.37); 10 (62.5%) at baseline and 10 at followup in the finasteride group (p = 1); 6 (46%) at baseline and 4 (30%) at followup in the transurethral prostate incision group (p = 0.48); and 21 (68%) at baseline and 10 (31%) at followup in the prostatectomy group (p = 0.02). CONCLUSIONS: Detrusor overactivity is highly prevalent (52%) in patients with bladder outlet obstruction, and appears to persist for long periods when obstruction is left untreated or treated only with medical therapy. However, surgical treatment of bladder outlet obstruction, prostatectomy in particular, significantly reduces the incidence of detrusor overactivity and lessens the chance of its de novo appearance for up to 5 years. PMID- 12544304 TI - Structural basis of neurogenic bladder dysfunction. I. Methods of prospective ultrastructural study and overview of the findings. AB - PURPOSE: We standardize procedures for ultrastructural study of detrusor smooth muscle and intrinsic nerves in neurogenic bladder dysfunction in the human, and present an overview of the findings. MATERIALS AND METHODS: The study included 18 female and 33 male patients 7 to 96 years old. They had neurogenic bladder dysfunction with hyperreflexia for less than 1 to 43 years, resulting from upper motoneuron lesions (spinal cord injury 25, brain disorder 17) or combined upper and lower motoneuron deficit (meningomyelocele 9). Endoscopic or open bladder biopsies were processed for ultrastructural study of detrusor smooth muscle and intrinsic neural elements. Qualitative morphologic criteria of muscle cell arrangement, degeneration and cell-cell contacts, as well as those of degeneration and regeneration of intrinsic neural elements are defined. RESULTS: Five biopsies from the brain disorder group had insufficient smooth muscle and were excluded from study. The remaining 46 biopsies were evaluated by electron microscopy, and all displayed the complete dysjunction pattern of detrusor overactivity. Most displayed degeneration and regeneration of intrinsic axons but disproportionately limited muscle cell degeneration, irrespective of detrusor contractility. The brain disorder group biopsies displayed many more ultrastructurally normal axons than the meningomyelocele and spinal cord injury group biopsies (median 33% versus 8% or less). CONCLUSIONS: Upper motoneuron neurogenic bladder dysfunction in humans is associated with intrinsic neuromuscular defects in the detrusor. Ultrastructural features of these defects suggest morphologic markers that not only may distinguish neuropathic from nonneuropathic bladder dysfunction, but also may point to the anatomical level of the neurogenic deficit. PMID- 12544305 TI - Structural basis of neurogenic bladder dysfunction. II. Myogenic basis of detrusor hyperreflexia. AB - PURPOSE: We describe the ultrastructure of detrusor smooth muscle in long standing neurogenic bladder dysfunction in the human. MATERIALS AND METHODS: Detrusor biopsies were obtained from (15 female and 31 male) patients 7 to 96 years old with neurogenic bladder dysfunction for less than 1 to 43 years. Of the patients 9 had meningomyelocele, 25 spinal cord injury and 12 brain disorder. Urodynamically, all patients had detrusor hyperreflexia (neurogenic detrusor overactivity) in addition to bladder outlet obstruction in 4, impaired detrusor contractility in 19, decreased bladder compliance in 4, and detrusor-sphincter dyssynergia in 24. Ultrastructural changes in detrusor, including those associated with detrusor overactivity, impaired detrusor contractility and bladder outlet obstruction, were evaluated qualitatively and quantitatively. RESULTS: Intermediate junctions of muscle cells were absent or reduced in 45 biopsies, which instead had dominant intimate cell appositions with much narrower junctional gaps. A greater than 2 intimate cell apposition-to-intermediate junction ratio was present in 45 biopsies (98%), and intimate cell apposition linked chains of 5 muscle cells or greater in all biopsies (100%). Muscle cell degeneration was observed in 34 biopsies from 20 of 27 patients (74%) with normal contractility and 14 of 19 (74%) with impaired detrusor contractility. No particular changes were associated with functional bladder outlet obstruction due to detrusor-sphincter dyssynergia. CONCLUSIONS: The ultrastructural complete dysjunction pattern is a feature of hyperreflexia as well as nonneuropathic detrusor overactivity of various etiology. A greater than 2 intimate cell apposition-to-intermediate junction ratio had 98% sensitivity but its specificity remains to be determined. The lack of relationship between muscle cell degeneration and detrusor contractility probably reflects limitations of urodynamic measurement of contractility in patients with spinal cord injury and meningomyelocele. PMID- 12544306 TI - Structural basis of neurogenic bladder dysfunction. III. Intrinsic detrusor innervation. AB - PURPOSE: We studied the ultrastructure of intrinsic detrusor innervation in long standing neurogenic bladder dysfunction in the human. MATERIALS AND METHODS: Endoscopic or open detrusor biopsies were obtained from 15 female and 31 male patients 7 to 96 years old who had hyperreflexic neurogenic bladder dysfunction for less than 1 to 43 years. Of the patients 9 had meningomyelocele, 25 had spinal cord injury and 12 had a brain disorder. Changes in intrinsic detrusor nerves were evaluated by electron microscopy qualitatively and quantitatively according to predefined criteria. RESULTS: Axonal degeneration was observed in 44 of the 45 biopsies with discernible intrinsic nerves. Structurally normal axons were 5(1/2) or 4 times more common in brain disorder than meningomyelocele or spinal cord injury group biopsies (median 33%, 6%, 8%, respectively). Axonal regeneration, not encountered in nonneuropathic dysfunctional detrusors, was observed in restricted distribution in most biopsies (76%) and was independent of the duration of neurogenic bladder dysfunction. Axon sprouts were observed in 17 biopsies (38%), and copeptidergic axons formed 20% (median per biopsy) of discernible axon profiles in contrast to less than 1% in normal detrusor. Activated Schwann cells were observed in all but 1 biopsy. The axonal changes were not associated with the level or degree of spinal cord lesion in patients with meningomyelocele or spinal cord injury. CONCLUSIONS: Combined degeneration and regeneration is the characteristic change in intrinsic nerves of detrusor in upper motoneuron neurogenic bladder dysfunction. The observed changes offer the possibility of clinically recognizing neuropathic contribution to a dysfunctional detrusor, as well as the potential to distinguish its spinal versus supraspinal etiology. PMID- 12544307 TI - Obstructive uropathy induced bladder dysfunction can be reversible: bladder compliance measures before and after treatment. AB - PURPOSE: We demonstrated that abnormal bladder compliance in the setting of obstructive uropathy can be improved by relief of bladder outlet obstruction. MATERIALS AND METHODS: A cohort of 9 men with nonneurogenic lower urinary tract symptoms and videourodynamics proven bladder outlet obstruction were identified prospectively from a university urology practice. Study exclusion criteria ensured absence of active urinary infection, hematuria and neurourological pathology. Testing specifically focused on assessment of the bladder compliance curve, and a compliance value was calculated (ml./cm. H2O). Treatment intervention consisted of transurethral incisions or resection of the prostate in 8 cases and transurethral balloon dilation of a urethral stricture in 1. Followup videourodynamics testing was performed 1 month after treatment to confirm relief of outlet obstruction and reassess bladder compliance. RESULTS: Mean patient age was 75.2 +/- 6.16 years. Pretreatment mean bladder compliance +/- SE was 3.06 +/- 0.45 ml./cm. H2O. At 1 month after treatment videourodynamics testing confirmed relief of obstruction in the cohort. Posttreatment mean bladder compliance +/- SE was 13.53 +/- 0.45 ml./cm. H2O. Nonparametric paired t test analysis determined that the difference between pretreatment and posttreatment bladder compliance was statistically significant at p = 0.0117. CONCLUSIONS: This pilot study suggests that relief of obstructive uropathy even in elderly patients with long-standing lower urinary tract symptoms, can significantly improve bladder compliance. PMID- 12544308 TI - Urethral atrophy after artificial urinary sphincter placement: is cuff downsizing effective? AB - PURPOSE: We reviewed the outcome of cuff downsizing with an artificial urinary sphincter for treating recurrent incontinence due to urethral atrophy. MATERIALS AND METHODS: We analyzed the records of 17 patients in a 7-year period in whom clinical, radiological and urodynamic evidence of urethral atrophy was treated with cuff downsizing. Cuff downsizing was accomplished by removing the existing cuff and replacing it with a 4 cm. cuff within the established false capsule. Incontinence and satisfaction parameters before and after the procedure were assessed by a validated questionnaire. RESULTS: Mean patient age was 70 years (range 62 to 79). Average time to urethral atrophy was 31 months (range 5 to 96) after primary sphincter implantation. Mean followup after downsizing was 22 months (range 1 to 64). Cuff downsizing caused a mean decrease of 3.9 to 0.5 pads daily. The number of severe leakage episodes decreased from a mean of 5.4 to 2.1 The mean SEAPI (stress leakage, emptying, anatomy, protection, inhibition) score decreased from 8.2 to 2.4. Patient satisfaction increased from 15% to 80% after cuff downsizing. In 1 patient an infected cuff required complete removal of the device. CONCLUSIONS: Patient satisfaction and continence parameters improved after cuff downsizing. We believe that this technique is a simple and effective method of restoring continence after urethral atrophy. PMID- 12544309 TI - Surgical intervention for complications of tension-free vaginal tape procedure. AB - PURPOSE: The tension-free vaginal tape procedure has become a state of the art operation for female stress urinary incontinence. Cases of complications requiring surgical revision are reported to be rare. We report on 6 patients with complications necessitating surgery. MATERIALS AND METHODS: Six patients who previously underwent the tension-free vaginal tape procedure required surgical management of complications, including intravesical polypropylene mesh tape with incrustation and chronic urinary tract infection in 2, vaginal mucosal mesh erosion of the vaginal incision in 1 and permanent urinary retention in 3. RESULTS: The intravesical tapes were resected via a suprapubic approach. In the case of disturbed wound healing the periurethral part of the tape was resected transvaginally. A patient in urinary retention underwent resection of the periurethral sling, while in the other 2 the tapes were transected transvaginally. Two patients in whom incontinence recurred were successfully treated with a repeat tension-free vaginal tape procedure during followup. CONCLUSIONS: Complications of the tension-free vaginal tape procedure that require surgical intervention are rare. The surgeon must be aware that this operation may lead to an additional surgical procedure, significantly increasing morbidity. PMID- 12544310 TI - Heterogeneity of 5 alpha-reductase gene expression in benign prostatic hyperplasia. AB - PURPOSE: The search for molecular markers of benign prostatic hyperplasia in general is based on an analysis of a limited number of biopsy samples. Little is known about the homogeneity of the expression of key genes in different zones of the prostate. We studied the intraprostatic (that is within the same gland) and inter-prostatic (that is between glands) variability of 5 alpha-reductase 2 (5aR2) gene expression. MATERIALS AND METHODS: Ten tissue samples removed by open prostatectomy were the source of tissue specimens. Two frozen sections were generated from each of several random biopsies taken from each adenoma immediately after enucleation, 1 of which was used for 5aR2 gene expression analysis and 1 for morphometric analysis. Results among biopsies were compared using the 5 alpha-reductase index (ratio of 5 alpha-reductase expression to an internal standard measured as electrophoretic band intensity). Morphometric composition was determined for smooth muscle, collagen, epithelium and glandular lumens. Statistical comparisons were performed with ANOVA by pairwise multiple comparison (Dunn) and Spearman's rank correlation procedure. RESULTS: For the 71 biopsies analyzed mean 5 alpha-reductase index was 0.23 +/- 0.16 and overall tissue distribution was smooth muscle 34%, collagen 35%, epithelium 14% and glandular lumens 17%. Inter-prostate and intraprostate variability in 5 alpha reductase index was statistically significant (p = 0.004) as was the variability in stromal-to-epithelial ratio (p = 0.012). The 5 alpha-reductase index showed strong correlation with stroma (%) and negative correlation with epithelium (%). CONCLUSIONS: Benign prostatic hyperplasia is heterogeneous in terms of tissue morphometry and expression of single important genes. This finding limits the use of single biopsy based markers to predict biological behavior, and has significant impact on the ability of distinguishing longitudinal changes in tissue composition from sampling artifacts. PMID- 12544311 TI - Validity and responsiveness of the national institutes of health chronic prostatitis symptom index. AB - PURPOSE: The National Institutes of Health Chronic Prostatitis Symptom Index (NIH CPSI) was validated in a sample of men with longstanding prostatitis but it has not been validated in primary care samples or assessed for responsiveness to change. We assessed its reliability, validity and responsiveness to change in a sample of men with pelvic pain visits to health maintenance organization primary care and urology clinics. MATERIALS AND METHODS: A total of 261 men with recent primary care or urology clinic visits for pelvic pain and no evidence of specific disease completed the NIH-CPSI and validated generic pain and health-related quality of life measures (Graded Chronic Pain Scale, Short-Form 36 and Short-Form 12 scales) in a telephone interview approximately 1 month after the visit, of whom 230 (88%) completed the measures again 3 months later. RESULTS: Validity was demonstrated for the NIH-CPSI pain and quality of life scales through their moderate correlations with other validated measures of these constructs. The NIH CPSI total scale but not the subscales showed high internal consistency. The pain, quality of life and total scores were moderately responsive to change (effect size -0.57 to -0.60), but the urinary symptoms scale was less responsive (effect size -0.18). CONCLUSIONS: The NIH-CPSI total score appears to be a valid, reliable, responsive measure of prostatitis symptoms in primary and secondary care patients. The findings support the use of the total score as an outcome measure. It may be useful to supplement it with valid, reliable measures of pain intensity and activity interference. PMID- 12544312 TI - Prostate biopsy culture findings of men with chronic pelvic pain syndrome do not differ from those of healthy controls. AB - PURPOSE: Previous reports have identified bacteria in the prostate of men with chronic pelvic pain syndrome. To examine whether prostatic bacteria are more prevalent among patients with chronic pelvic pain syndrome than among those without pelvic pain, we compared 4-glass urine test and prostate biopsy results. MATERIALS AND METHODS: A total of 120 patients with types IIIa and IIIb chronic pelvic pain syndrome and 60 asymptomatic controls underwent a standard 4-glass urine test, examination of expressed prostatic secretion leukocytes by hemocytometer and transperineal, digitally guided prostate biopsies. Tissue was cultured for aerobes, anaerobes, Trichomonas vaginalis, Chlamydia trachomatis and herpes simplex virus. Skin cultures were performed on a subset of patients and controls. RESULTS: Positive prostate biopsy cultures were obtained from patients and controls. Bacteria were found in 45 of 118 pain patients (38%) and in 21 of 59 controls (36%) (p = 0.74). Older men were more likely to have positive cultures. Men with type IIIa chronic pelvic pain syndrome were more likely than those with type IIIb to have positive prostate biopsy cultures. CONCLUSIONS: Bacteria cultured from transperineal prostatic biopsies do not differ between men with and without chronic pelvic pain syndrome. Prostatic bacteria obtained by biopsy are probably not etiologically related to the symptoms in the majority of men with chronic pelvic pain syndrome. PMID- 12544313 TI - The prevalence of men with National Institutes of Health category IV prostatitis and association with serum prostate specific antigen. AB - PURPOSE: We evaluated the prevalence and relationship of serum prostate specific antigen (PSA) levels in a screening population of men diagnosed with National Institutes of Health (NIH) category IV prostatitis. MATERIALS AND METHODS: In September of 2001, 300 men were randomly selected from our prostate cancer awareness screening program to be evaluated for NIH category IV prostatitis. After informed consent was obtained all patients completed the NIH prostate cancer awareness survey and had a serum sample obtained for PSA before examination. Expressed prostatic secretions were obtained from 227 of the 300 participants. Patients were classified according to findings on examination of the expressed prostatic secretions. The records were entered into our data base and subsequently reviewed. RESULTS: The prevalence of NIH category IV prostatitis was 32.2% in our population of men. Patient age, American Urological Association symptom scores and clinical prostate gland size did not differ between men with or without evidence of prostatitis on expressed prostatic secretion examination. Men with NIH category IV prostatitis had a mean serum PSA level of 2.3 which was significantly higher (p <0.0004) than those without prostatitis (mean PSA 1.4). CONCLUSIONS: These data suggest that NIH category IV prostatitis is fairly prevalent (32.2%) among men in the general population who present for prostate cancer screening and appears to contribute to increased serum PSA levels in some men. PMID- 12544314 TI - Terazosin therapy for chronic prostatitis/chronic pelvic pain syndrome: a randomized, placebo controlled trial. AB - PURPOSE: We evaluate terazosin therapy for chronic prostatitis/chronic pelvic pain syndrome. MATERIALS AND METHODS: The study included 100, 20 to-50-year-old subjects who met the consensus criteria for chronic prostatitis/chronic pelvic pain syndrome and had not received previous alpha-blockers. Subjects were randomized to receive terazosin with dose escalation from 1 to 5 mg. daily or placebo for 14 weeks. The primary criterion for response was scoring 2 or less ("delighted-to-mostly satisfied") on the National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI) quality of life item. The secondary criterion for response was greater than 50% reduction in NIH-CPSI pain score at 14 weeks. Other outcomes included total and NIH-CPSI domain scores, International Prostate Symptom Score, peak urinary flow rate, post-void residual urine and adverse effects. RESULTS: Using the primary criterion 24 of 43 evaluable subjects (56%) responded in the terazosin group compared to 14 of 43 (36%) in the placebo group (p = 0.03). Using the secondary criterion 26 of 43 subjects (60%) responded in the terazosin group compared to 16 of 43 (37%) in the placebo group (p = 0.03). The terazosin group had greater reductions (p <0.05) in NIH-CPSI total score, individual domain scores and International Prostate Symptom Score than the placebo group. There was no difference in peak urinary flow rate or post-void residual. In the terazosin group 18 patients (42%) had side effects compared to 9 (21%) in the placebo group (p = 0.04). CONCLUSIONS: Terazosin proved superior to placebo for patients with chronic prostatitis/chronic pelvic pain syndrome who had not received alpha-blockers previously. PMID- 12544315 TI - Editorial: Emerging concepts in the management of prostatitis/chronic pelvic pain syndrome. PMID- 12544316 TI - Corporeal plication for the treatment of congenital penile curvature. AB - PURPOSE: We evaluated the long-term outcome, effectiveness and patient satisfaction of corporeal plication for the correction of congenital penile curvature. MATERIALS AND METHODS: A total of 25 patients with congenital penile curvature were included in the study. Mean age was 39 years (range 15 to 45). Patients with Peyronie's disease, and/or chordee associated with hypospadias or evidence of erectile dysfunction were excluded from analysis. All procedures were done on an outpatient basis using local anesthesia. The technique of corporeal plication consists of placing longitudinal plication sutures of 2-zero braided polyester on the opposing side of curvature until it is corrected during artificially induced erection. A standardized questionnaire was then completed via telephone interview to assess results as well as the patient satisfaction rate. RESULTS: A total of 22 patients were available for evaluation. Successful results at up to 3(1/2) years (range 1 to 42 months, mean 18) of followup were achieved in 21 of the 22 patients (95%). Success was defined as the correction of curvature and a patient satisfaction rate of 75% or greater. Curvature was ventral in 20 cases, dorsal in 2 and left lateral in 3. Shortening of the penis after surgery was noted by 4 patients (18%), of whom 3 nevertheless had a satisfaction rate of 75% or greater. In 2 patients postoperative hematoma resolved spontaneously. Mean operative time was 50 minutes and 22 of the 25 patients (88%) received local anesthesia. CONCLUSIONS: Corporeal plication is a simple, minimally invasive surgical procedure that proved to be effective for congenital curvature of the penis compared with other, more extensive penile reconstruction surgery. PMID- 12544317 TI - Efficiency of percutaneous testicular sperm aspiration as a mode of sperm collection for intracytoplasmic sperm injection in nonobstructive azoospermia. AB - PURPOSE: We determined the feasibility of obtaining mature spermatozoa for intracytoplasmic sperm injection (ICSI) by percutaneous testicular sperm aspiration in men with nonobstructive azoospermia. We also compared the results of ICSI using spermatozoa recovered by open excisional biopsy versus percutaneous testicular sperm aspiration. MATERIALS AND METHODS: A total of 84 men with nonobstructive azoospermia underwent percutaneous testicular sperm aspiration to recover testicular spermatozoa for ICSI on the day of ova retrieval from the wife. Percutaneous testicular sperm aspiration was performed with the patient under general anesthesia in the upper and lower poles of each testis. It was followed by immediate microscopic search of the aspirate to confirm the presence of spermatozoa. In the absence of spermatozoa open excisional biopsy was performed in the same setting. RESULTS: Percutaneous testicular sperm aspiration resulted in the recovery of mature spermatozoa in 45 men (53.6%). Of the remaining 39 men (46.4%) requiring open biopsy adequate spermatozoa were recovered in 28 (71.8%). Although the fertilization rate was significantly higher in the sperm aspiration group, the cleavage and pregnancy rates were similar in the 2 groups. CONCLUSIONS: Percutaneous testicular sperm aspiration was a successful initial approach to collect mature spermatozoa in a high proportion of men with nonobstructive azoospermia. It is safe, minimally invasive and well tolerated by all patients. PMID- 12544318 TI - Tissue expander enhanced onlay island flap in the repair of severe hypospadias. PMID- 12544319 TI - Transrectal ultrasonography for the early diagnosis of adenocarcinoma of the prostate: a new maneuver designed to improve the differentiation of malignant and benign lesions. AB - PURPOSE: Transrectal ultrasound can reveal potentially malignant prostate lesions while they are still small. However, based on ultrasound alone they are often difficult to distinguish from benign focal lesions. We tested the reliability of a new technique for the sonographic evaluation of typical prostate lesions in differentiating adenocarcinoma from benign lesions. MATERIALS AND METHODS: During 18 months 398 consecutive male patients 45 to 76 years old underwent transrectal ultrasound for the early detection of prostate cancer. When suspicious hypoechoic lesions were noted in the peripheral regions of the prostate, moderate pressure was applied on the lesion using the ultrasound probe to evaluate consistency. Based on the response lesions were classified as deformable (the shape changed from approximately spherical to oval) or nondeformable (the original shape was retained). All lesions were then diagnosed based on fine needle biopsy. RESULTS: Peripheral hypoechoic prostate lesions were sonographically identified in 146 of 398 patients (36.7%). In 68 cases nondeformable lesions proved to be adenocarcinoma in 63 (92.6%), and chronic prostatitis and/or adenomatous hyperplasia in 5. In contrast, 62 of the 78 deformable nodules (79.5%) showed histological features of hyperplasia and/or chronic inflammation. The remaining 16 nodules, which showed more limited changes in shape during compression, were characterized by hyperplasia with acute inflammatory changes. In 5 cases there was also evidence of adenocarcinoma. CONCLUSIONS: Ultrasound guided compression of suspicious prostate lesions detected on transrectal sonography is a simple, rapid and reliable maneuver that may increase the diagnostic potential of this examination. PMID- 12544320 TI - Eccrine porocarcinoma of the penis. PMID- 12544321 TI - Rhabdomyolysis from cytochrome p-450 interaction of ketoconazole and simvastatin in prostate cancer. PMID- 12544322 TI - Spontaneous ruptured adrenal artery aneurysm. PMID- 12544323 TI - Repair of testicular artery injury during microsurgical varicocelectomy. PMID- 12544324 TI - Dermal graft repair of peyronie's disease complicated by epidermoid cyst. PMID- 12544325 TI - Herpes zoster following sacral nerve stimulation for overactive bladder. PMID- 12544326 TI - Re: Combined urinary and fecal diversion using a no bowel anastomosis technique. PMID- 12544327 TI - Re: Topography of the vanilloid receptor in the human bladder: more than just the nerve fibers. PMID- 12544328 TI - Hypercalcemia in a neonate with obstructive uropathy. PMID- 12544329 TI - The role of partial nephrectomy in the treatment of pediatric renal hypertension. AB - PURPOSE: From 5% to 25% of hypertension in children is renovascular compared with only 1% in adults. Although much attention is given to renovascular disease involving the main renal arteries, renin producing renal disease may also be intrarenal, involving abnormalities of the segmental vessels or renal parenchyma. We present our results of partial nephrectomy in this unique group of pediatric patients with hypertension in whom renin dependent hypertension involved only a segment of the kidney. MATERIALS AND METHODS: Six patients 10 months to 16 years old were referred for the evaluation of hypertension. Initial evaluations included computerized tomography, ultrasound, voiding cystourethrography and radionuclide renal scan. Renal arteriography with renal vein renin sampling was performed in 5 patients, of whom 4 underwent selective segmental renal vein sampling. Diagnoses included segmental hypoplasia (Ask-Upmark kidney), reflux associated scarring and renal arteriovenous malformation. RESULTS: In patients who underwent selective segmental renal vein sampling an increased renin level was present in the area of the renal lesion (mean 24.9 ng./ml. per hour, range 9.2 to 40.6) compared to the ipsilateral renal vein (15.6, 114 to 29.8). Three patients had evidence of contralateral suppression of renin secretion. All 6 patients underwent upper, lower or mid segment partial nephrectomy. All patients became immediately normotensive and remained so at a mean 10 years of followup. CONCLUSIONS: Partial nephrectomy provides an excellent nephron sparing cure for segmental renal hypertension. In the pediatric population selective segmental renal vein renin sampling is invaluable for locating the renin producing lesion. As in adults, contralateral renin suppression is predictive of surgical cure. PMID- 12544330 TI - Pediatric staghorn calculi: the role of extracorporeal shock wave lithotripsy monotherapy with special reference to ureteral stenting. AB - PURPOSE: Treatment for staghorn calculi in children represents a unique challenge. We assessed the efficacy of extracorporeal shock wave lithotripsy (ESWL) (Dornier Medical Systems, Inc., Marietta, Georgia) monotherapy for the management of staghorn calculi in children with special reference to ureteral stenting. MATERIALS AND METHODS: From June 1992 to January 2001 we treated 42 children 9 months to 12 years old with staghorn stones using the Piezolith 2501 (Richard Wolf GmBH, Knittlingen, Germany) lithotriptor. The initial group of 19 patients underwent ESWL without prophylactic ureteral stenting, while in the latter group of 23 a Double-J (Medical Engineering Corp., New York, New York) ureteral stent was inserted immediately before the first ESWL session. Mean patient age, stone size, number of shock waves and ESWL sessions, hospital stay, stone-free rate and major complications were compared in the 2 groups. RESULTS: Overall 33 children (79%) were stone-free after 3 months. The 2 groups were comparable in regard to patient age, stone size, number of shock waves and ESWL sessions, and stone-free rates. Major complications developed in 21% of the unstented group, whereas none were observed in stented cases. This difference was statistically significant (p = 0.035). Seven post-ESWL auxiliary procedures were required in the unstented group to manage complications. Hospital stay was significantly longer in the unstented compared with the stented group (p = 0.022). At a followup of 9 to 102 months (mean 47) stones recurred in 2 children, who were treated with further ESWL. CONCLUSIONS: ESWL monotherapy was an efficient and safe modality for the treatment of staghorn calculi in children. Stented patients had fewer major complications and a shorter hospital stay. Prophylactic ureteral stenting is advisable before ESWL for staghorn calculi in children. PMID- 12544331 TI - Management of pediatric urolithiasis in Pakistan: experience with 1,440 children. AB - PURPOSE: We evaluated the efficacy and safety of different modalities for pediatric urolithiasis in a developing country in 2 eras, namely before and after the advent of minimally invasive surgery. MATERIALS AND METHODS: We retrospectively reviewed the records of 1,440 children younger than 14 years treated with various modalities during a 14-year period. From 1987 to 1995, 486 and 50 patients were treated with open surgery, and extracorporeal shock wave lithotripsy (ESWL, Dornier Medical Systems, Inc., Marietta, Georgia) and minimally invasive methods, respectively. Between 1996 and 2000, 518 and 386 children were treated with surgery and minimally invasive methods, respectively. RESULTS: Of the 1,440 children 795 (55.2%) had renal, 198 (13.8%) had ureteral and 447 (31%) had bladder calculi. Of the renal stones 556 (70%), 177 (22%) and 62 (7.8%) were treated with open surgery, ESWL and percutaneous nephrolithotomy, respectively. Of the ureteral calculi 85 (43%), 37 (18.6%) and 76 (38%) were managed by ESWL, ureterorenoscopy and open surgery, respectively. Of the bladder calculi 307 (68%), 77 (17.2%) and 63 (14%) were treated with open vesicolithotomy, transurethral pneumatic cystolithotripsy and ESWL, respectively. The renal stone clearance rate was 98% after open surgery, 84% after ESWL and 68% after percutaneous nephrolithotomy monotherapy at 3 months of followup. Similarly the ureteral stone-free rate was 54% after ESWL and 86.9% after ureterorenoscopy. Of the patients with bladder calculi 48% and 93% become stone-free after ESWL and transurethral pneumatic cystolithotripsy, respectively. CONCLUSIONS: The use of ESWL, percutaneous nephrolithotomy and ureterorenoscopy has resulted in treating a large number of children with a short hospital stay and early return to school. Open surgery is reserved only for complex stones. PMID- 12544332 TI - Comparison of laparoscopic versus open partial nephrectomy in a pediatric series. AB - PURPOSE: Recent advances in laparoscopic surgery as well as increasing experience with these techniques have led to the selection of laparoscopic surgery for many urological procedures. A lesser number of pediatric laparoscopic surgical studies have been reported. Few pediatric comparative laparoscopic versus open surgical procedure studies have been published. We compared 2 groups of similar pediatric patients who underwent partial nephrectomy via the laparoscopic or open technique. MATERIALS AND METHODS: A total of 22 consecutive partial nephrectomies were performed in pediatric patients 3 months to 15 years old. Of these procedures 11 chosen according to surgeon preference were performed laparoscopically and 11 were done by the open technique. Clinical data were obtained by chart review and compared retrospectively in the 2 groups. Demographic data, operative time and blood loss, the perioperative complication rate, hospital stay and costs, postoperative analgesic use and followup findings were compared. RESULTS: Mean operative time in the laparoscopic and open groups was 200.4 and 113.5 minutes, respectively (p <0.0005). Blood loss was less than 50 cc in all patients. In the laparoscopic and open groups mean hospital stay was 25.5 and 32.6 hours (p = 0.068), and mean cost was $6,125 and $4,244 (p = 0.016), respectively. Patients in the laparoscopic group required fewer doses of analgesics than those who underwent open surgery (mean 10.9 versus 21, p = 0.041). CONCLUSIONS: Our findings show that increased operative time and costs are disadvantages of pediatric laparoscopic nephrectomy compared with open techniques. Conversely decreased hospital stay, lower analgesic requirements and cosmesis support the use of laparoscopy for pediatric partial nephrectomy. These differences must be considered when deciding which technique is best for overall patient care. PMID- 12544333 TI - Functional evaluation by quantitative dimercaptosuccinic Acid scintigraphy after kidney trauma in children. AB - PURPOSE: Most pediatric surgical teams have adopted nonoperative treatment for a traumatic kidney lesion in children. In the emergency setting and at long-term followup dimercaptosuccinic acid (DMSA) scintigraphy enables us to identify the consequences on global renal function as well as on the function of each individual kidney. MATERIALS AND METHODS: Eight boys and 12 girls between 0.6 and 15.9 years old (average age 9.7) were evaluated after including renal trauma, minor and major injury in 10 each. Scintigraphy was performed 8 days and 6 months after injury. The tracer used was 2 MBq./kg. Tc-DMSA. Semiquantitative analysis of the images consisted of determining 2 parameters, namely relative renal function, corresponding to the fraction of activity of a single kidney compared to the activity of the 2 kidneys, and the calculated renal activity fraction, corresponding to the function of each kidney compared with the activity of a theoretical kidney in a child of the same age. The posttraumatic renal scintigraphy series was paired with a series of healthy children matched by age, weight, height, sex and affected side of function as a control group. Global renal function was also measured using a formula based on ethylenediaminetetraacetic acid scintigraphy with the height, weight and increased serum creatinine in each patient. RESULTS: For minor injuries the quantitative functional evaluation revealed a significant average loss of renal function +/- SD on the side of the lesion (12.8% +/- 3.1% versus 18.3% +/- 2%, p = 0.001). For major injuries on relative renal function evaluation we noted an average 36-point difference in the damaged and contralateral kidneys 8 days after the accident. Six months after trauma we noted a definitive loss of 48% in the calculated renal activity fraction on the side of the lesion. There was no compensatory hypertrophy on the noninjured side when the calculated renal activity fraction was compared with that of a normal kidney in a control patient (mean 19.1% +/- 4.2% versus 19.5% +/- 3.7%, not significant). Creatinine clearance was normal in each patient. DISCUSSION A renal contusion always induces parenchymal loss. Major kidney trauma has significant consequences on the opposite side. At 6 months the presumably uninjured contralateral kidney may be limited in its ability to compensate through hypertrophy, which worsens the global renal functional prognosis of a traumatic but initially unilateral lesion. CONCLUSIONS: Posttraumatic functional evaluation by DMSA scintigraphy, which measures the nephron capital of each kidney and the 2 together, seems essential to inform patients about the seriousness of the lesion and lead them to an eventual long-term nephrological followup with regular blood pressure assessment. PMID- 12544334 TI - Spontaneous regression of cystic dysplasia of the testis. PMID- 12544335 TI - Males with Down's syndrome and nonneurogenic neurogenic bladder. AB - PURPOSE: We assessed the relationship of trisomy 21 with the most severe dysfunctional elimination syndrome, nonneurogenic neurogenic bladder or the Hinman-Allen syndrome. MATERIALS AND METHODS: We retrospectively reviewed our experience with children with Down's syndrome (trisomy 21) in a 10-year period and identified a subset who presented for the evaluation and treatment of urinary tract infections associated with severe disorders of urinary and fecal elimination. RESULTS: Four males 9 months, 14 years, 18 years and 21 years old met the criteria for review. All patients underwent radiological and urodynamic evaluation, and were diagnosed with hydronephrosis and prostatic urethral dilatation with pelvic floor spasticity. Renal function studies showed a creatinine of 0.7 mg./dl. in the 9-month-old, 1.2 mg./dl. in the 14-year-old, 1.9 mg./dl. in the 18-year-old and 2.2 mg./dl. in the 21-year-old patient. Three patients underwent surgical treatment to protect the upper urinary tract, including bladder augmentation cystoplasty and an appendiceal Mitrofanoff stoma in 2, and vesicostomy in 1. The remaining patient was treated conservatively with a behavioral modification program that included timed voiding and a bowel regimen. CONCLUSIONS: Boys with trisomy 21 may be at risk for the Hinman-Allen syndrome. Surgical intervention and clean intermittent catheterization for renal preservation and continence can be performed in this population despite intellectual impairment. Further evaluation is necessary to determine whether this relationship is more common than appreciated and whether this syndrome occurs in females with trisomy 21. PMID- 12544336 TI - The Indiana experience with artificial urinary sphincters in children and young adults. AB - PURPOSE: We reviewed a 22-year single institutional experience with the artificial urinary sphincter in children and adolescents. To our knowledge this report represents the largest series in the world in children. MATERIALS AND METHODS: Between 1980 and 2002, 142 patients underwent implantation of an artificial urinary sphincter, of whom 93 males and 41 females with a median age of 10 years (range 3 to 39) were available for analysis. A total of 59 patients initially received an AMS 742/792 (American Medical Systems, Inc., Minnetonka, Minnesota) artificial urinary sphincter, of whom 33 were subsequently changed to an AMS 800, while 75 initially received an AMS 800 model. Sphincter followup was terminated at device removal or at the last documented contact. The etiology of incontinence was neuropathic bladder in 107 cases (80%), the exstrophy/epispadias complex in 21 (16%) and other in 6 (4%). Outcome measures included continence, mechanical complications (leakage, tube kink and pump malfunction), functioning sphincter revisions (change in cuff size, pump repositioning and bulbar cuff placement), surgical complications (erosion, infection and misplacement) and associated surgical procedures. Mean followup of the pre-800 and 800 models was 6.9 (range 0.2 to 21.5) and 7.5 years (range 0.1 to 17.1), respectively. Fisher's exact test, Kaplan-Meier life analysis and the chi-square test were used for statistical analysis. RESULTS: After artificial urinary sphincter placement in the 134 patients continence was achieved in 86%, improved in 4% and not achieved in 10%. Of those with a sphincter in place 92% were continent. In terms of bladder emptying after artificial urinary sphincter insertion 22% of patients voided, 11% voided combined with clean intermittent catheterization, 48% performed clean intermittent catheterization only via the urethra, 16% performed it via a catherizable channel and 3% used urinary diversion. A mechanical complication developed in 38 of the 59 patients (64%) with pre-800 model compared with 33 of the 109 (30%) with the 800 model (p <0.0001). A mechanical complication occurred every 7.6 versus 16 patient-years for the pre-800 versus 800 models (p = 0.0001). Revision was required in 15 of the 59 patients (25%) with a pre-800 model versus 17 of the 109 (16%) with the 800 model (p = 0.103). Revision was performed every 22.7 versus 44.3 patient-years for the pre-800 versus the 800 model (p = 0.023). The artificial urinary sphincter eroded in 11 of the 59 patients (19%) with the pre-800 versus 17 of the 109 (16%) with the 800 model (p = 0.52). Ten patients experienced a total of 12 perforations of the augmented bladder after artificial urinary sphincter implantation. A total of 164 secondary surgical procedures were performed, including 38 of 134 bladder augmentations (28%). A total of 30 sphincters were permanently removed. CONCLUSIONS: The artificial urinary sphincter is the only bladder neck procedure that allows spontaneous voiding in the neuropathic population, obviates the need for clean intermittent catheterization and yet is compatible with it when necessary. It is also equally versatile in the 2 genders. Mechanical complications occur but they were dramatically decreased by the modifications of the AMS 800 model. In addition, secondary bladder augmentation was required in 28% of our patients. Lifelong followup is mandatory in all patients with an artificial urinary sphincter. PMID- 12544337 TI - Diameter of the external urethral sphincter as a predictor of detrusor-sphincter incoordination in children: comparative study of voiding cystourethrography. AB - PURPOSE: Voiding cystourethrography is a diagnostic procedure widely used to evaluate lower urinary tract abnormalities in children. In children with and without suspected voiding dysfunction we measured the internal diameter of the external urethral sphincter on voiding cystourethrography to evaluate its diagnostic accuracy as a predictor of detrusor-sphincter incoordination. MATERIALS AND METHODS: In part 1 of the study 59 boys and 37 girls with normal voiding function underwent voiding cystourethrography. In each child 1 to 6 voiding cystourethrograms (mean 2.1) were performed as a part of urological evaluation. Thus, a total of 200 voiding cystourethrograms were obtained. Underlying urological disease was primary vesicoureteral reflux in 57 cases, congenital hydronephrosis in 9, urinary tract infection in 6, ureteral anomalies in 11 and other in 13. To determine external urethral sphincter inner diameter on a given voiding cystourethrogram the sphincter was measured on consecutive images and the widest diameter was chosen. In part 2 of the study 43 children with suspected voiding dysfunction underwent voiding cystourethrography and external urethral sphincter electromyography. Electromyography results were comparatively analyzed with external urethral sphincter data. RESULTS: In part 1 an age dependent increase in external urethral sphincter inner diameter was noted in children with normal voiding function. External urethral sphincter inner diameter in mm. was determined by the formula, 0.166 x age in years + 4.31 in boys (p = 0.0001, r = 0.374) and 0.222 x age in years + 2.73 in girls (p <0.0001, r = 0.595). Overall the incidence of an external urethral sphincter inner diameter of less than 3 mm. was only 4% (8 of 200 patients). In part 2 external urethral sphincter electromyography documented detrusor-sphincter incoordination in 15 children (35%). When we defined an external urethral sphincter inner diameter of less than 3 mm. as a cutoff value for predicting detrusor-sphincter incoordination, the sensitivity, specificity, positive and negative predictive values were 93%, 89%, 82% and 96%, respectively. CONCLUSIONS: External urethral sphincter inner diameter has satisfactory sensitivity and specificity as a predictor of detrusor-sphincter incoordination in children. Thus, simple measurement of the inner diameter of the external urethral sphincter on voiding cystourethrography should be recommended in children with suspected voiding dysfunction before performing rather invasive urodynamic studies. PMID- 12544338 TI - Testis histopathology in boys with cryptorchidism correlates with future fertility potential. AB - PURPOSE: We studied the ability of testis biopsy to predict eventual fertility potential. MATERIALS AND METHODS: A total of 25 patients with unilateral undescended testis and 11 with bilateral undescended testes who previously underwent orchiopexy at a median age of 9 years were followed until after age 18 years. Semen analysis, testicular volume, follicle-stimulating hormone, luteinizing hormone, testosterone and inhibin B were compared to previously obtained biopsies of each testis. Biopsies were graded as mild, moderate or severe histology based on published data for normal and undescended testis histology. RESULTS: Patients with unilateral undescended testis and moderate histology were more likely to have a sperm density of greater than 20 million per cc than those with severe histology (p = 0.006), although no difference in hormonal parameters was noted. Patients with bilateral undescended testes and moderate histology were more likely to have normal follicle-stimulating hormone and inhibin B than those with severe histology (p = 0.05 and 0.002, respectively), although no difference in semen analysis was observed. Overall 7 of the 9 patients with bilateral undescended testes had less than 20 million sperm per cc. CONCLUSIONS: Testis biopsy provides useful prognostic information about semen analysis in patients with unilateral undescended testis. In the 9 patients with bilateral undescended testes testis biopsy divided them into normal and abnormal groups in terms of hormonal parameters. Testis biopsy was not able to provide additional prognostic information on semen analysis in patients with bilateral undescended testes since they were uniformly poor. PMID- 12544339 TI - Testicular torsion: direction, degree, duration and disinformation. AB - PURPOSE: We reviewed and contrast with the literature the cumulative clinical experience at our pediatric urological division in the last 20 years with managing testicular torsion, focusing specifically on the direction and degree of testicular torsion and the duration of symptoms before presentation. We also addressed the incidence of gastrointestinal symptoms, role of manual detorsion, residual torsion and incidence of atrophy. MATERIALS AND METHODS: We reviewed the medical records of 200 consecutive males 18 months to 20 years old who underwent surgical exploration by a pediatric urologist for a diagnosis of testicular torsion between 1980 and 2000. RESULTS: Of 186 nonelective explorations symptoms were localized to the left side in 52% and to the right side in 48%. Information on the direction and degree of testicular rotation was available in 162 of 186 cases (87%) and anticipated medial rotation occurred in only 108 (67%). Lateral rotation in 54 of 162 cases (33%) occurred in 28 of 84 (33%) with left torsion and in 26 of 78 (33%) with right torsion. A median of 540 degrees of torsion (range 180 to 1,080) was noted in the 70 orchiectomy cases (38%) and a median of 360 degrees (range 180 to 1,080) was noted in the 116 salvaged testes (62%). Manual detorsion was attempted in 53 orchiopexy cases with residual torsion in 17 (32%). Testicular atrophy developed in 27% of the patients. CONCLUSIONS: The traditional teaching that testicular torsion occurs primarily in the medial direction is misleading since in a third of cases it occurs in the lateral direction. While manual detorsion should be guided by response and return of normal anatomy, surgical exploration remains necessary since residual torsion still poses a risk to testicular viability. Long-term followup is warranted to assess the true incidence of subsequent atrophy after the management of acute testicular torsion. PMID- 12544340 TI - Optimizing the operative treatment of boys with varicocele: sequential comparison of 4 techniques. AB - PURPOSE: We compared 4 techniques of varicocele ligation in boys and young adolescents to determine the optimal operative treatment that avoids varicocele recurrence and postoperative hydrocele formation. MATERIALS AND METHODS: In 10 years a total of 128 varicocelectomies were performed sequentially in 121 boys and young adolescents with a mean age of 12 years using the laparoscopic, inguinal testicular artery sparing, standard Palomo (high mass retroperitoneal ligation) and modified Palomo approaches. The modified Palomo approach involved suprainguinal and retroperitoneal ligation of the veins and artery, and microsurgical sparing of the blue stained lymphatic pathway of the testis. Patients were followed a mean of 52 months. RESULTS: In the 19 boys in the laparoscopy group varicocele persisted in 10% and hydrocele developed in 5%. In the 21 patients who underwent inguinal surgery with artery preservation recurrent varicoceles were identified in 14% and no hydroceles were observed. In the 32 patients who underwent the standard Palomo procedure there was no palpable varicocele persistence or recurrence, while hydroceles developed in 12%. Of the 56 patients in the modified Palomo group varicocele recurred in 1 (2%) and there were no hydroceles. No testicular atrophy developed in any patient. CONCLUSIONS: Comparison of all 4 groups revealed significant differences in varicocele recurrence (p = 0.038) and hydrocele formation (p = 0.023). Pairwise group comparison showed that the modified Palomo technique resulted in a significant decrease in the incidence of postoperative hydrocele formation compared with the standard Palomo method (p = 0.015). This procedure can be recommended as the optimal surgical technique for varicocele treatment in males of this young age. PMID- 12544341 TI - Fibroblast growth factors and their receptors in transitional cell carcinoma. AB - PURPOSE: The recent identification in transitional cell carcinoma of mutations in a fibroblast growth factor (FGF) receptor, namely FGF receptor-3, has provoked great interest in the potential usefulness of FGF receptors and their ligands as molecular markers and as targets for bladder cancer therapy. We examined these possibilities in light of the published literature. MATERIALS AND METHODS: We reviewed the world literature on FGFs and their receptors from 1966 to January 2002 using PubMed. RESULTS: The recent identification in transitional cell carcinoma of a high frequency of mutations in FGF receptor-3 predicted to activate kinase activity of the receptor indicate a likely role as an oncogene in the urothelium. The finding of FGF receptor-3 mutations only rarely in other tumor types to date indicates surprising urothelial specificity that requires tissue specific approaches for evaluation and exploitation. In contrast, FGF receptor-2 expression is down-regulated in bladder tumors, suggesting a possible tumor suppressor role. Information is available on the expression of FGF receptors-1 and 2 in normal bladder and urine, and in bladder tumors. These angiogenic factors represent potential urine markers of bladder neoplasia, although as single markers they lack sufficient sensitivity and specificity. Some interesting insights into the potential role of these factors have come from studies using in vitro model systems. However, there is little information on the numerous other members of this family of growth factors in the bladder and, therefore, much scope for future studies. CONCLUSIONS: It is clear that the FGFs and their receptors have important roles in the development of transitional cell carcinoma. Undoubtedly it will be a focus for much future research. It can be anticipated that members of these protein families would represent useful clinical markers and potential targets for bladder cancer therapy. PMID- 12544342 TI - Hepatic alanine-glyoxylate aminotransferase activity and oxalate metabolism in vitamin B6 deficient rats. AB - PURPOSE: Urinary oxalate has an important role in the formation of calcium oxalate stone and approximately 50% to 60% of urinary oxalate is derived from the endogenous metabolism of glyoxylate. Glyoxylate is enzymatically converted to glycine by alanine-glyoxylate aminotransferase in the liver and vitamin B6 has a key role as a coenzyme. Therefore, we evaluated hepatic alanine-glyoxylate aminotransferase activity and oxalate metabolism in vitamin B6 deficient rats. MATERIALS AND METHODS: A total of 12 male Wistar rats were fed a control or a vitamin B6 deficient diet. After 4 weeks creatinine, oxalate, glycolate, glycine and citrate in the urine, and alanine-glyoxylate aminotransferase activity and its mRNA level in the liver were measured. RESULTS: Urinary oxalate-to-creatinine and glycolate-to-creatinine ratios were significantly higher in vitamin B6 deficient rats than in control rats but urinary glycine-to-creatinine and citrate to-creatinine ratios were significantly lower. Hepatic alanine-glyoxylate aminotransferase activity and its mRNA level were significantly lower in vitamin B6 deficient rats than in control rats. CONCLUSIONS: Vitamin B6 deficiency not only decreased alanine-glyoxylate aminotransferase activity, but also down regulated alanine-glyoxylate aminotransferase gene expression by hepatocytes and led to hyperoxaluria and hyperglycolic aciduria secondary to impaired metabolism of oxalate precursors. Hyperoxaluria with hypocitruria may also contribute to calcium oxalate stone formation in vitamin B6 deficiency. PMID- 12544343 TI - Reference range for gastrointestinal oxalate absorption measured with a standardized [13C2]oxalate absorption test. AB - PURPOSE: Hyperoxaluria is a prominent risk factor for calcium oxalate urinary stones. Oxalate in urine is synthesized in the body or absorbed from food in the gastrointestinal tract. The amount of oxalate absorbed by patients with calcium oxalate stones may vary from a few percent to 50% of the dietary intake. Reference values for oxalate absorption measured under a standardized diet have never been attained in sufficient numbers from healthy individuals. Therefore, to our knowledge we collected for the first time the values required to interpret test results in patients with recurrent urinary stones. MATERIALS AND METHODS: A total of 120 healthy volunteers, including 60 females and 60 males, received an identical standard diet on 2 consecutive days. On the morning of day 2 a capsule containing 0.37 mmol. sodium [13C2]oxalate (not radioactive) was ingested with water. Urinary oxalate was measured by gas chromatography-mass spectrometry. Absorption at a fixed 800 mg. daily Ca input is expressed as a percent of the labeled oxalate dose. RESULTS: For the standardized [13C2]oxalate absorption test the reference range in 95% of the 120 volunteers was 2.2% to 18.5% (mean +/- SD 7.9% +/- 4.0%). The repeatability of the standardized test was determined in 26 of the 120 volunteers by repeating the test twice. The mean intra-individual SD was 3.39% +/- 1.68%. CONCLUSIONS: We assessed reference values of intestinal oxalate absorption using a standardized diet. Interindividual and intra individual variance was high. PMID- 12544344 TI - Laparoscopic anatrophic nephrolithotomy: feasibility study in a chronic porcine model. AB - PURPOSE Anatrophic nephrolithotomy performed via open surgery involves incising the renal parenchyma along an avascular plane to remove a large, complex renal stone. We determined the feasibility of performing laparoscopic anatrophic nephrolithotomy in a survival porcine model. Furthermore, we present a novel technique of creating a staghorn calculus in the porcine model. MATERIALS AND METHODS After developing the technique in 3 pigs the survival study was performed in 10 consecutive animals. The procedure comprised 2 aspects. 1) We developed an animal model for staghorn calculi by retrograde injection of polyurethane (Fomo Products, Inc., Norton, Ohio) into the renal pelvis through a ureteral catheter. For a 2-week period the staghorn calculus was allowed to create hydronephrosis. 2) Laparoscopic anatrophic nephrolithotomy was done, involving control of the renal artery and vein, in situ renal hypothermia with ice slush in 1 animal, lateral renal parenchymal incision, stone extraction and suture repair of the incised collecting system and renal parenchyma. RESULTS Synthetic stone formation and laparoscopic anatrophic nephrolithotomy were successful in all 10 animals, including 1 that underwent staged bilateral anatrophic nephrolithotomy. Mean operative time for anatrophic nephrolithotomy was 125 minutes. Mean blood loss was 68 cc and mean warm ischemia time was 30 minutes (range 23 to 39). A residual small pelvicaliceal calculus was noted postoperatively in the initial 3 cases only. Thereafter, routine intraoperative ultrasonography and flexible endoscopy were done for stone localization, resulting in a stone-free rate of 100% in all 7 remaining animals. Diethylenetriamine pentaacetic acid renal scans documented improvement in the glomerular filtration rate from a mean of 26.4 ml. per minute after stone creation and hydronephrosis to 54.8 ml. per minute 4 to 5 weeks after laparoscopic anatrophic nephrolithotomy. CONCLUSIONS Laparoscopic techniques can be applied to complex stone surgery such as anatrophic nephrolithotomy with encouraging surgical and functional outcomes. To our knowledge this report represents the initial study of in situ creation of experimental staghorn calculi and laparoscopic anatrophic nephrolithotomy performed completely intracorporeally in a chronic porcine model. PMID- 12544345 TI - Evaluation of a vessel sealing system, bipolar electrosurgery, harmonic scalpel, titanium clips, endoscopic gastrointestinal anastomosis vascular staples and sutures for arterial and venous ligation in a porcine model. AB - PURPOSE: We assessed the usefulness of the LigaSure (Valleylab, Boulder, Colorado) vessel sealing system for vascular control during laparoscopic surgery and compared it with other available hemostatic modalities. MATERIALS AND METHODS: A total of 31 domestic pigs were divided into 5 groups. In groups 1 and 2 the vessel sealing system was compared with titanium clips and Endo-GIA (United States Surgical, Stamford, Connecticut) staples. In group 3 the vessel sealing system was compared with standard Klepinger (Karl Storz, Culver City, California) bipolar forceps. In group 4 the harmonic scalpel and Trimax (United States Surgical) bipolar forceps were compared. In group 5 in vivo laparoscopic application of the vessel sealing system was evaluated. RESULTS: The 5 mm. laparoscopic vessel sealing system sealed arteries up to 6 mm. and veins up to 12 mm. in diameter at supraphysiological bursting pressure. We evaluated 13 arteries with a diameter of 6 mm. or less at a mean bursting pressure of 662 mm. Hg (range 363 to 1,985) and 11 veins with a diameter of 12 mm. or less with a mean bursting pressure of 233 mm. Hg (range 63 to 440). Collateral tissue damage extended 1 to 3 mm. from the application site. Standard bipolar energy with Klepinger and Trimax forceps was less reliable and in some cases vessel sealing could not be accurately assessed before vessel division. Collateral tissue injury was 1 to 6 mm. The harmonic scalpel did not reliably seal vessels larger than 3 mm. but resulted in the least acute collateral tissue injury of 0 to 1 mm. CONCLUSIONS: In the porcine model the LigaSure system is a viable option for laparoscopic management of arteries up to 6 mm. and veins up to 12 mm. in diameter. PMID- 12544346 TI - Isolation of circulating cancer cells from whole blood by immunomagnetic cell enrichment and unenriched immunocytochemistry in vitro. AB - PURPOSE: We improved tumor cell detection compared with currently available immunocytochemical methods by immunomagnetic cell enrichment. MATERIALS AND METHODS: Two methods of immunomagnetic enrichment using antibody coated magnetic beads were tested and compared with unenriched immunocytochemistry, including positive selection of epithelial cells with the antiepithelial antibody BER-EP4 and depletion of mononuclear cells with the anti-leukocyte antibody CD45. Various numbers of tumor cells from the 4 tissue culture cell lines DU 145, RT-4, KTCTL-2 and KTCTL-30 obtained from urological tumors were added to whole blood and mononuclear cells were isolated by density centrifugation. After incubation of the cell suspensions with beads cell separation was done in a magnetic field. After centrifugation on glass slides immunocytochemical staining for cytokeratin was performed. A total of 96 experiments were completed and negative controls were obtained. RESULTS: The number of tumor cells detected by positive selection and depletion was significantly higher compared with immunocytochemistry (Wilcoxon test p <0.01). Mean enrichment factor and tumor cell recovery rates were 12.9% and 43.5% for positive selection, and 9.4% and 32.6% for depletion, respectively (p <0.05). With 1 tumor cell suspended in up to 30 ml. full blood unenriched immunocytochemistry failed to detect cancer cells, whereas positive selection revealed epithelial cells in 12 of 14 cases (85.5%) and depletion in all 14 (p <0.05). No false-positive results were observed. CONCLUSIONS: Compared with unenriched immunocytochemistry immunomagnetic enrichment significantly improves the detection of epithelial cells added to blood. A significant advantage was observed for positive selection. Immunomagnetic enrichment may be important for clinical practice in the future. PMID- 12544347 TI - Significance of thymidine kinase activity in renal cell carcinoma. AB - PURPOSE: Thymidine kinase (TK) has a key role in the complementary or alternative salvage pathway of pyrimidine synthesis. Little is known about the significance of TK activity in renal cell carcinoma. We examined the activity of TK in 66 renal cell carcinomas and investigated the association of the level of TK activity with the stage and grade of renal cell carcinoma. MATERIALS AND METHODS: TK activity in nonfixed, fresh frozen renal cell carcinoma and normal kidney was determined by the DEAE cellulose disc method. RESULTS: The activity of TK was approximately 4-fold higher in normal kidney compared with renal cell carcinoma. The higher the stage and grade of renal cell carcinoma, the higher the TK activity. TK activity correlated positively with the activity of thymidine synthase, which is a key enzyme for pyrimidine synthesis in the de novo pathway. TK activity in renal cell carcinoma cells was inversely related to their sensitivity to 5-fluorouracil. CONCLUSIONS: To our knowledge this report represents the first study to demonstrate that the level of TK activity correlates with the stage and grade of renal cell carcinoma, and higher TK activity in renal cell carcinoma predicts lower sensitivity to 5-fluorouracil. These results suggest that high TK activity may be associated with the malignant potential of renal cell carcinoma and TK may be a molecular therapeutic target for this disease. In addition, the combination of TK and thymidylate synthase inhibitors may be beneficial for renal cell carcinoma. PMID- 12544348 TI - Elevated Akt activation and its impact on clinicopathological features of renal cell carcinoma. AB - PURPOSE: Akt has been implicated in the pathogenesis and progression of numerous human malignant tumors because Akt regulates many key effector molecules involved in controlling the balance of survival and apoptosis. Elevated Akt activation has been demonstrated in various types of malignant tumors. In the current study we evaluated Akt activation in renal cell carcinoma and investigated its association with pathological features and clinical outcome. MATERIALS AND METHODS: Akt activation was evaluated by immunohistochemistry using antiphospho-specific Akt antibody, which recognizes only activated Akt, in 48 patients with renal cell carcinoma. High immunostaining tumors were defined as tumors with increased staining intensity compared with adjacent nonneoplastic kidney tissue and low immunostaining tumors were defined as tumors with equivalent or decreased staining intensity. The association of Akt activation status with clinicopathological features was analyzed. RESULTS: Of 48 patients examined 18 (37.5%) demonstrated high antiphospho-specific Akt immunostaining compared with adjacent nonneoplastic kidney tissue, while 30 (62.5%) demonstrated low immunostaining. Elevated immunostaining was significantly associated with tumor grade (p = 0.0354) and metastatic disease (p = 0.0044), while it was not associated with tumor stage or histological subtype. In addition, high antiphospho-specific Akt immunostaining was significantly associated with a poor cancer specific survival rate on univariate analysis (p = 0.0109) but not on multivariate analysis. CONCLUSIONS: Elevated Akt activation could be a common finding, especially in high grade tumor and metastatic disease. It could have an important role in the pathogenesis and progression of renal cell carcinoma. PMID- 12544349 TI - Plasma levels of insulin-like growth factor-1 and binding protein-3, and their association with bladder cancer risk. AB - PURPOSE: Because insulin-like growth factors (IGFs) and their binding proteins have been implicated in the development of prostate, breast, colon and lung cancer, we examined the role of IGF-1 and IGF binding protein-3 levels in bladder cancer risk. MATERIALS AND METHODS: We used an enzyme-linked immunosorbent assay to compare plasma levels of IGF-1 and IGF binding protein-3 in 154 patients with bladder cancer and 154 controls from an ongoing case-control study. RESULTS: Mean IGF-1 was significantly higher in cases than in controls (175.8 versus 153.2 ng./ml., p <0.01). Mean IGF binding protein-3 was significantly lower in cases than in controls (2,632.9 versus 3,056.6 ng./ml., p <0.01). The highest quartile plasma levels of IGF-1 were associated with an increased risk of bladder cancer (OR 3.10, 95% CI 1.43 to 6.70) and the highest quartile plasma levels of IGF binding protein-3 were associated with a reduced risk of bladder cancer (OR 0.38, 95% CI 0.19 to 0.78). The effects were more striking when IGF-1 and IGF binding protein-3 levels were analyzed together. In addition, a higher molar ratio of IGF 1-to-IGF binding protein-3 was associated with an increased risk of bladder cancer (OR 4.30, 95% CI 1.99 to 9.28). Dose-response relationships were evident when subjects were categorized into quartiles by the values of IGF-1, IGF binding protein-3 and the molar ratio in controls. CONCLUSIONS: To our knowledge this is the first study to suggest that patients with bladder cancer have higher plasma levels of IGF-1 and lower levels of IGF binding protein-3 than controls. Thus, measuring plasma IGF-1 and IGF binding protein-3 may be useful for assessing bladder cancer risk. PMID- 12544350 TI - Successful diagnosis of orthotopic rat superficial bladder tumor model by ultrathin cystoscopy. AB - PURPOSE: In the orthotopic animal bladder tumor model noninvasive evaluation of the tumor establishment and the therapeutic effect has been difficult. To our knowledge we present the first diagnosis of orthotopic rat superficial bladder tumor model by ultrathin cystoscopy. MATERIALS AND METHODS: The established AY-27 rat bladder carcinoma cell line was transplanted orthotopically into 22 female Fischer F344 rats. A cell suspension containing 4 x 10 AY-27 cells was instilled into the bladder, which had been conditioned with mild acid washing. To evaluate tumor growth serial cystoscopy was performed via the urethra with an ultrathin endoscope (diameter 0.75 mm.) 5 to 14 days after tumor cell inoculation. At intervals after cystoscopic tumor detection the rats were sacrificed for autopsy and histological examination. RESULTS: In all 22 rats the orthotopic bladder tumor was established 7 to 10 days after tumor cell implantation and in most it was superficial. Cystoscopy permitted inspection of the urethra and whole bladder surface. We detected all tumors as broad based papillary mass (minimal lesion 1 mm. or less) and inspected its detailed appearance and accurate location. CONCLUSIONS: The orthotopic rat superficial bladder tumor model and the diagnostic procedure by ultrathin cystoscopy would be ideal system for preclinical evaluation of new potential intravesical therapies. PMID- 12544351 TI - Over expression of metallothionein predicts resistance of transitional cell carcinoma of bladder to intravesical mitomycin therapy. AB - PURPOSE: Metallothionein, a low molecular weight intracellular protein, binds mitomycin with high affinity protecting the tumor DNA. We prospectively studied the relationship of metallothionein expression in bladder transitional cell carcinoma and resistance to intravesical mitomycin. MATERIALS AND METHODS: A series of 45 consecutive patients with superficial transitional cell carcinoma treated with intravesical mitomycin were studied. Resected tumor tissues were stained with metallothionein monoclonal antibody E9. Two pathologists scored staining intensity and distribution. All patients were followed with regular flexible cystoscopy. RESULTS: Median patient age was 73 years (range 44 to 89). Tumor grade was 1 to 3 in 6, 33 and 6 cases, respectively. In 20 patients (44.44%) tumor recurred after mitomycin therapy. Median cytoplasmic staining scores for recurrent and nonrecurrent tumors were 5 (range 0 to 61) and 0 (0 to 14), respectively. Median nuclear staining scores for recurrent and nonrecurrent tumors were 3 (range 0 to 56) and 0 (0 to 11), respectively. Median followup of patients without recurrence was 18 months (range 12 to 36). Nuclear and cytoplasmic staining scores were significantly higher in recurrent than in nonrecurrent tumors. There was no significant relationship of metallothionein expression with tumor grade. CONCLUSIONS: Over expression of metallothionein predicts the resistance of bladder transitional cell carcinoma to intravesical mitomycin therapy. PMID- 12544352 TI - Identification of apoptotic and antiangiogenic activities of terazosin in human prostate cancer and endothelial cells. AB - PURPOSE: It has been suggested that terazosin has an inhibitory effect on prostate tumor growth. We determined if terazosin action contributes to direct suppression of the angiogenic effect. MATERIALS AND METHODS: PC-3 cells and primary cultures of human benign prostatic cells were used in this study. The cytotoxic effect was examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay and lactate dehydrogenase release reaction. The in vivo angiogenic effect was determined in nude mice models, followed by histological examination and quantification by the hemoglobin detection assay. In vitro determination of cell migration, proliferation and tube formation was performed in cultured human umbilical vein endothelial cells. RESULTS Terazosin induced cytotoxicity in PC-3 and human benign prostatic cells with an IC50 of more than 100 microM. The positive terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling and lactate dehydrogenase release reaction was associated with terazosin induced cytotoxicity, indicating apoptotic and necrotic cell death. Furthermore, cytotoxicity due to terazosin action was not a common characteristic of a quinazoline based structure. Terazosin significantly inhibited vascular endothelial growth factor induced angiogenesis in nude mice with an IC50 of 7.9 microM., showing that it had a more potent anti angiogenic than cytotoxic effect. Terazosin also effectively inhibited vascular endothelial growth factor induced proliferation and tube formation in cultured human umbilical vein endothelial cells (IC50 9.9 and 6.8 microM., respectively). CONCLUSIONS: Together our data suggest that terazosin shows direct anti angiogenic activity through the inhibition of proliferation and tube formation in endothelial cells. This action may partly explain the in vivo antitumor potential of terazosin. PMID- 12544353 TI - Enhancement of diethylstilbestrol induced cytotoxicity by bcl-2 antisense oligodeoxynucleotides and a glutathione depletor for prostate cancer. AB - PURPOSE: Bcl-2 has been shown to prolong cancer cell survival by blocking apoptosis and have the function of scavenging reactive oxygen species. We assessed the efficacy of a novel strategy that relies on antisense bcl-2 oligodeoxynucleotides as well as a glutathione depletor combined with diethylstilbestrol (DES) for hormone independent prostate cancer. MATERIALS AND METHODS: The cytotoxic effects of antisense bcl-2 oligodeoxynucleotides and DES on PC-3 cells were determined using spectrophotometry measurement. The expression of Bcl-2 protein was detected by Western blot analysis. Intracellular reactive oxygen species generation was estimated from the amount of intracellular dichlorofluorescein. Apoptosis was determined by the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate-biotin nick end labeling assay. RESULTS: Antisense bcl-2 oligodeoxynucleotides decreased Bcl-2 protein levels and significantly inhibited PC-3 cell growth in a dose dependent manner. Antisense bcl-2 oligodeoxynucleotides significantly enhanced DES induced cytotoxicity. A significant increase in apoptotic cells was induced by the combination of antisense bcl-2 oligodeoxynucleotides and DES compared with the oligodeoxynucleotides alone. The glutathione depletor buthionine sulfoximine significantly decreased the intracellular concentration of glutathione and augmented DES induced cytotoxicity and reactive oxygen species generation in PC-3 cells. Neither the generation of reactive oxygen species nor the intracellular concentration of glutathione was influenced by antisense bcl-2 oligodeoxynucleotide treatment. CONCLUSIONS: Antisense bcl-2 oligodeoxynucleotides significantly enhanced DES induced cytotoxicity in hormone independent prostate cancer cells through the apoptotic pathway independent of augmented reactive oxygen species generation, whereas the glutathione depletor augmented cytotoxicity and reactive oxygen species generation. PMID- 12544354 TI - Response of the fetal sheep bladder to urinary diversion. AB - PURPOSE: In adults urinary diversion results in bladder atrophy and a rapid decrease in contractile function. Little is known about the effects of urinary diversion on bladder development. In this regard we characterized the responses of fetal sheep bladder strips obtained from animals that underwent urinary diversion. MATERIALS AND METHODS: Urinary diversion was performed on fetal sheep after 90 days of gestation (term 147 days) and bladder tissue was obtained 2 weeks later. Contractile and relaxant responses of full-thickness bladder strips were tested. RESULTS: Bladders from fetal sheep subjected to urinary diversion weighed significantly less than control fetal bladders. Histological studies demonstrated marked connective tissue infiltration and the reorganization of smooth muscle elements. Carbachol stimulated a tonic contraction, while field stimulation administered during the tonic contraction elicited a phasic relaxation or a biphasic response, consisting of an initial relaxation and then a phasic contraction in control and diverted bladders. Contractile responses of defunctionalized strips to carbachol were significantly less than those of control bladder strips. Contractile responses of defunctionalized bladder strips to field stimulation at 1 Hz. were significantly greater than those of control strips. Responses of the 2 sets of fetal bladder strips to higher frequencies were similar, as were the contractile responses to adenosine triphosphate and KCl. Field stimulated relaxations in the presence of carbachol stimulated contraction of defunctionalized bladder strips were significantly greater than those of control strips, while the relaxant responses of each set of fetal bladder strips to isoproterenol and nitroprusside were similar. CONCLUSIONS: Urinary diversion in normal fetal sheep resulted in marked structural changes, reduced carbachol stimulation and increased field stimulation relaxation. PMID- 12544355 TI - Endothelin in unilateral ureteral obstruction: vascular and cellular effects. AB - PURPOSE: Unilateral ureteral obstruction results in decreased blood flow and tissue loss in the obstructed kidney. This condition is compensated by increased perfusion and trophic changes in the contralateral kidney. Vascular mediators' effects are central to these changes and of these mediators endothelin is the most potent vasoconstrictor known. We explored the role of endothelin and the effects of endothelin receptor blockade in unilateral ureteral obstruction. MATERIALS AND METHODS: Rats were subjected to unilateral ureteral obstruction for 24 hours. Endothelin-1 mRNA expression was determined in kidney extracts from control, obstructed and contralateral (nonobstructed) kidneys. Cortical and medullary blood flow was determined in control and obstructed kidneys, and after endothelin receptor blockade with bosentan. Apoptotic rates were determined in control and obstructed kidneys after treatment with bosentan using the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end technique. RESULTS After 24 hours of unilateral ureteral obstruction endothelin-1 mRNA expression was increased in the obstructed kidney and decreased in the contralateral kidney. Obstruction was associated with a decrease in renal blood flow, which was reversed by endothelin receptor blockade. Unilateral ureteral obstruction also increased apoptosis, which was blocked by endothelin inhibition. CONCLUSIONS: Endothelin expression increases in the obstructed kidney. Inhibition of its action protects against vascular and cellular changes. Decreased endothelin expression in the contralateral kidney may facilitate trophic changes and compensatory increased blood flow. PMID- 12544356 TI - Evidence of gap junctions in the stable nonobstructed human bladder. AB - PURPOSE: Gap junctions are thought to synchronize muscle cell actions by promoting intercellular communications. Since it is still a matter of debate whether smooth muscle cells in the bladder are electrically coupled, we investigated the occurrence of gap junctions and the gap junction forming protein connexin 45 in normal human detrusor. MATERIALS AND METHODS: Detrusor smooth muscle was obtained during radical cystectomy in 6 patients. For transmission electron microscopy studies samples were high pressure frozen and cryo-fixed. Additional frozen samples were processed for freeze fracture/freeze fracture immunolabeling analyses or for reverse transcriptase-polymerase chain reaction using primers for connexin 45. Furthermore, a set of samples was fixed with Bouin's solution, embedded in paraffin and used for immunohistochemistry with a polyclonal antibody against connexin 45 as well as for in situ hybridization studies with digoxigenin labeled connexin 45 riboprobes. RESULTS: Ultrastructural and freeze fracture studies showed gap junctions at detrusor smooth muscle cells. However, these gap junctions appeared to be small and irregularly shaped. Reverse transcriptase-polymerase chain reaction and in situ hybridization showed that connexin 45 mRNA was expressed in the detrusor muscularis. Furthermore, we detected prominent connexin 45 immunoreactions at detrusor smooth muscle cells. Freeze fracture immunolabeling techniques revealed connexin 45 signals at small gap junctional plaques. CONCLUSIONS: To our knowledge this study is the first to provide morphological, molecular biological and immunohistochemical evidence that smooth muscle cells of stable human detrusor are electrically coupled through gap junctions. These results may contribute to new concepts in the understanding of bladder physiology. PMID- 12544357 TI - Integrins expressed with bladder extracellular matrix after stretch injury in vivo mediate bladder smooth muscle cell growth in vitro. AB - PURPOSE: It is unknown how bladder smooth muscle cells sense extrinsic mechanical stimuli. The integrins are a large versatile family of transmembrane mechanoreceptors that transduce extracellular matrix (ECM) alterations into the cell, thereby, regulating proliferation, differentiation and ECM synthesis. To our knowledge we provide the first evidence that the integrins may be involved in responses to whole bladder distention and bladder smooth muscle cell stretch. MATERIALS AND METHODS: Bladders from 100 to 120 gm. rats were stretched to 40 cm. H2O for 5 minutes. Five to 96 hours after distention whole bladder mRNAs were isolated for analysis of temporal expression of collagen and integrin genes. Separately quiescent primary culture bladder smooth muscle cells from 1-day-old Sprague-Dawley rats were stretched cyclically for 4 hours. Relative expression of select integrin subunit mRNAs was assessed by semiquantitative reverse transcriptase-polymerase chain reaction. Integrin blockade with asparagine glycine-arginine peptides was used to determine the role of integrins in stretch induced proliferation and the cell cycle in bladder smooth muscle cells. RESULTS: Within 24 hours bladder distention stimulated collagen expression 2-fold (type I) and 5-fold (type III). Collagen levels beyond 24 hours were 8-fold (type I) and 2 fold (type III) greater than in controls, revealing an inverse temporal type I-to III ratio beyond 24 hours. Coordinate alterations were observed in integrin and collagen expression. In vitro bladder smooth muscle cell integrin beta1, beta3 and alphav subunit expression was increased by mechanical stretch 2.5, 3.8 and 5 fold, respectively, while alpha1 expression decreased. Asparagine-glycine arginine peptide inhibition of integrin function significantly inhibited stretch induced bladder smooth muscle cell proliferation and exit from the G2/M phase of the cell cycle. CONCLUSIONS: To our knowledge these results demonstrate for the first time that that bladder distention initiates dynamic alterations in ECM expression. The ability of integrin blockade to suppress stretch induced bladder smooth muscle cell proliferation and the coordinate changes in bladder ECM and integrin expression suggest that integrins mediate key responses to mechanical stimuli in the bladder. Furthermore, cell cycle analysis of resting and stretched bladder smooth muscle cells revealed novel avenues for the examination of integrin and stretch regulation of bladder smooth muscle cell growth. PMID- 12544358 TI - Partially purified Grammostola spatulata venom inhibits stretch activated calcium signaling in bladder myocytes and improves bladder compliance in an in vitro rat whole bladder model. AB - PURPOSE: Stretch activated nonselective cationic channels (SACs) are present in urinary bladder myocytes and thought to be activated during bladder filling. We investigated the relationship of stretch induced calcium signaling inhibition in bladder myocytes and bladder compliance modulation in an in vitro whole bladder model. MATERIALS AND METHODS: Grammostola spatulata venom (SpiderPharm, Yarnell, Arizona) was purified by preparative high performance liquid chromatography. The resulting fractions were examined for their ability to inhibit the swelling activated intracellular free Ca2+ signal in cultured bladder myocytes. An in vitro rat whole bladder model was used to examine the effect of venom fractions on compliance, emptying and spontaneous contractions during bladder filling. RESULTS: The gadolinium ion, a SAC inhibitor, and venom fractions caused concentration dependent inhibition of the swelling activated intracellular free Ca2+ signal in bladder myocytes. When tested in a rat isolated whole bladder model, 0.1 and 0.2 mg./ml. partially purified venom produced a significant improvement in compliance (p <0.05), caused significant inhibition of the frequency of spontaneous bladder contractions (mean +/- SEM 35.8% +/- 3.7% and 62.3% +/- 4.4%, respectively, p /=25). Data were obtained from four prospective, double blind, randomized, placebo-controlled clinical trials of the efficacy of reboxetine (8 to 10 mg/d) over 4 to 8 weeks in patients with major depression. In three of the trials, reboxetine produced a significantly greater reduction than placebo in mean HAM-D scores from baseline to the last clinical assessment (p < 0.001). There were significantly more responders to treatment (defined as a reduction in HAM-D score >50% between baseline and the last follow-up observation) treated with reboxetine than placebo in three trials. The overall mean responder rate with reboxetine was 63% (range: 56-74%) compared with 36% (range: 20-52%) with placebo. These results demonstrate that reboxetine is significantly more effective than placebo in a subgroup of patients with severe depression. PMID- 12544376 TI - Anticipatory anxiety in moderately to highly-anxious oral surgery patients as a screening model for anxiolytics: evaluation of alprazolam. AB - Alprazolam, a benzodiazepine anxiolytic, was evaluated in anxious patients prior to oral surgery. This population represents a possible acute screening model for novel anxiolytic agents. Healthy subjects, preselected for a moderate to high degree of dental anxiety based upon Corah's Dental Anxiety Scale, were enrolled in a three-arm parallel design study and randomly assigned to receive double blind placebo (N=15), alprazolam 0.25 mg (N=16) or alprazolam 1 mg (N=16). Subjective self-reported anxiety was rated using the State Anxiety Inventory and visual analog scales. Objective measures included galvanic skin conductance, heart rate variability, blood pressure, pulse rate, and respiration. At 90 minutes after dosing, there were statistically significant (p<0.05) reductions compared with placebo in subjective anxiety and skin conductance mean level for the alprazolam-treated subjects. Changes from pre-dose (mean +/- SEM) at 90 minutes in the placebo, alprazolam 0.25 mg, and alprazolam 1 mg groups were -4.73 +/- 2.79, -13.75 +/- 2.49, and -12.81 +/- 2.32 for the State Anxiety Inventory and 5.44 +/- 6.71, -31.88 +/- 5.88, and -32.34 +/- 5.32 mm for analog anxiety scores. Corresponding skin conductance mean level at 100 minutes in the three groups (respectively) changed 0.64 +/- 0.24, -0.53 +/- 0.21, -0.71 +/- 0.22 microSiemens. The 0.25 mg and 1 mg dosages of alprazolam were not differentiated. Changes in heart rate variability, blood pressure, pulse rate, and respiration did not reflect subjective anxiety. Overall, the oral surgery anticipation anxiety model was found to be a sensitive test for benzodiazepine anxiolytic activity and may represent a potential screening model for evaluation of investigational agents. PMID- 12544377 TI - Psychotropic drugs, cardiac arrhythmia, and sudden death. AB - A variety of drugs targeted towards the central nervous system are associated with cardiac side effects, some of which are linked with reports of arrhythmia and sudden death. Some psychotropic drugs, particularly tricyclic antidepressants (TCAs) and antipsychotic agents, are correlated with iatrogenic prolongation of the QT interval of the electrocardiogram (ECG). In turn, this is associated with the arrhythmia (TdP). This review discusses the association between psychotropic agents, arrhythmia and sudden death and, focusing on TCAs and antipsychotics, considers their range of cellular actions on the heart; potentially pro arrhythmic interactions between psychotropic and other medications are also considered. At the cellular level TCAs, such as imipramine and amitriptyline, and antipsychotics, such as thioridazine, are associated with inhibition of potassium channels encoded by In many cases this cellular action correlates with ECG changes and a risk of TdP. However, not all psychotropic agents that inhibit HERG at the cellular level are associated equally with QT prolongation in patients, and the potential for QT prolongation is not always equally correlated with TdP. Differences in risk between classes of psychotropic drugs, and between individual drugs within a class, may result from additional cellular effects of particular agents, which may influence the consequent effects of inhibition of repolarizing potassium current. PMID- 12544378 TI - Dual monoamine modulation for improved treatment of major depressive disorder. AB - The worldwide scope of depressive illness and lack of fully effective pharmacotherapy mandates significant improvements in treatment paradigms. Current antidepressant medications remain limited by poor efficacy, slow onset of action, and untoward side effects. While the introduction of serotoninspecific reuptake inhibitors (SSRIs) offered significant improvements in tolerability, no improvements in efficacy or speed of onset have been made relative to the traditional and poorly tolerated tricyclic antidepressants (TCA). The dominant efforts toward improving antidepressant medications are guided by cumulative evidence from neurochemical and clinical studies supporting the therapeutic potential of enhancing monoamine function in depression. A number of novel antidepressant drugs, including mirtazapine, milnacipran, venlafaxine, and duloxetine have been developed based on their interaction with both 5-HT and NE. Current clinical evidence suggests that these new agents may offer improved efficacy and/or faster onset of action compared with SSRIs and an improved side effect profile compared with TCAs. Potential neurobiological substrates mediating the enhanced antidepressant activity of dual reuptake inhibitors are discussed. PMID- 12544379 TI - Clozapine pharmacokinetics in children and adolescents with childhood-onset schizophrenia. AB - Clozapine (CLZ) dose-related adverse effects may be more common in children than adults, perhaps reflecting developmental pharmacokinetic (PK) differences. However, no pediatric CLZ PK data are available. Accordingly, we studied CLZ and its metabolites, norclozapine (NOR), and clozapine-N-oxide (NOX) in six youth, ages 9-16 years, with childhood onset schizophrenia (COS). At the time of the PK study, mean CLZ dose was 200 mg (3.4 mg/kg). Serum was collected during week 6 on CLZ before and 0.5-8 h after a morning dose. Serum concentrations were assayed by liquid chromatography/UV-detection. Mean concentration, area-under-the-curve (AUC), and clearance were calculated. CLZ clearance averaged 1.7 L/kg-h. NOR concentrations (410) exceeded CLZ (289) and NOX (63 ng/ml) and AUC(0-8h) of NOR (3,356) > CLZ (2,359) > NOX (559 ng/ml-h) [53, 38, and 9% of total analytes, respectively]. In adults, NOR serum concentrations on average are 10-25% < CLZ, differing significantly from our sample. Dose normalized concentrations of CLZ (mg/kg-d) did not vary with age and were similar to reported adult values. Clinical improvement seen in 5/6 patients correlated with serum CLZ concentrations. In addition, clinical response and total number of side effects correlated with NOR concentrations. NOR (a neuropharmacologically active metabolite) and free CLZ may contribute to the effectiveness and adverse effects in youth. PMID- 12544380 TI - Lithium augmentation of nortriptyline for subjects resistant to multiple antidepressants. AB - Lithium augmentation, the most studied augmentation strategy for depression, has not been evaluated in patients with a history of non-response to multiple antidepressants. The objective of this study was to assess the efficacy of lithium augmentation for patients with a history of treatment resistant depression who also failed a prospective trial of nortriptyline. We enrolled 92 subjects with treatment resistant depression. Treatment resistance was defined by at least one, but no more than five, adequate trials of antidepressants during the current episode. Subjects were treated with nortriptyline (NT) for 6 weeks. Those subjects who tolerated NT for 6 weeks and whose depression did not respond (n=35) were randomized to receive either lithium (n=18) or placebo (N=17) augmentation of nortriptyline for an additional 6 weeks. Response was defined as an equal to or greater than 50% decrease in HAM-D-17 scores. After 6 weeks of double-blind augmentation, 12.5 % of subjects responded to lithium and 20.0% to placebo. Our results revealed no significant difference between lithium and placebo augmentation. While lithium augmentation seems to be useful in depression refractory to a single medication in some studies, our data suggest limited usefulness of this option for patients refractory to multiple treatments. More definitive data await the outcome of the NIMH Sequential Treatment Alternatives to Relieve Depression (STAR*D) study. PMID- 12544381 TI - Influence of DHEA administration on 24-hour cortisol concentrations. AB - DHEA is marketed and readily available as a daily nutritional supplement to counteract the effects of aging. The effect of DHEA administration on 24-hour plasma cortisol profiles has not been investigated. In this single-blind placebo controlled crossover study, the effect of DHEA administration on cortisol concentrations was evaluated in healthy older women and men. Once each morning, subjects took either placebo (Days 1 to 7, and 23 to 29) or oral DHEA 200 mg (Days 8 to 22: doses 1 to 15). Twenty-four hour DHEA and cortisol concentrations were measured on Day 1 (placebo), Day 8 (DHEA dose 1), Day 15 (DHEA dose 8), Day 22 (DHEA dose 15), and Day 29 (placebo washout dose 7). DHEA administration resulted in a decrease in plasma cortisol concentrations (mean, peak, and/or AUC) in healthy older women and men. The cortisol-lowering effect of DHEA was more pronounced in women than in men in our study; pairwise differences in concentrations between days showed that relative to Day 1, cortisol was lower on Days 15, 22, and 29 in women (p = 0.0001) and on Day 15 in men (p = 0.002). The mechanism by which DHEA lowers plasma cortisol concentrations merits further investigation. PMID- 12544382 TI - Usefulness of olanzapine in refractory panic attacks. PMID- 12544383 TI - Olanzapine induced "typical" neuroleptic malignant syndrome. PMID- 12544384 TI - Pancreatitis followed by pericardial effusion in an adolescent treated with clozapine. PMID- 12544385 TI - Efficacy of clozapine in the treatment of atypical antipsychotic refractory schizophrenia: a pilot study. PMID- 12544386 TI - Low-dose risperidone augmentation of antidepressants in nonpsychotic depressive disorders with suicidal ideation. PMID- 12544387 TI - The effect of one-week treatment with venlafaxine on 35% CO2 hyperreactivity in patients with panic disorder: an open study. PMID- 12544388 TI - SSRIs and ejaculation. PMID- 12544390 TI - Immunopathogenesis of chronic allergic conjunctivitis. PMID- 12544391 TI - The role of eosinophils in ocular allergy. PMID- 12544389 TI - Venlafaxine-induced galactorrhea. PMID- 12544392 TI - Cytokines in ocular allergy. PMID- 12544393 TI - Role of chemokines in vernal keratoconjunctivitis. PMID- 12544394 TI - Vernal keratoconjunctivitis. PMID- 12544395 TI - Clinical and immunological features of atopic keratoconjunctivitis. PMID- 12544396 TI - Allergy to topical medications. PMID- 12544397 TI - Diagnostic assays in ocular allergy. PMID- 12544398 TI - Conjunctival provocation testing: overview of recent clinical trials in ocular allergy. PMID- 12544399 TI - New modalities in the treatment of ocular allergy. PMID- 12544401 TI - Genetic susceptibility to infectious diseases. PMID- 12544403 TI - Persistent bacteremia and outcome in late onset infection among infants in a neonatal intensive care unit. AB - BACKGROUND: Persistent bacteremia despite antibiotic therapy has been correlated with adverse outcomes, including focal suppurative complications and death. Coagulase-negative staphylococci (CONS) are the most common cause of nosocomial infection in infants requiring neonatal intensive care and might yield more substantial pathology if infection were persistent. METHODS: To compare the severity and features of persistent infection by CONS with those of other bacteria, we reviewed infants admitted to our neonatal intensive care unit from 1990 through 2001 who developed bacteremia at >5 days of age with recovery of the same bacterial species from blood for >24 h after initiation of antibiotic therapy to which the organism was susceptible. Cases were excluded if a focal complication was already present with the initial positive culture or if the medical record was unavailable. Outcomes of interest included focal suppurative complications, death attributable to infection and duration of hospitalization among survivors. RESULTS: We identified 62 infants with sustained infection, caused by CONS in 30 and by other organisms in 32 [10 Gram-negative, 22 Gram positive (16 Staphylococcus aureus)]. Infants with persistent CONS had significantly lower birth weight and gestational age, but no difference was found for multiple other clinical and demographic risk factors. Indwelling vascular catheters were present at diagnosis in 85% of the infants (CONS 26 of 30, non CONS 27 of 32). Responses of bacteremia to catheter removal vs. in situ treatment did not differ between the groups. No differences were observed for death from all causes (27 vs. 34%), death attributable to infection (6 vs. 12%) or duration of hospitalization among survivors [median (interquartile range): 102 (73 to 167) 107.5 (89 to 130) days]. Focal suppurative complications were significantly more frequent in infants persistently infected with non-CONS (28 vs. 3%; P = 0.01). Duration of persistence correlated with focal complication in non-CONS infants (r = 0.988; P < 0.001). CONCLUSIONS: Although persistent infection with CONS occurs in significantly smaller and less mature infants than with non-CONS, death is no more frequent and focal complications are significantly less frequent. Infants with persistent infection should undergo aggressive evaluation for focal complications, with the yield expected to be higher in those with non-CONS. PMID- 12544402 TI - Impact of the pneumococcal conjugate vaccine on otitis media. AB - CONTEXT: The heptavalent pneumococcal conjugate vaccine (PCV) is recommended for infants to protect against invasive disease, but its impact on otitis might also have public health importance. OBJECTIVE: To examine the impact of PCV on the incidence of otitis media, frequent otitis media and tympanostomy tube procedures and to assess whether the effectiveness of the vaccine wanes after age 24 months and varies by race, sex or season. DESIGN, SETTING AND PATIENTS: From 1995 to 1998, 37 868 children at Kaiser Permanente in Northern California were randomized to receive PCV or a control vaccine in a double blind trial and were followed through April 1999. INTERVENTIONS: Children received a primary series at 2, 4 and 6 months of age and a booster at 12 to 15 months. MAIN OUTCOME MEASURES: Visits for otitis, frequent visits for otitis and tympanostomy tube procedures. Otitis was ascertained from diagnosis checklists routinely marked by physicians. RESULTS: Control children averaged 1.8 otitis visits per year. Children given PCV had fewer otitis visits than control children in every age group, sex, race and season examined. Intention-to-treat analysis permitted rejection of the null hypothesis that PCV is ineffective against otitis media (P < 0.0001). In children who completed the primary series per protocol, PCV reduced otitis visits by 7.8% [95% confidence interval (CI), 5.4 to 10.2%] and antibiotic prescriptions by 5.7% (CI 4.2 to 7.2%). Frequent otitis was reduced by amounts that increased with otitis frequency, from a 10% reduction in the risk of 3 visits to a 26% reduction in the risk of 10 visits within a 6-month period. Tube placements were reduced by 24% (CI 12 to 35%). CONCLUSION: In children followed up to 3.5 years, PCV provided a moderate amount of protection against ear infections while reducing frequent otitis media and tube procedures by greater amounts. PMID- 12544404 TI - Pertussis resurgence in Canada largely caused by a cohort effect. AB - BACKGROUND: Beginning in 1990 Canada experienced a resurgence of pertussis. Changes in incidence and hospitalization according to age in the province of Quebec between 1983 and 1998 were examined to assess the presence of a cohort effect resulting from a poorly protective vaccine. METHODS: The source of data on incident cases was pertussis notifications to the Quebec Ministry of Health and Social Services. Hospitalization data were extracted from the administrative database that collects information on each hospitalization. RESULTS: The mean annual incidence before 1990 was 3.8 cases per 100,000 population which increased to 37.2 thereafter. Infants had the smallest increase (2.7-fold) when compared with children between 1 and 19 years who experienced a 9- to 15-fold increase and with adults (22.5-fold). The mean annual hospitalization rates increased from 2.7 per 100,000 before 1990 to 5.2 afterward. Ninety percent of hospitalizations occurred in children <5 years of age. The proportion of cases in 0- to 4-year-old children decreased, whereas it increased steadily in all other age groups during the entire study period. Between 1990 and 1998 the median age of cases shifted from 4.4 to 7.8 years. Pertussis affected predominantly children who were immunized with a vaccine introduced in the mid-1980s. The evolution of the age distribution of cases paralleled the aging of this cohort with a slow but steady drift of disease from early childhood to adolescence. CONCLUSION: The sudden increase in pertussis incidence in Canada can be largely attributed to a cohort effect resulting from a poorly protective pertussis vaccine used between 1985 and 1998. PMID- 12544405 TI - Effect of yearly vaccinations with live, attenuated, cold-adapted, trivalent, intranasal influenza vaccines on antibody responses in children. AB - BACKGROUND: The cold-adapted, trivalent influenza vaccine (CAIV-T) may become an option for annual vaccination. However, there is little information regarding the immune response to repeated immunization with CAIV-T. OBJECTIVE: To determine the antibody response to repeated immunization with CAIV-T and to compare this with the response after the first CAIV-T immunization. DESIGN AND METHODS: Healthy children were offered CAIV-T immunization for 4 consecutive years, and blood samples were taken from a subset in Years 1, 2 and 4. In Year 4, 156 similarly aged children who had not received influenza vaccine previously were immunized with the same CAIV-T. RESULTS: The H3N2 and B components of the CAIV-T induced high antibody titers in Year 1 that were maintained during 4 years. The H1N1 titers were lower than the H3N2 or B titers. Comparison of the group immunized for 4 consecutive years with the group immunized for the first time revealed the following: (1) before immunization yearly immunized subjects were more likely to be seropositive to each of the three vaccine strains than those immunized for the first time (P < 0.05 for each); (2) after immunization the percentage of seropositive subjects to each of the strains was similar; (3) after immunization titers were higher in the subjects immunized for the first time than those immunized yearly (P < 0.05 for H3N2 and B). CONCLUSION: Yearly vaccination with CAIV-T induced high antibody titers, especially to the H3N2 and B strains in the vaccines. The titers in those immunized with CAIV-T for the first time were higher than in those immunized for 4 consecutive years. PMID- 12544406 TI - Impaired endothelial function in the brachial artery after Kawasaki disease and the effects of intravenous administration of vitamin C. AB - BACKGROUND: Previous studies in patients with a history of Kawasaki disease have focused on vascular endothelial function in coronary arteries, and the endothelial function of systemic arteries is not fully understood. Furthermore the effect of vitamin C on systemic endothelial function after Kawasaki disease has not been elucidated. OBJECTIVES: We attempted to analyze endothelium dependent vasodilatation in the brachial artery after Kawasaki disease by using high resolution ultrasonography and to investigate whether the acute administration of vitamin C could restore such systemic endothelial dysfunction. METHODS: We compared 39 patients (7.1 +/- 2.7 years) 1.0 to 9.6 years after acute Kawasaki disease with 17 matched healthy subjects (7.0 +/- 3.1 years) as controls. Using high resolution vascular ultrasound, we measured brachial artery responses to reactive hyperemia (with increased flow causing endothelium dependent dilatation) and sublingual nitroglycerin (causing endothelium independent dilatation). RESULTS: The percent change in diameter of the brachial artery induced by reactive hyperemia in the patients with a history of Kawasaki disease (6.2 +/- 3.9%) was significantly lower than that in the control group (14.1 +/- 6.8%; P < 0.0001). No significant difference could be found in percent change in diameter induced by sublingual administration of nitroglycerin between the control (33.2 +/- 13.7%) and the patients with a history of Kawasaki disease (30.6 +/- 9.2%; P = 0.49). There was no significant difference in percent change in diameter of the brachial artery induced by reactive hyperemia between the patients who received gamma-globulin (6.0 +/- 4.0%) and those who did not receive gamma-globulin (7.9 +/- 3.3%; P = 0.33). Intravenous infusion of vitamin C significantly increased the percent change in diameter of brachial artery induced by reactive hyperemia in 19 patients with history of Kawasaki disease (6.6 +/- 3.5 to 13.0 +/- 5.5%; P < 0.0001), whereas no significant increase was seen in the percent change in diameter of brachial artery induced by reactive hyperemia in 20 patients with history of Kawasaki disease after placebo administration (6.5 +/- 4.5 to 7.3 +/- 4.9%; P = 0.20). CONCLUSIONS: Our study showed decreased percent change in diameter of the brachial artery induced by reactive hyperemia in patients with history of Kawasaki disease compared with the healthy children, indicating that systemic endothelial dysfunction exits after Kawasaki disease. Although such systemic endothelial dysfunction after Kawasaki disease is not influenced by early treatment with high dose gamma-globulin in the acute stage of Kawasaki disease, it can be restored by the acute intravenous administration of vitamin C. PMID- 12544407 TI - A wider role for congenital cytomegalovirus infection in sensorineural hearing loss. AB - BACKGROUND: Diagnostic problems in identifying congenital infection cases in infancy have thus far impaired the assessment of the role of congenital cytomegalovirus (CMV) infection in the etiology of sensorineural hearing loss (SNHL). OBJECTIVE: To estimate the impact of congenital infection in children with SNHL by detection of CMV DNA in stored samples of neonatal dried blood (dried blood spots test). METHODS: The Guthrie cards of 130 children with hearing loss >40 dB hearing loss were retrieved from the regional screening center. CMV DNA was extracted by thermal shock and amplified by PCR. RESULTS: The percentage of SNHL cases attributable to congenital CMV infection was 10% (9 of 87) in infants whose SNHL had been diagnosed in their first 2 months of life and 34.2% (13 of 38) in children with deafness of unidentified cause that was diagnosed in early childhood. In the latter group 42.7% (12 of 28) of the children with a hearing loss of >70 dB were CMV-positive. CONCLUSIONS: The results suggest that congenital CMV infection has a more relevant role in the etiology of SNHL than previously reported. The data obtained in both groups suggest that 20 to 30% of all deafness cases are caused by CMV. The percent of congenital CMV cases alone appears to account for all the cases previously attributed to all congenital infections. More than 40% of deafness cases with an unknown cause, needing rehabilitation, are caused by congenital CMV. PMID- 12544408 TI - Pathology and causes of death in a series of human immunodeficiency virus positive and -negative pediatric referral hospital admissions in Botswana. AB - BACKGROUND: Little is known about causes of death among children seriously affected by the AIDS epidemic in southern African countries. METHODS: Autopsies were performed on 47 children 1 month to 13 years of age in Francistown, Botswana, between July 1997 and July 1998. RESULTS: Median age was 10 months; 68% were HIV-positive. The leading cause of death was respiratory infection, accounting for 29 of 35 (83%) deaths among HIV-positive and 8 of 12 (67%) deaths among HIV-negative children. Among HIV-positive children, Pneumocystis carinii pneumonia (PCP) was responsible for 31% of all deaths and for 48% of deaths in infants < or =1 year. Among children < or =2 years with cough and dyspnea, age < or =1 year, interstitial infiltrate and HIV positivity were highly predictive of PCP (sensitivity, 100%; specificity, 63%). CONCLUSION: Respiratory disease accounted for most deaths in HIV-positive children. Children < or =1 year who are known or suspected to be HIV-positive and who have cough, dyspnea and pulmonary infiltrates should be treated presumptively for PCP. PMID- 12544409 TI - Pharmacokinetics of nelfinavir and its active metabolite, hydroxy-tert butylamide, in infants perinatally infected with human immunodeficiency virus type 1. AB - BACKGROUND: In children younger than 2 years of age vertically infected with HIV 1, the recommended pediatric dosing regimen for nelfinavir (20 to 30 mg/kg three times a day) provides insufficient drug exposure. This study was conducted to determine the steady state pharmacokinetics of nelfinavir and its active metabolite, M8, in this population. METHODS: Fourteen infants (2.3 to 8.5 months) underwent 18 intensive pharmacokinetic studies of nelfinavir and M8 at steady state. Nelfinavir and M8 concentrations were measured by high performance liquid chromatography coupled with mass spectrometry, and individual pharmacokinetic values were determined. RESULTS: A mean nelfinavir daily dose of 135.7 +/- 18.8 mg/kg (twice or three times a day) resulted in median C(min), C(max), area under the plasma concentration-time curve (AUC(0-24 h)) and CL/ for nelfinavir of 0.627 mg/l, 2.39 mg/l, 30.6 mg*h/l and 4.2 liters/h/kg, respectively. When normalized for a daily dose of nelfinavir of 150 mg/kg/day, 16.7% of C(max) and 27.8% of AUC(0-24 h) values were below the tenth percentile for adult values. CONCLUSIONS: During the first year of life, nelfinavir requirement is much higher than in older children and adults to obtain similar drug exposure. The mechanisms underlying such differences may involve higher first past metabolism and/or drug interactions or might be related to feeding conditions. PMID- 12544410 TI - Adherence to prescribed antiretroviral therapy in human immunodeficiency virus infected children in the PENTA 5 trial. AB - BACKGROUND: Most studies of adherence to highly active antiretroviral therapy in children have been retrospective or cross-sectional. Factors relating to the caregiver, the child and the medication are all considered to be important for good adherence. METHODS: Adherence with taking prescribed medication was assessed by questionnaires completed at 4, 12, 24 and 48 weeks by caregivers of previously untreated HIV-infected children participating in the PENTA 5 trial, which was designed to evaluate different dual nucleoside reverse transcriptase inhibitor therapy combinations with and without the protease inhibitor nelfinavir. The effects of several factors on adherence and the effect of adherence on virologic suppression were assessed by multivariate logistic regression. RESULTS: Caregivers returned 266 questionnaires including at least 1 for 108 (84%) children in the trial. Nelfinavir was reported to be the most difficult drug to take (38% of questionnaires), but the difficulty decreased over time, P = 0.02. Comments on difficulties in taking and remembering drugs related to fear of disclosure and to unpleasant characteristics of the drugs. Full adherence was reported in 74% of questionnaires, did not change over time and was reported more frequently in children older than 10 years and those with symptomatic HIV disease. More children reporting full adherence achieved HIV RNA <400 copies/ml (e.g. at 48 weeks 79% vs. 50% reporting some nonadherence; overall P = 0.01). CONCLUSION: Good adherence with taking prescribed medication was associated with virologic response. Social factors were important in explaining nonadherence. PMID- 12544411 TI - Age-related immune reconstitution during highly active antiretroviral therapy in human immunodeficiency virus type 1-infected children. AB - OBJECTIVES: To evaluate the immune reconstitution in HIV-1-infected children in whom highly active antiretroviral therapy (HAART) controlled viral replication and to assess the existence of a relation between the magnitude of this restoration and age. METHODS: All HIV-1-infected children in whom a new HAART decreased plasma viral load below 400 copies/ml after 3 months of therapy were prospectively enrolled in a study of their immune reconstitution. Viral load, lymphocyte phenotyping, determination of CD4+ and CD8+ T cell receptor repertoires and proliferative responses to mitogens and recall antigens were assessed every 3 months during 1 year. RESULTS: Nineteen children were evaluated. Naive and memory CD4+ percentages were already significantly increased after 3 months of HAART. In contrast to memory CD4+ percentages, naive CD4+ percentages continued to rise until 12 months. Age at baseline was inversely correlated with the magnitude of the rise in naive CD4+ cells after 3, 6 and 9 months of therapy but not after 12 months. Although memory and activated CD8+ cells were already decreasing after 3 months, abnormalities of the CD8 T cell receptor repertoire and activation of CD8+ cells persisted at 1 year. HAART increased the response to mitogens as early as 3 months after starting therapy. CONCLUSIONS: In children the recovery of naive CD4+ cells occurs more rapidly if treatment is started at a younger age, but after 1 year of viral replication control, patients of all ages have achieved the same level of restoration. Markers of chronic activation in CD8+ cells persist after 1 year of HAART. PMID- 12544413 TI - Metabolic complications of antiretroviral therapy in children. AB - Survival in HIV-infected children has greatly improved with the introduction of highly active antiretroviral therapy. Children are more vulnerable than adults to metabolic side effects of therapy because of its potential impact on growth and the children's likely greater cumulative exposure. This review summarizes the epidemiology and management of lipodystrophy, dyslipidemia, insulin resistance, hyperlactatemia, osteopenia and growth failure in HIV-infected children. PMID- 12544412 TI - Endocrinologic and immunologic factors associated with recovery of growth in children with human immunodeficiency virus type 1 infection treated with protease inhibitors. AB - INTRODUCTION: Growth failure is a common presenting sign in children with HIV disease and is a sensitive indicator of disease progression in children with AIDS. Highly active antiretroviral therapy (HAART) is associated with a significant decrease in viral load and a subsequent rise in CD4+ T cell counts in HIV-1-infected children and also with increased height and weight. The underlying mechanisms of catch-up growth during HAART are yet unknown. METHODS: Height and weight measurements, blood sample analyses for HIV-1 RNA and peripheral blood CD4+ T cell counts were obtained twice within 1 month before the start of HAART and after 12, 24, 36 and 48 weeks of treatment. Serum concentrations of insulin like growth factor I (IGF-1), IGFs complexed to specific, structurally homologous binding proteins (IGFBP-3), cortisol, free thyroxine and tumor necrosis factor alpha (TNF-alpha) were measured before the start of therapy and after 24 weeks. In addition serum IGF-1 and IGFBP-3 values were determined after 48 weeks. RESULTS: Twenty-seven HIV-1-infected children with a median age of 5.5 years (range, 0.3 to 14.9 years) were included. Overall no significant changes in height and body mass index (BMI) z scores were observed. The median baseline plasma viral load of 68,800 copies/ml decreased to less than the detection limit of 500 copies/ml in 80% of the children after 48 weeks. TNF-alpha values were elevated (44 pg/ml) at baseline and decreased significantly to 37 pg/ml after 24 weeks. At baseline elevated TNF-alpha was observed in 78%, which decreased to 55% after 24 weeks. Baseline free thyroxine and cortisol values were normal and did not change during therapy. Baseline serum of IGF-1 and IGFBP-3 concentrations were normal, but IGF-1 tended to be lower than IGFBP-3. Both values increased significantly after the initiation of therapy. IGFBP-3 decreased after 48 weeks whereas IGF-1 stabilized. The increase in IGF-1 was significantly higher in children in whom the BMI and length (after correction for age and sex) increased the most. CONCLUSION: Hypothyroidism and adrenal axis abnormalities are not associated with restoration of growth after the initiation of antiretroviral therapy in HIV-1-infected children. The combination of relatively high serum IGFBP-3 concentration and relatively lower serum IGF-1 suggests the presence of a growth hormone-resistant state. During treatment with a protease inhibitor containing regimen, decreased serum IGFBP-3 and stabilization of IGF-1 after a significant initial increase suggest restoration of normal sensitivity to growth hormone and recovery to an anabolic condition. PMID- 12544414 TI - Case-control studies. PMID- 12544415 TI - Update on phagocytic defects. PMID- 12544416 TI - Anti-inflammatory effects of local anesthetic agents. PMID- 12544417 TI - Effects of volatile and intravenous anesthetic agents on neutrophil function. PMID- 12544418 TI - Implications of anesthesia for infection and wound healing. PMID- 12544419 TI - Apoptosis in critical illness. PMID- 12544420 TI - Implications of the cytosolic phospholipase A2 knockout mouse on perioperative therapeutics. PMID- 12544421 TI - Inhaled nitric oxide in acute respiratory distress syndrome: from bench to bedside. PMID- 12544423 TI - New year, new challenges. PMID- 12544424 TI - Cross-linking of glycated collagen in the pathogenesis of arterial and myocardial stiffening of aging and diabetes. AB - The normal aging process is often accompanied by arterial wall stiffening and by a decrease in myocardial compliance. These processes contribute to isolated systolic hypertension and diastolic heart failure, which lead to substantial morbidity and mortality among older individuals. Patients with diabetes manifest arterial stiffening and diastolic dysfunction at a younger age. This leads to the concept that the mechanism that underlies changes in vascular mechanical properties during aging is accelerated in diabetes. The Maillard reaction or advanced glycation of proteins occurs slowly in vivo with normal aging and at an accelerated rate in diabetes. Advanced glycation end-products (AGEs) that form during the Maillard reaction are implicated in the complications of aging and diabetes. The formation of AGEs on vascular wall and myocardial collagen causes cross-linking of collagen molecules to each other. This leads to the loss of collagen elasticity, and subsequently a reduction in arterial and myocardial compliance. Aminoguanidine, an inhibitor of AGE formation, is effective in slowing or preventing arterial stiffening and myocardial diastolic dysfunction in aging and diabetic animals. In aged and diabetic animals, agents that can chemically break pre-existing cross-linking of collagen molecules are capable of reverting indices of vascular and myocardial compliance to levels seen in younger or non-diabetic animals. These studies suggest that collagen cross-linking is a major mechanism that governs aging and diabetes-associated loss of vascular and cardiac compliance. The development of AGEs cross-link breakers may have important role for future therapy of isolated systolic hypertension and diastolic heart failure in these conditions. PMID- 12544425 TI - What defines an adolescent as having hypertension? PMID- 12544426 TI - X-linked quantitative trait locus for susceptibility to renal injury. PMID- 12544427 TI - Genetic influences in human hypertension. PMID- 12544428 TI - The world of endothelin in the brain of the stroke-prone spontaneously hypertensive rat. PMID- 12544429 TI - Endothelin-dependent modulation of vasoconstrictor action in the coronary circulation in early salt-induced experimental hypertension. PMID- 12544430 TI - Omapatrilat: still a promise in salt-sensitive hypertension? PMID- 12544431 TI - Sodium intake and vessel wall properties of large arteries. PMID- 12544432 TI - Renal dopamine receptors and hypertension. PMID- 12544433 TI - Nephrin: a pivotal molecule in proteinuria influenced by angiotensin II. PMID- 12544434 TI - Blood pressure distribution in a Hungarian adolescent population: comparison with normal values in the USA. AB - OBJECTIVES: To describe the blood pressure characteristics and distribution of all adolescent high school students (aged 15-18 years) in Debrecen (total population 230 000), Hungary. To define threshold values for normal blood pressure by age-, sex- and height-specific groups. To compare our data with results from a USA meta-analysis, which forms the basis of current guidelines. PARTICIPANTS AND METHODS: All young people attending high school in Debrecen (final sample = 10 359) participated in the study. After they had rested for 10 min, three blood pressure measurements were taken from the right upper arm, separated by 5 min intervals. All measurements were obtained by a validated, automated, digital Omron M4 device. RESULTS: The 50th, 90th and 95th percentile values of blood pressure were defined by dividing the adolescent population into age-, sex- and height-specific subgroups. In comparison with USA guidelines, in our sample the systolic blood pressure of boys in the different subgroups was 6 11 mmHg greater, whereas this difference was less marked for girls (1-5 mmHg). There were no marked differences in diastolic blood pressure, but our values were slightly lower. CONCLUSIONS: Our findings demonstrate the influence of geographical and ethnic variations on blood pressure. Acceptance and use of non population-specific blood pressure distributions may lead to under- or overdiagnosis of adolescent hypertension. The use of geographically more relevant data should be encouraged. PMID- 12544435 TI - Removing confounders from the relationship between mortality risk and systolic blood pressure at low and moderately increased systolic blood pressure. AB - OBJECTIVE: To assess the relationship between mortality risk and systolic blood pressure (SBP) at low and moderately increased SBP (less than the 70th percentile) before and after correcting for the regression-dilution bias and J curve effects. DESIGN: Cohort study. SETTING: The First National Health and Nutrition Examination Survey Epidemiologic Follow-Up Study. PARTICIPANTS: The 6839 individuals who participated in the 1982-1984 survey for whom there were no missing data (age range 34-87 years). MAIN OUTCOME MEASURE: Cardiovascular disease mortality (n = 678) during a 9-year follow-up. METHODS: Corrections were made for the regression-dilution bias by using average SBP during the decade before baseline as the mortality predictor, and for J-curve effects by excluding individuals who exhibited high age-stratified mortality rates and a decrease in SBP. Cox's regression was used to analyse the follow-up relationship between mortality risk and SBP. RESULTS: The corrected relative cardiovascular disease mortality risk was 1.23 (95% confidence interval (CI), 1.16 to 1.31) for a 10 mmHg increase in SBP. The relationship was monotonically positive starting at the lowest SBP category in the analysis (< 115 mmHg), and robustly so above about the 32nd percentile (120 mmHg). The equivalent uncorrected result was 1.08 (95% CI, 1.05 to 1.13), and the relationship was J-shaped and became positive above about the 68th percentile (135 mmHg). Below the 70th percentile of SBP, individuals in the corrected analysis were 89% of all individuals. CONCLUSION: These two corrections transformed the relationship between mortality risk and SBP at low and moderately increased SBP from no association to a robustly positive association starting at 120 mmHg, for the majority of individuals. PMID- 12544436 TI - Hypertension and changes of cognitive function in 81-year-old men: a 13-year follow-up of the population study "Men born in 1914", Sweden. AB - BACKGROUND: The relationship between blood pressure levels and decline of cognition has been discussed previously, but little is known about the confounding effect of lifestyle factors, antihypertensive treatment and disease in the elderly. OBJECTIVE: To examine the association between systolic (SBP) and diastolic (DBP) blood pressure, changes of DBP and SBP and hypertension (HT) on cognitive function, while controlling for confounding factors. DESIGN: A longitudinal cohort study of men born in 1914 and residing in the municipality of Malmo studied at age 68 and 81 years. SETTING: Probands invited to a university clinic. SUBJECTS: One hundred and eighty-six out of 281 invited men still alive in 1995/6 participated in the most recent follow-up. MAIN OUTCOME: Change of the cognitive performance in the verbal (Paired Associates and Synonyms), spatial (Block Design and Benton Visual Retention test) and speed (Digit Symbol Substitution) functions. RESULTS: DBP by tertiles at 68 years, but not HT, was inversely related to verbal, spatial and speed performance at 81 years. Only spatial function was related to SBP at 68 years. The association between DBP and SBP by tertiles, and spatial functions (Block Design and Benton Visual Retention test) remained after controlling for education, marital status, smoking, alcohol and physical activity, and intermediates such as arteriosclerotic manifestations (block design, beta = 0.17; = 0.029) in multiple regression models. A decrease in DBP was likewise related to spatial (beta = 0.16; = 0.049) and speed performance (beta = 0.17; = 0.039) in the same regression model. CONCLUSIONS: Evidence is found to support the hypothesis that hypertension, especially high DBP in late midlife, is associated with a decline in spatial performance of cognitive functions in elderly men. PMID- 12544437 TI - X-linked locus associated with hypertensive renal disease susceptibility in Dahl rats. AB - OBJECTIVE: There is increasing evidence that genetic factors contribute to renal disease susceptibility associated with essential hypertension. To what extent these genetic factors act independently of hypertension susceptibility remains undetermined. The present study was undertaken to assess the potential chromosome X influence on target organ renal disease in the Dahl rat model of salt-sensitive hypertension. SUBJECTS AND METHODS: Dahl S, Dahl R, F1(RXS), F1(SXR) and F2(RXS) rat male populations were phenotyped for hypertensive renal disease by measuring the percent of incidence of the Grade IV Raij renal pathology score. Six chromosome X markers informative for our (RXS) intercross were analyzed in our F2 rat population (n = 105) for co-segregation with hypertensive renal disease and blood pressure characterized by radiotelemetry. RESULTS: Comparison of the incidence of renal disease (histologically determined) between F1 reciprocal intercross male progenies reveals a significant chromosome X effect on renal disease [percent incidence of Grade IV Raij renal pathology score in F1 (R female S male) male rats = 2.75 +/- 0.66, and in F1 (S female R male) male rats = 0.67 +/- 0.42; = 0.02]. QTL analysis on an F2(RXS) male population phenotyped for renal disease susceptibility (percent incidence of Grade IV Raij renal pathology score) detects significant linkage to DXRat98 (likelihood ratio statistic = 9.4, P = 0.00223) on chromosome X, corroborating X-linkage of renal disease susceptibility in Dahl rats. CONCLUSIONS: Our results demonstrate the existence of an X-linked locus associated with hypertensive renal disease susceptibility in Dahl rats. Furthermore, the chromosome X markers tested did not co-segregate with hypertension, indicating that the gene(s) on chromosome X influence renal disease susceptibility independent of blood pressure. PMID- 12544438 TI - Major gene effects in systolic and diastolic blood pressure in families receiving a health examination in Taiwan. AB - BACKGROUND: Genetic factors play an important role in determining blood pressure values. Strong familial aggregation and estimated heritability values around 0.4 0.6 were reported in a white population. OBJECTIVES: To investigate the genetic components and mode of inheritance of blood pressure in Taiwan. DESIGN: A cross sectional family study based on an adult population undergoing a health examination. SETTING AND PARTICIPANTS: A total of 1313 adults and their spouses or first-degree relatives, or both, underwent a health examination in a tertiary university hospital from August 1998 to September 1999. MAIN OUTCOME MEASURES: Genetic analyses, including comingling analysis, familial correlation and complex segregation analysis, were used to detect the genetic components and the mode of inheritance of blood pressures. RESULTS: In both systolic and diastolic blood pressure, the comingling analyses demonstrated that a more than one-component distribution provided the best fit for the data. The familial correlation coefficients showed significant parent-offspring and sibling correlation. Complex segregation analyses showed major gene effects in controlling systolic and diastolic blood pressure. CONCLUSIONS: There were significant genetic components in blood pressure among Taiwanese. Further investigation of genomic loci for the control of blood pressure is indicated. PMID- 12544439 TI - Effects of ACE I/D and AT1R-A1166C polymorphisms on blood pressure in a healthy normotensive primary care population: first results of the Hippocates study. AB - BACKGROUND: Several studies have assessed the relationship between the angiotensin-converting enzyme (ACE) I/D or angiotensin II type 1 receptor (AT(1)R)-A C polymorphisms and blood pressure (BP). Since most data have been obtained in selected populations, the present study was performed in a healthy normotensive primary care population. OBJECTIVE: To investigate the individual effects of the aforementioned polymorphisms and their interaction on BP. METHODS: This cross-sectional study included 198 healthy subjects. Office BP was measured and polymorphisms were genotyped (polymerase chain reaction). Polymorphism interaction was tested using the following model: systolic blood pressure (SBP) (or diastolic blood pressure, DBP) = b(0)+ b(1)X + b(2)Y + b(3)XY, in which X and Y represent the polymorphisms' risk alleles. RESULTS: The ACE I/D polymorphism was associated with SBP (P = 0.002) and DBP (P = 0.004); highest pressures tracked with the DD genotype. Furthermore, in multiple linear regression analysis the ACE D allele was associated with SBP (P = 0.005) and DBP (P = 0.001), when adjusted for body mass index (BMI) and age. With respect to the AT(1)R-A C polymorphism, SBP was highest in the CC genotype (P = 0.025). In linear regression analysis the C allele was not associated with SBP. No synergistic effect of ACE D and AT(1)R C alleles on BP was found. Nevertheless, highest DBP tracked with the DDCC combination in comparison with other homozygous allele combinations (P = 0.030). CONCLUSIONS: This study confirmed an association of ACE I/D and AT(1)R-A C polymorphisms with BP in a healthy normotensive primary care population. Although synergistic effect of both polymorphisms on BP does not seem to be present, an additive effect on DBP is likely. PMID- 12544440 TI - Contrasting associations between aldosterone synthase gene polymorphisms and essential hypertension in blacks and in whites. AB - BACKGROUND: Genetic variability in the gene for aldosterone synthase--a key enzyme in the production of aldosterone--can affect sodium homeostasis and thereby blood pressure. The possibility of impaired aldosterone production for the development of hypertension is of particular relevance in black Afro Caribbeans exposed to a high dietary sodium intake. OBJECTIVES: To compare the frequency of three variants (-344C/T, intron 2 conversion, and the K173R polymorphism) of the aldosterone synthase gene in blacks and whites, and to determine any association of the variants with hypertension. DESIGN AND METHODS: We made case-control comparisons of the three gene variants in relation to ethnic background and to essential hypertension in 193 white (51% hypertensive) and 245 black individuals (59% hypertensive) living in south London. RESULTS: The frequency of each of the variants was significantly different between the two ethnic groups. The T and the K alleles were more frequent in the black participants (79 compared with 50% for the -344T allele and 81 compared with 50% for K173 allele), whereas the frequency of the intron 2 conversion allele was much lower in that group (8 compared with 38%). None of these variants was associated with essential hypertension in the black participants. In contrast, in the white participants there was a significant and graded association between the intron 2 conversion allele and essential hypertension (odds ratio 1.86, 95% confidence interval 1.16 to 2.98; = 0.01). Moreover, among the white population, the presence of homozygosity both of the T allele and of the intron 2 conversion alleles was associated with a much greater frequency of hypertension (71 compared with 43%; chi(2) = 0.03). CONCLUSIONS: The contrasting associations between these variants and essential hypertension do not necessarily exclude the possibility that other, as yet undefined, variants of the aldosterone synthase gene could be linked with hypertension in black people. Nonetheless, the strong association between the intron 2 conversion allele and essential hypertension in the white population reinforces the view that the increased blood pressure may be due, at least in part, to abnormal expression of enzymes involved in the metabolism of adrenal mineralocorticoids. PMID- 12544441 TI - Effects of alcohol intake on endothelial function in men: a randomized controlled trial. AB - BACKGROUND: Regular light consumption of alcohol appears to reduce the risk of cardiovascular disease, whereas in heavier drinkers the opposite effect is seen. This biphasic relationship could partly be due to contrasting actions of low and high alcohol intake on endothelial function. OBJECTIVE: To determine whether reducing alcohol intake in moderate-to-heavy drinkers (40-110 g/day) would improve conduit artery endothelial function as assessed by post-ischaemic brachial artery flow-mediated dilatation (FMD). METHODS: In a two-way cross-over study, 16 healthy men either substituted their usual alcohol intake with a 0.9% alcohol beer or maintained their usual alcohol intake during sequential 4-week periods. At the end of each period of FMD and glyceryl trinitrate-induced brachial artery dilatation, blood pressure, plasma lipids, homocysteine and biomarkers of alcohol consumption (gamma-glutamyl transpeptidase) and endothelial function (E-selectin, von Willebrand factor, endothelin-1) were assessed. RESULTS: The participants reduced their alcohol intake from 72.4 to 7.9 g/day. This self-reported reduction in alcohol intake was corroborated by significant decreases in gamma-glutamyl transpeptidase (24%). The decrease in alcohol intake resulted in reductions in total cholesterol (5%), high-density lipoprotein cholesterol (17%), homocysteine (9%) and systolic and diastolic blood pressure [5 mmHg (P = 0.01) and 4 mmHg (P = 0.003), respectively]. There was no effect of alcohol on FMD (6.23 +/- 0.75% compared with 6.24 +/- 0.71%, P = NS), glyceryl trinitrate-induced vasodilatation, E-selectin, endothelin-1 and von Willebrand factor. CONCLUSION: Substantial reduction in alcohol intake in healthy moderate to-heavy drinkers does not improve endothelial function as measured by post ischaemic flow-mediated dilatation of the brachial artery or biomarkers of endothelial function. PMID- 12544442 TI - Altered regulation of endothelin A receptor subtype in the cerebral arterioles in response to a Japanese-style diet, in stroke-prone hypertensive rats. AB - OBJECTIVE: To investigate the expression of endothelin (ET)-1 and its receptors in the cerebral arterioles of stroke-prone (spSHR) and control spontaneously hypertensive rats (SHRs) and the changes in endothelin receptor subtypes A and B density elicited by a stroke-permissive diet, before the development of stroke. METHODS: Six-week-old SHRs (n=11) and spSHRs (n=11) were assigned to either a regular or a "Japanese"-style diet, in addition to 1% NaCl in the drinking water, for 4 weeks. Cryosections (10 microm thick) of rat brain were assessed for endothelin receptor distribution and density by autoradiography with [125I]ET-1 (10(-10) mol/l) in the presence of cold ET-1 (10(-6) mol/l) or the peptide antagonists BQ-123 (10(-6) mol/l) or BQ-788 (10(-6) mol/l). Reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry were used to detect specific mRNAs and localize immunoreactive ET-1 and ET(A) and ET(B). RESULTS: In both strains, immunoreactive ET-1 was detected in the endothelium of cerebral arterioles, and RT-PCR and autoradiography demonstrated the coexistence of both receptor subtypes in brain homogenates and the cerebral arteriole walls, respectively. With the regular diet, the ET(A) receptor density was lower in SHRs than in spSHRs (P = 0.007), whereas the ET(B) receptor density was similar (P = NS). The Japanese-style diet increased the density of ET(A) receptors (P = 0.006) in SHRs, but decreased it (P = 0.019) in spSHRs. No effect was seen on ET(B) receptor density. CONCLUSIONS: ET(A) and ET(B) receptor subtypes are expressed in the wall of cerebral arterioles of SHRs and spSHRs. The latter strain showed a marked increase in ET(A) receptor density under a regular diet, in addition to an altered regulation in response to a stroke-permissive diet. PMID- 12544443 TI - Contribution of endogenous endothelin in the enhanced coronary constriction in DOCA-salt hypertensive rats. AB - OBJECTIVE: The present study was performed to evaluate the hypersensitivity to vasoconstrictors in coronaries from uninephrectomized hypertensive rats (HTR), after a 2-week deoxycorticosterone acetate (DOCA)-salt treatment, in comparison with uninephrectomized age-matched normotensive rats (NTR). DESIGN AND METHODS: Coronary resistance was recorded from isolated Langendorff hearts perfused at a constant flow rate. RESULTS: Cumulative dose-response curves to vasopressin, angiotensin II and endothelin in HTR showed an enhanced maximal response, in comparison with NTR (P< 0.05). In contrast, the sensitivity to U-46619, a thromboxane-mimetic agonist, was reduced in HTR in comparison with NTR (P< 0.05). In the presence of ET(A)/ET(B)-receptor antagonists, LU-302 872 (10 micromol/l) and PD-142 893 (0.1-1 micromol/l), cumulative dose-response curves to vasopressin and angiotensin II showed a reduced maximal response in HTR compared with NTR (P< 0.05). LU-302 872 did not change the responsiveness to U-46619 in both groups. Perfusion of hearts from NTR with a subpressor concentration of endothelin-1 (10 pmol/l) potentiated the responsiveness to vasopressin and angiotensin II, but not that of U-46619 (P< 0.05). Hypertension did not alter the dose-response curves obtained with phorbol 12-myristate 13-acetate, an activator of protein kinase C, Bay K 8644, a L-type calcium-channel activator, and KCl. Measurement of endothelin release by radioimmunoassay in the coronary effluent, before and during dose-response curves to vasopressin, angiotensin II and U-46619, showed no significant increase by the vasoconstrictors, although basal endogenous endothelin was increased in HTR (P< 0.05). CONCLUSION: Two-week DOCA-salt hypertension is associated with enhanced coronary vasoconstrictor effects of endothelin, vasopressin and angiotensin II. An increased basal release of endogenous endothelin in coronaries from HTR, along with an enhanced responsiveness of the coronary smooth muscle to endothelin, may contribute to the potentiated response to vasoconstrictors. L-type calcium-channels and protein kinase C are not involved in this increased coronary reactivity to vasoconstrictors in HTR. PMID- 12544444 TI - Comparison of the cardiovascular protection by omapatrilat and lisinopril treatments in DOCA-salt hypertension. AB - OBJECTIVE: To compare the cardiovascular protection provided by omapatrilat and lisinopril in an experimental model of hypertension. METHODS: Four-week deoxycorticosterone acetate (DOCA)-salt hypertensive (HT) and age-matched normotensive (NT) rats were treated either with omapatrilat (40 mg/kg per day) or lisinopril (20 mg/kg per day) for 2 weeks before sacrifice, and compared with untreated HT and NT rats sacrificed at ages corresponding to either before or after the drug regimens. RESULTS: Systolic arterial pressure (SAP) of 2 and 4 week HT rats was increased in comparison to age-matched NT rats (P <0.05). Treatment with omapatrilat or lisinopril reduced SAP in HT (P <0.05) similarly by about 10%. Cardiac interstitial collagen, perivascular collagen and media/lumen ratio of coronary arterioles were increased in HT rats. Both treatments partially prevented the rise in perivascular collagen and completely corrected the increased media/lumen ratio in small arterioles from HT (P <0.05). In contrast to NT rats, only a weak coronary dilatation to bradykinin was observed in Langendorff hearts isolated from untreated-HT. This response was slightly improved by lisinopril and markedly improved by omapatrilat (P <0.05). The coronary dilatation to SNP which was reduced in 4-week HT (P <0.05), was partially improved by omapatrilat treatment but not by lisinopril. The enhanced superoxide anion production in aorta from HT rats was partially corrected with omapatrilat and lisinopril. Finally, omapatrilat, unlike lisinopril, markedly reduced mortality in a more severe form of DOCA-salt hypertension. CONCLUSIONS: Omapatrilat and lisinopril regressed coronary remodelling and cardiac collagen deposition, and reduced vascular oxidative stress in DOCA-salt hypertensive rats. However, despite similar antihypertensive efficacy, omapatrilat was superior to lisinopril in improving the endothelial-dependent coronary dilatation, suggesting a better vascular protection in the DOCA-salt model of hypertension. PMID- 12544445 TI - Altered structure and distensibility of arteries in salt-fed rats. AB - BACKGROUND AND OBJECTIVES: Habitual high-sodium diet may cause stiffening of arteries. The aim of this study was to investigate the long-term effects of physiologically relevant high-sodium diet on the structure and distensibility of arteries in rats. METHODS: Adult male Sprague-Dawley rats were fed 2% NaCl diet for 3 or 6 months; rats fed 0.7% NaCl diet were controls. Pressure-volume (distensibility) relationships were measured in the presence and absence of calcium in excised, in-vitro perfused segments of right carotid artery and of second order mesenteric arteries. The left carotid artery and the remaining mesenteric arteries of rats were perfused in situ with papaverine followed by fixative at 100 mmHg, and then embedded in epoxy for morphometric measurements. RESULTS: The tail systolic blood pressure (SBP), and in subgroups of rats, the directly measured mean arterial pressure (MAP), of salt-fed rats at 3 and 6 months were unchanged. At 3 months, there was dilatation (increased lumen area) of both carotid and mesenteric arteries of salt-fed rats, without a change in distensibility. At 6 months, the lumen area of carotid arteries of salt-fed rats returned to control value (inward remodeling), and carotid artery distensibility remained unchanged. At 6 months, there was further dilatation (P <0.01) and reduced distensibility (P =0.01) of mesenteric arteries in salt-fed rats. CONCLUSIONS: A three-fold increase in dietary sodium intake leads to dilatation of arteries in normotensive rats. When there is compensatory remodeling, the distensibility of arteries remains unchanged; when compensation is lacking, unopposed dilatation is associated with reduced distensibility. PMID- 12544446 TI - Oxidative stress in juvenile essential hypertension. AB - OBJECTIVE: Oxidative stress, an antioxidant/pro-oxidant imbalance, in patients with juvenile essential hypertension was measured via several biochemical parameters. As the blood pressure is associated with the body mass index (BMI), results were compared with those on BMI-matched controls. DESIGN AND SETTING: A prospective observational study at a university teaching hospital. PATIENTS: Children and adolescents with essential hypertension (mean standard deviation: age 14.4 +/- 3.1 years, BMI 25.0 +/- 6.9 kg/m(2), n = 52) before any treatment, and controls with a similar BMI distribution (age 14.3 +/- 4.3 years, BMI 24.4 +/ 6.6 kg/m(2), n = 48). METHODS: Measurements were made of the plasma levels of (1) nitrites + nitrates, an indirect measure of available nitric oxide; (2) lipid peroxidation end-products, as malondialdehydes and free thiols; and (3) the redox status of the red blood cell glutathione, as a new oxidative stress parameter. RESULTS: There were decreased plasma levels of nitrates and increased levels of lipid peroxidation end-products in the hypertensive patients, resulting in a consistent increase in the plasma lipid peroxidation/nitric oxide ratio as compared with the controls with the same BMI (P <0.01). This ratio additionally correlated directly with both the systolic and diastolic blood pressures for the overall patient population (P <0.001). A significant glutathione depletion in the red blood cells resulted in an elevated ratio of oxidized/reduced forms with a reduced antioxidant protective capacity in the hypertensive patients versus the BMI-matched controls (P <0.001). CONCLUSIONS: The presence of systemic oxidative stress was proven in hypertensive children and adolescents, irrespective of their BMI. PMID- 12544448 TI - Central cardiovascular action of urotensin II in conscious rats. AB - OBJECTIVE: To examine the central cardiovascular action of urotensin II in conscious rats. METHODS: Intracerebroventricular (ICV) injections of urotensin II (1 and 10 nmol) were carried out in conscious Wistar rats. The effects of intravenous (i.v.) urotensin II (10 nmol) were also determined. RESULTS: The ICV injection of urotensin II at a dose of 1 nmol did not alter the arterial pressure or heart rate significantly, while 10 nmol urotensin II increased the arterial pressure and heart rate. The mean arterial pressure at 5 min of ICV urotensin II was 121 +/- 4 mmHg, which was significantly higher than that obtained by ICV injection of artificial cerebrospinal fluid (107 +/- 3 mmHg, P <0.05). In addition, significant increases in heart rate were observed 5-15 min after ICV urotensin II. Pre-treatment with pentolinium (5 mg/kg, i.v.) significantly attenuated the increases in mean arterial pressure (20 +/- 3 versus 8 +/- 2 mmHg, P <0.01) and heart rate (78 +/- 18 versus 7 +/- 5 beats/min, P <0.05) induced by ICV urotensin II. On the other hand, i.v. injection of urotensin II (10 nmol) elicited a depressor response associated with tachycardia; mean arterial pressure 5 min after injection was significantly lower in the urotensin II-injected rats (89 +/- 5 mmHg) than in the control rats (102 +/- 2 mmHg, P <0.05), and the heart rate was significantly higher in the former (402 +/- 11 versus 360 +/- 9 beats/min, respectively, P <0.05). CONCLUSIONS: Central urotensin II produces pressor and tachycardic responses through sympathetic activation, while peripheral urotensin II exerts a vasodilation-mediated depressor response in conscious rats. PMID- 12544447 TI - Nitric oxide production decreases after salt loading but is not related to blood pressure changes or nitric oxide-mediated vascular responses. AB - BACKGROUND: Nitric oxide production is a homeostatic mechanism that may regulate blood pressure during salt loading. Salt-sensitive hypertension in animal models and in humans is characterized by increased blood pressure and decreased nitric oxide production after salt loading. It is not known if this impaired nitric oxide production is the result of hypertension or is a mechanism contributing to the blood pressure response to salt. METHODS AND RESULTS: The effects of salt loading on blood pressure, nitric oxide-mediated vasodilation and nitric oxide production were measured in 25 normotensive subjects after 6 days on either a high (400 mmol/day) or low (10 mmol/day) sodium, low nitrate diet. Mean arterial pressure increased during the high-salt diet [4 +/- 1 mmHg (mean +/- SEM)] in 12 subjects and remained unchanged or decreased (-4 +/- 1 mmHg) in 13 subjects. Plasma nitrite and nitrate, a measure of nitric oxide production, decreased significantly from 39 +/- 3.3 micromol/l during the low-salt diet to 22.4 +/- 2.4 micromol/l during the high-salt diet (P = 0.0001). However, changes in mean arterial pressure from low- to high-salt diet did not correlate with changes in plasma nitrite and nitrate (r = 0.14, P = 0.51). Forearm blood flow increased significantly (P <0.0001) in response to mental stress, a nitric oxide-mediated response, but was not affected by sodium intake (from 7.8 +/- 0.9 to 11.2 +/- 1.4 ml/min per 100 ml during low salt versus 8.5 +/- 1.2 to 10.4 +/- 1.3 ml/min per 100 ml during high salt,P = 0.3). CONCLUSIONS: Salt loading results in a decrease in nitric oxide production in both salt-sensitive and salt-resistant normotensive subjects, which is independent of changes in blood pressure and does not affect the nitric oxide-mediated vascular response to mental stress. In contrast to salt resistant animal models, salt loading in healthy subjects does not increase nitric oxide production. Therefore, the increased blood pressure response to salt loading may occur through mechanisms other than nitric oxide, or salt-sensitive individuals are more sensitive to the reduced nitric oxide production that occurs after salt loading in both salt-sensitive and salt-resistant subjects. PMID- 12544449 TI - Importance of imidazoline-preferring receptors in the cardiovascular actions of chronically administered moxonidine, rilmenidine and clonidine in conscious rabbits. AB - OBJECTIVE: To determine the involvement of central imidazoline receptors in the cardiovascular actions of the chronically administered antihypertensive agents moxonidine, rilmenidine and clonidine. DESIGN AND METHODS: In 21 rabbits with implanted fourth-ventricular catheters, we investigated the central effects of three cumulative doses of an I(1)-imidazoline/alpha(2)-adrenoceptor antagonist, efaroxan, and of an alpha(2)-adrenoceptor antagonist, 2-methoxyidazoxan (2-MI), on the changes in blood pressure and heart rate (HR) elicited by chronic subcutaneous administration of moxonidine, rilmenidine and clonidine, after 1 and 3 weeks of treatment. A low, medium and high dose of 2-MI was matched to three doses of efaroxan, such that each produced equal reversal of the hypotension induced by fourth-ventricular alpha-methyldopa and hence produced a similar degree of alpha(2)-adrenoceptor blockade. RESULTS: Clonidine and moxonidine, at doses of 1 mg/kg per day, and rilmenidine at 5 mg/kg per day, produced sustained reductions in mean arterial pressure of 13 +/- 3, 15 +/- 2 and 13 +/- 2 mmHg, respectively over the 3-week treatment period, but did not alter HR. Central administration of efaroxan on day 9 and day 23 of treatment produced a greater increase in blood pressure than did 2-MI with all three antihypertensive agents. Blood pressure reached levels that were significantly above the original control values. By contrast, the alpha(2)-adrenoceptor antagonist 2-MI only induced a rebound blood pressure effect in clonidine- and to a lesser extent in rilmenidine treated rabbits. Both efaroxan and 2-MI produced a similar degree of tachycardia in moxonidine-, rilmenidine- and clonidine-treated animals.(2) CONCLUSIONS: The greater effect of efaroxan compared to the alpha(2)-adrenoceptor antagonist 2-MI suggests that the hypotension induced by chronic subcutaneous administration of moxonidine, rilmenidine and clonidine is mediated predominantly via an action on central imidazoline receptors. Furthermore, all agents showed a propensity to produce rebound hypertension with imidazoline receptor blockade. However, only clonidine showed a rebound phenomenon when challenged by acute central alpha(2) adrenoceptor blockade PMID- 12544450 TI - Chronic antioxidant treatment improves sympathetic functions and beta-adrenergic pathway in the spontaneously hypertensive rats. AB - BACKGROUND: Correlations have been found between oxidative stress and hypertension. OBJECTIVE: To determine whether antioxidants can normalize sympathetic dysfunction at pre- or postsynaptic levels in spontaneously hypertensive rats (SHRs). METHODS AND RESULTS: Untreated SHRs and Wistar-Kyoto (WKY) rats were compared with rats treated with melatonin (30 mg/kg per day) or N acetylcysteine (NAC) (4 g/kg per day) given in drinking water for 4 weeks. At the presynaptic level, SHRs had greater plasma noradrenaline concentrations (P <0.01) and an enhanced release of [3H]noradrenaline from isolated atria (P <0.001). At the postsynaptic level, they exhibited an increased proportion of beta2 adrenoceptors in the heart (P <0.001)and a decrease in the chronotropic and mean arterial pressure (MAP) responses to isoproterenol (P <0.001). Melatonin and NAC decreased MAP (P <0.001) and heart rate (P <0.05), and restored the plasma noradrenaline concentrations (P<0.01 and P <0.001, respectively), the chronotropic response to isoproterenol (P <0.05) and the proportions of beta(1)/beta(2)-adrenoceptors in the heart (P <0.05) in SHRs to the levels found in WKY rats. The same treatments decreased the release of [3H]noradrenaline from isolated atria (P <0.05), and melatonin slightly improved the relaxation in the aorta in SHRs only (P <0.05). Plasma concentrations of adrenaline, the isoproterenol-induced relaxation in mesenteric arteries, the total density and affinity of beta-adrenoceptors in the heart, and the adenylate cyclase reactivity of cardiac membranes to isoproterenol, forskolin, sodium fluoride and guanylylimidophosphate were not altered by the treatments. CONCLUSION: These results suggest that NAC and melatonin decreased the MAP and heart rate and improved the chronotropic response to isoproterenol in SHRs, in association with an inhibition of sympathetic activity and the restoration of cardiac beta adrenoceptor function. PMID- 12544451 TI - Resistance artery mechanics and composition in angiotensin II-infused rats: effects of aldosterone antagonism. AB - BACKGROUND: Angiotensin (Ang) II stimulates aldosterone production, which may mediate some of the effects of Ang II. OBJECTIVE: To test whether Ang II-induced structural and mechanical changes in resistance arteries may be prevented by the non-selective aldosterone receptor blocker, spironolactone, independently of reduction in blood pressure. METHODS: Male Sprague-Dawley rats received Ang II [120 ng/kg per min subcutaneously (s.c.)] with or without spironolactone or hydralazine (25 mg/kg per day). Two additional groups received aldosterone (750 ng/h s.c.) with or without spironolactone. After 2 weeks, third-order mesenteric arteries were dissected and studied by pressurized myograph. Deposition of collagen type I/III in the vascular wall was evaluated by confocal immunofluorescence microscopy. RESULTS: Ang II increased blood pressure significantly (P <0.01); this was partially prevented by spironolactone (P <0.01) and nearly normalized by hydralazine (P <0.01). Media thickness, media:lumen ratio and media cross-sectional area of mesenteric resistance arteries increased under Ang II or aldosterone (P <0.01) and this was partially prevented by spironolactone (P <0.01), but not by hydralazine. Compared with the control or Ang II + spironolactone groups, rats treated with Ang II with or without hydralazine presented stiffer vessels, with leftward shift of the stress-strain relationship and a raised slope of the incremental elastic modulus-stress relationship (P <0.05). Confocal microscopy demonstrated enhanced deposition of collagen type I/III in the media of arteries from rats infused with Ang II or aldosterone, an effect that was prevented partially by spironolactone but unaffected by hydralazine. CONCLUSION: Ang II-induced vascular alterations in structure, mechanics and composition were partially prevented by spironolactone, independently of blood pressure reduction, providing further evidence that some actions of Ang II on resistance arteries are mediated by aldosterone. PMID- 12544452 TI - Differential expression of D2-like dopamine receptors in the kidney of the spontaneously hypertensive rat. AB - OBJECTIVE: To compare the expression and cellular distribution of D(2)-like dopamine receptors in the kidney of the spontaneously hypertensive rat (SHR) and normotensive Wistar-Kyoto (WKY) rat. DESIGN: Renal D(2)-like receptor protein expression and distribution has not been studied in the SHR. Since changes in D(2)-like receptor expression and/or distribution may contribute to the dysregulation of renal dopamine and D(1A) receptor function, we examined the expression of the three subtypes of D(2)-like receptors (D(2), D(3) and D(4)) in SHR and WKY rat kidneys. METHODS: Western blot analysis and confocal immunocytochemistry with specific polyclonal antipeptide antibodies directed against the receptor subtypes, were used to assess protein expression. RESULTS: There were no differences in protein expression and cellular immunolocalization of the D(2) receptor subtypes between SHR and WKY rats. Expression of the 50 kDa D(3) receptor was reduced in the cortex of the SHR; no differences in D(3) receptor levels were seen in the inner medulla of SHR and WKY rats. The D(4) receptor polypeptides were overexpressed in the cortex of SHR, while in the inner medulla no difference in expression of the D(4) receptor proteins was observed between SHR and WKY rats. Immunocytochemistry also showed increased immunostaining of D(4) receptors in tubular structures in the cortex, but diminished staining in the SHR inner medulla. CONCLUSION: The observed differences in expression and distribution of D(3) and D(4) dopamine receptors between cortex and inner medulla of the kidneys of SHR and WKY rats may contribute to the aberrant state of dopaminergic-mediated natriuresis in SHR. PMID- 12544454 TI - Regression of left ventricular hypertrophy with echocardiography: some lessons from the LIVE study. AB - OBJECTIVE: Large trials on left ventricular hypertrophy (LVH) regression are indispensable to allow accurate evaluation of the different classes of treatments. Such a trial, the LIVE study, gave us the opportunity to examine the influence of the mode of reading of the echo tracings on results and to underline the value of two initial recordings performed at entry. METHODS: The LIVE study was designed to compare 1.5 mg indapamide SR with 20 mg enalapril on the regression of LVH evaluated echocardiographically. M-mode tracings of the left ventricle were performed at selection and after a 2 week placebo run-in. Recordings were read by investigators and by three experts all through the study, and finally read again by the three experts blind to treatment and sequence. RESULTS: The two initial examinations produced an estimate of the reproducibility of the measurement of the left ventricular mass (LVM) that can be proposed as an indicator of overall quality provided no patients are excluded. The standard deviation of the differences between these two estimates of LVM was 52 g. These two examinations may also help quantify regression to the mean, which was not significant for the whole group. Knowledge of the sequence of examinations overestimates the change in LVM with treatment, as shown by comparison of measurements carried out during quality control with the measurements made blind (-19 +/- 52 versus -6 +/- 53 g, P <0.01). CONCLUSIONS: The comparison of two initial echoes at entry allow one to define a quality criteria for such trials and to quantify a possible regression to the mean. The reading of recordings with knowledge of sequence significantly overestimates LVM variations on treatment. PMID- 12544455 TI - Dementia: whose problem is it, anyway? PMID- 12544453 TI - Disparate effects of angiotensin II antagonists and calcium channel blockers on albuminuria in experimental diabetes and hypertension: potential role of nephrin. AB - OBJECTIVE AND DESIGN: To explore the effects of various antihypertensive regimes which achieve similar blood pressure control using a range of agents including the angiotensin II type 1 receptor antagonist, valsartan, as monotherapy or in combination with two subclasses of calcium channel blockers (CCBs) (the dihydropyridine, amlodipine and the phenylalkylamine, verapamil) on the progression of renal disease and the expression of the podocyte slit pore protein, nephrin in an accelerated model of diabetic nephropathy. RESULTS: Valsartan treatment reduced systolic blood pressure as assessed by radiotelemetry (135 +/- 3 versus diabetic 153 +/- 6 mmHg) as well as retarding the increase in albumin excretion rate by approximately 50%. Combination therapy of valsartan with either amlodipine or verapamil was equally effective in reducing blood pressure to valsartan monotherapy (valsartan + amlodipine 129 +/- 4 valsartan + verapamil 133 +/- 6 mmHg;) but was not as effective at reducing albuminuria. A reduction in glomerulosclerosis was observed with valsartan monotherapy with less reduction in injury with the valsartan + amlodipine combination, despite a similar reduction in blood pressure. The decrease in nephrin, in diabetic rats was attenuated by valsartan monotherapy, but not by other treatments. CONCLUSIONS: The results of this study demonstrate that despite a similar reduction in blood pressure, the addition of the CCB amlodipine to the AII antagonist failed to provide similar renoprotection to that observed with an equihypotensive regimen of valsartan as monotherapy. Furthermore, the depletion in glomerular nephrin expression in diabetic animals was only abrogated by valsartan treatment, the therapy which was most effective at retarding the development of albuminuria in this model. PMID- 12544456 TI - What's new in Alzheimer's disease? AB - Major changes in the care and treatment of dementia of the Alzheimer's type have been made, and more are looming on the horizon. This article presents current information regarding pathology, diagnosis, causes, and treatment options. Additionally, the newest medications and alternative approaches to this prevalent disease are discussed. PMID- 12544457 TI - The importance of touch for the patient with dementia. AB - Massage has many benefits including general relaxation and increased circulation. Many experience pain relief from massage and may need less medication. This article describes how nurses and families can learn to provide touch therapy to patients with varying stages of dementia. PMID- 12544458 TI - Addressing the spiritual & religious needs of persons with profound memory loss. PMID- 12544459 TI - Music for your practice. PMID- 12544460 TI - A pain assessment tool for people with advanced Alzheimer's and other progressive dementias. AB - Appropriate pain management can only be achieved through accurate pain assessment that is individualized, ongoing, and well documented. Assessment tools must focus on the patient as the authority on pain's existence and severity; however, self reports are not feasible when patients lose their ability to verbally communicate. This article describes a scientifically proven pain assessment tool that can be used for patients with advanced dementia and Alzheimer's Disease. PMID- 12544461 TI - Techniques for bathing without a battle. PMID- 12544462 TI - Dignity and dietary interventions for dementia. PMID- 12544463 TI - Understanding hypersexuality: a behavioral disorder of dementia. AB - Although a rare manifestation of Alzheimer's Disease, hypersexuality is one of the most embarrassing behaviors for both informal and formal caregivers. This article presents an overview of the causes of the problem along with strategies home care and hospice nurses can use to teach families how to decrease the social isolation these patients experience. PMID- 12544464 TI - Benefits of pet therapy in dementia. AB - Pet therapy is a centuries-old treatment approach. The new news is that pet therapy is now being validated via scientific and anecdotal data proving its effectiveness. This article shows that pet therapy is worth considering for the care plan of persons with dementia. PMID- 12544468 TI - Caregiver role satisfaction: my mother's final gift. PMID- 12544465 TI - Palliative care for persons with dementia. AB - Home care and hospice nurses provide invaluable care for patients with Alzheimer's Disease and related disorders. Often these patients have advanced disease and may be approaching the end of life. This article describes a palliative care approach to providing comprehensive assessment, care planning, and interventions enhancing the functioning and quality of life for both the patient and family coping with this illness. PMID- 12544469 TI - Hereditary hemochromatosis: perspectives of public health, medical genetics, and primary care. AB - Hereditary hemochromatosis (HHC) is a condition characterized by excess iron in body tissues, resulting in complications such as cirrhosis, cardiomyopathy, diabetes, and arthritis. These complications usually manifest during adulthood. Two methods of screening for the detection of early stage of HHC are available: serum iron measures and molecular testing to detect mutations in the gene. These phenotypic and genotypic screening tests are of particular interest because a simple treatment-periodic phlebotomy-can be used to prevent iron accumulation and clinical complications. HHC might represent the first adult-onset genetic disorder for which universal population-based screening would be appropriate. Therefore, HHC has been proposed as a paradigm for the introduction of adult genetic diseases into clinical and public health practice. However, universal screening for HHC has not been recommended because of the uncertainty about the natural history of the iron overload or HHC and, in particular, uncertainty about the prevalence of asymptomatic iron overload and the likelihood that it will progress to clinical complications. If universal screening is not appropriate based on current data, what other measures might reduce the disease burden of iron overload? New studies provide more systematic information about the penetrance of the C282Y mutation and shed further light on the natural history of the disorder. The authors review these data and consider their implications for public health, medical genetics, and primary care. PMID- 12544470 TI - Extensive sequencing of the cystic fibrosis transmembrane regulator gene: assay validation and unexpected benefits of developing a comprehensive test. AB - PURPOSE: To develop a sequencing assay for the gene to identify mutations in patients with cystic fibrosis (CF). METHODS: An automated assay format was developed to sequence all exons and splice junctional sequences, the promotor region, and parts of introns 11 and 19. RESULTS: After validating the assay using 20 known samples, DNA of seven patients, four of whom were heterozygous for a known CF mutation, was sequenced. Known CF mutations were detected in seven of the eight chromosomes, and a novel missense mutation was detected in the eighth. In addition, this assay allowed 14 ambiguous results obtained using the Roche CF gold strips to be resolved. Three false-positive diagnoses were prevented; a different mutation at the same codon was identified in two patients and confirmation was provided in the remaining nine cases. CONCLUSIONS: Sequencing of the gene provides important information for CF patients and is a valuable adjunct to a carrier screening program to resolve ambiguities in panel testing. PMID- 12544471 TI - Analytic validity of cystic fibrosis testing: a preliminary estimate. AB - PURPOSE: Derive estimates of analytic sensitivity and specificity of DNA testing for cystic fibrosis in the United States. METHODS: Analyze published results of the American College of Medical Genetics (ACMG)/College of American Pathologists (CAP) Molecular Genetics Survey between 1996 and 2001, taking into account difficult, simulated clinical samples included for educational purposes. RESULTS: Analytic sensitivity is 97.9% [95% confidence interval (CI) 96.8-98.7%], and analytic specificity is 99.4% (95% CI 98.7-99.9%) after removing challenges involving delI507 and performing other adjustments. Analytic sensitivity is consistent over the 6 years. Specificity was lower in 1997. CONCLUSION: These preliminary estimates indicate that analytic validity of cystic fibrosis testing is very good and can likely be improved. To date, fewer than half of the mutations in the panel recommended for preconceptional or prenatal screening have been challenged. The present study highlights the value of performing confirmatory testing when a mutation is identified to reduce false-positive results. PMID- 12544472 TI - The Stickler syndrome: genotype/phenotype correlation in 10 families with Stickler syndrome resulting from seven mutations in the type II collagen gene locus COL2A1. AB - PURPOSE: To evaluate a cohort of clinically diagnosed Stickler patients in which the causative mutation has been identified, determine the prevalence of clinical features in this group as a whole and as a function of age, and look for genotype/phenotype correlations. METHODS: Review of medical records, clinical evaluations, and mutational analyses of clinically diagnosed Stickler patients. RESULTS: Patients with seven defined mutations had similar phenotypes, though both inter- and intrafamilial variability were apparent and extensive. The prevalence of certain clinical features was a function of age. CONCLUSION: Although the molecular determination of a mutation can predict the occurrence of Stickler syndrome, the variability observed in the families described here makes it difficult to predict the severity of the phenotype on the basis of genotype. PMID- 12544473 TI - Utility of subtelomeric fluorescent DNA probes for detection of chromosome anomalies in 425 patients. AB - PURPOSE: A complete set of subtelomeric fluorescent DNA probes, except the acrocentric p-arms, was developed in 1996, was optimized in 1998, and is commercially available. These and other fluorescence in situ hybridization (FISH) probes have been used to detect anomalies of the subtelomere regions among groups of patients with idiopathic mental retardation (MR), developmental delay (DD), and/or nonspecific dysmorphic features (NDF), and individuals with multiple miscarriages (MM) who were karyotypically normal by standard G-banding techniques. METHODS: A total of 425 patients were analyzed, of whom 372 had idiopathic MR/DD/NDF and 53 were involved in MM. An effort was made to select individuals for this study who were either normal karyotypically or who had subtle chromosomal anomalies that were inconclusive by banded chromosome analysis, although this was not always possible. RESULTS: Anomalies involving the subtelomere regions were detected at a frequency of 6.8% in the MR/DD/NDF group. The cryptic or subtle anomalies are estimated to be about 3.4%. It was necessary to use M-FISH, chromosome, and locus specific FISH probes to clarify some of the abnormalities. No abnormalities were detected in the MM group. Deletion variants were present for 2qter, 7pter, and Xpter/Ypter subtelomeric regions ranging from <1 to 9.6%. CONCLUSIONS: The subtelomeric FISH probes are instrumental in the detection of subtelomeric anomalies in a significant proportion, although no more than 50% are subtle, of patients with idiopathic MR/DD/NDF. In some cases, however, it was necessary to use other FISH probes to clarify the nature of these abnormalities. No subtelomeric abnormalities were detected in our group of 53 MM patients, suggesting a relatively low frequency of occurrence in this patient population. PMID- 12544474 TI - Consent for genetic research in a general population: the NHANES experience. AB - PURPOSE: To determine the sociodemographic factors associated with consent for storage of DNA for future genetic research. METHODS: Analysis of the characteristics of consenting individuals participating in the National Health and Nutrition Examination Survey, a nationally representative survey of the US household population. RESULTS: In 1999, 84% (95% confidence interval 82.4-85.6) of eligible participants consented to have their blood samples included in a national repository for genetic research. In 2000, 85.3% (95% confidence interval 84.0-86.6) consented. Females and black participants in both years were least likely to consent (1999, 82.2% and 73.2%; 2000, 83.6% and 81.3%, respectively). An assessment by logistic regression demonstrated that in both years only non Hispanic black race/ethnicity was a significant independent predictor for not consenting to future genetic research. CONCLUSION: Although non-Hispanic black individuals have overall response rates similar to those of the other racial/ethnic groups, they are less likely to agree to have a blood sample saved for future genetic research. In balance, however, these findings demonstrate wide acceptance among survey participants for allowing storage of specimens for future genetic research across many demographic variables. PMID- 12544475 TI - Teaching clinical management skills for genetic testing of hereditary nonpolyposis colorectal cancer using a Web-based tutorial. AB - PURPOSE: To pilot and evaluate an interactive Web-based continuing medical education tutorial on clinical management of hereditary nonpolyposis colon cancer (HNPCC) and genetic testing. METHODS: Gastroenterology fellows and genetic counseling trainees were asked to read standard written materials before taking the tutorial. A pretest/post-test assessment was used to measure change in subjects' clinical management skills. RESULTS: Subjects made the correct management decision 63.9% of the time before the tutorial and 81.1% of the time after the tutorial (P < 0.001). CONCLUSIONS: Supplementing written materials with an interactive program may assist medical professionals in integrating their knowledge of HNPCC and genetic testing into clinical practice. PMID- 12544476 TI - Attitudes of African American premedical students toward genetic testing and screening. AB - PURPOSE: Genetic research is progressing at a rapid rate. While most view genetic advances favorably, concerns regarding eugenics and discrimination based on genetic test results have been raised. These concerns have been found among all groups studied; however, they have particular relevance for members of the African American community. Studies have shown that because of a long history of negative experiences, African Americans have a general mistrust of the medical establishment. It is unclear whether these negative attitudes encompass genetic advances. Because there is little empiric data in the literature, it is not known whether African Americans have a positive view of genetic advances or whether they have the same level of mistrust as is seen in their attitudes toward other forms of biomedical research. METHODS: This study was conducted as an initial effort to examine the attitudes of African Americans toward recent genetic advances and, specifically, genetic testing. A cohort of 97 college-age minority students, including 78 African Americans, participating in the Health Career Enhancement for Minorities Program (HCEM) at Case Western Reserve University were surveyed. Surveys were made available before and after the summer long course, which included five lectures on basic genetic principles and medical genetics. RESULTS: Both African American students and other minority students initially (questionnaire prior to HCEM course) had an overall positive view of genetic testing. The vast majority supported genetic testing for preventive care (95%) and presymptomatic detection of disease (88%) and agreed that it should be easily available (83%). However, several concerns were expressed as well, including fears about discrimination (68%), privacy (68%), that abortions will become more common (51%), and eugenics (37%). It is interesting that in the postcourse questionnaire, the percentages of positive views remained similar to those of the precourse survey, but the number of respondents expressing concerns increased. DISCUSSION: These results suggest that the minority students surveyed view many aspects of genetic testing and other advances favorably. However, these students expressed concerns about discrimination, privacy, and eugenics. These concerns were increased, not lessened, by exposure to genetics education. One possible explanation for this observation is that the students had a greater understanding of the issues regarding genetic testing after the HCEM lectures and discussion. Of note, there was a greater negative response toward genetic screening programs among the African American students compared with the non-African American minority students. This suggests that the negative attitudes of African Americans toward biomedical research do extend to some aspects of genetics and that educational programs must be designed and implemented if this community is going to receive the maximum benefits of this advancing technology. PMID- 12544477 TI - The genetics workforce and workload. PMID- 12544478 TI - Tetrahydrobiopterin control in phenylketonuria. PMID- 12544480 TI - Genomics, proteomics, and the new paradigm in biomedical research. AB - This article is based on the keynote address that introduced the third biennial Asan Medical Center-Harvard Medical International Symposium "Genetics and Proteomics: Impact on Medicine and Health" that took place in Seoul, Korea, July 3-4, 2001. In his address, the author summarized exciting achievements in the field of genomics and introduced the related and emerging field of proteomics. By using industrialized high-throughput approaches, genomics and proteomics are dramatically accelerating the pace of biological research. They have started a scientific revolution whose impact will range from elucidating the structure of our chromosomes to providing powerful new tools for the study of disease; and from understanding human evolutionary history to novel applications in the medicine of the future. The author's overview highlighted the recent history of the two fields and laid the foundation for the rest of the symposium presentations. PMID- 12544481 TI - Genetics in medical practice. AB - Medical genetics has been formally recognized as a medical specialty in the United States only within the past decade. Initially, medical genetics was concerned with relatively rare single gene or chromosomal disorders, but with the sequencing of the human genome, genetics has become the driving force in medical research and is now poised for integration into medical practice. This article offers a perspective on the role of genetics in medical practice and how this role may evolve over the next several years. The author classifies the genetic contribution to medical practice into three categories: monogenic or chromosomal disorders, more common disorders due to a monogenic or single gene mutation, and common multifactorial disorders in which the interaction of multiple genes and the environment contribute to the cause of the condition. The author discusses the positives and the potential pitfalls facing primary care physicians and specialists as they incorporate medical genetics into their practices and points out some of the anticipated changes medical genetics will necessitate, such as an increased medical genetics workforce, including geneticists and genetic counselors. The Human Genome Project has made information of inestimable diagnostic and therapeutic importance available. The medical profession now must rise to both the opportunities and challenges that this wealth of information presents. PMID- 12544482 TI - Genetic advances will influence the practice of medicine: examples from cancer research and care of cancer patients. AB - Exciting new techniques of genomics, proteomics, and bioinformatics are beginning to influence the practice of medicine, most notably in diagnosis and drug development for patients with various cancers. Examples are drawn from B-cell lymphomas, melanomas, and prostate, lung, and breast cancers. As in all evidence based clinical practice, physicians will be better prepared if they understand the nature of the tests and the kinds of information from which they and their consultants will make clinical inferences and assist patients in making clinical decisions. Physicians also can help put new technologies in cultural and ethical context. PMID- 12544483 TI - The crucial role of the public health sciences in the postgenomic era. AB - Genomic sequence information and gene and protein expression patterns must be linked with information about diet and metabolism, lifestyle behaviors, diseases and medications, and microbial, chemical, and physical exposures. These linkages depend upon the public health sciences: epidemiology, biostatistics, environmental health sciences, pathobiology, health sciences research, and clinical prevention trials. PMID- 12544484 TI - Reflections on the ethics of genetic enhancement. AB - The technologies made available by new research in genetics and genomics have been and will be used not only to diagnose and treat disease, but also to attempt to enhance human traits and capacities. A broad definition of genetic-enhancement technologies, not merely gene manipulation, but also indirect genetic technologies, such as biosynthetic drugs, is needed to capture the full range of possible applications. It is difficult, but often possible, to anticipate the enhancement temptations of new therapies. Such anticipation may help us in shaping the marketing, availability, or other aspects of those technologies. Appropriate public and professional policies are needed, and work on them should begin immediately. Most important, we must stimulate public education and dialogue to encourage personal ethical reflection on the appropriate uses and limits of genetic-enhancement technologies. PMID- 12544485 TI - Integration of genetics into clinical teaching in medical school education. AB - Medical genetics has moved from the study of rare conditions to the illumination of disorders that impact the entire spectrum of medical practice. While there have been a number of predictions and concerns about this impact, this article examines three areas where medical genetics is clearly an important tool in medical practice. First, a family history aids in risk assessment, even in common disorders that are multifactorial. Second, by elucidating molecular pathways, gene identification may lead to the development of more efficacious medications that have fewer side effects. Third, an awareness of population-based risk and the availability of genetic screening in these populations will help physicians assess an individual patient's risk. To fully benefit from genetically based medical approaches, physicians will need to master a new set of principles and clinical skills. However, genetics has traditionally been taught as a basic science, sometimes under the purview of cell biology or biochemistry. Often, then, genetics has little or no place in clinical teaching. This article describes an effort at Harvard Medical School to integrate genetics into both the preclinical and the clinical curricula. The author looks at the underlying pedagogy, how basic science teaching in genetics is provided, and an approach currently being used to include genetics in clinical teaching. PMID- 12544486 TI - Coupling genomics and human genetics to delineate basic mechanisms of development. AB - This article, based on a presentation given by the author at the third Asan Harvard Medical International Symposium on "Genomics and Proteomics: Impact on Medicine and Health" in Seoul, Korea, July 3-4, 2001, discusses an iterative translational research approach to delineate the basic mechanism of human development. The study of humans to increase the understanding of mammalian development has critical advantages that make its limitations acceptable for certain types of studies. For instance, by looking at families affected by birth defects, researchers can gain insight into the basic mechanisms of development and how genes program organisms to assume their permanent, or adult, morphological shapes. A number of malformation syndromes have some overlapping manifestations, despite being phenotypically and, in some cases, genetically distinct. What can researchers learn from this? The author's research group clinically and genetically analyzed families affected with the Pallister-Hall syndrome. The researchers then went on to look at the McKusick-Kaufman syndrome, a disorder that is more common among the Old Order Amish of Lancaster County, Pennsylvania, in an attempt to understand more about genes, genetic pathways, and syndrome families. PMID- 12544487 TI - Identification of novel mutations and the three most common mutations in the human ATP7B gene of Korean patients with Wilson disease. AB - PURPOSE: Wilson disease, an autosomal recessive disorder of copper transport, is probably the most common inherited metabolic disorder in Korea. In Wilson disease, synthesis of a defective copper transporting enzyme leads to the accumulation of copper in the liver, brain, and kidney. The product of the Wilson disease gene is a copper transporting P-type ATPase (ATP7B). In this study, efforts were made to identify novel mutations and investigate the frequency of the common mutations in Korean patients with Wilson disease. METHODS: This study includes 37 patients from 33 unrelated Korean families with Wilson disease. Genomic DNA from peripheral leukocytes or skin fibroblasts and cDNA from liver tissue were polymerase chain reaction-amplified exon by exon and subsequently analyzed using heteroduplex or single-strand conformation polymorphism analysis. Specimens showing mobility shift on those studies were directly sequenced. RESULTS: Twelve different mutations in 33 Korean families with Wilson disease were identified: Arg778Leu (R778L), Asn1270Ser (N1270S), Ala874Val (A874V), 2303 2305delC, 2630-2656del, 2460-2462insC, Cys656Stop (C656X), Pro768His (P768H), Leu1083Phe (L1083F), Ala1168Ser (A1168S), Leu1255Ile (L1255I), and Asp1267Ala (D1267A). Among these, six mutations (2630-2656del, 2460-2462insC, C656X, P768H, A1168S, and L1255I) are novel. The R778L mutation is known to be highly prevalent in Asian patients. The allele frequency of R778L in Korean patients with Wilson disease was 37.9%, which was significantly higher than those of Japanese and Taiwanese patients. The N1270S mutation, originally described in an Italian patient, was the next most common mutation in Korean patients, with an allele frequency of 12.1%, which was presumed to disrupt the ATP hinge domain of the ATP7B protein. The A874V mutation was the third most common mutation with an allele frequency of 9.4%, which was presumed to disrupt the Td domain of the ATP7B protein. CONCLUSION: The R778L, N1270S, and A874V mutations are three major mutations representing approximately 60% of mutated alleles, although Korean patients with Wilson disease are genetically heterogeneous. PMID- 12544488 TI - Allele frequency determination of publicly available cSNPs in the Korean population. AB - PURPOSE: As a first step toward the construction of a single-nucleotide polymorphism (SNP) database of the Korean population, the authors determined the allele frequencies of 406 cSNPs selected from the public database. METHODS: A pooled DNA sequencing approach was used to determine the allele frequencies of 406 cSNPs selected from 120 genes in 24 individuals. RESULTS: Of 406 cSNPs, 53% were monomorphic in the Korean samples. Among tested SNPs, 292 SNPs (72%) were uncommon (minor allele < 20%) and 114 SNPs (28%) were common (minor allele > or = 20%) in our population. CONCLUSION: An extensive SNP characterization would be necessary, and the ethnic and population-based differences should be considered in the selection of SNPs for the study of complex diseases with association mapping methods. PMID- 12544489 TI - Progress and limitations in cancer gene therapy. AB - This is a brief discussion on the progress of gene therapy and the limitations of present-day gene therapy clinical trials based on a review of 464 human trial protocols from the U.S. National Institutes of Health (NIH) and the U.S. Federal Drug Administration (FDA). The paper also discusses an aspect of the gene therapy research of the author, who, together with colleagues, conducted the first gene therapy clinical trial in Korea in 1995. PMID- 12544490 TI - AAV-mediated gene transfer for hemophilia. AB - The goal of our work has been to establish an experimental basis for gene transfer as a method of treating hemophilia, an inherited bleeding disorder that results from the absence of functional factor VIII or factor IX. Using an adeno associated viral vector derived from AAV serotype 2, we have shown in mice and in hemophilic dogs that we can achieve long-term expression (>3 years) of clotting factor at levels that would result in an improvement of clinical symptoms of the disease. A phase I trial of intramuscular injection of AAV-F.IX showed no evidence of local or systemic toxicity in any of the subjects. Muscle biopsies showed evidence for gene transfer and expression by polymerase chain reaction, Southern blot, and immunohistochemistry. We have also shown that AAV-F.IX can be delivered into the portal veins of hemophilic dogs and that this results in high circulating levels of factor IX, on the order of 5% to 14%, whereas delivery of similar doses to skeletal muscle results in factor levels of only 1% to 2%. Based on these results, a trial of AAV-mediated liver-directed gene transfer for hemophilia B has been proposed and is reviewed here. PMID- 12544491 TI - Bioinformatics and genomic medicine. AB - Bioinformatics is a rapidly emerging field of biomedical research. A flood of large-scale genomic and postgenomic data means that many of the challenges in biomedical research are now challenges in computational science. Clinical informatics has long developed methodologies to improve biomedical research and clinical care by integrating experimental and clinical information systems. The informatics revolution in both bioinformatics and clinical informatics will eventually change the current practice of medicine, including diagnostics, therapeutics, and prognostics. Postgenome informatics, powered by high-throughput technologies and genomic-scale databases, is likely to transform our biomedical understanding forever, in much the same way that biochemistry did a generation ago. This paper describes how these technologies will impact biomedical research and clinical care, emphasizing recent advances in biochip-based functional genomics and proteomics. Basic data preprocessing with normalization and filtering, primary pattern analysis, and machine-learning algorithms are discussed. Use of integrative biochip informatics technologies, including multivariate data projection, gene-metabolic pathway mapping, automated biomolecular annotation, text mining of factual and literature databases, and the integrated management of biomolecular databases, are also discussed. PMID- 12544492 TI - The human genome: cause to remember our humanity. AB - This article is based on the closing address given by Dr. Mitchell Spellman on behalf of Harvard Medical International at the third biennial Asan Medical Center Harvard Medical International Symposium titled "Genetics and Proteomics: Impact on Medicine and Health" that took place at the Asan Medical Center in Seoul, Korea, July 3-4, 2001. Dr. Spellman's remarks were addressed to a distinguished gathering that included President Changgi D. Hong and President Emeritus Pyung Chul Min of Asan Medical Center; President Emeritus Mun Ho Lee of Asan Medical Center, who is currently President, Korea Cancer Research Foundation; Secretary General Chul Lee of the Organizing Committee for the Symposium; Dr. Won Jong Kim, Dean of the University of Ulsan College of Medicine; Dr. Je Guen Chi, President of the Korean Academy of Medical Sciences; and the Faculty of the Symposium, together with colleagues, guests, and friends. First, Dr. Spellman reminds participants of the breadth of topics presented during the third biennial symposium. He then discourses on the inestimable worth of the sequencing of the human genome and how such knowledge provides us with the opportunity to consider the meaning and dimensions of humanity-biological, social, and ethical. PMID- 12544493 TI - Integrating genomics into US public health. AB - This paper presents an overview of how the US government is currently addressing the complex issue of integrating genomics into public health. The author, a public health provider with many years of experience, including several years spent in genetics and maternal and child health, is currently a senior advisor to the Surgeon General of the United States. At the time of the third biennial Asan Harvard Medical International Symposium, "Genomics and Proteomics: Impact on Medicine and Health," in Seoul, Korea, July 3-4, 2001, the US Surgeon General was Dr. David Satcher. Dr. Allan Noonan attended the symposium as Dr. Satcher's representative. Dr. Noonan's paper discusses the challenges facing the US public health system as it strives to integrate and promote genomics and proteomics into its efforts to reduce morbidity and mortality in the US population. PMID- 12544494 TI - Key roles of government in genomics and proteomics: a public health perspective. AB - This paper is based on the presentation of Dr. Allan Noonan at the third biennial Asan-Harvard Medical International Symposium on "Genomics and Proteomics: Impact on Medicine and Health" that took place in Seoul, Korea, July 3-4, 2001. Dr. Noonan is a senior advisor to the Surgeon General of the United States and was representing the then Surgeon General, Dr. David Satcher. In this final presentation of the symposium, Dr. Noonan reviews the key roles of government in US health care and discusses several areas where genomic- and proteomic-based information will necessitate changes in the functions of public health. In particular, Dr. Noonan discusses the need for appropriate training to meet the challenges of the genomic future; for sensitivity in the development of policies to address the ethical, legal, and social implications of genomic information; and for dissemination of genomic information to both the professionals and the public. Dr. Noonan concludes with a vision of the genomic future of the next 30 years and a reiteration of the need for partnership among health professionals, educators, and social services professionals. PMID- 12544495 TI - Breast cancer screening: controversies and future directions. AB - PURPOSE OF REVIEW: Recent criticisms of the mature breast cancer screening trials claimed that there is no evidence that screening saves lives. This has developed into a major public controversy, causing physicians, women, and policy analysts to rethink and debate mammography-screening guidelines. We have studied this subject from a different perspective -- using computer simulation to fit a simple growth model to clinical data. We can thus provide another viewpoint of the screening controversy that may help elucidate the underlying biology and aid policy makers in devising sound screening guidelines. RECENT FINDINGS: We agree with some reviewers that there is partial validity to the criticism. Based on our studies, we have arrived at a new explanation of why screening has not lived up to expectations. SUMMARY: Our fundamental hypothesis is that breast cancers often undergo periods during which they are temporarily dormant. In addition, surgical intervention to remove primary tumors can interrupt this dormancy. Therefore screening finds smaller tumors with fewer positive lymph nodes, which is beneficial. But then the resulting extirpation accelerates the growth of dormant distant micrometastases, and results in earlier relapses than in women who have not been screened. This partly offsets the early detection advantage. One hypothetical mechanism proposed to explain this biology is that surgical wounding, particularly for premenopausal node-positive patients, can trigger the angiogenesis of dormant avascular micrometastases. PMID- 12544496 TI - Local failure and prognostic factors in ductal carcinoma in situ: concentration on recent publications. AB - PURPOSE OF REVIEW: Traditional therapy for ductal carcinoma in situ has been mastectomy or conservative surgery plus radiation, but interest in more expeditious therapies such as conservative surgery alone is growing. A first step in considering changes in an already successful treatment is to evaluate tumor and patient factors that may predict outcome. Identifying and understanding these prognostic factors will aid researchers in designing new treatment paradigms. The purpose of this review is to assess the value of commonly accepted potential prognostic factors in patients with ductal carcinoma in situ. RECENT FINDINGS: During this past year several retrospective studies have evaluated putative prognostic factors in ductal carcinoma in situ. We will compare these later publications with results from prospective randomized trials and one meta analysis. SUMMARY: There does not appear to be unanimous agreement on the prognostic factors associated with increased risk of local failure in patients treated with conservative surgery and radiation for ductal carcinoma in situ. In our chosen papers, however, comedonecrosis and positive margins are most likely prognostic factors for increased local failure. PMID- 12544497 TI - Update on clinical role of tamoxifen. AB - PURPOSE OF REVIEW: Breast cancer is the most common malignancy amongst women in the United States and decreased mortality over the past decade has been attributed to a combination of screening and adjuvant therapies. There has been a resurgence of interest in hormonal therapies and this article discusses the clinical status of tamoxifen in the context of emerging alternative agents for treatment and prevention of breast cancer. RECENT FINDINGS: Tamoxifen has served as a prototype for the development of selective estrogen receptor modulators at the laboratory-clinical interface. Molecular technologies have permitted elucidation of mechanisms for tissue specific action and led to newer selective estrogen receptor modulators with potentially greater antitumour efficacy and attenuated uterotrophic profile. Publications over the past 12 months have emphasized the risks of thromboembolism and endometrial carcinoma associated with tamoxifen use which has accelerated application of other hormonal agents for treatment of advanced disease and as neoadjuvant therapy. This article reviews the current role of tamoxifen in the treatment of early and advanced breast cancer together with its potential for chemoprevention. Models for quantitative risk assessment are being developed to identify women for whom chemoprotection is justified. SUMMARY: Recent data showing a survival advantage for the aromatase inhibitor letrozole compared with tamoxifen in the advanced setting and improvement in disease-free survival for the aromatase inhibitor anastrozole versus tamoxifen as adjuvant treatment may herald a major shift in standard first line endocrine therapies for both advanced and early disease and ultimately chemoprevention of breast cancer. Other agents including newer SERMs and pure antiestrogens are undergoing phase III clinical trials and future endocrine and biological therapies are likely to include more selective and targeted therapies, which may be efficacious in both hormone-sensitive and receptor-negative disease. PMID- 12544498 TI - Neoadjuvant chemotherapy and surgical considerations in ovarian cancer. AB - PURPOSE OF REVIEW: Despite advances in surgery, it is still not possible in most patients with advanced ovarian carcinoma to remove the tumour completely. For these patients the concept of primary chemotherapy followed by interval debulking has emerged. Various studies in the past few years have evaluated the feasibility and benefit of this therapeutic approach. The available data is presented and discussed in this review. RECENT FINDINGS: The indication for interval surgery was generally based on the response to chemotherapy. However, different criteria of remission were adhered to, possibly explaining the varying outcomes of the trials. The right selection of patients suitable for this approach is crucial and needs further investigation. In these cases with an unfavourable prognosis, higher tumour resection rates and longer median survival times can be achieved by the use of neoadjuvant chemotherapy. SUMMARY: Until the results of a prospective randomized study become available, the use of neoadjuvant chemotherapy followed by debulking laparotomy must still be regarded as experimental, and must not be applied outside clinical trials. PMID- 12544499 TI - Update on granulosa cell tumours of the ovary. AB - PURPOSE OF REVIEW: Granulosa cell tumours of the ovary are an uncommon ovarian sex-cord stromal tumour. These neoplasms provide a spectrum of clinical presentations that span from the first to the tenth decade. Surgery represents the primary therapy for early stage disease; however, management of women with advanced disease is less clear. Because of their relative rarity, evidence to support decision-making in the management of granulosa cell tumours is limited. The purpose of this review is to provide the clinician with an updated knowledge of the clinical and molecular aspects of granulosa cell tumours in order to guide therapy. RECENT FINDINGS: The clinical stage, mitotic index and cellular atypia correlate most strongly with prognosis. However, these tumours may demonstrate heterogeneous genetic aberrations that can predict behaviour and response to therapy. Case series and reports suggest that postoperative combination chemotherapy is of most benefit in advanced disease. Serial measurements of serum inhibin may be helpful in the follow-up of these women, particularly in the post menopausal group. SUMMARY: The pathology and treatment of women with granulosa cell tumours of the ovary is complex. Such women should be managed in a multidisciplinary gynaecological oncology unit. A better understanding of the molecular pathology may assist treatment. PMID- 12544500 TI - Borderline ovarian tumours: a review of the recent literature. AB - PURPOSE OF REVIEW: Borderline tumors of the ovary are seen commonly in obstetrics and gynecology practices. This review will outline the most recent information regarding the epidemiology, molecular pathogenesis, pathology, and clinical management of these tumors. RECENT FINDINGS: In the past, borderline ovarian tumors and invasive ovarian cancer were studied together, under the presumption that they represent a continuum of disease. This view is coming into question based on both epidemiological and molecular biological studies. Pathologists are increasingly able to identify poor prognostic histological features. This has not yet been translated into improved non-surgical therapy. Surgical management to excise all visible tumor remains the cornerstone of therapy. Because borderline ovarian tumors often occur in reproductive-age women, fertility is an important issue. Conservative surgery is safe in carefully selected patients. SUMMARY: Our growing understanding of the relatively benign natural history of borderline ovarian tumors has allowed us to be more conservative in their surgical management, preserving fertility in many young women. A few women will be affected by a more virulent form of the disease with the potential to recur and cause death. The pathological features that identify patients with a poor prognosis are progressively being elucidated. Surgery remains the most effective therapy for these patients. Effective non-surgical therapies have yet to be identified. Hopefully, as molecular biological studies continue to expand our understanding of this disease as probably a separate carcinogenic process from invasive ovarian cancer, we will be able to employ this knowledge to design novel adjuvant therapies. PMID- 12544501 TI - Angiogenesis: prognostic and therapeutic implications in gynecologic and breast malignancies. AB - PURPOSE OF REVIEW: Growth, invasion and metastasis of malignant tumors depend on angiogenesis, which is therefore considered an attractive therapeutic target and indicator of clinical outcome. Summarizing the results of very recent studies, we discuss the clinical relevance of vascularization of breast and gynecological tumors. RECENT FINDINGS: The prognostic values of directly assessed vascularity (microvessel density) and angiogenic factors have been confirmed for these malignancies, despite few contradictory results that may be due to methodological inaccuracy. In addition, many studies point to a role of angiogenesis as a predictor of response to both antiangiogenic and conventional chemotherapy or radiotherapy. For a conclusive evaluation, however, further studies covering all clinically relevant subgroups of patients are needed. Novel therapeutic approaches with inhibitors of angiogenesis are promising, although the adaptation of most experimentally proven approaches to clinical practice appears to be a laborious process. Furthermore, for an appropriate clinical evaluation of these drugs, newly defined endpoints of treatment and new monitoring systems reflecting their mode of action and the anticipated biological effects are required. SUMMARY: The prognostic and predictive value of angiogenic factors and microvessel density has been confirmed. Antiangiogenic therapy will most likely play an important role in future clinical management of malignant diseases. PMID- 12544502 TI - New technologies for the identification of markers for early detection of ovarian cancer. AB - PURPOSE OF REVIEW: The ovarian cancer screening regimens in current use fail to identify disease at an early curable stage. RECENT FINDINGS: New technologies are emerging that facilitate the identification of diagnostic tumor markers. In particular, high throughput techniques using microarray technology and proteomic screening have enriched the study of protein expression by ovarian cancer cells. SUMMARY: Further evaluation of serum proteins associated with ovarian cancer holds promise for the development of a tumor marker panel that could aid in the early diagnosis of ovarian cancer, and save lives. PMID- 12544503 TI - Sexual dysfunction after treatment for gynaecologic and breast malignancies. AB - PURPOSE OF REVIEW: This review summarizes current knowledge about the nature of sexual dysfunction in gynaecologic and breast cancers, highlighting recent publications on treatment effects and communication issues. RECENT FINDINGS: In both gynaecologic and breast cancer, sexual dysfunction causes much distress to patients, from the time of diagnosis through to long-term follow-up. It appears that younger women in particular experience difficulties related to loss of reproductive function and relationship problems, plus more abrupt vaginal changes, than older women. Chemotherapy has been shown to be associated with short and long-term effects on sexual functioning and quality of life in breast cancer, and it is anticipated that this would extend to gynaecologic cancers also. The addition of endocrine treatments to chemotherapy in breast cancer appears not to affect levels of sexual functioning, although this may depend on the age of the woman. Sexual self-schema appears to be an important concept in predicting sexual dysfunction. Communication with women about sexual issues is vital, but evidence suggests this is lacking. SUMMARY: We are beginning to understand more about the sexual implications of different treatments and to identify factors which predict sexual dysfunction. More research is still required to identify how sexual dysfunction is affected in different groups of women and how best to help women who experience sexual difficulties. Recent findings suggest that there is a need to increase communication and support about sexual issues. PMID- 12544504 TI - Risk factors and risk reduction of breast and ovarian cancer. AB - PURPOSE OF REVIEW: Breast and ovarian cancer remain a significant burden for women living in the Western world. This paper reviews the risk factors and current strategies to prevent these diseases. RECENT FINDINGS: Established factors associated with the risk of breast cancer include family history, reproductive factors and lactation, as well as age at menarche and menopause. Hormone replacement therapy increases the risk, whereas oral contraceptives probably confer no increased risk. Alcohol moderately increases the risk, whereas a diet rich in folate and carotenoids might be protective. The role of other dietary factors, smoking and physical exercise remain unclear. Important risk factors for ovarian cancer are reproductive factors and possibly the long-term use of hormone replacement therapy. The risk is decreased by oral contraceptives. In carriers of a BRCA1 or BRCA2 gene mutation, prophylactic surgery can significantly reduce the risk of breast as well as ovarian cancer. Tamoxifen may be considered as a chemopreventive agent in women with a high risk of breast cancer, including carriers of a BRCA2 mutation, but is probably not effective in BRCA1 carriers. SUMMARY: During the period of this review, the importance of several known risk factors was confirmed, whereas the effects of other factors became more clear. Chemoprevention and prophylactic surgery have emerged as preventative options that can reduce the risk of breast and ovarian cancer. PMID- 12544505 TI - Positron emission tomography scanning in gynecologic and breast cancers. AB - PURPOSE OF REVIEW: Positron emission tomography with 2-[fluorine-18]-fluoro-2 deoxy-D-glucose represents a noninvasive functional imaging modality that is based on metabolic characteristics of malignant tumors. The recent findings of this technique in breast cancer, cervical cancer, ovarian cancer, and other gynecologic malignancies are discussed. RECENT FINDINGS: In breast cancer, positron emission tomography with 2-[fluorine-18]-fluoro-2-deoxy-D-glucose is more accurate than conventional methods for the staging of distant metastases, enables early assessment of treatment response in patients undergoing primary chemotherapy. The diagnostic accuracy for axillary lymph node staging depends on the tumor load of the lymph nodes. The sensitivity of this technique in detecting primary breast cancer is limited in small breast lesions and invasive lobular cancer. In cervical cancer it is the most accurate noninvasive method for lymph node staging and it can accurately depict recurrent ovarian cancer in patients with elevated CA125 levels. False negative findings in well differentiated adenocarcinoma and borderline lesions as well as false positive findings in benign conditions limit the role of positron emission tomography scanning for the differential diagnosis of adnex tumors. SUMMARY: Positron emission tomography with 2-[fluorine-18]-fluoro-2-deoxy-D-glucose reveals unique information about tumor metabolism in gynecologic malignancies and breast cancer. This technique is complementary to morphological imaging for primary diagnosis, staging and re staging. It may become the method of choice for the early assessment of treatment response in breast cancer and the detection of recurrent disease in ovarian cancer. This method, however, cannot replace invasive procedures if microscopic disease is of clinical relevance. PMID- 12544506 TI - Array of possibilities. PMID- 12544507 TI - Indexing pharmacogenetic knowledge on the World Wide Web. PMID- 12544508 TI - A microarray minisequencing system for pharmacogenetic profiling of antihypertensive drug response. AB - We aimed to develop a microarray genotyping system for multiplex analysis of a panel of single nucleotide polymorphisms (SNPs) in genes encoding proteins involved in blood pressure regulation, and to apply this system in a pilot study demonstrating its feasibility in the pharmacogenetics of hypertension. A panel of 74 SNPs in 25 genes involved in blood pressure regulation was selected from the SNP databases, and genotyped in DNA samples of 97 hypertensive patients. The patients had been randomized to double-blind treatment with either the angiotensin II type 1 receptor blocker irbesartan or the beta 1-adrenergic receptor blocker atenolol. Genotyping was performed using a microarray based DNA polymerase assisted 'minisequencing' single nucleotide primer extension assay with fluorescence detection. The observed genotypes were related to the blood pressure reduction using stepwise multiple regression analysis. The allele frequencies of the selected SNPs were determined in the Swedish population. The established microarray-based genotyping system was validated and allowed unequivocal multiplex genotyping of the panel of 74 SNPs in every patient. Almost 7200 SNP genotypes were generated in the study. Profiles of four or five SNP genotypes that may be useful as predictors of blood pressure reduction after antihypertensive treatment were identified. Our results highlight the potential of microarray-based technology for SNP genotyping in pharmacogenetics. PMID- 12544509 TI - Natural allelic variants of breast cancer resistance protein (BCRP) and their relationship to BCRP expression in human intestine. AB - The aim of this study was to identify the extent of genetic variability in breast cancer resistance protein (BCRP) in humans. We first analysed the sequence of BCRP cDNA from human livers and from human intestines phenotyped for expression of intestinal BCRP. We then determined the frequency of all known coding single nucleotide polymorphisms (cSNPs) using DNA from individuals representing 11 different ethnic populations. Nine SNPs including four non-synonymous and three synonymous cSNPs and two intronic SNPs were identified. Of the missense mutations, exon 2 SNP (G34A) resulted in a V12M change; exon 5 SNP (C421A) resulted in a Q141K substitution; exon 6 SNP (A616C) resulted in an I206L amino acid substitution; and exon 15 SNP (A1768T) resulted in a N590Y change in the BCRP protein. The two most frequent polymorphisms identified in the human population studied were the G34A and C421A transitions. There was marked variation in BCRP genotypes and allele frequencies in the different populations. BCRP mRNA was phenotyped in human small bowel intestinal samples by real-time polymerase chain reaction and BCRP protein was analysed on immunoblots of tissue from the same individuals. There was a 78-fold variation in expression of BCRP mRNA and significant variation in BCRP protein expression in human intestine. Expression of intestinal BCRP mRNA and protein was not different between persons expressing the common Gln141 allele compared to the Lys141 allele. Thus, common natural allelic variants of BCRP have been identified, and did not influence interindividual variation in expression of BCRP mRNA in human intestine, but remain to be tested for their effect on BCRP function. PMID- 12544510 TI - A functional single-nucleotide polymorphism in the human cytidine deaminase gene contributing to ara-C sensitivity. AB - To test the hypothesis that analyses of drug targets for polymorphism will help to establish gene-based information for the treatment of cancer patients, we investigated the functional single-nucleotide polymorphisms in the human cytidine deaminase (HDCA) gene. The cDNAs from 52 leukaemia/lymphoma samples and 169 control blood samples were direct-sequenced and analysed for the polymorphisms. Three different polymorphisms (A79C, G208A and T435C) were identified in the coding region of the HDCA gene and displayed allelic frequencies of 20.1%, 4.3% and 70.1%, respectively. No association with susceptibility to disease was observed. A novel polymorphism, G208A produced an alanine to threonine substitution (A70T) within the conserved catalytic domain. By introduction of the polymorphic HCDA genes into the yeast CDA-null mutants, the HCDA-70T showed 40% and 32% activity of prototype for cytidine and ara-C substrates, respectively (P < 0.01). The ara-C IC50 value of the yeast transformants carrying HCDA-70T was 757 +/- 33 micromol and was significantly lower (P < 0.01) than that of prototype (941 +/- 58 micromol). This study demonstrated a population characterized with 208A genotype for, which potentially leads one more sensitive to ara-C treatment than prototype. Accumulation of polymorphisms in the genes responsible for drug metabolism and determination of polymorphism-induced biological variations could provide the additional therapeutic strategies in risk-stratified protocols for the treatment of childhood malignancies. PMID- 12544511 TI - Role of CYP2D6 in the stereoselective disposition of venlafaxine in humans. AB - CYP2D6 is involved in the O-demethylation metabolic pathway of venlafaxine in humans. In this study, we investigated whether this isozyme is stereoselective. Plasma samples from seven CYP2D6 extensive metabolizers (EMs) and five CYP2D6 poor metabolizers (PMs), collected during a period without and with coadministration of quinidine, were analysed. Subjects were administered venlafaxine hydrochloride 18.75 mg orally every 12 h for 48 h on two occasions (1 week apart); once alone and once during the concomitant administration of quinidine sulphate every 12 h. Blood and urine samples were collected under steady-state conditions over one dosing interval (12 h). The present results show that, although CYP2D6 catalyses the O-demethylation of both enantiomers of venlafaxine, it displays a marked stereoselectivity towards the (R)-enantiomer. The oral clearance of (R)-venlafaxine was found to be nine-fold higher in EMs compared to PMs [median (range) 173 (29-611) l/h versus 20 (16-24) l/h, P < 0.005], while it was two-fold higher for (S)-venlafaxine [73 (32-130) l/h versus 37 (21-44) l/h, P < 0.05]. In EMs, quinidine decreased (R)- and (S)-venlafaxine oral clearance by 12-fold ( 0.05) and four-fold ( 0.05), respectively. In contrast, quinidine did not have any effects on renal clearance of (R) venlafaxine [4 (2-10) l/h for venlafaxine alone versus 5 (0.6-7) l/h for venlafaxine + quinidine] and of (S)-venlafaxine [4 (1-7) l/h for venlafaxine alone versus 3 (0.4-6) l/h for venlafaxine + quinidine]. The coadministration of quinidine to EMs resulted in an almost complete inhibition of the partial metabolic clearance of (R)-venlafaxine to O-demethylated metabolites [127 (10 493) l/h down to 1 (0.1-3) l/h, 0.05], while a seven-fold reduction was measured for (S)-venlafaxine [47 (14-94) l/h versus 7 (1-19) l/h, 0.05]. In PMs, coadministration of quinidine did not significantly change oral clearance and partial metabolic clearance of (R)- and (S)-venlafaxine to its various metabolites. In contrast, data obtained on the partial metabolic clearance of (R) and (S)-venlafaxine to N-demethylated metabolites, a reaction which is mediated by CYP3A4, suggest a lack of stereoselectivity of this enzyme. PMID- 12544512 TI - Hepatocyte nuclear factor-4 alpha/gamma and hepatocyte nuclear factor-1 alpha as causal factors of interindividual difference in the expression of human dihydrodiol dehydrogenase 4 mRNA in human livers. AB - Human dihydrodiol dehydrogenase (DD) catalyses the oxidation of trans dihydrodiols of polycyclic aromatic hydrocarbons and the reduction of several ketone-containing drugs. About 40-fold interindividual difference in DD activities has been noted. Recently, we found that transcriptional factors, hepatocyte nuclear factor (HNF)-1 alpha, HNF-4 alpha and HNF-4 gamma were essential for the expression of DD4 mRNA, which is a major form of DDs. Thus, to clarify a possible mechanism(s) underlying the interindividual difference in DD activities, we investigated the sequences of genes and the expression levels of mRNA for DD4 and HNFs in human livers. We found no clear relationship between the genotypes of DD4 and HNF genes and the expression levels of DD4 mRNA in the subjects. The expression level of DD4 mRNA significantly correlated with that of HNF-1 alpha, HNF-4 alpha or HNF-4 gamma. These results suggest that the expression level of DD4 mRNA is cooperatively regulated by the amounts of HNF-1 alpha, HNF-4 alpha and HNF-4 gamma. PMID- 12544513 TI - Arylamine N-acetyltransferase (NAT2) genotypes in Africans: the identification of a new allele with nucleotide changes 481C>T and 590G>A. AB - This study was carried out to characterize the distribution of NAT2 allelic variants among a sample of three African populations. We determined the frequencies of major NAT2 allele clusters (NAT2*4, *6, *7 and *14) using PCR/restriction fragment length polymorphism and sequencing techniques. The genotypes predict slow acetylator phenotypes of 49, 38 and 52% among Tanzanians, Venda and Zimbabweans, respectively. The most common genotype was NAT2*4/*5. NAT2* 5 was the most common allele while NAT2* 7 was the least common. A new allele with two base changes occurring together, 481C>T and 590G>A, is reported. The frequency of the occurrence of the combination 481C>T and 590G>A, was found to be 9% (30/326), 7% (14/192) and 8% (18/234) among Zimbabweans, Venda and Tanzanians, respectively. The allele has been named NAT2*6E. Among Africans, the change 481C>T is not only associated with 341C>T (i.e. the NAT2* 5 allele cluster) as in other populations, but also with 590G>A on the same allele. PMID- 12544514 TI - A picture from Philadelphia. PMID- 12544515 TI - Dysplasia of the anal transitional zone after ileal pouch-anal anastomosis: results of prospective evaluation after a minimum of ten years. AB - PURPOSE: Stapling of the ileal pouch-anal anastomosis with preservation of the anal transitional zone remains controversial because of concerns about the potential risk of dysplasia and cancer. The natural history and optimal treatment of anal transitional zone dysplasia ten or more years after surgery are unknown. This study establishes the risk of dysplasia in the anal transitional zone and the outcome of a conservative management policy for anal transitional zone dysplasia, with a minimum of ten years' follow-up after ileal pouch-anal anastomosis. METHODS: A total of 289 patients undergoing anal transitional zone sparing stapled ileal pouch-anal anastomosis for inflammatory bowel disease between 1986 and 1990 were studied. Patients undergoing anal transitional zone sparing ileal pouch-anal anastomosis who were studied with serial anal transitional zone biopsies for at least ten years postoperatively were included (n = 178). Median follow-up was 130 (range, 120-157) months. RESULTS: Anal transitional zone dysplasia developed in 8 patients 4 to 123 (median, 9) months after surgery. There was no association with gender, age, preoperative disease duration, or extent of colitis, but the risk of anal transitional zone dysplasia was significantly associated with cancer or dysplasia as a preoperative diagnosis or in the proctocolectomy specimen. Dysplasia was high grade in two patients and low grade in six. Two patients with low-grade dysplasia on two or more occasions after detection of low-grade dysplasia underwent completion mucosectomy and perineal pouch advancement with neo-ileal pouch-anal anastomosis. One patient with high-grade dysplasia on two occasions was to undergo completion mucosectomy, but this was not technically feasible. Partial mucosectomy with vigorous anal transitional zone biopsy was performed with close postoperative surveillance. Biopsies were negative for dysplasia. The second recently diagnosed patient with high-grade dysplasia underwent examination under anesthesia with negative anal transitional zone biopsies and will be kept under close surveillance. No cancer in the anal transitional zone was found during the study period. The 4 other patients with low-grade dysplasia on 1 or 2 occasions were treated expectantly and have been dysplasia free for a median of 119 (range, 103-133) months. CONCLUSIONS: Anal transitional zone dysplasia after stapled ileal pouch-anal anastomosis is infrequent and is usually self-limiting. Anal transitional zone preservation did not lead to the development of cancer in the anal transitional zone with a minimum of ten years of follow-up. Long-term surveillance is recommended to monitor dysplasia. If repeat biopsy confirms persistent dysplasia, mucosectomy with perineal pouch advancement and neo-ileal pouch-anal anastomosis is recommended. PMID- 12544516 TI - Nocturnal penile tumescence is diminished but not ablated in postproctectomy impotence. AB - PURPOSE: We aimed to assess objectively the integrity of the parasympathetic neural pathway that controls the inflow choke vessels to the corpora cavernosa in a group of male patients with postproctectomy erectile dysfunction. METHODS: The study group was male patients with erectile dysfunction after proctectomy for rectal cancer and inflammatory bowel disease identified by sexual function questionnaire. The group underwent two consecutive nights of home nocturnal penile tumescence monitoring with the Nocturnal Electrobioimpedance Volumetric Assessment device. The control group was also monitored. It comprised preoperative potent patients with rectal cancer and inflammatory bowel disease who had not yet undergone a variety of surgical procedures. Demographics and nocturnal penile tumescence parameters were recorded, including number, duration, and percentage increase in penile volume of tumescent events. RESULTS: Thirty four impotent study group and 28 potent control group patients underwent nocturnal penile tumescence monitoring. The groups were well matched for mean age (difference, 1.4 years; 95 percent confidence interval, -5.8 to 8.6 years) and proportion with rectal cancer (difference, 6 percent; 95 percent confidence interval, -1 to 13 percent). The number of nocturnal penile tumescent events was greater for the potent group than for the control group (mean rank, 40.4 vs. 24.2; P = 0.0004). There was no significant difference between the mean duration (difference, 2.6 minutes; mean rank, 27.9 vs. 34.4; P = 0.16) or the mean penile volume increase (difference, 5.4 percent increase; mean rank, 30.6 vs. 32.6; P = 0.66) for tumescent events between the study and control groups. Mean age was significantly higher in complete than in partial impotence (60.9 vs. 53.1 years; difference, 7.8 years; 95 percent confidence interval, 0.1 to 15.5 years). There was a nonsignificant trend to a lower mean number of tumescence events among sildenafil responders than among nonresponders (3.5 vs. 4.8 events; mean rank, 11.2 vs. 17.3; P = 0.14). CONCLUSION: Nocturnal penile tumescence activity is diminished but not ablated by the trauma of surgical dissection. This suggests that some of the cavernous nerves that govern inflow to the corpora cavernosa are intact after surgery and that the nerve lesion responsible for erectile dysfunction is partial, and it explains why the response to sildenafil in such patients is surprisingly high. PMID- 12544517 TI - Anal canal strictures after ileal pouch-anal anastomosis. AB - PURPOSE: This study was designed to define the different types of strictures, the factors favoring their occurrence, and their treatment after ileal pouch-anal anastomosis. METHODS: Between January 1981 and June 1996, 1,884 ileal pouch-anal anastomoses were constructed at the Mayo Clinic in Rochester, Minnesota. Data were collected prospectively and included age, gender, type of underlying diseases (chronic ulcerative colitis familial adenomatous polyposis), proctologic antecedents, technique of anastomosis, intraoperative difficulties, and postsurgical complications. Strictures were categorized as nonfibrotic and fibrotic on the basis of the presence or absence of a fibrotic segment at the anal canal anastomosis that was responsible for pouch-outlet obstruction requiring at least one dilation. RESULTS: Strictures occurred in 213 patients (11.2 percent; 11 percent for chronic ulcerative colitis and 12 percent for familial adenomatous polyposis; P = not significant). Strictures were nonfibrotic in 86.4 percent of patients and fibrotic in 13.6 percent. A greater number of strictures were observed after a handsewn anastomosis (12 percent) than after a stapled anastomosis (4 percent; P = 0.03). Intraoperative technical difficulties were associated with 13 percent of all strictures regardless of the type of stricture (fibrotic, 7.5 percent; nonfibrotic, 14 percent; P = 0.4). Postoperative complications such as abscess, fistula, and pouch retraction were found in 13 percent of cases and were primarily associated with fibrotic strictures. Treatment included dilation, which was successful in 95 percent of nonfibrotic strictures but in only 45 percent of fibrotic strictures (P = 0.0001). Surgical treatment was required in 25 strictures (12 percent), including excision of the strictured segment with mucosal advancement flap (5 patients), excision of the pouch with permanent ileostomy (9 patients), or redo pouch (3 patients). With one exception, all excised pouches were associated with other perianastomotic complications, such as abscess, fistula, and pouch retraction. The remaining eight patients had other surgical procedures because of abscess (n = 3), division of an obstructive bridge (n = 2), and debridement and curettage of a fistula (n = 3) with dilation for associated strictures. CONCLUSIONS: Strictures were observed in 11.2 percent of the patients in this study. Nonfibrotic strictures responded well to anal dilation, whereas fibrotic strictures were more commonly associated with intraoperative or postoperative complications, often necessitated surgical therapy to salvage pouch function, and were eventually responsible for pouch failure in nine patients. PMID- 12544518 TI - Emergency surgery for colon carcinoma. AB - PURPOSE: Emergency surgery for colon cancer is widely thought to be associated with increased likelihood of surgical morbidity and mortality; however, other coexistent factors such as advanced disease, the age of the patient, and medical comorbid conditions may also influence these outcomes. The primary purpose of this study was to identify the relative risk for surgical morbidity and/or mortality conferred by emergency surgery compared with elective surgery for patients with colon cancer. METHODS: An Institutional Review Board-approved, case control study was performed. During the period from January 1, 1995, to June 30, 2001, a total of 184 primary surgeries for colon cancer were performed. Emergency indications for surgery were defined as peritonitis, intra-abdominal abscess, or complete bowel obstruction at presentation (defined as emesis, distention on examination, and confirmatory plain radiograph films). By this definition, 29 patients (15.7 percent) met the criteria for inclusion. These patients were age and stage matched with 29 patients derived from the remaining 155 patients. Information was collected on surgical morbidity and mortality, length of stay, and survival. RESULTS: Age, medical comorbidities, and stage of disease were well matched between groups. The indications for the 29 emergency surgeries were as follows: 6 for peritonitis, 2 for abscesses, and 21 for complete obstructions. Nine patients did not have their primary tumor removed. Sixteen patients underwent resection and anastomosis; the remaining four patients underwent a Hartmann's procedure. Overall surgical morbidity (64 vs. 24 percent; odds ratio, 5.1; 95 percent confidence interval, 1.7-16) and mortality (34 vs. 7 percent; odds ratio, 7.1; 95 percent confidence interval, 1.4-36.2) were significantly higher for patients undergoing emergency surgery. Among patients surviving surgery, there was no difference in overall survival between patients undergoing emergency compared with elective operation. CONCLUSIONS: Emergency surgery has a strong negative influence (beyond that which is expected based on stage of disease) on immediate surgical morbidity and mortality. The similarity between the two groups in overall survival for patients surviving the perioperative period suggests that the negative impact of emergency surgery is confined to the immediate postoperative period. PMID- 12544519 TI - Indicators for surgical resection and intraoperative radiation therapy for pelvic recurrence of colorectal cancer. AB - PURPOSE: We retrospectively analyzed prognostic factors for surgical resection and intraoperative radiation therapy to identify indicators for this treatment strategy. METHODS: Thirty-nine consecutive patients with locally recurrent colorectal cancer who underwent surgical resection with intraoperative radiation therapy from January 1, 1987, to June 30, 1999, were analyzed. The mean electron energy was 10.5 MeV and the mean intraoperative radiation dose was 22.6 Gy. Kaplan-Meier survival estimates were obtained for the 37 patients who recovered postoperatively. Prognostic factors were analyzed univariately by log-rank test and multivariately by Cox's proportional hazards model. RESULTS: Three-year cumulative survival was 44 percent (standard error = 11) for 26 patients free of unresectable distant metastasis who underwent surgical resection and intraoperative radiation therapy for pelvic recurrence of colorectal cancer, but none of the 11 patients with unresectable distant metastasis survived 3 years. Preoperative prognostic factors which were significant on univariate and multivariate analysis were unresectable distant metastasis (P = 0.001) and elevated preoperative serum CA 19-9 (P = 0.02). Patients with synchronous resection of local recurrence and distant metastasis had a significant survival advantage over those without resection of metastases (P = 0.02). Univariate analysis in a subgroup of 26 patients without unresectable distant metastasis revealed pain (P = 0.0003) to be a useful preoperative prognostic indicator, whereas tumor fixation (P = 0.01) and amount of residual tumor after surgical resection (P = 0.01) were significant intraoperative and postoperative factors, respectively. Fluorouracil-based postoperative systemic chemotherapy produced a significant survival benefit (P = 0.04). CONCLUSIONS: Patients with unresectable distant metastasis are not suitable candidates for surgical resection and intraoperative radiation therapy, whereas those with resectable metastasis are potential candidates. Intraoperative radiation therapy may be less useful for patients with pain, elevated preoperative CA19-9, fixed tumors, or gross residual tumor after surgical resection. Multimodal treatment strategies combining preoperative and/or postoperative external beam radiation therapy and intraoperative radiation therapy with fluorouracil-based systemic chemotherapy are recommended for patients with these indicators. PMID- 12544520 TI - Lymphatic vessels in colorectal cancer and their relation with inflammatory infiltrate. AB - PURPOSE: The aim of this study was to determine why colorectal tumors confined to submucosa rarely metastasize. Under normal conditions, the submucosa contains many large lymphatic vessels with thin walls that would presumably favor the spread of cancer cells through the lymphatic system. METHODS: Specimens of colorectal cancer tissue, the border between tumor and normal tissue, and normal tissue were obtained from patients undergoing radical resection of colorectal cancer. The material was embedded in methacrylate resin for light microscopy and Epon for transmission electron microscopy examination. Light microscopy observations were routinely performed on serial sections. RESULTS: No lymphatic vessels were ever found in the tumor mass. The border area contained peritumoral inflammatory infiltrate of variable thickness. Where submucosal lymphatic vessels came into contact with peritumoral inflammatory infiltrate, they were profoundly altered: their endothelium was fragmented, and their walls were disrupted. These altered lymphatic vessels were almost always accompanied by mast cells, which were observed in the process of degranulating toward the lymphatic endothelium. No such alterations were detected in blood vessels. CONCLUSION: Our results suggest that mast cells, probably influenced by inflammatory infiltrate and/or colorectal cancer cells, destroy lymphatic vessels, which prevents cancer cells from spreading through the lymphatic system. PMID- 12544521 TI - Long-term results of polyp clearance by intraoperative enteroscopy in the Peutz Jeghers syndrome. AB - PURPOSE: Enteroscopy during laparotomy for Peutz-Jeghers syndrome was introduced in our unit in 1987. Its aim is to achieve more complete polyp clearance and thereby reduce the number of subsequent laparotomies for small intestinal polyps. METHODS: All patients with Peutz-Jeghers syndrome who had undergone intraoperative enteroscopy since its introduction into our unit in 1987 were identified. The numbers of polyps identified by palpation and transillumination and by enteroscopy were recorded. The timing of, indications for, and findings of all subsequent laparotomies were analyzed and compared with data from our unit before the introduction of intraoperative enteroscopy. RESULTS: Twenty-five patients (14 females) were studied. Enteroscopy identified 350 (median 12, range 0-35) polyps not detected by palpation and transillumination. All impalpable polyps were removed endoscopically by snare or biopsy. The median follow-up was 53 (interquartile range, 13-133) months. Six patients have had an additional laparotomy (1 urgent relaparotomy for small-bowel perforation after endoscopic polypectomy, 4 polypectomies, and 1 adhesion obstruction). No patient has required operative polypectomy within 4 years of polyp clearance by intraoperative enteroscopy, compared with registry data of 4 (17 percent) of 23 patients who had more than 1 laparotomy within 1 year. CONCLUSION: Intraoperative enteroscopy for Peutz-Jeghers syndrome improves polyp clearance without the need for additional enterotomies and may help to reduce the frequency of laparotomies. PMID- 12544522 TI - Cluster randomization trial of sequence mass screening for colorectal cancer. AB - PURPOSE: Colorectal cancer is a major cause of death worldwide. To reduce the incidence and mortality from rectal cancer, an individual quantitative risk assessment model (hereafter referred to as the Attributive Degree Value) and reverse passive hemagglutination fecal occult blood test were used in a randomized, controlled, population-based trial that was conducted in Jiashan County, People's Republic of China. METHODS: All residents of Jiashan County aged 30 years or older were enrolled in the study, and 21 townships in the county were randomized to either a screening (n = 10 townships) or control (n = 11 townships) group. Participants in the screened group submitted a one-article-per-slide stool sample and completed a structured risk-assessment questionnaire from which their attributive degree value was computed. According to study protocol, 4,299 participants were defined as high risk and underwent diagnostic evaluation with 60-cm flexible sigmoidoscopy and, in some cases, an additional screening with colonoscopy. RESULTS: From 1989 to 1996, cumulative mortality from colon cancer was 90 (95 percent confidence interval, 83-97) per 100,000 in the screened group and 83 (95 percent confidence interval, 76-90) per 100,000 in the control group (log-rank P = 1.49, P = 0.222). Mortality from rectal cancer during this time was 110 (95 percent confidence interval, 102-118) per 100,000 in the screened group, which differed significantly from the control group mortality rate of 161 (95 percent confidence interval, 152-170) per 100,000 (log-rank P = 0.003). CONCLUSION: Mass screening with a reverse passive hemagglutination fecal occult blood test along with an individual attributive degree value score was effective in reducing mortality from rectal cancer but not in reducing mortality from colon cancer or the incidence of colorectal cancer. PMID- 12544523 TI - Combined-modality therapy in locally advanced primary rectal cancer. AB - PURPOSE: Patients with unresectable, locally advanced rectal cancer are reported to have a dismal prognosis. The aim of this study was to analyze the effect of combined-modality therapy on clinical outcome. METHODS: From March 1990 to December 1997, 43 patients (28 males; median age, 62 years; median follow-up, 74 months) with locally advanced (T4 and/or N3) nonmetastatic rectal cancer received external-beam radiation (23.6 plus 23.6 Gy (split course), 8 patients; 45 Gy, 35 patients) plus 5-fluorouracil (96-hour continuous infusion, Days 1-4, at 1,000 mg/m(2)/day) and mitomycin C (10 mg/m, intravenous bolus, Day 1). Concomitant chemotherapy was repeated at the beginning of the second course (split-course group) or in the last week of radiotherapy (continuous-course group). After 6 to 8 weeks, patients were evaluated for surgical resection and intraoperative radiation therapy (10 to 15 Gy). Thereafter, adjuvant chemotherapy (5 fluorouracil plus leucovorin, 6-9 courses) was prescribed. RESULTS: During chemoradiation, 5 patients (11.6 percent) developed Grade 3 to 4 hematologic toxicity. After chemoradiation, 29 patients (67.4 percent) had an objective clinical response (complete response, 2.3 percent; partial response, 65.1 percent). Thirty-eight patients underwent radical surgery (anterior resection, 24 patients; abdominoperineal resection, 14 patients; intraoperative radiation therapy boost on the tumor bed, 19 patients), and 2 patients had partial tumor resection. No perioperative deaths occurred in the patient group. Five-year survival and local control rates were 59.9 and 69.1 percent, respectively. Distant metastasis occurred in 44.2 percent of patients. Statistically significant relationships between intraoperative radiation therapy and local control (P = 0.0104), radical surgery and survival (P = 0.0120), and adjuvant chemotherapy and disease-free survival (P = 0.0112) were observed. CONCLUSIONS: Our data suggest that combined-modality therapy was relatively well tolerated and resulted in good local control and survival. With regard to the impact of surgical resection on survival, additional studies aimed at improving the local response rate are necessary, whereas the positive impact of intraoperative radiotherapy on local control appears to justify the inclusion of this therapeutic modality in prospective multi-institutional trials. PMID- 12544524 TI - Scintigraphic assessment of retrograde colonic washout in fecal incontinence and constipation. AB - PURPOSE: This study aimed to evaluate the colorectal luminal transport obtained by retrograde colonic washout with a new scintigraphic technique. METHODS: Nineteen patients (5 with spinal cord lesion, 6 with idiopathic fecal incontinence, and 8 with idiopathic constipation) treated with retrograde colonic washout took indium-111-labeled polystyrene pellets to label the bowel contents. Technetium-99m-diethylene-triamine-pentaacetic acid was mixed with the irrigation fluid to assess its extent within the colorectum. Scintigraphy was performed before and after a standardized washout procedure. The colorectum was divided into four segments: the cecum and ascending colon, the transverse colon, the descending colon, and the rectosigmoid. Assuming ordered evacuation of the colorectum, the contribution of each colonic segment to the total evacuation was expressed in percent of the original segmental counts. The contributions of each segment were summed to reach a total defecation score (range, 0-400), and directional segmental transports were estimated. RESULTS: The defecation score in patients with idiopathic constipation (median, 59; range, 21-130) differed significantly (P < 0.05) from the scores in those with spinal cord lesions (median, 204; range, 108-323) and idiopathic fecal incontinence (median, 188; range, 155-234). Thus, patients with spinal cord lesion or idiopathic fecal incontinence were able to empty most of the rectosigmoid and most of the descending colon, but those with idiopathic constipation could only empty 59 percent of the rectosigmoid. The irrigation fluid on average reached a point just beyond the right colic flexure that correlated with the defecation score (r(2) = 0.58, P < 0.001). CONCLUSION: The effect of retrograde colonic washout was significantly better in spinal cord lesion and idiopathic fecal incontinence than in idiopathic constipation, and its effect correlated with the extent to which the irrigation fluid had entered the colorectum. PMID- 12544525 TI - Linear pressure profiles and symmetric findings in the stimulated gracilis muscle. AB - PURPOSE: This is the first report of linear pressure profiles and symmetric findings in preoperative and postoperative (after gracilis muscle retraining) patients. Length of the anal canal and symmetry have been well documented in the literature as significant parameters in the maintenance of anal incontinence. Significant improvement of these parameters should play a critical role in the functional outcome of the stimulated gracilis. METHODS: Six patients underwent preoperative and postoperative anorectal manometry using an eight-channel radial catheter. Pressures were collected at 8 Hz using an automated continuous pullout technique (1 mm/sec). Preoperative and posttraining resting and squeeze pressures, pressure volumes, anal canal length, and linear and cross-sectional images were compared. RESULTS: Resting pressures significantly improved from 13.7 mmHg to 26.8 mmHg (P < 0.02). The squeeze pressure significantly improved from 28.67 mmHg to 62.9 mmHg (P < 0.02). The pressure volume increased from 10,429 mmHg to 26,162 mmHg. Anal canal length increased from an abnormal length of 2.95 cm to normal length of 3.55 cm (P < 0.04). The percentage of sphincter asymmetry decreased from 28.6 percent to 23.5 percent (P < 0.01). The most striking observation was the impact the gracilis muscle had on correcting the geographic cross sections as it wrapped more than 360 degrees from the proximal to distal anal canal. Patients were significantly improved, with four of the six patients completely continent to solid stool. Two of the six patients had minor episodes of seepage. Preoperatively, all patients were grossly incontinent to solid stool. CONCLUSIONS: Significant improvement in resting pressure, pressure volume, anal canal length, and cross-sectional symmetry has led to a significant improvement in anal incontinence after dynamic myoplasty. PMID- 12544526 TI - Reduced expression of serotonin receptor(s) in the left colon of patients with colonic inertia. AB - PURPOSE: Serotonin regulates colonic motility receptors expressed on neural fibers and smooth muscle. Colonic inertia is characterized by delayed colonic transit. Abnormalities in serotonin receptor protein, as judged by immunoreactivity levels, could contribute to the origin of colonic inertia. The aim of this study was to investigate the expression of serotonin receptor(s) immunoreactivity in the left colon of patients with colonic inertia compared with controls. METHODS: Sixteen patients who underwent subtotal colectomy for colonic inertia were assessed. Colonic transit time was measured with the radiopaque marker technique and presented as the number of retained markers in the colon on Day 5. The control group consisted of 18 patients who underwent left hemicolectomy for colonic carcinoma; histologically normal tissues from the left colon were used. Immunohistochemical staining for serotonin receptor was performed with a rabbit anti-idiotypic antibody. The average positive area (square pixels) in the mucosa, muscularis mucosa, submucosa, and circular and longitudinal muscles per microscopic field (63x) was calculated based on measurement of the positively stained area in 20 randomly chosen microscopic fields in each related structure. The Scion Image computer analysis system was used. RESULTS: Serotonin receptor(s) immunoreactivity was mainly detected in the muscular mucosa, circular muscles, and longitudinal muscles and rarely in the mucosa and submucosa. In muscularis mucosa and circular muscle, the positive areas were significantly less in the colonic inertia group than in controls (muscularis mucosa: 29.1 +/- 10.8 vs 109.7 +/- 28.2, P < 0.05; circular muscle: 25.6 +/- 6.2 vs 90.2 +/- 19.1, P < 0.01). There were significantly positive correlations in the control group in serotonin receptor(s) immunoreactivity levels between circular muscle and longitudinal muscle (r = 0.54, P < 0.05) and between muscular mucosa and longitudinal muscle (r = 0.57, P < 0.05) but not in colonic inertia patients. In addition, the positive areas in the circular muscle were positively correlated to the colonic transit time (Spearman's rank correlation, 0.83; P < 0.01). CONCLUSION: In colonic inertia patients, the serotonin receptor(s) immunoreactivity level is lower in muscular mucosa and circular muscle. The absence of a correlation of serotonin receptor(s) immunoreactivity in the muscular mucosa and muscularis propria in the patient group implies that an uncoordinated expression of serotonin receptors may also contribute to colonic inertia. However, the positive correlation between serotonin receptor(s) immunoreactivity levels in the circular muscle and the transit time observed in colonic inertia patients suggests a decrease in stimulatory subtypes and at the same time an increase in inhibitory subtypes of serotonin receptors in this tissue. PMID- 12544527 TI - Clinical experience of sutureless closed hemorrhoidectomy with LigaSure. AB - PURPOSE: The purpose of this study was to evaluate the LigaSure vessel sealing system as an alternative to closed hemorrhoidectomy. METHODS: Sixty-one patients with Grade 3 or 4 symptomatic hemorrhoids were prospectively randomly assigned to undergo hemorrhoidectomy with the LigaSure vessel sealing system or hemorrhoidectomy using the conventional Ferguson procedure. We determined the operation time, postoperative pain, amount of time taken off from work, and complications associated with both techniques. RESULTS: Mean operative time for the LigaSure hemorrhoidectomy was 15 +/- 5.4 minutes and for the Ferguson operation, 21.2 +/- 8.2 minutes. The difference was significant (P < 0.01). There was also a significant decrease in pain measurements reported on postoperative Days 1 and 2 (P < 0.05) in the LigaSure group. The incidence of postoperative wound swelling and complications were similar between two groups. There was no difference in the period of time off from work between patient groups. CONCLUSION: This study confirms that LigaSure system can achieve a radical ablation of hemorrhoids, reduce operative time, and result in less postoperative pain on postoperative Days 1 and 2. PMID- 12544528 TI - Day-case stapled (circular) vs. diathermy hemorrhoidectomy: a randomized, controlled trial evaluating surgical and functional outcome. AB - PURPOSE: Stapled hemorrhoidectomy may be associated with less pain and faster recovery than conventional hemorrhoidectomy for prolapsing hemorrhoids. Therefore, the outcome of stapled hemorrhoidectomy was compared with that of diathermy hemorrhoidectomy in a randomized, controlled trial. METHODS: Sixty patients with third-degree hemorrhoids were randomly assigned to stapled hemorrhoidectomy (n = 30) or to diathermy hemorrhoidectomy in a day-case setting. Visual analog scale was used for postoperative pain scoring. Surgical and functional outcome was assessed at six weeks and one year after surgery. RESULTS: Operation time was a median of 21 (range, 11-59) minutes in the stapled group. 22 (range, 14-40) minutes in the diathermy group. Day-case surgery was successful in 24 patients (80 percent) in the stapled group vs. 29 patients (97 percent) in the diathermy group. Average pain in the stapled group was significantly lower than in the diathermy group (median, 1.8 (0.1-4.8) vs. 4.3 (1.4-6.2), 95 percent confidence interval difference medians, 1.15-3.85, P = 0.0002, Mann-Whitney U test) as was the average pain expected by the patients (median -2.7 (-0.15-0.8) vs. 0.006 (-4.05-0.5) respectively, 95 percent confidence interval difference medians, 0.5-3.55, P = 0.0018, Mann-Whitney U test). Postoperative morbidity and time off work were not significantly different between the diathermy and stapled groups. Seven treatment failures in the stapled group and one in the diathermy group necessitated other treatments at a later date. Patient satisfaction scores in the stapled and diathermy group were similar. Symptoms attributed to difficult rectal evacuation decreased significantly after surgery. CONCLUSIONS: Stapled hemorrhoidectomy is a significantly less painful operation than diathermy hemorrhoidectomy, but does not seem to offer significant advantages in terms of hospital stay or symptom control in the long term. Hemorrhoidectomy may improve symptoms of difficult rectal evacuation. PMID- 12544529 TI - Platelet-activating factor, a critical mediator in the pathogenesis of dextran sulfate sodium-induced colitis in rats. AB - PURPOSE: Disorder of mucosal immunity based on an imbalance between proinflammatory and anti-inflammatory cytokines is believed to be a major factor in the pathogenesis of ulcerative colitis. Platelet-activating factor potentially stimulates the production of proinflammatory cytokines and recruits inflammatory cells. The aim of this study was to determine whether and to what extent platelet activating factor plays a role in the pathogenesis of ulcerative colitis. METHODS: Using dextran sulfate sodium-induced colitis in rats as a model of ulcerative colitis, we analyzed the composition of cellular infiltrates and the local tissue expression of messenger ribonucleic acid for cytokine-induced neutrophil chemoattractant and tumor necrosis factor-alpha. To directly assess the impact of platelet-activating factor on the development of colitis, we also determined the efficacy of a specific platelet-activating factor receptor antagonist for preventing dextran sulfate sodium-induced colitis. RESULTS: The activity of colitis was well correlated with the upregulation of cytokine-induced neutrophil chemoattractant and tumor necrosis factor-alpha messenger ribonucleic acid in local tissues and infiltration of cytokine-induced neutrophil chemoattractant-positive neutrophils and ED1-positive macrophages. The platelet activating factor receptor antagonist effectively ameliorated colitis, along with causing a decrease in the tissue cytokine-induced neutrophil chemoattractant messenger ribonucleic acid level and a decline in neutrophil and macrophage infiltration. However, the antagonist did not alter tissue levels of tumor necrosis factor-alpha messenger ribonucleic acid. CONCLUSION: Platelet-activating factor plays a critical role in the pathogenesis of dextran sulfate sodium induced colitis through recruitment of cytokine-induced neutrophil chemoattractant-positive neutrophils and macrophages and/or stimulation of cytokine-induced neutrophil chemoattractant release from activated neutrophils. The tissue level of tumor necrosis factor-alpha messenger ribonucleic acid does not closely reflect the activity of colitis. PMID- 12544530 TI - Turnbull-Blowhole colostomy for toxic ulcerative colitis in pregnancy: report of two cases. AB - PURPOSE: Surgical intervention for toxic ulcerative colitis in pregnancy is associated with a high morbidity and mortality for both mother and her fetus. A limited surgical procedure may be desirable to reduce the risk of maternal and fetal mortality. We describe the use of "blowhole" colostomy and loop ileostomy (Turnbull procedure) in the management of toxic dilation of the colon complicating ulcerative colitis in pregnancy. METHODS: The medical records of two pregnant females with toxic ulcerative colitis were reviewed. The presentations, management, and outcomes of these patients and their neonates are described. RESULTS: The Turnbull-Blowhole colostomy and ileostomy was used in the acute management of toxic ulcerative colitis in two pregnant females. Both mothers and neonates had successful outcomes with resolution of the acute process. Completion proctocolectomy and ileal pouch-anal anastomosis was later performed in both patients. CONCLUSIONS: The colonic decompression and fecal diversion procedure provided minimal operative trauma to the mother and fetus and resulted in successful outcome in the management of toxic ulcerative colitis in pregnancy. We believe the procedure is safe, effective, and relatively simple, which may reduce the morbidity and mortality of toxic colitis in pregnancy. PMID- 12544531 TI - Rectal perforation: a life-threatening complication of stapled hemorrhoidectomy: report of a case. AB - Stapled hemorrhoidectomy is considered to be safe and carries advantages. We describe a patient with rectal perforation and fecal peritonitis after stapled hemorrhoidectomy. We suggest that it should be performed by experienced colorectal surgeons who are familiar with the technique and aware of possible complications. PMID- 12544532 TI - Behcet's ileocolitis: successful treatment with tumor necrosis factor-alpha antibody (infliximab) therapy: report of a case. AB - Behcet's disease is a chronic inflammatory disease characterized by recurrent oral aphthae and systemic manifestations. Gastrointestinal involvement is rare. We report a case of ileocolitis secondary to Behcet's successfully treated with tumor necrosis factor-alpha antibody (infliximab) therapy. To our knowledge this is the second reported case of Behcet's ileocolitis successfully treated with anti-tumor necrosis factor therapy. PMID- 12544533 TI - Revising the troublesome stoma: combined abdominal wall recontouring and revision of stomas. AB - PURPOSE: Despite preoperative siting and maturation of stomas, some patients may have poor stoma function because of redundant pannus, scars, creases, and parastomal or incisional hernias. In these patients, a combined abdominoplasty and stoma revision may be helpful. The object of this manuscript is to report our preliminary results. METHODS: Eight patients (mean age, 48 years; female/male ratio, 7/1) undergoing this procedure are reported. Five patients had inflammatory bowel disease, two had malignancies, and one had multiple sclerosis with bladder and bowel involvement. Patients were contacted at two months to two years follow-up to assess functional outcome and satisfaction with the procedure. RESULTS: Indications for surgery were difficulty maintaining an appliance (n = 7), frequent stool leakage (n = 6), and skin irritation (n = 4). Multiple surgeries (n = 4), skin creases (n = 3), scarring (n = 2), large weight loss (n = 4), and hernias (n = 5) were contributing factors. At surgery five patients had hernias repaired (3 parastomal, 2 incisional), three had stomas resited, and three underwent resections for Crohn's disease. One patient developed a seroma postoperatively. At follow-up six patients were able to maintain an appliance for at least four days, whereas one changed the appliance every three days because of personal preference. None experience stool leakage. All reported a dramatic improvement in body image. CONCLUSION: Combined stoma revision and abdominoplasty can be performed safely and leads to improved functional results and outcome. PMID- 12544534 TI - A new model of strictureplasty for multiple and long stenoses in Crohn's ileitis: side-to-side diseased to disease-free anastomosis. AB - Conservative surgery has become the treatment of choice for diffuse jejunoileal Crohn's Disease. Previous research described a conservative approach both for multiple strictures located in close proximity to each other and for long stenoses. The side-to-side enteroenteric anastomosis has gained popularity and has lead to nonresectional surgery even for those patients who, until now, were considered suitable only for resection. This technique however, presents in our hands some disadvantages represented mainly by early restenosis at the two edges of the strictureplasty, probably caused by the sutures between the diseased and the thickened part of the bowel. We propose a new technique called "side-to-side diseased to disease-free anastomosis," which consists of dividing the bowel and the mesentery at the beginning of the stenoses and suturing the disease-free bowel above the stenoses to the diseased bowel. This procedure could avoid early restenosis at the two ends of the strictureplasty. Moreover, it is faster and safer to perform because the knots of the sutures are tied to the normal bowel with less risk of bleeding. PMID- 12544535 TI - PPH stapled hemorrhoidectomy--a cautionary note. PMID- 12544536 TI - Grading systems for cost-effectiveness studies: is the whole greater than the sum of the parts? PMID- 12544537 TI - Risk assessment and adjustment: adjusting for sick patients or a sick system? PMID- 12544538 TI - Forecasting the nursing home population. AB - OBJECTIVE: To forecast growth in the US nursing home population, as a function of trends in disability and marriage. METHODS: Nursing home residence is modeled as a function of disability status, marital status, and other demographic covariates. Our predictions for nursing home residence are built upon joint forecasts of marriage and disability. We use data from the 1992 to 1996 Medicare Current Beneficiary Surveys, which are individual-level data sets designed to be representative of the US population older than the age of 65. RESULTS: Today's young cohorts will have higher rates and levels of institutionalization than their older counterparts. This will reverse several decades of decline in rates of disability and institutionalization. The nursing home population is likely to be 10-25% higher than would be suggested by a simple extrapolation of past declines in disability. CONCLUSIONS: In recent years, the rate of institutionalization among the elderly has been falling. It is predicted that this trend will reverse itself within the next decade, and that we will see substantial increases in the incidence of institutionalization among the elderly. This result is generated by our prediction of rising disability among the younger cohorts that are beginning to approach old age. PMID- 12544539 TI - Forecasting the nursing home population. PMID- 12544540 TI - Forecasting health service needs for older adults: some sun, some clouds. PMID- 12544541 TI - A response to the points by Manton and Williamson. PMID- 12544542 TI - Development and validation of a grading system for the quality of cost effectiveness studies. AB - PURPOSE: To provide a practical quantitative tool for appraising the quality of cost-effectiveness (CE) studies. METHODS: A committee comprising [corrected] of health economists selected a set of criteria for the instrument from an item pool. Data collected with a conjoint analysis survey on 120 international health economists were used to estimate weights for each criterion with a random effects regression model. To validate the grading system, a survey was sent to 60 individuals with health economics expertise. Participants first rated the quality of three CE studies on a visual analogue scale, and then evaluated each study using the grading system. Spearman rho and Wilcoxon tests were used to detect convergent validity and analysis of covariance (ANCOVA) for discriminant validity. Agreement between the global rating by experts and the grading system was also examined. RESULTS: Sixteen criteria were selected. Their coefficient estimates ranged from 1.2 to 8.9, with a sum of 93.5 on a 100-point scale. The only insignificant criterion was "use of subgroup analyses." Both convergent validity and discriminant validity of the grading system were shown by the results of the Spearman rho (correlation coefficient = 0.78, P < 0.0001), Wilcoxon test (P = 0.53), and ANCOVA (F(3,146) = 5.97, p = 0.001). The grading system had good agreement with global rating by experts. CONCLUSIONS: The instrument appears to be simple, internally consistent, and valid for measuring the perceived quality of CE studies. Applicability for use in clinical and resource allocation decision-making deserves further study. PMID- 12544543 TI - Development of a model for case-mix adjustment of pressure ulcer prevalence rates. AB - BACKGROUND: Acute care hospitals participating in the Dutch national pressure ulcer prevalence survey use the results of this survey to compare their outcomes and assess their quality of care regarding pressure ulcer prevention. The development of a model for case-mix adjustment is essential for the use of these prevalence rates as an outcome measure. OBJECTIVE: The development of a valid model for case-mix adjustment to compare the prevalence rates in the acute care hospitals that participated in the 1998 Dutch pressure ulcer prevalence survey, for the purpose of performance comparisons among the hospitals. DESIGN: Cross sectional design. SUBJECTS: Subjects were patients residing in the 43 acute care hospitals that participated in the national pressure ulcer prevalence survey on May 26, 1998. MEASURES: The study examined the validity of a model for case-mix adjustment of pressure ulcer prevalence rates and compared hospitals to evaluate the impact of adjusted prevalence rates on their performance. RESULTS: A logistic model was developed for case-mix adjustment, using age, malnutrition, incontinence, activity, mobility, sensory perception, friction and shear, and ward specialty. This model was found to have content, construct, and internal validity. Case-mix adjustment influenced the hospitals' performance. CONCLUSION: The data of the national pressure ulcer prevalence survey can be used to develop a valid model for case-mix adjustment. Conclusions about the quality of care were influenced by the use of case-mix adjusted outcomes as a measure of this quality. PMID- 12544544 TI - The best of both worlds? Potential of hybrid prospective/concurrent risk adjustment. AB - BACKGROUND: There remains considerable uncertainty about whether prospective or concurrent risk adjustment (RA) is preferable. Although concurrent models have better predictive power than prospective models, the large payments associated with concurrent RA create incentives for fraudulent coding. A hybrid strategy--in which prospective payments were used for patients with low expected costs and concurrent payments were available upon the diagnosis of a small number of common, expensive conditions--might improve predictive performance while requiring less auditing than fully concurrent RA. In addition, within-condition RA (using clinical data) for the selected conditions could further improve predictive power. OBJECTIVES: To assess how such a hybrid strategy might perform, focusing on a small number of chronic, expensive conditions that are verifiable (hence auditable). SUBJECTS AND MEASURES: All patients from seven health plans who had two complete years of utilization data were considered. RA models were estimated among patients younger than 65 (n = 319,209) using the Hierarchical Coexisting Conditions (HCC) model with and without stratification of the sample based on the presence of one or more of 100 verifiable, expensive, predictive conditions (VEP100). R2 and predictive ratios were calculated for each model studied. RESULTS: Patients with a VEP100 condition (9.3% of the population) accounted for 84.3% of the variation in cost. R2 was 0.08 using a prospective HCC model on the entire population, but increased to 0.26 for a hybrid using prospective HCCs on the 90.7% of the sample without a VEP100 condition and a simple concurrent model consisting of dummy variables for each of the VEP100 conditions. CONCLUSION: Combined with targeted auditing, a hybrid approach to RA could improve our ability to match payments to costs. However, because this would require additional, costly data collection, more research is needed to determine whether this benefit justifies the data collection and auditing burden. PMID- 12544545 TI - Burden of illness score for elderly persons: risk adjustment incorporating the cumulative impact of diseases, physiologic abnormalities, and functional impairments. AB - BACKGROUND/OBJECTIVES: To develop and validate a new risk adjustment index-the Burden of Illness Score for Elderly Persons (BISEP)-which integrates multiple domains, including diseases, physiologic abnormalities, and functional impairments. RESEARCH DESIGN SUBJECTS: The index was developed in a prospective cohort of 525 patients aged > or = 70 years from the medicine service of a university hospital. The index was validated in a cohort of 1246 patients aged > or = 65 years from 27 hospitals. The outcome was 1-year mortality. RESULTS: Five risk factors were selected from diagnosis, laboratory, and functional status axes: high-risk diagnoses, albumin < or = 3.5 mg/dL, creatinine >1.5 mg/dL, dementia, and walking impairment. The BISEP score (range 0-7) created four groups of increasing risk: group I (score 0-1), group II (2), group III (3), and group IV (> or = 4). In the development cohort, where overall mortality was 154/525 (29%), 1-year mortality rates increased significantly across each risk group, from 8% to 24%, 51%, and 74%, in groups I to IV respectively (chi(2) trend, = 0.001)--an overall 17-fold increased risk by hazard ratio. The c-statistic for the final model was 0.83. Corresponding rates in the validation cohort, where overall mortality was 488/1246 (39%), were 5%, 17%, 33%, and 61% in groups I to IV, respectively (chi(2) trend, = 0.001)-an overall 18-fold increased risk by hazard ratio. The c-statistic for the final model was 0.77. In each cohort, sequential addition of variables from different sources (eg, administrative, laboratory, and chart) substantially improved model fit and predictive accuracy. BISEP had significantly superior mortality prediction compared with five widely used measures. CONCLUSIONS: BISEP provides a useful new risk adjustment system for hospitalized older persons. Although index performance using different data sources has been evaluated, the full BISEP model, incorporating disease, laboratory, and functional impairment information, demonstrates the best performance. PMID- 12544546 TI - Risk adjustment using automated ambulatory pharmacy data: the RxRisk model. AB - OBJECTIVES: Develop and estimate the RxRisk model, a risk assessment instrument that uses automated ambulatory pharmacy data to identify chronic conditions and predict future health care cost. The RxRisk model's performance in predicting cost is compared with a demographic-only model, the Ambulatory Clinical Groups (ACG), and Hierarchical Coexisting Conditions (HCC) ICD-9-CM diagnosis-based risk assessment instruments. Each model's power to forecast health care resource use is assessed. DATA SOURCES: Health services utilization and cost data for approximately 1.5 million individuals enrolled in five mixed-model Health Maintenance Organizations (HMOs) from different regions in the United States. STUDY DESIGN: Retrospective cohort study using automated managed care data. SUBJECTS All persons enrolled during 1995 and 1996 in Group Health Cooperative of Puget Sound, HealthPartners of Minnesota and the Colorado, Ohio and Northeast Regions of Kaiser-Permanente. MEASURES RxRisk, an algorithm that classifies prescription drug fills into chronic disease classes for adults and children. RESULTS: HCCs produce the most accurate forecasts of total costs than either RxRisk or ACGs but RxRisk performs similarly to ACGs. Using the R(2) criteria HCCs explain 15.4% of the prospective variance in cost, whereas RxRisk explains 8.7% and ACGs explain 10.2%. However, for key segments of the cost distribution the differences in forecasting power among HCCs, RxRisk, and ACGs are less obvious, with all three models generating similar predictions for the middle 60% of the cost distribution. CONCLUSIONS: HCCs produce more accurate forecasts of total cost, but the pharmacy-based RxRisk is an alternative risk assessment instrument to several diagnostic based models and depending on the nature of the application may be a more appropriate option for medical risk analysis. PMID- 12544547 TI - An estimation of costs of a pediatric telemedicine practice in public schools. AB - BACKGROUND: Costs per consult are a vital consideration in determining the viability of a telemedicine practice. This study estimated total, average, and marginal cost curves for an ambulatory pediatric telemedicine practice. OBJECTIVE: The objective of this study was to estimate the costs of providing 386 telemedicine clinical consults at school clinics during one school year (September, 1999 to May, 2000). Cost curves were used to estimate costs of clinics with varying numbers of consults. RESEARCH DESIGN: This analysis used cost data from school clinics and a university medical center, which provided physician inputs. Standard cost-accounting procedures were used. SUBJECTS: Cost analyses were conducted using 10 school clinics. Three hundred eighty-six consults were completed with 286 children. Seventy-one percent of the children were black or Hispanic. RESULTS: The average costs of a telemedicine consult ranged from a high of $7328.17 when only one consult was done to a low of $173.13 when 129 consults were completed. At 165 consults, extrapolations from the cost curves suggested that the average costs of a telemedicine consult and a medical center pediatric ambulatory consult were approximately equal, a little more than $153. At 200 consults, telemedicine was estimated to be equal to or less costly than conventional ambulatory care visits by some 9.5%. CONCLUSIONS: Point and CI estimates of average costs suggest that telemedicine is cost competitive with conventional clinics when 200 or more consults are provided. The limited evidence available suggests that telemedicine consults also may be quality competitive. PMID- 12544548 TI - Measuring chronic patients' feelings of being a burden to their caregivers: development and preliminary validation of a scale. AB - BACKGROUND: Burden on caregivers has been extensively studied, but the patient's own feelings of being a burden have rarely been examined. Such feelings may lead to distress and can complicate relations with the caregiver. This report describes the development and preliminary validation of a scale to measure patient-perceived burden. MATERIALS AND METHODS: A conceptual framework and scale items were derived from previous literature and from qualitative interviews with patients and health professionals. Following content validity and item clarity analyses, a 25-item scale was developed. This was then administered in a construct validation to 100 outpatients undergoing hemodialysis. RESULTS: Discriminant validity coefficients showed that burden scores were independent of age, education, and time on dialysis. Convergent analyses showed modest correlations with the number of comorbid conditions (r = 0.20, P < 0.04), functional status (r = 0.26, P < 0.01), physical function (r = 0.30, P < 0.01), and mental health (r = 0.39, P < 0.01). Alpha internal consistency was 0.92 and factor analyses revealed a single main factor. A 10-item abbreviation (alpha = 0.85) is therefore proposed. CONCLUSION: The self-perceived burden scale shows promise as a measure to identify patients in emotional distress due to feelings of being a burden on others, and as an outcome measure in intervention studies. PMID- 12544549 TI - The paradox of dental need in a population-based study of dentate adults. AB - BACKGROUND: Studies typically find that predisposing and enabling factors strongly predict dental utilization, but that need factors do not. However, few longitudinal studies have been conducted, and few have comprehensively measured dental need. OBJECTIVES: To describe the paradox of dental need, and to test three hypotheses regarding need and dental care use. MATERIALS AND METHODS: An observational study that included 873 persons who participated for interview and clinical examination at baseline and 24 months, with 6-month telephone interviews in between. RESULTS: Persons who entered the dental care system during follow-up were actually in better dental health than those who did not. The ability of need factors to predict dental care use, and in which direction, varied with how dental need and the dental care use outcome were measured (eg, care of any type, problem-related care, to receive a dental cleaning, to get a dental checkup). CONCLUSIONS: A substantial number of dental problems remained or developed among the population that did not enter the dental care system. The paradox of dental need has three components: (1) need predicts dental care use but is dependent upon how need is measured; (2) however, persons with a higher probability of new dental problems are actually less likely to seek dental care; and (3) self reported disease and oral pain are associated with a higher likelihood of seeking care, whereas clinically-determined need, such as chewing difficulty, lower self rating, and satisfaction with oral health, are actually associated with a lower likelihood, the former direction being the predominant and expected direction. PMID- 12544550 TI - Lowering generic drug prices: less regulation equals more competition. AB - BACKGROUND: In Ontario, Canada, the 70/90 regulations were instituted in May 1993 to establish provincial government procurement prices for generic drugs. Accordingly, the first generic entrant's price could not exceed 70% of the incumbent's branded price. Subsequent entrants' prices could not exceed 90% of the first entrant's price. OBJECTIVE: These regulations' impact on generic market competitiveness are evaluated. DESIGN AND METHODS: Data on 518 drugs spanning nine therapeutic classifications were collected for the period of 04/01/1987 to 12/31/1998 from Ontario Drug Benefit formulary and IMS Canada. The period 04/01/1987 to 04/30/1993 was defined as the before period (BP) and 05/01/1993 to 12/31/1998 was the after period (AP). We compared the price ratio (P = P(G)/P(B) ) between BP and AP and performed regression analysis to assess the determinants of P. RESULTS: in both the BP and AP decreased as the number of generic firms increased within these periods. However, this decrease in was significantly less in the AP (median: 0.75 --> 0.68 --> 0.67) than in BP (0.71 --> 0.61 --> 0.53) as the number of generics increased from 1 to 2 to 3, respectively. The regression analysis showed that the price ratio in the AP was higher than that in the BP by 0.05, 0.09, and 0.13 for first, second, and third generic entrant respectively. CONCLUSIONS: Our findings show that the 70/90 regulations not only failed to achieve their goal of lowering the procurement price but instead the opposite occurred. The mandated procurement price became a focal point and resulted in a clustering of prices around the maximum allowable levels with little price dispersion. PMID- 12544551 TI - Licensed nurse staffing and adverse events in hospitals. AB - OBJECTIVE: To examine the changes in licensed nursing staff in Pennsylvania hospitals from 1991 to 1997, and to assess the relationship of licensed nursing staff with patient adverse events in hospitals. DATA SOURCE: A convenience sample of all Pennsylvania, acute-care, hospitals, 1991 to 1997. STUDY DESIGN: The study first describes the percentage change of licensed nursing staff categories in Pennsylvania hospitals from 1991 to 1997. Second, random effects Poisson regressions are used to assess the association of the numbers and proportions of licensed nurses with yearly iatrogenic lung collapse, pressure sores, falls, pneumonia, posttreatment infections, and urinary tract infections. Controls are the yearly number of patients, hospital acuity, and other hospital characteristics. DATA COLLECTION: Secondary data containing patient- and hospital level measures from three sources were recoded to establish the incidence of adverse events, aggregated to the hospital level, and merged to form one data set. PRINCIPAL FUNDING: Licensed nurses' acuity-adjusted patient load increased from 1991 to 1997. Licensed nurse/total nursing staff declined from 1994 to 1997. Greater incidence of nearly all adverse events occurred in hospitals with fewer licensed nurses. Greater incidence of decubitus ulcers and pneumonia occurred in hospitals with a lower proportion of licensed nurses. CONCLUSIONS: This study suggests that licensed nurses' patient load began increasing in the 1990s. Adequate licensed nurse staffing is important in minimizing the incidence of adverse events in hospitals. Ensuring adequate licensed nurse staffing should be an area of major concern to hospital management. Improved measures of nurse staffing and patient outcomes, and further studies are suggested. PMID- 12544552 TI - Patient and community preferences for outcomes in prostate cancer: implications for clinical policy. AB - BACKGROUND: Preferences, or utilities, for health outcomes are central in prostate cancer decision-making. Utilities can be elicited directly from patients using standard techniques, or indirectly, using questionnaires that incorporate preference weights from community members. OBJECTIVES: To evaluate directly elicited and indirectly elicited (questionnaire-derived, community-weighted) utilities for prostate cancer outcomes and the effects of sexual, urinary, and bowel dysfunction on them. MATERIALS AND METHODS: Utilities for the current health of 141 prostate cancer patients, recruited from ambulatory clinics, were elicited directly with the Patient Oriented Prostate Utility Scale, rating scale (PORPUS-U(RS)) and standard gamble (PORPUS-U(SG)) subscales. Patients completed the Health Utilities Index (HUI) and Quality of Well Being Scale (QWB), utility instruments incorporating community preferences, and the UCLA Prostate Cancer Index. RESULTS: Patients' treatments included radical prostatectomy (18%), radiation (60%), and hormonal (42%). Mean utility scores for current health were 0.65 (QWB), 0.79 (PORPUS-U(RS)), 0.80 (HUI), 0.86 (PORPUS-U(SG)). Utility decrements for dysfunction were small (0.08-0.14 [sexual], 0.06 to 0.13 [urinary], and 0.01 to 0.13 [bowel]), and even smaller when adjusted for concomitant changes in other quality of life (QOL) domains. CONCLUSIONS: Patients' directly elicited utilities for their own health were higher than community-derived utilities obtained from HUI and QWB administration to the same patients. HUI scores of these patients were similar to those of age-matched Canadian men. Sexual, urinary, and bowel problems were common but had less impact on overall QOL than reported in previous utility studies. These results weaken the argument that prostate cancer screening and treatment should be limited because of severe and debilitative side effects. PMID- 12544553 TI - Whom do they serve? Community responsiveness among hospitals affiliated with health systems and networks. AB - BACKGROUND: As the US hospital sector becomes more consolidated, concerns have been raised about whether participation in health systems and health networks may reduce community hospitals' response to community health needs. OBJECTIVES: The following were examined: (1) whether freestanding hospitals and system- and network-affiliated hospitals differed in their level of community responsiveness; and (2) how systems and networks affect the level of community responsiveness in community hospitals. METHODS: A cross-sectional design was used. The dependent variables included community orientation, provision of community health services, and Medicaid inpatient load. Independent variables were system/network membership and policy and organizational attributes of the health system/network. RESULTS: With few exceptions, a significantly greater involvement of system and network hospitals was found in providing community health services and inpatient services to Medicaid patients, relative to freestanding hospitals. Community health mission of the system/network and the involvement of the system/network in community partnerships or coalitions were positively related to community orientation in member hospitals. Hospitals affiliated with health systems and hospitals affiliated with more diversified systems or networks tended to provide more community health services. Community health mission of the health system or network was related to greater Medicaid inpatient load in member hospitals. CONCLUSIONS: In general, affiliation with health systems and health networks appears to be positively related to community responsiveness in community hospitals. Research future can examine whether such greater community responsiveness is because of the development and improvement of communication channels among elements of health systems and health networks and the ability of health systems and health networks to build a platform of general, administrative services to link various constituencies. PMID- 12544562 TI - How would you respond to a chemical release? AB - Whether an industrial accident or an act of terrorism, a chemical release can spell disaster. Here you'll learn to respond appropriately, depending on the type and form of chemical involved. PMID- 12544554 TI - Diffusion of new generation antidepressant treatment among elderly diagnosed with depression. AB - RESEARCH OBJECTIVE: This study analyzes the prevalence of new generation antidepressant treatment including selective serotonin reuptake inhibitor (SSRI) use from 1992 to 1997 among the elderly diagnosed with depression, using a large, nationally representative survey of Medicare beneficiaries. Relationships between use of new generation antidepressant treatment and socioeconomic characteristics, physical comorbidity, insurance coverage, and care sector (mental health specialty vs. general health services) are explored. RESEARCH DESIGN: Merged survey data, Medicare claims, and detailed self-reports from the Medicare Current Beneficiary Survey were used to explore use of new generation antidepressant treatment.SUBJECTS Medicare beneficiaries aged 65 and older living in the community, enrolled in fee-for-service Medicare throughout the year and diagnosed with depression. RESULTS: In 1997, among an estimated 1.1 million community dwelling older adults with diagnosis of depression in health care claims, nearly two thirds received antidepressant treatments. Among those diagnosed with depression and treated with antidepressants, the use of new generation antidepressants increased from 35% in 1992 to 77% in 1997. The rates of use increased among all subgroups examined. In the early years after the introduction of these new antidepressant medications (1992-1994), there were socioeconomic disparities in use of these medications, with less use by less educated and poor patients. However, these differences abated over time. CONCLUSIONS: An increasing proportion of elderly treated for depression with antidepressants received the new generation antidepressants. The diffusion of these new medications lagged for those with low socioeconomic status defined by education and income. This diffusion process conforms to a general model of diffusion in which during the initial years following introduction of a new treatment, especially one which is costly, early adopters of the treatment are likely to disproportionately represent those of higher socioeconomic status. PMID- 12544563 TI - A writer, not a nurse. PMID- 12544564 TI - Comforting a confused patient. PMID- 12544565 TI - What's different about this nursing shortage? AB - Driven by unprecedented social and demographic changes, this nursing shortage won't be easily fixed by solutions that worked in the past. Here's what makes this shortage unique- and how you can influence the change shaping your profession. PMID- 12544566 TI - Assessing cardiovascular status. AB - Learn how to evaluate your patient's heart through sight, sound, and touch. PMID- 12544567 TI - How acute renal failure puts the brakes on kidney function. AB - When you understand the various causes and life-threatening effects, you can support your patient's recovery. PMID- 12544569 TI - Inadvertent tracheal decannulation. PMID- 12544572 TI - Infuse management with leadership. PMID- 12544573 TI - Differentiated practice: get beyond the fear factor. PMID- 12544575 TI - Patient safety: new goals set for 2003. AB - Properly apply JCAHO's National Patient Safety Goals within your organization. PMID- 12544576 TI - Empower staff with praiseworthy appraisals. AB - Use a consistent and objective performance appraisal process to empower employees. PMID- 12544577 TI - Take benchmarking to the next level. AB - Reposition and revitalize your organization by teaching staff members to use data for identifying, setting, and directing performance goals. PMID- 12544579 TI - Overcome toxic management. AB - Ineffective, ill-tempered managers hurt employee morale and productivity. Learn what behaviors characterize toxic managers, how they damage an organization, and how to lessen their impact. PMID- 12544580 TI - Uniting to improve end-of-life care. AB - Survey results measure the impact of a national invitational nursing conference on end-of-life care. PMID- 12544581 TI - Achieve ED restraint reduction. AB - Reduce restraint episodes without compromising safety by implementing process oriented, behaviorally based practice tools. PMID- 12544582 TI - IC of CJD in the OR. AB - Review precautions for the safe handling of surgical materials that may be contaminated with infectious agents from patients with Creutzfeldt-Jakob disease. PMID- 12544583 TI - The magnetic pull. AB - Nursing Management continues this popular series that examines the major attributes of organizations earning Magnet status from the American Nurses Credentialing Center. PMID- 12544584 TI - Manage productivity with HPPD. PMID- 12544585 TI - Rapidly identify CHF with POC advances. AB - Testing B-type natriuretic peptide at the point of care can provide a faster, more cost-effective diagnosis of congestive heart failure. PMID- 12544586 TI - NPs stand ready for 2003. PMID- 12544587 TI - New guidelines for heart failure focus on prevention. AB - Heart failure is the only major cardiovascular disease that's increasing in incidence and prevalence in the United States. Because heart failure is a preventable disease, the 2002 heart failure guidelines devise a new staging system that expands the continuum of care to encompass prevention. PMID- 12544590 TI - A look at hypertension in young African-American men. PMID- 12544591 TI - Using cross-cultural definitions of health care. PMID- 12544593 TI - Domestic violence and deployment in US Army soldiers. AB - Although military deployment has been suggested as a possible cause of increases in domestic violence, little is known about it. The purpose of this study was to determine if deployment of 6 months to Bosnia predicted early postdeployment domestic violence. Active duty recently deployed (N = 313) and nondeployed (N = 712) male soldiers volunteered to take an anonymous questionnaire. Deployment was not a significant predictor of postdeployment domestic violence. However, younger soldiers, those with predeployment domestic violence, nonwhite race, and off-post residence also were more likely to report postdeployment domestic violence. The predicted probability of postdeployment domestic violence for a deployed 20-year old, nonwhite soldier with a history of predeployment domestic violence and who lives on-post was.20. For the soldier without a history of predeployment domestic violence, it was.05. Prevention and intervention programs for postdeployment domestic violence shortly after return should target age and persons with a domestic violence history rather than deployment per se. PMID- 12544594 TI - Psychiatric comorbidity and the prevalence of HIV infection in a sample of patients in treatment for substance abuse. AB - This article compares the differences in HIV prevalence and risk behaviors in singly diagnosed patients with substance abuse problems and dually diagnosed patients with comorbid psychiatric and substance abuse problems. The National Treatment Improvement Evaluation Study (NTIES) was used to conduct a cross sectional study on 6593 persons treated for substance abuse disorders. Logistic regression using hierarchal linear modeling (HLM) was utilized to compare risk behaviors of singly and dually diagnosed persons. Compared with singly diagnosed patients, dually diagnosed patients were more likely to share a needle, have sex for money or gifts, have sex with an intravenous drug user, and report being raped. Patients dually diagnosed with psychiatric and substance abuse disorders are an especially high-risk group for HIV infection, in part due to a higher likelihood of engaging in high-risk behavior, and should be targeted for more intense HIV interventions. PMID- 12544595 TI - The links between the Adolescent Dissociative Experiences Scale (A-DES), fantasy proneness, and anxiety symptoms. AB - The purpose of this study was to further examine the psychometric properties of the Adolescent Dissociative Experiences Scale (A-DES). A sample of normal adolescents (N = 331) aged 12 to 18 years completed the A-DES and questionnaires measuring posttraumatic stress disorder (PTSD) symptoms, other anxiety disorder symptoms, and fantasy proneness. Factor analysis indicated that the A-DES, at least in nonreferred youths, is assessing a single dimension of dissociation. Furthermore, A-DES scores are not only significantly related to PTSD symptoms but also to other anxiety disorder symptoms (i.e., generalized anxiety disorder, obsessive-compulsive disorder, and panic disorder) and fantasy proneness. Theoretical and practical implications of these findings are briefly discussed. PMID- 12544596 TI - A clinical interpretation of attitudes and behaviors associated with celebrity worship. AB - The phenomenon of celebrity worship is currently conceptualized as an abnormal type of parasocial relationship, driven by absorption and addictive elements and which potentially has significant clinical sequelae. The authors hypothesize that the three increasingly extreme sets of attitudes and behaviors associated with celebrity worship also partly reflect the three domains of personality discussed in Eysenckian theory. Specifically, celebrity worship for entertainment-social reasons may reflect extraversion personality traits; intense-personal attitudes and behaviors toward celebrities may reflect neuroticism traits; and celebrity worship of a borderline-pathological nature may reflect psychoticism traits. To test this idea, the authors administered the Celebrity Attitude Scale and the Abbreviated Form of the Revised Eysenck Personality Questionnaire to large convenience samples of students (N = 317) and community (N = 290) respondents. Results indicate that celebrity worship is not an uncommon phenomenon. Further, correlational analyses supported predictions and suggest that Eysenckian domains of personality may promote or hinder a person's progression along the continuum of behaviors associated with celebrity worship. PMID- 12544597 TI - Personality disorders and adult attachment dimensions in a mixed psychiatric sample: a multivariate study. AB - The associations between personality disorders and adult attachment dimensions were assessed in a sample of 487 consecutively admitted psychiatric subjects. Canonical correlation analysis showed that two sets of moderately correlated canonical variates explained the correlations between personality disorders and adult attachment patterns. The first and second attachment variates closely resembled the avoidance and anxiety attachment dimensions, respectively. The first personality disorder variate was mainly characterized by avoidant, depressive, paranoid, and schizotypal personality disorders, whereas dependent, histrionic, and borderline personality disorders loaded on the second canonical variate. However, these linear combinations of personality disorders were different from those obtained from principal component analysis. The results extend previous studies linking personality disorders and attachment patterns and suggest the importance of focusing on specific constellations of symptoms associated with dimensions of insecurity. PMID- 12544598 TI - Patterns of neurocognitive deficits and unawareness of illness in schizophrenia. AB - It has been alternately theorized that poor insight in patients with schizophrenia results from deficits in executive function and a preference for denial as a coping strategy. One possibility is there are two distinct groups of persons with poor insight: those with impairments in executive function and those with a generally avoidant coping style. To examine this question, the authors performed a cluster analysis on 64 persons with schizophrenia spectrum disorders on the basis of the PANSS insight and judgment item and executive function assessed with the Wisconsin Card Sorting Test. As predicted, three groups were found: good insight-average executive function (N = 28), poor insight-average executive function (N = 13), and poor insight-poor executive function (N = 23). When self-reported coping styles were compared among groups, as predicted, the poor insight-average executive function group endorsed a significantly greater preference for denial as a coping strategy than the poor insight-poor executive function group, even after controlling for differences in executive function. The coping scores of the good insight group did not differ significantly from either poor insight group. Results suggest that denial may play a role in the unawareness of illness in some persons with schizophrenia who have average executive function. PMID- 12544599 TI - A prospective study on the relationship between self-esteem and functioning during the first year after being hospitalized for psychosis. AB - A recent body of literature on the subjective experience of severe mental illness has reported on the importance of a person's self-esteem. Using a longitudinal design, this study investigates the relationship between self-esteem and functioning during the first year after being hospitalized for psychosis. The findings are based on comprehensive assessments conducted over a 1-year period with 43 persons hospitalized for schizophrenia and related disorders. Results reveal that change in level of self-esteem is significantly related to outcome. These results emphasize the importance of helping patients sustain positive self esteem in the midst of disorder and treatment. PMID- 12544600 TI - Psychiatric morbidity and low self-attentiveness in patients with environmental illness. AB - Controversy surrounds the origin of symptoms attributed to environmental pollutants or widely used chemicals, and the authors believed that a psychiatric evaluation could advance understanding of this contentious condition. They assessed psychiatric morbidity, somatization, and self-attentiveness in patients seen in their Environmental Clinic. Two hundred ninety-five consecutive patients underwent SCID-I and -II interviews and were investigated with self-rating scales for self-attentiveness and somatization. The authors found a high prevalence of mental disorders (66% had a current SCID diagnosis, and 75% had a lifetime SCID diagnosis) and a low level of self-attentiveness, which was not necessarily associated with psychiatric disease. Among patients visiting an Environmental Clinic, mental disorders were common and needed to be diagnosed and treated by standard interventions. Patients who did not meet diagnostic criteria for a psychiatric disorder had relatively low somatization scores and low private self attentiveness. These "externalizers" could benefit from an intervention that teaches them to focus on their internal and emotional lives. In these patients, the authors consider low self-attentiveness a major feature that may act as a pathogenic factor for environmental illness. PMID- 12544601 TI - Retrospective accounts of childhood sexual abuse and current psychological functioning in German and American female undergraduates. PMID- 12544602 TI - Enhanced frontal processing of auditory stimuli in siblings with acute psychosis. PMID- 12544603 TI - Avoiding problems when establishing a student learning experience in perioperative nursing. PMID- 12544604 TI - Clues for clinicians: a case-study approach to educating the renaissance nurse. PMID- 12544605 TI - A process for conducting a curriculum audit. PMID- 12544606 TI - Experiences of psychiatric mental health nursing graduate students in a women's prison. PMID- 12544607 TI - Recruiting students into nursing. PMID- 12544608 TI - Incorporating the experiential learning theory into the instructional design of online courses. AB - Online courses are designed to be asynchronous, interactive, and collaborative. Incorporating the Experiential Learning Theory into the instructional design allows courses to enhance the learning power of learners, empower learners, and challenge learners as they move through a series of planned learning cycles. The author discusses how a learning plan template was developed to facilitate development of assessment, diagnostic, and clinical decision-making skills. PMID- 12544609 TI - An innovative idea: clinical experience with foster care. PMID- 12544611 TI - Elder RNs' perspectives on nursing education: lessons learned. AB - Nursing faculty are challenged with recruiting students and preparing graduates to practice in increasingly complex healthcare settings. The nursing shortage compounds the burden faced by faculty in meeting the needs of the profession. A grounded theory study that explores the perceptions of nurses aged 65 years and older regarding the state of the nursing profession in light of the shortage revealed many interesting aspects of "early" nursing education that are worth revisiting. The author describes the research and uses narrative data to describe what brought elder nurses into nursing, the positive and negative aspects of their education, and the lessons we can learn from their wisdom and experience. PMID- 12544610 TI - Clinical application of the Omaha system with the Nightingale Tracker: a community health nursing student home visit program. AB - The application of computer use in the clinical and educational arena needs to be emphasized for both the improved management of patient data and nursing knowledge. Faculty commitment to automation of home visit documentation records was essential to sustain the trial of implementing the Nightingale Trackers in the clinical area. The Nightingale Tracker is a software program that automates the Omaha system, a community-friendly nursing language that encourages a focus on health promotion. A team approach involving students, faculty, and technical support enabled the automation of the patient record of a home visiting program in a community health nursing course. PMID- 12544612 TI - A university-wide health fair: a health promotion clinical practicum. AB - Can sophomore baccalaureate students in their first clinical experience develop a health promotion activity to meet the needs of a university community? The authors describe how students, focusing on wellness and the nursing process, developed a university-wide health fair; the learning experiences, both planned and serendipitous; the importance of collaboration in health promotion and disease prevention activities; and the promotion of nursing's presence on a university campus. PMID- 12544613 TI - Faculty and student experiences with Web-based discussion groups in a large lecture setting. AB - The exchange of ideas in a discussion format is a more effective way of developing critical thinking in students than a traditional lecture format. Although research has shown that discussion groups are more effective for developing skills in application, analysis, and synthesis of content, it is difficult to implement in a large lecture setting. The author discusses how computer discussion groups were incorporated into a class of 117 nursing students. PMID- 12544614 TI - Using electronic portfolios to measure student achievement and assess curricular integrity. AB - Successful achievement of program outcomes is the primary goal of nursing education programs. Electronic portfolios are a contemporary method by which to measure student achievement, assess curricular efficacy, and evaluate program integrity in nursing education. The authors outline the sequential process of understanding, introducing, and integrating electronic portfolios into a curriculum. PMID- 12544615 TI - Ethical debates: enhancing critical thinking in nursing students. AB - The use of classroom debates can be helpful in teaching ethical content that is often nebulous and difficult for students to comprehend and apply. Debates enhance critical thinking skills through researching issues and developing a stance that can be supported in scientific literature. The authors describe a student debate project involving ethical issues with chronically ill clients. Many students changed their views during the debates. Students evaluated the debates as a positive learning experience. PMID- 12544616 TI - Achieving excellence in end-of-life care. AB - Nurses in practice have verified the need for nursing education curricula to include content related to care of the dying patient. Nurse educators are initiating curricular changes to assure students have the knowledge and attitudes needed to provide quality care at the end of life. The authors discuss one university undergraduate program's development of both a theory and practicum course. PMID- 12544617 TI - Management rights. PMID- 12544618 TI - Nursing references at the point of care. PMID- 12544619 TI - Admission-discharge-teaching nurses: bridging the gap in today's workforce. PMID- 12544620 TI - Essential activities for implementing a clinical information system in public health nursing. PMID- 12544621 TI - Ethical helps and challenges faced by nurse leaders in the healthcare industry. AB - Professional healthcare providers are held responsible for technologic and financial decisions and also for the moral ramifications of their decisions. The authors report the findings of a survey of nurse leaders conducted to determine the key factors that provide help and present challenges as they seek to respond ethically to the dilemmas encountered in the course of their work. Implications for the healthcare industry and the nursing profession are discussed. PMID- 12544622 TI - Nursing documentation time during implementation of an electronic medical record. AB - OBJECTIVE: To determine, within the context of all nursing duties, the amount of time nurses spend on documentation during the implementation of an electronic medical record (EMR) on an intrapartum unit. BACKGROUND: Increased documentation needs during EMR implementation may necessitate increased staffing requirements in an already labor-intensive and demanding environment. METHODS: A work-sampling study was conducted over a 14-day study period, and 18 of 84 (21%) potential 4 hour observation periods were selected. During each period, a single observer made 120 observations and, on locating a specific nurse, immediately recorded that nurse's activity on a standardized and validated instrument. Categories of nursing activities included documentation, bedside care, bedside supportive care, nonbedside care, and nonpatient care. RESULTS: A total of 2160 observations were made. The total percentage of nursing time spent for documentation was 15.8%, 10.6% on paper and 5.2% on the computer. The percentage of time spent on documentation was independently associated with day versus night shifts (19.2% vs 12.4%, respectively). CONCLUSIONS: Despite charting concurrently on both paper and computer, the amount of time spent on documentation was not excessive, and was consistent with previous studies in which neither electronic nor "double charting" occurred. PMID- 12544623 TI - Creating a culturally competent organization: use of the diversity competency model. AB - Recruiting and retaining an adequate nursing workforce is a priority as well as a challenge, and creating an environment that respects every individual's unique differences is key. The authors describe a method for gauging organizational readiness to respond to this challenge. The Diversity Competency Model is used to conduct an in-depth assessment of one nursing organization's diversity initiatives. Leadership commitment, structural linkages, organizational culture, and continuous measurement constitute the Diversity Competency Model assessment. The authors discuss the model and its use. PMID- 12544624 TI - A model for evaluating the context of nursing care delivery. AB - This study used a mixed method to test a model of demands on nurses. Factors were identified that affect demands, and a model was tested using structural equation modeling. Teamwork and expertise had significant effects on nurse-sensitive outcomes. Leadership was a strong determinant of stability and expertise. Workload was not a significant predictor of outcomes, but this is likely due to the difficulty in measuring nursing work. These results provide administrators with actionable recommendations that can be used to reduce the demands on nursing staff. PMID- 12544625 TI - Pharmaceutical gift giving: analysis of an ethical dilemma. AB - When pharmaceutical companies market their products to, and through, healthcare professionals in hospitals and private practice, healthcare professionals face ethical dilemmas in their practice and their organizations. Pharmaceutical companies target nurse practitioners with prescribing privileges. The author describes the ethical dilemma faced by healthcare professionals when friendly salespeople offer tempting gifts. The article outlines cultural responses to gift giving and ethical issues surrounding healthcare professionals' responses to pharmaceutical marketing strategies. Nurse administrators need to acknowledge a growing threat to nursing integrity. Nurse administrators have the power to make and enforce ethical policies that prevent proprietary influences from clouding nursing judgment and contributing to the escalating costs of prescription medications. PMID- 12544626 TI - From past paradigms to future frontiers: unique care delivery models to facilitate nursing work and quality outcomes. AB - As healthcare processes were reengineered in response to managed care, traditional care delivery models were abandoned, resulting in nursing staff dissatisfaction, increased healthcare error, and eroding clinical outcomes. An aging patient population, chronicity of illness, the proliferation of new medical information and technology, severity of illness, and the focus of acute care to "stabilization and transition" necessitate the creation of systems that address changes in nursing work expectations while maximizing available resources. By evaluating unit-specific structure and process criteria and allocation of provider roles, unique, setting-specific care delivery models can be created to facilitate direct and nondirect patient care functions, resulting in improved financial and clinical outcomes. PMID- 12544628 TI - Immunological changes after cancer treatment and participation in an exercise program. AB - PURPOSE: The purpose of this investigation was to evaluate the impact of undertaking peripheral blood stem cell transplantation (PBST) on T-cell number and function, and to determine the role of a mixed type, moderate intensity exercise program in facilitating the recovery of T-cell number and function. METHODS: Immunological measures of white blood cell, lymphocyte, CD3, CD4, and CD8 counts, and CD3 cell function were assessed pretransplant (PI), immediately posttransplant (PII), and 1 month (I1), 2 months (I2) and 3 months (PIII) posttransplant. After PII, 12 patients were divided equally into a control group (CG) or exercise intervention group (EG). RESULTS: Lower total T-cell, helper T cell, and suppressor T-cell counts (P< 0.01), as well as lower T-cell function (P< 0.01), when compared with normative data, were found at PI. More specifically, 88% of the group had CD3, CD4, and CD8 counts that were more than 40%, 20%, and 50% below normal at PI, respectively. Undertaking a PBST caused further adverse changes to the total leukocyte, lymphocyte, CD3, CD4 and CD8 count, and the helper/suppressor ratio. Although CD8 counts had returned to normal by PIII, CD3, CD4, and the CD4 /CD8 ratio remained significantly lower than normative data (P< 0.01), with 66%, 100%, and 100% of the subject group reporting counts and ratios, respectively, below the normal range. CONCLUSION: The PBST patients were immunocompromised before undertaking the transplant, and the transplant procedure imposed further adverse changes to the leukocyte and lymphocyte counts. The leukocyte and CD8 counts returned to normal within 3 months posttransplant; however, the other immunological parameters assessed demonstrated a delayed recovery. Although participation in the exercise program did not facilitate a faster immune cell recovery, neither did the exercise program hinder or delay recovery. PMID- 12544629 TI - Weight lifted in strength training predicts bone change in postmenopausal women. AB - PURPOSE: The aim of this study was to examine the relationship between weight lifted in 1 yr of progressive strength training and change in bone mineral density (BMD) in a group of calcium-replete, postmenopausal women. METHODS: As part of a large clinical trial, 140 calcium-supplemented women, 44-66 yr old, were randomized to a 1-yr progressive strength-training program. Half of the women were using hormone replacement therapy. Three times weekly, subjects completed two sets of six to eight repetitions in eight core exercises at 70-80% of one repetition maximum. BMD was measured at baseline and 1 yr. RESULTS: In multiple linear regression, the increase in femur trochanter (FT) BMD was positively related to total weight lifted (0.001 g.cm (-2)) for a SD of weight lifted, P< 0.01) after adjusting for age, baseline factors, HRT status, weight change, cohort, and fitness center. The weighted squats showed the strongest (0.002 g.cm(-2)) for a SD of weight lifted, P< 0.001), whereas the back extension exhibited the weakest (0.0005 g.cm(-2)) for a SD of weight lifted, P< 0.26) association with change in FT BMD. The amount of weight lifted in the weighted march exercise was significantly related to total body BMD (0.0006 g.cm(-2)) for a SD of weight lifted, P< 0.01). The associations between weight lifted and BMD for the femur neck or lumbar spine were not significant. CONCLUSION: Evidence of a linear relationship between BMD change and total and exercise-specific weight lifted in a 1-yr strength-training program reinforces the positive association between this type of exercise and BMD in postmenopausal women. PMID- 12544630 TI - Bronchoconstriction during cross-country skiing: is there really a refractory period? AB - PURPOSE: The asthmatic airway responds to exercise by bronchodilation (BD) during and bronchoconstriction (BC) after exercise. A refractory period induced by an initial exercise challenge that provides protection against BC during a subsequent exercise bout has also been observed. However, no studies examining during-exercise response or refractoriness during long-duration field exercise by elite athletes have been performed. This study examined airway response and refractoriness during approximately 42-min cross-country ski time trial preceded by a 6- to 9-min 2.5-km high-intensity warm-up ski. METHODS: Eighteen elite athletes cross-country skied seven successive 2.5-km loops. Spirometry was performed pre- and at 5, 10, and 15 min post loop 1; loops 2-7 were treated as a race (XCR) with maneuvers performed within 20 s after loops 2-6 and serially for 15 min after lap 7. RESULTS: Nine of 18 subjects demonstrated a >or=10% fall from baseline in FEV(1) (EIB+): five after lap 1 and four during or after laps 2-7. FEV(1) for EIB+ athletes during XCR was not different from post lap 1 FEV. Only one EIB+ subject demonstrated significant refractoriness. Four EIB+ athletes had a less than 10% fall in FEV after the initial 2.5-km exercise challenge but developed EIB (>or=10% fall) during the subsequent 6 x 2.5 km XCR exercise challenge. FEF(25-75) falls mirrored FEV(1), but demonstrated greater BD during XCR. CONCLUSION: Bronchoconstriction occurs in athletes during prolonged exercise and may thus influence performance. Variability in bronchial hyperresponsiveness onset and the lack of significant refractoriness in our study cohort of athletes is consistent with an exercise bronchoconstrictive dysfunction that is different than frank asthma and is yet to be clearly defined. PMID- 12544631 TI - Differential effects of exercise training on skeletal muscle SERCA gene expression. AB - INTRODUCTION: Exercise training induces significant changes in the performance of skeletal muscle. PURPOSE: To determine whether changes in the gene expression of rat hind-limb muscle sarcoplasmic reticulum Ca2+ -ATPase isoforms (SERCA1a and SERCA2a) in response to either moderate- or high-intensity exercise training underlie the functional remodeling. METHODS: SERCA1a and SERCA2a isoform mRNA expression was determined in predominantly fast-twitch, gastrocnemius muscle and predominantly slow-twitch, soleus muscle with northern analysis. SERCA mRNA was normalized by the 18S rRNA measured in the same sample. RESULTS: Significant increases in the gastrocnemius SERCA2a mRNA expression were observed after both moderate- and high-intensity training. No significant change in SERCA1a expression was found under any conditions. CONCLUSION: These results indicate that both moderate and high intensity exercise increase the relative SERCA2a expression in the gastrocnemius. PMID- 12544632 TI - Nandrolone decanoate enhances hypothalamic biogenic amines in rats. AB - PURPOSE: To identify possible mechanisms for an anabolic-androgenic steroid induced increase in aggressive behavior and work capacity, the levels of some biogenic amines considered to be closely related to a systemic hyper-adrenergic state were measured in selected regions of the brain. METHODS: Wistar male rats were divided randomly into five groups: nontreated (control), oil-vehicle-treated (vehicle) or one of three (therapeutic dose and 10- or 100-fold higher dose) anabolic-androgenic steroid-treated (steroid-1, -2, -3) groups. Rats in the steroid and vehicle groups were given a single dose of nandrolone decanoate or oil vehicle, respectively, one week before tissue sampling. The levels of norepinephrine (NE) and its metabolite, 4-hydroxy-3-methoxyphenylglycol (MHPG), serotonin (5-HT) and its metabolite, 5-hydroxy-indole-3-acetic acid (5-HIAA) were measured in the cerebral cortex, hypothalamus and cerebellum by high-performance liquid chromatography. Immunostaining for c-fos was performed as a confirmation of increased neural activity. RESULTS: The levels of NE and MHPG were increased by approximately 2- and approximately 7-fold in the hypothalamus of the steroid-2 compared with the control and vehicle groups. The levels of 5-HT and 5-HIAA were approximately 40 and approximately 50% higher in the steroid-2 compared with the control and vehicle groups. A significantly higher number of c-fos expressing neurons were observed in the periventricular region of the steroid-2 than the control and vehicle groups, indicating enhanced neuronal activity after nandrolone decanoate treatment. CONCLUSIONS: The present results, combined with previously reported findings of physical performance enhancement after anabolic androgenic steroid treatment, are consistent with the interpretation that elevated levels of adrenergic and serotonergic amines in the hypothalamus could contribute to aggressive behaviors as well as improved physical performance. PMID- 12544633 TI - Muscle architectural characteristics in women aged 20-79 years. AB - PURPOSE: The purpose of this study was to investigate the muscle architectural characteristics, i.e., muscle thickness, pennation angle, and fascicle length, in women aged 20-79 yr. METHODS: A total of 224 sedentary healthy women participated in this study. Muscle thickness and pennation angle of the vastus lateralis (VL), medial gastrocnemius (MG), and triceps brachii (TB) muscles were measured by B mode ultrasonography, and the fascicle length was estimated. RESULTS: Relative muscle thickness of VL and MG decreased significantly with advanced age (VL: P< 0.01, MG: P< 0.05). In addition, the ratio of muscle thickness of VL to that of MG decreased significantly with advanced age (r = -0.206, P< 0.05). The pennation angle decreased significantly with aging for VL (P< 0.001) but not for MG. For TB, no significant age-related changes in relative muscle thickness or pennation angle were observed. There were no significant correlations between age and relative fascicle length (to limb length) in any of the studied muscles. CONCLUSION: These results suggested that the declines in muscle thickness and pennation angle with aging were remarkable in the vastus lateralis muscle and slight in the medial gastrocnemius muscle. For the triceps brachii muscle, the muscle thickness and pennation angle did not decrease with advanced age. Furthermore, no significant age-related changes in relative fascicle length (to limb length) were observed in any of the studied muscles. PMID- 12544634 TI - Interrelationships between muscle structure, muscle strength, and running economy. AB - PURPOSE: The present study was designed to investigate possible differences in running economy (RE) among elite middle-distance runners by examining muscle structure and maximal isometric force (MVC). METHODS: Ten young male runners ran at six different running speeds. During the running bouts, respiratory gases, and blood lactate were measured. Muscle biopsies were obtained from the vastus lateralis muscle for analyzing fiber type distribution, muscle fiber area, myosin heavy chain (MHC) composition, activities of a number of metabolic enzymes (citrate synthase, lactate dehydrogenase, phosphofruktokinase, and 3-hydroxyacyl CoA-dehydrogenase), and titin isoforms. RESULTS: Energy expenditure (EE) increased linearly up to the speed of 6.0 m.s. The relative distribution of the MHC isoforms was MHC I: 67.0%, MHC IIA: 31.5%, and MHC IIX: 1.5%. The present results demonstrated that higher the area of Type II fibers, higher the MVC (r = 0.59, P< 0.05). The amount of MHC II correlated inversely with EE when running close to the competition speed (r = -0.61, P< 0.05). Enzyme activities did not correlate significantly with either RE or EE. Titin analysis revealed that a faster-mobility titin band was observed in all subjects, whereas a lower-mobility titin band was observed only in the most economical runner. CONCLUSION: Differences in RE among homogeneous group of middle-distance runners were observed at various running speeds. This may partly be explained by differences in muscle fiber distribution, MHC composition, and titin isoforms. PMID- 12544635 TI - Does the mechanical work in running change during the VO2 slow component? AB - PURPOSE: The origin of the slow component is not fully understood. The mechanical hypothesis is one of the potential factors, because an increase in external mechanical work with fatigue was previously reported for a constant velocity run. The purpose of this study was to determine whether a change in mechanical work could occur during the development of the VO2 slow component under the effect of fatigue. METHODS: Twelve regional-level competitive runners performed a square wave transition, corresponding to 95% of the speed associated with peak VO2 obtained during an incremental test. The VO2 response was fit with a classical model including two exponential functions. A specific treadmill with three dimensional force transducers was used to measure the ground reaction force. Kinetic work (W(kin)), potential work (W(pot)), external work (W(ext)), and an index of internal work (W(int)) per unit of distance were quantified continuously. RESULTS: During the slow component of VO2, a significant increase in W (P< 0.01), no change in W, and a significant decrease in W and W index (P< 0.05, P< 0.001, respectively) were observed. CONCLUSION: The present study showed that the slow component of VO2 did not result partly from a change in mechanical work under the effect of fatigue. Nevertheless, the decrease in stride frequency (P< 0.001) and contact time (P< 0.001) suggested an alternative mechanical explanation. The slow component during running may be due to the cost of generating force or to alterations in the storage and recoil of elastic energy, and not to the external mechanical work. PMID- 12544636 TI - Use of acetylene breathing to determine cardiac output in young and older adults. AB - PURPOSE: The aims of this investigation were: 1). to establish the day-to-day reproducibility of open-circuit acetylene breathing for measuring exercise cardiac output (Q(c)) in young and older adults; and 2). to compare estimates of Q(c) from open-circuit acetylene breathing with estimates of Q(c) from previously established closed-circuit acetylene rebreathing. METHODS: Twenty men (10 young: 28 +/- 1 yr; 10 older: 61 +/- 1 yr (mean +/- SE)) performed cycle ergometry exercise on 3 separate days. Q(c) was estimated using open-circuit acetylene breathing on 2 d, and closed-circuit acetylene rebreathing on 1 d. RESULTS: Open circuit acetylene breathing was highly reproducible (young: standard error of measurement (SEM) = 1.52 L.min (-1) limits of agreement (LOA) = 0.2 +/- 4.2 L.min (-1), coefficient of variation 6% < CV < 8%, day 2 = (0.9 x day 1) + 2.4, r = 0.90, P< 0.001, r (2)= 0.82; older: SEM = 0.94 L.min (-1), LOA = 0.1 +/- 2.8 L.min (-1), 4% < CV < 10%, day 2 = (1.0 x day 1) + 0, r = 0.91, < 0.001, r(2) = 0.82). Estimates of Q(c) from open-circuit acetylene breathing demonstrated good agreement with closed-circuit acetylene rebreathing (young: SEM = 1.52 L.min ( 1), LOA = 0.9 +/- 4.4 L.min (-1), 5% < CV < 10%, open-circuit = (1.0 x closed circuit) + 1.5, r = 0.89, < 0.001, r (2) = 0.79; older: SEM = 1.13 L.min (-1), LOA = 0.1 +/- 3.2 L.min (-1), 5% < CV < 9%, open-circuit = (0.9 x closed-circuit) + 1.6, r = 0.88, < 0.001, r(2) = 0.78). CONCLUSION: These results demonstrate that open-circuit acetylene breathing provides reproducible measurements of Q(c) during exercise that demonstrate good agreement with values obtained from the acetylene rebreathing procedure in young and older healthy men. PMID- 12544637 TI - Resistance exercise, the Valsalva maneuver, and cerebrovascular transmural pressure. AB - PURPOSE: To examine the acute effects of resistance exercise (RE) performed without a Valsalva maneuver (VM) versus a VM performed alone on systolic pressure, intracranial pressure (ICP), and cerebrovascular transmural pressure (CVTMP) (i.e., the important pressure that stresses the cerebral arterial and aneurysmal walls and calculated as systolic pressure minus ICP). METHODS: The subjects for this study consisted of seven (mean +/- SD, Age: 39 +/- 14 yr) fully alert, cooperative, and clinically stable individuals who recently underwent various neurosurgical operations. Heart rate, systolic pressure, ICP, and CVTMP were obtained at rest, during a VM, and during submaximal (8.0 +/- 3.0 kg performed for 18 +/- 10 repetitions) unilateral bicep curl RE. RESULTS: The VM resulted in a significantly greater increase in ICP (VM: 31 +/- 14 mm Hg vs RE: 16 +/- 7 mm Hg, P< 0.05) with a concomitant decrease in CVTMP (VM: 106 +/- 22 mm Hg vs RE: 132 +/- 14 mm Hg, P< 0.05) compared with unilateral bicep curl RE. CONCLUSIONS: Unilateral bicep curl RE results in a greater increase in CVTMP compared to a VM performed alone. PMID- 12544638 TI - Effect of creatine ingestion on glucose tolerance and insulin sensitivity in men. AB - PURPOSE: This study investigated whether acute (5 d) and/or short-term (28 d) creatine (Cr) ingestion altered glucose tolerance or insulin action in healthy, untrained men (aged 26.9 +/- 5.7 yr; SD). METHODS: Subjects were randomly allocated to either a Cr ( N= 8) or placebo group (N = 9) and were tested in the control condition (presupplementation), and after 5 and a further 28 d of supplementation. The Cr group ingested 20 g and 3 g.d (-1) of Cr for the first 5 and following 28 d, respectively. The placebo group ingested similar amounts of glucose over the same time period. During each testing period, subjects underwent an oral glucose tolerance test (OGTT) to determine insulin sensitivity, and six subjects from each group underwent a muscle biopsy before each OGTT. RESULTS: Cr supplementation resulted in an increased (P< 0.05) muscle TCr content after both the acute and short-term loading phase compared with placebo. Neither acute nor short-term Cr supplementation influenced skeletal muscle glycogen content, glucose tolerance, or measures of insulin sensitivity. CONCLUSIONS: These findings demonstrated that acute Cr supplementation (20 g.d(-1) for 5 d) followed by short-term Cr supplementation (3 g.d(-1) for 28 d) did not alter insulin action in healthy, active untrained men. PMID- 12544639 TI - Human albumin synthesis is increased by an ultra-endurance trial. AB - PURPOSE: The purpose of this study was to determine whether an ultra-endurance event is a strong stimulus to increase albumin synthesis involved in the process of intravascular albumin mass increase associated with transient hypervolemia. METHODS: The albumin synthetic rate was measured in six young men, 3 d before (C) and on the 1st (R1) and 8th (R8) days of the recovery from an ultra-endurance trial (5 h daily for 4 d). Albumin fractional (FSR) and absolute (ASR) synthetic rate were determined using a primed-constant infusion of [1(-13) C] leucine. Plasma volume (PV) using Evans Blue dye dilution and total body water (TBW) using bioelectrical impedance analysis were measured on C, R1, and R8. RESULTS: On R1 as compared with C: 1). PV (+23.3 +/- 3.2%; P/= 80 cm), and 58 had WCs less than 80 cm. The two groups were comparable in demographic variables and body mass index (BMI). The women with larger WCs were more insulin-resistant than their counterparts. The metabolic risk scores were significantly higher in women with abdominal obesity than in those without it. The results from the GLM showed that WC was an independent predictor of insulin resistance and metabolic risk scores after controlling for demographic variables (0.06- and 0.29-unit increases in homeostasis model assessment for insulin resistance and metabolic risk scores per 1 cm change in WC). Moreover, BMI neither correlated with metabolic risk scores nor affected the WC effects on insulin resistance and metabolic risk scores in the GLM. CONCLUSIONS: Postmenopausal Chinese women with abdominal obesity may carry higher metabolic cardiovascular risk than those without it. It is WC, not BMI, that predicts metabolic cardiovascular risk factors in these women. PMID- 12544681 TI - Hot flushes in a male population aged 55, 65, and 75 years, living in the community of Linkoping, Sweden. AB - OBJECTIVE: Hot flushes are as common in castrated men as in menopausal women. We investigated whether hot flushes exist in a normal aging male population and to what extent. DESIGN: A questionnaire was sent to all men living in Linkoping, Sweden, who were 55, 65, and 75 years old ( = 1,885). The questionnaire asked for demographic data, medical history, mood status, medication, castrational therapy, and smoking, exercise, and alcohol habits, among other items. We asked specifically for current hot flushes unrelated to exercise or a warm environment. RESULTS: Of the questionnaires received, 1,381 were eligible for evaluation; 33 were analyzed separately because these men had been castrated. Hot flushes of any frequency were reported by 33.1% of noncastrated men, 4.3% reported flushes at least a few times per week, and 1.3% reported daily flushes. Half of the men reporting flushes were also bothered by them, ie, almost every sixth man in total. We found a relation between occurrence of hot flushes and other symptoms thought to be related to low testosterone concentration, such as decreased muscle strength or endurance, decreased enjoyment of life, sadness or grumpiness, and lack of energy ( < 0.05). CONCLUSIONS: Hot flushes occur in one third of a population of noncastrated older men, approximately half of whom consider flushes as bothersome. Neither the mechanisms nor whether the symptoms would respond to testosterone supplementation is known. Androgen substitution to treat symptoms possibly related to a male climacteric is still controversial. Studies are needed to evaluate the needs for and the effects of androgen treatment on vasomotor symptoms. PMID- 12544682 TI - Intranasal hormone replacement therapy. AB - Although the optimal route of delivery for hormone replacement therapy has not yet been determined, desirable qualities would include good efficacy, easy administration, minimal side effects, and optimal therapeutic profile. This would potentially serve to improve patient compliance and satisfaction. The intranasal route has been evaluated for the administration of menopausal hormones and seems to fulfill these requirements. The intranasal route would also seem to be a viable alternative for drugs that are poorly absorbed after ingestion by avoiding hepatic first-pass elimination. The intranasal route is, therefore, innovative for the delivery of natural sex steroids in postmenopausal women receiving hormone replacement therapy. Early studies demonstrate that it is safe, effective, and acceptable to postmenopausal women. In addition, the nasal administration of a combination of estradiol and progesterone would seem to be an attractive way to deliver hormones to nonhysterectomized postmenopausal women. Providing alternative routes of administration may also enhance compliance. PMID- 12544683 TI - Estrogen-related pancreatitis in the setting of normal plasma lipids: case report. AB - A case of recurrent acute pancreatitis occurring in conjunction with intermittently used estrogen therapy over 7 years is presented. Estrogens have been associated with pancreatitis in the past, and a causal link involving elevated plasma triglyceride levels has usually been assumed. The current case is distinct in that the plasma lipids of this patient have always been normal, suggesting that a different mechanism must be responsible for the onset of pancreatitis. PMID- 12544684 TI - Tibolone was associated with an increase in 7 alpha-methyl-ethinylestradiol. PMID- 12544685 TI - Tibolone was associated with an increase in 7 alpha-methyl-ethinylestradiol. PMID- 12544686 TI - Segmental colitis: so what? AB - The existence of chronic mucosal inflammation, confined to sigmoid colons that bear diverticula, is termed 'segmental colitis'. This condition often mimicks inflammatory bowel disease at histological examination. The observed rectal sparing suggests a possible form of Crohn's disease, but no other similarities between segmental colitis and Crohn's colitis are detectable. Medical treatment for segmental colitis, empirically carried out with drugs such as sulfasalazine and mesalazine, is mostly successful and, when surgery is required, post operative recurrences are infrequent. Although the existence of segmental colitis as a true clinical entity remains questionable, it appears unlikely that this condition represents an atypical form of inflammatory bowel disease. PMID- 12544687 TI - Living donor liver transplantation in adults. AB - Adult-to-adult living donor liver transplantation (LDLT) using right hemi-liver has become a promising treatment modality for patients with end-stage liver disease. The rapid adoption of the procedure has been triggered mainly by the significant gap between available cadaver grafts and the number of patients on the waiting list for liver transplant. Since the arguments against LDLT focus only on the safety of the donor, the demonstration that the donor operation can be performed with minimal morbidity will make it more ethically acceptable. The advantages of LDLT are the possibility of performing an elective operation, access to a graft in best condition, and the possibility of lowering the likelihood of death while waiting for a suitable organ. As well as the standard indications for liver transplantation, LDLT opens up the possibility of treatment of patients with borderline indications. Further improvement of surgical and medical technology, careful long-term follow-up of donors and recipients, and profound analysis of socioeconomic aspects are essential issues for the transplantation community. PMID- 12544688 TI - Adult-to-adult living donor liver transplant: UK experience. AB - BACKGROUND: Adult-to-adult living donor liver transplantation (ALDLT) is being adopted widely in the USA and mainland Europe, fueled by the increasing waiting lists for cadaver organs. The present report describes the first UK experience with the procedure in patients from overseas who have the lowest priority for cadaver organ allocation. METHODS: The 16 patients seen over the period November 1998 to March 2002 had end-stage cirrhosis from chronic hepatitis C virus (HCV) or hepatitis B virus (HBV) infection (13 cases), with single instances of cryptogenic cirrhosis, secondary biliary cirrhosis and alcoholic liver disease. Grafts were left lobe in the first two recipients and right lobe in the subsequent 14 recipients, donated by nine sons/daughters and seven brothers/sisters. RESULTS: Twelve of the 16 recipients did well. The four recipients who died had recurrent sepsis; two of these died following hepatic arterial occlusion, and in three major surgical factors were present before transplantation. Serial computed tomography (CT) measurements in the survivors showed regeneration of the grafted lobe with final volumes reaching in each case the calculated standard liver volume for body size. In the donors, liver function tests had returned to normal by day 7-14, with rapid regeneration of the remaining lobe, although the final size attained that estimated before donation in only four donors. CONCLUSIONS: ALDLT, although requiring considerable facilities and organization, can give good results for both recipient and donor. As with cadaver grafts, outcome in the recipient if the larger right lobe is used is dependent on surgical risk factors and the severity of clinical decompensation before transplantation. Measures to ensure the safety of the donors remain the main concern. PMID- 12544689 TI - Efficacy and safety of a prothrombin complex concentrate with two virus inactivation steps in patients with severe liver damage. AB - OBJECTIVE: To evaluate the efficacy and safety of intravenous infusions of an improved prothrombin complex concentrate (PCC) formulation. PATIENTS AND METHODS: Twenty-two adults with haemostatic defects due to severe liver disease (Quick's test 50%), which required rapid haemostasis because of bleeding or before urgent surgery or invasive intervention. Laboratory follow-up, including the response and in-vivo recovery of the substituted coagulation factors II, VII, IX and X and protein C took place before, then 10 min, 30 min and 60 min after PCC substitution. Clinical efficacy (avoidance or cessation of bleeding) was assessed using a scale ranging from 'very good' to 'none'. RESULTS: Patients received a median PCC dose of 25.7 IU/kg. The response of factor IX and protein C was 1.2 1.4 (IU/dl)/(IU/kg), the in-vivo recovery was 49.7-57.4%, and the Quick's test increased from 39% to a maximum of 65%. Levels of activation markers of the coagulation system factor VIIa, prothrombin fragment 1 + 2 and thrombin antithrombin complex (TAT) increased, but without evidence of any thromboembolic events. Clinical efficacy was judged as 'very good' in 76% of patients after the first (n = 21) treatment. There were no changes in serological status regarding transmission of HIV, hepatitis A virus, hepatitis B virus and hepatitis C virus. No PCC-related adverse reactions occurred. CONCLUSIONS: The infusion of pasteurized, nanometre-filtered PCC is an effective, well-tolerated method of correcting prothrombin complex deficiency in patients with severe liver disease with haemorrhage, or before an urgent surgical or invasive diagnostic intervention. PMID- 12544690 TI - The effect of transcutaneous nerve stimulation on intraductal biliary pressure in post-cholecystectomy patients with T-drainage. AB - OBJECTIVE: To evaluate the effects of transcutaneous electrical nerve stimulation (TENS) on intraductal biliary pressure (IDP) in basal conditions and after intravenous morphine and oral meal stimulation. DESIGN AND METHODS: Fifteen patients (5 male, 10 female) aged 31-83 years (mean 61.5 +/- 13.7 years) with prior cholecystectomy and residual in situ T-tube were examined. Final radiographs excluded any organic abnormalities. The study consisted of three sessions. On the first day (session 1), after the initial manometric intraductal pressure was measured for 15 min, TENS (using a PRO-TENS pocket stimulator) was applied for 15 min. Measurement was continued for 15 min after termination of TENS. The measurement was performed using a water-perfused manometry system (Synectics Medical, Stockholm, Sweden) by a triple-channel manometric catheter inserted into the common bile duct through a T-drain. On the following day (session 2), the protocol was similar except that, after basal IDP measurement, morphine hydrochloride 0.08 mg/kg was injected intravenously 10 min before TENS. On the third day (session 3), after basal measurements were taken, patients were given a standard test meal and the IDP was recorded continuously for 45 min. To estimate the effects of the stimuli applied, absolute intraductal pressure changes were analysed. RESULTS: In session 1, TENS reduced basal IDP in all patients by a mean of 3.95 +/- 1.6 mmHg. In 13 patients, 15 min after cessation of TENS a further decrease in IDP was observed. In two patients, termination of TENS was followed by a rebound increase in IDP; however, it did not reach the initial value (mean total decrease 5.05 +/- 2.25 mmHg). In session 2, administration of morphine produced an evident increase in IDP in all subjects by 6.9 +/- 2.7 mmHg. TENS decreased IDP in 13 patients. In two patients, TENS initially failed to lower elevated pressure, but it appeared several minutes after the end of stimulation. In 13 patients, the final IDP values were lower than the baseline pressures. In session 3, after administration of a test meal, IDP decreased within 30-40 min by a mean of 4.89 +/- 1.29 mmHg. CONCLUSIONS: TENS decreased basal as well as elevated IDP in the majority of the T-drain patients studied. The effect of TENS persisted after its termination. Elevated IDP is believed to be responsible for pain in patients with sphincter of Oddi dysfunction (SOD). Therefore, we think that TENS can be used effectively and safely as an optional therapeutic method in the treatment of biliary dyskinesia. PMID- 12544691 TI - Impact of clarithromycin resistance and CYP2C19 genetic polymorphism on treatment efficacy of Helicobacter pylori infection with lansoprazole- or rabeprazole-based triple therapy in Japan. AB - OBJECTIVE: Helicobacter pylori treatment failure is thought to be due mainly to polymorphic cytochrome P450 2C19 (CPY2C19) genetic polymorphism, associated with proton pump inhibitor metabolism, and antimicrobial susceptibility. This report has ascertained which was more important, CPY2C19 polymorphism or antimicrobial susceptibility, when using 1-week lansoprazole-based or rabeprazole-based triple therapy in Japan. DESIGN: An open, randomized, parallel group study. SETTING: One hundred and forty-five subjects with H. pylori-positive gastritis or peptic ulcers were randomly assigned to receive 30 mg lansoprazole twice daily (LAC group), 10 mg rabeprazole twice daily (RAC20 group), or 20 mg rabeprazole twice daily (RAC40 group), with 1000 mg amoxicillin twice daily and 400 mg clarithromycin twice daily for 1 week. Antimicrobial resistance testing was performed by E-test. More than 4 weeks after completion of treatment, H. pylori status was assessed by 13C-urea breath test, histology, and culture. RESULTS: Cure rates expressed as intention-to-treat and per-protocol analyses, respectively, were 79.6 and 83.0% with LAC, 85.4 and 89.1% with RAC20, and 83.3 and 88.9% with RAC40. In the case of clarithromycin-sensitive strains, the cure rates were more than 97%, regardless of CPY2C19 polymorphism. However, treatment succeeded in only one out of 16 clarithromycin-resistant strains. CONCLUSIONS: The key to successful eradication of H. pylori, using lansoprazole or rabeprazole with clarithromycin and amoxicillin, is clarithromycin susceptibility, not CPY2C19 polymorphism. PMID- 12544692 TI - Mucosal changes in the gastric remnant: long-term effects of bile reflux diversion and Helicobacter pylori infection. AB - OBJECTIVE: Bile reflux is thought to be responsible for reflux gastritis and stump carcinoma occurring after partial gastrectomy for peptic ulcer. Gastritis and gastric carcinoma are also correlated with Helicobacter pylori. The aim of this study was to investigate whether diversion of enteric reflux and the presence of H. pylori infection alter long-term histological developments in the gastric remnant. METHODS: Twenty-nine patients partially gastrectomized for peptic ulcer were reoperated on with re-resection and a Roux-en-Y reconstruction because of reflux gastritis (12 patients) or severe dysplasia/early gastric cancer (17 patients). The resected specimens and subsequent biopsies from the new anastomotic region taken at endoscopies 5-17 years after reoperation were evaluated regarding the presence of H. pylori, the grade of active and non-active chronic gastritis, and the premalignant changes--atrophy, intestinal metaplasia and dysplasia. RESULTS: A progression of active chronic gastritis, atrophy, intestinal metaplasia and dysplasia was seen after re-resection and Roux-en-Y reconstruction. Non-active chronic gastritis remained unchanged. The development was, in general, independent of H. pylori infection. CONCLUSIONS: Enteric reflux may perhaps induce a histological transformation of the gastric mucosa that cannot be reversed, even if the reflux is diverted. In our study, H. pylori infection had no impact on the histological development. Factors other than enteric reflux and H. pylori infection might also be of importance. PMID- 12544693 TI - Alteration of glutathione S-transferase levels in Barrett's metaplasia compared to normal oesophageal epithelium. AB - OBJECTIVE: Oesophageal cancer associated with the premalignant condition Barrett's oesophagus has increased in incidence over the last few years. Phase II detoxifying enzymes, including glutathione S-transferases (GSTs) protect the mucosa from carcinogens, which can cause oxidative damage to cells. Therefore, a reduction in these anti-oxidant enzymes can increase the risk of carcinogenesis. The aim of this study was to compare the extent of GST expression in normal oesophageal tissue, Barrett's oesophagus and oesophageal adenocarcinoma. DESIGN: Antibodies raised against GST alpha, GST mu, GST pi and microsomal GST were used to identify expression of these proteins in tissue sections. METHOD: Paraffin embedded sections were stained using standard immunohistochemical techniques to demonstrate the pattern of expression of GST proteins in biopsy specimens. Twelve sections of Barrett's metaplasia and an equal number of specimens from normal oesophageal tissue were examined, together with sections from adenocarcinoma and normal gastric mucosa. RESULTS: Expression of the GST enzymes appeared to be reduced in Barrett's tissue compared to normal oesophageal tissue. Nuclear staining featured in some of the normal tissue sections, but not in Barrett's tissue. CONCLUSION: The reduction in GST expression suggested in Barrett's tissue is an interesting finding, as it is possible that reduced expression of these detoxifying enzymes may contribute to the risk of development of adenocarcinoma in Barrett's mucosa. PMID- 12544694 TI - Inflammation at the cardio-oesophageal junction: relationship to acid and bile exposure. AB - BACKGROUND: The aetiology of inflammation in cardiac mucosa at the gastro oesophageal junction (carditis) is unclear, although gastro-oesophageal reflux has been suggested. OBJECTIVES: To correlate histological features of carditis with oesophageal acid exposure (gastro-oesophageal reflux) and proximal gastric bile exposure (duodenogastric reflux) in patients with symptoms of gastro oesophageal reflux disease (GORD). METHODS: Sixty-six patients with reflux symptoms underwent endoscopy with biopsy, oesophageal manometry, 24-h oesophageal pH testing and 24-h proximal gastric Bilitec 2000 testing. Inflammation in glandular mucosa was assessed using the updated Sydney System. Fifteen healthy volunteers underwent pH and Bilitec 2000 testing and served as controls. RESULTS: There was no correlation between either the presence or histological grade of carditis and oesophageal acid exposure or proximal gastric bilirubin exposure. Patients with reflux symptoms had as much duodenogastric reflux into the proximal stomach as did control subjects. CONCLUSIONS: We were unable to establish either gastro-oesophageal or duodenogastric reflux as the predominant cause of inflammation in cardiac mucosa. PMID- 12544695 TI - Abnormal levels of neuropeptide Y and peptide YY in the colon in irritable bowel syndrome. AB - OBJECTIVE: To assess the levels of gut peptides involved in gastrointestinal motor, secretory and sensory function in colonic biopsies in irritable bowel syndrome (IBS) patients and healthy controls. METHODS: We studied 34 patients with IBS and 15 subjects without gastrointestinal symptoms. The predominant bowel pattern in the IBS patients was constipation in 17 patients (IBS-C) and diarrhoea in 17 patients (IBS-D). With radioimmunoassay, the levels of vasoactive intestinal peptide (VIP), substance P, neuropeptide Y (NPY) and peptide YY (PYY) were analysed in biopsies from the descending colon and ascending colon obtained during colonoscopy. RESULTS: The IBS patients had lower levels of PYY in the descending colon than the controls, but the levels in the ascending colon did not differ. The NPY levels were lower in IBS-D than in IBS-C, both in the ascending colon and in the descending colon. Low levels of VIP were more common in IBS patients, but mean levels did not differ between groups. No group differences were observed for substance P. The levels of VIP, substance P and NPY were higher in the ascending colon than in the descending colon, whereas the opposite pattern was seen for PYY. CONCLUSION: IBS patients demonstrate lower levels of PYY in the descending colon than controls. Colonic NPY levels differ between IBS subgroups based on the predominant bowel pattern. These findings may reflect the pathophysiology of IBS and the symptom variation within the IBS population. PMID- 12544696 TI - 6-Thioguanine seems promising in azathioprine- or 6-mercaptopurine-intolerant inflammatory bowel disease patients: a short-term safety assessment. AB - OBJECTIVE: 6-Mercaptopurine (6-MP) and azathioprine (AZA) have proven efficacy in the treatment of inflammatory bowel disease (IBD). However, adverse events leading to discontinuation may occur in 10-20% of patients. The efficacy of AZA and 6-MP is based on formation of their active metabolites, the 6 thioguaninenucleotides (6-TGNs). Therefore, 6-thioguanine (6-TG), an agent leading more directly to the formation of 6-TGNs and until recently used only in patients suffering from leukaemia, may be an alternative in AZA or 6-MP intolerance. The purpose of our study was to assess the short-term safety of 6 TG. METHODS: Thirty-two IBD patients with previously established AZA or 6-MP intolerance were treated with 6-TG in doses of 20 mg (n = 19) or 40 mg (n = 13) once daily. Safety parameters were obtained at 0, 1, 2, 4 and 8 weeks after start of medication. Primary outcome measures were the ability to tolerate 6-TG and the occurrence of adverse events. Secondary outcome definitions included laboratory parameters. RESULTS: Twenty-six (81%) patients were able to tolerate 6-TG during the first 8 weeks. In three of six patients, side effects leading to discontinuation were probably (n = 2) or obviously (n = 1) related to 6-TG. No clinically relevant haematological events or hepatotoxicity occurred in the observed period. Steady-state 6-TG levels were significantly higher with 40 mg once daily (1621 +/- 828 picomol/8 x 10(8) red blood cells (RBC)) than with 20 mg once daily (937 +/- 325 picomol/8 x 10(8) RBC; n = 0.001). CONCLUSIONS: 6-TG treatment seems promising in AZA- or 6-MP-intolerant IBD patients. However, long term safety and efficacy have yet to be determined. PMID- 12544697 TI - Assessment of orocaecal transit time in different localization of Crohn's disease and its possible influence on clinical response to therapy. AB - BACKGROUND: A study on orocaecal transit time (OCTT) in patients with different localizations of Crohn's disease (CD) is not available. Because slow-release drug formulations are increasingly available for the treatment, there is a concrete risk that delayed OCTT may impair the efficacy of these formulations. AIMS: We investigated OCTT before and after therapy using lactulose H2-breath test and we studied whether OCTT can influence the clinical response to therapy with slow release mesalazine formulations in adult CD patients.(2) PATIENTS AND METHODS: We studied 45 adult patients with non-obstructive CD and Crohn's Disease Activity Index (CDAI) <200 (29 men, 16 women; mean age 42 years, range 22-73 years). Twenty patients had ileocolonic, 16 colonic and 9 ileal localization of CD. The control group consisted of 20 healthy subjects (13 men, seven women; mean age 53 years, range 22-71 years). After OCTT assessment, 29 patients were treated with time-dependent mesalazine 3.6 g/day, while 16 patients were treated with pH dependent mesalazine 3.6 g/day. If bacterial overgrowth was detected, the patients were also treated with rifaximin 800 mg/day for 7 days. RESULTS: OCTT was delayed (120 min, range 115-210 min) in 30 of the 45 CD patients (67%). Four patients (9%) showed bacterial overgrowth, while OCTT was regular (82.5 min, range 75-90 min) in 11 patients (24%). In the control group, the mean OCTT was 88.2 min (range 75-135 min); (P<0.01). OCTT was more prolonged in ileal localization (182.2 min, range 150-210 min), rather than in patients with ileocolonic (122 min, range 75-180 min) or colonic (106 min, range 75-150 min) localization of CD; (P<0.01). Thirty-nine patients showed normal OCTT after starting therapy (83 min, range 75-105 min), while OCTT remained slightly delayed in the remaining patients (110 min, range 105-115 min); (P<0.01). CDAI was <100 (mean value 83) in all patients with reduction of OCTT to normal value 4 months after starting therapy, while it was >140 (mean value 143) in patients with a slight reduction but not normalization of OCTT respectively (five patients with ileal and one with ileocolonic localization of CD) with a statistically significant correlation between OCTT and CDAI (P<0.01). CONCLUSIONS: This study shows clearly for the first time that OCTT is not only delayed in patients with active CD, but also that it is prolonged in ileal and ileocolonic rather than colonic localization of CD. Moreover we obtained these results using a simple, sensitive, non-invasive and repeatable method, namely, a lactulose hydrogen breath test. PMID- 12544699 TI - Gross chylous ascites in cirrhosis with massive portal vein thrombosis: diagnostic value of lymphoscintigraphy. A case report and review of the literature. AB - Chylous ascites is an uncommon condition, which could be due to various causes. We report a case of gross chylous ascites in a patient with cirrhosis and portal vein thrombosis. It is confirmed that gross chylous ascites in a patient with cirrhosis and portal vein thrombosis heralds an ominous prognosis for the patient. Results also demonstrate that common therapeutic interventions confer minimal benefit to the patient, whose survival may be limited to a few months. The use of lymphoscintigraphy as a convenient method for diagnostic exploration of the chylous ascites is emphasized, as it does not lead to complications or adverse effects, and can be readily repeated as needed. PMID- 12544698 TI - Activity and expression of human telomerase in normal and malignant cells in gastric and colon cancer patients. AB - BACKGROUND: The reactivation of telomerase is believed to play an important role in immortalization and carcinogenesis. OBJECTIVE: To investigate the expression of three components of the telomerase complex (hTR, hTERT and TP1), along with telomerase activity in malignant and normal cells. METHODS: Cells were isolated from gastric and colon cancer, and from normal mucosa from the stomach and colon of participating patients. Expression of hTERT, hTR and TP1 has been studied by the reverse transcriptase polymerase chain reaction (PCR) technique. The telomerase repeat amplification protocol and PCR enzyme-linked immunosorbent assay were used for analysis of telomerase activity. RESULTS: All telomerase components were consistently expressed in colon and gastric cancer cells. Neoplastic RNA produced consistently very strong amplification signals either for hTR, hTERT or TP1. The expression of hTR was observed in RNA isolated from all normal mucosa samples and from peripheral blood lymphocytes. The expression of TP1 and hTERT has been found in the majority of normal cells; however, the amplification signals produced were usually much weaker than in malignant cells. The limiting dilution experiments indicated that the cancer cells have at least 100-fold higher telomerase activity and at least 25-fold higher TP1 and hTERT expression in comparison to normal cells. CONCLUSIONS: It can be concluded that all the cancer cells tested have higher telomerase expression and activity than normal cells. Therefore, telomerase can be a good cancer marker, provided that quantitative analysis is carried out. PMID- 12544700 TI - Liver failure in adult Still's disease during corticosteroid treatment. AB - Adult Still's disease is a well-characterized rheumatic disorder of unknown origin, which may affect multiple organs and may have a fatal course. However, liver failure has rarely been described in adult Still's disease. We present the case of a 25-year-old woman who was admitted with acute liver failure 2 years after the start of symptoms (arthritis, fever, sore throat) of a yet undefined rheumatic disease. She had been treated with prednisolone for 2 months before admission. The diagnosis of adult Still's disease was made in accordance with well-established criteria. Other causes of liver failure were excluded. Withdrawal of prednisolone did not affect the course of liver disease. Ursodeoxycholic acid therapy was started when the patient slowly began to recover. To the best of our knowledge, this is the first case of adult Still's disease reported in which hepatic failure developed when other symptoms were well controlled by corticosteroid treatment. PMID- 12544701 TI - Kayser-Fleischer like rings in alcoholic liver disease: a case report. AB - Kayser-Fleischer rings are brown pigmented rings that run along the periphery of the cornea. Situated in Descemet's membrane and being comprised of granules of deposited copper they have traditionally been thought of as pathognomic of Wilson's disease. However, they can also be seen in other forms of liver disease. We document a case of Kayser-Fleischer like rings occurring in alcoholic liver disease--a previously unreported association. PMID- 12544702 TI - Acute clinical hepatitis by immune restoration in a human immunodeficiency virus/hepatitis B virus co-infected patient receiving antiretroviral therapy. AB - We describe a human immunodeficiency virus (HIV)-infected patient with chronic active hepatitis B. The patient presented with acute clinical hepatitis during highly active antiretroviral therapy concomitant with immunovirological improvement of HIV infection. The hepatitis resolved within a few weeks despite the pursuit of antiretroviral drugs. Hepatitis B tests, which remained unchanged after two cycles of interferon alfa therapy and continuous administration of lamivudine, were improved with the clearance of hepatitis B virus (HBV) early antigen (HBeAg), the detection of antibody to HBV early antigen (HBeAb) and the undetectability of serum HBV DNA. Several months after this episode, HBV and HIV tests are still controlled. PMID- 12544703 TI - Recurrence of insulin resistant metabolic syndrome following liver transplantation. AB - Insulin resistant metabolic syndrome is a major clinical disorder including hyperlipidaemia, hypertension, impaired glucose tolerance and/or type 2 diabetes and central obesity, which are well established cardiovascular risk factors. We report the case of a 61-year-old woman who developed severe hypercholesterolaemia and hypertriglyceridaemia after liver transplantation. In her forties she had hypertension, mixed hyperlipidaemia, mild hyperglycaemia and moderate abdominal obesity, suggesting the presence of the metabolic syndrome. She had liver enzyme elevation and severe steatosis and hepatomegaly at ultrasonography. At age 52, cryptogenic liver cirrhosis was diagnosed and rapidly progressing liver failure developed. In 1992 she underwent liver transplantation. Seven years after transplant the patient had abdominal obesity, high blood pressure, marked hypercholesterolaemia, hypertriglyceridaemia and moderate elevation of alanine aminotransferase. She also had impaired glucose tolerance and markedly increased basal and post-glucose load plasma insulin levels. Steatohepatitis was demonstrated by serial liver biopsies. This is the first case that reports the recurrence of the metabolic syndrome following liver transplantation. We postulate that metabolic syndrome may have promoted fatty liver and subsequent progression to end stage liver disease. We also stress the need for careful management of the metabolic syndrome in order to decrease the long-term risk for cardiovascular disease. PMID- 12544704 TI - Macro-AST as cause of unknown hypertransaminasaemia. PMID- 12544705 TI - Paracetamol poisoning and the international normalized ratio. PMID- 12544706 TI - Bilateral vertebral artery occlusion resulting from giant cell arteritis: report of 3 cases and review of the literature. AB - Giant cell arteritis (GCA) is known to affect the extracranial part of the vertebral arteries. Bilateral vertebral artery occlusion (BVAO) is a rare but serious neurologic condition. We report 3 patients with autopsy-proven (2 patients) or clinically diagnosed (1 patient) GCA causing BVAO. A review of the literature concerning BVAO revealed 5 other cases of BVAO resulting from GCA and 110 cases with underlying arteriosclerotic disease. Our 3 patients (mean age, 66 yr; range, 60-78 yr) with BVAO resulting from GCA all had initial severe headache followed by the onset of stepwise progressive, partly side-alternating neurologic deficits due to bilateral infarctions in the vertebrobasilar circulation territory. This course, more accelerated in BVAO due to GCA than in BVAO of arteriosclerotic origin, seems to be a typical, if not particular, clinical syndrome. BVAO was the first clinical manifestation of GCA in 1 of our patients and in 1 published case. From a clinical view, BVAO resulting from GCA differs from BVAO of arteriosclerotic origin by the much higher mortality rate (75% versus 19%, respectively), the presence of headache (100% versus 22%), fever (50% versus 0%), and elevated erythrocyte sedimentation rate (ESR in all GCA cases >45 mm/h; no data in the arteriosclerotic patient group), but not by the neurologic signs themselves. Therapy of BVAO resulting from GCA is purely empiric. In view of the serious prognosis, we propose treatment with intravenous high-dose glucocorticoids and additional immunosuppression with cyclophosphamide; the use of anticoagulation depends on the individual patient's estimated risk-benefit profile. Although BVAO due to GCA is rare, physicians and especially rheumatologists or neurologists should be aware of this entity because of its high mortality in patients without immediate introduction of a high-dose immunosuppressive therapy. Suspicion of GCA should arise in a patient aged over 50 years with no other vascular risk factors suffering from bilateral symptoms of ischemia in the vertebrobasilar territory, with a quickly progressing stepwise course and with headache, fever, or history of myalgia. ESR and temporal artery biopsy should be performed without delay. Early diagnosis of GCA is necessary for immediate initiation of intensive antiinflammatory and immunosuppressive treatment, without which progressive deterioration and systemic involvement are likely to be fatal. PMID- 12544707 TI - Audiovestibular manifestations in giant cell arteritis: a prospective study. AB - Giant cell arteritis (GCA) is a multisystemic vasculitis of elderly people that involves large and medium-sized blood vessels with predisposition to the cranial arteries. Some cranial ischemic manifestations, in particular permanent visual loss, have been widely described. Audiovestibular manifestations have been less commonly reported. In the present study we assessed the frequency and outcome of audiovestibular manifestations in a series of GCA and isolated polymyalgia rheumatica (PMR) patients examined prospectively between June 1999 and May 2001 at the single hospital for a defined population. Patients were included in the study if a temporal artery biopsy had been performed and they were examined within a week after beginning corticosteroid treatment. Patients with abnormal otoscopy or tympanogram, history of cerebrovascular complications, syphilis, Meniere and other vestibular syndromes, infections involving the inner ear, barotrauma, or being treated with ototoxic drugs were excluded. During the study period 44 patients with GCA and 10 patients with biopsy-negative isolated PMR were examined. Patients with isolated PMR were younger. Audiovestibular dysfunction was significantly more frequent in GCA patients than in those with isolated PMR and matched controls. Almost 90% of the GCA patients had vestibular dysfunction, which was generally reversible after several days of steroid treatment; after 3 months of treatment, vestibular dysfunction was observed in only 13 (29.6%) of the 44 GCA patients. These patients with persistent vestibular dysfunction were more likely to have persistent head-shaking nystagmus. Twelve (27.3%) of the 44 GCA patients had hearing improvement after 3 months of therapy. After 6 months of therapy, only 1 of the 44 GCA patients had abnormal vestibular tests. However, no additional improvement in hearing function was observed. The present study confirms a high frequency of audiovestibular manifestations in GCA. It also suggests that audiovestibular damage may be reversible in some patients with GCA. PMID- 12544708 TI - High-dose plasma infusion versus plasma exchange as early treatment of thrombotic thrombocytopenic purpura/hemolytic-uremic syndrome. AB - Thrombotic thrombocytopenic purpura and adult hemolytic-uremic syndrome (TTP/HUS) have a substantial mortality rate even with currently available treatments. Although therapeutic plasma exchange is the recommended treatment of TTP/HUS, this cumbersome procedure may not be available for all patients in an emergency. In this context, plasma infusion may represent an alternative first-line therapy. We compared the effectiveness of high-dose plasma infusion (25-30 mL/kg per day) and therapeutic plasma exchange as first-line treatment of adult TTP/HUS at a single center. Two groups of patients with TTP/HUS were identified according to their initial therapy, that is, high-dose plasma infusion (19 patients) and therapeutic plasma exchange (18 patients). Clinical charts and outcomes were retrospectively analyzed. Endpoints for comparison were the duration of platelet counts below 150 x 10 /L and LDH levels above normal values; the volumes of plasma administered and the duration of treatment; complete remission, relapse, and mortality rates; and treatment-related complications. Patients of the 2 groups had comparable clinical and laboratory features on admission. Sixteen patients achieved complete remission in each group. Median times to recovery of platelet counts and LDH levels were comparable between the 2 groups. Eight patients in the high-dose plasma infusion (HD-PI) group were switched to therapeutic plasma exchange because of fluid overload (6 patients), persistent biologic disturbances (1 patient), or unresponsiveness to high-dose plasma infusion treatment (1 patient). This latter patient had severe TTP/HUS that remained refractory to therapeutic plasma exchange and vincristine, and rapidly died. All 7 remaining patients achieved complete remission with therapeutic plasma exchange. Four patients in the HD-PI group and 3 patients in the therapeutic plasma exchange (TPE) group died. In the HD-PI group, 5 patients experienced a transient nephrotic-range proteinuria during treatment. Main complications in the TPE group were collapse (1 patient) and central venous catheter infection (2 patients) or thrombosis (1 patient). Three patients in each group relapsed. High-dose plasma infusion may be an efficient treatment of TTP/HUS in patients who cannot have early plasma exchange. However, the large volumes of plasma required to reach complete remission may result in fluid overload, which may necessitate subsequent therapeutic plasma exchange. PMID- 12544709 TI - The Changing Clinical Spectrum of Human Immunodeficiency Virus (HIV)-Related Oral Lesions in 1,000 Consecutive Patients: A 12-Year Study in a Referral Center in Mexico. AB - In developing countries, the variations in the clinical spectrum of human immunodeficiency virus (HIV)-related oral lesions over time, and the possible effects of antiretroviral therapy, have not been described. In this study we evaluate the clinical spectrum of oral lesions in a series of HIV-infected patients when first examined at the acquired immunodeficiency syndrome (AIDS) clinic of a tertiary care institution in Mexico City, Mexico, and the changes observed over 12 years. All HIV-infected adult patients had an oral examination performed by specialists in oral pathology and medicine who used established clinical diagnostic criteria for oral lesions. Four periods were defined according to the evolving pattern of antiretroviral use: the first 2 were before the introduction of highly active antiretroviral therapy (HAART) and the last 2 were during more established use of HAART. For the statistical analysis the chi square test for contingency tables and the chi-square test for trend were utilized. For dimensional variables, except age, the Kruskal-Wallis or Mann Whitney rank sum tests were used when applicable and trend was tested with the Spearman correlation coefficient. Age was tested through analysis of variance (ANOVA) and linear regression analysis. Alpha value was set at p = 0.05 for each test. In the 12-year study, 1,000 HIV-infected patients were included (87.9% male). At the baseline examination, oral lesions strongly associated with HIV were present in 47.1% of HIV-infected patients. Oral candidosis (31.6%), hairy leukoplakia (22.6%), erythematous candidosis (21.0%), and pseudomembranous candidosis (15.8%) were the most frequent lesions. Oral Kaposi sarcoma (2.3%), HIV-associated periodontal disease (1.7%), and oral non-Hodgkin lymphoma (0.1%) were less frequent. HIV-related oral lesions decreased systematically-by half during the course of the 4 study periods (p < 0.001). Except for Kaposi sarcoma, all oral lesions strongly associated with HIV showed a trend to decrease significantly during the study period. No apparent variation in the occurrence of salivary gland disease or human papillomavirus-associated oral lesions was found. A significant trend to a lower prevalence was observed in the group of patients who were already taking antiretroviral therapy, non-HAART and HAART (p < 0.001 and p = 0.004, respectively). Only a discrete reduction, barely significant, was noted among untreated patients (p = 0.060). By Period IV (1999-2001), those who received HAART showed the lowest prevalence of oral lesions strongly associated with HIV (p < 0.001). Patients with oral lesions strongly associated with HIV had significantly lower median CD4+ counts and higher viral loads than those without oral lesions strongly associated with HIV (p < 0.001 and p = 0.005, respectively). When CD4+ counts were correlated with prevalence of oral candidosis, a consistently negative association was found; this association prevailed even after the study group was partitioned according to period. In this selected cohort of 1,000 patients with HIV infection, the clinical spectrum of HIV-related oral lesions has changed over the 12-year study, with a decreased prevalence of most oral lesions. Our findings probably represent improvements in medical care of HIV-infected persons, earlier detection of HIV-infected patients at the AIDS clinic, the increasing use of prophylactic drugs to prevent secondary AIDS-related opportunistic infections, and, perhaps most important, the availability of potent antiretroviral therapy in recent years, since the introduction of HAART. PMID- 12544710 TI - The AESOP (adenopathy and extensive skin patch overlying a plasmacytoma) syndrome: report of 4 cases of a new syndrome revealing POEMS (polyneuropathy, organomegaly, endocrinopathy, monoclonal protein, and skin changes) syndrome at a curable stage. AB - We describe an easily recognizable and previously not individualized clinical syndrome that can reveal solitary plasmacytoma of bone. We report 4 patients with a slowly extending violaceous skin patch overlying a solitary plasmacytoma of bone, associated with enlarged regional lymph nodes. Biopsies of the cutaneous lesion and the lymph nodes were not specific, although increased dermal mucin deposition and vascular proliferation were present in all skin specimens. Three patients had associated polyneuropathy. One patient had POEMS (Polyneuropathy, Organomegaly, Endocrinopathy, Monoclonal protein, and Skin changes) syndrome at the time the plasmacytoma was diagnosed. Another patient developed POEMS syndrome, from which he died, 4 years after excision of the plasmacytoma. The 3 other patients were treated either with irradiation or with a combination of irradiation and surgery, and recovered completely, including from the associated neuropathy and/or POEMS syndrome. We suggest calling this unique and distinctive clinical presentation the for PMID- 12544711 TI - The long-term mortality and morbidity of Behcet syndrome: a 2-decade outcome survey of 387 patients followed at a dedicated center. AB - We surveyed the 20-year outcome of a cohort of patients with Behcet syndrome with emphasis on both mortality and morbidity. During 1999 and 2000, we collected outcome information on 387/428 (90.4%) of a cohort of patients (262 male, 125 female) who had registered in a dedicated outpatient clinic between July 1977 and December 1983. In 245/345 (71.0%) patients, outcome information was based on a formal hospital reevaluation, and in the remaining patients, on detailed telephone interviews. Forty-two patients (9.8%) (39 male, 3 female) had died, mainly due to major vessel disease and neurologic involvement. Mortality, as measured by standardized mortality ratios (SMR), was specifically increased among young males, among whom morbidity was also the highest. However, the SMR tended to decrease significantly with the passage of time. The same was also true for all mucocutaneous and articular manifestations. Both the onset of eye disease and its greatest damage were also usually within the first few years of disease onset. These suggest that the "disease burden" of Behcet syndrome is usually confined to the early years of its course, and in many patients the syndrome "burns out." However, central nervous system involvement and major vessel disease are exceptions. They can have their onset late (5-10 yr) during the disease course. As reflected in the mortality figures, the disease was less severe among the females for almost each disease manifestation. There were no female patients with arterial aneurysms. Severely impaired vision did not always mean an eventual loss of useful vision, and those patients with a late onset of eye disease had a better visual prognosis. PMID- 12544713 TI - Imaging of arteries. PMID- 12544714 TI - Assessment of large artery function. PMID- 12544715 TI - Neurohormonal assessment of cardiac function. PMID- 12544716 TI - Kawasaki disease--from a mystery to a paradigm. PMID- 12544717 TI - Endogenous heparin--a protective marker in patients with myocardial infarction. AB - BACKGROUND: In patients with myocardial infarction and initial high serum levels of immunoglobulin E (IgE), lower mortality and a smaller quantity of complications were observed. The aim of this study was to evaluate whether there was a relationship between IgE serum levels and plasma concentration of endogenous heparin in patients with myocardial infarction. Furthermore, we attempted to analyse whether there was relationship between endogenous heparin plasma concentrations and the specific markers (thrombin-anti-thrombin ?TAT?complexes) for thrombinogenesis. METHODS: Thirty-five healthy individuals and 64 patients with recent myocardial infarction (MI), aged 37-70 years were enrolled in the study. Patients with MI who had suffered cardiac arrest and cardiogenic shock before admission to hospital, or had inflammatory diseases or a negative allergic history were excluded. Serum IgE concentrations (UniCAP), TAT complexes in plasma (ELISA, Behring) and endogenous heparin plasma levels (Coatest Heparin) were measured at the time of admission and before treatment. RESULTS: There were striking differences between the heparin concentrations in patients with MI and the control group ( p< 0.0001). Higher levels of IgE were strongly correlated with higher concentrations of endogenous heparin ( r= 0.80, p < 0.001). Strong statistically significant negative correlations between higher endogenous heparin concentrations and TAT complex levels were also found (r = 0.57, p < 0.001). CONCLUSION: The results suggest that high concentrations of endogenous heparin in association with high IgE concentrations and their negative influence on thrombinogenesis, may act as a marker for a favourable prognosis in patients with MI. PMID- 12544718 TI - Efficacy of monotherapy compared with combined antianginal drugs in the treatment of chronic stable angina pectoris: a meta-analysis. AB - OBJECTIVE: To determine the relative efficacy of antianginal drugs administered as monotherapy or in combination in patients with chronic stable angina. METHODS: A meta-analysis was performed on randomized trials, published in English between 1980 and 1999, that directly compared combined treatment and monotherapy. Twenty two articles were included, all on the comparison of -blocker monotherapies to their combination with a calcium antagonist and 10 on the comparison of calcium antagonist monotherapies to their combination with a -blocker. RESULTS: Time to 1 mm ST-segment depression, total exercise duration and time to onset of anginal pain were significantly increased with the combined therapy compared to -blocker alone (by 8, 5 and 12%, respectively). Only time to 1 mm ST-segment depression was significantly increased with the combined therapy compared to calcium antagonist alone (by 9%). For all these parameters, the adjusted differences were significant only within 6 h following drug intake and were not significant after 6 h. No analysis of safety data could be performed. CONCLUSION: As far as exercise testing is concerned, the combination of a calcium antagonist and a blocker is statistically more effective than either monotherapy. Further studies are needed to confirm the higher efficacy after the first 6 h following drug intake. PMID- 12544719 TI - The role of coronary bypass operation on children with Kawasaki disease. AB - BACKGROUND: Kawasaki disease, initially called mucocutaneous lymph node syndrome was reported 35 years ago as a new inflammatory disease in infants and children and is characterized by a variety of symptoms and signs resulted from systemic vasculitis. Although the etiology of the disease remains unknown, its serious coronary complications have been proved to cause ischemic heart disease in children, and are now the most common cause of pediatric coronary disease in the world. The incidence of serious coronary sequelae is fortunately low (2-3% of patients with Kawasaki disease), but once myocardial infarction occurs in children, the mortality is quite high (22% at the first infarction). Development of surgical treatment for the disease was essential in preventing premature death and improving the quality of life of children. METHODS AND RESULTS: Coronary revascularization surgery was attempted following careful evaluation of characteristic patters of coronary aneurysms and obstructions secondary to Kawasaki disease, although the surgical efficacy was initially questioned because the disease is inflammatory vasculitis in origin. The operation utilizing the pedicled internal thoracic artery has been demonstrated quite successful and now established as a reliable treatment for inflammatory coronary obstructions due to Kawasaki disease (the Kitamura Operation). There is valid evidence for the internal thoracic artery graft being a viable structure, accommodating in length and diameter for the growth of children. Results of the surgery and long-term prognosis are favorable and postoperative quality of life is markedly improved. CONCLUSIONS: Coronary bypass operation utilizing the pedicled internal thoracic artery is a safe and reliable surgical modality for coronary artery sequelae in children due to Kawasaki disease. Long-term follow-up results up to 20-years are quite satisfactory. PMID- 12544720 TI - The place of beta 2 glycoprotein 1 in the assessment of antiphospholipid syndrome. AB - beta 2 glycoprotein 1 (beta2GP1) is a phospholipid-binding protein implicated in the development of antiphospholipid antibodies, associated with thromboembolic complications and fetal morbidity and death, and is thought to corrrelate better than anticardiolipin (aCL) assays. We analysed the role of beta2GP1 in assessing 86 patients being investigated for antiphospholipid syndrome. Thirty-nine patients had 3 tests: [lupus anticoagulant (LA), aCL and beta2GP1], and a further 46 had aCL and beta2GP1. Sixty-one patients had completely negative tests. Five patients had beta2GP1 as the only positive result. 80% of this group had recurrent miscarriage suggesting that beta2GP1 may be an useful adjunct to aCL and LA testing in patients with a significant obstetric history. PMID- 12544721 TI - Development of an experimental model of pre-thrombosis in rats based on Wessler's principle using a calibrated venous stasis. AB - We have developed a model of a pre-thrombotic state in rats based on venous stasis induced by partial ligature of the inferior vena cava. The degree of stenosis was calibrated by using variations in upstream venous pressure. Different degrees of stasis were tested in order to obtain a pre-thrombotic state. Increasing doses of thromboplastin were infused. The thrombogenic potential of this model was evaluated by measuring thrombus weight and by the increase in levels of thrombin-antithrombin complexes. A pre-thrombotic state was induced by 2 h of exposure to a 40% stasis obtained by increasing by 40% the upstream venous pressure (mean thrombus weight, 0.2 +/- 0.6 mg). In these conditions of stasis, low doses of thromboplastin induced venous thrombosis (mean weight, 23 +/- 20 mg; P < 0.05). The increase in thrombus size was correlated to the rise in thrombin-antithrombin levels (r = 0.53, P < 0.001). In conclusion, we have developed the first animal model in which venous stasis can be calibrated by varying the degree of stenosis of the inferior vena cava. This model could be used to study the kinetics of biological markers of hypercoagulability, to study the pathogeny of thrombosis or to evaluate the therapeutic efficacy of new drugs in pre-clinical trials. PMID- 12544722 TI - The initial phase of oral anticoagulation with warfarin in outpatients with deep venous thrombosis. AB - Patients with deep venous thrombosis (DVT) treated out of hospital usually start warfarin with the recommended 5 mg loading dose and have their International Normalized Ratio (INR) test performed every 2-3 days. Thus, achievement of the therapeutic range may be more difficult than for inpatients, possibly resulting in extended duration of low molecular weight heparin (LMWH) treatment. We retrospectively examined the charts of 55 DVT outpatients (mean age, 61.4 years; 30 males) to assess the actual duration of LMWH treatment and to identify predictors of a slow achievement of the INR range. Thirty patients (54.4%) reached the therapeutic INR range and stopped LMWH within 7 days, and 25 patients (45.6%) had to continue for an average of 10.5 days. The latter group was significantly younger than the former (57 and 65 years, respectively; P = 0.039). Patients younger than 60 years old had an odds ratio for an extended treatment of 4.92 (P = 0.0057). Algorithms with different loading doses of warfarin according to age should be proposed for outpatient treatment of DVT. PMID- 12544723 TI - Viral load and disease progression as responsible for endothelial activation and/or injury in human immunodeficiency virus-1-infected patients. AB - The endothelium participates in haemostasis, inflammation, blood pressure regulation and other physiological systems. Consequently, endothelial dysfunction has been related to hypertension, thrombosis and atherosclerosis. Both von Willebrand factor (vWF) and tissue-type plasminogen activator (t-PA) are synthesized by the endothelium and their plasma levels increased during endothelium activation or injury. So far, they are well-known markers of endothelial cell function. Many circumstances activate or damage the endothelium, such as viruses, bacterium and inflammation. Circulating vWF and t-PA were studied in 92 unselected human immunodeficiency virus-1 (HIV-1)-infected patients [27 patients with and 65 patients without acquired immunodeficiency syndrome (AIDS)] and correlated with plasma levels of pro-inflammatory cytokines (tumour necrosis factor-alpha, interleukin-6), viral load, CD4 T-cell count and infectious status. HIV-1-infected patients had significantly higher plasma levels of vWF (152 versus 90%), tumour necrosis factor-alpha (31.3 versus 9.0 pg/ml) and interleukin-6 (3.5 versus 1.9 pg/ml) but not t-PA (5.9 versus 4.2 ng/ml) than the control group. These two endothelial markers correlated significantly with viral load and interleukin-6 levels in HIV-1-infected patients. The highest levels of vWF and t-PA were found in patients with AIDS. In conclusion, endothelial cell perturbation is present in HIV infection and may be a consequence of different mechanisms such as viral load, cytokines and advanced diseases. PMID- 12544724 TI - Reactive hyperemia and tissue-type plasminogen activator release in hypertensive men. AB - A relationship may exist between endothelial-mediated vasodilation and tissue type plasminogen activator (t-PA) release. However, the existing evidence is mainly based upon exogenous agonist administration, and needs testing under more physiological conditions. We evaluated the link between t-PA, the key fibrinolytic factor in man, and forearm reactive hyperemia, a model of endogenous endothelial-mediated vasodilation, in 13 uncomplicated hypertensive subjects and six elderly hypertensive patients with atherosclerotic peripheral vascular disease and hypercholesterolemia (i.e a group in whom post-ischemic hyperemia was probably defective because of dysfunctional endothelium). To characterize further the phenomenon, 29 additional uncomplicated hypertensive patients underwent intra arterial drug infusions. Study variables were forearm blood flow (strain-gauge plethysmography), arterial and venous concentrations of t-PA mass concentrations, and calculated net release (forearm plasma flow x veno-arterial differences). Reactive hyperemia was induced by inflating a cuff midway between systolic and diastolic pressure for 10 min; blood and forearm blood flow were sampled before and after cuff release. Post-ischemic t-PA release increased in uncomplicated hypertensives, and did not change in hypercholesterolemic atherosclerotic patients in whom post-ischemic vasodilation was negligible. Local adenosine (n = 9), acetylcholine (n = 12) and bradykinin (n = 8) vasodilated similarly, but only bradykinin increased t-PA release. Thus, reactive hyperemia stimulates t-PA release, and that relationship is altered when endothelium is dysfunctional. Release of t-PA is independent of forearm vasodilatation, adenosine or biological products of muscarinic stimulation and may, perhaps, be related to the activity of the endogenous kininogen/kinin system. PMID- 12544725 TI - Safety of dental extraction among consecutive patients on oral anticoagulant treatment managed using a specific dental management protocol. AB - We found no prospective studies on dental extraction in anticoagulated patients in the literature, even though most authors suggest that there is no need to change anticoagulant treatment and to utilize a local haemostatic measure after extraction. In the present study, we have verified the incidence of bleeding complications after dental extraction in a group of 250 consecutive anticoagulated patients. Two hundred and fifty non-anticoagulated subjects requiring dental extraction represented the control group. In all patients, anticoagulant treatment was not changed (International Normalized Ratio, 1.8-4) and local haemostatic measures (fibrin sponge, silk suture and a gauze saturated with tranexamic acid) were used. All procedures were performed in an outpatient clinic setting. We registered four bleeding complications in the group of anticoagulated patients and three in the control group. The difference of bleeding complications in the two groups was not statistically different (relative risk, 1.14; 95% confidence interval, 0.29-6.04; P = 0.7). None of the post-operative late bleeding required hospitalization and/or blood transfusions, and further local measures were sufficient to stop the bleeding. The protocol proposed in the present study makes dental extractions in anticoagulated patients possible on an outpatient basis with a cost reduction for the community and minor discomfort for the patients. PMID- 12544726 TI - Gorog Thrombosis Test: a global in-vitro test of platelet function and thrombolysis. AB - This is the first laboratory evaluation of a new instrument, designed to test both platelet function and thrombolytic activity from a native blood sample, in vitro. The inventor assumed that the reduction and arrest of blood flow was due to activation, aggregation and stabilized thrombus formation by shear-activated platelets, and that re-establishment of flow was due to thrombolysis. Morphologic and functional studies presented here confirm these mechanisms. In vitro tests provided incontestable evidence for the principal role of platelets in the obstruction of flow (occlusion time) and for thrombolysis as the principal mechanism underlying the restoration of blood flow (lysis time). In addition to aggregation, it is the explosive generation of thrombin by shear-activated platelets that results in the formation of an occlusive haemostatic thrombus. Anticoagulation of blood completely prevented occlusion. Platelet-rich thrombus formation (occlusion time) was dose-dependently inhibited by monoclonal antibody against platelet glycoprotein (GP) Ib (6B4 and 12E4), aurin tricarboxylic acid, monoclonal antibody against platelet GPIIb/IIIa (MA-16N7C2 and abciximab), a synthetic GPIIb/IIIa antagonist (TAK-029), thrombin inhibitor (argatroban), and anti-von Willebrand factor, but not by anti-fibrinogen. Plasminogen activator streptokinase (Varidase) and tissue-type plasminogen activator (Monteplase) dose dependently enhanced thrombolysis (lysis time) without affecting platelet function (occlusion time). The test is specific for thrombolysis. The plasmin inhibitor tranexamic acid prevented plasminogen activator-induced thrombolysis, while inhibition of clot retraction by cytochalasin B did not affect the lysis time. This rapid and sensitive global test of platelet function and thrombolytic activity could be of great value both in research and in clinical practice. PMID- 12544727 TI - Ex-vivo regulation of endotoxin-induced tissue factor in whole blood by eicosanoids. AB - The influence of several eicosanoids of the lipoxygenase pathway was examined in an ex vivo system of human whole blood subjected to stimulation by lipopolysaccharide (LPS). Exogenously added leukotriene B4 [5(S),12(R)-dihydroxy 6,14-cis-8,10-trans-eicosatetraenoic acid (LTB4)] or 12(S) hydroxyeicosatetraenoic acid (12(S)-HETE) significantly (P<0.05) enhanced LPS evoked expression of monocyte tissue factor (TF) activity in a concentration dependent manner. 15(S)-HETE, on the other hand, exerted such activity only when added at certain concentrations, whereas 5(S)-HETE was devoid of any apparent activity. LPS-induced TF activity was inhibited by the lipoxygenase inhibitors nordihydroguaiaretic acid, CGS 23885 and ZM 230487, by 59, 32 and 88%, respectively. Furthermore, the production of LTB4 in LPS-stimulated whole blood was investigated, in the absence or presence of either tumor necrosis factor alpha (TNFalpha) or phorbol-12-myristate-13-acetate (PMA). LPS alone induced a moderate time-dependent and concentration-dependent release of LTB4, reaching the maximum concentration (1260 +/- 202 pg/ml) within 90 min at 5 ng/ml LPS. The prior and concurrent presence of PMA (5 ng/ml) or TNFalpha (10 ng/ml) further enhanced the LTB4 production approximately twofold (P < 0.05). TNFalpha added alone evoked approximately twice the LTB4 production seen when LPS (2200 +/- 243 versus 1260 +/- 203 pg/ml) was added alone. Considering these results, LPS and TNFalpha emerge as important agonists of LTB4 production in whole blood. LTB4 in turn appears to be of importance for the expression of TF in monocytes, potentially amplifying the thrombogenic potential of these cells. PMID- 12544728 TI - Coagulation factor V G allele and HR2 haplotype: factor V activity, activated protein C resistance and risk of venous thrombosis. AB - Factor V Leiden is a well-known risk factor for venous thrombosis. The dual role of factor V as a coagulatory and anticoagulatory cofactor permits the assumption that further mutations in the factor V gene are of importance in the study of the risk of thrombosis. Relevant studies to date have given rise to a controversy over this risk for the HR2 haplotype. For the G allele, defined in our work group as a G at the nucleotide positions 2391, 2663, 2684 and 2863, there have been to date no other investigations of thrombotic risk. In a case-control study on 347 patients with deep venous thrombosis (DVT) and 282 controls, we investigated the association of the HR2 haplotype and the G allele with DVT. We found no association between HR2 haplotype and DVT [odds ratio (OR) 0.87; 95% confidence interval (CI) 0.58-1.30; P = 0.537]. The frequency of the G allele was, on the contrary, higher in the control group than among the patients (OR 0.68, 95% CI: 0.53 to 0.89; P = 0.005). The factor V activity of the HR2 carriers was lower than that of the wild type and G allele carriers. The HR2 haplotype exhibited a moderate influence on activated protein C response. This study presented no evidence of thrombotic risk for the HR2 haplotype alone. The results here permit the assumption of a protective effect of the G allele. The source of a possible protective influence of the G allele on thrombotic risk is at present unclear. PMID- 12544729 TI - Antiphospholipid antibodies in young women with and without oral contraceptive use. AB - The role of thrombophilia in the elevated risk of thromboembolism during oral contraceptive use has been established. We performed a cross-sectional study among young women to survey the occurrence of antiphospholipid antibodies among users and non-users of oral contraceptives. Serum levels of immunoglobulin (Ig)G, IgA and IgM isotypes of anti-beta2-glycoprotein I and anticardiolipin antibodies were measured by validated enzyme-linked immunosorbent assay methods. Combining all types of antiphospholipid antibodies, pill-users had an elevated antibody titre more than twice as frequently as non-users (odds ratio, 2.3; 95% confidence interval, 1.1-5.1). The higher frequency of elevated antibody titre was related most commonly to IgG type anti-beta2-glycoprotein I antibodies. Oral contraceptive use increases the risk of elevated antiphospholipid antibody levels among asymptomatic young women. PMID- 12544730 TI - Idraparinux and liver enzymes: observations from the PERSIST trial. AB - A potential influence of idraparinux--a synthetic analogue of the pentasaccharide sequence in heparins--on plasma liver enzyme levels was analysed in 37 patients suffering from deep vein thrombosis and participating in the PERSIST trial. Plasma gamma-glutamyl-transferase, aspartate aminotransferase and alanine aminotransferase were determined prior to enoxaparin treatment (screening), prior to randomization (baseline) and once weekly during the 12-week treatment period. Patients were initially treated with weight-adjusted enoxaparin for 4-7 days and then randomized to either idraparinux (2.5, 5, 7.5 or 10 mg) or warfarin. Gamma glutamyl-transferase was significantly increased after administration of enoxaparin at the baseline visit (P = 0.004) and in week 2 (P = 0.009) to return to screening levels in week 3 for the remaining study period (all P > 0.05). Aspartate aminotransferase (P = 0.001) and alanine aminotransferase (P < 0.001) were significantly increased at the baseline visit and returned to screening values at week 2 for the remaining study period (all P > 0.05). There was no significant difference between the mean values of plasma liver enzymes of the four idraparinux groups and the warfarin group in all 13 measurements. We concluded that idraparinux in contrast to enoxaparin does not increase plasma liver enzymes significantly. PMID- 12544731 TI - Pre-thrombotic state and impaired fibrinolytic potential in coronary heart disease patients with left ventricular dysfunction. AB - Patients with coronary heart disease (CHD) are at considerable risk for recurrent ischaemic events. A pre-thrombotic state and/or impaired fibrinolysis might play an important role in causing recurrent ischaemic events. Two hundred and fifty seven CHD patients underwent the dobutamine stress echocardiography test (DSE) to investigate the possible presence of inducible ischaemia; 89 patients showed evidence of stunned and/or necrotic myocardium (resting wall motion abnormalities). Factor VIII activity and fibrinogen levels were significantly higher in patients with stunned/necrotic myocardium than in CHD patients with normal resting wall motions (factor VIII activity, P = 0.004; fibrinogen, P = 0.04). Of interest, after stimulating the fibrinolytic system with the DSE test, plasminogen activator inhibitor-1 activity was significantly higher in patients with necrotic/stunned myocardium than in patients with resting normal wall motion (P = 0.03), whereas tissue-type plasminogen activator activity after the DSE test was significantly lower in patients with stunned/necrotic myocardium than in patients with normal wall motion (P = 0.001). Overall, 30 CHD patients developed induced ischaemia (new wall motion abnormalities) during the DSE test. CHD patients with stunned and/or necrotic myocardium presented decreased fibrinolytic potential and the presence of a hypercoagulable state due to increased factor VIII activity, and fibrinogen levels. Therefore, these CHD patients must be considered at high risk of re-developing coronary thrombosis and might benefit from a more aggressive anticoagulant therapy. PMID- 12544732 TI - Thrombotic thrombocytopenic purpura-hemolytic uremic syndrome after bupropion treatment for smoking cessation. AB - Bupropion hydrochloride is an effective drug for people who want stop smoking, and its use has recently increased in many countries. The main side effects of this drug are related to its dopaminergic activity and are dose dependent. To date, no cases of thrombotic thrombocytopenic purpura-hemolytic uremic syndrome (TTP-HUS) have been reported in literature. Here we describe a case of a woman who developed a sporadic form of TTP-HUS during a treatment with bupropion for smoking cessation, successfully treated with plasma ex-change therapy. The authors wish to make readers aware of bupropion as a possible cause of this potentially lethal disease. PMID- 12544733 TI - Right ventricular mass and pulmonary embolism in a patient with essential thrombocythemia. AB - We report a case of a patient with essential thrombocythosis with a right ventricular mass and massive pulmonary embolism following an abdominal operation and who was operated on due to severe hemodynamic instability. PMID- 12544734 TI - Ischaemic strokes and homozygosity for the alpha2 807T allele of the platelet collagen receptor in young monozygotic twins. AB - A nucleotide 807T variant of the glycoprotein Ia gene that correlates with increased platelet surface levels of the platelet collagen receptor alpha2beta1 was recently found to be associated with an increased risk of ischaemic stroke in younger patients. We report the history of twins who developed ischaemic strokes and were shown to be homozygous for the alpha2 807T allele. The twins developed ischaemic strokes at the ages of 23 and 33 years, one of them with recurrent events. They had no conventional risk factors. Cardiac and vascular investigations were normal and no aetiology could be found. There was a family history of cerebrovascular disease. Genotyping of glycoprotein alpha2 C807 T was performed and both twins were found to be homozygous for the 807T allele. This allele probably contributed to the occurrence of strokes in these young men. Further prospective studies are needed to evaluate whether screening for this polymorphism should be considered in young patients with unexplained stroke, particularly when a positive family history was found. PMID- 12544735 TI - Acquired factor V inhibitor complicated by Hashimoto's thyroditis, primary biliary cirrhosis and membranous nephropathy. AB - A 59-year-old man diagnosed as having Hashimoto's thyroditis, primary biliary cirrhosis (PBC) and membranous nephropathy (MN) showed consciousness disturbance, convulsions of the upper part of his body, and rapid progression of anemia, which seemed to be derived from subdural and retroperitoneal hemorrhage, respectively. He had been diagnosed as having eosinophilia about 6 weeks before the attack. Coagulation tests revealed a prolonged activated partial thromboplastin time and prothrombin time, which could not be normalized by mixing with normal plasma. Factor V (FV) activity was severely decreased and the purified immunoglobulin G of the patient inhibited normal plasma FV activity in a dose-dependent manner, suggesting the presence of antibody-mediated circulating inhibitors specific for FV. Treatment with steroids and azathioprine as well as plasmapheresis led to improvement of his clinical symptoms, normalization of the coagulation tests, and disappearance of eosinophilia. However, the inhibitor reappeared about 7 months later in association with eosinophilia, which was also improved by steroid therapy. To our knowledge, this is the first report of the co-existence of these three kinds of immune-mediated disorders, and the first report concerning the association between acquired FV inhibitors and PBC with MN. A new unknown immune mechanism, which causes eosinophilia, may be involved in the development of the FV inhibitor in this patient. PMID- 12544736 TI - Severe venous thromboembolism in a young man with Klinefelter's syndrome and heterozygosis for both G20210A prothrombin and factor V Leiden mutations. AB - Klinefelter's syndrome is the most common cause of primary testicular failure, resulting in impairment of both spermatogenesis and testosterone production. It is a chromosomal disorder characterized by small, firm testes, azoospermia, gynecomastia, varying degrees of eunuchoidism and testosterone deficiency with elevated gonadotropin plasma levels. In Klinefelter's syndrome there is an increase of certain systemic diseases including venous thromboembolism. An increased thromboembolic risk in hypogonadic men has been explained with hypofibrinolysis due to androgen deficiency. Only two cases have been reported about the association between Klinefelter's syndrome and well-known congenital or acquired thrombophilias. We report the case of a 39-year-old patient with Klinefelter's syndrome who underwent severe deep venous thrombosis with pulmonary embolism, in the absence of any circumstantial triggering event. Further examinations also showed a double heterozygosis for G20210A prothrombin and factor V Leiden mutations. This case suggests that the increased thromboembolic risk, reported in Klinefelter's syndrome, can be worsened by the co-existence of one or more well-known thrombophilic conditions, as shown by the relatively young age of the patient. More studies are needed to clearly understand the pathogenesis of venous thromboembolism in males affected by Klinefelter's syndrome. PMID- 12544737 TI - A differentiated approach to assess the diagnostic usefulness of a rapid particle gel immunoassay for the detection of antibodies against heparin-platelet factor 4 in cardiac surgery patients. AB - Enzyme-linked immunosorbent assay (ELISA) methods have shown to be of a low specificity for confirming heparin-induced thrombocytopenia (HIT) in cardiac surgery patients. In other patient collectives, a new antigen test [ID-HPF4 Particle Gel Immuno Assay (PaGIA); Diamed, Cressier sur Morat, Switzerland] was recently reported to exhibit a better specificity than the ELISA. Since a more specific test would serve as a useful tool when diagnosing HIT, a prospective study was carried out to elucidate the specificity and diagnostic usefulness of PaGIA in cardiac surgical patients. As assessed in cardiac surgery patients without HIT, the proportion of PaGIA-positive samples was lowest at baseline (16%; 13/74), increasing to 31% (19/61) at postoperative days 5-9 (P = 0.06) and 69% (29/42) at postoperative days 10-18 (P < 0.001 versus postoperative days 5 9). The respective proportions in an ELISA (HPF4 ELISA; Stago, Asnieres sur Seine, France) were lower. Because the prevalence of antibody positivity increases during the postoperative course, a differentiated mathematical model was used to assess predictive values and likelihood ratios (LR) of the PaGIA at three different phases of hospitalization. Calculating LR revealed -LR to be 0.07 at baseline, 0.09 during postoperative days 5-9, and 0.19 during postoperative days 10-18, while +LR were 5.9 at baseline, 3.0 during postoperative days 5-9, and 1.4 during postoperative days 10-18. Thus, in cardiac surgery patients, the PaGIA can be regarded as a useful tool in excluding rapid onset and typical onset of HIT, and further provides valuable information in the confirmation of rapid onset of HIT. Using a differentiated approach for calculation of predictive values and LR, the present study demonstrates that PaGIA, despite a lower specificity than the employed ELISA, in some situations is a valuable tool when diagnosing HIT in cardiac surgery patients. PMID- 12544738 TI - A splicing donor site point mutation in intron 6 of the plasmin inhibitor (alpha2 antiplasmin) gene with heterozygous deficiency and a bleeding tendency. AB - A new case of familial plasmin inhibitor (alpha2 antiplasmin) deficiency is reported. The bleeding symptoms are moderate, happening after surgery or trauma or consisting of abnormal uterine bleeding induced by hormone replacement therapy. It is easily corrected with tranexamic acid. Gene sequencing makes it possible to find a splicing donor site mutation of intron 6, leading to exon 6 skipping. Neither a shortened variant nor an abnormal plasmin interaction was found in plasma by immunoblotting, and fibrin binding is unaffected. The mutation is heterozygous, associated with an intermediate decrease of both antiplasmin activity and antigen levels, and was found in four other family members out of five tested. It is different from the five mutations previously reported. At the time of diagnosis, the patient was living in Artas, France, allowing the defect to be named plasmin inhibitor (alpha2 antiplasmin) Artas. PMID- 12544739 TI - Association of anti-heparin platelet factor 4 antibody levels and thrombosis in pediatric intensive care patients without thrombocytopenia. AB - Anti-heparin platelet factor 4 (anti-HPF4) antibodies have been demonstrated to play a pathogenetic role in the development of heparin-induced thrombocytopenia. In adults, the presence of anti-HPF4 antibodies without thrombocytopenia has been reported not to confer a thrombotic risk. To investigate whether this also holds true for children, we performed a case-control study in heparin-exposed patients from a pediatric intensive care unit. During the 30-month study period, 612 patients received heparin for at least 5 days. Of these, 10 patients developed thrombosis without thrombocytopenia and constituted the study group. These patients were compared with 19 matched control patients with neither thrombosis nor thrombocytopenia. Anti-HPF4 antibody levels were measured using an enzyme linked immunosorbent assay (Asserachrom HPF4). All thrombosis patients and controls had lower anti-HPF4 antibody levels than the cut-off level recommended by the manufacturer for adults. However, median anti-HPF4 antibody levels were significantly higher in the thrombosis patients [51% of the manufacturer's cut off; interquartile range (IQR), 47-53%] than in the control group (23%; IQR, 9 36%) (P = 0.004). At an anti-HPF4 cut-off level of 45%, the odds ratio for a thrombotic event amounted to 34 (95% confidence interval, 4.4-261.8), indicating an association between anti-HPF4 antibody levels and thrombosis despite the absence of thrombocytopenia. A role of anti-HPF4 antibodies in the development of catheter-related thrombosis is suggested. PMID- 12544740 TI - Recombinant factor VIIa in preterm neonates with prolonged prothrombin time. AB - The purpose of the study was to estimate the effects of a minimal and a normal dose of rFVIIa on prothrombin time, and to compare it to the effect of a standard volume of fresh frozen plasma. Recombinant factor VIIa (rFVIIa) was tested in babies with gestational age less than 33 weeks and prothrombin time greater than 48 s (reference 23.7 s; i.e. inclusion at international normalized ratio 2.0) during the first 7 days of life. Firstly, 5 microg/kg rFVIIa was given to five babies. One hour after the bolus, prothrombin time had significantly shortened from a mean of 64 s (range 53-79 s) to 46 s (range 39-56 s). Next, the effect of 80 microg/kg rFVIIa was compared to the effect of 10 mL/kg fresh frozen plasma. Prothrombin time at inclusion was 68 s (range 53-92 s). One hour after treatment, prothrombin time was 33 s (range 21-42 s) in the six babies given rFVIIa, compared with 51 s (range 40-59 s) in four babies given fresh frozen plasma (P = 0.02). Prothrombin time remained lower in the rFVIIa group (P = 0.01) at 3 h than in the frozen plasma group, whereas at 6 and 12 h the differences were statistically insignificant. One baby continued to have slight bleeding from the umbilical site and one baby developed severe pulmonary haemorrhage in spite of rFVIIa treatment. In conclusion, rFVIIa partially normalized prothrombin time in preterm babies. There was no evidence of side effects. Future randomized studies are needed to evaluate the haemostatic effect. PMID- 12544741 TI - Idiopathic interstitial pneumonias: an update. PMID- 12544742 TI - Detection of lung perfusion abnormalities using computed tomography in a porcine model of pulmonary embolism. AB - The purpose of this study was to identify perfusion defects of the lung using computed tomography (CT). A balloon catheter was placed in a lobar pulmonary artery of six anesthetized, ventilated, juvenile pigs to simulate occlusive segmental embolus. Contrast medium was injected via a central venous catheter at rates of 1.5, 3, 4.5, and 9 ml/s in each pig. A 40-second single-level cine CT was acquired distal to the inflated balloon during suspended inspiration. Three computer-manipulated images (time to maximal enhancement, change in maximal attenuation, maximal contrast minus precontrast subtraction) were generated using custom software and compared with the unmodified maximal enhancement and precontrast images. Two independent observers identified perfusion defects and scored the level of confidence (5-point scale) on all five images. Regions of interest were drawn in perfused and nonperfused lung and time-attenuation curves were generated. Perfusion defects were accurately (99.8 +/- 0.3%) and confidently (4.5 +/- 0.6) detected and there was excellent interobserver agreement (Kappa 0.99 +/- 0.02) on all computer-manipulated images. There was a significant increase in confidence (p < 0.05) between contrast medium injection rates of 1.5 and 9 ml/s. A linear relationship exists (r = 0.88) between injection rate and change in maximal attenuation. In conclusion, perfusion defects of the lung are seen using computer-manipulated CT images. PMID- 12544743 TI - Usefulness of computed tomography-guided transthoracic small-bore coaxial core biopsy in the presence of a pneumothorax. AB - Transthoracic needle biopsy (TNB) is usually avoided in the presence of pneumothorax. The authors performed computed tomography (CT)-guided transthoracic core biopsy in the presence of pneumothorax in 13 patients (4.9%) selected from 265 patients who received CT-guided TNB over 4 years. These iatrogenic pneumothoraces were induced by previous ultrasound (US)-guided TNB (n = 5), transbronchial lung biopsy (n = 4), and CT-guided biopsy (n = 4). The time interval between previous thoracic intervention and CT-guided TNB ranged from 0 hours to 9 days after transbronchial lung biopsy (average, 4 days). A diagnostic core biopsy was performed in 12 of the 13 patients. Seven lesions proved to be malignant and five were benign. Failure of CT-guided transthoracic core biopsy occurred in a single patient with a previous US-guided biopsy within 24 hours. This patients demonstrated a progressively enlarging pneumothorax and was treated with air aspiration with CT guidance. A successful second biopsy was performed 7 days later after full expansion of the lung. There were no complications related to the procedures. The authors' experience suggests that CT-guided transthoracic core biopsy using small-bore coaxial technique can be safely performed with high diagnostic yield in patients with stable iatrogenic pneumothorax. PMID- 12544744 TI - Radiographic relationship of the origin of the bronchial arteries to the left main bronchus. AB - The purpose of this study was to demonstrate that the relationship of the left main bronchus to the bronchial arteries is a reliable radiographic landmark for accurate catheterization of the right and left bronchial arteries. Sixty-six bronchial angiograms and chest radiographs in 43 patients were reviewed. The relationship of the tip of the catheter during initial bronchial angiography (used as the point of bronchial arterial origin) and the radiolucent shadow of the left main bronchus in patients with and without radiographic evidence of anatomic distortion caused by pulmonary disease was categorized into three groups: within the shadow of the left main bronchus, near the left main bronchus, and far from the left main bronchus. The origin of the right and left bronchial arteries was within or near the shadow of the left main bronchus in 94% of bronchial angiograms performed in patients with and without distorted thoracic anatomy. The most frequent location was within the shadow of the left main bronchus. The left main bronchus can be used as a reliable reference point for catheterization of the right or left bronchial arteries in patients with and without thoracic anatomic distortion. PMID- 12544745 TI - Computed tomography features of the thoracic manifestations of Wegener granulomatosis. PMID- 12544746 TI - Gunshot wound to the thorax with bullet embolization to the external carotid artery. AB - Gunshot wounds are associated with major morbidity and mortality rates. If the patient survives the initial insult, all gunshot wounds require an assessment of the organs at risk and localization of the projectile. This case of a gunshot wound to the thorax with bullet embolization from the left ventricle to the right external carotid artery provides an overview of the imaging in this clinical scenario. Typically, right cardiac perforation results in embolization into the pulmonary vasculature, whereas left cardiac perforation results in embolization into the extremities. PMID- 12544747 TI - Levitating consolidation in eosinophilic lung disease. AB - The authors describe a case of histologically proven eosinophilic lung disease in a patient with ulcerative colitis. The patient was not using sulfasalazine or other medications known to be associated with lung disease. Serial chest radiographs revealed an unusual pattern of levitating consolidation. PMID- 12544748 TI - High-resolution computed tomography appearance of pulmonary Mycobacterium avium complex infection after exposure to hot tub: case of hot-tub lung. AB - Reports detailing Mycobacterium avium complex (MAC) infection in an immunocompetent host after exposure to hot tub (hot-tub lung) are few and have not focused on the radiographic presentation. On high-resolution computed tomography (HRCT), hot-tub lung caused by MAC infection closely resembles subacute hypersensitivity pneumonitis and is difficult to distinguish based on imaging parameters alone. Centrilobular nodules of ground-glass attenuation with beading of the interlobular septae are prominent features on HRCT. Correlation of clinical history with radiographic, pathologic, and microbiologic findings are necessary to establish a diagnosis of hot-tub lung. PMID- 12544749 TI - Mature teratoma within the posterior mediastinum. AB - Mature teratomas in the chest are typically located within the anterior mediastinum. The authors report a case of a mature teratoma in a young adult male with characteristic imaging features except for its location: the posterior mediastinum. On review of the literature, 3% to 8% are reported as confined to the posterior mediastinum and 13% have extension into other mediastinal compartments. PMID- 12544752 TI - Immediate influence of transcranial electrostimulation on pain and beta-endorphin blood levels: an active placebo-controlled study. AB - BACKGROUND: Stimulation of the antinociceptive system by noninvasive electrical current from electrodes placed on the head is a renewed method of pain relief. METHODS: We conducted a randomized, double-blind, placebo-controlled study on 20 chronic back pain patients. They were treated with either transcranial electrostimulation (TCES) or an active placebo device. Pain level and serum beta endorphin levels were measured before and after treatment. RESULTS: beta Endorphin level increased in seven of the ten patients from the treatment group and did not change in eight of ten patients from control group (P = 0.057 between groups). Pain level decreased in eight treated patients and seven control patients (significant decrease for each group, no significant difference between groups). CONCLUSIONS: Transcranial electrostimulation is a nonpharmacologic method of pain relief accompanied or mediated by beta-endorphin release. The comparable degree of the initial clinical response emphasizes the powerful placebo effect on reported pain not mediated by endorphin release. This preliminary study shows that noninvasive electrical stimulation is a safe treatment with a positive effect on beta-endorphin blood levels. PMID- 12544753 TI - Factors that influence the duration of splint wear in peripheral nerve lesions. AB - OBJECTIVE: To assess the weekly frequency and overall duration of wear for splints used to treat peripheral nerve lesions and factors that possibly influence splint wear. DESIGN: A total of 78 patients (23 women, 55 men) who had been treated with a hand splint for peripheral nerve palsy were interviewed by telephone, based on a preformulated questionnaire. The duration of wear, reasons for terminating use, the effect of the splint, the patients' assessment of splint treatment, the patients' instruction, and the patients' level of education were recorded. Survival analysis for the entire period of wear and logistic regressions to determine factors that influence splint wear were performed. Comparison of effect scores between lesions in the dominant those in the nondominant hand were carried out with a test. RESULTS: The median duration of wear was 6 mo for daytime and 4 mo for nighttime splints. Daytime splints were worn by 85% and nighttime splints by 84% of patients for 5-7 days per week. The reported effect of the splint was the only significant factor that influenced the frequency of wear. The splint was reported to be significantly more effective in the dominant hand than in the nondominant hand. Numerically, radial nerve lesions and cock-up wrist splints were most common. CONCLUSION: Regular splint wear was registered in the majority of patients and was positively influenced by a good effect of the splint, which was reported to be better in the dominant hand. PMID- 12544754 TI - Gait patterns of transtibial amputee patients walking indoors barefoot. AB - OBJECTIVE: To evaluate the gait patterns of lower limb amputee patients walking with and without shoes and to identify differences in barefoot gait patterns when using different prosthetic feet. DESIGN: Optoelectronic three-dimensional motion analysis of gait was performed on six transtibial amputees using a solid ankle cushion heel foot and a single-axis foot, both with and without shoes. RESULTS: Gait abnormalities were observed during barefoot walking when the solid ankle cushion heel foot was used. These included knee joint hyperextension of 9.9 +/- 2.0 degrees and the loss of ankle plantar flexion in the early stance phase. When the single-axis foot was used, knee flexion thrust declined from 9.9 +/- 3.7 degrees to 7.2 +/- 3.8 degrees and ankle plantar flexion decreased from 9.9 +/- 2.8 degrees to 7.0 +/- 2.1 degrees during the early stance phase. CONCLUSIONS: In transtibial amputees, significant gait abnormalities were observed during barefoot walking using the solid ankle cushion heel foot. These gait patterns improved, however, with use of a single-axis prosthetic foot, which permits a further plantar flexion after the initial contact. PMID- 12544755 TI - Contribution of accelerated body masses to able-bodied gait. AB - OBJECTIVE: The objectives of this study were to demonstrate that data from a video-based system could be used to estimate the net effect of the external forces during gait, to determine the contribution of the trunk and upper and lower limbs using their accelerated body masses, and to test the hypothesis that the thigh mainly assumed lower limb propulsion during able-bodied locomotion. METHODS: The gait of 16 able-bodied subjects was assessed using an eight-camera video-based system and two force plates. The right limb was the leading limb, and there were two trials per subject. Although data from all the body segments were used to answer the first two objectives, only right limb information was used to address the third objective. RESULTS: Pearson's coefficients of correlation and root mean square errors were calculated to determine the difference between the curves obtained from the sum of the external forces and that of the accelerated masses. These were >0.85, and the mean root mean square error was <4 N. Analyses of variance were performed on the peak forces developed by the trunk and the upper and lower limbs along each axis. Tukey's posthoc tests (P < 0.05) revealed that the trunk was the principal contributor of external forces in the frontal and transverse planes, whereas the lower limbs were found to be more important in the plane of progression. Analyses of variance and Tukey's posthoc tests (P < 0.05) were performed on the peak forces developed by each segment of the right limb. In decreasing order, the thigh, shank, and foot displayed the highest mass acceleration products in the right limb during gait. CONCLUSIONS: A video-based system was able to determine the net effect of the external forces with the summation of the mass-acceleration products during able-bodied gait. The trunk and lower limbs were the dominant body segments responsible for the production of external forces during able-bodied gait, whereas the thighs contributed more to the ground reaction force than the foot and shank for forward progression in able bodied gait. PMID- 12544757 TI - Gross motor function classification system: preliminary study for Japanese children. AB - OBJECTIVE: The purpose of the study was to produce a version of the Gross Motor Function Classification System with the same construct as the original one that would be applicable to Japanese children with cerebral palsy. DESIGN: The reliability studies were performed twice with a convenience sample (first study, 76 children with cerebral palsy; second study, 225 children with cerebral palsy). The first version of the translated Gross Motor Function Classification System was used in the first reliability study. It was revised based on the findings of the study and used in the second reliability study. Two examiners classified each child's level of gross motor function independently. We calculated kappas for individual levels and across all levels to study interrater reliability. RESULTS: The individual kappa for level V changed from 0.59 in the first reliability study to 0.87 in the second reliability study, whereas the overall kappa changed from 0.64 to 0.66. In the second reliability study, the factors that seemed to affect the reproducibility were the unfamiliar words used in the system, partial agreement of the children's status, and thinking of future levels. CONCLUSIONS: Explanation of words and a note stressing the importance of the introduction part should be added to the system for Japanese clinicians. PMID- 12544756 TI - Relationship between knee range of motion and Kellgren-Lawrence radiographic scores in knee osteoarthritis. AB - OBJECTIVE: To investigate the relationships between the joint range of motion and Kellgren-Lawrence radiographic scores of knee joint compartments in patients with knee osteoarthritis. DESIGN: Forty knees of 20 consecutive bilateral, primary knee osteoarthritis patients were examined in the study. Maximal flexion, extension, and internal and external rotation were assessed using a goniometer. Kellgren-Lawrence radiographic scores were determined for medial tibiofemoral, lateral tibiofemoral, and patellofemoral compartments of the knee joint on lateral, tangential, and standing anteroposterior radiographs. RESULTS: Statistically significant negative correlations were found between the range of motion and radiographic scores. Specifically, internal rotation was correlated with lateral compartment scores (r = -0.439, P < 0.01), and external rotation and flexion were correlated with medial compartment scores (r = -0.361, P < 0.05; r = -0.338, P < 0.05; respectively) [corrected]. Extension values were correlated with patellofemoral ( = -0.533, < 0.01), medial (r = -0.456, P < 0.01), and lateral (r = -0.327, P < 0.05) compartment scores. CONCLUSIONS: A clear relationship is present between joint range of motion and Kellgren-Lawrence radiographic scores in knee osteoarthritis patients. Examination of the joint range of motion can give information about the compartmental distribution of the disease and help the physician focus on the compartment or compartments involved when performing diagnostic and therapeutic procedures. Evaluation of the compartments of the knee joint in radiographs can be informative about the joint range of motion in knee osteoarthritis. PMID- 12544758 TI - Applicability, validity, and reliability of the Piper Fatigue Scale in postpolio patients. AB - OBJECTIVE: To identify a scale that is potentially applicable for measuring the fatigue in postpolio patients and to evaluate its validity and reliability in this population. DESIGN: Interview survey of 64 individuals with postpolio syndrome and 25 healthy controls of similar age range, with retest in a subset of postpolio patients. The sample was recruited from a postpolio support group, a postpolio clinic, and the general community. Subjects completed the Piper Fatigue Scale, the Beck Depression Inventory, and the Chalder Fatigue Questionnaire during the interview. RESULTS: Face and content validity of the Piper Fatigue Scale was established by a team of experts and by a group of postpolio patients. The postpolio subjects had significantly higher Piper Fatigue Scale scores than the healthy control subjects (P < 0.001), demonstrating extreme groups validity. Convergent validity was shown with a strong positive correlation between Piper Fatigue Scale scores and Chalder Fatigue Questionnaire scores (r = 0.80). Reliability was also demonstrated with the Piper Fatigue Scale's high internal consistency (alpha = 0.98) and strong test-retest agreement (intraclass correlation coefficient = 0.98). CONCLUSIONS: The Piper Fatigue Scale is a valid and reliable tool for measuring postpolio fatigue. This scale may be useful in other studies of postpolio fatigue, including those gauging the effectiveness of various treatments for this fatigue. PMID- 12544759 TI - Recovery of spinal cord conduction after surgical decompression for cervical spondylotic myelopathy: serial somatosensory evoked potential studies. AB - OBJECTIVE: We tested the utility of scalp-recorded median and tibial somatosensory evoked potentials (SEPs) as a measure of delineating the time course of postoperative recovery from cervical spondylotic myelopathy. DESIGN: We evaluated serial median and tibial SEP studies for 20 hands and 22 feet in 13 cervical spondylotic myelopathy patients during the first 6 mo postoperatively or longer. Serial sensory assessment of the hands and feet served to evaluate clinical correlation with the SEPs. RESULTS: The studies distinguished three recovery patterns of the SEPs based on the time course of the latencies of the N20 component for the median SEP and the P40 component for the tibial SEP. The latency started to decrease by 0.5 msec or more within 2 wk (seven hands and eight feet) or later than 2 wk (six hands and eight feet) postoperatively, or the change did not reach 0.5 msec (seven hands and six feet). In the late recovery group, the SEP improvement began as late as 10 wk postoperatively. Sensory recovery correlated with the SEP change in every group. CONCLUSIONS: An early onset of the SEP recovery predicts a favorable clinical course. The SEP may begin to improve later, and once it occurs, progressive return of function will follow. PMID- 12544760 TI - Spinal muscular atrophy type 1 quality of life. AB - OBJECTIVE: To compare healthcare professionals' assessment of the quality of life of spinal muscular atrophy type 1 children with that of the care providers for the children. DESIGN: The care providers of all 53 surviving spinal muscular atrophy type 1 children managed in one neuromuscular disease clinic were sent Likert-scale surveys of six quality of life issues and ten polar-adjective pairs. The quality of life estimations were compared with those of 67 clinicians and with those of 30 parents considering their unaffected children. RESULTS: One hundred care providers from 46 out of the 53 families (87%) responded. Although the clinicians' mean estimate of the children's quality of life was 2.85 +/- 0.2/10, the care providers' estimate was 7.81 +/- 0.2/10 (P < 0.0001). The care providers also found life with the children to be satisfying (6.0 +/- 0.2/7), interesting (6.6 +/- 0.1/7), friendly (6.1 +/- 0.1/7), enjoyable (6.3 +/- 0.1/7), worthwhile (6.7 +/- 0.1/7), full (6.6 +/- 0.1/7), hopeful (5.9 +/- 0.2/7), and rewarding (6.4 +/- 0.1/7), and they estimated the children to be happy (8.5 +/- 0.2/10) and their lives worth living (9.6 +/- 0.1/10). However, 69 of 104 felt that their lives were hard rather than easy, and 56 of 104 reported feeling tied down rather than free. Although the effort they felt for raising the child was high (8.3 +/- 0.3 by comparison with 5 for an unaffected child), the burden they felt in doing so was not (5.8 +/- 0.3/5). When asked whether they would or would not recommend ventilator use, 31 clinicians (45.5%) indicated they would, 24 (36.4%) would not, and 12 (18.2%) chose not to respond to this question. Care provider responses did not differ significantly from the responses of the parents of unaffected children except for the easy/hard semantic differential (care providers, 3.80 +/- 1.75 controls, 5.27 +/- 1.14, < 0.001). CONCLUSIONS: Although there is a widespread perception that spinal muscular atrophy type 1 children have a poor quality of life, this perception is not shared by their care providers. PMID- 12544761 TI - Functional recovery and length of stay after hip fracture in patients with neurologic impairment. AB - OBJECTIVE: To evaluate the functional recovery and the length of stay after hip fracture in patients with neurologic impairment. DESIGN: A total of 577 inpatients with hip fracture consecutively admitted to our rehabilitation hospital were included in this retrospective study. A total of 71 of 577 were affected by neurologic impairment caused by stroke with hemiplegia (n = 37), Parkinson's disease (n = 25), or other diseases (n = 9). RESULTS: Mean Barthel index was significantly lower in the patients with neurologic impairment than in the controls: 10.8 (95% confidence interval, 5.9-15.6; P < 0.001) at admission and 13.1 (95% confidence interval, 5.55-20.65; P < 0.001) at discharge. Multiple regression including eight confounding variables showed that neurologic impairment was negatively associated with the Barthel index. However, the mean increase in Barthel index through the course of rehabilitation was not affected by neurologic impairment. The length of stay was significantly higher in the patients with neurologic impairment, 3.84 days (95% confidence interval, 0.51 7.17; P < 0.05), and multiple regression showed that neurologic impairment was positively associated with the length of stay. CONCLUSIONS: After hip fracture, the presence of neurologic impairment was associated with lower Barthel index and longer length of stay, but it did not affect the increase in Barthel index due to a course of rehabilitation. PMID- 12544762 TI - The ethics of advance directives: a rehabilitation perspective. AB - Advance directives, including living wills and durable healthcare powers of attorney, have achieved broad acceptance by the healthcare system in the United States. Living wills may include provisions for limitation of care in the event of severe disability. These provisions pose ethical concerns in view of societal misconceptions of the quality of life of individuals with disabilities and the inability of people to predict their own capacity to adapt successfully to a disability. Greater reliance on durable healthcare powers of attorney for situations involving disability is proposed, with an emphasis on improving the education of healthcare proxies designated through this mechanism in the quality of life experienced by people with disabilities. PMID- 12544763 TI - Rehabilitation treatment options for a patient with paraneoplastic cerebellar degeneration. AB - Paraneoplastic cerebellar degeneration is a neurologic syndrome associated with carcinoma. Medical management is limited to treatment of the underlying malignancy with excision, plasmapharesis, immunosuppression, or chemotherapy. We report on the case of a woman with paraneoplastic cerebellar degeneration who showed improvements in functional mobility after 3 wk of comprehensive inpatient rehabilitation. This case demonstrates the functional improvements of one patient with paraneoplastic cerebellar degeneration after intensive rehabilitation and describes the specific treatment interventions utilized. PMID- 12544765 TI - Screening for neuroblastoma: the final chapters. PMID- 12544766 TI - Neurocognitive functioning after cancer: is that all there is to QOL? PMID- 12544767 TI - High-risk neuroblastoma in Ontario: a report of experience from 1989 to 1995. AB - PURPOSE: We did a population-based study of children with high-risk neuroblastoma to determine their survival and look for factors that had an impact on survival. METHODS: We carried out a retrospective cohort study of patients diagnosed in Ontario from 1989 to 1995. 162 cases of neuroblastoma were diagnosed in the province with 70 (43%) considered high-risk: all were older than one year of age, with 15 patients classified as International Neuroblastoma Staging System (INSS) stage 3, and 55 INSS stage 4. RESULTS: Stage 3 patients did significantly better than Stage 4 patients with a 5-year overall survival of 67.7% and 22.7% respectively (P = 0.015). In stage 4 patients achieving at least a partial response to up-front therapy and surviving for at least 9.5 months after diagnosis (the median time to transplant), there was no difference in survival between the 19 transplant patients and the 17 treated with chemotherapy alone (P = 0.75). However, patients transplanted by peripheral stem cell (PSC) collection did significantly better than both the bone marrow transplantation (P = 0.002) and chemotherapy-alone group (P = 0.047). There was a significant difference in up-front chemotherapy and use of radiation in the three groups (P < 0.001 and P = 0.01 respectively), but no difference in the incidence of bone and bone marrow metastases, MYCN amplification or unfavorable histology. CONCLUSIONS: In this nonrandomized study, we found that stage 4 neuroblastoma patients alive more than 9.5 months after diagnosis, with at least a partial response to initial therapy, did significantly better with PSC transplant compared with bone marrow transplantation or chemotherapy alone. PMID- 12544768 TI - False-positive results in neuroblastoma screening: the parents' view. AB - PURPOSE: One of the major problems of any screening program is the occurrence of false-positive results. For neuroblastoma screening, little information is available on the psychological consequences for parents whose children had false positive results that made further clinical evaluation necessary. It was the aim of this study to evaluate the parents' view by a semistructured interview. PATIENTS AND METHODS: Among 267,302 infants screened in the Austrian study between 1991 and 1999, 19 had to be considered as repeatedly false-positive (no clinical evidence of neuroblastoma). Sixteen of 19 parent pairs could be reached by phone and were interviewed separately by use of a semistructured questionnaire to evaluate for psychological consequences resulting from the screening result. RESULTS: The psychological burden appeared to be small during the initial screening procedure, but it increased significantly through hospital admission and was then described as severe by 19 of the 32 parents. CONCLUSIONS: Investigators should be aware of the psychological consequences of hospital admission for tumor screening in children. In ongoing neuroblastoma screening studies, laboratory methods as well as cutoff limits should be selected carefully. PMID- 12544769 TI - Effects of amifostine on clonogenic mesenchymal progenitors and hematopoietic progenitors exposed to radiation. AB - PURPOSE: To determine the radiation sensitivities of mesenchymal progenitors and hematopoietic progenitors, and to determine the in vitro effects of amifostine on hematopoietic and mesenchymal progenitors exposed to radiation. METHODS: Radiosensitivity of mesenchymal progenitor cells was determined by exposing marrow low-density cells to radiation at doses of 100 to 800 cGy. Mesenchymal cell colonies were established by plating 2.5 x 10(5) marrow low-density cells in long-term marrow culture medium (LTCM). The size, frequency, and cellular composition of the mesenchymal progenitor cells were scored after 14 days of incubation. Mesenchymal progenitor cells were subdivided into progenitors forming fibroblast and adipocyte mixed colonies (CFU-FA), and pure fibroblast colonies (CFU-F). Hematopoietic progenitors were assessed by methylcellulose-based assay. RESULTS: Radiation at 100 cGy caused a mild decrease in CFU-F and CFU-FA derived colonies by 12% and 13%, respectively; 200 cGy decreased CFU-F by 36% and CFU-FA by 52%; 400 cGy decreased CFU-F by 50% and CFU-FA by 86%; and 600 cGy decreased CFU-F by 24%, with total absence of CFU-FA. Pretreatment with amifostine protected 100% of CFU-F at 100 and 200 cGy, 84% at 400 cGy, 46% at 600 cGy, and 14% at 800 cGy. With CFU-FA colonies amifostine pretreatment provided only minimal radioprotection. For hematopoietic progenitors radiation at 100 cGy reduced CFU-GM by 74% but had no significant effect on CFU-GEMM and BFU-E. Radiation at 200 cGy decreased CFU-GEMM by 72%, BFU-E by 54%, and CFU-GM by 84%; 400 cGy further decreased CFU-GEMM by 83%, BFU-E by 81%, and CFU-GM by 93%. Pretreatment with amifostine resulted in twofold stimulation of CFU-GEMM and BFU E colonies. All BFU-E colonies were protected up to 200 cGy. For CFU-GEMM amifostine pretreatment resulting in 68% at 200 cGy and 31% at 400 cGy. For CFU GM colonies it was 54% at 100 cGy, 32% at 200 cGy, and 12% at 400 cGy. CONCLUSIONS: Mesenchymal progenitor cell subpopulations are differentially sensitive to radiation. Amifostine protects both mesenchymal and hematopoietic progenitors against radiation injury, though the level of protection appears to be dependent upon the sensitivities of these progenitor cells to radiation. Amifostine is a potent stimulant of BFU-E and CFU-GEMM progenitor colonies. PMID- 12544770 TI - Her-2/neu expression in osteosarcoma increases risk of lung metastasis and can be associated with gene amplification. AB - The purpose of this study was to investigate whether Her-2/neu expression at diagnosis of osteosarcoma could provide biologic and prognostic information that predicts the risk of pulmonary metastases and outcome. Human epidermal growth factor (Her-2/neu) expression in 25 initial pretreatment osteosarcoma biopsies and 12 posttreatment pulmonary metastatic osteosarcoma resection specimens was assessed by standard immunohistochemical techniques on formalin-fixed paraffin embedded tissue. As a screening analysis to determine if gene amplification may be a mechanism for increased Her-2/neu expression, FISH analysis was conducted on seven Her-2/neu immunostain-positive samples and five Her-2/neu immunostain negative samples. Cytoplasmic Her-2/neu reactivity was identified in 11/25 (44%) of primary tumors and in 7/12 (58%) resection specimens from pulmonary metastases. Cytoplasmic Her-2/neu expression was associated with shorter overall metastasis-free survival. Her-2/neu gene amplification was identified by FISH analysis in six of the seven immunostain-positive samples but was also identified in two of the five immunostain-negative samples. Her-2/neu expression in patients with osteosarcoma is associated with an increased risk of metastasis and may define a subset of patients with a more aggressive tumor phenotype. Her-2/neu gene amplification may provide a mechanism for Her-2/neu overexpression in certain cases of osteosarcoma. Whether Her-2/neu expression influences outcome needs to be examined further in a prospective fashion. The hope is that Her-2/neu expression will identify patients who may benefit from the addition of directed biologic therapy. PMID- 12544771 TI - Whole blood international normalization ratio measurements in children using near patient monitors. AB - PURPOSE: To report a comparison of international normalization ratio (INR) measurements on four near-patient (point-of-care or bedside) whole blood INR monitors in children. PATIENTS AND METHODS: The INR results from 19 ambulatory pediatric subjects (30 hospital visits) receiving warfarin sodium were analyzed on four near-patient monitors and compared with plasma INR measurements on the laboratory CA-1000 Analyze. The instruments evaluated were CoaguChek, Hemochron Jr. Signature, ProTime Microcoagulation System, and RapidpointCoag. RESULTS: The INR measurements ranged from 1.05 to 5.25. Over the entire INR range, the near patient instrument with the least bias relative to the CA-1000 was the RapidpointCoag (r(2) = 0.923). The correlations (r(2)) of the CoaguCheck, Hemochron Jr., and ProTime were 0.877, 0.834, and 0.885, respectively. Precision studies involved repeated analysis of one nonmedicated adult (mean CA-1000 INR = 0.908) and one adult receiving oral anticoagulation therapy (mean CA-1000 INR = 2.42). The coefficient of variation on the near-patient monitors for both adult volunteers ranged from 4.9% to 22.3%. Bilirubin levels up to 20 mg/dL did not interfere in any of the methods. CONCLUSIONS: Near-patient testing whole blood INR monitors offer acceptably accurate and precise measurements. Values obtained on near-patient monitors may vary considerably from the reference method, and data obtained should serve as a supplement to, but not a replacement for, routine clinical laboratory measurements. PMID- 12544772 TI - Recombinant tissue plasminogen activator (alteplase) for restoration of function to occluded central venous catheters in pediatric patients. AB - PURPOSE: To evaluate the safety and efficacy of alteplase for restoring function to occluded central venous catheters in a pediatric population. PATIENTS AND METHODS: A phase III, open-label, single-arm, multicenter trial was performed in 995 adult and pediatric patients with dysfunctional nondialysis catheters and ports. This report is a subset analysis of subjects between 2 and 18 years of age (N = 122) who were enrolled in the study. Alteplase (2 mg/2 mL) was instilled into the dysfunctional catheter lumen and assessed at 30 and 120 minutes. Subjects weighing > or =30 kg received 2 mL of alteplase; subjects <30 kg received 110% of the internal lumen volume (not exceeding 2 mL). Alteplase dosing was repeated once after 120 minutes if the catheter remained dysfunctional. The primary safety endpoint was the rate of intracranial hemorrhage (ICH) within 5 days of treatment. RESULTS: The overall efficacy following up to two instilled doses of alteplase was 87%. In 70 patients (57%), restoration of catheter flow occurred by 30 minutes following a single dose of alteplase. Restoration of function was related to the duration of occlusion (P = 0.04). For catheters with occlusions of 0, 1 to 14, and >14 days duration, the efficacy was 91%, 78%, and 60%, respectively. Success was independent of the patient's age, sex, body weight, CVC type, or catheter age. There were no cases of death, ICH, major bleeding episodes, or embolic events attributable to treatment. CONCLUSIONS: An alteplase regimen of up to two 2-mg doses is safe and effective for restoration of function to occluded central venous catheters in a pediatric population. PMID- 12544773 TI - Chlamydia pneumoniae and acute chest syndrome in patients with sickle cell disease. AB - PURPOSE Few studies address the association of Chlamydia pneumoniae infection with pulmonary disease and outcome in patients with underlying pathology such as sickle cell disease (SCD). SCD patients are susceptible to the pulmonary disorder known as acute chest syndrome (ACS), where the etiology remains ill defined. The purpose of this study was to analyze the clinical course and outcome of C. pneumoniae-associated ACS among SCD patients as part of the National Acute Chest Syndrome Study. PATIENTS AND METHODS: This was a longitudinal study of SCD patients presenting with ACS to multiple U.S. medical centers. Two hundred ninety six SCD patients who developed ACS were tested by PCR for C. pneumoniae and by standard techniques for other respiratory pathogens. These infections were evaluated for association with ACS, clinical course, and complications. RESULTS: Forty-one (14%) patients with first episodes of ACS were PCR positive for C. pneumoniae. Compared with other infections, C. pneumoniae-infected patients were older, were more likely to present with chest pain, and had higher hemoglobin levels at diagnosis. Both groups had similar rates of respiratory failure and prolonged hospitalization. Of the 89 patients with single-pathogen infections, 27 (30%) were due to C. pneumoniae, 21% to Mycoplasma pneumoniae, 10% to RSV, 4% to Staphylococcus aureus, and 3% to Streptococcus pneumoniae. CONCLUSIONS: C. pneumoniae was the most prevalent pathogen in this study of ACS and was responsible for significant morbidity. Additional research is required to develop effective treatment guidelines for ACS. PMID- 12544774 TI - Development of disease-specific health-related quality-of-life instruments for children with immune thrombocytopenic purpura and their parents. AB - PURPOSE: Immune thrombopenic purpura (ITP) is an important childhood hematologic disorder that is often frightening to patients and their parents because of its acute onset and bleeding symptoms. There is no consensus on the management of ITP in children. Pediatric hematologists have differing management philosophies, yet most, explicitly or implicitly, incorporate into their management approach the potential impact on the child's and family's quality of life. There is no validated ITP-specific health-related quality-of-life instrument for use with children with ITP, nor is there one to evaluate the burden experienced by their parents. ITP is usually a self-limited disorder. With current controversy over management approaches, an evaluation of the disease burden experienced by the child and the family may assist with the assessment of alternative treatment approaches. METHODS: Using standard clinimetric methodology, 88 children with acute or chronic ITP, along with their parents, participated in the development of the instruments. RESULTS: The 26-item ITP-Child Quality-of-Life Questionnaire includes five domains: treatment side effect-related, intervention-related, disease-related, activity-related, and family-related. This instrument can be used as a self-completed instrument for most children older than 7 years or as a proxy-completed instrument by parents of children younger than 7 years. The 26 item ITP-Parental Burden Quality-of-Life Questionnaire includes six domains: concerns related to diagnosis/investigation, treatment/disease monitoring, monitoring of child's activities, interference with daily life, disease outcome, and emotional impacts. CONCLUSIONS: The first steps of the development of these formally developed instruments are complete. The instruments are available for study to validate and test their responsiveness through use in clinical research studies. Such instruments are increasingly recognized as important for comprehensive measurement of patient outcomes in this and other areas of pediatric hematology/oncology practice. PMID- 12544775 TI - Bone marrow hypoplasia during Brucella infection. AB - Pancytopenia, although mainly reported in adults, has also been described in children with brucellosis. However, bone marrow hypoplasia is a rare feature of the infection. An 11-year-old boy was admitted with fever, vomiting, and abdominal pain of 10 days' duration. On physical examination, pallor and high fever were detected in the absence of lymphadenopathy and hepatosplenomegaly. His hemoglobin was 8.6 g/dL, white blood cell count 1,100/mm(3), neutrophil count 500/mm(3), platelets 56,000/mm(3), and reticulocytes 0.1%. Hypocellular bone marrow was found by aspiration, and bone marrow biopsy revealed hypocellularity. The agglutination titer was greater than 1/640. Trimethoprim/sulfamethoxazole was prescribed. His fever subsided and pancytopenia subsequently improved. Pancytopenia associated with brucellosis is attributed to hypersplenism, hemophagocytosis, and granulomatous lesions of the bone marrow, which is usually hypercellular. Bone marrow hypoplasia is rarely reported and should be kept in mind in the etiology of aplastic anemia in a country where brucellosis is frequently encountered. PMID- 12544776 TI - Extramedullary relapse in the left proximal femur with Philadelphia chromosome positive acute lymphoblastic leukemia after allogeneic bone marrow transplantation. AB - We describe a rare presentation of extra-medullary relapse in an adolescent boy with Philadelphia chromosome-positive acute lymphoblastic leukemia who had undergone allogeneic bone marrow transplantation after first remission. In spite of enduring bone marrow remission, the patient experienced a local relapse in the left proximal femur within 3 years of the transplant. The findings from radiography, bone scintigraphy, and chimerism analysis with short tandem repeats as well as bone marrow aspirates taken via the iliac crests were indeterminate. Magnetic resonance imaging at the onset of hip pain was characterized by decreased signal intensity of the left proximal femur, a finding characteristic of bone marrow edema. Confirmation of extra-medullary relapse of the proximal femur was delayed until histologic proof of the computed tomography-guided biopsy samples was obtained. Overt bone marrow relapse was identified 14 months later. Reestablishment of normal donor hematopoiesis was achieved with reinduction chemotherapy. PMID- 12544777 TI - Acute blindness as a presenting sign in childhood non-Hodgkin lymphoma. AB - Non-Hodgkin lymphoma is the third most common malignancy in childhood. Occasionally, a patient with lymphoma may present with neuroophthalmologic symptoms and signs. It is usually associated with 3rd, 4th, and 6th cranial nerve palsies. Here, we report three children presenting with loss of vision. Two of them were cured with modified LMB-89 (lymphoma malignant B) chemotherapy regimen and radiotherapy. In one patient, loss of vision returned to normal in the left eye but persisted in the right eye. The second patient suffered from permanent visual loss, although the primary disease responded well to treatment. Unfortunately, the third patient died at an early phase of treatment. Acute blindness as a presenting symptom is rare in lymphoma. Early diagnosis and treatment is critical to increase the survival rate and to improve the quality of the vision. PMID- 12544778 TI - A chemosensitive pediatric extraosseous osteosarcoma: case report and review of the literature. AB - Osteosarcoma arising in soft tissues is exceedingly rare in children. The tumor most often affects older adults, involves the lower extremity, responds poorly to chemotherapy, and carries a grave prognosis. The authors describe a 12-year-old girl with an extraosseous osteosarcoma of the left sternocleidomastoid muscle with pulmonary metastases. The patient responded well to neoadjuvant chemotherapy and remains disease-free nearly 3 years after her initial diagnosis. The authors review available information about this disease in children and adults. Children with extraosseous osteosarcoma may have a more favorable response to treatment than adults; thus, a curative approach using combined modality therapy appears warranted. PMID- 12544779 TI - High-dose chemotherapy and autologous peripheral blood stem cell rescue in a patient with pleuropulmonary blastoma. AB - Pleuropulmonary blastoma (PPB) is a rare and aggressive malignant tumor of the lung. Approximately 80 cases of PPB have been published, and in only three cases high-dose chemotherapy with autologous hematopoietic stem cell transplantation (HSCT) was applied. A 5-year-old girl presenting with cough, fever, and shortness of breath was referred to the authors in March 1999. A computed tomography scan of the chest showed a tumor mass in the left hemithorax. The lesion was biopsied and the histopathologic report suggested the diagnosis of PPB. The patient received chemotherapy comprising vincristine, actinomycin D, and cyclophosphamide with only a minor response, and treatment was switched to ifosfamide, carboplatin, and etoposide, which produced a partial response. Tumor resection was performed, but margins were positive for PPB. Due to the high risk of recurrence, the authors elected to administrate high-dose chemotherapy using melphalan, etoposide, and carboplatin, followed by autologous HSCT. The patient achieved complete hematologic recovery, and reimaging after HSCT showed no evidence of disease. She relapsed 4 months later and died about 9 months after the completion of high-dose therapy. The role of high-dose chemotherapy and autologous HSCT is likely to be limited in PPB. PMID- 12544780 TI - t(5;9)(q11;q34): a novel familial translocation involving Abelson oncogene and association with hypereosinophilia. AB - A 6-year-old girl with hypereosinophilia was found to have a familial constitutional translocation t(5;9)(q11;q34). Flow cytometry and gene rearrangement studies did not show any clonal T-helper cell proliferation. Presence of cryptic Philadelphia translocation was ruled out by reverse transcription polymerase chain reaction. Abelson oncogene translocation on chromosome 5 was confirmed by fluorescent in situ hybridization. This is the first example of a familial translocation involving the abelson oncogene and association with hypereosinophilia. The authors discuss a novel mechanism of hypereosinophilia involving the hybrid product of the abelson oncogene with an unknown partner gene on chromosome 5 (probably granzyme-A). PMID- 12544781 TI - Polyclonal proliferation of lymphocytes containing the epstein-barr virus genome in a patient dying of myocarditis in chronic active Epstein-Barr virus infection. AB - An 11-year-old boy had intermittent fever and hepatosplenomegaly. The diagnosis of chronic active Epstein-Barr virus (EBV) infection was established from an abnormal pattern of anti-EBV antibody; EBV was detected in bone marrow cells. Immunochemotherapy alleviated the patient's symptoms. However, when a subsequent oral prednisolone dose was tapered, heart failure ensued and he died. Autopsy findings demonstrated that myocarditis resulted from infiltrating T lymphocytes with the EBV genome and a benign histologic appearance. A clonality study of T lymphocytes indicated no such evidence of monoclonality. EBV-infected T cells play an important role in the pathogenesis of myocarditis in chronic active EBV infection. PMID- 12544782 TI - Pulmonary cytolytic thrombi: unusual complication of hematopoietic stem cell transplantation. AB - Pulmonary complications of hematopoietic stem cell transplantation (HSCT), including peripheral blood stem cell transplantation (PBSCT) and bone marrow transplantation, are frequent and often life-threatening. Differentiating acute infectious from noninfectious pulmonary complications is difficult but critical for proper treatment. The authors describe an 11-year-old boy who developed a sudden fever and cough associated with a normal chest radiograph 2 months after successful haploidentical PBSCT for severe aplastic anemia. High-resolution chest computed tomography revealed numerous tiny peripheral pulmonary nodules. Lung biopsy demonstrated an unusual occlusive thrombotic vascular lesion associated with hemorrhagic infarction without evidence of infection. The thrombi were composed of intensely basophilic granular material recently described as "cytolytic" thrombi. Symptoms and chest computed tomography improved rapidly following intravenous corticosteroids and cyclosporin. However, the patient subsequently died of rapidly progressive pulmonary hypertension. Our patient illustrates the importance of considering this noninfectious complication in the acute pulmonary disorders associated with HSCT as this condition may represent a pulmonary manifestation of acute graft-versus-host disease. PMID- 12544783 TI - Subcutaneous interleukin-2 therapy in a child with vertically transmitted HIV-1 infection. PMID- 12544784 TI - A guest editorial: new insights to shoulder dystocia and brachial plexus palsy. PMID- 12544785 TI - Molecular epidemiology of preeclampsia. AB - Numerous articles have been published that address the possible genetic influences on the development of preeclampsia (PE). However, to our knowledge, a complete review of the results has not yet been completed. We undertook a MEDLINE search to identify English-language articles published after January 1, 1990 that examined the possible role of specific genes in the etiology of PE. After a brief introduction and a concise review of the prevailing etiologic hypotheses, we have categorized the candidate genes into six categories, based on their hypothesized role in PE etiology. The purpose of this paper is to review the literature, comment on its quality, and provide a reference for researchers interested in the molecular epidemiology of preeclampsia. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader will be able to list the prevailing etiologic hypotheses of preeclampsia, to outline the published data on possible genetic influence on the development of preeclampsia, and to clearly state the definition of preeclampsia. PMID- 12544786 TI - Long-term physical and psychological health consequences of induced abortion: review of the evidence. AB - Induced abortion is a prevalent response to an unintended pregnancy. The long term health consequences are poorly investigated and conclusions must be drawn from observational studies. Using strict inclusion criteria (study population >100 subjects, follow up >60 days) we reviewed an array of conditions in women's health. Induced abortion was not associated with changes in the prevalence of subsequent subfertility, spontaneous abortion, or ectopic pregnancy. Previous abortion was a risk factor for placenta previa. Moreover, induced abortion increased the risks for both a subsequent preterm delivery and mood disorders substantial enough to provoke attempts of self-harm. Preterm delivery and depression are important conditions in women's health and avoidance of induced abortion has potential as a strategy to reduce their prevalence. Only review articles including the single published meta-analysis exploring linkages between abortion and breast cancer were relied upon to draw conclusions. Reviewers were mixed on whether subsequent breast neoplasia can be linked to induced abortion, although the sole meta-analysis found a summary odds ratio of 1.2. Whatever the effect of induced abortion on breast cancer risk, a young woman with an unintended pregnancy clearly sacrifices the protective effect of a term delivery should she decide to abort and delay childbearing. That increase in risk can be quantified using the Gail Model. Thus, we conclude that informed consent before induced abortion should include information about the subsequent risk of preterm delivery and depression. Although it remains uncertain whether elective abortion increases subsequent breast cancer, it is clear that a decision to abort and delay pregnancy culminates in a loss of protection with the net effect being an increased risk. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader will be able to define the terms and, to outline the epidemiologic problems in studying the long-term consequences of abortion, and to list the associated long term consequences of abortion. PMID- 12544787 TI - Cautery of the orbital septum during blepharoplasty. AB - PURPOSE: Application of a grid of electrocautery to the orbital septum to treat anteriorly prolapsed eyelid fat pads during blepharoplasty has been previously described. A review of patients who underwent this technique was performed to determine if this technique led to a higher incidence of postoperative eyelid retraction. METHODS: A retrospective case series of patients who underwent application of a grid of electrocautery to the orbital septum during blepharoplasty from 1979 to 1999 was reviewed. RESULTS: There were 1492 patients (3018 eyelids) who underwent an application of a grid of electrocautery to the orbital septum during this period. Five hundred twenty-nine patients (1036 eyelids) underwent upper blepharoplasty, 91 patients (174 eyelids) underwent lower blepharoplasty, 149 patients (596 eyelids) underwent combined upper and lower blepharoplasty, and 723 patients (1212 eyelids) underwent a combined upper blepharoplasty and ptosis repair. All patients were followed for at least 3 months. Follow-up ranged from 3 months to 20 years. No patient had postoperative eyelid retraction. CONCLUSIONS: The application of a grid of electrocautery to the orbital septum during blepharoplasty to treat anteriorly prolapsed eyelid fat pads is effective, safe, and does not lead to late postoperative eyelid retraction. PMID- 12544788 TI - Anchored flaps in post-Mohs reconstruction of the lower eyelid, cheek, and lateral canthus: avoiding eyelid distortion. AB - PURPOSE: To describe a system of post-Mohs reconstruction that addresses lower eyelid susceptibility to unopposed tractional, cicatricial, and gravitational forces. Large flaps are anchored to fixed tissue to avoid transmitting flap tension to the eyelids. METHODS: This is a retrospective, cohort study drawn from approximately 40 patients with post-Mohs defects of the nonmarginal lower eyelid, cheek, and lateral canthus. Surgical intervention involved horizontally oriented, relaxed skin tension line-designed advancement flaps, usually with eyelid margin stabilization. The use of anchoring sutures and any requirement for flap-graft combinations were based on defect size and the elasticity of adjacent tissues. RESULTS: Anchoring eyelid and cheek flaps to underlying periosteum permitted broad flap dissection and advancement without distortion, as the semimobile eyelid and canthi were protected from the resulting flap tension. Anticipated defect size limits for flap reconstruction often were exceeded. Defects too broad for flap reconstruction alone could be downsized, leaving a relatively small area for graft resurfacing. CONCLUSIONS: Anchored cheek flaps extend recognition of the continuity of the lower eyelid and midface to the primary reconstruction of tumor-free defects, and they address the relation by restoring deep attachments that minimize eyelid and canthal dystopia. PMID- 12544789 TI - Graft contraction: a comparison of acellular dermis versus hard palate mucosa in lower eyelid surgery. AB - PURPOSE: To compare graft contraction rates of acellular dermis versus hard palate mucosa when used as free spacer grafts in lower eyelid surgery and to provide clinical outcome data. METHODS: A prospective, nonrandomized clinical trial involving the placement of 19 spacer grafts in the lower eyelids of 14 patients was performed. Indications for spacer graft placement included lower eyelid retraction and mildly contracted socket. Patients with lower eyelid retraction also underwent an endoscopic subperiosteal midface lift. For all procedures, the height of each graft was measured during and after surgery. The amount of contraction was measured for each graft, and a mean was calculated for each spacer material. The clinical success was evaluated for all procedures, based on improvement of the functional concern being addressed. RESULTS: The mean graft contraction rate was 57% for the acellular dermis and 16% for the hard palate mucosal grafts (P <0.005). Of the 7 procedures using acellular dermis for lower eyelid retraction, 6 were considered a success, and 1 was considered a partial success. Of the 6 procedures using hard palate for lower eyelid retraction, 5 were considered a success, and 1 was considered a failure unrelated to the graft. Of the 5 procedures with acellular dermis used for mildly contracted socket, 2 were considered a success, 2 were considered a partial success, and 1 was considered a failure because of graft contraction. The one case using hard palate for mildly contracted socket was considered a success. CONCLUSIONS: Acellular dermis contracts significantly more than hard palate mucosa when used as a lower eyelid spacer graft. Acellular dermis and hard palate mucosa were both associated with a high rate of clinical success in all categories except for patients with a mildly contracted socket who received acellular dermis; more than 60% of these patients (n=5) had only partial success or failure caused by graft contraction. PMID- 12544790 TI - Medial rectus muscle injuries associated with functional endoscopic sinus surgery: characterization and management. AB - OBJECTIVE: To characterize and evaluate treatment options for medial rectus muscle (MR) injury associated with functional endoscopic sinus surgery (FESS). DESIGN: Retrospective interventional case series. PARTICIPANTS: A total of 30 cases were gathered from 10 centers. METHODS: Cases of orbital MR injury associated with FESS surgery were solicited from members of the American Society of Ophthalmic Plastic and Reconstructive Surgery (ASOPRS) through an e-mail discussion group. MAIN OUTCOME MEASURES: Variables assessed included patient demographics, computerized tomography and operative findings, extent of MR injury and entrapment, secondary orbital/ocular injuries, initial and final ocular alignment and ductions, and interventions. RESULTS: A spectrum of MR injury ranging from simple contusion to complete MR transection, with and without entrapment, was observed. Four general patterns of presentation and corresponding injury were categorized. CONCLUSIONS: Medial rectus muscle injury as a complication of FESS can vary markedly. Proper characterization and treatment are important, particularly with reference to the degree of direct MR injury (muscle tissue loss) and entrapment. Patients with severe MR disruption can benefit from intervention but continue to show persistent limitation of ocular motility and functional impairment. Prevention and early recognition and treatment of these injuries are emphasized. PMID- 12544791 TI - Treatment of prominent eyes with orbital rim onlay implants: four-year experience. AB - PURPOSE: Different approaches have been proposed to address the aesthetic or reconstructive challenge associated with relatively prominent eyes. Operations that address the soft tissues alone are prone to failure if the underlying orbital bony relationships are not addressed. Orbital rim advancement can serve as a supplement to orbital decompression in this setting or as an alternative for patients who may not maximally benefit from decompression surgery. METHODS: We report our 4-year experience with porous polyethylene orbital rim onlay grafts used to address relative proptosis in 24 patients. RESULTS: All patients had some degree of subjective and objective improvement. Proptosis decreased an average of 4.65 mm, with a range of 3 to 9 mm, based on single-observer Hertel exophthalmometry measurements (5.2 mm in those with concomitant decompression). Lagophthalmos was also improved in all patients with preoperative inadequacy in eyelid closure. Interpalpebral fissure size was reduced 1.3 mm on average, with a range of 0.5 to 6 mm (2 mm in those with concomitant decompression). Average follow-up was 41 months, with a range of 7 to 70 months. In 3 cases, we noted postoperative lower eyelid retraction with eyelid adhesion to the implant; possible risk factors for this complication included reoperative cases and simultaneous eyelid reconstruction with hard palate grafts. CONCLUSIONS: The porous polyethylene orbital rim onlay implant offers a relatively simple and effective surgical technique for the treatment of symptomatic relative proptosis. It can be used alone or in combination with other techniques including midface lift, lower eyelid retractor recession, and orbital decompression. To be effective, the implant should be placed so that it is flush with or overlapping the orbital rim; lateral displacement negates the effect of the implant in improving the eyelid/globe relationship. Postoperative eyelid retraction with tethering to the implant is a potential risk of the onlay implant, and although it may not be possible to avoid this in all cases, surgeries should be designed to minimize postoperative eyelid retraction. PMID- 12544793 TI - The use of vicryl mesh in 200 porous orbital implants: a technique with few exposures. AB - PURPOSE: To report the results of a wrapping technique for porous orbital implants by using polyglactin 910 (Vicryl) mesh (Ethicon Inc., Somerville, NJ, U.S.A.). METHODS: We retrospectively reviewed the records of 200 consecutive patients from one author's practice who received a polyglactin 910 mesh-wrapped porous orbital implant after enucleation or as a secondary implant between October 1, 1996, and April 15, 2001. We recorded potential problems that might be attributed to polyglactin 910 mesh both before pegging (excessive inflammatory response to the material, conjunctival thinning, and implant exposure) and after pegging (exposure of the implant around the sleeve, conjunctival thinning, and implant exposure other than adjacent to the peg). RESULTS: One hundred twenty-two men and 78 women underwent placement of a polyglactin 910 mesh-wrapped porous orbital implant. The average age at the time of implantation was 48.9 years (range, 11 to 85 years). The average follow-up interval in the 200 patients was 19.4 months (range, 2 to 80 months). Thirteen of the 200 patients had less than 6 months of follow-up, leaving 187 patients with an average follow-up of 20.5 months (range, 6 to 80 months). There were 76 primary enucleations and 124 secondary orbital implants. Thirty-seven patients received a Bio-Eye hydroxyapatite implant (HA) (Integrated Orbital Implants, San Diego, Calif), 97 received a synthetic FCI hydroxyapatite implant (FCI, Issy-Les-Moulineaux, France), and 66 received a Bioceramic implant (aluminum oxide-Al2O3) (FCI, Issy Les-Moulineaux, France). One hundred fourteen patients (57%) underwent peg placement. The average time to pegging was 9.9 months (range, 6 to 16 months). Before pegging, 4 of 187 patients (2.1%) had implant exposure. Three of these exposures followed secondary orbital implant placement (2 Bio-Eye HA, 1 synthetic FCI3 HA) and one followed an enucleation (synthetic FCI3 HA). Two patients required a temporalis fascia graft and one required a scleral patch; the remaining defect closed spontaneously. One patient had conjunctival thinning 6 months after orbital implantation, which remained stable with no frank exposure for 36 months. No patient had excess socket inflammation. After peg placement, 3 additional patients had exposure of the implant around the peg site. There were no cases of conjunctival thinning or exposure of the implant other than adjacent to the peg site. CONCLUSIONS: Polyglactin 910 mesh is an excellent option as a wrapping material for porous orbital implants. It is simple to use, readily available, eliminates the need for donor tissue, does not require a second operative site, and it is less expensive than other currently available wrapping materials. We attribute our high success rate to our technique, which emphasizes proper placement of the implant within the Tenon space, suturing the extraocular muscles anterior to their normal anatomic sites, and meticulous closure of the Tenon capsule and conjunctiva in separate layers. PMID- 12544792 TI - Hydroxyapatite orbital implant vascularization assessed by magnetic resonance imaging. AB - PURPOSE: To report hydroxyapatite (HA) implant enhancement patterns on magnetic resonance (MR) images at varying time intervals after implantation. METHODS: We retrospectively reviewed the records of 45 consecutive patients(from one author's practice) who underwent an MR imaging study 2 to 157 months after HA orbital implant placement. Implant fibrovascular ingrowth was assessed by analyzing the extent of implant enhancement seen on MR imaging. RESULTS Of 21 patients undergoing gadolinium-DTPA T1-weighted MR imaging 2 to 7 weeks after HA placement, 15 had enhancement limited to the implant rim (Grade I or less). Five patients had peripheral foci of enhancement (Grade II), and one patient had foci of enhancement extending to the center of the implant (Grade III). MR images obtained 9 to 15 weeks after HA insertion in all 14 patients had some degree of central enhancement (Grade III) and 11 had homogeneous enhancement throughout the implant (Grade IV or V). Seven patients in the homogeneous group were believed to have particularly intense enhancement patterns (Grade V). Of the 10 patients undergoing MR imaging from 31 to 69 weeks after surgery, 5 had Grade III enhancement and 5 had Grade IV enhancement. CONCLUSIONS: This study demonstrated consistent central HA orbital implant enhancement on MR imaging in the 9- to 15 week group and the >31-week postoperative group. HA orbital implant drilling and peg placement should be performed after central vascularization of the spherical implant has occurred. The results of this study support the principle of performing orbital implant drilling and peg placement at least 5 to 6 months after HA implant insertion. PMID- 12544794 TI - Complications of supramid orbital implants. AB - PURPOSE: To determine the incidence and risk factors of complications associated with Supramid orbital implants. METHODS: A retrospective chart review was performed to document complications in a series of 41 patients receiving Supramid implants during orbital reconstruction or fracture repair. Implant size, duration of time between trauma and implant insertion, and history of prior orbital surgery were examined as possible risk factors for the development of complications. RESULTS: Four patients had hemorrhage within the implant capsule. An orbital abscess developed in a single patient. All but one complication appeared 7.8 to 10 years after implant insertion. Patients who had complications had a longer duration of time between trauma and implant insertion than those in whom complications did not occur (P =0.0019). Complications were more frequent among patients with larger (>600 mm2) implants and a history of orbital surgery. CONCLUSIONS: Spontaneous infection or hemorrhage may occur within the capsules of Supramid orbital implants, even many years after surgery. The insertion of larger implants in the late repair of extensive bony orbital defects may predispose patients to these complications. Implant removal and marsupialization of the implant capsule to the maxillary sinus appears to be curative without causing significant postoperative enophthalmos. PMID- 12544795 TI - Echography as a useful adjunct in the diagnosis of orbital solitary fibrous tumor. AB - PURPOSE: To describe the echographic features of solitary fibrous tumor of the orbit. METHODS: A retrospective review of 3 patients with solitary fibrous tumor of the orbit and a literature review were performed. RESULTS: Three patients with orbital solitary fibrous tumor were studied with standardized echography and 3 other cases of orbital solitary fibrous tumor with ultrasound descriptions were identified in a literature review. Consistent echographic features of the 6 orbital solitary fibrous tumors included firmness, low to medium internal reflectivity, regular internal structure, moderate sound attenuation, and some degree of vascularity. CONCLUSIONS: Standardized echography is a useful adjunct in the evaluation of orbital solitary fibrous tumors and can help differentiate these tumors from other orbital lesions. PMID- 12544796 TI - Tegaderm transparent dressing (3M) for the treatment of chronic exposure keratopathy. AB - We report an effective alternative treatment of profound chronic exposure keratopathy in a proptotic eye due to intraorbital extension of basal cell carcinoma. The corneal surface gradually reepithelialized in a 6-week period after the first application of Tegaderm transparent dressing (3M, St. Paul, MN, U.S.A.) with instillation of antibiotic ointment on the ocular surface. The patient has been comfortable with an intact epithelial surface, a vascularized cornea, and nonirritated surrounding skin after changing the Tegaderm dressing daily for 15 months. There was no evidence of recurrent corneal ulceration or infiltrations. We conclude that Tegaderm represents a useful treatment of exposure keratopathy due to chronic proptosis. PMID- 12544798 TI - Severe congenital horizontal tarsal kink without levator aponeurosis disinsertion. AB - A newborn presented with congenital severe tarsal kink of the upper eyelids not caused by levator aponeurosis disinsertion. A bilateral anterior lamellar repositioning procedure was performed to correct the tarsal kink. Levator aponeurosis disinsertion was not observed as an intraoperative finding. The tarsal kink disappeared, and eyelid positions remained stable during a follow-up period of 1 year. Our case indicates that severe congenital tarsal kink may develop without levator aponeurosis disinsertion, and anterior lamellar repositioning technique is a simple and effective method of treatment. PMID- 12544797 TI - Occult orbital neuroblastoma detected after administration of an antitumor vaccine. AB - A 6-year-old girl with neuroblastoma developed swelling and erythema of her right upper eyelid following administration of an interleukin-2 and lymphotactin gene modified allogeneic neuroblastoma cell vaccine. Computed tomography demonstrated a cystic lesion in the subperiosteal space. A biopsy of the mass showed necrotic neuroblastoma with minimal associated inflammation. To our knowledge, this case represents the first description of occult orbital metastases in a patient with neuroblastoma detected after administration of an antitumor vaccine. PMID- 12544799 TI - Tarsal kink syndrome associated with congenital corneal ulcer. AB - We describe a case of unilateral upper eyelid horizontal tarsal kink with congenital corneal ulceration. The surgical repair, by means of a simple resection of orbicularis and skin, is presented as a simple technique to correct the congenital anomaly of tarsal kink. Congenital tarsal kink should be included in the differential diagnosis of congenital corneal opacity and ulceration. Early recognition and surgical intervention reduces corneal scarring. PMID- 12544800 TI - Diplopia following porous polyethylene orbital rim onlay implant. AB - An 81-year-old man with ocular irritation associated with lower eyelid retraction, horizontal laxity of the lower eyelids, and hypoplastic inferior orbital rims underwent bilateral placement of porous polyethylene orbital rim onlay implants. Two weeks after surgery, he developed vertical binocular diplopia on downgaze. Examination of extraocular motility demonstrated limited infraduction OD. Surgical exploration revealed scarring in the anterior orbit between the inferior rectus pulley and the orbital implant. The orbital implant was found to lie higher than the inferior orbital rim. After surgical lysis of the scar and reduction of the vertical height of the implant, the patient's diplopia resolved. Orbital connective tissues critical to ocular motility may be abnormally superficial in orbital rim hypoplasia. Onlay grafts must be carefully placed so that they do not interfere with these tissues. PMID- 12544801 TI - Re: "Combined flap repair of moderate lower eyelid defects". PMID- 12544802 TI - Re: "Combined flap repair of moderate lower eyelid defects". PMID- 12544803 TI - Management of coincident cataract and uveitis. AB - Cataract surgery in a patient with uveitis is more complex than senile cataract extraction, because it involves multiple considerations related to the cause of uveitis, prospects of visual rehabilitation, appropriate surgical timing and technique, and the type and material of the intraocular lens used. Establishing the diagnosis, thorough eye examination, careful patient selection and meticulous control of perioperative inflammation are key elements to a successful visual outcome. Our aims in this article are to review the literature on this subject over the past year and highlight the behavior of intraocular lenses of various biomaterials in the uveitic eye. In addition, we also reemphasize the idea of a model of zero tolerance to intraocular inflammation to minimize the incidence of irreversible damage to ocular structures essential to good vision. PMID- 12544804 TI - Techniques for managing common complications of cataract surgery. AB - Cataract surgery has evolved dramatically over the last two decades, largely as a result of technological advances. As a result, visual outcomes and patient convalescence have improved significantly. A second consequence of increased instrumentation and technology, however, is increased complexity of cataract surgery and the advent of complications unique to these advances. Cataract surgeons must be aware and capable of managing the many possible adverse events that can occur during cataract surgery. This review identifies many of the common complications of cataract surgery, describes methods to avoid these complications and discusses techniques to address complications that do occur. It is the ability to avoid, quickly identify, and properly manage complications that defines the accomplished cataract surgeon. PMID- 12544805 TI - White Star technology. AB - White Star micropulse technology is a software modification that allows extremely short bursts of ultrasound energy. Studies have shown that this decreases wound heat build-up with the retained efficiency of continuous ultrasound. Decreased energy utilization with improved corneal function and improved nuclear fragment followability appear to be additional benefits. PMID- 12544806 TI - Refractive lens exchange with a multifocal intraocular lens. AB - Refractive lens exchange with a multifocal intraocular lens is becoming a more popular method of refractive surgery in the presbyopic patient. The limitations of keratorefractive surgery have led to a resurgence of lens exchange surgery for patients with prescriptions outside the limits of corneal refractive procedures, in addition to patients with routine refractive errors requesting a surgical procedure to achieve emmetropia and also address presbyopia. Side effects of multifocal technology including unwanted photic phenomena and deterioration in contrast sensitivity are being further defined and evaluated to better assess the effects of these intraocular lenses on functional vision and patient satisfaction. Attention to detail in regards to proper patient selection, preoperative measurements, intraoperative technique, and postoperative management will ultimately result in excellent outcomes and improved patient acceptance of this effective technique. PMID- 12544807 TI - AdvanTec Legacy System and the NeoSoniX handpiece. AB - AdvanTec Legacy System and the NeoSoniX handpiece (Alcon Surgical, Fort Worth, Texas) are improvements to the Legacy System that allow power modulation and customization of surgical technique in phacoemulsification surgery. AdvanTec technology allows power modulation in terms of occlusion power management, global linear flow, mode sequencing, and visco mode. The NeoSoniX handpiece combines sonic oscillations with linear ultrasound to reduce thermal energy during nucleus removal. The AdvanTec System allows lower power, less delivered energy, a lower risk of thermal burns, less chatter, and improved followability during phacoemulsification. These improvements translate into successful clinical outcomes. PMID- 12544808 TI - Treating astigmatism at the time of cataract surgery. AB - An increasingly important goal of modern cataract and implant surgery is to obtain the most desirable refractive outcome for our patients, and in so doing, decrease their dependence upon spectacle correction. Refined spherical outcomes are now achieved by directing close attention to biometry technique, through the use of third generation intraocular lens (IOL) power formulae, and refinements in technology such as partial coherence interferometry. Similarly, improvements in astigmatic outcomes are now possible by focusing upon and obtaining more accurate preoperative cylinder measurements, careful surgical planning particularly in regard to incision(s) design, use of toric IOLs, and employing advanced postoperative enhancement techniques. Because of these refinements in technique and technology, the field of cataract and implant surgery has emerged as a true and recognized form of refractive surgery. PMID- 12544809 TI - Managing cystoid macular edema after cataract surgery. AB - Cystoid macular edema (CME) is a common problem after cataract extraction. It can occur after uncomplicated surgery in patients with otherwise healthy eyes, after complicated surgery, or after surgery in patients with ocular diseases such as uveitis or diabetic retinopathy. Usually vision loss from cystoid macular edema is temporary and responds to treatment with topical anti-inflammatory medications. However, some cases respond poorly to conservative treatment and may develop permanent visual loss. A review of the medical literature was performed for all articles published in English between August 1, 2001 and July 31, 2002 on the topic of cystoid macular edema after cataract surgery. The authors selected nine articles that were most relevant to the practicing ophthalmologist for inclusion in this review. Topics of interest included vitreous loss, retained lens fragments, diabetes, uveitis, retinitis pigmentosa, ocular hypotensive lipids, internal limiting membrane peeling, and intravitreal triamcinolone injection. PMID- 12544810 TI - Cataract surgery in patients with prior refractive surgery. AB - As the number and types of keratorefractive procedures increase and as the baby boomer population moves into the "cataractous decades," the number of patients requiring cataract surgery following refractive surgery grows larger each year. While technological advances in surgical instrumentation and intraocular lens (IOL) design allow us to perform cleaner, faster, and more reliable cataract extractions, the ultimate postoperative refraction depends primarily on calculations performed before surgery. Third-generation IOL formulas ( Haigis, Hoffer Q, Holladay 2, or SRK/T) provide outstanding accuracy when used for eyes with physiologic, prolate corneas. In addition, most instruments used today for measuring corneal curvature and power were designed before the era of refractive surgery. These formulas and instruments make assumptions about the anatomy and refractive properties of the cornea that are no longer valid following most keratorefractive procedures. These breakdowns in IOL calculation often result in a "refractive surprise" after cataract surgery, which may require subsequent surgical correction. This article examines recent publications of modeling studies of various methods for estimating effective K values for IOL calculation, cataract surgery case series following refractive surgery, new corneal topography technologies and methods for correcting "refractive surprises" postoperatively. PMID- 12544811 TI - Introduction and overview of the conceptual framework for a national quality measurement and reporting system. AB - BACKGROUND: Little systematic information is currently available on the level of quality of health care received in the United States. Most studies find significant deficits in quality, but without systematic information on quality at all levels in the health care system, substantial improvements in care delivery are unlikely. OBJECTIVES: The Strategic Framework Board (SFB) was established in 1999 to (1) design a strategy for a national quality measurement and reporting system (NQMRS), (2) articulate the guiding principles and priorities for a national system, and (3) identify potential barriers to successful implementation and propose possible solutions. RESEARCH DESIGN: The SFB consisted of nine members selected for their breadth of research and experience in quality. The work was conducted as an expert deliberation, including face-to-face meetings, regular conference calls, some literature reviews on specific topics, drafting and discussion of papers, and presentation of observations and recommendations to the Board of the National Forum for Health Care Quality Measurement and Reporting. FINDINGS: The SFB developed a conceptual framework to organize the design of the NQMRS. The major components include: statement of purpose, selection of national goals, establishment of the NQMRS on a scientific foundation, selection of a common set of measures and effective reporting strategies, facilitation of local adoption of the system, and development of the capacity for quality improvement. CONCLUSIONS: An NQMRS is essential for the nation to make improvements in quality. The work here presents a first step in designing such a system. Considerable work on making these strategies operational remains to be undertaken. PMID- 12544812 TI - An evidence-based national quality measurement and reporting system. AB - BACKGROUND: Evidence-based medicine and evidence-based guidelines are widely supported among physicians and other health professionals, although practice may still fall far short of the ideas contained in the concept. Insufficient attention has been given to using the concepts of evidence-based practice in the selection of measures, diffusion of innovations, and adoption of quality improvement (QI) techniques. OBJECTIVES: To describe in greater detail how evidence serves as a foundation for a national quality measurement and reporting system (NQMRS) and to consider the different types of evidence that will be used. The role of the NQMRS in adding to the evidence base is explored. RESEARCH DESIGN: A descriptive, conceptual discussion. RESULTS: Many different types of evidence are available for use in the NQMRS. The NQMRS could add substantial new evidence on how well different approaches to quality measurement, reporting, and improvement work and also expand the evidence base on the effectiveness of clinical interventions for which efficacy has been established. CONCLUSIONS: An evidence-based NQMRS will require substantial leadership from government and private entities. The gains from a greater focus on evidence are expected to be found in accelerating QI. Reliance on evidence-based approaches should not be used to stifle innovation. PMID- 12544813 TI - Establishing national goals for quality improvement. AB - BACKGROUND: For a number of reasons, it has been difficult to garner public interest in the need to improve the quality of care delivered nationally. One possible reason for this is that the concept of quality has different meanings for different stakeholders. To make the problems of suboptimal quality more concrete and potential solutions more compelling, the Strategic Framework Board (SFB) recommended developing a set of national goals for quality improvement. OBJECTIVES: To describe the criteria by which national goals should be selected, illustrate the analytic methods that should be used to support the development of such goals, and describe and illustrate a process by which national goals could be formulated. RESEARCH DESIGN: Targeted review of literature and discussions among members of the SFB. FINDINGS: National goals have played a key role in making progress under the Healthy People 2000 and Health People 2010 initiatives. The recommended process will involve assembling key evidence as well as engaging in a consensus process. CONCLUSIONS: Developing a set of national goals for quality improvement is a key activity for a national quality measurement and reporting system to undertake. The steps outlined here represent a feasible and productive method for accomplishing this objective. PMID- 12544814 TI - Connections between quality measurement and improvement. AB - BACKGROUND: Measurement is necessary but not sufficient for quality improvement. Because the purpose of the national quality measurement and reporting system (NQMRS) is to improve quality, a discussion of the link between measurement and improvement is critical for ensuring an appropriate system design. OBJECTIVES: To classify approaches to the use of measurement in improvement into two different- although linked and potentially synergistic--agendas, or "pathways." To discuss the barriers encountered in each of these pathways and identify steps needed to motivate improvement in both pathways. RESEARCH DESIGN: Descriptive, conceptual discussion. FINDINGS: The barriers to the use of information to motivate change include, in Pathway I (selection), the lack of skill, knowledge, and motivation on the part of those who could drive change by using data to choose from among competing providers, and, in Pathway II (change in care delivery), the deficiencies in organizational and professional capacity in health care to lead change and improvement itself. CONCLUSIONS: Neither the dynamics of selection nor the dynamics of improvement work reliably today. The barriers are not just in the lack of uniform, simple, and reliable measurements, they also include a lack of capacity among the organizations and individuals acting on both pathways. PMID- 12544815 TI - Selecting common measures of quality and system performance. AB - BACKGROUND: The President's Commission on Consumer Protection and Quality in the Health Care Industry recommended that a common set of quality measures be developed for the nation. The results of such common measures will be used to ensure accountability, select providers, and improve quality. Simultaneous consideration of top-down and bottom-up design requirements are likely to produce a set of measures that will serve policy and front-line information needs. OBJECTIVES: To articulate the criteria and process by which common measures should be selected and to illustrate the results of applying this approach in one clinical area. DESIGN: Discussions among the members of the Strategic Framework Board, development of a clinical logic model for acute myocardial infarction (AMI), and application of the criteria to existing quality measures for AMI. FINDINGS: Measures should: (1) be linked to a national goal, (2) have a clear and compelling use, (3) be parsimonious, (4) not impose undue burden on those providing data, (5) help providers improve care delivery, (6) help stakeholders make more informed decisions, and (7) balance the need for continuous improvement with the stability needed to track progress over time. The use of a clinical logic diagram highlights the importance of selecting measures related to primary and secondary prevention in reducing deaths from heart disease. The resulting measures are useful on the front lines of medicine as well as by consumers and purchasers. CONCLUSIONS: Focusing attention on the information necessary to stimulate progress on national goals provides a compelling framework for the choice of a common set of measures. PMID- 12544816 TI - Using performance measurement to drive improvement: a road map for change. AB - BACKGROUND: Performance measures and reporting have not been adopted throughout the US health care system despite their central role in encouraging increased participation by consumers in decision-making. Understanding whether the failure of measurement and reporting to diffuse throughout the health system can be overcome is critical for determining future policy in this area. OBJECTIVES: To create a conceptual framework for analyzing the current rate of adoption and evaluating alternatives for accelerating adoption, and to recommend a set of concrete steps that can be taken to increase the use of performance measurement and reporting. RESEARCH DESIGN: Review of three theoretic models (Rogers, Prochaska/DiClemente, Gladwell), examination of the literature on previous experiences with quality measurement and reporting, and interviews with select stakeholders. FINDINGS: The three theoretic models provide a valuable framework for understanding why the use of performance measures is stalled ("the circle of unaccountability") and for generating ideas about concrete steps that could be taken to accelerate adoption. Six steps are recommended: (1) raise public awareness, (2) redesign measures and reports, (3) make the delivery of information timely, (4) require public reporting, (5) develop and implement systems to reward quality, and (6) actively court leaders. CONCLUSIONS: The recommended six steps are interconnected; action on all will be required to drive significant acceleration in rates of adoption of performance measurement and reporting. Leadership and coordination are necessary to ensure these steps are taken and that they work in concert with one another. PMID- 12544817 TI - Engaging health care consumers to improve the quality of care. AB - BACKGROUND: Researchers and policy makers have focused on informed choice as the primary role for consumers in improving care. Consumers may play two other roles in improving quality--by being active participants in their care (coproducer role) and by evaluating the care they receive (evaluator role). Enhancing the roles consumers are able to play in the health care system may significantly increase their influence on quality. OBJECTIVES: To examine the evidence available on the contribution of consumers to quality through the coproducer and evaluator roles. RESEARCH DESIGN: Conceptual framework and review of the literature. FINDINGS: Patients who engage in collaborative care, shared decision making with their providers, and chronic disease self-management have improved health outcomes. Training patients with chronic diseases to self-manage their disease increases functioning, reduces pain, and decreases costs. Developing the evaluator role will support and increase the effectiveness of the other two roles the informed choice role and the coproducer role. CONCLUSIONS: Only the informed choice role is being actively promoted for consumers. Increasing the coproducer role would require system and provider change, as well as an increase in consumers' skills and knowledge and a change in their understanding of their appropriate role. Harnessing the power of consumers to create change will depend on the degree to which all these roles are encouraged and supported. PMID- 12544818 TI - Information system concepts for quality measurement. AB - BACKGROUND: Health care information systems in use today frequently fall short of what is needed to meet the demands for data and reporting on performance. Many observers believe substantial improvements in information systems will be necessary if the potential of a national quality measurement and reporting system (NQMRS) is to be realized. A shared vision will facilitate progress in improving information systems. OBJECTIVES: To articulate a set of guiding principles and operational steps for the development of functional information systems in health care. RESEARCH DESIGN: Experience in building such systems for one health care delivery system was used to develop an approach. This was discussed with Strategic Framework Board members and integrated with other considerations for going from a local system to one that could accumulate information for national purposes. FINDINGS: The key elements of a functional information system include provisions that (1) data should be collected once, (2) aggregation of data for higher-level reports should be anticipated, (3) issues related to privacy and confidentiality must be addressed, and (4) measurement systems should include an audit standard. A seven-step process for developing a functional information system is outlined. CONCLUSIONS: A shared national measurement framework is essential because the data systems that health care delivery organizations use are not static. A long-term vision can guide the growth of a data system over time. An NQMRS can be the vehicle that provides the needed vision. PMID- 12544819 TI - A research agenda to advance quality measurement and improvement. AB - BACKGROUND: In developing a conceptual framework for the design of a national quality measurement and reporting system (NQMRS), the Strategic Framework Board (SFB) recommends that such a system be built on a strong evidence base. OBJECTIVES: To identify critical gaps in the evidence needed for a fully functional NQMRS and to recommend a starting point for the development of a research agenda. RESEARCH DESIGN: Selective review of literature in quality of care measurement and reporting and identification of strategic issues that must be addressed. FINDINGS: There is some limited evidence that measurement and reporting can improve quality. Substantial advances have been made in the science of measurement and reporting but important gaps remain, specifically in (1) measurement methods and tools, (2) uses of quality performance data, (3) organizational and cultural factors, (4) information and informatics, and (5) impact evaluation/research. CONCLUSIONS: To achieve a sustainable research agenda, three strategic issues will have to be addressed: (1) the policy rationale for the research agenda, (2) adequate levels of public-sector funding, and (3) sustainability in a rapidly changing environment. PMID- 12544820 TI - Putting the ideas into practice. PMID- 12544821 TI - Opioid-induced hyperalgesia: abnormal or normal pain? PMID- 12544822 TI - Responses to sympathomimetics in rat sensory neurones after nerve transection. AB - Noradrenaline activation of sensory somata that project in damaged peripheral nerves has been postulated to underlie sympathetically-mediated pain. Intracellular recordings from some neurones with myelinated axons in acutely isolated rat dorsal root ganglia showed small prolonged depolarizations to brief applications of 0.1-5 mM noradrenaline whether or not the spinal nerve had been transected. Similar responses were evoked to noradrenaline when phentolamine was present, and also to 1-5 mM catechol, but not 1 mM clonidine, implying the responses were not adrenoceptor-mediated. In extracellular recordings from similar preparations after sciatic transection, many spontaneously active myelinated dorsal root axons were excited by noradrenaline and other sympathomimetics. Silent axons in injured or control ganglia did not respond. Thus, non-specific depolarizations may activate neurones that are hyperexcitable after a lesion but activation of neuronal alpha-adrenoceptors by sympathetically released noradrenaline seems unlikely. PMID- 12544823 TI - Involvement of CLOCK:BMAL1 heterodimer in serum-responsive mPer1 induction. AB - A rapid induction of mouse period1 (mPer1) gene expression is supposed to be critical in the clock gene regulation, especially in the phase resetting of the clock, but its molecular mechanism is poorly understood. Based on the previous finding that the process does not involve de novo synthesis of proteins, we postulated the involvement of CLOCK:BMAL1 heterodimer, a positive regulator of circadian oscillator, in the rapid induction of mPer1 transcription. To test this hypothesis, we utilized CLOCKdelta19, a dominant-negative mutant, to suppress the function of CLOCK:BMAL1 in vitro. Serum-evoked rapid increases of mPer1 mRNA expression and promoter activity were significantly blunted when CLOCK:BMAL1 function was interfered with. Furthermore, DNA binding activity of CLOCK:BMAL1 heterodimer to five E-boxes of mPer1 promoter markedly increased shortly after serum shock. Taken together, these results suggest that CLOCK:BMAL1 heterodimer is not only a core component of negative feedback loop driving circadian oscillator, but also involved in the rapid induction of mPer1during phase resetting of the clock. PMID- 12544824 TI - Pallidotomy and incidental sequence learning in Parkinson's disease. AB - Converging evidence from animal research and human brain imaging studies, points to an important role of cortical-striatal motor circuitry in the incidental learning of serial order information. To date, attempts to address this role through the study of patients with striatal disorder have proved inconclusive. The present study examined the impact of a therapeutic lesion of the globus pallidus in patients with Parkinson's disease. The lesion, which blocks a primary output of the putamen to the motor cortices, eliminated incidental learning relative both to controls and unoperated patients. The finding offers support for models proposing that context detection within the striatum is a central process in serial order learning. An unexpected effect of the lesion was to significantly reduce the response time to random stimuli relative to an ordered series, the opposite of the normal pattern. It is speculated that this may reflect an unconscious alerting response to novelty, a process suggested to involve the ventral striatum and its cortical targets. Research on Parkinson's disease patients undergoing functional basal ganglia surgery may shed further light on the mechanisms and neuronal substrate of serial order learning in humans. PMID- 12544825 TI - A novel putative M9.2 isoform of V-ATPase expressed in the nervous system. AB - We have identified a cDNA encoding a novel putative neuron-specific isoform of vacuolar proton-translocating ATPase (V-ATPase), NM9.2, from rat and mouse. Sequence analysis revealed that NM9.2 conserved similar characteristic amino acid sequences with 60-70% identity to M9.2 previously isolated from V-ATPase in chromaffin granules. Using Northern blot analysis, NM9.2 mRNA was specifically detected in the brain, whereas M9.2 mRNA was widely expressed in various tissues. In situ hybridization showed that NM9.2 gene expression was restricted mainly to neuronal cells and consistent with that of the a1/Ac116 subunit of V-ATPase. NM9.2 is a putative neuronal isoform of the 9.2 kDa subunit in V-ATPase. PMID- 12544826 TI - The septin protein Nedd5 associates with both the exocyst complex and microtubules and disruption of its GTPase activity promotes aberrant neurite sprouting in PC12 cells. AB - Nedd5 is a septin protein enriched in brain and associates with the exocyst complex, a protein complex required for neurite outgrowth in neuroendocrine PC12 cells. In this study, we further investigate the association between Nedd5 and the exocyst complex as well as the role of Nedd5 in neurite outgrowth in differentiating PC12 cells. The endogenous Nedd5 is enriched at the perinuclear region in undifferentiated PC12 cells and radiates outward, from the perinuclear region toward the growth cone, upon NGF-induced PC12 neuronal differentiation. Nedd5, as well as other septin proteins, co-immunoprecipitates with the exocyst complex and tubulin from rat brain lysate. Interestingly, the over-expression of a GTPase-defective Nedd5 mutant promotes aberrant neurite sprouting in PC12 cells. These results demonstrate that Nedd5 and other septin proteins are associated with both the exocyst complex and microtubules and uncover a putative role for the Nedd5 GTPase activity in neurite outgrowth. Taken together, these findings suggest that Nedd5 may be required for polarized neurite outgrowth, perhaps, by facilitating the exocyst complex function during neuronal differentiation. PMID- 12544827 TI - Effects of implantation site of dead stem cells in rats with stroke damage. AB - Searching for valid control grafts, we assessed the performance of rats subjected to middle cerebral artery occlusion (MCAO) and grafted with freeze-thawed dead stem cells into sites previously used for active grafts (ipsilateral and contralateral striatum and ventricle) on bilateral asymmetry and water maze tests. We expected to find that sham grafted groups had impairments equivalent to those of MCAO-only controls, relative to intact controls. This proved to be the case for contralateral and intraventricular grafts, and for asymmetry in rats with ipsilateral grafts. However, spatial learning was substantially impaired and lesion volume was increased by 55% with ipsilateral dead cell grafts. Exacerbation of stroke effects indicates potential hazards in the use of dead cells for sham grafts. PMID- 12544828 TI - Progesterone withdrawal increases the anxiolytic actions of gaboxadol: role of alpha4betadelta GABA(A) receptors. AB - Hippocampal alpha4betadelta GABA(A) receptors (GABA(A)-R) are increased following progesterone withdrawal (PWD) in a rodent model of premenstrual anxiety. This alpha4betadelta receptor isoform uniquely responds to the GABA agonist gaboxadol (THIP) with a maximum current greater than that gated by GABA, and is potentiated more by pentobarbital than are other GABA(A)-R. We therefore investigated the anxiolytic effects of these drugs using the elevated plus maze. Gaboxadol (1.25 mg/kg) was markedly more anxiolytic in animals undergoing PWD than in controls. Pentobarbital (10 mg/kg) also produced a greater anxiolytic effect during PWD. These results suggest that the pharmacological properties of alpha4betadelta GABA(A)-R following PWD are evident behaviorally. Alterations in the alpha4betadelta GABA(A)-R population may have implications for the etiology and treatment of premenstrual syndrome. PMID- 12544829 TI - Differences in dopaminergic neuroprotective effects of estrogen during estrous cycle. AB - Previous studies suggest that estrogen treatment protects nigrostriatal dopaminergic neurons, but have not examined whether the changes in estrogen levels during estrous cycle can influence the susceptibility of these neurons to neurotoxins. Here we show that the loss of dopaminergic neurons in the substantia nigra was greater in animals lesioned at diestrus (low estrogen) using 6 hydroxydopamine or buffered iron chloride, when compared with animals lesioned at proestrus (high estrogen). Lesioning at diestrus with 6-hydroxydopamine reduced the striatal dopamine content, whereas the dopamine content was preserved in animals lesioned at proestrus. The density of the dopamine transporter, upon which 6-hydroxydopamine toxicity is dependent, was lower when circulating estrogen was high. These results thus support a neuroprotectory role for estrogen. PMID- 12544830 TI - Brain activity during expectancy of emotional stimuli: an fMRI study. AB - We studied the neural activation associated with the expectancy of emotional stimuli using whole brain fMRI. Fifteen healthy subjects underwent fMRI scanning during which they performed a warned reaction task using emotional pictures carrying pleasant, unpleasant, or neutral content. The task involved an expected or unexpected condition. Data were analyzed by comparing the images acquired under the different conditions. In the expected condition, compared with the unexpected condition, significant activation was observed in the medial, inferior and dorsolateral prefrontal cortex. Whereas the expectancy of pleasant stimuli produced activation in the left dorsolateral and left medial prefrontal cortex as well as in the right cerebellum, the expectancy of unpleasant stimuli produced activation in the right inferior and right medial prefrontal cortex, the right amygdala, the left anterior cingulate cortex, and bilaterally in the visual cortex. These results suggest that the expectancy of emotional stimuli is mediated by the prefrontal area including the medial, inferior, and dorsolateral prefrontal cortex. Furthermore, our data suggest that left frontal activation is associated with the expectancy of pleasant stimuli and that right frontal activation is associated with the expectancy of unpleasant stimuli. Finally, our findings suggest that the amygdala and anterior cingulate cortex may play an important role in the expectancy of unpleasant stimuli and that the input of this negative information is modulated by these specific brain areas. PMID- 12544831 TI - Post-lesion administration of 5-HT1A receptor agonist 8-OH-DPAT protects cholinergic nucleus basalis neurons against NMDA excitotoxicity. AB - Recent evidence indicates that serotonin (5-HT)1A receptor agonists may abrogate excitotoxic brain damage. We investigated whether a single i.p. injection of the 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT), at a dose of 2.5 mg/kg, protects cholinergic neurons of the rat magnocellular nucleus basalis (MBN) against NMDA excitotoxicity when administered at post-injury intervals ranging from 6 to 96 h. Drug effects on passive avoidance learning and on the density of cortical cholinergic innervation, a measure of neuronal survival in the damaged MBN, were analyzed. Our results demonstrate that 8-OH DPAT, when administered up to 24 h post-lesion, significantly attenuates both behavioral and neuroanatomical consequences of NMDA excitotoxicity on cholinergic MBN neurons; and support the hypothesis that 5-HT1A receptor agonists may interfere with delayed neuronal death in vivo that is of significance in the pharmacological treatment of neurological disorders associated with excitotoxic neuronal damage. PMID- 12544832 TI - Effect of amyloid precursor protein 17mer peptide on microtubule structure and tau protein hyperphosphorylation in hippocampal neurons of experimental diabetic mice. AB - The objective of this study was to investigate whether microtubular structure changes and tau protein hyperphosphorylation exist in hippocampal neurons of experimental diabetic mice, and to study the effect of amyloid precursor protein 17mer peptide. The results showed that the microtubules of hippocampal neurons of diabetic mice manifested prominent signs of fragmentation and dissolution, tau protein is hyperphosphorylated at Ser 199/Thr 202 sites, enzymes related to the phosphorylation and dephosphorylation of tau protein were diminished. The administration of amyloid precursor protein 17mer peptide could ameliorate the foregoing changes in diabetic mice. These results indicated that protein synthesis in the brain tissue of diabetic mice decreased. Amyloid precursor protein 17mer peptide acted as a neuroprotective agent that globally alleviates the disturbances due to impaired energy metabolism in diabetic mice. PMID- 12544833 TI - Fronto-parietal evoked potential synchronization is increased during mental rotation. AB - We used steady state visually evoked potential event related partial coherence (SSVEP-ERPC) to examine the SSVEP synchronization between brain regions while 22 males undertook a sequential version of the Shepard and Metzler mental rotation task. Compared to the 60 degrees rotation, the 180 degrees rotation was associated with increased synchronization between bilateral prefrontal and parieto-occipital sites, between left frontal and right parietal sites and between bilateral parietal and occipital sites. We suggest that the increased synchronization between prefrontal and parieto-occipital regions may be associated with the working memory components of the task, while the left frontal to right parietal synchronization may represent the increased interaction between these regions thought to occur in a variety of visuo-motor tasks. PMID- 12544834 TI - Nucleus-specific alteration of raphe neurons in human neurodegenerative disorders. AB - Neurodegenerative diseases share symptoms suggested to be related to the serotonergic system. To evaluate the involvement of serotonergic raphe nuclei, we compared the percentage of neurons synthesizing serotonin in the nucleus centralis superior (NCS), raphe obscurus and pallidus (NROP) in Alzheimer's disease (AD), progressive supranuclear palsy (PSP), Parkinson's disease (PD), multiple system atrophy (MSA), and control brains. We used immunohistochemistry for tryptophan hydroxylase (TpOH), phosphorylated tau, and alpha-synuclein. We observed a significant decrease in the NCS in the NROP in AD, but a significant increase in PSP and MSA. Cytoskeletal pathology was present in the NCS and NROP to a variable degree. We conclude that there is disease- and nucleus-specific alteration of serotonin synthesis in the raphe. PMID- 12544835 TI - Trophic support of mouse inner ear by neural stem cell transplantation. AB - In the auditory system, efforts to reduce degeneration of spiral ganglion neurons have the immediate objective of improving clinical benefits of cochlear implants, which are small devices designed to stimulate spiral ganglion neurons electronically. Recent studies have indicated several neurotrophins can enhance survival of spiral ganglion neurons. However, the strategy for application of neurotrophins in inner ear is still a matter of debate. In this study, we examined the potential of cell therapy as a strategy for application of neurotrophins in the inner ear. Neural stem cells obtained from green fluorescent protein-transgenic mice were used as donor cells. Medium containing neural stem cells was injected into mouse inner ear. Histological analysis 4 weeks later revealed that transplant-derived cells survived in inner ear and that most transplant-derived cells in the cochlea had differentiated into glial cells. Moreover, expression of glial cell line-derived neurotrophic factor and brain derived neurotrophic factor was observed in transplant-derived cells. These findings indicate that transplantation of neural stem cells can be a useful strategy for application of neurotrophins in inner ear. PMID- 12544836 TI - Nigrothalamostriatal and nigrothalamocortical pathways via the ventrolateral parafascicular nucleus. AB - The present tract-tracing study in the rat indicated that neurons in the ventrolateral part of the parafascicular thalamic nucleus (PF), where nigral fibers from the dorsolateral part of the substantia nigra pars reticulata (SNr) terminated, sent their axons to the ventrolateral part of the striatum as well as to the rostrolateral part of the lateral agranular cortex (AGl). We further demonstrated that symmetrical synaptic contacts were made between these nigral axons and striatum- or AGl-projecting PF neurons. Since the dorsolateral part of the SNr, ventrolateral part of the striatum and rostrolateral part of the AGl are responsible regions for orofacial behaviors, the nigrothalamostriatal and nigrothalamo-cortical pathways via the ventrolateral part of the PF may play a role in the control of orofacial motor function. PMID- 12544837 TI - DC11: a novel monoclonal antibody revealing Alzheimer's disease-specific tau epitope. AB - Using tau protein extracts from Alzheimer's disease (AD) brain tissue, we generated a monoclonal antibody (mAb DC11) which decorated neurofibrillary pathology in brain derived from AD patients on immunohistochemistry, and which lacked reactivity with healthy brain tissue. The same pattern of DC11 specificity was observed on Western blot. The main constituent of structures decorated by DC11 is microtubule-associated protein tau. In Western blot, DC11 recognized neither native healthy tau nor its full length recombinant counterpart. However, the mAb showed strong immunoreactivity with truncated tau (residues tau151-421), thus indicating the requirement for a conformational epitope. Importantly, the DC11 epitope was phosphorylation independent. The immunochemical parameters of mAb show that DC11 could represent a novel structural probe with the specificity for conformation of pathological tau present in AD brains. PMID- 12544838 TI - Dopamine D3 receptor ligands show place conditioning effect but do not influence cocaine-induced place preference. AB - The importance of dopamine D3 receptors in reward related processes, especially in cocaine addiction, has been investigated extensively. However, in the reported studies a combination of different experimental conditions and different ligands have been used which renders the interpretation and comparison of the diverse results extremely difficult. Here, we report one comparative study investigating a wide range of dopamine D3 receptor ligands in one model of cocaine abuse: the place conditioning paradigm in rats. Of the antagonists tested, the moderately D3 selective nafadotride and the more selective SB-277011 did not produce any place conditioning effect while U-99194A caused place-preference. The most D3 selective agonist PD-128907, the less selective 7-OH-DPAT and the moderately selective partial agonist BP-897 all caused significant place aversion. None of the compounds influenced the cocaine-induced place preference. Results suggest the D3 preferring agonists could affect the reward mechanisms of the brain, however, modulation of D3 receptor function does not appear to be a significant mechanism for modifying the place conditioning effect of cocaine. PMID- 12544839 TI - Task-dependent visual coding of sound position in visuospatial neglect patients. AB - Recent neurophysiological evidence has shown that sound position can be coded in multiple frames of reference in the animal brain (i.e. head-centred, eye-centred, or intermediate head/eye centred). Here, we provide evidence for multiple coding of sound positions in humans, by studying pointing to sounds in 14 right brain damaged (RBD) patients with or without visual neglect (a visuospatial neurological disturbance typically affecting contralesional space). Patients were asked to indicate the position of free-field sounds, either with a hand-pointing or with a head-turning response. Pointing movements were performed either blindfolded or with eyes open, but no visual feedback was available about sound position or the motor response. All RBD patients showed some impairment in sound localisation, particularly for sounds towards the contralesional side. In addition, task-irrelevant vision was more detrimental for hand-pointing than head turning responses, only for neglect patients. We propose that this finding reflects visual coding of sound position when the eyes are open, which extends the pathological visuospatial bias of neglect patients to sound localisation. Moreover, the absence of any modulatory effects of ambient vision when head turning responses were adopted suggests task-dependent visual coding of sound position, in agreement with multiple frames of reference for sound localisation. PMID- 12544840 TI - The responses of visual cortical neurons encode differences across saccades. AB - Primate vision consists mostly of periods of stable fixation separated by rapid saccadic eye movements. Each saccade brings a new scene onto the retina, and each new scene results in a burst of activity in the neurons of visual cortex. It might be expected that the activity of these neurons should only represent what is on the retina now, much as a video camera hooked up to a television only displays what the camera is currently pointed at. However, we show here that this is not the case. Recording from 25 primary visual cortical neurons in an awake primate demonstrated that the responses to the saccade-induced presentation of a stimulus within a neuron's receptive field (RF) are typically suppressed by the presence of a stimulus in the RF before the saccade. Flashing stimuli on with the eyes stationary showed, on average, suppressive effects of similar magnitude, suggesting that the mechanism is simple adaptation. However, while the mechanism may be simple, the implications for the operation of the visual system are not. The activity of visual cortical neurons does not represent just the current retinal image, but also the differences between the current retinal image and the previous one. These results suggest that the current approach of studying the visual system, which concentrates on determining the relationship between a single stimulus and a single response, may have to be modified to take into account the timing of retinal image changes that occurs in normal vision. PMID- 12544841 TI - Strain-specific caspase-3-dependent programmed cell death in the early developing mouse forebrain. AB - Caspase-3-deficient 129/Sv mice show hyperplasia of the forebrain at embryonic day (E) 10.5, which suggests that caspase-3-dependent programmed cell death (PCD) plays an essential role in brain morphogenesis prior to neurogenesis. However, little is known about region-specific caspase-3-dependent PCD in the developing forebrain. We examined the PCD region in the early developmental brain at E9.5 by whole mount terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL). In addition to hindbrain, TUNEL-reactivity was detected in the ventral forebrain and in the caudal portion of the front nasal region, just behind the regions expressing fgf-8 and otx-2. It has been shown recently that brain hyperplasia induced by caspase-3-deficiency is mouse strain-dependent; such that brain abnormalities were observed in caspase-3-deficient 129/Sv mice but not in caspase-3-deficient C57BL/6 mice. We examined the caspase-3-dependent PCD in the ventral forebrain of 129/Sv and C57BL/6 mouse embryos (E8.5-9 and E9.5) by double staining of TUNEL and antiserum against the active form of caspase-3 (anti m3D175). TUNEL/anti-m3D175 reactivity in the ventral forebrain was mouse strain dependent, such that many TUNEL/anti-m3D175-positive cells were detected in the ventral forebrains of 129/Sv mice, but were not observed in C57BL/6 mice. Thus, it is likely that this region is the site of the strain-specific caspase-3 dependent PCD. A strain-dependent 'modulator' that regulates both caspase-3 dependent and -independent cell death pathways may control PCD in the ventral forebrain at E8.5-9.5. PMID- 12544842 TI - Human immunoglobulins and Fc fragments promote microtubule assembly via tau proteins and induce conformational changes of neuronal microtubules in vitro. AB - The influence of human immunoglobulins (Ig) in neuronal cytoskeleton stability was studied in vitro. Here we show that human Ig and Fc fragments stimulate animal and human microtubule assembly by binding to microtubules via tau isoforms. In presence of Ig, microtubules show increased aggregation, twisting and rigidity. Non-immune Ig and Fc fragments promote microtubule assembly in temperature-dependent manner and stabilize microtubules at a molecular ratio of 1 Ig per 4 tubulin dimers. These in vitro data provide an experimental support for an immuno-mediated modulation of the cytoskeleton. In conjunction with previous neuropathological data, they suggest that Ig could participate in early stages of neurodegeneration by affecting the microtubule stability in vivo. PMID- 12544843 TI - Early intraneuronal Abeta deposition in the hippocampus of APP transgenic mice. AB - Increasing evidence suggests that intraneuronal amyloid-beta (Abeta) accumulation may be an early event in Alzheimer's disease (AD) pathogenesis. However direct in vivo evidence regarding initial Abeta seeding is missing. Using an APP transgenic mouse model, our sensitive immunocytochemical procedures revealed a novel intraneuronal Abeta deposition in the somas of hippocampal CA1/subiculum neurons far in advance of the occurrence of extracellular Abetaplaques. These deposits increased exponentially with age and were elevated approximately 4-fold (p < 0.001) by high fat/high cholesterol diet. Abeta40 and Abeta42 were the major constituents of these deposits and were co-localized with lysosomal markers. Our results are consistent with the notion that the earliest Abeta deposition occurs intraneuronally, prior to extracellular amyloid plaque formation. PMID- 12544844 TI - Neuronal correlates of familiarity-driven decisions in artificial grammar learning. AB - It has been proposed on the basis of behavioural data that grammaticality judgments in implicit artificial grammar learning paradigms are largely driven by priming based on fragment familiarity. A prediction that follows from this account is that neural deactivation, a common correlate of repetition priming, should be observed for grammatical compared to ungrammatical stimuli. We conducted an event-related fMRI study to investigate neuronal correlates of such fragment-based priming. In a study phase, participants performed a short-term memory task on a series of strings of pseudofont characters. Scanning was performed in a subsequent test phase in which participants classified new strings as either grammatical or ungrammatical. Test strings differed systematically from training strings in terms of exemplar and fragment similarity. Behaviourally, participants classified strings as grammatical based on fragment familiarity. Differential activity was evident during string classification as reduced activity in left lateral occipital complex and bilateral lingual gyri for strings with high fragment familiarity compared to strings with low fragment familiarity. Thus, consistent with the hypothesis, neuronal facilitation in extrastriate occipital regions may constitute one basis of implicit grammaticality decisions based on fragment priming. PMID- 12544845 TI - Cortical reorganization induced by task-oriented training in chronic hemiplegic stroke patients. AB - We investigated the effect of task-oriented training (TT) on the cortical activation pattern in four chronic hemiparetic stroke patients. A TT program, consisting of six tasks, which were designed to improve hemiparetic upper extremity function, was performed for 40 min/day, 4 days/week for 4 weeks. The functional status of the affected hand and fMRI were assessed before and after the TT program. fMRI was performed at 1.5 T in parallel with timed finger flexion extension exercises at a fixed rate. The main cortical activation changes with functional recovery were a decrease in the unaffected and an increase in the affected primary sensorimotor cortex activities. In conclusion, it seems that cortical reorganization was induced by the TT program in chronic hemiparetic stroke patients. PMID- 12544846 TI - Differentiation of human neural stem cells into retinal cells. AB - We have previously reported that transplanted human neural stem cells (HNSCs) display extensive migration and positional incorporation into the aged rat brain, which is associated with an improvement in cognitive function. In the current study, to investigate whether HNSCs are capable of differentiating into retinal cells, we treated HNSCs with human transforming growth factor-beta3 (TGF-beta3) under a serum-free differentiation condition. After 5 days of differentiation in vitro we detected opsin-immunopositive cells in the culture treated with TGF beta3. We also transplanted TGF-beta3-treated HNSCs into the rat vitreous cavity. The donor cells migrated and differentiated into opsin-positive cells in the host retinal cell layer. Here we show for the first time that TGF-beta3-treated HNSCs differentiate into retinal cells. PMID- 12544847 TI - Axotomy differentially regulates voltage-gated calcium currents in mice sensory neurones. AB - Medium sized dorsal root ganglion neurones are involved in tactile sensation and responsible for allodynia following nerve injury. We examined the effects of sciatic nerve injury on the expression of low and high voltage-gated calcium currents in medium sized neurones isolated from lumbar dorsal root ganglia of adult mice. Based on the relative expression of these calcium channel types, three populations of medium sized neurones were identified in controls. Type I, II and III populations were characterised respectively by small, predominant and no low voltage-gated current compared to the high voltage-gated current. Five days after nerve injury, calcium current expression was differentially affected by axotomy in these three subsets of medium neurones. Altogether, these results suggest that calcium channels are heterogeneously distributed among the medium sized neurones. This heterogeneity should provide specificity not only to sensory functions but also to sensory responses following nerve injury. PMID- 12544848 TI - Why are antisaccades slower than prosaccades? A novel finding using a new paradigm. AB - Eye movements away from a new object (antisaccades) are slower than towards it (prosaccades). This finding is assumed to reflect the fact that prosaccades to new objects are made reflexively, and that for antisaccades, reflexive eye movements have to be inhibited and antisaccades are generated volitionally. Experiment 1 investigated the relative contribution of saccade inhibition by comparing the latency difference between pro- and antisaccades obtained in the traditional blocked paradigm and in a new paradigm in which oculomotor inhibition across pro- and antisaccades was matched. When inhibition was placed on the oculomotor system, the latency difference between pro- and antisaccades was significantly reduced. Experiment 2 examined the contribution of volitional saccade programming and execution by requiring both pro- and antisaccades to be programmed volitionally. This manipulation did not decrease further the difference between pro- and antisaccades. It is thus concluded that oculomotor inhibition is the main factor leading to long antisaccade latency. The remaining difference is attributed to the reallocation of covert attention from the target location towards the opposite antisaccade location. PMID- 12544849 TI - Evidence for training-induced crossmodal reorganization of cortical functions in trumpet players. AB - The aim of this study was to compare multimodal information processing in the somatosensory and auditory cortices and related multimodal areas in musicians (trumpet players) and non-musicians. Magnetoencephalographic activity (MEG) was recorded in response to five stimulus conditions from 10 professional trumpet players and nine musically untrained control subjects. Somatosensory and auditory stimuli were presented alone or in combination. Our data suggest that musicians, in general, process multisensory stimuli differently to the control group. When stimulating the lip in professional trumpet players, a multimodal interaction (expressed as difference between the multimodal response and the sum of unimodal responses) in the corresponding somatosensory cortex showed a positive peak at 33 ms, which was not found in the control group. Conversely, the control group shows a significant interaction of opposite polarity around 60-80 ms. We suggest that training-induced reorganization in musicians leads to a qualitatively different way to process multisensory information. It favors an early stage of cortical processing, which is modified by the connections between multimodal and auditory neurons from thalamus to primary somatosensory area. PMID- 12544850 TI - Abeta levels in serum, CSF and brain, and cognitive deficits in APP + PS1 transgenic mice. AB - We compared beta-amyloid peptide (Abeta) levels in the serum, CSF and brain (hippocampus) and correlated these with spatial learning in APP+PS1 transgenic mice. Compared with non-transgenic littermates, male 14-month-old APP + PS1 mice were impaired in spatial learning in the water maze. Among the APP + PS1 mice, only the hippocampal insoluble Abeta42 level correlated with spatial memory (r = 0.44). The levels of insoluble Abeta40 and Abeta42 were highly correlated (r = 0.92), and also correlated with soluble hippocampal Abeta42 (r = 0.64/0.69), which further correlated with the CSF Abeta42 (r = 0.52). None of these parameters correlated with serum Abeta40 levels. These findings support the role of insoluble Abeta42 in memory dysfunction and suggest a model with several pools (insoluble, extracellular soluble, CSF) of Abeta being in partial equilibrium with each other. PMID- 12544851 TI - Benign paroxysmal positional vertigo: some recent advances. PMID- 12544852 TI - Migrainous vertigo: development of a pathogenetic model and structured diagnostic interview. AB - PURPOSE OF REVIEW: Vestibular symptoms occur frequently in patients with migraine. This review refines recently proposed diagnostic criteria for migraine related vestibular symptoms, and develops a pathophysiological model for the interface between migraine and the vestibular system. RECENT FINDINGS: The epidemiological link between migraine and vestibular symptoms and signs suggests shared pathogenetic mechanisms. Links between the vestibular nuclei, the trigeminal system, and thalamocortical processing centers provide the basis for the development of a pathophysiological model of migraine-related vertigo. During the last year, several studies have increased understanding of the relationship between migraine and vestibular symptoms. A study of motion sickness and allodynia in migraine patients supports the importance of central mechanisms of sensitization for migraine-related vestibular symptoms. A study has demonstrated effective treatment of vertigo with migraine therapy. The identification of migrainous vertigo, however, is hampered by a lack of standardized assessment criteria for both clinical and research practices. The application of published criteria for the diagnosis of migrainous vertigo allows the development of a standardized, structured assessment interview. SUMMARY: An understanding of the relationship between migraine and the vestibular system increases knowledge of the pathogenesis of both migraine and vertigo. In addition, studies have identified successful treatment, with standard migraine therapies, of vestibular symptoms in patients with both migraine and vertigo. The use of a standardized assessment tool to identify this unique population of patients will help future studies to test both the pathological model and effective treatment options. PMID- 12544853 TI - An update on falls. AB - PURPOSE OF REVIEW: Falls among elderly persons create immense social problems because of their association with physical decline, serious psychosocial consequences, negative impact on the quality of life, and markedly reduced survival. In addition, falls pose high costs to the public health service. For these reasons, falls remain a popular topic for research. Here, we review several interesting developments that have been published during the past year. RECENT FINDINGS: Considerable attention has been focused on early identification of fallers and prevention strategies for falls or injuries. A remaining challenge is to implement such strategies into daily clinical practice, at acceptable costs and with good acceptance. New evidence has been generated to suggest the importance of high-level processing for postural control, but it remains difficult to predict falls based upon a simple dual tasking test. Neuroimaging studies are now beginning to provide new insights into the role of cortical structures in balance regulation, and how cortical dysfunction might contribute to falls. Posturography studies using multidirectional moving platforms or computer models of balance have provided new insights into falling mechanisms, in particular the role of abnormal protective responses (stepping or grasping) and the negative effects of stiffening. Other studies have shown how a fear of falling, a common sequel to falls, may lead to secondary degeneration of postural control, thus completing a vicious loop. SUMMARY: While these examples illustrate that falls research is very much ongoing, one cannot speak of true progress until the scientific evidence can be implemented into real life, with falls being prevented effectively in individual cases. PMID- 12544854 TI - Neurofibromatosis 2. AB - PURPOSE OF REVIEW: Recent clinical and molecular research on neurofibromatosis 2 (NF2) is reviewed, and the implications for clinical practice and research are discussed. RECENT FINDINGS: NF2 patients who are treated in specialty centers have a significantly lower risk of mortality than those who are treated in non specialty centers. Vestibular schwannoma growth rates in NF2 are generally higher in younger people but are highly variable, even among multiple NF2 patients of similar ages in the same family. Radiation therapy is best reserved for NF2 patients who have particularly aggressive tumors, those who are poor surgical risks, those who refuse surgery, or those who are elderly. In-vivo studies have demonstrated that leptomeningeal cell activation of in mice results in leptomeningeal hyperplasia and meningioma formation. In-vitro studies have identified molecules that interact with the product (merlin or schwannomin), some of which (e.g., CD44 and paxillin) may play critical roles in merlin growth regulation. SUMMARY: NF2 patients should be referred to specialty treatment centers for optimal care. Clinical management of multiple patients in NF2 families cannot be based on the expectation of similar vestibular schwannoma growth rates, even when other clinical aspects of disease severity are similar. The availability of accurate mouse models of human NF2-associated tumors and the identification of molecules involved in merlin growth regulation now provide an opportunity to design targeted treatments for schwannomas and meningiomas. PMID- 12544855 TI - Neuro-ophthalmology of mitochondrial diseases. AB - PURPOSE OF REVIEW: To review recent data on mitochondrial diseases with emphasis on their neuro-ophthalmic manifestations. RECENT FINDINGS: Numerous studies have associated mitochondrial diseases with neuro-ophthalmic manifestations. Although there has been an explosion of studies on the genetics of mitochondrial diseases over the past few years, pathogenesis is only partly understood and therapy remains very limited. Over the past year, new mutations in Leber's hereditary optic neuropathy have been reported, and at least three genes associated with autosomal dominant chronic progressive external ophthalmoplegia have been described. These findings allow a better definition of the specific genetic mutations and gene products as well as pathophysiology of Leber's hereditary optic neuropathy and chronic progressive external ophthalmoplegia. The current development of animal models allows a better understanding of the pathophysiology of human mitochondrial diseases. SUMMARY: The afferent and efferent visual pathways within the central nervous system are frequently involved in mitochondrial diseases. Neuro-ophthalmic signs figure prominently and may be the presenting or even sole manifestation of these disorders. The four most common neuro-ophthalmic abnormalities seen in mitochondrial disorders are bilateral optic neuropathy, ophthalmoplegia with ptosis, pigmentary retinopathy, and retrochiasmal visual loss. PMID- 12544856 TI - Vertebro-basilar syndromes causing oculo-motor disorders. AB - PURPOSE OF REVIEW: All functional classes of eye movements require exquisite coordination between cortical, basal ganglia and brainstem centers involved in ocular motor control. Vertebrobasilar stroke may produce a wide spectum of isolated or combined eye-movement disorders. The intent of this article is to summarize the curent knowledge of eye-movement disorders occurring in infarcts involving the different arterial territories of the vertebrobasilar circulation. RECENT FINDINGS: In the last few years there has been an improvement in our understanding of pathophysiological mechanisms that are related mainly to abnormal vergence disorders due to thalamic-subthalamic infarcts, as well as of the peculiar symptoms resulting from otolith pathway involvement occurring in lateral medullary infarcts. Moreover, progress in neuroimaging technology has implicated neurovascular contact of the trochlear nerve in instances of superior oblique myokimia. SUMMARY: Eye-movement disorders commonly occur in vertebrobasilar stroke, although they are often unappreciated. They may make it possible to establish specific anatomical correlates, as well as the probable nature of the underlying pathology. PMID- 12544857 TI - Visual search and its disorders. AB - PURPOSE OF REVIEW: A greater understanding of the underlying component mechanisms of normal visual search provides explanations for disturbances seen in certain neurological conditions. This review focuses on recent advances in this field which bear on the neurology of visual search in health and disease. RECENT FINDINGS: Foremost, visual search requires a normal apparatus for the application of attentional resources to the visual environment and, with that facility lost in hemispatial neglect, search becomes uselessly stuck within one portion of the field. New evidence suggests that loss of normal registration of where the eyes have been compounds the problem. Even if attention can be deployed flexibly, its parameters must be chosen strategically, in terms of saccade amplitude, size of attentional window at each fixation and search path taken. Evidence is growing that the prefrontal cortex plays a complex role in this strategic control. Rehabilitation strategies of the future may be tailored according to which component functions have been lost in different patient groups. SUMMARY: Visual search is a dominant human activity and provides not only a window into how brain function is deranged after structural damage, but also offers the prospect of an ideal modality through which to deliver future behavioural therapies. New techniques have advanced our understanding of the physiology of visual search enormously in the past few decades. The time is now ripe in which to begin to integrate these findings into our understanding of the pathophysiology and treatment prospects of neurological disorders like hemispatial neglect, hemianopia and other deficits after stroke. PMID- 12544858 TI - Imaging of acute ischemic brain injury: the return of computed tomography. AB - PURPOSE OF REVIEW: Classical and modern computed tomography imaging techniques, including conventional computed tomography, perfusion-computed tomography and computed tomography-angiography for acute stroke are reviewed. RECENT FINDINGS: Thrombolysis has become an approved therapy for acute stroke. However, many stroke patients do not benefit from such treatment, since the presently used criteria are very restrictive, notably with respect to the accepted time-window. Even so, a significant rate of intracranial hemorrhage still occurs. Conventional cerebral computed tomography has been proposed as a selection tool for acute stroke patients. Recently, more modern computed tomography techniques, referred to as functional computed tomography or perfusion-computed tomography, have been introduced. Such perfusion-computed tomography techniques are competing favorably with diffusion-weighted/perfusion-weighted magnetic resonance imaging in the delineation of the penumbra. They are more easy to perform, because they are readily available and accessible in emergency settings, and less time-consuming. Furthermore, perfusion-computed tomography combined with computed tomography angiography affords a comprehensive non-invasive survey of acute stroke patients as early as at the time of their emergency admission, with accurate demonstration of the site of arterial occlusion and its hemodynamic and pathophysiological repercussions for the brain parenchyma. SUMMARY: A renewal of computed tomography in the field of stroke has been afforded by the introduction of functional techniques such as perfusion-computed tomography, which provides promising insights into cerebral tissue viability and can be used as a guidance tool for therapy when combined with computed tomography-angiography. PMID- 12544859 TI - Lessons in experimental ischemia for clinical stroke medicine. AB - PURPOSE OF REVIEW: This review covers experimental developments in the laboratory and their translation to clinical stroke trials over the year from 3 August 2001 to 2002. RECENT FINDINGS: Recent findings include novel observations in the areas of excitotoxicity, free radical injury, neuro-inflammation and apoptosis. A key clinical finding this year has been the translation of the effects of hypothermia in global ischemia to two successful trials in cardiac arrest with patients cooled within 4-8 h following resuscitation achieving good neurological outcomes, as compared with their normothermic controls. An era of molecular imaging in stroke research is presaged by the first reports of enhanced magnetic resonance or labeling with supramagnetic contrast agents. SUMMARY: Although none of the drugs in focal ischemia has translated from experimental models, for the first time there is evidence of cytoprotection for the brain that has been translated from the laboratory to man. PMID- 12544860 TI - Thrombolysis for acute stroke. AB - PURPOSE OF REVIEW: Stroke thrombolysis is in the process of moving from academic centers into community-based standard therapy in experienced centers. Recently published data on patient selection, imaging and other technical or organizational advances are reviewed in order to improve the safety and efficacy of stroke thrombolysis and identify areas in which more study is needed. RECENT FINDINGS: During the past year, several open series were published on the application of thrombolysis in community-based settings, which have mostly reproduced the outcome described in randomized, placebo-controlled recombinant tissue plasminogen activator cohorts. New information has been produced to identify the clinical parameters that are associated with good and adverse outcomes after stroke thrombolysis. Elevated blood glucose decreases the likelihood of a good outcome and increases that of cerebral hemorrhagic change, but the effect of early glucose reduction needs to be investigated. Non-contrast computed tomography still prevails as the standard imaging method in patient selection, but computed tomography angiography can be added within a reasonable time to furnish vascular diagnosis. Perfusion-weighted imaging is used to quantify the fraction of brain tissue salvaged by intravenous thrombolysis, and may be used in the future to select patients with still viable penumbral brain tissue, even beyond a 3-h time-window, if efficacy can be proved. SUMMARY: Thrombolysis with recombinant tissue plasminogen activator is well tolerated and effective within 3 h of symptoms, and new trials will determine if extension of this time-window or new target populations for thrombolysis, such as children with stroke, can be substantiated. An unfinished task remains in the education of health personnel; imperatives stemming from pathophysiology need to penetrate attitudes on acute stroke at all levels before acute therapies such as thrombolysis can have an impact on stroke outcome in general. PMID- 12544861 TI - The role of blood pressure lowering before and after stroke. AB - PURPOSE OF REVIEW: Elevated blood pressure is one of the most potent risk factors for first ever and recurrent stroke as well as influencing early outcome after acute stroke. There have been a number of significant randomized controlled trials which may influence management in each of these three categories. RECENT FINDINGS: For primary prevention, the recent information from the Heart Outcomes Prevention Evaluation, Losartan Intervention for Endpoint Reduction to Hypertension, Study on Cognition and Prognosis in the Elderly and Australian National Blood Pressure Study support the view that blood pressure lowering protects against stroke regardless of baseline blood pressure level. There is some evidence that blockade of the angiotensin system may give additional protection. For secondary prevention, evidence from the Perindopril Protection against Recurrent Stroke Study shows that blood pressure lowering with perindopril based therapy reduces fatal or non-fatal stroke events, again in hypertensive or normotensive individuals. There is uncertainty about blood pressure lowering in acute stroke, although presentation of the recent Acute Candesartan Cilexetil Evaluation in Stroke Survivors trial in which there was significant protection against vascular events using candesartan suggests that further studies should be undertaken. SUMMARY: Blood pressure lowering for primary prevention of stroke should be undertaken using a variety of therapeutic agents. For secondary stroke prevention perindopril based therapy should be used based on current evidence. Uncertainty still exists as to whether blood pressure lowering in the acute stroke setting is safe or improves outcomes. PMID- 12544862 TI - Sleep-disordered breathing and stroke. AB - PURPOSE OF REVIEW: A growing number of studies suggest that there is a close relationship between sleep-disordered breathing (SDB) and stroke. Since the prevalence of SDB is high among acute stroke patients and since oxygen desaturations are potentially dangerous, SDB may influence not only stroke risk, but also stroke outcome and recurrence. This review summarizes recent data on this issue, pointing out existing evidence but also defining issues that still need to be clarified. RECENT FINDINGS: Recent studies further supported the hypothesis of a link between SDB and stroke: (1) although stroke occurs most frequently in the morning hours, i.e. shortly after awakening, stroke events take place predominantly during sleep in SDB patients; (2) a large cross-sectional study confirmed the association of SDB not only with coronary heart disease but also with stroke; (3) several works suggest that SDB may increase stroke risk through various mechanisms including blood pressure swings, endothelial dysfunctions, prothrombotic coagulation shifts, pro-inflammatory states, increased platelet aggregation and atherosclerosis; (4) one study in acute stroke patients demonstrated that SDB is associated with early neurological worsening although not with final stroke outcome; (5) treatment trials with continuous positive airway pressure (CPAP) have demonstrated mild reductions of diastolic and systolic blood pressure in SDB patients, an effect that is expected, however unproven, to decrease stroke risk. SUMMARY: Despite considerable progress regarding our understanding about the role of SDB in vascular disease, further studies are needed to determine whether cerebrovascular morbidity is related to SDB independent of associated vascular risk factors, whether SDB has indeed a negative impact on stroke progression and whether CPAP treatment ameliorates stroke outcome. PMID- 12544863 TI - Expression of A20 in the vessel wall of rat-kidney allografts correlates with protection from transplant arteriosclerosis. AB - BACKGROUND: Chronic rejection with development of transplant arteriosclerosis is the major culprit involved in loss of kidney allografts. The allografts' fate was thought to depend on the intensity of the host immune responses and the potency of immunosuppressive regimens. Recent data suggests that grafts contribute to their own survival by way of up-regulation of "cytoprotective" genes. METHODS: We analyzed the expression of four cytoprotective genes, A20, Bcl-2, Bcl-x(L) and heme oxygenase (HO)-1, in three rat renal allograft models of chronic rejection: Fisher 344-Lewis (F344/Lew), Dark Agouti-Brown Norway (DA/BN), and DA-Wistar Furth (WF). We chose these genes for their known anti-inflammatory and anti apoptotic function in endothelial cells (EC) and the atheroprotective function of A20 in smooth muscle cells (SMC). RESULTS: Twenty-eight and 9 weeks following transplantation, F344/Lew and DA/BN transplants had stable graft function. Histopathologic analysis showed moderate tissue damage, minimal cellular infiltrates, and preserved vascular integrity correlating with high expression of A20 in SMC. Conversely, impaired allograft function in the DA/WF combination with substantial transplant arteriosclerosis was noted in 60% of the grafts correlating with absent or decreased A20 expression in EC and SMC. In all combinations, expression of HO-1, Bcl-2, and Bcl-x(L) colocalized with infiltrating cells and was not informative on the graft status. CONCLUSIONS: We demonstrate for the first time a strict correlation between A20 expression in the vessel and the absence of transplant arteriosclerosis in rat kidney-allograft models. This data is similar to data obtained in human kidney allografts and suggests that A20 may represent a novel therapeutic target for the prevention of chronic allograft rejection. PMID- 12544864 TI - Improvement in human decay accelerating factor transgenic porcine kidney xenograft rejection with intravenous administration of gas914, a polymeric form of alphaGAL. AB - BACKGROUND: The present study was undertaken to determine whether intravenous administration of GAS914, a polymeric form of alphaGal, would minimize porcine kidney xenograft rejection in baboons. Human decay accelerating factor renal xenografts were transplanted into 16 baboon recipients. METHODS: Baseline immunosuppression for all groups included cyclosporine A, cyclophosphamide, SDZ RAD, and methylprednisolone. Group 1 received only baseline immunosuppression; group 2 animals received low-dose GAS914 with baseline immunosuppression; group 3 animals received high dose GAS914 with high-dose baseline immunosuppression; and animals from group 4 received high-dose GAS914 and low-dose baseline immunosuppression. RESULTS: None of the animals in this study developed hyperacute rejection. Intravenous administration of GAS914 significantly reduced xenoreactive antibodies as measured by antiporcine hemolytic assays and anti-Gal (immunoglobulin [Ig] G and IgM) antibody assays. Rejection was less severe in the GAS914-treated group. Only 25% (3 of 12) of GAS914-treated animals were killed as a result of rejection, whereas 75% (three of four) of non-GAS914-treated animals were killed because of terminal rejection (P<0.01). Protocol biopsies demonstrated that the degree of acute humoral xenograft rejection (AHXR) was reduced in the GAS914-treated animals compared with non-GAS914-treated animals. CONCLUSION: The intravenous administration of GAS914 reduces xenoreactive antibody levels and reduces the degree of porcine kidney xenograft rejection, but does not improve survival. AHXR and drug toxicity remain major barriers to the long-term success of xenotransplantation. PMID- 12544865 TI - Preconditioning with cyclosporine A or FK506 differentially regulates mitogen activated protein kinase expression in rat kidneys with ischemia/reperfusion injury. AB - BACKGROUND: The signaling pathways of mitogen-activated protein kinases (MAPKs) are important molecular components responsible for ischemia/reperfusion (I/R) injury in the kidneys. Preconditioning with cyclosporine A (CsA) or FK506 reduces subsequent I/R injury. We studied the effect of preconditioning with CsA or FK506 on MAPK expression in ischemic rat kidneys. METHODS: Two separate studies were performed using Sprague-Dawley rats. First, MAPK (extracellular signal-regulated kinase [ERK], jun N-terminal kinase [JNK], p38) expressions were observed at 0, 10, 20, 30, 60, 120, and 1,440 min after I/R injury. Second, the effects of preconditioning with CsA or FK506 on MAPK expressions were observed in rat kidneys with I/R injury. I/R injury was induced by clamping both renal arteries for 45 min. Rats were pretreated with intravenous (IV) CsA (3 mg/kg) or IV FK506 (0.3 mg/kg) 6 hr before I/R injury and killed 30 min later. Expression of MAPK was measured using immunoblot and immunohistochemistry. RESULTS: MAPK (ERK, JNK, p38) expressions were significantly increased in kidneys with I/R injury compared with sham-operated controls, and immunohistochemistry revealed increased MAPK immunoreactivity in renal tubules of the outer medulla. Kidneys preconditioned with low-dose CsA or FK506 showed significantly increased ERK expression compared with kidneys with I/R injury alone (CsA, 9.5- vs. 4.5-fold; FK506 10.4- vs. 4.5 fold: P<0.05) but showed decreased JNK (CsA, 3.8- vs. 5.3-fold; FK506, 3.4- vs. 5.3-fold: P<0.05) and p38 expression (CsA, 2.5- vs. 3.7-fold; FK506, 2.1- vs. 3.7 fold: P<0.05). CONCLUSIONS: Preconditioning with CsA or FK506 differentially regulates the expression of MAPK in rat kidneys with I/R injury, and this may explain the remarkable protective effects of these agents. PMID- 12544866 TI - Split tolerance to a composite tissue allograft in a swine model. AB - BACKGROUND: The antigenicity of skin is a major obstacle to expanding human composite tissue transplantation. For example, multiple rejection episodes of the skin have been noted in clinical hand transplant patients. We have previously demonstrated tolerance to vascularized musculoskeletal allografts in major histocompatibility complex (MHC)-matched miniature swine treated with 12 days of cyclosporine. This regimen did not reproducibly lead to tolerance to subsequent frozen donor skin grafts. However, such skin grafts did not have a primary vascular supply. The aim of this study was to determine if tolerance to limb allografts with a vascularized skin component could be achieved with MHC matching and a 12-day course of immunosuppression. METHODS: Hind limb grafts harvested with a 100 cm(2) cutaneous paddle were transplanted heterotopically into six MHC matched, minor antigen-mismatched miniature swine. All animals received a 12-day course of cyclosporine. One control animal was not immunosuppressed. Grafts were evaluated with biweekly biopsies and tissue viability determined by histologic analysis. To test for sensitization, frozen donor skin grafts were applied to all animals that survived to postoperative day 100. RESULTS: All treated animals (n=6) were tolerant to their musculoskeletal allografts at the time of necropsy (>100 days) regardless of the status of the epidermis. One animal demonstrated tolerance to the skin for more than 180 days. The other five animals demonstrated prolonged survival of the epidermal portion of the graft. The control animal rejected the graft epidermis at 10 days postoperatively. Frozen donor skin grafts demonstrated accelerated rejection (<10 days) in three of the animals and led to simultaneous rejection of both the epidermis of the allograft and the skin graft in the long-term tolerant animal. The rejection of the skin grafts did not break tolerance to the musculoskeletal portion in any of the animals. CONCLUSIONS: All animals exhibited indefinite survival of the musculoskeletal portion of their allografts but only prolonged survival of the epidermis. The loss of the graft skin appears to be the result of an isolated immune reaction to the skin, and, in particular, the epidermis. This observation is further substantiated by the accelerated rejection of secondarily placed frozen donor skin grafts. PMID- 12544867 TI - Induction of tolerance to hind limb allografts in rats receiving cyclosporine A and antilymphocyte serum: effect of duration of the treatment. AB - BACKGROUND: This study assessed the ability of antilymphocyte serum (ALS) and cyclosporine A (CsA) to induce tolerance for hind limb composite tissue allograft in rats without chronic immunosuppression. METHODS: Hind limb transplantations were performed in Lewis-Brown-Norway (LBN, RT1(1+n)) and Lewis (LEW, RT1(1)) rats. Treatment consisted of ALS only (0.4 mL/kg), CsA only (16 mg/kg), and a combination of CsA and ALS, and it was administered 12 hr before surgery at three different intervals (7, 14, and 21 days). Long-term survivors were tested for tolerance by standard skin grafting from the recipient (LEW), the donor (LBN), and the third party (ACI, RT1 ) 60 days after cessation of the treatment and by mixed lymphocyte reaction at 100 days. T-cell lines were analyzed with flow cytometry. RESULTS: Single use of ALS in all treatment intervals did not prolong allograft survival. Single use of CsA extended survival up to 23 days in the 21 day protocol group. CsA and ALS caused indefinite survival in two of six rats in the 14-day protocol and in all six rats in the 21-day protocol (>420 days). The six long-term survivors in the 21-day protocol accepted the skin grafts from the donor (LBN) and the recipient (LEW) and rejected third-party grafts (ACI). Tolerant animals showed a donor-specific hematopoietic chimerism of 35% to 42% in the peripheral blood. Mixed lymphocyte reaction assay demonstrated tolerance to the host and donor alloantigens and increased response to the third party. CONCLUSIONS: Administration of CsA and ALS for 21 days induced donor-specific tolerance in the recipients of the rat hind limb composite tissue allografts. The mechanism of tolerance should be investigated further. PMID- 12544868 TI - Randomized double-blind study of immunoprophylaxis with basiliximab, a chimeric anti-interleukin-2 receptor monoclonal antibody, in combination with mycophenolate mofetil-containing triple therapy in renal transplantation. AB - BACKGROUND: Acute rejection remains a major problem in renal transplantation. Immunoprophylaxis with basiliximab (Simulect) has achieved significant reductions in acute rejection episodes in renal allograft recipients receiving dual immunosuppression. This study explored the tolerability and cumulative benefit of combining basiliximab with triple-drug therapy-cyclosporine (USP Modified, Neoral), mycophenolate mofetil, and steroids. METHODS: In a randomized, double blind, placebo-controlled, multicenter study, 123 kidney transplant recipients received either basiliximab at 20 mg before transplantation (day 0) and 20 mg on day 4 (n=59), or placebo (n=64). All received triple-drug immunosuppression and were followed for 6 months. RESULTS: Tolerability of basiliximab was equivalent to placebo, with no increase in serious adverse events, infection, malignancy, or posttransplant lymphoproliferative disorder. At 6 months, there were trends in favor of basiliximab over placebo in the incidences of first biopsy-confirmed acute rejection (15.3% vs. 26.6%, P=NS) and of acute rejection treated with antibody (5.1% vs. 15.6%, P=NS). Kaplan-Meier estimates at 4 weeks and 6 months were significantly in favor of basiliximab treatment for first acute rejection, biopsy-confirmed rejection, rejection episodes treated with antibody therapy, and treatment failure. Renal function improved more rapidly in the basiliximab group, with mean creatinine clearance at week 2 being 54.7 mL/min versus 43.2 mL/min for placebo (P=0.034). At 12 months, patient survival was 100% in both groups; graft survival was 94.9% with basiliximab and 92.2% with placebo. CONCLUSIONS: Basiliximab immunoprophylaxis is safe, well tolerated, and shows a trend toward reduction in number of acute rejection episodes in renal transplant patients receiving cyclosporine, mycophenolate mofetil, and steroids. PMID- 12544869 TI - Single human leukocyte antigen flow cytometry beads for accurate identification of human leukocyte antigen antibody specificities. AB - BACKGROUND: It is difficult to assign antibody specificity for highly sensitized patients using a cell panel with multiple antigens per reaction. We describe here a single antigen bead panel for accurate identification of human leukocyte antigen (HLA) antibody specificities by flow cytometry. METHODS: A total of 110 single recombinant HLAs, including 34 A locus alleles, 57 B locus alleles, and 19 C locus alleles, were produced by a mammalian expression system. These single antigens were coated onto eight different colored microbeads, which were mixed together in one tube for simultaneous detection of HLA antibodies against eight different antigens per flow cytometry test. RESULTS: Single HLA reacted specifically with the serologically defined monoclonal antibodies. The single antigen panel provided higher resolution than the regular cell panel for antibody detection by uncovering the masked specificities. Single antigens also provided higher sensitivity than the multiple antigens coated onto beads for HLA antibody detection as demonstrated by serum dilution studies. In 10 sera from patients who had rejected a kidney transplant, single antigen beads identified antibodies to 31 of 35 antigens that were mismatched in the donor. Most important, none of the reactions were against antigens present in the recipient. CONCLUSION: An accurate and sensitive HLA antibody detection method is described using flow cytometry beads coated with single HLAs produced by recombinant technology. The single antigen beads should be useful in predicting negative crossmatch in highly sensitized organ recipients and highly sensitized patients requiring platelets. PMID- 12544870 TI - Bone mineral density changes within six months of renal transplantation. AB - BACKGROUND: The effective use of new steroid-sparing immunosuppressive regimens may lower cumulative glucocorticoid use among renal transplant recipients. However, it is unknown what effect this therapeutic trend has had on bone disease. METHODS: Unselected newly transplanted inpatients (n=45) were identified and comprehensively evaluated for metabolic bone disease at a median of 16 days (range 9-33) posttransplant. A follow-up evaluation was conducted a median of 5.7 months (range 4.8-9.3) later. Follow-up values for bone mineral density (BMD) and select laboratories were compared with baseline values using nonparametric statistics. Odds ratios (ORs) and 95% confidence intervals (CIs) were used to describe the associations of baseline characteristics, select laboratory values, and cumulative prednisone and cyclosporine use with spinal BMD loss and were calculated using logistic regression. RESULTS: A significant decrease in intact parathyroid hormone (P<0.001) and a significant increase in calcitriol (P=0.02) were noted postengraftment. At follow-up, subjects had lost a mean of 2.4% BMD at the lumbar spine (P=0.003) but did not experience significant declines at the femoral neck. The highest tertiles of cumulative prednisone (OR=28.4; 95% CI 2.5 329 and OR=15.8; 95% CI 1.4-179, respectively) and past alcohol use (OR=9.3; 95% CI 1.46-58.5) were significantly associated with spinal BMD loss. CONCLUSIONS: Significant loss in lumbar BMD occurred within 6 months of transplantation in more than one third of a prospective cohort of renal transplant recipients. Lumbar bone loss seemed to be mediated primarily by glucocorticoid dose and a history of alcohol use. PMID- 12544871 TI - Combination therapy of mycophenolate mofetil and rapamycin in prevention of chronic renal allograft rejection in the rat. AB - BACKGROUND: Chronic rejection is the leading cause of long-term allograft loss. Until now, no therapy has been recognized as being efficient in its prevention. In addition to their immunosuppressive activity, mycophenolate mofetil (MMF) and rapamycin (RAPA) show diverse properties against vascular smooth muscle cell activity, cell-adhesion molecule expression, and ischemia-reperfusion injury. The combination effect of MMF and RAPA was tested to prevent chronic renal allograft rejection in the rat in this study. METHODS: Nephrectomized Lewis recipients underwent orthotopic transplantation with Fisher (F344) kidneys (allograft groups) or Lewis kidneys (isograft control). The initial episode of acute rejection was controlled with a short course of cyclosporine A (CsA) (1.5 mg/kg/day for 10 days). From weeks 4 to 20, animals were thereafter treated every other day either with vehicle, MMF (20 mg/kg), RAPA (0.8 mg/kg), or MMF (20 mg/kg) plus RAPA (0.8 mg/kg) in combination. Animals were sequentially killed at serial intervals over a follow-up of 50 weeks, and histologic study was performed on harvested kidneys according to the Banff working classification for allograft pathology. RESULTS: Animals treated with MMF or RAPA alone showed a Banff sum score similar to the allograft control group (6.31+/-1.01 and 7.27+/-1.14 vs. 7.21+/-1.14, respectively; P>0.05). When the recipient rats were treated with MMF and RAPA in combination, it resulted in a clinically and statistically significant reduction of Banff sum score (4.21+/-0.79, P<0.01), with specific inhibition of vascular fibrous intimal thickening, allograft glomerulopathy, and interstitial fibrosis. CONCLUSION: Over a 50-week study, concomitant therapy of MMF and RAPA prevents chronic renal allograft rejection, probably through reduction of ischemic and cytotoxic degenerative changes. These results warrant further investigation in the combination of MMF and RAPA as anti-chronic rejection therapy in clinical transplantation. PMID- 12544872 TI - Prophylactic deoxyspergualin treatment in living-related renal-transplant recipients transfused with donor-specific blood. AB - BACKGROUND: Deoxyspergualin (DSG) prophylaxis has improved long-term graft survival in living-related renal-transplant recipients transfused with donor specific blood (DST). We examined the influence of acute rejection (AR) on graft survival in these patients. METHODS: The study groups consisted of either historic control recipients without DSG (group A, n=64, 1985-1989) and recipients with DSG as the initial immunosuppressive agent (group B, n=76, 1989-1995). Both groups received DST from a one-haplotype identical donor and were treated with cyclosporine-based immunosuppression. Rejection was classified into accelerated rejection (Acc, within 5 days), AR (from 6 days-3 months), and late AR (LAR, from 4 months-1 year). RESULTS: Overall 5-year graft survival rates were significantly higher in group B than group A (89.5 vs. 73.4%, P=0.0070). Each group was then subdivided on the basis of whether or not they had an episode of Acc, AR, or LAR. In group A, 5-year graft survival rate was not affected the presence or absence of Acc (75.0 vs. 73.1%), and it was influenced significantly by the presence or absence of AR (50.0 vs. 85.7%, P=0.0012) or LAR (46.7 vs. 81.6%, P<0.0001). In group B, 5-year graft survival did not change significantly by the presence or absence of Acc (100 vs. 88.7%), AR (81.8 vs. 92.6%), or LAR (81.0 vs. 92.7%). CONCLUSIONS: Prophylactic use of DSG in living-related renal-transplant recipients treated with DST improves long-term graft survival, even in patients with AR episodes. PMID- 12544873 TI - Recurrent and de novo diabetic nephropathy in renal allografts. AB - BACKGROUND: Histologic findings of diabetic nephropathy (DN) are observed in allografts of patients with pretransplant (PreTx) diabetes mellitus (DM) and in patients who develop DM posttransplant (PostTx). Patients with allograft biopsies (Bx) were retrospectively studied to determine the incidence of recurrent and de novo DN and to ascertain what, if any, risk factors predispose to histologic DN in either patient population. METHODS: From the renal transplant services at four hospitals from 1992 to 2000, the authors identified all patients with PreTxDM and PostTxDM (n=81). Those with renal biopsies performed >/=18 months PostTx were classified according to the presence or absence of histologic DN (Bx-positive, n=23; Bx-negative, n=35). Patients were then subdivided into four categories recurrent DN (n=16), de novo DN (n=7), no recurrent DN (n=27), and no de novo DN (n=8)-for analyses. RESULTS: Among these 58 patients, 74.1% had PreTx and 25.9% had PostTx diabetes. Of those with histologic DN, 69.6% were recurrent DN and 30.4% were de novo DN, making de novo DN at least as likely to develop as recurrent DN. After the onset of diabetes in the de novo population, the time to development of histologic DN was similar in the recurrent and the de novo patients (6.68+/-3.86 years vs. 5.90+/-3.13 years, P=0.66) and more rapid than previously reported. Apart from a more frequent family history of hypertension in patients with allograft DN compared with those without allograft DN, known risk factors for the development of native DN did not significantly differ among patients in the four cohorts. Proposed risk factors related to transplantation did not correlate with the development of recurrent or de novo DN. CONCLUSION: Among patients with histologic DN, de novo DN occurred at least as frequently as recurrent DN, and the time to onset of histologically apparent DN was more rapid than previously reported. Neither the usual clinical predictors of DN nor clinical variables related to transplantation clearly distinguished the group with DN from the group without it, potentially implicating novel mechanisms in its pathogenesis. PMID- 12544874 TI - Chemokine and chemokine receptor gene expression indicates acute rejection of human cardiac transplants. AB - BACKGROUND: Factors directing T-cell infiltration into allografts during acute rejection remain poorly defined. Chemokines have been shown to mediate leukocyte recruitment into allografts in animal models of rejection. The goal of this study was to test the presence and levels of chemokine and receptor gene expression in serial endomyocardial biopsy specimens from heart transplant patients and to correlate the levels observed with histopathologic rejection grade. METHODS: Three hundred sixteen serial endomyocardial biopsy specimens from 30 heart transplant patients were obtained during the clinically scheduled surveillance heart biopsy program. The follow-up period was 1 year. The expression of interferon (IFN)-gamma inducible protein (IP)-10, monokine induced by IFN-gamma (Mig), interferon-inducible T-cell alpha chemoattractant (I-TAC), regulated on activation normal T-cell expressed and secreted (RANTES), monocyte chemotactic protein (MCP)-1, interleukin (IL)-8, and the receptors CXCR3 and CCR5 were tested using quantitative, real-time polymerase chain reaction. Biopsy samples were examined histologically to assign rejection grade. RESULTS: Expression of IP-10, Mig, I-TAC, RANTES, CXCR3, and CCR5, but not MCP-1 and IL-8, increased significantly in both grade 2 and grade 3 rejection (P100,000 copies/microL). BKV DNA was also detected in the plasma of three patients. All the patients were treated with intravenous low-dose cidofovir (0.25 1 mg/kg per dose, every 2-3 weeks, without probenecid). BK viruria resolved within 4 to 12 weeks (after 1-4 doses) of the cidofovir therapy, and all patients remain with stable renal function 6 to 26 months posttherapy. CONCLUSIONS: Quantitative PCR for BKV is a sensitive and reliable method for following the course of the infection in renal transplant patients. In addition, cidofovir therapy may be useful in the treatment of some of these patients, and its role needs to be investigated further. PMID- 12544882 TI - Death in the first year after kidney transplantation: implications for patients on the transplant waiting list. AB - BACKGROUND: As the kidney transplant waiting list continues to expand, maintaining the medical fitness of transplant candidates will become increasingly difficult. METHODS: To identify patients who are at greatest risk during the wait list period, we performed a Cox regression analysis to determine risk factors for mortality in the first posttransplantation year among 23,546 adult first kidney transplant recipients recorded in the United States Renal Data System between January 1995 and September 1997. RESULTS: In this study population, 4.6% of the patients died in the first posttransplantation year, and cardiac causes were the leading cause (27%) of death. Patients with diabetes (hazard ratio [HR]=1.58; 95% confidence interval [CI], 1.39-1.80), peripheral vascular disease (HR=1.41; 95% CI, 1.11-1.80), or angina (HR=1.38; 95% CI, 1.15-1.65), and patients with a longer duration of end-stage renal disease (HR=1.06 per year; 95% CI, 1.04-1.09) had a higher risk for mortality. Additionally, patients with early acute rejection (HR=1.47; 95% CI, 1.23-1.76), delayed graft function (HR=1.46; 95% CI, 1.25-1.71), and a lower glomerular filtration rate after transplantation were also at increased risk for death within the first posttransplantation year. CONCLUSIONS: Patients with comorbid disease, patients with a long duration of end stage renal disease, and potential recipients of organs at high risk for graft dysfunction should be carefully screened for medical complications before transplantation to achieve the most favorable outcomes. Alternate organ allocation strategies that facilitate patient assessment close to the time of transplantation or that prioritize high-risk patients may also improve outcomes. PMID- 12544884 TI - Improved results after heart-lung transplantation: a 17-year experience. AB - OBJECTIVE: In selected patients with severe end-stage combined cardiopulmonary diseases, heart-lung transplantation (HLTx) remains the only therapeutical option for improving survival and quality of life. PATIENTS AND METHODS: Since 1983, 51 HLTx were done at our institution. Mean patient age was 27+/-12 years with a mean donor age of 25+/-11 years. Indications for HLTx were primary pulmonary hypertension (PPH) in 49% of patients, congenital heart disease in 39%, cystic fibrosis in 6%, and repeat-HLTx in 6%. Eleven patients were younger than 14 years. Among these pediatric patients, the indications were PPH in 55% of patients, pulmonary atresia with severe pulmonary artery hypoplasia in 27%, and cystic fibrosis and cardiomyopathy with fixed pulmonary hypertension in 9% of patients each. Two patients had additional liver transplantation because of chronic aggressive virus hepatitis. For organ preservation, Euro-Collins solution (lung perfusion) and cardioplegic solution according to Bretschneider (heart perfusion) were used until 1994. The University of Wisconsin solution replaced Bretschneider's solution in 1994. Since 1996, Perfadex, a low-potassium dextran based preservation solution, replaced Euro-Collins. All transplantations were done through a median sternotomy until 1994. Thereafter, a transverse thoracotomy was used in patients with suspected adhesions. Until 1995, cyclosporine A, azathioprine, and prednisolone were used for immunosuppression. Since then, tacrolimus replaced cyclosporine A. RESULTS: From 1983 until 1993, perioperative mortality was 35% (6/19). From 1994 on perioperative mortality decreased to 12.5% (4/32). Early mortality was caused by graft failure (n=5), severe bleeding (n=2), multi-organ failure (n=2), and acute rejection (n=1). Cumulative survival rates were 81% after 30 days, 63% after 1 year, and 54% after 5 years, respectively. Since 1994, cumulative survival rates were markedly improved to 87% after 30 days, 81% after 6 months, and 78% after 1 year. There was no death during the first postoperative year among the 11 pediatric patients. Late death was mainly caused by obliterative bronchiolitis (OB; 76%); two patients died because of multi-organ failure or septic complications, respectively, and one patient died within the first postoperative year because of aspergillosis. CONCLUSION: Changes in organ preservation management, surgical techniques, and immunosuppressive therapy significantly improved the short- and mid-term results after HLTx. Long term results can only be improved in cases of successful prevention and treatment of OB. PMID- 12544883 TI - Liver transplant-associated graft-versus-host disease. AB - BACKGROUND: Graft-versus-host disease (GVHD) is an important, underdiagnosed cause of mortality associated with liver transplantation. We identified 12 cases of GVHD among 1,082 liver transplantations performed in patients at our institution between 1991 and 1998. Patients typically developed fever, skin rash, diarrhea, or pancytopenia within 2 to 6 weeks after their transplant. Treatment generally involved increased immune suppression and hematopoietic cytokines (granulocyte colony stimulating factor, granulocyte monocyte colony stimulating factor); however, all but one patient died, most often from sepsis. Early in its course, GVHD was difficult to distinguish from cytomegalovirus disease or drug reactions. The diagnosis was confirmed by demonstration of substantial donor lymphoid chimerism. METHODS: To identify risk factors for severe GVHD, a retrospective analysis was performed comparing index cases with the rest of the cases in our institutional experience. RESULTS: Closely matched human leukocyte antigen recipients, those older than 65 years, and recipients with donors more than 40 years younger were at higher risk for GVHD. One case occurred in a patient with a congenital immunodeficiency. CONCLUSIONS: Liver transplant associated GVHD is a progressive and fatal disease. Future approaches should focus on prevention and might include avoidance of closely matched human leukocyte antigen donors, treatment of the donor to reduce the number of lymphocytes, or reduction of immunosuppression in the early posttransplant period. PMID- 12544885 TI - Impact of human leukocyte antigen matching in liver transplantation. AB - BACKGROUND: Human leukocyte antigen (HLA) compatibilities are beneficial in the setting of kidney transplantation but have demonstrated inconclusive results after liver transplantation. On the basis of recent controversial reports, the authors analyzed the impact of HLA matching in their patients after liver transplantation under modern immunosuppressive drug regimens and new HLA typing techniques with respect to outcome and adverse immunologic events. METHODS: Data from 924 transplants with complete donor-recipient HLA typing were retrospectively analyzed. Immunosuppression was commenced as either cyclosporine A- or tacrolimus-based therapy in different protocols. The follow-up period ranged from 1 to 144.8 months (median, 66 months). RESULTS: The actuarial graft survival was 88% after 1 year and 78.7% after 5 years and was similar in tacrolimus- and cyclosporine A-treated patients. However, cyclosporine A-treated patients underwent significantly more rejection episodes. The number of HLA compatibilities had no influence on graft survival, whereas the number of acute rejections was significantly less in transplants with more HLA compatibilities (P<0.05). Graft survival tended to be improved in patients with chronic hepatitis B and more HLA compatibilities (P=0.05). In contrast, graft survival in transplants for primary sclerosing cholangitis was significantly impaired in the presence of one or two HLA-DR compatibilities (P<0.05). In addition, in autoimmune hepatitis, survival tended to be lower in the presence of more HLA compatibilities (P=0.1). Overall graft survival or frequency of adverse immunologic events was not influenced by any specific donor-recipient HLA allele. CONCLUSION: This study demonstrated fewer acute rejections in transplants with more HLA compatibilities. However, in liver transplantation, a more specific investigation of HLA typing may be necessary, because in some indications HLA antigens play a role in the nature of the disease. Therefore, recurrence of autoimmune disease may be more severe in patients sharing HLA antigens. PMID- 12544886 TI - Rapamycin impairs antigen uptake of human dendritic cells. AB - BACKGROUND: Rapamycin is a recently introduced immunosuppressive agent. Its effect on lymphocytes has been extensively studied. Whether it can also modulate dendritic cell (DC) function is unknown. METHODS: The effect of rapamycin on differentiation, antigen uptake, and the immunostimulatory capacity of human DC was examined. DC were derived from monocytes upon culture with interleukin (IL)-4 and granulocyte-macrophage colony-stimulating factor in the presence or absence of rapamycin (0.1-100 ng/mL). Surface phenotype and antigen uptake capacity of DC were assessed by flow cytometry. Immunostimulatory capacity was measured by mixed lymphocyte culture. RESULTS: Rapamycin reduced DC recovery and increased DC apoptosis. DC differentiated in the presence of rapamycin (rapa-DC) had increased expression of CD1a, CD1b, and CD1c and decreased expression of MHC I, MHC II, CD80, CD86, and CD40. Antigen uptake receptor expression (mannose receptor, CD32, CD91, CD46) was decreased, and receptor-mediated endocytosis of fluorescein isothiocyanate-dextran was markedly impaired in rapa-DC, as were fluid phase endocytosis of Lucipher Yellow and phagocytic activity of bacteria and dead or apoptotic cells. CD40 ligand-induced production of both IL-12 and IL-10 was reduced in rapa-DC, and allogeneic T lymphocyte responses were moderately impaired when rapa-DC were used as stimulator cells. Neither cyclosporine nor FK506 affected DC function. However, the effects of rapamycin on DC could be completely inhibited by a 10-fold excess of FK506 but not by up to 100-fold excess of cyclosporine. CONCLUSION: Rapamycin has a unique and profound inhibitory effect on DC function, which seems to be at least in part mediated by the FKBP immunophilins. PMID- 12544887 TI - Early differentiation between rejection and infection in liver transplant patients by serum and biliary cytokine patterns. AB - BACKGROUND: Differentiation between acute liver graft rejection and infection remains a clinical challenge during the early posttransplantation period. Although cytokines play a pivotal role in mediating allograft rejection, previous studies demonstrate that most cytokines are not specific for liver graft rejection or infections. However, other studies suggest that adhesion molecules and cytokines in bile reflect the immunologic activity within the liver more closely. Therefore, we postulated that by combining cytokine patterns in serum and bile, early recognition of acute liver graft rejection and differentiation from infectious complications can be improved. METHODS: We performed a prospective study in 45 patients who were monitored daily for clinical events and cytokine patterns in serum and bile during the first month after liver transplantation. RESULTS: Soluble intercellular adhesion molecule-1 (sICAM-1) in serum and interleukin-8 in bile were specifically increased at the onset of acute rejection (P<0.001), whereas serum soluble tumor necrosis factor-receptor II was also significantly increased in patients with infectious complications and serum interleukin-6 only in patients with rejection during infection. In 68% of patients with increased sICAM-1, acute rejection was diagnosed within 10 days, whereas rejection occurred in only 26% of patients with low serum levels of sICAM 1. In patients with increased sICAM-1, the relative risk for rejection was 4.8 (P=0.009). CONCLUSIONS: Cytokine patterns in bile do not provide rejection markers with higher specificity compared with serum cytokines. Daily monitoring of sICAM-1 in serum could identify patients at risk for rejection; therefore, acute liver graft rejection may be recognized earlier in those patients. PMID- 12544888 TI - Angioedema in pediatric liver transplant recipients under tacrolimus immunosuppression. AB - BACKGROUND: The authors report on their experience with food-induced angioedema in tacrolimus-immunosuppressed pediatric liver recipients. METHODS: Among 121 children treated with tacrolimus after liver transplantation, those who presented with angioedema are reported. RESULTS: Twelve children (10%) experienced angioedema related to food allergy while on tacrolimus. Mean ages at transplantation and angioedema were 1.3 years and 3.75 years, respectively. Angioedema occurred within a mean of 28 months from onset of tacrolimus. Eleven children experienced two or more angioedema attacks without consequences. One child presented with anaphylactic shock that caused postischemic cerebral damage. Besides eviction of food allergens, eight children were switched from tacrolimus to cyclosporine, whereas tacrolimus dosage was decreased in four. Reintroduction of food allergens was successfully performed only in those who were switched to cyclosporine. CONCLUSION: A causal relationship between tacrolimus and the occurrence of food-induced angioedema is suggested. The switch from tacrolimus to cyclosporine should be considered. PMID- 12544889 TI - Internet use by a multidisciplinary transplant clinic population. AB - BACKGROUND: This study was designed to determine patterns of patient Internet use and to define socioeconomic and other barriers that affect such use. METHODS: A 44-item questionnaire was distributed to patients seen in the transplant clinic during April 2001. RESULTS: A total of 170 patients completed the questionnaire. Fifty-six percent had access to the web. Seventy-two (42%) were African American, and 85 (50%) were Caucasian. Internet use was higher among Caucasian patients, but education level had the strongest correlation. Seventy-three percent of patients with a college education had Internet access compared with 38% of patients with a high school diploma. Twenty-two percent of patients used the Internet to find information about their doctor, and 35% searched for transplant survival results. CONCLUSIONS: Patients use the Internet to find information about their doctors and results of transplantation and other treatments. Physicians should consider establishing guidelines for use of Internet services by patients for education and communication. PMID- 12544890 TI - Severe acute renal failure after exposure to sirolimus-tacrolimus in two living donor kidney recipients. AB - BACKGROUND: We began a clinical trial in African Americans comparing sirolimus tacrolimus to standard immunosuppression. We report two African American male living donor kidney recipients who developed acute renal failure after exposure to sirolimus-tacrolimus. METHODS: Both patients received similar doses of sirolimus and tacrolimus to achieve target levels of 5 to 15 ng/mL and prednisone in tapering doses. Renal function and tacrolimus and sirolimus levels were systematically monitored. RESULTS: Although both kidneys functioned immediately, acute oliguric renal failure developed approximately 2 weeks after transplantation. Transplant kidney biopsy showed acute tubular necrosis in patient 2. Sirolimus-tacrolimus was then stopped in both patients. Both patients required temporary hemodialysis. Renal function returned 2 weeks later and was normal 2 months after transplantation on tacrolimus plus mycophenolate mofetil. CONCLUSION: Combination sirolimus-tacrolimus may cause nephrotoxicity in some patients by mechanisms that are presently unexplained. PMID- 12544891 TI - Association of glycoprotein B and immediate early-1 genotypes with human leukocyte antigen alleles in renal transplant recipients with cytomegalovirus infection. AB - BACKGROUND: Numerous risk factors for cytomegalovirus (CMV) infection or disease, or both, such as serostatus of donor and recipient, immunosuppressive regimen, or intensity of viral load, have been identified in renal transplant recipients. Additional parameters may be involved, notably, genetic variability of both host and virus, which could modulate the efficacy of the immune response. METHODS: Active CMV infection was analyzed retrospectively in 634 renal transplant recipients, according to human leukocyte antigen (HLA)-A, HLA-B, and HLA-DR alleles; CMV serostatus; presentation of the disease; and variations in the coding sequences of glycoprotein (g) B and IE1 proteins. RESULTS: Active infection occurred in 141 of 634 patients: seropositivity of the donor and the recipient were identified as risk factors. Patients carrying the HLA-A11, HLA A32, or HLA-DR11 allele developed active infection more frequently, whereas none of the patients with the HLA-B16 or HLA-B55 allele was actively infected. Significant independent associations between some genotypes and particular HLA alleles were observed: gB1 was more frequent in the HLA-A24 or HLA-B7 context and underrepresented in patients with HLA-DR11; gB2 was more frequent in HLA-A32 or HLA-DR11 carriers; and an increased frequency of gB3 was observed in the HLA-A29 context. Considering the IE1-2 genotype, increased frequency was noted for HLA-A3 carriers, whereas this type was underrepresented for patients with the HLA-DR11 allele. CONCLUSION: Data strongly suggest that differential presentation of polymorphic gB or IE peptides by HLA molecules or differential recognition by host CD8+ and CD4+ T lymphocytes, or both, should modulate immunologic response and then CMV pathogenesis in renal transplant patients. PMID- 12544892 TI - Posttransplant malaria: first case of transmission of Plasmodium falciparum from a white multiorgan donor to four recipients. PMID- 12544893 TI - The resident experience on trauma: declining surgical opportunities and career incentives? Analysis of data from a large multi-institutional study. AB - PURPOSE: The surgical resident experience with trauma has changed. Many residents are exposed to predominantly nonoperative patient care experiences while on trauma rotations. Data from a large multicenter study were analyzed to estimate surgical resident exposure to trauma laparotomy, diagnostic peritoneal lavage (DPL), and focused abdominal sonography for trauma (U/S). METHODS: Centers completed a self-report questionnaire on their institutional demographics, admissions, and procedure for a 2-year period (1998-1999). RESULTS: A total of 82 trauma centers that provide resident teaching were included. The included centers represent over 247,000 trauma admissions. The majority of trauma centers (65.9%) had > 80% blunt injury. Although all centers performed laparotomies, other results were more variable. For U/S, 24.2% performed none at all and 47.0% performed fewer than two U/S examinations per month. For DPLs, 3.8% performed none and 66.7% performed fewer than two per month. Assuming 1 night of 4 on call, the average surgical resident training at a trauma center performing > 80% blunt trauma has the potential to participate in only 15 trauma laparotomies, 6 diagnostic peritoneal lavages, and 45 ultrasound examinations per year. In addition, the resident will care for an average of 500 blunt trauma patients before performing a splenectomy or liver repair. CONCLUSION: Surgical resident experience on most trauma services is heavily weighted to nonoperative management, with a relatively low number of procedures, little experience with DPL, and highly variable experience with ultrasound. These data have serious implications for resident training and recruitment into the specialty. PMID- 12544894 TI - Charges and reimbursement at a rural level I trauma center: a disparity between effort and reward among professionals. AB - BACKGROUND: A Level I trauma center must provide immediate availability general (trauma) surgical expertise. In the current practice few patients require a general surgical procedure. The expertise of subspecialists may also be required and frequently these patients will require subspecialty operative care. We hypothesized that trauma surgeons would receive less reimbursement than their subspecialty colleagues despite a greater commitment of time and effort in taking care of the multiply-injured patient. METHODS: Three fellowship trained trauma surgeons were specifically hired to cover the trauma service for the year 2000. Professional billings, contribution to margin (reimbursement minus direct costs) of the trauma surgeons and subspecialists were obtained from the hospital financial information system. A surrogate for effort was assessed by the number of attending notes in the chart. A surrogate for complexity of care was assessed by the length of notes in the chart. Weekly time sheets assessed the percentage of time involved in the care of trauma patients. RESULTS: There were 344 patients cared for exclusively on the trauma service for the year 2000. The billing generated per patient was $1005 for the trauma surgeon, $5904 for the subspecialists, and $27,554 for the hospital. Orthopedics and radiology generated more professional billing on the trauma patients than the trauma surgeons. The trauma surgeons spent 52% of their weekly clinical activity in the care of trauma patients, yet this activity accounted for only 16% of their billings (the rest came from general surgery and ICU care). The effort and complexity of care provided by the trauma surgeons was significantly greater than the subspecialists. CONCLUSION: The Level I trauma service is a conduit for patients coming into the hospital that provides a significant remuneration to the subspecialty services. Trauma surgeons are able to bill much less than many of their subspecialty colleagues despite expending significantly greater amounts of time and effort in the care of these patients. Strategies for improved reimbursement for trauma surgeons must be devised or trauma surgery will suffer the same fate as other areas of surgery, losing our brightest and best to more financially sound subspecialty services such as radiology and orthopedics. PMID- 12544895 TI - Is there a relationship between trauma center volume and mortality? AB - BACKGROUND: The guidelines for Level I trauma center verification require 1,200 admissions per year. Several studies looking at the relationship between hospital volume and outcomes after injury have reached conflicting conclusions. The goal of our study was to examine the relationship between patient volume and outcomes (mortality and length of hospital stay) in California's trauma centers. METHODS: Data for patients >or= 18 years old admitted after injury (n = 98,245) to a Level I or II trauma center (n = 38) in 1998 and 1999 were obtained from the Patient Discharge Data of the State of California. Hospital volume was derived from the annual number of admissions per center, and covariates including age, sex, mechanism of injury, Injury Severity Score, and trauma center designation were analyzed. RESULTS: Hospital volume was not a significant predictor of death or length of hospital stay. More severely injured patients appeared to have worse outcomes at the highest volume centers. CONCLUSION: In our study, hospital volume was not a good proxy for outcome. Low-volume centers appeared to have outcomes that were comparable to centers with higher volumes. Perhaps institutional outcomes rather than volumes should be used as a criterion for trauma center verification. PMID- 12544896 TI - A twelve-year analysis of disease and provider complications on an organized level I trauma service: as good as it gets? AB - BACKGROUND: The development of trauma systems reduces preventable mortality and the measurement of standardized complications creates further opportunity for improvement in morbidity. The annual incidence of complications in a trauma population has been previously reported but the frequency change over time in a single institution is not well studied. METHODS: All patients who were hospitalized for more than 24 hours, who died, were admitted to the Intensive Care Unit (ICU) or Intermediate Care Unit (IMU), or were inter-facility transfers prospectively evaluated for 12 consecutive years. A total of 13,382 patients were studied (range, 862-1234 patients per year). Complication events were collected using 135 standardized definitions including disease and provider outcomes. RESULTS: The overall incidence of complications has remained stable over time. Provider events, disease events, and patients developing three or more complications have remained unchanged as well. Specific disease complications including pneumonia, deep vein thrombosis (DVT), and small bowel obstruction have fallen over time. Improvements in provider errors have also occurred. CONCLUSION: This data suggests that most complications have a finite threshold despite the use of a stable trauma staff, implementation of standardized protocols, and emphasis on consistency of practice. Further reductions will require new research for disease-related treatment and new strategies for consistency and error reduction rather than our current models of continuous quality improvement. PMID- 12544897 TI - Implementation of a tertiary trauma survey decreases missed injuries. AB - BACKGROUND: Missed injuries (MIs) adversely affect patient outcome and damage physician/institutional credibility. The primary and secondary surveys are designed to identify all of a patient's injuries and prioritize their management; however, MIs are prevalent in severely injured and multisystem trauma patients, especially when the patient's condition precludes completion of the secondary survey. We hypothesized that implementation of a routine tertiary trauma survey (TS) would reduce the incidence of MIs in a Level I trauma center. METHODS: In mid 1999, a TS form was created and TS documentation was mandated on all trauma intensive care unit (TICU) patients within 24 hours of admission. Patient data, including TS documentation and injury patterns, were concurrently recorded in an institutional trauma registry. Data were compared for patients admitted in 1997 to 1998 (PRE period) and 2000 to 2001 (POST period) using chi or Student's test. RESULTS: MIs decreased from 2.4% to 1.5% overall, and from 5.7% to 3.4% in TICU patients, after TS implementation. Patients with MIs were slightly older (49 vs. 45 years; > 0.05) and had higher Injury Severity Scores (21 vs. 10; < 0.05) than patients without MIs. Sixty percent of MI patients had brain injuries, 56% were admitted to the TICU, and 26% went directly from the emergency department to the operating room. The large majority of MIs in the POST period were detected in patients not undergoing timely TS. CONCLUSION: ICU patients-particularly brain injury victims and those undergoing emergent surgical procedures-appear to be at highest risk for MI. Implementation of a standardized TS decreased MIs by 36% in our Level I trauma center, and more timely TS would likely have further reduced MIs. A TS should be routine in trauma centers. PMID- 12544898 TI - Normal electrocardiography and serum troponin I levels preclude the presence of clinically significant blunt cardiac injury. AB - BACKGROUND: Uncertainty about the definition and diagnosis of blunt cardiac injury (BCI) leads to unnecessary hospitalization and cost while trying to rule it out. The purpose of this study was to examine whether the combination of two simple tests, electrocardiography (ECG) and serum troponin I (TnI) level, may serve as reliable predictors of BCI or the absence of it. METHODS: Over a period of 30 months (September 1999-February 2002), 333 consecutive patients with significant blunt thoracic trauma were followed prospectively. Serial ECG and TnI tests were performed routinely and echocardiography was performed selectively. Clinically significant BCI (SigBCI) was defined as the presence of cardiogenic shock, arrhythmias requiring treatment, or posttraumatic structural deficits. RESULTS: SigBCI was diagnosed in 44 patients (13%). Of 80 patients with abnormal ECG and TnI, 27 (34%) developed SigBCI. Of 131 with normal serial ECG and TnI, none developed SigBCI. Of patients with abnormal ECG only or TnI only, 22% and 7%, respectively, developed SigBCI. The positive and negative predictive values were 29% and 98% for ECG, 21% and 94% for TnI, and 34% and 100% for the combination of ECG and TnI. The admission ECG or TnI was abnormal in 43 of 44 patients with SigBCI. Only one patient had initially normal ECG and TnI and developed abnormalities 8 hours after admission. Forty-one patients without other significant injuries stayed 1 to 3 days in the hospital only to rule out SigBCI and could have been discharged earlier. Besides ECG and TnI, other independent risk factors of SigBCI were an Injury Severity Score > 15, the presence of significant skeletal trauma, and history of cardiac disease. CONCLUSION: The combination of normal ECG and TnI at admission and 8 hours later rules out the diagnosis of SigBCI. In the absence of other reasons for hospitalization, such patients can be safely discharged. PMID- 12544899 TI - Not so FAST. AB - BACKGROUND: Focused assessment with sonography for trauma (FAST) as a screening tool in the evaluation of blunt abdominal trauma will lead to underdiagnosis of abdominal injuries and may have an impact on treatment and outcome in trauma patients. METHODS: From October 2001 to June 2002, a protocol for evaluating hemodynamically stable trauma patients with suspected blunt abdominal injury (BAI) admitted to our institution was implemented using FAST examination as a screening tool for BAI and computed tomographic (CT) scanning of the abdomen and pelvis as a confirmatory test. At the completion of the secondary survey, patients underwent a four-view FAST examination (Sonosite, Bothell, WA) followed within 1 hour by an abdominal/pelvic CT scan. The FAST examination was considered positive if it demonstrated evidence of free intra-abdominal fluid. Clinical, laboratory, and imaging results were recorded at admission, and FAST examination results were compared with CT scan findings, noting the discordance. RESULTS: Patients with suspicion for BAI were evaluated according to protocol (n = 372). Thirteen cases were excluded for inadequate FAST examinations, leaving 359 patients for analysis. There were 313 true-negative FAST examinations, 16 true positives, 22 false-negatives, and 8 false-positives. Using CT scanning as the confirmatory test for hemoperitoneum, FAST examination had a sensitivity of 42%, a specificity of 98%, a positive predictive value of 67%, a negative predictive value of 93%, and an accuracy of 92%; chi analysis showed significant discordance between FAST examination and CT scan (5.85%, < 0.001). Six patients with false negative FAST examinations required laparotomy for intra-abdominal injuries; 16 patients required admission for nonoperative management of injury. Of the 313 true-negative FAST examinations, 19 patients were noted to have intra-abdominal injuries without hemoperitoneum and 11 patients were noted to have retroperitoneal injuries. CONCLUSION: Use of FAST examination as a screening tool for BAI in the hemodynamically stable trauma patient results in underdiagnosis of intra-abdominal injury. This may have an impact on treatment and outcome in trauma patients. Hemodynamically stable patients with suspected BAI should undergo routine CT scanning. PMID- 12544900 TI - Penetrating zone II neck injury: does dynamic computed tomographic scan contribute to the diagnostic sensitivity of physical examination for surgically significant injury? A prospective blinded study. AB - OBJECTIVE: The purpose of this study was to prospectively evaluate the utility of dynamic computed tomographic (CT) scanning as a diagnostic tool and adjunct to physical examination in the identification of surgically significant penetrating zone II neck injuries. METHODS: All patients older than 14 years of age who suffered penetrating zone II neck injuries were eligible for entry into the study protocol at an urban Level I trauma center. All patients that presented with signs of surgically significant injury on physical examination underwent immediate neck exploration. Patients that did not show signs of surgically significant injury were entered into the study protocol and underwent soft tissue dynamic CT scan (1/2-cm cuts, 250-mL oral contrast) of the neck after initial resuscitation. After CT scan, all patients entered into the study protocol underwent esophagography. After completion of radiologic assessment, all study protocol patients underwent surgical exploration of the neck. The patient's surgical team was blinded to results of the CT scan and esophagography before and during surgical exploration of the neck. RESULTS: During a 42-month period from May 1997 to March 2001, 42 patients were entered into the study protocol. Thirty six (86%) of the injuries were secondary to stab wounds and the rest were caused by gunshot wounds. Surgical exploration revealed four esophageal injuries, of which two (50%) were missed by CT scan. Esophagography missed the identical esophageal injuries, as did CT scan. Both of the missed esophageal injuries were secondary to stab wounds. Seven internal jugular vein injuries were diagnosed intraoperatively, of which four (57%) were diagnosed by CT scan. During the study period, all patients with carotid artery and tracheal injuries were diagnosed by physical examination and thus underwent immediate surgical exploration without study entry. CONCLUSION: Dynamic CT scan contributes minimally to the sensitivity of physical examination in the diagnosis of surgically significant penetrating zone II neck injury. Diagnosis of esophageal injuries with dynamic CT scan appears no better than esophagography. CT scan has greater sensitivity than physical examination for the diagnosis of jugular venous injuries; however, the majority of these injuries do not require identification or surgical intervention. PMID- 12544902 TI - Green fluorescent protein-adenoviral construct as a model for transient gene therapy for human cultured keratinocytes in an athymic mouse model. AB - BACKGROUND: The goal of gene therapy for cultured keratinocyte grafts is to accelerate growth and wound healing following engraftment without producing long term complications from the delivered gene. We studied a Green Fluorescent Protein-Adenoviral construct (GFP-ADV) to determine the characteristics of gene expression in human cultured keratinocyte grafts. METHODS: Twelve GFP-ADV grafts and twelve control grafts were transplanted to the flanks of 24 athymic mice. Mouse flanks were monitored with fluorescence-filtered microscopy and, on Day 21, were sectioned and stained with anti-human MHC Class I with H&E counterstaining. Real-time PCR was performed on graft biopsies for adenoviral DNA. RESULTS: Fluorescence decreased from Days 3 to 5 resulting in no difference between GFP ADV and control grafts from days 5 to 10. All grafts were positive for human MHC Class I with an epithelial architecture by H&E. Day 21 GFP-ADV grafts were negative for adenoviral DNA. CONCLUSION: The delivered gene was transiently expressed without the persistence of viral DNA, demonstrating the potential of adenoviral gene delivery for the improvement of wound healing without long-term adverse effects to the graft. PMID- 12544901 TI - Seven hundred fifty-three consecutive deaths in a level I trauma center: the argument for injury prevention. AB - BACKGROUND: The past century has seen improvement in trauma care, with a resulting decrease in therapeutically preventable deaths. We hypothesize that further major reduction in injury mortality will be obtained through injury prevention, rather than improvements in therapy. METHODS: Seven hundred fifty three deaths in an American College of Surgeons-verified, Level I trauma center were reviewed as they occurred. Deaths were classified as therapeutically not preventable, possibly preventable, or preventable. These charts were also reviewed for factors that might have prevented or lessened the severity of the injury. RESULTS: Mean age was 43, mean Glasgow Coma Scale score was 5, mean Revised Trauma Score was 4, mean Injury Severity Score was 41, and mean probability of survival was 0.25 (according to TRISS). Forty-six percent underwent cardiopulmonary resuscitation in the field, 52% died within 12 hours, 74% died within 48 hours, and 86% died within 7 days. Primary causes of death included central nervous system injury in 51%, irreversible shock in 21%, multiple injuries (shock plus central nervous system injury) in 9%, multiple organ failure/sepsis and other causes in 3%, and pulmonary embolus in 0.1%. Seven hundred one (93%) were classified as not preventable with a change in therapy, 32 (4.2%) were classified as potentially preventable with a change in therapy, and 20 were classified as preventable with a change in therapy (2.6%). Forty-six percent had cardiopulmonary resuscitation performed before or immediately on arrival to the hospital. Another 23% had vital signs present on arrival, but had a Glasgow Coma Scale score of 45% total body surface area thermal injury or healthy volunteers). Select cultures were pretreated with myosin light chain kinase inhibitors (ML-9). Permeability was assessed by migration of bovine serum albumin across cell monolayers. Cells were stained with rhodamine phalloidin and anti-ZO-1 antisera and examined by means of confocal microscopy. RESULTS: Burn serum significantly enhanced monolayer permeability to albumin, whereas pretreatment with ML-9 limited this effect. Control cells maintained cortical F-actin and peripheral ZO-1 distributions (1a, b), whereas burn serum induced transcellular F-actin stress fiber formation and a diffuse ZO-1 staining (2a, b). ML-9 prevented burn-induced actin rearrangements, but not the diffuse redistribution of ZO-1. CONCLUSION: These data demonstrate that endothelial F actin stress fiber formation and ZO-1 redistribution contribute to postburn loss of pulmonary endothelial monolayer integrity. Although myosin phosphorylation appears to be required for endothelial F-actin stress fiber formation, redistribution of actin-membrane anchoring proteins appears to be regulated independently after thermal injury. PMID- 12544904 TI - Role of downstream metabolic processing of proinflammatory fatty acids by 5 lipoxygenase in HL-60 cell apoptosis. AB - BACKGROUND: Proinflammatory eicosanoids formed from arachidonic acid (AA) by lipoxygenase (LO) and cyclooxygenase (COX) pathways have been shown to inhibit apoptosis in certain cell types. This study determined whether inhibition of LO and COX increased apoptosis in AA-treated HL-60 cells in vitro. METHODS: HL-60 cells were incubated with 50 micromol/L AA and an enzyme inhibitor (1-10 micromol/L) for COX, LO, 12-LO, and 5-LO for 12 hours. Flow cytometry was used to assess viability, apoptosis, and necrosis. Apoptosis was further assessed using terminal dUTP nick end-labeling and DNA fragmentation. RESULTS: The highest concentration of LO inhibitors, but not COX inhibitors, decreased viability and increased apoptosis and necrosis in the presence of exogenous AA. CONCLUSION: These results suggest that disruption of the metabolism of AA by LO, in particular 5-LO, decreases cell survival and increases apoptosis. Thus, downstream metabolic processing of AA by LO but not COX plays a critical role in the regulation of HL-60 cell apoptosis. PMID- 12544905 TI - Modulation of endotoxin-induced endothelial activity by microtubule depolymerization. AB - BACKGROUND: Endotoxin not only activates the Toll-mediated signaling pathway within endothelial cells that leads to neutrophil migration but also causes the polymerization of microtubules. The potential role of this polymerization event, however, is unknown. METHODS: Human umbilical vein endothelial cells stimulated with endotoxin were pretreated with or without the microtubule depolymerizing agent colchicine. Toll-mediated signaling events and protein production were in turn investigated by Western blot, gel shift, and enzyme-linked immunosorbent assay. Finally, neutrophil adhesion was assayed fluorometrically under the various conditions. RESULTS: Endotoxin led to activation of the various Toll mediated pathways, production of intercellular adhesion molecule-1 and interleukin-8, and subsequent neutrophil adhesion. Pretreatment with colchicine led to selective inhibition of anti-dual phosphorylated extracellular signal regulated kinase-1/2, anti-dual phosphorylated c-jun N-terminal kinase, and adaptor protein-1; selective enhancement of p38; and no effect on nuclear factor kappaB. This selective modulation of intracellular signaling resulted in attenuated intercellular adhesion molecule-1, interleukin-8 and prostaglandin E2 production, but enhanced cyclooxygenase-2 expression. As a result, microtubule disruption led to a significant reduction in neutrophil adhesion. CONCLUSION: Microtubule formation is essential to optimal endotoxin-induced intracellular signaling through anti-dual phosphorylated extracellular signal-regulated kinase 1/2, anti-dual phosphorylated c-jun N-terminal kinase, and adaptor protein-1. Failure of these signaling events is associated with a marked reduction in the formation of a proadhesive phenotype that may prove to be beneficial in modulating neutrophil recruitment during sepsis. PMID- 12544906 TI - Increased production of leukocyte microparticles with enhanced expression of adhesion molecules from activated polymorphonuclear leukocytes in severely injured patients. AB - BACKGROUND: Polymorphonuclear leukocyte (PMNL)-derived microparticles (MPs) have been recently reported as activators of vascular endothelium in vitro. The objectives of the present study were to evaluate the production of MPs in severely injured patients and to clarify the role of these MPs. METHODS: Thirty severely injured patients (mean Injury Severity Score of 27 +/- 11) and 21 healthy volunteers participated in the study. Blood samples were obtained serially at three time points: days 0 to 1, days 2 to 5, and days 6 to 12 after the trauma event. MP production, CD11b and CD62L expression on MPs, and oxidative activity in PMNLs were measured by flow cytometry in both the presence and absence of formylmethionyl-leucyl-phenylalanine. Expressions of CD11b and CD62L were differentially evaluated according to the size of the MPs (>or= or < 1.0 microm). Soluble E-selectin and thrombomodulin levels in blood, variables representative of systemic vascular endothelial damage, were also measured. RESULTS: Production of MPs with and without formylmethionyl-leucyl-phenylalanine and the oxidative activity in PMNLs (O ) were prominently increased on days 2 to 5 after trauma. CD62L expression was enhanced on MPs at all three time points, and CD11b expression was enhanced on MPs < 1.0 microm in diameter at all three time points. Soluble E2-selectin and thrombomodulin in blood did not change significantly between time points. CONCLUSION: Activated PMNLs enhance production of PMNL-derived MPs with increased adhesion molecule expression on days 2 to 5 after severe trauma. This response per se, however, may not progress to systemic vascular endothelial damage. PMID- 12544907 TI - Hypertonic saline resuscitation attenuates neutrophil lung sequestration and transmigration by diminishing leukocyte-endothelial interactions in a two-hit model of hemorrhagic shock and infection. AB - BACKGROUND: Hypertonic saline (HTS) attenuates polymorphonuclear neutrophil (PMN) mediated tissue injury after hemorrhagic shock. We hypothesized that HTS resuscitation reduces early in vivo endothelial cell (EC)-PMN interactions and late lung PMN sequestration in a two-hit model of hemorrhagic shock followed by mimicked infection. METHODS: Thirty-two mice were hemorrhaged (40 mm Hg) for 60 minutes and then given intratracheal lipopolysaccharide (10 microg) 1 hour after resuscitation with shed blood and either HTS (4 mL/kg 7.5% NaCl) or Ringer's lactate (RL) (twice shed blood volume). Eleven controls were not manipulated. Cremaster intravital microscopy quantified 5-hour EC-PMN adherence, myeloperoxidase assay assessed lung PMN content (2 1/2 and 24 hours), and lung histology determined 24-hour PMN transmigration. RESULTS: Compared with RL, HTS animals displayed 55% less 5-hour EC-PMN adherence (p = 0.01), 61% lower 24-hour lung myeloperoxidase ( p= 0.007), and 57% lower mean 24-hour lung histologic score ( p= 0.027). CONCLUSION: Compared with RL, HTS resuscitation attenuates early EC-PMN adhesion and late lung PMN accumulation in hemorrhagic shock followed by inflammation. HTS resuscitation may attenuate PMN-mediated organ damage. PMID- 12544908 TI - Alteration of the postinjury hyperinflammatory response by means of resuscitation with a red cell substitute. AB - BACKGROUND: Transfusion of stored packed red blood cells (PRBCs) has unintended effects beyond the desired results of increased oxygen delivery. A particular concern is the potential for lipid and cytokine mediators present in PRBCs to augment the postinjury inflammatory response that sometimes culminates in multiple organ failure. Through the use of a polymerized human hemoglobin (PolyHeme), we have been able to measure the inflammatory response in patients resuscitated with minimal exposure to banked components in the early postinjury period. METHODS: Critically injured patients requiring urgent transfusion were resuscitated with either PRBCs or PolyHeme in the early postinjury period. Proinflammatory cytokines (interleukin [IL]-8 and IL-6), counterregulatory cytokines (IL-10 and IL-11), and markers of endothelial injury (soluble intercellular adhesion molecule and soluble E-selectin) were serially measured. RESULTS: Increases in IL-8, IL-6, and IL-10 were greater in patients resuscitated with PRBCs. IL-11 plasma levels were largely below the level of detection of the assay. There was no difference in markers of endothelial injury. CONCLUSION: Consistent with concerns about the immunoinflammatory response to transfusion of PRBCs, we observed exaggerated levels of three markers associated with adverse outcome. The clinical significance of these findings with respect to the development of multiple organ failure awaits further study. PMID- 12544909 TI - Correlation of intracellular organisms with quantitative endotracheal aspirate. AB - BACKGROUND: The presence of intracellular organisms (ICOs) in polymorphonuclear cells obtained from respiratory secretions is a possible method for rapid diagnosis of ventilator-associated pneumonia. We correlated ICOs with quantitative endotracheal aspirate (QA) in intubated patients. METHODS: Consecutive intubated patients in the surgical intensive care unit had respiratory samples obtained every 2 days until extubation. Two thresholds for ICOs and quantitative culture were examined. Sensitivity, specificity, and positive and negative predictive values were calculated using QA as reference. RESULTS: One hundred one samples were obtained from 35 patients. Colony counts >or= 100,000 were found in 34 samples; 60 samples had colony counts >or= 10,000. Antibiotic use did not affect the sensitivity or specificity of ICOs. Sensitivity of ICOs was 39% to 85%, and specificity was 82% to 97%. Positive predictive value was 70% to 96%, and negative predictive value was 50% to 91%. CONCLUSION: ICOs provide a quick method for establishing the presence of a significant bacterial load in the respiratory tract. Accuracy of ICOs in predicting a positive QA is not affected by concurrent antibiotics. PMID- 12544910 TI - Two New York City hospitals' surgical response to the September 11, 2001, terrorist attack in New York City. AB - BACKGROUND: We describe the surgical response of two affiliated hospitals during the day of, and week following, the September 11th, 2001 terrorist attack at the World Trade Center in New York City. The city of New York has 18 state designated regional trauma centers that receive major trauma victims. The southern half of Manhattan is served by a burn center, two regional trauma centers, and a community hospital that is an affiliate of one of the regional trauma centers. This report accounts for the surgical response by a regional trauma center (Hospital A, located 2.5 miles from the World Trade Center) and its affiliate hospital (Hospital B, located 5 city blocks from the World Trade Center) on September 11th when two commercial jets crashed into the Twin Towers at the World Trade Center mall. METHODS: Hospital A maintained a concurrent log of patients received during the first 5 hours, the first day, and the first week after the disaster which was kept by the Surgical Triage Officer. The trauma registry completed and verified this data by September 18th. Hospital B collected its data by hand counting and verification by chart review. Both hospitals, A and B, had established disaster plans that were implemented. RESULTS: Nine hundred eleven patients were received by two affiliated hospitals from the World Trade Center attack. Seven hundred seventy six patients (85%) were walking wounded, sustaining mild inhalation and eye irritant injuries. One hundred thirty five (15%) were admitted with 18 (13%) of these undergoing surgery. Twenty two of the 23 transfers were from the community hospital to specialized orthopedic or burn centers. Of the 109 patients admitted to Hospital A, 30 were to the surgical service. The mean ISS score of these patients was 12. There were 4 deaths (within minutes of arrival at the hospital) and 6 delayed deaths (day 1-14). Excluding walking wounded and DOAs, the critical mortality rate was 37.5% overall. CONCLUSION: The September 11th, 2001, terrorist attack in New York City, involving two commercial airliners crashing into the World Trade Center, led to 911 patients received at two affiliated hospitals in lower Manhattan. One hospital is a regional trauma center and one was an affiliate community hospital. Eighty five percent of the patients received were walking wounded. Of the rest, 13% underwent surgical procedures with an overall critical mortality rate of 37.5%. PMID- 12544911 TI - Sheriff's surgeon's alert: a trauma surgeon's responsibility. PMID- 12544912 TI - The changing face of trauma management and its impact on surgical resident training. AB - BACKGROUND: The management of trauma patients has become increasingly nonoperative, especially for solid abdominal organ injuries. However, the Residency Review Committee (RRC) still requires an operative trauma experience deemed essential for graduating general surgical residents. The purpose of this study was to review the trauma volume and mix of patients at two trauma centers and determine the major operative trauma cases available to residents involved in the care of these patients. METHODS: A retrospective chart review was conducted at the two trauma centers used by the Michigan State University surgery residency. Both of the trauma centers are American College of Surgeons verified. Surgical residents are involved with the care of every trauma patient at each of the hospitals. Cumulative data were collected and analyzed from January 1, 1997, through December 31, 1999. Age, gender, mechanism of injury (blunt vs. penetrating), Injury Severity Score, length of stay, operative interventions, and patients managed nonoperatively were reviewed. RESULTS: There were 434 patients selected for this study from 2,340 patients admitted to the trauma services. Male patients accounted for 66% of patients and female patients accounted for 34% of patients. Blunt trauma was the mechanism in 89% of patients, with penetrating trauma accounting for the other 11% of patients. Of the total number of patients, motor vehicle crashes accounted for the majority of cases, 325 of 434 (75%). Overall, 85% (370 of 434) of patients were managed without an index trauma surgical procedure according to RRC guidelines. Only 14.7% (64 of 434) of patients underwent operative intervention that qualified as index trauma surgical cases identified by the RRC. The spleen and small bowel were the two most commonly injured organs found at laparotomy. Nonoperative intervention of many patients with solid abdominal organ injuries did not meet the operation requirements expected by the RRC. CONCLUSION: Our residency program had 10 graduating chief residents over the 3-year time period. With only 64 operative trauma cases, this yields an average of 6.4 trauma cases per resident. This falls significantly short of the 16-case minimum requirement in trauma surgery established by the RRC. The operative trauma requirements established by the RRC for graduating residents may be unattainable in many residency programs because of the high incidence of blunt trauma and the changing patterns of trauma management. PMID- 12544913 TI - Contrast material extravasation on contrast-enhanced helical computed tomographic scan of blunt abdominal trauma: its significance on the choice, time, and outcome of treatment. AB - BACKGROUND: Contrast-enhanced helical computed tomographic (CT) scan of blunt abdominal trauma is valuable for detecting contrast material extravasation (CME). The aims of this study were to determine its significance and investigate factors associated with the choice, time, and outcome of management. METHODS: CT scans of 32 consecutive trauma patients who had CME were reviewed for the sources of CME, types of CME, flat inferior vena cava, and multiple abdominal injuries. The medical records were reviewed for demographics, systolic blood pressure, Injury Severity Score (ISS), choice of management, time interval between CT scan and intervention, and outcome of intervention. RESULTS: Systolic blood pressure < 100 mm Hg was the most important factor (p = 0.0064) that failed observational therapy. When proceeding to intervention treatment, patients with a flat inferior vena cava (1.6 +/- 1.1 hours) had a significantly shorter time interval between CT scan examination and intervention when compared with those with a normal cava (10.9 +/- 16.0 hours) ( p= 0.0124). The mortality rate after intervention treatment was 18.8%. High ISS, uncontained CME in the extraperitoneum, and multiple abdominal injuries were important risk factors. After adjusted for ISS and multiple abdominal injuries, the risk of dying from extraperitoneal CME remained significant when compared with intraperitoneal CME (adjusted odds ratio, 82.26; 95% confidence interval, 1.06-6,363.17). CONCLUSION: Termination of observational therapy was appropriate for trauma patients who had CME and systolic blood pressure < 100 mm Hg. The coexistence of a flat inferior vena cava and CME was associated with early intervention treatment. Despite early intervention, the mortality rate was 18.8%. High ISS and multiple abdominal injuries were important factors, but the risk of dying from uncontained extraperitoneal CME was 82 times the risk of dying from intraperitoneal CME. PMID- 12544914 TI - The role of interventional radiology in patients requiring damage control laparotomy. AB - BACKGROUND: Interventional angiography has been used as a less invasive alternative to surgery to control hemorrhage resulting from trauma. This retrospective study analyzed the role of interventional radiology in patients requiring damage control laparotomy. METHODS: Twenty patients underwent damage control laparotomy between January 1994 and May 2001. Eight of the 20 patients also underwent angiographic evaluation and treatment before or after the damage control laparotomy. RESULTS: Three patients underwent angiography before damage control laparotomy, because a large, pelvic retroperitoneal hematoma was seen on computed tomographic scan, and the amount of intraperitoneal blood seemed insufficient to account for the magnitude of the patient's hemodynamic instability. Five patients underwent angiography after damage control laparotomy. The indication was a nonexpanding retroperitoneal hematoma in three patients, a nonexpanding hepatic hilar hematoma in one patient, and a hepatic injury associated with cirrhosis in one patient. Lumbar artery injuries were identified and treated by embolization in three patients. Four of the eight patients who underwent both damage control laparotomy and angiography survived. CONCLUSION: Angiography before damage control laparotomy may be indicated to control retroperitoneal pelvic hemorrhage in hemodynamically unstable patients who have insufficient intraperitoneal blood loss to account for their hemodynamic instability. Angiography after damage control laparotomy should be considered when a nonexpanding, inaccessible hematoma is found at operation in a patient with a coagulopathy. PMID- 12544915 TI - Effect of a chitosan-based hemostatic dressing on blood loss and survival in a model of severe venous hemorrhage and hepatic injury in swine. AB - BACKGROUND: Hemorrhage is a leading cause of death from trauma. An advanced hemostatic dressing could augment available hemostatic methods. We studied the effects of a new chitosan dressing on blood loss, survival, and fluid use after severe hepatic injury in swine. METHODS: Swine received chitosan dressings or gauze sponges. Standardized, severe liver injuries were induced. After 30 seconds, dressings were applied and resuscitation initiated. Blood loss, hemostasis, resuscitation volume, and 60-minute survival were quantified. RESULTS: Posttreatment blood loss was reduced ( p< 0.01) in the chitosan group (264 mL; 95% confidence interval [CI], 82-852 mL) compared with the gauze group (2,879 mL; 95% CI, 788-10,513 mL). Fluid use was reduced ( p= 0.03) in the chitosan group (1,793 mL; 95% CI, 749-4,291) compared with the gauze group (6,614 mL; 95% CI, 2,519-17,363 mL). Survival was seven of eight and two of even in the chitosan and gauze groups ( p= 0.04), respectively. Hemostasis was improved in the chitosan group ( p= 0.03). CONCLUSION: A chitosan dressing reduced hemorrhage and improved survival after severe liver injury in swine. Further studies are warranted. PMID- 12544916 TI - Man-in-the-barrel syndrome after hemorrhagic shock. PMID- 12544917 TI - Blunt abdominal trauma resulting in urinothorax from a missed uretero-pelvic junction avulsion: case report. PMID- 12544918 TI - Traumatic injury of the pelvis causing compression and acute critical stenosis of a common iliac artery stent with resultant critical leg ischemia: successful treatment with transbrachial balloon angioplasty: case report. PMID- 12544919 TI - Thrombosis of the portal venous system after splenectomy for trauma. PMID- 12544920 TI - Hypokalemia in acutely injured children: a benign laboratory abnormality. PMID- 12544921 TI - Politically correct discussions at surgical meetings. PMID- 12544922 TI - Use of diagnostic peritoneal lavage (DPL) as a tool for minimizing nontherapeutic laparotomy rate. PMID- 12544923 TI - Functional outcome in trauma patients with spinal injury. PMID- 12544924 TI - Ultrasound imaging of the intervertebral disc. AB - STUDY DESIGN: In vitro ultrasound imaging of dog intervertebral discs was performed. OBJECTIVE: To determine the reliability of ultrasound imaging in the detection of structural changes associated with disc pathology. SUMMARY OF BACKGROUND DATA: Little work has been done to evaluate the potential applications of ultrasound in the imaging of intervertebral discs. Initial in vitro studies, however, have indicated that ultrasound imaging is capable of producing images of the disc that contain a high degree of structural information. METHODS: Explanted lumbosacral discs from 13 nonchondrodystrophic dogs, mean age 5 years and 8 months, were stripped of all surrounding tissues and scanned using ultrasound before being sectioned and photographed. The ultrasound images were graded according to criteria chosen to reflect progressive stages of disc degeneration, allowing correlation with the grading scale used to assess the photographic images of the discs. Grades assigned to each disc were compared using Cronbach's alpha to determine the reliability of the images obtained using ultrasound. RESULTS: The results for the anterior anulus fibrosus produced an alpha value of 0.924; those for the nucleus pulposus produced a value of 0.821; whereas those for the posterior anulus fibrosus produced a value of 0.882. Where the grade given to the ultrasound image did not match those given in visual assessment, the disparity was never greater than one grade. Ultrasound images of several discs demonstrated echo patterns that matched, in both location and appearance, real structural defects identifiable on the sectioned discs. CONCLUSIONS: Ultrasound images of intervertebral discs relate well to their pathologic condition. In addition, ultrasound is able to locate specific pathologic defects. PMID- 12544925 TI - Complement membrane attack complexes in pathologic disc tissues. AB - STUDY DESIGN: Complement membrane attack complexes were located in lumbar spine disc tissues by immunohistochemistry. Their occurrence was compared in control discs obtained from organ donors (CD), discs showing a normal macroscopic anatomy, samples of intervertebral disc herniations (DH), and intervertebral discs found to be degenerated by discography, but not herniated (DD). OBJECTIVE: To look for a possible role of complement activation, specifically complement membrane attack complexes, an end product of the classic immune complex-mediated complement activation pathway, in disc pathophysiology. SUMMARY OF BACKGROUND DATA: Recent immunohistochemical and biochemical studies suggest a possible role for immune complexes, as observed by immunohistochemical location and biochemical assay of immunoglobulins M and G in intervertebral disc pathophysiology. Immune complexes may trigger complement activation and ultimately cell lysis. There are, however, currently no reports on complement activation in disc tissues, although immune (antigen-antibody) complexes have been demonstrated. Such immune complexes have been reported to occur on or near to disc cells in DH tissues. METHODS: Thin frozen sections of disc tissue from CD (n = 9 discs), DH (n = 58 discs), and DD (n = 11 discs) were cut and then immunostained with a monoclonal antibody to the complement membrane attack complex (C5b-9) using avidin-biotin complex (ABC) immunostaining. The presence or absence of complement membrane attack complex immunoreactivity was compared in the various subtypes of DH and also with preoperative duration of radicular pain. RESULTS: Complement membrane attack complexes could be observed in none of the CDs studied. In contrast, in more than one third of both the DH (21/58, 36.2%) and the DD (4/11, 36.4%), immunoreactivity to complement membrane attack complexes could be observed in disc cells. In DD discs, immunoreactivity to complement membrane attack complexes was most often present in anulus fibrosus samples (5/13, 38.5%). With respect to subtype of DH, complement membrane attack complexes were observed in 19 of 36 sequestrated discs (52.8%), 1 of 16 extrusions (6.3%), and 1 of 6 protrusions (16.7%). Complement membrane attack complexes were more often present with shorter pain duration (P= 0.03), but showed no relation to age. Disc cells often showed a heavy staining pattern for complement membrane attack complexes, suggesting an abundance of these complexes lodged in the membrane of the cells. CONCLUSIONS: The predominant presence of complement membrane attack complexes in sequestrated disc tissue could suggest a role in DH tissue-induced sciatica. Possibly immune (antigen-antibody) complexes, reported in previous studies, trigger the classic pathway of complement activation, with complement membrane attack complexes as the final product. Complement membrane attack complexes also appear to have some as yet undefined role in degenerated nonherniated disc tissue, with a predominant presence in the anulus fibrosus cells of such discs. PMID- 12544926 TI - Cerebral spinal fluid pressure: effects of body position and lumbar subarachnoid drainage in a canine model. AB - STUDY DESIGN: This study used in vivo an model of subarachnoid cerebrospinal fluid pressure measurement. OBJECTIVES: To examine the relation between subarachnoid cerebrospinal fluid pressure in the cervical and lumbar spine and varying body positions, and to test the hypothesis that increasing body inclination and lumbar subarachnoid drainage decreases cervical cerebrospinal fluid pressures. SUMMARY OF BACKGROUND DATA: Cerebrospinal fluid leaks are a recognized complication of anterior or posterior cervical surgery. Conflicting opinion exists regarding the benefits of postoperative patient positioning and lumbar subarachnoid drainage. METHODS: Subarachnoid cerebrospinal fluid pressure of 7 beagles was monitored via two angiocatheters attached to pressure transducers inserted into the subarachnoid space through laminectomies at C3 and L4. Pressure measurements were taken when body position was inclined to 30 degrees, 60 degrees, and 90 degrees. A lumbar durotomy was performed to simulate the effects of lumbar subarachnoid drainage. The body was repositioned to 90 degrees, and pressure was measured. RESULTS: As inclination increased from 0 degrees to 90 degrees, the mean cervical cerebrospinal fluid pressure significantly decreased. The mean lumbar subarachnoid pressure significantly increased as inclination increased from 0 degrees to 90 degrees. Lumbar durotomy plus repositioning to 90 degrees resulted in a significant reduction in cervical cerebrospinal fluid subarachnoid pressure, with pressures dropping by 46%. Lumbar cerebrospinal fluid subarachnoid pressure dropped to zero after lumbar durotomy plus repositioning to 90 degrees. CONCLUSIONS: Cerebrospinal fluid pressures in the subarachnoid space of both the cervical and lumbar spines are affected by changes in body position. Both patient positioning and lumbar drainage may be important in reducing cervical cerebrospinal fluid pressure, and may reduce the occurrence of cerebrospinal fluid leaks after primary dural repair in the neck. PMID- 12544927 TI - Micro-computed tomography evaluation of trabecular bone structure on loaded mice tail vertebrae. AB - STUDY DESIGN: A micro-computed tomography (CT) study of the trabecular bone structure on loaded mice tail vertebral bodies was conducted. OBJECTIVE: To depict and characterize changes in the trabecular bone structure of mice tail vertebral bodies after in vivo application of static compressive load. SUMMARY OF BACKGROUND DATA: Static compressive loading leads to significant structural changes in murine tail intervertebral discs, such as disorganization of the anulus fibrosus, increase in apoptosis, and associated loss of cellularity. Wolff's Law suggests that alterations in spinal loading will also influence the architecture of the adjacent vertebral bodies. Because of biomechanical and biologic interdependencies between the disc and vertebra, these tissues should be considered simultaneously when investigating the etiology of degenerative spinal conditions. METHODS: Mice tail discs between the ninth and 10th caudal vertebrae were compressed in vivo for 7 days with static axial loads using external fixators. Micro-CT scans of the vertebral bodies were performed at an isotropic resolution of 18 microm, to obtain trabecular bone structural parameters. Random effects models were used to evaluate statistical significance of these parameters in different compressed conditions. RESULTS: With loading, the connectivity density of the trabecular network increases significantly. After a period of in vivo recovery on load removal, the trabeculae become more rod-like; corresponding changes such as disorganization of the anulus fibrosus and loss of nuclear and inner-anular cellularity are also seen. CONCLUSIONS: In vivo compressive loading leads to significant architectural changes within vertebral bodies. These observations may be helpful in understanding the pathologic processes and the chronology of degenerative spinal conditions. PMID- 12544928 TI - A cross-sectional study correlating degeneration of the cervical spine with disability and pain in United kingdom patients. AB - STUDY DESIGN: A prospective cross-sectional study was conducted. OBJECTIVE: To determine the relation between degeneration of all the joints in the cervical spine, and to identify the effects of litigation on pain and disability levels in trauma and nontrauma patients. SUMMARY OF BACKGROUND DATA: The link between spinal degeneration and patient symptoms remains controversial. Two recent similar studies present conflicting results concerning the association between spinal degeneration and symptomatology. These two studies, performed in different countries, did not consider the impact of impending litigation on self-reported pain and disability levels. The role of the uncovertebral and facet articulations has hitherto not been investigated. METHODS: Radiographic and questionnaire data from 180 consecutive patients with neck pain were collected. Neck pain severity was measured using two time-dependent scales. All patients completed the Neck Disability Index (NDI) and questions about chronicity, etiology, and associated litigation. The radiographs were evaluated for the number of levels of degeneration and the severity of degeneration in the discs as well as the uncovertebral and facet articulations. RESULTS: In this study, 71 patients (40.57%) reported neck pain as a result of injury. However, only 5.1% had associated litigation. There was no statistically significant difference in pain severity or disability levels between the patients with and those without cervical degeneration. According to the findings, the number of levels of cervical degeneration and the severity of degeneration in the discs, facets, and uncovertebral joints are not related to the levels of pain and disability. Patients reporting neck pain caused by injury had a tendency (P = 0.055) to more pain during the preceding week and significantly more disability (P < 0.001). Significant differences included pain intensity (P < 0.025), reading (P < 0.001), headaches (P < 0.025), ability to drive (P < 0.01), and concentration (P < 0.01). Women reported significantly more pain (P < 0.01) and disability (P < 0.001) than men, but did not have more degeneration in any of the joints. CONCLUSIONS: In patients with neck pain, there is no difference in reported pain and disability levels between those with and those without evidence of cervical spine degeneration. Patients whose neck pain is the result of trauma report significantly more pain and disability than nontrauma patients. This is not because of more spinal degeneration or overriding litigation issues. PMID- 12544929 TI - Donor site morbidity after anterior iliac crest bone harvest for single-level anterior cervical discectomy and fusion. AB - STUDY DESIGN: This retrospective, questionnaire-based investigation evaluated iliac crest bone graft (ICBG) site morbidity in patients having undergone a single-level anterior cervical discectomy and fusion (ACDF) procedure performed by a single surgeon (T.J.A.). OBJECTIVE: To evaluate acute and chronic problems associated with anterior ICBG donation, particularly long-term functional outcomes and impairments caused by graft donation. SUMMARY OF BACKGROUND DATA: Anterior cervical discectomy and fusion procedures frequently use autologous anterior ICBG to facilitate osseous union. Although autologous ICBG offers several advantages over alternative grafting materials, donor site morbidity can be significant. Acute and chronic complications of donor sites have been reported, yet there are currently no reports of long-term functional outcomes after autologous anterior ICBG donation after single-level ACDF. METHODS: A questionnaire was mailed to 187 consecutive patients who were retrospectively identified to have undergone autologous anterior ICBG harvest for single-level ACDF between 1994 and 1998. The questionnaire divided items into symptomatic (acute and chronic) and functional assessments. Patients answered yes, no, or not applicable; pain was assessed with a Visual Analogue Scale (VAS). RESULTS: Surveys were completed either by mail or follow-up telephone interview by 134 patients (71.6%). Average follow-up was 48 months (range, 24-72 months). Acute symptoms were reported at the following rates: ambulation difficulty, 50.7%; extended antibiotic usage, 7.5%; persistent drainage, 3.7%; wound dehiscence, 2.2%; and incision and drainage, 1.5%. The chronic symptom questionnaire demonstrated a high degree of satisfaction with the cosmetic result (92.5%). Pain at the donor site was reported by 26.1% of patients with a mean VAS score of 3.8 in 10, and 11.2% chronically use pain medication. Twenty-one patients (15.7%) reported abnormal sensations at the donor site, but only 5.2% reported discomfort with clothing. A unique functional assessment revealed current impairments at the following rates: ambulation, 12.7%; recreational activities, 11.9%; work activities, 9.7%; activities of daily living, 8.2%; sexual activity, 7.5%; and household chores, 6.7%. CONCLUSIONS: A large percentage of patients report chronic donor site pain after anterior ICBG donation, even when only a single level ACDF procedure is performed. Moreover, long-term functional impairment can also be significant. Patients should be counseled regarding these potential problems, and alternative sources of graft material should be considered. PMID- 12544930 TI - Early reconstruction failures after multilevel cervical corpectomy. AB - STUDY DESIGN: A retrospective analysis of graft and plate complications after multilevel anterior cervical corpectomy and fusion (ACF) attributed to spondylosis, stenosis, and ossification of posterior longitudinal ligament was conducted. OBJECTIVE: To identify factors contributing to graft and plate complications in this population. SUMMARY OF BACKGROUND DATA: Biomechanical factors contributing to the increased morbidity associated with plated multilevel ACF were evaluated. METHODS: Graft- and/or plate-related complications were retrospectively reviewed in 33 patients undergoing two-level ACF reconstructions and in seven patients having three-level ACF reconstructions performed with iliac crest grafting and instrumentation with a fixed-plated design (cervical spine locking plate). Neurologic status was assessed before surgery and after surgery using both the Nurick Grading Scale and modified JOA (Japanese Orthopaedic Association) Score. The patients were observed an average of 31.4 months after surgery. The follow-up included lateral flexion and extension radiographs and a neurologic examination. RESULTS: Two of the 33 patients undergoing two-level fusions available for long-term follow-up after surgery developed reconstruction failures. All of the remaining fusions were successful, demonstrated by lateral flexion and extension radiographs. Seven patients had plated three-level corpectomy reconstructions. Five of the seven who had anterior-only reconstruction failed. DISCUSSION: A two-level ACF reconstruction is reliable with an anterior strut graft and fixed screw plate construct. A three-level ACF reconstruction is not reliably achieved with an anterior-only construct. The construct failures may be attributed in part to the fixed-plated design being used, as well as the long lever arm of the construct. CONCLUSION: There is a 6% failure rate after fixed-plated (cervical spine locking plate) two-level ACF reconstruction but a 71% failure rate after three-level fixed-plated ACF reconstruction. Future consideration should be given to simultaneous posterior fusion. PMID- 12544931 TI - Clinical characteristics of chronic back pain as a function of gender and oral opioid use. AB - STUDY DESIGN: A cross-sectional analysis of data derived from patients with chronic spinal pain undergoing evaluation at a multidisciplinary pain treatment center was conducted. OBJECTIVE: To determine whether pain severity, psychological status, and physical disability differed as a function of gender and opioid use, and whether the clinical correlates of opioid use differed in women and men with chronic back pain. SUMMARY OF BACKGROUND DATA: Gender differences in the experience of pain have been widely reported. For example, in the general population, several chronic pain conditions are more prevalent among women than among men, and many experimental studies demonstrate lower pain thresholds and tolerances among women. In addition, recent evidence from studies of experimental and acute clinical pain suggests that responses to analgesic medications may differ in women and men. METHODS: The sample consisted of 240 patients (35% women) with low back, upper back, or neck pain undergoing evaluation for treatment at a multidisciplinary pain center. The patients were classified as opioid or nonopioid users on the basis of self-report and medical record review. All the patients completed a battery of clinical assessments, including measures of pain severity, psychological adjustment, self-reported disability, functional tasks, and pain tolerance. Analyses were conducted to examine clinical variables as a function of gender and opioid use. RESULTS: The results indicated that opioid use was associated with greater self-reported disability and poorer function in both women and men. However, the association of opioid use with affective distress differed between women and men. The women using opioids showed lower affective distress, whereas the opioid-using men reported greater affective distress. Opioid use was not associated with pain severity, although the women reported greater pain than men. CONCLUSIONS: These findings indicate that both opioid use and gender are significant predictors of clinical status of patients with chronic spinal pain. More interesting, these two variables interact because opioid use was associated with increased affective distress among the men, but the reverse was true for the women. In addition, the women reported greater pain severity, which is consistent with some previous findings. Potential explanations for these findings are presented, and the practical implications are discussed. PMID- 12544932 TI - Pelvic lordosis and alignment in spondylolisthesis. AB - STUDY DESIGN AND OBJECTIVES: Pelvic morphology and lumbopelvic lordosis were measured on standing radiographs of 75 patients with greater than 10% L5-S1 spondylolytic spondylolisthesis. The findings were compared with those of 75 volunteers to determine significant differences between the two groups. SUMMARY OF BACKGROUND DATA: Etiology of isthmic (lytic) spondylolisthesis remains uncertain. Causation appears to be multifactorial. The relationship between pelvic morphology and spondylolisthesis deserves additional study. METHODS: Both groups had a standing lateral radiograph of the thoracolumbar spine and pelvis taken that included both hips. Three radiographic angles for pelvic morphology (pelvisacral, pelvic incidence, and pelvic lordosis) were measured by two observers. Each offered similar reliability. Measurement of the pelvic lordosis angle by the pelvic radius technique required fewer steps. It also allowed calculation of the combined angles comprising both the pelvic morphology component for lordosis (the constant pelvic lordosis angle) and the lordosis in the lumbar spine (the variable lumbar lordosis from T12-S1) that should complement the fixed pelvic lordosis (the complementary lumbopelvic lordosis). Mean values and statistical correlations were then computed for each group and compared. RESULTS: The mean slippage for patients was 30% (range, 11-85%), with 34 patients (45%) having Grade I slips, 32 (43%) having Grade II slips, and nine (12%) having Grade III and IV slips. The mean measurements between patients and volunteers were significantly different (P < 0.01) for lumbar lordosis, pelvic lordosis, and lumbopelvic lordosis. Subgroups of patients with increasingly larger slips (Grade I-III) had significantly smaller mean angles for pelvic lordosis. CONCLUSIONS: The pelvic and lumbopelvic parameters studied were different in patients compared with controls. The contribution of the pelvis to lordosis was significantly smaller in the subgroups of patients with increasingly larger grades of spondylolisthesis. Pelvic morphology may play a role in the development of spondylolisthesis. Measurement of the combined lumbar and pelvic (lumbopelvic) lordosis on standing radiographs is important. PMID- 12544933 TI - Discriminant validity and relative precision for classifying patients with nonspecific neck and back pain by anatomic pain patterns. AB - STUDY DESIGN: Secondary analysis of a previously described cohort of prospective, consecutive patients with acute neck or low back pain referred to outpatient rehabilitation was performed. OBJECTIVE: To estimate discriminant validity and relative precision of two classification procedures (first visit vs multiple visit) in discriminating short-term pain intensity and perceived disability outcomes. SUMMARY OF BACKGROUND DATA: Centralization and noncentralization are pain responses used to classify patients and predict outcomes. Different time frames have been proposed for operationally defining these responses, which are problematic for comparing outcomes across clinical trials. Classifying patients according to pain response observed from initial examination (first visit) and over time (multiple visits) influences prevalence within categories and interpretation of classification usefulness, which merits further investigation. METHODS: Patients with acute onset of nonspecific neck or low back pain referred to two outpatient physical therapy clinics completed body pain diagrams, pain intensity ratings, and disability questionnaires at initial evaluation, during each visit, and at discharge. Therapists collected data enabling patient classification on initial examination and throughout treatment. Differences in pain and disability from intake to discharge from rehabilitation across classification categories were used to assess discriminant validity. Relative precision was estimated by determining ratios of analysis of covariance F values between classification procedures for pain and disability. RESULTS: Both classification procedures discriminated categories for change in pain and disability. The multiple-visit classification procedure was more precise for discriminating outcomes than the first-visit classification procedure. CONCLUSION: Multiple-visit classification of patients into specific pain pattern subgroups is recommended when pain intensity and disability outcomes are of interest. PMID- 12544934 TI - Oblique reformation in cervical spine computed tomography: a new look at an old friend. AB - STUDY DESIGN: Cervical spine computed tomograms were evaluated for neural foraminal stenosis in both the standard axial plane and the oblique reformatted plane. OBJECTIVE: To assess whether oblique reformation of cervical spine computed tomograms reduces interobserver variability in the evaluation of neural foraminal stenosis. SUMMARY OF BACKGROUND DATA: Radiographic assessment of neural foraminal stenosis is subjective, may vary among observers, and can affect surgical planning. METHODS: The cervical spine images from 19 patients with various degrees of neural foraminal stenosis were reformatted in an oblique plane perpendicular to the long axis of the right and left neural foramens. Seven independent observers graded the degree of foraminal stenosis (none, mild [1 25%], moderate [26-75%], or severe [>75%]) and their confidence level (definite, probable, possible) on both the axial images and the oblique reformations. RESULTS: The ages of the 12 male (mean, 67.5 +/- 13.24 years) and 7 female (mean, 62.7 +/- 14.79 years) patients ranged from 39 to 83 years. Interobserver variability was assessed with chi2 analysis. Rates of agreement on degree of stenosis (chi2 = 19.94; df= 9; P< 0.02) were significantly higher for oblique reformations. Confidence ratings also were significantly higher for oblique reformations (chi2 = 18.19; df= 7; P< 0.02). CONCLUSIONS: Oblique reformation of cervical spine images significantly reduces the degree of interobserver variability and increases observer confidence in the assessment of neural foraminal stenosis. Oblique reformations should be considered in the routine evaluation of neural foraminal stenosis. PMID- 12544935 TI - The relation of thoracic and lumbar fracture configuration to the development of late deformity in childhood spinal cord injury. AB - STUDY DESIGN: A retrospective clinical observational study was conducted. OBJECTIVE: To assess the relation of spinal fracture type and its magnitude of distortion to subsequent long-term development of late spinal deformity in childhood onset spinal cord injury. SUMMARY OF BACKGROUND DATA: In this study, 76 adults who sustained spinal cord injury during childhood were examined clinically and radiographically alongside a retrospective review of case notes and radiographs. METHODS: The nature of the spinal injury and the progression of its displacement were defined from radiographs taken immediately after injury, then at 4 months and at 1 year. Eventual adult spinal deformity was defined from standardized erect long-plate radiographs. Scoliosis, kyphosis, and lordosis were measured using Cobb's method. RESULTS: There was no statistically significant difference in the severity of scoliosis, kyphosis, or lordosis between traumatic and nontraumatic injuries, nor between patients with and those without radiologically visible bony injury. Of the 14 patients with traumatic thoracic and lumbar injuries who had undergone no surgical intervention, 10 (71%) showed development of major scoliotic curves that did not include the fracture site. The patients with no angular displacement at the fracture site after 1 year went on to experience the development of more severe scoliosis (mean, 66 degrees) than those who had displaced fractures (mean, 38 degrees). In five, a low kyphotic curve and a compensatory lordosis above it developed. CONCLUSIONS: There is no evidence that the bony injury to the vertebral column itself in the child with spinal cord injury influences the development of late scoliosis or lordosis, but it may influence any eventual kyphosis. PMID- 12544936 TI - Vitamin D deficiency and chronic low back pain in Saudi Arabia. AB - STUDY DESIGN: Initial assessment involved 360 patients (90% women and 10% men) attending spinal and internal medicine clinics over a 6-year period who had experienced low back pain that had no obvious cause for more than 6 months. The patients ranged in age from 15 to 52 years. OBJECTIVES: To investigate the contribution of vitamin D deficiency as a cause for idiopathic chronic low back pain, to find a simple and sensitive test for screening patients with low back pain for vitamin D deficiency, and to determine the correlation between the vitamin deficiency and pain. METHODS: A biochemical assay of serum calcium, phosphate, alkaline phosphatase, and 25-hydroxy vitamin D level was performed before and after treatment with vitamin D supplements. RESULTS: Findings showed that 83% of the study patients (n = 299) had an abnormally low level of vitamin D before treatment with vitamin D supplements. After treatment, clinical improvement in symptoms was seen in all the groups that had a low level of vitamin D, and in 95% of all the patients (n = 341). CONCLUSIONS: Vitamin D deficiency is a major contributor to chronic low back pain in areas where vitamin D deficiency is endemic. Screening for vitamin D deficiency and treatment with supplements should be mandatory in this setting. Measurement of serum 25-OH cholecalciferol is sensitive and specific for detection of vitamin D deficiency, and hence for presumed osteomalacia in patients with chronic low back pain. PMID- 12544937 TI - Functional outcome in trauma patients with spinal injury. AB - STUDY DESIGN: A retrospective data analysis of all trauma patients admitted the Helicopter Emergency Medical Service was performed. OBJECTIVE: To assess the long term outcome of trauma patients with spinal injuries using Functional Independence Measure scores. SUMMARY OF BACKGROUND DATA: Mortality after severe multiple trauma is well documented. However, evaluating morbidity in survivors of multiple trauma is complex, and less information is available regarding functional outcome. There are very few systems that can effectively predict the outcome for patients sustaining multiple trauma with spinal injuries. The Functional Independence Measure scoring system, which is easy to use, can be used to assess disability after hospital discharge, and may also be used to predict the long-term outcome for patients after spinal injuries. METHODS: The records of 1500 trauma patients admitted over a 6-year period by the Helicopter Emergency Medical Service were examined. All patients with documented injuries to the spinal column were selected for study. The distribution and pattern of spinal injury, the injury severity score, and the radiologic findings were determined for each patient, along with clinical outcome measures at 1 year using Functional Independence Measure scores. RESULTS: Among the 1500 trauma patients, 263 patients (17.5%) (195 men and 68 women; mean age, 37 years; range, 3-92 years) had sustained an injury to the spinal column. Mortality (70/263; 27%) was significantly higher (P < 0.02) in these patients than in those without spinal injury (247/1237; 20%). Injury severity scores higher than 16 were found in 96 patients (55%). The median Functional Independence Measure score was 40 on admission, 86 at discharge from the hospital, 113 at 3 months, 119 at 6 months, and 124 at 12 months. There was significant correlation between discharge Functional Independence Measure (FIM) scores (FIM = 86) and 12-month FIM scores (FIM = 124) (P < 0.01). CONCLUSIONS: Most of the patients had poor initial Functional Independence Measure scores, but there was significant improvement by 12 months. Discharge FIM scores were a good indicator for functional outcome at one year. PMID- 12544938 TI - Recent advances in disc cell biology. AB - STUDY DESIGN: There have been many advances over the past decade in understanding and experimentally modulating biologic aspects of intervertebral disc cell function. An overview of the current state of this biologic research is presented. OBJECTIVES: To provide clinicians with a review of important recent advances in biologic studies of the disc and their implications for potential disc therapies. SUMMARY OF BACKGROUND DATA: Historically, anatomic, biochemical, radiologic, and biomechanical studies of the intervertebral disc formed the foundation on which our understanding of disc function was built. Magnetic resonance imaging techniques that allowed viewing of soft tissue components of the disc further advanced imaging capabilities. METHODS: Recent publications are reviewed. RESULTS: Experimental approaches over the past decade have enabled researchers to look more critically at disc cell function. This is important because disc cell function produces the extracellular matrix components of the disc, which, in turn, shape the disc's subsequent physiologic and biomechanical functions. New approaches to the study of disc cell function, methods to manipulate disc cells, studies of intact discs and disc nutrition, vertebral endplate structure and function, tissue engineering, gene therapy, and the potential of stem cells in disc therapy are reviewed and discussed. CONCLUSIONS: Many believe that disc degeneration has a cellular basis. New research is helping us better understand healthy, aging, and degenerating discs. Modern methods to manipulate and modulate disc cell function open exciting and challenging new therapeutic possibilities for future biologic treatments of disc degeneration. PMID- 12544939 TI - Monitoring scoliosis surgery with combined multiple pulse transcranial electric motor and cortical somatosensory-evoked potentials from the lower and upper extremities. AB - STUDY DESIGN: A retrospective case review was performed. OBJECTIVE: To assess the value, rapidity, and safety of combined multiple-pulse transcranial electric stimulation motor-evoked potential and somatosensory-evoked potential monitoring during scoliosis surgery. SUMMARY OF BACKGROUND DATA: Leg somatosensory-evoked potentials can miss motor deficits, and a 50% amplitude warning criterion can produce false alarms. METHODS: For this study, 33 scoliosis surgeries in neurologically normal patients under propofol/fentanyl anesthesia omitting neuromuscular blockade were monitored with four-extremity multiple-pulse transcranial electric stimulation muscle motor-evoked potentials and cortical somatosensory-evoked potentials. Instead of amplitude criteria, parallel (same direction) change was used to identify systemic alteration and nonparallel (one- or two-limb) deterioration to identify focal neurologic compromise. Clinical observation and intraoperative electroencephalography were used to assess adverse effects. RESULTS: Instantaneous motor-evoked potentials and rapidly reproducible cortical somatosensory-evoked potentials provided comprehensive feedback every 0.8 to 6.7 minutes (median, 2.4 minutes) without adverse effects. Parallel (systemic) changes without alarm or deficit included motor-evoked potential fading or temporary loss and leg somatosensory-evoked potential amplitudes below 50% of initial, maximum, or median intraoperative values in 10% to 37% of the cases. Three nonparallel changes occurred: 1) abrupt bilateral leg somatosensory evoked potential 20% to 30% reduction without motor-evoked potential change during instrumentation resolving spontaneously over 30 minutes, with transient postoperative sensory symptoms; 2) right-arm somatosensory-evoked potential and motor-evoked potential reduction during hyperabduction restored after repositioning, without deficit; 3) abrupt bilateral leg motor-evoked potential loss preceding 30% to 60% somatosensory-evoked potential reduction during derotation rapidly restored after instrumentation release, without deficit. CONCLUSIONS: In neurologically normal patients, the combined methods are safe and rapid, and could improve the sensitivity and specificity of scoliosis monitoring. Arm controls facilitate differentiation between systemic alterations and focal neurologic compromise. PMID- 12544940 TI - Symptomatic spinal arachnoid cysts: report of two cases with review of the literature. AB - STUDY DESIGN: Two unusual cases of intradural extramedullary noncommunicating arachnoid cysts are presented, with a review of literature. OBJECTIVE: To assess surgical excision results in the reversal of symptoms and neurologic deficits. SUMMARY OF BACKGROUND DATA: Most patients harboring arachnoid cysts are asymptomatic. Noncommunicating intradural extramedullary arachnoid cysts are more rare than communicating intradural extramedullary cysts. Noncommunicating intradural extramedullary arachnoid cysts are a very rare cause of spinal cord compression. Rarely, these cysts can present with bizarre symptoms, such as angina. MATERIALS AND METHODS: The first case, a 75-year-old woman, presented with progressive paraparesis. The second case, a 40-year-old man, presented with signs and symptoms of angina, with mild spasticity in the lower limbs. In both cases, magnetic resonance imaging revealed compression of the spinal cord as the etiology of presentation. The first patient is the second oldest patient in the literature with an idiopathic spinal arachnoid cyst. The second case has an unusual presentation that has not previously been reported in the literature. RESULTS: Both cases underwent surgical excision of the arachnoid cyst, with resolution of symptoms and reversal of neurologic deficit. CONCLUSIONS: Our experience indicates that complete recovery from an arachnoid cyst is possible after surgical excision, even when symptoms are present for a long duration, even in the elderly. PMID- 12544941 TI - Myopericytoma of the thoracic spine: a case report. AB - STUDY DESIGN: A case of myopericytoma of the thoracic spine is reported. OBJECTIVE: To report a recently described and extremely rare soft tissue neoplasm in the previously unreported location of the thoracic spine. SUMMARY OF BACKGROUND DATA: Myopericytoma is a recently described soft tissue neoplasm with perivascular myoid differentiation. All cases have been reported in the subcutaneous and superficial soft tissues of the extremities. This represents the first reported case of this unusual lesion in the spine. METHODS: A patient with a 3-month history of progressive weakness of the arms and legs was found to have a lytic lesion of T3. The lesion was surgically treated by curettage followed by a course of radiation. Presenting symptoms, imaging studies, histologic findings, and related literature are reviewed in this study. RESULTS: Histologic evaluation of sections of the surgically removed lesion revealed a concentric, periluminal proliferation of cells with prominent positive smooth muscle actin immunoreactivity. These findings are consistent with the diagnosis of a myopericytoma, a lesion not previously reported in the spine. CONCLUSIONS: This study shows the presence of a lesion in the thoracic spine only previously described in the soft tissue of the extremities. The diagnosis of myopericytoma should be included, along with hemangiopericytoma, in the differential diagnosis of lytic lesions of the spine. PMID- 12544942 TI - Horner's syndrome caused by a thoracic dumbbell-shaped schwannoma: sympathetic chain reconstruction after a one-stage removal of the tumor. AB - STUDY DESIGN: A case of Horner's syndrome caused by a thoracic dumbbell-shaped schwannoma is reported. OBJECTIVES: To report a rare case of a mediastinal dumbbell-shaped schwannoma as a cause of Horner's syndrome and to show the result of intercostal nerve grafting for sympathetic chain reconstruction after resection of the sympathetic nerve. SUMMARY OF BACKGROUND DATA: It has been reported that approximately 10% of neurogenic mediastinal tumors extend through the neural foramen into the spinal canal, creating a dumbbell shape. Although the most frequent causes of Horner's syndrome are tumors, a dumbbell-shaped schwannoma has rarely been described as a cause of the syndrome. Moreover, there have been no previous reports that primary sympathetic chain reconstruction has been performed with an intercostal nerve graft after resection of the sympathetic nerve with the tumor. METHODS: A 48-year-old woman was diagnosed with a mediastinal tumor by routine chest radiography. The patient had right-sided Horner's syndrome, the signs of which she had not noticed. Surgical resection of the dumbbell-shaped tumor was performed in a one-stage combined resection of both the intraspinal and the mediastinal component of the tumor. Primary sympathetic chain reconstruction was also performed with an intercostal nerve graft. RESULTS: The tumor was resected completely, and no recurrence of the tumor was observed 1 year after the operation. Blepharoptosis and anhidrosis on the right side of her face and upper limb gradually improved after surgery, and compensatory oversweating on the left side eventually improved. In bright illumination, the right pupil diameter was 3.5 mm and the left was 5 mm after surgery; the right pupil measured 4 mm and the left measured 5 mm 1 year after the operation. CONCLUSIONS: A mediastinal dumbbell-shaped schwannoma has rarely been described as a cause of Horner's syndrome. Primary sympathetic nerve reconstruction with an intercostal nerve was shown to be useful after resection of the sympathetic nerve involved in the tumor. PMID- 12544943 TI - Piriformis syndrome diagnosed by cauda equina action potentials: report of two cases. AB - STUDY DESIGN: Evaluation of recorded potentials from the lumbar spine after stimulation of the peroneal nerve in patients with piriformis syndrome is reported. OBJECTIVES: To report a new method of diagnosis of piriformis syndrome. SUMMARY OF BACKGROUND DATA: Clinically, the Freiberg and Pace signs were used to diagnose this entity. Among imaging examinations, computed tomography and magnetic resonance imaging have been used. An electrophysiological method using H reflex has also been reported. However, the best method for diagnosis of piriformis syndrome has yet to be clearly established. METHODS: After having an epidural electrode inserted into the lumbar spine, patients lay on their back. Cauda equina action potentials were recorded with the epidural electrode at L3/4 after stimulation of the peroneal nerve at the fibular head. As a control, the potentials were recorded with the hip and knee extended. Potentials were then also recorded with the hip flexed, and then with the hip flexed and internally rotated to stress the sciatic nerve by the piriformis muscle. RESULTS: We observed changes in amplitudes and latencies of recorded potentials. Amplitude decreased to about 70% of the control value in one case and to 67% in the other. On the nonaffected side, however, this amplitude did not decrease below 90%. The latency was also delayed on the affected side, but not on the nonaffected side. CONCLUSIONS: Recording of evoked potentials from the lumbar spine with stress to the affected leg was useful for diagnosing piriformis syndrome. PMID- 12544944 TI - Prenatal sonographic diagnosis of the congenital dislocated spine: a case report. AB - STUDY DESIGN: A case of congenital dislocated spine diagnosed by prenatal sonography is reported. OBJECTIVES: To report the first documented case of prenatal diagnosis, to point out the characteristic features of prenatal sonography of congenital dislocation of the spine, and to show the results of a careful management of birth in cases of congenital dislocated spine. SUMMARY OF BACKGROUND DATA: The congenital dislocated spine consists of an anterior defect in formation of the vertebrae with a displaced vertebra and angulation of the neural canal. Instability and the consequent possibility of neurologic damage are the major features of this rare entity. Early intervention is essential to prevent an irreparable neurologic deficit. METHODS: Pre- and postnatal imaging was reviewed. Early postnatal treatment is described. RESULTS: Prenatal diagnosis of the congenital dislocated spine can be detected with sonography and confirmed by radiography and magnetic resonance imaging. CONCLUSIONS: Prenatal diagnosis of the congenital dislocated spine is of the utmost importance for parental counseling and obstetrical management. PMID- 12544946 TI - A histologic and functional study on cauda equina adhesion induced by multiple level laminectomy. AB - STUDY DESIGN: A histologic and functional study on cauda equina adhesion induced by multiple-level laminectomy was performed using rats. OBJECTIVE: To assess the relation between the extent of laminectomy and histologic and functional changes of the cauda equina induced by laminectomy. SUMMARY OF BACKGROUND DATA: It has been demonstrated that laminectomies may induce cauda equina adhesion, transiently or permanently. However, the relation between the extent of laminectomy and histologic and functional changes of the cauda equina induced by laminectomy is still unclear. METHODS: Sprague-Dawley rats (250-300 g) were used. They were divided into four groups: no-laminectomy group (n = 5) and single, double, and triple level laminectomy groups (n = 20, respectively). At 24 hours and 1, 3, and 6 weeks after laminectomy, rats in each group were killed, and specimens were obtained from L4 to L6. The histologic changes were assessed using the classification of Yamagami. Walking durations on treadmill test and ascending spinal nerve-evoked potential were assessed in laminectomy groups before laminectomy, then weekly for 1 to 6 weeks after laminectomy. RESULTS: With the greater extent of laminectomy, the decrease in cauda equina adhesions was more delayed. There was no difference in walking durations and ascending spinal nerve evoked potential between pre- and postlaminectomy. CONCLUSIONS: The extent of laminectomy was related to recovery time of induced cauda equina adhesion. However, a delay in recovery from cauda equina adhesion may not produce a recognizable delay in recovery from clinical symptoms. PMID- 12544947 TI - Factors affecting the pullout strength of self-drilling and self-tapping anterior cervical screws. AB - STUDY DESIGN: A biomechanical comparison of 12-, 14-, and 16-mm self-tapping and self-drilling screws in cadaveric cervical spines was performed. OBJECTIVE: This study compared self-drilling and self-tapping screw designs used in anterior cervical locking plates in terms of insertion torque and pullout strength. In addition to screw design, the effects of screw length (12, 14, and 16 mm) and bone mineral density were examined. METHODS: A total of 201 screws were implanted and tested. Nineteen cadaver spine specimens (C2 through C7) were radiographed and their bone mineral density measured. In each vertebral level, two screws of the same length were implanted and their insertion torque measured. One of each screw design (self-drilling or self-tapping) was randomly placed in each vertebra for side-by-side comparison. Vertebrae then were individually potted and mounted on an MTS machine for accurate measurement of maximum pullout strength. RESULTS: Pullout strength was strongly related to screw length ( = 0.0002). There was no significant difference in pullout strength between the self-drilling and self tapping screw designs at any length ( = 0.9352). A significant correlation ( 0.10). The area under the receiver operating characteristic curve for the spinous process method was found to be 0.980, as compared with 0.662 for the Cobb angle method, for the measurement of pseudarthrosis. CONCLUSIONS: Measurement of the change in distance between spinous processes is more reproducible and accurate than the Cobb method for making the diagnosis of pseudarthrosis. The authors believe that the measurement of distances between spinous processes on lateral flexion-extension radiographs should be used as a method for evaluating radiographic fusion in patients with pseudarthrosis. PMID- 12544956 TI - Point of view. PMID- 12544957 TI - Reliability and diagnostic accuracy of the clinical examination and patient self report measures for cervical radiculopathy. AB - STUDY DESIGN: A blinded, prospective diagnostic test study was conducted. OBJECTIVES: To assess the reliability and accuracy of individual clinical examination items and self-report instruments for the diagnosis of cervical radiculopathy, and to identify and assess the accuracy of an optimum test-item cluster for the diagnosis of cervical radiculopathy. SUMMARY OF BACKGROUND DATA: Although cervical radiculopathy remains largely a clinical diagnosis, the reliability and diagnostic accuracy of clinical examination items, individually or in combination, for cervical radiculopathy is largely unknown. METHODS: Patients with suspected cervical radiculopathy or carpal tunnel syndrome received standardized electrophysiologic examination of the symptomatic upper quarter followed by a standardized clinical examination by physical therapist examiners blinded to diagnosis. Diagnostic properties were assessed using a neural impairment reference criterion standard. RESULTS: The study involved 82 patients. More than two thirds of 34 clinical examination items had reliability coefficients rated at least fair or better, and 13 items had likelihood ratio point estimates above 2 or below 0.50. A single diagnostic test item cluster of four variables was identified and produced a positive likelihood ratio point estimate of 30.3. The 95% confidence intervals for all likelihood ratio point estimates in this study were wide. CONCLUSIONS: Many items of the clinical examination were found to be reliable and to have acceptable diagnostic properties, but the test item cluster identified was more useful for indicating cervical radiculopathy than any single test item. Upper limb tension Test A was the most useful test for ruling out cervical radiculopathy. Further investigation is required both to validate the test item cluster and to improve point estimate precision. PMID- 12544958 TI - The reliability and concurrent validity of the scoliosis research society-22 patient questionnaire for idiopathic scoliosis. AB - STUDY DESIGN: Outcome study to determine response distribution, internal consistency, reproducibility, and concurrent validity of the Scoliosis Research Society-22 (SRS-22) health-related quality-of-life (HRQL) questionnaire. OBJECTIVES: Further refinement of an HRQL questionnaire specific for idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: Previous experience with the original and modified SRS HRQL questionnaires suggested a need for further refinement and more complete validation. METHODS: The SRS-22 and Short Form 36 (SF-36) HRQL questionnaires were mailed to 83 previously surveyed postoperative idiopathic scoliosis patients. RESULTS: Fifty-eight (70%) patients returned the first set of questionnaires. Their average age at surgery was 14.6 years, and their average follow-up interval since surgery was 10.8 years. Fifty-one (88%) of the 58 returned the second set of questionnaires an average of 28 days later. The psychometric attributes of the instruments were comparable: score distribution, SRS-22 56.9% ceiling and 1.7% floor, SF-36 79.3% ceiling and 1.7% floor; internal consistency (Cronbach alpha), SRS-22 0.92 to 0.75, SF-36 0.91 to 0.36; and reproducibility (intraclass correlation coefficient), SRS-22 0.96 to 0.85, SF-36 0.92 to 0.61. Concurrent validity, determined by Pearson Correlation Coefficients between SRS-22 and SF-36 domains, was 0.70 or greater ( < 0.0001) for 17 relevant comparisons. CONCLUSION: The SRS-22 HRQL questionnaire is reliable with internal consistency and reproducibility comparable to SF-36. In addition, it demonstrated concurrent validity when compared to SF-36. It is shorter and more focused on the health issues related to idiopathic scoliosis than SF-36. PMID- 12544959 TI - Scoliosis research society-22 patient questionnaire: responsiveness to change associated with surgical treatment. AB - STUDY DESIGN: Prospective observational consecutive case series. OBJECTIVES: To determine the responsiveness to change of the Scoliosis Research Society-22 (SRS 22) patient questionnaire after surgical treatment. SUMMARY OF BACKGROUND DATA: The SRS-22 outcomes questionnaire is the outgrowth of prior research that led to modifications and improvements of the original SRS questionnaire. METHODS: Fifty eight patients with an average age of 16 years and an average Cobb size of 63 degrees enrolled. They were tested preoperatively and at 3- (within 4 months), 6- (5-8 months), 12- (9-16 months), and 24-month (22-36 months) intervals postoperatively using the SRS-22 outcomes questionnaire. The paired Student test with multiple comparison adjustment was used to test significance of change score over time. Only patients with data on both time points of interest were included in the analysis. RESULTS: Self-image was significantly improved at 3 months ( < 0.0001) and maintained improvement through 24 months. Function was significantly decreased at 3 months ( < 0.0001), but returned to baseline by 6 months. Pain was significantly worse at 3 months ( = 0.0099), but was significantly less at 6 ( = 0.0011), 12 ( < 0.0001), and 24 ( = 0.0037) months when compared to 3 months. CONCLUSION: The SRS-22 questionnaire is responsive to changes in the postsurgical period. PMID- 12544960 TI - Discrimination validity of the scoliosis research society-22 patient questionnaire: relationship to idiopathic scoliosis curve pattern and curve size. AB - STUDY DESIGN: Prospective observational study. OBJECTIVE: To determine the discriminant validity of the Scoliosis Research Society-22 (SRS-22) health related quality-of-life (HRQL) questionnaire for idiopathic scoliosis patients based on curve pattern and curve size. SUMMARY OF BACKGROUND DATA: An important psychometric attribute of the HRQL questionnaire is the capacity to discriminate between and among patients with differing condition severity. METHOD: Three study groups, with subjects ranging in age from 10 to 16 years, were set up: a control group (C) of 19 persons with scoliosis suspected but unproven with an average age of 13 years; a nonsurgical group (NS) of 68 patients with an average age of 14 years and an average scoliosis (Cobb) of 30 degrees; and a presurgical group (PS) of 32 patients with an average age of 14 years and an average scoliosis of 61 degrees. The NS group was subdivided into untreated (NSU) (n = 54) and braced (NSB) patients (n = 14). Also created were subgroups of the NS and PS groups having similar curve sizes: nonsurgical similar (NSS) and presurgery similar (PSS). Statistical significance of the SRS-22 domain scores by comparison groups was tested by ANOVA. Relationships between the SRS-22 questionnaire and scoliosis deformity measurements were calculated using the Pearson Correlation Coefficients. RESULTS: There were no significant differences between the control (C) and nonsurgical (NS) groups. The presurgical group (PS) scored significantly lower in the pain and self-image domains than the C or NS groups and lower than group C in the mental health domain. A subgroup of the nonsurgical group (NSS) with curves comparable to the presurgical subgroup (PSS) had significantly better self-image scores. There were no significant correlations between SRS-22 scores and curve pattern. For persons with scoliosis, curve size (Cobb) correlated with pain, self-image, and function; trunk asymmetry correlated with self-image. CONCLUSION: The SRS-22 HRQL questionnaire successfully discriminated among persons with no scoliosis or moderate scoliosis and large scoliosis. It did not discriminate among patients with single, double, or triple curves. Self-image and, to a lesser extent, pain and function domain scores correlated with radiographic and trunk asymmetry severity. The SRS-22 HRQL questionnaire may be useful in choosing nonsurgical versus surgical treatment in borderline cases. PMID- 12544961 TI - Point of view. PMID- 12544962 TI - Rasch analysis of the Roland disability questionnaire. AB - STUDY DESIGN: A cross-sectional survey using patient questionnaires was conducted. OBJECTIVE To assess whether the Roland Disability Questionnaire satisfies the Rasch model including unidimensionality and item separation. SUMMARY OF BACKGROUND DATA: The Roland Disability Questionnaire, the most widely used patient-assessed measure of health outcome for back pain, has undergone several evaluations for its measurement properties including reliability, validity, and responsiveness. However, there is no published work relating to the underlying dimensionality of the instrument and the extent to which individual items contribute to the construct of physical disability resulting from low back pain. METHODS: Patients entering a randomized controlled trial of exercise, manipulation, and usual management for back pain completed a questionnaire that included the Roland Disability Questionnaire. The Winsteps program was used to assess whether the Roland Disability Questionnaire fits the Rasch model. Item fit was assessed using the Infit and Outfit statistics. RESULTS: The Roland Disability Questionnaire was completed by 1008 (90%) of the patients taking part in the trial. Most of the items in the Roland Disability Questionnaire contribute to a single underlying construct. However, four items had poor Outfit statistics, suggesting that they do not contribute sufficiently to the scale hierarchy. Several items positioned around the middle of the hierarchy are not sufficiently distinct in terms of difficulty. There were very few items positioned at the extremes of the hierarchy. CONCLUSIONS: The Roland Disability Questionnaire largely satisfies the Rasch model for unidimensionality. However, the instrument could be improved through the removal of poorly fitting items and the addition of items at the upper and lower points of the scale hierarchy. The distribution of Roland Disability Questionnaire scores should be carefully considered before statistical testing is undertaken. Rasch transformed scores can be used to deal with deficiencies in the scale hierarchy. PMID- 12544963 TI - Intraoperative measurements of nerve root blood flow during discectomy for lumbar disc herniation. AB - STUDY DESIGN: Nerve root blood flow was intraoperatively measured before and after discectomy for lumbar disc herniation and compared with clinical features. OBJECTIVE: To investigate the relation between nerve root blood flow changes and symptoms associated with lumbar disc herniation. SUMMARY OF BACKGROUND DATA: Several authors have reported that recovery of intraneural blood flow, which restores the supply of oxygen and other nutrients to the nerve tissue, is strongly related to the rapid improvement of nerve function after discectomy for lumbar disc herniation. However, no previous study has quantitatively assessed blood flow in the human nerve root. METHODS: Nerve root blood flow was monitored in 21 patients with lumbar disc herniation using laser Doppler flowmetry (ALF 21 N; ADVANCE, Tokyo, Japan) during discectomy. Possible correlations were investigated between the blood flow rates and the following clinical features: age, duration of sciatica, presence or absence of neurologic deficits, latency to pain relief, and morphology of herniated discs. RESULTS: The blood flow rate in 16 patients who reported immediate relief after discectomy was much greater than in 5 patients whose pain was not relieved immediately after surgery (141% 8%, = 0.0364). The increase in the blood flow rate after discectomy was five times greater in patients with neurologic deficits than in patients without neurologic deficits (158% 36%, = 0.0638). CONCLUSIONS: The results of the current study suggest that immediate relief from pain and resolution of neurologic deficits soon after surgery are the result of early recovery from nerve root ischemia after discectomy, and that ischemia caused by mechanical nerve root compression is mainly related to the mechanisms underlying sciatic pain production and neurologic deficits. PMID- 12544964 TI - Fluoroscopically assisted pedicle screw fixation for thoracic and thoracolumbar injuries: technique and short-term complications. AB - STUDY DESIGN: Retrospective review of charts and fluoroscopic, radiographic, and computed tomography images for 252 screws (41 patients). OBJECTIVES: To describe a fluoroscopically assisted technique of thoracic pedicle screw insertion and to determine the technique's safety, short-term complications, and accuracy. SUMMARY OF BACKGROUND DATA: Pedicle screw instrumentation is safe and effective for lumbar spine stabilization, but evidence of its accuracy and complications in the thoracic spine is lacking. METHODS: Between 1997 and 2000, 41 consecutive patients with unstable cervicothoracic, thoracic, and thoracolumbar spine injuries were admitted to a regional, level 1 trauma center. These patients underwent posterior spine arthrodesis and pedicle screw instrumentation placed intraoperative multiplanar fluoroscopic imaging. Pedicle screw placement accuracy was determined by review of postoperative computed tomography scans. Instrumentation failure prevalence was determined by review of fluoroscopic images, postoperative radiographs, and computed tomography scans. Neurologic, infectious, vascular, and gastrointestinal complications were determined by review of operative reports, hospital records, and clinic notes. RESULTS: Of the 126 (50%) thoracic screws (22 patients) with postoperative computed tomography scans, 16 (12.7%) penetrated the pedicle cortex (4 [2.4%] medially and 13 [10.3%] laterally) and 7 (5.6%) penetrated the vertebral body. There were four surgical complications: one instrumentation failure (broken rod, no loss of correction), one deep infection, and two superficial infections. The infections resolved with irrigation, debridement, and intravenous antibiotics. During the 6- to 24-month follow-up period, there were no neurologic, vascular, or gastrointestinal complications. CONCLUSIONS: Cervicothoracic, thoracic, and thoracolumbar spine injuries can be managed safely and effectively with thoracic pedicle screws inserted under multiplanar fluoroscopic imaging. PMID- 12544965 TI - Lumbar discography using gadolinium in patients with iodine contrast allergy followed by postdiscography computed tomography scan. AB - STUDY DESIGN: A series of four case reports are presented. OBJECTIVE The case reports document completion of lumbar discography with gadolinium contrast followed by successful computed tomography scanning of intradiscal gadolinium. SUMMARY OF BACKGROUND DATA: Lumbar discography using gadolinium in cases of iodinated contrast allergy with magnetic resonance imaging used to visualize intradiscal gadolinium after the procedure has been described. To the authors' knowledge, there are no reports of gadolinium discography followed by computed tomography scanning of intradiscal gadolinium. METHODS: The reported cases involved a chart review, performance of discography with gadolinium under fluoroscopic guidance, and postdiscography computed tomography scanning of the discs injected during the procedure. RESULTS: Gadolinium was visible under fluoroscopy in all four cases. It was visualized on postdiscography computed tomography scans performed both early and late after the procedure. Normal discs retained gadolinium for a longer period than discs with annular disruption. CONCLUSIONS: Gadolinium discography is a safe, viable alternative to conventional discography in the setting of a patient with iodinated contrast allergy. Postdiscography computed tomography scans adequately visualize intradiscal gadolinium in a more timely and cost-effective manner than magnetic resonance imaging. PMID- 12544966 TI - Localized venous plexi in the spine simulating prolapse of an intervertebral disc: a report of six cases. AB - STUDY DESIGN: Six cases are reported in which the clinical presentation of a prolapsed intervertebral disc was found to be caused by a localized venous plexus. OBJECTIVES: To emphasize the fact that the clinical presentation of a localized plexus of epidural veins in the lumbar spine can resemble that of an acute disc prolapse. SUMMARY OF BACKGROUND DATA: The finding of enlarged epidural veins during lumbar disc decompression is relatively common, but it is only recently that they have been implicated as the cause of the presenting symptoms. METHODS AND RESULTS: Six individuals presented with severe pain in the lower back accompanied by sciatica, which had begun acutely. Physical examination in most of these patients showed the presence of neurologic signs in the affected leg. The initial clinical assessment was that of an acute prolapse of a lumbar disc. This diagnosis appeared to be borne out by the MRI scans, which demonstrated a "prolapsed disc" at the relevant level of the spine. However, at surgery, the intervertebral disc appeared to be relatively normal, but at the spine was found a large, localized plexus of epidural veins whose configuration matched the MRI image. The symptoms were relieved by decompression of the spine and ablation of the veins. CONCLUSIONS: Any pathologic process in the lumbar spine compressing a nerve root can cause localized pain in the back accompanied by sciatica. Most of these conditions can be differentiated by means of an MRI scan. The MRI image of a localized plexus of epidural veins, however, closely resembles that of a prolapsed intervertebral disc, which may be diagnostically misleading. PMID- 12544967 TI - Acute torticollis in an adolescent: case report and MRI study. AB - STUDY DESIGN: A case report is presented. OBJECTIVES: To describe a typical case of acute adolescent torticollis in which a disc lesion was detected with magnetic resonance imaging. SUMMARY OF BACKGROUND DATA: Acute torticollis is attributed to atlantoaxial rotary fixation of unknown etiology. The current view is that the lesion is caused by synovial fold entrapment in the C1-C2 interspace. METHODS: In a 15-year-old male adolescent, magnetic resonance imaging was performed a few hours after the onset of torticollis, and 3 weeks after resolution of symptoms.RESULTS Increased signal intensity compatible with a fluid collection was seen in the right uncovertebral region at C2-C3. This lesion was probably linked to a sudden disruption of the disc collagen fibers, and had caused excessive lateral pressure, pushing C2 toward the left. Magnetic resonance imaging at 3 weeks was unremarkable. CONCLUSIONS: The authors think that the observed disruption was a sudden and abnormal instance of a normal and, usually, very slow process of cleft formation with extension into the fibrocartilaginous core in the uncovertebral region. It is felt that this lesion may be a frequent cause of torticollis in adolescents. PMID- 12544968 TI - Hyperkyphosis as an indicator of syringomyelia in idiopathic scoliosis: a case report. AB - STUDY DESIGN: A case of thoracic hyperkyphosis is reported as an atypical presentation of otherwise typical adolescent idiopathic scoliosis that showed a large syringomyelia on magnetic resonance imaging. OBJECTIVE To review atypical curve patterns that indicate the need for more intensive preoperative workup including a magnetic resonance imaging scan. SUMMARY OF BACKGROUND DATA: The literature on atypical curve patterns of adolescent "non-idiopathic" scoliosis is reviewed. No report in the literature describes patients with an idiopathic coronal plane and thoracic hyperkyphosis as an atypical feature found to have a large syringomyelia and Chiari I malformation on magnetic resonance imaging examination. METHODS: The clinical and radiographic features associated with thoracic hyperkyphosis and juvenile and adolescent scoliosis with syringomyelia are presented. RESULTS: The neurosurgical treatment of the syringomyelia subsequently led to a potentially safer spinal fusion. The patient tolerated the procedures well and at this writing has remained asymptomatic during 3 years of follow-up evaluation. CONCLUSIONS: The reported case adds to the hyperkyphosis literature category of unusual spinal deformities seen in scoliosis associated with syringomyelia and should further help to categorize adolescent idiopathic scoliosis as a diagnosis of exclusion. PMID- 12544969 TI - A third-degree burn caused by a neurogenic motor-evoked potential monitoring electrode during spinal surgery: a case report. AB - STUDY DESIGN: A case report is presented. OBJECTIVE: To report a previously undescribed complication related to use of a neurogenic motor-evoked potential monitoring electrode and electrosurgery during spinal surgery. SUMMARY OF BACKGROUND DATA: Although electrosurgery is one of the most commonly used technologies in the operating room, its electrophysical properties, including the potential for complications, are poorly understood by many surgeons. Complications related to the use of electrosurgical instruments, monitoring electrodes, and radiofrequency current are underreported in the literature. METHODS: Clinical case analysis and investigation report were used. RESULTS: A case of third-degree skin burn at the site of a neurogenic motor-evoked potential monitoring electrode during posterior spinal fusion surgery is described. CONCLUSIONS: A burn resulting from a neurogenic motor-evoked potential monitoring electrode is rare. However, surgeons and electrophysiologists should be familiar with this potential complication in order to prevent it. PMID- 12544971 TI - Economic evaluation of new therapies in critical illness. AB - The recent Food and Drug Administration approval of drotrecogin alfa (activated) and the potential of several other new therapies may represent the beginning of a breakthrough in the management of critical illness in the intensive care unit. However, their use in clinical practice will likely be dependent on a rigorous appraisal not only of their effects, but also of their costs. Novel therapies can no longer be judged simply by their effectiveness in treating illness, but must also be evaluated on an institutional and societal level on the basis of their cost. These considerations have important implications for the practicing intensivist, who will need to better understand the conduct and design of economic evaluations, including their strengths and weaknesses. In this article, we review the rationale behind economic evaluations of new therapies and the alternative economic approaches available. We then discuss in more detail the elements contained in a cost-effectiveness analysis, the preferred approach to pharmacoeconomic evaluation today. PMID- 12544970 TI - Proteomics and systems biology approaches to signal transduction in sepsis. AB - Sepsis and resulting multiple system organ failure are the leading causes of mortality in intensive care units. Although it is generally appreciated that rampant, deregulated inflammatory pathways play a major role in sepsis, a comprehensive understanding based on the integrated response of multiple signal transduction pathways has remained elusive. Here we review the main signal transduction pathways involved in the progression from inflammation to sepsis and discuss emerging genomic, proteomic, and systems biology approaches to decipher how these signaling pathways converge to cause the septic state. We propose that an integrative approach involving functional proteomics will provide a quantitative and mechanistic description that unifies inflammatory signaling networks in sepsis and will identify critical regulatory nodes for therapeutic manipulation. These types of systems biology-based approaches may lead to more effective therapies than those currently available. PMID- 12544972 TI - Impact of intensive care unit (ICU) drug use on hospital costs: a descriptive analysis, with recommendations for optimizing ICU pharmacotherapy. AB - OBJECTIVE: To determine the cost impact of intensive care unit (ICU) drug use on a hospital's total drug expense and to compare ICU pharmacy resource utilization with resource utilization of other hospital departments that provided services to the ICU. Additionally, to suggest strategies, based on these methods and results, to optimize ICU drug use. METHODS: Financial transactions for all ICU patients (n = 23,107) treated during fiscal years 1999-2002 were retrieved from the hospital's data repository. ICU drug costs were calculated both as the percentage of total drug costs for each fiscal year and adjusted for hospital volume (ICU patient days). ICU department charges were calculated as a percentage of total ICU charges and analyzed by simple descriptive statistics (mean +/- sd). Drug utilization was retrieved for those patients accounting for the highest percentage of ICU pharmacy charges. MAIN RESULTS: ICU drug costs accounted for 38.4% (+/-4.1% sd) of the total drug costs and have increased at a rate greater than non-ICU drug costs (12% vs. 6%). ICU pharmacy charges accounted for an average of 11.1% (+/-9.2% sd) of the total ICU charges, ranking as the fourth most costly of ICU charges. Both costly and highly used ICU therapies were identified for focus on cost-effectiveness analyses or application of an evidence based drug use and disease state management program model to ICU pharmacotherapy. CONCLUSIONS: ICU drug therapies have a significant impact on hospital costs, and effective clinical informatics services and multidisciplinary collaboration programs are necessary to optimize ICU pharmacotherapy. PMID- 12544973 TI - Restrictive antibiotic policies are appropriate in intensive care units. AB - Antibiotic use will always be an important part of medical practice in the intensive care unit. Antibiotic resistance increases the chance that empirical therapy will be inadequate to cover the organisms implicated in any particular infection. Therefore, strategies that can allow for optimal empirical antibiotic choice, while at the same time minimizing emergence of antibiotic resistance, are particularly important. In many situations, such strategies require some external stewardship of antibiotic use to be maximally effective. Antibiotic stewardship programs may take the form of management teams comprising infectious disease physicians and pharmacists. These clinicians work in concert with critical care specialists in choosing optimal empirical regimens and in streamlining therapy once culture results are available. Alternatively, computer-based clinical support systems have been developed that can guide physicians to utilize optimal antibiotic choices. External stewardship of antibiotic use may be particularly necessary in circumstances of increased antibiotic resistance, especially exhibited by Gram-negative bacilli. A number of examples exist in which antibiotic control programs can work when traditional infection control programs have failed. Mutation of organisms to produce antibiotic resistance is undoubtedly going to outstrip availability of new antibiotics in the near future. Antibiotic stewardship in concert with improved diagnostic methods may be our only hope in preventing endemic panresistant organisms. PMID- 12544974 TI - Therapeutic potential of superoxide dismutase mimetics as therapeutic agents in critical care medicine. AB - Oxidative stress results from an oxidant/antioxidant imbalance, an excess of oxidants, or a depletion of antioxidants. A considerable body of recent evidence suggests that oxidative stress and exaggerated production of reactive oxygen species play a major role in several aspects of septic shock and ischemia and reperfusion. Initiation of lipid peroxidation, direct inhibition of mitochondrial respiratory chain enzymes, inactivation of glyceraldehyde-3-phosphate dehydrogenase, inhibition of membrane Na /K adenosine triphosphatase activity, inactivation of membrane sodium channels, and other oxidative protein modifications contribute to the cytotoxic effect of reactive oxygen species. In addition, reactive oxygen species are potent triggers of DNA strand breakage, with subsequent activation of the nuclear enzyme poly-adenosine 5'-diphosphate ribosyl synthetase, and eventual severe energy depletion of the cells. Pharmacologic evidence suggests that the peroxynitrite-poly-adenosine 5' diphosphate ribosyl polymerase pathway contributes to the cellular injury in shock and endothelial injury. Treatment with superoxide dismutase mimetics, which selectively mimic the catalytic activity of the human superoxide dismutase enzymes, has been shown to prevent the cellular energetic failure associated with shock and ischemia-reperfusion and to prevent tissue damage associated with these conditions. In this article, we will briefly review the role of superoxide in septic shock and ischemia-reperfusion injury. We hope to present evidence to support the potential development of superoxide dismutase mimetics as novel and effective agents in the area of critical care medicine. PMID- 12544975 TI - Role of flagellin in the pathogenesis of shock and acute respiratory distress syndrome: therapeutic opportunities. AB - OBJECTIVE: To provide an overview of the role of flagellin as an immunostimulatory and proinflammatory factor. DESIGN: A systematic overview of the literature on this subject. SETTING: An office equipped with a computer and Internet access to PubMed. SUBJECTS: MEDLINE citations between 1960 and 2002. MAIN RESULTS: Flagellin, a protein of 40-60 kD, is the principal constituent of the flagellum, a prominent surface structure found in motile bacteria. Recent work reveals that monomeric flagellin, a protein component of flagellated bacteria, can act as a soluble immunostimulatory and proinflammatory factor, activating the immune/inflammatory axis via the toll-like receptor 5-nuclear factor-kappaB axis. Monocytes, macrophages, and intestinal and pulmonary epithelial cells respond to monomeric flagellin at low concentrations. Monomeric flagellin can induce prominent local and systemic immune/inflammatory responses and. CONCLUSIONS: Recognition of the flagellin-toll-like receptor 5 pathway offers novel opportunities for the experimental therapy of various forms of shock, sepsis, and acute respiratory distress syndrome. PMID- 12544976 TI - Targeting high mobility group box 1 as a late-acting mediator of inflammation. AB - Sepsis, a lethal systemic inflammatory response to infection, affects nearly 750,000 patients in the United States annually and has a mortality of 30%. Mounting evidence has implicated cytokines, circulating factors produced by the innate immune system, as critical mediators of sepsis-related tissue injury and death. Many resources have been expended to elucidate the pathologic mechanisms that underlie sepsis and to develop appropriate and effective therapeutics. To date, no anti-inflammatory agent has been clinically approved for the treatment of sepsis because even a slight delay in administration of therapeutics that target inflammatory mediators renders most approaches ineffective. These and other findings, described in part in this review, suggest that successful clinical management of sepsis may be dependent on identification of late-acting, downstream lethal mediators that can be targeted in a broader therapeutic window. A candidate mediator of delayed lethality is high mobility group box 1, a cellular and nuclear protein that is now recognized as a cytokine and experimental therapeutic target. PMID- 12544977 TI - Ethyl pyruvate: a novel anti-inflammatory agent. AB - Pyruvate plays a central role in intermediary metabolism. Pyruvate, however, is also a potent antioxidant and free radical scavenger, and numerous studies have shown that treatment with this compound can be salutary in numerous pathologic conditions that are thought to be mediated, at least in part, by redox-dependent phenomena. Unfortunately, aqueous solutions of pyruvate rapidly undergo an aldol like condensation reaction to form 2-hydroxy-2-methyl-4-ketoglutarate (parapyruvate), a compound that is a potent inhibitor of a critical step in the mitochondrial tricarboxylic acid cycle. To circumvent this issue, our laboratory formulated a derivative of pyruvic acid, ethyl pyruvate, in a calcium- and potassium-containing balanced salt solution. We showed that treatment with this fluid could ameliorate much of the structural and functional damage to the intestinal mucosa caused by mesenteric ischemia and reperfusion in rats. In subsequent studies, we showed that treatment with ethyl pyruvate solution could improve survival in rodent models of hemorrhagic shock and resuscitation and also down-regulate a number of proinflammatory genes. Recently, ethyl pyruvate was also shown to improve survival in murine models of acute endotoxemia and bacterial peritonitis. Although the biochemical basis for the anti-inflammatory actions of pyruvate remain to be elucidated, this simple compound warrants further evaluation as a treatment for a number of conditions commonly encountered in the practice of critical care medicine. PMID- 12544978 TI - Endotoxin as a drug target. AB - OBJECTIVE: To review the preclinical and clinical evidence that antiendotoxin therapeutic strategies are potentially useful in the prevention and treatment of septic shock. STUDY DESIGN: A critical review of the literature over the past 30 yrs relating basic and clinical research on the therapeutic value of endotoxin as a target for the prevention and treatment of severe sepsis and septic shock. MAIN RESULTS: Bacterial endotoxin is a potent and predominant microbial mediator that induces an intense inflammatory and procoagulant response by elements of the innate immune response. This macromolecule is capable of inducing lethal septic shock in experimental animals, and a large number of preclinical studies consistently demonstrate the survival advantage of endotoxin inhibition in experimental models of sepsis. Clinical studies indicate that endotoxin may be found in the systemic circulation in the majority of humans with septic shock. Endotoxemia is largely independent of the nature of the infecting microorganism despite the fact that this molecule is specifically found in the outer membrane of Gram-negative bacteria only. Antiendotoxin strategies studied thus far have not provided reproducible survival benefits in clinical trials in septic patients. CONCLUSIONS: Despite compelling evidence of the critical importance of endotoxin in the pathogenesis of Gram-negative bacterial sepsis in preclinical investigations and numerous clinical interventional trials, the utility of antiendotoxin approaches to significantly reduce the mortality rate in human septic shock remains unproven. Ongoing clinical trials with specific endotoxin inhibitors should determine the potential value of this therapeutic approach to the management of septic shock. PMID- 12544979 TI - How to leverage an endogenous immune defense mechanism: the example of granulocyte colony-stimulating factor. AB - Our understanding of host defense has exploded during the past two decades. It is temping to take advantage of this knowledge by considering the modulation and control of these mechanisms as therapeutic options. In intensive care medicine, the aim is usually to block an overwhelming inflammatory response, which represents the "bad" side of the double-edged sword of host defense. The obvious danger of such treatment strategies is that impairing the inflammatory reaction means impairing host defense in patients exposed to infectious agents. The alternative approach, i.e., strengthening or supplementing favorable host defense mechanism, has so far been little explored clinically. Granulocyte colony stimulating factor, the granulocyte colony-stimulating factor, combines the unique properties of an anti-infectious and an anti-inflammatory factor. This attractive profile has led us to various approaches to exploit these immunomodulatory activities. In a recently terminated, placebo-controlled, randomized study, we investigated if prophylactic treatment with rh granulocyte colony-stimulating factor (Filgrastim), at the time a risk can be anticipated such as before an operation, may offer protection from immunoinflammatory dyshomeostasis and thus lower the incidence of postoperative sepsis. Perioperative rh granulocyte colony-stimulating factor administration, compared with placebo treatment, resulted in the prevention of postoperative monocyte deactivation, conservation of an adequate Th1/Th2 ratio, as well as a considerable alleviation of the acute phase response. In parallel, there was a clear tendency toward lowering the rate of postoperative septic complications under the administration of Filgrastim. PMID- 12544980 TI - Membrane-permeable radical scavengers (tempol) for shock, ischemia-reperfusion injury, and inflammation. PMID- 12544981 TI - Drotrecogin alfa (activated) (recombinant human activated protein C) for the treatment of severe sepsis. AB - OBJECTIVE: To review the data supporting drotrecogin alfa (activated) for severe sepsis treatment. DATA SOURCES: Published research and data from the Protein C Worldwide Evaluation in Severe Sepsis (PROWESS) trial. DATA EXTRACTION AND SYNTHESIS: The coagulation cascade and intense inflammation play a central role in the development of organ failure due to severe sepsis. Drotrecogin alfa (activated) has anti-inflammatory, antithrombotic, profibrinolytic, and other properties that may explain the beneficial results seen in both animal models and humans with severe sepsis. Drotrecogin alfa (activated) produces a robust reduction in the mortality rate of patients with severe sepsis that is evident across nearly every subgroup examined in the phase III clinical trial and has an acceptable safety profile with bleeding during infusion as the only significant risk associated with therapy. The relative risk reductions for mortality seen in Gram-negative, Gram-positive, pneumonia, abdominal sources, shock, and nonshock are similar to the intent-to-treat population, 19.4%. Treatment also increases days alive and free from mechanical ventilation and shock. CONCLUSIONS: Coagulopathy and systemic inflammation are almost universal in patients with severe sepsis. Treatment of this disorder with drotrecogin alfa (activated) directly addresses these derangements and substantially reduces morbidity and mortality rates with potential for bleeding during infusion as the only known risk. PMID- 12544982 TI - Recombinant human activated protein C in sepsis: inconsistent trial results, an unclear mechanism of action, and safety concerns resulted in labeling restrictions and the need for phase IV trials. PMID- 12544983 TI - Therapeutic potential of targeting the complement cascade in critical care medicine. AB - Caring for the critical care patient involves many different areas of clinical expertise and serves a diverse patient population. Novel therapeutics for the critically ill must be approached with caution, because the underlying molecular mechanisms of the disease process for several commonly seen types of patients (i.e., sepsis, shock, ischemia/reperfusion) are not fully understood. A potentially new and advancing area of therapeutics that may hold promise for the critically ill is inhibition of the complement system. Various novel complement inhibitors are being developed and several are in clinical trials. The advancement of this novel area of therapeutics may one day aid the clinician by providing several different complement inhibitors/antagonists for controlling complement activation or its biologically active mediators. PMID- 12544984 TI - Nuclear factor-kappaB as a therapeutic target in critical care medicine. AB - Nuclear factor-kappaB is a transcriptional factor required for the gene expression of many inflammatory mediators. Nuclear factor-kappaB activation requires removal and degradation of its inhibitor kappaB, an event that occurs after phosphorylation of inhibitor kappaB by a complex of inhibitor kappaB kinases. These events allow nuclear factor-kappaB to translocate into the nucleus, where it binds to kappaB elements and initiates transcription. Inappropriate and prolonged activation of nuclear factor-kappaB has been linked to several diseases associated with inflammatory events, including septic shock, acute respiratory distress syndrome, ischemia, and reperfusion injury. Thus, the key role of nuclear factor-kappaB in regulating inflammation makes this factor a therapeutic target for reducing tissue and organ damage. Regulation and control of nuclear factor-kappaB can be achieved by gene modification strategies or by pharmacologic inhibition of the key components of the cascade that leads to nuclear factor-kappaB activation. The purpose of our review is to describe these novel therapeutic approaches and their potential efficacy. PMID- 12544985 TI - Promising strategies to minimize secondary brain injury after head trauma. AB - OBJECTIVE: To review novel therapeutic approaches in the treatment of severe traumatic brain injury. DESIGN: Eighty-three studies were reviewed specific to the treatment of traumatic brain injury, in either experimental models or in patients. CONCLUSION: Four therapeutic strategies appear to be the most promising approaches currently in clinical trials for severe traumatic brain injury: a) the novel pharmacologic agent dexanabinol; b) hypertonic saline; c) mild hypothermia; and d) decompressive craniectomy. Each of these therapies share the common feature of targeting multiple mechanisms, suggesting this may be an important factor to the development of a successful approach to severe traumatic brain injury. PMID- 12544986 TI - Cost-effectiveness of drotrecogin alfa (activated) in the treatment of severe sepsis. AB - OBJECTIVE: To assess the cost-effectiveness of drotrecogin alfa (activated) therapy, which was recently shown to reduce mortality in severe sepsis. DESIGN: Estimates of effectiveness and resource use were based on data collected prospectively as part of a multicenter international trial. Estimates of hospital costs were based on a subset of the patients treated in the United States (33% of all enrolled patients). Lifetime projections were modeled from published sources and tested in sensitivity analyses. Analyses were conducted from the United States societal perspective, limited to healthcare costs, and using a 3% annual discount rate. SETTING: A total of 164 medical institutions in 11 countries. PATIENTS: Adults > or = 18 yrs of age with severe sepsis INTERVENTIONS: Eligible patients were randomly assigned to receive a 96-hr intravenous infusion of drotrecogin alfa (activated) at 24 microg/kg/hr (n = 850) or placebo (n = 840). MEASUREMENTS AND MAIN RESULTS: Base Case: incremental short-term (days 1-28) healthcare costs per day-28 survivor; Panel on Cost-Effectiveness in Health and Medicine Reference Case: incremental lifetime healthcare costs per quality adjusted life-year. Over the first 28 days (short-term Base Case), drotrecogin alfa (activated) increased the costs of care by $9,800 and survival by 0.061 lives saved per treated patient. Thus, drotrecogin alfa (activated) cost $160,000 per life saved (with 84.7% probability that ratio is <$250,000 per life saved). Projected to lifetime (lifetime Reference Case), drotrecogin alfa (activated) increased the costs of care by $16,000 and quality-adjusted survival by 0.33 quality-adjusted life-years per treated patient. Thus, drotrecogin alfa (activated) cost $48,800 per quality-adjusted life-year (with 82% probability that ratio is <$100,000 per quality-adjusted life-year). Estimates were generally robust to sensitivity analyses, although cost-effectiveness deteriorated to >$100,000 per quality-adjusted life-year if survivors lived <4.6 yrs on average. Drotrecogin alfa (activated) cost $27,400 per quality-adjusted life-year when limited to patients with an Acute Physiology and Chronic Health Evaluation II score > or = 25 and was cost-ineffective when limited to patients with a score <25. CONCLUSIONS: Drotrecogin alfa has a cost-effectiveness profile similar to that of many well-accepted healthcare strategies and below commonly quoted thresholds. PMID- 12544987 TI - Drotrecogin alfa (activated) administration across clinically important subgroups of patients with severe sepsis. AB - OBJECTIVE: To assess the effects of drotrecogin alfa (activated) therapy, a recombinant human activated protein C, across clinically relevant subpopulations in a randomized, phase 3, placebo-controlled study of patients with severe sepsis (recombinant human activated protein C worldwide evaluation in severe sepsis [PROWESS]). DESIGN: Univariate and multivariable analysis of prospectively defined subgroups from the PROWESS study. SETTING: A total of 164 medical centers in 11 countries. PATIENTS: A total of 1,690 patients with severe sepsis. MEASUREMENTS AND MAIN RESULTS: We report observed 28-day mortality rates for drotrecogin alfa (activated) and placebo patients for subgroups prospectively defined by demographic data, surgical status, type and site of infection, and clinical and biochemical measures of disease severity. We performed subgroup analyses to explore the consistency of the mortality benefit observed in the overall population and performed tests for both quantitative and qualitative interactions. To examine the magnitude of the treatment benefit with drotrecogin alfa (activated) across the underlying predicted risk of mortality spectrum, we used stepwise logistic regression on PROWESS placebo patients to generate a predicted risk of mortality model that simultaneously included many clinical and biochemical markers of mortality risk. Because drotrecogin alfa (activated) has anticoagulant properties, we also present analyses of bleeding and thrombotic events. Actual mortality rates were lower with drotrecogin alfa (activated) compared with placebo for nearly all prospectively defined subgroups. Both univariate and multivariable regression analyses showed a consistent relative risk reduction in 28-day mortality rates for drotrecogin alfa (activated). Larger absolute risk reductions were found with drotrecogin alfa (activated) in patients with a higher baseline predicted risk of mortality, and actual mortality rates were lower with drotrecogin alfa (activated) in all subgroups defined by disease severity measures where a > or = 20% placebo mortality was observed. Although discriminatory power was limited by few observed events, the increased absolute risk of experiencing a serious bleeding event with treatment did not seem to vary according to the baseline predicted risk of mortality. CONCLUSIONS: The administration of drotrecogin alfa (activated) to patients with severe sepsis was associated with a significant survival benefit that tended to increase with higher baseline likelihood of death. Current data suggest that the increased risk of bleeding does not vary according to likelihood of death. PMID- 12544988 TI - Prognostic value of surfactant proteins A and D in patients with acute lung injury. AB - OBJECTIVE: The primary objective of this study was to test the hypothesis that in patients intubated for acute lung injury, lower concentrations of surfactant proteins A and D in the pulmonary edema fluid and higher concentrations in the plasma are associated with more severe lung injury and worse clinical outcomes. DESIGN: Observational study. SETTING: Intensive care unit patients in a tertiary university hospital and a university-affiliated city hospital. PATIENTS: Thirty eight intubated, mechanically ventilated intensive care unit patients with acute lung injury or acute respiratory distress syndrome as defined by the North American European Consensus Conference. INTERVENTIONS: Undiluted pulmonary edema fluid and plasma samples were collected within 24 hrs of endotracheal intubation in all patients. MEASUREMENTS AND MAIN RESULTS: The concentrations of surfactant proteins A and D were measured in pulmonary edema fluid and in plasma. Plasma surfactant protein A, but not surfactant protein D, was higher in patients with fewer days of unassisted ventilation (p = .03) and in patients with an absence of intact alveolar fluid clearance (p =.03). In contrast, pulmonary edema fluid surfactant protein D, but not surfactant protein A, was lower in patients with worse oxygenation, as measured by the alveolar-arterial oxygen difference (p = .01) and was lower in the patients who died (2646 ng/mL) compared with those who survived (5503 ng/mL; p = .02). CONCLUSIONS: These results demonstrate that reduced pulmonary edema fluid surfactant protein D and elevated plasma surfactant protein A concentrations at the onset of acute lung injury may be associated with more severe disease and worse clinical outcome and may serve as valuable biochemical markers of prognosis. PMID- 12544989 TI - Risk factors for long intensive care unit stay after cardiopulmonary bypass in children. AB - OBJECTIVES: To determine whether children who experience longer intensive care unit (ICU) stays after open heart surgery may be identified at admission by clinical criteria. To identify factors associated with longer ICU stays that are potential targets for quality improvement. SETTING: Tertiary pediatric cardiac surgical center. DESIGN: A retrospective review was performed of pre-, intra-, and postoperative factors for children undergoing open heart surgery. All factors were evaluated for strength of association with length of ICU stay (LOS) using a negative binomial model. After multiple analysis, factors were deemed significant if associated with a LOS with p < .02. PATIENTS: A total of 355 pediatric patients who had cardiac surgery with cardiopulmonary bypass in a 1-yr period from April 1999 until March 2000. MEASUREMENTS AND MAIN RESULTS: Children who fell above the 95th percentile for LOS in our institution occupied 30% of bed days and had a three-fold greater mortality. Of all clinical factors considered, those significantly associated with LOS were as follows: preoperative--mechanical ventilation, neonatal status, medical problems, and transfer from abroad; intraoperative--higher operative complexity, increased cardiopulmonary bypass time or ischemic time, and circulatory arrest; and postoperative--delayed sternal closure, sepsis, renal failure, pulmonary hypertension, chylothorax, diaphragm paresis, and arrhythmia. A model combining all factors identified preoperative mechanical ventilation, neonatal status, major medical problems, operative complexity, cardiopulmonary bypass time, and a postoperative complication score as independently associated with LOS (p < .01). CONCLUSIONS: At the time of ICU admission after open heart surgery, clinical criteria are evident that highlight a child's risk of longer ICU stay. These pre- and intraoperative factors relate to LOS independent of subsequent postoperative events. Those postoperative complications that are most strongly associated with increased LOS are identified and, therefore, made accessible to quality control. PMID- 12544990 TI - Influence of interleukin-10 polymorphisms on interleukin-10 expression and survival in critically ill patients. AB - OBJECTIVE: To determine the functionality of identified polymorphisms in the promoter and upstream regions of the interleukin-10 gene in terms of release of interleukin-10 from lipopolysaccharide-stimulated whole blood from healthy volunteers and to evaluate the relationship of interleukin-10 polymorphisms to interleukin-10 release, development of sepsis, and mortality in critically ill patients. DESIGN: Observational study. SETTING: The academic unit of anesthesia and intensive care, university laboratories, and ten-bed general intensive care unit in a university teaching hospital. SUBJECTS: A total of 132 healthy volunteers plus 67 consecutive critically ill patients recruited within 24 hrs of admission to the intensive care unit, regardless of diagnosis. MEASUREMENTS: Plasma interleukin-10 levels were measured by enzyme-linked immunosorbent assay. Single nucleotide polymorphisms were detected by restriction fragment length polymorphism analysis. Dinucleotide repeat polymorphisms were identified after polymerase chain reaction using a DNA size analyzer. MAIN RESULTS: Stimulated interleukin-10 release in critically ill patients was significantly lower than in healthy subjects (p < .0001). In addition, in the patients who developed sepsis, interleukin-10 release at admission to the intensive care unit was significantly lower than in patients who did not subsequently develop sepsis (median [range] 1.47 [0.13-6.90] ng/mL compared with 4.93 [0.03-16.80] ng/mL, p = .001). The A allele of the single nucleotide polymorphism at -592 base pairs was associated with lower interleukin-10 release and higher mortality in critically ill patients. Other polymorphisms were not linked to interleukin-10 release, sepsis, or mortality. CONCLUSIONS: The A allele of the -592 base pair single nucleotide polymorphism in the interleukin-10 gene is associated with lower stimulated interleukin-10 release and increased mortality. Further investigations are required to determine the nature of the functionality and the potential diagnostic and therapeutic aspects of this marker. PMID- 12544991 TI - Intravenous erythromycin facilitates bedside placement of postpyloric feeding tubes in critically ill adults: a double-blind, randomized, placebo-controlled study. AB - OBJECTIVE To evaluate the efficacy of intravenous erythromycin as a method to facilitate feeding tube placement into the small intestine in critically ill patients. DESIGN Double blind, randomized, controlled trial. SETTING Medical and surgical intensive care units in an academic medical center. PATIENTS Prospective cohort of 36 consecutive adults requiring intensive care unit care and enteral tube feeding for nutritional support. INTERVENTION Infusion of a single dose of intravenous erythromycin (500 mg) or saline before placement of 10-Fr feeding tubes using a standardized active bedside protocol. MEASUREMENTS AND MAIN RESULTS We determined the success rate of feeding tube placement into or beyond the second portion of the duodenum and the time required for this procedure by experienced nurses. The feeding tube was considered to be postpyloric when the tip was in the second portion of the duodenum or beyond. The predictive value of a serial step-up in gastrointestinal aspirate pH from < or = 5.0 to > or = 6.0 was also determined. Use of intravenous erythromycin significantly improved the rate of feeding tube placement into the duodenum or jejunum (erythromycin group, 13 of 14 patients or 93% vs. the control group, 12 of 22 patients or 55%; p < .03). Erythromycin administration also significantly decreased the procedure time from 25 +/- 3 to 15 +/- 2 mins (p < .04). Feeding tube placement into either duodenum or jejunum was confirmed in all 18 patients with a pH step-up from < or = 5.0 to > or = 6.0. CONCLUSION: A single bolus dose of intravenous erythromycin facilitates active bedside placement of postpyloric feeding tubes in critically ill adult patients. PMID- 12544992 TI - Early indicators of prolonged intensive care unit stay: impact of illness severity, physician staffing, and pre-intensive care unit length of stay. AB - OBJECTIVE: Scoring systems that predict mortality do not necessarily predict prolonged length of stay or costs in the intensive care unit (ICU). Knowledge of characteristics predicting prolonged ICU stay would be helpful, particularly if some factors could be modified. Such factors might include process of care, including active involvement of full-time ICU physicians and length of hospital stay before ICU admission. DESIGN: Demographic data, clinical diagnosis at ICU admission, Simplified Acute Physiology Score, and organizational characteristics were examined by logistic regression for their effect on ICU and hospital length of stay and weighted hospital days (WHD), a proxy for high cost of care. SETTING: A total of 34 ICUs at 27 hospitals participating in Project IMPACT during 1998. PATIENTS: A total of 10,900 critically ill medical, surgical, and trauma patients qualifying for Simplified Acute Physiology Score assessment. INTERVENTIONS: None. RESULTS: Overall, 9.8% of patients had excess WHD, but the percentage varied by diagnosis. Factors predicting high WHD include Simplified Acute Physiology Score survival probability, age of 40 to 80 yrs, presence of infection or mechanical ventilation 24 hrs after admission, male sex, emergency surgery, trauma, presence of critical care fellows, and prolonged pre-ICU hospital stay. Mechanical ventilation at 24 hrs predicts high WHD across diagnostic categories, with a relative risk of between 2.4 and 12.9. Factors protecting against high WHD include do-not-resuscitate order at admission, presence of coma 24 hrs after admission, and active involvement of full-time ICU physicians. CONCLUSIONS: Patients with high WHD, and thus high costs, can be identified early. Severity of illness only partially explains high WHD. Age is less important as a predictor of high WHD than presence of infection or ventilator dependency at 24 hrs. Both long ward stays before ICU admission and lack of full-time ICU physician involvement in care increase the probability of long ICU stays. These latter two factors are potentially modifiable and deserve prospective study. PMID- 12544993 TI - Impact of oligon central venous catheters on catheter colonization and catheter related bloodstream infection. AB - OBJECTIVE: To evaluate a new antimicrobial treatment for central venous catheters in comparison with a traditional treatment, by assessing the catheter colonization and catheter-related bloodstream infection rates in two groups of patients. DESIGN: Multiple-center, prospective randomized study. SETTING: The medical and surgical departments of ten institutions. PATIENTS: Patients requiring a central venous catheter for medical or surgical pathologies between June 2000 and November 2001. INTERVENTIONS: Patients in the control group received a conventional benzalkonium-treated double-lumen central venous catheter, while patients in the oligon group received an oligon-treated (polyurethane combined with silver, carbon, and platinum) catheter with the same characteristics. Data collection included demographics, preexisting clinical conditions, main pathology, catheter insertion, and management data. Catheter colonization was defined as the growth of > or = 15 colony-forming units in culture of catheter segments by the roll-plate method, or > or = 1000 colony forming units for the sonication method, and catheter-related bloodstream infection was defined as isolation of the same organism from the colonized catheter and from the peripheral blood of a patient with clinical signs of bloodstream infection. MEASUREMENTS AND MAIN RESULTS: Data were obtained from 545 catheters. Of these, 132 catheters (24.2%) were positive for colonization. Patients in the oligon group demonstrated a lower risk for catheter colonization in the overall population (relative risk, 0.63; 95% confidence interval, 0.46 0.86; p = .003) and in the surgical subgroup (relative risk, 0.5; 95% confidence interval, 0.33-0.76;p = .001). Significant differences between groups were detected for coagulase-negative staphylococci and Gram-negative bacilli colonization rates. Twenty-one patients (3.8%) were positive for catheter-related bloodstream infection, without significant differences between control and oligon groups. CONCLUSIONS: Oligon treatment is effective in limiting the catheter colonization rate. Due to the limited amount of events, this study lacked the power to detect significant differences in terms of catheter-related bloodstream infection rate. PMID- 12544994 TI - Life-sustaining treatments in patients who died of chronic congestive heart failure compared with metastatic cancer. AB - INTRODUCTION: Life-sustaining treatments such as cardiopulmonary resuscitation, mechanical ventilation, vasopressors, and admission to critical care units, if used when recovery chance was remote, may unnecessarily cause discomfort and increase cost of care. Outcomes of these treatments in chronic, refractory congestive heart failure (CHF) and metastatic cancer patients were poor. Although both conditions were the leading causes of death, previous studies indicated that hospice utilization and do-not-resuscitate orders were less common in CHF patients. To date, the use of life-sustaining treatments in these patients and the influence of do-not-resuscitate orders remains unknown. METHOD: We conducted a retrospective medical record review of the patients who died in our hospital in 1999 and had discharge diagnoses of CHF or cancer. Medical records were screened for seriously ill patients according to the modified SUPPORT criteria, which included patients with CHF functional class IV or ejection fraction of 20% or less at baseline and with metastatic cancer not receiving any curative treatments. Analyses were performed using SPSS, version 9.0. RESULTS: There were 58 and 82 patients in CHF and cancer groups, respectively. CHF patients were older (78.8 vs. 67.3 yrs, p < .001) and stayed in the hospital longer (11.9 vs. 7.9 days, p = .014). The majority of patients in both groups received do-not resuscitate orders before death (84% and 72%, respectively). CHF patients received do-not-resuscitate orders later than did cancer patients (6.7 vs. 2.8 days, p = .006). However, there was no significant difference in prevalence of do not-resuscitate orders. All studied life-sustaining treatments were more common in CHF patients than in cancer patients. A subgroup analysis between CHF patients with do-not-resuscitate orders and those without do-not-resuscitate orders revealed cardiopulmonary resuscitation to be the only treatment less common in those with do-not-resuscitate orders. CONCLUSIONS: Patients who died of chronic, refractory CHF received more life-sustaining treatments than did patients who died of metastatic cancer. PMID- 12544995 TI - High and low response in relation to nitric oxide formation but not to lipid peroxidation in patients with sepsis. AB - OBJECTIVES: Nitric oxide overproduction in sepsis syndrome was suspected to be responsible for hemodynamic derangement and, by induction of lipid peroxidation, for tissue damage. Therefore, nitric oxide formation and lipid peroxidation were quantified in septic patients (SP) vs. patients with localized infection (IF) or without inflammation (C). Nitric oxide formation in sepsis was additionally compared with data for clinical status. DESIGN: Prospective study with consecutive sampling of patients. SETTING: A university hospital intensive care unit and research laboratories. PATIENTS: SP, 24 patients; IP, 7; and C, 13. INTERVENTIONS: Plasma measurement of nitrate, lipid peroxides (primary endpoints), and N-hydroxy-L-arginine (secondary end point) MEASUREMENTS AND MAIN RESULTS: For nitrate, there was a sequence of C < IP = SP. Among SP, one group with significantly higher nitrate (high-responders for nitric oxide; SP-HR) vs. IP and C and a second group (low-responders; SP-LR) with increased concentration only vs. C could be identified. For SP-HR vs. IP, a strong time kinetics in nitric oxide formation was obvious, indicated by significant nitrate increase already 1 day before sepsis started, tripling up to the peak concentration, and then a lowering but still increased value on the first day after sepsis. N hydroxy-L-arginine was significantly increased in SP-HR vs. C. For lipid peroxides, the concentrations were comparable in SP and IP, but both significantly increased vs. C. Clustering and coincident kinetics of lipid peroxidation related to nitric oxide were not obvious. Furthermore, there was no strong correlation of clinical data and nitric oxide clustering in sepsis. CONCLUSIONS: High- and low-responders for nitric oxide were identified among septic patients. This finding was not associated with significant differences in lipid peroxidation or clinical data. PMID- 12544997 TI - Femoral central venous catheter-associated deep venous thrombosis in children with diabetic ketoacidosis. AB - OBJECTIVE: To describe the incidence of clinical deep venous thrombosis associated with femoral central venous catheters (CVC-DVT) in children with diabetic ketoacidosis (DKA). DESIGN: Retrospective case-matched control series. SETTING: Pediatric intensive care units of two university-affiliated hospitals. PATIENTS: All eight pediatric DKA patients with femoral central venous catheters between 1998 and 2001, and 16 age-matched control patients with femoral central venous catheters and circulatory shock. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The records of all children with DKA and the control patients were reviewed. CVC-DVT was defined as persistent ipsilateral leg swelling after removal of a femoral central venous catheter. Control patients with coagulopathies, thrombocytopenia, cancer, and hyperglycemia were excluded. Four of eight patients with DKA developed CVC-DVT compared with none of the 16 control patients (p = .007, Fisher's exact test). All four patients with DKA and CVC-DVT were <3 yrs old. Doppler ultrasound examination was performed on three of the four patients with clinical CVC-DVT, confirming the diagnosis in each case. CONCLUSIONS: This study suggests that young children with DKA have an increased incidence of clinical DVT associated with the placement of femoral central venous catheters. PMID- 12544996 TI - Heat shock protein 70 genotypes HSPA1B and HSPA1L influence cytokine concentrations and interfere with outcome after major injury. AB - OBJECTIVE: To examine the influence of genetic variations in heat shock proteins on trauma outcome. DESIGN: Prospective, noninterventional, single-center study. SETTING: Level I trauma center. SUBJECTS: Eighty consecutive severe multiple trauma patients. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Plasma concentrations of interleukin-6 and tumor necrosis factor-alpha were measured over a 5-day course by chemiluminescence-immunoassay. The genotypes of the polymorphisms HSPA1B (HSP70-2) G1538A and HSPA1L (HSP70-Hom) C2437T were determined by polymerase chain reaction and restriction cleavage with PstlI or NcoI, respectively. Allele frequency of the HSPA1B 1538 G allele was 0.569, and that of the HSPA1L 2437 T allele was 0.821. Interleukin-6 concentrations rapidly increased and dropped to almost normal after 5 days, whereas tumor necrosis factor-alpha concentrations increased until day 5. Patients carrying the genotypes HSPA1B AG or HSPA1L CT had significantly higher plasma concentrations of tumor necrosis factor-alpha and interleukin-6 compared with those with genotype GG or TT. Presence of the HSPA1L genotype CT also was a significant risk factor to develop liver failure (odds ratio, 4.6; 95% confidence interval, 1.5 14.1) and to acquire at least one complication severe enough to score three points according to the Denver multiple organ failure score (odds ratio, 3.0; 95% confidence interval, 1.1-9.2). CONCLUSION: The data indicate that genetic variations of the heat shock proteins HSPA1B and HSPA1L may contribute to clinical outcome after severe injury. PMID- 12544998 TI - Right-to-left shunt and risk of decompression illness with cochleovestibular and cerebral symptoms in divers: case control study in 101 consecutive dive accidents. AB - OBJECTIVE: We investigated the role of right-to-left shunt with standardized transcranial Doppler ultrasonography in a large population of divers referred for symptoms of decompression illness. DESIGN: Case series compared with a control group. SETTING: Military teaching hospital, hyperbaric unit. PATIENTS: Patients were 101 consecutive divers with clinical evidence of decompression illness and a control group of 101 healthy divers. INTERVENTION: Specification of the type of decompression illness involved and detection/evaluation of right-to-left shunt by standardized transcranial Doppler. The degree of right-to-left shunt was defined as major if the number of high-intensity transient signals in the middle cerebral artery was >20. MEASUREMENTS AND MAIN RESULTS: We evaluated the odds ratios by logistic regression analysis with vs. without right-to-left shunt for subjects with cochleovestibular symptoms, cerebral decompression illness, spinal decompression illness, and Caisson sickness. Of the 101 divers presenting with decompression illness, transcranial Doppler detected a right-to-left shunt in 59 (58.4%), whereas control subjects demonstrated a right-to-left shunt in 25 cases (24.8%; odds ratio, 4.3; 95% confidence interval, 2.3-7.8; p=.09). When a right to-left shunt was detected, the right-to-left shunt was major in 12 of 25 patients in the control group and in 49 of 59 patients in the decompression illness group (odds ratio, 8.7; 95% confidence interval, 4.2-18.0; p<.001). Within the decompression illness group, the proportion of major right-to-left shunt was 24 of 34 (odds ratio, 29.7; 95% confidence interval, 10.0-87.2; p<.0001) in the cochleovestibular subgroup, 13 of 21 (odds ratio, 24.1, 95% confidence interval, 6.8-86.0, p< 0.0001) in the cerebral decompression illness subgroup, ten of 31 (odds ratio, 3.9; 95% confidence interval, 1.5-10.3; p<.01) in the spinal decompression illness subgroup, and two of two (odds ratio, 1.1; 95% confidence interval, 0.2-5.7; p=.9) in the subgroup of divers with Caisson sickness. CONCLUSION: Based on our results, we conclude that major right-to-left shunt was associated with an increased incidence of cochleovestibular and cerebral decompression illness, suggesting paradoxical embolism as a potential mechanism. PMID- 12544999 TI - Respective effects of end-expiratory and end-inspiratory pressures on alveolar recruitment in acute lung injury. AB - OBJECTIVE: A low tidal volume can induce alveolar derecruitment in patients with acute lung injury. This study was undertaken to evaluate whether this resulted mainly from the decrease in tidal volume or from the reduction in end-inspiratory plateau pressure and whether there is any benefit in raising the level of positive end-expiratory pressure (PEEP) while plateau pressure is kept constant. DESIGN: Prospective crossover study. SETTING: Medical intensive care unit of a university teaching hospital. PATIENTS: Fifteen adult patients ventilated for acute lung injury (PaO2/FiO2, 158 +/- 34 mm Hg; lung injury score, 2.7 +/- 0.6). INTERVENTIONS: Three combinations were tested: PEEP at the lower inflection point with 6 mL/kg tidal volume, PEEP at the lower inflection point with 10 mL/kg tidal volume, and high PEEP with tidal volume at 6 mL/kg, keeping the plateau pressure similar to the preceding condition. MEASUREMENTS AND MAIN RESULTS: Pressure volume curves at zero PEEP and at set PEEP were recorded, and recruitment was calculated as the volume difference between both curves for pressures ranging from 15 to 30 cm H2O. Arterial blood gases were measured for all patients. For a similar PEEP at the lower inflection point (10 +/- 3 cm H2O), tidal volume reduction (10 to 6 mL/kg) led to a significant derecruitment. A low tidal volume (6 mL/kg) with high PEEP (14 +/- 3 cm H2O), however, induced a significantly greater recruitment and a higher Pao than the two other strategies. CONCLUSION: At a given plateau pressure (i.e., similar end-inspiratory distension), lowering tidal volume and increasing PEEP increase recruitment and PaO2. PMID- 12545000 TI - Overestimation of pulmonary artery occlusion pressure in pulmonary hypertension due to partial occlusion. AB - OBJECTIVE: To evaluate partial occlusion in patients with pulmonary hypertension with regard to a) the degree to which it leads to overestimation of pulmonary artery occlusion pressure (Ppao) and b) identification of factors that could enhance its recognition. DESIGN: Observational descriptive study. SETTING: Medical intensive care unit. PATIENTS: Fourteen patients with pulmonary hypertension and an increased pulmonary artery diastolic pressure (Ppad) - Ppao gradient (> or = 10 mm Hg). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The Ppao was recorded during partial occlusion (partial Ppao) and after catheter repositioning to obtain a lower, more accurate value (best Ppao). The error due to partial occlusion, defined as the difference between the partial Ppao and the best Ppao, was 13 +/- 5 mm Hg (range, 6-21 mm Hg). The previously widened Ppad - Ppao gradient invariably narrowed during partial occlusion and then increased by 13 +/- 5 mm Hg (range, 5-23) during the best Ppao measurement. There was a moderate correlation between the error due to partial occlusion (partial Ppao - best Ppao) and both the mean pulmonary artery pressure (r =.77, <.01) and the Ppad - Ppao gradient (r =.79, <.01). CONCLUSIONS: Partial occlusion in patients with pulmonary hypertension may lead to significant overestimation of the Ppao and should be suspected when there is a substantial increase in the Ppao without a concomitant increase in the Ppad, as reflected by a marked narrowing of a previously widened Ppad - Ppao gradient. PMID- 12545001 TI - Mortality prediction at admission to intensive care: a comparison of microalbuminuria with acute physiology scores after 24 hours. AB - OBJECTIVE: To compare low level albumin excretion (microalbuminuria), a marker of systemic capillary permeability, with mortality, Acute Physiologic And Chronic Health Evaluation (APACHE II) score, the Simplified Acute Physiologic (SAP II) score, and their derived mortality probabilities in patients admitted to a general intensive care unit. DESIGN: Prospective observational study. SETTING: A 14-bed intensive care unit in a university teaching hospital. PATIENTS: A total of 140 consecutive patients (59 surgical, 48 medical, 22 trauma, and 11 burns). INTERVENTIONS: Urine collection within 15 mins of intensive care unit admission for assessment of microalbuminuria. MEASUREMENTS AND MAIN RESULTS: Microalbuminuria, expressed as the albumin-creatinine ratio (ACR: normal, <2.3 mg/mmol), was compared with mortality, APACHE II and SAP II scores and their derived mortality probabilities after 24 hrs, intensive care unit stay, and markers of organ function and inflammation. Median (95% confidence interval) ACR at admission for survivors (n = 115) and nonsurvivors (n = 25) were 4.2 (3.6-6.5) and 17.8 (8.0-40.8) mg/mmol, respectively (p =.0002 Mann Whitney). For 92 surgical, trauma, and burn patients, of whom 81 survived, ACR of >5.9 mg/mmol gave a sensitivity for death of 100%, specificity of 59%, positive predictive value of 25%, and negative predictive value of 100%. Mortality probability receiver operator characteristic curve areas for ACR, APACHE II, and SAP II were 0.843 (p <.0001), 0.793 (p =.0004), and 0.770 (p =.0017), respectively. ACR was associated with intensive care unit stay (p =.0021) and highest serum C-reactive protein (p =.0002), serum creatinine (p <.0001), and bilirubin (p =.0009). For 48 medical patients, of whom 34 survived, admission ACRs for survivors and nonsurvivors were 8.3 (5.7-10.8) and 10.7 (4.1-48.2) mg/mmol, respectively (p =.32). SAP II, but not APACHE II, score was significantly higher for nonsurvivors. CONCLUSIONS: For surgical, trauma, and burn patients, but not medical patients, microalbuminuria within 15 mins of intensive care unit admission predicted death as well as APACHE II and SAP II scores calculated after 24 hrs, and it shows promise as a predictor of outcome. PMID- 12545002 TI - Outcome and early prognostic indicators in patients with a hematologic malignancy admitted to the intensive care unit for a life-threatening complication. AB - OBJECTIVES: To assess the outcome and to identify early prognostic indicators in a global population of patients with hematologic malignancy admitted to the intensive care unit for a life-threatening complication. DESIGN: Retrospective observational study. SETTING: Medical intensive care unit at a tertiary university hospital. PATIENTS: A total of 124 consecutive critically ill patients with a hematologic malignancy admitted to the intensive care unit during a 3.5-yr period. MEASUREMENTS We collected variables at admission and during admission and identified predictors of in-hospital mortality by stepwise logistic regression analysis. MAIN RESULTS: Mean Acute Physiology and Chronic Health Evaluation II score was 26 +/- 7.7. Sixty-one percent had a high-grade malignancy, and 27% had active disease. Thirty-five percent were leukopenic (leukocyte count, <1.0 x 10(9)/L) at admission. Respiratory failure (48%), sepsis (18.5%), and neurologic impairment (17%) were the major reasons for admission at the intensive care unit. Seventy-one percent of the patients required ventilatory support for a median duration of 6 (3-17) days, 46% received vasopressors at admission, and 26.6% needed renal replacement therapy during their intensive care unit stay. A recent bacteremia precipitating intensive care unit admission was found in 21.8% of the patients. Crude intensive care unit, in-hospital, and 6-month mortality rates were 42%, 54%, and 66%, respectively. Four variables were independently associated with outcome in a multivariate logistic regression analysis: leukopenia (odds ratio, 2.9; 95% confidence interval, 1.1-7.7), vasopressors (odds ratio, 3.74; 95% confidence interval, 1.4-9.8), and urea of >0.75 g/L (>12 mmol/L) (odds ratio, 9.4; 95% confidence interval, 4.2-26) at admission were associated with poor outcome, whereas recent bacteremia (odds ratio, 0.17; 95% confidence interval, 0.05-0.58) was associated with better prognosis. Using these variables, we arbitrarily categorized our population into three groups for survival analysis: a low-risk group (low urea with or without either leukopenia or vasopressors, n = 60), an intermediate-risk group (high urea or a combination of leukopenia and vasopressors, n = 34), and a high-risk group (high urea in combination with leukopenia or vasopressors, n = 27). Patients with a bacteremia prompting intensive care unit admission were allocated to a one-step-lower risk group. Survival probabilities at 30 days and 6 months were 75% and 55% in the first group, 35% and 21% in the second group, and 4% and 0%, respectively, in the third group ( <.001). CONCLUSION: The general reluctance to admit patients with a hematologic malignancy to the intensive care unit, even with severe critical illness, is unjustified. However, we identified four early predictors of outcome that may be of value in deciding in which patients advanced or prolonged support should not be continued. PMID- 12545003 TI - Noise, stress, and annoyance in a pediatric intensive care unit. AB - OBJECTIVE: To measure and describe hospital noise and determine whether noise can be correlated with nursing stress measured by questionnaire, salivary amylase, and heart rate. DESIGN: Cohort observational study. SETTING: Tertiary care center pediatric intensive care unit. SUBJECTS: Registered nurses working in the unit. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Eleven nurse volunteers were recruited. An audiogram, questionnaire data, salivary amylase, and heart rate were collected in a quiet room. Each nurse was observed for a 3-hr period during patient care. Heart rate and sound level were recorded continuously; saliva samples and stress/annoyance ratings were collected every 30 mins. Variables assessed as potential confounders were years of nursing experience, caffeine intake, patients' Pediatric Risk of Mortality Score, shift assignment, and room assignment. Data were analyzed by random effects multiple linear regression using Stata 6.0. The average daytime sound level was 61 dB(A), nighttime 59 dB(A). Higher average sound levels significantly predicted higher heart rates (p =.014). Other significant predictors of tachycardia were higher caffeine intake, less nursing experience, and daytime shift. Ninety percent of the variability in heart rate was explained by the regression equation. Amylase measurements showed a large variability and were not significantly affected by noise levels. Higher average sound levels were also predictive of greater subjective stress (p =.021) and annoyance (p =.016). CONCLUSIONS: In this small study, noise was shown to correlate with several measures of stress including tachycardia and annoyance ratings. Further studies of interventions to reduce noise are essential. PMID- 12545004 TI - Quality benefits of an intensive care clinical information system. AB - OBJECTIVE: This study was performed to quantify the quality benefits and staff perceptions of a computerized clinical information system implementation in an intensive care unit. Although clinical information systems have been available and implemented in many intensive care units for more than a decade, there is little objective evidence of their impact on the quality of care and staff perceptions. DESIGN: A longitudinal observational study before and after clinical information system implementation. SETTING: A 12-bed adult general intensive care unit in a large Australian tertiary referral teaching hospital. INTERVENTION: Implementation of a fully featured clinical information system to replace paper based charts of patient observations, clinical records, results reporting, and drug prescribing. MEASUREMENTS AND MAIN RESULTS: The frequency of clinical adverse events over a 4-yr period using an established reporting system was examined. Pre- and postimplementation staff questionnaires were distributed and analyzed. There were significant reductions in the rates of medication, intravenous therapy, and ventilator incidents. There was a trend toward a reduction in pressure sores. The survey, utilizing a validated questionnaire, demonstrated a positive perception of the clinical information system by nursing staff, with less time spent in documentation and more time in patient care. Nursing staff recruitment and retention improved after clinical information system implementation. CONCLUSIONS: Implementation of a fully featured clinical information system was associated with significant improvements in key quality indicators, positive nursing staff perceptions, and some positive resource implications. PMID- 12545005 TI - Critical care transesophageal endosonography and guided fine-needle aspiration for diagnosis and management of posterior mediastinitis. AB - BACKGROUND: Acute mediastinitis is a serious complication; it occurs after esophageal perforation and thoracic surgery and is rarely due to infections. Clinical and computed tomographic scan signs may be nonspecific, especially in postoperative patients. DESIGN: We prospectively evaluated the value of transesophageal endosonography with guided fine-needle aspiration in the diagnosis and identification of etiologic agents in critically ill patients with suspected posterior mediastinitis. SETTING: University hospital. PATIENTS AND METHODS: Transesophageal endosonography/fine-needle aspiration was performed at the bedside in the intensive care unit with a Pentax 34UX echo-endoscope and a portable Hitachi console (EUB 525). Eighteen patients with clinically suspected mediastinitis were examined with intensive care team support. RESULTS: Computed tomography was performed before transesophageal endosonography in all 18 patients and was inconclusive in 9. Transesophageal endosonography detected mediastinal lesions in 16 (89%) of 18 patients and was more accurately diagnostic than computed tomography (p =.0082). Fifteen patients had undergone surgery (11 esophagectomy, 1 other esophageal surgery, 1 head/neck cancer surgery, 1 complication after dilatational tracheostomy, and 1 with intervention after polytrauma). Three patients were suspected to have nonpostoperative mediastinitis. In 16 patients, infectious organisms were detected (bacterial, n = 14; fungal, n = 1; tuberculosis, n = 1). Culture and sensitivity of transesophageal endosonography/fine-needle aspiration specimens led to appropriate drug therapy. In two patients, methicillin-resistant Staphylococcus aureus was detected, leading to isolation care. Twelve patients improved; six died. Of the two patients in whom transesophageal endosonography did not detect a mediastinitis, one was false negative on autopsy. There were no complications. CONCLUSION: Bedside transesophageal endosonography/fine-needle aspiration of posterior mediastinal lesions in critically ill patients was an effective and relatively noninvasive way to detect mediastinitis and provide material to identify the etiologic agent. It was particularly useful in postesophagectomy patients. PMID- 12545006 TI - Association of tumor necrosis factor gene polymorphisms and prolonged mechanical ventilation after coronary artery bypass surgery. AB - OBJECTIVE: Prolonged mechanical ventilation is a common complication after coronary artery bypass graft surgery. Tumor necrosis factor alpha is an important proinflammatory mediator in the post-coronary artery bypass graft inflammatory cascade. We attempted to study the effect of polymorphisms at the -308 site in the promoter region of the tumor necrosis factor gene (TNF-308) and the +250 site within the lymphotoxin-alpha gene (LT alpha+250) on the risk of prolonged mechanical ventilation after coronary artery bypass grafting. DESIGN: Prospective observational study. SETTING: Tertiary care center. PATIENTS: A total of 400 patients undergoing coronary artery bypass grafting were enrolled. MEASUREMENTS: The primary end point was time to extubate. Secondary end points were the percentages of patients extubated at 8, 24, and 48 hrs; the length of intensive care unit and hospital stay; the need for a rehabilitation facility; and 30-day mortality. Precollected blood was used for gene analysis. Genotyping was performed by polymerase chain reaction and restriction enzyme digestion. MAIN RESULTS: Patients with an AA genotype at the LT alpha+250 site and those without the LT alpha+250G/-308TNFG haplotype had a shorter duration of mechanical ventilation (11.5 vs. 27.8 hrs and 11.2 vs. 29.4 hrs; =.039 and.01, respectively). The risk of prolonged mechanical ventilation at 8, 24, and 48 hrs was higher for patients with a GA or GG genotype at the LT alpha+250 site and the LT alpha+250G/TNF-308G haplotype. This association between genotype and duration of mechanical ventilation was more dramatic in patients undergoing conventional coronary artery bypass grafting than in those undergoing off-pump coronary artery bypass grafting. With Bayesian analysis, clinical criteria and genotype can be used sequentially to predict the risk of prolonged mechanical ventilation. CONCLUSIONS: The LT alpha+250 and LT alpha+250G/TNF-308G haplotypes are associated with prolonged mechanical ventilation after coronary artery bypass graft. Preoperative genetic screening may guide intraoperative management to reduce postoperative complications. PMID- 12545007 TI - Adrenal insufficiency during septic shock. AB - OBJECTIVE: To determine whether a baseline (random) cortisol concentration <25 microg/dL in patients with septic shock was a better discriminator of adrenal insufficiency than the standard (250 microg) and the low-dose (1 microg) corticotropin stimulation tests as assessed by the hemodynamic response to steroid replacement. SETTING: Intensive care unit. PATIENTS: Fifty-nine patients with septic shock. Their mean age was 57 +/- 16.7 yrs; 29 were male. INTERVENTIONS: A baseline cortisol concentration was obtained. Patients then received an intravenous injection of 1 microg of corticotropin (low-dose test) followed 60 mins later by an injection of 249 microg of corticotropin (high-dose test). Cortisol concentrations were obtained 30 and 60 mins after low- and high dose corticotropin. All patients were administered hydrocortisone (100 mg every 8 hrs) for the first 24 hrs while awaiting results of cortisol assessment. Patients were considered steroid responsive if the pressor agent could be discontinued within 24 hrs of the first dose of hydrocortisone. MEASUREMENTS AND MAIN RESULTS: Forty-seven percent of patients died. Twenty-two percent of patients met the diagnostic criteria of adrenal insufficiency by the low-dose test and 8% by the high-dose test. However, 61% of patients met the criteria of adrenal insufficiency when we used a baseline cortisol concentration of <25 microg/dL. Twenty-two patients (37%) were steroid responsive; the baseline serum cortisol was 14.1 +/- 5.2 microg/dL in the steroid-responsive patients compared with 33.3 +/- 18 microg/dL in the steroid-nonresponsive patients (p <.0001). Ninety-five percent of steroid-responsive patients had a baseline cortisol concentration <25 microg/dL. Fifty-four percent of steroid responders had a diagnostic low-dose test and 22% a diagnostic high-dose test. Receiver operating characteristic curve analysis revealed that a stress cortisol concentration of 23.7 microg/dL was the most accurate diagnostic threshold for determination of the hemodynamic response to glucocorticoid therapy. CONCLUSIONS: Adrenal insufficiency is common in patients with septic shock, the incidence depending largely on the diagnostic test and criteria used to make the diagnosis. There is clearly no absolute serum cortisol concentration that distinguishes an adequate from an insufficient adrenal response. However, we believe that a random cortisol concentration of <25 microg/dL in a highly stressed patient is a useful diagnostic threshold for the diagnosis of adrenal insufficiency. PMID- 12545008 TI - Comparison of black and white families' experiences and perceptions regarding organ donation requests. AB - CONTEXT: Black families donate at a lower rate than white families. To help develop effective interventions, we compared black and white families' experiences. OBJECTIVE: To compare the organ donation request experiences of black and white patients' families with the hope of identifying factors to better inform consent-rate interventions among blacks. DESIGN: Chart reviews were conducted on files of all deceased patients. Audiotaped in-person interviews were conducted with family members, and telephone interviews were conducted with involved healthcare providers and organ procurement organization staff. SETTING: Nine trauma hospitals located in southwest Pennsylvania and northeast Ohio. PARTICIPANTS: A sample of 415 families of organ donor-eligible patients (61 black, 354 white). MEASURES: Measures of families' attitudes about and experiences with donation and transplantation. RESULTS: White families were more likely to be correctly perceived as receptive to donation. Black families viewed as receptive were less likely to be engaged in discussing as many donation related issues as white families (p <.01). They were also less likely to have spoken to an organ procurement organization representative (p =.024) and were given fewer opportunities to consider the decision with healthcare provider or organ procurement organization staff. Black families had less knowledge about their family member's wishes, expressed less-favorable attitudes toward organ donation and the health care system, and, finally, were less likely than white families to donate organs (p =.001). A minority of black families (32.8%) knew about the need for more black donors. CONCLUSIONS: Interventions to increase consent among blacks need to encourage openness about organ donation within the black community and change healthcare provider and organ procurement organization attitudes and practices toward black families as potential donor families. PMID- 12545009 TI - Plasma concentration of Gc-globulin is associated with organ dysfunction and sepsis after injury. AB - OBJECTIVE: Clinical and experimental studies suggest that the proteins of the extracellular actin scavenger system have a role in the pathophysiological processes taking place in critically ill and injured patients. Circulating levels of Gc-globulin and gelsolin are reduced shortly after severe trauma, and admission levels of Gc-globulin are associated with survival. Herein, we sought to measure the association between admission levels of Gc-globulin and postinjury organ dysfunction and infection. We also wanted to describe the serial changes in Gc-globulin in these severely injured patients. DESIGN: Prospective cohort. SETTING: Intensive care unit at a county hospital that serves as a level one trauma center. PATIENTS: Ninety-eight consecutive trauma victims admitted to the intensive care unit for >24 hrs during a 4-month period. MEASUREMENTS AND MAIN RESULTS: Circulating levels of Gc-globulin were measured by using immunonephelometry. All patients were evaluated daily to obtain the necessary data for assessment of organ dysfunction and sepsis. The median Gc-globulin concentration at admission was 127 mg/L in patients who developed severe multiple organ dysfunction compared with 184 mg/L in patients who did not (p =.001). The admission level of Gc-globulin was comparable to known risk factors such as age and injury severity score, regarding development of organ dysfunction. Plasma concentrations of Gc-globulin remained significantly lower in patients who developed respiratory failure and sepsis, compared with patients who did not develop these complications (p =.02 and p=.015, respectively). CONCLUSIONS: Admission plasma concentration of Gc-globulin is lower in patients who develop organ dysfunction and sepsis after traumatic injury. These data, combined with the work of others, support the hypothesis that actin release and depletion of the extracellular actin scavenger system proteins are associated with, and may contribute in part to, the complications of sepsis and organ dysfunction, particularly respiratory failure and thrombocytopenia. PMID- 12545010 TI - Exacerbation with granulocyte colony-stimulating factor of prior acute lung injury during neutropenia recovery in rats. AB - OBJECTIVE: Neutropenia recovery may be associated with an increased risk of respiratory function deterioration. A history of pneumonia complicating neutropenia has been identified as the leading cause of adult respiratory distress syndrome during neutropenia recovery in patients receiving anticancer chemotherapy, suggesting that neutropenia recovery may worsen prior lung injury. DESIGN: Controlled animal study. SETTING: Research laboratory of an academic institution. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: We studied the effect of recovery from cyclophosphamide-induced neutropenia on endotoxin (lipopolysaccharide)- or hydrochloric acid-induced acute lung injury in rats. We also studied the effects of adding granulocyte colony-stimulating factor. MEASUREMENTS AND MAIN RESULTS: Compared with noncyclophosphamide-treated rats, rats undergoing neutropenia recovery had a higher wet/dry lung weight ratio after hydrochloric acid-induced but not lipopolysaccharide-induced acute lung injury. Granulocyte colony-stimulating factor significantly increased both alveolar cell recruitment (bronchoalveolar lavage fluid counts) and pulmonary edema (wet/dry lung ratio) in both acute lung injury models during neutropenia recovery. Furthermore, in an experiment in hydrochloric acid-instilled rats, exacerbation by granulocyte colony-stimulating factor of hydrochloric acid-induced acute lung injury was inhibited by lidocaine, which prevents adhesion of neutrophils to endothelial cells. Tumor necrosis factor-alpha and interleukin-1 beta concentrations in supernatants of lipopolysaccharide-stimulated alveolar macrophages from rats undergoing neutropenia recovery with granulocyte colony stimulating factor treatment were significantly increased compared with rats undergoing neutropenia recovery without granulocyte colony-stimulating factor. CONCLUSION: Neutropenia recovery can worsen acute lung injury, and this effect is exacerbated by granulocyte colony-stimulating factor. PMID- 12545011 TI - Hydrocortisone and the mitogen-activated protein kinase inhibitor U0126 acutely suppress reactive oxygen species generation from circulating granulocytes after gunshot injuries in the pig. AB - OBJECTIVE: Several external stimuli, including trauma, increase the endogenous production of reactive oxygen species that spontaneously attack vital biological molecules. In addition to their direct toxic effects, several secondary messenger systems are induced. To forestall a subsequent organ dysfunction, a short-term posttraumatic down-regulation of granulocyte function has been advocated. Corticosteroids are potent and universal anti-inflammatory agents, but they have well-known side effects. Modulation of the mitogen-activated protein kinase cascade is an alternative approach. The purpose of this study was to investigate how the posttraumatic production of reactive oxygen species can be modulated by hydrocortisone or the extracellular signal-regulated kinase inhibitor U0126. DESIGN: Prospective randomized trial. SETTING: Field hospital and research laboratory. SUBJECTS: Seventeen male pigs. INTERVENTIONS: In general anesthesia, the pigs were exposed to a standardized insult: one gunshot hitting the right femur from a distance of 25 m, and one pistol shot to the left upper abdomen from close range. Following immediate first aid treatment, the animals were transported to a nearby field hospital. According to randomization, the animals received either hydrocortisone 250 mg intravenously (group 1, n = 9) or a similar amount of saline (group 2, n = 8). The injections were given 5 mins after the last shot. Blood samples were drawn before shooting, immediately before hydrocortisone was given, and 60 mins after shooting. Circulating neutrophils were isolated, and the production of reactive oxygen species was measured fluorometrically. Neutrophils from nine randomly chosen animals (five from group 1 and four from group 2) were treated in vitro with the extracellular signal regulated kinase inhibitor U0126. MEASUREMENTS AND MAIN RESULTS: The injuries as evaluated by the abbreviated injury scale did not differ between the animals. All survived the first 60 mins. While the in vivo production of reactive oxygen species tended to increase in the controls, a significant reduction was measured in the hydrocortisone group. Subsequent treatment with U0126 further reduced the synthesis of reactive oxygen species by about two thirds in both groups, independently of time. CONCLUSIONS: Early injection of hydrocortisone after trauma inhibits the synthesis of reactive oxygen species from circulating neutrophils. Inhibition of the extracellular signal-regulated kinase branch of the mitogen-activated protein kinase signaling cascade is an alternative approach. The powerful in vitro capacity of selective extracellular signal regulated kinase inhibitors to reduce the posttraumatic reactive oxygen species generation deserves further investigations, and compelling evidence of their in vivo usefulness is still lacking. PMID- 12545012 TI - Application of a rat model of streptococcal shock to evaluate on-line hemoperfusion and removal of circulating superantigens. AB - OBJECTIVE: To evaluate the efficacy of on-line hemoperfusion for the removal of circulating superantigens in a rat model of streptococcal shock. DESIGN: In vitro and experimental animal studies. SETTING: University research laboratories. INTERVENTIONS: Chemically modified polystyrene-based composite fiber reinforced with polypropylene was formulated in discs and used to evaluate the removal of superantigenic toxins from culture supernatants in vitro, and from blood in vivo. MEASUREMENTS AND MAIN RESULTS: Incubation of streptococcal supernatant with a single disc reduced the concentration of the superantigen streptococcal pyrogenic exotoxin A from 90.9 +/- 12.7 ng/mL with the control fiber to 32.5 +/- 3.6 ng/mL with active fiber (p <.001). The active discs also brought about a dose-dependent reduction in mitogenic activity that was highly significant (counts reduced from 82,133 +/- 2747 using three control discs to 26,307 +/- 3547 with three active discs [p <.001]). Beginning 6 hrs after infection, animals were hemoperfused for 3 hrs over columns containing control or active fiber. At the end of the treatment period, there was a significant decrease in the number of circulating bacteria in the active group (3.5 x 10(4) vs. 3.1 x 10(3) colony-forming units/mL, p<.05). However, bacterial counts subsequently increased and by 15 hrs and at all subsequent time points, the number of circulating bacteria was no different between the two groups. There was a highly significant and sustained difference in circulating streptococcal pyrogenic exotoxin A levels between the groups. Streptococcal pyrogenic exotoxin A levels at 9 hrs were 19.9 ng/mL in the controls vs. 2.1 ng/mL in the active group (p =.05). Animals perfused over active fibers had a highly significant survival advantage compared with control or nonperfused groups (p <.01). CONCLUSIONS: Hemoperfusion and on-line removal of superantigens merits further study as a possible treatment strategy for streptococcal shock syndromes. The mechanism by which the fibers are operating requires further investigation. PMID- 12545013 TI - Combination of intravenously infused methylene blue and inhaled nitric oxide ameliorates endotoxin-induced lung injury in awake sheep. AB - OBJECTIVE: To evaluate the effects of a combination of methylene blue, an inhibitor of the nitric oxide pathway, and inhaled nitric oxide on endotoxin induced acute lung injury in awake sheep. DESIGN: Prospective, randomized, controlled experimental study. SETTING: University animal laboratory. SUBJECTS: Twenty-four yearling, awake sheep. INTERVENTIONS: The sheep were anesthetized and instrumented with vascular catheters. After 1 wk of recovery, the animals underwent tracheotomy and were subjected to intravenous infusions of endotoxin 10 ng x kg-1 x min-1 and isotonic saline 3 mL x kg-1 x hr-1 for 8 hrs. The sheep were randomly assigned to three groups of eight animals each: a) the control group received endotoxin and saline; b) the INO group received endotoxin, saline, and inhaled nitric oxide 40 ppm for 5 hrs; and c) the MB/INO group received endotoxin, saline, and methylene blue 3 mg/kg as an intravenous bolus injection followed by a continuous infusion of 3 mg x kg-1 x min-1 for 6 hrs in combination with inhaled nitric oxide 40 ppm for 5 hrs. MEASUREMENTS AND MAIN RESULTS: Hemodynamic variables and blood gases were determined hourly. In the early phase of endotoxemia (0-2 hrs), methylene blue/inhaled nitric oxide reduced the increments in pulmonary arterial pressure, pulmonary microvascular pressure, and pulmonary vascular resistance index by 60% compared with the controls and to a greater extent than did inhaled nitric oxide alone. During the late phase, all the preceding variables returned closely to baseline following inhaled nitric oxide or methylene blue/inhaled nitric oxide but remained remarkably elevated in the control group. Inhaled nitric oxide and methylene blue/inhaled nitric oxide reduced the increase in extravascular lung water by 40% and 80%, respectively. Inhaled nitric oxide transiently attenuated the increase in venous admixture and did not prevent a decrease in arterial oxygenation. In the methylene blue/inhaled nitric oxide group, blood gases remained unchanged from baseline. CONCLUSIONS: In sheep, methylene blue/inhaled nitric oxide protects more efficiently against acute lung injury than inhaled nitric oxide alone, as indicated by a milder pulmonary hypertension, less extravascular lung water accumulation, and maintained gas exchange. PMID- 12545014 TI - Albumin resuscitation increases cardiomyocyte contractility and decreases nitric oxide synthase II expression in rat endotoxemia. AB - OBJECTIVE: Hypotension and hypoperfusion during septic shock may contribute to tissue hypoxia and the intramyocardial inflammatory response that results in myocardial dysfunction. Therefore, we hypothesized that crystalloid or colloid resuscitation may alter myocardial dysfunction. DESIGN: Randomized, controlled, prospective animal study. SETTING: University animal laboratory. SUBJECTS: Sprague-Dawley rats (250-300 g, n = 6/group). INTERVENTIONS: Rats received an intraperitoneal injection of 10 mg/kg lipopolysaccharide or control. One hour later, rats were randomized to intravenous resuscitation and received either 30 mL/kg normal saline, 10 mL/kg 10% pentastarch, 10 mL/kg 5% rat albumin, or no volume. MEASUREMENTS AND MAIN RESULTS: We measured fractional shortening of cardiomyocytes isolated 5 hrs after lipopolysaccharide or control injection. In separate identical experiments, we measured myocardial interleukin-6, macrophage inhibitory protein-2, and nitric oxide synthase II protein and messenger RNA expression. Control fractional shortening of 24.1 +/- 2.2% was decreased by lipopolysaccharide to 18.8 +/- 1.2% (p <.001). Volume resuscitation after lipopolysaccharide significantly improved fractional shortening (p <.001). In particular, albumin resuscitation increased fractional shortening to 23.5 +/- 0.9%, which was more than either saline (fractional shortening 20.1 +/- 1.7%,p <.01) or pentastarch (fractional shortening 21.4 +/- 0.9%,p <.01). Myocardial macrophage inhibitory protein-2 protein and interleukin-6 and macrophage inhibitory protein-2 messenger RNA expression and neutrophil content were elevated following lipopolysaccharide (p <.05) but were not altered by volume resuscitation. Myocardial nitric oxide synthase II protein and messenger RNA expression increased following lipopolysaccharide (p <.01) and decreased with albumin resuscitation. CONCLUSIONS: We conclude that following lipopolysaccharide injection, volume resuscitation improves cardiomyocyte fractional shortening. Albumin resuscitation is particularly beneficial in preventing reduced cardiomyocyte contractility, and this benefit may be related to an albumin induced reduction in nitric oxide synthase II protein and messenger RNA expression following endotoxin injection. PMID- 12545015 TI - Mild hypothermia during hemorrhagic shock in rats improves survival without significant effects on inflammatory responses. AB - OBJECTIVE: To explore the hypothesis that the survival benefit of mild, therapeutic hypothermia during hemorrhagic shock is associated with inhibition of lipid peroxidation and the acute inflammatory response. DESIGN: Prospective and randomized. SETTING: Animal research facility. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: Rats underwent pressure-controlled (mean arterial pressure 40 mm Hg) hemorrhagic shock for 90 mins. They were randomized to normothermia (38.0 +/- 0.5 degrees C) or mild hypothermia (33-34 degrees C from hemorrhagic shock 20 mins to resuscitation time 12 hrs). Rats were killed at resuscitation time 3 or 24 hrs. MEASUREMENTS AND MAIN RESULTS: All seven rats in the hypothermia group and seven of 15 rats in the normothermia group survived to 24 hrs (p <.05). Hypothermic rats had lower serum potassium and higher blood glucose concentrations at 90 mins of hemorrhagic shock (p <.05). At resuscitation time 24 hrs, the hypothermia group had less liver injury (based on serum concentrations of ornithine carbamolytransferase and liver histology) and higher blood glucose than the normothermia group (p <.05). There were no differences in serum free 8 isoprostane (a marker of lipid peroxidation by free radicals) between the two groups at either baseline or resuscitation time 1 hr. Serum concentrations of interleukin- 1 beta, interleukin-6, and tumor necrosis factor-alpha peaked at resuscitation time 1 hr. Tumor necrosis factor-alpha concentrations were higher (p <.05) at resuscitation time 1 hr in the hypothermia group compared with the normothermic group. Serum cytokine concentrations were not different between survivors and nonsurvivors in the normothermia group. Serum cytokine concentrations returned to baseline values in both groups by 24 hrs. There were no differences in the number of neutrophils in the lungs or the small intestine between the groups. More neutrophils were found in the lungs at resuscitation time 3 hrs than at resuscitation time 24 hrs in both groups (p <.01). CONCLUSIONS: These data suggest that lipid peroxidation and systemic inflammatory responses to hemorrhagic shock are minimally influenced by mild hypothermia, although liver injury is mitigated and survival improved. Other mechanisms of benefit from mild hypothermia need to be explored. PMID- 12545016 TI - Endothelin receptor antagonist bosentan improves microcirculatory blood flow in splanchnic organs in septic shock. AB - OBJECTIVE: Splanchnic ischemia is believed to play an important role in the development of multiple organ dysfunction in septic shock. The vasoconstrictor peptide endothelin can produce an intense and sustained splanchnic vasoconstriction and is increased in sepsis. The aim of this investigation was to study the effects of an endothelin antagonist on microcirculatory blood flow in multiple abdominal organs during septic shock. DESIGN: Prospective, controlled animal study. SETTING: University-affiliated research laboratory. SUBJECTS: Fifteen anesthetized and mechanically ventilated pigs. INTERVENTIONS: Septic shock was induced by fecal peritonitis. After 120 mins of sepsis, eight animals received 10 mg/kg bosentan intravenously followed by an intravenous infusion at 5 mg x kg-1 x hr-1 whereas seven (controls) received isotonic saline. At 240 mins after induction of sepsis both groups received hydroxyethyl starch, 20 mL/kg intravenously, to convert hypodynamic septic shock to hyperdynamic sepsis. MEASUREMENTS AND MAIN RESULTS: Microcirculatory blood flow was measured simultaneously and continuously in the jejunal muscularis, pancreas, liver, kidney, skeletal muscle, and gastric, jejunal, and colon mucosa by using a multiple-channel laser Doppler flow meter. After 120 mins, all animals had developed signs of hypodynamic sepsis with decreased cardiac index, mean arterial blood pressure, and gastric mucosal pH. Microcirculatory blood flow in the pancreas and liver had decreased by 20% and in the jejunal muscularis by >40% (p <.01) whereas it remained virtually unchanged in the gastric, jejunal, and colonic mucosa. After 240 mins, cardiac index, mean arterial blood pressure, gastric mucosal pH, and microcirculatory blood flow in the gastric mucosa, colon mucosa, jejunal muscularis, and pancreas had all deteriorated in the controls, whereas in the bosentan-treated group, cardiac index and microcirculatory blood flow in the pancreas, gastric, and colon mucosa improved. During hyperdynamic sepsis, cardiac index increased above baseline in both groups but significantly more in the bosentan group. In the control group, microcirculatory flow returned to baseline in most tissues except in skeletal muscle and jejunal muscularis. In the bosentan group, microcirculatory flow returned to or increased above baseline in all tissues except in the muscularis of the jejunum. CONCLUSIONS: The endothelin receptor antagonist bosentan significantly improved microcirculatory blood flow in many splanchnic organs and in peripheral tissues during septic shock. The results of this study are consistent with the hypothesis that endothelin plays an important role in the regulation of microcirculatory blood flow in splanchnic as well as in peripheral tissues during septic shock. PMID- 12545017 TI - Linkage disequilibrium between tumor necrosis factor (TNF)-alpha-308 G/A promoter and TNF-beta NcoI polymorphisms: Association with TNF-alpha response of granulocytes to endotoxin stimulation. AB - OBJECTIVE: Controversial data have been reported on the association between the tumor necrosis factor (TNF)-alpha-308 G[U279C]A promoter polymorphism or the TNF alpha I polymorphism with TNF-alpha plasma concentrations. The purpose of this study was to evaluate whether there is a linkage disequilibrium between the two polymorphisms. Moreover, the influence of these polymorphisms on the TNF-alpha synthesis of activated granulocytes was studied. DESIGN: Analysis of TNF-alpha concentrations of human whole blood after endotoxin stimulation. SETTING: Medical research laboratory. PATIENTS: Healthy human volunteers. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Healthy human volunteers were genotyped for both TNF polymorphisms by means of polymerase chain reaction. TNF-alpha plasma concentrations were determined with chemiluminescence after incubation of whole blood with endotoxin. A strong (p <.0001) linkage disequilibrium was found for the TNF-beta I and the TNF-alpha-308 genetic polymorphisms. Almost all individuals homozygous for the TNF-B2 allele of the TNF-beta I polymorphism were also TNF-alpha-308 G homozygotes. Carriers of the TNF-alpha-308 genotype AG had a significantly higher TNF-alpha production capacity than G homozygotes. The TNF beta I genotype TNF-B1/TNF-B2 was associated with significantly higher TNF-alpha concentrations than the genotype TNF-B2/TNF-B2. Individuals homozygous for the TNF-B2 and the TNF-alpha-308 G alleles had a significantly reduced TNF-alpha response compared with individuals heterozygous for both TNF polymorphisms. CONCLUSIONS: A linkage disequilibrium between the two TNF polymorphisms was found. This study revealed a significant association between genotype and phenotype for both TNF polymorphisms. Heterozygosity for both TNF polymorphisms is associated with an increased TNF-alpha response. PMID- 12545018 TI - Low-dose terlipressin improves systemic and splanchnic hemodynamics in fluid challenged endotoxic rats. AB - OBJECTIVE: Vasopressin has been used to treat arterial hypotension associated with hyperdynamic vasoplegic states, but detrimental effects on splanchnic circulation have been reported. We tested the effects of a low-dose vasopressin analogue, terlipressin (6 microg/kg), on systemic and splanchnic hemodynamics in fluid-challenged endotoxic rats (lipopolysaccharide, 30 mg/kg in 1 hr). DESIGN: Prospective, randomized, controlled experimental study with repeated measures. SETTING: Investigational animal laboratory. SUBJECTS: A total of 77 rats were divided into five groups: group C, control (17 rats); group E, LPS (18 rats); group EF, LPS plus fluid challenge (18 rats); group EFT, LPS plus fluid challenge plus terlipressin (18 rats); and group ET, LPS plus terlipressin (seven rats). INTERVENTIONS: Rats were anesthetized, mechanically ventilated, and instrumented to measure heart rate, mean arterial pressure, and abdominal aortic and mesenteric vein indexed blood flows; ileal microcirculation was assessed by laser Doppler. After LPS infusion, rats experienced an endotoxic shock and were resuscitated after the allocation group. The fluid challenge was targeted to maintain mean arterial pressure of >90 mm Hg and aortic blood flow at baseline values. MEASUREMENTS AND MAIN RESULTS: Terlipressin significantly (p <.05) increased mean arterial pressure without decreasing indexed aortic blood flow and heart rate in the fluid-challenged endotoxic rats (EFT) compared with EF rats and had detrimental effects in hypodynamic endotoxic rats (ET). Fluid challenge significantly (p <.05) increased mesenteric vein blood flow in both the EF and EFT groups, and terlipressin had no detrimental effect on mesenteric blood flow. Terlipressin significantly (p <.05) increased ileal microcirculation in fluid challenged endotoxic rats (EF and EFT) but not in hypodynamic endotoxic rats (E and ET). CONCLUSION: Low-dose terlipressin in fluid-challenged endotoxic rats improved systemic and splanchnic hemodynamics and improved the ileal microcirculation. PMID- 12545019 TI - Protegrin-1 increases bacterial clearance in sepsis but decreases survival. PMID- 12545020 TI - Determinants of tidal volume during high-frequency oscillation. AB - OBJECTIVE: High-frequency oscillation has been proposed for use in adult acute respiratory distress syndrome. However, limited data are available on the effect of pressure amplitude and rate (Hz) on tidal volumes delivered during high frequency oscillation in adults. DESIGN: Prospective, animal model, lung injury study. SETTING: Large-animal laboratory of a university-affiliated medical center. SUBJECTS: Nine sheep (29.2 +/- 2.4 kg). INTERVENTIONS: Severe lung injury was induced by repeated saline lung lavage. After stabilization, high-frequency oscillation was initiated at a mean airway pressure equal to the point of maximum curvature on the deflation limb of the pressure-volume curve (26 +/- 1.9 cm H2O). Tidal volume at all combinations of rates of 4, 6, 8, and 10 Hz, pressure amplitudes of 30, 40, 50, and 60 cm H2O, and inspiratory/expiratory ratios of 1:1 and 1:2 (using the Sensormedics 3100B oscillator) were measured. Flow was measured by a pneumotachometer, amplified and digitized at 1000 Hz. Three breaths were analyzed at each setting. MEASUREMENTS AND MAIN RESULTS: At both inspiratory/expiratory ratios, tidal volume was directly proportional to pressure amplitude and inversely proportional to frequency. During an inspiratory/expiratory ratio of 1:1, at 60 cm H2O pressure amplitude and 4 Hz, a tidal volume of 129.1 +/- 34.8 mL (4.4 +/- 1.2 mL/kg) was delivered. CONCLUSIONS: At low rates and high-pressure amplitudes in this model, tidal volumes approaching conventional mechanical ventilation can be delivered during high frequency oscillation. PMID- 12545021 TI - Genetic background conditions the effect of sex steroids on the inflammatory response during endotoxic shock. AB - OBJECTIVE: The contribution of gender to the mortality and morbidity of trauma patients is controversial. In addition, a genetic contribution has been recently indicated. The influence of these two variables was studied in a murine model of endotoxemia. DESIGN: Prospective, controlled, and randomized animal study. SETTING: A university research laboratory. SUBJECTS: Female and male mice (6-8 wks old) were injected with lipopolysaccharide (15 mg/kg). Additionally, mice were gonadectomized and supplemented with 5-alpha-dihydrotestosterone (357 mg/day), 17-beta-estradiol (23.8 microg/day), or placebo for 21 days and injected with lipopolysaccharide. Tumor necrosis factor-alpha was measured in plasma samples obtained after 1.5 hrs of lipopolysaccharide injection. MEASUREMENTS AND MAIN RESULTS: Higher tumor necrosis factor-alpha plasma levels were observed in C57BL/6J (B6) female mice as compared with males. Because this phenotype is not sex linked, we evaluated the role of sex steroids. Castrated male B6 mice showed higher lipopolysaccharide-induced tumor necrosis factor-alpha plasma levels than nonoperated controls. These lipopolysaccharide-induced tumor necrosis factor alpha levels were further increased after the administration of 17-beta-estradiol to castrated B6 male mice as compared with nonoperated male or female mice. In addition, 17-beta-estradiol-supplemented castrated mice showed a higher frequency of mortality than castrated males without hormone replacement or nonoperated mice. Analysis of castrated male mice from other strains (A/J, DBA/2J, AKR/J, BALB/cJ) supplemented with 17-beta-estradiol presented the opposite effect, a reduction in lipopolysaccharide-induced tumor necrosis factor-alpha plasma levels. CONCLUSIONS: These results suggest that sex steroids can modulate the inflammatory response and the outcome after injury in mice. The effect of sex steroids depends on the genetic background. PMID- 12545023 TI - Temperature sensitivity of glycolysis during sepsis. AB - OBJECTIVE: To investigate the temperature sensitivity of glycolysis during sepsis. DESIGN: A prospective, randomized, controlled animal study. SETTING: The Physiological Department of a University Hospital. SUBJECTS: Ten male Sprague Dawley rats, weighing 400-500 g. INTERVENTIONS: The rats were assigned to either a septic (n = 5) or a sham-control group (n = 5). After anesthesia (H0), experimental sepsis was induced by a cecal ligation and perforation, and the left lateral gastrocnemius was sampled. Four hours later (H4), a second anesthesia was performed to sample the contralateral muscle. The sham-control group underwent the same procedures, but the cecum was neither ligated nor incised. MEASUREMENTS AND MAIN RESULTS: Glycolytic flux (J(B), the rate at which glycogen can be used in muscle) and the transition time (t99 : the time required for the transition from aerobic to anaerobic metabolism) were measured by using spectrophotometry. The measurements were performed at seven different temperature levels, ranging from 32 to 42 degrees C. For each measured variable, the temperature sensitivity of glycolysis was assessed by computing the Q10 values, which is the variation ratio of the measured variable, attributed to a 10 degrees C temperature increase. In control rats, anesthesia and surgical procedures induced a J(B) increase (7.9 +/- 1.6 at H0 vs. 11.9 +/- 2.1 micromol x min-1 x g(tissue) at H4, p<.05) without any t99 variation. Whatever the group (control or septic), the same temperature variation induced an effect that was approximately three times higher in the hypothermia (<37 degrees C) than in the hyperthermia range (>37 degrees C; p<.05). However, a loss in thermal sensitivity was observed in septic rats in the hyperthermia range (Q10 = 1.2 +/- 0.1 for septic animals vs. 2.3 +/- 0.4 for control animals; p<.05). CONCLUSIONS: This study demonstrates that glycolysis is more sensitive to temperature in the hypothermia range than in the hyperthermia range. The loss in thermal sensitivity at >37 degrees C in septic rats suggests that sepsis may induce a dysregulation of glycolysis. From an energetic point of view, this signifies that hyperthermia may by itself impair energy metabolism without improving energy production and thus must be treated during sepsis. PMID- 12545022 TI - Modulation of serum cytokine levels by a novel superoxide dismutase mimetic, M40401, in an Escherichia coli model of septic shock: correlation with preserved circulating catecholamines. AB - OBJECTIVES: We have shown previously that inactivation of catecholamines by superoxide anions contributes to the loss of vascular reactivity to norepinephrine and the subsequent hypotension that develops in Gram-negative endotoxic shock. In addition to their vasopressor actions, catecholamines, via beta-adrenoceptor activation, are important regulators of cytokine production. Here we examined if maintenance of serum catecholamine levels by the superoxide dismutase mimetic, M40401, modulates serum cytokine levels and arterial hypotension in an Escherichia coli-infected conscious rat model of septic shock. DESIGN: Controlled laboratory animal study. SETTING: University animal research laboratory. SUBJECTS: Pathogen-free male Sprague-Dawley rats (n = 51). INTERVENTIONS: Conscious, antibiotic-treated animals with chronic in-dwelling carotid arterial and jugular venous catheters were intravenously infected with 10(10) live E. coli bacteria (O55:B5, n = 51) over 30 mins, ending at time = 0 hrs. At 0.5 or 3 hrs, infected rats were administered an intravenous infusion of either M40401 (n = 33) or 0.9% saline (n = 18) for 6 hrs at a rate of 1 mL/h. In additional experiments, anesthetized animals with catheterized left femoral arteries and veins were administered a dose-range of norepinephrine (0.1-1 microg/kg) as bolus intravenous injections. Thereafter, E. coli lipopolysaccharide (4 mg/kg, n = 6) was administered as a 0.3-mL slow bolus intravenous injection. One hour later, the norepinephrine protocol was repeated, after which the rats were administered an intravenous infusion of either M40401 or 0.9% saline for 15 mins. At 2 hrs, the dose response to norepinephrine was repeated. MEASUREMENTS AND MAIN RESULTS: Rats infected with live E. coli exhibited a biphasic fall in mean arterial pressure, with mortality reaching 83% by 24 hrs. Rats treated with M40401 (0.25, 2.5, or 25 microg x kg-1 x hr-1 ) 3 hrs after bacteremic sepsis maintained a normal mean arterial pressure, and mortality was dose-dependently reduced to 44, 33, and 22%, respectively, at 24 hrs. Furthermore, serum catecholamine levels were diminished in E. coli-infected rats treated with saline compared with rats treated with M40401. In separate experiments, E. coli-infected rats were administered M40401 (25 microg x kg-1 x hr-1 ) 0.5 hr after bacterial challenge. Blood samples taken at 0, 1.5, 3.5, and 6 hrs were analyzed for tumor necrosis factor-alpha, interleukin (IL)-1 beta, IL 6, and IL-10 and for norepinephrine and epinephrine. Serum levels of tumor necrosis factor-alpha and IL-1 beta were significantly depressed in M40401 treated septic rats, whereas IL-10 was elevated. Moreover, serum catecholamine levels were greater in M40401-treated septic rats at the same time points. IL-6 levels were unaffected by M40401 treatment. Finally we examined whether treatment with M40401 could reverse the hyporeactivity to norepinephrine typifying early septic shock. Using the E. coli lipopolysaccharide (4 mg/kg) challenged anesthetized rat model of shock, we demonstrated that the vasoconstrictor ability of norepinephrine was indeed restored after M40401 treatment (25 microg/kg). CONCLUSION: Postinfection treatment with the superoxide dismutase mimetic M40401 protects against hypotension, vascular hyporeactivity to catecholamines, and mortality associated with septic shock. Such beneficial effects correlate with both reduced oxidative inactivation of serum catecholamines and a reduction in canonical cytokine mediators of inflammation. PMID- 12545024 TI - Effect of in vitro hemodilution with hydroxyethyl starch and dextran on the activity of plasma clotting factors. AB - OBJECTIVE: A recent thrombelastography study indicated a compromised in vitro blood coagulation after 10:10 (equal parts of blood and infusion) and 10:4 (10 parts blood to four parts infusion) hemodilution with several plasma substitutes. Oncovertin N (Oncovertin) (a 10% dextran 40 solution) had the strongest anticoagulant effect of all solutions tested, and HAES-sterile 10% (HAES) (a 10% hydroxyethyl starch 200/0.5 solution) showed the strongest effect of five different hydroxyethyl starch preparations. The aim of this study was to determine how in vitro hemodilution with HAES and Oncovertin affects the activity of coagulation factors. DESIGN: HAES and Oncovertin were tested to determine the intrinsic effect of colloid molecules, as opposed to hemodilution. Normal saline (NaCl) and nonlactated Ringer solution were used as noncolloidal controls. SETTING: University research institute. PATIENTS: Six healthy volunteers. INTERVENTIONS: Twenty milliliters of blood was obtained from each subject. MEASUREMENTS AND MAIN RESULTS: Prothrombin index, activated partial prothrombin time, soluble fibrin monomers, and the activity of coagulation factors I, II, V, VII, VIII, IX, X, XI, and XII were measured with the Behring Chromotimer according to the manufacturer's instructions. Two dilution ratios of citrated blood to infusion were used: 10:10 (equal parts of blood and infusion) and 10:4 (10 parts blood to four parts infusion). Baseline was undiluted. Hemodilution with NaCl at both 10:4 and 10:10 influenced the coagulation variables measured. The activities of factors I, VII, and soluble fibrin monomers were less influenced than expected by hemodilution alone. The activities of factors II, V, IX, and XI were significantly (p <.04) lower with both 10:4 and 10:10 dilution with NaCl. In the assays for factors IX, XI, and XII, clots formed immediately after adding the appropriate reagents in the presence of Ringer solution at 10:10 hemodilution, so that the activities of those factors could not be measured. For the other factors and for 10:4 dilution, the outcome after Ringer solution was similar to that of NaCl. The activities were less influenced after 10:4 hemodilution with both HAES and Oncovertin than after dilution with NaCl and Ringer solution, with no significant differences from baseline. At 10:10 hemodilution with both HAES and Oncovertin, several factor activities were significantly (p <.04) lower than baseline. CONCLUSIONS: Both NaCl and Ringer solution cause measurable effects on coagulation factors at 10:4 hemodilution that can be explained by hemodilution alone. The effects on clotting factors of 10:4 hemodilution with HAES and Oncovertin were not significant. Even at 10:10 hemodilution with HAES or Oncovertin, the reduction in factor activities, although significantly (p <.04) different from baseline, was less than what was expected by dilution alone. PMID- 12545025 TI - Quantitative evaluation of the neuroprotective effects of hypothermia ranging from 34 degrees C to 31 degrees C on brain ischemia in gerbils and determination of the mechanism of neuroprotection. AB - OBJECTIVE: The present study was designed to determine whether the predominant factor responsible for neuroprotection of hypothermia ranging from 31 to 34 degrees C is prolongation of onset of ischemic depolarization or suppression of neuronal injury during ischemic depolarization and to quantitatively determine the neuroprotective effects of hypothermia of 34 degrees C and 31 degrees C. DESIGN: Prospective animal study. SETTING: A university research laboratory. SUBJECTS: Eighty-nine gerbils. INTERVENTIONS: Bilateral common carotid arteries were occluded for 3-20 mins. The brain temperature was set at 37 degrees C, 34 degrees C, or 31 degrees C before and during ischemic depolarization. MEASUREMENTS AND MAIN RESULTS: DC potentials were measured in the CA1 region, where histologic evaluation was performed 7 days later. Onset times of ischemic depolarization were 1.3 +/- 0.2, 1.6 +/- 0.4, and 2.4 +/- 0.7 mins at 37 degrees C, 34 degrees C, and 31 degrees C, respectively. The logistic regression curve demonstrated a close relationship between duration of ischemic depolarization and neuronal damage and showed a rightward shift by lowering the brain temperature. In the 37 degrees C, 34 degrees C, and 31 degrees C groups, the durations of ischemic depolarization causing 50% neuronal damage were estimated to be 8.0, 14.2, and 26.0 mins, respectively, and the ischemia times causing 50% neuronal damage were estimated to be 4.9, 8.1, and 14.2 mins, respectively. CONCLUSIONS: The onset of ischemic depolarization was prolonged in the 34 degrees C and 31 degrees C groups by only 0.3 and 1.1 mins, respectively, compared with that in the 37 degrees C group. Most of the neuroprotection by hypothermia was attributed to the suppression of neuronal injury during ischemic depolarization, suggesting that hypothermia has neuroprotective effects if it is initiated during the ischemic depolarization period. The results also indicate that the neuroprotective effect at 31 degrees C is about three times greater than that at 34 degrees C and that neuronal cells can withstand 2.9 times longer duration of ischemia at 31 degrees C than at 37 degrees C. PMID- 12545026 TI - Estimation of pulmonary artery occlusion pressure by an artificial neural network. AB - OBJECTIVE: We hypothesized that an artificial neural network, interconnected computer elements capable of adaptation and learning, could accurately estimate pulmonary artery occlusion pressure from the pulsatile pulmonary artery waveform. SETTING: University medical center. SUBJECTS: Nineteen closed-chest dogs. INTERVENTIONS: Pulmonary artery waveforms were digitally sampled before conventional measurements of pulmonary artery occlusion pressure under control conditions, during infusions of serotonin or histamine, or during volume loading. Individual beats were parsed or separated out. Pulmonary artery pressure, its first time derivative, and the beat duration were used as neural inputs. The neural network was trained by using 80% of all samples and tested on the remaining 20%. For comparison, the regression between pulmonary artery diastolic pressure and pulmonary artery occlusion pressure was developed and tested using the same data sets. As a final test of generalizability, the neural network was trained on data obtained from 18 dogs and tested on data from the remaining dog in a round-robin fashion. MEASUREMENTS AND MAIN RESULTS: The correlation coefficient between the pulmonary artery diastolic pressure estimate of pulmonary artery occlusion pressure and measured pulmonary artery occlusion pressure was.75, whereas that for the neural network estimate of pulmonary artery occlusion pressure was.97 (p <.01 for difference between pulmonary artery diastolic pressure and pulmonary artery occlusion pressure estimates). The pulmonary artery diastolic pressure estimate of pulmonary artery occlusion pressure showed a bias of 0.097 mm Hg (limits of agreement -7.57 to 7.767 mm Hg), whereas the neural network estimate of pulmonary artery occlusion pressure showed a bias of -0.002 mm Hg (-2.592 to 2.588 mm Hg). There was no significant change in the bias of the neural network estimate over the range of values tested. In contrast, the bias for the pulmonary artery diastolic pressure estimate significantly increased with the increasing magnitude of the pulmonary artery occlusion pressure. During round-robin testing, the neural network estimate of pulmonary artery occlusion pressure showed suboptimal performance (correlation coefficient between estimated and measured pulmonary artery occlusion pressure.59). CONCLUSIONS: A neural network can accurately estimate pulmonary artery occlusion pressure over a wide range of pulmonary artery occlusion pressure under conditions that alter pulmonary hemodynamics. We speculate that artificial neural networks could provide accurate, real-time estimates of pulmonary artery occlusion pressure in critically ill patients. PMID- 12545027 TI - Tissue oxygen reactivity and cerebral autoregulation after severe traumatic brain injury. AB - OBJECTIVE: To study the relationship between arterial blood pressure, intracranial pressure, directly measured brain tissue oxygenation (PtiO2), and middle cerebral artery blood flow velocity in severely head-injured patients. DESIGN: Prospective study. SETTING: Neurosurgical intensive care unit. PATIENTS A total of 14 patients with severe head injury. INTERVENTIONS: Pharmacologic blood pressure manipulations using norepinephrine. MEASUREMENTS AND MAIN RESULTS: We assessed the magnitude of PtiO2 related to changes in cerebral perfusion pressure in 12 of the patients. We calculated in all the static rate of regulation, which is an index to describe the change of cerebrovascular resistance, using cerebral artery blood flow velocity in relation to changing cerebral perfusion pressure. Finally, we calculated the rate of change in PtiO2, which quantifies the percentage of change in PtiO2 divided by the percentage of change in cerebral perfusion pressure. It is a new marker for cerebral tissue oxygen regulation based on direct measurement of PtiO2. There was a plateau phase for the cerebral perfusion pressure-PtiO2 relation that was similar to the autoregulatory plateau seen in the relationship between cerebral perfusion pressure and cerebral artery blood flow velocity. The rate of change in PtiO2 demonstrated a significant correlation with the static rate of regulation (R = -.61, <.05). A decrease in intracranial pressure when arterial blood pressure increased from 70 to 90 mm Hg was strongly correlated with static rate of regulation (R =.79, <.001). CONCLUSIONS Cerebral tissue PO2 demonstrates a plateau phase similar to what is known about cerebral blood flow velocity, which suggests a close link between cerebral blood flow and oxygenation. Static cerebral autoregulation is significantly correlated with cerebral tissue oxygen reactivity. PMID- 12545029 TI - Early predictors of mechanical ventilation in Guillain-Barre syndrome. AB - OBJECTIVE: Although mechanical ventilation is required in 30% of patients with Guillain-Barre syndrome, early predictors of this treatment remain unknown. DESIGN: Analysis of two cohorts of patients enrolled in two multicenter randomized clinical trials. SETTING: French intensive care and neurologic units. MEASUREMENTS: Demographic, neurologic, and biologic data; vital capacity; and time of onset, admission, and endotracheal mechanical ventilation were collected. RESULTS: Amid 722 consecutive adults not ventilated at admission, endotracheal mechanical ventilation was required in 313 (43%) patients. Multivariate analyses identified six predictors of endotracheal mechanical ventilation: time from onset to admission of <7 days (odds ratio, 2.51), inability to cough (odds ratio, 9.09), inability to stand (odds ratio, 2.53), inability to lift the elbows (odds ratio, 2.99) or head (odds ratio, 4.34), and liver enzyme increases (odds ratio, 2.09). In the 196 (27%) patients whose vital capacity was measured, time from onset to admission of <7 days (odds ratio, 5.00), inability to lift the head (odds ratio, 5.00), and vital capacity <60% (odds ratio, 2.86) predicted endotracheal mechanical ventilation. CONCLUSIONS: Patients with at least one of these predictors should be monitored in an intensive care unit. Mechanical ventilation was required in >85% of patients with at least four predictors from the first multivariate model and in 85% of patients with all three predictors from the second multivariate model. In these patients at high risk of respiratory failure, the value of early mechanical ventilation may deserve investigation. PMID- 12545028 TI - Timing of neurologic deterioration in massive middle cerebral artery infarction: a multicenter review. AB - OBJECTIVE: To determine the time interval between symptom onset and neurologic deterioration related to cerebral edema in patients with massive middle cerebral artery infarction. The time period between onset and neurologic deterioration represents the window for surgical intervention. DESIGN: Multicenter retrospective chart review. SETTINGS: Five university-affiliated medical centers. PATIENTS: Fifty-three patients with massive middle cerebral artery infarction who experienced neurologic deterioration defined by a decrease in the Glasgow Coma Scale score of two or more points attributable to mass effect. MEASUREMENTS AND MAIN RESULTS: A total of 53 patients (mean age, 62 +/- 18 yrs; 25 [47%] were men) with neurologic deterioration were identified by using International Classification of Diseases (9th revision) codes and local registries. Medical records and neuroimaging studies were reviewed by a stroke neurologist or neurointensivist to identify the time of neurologic deterioration. Thrombolytics were used at presentation in 19 (35%) patients. A total of 19 (36%) patients had neurologic deterioration within 24 hrs of symptom onset. By 48 hrs, 36 (68%) patients had manifested clinical deterioration. A few patients had later neurologic deterioration on day 3 (n = 10), day 4 (n = 2), day 5 (n = 2), and day 6 or after (n = 3). A total of 25 (47%) of the 53 patients died during hospitalization. The highest frequency of deaths occurred on day 3. CONCLUSIONS: Neurologic deteriorations related to cerebral edema after massive middle cerebral artery infarction occur in most patients within 48 hrs of symptom onset. PMID- 12545030 TI - Changes in biochemical and biophysical surfactant properties with cardiopulmonary bypass in children. AB - OBJECTIVE: The aim of the present study was to characterize pulmonary surfactant properties in children undergoing cardiovascular surgery with cardiopulmonary bypass. DESIGN: Prospective clinical trial. SETTING: University hospital pediatric intensive care unit. PATIENTS: Fifty pediatric patients with congenital cardiac defects undergoing cardiovascular surgery with (n = 35) and without (n = 15) cardiopulmonary bypass procedure. INTERVENTIONS: Tracheal aspirates were collected by saline lavage during routine suctioning before (baseline) and after cardiopulmonary bypass, as well as 4, 8, and 24 hrs after admission to the pediatric intensive care unit. MEASUREMENTS AND MAIN RESULTS: Total protein and phospholipid concentrations were assessed in native tracheal aspirates, in large surfactant aggregates, and in small surfactant aggregates. Phospholipid profiles and phosphatidylcholine fatty acids; surfactant apoproteins SP-A, SP-B, and SP-C (enzyme-linked immunosorbent assay); and surface activity (Pulsating Bubble Surfactometer) were all analyzed in large surfactant aggregates. With cardiopulmonary bypass, an initial increase in total protein content was followed by an increase in phospholipid concentration in tracheal aspirates. Large surfactant aggregates decreased 4 hrs after cardiopulmonary bypass (4 hrs, 22.6 +/- 5.6%; mean +/- SEM; p<.01 compared with baseline, 55.4 +/- 9.2%) but recovered within 24 hrs. The phospholipid-protein ratio of large surfactant aggregates 24 hrs after cardiopulmonary bypass (1.2 +/- 0.2; p<.01) was significantly decreased compared with baseline (2.9 +/- 0.6). The relative amount of phosphatidylglycerol content in the large surfactant aggregates-fraction dropped linearly over time but other phospholipids remained mainly unchanged. Phosphatidylcholine fatty acid profiles remained unaffected by cardiopulmonary bypass. The relative content of SP-B and SP-C in large surfactant aggregates increased approximately three-fold compared with baseline. Altogether, our findings with recovered large surfactant aggregate/small surfactant aggregate ratios and increased phospholipid in tracheal aspirates after 24 hrs represent an approximately ten-fold increase in large surfactant aggregate-associated SP-B and SP-C compared with baseline. Only minor changes were detected in biophysical properties of large surfactant aggregates throughout the observation period. CONCLUSIONS: Cardiopulmonary bypass procedure in children induces profound changes in the surfactant system involving both phospholipid and protein components; biophysical function may have been maintained by compensatory increase in SP-B and SP-C. PMID- 12545031 TI - Patient safety and the science of prevention: the time for implementing the Guidelines for the prevention of intravascular catheter-related infections is now. PMID- 12545032 TI - Survey of current status of intensive care teaching in English-speaking medical schools. AB - OBJECTIVE: To identify a consensus of opinion regarding the content of an intensive care core syllabus for undergraduate medical students and factors that may limit its teaching. DESIGN: Cross-sectional postal survey containing 35 items ranging from department structure to curriculum content and factors that limit the teaching of intensive care. SETTING: English-speaking medical schools (n = 210) listed in the 1986 World Health Organization Directory. MEASUREMENTS AND MAIN RESULTS: Of 122 (58%) returned questionnaires, a 45% return was achieved from the United States and 86% from non-U.S. countries. Most respondents (84%) considered teaching undergraduate intensive care to be essential; however, teaching intensive care was compulsory in only 31% of schools. Many schools (43%) reported recent changes to their intensive care curriculum. Most respondents (60%) thought that intensive care specialists should teach and that each student required a median (interquartile range) of 20 (10-80) hrs of teacher contact time. Resuscitation skills were taught in 98% of schools. In comparison, 63% of schools had no intensive care syllabus. More than 90% of respondents thought that the intensive care syllabus should include the following: cardiopulmonary resuscitation, assessment and management of the acutely ill patient; management of respiratory, circulatory, and multiple organ system failure (including systemic inflammatory response syndrome and sepsis); management of the unconscious patient; early postoperative care; and communication skills and ethics as they relate to end-of-life issues. Factors that limited intensive care teaching were lack of staff, funding, and time dedicated to teaching and excessive clinical workload. Student performance in intensive care was assessed by 66% of schools, but only 28% used a written or oral examination. CONCLUSIONS: By surveying a wide range of medical schools internationally, we have been able to define an undergraduate intensive care syllabus that could be delivered in 20 hrs or 1 wk of dedicated teaching time. Factors that impede the provision of undergraduate intensive care teaching are a lack of staff, funding, and dedicated teaching time. PMID- 12545033 TI - Fulminant hepatic failure secondary to acetaminophen poisoning: a systematic review and meta-analysis of prognostic criteria determining the need for liver transplantation. AB - OBJECTIVES: To summarize and compare different prognostic criteria used to determine need for liver transplantation in patients with fulminant hepatic failure secondary to acetaminophen poisoning. DATA SOURCES: Studies published in the literature that investigated criteria for hepatic transplantation secondary to acetaminophen-induced liver failure as identified by a preestablished MEDLINE strategy (1966 through October 2001). STUDY SELECTION: Studies were included if 2 x 2 tables could be reconstructed and if they did not assume that patients undergoing transplantation would have eventually died had they not received the transplant. DATA EXTRACTION: Relevant articles were reviewed by two authors independently. Discrepancies or disagreements, if any, on the inclusion or exclusion of studies were resolved by consulting the third author. DATA SYNTHESIS: King's criteria (pH < 7.30 or prothrombin time of >100 secs plus creatinine of >300 micromol/L plus encephalopathy grade of > or =3) were evaluated in nine studies, pH < 7.30 in four, prothrombin time of >100 secs in three, prothrombin time of >100 secs plus creatinine of >300 micromol/L plus encephalopathy grade of > or =3 in three, creatinine of >300 micromol/L in two, and one each for increase in prothrombin time day 4, factor V of <10%, Acute Physiology and Chronic Health Evaluation (APACHE) II score of >15, and Gc globulin of <100 mg/L. King's criteria were more sensitive than pH: 69% (95% confidence interval, 63-75) vs. 57% (95% confidence interval, 44-68). Their specificities were, however, comparable: 92% (95% confidence interval, 81-97) vs. 89% (95% confidence interval, 62-97). APACHE II score of >15 had the highest positive likelihood ratio (16.4) and the lowest negative likelihood ratio (0.19) but was evaluated in only one study. The accuracy measures of all other criteria were lower than that of King's criteria or pH < 7.30. CONCLUSIONS: Presently, available criteria are not very sensitive and may miss patients requiring transplantation. Future studies should further evaluate the efficacy of the APACHE II criteria. PMID- 12545034 TI - A price for cost-effectiveness: implications for recombinant human activated protein C (rhAPC). PMID- 12545035 TI - Long-term cost effectiveness of drotrecogin alfa (activated): an unanswered question. PMID- 12545036 TI - Targeting the coagulation cascade in sepsis: did we find the "magic bullet"? PMID- 12545037 TI - Crystal ball for acute lung injury prognosis: filled with surfactant? PMID- 12545038 TI - Long-stay patients: are there any long-term solutions? PMID- 12545039 TI - Another definite candidate gene for genetic predisposition of sepsis: interleukin 10. PMID- 12545040 TI - Optimal method of postpyloric tube placement still unclear. PMID- 12545041 TI - Fishing in the gene pool. PMID- 12545042 TI - Low tidal volume ventilation: a PEEP at the mechanisms of derecruitment. PMID- 12545043 TI - Consider saying yes. PMID- 12545044 TI - Defining adrenal insufficiency in septic shock. PMID- 12545045 TI - Hemoadsorption therapy for sepsis syndromes. PMID- 12545046 TI - Endotoxemic myocardial depression: a novel inducible nitric oxide synthase suppressant effect of albumin. PMID- 12545047 TI - Patient with a sudden drop in blood pressure. PMID- 12545048 TI - Major benefit of small tidal volumes during high-frequency ventilation. PMID- 12545049 TI - Dismutating the superoxide ion in sepsis. PMID- 12545050 TI - Rate of change in brain tissue PO2: a novel index of cerebral pressure autoregulation. PMID- 12545051 TI - Cardioversion in the new advanced cardiac life support guidelines. PMID- 12545052 TI - Selective decontamination of the digestive tract: use of the correct antibiotics is crucial. PMID- 12545053 TI - Low-frequency/high-frequency ratio and functional outcome. PMID- 12545054 TI - Critical care excellence. PMID- 12545055 TI - Changes of methodology in clinical trials of sepsis over time. PMID- 12545056 TI - Dilutional acidosis: an endless story of confusion. PMID- 12545073 TI - Candidate HIV/AIDS vaccines: lessons learned from the World's first phase III efficacy trials. PMID- 12545074 TI - Live attenuated, nef-deleted SIV is pathogenic in most adult macaques after prolonged observation. AB - OBJECTIVE: A live attenuated SIV vaccine strain, termed SIVmac239Delta3 and containing large deletions in, and the negative regulatory element, was previously shown to cause AIDS mostly in monkeys vaccinated as infants. In the present study, we demonstrate that SIVmac239Delta3 is pathogenic in most vaccinated adult monkeys, given enough time. METHODS: Eleven rhesus macaques vaccinated as adults with SIVmac239Delta3 were followed for extended periods (up to 6.8 years). RESULTS: We found signs of immune dysregulation in all 11 adult vaccinees. All animals developed persistently inverted CD4 : CD8 T-cell ratios, seven (64%) had persistent recurrent viremia, and six (55%) had decreased CD4 T cell counts (< 500 x 10 cells/l). Further signs included low CD4CD29 lymphocyte subsets, loss of anti-Gag antibodies, anemia, thrombocytopenia, wasting, and opportunistic infections. Two adult vaccinees (18%) subsequently developed AIDS. Development of chronic, recurrent viremia with plasma viral RNA loads > or = 10 copies/ml and cytoviremia was a poor prognostic sign. CONCLUSION: Our data demonstrate that with time, a live attenuated, multiply deleted SIV vaccine can cause immune dysregulation in most vaccine recipients, even in initially immune competent, healthy adults. Immune dysfunction can progress to full AIDS. However, pathogenic effects became evident only several years after vaccination. Thus, mass vaccination of humans with similarly constructed live attenuated HIV vaccines, recently suggested for countries with high HIV-1 transmission rates, seems contraindicated. PMID- 12545075 TI - Latent HIV-1 reactivation in transgenic mice requires cell cycle -dependent demethylation of CREB/ATF sites in the LTR. AB - OBJECTIVE: We previously produced a line of transgenic mice that carried the HIV 1 genome deficient in the gene. Although the HIV-1 genome in the lymphocytes was dormant under normal physiological conditions, it could be reactivated by lipopolysaccharide (LPS) administration via induction of interleukin-1alpha/beta and tumour necrosis factor-alpha. In this report, we analysed further the reactivation mechanism of the latent HIV-1 using this transgenic mouse model. DESIGN: and methods: Possible involvement of CpG methylation in HIV-1 latency was examined by treating transgenic lymphocytes with a demethylating agent, 5' azacytidine. CpG methylation in the HIV-1 long terminal repeat (LTR) was analysed using the bisulfite genomic sequencing method. As previous studies suggested that CpG demethylation depended on the cell cycle progression, we analysed the relation between cell cycle progression and LPS-induced reactivation of HIV-1 by labelling lymphocytes with an intracellular fluorescein, carboxyfluorescein diacetate succinimidyl ester. RESULTS: We found that 5'-azacytidine enhanced HIV 1 expression ninefold compared to treatment with LPS alone. Furthermore, HIV-1 p24 induction by LPS was observed only in cells that had undergone cell division, while induction was prevented in cells in which cell cycle progression was blocked either by mimosine, aphidicolin, or nocodazole. LPS-induced HIV-1 reactivation was associated with demethylation of two CpG sites located in the CREB/ATF binding sites in the HIV-1 LTR in a cell cycle-dependent manner. CONCLUSIONS: These observations indicate that cell cycle progression-dependent demethylation of the CREB/ATF sites in the LTR is crucial for the reactivation of latent HIV-1 genome in transgenic mice. PMID- 12545076 TI - Broad spectrum inhibition of HIV-1 infection by sulfated K5 Escherichia coli polysaccharide derivatives. AB - OBJECTIVE: HIV-1 entry into CD4 cells represents a main target for developing novel antiretroviral agents and microbicides. DESIGN: Sulfated derivatives of the K5 polysaccharide have a backbone structure resembling the heparin precursor, but are devoid of the anticoagulant activity. The derivatives were chemically sulfated in the N position after N-deacetylation, in the O position, or in both sites. METHODS: HIV replication in human T cell blasts, monocyte-derived macrophages and cell lines was studied in the presence of sulfated K5 derivatives. RESULTS: O-sulfated [K5-OS(H)] and N,O-sulfated [K5-N,OS(H)] K5 derivatives with high degree of sulfation inhibited the replication of an HIV strain using CXCR4 as entry co-receptor (X4 virus) in both cell lines and T-cell blasts. K5 derivatives also strongly inhibited the multiplication of CCR5 dependent HIV (R5 virus) in cell lines, T-cell blasts and primary monocyte derived macrophages. Their 50% inhibitory concentration was between 0.07 and 0.46 microM, without evidence of cytotoxicity even at the maximal concentration tested (9 microM). In addition, both K5-N,OS(H) and K5-OS(H) potently inhibited the replication of several primary HIV-1 isolates in T-cell blasts, with K5-N,OS(H) being more active than K5-OS(H) on dual tropic R5X4 strains. K5 derivatives inhibited the early steps of virion attachment and/or entry. CONCLUSIONS: Because K5 derivatives are unlikely to penetrate into cells they may represent potential topical microbicides for the prevention of sexual HIV-1 transmission. PMID- 12545077 TI - Human Leydig cells are productively infected by some HIV-2 and SIV strains but not by HIV-1. AB - OBJECTIVES: With the use of highly active antiretroviral therapy, the identification of HIV reservoirs within the body has become an important issue. However, the testis has been largely ignored despite representing a pharmacologic sanctuary which could act as a viral reservoir. DESIGN: Because alterations in testosterone production have frequently been reported in HIV-infected individuals, we investigated whether the testosterone-producing Leydig cells could become directly infected by HIV-1, HIV-2 or SIV. METHODS: Purified Leydig cells were infected with a panel of HIV-1, HIV-2 and SIV strains and examined for expression of HIV/SIV receptors. Additionally, the impact of CD4 transduction on Leydig cell infection was determined. RESULTS: Leydig cells were unable to support productive infection of the seven HIV-1 isolates tested. No CD4, CXCR4 or CCR5 expression was evident on the surface of Leydig cells and transduction with a CD4 expressing adenovirus did not induce HIV-1 infection. In contrast, some primary and laboratory adapted CD4-independent HIV-2 and SIV strains were able to enter and replicate productively in Leydig cells. CONCLUSIONS: Our results suggest that Leydig cells do not represent a target for HIV-1 infection within the testis. In contrast, Leydig cells support HIV-2 and SIV infection and thus represent a potential target for infection. Receptor use and significance of HIV 2/SIV infection of Leydig cells remain to be determined. PMID- 12545078 TI - Differentiating hyperlipidaemia associated with antiretroviral therapy. AB - BACKGROUND: Hyperlipidaemia associated with antiretroviral treatment has led to concerns for an increased cardiovascular risk in HIV-infected patients. OBJECTIVE: To assess this cardiovascular risk by comparing the lipoprotein pattern of antiretroviral-treated and untreated HIV-positive patients with patients with familial combined hyperlipidaemia (high cardiovascular risk) or familial hypertriglyceridaemia (low cardiovascular risk). METHODS: Fasting serum samples were drawn from consecutive patients with HIV infection or lipoprotein disorders. Total cholesterol, low density lipoprotein (LDL) cholesterol, high density lipoprotein (HDL) cholesterol, triglycerides, apolipoprotein A1 and B were determined in serum. Very low density lipoprotein (VLDL) was prepared by ultracentrifugation and analysed for cholesterol, triglycerides and apolipoprotein B. RESULTS: Lipoprotein disorders were found in 114/187 HIV positive patients (61%). Of these, according to the Fredrickson classification, 10% were type IIa (elevated LDL-cholesterol), 14% type IIb (elevated LDL- and VLDL-cholesterol) and 76% were type IV (elevated VLDL-cholesterol). VLDL composition was analysed in 34 HIV-positive patients with type IV hyperlipidaemia. The ratio of VLDL-triglycerides to VLDL-apolipoprotein B in these patients was 16.2 +/- 6.0. This ratio was not different from 14 patients with famlial hypertriglyceridaemia (16.9 +/- 6.0; = 0.61), but differed substantially from 10 patients with familial combined hyperlipidaemia (6.8 +/- 1.0; < 0.0001). CONCLUSIONS: In HIV-infected patients with high VLDL, large VLDL particles were found with no increase in number. This pattern resembles familial hypertriglyceridaemia. It is different from familial combined hyperlipidaemia, where an increase in number of small-sized VLDL particles occurs. Further research is needed to assess the contribution of VLDL-associated hypercholesterolaemia in those taking antiretroviral drugs to the cardiovascular risk profile of HIV-positive patients. PMID- 12545079 TI - HIV RNA in plasma rebounds within days during structured treatment interruptions. AB - OBJECTIVE: To evaluate time to viral rebound in patients undergoing repeated structured treatment interruptions (STI). METHOD: Fourteen chronically HIV infected patients enrolled in the Swiss-Spanish Intermittent Treatment Trial (SSITT) underwent frequent blood sampling. Patients underwent four cycles of 2 week STI, followed by 8-week retreatment with the identical antiretroviral treatment (HAART) used before STI. At the fifth cycle, treatment was stopped for a longer period. Before each new STI, plasma viral load (VL) had to reach < 50 copies/ml. VL was measured during day 0 (last day on HAART) and on days 4, 8 and 14 during all five STI. RESULTS: During the first cycle, plasma HIV RNA increased to > 50 copies/ml (range, 67-88) in five patients at day 4, in eight patients (> 100 copies/ml) at day 8 and in 12 patients (> 100 copies/ml) at day 14. Cumulative analysis of the frequency of detectable HIV RNA at days 4, 8 and 14 compared with day 0 for all five cycles revealed nine patients with VL > 50 copies/ml [13 of 54 samples tested (24.1%); = 0.14] at day 4, 11 patients [33 of 58 samples tested (56.9%); < 0.0001] at day 8 and 12 patients [53 of 65 samples tested (81.5%); < 0.0001] at day 14. CONCLUSIONS: Significant viral replication can be induced during 1 week STI, and this may increase the risk of the emergence of drug resistance during long-term cycling. Therefore, short-term cycling strategies such as 1-week-on, 1-week-off treatment, although conceptually intriguing, should still be regarded as investigational and should be restricted to rigorously controlled clinical trials ideally involving patients who have never failed treatment before. PMID- 12545080 TI - Influence of polymorphisms within the CX3CR1 and MDR-1 genes on initial antiretroviral therapy response. AB - OBJECTIVE: Single nucleotide polymorphisms (SNP) in the genes encoding the human CX3CR1 chemokine receptor and the P-glycoprotein multidrug transporter have been associated with accelerated disease progression in untreated individuals and implicated in therapeutic response, respectively. This retrospective study assessed the influence of SNP in the CX3CR1 and MDR-1 genes on initial virological and immunological response in 461 HIV-infected, antiretroviral-naive individuals initiating antiretroviral therapy in British Columbia, Canada. METHODS: CX3CR1 and MDR-1 SNP were determined by PCR amplification of human DNA from plasma, followed by DNA sequencing. Time to virological success [time to HIV plasma viral load (pVL) < or = 500 copies/ml], virological failure (subsequent time to the second of two consecutive pVL > or = 500) and immunological failure (time to the second consecutive CD4 cell count below baseline) were analyzed by Kaplan-Meier methods. RESULTS: Frequencies of CX3CR1 amino acid haplotypes were 249V 280T (0.75), 249I 280M (0.15), and 249I 280T (0.1). Frequencies of MDR-1 nucleotide polymorphisms were 3435C (0.47) and 3435T (0.53). There was no effect detected for SNP in CX3CR1 or MDR-1 on time to virological success, nor of CX3CR1 and MDR-1 SNP on time to virological and immunological failure, respectively ( P > 0.1). There was a trend to earlier virological failure in the MDR-1 3435C/C genotype group ( P = 0.07), and a statistically significant trend to earlier immunological failure in individuals with the CX3CR1 249I polymorphism ( P = 0.02). These remained significant after correcting for baseline age, sex, pVL, CD4 cell count, type of therapy, and adherence ( P < or = 0.05). CONCLUSION: Polymorphisms in MDR-1 and CX3CR1 may be associated with accelerated virological and immunological therapy failure, respectively. PMID- 12545081 TI - Efficacy of a twice-daily antiretroviral regimen containing 100 mg ritonavir/400 mg indinavir in HIV-infected patients. AB - OBJECTIVE: To evaluate the pharmacokinetics, efficacy and tolerability of a low dose boosted indinavir (IDV)/ritonavir (RTV) regimen [100 mg RTV/400 mg IDV twice daily (bid)] in patients previously receiving a standard IDV regimen [800 mg three times a day (tid)]. METHODS: In a prospective, open-label, cross-over trial, patients with plasma HIV RNA < 200 copies/ml receiving an IDV-containing regimen (800 mg tid) were switched to an RTV/IDV (100/400 mg bid)-containing regimen. Minimal and maximal IDV plasma concentrations ( Cmin and Cmax ) were determined before the switch (day 0), at week 2 and week 4 after the switch. The CD4 cell count and plasma HIV RNA were determined at day 0, week 2 and week 4, then every 8 weeks. The primary end-point was the percentage of patients with plasma HIV RNA below 200 copies ml at week 48. RESULTS: Twenty patients were enrolled. At baseline, on IDV 800 mg tid, median IDV Cmin was 194 ng/ml and median IDV Cmax was 8449 ng/ml. On RTV/IDV (100/400 mg), median IDV Cmin increased to 536 ng/ml at week 2 and 475 ng/ml at week 4, while Cmax decreased to 2983 ng/ml at week 2 and 2997 ng/ml at week 4 ( P < 0.001). The median area under the IDV plasma concentration-time curve measured in seven patients was 25 126 ng.h/ml, and the IDV half-life (t1/2 ) was 4.4 h. All patients had plasma HIV RNA remaining < 200 copies/ml at week 48. Tolerability of RTV/IDV was excellent. CONCLUSION: RTV/IDV (100/400 mg bid) yields significantly higher IDV plasma Cmin and lower IDV Cmax values relative to the standard IDV regimen, thereby improving both tolerability and efficacy. PMID- 12545082 TI - Risk factors for Kaposi's sarcoma in men seropositive for both human herpesvirus 8 and human immunodeficiency virus. AB - OBJECTIVE: To identify risk factors for Kaposi's sarcoma (KS) among men seropositive for both human herpesvirus 8 (HHV-8) and HIV. DESIGN: Cross sectional study of 91 HHV-8 seropositive, HIV seropositive men who have sex with men (57 with KS), and 70 controls at lower risk for KS. METHODS: Patients received clinical evaluations. Blood, oral fluids, semen, rectal brush, rectal swab, and urine were collected, and tests for HHV-8 were performed. RESULTS: Men with KS were more likely to have HHV-8 DNA in peripheral blood mononuclear cells (PBMC) than men without KS [35.1 versus 5.9%, odds ratio (OR), 8.6, 95% confidence interval (CI), 1.9-39.9]. The prevalence of HHV-8 DNA in oral fluids was similar for the two groups (37.0 versus 32.4%; OR, 1.2; 95% CI, 0.5-3.0). HHV 8 DNA was rarely detected in specimens of other types from these men, or in any specimens from the 70 controls. Among men with KS, HHV-8 DNA in PBMC was associated with new KS lesions (OR, 4.5; 95% CI, 1.4-14.5), and HHV-8 DNA in oral fluids was associated with oropharyngeal KS lesions (OR, 3.1; 95% CI, 1.0-10.1). Men with high HHV-8 antibody titers were more likely to have KS (OR, 9.6; 95% CI, 1.2-78.2), but were less likely to have new KS lesions (OR, 0.2; 95% CI, 0.0-1.1) or HHV-8 DNA in PBMC (OR, 0.2; 95% CI, 0.0-1.6) or oral fluids (OR, undefined; = 0.001). CONCLUSIONS: In HHV-8- and HIV-seropositive men, HHV-8 DNA is associated with KS. Among men without KS, HHV-8 DNA is most commonly found in oral fluids. High HHV-8 antibody titers may protect against circulating HHV-8 and new KS lesions. PMID- 12545083 TI - Evidence of changes in sexual behaviours among male factory workers in Ethiopia. AB - OBJECTIVE: To assess changes in sexual behaviours among male factory workers in Ethiopia. DESIGN: Open cohort studies in two factories near Addis Ababa. DATA AND METHODS: At intake and biannual follow-up visits, data were collected on sexual behaviours including casual sex, sex with commercial sex workers (CSW), condom use, and history of sexually transmitted diseases (STDs) as indicated by genital discharge and genital ulcer. Health education, HIV testing, and counselling were offered to all participants. RESULTS: Between February 1997 and December 1999, 1124 males were enrolled in the two cohort studies. At intake, the prevalence of casual sex in the past year, sex with CSWs, condom use with the last casual partner, history of genital discharge in the past 5 years, and history of genital ulcer in the past 5 years were 9.7, 43.4, 38.8 (Akaki site only), 10.6 and 2.1%, respectively. At the Wonji site, the intake prevalence of casual sex, sex with CSW, and history of genital discharge decreased significantly by calendar year between 1997 and 1999. At both sites combined, between the first and the fourth follow-up visits, there was a decline in the proportion of males reporting recent casual sex (from 17.5 to 3.5%, < 0.001), sex with CSWs (from 11.2 to 0.75%, < 0.001), and genital discharge (from 2.1 to 0.6%, = 0.004). CONCLUSION: There was a decline over time in risky sexual behaviours reported by cohort participants. Part of this decline occurred independently of cohort interventions. PMID- 12545084 TI - Hormonal contraceptive use and HIV-1 infection in a population-based cohort in Rakai, Uganda. AB - BACKGROUND: Hormonal contraceptives have been associated with increased risk of HIV acquisition. METHODS: The association between hormonal contraception use and HIV acquisition was assessed in a rural community-based cohort in Rakai District, Uganda. A group of 5117 sexually active HIV-negative women were surveyed at 10 month intervals between 1994 and 1999. Information on demographic and sociobehavioral characteristics, use of hormonal contraception (pill and injectable methods), condoms and the number of sexual partners was obtained by home-based interview. HIV incidence rate ratios (IRR) and 95% confidence intervals (CI) associated with hormonal contraception were estimated by multivariate Poisson regression after adjustment for age, condom use, number of sexual partners, marital status, education and history of genital ulcer disease. RESULTS: At one or more interviews, 16.6% of women reported use of hormonal contraceptives and 23.0% reported condom use. HIV incidence was 2.3/100 person years in hormonal contraceptive users compared with 1.5/100 person-years in non hormonal contraceptive users (unadjusted IRR, 1.56; 95% CI, 1.00-2.33). After multivariate adjustment, the IRR associated with hormonal contraceptives was reduced to 0.94 (95% CI, 0.53-1.64). The adjusted IRR was 1.12 (95% CI, 0.48 2.56) with oral contraceptive use and 0.84 (95%CI, 0.41-1.72) with injectable methods. CONCLUSION: Use of hormonal contraception is not associated with HIV acquisition after adjustment for behavioral confounding. PMID- 12545085 TI - Dual infections with HIV-1, HIV-2 and HTLV-I are more common in older women than in men in Guinea-Bissau. AB - OBJECTIVES: To investigate the association between the three human retroviruses, HIV-1, HIV-2 and HTLV-I. DESIGN: Community-based follow-up studies of retrovirus infections in two cohorts. METHODS: A total of 2057 individuals aged 35 years and over were eligible for inclusion. Participants were interviewed and had a blood sample drawn. Samples were analysed for HIV-1, HIV-2 and HTLV infections. Uni- and multivariate analyses that included behavioural and socio-economic factors were performed using logistic regression and Poisson regression models. RESULTS: A total of 1686 individuals participated with a blood sample in the HIV prevalence analyses and 1581 individuals participated in the HTLV-I prevalence analyses. The overall prevalence was 2.1% for HIV-1, 13.5% for HIV-2 and 7.1% for HTLV-I. Comparing the < 45 year age group with 45+ year age groups, the female : male (F : M) prevalence ratio increased with age for all three retroviruses. Dual infections were more common in women than in men. Assuming independent distribution of the viruses, the observed prevalence of dual infections in women was significantly higher than expected, while the prevalence was not increased in men. The prevalence of dual infections increased with age in women, the odds ratio (OR) being 3.4 [95% confidence interval (CI), 1.0-11.3] for any combination of dual infection, while the prevalence decreased with age in men (OR, 0.3; 95% CI, 0.1-2.0) (test of interaction, P = 0.033). Control for behavioural factors did not modify these patterns. CONCLUSIONS: The pattern of increased prevalence among older women could have public health implications; women of older age groups should be regarded as a potential vulnerable group and included in HIV/AIDS prevention programmes. PMID- 12545086 TI - Is HIV sequestered in bone? Possible implications of virological and immunological findings in some HIV-infected patients with bone disease. AB - To determine the prevalence of CD4 lymphocyte levels less than 350 cells/mm3 accompanying viral loads (VL) less than 500 copies/ml (discordant CD4 lymphocyte levels/VL) in HIV-infected men with bone pathologies, we conducted a review of the records of 3512 men. We found discordant CD4 lymphocyte levels/VL in 26 (36.1%) out of 72 with bone pathologies, and in 704 (20.5%) out of 3440 without bone pathologies (P < 0.01), and concluded that HIV sequestered in bone was a possible explanation. PMID- 12545087 TI - Low expression of inhibitory natural killer receptors in CD8 cytotoxic T lymphocytes in long-term non-progressor HIV-1-infected patients. AB - We investigated whether a different pattern of HLA-specific inhibitory natural killer receptor (iNKR) expression on peripheral blood mononuclear cells (PBMC) of long-term non-progressor (LTNP) patients explained their benign course of HIV-1 infection. The surface expression of p70, p140 and CD94/NKG2A in their CD3+CD8+ PBMC was comparable to that of uninfected donors. The lack of iNKR-mediated functional inhibition of HIV-1-specific cytotoxic T lymphocytes in vitro could provide an additional mechanism for the efficient control of virus spread in LTNP patients. PMID- 12545088 TI - Differences in presentation and follow-up between HIV-1 antibody positive teenagers and adults: the need for a more focused approach to care? AB - Our findings show a high rate of loss to follow-up in HIV-positive teenagers compared with adults. Of concern is the fact that this group also have high rates of sexually transmitted infections, unprotected sex and pregnancy. There is an urgent need to examine the reasons for this and adopt strategies to minimize risk taking behaviour and to improve access to appropriate healthcare. PMID- 12545089 TI - Individualizing salvage regimens: the inhibitory quotient (Ctrough/IC50) as predictor of virological response. AB - Increased drug levels could overcome resistance and improve the response to a salvage regimen. We evaluate the inhibitory quotient (IQ, Ctrough/protein-binding corrected IC50) as a predictor of virological response in 52 patients included in two dual protease inhibitor (PI)-based salvage regimens. The HIV-RNA level decrease at 12 weeks was greater in patients who achieved an IQ greater than 1. The IQ could be useful to improve the virological response to a dual PI salvage regimen. PMID- 12545090 TI - No increase in protease resistance and a decrease in reverse transcriptase resistance mutations in primary HIV-1 infection: 1992-2001. AB - Rates of antiretroviral resistance in recently transmitted virus in Sydney, Australia fluctuated over the past decade, influenced by treatment trends. Current rates of drug resistance are not high in historical terms or compared with those reported. Rates of resistance to reverse transcriptase inhibitors peaked in the mid-1990s, fell dramatically with the introduction of combination therapy and appear to have plateaued at 10-15% over the past 3 years. Primary resistance mutations in the protease gene are still rare. PMID- 12545091 TI - Beta-glucuronidase activity in patients infected with HIV. PMID- 12545092 TI - European HIV/AIDS case reporting: assessing past treatment history in individuals who develop AIDS in the era of highly active antiretroviral therapy. PMID- 12545093 TI - Case report: nucleoside analogue-induced lactic acidosis in the third trimester of pregnancy. PMID- 12545094 TI - Central nervous system aneurysms in childhood AIDS. PMID- 12545095 TI - Malaria, HIV and pregnancy. PMID- 12545096 TI - Unexpected similarity between the carboxyl termini of lentivirus and pestivirus envelope glycoproteins. PMID- 12545097 TI - HIV and UN peacekeeping operations. PMID- 12545098 TI - Lack of response of anal intra-epithelial neoplasia to highly active antiretroviral therapy. PMID- 12545099 TI - Notes from the 42nd Interscience Conference on Antimicrobial Agents and Chemotherapy, September 2002. PMID- 12545101 TI - Use of octyl-2-cyanoacrylate in cleft lip repair. AB - Octyl-2-cyanoacrylate (Dermabond; Ethicon, Somerville, NJ) is a synthetic tissue adhesive recently approved for skin closure. This study was designed to assess its effectiveness for use in clefts lip repairs. Sixty-four patients with unilateral, bilateral, or midline cleft lip defects were repaired. The ages at repair ranged from 4 days to 19 months, with an average of 46.5 days. Follow-up ranged from 6 months to 3 years. No complications were found. Several advantages were observed: shorter operative time, formation of a protective barrier, simplified incision care, no need for suture removal, and improved scar outcome. This study supports octyl-2-cyanoacrylate as an alternative to skin sutures in primary cleft lip repair. PMID- 12545102 TI - Characteristics of women with cosmetic breast implants compared with women with other types of cosmetic surgery and population-based controls in Denmark. AB - Herein the authors describe characteristics of women with breast implants compared with women with other types of cosmetic surgery as well as population controls. All women who acquired breast implants from 1977 to 1997 were identified from the files of two private plastic surgery clinics in Denmark. Patient characteristics were obtained through a self-administered questionnaire. The magnitude of differences between patient and control groups was estimated using odds ratios and 95% confidence intervals. Women with breast implants had a significantly lower body mass index and reported a two-fold greater incidence of current smoking compared with women from the general population and compared with women with other cosmetic surgery. Women with implants reported a greater number of full-term pregnancies and were less likely than controls to have had their first birth at age 30 years or older. Women with implants were not more likely than women in either control group to report a history of diseases, including connective tissue diseases, cancer, or depression before their implant surgery. Women with cosmetic breast implants differ from women with other forms of cosmetic surgery and from general population controls with respect to characteristics that may importantly influence health outcomes and that need to be addressed in future breast implant studies. PMID- 12545103 TI - Short-term and long-term results of speech improvement after surgery for velopharyngeal insufficiency with pharyngeal flaps in patients younger and older than 6 years old: 10-year experience. AB - Velopharyngeal insufficiency (VPI) is a well-known cause for hypernasality. To overcome this problem, the authors use a static surgical technique: a cranially or caudally based flap. In 93 patients, the results of this technique on speech (hypernasality, nasal air escape, articulation) and velopharyngeal function were evaluated over a period of at least 1 year. In 53 patients, pharyngoplasty flaps were based caudally. In 40 patients, pharyngoplasty flaps were based cranially. The patients were age 2.5 to 24.5 years, with a mean of 5.5 years (SD: 4 years and 2 months). Improvement was found in almost all patients. The patients who underwent surgery when they were younger than age 6 significantly improved better then the patients who were treated when they were older than age 6. There were no differences in outcome between cranially based and caudally based flaps. There were also no differences between patients with plain VPI and patients with VPI (e.g., Pierre Robin sequence and Shprintzen). PMID- 12545104 TI - Palatal surface area measurement: comparisons among different cleft types. AB - The purpose of this study was to use three-dimensional imaging methods to measure the palatal surface of unrepaired cleft patients. The surface area of the palate was defined and measured on three-dimensional computed tomography images of dental plaster models in four different groups of cleft patients at 3 months of age. There were 30 unilateral complete cleft lips and palates (UCLP), 27 bilateral complete cleft lips and palates (BCLP), 23 isolated cleft palates of incomplete form (CP), and 19 unilateral cleft lips without cleft palates (UCL). These patients were nonsyndromic, unoperated, and without other major deformities. The dental casts were scanned, and the computed tomography data were transferred to an imaging laboratory for processing and reconstruction of three dimensional images. Surface area of the palate was delineated, which was defined as within the alveolar crest and the line connecting both tuberosities. In UCLP and BCLP, the edge of cleft formed the medial boundary of the area for each palatal shelf, and the palatal surface area was the combination of both palatal shelves and the premaxillary area in BCLP group. The surface area was measured. Repeated definition and measurement tasks were performed for calculation of errors. The imaging data management and measurement were performed using the Analyze program (Biomedical Imaging Resource, Mayo Foundation, MN). In addition, linear distances were measured between the canine points on the alveolar crest (line C) and the tuberosity points (line T). The measurements were compared among the different groups. Analysis of variance and multiple comparisons were used for statistical analyses. The results showed that the mean error between repeated area definitions and measurements in this study was 1.86%. The bilateral complete cleft lip and palate (BCLP) and unilateral complete cleft lip and palate (UCLP) groups had significantly smaller palatal surface area than the unilateral cleft lip without cleft palate (UCL) and isolated cleft palate of incomplete form (CP) groups. There was no significant difference between the BCLP and UCLP groups. Line C and line T distances were significantly longer in BCLP and UCLP groups than in UCL and CP groups. The findings suggest that compared with UCL and CP patients, there is an intrinsic tissue deficiency in the palate/maxilla of BCLP and UCLP patients. PMID- 12545105 TI - Influence of periosteum on donor healing after harvesting hard palate mucosa. AB - The authors report the influence of periosteum on healing of palatal defect based on more than 10 years of experience of harvesting hard palate mucosa. Between June of 1991 and May of 2001, the authors harvested 80 hard palate mucosae as graft material for skin and mucosa defects. All grafts were harvested from the center of the hard palate. Patients ranged in age from 10 to 82 years old. Of 80 mucosae, 54 were harvested with periosteum, and periosteum was not retained in the defect bed. The other 26 mucosae were harvested without periosteum, which was therefore retained in the defect bed. The healing time increased depending on the defect size in both groups of patients retaining and not retaining periosteum. There was a significant relationship between the defect size and healing time in both groups (Spearman's rank correlation test, p < 0.0001 in both groups). In the two groups, there was no significant relationship between patient age and healing time in the patients with defect smaller than 1.99 cm or larger than 2.00 cm2. There were no significant differences in the rate of patients with pain and bleeding between the groups retaining and not retaining periosteum. In the group not retaining the periosteum, all 54 patients showed a flat or atrophic smooth surface at more than 6 months after epithelization and had no discomfort. However, 17 patients showed flat or atrophic smooth surface in the group retaining the periosteum and the remaining 9 patients showed hypertrophy at more than 6 months after epithelization, with accompanying discomfort. The rate of the patients with hypertrophy in the group of patients retaining periosteum was significantly high as compared with that in the group of patients not retaining periosteum (p = 0.000013, Fisher's exact test). In 26 patients retaining periosteum, the age of the patients with hypertrophic surface was significantly younger than that of the patients with flat or atrophic surface (p = 0.0010, Welch's -test), and the defect size in the patient with hypertrophic surface was significantly smaller than that of the patients with flat or atrophic surface (p = 0.0028, Welch's t-test). In conclusion, our study demonstrated that the existence of periosteum in the palate donor bed does not contribute to reduced healing time or reduced pain. Rather, retaining the periosteum caused hypertrophy of the donor site, leading to discomfort, especially in young patients with a comparatively small defect. PMID- 12545106 TI - New technique for nipple areola reconstruction: arrow flap and rib cartilage graft for long-lasting nipple projection. AB - The nipple-areola complex is an integral part of breast reconstruction. It serves as an important landmark for the new breast and many techniques have been described for its reconstruction. Historically, nipple projection has been most difficult to achieve and maintain. The authors found that techniques that use isolated soft tissue flaps eventually fail secondary to scar contraction with loss of projection. Since 1994, the authors have used a local skin fat flap shaped as an arrow for the reconstruction of the nipple on 454 breasts. The arrow shape is advantageous because it allows the scar to be broken on closure, minimizing postoperative wound contracture. Within the flap, the authors routinely use a rib cartilage graft, which provides additional support and projection for the skin-soft-tissue envelope. The graft is harvested during the initial breast flap transfer for the purpose of exposing the internal mammary vessels and has been obtained without any additional morbidity. By combining a skin fat flap and rib cartilage graft, the authors have achieved excellent long term projection and a more esthetically pleasing nipple reconstruction. PMID- 12545107 TI - Orbital reconstruction after exenteration: use of a transorbital temporal muscle flap. AB - Orbital exenteration is a disfiguring operation that involves the total removal of the orbital contents with partial or total excision of the eyelids. Common methods of orbit reconstruction include pectoralis musculocutaneous pedicled flap and free tissue transfer. The purpose of this study is to illustrate that the entire temporalis muscle may be used by creating a large window in the lateral orbit, without resection of the lateral orbital rim. Orbital exenteration was performed on four cadavers. A window was created in the lateral orbit using a 4 mm pineapple burr. Three parameters were measured: (1) the distance between the zygomatic arch to the superior aspect of the temporalis muscle; (2) the width of the temporalis muscle; and (3) the length and width of the lateral orbit window. The free edge of the transposed temporal muscle was then sutured to the skin edge around the bony orbit. This procedure was then performed on a 73-year-old man who had undergone right orbital exenteration for ocular melanoma and then postoperative radiation. The dimensions of the bony windows in the cadavers were as follows: mean 3.3 cm (SD +/- 0.19 cm) x 1.9 cm (SD +/- 0.18 cm), n = 4. The dimensions of the temporalis muscle in the cadavers were the following: mean 8.45 cm (SD +/- 0.60 cm) x 10.5 cm (SD +/- 0.33 cm), n = 4. In the patient, the size of the bony window was 3.7 cm x 2.1 cm (n = 1), and the dimensions of the temporalis muscle were 8.1 cm x 10.2 cm (n = 1). The patient recovered well without complication, with a well-healed skin graft over the top of the muscle flap. An adequate bony window can be made to allow transfer of the entire temporalis muscle for orbital reconstruction without resecting the lateral orbital rim or entering the middle cranial fossa. This option is a good alternative to the other commonly performed methods of orbital reconstruction because of its completion in one operative stage, short operative time, and minimal donor site morbidity. PMID- 12545108 TI - Management of lower lip cancer: a retrospective analysis of 118 patients and review of the literature. AB - In this current study, the clinical data and postoperative follow-up findings of 118 patients with a primary lower lip carcinoma who were treated between 1983 and 1999 in the Department of Plastic and Reconstructive Surgery are presented. Medical records were reviewed retrospectively and data were collected concerning age, gender, followup period, location of lesion on the lip, cervical metastasis at presentation, preoperative biopsy results, histological grade, initial treatment, reconstruction type, pathological outcome, local recurrence, regional lymph node metastasis, treatment of local recurrence and regional lymph node metastasis, and postoperative treatment. The prognostic value of clinical stages in relation with recurrence and mortality from disease was investigated. The overall rate of recurrence was calculated as being 39.8%, and the determinate survival rate was found to be 72.9% at 5-year follow-up. The data concerning the above-mentioned parameters, together with risk factors that might play a role in the development of lip cancer, are discussed in light of the current literature. PMID- 12545109 TI - Effect of hyperbaric oxygen therapy on the tubed pedicle flap survival in a rat model. AB - The effect of hyperbaric oxygen therapy (HBO) on survival of the distal skin paddle with early pedicle division was studied in a rat tubed pedicle flap model. In part 1, tubed pedicle skin flaps were created on the backs of 14 rats. The pedicle of each skin flap was divided at intervals of 3, 4, 5, and 7 days, and the survival area of each distal skin paddle was measured 5 days after each pedicle was divided. The results showed that the percentages of survival were 0%, 29.9%, 89.9%, and 100% at 3, 4, 5, and 7 days, respectively. In part 2, the tube flaps were created on the backs of 28 rats. The tubed pedicles were divided 4 days after creation. In the experimental group 1 (n = 9), the rats received HBO treatment at 2 atm of pressure twice daily for 3 consecutive days before pedicle division. In the experimental group 2 (n = 9), the rats received HBO treatment after pedicle division twice daily for 3 consecutive days. In the control group (n = 10), the rats received no HBO treatment. The results showed that the mean survival area of the distal skin paddle at 5 days after pedicle division for the groups treated with HBO before and after pedicle division was 6.31 +/- 2.69 cm2 (69.19%) and 6.12 +/- 1.52 cm2 (71.22%), respectively. These survival rates were of statistical significance as compared with the survival rates in the control group of 2.70 +/- 1.89 cm2 (27.9%). These results demonstrate that the administration of HBO therapy to the animal with a tubed pedicle flap can increase flap survival with early division of the tubed pedicle. PMID- 12545110 TI - Reactive thrombocytosis without endothelial damage does not affect the microvascular anastomotic patency. AB - There is still controversy about the correlation of thrombocytosis and thrombosis complication. Using a rodent splenectomy-induced thrombocytosis model and a thrombogenic endothelial damage model (inverted suture resulting in an intraluminal thrombogenic adventitia of divided femoral artery), the authors investigated whether reactive thrombocytosis with or without endothelial damage contributes to the patency of microvascular anastomosis. Four experimental groups were evaluated in this study: 1) sham operation without thrombogenic anastomosis after femoral artery division; 2) sham operation with thrombogenic anastomosis; 3) thrombocytosis alone without thrombogenic anastomosis; 4) thrombocytosis with thrombogenic anastomosis (each subgroup n = 10, total N = 40). Vascular patency was assessed after immediate operation and on the seventh day postoperatively. Platelet counts and platelet activation (CD62P) were studied in correlation to microvascular patency. In rats without thrombogenic anastomosis groups, there were no significant differences in CD62P expression on platelets (p = 0.09), the patency rates (p = 0.561), or perfusion units (p = 0.746) before and after arterial reanastomosis between rats with and without thrombocytosis, respectively. However, the thrombogenic anastomosis of femoral artery in thrombocytosis and control groups showed significantly increased CD62P expression (p < 0.05), decreased the perfusion unit (p < 0.05), and patency rate (p < 0.001), compared with rats without thrombogenic anastomosis of femoral artery in both groups. In summary, this study demonstrates that microvascular anastomosis can be performed safely with reactive thrombocytosis alone without thrombogenic anastomosis. Meticulous microvascular anastomosis without triggering platelet activation is the most important factor to prevent thrombosed vessels in microsurgical anastomosis. PMID- 12545111 TI - Age-dependent changes of postischemic reperfusion in rat skin. AB - Ischemia-reperfusion plays a certain role in causing skin damage associated with pressure sores. In this study, changes in cutaneous hemodynamics during reperfusion were investigated in young and older rats. After cessation of 1-hour or 2-hour ischemia, the skin blood flow increased transiently (postischemic hyperemia) and quickly returned to the baseline in young and older rats. After 4 hour ischemia, however, the postischemic hyperemia was reduced in both groups, and the skin blood flow decreased below the baseline for a few hours in older rats. The skin blood flow tolerated well the repeated exposures to 1-hour ischemia in both groups. In 2-hour ischemia experiments, the postischemic hyperemia was preserved after the second ischemic period in young rats but not in older rats. These results suggest that the tolerance of skin microcirculation to ischemia-reperfusion may decrease with increasing age. PMID- 12545112 TI - Effect of mitomycin C on keloid fibroblasts: an in vitro study. AB - Keloids are the result of aberrant wound healing of human skin after dermal injury. Therapeutic options include excision followed by radiation therapy, steroid injection, and compression with silicone sheets among others. Local invasion and recurrence after excision has provoked interest in treating keloids as neoplasms. The purpose of this study was to determine the effect of mitomycin C (MMC) on keloid fibroblasts. Keloid fibroblasts obtained from five different patients were exposed to MMC. A control group of normal and keloid cells was treated with phosphate buffered saline only. Contrast microscopy showed a decrease of fibroblast density during the 3 weeks after exposure for normal and keloid fibroblasts relative to untreated fibroblasts. This was confirmed by total cell counts ( = 0.1) and measurement of DNA synthesis. By the third week, there was a recovery in DNA synthesis and increased cell count for some of the treated fibroblasts. We concluded that at an appropriate concentration, MMC shows proliferation of keloid fibroblasts in vitro for a period of 3 weeks. This agent may be considered in clinical trials after surgical excision of keloids. PMID- 12545113 TI - Resurfacing of a totally degloved hand using thin perforator-based cutaneous free flaps. AB - Resurfacing after a total degloving injury to the hand is one of the most difficult management problems in hand surgery. Although there are many methods of managing this type of injury that preserve functions and lessen deformities, none provides a satisfactory solution to this problem. The authors resurfaced a totally degloved hand using extremely thin and broad perforator-based cutaneous free flaps, and the donor defects were covered with split-thickness skin grafts. The postoperative course was uneventful, the flaps survived completely, and the grafts took without loss. Several minor operations, including interdigitation, defatting, and the formation of palmar and digital creases, were required to obtain the final appearance and function of the hand. Eighteen months after the initial operation, the patient could pick up a bean with a pair of chopsticks. Sensation was satisfactory in the palm 20 months after the initial operation, as evidenced by 10 mm of static two-point discrimination. To reconstruct a total and complete skin defect of the hand, the authors recommend that thin perforator based cutaneous free flaps be an initial consideration. PMID- 12545114 TI - Correction of auricular deformity caused by high ear-piercing: case report. AB - In recent years, high ear-piercing and wearing earrings in the cartilaginous region of the auricle have become popular. The frequent occurrence of subsequent complications also has been pointed out. The present case is of a 17-year-old girl with an auricular deformity caused by infections occurring after piercing in the scapha. The authors performed an auriculoplasty by excision of the deformed cartilage and autologous cartilage graft from the ipsilateral concha. At present, 1 year and 5 months after the operation, no sign of recontracture is noted, and the auricle remains in a gratifying shape. PMID- 12545115 TI - Vulvo-perineal reconstruction: medial thigh septo-fascio-cutaneous island flap. AB - From 1996 through 2001, the authors performed 26 vulvo-perineal reconstructions after vulvar cancer resection. A unilateral or bilateral medial thigh septo fascio-cutaneous island flap was used in all cases. The vascular supply to this flap is guaranteed by the underlying fascial plexus that arises from the convergence of three longitudinal rows of vessels (anterior, medial, and posterior). The flap is fashioned in a triangle shape and incorporates the underlying muscular fascia, sparing the intermuscular septal perforators. It is advanced in a V-Y fashion and is easily adapted and secured to the recipient site. Competent urethral and anal meati are reconstructed with multiple intercalated flaps. Scarring at the donor site is limited and acceptable. The nerve supply guarantees acceptable sensation at the reconstructed area. The medial thigh septo-fascio-cutaneous flap allows a valid one-stage vulvo-perineal reconstruction with a simple surgical execution process after superficial soft tissue resections. PMID- 12545116 TI - Perforator flaps: evolution, classification, and applications. AB - In this article, the authors review the literature regarding perforator flaps. Musculocutaneous perforator flaps have evolved from musculocutaneous flaps and offer several distinct advantages. By sparing muscle tissue, thus reducing donor site morbidity and functional loss, perforator flaps are indicated for a number of clinical problems. The versatility of the perforator flap makes it ideal for the reconstruction of three-dimensional defects such as breast reconstruction or as a thin flap for resurfacing shallow wounds when bulk is considered a disadvantage. The authors review the historical development of the perforator flap and discuss the advantages and disadvantages of perforator flaps compared with free and pedicled musculocutaneous flaps. The nomenclature traditionally used for perforator flaps is confusing and lacks a standardized anatomic basis. The authors present a method to describe all perforator flaps according to their artery of origin. PMID- 12545118 TI - Re: Epineural sleeve neurorrhaphy. PMID- 12545119 TI - Re: Epineural sleeve neurorrhaphy. PMID- 12545120 TI - Re: V-Y advancement flap. PMID- 12545121 TI - Re: Perforator-based forearm and hand adipofascial flaps for the coverage of difficult dorsal hand wounds. PMID- 12545122 TI - Intermaxillary fixation using drill-free screws. PMID- 12545123 TI - Reply: Cutaneous anthrax of the eyelid. PMID- 12545124 TI - Honey cream. PMID- 12545125 TI - Reconstruction of an oral cavity defect resulting from verrucous carcinoma with a prefabricated parietal galeal flap. PMID- 12545126 TI - Use of the real tumescent technique: anesthesia in the surgical management of lip cancer. PMID- 12545127 TI - Fasciocutaneous flap to treat verneuil disease (hidradenitis suppurativa) in the axillary region. PMID- 12545128 TI - Bowenoid papulosis. PMID- 12545130 TI - [Specific complications after lung transplantation for cystic fibrosis]. AB - Complications observed after lung transplantation are well described for the different indications. Certain complications are specifically observed in patients with cystic fibrosis who have reached the stage of major respiratory failure. The most specific complications result from digestive tract manifestations of the disease: malabsorption of lipophilic drugs, osteoporosis, diabetes. These patients also have a risk of infection: carriage of resistant strains, sinus colonization, poor immunization against certain viruses. They are particularly susceptible to post-transplantation lymphoproliferative disorder and neurological drug toxicity. Finally, in certain emergency situations, grafts from living donors may create specific problems. Despite these complications, lung transplantation remains the most effective treatment for respiratory failure due to cystic fibrosis. PMID- 12545131 TI - [Management of patients infected by the human immunodeficiency virus (HIV). Report of an expert group]. PMID- 12545132 TI - [Prevalence of asthma in subjects aged 15-64 years in Bobo-Dioulasso (Burkina Faso) in 1998]. AB - The purpose of this study was to estimate the prevalence of asthma in the city of Bobo-Dioulasso (Burkina Faso) and detail the epidemiological pattern. A screening questionnaire was administered to a representative sample of the city population: 808 subjects aged 15 to 64 years. Persons presenting signs suggestive of asthma were asked to attend consultations at the lung disease unit of the Souro-Sanou Hospital at Bobo-Dioulasso for physical examination. The prevalence of asthma was 9.6%. In the asthma population, 80.8% were aged 15-45 years, with no difference between men and women. Signs of asthma predominated during the evening and night (57.5%). Both benign and intermittent courses were observed. There was a peak during the cold dry season (44% of the acute episodes). Asthma was found in subjects from all ethnic origins but with different distributions. Asthma was found to be an important public health problem in Bobo-Dioulasso. Further studies should be conducted throughout the country to obtain more precise information on asthma and better detail the different aspects in order to develop prevention measures. PMID- 12545133 TI - [Askin tumor: two cases]. AB - Askin tumor is uncommon, usually observed in young subjects. We report two cases of Askin tumor observed in patients aged 15 and 27 years who were hospitalized for a painful tumefaction of the chest wall with pleural involvement in one patient. We reviewed progress in patient management which has resulted from better understanding of the natural history of Askin tumor and earlier diagnosis due to advances in imaging and immunohistochemistry techniques as well as cytogenetics. A number of points remain controversial. PMID- 12545134 TI - [Sarcoid-like necrotizing granulomatosis. A case report]. AB - Necrotizing sarcoid granulomatosis is a rare granulomatous pulmonary angeitis that was first described by Liebow in 1973. We report the case of a 36-year-old woman who was admitted for exploration of chest pain, cough and fever at 39 degrees which had progressed for more than 2 months. Chest X-ray showed several inconstant pulmonary opacities. The physical examination, the radiographic and endoscopic aspects were non-specific. The diagnosis was made only by open lung biopsy which showed histological findings of necrotizing sarcoid granulomatosis. The opacities disappeared with no recurrence spontaneously at one year. PMID- 12545135 TI - [Descending necrotizing mediastinitis: a diagnosis not to miss]. AB - Descending necrotizing mediastinitis is a severe disease which occurs after a mild otorhinolaryngologic or dental infection. The diagnosis must be established rapidly with the help of clinical and computed tomography of the neck and chest data. Treatment is based on antibiotics, surgery and hyperbaric oxygen. The outcome is poor with high mortality. We report a case of septicemia complicating descending necrotizing mediastinitis after dental infection. PMID- 12545136 TI - [New dietary guidelines before FDG-PET, or how to simply improve validity]. PMID- 12545137 TI - Clinical pharmacology & therapeutics--an update. PMID- 12545138 TI - Drug labeling should be kept current. PMID- 12545139 TI - A rational approach for selection of optimal covariate-based dosing strategies. AB - BACKGROUND: At present, there is no rational approach for choosing a dosing strategy for individualization based on a covariate. An approach to use in establishment of an a priori dosing strategy for individualization is presented. Factors influencing the choice of such a dosing strategy are identified. METHODS: The approach requires definition of the following: target variable, seriousness of deviations from the target (ie, risk function), population model, covariate distributions, and constraints. Minimizing the total risk yields an optimal dosing strategy, estimated as dose sizes for different subpopulations and covariate cutoff values at which doses are increased or decreased. The method was illustrated with the use of simulated and real drug examples for the situation in which clearance is related to creatinine clearance. RESULTS: The estimated optimal cutoff(s) paralleled the median creatinine clearance in the population. The extent of variability in clearance explained by creatinine clearance was the main factor influencing the optimal ratios between adjacent dose sizes. An optimal dosing strategy was possible to estimate for the real drug. CONCLUSIONS: The method is simple to perform, although one difficulty lies in defining the target variable and risk function. Our results imply that commonly used constraints in dosing strategies based on renal function (ie, dose ratio of 2 and predetermined cutoffs) are nonoptimal in the sense we propose. Because an optimal dosing strategy may not be practical to use, the therapeutic cost that would result with any constraint can be assessed by comparison of the outcome after the desired and the optimal strategy. PMID- 12545140 TI - Population pharmacokinetics and effects of efavirenz in patients with human immunodeficiency virus infection. AB - OBJECTIVE: The reverse transcriptase inhibitor efavirenz is currently used at a fixed dose of 600 mg/d. However, dosage individualization based on plasma concentration monitoring might be indicated. This study aimed to assess the efavirenz pharmacokinetic profile and interpatient versus intrapatient variability in patients who are positive for human immunodeficiency virus, to explore the relationship between drug exposure, efficacy, and central nervous system toxicity and to build up a Bayesian approach for dosage adaptation. METHODS: The population pharmacokinetic analysis was performed by use of NONMEM based on plasma samples from a cohort of unselected patients receiving efavirenz. With the use of a 1-compartment model with first-order absorption, the influence of demographic and clinical characteristics on oral clearance and oral volume of distribution was examined. The average drug exposure during 1 dosing interval was estimated for each patient and correlated with markers of efficacy and toxicity. The population kinetic parameters and the variabilities were integrated into a Bayesian equation for dosage adaptation based on a single plasma sample. RESULTS: Data from 235 patients with a total of 719 efavirenz concentrations were collected. Oral clearance was 9.4 L/h, oral volume of distribution was 252 L, and the absorption rate constant was 0.3 h(-1). Neither the demographic covariates evaluated nor the comedications showed a clinically significant influence on efavirenz pharmacokinetics. A large interpatient variability was found to affect efavirenz relative bioavailability (coefficient of variation, 54.6%), whereas the intrapatient variability was small (coefficient of variation, 26%). An inverse correlation between average drug exposure and viral load and a trend with central nervous system toxicity were detected. This enabled the derivation of a dosing adaptation strategy suitable to bring the average concentration into a therapeutic target from 1000 to 4000 microg/L to optimize viral load suppression and to minimize central nervous system toxicity. CONCLUSIONS: The high interpatient and low intrapatient variability values, as well as the potential relationship with markers of efficacy and toxicity, support the therapeutic drug monitoring of efavirenz. However, further evaluation is needed before individualization of an efavirenz dosage regimen based on routine drug level monitoring should be recommended for optimal patient management. PMID- 12545141 TI - Population analyses of sustained-release verapamil in patients: effects of sex, race, and smoking. AB - OBJECTIVE: Our objective was to determine the effects of age, sex, and sustained release formulation on apparent oral clearance of sustained-release racemic verapamil in patient populations. METHODS: Population pharmacokinetic analyses were performed on data from 186 patients with hypertension, coronary artery disease, or supraventricular arrhythmias who were receiving long-term sustained release oral racemic verapamil (Covera SR in 105 patients, Calan SR in 67 patients, and other formulations in 14 patients; mean +/- SD dose, 280 +/- 139 mg) for clinical care or as a part of phase III efficacy studies. Of those 186 patients, 135 were men (age, 63 +/- 12 years; ideal body weight, 70.7 +/- 6.6 kg) and 51 were women (age, 60 +/- 17 years; ideal body weight, 53.7 +/- 7.2 kg). Verapamil was measured by HPLC, and population analyses were performed by use of NONMEM software. Sex, age, and formulation were the covariates considered in the population model building. Subgroup analyses of race, smoking, and alcohol consumption were also performed. Significance of covariates was determined by likelihood ratio tests. RESULTS: Sex significantly affected steady-state clearance of oral sustained-release racemic verapamil. Apparent oral clearance of sustained-release verapamil was 23.8 +/- 2.3 mL/min per kilogram in women compared with 18.6 +/- 3.4 mL/min per kilogram in men. Clearance estimates were faster in black subjects compared with white subjects, as well as in smokers compared with nonsmokers. Effects of age, formulation, and alcohol consumption were not detected. CONCLUSIONS: In middle-aged and older patients, apparent oral clearance of sustained-release racemic verapamil was affected by sex (faster in women compared with men), race (faster in black subjects compared with white subjects), and smoking (faster in smokers compared with nonsmokers) but not by age, alcohol, or formulation. PMID- 12545142 TI - Coordinate induction of both cytochrome P4503A and MDR1 by St John's wort in healthy subjects. AB - BACKGROUND: Many drugs are cosubstrates of cytochrome P450 (CYP) 3A and MDR1; furthermore, their disposition is markedly affected by pretreatment with inducing agents, including St John's wort. Such drug interactions reflect induction of both proteins through a common mechanism involving the steroid X receptor/pregnane X receptor. However, the relative contributions of enhanced metabolism and efflux transport to the overall induction process are unknown. METHODS: The effects of 12 days' pretreatment with St John's wort on the disposition of selected in vivo probe drugs were determined in 21 young healthy subjects. Midazolam after oral and intravenous administration was used to assess CYP3A activity in both the intestinal epithelium and the liver, whereas the disposition of fexofenadine after an oral dose was assumed to be a measure of MDR1 function, and the oral plasma concentration-time profile of cyclosporine (INN, ciclosporin) was considered to reflect both CYP3A and MDR1 activities. RESULTS: St John's wort markedly affected the plasma concentration-time profiles of all of the drugs, with associated increases in their clearance. With midazolam, the enhancement was considerably less after intravenous administration (approximately 1.5-fold) than after oral administration (approximately 2.7-fold), and estimated intestinal and hepatic extraction ratios were higher by approximately 1.2- to 1.4-fold. By contrast, the oral clearances of fexofenadine and cyclosporine were equally increased by approximately 1.6-fold and 1.9-fold, respectively; these changes were both statistically less than for midazolam's oral clearance and greater than its estimated intestinal extraction. CONCLUSIONS: Although the disposition of all 3 drugs was altered by St John's wort, the extent of induction measured by oral clearance was different with CYP3A activity (midazolam), apparently increasing more than MDR1 function (fexofenadine), whereas with cyclosporine the change in oral clearance appeared to be more closely associated with the increase in MDR1 rather than CYP3A, despite the fact that both proteins are importantly involved in its disposition. These discordances indicate that, although a common molecular mechanism may be involved, the quantitative aspects of induction are complex and depend on the particular drug and the relative contributions of CYP3A and MDR1 in its disposition. PMID- 12545143 TI - Dipyridamole enhances digoxin bioavailability via P-glycoprotein inhibition. AB - BACKGROUND: On the basis of in vitro studies indicating that dipyridamole is an inhibitor for the MDR1 efflux membrane transporter P-glycoprotein, we postulated that dipyridamole could increase the bioavailability of digoxin, a P-glycoprotein substrate. OBJECTIVES: The main objective was to determine whether dipyridamole alters the bioavailability of digoxin. The secondary objective was to determine whether the magnitude of the pharmacokinetic interaction was influenced by MDR1 genetic polymorphism in exon 26 (C3435T). MATERIAL AND METHODS: (1) The effect of dipyridamole on in vitro P-glycoprotein-mediated, polarized transport of tritium labeled digoxin was investigated in Caco-2 cell monolayers. (2) Twelve healthy volunteers participated in this open, randomized, 2-period crossover study, in which the effects of dipyridamole (300 mg/d for 3 days) versus placebo on the pharmacokinetics of a single oral dose of digoxin (0.5 mg) were compared. MDR1 genotyping (exon 26, C3435T) was determined before the study to include 6 homozygous CC and 6 homozygous TT subjects. RESULTS: Dipyridamole inhibited [(3)H]digoxin transport in Caco-2 cells with a 50% inhibitory concentration value of 1.5 +/- 1.5 micromol/L. We observed a 20% and 13% increase in digoxin area under the plasma concentration-time curve (AUC) from 0 to 4 hours and AUC from 0 to 24 hours (P <.05), respectively, during dipyridamole administration, which was consecutive to an increase in digoxin absorption. Digoxin AUC from 0 to 4 hours and AUC from 0 to 24 hours were significantly higher among subjects harboring the TT compared with the CC MDR1 genotype: 7.5 +/- 1.2 ng x h x mL(-1) versus 6.1 +/- 0.8 ng x h x mL(-1) and 20.2 +/- 2.1 ng x h x mL(-1) versus 16.8 +/- 1.7 ng x h x mL(-1), respectively (P <.05). Digoxin pharmacokinetic modifications during the dipyridamole period were similar in both genotypes. CONCLUSION: Dipyridamole is an in vitro and in vivo P-glycoprotein inhibitor that increases intestinal digoxin absorption and digoxin plasma concentrations. In light of the modest changes in digoxin pharmacokinetics in the presence of dipyridamole, this drug interaction is probably clinically irrelevant. PMID- 12545144 TI - Effects of age and sex on the disposition of retigabine. AB - BACKGROUND: The novel antiepileptic drug retigabine is the first selective M current potassium channel opener for KCNQ2/3 and KCNQ3/5 channels. Retigabine undergoes phase II metabolism (N-glucuronidation, acetylation) exclusively and renal excretion. OBJECTIVE: Our objective was to evaluate the effects of age and sex on the pharmacokinetics of retigabine. METHODS: Healthy young (age range, 18 40 years) and elderly (age range, 66-81 years) white subjects (12 men and 12 women in each group) received a single 200-mg oral dose of retigabine. After dosing, blood was collected over a 72-hour period to determine plasma concentrations of retigabine and its acetylated metabolite, AWD21-360. Pharmacokinetics was compared for age group and sex by ANOVA. RESULTS: In young men, retigabine was rapidly absorbed, with the maximum concentration occurring within 2 hours, and was eliminated with an apparent clearance of 0.67 L x h(-1) x kg(-1) and a mean terminal half-life of 8.5 hours. Subjects were similarly exposed to AWD21-360. Compared with young men, young women had higher retigabine maximum concentration (56%) and exposure (20%) but similar clearance (0.68 L x h( 1) x kg(-1)); these differences were related to differences in body weight. Although maximum concentration was similar in elderly subjects, retigabine elimination was slower (30% lower apparent clearance normalized for weight), resulting in higher exposure (42%) and a longer half-life (30%). Because phase II metabolism is scarcely affected by age, these differences may be related to the known decline of renal function with age. CONCLUSIONS: Although there are no substantial sex-related differences in the disposition of retigabine, a relevant decrease in clearance resulting in higher exposure occurs in elderly patients. The results suggest that decline of renal function with age may account for some of the observed changes. PMID- 12545145 TI - Transfer of metformin into human milk. AB - OBJECTIVES: Our objectives were to determine the milk-to-plasma ratio of metformin in lactating mothers and to estimate infant exposure. METHODS: Two studies were performed. In study 1, 3 nursing mothers taking metformin were studied throughout a dosing interval at steady state. Blood samples were obtained from 2 suckling infants. In study 2, 5 healthy lactating women who volunteered to express milk after weaning were given metformin, 500 mg, at weaning and were studied for up to 72 hours. In both studies, areas under the plasma and milk concentration-time curves were estimated, and the milk-to-plasma concentration ratio based on area under the concentration-time curve analysis was derived. The infant dose was calculated by standard methods. RESULTS: In study 1 the milk-to plasma concentration ratios based on area under the concentration-time curve analysis were 0.37, 0.50, and 0.71. The estimated "doses" of metformin that would be ingested by the breast-fed infants were 0.18%, 0.20%, and 0.21% of the maternal doses, adjusted for weight. In the breast-fed infants, no metformin was detected (n = 2) or adverse effects noted (n = 3). In study 2, the milk-to-plasma concentration ratio based on area under the concentration-time curve analysis was unable to be calculated for 3 subjects because of the unexpected persistence of metformin in milk beyond the study period. For the 2 subjects studied for 72 hours, the milk-to-plasma concentration ratios based on area under the concentration-time curve analysis were 0.27 and 0.47 and the infant doses were 0.11% and 0.25%. The concentration-time profile for metformin in milk in all subjects was unexpectedly flat. CONCLUSIONS: Metformin appears to be "safe" during lactation because of low infant exposure. The unusual concentration-time profile for metformin in milk suggests that the transfer of metformin into milk is not solely dependent on passive diffusion. PMID- 12545146 TI - Effects of nelfinavir and its M8 metabolite on lymphocyte P-glycoprotein activity during antiretroviral therapy. AB - The efflux pump P-glycoprotein decreases drug penetration into cells and tissues. To determine whether nelfinavir or its metabolites inhibit P-glycoprotein in lymphocytes from a healthy volunteer, whole blood cells from human immunodeficiency virus-negative donors were incubated either in human plasma to which nelfinavir or its M8 metabolite were added ex vivo or in plasma from human immunodeficiency virus-positive patients receiving nelfinavir. The 50% P glycoprotein inhibitory concentrations of purified nelfinavir and M8 were 10.9 micromol/L and 29.5 micromol/L, respectively, for CD4(+) T cells and 19.3 micromol/L and >48 micromol/L, respectively, for CD8(+) T cells. Significant inhibitory activity was present in plasma from 27 of 46 patients (59%) receiving nelfinavir. Plasma nelfinavir concentrations correlated with percent inhibition on CD4(+) (rho = 0.85, P <.0001) and CD8(+) (rho = 0.83, P <.0001) T cells. The M8 concentrations correlated weakly with both inhibition and nelfinavir concentrations. On the basis of our findings in lymphocytes from a healthy volunteer exposed to plasma from human immunodeficiency virus-positive patients, we believe it is likely that CD4(+) and CD8(+) lymphocytes in patients receiving nelfinavir as therapy for human immunodeficiency virus may have P-glycoprotein inhibited by plasma concentrations of nelfinavir. PMID- 12545147 TI - Teriparatide has no effect on the calcium-mediated pharmacodynamics of digoxin. AB - BACKGROUND: Teriparatide (recombinant human parathyroid hormone [1-34]) stimulates bone formation and causes small transient increases in serum calcium concentration. We assessed whether teriparatide causes a change in digoxin pharmacodynamic effects by measuring systolic time intervals and heart rate. METHODS: Measurements were made by echocardiographic Doppler that examined 3 systolic time intervals, as follows: QS(2) (time from Q wave on electrocardiogram to the closure of the aortic valve), left ventricular ejection time, and pre ejection period, all corrected for changes in heart rate. Fifteen healthy subjects (2 men and 13 women) were administered a single subcutaneous teriparatide dose (20 microg) on day 1 and then equilibrated on a daily oral dose of digoxin for 15 days. Subcutaneous placebo and teriparatide, 20 microg, were given in a randomized crossover design with the 14th (day 15) and 15th (day 16) digoxin doses. Serial systolic time interval and heart rate measurements were obtained on days 1, 15, and 16. RESULTS: After subjects were dosed to steady state with digoxin, there were statistically significant reductions in QS(2) corrected for heart rate (QS(2)c) of 23 to 25 ms and heart rate of 4 to 6 beats/min. However, there was no difference between treatment with digoxin plus placebo versus digoxin plus teriparatide. The study was powered to find a difference in QS(2)c as small as 6 ms (alpha =.05, beta =.2). CONCLUSION: Teriparatide, 20 microg subcutaneously, does not alter the cardiac effect of digoxin. PMID- 12545148 TI - Population pharmacodynamic analysis of octreotide in acromegalic patients. AB - INTRODUCTION: Octreotide is an octapeptide analog of somatostatin used to normalize growth hormone levels in acromegaly. This article presents a population analysis of the relationship between octreotide and growth hormone concentrations in 94 patients with acromegaly, including 10 patients responding incompletely to subcutaneous treatment (poor responders). METHODS: Growth hormone and octreotide concentrations were recorded hourly over 12-hour time periods during long-term subcutaneous treatment. Twelve-hour profiles were also collected on different days up to 2 months after intramuscular injection of the long-acting formulation. We modeled the inhibition of growth hormone secretion by octreotide with a direct maximum inhibition model. A joint analysis of both formulations was performed with NONMEM (GloboMax, LLC, Hanover, Md). During model building, we examined the relationships between parameters and demographic covariates or formulations with the use of likelihood ratio tests. RESULTS: The baseline growth hormone level was higher in poor responders and was best described by a bimodal distribution. The maximum inhibition was common to both formulations and had a mean of 90%, with low interindividual variability. Sensitivity to octreotide (50% inhibitory concentration) was found to be slightly lower on average with intramuscular administration than with subcutaneous administration. CONCLUSION: Given adequate doses of octreotide, in 72% of 94 patients, growth hormone would decrease to levels below 2.5 ng. mL(-1), considered to be a desirable target concentration in acromegaly. This study provides a way to identify poor responders during subcutaneous treatment, allowing an early clinical decision to be made to switch nonresponders to alternative therapies. PMID- 12545149 TI - Analgesic effects of morphine and morphine-6-glucuronide in a transcutaneous electrical pain model in healthy volunteers. AB - OBJECTIVE: Our objective was to quantify the extent and time course of the effects of morphine-6-glucuronide and morphine on pain threshold, pain tolerance, pupil diameter, and side effects. METHODS: In a double-blind, placebo-controlled, randomized, 3-way crossover study, 12 healthy volunteers (6 men and 6 women) received 63 to 112 mg of morphine-6-glucuronide or 26 to 66 mg of morphine as an intravenous bolus, followed by an infusion of the same medication for 1.8 to 6.4 hours. Analgesia was assessed every 30 minutes for up to 16 hours by means of transcutaneous electrical stimulation (sine wave, 5 Hz; intensity, 0-9.99 mA). Pupil diameter and side effects were recorded concomitantly. RESULTS: At the administered doses, morphine-6-glucuronide and morphine had comparable effects on pain tolerance, pupil diameter, and side effects. The delay between the time course of the plasma concentrations and the time course of the effects was longer for morphine-6-glucuronide than for morphine (transfer half-life, 8.2 hours versus 2.6 hours for pain tolerance and 7.7 hours versus 2.8 hours for pupil diameter). The slope of the linear concentration versus effect relationship for pain tolerance was flatter for morphine-6-glucuronide than for morphine (0.05% versus 0.6% increase in pain tolerance per nanomole per liter of morphine-6 glucuronide and morphine at effect site, respectively). Morphine-6-glucuronide was less potent than morphine in producing pupil constriction (mean concentration at half-maximum effect, 745 nmol/L versus 26.4 nmol/L for morphine-6-glucuronide and morphine, respectively). In carriers of the mutated G118 allele of the mu opioid receptor, the potency of the pupil-constricting effects of morphine-6 glucuronide and morphine was significantly smaller, and carriers of the G118 allele reported less nausea and vomited less often after administration of morphine-6-glucuronide. CONCLUSIONS: Morphine-6-glucuronide clearly produced analgesic effects in healthy volunteers. However, the high amounts of systemic morphine-6-glucuronide needed to produce the same effects as morphine suggest that morphine-6-glucuronide barely contributes to the central nervous opioid effects after administration of analgesic doses of morphine. PMID- 12545151 TI - Routine early therapeutic drug monitoring is necessary during the first week of life. PMID- 12545150 TI - Genetic variation in cyclooxygenase 1: effects on response to aspirin. AB - BACKGROUND: A critical clinical application of the Human Genome Project is to identify functional variation in genes related to disease or responses to xenobiotics. This study moved toward that goal by combining polymorphism detection with functional assays for the therapeutic target gene cyclooxygenase 1 (COX-1). Cyclooxygenase 1 (prostaglandin endoperoxide G/H synthase [PTGS1]) catalyzes the metabolism of arachidonic acid to prostaglandin H(2), which is subsequently metabolized to thromboxane A(2). METHODS: Thirty-eight healthy participants were enrolled in this study to correlate functional and genetic variation of cyclooxygenase 1. Arachidonic acid, with and without aspirin (acetylsalicylic acid) and ethanol pretreatment, was used to stimulate the formation of prostaglandin H(2), measured as prostaglandin F(2 alpha) (PGF(2 alpha)), in human platelets. The cyclooxygenase 1 complementary deoxyribonucleic acid coding sequence and genomic deoxyribonucleic acid upstream from the cyclooxygenase 1 transcription start site (2.9 kilobases) were sequenced in 38 individuals, with 9 single-nucleotide polymorphisms identified. RESULTS: Two single-nucleotide polymorphisms, A-842G and C50T, were in complete linkage disequilibrium. Participants who were heterozygous for the A-842G/C50T haplotype showed significantly (P =.01) greater inhibition of prostaglandin H(2) formation by acetylsalicylic acid (30 micromol/L) compared with common allele homozygotes. CONCLUSIONS: The discovery of a functional single-nucleotide polymorphism in the cyclooxygenase 1 locus may ultimately improve the safe and effective use of acetylsalicylic acid by better tailoring of dosage with an individual's genetic variation. PMID- 12545152 TI - Medical practice, race, and drug use. PMID- 12545153 TI - Stat3 activation regulates the expression of vascular endothelial growth factor and human pancreatic cancer angiogenesis and metastasis. AB - Expression of vascular endothelial growth factor (VEGF), a key angiogenic protein, has been linked with pancreatic cancer progression. However, the molecular basis for VEGF overexpression remains unclear. Immunohistochemical studies have indicated that VEGF overexpression coincides with elevated Stat3 activation in human pancreatic cancer specimens. In our study, more than 80% of the human pancreatic cancer cell lines used exhibited constitutively activated Stat3, with Stat3 activation correlated with the VEGF expression level. Blockade of activated Stat3 via ectopic expression of dominant-negative Stat3 significantly suppressed VEGF expression, angiogenesis, tumor growth, and metastasis in vivo. Furthermore, constitutively activated Stat3 directly activated the VEGF promoter, whereas dominant-negative Stat3 inhibited the VEGF promoter. A putative Stat3-responsive element on the VEGF promoter was identified using a protein-DNA binding assay and confirmed using a promoter mutagenesis assay. These results indicate that Stat3 directly regulates VEGF expression and hence angiogenesis, growth, and metastasis of human pancreatic cancer, suggesting that Stat3 signaling may be targeted for treatment of pancreatic cancer. PMID- 12545154 TI - RhoG regulates gene expression and the actin cytoskeleton in lymphocytes. AB - RhoG, a member of the Rho family of GTPases, has been implicated as a regulator of the actin cytoskeleton. In this study, we show a novel function for the small GTPase RhoG on the regulation of the interferon-gamma promoter and nuclear factor of activated T cells (NFAT) gene transcription in lymphocytes. Optimal function of RhoG for the expression of these genes requires a calcium signal, normally provided by the antigen receptor. In addition, RhoG potentiation of NFAT requires the indirect activity of Rac and Cdc42; however, pathways distinct from those activated by Rac and Cdc42 mediate RhoG activation of NFAT-dependent transcription. Using effector domain mutants of RhoG we found that its ability to potentiate NFAT-dependent transcription correlates with its capacity to increase actin polymerization, supporting the suggestion that NFAT-dependent transcription is an actin-dependent process. RhoG also promotes T-cell spreading on fibronectin, a property that is independent of its ability to enhance NFAT dependent transcription. Hence, these results implicate RhoG in leukocyte trafficking and the control of gene expression induced in response to antigen encounter. PMID- 12545155 TI - A novel PHD-finger motif protein, p47ING3, modulates p53-mediated transcription, cell cycle control, and apoptosis. AB - A candidate tumor suppressor gene, p33ING1, was previously identified by using the genetic suppressor element methodology. p33ING1 cooperates with p53 and plays a significant role in p53-mediated cellular processes. Recently, we have identified p33ING2, which shows a sequence homology similar to p33ING1 and modulates p53 function. In the present study, we identified and characterized another 'ING family' gene. The estimated molecular weight of the encoded protein is 46.8 kDa, thus, we named it p47ING3. The p47ING3 gene is located at chromosome 7q31.3 and consists of 12 exons that encode 418 amino acids. A computational domain search revealed a C-terminal PHD-finger motif. Such motifs are common in proteins involved in chromatin remodeling. p47ING3 is highly expressed in some normal human tissues or organs, including the spleen, testis, skeletal muscle, and heart. p47ING3 expression levels varied among cancer cell lines. p47ING3 overexpression resulted in a decreased population of cells in S phase, a diminished colony-forming efficiency, and induced apoptosis in RKO cells, but not in RKO-E6 cells with inactivated p53. p47ING3 activates p53-transactivated promoters, including promoters of p21/waf1 and bax. Thus, we have isolated a novel ING family gene, p47ING3, which modulates p53-mediated transcription, cell cycle control, and apoptosis. PMID- 12545156 TI - Myc represses differentiation-induced p21CIP1 expression via Miz-1-dependent interaction with the p21 core promoter. AB - Inhibition of cellular differentiation is one of the well-known biological activities of c-Myc-family proteins. We show here that Myc represses differentiation-induced expression of the cyclin-dependent kinase (CDK) inhibitor p21CIP1 (CDKN1A, p21), known to play an important role in cell fate decisions during growth and differentiation, in hematopoietic cells. Our results demonstrate that the c-Myc-responsive region is situated in the p21 core promoter. c-Myc binds to this region in vitro and in vivo through interaction with the initiator-binding Zn-finger transcription factor Miz-1, which associates directly with the promoter. Association of Myc with the promoter in vivo correlates inversely with p21 expression. Using mutants of c-Myc with impaired binding to Miz-1, our results further show that repression of p21 promoter/reporters as well as the endogenous p21 gene by Myc depends on interaction with Miz-1. Expression of Miz-1 increases during hematopoietic differentiation and Miz-1 activates the p21 promoter under conditions of low Myc levels, indicating a positive role for free Miz-1 in this process. In conclusion, repression of differentiation-induced p21 expression through Miz-1 may be an important mechanism by which Myc blocks differentiation. PMID- 12545157 TI - p27 deficiency desensitizes Rb-/- cells to signals that trigger apoptosis during pituitary tumor development. AB - Low p27 expression in many human cancers is a prognostic indicator for poor outcome. While analysing the mechanism by which p27 deficiency contributed to tumor development in the Rb+/- mouse model, we identified a role for p27 as a proapoptotic tumor suppressor. We examined the cell cycle and apoptotic response of these pituitary tumor cells to the dopamine analog bromocriptine as well as the expression of Arf and other cell cycle and apoptotic regulators in these tumors. We also examined the expression of Arf and its function in mouse embryo fibroblasts either singly or doubly deficient for Rb and p27. From these studies, we concluded that the absence of p27 disabled the trigger for an Arf-dependent apoptotic response in Rb-/- tumor cells. This suggests a novel mechanism by which the loss of p27 may impact on tumor development. PMID- 12545158 TI - Selective ablation of human cancer cells by telomerase-specific adenoviral suicide gene therapy vectors expressing bacterial nitroreductase. AB - Reactivation of telomerase maintains telomere function and is considered critical to immortalization in most human cancer cells. Elevation of telomerase expression in cancer cells is highly specific: transcription of both RNA (hTR) and protein (hTERT) components is strongly upregulated in cancer cells relative to normal cells. Therefore, telomerase promoters may be useful in cancer gene therapy by selectively expressing suicide genes in cancer cells and not normal cells. One example of suicide gene therapy is the bacterial nitroreductase (NTR) gene, which bioactivates the prodrug CB1954 into an active cytotoxic alkylating agent. We describe construction of adenovirus vectors harbouring the bacterial NTR gene under control of the hTR or hTERT promoters. Western blot analysis of NTR expression in normal and cancer cells infected with adenoviral vectors showed cancer cell-specific nitroreductase expression. Infection with adenoviral telomerase-NTR constructs in a panel of seven cancer cell lines resulted in up to 18-fold sensitization to the prodrug CB1954, an effect that was retained in two drug-resistant ovarian lines. Importantly, no sensitization was observed with either promoter in any of the four normal cell strains. Finally, an efficacious effect was observed in cervical and ovarian xenograft models following single intratumoural injection with low doses of vector, followed by injection with CB1954. PMID- 12545159 TI - Interferon regulatory factor-1 (IRF-1) is a mediator for interferon-gamma induced attenuation of telomerase activity and human telomerase reverse transcriptase (hTERT) expression. AB - Constitutive activation of the telomerase is a key step in the development of human cancers. Interferon-gamma (IFN-gamma) signaling induces growth arrest in many tumors through multiple regulatory mechanisms. In this study, we show that IFN-gamma signaling represses telomerase activity and human telomerase reverse transcriptase (hTERT) transcription, and suggest that this signaling is mediated by IRF-1. Ectopic expression of IRF-1 attenuated hTERT promoter activity. Murine embryonic fibroblasts (MEFs) genetically deficient in IRF-1 (IRF-1(-/-)) showed an elevated level (>15 times) of hTERT promoter activity as compared to the hTERT promoter activity of wild-type MEFs. The telomerase activity and hTERT expression in IRF-1(-/-) MEFs were downregulated by IRF-1 transfection. Interestingly, less extent of telomerase repression was observed in HPV E6 and E7 negative, p53 mutant HT-3 cells than in HPV 18 E6 and E7 positive HeLa cells (intact p53). These findings provide evidence that IRF-1 is a potential mediator of IFN-gamma induced attenuation of telomerase activity and hTERT expression. PMID- 12545160 TI - Downregulation of uPA inhibits migration and PI3k/Akt signaling in glioblastoma cells. AB - The ability of glioma cells to migrate great distances from a primary tumor mass is the primary cause of tumor recurrence. The urokinase-type plasminogen activator (uPA) is a serine protease that can initiate proteolytic cascades, which result in remodeling of extracellular matrix and basement membrane, allowing cells to move across and through these barriers. The binding between uPA and its receptor uPAR also mediates several signaling events that seem to contribute to the evolution of a migratory phenotype. In this study, we determined how the downregulation of uPA affects the signaling pathways leading to cell migration. Stably transfecting human glioblastoma cells with antisense uPA decreased the amount of cell-bound uPA and disrupted actin cytoskeleton formation and cell migration. The phosphatidylinositol 3-kinase (PI3k) and Akt signaling pathway has been suggested to mediate migration in various cancer cells. The antisense-uPA clones also had less phosphorylated PI3k and Akt than control cells, a finding associated with decreased cell migration, G2/M-phase arrest, and decreased clonogenic survival. Decreased activation of PI3k and the antiapoptotic factor Akt was not sufficient to induce apoptosis in the antisense uPA clones, but staurosporine sensitized them to apoptosis to a greater extent than control cells. These results indicate that PI3k/Akt pathway is involved in the signaling cascade required to induce cell migration and that uPA has a direct role in regulating migration. PMID- 12545161 TI - Ataxin-2 promotes apoptosis of human neuroblastoma cells. AB - Neuroblastoma is a highly heterogeneous tumor of young children. Although many advances have been made towards understanding the molecular mechanisms dictating the phenotypic heterogeneity, the prognosis of children with neuroblastoma, particularly of progressively growing variants, has remained dire. About 10% of neuroblastomas regress spontaneously, probably by apoptosis, while another 20% have amplified the MYCN gene resulting in a poor prognosis. In pursuit of identifying cell death-associated genes in neuroblastoma, we encountered the SCA2 gene, coding for ataxin-2, as an important player. Here, we report that enforced expression of wild-type ataxin-2, but not of mutant ataxin-2, sensitizes neuroblastoma cells for apoptosis. In line with this, higher levels of ataxin-2 were detected in apoptotic cells compared to nonapoptotic cells. Neuroblastoma tumors with amplified MYCN contain significantly less ataxin-2 protein than tumors without amplified MYCN. Collectively, our data suggest that ataxin-2 has an important role in regulating the susceptibility of neuroblastoma cells to apoptotic stimuli in vitro and in vivo. PMID- 12545162 TI - Transient activation of NF-kappaB through a TAK1/IKK kinase pathway by TGF-beta1 inhibits AP-1/SMAD signaling and apoptosis: implications in liver tumor formation. AB - NF-kappaB has been implicated in the regulation of apoptosis, a key mechanism of normal and malignant growth control. Previously, we demonstrated that inhibition of NF-kappaB activity by TGF-beta1 leads directly to induction of apoptosis of murine B-cell lymphomas and hepatocytes. Thus, we were surprised to determine that NF-kappaB is transiently activated in response to TGF-beta1 treatment. Here we elucidate the mechanism of TGF-beta1-mediated regulation of NF-kappaB and induction of apoptosis in epithelial cells. We report that TGF-beta1 activates IKK kinase, which mediates IkappaB-alpha phosphorylation. In turn, the activation of IKK following TGF-beta1 treatment is mediated by the TAK1 kinase. As a result of NF-kappaB activation, IkappaB-alpha mRNA and protein levels are increased leading to postrepression of NF-kappaB and induction of cell death. Inhibition of NF-kappaB following TGF-beta1 treatment increased AP-1 complex transcriptional activity through sustained c-Jun phosphorylation, thereby potentiating AP-1/SMADs mediated cell killing. Furthermore, TGF-beta1-mediated upregulation of Smad7 appeared independent of NF-kappaB. In hepatocellular carcinomas of TGF-beta1 or TGF-alpha/c-myc transgenic mice, we observed constitutive activation of NF-kappaB that led to inhibition of JNK signaling. Overall, our data illustrate an autocrine mechanism based on the ability of IKK/NF-kappaB/IkappaB-alpha signaling to negatively regulate NF-kappaB levels thereby permitting TGF-beta1-induced apoptosis through AP-1 activity. PMID- 12545163 TI - Pulmonary adenoma susceptibility 1 (Pas1) locus affects inflammatory response. AB - Two outbred mouse lines, phenotypically selected for differential subcutaneous (s.c.) acute inflammatory response (AIR), were analysed for urethane-induced lung inflammatory response and susceptibility to lung tumorigenesis. AIR(min) mice, which show a low response to s.c. acute inflammation, developed a persistent subacute lung inflammatory response and a 40-fold higher lung tumor multiplicity than did AIR(max) mice, which are selected for high response to s.c. acute inflammation and showed a transient lung inflammatory response. A highly significant linkage disequilibrium pattern was observed in AIR(max) and AIR(min) mice at marker alleles located within a 452-kb pulmonary adenoma susceptibility 1 (Pas1) locus region, thus defining the location of gene candidacy for inflammatory response and for the biological effects of Pas1 in this region. AIR(min) and AIR(max) mice segregated by descent the Pas1(s) and Pas1(r) alleles, respectively, providing evidence for the involvement of the Pas1 locus in the inflammatory response. The 452-kb region contains Kras2 and four additional genes, including the lymphoid-restricted membrane protein (Lrmp) gene, whose Pro- >Leu nonconservative variation was linked with inflammatory response and Pas1 allelotype. These results provide a model to explore the mechanism underlying inherited predisposition to lung cancer in the context of a link to inflammation. PMID- 12545164 TI - Bypass of telomere-dependent replicative senescence (M1) upon overexpression of Cdk4 in normal human epithelial cells. AB - Many stimuli causing 'stress' or DNA damage in cells can produce phenotypes that overlap with telomere-based replicative senescence. In epithelial systems, the p16/RB pathway may function as a stress senescence-signaling pathway independent of telomere shortening. Overexpressing cyclin-dependent kinase 4 (Cdk4) in human epidermal keratinocytes and human mammary epithelial cells not only prevents the p16(INK4a)-associated premature growth arrest due to telomere-independent stress (e.g., inadequate culture conditions), but also bypasses the ensuing telomere dependent senescence (M1). Overexpressed Cdk4 in epithelial cells induces a dramatic upregulation of p16(INK4a) and milder upregulation of p53 and p21(WAF1), which become unresponsive to UV irradiation. Despite the high levels of these checkpoint factors, Cdk4-overexpressing cells divide in an apparently normal regulated fashion, are able to respond to changes in calcium levels, retain the stem cell phenotype, and fully differentiate and stratify. These results suggest that the differentiation pathways in Cdk4-overexpressing cells remain intact. PMID- 12545165 TI - DLC-1 gene inhibits human breast cancer cell growth and in vivo tumorigenicity. AB - The human DLC-1 (deleted in liver cancer 1) gene was cloned from a primary human hepatocellular carcinoma (HCC) and mapped to the chromosome 8p21-22 region frequently deleted in common human cancers and suspected to harbor tumor suppressor genes. DLC-1 was found to be deleted or downregulated in a significant number of HCCs. We expanded our investigations to other cancers with recurrent deletions of 8p22, and in this study examined alterations of DLC-1 in primary human breast tumors, human breast, colon, and prostate tumor cell lines. Genomic deletion of DLC-1 was observed in 40% of primary breast tumors, whereas reduced or undetectable levels of DLC-1 mRNA were seen in 70% of breast, 70% of colon, and 50% of prostate tumor cell lines To see whether DLC-1 expression affects cell growth and tumorigenicity, two breast carcinoma cell lines lacking the expression of endogenous gene were transfected with the DLC-1 cDNA. In both cell lines, DLC 1 transfection caused significant growth inhibition and reduction of colony formation. Furthermore, introduction of the DLC-1 cDNA abolished the in vivo tumorigenicity in nude mice, suggesting that the DLC-1 gene plays a role in breast cancer by acting as a bona fide tumor suppressor gene. PMID- 12545166 TI - Role of DNA methylation in the suppression of Apaf-1 protein in human leukaemia. AB - Apaf-1 protein deficiency occurs in human leukaemic blasts and confers resistance to cytochrome-c-dependent apoptosis. Demethylation treatment with 5-aza-2' deoxycytidine (5aza2dc) increased the sensitivity of the K562 leukaemic cell line to UV light-induced apoptosis in association with increased Apaf-1 protein levels. There was no correlation between Apaf-1 protein expression and Apaf-1 mRNA levels after the demethylation treatment. Methylation-specific polymerase chain reaction was used to show that the methylation can occur within the Apaf-1 promoter region in leukaemic blasts. Apaf-1 DNA methylation was demonstrated in acute myeloid leukaemia, chronic myeloid leukaemia and acute lymphoid leukaemia, suggesting that it is not specific to a particular leukaemia subtype. Apaf-1 protein expression did not correlate with Apaf-1 mRNA levels in human leukaemic blasts. Some leukaemic cells expressed high levels of Apaf-1 mRNA but low levels of Apaf-1 protein. This study suggests that Apaf-1 DNA promoter methylation might contribute to the inactivation of Apaf-1 expression. However, Apaf-1 protein levels might also be controlled at post-transcription level. PMID- 12545167 TI - ID2 expression in neuroblastoma does not correlate to MYCN levels and lacks prognostic value. AB - The MYCN proto-oncogene is frequently amplified in a subgroup of highly aggressive neuroblastomas. The molecular mechanism(s) by which the overexpressed MYCN contributes to an aggressive tumor cell behavior is not well understood. Recently, it was reported that the ID2 gene is a direct target for the MYCN and MYC transcription factors, and that ID2 expression and MYCN amplification correlate positively in neuroblastoma. In addition, ID2 expression was proposed as a negative prognostic parameter. As these results are of potential clinical importance, but not in agreement with our own initial observations, the putative correlation between ID2 and MYC(N) expression in neuroblastoma cell lines and tumors was reinvestigated. We found no correlation between MYCN and ID2 expression in neuroblastoma cell lines or tumor specimens. However, we did find a significant positive correlation between MYC and ID2 expressions in both MYCN amplified and single-copy tumor specimens, and in MYCN single-copy cell lines. As previously reported, we also found an inverse correlation between MYC and MYCN expressions. Importantly, we could not confirm the reported prognostic power of ID2-expression in neuroblastoma. These data, obtained in two independent laboratories, challenge the previously proposed ID2-MYCN relation. PMID- 12545168 TI - Frequent epigenetic inactivation of the RASSF1A tumour suppressor gene in testicular tumours and distinct methylation profiles of seminoma and nonseminoma testicular germ cell tumours. AB - Testicular germ cell tumours (TGCTs) are histologically heterogeneous neoplasms with variable malignant potential. Previously, we demonstrated frequent 3p allele loss in TGCTs, and recently we and others have shown that the 3p21.3 RASSF1A tumour suppressor gene (TSG) is frequently inactivated by promoter hypermethylation in a wide range of cancers including lung, breast, kidney and neuroblastoma. In order to investigate the role of epigenetic events in the pathogenesis of TGCTs, we analysed the promoter methylation status of RASSF1A and nine other genes that may be epigenetically inactivated in cancer (p16(INK4A), APC, MGMT, GSTP1, DAPK, CDH1, CDH13, RARbeta and FHIT) in 24 primary TGCTs (28 histologically distinct components). RASSF1A methylation was detected in four of 10 (40%) seminomas and 15 of 18 (83%) nonseminoma TGCT (NSTGCT) components (P=0.0346). None of the other nine candidate genes were methylated in seminomas, but MGMT (44%), APC (29%) and FHIT (29%) were frequently methylated in NSTGCTs. Furthermore, in two mixed germ cell tumours, the NSTGCT component for one demonstrated RASSF1A, APC and CDH13 promoter methylation, but the seminoma component was unmethylated for all genes analysed. In the second mixed germ cell tumour, the NSTGCT component was methylated for RASSF1A and MGMT, while the seminoma component was methylated only for RASSF1A. In all, 61% NSTGCT components but no seminoma samples demonstrated promoter methylation at two or more genes (P=0.0016). These findings are consistent with a multistep model for TGCT pathogenesis in which RASSF1A methylation occurs early in tumorigenesis and additional epigenetic events characterize progression from seminoma to NSTGCTs. PMID- 12545169 TI - An LOH and mutational investigation of the ST7 gene locus in human esophageal carcinoma. AB - Frequent loss of heterozygosity (LOH) on human chromosome 7q31 has been reported in numerous malignancies. Suppressor of tumorigenicity 7 (ST7) has been identified as a candidate tumor suppressor gene in this region. To identify whether 7q31 and genetic alterations of ST7 were involved in human esophageal carcinogenesis, we performed LOH mapping of a 5.4 cM region at 7q31-q35 in 43 primary esophageal carcinomas, as well as mutational analyses of the ST7 gene in tumors with LOH in this region. Of 43 tumors, 12 (28%) showed LOH at 7q31-q35. These included four (22%) of 18 squamous cell carcinomas and eight (32%) of 25 adenocarcinomas. The peak LOH locus was D7S480, lying 4.2 Mb telomeric to ST7 and showing LOH in eight of 37 informative tumors, or 22%. No mutations were found in the entire coding or flanking intronic regions of the ST7 gene among 12 tumors with 7q-LOH. In addition, quantitative RT-PCR analyses of ST7 mRNA expression levels in 11/13 normal-tumor pairs failed to show more than a 50% decrease in tumor ST7 mRNA relative to matched normal tissues. These data suggest that LOH at 7q31-q35 is involved in the origin or progression of at least a subset of esophageal carcinomas, but that ST7 is not the target gene of this somatic event. PMID- 12545170 TI - Aberrant sensing of extracellular Ca2+ by cultured ataxia telangiectasia fibroblasts. AB - Ataxia telangiectasia (AT) is a human hereditary syndrome whose underlying gene product, ataxia telangiectasia mutated (ATM) protein kinase, is involved in multiple intracellular signaling pathways. We demonstrated previously that AT fibroblasts are defective in intracellular Ca(2+) mobilization in response to both stress-inducing and mitogenic stimuli. To extend these findings, normal and AT cells were exposed to serum in the presence of different concentrations of extracellular Ca(2+) ([Ca(2+)](o)), and release of intracellular Ca(2+), activation of calmodulin-dependent protein kinase II and phosphorylation of kinases ERK1 and 2 were monitored. When maintained in high [Ca(2+)](o) (0.42 mM), normal fibroblasts responded to serum introduction more rapidly and efficiently than did AT cells. Unexpectedly, decreasing the [Ca(2+)](o) in the medium had a diametrically opposite effect. Under low [Ca(2+)](o) (0.0022 mM) conditions, normal cells were slow and inefficient in their responses, whereas AT cells showed a substantial improvement in all three end points. These findings demonstrate that loss of ATM kinase function deregulates the extracellular calcium-sensing receptor (CaR). This malfunction presumably arises from a post transcriptional event, since CaR mRNA proved to be normal in AT cells. Together, our data suggest that ATM may mediate cell response to mitogenic factors by tightly regulating the set point of the CaR and thereby modulating the crosstalk between this metabotropic receptor and growth factor receptors. Alternatively, the faulty sensing of extracellular calcium in AT cells may be secondary to a state of chronic oxidative stress attributable to ATM deficiency. PMID- 12545171 TI - Fas engagement induces neurite growth through ERK activation and p35 upregulation. AB - Fas (also known as CD95), a member of the tumour-necrosis receptor factor family of 'death receptors', can induce apoptosis or, conversely, can deliver growth stimulatory signals. Here we report that crosslinking Fas on primary sensory neurons induces neurite growth through sustained activation of the extracellular signal regulated kinase (ERK) pathway and the consequent upregulation of p35, a mediator of neurite outgrowth. In addition, functional recovery after sciatic nerve injury is delayed in Fas-deficient lpr mice and accelerated by local administration of antibodies against Fas, which indicates that Fas engagement may contribute to nerve regeneration in vivo. Our findings define a role for Fas as an inducer of both neurite growth in vitro and accelerated recovery after nerve injury in vivo. PMID- 12545172 TI - Three-dimensional analysis of post-Golgi carrier exocytosis in epithelial cells. AB - Targeted delivery of proteins to distinct plasma membrane domains is critical to the development and maintenance of polarity in epithelial cells. We used confocal and time-lapse total internal reflection fluorescence microscopy (TIR-FM) to study changes in localization and exocytic sites of post-Golgi transport intermediates (PGTIs) carrying GFP-tagged apical or basolateral membrane proteins during epithelial polarization. In non-polarized Madin Darby Canine Kidney (MDCK) cells, apical and basolateral PGTIs were present throughout the cytoplasm and were observed to fuse with the basal domain of the plasma membrane. During polarization, apical and basolateral PGTIs were restricted to different regions of the cytoplasm and their fusion with the basal membrane was completely abrogated. Quantitative analysis suggested that basolateral, but not apical, PGTIs fused with the lateral membrane in polarized cells, correlating with the restricted localization of Syntaxins 4 and 3 to lateral and apical membrane domains, respectively. Microtubule disruption induced Syntaxin 3 depolarization and fusion of apical PGTIs with the basal membrane, but affected neither the lateral localization of Syntaxin 4 or Sec6, nor promoted fusion of basolateral PGTIs with the basal membrane. PMID- 12545173 TI - Binding of SAP SH2 domain to FynT SH3 domain reveals a novel mechanism of receptor signalling in immune regulation. AB - SAP (or SH2D1A), an adaptor-like molecule expressed in immune cells, is composed almost exclusively of a Src homology 2 (SH2) domain. In humans, SAP is mutated and either absent or non-functional in X-linked lymphoproliferative (XLP) syndrome, a disease characterized by an inappropriate response to Epstein-Barr virus (EBV) infection. Through its SH2 domain, SAP associates with tyrosines in the cytoplasmic domain of the SLAM family of immune cell receptors, and is absolutely required for the function of these receptors. This property results from the ability of SAP to promote the selective recruitment and activation of FynT, a cytoplasmic Src-related protein tyrosine kinase (PTK). Here, we demonstrate that SAP operates in this pathway by binding to the SH3 domain of FynT, through a second region in the SAP SH2 domain distinct from the phosphotyrosine-binding motif. We demonstrate that this interaction is essential for SAP-mediated signalling in T cells, and for the capacity of SAP to modulate immune cell function. These observations characterize a biologically important signalling mechanism in which an adaptor molecule composed only of an SH2 domain links a receptor devoid of intrinsic catalytic activity to the kinase required for its function. PMID- 12545175 TI - Repeated phosphopeptide motifs in Claspin mediate the regulated binding of Chk1. AB - In vertebrates, the checkpoint-regulatory kinase Chk1 mediates cell-cycle arrest in response to a block in DNA replication or to DNA damaged by ultraviolet radiation. The activation of Chk1 depends on both Claspin and the upstream regulatory kinase ATR. Claspin is a large acidic protein that becomes phosphorylated and binds to Chk1 in the presence of checkpoint-inducing DNA templates in Xenopus egg extracts. Here we identify, by means of deletion analysis, a region of Claspin of 57 amino acids that is both necessary and sufficient for binding to Xenopus Chk1. This Chk1-binding domain contains two highly conserved repeats of approximately ten amino acids. A serine residue in each repeat (serine 864 and serine 895) undergoes phosphorylation during a checkpoint response. A mutant of Claspin containing non-phosphorylatable amino acids at positions 864 and 895 cannot bind to Chk1 and is unable to mediate its activation. Our results indicate that two phosphopeptide motifs in Claspin are essential for checkpoint signalling. PMID- 12545174 TI - SAP couples Fyn to SLAM immune receptors. AB - SAP (SLAM-associated protein) is a small lymphocyte-specific signalling molecule that is defective or absent in patients with X-linked lymphoproliferative syndrome (XLP). Consistent with its single src homology 2 (SH2) domain architecture and unusually high affinity for SLAM (also called CD150), SAP has been suggested to function by blocking binding of SHP-2 or other SH2-containing signalling proteins to SLAM receptors. Additionally, SAP has recently been shown to be required for recruitment and activation of the Src-family kinase FynT after SLAM ligation. This signalling 'adaptor' function has been difficult to conceptualize, because unlike typical SH2-adaptor proteins, SAP contains only a single SH2 domain and lacks other recognized protein interaction domains or motifs. Here, we show that the SAP SH2 domain binds to the SH3 domain of FynT and directly couples FynT to SLAM. The crystal structure of a ternary SLAM-SAP-Fyn SH3 complex reveals that SAP binds the FynT SH3 domain through a surface-surface interaction that does not involve canonical SH3 or SH2 binding interactions. The observed mode of binding to the Fyn-SH3 domain is expected to preclude the auto inhibited conformation of Fyn, thereby promoting activation of the kinase after recruitment. These findings broaden our understanding of the functional repertoire of SH3 and SH2 domains. PMID- 12545176 TI - Dlg, Scrib and Lgl regulate neuroblast cell size and mitotic spindle asymmetry. AB - Asymmetric cell division is important in generating cell diversity from bacteria to mammals. Drosophila melanogaster neuroblasts are a useful model system for investigating asymmetric cell division because they establish distinct apical basal cortical domains, have an asymmetric mitotic spindle aligned along the apical-basal axis, and divide unequally to produce a large apical neuroblast and a small basal daughter cell (GMC). Here we show that Discs large (Dlg), Scribble (Scrib) and Lethal giant larvae (Lgl) tumour suppressor proteins regulate multiple aspects of neuroblast asymmetric cell division. Dlg/Scrib/Lgl proteins show apical cortical enrichment at prophase/metaphase, and then have a uniform cortical distribution. Mutants have defects in basal protein targeting, a reduced apical cortical domain and reduced apical spindle size. Defects in apical cell and spindle pole size result in symmetric or inverted neuroblast cell divisions. Inverted divisions correlate with the appearance of abnormally small neuroblasts and large GMCs, showing that neuroblast/GMC identity is more tightly linked to cortical determinants than cell size. We conclude that Dlg/Scrib/Lgl are important in regulating cortical polarity, cell size asymmetry and mitotic spindle asymmetry in Drosophila neuroblasts. PMID- 12545177 TI - Direct interaction of two polarity complexes implicated in epithelial tight junction assembly. AB - Tight junctions help establish polarity in mammalian epithelia by forming a physical barrier that separates apical and basolateral membranes. Two evolutionarily conserved multi-protein complexes, Crumbs (Crb)-PALS1 (Stardust) PATJ (DiscsLost) and Cdc42-Par6-Par3-atypical protein kinase C (aPKC), have been implicated in the assembly of tight junctions and in polarization of Drosophila melanogaster epithelia. Here we identify a biochemical and functional link between these two complexes that is mediated by Par6 and PALS1 (proteins associated with Lin7). The interaction between Par6 and PALS1 is direct, requires the amino terminus of PALS1 and the PDZ domain of Par6, and is regulated by Cdc42 GTP. The transmembrane protein Crb can recruit wild-type Par6, but not Par6 with a mutated PDZ domain, to the cell surface. Expression of dominant-negative PALS1 associated tight junction protein (PATJ) in MDCK cells results in mis localization of PALS1, members of the Par3-Par6-aPKC complex and the tight junction marker, ZO-1. Similarly, overexpression of Par6 in MDCK cells inhibits localization of PALS1 to the tight junction. Our data highlight a previously unrecognized link between protein complexes that are essential for epithelial polarity and formation of tight junctions. PMID- 12545179 TI - HHS, USDA establish new regulations for use of select biological agents. PMID- 12545180 TI - Anesthetics in GEM: does TBE make the grade? PMID- 12545181 TI - Anemic virgin female rabbits. PMID- 12545183 TI - Housing and husbandry of Xenopus for oocyte production. AB - Despite their importance as a research model, particularly in developmental toxicology investigations, there are few established standards for maintaining Xenopus spp. frogs in the laboratory. The authors review the literature on handling, housing, nutrition, and breeding of Xenopus spp. for optimal oocyte production. PMID- 12545184 TI - Using site assessment and risk analysis to plan and build disaster-resistant programs and facilities. AB - Research animals need protection from the catastrophic losses that can occur during disasters. The best method of protecting animals is to house them in facilities that have been reinforced using disaster-resistant construction methods. The authors explain the use of site-specific risk analysis to determine the proper building site and features. PMID- 12545185 TI - Dual tail catheters for infusion and sampling in rats as an efficient platform for metabolic experiments. AB - The authors present a technique for establishing a virtually stress-free dual catheter system in rats, allowing researchers to conduct extended, complex experimental procedures involving the simultaneous infusion of multiple compounds and frequent collection of blood samples. PMID- 12545186 TI - Myosins and pathology: genetics and biology. AB - This article summarizes current knowledge on the genetics and possible molecular mechanisms of Human pathologies resulted from mutations within the genes encoding several myosin isoforms. Mutations within the genes encoding some myosin isoforms have been found to be responsible for blindness (myosins III and VIIA), deafness (myosins I, IIA, IIIA, VI, VIIA and XV) and familial hypertrophic cardiomyopathy (beta cardiac myosin heavy chain and both the regulatory and essential light chains). Myosin III localizes predominantly to photoreceptor cells and is proved to be engaged in the vision process in Drosophila. In the inner ear, myosin I is postulated to play a role as an adaptive motor in the tip links of stereocilia of hair cells, myosin IIA seems to be responsible for stabilizing the contacts between adjacent inner ear hair cells, myosin VI plays a role as an intracellular motor transporting membrane structures within the hair cells while myosin VIIA most probably participates in forming links between neighbouring stereocilia and myosin XV probably stabilizes the stereocilia structure. About 30% of patients with familial hypertrophic cardiomyopathy have mutations within the genes encoding the beta cardiac myosin heavy chain and both light chains that are grouped within the regions of myosin head crucial for its functions. The alterations lead to the destabilization of sarcomeres and to a decrease of the myosin ATPase activity and its ability to move actin filaments. PMID- 12545187 TI - Structural determinants of cooperativity in acto-myosin interactions. AB - Regulation of muscle contraction is a very cooperative process. The presence of tropomyosin on the thin filament is both necessary and sufficient for cooperativity to occur. Data recently obtained with various tropomyosin isoforms and mutants help us to understand better the structural requirements in the thin filament for cooperative protein interactions. Forming an end-to-end overlap between neighboring tropomyosin molecules is not necessary for the cooperativity of the thin filament activation. When direct contacts between tropomyosin molecules are disrupted, the conformational changes in the filament are most probably transmitted cooperatively through actin subunits, although the exact nature of these changes is not known. The function of tropomyosin ends, alternatively expressed in various isoforms, is to confer specific actin affinity. Tropomyosin's affinity or actin is directly related to the size of the apparent cooperative unit defined as the number of actin subunits turned into the active state by binding of one myosin head. Inner sequences of tropomyosin, particularly actin-binding periods 3 to 5, play crucial role in myosin-induced activation of the thin filament. A plausible mechanism of tropomyosin function in this process is that inner tropomyosin regions are either specifically recognized by myosin or they define the right actin conformation required for tropomyosin movement from its blocking position. PMID- 12545188 TI - Directionality of kinesin motors. AB - Kinesins are molecular motors that transport various cargoes along microtubule tracks using energy derived from ATP hydrolysis. Although the motor domains of kinesins are structurally similar, the family contains members that move on microtubules in opposite directions. Recent biochemical and biophysical studies of several kinesins make it possible to identify structural elements responsible for the different directionality, suggesting that reversal of the motor movement can be achieved through small, local changes in the protein structure. PMID- 12545189 TI - Actin in human colon adenocarcinoma cells with different metastatic potential. AB - Four human colon adenocarcinoma cell line variants with different metastatic potential were used to examine whether a correlation exists between actin level, state of actin polymerization and invasiveness of tumour cells. Monomeric (G), total (T) and filamentous (F) actin were determined in the cytosolic fraction of these cells. A statistically significant decrease in G actin level and increase in the state of actin polymerization (measured by F:G actin ratio) were found in the cytosol of three cell variants with higher metastatic potential and invasiveness (EB3, 3LNLN, 5W) compared with the parental cell line (LS180). Our experimental data lead to the conclusion that there is a correlation between the metastatic capacity of human colon adenocarcinoma cells and the state of actin polymerization. PMID- 12545190 TI - Myosin molecule packing within the vertebrate skeletal muscle thick filaments. A complete bipolar model. AB - Computer modelling related to the real dimensions of both the whole filament and the myosin molecule subfragments has revealed two alternative modes for myosin molecule packing which lead to the head disposition similar to that observed by EM on the surface of the cross-bridge zone of the relaxed vertebrate skeletal muscle thick filaments. One of the modes has been known for three decades and is usually incorporated into the so-called three-stranded model. The new mode differs from the former one in two aspects: (1) myosin heads are grouped into asymmetrical cross-bridge crowns instead of symmetrical ones; (2) not the whole myosin tail, but only a 43-nm C-terminus of each of them is straightened and near parallel to the filament axis, the rest of the tail is twisted. Concurrent exploration of these alternative modes has revealed their influence on the filament features. The parameter values for the filament models as well as for the building units depicting the myosin molecule subfragments are verified by experimental data found in the literature. On the basis of the new mode for myosin molecule packing a complete bipolar structure of the thick filament is created. PMID- 12545191 TI - The vertebrate skeletal muscle thick filaments are not three-stranded. Reinterpretation of some experimental data. AB - Computer simulation of mass distribution within the model and Fourier transforms of images depicting mass distribution are explored for verification of two alternative modes of the myosin molecule arrangement within the vertebrate skeletal muscle thick filaments. The model well depicting the complete bipolar structure of the thick filament and revealing a true threefold-rotational symmetry is a tube covered by two helices with a pitch of 2 x 43 nm due to arrangement of the myosin tails along a helical path and grouping of all myosin heads in the crowns rotated by 240 degrees and each containing three cross bridges separated by 0 degrees, 120 degrees, and 180 degrees. The cross-bridge crown parameters are verified by EM images as well as by optical and low-angle X ray diffraction patterns found in the literature. The myosin tail arrangement, at which the C-terminus of about 43-nm length is near-parallel to the filament axis and the rest of the tail is quite strongly twisted around, is verified by the high-angle X-ray diffraction patterns. A consequence of the new packing is a new way of movement of the myosin cross-bridges, namely, not by bending in the hinge domains, but by unwrapping from the thick filament surface towards the thin filaments along a helical path. PMID- 12545192 TI - Transport functions and physiological significance of 76 kDa Ral-binding GTPase activating protein (RLIP76). AB - We have recently demonstrated that a previously known Ral-binding GTPase activating protein, RLIP76, can also catalyze ATP-dependent transport of various structurally unrelated xeno- and endobiotics irrespective of their net charge (Awasthi et al., 2000, Biochemistry, 39: 9327). RLIP76 is a non-ATP binding cassette (ABC) protein but it has two ATP-binding sites and shows basal ATPase activity which is stimulated in the presence of its transport substrates (allocrites) such as doxorubicin (DOX) and S-(2,4-dinitrophenyl) glutathione (DNP SG). Proteoliposomes reconstituted with purified RLIP76 catalyze ATP-dependent, saturable transport of DOX, as well as of glutathione-conjugates including leukotrienes (LTC4) and the GSH-conjugate of 4-hydroxynonenal (GS-HNE). In erythrocytes the majority of transport activity for DOX, GS-HNE, and LTC4 is accounted for by RLIP76. Cells exposed to mild oxidative stress show a rapid and transient induction of RLIP76 resulting in an increased efflux of GS-HNE and acquire resistance to oxidative stress mediated toxicity and apoptosis. Cells transfected with RLIP76 acquire resistance to DOX through increased efflux of the drug suggesting its possible role in the mechanisms of drug-resistance. In this article, we discuss the significance of transport functions of RLIP76 highlighting its role in the defense mechanisms against oxidative injury, and modulation of signaling mechanisms. PMID- 12545193 TI - Transmembrane segment M2 of glycine receptor as a model system for the pore forming structure of ion channels. AB - The glycine receptor belongs to the ligand-gated ion channel superfamily. It is a chloride conducting channel composed of four transmembrane domains. It was previously shown that the second transmembrane domain (M2) of the glycine receptor forms an ion conduction pathway throughout lipid bilayers. The amino acid sequence of the transmembrane segment M2 of the glycine receptor has a high homology to all receptors of the ligand-gated ion channel superfamily. In our report, we have used a synthetic M2 peptide. It was incorporated into a planar membrane of known lipid composition and currents induced by M2 were measured by the Black Lipid Membrane technique. When the planar lipid bilayer was composed of 75% phosphatidylethanolamine and 25% phosphatidylserine, the reversal potential measured in a 150/600 mM KCl (cis/trans) gradient was -19 mV suggesting that the examined >pore was preferential to anions, P(K)/P(Cl) = 0.25. In contrast, when 75% phosphatidylserine and 25% phosphatidylethanolamine was used, the reversal potential was +20 mV and the >pore was preferential to cations, P(K)/P(Cl) = 4.36. Single-channel currents were recorded with two predominant amplitudes corresponding to the main-conductance and sub-conductance states. Both conductance states (about 12 pS and 30 pS) were measured in a symmetric solution of 50 mM KCl. The observed single-channel properties suggest that the selectivity and conductance of the pore formed by the M2 peptide of the glycine receptor depend on the lipid composition of the planar bilayer. PMID- 12545194 TI - Cross-talk between the ATP and ADP nucleotide receptor signalling pathways in glioma C6 cells. AB - In this review we summarize the present status of our knowledge on the enzymes involved in the extracellular metabolism of nucleotides and the receptors involved in nucleotide signalling. We focus on the mechanism of the ATP and ADP signalling pathways in glioma C6, representative of the type of nonexcitable cells. In these cells, ATP acts on the P2Y(2) receptor coupled to phospholipase C, whereas ADP on two distinct P2Y receptors: P2Y(1) and P2Y(12). The former is linked to phospholipase C and the latter is negatively coupled to adenylyl cyclase. The possible cross-talk between the ATP-, ADP- and adenosine-induced pathways, leading to simultaneous regulation of inositol 1,4,5-trisphosphate and cAMP mediated signalling, is discussed. PMID- 12545195 TI - Hydrolysis of cyclic GMP in rat peritoneal macrophages. AB - Intact rat peritoneal macrophages (rPM) treated with 3-isobutyl-1-methylxanthine (IBMX), an inhibitor of phosphodiesterases (PDEs), accumulated more cGMP than untreated cells. A PDE activity toward [(3)H]cGMP was detected in the soluble and particulate fractions of rPM. The hydrolysis of cGMP was Ca(2+)/calmodulin independent but increased in the presence of cGMP excess. Similar results were obtained when [(3)H]cAMP was used as a substrate. The hydrolytic activity towards both nucleotides was inhibited in the presence of IBMX. Therefore, the PDEs of families 2, 5, 10 and 11 are potential candidates for cGMP hydrolysis in the rPM. They may not only regulate the cGMP level in a feedback-controlled way but also link cGMP-dependent pathways with those regulated by cAMP. PMID- 12545196 TI - p19 detected in the rat retina and pineal gland is a guanylyl cyclase-activating protein (GCAP). AB - The Ca(2+)-dependent activation of retina-specific guanylyl cyclase (retGC) is mediated by guanylyl cyclase-activating proteins (GCAPs). Here we report for the first time detection of a 19 kDa protein (p19) with GCAP properties in extracts of rat retina and pineal gland. Both extracts stimulate synthesis of cGMP in rod outer segment (ROS) membranes at low (30 nM) but not at high (1 microM) concentrations of Ca(2+). At low Ca(2+), immunoaffinity purified p19 activates guanylyl cyclase(s) in bovine ROS and rat retinal membranes. Moreover, p19 is recognized by antibodies against bovine GCAP1 and, similarly to other GCAPs, exhibits a Ca(2+)-dependent electrophoretic mobility shift. PMID- 12545197 TI - Protective role of L-methionine against free radical damage of rat brain synaptosomes. AB - Incubation of rat brain synaptosomal/mitochondrial fraction with tert butylhydroperoxide resulted in accumulation of the lipid peroxidation product, conjugated dienes, damage of the synaptosomal membrane as evidenced by leakage of lactate dehydrogenase, and decrease of the total content of glutathione and of the GSH/GSSG ratio. This treatment also produced a considerable decrease of the ouabain-sensitive ATPase activity and a much smaller diminution of the activities of glutathione reductase and glutathione transferase. Preincubation of the synaptosomal/mitochondrial fraction with 0.5 or 1.0 mM L-methionine significantly protected against lipid peroxidation, membrane damage and changes in the glutathione system produced by low (1 mM) concentrations of tert butylhydroperoxide and completely prevented inactivation of ouabain-sensitive ATPase, glutathione reductase and glutathione transferase by such treatment. The importance of L-methionine in antioxidant protection is discussed. PMID- 12545198 TI - Effect of natural phenols on the catalytic activity of cytochrome P450 2E1. AB - The effect of protocatechuic acid, tannic acid and trans-resveratrol on the activity of p-nitrophenol hydroxylase (PNPH), an enzymatic marker of CYP2E1, was examined in liver microsomes from acetone induced mice. trans-Resveratrol was found to be the most potent inhibitor (IC(50) = 18.5 +/- 0.4 microM) of PNPH, while protocatechuic acid had no effect on the enzyme activity. Tannic acid with IC(50) = 29.6 +/- 3.3 microM showed mixed- and trans-resveratrol competitive inhibition kinetics (K(i) = 1 microM and 2.1 microM, respectively). Moreover, trans-resveratrol produced a NADPH-dependent loss of PNPH activity, suggesting mechanism-based CYP2E1 inactivation. These results indicate that trans resveratrol and tannic acid may modulate cytochrome P450 2E1 and influence the metabolic activation of xenobiotics mediated by this P450 isoform. PMID- 12545199 TI - Species- and substrate-specific stimulation of human plasma paraoxonase 1 (PON1) activity by high chloride concentration. AB - Paraoxonase 1 (PON1), contained in plasma high-density lipoproteins, plays an important role in the protection of plasma lipoproteins and cell membranes from oxidative damage. Previous studies indicate that human PON1 is stimulated by high NaCl concentrations. The aim of this study was to characterize in more detail the effect of salts on serum PON1. Paraoxon-hydrolyzing activity of human serum was stimulated by 81.6% following the addition of 1 M NaCl. The effect of NaCl was dose-dependent between 0.5 and 2 M. PON1 activity toward phenyl acetate was reduced by 1 M NaCl by 55.2%. Both the paraoxon- and phenyl acetate-hydrolysing activity was slightly lower in heparinized plasma than in serum, but NaCl had similar stimulatory and inhibitory effects on these activities, respectively. In rat, rabbit, and mouse, NaCl reduced PON1 activity. KCl had a similar effect on human PON1 as NaCl. Sodium nitrite also stimulated human PON1 but much less effectively than chloride salts. In contrast, sucrose, sodium acetate and sodium lactate had no significant effect. NaBr was a less effective PON1 activator than NaCl, whereas the effect of NaJ was non-significant. The activity of human PON1 toward homogentisic acid lactone and gamma-decanolactone was unaltered by NaCl. These data indicate that: 1) high concentrations of chlorides stimulate human PON1 activity toward paraoxon but not other substrates, 2) PON1 is inhibited by Cl(-) in other mammalian species, 3) the potency of human PON1 activation by halogene salts increases with decreasing atomic mass of the halide anion. PMID- 12545200 TI - Mouse cytosolic acetyl-CoA hydrolase, a novel candidate for a key enzyme involved in fat metabolism: cDNA cloning, sequencing and functional expression. AB - A cytosolic acetyl-CoA hydrolase (CACH) cDNA has been isolated from mouse liver cDNA library and sequenced. Recombinant expression of the cDNA in insect cells resulted in overproduction of active acetyl-CoA hydrolyzing enzyme protein. The mouse CACH cDNA encoded a 556-amino-acid sequence that was 93.5% identical to rat CACH, suggesting a conserved role for this enzyme in the mammalian liver. Database searching shows no homology to other known proteins, but reveals homological cDNA sequences showing two single-nucleotide polymorphisms (SNPs) in the CACH coding region. The discovery of mouse CACH cDNA is an important step towards genetic studies on the functional analysis of this enzyme by gene knockout and transgenic approaches. PMID- 12545201 TI - Protein kinases CKI and CKII are implicated in modification of ribosomal proteins of the yeast Trichosporon cutaneum. AB - Phosphorylation of acidic ribosomal proteins P1/P2-P0 is a common phenomenon in eukaryotic organisms. It was found previously that in Trichosporon cutaneum, unlike in other yeast species, in addition to the two acidic ribosomal proteins, two other proteins of 15 kDa and 19 kDa of the small ribosomal subunit were phosphorylated. Here we describe two protein kinases: CKI and CKII, which are engaged in the modification of T. cutaneum ribosomal proteins. The acidic ribosomal proteins and the protein of 19 kDa were modified by CKII associated with ribosomes, while the protein of 15 kDa was modified by CKI. Protein kinase CKI was purified from cell-free extract (CKIC) and from ribosomal fraction (CKIR). The molecular mass of CKIC was established at 33 kDa while that of CKIR at 35-37 kDa. A protein of 40 kDa copurified with CKIR but not CKIC. Heparin significantly increased 40 kDa protein phosphorylation level by CKIR. Microsequencing analysis revealed the presence of CKI recognition motifs in the N terminal fragment of the 40 kDa protein. PMID- 12545202 TI - The effect of vanadate on Pichia pastoris growth, protein kinase A activity and ribosomal protein phosphorylation. AB - It was found that wild type yeast Pichia pastoris can tolerate vanadate concentration as high as 25 mM in the growth medium. Moreover, four vanadate resistant P. pastoris strains designated JC100/1, JC100/3, JC100/9 and JC100/15 exhibiting tolerance up to 150 mM vanadate were selected. Growth of P. pastoris was correlated with vanadate to vanadyl reduction and its accumulation in the growth medium. In two selected strains, JC100/9 and JC100/15, protein kinase A activity was much higher in comparison to the wild type strain even without vanadate addition to the growth medium. Moreover, in the presence of vanadate, protein kinase A activity was significantly increased in the wild type and the vanadate-resistant JC100/1 and JC100/3 strains. It was also found that phosphorylation of a 40 kDa protein associated with ribosomes occured in all vanadate-resistant strains from the logarithmic, while in the wild type strain from the stationary growth phase. From the presented results it can be concluded that a protein kinase A signalling pathway(s) might be involved in the mechanism of P. pastoris vanadate resistance. The results also indicate a possible role of the 40 kDa protein in protection of P. pastoris against vanadate toxicity. PMID- 12545203 TI - Up-regulation of spermidine/spermine N1-acetyltransferase (SSAT) expression is a part of proliferative but not anabolic response of mouse kidney. AB - A differential expression pattern of spermidine/spermine N(1)-acetyltransferase (SSAT), the enzyme critical to proper homeostasis of cellular polyamines, is reported in mouse kidney undergoing hyperplasia and hypertrophy. We have shown that SSAT activity and SSAT mRNA are significantly induced by antifolate CB 3717 and folate that evoke a drug-injury-dependent hyperplasia. In contrast, SSAT activity is down-regulated in the testosterone-induced hypertrophic kidney, while SSAT mRNA is positively controlled by this androgen. Catecholamine depletion evoked by reserpine drastically decreases the folate-induced activity of S adenosylmethionine decarboxylase (AdoMetDC), which limits polyamine biosynthesis, but has no effect on SSAT activity augmented by CB 3717. Our results document that the increased SSAT expression solely accompanies the proliferative response of mouse kidney, and suggest the importance of post-transcriptional regulation to the control of SSAT activity in both hyperplastic and hypertrophic experimental models. PMID- 12545204 TI - IGF-I: from diagnostic to triple-helix gene therapy of solid tumors. AB - Alterations in the expression of growth factors and their receptors are associated with the growth and development of human tumors. One such growth factor is IGF-I (insulin-like growth factor I ), a 70-amino-acid polypeptide expressed in many tissues, including brain. IGF-I is also expressed at high levels in some nervous system-derived tumors, especially in glioblastoma. When using IGF-I as a diagnostic marker, 17 different tumors are considered as expressing the IGF-I gene. Malignant glioma, the most common human brain cancer, is usually fatal. Average survival is less than one year. Our strategy of gene therapy for the treatment of gliomas and other solid tumors is based on: 1) diagnostic using IGF-I gene expression as a differential marker, and 2) application of "triple-helix anti-IGF-I" therapy. In the latter approach, tumor cells are transfected with a vector, which encodes an oligoribonucleotide--an RNA strand containing oligopurine sequence which might be capable of forming a triple helix with an oligopurine and/or oligopyrimidine sequence of the promotor of IGF I gene (RNA-IGF-I DNA triple helix). Human tumor cells transfected in vitro become down-regulated in the production of IGF-I and present immunogenic (MHC-I and B7 expression) and apoptotic characteristics. Similar results were obtained when IGF-I antisense strategy was applied. In both strategies the transfected cells reimplanted in vivo lose tumorigenicity and elicit tumor specific immunity which leads to elimination of established tumors. PMID- 12545205 TI - Carbohydrate moieties of N-cadherin from human melanoma cell lines. AB - Expression of N-cadherin an adhesion molecule of the cadherin family, in tumor cells is associated with their increased invasive potential. Many studies suggested the role of N-linked oligosaccharides as important factors that contribute to metastasis by influencing tumor cell invasion and adhesion. N cadherin is a heavily glycosylated protein. We have analysed the carbohydrate profile of this protein synthesized in human melanoma cell lines: WM35 from the primary tumor site and WM239, WM9, and A375 from different metastatic sites. N cadherin was immunoprecipitated with anti-human N-cadherin polyclonal antibodies. Characterisation of its carbohydrate moieties was carried out by SDS/PAGE electrophoresis and blotting, followed by immunochemical identification of the N cadherin polypeptides and analysis of their glycans using highly specific digoxigenin or biotin labelled lectins. The positive reaction of N-cadherin from the WM35 cell line with Galanthus nivalis agglutinin (GNA), Datura stramonium agglutinin (DSA) and Sambucus nigra agglutinin (SNA) indicated the presence of high-mannose type glycans and biantennary complex type oligosaccharides with alpha2-6 linked sialic acid. N-cadherin from WM239, WM9, and A375 cell lines gave a positive reaction with Phaseolus vulgaris leukoagglutinin (L-PHA) and lotus Tetragonolobus purpureas agglutinin (LTA). This indicated the presence of tri- or tetra-antennary complex type glycans with alpha-fucose. In addition, N-cadherin from WM9 (lymphomodus metastatic site) and A375 (solid tumor metastatic site) contained complex type chains with alpha2-3 sialic acid (positive reaction with Maackia amurensis agglutinin--MAA). The results demonstrated that N-glycans of N cadherin are altered in metastatic melanomas in a way characteristic for invasive tumor cells. PMID- 12545206 TI - Binding of the basic fibroblast growth factor (bFGF) by soluble components of human umbilical cord. AB - Pre-eclampsia, the most common pregnancy associated syndrome, is connected with remodelling of extracellular matrix of the umbilical cord tissues. Since the fibroblast growth factor (FGF) is known to be a stimulator of collagen and glycosaminoglycan biosynthesis, one may expect that it plays an important role in such a remodelling. Studies performed on the umbilical cords of 10 control and 10 pre-eclamptic newborns demonstrated that both the umbilical cord arterial wall and Wharton's jelly contain FGF mainly in complexes with the components of different molecular mass. Pre-eclampsia is associated with a decrease of endogenous FGF-binding by soluble high molecular mass components of the umbilical cord. It is suggested that FGF released from these complexes may be actively bound by fibroblasts of the umbilical cord, stimulating them to produce collagen and sulphated glycosaminoglycans. PMID- 12545207 TI - Inhibition of proteolytic processing of adenoviral proteins by epsilon aminocaproic acid and ambenum in adenovirus-infected cells. AB - Maturation of adenovirus particles is markedly affected by proteolytic processing. The possibility for blocking the conversion of precursor structural core protein (preVII) into mature structure protein VII by officinal drugs epsilon-aminocaproic acid and ambenum has been demonstrated in Hep-2 cells infected with adenovirus. Proteolytic processing may be regarded as one of the targets for inhibiting adenovirus reproduction. PMID- 12545208 TI - Molecular cloning and sequencing of rabbit presenilin-1 cDNA fragment. AB - Molecular cloning and sequencing of a cDNA encoding rabbit presenilin-1 (Ps1) fragment was performed by reverse transcription polymerase chain reaction (RT PCR) using primers: 5'-GGA TGA GCA GCT AAT CTA TAC C-3' and 5'-TCC ATT CAG GGA GGT ACT TGA TA-3'. The cDNA fragment revealed 402 nucleotides. The sequence was well conserved and found to be 91, 90, 88, 87 and 78% homologous to that of human, lemur, rat, mouse and chicken, respectively. The cDNA translated into a 130 amino-acid protein fragment. The deduced amino-acid sequence was also well conserved in various species and exhibited 98% similarities with those of rat, lemur and human homologues. However, differences were noticed at residues 145, 168 and 212. This cDNA fragment is quite significant because it is the most conserved portion of Ps1 in various animals and encodes four transmembrane regions (TM2, 3, 4, 5) as defined in human Ps1. Moreover, it includes more than 50% of the sites at which substitutions have been reported in familial Alzheimer's disease (FAD). Therefore, it is suggested that the rabbit can be used as an experimental model for future studies on Ps1 and its physiological functions to work out possible pathways leading to FAD linked neurodegeneration. PMID- 12545209 TI - High performance liquid chromatography and photodiode array detection of ferulic acid in Rubus protoplasts elicited by O-glycans from Fusarium sp. M7-1. AB - So far only little data have been available concerning the eliciting capacity of well defined glycan molecules isolated from plant pathogens. This study brings new information about changes in plant cells caused by fungal pathogens. Sugar fractions derived from glycoproteins isolated from the fungus Fusarium sp. M7-1 have been tested here as signaling molecules. The ability of three O-glycan fractions (named in this work inducer I, II, III) to trigger responses in Rubus protoplasts has been examined. It was found that inducer III was the most efficient as it elicited changes in the levels of phenylpropanoid pathway intermediates in relation to phenylalanine-ammonia lyase (PAL) activation. PMID- 12545210 TI - The mechanism of azide activation of polyphenol oxidase II from tobacco. AB - So far, azide has been consistently reported to act as an inhibitor of metal enzymes, especially copper proteins. The present work shows that azide can also act as an activator of polyphenol oxidase II (PPO II) from tobacco leaves. From the square-wave voltammetry of native PPO II, peroxide-PPO II complex and azide PPO II complex, the reduction of nitro blue tetrazolium by the enzymes and activation of PPO II by peroxide it follows that the binding of azide to PPO II induces the formation of CuO(2)(2-)Cu in the active site of PPO II from CuO(2)( )Cu in native PPO II. The reason for azide acting as an activator can be attributed to azide complexing with PPO II, thus inducing the formation of CuO(2)(2-)Cu, which is the active site of the peroxide-PPO II complex in which peroxide plays the role of activator. PMID- 12545211 TI - RFLP analysis of 1-aminocyclopropane-1-carboxylate synthase ACC2 and ACC4 genes from Polish cultivars of tomato. AB - An important trait of tomato is the rate of fruit ripening, strongly dependent on ethylene production. The ripening-related ethylene synthesis in tomato is controlled mainly by 1-aminocyclopropane-1-carboxylate synthase LE-ACS2 and LE ACS4 isoenzymes (Rottmann et al., 1991, J. Mol. Biol. 222: 937; Lincoln et al., 1993, J. Biol. Chem. 268: 19422; Barry et al., 2000, Plant Physiol. 123: 979). In spite of numerous reports on the LE-ACS2 and LE-ACS4 gene expression, only ones considered the genomic organisation each of these genes (Rottmann et al., 1991; Lincoln et al., 1993) reported one copy of each of these genes in tomato cv VF36. In this article we suggest that the genomic organisation of LE-ACS2 and LE-ACSS4 genes may depend on tomato cultivars and may differ from that described by the above authors. The results of Southern analyses of genomic DNAs from 17-day old seedlings (cultivars Jaga, Halicz, Betalux, New Yorker) imply that the genomic organisation of LE-ACS2 and LE-ACS4 genes in Polish cultivars differs from that reported for cv VF36. PMID- 12545212 TI - Lipopolysaccharides from Mesorhizobium huakuii and Mesorhizobium ciceri: chemical and immunological comparative data. AB - Lipopolysaccharides of two Mesorhizobium species of different host specificity were compared: M. huakuii and M. ciceri. M. huakuii sp. was represented by five strains with special consideration of M. huakuii IFO 15243(T). SDS/PAGE profiles revealed that all M. huakuii LPS preparations contained low molecular mass fractions (LPS-II) of the same molecular size. All of lipopolysaccharides contained high molecular mass fractions (LPS-I). However, the high molecular mass fraction from each strain possessed an individual molecular size distribution pattern. The crossreactivity of blotted lipopolysaccharides with rabbit polyclonal antibodies against Mesorhizobium huakuii IFO 15243(T) whole bacteria indicated the presence of common epitope(s) within the investigated Mesorhizobium huakuii strains. Moreover, LPS from M. huakuii S52 also reacted with anti M. ciceri HAMBI 1750 serum showing that there are epitopes common for different mesorhizobial species. LPS isolated from Mesorhizobium huakuii strain IFO 15243(T) contained neutral sugars: L-6-deoxytalose, L-rhamnose, D-galactose and D glucose, aminosugars:D-quinovosamine, D-glucosamine, D-2,3-diamino-2,3 dideoxyglucose and D-galacturonic and D-glucuronic acids. In the LPS preparation, fatty acids typical for Mesorhizobium strains were detected. 3-Hydroxydodecanoic, 3-hydroxy-iso-tridecanoic, 3-hydroxyeicosanoic, 3-hydroxyheneicosanoic and 3 hydroxydocosenoic acids were the major amide linked fatty acids, while iso heptadecanoic, eicosanoic, docosenoic, as well as 27-hydroxyoctacosanoic and 27 oxooctacosanoic acids were the dominant ester linked fatty residues. PMID- 12545214 TI - [Protein phosphatase 2A: its structure, function and activity regulation]. AB - Protein phosphatase 2A (PP2A) is a major kind of serine/threonine protein phosphatase in eukaryotic cells. PP2A has many subunits and isozymes encoded by different genes to form many different PP2A holoenzymes. PP2A plays important roles in regulation of cell cycle, signal transduction, cell differentiation, and transformation. Its activity is related to several diseases, including neurodegenerative diseases and cancer. The regulation of PP2A activity is accomplished by its subunits' expression, cellular localization, phosphorylation and methylation. The second messenger, ceramide, also plays an important role in the regulation of PP2A activity, as well as a plenty of low molecular mass inhibitors of PP2A and other protein phosphatases. PMID- 12545216 TI - Excessive expression of the scavenger receptor class A type I can significantly affect the serum lipids. AB - Scavenger receptor (SR) is characterized by its ability to bind negatively charged macromolecules, particularly the modified lipoproteins that are pertinent to the development of vascular disease. To determine the role of excessive scavenger receptor A in the serum lipoprotein metabolism, transgenic mice lines with mouse scavenger receptor A gene type I (SR-AI) under the control of human SR AI enhancer and metallothionein gene promotor were established. After zinc induction, the expression of SR-AI in transgenic mice was a little higher than the controls, but the serum lipids levels were significantly different from the controls, especially the cholesterol. These results demonstrated that overexpression of SR-AI significantly affected the serum lipids levels. PMID- 12545215 TI - The interactions of LIM protein KyoT with polycomb proteins. AB - LIM domain protein KyoT interacts with transcription factor RBP-J and modulates Notch signaling pathway. To study the function of KyoT, yeast two-hybrid was performed and proteins interacting with KyoT2 including Ring1 and hPc2 (human polycomb 2) that belong to PcG(polycomb group) were analyzed. The interactions between Ring1, hPc2, and KyoT2 were confirmed by changing vectors in two-hybrid analysis. Their interactions in vitro were confirmed by GST-pull down assay. Two hybrid assay showed that the LIM domain of KyoT was responsible for interacting with the full-length Ring1 and hPc2. Moreover, the LIM domain also interacted with the C-terminus of hPc2. Interactions of KyoT2 with Ring1 and hPc2 suggested that PcG family might be involved in Notch signaling pathway. PMID- 12545217 TI - Influence of A7-B7 disulfide bond deletion on the refolding and structure of proinsulin. AB - To probe the role of [A7-B7] disulfide bond in the structure and folding of proinsulin, [A7, B7Ser]-proinsulin was prepared. The differences in in vitro refolding, oxidation of free thiol groups, circular dichroism (CD) spectra, antibody and receptor binding assays, and sensitivity to tryptic digestion between the mutant and the wild type proteins were studied. The deletion of [ A7 B7] disulfide bond in proinsulin resulted in a significant decrease of alpha- helix content of the molecule and a great increase in sensitivity to tryptic digestion. The [A7-B7] disulfide bond deleted proinsulin showed a great loss of its receptor binding activity. The in vitro refolding study indicated that the r ate of the oxidation of free thiol groups in the mutant was a little slower at t he later stage as compared to the native molecule, but the deletion of [A7-B7 ] disulfide bond had little effect on the refolding yield. A possible proinsulin folding pathway way was proposed. During the folding, the intra-A chain disulfide bond forms first and very fast. The formation of the [A20-B19] disulfide bond is more crucial than [A7-B7] disulfide bond and forms very likely earlier than the latter. PMID- 12545218 TI - [Enhancement of the production of SAM by overexpression of SAM synthetase in Pichia pastoris]. AB - S-Adenosyl-L-methionine(SAM) is an important metabolic intermediate in the metabolic flux of sulphur. SAM is involved in three key metabolic pathways: transmethylation, transsulfuration and polyamine synthesis. As a potential therapeutic agent, SAM is being used as over the counter drug and nutrient supplement. An expression vector, harboring SAM synthetase 2 gene from S. cerevisiae and regulated by the glyceraldehyde-3-phosphate dehydrogenase gene promoter P(GAP), was transformed into GS115 strain of P. pastoris. Through zeocin resistance and expression screening, a recombinant strain was obtained that had high SAM yield and the fermentation conditions were optimized. The results showed that carbon source, nitrogen source, pH and dissolved oxygen had significant effects on the accumulation of SAM. The SAM production of the recombinant cells reached 2.49 g/L after fermentation for three days under the optimized conditions. The present studies show that fermentation of recombinant P. pastoris strain, expressing heterologous SAM synthetase gene, may be a promising approach for the production of SAM. PMID- 12545219 TI - [Effect of N-terminal deletion on biological activity of vascular endothelial cell growth inhibitor]. AB - Vascular endothelial cell growth inhibitor (VEGI) is a novel cytokine which belongs to the TNF superfamily. It can inhibit the proliferation of endothelial cells and neovascularization. However, little is known about the structure function relationship of VEGI. In order to study the effect of the N-terminus of VEGI on biological properties, the sequence alignment among VEGI and TNF superfamily members based on structure knowledge was done, and then two truncated forms of VEGI were constructed, in which 43 and 51 amino acids from N-terminus were deleted and named VEGI(131) and VEGI(123). Recombinant proteins were generated from E. coli. The expression rates were 25.2% (VEGI(131)) and 27.8% (VEGI(123)) of total bacterial proteins. After purification the purity reached 92.5% (VEGI(131)) and 91.6% (VEGI(123)). VEGI(131) showed significant inhibitory effect on growth of human umbilical vein endothelial cells (HUVEC), IC(50) of VEGI(131) being 35 mg/L. Under the same conditions, IC(50) of VEGI(151) (the wild type of VEGI) was 27 mg/L, but VEGI(123) showed no inhibitory effect. On chick choriallantic membrane (CAM) assay, VEGI(151) markedly reduced the number of main vessels, and VEGI131 decreased capillary number, while the effect of VEGI(123) was almost the same as control. These results suggest that the first 43 amino acids from N-terminus of VEGI have no significant effect on biological activity, but the amino acids 44-51 at N-terminus are required for full biological activity. PMID- 12545220 TI - [Expression and characterization of Kringle 1-4.5 domains of human plasminogen]. AB - The cDNA encoding Kringle 1-4 and part of Kringle 5 domains of human plasminogen (K1-4.5), obtained from HepG2 by RT-PCR, was cloned into expression vector pHIL S1. The recombinant plasmid pHIL-K1-4.5 was transformed into Pichi pastoris GS115 and the recombinant yeast was induced to express the recombinant proteins by methanol. The expressed proteins were purified by lysine affinity chromatography to a purity of 95%. The recombinant K1-4.5 inhibited the growth of bovine capillary endothelial cells (BAEC) stimulated by the basic fibroblast growth factor (bFGF), in a dosage-dependent manner with a half maximal concentration of 2 mg/L. rhK1-4.5 also inhibited 40% of the BAEC migration stimulated by bFGF in the concentration of 1 mg/L. PMID- 12545221 TI - Cloning and sequence analysis of tumor-associated gene hMMTAG2 from human multiple myeloma cell line ARH-77. AB - In order to look for the tumor-associated genes from human multiple myeloma (MM), a cDNA library of human multiple myeloma cell line ARH-77 was constructed with eukaryote expression vector pcDNA3.1(+). The length of inserted fragments in library was 1.2 kb in average. All clones in cDNA library were transferred in situ to nylon membrane, which was divided into eight equal parts (A-H) and cultured in LB medium to set up gene pools. The plasmids in cDNA library and in gene pools were extracted and NIH/3T3 cells were transfected respectively. By G418 screening and colonies counting, gene pool A was chosen for the second cycle transfection. After several cycles, a clone, A62-17, was obtained, which had significant transforming ability. The length of this clone was 993 bp. The RACE technique was used for rapid amplification of A62-17 5'-end. The full length of this sequence has 1300 bp and was named as hMMTAG2 gene. hMMTAG2 consists of 8 exons and codes for a polypeptide of 263 amino acids (the accession number in GenBank: AY137773). It was located at chromosome 1q42.13. hMMTAG2 had same transforming activities in NIH/3T3 cells as the clone A62-17, and the number of transformant foci was 6 folds more than the blank vector pcDNA3.1(+). The analysis of bioinformatics revealed that hMMTAG2 had many phosphorylation sites for several protein kinases, N-myristoylation sites and nuclear localization signals, so it may be a signal molecule in the nucleus. PMID- 12545222 TI - [Mutation of arginines near the active site Cys124 of human dual-specificity phosphatase and its effect on the enzymatic activity]. AB - To study the effect of three positively charged arginine residues near the active site Cys(124) of the human dual-specific phosphatase on the catalytic function, six VHR mutants R(125)L, R(130)L, R(130)K, R(130)L/S(131)A, R(158)K and R(158)L were obtained using QuikChange site-directed mutagenesis method. The recombinant plasmids containing mutant genes were transformed into the Escherichia coli strain BL21(DE3), and the expressed proteins were found to be water soluble after the induction of IPTG. The proteins with purity greater than 90% were obtained using Ni(2+) chelating affinity chromatography. The measurement of the steady state kinetic parameters and arsenate inhibition constants K(i) of the enzyme and their mutants showed that the k(cat)/K(m) values of Arg(130) and Arg(158) mutants decreased, and K(i) values increased obviously compared with those of the wild enzyme. These results indicated that Arg(130) and Arg(158) were necessary for the enzymatic activity, and were probably related to the binding with the negatively charged phosphate group of the substrate. In addition, the slight difference for the k(cat) values between the R(130)L and R(130)L/S(131)A mutants suggested that Arg(130) mutation disrupted the hydrogen bond between Ser(131) and Cys(124). Furthermore, the arsenate binding affinity for R(125)L, R(130)L and R(158)L mutants was decreased, suggesting that positive charges in the side chains of these three arginine residues may be helpful for the binding of the enzyme to the substrate. PMID- 12545223 TI - [Influence of signal peptide sequences on the expression of heterogeneous proteins in Pichia pastoris]. AB - Pichia pastoris has been developed to be a very efficient expression host for the heterogeneous proteins since its alcohol promoter was isolated and cloned, and its transformation technique was established. For further improving the secretion expression of heterogeneous proteins, in this research, the signal sequences were studied. At first, the Saccharomyces cerevisiae mating factor alpha prepro-leader sequence was synthesized using successive PCR and designated as MF4I. Then, ten different signal sequences were constructed by adding the N-terminal residues of Pichia pastoris Aox1 protein to the N-terminal of the MF4I. These ten signal sequences were used for directing phytase gene secretion in Pichia pastoris, the secretion of phytase were increased in Pichia pastoris strains containing new signal sequence. Among these strains, the phytase secretion was highest in strain contain signal sequence added with A, I, P three Aox1 N-terminal residues; the phytase secretion of Pichia pastoris was 90 mg/L in flake. The secretion was six fold of that with original Saccharomyces cerevisiae mating factor alpha prepro leader sequence. In addition, insert of ten residues E E A E A E A E P K can further increase the phytase secretion by 35%, the secretion reach 120 mg/L. PMID- 12545224 TI - [The effects of PTEN gene on migration and FAK phosphorylation of SMMC-7721 human hepatocarcinoma cell line]. AB - PTEN is a major tumor suppressor gene that encodes a dual-specificity phosphatase with high sequence similarity to the cytoskeletal protein tensin. PTEN may be involved in the formation and disassembly of focal adhesion and affect cell migration. In the present study, PTEN expression plasmid was constructed and transfected into the hepatoma cell line SMMC-7721 to analyze the alterations of cell motility and FAK tyrosine phosphorylation. It was observed that the overexpression of PTEN gene significantly inhibited cell motility on extracellular matrix (Fn), and the cell migration on fibronectin was reduced by 35%. Similarly, at 30-min and 60-min, the cell spreading on Fn but not on polylysine was inhibited by 29% and 26% respectively. The data obtained from immunoprecipitation and immunoblotting analyses showed that the overexpression of PTEN did not affect FAK expression but resulted in a decrease in FAK tyrosine phosphorylation. The level of FAK phosphorylation was inversely correlated with the level of PTEN protein in three cell lines. It was also found that the overexpression of PTEN led to growth inhibition, with the number of cells in S phase reduced by 16%. These results indicate that PTEN exerts its tumor suppressive effects on hepatocellular carcinoma cells through the inhibition of cell motility and cell cycle progression. PMID- 12545225 TI - [Secretory Expression of the Superactive [Lys(17,18),Glu(21)]-Glucagon in E. coli]. AB - One of the most important findings in structure-function studies on glucagon by means of chemical synthesis is the discovery that [Lys(17,18),Glu(21)]-glucagon had higher biological activity than native glucagon. This mutant of glucagon was called superactive glucagon (SA-glucagon). In the present work, the possibility to obtain SA-glucagon by means of genetic engineering was studied. The gene of SA glucagon (SAG) was obtained by PCR from a constructed recombinant glucagon plasmid, pAGluT. A secretory expression vector harboring SAG, pBLSG7, containing P(L) promoter and the gene of phoA signal peptide was constructed. In expression studies after transformation of pBLSG7 into E. coli BL21, it was found that the expression yield of SA-glucagon reached 3.65 mg/L(A(600)=1), about 19.5% of total proteins in the culture medium under shaken flask conditions. In addition, the influence of induction temperature and of E. coli strain on the expression yield of SA-glucagon was also studied. PMID- 12545226 TI - [Cloning, expression of the lectin-EGF domain of P-selectin, and preparation of its monoclonal antibody]. AB - To prepare monoclonal antibody specific to P-selectin lectin-EGF domain, the gene for lectin-EGF domain of P-selectin L-EGF was amplified from normal human platelets by RT-PCR, then was cloned into prokaryotic vector pET42b(+). The recombinant plasmid was transformed into E. coli DH5 alpha strain for further screening and characterization, and was expressed in E. coli BL21 strain. Expressed protein was purified by chromatography on a Ni(2+)-NTA superflow agarose column and eluted with pH 8.0-4.5 urea gradient. Then the mAb anti-lectin EGF was prepared with classical hybridoma technique, and 3 hybridoma cell lines (B10, F3 and H5) were obtained with Ig subclasses of these mAbs were IgG(2), IgG(1), and IgG(3) respectively, and their light chains were all kappa chain. Immuofluorescence and FACS assays demonstrated that mAbs could specifically recognize P-selectin expressed on ECV (endothelial cell line) stimulated by LPS. Meanwhile, the role of mAbs to P-selectin lectin-EGF domain was studied, and it was proved that the mAbs markedly inhibited adhesion between platelets and neutrophils in vitro. These monoclonal antibodies can specifically recognize the natural P-selectin and markedly inhibit adhesion between platelets and neutrophils in vitro. PMID- 12545227 TI - Synthesis of osteogenic growth peptide and its synergetic effect with granulocyte colony stimulating factor. AB - Osteogenic growth peptide (OGP) has been synthesized through Fmoc solid phase synthesis procedure. The purity of synthetic OGP (sOGP) is over 98.6% identified by HPLC, the amino acid sequence and electro-spray mass spectroscopy are consistent with theoretical values. The synergetic effect of sOGP with recombinant human granulocyte-colony stimulating factor (rhG-CSF) on the hematopoiesis was investigated in normal mice. To assess the synergy of sOGP with rhG-CSF, two schemes were designed. In one scheme rhG-CSF was used at the last 8 days of a 13-day treatment with sOGP, while in the other one both cytokines were given concurrently for 10 days [sOGP, 0.5 nmol/day (mouse); rhG-CSF, 2 microg/day (mouse)]. Both schemes showed that sOGP remarkably synergized with rhG-CSF on increment of white blood cell number and lymphocyte number in peripheral blood without any change of red blood cell and platelet counts. Quantitative differential analysis of bone marrow and histological examination of the spleen and sternum showed that sOGP plus rhG-CSF did not cause abnormal hyperplasia, so sOGP is a very hopeful new drug to improve the effectiveness of clinical used rhG CSF. PMID- 12545228 TI - [Cloning, expression and alternative splicing of a novel isoform of human TCP11b gene]. AB - Based on homology analysis and RT-PCR, a novel isoform of human TCP11 gene was isolated. It encodes a 503 amino acid protein that is highly homologous to the mouse 566 amino acid protein Tcp-11. Tcp-11 is important to sperm function because it may be the receptor of fertilization promoting peptide (FPP). Complicated alternative splicing was found to exist between TCP11a and TCP11b genes. The gene has been mapped to human chromosome band 6p21 by fluorescence in situ hybridization. Results of Northern blot and RT-PCR analysis of RNA extracted from human tissues revealed that the gene was expressed in fertile adult testes only, but neither in azoospermic patient testes, fetal testes nor in other human tissues. Our results suggest that TCP11b gene may be important to human sperm function and male fertility. PMID- 12545229 TI - [Purification of a Storage Protein from Aphis craccivora Koch and Its Subunit Characterization]. AB - By using of DE52 Cellulose, Sephadex G-150 and FPLC Q Sepharose chromatography, a storage protein (SP) was purified from Aphis craccivora Koch and its subunit was characterized. Its molecular weight was about 60 kD as determined by SDS-PAGE analysis under both reducing and non-reducing conditions, and its pI was about 5.0. It was a glycoprotein. The protein accumulated in larval hemolymph, and decreased when host turned to adult, and could be detected in adults in very low concentration. According to the molecular weight, amino acids composition, and its dynamic alteration of concentrations, the protein should be a persisting storage protein of hemimetabolous insects. PMID- 12545230 TI - [The relation between translation speed and protein secondary structure]. AB - Based on the statistical analysis of 119 human and 92 E. coli proteins it was found that for both human and E. coli, the mRNA sequences consisting of tri-codon and tetra-codon with high translation speed preferably code for alpha helices more than for coils. For beta strand, the preference/avoidance oscillates with the translation speed. Moreover, the non-homogeneous usages of tri-codon and tetra-codon with different translation speeds in a given secondary structure have also been found. These results cannot be simply explained by the effect of stochastic fluctuation. PMID- 12545231 TI - Dimensional measures of psychopathology. The probability of being classified with a psychiatric disorder using empirically derived symptom scales. AB - OBJECTIVES: We sought to develop a ranking scheme that assigns a probability of having one of four psychiatric disorders to children based on their scores on a symptom scale. We then estimated the impact of each scale symptom on the prevalence of the disorder in the population. METHOD: Logistic regressions were specified for ADHD, ODD, depressive, and conduct disorders using all the individual symptoms in the pertinent scale as predictors. Individual fitted values from the regression function then served as a probability scale measure. We combined the prevalence and influence of each scale symptom to calculate its overall impact on the prevalence of the disorder. RESULTS: Probability distributions had a wide range of values and discriminated between cases and non cases. Those having a disorder were consistently associated with higher probabilities in the scale. The estimated probability corresponds to the empiric prevalence of the diagnosis in a group of persons sharing the same estimated probabilities. Symptoms varied on their impact on the prevalence of the disorder. CONCLUSIONS: We recommend the estimated probability of the disorder based on the empirically defined scales as dimensional measures that complement prevalence of the disorder. Different symptoms are identified as targets for screening when selection is based on their impact on the prevalence of the disorder than when selection is based on the strength of the association with the disorder. We recommend using a common nosology with different classification schemes; the categorical definition of the disorder, the probability of having the disorder, and the impact of each symptom in the prevalence. Different measures serve different purposes. PMID- 12545232 TI - Mental health service utilization in the United States. The role of personality factors. AB - BACKGROUND: The aim of this study was to determine the relationship between personality factors and the use of mental health services (past 12 months) among adults in the community. METHOD: Data were drawn from the Midlife Development in the United States Survey (MIDUS), a representative sample of 3,032 adults aged 25 74 in the United States population. Analyses of variance and logistic regression analyses were used to determine the relationship between personality factors and mental health service utilization, in the presence and absence of mental disorders, during the past 12 months. RESULTS: Neuroticism [OR = 1.5 (1.2, 1.9)] was associated with significantly increased likelihood of mental health service utilization among adults in the community. Conscientiousness [OR = 0.7 (0.5, 0.9)] and extraversion [OR = 0.7 (0.5, 0.98)], in contrast, were associated with decreased likelihood of use of mental health services. Among adults with mental disorders, conscientiousness [OR = 0.5 (0.3, 0.8)] was associated with decreased odds of mental health service utilization. Neuroticism [OR = 1.8 (1.3, 2.4)] was associated with increased likelihood of service use among those who did not meet criteria for common mental disorders. CONCLUSIONS: These findings are the first to document a significant association between personality factors and the use of mental health services among adults in the general population. Our results highlight new ways in which personality may influence mental health in the community. This information may be useful in identifying those with unmet need for mental health treatment and developing more effective interventions for those at risk for common mental disorders. Replication of these findings is needed. PMID- 12545233 TI - Prevalence and risk factors for depression in a rural setting. Results from the North Wales arm of the ODIN project. AB - BACKGROUND: Depressive disorders are common, and result in high individual and societal costs. The majority of research assessing depression has occurred in urban areas. There is a paucity of research examining the prevalence of and risk factors for depression in rural general practice. AIM: The aim of this study was to examine the prevalence of and risk factors for depression in a rural area of North Wales in the context of a large multi-centre European study. METHOD: One thousand nine hundred and ninety-nine people randomly selected from a health authority database underwent a two-phase screening method to identify depression. The first phase involved patients completing a self-rating postal questionnaire (the Beck Depression Inventory or BDI). In the second phase, those scoring above cut-off underwent detailed diagnostic interview (Schedules of Clinical Assessment in Neuropsychiatry or SCAN). The SCAN diagnostic interview can generate either DSM-IV or ICD-10 diagnoses; the DSM-IV classification system was used here. RESULTS: One thousand two hundred and thirty-nine (63 %) people responded to the initial screening questionnaire. The prevalence rate for all DSM-IV depressive disorders was calculated to be 6.1 % (95 % CI 4.1, 9.0) whereas the prevalence rate for DSM-IV major depressive disorder was 5.1 % (CI 3.37, 7.66). Multivariate analysis indicated that several variables were related to BDI caseness, including gender, employment status, social support and negative life events. CONCLUSION: Levels of reported depression are relatively low in North Wales compared to those observed in a neighbouring urban area using comparable data collection methods. A number of factors traditionally associated with increased risk from depression were predictive of scoring above cut-off on the BDI in a rural North Wales sample. PMID- 12545234 TI - Childhood Experience of Care and Abuse Questionnaire (CECA.Q). Validation of a screening instrument for childhood adversity in clinical populations. AB - BACKGROUND: Interview measures for investigating adverse childhood experiences, such as the Childhood Experience of Care and Abuse (CECA) instrument, are comprehensive and can be lengthy and time-consuming. A questionnaire version of the CECA (CECA.Q) has been developed which could allow for screening of individuals in research settings. This would enable researchers to identify individuals with adverse early experiences who might benefit from an in-depth interview. This paper aims to validate the CECA.Q against the CECA interview in a clinical population. METHODS: One hundred and eight patients attending an affective disorders service were assessed using both the CECA interview and questionnaire measures. A follow-up sample was recruited 3 years later and sent the questionnaire. The questionnaire was also compared with the established Parental Bonding Instrument (PBI). RESULTS: Agreement between ratings on the interview and questionnaire were high. Scales measuring antipathy and neglect also correlated highly with the PBI. The follow-up sample revealed the questionnaire to have a high degree of reliability over a long period of time. CONCLUSIONS: The CECA.Q appears to be a reliable and valid measure which can be used in research on clinical populations to screen for individuals who have experienced severe adversity in childhood. PMID- 12545235 TI - Long-term consequences of unrecognised PTSD in general outpatient psychiatry. AB - BACKGROUND: Although a number of studies have shown a high prevalence of PTSD in the population, the diagnosis has hardly been recognised in general psychiatric practice. This raises two important questions. How widespread is extreme trauma and PTSD in the general psychiatry population? How does the long-term outcome among patients with PTSD differ from that of other psychiatric patients? The present study examines a psychiatry outpatient population in which none of the patients have received the primary diagnosis of PTSD. METHOD: The 233 patients from four ethnic groups who, during a period of 1 year, visited a psychiatric outpatient clinic were asked, 3-4 years later, to complete a questionnaire that included the Self-rating Inventory for Posttraumatic Stress Disorder. Patients were divided into probable PTSD and non-PTSD. Subjects with probable PTSD were compared with non-PTSD, regarding outcome at follow-up. RESULTS: The response rate was 56%. Fifty-three patients fulfilled the instrument's diagnostic criteria for PTSD, whereas 62 did not. In the foreign-born groups PTSD patients constituted a majority (53-69 %) compared to 29% of Swedish-born patients. Although the clinic's original diagnoses and assessment of immediate treatment outcome did not differ between the two groups, PTSD patients did not report improvement to the same extent and had poorer self-rated health at follow-up (p < 0.001) as well as lower participation in working life. CONCLUSIONS: A high proportion of PTSD and poorer outcome for PTSD patients compared to other psychiatric patients warrants a primary focus upon PTSD in outpatient psychiatry. PMID- 12545236 TI - First-time hospitalisation for depressive and/or anxiety disorders and severe life events. AB - BACKGROUND: It has been suggested that the impact of antecedent life events differs between first and subsequent episodes of depressive or anxiety disorders. METHOD: We used the Bedford College methodology to determine the presence of severe events and/or major difficulties in two different groups of patients hospitalised for neurotic disorders, and in a control group of healthy subjects from the general population. RESULTS: The experience of severe events and/or major difficulties was significantly higher in patients being hospitalised for the first time than in patients who had been hospitalised several times, or in the control group. This difference was observed for female patients only. CONCLUSIONS: Two hypotheses are put forward as an explanation of these results. PMID- 12545237 TI - The role of stress in the onset of depressive disorders. A controlled study in a Spanish clinical sample. AB - BACKGROUND: We conducted the present study to evaluate the impact of stressful events on the onset of depressive disorders in a Spanish clinical sample, compared to a control group matched for age, sex, civil status and social class. We compared our results with those of other studies carried out with samples that were both clinically and culturally similar to ours. METHOD: Fifty depressed patients that were diagnosed with a depressive episode in the 6 months prior to the interview and 50 healthy controls were included in the study. Both groups were compared on the "Life Events and Difficulties Schedule" (LEDS). RESULTS: Of the depressive patients, 68 % compared to only 18 % of the control individuals experienced at least one provoking agent in the 12 months prior to the onset of the symptoms. The risk of developing a depressive disorder was 9.7 % greater in subjects exposed to such provoking agents. Chronic difficulties are equally important to the genesis of depressive disorders as severe life events. No significant differences were seen between the two diagnostic subgroups of depressed patients in the accumulation of severe events, major difficulties or provoking agents. CONCLUSION: The results support the view that stress is a major factor in the aetiology of depressive disorders. The amount of stress suffered by the patients, however, was less than that found in our healthy sample. Important issues about the model of interaction between stress and depression are discussed. PMID- 12545238 TI - Do patients who self-harm consult their general practitioner soon after hospital discharge? A cohort study. AB - BACKGROUND: Deliberate self-harm (DSH, attempted suicide) is one of the most common reasons for emergency hospital admission in Great Britain. Approximately 20 % of patients repeat self-harm in the 12 months after admission. The GP's role in DSH aftercare and the prevention of repeat episodes is unclear. METHODS: The data were obtained from a 12-month follow-up of a series of 968 consecutive patients from 49 practices who attended an accident and emergency (A&E) department in Bristol or Bath, UK, following an episode of self-harm between 26(th) May 1997 and 1(st) March 1999. RESULTS: Information on consultation patterns were available for 681 (70 %) of the patients. Two hundred and fourteen (31 %) of these consulted their GP in the week following the episode, 360 (53 %) within 4 weeks. Of the subjects, 44 % were discharged directly from A & E; these patients were more likely to consult their GP in the 4 weeks after the episode. Over the 12-month follow-up, 117 (17 %) attended hospital for a repeat episode of DSH; 9 % of these repeats occurred within 1 week and 28 % within 4 weeks of the index event. Although few (2 out of 11 patients--18 %) of those repeating in the first week after the index episode consulted their GP prior to repeating, 50 % (11/22) of those who repeated in the 1-4 weeks after an episode did so. CONCLUSIONS: Consultation patterns indicate that most people who deliberately self-harm consult their GP soon after the episode. This consultation may provide an opportunity for preventing repeat DSH and suicide. PMID- 12545239 TI - On vortex loops and filaments: three examples of numerical predictions of flows containing vortices. AB - Vortex motion plays a dominant role in many flow problems. This article aims at demonstrating some of the characteristic features of vortices with the aid of numerical solutions of the governing equations of fluid mechanics, the Navier Stokes equations. Their discretized forms will first be reviewed briefly. Thereafter three problems of fluid flow involving vortex loops and filaments are discussed. In the first, the time-dependent motion and the mutual interaction of two colliding vortex rings are discussed, predicted in good agreement with experimental observations. The second example shows how vortex rings are generated, move, and interact with each other during the suction stroke in the cylinder of an automotive engine. The numerical results, validated with experimental data, suggest that vortex rings can be used to influence the spreading of the fuel droplets prior to ignition and reduce the fuel consumption. In the third example, it is shown that vortices can also occur in aerodynamic flows over delta wings at angle of attack as well as pipe flows: of particular interest for technical applications of these flows is the situation in which the vortex cores are destroyed, usually referred to as vortex breakdown or bursting. Although reliable breakdown criteria could not be established as yet, the numerical predictions obtained so far are found to agree well with the few experimental data available in the recent literature. PMID- 12545240 TI - The flight of Archaeopteryx. AB - The origin of avian flight is often equated with the phylogeny, ecology, and flying ability of the primitive Jurassic bird, Archaeopteryx. Debate persists about whether it was a terrestrial cursor or a tree dweller. Despite broad acceptance of its arboreal life style from anatomical, phylogenetic, and ecological evidence, a new version of the cursorial model was proposed recently asserting that a running Archaeopteryx could take off from the ground using thrust and sustain flight in the air. However, Archaeopteryx lacked both the powerful flight muscles and complex wing movements necessary for ground takeoff. Here we describe a flight simulation model, which suggests that for Archaeopteryx, takeoff from a perch would have been more efficient and cost effective than from the ground. Archaeopteryx may have made short flights between trees, utilizing a novel method of phugoid gliding. PMID- 12545241 TI - Lowering body temperature induces embryonic diapause during prolonged egg retention in the lizard, Calotes versicolor. AB - The lizard Calotes versicolor delays oviposition of oviductal eggs for as long as 6 months or more under unfavourable conditions. During this period of prolonged egg retention, the growth of oviductal embryos is arrested at stage 34. The present study shows for the first time among reptiles that the "embryonic diapause" is manifested by the gravid females by lowering their body temperature ( T(b)) by 3-5 degrees C during the period of egg retention by mechanism(s) presently unknown. PMID- 12545242 TI - Mole rats ( Spalax ehrenbergi) select bypass burrowing strategies in accordance with obstacle size. AB - Mole rats inhabit extensive individual tunnel systems. Since the energetic cost of burrowing far exceeds that of surface locomotion, excellent orientation ability is crucial. Here we examined whether mole rats can bypass an obstacle (ditch) intersecting an existing tunnel in order to rejoin the two tunnel sections. The mole rats dug two bypass types, depending on the size of the obstacle confronting them: a bypass around the small ditches, parallel and close to the ditch walls; or a bypass beneath the floor of the large ditches. These results demonstrate that the mole rat has the ability to avoid obstacles by digging accurate and energy-conserving bypass tunnels. In order to utilize such a capacity, the mole rat must possess both the means to evaluate the size of the obstacle as well as the ability to perceive its exact position relative to the original tunnel that it will rejoin. Possible mechanisms of orientation that could explain such ability are briefly discussed. PMID- 12545243 TI - Effect of prenatal exposure to hydroxyprogesterone on steroidogenic enzymes in male rats. AB - Pregnant Wistar strain albino rats were administered hydroxyprogesterone and the steroidogenic potential of the testis was analysed in the next generation adult male rats. In utero exposure to supranormal levels (10 mg/kg body weight or 25 mg/kg body weight) of hydroxyprogesterone decreased the activity levels of marker steroidogenic enzymes (3beta-hydroxysteroid dehydrogenase and 17beta hydroxysteroid dehydrogenase) of adult (mature) rat testis. Hydroxyprogesterone mediation in the suppression of rat testicular steroidogenesis is suggested, which can be correlated to the observed decrease in male reproductive potential in mammals exposed to female hormones during embryonic development. PMID- 12545244 TI - Understanding clay minerals with fuzzy mathematics. AB - A long-existing geochemical problem is the lack of a means of chemically defining non-stoichiometric complex minerals such as clays, which have no distinct composition and no clear compositional boundaries. We propose here a novel approach for describing the chemical nature of clay minerals using fuzzy logic. This non-conventional mathematical approach allows us to quantify compositional vagueness in such systems. We show that a clay mineral can be described in terms of how compositionally representative it is, of its own type ("belonging-ness") and to what extent it resembles other types (compositional overlap). Many clay minerals are seen to be far from the ideal, and most minerals are also to a lesser (and sometimes greater) extent, close to types other than their own. This has provided a means of grading such minerals, evaluating how "good" a sample is, and defining the extent of transition to other phases. We have derived here the fundamental methodology for such computations, which could be a framework for the analysis of other complex chemical systems, especially in the fields of geology and metallurgy. PMID- 12545245 TI - Novel mechanistic concepts for the control of leukocyte transmigration: specialization of integrins, chemokines, and junctional molecules. AB - The emigration of leukocytes from the circulation is a critical step during immune surveillance and inflammatory reactions and is governed by a coordinated interplay involving a spectrum of adhesion and signal molecules. As the original multistep model of leukocyte trafficking undergoes continuous revision and refinement, the identification of additional molecules and the emergence of novel concepts for their intricately overlapping functions indicate that this process is still not completely understood. Early studies defining the multistep model described the rapid transition of selectin-mediated leukocyte rolling into integrin-dependent arrest followed by transendothelial diapedesis. It has become apparent that highly specialized chemoattractive cytokines termed chemokines and their heptahelical receptors are involved in the emigration of leukocytes not only by inducing chemotaxis but also by regulating integrins to trigger cell arrest in shear flow. In light of the enormously pleiotropic role of integrins and chemokines in vertebrate biology, this review summarizes and highlights selected aspects of currently evolving concepts refining the multistep model: (a) the differential activation of integrin avidity by chemokines and its implications, (b) the functional specialization of chemokines and their receptors in leukocyte recruitment, and (c) the multilayered molecular "zipper" controlling the completion of diapedesis at interendothelial junctions. PMID- 12545246 TI - The role of VEGF in normal and neoplastic hematopoiesis. AB - VEGF is a secreted growth factor that mediates its biological effects by binding to two transmembrane tyrosine kinase receptors, VEGFR-1 and VEGFR-2. The VEGF/receptor signaling system is involved in the regulation of two fundamental processes in vertebrates: the formation of blood vessels (angiogenesis) and of blood cells (hematopoiesis). Hematopoietic stem cells, capable of giving rise to all blood cell lineages, are often found in clusters with endothelial cells, the key cell type involved in the formation of blood vessels. Despite such proximity of VEGF-responsive cells, hematopoiesis occurs independently of neoangiogenesis in the adult bone marrow, suggesting that VEGF regulates the two processes by different mechanisms. In support of this hypothesis, the recently identified autocrine loop by which VEGF may control hematopoietic stem cell survival and repopulation, is fundamentally different from its paracrine effects regulating angiogenesis. Furthermore, coexpression of VEGF and its receptors, the prerequisite for autocrine loops, is frequently found in lymphomas and myelomas, suggesting that autocrine loops also play a role in hematological malignancies. Several therapeutic strategies blocking VEGF or VEGF-induced signaling are currently being investigated for the treatment of neoplastic diseases. They differ in their potential to interfere with the autocrine or paracrine effector functions of VEGF during angiogenesis, hematopoiesis, and tumor cell proliferation, properties which may ultimately determine their therapeutic potential. PMID- 12545247 TI - Dynamic expression patterns of transforming growth factor-beta(2) and transforming growth factor-beta receptors in experimental glomerulonephritis. AB - Numerous studies have demonstrated the involvement of the transforming growth factor (TGF) isoform beta(1) in the pathogenesis of renal fibroproliferative diseases. Although in vitro studies suggest that TGF-beta(2) is equally potent to TGF-beta(1) in terms of its antimitogenic and fibrogenic effects, much less is known about the regulation of TGF-beta(2) in renal diseases associated with glomerular cell hyperplasia and matrix expansion. Here we investigated the glomerular expression patterns of TGF-beta(2) and of the TGF-beta receptors I, II, and III during the course of rat anti-Thy1.1 nephritis (days 2, 6, 12, and 56), a model characterized by transient mesangial hypercellularity and extracellular matrix accumulation. TGF-beta(2) exhibited dynamic changes in expression. Immunohistochemical double-staining of renal sections revealed that most TGF-beta(2)-positive cells in control glomeruli were podocytes with few TGF beta(2)-positive mesangial cells. This staining pattern could also be observed in human kidney. On day 6 of anti-Thy1.1 nephritis both TGF-beta(2) positive podocytes and mesangial cells were more abundant. By western blot analysis of isolated glomeruli from nephritic rats, protein expression of TGF-beta(2) was upregulated tenfold over control glomeruli, peaking on day 6 of the disease. In cultured rat mesangial cells we found that the TGF-beta(2) and TGF-beta(1) isoforms were equally potent in terms of nuclear accumulation of phosphorylated Smad 2/3, inhibition of DNA synthesis, and induction of beta(1)-integrin and type I collagen protein synthesis. Protein expression of the TGF-beta receptor I was not detected by immunohistochemistry in control glomeruli but was markedly induced in the mesangium on day 6 of nephritis. Mesangial staining for TGF-beta receptors II and III was detected in normal kidneys. Expression of TGF-beta receptor II was strongly enhanced on days 6 and 12 of disease, while TGF-beta receptor III was upregulated only on day 6. In summary, we report marked yet transient upregulation of TGF-beta(2) protein and of TGF-beta receptors I, II, and III in glomerular cells during anti-Thy1.1 nephritis. These results are in keeping with the notion that TGF-beta(2) and its receptors participate in the pathogenesis and/or resolution of this transient form of glomerulonephritis. PMID- 12545248 TI - HLA-A3 restricted mutant ras specific cytotoxic T-lymphocytes induced by vaccination with T-helper epitopes. AB - Cytotoxic T-lymphocytes are one of the most important elements of the antitumor defense. Stimulation of cytotoxic T-lymphocytes outgrowth after immunization with mutant ras peptides is a desired goal since these cells may kill tumor cells in vivo. In this study we tested responding peripheral mononuclear cells from a patient with pancreatic adenocarcinoma who had received intradermal peptide vaccination with a mixture of 17-mer mutant ras peptides and granulocyte macrophage colony-stimulating factor as an adjuvant. Responding peripheral T cells were cloned by limiting dilution and several CD8(+) cytotoxic T lymphocytes, specific for the K- RAS 12-Cys mutation were obtained. By using a panel of nonamer peptides containing the 12-Cys mutation and covering position 4 21 in the ras molecule, the 9-mer peptide which was actually recognized by the cytotoxic T-lymphocytes could be identified. HLA-A*0302 could be identified as the antigen-presenting molecule, and the amino acid sequence of the T-cell epitope carries the previously identified HLA-A*0302 binding motif. The nonamer peptide was contained within the vaccine peptide originally used for intradermal immunization of the patient. The cytotoxic T-lymphocytes were capable of killing target cells expressing HLA-A*0302 that coexpressed the K- RAS 12-Cys mutation after transfection. These data demonstrate that the peptide used for vaccination (17-mer) is processed and presented in vivo, and that generation of cytotoxic T lymphocytes by vaccination with T-helper epitopes may be important for further development of specific immunotherapy of cancer patients. PMID- 12545249 TI - Resolving the composite trait of hypertension into its pharmacogenetic determinants by acute pharmacological modulation of blood pressure regulatory systems. AB - Acute pharmacogenetic analysis was carried out in an intercross F2 population derived from Prague hypertensive-hypertriglyceridemic and Lewis rats. Quantitative trait loci (QTL) mapping was performed for baseline blood pressure (BP) and for BP after blockade of the renin-angiotensin system by losartan, of the sympathetic nervous system (SNS) by pentolinium, and of the nitric oxide system by N(G)-nitro- L-arginine methyl ester. Two significant loci for baseline BP were found on chromosome (Chr) 3 (logarithm of likelihood, LOD, 3.8) and Chr 5 (LOD 3.6), and one suggestive locus on Chr 1 (LOD 2.7). The QTL on Chr 3 persisted after treatment with the three agents while the QTL on Chr 5 and Chr 1 disappeared after pentolinium administration. This suggests independence of the locus on Chr 3 from each acute BP regulatory system examined, whereas the loci on Chr 5 and Chr 1 appeared to be controlled mainly by the SNS. Although not apparent at baseline, a significant locus appeared on Chr 8 (LOD 7.0) after blockade of the SNS, and NO system blockade led to the appearance of a new QTL on Chr 1 (LOD 3.6), indicating the contribution of the inhibited systems to these loci. Pharmacogenetic dissection of the BP trait is a powerful tool to unravel the underlying physiological mechanisms of QTL affecting baseline BP and to identify specific QTL for the response to drugs. This pharmocogenetic approach enabled us to determine the main causative acute BP regulatory systems and should lead to better selection of suitable antihypertensive drugs for individual patients. PMID- 12545250 TI - Elevated methyl-CpG-binding protein 2 expression is acquired during postnatal human brain development and is correlated with alternative polyadenylation. AB - Rett syndrome is caused by mutations in MECP2 and characterized by arrested postnatal neurodevelopment. MECP2 is ubiquitously expressed, but its protein product, methyl-CpG-binding protein 2 (MeCP2), is highly expressed in a subpopulation of cells in the adult brain. Automated quantitation of MeCP2 expression on a human developmental tissue microarray was performed by laser scanning cytometry. A significant correlation between age and MeCP2 level, population heterogeneity, and percentage of MeCP2 high-expressing cells was specifically observed in cerebral but not renal samples. In contrast, an inverse correlation between use of the long 3' UTR of MECP2 and age was observed, suggesting that an acquired switch in polyadenylation is responsible for the elevated MeCP2. Acquired elevated MeCP2 expression in neurons beginning in infancy and progressing through childhood may explain the delayed onset and developmental arrest of Rett syndrome PMID- 12545251 TI - Preliminary results of the new 6F TrapEase inferior vena cava filter. AB - The TrapEase filter is a permanent, symmetric nitinol filter that can be deployed through a low-profile sheath. Although the TrapEase is enjoying an increasing market share of inferior vena cava (IVC) filters, there are still limited clinical follow-up data on its use. This study is a retrospective review of 189 consecutive infrarenal TrapEase filters placed at our institution. The study included 80 men and 109 women, with an average age of 73 years (24-102). The most common indication for filter placement was a contraindication to warfarin. In total, 13% of the patients were treated with warfarin. The filter was successfully deployed in all cases, via the right femoral vein in 57% of the patients, via the left femoral vein in 32%, and via the right internal jugular vein in 11%. Follow-up studies were performed as clinically indicated. During the investigation, there were three cases of IVC thrombosis (1.5%) and one case of retroperitoneal hemorrhage potentially caused by filter placement. Two of the thrombosis cases resulted in serious sequelae. This study also represents the first report of a symptomatic pulmonary embolism (PE) after TrapEase filter placement. While this investigation does demonstrate a low overall complication rate of this new device, it raises the concern of an increase in IVC thrombosis rate. PMID- 12545252 TI - Aortic aneurysm sac shrinkage after endovascular repair is device-dependent: a comparison of Zenith and AneuRx endografts. AB - Aneurysm sac shrinkage after endovascular aneurysm repair (EAR) provides objective evidence of successful aneurysm exclusion and absence of endotension. Attainment of this outcome parameter may be device-dependent. In this study, 169 patients underwent EAR with an AneuRx (n = 118) or Zenith (n = 51) endograft at a single institution. A prospectively maintained database was examined for significant changes in aneurysm sac diameter (> or = 5 mm) on the basis of computed tomography (CT) measurements at 6 and 12 months follow-up. Significant aneurysm sac shrinkage (> or = 5 mm) occurred in 73.1 % (19/26) vs. 43.1% (28/65) of patients in the Zenith and AneuRx groups, respectively, at 12 months (p = 0.03). At 6 months follow-up, sac shrinkage rates were 51.4% (19/37) vs. 25.8% (16/62) in the Zenith and AneuRx groups, respectively (p = 0.04). Mean reduction of sac diameter at 12 months was -7.6 +/- 1.6 mm vs. -3.5 +/- 0.8 mm in the Zenith and AneuRx groups, respectively (p = 0.01). There was a trend toward fewer Type I and III endoleaks at 1 month in the Zenith group (0 vs. 8.3%) that did not achieve statistical significance (p = 0.067). The presence of any endoleak (> or = 1 month) was associated with reduced 12 month shrink rates from 47.1% (25/51) to 28% (4/14) in the AneuRx group (p = 0.35) and from 77.3% (17/22) to 50% (2/4) in the Zenith group (p = 0.25). Patients treated with the Zenith endograft demonstrated a significantly higher rate and amount of aneurysm sac shrinkage than patients treated with an AneuRx device. Endoleaks appeared to negatively influence shrink rates with both endografts. PMID- 12545253 TI - Endovascular treatment of failed prior abdominal aortic aneurysm repair. AB - Failure of endovascular or conventional abdominal aortic aneurysm (AAA) repair may occur as a result of attachment site endoleak (type I) or paraanastomotic aneurysm and pseudoaneurysm formation. This study examined the results of the use of secondary endovascular grafts for the treatment of failed prior infrarenal AAA repair procedures. Forty-seven patients were treated with endovascular grafts. These included 14 patients with type I endoleaks (5 proximal, 8 distal, 1 proximal and distal) and 33 patients with paraanastomotic aneurysms after standard open surgical AAA repair (3 proximal aorta, 5 distal aorta, 21 iliac, 4 proximal and distal). The interval between the primary aortic procedure and the endovascular repair was significantly shorter for failed endovascular procedures (mean, 18.2 months; range, 1-42 months) than for failed conventional procedures (mean, 108.9 months; range, 12-216 months) (p <0.01). The endovascular devices used for correction of the failed AAA repairs were Talent (23), physician-made (19), AneuRx (2), Vanguard (2), and Excluder (1). Transrenal fixation was used for repair of all proximal anastomotic failures. Mean follow-up after reintervention was 12.2 months in patients with failed endovascular grafts and 10.6 months in patients with failed conventional grafts. Patient demographics were as follows: average age, 78 years; 36 male and 11 female; and 4.1 comorbid medical conditions per patient. The endovascular graft was successfully deployed in all 47 cases; 1 patient experienced a persistent proximal attachment site endoleak after endograft deployment. Endovascular grafts may be used to treat previously failed endovascular and conventional AAA repair procedures with good short- and intermediate-term results. Endovascular treatments in these cases may avoid the difficulties of aortic reoperation or AAA repair in the setting of prior endovascular aortic grafting. PMID- 12545254 TI - Thoracodorsal sympathectomy for severe hyperhydrosis: posterior bilateral versus unilateral staged sympathectomy. AB - The aim of this study was to compare the results of simultaneous bilateral thoracodorsal sympathectomy in the prone position with those of anterolateral sympathectomy performed in two staged, separate procedures for the treatment of bilateral excessive sweating of the hands and axillae, and to describe our technique for bilateral, simultaneous thoracodorsal sympathectomy. From July 1995 to March 2001, 202 thoracodorsal sympathectomies were done in 101 patients for severe hyperhydrosis. There were 79 females (age range 20-46) and 22 males (age range 19-65). In 52 patients, anterolateral sympathectomies were performed in the supine position, using unilateral lung collapse, with both sides operated on in two separate, staged procedures. In 49 patients, bilateral sympathectomy was conducted during a single procedure, in the prone position, without using unilateral lung collapse. In comparing the results from these two methods, we concluded that simultaneous bilateral thoracodorsal posterior sympathectomy, has comparable safety, may improve outcome, decreases in half the number of hospital admissions, and produces a significant overall reduction in cost when compared with staged anterolateral sympathectomy for the treatment of severe hyperhydrosis. PMID- 12545255 TI - Imaging of abdominal aortic aneurysm: current status. PMID- 12545256 TI - Debulking or biopsy of malignant glioma in elderly people - a randomised study. AB - BACKGROUND: Patients with radiologically (MRI and/or CT images) suspected malignant glioma is referred to radiotherapy after craniotomy and resection of the tumour or after diagnostic biopsy. Patients with poor preoperative status and elderly patients are diagnosed more often by biopsy and treated by radiotherapy rather than by craniotomy and tumour resection. However, based on previous retrospective studies it is not possible to conclude which procedure is better for elderly patients. Thus a prospective study comparing these two procedures with elderly patients was planned. METHODS: 30 patients older than 65 years with radiologically (CT and/or MRI) obvious malignant glioma were randomised into two groups: I) stereotactic biopsy and II) open craniotomy and resection of the tumour. Nineteen patients were diagnosed to have grade IV glioma and four patients grade III glioma. Seven out of 30 (23%) were followed in the "intention to-treat" group with diagnosis of stroke (n=3), metastasis (n=2), malignant lymphoma (n=1) and one with out histological diagnosis. Patients with histologically verified malignant glioma (grade III-IV) were diagnosed by stereotactic biopsy (n=13) or by open craniotomy and resection (n=10) and all the patients were referred to radiotherapy. Survival and time of deterioration were followed. FINDINGS: The overall median survival time was 146 (95% CI 89-175) days after the procedure. The estimated median survival time was 171 (95% CI 146-278) days after the craniotomy versus 85 (95% CI 55-157) days after the biopsy (p=0.035). The estimated survival time was 2.757 times longer (95% CI 1.004 7.568, p=0.049) after craniotomy. However, there was no significant difference in the time of deterioration between these two treatments (p=0.057). Amount of radiotherapy given had a significant effect on survival (p=0.001). INTERPRETATION: Longer survival time is achieved after open craniotomy and resection of tumour. However, overall benefit of open surgery to patient seems to be modest, while time of deterioration did not differ between two treatment groups. Our results support previous studies on the benefit of radiotherapy in the treatment of malignant glioma. PMID- 12545257 TI - Prospective comparative study of the effectiveness of epidural morphine and ropivacaine for management of pain after spinal operations. AB - OBJECTIVE: Evaluation of the effectiveness of local application of morphine or ropivacaine for treatment of local and radicular pain after lumbar disc operations. Critical review of the literature about the possibilities of management of postoperative pain after spinal operations. METHODS: A total of 113 patients were randomly given 5 mg morphine sulfate (N=42), 10 ml 0,25% ropivacaine (N=42) or physiological NaCl solution (N=21) locally after lumbar disc operation before wound closure. Postoperative lumbar and radicular pain was scored by the patients from 0 to 10 and registered on the evening of the day of operation and on the 1., 2., 3., and 5. days. Mean pain numbers of the 3 groups have been compared, subdivided into local lumbar and in radicular pain. Our own results have been compared with the results of reports in the literature. RESULTS: In our own study the morphine group had less lumbar and less radicular pain on all 5 days than both of the two other groups. This difference was statistically significant on days 0, 1, 2, and 3. The ropivacaine group was on all days less effective than the morphine group, better than the placebo group on the operation and first day, but the difference against the placebo group was statistically not significant. CONCLUSION: Local application of 5 mg morphine sulfate is effective in prevention or reduction of postoperative lumbar and radicular pain after lumbar disc operations. Ropivacain is less effective. The routine application of epidural morphine at the end of spinal operations can be recommended. It also can be justified to try to prolong the morphine effect by mixing it into a paste as described by Needham and by Hurlbert, and to irrigate the operative field with ropivacaine at the end of the operation. PMID- 12545258 TI - Transvenous embolisation of dural carotid-cavernous fistulas by multiple venous routes: a series of 27 cases. AB - BACKGROUND: Transvenous embolisation is an effective option in the treatment of dural carotid-cavernous fistulas (DCCFs). This can be achieved via a number of venous routes. METHOD: From 1997 to 2001, 27 patients with DCCFs (15 unilateral and 12 bilateral) were treated by transvenous embolisation with Guglielmi detachable coils and fibered platinum coils. The transvenous embolisation routes included inferior petrosal sinus (IPS) alone (18 patients), IPS and inter cavernous sinus (6 patients), IPS and clival plexus (1 patient), superior ophthalmic vein (SOV) via facial vein (1 patient) and SOV via superficial temporal vein (1 patient). One patient required further transarterial embolisation with polyvinyl alcohol particles. FINDINGS: The follow-up period ranged from 4 to 57 months (average 26 months). Two patients had transient ophthalmoplegia and 2 patients had symptomatic recurrence of the DCCF during the follow-up. There was no permanent procedure-related morbidity. Clinical cure was achieved in 26 patients (96%) and complete angiographic obliteration was documented in 24 patients (89%). INTERPRETATION: DCCFs can be successfully treated by transvenous embolisation via different venous routes. PMID- 12545259 TI - Management of benign craniovertebral junction tumors. AB - BACKGROUND: We report our surgical experience in the treatment of fifteen consecutive patients with benign craniovertebral junction tumors, observed from 1993 to 2000 at our department. METHOD: We treated 7 meningiomas, 3 epidermoids, 3 C1 neurinomas and 2 neurinomas of the lower cranial nerves. Clinical results were evaluated by Karnofsky Performance Scale and all patients underwent preoperative neuroradiological evaluation with CT, MRI and MRA; angiography was not routinely performed and was considered for each individual case. FINDINGS: 11 partial transcondilar and 4 retrocondilar approaches were performed. Total removal was achieved in 11 cases (73,3%) and subtotal removal in 4 patients (26,7%). None of the patients required occipitocervical fusion. Patients were followed for an average period of 24+/-31 months. Clinical and radiological follow-up showed no recurrence in cases with total removal. In all patients a statistically significant postoperative increase of KPS scores was recorded. The treatment of epidermoid tumors presented particular issues: debulking the lesion, we obtained a surgical window, avoiding a large removal of bone. In Nakasu grade 1 or 2 meningiomas, we carried out total removal by piecemeal resection and without complete condylectomy and bone fixation. INTERPRETATION: The choice of these approaches and the extent of bone resection should be defined according to the tumor's location and size. Moreover we emphasize that preoperative neuroradiological evaluations on presumptive tumor type could be helpful to the surgeon in order to tailor the technique to different lesions, providing the required exposure, without unnecessary surgical steps. PMID- 12545260 TI - Serum- and CSF-concentrations of brain specific proteins in hydrocephalus. AB - OBJECT: Hydrocephalus is characterised by elevated intracranial pressure (ICP) and gives rise to brain damage. The aim of this study was to investigate the significance of brain specific proteins as markers in the evaluation of brain damage in hydrocephalus. Therefore we determined the levels of four brain specific proteins in cerebrospinal fluid (CSF) and serum of symptomatic hydrocephalic patients. METHODS: During 41 CSF shunt-operations (both primarily placed shunts and shunt-revisions) CSF and blood samples were obtained and analysed for neuron-specific enolase (NSE), S-100b, glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP). The results were compared with an age-matched control group. Patients with varying clinical symptoms, denoting different levels of increased intracranial pressure prior to surgery, were included in this study. RESULTS: We observed significantly increased CSF-levels of S-100b and GFAP in the hydrocephalic patients, whereas NSE and MBP were markedly increased only in patients with very severe symptoms. Serum levels of all proteins were only minimally increased and did not correlate with CSF-levels. The slightly elevated levels of CSF-NSE in most of the patients suggest only subtle neuronal damage, which is not related to permanent neurological symptoms. The elevated levels of S-100b and GFAP are indicative of a reactive astrogliosis, which has also been demonstrated in histopathological studies. No demyelination seems to occur, according to the normal levels of MBP observed in this study. CONCLUSIONS: Although CSF levels of brain specific proteins are elevated in hydrocephalic patients, indicating brain damage due to hydrocephalus, neither CSF nor serum-concentrations of brain specific proteins seem to be valuable tools in the clinical evaluation of the severity of hydrocephalus. PMID- 12545261 TI - Publication bias in neurosurgery: lessons from series of unruptured aneurysms. AB - BACKGROUND: Bias favouring publication of research with "positive" results over studies with "negative" results is widely suspected. The present investigation addressed this problem in the field of neurosurgery through a review of recent literature concerning outcome of surgery for unruptured intracranial aneurysms. METHODS AND FINDINGS: A Medline search was performed seeking case series of surgical treatment for unruptured intracranial aneurysms that analyzed 50 or more patients. Ten type I studies (retrospective studies from a single institution; 1457 patients) met these entry criteria. In general, type I studies reported excellent surgical outcome, with mean combined mortality and morbidity of 7.8% (95% confidence interval (CI), 6.4% to 9.2%). We found 4 multicenter or community based studies (type II studies; 5401 patients). Mean combined mortality and morbidity in the type II studies was 20.3% (95% CI, 19.2% to 21.4%), much higher than in type I studies. Relative risk was 2.6 (95% CI, 2.2 to 3.1) for patients in type II studies compared with those in type I studies. Two meta-analyses (type III studies) described combined mortality and morbidity of 5.0% and 12.7%, essentially corresponding to results of type I studies. INTERPRETATION: Publication bias is present in the neurosurgical literature. Studies with an excellent surgical outcome are more likely to be published than those with an average outcome. Conclusions of literature reviews or meta-analyses based on published work therefore may be misleading. The solution to the problem would be community-based prospective registration of all patients who underwent surgery, providing a sampling frame free from publication bias. PMID- 12545262 TI - Effects of intraventricular infusion of vascular endothelial growth factor on cerebral blood flow, edema, and infarct volume. AB - BACKGROUND: Therapeutic cerebral angiogenesis, utilizing angiogenic factors to enhance collateral vessel formation within the central nervous system, is a potential method for cerebral revascularization. A prior dose-response study determined that intracerebroventricular infusion of vascular endothelial growth factor (VEGF) increases vascular density with minimal associated brain edema at a concentration of 5 microg/ml. The purpose of this study was to assess effects of intracerebroventricular infusion of VEGF (5 microg/ml) on cerebral blood flow, infarct volume, and brain edema after ischemia. METHODS: Recombinant human VEGF(165) was infused into the right lateral ventricle of rats with an osmotic minipump at a rate of 1 microl/hr for 7 days. Control animals received vehicle only. Ischemia was produced by transient (2 hours) middle cerebral artery occlusion (MCAO). After MCAO, cerebral blood flow was determined with the indicator fractionation technique: infarct volume was assessed with 2,3,5 triphenlytetrazolium chloride staining, and brain edema was determined by measuring brain water content. FINDINGS: Cerebral blood flow was not significantly different in animals treated with VEGF compared to controls. There was a significant reduction in total infarct volume after temporary MCAO in VEGF treated animals compared to controls (163+/-37 mm(3) vs. 309+/-54 mm(3), P<0.05). Brain water content after transient MCAO was also significantly reduced in VEGF treated animals compared to controls (80.9+/-0.7% vs. 83.3+/-0.6%, P<0.05). INTERPRETATION: Intracerebroventricular infusion of VEGF(165) (5 microg/ml) decreases infarct volume and brain edema after temporary MCAO without a significant increase in cerebral blood flow. These results indicate that VEGF may have a direct neuroprotective effect in cerebral ischemia. PMID- 12545263 TI - Repositioning of the vertebral artery with titanium bone fixation plate for trigeminal neuralgia. AB - BACKGROUND: Trigeminal neuralgia is usually treated by the padding method using Teflon felt. However this can not be done in certain cases in whom a large tortuous vertebrobasilar artery compresses the fifth nerve. The transposition method using the sling may be an alternative method. But this method is not an easy procedure and requires a relatively large craniotomy. Two cases were treated by a new and simpler effective technique. CLINICAL PRESENTATION: Two cases of the trigeminal neruralgia were treated. The first case was a 71 year-old male and the second case was a 63 year-old male. The history of the medical treatments were similar and both cases had had trigeminal nerve blocks and were prescribed carbamazepin. However, the pain control was insufficient in both cases. In both cases, three dimensional computerized tomography showed the large tortuous right vertebral artery ran just behind the clivus and compressed the right trigeminal nerve. In the second case past history showed a recent hypertensive cerebellar hemorrhage. TECHNIQUE AND RESULTS: A right suboccipital craniotomy were performed in both cases. In both cases, the right vertebral artery compressed the trigeminal nerve in a rostral direction. The sling technique with nylon sutures was tried in both cases but failed during surgery. Then, the bone fixation stainless plate was cut to 10 cm in length and pre-shaped with pliers. After being shaped, the distal end of the plate was inserted between the vertebral artery and fifth nerve and the proximal end of the plate was fixed to the skull by screw. The fifth nerve was completely isolated from the artery as they were in direct contact. After surgery, the pain disappeared completely during the follow up of one and a half year in the first case and 9 months in the second case. CONCLUSION: The plate can be bent and curved with plier to suit each individual case. This technique is easily applied even when the slings or other isolation technique is not available and appeared to achieve the mechanically stronger reposition and fixation of a very large and tortuous artery away from the trigeminal nerve. PMID- 12545264 TI - Intraoperative direct neuroendoscopic observation of the aqueduct in Dandy-Walker malformation. AB - A 3-month-old female infant with Dandy-Walker malformation manifesting as hydrocephalus was treated successfully by only ventriculoperitoneal shunting. A flexible neuroendoscope was used intraoperatively to confirm the patency of the aqueduct, i.e. communication of the ventricular system and the cyst in the posterior fossa. Direct confirmation of the patency of the aqueduct and cyst communication is valuable to select the shunt procedure in the treatment of Dandy Walker malformation. PMID- 12545265 TI - Infection-related atlantoaxial subluxation in two adults: Grisel syndrome or not? AB - Grisel's syndrome involves the subluxation of the atlanto-axial joint from inflammatory ligamantous laxity following an infectious process in the head or neck. It is a rare disease usually affecting children, but infrequent adult cases do occur. Today, due to the widened use of antibiotics and availability of MR imaging, some cases presenting with neck pain, resulting from the infectious inflammation of C1 and C2 but without pronounced subluxation, can be a challenging problem for most neurosurgeons. Several theories have been proposed to explain the pathogenesis of inflammatory subluxation. The primary treatment of Grisel's syndrome is medical. The underlying infectious organism must be isolated and appropriate antibiotics must be prescribed. The subluxation is reduced in holter or skeletal traction. This paper reports two cases of infection-related atlanto-axial subluxation in two adults. The literature on this subject is briefly reviewed. PMID- 12545266 TI - Non-obstructive hydrocephalus associated with intracranial schwannomas: hyperproteinorrhachia as an etiopathological factor? AB - BACKGROUND: This series illustrates the association of communicating hydrocephalus with intracranial non-obstructive schwannomas. This association has commonly been observed, however it has only been reported once previously. Moreover, in all the patients we present, hyperproteinorrhachia was a common denominator. This finding may therefore be the underlying mechanism for hydrocephalus. METHOD AND FINDINGS: Seven patients presenting with intracranial schwannomas along with non-obstructive hydrocephalus and hyperproteinorrhachia are reported. Six had a vestibular schwannoma and presented with a unilateral deafness and various degrees of gait disturbance, urinary incontinence and neuropsychological impairment. Due to their advanced ages, these patients underwent a ventriculo-peritoneal shunt, and their symptoms related to hydrocephalus resolved. One patient that suffered from hemifacial dysesthesia and memory deficits presented with a non-obstructive trigeminal schwannoma. In this case the tumour was removed and the hydrocephalus was consequently reversed, and the CSF protein content normalized. INTERPRETATION: The constant finding of hyperproteinorrhachia in all these patients suggests that a high CSF protein content may be the underlying cause of hydrocephalus through a speculative mechanism of decreased CSF resorption. PMID- 12545267 TI - Late hydrocephalus in a case of wandering bullet into the pineal region. AB - We report a patient in whom a bullet in the brain migrated into the pineal region causing hydrocephalus 3 months later. In patients undergoing surgical removal of intracerebral or intraventricular bullets, it is recommended to obtain an x-ray or CT scan on the day of operation. Late hydrocephalus may occur several months after migration of the bullet due to scar tissue. PMID- 12545268 TI - Orbital syndrome after a surgery of vestibular schwannoma via middle fossa approach. PMID- 12545270 TI - Intrahepatic lithiasis as a late complication of hepaticojejunostomy. AB - The development of intrahepatic lithiasis proximal to a bilodigestive anastomosis is rare. We report a case of intrahepatic lithiasis of the right hepatic duct, which developed almost 6 years after a hepaticojejunostomy performed for an iatrogenic lesion of the common hepatic duct. The patient experienced repeated episodes of acute cholangitis and was treated with intracorporeal lithotripsy and dilation of the stenosis. PMID- 12545271 TI - Mediastinalemphysema after laparoscopic gastric banding. AB - From January 1996 to July 2001, 530 patients underwent a laparoscopic gastric banding operation at the University Hospital of Innsbruck. In one of these patients, a pneumomediastinum developed 17 months after surgery. Unable to swallow, the 28-year-old patient suddenly felt sick and reported left chest pain. Additionally, recurrent vomiting and later hematemesis were reported. A band migration was excluded via endoscopy. After the diagnosis of a mediastinalemphysema was established, the patient was treated conservatively. The causes of mediastinalemphysema are heterogeneous and sometimes not exactly clear. In our patient it is a question whether vomiting led to an esophageal tear, which in turn was responsible for the mediastinalemphysema. PMID- 12545273 TI - Role of laparoscopy in blunt perforations of the small bowel. AB - BACKGROUND: The purpose of this study was to determine whether laparoscopy could improve our ability to diagnose and treat perforations of the small bowel. METHODS: From 1985 to 2001, among 250 patients admitted for a blunt abdominal trauma, 195 surgical explorations were performed, comprising 42 laparoscopies. On admission, 108 patients underwent an abdominal ultrasonography (US) and 104 a computed tomography (CT). Thirty-nine patients had a blunt small bowel trauma (BSBT) that was explored and/or treated by laparoscopy (n = 15) or celiotomy (n = 24). We compared the patients who underwent emergency surgery (23) and those who were operated on after a mean delay of 51 h. RESULTS: On admission, for detecting free intraperitoneal fluid with a BSBT, the sensitivity of CT was 93.3% and specificity was 13.6%. Regarding the 23 patients who underwent emergency surgery, 8/20 had positive US and 10/15 had positive CT, whereas the 15 celiotomies and 8 laparoscopies indicated BSBT. Regarding the 16 patients who underwent delayed surgery, 9 patients underwent a celiotomy and 7 a laparoscopy; all BSBTs were diagnosed. In searching for BSBT, the sensitivity and specificity of laparoscopy were both 100%, whereas the sensitivity of CT was 83.3% and specificity was 22.2%. Twenty of 23 patients undergoing emergency surgery had bowel suture repair compared to 3/16 after delayed surgery. In contrast, a resection was necessary in 10/12 patients treated after a delay compared to 2/17 operated in emergency. Five BSBTs were sutured by laparoscopy, whereas 10 required conversion. Laparoscopy avoided celiotomy in 15 cases. There were no deaths and no enterostomy. CONCLUSION: In hemodynamically stable patients with blunt abdominal trauma, laparoscopy safely and effectively identifies small bowel injuries. Early recognition of these injuries and timely surgical treatment offer the best prognosis. PMID- 12545272 TI - Emergency department visits by demented patients with malfunctioning feeding tubes. AB - BACKGROUND: Objective data indicate that feeding tubes in demented patients may not be efficacious and can have serious complications, but no study characterizes emergency department resource utilization for these patients. This study aimed to evaluate the incidence and resource utilization related to feeding tube malfunction in demented patients visiting the emergency department. METHODS: A retrospective chart review for all demented patients visiting the emergency department with malfunctioning feeding tubes from September 1999 to May 2001 was conducted. Demographic data, diagnoses, type of tube, length of emergency department stay, method of transportation to the emergency department, consultations, laboratory evaluation, x-ray data, and total hospital and ambulance charges were determined. RESULTS: A total of 138 emergency department visits by 33 patients occurred during this period (range of visits per patients, 1-21; mean, 4.1 +/- 4.3). Malfunctions occurred in 61 percutaneous endoscopically placed gastric tubes, 37 jejunostomy tubes, 34 gastric tubes, 4 endoscopically placed gastrostomy and jejunostomy tubes, and 2 percutaneous endoscopically placed jejunostomy tubes. This required 108 ambulance round-trips to and from the emergency department. The most frequent complication was unintentional dislodgement (n = 125). The average length of stay was 2.6 +/- 1.6 h. All the patients were seen by an emergency department physician. In addition, there were 99 surgical and 26 gastroenterology consultations about these patients. The total hospital charges, not including physician fees, were 86,234.48 dollars, and the total reimbursement (actual) from Medicare for ambulance charges was 57,664.00 dollars. During the same 21-month period, 42 feeding tubes were placed for dementia. CONCLUSIONS: The expense of emergency department visits for tube dislodgment or malfunction is a previously unreported issue involved in the tube feeding of demented patients. Extrapolation of our data yields an estimated health care charge of almost $11 million for the country per year. PMID- 12545274 TI - Relation of risk of systemic lupus erythematosus to west African admixture in a Caribbean population. AB - Risk of systemic lupus erythematosus (SLE) is higher in people of west African descent than in Europeans. The objective of this study was to distinguish between genetic and environmental explanations for this ethnic difference by examining the relationship of disease risk to individual admixture (defined as the proportion of the genome that is of west African ancestry); 124 cases of SLE and 219 matched controls resident in Trinidad were studied. Analysis of admixture was restricted to 52 cases and 107 controls who reported no Indian or Chinese ancestry. These individuals were typed with a panel of 26 single-nucleotide polymorphisms and five insertion/deletion polymorphisms chosen to have large allele frequency differentials between west African, European and Native American populations. A Bayesian model for population admixture, individual admixture and locus ancestry was fitted by Markov chain simulation. Mean west African admixture (M) was 0.81 in cases and 0.74 in controls (P=0.01). The risk ratio for SLE associated with unit change in M was estimated as 32.5 with a 95% confidence interval (CI) of 2.0-518. Adjustment for measures of socioeconomic status (household amenities in childhood and years of education) altered this risk ratio only slightly (adjusted risk ratio: 28.4, 95% CI 1.7-485). These results support an additive genetic model for the ethnic difference in risk of SLE between west Africans and Europeans, rather than an environmental explanation or an "overdominant" model in which risk is higher in heterozygous than in homozygous individuals. This conclusion lays a basis for localizing the genes underlying this ethnic difference in risk of SLE by admixture mapping. PMID- 12545276 TI - Duplication of the MID1 first exon in a patient with Opitz G/BBB syndrome. AB - Opitz G/BBB syndrome is a malformation syndrome of the ventral midline mainly characterized by hypertelorism, swallowing difficulties, hypospadias and developmental delay. SSCP analysis and genomic sequencing of the MID1 open reading frame have identified mutations in 80% of the families with X-linked inheritance. However, in many patients the underlying genetic defect remains undetected by these techniques. Using RNA diagnostics we have now identified a duplication of the MID1 first exon in a patient with X-linked Opitz G/BBB syndrome. This duplication introduces a premature termination codon. In addition, we could significantly lower the threshold for mutation detection on the DNA level by combining SSCP analysis with DHPLC technology. PMID- 12545278 TI - A rotation and translation invariant discrete saliency network. AB - We describe a neural network that enhances and completes salient closed contours in images. Our work is different from all previous work in three important ways. First, like the input provided to primary visual cortex (V1) by the lateral geniculate nucleus (LGN), the input to our computation is isotropic. That is, it is composed of spots, not edges. Second, our network computes a well-defined function of the input based on a distribution of closed contours characterized by a random process. Third, even though our computation is implemented in a discrete network, its output is invariant to continuous rotations and translations of the input image. PMID- 12545279 TI - Traveling-wave pattern generator controls movement and organization of sensory feedback in a spinal cord model. AB - A traveling wave in a two-dimensional spinal cord model constitutes a stable pattern generator for quadruped gaits. In the context of the somatotopic organization of the spinal cord, this pattern generator is sufficient to generate stable locomotive limb trajectories. The elastic properties of muscles alone, providing linear negative feedback, are sufficient to stabilize stance and locomotion in the presence of perturbative forces. We further show that such a pattern generator is capable of organizing sensory processing in the spinal cord. A single-layer perceptron was trained to associate the sensory feedback from the limb (coding force, length, and change of length for each muscle) with the two dimensional activity profile of the traveling wave. This resulted in a well defined spatial organization of the connections within the spinal network along a rostrocaudal axis. The spinal network driven by peripheral afferents alone supported autonomous locomotion in the positive feedback mode, whereas in the negative feedback mode stance was stabilized in response to perturbations. Systematic variation of a parameter representing the effect of gamma-motor neurons on muscle spindle activity in our model led to a corresponding shift of limb position during stance and locomotion, resulting in a systematic displacement alteration of foot positions. PMID- 12545275 TI - Human genetic disease caused by de novo mitochondrial-nuclear DNA transfer. AB - Transfer of nucleic acid from cytoplasmic organelles to the nuclear genome is a well-established mechanism of evolutionary change in eukaryotes. Such transfers have occurred throughout evolution, but so far, none has been shown unequivocally to occur de novo to cause a heritable human disease. We have characterized a patient with a de novo nucleic acid transfer from the mitochondrial to the nuclear genome, a transfer that is responsible for a sporadic case of Pallister Hall syndrome, a condition usually inherited in an autosomal dominant fashion. This mutation, a 72-bp insertion into exon 14 of the GLI3 gene, creates a premature stop codon and predicts a truncated protein product. Both the mechanism and the cause of the mitochondrial-nuclear transfer are unknown. Although the conception of this patient was temporally and geographically associated with high level radioactive contamination following the Chernobyl accident, this case cannot, on its own, be used to establish a causal relationship between radiation exposure and this rare type of mutation. Thus, for the time being, it must be considered as an intriguing coincidence. Nevertheless, these data serve to demonstrate that de novo mitochondrial-nuclear transfer of nucleic acid is a novel mechanism of human inherited disease. PMID- 12545280 TI - Emergence of coherent traveling waves controlling quadruped gaits in a two dimensional spinal cord model. AB - The concerted and self-organizing behavior of spinal cord segments in generating locomotor patterns is modulated by afferent sensory information and controlled by descending pathways from the brainstem, cerebellum, or cortex. The purpose of this study was to define a minimal set of parameters that could control a similar self-organizing behavior in a two-dimensional neural network. When we implemented synaptic depression and active membrane repolarization as two properties of the neurons, the two-dimensional neural network generated traveling waves. Their wavelength and angle of propagation could be independently controlled by two parameters that modulated excitatory premotor neurons and inhibitory commissural neurons. It is further demonstrated that the selection of wave parameters corresponds to the selection of quadruped gaits. PMID- 12545281 TI - Computational model of the cockroach escape behavior: winner and losers in a population code. AB - I present a comprehensive biologically oriented computational model to account for the escape response of the cockroach on the ground. This model is an expansion of previous work that accounted only for discriminating left from right wind directions [Ezrachi et al. (1999) Biol Cybern 81: 89-99]. The model is composed of computational elements describing the biological processes taking place in the various neurons and includes input which emulates empirical data. With this model it is possible to obtain escape behavior that resembles natural behavior. The model is used to address an ongoing debate as to whether the cockroach's turn direction is determined by computations carried out by the entire neuronal population (PC) or rather by a "winner-take-all" (WTA) mechanism. I suggest that the computation mechanism that underlies the cockroach escape response is composed of both PC and WTA principles. Based on the properties of the suggested new mechanism I denote it a "Darwinian population code." PMID- 12545282 TI - An alternative approach to synthesizing bipedal walking. AB - Based on mechanical analysis, three gait descriptors are found which should be controlled to generate cyclic gait of a seven-link humanoid biped in the sagittal plane: (i) step length, (ii) step time, and (iii) the velocity of the center of mass (CoM) at push off. Two of these three gait descriptors can be chosen independently, since the CoM moves almost ballistically during the swing phase. These gait descriptors are formulated as end-point conditions and are regulated by a model predictive controller. In addition, continuous controls at the trunk and knees are implemented to maintain the trunk upright and to ensure weight bearing. The model predictive controller is realized by quadratic dynamic matrix control, which offers the possibility of including constraints that are exposed by the environment and the biped itself. Specifying step length and CoM velocity at push off, the controller generates a symmetric and stable gait. The proposed control scheme serves as a general-purpose solution for the generation of a bipedal gait. The proposed model contains fewer parameters than other models, and they are all directly related to determinants of bipedal gait: step length, trunk orientation, step time, walking velocity, and weight bearing. The proposed control objectives and the model of humanoid bipedal walking have potential applications in robotics and rehabilitation engineering. PMID- 12545283 TI - Evidence for a specific internal representation of motion-force relationships during object manipulation. AB - Human subjects learned a tracking task which required them to point at a moving target with the free end of an inverted pendulum object. In order to determine how subjects represented this object internally, we studied learning interference between variants of this task in which the pendulum object had either stable or unstable dynamics. Using a novel method, agreement between possible internal representations of the two tasks was estimated by analysis of the motion-to torque relationships experienced by each subject as they manipulated each object. It was possible to predict retention of the primary task on day 2 from our measure of agreement between primary and interfering tasks on day 1. This result suggests that the subjects learned the correct torque patterns to use to produce specific desired patterns of motion as they learned the balancing task. Surprisingly, the analyses indicate that retention was not impaired when similar motions of the two objects required retrieval of incompatible torque responses, but retention was impaired when similar patterns of motion in the two tasks required similar patterns of applied torque. These findings can be accounted for by a simple model of how multiple similar torque responses are selected and retrieved from memory when responses are freely chosen. PMID- 12545284 TI - Time versus size: which characteristic of a neural response carries more information? AB - Considering a variety of quite different candidate neural codes, Cheng and Wasserman [(1996) Biol Cybern 75, 93-103, 105-115] reported data which suggested that amplitude codes transmitted information more faithfully than temporal codes. Reanalyzing their data, the present study measured how well size and time represented information resident within the same neural response feature, namely the response peak. Responses from photoreceptor cells and optic nerve fibers in the peripheral visual system of Limulus polyphemus were therefore re-examined using signal detectability values provided by receiver-operating-characteristic analyses to compare how well the timing and the amplitude of the response peak represent information within a single cell and how well they transmit information between cells. Data were available for several cells in several different light adaptation states onto which several different test-flash intensities had been superimposed. The present reanalysis of these data replicated the results yielded earlier by the peak candidate code, and compared them with the data produced by two different measurements of the timing of the same response peak. A relative timing code was derived from measurements of the time that elapsed between the moment when the response exceeded a criterion potential and when it reached its peak. An absolute-timing code was derived from the time that elapsed between test flash onset and the peak. The results clearly indicated that the peak code represented information within cells better than either of the two timing codes. However, both peak and absolute-timing codes clearly transmitted the available information between cells more faithfully than the relative-timing code. These data lead to two conclusions. First, that the same response feature, when measured in different ways, can produce remarkably different information processing outcomes. In this particular case, time represented information resident within a cell less well than did size. Second, that the fidelity of information transmission between nerve cells may be relatively independent of the quantity of information resident within any particular cell. PMID- 12545285 TI - Sectorial keratitis and uveitis: differential diagnosis. AB - PURPOSE: The purpose of this study was to examine the importance of considering the differential diagnosis for patients with sectorial keratitis and uveitis by case summary and literature review. METHODS: A retrospective review of patients with sectorial keratitis and uveitis seen at the Ocular Immunology and Uveitis Service of the Massachusetts Eye and Ear Infirmary and a summary of the diagnoses of cases with similar ocular findings that have been reported in the literature. RESULTS: Data on six patients with sectorial keratitis and uveitis were reviewed. Four patients were female and two were male, ages 21-50 years. All were eventually diagnosed with herpes simplex viral stromal sectorial keratitis. The corneal infiltrates were most common in the superior corneal quadrants, located in the posterior corneal layers. Anterior non-granulomatous uveitis was present in all cases. One case had bilateral ocular involvement. Five of the six patients responded to topical steroids and antiviral treatment. Immunomodulation with methotrexate, cyclosporine, and systemic prednisone was required in one patient. Glaucoma was a complication in six of the seven eyes. Five patients developed corneal scarring, localized in the anterior and mid-stroma. CONCLUSION: The differential diagnosis of sectorial keratitis and uveitis is limited. The entities included in the differential are diverse; some of them threaten not only vision but also life. It is important for the ophthalmologist to be familiar with these entities, in order to pursue pertinent diagnostic investigations and arrive at an accurate diagnosis and institute appropriate management. PMID- 12545286 TI - Detection of visual activation of lateral geniculate nucleus by positron emission tomography. AB - PURPOSE: Although activation of the lateral geniculate nucleus (LGN) has been shown by functional magnetic resonance imaging, a quantitative evaluation of the activity has not been reported by positron emission tomography (PET). The purpose of this study was to quantify the degree of activation of the LGN and other vision-related structures by visual stimuli as detected by PET. METHODS: PET activation was induced by a bolus injection of 15O-labelled water (H2(15)O) in six normal volunteers. The regional cerebral blood flow (rCBF) was measured under three conditions: eyes closed, binocular flickering light stimulation, and binocular viewing of a video movie. Single-subject analysis, group analysis, and correlation analysis were performed. RESULTS: Although single-subject analysis did not show significant changes, group analysis revealed a significant increase in rCBF in the left LGN induced by flickering light stimulation ( P<0.01) and video stimulation ( P<0.05). Localization of the activation foci in the LGN agreed with the retinotopic organization of the LGN. CONCLUSION: Although the LGN is relatively small compared with the resolution of the PET image, visually activated blood flow changes could be determined by H2(15)O PET: PMID- 12545287 TI - Retinal hemorrhage after photodynamic therapy in patients with subfoveal choroidal neovascularization caused by age-related macular degeneration. AB - PURPOSE: To report the frequency and the evolution of the extensive retinal hemorrhages that can appear within 48 h after the application of photodynamic therapy. METHODS: Two hundred and fifteen individual eyes of 194 consecutive patients with subfoveal choroidal neovascularization (CNV) due to age-related macular degeneration who underwent photodynamic treatment were included in the study. The visual acuity was measured before and after treatment. Color and red free photographs were taken. Fluorescein angiography and optical coherence tomography (OCT) were also performed in order to describe the macular hemorrhages. RESULTS: Four out of 215 eyes developed macular hemorrhage within 48 h after the photodynamic therapy. Before treatment one of the four patients had classic CNV, one predominantly classic and two patients occult CNV without any classic component. In all four cases, the hemorrhage after photodynamic therapy (PDT) was extensive, it extended beyond the arcades and it was not absorbed during the follow-up period, which ranged from 11 to 21 months. The greatest linear dimension of the hemorrhage was extremely high (>12,000 microm). CONCLUSION: Extensive macular hemorrhage was observed in 1.86% of the studied cases. The hemorrhage was not related to the type of the CNV lesion before treatment. The size and the appearance of hemorrhage within 48 h after treatment support the view that the hemorrhage is a direct consequence of the photodynamic therapy and not related to the natural course of the disease. PMID- 12545288 TI - The negative correlation between age and intraocular pressures measured nyctohemerally in elderly normal-tension glaucoma patients. AB - PURPOSE: To evaluate the correlation between age and intraocular pressure (IOP) measured every 3 h for 24 h in older normal-tension glaucoma (NTG) patients. METHODS: Ninety consecutive patients suspected of having NTG on the basis of both visual field defect and disc appearance were hospitalized for measurement of nyctohemeral IOP every 3 h over a 24-h period with a Goldmann tonometer after a 4 week wash-out of any existing medication. Sixty-nine patients (76.7%) were diagnosed as having NTG. Sixty-nine eyes of 69 subjects were enrolled for this prospective, cross-sectional study. They were divided into two groups by age (cut off between 59 and 60 years: group 1, under 60 years; group 2, over 60 years). The correlations between age and the mean, maximum and minimum IOP and IOP variation of the eight nyctohemeral IOP measurements nyctohemerally were evaluated for each group. The times of day at which the maximum and minimum IOPs were observed were recorded. RESULTS: Groups 1 and 2 consisted of 28 patients (age: 50.2+/-6.1 years) and 42 patients (age: 70.3+/-6.1 years) ( P<0.0001), respectively. Neither mean, maximum or minimum IOP nor the IOP variation was significantly different between the groups (all P values > or =0.115). The mean, maximum and minimum IOP in group 2 were all correlated with age ( n=41; r=-0.516, P=0.0004; r=-0.434, P=0.004; r=-0.522, P=0.0004, respectively), while none of those in group 1 was correlated with age (all P values > or =0.303). CONCLUSION: The IOPs of the elder NTG patients have a negative correlation with age while those of the younger ones have no relation to age. PMID- 12545289 TI - Combined penetrating keratoplasty and vitreoretinal surgery with silicone oil: a 1-year follow-up. AB - BACKGROUND: The aim of this study was to assess the functional outcome of eyes with corneal and vitreoretinal diseases following combined surgical procedures (pars plana vitrectomy with temporary keratoprosthesis, vitreoretinal surgery, and penetrating keratoplasty with intravitreal silicone oil tamponade) and to evaluate the factors preserving the clarity of grafts. METHODS: Fifty-three eyes from 49 consecutive patients, operated on between 1991 and 1998 and followed up for at least 12 months, were evaluated retrospectively. The evaluation focused on ocular history, visual acuity (VA), intraocular pressure, anterior and posterior anatomical outcome, and complications. RESULTS: The average follow-up was 28.4 months+/-18.8 months (range 12-84 months); at the final visit 58% of the eyes had better VA, and 73% had equal or better acuity; the cornea remained clear in 68% of the eyes. A decrease in VA was caused by: loss of light perception in 2 eyes, one of which had to be enucleated because of painful phthisis; hypotony or phthisis in 23 eyes; recurrent retinal detachment in 4 eyes that were operated because of trauma; and immunological reaction in 2 eyes. Preoperative factors that contributed to a clear graft, but were not significant, were: lack of trauma (74%), no actual silicone oil filling (78%), preoperative VA of hand movement or better (79%), and attached retina (73%). Further surgery was needed in 55% of cases. The risk of transplant failure was significantly lower in eyes that did not undergo additional surgery (p=0.0001). CONCLUSION: The long-term results of combined surgery with penetrating keratoplasty in eyes that would otherwise be untreatable is often limited by anterior segment complications, mainly secondary graft failure and ciliary body malfunction. For optimal preservation of graft clarity, stable ocular status should be achieved before transplantation to minimize the necessity for further surgery. PMID- 12545290 TI - Visual function and perfusion of the optic nerve head after application of centrally acting calcium-channel blockers. AB - BACKGROUND: In a previous study it was shown that nimodipine 30 mg twice daily leads to an improvement in the visual field in a subgroup of normal-pressure glaucoma patients. To understand the mechanism of action of nimodipine on the visual system, the aim of this study was to examine the influence of nimodipine on different hemodynamic parameters and contrast sensitivity in healthy subjects. METHODS: Thirty-two healthy subjects (21-49 years old, mean age 28 years, 10 male, 22 female) received either nimodipine 30 mg twice a day or a placebo according to the same dosage regimen in a double-blind cross-over study design. The ocular blood flow was measured by means of the ocular blood flow system, the optic nerve head blood flow with the continuous laser Doppler flowmeter (Riva), and contrast sensitivity using the MCT 8000 Multivision Contrast Tester. Measurements were taken at baseline (1T0), 120 min after initial dose (1T3) and after 3 days (3T3) of therapy with 150 mg nimodipine or placebo in total. RESULTS: Contrast sensitivity improved significantly throughout almost all spatial frequencies in the nimodipine-treated subjects ( P=0.01), whereas there was no change in the placebo group. Ocular blood flow and optic nerve head blood flow increased slightly but not significantly in the nimodipine group (1T0: 706.6 microl/min, 9.33 AU; 3T3: 854.3 microl/min, 9.39 AU) and remained unchanged or were even lower in the placebo group ( P>0.05). CONCLUSION: The results showed a significant increase in contrast sensitivity during treatment with nimodipine in healthy subjects. This increase in visual function, however, was not correlated with an increase in ocular or optic nerve head blood flow. Therefore, another mechanism, e.g., a direct effect on the visual system, might be responsible for the improvement in visual function in healthy volunteers under nimodipine therapy. PMID- 12545291 TI - Inter- and intra-observer variability in grading lesions of age-related maculopathy and macular degeneration. AB - PURPOSE: To introduce a revised version of the grading system established by the International ARM Epidemiological Study Group for identifying and quantifying abnormalities of age-related maculopathy (ARM) and age-related degeneration (AMD) and to investigate its reliability, specifically the inter- and intra-observer variability. METHODS: Fifty eyes of 25 patients with ARM or AMD in at least one eye were randomly selected from a large ongoing collection of clinical data and DNA in a tertiary referral UK population. Stereoscopic color fundus photographs were taken with a 30 degrees fundus camera and were centered on the macula. Presence and severity of fundus abnormalities in ARM and AMD were graded using a grid to define macular subfields and standard circles to define the size of lesions. Inter-observer variability was assessed by having three retinal specialists evaluate the color slides and intra-observer variability by re grading the same set. RESULTS: The inter-observer agreement for all subfields was fair to substantial for small hard drusen (70-89%; kappa=0.26-0.63) and intermediate soft drusen (76-94%; kappa=0.27-0.69). Agreement ranged between 87% and 100%, between 50% and 92%, and between 78% and 100% for larger drusen, the presence of hyperpigmentation, and the presence of hypopigmentation, respectively. Agreement was moderate to almost perfect for the presence of geographic atrophy (88-98%; kappa=0.60-0.95) and substantial to almost perfect for the presence of choroidal neovascularization (84-100%; kappa=0.62-1.00). The intra-observer variability for the grading of drusen characteristics and pigmentary changes was similar in magnitude, but slightly greater for features of advanced AMD. CONCLUSION: Reproducibility was achieved using a revised version of the grading system established by the International ARM Epidemiological Study Group. This grading system may therefore be used for phenotyping of ARM and AMD. PMID- 12545292 TI - Neuroretinal basis of visual impairment in the very elderly. AB - PURPOSE: To evaluate the effects of senescence on the morphology and neural function of the retinal nerve fiber layer (NFL) in the very elderly. METHODS: The NFL thickness about the optic nerve head and the neural activity of its neurons were quantified by scanning laser polarimetry and pattern reversal electroretinograms (PERGs), respectively, in a group of young healthy volunteers ( n=27; mean ocular refraction -1.59+/-0.45 D) between 20 and 31 years of age, and a group of healthy elderly volunteers ( n=27; mean ocular refraction +0.11+/ 0.40 D) between 75 and 88 years of age. RESULTS: Relative to the young group, the elderly subjects showed an overall thinning of the NFL of some 22.5%; however, the superior quadrant was attenuated by about 30.0%. The function of the ganglion cell axons in the NFL as reflected by the N95 component of the PERG was reduced by about 28.8% in the elderly compared with the young. CONCLUSION: Aging caused a thinning of the NFL, particularly in the superior quadrant. The evoked potentials from the neurons comprising this layer were attenuated by near equivalent amount. These findings have clinical implications in the differential diagnosis of normal aging versus disease-related alterations in the structure and function of the NFL in the very elderly sector of our population. PMID- 12545293 TI - Exogenous advanced glycosylation end products induce diabetes-like vascular dysfunction in normal rats: a factor in diabetic retinopathy. AB - BACKGROUND: Diabetic retinopathy has been shown to be directly associated with the degree and duration of hyperglycemia, and advanced glycosylation end products (AGEs) have been implicated in this pathological process. The purpose of the experiments reported here was to study the effect of AGE deposition on retinal vascular damage which leads to diabetic retinopathy. METHODS: Intravenous injection of exogenous AGEs was used to treat wild-type non-diabetic Sprague Dawley rats. One of the two retinal slides from each animal was treated using immunohistochemical staining to label retinal vascular AGE deposition, the other H&E staining for counting of capillary pericytes. The results were compared with the findings in untreated wild-type and diabetic controls and in rats treated with unmodified rat serum albumin (RSA). RESULTS: After 2 weeks of continuous treatment, AGEs were identified in the retinal vascular tissue of the AGE-RSA injected group. The average number of retinal capillary pericytes per 10x100 microscope power field was 4.313+/-0.34 (mean +/- SD) in the AGE-RSA-injected group, compared with 5.798+/-0.481 in the control group ( P<0.01). CONCLUSION: These experiments demonstrate that AGEs, independent of other metabolic factors, can induce vascular change resembling that of diabetic retinopathy. PMID- 12545294 TI - Inhibition of experimental angiogenesis of cornea by somatostatin. AB - BACKGROUND: This study evaluated the inhibitory activity of somatostatin 14 on the angiogenesis of cornea in vivo. METHODS: Corneal neovascularization was induced with a pellet containing 90 ng of basic fibroblast growth factor (bFGF) in a rat corneal pocket model. Three kinds of pellets were made containing bFGF plus somatostatin (SST) 0 ng, 20 ng and 200 ng for the control group, group 1 and group 2, respectively. Neovascularization was observed biomicroscopically from day 4 to day 8, and the corneas were then examined for changes in histology. Quantitation of angiogenesis in the cornea was accomplished by caliper and image analysis. RESULTS: The 200-ng dose of SST showed significant inhibition of both length and area of neovascularization on day 7 (0.62+/-0.11 mm vs 1.29+/-0.16 mm, 0.50+/-0.16 mm2 vs 1.35+/-0.29mm2, group 2 vs control; P<0.05). The 20 ng of somatostatin did not demonstrate any significant inhibition of neovascularization compared with the control group. CONCLUSION: Our study demonstrated that SST 14 can reduce bFGF-induced corneal angiogenesis. This shows the potential value of somatostatin in the treatment of corneal neovascularization. PMID- 12545295 TI - Multifocal examination techniques in malingering: case report of a patient with monocular vertical hemianopia. PMID- 12545296 TI - [Brachytherapy after coronary interventions: current state and future perspectives]. AB - Intracoronary brachytherapy is a novel, meanwhile established therapy. It is currently the only interventional procedure which has proven to effectively reduce the restenosis rates after intervention of long and diffuse in-stent restenosis. For this indication, brachytherapy can be regarded as the current treatment of choice. Randomized studies yield promising results for bypass interventions or interventions in small vessels or diabetic patients. These findings may encourage the decision to perform a percutaneous, transluminal intervention in such high-risk patients. In clinical practice, implantation of new stents in combination with brachytherapy procedures should be avoided as far as possible. In any case, the combined antiaggregatory therapy should be conducted sufficiently long to minimize the danger of late stent thrombosis. Under this treatment, the expected thrombosis rates ar within the range of placebo-treated patients. The length of the radiation source should be sufficient to cover the entire interventional injury length to avoid recurrent edge stenosis. De novo lesions are currently not a routine indication for intracoronary brachytherapy. Although intracoronary brachytherapy may effectively reduce restenosis rates in sufficiently irradiated de novo lesion segments, de novo lesions should be treated only within the set-up of controlled studies. The current available data with a follow-up period of up to 5 years show that intracoronary brachytherapy is also in the mid-term a safe and effective therapy for the reduction of restenosis after coronary interventions. PMID- 12545297 TI - Pregnancy in congenital cardiac disease: an increasing challenge for cardiologists and obstetricians -- a prospective multicenter study. AB - OBJECTIVES: Aim of this study was to assess the occurrence of pregnancy-related complications of mother and child during pregnancy, delivery and puerperium in women with CCD prospectively. STUDY DESIGN, POPULATION: This prospective multicenter study included 122 pregnancies in 106 women with CCD (72 with, 34 without previous cardiac surgery). Patient age was 17-44, median 26 years. Cardiac and non-cardiac complications, mode of delivery, abortion, and CCD of the newborn were assessed. RESULTS: Initially all women were in Functional Class I or II. Worsening during pregnancy occurred in 25.5% (n=27), mainly during the second and third trimester. Significant problems due to bleeding, hypertension, rhythm disturbances, endocarditis, liver congestion, increasing cyanosis or death, occurred in 11.3%. Twelve per cent of deliveries were premature. Five women had therapeutic abortion, nine spontaneous abortions, nine preterm births, and one intrauterine death. Seventy-nine per cent (n=85) delivered spontaneously; 21.3% (n=23) had caesarean section. Of the 111 live born children, 5.4% (n=6) had a CCD. CONCLUSIONS: Most women with CCD and a good functional class before pregnancy tolerate pregnancy without major problems. However, pregnancy may induce serious cardiac and obstetric complications. The specific risks require an individualized multidisciplinary patient-management by experienced physicians. PMID- 12545298 TI - [Beta-blocker in secondary prevention of cardiovascular disease]. AB - Since the 1970s beta-blockers are being increasingly used in the treatment of cardiovascular diseases. Many randomized trials have shown a better prognosis of patients under beta-blockade suffering from arterial hypertension and heart failure, after myocardial infarction and left ventricular dysfunction, respectively. Therefore, beta-blockers are an essential part in the treatment of cardiovascular disease. PMID- 12545299 TI - [Feasibility and reliability of a transtelephonic 12 leads ECG]. AB - Feasibility and reliability of a 12 lead electrocardiogram (ECG), recorded by the patient himself and transmitted via telephone are mandatory for prehospital diagnosis of myocardial ischemia in patients with coronary heart disease. In this study, a 12-lead ECG recorded by patients and transmitted to the cardiology call center via telephone (Tele-ECG; model CG-7100; Card Guard) was compared with the conventional 12-lead ECG from the same patient recorded at the same time in 128 cases. The Tele-ECGs received by the call center were compared with conventional ECG by two cardiologists and one internist independently and blindly. In relation to the conventional ECG, reproducibility of PQ, QRS and QT-durations [ms] as measured in the Tele-ECG was 85%. Concordance between Tele-ECG and conventional ECG in the detection of negative T-waves was very high (Kappa value (kappa) 0.94, 0.96 and 0.97), respectively, depending on the physician. ST-segment changes were diagnosed correctly in most of the cases in Tele-ECG (kappa=0.98-0.99), as compared with the conventional ECG by all physicians. Concordance between Tele- and conventional ECG concerning the pattern of old myocardial infarction was very high (kappa=0.99) for all infarct localization. The limitations of Tele-ECG were of technical nature. In 14 cases, the peripheral electrodes were displaced; in 12 cases, there were baseline artifacts. These technical errors could be corrected in 126/128 patients (98.4%) by transtelephonic verbal communication. In conclusion, the reliability and feasibility of the 12-lead Tele-ECG-recorder used in this pilot study could be demonstrated. In an ongoing randomized controlled multicenter study, it is being investigated whether the strategy of prehospital diagnosis of acute coronary syndromes by Tele-ECG in combination with verbal communication is able to reduce pre- and in-hospital time delay, to avoid unnecessary hospital admissions and to reduce the economic burden of coronary heart disease. PMID- 12545300 TI - [Predicting the risk of atrioventricular conduction lesions after percutaneous septal ablation for obstructive hypertrophic cardiomyopathy]. AB - BACKGROUND AND INTRODUCTION: Damage to the AV conduction system is a frequent complication of percutaneous septal ablation (PTSMA) that needs early and reliable identification of those patients (pts.) at risk for complete heart block (CHB) and subsequent pacemaker implantation. METHODS AND RESULTS: In the first 39 pts. who underwent PTSMA in 1996, AV conduction recovery needed up to 11 days. One pt. suffered from unexpected CHB after 9 days. Seven pts. who needed a DDD pacemaker (DDD-PM) were compared to those without conduction disturbances. A score was established which identified all DDD-PM candidates retrospectively if they presented with >12 score points. In the following 137 consecutive pts. treated in 1997 and 1998, this score was applied prospectively, and again correctly identified all candidates for a DDD-PM. In addition, a low risk group was identified with <8 score points. From 1999 on, the score was applied in routine clinical decision-making in 120 consecutive pts. with respect to DDD-PM implantation. All low risk pts. (<8 points) remained free from bradycardias, while 2/54 pts. (4%) of the intermediate risk group, and 20/23 pts. (87%) of the high risk group had to undergo DDD-PM implantation. Pts. with a first-degree AV block or those with a right bundle branch block at baseline had no excess risk, while 50% of the pts. with a left bundle branch block (LBBB) needed a DDD-PM. CONCLUSIONS: Based on pre-interventional data and careful monitoring of the first 48 hours after PTSMA, identification of pts. at risk for CHB and subsequent DDD PM implantation seems to be possible. Pts. with a score <8 seem to be at low, those with >12 points at high risk. In the remaining cases watchful waiting with prolonged monitoring may allow AV conduction to recover, thus, reducing the number of unnecessary DDD-PM implantations. In cases with LBBB at baseline, however, implantation of a DDD-PM should be considered first-line therapy. PMID- 12545301 TI - [Stent implantation as therapy of first choice in adults with coarctation]. AB - Stent implantation for coarctation of the aorta is an alternative to surgery or balloon dilation. We report our results in 12 patients with a median age of 22 years (10 to 28 years) and a body weight of 60 kg (32 to 97 kg). Nine patients had native stenosis and three had recoarctation after surgery. Invasively measured systolic pressure gradients ranged from 20 to 100 mmHg. Nine patients suffered from brachiocephalic hypertension. Eleven implantations were successful with a median dilatation of 17 mm (15-25 mm). Residual gradients were 0-5 mmHg in seven patients, 5-10 mmHg in three and 15 mmHg in one patient with postoperative recoarctation. Twenty-one months (2-37 months) after intervention, no hemodynamically relevant intimal proliferations, no restenosis, and no aneurysms were present. Thus, stent implantation is a very promising therapy for coarctation of the aorta in adults and is on its way to becoming the therapy of first choice. PMID- 12545302 TI - Decellularized pulmonary homograft (SynerGraft) for reconstruction of the right ventricular outflow tract: first clinical experience. AB - INTRODUCTION: Cryopreserved homograft valve conduits have been used to reconstruct the right and left ventricular outflow tract. Long-term studies have shown homograft degeneration and calcification, and it has been postulated that immunological mediated phenomena in a manner similar to that seen in chronic rejection may contribute to the degeneration process. The development of a decellularized, non-glutaraldehyde-fixed valve conduit creates a non-immunogenic connective tissue matrix for autologous recellularization by host cells. The aim of the study was to characterize the clinical and hemodynamic pattern in human implants of the novel decellularized pulmonary homografts (SynerGraft). METHODS: Reconstruction of the right ventricular outflow tract was performed in 17 patients: 15 patients with aortic valve disease and the Ross procedure, and two patients with redo procedures following Fallot tetralogy and severe pulmonary regurgitation. Patients with the Ross procedure with standard cryopreserved homografts as neopulmonic conduits served as controls. Within the follow-up over six months morphological and hemodynamic parameters were characterized by echocardiography: maximal and mean pressure gradient across the right and left ventricular outflow tract, their effective orifice areas, determination of neopulmonic and neoaortic regurgitation. RESULTS: One patient died six weeks following surgical treatment due to non-valve related end-stage cardiopulmonary failure; all patients were free of valve-related complications during the follow up period. The matched Ross patients showed a gradual but significant increase of both the maximal and mean pressure gradient across the right ventricular outflow tract (Delta P max 5.5+/-2.5 to 11.4+/-6.4 mmHg, p=0.002; Delta P mean 3.0+/-1.3 to 6.2+/-3.9 mmHg, p=0.003), whereas in the SynerGraft group increase of pressure gradients were measurable but did not reach statistical significance (Delta P max 7.1+/-3.7 to 10.1+/-3.9 mmHg, p=0.11; Delta P mean 3.6+/-1.6 to 5.5+/-2.3 mmHg, p=0.12). The pulmonary effective orifice areas decreased in the control group from 1.74+/-0.33 to 1.18+/-0.36 cm(2)/m(2) (p=0.001). Within the SynerGraft group time dependent reduction of the orifice area was significantly less (1.51+/-0.37 to 1.25+/-0.26 cm(2)/m(2); p=0.08). CONCLUSION: Up to six months after implantation reconstruction of the right ventricular outflow tract with decellularized homografts was safe, stable, and the morphological and hemodynamic features are promising. PMID- 12545303 TI - [Calcifications in temporal arteries -- their morphogenesis in comparison to physiological osteogenesis]. AB - OBJECTIVE: Vascular calcification, traditionally regarded as a dystrophic process, has recently been interpreted as a bone-like biologically regulated phenomenon. Because temporal arteries which also contain calcifications are easily available from biopsies of older individuals with suspected giant cell arteritis, we studied the morphogenesis of this calcification in comparison with the development of fetal bone. MATERIAL AND METHODS: Formaldehyde fixed arteries were processed in paraffin sections and investigated by light-, transmission-, and scanning electron microscopy. The atomic composition of the calcifications was estimated by X-ray microanalysis. Fetal bone, also fixed with formaldehyde, was investigated using identical methods. RESULTS: Early calcifications are often present as focal or diffuse granular mediacalcinosis. With increasing age, calcified granules appear in association with the internal elastic membranes and progress to sheet-like calcifications. Ultrastructurally calcospherites, not rarely exhibiting the Liesegang phenomenon, are the hallmark of early calcifications. In advanced sheet-like calcification calcospherites could be detected as components of the calcified lumps. Calcospherites also appear in endochondral calcification, but are absent in calcified osteoid. The ultrastructure of calcospherites in provisional endochondral ossification differs from that in vascular calcification. Instead of corpuscles as with the Liesegang phenomenon, radial clusters of needle-like crystals resembling apatite are present. While calcospherites of the tunica media often contain a high amount of magnesium, calcified sheets as well as bone exhibited in contrast a low magnesium content. CONCLUSIONS: It is concluded that, comparable to granular mediacalcinosis of the aorta, development of calcified arteriosclerotic plaques, and Monckeberg's disease, calcification of small muscular temporal arteries is also initially a calcospherite-dependent process. It is generally accepted that these calcified corpuscles represent remnants of calcified necrotic or apoptotic cells. In the phase of confluence with the appearance of sheet-like calcification, a low bone-like magnesium content that differs from the high amount in the isolated medial calcospherites was measured. This finding, together with the observation that calcified sheets contain calcospherites, supports the hypothesis that vascular calcification in general is different from bone formation and has to be regarded as a biphasic process initiated by the appearance of calcospherites and followed by a secondary calcifying phase with the formation of bone-like apatite that leads to rock hard lumps of calcified vessels. PMID- 12545304 TI - Plasma catecholamines and N-terminal proBNP in patients with acute myocardial infarction undergoing primary angioplasty. Relation to left ventricular function and clinical outcome. AB - BACKGROUND: Neither profiles nor prognostic values of neurohormonal markers have been prospectively evaluated in patients with acute myocardial infarction (AMI) undergoing primary angioplasty. METHODS AND RESULTS: In 118 consecutive patients with AMI undergoing successful reperfusion (TIMI 2 and 3) by primary angioplasty, plasma concentrations of norepinephrine, epinephrine and N-terminal proBNP (NT proBNP) were measured before, 60 min and 10 days after angioplasty. Catecholamine concentrations (mean+/-SEM) rose to a maximum in the first hour after angioplasty (norepinephrine: 602+/-44 ng/L, epinephrine: 213+/-24 ng/L) and returned to normal at day 10. Conversely, NT-proBNP levels maintained a further increase from 799+/-44 pmol/L at baseline to 924+/-54 pmol/L at day 10. A NT-proBNP concentration above median at 60 min post-angioplasty predicted major adverse cardiac events (n=27) during the 18-36 month follow-up with an odds ratio of 5.9 (1.7-20.3) and was superior to catecholamines, to left ventricular ejection fraction and to other established postinfarction risk markers. CONCLUSIONS: In a low-risk cohort of patients with AMI undergoing successful reperfusion therapy, plasma NT-proBNP concentrations are elevated for at least ten days. The prognostic value of early plasma NT-proBNP should be further evaluated concerning its ability to facilitate risk stratification of infarct patients. PMID- 12545305 TI - A coconut left atrium 23 years after mitral valve replacement for chronic mitral stenosis. AB - We present the case of a 65 year-old female who was admitted to the hospital because of deterioration of chronic dyspnea. Twenty-three years prior to this admission, mitral valve replacement for chronic mitral stenosis was performed using a Starr-Edwards caged-ball prosthesis. There was severe pulmonary hypertension. On transthoracic echocardiography and on cineradiography, the function of the mitral valve prosthesis was unimpaired. However, cineradiography showed extensive mural calcification of the left atrium; we report this remarkable finding. PMID- 12545306 TI - [Percutaneous stents for the treatment of stenosis of descending aorta]. AB - Stenotic processes of the descending aorta lead to a transstenotic gradient with either risk of proximal hypertension especially affecting the cerebral circulation combined with the risk of distal malperfusion. This article describes the technique of percutaneous stenting of suprarenal aortic stenosis in two patients with different genesis of aortic obstruction in whom operative correction was refused due to elevated surgical risk. The first patient (female, 38 years) suffered from aortic stenosis at the thoracoabdominal level revealing a residual lumen of 3 mm. The second patient (male, 71 years) had a chronic type B aortic dissection and developed dynamic compression of true lumen and thus peripheral malperfusion. Both patients were treated successfully with percutaneous implantation of self-expanding stents. During the follow-up of 6 and 10 months, respectively, both patients were free of any symptoms. The technique of percutaneous stenting of static and dynamic stenotic processes of the aorta led to excellent mid-term results. Long-term results of large patient cohorts are not available yet; however, all patients subjected to such a palliative procedure should be followed in structured registries in an effort to standardize the concept and develop therapeutic recommendations. PMID- 12545307 TI - Severe heart failure from light chain cardiomyopathy (cardiac amyloidosis). AB - Primary (AL)amyloidosis is characterized by clonal production of immunoglobulin with subsequent deposition in several organs. We describe the clinical features of a 66-year old female who was referred to our department for congestive heart failure. One year before, she was examined and found to have diastolic dysfunction of the left ventricle. We could evaluate the diagnosis of primary amyloid cardiomyopathy by echocardiography, Doppler echocardiography and laboratory findings and confirmed the diagnosis by biopsy of the rectum mucosa. Clinical outcome was poor, because therapy is poor. PMID- 12545308 TI - [Policy paper to the cardiac re-sychronization therapy]. PMID- 12545309 TI - Viral lysis, flagellate grazing potential, and bacterial production in Lake Pavin. AB - Abundances of different compartments of the microbial loop (i.e., viruses, heterotrophic bacteria, nonpigmented nanoflagellates, and pigmented nanoflagellates), bacterial heterotrophic production (BHP), viral lysis, and potential flagellate grazing impacts on the bacterial assemblages were estimated during a short-term study (24 h) conducted in June 1998 in the epilimnion (5 m) and metalimnion (10 m) of a moderate-altitude oligomesotrophic lake (Lake Pavin, France). Viral and bacterial abundances were higher in the metalimnion than in the epilimnion, whereas pigmented and nonpigmented nanoflagellates were more numerous in the epilimnion. The control of the BHP due to viral lysis (determined by examination of viral-containing bacteria using a transmission electron microscope) was significantly higher in the meta- (range = 6.0-33.7%, mean = 15.6%) than in the epilimnion (3.5-10.3%, 6.4%). The same was for the losses of BHP from the potential predation by nanoflagellates which ranged from 0.5 to 115.4% (mean = 38.7%) in the epilimnion, and from 0.7 to 97.5% (mean = 66.7%) in the metalimnion. Finally, estimated viral mediated mortality rates from the percentage of visibly infected cells and potential nanoflagellate grazing rates based on assumed clearance rates suggest that flagellates consumed a larger proportion of bacterial production than was lost to viral lysis. PMID- 12545310 TI - Arbuscular mycorrhizal symbiosis changes the colonization pattern of Acacia tortilis spp. Raddiana rhizosphere by two strains of rhizobia. AB - The aim of the study was to assess the effect of the mycorrhizosphere of A. tortillis spp. raddiana mycorrhized with Glomus intraradices on the root nodulation by Sinorhizobium terangae (ORS 1009) and/or Mesorhizobium plurifarium (ORS 1096) in two different culture substrates (sandy soil and sand). The endomycorrhizal fungus only stimulated plant growth in the sandy soil. Moreover, arbuscular mycorrhizal infection enhanced the nodulation process in both culture substrates. Beside the stimulatory effects of the mycorrhizosphere on both rhizobia development, fungal symbiosis induces two different dynamics of each bacterial strains in the sand-grown plants. These results suggest specific relationships could occur during the development of the tripartite symbiosis, at physiological and molecular level. From a practical point of view, the role of arbuscular mycorrhizas in improving nodulation and N2 fixation is universally recognized. The fungal symbiosis could modify the development of bacterial inoculants along the root systems. This effect is of particular interest in the controlled inoculation of selected rhizobia. PMID- 12545311 TI - Mycorrhizal and root endophytic fungi of containerized Picea glauca seedlings assessed by rDNA sequence analysis. AB - Fungi colonizing fine roots of containerized Picea glauca seedlings were assessed in four large conifer nurseries in northern Alberta. PCR amplification of fungal rDNA (internal transcribed spacer and a portion of the 5' end of the large subunit gene) from random samples of fine feeder roots gave between 1 and 4 amplicons per seedling. Amplicons were either separated by electrophoresis and sequenced directly, or cloned and sequenced. The resulting sequences were compared to sequences obtained from cultures established from seedling roots and from GenBank by maximum parsimony analysis. ITS sequences formed 11 distinct clades, each including at least one reference sequence. The ectomycorrhizal basidiomycetes Thelephora americana and Amphinema byssoides were dominant, whereas ascomycetes were less common. Fungi with sequences similar to members of the Heleotiales which form ericoid mycorrhizas were also present. Correspondence analysis revealed strong positive and negative associations among fungal taxa as well as an influence of applied fertilizer level on fungal diversity and species composition. PMID- 12545312 TI - Sunscreen products increase virus production through prophage induction in marine bacterioplankton. AB - Classical pollutants (e.g., hydrocarbon, pesticides) have been recently recognized to induce lytic cycle in lysogenic bacteria, but information on micro pollutants is almost completely lacking. We investigated the effects of cosmetic sun products (sunscreen and solar oil) on viral abundance and bacterial activity. We found that both sunscreen and solar oil acted as pollutants, inducing viral development and controlling bacterial abundance and production, thus leading to an increase of the virus to bacterium ratio. Short-term experiments revealed that sunscreen supplementation induced the lytic cycle in a large fraction of total bacterial abundance (13-24% of bacteria, at low and high concentrations, respectively), whereas solar oil had a lower impact (6-9%). A synchronized development of the phage-host system was observed only after sunscreen addition. The addition of sunscreen, even at low concentrations, had a significant impact on all enzymatic activities (aminopeptidase, glucosidase, and phosphatase), which increased significantly. However, when enzymatic activities were normalized per cell, a selective enhancement was observed for certain enzymes (e.g., aminopeptidase) and inhibition for others (e.g., glucosidase). These results indicate that sunscreen products can modify C, N, and P biogeochemical cycling in seawater and increase virus abundance through prophage induction in marine bacterioplankton. PMID- 12545313 TI - Microbial diversity and resistance to copper in metal-contaminated lake sediment. AB - Contamination of habitats with heavy metals has become a worldwide problem. We describe herein the analysis of lake sediment contaminated with high concentrations of copper as a consequence of mine milling disposal over a 100 year period. Copper concentrations in the sediment were found to vary with depth and ranged from 200 to 5500 ppm. Analysis of the microbial community with T-RFLP identified a minimum of 20 operational taxonomic units (OTU). T-RFLP analysis along a depth profile detected as many as nine shared OTUs across 15 centimeters, suggesting a conservation of community structure over this range. Only two genera, Arthrobacter and Ralstonia, were detected among 50 aerobic copper resistant isolates cultivated on R2A, one of which (Ralstonia sp.) was characterized by the sequestration of copper, identified by electron diffraction scanning, in growing colonies. Scanning electron microscopy showed changes to the outer envelope of the cells when grown in the presence of copper. The copper resistant Ralstonia isolates were also resistant to Ni, Cd, and Zn, showing two patterns of phenotypic resistant to these three metals in which either resistance to Zn or Ni was expressed in an isolate but never both. PMID- 12545315 TI - Antagonism between bacteria and fungi on decomposing aquatic plant litter. AB - Bacterial and fungal decomposers of aquatic plant litter may exhibit either synergistic or antagonistic interactions, which are likely to influence microbial growth as well as the decomposition of litter and, eventually, the carbon metabolism of aquatic systems. To elucidate such interactions, we inoculated decomposing Phragmites culms in microcosms with fungal isolates and with natural communities of bacteria and fungi in different combinations. The development of fungal and bacterial biomass and the carbon dynamics were studied during several months of degradation. The results show a bilateral antagonistic relationship between bacteria and fungi. After 3 months, fungal biomass accumulation was approximately 12 times higher in the absence than in the presence of bacteria. Bacterial biomass accumulation was about double in the absence of fungi compared to when fungi were present. Similar interactions developed between a natural assemblage of bacteria and five different fungal strains isolated from Phragmites litter (three identified hyphomycetes and two unidentified strains). Despite the great difference in biomass development between the treatments, the carbon metabolism was similar regardless of whether fungi and/or bacteria were present alone or in coexistence. We suggest that the antagonism between bacteria and fungi is an important controlling factor for microbial colonization and growth on aquatic plant litter. PMID- 12545314 TI - Residual impact of the biocontrol inoculant Pseudomonas fluorescens F113 on the resident population of rhizobia nodulating a red clover rotation crop. AB - A field trial was previously conducted in which sugarbeet seeds were either untreated, inoculated with the biocontrol strain Pseudomonas fluorescens F113Rif, or treated with chemical fungicides. Following harvest of sugarbeet, the field site was sown with uninoculated red clover. The aim of this study was to assess the residual impact of the microbial inoculant (and the fungicide treatment) on the diversity of resident rhizobia nodulating the red clover rotation crop. The percentage of nodules yielding rhizobial isolates after surface disinfection was 67% in the control and 70% in the P. fluorescens F113Rif treatment, but only 23% in the chemical treatment. Isolates were characterized by RAPD analysis. The main RAPD cluster (arbitrarily defined at 70% similarity) was prevalent in all three treatments. In addition, the distribution of RAPD clusters followed a log series model, regardless of the treatment applied, indicating that neither the microbial inoculant nor the fungicide treatment had caused a strong perturbation of the rhizobial population. When the P. fluorescens F113Rif and control treatments were compared using diversity indices, however, it appeared that the genetic diversity of rhizobia was significantly less in the inoculated treatment. The percentage of rhizobia sensitive to 2,4-diacetylphloroglucinol (Phl; the antimicrobial metabolite produced by P. fluorescens F113Rif) fluctuated according to field site heterogeneity, and treatments had no effect on this percentage. Yet, the proportion of Phl-sensitive isolates in the main RAPD cluster was lower in the P. fluorescens F113Rif treatment compared with the control, raising the possibility that the residual impact of the inoculant could have been partly mediated by production of Phl. This impact on the rhizobial population took place without affecting the functioning of the Rhizobium-clover symbiosis. PMID- 12545316 TI - Survival of filamentous fungi in hypersaline Dead Sea water. AB - A variety of filamentous fungi have recently been isolated from the Dead Sea (340 g/L total dissolved salts). To assess the extent to which such fungi can survive for prolonged periods in Dead Sea water, we examined the survival of both spores and mycelia in undiluted Dead Sea water and in Dead Sea water diluted to different degrees with distilled water. Mycelia of Aspergillus versicolor and Chaetomium globosum strains isolated from the Dead Sea remained viable for up to 8 weeks in undiluted Dead Sea water. Four Dead Sea isolates (A. versicolor, Eurotium herbariorum, Gymnascella marismortui, and C. globosum) retained their viability in Dead Sea water diluted to 80% during the 12 weeks of the experiment. Mycelia of all species survived for the full term of the experiment in Dead Sea water diluted to 50% and 10% of its original salinity. Comparison of the survival of Dead Sea species and closely related isolates obtained from other locations showed prolonged viability of the strains obtained from the Dead Sea. Spores of isolates obtained from the terrestrial shore of the Dead Sea generally proved less tolerant to suspension in undiluted Dead Sea water than spores of species isolated from the water column. Spores of the species isolated from the control sites had lost their viability in undiluted Dead Sea water within 12 weeks. However, with the exception of Emericella spores, which showed poor survival, a substantial fraction of the spores of Dead Sea fungal isolates remained viable for that period. The difference in survival rate between spores and mycelia of isolates of the same species points to the existence of adapted halotolerant and/or halophilic fungi in the Dead Sea. PMID- 12545317 TI - Reversible symptomatic dilated cardiomyopathy in older children and young adolescents due to primary non-sinus supraventricular tachyarrhythmias. AB - Dilated cardiomyopathy (DCM) due to a primary supraventricular tachycardia not originating from the sinus node is not frequently seen in older children or adolescents. However, it is important to recognize this entity as a reversible cause of DCM to avoid costly and inappropriate treatments for these patients. We describe 7 patients who presented with DCM. Five were misdiagnosed as having "sinus" tachycardia secondary to an idiopathic DCM, and 2 were correctly diagnosed as having DCM secondary to an atrial tachycardia. All underwent electrophysiologic treatment of the tachycardia with remission of the DCM. PMID- 12545318 TI - Use of low-dose ketamine and/or midazolam for pediatric cardiac catheterization. AB - Ketamine and midazolam are commonly used in children undergoing cardiac catheterization. However, there is controversy regarding the safety of administering these agents in the absence of an anesthesiologist. We retrospectively reviewed pediatric cardiac catheterization procedures at our institution between 1996 and 1997. A total of 154 patients (0.3-192 months) underwent a total of 205 procedures. They received ketamine (n = 79, 1.05 +/- 0.88 mg/kg/hr), midazolam (n = 35, 0.14 +/- 0.09 mg/kg/hr), or both (n = 91; ketamine, 1.13 +/- 0.84 mg/kg/hr; midazolam, 1.57 +/- 1.03 mg/kg//hr). In 18.5% of patients there were complex cardiac lesions. Mean procedure time was 79 +/- 36.2 minutes. Pre- and postprocedure systolic and diastolic mean blood procedure 72 +/- 14 and 68 +/- 12 mmHg, respectively. Pre- and postprocedure O2 saturation was 93.19 +/- 8.72 and 93.63 +/- 8.3, respectively. One patient required intubation, and 15% required oxygen therapy. The mortality rate was zero. The anesthesiologist's assistance was requested by the cardiologist in 21 procedures (group A) and not requested in 184 procedures (group B). The two groups were not different in relation to the drug used (p = 0.283) or the complexity of the cardiac lesions (p = 0.051). However, there was significant difference between the two regarding the need for supporting drugs (3/21 vs 3/184, p = 0.02) or oxygen treatment (7/21 vs 26/184, p = 0.014). No patients in group B required intubation, whereas 14% and 1.6% required oxygen therapy and supporting drugs, respectively. We conclude that low-dose ketamine and midazolam can be administered safely to most pediatric patients by the cardiologist, who can safely predict the need for an anesthesiologist. PMID- 12545319 TI - Percutaneous hydrodynamic thrombectomy for femoral arterial thrombosis after arterial catheterization. AB - We report on a 3-year-old boy in whom a complete occlusive thrombus in the right femoral artery was removed rapidly and successfully using a hydrodynamic thrombectomy catheter. There was no bleeding complication, reocclusion, vascular injury, or loss of motor or sensory function. Hydrodynamic thrombectomy can be an important part of the treatment of thrombosis in children. PMID- 12545320 TI - Myocardial perfusion and coronary flow reserve assessed by positron emission tomography in patients after Fontan-like operations. AB - Ventricular dysfunction in patients after Fontan-like operations (FLOs) is a serious complication that might contribute to poor long-term results. Ischemic heart disease will have debilitating consequences on a Fontan heart. Ten patients (15.8 +/- 5.01 years) after FLO had transesophageal echocardiography and cardiac catheterization 9.3 +/- 4.2 years after surgery. Myocardial perfusion was assessed by NH3-positron emission tomography (rest/adenosine) and compared with that of 10 healthy adults (26.1 +/- 6.3 years). Ventricular function was normal in 4 and reduced in 6 patients; end systolic and end diastolic meridional wall stress was significantly elevated in the FLO group. Coronary angiography revealed no stenosis of the coronaries. Compared to normals, myocardial blood flow (MBF) at rest was higher in the FLO group (0.99 +/- 0.25 vs 0.77 +/- 0.17 ml/g/min, p <0.05), whereas MBF after vasodilatation (2.12 +/- 0.78 vs 3.10 +/- 0.85 ml/g/min, p <0.05) and coronary flow reserve (CFR) was reduced (2.5 +/- 0.88 vs 4.1 +/- 1.01, p <0.05), especially in those with impaired ventricular function. Coronary vascular resistance after vasodilatation was elevated in the FLO group (38.2 +/- 17.4 vs 24.5 +/- 8.3 mmHg/ml/g/min, p <0.05). Altered MBF, increased meridional wall stress, and impaired CFR are common findings in FLO. Attenuated CFR and reduced ventricular function are significantly correlated and may be risk factors for the long-term outcome. PMID- 12545325 TI - Transhepatic catheterization using ultrasound-guided access. AB - The efficacy and safety of ultrasound guidance to obtain transhepatic access for cardiac catheterization were investigated in this study. The transhepatic route for access to perform cardiac catheterization has become an acceptable alternative when conventional routes of access have failed. However, the use of ultrasound to guide transhepatic access has not been reported in the literature. We performed a retrospective chart review. Patient characteristics, indications for catheterization, procedures performed, and complications were recorded. All patients who underwent transhepatic cardiac catheterization at Duke University Medical Center were included in this study. Eight patients underwent 12 catheterizations. The median age was 5.3 years (range, 9 months to 13 years) and median weight 18.7 kg (range, 7.1-44.8 kg). Seven catheterizations were diagnostic and 5 were interventional. There were no complications. Transhepatic access with ultrasound guidance is a safe and effective option for obtaining venous access for cardiac catheterization. PMID- 12545326 TI - Total anomalous pulmonary venous return to the azygous vein: an unusual presentation in an infant. PMID- 12545327 TI - Air Embolism following CAT scan in a patient with hypoplastic left heart syndrome. PMID- 12545328 TI - Evidence of cross-tolerance between behavioural effects of nicotine and cocaine in mice. AB - RATIONALE: Studies have reported that chronic exposure to nicotine does not alter the effects of cocaine on locomotor activity, and vice versa. However, the apparent lack of effect of one drug on the behavioural response to the other may be due to an exclusive focus on locomotor activity as the target behaviour. OBJECTIVE: To test whether repeated pretreatment with nicotine causes tolerance or sensitization to cocaine's effects on diverse behaviours: locomotion, rearing, grooming, and immobility. Similarly, the effects of repeated cocaine treatment on the acute response to nicotine were also tested. METHODS: Mice were pretreated with 14 injections of nicotine (0.3 mg/kg), cocaine (5 mg/kg) or saline, the injections being given once daily, except for three breaks of two days each. Two days after the final pretreatment injection, mice were given a challenge injection of saline, cocaine (3 or 5 mg/kg) or nicotine (0.3 or 1 mg/kg), and observed in a large test cage for 40 min using a time-sampling procedure. RESULTS: Repeated administration of either drug produced some tolerance to subsequent challenge with the same dose of the drug. Prior nicotine exposure significantly attenuated cocaine-induced decreases in grooming and increases in rearing, but did not significantly affect other behaviours. In contrast, prior cocaine exposure failed to alter nicotine's effects on any behaviour. CONCLUSIONS: Cross-tolerance between nicotine and cocaine (but not vice-versa) can be demonstrated if several behaviours are observed; measures of locomotor activity are less sensitive to the effect. The asymmetrical pattern of cross tolerance may be due to differential inhibition of dopamine uptake by the two drugs. PMID- 12545329 TI - Dissociation in the modulatory effects of environmental novelty on the locomotor, analgesic, and eating response to acute and repeated morphine in the rat. AB - RATIONALE: We have previously shown that environmental novelty can potentiate the activating effects of morphine and the development of sensitization to this effect. OBJECTIVES. Our main goal was to determine whether environmental novelty can also modulate the prophagic (time spent eating and food intake; experiment 1) and/or the analgesic (tail-flick test; experiments 2 and 3) effect of morphine, as well as the development of tolerance or sensitization to these effects. METHODS: In experiment 1, two groups of rats were administered seven intraperitoneal (i.p.) injections of either saline or morphine (4.0 mg/kg) either in their home cages (home groups) or in a distinct environment (novelty groups). After 7 days of withdrawal, both groups underwent a morphine challenge (4.0 mg/kg, i.p.). In experiment 2, home and novelty rats were administered four doses of morphine (0.0, 2.0, 4.0, and 8.0 mg/kg, i.p.) following a counterbalanced order. In experiment 3, home and novelty rats were administered eight intraperitoneal (i.p.) injections of either saline or morphine (8.0 mg/kg) and then given a morphine challenge (4.0 mg/kg). RESULTS: Environmental novelty enhanced the locomotor activating effect of morphine and the expression of sensitization to this effect (even after a period of withdrawal). Environmental novelty had relatively little effect on morphine-induced eating, and no effect on morphine-induced analgesia. CONCLUSIONS: Environmental context can have very different consequences on distinct drug effects as well as on distinct neurobehavioral adaptations to the same drug treatment (e.g., psychomotor sensitization versus analgesic tolerance). PMID- 12545330 TI - Cognitive rehabilitation in schizophrenia: a quantitative analysis of controlled studies. AB - RATIONALE: Cognitive rehabilitation is now recognized as an important tool in the treatment of schizophrenia, and findings in this area are emerging rapidly. There is a need for a systematic review of the effects of the different training programs. OBJECTIVES: To review quantitatively the controlled studies on cognitive rehabilitation in schizophrenia for the effect of training on performance on tasks other than those practiced in the training procedure. METHODS: A meta-analysis was conducted on 12 controlled studies of cognitive rehabilitation in schizophrenia taking into account the effects of type of rehabilitation approach (rehearsal or strategy learning) and duration of training. RESULTS: The mean weighted effect size was 0.45, with a 95% confidence interval from 0.26 to 0.64. Effect sizes differed slightly, depending on rehabilitation approach, in favor of strategy learning, but this difference did not reach statistical significance. Duration of training did not influence effect size. CONCLUSIONS: Cognitive rehabilitation can improve task performance in patients with schizophrenia and this effect is apparent on tasks outside those practiced during the training procedure. Future studies should include more real world outcomes and perform longitudinal evaluations. PMID- 12545331 TI - Electrical and chemical stimulation of the basolateral complex of the amygdala reinstates cocaine-seeking behavior in the rat. AB - RATIONALE: The basolateral complex of the amygdala (BLC) is part of a neural circuit that is activated in humans during cocaine craving elicited by exposure to drug-related environmental cues. In animals, the BLC is necessary for cocaine seeking behavior elicited by cocaine-associated cues. It has not been determined whether BLC activation is sufficient to reinstate cocaine seeking. OBJECTIVES: To determine whether electrical or excitatory amino-acid stimulation of the BLC is sufficient to induce reinstatement of cocaine seeking in rats. METHODS: Rats were catheterized and trained to lever-press for intravenous cocaine infusions on a fixed ratio (FR)-1 schedule of reinforcement. Once baseline cocaine-taking criteria were met, lever-pressing behavior was extinguished by substitution of saline for cocaine. After meeting criteria for extinction, animals were subjected to brief electrical (20 Hz, 400 microA or 2 Hz, 400 microA; 200 pulses per stimulation) or N-methyl- d-aspartate (NMDA; 250 ng/0.5 microl) BLC stimulation and lever pressing behavior was monitored. RESULTS. Electrical BLC stimulation with 20-Hz produced reinstatement of lever pressing previously associated with cocaine self-administration, while 2-Hz stimulation did not. Electrical stimulation of cerebellar and medial forebrain bundle loci did not reinstate cocaine seeking. Hence, the reinstatement was frequency dependent and anatomically selective. NMDA microinjections into the BLC also reinstated cocaine seeking behavior. CONCLUSIONS: BLC stimulation is sufficient to reinstate cocaine seeking behavior in the rat. These results are congruent with the hypothesis that the basolateral complex of the amygdala is part of a neural system mediating drug seeking behavior. PMID- 12545332 TI - Altered pain sensitivity and morphine-induced anti-nociception in mice lacking CCK2 receptors. AB - RATIONALE: Cholecystokinin (CCK) interacts with the endopioid system in the regulation of various physiological functions, including the control of pain sensitivity, motor activity and emotional behaviour. OBJECTIVE: The aim of the present work was to study the pain sensitivity, morphine-induced antinociception and density of opioid receptors in mice lacking CCK(2) receptors. METHODS: Plantar analgesia and hotplate tests were used to evaluate pain sensitivity and morphine-induced antinociception. The parameters of opioid receptors were analysed by using [(3)H]-diprenorphine binding. RESULTS: In the plantar analgesia test the latency of hind paw withdrawal was significantly increased in CCK(2) receptor deficient mice compared to wild-type (+/+) littermates. The treatment with saline reversed the reduced pain sensitivity in heterozygous (+/-) and homozygous (-/-) mice. The administration of morphine (1 mg/kg) induced a significantly stronger antinociceptive effect in homozygous (-/-) mice compared with wild-type (+/+) animals. In the hotplate test, only homozygous (-/-) mutant mice displayed the delayed latency of hind paw licking/shaking in comparison with wild-type (+/+) mice. The injection of saline and isolation of mice for 30 min reversed the delayed response in homozygous (-/-) mice. However, in this test, the anti-nociceptive action of morphine (5-10 mg/kg) in mutant mice did not differ from that in wild-type (+/+) littermates. By contrast, the jump latency was decreased in both homozygous (-/-) and heterozygous (+/-) mice in the hotplate test. The increased density of opioid receptors was established in the striatum of homozygous (-/-) mice. CONCLUSION: It is apparent that the targeted mutagenesis of the CCK(2) receptor gene has different effects on the sensitivity of opioid receptors in various brain structures. This is a probable reason for the altered pain sensitivity and morphine-induced antinociception in mutant mice compared to wild-type (+/+) littermates. PMID- 12545333 TI - Adaptive changes in the rat hippocampal glutamatergic neurotransmission are observed during long-term treatment with lorazepam. AB - RATIONALE: Chronic treatment with benzodiazepines induces tolerance to most of their pharmacological effects. The best-studied neurochemical correlation to this phenomenon involves GABAergic adaptive changes. However, some compensation by excitatory neurotransmission could also be postulated. OBJECTIVE: The aim of this work was to investigate the effect of chronic treatment with benzodiazepines on several parameters of hippocampal glutamatergic neurotransmission. METHODS: Rats were injected (IP) with a single dose or daily doses (21 days) of 1 mg/kg lorazepam (LZ) or vehicle. Thirty minutes after the last dose, animals were killed and parameters were measured in the dissected hippocampi. We determined one presynaptic parameter, in vitro glutamate release induced by a 60 mM K(+) stimulus. [(3)H]MK-801 binding to postsynaptic NMDA receptors and the NMDA stimulated efflux of cGMP were also evaluated. RESULTS: While no changes were observed in any of the parameters after a single dose of the drug, we found an increase of 206% in in vitro glutamate release in chronically treated animals [two-way ANOVA: F(1,16)=6.22], together with an increment of 103% in the NMDA stimulated cGMP efflux [two-way ANOVA: F(1,18)=14.05]. No changes either in K(D) or in B(max) values for [(3)H]MK-801 binding to hippocampal membranes were observed. CONCLUSIONS: Taken together, these changes strongly suggest that a compensatory increase in the glutamatergic response develops in the hippocampus during chronic treatment with LZ. Our findings might indicate a contribution of glutamatergic mechanisms to the tolerance to hippocampal-mediated effects of LZ, such as amnesic and anticonvulsant activities. PMID- 12545334 TI - Hepatic amino-acid metabolism in liver cirrhosis and in the long-term course after liver transplantation. AB - The aim of this study was to investigate the impact of orthotopic liver transplantation (OLT) on plasma levels and splanchnic turnover of key amino acids for muscular (branched-chain amino acids: BCAAs) and hepatic metabolism (aromatic amino acids (AAAs) and methionine) in 48 patients with cirrhosis, 14 patients after OLT, and 46 controls. Also, hepatic amino-acid supply and resting energy expenditure were measured. BCAA levels (no hepatic uptake) decreased in cirrhosis (P<0.001) and were improved, although not normalized, after OLT (P<0.001). AAA and methionine levels were raised in cirrhosis (P<0.001) and normalized after OLT (P<0.001). Hepatic supply of these amino acids increased in patients graded Child B and C and decreased significantly after OLT. Splanchnic uptake of AAAs and methionine increased significantly in Child-B and decreased in Child-C patients. After OLT, splanchnic extraction of AAAs and methionine was as in Child A. Circulating AAAs and methionine correlated with indocyanine-green half-life (r=0.71, P<0.001) and resting energy expenditure (r=0.50, P<0.001), indicating that levels of circulating AAAs and methionine in cirrhosis are determined by hepatic and extra-hepatic metabolic factors. This study demonstrates persistent changes in muscular metabolism of BCAAs after OLT, while the hepatic amino-acid metabolism is normalized due to (1) a significant reduction in the rate of peripheral proteolysis, and (2) improved liver function compared with that in patients with cirrhosis. PMID- 12545335 TI - Differential intragraft cytokine messenger RNA profiles during rejection and repair of clinical heart transplants. A longitudinal study. AB - After clinical heart transplantation, ischemia, acute rejection, and repair mechanisms can trigger the up-regulation of cytokines. To investigate the cytokine profile early after transplantation, we monitored messenger RNA (mRNA) expression levels of tumor necrosis factor-alpha (TNF-alpha), monocyte chemoattractant protein-1 (MCP-1), transforming growth factor-beta (TGF-beta), platelet-derived growth factor-A (PDGF-A), and basic fibroblast growth factor (bFGF) by reverse transcriptase-polymerase chain reaction (RT-PCR) in serial endomyocardial biopsies ( n=123) from 16 cardiac allograft recipients during the first 3 post-operative months. In the first month, mRNA expression levels of MCP 1, TNF-alpha, TGF-beta, and bFGF were significantly higher than in the period thereafter (acute rejection episodes excluded). Acute rejection (International Society for Heart and Lung Transplantation (ISHLT) rejection grade >2) was strongly associated with the level of TNF-alpha mRNA. After acute rejection episodes, rising mRNA expression levels of PDGF-A and bFGF were found. The association between TNF-alpha mRNA and acute rejection reflects the importance of this cytokine in allogeneic responses. Elevated growth factor expression levels indicate repair responses after tissue damage due to either the transplantation procedure (surgery, ischemia, reperfusion) or acute allograft rejection. PMID- 12545336 TI - Outcome of patients with pre-existing portal vein thrombosis undergoing arterialization of the portal vein during liver transplantation. AB - Arterialization of the portal vein is being propagated as a technical possibility in liver transplant recipients with pre-existing portal vein thrombosis. In our own small series, portal vein arterialization (PVA) was carried out in four patients undergoing orthotopic liver transplantation. In three of these cases, the portal vein was anastomosed to the aorta via an interposed iliac artery, and in one case, directly to the hepatic artery. After PVA, all transplants showed regular initial function. Two patients died postoperatively after 19 and 50 days, of intra-abdominal haemorrhage and liver necrosis with thrombosis of the portal vein, respectively. A further patient had previously developed fibrosis of the liver, which led to the death of the patient 11 months after PVA. In the remaining patient, chronic rejection requiring re-transplantation developed 24 months after PVA had been performed. These unfavourable results prompt the conclusion that PVA cannot be recommended as a standard clinical procedure. PMID- 12545337 TI - Improving donor identification with the Donor Action programme. AB - The increasing demand for organs for transplantation entails a consensual need for enhancement of organ procurement activity. As organ donors reside mainly in hospital intensive care units (ICUs), the Donor Action programme is aimed at identifying critical areas in ICUs, in order to improve the first step of organ donation. The purpose of this paper is to analyse the problem of identification of potential donors by means of a chart revision of patients who died in 14 ICUs in the Emilia-Romagna region between 1 July 1998 and 31 December 2000. All deaths and patients with severe brain insult (score on Glasgow Coma Scale (GCS) = 3/15) were assessed by the local transplant coordinators together with a professional at the Transplant Reference Centre. Brain death diagnoses and potential donor referrals were therefore assessed in the study period, which was subdivided into five semesters. Of the 3,056 deaths reported in 30 months, 1,248 were due to severe brain damage (GCS score = 3). Brain death diagnosis (BD) was performed in 509 patients (40.8%). Although we applied the same parameters over the whole length of the study, we observed a significant increase in BDs (from 87 in the first semester to 125 in the last, 30.5% to 53.0% of the patients with GCS 3 ( P=0.003, chi(2) for trend=16.072), in spite of a slight decrease in the total number of deaths and in the total number of patients with GCS score = 3 (from 649 to 587, and from 44% to 41%, respectively). Study population characteristics could have contrasted with rather than facilitated our results: age and gender did not change significantly, whilst cause of death showed a significant reduction in trauma and an increase in cerebrovascular incidents over the whole length of the study. We can conclude that the more careful assessment of patients dying in ICUs, by the Donor Action programme, significantly contributed to the improvement of BDs observed in the study period. Therefore, Donor Action seems to be an efficient quality control programme to improve identification of potential donors, the first stage of organ procurement. PMID- 12545338 TI - Two good reasons for an angiotensin-II type 1 receptor blockade with losartan after cardiac transplantation: reduction of incidence and severity of transplant vasculopathy. AB - Despite considerable progress in immunosuppressive therapy, the incidence and severity of transplant vasculopathy (TVP) after cardiac transplantation have not declined. The renin-angiotensin system (RAS) plays a pivotal role in the proliferation of vascular smooth muscle cells (VSMCs) contributing to TVP. We compared the effects of an angiotensin-II blocker, losartan (AT(1) blocker), and an angiotensin-converting enzyme (ACE) inhibitor, enalapril, on the incidence of diseased vessels and the severity of experimental TVP in the Lewis-to-Fischer rat heterotopic heart transplantation model. Recipients were randomly divided into six groups, group 1: no therapy, group 2: 3 mg/kg per day cyclosporine (CyA) s.c., group 3: CyA and 10 mg/kg per day losartan p.o., group 4: CyA and 40 mg/kg per day enalapril p.o., and groups 5 and 6: as groups 3 and 4, but additionally pre-treated with losartan or enalapril 7 days prior to transplantation. Eighty days after grafting, we assessed the incidence and severity of TVP, expressed as percentage of diseased vessels and mean vessel occlusion (MVO), by digitizing morphometry. CyA and CyA/enalapril post-treatment significantly reduced MVO, compared with controls, but not the incidence. Additional reduction of MVO was achieved in CyA/enalapril pre-treatment and both CyA/losartan pre- and post treatment groups when compared with CyA and untreated controls. However, only losartan post-treatment in combination with CyA reduced both incidence and MVO. Our results validate the important role of the RAS in neointimal proliferation after cardiac transplantation. Losartan appears to be superior to enalapril in preventing TVP after experimental cardiac transplantation. Therefore, AT(1) blockade with losartan might be a therapeutic option for the prevention of TVP in human heart recipients. PMID- 12545339 TI - Increased mortality after liver transplantation for hepatocellular carcinoma in hepatitis B-associated cirrhosis. AB - Transplant patients suffering from hepatocellular carcinoma in cirrhosis are selected according to tumor nodule number and diameter. Vascular invasion and histopathological grading are predictive of outcome. The prognostic influence of hepatitis B-cirrhosis has been investigated after resection and after local tumor treatment, but not after transplantation. Of the 1,188 transplantations performed between 1989 and 2000, 120 were on patients with hepatocellular carcinoma in cirrhosis (HCC) (follow-up: 57 months; 1-140 months). Within this group, 25 patients (21%) suffered from hepatitis B. Pre-transplant selection criteria were a maximum diameter of 5 degrees cm in uni-nodular tumors, or 3 degrees cm for two to three tumor nodules. The rate of tumors with 2-3 tumor nodules was increased in the hepatitis-B group (52% vs. 29%; P<0.05). Other tumor characteristics did not differ. In the hepatitis-B group, more patients died post-transplantation (44% vs.22%; P<0.05). This difference was due to unspecific causes, not to tumor recurrence or re-infection. These findings may be indicative of a more complicated course in patients suffering from hepatitis B in general. PMID- 12545340 TI - Busulfan depletes neutrophils and delays accelerated acute rejection of discordant xenografts in the guinea pig-to-rat model. AB - Complement factor C6 plays a critical role in mediating hyperacute rejection of discordant xenografts. In order to explore the mechanism of discordant xenograft rejection, we investigated kinetics and phenotypes of the cellular infiltrate in xenografts in untreated and leukocyte-depleted recipients, in relation to graft survival. Guinea pig cardiac xenografts were heterotopically transplanted to totally C6-deficient PVG (C-) rats. Grafts were removed after 0, 6, 12, and 24 h ( n = 6). Histological evaluation was performed with hematoxylin and eosin (H & E) and immunoperoxidase staining. The agents fucoidin and busulfan were applied to delay xenograft rejection further. Within 6 h, minimal perivascular edema with isolated infiltrating CD11b/c- and ED1-positive cells were found. An intense infiltration of CD11b/c- and ED1-positive cells with interstitial hemorrhage was present after 24 h, though with little CD161 and CD3 cell infiltration. Inhibition of cell adhesion by fucoidin did not prolong xenograft survival (34 +/ 15 h, n = 4, P<0.47), but the depletion of granulocytes by injection of busulfan did prolong survival of the discordant xenografts, to 62 +/- 22 h ( n = 7, P < or = 0.0039). These results demonstrate a significant effect of specific depletion of granulocytes and macrophages by busulfan therapy on guinea pig cardiac xenograft survival in PVG (C-) rats, suggesting the participation of these infiltrating cells in the xenoreactive rejection process. PMID- 12545341 TI - Safety and efficacy of an alternative basiliximab (Simulect) regimen after renal transplantation: administration of a single 40-mg dose on the first postoperative day in patients receiving triple therapy with azathioprine. AB - This was a multi-center, open-label, randomized, dose-comparative study on 202 renal transplantation patients. We evaluated for the first time an alternative dosing regimen for basiliximab, consisting of a single 40-mg intravenous dose on day 1 post-transplantation plus triple therapy, in comparison with the conventional two-dose regimen (2 h before transplantation and on day 4) plus triple therapy. At 6 months, the incidence of acute rejection was low: 22.5% of patients in the basiliximab 2 x 20-mg group and 20.0% of patients in the basiliximab 1 x 40-mg group experienced an acute rejection episode ( P = 0.628) (biopsy-proven rejection: 19.6% and 17.0%, P = 0.585). There was no statistically significant difference in any of the secondary efficacy parameters. The incidence of graft loss by 12 months was 4.9% and 6.0% in the 2 x 20-mg and 1 x 40-mg group, respectively ( P = 0.73). No differences were observed between the dosage groups with regards to safety assessments (adverse events (AEs), infections, vital signs, laboratory safety evaluations, and physical examinations). The data reveal that basiliximab can be safely and effectively administered as a single 40 mg dose on day 1 after renal transplantation as a therapeutic option to the established 2 x 20-mg dosing regimen. This alternative dosing regimen may be of significant convenience under circumstances when a first dose of basiliximab was not given prior to transplantation. Both regimens can conveniently be used during the initial hospitalization of the patient. PMID- 12545342 TI - Fluvastin therapy affects TAFI concentration in kidney transplant recipients. AB - Thrombin activatable fibrinolysis inhibitor (TAFI) is a glycoprotein, linking coagulation and fibrinolysis. Recently, attention has been drawn to the beneficial effects of statins on haemostasis in kidney patients prone to dyslipidaemia and with a high risk of cardiovascular death. The purpose of this study was to assess whether fluvastatin affects TAFI concentration in renal transplant recipients. We evaluated thrombin-antithrombin (TAT) complexes, prothrombin fragments 1+2, thrombomodulin, plasmin-antiplasmin (PAP) complexes, TAFI, P-selectin, and lipoprotein (a), 1, 2, and 3 months before and after fluvastatin treatment and in normolipaemic kidney transplant recipients and healthy volunteers. Cholesterol and LDL fell significantly as soon as 1 month after treatment had begun and remained lowered during the therapy. TAFI and prothrombin fragments 1+2 decreased significantly after 3 months of fluvastatin administration, whereas P-selectin decreased significantly after 2 months and remained significantly lower after 3 months of this therapy. We can conclude that fluvastatin is an effective hypolipaemic agent that favourably affects haemostasis. PMID- 12545343 TI - A new and objective definition of the term "indefinite survival" in organ transplantation in the animal model. PMID- 12545344 TI - Portable computed tomography performed on the intensive care unit. AB - OBJECTIVE: We report on the use of portable computed tomography (CT) in an ICU setting. The additional diagnostic gain and therapeutic consequences were assessed. PATIENTS: Ten ICU patients underwent 14 portable chest CT examinations. In 64% maximum intensive care was required, according to TISS28 (>40), and 42% were at a risk of mortality higher than 25% (MODS) on the day of portable CT examination. In three portable CT examinations the patients were considered not transportable and were examined directly in the patient room. All other examinations were performed in a special interventional suite directly on the ICU. RESULTS: Of 14 examinations 8 (57%) resulted in a change in patient management within 48 h. All patients profited from portable CT and no hazards occurred related to CT. CONCLUSIONS: To perform portable CT in the interventional suite on the ICU allows immediate minimally invasive therapeutic interventions and provides full ICU monitoring. PMID- 12545345 TI - Heart enlargement after thrombolysis for unsuccessful resuscitation. AB - Thrombolysis has been recently reported for the treatment of unsuccessful out-of hospital cardiopulmonary resuscitation. This case report presents the first case of mediastinal hematoma following thrombolysis in this setting. Chest X-ray and hemodynamic pattern mimicked pericardial effusion with cardiac-filling compromise. Diagnosis was established by transesophageal echocardiography and chest computerized tomodensitometry. PMID- 12545346 TI - Polychlorinated dibenzo-p-dioxins and dibenzofurans in breast milk from Chinese schistosomiasis areas. PMID- 12545347 TI - Levels of organochlorine pesticides in Beijing human milk, 1998. PMID- 12545348 TI - Dioxins/furans and polychlorinated biphenyls (PCBs) in Dugongs from the Thailand Coast. PMID- 12545349 TI - Polycyclic aromatic hydrocarbons (PAHs) and aliphatic hydrocarbons (AHs) in edible fish from the Arabian Gulf. PMID- 12545350 TI - Reduction of pesticide residues of fruit using water only or Plus Fit Fruit and Vegetable Wash. PMID- 12545351 TI - Evaluation of methyltin and butyltin pollution in Beijing Guanting reservoir and its downriver Yongding river. PMID- 12545352 TI - Validity test for a yeast two-hybrid assay to screen for estrogenic activity, and its application to insecticides and disinfectants for veterinary use. PMID- 12545353 TI - Copper metallothionein-like proteins as exposure biomarker in native and transplanted intertidal populations of the mussel Perumytilus purpuratus from San Jorge Bay, Antofagasta, Chile. PMID- 12545354 TI - Marine benthic microalgae Cylindrotheca closterium (Ehremberg) Lewin and Reimann (Bacillariophyceae) as a tool for measuring toxicity of linear alkylbenzene sulfonate in sediments. PMID- 12545355 TI - Degradation of nonylphenol ethoxylates (NPE) in sewage sludge and source separated municipal solid waste under bench-scale composting conditions. PMID- 12545356 TI - Biodegradation of phthalic acid esters by bakery yeast Saccharomyces cerevisiae. PMID- 12545358 TI - Biotreatment of distillery effluent using Aspergillus niveus. PMID- 12545359 TI - Effects of moult, age, and sex on the accumulation of heavy metals in the otter (Lutra lutra) in Finland. PMID- 12545357 TI - Removal of zinc and fluoride ions from industrial waste water plants around Cairo. PMID- 12545360 TI - Reproductive toxicity of endosulfan in male albino rats. PMID- 12545361 TI - Effect assessment of antimicrobial pharmaceuticals on the aquatic plant Lemna minor. PMID- 12545362 TI - Acute toxicity of the synthetic pyrethroid deltamethrin to freshwater catfish Clarias gariepinus. PMID- 12545363 TI - Cytogenetic effect of organoarsenicals on cultured mammalian cells. PMID- 12545364 TI - Australian freshwater crayfish Cherax destructor accumulates and depurates nickel. PMID- 12545365 TI - Effects of 4-nonylphenol on sexual maturation in Daphnia magna. PMID- 12545366 TI - Chromosomal aberrations in the leucocytes of men occupationally exposed to uranyl compounds. PMID- 12545367 TI - Isocyanates in flexible polyurethane foams. PMID- 12545368 TI - Human exposure assessment to mancozeb during treatment of mandarin fields. PMID- 12545369 TI - Comparison of active and passive measurement of ozone in Zagreb air. PMID- 12545370 TI - Organochlorine pesticide residues in water from Guanting Reservoir and Yongding River, China. PMID- 12545371 TI - Fecal excretion of dioxin in mice enhanced by intake of dietary fiber bearing chlorophyllin. PMID- 12545372 TI - Preliminary study of an enzyme extracted from Alcaligenes sp. strain YF11 capable of degrading pesticides. PMID- 12545373 TI - Sediment quality of Singanalluar Wetland in Coimbatore, Tamil Nadu, India. PMID- 12545374 TI - Mercury in river water and sediments in some rivers near Dunkwa-On-Offin, an Alluvial Goldmine, Ghana. PMID- 12545375 TI - Concentrations of metals in feathers and blood of nestling Black-crowned Night Herons (Nycticorax nycticorax) in Chesapeake and Delaware Bays. PMID- 12545376 TI - Biliary PAH metabolite elimination in Australian bass, Macquaria novemaculeata following exposure to bass strait crude oil and chemically dispersed crude oil. PMID- 12545377 TI - Stress-related bioindicator anomalies in feral male winter flounder (Pleuronectes americanus) exposed to effluent from two pulp and paper mills in Newfoundland. PMID- 12545378 TI - Acute and sublethal toxicity of thallium to aquatic organisms. PMID- 12545379 TI - Efficient mechanical disruption of Lactobacillus helveticus, Lactococcus lactis and Propionibacterium freudenreichii by a new high-pressure homogenizer and recovery of intracellular aminotransferase activity. AB - Microbiological studies often involve bacterial cell fractionation, which is known to be difficult for Gram-positive as compared to Gram-negative bacteria. Our purpose was to test the breaking efficiency of a new high-pressure pilot homogenizer for three Gram-positive species involved in dairy technology and to assess the activity of an intracellular aminotransferase. Varied pressures (50, 100 and 200 MPa) were applied to concentrated bacterial suspensions (1.2 mg dry weight/ml) of Lactobacillus helveticus, Lactococcus lactis and Propionibacterium freudenreichii. Breaking efficiency was estimated by decreases in optical density at 650 nm, cellular dry weight and viability. The proteins released were quantified and the residual intracellular aminotransferase activity was estimated using leucine as substrate. One run at 50 MPa was sufficient to break 80% of lactobacilli cells whereas 200 MPa were required for the same efficiency for L. lactis and P. freudenreichii. Whatever the pressure, leucine aminotransferase activity was recovered in the supernatant after cell breaking. This new high pressure pilot homogenizer can allow rapid (20 s/run), easy, continuous and highly efficient cell breaking for intracellular enzyme recovery or other purposes. As the species tested were not phylogenetically related, and had different morphologies and cell wall compositions, we conclude that most Gram positive bacteria may be broken efficiently by this new device. PMID- 12545380 TI - Recombinant carbazole-degrading strains for enhanced petroleum processing. AB - Biotechnological upgrading of fossil fuels is of increasing interest as remaining stocks of petroleum show increasing levels of contaminants such as heavy metals, sulfur and nitrogen-containing heteroaromatic compounds. Carbazole is of particular interest as a major petroleum component known to reduce refining yields through catalyst poisoning. In this study, the biotransformation of carbazole was successfully demonstrated in a liquid two-phase system, when solubilized in either 1-methylnaphthalene or in diesel fuel. The effects of solvent toxicity were investigated by expressing the carbazole-transformation genes from MB1332, a rifampicin-resistant derivative of Pseudomonas sp. LD2, in a solvent-resistant heterologous host, P. putida Idaho [1]. This solvent-resistant strain successfully degraded carbazole solubilized in 1-methylnaphthalene and in the presence of 10 vol% xylenes similar to the non-recombinant strain Pseudomonas sp. LD2. Identification of a suitable recombinant host, however, was essential for further investigations of partial pathway transformations. Recombinant P. putida Idaho expressing only the initial dioxygenase enzymes transformed carbazole to an intermediate well retained in the oil phase. Partial carbazole transformation converts carbazole to non-aromatic species; their effect is unknown on refinery catalyst poisoning, but would allow almost complete retention of carbon content and fuel value. PMID- 12545382 TI - Lactic acid production by Saccharomyces cerevisiae expressing a Rhizopus oryzae lactate dehydrogenase gene. AB - This work demonstrates the first example of a fungal lactate dehydrogenase (LDH) expressed in yeast. A L(+)-LDH gene, ldhA, from the filamentous fungus Rhizopus oryzae was modified to be expressed under control of the Saccharomyces cerevisiae adh1 promoter and terminator and then placed in a 2 micro -containing yeast replicating plasmid. The resulting construct, pLdhA68X, was transformed and tested by fermentation analyses in haploid and diploid yeast containing similar genetic backgrounds. Both recombinant strains utilized 92 g glucose/l in approximately 30 h. The diploid isolate accumulated approximately 40% more lactic acid with a final concentration of 38 g lactic acid/l and a yield of 0.44 g lactic acid/g glucose. The optimal pH for lactic acid production by the diploid strain was pH 5. LDH activity in this strain remained relatively constant at 1.5 units/mg protein throughout the fermentation. The majority of carbon was still diverted to the ethanol fermentation pathway, as indicated by ethanol yields between 0.25-0.33 g/g glucose. S. cerevisiae mutants impaired in ethanol production were transformed with pLdhA68X in an attempt to increase the lactic acid yield by minimizing the conversion of pyruvate to ethanol. Mutants with diminished pyruvate decarboxylase activity and mutants with disrupted alcohol dehydrogenase activity did result in transformants with diminished ethanol production. However, the efficiency of lactic acid production also decreased. PMID- 12545381 TI - Silicone antifoam performance enhancement by nonionic surfactants in potato medium. AB - The ability of a silicone antifoam to retard foaming in a liquor prepared from potatoes is enhanced by the addition of ethoxylated nonionic surfactants. The enhancement is non-linear for surfactant concentration, with all 12 surfactants tested possessing a concentration at which foam heights strongly diminish, referred to as the surfactant critical antifoaming concentration (SCAFC). SCAFCs vary between surfactants, with lower values indicating better mass efficiency of antifoaming enhancement. SCAFCs decrease with degree of ethoxylation and decrease with the hydrophilic-lipophilic balance for ethoxylated nonionic surfactants. Surfactant addition produces a mixed water-surface layer containing surfactant and surface-active components in the potato medium. Surface tension reduction does not correlate well with antifoam performance enhancement. A model is proposed where surfactant adsorption promotes desorption of surface-active potato medium components from the water surface. At the SCAFC, desorption is not complete, yet the rate of bubble rupture is sufficiently enhanced to provide excellent foam control. PMID- 12545383 TI - Effect of culture conditions on manganese peroxidase production and activity by some white rot fungi. AB - The ligninolytic system of white rot fungi is primarily composed of lignin peroxidase, manganese peroxidase (MnP) and laccase. The present work was carried out to determine the best culture conditions for production of MnP and its activity in the relatively little-explored cultures of Dichomitus squalens, Irpex flavus and Polyporus sanguineus, as compared with conditions for Phanerochaete chrysosporium and Coriolus versicolor. Studies on enzyme production under different nutritional conditions revealed veratryl alcohol, guaiacol, Reax 80 and Polyfon H to be excellent MnP inducers. PMID- 12545384 TI - The effect of carbon sources and lactate dehydrogenase deletion on 1,2 propanediol production in Escherichia coli. AB - In previous studies, we showed that cofactor manipulations can potentially be used as a tool in metabolic engineering. In this study, sugars similar to glucose, that can feed into glycolysis and pyruvate production, but with different oxidation states, were used as substrates. This provided a simple way of testing the effect of manipulating the NADH/NAD+ ratio or the availability of NADH on the metabolic patterns of Escherichia coli under anaerobic conditions and on the production of 1,2-propanediol (1,2-PD), which requires NADH for its synthesis. Production of 1,2-PD was achieved by overexpressing the two enzymes methylglyoxal synthase from Clostridium acetobutylicum and glycerol dehydrogenase from E. coli. In addition, the effect of eliminating a pathway competing for NADH by using a ldh(-) strain (without lactate dehydrogenase activity) on the production of 1,2-PD was investigated. The oxidation state of the carbon source significantly affected the yield of metabolites, such as ethanol, acetate and lactate. However, feeding a more reduced carbon source did not increase the yield of 1,2-PD. The production of 1,2-PD with glucose as the carbon source was improved by the incorporation of a ldh(-) mutation. The results of these experiments indicate that our current 1,2-PD production system is not limited by NADH, but rather by the pathways following the formation of methylglyoxal. PMID- 12545385 TI - Production of the secondary metabolites gamma-aminobutyric acid and monacolin K by Monascus. AB - gamma-Aminobutyric acid (GABA), a hypotensive agent, and monacolin K, a cholesterol-lowering drug, can be produced by Monascus spp. Under optimal culture conditions, the products of fermentation using Monascus spp. may serve as a multi functional dietary supplement and can prevent heart disease. In this study, Monascus purpureus CCRC 31615, the strain with the highest amount of monacolin K, was identified from 16 strains using solid fermentation. Its GABA productivity was particularly high. Addition of sodium nitrate during solid-state fermentation of M. purpureus CCRC 31615 improved the productivity of monacolin K and GABA to 378 mg/kg and 1,267.6 mg/kg, respectively. GABA productivity increased further to 1,493.6 mg/kg when dipotassium hydrophosphate was added to the medium. PMID- 12545386 TI - Immobilisation of cyclodextrin glucanotransferase from Bacillus circulans ATCC 21783 on purified seasand. AB - Cyclodextrin glucanotransferase (CGTase) from Bacillus circulans (ATCC 21783) was immobilised on a silica-based support: purified seasand. Although adsorption of 98% was achieved, considerable desorption was encountered. This problem was minimised by crosslinking the adsorbed enzyme with glutaraldehyde. The immobilised enzyme after crosslinking could be used repeatedly for cyclodextrin (CD) production in a batch process. The activity retention was 80% at the end of the eighth cycle. The immobilised enzyme showed a shift in the pH optimum towards the alkaline side and also an improvement in the pH stability compared to the free enzyme. It catalysed the formation of beta-CD as a major product. A significant amount of alpha-CD production was also observed on prolonged incubation. PMID- 12545387 TI - Evaluation of Kluyveromyces marxianus as a source of yeast autolysates. AB - Cells of Kluyveromyces marxianus FII 510700 and Saccharomyces cerevisiae CBS 1907 were autolysed in phosphate buffer, pH 4.5, for a maximum of 10 days to compare chemical changes that occur in the carbohydrate, protein, amino acid and nucleic acid content. Approximately 2.2-3% carbohydrate, 9.5-12% protein, 0.6-1.0% DNA and 6-7% RNA were recovered in the autolysates. The main amino acids were beta alanine, phenylalanine, cysteine, methionine, glutamic acid and isoleucine. No significant differences in the yeast autolysates of K. marxianus and S. cerevisiae were observed. Consequently, K. marxianus produced from lactose-based media has potential as a source of yeast autolysates used in the food industry. PMID- 12545388 TI - Microarray analyses of the metabolic responses of Saccharomyces cerevisiae to organic solvent dimethyl sulfoxide. AB - The toxic effects that organic solvents have on whole cells are important drawbacks in the application of these solvents in the production of fine chemicals by whole-cell stereoselective biotransformations. Although early studies found that organic solvents mainly destroyed the integrity of cell membranes by accumulating in the lipid bilayer of plasma membranes, the cellular metabolic responses to the presence of an organic solvent remain unclear. With the rapid development of genomics, it is possible to study cellular metabolism under perturbed conditions at the genome level. In this paper, the global gene expression profiles of Saccharomyces cerevisiae BY4743 grown in media with a high concentration of the organic solvent dimethyl sulfoxide (DMSO) were determined by microarray analysis of ~6,200 yeast open reading frames (ORFs). From cells grown in SD minimal medium containing 1.0% (v/v) DMSO, changes in transcript abundance greater than or equal to 2.5-fold were classified. Genomic analyses showed that 1,338 genes were significantly regulated by the presence of DMSO in yeast. Among them, only 400 genes were previously found to be responsive to general environmental stresses, such as temperature shock, amino acid starvation, nitrogen source depletion, and progression into stationary phase. The DMSO responsive genes were involved in a variety of cellular functions, including carbohydrate, amino acid and lipid metabolism, cellular stress responses, and energy metabolism. Most of the genes in the lipid biosynthetic pathways were down regulated by DMSO treatment, whereas genes involved in amino acid biosynthesis were mostly up-regulated. The results demonstrate that the application of microarray technology allows better interpretation of metabolic responses, and the information obtained will be useful for the construction of engineered yeast strains with better tolerance of organic solvents. PMID- 12545389 TI - Degradation of polyvinyl alcohol by Sphingomonas sp. SA3 and its symbiote. AB - A total of 800 samples was taken from Taegu province, Korea, where many textile factories provide a source of polyvinyl alcohol (PVA) waste. These samples were screened for PVA-degrading bacteria. A new strain, SA3, was discovered which formed yellow colonies and used PVA as the sole carbon and energy source. Strain SA3 was identified as a Sphingomonas sp., based on the partial nucleotide sequence analysis of 16S ribosomal RNA, the presence of 2-hydroxymyristic acid (14:O 2-OH) and sphingolipids with d-17:0, d-18:0, d-19:1, and d-20:1 as the main dihydrosphingosines. This genus has not previously been reported as a PVA degrading bacterium. Sphingomonas sp. SA3 needs a symbiote strain, SA2, for PVA degradation as a growth factor producer. In mixed cultures of these strains, the optimum temperature for PVA biodegradation ranged from 30 degrees C to 35 degrees C. The optimum pH was 8.0 and the most effective nitrogen source was NH(4)(+). PMID- 12545390 TI - An inexpensive medium for production of arachidonic acid by Mortierella alpina. AB - The production of arachidonic acid was studied in the fungus Mortierella alpina using an inexpensive medium. Glucose derived from maize starch hydrolysate was the sole carbon source and defatted soybean meal and sodium nitrate were the nitrogen sources. Optimal arachidonic acid yield (1.47 g l(-1)) was observed at a glucose concentration of 100 g l(-1). Various treatments of defatted soybean meal to extract soluble nitrogen nutrients were evaluated. Alkali extract was the most effective for arachidonic acid production. A mixture of soybean alkali-extract protein and sodium nitrate was an excellent nitrogen source for fungal growth, lipid accumulation, and arachidonic acid production. A maximum yield of 1.87 g arachidonic acid l(-1) was obtained with a soybean protein concentration of 4.6 g l(-1) and a sodium nitrate concentration of 2.3 g l(-1). PMID- 12545406 TI - Cerebral malignant nerve sheath tumor, triton tumor variant: case report. AB - A case of a cerebral malignant triton tumor in a 3-year-old boy with a 2-month history of frontal headache and no clinical evidence of neurofibromatosis is reported. The computed tomography (CT) scan showed a large, irregular tumor in the right parietooccipital lobe. A partial surgical resection was performed. Histologically, the tumor was highly cellular and consisted of spindle cells with hyperchromatic and pleomorphic nuclei. Focally, neoplastic cells with rhabdomyoblastic features were found. The immunohistochemical study showed that tumor cells were positive for S-100 protein and CD57, and the rhabdomyoblasts expressed desmin, Myo-D1, and myoglobin. During the postoperative period, a massive intraparenchymal hemorrhage was identified and surgical drainage was performed. The patient worsened and died 10 days after the first surgery. Postmortem study was not authorized. Six cases of cerebral malignant nerve sheath tumor have been described; however, primary intraparenchymal malignant triton tumor has not been previously described. PMID- 12545405 TI - Soft tissue perineuriomas in children: report of three cases and review of the literature [corrected]. AB - Perineuriomas (PN) are uncommon, slowly growing, usually benign tumors composed of well-differentiated perineural cells. Two variants are recognized: intraneural perineuriomas and soft tissue perineurioma, which includes a sclerosing subset of tumors. They are usually reported in the adult population. We present three cases of soft tissue perineuriomas in children. One was located in the deep soft tissue of the retroperitoneum in a 14-year-old girl, the second one in the left thumb of a 14-year-old boy, and the third one in the index finger of a 16-year-old boy. This report, which describes the clinicopathologic, immunohistochemical, and ultrastructural features of these tumors, should alert pathologists to the occurrence of perineuriomas in children. A review of the English language literature on perineuriomas in children is also included. PMID- 12545407 TI - Sexual diversity in urban Norwegians. AB - The purpose of this study is to describe homosexual and heterosexual experiences in terms of sexual fantasies, sexual attraction, sexual conduct, and falling in love in an urban Norwegian population. In 1997, a random sample of 5,000 persons (18-49 years) was drawn from the population register in Oslo to participate in a sexual behavior survey. Data collection was carried out by means of postal questionnaires and 45% responded. The results indicated that exclusive homosexuality was rare in the population. One exception was the prevalence of homosexual fantasies in women. Compared to respondents who had had exclusively heterosexual contact, respondents reporting bisexual contacts had a significantly higher number of lifetime sex partners, higher frequency of anal and oral sex and masturbation, and lower age at orgasm and masturbation debut. What significantly separated the groups of exclusive heterosexuals, bisexuals, and exclusive homosexuals were attitudes towards various expressions of sexuality and number of lifetime sex partners. PMID- 12545408 TI - Reports of childhood sexual abuse by adolescents and young adults: stability over time. AB - The consistency of childhood sexual abuse (CSA) reporting was explored in this study. Two-hundred seventeen adolescents and young adults (ages 14-24) enrolled in urban health care clinics completed self-report questionnaires assessing CSA and other problem behaviors at enrollment and at 7 months. Results indicated that the stability of CSA self-report at two time points was poor (58% consistent nonreporters of CSA, 20% consistent reporters, 22% inconsistent reporters). Consistent and inconsistent reporters were differentiated on risk measures. Adolescents who endorsed more items from the CSA scale were five times more likely to be consistent reporters. In sum, adolescent CSA reporting was quite inconsistent over time. Using multi-item scales and assessing CSA at two time points enhances accuracy of reporting. PMID- 12545409 TI - Heterosexuals attitudes toward bisexual men and women in the United States. AB - This paper examines heterosexual adults attitudes toward bisexual men and women using data from a 1999 national RDD survey (N = 1,335). Ratings on 101-point feeling thermometers were lower (less favorable) for bisexual men and bisexual women than for all other groups assessed--including religious, racial, ethnic, and political groups--except injecting drug users. More negative attitudes toward bisexuals were associated with higher age, less education, lower annual income, residence in the South and rural areas, higher religiosity, political conservatism, traditional values concerning gender and sexual behavior, authoritarianism, and lack of contact with gay men or lesbians. White heterosexual women expressed significantly more favorable attitudes than other women and all men. A gender difference was observed in attitudes toward bisexuals and homosexuals: Heterosexual women rated bisexuals significantly less favorably than they rated homosexuals, regardless of gender, whereas heterosexual men rated male targets less favorably than female targets, regardless of whether the target was bisexual or homosexual. PMID- 12545410 TI - Predicting responses to sexually aggressive stories: the role of consent, interest in sexual aggression, and overall sexual interest. AB - Measures of interest in sexual aggression and overall sexual interest were used to predict men s responses to consensual and nonconsensual stories that contained forceful sexual content. College males (N = 108) listened to these stories and completed questionnaires assessing self-reported sexual arousal and desire, their perceptions of the woman s enjoyment and sexual desire in the stories, their likelihood of acting like the man in the stories, and how likely they would be to read similar stories. Multiple regression analyses revealed that overall sexual interest variables were significant predictors of responses to both consensual and nonconsensual stories, while interest in sexual aggression predicted responses to only the nonconsensual story. The present study highlights the importance of considering overall sexual interest measures in sexual aggression research. PMID- 12545411 TI - Relationship power, decision making, and sexual relations: an exploratory study with couples of Mexican origin. AB - This study explored how couples of Mexican origin define power in intimate relationships, what makes men and women feel powerful in relationships, and the role of each partner in decision making about sexual and reproductive matters. Interviews were conducted with each partner of 39 sexually active couples and data were analyzed using content analysis. Results indicate that power is perceived as control over one s partner and the ability to make decisions. Women say they feel more powerful in relationships when they make unilateral decisions and have economic independence. Men feel powerful when they have control over their partner and bring home money. Respondents agreed that women make decisions about household matters and children, while men make decisions related to money. Findings indicate that whereas couples share decision making about sexual activities and contraceptive use, men are seen as initiators of sexual activity and women are more likely to suggest condom use. PMID- 12545412 TI - The economy of risk and respect: accounts by Puerto Rican sex workers of HIV risk taking. AB - Three-hundred-eleven female drug-using sex workers in urban Puerto Rico were asked to describe their last negotiation with a client. They described efforts to protect themselves from many hazards of sex work, including violence, illness, and drug withdrawal. They also described efforts to minimize the stigma and marginalization of sex work by cultivating relationships with clients, distinguishing between types of clients, and prioritizing their role as mothers. Sex workers adopted alternating gender roles to leverage autonomy and respect from clients. Their narratives suggest that sex workers negotiate a world in which HIV is relative to other risks, and in which sexual practices which are incomprehensible from an HIV-prevention perspective are actually rooted in a local cultural logic. Future HIV prevention efforts should frame condom use and other self-protective acts in terms that build upon sex workers own strategies for understanding their options and modifying their risks. PMID- 12545413 TI - Relationship functioning and sexuality among people with multiple sclerosis. AB - This study was concerned with how the sexuality and relationships of people with Multiple Sclerosis (MS) are different from those of people from the general population. Three-hundred eighty-one respondents (144 males, 237 females) with MS and 291 respondents (101 males, 190 females) from the general population participated in the study. Sexual satisfaction, sexual dysfunction, relationship satisfaction, and coping style were assessed among all respondents. Information was also obtained from people with MS regarding age of onset and diagnosis of symptoms, as well as severity of symptoms. Overall, males with MS experienced a higher frequency of sexual dysfunction than males from the general population, while females with MS only differed from females from the general population in their levels of masturbation and numbness of the genital area. Coping strategies and levels of cognitive functioning were important predictors of sexual satisfaction, sexual dysfunction, and relationship satisfaction for women with MS, but there were fewer coping or health-related factors that predicted these variables among men with MS. The results of this study are discussed in terms of factors related to sexual satisfaction and positive interpersonal relationships. There is a need to conduct further research in this area so that professionals working with people with MS are informed on strategies to improve the sexuality and relationships of their clients lives. PMID- 12545414 TI - Understanding human ambivalence about sex: the effects of stripping sex of meaning. AB - We offer a theoretical perspective to provide insight into why people are ambivalent about sex and why cultures regulate sex and attach symbolic meaning to it. Building on terror management theory, we propose that sex is problematic for humankind in part because it reminds us of our creaturely mortal nature. Two experiments investigated the effects of reminding people of the similarity between humans and other animals on their reactions to the physical aspects of sex. In Study 1, priming human-animal similarities led to increased accessibility of death-related thoughts after thinking about the physical but not romantic aspects of sex. In Study 2, when participants were reminded of similarities between humans and other animals, mortality salience resulted in decreased attraction to the physical but not romantic aspects of sex. In each study, priming thoughts about how humans are distinct from animals eliminated the association between sex and death. PMID- 12545415 TI - Sexual functioning and self-reported depressive symptoms among college women. AB - We conducted an exploratory study comparing 47 college-aged women reporting depressive symptoms but not receiving antidepressant medication to 47 age-matched controls. We examined various dimensions of sexual functioning, including sexual desire, arousal, orgasm, pain, pleasure, and satisfaction. The women with depressive symptoms reported more inhibited sexual arousal, more inhibited orgasm, more sexual pain problems, and less sexual satisfaction and pleasure than control participants. Novel to this study, the women with depressive symptoms reported greater desire for sexual activity alone (masturbation) than the nondepressed women. The findings are discussed in terms of primary reinforcers and depressive symptomology. PMID- 12545416 TI - Peer modeling and college men's sexually impositional behavior in the laboratory. AB - The purpose of this study was to examine the influence of peer modeling on sexually impositional behavior in the laboratory. Male participants with and without a self-reported history of sexually aggressive behavior viewed video clips depicting nonaggressive and sexually aggressive behavior and then chose one of the clips to show to a female confederate. Half of the participants were first exposed to a male confederate who showed the sexually aggressive video clip to a female confederate. The other half of the participants were exposed to a male confederate who showed a nonaggressive video clip to a female confederate. Exposure to a male confederate who showed a sexually aggressive video clip to a female was associated with participants choosing to engage in this same behavior. A self-reported history of sexually aggressive behavior was also associated with participants showing the sexually aggressive video clip in spite of believing the effect on the female viewer would be negative. PMID- 12545418 TI - Influence of gestational age and intrauterine growth on leptin concentrations in venous cord blood of human newborns. AB - BACKGROUND: The ob gene product leptin is involved in the regulation of body weight and energy expenditure, suggesting a potential role of leptin in embryonal and fetal development and progression of pregnancy. In term infants, leptin concentrations showed a positive correlation with birth weight. We aimed at comparing leptin cord blood levels in AGA (appropriate for gestational age) to SGA (small for gestational age) preterm and term newborns. PATIENTS AND METHODS: Ninety-seven human newborns, 47 females and 50 males, 33 born at term and 64 born before 36 weeks of gestation, were studied prospectively. Leptin concentrations in venous cord blood were determined using a specific RIA (radioimmunoassay). RESULTS: In term newborns, mean gestational age (GA) was 39 weeks (wk) (+/- 0.7 wk) and mean birth weight (BW) was 3316 g (+/- 473 g); in preterm newborns (n = 64), mean GA was 30 wk (+/- 5.0 wk) and mean BW was 1398 g (+/- 505 g). Mean standard deviation score of birth weight (BW SDS) was calculated as - 0.47. Mean leptin concentrations in term newborns differed significantly from those in preterm newborns (9.21 +/- 2.63 ng/ml vs. 1.58 +/- 0.88 ng/ml; p < 0.0001). In preterm and term infants, leptin concentrations showed a linear correlation with BW (r = 0.46; p < 0.0001) and GA (r = 0.48; p < 0.0001), respectively. Leptin levels were best predicted by an exponential regression model with GA (Leptin = exp(- 4.41 + 0.14 x GA); r = 0.61; p < 0.0001). Using multivariate regression analysis (r = 0.57; p < 0.0001), we found significant influences of GA (p < 0.00001) and BW SDS (p < 0.05) on leptin levels. No difference was observed between leptin values in AGA versus SGA preterm infants. CONCLUSION: These data suggest fetal leptin levels to be primarily determined by GA and additionally modulated by growth restriction in term newborns. We found a dramatic increase at weeks 33 to 35 of gestation and no modulation by BW SDS in very preterm infants. PMID- 12545419 TI - [Lymphadenitis colli due to non-tuberculous mycobacteria (NTM): a case-series and review of the literature]. AB - OBJECTIVE: Lymphadenitis colli due to NTM should always be considered in children with cervical Lymphadenitis. For Germany there is a lack of data concerning the incidence, the epidemiology, the diversity and frequency of the different bacteria, the diagnosis, the clinical manifestation and the medical treatment. METHODS: By means of a questionnaire, which was retrospective for 1985 to 1994 and was sent to 277 children's hospitals in Germany, we collected data on Lymphadenitis colli in Germany. In our study we also incorporated cases from the "National Laboratory for Mycobacteria" in Borstel as well as six cases from our hospital in Mainz. Therefore our data includes both clinical (28) and laboratory (30) cases. Additionally we screened the literature on "Lymphadenitis colli in children due to NTM". RESULTS: A total of 51 cases of Lymphadenitis due to NTM could be identified. The illness occurs typically in young children up to six years of age. The most frequent cause were species of the Mycobacterium avium intracellulare-scrofulaceum complex. Except for the local diagnosis of a cervical Lymphadenitis other clinical symptoms are missing, just as specific laboratory parameters with a subacute or chronic course. The tuberculin skin test can be false positive. The diagnosis is confirmed by biopsy and histology as well as through microbiological tests. CONCLUSIONS: The best treatment is complete surgical excision, whereas the importance of additional or exclusive treatment with Clarithromycin, Rifabutin and other antibiotics could not be clarified completely. But in patients with AIDS Rifabutin and other drugs could perhaps be useful, even for prophylaxis. Also if complete excision is impossible, treatment with certain drugs (Clarithromycin or Azithromycin in combination with Rifampicin) will be recommended. It still remains in question if NTM infections in children are really increasing. PMID- 12545420 TI - [What is a Sufficient Blood Pressure in the Preterm Newborn?]. AB - Hypotension is diagnosed in a high percentage of very low birth weight infants, particularly in the first 24 hours after birth and the resultant clinical approach is to support the blood pressure vigorously. However, the research base is not yet available to give a definite answer to the question, which blood pressure is really unacceptable in a given clinical situation. The clinical approach usually relies on reference blood pressure data or on clinical considerations. Reference blood pressure ranges established from observations of rather stable preterm infants and commonly used in "normal" premature infants are age- and weight related so-called "normal" blood pressures and "rules of thumb" while cerebral autoregulation adapted or outcome related lower limits of arterial blood pressure could be promising alternatives. Clinical signs, results of laboratory studies and cardiac function monitoring are tools used in stable and unstable premature infants for assessing if an acute blood pressure is adequate to prevent tissue malperfusion. In this rather unsatisfactory situation there is a growing body of evidence that also in premature infants systemic or regional blood flow and not blood pressure are the crucial hemodynamic parameters and that also in this age group there is at best a weak correlation between mean arterial blood pressure and left ventricular output or regional blood flow. PMID- 12545421 TI - [Finger replantation in a 3-year-old child]. AB - In this case report, we present a three year old boy who suffered an amputation of his left index finger at the level of the nail base (proximal zone I) caused by a scissor cut. Following referral to a center with hand surgical and pediatric care, successful replantation was achieved and postoperative pediatric care ensured finger perfusion. Three months postop, the reconstructed finger was in normal daily use with full function. Our success encourages for initial replantation of digits in infants in any case and irrespective of the level of amputation. Optimal results in these patients can only be achieved by interdisciplinary care of hand surgeons and pediatricians. PMID- 12545422 TI - [Early and late complications after implantation of central venous catheters]. AB - The use of implantable central venous catheters by the puncture-technique of Nagy is a safe method performed by trained surgeons. Due to the high mobility and then reduction of painful blood samples this method contributes favourably to the improvement of the quality of life of children with chronic diseases. The analysis of 140 catheters implanted in the Department of Pediatric Surgery of the University of Leipzig between 1995 and 2000 showed 11 cases with early complications. As the most frequent late complications were infection and thrombosis in 51 children. Neutropenia is a particular risk factor during polychemotherapy of malignant tumors. Staphylococcus epidermidis was the most frequently isolated bacillus. Catheter associated infections are only partly treatable by antibiotic therapy. In 29 of 44 cases the explantation of the catheter was necessary. Only a strict hygienic regimen would minimize the risk of infection PMID- 12545423 TI - [Guillain-Barre syndrome as a rare cause of acute respiratory insufficiency]. AB - BACKGROUND: Guillain-Barre syndrome (GBS) is an acute inflammatory demyelinating disease of peripheral nerves. Antibodies acting against antigens on the myelin or the axons seem to play a causative role. In up to 80 % the onset of GBS follows an antecedent respiratory or gastrointestinal infection. CASE REPORT: An 20 month old boy was referred to our hospital because of meningism and aspiration pneumonia. 10 days earlier the patient had experienced a period of fever. Because of respiratory insufficiency the patient was intubated, sedated and received mechanical ventilation. That's why a major neurological examination was not possible. The patient demonstrated a flaccid tetraplegic paralysis and autonomic dysfunction with elevated blood pressure, tachycardia, elevated ADH level and hyperglycaemia. Decreased motor nerve conduction and an increased CSF protein with normal CSF cell count confirmed diagnosis of GBS. Active CMV infection was diagnosed by PCR as the possible trigger factor. Intravenous immunoglobulins were given and the patient reached a complete remission except a slight disturbance of peroneal nerve. CONCLUSIONS: IVIG and PP therapy are equally effective in GBS. In contrast corticosteroids are not of benefit. Prognosis of childhood GBS is good. Only 4 % of affected children demonstrate persistent muscular weakness. The long interval between admission and diagnosis in the reported case emphasized the importance of neurological examination especially in sedated patients. PMID- 12545424 TI - [Therapy of osteoid osteomas -- always surgically?]. AB - Osteoidosteomas are common bone tumours in childhood. Frequently they occur in the long bones of the lower extremities, less frequent in the humeri, phalanxes or the axial skeleton. The tumour is benign and noninfiltrative. Metastases do not occur. Typical complaints are nightly pain attacks, which are relieved by nonsteroidal antiinflammatory drugs. On X-ray, the classic finding is a small radiolucent area surrounded by sclerotic bone in the cortex. Surgical excision is often recommended, providing the possibility for a histological diagnosis. Therapeutic alternatives are percutaneous coagulation of the nidus by alcohol or laser, thermo-coagulation or high-frequency radioablation. Spontaneous remissions are well documented. Therefore the decision to wait and see and to treat with antiinflammatory medication is a considerable therapeutic option. We present two patient with an ostoidosteoma in the proximal femur. One of them underwent an unsuccessful trial for surgical extirpation and histological examination but afterwards developed a severe hip contraction with scoliosis. In the case of the other patient a primary operation was abandoned. Pain was completely controlled upon treatment with nonsteroidal antiinflammatory drugs in both patients. The contractions also disappeared completely upon physiotherapy. Serial MRI showed a significant regression of the inflammatory reaction in one case and the disappearance of the tumour in the other one. In conclusion, pharmacomedical therapy can be recommended, if the diagnosis is doubtlessly and a close follow up is established. The risk of anaesthesia and surgical treatment should be weighted against the risk of nonsteroidal antiinflammatory drug treatment. PMID- 12545425 TI - [The familial undescended testis]. AB - BACKGROUND: Testicular maldescent affects 1 % of all 1 year old boys. The observation of familial undescended testis (UDT) was confined to the description of individual cases respectively particular families. The aim of our study was to evaluate the frequency of a positive family history and the frequency of associated urological anomalies in family members and to calculate the risk for new born male individuals to have UDT if a family member is affected. PATIENTS AND METHODS: 74 patients who underwent surgery because of UDT and 374 matched controls without UDT (control group) were interviewed by means of a special questionnaire. We asked for a family history of UDT, as well as for other urogenital anomalies respectively organ alterations (varicocele, hydrocele, hypospadia, testicular cancer, renal anomalies) and reviewed the medical records. For statistical analysis the odds ratio were calculated. RESULTS: 85/374 (22.73 %) of the analysed patients had family members with UDT versus 28/374 (7.5 %) of the control group. Brothers were involved in 37.3 %, fathers in 35.2 %, uncles in 23.5 %, cousins in 16.5 %, great-cousins in 8.2 % and grandfathers in 7.1 %. The familial cluster (risk for UDT in new born male) is 3.6-fold over all (2.306; 5.727), 6.9-fold if a brother and 4.6-fold if the father is affected. The rate of family members with UDT and/or other urogenital anomalies in the patients group was nearly 2-fold higher as in the control group (14 % versus 7.8 %). CONCLUSIONS: In our series the presence of UDT is much more frequently present than described in the literature so far. Due to the high familial cluster and the higher percentage of a positive family history for UDT and/or other urogenital anomalies in patients with UDT, a genetic predisposition is probable. Male descendants show a 3.6-fold increased risk in relation to the normal population, if a family member is already affected. PMID- 12545426 TI - Mutations in the small GTP-ase late endosomal protein RAB7 cause Charcot-Marie Tooth type 2B neuropathy. AB - Charcot-Marie-Tooth type 2B (CMT2B) is clinically characterized by marked distal muscle weakness and wasting and a high frequency of foot ulcers, infections, and amputations of the toes because of recurrent infections. CMT2B maps to chromosome 3q13-q22. We refined the CMT2B locus to a 2.5-cM region and report two missense mutations (Leu129Phe and Val162Met) in the small GTP-ase late endosomal protein RAB7 which causes the CMT2B phenotype in three extended families and in three patients with a positive family history. The alignment of RAB7 orthologs shows that both missense mutations target highly conserved amino acid residues. RAB7 is ubiquitously expressed, and we found expression in sensory and motor neurons. PMID- 12545427 TI - Epimutations in Prader-Willi and Angelman syndromes: a molecular study of 136 patients with an imprinting defect. AB - Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are neurogenetic disorders that are caused by the loss of function of imprinted genes in 15q11-q13. In a small group of patients, the disease is due to aberrant imprinting and gene silencing. Here, we describe the molecular analysis of 51 patients with PWS and 85 patients with AS who have such a defect. Seven patients with PWS (14%) and eight patients with AS (9%) were found to have an imprinting center (IC) deletion. Sequence analysis of 32 patients with PWS and no IC deletion and 66 patients with AS and no IC deletion did not reveal any point mutation in the critical IC elements. The presence of a faint methylated band in 27% of patients with AS and no IC deletion suggests that these patients are mosaic for an imprinting defect that occurred after fertilization. In patients with AS, the imprinting defect occurred on the chromosome that was inherited from either the maternal grandfather or grandmother; however, in all informative patients with PWS and no IC deletion, the imprinting defect occurred on the chromosome inherited from the paternal grandmother. These data suggest that this imprinting defect results from a failure to erase the maternal imprint during spermatogenesis. PMID- 12545428 TI - SCA8 repeat expansion: large CTA/CTG repeat alleles are more common in ataxic patients, including those with SCA6. AB - We analyzed the SCA8 CTA/CTG repeat in a large group of Japanese subjects. The frequency of large alleles (85-399 CTA/CTG repeats) was 1.9% in spinocerebellar ataxia (SCA), 0.4% in Parkinson disease, 0.3% in Alzheimer disease, and 0% in a healthy control group; the frequency was significantly higher in the group with SCA than in the control group. Homozygotes for large alleles were observed only in the group with SCA. In five patients with SCA from two families, a large SCA8 CTA/CTG repeat and a large SCA6 CAG repeat coexisted. Age at onset was correlated with SCA8 repeats rather than SCA6 repeats in these five patients. In one of these families, at least one patient showed only a large SCA8 CTA/CTG repeat allele, with no large SCA6 CAG repeat allele. We speculate that the presence of a large SCA8 CTA/CTG repeat allele influences the function of channels such as alpha(1A)-voltage-dependent calcium channel through changing or aberrant splicing, resulting in the development of cerebellar ataxia, especially in homozygous patients. PMID- 12545429 TI - [Influences of the mobile phase constitution, salt concentration and pH value on retention characters of proteins on the metal chelate column]. AB - The effects of the nature and concentration of salts, pH value and competitive eluent in the mobile phase on the protein retention have been systematically investigated. A mathematical expression describing the protein retention in metal chelate chromatography has been derived. It is proposed that the eluting power of the salt solution can be expressed by the eluent strength exponent epsilon. According to the retention characters of protein under different chromatographic conditions, the interaction between the various metal chelate ligands and proteins is discussed. The protein retention on the metal chelate column is a cooperative interactions of coordination, electrostatic and hydrophobic interaction. For the strong combined metal column with proteins such as IDA-Cu, the coordination is the most important, and the electrostatic interaction is secondary in chromatographic process. However, for the weak combined metal columns with proteins such as IDA-Ni, IDA-Co and IDA-Zn, the electrostatic interaction between the metal chelate ligands and proteins is the chief one, while the coordination is the next in importance. When the mobile phase contains high concentration of salt which can't form complex with the immobilized metal, the hydrophobic interaction between the protein and stationary phase will be increased. As the interaction between the metal chelate ligand and proteins relates to chromatographic operating conditions closely, different elution processes may be selected for different metal chelate columns. The gradient elution is generally performed by the low concentration of salt or different pH for weakly combined columns with proteins, however the competitive elution procedure is commonly utilized for strongly combined column. The experiment showed that NH3 is an excellent competitive eluent. It isn't only give the efficient separation of proteins, but also has the advantages of cheapness, less bleeding of the immobilized metals and ease of controlling NH3 concentration. The interaction between the metal chelate ligand and proteins and the selectivity of metal chelate chromatography can be changed through changing chromatographic conditions. PMID- 12545430 TI - [Chiral separation by capillary electrochromatography on stationary phase adsorbed with protein]. AB - A method based on the adsorption of bovine serum albumin (BSA) on the strong anion-exchange packing (SAX) was applied to prepare chiral stationary phases for capillary electrochromatography. It was found that the electroosmotic mobility only decreased 26.3% when the BSA was adsorbed. The run-to-run reproducibilities of void time and the migration times of D- and L-tryptophan enantiomers were rather good with the relative standard deviation (RSD) values of 0.90%, 0.87% and 0.96%, respectively, when mobile phase containing 7% (volume fraction) acetonitrile. The merit of this system is that the direction of electroosmotic flow is still from cathode to anode, preferable to separate acidic enantiomers. Four anionic enantiomers, tryptophan, ketoprofen, fenoprofen and warfarin, as well as the neutral enantiomers of benzoin were successfully resolved. In order to reduce the analysis time, n-hexanoic acid was added to the mobile phase as a competitive agent. The effect of acetonitrile volume fraction on the resolution of enantiomers was also investigated. PMID- 12545431 TI - [Determination of rutin and quercetin in mulberry leaves by high performance capillary electrophoresis]. AB - Rutin and quercetin are the main effective components of mulberry leaves with the functions of controlling the increase of fat in serum and controlling the formation of arterio-sclerosis. In this article a high performance capillary electrophoretic(HPCE) method was used to separate and determine rutin and quercetin in mulberry leaves collected from different periods, different places and different kinds in Xinjiang. Electrophoretic conditions were as follows: a capillary tube (75 microns i.d. x 57 cm (effective length, 50 cm)), with 10 mmol/L dihydrogen sodium phosphate-20 mmol/L sodium borate containing 15% methanol as the running buffer (pH 8.62) and an applied voltage of 20 kV, at 25 degrees C, detected at a wavelength of 245 nm. Under the optimum conditions, rutin and querctin were separated successfully from other components within 12 minutes. The corrected peak areas of rutin and quercetin increased linearly with the increase of their concentrations in the range of 4.4 mg/L-28.8 mg/L and 1.8 mg/L-145.8 mg/L respectively. The corresponding regression equations for rutin and quercetin were Y = 71.60 rho + 30.51 (r = 0.9991) and Y = 315.17 rho - 929.70 (r = 0.9949) respectively, and the recoveries were 95.64% and 99.36%. The analytical results demonstrate the method is simple, quick and well reproducible, and can be used as a reliable tool for the quality control of mulberry leaves. PMID- 12545432 TI - [Study on the analytical methods of catechins in tea and green tea polyphenol samples by high performance liquid chromatography]. AB - Hypersil BDS C18 and Zorbax SB C18, suitable to separate simultaneously seven kinds of catechins and caffeine, were screened out from seven brands of reversed phase columns. Mobile phase was a solution of methanol-water-acetic acid (or trifluoro acetic acid). Seven kinds of catechins in tea samples from six places in China and three green tea polyphenol(GTP) samples from different producers were separated and determined in 30 min by isocratic and gradient elutions. The effects of mobile phase components and temperature of column on retention parameters of catechins and caffeine are reviewed. Chromatographic conditions and pretreatment methods of samples were optimized. Gallocatechin gallate(GCG) and ( )-catechin gallate(CG) were identified by electrospray ionization mass spectrometry(ESI-MS) and prepared by high performance liquid chromatography for quantitative analysis. The other catechins, (-)-epigallocatechin (EGC), (+) catechin (D-C), (-)-epicatechin(EC), (-)-epigallocatechin gallate(EGCG), (-) epicatechin gallate(ECG) were identified with standards. PMID- 12545433 TI - [Screening of steroidal saponins from the bulbs of Lilium brownii var. colchesteri by combination of high performance liquid chromatography-electrospray ionization mass spectrometry and electron impact mass spectrometry]. AB - With the combination of high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC/ESI-MS) and electron impact mass spectrometry (EI-MS), two steroidal saponins, one compound containing three glycosyls and tigogenin and the other one containing three glycosyls and diosgenin, from the bulbs of Lilium brownii var. calchesteri in China have been screened. In the method, on-line HPLC/ESI-MS allows us to obtain rapidly useful information about the molecular weight and the glycosyl chain of glycoside without the necessity of isolating individual compounds, but little information about steroidal sapogenins. Just with 1 mg to 2 mg of pure sample, off-line EI-MS allows us to acquire useful information about a steroidal sapogenin of saponins, but it is difficult to obtain the molecular ion peak. The combination of HPLC/ESI-MS and EI MS is well suitable for rapidly screening steroidal saponins from plants. PMID- 12545434 TI - [Determination of impurities in a technical chlorpyrifos by gas chromatography with atomic emission detection]. AB - Mass fraction of impurities in a technical chlorpyrifos sample may be obtained by selective determination on carbon (496 nm and 193 nm), chlorine(479 nm), and sulfur (181 nm) channels using gas chromatography with atomic emission detection(GC/AED). According to the formulas from GC/MS and analytical results from GC/AED, mass fraction of compounds may be calculated by internal standard of elements. The results for the same compound obtained from different elemental channels were consistent. This shows the responses of carbon, chlorine and sulfur on AED are independent on molecular structures. Moreover, the reliability of the qualitative results of impurities from GC/MS was proved. If the formulas from GC/MS were wrong, the results from different elemental channels on GC/AED would be different. The relative deviations were in the range of -3.4% to 5.8%. When standards are not available, the results from GC/AED are more reliable than those from GC/FID which usually depend on the area normalization without corrections. Pesticides and other complex organic mixtures may be analyzed reliably and expediently by this method. PMID- 12545435 TI - [Recent advances of chelation ion chromatography]. AB - Chelation ion chromatography (CIC) has the outstanding advantages over the conventional ion chromatography for the analysis of metal ions in complex matrices. In recent years, most research work mainly concentrated on two active areas: 1) to simplify the flow diagram of the CIC system and make it easy to be operated; 2) to develop new high-performance chelation ion chromatographic (HPCIC) methods and new high-performance chelation phases. So, this review presents the principle of CIC method and the advances of the flow diagrams of CIC in the recent decade. The range of applications of high-performance chelation phases used for the separation and determination of metal ions by HPCIC as well as their types and properties are also considered. The high-performance chelation phases include covalently bonded, pre-coated and dynamically coated phases etc. Fifty-three references are cited. PMID- 12545436 TI - [Application of capillary electrophoresis in the analysis of pesticides]. AB - Capillary electrophoresis has proved to be a suitable microseparation technique for the analysis of a wide variety of chiral and achiral pesticides. In this paper, the applications and developments in the field of pesticide separation by HPCE are reviewed with 50 references, including different operating models and chiral agents. Furthermore, the advantage and trend of development are also pointed out. PMID- 12545437 TI - [Study on optimization of the required amount of mobile phase for preparative unit of simultaneous renaturation and purification of proteins]. AB - Based on the stoichiometric displacement model for retention (SDM-R) in high performance liquid chromatography(HPLC), the biopolymer retentions are mainly dominated by displacer's concentration in mobile phase. According to this conclusion, an equation is presented for evaluating the minimum amount of mobile phase used for the preparative purpose with the unit of simultaneous renaturation and purification of proteins (USRPP) by linear gradient elution. Furthermore, the resolution and refolding efficiency of lysozyme were found to be independent of the flow-rate and gradient time when the amount of mobile phase was kept constant. The conclusions were tested by experiments and a good coincidence was obtained. PMID- 12545438 TI - [Application of molecular similarity method in the study of quantitative structure-retention relationship for reversed-phase high performance liquid chromatography of drugs]. AB - Molecular similarity method was applied to the study of quantitative structure retention relationship (QSRR) for reversed-phase high performance liquid chromatography (RP-HPLC) analysis of drugs. Based on a thorough and systematic study on the molecular structures of 162 drugs, molecular similarity method, which transformed molecular structure parameters to similarity variables, combined with artificial neural network for the study of QSRR for RP-HPLC. The good relationship module reflecting molecular structure, solvent strength and capacity factor was established. Molecular similarity method was successfully used to predict capacity factors (k') and the validation results of 7 drugs were satisfactory. The correlation coefficient of test samples was 0.996, and the residual standard error was 0.244(n = 18). The correlation coefficient of validation samples was 0.992, and the residual standard error was 0.131(n = 7). The application of molecular similarity method in the study of QSRR for RP-HPLC of drugs is satisfactory. PMID- 12545439 TI - [Preparation and evaluation of phosphatidylcholine coated on silica as the biomembrane stationary phase]. AB - The method of reverse phase evaporation is used to coat phosphatidylcholine(PC) directly on the surface of silica, which is used as biomembrane chromatographic solid phase to study the interactions between drugs and biomembrane. It was observed that the solid phase coated phosphatidylcholine had a good stability at 20 degrees C-30 degrees C, and the stability would be improved by the presence of appropriate amount of cholesterol in phosphatidylcholine. The content of cholesterol in phosphatidylcholine, the nature of buffer, the concentration of salt in buffer and the pH of mobile phase could all affect chromatographic retention of drugs on the prepared biomembrane column. Six compounds, polyethylene glycol, mannitol, salicylic acid, warfarin, hydrocortisone, and cortisone have been tested. The biomembrane chromatographic stationary phase coated PC with silica as matrices can be simply prepared and it is possible to simulate the human's physiological environment by the biomembrane chromatographic system, so it is a useful method to study drug absorption and distribution in human body. PMID- 12545440 TI - [Simultaneous separation of organic acid and organic salts by electrostatic ion chromatography]. AB - The electrostatic ion chromatographic column was prepared by coating conjugated acid salt micelles on the surface of octadecyl silica stationary phase. Pure water was used as mobile phase, and the conductance detector was connected on line to electrostatic ion chromatograph. The conditions under which organic acid and organic salts were detected were studied. The mechanism for the above separation is discussed. Sodium benzoate and citric acid in Lichee drink were separated and determined. This method is rapid, simple with little interference and good reproducibility without any pollution since the mobile phase is water. This is an environmental friendly analytical method. PMID- 12545441 TI - [Separation of DNA fragments from halobacterium halobium by capillary electrophoresis with non-gel sieving media]. AB - The optimum electrophoresis separation conditions of Lambda DNA/EcoR I + Hind III fragments were investigated using the capillary column DB-1 coated with polysiloxane. When hydroxyethylcellulose (HEC, 1.0%, mass percentage) was used as the only non-gel sieving medium solution, some Lambda DNA fragments could not be separated. After polyvinylpyrrolidone (PVP, 2%, mass percentage) being added into the medium solution, PVP and HEC formed a network colloidal solution in buffer, and changed the network hole size of sieving media. In addition, the results showed that PVP could restrain the adsorption of capillary to DNA, reduced electroosmotic flow, and improved the selectivity of separation. For the first time, all fragments of Lambda DNA marker was separated completely under the same condition of mixed sieving medium solution. The method was applied to separate two groups of DNA fragments of halobacterium halobium, and the base pair number was conjectured. PMID- 12545442 TI - [Direct optical resolution of the enantiomers of biphenyl compounds by high performance liquid chromatography on amylose tris-(3,5-dimethylphenylcarbamate) stationary phase]. AB - A chiral stationary phase(CSP) was prepared by coating amylose tris-(3,5 dimethylphenylcarbamate) onto aminopropylated spherical silica gel. The enantioselective chromatographic separation of a series of biphenyl compounds with antihepatitis activity was performed for the first time on CSP. The effect of alcohol modifiers in mobile phase was investigated. The mobile phases used in the study were hexane modified by ethanol, 1-propanol, 1-butanol or 2-propanol. It was found that the retention times and the separation of the enantiomers of biphenyl compounds on the CSP were dramatically influenced by the nature of alcohols in the mobile phase, and the best alcohol modifier for some compounds was ethanol. The interaction between the stationary phase and samples was also discussed. PMID- 12545443 TI - [Determination of salicin in extract of willow bark by high performance liquid chromatography]. AB - An HPLC method for the determination of salicin in extract of willow bark is described. Chromatographic analysis was carried out on a Kromasil C18, 5 microns column(4.6 mm i.d. x 250 mm) with methanol-0.01 mol/L KH2PO4 buffer (pH 4.01) (15:85, volume ratio) as mobile phase. The detection wavelength was 265 nm. Salicin was extracted from samples with methanol-water(50:50, volume ratio), and centrifuged. Ten microL of supernatant were injected. The average recoveries were from 96.1% to 101.2% (n = 5), and the relative standard deviation (RSD) was 1.43%. The method is simple, rapid and accurate. PMID- 12545444 TI - [LC/MS research on the isomers of special melanoma antigen-encoding gene-2 epitope peptide induced by solvent]. AB - To study the influence of different solvent systems on the isomers of melanoma antigen-encoding gene-2(MAGE-2) epitope peptide synthesized by Marrifield's solid synthesis method, MAGE-2(171-179) epitope peptides were pre-treated using ethanol and methanol systems respectively, and then analysed by RP-HPLC/MS, with dimethyl sulphoxide (DMSO) as control solvent. Results demonstrated that 100% ethanol and 100% methanol solvents could induce the production of isomers of MAGE-2, while 100% DMSO, 50% ethanol and 50% methanol could not. These isomeric peptides could be reversed greatly if treated with trifluoroacetic acid (TFA). The results also showed that the analysis would be distorted and the yield of peptides would decrease dramatically if the peptides were pre-treated with high concentrations of ethanol and methanol before RP-HPLC analysis and preparation. Thus it is suggested that it might be better to decrease the concentration of organic phase in the solvent system for the resolvable hydrophobic peptides, acidified ethanol and methanol might also be better for the highly hydrophobic peptides, and DMSO would be a good solvent for hydrophobic peptides analysis provided that it would not influence the resolving power of chromatography. PMID- 12545445 TI - [Quantitative analysis of sparfloxacin injection by high performance liquid chromatography]. AB - Analytical method of the quantitative determination of sparfloxacin injection by HPLC is described. The analytical conditions were as follows. A Waters Symmetry C18(5 microns, 150 mm x 3.9 mm i.d.) column was used as the analytical column. The detection wavelength was UV-298.8 nm. The column temperature was 30 degrees C. The mobile phase was 0.2% KH2PO4 buffer (pH 3.2)-CH3CN-CH3OH (80:15:5, volume ratio) and the flow-rate was 1.0 mL/min. The injection volume was 10 microL. The linear range (the peak area vs. the mass concentration of sparfloxacin) was from 39.94 mg/L to 199.68 mg/L, and the correlation coefficient was 0.9999. The average recovery of sparfloxacin was 100.1% (n = 5), and its RSD was 0.72%. The RSDs of continuous injections, within day injections per 2 hours and between day injections in three days were 0.19%, 0.14% and 0.13% respectively. The above analytical results show that this method has good precision and stability. It is a rapid, sensitive and accurate method for the analysis of sparfloxacin. PMID- 12545447 TI - [Separation of two isomers in the products of condensation reaction with trimethoxybenzaldehyde as raw material and quick determination of the concentration of its intermediates by high performance liquid chromatography]. AB - The content ratio of the two isomers in the products of condensation reaction with trimethoxybenzaldehyde as raw material is the key factor to improve the productivity of trimethoprim(TMP). A quantitative method using Nova-Pak C18 column (3.9 mm i.d. x 150 mm, 4 microns) and tetrahydrofuran(THF)-water (30:70, V/V) as mobile phase with flow rate of 1.0 mL/min and UV 320 nm detector was established. A good linearity was obtained in the concentration range of 0.01 mg.L-1-10 mg.L-1. The detectable limits of trimethoxybenzaldehyde and alpha (3,4,5-trimethoxy-benzal)-beta-methoxy-propionitrile were 2.0 micrograms/L and 1.0 microgram/L respectively. The method has been used to analyze real samples with satisfactory results. PMID- 12545446 TI - [Determination of puerarin in three traditional Chinese patent medicines by high performance liquid chromatography]. AB - A simple and accurate reversed-phase high performance liquid chromatographic method to determine puerarin in three traditional Chinese patent medicines, namely "Xiao'er qingganling" tablet, "Buchang xinnaoxintong" capsulae and "Ganmao qingre" granule is described. "Xiao'er qingganling" tablet and "Buchang xinnaoxintong" capsulae were extracted with methanol in an ultrasonic bath and "Ganmao qingre" granule was extracted with water and ethanol. After being centrifuged and filtered, the samples were separated on an ODS column with a mobile phase of 100 mmol/L ammonium acetate buffer (pH 5.0)-methanol(75:25, volume ratio). The flow rate was 0.8 mL/min, and the detection wavelength was 250 nm. The calibration curve was linear in the range of 2 mg/L-20 mg/L puerarin (r = 0.9999). The average recoveries were 99.0%, 93.4% and 97.5% for "Xiao'er qingganling" tablet, "Buchang xinnaoxintong" capsulae, "Ganmao qingre" granule respectively. The average contents of puerarin in the medicines were 3.48 mg/g, 1.08 mg/g and 1.52 mg/g (sucrose), and the relative standard deviations were 1.9%, 1.4% and 1.4% respectively. PMID- 12545448 TI - [Addition internal standard method in chromatographic quantitative analysis]. AB - Internal standard method is a conventional chromatographic quantitative method which requires one or several internal standards added. The internal standard component must not be contained in the sample and need a good separation between the internal standard and sample components. In many cases selecting an internal standard is not convenient or even restricted by the seperation of components. In this paper, we try to combine the internal standard method and the addition method to form a new chromatographic quantitation method named addition internal standard method. The principles of addition internal standard method are suitable to not only chromatographic quantitation but also polarography etc. The related theory and foundation of the method are defined. The operation steps and the conditions suitable to the method are discussed. The advantages and disadvantages of this method are explained in detail. PMID- 12545449 TI - [Extraction of residual formaldehyde in polymer complex and high performance liquid chromatographic analysis]. AB - This paper presents a simple and accurate method for analyzing formaldehyde in polymer complex by headspace extraction derivatization and HPLC analysis. The sample preparatio of formaldehyde in polymer complex was based on a simple thermodynamic equilibrium in a closed and thermostat jar, and the gaseous formaldehyde in equilibrium could be absorbed and extracted by the liquid in the bottom of the jar. The formaldehyde was derivatized with 2,4-dinitrophenyl hydrazine before chromatography. The influences of temperature, equilibrium time, sample quantity and geometric form on the extraction efficiency were studied. The extraction and HPLC conditions were optimized. The limit of detection (LOD) was 0.1 mg/kg, and the relative standard deviation (RSD) was below 5%, and the recoveries were between 96%-103%. As a result, this method would meet the demands for analyzing microamounts of residual formaldehyde in polymer complex. PMID- 12545450 TI - [Determination of solanesol in the extracts of tobacco leaves by high performance liquid chromatography]. AB - A method for the determination of solanesol in the extracts of tobacco leaves by high performance liquid chromatography has been developed. The column used was Shim-pack CLC-SIL(5 microns, 5 mm i.d. x 150 mm) and the UV detector was set at 215 nm. The eluent was a mixture of n-hexane and isopropyl alcohol (98:2, V/V). As the results showed, the linear range was 1 microgram-10 micrograms(r = 0.9997), the average recovery of solanesol was 98.1% and the relative standard deviation was 1.9%. This method is fast, accurate and reproducible. PMID- 12545451 TI - [Separation and determination of scandium, tin and aluminum by reversed-phase ion pair high performance liquid chromatography]. AB - The reversed-phase ion pair high performance liquid chromatographic determination of scandium, tin and aluminum with diammonium stilbene-4,4'-bis(1-azo)-3,4 dihydroxybenzene-2,2'-disulfate as precolumn chelating reagent on Spherex C18 column has been investigated with the detection wavelength of 500 nm. Various parameters such as pH, reagent concentration, ion pair reagents, interference of foreign ions have been studied. The mobile phase was methanol-water (40:60, V/V) containing 20 mmol/L acetate buffer of pH 6.0 and 10 mmol/L sodium dodecyl sulfonate. Three complexes could be completely separated within 17 min. The method has been successfully used for the determination of the three metals in mineral samples, with recoveries of 98%, 101%, 103% respectively, and RSD was less than 3.6%. Beer's Law was obeyed over the concentration range 0.01 mg/L-3.80 mg/L, 0.06 mg/L-4.50 mg/L and 0.04 mg/L-3.40 mg/L respectively. The detection limits were 0.9 microgram/L, 1.0 microgram/L and 1.2 micrograms/L for scandium, tin and aluminum when the ratio of signal to noise was 3. Most of other metals did not interfere with the determination. The present method is sensitive and selective. PMID- 12545452 TI - [Analysis of fatty acids in rat liver peroxisomes]. AB - Peroxisomes of rat liver from normal and di (2-ethylhexyl) phthalate (DEHP) treated animals were isolated by density gradient centrifuge method. The fatty acids in the liver peroxisomes were extracted with chloroform and methanol, and were esterified with boron trifluoride and methanol. The fatty acids methyl esters were separated on SPB-1 (similar to SE-30, OV-101) capillary column, and were determined using heptadecanoic acid (C17:0) as an internal standard for the eleven fatty acids (C10:0-C22:6) in these peroxisomes by gas chromatography. The ratio of unsaturated fatty acids to total fatty acids in the control was significantly less than that in DEHP-treated animals (P < 0.05), but there was no difference between the control and DEHP-treated animals for the content of total fatty acids and the ratio of long chain fatty acids to total fatty acids. The results show that DEHP-treatment can alter the fatty acid composition of peroxisome. The membrane structure of the treated peroxisome was different to that of the control. PMID- 12545453 TI - [Determination of cholesterol and alpha-tocopherol in eggs by capillary gas chromatography]. AB - A simple and rapid method has been developed for simultaneous determination of cholesterol and alpha-tocopherol in eggs. The method involves alcoholic KOH saponification of the samples, extraction with petroleum ether, and determination by wide-bore capillary gas chromatography. The recoveries of cholesterol and alpha-VE in eggs were 91.0%-93.3% and 82.5%-95.0%, with RSDs of 2.7%-3.1% and 4.9%-6.9% respectively. PMID- 12545454 TI - [Effects of composition of mobile phases on retention behavior of solutes in hydrophobic interaction chromatography]. AB - The retention behavior of aromatic alcohol homologues in hydrophobic interaction chromatography (HIC) was investigated for the first time. The differences in retention behavior between proteins and small solutes, and the eluted range and ability of different salts to small solutes and biopolymers, are elucidated with the parameter Z of the stoichiometric displacement retention model. The retention of aromatic alcohols follows the homologue rule. By the comparison of the changes of Z in different salt solutions, it is found that for small molecules the salt type only changes the hydration degree; but for proteins, it not only changes the hydration of proteins and stationary phase, but also affects the conformation of protein molecules near the contact regions between the protein and the stationary phase surface. PMID- 12545455 TI - [Preparation and evaluation of alkyl ether-bonded phase for reversed-phase high performance liquid chromatography]. AB - A new silane coupling agent, beta-(3,4-epoxycyclohexyl)ethyltrimethoxy silane, was for the firsty time used to react with octanol, then the intermediate product was coupled onto porous silica to obtain a novel bonded phase for reversed-phase high performance liquid chromatography (RP-HPLC). Characterization of prepared packing was carried out with elemental analysis, solid-state 13C nuclear magnetic resonance spectrometry (NMR), Fourier transform infrared(FT-IR) spectroscopy. Chromatographic evaluations were carried out by using a mixture of organic compounds and methanol-water as the binary mobile phase. The results showed that the stationary phase has excellent chromatographic properties and resist to hydrolysis at a pH value between 2.5 and 7.5. The silanophilic activity of the OEBP was weakened because of the existence of the cyclohexyl group in the packing. It can be efficiently used for the separation of basic compounds. PMID- 12545456 TI - [Determination of coenzyme Q and alpha-tocopherol in mouse plasma and tissue mitochondria by liquid chromatography with electrochemical detection]. AB - A convenient and reliable liquid chromatographic method with electrochemical detection was developed for the simultaneous determination of reduced coenzyme Q(ubiquinol), oxidized coenzyme Q(ubiquinone) and alpha-tocopherol in various tissues. The compounds of interest were separated on a reverse-phase C18 column under the mobile phase of 7 g/L NaClO4 in ethanol-methanol-70% HClO4 (800:200:1, volume ratio) and detected simultaneously by using a series of detection cells. Electrochemical guard cell (upstream of the injector) was used for eliminating interference for mobile phase. Ubiquinones were reduced in the -550 mV conditioning cell (downstream of the column). The resultant ubiquinols and original ubiquinols and alpha-tocopherol existing in the sample were easily oxidized in the 150 mV analytical cell. The linear range of the compounds were from 5 micrograms/L to 200 micrograms/L with correlation coefficients from 0.9976 to 0.9999. The detection limits were from 0.23 microgram/L to 0.68 microgram/L. The method was applied satisfactorily to the simultaneous determination of the amounts of ubiquinol-9, ubiquinol-10, ubiquinone-9, ubiquinone-10 and alpha tocopherol in mice plasma and tissue mitochondria. PMID- 12545457 TI - [Determination of anions in liquid crystal samples by UV-photolysis-ion chromatography]. AB - A new method for the determination of anions in liquid crystal samples by UV photolysis-ion chromatography was developed. The sample pretreatment method, the photolytic conditions and the chromatographic parameters were optimized. The results indicate that there have five anions (F-, Cl-, NO2-, Br-, I-) in the liquid crystal sample. By the filtration with the Dionex OnGuard RP and P cartridges, most of the organic compounds in the photolyzed sample solution can be removed. In the process of photolysis, the efficiency can be remarkably enhanced by the adding of H2O2. Iodide in the liquid crystal sample can be accurately determined by adding of 0.1 mmol/L NaOH. In the five anions, fluoride, choloride, nitrite and bromide can get their maximal responses when the photolysis time is 2 h. With the strongest reducibility, iodide only needs 1.5 h to get the highest response. The separation of the five anions was accomplished with Dionex IonPac AS16 column and the determination can be completed in 25 min. The recoveries for the five anions were between 85.1% and 111.5%. PMID- 12545458 TI - [Advances in solid phase microextraction coupled with high performance liquid chromatography]. AB - Solid phase microextraction (SPME) is a solvent-free technique with high extraction efficiency and easy to perform automatically. It can be coupled with high performance liquid chromatography to perform efficient analysis of compounds with high polarity. The advances in solid phase microextraction coupled with high performance liquid chromatography, including its theory, interfaces, coating materials and applications, are reviewed with 36 references. PMID- 12545459 TI - [Laser-induced fluorescence detection in micro-column separation]. AB - Laser-induced fluorescence detection (LIFD) has been used extensively in micro column separation due to its high sensitivity and selectivity. A review is presented on the present status and the trends of development of LIF detector with 52 references cited. The detection cells of the LIFD are mainly discussed. PMID- 12545460 TI - Derivatization in liquid chromatography/mass spectrometric analysis of neurosteroids. AB - Liquid chromatography/mass spectrometry (LC/MS) is now considered to be the most promising analytical method for the determination of biological substances, especially nonvolatile or highly polar substances. However, some compounds do not show enough sensitivity in LC/MS and soft-ionization methods commonly used in LC/MS, such as electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI), sometimes do not give satisfactory structural information. This report presents an overview of the derivatization methods in the LC/MS analysis of neurosteroids or neuroactive neurosteroids, which are synthesized and accumulated in the nervous system. The derivatization of pregnenolone 3-sulfate, one of these steroids, with 4-(N,N-dimethylaminosulfonyl)-7-hydrazino-2,1,3 benzoxadiazole gave a satisfactory sensitivity during the quantitative analysis using LC/ESI-MS. The obtained results are much lower than those previously obtained using gas chromatography/mass spectrometry or radioimmunoassay. On the other hand, the derivatization to acetate was useful for the treatment of labile catechol estrogens in rat brains and gave enough structural information in LC/APCI-MS, which confirmed the existence of catechol estrogens in mammalian brains. PMID- 12545461 TI - [Study on the rule of retention value in weak acid anion chromatography]. AB - In chromatography, if analytes are strong acid anions or strong alkali cations, the dependence of a logarithm of the adjusted retention value on the logarithm of the concentration of the eluent is given by log tR' = const - (j/i).log C, where j is the charge of the analyte ion, i is the charge of the eluent ion, const is a constant that depends on the column, eluent and analyte ion. But in weak acid anion chromatography, existing forms of the sample anion change with the change of the concentration of the eluent. So the study on the relationship between retention value of weak acid anion and concentration of the eluent is more difficult. Relationship between adjusted retention value of some weak acid anion and concentration of the eluent was studied with sodium hydroxide as eluent. The nonlinear fitting function has been set up. We transform it into multiple linear fitting to give the answer. The correlation coefficients for phosphate defined by the function were over 0.99. The relative errors with the use of function were below 5%. PMID- 12545462 TI - [Synthesis and evaluation of ethylstyrene-divinylbenzene-coated titania for high performance liquid chromatography]. AB - Ethylstyrene-divinylbenzene-coated TiO2 (ES-DVB-TiO2) was synthesized by at first bonding of vinyltriethoxysilane with hydroxyl group on titania and secondly by addition polymerization of vinyltriethoxysilane and ethylstyrene-divinylbenzene. The hydrophobic selectivity, absolute mobile phase sensitivity, and stability of the stationary phase were evaluated. Finally, several kinds of practical separations were tested on ES-DVB-TiO2. These examples showed that ES-DVB-TiO2 synthesized in our laboratory was a good reversed-phase stationary phase. PMID- 12545463 TI - [Diol column as stationary phase for high performance liquid chromatographic analysis of carbohydrates in drinks with evaporative light scattering detection]. AB - A high performance liquid chromatographic method with a diol column and evaporative light scattering detector (ELSD) was established for the direct analysis of fructose, glucose, sucrose, maltose and raffinose in mixture. A separation column (Lichrospher 100 Diol, 250 mm x 4.0 mm i.d., 5 microns, Hewlett Packard, USA) and a guard column (Zorbax Rx-SIL, 12.5 mm x 4.6 mm i.d., 5 microns) were used. The mobile phase was a mixture of dichloromethane-methanol (3.2:1, volume ratio). Regression equations revealed linear relationship (correlation coefficients: 0.995-0.999) between the mass of carbohydrates injected and the peak area of carbohydrates detected by ELSD. The detection limits of ELSD (S/N = 3) were about 0.20 microgram for all carbohydrates. This system could be used for the routine analysis of simple carbohydrates in some common drinks on market. PMID- 12545464 TI - [Determination of thyroxine enantiomers in human plasma with normal high performance liquid chromatography-chiral complex exchange mobile phase]. AB - An HPLC method was developed with normal HPLC-chiral complex mobile phase for the separation and determination of D- and L-thyroxine enantiomers (D- and L-T4) in human plasma. The method includes extraction of thyroxine from plasma and separation of thyroxine enantiomers on HPLC silica column with chiral eluent containing L-proline, cupric acetate and triethylamine (TEA). The sensitivity of the method was 0.1 mg/L. The precisions of inter-day and intra-day, linearity, extraction recovery, and stability of T4 enantiomers in plasma and in deproteinized plasma were determined for the validation of the method. Baseline enantioseparation of the compounds containing D- and L-T4 was achieved. Meantime we determined the concentrations of D-T4 and L-T4 in plasma of 15 volunteers with euthyroid, hypothyroid and hyperthyroid symptoms. Liquid chromatographic method based on the described procedure was useful for the determination of D- and L thyroxine in patient plasma and for pharmacokinetics investigation. PMID- 12545465 TI - [Determination of optical purity of SR-bioallethrin enantiomers by chiral high performance liquid chromatography]. AB - A chiral stationary phase(CSP) was prepared by coating cellulose-tris (3,5 dimethylphenylcarbamate) onto aminopropylated spherical silica gel. On the CSP, the chiral separation of the bioallethrin enantiomers has been investigated and under the optimum conditions the optical purity of three samples of SR bioallethrin enantiomers was determined by peak area. The results show that the method established is very ideal for determining the optical purity and evaluating the quality of the samples. PMID- 12545466 TI - [Analysis of surface-active substances in Sapindus mukurossi by high performance liquid chromatography-mass spectrometry]. AB - A high performance liquid chromatography-atmospheric pressure ionization mass spectrometry method has been developed for the analysis of surface-active substances (hederagenin saponins and sesquiterpene oligoglycosides) in the extracts of the pericarp of Sapindus mukurossi. The method consists of the separation of surface-active substances using C18 HPLC column, followed by detection using a diode-array detector at 210 nm and then on-line mass spectrometry. Hederagenin saponins and sesquiterpene oligoglycosides were characterized as [M - H]- or [M + Na]+. Based on the relative molecular mass, established by mass spectrometry and the structure induced by in-source CID technology, three components that had not been reported in Sapindus mukurossi before were identified. Several surface-active substances were obtained by means of semi-preparative HPLC. Their structures were further confirmed by NMR spectrometry as mukurozi-saponin Y2, mukurozi-saponin X, mukurozioside I a and mukurozioside II a. PMID- 12545467 TI - [Determination of 4-aminobutyric acid in pumpkin powder by high performance liquid chromatography]. AB - A method for the separation and determination of the 4-aminobutyric acid in pumpkin powder by high performance liquid chromatography (HPLC) with post-column derivatization and fluorescence detection is described. The operating conditions were cation exchange resin column (30 cm x 0.4 cm i.d.) with gradient elution of buffer solutions A [19.6 g sodium citrate dihydrate and 1.0 g phenol dissolved in 1 L water, pH(3.15 +/- 0.02)] and B [21.0 g sodium nitrate and 1.5 g boric acid dissolved in 1 L water, pH(9.70 +/- 0.02)] as mobile phase at a flow rate of 0.5 mL/min, and a column temperature of 62 degrees C, with detection wavelength lambda ex = 338 nm, lambda em = 425 nm. The retention time of 4-aminobutyric acid was 34.77 minutes. The average recovery was 99% and the coefficient of variation was 1.35%. This method is simple, rapid and sensitive. PMID- 12545468 TI - Simultaneous determination of sixteen phenylurea herbicides in water by high performance liquid chromatography and solid phase extraction. AB - A high performance liquid chromatography (HPLC) procedure for simultaneous determination of sixteen phenylurea herbicides in water is described. A Lichrospher 100 RP-18e column and spectrophotometric detection at 240 nm were used. Adequate retention was achieved with a mobile phase of acetonitrile/water solution with gradient elution. The HPLC analysis time was less than 20 min. The herbicides were isolated from water samples by using a single solid phase extraction procedure with C18 solid-phase columns. An enrichment factor of 1,000 was achieved. Recoveries were in the range between 87.8% and 103.7%. The detection limits of the whole procedure achieved were less than one-tenth of the maximum limit permitted by the European Community standard for drinking water. PMID- 12545469 TI - [Determination of acyclovir in mouse plasma and tissues by reversed-phase high performance liquid chromatography]. AB - The aim of this study was to establish an high performance liquid chromatographic method for determining acyclovir (ACV) concentration in mouse plasma and tissues. A solution of 0.25 mL 60 g/L perchloric acid and 0.25 mL acetonitrile was added into 0.2 mL plasma or 0.2 g tissues to precipitate proteins. Following centrifugation, the supernatant obtained was injected into a reversed-phase column. Operating conditions were Hypersil ODS column(250 mm x 4.6 mm i.d., 5 microns), methanol-water-acetic acid(1:99:0.5, volume ratio) solution as mobile phase at a flow rate of 1.5 mL/min, UV detection at 252 nm. The detection limit of ACV concentration in plasma was 20 micrograms/L and that in tissues was 50 ng/g. The standard curves for ACV were linear in plasma and homogenate of tissues (r > 0.99). The precision of the method was good and the recoveries of ACV were higher than 97.5%. So this method is rapid, accurate and convenient for determination of ACV concentrations in plasma and tissues. PMID- 12545470 TI - [Preparation and evaluation of in-situ synthesis zeolite gas-liquid modified columns]. AB - A new kind of NaA zeolite column has been prepared by in-situ synthesis technique. In comparing with traditional coating method, the in-situ NaA zeolite carrier is much more uniform and rigid. Several kinds of zeolite gas-liquid modified columns have been prepared with this in-situ technology, which showed satisfactory features in separation property and thermal stability. The successful utilization of mini-length and wide-bore columns showed good practical prospect of this new kind of zeolite column. PMID- 12545471 TI - [Determination of volatile organic compounds in atmospheric environment]. AB - It is well known that volatile organic compounds (VOCs) are the main photochemical pollutants and ozone precursors of the photochemical smog. Investigation of photochemical pollution in the ambient air must focus on VOCs, but the concentration of VOCs in ambient air is in a very low level (10(-9)-10( 12), volume fraction), so there are difficulties in the determination of VOCs. In this work, based on the TO14A and TO15 methods recommended by the Environmental Protection Agency of United States, an improved method for the determination of fifty-six VOCs, mainly O3 precursors, in atmospheric environment was developed. Operating conditions of VOCs preconcentrator, gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) were optimized. Air sample was first frozen by liquid nitrogen, and then H2O and CO2 were eliminated in the VOCs preconcentrator. The preconcentrated VOCs sample was injected to GC and detected by MS or hydrogen flame ionization detector (FID). The C2-C10 hydrocarbons were separated effectively in capillary columns under the high concentration of CO2. The detection limits were 0.1 microgram.m-3 and the relative standard deviations were in the range from 2.57% to 9.82%. This method has been used for the determination of VOCs in real samples. The results were satisfactory. PMID- 12545472 TI - [Separation of xylose and xylitol in fermentation liquid by capillary zone electrophoresis]. AB - Xylitol may be produced with microbial fermentation technology when xylose is used as the raw material. It is important in scientific research and production fields concerned with xylose and xylitol to develop the method of separation and detection. The anionic complexes with strong UV absorption at 195 nm will form by dissolving xylose and xylitol in borax solution. They may be separated with borax buffer by use of capillary zone electrophoresis. It was shown that the resolution between xylose and xylitol gradually increased with the increase of borax concentration, but its maximum concentration was 130 mmol/L at room temperature. The resolution depended on the pH of running buffer with a maximum at pH 9.55. The resolution was independent of cetyltrimethylammonium bromide when its concentration was between 4 x 10(-6) mmol/L-8 x 10(-4) mmol/L. So the optimum conditions were as follows: borax concentration of 130 mmol/L, pH 9.55 and cetyltrimethylammonium bromide concentration of 5 x 10(-5) mmol/L in running buffer; separating voltage of -12 kV; column temperature of 25 degrees C. Xylose and xylitol could be separated on base line in 6 min under these conditions. Samples from fermentation process and the recoveries of spiked samples were determined. The relative standard deviations of the results were between 1.42% 3.11% for xylose, and 0.62%-1.32% for xylitol. The recoveries were between 96.0% 108.0% for xylose and 94.0%-109.0% for xylitol. PMID- 12545473 TI - [Separation and determination of pseudoephedrine in bufferin cold tablet by capillary electrophoresis with hydropropyl-beta-cyclodextrin as chiral selective reagent]. AB - A method for the separation and analysis of the chiral pseudoephedrine enantiomers using capillary electrophoresis was established. The buffer solution for separation was 25 mmol/L Tris-phosphate, including 38 mmol/L hydropropyl-beta cyclodextrin with pH value of 2.65. (1S, 2S)(+) Pseudoephedrine in Bufferin Cold tablet was determined. The method has good precision, recovery and linear relationship. PMID- 12545474 TI - [Separation and purification of an endogenous inhibitor of sodium pump from chansu by thin-layer chromatography and reversed-phase high performance liquid chromatography]. AB - An endogenous inhibitor of the sodium pump from the Chinese medication Chansu was purified. The dry substance Chansu was extracted with methanol. The dry residue dissolved in water and filtered subsequently through membrane filters with the exclusion size of 1000 Da, 3000 Da and 10000 Da in a Filtron Pro Vario-3-System and applied to thin-layer chromatographic plate made of Silica gel 60 F254 + 366 developed with a mixture of CHCl3-MeOH-H2O(75:20:5, volume ratio). The fractions with Rf 0.55 inhibiting the sodium pump were purified on an HPLC C18-RP column using a linear H2O-methanol gradient with 220 nm and 300 nm DAD detection. The bioactivity was measured by 86Rb-uptake into human red blood cells. The results showed that a low molecular weight, water soluble compound, which inhibited the sodium pump activity in the red blood cells and had a maximum absorbance at 250 nm was isolated from the Chinese medication Chansu. Several mg of the compound in pure state could be obtained from 1 kg Chansu. It was different from ouabain and proscillaridin A in chemical structure, because ouabain and proscillaridin A show a UV maximum absorption at 220 nm and 300 nm, while the new inhibitor at 250 nm. PMID- 12545475 TI - [Determination of phenazine-1-carboxylic acid in anti-fungal agent M18 by high performance liquid chromatography]. AB - A reversed-phase HPLC method for the determination of phenazine-1-carboxylic acid (PCA) in antifungal agent M18 is established. The mobile phase was a mixture of MeOH-5 mmol/L phosphate buffer (pH 5.0) (60:40, volume ratio). The flow rate was 1.0 mL/min, and the detection wavelength was 248 nm. The linear range and detectable limit were 50 mg/L-500 mg/L and 30 mg/L respectively. The recovery was 97.53% and RSD was 1.5%. The method of PCA extraction and detection has proven to be much faster, simpler, more sensitive, accurate and reproducible than those reported already. The assay results can be used as a very important criterion for large-scale production. PMID- 12545476 TI - [Improvement of a method for separation and determination of amino acids]. AB - A method for separation and determination of amino acids using pre-column derivatization with 6-aminoquinoline-N-hydroxysuccinimido-carbamate (AccQ. Tag) was improved. The technique developed for rapid separation and determination of amino acids by AccQ. Tag method is described. The 18 amino acids and taurine are rapidly separated and determined by AccQ. Tag special chromatographic column(3.9 mm i.d. x 150 mm) within 17.5 min(the analysis period is 22.5 min) by changing the flow rate from 1.0 mL/min to 2.0 mL/min. In place of the AccQ. Tag special chromatographic column, the Nova-Pak C18 column(3.9 mm i.d. x 150 mm), Nova-Pak C18 column(4.6 mm i.d. x 150 mm), Symmetry C18 column(3.9 mm i.d. x 150 mm) or Waters Xterra RP-18 column, can be used to separate and determine the amino acids rapidly in the same manner. But among them the Symmetry C18 column is the best substitute for AccQ. Tag column. The separation time is 14.5 min and the analysis period is 20.0 min only. The price is much cheaper than the AccQ. Tag column. PMID- 12545477 TI - [Determination of polyphenols in tobacco by solid phase extraction and high performance liquid chromatography]. AB - A high performance liquid chromatographic (HPLC) method for the determination of polyphenols in tobacco was studied. Polyphenols can be extracted from tobacco sample by refluxing with 80% (volume fraction) methanol, then subjected to degrease by solid phase extraction with Sep-Pak-C18 column. Chlorogenic acid, rutin, scopoletin, caffeic acid, scopolin and other polyphenols can be separated on a Nova-Pak-C18 column (3.9 mm i.d. x 150 mm) gradiently eluted with methanol and 0.05 mol.L-1 potassium dihydrogen phosphate buffer solution as mobile phase at a flow-rate of 0.5 mL.min-1. Polyphenols were determined at the maximum wavelength of each polyphenol, for chlorogenic acid 326.1 nm, rutin 354.8 nm, scopoletin 344.0 nm, caffeic acid 323.7 nm and scopolin 365.2 nm by photodiode array detector. The main polyphenols in tobacco can be identified with UV spectroscopy. The recoveries of tobacco polyphenols were 94%-105%, and the relative standard deviations were 1.28%-1.49%. This method can be applied to the determination of polyphenols in tobacco with satisfactory results. PMID- 12545478 TI - [Study on chemical constituents of the essential oil from Zanthoxylum Bungeanum Maxim by gas chromatography-mass spectrometry]. AB - The essential oil in Zanthoxylum Bungeanum Maxim was extracted by steam distillation and the distilled liquid was collected by passing through chloroform. The aqueous phase was extracted with chloroform sufficiently. The chloroform solution containing the constituents of essential oils was analyzed by gas chromatography-mass spectrometry(GC-MS). Seventeen peaks were separated by gas chromatography, and 16 of them were identified by MS with "Nist98.L" Mass Spectral Data Registry. The structures of the constituents were further verified by manual analysis. The identified constituents accounted for 97% of the peak areas of the essential oils on total ion chromatogram. The major chemical constituents of them are C9, C10 alcohols and alkenes. PMID- 12545479 TI - [Determination of carfentrazone-ethyl(F8426) residue in soil and wheat by wide bore capillary gas chromatography]. AB - An efficient method for determining F8426 of Affinity 40 DF in soil and wheat is described. F8426 was extracted from the soil and the wheat sample with acetone water (80:20, volume ratio), and the extract was partitioned with petroleum ether. Then the upper layer was concentrated and cleaned up on a micro-column of Florisil and activated carbon mixture. Finally F8426 was separated from other interfering components through wide-bore capillary gas chromatographic column (OV 1701, 12 m x 0.53 mm i.d.), and was detected by electron capture detector (ECD). The lowest detection limit for F8426 was 0.02 ng. The lowest detectable concentrations in wheat and soil was 2 micrograms/kg and 1 microgram/kg respectively. The average recovery and coefficient of variation of this method were 89.60%-97.53% and 4.42%-8.67% respectively. This method is simple, sensitive and suitable for residue analysis of F8426. PMID- 12545480 TI - [Application of wide-bore capillary gas chromatography in the synthesis of special esters of methacrylic acid]. AB - A simple, rapid and accurate method for separating and analysing components in the synthesis of special esters of methacrylic acid by gas chromatography is reported. An SE-54 wide-bore capillary column of 50 m x 0.53 mm i.d. x 0.6 microns was used. The esters include dimethylaminoethyl methacrylate(DMAM), n butyl methacrylate(BMA) and ethylene glycol dimethacrylate(EGDMA). Crude materials such as methyl methacrylate(MMA) and N, N-dimethyl ethanol amine (DMEA) were also analysed. The recoveries were 99.71%-102.89%, and the RSDs of the main components were below 0.1%. The method can be used to control the production of the esters. PMID- 12545481 TI - [Determination of 1,1'-peroxydicyclohexylamine by gas chromatography]. AB - A method for the determination of 1,1'-peroxydicyclohexylamine and cyclohexanone by gas chromatography was investigated. The conditions were optimized by changing the stationary liquid, internal standard and injector temperature. The gas chromatographic conditions were as follows: flame ionization detector; 10% OV-101 column; the injector temperature, 140 degrees C; temperature programming; internal standard, dodecane. All components and internal standard were separated in 11 min. The method is simple, rapid, sensitive and reproducible. PMID- 12545482 TI - [Preparation and evaluation of wide-bore stainless-steel packed capillary liquid chromatographic columns]. AB - A zero-dead volume column end and a conical shaped slurry reservoir for wide-bore stainless-steel packed capillary liquid chromatographic columns were designed and evaluated. A detailed procedure for the preparation of reversed-phase stainless steel packed capillary columns with 0.5 mm-1.0 mm i.d. is described. The influences of the column length and the packing material on the column performance have been studied. Columns were evaluated by the reduced plate height vs reduced velocity curve and the peak asymmetric factor. Experimental results showed that the column efficiency and the reproducibility were better than 75% of theoretical value and 6% RSD, respectively. Separations of antiepileptic drugs and chlorinated benzenes are demonstrated. PMID- 12545483 TI - [Preparation and evaluation of amide-octyl-bonded phase for the separation of basic substances in reversed-phase liquid chromatography]. AB - A new bonded-phase, amide-octyl-bonded phase (AOBP), with an internal polar functional group-amide was firstly prepared in domestic by two-step reaction process. The amino phase was formed in the first step and further modified in the second step by attaching an alkyl chain via reaction of an acid chloride with the amino ligand. Characterization of prepared packing was carried out with elemental analysis and Fourier transform infrared (FTIR) spectrometry. Hydrophobicity, selectivity and silanophilic activity of AOBP were evaluated by using organic components including acidic, bascic and neutral analytes and methanol-water as binary mobile phase. The applicable ranges of pH and stability of AOBP were also evaluated. The results showed that AOBP has excellent chromatographic properties and resistance to hydrolysis between pH = 2.5-7.5, It can be used for the separation of basic solutes efficiently. PMID- 12545484 TI - [Preparation and chromatographic characteristics of octadecyl bonded titania stationary phase for high performance liquid chromatography]. AB - Porous titania microspheres (surface area: 36.7 m2/g, average pore volume: 0.30 mL/g, average pore diameter: 32.2 nm, average diameter: 3.5 microns) with a very narrow particle size distribution, were synthesized by refluxing polymerized induced colloid aggregation (PICA), with octadecyltrimethoxysilane in toluene as a solvent for 8 h, and octadecyltitania stationary phase (ODT) was obtained. The carbon content and hydrophobic selectivity of the stationary phase were 2.87% (i.e. 0.66 mumol/m2) and 0.4638, respectively. ODT showed good chromatographic performances when it was used to separate the neutral and basic solutes using methanol solution as a mobile phase. PMID- 12545485 TI - [Simultaneous purification and renaturation of recombinant human interferon-alpha expreessed by E. coli by high-performance hydrophobic interaction chromatography]. AB - One-step simultaneous purification and renaturation of interferon-alpha from the inclusion body of E. coli expreessed by high-performance hydrophobic interaction chromatography(HPHIC) are presented. The inclusion body was dissolved by 8 mol/L guanidine hydrochloride. The sample lysis was centrifuged and the solution was renaturated by three different methods, i.e. hydrophobic interaction chromatography, dilution and dialysis. The total activity yield renaturation by hydrophobic interaction chromatography was 2 times and 2.6 times as much as that by dilution and by dialysis. The purity of the target product was 95% according to high-performance size exclusion chromatography and the specific activity was 1.8 x 10(8) IU/mg. The method developed is simpler and more efficient than others. PMID- 12545486 TI - [Contrastive study of two methods("programmed temperature vaporization with back flushing" and "head space") for light hydrocarbon analysis]. AB - Light hydrocarbon analytical method of "PTV with Back Flushing" presented here is characterized as follows: a) with "PTV" inlets temperature programmed; b) with gas line system of "Back Flushing"; c) with direct injection of oil samples. After oil sample injection, "Back Flushing" is on when light hydrocarbon components enter into analytical chromatographic column. At the same time, the temperature of inlet increases. The high temperature and "Back Flushing" blow the heavy components in the oil samples out of the analytical system. Besides, the analytical method of "Head Space" was established. Both "PTV with Back Flushing" and "Head Space" have the advantages of long column life and short analysis time. The resolution for lighter components < C9 meets the criterion of ASTM D5134-98, with the good repeatability. Ten oil samples from 6 oil areas were analysed by using the two methods. The relative deviations between the two analytical results represented by 19 geochemistry parameters were about +/- (1%-25%). The reasons for the deviation are discussed. It is pointed out that in geochemistry study it is not acceptable to combine the data obtained from two analytical methods. The analytical results obtained by injecting crude oil directly into injector are more reliable. The results obtained in "Head Space" analytical method should be calibrated when used in geochemistry study. PMID- 12545488 TI - [Advances in the fiber coating of solid phase micro-extraction]. AB - Solid phase micro-extraction (SPME) is a simple and effective sample preparation technique and fiber coatings play a very important role in SPME process. The discussion begins with a brief historical perspective from the very early work conducted almost a decade ago. At the moment, more than 20 alternatives of fiber coatings and size are available. In this paper, the coatings are divided into two categories: commercial fibers and custom-made fibers. A novel so-gel method has been introduced for the preparation of the coatings. PMID- 12545487 TI - [Capillary electrophoresis and chip capillary electrophoresis of carbohydrate]. AB - Carbohydrate has many important functions in various biologic processes. Capillary electrophoresis (CE) is one of the key tools of carbohydrate analysis. Chip CE is a new but efficient technique in the study of life science. Carbohydrate analysis with CE and Chip CE are reviewed. Separation strategies and detection of CE for various carbohydrates, including monosaccharides, polysaccharides and glycoconjugates are described. The application and potential ability of Chip CE in carbohydrate research are reviewed as well. PMID- 12545489 TI - [Prediction of retention parameters of explosives by artificial neural network]. AB - The quantitative relationship between the retention parameters and the structure of explosives is discussed. Molecular topological indices are used to represent the structure. Based on the back-propagation algorithm, a quantitative model was established after a training process of a train-set containing 20 explosives being completed. The Sigmoid function was chosen as the transmit function. The retention parameters (log kappa w and S) acted as output vectors, while molecular connecting indices (0 chi, 1 chi, 2 chi) and edge adjacent indices(epsilon) acted as input vectors. The input vectors were normalized in the range of -3-3 and the output vectors were normalized in the range of 0-1. The accuracy of network was 0.5 and the beginning value of studying pace (eta) was 0.2, the momentum factor (alpha) was 0.5. The results showed that the yield model reflected the relationship between the structure and retention index of compounds, and had high accuracy. Most of the relative errors were below +/- 5%. PMID- 12545490 TI - [Determination of lamotrigine in the serum and study on pharmacokinetics in rabbits by RP-HPLC]. AB - In the treatment of epileptic patients a method was developed to monitor the lamotrigine concentration by HPLC using a YWG-ODS column and a detector at 305 nm. The linear range was from 0.25 mg/L to 50 mg/L with a mean correlation coefficient of 0.9998. The average recovery was 93.78% and the standard deviation was 2.66%. Within and between-day precisions were 2.4% (n = 9) and 5.3% (n = 5) in RSD respectively. The pharmacokinetics was studied after a single dose of 25 mg/kg for 3 rabbits. Lamotrigine concentrations in serum were assayed and the parameters were computed with 3P87 program. The concentration-time curve of lamotrigine in rabbits fitted to a two compartment model, in which Cmax was 9.34 micrograms/mL, t1/2(Ke) was 8.78 h, and CL/f(s) was 0.69 L/h. The method is rapid, accurate and can be used as routine test in the use of lamotrigine. PMID- 12545491 TI - [Study of unified analytical method of compatibleness for traditional Chinese medicines. II. The quantitation of curve fitting]. AB - To introduce method of compatibleness analysis for traditional Chinese medicines, we select a part of the results from the study of astragalus root(A.), Chinese angelica root(Ca.) and their complex prescription (Danggui Buxue Tang, DBT) with an analytical mode of reversed-phase HPLC-photodiode array detector as an example of the quantitation of curve fitting. A new quantitative method, the concentration variation of components as the changing of single prescription dosage in complex prescriptions is discussed. This is the second part of the method of compatibleness analysis for traditional Chinese medicines. PMID- 12545492 TI - [Determination of unbound concentration of drug in drug-human serum albumin mixture by high performance frontal analysis]. AB - A high performance frontal analysis(HPFA) method was developed to determine the unbound concentration of drugs in drug-human serum albumin (HSA) mixture under binding equilibrium. The sample was injected directly onto an internal-surface reversed-phase silica column (ISRP). The mobile phase was 67 mmol/L phosphate buffer (pH 7.4, I = 0.17 mol/L). When a large volume of sample solution under drug-HSA binding equilibrium was directly injected, the drug was eluted as a trapezoidal peak with a plateau, and the drug concentration in this region was the same as that of the unbound drug in the sample solution. The eluate of plateau region was collected and a small volume was injected onto a reversed phase HPLC column. This HPFA-HPLC method was employed in the determination of unbound concentration in both ketoprofen (KP)-HSA and cefoperazone (CP)-HSA mixtures. The unbound concentrations of drugs obtained by using HPFA-HPLC were compared with those determined with ultrafiltration-HPLC. The effects of sample volume and flow rate of mobile phase on the plateau formation were investigated. It was found that the minimum injection volume to achieve a trapezoidal peak varied with drugs. The flow rate showed no effect on the trapezoidal peak formation. The unbound concentrations of KP and CP obtained were about the same by using HPFA-HPLC or ultrafiltration-HPLC and precisions were similar for both methods. PMID- 12545493 TI - [Determination of intestinal trefoil factor in burned rats by reversed-phase high performance liquid chromatography]. AB - A reversed-phase high performance liquid chromatographic method was established for determination of intestinal trefoil factor (ITF) in burned rats. The analysis was carried out on a Hypersil C4 column (30 nm, 5 microns, 4.6 mm i.d. x 250 mm) with gradient elution of acetonitrile-trifluoroacetic acid-water solution at a flow rate of 1 mL/min and detected by a UV detector at 214 nm. The volume proportion of trifluoroacetic acid in water is 0.1% and the volume proportion of acetonitrile in mobile phase ranged from 19.5% to 42.0%. The linear range of the method was 1 mg/L-100 mg/L with relative coefficient of 0.9989. The minimum detection limit was 0.5 mg/L. The recovery of ITF added ranged between 93.95% and 105.90%. The intra-day and inter-day RSD of ITF were less than 5.33% and 6.10% respectively. This method is precise, accurate and can be used for the determination of ITF in intestinal mucosal of burned rats. Therefore, it is helpful in the research of comparing the effects of enteral feeding and parenteral nutrition on ITF expression. PMID- 12545494 TI - [Analysis of new type composite antioxidant by HPLC/APCI-MS]. AB - An effective method for the analysis of a new type composite antioxidant has been developed. High performance liquid chromatography (HPLC) with diode-array detection (DAD) interfaced to atmospheric pressure chemical ionization-mass spectrometry (APCI-MS) was used. In this paper, the major components of the composite antioxidant additive were identified by means of HPLC/APCI-MS with positive/negative ion scan modes and programmable in-source collision induced dissociation and infrared(IR) spectra. The results show that this method is rapid, convenient and very useful for the analysis of lubricant additives. PMID- 12545495 TI - [Study on the determination of NO, AsH3, COS and other impurities in cracked gas by chromatography/mass spectrometry method]. AB - The selected ion monitoring(SIM) mode of capillary column gas chromatography coupled with mass spectrometry has been used for the determination of NO, AsH3, COS, thio-ether, thio-alcohol etc. in gases from catalytic cracking of heavy oils. The interference of CO, CO2, ethylene, ethane and N2 which affected the determination of NO was eliminated by chromatographic separation and subtraction of responses. The interference to the determination of selected ion was studied. Only C2H5SH was affected at m/z = 62 and the interference could be eliminated by using the ions at m/z = 47. The impurities in practical sample gas were determined. The detection limit of NO was 100 nL/L. The detection limits of other impurities were between 15 nL/L and 50 nL/L. PMID- 12545496 TI - [The determination of lorazepam in human urine by gas chromatography/nitrogen phosphorus detector]. AB - A method to assay lorazepam in human urine has been developed. After addition of hydroxyethylflurazepam (internal standard) and hydrolysis with beta glucuronidase, the lorazepam and hydroxyethylflurazepam were extracted with ethyl ether at pH 10.8. The analysis was performed on an HP-5 capillary column with nitrogen-phosphorus detector(NPD). The detection limit and recovery of analytes in urine were 5 micrograms/L and (83.4 +/- 3.1)% respectively. The method was successfully applied to urine specimens collected from healthy human volunteers who have ingested 2 mg of lorazepam. The method was sensitive enough to assay urine specimen excreted at 32 h after taking the medicine by volunteers. PMID- 12545497 TI - [Analysis of soybean lecithin by supercritical fluid chromatography]. AB - Separation of six phospholipids, phosphatidyl choline (PC), phosphatidyl ethanolamine (PE), phosphatidyl-serine(PS), phosphatidyl-inositol(PI), phosphotidic acid(PA), lyso-phosphatidyl-choline(lyso-PC), in soybean lecithin with supercritical fluid chromatography was achieved within 15 min. C18 column was used and carbon dioxide modified by ethanol containing 0.05% (V/V) triethylamine was chosen as the mobile phase. Effects of the composition of mobile phase, temperature and pressure were studied. The quantitative analysis of PC has been achieved with external standard method. The calibration curve for PC was linear in the range between 0.020 g/L-0.075 g/L and the detection limit was 0.2 microgram. This method has been applied to the analysis of PC in soybean lecithin. PMID- 12545498 TI - [Determination of active ingredients of 12 pesticides by capillary gas chromatography]. AB - A method for the determination of the active ingredients of 12 pesticides, including procymidone and tebuconazole, with capillary gas chromatography has been developed. A wide bore capillary column(SE-30) and FID detector were employed. The internal standards were used for quantitative determination and GC/MS for qualitative determination. The average recoveries were 99.20%-102.44% while the relative standard deviations were within the range of 0.07% to 5.49%. The method is simple, practical and accurate for the determination of the active ingredients of individual pesticide, formulation or multi-pesticides as a whole. PMID- 12545499 TI - [Analysis and separation of chromium(III) oligomers by ion chromatography]. AB - A method of analyzing chromium(III) oligomers in aqueous solution by means of TSK Gel ion exchange analytical column and diode array detector was developed. The effects of determination conditions, elution and ionic strength on analysis were investigated carefully and the analytical condition was optimized. The samples were separated for 10 min by using TSK-Gel SP-5PW(75 mm x 7.5 mm i.d., 10 microns) as column, 3 mol/L NaClO4-0.03 mol/L HClO4 as mobile phase, and detected at the wavelength of 200 nm. The method was simple, rapid and repeatable. This method was also used to determine the concentration of each fraction under various conditions of preparing chromium(III) oligomers and rendering the optimized preparation parameters. PMID- 12545500 TI - [Determination of organic and inorganic anions in snow and ice from alpine glaciers with DX-300 ion chromatograph]. AB - Organic and inorganic anions in snow and ice from Glacier 1 at Urumqi river head, Tianshan Mountain, Xinjiang, Northwest China, were determined using DIONEX-300 ion chromatograph with a lot of columns. AS4A-SC was selected as the separation column, AG4A-SC as the guard column, TAC-2 as the anion enrichment column, ATC-1 as the anion trap column and ASRS-II as the suppresser. It was conducted under gradient elution with sodium tetraborate (Na2B4O7) as eluent and 25 mmol/L H2SO4 as regeneration solution for chemical suppression. By preconcentration, 2 mL melted water of snow or ice can be analyzed for separation and quantitation of more than 10 anions, which were fluoride, acetate, formate, pyruvate, monochloroacetate, chloride, nitrite, bromide, nitrate, phosphate, sulfate and oxalate. The relative standard deviations for most of the anions were below 5%. PMID- 12545501 TI - [Determination of 4-chloro-3,5-dimethyl phenol in antiseptics and synthetic products by scanning thin-layer chromatography]. AB - A new method of scanning thin-layer chromatography was established for the quantitative determination of the 4-chloro-3,5-dimethyl phenol by developing with 40% benzene in trichloromethane on GF254 silica gel plate. The results showed that the linear dynamic range of 4-chloro-3, 5-dimethyl phenol was 0.1858 g/L 7.4320 g/L in which the relative peak area obeyed the Lambert-Beer's law with a correlation coefficient of 0.9994 and recoveries of 98.31%-100.1%. The method is simple, rapid, sensitive and reproducible. PMID- 12545502 TI - [Determination of Henry's law constants for dimethyl sulfide in seawater]. AB - The article presents the method of determination of Henry's Law Constant(K) for dimethyl sulfide (DMS) in seawater using multiple phase equilibration/headspace gas chromatography. The method is based on the demonstration that analysis of only one phase after multiple phase equilibration give all necessary data. The values of K of DMS in seawater determined by this method was 0.089 with relative standard deviation(RSD) of 5.50% at 22 degrees C. The linear relationship between log K and w (NaCl) was log K = 0.0688.w (NaCl) - 1.3568 with correlation coefficient(r) of 0.997. The regression equation of log K against reciprocal of absolute temperature(T) was log K = -1,544.8/T + 4.2112 with r of 0.996. PMID- 12545503 TI - [Capillary array electrophoresis and high-throughput DNA sequencing]. AB - Depending on the outputs of 10,080 DNA sequencing samples from the human genome project, technical features of capillary array electrophoresis are discussed. The results show that 88% of the total readlength were higher than 50 kbp/run. It means that most of the readlength could reach 500 bp or more with reasonable quality. The results were much better than expected when the technology of capillary array appeared about one year ago. A key parameter affecting the quality of DNA sequencing is the purity of sequencing template. Therefore, a batch agarose slab electrophoresis was applied to check the purity before loading. Other conditions, such as concentration of template and loading amount, are discussed as well. Finally, comparisons based on the electrophoresis timing, the pre-treatment of samples, the loading procedures and the results analysis between capillary array electrophoresis and slab gel electrophoresis have been made. From the point of view of high-throughput DNA sequencing, especially in the field of genome research, the capillary array electrophoresis should be the better choice as a more efficient technical platform. PMID- 12545504 TI - [Quantitative determination of icariin in "tangzhi shuangjiang cha" by high performance liquid chromatography]. AB - Icariin in "Tangzhi Shuangjiang Cha" was quantitatively determined by HPLC with ODS column, a mixture of MeOH-H2O(60:40, V/V) as mobile phase and UV detection at 270 nm. There was a good linear relationship within the range of 0.1 g/L-0.5 g/L(r = 0.9993). The average recovery was 101.4%, RSD was below 2.7%(n = 3). The method is rapid, simple, accurate and reproducible. And the method is proposed to be used for quality control of "Tangzhi Shuangjiang Cha". PMID- 12545505 TI - [Reversed-phase high performance liquid chromatography used for determination of amino acids of parasites]. AB - The compositions and contents of amino acids of parasites in different areas of China have been determined by precolumn derivatization, reversed-phase high performance liquid chromatography (HPLC) separation and ultra-violet absorption quantitation in order to prevent diseases caused from parasites. The results show that there were some differences in contents of amino acids in B. malayi from seven different areas in China. The contents of amino acids gradually increased from the southern to the northern areas in China. The method is simple and accurate, and the results have important values to investigate the life activities of parasites. PMID- 12545506 TI - [Determination of salicylic acid, resorcinol and chloramphenicol in junlieping tincture by reversed-phase high performance liquid chromatography]. AB - A reversed-phase high performance liquid chromatographic method was developed for the quantitative determination of salicylic acid, resorcinol and chloramphenicol in Junlieping tincture. The operation was carried out on an ODS column with the mobile phase of a mixture of acetonitrile-methanol-water(25:10:65, volume proportion). The eluate was detected at 278 nm wavelength. This method is simple, fast and can be used for the quality control of Junlieping tincture. PMID- 12545507 TI - [Determination of isophthalic acid in polyethylene glycol terephthalate fiber by high performance liquid chromatography]. AB - The isophthalic acid(IPA) is present in polyethylene glycol terephthalate(PET) at small level, but plays an important role for the properties of polyester fiber. Thus the PET sample was hydrolysated by 100 g/L of sodium hydrate in methanol at 70 degrees C during 10 h and finally assayed by HPLC. A reversed-phase high performance liquid chromatographic method for the separation of isophthalic acid from terephthalic acid(TPA) has been developed. The operating conditions were Novapak C18 column, methanol-water(15:85, V/V)(pH3) as mobile phase with the flow rate of 1.0 mL/min and UV detection wavelength at 254 nm. PMID- 12545508 TI - [Determination of organic acids in cigarette smoke]. AB - Solid phase microextraction (SPME) is a simple, cost-effective, time-saving extraction technique. It is rapid and highly sensitive. In this study, organic acids in cigarette smoke were extracted with 85 microns polyacrylate coated fiber. Volatile and semi-volatile organic acids, such as butanoic acid, hexanoic acid, 2-furancarboxylic acid, octanoic acid, nonanoic acid, benzoic acid, phenylacetic acid, tetradecanoic acid, and hexadecanoic acid from cigarette smoke were converted to their esters by using methanolic derivatization agent. The esters were extracted by using headspace solid phase microextraction and determined quantitatively by gas chromatography with selected ion monitoring mode mass spectrometry. Qualitative analysis of 4 brands cigarette smoke demonstrated the usefulness of this analytical technique of volatile and semi-volatile acids in cigarette smoke. More than sixty organic acids were detected by using this method. PMID- 12545509 TI - [Determination of ammonia and potassium in tobacco by ion chromatography]. AB - A new ion chromatographic method was developed for the quantitative determination of ammonia and potassium in tobacco. The tobacco samples were treated with dilute nitric acid and K+, NH4+ were extracted. The method is fast and convenient. The relationships between peak heights and initial concentrations of K+ and NH4+ were linear. The detection limits were 0.05 mg/L for K+ and 0.03 mg/L for NH4+. Recoveries of K+ and NH4+ were 96.7%-106.7% and 96.5%-109.1% respectively. The whole procedure of determination can be completed within 5 min. PMID- 12545510 TI - [Determination of inorganic anions in beer and brewing water by ion chromatography]. AB - An ion chromatography method was used to determine fluoride, chloride, nitrite, dihydrogen phosphate, nitrate and sulfate anions in beer and brewing water on a YSA8 8022A-9(250 mm x 4 mm i.d.) separation column, with sodium carbonate-sodium bicarbonate buffer as mobile phase and an electrical conductivity cell as detector. Qualitative analysis was performed with retention time and stanard spiking method, and quantitative analysis was conducted with a calibration curve by measuring the peak height. This method is simple, rapid and accurate. PMID- 12545511 TI - [Analysis of writing age of ballpoint pen ink by thin-layer chromatography]. AB - A method based on using a thin-layer chromatographic method (TLC) for determining changes in dye of ballpoint pen ink developed with writing age. An ink drawn line of 1 cm in length was cut and placed in Eppendorf tube. Extraction was performed with 20 microL ethanol at room temperature for 15 min, and heated in steambath for 10 min. The ethanol solution(4 microL) containing the extracted dyes was then spotted on TLC plate by syringe after cooling. The ballpoint pen ink spots were eluted in a solvent of isopropyl alcohol. Using TLC scanner, the spots on TLC plate were scanned at 580 nm. The relations between ratio of peak heights and writing age were attained. Thus, the information on relative writing age can be obtained. And, the powerful evidence for identifying the age of ballpoint pen ink is provided by this method in the field of forensic science. PMID- 12545512 TI - Virtual ambulatory care. Computer simulation applications. AB - Computer simulation modeling has evolved during the past twenty years into an effective tool for analyzing and planning ambulatory care facilities. This article explains the use of this tool in three case-study, ambulatory care settings--a GI lab, holding beds for a cardiac catheterization laboratory, and in emergency services. These examples also illustrate the use of three software packages currently available: MedModel, Simul8, and WITNESS. PMID- 12545513 TI - Primary health care teams. Opportunities and challenges in evaluation of service delivery innovations. AB - Quality of service and corporate productivity may be improved when employees are formed into semi-autonomous teams that are empowered to be innovative in service and are rewarded for attaining performance goals. This article describes the planned staffing and implementation strategies for three models of primary health care teams (HCTs) in a managed care organization (MCO). Implementation of each model focussed on (1) changes to primary care staffing mix (structural integration of roles) and (2) development of teamwork (functional integration of roles). Evidence from other industries suggests that the planned changes to structural and functional integration of existing primary care delivery models could improve system productivity, patient satisfaction, clinical quality, and employee morale. Retrospective evaluations of whether the planned improvements were achieved, and whether these achievements can be attributed to changes in staffing mix or teamwork, are now being conducted. Opportunities and limitations in conducting these retrospective evaluations are discussed, particularly with reference to use of existing data sources. The article concludes with recommendations for evaluation of natural experiments in primary care redesign. PMID- 12545514 TI - Financial incentives and physicians' perceptions of conflict of interest and ability to arrange medically necessary services. AB - Many believe that physician payment mechanisms that include incentives to restrain utilization create conflicts of interest for physicians and result in the withholding of needed services. Pooled data from two rounds of the Community Tracking Study physician survey, a nationally representative telephone survey of physicians, are the basis of this analysis. We examine the association between explicit financial incentives linked to physician profiling and perceived conflict of interest, and the reported ability to obtain specific, medically necessary secondary services (referrals, hospitalizations and diagnostic imaging). Logistic regression models were employed to control for potentially confounding influences. After controlling for other factors, physicians subject to profiling linked with financial incentives were much less likely than physicians not affected by profiling to strongly agree that they can make clinical decisions in the best interests of their patients without the possibility of reducing their income. They were also less likely to report that they could always obtain selected medically necessary secondary services for their patients. Physicians subject to explicit financial incentives based on profiling are more likely than other physicians to perceive a conflict of interest. Physicians with financial incentives tied to profiling also experience greater difficulty obtaining medically necessary secondary services for their patients. PMID- 12545515 TI - Enrolling uninsured children in SCHIP. Lessons learned from community health centers. AB - In 1997 Congress established the State Child Health Insurance Program (SCHIP) to address the problem of the nation's low-income uninsured children. To help children become eligible for SCHIP and to tap into the potential revenue stream for previously uninsured children, community health centers have taken differing approaches. This study examines the lessons learned from enrolling children at 14 health centers in six states. The lessons can be valuable for primary care centers and other safety-net providers. PMID- 12545516 TI - Ambulatory patient safety. What we know and need to know. AB - While safety risks are widespread in ambulatory settings, there has been insufficient attention directed at developing the evidence base that is needed to improve ambulatory safety. In this article, the current state of knowledge about ambulatory safety is reviewed. A research agenda in ambulatory safety is proposed, as well as a series of potential interventions that could be used to improve safety in the ambulatory setting. PMID- 12545517 TI - Knowledge and opinions about clinical research. A cross-sectional survey in a sample of Italian citizens. AB - Large samples of the general population are exposed to the possibility of being enrolled in clinical trials on preventive strategies. Because little information is available on the general population, data were collected on what Italians know and how they feel about clinical research and factors that might help implement measures to improve participation in the main phases of clinical research were assessed. A random sample of 950 citizens representative of the Italian population was interviewed by ad hoc trained interviewers, using a standardized questionnaire with seven multiple-choice questions and two scenarios. Participants' knowledge and opinions about clinical research were in general limited, attitudes toward participation in clinical studies were unsatisfactory, and the results varied significantly according to sociodemographic and geographic variables. These results give a description of the people's level of knowledge and attitude about clinical research in Italy and also clarify the importance of factors that may be related to the study's principal variables. These findings might be useful in helping design and implement effective interventions to improve people's participation in the main phases of clinical research. PMID- 12545518 TI - Understanding attitudes toward clinical research. AB - General attitudes toward clinical research can have substantial effects on the research process. Recent studies, including the study by Apolone and Mosconi in this issue, suggest that certain attitudes toward research and evaluations of the risks and benefits of therapies may differ according to various cultural, rhetorical, and health contexts. Researchers must work to further understand societal attitudes toward research and to improve the attractiveness of clinical research for the general population. PMID- 12545519 TI - Juvenile justice. A role for health professionals. AB - The United States is the only nation in the world that continues to execute its youth. The use of the death penalty against those who committed crimes as children is an act contrary to American standards of decency and fairness, as well as international law. The adolescent brain has not fully developed before the age of 18 years of age. Thus children do not have the same emotional and mental capacity as adults. Although juveniles should be held accountable for their crimes, the United States must not impose this most extreme punishment. The medical profession must take a stand to stop the execution of juvenile offenders in the United States. PMID- 12545520 TI - Smallpox, big worries. Preparing medical-response teams is easier said than done, according to healthcare providers across the nation. AB - Preparations for healthcare workers to respond to a smallpox attack pose an array of daunting challenges, both financial and logistical, that must be addressed in a few weeks. By the end of the month, a massive corps of doctors, nurses and other healthcare workers will voluntarily receive the smallpox vaccine as part of the first phase of a national strategy announced last month by President Bush. PMID- 12545521 TI - West Virginia docs walk off the job in malpractice dispute. Protest on New Year's Day shows rising frustration with high cost of malpractice insurance coverage. PMID- 12545522 TI - Expectations high over Frist. PMID- 12545523 TI - Going to the pols. San Antonio health system pushes tax plan to help fund trauma centers in Texas. PMID- 12545524 TI - Time to settle up. Hospitals pay $12.3 million but admit no wrongdoing in year end settlements. PMID- 12545525 TI - The next wave in payment looms. Consumer-driven health plans confer new risks on patients and providers. PMID- 12545526 TI - Outlook '03. Even with unrelenting cost increases, some sectors enjoy a healthier prognosis. But the year promises formidable challenges for all. PMID- 12545527 TI - Defender of the law. False Claims Act co-author to keep fighting healthcare fraud as Senate panel chair. PMID- 12545528 TI - GHX and Medibuy merge, create e-commerce giant. PMID- 12545529 TI - Price survey. Needle market in delicate balance. PMID- 12545530 TI - Firm has life, soul and Internet presence. PMID- 12545531 TI - Cardiology prices seen levelling off in '03. AB - The price outlook for nearly 200 product categories, complied by Consorta, is featured this month. PMID- 12545532 TI - Quantitative estimation of viral fitness using Pyrosequencing. PMID- 12545533 TI - Selection of hygromycin-resistant Arabidopsis seedlings. PMID- 12545534 TI - Rapid genomic identification of transgenic integration sites for mouse insertional mutants. PMID- 12545535 TI - Deep screening of recombination proficient bacteriophage libraries. PMID- 12545536 TI - Laser capture microdissection of paraffin-embedded tissues. PMID- 12545537 TI - Isolation of intact RNA following cryosection of archived frozen tissue. PMID- 12545538 TI - Megaprimer mutagenesis using very long primers. PMID- 12545539 TI - Reverse-polarity PAGE for examining DNA binding domain phosphorylation. PMID- 12545540 TI - Plastic contaminant masquerades as DNA in mutation detection by denaturing HPLC. PMID- 12545541 TI - Silver particles enhance emission of fluorescent DNA oligomers. AB - Here we describe a new opportunity in methodology for increasing the detectability of fluorescently labeled DNA on solid substrates. We show that the use of glass substrates coated with metallic silver particles results in an approximate 5-fold increase in the intensity of Cy3- or Cy5-labeled DNA oligomers. Proximity to these silver particles also increases the photostability of Cy3- and Cy5-labeled oligomers. These results suggest the use of DNA array substrates with silver particles for increased sensitivity in genetic analysis. PMID- 12545542 TI - Molecular beacon sequence design algorithm. AB - A method based on Web-based tools is presented to design optimally functioning molecular beacons. Molecular beacons, fluorogenic hybridization probes, are a powerful tool for the rapid and specific detection of a particular nucleic acid sequence. However, their synthesis costs can be considerable. Since molecular beacon performance is based on its sequence, it is imperative to rationally design an optimal sequence before synthesis. The algorithm presented here uses simple Microsoft Excel formulas and macros to rank candidate sequences. This analysis is carried out using mfold structural predictions along with other free Web-based tools. For smaller laboratories where molecular beacons are not the focus of research, the public domain algorithm described here may be usefully employed to aid in molecular beacon design. PMID- 12545543 TI - Creating precise GFP fusions in plasmids using yeast homologous recombination. AB - Using a combination of primer amplification, homologous recombination, and yeast genetics, we established a method for creating precise promoter and protein fusions in genes originating from organisms other than yeast. One major advantage of this new method is its versatility. Fusions can be produced within a target gene without constraints regarding the site of insertion. Thus, fusions can be generated within a target sequence exactly at the site desired, and all sequences upstream and downstream of the insertion site were preserved. To illustrate the general applicability of this technique, we fused the gene encoding GFP to a Caenorhabditis elegans homologue of the dishevelled gene, dsh-2. This approach is not restricted to GFP fusions but can be utilized to create fusions between almost any two sequences regardless of the source. Therefore, this method provides a flexible alternative to other PCR-mediated techniques. PMID- 12545544 TI - Baculovirus expression vectors that incorporate the foreign protein into viral occlusion bodies. AB - Current baculovirus expression systems typically produce soluble proteins that accumulate within the infected insect cell or are secreted into the growth medium. A system has now been developed for the incorporation of foreign proteins, along with the matrix protein, polyhedrin, into baculovirus occlusion bodies. Initial studies showed that a recombinant virus expressing a translational fusion between polyhedrin and GFP did not form occlusion bodies. However, a baculovirus coexpressing native polyhedrin and the polyhedrin-GFP fusion protein formed occlusion bodies that fluoresced under UV light, demonstrating that they included the polyhedrin-GFP fusion protein. This was confirmed by immunoblot analysis. Thus, incorporation of a foreign protein into occlusion bodies depends on an interaction between native polyhedrin and the polyhedrin fusion protein. Electron microscopy demonstrated that the occlusion bodies containing GFP also incorporated virions as expected. These ColorPol occlusion bodies were as infectious to insect larvae as occlusion bodies produced by wild-type virus. This new system expands the capabilities for foreign gene expression by baculoviruses, which has implications for biopesticide design, novel vaccine delivery systems, and fusion protein purification applications. PMID- 12545545 TI - Exogenous reference RNA for normalization of real-time quantitative PCR. AB - We have utilized an in vitro transcribed 3' mRNA fragment of the plant gene ribulose bisphosphate carboxylase (RuBisCO) as an exogenous standard for normalization of quantitative PCR data. Both K562 cells and primary erythroid CD34+ progenitor cells were treated with sodium butyrate and changes in gamma globin mRNA levels were assayed using a previously published TaqMan probe and primer set, while RuBisCO levels were assayed by a SYBR Green detection assay. The data presented show that a correction to measured gamma-globin induction was necessary with both cell types. The correction for the CD34+ progenitor cells was a striking 95% increase, while that for the K562 cells was 44%. The use of an exogenous reference such as in vitro transcribed mRNA for the RuBisCO plant gene provides a robust and sample-independent method for the normalization of quantitative PCR data in bacterial and animal cells. PMID- 12545546 TI - Extraction of PCR-quality plant and microbial DNA from total rumen contents. AB - DNA from rumen digesta has several diagnostic applications such as studying microbial community dynamics, transgene/DNA stability, and population typing of various rumen bacteria. Several DNA extraction procedures are described in the literature for rumen digesta, which describe the removal of tannins, polysaccharides, and other PCR inhibitors. Some of these protocols are time consuming and impractical when handling a large number of samples routinely. Here we describe a rapid method for the extraction of PCR-quality plant and microbial DNA from total rumen contents that is based on modifications in the cetyltrimethylammonium bromide procedure followed by cleanup using a Qiagen column. This procedure is highly reproducible and relatively short, once the initial grinding of the samples is performed, and it consistently yields PCR quality DNA. PMID- 12545547 TI - High-sensitivity quantitative PCR platform. AB - Real-time PCR methods have become widely used within the past few years. However, real-time PCR is rarely used to study chronic diseases with low pathogen loads, presumably because of insufficient sensitivity. In this report, we developed an integrated nucleic acid isolation and real-time PCR platform that vastly improved the sensitivity of the quantitative detection of the intracellular bacterium, Chlamydia spp., by fluorescence resonance energy transfer real-time PCR. Determinants of the overall detection sensitivity were analyzed by extracting nucleic acids from bovine milk specimens spiked with low amounts of chlamydial organisms. Nucleic acids were optimally preserved and recovered by collection in guanidinium stabilization buffer, binding to a matrix of glass fiber fleece, and elution in low volume. Step-down thermal cycling and an excess of hot-start Taq polymerase vastly improved the robustness and sensitivity of the real-time PCR while essentially maintaining 100% specificity. The amplification of Chlamydia 23S rRNA allowed for the differentiation of chlamydial species and was more robust at low target numbers than amplification of the omp1 gene. The best combined method detected single targets per a 100-microL specimen equivalent in a 5-microL real-time PCR input. In an initial application, this high-sensitivity real-time PCR platform demonstrated a high prevalence of chlamydial infection in cattle. PMID- 12545548 TI - Electrophoretic signal comparison applied to mRNA differential display analysis. AB - Gene expression analysis by electrophoretic methods is currently limited by the labor-intensive visual evaluation of the electrophoretic signal profiles. For this purpose, we present a flexible approach to computer-assisted comparison of quantitative electrophoretic patterns between multiple expression signals. Gaussian curves are first fitted to the complex peak mixtures, and the resulting approximate signals are then aligned and compared on a peak-by-peak basis with respect to specific patterns defined by the investigator. The rationale of the method is to produce a compressed list of exceptional expression patterns quantified by a set of associated numeric features. A score value is attached to each pattern in such a way that large values identify the most potential findings to be focused on in visual analysis instead of the vast amount of original electrophoretic results. The validity of the method is demonstrated by analyzing a large set of electrophoretic data from mRNA differential display experiments monitoring changes in gene expression patterns in human colonic carcinoma. The automated identification of variously defined gene expression patterns agrees well with the visual evaluation of the same electropherograms. The general comparison approach may also be found useful with other gene expression profiling instruments. PMID- 12545549 TI - Microarray of recombinant antibodies using a streptavidin sensor surface self assembled onto a gold layer. AB - We have developed a sensitive method for the detection of recombinant antibody antigen interactions in a microarray format. The biochip sensor platform used in this study is based on an oriented streptavidin monolayer that provides a biological interface with well-defined surface architecture that dramatically reduces nonspecific binding interactions. All the antibody or antigen probes were biotinylated and coupled onto streptavidin-coated biochip surfaces (1 microL total volume). The detection limits for the immobilized probes on the microarray surface were 0.5 microgram/mL (200 fmol/spot) for the peptide antigen and 0.1 microgram/mL (3 fmol/spot) for the recombinant antibodies. Optimal concentrations for the detection of the Cy5-labeled protein target were in the range of 20 micrograms/mL. Protein microchips were used to measure antibody-antigen kinetics, to find optimal temperature conditions, and to establish the shelf life of recombinant antibodies immobilized on the streptavidin surface. For recombinant antibody fragments with a kDa of 10-100 nM, we have established an easy and direct immunoassay. In addition, we developed an indirect method for antibody detection with no need for expensive and time-consuming antibody purifications and modifications. Such a method was shown to be useful for large-scale screening of recombinant antibody fragments directly after their functional expression in bacteria. Our data demonstrate that recombinant antibody fragments are suitable components in the construction of antibody chips. PMID- 12545550 TI - QSYP peptide sequence is selected from phage display libraries by bovine IgG contaminants in monoclonal antibody preparations. AB - A consensus peptide sequence, QSYP, appears as an artifact during the mapping of monoclonal antibodies (MAbs) using a random peptide phage display library. Phage bearing this QSYP sequence were independently selected by four different laboratories screening separate MAb preparations with the same phage library. In each case, the QSYP sequence was selected in addition to a consensus sequence specific to the MAb. Phage that displayed the QSYP sequence were not bound by the MAb of interest, but rather bound to bovine IgG derived from the FBS present in the hybridoma growth media. The implications of this finding for the interpretation of phage library screening results and possible methods for the removal of bovine IgG from MAb preparations are discussed. PMID- 12545551 TI - Transient transfection of the human myeloid cell line Mono Mac 6 using electroporation. AB - Leukemic cell lines such as Mono Mac 6 provide an excellent model for studying changes in gene expression during induction of cell differentiation. Mono Mac 6 cells can be induced to differentiate from their immature state to cells resembling morphologically and functionally mature monocytes and macrophages by various stimuli such as calcitriol and transforming growth factor-beta. During differentiation, the expression of differentiation markers such as the cell surface antigen CD14 or other differentiation-related genes such as 5 lipoxygenase are strongly increased. Thus, this cell line constitutes an excellent model system to study the regulation of gene expression by inducers of cell differentiation. However, myeloid cell lines are often refractory to transfection by calcium phosphate or DEAE dextran so that reporter gene assays are difficult to perform. We have established a transient transfection protocol for Mono Mac 6 cells using electroporation, a 5-lipoxygenase promoter luciferase reporter gene construct, and the secreted alkaline phosphatase as an internal standard. PMID- 12545552 TI - Adeno-associated virus vectors for therapeutic gene transfer. PMID- 12545553 TI - Tracing the origin of non-hematopoietic cells using CD45 PCR restriction fragment length polymorphisms. PMID- 12545554 TI - No-wash dyes for calcium flux measurement. PMID- 12545555 TI - Expression of ssDNA in mammalian cells. AB - Antisense therapy involves the use of antisense oligonucleotides for altering targeted gene function. However, the low efficiency of cell delivery of antisense oligonucleotides has limited the efficacy of antisense therapeutic approaches. RNA-based antisense or ribozyme oligonucleotides can be either synthesized endogenously (e.g., by a viral vector) or delivered exogenously. However, there is presently no vector delivery system available for DNA-based oligonucleotides. Recently, a novel ssDNA expression vector that can generate intracellularly any ssDNA molecule, such as antisense oligonucleotide or DNA enzyme, has been developed in our laboratory. Here we describe an improved expression vector based on the first-generation two-vector system. To test this new expression vector, we chose to express a single-stranded "10-23" DNA enzyme targeting c-raf mRNA in the human lung carcinoma A549 cell line. After introduction into cells by transient transfection, c-raf-cleaving DNA enzymes produced by this expression vector can significantly suppress the expression of c-raf mRNA. Furthermore, the expressed c raf DNA enzymes induced cell apoptosis, as indicated by genomic DNA fragmentation assay. Our study further demonstrates the feasibility of using this novel ssDNA expression technology to produce intracellularly any sequence of interest, including antisense oligonucleotides and DNA enzyme molecules. PMID- 12545556 TI - Quantitative detection of low-copy-number mRNAs differing at single nucleotide positions. AB - Accurate analysis of mRNA expression levels of SNPs, highly homologous genes, and splicing variants requires techniques capable of quantifying low-copy-number mRNAs differing at single nucleotide positions. We have used an RT-PCR-based technique based on co-amplification of closely related target mRNA transcripts and assessed the effect of the stochastic distribution of low-copy-number templates on sampling variation when quantifying rare mRNA transcripts. The technique was optimized for maximal sensitivity to enable the analysis of samples containing a subpopulation of target cells and small microdissected samples. We demonstrate that the input level of template molecules is a critical determinant of the achievable assay precision. A minimum of approximately 50 molecules of template is required to discriminate between 2-fold differences in the expression levels of two transcripts. At levels above 1000 molecules of input template, the stochastic effects on sampling variation become negligible. PMID- 12545557 TI - Use of adenovirus vectors for functional gene analysis in the chicken chorioallantoic membrane. AB - The chicken chorioallantoic membrane (CAM) assay represents one of the most widely used in vivo screening assay for genes with angiogenic (blood vessel inducing) or angiostatic (inhibition of vessel formation or their destruction) activities. Here we show that adenovirus gene transfer vectors infect cells in the CAM and lead to expression of the viral transgene. Furthermore, infection with an adenovirus vector containing the human vascular endothelial growth factor gene induced the formation of new blood vessels. This improved method saves a considerable amount of time in the identification of genes that can influence blood vessel formation because the expensive and time-consuming production and purification of recombinant protein can be omitted. PMID- 12545558 TI - Optimized production of high-titer recombinant adeno-associated virus in roller bottles. AB - Adeno-associated viral (AAV) vectors are used for in vivo gene transfer in a number of preclinical models of genetic diseases (including large-animal models) and are currently being tested in clinical trials for treatment of hemophilia B and cystic fibrosis. Protocols for production of AAV vectors in a helper virus free system are available and are based on transient transfection of HEK-293 cells with multiple plasmids. Scale-up of vector production has been labor intensive and inefficient because of a lack of larger culture vessels suitable for growth of adherent cells, large-scale transfection, and vector production. Here we report efficient production of AAV vector in roller bottles, which represents a 10-fold scale-up from the conventional flask or plate method. Optimized production yielded greater than 10(13) vector genomes per bottle and was as cost effective as published protocols using plates. Successful vector production by this method was dependent on optimization of transfection by calcium phosphate precipitation, of monitoring of cell growth (by measurement of glucose consumption), of cell culture conditions, and CO2/air exchange with the culture vessel. PMID- 12545559 TI - Kinetic FP-TDI assay for SNP allele frequency determination. AB - Strategies for identifying genetic risk factors in complex diseases by association studies require the comparison of allele frequencies of numerous SNPs between affected and control populations. Theoretically, hundreds of thousands of SNP markers across the genome will have to be genotyped in these studies. Genotyping SNPs one sample at a time is extremely costly and time consuming. To streamline whole genome association studies, some have proposed to screen SNPs by pooling the DNA samples initially for allele frequency determination and perform individual genotyping only when there is a significant discrepancy in allele frequencies between the affected and control populations. Here we describe a new method for determining the allele frequency of SNPs in pooled DNA samples using a two-color primer extension assay with real-time monitoring of fluorescence polarization (named kinetic FP-TDI assay). By comparing the ratio of the rate of incorporation of the two allele-specific dye-terminators, one can calculate the relative amounts of each allele in the pooled sample. The accuracy of allele frequency determination with pooled samples is within 3.3 +/- 0.8% of that determined by genotyping individual samples that make up the pool. PMID- 12545560 TI - Improved sensitivity for solid-support invasive cleavage reactions with flow cytometry analysis. AB - A new configuration of the solid-support invasive cleavage reaction provides a small reaction-volume format for high-sensitivity discrimination of nucleic acid targets with single nucleotide differences. With target concentrations as low as 2 amol/assay, the solid-support invasive cleavage reaction clearly distinguishes single base mutations. Two oligonucleotides tethered to the solid support hybridize to the target nucleic acid, forming a tripartite substrate that can be recognized and cleaved by Cleavase, a structure-specific 5'-nuclease. Each cleavage event yields fluorescence signal on the surface. When microspheres serve as the solid-support surface, analysis by fluorometer imparts real-time information about change in the reaction signal over time. Flow cytometry provides an alternative detection technology that collects endpoint information about the reaction signal on individual microspheres. A reaction volume of 10 microL with as few as 3000 microspheres is sufficient to distinguish single nucleotide differences at target concentrations less than 200 fM. This sensitivity level is within the range required for analysis of SNPs in genomic DNA. In addition, the flow cytometry format has multiplexing potential, making the microsphere-based invasive cleavage assay attractive for high-throughput genomic applications. PMID- 12545561 TI - DNA sequencing using rolling circle amplification and precision glass syringes in a high-throughput liquid handling system. AB - An automated high-throughput method that employs rolling circle amplification (RCA) to generate template for large-scale DNA sequencing has been developed using liquid handling systems equipped with precision glass syringes. A protocol was designed to perform the sequencing analysis from template preparation to thermal cycle sequencing within the same vessel, thus minimizing the amount of liquid handling and transfer. The amplified DNA was directly used for cycle sequencing with no need for any purification procedures. Total RCA reaction volumes as low as 500 nL generated sufficient templates for successful sequencing. Reducing the RCA total reaction volumes by a 40-fold factor, from a total of 20 microL to 500 nL, resulted in a significant reduction in cost, from $1.25/reaction to less than $0.04/reaction. Additionally, the volume of the sequencing reactions was reduced from a total of 20 to 10 microL, thus generating a further cost advantage. This high-throughput DNA sequencing protocol maximizes the speed and precision of processing while significantly reducing the cost of amplification. PMID- 12545562 TI - Your mind your body. PMID- 12545563 TI - The power of mood. PMID- 12545564 TI - Mother nature's little helpers. PMID- 12545565 TI - Say "om" before surgery. PMID- 12545567 TI - Can Freud get his job back. PMID- 12545566 TI - Is there a formula for joy? PMID- 12545568 TI - If everyone were on Prozac.... PMID- 12545569 TI - Through the ages. PMID- 12545570 TI - Masters of denial. PMID- 12545571 TI - What's sex got to do with it? PMID- 12545572 TI - One family's burden. PMID- 12545574 TI - Are your genes to blame? PMID- 12545573 TI - Postcards from the brain. PMID- 12545575 TI - The search for a murder gene. PMID- 12545576 TI - How we get labeled. PMID- 12545577 TI - Let's not overanalyze this. PMID- 12545578 TI - Your A to Z guide to the year in medicine. PMID- 12545579 TI - Pow! How to get it right in the kisser. PMID- 12545580 TI - Viagra turns 5. PMID- 12545581 TI - Illinois court wrestles with consent issue. Curtis v. Jaskey. PMID- 12545582 TI - On-call physician responsibility examined by Indiana court. Miller v. Martig. PMID- 12545583 TI - Flawed credentialing procedures create potential for hospital liability. Megrelishvili v. Our Lady of Mercy Medical Center. PMID- 12545584 TI - Health promoting community design: a new paradigm to promote healthy and active communities. AB - Richard Killingsworth explains that we have engineered routine forms of physical activity out of our daily lives, and that this is an important contributor to the obesity epidemic which claims at least 200,000 lives each year in the United States alone. A new field of study is emerging to address this problem, and the American Journal of Health Promotion has launched a new editorial section called "Health Promoting Community Design" to provide a forum to disseminate research in this new area. PMID- 12545585 TI - Evaluation of a community-based intervention to promote physical activity in youth: lessons from Active Winners. AB - PURPOSE: To test the effects of a community-based physical activity intervention designed to increase physical activity and to conduct an extensive process evaluation of the intervention. DESIGN: Quasi-experimental. SETTING: Two rural communities in South Carolina. One community received the intervention, and the other served as the comparison. SUBJECTS: Public school students who were in fifth grade at the start of the study (558 at baseline) were eligible to participate. A total of 436 students participated over the course of the study. INTERVENTION: The intervention included after-school and summer physical activity programs and home, school, and community components designed to increase physical activity in youth. The intervention took place over an 18-month period. MEASURES: Students reported after-school physical activity at three data collection points (prior to, during, and following the intervention) using the Previous Day Physical Activity Recall (PDPAR). They also completed a questionnaire designed to measure hypothesized psychosocial and environmental determinants of physical activity behavior. The process evaluation used meeting records, documentation of program activities, interviews, focus groups, and heart rate monitoring to evaluate the planning and implementation of the intervention. RESULTS: There were no significant differences in the physical activity variables and few significant differences in the psychosocial variables between the intervention and comparison groups. The process evaluation indicated that the after-school and summer physical activity component of the intervention was implemented as planned, but because of resource and time limitations, the home, school, and community components were not implemented as planned. CONCLUSIONS: The intervention did not have a significant effect on physical activity in the target population of children in the intervention community. This outcome is similar to that reported in other studies of community-based physical activity intervention. PMID- 12545586 TI - The relationship between National Heart, Lung, and Blood Institute Weight Guidelines and concurrent medical costs in a manufacturing population. AB - PURPOSE: To explore the relationship between the 1998 National Heart, Lung, and Blood Institute (NHLBI) weight guidelines and concurrent medical costs. DESIGN: Cross-sectional study. SETTING: In a nationwide manufacturing corporation (General Motors Corporation). SUBJECTS: A total of 177,971 employees, retirees, and their adult dependents who were enrolled in Indemnity/PPO health insurance plan during the years 1996 and 1997 and completed one health risk appraisal (HRA) in the same period. MEASURES: The participants were categorized into six weight groups according to the NHLBI 1998 guidelines (body mass index [BMI] < 18.5, 18.5 24.9, 25-29.9, 30-34.9, 35-39.9, > or = 40 kg/m2). The height and weight data were collected by self-reported values on an HRA or biometric screening completed during 1996 to 1997. To represent the typical medical costs in a given group, the median, instead of mean, medical charges were used in this article. The annual median medical charges (including drug charges) for years 1996 and 1997 were compared among the six weight groups by using Wilcoxon rank sum tests. The differences in median charges were also tested between the normal weight group and the other five groups for each of the 10 gender-age subgroups (five age groups: 19-44, 45-54, 55-64, 65-74, 75+). RESULTS: Overall median medical costs were consistent with the NHLBI weight guidelines. The normal-weight group costs the least and both underweight and overweight-obesity groups cost more. The median medical costs of the six weight groups were $3184, $2225, $2388, $2801, $3182, and $3753, respectively, with statistical differences existing between any two groups of the last five categories. The underweight groups, especially in females, were not consistent with the guidelines in the two young groups (ages 19 44 and 45-54). An inconsistent relationship between medical costs and BMI groups was seen in the oldest males (age 75+). CONCLUSIONS: The six weight groups defined by the 1998 NHLBI guidelines are consistent with concurrent medical costs. Except for the underweight group (BMI < 18.5 kg/m2), medical costs gradually increased with BMI. Given that the prevalence of obesity continues to increase in western countries, effective weight control programs would help avoid a substantial amount of medical costs associated with overweight/obesity and related diseases. PMID- 12545587 TI - Process evaluation of a monitoring log system for community coalition activities: five-year results and lessons learned. AB - PURPOSE: Coalitions can be a successful way to promote healthy initiatives throughout a community. To properly measure the success of coalition-based interventions, it is important to conduct a process evaluation of coalition activities and establish a system for evaluating outcomes. This article describes a process evaluation of a monitoring and feedback system for community coalitions targeting chronic disease risk reduction. METHODS: Community coalitions in six rural, southeast Missouri counties collaborated with the Missouri Department of Health and Saint Louis University to track coalition events using the process described in the Centers for Disease Control and Prevention manual Evaluating Community Efforts to Prevent Cardiovascular Diseases. SUMMARY: First, we describe the methodology used to monitor monthly activities and to evaluate satisfaction with the process. Next, we discuss the data that resulted from the monitoring system and interviews with recorders. Third, we discuss changes made to the monitoring system and lessons learned along the way. Finally, we end with recommendations for incorporating this monitoring system in community practice. CONCLUSIONS: When used properly, this system is an effective way of evaluating and promoting sustainability of community interventions. PMID- 12545588 TI - ADA compliance and the accessibility of physical activity facilities in western Oregon. AB - OBJECTIVES: One of the mandates of Title III of the Americans With Disabilities Act (ADA) is to address the accessibility of public facilities for all people. The aim of this study was to determine how compliant physical activity facilities in western Oregon were with regard to Title III of the ADA. Comparisons were also made with the findings of a study conducted 5 years prior in a different geographic region. METHODS: On the basis of direct observations and physical measurements taken during on-site visits to 50 physical activity facilities located in western Oregon during 2000-2001, facility compliance with Title III of the ADA was characterized in terms of 10 structural domains. RESULTS: No facility was found to be 100% ADA-compliant. Exterior entrance/doors (90%) and telephone accessibility (88%) were the areas where compliance was highest, whereas accessibility to and around exercise equipment (8%) and customer service desk (37%) were areas lowest in compliance. Relative to a previously published study, the facilities in western Oregon showed greater rates of ADA compliance in six of the 10 structural domains evaluated. CONCLUSIONS: There is an on-going need and legal mandate for increasing the accessibility of physical activity facilities. Furthermore, from a social-ecological perspective, the environmental constraints identified in this study might be limiting factors in efforts aimed at increasing individuals with disabilities' physical activity involvement. PMID- 12545590 TI - Is it real or is it memorex? Discerning whether job-related stress or mental illness is causing physician impairment. PMID- 12545589 TI - Building capacity for heart health promotion: results of a 5-year experience in Nova Scotia, Canada. AB - PURPOSE: To present the outcomes of a capacity-building initiative for heart health promotion. DESIGN: Follow-up study combining quantitative and qualitative methods. SETTING: The Western Health Region of Nova Scotia, Canada. SUBJECTS: Twenty organizations, including provincial and municipal agencies and community groups engaged in health, education, and recreation activities. INTERVENTION: Two strategies were used for this study: partnership development and organizational development. Partnership development included the creation of multilevel partnerships in diverse sectors. Organizational development included the provision of technical support, action research, community activation, and organizational consultation. MEASURES: Quantitative data included number and type of partnerships, learning opportunities, community activation initiatives, and organizational changes. Qualitative data included information on the effectiveness of partnerships, organizational consultation, and organizational changes. RESULTS: Results included the development of 204 intersectoral partnerships, creation of a health promotion clearinghouse, 47 workshops attended by approximately 1400 participants, diverse research products, implementation of 18 community heart health promotion initiatives, and increased organizational capacity for heart health promotion via varied organizational changes, including policy changes, fund reallocations, and enhanced knowledge and practices. CONCLUSIONS: Partnership and organizational development were effective mechanisms for building capacity in heart health promotion. This intervention may have implications for large-scale, community-based, chronic-disease prevention projects. PMID- 12545591 TI - Discovering the cause of syncope. A guide to the focused evaluation. AB - Syncope has many causes. Although the most common ones are generally benign and do not require extensive evaluation, cardiac syncope has high rates of recurrence and death. In this article, Dr Hauer describes how physical examination and electrocardiographic findings can guide further evaluation to provide the largest diagnostic yield at the lowest cost. PMID- 12545592 TI - Penicillin resistance in pneumococcal pneumonia. Antibiotics with low resistance potential are effective and pose less risk. AB - Antibiotic resistance is a potential problem around the world. Among the bacteria that cause community-acquired pneumonia (CAP), resistance in Streptococcus pneumoniae is a primary concern. Resistance can occur through genetic mutations in the bacterial strain itself or can be acquired through use of some antibiotics that have a high resistance potential. In this article, Dr Cunha explores the misperceptions about antibiotic resistance and its occurrence, as well as the most appropriate therapy for CAP in the clinical setting. PMID- 12545594 TI - The challenge of nonresolving pneumonia. Knowing the norms of radiographic resolution is key. AB - Pneumonia that fails to resolve after 10 to 14 days of antibiotic therapy can lead physicians to call for consultation or unnecessary invasive diagnostic procedures. Understanding the infectious and noninfectious causes of pneumonia and their normal times to resolution is enormously helpful in the judicious evaluation of and timely intervention in this very challenging condition. PMID- 12545593 TI - Community-acquired pneumonia in immunocompromised patients. Opportunistic infections to consider in differential diagnosis. AB - Immunocompromised persons are at increased risk for a large group of infections that are either uncommon or much less severe in the immunocompetent host. These opportunistic infections broaden the diagnostic considerations in differential diagnosis of community-acquired pneumonia in patients with immunodeficiencies. This article highlighted epidemiologic factors, clinical presentations, and treatment options for four selected opportunistic infections that represent varied classes of pathogens: nematodes (S stercoralis), mycoses (C neoformans), bacteria (P aeruginosa in patients with HIV infection), and viruses (measles virus). PMID- 12545595 TI - Patient notes: hair loss. PMID- 12545596 TI - Issues in managed care. PMID- 12545597 TI - Medicaid confronts a changing managed care marketplace. AB - After two decades of concerted efforts, more than one-half of all Medicaid beneficiaries are now enrolled in managed care arrangements. Most States appear strongly committed to continued reliance on managed care, but the contemporary managed care marketplace is undergoing a number of significant changes. We describe how several of these developments are being revealed in commercial managed care and discuss implications for Medicaid purchasers and beneficiaries. State Medicaid agencies will have to adapt managed care strategies to respond to the evolving products and practices of managed care plans and their interest in public sector product lines. PMID- 12545599 TI - Managed care and dually eligible beneficiaries: challenges in coordination. AB - This article describes administrative issues and beneficiary perspectives on the delivery of medical services under Medicare+Choice (M+C) and/or Medicaid managed care organizations (MCOs) for dually eligible beneficiaries. We interviewed staff at nine health plans in four market areas in 2000 and 2001, and conducted beneficiary focus groups in 2001. The study reveals beneficiary confusion about the relationship between their dual coverage and managed care enrollment, and problems with care and benefit coordination across these arrangements, based on regulatory and administrative obstacles to effective benefit and care coordination for beneficiaries enrolled in these varied managed care arrangements. PMID- 12545600 TI - Payment policy and competition in the Medicare+Choice program. AB - Over the last 2 years, Medicare+Choice (M+C) plans raised premiums and reduced benefits to an unprecedented degree, arguing that these were unavoidable consequences of inadequate payments. We investigate plan premium and benefit decisions, taking advantage of a natural experiment to separate the influences of payment rates, the intensity of interplan competition, and the underlying cost of providing coverage. We find that the effects of competition are comparable in importance to the effects of payment rates, confirming empirically that it is possible for the Medicare Program to improve benefits without increasing spending or shifting additional costs to beneficiaries. PMID- 12545598 TI - Adjusted clinical groups: predictive accuracy for Medicaid enrollees in three states. AB - Actuarial split-sample method were used to assess predictive accuracy of adjusted clinical groups (ACGs) for Medicaid enrollees in Georgia, Mississippi (lagging in managed care penetration), and California. Accuracy for two non-random groups- high-cost and located in urban poor areas--was assessed. Measures for random groups were derived with and without short-term enrollees to assess the effect of turnover on predictive accuracy. ACGs improved predictive accuracy for high-cost conditions in all States, but did so only for those in Georgia's poorest urban areas. Higher and more unpredictable expenses of short-term enrollees moderated the predictive power of ACGs. This limitation was significant in Mississippi due in part, to that State's very high proportion of short-term enrollees. PMID- 12545601 TI - Impact of Medicare managed care market withdrawal on beneficiaries. AB - The 2001 Survey of Involuntary Disenrollees was conducted to investigate the impact of Medicare+Choice (M+C) plan withdrawals on Medicare beneficiaries. Eighty-four percent of a total of 4,732 beneficiaries whose Medicare managed care (MMC) plan stopped serving them at the end of 2000 responded to the survey. Their responses indicated that the withdrawal of plans from Medicare affected beneficiaries in terms of concerns about getting and paying for care, increased payments for premiums and out-of-pocket costs, and changes in health care arrangements. Of particular concern were the impacts on those in vulnerable subgroups such as the disabled, less educated, and minorities. PMID- 12545602 TI - Medicare health maintenance organization benefits packages and plan performance measures. AB - This article reports the results of an analysis of the relationship between supplemental benefits offered by Medicare+Choice (M+C) plans and their plan performance ratings. We examined two measures of plan performance: (1) plan ratings as reported in the Medicare Managed Care (MMC) Consumer Assessment of Health Care Study (CAHPS), and (2) disenrollment rates. The results of our analysis indicated that variations in plan supplemental offerings have little impact on enrollees' plan performance ratings--both overall ratings and access to care measures. Furthermore, disenrollment rates were found to be more sensitive to the availability of alternative M+C plans, either in general, or for specific benefits than to variations in benefit offerings. PMID- 12545607 TI - Ask your patient to sign this! PMID- 12545606 TI - My Christmas bell. PMID- 12545603 TI - Medicare+Choice individual and group enrollment: 2001 and 2002. PMID- 12545605 TI - Comparison of Medicare risk HMO and FFS enrollees. PMID- 12545608 TI - Why Dr. Kooyer had to move. PMID- 12545604 TI - Rural hospital wages and the area wage index. AB - We examined data on hospital hourly wages and the prospective payment system (PPS) wage index from 1990 to 1997, to determine if incremental changes to the index have improved its precision and equity as a regional cost adjuster. The differential between average rural and urban PPS hourly wages has declined by almost one-fourth over the 8-year study period. Nearly one-half of the decrease is attributable to regulatory and reporting changes in the annual hospital wage survey. Patterns of within-market wage variation across rural-urban continuum codes identify three separate sub-markets within the State-level aggregates defining rural labor markets. Geographic reclassification decisions appear to eliminate one of the three. Remaining systematic within-market rural wage differences work to the reimbursement advantage of hospitals in the smaller and more isolated communities. PMID- 12545609 TI - We nailed an HMO for $6 million. PMID- 12545610 TI - If one patient threatens another. PMID- 12545611 TI - Under siege. The proposed sale of Health Midwest has landed the not-for-profit in the fight of its life. At stake--control of an $800 million foundation. AB - Two state attorneys general, one goal. Kansas' and Missouri's top lawyers are teaming up against the board of directors of Health Midwest, which is battling to survive and control an estimated $800 million foundation that would result from the sale of its assets to HCA. Kansas Attorney General Carla Stovall (left) and her Missouri counterpart have asked the courts to dissolve Health Midwest and oust its board. PMID- 12545612 TI - Healthcare leaders plan way to help patients pick hospitals. New voluntary reporting system calls for facilities to disclose information about effectiveness of care. PMID- 12545613 TI - More changes. Disclosure heralds new era for keeping medical records. PMID- 12545614 TI - Grim outlook. Increased Medicaid spending may force states to cut costs, scale back benefits. PMID- 12545615 TI - Leaky umbrella. Invitees not willing to pay for inclusion in AMA. PMID- 12545616 TI - More bad news. Tenet faces JCAHO surprise visits, management critic. PMID- 12545617 TI - High-tech treatment. Federal grant allows HIV/AIDS patients to receive treatment through telemedicine. PMID- 12545618 TI - AHA's specialty hospital task force suggests policy changes. Physician investors consider proposals unfair control of competition. PMID- 12545619 TI - The erector set. Many hospitals are spending big to rebuild and save in the long run. PMID- 12545620 TI - A strong attraction. As hospitals seek 'magnet' status to retain nurses and improve care, some see the program as little more than a pricey marketing gimmick. PMID- 12545621 TI - Telltale signs your board of directors may be providing ineffective governance. PMID- 12545622 TI - [Salvage radical prostatectomy after radiotherapy failure in localized prostatic cancer]. AB - OBJECTIVE: The objective of this study was to evaluate salvage radical prostatectomy after failure of radiotherapy for localized prostate cancer. MATERIAL AND METHOD: The literature published between 1984 and 2002 was reviewed by a MEDLINE search using the following key words: prostate cancer, radiotherapy, salvage radical prostatectomy. RESULTS: A total of 404 cases of salvage radical prostatectomy were identified, including 287 radical prostatectomies and 117 radical cystoprostatectomies. Patient selection criteria for this type of surgery were: good general state with a life expectancy of at least 10 years, tumour recurrence with no signs of local or distant extension, histological evidence a minimum of 12 months after radiotherapy and PSA level less than 10 ng/ml before radiotherapy and surgery. The mean rectal injury rate was 7% (range: 0% to 28%) and the mean incontinence rate was 37% (range: 0% to 80%). Histologically, the mean rate of prostate-confined tumours was 34% (range: 20% to 60%), a mean seminal vesicle invasion rate of 36% (range: 25% to 50%) and a mean positive surgical margins rate of 36% (range: 15% to 67%). This second-line treatment was effective in terms of long-term survival, with a specific 10-year survival of 70%. CONCLUSION: Radical prostatectomy after failure of radiotherapy is associated with considerable morbidity, but can achieve a good 10-year survival rate in carefully selected patients. PMID- 12545623 TI - [Hyperbaric oxygen therapy and radiation-induced hemorrhagic cystitis]. AB - Radiotherapy alone or in combination with other modalities is used in the treatment of a large number of pelvic tumours of urological or gynaecological origin. Despite constant progress in this field, medium-term and long-term complications remain frequent and often require difficult long-term management. Radiation cystitis is one of the most frequent complications and directly concerns urologists. Among the various treatment options for haemorrhagic cystitis, hyperbaric oxygen therapy appears to give good short-term and medium term results. It is currently reserved for cases refractory to the standard treatments for radiation cystitis. PMID- 12545624 TI - [Treatment of renal tumors with radiofrequency: preliminary results]. AB - OBJECTIVE: To study the feasibility and the innocuity of the treatment by radiofrequency of small renal tumours. MATERIAL AND METHODS: From June 2000 to September 2001, 10 renal tumours in 6 patients were treated by radiofrequency. The mean age of the patients was 57 years. The mean tumour diameter was 20 mm. Indications for partial surgery were 3 solitary kidneys, one patient with chronic renal failure, one patient with Von Hippel Lindau disease and one patient in whom several angiomyolipomas were discovered in the right kidney. 4 patients were treated via a lumbar incision and 2 were treated percutaneously (one with ultrasound guidance and the other with computed tomography guidance). Patients were reviewed by imaging (CT and/or MRI) with injection of contrast agent to demonstrate complete devascularization of the tumour. RESULTS: Treatment lasted an average of 180 minutes. The intraoperative complication rate was 16%. The mean hospital stay was 9 days (4 days for the percutaneous route). Nine tumours (5 patients) were devascularized after a single session. One patient (treated via the percutaneous route under ultrasound guidance) showed persistence of tumour tissue (identified by CT). A second session, performed 4 months later, allowed complete devascularization of the tumour. The mean follow-up is 13 months. CONCLUSION: This study demonstrates the feasibility and safety in terms of renal function of radiofrequency treatment of small renal tumours. However further studies are necessary to precisely define the radiological criteria of tumour necrosis and to evaluate the long-term results of this method (outcome of necrotic tumour tissue, metastatic risk, cancer recurrence along the needle track in the case of a percutaneous approach). This treatment must be reserved for selected patients. PMID- 12545625 TI - [Lumboscopic treatment of simple renal cysts: initial experience with 17 cases]. AB - OBJECTIVE: To evaluate the feasibility of lumboscopic treatment of simple renal cysts. MATERIAL AND METHODS: From January 1999 to January 2002, 17 patients with a mean age of 40 years were operated for symptomatic renal cysts via a retroperitoneal approach. All patients were evaluated by preoperative ultrasonography and CT. A Bosniak type I cyst was diagnosed in 10 cases and a Bosniak type II cyst was diagnosed in 7 cases. The mean cyst diameter was 7 cm (range: 5 to 15 cm). RESULTS: All patients were operated via an exclusively retroperitoneal approach. The mean operating time was 50 min (range: 40 mm-120 mm). The estimated mean blood loss was 80 cc. No conversion was necessary and the mean length of hospital stay was 1.5 days. All cysts were histologically benign. All 17 patients are asymptomatic (mean follow-up = 11 months) with no radiological signs of recurrence. CONCLUSION: Lumboscopic treatment of simple renal cysts appears to be a feasible, safe, effective and satisfactory technique in terms of morbidity, operating time, complications and blood loss. PMID- 12545626 TI - [Impact of the use of double J stents in renal transplantation on incidence of urologic complications and urinary infection]. AB - OBJECTIVE: The authors evaluated the impact of systematic placement of a double J stent on urological complications and postoperative urinary tract infection. MATERIAL AND METHODS: 188 renal transplantations were performed between January 1991 and December 2001. All patients underwent Lich-Gregoire ureterovesical anastomosis. A double J stent was not used in one group (G1) of 110 patients and a double J stent was systematically inserted in another group (G2) of 78 patients and was removed after 3 weeks. Urine culture was performed in all patients. RESULTS: 11 patients of group 1 developed urinary fistula and one patient developed obstruction of the ureterovesical anastomosis. No urological complication was detected in group 2. The postoperative urinary tract infection rate was 47.2% in group 1 versus 48.7% in group 2. CONCLUSION: Systematic double J stenting appears to decrease the incidence of urological complications after renal transplantation without increasing the incidence of postoperative urinary tract infection. PMID- 12545627 TI - [Ureterocele in adults complicated with calculi: diagnostic and therapeutic features. Report of 20 cases]. AB - STUDY OBJECTIVE: The authors study the various diagnostic and therapeutic aspects of adult ureterocele and evaluate the place of endoscopic treatment. MATERIALS AND METHODS: This retrospective study was based on 20 cases of ureterocele in adults complicated by stones and observed between January 1990 and December 2001. This series consisted of 8 men and 12 women with a mean age of 48.3 years (range: 24-75 years). The clinical features were dominated by low back pain. The diagnosis of ureterocele was based on intravenous urography in 19 patients and was discovered on endoscopy in one patient. The ureterocele affected a single ureter in 16 cases (80%) and a duplex ureter at the expense of the upper renal segment in 4 cases. All patients were treated by endoscopic horizontal meatotomy with stone fragmentation and extraction. The mean operating time was 27 min (range: 18-58 min). RESULTS: The postoperative course was uneventful in 19 patients. One patient developed postoperative sepsis. All patients were reviewed clinically and radiologically at 3 months and at 6 months. Satisfactory results were obtained in every case with no residual stones and resolution of low back pain. Only one patient presented vesicoureteric reflux which had resolved on the follow-up urethrocystography performed at 6 months. CONCLUSION: Adult ureterocele complicated by stones is a well tolerated, rare entity that can often be easily diagnosed. Endoscopic meatotomy is easy to perform and gives good results. The associated stones constitute an additional argument in favour of endoscopic treatment. PMID- 12545628 TI - [Prognostic value of tumor location of urothelial tumors of the bladder, after total cystectomy]. AB - OBJECTIVE: Comparison of the various treatment protocols for bladder cancer requires identification of survival prognostic factors. Some clinical parameters, such as tumour sub-site, have still not been studied. The authors therefore analysed the prognostic value of this factor, in combination with known prognostic factors, after radical cystectomy for urothelial bladder tumour. MATERIALS AND METHOD: 120 patients were treated for pure urothelial bladder tumour with pelvic lymphadenectomy, between January 1980 and January 1999, in a single centre. The prognostic value of the various sub-sites was studied by univariate and multivariate survival analysis. RESULTS: The 5-year survival decreased from 62% to 19% in the case of a lesion involving the bladder dome (p = 0.0001). Multivariate analysis demonstrated the following independent prognostic factors: local invasion pT < or = 3 (p = 0.003), lymph node involvement (p = 0.05) and involvement of the dome (p = 0.002). Other tumour sub-sites had an effect on survival on univariate analysis that was not confirmed on multivariate analysis. CONCLUSION: Although stages pT and pN are prognostic factors recognized by all studies, invasion of the bladder dome appears to have a major independent influence on survival. This result must be confirmed by other studies, in view of its potential influence for determination of homogeneous patient groups in controlled trials. PMID- 12545630 TI - [Exclusively extraperitoneal laparoscopic radical prostatectomy]. AB - OBJECTIVE: To evaluate the technical feasibility, efficacy, and intraoperative and postoperative morbidity of exclusively extraperitoneal laparoscopic radical prostatectomy (RP). MATERIAL AND METHODS: From August 2000 to December 2001, 35 patients have had an extraperitoneal laparoscopic radical prostatectomy. The operation comprised an exclusively extraperitoneal 6 cm mini-laparotomy with the use of a camera and laparoscopic instruments. RP is performed via an anterograde approach, starting with release of the bladder neck. 35 patients (pts) were operated from 1st August 2000 to 1st December 2001. RESULTS: RP was performed via this approach in every case without the need for conversion or enlargement of the incision. The mean operating time was 135 min. No intraoperative complications were observed. Mean blood loss was 210 cc (< 100-350 ml). No patient required transfusion. Discharge was possible on the 5th day in 22 pts the 6th day in 10 pts and the 7th day in 3 pts. Oral feeding was resumed on D1. The bladder catheter was left in place for an average of 4.5 days. Oncological Results: 5 pts presented capsular effraction (PT3--negative fixed margin), 1 pt presented a positive margin at the apex (perineural infiltration). Follow-up is more than three months for all patients. PSA is < 0.1 in 34 of the 35 pts. Continence: at one month, 29 out of 35 pts used 0 to 1 protections/day, at the 3rd month, 33/35 no longer used any protection. Only 2 out of 35 pts used protection beyond 4 months during intense physical efforts. CONCLUSION: Radical prostatectomy can be performed satisfactorily by this technique which combines the advantages of the two surgical procedures (laparoscopic and open). The operating time is slightly longer than for conventional surgery. The technique appears to be perfectly reproducible, with a rapid learning curve. Atraumatic dissection of the prostatic apex appears to give satisfactory results in terms of urinary continence and postoperative pain is minimal allowing rapid mobilization of the patient. PMID- 12545629 TI - [Peri-prostatic nerve block at the level of the apex versus intrarectal lidocaine gel]. AB - INTRODUCTION: After a review of the contradictory results of studies concerning periprostatic nerve block, the authors decided to prospectively evaluate the efficacy of periprostatic nerve block in the apex compared to the use of endorectal lidocaine gel. METHODS: From January 2001 to January 2002, 110 patients underwent prostatic biopsies. Patients were randomized to receive 10 cc of 2% lidocaine gel in the endorectal (group 1), or 10 cc of 1% lidocaine solution injected into the apex to induce bilateral periprostatic nerve block (group 2). Pain was evaluated by a 10-point linear visual analogue pain scale (VAS), and a 5-point digital visual scale (DVS). RESULTS: 96 patients were included in the study: 43 in group 1 and 53 in group 2. The mean pain score on the VAS was 2.76 +/- 1.69 and 1.73 +/- 1.26 for groups 1 and 2, respectively (p = 0.001). The DVS score was 2.26 +/- 0.82 and 1.62 +/- 0.56 for groups 1 and 2, respectively (p < 0.001). No significant difference was observed for mean age (p = 0.348), prostatic volume (p = 0.899), PSA level (p = 0.932) and complications between the two groups. The number of biopsies per patient was significantly higher in group 2 (p = 0.006), with significantly lower pain scores on each of the 2 scales. CONCLUSIONS: Apical periprostatic nerve block ensured better pain control during transrectal prostate biopsy, compared to the use of endorectal lidocaine gel, with no increase in the complication rate. PMID- 12545631 TI - [Ejaculatory profile: a pressure of 5 meters H2O at the level of the bladder neck during ejaculation]. AB - INTRODUCTION: The existence of a pressure gradient in order to prevent retrograde ejaculation in men with no ejaculatory disorders has always been postulated, but without any scientific evidence. MATERIAL AND METHOD: The profile of the prostatic urethra was recorded during ejaculation in 5 men with no ejaculatory disorders using a 10 F balloon catheter with 16 pressure channels, situated in pairs every 5 mm, starting just below the balloon in the bladder neck and extending as far as the external urethral sphincter. RESULTS: The pressure in the proximal part of the proximal urethra was 500 cm H2O in the five men, but this pressure did not exceed 400 cm H2O distally as far as the colliculus seminalis. CONCLUSION: The authors present a new method for recording the urethral pressure profile during ejaculation (ejaculatory profile). This study provides a better understanding of the mechanisms of normal ejaculation and could be useful for the evaluation of disorders of ejaculation. PMID- 12545632 TI - [TVT: prospective study comparing pre- and postoperative clinical, urodynamic and quality of life criteria]. AB - INTRODUCTION: This prospective study on TVT compared preoperative and postoperative urodynamic in order to evaluate the effects of TVT on the detrusor sphincter synergy, and quality of life criteria by means of a validated questionnaire. MATERIAL AND METHOD: From August 2000 to March 2002, the authors conducted a prospective study on two sites, Nice and Marseille, on 37 patients. The inclusion criteria were clinical and urodynamic. A questionnaire evaluating the severity of incontinence and its impact on daily life was completed preoperatively and postoperatively. Thirty TVTs were placed under local anaesthesia and seven TVTs were placed under general anaesthesia. Patients were reviewed at least 6 months (mean: 9 months; range: 6 to 19 months) after the operation. Statistical analysis was based on a test for paired series, patient by patient, with an a risk of 5%. RESULTS: Eight cases of unilateral bladder perforation (21.6%) were treated by catheterization for 48 hours and 6 cases of transient acute urinary retention (16.2%) were also treated by catheterization. No haemorrhagic or infectious complications were observed. Thirty one (83.7%) patients were completely cured of their incontinence, 5 (13.5%) were improved and there was one failure. Six (75%) of the 8 patients with a urethral closing pressure less than 30 cm H2O, 6 were cured and 2 (25%) obtained a marked improvement. All criteria were compared in the same patient preoperatively and postoperatively. The maximum flow rate on uroflowmetry was significantly lower postoperatively, but no difference was observed for preoperative and postoperative detrusor sensitivity, urodynamic bladder capacity or urethral closing pressure. No de novo detrusor instability was observed. Mean severity scores and impact of incontinence scores were much lower postoperatively. CONCLUSION: The efficacy of TVT does not appear to be correlated with the severity of sphincter incompetence and did not modify the detrusor-sphincter synergy of the patients of our study. The use of a questionnaire appears to be useful and should be recommended in order to more accurately evaluate the efficacy of the surgical procedure and the improvement of the patient's quality of life. PMID- 12545633 TI - [Efficacy of suburethral TVT in cases of stress urinary incontinence associated with severe sphincter insufficiency. Report of 22 cases]. AB - INTRODUCTION: The TVT system has revolutionized the management of female stress urinary incontinence (SUI). Less favourable results have been observed in patients with urinary incontinence associated with a low maximum urethral closing pressure (MUCP). The objective of this prospective study was to evaluate the efficacy of TVT in this situation. MATERIAL AND METHODS: From October 1999 to June 2001, 22 patients between the ages of 53 and 84 years (median: 66.7 years) were operated for SUI with an MUCP < 25 cm H2O (mean: 17.6 cm H2O, range: 6 to 22 cm H2O) present for more than 8 months (range: 8 to 48 months). Three patients also had associated urgency. The Ulmstem manoeuvre was positive in every case and the Bonney manoeuvre was positive in 19 patients. Insertion of TVT was associated with sacral colpopexy in two patients. Success of treatment was defined by complete absence of incontinence and voiding disorders after the operation. RESULTS: One patient was lost to follow-up. The median follow-up was 7.5 months (range: 3 to 27 months). The mean duration of bladder catheterization was 1.3 days (range: 1 to 5 days). Fifteen patients were completely cured by TVT (71.4%). In these cases, uroflowmetry and post-voiding volume were not affected by the operation. Treatment was ineffective in 6 patients (4 cases of uncontrolled leaks, one case of de novo urge incontinence, one case of postoperative dysuria with a post-voiding residue of 120 ml). CONCLUSION: The TVT system appears to be an effective method for the treatment of most patients with SUI associated with severe manometric sphincter incompetence. However, these favourable results need to be confirmed by a longer follow-up. PMID- 12545634 TI - [Survey on satisfaction of patients with Malone continent cecostomy]. AB - OBJECTIVE: A questionnaire to assess their level of satisfaction was sent to 28 patients who underwent a continent cecostomy for antegrade enema (Malone procedure). MATERIAL AND METHOD: All the patients presented with neurogenic bladder and a severe constipation due to spina bifida in 21 cases, anorectal malformation in 4, cloacal malformation in 1, sacral agenesis in 1 and post myelitis paraplegia in 1. The mean age at surgery was 15 years. The mean follow up is 3 years and 8 months. Surgery on the bladder was done in 18 patients of which 10 at the same stage. Urinary continence between bladder catheterizations occurred in 5 patients with isolated cecostomy. RESULTS: 24 patients answered the questionnaire. Among the remaining 4, 1 has a too short follow-up, 1 refused the enemas and his cecostomy definitely closed and 2, when seen at clinics, have an excellent result. Enemas are done twice a week, they last on average one hour with an average of 3 litres of liquid, usually tap water. All the 24 patients consider to have benefited from the operation mainly regarding their personal, family and social wellbeing. All have acquired anal continence, only 3 have some soiling between enemas. The main reported drawbacks concern pain at passing of stools (14 cases) well improved by addition of Colopeg in the enema, fatigue (21 cases) or headache (5 cases) after enemas. These troubles, probably due to a temporary hyponatremia, are well improved by addition of table salt in the water. CONCLUSION: This enquiry established the high degree of satisfaction given by the Malone procedure in the treatment of severe constipation frequently associated with a congenital neurogenic bladder. Despite some drawbacks this solution must be considered in such situation. PMID- 12545635 TI - [Role of fetal urine sampling in the prenatal diagnosis of malformative uropathies]. AB - OBJECTIVE: To define the diagnostic and therapeutic value of foetal urine sampling (FUS). MATERIAL AND METHOD: Between 1996 and 2001, FUS was performed in 16 patients for three indications: assessment of renal function (Group 1, n = 12), diagnosis of a cystic mass (Group 2, n = 2), prevention of dystocia (Group 3, n = 2). The anomaly was detected by ultrasound. Urine was aspirated from the two renal pelves and/or bladder, or the cystic mass in the other cases. b2 microglobulin and urinary sodium were assayed and karyotyping was performed at the same time. RESULTS: 20 FUS were performed In 4 cases, two samples were taken at an interval of 2 weeks. No complications were observed. Group 1: 9 pregnancies were terminated for potential renal failure (6 posterior urethral valves (PUV), 1 Prune Belly syndrome, 2 cases of recessive polycystic kidney disease (PKD). Three pregnancies were continued for 2 foetuses with normal renal function (1 PUV, 1 ureterocele) and one foetus died with severe renal failure (bilateral hydronephrosis). Group 2: one termination of pregnancy was performed in a case of hepatic cyst with portobiliary dysplasia and a giant bladder diverticulum was operated. Group 3: in both cases, evacuating aspiration allowed normal delivery of one live infant (left polycystic dysplasia), and one neonatal death from megabladder-megacolon syndrome. DISCUSSION: The value of FUS is controversial. We usually performed this examination for assessment of renal function. The results of FUS are useful for management decisions when they are in favour of potential impaired renal function. They usually confirmed the ultrasound results that already indicated a poor prognosis and they did not change the decision already based on ultrasound findings in any of our cases. For the diagnosis of cystic masses, after drainage of the mass, FUS facilitated investigation of the urinary tract and adjacent organs. When FUS was performed during labour, it reduced the size of the mass and the abdomen, allowing vaginal delivery with a better ventilatory adaptation of the infant. PMID- 12545636 TI - [Treatment of phimosis with locally applied 0.05% clobetasol propionate. Prospective study with 108 children]. AB - OBJECTIVE: This study was designed to evaluate the efficacy of topical application of corticosteroid cream in the treatment of phimosis in children and to compare the results obtained with those reported in the literature. MATERIALS AND METHODS: In the context of a prospective study conducted between January 1997 and November 2001, 108 children with a mean age of 5.9 years, presenting with true phimosis preventing retraction of the foreskin, were treated by twice-daily application of 0.05% clobetasol propionate cream for 10 days. The results were evaluated at one month and then every six months by a urologist in the outpatients department. A second course of treatment identical to the first was proposed in the case of failure or partial efficacy. RESULTS: 96 children were evaluated with a mean follow-up of 14 months (range: 6 to 24 months). Seventy nine children (82%) are cured after a first course of treatment. In the remaining 17 children, 2 were treated by circumcision and 15 received a second course of treatment resulting in cure in another 9 cases. The final cure rate was therefore 92%. Among the six failures, 4 children were circumcised, corresponding to a total of 6 circumcisions out of this series of 96 cases of phimosis (6%). No local or systemic adverse effects related to steroid therapy were observed and no recurrence of phimosis was observed during the study. CONCLUSION: This study confirms the efficacy and safety of local steroid therapy in the treatment of phimosis in children. Surgical treatment of phimosis should therefore be reserved for failures of medical treatment and has now become much less frequent. PMID- 12545637 TI - [Breast metastasis of kidney cancer. Diagnostic and therapeutic features]. AB - The breast is a rare site of metastasis from renal cancer and the preoperative diagnosis is often difficult. The treatment recommended for this type of metastasis is tumourectomy associated with frozen section examination, while mastectomy and lymph node dissection are unnecessary. The prognosis is often poor with a life expectancy rarely exceeding one year. The authors report a case of isolated right breast metastasis occurring two and a half years after left radical nephrectomy for renal cell carcinoma. Tumourectomy and frozen section examination were performed. The patient died four years later in a context of disseminated metastases despite immunotherapy. PMID- 12545638 TI - [Hydatid cyst on horseshoe kidney]. AB - Hydatid cyst on horseshoe kidney is a rare entity in temperate climates and the diagnosis is often delayed. The clinical features are marked by signs of compression of adjacent organs, mass syndrome and pain. The diagnosis is based on ultrasonography and CT. Arteriography can be useful to study the blood supply of the horseshoe kidney. Treatment is surgical and must remain conservative, as this is a benign disease. In a small percentage of cases, the hydatid cyst can be mistaken for a cystic malignant renal tumour, but the diagnosis is confirmed by serology. The course is usually favourable. PMID- 12545639 TI - [Bilateral primary adrenal lymphoma]. AB - The adrenal gland is a rare site of non-Hodgkin's lymphoma, as only about 70 cases have been reported in the literature, usually with bilateral involvement. Most tumours have a high grade histology, almost always with the B phenotype. Medical imaging is nonspecific and biopsy remains the most reliable diagnostic method. Chemotherapy is the treatment of choice, but the prognosis remains poor in the majority of cases, although long-term survivals have been described. The authors report a case of bilateral high-grade lymphoma of the adrenal glands in a 31-year-old patient presenting with acute adrenal insufficiency. Imaging demonstrated large bilateral adrenal masses, and surgical biopsy of the adrenal gland and staging confirmed the diagnosis of bilateral primary adrenal lymphoma. After corticosteroid replacement therapy, treatment consisted of primary CHOP chemotherapy administered for 9 cycles, followed by external beam radiotherapy delivered at a dose of 40 Gy. After 3 months of follow-up, the patient was still alive, with partial response, with an overall survival of 15 months. PMID- 12545640 TI - [Pseudotumor form of urinary tuberculosis]. AB - Urogenital tuberculosis is an increasingly frequent serious disease. The diagnosis is often delayed due to the marked clinical polymorphism, leading to serious sequelae. The diagnosis of typical forms is easy, but some forms are misleading and can lead to an incorrect diagnosis. The authors report a pseudoneoplastic form of urogenital tuberculosis in a young patient that was initially diagnosed as lymphoma. In the light of this case and a review of the literature, the authors emphasize the diagnostic difficulties of these forms and the treatment modalities. PMID- 12545641 TI - [Psoas tumors]. AB - The authors report a case of benign tumor of the psoas muscle occurring in a 36 year-old man. The diagnosis was based on computed tomography and the patient underwent tumorectomy by an extrapoeritoneal approach. The authors emphasize the ratity f psoas tumors that are usually malignant and the advantage of computed tomography and percutaneous biopsy. Treatment must be incorporated into a multidisciplinary approach in which surgery plays a leading role and must be based on the treatment of soft tissue tumors. PMID- 12545642 TI - [Retro-iliac ureter associated with ipsilateral megaureter]. AB - Retro-iliac ureter is a rare congenital abnormality. The authors report a case of retro-iliac ureter associated with ipsilateral megaureter in a 38-year-old patient treated by section of the ureter at the ureterovesical junction, and dissection as far as the iliac pedicle and anterior repositioning of this pedicle, followed by ureterovesical reimplantation. Based on a review of the literature, this appears to be the first published case of this pathological association. PMID- 12545643 TI - [Multifocal urothelial tumors and renal autotransplantation]. AB - The authors report the case of a patient with multiple urothelial tumours only sparing the superior left ureter and the urethra. The complex treatment consisted of a combination of resection of the entire urinary tract, except for the left kidney and its proximal ureter that was transplanted into the right iliac fossa, after ex vivo partial pyelectomy. This complex treatment regimen avoided the need for dialysis with a good functional and oncological result, three years later. PMID- 12545644 TI - [Non-typhus Salmonella urinary tract infections]. AB - Non-typhus Salmonella urinary tract infections are rare. Based on a clinical case and a review of the recent literature, the authors analyse the diagnostic setting and discuss the necessary investigations and treatment modalities. Non-typhus Salmonella urinary tract infections generally occur in a predisposed clinical setting, such as immunodepression or acquired (stones, schistosomiasis) or congenital uropathy. As in the present case, it can also be the first clinical expression of AIDS, which must be systematically investigated. Antibiotic therapy must be continued for at least two weeks and consists of a third generation cephalosporin or fluoroquinolones, but the prognosis is directly related to the underlying clinical condition. PMID- 12545645 TI - [Pelvic pain caused by bladder leiomyoma: diagnostic and radiologic difficulties]. AB - Uterine leiomyoma is a frequent benign lesion. It is usually asymptomatic and can be responsible for acquired dysmenorrhoea in some women, leading to gynaecological assessment. However, the development of this type of muscle lesions arising from the bladder wall and accompanied exclusively by dysmenorrhoea is rare. It is very difficult to study the anatomical relations of the lesion and distinguish it from other pelvic, genital or urinary tract tumours. The authors discuss the diagnostic aspects and therapeutic management in the light of a case of bladder leiomyoma in a 39-year-old woman. PMID- 12545646 TI - [Pseudo-tumor form of bladder scleroderma: unusual cause of hematuria]. AB - The bladder is a rare site of scleroderma. The frequency of bladder involvement is difficult to estimate, as only about thirty cases have been reported in the literature. Scleroderma bladder involvement usually presents in the form of frequency and burning, but may sometimes cause urgency or recurrent urinary tract infections. The authors report a case of bladder scleroderma responsible for severe haematuria in a 56-year-old woman and discuss this rare cause of macroscopic haematuria in the light of the histological findings and a review of the literature. PMID- 12545647 TI - [The future of biomaterials in urology]. AB - The fields of application of implanted biomaterials will continue to grow, ranging from the use of catheters and slings for the genitourinary tract, to the production of tissues and organs. The scale of research has changed over recent years, passing from a macroscopic view to a microscopic view, resulting in a change from mechanical devices, such as prostheses, catheters and slings to the cellular unit and more particularly in vitro cell cultures. New techniques, such as gene therapy and cell reconstruction have been recently introduced into the field of urology. Prospective biocompatibility studies must be conducted to establish appropriate biocompatibility standards adapted to new materials. The development of these new materials required a multidisciplinary approach involving engineers, biologists and physicists. It must be conducted in specialized centres skilled in biocompatibility research and in close collaboration with a basic bioscience and clinical practice. In the future, synthetic biomaterials will have an almost unlimited range of mechanical properties allowing much more specific applications. Another improvement of materials will concern a reduction of the risk of infection, erosion, mineral deposits, migration of particles, secondary reactions, and finally improved durability. However, the long-term properties of synthetic materials will probably not be as good as those of natural or almost natural biomaterials derived from biotechnology by tissue engineering. PMID- 12545648 TI - [Contribution of the latest generation CT in preoperative assessment of kidney cancer]. AB - Technological progress in medical imaging runs in parallel to progress in computers, with an ever-increasing image acquisition rate and increasingly fine image reconstruction. Following the appearance, several years ago, of spiral acquisition CT, the multiplication of detectors (multi-array CT) has opened up new possibilities for investigation, especially in the field of renal tumours, where CT is now the first-line investigation due to the information obtained for characterization, vascular anatomical relations, and staging. The development of partial surgery (in which the detection of satellite lesions is essential) and minimally invasive laparoscopic surgery will be able to take full advantage of this progress in imaging by anticipating operative difficulties (number of tumours, situation and branches of arteries and veins or their collaterals, venous thrombus, relations with the urinary tract). Volume acquisition allows representation of these elements in the various dimensions, facilitating interpretation of the images. PMID- 12545649 TI - [Is bladder colposuspension still indicated in female stress urinary incontinence?]. AB - Female stress urinary incontinence may need to be treated surgically following failure of non-surgical treatments. Many techniques are now available and are selected according to the pathophysiology of incontinence. Vesical colposuspension is considered to be the reference operation for the surgical treatment of female stress urinary incontinence (SUI). New suburethral sling techniques have been available for several years, such as TVT, SPARC, IVS or, even more recently, URATAPE. These new techniques not only allow restoration of continence, but are also minimally invasive and easily reproducible. They also facilitate re-operation in the case of failure or recurrence of incontinence. Currently, although the follow-up of these techniques is only 5 years, they ensure better management of the various types of urinary incontinence (stress, mixed and sphincter incompetence) than vesical colposuspension. PMID- 12545650 TI - [Hematuria caused by nutcracker syndrome: peroperative confirmation of its presence]. AB - Nutcracker syndrome should be considered in the case of left ureteric haematuria based on computed tomography with vascular reconstruction of the hilar region of the kidney. The best confirmation is obtained by studying the pressure gradient between the left renal vein and the inferior vena cava during cavography. Various modalities of surgical treatment have been proposed by a few authors, but have been criticised by some authors who question the clinical reality of this syndrome and the efficacy of treatment. The decision to operate may be difficult and other investigations may be useful to confirm the diagnosis. We report the case of a patient in whom the diagnosis of nutcracker syndrome was confirmed intraoperatively by the immediate appearance of massive, reversible haematuria induced by clamping of the renal vein during vein dissection and augmentation plasty. In the light of this original case, we believe that a preoperative percutaneous haematuria provocation test by temporary obstruction of the left renal vein during venography could allow a more formal diagnosis of nutcracker syndrome when this syndrome is highly suspected. PMID- 12545651 TI - [Repair of parastomal hernia with marlex or prolene prosthesis in patients with Bricker-type urinary diversion]. AB - Creation of a cutaneous stoma can be associated with a number of complications, the most frequent being stenosis and parastomal hernia. A hernia can appear anywhere around the stoma, but it is more frequent in the lateral quadrants; it complicates about 5% of cutaneous stomas. Small hernias are usually well tolerated, but large hernias can be severely disabling when standing and on effort, and can prevent correct positioning of external urine collection appliances. The authors propose a repair technique for larger parastomal hernias via a semicircular incision around the stoma and interposition of an allogeneic mesh prosthesis. PMID- 12545652 TI - [Trans-urethral micro-wave thermotherapy of prostatic benign hypertrophy]. AB - Transurethral microwave thermotherapy is designed to destroy non-malignant tissue by exposure to a cytotoxic temperature ranging from 45 degrees C to more than 70 degrees C. The combination of microwave-induced heat administered transurethrally and protection of the urethra by a cooling circuit allows delivery of sufficient power to induce coagulation of the periurethral prostatic tissues without any form of anaesthesia part from instillation of anaesthetic jelly into the urethra. Treatment takes only 30 minutes. Strict security limits (urethral temperature and rectal temperature) ensure a low complication rate. The therapeutic efficacy of thermotherapy is situated between that of drugs and endoscopic surgery. The effect is lasting. The development of a tailor-made treatment for each patient should increase the response rate of this technique. PMID- 12545653 TI - [Galen (121-201) and the first experimental test of ureteral function]. AB - Galen and Hippocrates are the two leading representatives of Ancient Greek and Western medicine. Galenic medicine retained an indisputable authority for fifteen centuries. Urology occupies a considerable place in Galen's work. Galen was the first to provide experimental proof of ureteric function. PMID- 12545654 TI - [Re: "Treatment of genito-urinary prolapse with spinofixation with polypropylene prosthesis," by P. Montete, F. Gibon, Borgogno C., Coloby P. Prog. Urol., (2002), 12, 517-521]. PMID- 12545655 TI - Proceedings of the First International Meeting on EBM in Musculoskeletal Disorders (e_Musk1). Ferrara, Italy, April 2001. PMID- 12545656 TI - EBM in musculoskeletal diseases: where are we? PMID- 12545657 TI - Research methodology in rheumatology. PMID- 12545658 TI - The Cochrane Musculoskeletal Injuries Group. PMID- 12545659 TI - European guidelines for the management of low back pain. PMID- 12545660 TI - Promoting evidence based orthopaedic surgery. An English experience. PMID- 12545661 TI - The German Arthroplasty Register. PMID- 12545662 TI - Evidence-based lumbar spine surgery. The role of national registration. PMID- 12545663 TI - Registration of hip prostheses at the Rizzoli Institute. 11 years' experience. PMID- 12545664 TI - Managing fractures of the distal radius in adults. Clinical and research implications from systematic reviews of existing trials. PMID- 12545665 TI - Lost to follow-up in a hip prosthesis register. Experience of R.I.P.O. PMID- 12545666 TI - Pain in low-back pain. Problems in measuring outcomes in musculoskeletal disorders. PMID- 12545667 TI - The RoentgenCoach-Rheumatology--a novel tool to enhance efficacy of radiological scoring of rheumatoid arthritis. Results of experimental scoring of 72 cases. PMID- 12545668 TI - Atypicality. One of the intellectual challenges of geriatric medicine. PMID- 12545669 TI - Sores on the index finger. Identifying a patient's pre-existing medical conditions aids in diagnosis. PMID- 12545670 TI - Silent MI. Prevalence and prognosis in older patients diagnosed by routine electrocardiograms. AB - Older men and women with coronary artery disease and silent myocardial ischemia detected by 24-hour ambulatory electrocardiograms (ECGs) have a higher incidence of new coronary events than those with no silent ischemia. The prevalence of clinically unrecognized MI detected by routine ECGs in older persons varies from 21 to 68%. Atypical symptoms associated with acute MI in older persons include dyspnea and neurologic and GI symptoms, as well as chest pain. Older persons with clinically unrecognized MI have a similar or higher incidence of new coronary events and mortality compared with those with recognized MI, which suggests that an aggressive diagnostic and therapeutic approach may be beneficial in these patients. PMID- 12545671 TI - Cardiovascular disease and the aging U.S. population. PMID- 12545673 TI - Act in time to heart attack signs [symbol: see text] action plan. PMID- 12545672 TI - Hypertension. Contemporary challenges in geriatric care. AB - Hypertension has been unequivocally linked to morbid complications such as heart attack, congestive heart failure, renal failure, and stroke. Despite the availability of myriad effective antihypertensive agents, blood pressure remains either untreated or inadequately controlled to even conservative goals in many patients. Only 68.4% of hypertensive individuals are aware of their condition, only 53.6% are under treatment, and nearly 75% fail to reach the recommended target pressure of 140/90 mm Hg. Significantly, only 40 to 50% of hypertensives will be controlled on a single agent, while most patients with more severe hypertension will require 3 or even 4 agents. This article reviews the overall approach to the hypertensive patient, with special emphasis on target blood pressures in special populations and problems frequently encountered in the older patient. PMID- 12545675 TI - Members. American Ophthalmological Society. PMID- 12545674 TI - The matter of wills. Can your cognitively impaired older patient execute a new will? PMID- 12545676 TI - Long-term risk of local failure after proton therapy for choroidal/ciliary body melanoma. AB - PURPOSE: To quantitate long-term risk of local treatment failure after proton irradiation of choroidal/ciliary body melanomas and to evaluate risk of metastasis-related deaths after local failure. METHODS: We followed prospectively 1,922 patients treated at the Harvard Cyclotron between January 1975 and December 1996 for local recurrences of their tumors. Mortality surveillance was completed through June 1999. For analysis, patient follow-up continued until tumor regrowth was detected or, in patients without recurrence, until the date of the last dilated examination prior to April 1998. Actuarial methods were used to calculate rates of recurrence and metastatic deaths. Cox regression models were constructed to evaluate risk factors for these outcomes. RESULTS: Median ocular follow-up after irradiation was 5.2 years. Local recurrence was documented in 45 patients by ultrasound and/or sequential fundus photographs; in 17 more patients, the eye was enucleated due to suspected but unconfirmed tumor growth. Recurrences were documented between 2 months and 11.3 years after irradiation. The 5- and 10-year rates of regrowth, including suspected cases, were 3.2% (95% confidence interval [CI], 2.5%-4.2%), and 4.3% (95% CI, 3.3%-5.6%). Among the 45 documented recurrences, about one half (21) occurred at the margin, presumably due to treatment planning errors. The remaining cases represented extrascleral extensions (nine cases), ring melanomas (six cases), or uncontrolled tumor (nine cases). Recurrence of the tumor was independently related to risk of tumor related death. CONCLUSION: These data, based on relatively long-term follow-up, demonstrate that excellent local control is maintained after proton therapy and that patients with recurrences experience poorer survival. PMID- 12545677 TI - Suturing technique for control of postkeratoplasty astigmatism and myopia. AB - PURPOSE: We previously demonstrated that selective suture removal reduces keratoplasty astigmatism; however, a myopic shift was induced with increasing number of interrupted sutures removed. This study is an attempt to determine the effects of a modified surgical technique on postkeratoplasty myopia, astigmatism, and anisometropia. METHODS: Optical penetrating keratoplasties were performed on 92 eyes of 84 patients. The study group consisted of 92 consecutive penetrating keratoplasties performed using 12 interrupted 10-0 nylon sutures and a tight 12 bite continuous suture, and use of an average keratometry (K) reading of 46.00 diopters for eyes undergoing combined and intraocular lens (IOL) exchange procedures. All patients had refraction, keratometry, and videokeratoscopy postoperatively, starting at 6 weeks and at the completion of selective suture removal. RESULTS: Prior to suture removal, the average spherical equivalent was 0.160 +/- 3.59 diopters. It was -1.58 +/- 3.66 diopters at the completion of suture removal at 1 year and -1.44 +/- 3.72 at the last follow-up visit, averaging 20.7 months. Final residual refractive, keratometric, and videokeratoscopic astigmatism was 2.81 +/- 1.82, 4.19 +/- 2.94, and 3.58 +/- 2.03 diopters, respectively. Anisometropia, using the spherical equivalent of the operated and fellow eyes, was 2.49 +/- 2.25 diopters at completion of the study. A best corrected visual acuity of 20/50 or better was achieved in 50 patients (59%). CONCLUSIONS: Low myopic spherical equivalent refraction and anisometropia with moderate residual astigmatism were achieved by using tighter continuous sutures, an average K reading of 46 diopters for calculation of IOL power, and selective removal of fewer sutures. PMID- 12545678 TI - The dehiscent Hughes flap: outcomes and implications. AB - PURPOSE: The modified Hughes procedure is used to reconstruct full-thickness lower eyelid defects. A tarsoconjunctival flap from the upper eyelid replaces the posterior lamella, whereas a skin graft, a skin flap, or a skin-muscle flap restores the anterior lamella. The conjunctival pedicle from the upper eyelid is divided after vascularization of the reconstructed lower eyelid is judged to be adequate (traditionally, at least 3 weeks postoperatively). This study reviews the outcomes of patients in whom the conjunctival flap prematurely dehisced. METHODS: Eight patients were identified during a 15-year interval. The posterior lamellar defects ranged in size from 13 to 30 mm horizontally and 5 to 8 mm vertically. The average age at the time of eyelid reconstruction was 72 years (range, 60-84 years). Flap dehiscence, resulting in each case from accidental trauma, occurred between 1 and 11 days postoperatively. Surgical repair of the dehiscence was unsuccessfully attempted in one case; otherwise, the eyelids were permitted to heal spontaneously with the application of erythromycin ophthalmic ointment as the sole therapy. RESULTS: Although the result was satisfactory in each case, one patient, who had dry eyes from Sjogren's syndrome, required secondary surgery to treat mild lagophthalmos and lower eyelid retraction. Follow up ranged from 3 to 122 months (median, 6.5 months). CONCLUSIONS: The ultimate functional and aesthetic outcomes after premature, traumatic dehiscence of a Hughes flap were surprisingly good, suggesting that elective division of the conjunctival pedicle in routine cases can be performed relatively soon after the primary reconstructive procedure. PMID- 12545679 TI - Endophthalmitis in patients with disseminated fungal disease. AB - BACKGROUND/PURPOSE: Fungal endophthalmitis caused by dissemination from extraocular fungal infections has been reported to vary between 9% and 45%. However, recent clinical experience disagrees with that. This study is an investigation of patients in an inner city teaching hospital, the risks associated with endogenous fungal endophthalmitis, and this incidence. METHODS: All ophthalmology consultations between February 1995 and August 2000 that might be associated with disseminated fungal infection were examined in a prospective manner. Patients were excluded if there was no evidence of a positive fungal culture from any site at any time. Visual symptoms were recorded along with ophthalmologic and systemic examination features. Information was gathered, including the identity of cultured organisms, the sites from which the organisms were obtained, and the patients' disposition. RESULTS: During this interval, 170 consultation requests contained the words "endophthalmitis" or "retinitis" and/or indicated concern about disseminated fungal infections. Extraocular fungal infections were found in 114 patients, but only 82 of them had evidence of systemic dissemination. Some patients had more than one organism. The following are listed in decreasing frequency of occurrence: Candida albicans, Torulopsis glabrata, Candida tropicalis, Candida parapsilosis, Candida krusei, Aspergillus niger, and others. Only two patients had evidence of chorioretinitis and progressed to fungal endophthalmitis. CONCLUSIONS: Endophthalmitis was rare among these patients with known fungal infections. Less than 2% had any related ophthalmic manifestations. Nevertheless, since treatment can save vision, evidence of intraocular infection should be sought as eagerly as before. PMID- 12545680 TI - Tetrathiomolybdate as an antiangiogenesis therapy for subfoveal choroidal neovascularization secondary to age-related macular degeneration. AB - PURPOSE: Since previous studies have shown that angiogenesis requires copper, this study assessed the efficacy and safety of oral tetrathiomolybdate, an antiangiogenesis drug that binds copper, in subfoveal choroidal neovascularization (CNV) secondary to age-related macular degeneration. METHODS: This phase I trial involved 10 patients with age-related active subfoveal CNV. After patient consent was obtained and initial laboratory tests were performed, patients were given a loading dose of tetrathiomolybdate, followed by a maintenance dose to maintain serum ceruloplasmin (Cp) levels at 5 to 15 mg/dL. Serum Cp levels are a surrogate marker of copper status. Patient follow-up consisted of a detailed protocol that included best corrected visual acuity, measurement of extent of CNV (both classic and occult) on fluorescein angiograms, and laboratory tests to ensure that anemia did not develop. The study was approved by the institutional review board of the University of Michigan Medical Center and by the Food and Drug Administration. RESULTS: Follow-up of the 10 patients ranged from 4 to 12 months. The targeted serum Cp level was achieved in 8 of the 10 patients. Initially, patients showed stabilization of CNV, but with continued follow-up, all patients showed progression of CNV and loss of visual acuity. Initial mean visual acuity was 20/60; final mean visual acuity was 20/131. At completion of the study, 2 patients showed about a 25% increase in CNV, 1 patient a 60% increase, 1 patient a 100% increase, and 6 patients a 700% to 1,600% increase in CNV. CONCLUSION: At the dosages used in this study, tetrathiomolybdate was ineffective in preventing the progression of CNV secondary to age-related macular degeneration. PMID- 12545681 TI - Surgical approaches to the management of epithelial cysts. AB - PURPOSE: The purpose of this study was to review management strategies for treatment of epithelial cysts. STUDY DESIGN: Retrospective consecutive interventional case series. METHODS: Charts of patients treated for epithelial ingrowth over a 10-year period by a single surgeon (J.A.H.) were reviewed. Cases of epithelial cysts were identified and the following data were recorded: details of ocular history, preoperative and postoperative visual acuity, intraocular pressure (IOP), ocular examination findings, type of surgical intervention, and details of subsequent procedures performed. RESULTS: Seven eyes with epithelial cysts were identified. Patients ranged in age from 1 1/2 years to 53 years at presentation. Three patients were children. Four cysts were due to trauma, one was presumably congenital, one developed after corneal perforation in an eye with Terrien's marginal degeneration, and one developed after penetrating keratoplasty. Three patients were treated with vitrectomy, en bloc resection of the cyst and associated tissue, fluid-air exchange, and cryotherapy. Four patients were treated with conservative strategy consisting of cyst aspiration (three cases) or local excision (one "keratin pearl" cyst) and endolaser photocoagulation of the collapsed cyst wall or base. In all cases, the epithelial tissue was successfully eradicated; one case required a second excision (follow up, 9 months to 78 months; mean, 45 months). Two eyes required subsequent surgery for elevated IOP, two for cataract extraction, and one for a second penetrating keratoplasty. Final visual acuity ranged from 20/20 to hand motions, depending on associated ocular damage. Best visual results were obtained in the more conservatively managed eyes. CONCLUSION: Epithelial cysts can be managed conservatively in selected patients with good results. This strategy may be particularly useful in children, in whom preservation of the lens, iris, and other structures may facilitate amblyopia management. PMID- 12545682 TI - Multicenter prospective, randomized, double-masked, placebo-controlled study of Rheopheresis to treat nonexudative age-related macular degeneration: interim analysis. AB - OBJECTIVE: To evaluate the safety and efficacy of Rheopheresis blood filtration to treat intermediate- to late-stage preangiogenic age-related macular degeneration (AMD) with soft drusen. DESIGN: Multicenter, prospective, randomized, double-masked, placebo-controlled clinical trial. PARTICIPANTS: First 43 randomized patients (28 Rheopheresis and 15 placebo-control patients) with available baseline and 3-month postbaseline best corrected visual acuity (BCVA) measurements and intermediate- to late-stage preangiogenic AMD with multiple large soft drusen and elevated serum levels of targeted macromolecules. INTERVENTION: Patients were randomly assigned to receive eight Rheopheresis or eight placebo procedures over 10 weeks. MAIN OUTCOME MEASURES: ETDRS BCVA measurements at baseline, 3, 6, 9, and 12 months postbaseline. RESULTS: In primary eyes, the mean LogMAR line difference between Rheopheresis and placebo control eyes was 1.6 lines at 12 months postbaseline; the difference was significant throughout the first posttreatment year (P = .0011, repeated measures analysis). Thirteen percent of Rheopheresis compared with 0% of placebo-control eyes had a > or = 3-line improvement in BCVA at 12 months postbaseline. Four percent of Rheopheresis compared with 18% of placebo-control eyes had a > or = 3 line loss in BCVA. The subgroup of patients whose primary eyes had baseline BCVA worse than 20/40 demonstrated a mean LogMAR difference between Rheopheresis and placebo-control eyes equaling 3.0 lines at 12 months postbaseline; the difference was significant throughout the first posttreatment year (P = .0014, repeated measures analysis). Sixteen percent of Rheopheresis compared with 0% of the placebo-control eyes had a > or = 3-line improvement in BCVA at 12 months postbaseline. Five percent of Rheopheresis compared with 29% of placebo-control eyes had a > or = 3-line loss in BCVA. Fifty-eight percent of Rheopheresis eyes improved to 20/40 or better, compared with 14% of placebo-control eyes. No serious treatment-related adverse events were observed. CONCLUSIONS: Rheopheresis demonstrated statistically significant and clinically relevant effects on BCVA when compared with placebo controls for the 12-month study interval. Untreated patients with BCVA worse than 20/40 with intermediate- to late-stage preangiogenic AMD, soft drusen, and elevated blood factors were at risk for substantial visual loss. A sample size larger than 43 patients is important to provide a basis for widespread adoption of novel therapeutic options for AMD such as Rheopheresis. Therefore, enrollment to 150 patients is continuing. PMID- 12545683 TI - Projected impact of travoprost versus both timolol and latanoprost on visual field deficit progression and costs among black glaucoma subjects. AB - PURPOSE: We compared differences associated with use of travoprost and latanoprost on both progression of perimetric loss over time and associated costs among black patients. METHODS: Patients with primary open-angle glaucome or ocular hypertension were randomly assigned to one of four arms in a 12-month, double-masked study: travoprost (0.004% or 0.0015%), latanoprost (0.005%), or timolol (0.5%). Forty-nine patients received 0.004% travoprost, 43 received latanoprost, and 40 received timolol. We applied algorithms found in published studies that link intraocular pressure (IOP) control to visual field progression and calculated the likelihood of visual field deterioration based on IOP data. This was used to estimate differences in medical care costs. RESULTS: The average IOP was lower for patients receiving travoprost than for patients receiving latanoprost or timolol (17.3 versus 18.7 versus 20.5 mm Hg respectively, P < .05). Travoprost-treated patients had a smaller predicted change in visual field defect score (VFDS) than latanoprost-treated patients and timolol-treated patients, and significantly fewer were expected to demonstrate visual field progression. Medical care costs would be higher for latanoprost-treated and timolol-treated patients. CONCLUSIONS: Recent studies have provided algorithms linking IOP control to changes in visual fields. We found that treatment with travoprost was associated with less visual field progression and potential cost savings. PMID- 12545684 TI - Activated satellite cells are present in uninjured extraocular muscles of mature mice. AB - PURPOSE: We recently demonstrated that there is a continuous process of myonuclear addition into normal, uninjured adult myofibers in rabbit extraocular muscles (EOM). This phenomenon is not seen in skeletal muscles from normal, adult limbs. These features may explain the selective involvement of the EOM in progressive external ophthalmoplegia and oculopharyngeal muscular dystrophy due to an accumulation of damaged DNA in mitochondria and nuclei within the EOM as a result of repeated cycling of the muscle satellite cells. Many testable hypotheses flow from these observations. We investigated whether continuous myonuclear addition is present in normal mouse EOM so that mouse models of genetic disorders can be used to study the pathogenic mechanisms and to test potential therapies for human muscle disorders. METHODS: Bromodeoxyuridine (brdU) was injected intraperitoneally into C57 adult mice every 2 hours for 12 hours. Twenty-four hours later the animals were sacrificed, and the globes with the muscles attached were prepared for immunohistochemical localization of brdU positive nuclei within the EOM. All cross sections were immunostained for both brdU and either dystrophin or laminin. RESULTS: All the rectus muscles from the mouse EOM examined contained both satellite cells and myonuclei that were positive for brdU. This demonstrates the division of satellite cells and the fusion of their daughter cells with existing adult EOM myofibers in mice. CONCLUSIONS: These data indicate that the process of continuous myonuclear addition is also active in mouse EOM. These findings will allow various mutant mouse models to be used to study the pathogenesis and treatment of various muscle disorders. The existence of continuous myonuclear addition in adult, uninjured EOM fundamentally changes the accepted notion that EOM myofibers are postmitotic. PMID- 12545685 TI - Toxicity and dose-response studies of 1 alpha-hydroxyvitamin D2 in LH beta-Tag transgenic mice. AB - PURPOSE: The study objective is to determine the effectiveness of a vitamin D analogue, 1 alpha-hydroxyvitamin D2 (1 alpha-OH-D2), in inhibiting retinoblastoma in a transgenic retinoblastoma model (LH beta-Tag mouse) and to evaluate its toxicity. Previous studies of 1 alpha-OH-D2 in athymic mice with human retinoblastoma xenografts suggested efficacy in tumor suppression and suitability for human treatment. METHODS: LH beta-Tag mice (N = 142), 8 to 10 weeks old, were randomly assigned to treatment groups receiving either control (vehicle) or 0.1, 0.3, 0.5, or 1.0 microgram/day of 1 alpha-OH-D2 via oral gavage five times a week for 5 weeks. Animals were then euthanized. The eyes were enucleated, processed histologically, and serially sectioned. Three sections of each eye were microscopically examined, and mean tumor area was measured using Optimus software. Toxicity was assessed by mortality, weight loss, serum calcium levels, and kidney calcification. RESULTS: The mean tumor size in each 1 alpha-OH-D2 group was smaller than in controls (P values < .02): control, 90,248 microns 2; 0.1 microgram, 31,545 microns 2; 0.3 microgram, 16,750 microns 2; 0.5 microgram, 30,245 microns 2; and 1.0 microgram, 16,049 microns 2. No dose-dependent response curve was evident. Mortality was higher in the groups receiving the 0.5 microgram and 1.0 microgram doses (P values < .01) than in the other treatment groups and the control group. CONCLUSION: In the LH beta-Tag mouse, 1 alpha-OH-D2 inhibits retinoblastoma with no increased mortality at lower doses (0.1 to 0.3 microgram). 1 alpha-OH-D2 has been approved by the Food and Drug Administration as an investigative drug for cancer treatment and has shown efficacy with low levels of toxicity in adult cancer trials. 1 alpha-OH-D2 meets the criteria for human clinical trials. PMID- 12545686 TI - Clinical decision making based on data from GDx: one-year observations. AB - PURPOSE: To determine whether information derived from the GDx scanning laser polarimeter aids in the clinical decision-making process for patients with various types of glaucoma. METHODS: Over a 4-month period, 342 consecutive patients with primary open-angle glaucoma, ocular hypertension, angle-closure glaucoma, or secondary glaucomas or in whom the diagnosis of glaucoma was uncertain were evaluated with the GDx scanning laser. After 1 year, 153 patients with glaucoma underwent GDx analysis again. Chart review revealed that 42 of the 153 patients had a change in therapy as a result of the GDx evaluation combined with analysis of visual fields, optic disc cupping, and intraocular pressure (IOP). Outcomes were then compared. RESULTS: The group who had a change in therapy had a higher average GDx number (51.5 +/- 26.1 vs 37.0 +/- 23.5 [P = .001]) at the initial visit and higher IOP (18.2 +/- 4.6 vs 16.0 +/- 3.2 mm Hg [P = .005]). In spite of a change in therapy, at an average of 344 days later, IOP was unchanged (18.3 +/- 5.3 vs 15.7 +/- 3.2 mm Hg [P = .001]) and GDx values in the altered therapy group were higher than at baseline (57.3 +/- 27.9 vs 36.7 +/- 23.4 [P = .001]), although the differences within each group did not achieve statistical significance. CONCLUSION: GDx analysis may be helpful in determining patients at risk for damage from glaucoma, even in eyes in which cup-disc ratio and field loss have not progressed. Changing medications without significantly reducing IOP may be insufficient to halt increases in GDx numbers and may indicate a need for more aggressive therapy. PMID- 12545687 TI - Bacterial resistance after short-term exposure to antibiotics. AB - PURPOSE: To determine if there is a difference in antibiotic sensitivity to coagulase-negative Staphylococcus (CNS) cultured from the host versus the donor cornea at the time of corneal transplantation. Then to apply this knowledge to preoperative preparation of patients undergoing eye surgery. METHOD: A total of 923 donor corneas stored in Optisol and 895 host corneas with no preoperative antibiotic exposure were cultured. Forty-two CNS positive cultures grew from the donor corneas and 40 from the host corneas (P = .5). RESULTS: There was an increase in resistance in the bacteria cultured from the donor compared with the host. The most striking changes occurred in host versus donor to: ciprofloxacin 27.5% (P = .0033); gentamicin 27% (P = .0113); tobramycin 31.6% (P = .059). The combination of polymixin, bacitracin, and neomycin (P/B/N) was significantly better than ciprofloxacin, gentamicin, and tobramycin or the combination of ciprofloxacin, gentamicin, and tobramycin (C/G/T) (P = .0007). CONCLUSION: The combination of C/G/T exhibited the highest change to resistant bacteria. P/B/N was the most effective commercially available preparation. These results should be considered when making the decision about which preoperative antibiotic to use, if any. PMID- 12545689 TI - Vitreous penetration of orally administered gatifloxacin in humans. AB - PURPOSE: To investigate the penetration of gatifloxacin, a novel extended spectrum fluoroquinolone antibiotic, into the vitreous humor after oral administration. METHODS: A prospective, nonrandomized clinical study of 20 consecutive patients scheduled for pars plana vitrectomy surgery between September 2001 and February 2002 at the Cullen Eye Institute, Houston, Texas. Aqueous, vitreous, and serum samples were obtained and analyzed from 20 patients after oral administration of two 400-mg gatifloxacin tablets taken 12 hours apart before surgery. Assays were performed using high-performance liquid chromatography. RESULTS: Mean gatifloxacin concentrations in serum (n = 19), vitreous (n = 19), and aqueous (n = 10) 4.98 +/- 1.14 micrograms/mL, 1.35 +/- 0.36 microgram/mL, and 1.09 +/- 0.57 micrograms/mL, respectively. Mean sampling times after oral administration of the second gatifloxacin tablet for serum, vitreous, and aqueous were 2.99 +/- 0.73 hours, 3.79 +/- 0.81 hours, and 3.71 +/- 0.87 hours, respectively. The percentages of serum gatifloxacin concentration achieved in the vitreous and aqueous were 27.13% and 21.85%, respectively. Mean inhibitory vitreous and aqueous MIC90 levels were achieved against a wide spectrum of pathogens, including Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Propionibacterium acnes, Haemophilus influenzae, Escherichia coli, Bacillus cereus, Neisseria gonorrhoeae, Proteus mirabilis, and other organisms. CONCLUSIONS: Gatifloxacin is a novel fourth-generation fluoroquinolone antibiotic that has MIC90 levels significantly lower than those of other fluoroquinolone agents. Furthermore, it penetrates well into the vitreous cavity in the noninflamed eye. Potential uses for oral gatifloxacin may include prophylaxis against endophthalmitis in open globe injuries, surgical prophylaxis against postoperative endophthalmitis, and adjunctive therapy for the current management of bacterial endophthalmitis. PMID- 12545688 TI - Long-term analysis of LASIK for the correction of refractive errors after penetrating keratoplasty. AB - PURPOSE: To determine the long-term safety and effectiveness of laser-assisted in situ keratomileusis (LASIK) in the treatment of refractive errors following penetrating keratoplasty. METHODS: A retrospective review was done of 57 eyes of 48 patients with anisometropia or high astigmatism who were unable to wear glasses or a contact lens after penetrating keratoplasty and who underwent LASIK for visual rehabilitation. Uncorrected visual acuity (UCVA), best spectacle corrected visual acuity (BCVA), and corneal transplant integrity were recorded before surgery as well as up to 60 months after LASIK. RESULTS: The mean follow up after the LASIK was 21.4 +/- 14.2 months (range, 3-60 months). Mean preoperative spherical equivalent (SE) was -4.19 +/- 3.38 diopters (D). Mean preoperative astigmatism was 4.67 +/- 2.18 D. Preoperative BCVA was 20/40 or better in 42 eyes (74%). At 2 years the mean SE was -0.61 +/- 1.81 D and mean astigmatism was 1.94 +/- 1.35 D for the 28 eyes with follow-up. UCVA was 20/40 or better in 12 eyes (43%), and BCVA was 20/40 or better in 24 eyes (86%) at 2 years. A gain in BCVA of one line or more was seen in eight eyes (29%). Two eyes (7%) had loss of two or more lines of BCVA at 2 years. Nine eyes (16%) developed epithelial ingrowth. Five eyes (9%) in this series had repeat corneal transplants. CONCLUSIONS: LASIK is effective for reducing ametropia after penetrating keratoplasty. Proper patient counseling is necessary because the results of LASIK after penetrating keratoplasty are not as good as, and complications are more frequent than, in eyes with naturally occurring myopia and astigmatism. Complications are especially common in patients with mismatch of the donor and host cornea and in those with poor endothelial cell function. PMID- 12545690 TI - Assessment of the retinal nerve fiber layer of the normal and glaucomatous monkey with scanning laser polarimetry. AB - PURPOSE: To describe and test a method for assessment of the monkey retinal nerve fiber layer (RNFL) with scanning laser polarimetry. METHODS: A scanning laser polarimeter was modified to accommodate a variable corneal polarization compensator. Corneal polarization magnitude (CPM) and corneal polarization axis (CPA) of the anterior segment birefringence of normal and glaucomatous cynomolgus monkey eyes were determined from a polarimetry image of the Henle fiber layer. Next, the variable compensator was adjusted to minimize the anterior segment birefringence. RNFL measurements were then obtained. All images were compared with simultaneous optic disc stereoscopic photographs. RESULTS: CPM was small in each of the eyes examined, ranging from 5.7 nm to 9.0 nm. CPA ranged from -62 degrees to 79 degrees. (Nasally upward CPA values were recorded as negative; nasally downward CPA values were recorded as positive.) When eye-specific compensation was used, RNFL retardation profiles mimicked the expected appearance of the RNFL in all eyes. We also observed a substantial decrease in retardation in eyes with experimental glaucoma compared with healthy fellow eyes. CONCLUSIONS: Individualized anterior segment compensation can be achieved in the monkey eye so that the measured birefringence appears to largely reflect the birefringence of the RNFL. Observed differences in retardation between healthy eyes and eyes with experimental glaucoma suggest that scanning laser polarimetry may be useful for detecting and monitoring RNFL loss in experimental primate glaucoma. PMID- 12545691 TI - A very large Brazilian pedigree with 11778 Leber's hereditary optic neuropathy. AB - PURPOSE: We conducted extensive epidemiological, neuro-ophthalmological, psychophysical, and blood examinations on a newly discovered, very large pedigree with molecular analysis showing mtDNA mutation for Leber's hereditary optic neuropathy (LHON). METHODS: Four patients representing four index cases from a remote area of Brazil were sent to Sao Paulo, where complete ophthalmological examinations strongly suggested LHON. Molecular analysis of their blood demonstrated that they were LHON, homoplasmic 11778, J-haplogroup. They had an extensive family that all lived in one rural area in Brazil. To investigate this family, we drew on a number of international experts to form a team that traveled to Brazil. This field team also included several members of the Federal University of Sao Paulo, and together we evaluated 273 of the 295 family members that were still alive. We conducted epidemiological interviews emphasizing possible environmental risk factors, comprehensive neuro-ophthalmological examinations, psychophysical tests, Humphrey visual field studies, fundus photography, and blood testing for both mitochondrial genetic analysis and nuclear gene linkage analysis. RESULTS: The person representing the first generation case immigrated from Verona, Italy, to Colatina. Subsequent generations demonstrated penetrance rates of 71%, 60%, 34%, 15%, and 9%. The percentages of males were 60%, 50%, 64%, 100%, and 100%. Age at onset varied from 10 to 64 years, and current visual acuities varied from LP to 20/400. CONCLUSIONS: Almost 95% of a nearly 300-member pedigree with LHON 11778 were comprehensively studied. Analysis of environmental risk factors and a nuclear modifying factor from this group may help address the perplexing mystery of LHON: Why do only some of the genetically affected individuals manifest the disease? This fully described database may also provide an excellent opportunity for future clinical trials of any purported neuroprotective agent. PMID- 12545692 TI - The disc damage likelihood scale: reproducibility of a new method of estimating the amount of optic nerve damage caused by glaucoma. AB - PURPOSE: The major objective of this study was to test the reproducibility of a new method of estimating the amount of optic disc damage in patients with glaucoma. METHODS: The Disc Damage Likelihood Scale (DDLS) is based on the appearance of the neuroretinal rim of the optic disc corrected for disc diameter. The eight stages, extending from no damage to far advanced damage, are based on the width of the neuroretinal rim or the circumferential extent of absence of neuroretinal rim. Reproducibility was measured by two masked observers staging 48 optic nerve stereoscopic photographs by two different methods, the cup/disc ratio (c/d) and the DDLS. Also, reproducibility was assessed by three observers examining 34 eyes of 24 patients. RESULTS: With regard to the photographs, the intraobserver and interobserver reproducibility was better using the DDLS than the c/d ratio (98% versus 85% for intraobserver of reproducibility, and 85% versus 74% for interobserver reproducibility). The DDLS correlated better with the Humphrey Visual Field than did any Heidelberg Retina Tomograph parameter. CONCLUSION: In a clinical setting, the DDLS is as reproducible as, or more reproducible than, the c/d ratio system of estimating the amount of disc damage in patients with glaucoma. PMID- 12545693 TI - Evolution of the tapetum. AB - PURPOSE: To review, contrast, and compare current known tapetal mechanisms and review the implications for the evolution of the tapetum. METHODS: Ocular specimens of representative fish in key piscine families, including Acipenseridae, Cyprinidae, Chacidae; the reptilian family Crocodylidae; the mammalian family Felidae; and the Lepidopteran family Sphingidae were reviewed and compared histologically. All known varieties of tapeta were examined and classified and compared to the known cladogram representing the evolution of each specific family. RESULTS: Types of tapeta include tapetum cellulosum, tapetum fibrosum, retinal tapetum, invertebrate pigmented tapetum, and invertebrate thin film tapetum. All but the invertebrate pigmented tapetum were examined histologically. Review of the evolutionary cladogram and comparison with known tapeta suggest that the tapetum evolved in the Devonian period 345 to 395 million years ago. Tapeta developed independently in at least three separate orders in invertebrates and vertebrates, and yet all have surprisingly similar mechanisms of light reflection, including thin-film interference, diffusely reflecting tapeta, Mie scattering, Rayleigh scattering, and perhaps orthogonal retroreflection. CONCLUSION: Tapeta are found in invertebrates and vertebrates and display different physical mechanisms of reflection. Each tapetum reflects the wavelengths most relevant to the species' ecological niche. With this work, we have hypothesized that the tapetum evolved independently in both invertebrates and vertebrates as early as the Devonian period and coincided with an explosion of life forms. PMID- 12545695 TI - Delta-9-tetrahydrocannabinol (THC) in the treatment of end-stage open-angle glaucoma. AB - PURPOSE: Evidence exists that the administration of cannabinoid derivatives can lower intraocular pressure. Some patients with glaucoma believe they are being deprived of a potentially beneficial treatment. Therefore, the Research Advisory Panel of California instituted the Cannabis Therapeutic Research Program to permit compassionate access to cannabinoid derivatives. Data about the potential therapeutic usefulness and toxicity of these agents were collected. This study reviews the results of this program with the specific aim of providing further direction for these investigational efforts. METHODS: A survey of local ophthalmologists indicated an impressive interest in participating in and contributing patients with glaucoma unresponsive to treatment to this study. Appropriate patients were treated with either orally administered delta-9 tetrahydrocannabinol capsules or inhaled marijuana in addition to their existing therapeutic regimen. RESULTS: Although 20 ophthalmologists were approved as investigators, only nine patients were enrolled in the study. An initial decrease in intraocular pressure was observed in all patients, and the investigator's therapeutic goal was met in four of the nine patients. However, the decreases in intraocular pressure were not sustained, and all patients elected to discontinue treatment within 1 to 9 months for various reasons. CONCLUSIONS: This uncontrolled, unmasked, nonrandomized study does not permit definitive conclusions about the efficacy or toxicity of cannabinoids in the treatment of glaucoma. There is an impression that this treatment can lower intraocular pressure, but the development of tolerance and significant systemic toxicity appears to limit the usefulness of this potential treatment. Both patients and ophthalmologists greatly appreciated the opportunity to participate in this study. PMID- 12545696 TI - Diagnostic transvitreal fine-needle aspiration biopsy of small melanocytic choroidal tumors in nevus versus melanoma category. AB - PURPOSE: To report an experience with fine-needle aspiration biopsy of selected small melanocytic choroidal tumors during the interval from April 13, 1983, through January 19, 2001. METHODS: Retrospective descriptive case series report of 34 patients with a small melanocytic choroidal tumor (maximal diameter, < or = 10 mm; thickness, > or = 1.5 mm but < or = 3 mm) evaluated diagnostically by transvitreal fine-needle aspiration biopsy prior to treatment. None of the tumors had invasive features at the time of biopsy. RESULTS: Patients ranged in age from 26 to 73 years (mean, 50.9 years). The evaluated choroidal tumors had a mean maximal basal diameter of 8.0 mm and a mean maximal thickness of 2.4 mm. Eighteen of the 34 tumors (52.9%) had been documented to enlarge prior to biopsy. Biopsy was performed in all cases using a 25-gauge hollow lumen needle and a transvitreal approach via a pars plana puncture site. The biopsy yielded a sufficient aspirate for cytodiagnosis in 22 of 34 cases (64.7%). In these cases, the tumor was classified as malignant melanoma in 16 (47.1% of total), intermediate lesion in 4 (11.8%), and benign nevus in 2 (5.9%). The 12 tumors that yielded an insufficient aspirate and the four lesions that yielded intermediate cells continued to be classified as "nevus versus melanoma" and were monitored periodically for growth or other changes. Four of the 12 tumors that yielded an insufficient aspirate for cytodiagnosis and all four lesions that yielded intermediate cells were eventually reclassified as small choroidal melanomas and treated. The remaining eight tumors that yielded an insufficient aspirate and the two tumors that yielded benign nevus cells were classified as benign nevi at the most recent follow-up evaluation. CONCLUSIONS: Fine-needle aspiration biopsy showed that a substantial proportion of small melanocytic choroidal tumors likely to be classified clinically as small choroidal melanomas in many centers were in fact benign nevi or intermediate lesions. PMID- 12545694 TI - Angle closure in younger patients. AB - PURPOSE: Angle-closure glaucoma is rare in children and young adults. Only scattered cases associated with specific clinical entities have been reported. We evaluated the findings in patients in our database aged 40 or younger with angle closure. METHODS: Our database was searched for patients with angle closure who were 40 years old or younger. Data recorded included age at initial consultation; age at the time of diagnosis; gender; results of slit-lamp examination, gonioscopy, and ultrasound biomicroscopy (from 1993 onward); clinical diagnosis; and therapy. Patients with previous incisional surgery were excluded, as were patients with anterior chamber proliferative mechanisms leading to angle closure. RESULTS: Sixty-seven patients (49 females, 18 males) met entry criteria. Mean age (+/- SD) at the time of consultation was 34.4 +/- 9.4 years (range, 3-68 years). Diagnoses included plateau iris syndrome (35 patients), iridociliary cysts (8 patients), retinopathy of prematurity (7 patients), uveitis (5 patients), isolated nanophthalmos (3 patients), relative pupillary block (2 patients), Weill Marchesani syndrome (3 patients), and 1 patient each with Marfan syndrome, miotic induced angle closure, persistent hyperplastic primary vitreous, and idiopathic lens subluxation. CONCLUSION: The etiology of angle closure in young persons is different from that in the older population and is typically associated with structural or developmental ocular anomalies rather than relative pupillary block. Following laser iridotomy, these eyes should be monitored for recurrent angle closure and the need for additional laser or incisional surgical intervention. PMID- 12545697 TI - The use of antimicrobial peptides in ophthalmology: an experimental study in corneal preservation and the management of bacterial keratitis. AB - PURPOSE: Bacterial keratitis is an ocular infection with the potential to cause significant visual impairment. Increasing patterns of antibiotic resistance have necessitated the development of new antimicrobial agents for use in bacterial keratitis and other serious ocular infections. With a view to exploring the use of novel antimicrobial peptides in the management of ocular infection, we performed a series of experiments using synthetic antimicrobial peptides designed for the eradication of common and serious ophthalmic pathogens. METHODS: Experiments were performed with three clinical ocular isolates--Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis--in three experimental settings: (1) in vitro in a controlled system of 10 mM sodium phosphate buffer, (2) in vitro in modified chondroitin sulfate-based corneal preservation media (Optisol), and (3) in an in vivo animal model (rabbit) simulating bacterial keratitis. In all cases, outcomes were measured by quantitative microbiological techniques. RESULTS: The candidate peptides (CCI A, B, and C and COL-1) produced a total reduction of the test pathogens in phosphate buffered saline. In modified Optisol, the peptides were effective against S epidermidis at all temperatures, demonstrated augmented activity at 23 degrees C against the gram-positive organisms, but were ineffective against P aeruginosa. The addition of EDTA to the medium augmented the killing of P aeruginosa but made no difference in the reduction of gram-positive organisms. In an in vivo rabbit model of Pseudomonas keratitis, COL-1 demonstrated neither clinical nor microbicidal efficacy and appeared to have a very narrow dosage range, outside of which it appeared to be toxic to the ocular surface. CONCLUSION: Our data indicate that the antimicrobial peptides we tested were effective in vitro but not in vivo. In an age of increasing antibiotic resistance, antimicrobial peptides, developed over millions of years as innate defense mechanisms by plants and animals, may have significant potential for development as topical agents for the management of severe bacterial keratitis. However, modifications of the peptides, the drug delivery systems, or both, will be necessary for effective clinical application. PMID- 12545701 TI - 2-deoxy-d-glucose uptake in the inner retina: an in vivo study in the normal rat and following photoreceptor degeneration. AB - PURPOSE: To evaluate, in vivo, at the cellular level, glucose metabolism in the rat inner retina, and to determine how inner retinal glucose metabolism is affected by photoreceptor degeneration. METHODS: Glucose metabolism was evaluated using the 2-deoxyglucose technique. This is an autoradiographic technique that permits evaluation of glucose uptake at the cellular level. The three experimental groups consisted of normal rats (n = 13), dystrophic Royal College of Surgeons rats (n = 3), and rats previously treated with argon green photocoagulation (n = 5). RESULTS: Deoxyglucose uptake in the normal rat was not uniform across the inner retina. Uptake was greatest at the junction of the outer plexiform and inner nuclear layers, and in the inner plexiform layer. Following focal or diffuse photoreceptor loss, there was a marked decrease in the amount of deoxyglucose uptake at the junction of the outer plexiform and inner nuclear layers. CONCLUSION: The pattern of uptake of deoxyglucose in the inner retina is consistent with abundant uptake of deoxyglucose by Muller cells and at sites of synaptic transmission. The decline in deoxyglucose uptake following diffuse or focal photoreceptor loss indicates that there is diminished inner retinal glucose uptake following photoreceptor loss. This change in inner retinal glucose metabolism following photoreceptor loss may help to explain the inner retinal vascular changes observed following photocoagulation and in retinal dystrophies. PMID- 12545698 TI - Human retinal pigment epithelial lysis of extracellular matrix: functional urokinase plasminogen activator receptor, collagenase, and elastase. AB - PURPOSE: To show (1) human retinal pigment epithelial (HRPE) expression of functional urokinase plasminogen activator receptor (uPAR; CD87), (2) HRPE secretion of collagenase and elastase, (3) uPAR-dependent HRPE migration, and (4) uPAR expression in diseased human retinal tissue. METHODS: Immunohistochemistry for uPAR was performed on cultured HRPE cells and in sections of human retina. Double-immunofluorescent staining of live human RPE cells with anti-CR3 antibody (CD11b) was performed to demonstrate the physical proximity of this beta 2 integrin with uPAR and determine whether associations were dependent on RPE confluence and polarity. Extracellular proteolysis by HRPE uPAR was evaluated using fluorescent bodipy-BSA and assessed for specificity by plasminogen activator inhibitor-1 (PAI-1) inhibition. The effect of interleukin-1 beta (IL-1 beta) on uPAR expression was assessed. Collagenase and elastase secretion by unstimulated and IL-1-stimulated HRPE cells was measured by 3H-labelled collagen and elastin cleavage. HRPE-associated collagenase was also assessed by cleavage of fluorescent DQ-collagen and inhibited by phenanthroline. Using an extracellular matrix assay, the roles of uPAR and collagenase in HRPE migration were assessed. RESULTS: Immunoreactive uPAR was detected on cultured HRPE cells and increased by IL-1. On elongated, live HRPE cells, uPAR dissociated from CD11b (CR3) and translocated to anterior poles of migrating cells. Extracellular proteolysis was concentrated at sites of uPAR expression and specifically inhibited by PAI-1. Cultured HRPE cells secreted substantial, functional collagenase and elastase. IL-1 upregulated uPAR, collagenase, and elastase activities. Specific inhibition of uPAR, and to a lesser degree collagenase, reduced HRPE migration in matrix/gel assays. Immunoreactive uPAR was present along the HRPE basolateral membrane in retinal sections and in sections of diseased retinal tissue. CONCLUSIONS: HRPE cells express functional uPAR, collagenase, and elastase, which may permit HRPE proteolysis and migration. uPAR polarization may concentrate proteolysis at the leading edge of migrating HRPE cells. PMID- 12545700 TI - Diabetes-induced dysfunction of retinal Muller cells. AB - PURPOSE: This study tested the hypothesis that the function of the glutamate transporter in retinal Muller cells is compromised early in the course of diabetes by a mechanism involving oxidation. Dysfunction of this transporter, which removes glutamate from the extracellular space, may play a critical role in the disruption of glutamate homeostasis that occurs in the diabetic retina. Because glutamate is toxic to retinal neurons and is likely to exacerbate oxidative stress, elucidation of the mechanisms by which diabetes elevates the concentration of this amino acid may help to better understand the pathogenesis of diabetic retinopathy. METHODS: Muller cells were freshly isolated from normal rats and those made diabetic by streptozotocin injection. The activity of the Muller cell glutamate transporter, which is electrogenic, was monitored via the perforated-patch configuration of the patch-clamp technique. RESULTS: Four weeks after the onset of hyperglycemia, dysfunction of the Muller cell glutamate transporter was detected (P = .005). After 13 weeks of streptozotocin-induced diabetes, the activity of this transporter was decreased by 67% (P = .001). Consistent with oxidation causing this dysfunction, exposure to a disulfide reducing agent rapidly restored the activity of this transporter in Muller cells from diabetic retinas. CONCLUSIONS: Soon after the onset of experimental diabetes, the function of the glutamate transporter in Muller cells is decreased by a mechanism that is likely to involve oxidation. The demonstration that the activity of this transporter can be rapidly restored raises the possibility that targeting this molecule for therapeutic intervention may restore glutamate homeostasis and, thereby, ameliorate sight-threatening complications of diabetic retinopathy. PMID- 12545702 TI - Progression of visual field loss in untreated glaucoma patients and suspects in St Lucia, West Indies. AB - PURPOSE: A 1986-1987 survey found an 8.8% prevalence of open-angle glaucoma in the black population of St Lucia, West Indies. This follow-up study assessed progression of visual field loss in untreated glaucoma patients and persons with suspected glaucoma 10 years later. METHODS: Subjects were 205 patients with or suspected of having glaucoma. The 1987 data included age, sex, visual acuity, and visual fields measured by automated threshold perimetry (Humphrey C-30-2 test). The 1997 data included intraocular pressure, visual acuity, and visual fields measured by the same test. Exclusion criteria included field unreliability, field improvement due to vision improvement, nonglaucomatous vision deterioration, glaucoma treatment since 1988, and scoring of a field as end-stage in 1987. Visual fields were scored by algorithms for the Advanced Glaucoma Intervention Study (AGIS) and Collaborative Initial Glaucoma Treatment Study (CIGTS). RESULTS: By AGIS criteria, 55% of 146 right eyes and 52% of 141 left eyes progressed. In linear regressions, progression severity was unassociated with male or female sex, intraocular pressure, or baseline visual field score, but was positively associated with age (P < .001, right; P = .002, left). By CIGTS criteria, more eyes progressed. The cumulative probability of reaching end-stage disease in 10 years in at least on eye was about 16% by AGIS criteria and was 35% by CIGTS criteria. CONCLUSIONS: These data provide a unique opportunity to study progression of untreated glaucoma. A considerably larger percentage of eyes showed progression of visual field loss, and the rate of progression was greater than in studies of visual field loss in treated eyes. PMID- 12545704 TI - [The molecular mechanism of neuronal migration]. AB - A large amount of work since the late 1800 have shown that, during embryonic and postnatal development, the majority, if not all, neurons in mammalian nervous system have to migrate a certain distance to reach their final destination. An interesting question is how neurons are guided for their migration. We are interested in the molecular mechanisms underlying neuronal migration and our work published in 1999 indicates that there are diffusible molecules in the brain which can guide the direction of migrating neurons; specifically, a secreted protein called Slit is repulsive to neurons and its concentration gradient guides neuronal migration. PMID- 12545703 TI - Analysis of the keratocyte apoptosis, keratocyte proliferation, and myofibroblast transformation responses after photorefractive keratectomy and laser in situ keratomileusis. AB - PURPOSE: To test the hypothesis that (1) there are quantitative differences in the cellular responses in the corneal stroma after photorefractive keratectomy (PRK) for low myopia compared to high myopia and (2) there are both qualitative and quantitative differences in the cellular responses in the corneal stroma after PRK for high myopia and laser in situ keratomileusis (LASIK) for high myopia. METHODS: PRK for low myopia (-4.5 diopters [D]), PRK for high myopia ( 9.0 D), and LASIK for high myopia (-9.0 D) were performed in rabbit eyes, and corneas were obtained for examination at 4, 24, and 72 hours, 1 and 4 weeks, and 3 months after surgery. A total of 144 rabbits were included in the study. Stromal apoptosis, necrosis, mitosis, myofibroblast generation, and inflammatory cell infiltration were evaluated by immunohistochemical methods and electron microscopy. RESULTS: Keratocyte apoptosis/necrosis and the subsequent proliferation and density of myofibroblasts were qualitatively and quantitatively different in PRK for high myopia compared to either PRK for low myopia or LASIK for high myopia. Significant inflammatory cell infiltration was noted in both PRK and LASIK but appeared to be greater in PRK for high myopia. CONCLUSIONS: The qualitative and quantitative differences in the cellular wound healing response after PRK for high and low myopia and LASIK for high myopia are likely determinants of the clinical differences in refractive outcome and some of the complications, such as regression and haze, seen after these procedures. PMID- 12545705 TI - [Systemic aging and replicative senescence: in vivo and in vitro]. AB - Cultured cells lose the ability of DNA synthesis, mitosis, and finally the ability of cell proliferation after they have undergone a finite number of population doublings in vitro, though the cells still maintain the basic metabolic process. This is termed replicative senescence. We review the prevalence of replicative senescence, summarize the features of senescent cells, and then focus on the links between systemic aging and replicative senescence. The present knowledge, albeit still incomplete, proposes that replicative senescence is a reflection of systemic aging at cell level, and it fully confirms replicative senescence as a good model for the research of systemic aging. PMID- 12545699 TI - Retinal pigment epithelial acid lipase activity and lipoprotein receptors: effects of dietary omega-3 fatty acids. AB - PURPOSE: To show that fish oil-derived omega-3 polyunsaturated fatty acids, delivered to the retinal pigment epithelium (RPE) by circulating low-density lipoproteins (LDL), enhance already considerable RPE lysosomal acid lipase activity, providing for more efficient hydrolysis of intralysosomal RPE lipids, an effect that may help prevent development of age-related macular degeneration (ARMD). METHODS: Colorimetric biochemical and histochemical techniques were used to demonstrate RPE acid lipase in situ, in vitro, and after challenge with phagocytic stimuli. Receptor-mediated RPE uptake of fluorescently labeled native, aceto-acetylated, and oxidized LDL was studied in vitro and in vivo. LDL effects on RPE lysosomal enzymes were assessed. Lysosomal enzyme activity was compared in RPE cells from monkeys fed diets rich in fish oil to those from control animals and in cultured RPE cells exposed to sera from these monkeys. RESULTS: RPE acid lipase activity was substantial and comparable to that of mononuclear phagocytes. Acid lipase activity increased significantly following phagocytic challenge with photoreceptor outer segment (POS) membranes. Receptor-mediated RPE uptake of labeled lipoproteins was determined in vitro. Distinctive uptake of labeled lipoproteins occurred in RPE cells and mononuclear phagocytes in vivo. Native LDL enhanced RPE lysosomal enzyme activity. RPE lysosomal enzymes increased significantly in RPE cells from monkeys fed fish oil-rich diets and in cultured RPE cells exposed to their sera. CONCLUSIONS: RPE cells contain substantial acid lipase for efficient metabolism of lipids imbibed by POS phagocytosis and LDL uptake. Diets rich in fish oil-derived omega-3 fatty acids, by enhancing acid lipase, may reduce RPE lipofuscin accumulation, RPE oxidative damage, and the development of ARMD. PMID- 12545706 TI - [Endogenous descending inhibitory/facilitatory system and serotonin (5-HT) modulating spinal nociceptive transmission]. AB - Endogenous descending inhibitory system plays an important role in the modulation of nociceptive transmission. In recent years, descending facilitatory system has been noted. CNS bi-directionally modulates the peripheral nociceptive transmission at the spinal cord level via the descending inhibitory/facilitatory system. Serotonin (5-HT) is the main neurotransmitter in descending pain modulation. Intrathecal application of 5-HT or electrical stimulation or glutamate microinjection into the nucleus raphe magnus (NRM) and nucleus reticularis gigantocellularis(NGC) may produce inhibition and/or facilitation of spinal nociceptive transmission. The contradictory results may be attributed to the nonselective action(s) of 5-HT at multiple subtypes of 5-HT receptor in spinal cord. PMID- 12545707 TI - [Pathophysiological alterations in cultured astrocytes exposed to hypoxia/reoxygenation]. AB - While the research of hypoxic-ischemic injury to the central nervous system has been focused on neurons, astrocytes are also critically involved in and contribute to the hypoxic-ischemic process. The role these cells play appears to be more and more important. There are considerable progress in characterizing the pathophysiological alterations of these cells during hypoxic-ischemic brain damage. PMID- 12545708 TI - [Central control of the hypothalamic-pituitary-adrenocortical axis for stress response]. AB - Stress reaction is an adaptive response of all living organism to stressful events and essential for their survival. The main feature of stress reaction is activation of the hypothalamic-pituitary-adrenocortical (HPA) axis, central control of which is very complex. The hippocampus is involved in integration of sensory information, interpretation of environmental information, and execution of appropriate behavioral and neuroendocrine responses. The amygdala is an executor of stress behavioral, autonomic and neuroendocrine responses. The paraventricular nucleus (PVN) is responsible for initiating HPA axis stress reaction. The negative feedback regulation of stress hormones, local hypothalamic circuits and cytokines probably are involved in regulation of HPA activation. PMID- 12545709 TI - [Studies on learning and memory function-related genes in the hippocampus and the relationship between the cognitive enhancing effect of liuwei dihuang decoction (LW) and gene expression]. AB - Our studies showed that the expressions of the genes including glucocorticoid receptor (GR), mineralocorticoid receptor (MR), bcl-2, c-fos, neural cell adhesion molecule(NCAM), presenilin-2 (PS-2) and apoE in the hippocampus were closely related to the central learning and memory function in senescence accelerated mice (SAM), hydrocortisone(HC)-treated mice and normal mice. The differential display technique was applied to compare mRNAs expression between SAM-prone/8 (SAMP8), and SAM-resistance/1 (SAMR1). Six differentially expressed cDNA bands were identified and two of them are unknown genes. Chronic oral administration of LW (5 g/kg), a traditional Chinese medicinal prescription, significantly ameliorated the deterioration of learning and memory ability in SAMP8 and HC-treated mice and corrected the abnormal expressions of hippocampal genes. Further studies showed that corticosterone significantly affected the gene expression in primary cultured hippocampal neurons. These results suggested that central learning and memory function is closely related to the expressions of hippocampal genes. The imbalance of hypothalamic-pituitary-adrenal (HPA) axis leads to the deterioration of learning and memory function and the abnormal expressions of hippocampal genes. Therefore, one of the important ways of the cognitive enhancing effect of LW is to correct the abnormal expressions of hippocampal genes. PMID- 12545710 TI - [The roles of estrogen and progestin in epileptogenesis and their mechanisms of action]. AB - Clinical data showed that periodic alterations of estrogen and progestin in some women patients with catamenial epilepsy might affect seizure susceptibility. In the present study, in order to explore the roles of estrogen and progestin in epilepsy and elucidate their mechanisms of action, we utilized animal seizure models induced by intracerebroventricular(i.c.v) coriaria lactone(CL) and intraperitoneal(i.p.) bemegride(Be), studied the effects of estrogen and progestin on central nervous system(CNS) functions in behavior, electrophysiological, cellular, molecular and gene levels by means of neuroelectrophysiology, flow cytometry, high performance liquid chromatography, immunocytochemical and in situ hybridization techniques. From our observation we concluded that ovarian hormones are neuroactive steroids and new neuromodulaters, they could affect the excitability of CNS by regulating immediate early genes, amino acid neurotransmitters and N-methyl-D-aspartate receptor(NMDAR). PMID- 12545711 TI - [Actions of insulin-like growth factor-I on heart]. PMID- 12545712 TI - [Neurogenesis in the adult mammalian brain]. PMID- 12545713 TI - [The role of mitogen-activated protein kinase phosphatase in cardiovascular system]. PMID- 12545714 TI - [The role of endostatin in tumor growth and progression of atherosclerosis]. PMID- 12545715 TI - [Proteins involved in regulation of ryanodine receptor]. PMID- 12545716 TI - [Progress in the study of directed differentiation of bone marrow-derived mesenchymal stem cells]. PMID- 12545717 TI - [Nucleocytoplasmic translocation biomacromolecules]. PMID- 12545718 TI - [Adhesion molecules and inflammation following cerebral ischemia]. PMID- 12545719 TI - [Cannabinoids, cannabinoid receptor and immune response]. PMID- 12545720 TI - [The role of cardiomyocyte apoptosis on mechanism of heart failure]. PMID- 12545721 TI - [Vascular endothelial growth factor receptor--structure and function]. PMID- 12545722 TI - [Role of the hypothalamic paraventricular nucleus in cardiovascular regulation]. PMID- 12545723 TI - [Molecular biology of the retinol-binding protein]. PMID- 12545724 TI - [Molecular mechanism of exocytosis in chemical synaptic transmission]. PMID- 12545725 TI - [Central nervous system calcium homeostasis imbalance and brain aging]. AB - Brain aging is associated with a marked decline of memory. The calcium homeostasis imbalance hypothesis postulated that sustained changes in the regulation of intracellular Ca2+ concentration, [Ca2+]i are the major cause of brain aging. Aging may cause the increase of the [Ca2+]i through many mechanisms. These alterations would in turn affect the synaptic transduction, neurotransmitter release and signal transduction, cause memory deficit. This article reviewed the progress about this hypothesis in recent years. PMID- 12545726 TI - [Elementary events and characteristics of calcium signalling]. AB - There are many different elementary intracellular calcium signalling in the cell. These events appear to have a hierarchical organization depending on stimulus intensity. Low levels of stimulation activate individual channels such as the blips or quarks; The next level of organization is the puffs and sparks, which appear to be derived from small clusters of channels. At high stimulus intensities these elementary events are coordinated to give global events. These events not only present the character of Ca2+ release unit, also lead to different complex spatiotemporal organization of calcium signalling, emphasizing the hierarchical organization and local control model of calcium signalling. PMID- 12545727 TI - [Current status of antisense drugs]. AB - The antisense was first imagined as the therapeutic drug at the end of 1970. After 20 years the antisense drugs have been hitting into the market from laboratory and clinical research. Antisense captures general attention with their promise of rational drug design and exquisite specification. But antisense drugs are far more difficult to produce than were originally anticipated. This article reviewed the current situation and present questions involved in their specificity, stability, potency, toxicology, intracellular delivery, administration routes and costs of manufacture, etc. It is likely that the antisense drugs will be a part of the pharmacopoeia in the future and benefit for treatment of human diseases with the solution of present questions. PMID- 12545729 TI - [Metabotropic glutamate receptors and its contribution to the mechanisms of pain]. AB - Metabotropic glutamate receptors (mGluRs) is a novel family of G protein-coupled receptors. Each subtype shows different features in the aspects of molecular biology, neuropharmacology and distribution in the central nervous system. It can be divided into three groups according to the extent of sequence homology. Recent experimental research has demonstrated that mGluRs play an important role in the transmission of pain. This article will review the possible roles of mGluRs in the processing of nociceptive information. PMID- 12545728 TI - [Protein kinase B and its role in the signal transduction pathway mediated by phosphoinositide 3-kinase]. AB - Protein kinase B (PKB) is the expression product of a proto-oncongen (c-akt), which is involved in the signaling pathways initiated by some growth factors and mediated by phosphoinositide 3-kinase (PI3K). PKB is a direct target of PI3K. Similar to many protein kinases, PKB has a specific AH/PH domain which can mediate the interaction between signaling molecules. The lipid second messengers, PI-3, 4-P2 and PI-3,4,5-P3 produced by PI3K, can bind to the AH/PH domain of PKB and of PDK (phosphoinositide dependent protein kinase). This binding translocates PKB and PDK to the plasma membrane, and activates them. PKB is also activated via phosphorylation by PDK and, in turn, will activate the anti-apoptotic machinery, glucose metabolism (glycogen synthesis, glycolysis and glucose uptake) and protein synthesis. All these lead to cell growth and proliferation. PMID- 12545730 TI - [Protective effect of ciliary neurotrophic factor on the hippocampal neuronal damage induced by stress and its mechanisms in rats]. AB - Effect of ciliary neurotrophic factor (CNTF) on behavior and morphology of hippocampal neurons were observed and its mechanisms in rats were explored by Nissl staining, Bielschowsky-Gros-Lawrentjew staining, transmission electron microscopy, behavior determination, primary culture of hippocampal neuron, running photography of living cell, whole-cell patch clamp recording, detection of intracellular free Ca2+ and immunohistochemical detection of P53 protein. The results showed that there was no statistically significant change in the morphology of hippocampal neurons as a result of acute stress. The behavioral activity was increased during acute stress stage, which was not affected by CNTF. In chronic stress stage, neuronal damage in hippocampus was significant, and behavioral activity was significantly decreased under basal line. Administration of CNTF into bilateral hippocampus prevented neurons from damage and improved behavior. In vitro, CNTF could significantly suppress channel current, intracellular Ca2+ content and the expression of P53 protein in the nucleus induced by glutamate. The results suggested that the protective effect of CNTF may involve rapid effects on cell membrane and cytoplasma, and delayed effects on nucleus, thereby improve behavioral defects. PMID- 12545731 TI - [The development of gene chips technology]. PMID- 12545732 TI - [Advances in the study of the effect of nitric oxide on long-term potentiation]. PMID- 12545733 TI - [Progress in the study of leptin receptor]. PMID- 12545734 TI - [Calcineurin-dependent signal pathway: a new signal pathway]. PMID- 12545735 TI - [The mechanism of water retention in congestive heart failure]. PMID- 12545736 TI - [Mechanism of mechanical stimulation sensed by cardiac myocytes]. PMID- 12545737 TI - [Mitogen-activated protein kinases participating cell apoptosis]. PMID- 12545738 TI - [Transcriptional regulation of peripheral nociceptors]. PMID- 12545739 TI - [The effect of vitamin E on immune functions]. PMID- 12545740 TI - [Structure and biological activity of beta amyloid protein precursor]. PMID- 12545741 TI - [Research progress in immune regulation of gonadorelin]. PMID- 12545743 TI - [Human body composition: history, current status and future]. PMID- 12545742 TI - [Progress in the recommended dietary allowance]. PMID- 12545744 TI - [In vivo and in vitro study on the effect of excessive fluoride on type I collagen of rats]. AB - In order to investigate the effects of excessive fluoride on type I collagen in rat bone, rat fluorosis model is induced by giving 221 mg/L NaF in drinking water for two months. The level of serum BGP, inorganic ingredient, the content of collagen and the degree of collagen cross-linking in fluorosis rat bone are determined. Rat calvaria osteoblasts are separated by enzyme digestion. After the treatment of 0.5 and 1.0 mmol/L NaF for 48 hour, effects of excessive fluoride on the expression level of type I collagen of the osteoblasts are detected through immunohistochemical method. The results show that after NaF treatment for two months, the serum BGP and inorganic ingredient content in fluorosis rat bone increase significantly in comparison to those of in control group (P < 0.05). On the contrast, the content of collagen and the degree of collagen cross-linking in fluorosis rat bone decrease significantly (P < 0.05) compared with control group. In in vitro study, the osteoblasts secrete less type I collagen after NaF treatment than they do in control group cells. The results suggest that excessive fluoride can inhibit the synthesis of type I collagen. The decrease of collagen may be one of causal factors to skeletal fluorosis. PMID- 12545745 TI - [Establishment of rat alveolar type II epithelial cell line RAE-1 and its biological characteristics]. AB - The aim is to establish rat alveolar type II epithelial cell line and to provide cell models for epithelial cell cancer research. Highly pure Lewis rat alveolar epithelial cells were successfully isolated by Nycodenz buoyant density gradient centrifugation in a combination with IgG panning method. Semian virus 40 early region large T gene was virally transferred into primary cultures. After neo gene selection, a cell line RAE-1 was established. The characteristics of this cell line were analyzed. Homologous integration of SV40T gene into RAE-1 cell genome was positively detected by PCR, modal chromosome number of RAE-1 cells was 44, and expressions of cytokeratins 8 and 18 at mRNA level were both positively detected by RT-PCR. The establishment of RAE-1 cell line was very important because it could serve as model cells to study epithelial cell neoplastic transformation mechanisms in hazards-caused alveolar epithelial cancer. PMID- 12545746 TI - [Study of the inhibitory effect of PhAsO on gap junctional communication and its damages to cellular DNA of human skin fibroblast cells]. AB - In order to investigate the toxicity of organic arsenic as well as the mechanism of arsenic carcinogenesis, the GJIC between human skin fibroblast cells is detected by the scrape loading/dye transfer assay and single-cell gel electrophoresis(SCGE) assay is used to detect the DNA strand breaks in human skin fibroblast cells exposed to PhAsO. The results show that gap junctional intercellular communication between human skin fibroblast cells is inhibited by PhAsO significantly with a obvious dose-dependent relationship(P < 0.01), suggesting that PhAsO may act as a tumour promoter. PhAsO could induce DNA strand breaks, but the dose-dependent relationship is not observed. The authors suggest that PhAsO may have tumor promoting activity, and further studies are needed. PMID- 12545748 TI - [Compare of distributions of gene polymorphisms about alcohol metabolizing related enzymes in five Chinese nationalities]. AB - Genotypes of alcohol dehydrogenase 2(ADH2) and aldehyde dehydrogenase 2(ALDH2) were detected by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in order to acquire and compare the distributions of polymorphisms about ADH2 and ALDH2 in five Chinese nationalities. The results were as follows: (1) Heterozygous genotype of ADH2 and homozygous genotype of ALDH2 are the two dominant ones in all the five nationalities. (2) The distributions of the two genes 'polymorphisms have certain differences among some nationalities. PMID- 12545747 TI - [Effect of 5 polycyclic aromatic hydrocarbons on estrogen receptor binding in rats]. AB - The estrogenic activity of 5 polycyclic aromatic hydrocarbons(PAHs) of rat uterine wet weight bioassy and estrogen receptor binding bioassy are determined. The results show that penzo(a)pyrene, 7,12-dimethyl benz(a)anthracene and pentacene could statistically increase uterine wet weight of immature female rats(P < 0.01 or P < 0.05) while benz(a)anthracene and chrysene can not significantly increase uterine wet weight(P > 0.05). Penzo(a)pyrene, 7,12 dimethyl benz(a)anthracene and pentacene effectively inhibit the specific 3H estradial binding to rat uterine estrogen receptor(P < 0.01). Whereas benz(a)anthracene and chrysene can not inhibit the specific 3H-estradial binding to rat uterine estrogen receptor(P > 0.05). The results from this study indicate that penzo(a)pyrene, 7,12-dimethyl benz(a) anthracene and pentacene have significant estrogenic activity. PMID- 12545749 TI - [Variations and transformations of nitrogen pollutants in source and tap water of D-lake]. AB - Ammonia, nitrite and nitrate pollutions and variations in source water and tap water of D-Lake are investigated. The removal effects of nitrogen in routine proceeding of water production and the transformations of the different forms of nitrogen in natural condition with simulated test are studied at the same time. The results show that the nitrogen pollutions in source and tap water of D waterworks are higher than T-waterworks', although both waterworks have same resource. Compared with D-waterworks, the nitrogen pollutions of source and tap water in Z-waterworks are lower than D-waterworks' and they have different variation and distribution. The forms of nitrogen could be partly transformed during the routine production, but the total percentage of nitrogen removal efficiency is not significant (just about 20%). The simulate test shows that NH4+(-)N could change into NO2-(-)N, while No2-(-)N into NO3-(-)N, which is partly transformed into nitrogen gas by microorganism and majority of which are remained in tap water by form of NO3-(-)N. PMID- 12545750 TI - [Effect of diet composition on expression of uncoupling protein-1 gene of rat]. AB - In order to explore the effect of different diet composition on uncoupling proteins 1(UCP1) gene expression in brown adipose tissue, male Wistar rats are fed with different diets (basic diet, high protein diet, high fat diet, high calorie diet 1 and 2) for 2 months. The effects of different diets on weight and the ratio of fat mass to body weight are observed. Reverse transcription polymerase chain reaction and western blotting technique are used to measure UCP1 mRNA and protein content of rat brown adipose tissue(BAT). The results show that body weight and the ratio of fat mass to body weight in high calorie 1 and 2 diet groups are obviously higher than those in groups of basic diet, high protein, and high fat diet (P < 0.05), but there is no significant difference among the last three groups (P > 0.05). UCP1 gene expression increases in both high calorie diet 1 group and 2 group, but it is not significantly different among basic diet, high protein and high fat diet groups with equal energy, indicating that energy can induce UCP1 gene expression of rat BAT. PMID- 12545751 TI - [Study of the effects of zinc on bone development applying fetal mouse limbs culture]. AB - The effects of zinc on bone metabolism is explored. A home-made rotating apparatus is used to cultivate fetal mouse front limbs at 16-day gestational age. The activity of alkaline phosphatase(AKP) and histological silce are used to analyze the effects of zinc on bone metabolism. Compared with the control group, the zinc deprived group and Zn2+ 120 mumol/L group have a lower alkaline phosphatase (AKP) activity and the contracted chromatin, membrane rupture and matrix loss are observed in bone cells. The zinc adequate group(Zn2+ concentration are 45 mumol/L and Zn2+ 70 mumol/L respectively) has a higher alkaline phosphatase(AKP) activity, more active bone generation, deeper stain of bone matrix and increased bone trabecula. It is concluded that zinc could affected the bone metabolism and growth; both zinc-deficiency and zinc-excess could alter growth and bone metabolism. PMID- 12545752 TI - [Effects of exogenous nucleotides on the proliferation and migration of intestinal epithelial cells]. AB - The effect of exogenous nucleotides on proliferation and migration of a normal rat small intestinal epithelial cell line IEC-6 is studied. The concentration effects as well as interaction of exogenous nucleotides on proliferation of IEC-6 are measured by MTT. Migration of IEC-6 after wounded is determined by an in vitro model of intestinal epithelial restitution of IEC-6 monolayer. Expression of TGF beta is detected by immunohistochemistry. The results show AMP and GMP remarkably inhibit proliferation of IEC-6 in concentration-dependent manner respectively at 30 mumol/L or 150 mumol/L and more. CMP, UMP and nucleotides mixture can not enhance or inhibit the growth with the exception of inhibition of CMP on proliferation at very high concentration (1440 mumol/L). In contrast, CMP, especially UMP, can remarkably abolish the proliferation-inhibiting effects of AMP or GMP on the cell, when AMP or GMP is supplemented. Nucleotides mixture significantly facilitate migration of IEC-6 after wounded but fail to promote the expression of TGF beta. It is concluded that purine nucleotides inhibits proliferation of IEC-6. Pyrimidine nucleotides can abolish the inhibitive effects of purine nucleotides, and Nucleotides mixture promotes migration of IEC-6 after wounded by a TGF beta-independented way. PMID- 12545753 TI - [Study on the antioxidant effect of Samara oil]. AB - The oil productivity of Samara oil seeds is high and its quality is better than other kinds of oil seeds. Forty Wistar rats (24 month-old) are randomly divided into 4 groups: A (control group), B (vitamin E group), C(Samara oil group) and D(Sea Buckthron oil group). The rats are killed and examined after 28 days of feeding with four different feeds respectively. The results show that compared with the control group, the lipid peroxidation in group B, C, and D decreases while the antioxidant effect increases indicating that the antioxidant effect of Samara oil is strong than that of vitamin E. It is suggested that this natural oil resource might have great potential in the development of food. PMID- 12545754 TI - [Effect of iron on the expression of uncoupling protein 2, 3 gene of obese rat]. AB - In order to study the effect of iron on the expression of UCP2 in white fat and UCP3 in muscle in obese rats, a reverse transcription polymerase chain reaction (RT-PCR) technique is used to measure the expression of UCP2 and UCP3 mRNA. The results show: (1) Adequate iron improves the level of thyroid hormone in blood serum; (2) the body fat content of obese rat in 5, 10, and 20 times of iron supplement groups is lower than that of obese rats in normal iron supplement group; (3) the UCP2 and UCP3 mRNA expression of iron-deficient obese rats is low in iron-deficient obese rats; (4) iron supplement enhances the level of UCP3 mRNA expression in muscle and the level of UCP3 mRNA is the highest in the 5 times iron supplement group and about 2 times of that of basic diet group. However, there is no effect on the UCP2 mRNA expression induced by high energy diet. It suggested that adequate iron can improve the level of serum thyroid hormone, induce the UCP3 mRNA expression in muscle and mobilize fat to produce heat, but had no effect on UCP2 mRNA in white fat of obese rat. PMID- 12545755 TI - [Study on the developmental toxicity of Bisphenol A by using micromass culture in vitro]. AB - The micromass culture of Wistar rat embryo limb bud cells was used to investigate the characteristic of developmental toxicity of Bisphenol A (BPA) and its mechanism in vitro. The results showed that BPA inhibited both proliferation and differentiation of rat limb bud cells in vitro. The high level of BPA appeared to be cytotoxic to Wistar rat embryo limb bud cells in culture and inhibited the clone formation with dose-response relationship. The concentration of BPA for IP50 and ID50 were 38.74 mg/L and 27.93 mg/L respectively. According to Renaults' teratogenic criteria, BPA belonged to a positive teratogen. And the data suggested that the specific inhibition of cell proliferation and differentiation might be one of the mechanisms of high level BPA. PMID- 12545756 TI - [Study on the expired gas of subjects with lactose intolerance by using H2/13CO2 breath test]. AB - The relationship between breath gas and lactose intolerance symptoms is studied in the subjects with lactose intolerance and lactose mal-absorption. The breath gas samples are collected for 6 hours after consumption of 25 g 13C-lactose. Expired H2 concentration and 13CO2 abundance at each detecting time point is measured by gas chromatography and continuous flow Gas Isotope Mass Spectrometry respectively. The cumulative amount of breath H2 and 13CO2 for 6 hours are calculated, and the abdominal symptoms are recorded by the questionnaire for 12 hours. The results show that the H2 peak is significantly higher in the intolerance group than that in the mal-absorption group(P < 0.01). The cumulative amount of breath H2 during 6 hours is significantly higher in the intolerance group than that in the mal-absorption group (P < 0.01). The oral-colon transit time (OCTT) is negatively related to the lactose intolerance symptom scores (r = 0.705). There are no significant differences for the amount of 13CO2 excretion at each detecting time point and cumulative percentage of the expired 13CO2 between the intolerance and the mal-absorption groups. It is suggested that the cumulative breath H2 amount is associated with the severity of lactose intolerance, which can reflect the degree of lactose hydrolysis in the small intestine, while there is no such relationship between the amount of expired 13CO2 and the severity of lactose intolerance. PMID- 12545757 TI - [Detection of the genetically modified organisms in genetically modified soybean and maize by polymerase chain reaction method]. AB - A method for the detection of the (genetically modified organism GMOs) in genetically modified soybean (Round-up Ready soybean, RR soybean) and maize(Bt 176 maize) is described. The polymerase chain reaction (PCR) method is discussed with the genetically modified soybean and maize whose contents are known. The detection limit can be 0.1%, that is to say, we can detect the GMO in the food whose content is only 0.1%, the detection method is just a screening method. The procedure includes: (1) extraction of genomic DNA of maize and soybean, (2) amplification of the inserted genes, CaMV35S promoter and the NOS terminator inserted by means of the polymerase chain reaction (PCR) method, (3) amplification of the specific genes of maize and soybean in order to determine that the samples are maize and soybean, (4) characterization and confirmation of the PCR products by restriction enzyme analysis and the electrophoresis on agarose gel. The RR soybean contains CaMV35S promoter and NOS terminator, and the Bt-176 maize contains only CaMV35S promoter. Due to the high content of the starch in maize, the effect of the electrophororesis is not so good as of the soybean's. PMID- 12545758 TI - [Estrogenicity of trans-resveratrol in immature mice in vivo]. AB - To investigate the estrogenicity of trans-resveratrol in vivo, different doses of trans-resveratrol were administered orally (ig.) or subcutaneously (s.c.) to the weanling mice for 4 d. The results showed that 2.0 mg/kg.b.w of trans transveratrol (ig or s.c.) could shorten vaginal opening latency periods (P < 0.05) and enhance keratinization of vaginal epithelium dramatically, increase uterine wet weights and uterine-body weight ratios significantly (P < 0.05, P < 0.01). Additionally, trans-transveratrol could thicken the columnar epithelial cells or increase the numbers of glands in uterine in immature mice. It was concluded that trans-transveratrol could appear estrogenic actions in vivo. Moreover, its activity via subcutaneous administration was higher than that via oral administration. PMID- 12545759 TI - [Anti-oxidative effects of proanthocyanidins in mice induced by D-galactose]. AB - The anti-oxidative effect of proanthocyanidins(PC) was evaluated in D-galactose induced murine model. Male KUNMING mice were divided into different groups at random. The mice were induced by subcutaneous injection of D-galactose on the back of mice daily for 60 days and simultaneously PC and VE were administered to the mice by oral feeding. Results indicate that the lipid peroxide in blood, liver and brain significantly increased respectively because of the D-galactose injection, when compared to the control. The activity of SOD in RBC and liver significantly decreased while that of GSH-PX in blood and liver decreased. MAO-B in brain was significantly increased while MAO-B in liver doesn't change significantly. Oral feeding PC markedly decreased the formation of MDA in blood, liver and brain, increased the activities of SOD in blood and liver and decreased the activity of MAO-B in brain. Vitamin E also counteract the oxidative stress induced by D-galactose. In summary, treatment with PC at all the three tested doses were effective in exerting a protective effect against oxidative stress. PMID- 12545760 TI - [Study on the relationship between body index and blood pressure in Chinese adult]. AB - The authors use the anthropometry data of the Third National Nutrition Survey in China, and select the adult above 20-year-old as the study subjects in order to study the relationship between BMI and blood pressure value and to provide some reference data for defining the suitable BMI cut-off point of overweight and obesity in Chinese adult. The results show that the distribution of BMI varied with sex, age and areas; mean levels of systolic and diastolic blood pressure both gradually increased as BMI increased, multifactor linear regression analyses indicates there is obvious linear relationship between blood and BMI; The associations of BMI with high blood pressure were statistically significant after controlling for age, sex, area or smoking, logistic regression analyses indicate that odds ratios increased gradually at the different levels of BMI. These data quantify the strong associations of BMI with hypertension. PMID- 12545761 TI - [Studies on nano-particle sols of hydroxyaptite and titanium dioxide for haemo compatibility]. AB - A biological evaluation is conducted for two types of nano-particle sols, hydroxyaptite(HAP) and titanium dioxide(TiO2). The results show that HAP sol significnatly prolongs the bleeding time and coagulation time of mice as well as the prothrombin time(PT) and partial thromboplastin time(PTT) of rats while TiO2 sol exhibits no such effects. Neither HAP sol nor TiO2 sol instigated in-vitro hemolysis of rabbit erythrocyte. However, both of the materials caused in-vitro aggregation of rabbit erythrocytes. The reason underlying the different results as to the two types of material is their specific stabilizer, heparin for HAP sol and PVC for TiO2 sol. We came to the conclusion that a biologically inert stablizer has no less significance than the nano-particle's very own nature in a nano-material's application prospect. PMID- 12545762 TI - [Changes of both knowledge levels and health-related behaviors of preventing cardio-cerebro-vascular diseases in population after intervention]. AB - Comprehensive intervention measures for preventing cardio-cerebro-vascular diseases of community population were carried out in Beijing, Shanghai and Changsha urban areas, and then was evaluated to its effects on knowledge, attitude and behavior (KAB) levels and health-related behaviors of population by quasi-trial designation. Results showed that the net KAB leveling were separately 9.89 in 1995, 4.74 in 1997 and 10.66 in 2000 in comparison with base investigation, and statistically significant difference was found. Of 6 health related behaviors, caring to health knowledge, being influenced by health knowledge and exercising were significantly influenced, next controlling salt consumption. No change of smoking and alcohol drinking was found. Among different target population, these changes among women were higher than man as well as at home or retired than at work, and hypertension than normtension. As aging and education time decreasing, both absolute and relative effects of health education were better. It suggests that comprehensive intervention with health education as its main measure promote KAB levels and transfer health-related behaviors of population. It is great important to preventing cardio-cerebro-vascular diseases of community population. PMID- 12545763 TI - [Study on solid phase extraction spectrophotometric determination of copper(II) with 2-(4-antipyrinylazo)-5-dimethylaminoaniline]. AB - Based on the color reaction of 2-(4-Antipyrinylazo)-5-Dimethylaminoaniline (ADA) with copper (II) and the solid phase extraction of its colored complex with Waters Porapak(r) Sep-Park-C18 cartridge, a new method for the determination trace amount of copper(II) was studied. In the medium of Tween-80 and presence of pH = 4.5 buffer solution, ADA can react with copper (II) to form a stable 2:1 complex. The colored complex can extracted by Porapak(r) Sep-Park-C18 cartridge and eluted by ethanol contain 1% of acetic acid, then can be determined by spectrophotometer at 530 nm. Beer's law is obeyed in range of 0.05(1.0(g/ml. This method can be applied to the determination trace amount of copper in water with satisfactory results. PMID- 12545764 TI - [Effects of L yaojiu of invigorating blood circulation and eliminating blood stasis in old rats]. AB - L Yaojiu was investigated for its effects of invigorating blood circulation and eliminating blood stasis using old rats (aged over ten months) as the model of blood stasis. Three different dosages of L Yao Jiu (14.3, 28.5, 57.0 mg crude drugs/kg) had been administrated orally per day to rats for 12 consecutive days. The changes of the mesenteric microcirculation and the hemorrheology were observed. The content of serum malondialdehyde (MDA) and the activities of serum monoamine oxidase (MAO) and superoxide dismutase (SOD) were determined. The results showed L Yao Jiu could accelerate the blood streams, dilate the capillaries and inhibit erythrocyte aggregations. It also reduced blood viscosity, inhibited platelet aggregations and improved the function of erythrocytes. On the other hand, the activities of total-SOD and Cu-SOD were increased and the content of MDA was decreased. These results suggests that L Yao Jiu could invigorate blood circulation and eliminate blood stasis in old rats. PMID- 12545765 TI - [Effect of alcohol extract of Zingben officinale rose on immunologic function of mice with tumor]. AB - In order to study the effect of alcohol extract of Zingben officinzle rose on immunologic function of mice with tumor, mice are orally administrated with dose of 10 g/kg and 40 g/kg alcohol extract of Zingben officinale rose. The thymus index, spleen index, percentage of phagocytosis, rate of alpha-ANAE+ and level of hymolysin(IgM) are use in the study as the parameters to evaluate the effect of alcohol extract of Zingben officinale rose on non-specific and specific immune function of mice with tumor. The results indicate that alcohol extract of Zingben officinale rose can raise significantly the thymus index, spleen index, percentage of phagocytosis, rate of alpha-ANAE+ and titer of IgM of mice with tumor indicating that alcohol extract of Zingben officinale rose can improve significantly the status immunologic function in mice inadequacy of mice with tumor. PMID- 12545766 TI - [Study on the spectrophotometric determination of microamounts of iron with 1-(2 pyridylazo)-2-naphthol-6-sulphonic acid]. AB - A new spectrophotometric method has been developed for the highly selective determination of Fe2+ with 1-(2-pyridylazo)-2-naphthol-6-sulphonic Acid. At pH 4.0, Fe2+ an be selectively determined at the wavelength of 760 nm. The calibration curve is liner in a concentration range of 0-10 micrograms/10 ml, relative coefficient is 0.9997, the apparent molar absorptivity is 1.57 x 104 L.mol-1.cm-1, RSD of within-run and between-day were 1.66% and 2.51%, the average rate of recovery 101.1%. The method has been applied to the direct determination of iron in milk powder, common coexistent substances don't interfere with the determination. PMID- 12545767 TI - [Biological synthesis and application of bacteriocins]. AB - Bacteriocins produced by lactic acid bacteria are a heterogeneous group of peptide inhibitors. Most of bacteriocins have relatively narrow inhibitory spectra. A few of them have wider inhibitory spectra, being active against a wide range of gram-positive bacteria. Bacteriocins can be divided into three types. There are small heat-stable peptides, large heat-labile proteins, and lantibiotics. Their physico-chemical nature, genetic behavior and application are reviewed. PMID- 12545768 TI - [Toxicity of microcystin and its bioaccumulation effect]. AB - Microcystins are cyclic heptapeptide hepatotoxins from certain cyanobacteria. It can inhibit hepatic protein phosphatase 1 and 2A activities, break the balance between protein phosphorylation and phosphorylysis, causing liver damage even necrocytosis. Furthermore, it is a strong tumor promoter. Yet, its hazard received little study. This paper presents a latest review of toxicity and bioaccumulation of Microcystins. PMID- 12545769 TI - [Conformational flexibility of enzyme active sites]. AB - Comparative studies of conformation and activity changes of a number of enzymes during denaturation have shown that inactivation generally precedes detectable global conformational changes. Kinetically, the inactivation rates of enzymes during denaturation are much faster than the rates of global conformational changes under identical conditions. It is suggested that the conformation of enzyme active sites are held together by weaker forces and consequently more flexible compared to the molecule as a whole. Conformational changes at the active sites were demonstrated directly by fluorescent and spin probes introduced at the active site of creatine kinase, glyceraldehyde-3-phosphate dehydrogenase and ribonuclease A. In addition, the susceptibility of ribonuclease A to proteolysis is markedly increased in dilute GuHCl. By separation and N-terminal sequence analysis of the peptide fragments liberated by hydrolysis with trypsin or proteinase K, the cleavage points can be identified to show that without gross unfolding of the RNase molecule as a whole, loosening up of active site conformation has indeed occurred during inactivation in dilute GuHCl. For the role of active site flexibility in enzyme catalysis, it is possible that each intermediate step of the whole cycle of catalysis requires the enzyme molecule to be in a different conformation state. Active site flexibility would therefore be essential for the full expression of enzyme activity. It has recently been demonstrated that conformational change, especially that at the active site, accompanies enzyme catalysis and the activation of a number of enzymes involves the loosening up of the active site structure. PMID- 12545770 TI - [Peripheral effector mechanism hypothesis on cardiovascular dysfunction after spaceflight]. AB - In the years of 1990's, we systematically studied the adaptational changes in structure and function of both the heart and the vessels during simulated weightlessness. In our serial work, the tail-suspension rat model was used to simulate the microgravity-induced cephalad shift and redistribution of blood. On the basis of the facts we observed and the more recent advances in space and ground-based studies in 1990's, we put forward a hypothesis to offer a possible explanation for the frequent occurrence of postflight cardiovascular dysfunction. It states that, in addition to the factor of hypovolemia, the microgravity induced adaptational changes in the structure and function of the two main effectors of the cardiovascular system, i.e., the arterial smooth muscle and the cardiac muscle might be one of the most important mechanisms accounting for postflight cardiovascular dysfunction. PMID- 12545771 TI - [TNF-related apoptosis inducing ligand and its research progress on cancer treatment]. AB - TRAIL (TNF-related apoptosis inducing ligand) belongs to the tumor necrosis factor (TNF) cytokine family, can induce apoptosis in a wide variety of tumor cell lines. This review introduces research progress on TRAIL from several aspects: the structure and function of TRAIL, the pathway of its inducing apoptosis, TRAIL and cancer treatment, and its foreground. PMID- 12545772 TI - [Research progress in gene engineered antibodies]. AB - Gene engineered antibodies(GEAs) have been applied in a lot of aspects with the great development of the technique of GEAs. The technique of GEAs such as ribosome display and phage surface display and their application progress were summarized in this review. PMID- 12545773 TI - [Effects of estrogen in the central nervous system]. AB - Estrogen exerts a variety of effects (including electrophysiological, neutrophic and metabolic effects) on neurons in the adult central nervous system. With the many progresses made in the study of estrogen signaling through both genomic and nongenomic pathways, awareness of estrogen's effects on neural function is broadening rapidly. And it also helped people to understand more about the mechanisms by which estrogen regulating GnRH neurons, inducing or maintaining dendritic spine in hippocampus and protecting neurons from a variety of insults. Recently, epidemiological studies have suggested that estrogen, as a hormone replacement therapy in postmenopausal women, may protect against Alzheimer's disease (AD). Findings in these areas demonstrate a role for estrogen that is beyond direct control of reproductive function. PMID- 12545774 TI - [Augmenter of liver regeneration: a novel cytokine]. AB - Augmenter of liver regeneration (ALR) is a novel cytokine which specifically stimulates hepatic cell proliferation and is able to rescue acute liver failure caused by carbon tetrachloride. This article reviews the discovery, tissue distribution, functions and prospect of ALR and its relationship with hepatic stimulatory substance (HSS). PMID- 12545775 TI - [Calcium dependent synaptic plasticity]. AB - Recent work shows that the intracellular free Ca2+ concentrations ([Ca2+]i) of the presynaptic and postsynaptic neurons play crucial signaling roles in short- and long-term synaptic plasticity. Residual [Ca2+]i followed conditioning stimulation may cause short-term synaptic enhancement. Presynaptic [Ca2+]i could influence the replacing of presynaptic depressed vesicles, as well as encode the precise relative timing of presynaptic input and postsynaptic activity and generate long-term synaptic modification of opposite polarity(LTP or LTD). PMID- 12545776 TI - [Progress in the study of obesity]. AB - Obesity has been rising as a common social phenomenon. Recent research has convinced us that rather than a cosmetic problem, obesity should be considered as a real disease which is a complex disorder of appetite regulation and energy metabolism. Actually, obesity is caused by many factors such as heredity, environment and food contents, among which hereditary gene seems to work as a key element. Induced and followed by complicated factors, obesity is particularly associated with diabetes mellitus II, coronary heart disease, certain forms of cancer and so on. At present, the access to treat obesity is still in its early stage and the function of leptin identified in 1994 is also the focus of debate. This review provides a comprehensive insight into the epidemics, etiology, healthy problems, treatment strategies of obesity, and leptin as well, attempting to develop an outline of obesity research. PMID- 12545777 TI - [Obesity and the central nervous system regulation]. AB - There are increasing evidences to show that central nervous system is involved in the regulation of energy homeostasis. Energy intake is usually matched to energy expenditure over a period of time. Obesity occurs when the amount of energy intake (or food intake) is more than the energy expenditure. Because of the enormous tolls on human health taken by obesity and related disorders, an improved understanding of the control of food intake is an important priority. The aim of this article is to briefly review the advances in recent years on long term maintenance of energy homeostasis and the role of central nervous system. In the present review, the following contents are included: (1) satiety and its production, (2) adiposity signals and the regulation of food intake, (3) nuclei in central nervous system involved in food intake, (4) the first- and the second order neuronal signaling in hypothalamus on control of food intake and (5) clinic implications. PMID- 12545778 TI - [Role and regulation of calcineurin-dependent signal pathway in cardiac hypertrophy of rats]. AB - The present study investigated the role and regulation of calcineurin-dependent signal pathway in cardiac hypertrophy of rats from the three levels of animal model, culturing cells and molecular biology. The results showed as follows: (1) Calcineurin signal pathway involves in myocyte hypertrophy induced by various factors such as hemodynamic overload, myocardial fibrosis, paracrine/autocrine factors, etc.; (2) Calcineurin-dependent signal pathway plays an important role not only in AngII- and bFGF-induced cardiac myocyte hypertrophy but also in AngII and bFGF-stimulated cardiac fibroblast proliferation. (3) Calcineurin pathway may associate with MAPK and PKC pathways at several levels; (4) Activation of calcineurin depends on sustained increase of intracellular calcium concentration and is regulated by protein phosphorylation. The expression of calcineurin gene in AngII-stimulated myocytes might be regulated by Ca2+ signal and MAPK cascade. In conclusion, Ca(2+)-calcineurin signal pathway involves in the development of cardiac hypertrophy of rats. PMID- 12545779 TI - [Advances in the study on the proteins related desensitization and internalization of G-protein coupled receptor]. PMID- 12545780 TI - [Biological effects of urotensin II]. PMID- 12545781 TI - [The structure, function and activity regulation of hypoxia-inducible factor-I and its relationship with hypoxia signal transduction]. PMID- 12545782 TI - [Hypoxic response pathway]. PMID- 12545783 TI - [Progress of study in estrogen receptor-beta]. PMID- 12545784 TI - [Chemical modification of hemoglobin and blood substitutes]. PMID- 12545785 TI - [Interstitial cells of Cajal and gastrointestinal motility generation]. PMID- 12545786 TI - [The role of N-ethylmaleimide-sensitive fusion protein (NSF) in cell secretion]. PMID- 12545787 TI - [Progresses in IkB kinase and related signal transduction]. PMID- 12545788 TI - [Current status of the study of ischemic preconditioning]. PMID- 12545789 TI - [Neuregulin/ErbB signal transduction pathway in the development of nervous system]. AB - Neuregulins are a family of structurally-related polypeptides encoded by four distinct genes. Three isoforms of neuregulin (NRG1, 2, and 3) are highly expressed in the developing and adult brain, among which NRG1 is best characterized. NRG1 has multiple functions essential for the development of the nervous system. It promotes growth and differentiation of glial cells and regulates the migration of cerebellar granule cells along radial glial fibers. During synaptogenesis, NRG1 induces the expression of acetylcholine receptor at the neuromuscular junction and in CNS, and expression of the NMDA receptor NR2C subunit and GABAA receptors in cerebellar granule cells. Recent studies indicate that NRG receptors are densely distributed in the postsynaptic membrane, suggesting that NRGs may play an important role in synaptic plasticity. We review recent progress in studies of NRG, their functions, and signaling mechanisms. PMID- 12545790 TI - [Zinc ions: an endogenous neuromodulator]. AB - Zinc ions are widely distributed in the central nervous system. Zinc ions are released from neurons in a calcium-dependent manner. In recent years abundant evidence indicates that zinc ions modulate not only the release of transmitters, but also the voltage- and ligand (excitatory and inhibitory amino acids)-gated channels, suggesting that these ions may serve as an important endogenous neuromodulator. PMID- 12545791 TI - [Progress in the study of pain by brain imaging]. AB - This review deals with the recent functional brain imaging studies of pain. In summary, the sensory-discriminative component of pain is related with the lateral thalamus, primary and secondary somatosensory area and insular cortices, while the posterior parietal and prefrontal cortices seem to play a role in the cognitive-attentional process of nociceptive information. Different parts of anterior cingulate gyrus are correlated with cognitive and emotional aspects of pain. Brain imaging data obtained from clinical patients suffering from various kinds of pain especially neuropathic pain, were discussed at the end of this article. PMID- 12545792 TI - [Strategies and methods to identify genes for complex diseases]. AB - Genes underlie numerous human diseases and traits. Although we have witnessed a great deal of success in identifying disease-susceptible genes, the task remains challenging for most of the complex diseases. This paper reviews evidence for the role of genetic factors in complex diseases, and strategies that can potentially optimize our chance of success in identifying genes involved in complex diseases. Advances in molecular biology, particularly mapping of the human genome, statistical methods that provide more accurate models of complex patterns of inheritance, and advances in basic medical science, which have increased our understanding of disease pathophysiology, will ultimately strengthen the ability of the current generation of genetic epidemiological studies to identify the genetic basis of complex human disorders. PMID- 12545793 TI - [Molecular mechanisms involved in the regulation of telomerase activity]. AB - The main components of telomerase include RNA subunit, hTERT catalytic subunit and hTEP1 regulatory protein. While telomerase plays an important role in maintenance of telomere structural stability, telomere is also involved in the regulation of telomerase activity. Some anticancer drugs inhibit the telomerase activity by breaking the structure of telomere. Activation of human telomerase requires de novo transcription of hTERT gene and correct incorporation of all subunits into holoenzyme. The expression of telomerase activity is likely to be regulated at multiple levels. Firstly, the expression and transcription of the hTERT gene is regulated by various factors; Secondly, various oncogenes and tumor suppressor genes are involved in regulation of telomerase activity by protein protein interaction with telomere and telomerase proteins directly or indirectly. Thirdly, phosphorylation of hTERT and hTEP1 by protein kinase leads to significant up-regulation of telomerase activity, and conversely, the dephosphorylation result in down-regulation of telomerase activity. PMID- 12545794 TI - [Analgesic mechanisms of noradrenaline in the spinal dorsal horn]. AB - The superficial laminae of the spinal dorsal horn play important roles in nociceptive transmission and modulation. Noradrenergic fibers originated from brainstem pain-control system terminate and release noradrenaline (NA) in the superficial dorsal horn. The released NA might take part in the modulation of nociceptive transmission through the following pathways: (1) inhibits the glutamate and substance P release from primary afferent terminals; (2) increases the release of inhibitory neurotransmitters from lamina II (substantia gelatinosa) neurons. Morphological findings also raise the possibility that NA directly inhibits projection neurons in laminae I/III which convey the nociceptive information to the thalamus, thus inhibit nociceptive transmission at spinal level. PMID- 12545795 TI - [New type of two-hybrid systems in protein-protein interaction studies]. AB - Some new type of two-hybrid systems, such as split-ubiquitin system, protein fragment complementation assay, repressor reconstitution assay and SOS recruitment system, have been developed recently. Similar to the original transcription-based yeast two-hybrid system, these systems are to establish an assay for protein-protein interactions, in which some functional proteins can be split into two parts in structure and their activities can be recovered by reconstitution. Due to their non-transcriptional properties, these new types of two-hybrid systems have become useful extension of the yeast two-hybrid system and powerful tools for the study of protein interactions. PMID- 12545796 TI - [The role of immune cells and inflammatory mediators in pathogenesis of intestinal inflammation]. AB - The initiators of intestinal inflammation are greatly variable, but the mechanisms underlying the immunologically mediated mucosal damage are similar. A great progress in our understanding of the functions of gut-associated lymphoid tissue was achieved because of the advances in immuno-biology and molecular immunology. At the beginning of an inflammation microorganisms and hither to derived products or allergen firstly stimulate and activate the specific and nonspecific immune cells, and then intestine epithelials, macrophages and lymphocytes release various cytokines. These cytokines are able to recruit more immune cells to be activated and more cytokines to be released. A "cascade" is then generated between epithelials, macrophages and lymphocytes. Finally the inflammation in certain area of intestinal tract is generated. The balance between inflammatory factors and anti-inflammatory molecules depends the results of the inflammation. Therefore, the better understanding of the mechanisms on inflammatory and anti-inflammatory factors is helpful for the diagnosis and treatment of intestinal inflammation. PMID- 12545797 TI - [Synaptic transmission in retina bipolar cell]. PMID- 12545798 TI - [Microphysiometer--a real-time, sensitive method for evaluation of the functional activity of cells]. PMID- 12545799 TI - [Interference with the formation of the epididymal microenvironment--a new strategy for male contraception]. PMID- 12545800 TI - [Progress in study of human immortalized cell repository]. PMID- 12545802 TI - [Signal transduction of insulin receptor]. PMID- 12545801 TI - [Receptor activity modifying protein of calcitonin gene-related peptide family]. PMID- 12545803 TI - [C-type natriuretic peptide and cardiovascular disease]. PMID- 12545804 TI - [Recent progresses on the modulation of estrogen to dopaminergic neurons]. PMID- 12545805 TI - [Recent advances in cardiac beta 2-adrenergic receptor]. PMID- 12545806 TI - [Opioid tolerance: a phenomenon of latent hyperalgesia]. PMID- 12545807 TI - [Experiences in scientific research]. PMID- 12545808 TI - [The modulation of type I collagen and its receptor system on biological characteristics of osteoblasts]. AB - This paper addresses the question whether type I collagen-receptor system is necessary for the functional activity of osteoblasts. Through blocking type I collagen-receptor system by type I collagen antibody or integrin alpha 2 beta 1 antibody, the proliferation of osteoblasts was studied by cell count, the apoptosis was studied by flow cytometry, and the mRNA expressions of type I collagen, integrin alpha 2 beta 1 and osteocalcin were also detected by RT-PCR techniques. When type I collagen-receptor system was blocked, osteoblasts showed high apoptosis rate, lower growth kinetics and weak mRNA expression of type I collagen, integrin alpha 2 beta 1 and osteocalcin; the blocking effect was reversible. It suggests that type I collagen-receptor system is necessary for osteoblastic phenotype. The biomaterials for bone tissue engineering should be constructed according to the extracellular matrix of osteoblasts, which supply normal extracellular environment for osteoblasts. PMID- 12545809 TI - [Enhancement of BCG-induced hBD-1 mRNA expression in human pulmonary gland epithelial cells]. AB - Using reverse-transcription PCR (RT-PCR) and Northern blot analysis, the present authors found an enhanced expression of the human beta-defensin 1 (hBD-1) gene in the SPC-A-1 cells challenged with heat-killed M. Bovis Bacille Calmette-Guerin (BCG). The enhanced expression of hBD-1 mRNA was dose- and time-dependent. This result indicated a significant progress in the work of developing a new antimicrobial strategy that enhances mucosa antibiotic peptide expression for the prevention and treatment of mucosal infections. PMID- 12545811 TI - [Preparation of 131I-VIP and 131I-VIP receptor imaging]. AB - This study was aimed at the preparation of 131I-vasoactive intestinal peptide (VIP) and its preliminary application in clinical imaging. VIP was labeled with Na 131I using chloramine-T method, then isolated by Sephadex G-10 column chromatography and examined by silica 60F254 thin layer chromatography. The bacteria and pyrogen were examined and the safety test was carried out. One control and two patients suffering from abdomen tumor were investigated. The results showed that the labeling rate of 131I was 80% and the specific activity of 131I-VIP was 36 TBq/mmol. The radiochemical purity of 131I-VIP was over 98%, and it decreased to 95% after six hours' storage at 4 degrees C. It was proved that the 131I-VIP eluate had no bacteria, no pyrogen and no poison. The injected 131I-VIP was distributed into the lungs immediately and was eliminated through kidneys. The primary tumor could be visualized about half an hour to 3 hours after injection. This study demonstrates that 131I-VIP is suitable for in vivo imaging and may be used as an effective tracer to identify the tumor site in patients with VIP receptor positive carcinoma. PMID- 12545810 TI - [Dynamic study of hepatocellular pathological change and uptake rate of 125I insulin during experimental hepatocarcinogenesis]. AB - This study was designed to observe the hepatocellular pathological change and uptake rate of 125I-insulin during experimental hepatocarcinogenesis in rats and clarify the possible mechanism of increasing uptake of 125I-insulin. 80 SD rats were divided into 2 groups, control and experimental groups. All of the rats were given common feed, and the rats of experimental group were given extra diethylnitrosamine (DENA) 70 mg/kg each week. At 6th, 11th, 15th and 20th week after start of the experiment, 10 control and 10 experimental rats were killed 1 hour post administration of 125I-insulin by tail vein. Blood, liver, lung, spleen, kidney, muscle and bone were collected; the radioactivity was measured and calculated %ID/g. The data of 2 groups were compared and examined with t test. All of livers were pathologically examined. The results showed all livers of control group were normal. At 6th week, the surfaces of experimental rats' livers were coarse. Hepatocellular hyperplasia was observed, 125I-insulin-uptake rate was 1.86 time as much as that of control group. At 11th week, the liver's colour became lighter than that of control group. Hyperplasia and hepatocirrhosis were observed, 125I-insulin-uptake rate was 1.50 time as much as that of control group. At 15th week, hyperplastic nodules were observed in all experimental rats' livers. Hepatomacellulae were observed in 6 rat livers. 125I-insulin-uptake rate was 1.56 time as much as that of control group. At 20th week, the livers became enlarged out of shape. There were a lot of big or small greyish white nodules in all livers. Necrosis, liquefaction and hemorrhage were observed. Hepatomacellulae were observed in all of experimental rats livers. 125I-insulin-uptake rate was 1.46 time as much as that of control group. The differences of 125I-insulin uptake rate between experimental and control groups were significant. These results demonstrated that the liver ability of uptaking 125I-insulin increased which mainly took place during the period of hepatocellular hyperplasia, and the hepatomacellulae kept this characteristic. PMID- 12545812 TI - [Experimental studies on exons 5-8 of p53 gene mutation in laryngeal squamous cell carcinoma]. AB - This study was designed to detect the point mutations of exons 5-8 of p53 gene in laryngeal squamous cell carcinoma (LSCC) and analyze their relationship. The detection of fresh tumor samples from LSCC patients was performed using silver staining PCR-SSCP method. From among 60 patients samples, 47 were positive in SSCP. Mutation rate was 78.3% (47/60). The results showed that the prevalence of p53 mutations in LSCC subjected to silver staining PCR-SSCP test were 50% (30/60) in exon 5, 11.67%(7/60) in exon 6, 41.6%(25/60) in exon 7, and 25%(15/60) in exon 8. The majority of the mutations were found in exon 5 and exon 7. Exon 5 and exon 7 of p53 gene may be the mutation hotspot in LSCC; they may be the critical position easily attacked by some carcinogen factors relating to LSCC. PMID- 12545813 TI - [Relationship of p53 gene mutation with pathological characteristics and prognosis of thyroid carcinoma]. AB - This study was directed to the role of p53 gene in the carcinogenesis of thyroid carcinomas and to the correlation between p53 gene and the clinicopathological characteristics of the cancer. Single-stranded conformation polymorphism of PCR was used in detecting p53 gene point mutations in exons 7, 8. The result showed that ten of thirty-one thyroid carcinomas had mutations in exons 7 and 8 (32.3%). The frequency of p53 gene mutations was significantly higher in relapse group than in no relapse group (P < 0.01). No statistically significant differences in p53 mutation were found relating to metastasis, histological type and differentiation (P > 0.05). These data suggest that the mutation of p53 gene may play an important role in thyroid carcinoma and the mutations of p53 gene be associated with the prognosis of thyroid carcinoma. PMID- 12545814 TI - [Studies on thymidine kinase gene (TK) and GCV system for treatment of human retinoblastoma (RB)]. AB - The antitumor effect of herpes simplex virus thymidine kinase (HSV-TK)/GCV system on human retinoblastoma (RB) was studied here. A retroviral vector with tk gene (pLXSN-TK) was transduced into packaging cell line PA317. Recombinant retroviral was obtained and employed to infect human RB cells. The in vitro efficacy of TK/GCV was evaluated by survival rate of RB cells with and without TK transduced 5 days after treated with GCV. A nude mouse model with heteroplantation of human RB was established to examine the in vivo efficacy. Mice with RB were given an in situ injection of retrovirus followed by treatment with GCV for 14 days (50 mg/kg). The RB/TK cells in tissue culture dish showed far more sensitive to GCV than RB cells. The tumors in RB mice with TK gene transduced were much smaller than those in control. The results indicate that HSV-TK/GCV system can suppress growth of RB both in vitro and in vivo. It could be a valuable method for treatment of RB patients. PMID- 12545815 TI - [Expression and implication of c-myb and bcl-2 proteins in C6 gliomas]. AB - This study was aimed at the level and implication of expression of c-myb and bcl 2 proteins in C6 glioma. The therapeutic effect of oncogene c-myb antisense oligodeoxynucleotide on C6 glioma in nude mice was studied. C6 glioma cells were implanted into the cutis of 36 nude mice. Ten days later, the nude mice were randomly divided into three groups and were treated with SON, AON and normal saline respectively. Three mice of every group were killed at 4 days, 8 days, 12 days and 16 days after treatment, respectively. The levels of c-myb and Bcl-2 proteins expression in the three groups were observed by the S-P immunohistochemical method. The results showed the expression of c-myb and bcl-2 proteins was significantly decreased in the AON group, compared with that in the SON and control groups (P < 0.05). These data suggest that c-myb and bcl-2 proteins may play an important role in malignant glioma, and c-myb gene and bcl-2 gene may be involved in the tumorigenesis and development of glioma. PMID- 12545816 TI - [A study on the inactivation of p16 genes and the expression of P16 protein in primary hepatocellular carcinomas]. AB - The aim of the present study was to determine whether p16 gene is involved in the genesis of primary hepatocellular carcinoma (HCC). Twenty-five primary HCC tumor and corresponding non-tumor liver tissue specimens were examined for P16 gene alterations. The identification of deletion of p16 gene exon 1 and exon 2 was performed using comparative multiplex polymerase chain reaction (PCR) analysis. The point mutation of p16 gene exon 2 was investigated by single strand conformational polymorphism(SSCP) analysis, and the status of p16 gene methylation was screened using PCR-based methylation analysis. Moreover, 35 parafin specimens of primary HCCs with corresponding non-tumor liver tissues, including the 25 cases described above for screening of p16 gene alterations, were investigated also for P16 protein expression by using immunohistochemical analysis. The results of comparative multiplex PCR analysis showed that 12% (3/25) p16 gene deletions, including homozygous deletions in 2 tumors and hemizygous deletion in 1 tumor, were found in the primary HCCs samples. No point mutation was identified in the remaining 22 tumor samples without p16 gene deletions by using SSCP analysis. Hypermethylation was detected in 24% (6/25) of tumor samples by PCR-based analysis. However the corresponding non-tumor liver tissue specimens were always unmethylated at p16 locus. Loss of P16 protein expression, detected by immunohistochemistry, occurred in 16 of 35 (45.7%) tumor samples, whereas all the non-tumor liver tissue specimens showed positive p16 staining. These results indicate that inactivation of p16 gene is relevant to the genesis of HCC. PMID- 12545817 TI - [Determination of malignant disease-associated DNA-binding protein 2 in patients with liver cancer]. AB - The aim of this study was to assess the value of malignant disease-associated DNA binding protein 2(MAD2) in the diagnosis of liver cancer. The concentration of plasma MAD2 was determined in 27 patients with primary liver cancer, 14 patients with metastatic liver cancer and 12 healthy subjects by ELISA assay. The primary tumors of all the patients with metastatic liver cancer were located in the gastrointestinal tract, which had been radically resected and had no signs of local recurrence. The concentrations of plasma MAD2 in the patients with primary liver cancer, with metastatic liver cancer, and the healthy subjects were 30.56 +/- 11.38 micrograms/ml, 9.27 +/- 5.58 micrograms/ml and 8.43 +/- 5.62 micrograms/ml, respectively. The concentration of MAD2 in patients with primary liver cancer was significantly higher than that in the patients with metastatic liver cancer and healthy subjects (P < 0.001). There was no significant difference between the healthy subjects and the patients with metastatic liver cancer(P > 0.05). These results suggest that MAD2 may be an useful marker for the diagnosis of primary liver cancer. PMID- 12545818 TI - [Effects of human chorionic gonadotropin on cellular cAMP levels in human gastro adenocarcinoma SGC-7901 cells]. AB - The purpose of this study was to determine the effects of hCG on cellular cAMP levels in gastric adenocarcinoma cells(SGC-7901). Radioimmunoassay was used. The results showed that hCG obviously increased cAMP levels in SGC-7901 cells in a dose-dependent and time-dependent manner. When the concentration of hCG increased from 10(-11) mol/L to 10(-6) mol/L, the cellular cAMP levels increased from about 2.1 pmol/mg protein to about 10 pmol/mg protein and were significantly different from the control(P < 0.01). These results suggest that hCG possibly exerts its effects on gastric cancer cells by activating adenylyl cyclase/cAMP signal transduction pathway. PMID- 12545819 TI - [Transcription regulation of intercellular adhesion molecule (ICAM-1) on lymphatic endothelial cells of rectum adenocarcinoma of human]. AB - This study inquired into the mechanism of cancer lymphatic metastasis. The immunohistochemistry method and DNA hybridization in situ were employed in examining the peritumoral rectum tissues and metastatic lymph nodes from 8 rectum carcinoma patients and the rectum tissues and lymph nodes from 5 normal subjects. The results showed that the proteins of ICAM-1 and nuclear factor kappa B (NF kappa Bp65) were expressed on the lymphatic endothelial cells of the rectum adenocarcinoma patients, and there was a NF kappa B binding consensus sequences on ICAM-1 promoter in lymphatic endothelial cells of rectum adenocarcinoma patients. When DIG-AKP labeled 38 bp oligonucleotide probe was used, there was no the expression of ICAM-1 and NF kappa Bp65 on the lymphatic endothelial cells of the normal human lymph node and rectum tissues. It is suggested that the activation of ICAM-1 promoter may critically depend on NF kappa Bp65 homodimers or heterodimers binding to a variant kappa B site on the lymphatic endothelial cells of the rectum adencarcinoma of human. These observations may indicate the potential roles of NF kappa B in cancer metastasis, thus giving clues to facilitate the development of a novel anti-metastasis strategy, that is obstruction the NF kappa B-activation pathways. PMID- 12545820 TI - [Expression of c-fos mRNA following moderate lateral fluid percussion brain injury in rats]. AB - This experiment was designed to study the expression of c-fos mRNA in brain following moderate lateral fluid percussion brain injury in rats and to observe the temporal pattern of its expressions following percussion. Male Sprague-Dawley rats were divided into normal control, sham operation control and injury groups. The rats of the injury group were subjected to moderate lateral fluid percussion injury (0.2 MPa). The injury group was then subdivided into 5 min, 15 min, 30 min, 1 h, 2 h groups according to the time elapsed after injury. The expression of c-fos mRNA was studied with reverse transcription polymerase chain reaction(RT PCR) semi-quantitatively. c-fos mRNA in cortex and brain stem was expressed weakly in control groups. After 5 min of percussion, the expression of c-fos mRNA increased progressively and remained elevated up to 2 h after brain injury. This result suggested that the induction and expression of the c-fos mRNA in cortex and brain stem after fluid percussion brain injury were increased rapidly. The temporal pattern of induction in cortex was similar to that in brain stem. PMID- 12545821 TI - [Distribution of peptidergic neurons in the rat suprachiasmatic nucleus]. AB - The distribution of the peptidergic neurons in the suprachiasmatic nucleus(SCN) in five adult male SD rats was examined by multi-immunolabelling for vasoactive intestinal polypeptide(VIP), arginine vasopressin (AVP) and somatostatin(SOM). The immunoreactions were visualized with an Elite ABC Kit(Vector Lab). The substrates in the peroxidase reaction were diaminobenzidine(DAB), tetramethylbenzidine(TMB) and 4-chloro-1-naphtol. The results showed that the differences in distribution and population of VIP-, AVP- and SOM-like neurons existed in the rostral, intermediate and caudal regions of the SCN and these neurons were overlapped and SOM-like neurons were scattered among them in the area between VIP- and AVP-like neurons. These findings may provide new morphologic evidences for explaining the functional mechanism in the SCN. PMID- 12545822 TI - [The alterations in vasoactive intestinal polypeptide-immune reaction positive neurons of cerebral tissues in epileptic rats induced by penicillin]. AB - To further explore the action of vasoactive intestinal polypeptide(VIP) in epileptogenesis, we made an immunocytochemical analysis and observed the alterations in VIP-energic neurons of the cerebral cortex, hippocampus and amygdaloid in rats. The animals were divided into three groups: the control, the epileptic group in which seizure was induced by injected Penicillin(PNC) intraperitoneally, and the nimodipline(NIM) group in which seizure was impressed by giving PNC after NIM, a dihydropyridine calcium entry blocker, was injected. The results showed that the number of neurons of epileptic group increased, compared with that of control group (P < 0.01), the neuronic number of the epileptic group was higher than that of the NIM group(P < 0.01), whereas there was no significant difference in neuronic number between the control group and the NIM group(P > 0.05). These suggest that VIP and Ca2+ participate in the process of epileptogenesis. PMID- 12545823 TI - [The expression of trkC in spared dorsal root ganglion following unilateral dorsal root rhizotomy]. AB - This study sought to explore the expression of trkC in spared dorsal root ganglion(DRG) following unilateral dorsal root rhizotomy. 15 cats were divided into three groups: the intact group, the 3-day and 10-day groups after operation (unilaterral L1-L5 DRG and L7-S2 DRG were sectioned, but L6 DRG was spared). The animals were sacrificed on the 3rd day and 10th day after operation, respectively. The L6 DRG from the three groups were taken and cut into frozen sections (20 microns). The sections were stained using specific antiserum for trkC by the immunohistochemical ABC method. The numbers of large sized neurons (> 57 microns) and small sized neurons (< 42 microns) for trkC-IR were counted respectively. The results showed that the number of positive large sized neurons for trkC of the 10-day group after operation decreased significantly than that of the intact group and the 3-day group after operation, while the number of positive small sized neurons for trkC of the 10-day group increased apparently than that of the intact group and the 3-day group after operation (P < 0.05). This experiment demonstrated that unilateral partial dorsal root rhizotomy could change the expression of trkC might in large neurons and small sized neurons, and the results suggested that trkC might involve in the spinal cord plasticity following partial dorsal root rhizotomy. PMID- 12545824 TI - [An evaluation of adenosine triphosphate bioluminescence assay for tumor in vitro chemosensitivity testing]. AB - This study was aimed at the feasibility of using ATP-bioluminescence assay for tumor in vitro chemosensitivity testing. With the use of this assay, the authors determined dose-response curve in mouse fibroma cell line L929 treated with chemotherapeutic agents, and investigated the different in vitro responses of 6 ovarian carcinomas (5 from fresh tumor tissues, 1 from ascites) treated with etopside, cis-plating, 5-fluorouracil and adriamycin. The results showed that the coefficients of variation for triplicate assays ranged from 1.2% to 15.8% which means high reproducibility of the assay. The single cell suspension (including < 30 cells clusters) could be separated from tissue fragments by means of enzyme cocktail (collagenase, Dnase, pronase). The viable cells were over 90%. This study demonstrates that ATP-bioluminescence assay is a sensitive, reliable and efficient method for tumor chemosensitivity testing. In this connection, the correlation between in vitro drug sensitivity and in vivo patient response is worth further studying. PMID- 12545825 TI - [A clinical study on PA-MSHA vaccine used for adjuvant therapy of lymphoma and lung cancer]. AB - The objective of this study was to evaluate the effectiveness and safety of pseudomonas aeruginosa MSHA vaccine (PA-MSHA vaccine) used as an immune modulator. The study was carried out after the design of a non-blind, randomized controlled trial. The malignant lymphoma patients were divided into experiment group (45 cases) and control group (43 cases). Likewise were distributed the lung cancer patients: 44 cases in experiment group and 45 cases in control group. The two experiment groups received chemotherapy + PA-MSHA vaccine. The two control groups were given chemotherapy alone. The results showed that the clinical efficacy rate was 95.56% for the exp. group and 69.77% (P < 0.01) for the control group of the malignant lymphoma cases; the clinical efficacy rate was 59.09% for the exp. group and 42.22% (P < 0.01) for the control group of the lung cancer cases. The infection rate was 17.78% for the exp. group and 37.21% (P < 0.05) for the control group of the malignant lymphoma cases; the infection rate was 15.91% for the exp. group and 40.00% (P < 0.05) for the control group of the lung cancer cases. The comprehensive immune effectiveness rates for the exp. group and control group of the malignant lymphoma cases were 77.78% and 23.26% (P < 0.01) respectively. The comprehensive immune effectiveness rates for the exp. group and control group of the lung cancer cases were 84.09% and 35.56% (P < 0.01) respectively. Adverse reactions were found in 5 cases, of which, one was given medical care while the others automatically recovered. These results indicate that PA-MSHA vaccine, as a new kind of immune modulator, can improve the effectiveness of treatment for tumor with low adverse reaction rate. PMID- 12545826 TI - [Study on chemosensitivity assay in vitro in the peripheral blood lymphocyte and the tumor cells]. AB - In this study, the MTT method was used to test the sensitivity of the peripheral blood lymphocyte and the tumor cells of 35 patients with neuroglioma to 15 kinds of anti-cancer drugs. The results showed that the peripheral blood lymphocyte and the tumor cells were more sensitive to chemotherapeutic drugs Vm26 and TAX, and sensitive to DDP, Me-CCNU, EADM, ADM, MMC, HCPT, but not sensitive to MTX, ACR, VP-16, VCR and BLM. There were no statistical differences in the rate of sensitivity to the above-mentioned drugs between the peripheral blood lymphocyte and the tumor cells. These results prompt that the chemosensitivity test of the peripheral blood lymphocyte may take the place of the tumor cells for reference to choosing chemotherapeutic drugs in clinical practice. PMID- 12545827 TI - [Culture of endotheliocytes derived from guinea-pig common carotid artery and establishment of guinea-pig donor model]. AB - Since endotheliocyte strains of guinea-pig and inbred strains of guinea-pig are not easily obtained, it is impossible to ensure the consistence of antigenicity between endotheliocytes and donor organs when endotheliocytes are used as an inductor in the research of immunological tolerance. So it is necessary to establish the guinea-pig endotheliocyte-donor model. One side of common carotid arteries was removed and reversed. After collagenase digestion, culture of endotheliocytes was performed. And the survival time of guinea-pig was observed. Cultured cells were confirmed as endotheliocytes by the study of morphology, immunohistochemistry and electron microscopy. The two-month-survival of guinea pigs was 92% (103/112). The establishment of guinea-pig endotheliocyte-donor model can provide the foundation for the research of guinea-pig endotheliocyte and immunological tolerance. PMID- 12545828 TI - [Attenuation of hyperacute rejection from xenograft by reseeding of endothelial cell]. AB - The potential solution to palliate the critical shortage of suitable donor organs for transplantation may be xenogenic transplantation. However, hyperacute rejection (HAR) after xenotransplantation is the main problem in this process. Since the vascular endothelial cell of donor organ is the primary target cell in rejection, the replacement of endothelium in donor organs with endothelial cell from recipients themselves may be beneficial to the prevention of HAR. Discordant xenotransplantation model (guinea pig-to-rat) was adopted in this study. Firstly, endothelial cell from rat abdominal aorta was separated and cultured. Secondly, the guinea pig abdominal aorta of which the endothelium had been removed was cultured with the suspension containing rat endothelial cells (4 x 10(6)/ml). The viability of cultured vessel was assessed using light microscopy and transmission electron microscopy. The guinea pig vessel reseeded with rat endothelial cell was then examined by immuno-fluorescence staining assay to find out whether IgM and C3 in rat serum were bound to it after preincubation with rat serum. It was found that rat endothelial cells grew into the monolayer endothelium on the inner surface of guinea pig vessel with previous endothelial loss. IgM and C3 in rat serum did not deposit along the new endothelium of guinea pig vessel in the treated group as shown by immuno-fluorescence microscopy, whereas possive results were observed in the untreated normal guinea pig aorta. These findings indicate that donor vessel reseeded with endothelial cell from recipients undergoes less severe rejection and this technique may be very useful for the attenuation of HAR. PMID- 12545829 TI - [A study on the preservation of rat kidney with HX-III solution]. AB - This study was conducted to compare the effects of HX-III solution and UW solution on rat kidney transplantation. Inbred Sprague-Dawley rats weighing 200 280 g, were used in allotransplantation. The donors and recipients were of the same sex. The rats were randomly divided into control (UW solution) and experimental (HX-III solution) groups, and each group included 48- and 72-hour preservation subgroups. There were ten rats in each subgroup. Kidneys were flushed with UW solution or HX-III solution and stored at 4 degrees C for 48 or 72 hours. Then, kidney allotransplantations were performed. The results showed that the rates of survival were 100% and 90% in UW group for 48- and 72-hour cold storage, but those in HX-III were 100% and 60%, respectively. With 48-hour preservation, the maximum serum creatinine levels in UW group were similar to those in HX-III group (P > 0.05). With 72-hour preservation, the maximum serum creatinine levels in UW group were lower than those in HX-III group (P < 0.05). On the 14th day posttransplantation, the histological findings on Kidneys in the two groups for 48-hour cold storage were almost the same, but the grafted kidneys in HX-III group were damaged more seriously than those in UW group for 72 hour cold storage. On the 28th day posttransplantation, the histological findings on the grafted kidneys were basically normal in all groups. The authors concluded that HX-III solution could preserve rat kidney for 48 hours effectively. PMID- 12545830 TI - [Effects of sulpiride on morphine-induced reward and its relation to cyclic AMP levels in mice]. AB - This study examined the effects of sulpiride, a dopamine D2 receptor antagonist, on morphine rewarding properties and its relation to cyclic AMP levels in brain. The morphine-induced reward in mice was observed in a conditioned place preference test. Cyclic AMP was estimated by radioimmunoassay. The time of staying in morphine pairing compartment and the cAMP levels in brain of mice in the morphine group increased as compared with those in other groups (P < 0.05), but there was no significant difference in the time of staying in morphine pairing compartment and the cAMP levels in brain between the saline group and the sulpiride plus morphine groups (P > 0.05). These suggest that the inhibition of morphine-induced reward by sulpiride be due to the prevention of an increase in cyclic AMP levels in brain. PMID- 12545831 TI - [Effect of vasostomy on expression of Bcl-2 and Bax gene in rat spermatogenic cells]. AB - The objective of this study was to explore the change of Bcl-2 and Bax gene expression in rat spermatogneic cells after vasostomy. A rat model with vasoligation and vasostomy was set up and then used for detecting the protein expression of Bcl-2 and Bax gene at 4, 8, 12, 16 weeks after vasostomy by means of immunohistochemistry. Sixty rats were randomly distributed into three groups named vasoligation group (VG), vasostomy group(VOG) and sham-operation group(SOG) respectively. The results showed that the Bcl-2 protein expression in VOG group increased gradually and came to be significantly higher than that in VG group 8 weeks after vasostomy, and it's level was the same as that in SOG group at the 12th week. The bax protein expressions in VG and VOG groups were significantly higher than that in SOG group, but there was no difference between VOG and VG groups. In conclusion, the change of Bcl-2 protein expression observed in this study is indicative of the anti-apoptosis function of Bcl-2 gene in both the status of being vasoligated and vasostomized, whereas the consistent Bax protein expression before and often the repatency of the vas suggests further researches into its effect on apoptosis of spermatogenic cells. PMID- 12545832 TI - [The correlation of serum levels of insulin-like growth factors and intrauterine growth: report of a study in rats]. AB - This animal experiment was designed to investigate the relationship of the serum insulin-like growth factors (IGFs) with fetal intrauterine growth. The serum concentrations of IGF-I, -II in 21 fetal rats were measuerd and their relationships with fetal birth weight, length and tissue weights of brain, lung, liver were analysed respecively. The results showed that both of the serum concentrations of IGF-I, -II were positively correlated (P < 0.01) with fetal birth weight, length and tissue weights of brain, lung, liver respectively. These data suggest that both IGF-I and IGF-II may play an important role in fetal intrauterin growth and their decreased concentrations in serum may be factors contributing to fetal intrauterine growth retardation. PMID- 12545833 TI - [An experimental study on traumatic brain injury for inducing neuronal apoptosis in rats]. AB - This study was designed to explore the effect of traumatic brain injury and its mechanism for inducing neuronal apoptosis. The model of experimental traumatic brain injury was used. The rats were dispatched at different times(3 h, 12 h, 24 h, 48 h, 72 h) after traumatic brain injury, and the neuronal apoptosis was evaluated with microscope (HE staining), TUNEL method, flow cytometry, gel electrophoresis and immunohistochemistry assay. The results showed that the brain tissue neurons treated by traumatic brain injury underwent morphological changes of apoptosis, the DNA presented "ladder" break, the rate of neuronal apoptosis at different times ranged from 9.8% to 14.0%, but that of the control group was 1.7% and the expressions of related apoptosis genes(c-myc, fas and fasL) were increased. The findings of this study indicate that traumatic brain injury can induce neuronal apoptosis, and its molecular mechanism might be related to the expression of some apoptosis genes. PMID- 12545834 TI - [A comparative study on the capacities of different strains of Streptococcus sanguis for P-aminobenzoic acid production]. AB - This study was intended to compare the capacities of different strains of Strep, sanguis for P-Aminobenzoic acid (PABA) production. The synthesis of PABA during the growth of four strains of Strep. sanguis was measured by the reversed-phase high-performance liquid chromatographic method. The results showed that the concentrations of PABA synthesized by S. sanguis 10556, S. sanguis 10557, S. sanguis S34 and S. sanguis H7-4. Were 1.979 +/- 0.081 micrograms/ml, 1.383 +/- 0.193 micrograms/ml, 1.983 +/- 0.052 micrograms/ml and 1.032 +/- 0.229 micrograms/ml, respectively, and in term of PABA concentration, S. sanguis 10556 was significantly different from S. sanguis 10557 and S. sanguis H7-4; S. sanguis S34 was significantly different from S. sanguis 10557 and S. sanguis H7-4. No significant difference was found between S. sanguis 10556 and S. sanguis S34, nor between S. sanguis 10557 and S. sanguis H7-4, either. In conclusion, the method is simple, rapid and accurate. S. sanguis did synthesize PABA, and the difference in ability for PABA formation existed among the four strains of S. sanguis. This study is helpful to researches on the symbiosis between S. sanguis and S. muntans and to determination of their role in the microbial homeostasis of dental plaque. PMID- 12545835 TI - [Study on IGF-I regulation the proliferation of rat condylar chondrocytes in vitro]. AB - This study was aimed at the mechanism for the changes in the proliferation of cultured rat mandibular condylar chondrocytes. Flow cytometry was used to explore the changes of cellular DNA content and cell cycles of passaged condylar chondrocytes. The results showed that when the mandibular condylar chondrocytes were cultured with IGF-I for 24 hours and 96 hours, the cellular proliferation in the experimental group enhanced higher than that in the control group. It was implied that IGF-I may be a mitogen on rat mandibular condylar chondrocytes. The mitogenic effect of IGF-I had a lag period. PMID- 12545836 TI - [Study on biodegradation of phenols in river water]. AB - This study on the biodegradation of phenols (phenol, o-methylphenol, o chlorophenol and resorcinol) in river water was conducted by changing the main influential factors respectively. The results of experiment showed the phenols were greatly degradated in mild temperature, neutral pH and plenty of oxygen and the rank of the phenols biodegradation from great to small was resorcinol, phenol, o-methylphenol and o-chlorophenol. Presented in this paper is also a discussion on the biodegradation mechanism of phenols. PMID- 12545837 TI - [Effects of endothelin receptor antagonist on the hypoxic pulmonary hypertension]. AB - This investigation was made to elucidate the role of endothelin (ET) in hypoxic pulmonary hypertension and the preventing effects of BQ-123, an ETA receptor antagonist. Thirty male Wistar rats were divided into three groups and exposed to air, isobaric hypoxia or isobaric hypoxia plus BQ-123 for 3 weeks. The pulmonary artery pressure was measured by right cardiac catheterization. The plasma level of ET-1 was measured by RIA method. Histologic sections of the lungs were examined by a computerized image analyser. In hypoxic rats, the pulmonary artery pressure and the thickness of wall of arteriole were significantly increased, and right ventricular hypertrophy was developed. The plasma level of VEGF in rats treated with hypoxia (192.3 +/- 43.1 pg/ml) was significantly increased as compared with that of normal rats (128.2 +/- 28.1 pg/ml), P < 0.01. Chronic BQ 123 treatment prevented the developments of pulmonary hypertension, thickening of pulmonary arteriole and right ventricular hypertrophy induced by hypoxia. These result indicate that chronic hypoxia can result in hypoxic pulmonary hypertension and increased plasma level of ET-1, and the ETA receptor antagonist can prevent hypoxic pulmonary hypertension. PMID- 12545838 TI - [The changes of serum angiotensin converting enzyme activity in Type 2 diabetes and its vascular complications]. AB - This investigation was made with reference to the changes of serum angiotensin converting enzyme (SACE) activity in type 2 diabetes and its vascular complications. SACE activity was studied in 127 type 2 diabetic patients and 90 healthy persons by using a spectrophotometric assay. The results showed SACE activity was obviously higher in diabetic patients (459.51 +/- 175.85 U) than in healthy persons (321.14 +/- 121.27 U); SACE activity was significantly higher in type 2 diabetic patients with diabetic nephropathy (548.27 +/- 166.60 U) than in patients without diabetic nephropathy (383.2 +/- 139.00 U), but there was no difference between patients with microalbuminuria and macroalbuminuria; no statistical difference was detected in SACE activity between diabetic patients with diabetic retinopathy (465.64 +/- 178.93 U) and without retinopathy (449.07 +/- 170.04 U); SACE activity was not associated with the course of diabetes, blood pressure, blood lipid and blood glucose. These data suggest that raised SACE activity might only play a role in the initiation of type 2 diabetes and diabetic nephropathy, but not relate to the progress of diabetic nephropathy, the onset of diabetic retinopathy and hypertension. PMID- 12545839 TI - [Transthoracic three-dimensional echocardiography for determination of left ventricular mass in patients with dilated cardiomyopathy]. AB - This study was performed to compare left ventricular mass (LVM) measurement by one- (1D), two- (2D) and 3D echocardiography with magnetic resonance imaging (MRI) in patients with dilated cardiomyopathy (DCM). 36 patients with DCM underwent imaging by conventional 1D and 2D echocardiography as well as transthoracic 3D echocardiographic data acquisition. Also patients were imaged with cardiac MRI. LVM was determined. There was no significant correlation (r = 0.42, P > 0.05) for measuring LVM between 1D echocardiography and MRI in patients with DCM. A significant correlation was obtained between 2D echocardiography and MRI (r = 0.64, P < 0.01) and between 3D and MRI (r = 0.78, P < 0.01) in determination of LVM. Compared with 1D and 2D echocardiography, the 3D analysis achieved a significantly higher agreement with the MRI results (1D; 399.2 g, 2D: 285.9 g, 3D: 172.6 g versus MRI: 199.1 g). Interobserver variability was 5.1% for measuring LVM by 3D echocardiography. These results suggest that LVM of patients with DCM can be accurately determined by transthoracic 3D echocardiography and this method could be used in clinical practice. PMID- 12545840 TI - [A histopathologic and immunohistochemical study of 68 cases of hemangioblastoma]. AB - The aim of this study was to evaluate the histologic characteristics of hemangioblastoma (HB) and investigate the histogenesis of its stromal cells. Sixty-eight cases of hemangioblastoma were studied by light microscopy and thirty three cases were examined by immunohistochemical stainning. Forty-eight of the 68 patients were males and 20 were females. The average age was 37. Fifty-two cases (76.6%) had been present for two months to one year before the diagnosis was made. Sixty cases (88.2%) arose in the cerebellum. Fifty cases (73.5%) appeared as cystic nodules. Histologically, the 68 cases of HB were further categorized into three subtypes: typical (30 cases), cellular (21 cases) and reticular (17 cases). In 33 cases, the stromal cell were positive for NSE, but negative for F VIII, UEA-1, GFAP and EMA. In summary, there are three essential histologic categories of hemangioblastoma, and its stromal cells may well have the characters of neuroendocrine differentiation. PMID- 12545841 TI - [The relationship between arteriolar pathological changes and brain hemorrhage in primary hypertension cases]. AB - This study was intended to elucidate the relationship between arteriolar pathological changes and encephalorrhagia in the cases of primary hypertension. Gross anatomy and histology of brain were observed in autopsy of 45 cases of primary hypertension. Forty cases of arteriolar hyalinization including 26 cases of brain hemorrhage were found in those 45 cases of primary hypertension. The hyalinized arterioles were divided to two types. One is characterized by the narrow lumen of terminal arterioles, the other by the dilatant lumen of proximal arterioles. Hemorrhage always occurred in the latter type. Besides these changes, 3 cases of acute necrosis and 5 cases of atherosclerosis in arterioles were related to hemorrhage. These data suggest that brain hemorrhage of primary hypertension mainly occur in dilated and hyalinized arterioles, and some cases are probably related to acute necrosis or atheroscleromatous atrerioles. PMID- 12545842 TI - [Configurations of F-V curve and their transformation rule in COPD patients analysed with wave-speed theory]. AB - This study sought to analyse the mechanism on the formation of configurations of F-V curve and their transformation rule in COPD and cor pulmonale patients with wave-speed theory. The F-V curves were measured in 90 COPD patients (43 with chronic bronchitis, 47 with emphysema) and 31 complicated with cor pulmonale, all of them were in the ameliorated period. The indices selected were FVC, V75, V50, V25. Both the measured value and the measured value/predicted value (%) of all indices showed chronic bronchitis > emphysema > cor pulmonale. Never was there the concave type of normal humans in the F-V curves of COPD and cor pulmonale patients; however, the convex type was much increased in the curves and the hyperbolic type specific to COPD and cor pulmonale appeared. The mechanisms of formation for plateau, convex and hyperbolic types were analysed with the wave speed theory. In summary, the unifying rule for the transformation of F-V curve configuration from normal adolescents to old persons, smokers, COPD and cor pulmonale patients in various degrees can be outlined as, concave type normality convex type abnormality hyperbolic type. This implicates a unifying process of pulmonary functions from healthy state to weakness and from slight to serious abnormality. PMID- 12545843 TI - [Clinical study on interferon treatment of chronic idiopathic thrombocytopenic purpura]. AB - The efficacy and mechanism of interferon alpha-2a (IFN alpha-2a) were assessed in the treatment of chronic idiopathic thrombocytopenic purpura (cITP). 20 patients with cITP (treatment group) were treated with IFN alpha-2a 3MIU i.m. once a week for 8 weeks; 28 patients with cITP (control group) were treated with prednisone 1 mg/(kg.d) for 4 weeks. Blood platelet counts (BPC), megakaryocyte number, immunnological parameters, percent and absolute counts of reticulated platelets (RPs) and megakaryocyte colony formation units were observed before and after therapy. The results showed that the efficacy of IFN alpha-2a was better than that of corticosteroid, P < 0.05. The RPs% decreased and the platelet-producing megakaryocyte percentage increased from 11.43% to 33.19% significantly after IFN alpha-2a therapy, but there were no significant changes in immunnological parameters. These indicate that IFN alpha-2a is effective in treating cITP, the mechanism may be based on promoting the megakaryocyte development and activating the production of platelets. PMID- 12545844 TI - [Therapy for short stature in girls with Turner syndrome]. AB - This study was aimed to examine the growth-promoting effects of recombinant human growth hormone (r-hGH) and low dose of thyroid hormone on short stature in girls with Turner syndrome. 8 girls with Turner syndrome, at the chronological age of 8.5-14.5 years (12.3 +/- 2.1 years) and bone age of 8.3-12.7 years (9.8 +/- 1.7 years) were observed. The maximum serum GH levels in 3 patients out of 5 with provocative testing available were > or = 10 ng/ml (10.0, 10.0 and 15.52 ng/ml, respectively), and in the other 2 of the 5 patients were < 10 ng/ml (9.3 and 9.5 ng/ml). Thyroid function including T3, T4 and TSH were measured in 5 patients. The TSH in one patient was slightly higher; the T3, T4 and TSH in other patients were normal. All patients were treated with r-hGH 1.0 IU/(kg.wk) and thyroideum 15 mg/d for 6-36 months. The results showed that the growth rate after treatment was remarkably higher than that before treatment (2.2 +/- 1.1 cm/yr). The growth rates in the first 6 month, the first year, the second year and the third year were 7.9 +/- 1.6 cm/yr, 7.5 +/- 1.2 cm/yr, 6.9 +/- 0.2 cm/yr and 5.0 +/- 0.2 cm/yr, respectively. Height standard deviation score (HSDS) increased 1.3 +/- 1.1 SD, compared with that pretreatment. The annual bone age growth rate after treatment was 0.6 +/- 0.2 yr/yr (0.43-0.8 yr/yr). These data indicated that r-hGH and low dose of thyroid hormone had remarkable effects of growth-promoting in the girls with Turner syndrome. PMID- 12545845 TI - [Effectiveness of recombinant human growth hormone treatment for severe burn injury]. AB - In order to investigate the effectiveness and safety of recombinant human growth hormone(rhGH) treatment for severe burn injury, we designed a randomized prospective study. The patients in rhGH group were given rhGH in a dose of 0.5 IU/(kg.d) subcutaneously between days 3-17 post-burn. Indices about protein metabolism, wound healing, hepatic and renal function and blood were measured regularly. Patients were monitored until their wound healed sufficiently so that they could be discharged for rehabilitation. The results showed: all patients survived; the rhGH group had less weight loss, increased level of serum albumin and shorter healing time of deep partial-thickness burn and donor site. The hospital stay time decreased from the control value of 45 +/- 17 days to 37 +/- 12 days for rhGH treated patients, i.e. a significant improvement. Resistant hyperglycemia was present in 2 patients in the rhGH group and disappeared after transient insulin administration. This study demonstrates that rhGH treatment for severe burn injury has obvious beneficial effects with slight side-effects. PMID- 12545846 TI - [Value of intestinal decontamination by traditional Chinese medicine-X in the prevention of bacterial translocation complicated by severe acute pancreatitis in rats]. AB - This study was designed to evaluate the efficacy of traditional Chinese Medicine X in preventing the necrotic infection of the pancreas in severe acute pancreatitis (SAP). Sixty rats were randomly divided into five groups with 12 rats in each one: (1) normal control, (2) SAP + 0.9% normal saline (1 ml x 100 g 1 x 24 h-1), (3) SAP + gentamycin (2000 u x 100 g-1 x 24 h-1), (4) SAP + TCM-X (1.0 g x 100 g-1 x 24 h-1), and (5) SAP + gentamycin (2000 u x 100 g-1 x 24 h-1) + TCM-X (1.0 g x 100 g-1 x 24 h-1). The medicines were given by way of gastrotube, once every 24 hours, twice in all. Pancreatitis was induced by a single intraperitoneal injection of 500 mg.100 g-1 of L-arginine. Serum endotoxin were observed and the clone forming units from mesenteric lymphnode and pancreas were obtained after 48 hours treatment. 96 hours after the experiment, the bacteria found in the mesenteric lymphnodes and pancreas in groups three, four and five were reduced as compared to that in group two; the levels of serum endotoxin were reduced, too. These data indicate that TCM-X and gentamycin in decontamination by way of gastrotube are effective in preventing bacterial translocation complicated by SAP, and the effect of TCM-X is stronger than that of gentamycin. PMID- 12545847 TI - [Electron spin resonance studies on the oxidative denaturation of hemoglobin of blood preserved at 20 degrees C]. AB - In order to further understand the injurious change of erythrocytes during blood preservation at room temperature, we studied the denaturation of oxidative hemoglobin of blood preserved at 20 degrees C using electron spin resonance (ESR) technique. The results showed: when blood was preserved at 20 degrees C, there was ESR absorption of low spin hemichrome (g = 2) during the 204-hour preservation period; when the blood was preserved at 4 degrees C after it had been preserved at 20 degrees C for 24 hours, there were ESR absorption of the high spin methemoglobin (g = 6) and ESR absorption of low spin hemichrome (g = 2) during the 40-day preservation period. This study demonstrates that ESR absorption will increase when preservation time increases. When blood is preserved at 4 degrees C, no ESR absorption will emerge during 40 days' preservation. PMID- 12545848 TI - [Nonsyndromic cleft lip with or without cleft palate in Chinese population: analysis of 3766 cases]. AB - A retrospective study was conducted to evaluate the variation trend in the incidence and epidemiological characteristics of nonsyndromic cleft lip with or without cleft palate in Chinese population. From 1988 to 1992, data were collected from more than 500 hospital-based units involved in a monitoring program and were analyzed by the National Center for Birth Defects Monitoring. The monitored subjects were the hospitalized births from 28 weeks gestation to a period within 7 days after delivery, which included 3246 408 perinatals and 3766 cases. The total incidence of nonsyndromic cleft lip with or without cleft palate was 11.6/10,000; the total incidence of celft lip alone was 3.8/10,000 and that of cleft lip with cleft palate was 7.8/10,000; no significant variation was noted in the annual incidence from 1988 to 1992. The incidence in males was 13.3/10,000, and 9.5/10,000 in females; there was a significant difference (P < 0.01). Of all the 3766 cases, cleft lip with cleft palate accounted for 67%, unilateral cleft lip 28% and unilateral cleft lip with cleft palate 43%. Left cleft lip and left cleft lip with cleft palate constituted 21% and 29% respectively. The sex-ratio was 1.6:1 for nonsyndromic cleft lip with or without cleft palate. It could be concluded that the incidence of nonsyndromic cleft lip with or without cleft palate showed no variation trend in China. Although no difference was found between urban and rural areas, the susceptibility to the defect was higher in males than in females. Cleft lip with cleft palate was more common than other types. There was a predominance of unilateral defects, of which most were on the left side. PMID- 12545849 TI - [Immobilization osteoporosis and fracture in children]. AB - This study was intended to enhance the diagnostic vigilance of immobilization osteoporosis with fracture and explore its prevention and treatment. Thirteen children with the hip disease underwent one to nine months of plaster cast after operation. A retrospective analysis was made on these 13 patients with fracture and immobilization osteoporosis. The result showed that disuse fracture occurred in fifteen places of their femora. All of these cases were cured by non-operative and drug treatment. These data suggest that for the cases of immobilization osteoporosis it is important to prevent fracture by means of early diagnosis, proper drugs, early function exercise, appropriately shorter period of fixation and the avoidance of trauma. PMID- 12545850 TI - [Study on fluorimetry of aluminum quaternary complex and analytical application]. AB - This study was conducted to seek an easy-to-do method for determination of aluminum in biological and food samples. The fluorescent reaction of aluminum forming quaternary complex with fluoride, Ferron and CTMAB in the buffer solution of ammonium acetate (at pH 6.0) was investigated. The conditions of the reaction and the pretreatment of samples were optimized. In addition, the aluminum contents of hair and food samples was detected. A good linear relationship between fluorescence intensity and aluminum concentrations from 0.002 microgram/ml to 0.20 microgram/ml was observed. The detection limit was 1.0 x 10( 3) micrograms/ml. The spiked recoveries were in the range of 88.66%-112.61% with relative standard deviation varying from 1.87% to 2.56%. This method has characters of good sensitivity, selectivity and the ability against interference, it can be of wide application. PMID- 12545851 TI - [The establishment of a computerized growth prediction system]. AB - Natural craniofacial growth plays an important role in the etiology of malocclusion. So, craniofacial growth prediction is very necessary in orthodontics. The present author have developed a new-typed computerized craniofacial growth predition system, which was designed in the general sense of craniofacial growth. According to a unique method, the system could automatically predict of craniofacial growth to different craniofacial growth pattern. It can be used as an auxilliary tool in clinical, teaching and researches in orthodontics. PMID- 12545852 TI - [Determination of blood glucose by chemiluminescent flow-injection analysis with immobilized glucoxidase column]. AB - Luminol-hemin system was used for chemiluminescent flow-injection analysis with immobilized glucoxidase(GOD) column to determine blood glucose. The method is fast, simple and sensitive. Blood samples were diluted to 1:1000 and pumped into the flow-injection analyzer for direct detection. The sampling frequency was 30 samples/h, with a detection limit of 7.2 x 10(-9) mol/L(1.30 micrograms/L). A column packed with 36 milligrams of CPG could be used nearly 200 times. The determination results of glucose in serum by this method were in good agreement with those obtained by the routine GOD spectrophotometric method (correlation coefficient r = 0.90, P > 0.05). Recoveries of this method were 94.0%-107.2%. PMID- 12545853 TI - [Research progress in physiological functions of selenoenzyme and other selenocompounds]. AB - Selenium (Se) is an essential trace element for animals. Selenocysteine (Sec), the 21st aminoacid, is a component of selenoproteins and has been founded in the active center of selenoenzymes. The functions of Se within the body have been primarily shown in the forms of selenoproteins, especially selenoenzymes. Incorporation of selenocysteine occurs on the basis of genetic expression and Se is the only trace element under direct genetic control. Recently, findings have shown that Se and selenocompounds conducted many other potential functions such as protection against inflammatory factors, inhibition of protein kinase C (PKC), stimulation of MAP kinase (mitogen activated protein kinase/myelin basic protein kinase) and S6 kinase (ribosomal S6 protein kinase), regulation of the immune system and interaction with other elements and vitamins etc, suggesting that the roles of Se in human health may be more diverse than previously suspected. PMID- 12545854 TI - [Advancement in mechanisms of long-term potentiation]. AB - Synaptic plasticity in mammalian brain is one of the most widely studied topics in neuroscience over the last decade. Long-term potentiation (LTP), mainly involving post-synaptic mechanisms, is a reflection of neural plasticity. Study on the mechanisms of LTP was formerly focused on the properties of NMDA receptor and the intracellular cascade of reaction after activation of the receptor. However, the definition of "silent synapses" with only NMDA receptors without AMPA receptors was frequently referred in LTP study, showing the importance of AMPA receptor in post-synaptic mechanism of LTP. PMID- 12545855 TI - [Molecular candidates for capacitative calcium entry channel]. AB - The emptying of intracellular calcium store generates a putative signal that induces the opening of the store-operated calcium channel (SOC) at the level of the cell membrane, also known as the capacitative calcium entry (CCE), or the calcium release-activated calcium channel (CRAC), which allows entry of calcium into the cells from the extracellular space. Drosophila transient receptor potential (trp) and trp-like(trpl) genes have been identified and reported to code cation channels(TRP and TRPL). The mechanism of coupling between store depletion and channel opening is unknown. Three fundamental mechanisms have been proposed for transmitting the signal from intracellular stores to the plasma membrane: (1) a diffusible message; (2) protein-protein interactions; and (3) vesicle secretion. This review focuses primarily on the recent studies on the TRP homologues, the possible mechanisms and electrophysiological phenotypes of capacitative calcium entry. PMID- 12545856 TI - [Alzheimer's disease related-genes and apoptosis]. AB - Alzheimer's disease (AD) is the most common cause of dementia in the elderly population. The occurrence of dementia correlates best with neuronal apoptosis. All the proteins (APP, PS1, PS2) encoded by AD-related genes are involved in regulating neuronal apoptosis. On the other hand numerous apoptosis-regulating factors have been identified to participate in AD neurodegeneration. Detailed studies on this field are of important significance in the understanding the pathogenesis and the therapeutic measures for AD. This review will discuss the interrelation and interaction between AD-related genes and apoptosis. PMID- 12545857 TI - [Gene therapy of hypertension]. AB - Hypertension is a disease caused by multiple genes. The traditional treatments have many defects. A gene therapy has been proposed where antisense oligonucleotide (AS ODN) is developed to specifically block the expression of hypertension candidate genes. It is of long-term effect, high efficiency and non toxicity. Preliminary results are encouraging, but much work needs to be done before gene therapy could be applied to humans. PMID- 12545858 TI - [Progress in the studies of CD34-hematopoietic stem cell]. PMID- 12545859 TI - [Hematopoiesis and immunity regulation mediated by histamine receptors]. PMID- 12545860 TI - [Novel analgesics: neuronal nicotinic acetylcholine receptor agonists]. PMID- 12545861 TI - [The biology and related disease of voltage-gated chloride channel (CIC chloride channel)]. PMID- 12545862 TI - [The composition and biological role of ubiquitin-proteasome pathway]. PMID- 12545863 TI - [Dimerization of G-protein coupled receptor and its influence on receptor mediated signal transduction]. PMID- 12545864 TI - [Research progress in the cardiovascular action of estrogens]. PMID- 12545865 TI - [Modulation of the autonomic nervous system cardiac ion channels]. PMID- 12545866 TI - [PSD-95 integrates the signal transduction mediated by NMDA receptors]. PMID- 12545867 TI - [Endoplasmic reticulum and Alzheimer's disease]. PMID- 12545868 TI - [Progress of the all models of Alzheimer's disease and their applications in pharmacological study]. PMID- 12545869 TI - [Advances in the study of glial fibrillary acidic protein]. PMID- 12545870 TI - [Molecular origin of basal metabolic rate in mammals]. PMID- 12545872 TI - [The relation of histone acetylation/deacetylation and DNA methylation]. PMID- 12545871 TI - [A study of controlling mechanism on mitochondrial respiratory chain function]. PMID- 12545874 TI - [Event-related functional magnetic resonance imaging]. PMID- 12545873 TI - [The application and prospect of SELEX technology]. PMID- 12545875 TI - [Gap junctions and cardiovascular diseases]. PMID- 12545876 TI - [Transplantation of peripheral nerves and their tissue constituents to repair axotomized retinal ganglion cells in adult mammals]. AB - We focus on how peripheral nerves or their tissue constituents including Schwann cells, fibroblasts and neurotrophic factors are used to overcome the unfavorable extrinsic CNS environment and upregulate the intrinsic growth potential of injured neurons for the enhancement of neuronal survival and axonal regeneration of axotomized retinal ganglion cells in adult mammals. PMID- 12545878 TI - [Sympathetic-sensory coupling in neuropathic pains]. AB - Peripheral tissue and nerve injury lead to chronic pain symptoms, including spontaneous pain, allodynia and hyperalgesia. Sympathetic nervous system involves in pathological changes above, either through development of abnormal sympathetic function or through its effects on abnormally functioning afferent nerves, thereby to cause many neuropathic pains. This review focuses on the progress at present of sympathetic-sensory coupling and its receptors, intracellular and neural mechanisms. PMID- 12545877 TI - [A review of brain aromatase]. AB - Aromatase catalyzes the conversion of androgen to estrogen. This enzyme has been primarily localized in the neurons of specific areas of limbic system and hypothalamus of the brain. Astrocytes may also express this enzyme. Studies show that the gene expression of aromatase is driven by multiple tissue-specific exons, and the effective concentration of brain estrogen depends on the expression of brain aromatase. The locally produced estrogen plays an important role in the regulation of synaptogenesis, density of dendritic spine and the expression of neurotrophic factors and/or their receptors. Estrogen can also protect brain cells from the damages of neurotoxins and can greatly improve the deficits of learning and memory resulted from neurodegenerative disorders such as Alzheimer's diseases. PMID- 12545879 TI - [The hypothalamic suprachiasmatic nucleus and pineal gland in the circadian rhythmic organization of mammals]. AB - The hypothalamic suprachiasmatic nucleus (SCN) is a most important circadian pacemaker, which controls physiological and behaviour rhythm in mammals. SCN owns a intrinsic rhythm itself, and is entrained by photoperiodic signal and some endogenous chemical substances. Melatonin (MEL) is secreted by pineal gland, which is regulated by SCN. MEL triggers the second and third message systems, and regulates SCN circadian activity through high affinity MEL receptor within SCN. This regulation is time-sensitive. PMID- 12545880 TI - [Role of phospholipase D in phagocytosis and exocytosis]. AB - Mammalian phospholipase D(PLD) is a very important enzyme in the cellular signalling pathways. More and more lines of evidences suggest that PLD may be pivotal on multiple specialized steps in receptor-mediated phagocytosis and exocytosis. In this review, we will explore the recent advances on the role and the mechanism of PLD in phagocytosis and exocytosis. PMID- 12545881 TI - [Effect of BSAP/Pax-5 on the development, proliferation and differentiation of B lymphocyte cell]. AB - BSAP, a B cell lineage-specific activator protein, is a nucleus transcription factor and is encoded by the Pax-5 gene. It is a critical modulator of B cell development, proliferation and differentiation. BSAP also influences B cell immunoglobulin secretion at later stages of B cell differentiation. PMID- 12545882 TI - [Advances in the research of penile erection and erection dysfunction]. AB - Recent advances in the research on erectile dysfunction (ED) has more clearly explained the mechanism of penile erection, which is a hemodynamic process involving relaxation of corpus cavernosum smooth muscle and related arterials, and NO-cGMP signaling pathway proven to play an important role on modulating the relaxation of corpus cavernosum smooth muscle. Studies on NOS and PDEs not only provide strong evidence for the clinical treatment of ED, but also provide the chance to develop Sildenafil. Although the gene therapy for ED is still in the laboratory stage by now, it may be one of the effective clinical therapies for ED in future. PMID- 12545883 TI - [Peroxynitrite-mediated pulmonary vascular injury induced by endotoxin and the protective role of cholecystokinin]. AB - This study was to investigate the mediation of peroxynitrite(ONOO-) in pulmonary vascular injury induced by lipopolysaccharide(LPS), a major component of bacterial endotoxin, and the protective effect of cholecystokinin octapeptide(CCK) and its underlying mechanism. The generation of ONOO- in rat lungs was induced by administration of LPS (5 mg/kg i.v.). Exogenous ONOO- led to increase in microvascular permeability and severe pathologic changes of rat lungs. The exposure of isolated pulmonary artery segment to ONOO- resulted in abnormal reactivities similar to those induced by LPS. ONOO- caused weak relaxation which was negatively regulated by endothelial cells. The significantly increased production of endogenous ONOO- in cultured BPAEC elicited by LPS might mediate cytotoxic effects of LPS. The protection of CCK against the effects of LPS in isolated pulmonary artery or BPAEC was mediated by CCK receptors and related to the inhibition of ONOO- generation. These data indicated that scavenging ONOO- or decreased in ONOO- generation provided a novel therapeutic strategy for alleviation of LPS-induced pathologic process such as acute lung injury, and that CCK may be an endogenous cytoprotective factor for promising practice. PMID- 12545884 TI - [Advance in stem cell research and clinical application]. PMID- 12545885 TI - [Stem cells in the adult mammalian central nervous stem]. PMID- 12545886 TI - [The expression and biological effects of stem cell factor and its receptor in nervous system]. PMID- 12545887 TI - [Calcium signaling modulates gene expression]. PMID- 12545888 TI - [Nicotinic acetylcholine receptors in central nervous system: too many properties and too few functions]. PMID- 12545889 TI - [Myeloid defensins: an update review]. PMID- 12545890 TI - [The recent advances in the study of a novel cell membrane skeleton protein- adducin]. PMID- 12545891 TI - [Research progress in the oncostatic mechanism of melatonin]. PMID- 12545892 TI - [The new progress in the study of calcitonin gene-related peptide]. PMID- 12545893 TI - [Physiological function of kainate receptors]. PMID- 12545894 TI - [G-protein-coupled inwardly rectifying potassium channel in central nervous system]. PMID- 12545895 TI - [The actions of glucocorticoid receptor and 11 beta-hydroxysteroid dehydrogenases in the brain]. PMID- 12545896 TI - [The related genes involvement in long-term potentiation]. PMID- 12545897 TI - [Progress in the study of orphan nuclear receptor]. PMID- 12545898 TI - [Strategies and methods of cloning disease genes]. PMID- 12545900 TI - HCA Richmond Hospitals' new marketing strategy a winning plan. AB - HCA Richmond Hospitals, a five-hospital system in Richmond, Va., is positioning itself as a winner in a highly competitive, healthcare-saturated market since overhauling is marketing strategy a little over a year ago. The marketing strategy enables individual hospital to target their own unique constituencies. "Understanding the intricate marketing dynamics of hospital systems is today of critical importance and equal complexity," said Tony Bejamin, principal of Oxygen Advertising Inc., New York, the agency that remodeled HCA Richmond Hospitals' marketing strategy. PMID- 12545901 TI - Spartanburg attracts moms-to-be with a variety of media. Campaign pieces have won numerous awards. AB - Spartanburg Regional Healthcare System, based in Spartanburg, S.C., uses a variety of media to attract moms-to-be and other women to its many services. The award-winning campaign was cited by The National Federation of Press Women with nine awards and a First Place Sweepstakes Award. PMID- 12545902 TI - St. John Health integrating new corporate identity, brand. Brand/logo to be rolled out over two-year period. AB - St. John Health, Warren, Mich., is integrating a new corporate identity and brand strategy for its network of nine wholly-owned and two affiliated hospital, along with more than 100 physician offices and specialty centers in southeast Michigan. "A new identity is a great rallying cry. It automatically says. 'We have a new mission. We have a new system. We are reaching new people,'" said Eunice O'Loughlan, VP, corporate communications for St. John Health. PMID- 12545903 TI - Georgia Hospital Association leads long-term staff recruiting effort. Reaches wide target audience through innovative media use. AB - The Georgia Hospital Association (GHA), faced with a critical healthcare workforce shortage, created a long-term response to the problem by creating a Workforce Shortage Media Campaign. Its single purpose was to encourage Georgians to pursue careers in healthcare. The four-week campaign took six months to develop and included print, radio and television. PMID- 12545904 TI - Beebe isn't 'boo boo' any more. In a 'readers' market, magazine helps change public perception. AB - Beebe Medical Center, Lewes, Del., was referred to by many of its neighbors as "Booboo Hospital," but all that changed when Sharon Perry, director of public relations, created the Beacon, a magazine ideally suited to her market that is largely retirees and people ages 55 and older. PMID- 12545905 TI - The Methodist Hospital Web site comprehensive, user friendly. AB - The Methodist Hospital, Houston, has an award winning Web site that truly deserves its wide recognition. This comprehensive Web site, www.methodisthealth.com, is taking on a new look with the beginning of the new year. It's still loaded with information that's easily accessed in both English and Spanish. PMID- 12545906 TI - A question of integrity. Federal prosecutors question Tenet's compliance with integrity agreement in a $323 million False Claims Act lawsuit. AB - Tenet Healthcare Corp.'s carefully groomed reputation as a kinder and cleaner for profit hospital chain alternative took another hit last week when the U.S. Justice Department filed a $323 million lawsuit against the company. The False Claims Act suit alleges Tenet illegally submitted thousands of false claims to maximize revenue. Lawyer Michael Ruggio says Tenet's troubles are far from over. PMID- 12545907 TI - It's good to be king again. PMID- 12545908 TI - As Congress' priorities shift, hopes for healthcare dim. AHA, AMA sponsor 'fly ins' to plead their cases for new round of hospital, physician funding. PMID- 12545909 TI - Hospital care fuels spending. PMID- 12545910 TI - Bled dry. Fiscal woes threaten supply data from National Blood Data Resource Center. PMID- 12545911 TI - IOM report sets priorities. PMID- 12545912 TI - Blues caught in bind. Proposed Pa. tax on insurers' reserves clashes with association's policy. PMID- 12545913 TI - Fund raising hits a privacy barrier. HIPAA rule will force healthcare organizations to rethink development outreach. PMID- 12545914 TI - Web survey. December results: 'looking forward to 2003'. PMID- 12545915 TI - A community solution. Local health centers are credited with keeping the safety net strong. President Bush wants them to play an even bigger role. PMID- 12545916 TI - Out for blood. Although automation and 'bloodless' surgical techniques are helping to conserve the nation's blood supply, the best way to stay well-stocked remains a steady stream of donors. PMID- 12545918 TI - 2004 decision process model approved for development. PMID- 12545917 TI - Accreditation timeline charts transition from old process to new model. PMID- 12545919 TI - Heikkinen-Komonen architects create a sense of community for scientists at the Max Planck Institute in Dresden. PMID- 12545920 TI - The best-laid incentive plans. AB - Hiram Phillips couldn't have been in better spirits. The CFO and chief administrative officer of Rainbarrel Products, a diversified consumer-durables manufacturer, Phillips felt he'd single-handedly turned the company's performance around. He'd only been at Rainbarrel a year, but the company's numbers had, according to his measures, already improved by leaps and bounds. Now the day had come for Hiram to share the positive results of his new performance management system with his colleagues. The corporate executive council was meeting, and even CEO Keith Randall was applauding the CFO's work: "Hiram's going to give us some very good news about cost reductions and operating efficiencies, all due to the changes he's designed and implemented this year." Everything looked positively rosy--until some questionable information began to trickle in from other meeting participants. It came to light, for instance, that R&D had developed a breakthrough product that was not being brought to market as quickly as it should have been--thanks to Hiram's inflexible budgeting process. Then, too, an employee survey showed that workers were demoralized. And customers were complaining about Rainbarrel's service. The general message? The new performance metrics and incentives had indeed been affecting overall performance--but not for the better. Should Rainbarrel revisit its approach to performance management? Commentators Stephen Kaufman, a senior lecturer at Harvard Business School; compensation consultant Steven Gross; retired U.S. Navy vice admiral and management consultant Diego Hernandez; and Barry Leskin, a consultant and former chief learning officer for Chevron Texaco, offer their advice in this fictional case study. PMID- 12545921 TI - Moving mountains. AB - What could be more fundamental to management, or more difficult, than motivating people? After all, a manager, by definition, is someone who gets work done through others. But how? A typical recipe for motivation calls for a mixture of persuasion, encouragement, and compulsion. Yet the best leaders, we suspect, need no recipe: They get people to produce great results by appealing to their deepest drives, needs, and desires. And so we discovered when we asked a dozen of the world's top leaders to describe how they each met a daunting challenge in motivating an individual, a team, or an organization. Their answers are as varied as human nature. Some of the leaders appeal to people's need for the rational and the orderly: Mattel's Robert Eckert emphasizes the reassuring power of delivering a consistent message, and HP's Carly Fiorina focuses on facing hard truths on setting step-by-step goals. Some, like celebrated oceanographer Robert Ballard, Pfizer CEO Hank McKinnell, and BP America president Ross Pillari, see the powerful motivating effects of asking people to rise to difficult challenges. Others focus more on the human spirit, appealing to the desire to do something, as BMW's Chris Bangle puts it, "rare, marvelous, and lasting." And quite a few inspire through example, as Dial chairman Herb Baum did when he donated $1,000 from his bonus to each of the company's 155 lowest-paid people. "If you draw the line on your own greed, and your employees see it," he says, "they will be incredibly loyal and perform much better for you." And he has the numbers to prove it. "Right now," he adds, "we're experiencing our lowest level of attrition in 11 years, and we're tracking toward another banner year because people are happy." PMID- 12545922 TI - Beyond empowerment: building a company of citizens. AB - We live in a knowledge economy. The core assets of the modern business enterprise aren't its buildings, machinery, and real estate, but the intelligence, understanding, skills, and experience of its employees. Harnessing the capabilities and commitment of knowledge workers is arguably the central managerial challenge of our time. Unfortunately, it is a challenge that has not yet been met. Corporate ownership structures, governance systems, and incentive programs--despite the enlightened rhetoric of business leaders--remain firmly planted in the industrial age. In this article, the authors draw on history to lay out a model for a democratic business organization suited to the knowledge economy. Some 2,500 years ago, the city-state of ancient Athens rose to unprecedented political and economic power by giving its citizens a direct voice and an active role in civic governance. The city's uniquely participative system of democracy helped unleash the creativity of the Athenian people and channel it to produce the greatest good for society. The system succeeded in bringing individual initiative and common cause into harmony. And that is precisely the synthesis today's companies need to achieve if they're to realize the full power of their people and thrive in the knowledge economy. The Athenian model of organizational democracy is just that--a model. It does not provide a simple set of prescriptions for modern managers. What it offers is a window into how sizable groups of people can, in an atmosphere of dignity and trust, successfully govern themselves without resorting to a stifling bureaucracy. PMID- 12545923 TI - How to motivate your problem people. AB - Managers who motivate with incentives and the power of their vision and passion succeed only in energizing employees who want to be motivated. So how do you motivate intractable employees--the ones who never do what you want and also take up all your time? According to Nigel Nicholson, you can't: Individuals must motivated themselves. Nicholson advocates a method that turns conventional approaches to motivation upside down. Instead of pushing solutions on problem employees, the manager should pull solutions out of them by creating circumstances in which the employees can channel their motivation toward achievable goals. That means addressing any obstacles-possibly even the manager's own demotivating style--that might be hindering the employees. The author's method demands that a manager take charge of a difficult situation and resolve it. An investment of time is required, but it will bring the manager to a resolution sooner than other means would. Using detailed examples, Nicholson walks the reader through his method, pointing out potential pitfalls along the way. First, the manager creates a rich picture of the problem person. Second, the manager exercises flexibility and reframes goals so that the employee can meet them. Third, in a carefully staged, face-to-face conversation, the manager meets with the problem employee on neutral ground. Whether a problem is solved or simply resolved, the payoffs from using this method extend beyond the specific employees who have been difficult to motivate. Besides increasing a manager's chances of motivation problem people, the method can inspire an entire team by signaling that the organization deals with difficult people rather than discarding them. PMID- 12545924 TI - What's wrong with executive compensation? A roundtable moderated by Charles Elson. AB - The value that many superpaid CEO superstars supposedly created has largely disappeared, and the likelihood that it will be recovered anytime soon seems remote. On top of that, a good number of top executives treated their companies like ATMs, awarding themselves millions of dollars in corporate perks. It's hard to dispute the idea that executives were corrupted by the sums of money dangled in front of them. What's wrong with executive compensation, and what can we do about it? HBR and the University of Delaware's Center for Corporate Governance convened a round-table of compensation experts last October on the university's campus in Newark, Delaware. The 12 panelists, from CEOs to investors, from the professionals who advise them to a chief justice who rules on their disputes, provided an extraordinary diversity of viewpoints. The panelist began by debating ways to align the interests of the senior executives with the long-term interests of the company-weighing the relative benefits of stock options versus stock grants, for instance. But the discussion expanded to cover broader questions of corporate governance and company values. "The main reason compensation increases every year is that most boards want their CEO to be in the top half of the CEO peer group," said Ed Woolard,Jr., a former CEO of DuPont. And compensation lawyer Joe Bachelder pointed out the danger of structuring pay in such a way that it dampens risk taking among executives. It was a lively and wide-ranging discussion of one business's most pressing issues. PMID- 12545925 TI - One more time: how do you motivate employees? 1968. PMID- 12545926 TI - Pygmalion in management. 1969. PMID- 12545927 TI - Management by whose objectives? 1970. PMID- 12545928 TI - Power is the great motivator. 1976. PMID- 12545930 TI - A better way to eat. PMID- 12545931 TI - Rx: two martinis a day. PMID- 12545929 TI - Stacking up the perfect diet. PMID- 12545932 TI - All about vitamin pills. PMID- 12545934 TI - Dissecting the diets. PMID- 12545933 TI - Feeding the aging brain. PMID- 12545935 TI - New ideas about halting diabetes. Are dramatic changes in lifestyle an effective way to deal with the disease? PMID- 12545936 TI - Get up and get moving. PMID- 12545937 TI - Concentrating on the body's 'core'. PMID- 12545938 TI - A little more willpower can change your life. PMID- 12545939 TI - Grown-up and uninsured. PMID- 12545940 TI - Modern concepts of functional electrical stimulation. PMID- 12545941 TI - Functional electrical stimulation for a dropped foot. AB - The purpose of this collective review is to study the techniques, usage, methods, and clinical results of functional electrical stimulation applied to the peroneal nerve in the treatment of dropped foot. When stimulation is applied through surface electrodes, clinical reports have documented the therapeutic and orthotic benefits of functional electrical stimulation. Clinical trials are now being undertaken in which implantable electrodes are being used to stimulate the peroneal nerve in patients with dropped foot. PMID- 12545942 TI - Functional electrical stimulation cycle ergometer exercise for spinal cord injured patients. AB - The purpose of this collective review on functional electrical stimulation (FES) cycle ergometer training is to describe the pathologic effects of spinal cord injury (SCI) and the structure and function of the FES cycle ergometers, which reverse the devastating systemic and life-threatening effects of SCI. The pathophysiologic consequences of SCI include diminished cardiopulmonary and circulatory function as well as lower extremity muscle atrophy and bone mass reduction. Clinical studies have demonstrated that the two FES cycle ergometers offer promise in reversing these devastating consequences of SCI, which can shorten patients' lives. On the basis of this collective review, it is recommended that all patients with SCI have the benefits of this potentially life sustaining clinical modality. PMID- 12545943 TI - Functional electrical stimulation in tetraplegic patients to restore hand function. AB - The purpose of this collective review is to describe a new form of functional electrical stimulation called neuroprosthesis. This unique technology has been devised to produce lateral pinch and palmar grasp in persons with C5 and C6 motor level spinal cord injuries. This neuroprosthesis includes external as well as implanted components. First, a receiver is surgically implanted into the patient's chest above a pectoralis major muscle. The receiver stimulator is then connected to 8 surgically implanted epimysial or intramuscular electrodes. Restoration of upper extremity function can greatly improve the lives of people affected with tetraplegia. When contralateral shoulder movements trigger an external transmitting coil, it sends a radio wave impulse to the stimulator inducing contraction of the muscles. Many tetraplegics are regaining hand function using implanted functional electrical stimulation. One major limitation is that the key muscles to be stimulated may have lower motor neuron damage, but this obstacle has been successfully overcome using surgical modifications of the biomechanics of the hand. PMID- 12545944 TI - Functional electrical stimulation of bladder and bowel in spinal cord injury. AB - The purpose of this collective review is to examine the use of functional electrical stimulation for incontinence. The Finetech-Brindley bladder system enhances voiding through stimulation via electrodes implanted around the ventral sacral roots. Detrusor hyperreflexia is eliminated through complete dorsal rhizotomy, which results in loss of reflex defecation and reflex erection/reflex lubrication. Consequently, a new system is being devised in which functional electrical stimulation for incontinence in spinal cord injury can be achieved without dorsal rhizotomy. PMID- 12545945 TI - Functional electrical stimulation for ejaculation. AB - The purpose of this collective review is to discuss the technique and effectiveness of rectal probe electroejaculation in conjunction with various assistive reproductive modalities. The electroejaculation probe is positioned inside the anal ring with the electrodes placed against the prostate gland and seminal vesicles, after which electrostimulation is begun. Rectal probe electroejaculation must be always employed in a hospital setting to detect and prevent autonomic dysreflexia. This technology is considered the best approach in patients with spinal cord injury levels below T10 or in other patients in which penile vibratory stimulation fails. PMID- 12545946 TI - Experts want TIAs redefined. Transient ischemic attack symptoms are to a stroke what chest pain is to a heart attack. PMID- 12545947 TI - Mercury and your heart. Does mercury in fish increase your risk of heart disease? PMID- 12545948 TI - Alcohol's "darker side." A drink a day may raise a woman's risk of breast cancer. PMID- 12545949 TI - When tiny glands cause big problems. Kidney stones or osteoporosis may signal hyperparathyroidism, a disease that disrupts the distribution of calcium in the body. PMID- 12545950 TI - Walk this way. PMID- 12545951 TI - Pain relievers may boost blood pressure. PMID- 12545952 TI - Coping with cancer. PMID- 12545953 TI - Alternative menopause treatments fall short. PMID- 12545954 TI - Multifocal eyeglasses raise risk of falls. PMID- 12545955 TI - New cervical cancer screening guidelines. PMID- 12545956 TI - Are acupressure wrist bands helpful for motion sickness? PMID- 12545957 TI - Are there any studies showing whether ginkgo biloba is effective for tinnitus (ringing in the ears)? PMID- 12545958 TI - "Report card": a B- for cardiac care. Many hospitals could be doing better at meeting treatment guidelines. PMID- 12545959 TI - A defibrillator for home use is safe, simple--and not for everyone. PMID- 12545960 TI - Sharpening the power to predict heart attacks. PMID- 12545961 TI - Longevity's secret? It helps if mom or dad lived a century. PMID- 12545962 TI - Statins safe for elderly patients. New findings offer reassurance about the cholesterol drugs. PMID- 12545963 TI - Control hypertension to protect your memory. Keeping your blood pressure low may guard against Alzheimer's, new research suggests. PMID- 12545964 TI - Building blood vessels in the lab from skin cells. Tissue engineering may offer bypass patients a new source of "spare parts" for the heart. PMID- 12545965 TI - Ask the doctors. I was always told that low-fat diets are most appropriate for patients with heart disease. Now I'm hearing that high-fat diets are the way to go. What should I believe? PMID- 12545966 TI - Ask the doctors. I quit smoking years ago to decrease my risk of heart attacks and lung cancer. But most of the guys where I work still smoke. How much of a risk is this? PMID- 12545967 TI - Ask the doctors. Is it a good idea to take folate if you have had an angioplasty? PMID- 12545968 TI - [Syndrome differentiation and typing of traditional Chinese medicine and the clinical efficacy in 148 cases of acne vulgaris]. AB - To differentiate and type the acne vulgaris patients and to treat them with our own recipes for dissipating heat and detoxifying in traditional Chinese medicine 148 cases were typed and trated as: carbuncles and stasis (84 cases), carbuncles and damp 'toxins' (16) and damp 'toxins' and stasis (48) as well as the contrast groups of 60 patients that treated with chemical medicines. The results were: cured 86 cases (58%); significantly effective 38 cases (26%); effective 18 cases (12%); and ineffective 6 cases (4%). The contrast groups had the results of: cured 10 (17%), significantly effective 14(23%), effective 21(35%) and ineffective 15(25%). The total effective rates of both groups were 96 and 75% (P < 0.01). Following the principles of traditional Chinese medicine in differentiating and typing the acne vulgaris patients and treating them accordingly with recipes of dissipating heat and detoxifying, dissipating heat and drying the dram, eliminating the stagnant and detoxifying is an effective treatment for clinic application. PMID- 12545969 TI - [Inhibitory Effects of Etoposide Combined with 8-methoxypsoral en on Highly Metastatic Human Mucoepidermoid Carcinoma Cells]. AB - OBJECTIVE: To study the inhibitory effects of Etoposide (VP16) combined with 8 methoxypsoralen (8-MOP) on human highly metastatic mucoepidermoid carcinoma cells (Mc3). METHODS: Mc3 cells were exposed to the drugs and their combinations at various concentrations. The inhibitory effects were tested with microculture tetrazolium colorimetric assay (MTT assay) and cologenic assay. RESULTS: The IC50 values of VP16 and 8-MOP were 1625. 25 ng/ml and 750000 ng/ml respectively, the CI50 of the combination at 100320 and 1000 ng/ml of VP16 with 8-MOP were 0.350, 0.599 and 0.880 respectively. CONCLUSION: The data indicate that combination of VP16 and 8-MOP is synergic in the inhibition of Mc3 cell growth. PMID- 12545970 TI - Laboratory evaluation of Vectobac as against Aedes aegypti in Monterrey, Nuevo Leon, Mexico. AB - Intensive use of the organophosphate insecticide malathion against adults and temephos against larvae of Aedes aegypti in Mexico over the past 30 years has led to problems requiring the use of new larvicides. Toward this objective, Bacillus thuringiensis var. israelensis (Bti), a target-specific and environmentally safer control agent, was evaluated. Laboratory bioassays were done to determine the susceptibility of 2nd- and 3rd-stage larvae of Ae. aegypti to Vectobac 12 AS (aqueous suspension, 600 ITU/mg). A median lethal concentration of 0.0104 ppm and a 95% lethal concentration of 0.18 ppm were determined after 24 h of exposure to the agent. The values obtained were adjusted for field application and were further tested in the field by the State of Nuevo Leon, Mexico Vector Control Program. Suspensions of Bti were poured into pipe-water trucks and transferred to domestic 200-gal metal water drums. Larval populations were reduced during a 2 week study period. However, residents complained about a fine dusty film on the water surface. Nevertheless, these results are promising for future Bti field applications. PMID- 12545972 TI - Phase-object detection by use of double-exposure fringe-modulated speckle patterns. AB - We study the properties of a double-exposure image specklegram of a diffuse object obtained by use of a double-aperture pupil. A phase object is placed in front of one aperture during the first or the second exposure. Also, it is assumed that a uniform displacement of the diffuser between exposures is produced. The recorded specklegram is coherently illuminated and analyzed by Fourier transform operations. The average intensity distribution and the interference fringe visibility in the Fourier plane are investigated. On this basis, an alternative interference technique to detect phase objects is proposed. PMID- 12545971 TI - Pericardiocentesis and the use of enalapril in a Fischer's lovebird (Agapornis fischeri). PMID- 12545973 TI - We hardly know those eyes. PMID- 12545974 TI - Programmes for the children of illicit drug-using parents: issues and dilemmas. AB - Concern about the health and general well-being of children whose parents use illicit drugs has contributed to an increased interest in intervention programmes, but the number of such services is still limited. We review published papers about residential, home-visiting and non-residential programmes and use these and our experience in studying playgroup-based clinics to outline issues and dilemmas they face. These include balancing trust and acceptance with intervention when problems are identified, harmonizing accessibility and flexibility with the provision of child-focused activities and adult education, finding a location that is both suitable and affordable, appropriately supporting staff, collaborating with other services and securing adequate funding, including for ongoing evaluation and monitoring. PMID- 12545975 TI - VEGF induced hyperpermeability in bovine aortic endothelial cell and inhibitory effect of salvianolic acid B. AB - AIM: To examine the effect of recombinant human vascular endothelial growth factor (VEGF) on the low density lipoprotein (LDL) permeability of bovine aortic endothelial cells (BAEC) and the inhibitory effect of salvianolic acid B in vitro. METHODS: The confluence BAEC monolayers were cultured with normal medium and with medium containing VEGF or salvianolic acid B at various concentrations and for various time periods. The iodine labeled LDL flux across the monolayers was then performed, and radioactivity was measured by SN-695 automatic liquid scintillation counter. RESULTS: Addition of purified human recombinant VEGF to the BAEC monolayers could significantly increase the permeability of the monolayer to 125I-LDL (P < 0.01). The permeability-increasing activity of VEGF on the BAEC monolayers was both dose and time dependent. Salvianolic acid B could markedly inhibit the VEGF-induced hyperpermeability in BAECs (P < 0.01). CONCLUSION: VEGF plays a role in the formation and development of atherosclerosis, and salvianolic acid B has inhibitory effect on VEGF-induced hyperpermeability in BAEC. PMID- 12545976 TI - Advance directives: an overview of changing right-to-die laws. PMID- 12545977 TI - The relationship between toleration and entitlement in health care policy. PMID- 12545978 TI - Umbilical cord blood transplants: legal considerations. PMID- 12545979 TI - Virtue ethics for nurses and health care. PMID- 12545980 TI - Involuntary detention of TB patients: an age-old problem in a modern context. PMID- 12545981 TI - Akrasia, AIDS, and virtue ethics. PMID- 12545982 TI - The nurse and patient: is it a fiduciary-advocacy relationship? PMID- 12545983 TI - Ethical relationships among nurses. PMID- 12545984 TI - Wives' struggle in living through treatment decisions for husbands with advanced Alzheimer's disease. PMID- 12545985 TI - Ethics and the rural nurse: a research study of problems, values, and needs. PMID- 12545986 TI - JCAHO adopts 2000 edition of Life Safety Code. PMID- 12545987 TI - Survey due dates advanced or extended for corporate surveys. PMID- 12545989 TI - Proposed: a European cancer research area. PMID- 12545991 TI - The nurse's ethical obligation to relieve pain: actualizing the moral mandate. PMID- 12545990 TI - Issues impacting health care in the new millennium: privacy, electronic medical records, and the internet. PMID- 12545992 TI - Ethical decision making for emergency nurses: a descriptive model. PMID- 12545993 TI - An interview with Nigel Gray. PMID- 12545994 TI - The conjoined twins. Transcript of the speeches given at the BAFS annual dinner on 28 February 2002. British Academy of Forensic Sciences. PMID- 12545995 TI - The conjoined twins. Transcript of the speeches given at the BAFS annual dinner on 28 February 2002. British Academy of Forensic Sciences. PMID- 12545996 TI - Medicare and Medicaid programs; hospital conditions of participation: quality assessment and performance improvement. Final rule. AB - This final rule requires hospitals to develop and maintain a quality assessment and performance improvement (QAPI) program. In the December 19, 1997 Federal Register, we published a proposed rule to revise the hospitals conditions of participation (CoPs). The QAPI CoP was one of the conditions included in the proposed rule. We separated the QAPI CoP from the larger set of hospital CoPs so that it could be published in advance of the remaining CoPs to implement the Administration's initiatives regarding medical errors. QAPI focuses provider efforts on the actual care delivered to patients, the performance of the hospital as an organization, and the impact of treatment furnished by the hospital on the health status of its patients. Specifically, it is important to note that a QAPI is not designed to measure a hospital's quality, but rather a minimum requirement that the hospital systematically examine its quality and implement specific improvement projects on an ongoing basis. State agencies (SAs) during their surveys, review all aspects of a hospital's operations and this review provides a framework in which the SA can assess a hospital's QAPI program. In addition, the QAPI entails all activities required for measuring quality of care and maintaining it at acceptable levels. This typically includes; 1) Identifying and verifying quality-related problems and their underlying cause; 2) Designing and implementing corrective action activities to address deficiencies; 3) Following up to determine the degree of success of an intervention and to detect new problems and opportunities for improvement. Performance improvement activities aim to improve overall performance assuming that there is no permanent threshold for good performance. Under performance improvement framework, hospitals will continuously study and improve the processes of healthcare and delivery of service. PMID- 12545997 TI - Medicaid program; external quality review of Medicaid managed care organizations. Final rule. AB - This final rule establishes requirements and procedures for external quality review (EQR) of Medicaid managed care organizations (MCOs) and prepaid inpatient health plans (PIHPs). It defines who qualifies to conduct EQR and what activities can be conducted as part of EQR. In addition, under certain circumstances, this rule allows State agencies to (1) use findings from particular Medicare or private accreditation review activities to avoid duplicating review activities, or (2) exempt certain Medicare MCOs and PIHPs from all EQR requirements. Also, this rule allows the payment of enhanced Federal financial participation (FFP) at the 75 percent rate for the administrative costs of EQRs or EQR activities that are conducted by approved entities. PMID- 12545998 TI - Medicare, Medicaid, and CLIA programs; laboratory requirements relating to quality systems and certain personnel qualifications. Final rule. AB - This final rule revises and responds to comments on certain laboratory requirements issued pursuant to the Clinical Laboratory Improvement Amendments of 1988 (CLIA), Pub. L. 100-578. Specifically, this final rule sets forth requirements for certain quality control (QC) provisions and personnel qualifications; consolidates and reorganizes the requirements for patient test management, QC, and quality assurance; and changes the consensus required for grading proficiency testing challenges. To ensure a smooth transition to the new provisions for directors of high complexity testing who are not board certified (but who have doctoral degrees), we will not be holding facilities out of compliance with the provisions of the rule concerning directors who are not board certified until the effective date of this new rule, to the extent the facilities are otherwise in compliance with the requirements for laboratory directors. PMID- 12545999 TI - Managing uncontrolled hemorrhage in trauma and surgery: a novel and promising approach. PMID- 12546000 TI - Recombinant activated factor VII: its mechanism of action and role in the control of hemorrhage. AB - PURPOSE: Recombinant activated factor VII (rFVIIa) has proven both safe and efficacious in the treatment of bleeding episodes in patients with hemophilia A or B who have developed inhibitors. More recently, a growing number of reports suggests that rFVIIa may also have indications for the treatment of bleeding in patients with other hemostatic disorders, including qualitative and quantitative platelet defects, factor deficiencies other than hemophilia, and in otherwise healthy patients with uncontrollable hemorrhage following surgery or trauma. We have attempted to reconcile the various proposed mechanisms of action of rFVIIa with its apparent efficacy in such diverse clinical settings. SOURCE: A review of the literature was performed to determine those clinical scenarios in which rFVIIa appears to have been effective in controlling associated hemorrhage. PRINCIPAL FINDINGS: Findings from our group and others have demonstrated that rFVIIa is able to directly activate factor X and increase thrombin production on the surface of activated platelets in the absence of factor VIII or IX, as well as to improve thrombin generation in thrombocytopenia, and to yield a fibrin dot more resistant to fibrinolysis in vitro. CONCLUSIONS: Through these primary mechanisms, we believe that rFVIIa may be able to compensate for a variety of defects in hemostasis and merits further investigation as a general therapeutic for uncontrollable hemorrhage. PMID- 12546001 TI - Spain's Earth scientists and the oil spill. PMID- 12546002 TI - Revisiting coauthor responsibility. PMID- 12546003 TI - Possible role of recombinant activated factor VII (rFVIIa) in the control of hemorrhage associated with massive trauma. AB - PURPOSE: To report our experience with recombinant activated factor VII (rFVIIa) to control hemorrhage in trauma patients with profound multifactorial coagulopathy. rFVIIa forms a complex with tissue factor exposed at sites of tissue damage and induces activation of coagulation limited to the site of injury. It is approved for use in hemophilia patients, however, its use in trauma is still controversial due to the theoretical risk of thromboembolic complications. CLINICAL FEATURES: Nineteen critically ill, multi-transfused patents with trauma (ten blunt and nine penetrating), aged 25 + 17 yr,were treated with rFVIIa after all conventional hemostatic measures had failed. After one to three doses of rFVIIa, hemorrhaging ceased within minutes in 15/19 (78.9%) patients. The total dose of rFVIIa required to control bleeding was 195 +/- 112.7 microg x kg(-1). Shortening of prothrombin time and partial thromboplastin time was observed within 15-30 min from 22.7 +/- 7.9 to 10.4 +/- 2.6 sec and 71 +/- 38.9 to 42.2 +/- 24 sec respectively, (P < 0.05). Transfusion requirements decreased from 30 +/- 18.3 units used within 5.6 +/- 3.4 hr of admission to 2.8 +/- 2.5 within the following 24 hr (P < 0.05). One patient developed clinical deep vein thrombosis. No systemic activation of coagulation was observed clinically. Thirteen patients (68.4%) survived and recovered. Four patents did not respond to rFVIIa treatment and exsanguinated within 24 hr. Two patients died after one week, one from sepsis and one from multi-organ failure. CONCLUSIONS: rFVIIa is a promising adjunctive hemostatic treatment for trauma patients suffering from massive bleeding. Controlled trials are warranted to evaluate the safety and efficacy of this drug. PMID- 12546004 TI - The next generation of science policy-makers. PMID- 12546005 TI - Heat and biodiversity. PMID- 12546006 TI - Gluten peptides and celiac disease. PMID- 12546007 TI - A novel hemostatic agent: the potential role of recombinant activated factor VII (rFVIIa) in anesthetic practice. AB - PURPOSE: To review the role of recombinant factor VIIa in anesthetic practice. SOURCE: A review of the published literature. MAIN FINDINGS: The mechanism of action of rFVIIa suggests enhancement of hemostasis limited to the site of injury without systemic activation of the coagulation cascade. In addition to its indication for use in patients with hemophilia, use of rFVIIa for treatment of uncontrolled massive hemorrhage in various peroperative settings appears to be rational, safe, and effective. Published results suggest that in trauma patients rFVIIa may play a role as an adjunctive hemostatic measure in addition to surgical hemostatic techniques There is preliminary evidence that hemorrhagic complications (eg. epistaxis, vaginal bleeding) associated with profound thrombocytopenia can be reversed with rFVIIa even at platelet counts below 10,000 per microL. Various case reports outlining the successful treatment with recombinant factor VIIa of patents experiencing intractable bleeding after valve replacement surgery, and with severe hemorrhage during therapy with left ventricular assist device, indicate the potential therapeutic efficacy of this agent in cardiac surgical procedures. Additionally, rFVIIa has been used successfully for treatment of massive postoperative bleeding following general surgery. CONCLUSIONS: rFVIIa is a novel hemostatic agent that shows promise in non-hemophiliac patents of a significant therapeutic role in variety of coagulopathic and hemorrhagic conditions in clinical situations ranging from thrombocytopenia, disseminated intravascular coagulation and transfusion-related coagulopathy, as well as in patients experiencing massive blood loss undergoing orthotopic liver transplantation, cardiac, orthopedic and genitourinary surgery. PMID- 12546008 TI - Restaging after induction therapy for non-small cell lung cancer. PMID- 12546009 TI - IRB triage of projects that involve medical record review. PMID- 12546010 TI - Ethical issues arising when interim data in clinical trials is restricted to independent data monitoring committees. PMID- 12546011 TI - What medical school did you graduate from? Judicial prescribing of medical treatments. AB - There is much in the literature concerning conflicts between clinicians and the law over who should make decisions in a legal context, such as involuntary hospitalization or patients' competency to make treatment decisions. There is little, however, about judges' trying to impose specific treatment decisions on clinicians. This article addresses that situation and the problems such orders cause. PMID- 12546012 TI - When religion may be an ethical issue. AB - Though Americans are a religious people, there are times when religion or religious views may confound the ethical process. This article claims that religious values may be expressed as either principles or narratives, then seeks to establish a case for dealing with religious claims as "principled narratives." Methods of evaluation are explored and then seven guidelines are offered for dealing with religion in ethics consultations. PMID- 12546013 TI - Catholic health care cooperation: why rewrite the ethical and religious directives? PMID- 12546014 TI - Revising the ERDS'94: goals, opposition and resolution. PMID- 12546015 TI - Direct sterilization: an intrinsically evil act--a rejoinder to Fr. Keenan. PMID- 12546016 TI - Needle exchange programs and HIV prevention: the greater good or the promotion of evil? PMID- 12546017 TI - A law school forum on human cell-lines and frozen embryos. PMID- 12546018 TI - Eight simple rules for starting that new job. PMID- 12546019 TI - "We'll leave the light on.". PMID- 12546020 TI - Ethical implications of research results--what is to be done with the results gathered under the Nazi regime? PMID- 12546021 TI - Cryptogam blue-light photoreceptors. PMID- 12546022 TI - How hemolysis causes acute pancreatitis. PMID- 12546023 TI - Does hyperhomocysteinemia due to vitamin B12 deficiency associted with Helicobacter pylori infection has a role on cerebral stroke? PMID- 12546024 TI - Insurance repercussions of mammographic screening: what do women think? PMID- 12546025 TI - TGF-beta1 and collagen methabolism in IBD patients. PMID- 12546026 TI - More over anthrax. PMID- 12546027 TI - Home visits by paraprofessionals did not improve maternal and child health. PMID- 12546028 TI - A behavioural infant sleep intervention resolved sleep problems. PMID- 12546029 TI - Review: extended stimulant medication is effective in children with attention deficit hyperactivity disorder. PMID- 12546030 TI - Review: "Scared Straight" and other juvenile awareness programmes increase criminal behaviour. PMID- 12546031 TI - Review: primary prevention strategies do not delay initiation of intercourse, improve contraceptive use, or reduce pregnancies in adolescent women. PMID- 12546032 TI - Mifepristone was more effective than the Yuzpe regimen for emergency contraception. PMID- 12546033 TI - Review: antibiotic treatment for 7-14 days reduces treatment failure in children with urinary tract infection. PMID- 12546034 TI - Review: probiotics are effective in preventing antibiotic associated diarrhoea. PMID- 12546035 TI - Treatment of scalp lacerations with a hair apposition technique reduced scarring, pain, and procedure duration compared with suturing. PMID- 12546036 TI - Review: chlorhexidine gluconate is more effective than povidone-iodine for preventing vascular catheter related bloodstream infection. PMID- 12546037 TI - Review: organisational change and patient directed strategies may increase adult immunisation and cancer screening. PMID- 12546038 TI - Oestrogen plus progestin increased coronary heart disease and breast cancer in postmenopausal women. PMID- 12546040 TI - Doppler echocardiography was not accurate for diagnosing asymptomatic thrombosis associated with an umbilical venous catheter in infants. PMID- 12546039 TI - Nurse led transitional care improved health related quality of life and reduced emergency department use for heart failure. PMID- 12546041 TI - The CRAFFT test was accurate for screening for substance abuse among adolescent clinic patients. PMID- 12546042 TI - Review: specific signs and symptoms can help practitioners to diagnose acute purulent sinusitis in general practice. PMID- 12546043 TI - Functional neurological deficit increased the risk of death in childhood epilepsy. PMID- 12546044 TI - Review: some preschool children develop complications after presenting to primary care with acute cough. PMID- 12546045 TI - Evidence based patient information leaflets in maternity care had limited visibility and did not promote informed choice of childbearing women. PMID- 12546046 TI - Mothers progressed through a 4 phase process to resume their own lives after having twins. PMID- 12546047 TI - Review: mothers of children with physical or mental disabilities experience emotional compromise between acceptance and denial. PMID- 12546048 TI - Women with polycystic ovarian syndrome had excess hair, irregular or absent menstruation, and infertility and felt freakish, abnormal, and not proper women. PMID- 12546049 TI - Active consideration underpinned the efforts of men with prostate cancer to support their spouse caregivers. PMID- 12546050 TI - The lived experience of men with erectile dysfunction was reflected in the themes of loss and being alone with it. PMID- 12546051 TI - Epidemiology of rhinitis in Thais: characteristics and risk factors. AB - A questionnaire survey was performed on 3,124 subjects living in Bangkok and its vicinity. The cumulative prevalence of chronic rhinitis (CR) was 13.15% (95% CI = 13.13-13.17). The characteristics of CR subjects, i.e. age group, current occupation, associated allergy, family history of atopy, cigarette smoking and drinking habits were totally different from the non CR group, except for sex preponderance. The possible risk factors for developing CR in this group were high income occupation, presence of associated allergy, family history of atopy, smoking and drinking habits. When CR subjects were further classified into "allergic" and "non-allergic" CR groups by using the presence of associated eye symptoms and known provoking factors as the differential criteria, there was no significant difference in all parameters compared between the two groups, i.e. age, sex, current occupation, associated allergy, family history of atopy, smoking and drinking habits, frequency of occurrence of each nasal symptom, seasonal variation of the symptoms, effect of moving to live in the big city, presence of pets in the house and effect of changing occupation, except for the number of nasal symptoms which was significantly higher in the "allergic" CR group. PMID- 12546052 TI - Isolation and characterization of allergens from the seeds of Vigna sinensis. AB - Allergenic components of cowpea vegetable green seeds (Vigna sinensis) were isolated based on solubility, isoelectric precipitation and molecular mass. The allergenicity of the cowpea fractions was monitored by enzyme-linked immunosorbent assay (ELISA) and skin-prick test. The allergenic albumin fraction was characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and IgE-specific immunoblotting. The 41 and 55 kDa protein components were found to be major allergens and the allergenicity was resistant to heat and proteolytic enzyme digestion. This study confirms the presence of potent allergens in cowpea seeds. PMID- 12546053 TI - Prevalence of wheeze, night cough and doctor-diagnosed asthma in pre-school children in Kota Bharu. AB - While many studies of the prevalence of wheeze have been conducted in schoolchildren, there have been few in pre-school children. Most children with asthma develop symptoms before the age of 5 years and many pre-school wheezers continue to wheeze in the early school years. Among the latter, those children who continue to wheeze at school age have poorer lung function than those who don't. It is thus appropriate to enquire more fully about wheeze in this age group where its incidence is high and its relation with asthma less well defined. The objective of this study was to investigate the prevalences of wheeze, night cough and doctor diagnosed asthma in pre-school children. A cross-sectional study was conducted in five primary health clinics in the district of Kota Bharu from April to October 1998. Nurses from these clinics distributed Bahasa Malaysia questionnaires containing questions on asthma symptoms to preschool children aged 1-5 years during their home visits. The respondents were parent(s) or carer(s) of the child. The response rate was 100% and a total of 2,878 responses were analysed. The prevalence of symptoms and doctor diagnosed asthma were as follows: ever wheezed 9.4% (95% confidence interval (CI) 8.3-10.4%); current wheeze 6.2% (95% CI 5.2 to 7.0%); night cough 10.2% (95% CI 9.1 to 11.4%); and doctor diagnosed asthma 7.1% (95% CI 6.2 to 8.0%). There were no significant differences in prevalence between males and females, or among age groups. The prevalence of night cough in children with no history of wheeze was 6.9%. The cumulative and current prevalences of wheeze were similar to, and those of night cough and doctor-diagnosed asthma significantly lower than, those reported for Kelantan schoolchildren. These findings provide a baseline for assessing future symptoms trends, and perhaps also the validity of diagnosing asthma in this age group. PMID- 12546054 TI - Reticulocyte analysis in systemic lupus erythematosus and chronic renal failure using flow cytometry. AB - The number and maturation of circulating reticulocytes were measured in patients with systemic lupus erythematosus (SLE) and chronic renal failure (CRF) using an automated hematological analyzer (Technicon H*3 RTX) for their erythropoietic activities. Both SLE and CRF patients had increased reticulocyte numbers with a low degree of maturation. The SLE patients had no changes in mean reticulocyte corpuscular volume (MCVr) as compared to normal subjects (110.20 +/- 15.43 fl. in SLE and 110.39 +/- 5.09 fl. in normal), whereas CRF patients had significantly increased mean corpuscular reticulocyte volume (MCVr = 120.99 +/- 8.09 fl., p value = 0.0019 as compared with normal). Three cases of SLE with nephrotic syndrome (NS) had high degree of MCVr (113.4, 125.0 and 133.1 fl., respectively). The renal involvement in SLE patients and CRF patients may associate with increased reticulocyte corpuscular volume. PMID- 12546055 TI - Rosetting of Plasmodium falciparum required multiple components of the uninfected erythrocytes. AB - The mechanism of rosette formation of uninfected erythrocytes with Plasmodium falciparum-infected erythrocytes is rarely described. In this study, rosetting of uninfected normal erythrocytes with infected erythrocytes significantly reduced after treatment of the uninfected erythrocytes with neuraminidase. In contrast, the rosetting property of the infected erythrocytes was abolished by trypsinization but not by neuraminidase. The in vitro rosetting model showed that uninfected thalassemic erythrocytes poorly formed rosettes with infected normal erythrocytes when compared with normal erythrocytes of the same blood group. A rosetting parasite clone showed significant reduction in rosetting with thalassemic erythrocytes of all blood groups, however, this reduction was not obvious when the wild P. falciparum isolates were studied. These results suggest that while parasites from a single clone can rosette with uninfected erythrocytes via carbohydrate component, there is more than one type of receptor on uninfected erythrocytes involved in rosette formation with the heterogeneous populations of the wild P. falciparum isolates. PMID- 12546056 TI - Trichinella spiralis-specific monoclonal antibodies and affinity-purified antigen based diagnosis. AB - Hybridomas secreting monoclonal antibodies (MAbs) to Trichinella spiralis were produced. Myeloma cells were fused with splenocytes of a mouse immunized with excretory-secretory (E-S) antigen of infective larvae. A large percentage of growing hybrids secreted antibodies cross-reactive to many of 23 heterologous parasites tested. Only 6 monoclones (designated 3F2, 5D1, 10F6, 11E4, 13D6 and 14D11) secreted MAbs specific to the E-S antigen and/or a crude extract (CE) of T. spiralis infective larvae. The 6 monoclones secreted IgM, IgG3, IgM, IgG3, IgG3 and IgG3, respectively. Clone 5D1 was selected to mass produce MAbs which were then coupled to CNBr-activated Sepharose CL-4B to prepare an affinity purified antigen. Dot-blot ELISA with either purified antigen or CE was evaluated. There were 17 patients with acute trichinellosis and 76 individuals convalescing from T. spiralis infection (group 1). Controls were 170 patients with parasitic infections other than trichinellosis (group 2) and 35 healthy parasite-free controls (group 3). CE-ELISA was positive in all group 1 patients. However, sera from many group 2 patients also were reactive (opisthorchiasis 44.2%, schistosomiasis-44%, gnathostomiasis-30%, paragonimiasis-28.6%, taeniasis 27.3%, strongyloidiasis-23.1% and hookworm infections-20%). Affinity-purified antigen was 100% specific, all sera from group 2 and group 3 individuals tested negative. Although 74 of 76 patients (97.4%) with convalescing trichinellosis tested positive, sera from only 3 of 17 patients (17.6%) with acute T. spiralis were reactive. Thus, CE antigen is appropriate when sensitivity is needed, while purified antigen should be used when specificity is required. Dot-blot ELISA is easier to perform, more rapid and less expensive than indirect ELISA. Many samples can be assayed simultaneously, special equipment is not required, and results can be preserved for retrospective analysis. Dot-blot ELISA is therefore the method of choice for the rapid diagnosis of trichinellosis, particularly when more complex laboratory tests are unavailable. PMID- 12546057 TI - Antigenic components of Gnathostoma spinigerum recognized by infected human sera by two-dimensional polyacrylamide gel electrophoresis and immunoblotting. AB - Antigenic components of Gnathostoma spinigerum larval extract were revealed by two-dimensional gel electrophoresis (2-DE) and immunoblot analysis using sera from patients with 6 proven cases of gnathostomiasis, 5 presumptive cases of gnathostomiasis, 3 proven cases of angiostrongyliasis, 3 proven cases of cysticercosis, and pooled sera from healthy adults. By the 2-DE, the larval extract was highly complex and consisted of more than 75 polypeptides. Immunoblotting analysis of this larval extract after reaction with each of 6 proven gnathostomiasis sera revealed various numbers of antigenic spots ranging from 30 to 70 spots at the approximate molecular masses of less than 14.4 to more than 94 kDa with isoelectric points (pI) of less than 4.65 to 9.6. Antigenic spots at the approximate molecular mass of more than 30 kDa were recognized with the proven angiostrongyliasis, proven cysticercosis and healthy control sera but these sera did not react with the spots at approximate molecular masses of 23-25 kDa with pI of 8.3-8.5. The reacted spots, which consisted of at least 1 to 2 spots, were unique for the recognition of gnathostomiasis sera. Five out of 6 (83.3%) proven and 4 out of 5 (80%) presumptive gnathostomiasis sera reacted with these specific spots. PMID- 12546058 TI - Production of anti-CD14 monoclonal antibodies using CD14 expressing COS cells as immunizing antigen. AB - CD14 is a leukocyte surface molecule expressed on monocytes but not on lymphocytes. Recently, CD14 molecule was demonstrated to function as a receptor for endotoxin. CD14 specific monoclonal antibody (MAb), therefore, can be used to identify monocytes and study the host defense mechanism to bacterial endotoxin. To produce MAb against CD14 protein, in this study cDNA encoding CD14 protein and COS cell expression systems were used to prepare CD14 expressing COS cells. The CD14 transfectants were then used as antigen for mouse immunization. The spleen cells of the immunized mouse were then fused with myeloma cells by conventional hybridoma technique. By using this strategy, 5 hybridroma clones secreting antibody specific for CD14 molecule were generated within one fusion. The generated CD14 MAbs were strongly positive with monocytes, weakly positive with neutrophils but negative with lymphocytes. In addition, the generated CD14 MAb blocked the binding of lipopolysaccharide (LPS) to the CD14 molecules. These CD14 MAbs could be used to enumerate peripheral blood monocytes as well as using referent CD14 MAb. We, therefore, introduce an alternative method for preparation of antigen for production of monoclonal antibody. This type of antigen is a very effective antigen for the production of monoclonal antibodies against cell surface molecules. PMID- 12546059 TI - Serodiagnosis of Helicobacter pylori infection by immunoblot assay. AB - We compared a noninvasive serological test using a commercial immunoblot assay (Helicoblot 2.0) to tissue-based methods [rapid urease test (CLO test), histology and culture] in eighty Thai patients undergoing upper endoscopy. A true positive test was defined as at least two of the biopsy-related tests being positive. The CLO test was the most accurate test with sensitivity and specificity as high as 100%, whereas histology and culture had sensitivity of 100% and 72.2%, respectively, and the specificity of 72.7% and 96%, respectively. The serological test had a high sensitivity (97.2%) but exhibited an unsatisfactory specificity (40.9%). We concluded that the rapid urease test using multiple gastric biopsies was the most appropriate method for diagnosing H. pylori status. The role of immunoblot assay as a serological screening test in our population remains doubtful, but it may identify patients who have been infected with certain strains of H. pylori. PMID- 12546060 TI - Atopic cataracts in a child with atopic dermatitis: a case report and review of the literature. AB - Cataracts induced by atopic dermatitis rarely occur in adolescent and young adult patients suffering from this problem. Lenticular opacity is an important ocular complication in atopic dermatitis. Although the cause of atopic dermatitis and its ocular complications are unknown, cataracts have been observed to develop and progress during periods of exacerbation of the dermatitis. We report the case of a 16-year-old boy with atopic dermatitis who abruptly developed cataracts in both eyes while suffering from severe skin itching which began 2 months before the initial examination. His peroxidation test result was very high, and we postulate the retinal peroxidation might play a key role in cataractogenesis. Lens aspiration and intraocular artificial lens implantation were performed smoothly with restoration of visual acuity in both eyes. PMID- 12546062 TI - Ludwik Hirszfeld: a life in serology. PMID- 12546061 TI - Cloning and expression of a novel neurotrophin, NT-7, from carp. AB - Neurotrophins have been demonstrated to play important roles in the development and functioning of the nervous system. This family of proteins consists of four homologous members in mammals: NGF, BDNF, NT-3, and NT-4/5. A new member, called NT-6, was recently cloned from the platyfish Xiphophorus maculatus. This protein shares closer structural relationship to NGF than the other neurotrophins, but contains a characteristic insertion of 22 amino acids that constituted the heparin-binding domain. Here we report the cloning of a novel neurotrophin from the fish Cyprinus carpio (carp), which shared about 66% amino acid identity to Xiphophorus NGF and NT-6. The neurotrophin, designated NT-7, possesses structural characteristics common to all known neurotrophins, such as the presence of six conserved cysteine residues and the flanking conserved sequences. In addition, there is an insertion of 15 amino acids at the position corresponding to that observed for NT-6. The neurotrophic activity of NT-7 was demonstrated by its ability to promote neurite outgrowth and neuronal survival of chick dorsal root ganglia. Phosphorylation assay of various Trk receptors overexpressed in fibroblasts suggested that NT-7 could activate TrkA but not TrkB or TrkC. Northern blot analysis revealed that NT-7 was predominantly expressed in peripheral tissues, though weak expression was also detected in the brain. Like NT-6, this novel neurotrophin might represent yet another NGF-like neurotrophin in lower vertebrates. PMID- 12546063 TI - Predicting outcome in hematological stem cell transplantation. AB - This review summarizes recent results investigating the role of certain cytokine gene polymorphisms, including those of TNF-alpha, IFN-gamma, IL-6, IL-10 and IL-1 receptor antagonist (IL-1Ra), in allogeneic stem cell transplantation. It discusses their role in predicting outcome and the development of a genetic risk index for graft versus host disease (GvHD) in HLA-matched sibling transplants. By the comparative use of an in vitro human skin explant model, initial results suggest that certain cytokine gene polymorphisms may be associated with more severe disease. PMID- 12546064 TI - Structures and serology of the O-specific polysaccharides of bacteria of the genus Citrobacter. AB - The review presents the structures of the O-specific polysaccharides (O-antigens) of the lipopolysaccharides isolated from over 25 Citrobacter strains, which represent different species and serogroups. The correlation between O-antigen structure and immunospecificity as well as numerous cross-reactions between Citrobacter and other enterobacterial species are discussed. PMID- 12546065 TI - Streptomyces and Escherichia coli, model organisms for the analysis of the initiation of bacterial chromosome replication. AB - Streptomyces coelicolor A3(2) and Escherichia coli are quite different in their life-style and the structures of their genomes. Streptomyces exhibit complex multicellular development including formation of multigenomic hyphae during growth. These organisms possess a large linear (8.7 Mb) and GC-rich (approximately 72%) chromosome. The genome sequence of S. coelicolor has just been completed. The difference between E. coli and Streptomyces making them an excellent model organisms for a comparison of their replication modes. In this review, we compare initiation of chromosome replication in both organisms. Their replication origins are different in size, but both have DnaA boxes--a binding motifs for initiator DnaA protein. The two DnaA proteins have practically the same biochemical properties. Many aspects of the control of initiation seem to be similar. A comparison of the two systems thus allows us to define those aspects of replication initiation that are universally used in the eubacterial kingdom. PMID- 12546066 TI - Lactoferrin and immunologic dissonance: clinical implications. AB - Homeostasis is the maintenance of equilibrium in a biological system by means of positive and negative feedback control mechanisms that counteract influences tending toward physiological dissonance. At the molecular level, homeostasis is controlled by the network of the neuro-endocrine-immune system, in which lactoferrin (LF) plays a central role. The purpose of this review is to provide a comprehensive summary of a collaborative study established between the Hirszfeld Institute of Immunology and Experimental Therapy (Wroclaw, Poland) and the University of Texas Health Science Center (Houston, USA) regarding LF and its role in homeostasis. In our studies we focused on the immunoregulatory functions of LF, both in vitro and in vivo. We investigated the immune status of individuals subjected to different insults, including experimental endotoxemia in mice and surgery in humans. We also studied a LF-dependent delayed type hypersensitivity (DTH) response to evaluate some of the mechanisms by which LF can effectively substitute an adjuvant in vaccine. PMID- 12546067 TI - Experimental therapies for psoriasis. AB - There is a high medical need for better therapies for psoriasis. Based on new insight into the pathophysiology of this frequent immune disease, a number of novel systemic immunomodulatory therapies are currently in clinical development. These include approaches targeting antigen presentation and costimulation, T cell activation and leukocyte adhesion, action of proinflammatory mediators, and modulating the cytokine balance. Although mainly only preliminary data are available so far, these trials contribute to a further understanding of the disease and will eventually lead to new therapeutic options for psoriasis. Moreover, since psoriasis can be considered as a visible model disease for T cell mediated disorders characterized by a type 1 cytokine pattern in general, such approaches may have impact for other immune disorders as well. Here we review the rationale and the initial clinical data of these important recent experimental therapies. PMID- 12546068 TI - Diminished expression of the type II receptor for TGFbeta (TGFbetaRII) in T lymphocytes from patients with Sezary syndrome is not due to mutations in the receptor's poly-A tract: limitations of the standard RT-PCR in cDNA sequence analysis of homopolymeric base stretches. AB - Peripheral blood lymphocytes from patients with Sezary syndrome (SzS) frequently demonstrate decreased surface expression of transforming growth factor beta receptor II (TGFbetaRII). The mechanism of this low TGFbetaRII expression remains unknown. Because mutations within the poly-A tract of the TGFbetaRII sequence (nucleotides 709-718) were shown to result in diminished TGFbetaRII expression in other types of malignant tumors, we examined the sequence of the TGFbetaRII poly A tract in two SzS-derived cell lines and in peripheral blood SzS cells from 17 SzS patients and 4 control, healthy individuals using DNA sequencing and single stranded conformation polymorphism (SSCP) analysis. A standard bidirectional, automated sequence analysis of the RT-PCR-generated cDNA TGFbetaRII fragment showed a heterogenous population of the normal length, 10-, with admixed, shortened, 9-base poly-A stretches. Surprisingly, this mixture was present not only in the cells from 5 SzS patients and 2 SzS cell lines, but also in cells from 2 healthy control individuals. Importantly, the proportion of the shortened, 9-base fragments was markedly reduced or practically eliminated when the procedure was modified by usage of high-fidelity DNA polymerase, labeled primers and/or cloned RT-PCR products, which indicates that the presence of the shortened, 9-base fragments represented a procedural phenomenon rather than a true deletional mutation within an allele of the TGFbetaRII gene. Accordingly, SSCP analysis of genomic DNA did not reveal any mutations within the poly-A tract containing region. These results indicate that a mechanism different from mutations in the polyadenine tract underlies the diminished TGFbetaRII expression in SzS cells and that the results of an unmodified, direct sequence analysis of homopolymeric base streaches in RT-PCR-derived cDNA should be interpreted with caution. PMID- 12546069 TI - Closed endarterectomy for diffuse right coronary artery disease: early results with angiographic controls. AB - BACKGROUND: Due to reported high morbidity and mortality, surgeons tend not to endarterectomize the diffusely diseased right coronary artery. Most prefer not to bypass this artery in such situation. We compared the endarterectomized right coronary patients with patients having total occlusion of right coronary artery not needing endarterectomy who received right coronary artery (RCA) bypass. MATERIAL AND METHODS: Between August 1998 and May 2000, 1226 patients had coronary artery bypass grafting (CABG) at Alkan Hospital, of whom 59 had right coronary artery endarterectomy (REC) along with RCA bypass with saphenous vein graft. We compared the results of 59 RCE patients with 50 patients who underwent RCA bypass without RCE. RESULTS: Compared with the control group, the RCE group had a higher incidence of diabetes. There were no statistically significant differences between groups for mortality and morbidity. Among postoperative variables only the cross-clamp time was longer for the RCE group (33.5 +/- 10 min vs. 27.3 +/- 8 min, p < 0.05). Surviving patients were followed for a mean period of 7 +/- 5 months. No recurrence of angina occurred during the follow-up. Control angiography at the sixth month was performed on 42 RCE patients accepting the procedure, and all grafts were found patent. CONCLUSION: Endarterectomy for diffusely atherosclerotic right coronary artery for selected patients did not bring additional mortality and morbidity to the CABG procedure, and short term clinical and angiographic results were excellent. PMID- 12546070 TI - Does coronary endarterectomy increase the risk of coronary bypass? AB - BACKGROUND: With increased incidence of angioplasty and stent implantation, patients referred for coronary bypass (CABG) typically have more advanced and diffuse coronary disease. Thus, more patients may require endarterectomy in order to achieve complete revascularization. We compared our results in patients undergoing CABG with or without coronary endarterectomy. METHODS: Between 1993 and 1999, 2372 patients underwent isolated CABG in our department. A retrospective analysis was performed to compare patients requiring coronary endarterectomy of the LAD (group 1, n = 88), endarterectomy of arteries other than the LAD (group 2, n = 143), to those not requiring endarterectomy (group 3, n = 2071). Patients undergoing CABG without the use of cardiopulmonary bypass were excluded. Group 1 had a higher incidence of proximal LAD stenosis (p = 0.001) than group 3, while group 2 had a higher incidence of peripheral vascular disease (p = 0.02), preoperative MI (p = 0.03) and LV dysfunction (p = 0.001). RESULTS: Operative mortality was 10% in group 1 (p < 0.001) and 4% in group 2 (p = NS) compared to 3% in group 3. Incidence of perioperative MI was 12% in group 1 (p = 0.001) and 8% in group 2 (p = 0.001) compared to 2% in group 3. CONCLUSIONS: Patients requiring endarterectomy of the LAD are at increased risk of operative mortality. This was not true for patients requiring endarterectomy of arteries other than the LAD. In both groups there was an increased risk of perioperative myocardial infarction. PMID- 12546071 TI - Myocardial infarction in coronary bypass surgery using on-pump, beating heart technique with pressure- and volume-controlled coronary perfusion. AB - BACKGROUND: Even with the current aerobic techniques in myocardial protection, perioperative myocardial infarction can still occur. In this study, we sought to find out whether there is a method-specific mechanism of ischemic injury in coronary bypass surgery using an on-pump beating heart technique. For this reason, we investigated localization and the extent of myocardial infarction in correlation to the severity of coronary artery stenosis. We discuss strategies for reduction of infarction incidence. METHODS: 34 patients, who after isolated coronary bypass procedure developed non-Q or Q wave infarction were selected for the study. In 12 of 34 patients (group A) beating heart technique and in 22 of 34 patients (group B) cardioplegic arrest was used for myocardial protection. The study was conducted retrospectively and included patients with stable, unstable, and postinfarction angina without preoperative enzymatic evidence of ischemic injury and with technically noncomplicated coronary bypass grafting. Excluded from the study were emergency and redo procedures. In group A, in all instances, the left anterior descending artery was grafted as the last vessel and while distal anastomoses were constructed, coronary perfusion was maintained by using a perfusion catheter. RESULTS: Most patients in both groups had triple-vessel disease (10 of 12 patients group A; 17 of 22 patients group B), all patients received complete revascularization; 7 of 12 patients in group A and 5 of 22 patients in group B had significant or critical left main stem stenosis. In group A, 11 infarctions occurred in LAD-(12 stenotic), 1 in CX-(11 stenotic), and 1 in RCA-(11 stenotic) supply area, four were Q wave infarctions. In group B, eight infarctions occurred in LAD (22 stenotic), four in CX (17 stenotic), and 14 in RCA (20 stenotic) supply area, eight were Q wave infarctions. In group A, the infarction incidence in the LAD area was 10-times higher than in CX and RCA areas. In group B, the infarction incidence in the RCA area was 2- and 3-times higher than in the LAD and CX areas, respectively. In both groups no correlation between infarction incidence and severity of stenosis was observed. CONCLUSIONS: Using an on-pump beating heart technique, higher coronary perfusion pressures, avoidance of extreme upward retraction of the heart during revascularization of the CX-branch, as well as choosing the revascularization of the LAD as the first vessel, could possibly contribute to better myocardial protection. In hearts arrested with cardioplegic solution, the right ventricle is probably more susceptible to ischemic injury, especially when RCA is poorly collateralized. For adequate protection, choosing the revascularization of the RCA as the first vessel with immediate repeated cardioplegia via a RCA graft, higher perfusion pressures and antegrade with retrograde cardioplegia delivery, may be advantageous. PMID- 12546073 TI - Valve surgery in octogenarians with a "porcelain" aorta and aortic insufficiency. AB - We report the technique of balloon occlusion of the ascending aorta in two patients requiring valve operations. In the first patient, it provided a rapid solution to unexpected and potentially catastrophic severe aortic insufficiency (AI) with the initiation of cardiopulmonary bypass in a patient who required aortic valve replacement using deep hypothermic circulatory arrest due to a "porcelain" aorta. In the second patient, it allowed for a routine mitral valve repair to be performed by providing aortic occlusion in the setting of AI and avoiding the devastating consequences of cross-clamping a "porcelain" aorta. PMID- 12546072 TI - S100B blood levels correlate with rewarming time and cerebral Doppler in pediatric open heart surgery. AB - BACKGROUND: Brain hyperthermia, accompanying the rewarming phase of cardiopulmonary bypass (CPB), has been involved in the genesis of postoperative brain damage. Blood S100B levels are emerging as a marker of brain distress, and could offer a reliable monitoring tool at different times during and after open heart surgery. METHODS: Thirty-two patients undergoing repair of congenital heart disease with CPB and deep hypothermic circulatory arrest (DHCA) were monitored by S100B blood levels and middle cerebral artery Doppler velocimetry pulsatility index (MCA PI) before, during, and after surgical procedure at five predetermined time-points. RESULTS: Both S100B and MCA PI significantly increased, MCA PI values exhibiting a peak at the end of surgery time-point (p > 0.05), while S100B blood levels were increased at the end of CPB (p < 0.05). Multivariate analysis, with S100B levels measured at the end of CPB as dependent variable, showed a positive significant correlation with MCA PI (p = 0.04), with the CPB and the rewarming duration (p = 0.03 and p = 0.009, respectively). CONCLUSIONS: The present results show a significant correlation between a biochemical marker of brain damage and an index of increased cerebrovascular resistance, with higher levels during the rewarming CPB phase in pediatric open heart surgery. PMID- 12546074 TI - A minimally invasive alternative approach for reoperative grafting of the right coronary artery. AB - BACKGROUND: Target vessel revascularization on a beating heart via a mini subxiphoid incision, not only eliminates the hazards of sternal reentry, but also avoids the detrimental systemic effects of extracorporeal circulation. AIM: The goal of the study was to develop and describe a safe and effective method to revascularize the right coronary artery in reoperative cases, using beating heart and minimally invasive techniques in lieu of the RGEA. METHODS: There were three men and four women with a mean age of 69 years; four were second time reoperations and three third time reoperations. Through a subxiphoid approach, a lower ministernotomy is performed. Once the coronary target is established, a Rultract retractor is used to gently elevate the right half of the sternum to take down the RITA. A mechanical stabilizer was used to provide stabilization for distal anastamoses. Flow was measured in all the grafts with the Medi-Stim before and after protamine administration. RESULTS: [table: see text] CONCLUSION: We believe that this technique provides another effective approach to the patient who requires reoperative coronary revascularization of the RCA. PMID- 12546075 TI - Minimally invasive surgical closure of patent ductus arteriosus in premature infants: a novel approach. AB - Recent advancement of minimally invasive cardiac surgical procedures have prompted the use of minimal incisions to perform operations on patients with congenital heart defects. In this report, we describe a new technique for closure of a patent ductus arteriosus, using an anterior extrapleural approach. We present our technique and results in five low-weight infants. This approach is safe and effective, shortens hospital stay, and is less invasive than classical approaches. PMID- 12546076 TI - Saphenous vein graft aneurysm: potential for mistaken identity. AB - We report the unusual presentation of a saphenous vein graft aneurysm in a 72 year-old man who had undergone coronary artery bypass surgery 15 years prior to admission. The patient presented with a large mass on the anterior wall of the right ventricle and chest discomfort; initially believed to be a cardiac tumor. The patient underwent reoperative coronary surgery with excision of the saphenous vein graft aneurysm, followed by an uneventful postoperative course. Aortocoronary saphenous vein graft aneurysms are rare and their presentation may be mistaken for pulmonary or chest wall lesions. Complications include rupture, and therefore require attentive diagnosis and surgical therapy. PMID- 12546077 TI - Wei Lun Visiting Professorial Lecture: Nitric oxide in the regulation of vascular function: an historical overview. AB - The field of nitric oxide (NO) research has developed in explosive proportions since the discovery of endogenous NO in 1986. The biological importance of NO was first shown by the findings that nitroglycerin causes vasodilation by liberating NO in the smooth muscle, and activating guanylate cyclase to raise smooth muscle levels of cyclic GMP. NO also inhibits platelet aggregation by cyclic GMP mechanisms. NO activates guanylate cyclase by heme dependent mechanisms involving the formation of a nitrosyl-heme complex. The high pharmacological potency of NO was finally understood when NO was shown to be formed endogenously, and to be the same as EDRF. Based on these properties of NO, new drugs can be developed as vasodilators and antiplatelet agents for the treatment of a variety of vascular disorders including impotency. NO elicits many other actions in mammalian systems including inhibition of cell proliferation, airway bronchodilation, antimicrobial effects, other host defense effects, and also modulates learning and memory as well as other central functions. This allows for an extensive opportunity to develop novel drugs for the diagnosis, prevention, and treatment of a number of different diseases, many of which are vascular in origin. PMID- 12546078 TI - Nitric oxide and fetal coronary regulation. AB - The determinants of coronary flow in the heart were studied using the chronic near-term fetal sheep model. Coronary flows were measured using implanted Doppler probes on the fetal circumflex artery calibrated with radiolabelled microspheres. Experiments were conducted to calculate maximal coronary flow under conditions of systolic work, chronic, and acute hypoxemia. Pressure-flow conductance curves were also constructed during adenosine administration. These studied showed that maximal right ventricular systolic work increases flow from a resting level of some 200 mL x min(-1) x 100 g(-1) to only about 60% of the maximal coronary flow under chemical vasodilation with adenosine (800 mL x min(-1) x 100 g(-1)). Chronic hypoxemia leads to a resting flow of some 800 mL x min(-1) x 100 g(-1) but with a remaining reserve of some 400 mL x min(-1) x 100 g(-1). Nitric oxide synthase blockade with N(u)-nitro-L-arginine (L-NNA) depresses coronary flow at all levels of oxygen content and depresses myocardial oxygen consumption even under normoxemic conditions. Fetal coronary flow increases dramatically during severe acute hypoxemia and may exceed the maximal levels found during adenosine administration without a loss of ventricular function. However, in the presence of L-NNA and severe hypoxemia, coronary flow does not exceed flows found during adenosine administration. PMID- 12546079 TI - Nitric oxide and endothelium-derived hyperpolarizing factor in human arteries and veins. AB - BACKGROUND: We have investigated and compared nitric oxide (NO) release and endothelium-derived hyperpolarizing factor (EDHF)-mediated hyperpolarization in the human internal mammary artery (IMA), radial artery (RA), saphenous vein (SV), and coronary artery. MATERIALS AND METHODS: Vessel segments taken from coronary artery bypass grafting or heart transplantation patients were placed in an organ chamber. NO-sensitive electrode or intracellular glass microelectrode was used to study NO or EDHF in response to acetylcholine (ACh) and bradykinin (BK). RESULTS: The resting membrane potential of the smooth muscle cells of IMA, RA, and SV was 58 +/- 0.84 (n = 61), -61 +/- 1.3 mV (n = 46, p = 0.03), and -62 +/- 0.9 mV (n = 23, p = 0.0001) respectively. BK- (10(-7) M) induced EDHF-mediated hyperpolarization (-10.9 +/- 1.5 mV, n = 7) in the IMA was significantly greater than that in RA (-5.8 +/- 0.9 mV, n = 6, p = 0.04) and SV (-5.1 +/- 0.5 mV, n = 8, p < 0.01). The basal release of NO in IMA (16.8 +/- 1.9 nM) was significantly higher than that in RA (11.1 +/- 1.0 nM, n = 12, p = 0.02) and in SV (9.9 +/- 2.8 nM, n = 13, p < 0.001). The stimulated release of NO to BK in IMA was significantly greater than that in RA (44.3 +/- 4.0 vs 25.8 +/- 3.6 nM, n = 8, p = 0.004). The duration of NO release was longer in IMA than in RA or in SV. CONCLUSIONS: The basal and stimulated release of NO and EDHF-mediated hyperpolarization in the IMA are significantly greater than that in the RA and SV. EDHF exists in all these human vessels. This study reveals the differences among human vessels regarding the endothelial function that have implications in vasospasm, coronary protection, or long-term graft patency. PMID- 12546080 TI - Oxidative stress, nitric oxide, and vascular disease. AB - Superoxide (O2-) is a key risk factor for cardiovascular disease (CVD), including atherogenesis, reperfusion injury, angina, restenosis following balloon angioplasty, and vein graft failure. Axiomatically, O2- reacts with nitric oxide (NO) to form peroxynitrite (ONOO) resulting in a depletion of endogenous vascular NO, which is now firmly associated with CVD. Furthermore, risk factors for CVD, in particular diabetes mellitus (DM), dyslipidemia, and hyperhomocysteinemia are all associated with oxidative stress OS. Antioxidant therapies, including the gene transfer of antioxidant enzymes, are potentially valuable in the treatment of CVD. PMID- 12546081 TI - Adrenomedullin: a cardiac depressant factor in septic shock. AB - Despite intensive research, septic shock is still the most common cause of death in surgical intensive care, and its incidence keeps increasing. No curative treatment is yet available. The critical aspect of septic shock is the refractory hypotension that develops during its late phase which leads to a progressive deterioration of cell and organ functions, and in most instances, death. During septic shock, following the overproduction of cytokines, many factors such as nitric oxide and adrenomedullin (ADM) are produced in abnormally large quantities, but our understanding of their contribution to the pathophysiology of sepsis is limited. Here we show that adrenomedullin (22-52), an adrenomedullin receptor antagonist, improves the contractility of myocytes isolated from lipopolysaccharide (LPS)-treated rats, whereas in normal myocytes, adrenomedullin, acting through an adrenomedullin (22-52) sensitive receptor, decreases their contractility. In addition, adrenomedullin antiserum and inducible nitric oxide (NO) synthase inhibitor improve the survival of LPS treated rats. The data indicate that adrenomedullin is a cardiac depressant factor, which along with NO precipitates ventricular failure during septic shock. PMID- 12546082 TI - Nitric oxide and cyclic GMP as pro- and anti-apoptotic agents. AB - BACKGROUND: Previously we showed that atrial natriuretic peptide (ANP) increases cGMP production in PC12 (sympathetic-neuron-like) cells, cGMP elevations increase survival of hippocampal neurons during glutamate toxicity and ANP-induced cGMP elevations prolongs survival of stressed PC12 cells, all suggesting cGMP mediates anti-apoptotic/pro-survival effects in neural cells. AIM: The objective was to use a new technology, capillary electrophoresis-laser-induced-fluorescence detector (CE-LIF) to accurately measure nitric oxide (NO)-induced stimulation and ANP/cGMP-induced inhibition of apoptotic DNA fragmentation in PC12 and NG108-15 (cholinergic-neuron-like) cells. METHODS: Apoptotic DNA fragmentation was quantified by CE-LIF. RESULTS: Sodium nitroprusside (SNP, 0.1-1.0 mM, 24 hours), NO donor, increased apoptotic DNA fragmentation in NG108-15 cells, but not PC12 cells (both with serum). In serum-deprived PC12 cells, ANP at 1, 10 and 100 nM inhibited apoptotic DNA fragmentation by 75.8%, 84.7%, and 94.1%, respectively. CONCLUSIONS: The data show that NO at higher levels induces apoptosis in NG108-15 cells, but not PC12 cells, indicating differences in susceptibility to NO-induced toxicity, and that ANP-induced cGMP elevation is a potent and effective inhibitor of apoptosis in PC12 cells. The data suggest that NO-induced cGMP elevations in certain neural cells (e.g. PC12 cells) provide a protective (anti-apoptotic) mechanism that counter-balances the pro-apoptotic actions of NO, thus helping to limit damage caused by NO. PMID- 12546083 TI - A mechanosensitive cation channel in endothelial cells. AB - A mechanosensitive Ca2+-permeable channel is present in vascular endothelial cells. The activity of this channel increases in response to hemodynamic blood flow. Recently, it has been found that the activity of this channel may be regulated by cGMP through a protein kinase G-dependent pathway. Inhibition of the channel by cGMP abolishes the Ca2+ influx elicited by flow. Several inhibitors of the cation channel including Gd3+, Ni2+, and SK&F-96365 also inhibit the Ca2+ influx due to flow stimulation. These data suggest that a mechanosensitive cation channel is the primary pathway mediating the flow-induced Ca2+ entry in vascular endothelial cells. PMID- 12546084 TI - Elevation of arterial potassium during acute systemic hypoxia is abolished by alkalosis. AB - BACKGROUND: Small elevations in plasma potassium evoke vasodilation in the peripheral circulation. Systemic hypoxia elevates arterial potassium and also modifies arterial pH. AIMS: We examined the interaction between pH and potassium in blood during systemic hypoxia and the effect of pH on the uptake/release of potassium in the peripheral tissues. METHODS: Anesthetized dogs were ventilated with air plus oxygen for normoxia or air plus nitrogen for hypoxia. Some animals received intravenous sodium bicarbonate to elevate pH by 0.1 units. Arterial plasma potassium concentration was measured in normoxia and hypoxia. A rat gracilis muscle was perfused with normoxic Krebs buffer and the potassium content of the venous outflow was compared during perfusion at pH 7.4, 6.8, or 7.8. RESULTS: In dogs with an arterial pH of 7.40-7.45, systemic hypoxia elevated the arterial potassium by 1 mmol/L. An arterial pH of 7.55 did not alter the basal potassium concentration, but it abolished the hypoxia-induced increase. In rat muscle, reduction of the perfusate pH from 7.4 to 6.8 reduced arterial perfusion pressure from 8.73 to 7.32 kPa and venous potassium from 6.6 to 5.2 mM. Elevation of perfusate pH to 7.8 decreased the arterial perfusion pressure from 8.44 to 6.95 kPa but did not affect venous potassium. CONCLUSIONS: The hypoxia-induced elevation of arterial potassium is abolished by increasing the pH to 7.55. This is not due to enhanced potassium uptake into peripheral tissues at high pH. Red blood cells are suggested as the most likely source of the potassium released in hypoxia. PMID- 12546085 TI - Endothelium-dependent and -independent coronary relaxation induced by urocortin. AB - Urocortin, a newly identified polypeptide, possesses cardiac effects. However, the underlying mechanism of its coronary action is still unclear. In the present study we investigated the possible role of endothelial nitric oxide and Ba2+ sensitive K+ channels in the endothelium-dependent relaxant response to urocortin in the isolated rat left anterior descending coronary arteries. Changes of vessel tone were measured in microvessel myographs. Urocortin produced both endothelium dependent and -independent relaxation with IC50 of 2.52 nM and 16.5 nM, respectively. Denuation of endothelium decreased the relaxing potency of urocortin. In the endothelium-intact rings pretreated with 100 microM N(G)-nitro L-arginine methyl ester (L-NAME) or 10 microM 1H-[1,2,4]oxadiazolo[4,2 alpha]quinoxalin-1-one (ODQ), the urocortin-induced relaxation was similar to that observed in endothelium-denuded rings. The relaxant response to urocortin was markedly reduced in endothelium-intact rings preconstricted by 35 mM K+. Pretreatment with 100 microM BaCl2 significantly reduced urocortin-induced relaxation without an effect on the maximum relaxation. Combined treatment with BaCl2 plus L-NAME did not produce additive inhibition. In contrast, BaCl2 did not alter urocortin-induced relaxation in the endothelium-denuded rings. In the endothelium-denuded rings, BaCl2 at 100 microM also inhibited nitric oxide donor induced relaxation. In conclusion, our results suggest that urocortin-induced endothelium-dependent relaxation of rat coronary arteries is primarily mediated by endothelial nitric oxide and subsequent activation of Ba2+-sensitive K+ channels. The urocortin-induced endothelium-dependent relaxation appears to be cyclic GMP-dependent. PMID- 12546086 TI - Therapeutic angiogenesis for coronary artery disease. AB - Therapeutic angiogenesis may be a realistic approach in treating ischemic heart disease. VEGF is a major angiogenic factor involved in physiological as well as pathological angiogenesis. The ability of VEGF to promote angiogenesis in animal and clinical studies has been studied extensively. However, it is becoming clear that VEGF alone may not be sufficient to effectively complete the angiogenesis process. The use of more than one growth factor may be more pertinent in creating a sustainable angiogenic effect with clinically significant outcome. The challenge is to find complementary partners in angiogenesis to better affect the outcome of the process. To this end, we have been studying the effects of other angiogenic factors such as angiopoietin-1 (Ang-1) in a chronic ischemic porcine model. Single intramyocardial introduction of adenovirus-mediated gene transfer of Ang-1 into the left ventricle free wall has been found to enhance angiogenesis by augmenting the formation of new capillaries that manifested in improved total blood flow in the myocardium. A combined therapeutic angiogenesis study involving VEGF and Ang-1 is currently underway. Due to their unique complementary properties, it is expected that the combination will not merely enhance angiogenesis but will also lead to healthy and mature vascular network in the ischemic myocardium. PMID- 12546087 TI - Nitric oxide and atherosclerotic lesion progression: an overview. AB - There is a growing interest regarding the complex pathophysiological relationship between nitric oxide (NO) and the development of atherosclerosis. The endothelial damage induced by atherogenesis may lead to the reduction in concentration or activity both of inducible and endothelial NO synthase with subsequent impaired release of NO. Moreover, impaired NO diffusion from endothelium to vascular smooth muscle cells is followed by decreased sensitivity to its vasodilator action. Finally, an important mechanism would be a local enhanced degradation of NO by increased generation of reactive oxygen species and other free radicals with subsequent cascade of oxidation-sensitive mechanisms in the arterial wall. Therefore, one target for new drugs should be the restoration of NO-mediated signaling pathways in atherosclerotic arteries. Such novel therapeutic strategies may include administration of L-arginine, the precursor of NO, as well as antioxidants, NO donors, and tissue-specific gene-therapy approaches. PMID- 12546088 TI - Biological mechanisms influencing the function of the aortic root. AB - Optimal function of the aortic root relies upon the ability of its component structures to move in a coordinated fashion. Some of the cells that make up the structures of the aortic root have been shown to contain nerves, receptors, and contractile elements. The ability to contract or relax may contribute to the successful function of the valve by allowing it to move in a coordinated manner in response to biological stimuli. It is known that cusp tissue receives primary, sensory, and autonomic nerves, suggesting a role for neuronal regulation of cusp function. In addition, cusp tissue has been shown to express a wide variety of receptors and to contract to a range of common vasoactive agents. The cells that constitute the valve have also shown secretory and proliferative responses. The biological signals that mediate the cross-talk between the different parts of the root have not been established. This review will examine the mechanisms that have been documented to be present and to assess their potential contribution in affecting aortic valve function. PMID- 12546089 TI - Root coverage of advanced gingival recession: a comparative study between acellular dermal matrix allograft and subepithelial connective tissue grafts. AB - BACKGROUND: Acellular dermal matrix allograft (ADMA) has successfully been applied as a substitute for free connective tissue grafts (CTG) in various periodontal procedures, including root coverage. The purpose of this study was to clinically compare the efficiency of ADMA and CTG in the treatment of gingival recessions > or = 4 mm. METHODS: Seven patients with bilateral recession lesions participated. Fourteen teeth presenting gingival recessions > or = 4 mm were randomly treated with ADMA or CTG covered by coronally advanced flaps. Recession, probing depth, and width of keratinized tissue were measured preoperatively and 12 months postoperatively. Changes in these clinical parameters were calculated within and compared between groups and analyzed statistically. RESULTS: Baseline recession, probing depth, and keratinized tissue width were similar for both groups. At 12 months, root coverage gain was 4.57 mm (89.1%) versus 4.29 mm (88.7%) (P = NS), and keratinized tissue gain was 0.86 mm (36%) versus 2.14 mm (107%) (P < 0.05) for ADMA and CTG, respectively. Probing depth remained unchanged (0.22 mm/0 mm), with no difference between the groups. CONCLUSIONS: Recession defects may be covered using ADMA or CTG, with no practical difference. However, CTG results in significantly greater gain of keratinized gingiva. PMID- 12546090 TI - A 3-year longitudinal evaluation of subpedicle free connective tissue graft for gingival recession coverage. AB - BACKGROUND: The purpose of this study was to longitudinally evaluate, over a 3 year period, the reduction of gingival recession through use of a subepithelial free connective tissue graft placed under a coronally advanced partial-thickness pedicle flap. METHODS: Twenty-one buccal recession defects (mean 3.67 mm; range 3 to 4.5 mm; Miller Class I, II, and III) in 15 patients were treated using this technique. Amount of gingival recession (GR), clinical attachment loss (CAL), and width of keratinized gingiva (WKG) were followed for 3 years after surgery. The measurements were performed at presurgery, and 1, 3, 6, 12, 18, 24, and 36 months postsurgery. RESULTS: GR decreased from 3.67 +/- 0.58 mm at baseline to 0.33 +/- 0.43 mm at 36 months, representing a reduction of 3.33 mm, corresponding to 91.28% mean root coverage. CAL was significantly decreased at 36 months from 5.26 +/- 0.77 mm to 2.14 +/- 0.57 mm. At 36 months, 3.12 mm of attachment gain was obtained. WKG significantly increased after 36 months (1.95 mm). GR, CAL, and WKG had the most positive outcomes at 12 months and were maintained at stable levels throughout the 36-month observation period. CONCLUSIONS: These results indicate that the connective tissue graft with a partial thickness coronal advancement pedicle is a predictable method for root coverage and, provided that optimal maintenance care is provided, the clinical outcomes gained by this technique can be well maintained. PMID- 12546091 TI - Coverage of previously carious roots is as predictable a procedure as coverage of intact roots. AB - BACKGROUND: Coverage of roots exposed by gingival recession is one of the main objectives of periodontal reconstructive surgery. A variety of highly predictable and esthetically acceptable mucogingival grafting procedures are available for treating intact root surfaces. One of the indications for root coverage procedures is prevention of root caries. However, little is known about the ability to treat previously carious roots by mucogingival grafting, and even less is known about the outcome of such treatment. METHODS: Sixty patients with gingival recession were treated with subepithelial connective tissue grafts. During initial examination, 33 intact teeth with longstanding gingival recessions of 4.09 +/- 0.16 mm (mean +/- SEM) and 27 teeth with carious roots (restored and unrestored) and longstanding gingival recessions of 3.44 +/- 0.22 mm (mean +/- SEM) were found. Prior to grafting, any carious dentin and plastic restorations were removed. The exposed roots were thoroughly planed and covered by a subepithelial connective tissue graft without any further root treatment or conditioning. Patients were evaluated periodically between 1 and 6 years. Residual recession, defect coverage, and caries incidence were assessed. RESULTS: Recession was reduced to a similar extent on all teeth whether root caries was present (0.31 +/- 0.11 mm) or not (0.15 +/- 0.04 mm). This resulted in comparable defect coverages of 92.41 +/- 2.38% for previously carious roots and 97.46 +/- 0.79% for intact roots. In 44 cases, complete coverage of the recession was achieved. The results were stable, and no further recessions or recurrent caries were found during the periodic evaluations. CONCLUSIONS: These results indicate that coverage of previously carious roots with subepithelial connective tissue grafts is a very predictable procedure with results similar to those found on intact roots. This procedure may provide a definitive biological alternative for conventional restorative treatment modalities for root caries. Moreover, the results are more esthetic, biologically acceptable, and maintainable. PMID- 12546092 TI - Predicting tooth loss during a population-based study: role of attachment level in the presence of other dental conditions. AB - BACKGROUND: Our objectives were to quantify: 1) the association between incident tooth loss and prior periodontal attachment level; and 2) the contribution to tooth loss made by non-periodontal conditions in increasingly periodontally involved teeth. METHODS: The Florida Dental Care Study was a prospective cohort study of persons who at baseline had at least 1 tooth and were 45 years or older. In-person interviews and clinical examinations were conducted at baseline, and at 24 and 48 months, with telephone interviews at 6-month intervals in between. A regression model was used to simultaneously quantify tooth-specific predictors of tooth loss, with person-level factors taken into account. RESULTS: Of the 687 persons who participated for a 48-month clinical examination, 36% lost 1 or more teeth during follow-up, and 5.0% of all teeth were lost. Attachment level up to 2 years before tooth loss was strongly predictive of incident tooth loss, with increases in risk for each millimeter in attachment loss. Certain other tooth specific conditions (tooth mobility, bulk restoration fracture, decayed surfaces, filled surfaces, tooth type and arch location, root fragment) were strongly and independently associated with increased risk for tooth loss, while others were not (prosthetic crown coverage, cusp fracture, root surface defect). Propensity to choose extraction over other treatment alternatives, as reported by participants at baseline, was also strongly predictive of tooth loss. CONCLUSIONS: Increasingly severe attachment level was consistently associated with an increased risk for tooth loss in this sociodemographically diverse sample, with or without other tooth-specific conditions taken into account. PMID- 12546093 TI - Detection of human viruses in patients with chronic periodontitis and the relationship between viruses and clinical parameters. AB - BACKGROUND: Recent studies have demonstrated that various human viruses, especially cytomegalovirus (HCMV), Epstein-Barr virus type-1 (EBV-1), and herpes simplex virus (HSV), seem to play a part in the pathogenesis of human periodontitis. Little information is available on the relationship between these viruses and clinical periodontal parameters in patients with chronic periodontitis. This study examined the occurrence of HCMV, EBV-1, and HSV in patients with chronic periodontitis and the relationship between these viruses and clinical parameters. METHODS: A nested polymerase chain reaction (PCR) method determined the presence of HCMV, EBV-1, and HSV. Subgingival plaque samples from 30 patients with chronic periodontitis and 21 randomly selected healthy controls were collected by paper points, and clinical measurements were recorded from both sampling sites and entire dentition. The following indices were measured: plaque index (PI), gingival index (GI), probing depth (PD), and clinical attachment loss (CAL). RESULTS: HCMV was detected in 44.3% of chronic periodontitis patients and 14.3% of healthy persons (P < 0.05); EBV-1 in 16.7% of chronic periodontitis patients and 14.3% of healthy persons (P = 1.00); and HSV in 6.7% of chronic periodontitis patients and in no healthy persons. HCMV and EBV-1 detected and undetected sites in patients with periodontitis showed statistically significant differences in sampling clinical depth (SPD) and sampling clinical attachment loss (SCAL). Differences in the measurements of PI of entire dentition and GI of entire dentition between HSV detected and undetected sites were statistically significant. CONCLUSIONS: Findings of the present study confirm the frequent presence of HCMV in crevicular samples of chronic periodontitis lesions, and suggest a strong relationship between the presence of HCMV and EBV-1 in subgingival areas and the measurements of probing depth and probing attachment loss. PMID- 12546094 TI - IgG subclass specific antibody response to periodontopathic organisms in HIV positive patients. AB - BACKGROUND: We previously reported an increased rate of progression of periodontal disease over an 18-month period in human immunodeficiency virus (HIV) positive subjects compared to controls. The mechanism for disease progression and rapid tissue loss was unknown. Data on the microbiological studies failed to show any significant difference in the microbial characteristics of the periodontal lesions in HIV-positive patients compared to HIV-negative controls. Immunological analysis had identified neutrophils as an important component of the host defense against periodontal infection, especially against rapid tissue loss. Serum IgG reactivities to periodontal pathogens in HIV-positive patients with periodontitis were reduced. Other data provided circumstantial evidence to suggest that IgG subclass (IgG2) specific antibody might assist bacterial clearing in periodontal infection. The aim of the current study was to examine the specific IgG subclass antibody response to a panel of periodontopathic organisms: Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella Intermedia (Pi), Fusobacterium nucleatum (Fn), Campylobacter rectus (Cr), and Bacteroides forsythus (Bf) in HIV-positive patients compared to HIV-negative controls. METHODS: Sera from 120 HIV-positive patients (40 periodontitis, 69 gingivitis, and 11 no oral diseases) were tested for IgG subclass specific antibody response to the above listed 6 organisms using enzyme-linked immunosorbent assay. Data were compared with those obtained from 40 HIV-negative control subjects (35 periodontitis, 2 gingivitis, and 3 no oral diseases). RESULTS: In the HIV positive group, a consistently high response rate was found in IgG1 to all the bacteria tested. In addition, high levels of IgG3 and IgG4 to Pg and IgG1 and IgG2 to Pi were also present. However, no significant difference was detected among the periodontitis, gingivitis, and no oral disease subgroups. When the periodontitis patients from the HIV-positive group were compared to the HIV negative group, no difference in the antibody levels and response rates was noted. CONCLUSION: We conclude that in HIV-positive patients, the specific IgG subclass antibody response to periodontopathic organisms was similar to that of HIV-negative subjects. PMID- 12546095 TI - Osseous reconstruction using a membrane barrier following marginal mandibulectomy: an animal pilot study. AB - BACKGROUND: Invasive and malignant tumors of the oral soft tissues adjacent to the mandible are often treated with partial resection of the osseous structure adjacent to the lesion (marginal mandibulectomy). Autogenous bone graft and composite bone grafts are being used to fill the osseous defects with various degrees of success. The aim of the present study was to explore the utilization of membrane barriers and the principle of guided bone regeneration to negotiate these defects. METHODS: Following the removal of P1, P2, and P3, experimental bilateral marginal mandibulectomy defects were created in 4 adult dogs. The bone segments (measuring 25 mm x 15 mm) were then removed. Each side was then randomly selected for either experimental (titanium-reinforced expanded polytetrafluoroethylene membrane, [ePTFE-TR]) or control (repositioning flaps) treatment. Postoperatively, the animals were put on soft diet, antibiotics, and analgesics. Sutures were removed under light sedation after 4 weeks, and the area was left to heal and mature for 4 to 6 months (mean 5.3 months). The animals were then sacrificed, and block sections of the mandible were obtained for macroscopic and histological evaluation. RESULTS: The size of the residual defect (the vertical distance between the most apical depression in the ridge and the horizontal line connecting the free gingival margins of the proximal teeth) in the experimental sites (6.10 +/- 1.00 mm) was much smaller compared to the controls (10.65 +/- 0.82 mm), which was statistically significant (P = 0.0127). Histomorphometric measurements of new bone formation (NBF) revealed a similar pattern: for the experimental sites, NBF was 8.08 +/- 0.85 mm compared to 4.99 +/ 0.61 mm in the controls. These differences were also statistically significant (P = 0.0257). CONCLUSIONS: A regenerative approach to large mandibular osseous defects has been described. If this new treatment modality is further substantiated in other independent studies, it might prove a useful tool in restoring the lost osseous structure associated with marginal mandibulectomy procedures. PMID- 12546096 TI - The effect of safflower seed extract on periodontal healing of 1-wall intrabony defects in beagle dogs. AB - BACKGROUND: Recent interest in naturally based products has increased. Various herbal extracts are known to have a variety of medicinal properties. Among the various natural medicines, safflower seeds have beneficial effects on various bone diseases such as bone fracture, osteoporosis, and osteodysplasia. In addition, they are known to have anti-inflammatory effects. The objective of this study was to evaluate the effect of a safflower seed extract (SSE) on the regeneration of periodontal tissue in a preclinical 1-wall model in dogs. METHODS: Preclinical 1-wall periodontal defects were surgically created in the mesial aspect of the maxillary third and mandibular fourth premolar and in the distal aspect of the maxillary first and mandibular second premolar, and were randomly assigned to receive SSE/collagen (SSE/Col), phosphate-buffered saline/collagen (buffer control), or root planing only (surgical control). The created 1-wall defect configuration was 4 mm in depth by 4 mm in width. We selected the segment showing the best activity to the osteoblast cells that was sensitive to the formation of calcified nodules among the SSE fractions extracted from various organic solvents. The animals were euthanized at 8 weeks postsurgery, and block sections of the defects were collected for histologic and histometric analysis. RESULTS: The junctional epithelium migration did not show any statistically significant differences among the treatments. In connective tissue adhesion, the SSE/Col group and the buffer control group showed significant differences compared to the surgical control group. New cementum averaged 3.84 +/- 0.57 mm, 3.75 +/- 0.24 mm, and 1.53 +/- 1.22 mm for the SSE/Col group, the buffer control group, and the surgical control group, respectively, with the SSE/Col and buffer control groups significantly different from the surgical control group (P < 0.05). The amount of intrabony cementum in the SSE/Col group was significantly different (P < 0.01) from the surgical control group, but the amount of suprabony cementum did not demonstrate any statistical difference between the different treatments. The amount of new alveolar bone averaged 2.93 +/- 0.70 mm, 2.10 +/- 0.63 mm, and 1.20 +/- 0.65 mm for the SSE/Col group, the buffer control group, and the surgical control group, respectively. The difference in alveolar bone regeneration between the SSE/Col group and the surgical control group was significantly different (P < 0.01). Root resorption was often observed, but no ankylosis was present. CONCLUSION: Wound conditioning with safflower seed extracts may contribute to bone formation but appears to have unpredictable potential for stimulating periodontal regeneration including new cementum. PMID- 12546097 TI - Effects of basic fibroblast growth factor on human gingival epithelial cells. AB - BACKGROUND: Our previous reports found that basic fibroblast growth factor (FGF 2; bFGF) influences the proliferation and extracellular matrix production of periodontal ligament (PDL) cells. In this study, we examined FGF-2 expression in gingival epithelium and the effect of FGF-2 on proliferative responses by gingival epithelial (GE) cells. METHODS: Human GE cells were isolated from healthy gingival epithelium, and the mRNA expression of FGF-2 and FGF receptors (FGFRs) was examined by reverse transcription-polymerase chain reaction (RT-PCR). The distribution of FGF-2 in gingival tissues was detected by immunohistological analysis using the monoclonal antibody for human recombinant FGF-2, which was newly established and designated as BF-2. Further, the proliferative responses of GE cells to FGF-2 were investigated by measuring [3H]-thymidine uptake. RESULTS: RT-PCR revealed that GE cells express FGFR-1, FGFR-2, FGFR-3, and FGFR-4 mRNA; however, not that of FGF-2. Employing immunohistochemical staining with BF-2, FGF 2 was observed localized in the intercellular spaces of gingival epithelium, though not in the cytoplasm of epithelial cells. Interestingly, staining by BF-2 in the intercellular spaces was diminished after treatment of the tissue sections with heparitinase. Further, an in vitro analysis revealed that FGF-2 enhanced the proliferative responses of human GE cells. However, costimulation with fetal calf serum inhibited the FGF-2-induced proliferation of GE cells, whereas the same costimulation synergistically enhanced FGF-2-induced PDL cell proliferation. CONCLUSIONS: FGF-2 is anchored in the intercellular spaces of gingival epithelium via heparansulfate and may regulate the growth and cytodifferentiation of GE cells via cell-type specific receptors. PMID- 12546098 TI - The cost-effectiveness of interleukin-1 genetic testing for periodontal disease. AB - BACKGROUND: A genetic test for a composite interleukin-1 (IL-1) genotype is being marketed to predict risk for progression of periodontal disease. The objective of this study was to determine the clinical scenario required to produce cost effective results with the use of IL-1 testing to identify high-risk patients. METHODS: A disease simulation model was developed using decision-analytic techniques and a 30-year time frame. RESULTS: Using different modeling scenarios, the genetic test produced results ranging from cost savings of $830,140 and 52.8 fewer cases of severe periodontitis to increased costs of $300,430 and 3.6 additional cases of severe periodontitis (per 1,000 patients). Three parameters in the analysis were highly influential: 1) the compliance rate for maintenance therapy in test positive versus non-tested patients; 2) the effectiveness of non surgical therapy; and 3) the relative risk of disease progression for test positive patients. CONCLUSION: The model produced a wide range of outcomes reflecting our incomplete understanding of the biology, optimal treatment, and genetic susceptibility of periodontal diseases. However, the model demonstrates that three clinical parameters are highly influential in determining if IL-1 testing can be implemented in a primary care setting in a cost-effective manner. PMID- 12546099 TI - The prognostic value of several periodontal factors measured as radiographic bone level variation: a 10-year retrospective multilevel analysis of treated and maintained periodontal patients. AB - BACKGROUND: Assigning a prognosis to a periodontal patient is one of the greatest challenges in clinical practice. Many different factors can affect the result of periodontal therapy. The purpose of this study was to evaluate the prognostic value of some clinical, genetic, and radiographic variables in predicting bone level variation in periodontal patients (aged 40 to 60) treated and maintained for 10 years. METHODS: Sixty consecutive non-smoking patients (mean age 46.77 +/- 4.96) with moderate to severe chronic periodontitis were treated with scaling and root planing (SRP). Some patients also underwent additional surgical treatments. All patients were maintained in the same private practice for 10 years. At baseline (T0) and at least 10 years later (T2), the following clinical variables were evaluated: probing depth (PD), tooth mobility (TM), presence of prosthetic restorations (PR), and molar teeth (MT). In addition, radiographic measurements were taken of the mesial and distal distances from the cemento-enamel junction (CEJ) to the bottom of the defect (BD), to the bone crest (BC), and to the root apex (RA). At T2, a genetic test to determine the IL-1 genotype and genetic susceptibility for severe periodontal disease was performed for all 60 patients. Based on the results of this assay, the patients were categorized as IL-1 genotype positive (G+) or negative (G-). The differences between the bone levels measured at T0 and T2 (ABD), indicating the bone level variation, was used as the outcome variable. Different predictor variables were then tested using a 3-level statistical model (multilevel statistical analysis; patient, tooth, and site level). At the patient level these were: age, gender, and interaction between mean bone loss and the IL-1 genotype (mean CEJ-BD(T0) x IL-1 genotype). At the tooth level the variables were: TM(T0), PR(T0), MT(T0); and at the site level the evaluated factors were: the infrabony component of the defect (CEJ-BD(T0) - CEJ BC(T0), PD(T0), bone level (CEJ-BD(T0)), and the residual supporting bone (BD RA(T0)). RESULTS: Among the considered predictor parameters, the following were significantly associated with the outcome variable: 1) mean CEJ-BD(T0) x IL-1 genotype (P = 0.0019); 2) TM(T0) (P < 0.0000); 3) CEJ-BD(T0) (P < 0.0000); 4) CEJ BD(T0) - CEJ-BC(T0) (P < 0.0000); 5) PD(T0) (P = 0.0010). Deeper probing depths at a site and tooth mobility at baseline were associated with worst prognosis. Greater CEJ-BD(T0) distance and infrabony component at a site at baseline were associated with a better prognosis. The interaction between mean CEJ-BD measurement at baseline and IL-1 genotype was significantly associated both with a good or a poor prognosis. The other parameters evaluated - age, gender, presence of molars and prosthetic restorations, and residual supporting bone - were not significantly associated with bone level variation. CONCLUSIONS: Within the scope of this study design, many traditional prognostic factors were ineffective in predicting future bone level variation and therefore were of no prognostic value. Conversely, a few specific factors at each level emerged as valuable prognostic factors. At the patient level, the prognostic factor was initial mean bone level in conjunction with a positive IL-1 genotype. At the tooth level, the prognostic factor was tooth mobility. At the site level, the significant prognostic factors were initial bone level at a site, the infrabony component of a defect, and initial probing depth at a site. The use of these factors may be of value to clinicians as predictors of bone level variation when assigning a prognosis to a patient, a tooth, or a site. PMID- 12546100 TI - Effect of surgical implantation of recombinant human bone morphogenetic protein-2 in a bioabsorbable collagen sponge or calcium phosphate putty carrier in intrabony periodontal defects in the baboon. AB - BACKGROUND: Recombinant human bone morphogenetic protein-2 (rhBMP-2) in a proper carrier has been shown to induce clinically relevant bone formation for several oral/maxillofacial and periodontal indications and to stimulate regeneration of the periodontal attachment. The objective of this study is to evaluate regeneration of alveolar bone, cementum, periodontal ligament, and associated root resorption and ankylosis following surgical implantation of rhBMP-2 in an absorbable collagen sponge (ACS) or a calcium phosphate putty (alphaBSM) carrier in 3-wall intrabony periodontal defects in the baboon. METHODS: rhBMP-2/ACS and rhBMP-2/alphaBSM were implanted in surgically produced, maxillary and mandibular, large size, 3-wall intrabony defects in 4 baboons. Contralateral jaw quadrants were implanted with buffer/ACS, buffer/ alphaBSM, or served as sham-operated surgical controls. Treatments were allocated to left and right, maxillary and mandibular, jaw quadrants following a randomization schedule. Four months following implantation, block biopsies of defect sites were obtained, processed, and subjected to histologic and histometric analysis. RESULTS: Defect sites receiving rhBMP-2/ACS and rhBMP-2/alphaBSM demonstrated significantly greater regeneration than controls. No significant differences were observed between defect sites receiving rhBMP-2/ACS or rhBMP-2/alphaBSM regarding epithelial migration and connective tissue attachment and new bone formation. However, rhBMP 2/ACS supported significantly greater new cementum formation. Ankylosis or root resorption were not observed. CONCLUSIONS: The results of this study support the use of rhBMP-2 to enhance periodontal regeneration of intrabony periodontal defects. While this novel technology holds promise, refinement in carrier systems may provide the key to enhancement of the regenerative potential. PMID- 12546101 TI - Isotypic antibody response to plaque anaerobes in periodontal disease. AB - BACKGROUND: It has been suggested that locally produced immunoglobulin (Ig)A could be more protective than IgG and that there could be a relationship between crevicular fluid-specific IgA levels and the onset of periodontal disease. This study was designed to investigate this hypothesis regarding specific immune responses towards 4 plaque anaerobes in gingival crevicular fluid and saliva from patients with periodontopathies and controls. METHODS: Gingival crevicular fluid (GCF) and whole saliva were collected from 35 adults with periodontitis and 24 periodontally healthy adults (controls). Antigens were extracted from Actinomyces actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, and Fusobacterium nucleatum and used to set up specific enzyme-linked immunosorbent assay (ELISA) tests to assess IgA and IgG levels to these microorganisms in the fluids collected. RESULTS: The crevicular fluid of periodontitis patients contained significantly higher levels of IgG to the 4 microorganisms tested than that of controls (P < 10(-6) for all comparisons). IgA levels to the 4 bacteria were statistically significantly much higher in control crevicular fluid (P < 10( 7) for all comparisons). Controls also had statistically significantly higher levels of specific salivary IgA than patients (P < 0.02 for all comparisons). CONCLUSIONS: These data support the potentially protective role of specific IgA directed to oral microorganisms involved in the onset and development of periodontal disease. PMID- 12546102 TI - Pyostomatitis vegetans as an early sign of reactivation of Crohn's disease: a case report. AB - BACKGROUND: The structures and lining of the oral cavity are an accessible mirror reflecting the health of an individual. They may play an important role in detecting and monitoring systemic diseases. Certain conditions that primarily affect other regions of the body may cause signs and symptoms in the mouth, either as the initial presenting symptom or in conjunction with other systemic manifestations. METHODS: Pyostomatitis vegetans, which occurred in a patient with a history of Crohn's disease, was diagnosed as an early sign of disease reactivation. RESULTS: The patient was treated with a topical fluorinated corticosteroid gel, which led to the improvement of the oral lesions. However, the oral lesions continued to recur over a period of several months until the underlying inflammatory bowel disease came under control. CONCLUSIONS: This report emphasizes the relationship of pyostomatitis vegetans to Inflammatory bowel disease and the importance of the oral lesions as initial presenting signs of systemic disease or activity. PMID- 12546103 TI - Combined therapy of mineral trioxide aggregate and guided tissue regeneration in the treatment of external root resorption and an associated osseous defect. AB - BACKGROUND: The treatment of external root resorption and associated periodontal defects can be challenging to the most astute clinician. In this case report, a multidisciplinary approach was performed to treat a maxillary central incisor that presented with a sinus tract. An amalgam restoration had been placed approximately 10 years earlier to repair an area of external root resorption. METHODS: A full-thickness mucoperiosteal flap was reflected from teeth #8 to #9. Following degranulation of the area, an amalgam restoration was found on the distal root surface of tooth #8. A 2-wall osseous lesion was also associated with the distal surface of #8. The amalgam was removed and the defect was restored with mineral trioxide aggregate (MTA). The root surface was chemically conditioned with tetracycline, and the osseous defect was grafted with decalcified freeze-dried bone allograft (DFDBA) and a calcium sulfate barrier. RESULTS: An 8 mm gain in clinical attachment, as well as an increase in radiodensity, was noted on the distal surface of tooth #8 at 15 months postsurgery. The patient was also asymptomatic, with no clinical signs of inflammation present. CONCLUSIONS: A combined approach utilizing MTA for root surface repair, and DFDBA and calcium sulfate to address an associated osseous lesion, appears to be a viable modality in the treatment of chronic endodontic/periodontal lesions. PMID- 12546104 TI - Alveolar bone upper growth in furcation area using a combined orthodontic regenerative therapy: a case report. AB - BACKGROUND: This case report demonstrates orthodontic and regenerative combined therapy in a 49-year-old male whose right maxillary premolar furcation had a bony defect with poor biologic width resulting from extensive subgingival caries. In these advanced interdisciplinary cases, crown lengthening with periodontal surgery alone does not solve the complex clinical problems. We believe that a combined orthodontic and periodontal regenerative combination therapy offers the best option for achieving a predictable outcome. METHODS: First, regenerative therapy by open debridement with a bioabsorbable synthetic bone graft, bioabsorbable membrane, and minocycline root conditioning was carried out. Eight weeks after initial surgery, orthodontic extrusion was initiated. RESULTS: Radiographs and reentry documentation suggest that the furcation defect associated with poor biologic width was successfully treated. CONCLUSION: This case report demonstrates that orthodontic-regenerative combined therapy can resolve complex clinical problems and enhance predictability. PMID- 12546105 TI - Langerhans' cell histiocytosis diagnosed through periodontal lesions: a case report. AB - BACKGROUND: Langerhans' cell histiocytosis (LCH) is a rare disorder in which a neoplastic proliferation of cells similar to the Langerhans' cell of the epidermis is observed. The disease may affect different organs as solitary or multiple lesions with a complete dissemination in different organs such as the bone (jaws), lung, hypothalamus, skin and mucous membranes, lymph nodes, liver, and other tissues. Although LCH is considered a childhood or juvenile disease, the diagnosis is often made in adults, and in many cases, a juvenile form progresses into adult life. Hand-Schuller-Christian disease (HSCD) is an LCH form in which the typical lesions Involve the cranial bones, the eyes, and the pituitary gland. METHODS: This article describes a case of a patient diagnosed, through periodontal lesions and diabetes insipidus, as having Langerhans' cell histiocytosis. CONCLUSION: As far as LCH lesions appearing in periodontal tissues, the periodontist should be involved in the detection of oral lesions. PMID- 12546106 TI - Histological evaluation of 4 cases of root coverage following a connective tissue graft combined with an enamel matrix derivative preparation. AB - BACKGROUND: The purpose of this report was to evaluate the histological characteristics of 4 clinically successful root coverage procedures performed with a combination of a connective tissue (CT) graft and an enamel matrix derivative (EMD) preparation. METHODS: Four maxillary teeth (3 canines and 1) first premolar) that presented with mucogingival problems and Miller's Class II and III gingival recession were evaluated in the study. Surgical therapy for root coverage consisted of autogenous CT grafts combined with the topical application of EMD. Following healing, treated teeth were extracted with a portion of their buccal attachment apparatus. Two teeth were extracted at 6 months and 2 teeth at 1 year postoperatively. Specimens were prepared for histology and analyzed by light microscopy. RESULTS: All treated teeth presented with clinical signs of successful root coverage. Histological observations of the 4 specimens revealed similar findings. The junctional epithelium was < or = 2 mm long. Apical to the junctional epithelium, dense CT fibers were found in close proximity to the root surface, but in general, no insertion of these fibers into the root was observed. In one 6-month specimen, formation of new cementum and new bone was observed in the most apical end of the grafted area. CONCLUSIONS: The results of this study suggest that a combination of CT grafts and EMD results mainly in an adhesion between the CT and root surface. Some periodontal regeneration may occur in some regions. The development of a long junctional epithelium was not observed with this combined therapy for the treatment of gingival recession. PMID- 12546107 TI - Re: A critical assessment of interleukin-1 (IL-1) genotyping when used in a genetic susceptibility test for severe chronic periodontitis. Greenstein G, Hart TC (2002;73:231-247). PMID- 12546108 TI - What to do until HAART arrives? PMID- 12546109 TI - A new approach to treating patients with MDR-TB? PMID- 12546110 TI - A critical review of diagnostic approaches used in the diagnosis of childhood tuberculosis. AB - SETTING: The diagnosis of tuberculosis (TB) in children is seldom confirmed, and is based mainly on clinical signs, symptoms and special investigations. Various attempts in the form of diagnostic approaches have been made to rationalise this diagnostic process. AIMS: To review and describe published diagnostic approaches aimed at diagnosing mainly intrathoracic tuberculosis in children in developing countries; to compare diagnostic approaches with each other and with bacteriologically confirmed TB; and to describe modifications to the diagnosis of TB in HIV-infected or malnourished children. METHODS: Literature review classified into 1) diagnostic approaches, 2) characteristics used in diagnostic approaches, and 3) studies done to validate diagnostic approaches. RESULTS: Sixteen systems were analysed. Comparison of systems is difficult because characteristic definitions and the ranking of characteristics are not standardised, few studies have been performed to validate these diagnostic approaches, and the gold standard of diagnosis is not practicable in most settings. The minority of systems are adapted for HIV-infected and malnourished patients. RECOMMENDATIONS: Characteristic definitions and ranking of characteristics should be standardised. Any new diagnostic approaches developed should be relevant to developing countries with limited resources, a high burden of tuberculosis, malnutrition and HIV/AIDS and a young population. Studies done to validate diagnostic approaches should be conducted scientifically. PMID- 12546111 TI - Cotrimoxazole prophylaxis in HIV-infected individuals after completing anti tuberculosis treatment in Thyolo, Malawi. AB - SETTING: Thyolo, rural southern Malawi. OBJECTIVES: To determine 1) the proportion who continue with cotrimoxazole prophylaxis for the prevention of opportunistic infections, and 2) the reasons for continuing or stopping prophylaxis, in human immunodeficiency virus (HIV) infected individuals with tuberculosis (TB) who complete anti-tuberculosis treatment. DESIGN: A cross sectional study. METHODS: A questionnaire study of all HIV-infected TB patients who had been registered over a 3-month period to receive anti-tuberculosis treatment and cotrimoxazole prophylaxis and who had completed antituberculosis treatment 3-6 months earlier. RESULTS: Of 82 HIV-infected individuals who were alive at the time of interview, 76 (93%) were continuing with cotrimoxazole and wished to do so indefinitely. The most common reason for continuing the drug was to prevent illness associated with HIV, while the most common reason for stopping was long distances to the health facility. Ninety-six percent of patients received cotrimoxazole free of charge from a health centre. Of those who wished to continue indefinitely, the majority (63%) could not afford to pay for the drug. CONCLUSIONS: In a rural setting, the great majority of HIV-infected individuals continued with cotrimoxazole after completing anti-tuberculosis treatment. Making the drug available and providing it free of charge is essential if it is to remain accessible for longer term prevention. PMID- 12546112 TI - Influence of highly active anti-retroviral therapy (HAART) on the natural history of extra-pulmonary tuberculosis in HIV patients. AB - OBJECTIVE: To determine factors related to survival in acquired immune-deficiency syndrome (AIDS) patients with extra-pulmonary tuberculosis (EPTB), when this condition is the first AIDS-defining disease. DESIGN: A retrospective cohort study of 549 AIDS patients with EPTB as the first AIDS-defining disease. Potential candidates to predict survival were sex, human immunodeficiency virus (HIV) exposure, the coexistence of pulmonary and EPTB at diagnosis, tuberculin skin test, directly observed therapy for tuberculosis (DOT), and highly active anti-retroviral therapy (HAART). The Kaplan-Meier method and Cox regression models were used to assess factors associated with survival. RESULTS: Estimated 3 year survival was 47.0% for those diagnosed before 1993, 72.6% for patients with first AIDS diagnosis during 1995-1996 and 84.6% for those diagnosed after 1996. A negative tuberculin test (hazard ratio [HR] = 1.6, 95% CI 1.1-2.3), not being on DOT (HR 2.2; 95% CI 1.3-3.7) and having pulmonary tuberculosis involvement also (HR 1.3; 95% CI 1.1-1.7) were independently associated with poorer survival. The survival of patients significantly improved after the introduction of HAART (HR 0.4; 95% CI 0.2-0.6). CONCLUSION: The survival of HIV patients with EPTB as their first AIDS-defining disease has substantially improved during the last decade. A negative tuberculin skin test and not receiving DOT are associated with poorer survival among HIV-infected patients whose first AIDS-defining disease is EPTB. PMID- 12546113 TI - Treatment results of DOTS in 1797 Sudanese tuberculosis patients with or without HIV co-infection. AB - SETTING: Consecutive new tuberculosis (TB) patients, from eight states in Sudan, who had never been previously treated for as much as 1 month between 1998 and 2000. OBJECTIVE: To determine the impact of human immunodeficiency virus (HIV) co infection on tuberculosis treatment outcome. DESIGN: All patients presenting with symptoms suggestive of tuberculosis underwent sputum microscopy for acid-fast bacilli (AFB). Treatment is free of charge, and directly observed for all smear positive patients. Treatment outcomes were those defined by the World Health Organization. All patients were tested anonymously for human immunodeficiency virus (HIV) using the Bionor test. RESULTS: Of 10 494 patients suspected of TB and referred for sputum microscopy, 1797 were TB cases; 983 had smear-positive pulmonary tuberculosis, 521 smear-negative pulmonary tuberculosis, and 293 extra pulmonary tuberculosis. Smear-positive cases showed a cure rate of 77.2% and a failure rate of 1%. Smear-negative and extra-pulmonary patients had a completion rate of 79.4%. Cure rates for the smear-positive cases were 68.3% for HIV positive and 77.6% for HIV-negative patients (P = 0.164). Case fatality was significantly higher among HIV-positive (12%) than among HIV-negative cases (1.8%) (OR 7.7, 95% CI 3.51-16.8). CONCLUSION: To date, a relatively low proportion of tuberculosis patients in Sudan also have HIV infection. These patients are substantially more likely to die while on treatment for their tuberculosis, a fact that underlines their need for more comprehensive care if their lives are to be prolonged. In addition, every effort is required to diminish the transmission of HIV infection to prevent the tragedy this infection represents to the community. PMID- 12546114 TI - Clinical indicators of mycobacteraemia in adults admitted to hospital in Blantyre, Malawi. AB - SETTING: Adult medical wards of a central hospital in Blantyre, Malawi. OBJECTIVE: To measure the prevalence and outcome of mycobacteraemia in febrile hospitalised adults, and to determine what proportion could be identified using routine methods; to assess clinical indicators of mycobacteraemia, and the usefulness of a diagnostic trial of anti-tuberculosis treatment. DESIGN: We prospectively examined adults admitted with fever or a history of fever. All had blood cultured for bacteria and mycobacteria, chest X-ray and sputum smears. FINDINGS: Mycobacterium tuberculosis was the commonest cause of blood stream infection (BSI), affecting 57 of 344 patients (17%). In 44 (77%) patients with mycobacteraemia, TB was identified using routine investigations; it was not suspected in six (11%). Strong clinical indicators of mycobacteraemia were anaemia, HIV seropositivity, cough, chronic fever and a clinical diagnosis of AIDS on the day of admission. Of nine patients selected for a therapeutic trial of tuberculosis (TB) treatment, six had mycobacteraemia, of whom five died during the trial. Mortality on short-course chemotherapy, on the TB ward after 1 month was similar whether patients had mycobacteremia (21%) or not (32%). CONCLUSION: TB can be identified with routine methods in most patients with mycobacteraemia. If treated, mycobacteraemia has as good an early outcome as TB without mycobacteraemia. Strengthening of basic facilities is likely to improve detection and treatment of mycobacterial disease. PMID- 12546115 TI - Patients' beliefs: do they affect tuberculosis control? A study in a rural district of South Africa. AB - SETTING: A rural district of South Africa. OBJECTIVES: To describe the beliefs and experiences about tuberculosis of patients and community members and to suggest how these affect presentation to health services and treatment adherence. DESIGN: A descriptive study using structured interviews with individual patients, and focus group interviews with patients and community groups. RESULTS: There is a strong belief in this community that tuberculosis is the result of breaking cultural rules that demand abstinence from sex after the death of a family member and after a woman has a spontaneous abortion. People believe that the resulting disease can only be treated by traditional healers. This delays presentation to hospitals or clinics. There is also a belief in a 'western' type TB that can spread from sufferers or is due to environmental pollution or to smoking or alcohol excesses. A number of factors were cited that influenced adherence to treatment. These included the stigma of TB, the belief that there should be abstinence from sex while on treatment, difficulties accessing health services, long waits and unacceptable health worker attitudes. CONCLUSIONS: With the increasing rates of tuberculosis in South Africa, strenuous measures are needed to implement a good control programme that will increase the cure rate of tuberculosis patients. The results of this study suggest the need for health workers to learn about local beliefs that may influence presentation and adherence, and for traditional and western health workers to collaborate. PMID- 12546116 TI - Cost-effectiveness of different treatment strategies for tuberculosis in Egypt and Syria. AB - SETTING: The National Tuberculosis Programmes in Egypt and Syria. OBJECTIVES: To calculate the costs and effectiveness of alternative ways of implementing TB control in Egypt and Syria, in order to illustrate the factors influencing the cost-effectiveness of TB treatment in middle-income countries. DESIGN: We compared the costs and cure rates in Egypt and Syria of the World Health Organization recommended directly observed treatment, short-course (DOTS) strategy and alternative strategies. The study included costs both to the health services and to the patient. RESULTS: In Egypt and Syria, the cost-effectiveness of DOTS implemented through the primary health care (PHC) system was respectively $258 and $243 per patient cured. This compares to a cost per patient cured of $297 (Egypt) and $693 (Syria) for alternative strategies implemented through specialist clinics. In Egypt, when DOTS is implemented through specialist chest clinics it costs $585 per patient cured. Hospitalisation costs either $1490, $1621 or $1699 per patient cured, depending on treatment delivery in the continuation phase. CONCLUSION: This study demonstrates that the move towards DOTS integrated at the PHC level has substantially improved the effectiveness of TB treatment in Egypt and Syria, without substantially increasing costs. An analysis of the different costs and effectiveness of the variety of TB treatment strategies has enabled both National Tuberculosis Programmes to expand DOTS and implement it in a way that takes into account limited resources and local health systems. PMID- 12546117 TI - Effectiveness of tuberculosis control programmes in prisons, Barcelona 1987-2000. AB - SETTING: Different bodies have emphasised the need for prisons to implement and evaluate their own tuberculosis prevention and control programmes (TPCP), with the aims of evaluating TPCP in Barcelona prisons and obtaining conclusions that would allow any necessary modifications to be introduced to improve their effectiveness. METHODS: An assessment was carried out for the period 1987-2000, using the exhaustive retrospective method. The quantitative indicators used were defined in the same way as those employed by the Barcelona TPCP, which were taken as the gold standard. The assessment studied trends in the indicators and evaluated the impact by comparing the results with the gold standard. RESULTS: Statistically significant trends (P < 0.0001) were found for incidence rates, infection prevalence, treatment adherence rates among smear-positive cases and percentage of cases on directly observed treatment (DOT). When impact was assessed, only the decline in the average annual incidence rate (25.2%) and desired rates of adherence among smear-positives (> or = 95% since 1993) achieved the proposed objectives. CONCLUSIONS: The observed decline in incidence shows that when programmes are based on reducing diagnostic delay, controlling adherence using DOT and tracing contacts, tuberculosis control among the prisons is effective. PMID- 12546118 TI - Study of drug resistance in previously treated tuberculosis patients in Gujarat, India. AB - SETTING: Department of Tuberculosis and Chest Diseases and State Tuberculosis Diagnosis and Training Centre (STDTC), a DOTS centre in Ahmedabad, Gujarat State, India. The study was carried out by retrospectively reviewing patient data between January 2000 and August 2001. OBJECTIVE: To evaluate the pattern of drug resistance among previously treated tuberculosis patients who remained symptomatic or smear-positive despite receiving anti-tuberculosis drugs under DOTS for a minimum of 5 months. DESIGN: A total of 1472 pulmonary tuberculosis patients who had taken anti-tuberculosis treatment were evaluated retrospectively with respect to their drug resistance pattern by sputum culture for acid-fast bacilli (AFB) and sensitivity testing with isoniazid, rifampicin, streptomycin and ethambutol (E). RESULT: Of the 1472 patients evaluated, 804 (54.6%) were treatment failure cases and 668 (45.4%) were relapse cases; 822 patients (373 failure and 449 relapse) were culture-positive. Of these 822 patients, 482 (58.64%, 261 failure and 221 relapse) were resistant to one or more drugs. Resistance to one drug was observed in 86 patients (10.46%), to two drugs in 149 (18.13%), to three drugs in 122 (14.84%) and to four drugs in 125 (15.21%). Single drug resistance was most commonly seen with isoniazid (62 patients, 7.5%), followed by streptomycin (12 patients, 1.4%), rifampicin (eight patients, 0.97%) and ethambutol (four patients, 0.4%). Resistance to isoniazid plus rifampicin alone was seen in 76 patients (9.2%). CONCLUSION: Drug resistance is a major problem in the treatment of pulmonary tuberculosis. Detection of drug resistance patterns and treatment with second-line anti-tuberculosis drugs in appropriate regimens are necessary in the treatment of failure and relapse cases in order to reduce the emergence of multidrug-resistant tuberculosis. PMID- 12546119 TI - Results of a 12-month regimen for drug-resistant pulmonary tuberculosis. AB - SETTING: Several therapeutic regimens for drug-resistant tuberculosis have been suggested, most of them with a total duration of 18-24 months. OBJECTIVE: To report our experience using a shorter regimen. DESIGN: Fifty patients with drug resistant pulmonary tuberculosis were managed by withdrawing all anti tuberculosis drugs until the results of a drug sensitivity test were obtained (approximately 3 months), and then a 12-month self-administered regimen with four to six anti-tuberculosis drugs at full daily doses was initiated, based primarily on the sensitivity test and secondarily on the history of previous treatment. RESULTS: In 31 patients treatment was completed as planned, in six it was irregular and 13 definitively abandoned it. In the best scenario, 90.3% (28/31) of patients with full treatment were cured; this outcome was similar for both multidrug-resistant (MDR, n = 18, 88.9%) and non-MDR (n = 13, 92.3%) patients. Six months later, the relapse rate was 4.8%, and after a 5-year follow-up 14 out of 18 cured patients who were located remained asymptomatic (77.8%). If the worst scenario was applied, a 62.0% cure rate (31/50) was obtained. CONCLUSIONS: A 12 month regimen with a minimum of four anti-tuberculosis agents at full dose, essentially selected on drug sensitivity testing, could be an alternative option for the treatment of drug-resistant pulmonary tuberculosis. PMID- 12546120 TI - Adverse effects of BCG revaccination: a report on 13 cases from Rio de Janeiro, Brazil. AB - SETTING: BCG revaccination is not used worldwide. In Brazil, it has been recommended since 1994 in children aged 6 to 14 years. Reports in the medical literature of adverse reactions due to revaccination are rare. BCG revaccination remains officially recommended by the Brazilian health authorities, based on the current epidemiological tuberculosis situation in the country. OBJECTIVE: We report 13 cases of children and adolescents with complications due to BCG revaccination. DESIGN: Case reports of patients followed up in a pneumology unit of a university pediatric hospital from May 1996 to December 2000. RESULTS: There were seven males and six females, whose ages ranged from 7-12 years. Adverse reactions occurred between 21 days and 9 months after BCG revaccination. All 13 cases had a scar from prior BCG vaccination. All cases treated received isoniazid, 10 mg/kg/day, in the morning until the end of cure. The duration of treatment varied, but all cases were successfully cured. CONCLUSION: Although adverse reactions are rare, it is important to continue monitoring them in order to learn more about them and to be able to orient health professionals to perform appropriate assessment and timely treatment when they occur. PMID- 12546121 TI - A high tuberculosis case-fatality rate in a setting of effective tuberculosis control: implications for acceptable treatment success rates. AB - Worldwide, the case-fatality rate of smear-positive pulmonary tuberculosis among persons on treatment is 3.8%. We assessed the case-fatality rate among such patients in Baltimore between January 1993 and June 1998. Tuberculosis incidence was < 17/100 000 population, and 99% of patients received directly observed therapy. Of 174 patients, 42 (24%) died on treatment. Patients who died were older (mean age 62 vs. 47 years; P < 0.001) and were more likely to have underlying medical conditions. In multivariate analyses, older age, diabetes mellitus, and renal failure were independently associated with an increased risk of death. With effective control, tuberculosis may become concentrated in older persons with chronic diseases and be associated with high case-fatality rates. In such settings, acceptable treatment success rates may need to be revised. PMID- 12546122 TI - Rapid and simple MTT method for rifampicin and isoniazid susceptibility testing of Mycobacterium tuberculosis. AB - The MTT method for rifampicin and isoniazid susceptibility testing of Mycobacterium tuberculosis was developed by using bacterial suspension prepared from colonies on solid media. The MTT tube assay in 1 ml Middlebrook 7H9 broth was completed within 4 days for rifampicin (RMP) and within 7 days for isoniazid (INH). When MTT assay results with 279 M. tuberculosis clinical isolates were compared with those of the conventional proportion method on Lowenstein-Jensen medium, high specificity and sensitivity values of 100% and 94.1%, respectively, for RMP susceptibility testing, and 99.5% and 89.2%, respectively, for INH susceptibility testing were obtained. The accuracy of the MTT method for RMP and INH was > 0.97 concordance with the proportion method. The MTT method is simple, inexpensive and rapid. The high level of agreement with the conventional proportion method suggests a potential to rapidly detect drug-resistant M. tuberculosis in developing countries, as only basic microbiological equipment is need. PMID- 12546123 TI - [Smoking cessation program in Tunisia: experience of a respiratory service]. AB - A tobacco cessation programme was recently introduced into medical practice in Tunisia. The medical staff in the pulmonary disease ward at Charles Nicolle Hospital, Tunis, has been the first to implement such a programme for people motivated to quit smoking. This programme has been run for 3 years in the respiratory disease out-patient department. It acts essentially against psychological and pharmacological dependence on tobacco. The results obtained were very encouraging: 298 smokers have participated in this programme. The global success rate at 12 months of tobacco withdrawal was about 25% for people who were followed up, and 17% for the whole sample. Medical help for smoking cessation should be more widely promoted in Tunisia to increase the number of smokers who consult and improve the quality of medical intervention. PMID- 12546124 TI - Widespread distribution of a single drug rifampicin formulation of inferior bioavailability in South Africa. PMID- 12546125 TI - Phage tests for diagnosis and drug susceptibility testing. PMID- 12546126 TI - Use of the FASTPlaque test for TB diagnosis in low-income countries. PMID- 12546127 TI - Superficial fungal infection of the skin during treatment of tuberculosis. PMID- 12546128 TI - State and association/certifying boards: CE requirements. PMID- 12546129 TI - Certificate programs in continuing professional education. AB - BACKGROUND: Career-long learning is a concept at the heart of professional disciplines such as nursing. Nursing shortages, especially in some areas of practice, have stimulated the need for in-depth education and training in specific knowledge content areas. One approach to career-long learning is certificate programs. METHOD: Factors, trends, and strategies to consider in creating certificate programs are reviewed, and guidelines for developing certificate programs are presented. FINDINGS: Certificate programs are increasingly in demand by both employers and nurses. Certificate programs provide individuals with a chance to refresh their study skills and raise their abilities and levels of confidence for mastering new learning. Certificate programs also serve as a recruitment and retention strategy for hospitals and agencies. CONCLUSION: Certificate programs are a viable continuing professional education activity and offer opportunities for nurses to develop specific skill sets and in depth knowledge in specialized areas of nursing practice. PMID- 12546130 TI - The impact of graduate education on reading for lifelong learning. AB - BACKGROUND: Reading is one means of achieving professional lifelong learning, and while nursing as a profession has an abundance of literary resources, the amount of time nurses actually spend reading is unknown. Previous studies found nurses who had graduate degrees, were in leadership positions, and held multiple memberships in professional nursing organizations reported increased reading time. This study examined the amount of time RN students entering graduate school and graduate school alumni spend reading as well as their primary purpose (interest, application, and commitment) for the reading. METHOD: A descriptive/exploratory survey research design was used. The Reading Activity Inventory Tool was administered to new graduate (entry) students (n = 47) during orientation to their graduate program and graduate alumni (n = 99) who were 1, 2, and 3 years postcompletion of their masters degrees. Participants were recruited from four Texas nursing schools. FINDINGS: The average weekly time participants reported reading professional literature was 10 hours and 50 minutes for the entry group and 6 hours for the alumni group. As to the purpose (interest, application, and commitment) of reading the nursing literature, there was no significant difference on total interest or total application among the four cohorts (entry students and 1st-, 2nd-, and 3rd-year alumni). Entry students had a higher total commitment to reading than alumni nurses. CONCLUSION: While entry students spent more time reading in this study, the alumni with advanced degrees seemed invested in the profession with significantly more journal subscriptions, increased memberships in nursing organizations, and consistent reading activities. Strategies to encourage all nurses to read for lifelong learning need to be explored. PMID- 12546131 TI - The impact of education on the use of physical restraints in the acute care setting. AB - BACKGROUND: Physical restraints have become an acceptable standard of practice for managing safety and behavior control in acute care settings. Although the primary intent for using physical restraints is for patient protection, there are many negative outcomes related to their use. Heightened awareness by recent Joint Commission on Accreditation of Healthcare Organizations and the Center for Medicare and Medicaid Services standards for restraint use has led health care administrators and nursing staff to explore methods of reducing or eliminating the use of restraints. An educational program was planned and implemented for nursing staff emphasizing the risks of physical restraints and the benefits of innovative optional measures including nonrestraint devices. The program was tested to determine whether increased awareness through education would reduce the use of restraints. METHOD: After current practice patterns and restraint utilization were established, a comprehensive educational program was provided to all nursing personnel. Twenty-three formal classroom inservice offerings were provided with follow-up reinforcement of self-study modules. Education included nursing assessment strategies and practical restraint optional interventions for managing patients exhibiting disruptive behaviors. FINDINGS: After the educational program, the overall use of physical restraints decreased as well as the length of time patients were restrained. CONCLUSION: Results of this study reinforce the need to increase staff awareness and knowledge of nonrestraint interventions to manage disruptive behaviors in the acute care setting. PMID- 12546132 TI - Learning with cases. AB - BACKGROUND: The use of case studies as a teaching and learning method has been traced back to the Medical Society of New Haven in 1788. The use of case studies was introduced into Harvard Law School in 1871, and this method was used with the first class to graduate from Teachers College in 1930. Case studies are now widely used by other disciplines and are gaining popularity in nursing. METHOD: A review of the literature on the use of case studies for learning was performed. FINDINGS: Education is being transformed from a teaching to a learning focus. CONCLUSION: Case methods or studies provide a process of participatory learning that facilitates active and reflective learning and results in the development of critical thinking and effective problem-solving skills. This develops self directed lifelong learners. PMID- 12546133 TI - Professional development needs of registered nurses practicing at a military community hospital. AB - BACKGROUND: Focus groups were used to determine what factors need to be considered when designing a professional development program for RNs practicing at a military community hospital. METHOD: Six focus groups were formed with RNs from multiple directorates and settings within various specialties. FINDINGS: Five thematic groupings were identified: 1) specific professional development needs for leadership, clinical/specialty practice, competence development, and maintenance, 2) methods for providing continuing education, 3) methods for evaluating the effectiveness and efficiency of continuing education, 4) barriers to professional development, and 5) professional development issues impacting retention of military nurses. CONCLUSION: The professional development needs of RNs practicing in military hospitals are complex and multifaceted. The thematic groupings identified guided the design of a professional development program for RNs. PMID- 12546134 TI - Inhibition of prostaglandin G/H synthase unveils a potent effect of platelet activating factor on the permeability of bovine aortic endothelial cells to albumin. AB - Platelet Activating Factor (PAF) is a very potent stimulant of various cell functions but little is known about the mechanisms responsible for its marked effect on endothelial permeability. An in vitro assay system was used to assess the direct effect of PAF on the permeability of a bovine aortic endothelial cell (BAEC) monolayer to albumin. PAF produced a small but not significant increase of the permeability of BAEC monolayer to albumin. However, pre-treatment of the monolayer with indomethacin (10 microM) resulted in a significant increase of BAEC permeability following PAF administration. This increase was concentration dependent up to a maximal effect of 105% above basal value (for 0.1 microM PAF). Addition of the PAF antagonist SRI 63 441ZI (5 microM) abolished this effect. Exogenous administration of PGE2 (10(-7) M) inhibited the effect of PAF on the BAEC permeability suggesting that prostaglandins synthesized by the endothelium behave as a negative autoregulatory factor. Compound SRI 63 441ZI also partially inhibited bradykinin-induced permeability to albumin but did not significantly modify the activity of thrombin. These findings show that PAF can increase endothelial permeability to albumin when the synthesis of prostaglandins is inhibited. Our results also show that PAF might have an autocrine activity by mediating part of BK-induced permeability. PMID- 12546135 TI - Morphine-induced enhancement in the granulocyte response to thioglycollate administration in the rat. AB - The present study determined the pharmacological effects of acute morphine treatment on the granulocyte phase of the peritoneal inflammatory response to thioglycollate (TG) in rats. Dual-color flow cytometry using anti-CD11b/c-PE mAb in combination with HIS48-FITC mAb allowed for the determination of morphine's effects on 4 inflammatory cell subsets: CD11b/c(+)HIS48med granulocytes; CD11b/c(hi)HIS48neg/lo activated macrophages; CD11b/c(-)HIS48(-) lymphocytes; and CD11b/c(+)HIS48hi cells (a monocyte/macrophage and granulocyte subset). Morphine produced a dose-dependent increase in a select subset of inflammatory peritoneal cells, the CD11b/c(+)HIS48med granulocytes. The effect of morphine was time dependent, with significant effects first apparent at 4 hr after TG, but the administration of morphine 1 hr before or simultaneously with TG produced a similar increase in CD11b/c(+)HIS48med granulocytes. Naltrexone completely antagonized the morphine-induced increase in CD11b/c(+)HIS48med granulocytes. Collectively, these studies show that a single administration of morphine produces a time-dependent, dose-dependent, opioid receptor-mediated enhancement in the peritoneal granulocyte response to TG. PMID- 12546136 TI - NADPH oxidase priming and p47phox phosphorylation in neutrophils from synovial fluid of patients with rheumatoid arthritis and spondylarthropathy. AB - Superoxide anion (O2(o)-)production by neutrophil NADPH oxidase participates in arthritic joint lesion formation. Proinflammatory cytokines such as tumor necrosis factor alpha (TNFalpha), interleukin 8 (IL-8) and granulocyte/macrophage colony stimulating factor (GM-CSF) have a priming effect on neutrophil NADPH oxidase activity. NADPH oxidase activation is dependent on phosphorylation of p47phox, a cytosolic component of the enzyme. We studied O2(o)-production and p47phox phosphorylation in synovial fluid (SF) from patients with rheumatoid arthritis (RA) and spondylarthropathy (SpA) according to TNFalpha, IL-8 and GM CSF levels. O2(o)-production by neutrophils isolated from SF of all the arthritis patients (RA and SpA) was higher than that of circulating resting neutrophils and when stimulated with fMLP or PMA. In addition, p47phox was partially phosphorylated in SF neutrophils compared to circulating neutrophils. High levels of TNFalpha and IL-8 (but not GM-CSF) are detected in patient's SF (compared to circulating blood levels). TNFalpha levels were significantly higher in RA than in SpA SF. These results suggest that increased NADPH oxidase activity could be involved in arthritic joint inflammation through increased p47phox phosphorylation. This could be the result of the presence of high levels of priming agents such as TNFalpha and IL-8 but not GM-CSF. PMID- 12546137 TI - Glucocorticoids modulate TGF-beta production. AB - TGF-beta is thought to play a central role in pulmonary fibrosis inducing fibroblast differentiation and extracellular matrix synthesis. In human lung fibroblasts, it is still unclear how various TGB-beta isoforms affect TGF-beta production and whether glucocorticoids, commonly used agents to treat fibrotic lung disease, modulate these processes. To this end, human fetal lung fibroblasts (HFL-1) were cultured with various concentrations of glucocorticoids (budesonide, dexamethasone or hydrocortisone) with and without TFG-beta1, -beta2, and -beta3. TGF-beta mRNA was assessed by real time RT-PCR. Smad 2, 3, and 4 and AP-1 complex (c-fos and c-Jun) cellular localization were evaluated by immunostaining. TGF beta2 and -beta3 stimulated TGF-beta1 production significantly (p < 0.01 relative to control). TGF-beta1 stimulated TGF-beta2 production (p < 0.01 relative to control). TGF-beta3 was undetectable. Glucocorticoids significantly inhibited TGF beta1 and -beta2 production and reduced expression of the upregulated TGF-beta1 and -beta2 mRNA induced by exogenous TGF-beta1, -beta2 or -beta3 (p < 0.01 for each) but had no effect on Smads. Although c-jun-related nuclear staining was not intensified in TGF-beta-stimulated cells, it was reduced by glucocorticoids. Thus, TGF-beta isoforms may stimulate production of various TGF-beta isoforms in the lung. Glucocorticoids then may block TGF-beta production by modulating mRNA levels and c-Jun. PMID- 12546138 TI - Modulation of plasma levels of soluble adhesion molecules and nitric oxide in healthy volunteers by exposure to swine dust. AB - Short-time exposure to swine dust causes an intense airways inflammation and symptoms of systemic inflammation in healthy volunteers. Here, we sought to study whether this response involved signs of endothelial cell activation. Peripheral blood cell counts and plasma levels of sE-selectin, sP-selectin, sICAM-1, interleukin-8, nitrite and nitrate were measured in blood samples from 17 healthy subjects before and after a 3-hr exposure to swine dust in a swine confinement building. Dust exposure induced a 3-fold increase of blood neutrophil p = 0.0009) and 1.5-fold increase of monocyte counts (p = 0.0047). IL-8 was detected in 15 individuals after exposure (p = 0.001). Endothelial cell markers such as sICAM and nitrate increased by 10 and 34% resp. (p = 0.011 and 0.017), whereas sE selectin remained unchanged and sP-selectin was reduced by 15% (p = 0.031). Thus, short time exposure to swine dust induced a systemic inflammatory response with evidence of endothelial and inflammatory cell activation. PMID- 12546139 TI - Tissue-specific ICAM-1 expression and neutrophil transmigration in the copper deficient rat. AB - Dietary copper deficiency promotes neutrophil accumulation in rat lungs. We have now investigated the potential mechanisms of this effect. Male weanling rats were fed a Cu-adequate (6.0 mg diet) or Cu-deficient diet (0.30 mg) for 4 wks. Endothelial intercellular adhesion molecule-1 (ICAM-1) expression was measured in vivo and in vitro using a radiolabeled monoclonal antibody to rat ICAM-1. Tissue neutrophil accumulation was measured by myeloperoxidase (MPO) content and neutrophil transendothelial migration was assessed in vitro. Dietary copper deficiency had no effects on the expression of ICAM-1 in lung, liver, heart, kidney, or cremaster. However, MPO content was significantly greater in the lungs of copper-deficient rats. Endotoxin-induced ICAM-1 expression was greater in the lungs and hearts of copper-deficient rats. Similarly, cultured rat endothelial cells that were Cu-chelated expressed more ICAM-1 after endotoxin. This correlated with the significant increase in MPO in lungs of copper-deficient rats treated with endotoxin. The results suggest a tissue-specific difference in ICAM 1 expression and neutrophil accumulation during inflammation in copper-deficient rats. The findings suggest that lung inflammatory mechanisms are particularly sensitive to copper deficiency. PMID- 12546140 TI - Differential effect of Fc gamma receptor ligation on LPS-stimulated TNF-alpha secretion by hepatic, splenic, and peritoneal macrophages. AB - Lipopolysaccharide (LPS) increases serum TNF-alpha levels due to TNF-alpha secretion by macrophages. The serum TNF-alpha response to LPS was augmented 10x when FcgammaR ligation was induced by the intravenous injection of Gig-coated erythrocytes (IgG) prior to the administration of LPS. The macrophage population responsible for the augmented TNF-alpha secretion was determined by isolating Kupffer cells, splenic macrophages and peritoneal macrophages from mice that had been given ElgG prior to LPS and determining TNF-alpha secretion ex vivo. The intravenous injection of ElgG augmented LPS-stimulate TNF-alpha secretion by Kupffer cells and splenic macrophages. In contrast, LPS-stimulated TNF-alpha secretion by peritoneal macrophages was not altered by either the intravenous or intraperitoneal injection of ElgG. In vitro phagocytosis of ElgG by isolated peritoneal macrophages also did not augment LPS-stimulated TNF-alpha secretion. These results show that FcgammaR ligation augments LPS-stimulated TNF-alpha secretion by Kupffer cells and splenic macrophages but not by peritoneal macrophages. Thus, the ability of FcgammaR ligation to influence TNF-alpha secretion may be specific to the tissue source of the macrophages. PMID- 12546141 TI - Modulation of leukocyte recruitment and IL-8 expression by the membrane attack complex of complement (C5b-9) in a rabbit model of antigen-induced arthritis. AB - The complement system is thought to be a major physiological mediator of injury in a number of diseases including rheumatoid arthritis (RA). The membrane attack complex (MAC) of complement has been detected in RA tissue, suggesting that the MAC may be relevant to the pathogenesis of the disease. Deposition of sublytic concentrations of the MAC has been shown to promote the expression of proinflammatory mediators. In the present study, we utilized rabbits deficient in the complement protein C6 to elucidate the role of the MAC in mediating the pathogenesis of antigen-induced arthritis. Swelling, leukocyte accumulation, IL-8 expression, proteoglycan, and hydroxyproline content were assessed. Analysis of synovial tissue demonstrated a significant decrease in leukocyte influx and a parallel decrease in tissue associated IL-8 in joints of C6-deficient animals as compared to C6-sufficient animals. However, this did not correlate with the preservation of connective tissue. The results derived from this study provide evidence that the MAC has an important function in mediating leukocyte recruitment in antigen-induced arthritis but does not play a direct role in connective tissue breakdown. PMID- 12546142 TI - Optimization of single-phase, unidirectional transducers using three fingers per period. AB - Electrode width controlled (EWC) single-phase, unidirectional transducers (SPUDT) is widely used for low loss surface acoustic wave (SAW) filters. The insertion loss and fractional bandwidth of the filters are strongly related to the reflectivity of EWC cells. In order to achieve wide band and low loss simultaneously, it is necessary to obtain higher reflectivity. The relationship between geometrical configuration of EWC cells and reflection coefficient, (and transduction coefficient as well) is studied. Simulation results indicate that the reflectivity of the EWC SPUDT cell could exceed 5% on a 128 degrees Y-X lithium niobate (LiNbO3) substrate. Using such structure, low loss SPUDT test filters without weighting are fabricated. The measured 3 dB bandwidth is 3.9% and the insertion loss is 2.9 dB. The theoretical calculation is verified by the experiment. PMID- 12546143 TI - Anisotropy in domain engineered 0.92Pb(Zn1/3 Nb2/3)O3-0.08PbTiO3 single crystal and analysis of its property fluctuations. AB - The orientation dependence of slowness and electromechanical coupling coefficients of 0.92Pb(Zn1/3 Nb2/3)O3-0.08PbTiO3 (PZN-8%PT) domain engineered single crystal was analyzed based on the measured complete set of elastic, piezoelectric, and dielectric constants. There exist one quasi-longitudinal, one quasi-shear, and one pure shear wave in each of the [100]-[010], [010]-[001], and [001]-[110] planes. The slowness of the quasi-shear wave exhibits strong anisotropy in all three planes, and the coupling coefficient k33 and k31 reach their maximum in [001] and [110] directions of cubic axis, respectively. Because the composition of PMN-8%PT system is very close to the morphotropic phase boundary, the extraordinary large piezoelectric coefficients d31 and d33, and high coupling coefficient k33 are very sensitive to compositional variation. We have performed error analysis and proposed an improved characterization scheme to derive a complete data set with best consistency. PMID- 12546144 TI - High Q-factor Bragg-reflection sapphire-loaded cavity TE01delta mode resonators. AB - An innovative method of enhancing the quality factor of TE01delta cavity resonators with a dielectric tube made of monolithic sapphire is presented. Very high Q-factor is achieved by employing a Bragg reflection technique. A TE01delta mode in a copper cavity was measured to have a Q-factor of 1 x 10(5) at 8.78 GHz and 290 K. This is only 30% less than the limit due to the loss tangent of the dielectric material. The technique confines electromagnetic energy in the sapphire dielectric and in the vacuum well away from the cavity walls, thus reducing the surface losses in the copper shield. The technique offers some significant advantages over other methods. One advantage is the very low spurious mode density, which can improve filter and resonator design capabilities. Another is the small compact design, with a single sapphire piece, as compared to previously published Bragg reflection techniques. Finite element simulations and experimental data for this method were compared and found to be in very good agreement. The cavity dimensions were optimized to achieve maximum quality factor. PMID- 12546145 TI - Monolithic bulk shear-wave acousto-optic tunable filter. AB - We demonstrate a monolithic bulk shear-wave acousto-optic tunable filter combining a piezoelectric transducer array and the acoustic interaction medium in a single crystal. An X-propagating acoustic longitudinal wave is excited in the "crossed-field" scheme by an rf-Ey-field in a chirped acoustic superlattice formed by domain-inversion in lithium niobate (LiNbO3). The acoustic longitudinal wave is efficiently (97.5%) converted at a mechanically free boundary into a Y propagating acoustic slow-shear wave that couples collinearly propagating e- and o-polarized optical waves. A relative conversion efficiency of 80%/W was measured at 980 nm. PMID- 12546146 TI - Optical and acoustical interrogation of submicron contrast agents. AB - Unlike conventional ultrasound contrast agents with a diameter of several microns, in this paper we explore the use of submicron contrast agents for the detection and localization of lymph nodes. The submicron agents are gas-filled, double-walled microspheres that rupture when exposed to ultrasound energy at megahertz frequencies. In this study, three experimental systems are combined with model predictions to assist in understanding the response of these unique agents to a range of signal transmission parameters. Optical experimental results for each agent delineate the relative expansion as a function of acoustical peak negative pressure, pulse length, and center frequency. The optical images demonstrate an order of magnitude expansion in radius during the pulse rarefaction, in which the expansion magnitude is dependent on the transmitted pressure and frequency. Simulations using a modified Rayleigh-Plesset model predict an increasing relative expansion for the microbubbles (initial bubble radius ranging from 0.3-1.3 microm) with increasing pressure and decreasing initial radius. Acoustically recorded frequency spectra reveal the presence of harmonics for a range of transmitted pulses. In addition, in-vivo results from a normal canine model demonstrate marked contrast enhancement of first order lymph nodes. We hope to offer an alternative to present intra-operative procedures for sentinel node detection. PMID- 12546147 TI - Optimizing the beam pattern of a forward-viewing ring-annular ultrasound array for intravascular imaging. AB - Intravascular ultrasound (IVUS) imaging systems using circumferential arrays mounted on cardiac catheter tips fire beams orthogonal to the principal axis of the catheter. The system produces high resolution cross-sectional images but must be guided by conventional angioscopy. A real-time forward-viewing array, integrated into the same catheter, could greatly reduce radiation exposure by decreasing angiographic guidance. Unfortunately, the mounting requirement of a catheter guide wire prohibits a full-disk imaging aperture. Given only an annulus of array elements, prior theoretical investigations have only considered a circular ring of point transceivers and focusing strategies using all elements in the highly dense array, both impractical assumptions. In this paper, we consider a practical array geometry and signal processing architecture for a forward viewing IVUS system. Our specific design uses a total of 210 transceiver firings with synthetic reconstruction for a given 3-D image frame. Simulation results demonstrate this design can achieve side-lobes under -40 dB for on-axis situations and under -30 dB for steering to the edge of a 80 degrees cone. PMID- 12546148 TI - Approach to 3-d ultrasound high resolution imaging for mechanically moving large aperture transducer based upon Fourier transform. AB - A new three-dimensional (3-D) acoustic image formation technique is proposed that is based on the transmission of wide bandwidth pulse signals and the application of the 3-D fast Fourier transform. A solution to the Helmholz wave equation has been obtained using the Born approximation. The solution contains analytical expressions for the spatial spectra of the transmit and receive radiation patterns for transducers of various geometries with lenses of fixed focal distances. It has been shown that the proposed algorithms allow for radiation patterns with constant widths at depths both behind and in front of the focal point, starting practically from the plane of the transducer. The theoretical and experimental investigations and computer simulation for both spherical and rectangular transducer shapes have been performed. The results were used to estimate the beamwidths and the side lobe levels. A variant of the linear array has been studied for cylindrical lens of a fixed focal distance moving in a lateral direction. It has been shown that, in this case, a high resolution (of the order of a few wavelengths) can be achieved along all three Cartesian coordinates at a very high scanning velocity. The influence of the moving scatterers' velocity in inhomogeneous medium on the spatial radiation pattern characteristics has been estimated. PMID- 12546149 TI - Analysis and correction of ultrasonic wavefront distortion based on a multilayer phase-screen model. AB - A model is introduced that incorporates the cumulative wavefront distortion effects caused by spatial heterogeneities along the path of propagation, and a corresponding model-based wavefront distortion-correction method is presented. In the proposed model, a distributed heterogeneous medium is lumped into a series of parallel phase screens. The distortion effects can be compensated--without a priori knowledge of the distorting structure--by backpropagation of received wavefronts through hypothetical multiple phase screens located between the imaging system and targets, while each pointwise time shift is adjusted iteratively to maximize a specified image quality factor at the final layer. Theoretical analyses indicate that the mean speckle brightness decreases monotonically with the root-mean-square value of distributed phase distortions; therefore, the speckle brightness can be used as an image quality factor. Experimental one-dimensional (1-D) array data with simulated distortion effects based on a real 2-D abdominal-tissue map were used to evaluate the performance of the proposed method and existing aberration-correction techniques. The simulated characteristics of wavefront distortion and relative performance of existing correction techniques were similar to reports based on abdominal-wall data and breast data. This investigation shows that the proposed method provides better compensation for wavefront distortion. PMID- 12546151 TI - Angular-profile tuning of guided waves in hollow cylinders using a circumferential phased array. AB - Angular-profile tuning of guided waves in hollow cylinders is implemented by using partial loading of the elements in a circumferentially placed phased array. Each partial loading element generates nonaxisymmetric guided waves in a pipe. In earlier work, numerical calculations and experiments have shown that, for nonaxisymmetric guided waves, circumferential distribution of particle displacements (i.e., the angular profile) changes with propagation distance, frequency, and mode. To change the angular profile at a certain distance, either frequency or mode has to be changed for a single partial loading element. This is not the case, however, for a circumferential phased array. The total angular profile of a circumferential array is the superposition of contributions from all elements. If given the knowledge of the angular profile for a single element, the total guided wave angular profile can be controlled and thus focused at any specific circumferential location by a circumferentially placed phased array with adjustable voltage level and phase inputs. This angular profile tuning technique can be used for implementing a circumferential scan with focused, guided wave beams, which leads to the detection of smaller defects as a result of stronger focused beams. Algorithms and specific nondestructive evaluation (NDE) applications for pipe inspection using this technique are discussed. PMID- 12546150 TI - Processing radio frequency ultrasound images: a robust method for local spectral features estimation by a spatially constrained parametric approach. AB - Spectral estimation is a major component in studies aiming at characterizing biological tissues through the analysis of backscattered radio frequency (RF) ultrasonic signals and images. However, conventional spectral estimation techniques yield a well-known trade-off between spatial resolution and variance. The backscattered signals are stochastic by nature, so short-term local analysis results in a high variance of the estimates, which cannot efficiently be reduced through conventional spatial averaging. We address this issue by describing a spectral estimation technique that reduces the variance of the estimates (by smoothing the local estimates in spectrally homogeneous regions) while preserving spectral discontinuities (i.e., the smoothing is not performed across regions with different spectral contents). The proposed approach is set in a Bayesian framework and is based on local autoregressive (AR) estimation, constrained by smoothness priors. These smoothness priors are introduced through a Markov random field in which the associated potential functions are nonquadratic, allowing thereby to preserve discontinuity. The method is validated on simulated RF images and tested on echocardiographic images acquired in vivo. The results are compared to the estimates provided by the conventional Burg technique. These results clearly demonstrate the ability of the proposed approach to improve spectral estimation in terms of variance reduction and discontinuity detection. PMID- 12546152 TI - Reflection of plate acoustic waves produced by a periodic array of mechanical load strips or grooves. AB - The reflection of fundamental acoustic waves propagating in a thin piezoelectric plate by a periodic array of conducting strips of finite thickness or grooves has been theoretically and experimentally investigated. The analysis has shown that electrical shorting and mass loading affect the relationship of neighboring region impedances in a contrary manner. In some cases, these effects are comparable, and there exists a certain strip thickness for each piezoactive fundamental plate mode at which the reflection coefficient can become zero. A high efficiency of grooved reflector for plate acoustic waves was theoretically revealed. Experimental results for mass loading and grooved reflections, which have been obtained for an SH0 wave propagating in the Y-X lithium niobate plate, are in a good agreement with the theory. They show a high efficiency of such reflectors and confirm an opportunity of using a model based on equivalent circuit for the analysis of their operation. Investigations indicate that nearly 100% reflection of the SH0 wave in the lithium niobate plate can be obtained with the use of a mass loading reflector containing 10 silver strips of thickness d/h = 0.08 or a grooved reflector containing eight grooves of depth d/h = 0.25. Here h is the plate thickness and d is the reflector thickness or depth. PMID- 12546153 TI - Fast, precise, and full extraction of the COM parameters for multielectrode-type gratings by periodic Green's function method. AB - It is very important to extract all four coupling-of-modes (COM) parameters of the electrode cells for the simulation and optimal design of a low loss surface acoustic wave (SAW) filter. A new approach for fast and full extraction of the COM parameters for multielectrode-type grating is proposed. The field distribution of the wave under the periodic shorted grating is calculated by periodic Green's function method. The phase of the reflection is determined from the positions of the standing wave node. The transduction coefficient and its phase are determined by the charge distribution at low frequency. The COM parameters of the commonly used electrode width controlled (EWC) single phase unidirectional transducer (SPUDT) is computed. It shows that this is a simple and direct way to extract all the COM parameters for SPUDT and, accordingly, is a powerful tool for the optimization of the filter structure. PMID- 12546154 TI - A new high resolution color flow system using an eigendecomposition-based adaptive filter for clutter rejection. AB - We present a new signal processing strategy for high frequency color flow mapping in moving tissue environments. A new application of an eigendecomposition-based clutter rejection filter is presented with modifications to deal with high blood to-clutter ratios (BCR). Additionally, a new method for correcting blood velocity estimates with an estimated tissue motion profile is detailed. The performance of the clutter filter and velocity estimation strategies is quantified using a new swept-scan signal model. In vivo color flow images are presented to illustrate the potential of the system for mapping blood flow in the microcirculation with external tissue motion. PMID- 12546155 TI - Dr. Andras Guttman. PMID- 12546156 TI - Use of germanate as a background electrolyte for capillary zone electrophoresis. AB - The possibilities of using germanate as a background electrolyte (BGE) in capillary zone electrophoresis (CZE) were investigated. Germanate can be obtained with spectral purity and, due to its two pKa values, possesses higher buffer capacity and can be used in a wider pH range than borate. The advantages and disadvantages of germanate among other polyanions for use as a background electrolyte for CE analysis are discussed. PMID- 12546157 TI - Capillary electrophoresis with laser-induced fluorescence detection of indigo carmine and indigo carmine-labeled proteins. AB - Indigo carmine blue is a water-soluble, light-sensitive, anionic dye most widely applied in microscopic staining techniques. It is virtually nonfluorescent in its oxidized state in aqueous solution, but when reduced under alkaline conditions, it becomes fluorescent, with absorption and emission maxima at 436 and 528 nm, respectively. It is demonstrated that the fluorescent character of the reduced form of the dye can be exploited as a label for the determination of cationic proteins by capillary electrophoresis with laser-induced fluorescence detection. Model proteins trypsinogen and cytochrome c are noncovalently labeled with indigo carmine; the bound species are reduced, rendering the indigo carmine fluorescent; and capillary electrophoresis with laser-induced fluorescence detection is used for subsequent analysis. Various buffer systems and buffer additives were examined and optimized, and a suitable pH range for optimal fluorescence intensity and protein-dye interaction was established. Fluorescence quenching of the reduced dye when bound to protein was observed in all buffer systems. PMID- 12546158 TI - Determination of lamivudine/didanosine/nevirapine in human serum using capillary zone electrophoresis. AB - A combination of the anti-HIV drugs lamivudine (3TC), didanosine (ddI), and nevirapine were separated and quantitated in human serum with capillary zone electrophoresis (CZE). The effects of various factors such as run buffer concentration and pH on the separation were investigated. The optimized resolution was achieved with a run buffer containing 100 nM N,N dimethyloctylamine in 80 mM phosphate buffer (pH 2.5). Diltiazem was chosen as the internal standard. All analytes were separated within 10 min at 30 degrees C with a voltage of +20 kV and UV detection at 210 nm. PMID- 12546159 TI - Determination of physostigmine and pyridostigmine in pharmaceutical formulations by capillary electrophoresis. AB - Physostigmine (PHY) and pyridostigmine (PYR) are two important anticholinesterase compounds with several clinical uses. Recently, PHY has been investigated for the treatment of senile dementia in Alzheimer's disease. However, both PHY and PYR have gained importance as antidotes for anticholinergic drugs. In military medicine, PYR is used as a prophylactic against nerve gas poisoning and was used in Saudi Arabia during the Gulf War in 1991. A new capillary zone electrophoresis (CZE) method for the rapid determination of PHY and PYR in pharmaceutical preparations has been developed. An untreated fused-silica capillary tube (75 microm i.d., 44 cm total length, 36.5 cm length to the detector) was employed with detection at 200 and 270 nm for PHY and PYR, respectively. The optimal separation conditions for PHY were: 50 mM boric acid-HCl buffer (pH 3.25) with 30 mM NaClO4, electrokinetic injection for 5 sec at -5 kV, temperature 25 degrees C, and separation voltage 15 kV. The optimal separation conditions for PYR were: 20 mM phosphate buffer (pH 7), electrokinetic injection for 20 sec at -10 kV, temperature 25 degrees C, and separation voltage 15 kV. The limits of detection (LOD, S/N = 3) were 70 and 60 ppb for PHY and PYR, respectively. The method can be used for the monitoring of possible main degradation products in tablets of military antidote formulations. PMID- 12546160 TI - Validation of a capillary electrophoresis assay for assessing the metabolic stability of verapamil in human liver microsomes. AB - A simple CE assay for the rapid determination of the in vitro metabolic stability of verapamil in human liver microsomes has been developed and validated. Verapamil was used as the prototype drug since it is extensively metabolized in human liver microsomes. The assay showed good intra- (CV < or = 10%) and interday (CV < or = 8%) reproducibility. The recovery of verapamil after incubation at 37 degrees C for 60 min with human liver microsomes was low (15 +/- 1%) and two metabolites were detected. The method is currently in use for assessing the metabolic stability of new drug candidates at an early stage of lead optimization at Cardiome Pharma Corp. (Vancouver, BC, Canada). PMID- 12546161 TI - Electrophoretic mobilities and migrating analytes: Part 1: Relationships. AB - The molecular radii (r) of a series of peptides have been determined by molecular modeling. With these data, it is shown that electrophoretic mobility (mu(ep)) is proportional to 1/r2, and that the dependence presented in textbooks (mu(ep) infinity 1/r) is wrong. Use of the approximately equivalent, mass-based Offord equation is discussed, and other relevant considerations are presented. PMID- 12546162 TI - Electrophoretic mobilities and migrating analytes: Part 2: Hydration. AB - A molecular modeling package has been used to systematically hydrate a series of previously built peptides. The volume data derived have been used with the log log form of the inverse square law and published electrophoretic mobilities to test for the most likely states of hydration. Starting with the bare peptides and increasing hydration, however, the strength of correlation diminishes, indicating that the average electromigrating peptide is not extensively hydrated. The shapes of the peptides are predominantly oblate ellipsoidal, but generally do not deviate significantly from spherical and progressive hydration increases this trend. Thus, shape correction does not play a significant role in the calculations. PMID- 12546163 TI - Vulnerable women and neo-liberal globalization: debt burdens undermine women's health in the global South. AB - Contemporary processes of globalization have been accompanied by a serious deterioration in the health of many women across the world. Particularly disturbing is the drastic decline in the health status of many women in the global South, as well as some women in the global North. This paper argues that the health vulnerability of women in the global South is inseparable from their political and economic vulnerability. More specifically, it links the deteriorating health of many Southern women with the neo-liberal economic policies that characterize contemporary economic globalization and argues that this structure is sustained by the heavy burden of debt repayments imposed on many Southern countries. In conclusion, it argues that many Southern debt obligations are not morally binding because they are not democratically legitimate. PMID- 12546164 TI - Caring, social policy, and homelessness. AB - Care theory offers a way to overcome a weakness of liberalism--its reluctance to intervene in the private lives of adults. In caring for the homeless, we must sometimes use a limited form of coercion, but our intervention is always interactive, and the process of finding a solution is one of negotiation between the needs expressed by the homeless and the needs we infer for them. PMID- 12546165 TI - On being poor and feeling poor: low socioeconomic status and the moral self. AB - Persons of low socioeconomic status generally experience worse health and shorter lives than their better off counterparts. They also suffer a greater incidence of adverse psychosocial characteristics, such as low self-esteem, self-efficacy, and self-mastery and increased cynicism and hostility. These population data suggest another category of harm to persons: diminished moral agency. Chronic socioeconomic deprivation can create environments that undermine the development of self and capacities constitutive to moral agency--i.e., the capacity for self determination and crafting a life of one's own. The harm affects not only the choices a person makes, but the chooser herself. This moral harm is particularly salient in modern Western societies, especially in the United States, where success and failure is attributed to the individual, with little notice of the larger social and political realities that inform an individual's circumstances and choices. PMID- 12546166 TI - Hotep's story: exploring the wounds of health vulnerability in the US. AB - A wide variety of forms of domination has resulted in a highly heterogeneous health risk category, "the vulnerable." The study of health inequities sheds light on forces that generate, sustain, and alter vulnerabilities to illness, injury, suffering and death. This paper analyzes the case of a high-risk teen from a Boston ghetto that illuminates intersections between "race" and class in the construction of vulnerability in the US. Exploration of his "wounds" helps specify how large-scale social and cultural forces become embodied as individual experience of disparate health risk. The case demonstrates that health inequities would not occur if resources--employment, income, wealth, education, housing, profiling in the legal system, and health care--were more justly managed in keeping with standards outlined in the Universal Declaration of Human Rights. Professional responses to the "wounds of vulnerability" may reveal important aspects of who we are and what our work as scholars, practitioners, and advocates must become. PMID- 12546167 TI - Race and healthcare disparities: overcoming vulnerablity. AB - The paper summarizes recently published data and recommendations about healthcare disparities experienced by African Americans who have Medicare or other healthcare coverage. Against this background the paper addresses the ethics of such disparities and how disadvantages of vulnerable populations like African Americans are typically maintained in decision making about how to respond to such disparities. Considering how to respond to disparities reveals much that vulnerable populations would bring to the policy-making table, if they can also be heard when they get there. The paper argues that vulnerable populations like African Americans need fair representation in bodies deciding what to do about such disparities and that fairness requires proportional representation at all levels of decisions that affect healthcare--a radical change. In this decision setting, how to provide adequate protection of minorities needs much further attention. The most attractive decision-making model is deliberative democracy. The paper shows that in deliberation, fair representation requires not only having a voice in decisions, but a fair hearing of those voices. Achieving a fair hearing requires changes in norms of communication and training of all to give importance to greetings and other measures of civility and trust building, and to be open to diverse forms of expression. Decisions about how to respond to healthcare disparities would include what programs to initiate for whom, how to evaluate the programs, and what to do in response to such evaluations. Conclusions are that achieving such goals will take a sea change in how healthcare institutions and providers do their business, and that social activism at every level will be needed to effect these changes. The discussion highlights many ethical issues that need much greater attention. PMID- 12546168 TI - Confining choices: should inmates' participation in research be limited? AB - Historically, prisoners in the United States have served as an inexpensive and readily available source of human subjects for research. Coinciding with the civil rights movement, however, was an emerging conception of prisoners' rights that led to the National Commission for the Protection of Human Subjects of Biomedical and Behavioral Research being charged with investigating the use of prisoners as research subjects. The recommendations that evolved and the subsequent guidelines that have been implemented by the Department of Health and Human Services significantly curtail the use of prisoners as research subjects. While these measures are designed to protect inmates from the abuses of the past, of particular concern to many health care officials is exclusion of inmates from experimental HIV/AIDS and hepatitis treatments. This paper addresses whether the vulnerability of prisoners in the United States due to their incarceration is sufficient to prohibit them from participation in clinical trials that offer the possibility of life-saving treatment. It first outlines the evolution in moral thinking that has led to laws broadly prohibiting prisoners from biomedical research studies and then analyzes cases in the law to develop ethical arguments in support of the view that prisoners should be allowed to participate in clinical trials. The conclusion is that prisoners should be allowed to participate in such trials. PMID- 12546169 TI - Assessment of renal and hepatic hemodynamics in cirrhosis of liver. AB - BACKGROUND AND OBJECTIVE: Investigators have documented an increase in renal artery resistance in cirrhotic patients. We studied the hepatic and renal hemodynamics by color Doppler in patients with liver cirrhosis in relation to the presence and severity of ascites. METHODS: A prospective study was carried out in a large public hospital on 33 patients with cirrhosis of liver and 11 healthy control subjects. Eleven patients had no ascites, 12 had diuretic-responsive ascites and 10 had refractory ascites. The resistance index (RI) of the hepatic artery was measured by color Doppler on its main branch and renal artery RI was measured on the main renal, interlobar and cortical arteries of the right kidney. RESULTS: Considering intraobserver variabilities, coefficient of variation for RI was 2.3%-7.8% (mean [SD] 4.2% [1.8%]). Hepatic artery RI were 0.68 (0.07), 0.68 (0.1), 0.81 (0.04), 1.0 (0.12) in control subjects and cirhotics with no ascites, responsive ascites and refractory ascites, respectively. Renal RI was significantly higher than in control subjects at all three levels in patients with responsive and refractory ascites, being higher in those with refractory than responsive ascites. Renal RI decreased from hilum towards the outer parenchyma, in cirrhotics with no ascites and responsive ascites but not in refractory ascites. CONCLUSION: Hepatic artery RI is higher in cirrhotics and is related to presence and severity of ascites. In control subjects and patients with no ascites or responsive ascites the renal RI decreased from hilum to the outer parenchyma. PMID- 12546170 TI - Effect of omeprazole on plasma zinc levels after oral zinc administration. AB - BACKGROUND: The intestines are the major site of zinc absorption and excretion. Reduced gastric acid secretion and elevated gastric pH is an important factor affecting intestinal mineral absorption. METHODS: Gastric pH and volume, and basal and maximal acid outputs were measured in 14 healthy volunteers. Plasma zinc levels were then measured at baseline and 1, 2, 3 and 4 hours after oral administration of 300 mg zinc sulfate. The experiment was repeated after omeprazole administration (60 mg/day orally) for 7 days. RESULTS: Omeprazole administration significantly increased fasting gastric pH (5.5 versus 2.4; p < 0.001). Mean basal gastric acid output (1.6 vs 8.0 mEq/h; p < 0.001) and maximal acid output (20.6 vs 106.6 mEq/h; p < 0.001) decreased after omeprazole administration. Zinc absorption decreased after omeprazole administration (141 [34] mg/dL/h) compared with pre-omeprazole values (245 [35]; p < 0.01). CONCLUSION: Suppression of gastric acid secretion by omeprazole reduces intestinal absorption of zinc. PMID- 12546171 TI - Efficacy and safety of oral sodium phosphate versus polyethylene glycol solution for bowel preparation for colonoscopy. AB - OBJECTIVE: To assess the acceptability, safety and effectiveness of an oral sodium phosphate solution (Exelyte) for colon preparation prior to colonoscopy, compared with a polyethylene glycol solution (Peglec). METHOD: A colonoscopist blinded, prospective, randomized, observational clinical study. PATIENTS: One hundred patients undergoing colonoscopy for various indications were randomized (n = 50 each) to receive either 90 mL of oral sodium phosphate mixed with 300 mL clear liquid and then consume about 4-5 glasses of water, or 2 liters of polyethylene glycol solution. RESULT: Sodium phosphate showed a safety profile similar to that of polyethylene glycol. However, patients tolerated it better. The colonoscopist reported similar cleansing of bowel in both groups. CONCLUSION: Oral sodium phosphate is well tolerated, is safe and provides bowel cleansing similar to that with a polyethylene glycol solution. PMID- 12546172 TI - Laparoscopic versus open surgical closure of perforated duodenal ulcers: a comparative study. AB - BACKGROUND: Laparoscopic closure of duodenal ulcer perforation may be an alternative to open surgery due to lower morbidity. Most published series have used omental plug for laparoscopic closure. We performed simple closure of the perforation laparoscopically and compared the results with those obtained by open surgery. METHODS: Of 77 consecutive patients with duodenal ulcer perforation 10 were excluded due to their high risk for laparoscopic surgery. 34 (age 18-61 years; one woman) were treated by laparoscopic surgery while 33 (age 23-63 years; two women) underwent laparotomy. Closure of the perforation was achieved by suturing the edges of the perforation. RESULTS: 27 patients had successful closure of perforation by laparoscopy; one had sealed perforation and did not need closure. Conversion to open surgery was necessary in 6 patients (17.8%). Median operating time was 50 minutes (range 25 to 120) and median hospital stay was 4 days (range 4 to 6) for laparoscopy. There was no postoperative leak. Corresponding figures for open surgery were 55 minutes (45 to 75) and 9 days (7 to 13). Patients in the laparoscopy group returned early to work (median 13 days, range 10 to 15 days postoperatively) as against 26 days (21 to 35) in the open surgery group (p < 0.001). CONCLUSION: Laparoscopic closure of duodenal ulcer perforation is safe and effective. It is a better method of treating duodenal ulcer perforation when the patient's condition allows pneumoperitoneum and laparoscopy. PMID- 12546174 TI - Carcinoma of stomach in a patient with familial tylosis. AB - The association of tylosis with esophageal cancer has been extensively reported but association with gastric cancer is rare. We report a 55-year-old man with familial tylosis and carcinoma of the stomach for which radical gastrectomy was done. Repeat endoscopy 3 years later is normal. PMID- 12546173 TI - Pain in abdomen--do not forget lead poisoning. AB - Lead toxicity has been recognized for thousands of years, and is still around. We encountered 11 patients with lead toxicity in the last two years. All patients had presented with diffuse pain in the abdomen, anemia and mild derangements of liver biochemistry. History of intake of indigenous or herbal medicine for diabetes mellitus or psychosexual disorders was present in eight patients. All of them had elevated blood lead levels. Abdominal pain responded promptly to treatment with chelating agents. PMID- 12546175 TI - Nonfunctioning islet cell tumor presenting with ascites and portal hypertension. AB - Nonfunctioning islet cell tumors commonly cause no symptoms. A 22-year-old woman presented with lump in the left hypochondrium, refractory high-protein ascites and evidence of left-sided portal hypertension. At exploratory laparotomy, a 30 cm x 15 cm mass was seen at the splenic hilum, with large collateral vessels around. Distal pancreatectomy with splenectomy was done. Histology of the mass showed malignant islet cell tumor infiltrating the spleen. The patient died in the postoperative period. PMID- 12546176 TI - Tail gut cyst. AB - The tail gut is a blind extension of the hindgut into the tail fold just distal to the cloacal membrane. Remnants of this structure may form tail gut cyst. We report a 14-year-old girl with tail gut cyst that presented as acute abdomen. The patient recovered after cyst excision. PMID- 12546177 TI - Recurrent gastric trichobezoar. AB - Trichobezoars are concretions of ingested hair that are found in the stomach. Recurrence of this condition has not been reported. We report an 18-year-old girl with recurrent trichobezoar; this emphasizes the need for counseling in these patients. PMID- 12546178 TI - Intra-arterial contrast-enhanced spiral CT: adjunct to angiography for localizing obscure gastrointestinal bleed. AB - We report two patients with obscure GI bleed in whom we performed intra-arterial contrast-enhanced spiral CT scan of abdomen with the angiography catheter in the superior mesenteric artery (SMA). The technique localized the lesions to the mid jejunum and distal duodenum. PMID- 12546179 TI - Gastric mucormycosis. AB - Systemic mucormycosis is a rare fatal fungal infection that usually involves the nasopharynx. Gastrointestinal mucormycosis is rare, occurring in immunocompromised conditions and with advanced malignancies. We report a 35-year old man, an alcoholic, admitted with acute abdomen. Endoscopy revealed an ulcerated plaque-like lesion in the stomach. Histology revealed mucormycosis of the stomach. The patient successfully underwent treatment with amphotericin-B. PMID- 12546180 TI - Non-gestational choriocarcinoma in small intestine. AB - Choriocarcinoma most commonly arises from intrauterine gestational trophoblastic tissue; non gestational choriocarcinoma is rare. We report a 22-year-old married woman with non-gestational choriocarcinoma in the small intestine. Partial resection of the jejunum and ileum was done, followed by chemotherapy. She was well one year later. PMID- 12546181 TI - Acute intestinal obstruction due to small gut hemangioma. AB - A 20-year-old man presented with acute intestinal obstruction due to multiple hemangiomas of small intestine extending into the adjoining mesentery. The diagnosis was made at laparotomy and subsequently confirmed on histology. Occurrence of hemangioma in the small intestine and its presentation as acute intestinal obstruction are rare. PMID- 12546182 TI - Functional dyspepsia in Turkish children. PMID- 12546183 TI - Recurrent alveolar hydatid disease of liver. PMID- 12546184 TI - Hydatid-specific IgE and indirect hemagglutination tests may not differentiate cystadenoma and hydatid cyst. PMID- 12546185 TI - Spleen should be sacrificed for repair of traumatic rupture of stomach fundus. PMID- 12546186 TI - An accidentally swallowed toothbrush. PMID- 12546187 TI - Multisepate gall bladder: sonographic and computed tomographic appearances. PMID- 12546188 TI - Winter mortality and its causes. AB - In the 1970s scientific research focussed for the first time on dramatic rises in mortality every winter, and on smaller rises in unusually hot weather. Following the recent decline in influenza epidemics, approximately half of excess winter deaths are due to coronary thrombosis. These peak about two days after the peak of a cold spell. Approximately half the remaining winter deaths are caused by respiratory disease, and these peak about 12 days after peak cold. The rapid coronary deaths are due mainly to haemoconcentration resulting from fluid shifts during cold exposure; some later coronary deaths are secondary to respiratory disease. Heat related deaths often result from haemoconcentration resulting from loss of salt and water in sweat. With the possible exception of some tropical countries, global warming can be expected to reduce cold related deaths more than it increases the rarer heat related deaths, but statistics on populations in different climates suggest that, given time, people will adjust to global warming with little change in either mortality. Some measures may be needed to control insect borne diseases during global warming, but current indications are that cold will remain the main environmental cause of illness and death. Air pollution in cities may also still be causing some deaths, but these are hard to differentiate from the more numerous deaths due to associated cold weather, and clear identification of pollution deaths may need more extensive data than is currently available. PMID- 12546189 TI - Arctic Monitoring and Assessment Programme (AMAP); strategy and results with focus on the human health assessment under the second phase of AMAP, 1998-2003. PMID- 12546190 TI - Greenlandic family structure and communication with parents: influence on schoolchildren's drinking behaviour. AB - OBJECTIVES: The objective of this paper is to investigate how changes in the Greenlandic family structure and perceived difficulties in communicating with parents affect the prevalence of alcohol consumption and the risk of drunkenness among schoolchildren. STUDY DESIGN: The survey was carried out in February 1998 in all schools in Greenland in the context of the WHO Health Behaviour in School Aged Children (HBSC) study. Altogether 3,081 questionnaires were returned, which gave a response rate of 68% of the total study population. The present sample consists of 1,648 students, 826 boys and 822 girls in the age groups 11, 13 and 15 years. Descriptive statistics and a logistic regression model were applied. RESULTS: The results showed that the risk of drinking alcohol among Greenlandic schoolchildren increased when they were living in a broken/restructured family setting, as a single mother home or living with mother and a stepfather. This risk was higher when communication with parents was regarded by the children as being poor. The risk of drunkenness increased with age. Different age groups are influenced by different factors as regards their drinking behaviour. CONCLUSIONS: The analysis showed that alcohol drinking rose when children were living in broken families but neither gender differences of influencial factors to being drunk nor differences in drinking behaviour between social classes were found. PMID- 12546191 TI - Iodine content of traditional Greenlandic food items and tap water in East and West Greenland. AB - OBJECTIVES: The iodine intake level is important for the occurrence of thyroid disorders in a population. The iodine intake in Greenland has been proposed to be more than ten times the recommended level. However, no measurements have been performed to determine the iodine content of Greenlandic food items, drinking water, and beverages available in East and West Greenland. STUDY DESIGN: Food samples were collected at the local market, kalaalimineerniarfik, in Nuuk and Ammassalik, and tap water was obtained from all towns in Greenland. Beverages were purchased at Kalaallit Niuerfiat KNI Pisiniarfik. RESULTS: Iodine content of seal, whale, wild fowl, reindeer, and musk ox varied between 4 and 195 microg/kg with low values for terrestrial animals (< 10 microg/kg) and higher values for marine animals (10-195 microg/kg). The iodine content of fish varied from 9 microg/kg in freshwater fish to 1,380 microg/kg in a sample of cod. The iodine content of sea mammals was: blubber 130 microg/kg; viscera 70 microg/kg; meat 21 microg/kg. No difference was observed between animals from East Greenland and West Greenland (P > 0.1). Iodine content of tap water was below 3.3 microg/l for all towns. Two sorts of beer had a high iodine content, up to 240 microg/l. The iodine content of all other beverages was 5-38 microg/l. CONCLUSIONS: We found a relatively high iodine content in marine animals but low iodine content in tap water and beverages in Greenland. The food and drinking water evaluated in the present study indicate adequate iodine intake in this area and do not support the notion that Greenland is an area of excessive iodine intake. PMID- 12546192 TI - Spina bifida, folate metabolism, and dietary folate intake in a Northern Canadian aboriginal population. AB - OBJECTIVES: Inhabitants of the subarctic region of the Eastern James Bay of Northern Quebec consume a diet low in folate. This is largely secondary to poor access to plant-foods and a preferred diet high in meat, fowl, and fish as in many other northern populations. Furthermore, there is a high frequency of spina bifida in the Cree of the region. It was hypothesized that genetically altered folate metabolism as well as low folate intake contributes to the high frequency of spina bifida. METHODS: A case-control study evaluating folate metabolism and the common 677C-T polymorphism of the gene for methylenetetrahydrofolate reductase (MTHFR) in mothers of children with spina bifida, and controls (n=23) of Cree descent from the Eastern James Bay region. These results were compared to a similar Montreal cohort (n=152) who were not of First Nations descent. Dietary intake of folate of 219 women of the Eastern James Bay region was also determined. RESULTS: No Cree mothers of children with spina bifida were homozygous for the 677C-T polymorphism of MTHFR. Although serum cobalamin was significantly higher in Cree mothers, RBC folate was significantly lower than in the Montreal cohort. In addition, plasma homocysteine was significantly lower in the Cree. Dietary intake of folate of women in the same region was substantially lower (100 microg/day) than widely recommended daily intakes. CONCLUSIONS: In this remote Canadian aboriginal community there is no evidence of altered folate metabolism in the mothers of children with spina bifida. Nonetheless, it remains essential that culturally appropriate public health efforts be continued to increase the intake of folic acid in the hope of reducing the high frequency of spina bifida in this population. PMID- 12546193 TI - Incidence of frostbite and ambient temperature in Finland, 1986-1995. A national study based on hospital admissions. AB - OBJECTIVES: The association of frostbite with ambient temperature in Finland is not known. The present study determined the incidence of frostbite and its association with sex, age and ambient temperature in a nationwide sample. STUDY DESIGN: The first admissions of all patients (n=1,275) admitted to hospital in Finland during the period 1986-1995 with frostbite as a principal or secondary diagnosis were associated with ambient temperature on the day of admission. RESULTS: The incidence of frostbite was 2.5 per 100,000 inhabitants, it was higher in males than females and increased linearly with age. The annual incidence of frostbite started to rise at below -15 degrees C and was considerable at under -20 degrees C, particularly in northern Finland. However, the daily incidence increased most in the urban area of Helsinki. CONCLUSION: In the north, the main factor is the large number of cold days in the year. The daily incidence may be affected by urban lifestyle, possibly fashion, and inability to protect oneself against the cold. PMID- 12546194 TI - Plasma catecholamines, serotonin and their metabolites and beta-endorphin of winter swimmers during one winter. Possible correlations to psychological traits. AB - OBJECTIVES: The study was a follow-up one, in which blood pressure and hormonal changes were investigated during one winter swimming season in winter swimmers (WSs) and non-swimmer controls on three occasions (autumn, winter and spring). Humoral results were compared to psychological traits recorded at the time of the three blood samplings. RESULTS: Mean systolic blood pressure of the WSs fell from 134 +/- 12 mmHg to 128 +/- 12 mmHg (p < 0.05) during the winter, and a slight but non-significant drop was also seen in the controls. Mean plasma noradrenaline concentrations diminished significantly from autumn to spring, and more so in the WS-group, but no statistically significant difference was observed between the groups. Adrenaline levels also showed a decreasing trend, and the change was significant when calculated by using the combined means of both groups. Plasma homovanillic acid and beta-endorphin values were on the same level in all seasonal samples in both groups. Plasma serotonin levels decreased in both groups by about 50 per cent by spring, but 5-HIAA did not change significantly. HVA showed correlation with blood pressure and anxiety in the autumn (r=0.367). In the winter measurement endorphin and hysteria had a negative correlation (r=0.370). In the spring 5-HIAA and obsessionality had a positive correlation (r=0.351). DISCUSSIONS: In summary, blood pressure and plasma catecholamine levels decreased during winter swimming practice over one winter, but these changes were also observed in the control persons. Plasma serotonin was lower in the spring in both groups. The changes in the humoral status speak for adaptation to the research situation, or reflect seasonal variation from autumn to spring. No clear effect of winter swimming as such was detected. PMID- 12546195 TI - Cold and the risk of cardiovascular diseases. A review. AB - The higher occurrence of cardiovascular diseases in winter is well known, and several explanatory mechanisms have been suggested based on increased blood pressure, haematological changes and respiratory infections. Most investigations have used ecological data such as daily temperatures recorded at weather stations and mortality in the general population. Cause-specific mortality is the outcome measure most commonly used. Local myocardial infarction community registers would offer an ideal database, but may suffer from inadequate statistical power. Hospital discharge records, linked with out-of-hospital deaths, provide a powerful tool for detecting even weak effects of temperature. The association of coronary heart disease and temperature is usually U-shaped, mortality being lowest within the range 15-20 degrees C and higher on both sides of this. The increase in mortality on the colder side is in the region of 1% per 1 degree C fall in temperature, but the increase on the warmer side may be very steep. The exact location of the minimum temperature and the magnitude of the effect can vary between countries. In Finland the winter excess mortality from coronary heart disease has been levelling off during recent decades, but it still represents approximately 6% of annual deaths due to this condition. PMID- 12546196 TI - The lactic acid bacteria: a literature survey. AB - The purpose of this review article on the lactic acid bacteria grew from an early curiosity and a desire to convey and impart the broad scope of literary information on their functions as starter cultures, in the manufacture of fermentation products such as dairy products and alcoholic beverages, as well as their contribution to better health. This review article is an attempt to empower the reader and to circumvent the difficult task in acquiring and elucidating a large body of information. The intent is to familiarize the reader with the various lactic species, their habitat or source, associated food, physiological characteristics, colonial morphology, biochemical characteristics, culture media (enrichment, nonselective, and selective), classic description, and taxonomy. This review provides information on Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, Carnobacterium, and Enterococcus. Trends are presented, such as the use of nisin to extend food shelf-life and the current research premise that Probiotic strains may alter the intestinal flora and thus prevent intestinal wall penetration by pathogens. PMID- 12546197 TI - Microbial agents associated with waterborne diseases. AB - Many classes of pathogens excreted in feces are able to initiate waterborne infections. There are bacterial pathogens, including enteric and aquatic bacteria, enteric viruses, and enteric protozoa, which are strongly resistant in the water environment and to most disinfectants. The infection dose of viral and protozoan agents is lower than bacteria, in the range of one to ten infectious units or oocysts. Waterborne outbreaks of bacterial origin (particularly typhoid fever) in the developing countries have declined dramatically from 1900s. Therefore, some early bacterial agents such as Shigella sonnei remains prevalent and new pathogens of fecal origin such as zoonotic C. jejuni and E. coli O157:H7 may contaminate pristine waters through wildlife or domestic animal feces. The common feature of these bacteria is the low inoculum (a few hundred cells) that may trigger disease. The emergence in early 1992 of serotype O139 of V. cholerae with epidemic potential in Southeast Asia suggests that other serotypes than V. cholerae O1 could also getting on epidemic. Some new pathogens include environmental bacteria that are capable of surviving and proliferating in water distribution systems. Other than specific hosts at risk, the general population is refractory to infection with ingested P. aeruginosa. The significance of Aeromonas spp. in drinking water to the occurrence of acute gastroenteritis remains a debatable point and has to be evaluated in further epidemiological studies. Legionella and Mycobacterium avium complex (MAC) are environmental pathogens that have found an ecologic niche in drinking and hot water supplies. Numerous studies have reported Legionnaires' disease caused by L. pneumophila occurring in residential and hospital water supplies. M. avium complex frequently causes disseminated infections in AIDS patients and drinking water has been suggested as a source of infection; in some cases the relationship has been proven. More and more numerous reports show that Helicobacter pylori DNA can be amplified from feces samples of infected patients, which strongly suggests fecal to-oral transmission. Therefore, it is possible that H. pylori infection is waterbome, but these assumptions need to be substantiated. Giardiasis has become the most common cause of human waterborne disease in the U.S. over the last 30 years. However, as a result of the massive outbreak of waterborne cryptosporidiosis in Milwaukee, Wisconsin, affecting an estimated 403,000 persons, there is increasing interest in the epidemiology and prevention of new infection disease caused by Cryptosporidium spp. as well as monitoring water quality. The transmission of Cryptosporidium and Giardia through treated water supplies that meet water quality standards demonstrates that water treatment technologies have become inadequate, and that a negative coliform no longer guarantees that water is free from all pathogens, especially from protozoan agents. Substantial concern persists that low levels of pathogen occurrence may be responsible for the endemic transmission of enteric disease. In addition to Giardia and Cryptosporidium, some species of genera Cyclospora, Isospora, and of family Microsporidia are emerging as opportunistic pathogens and may have waterborne routes of transmission. More than 15 different groups of viruses, encompassing more than 140 distinct types can be found in the human gut. Some cause illness unrelated with the gut epithelium, such as Hepatitis A virus (HAV) and Hepatitis E virus (HEV). Numerous large outbreaks have been documented in the U.S. between 1950 and 1970, and the incidence rate has strongly declined in developing countries since the 1970s. Hepatitis E is mostly confined to tropical and subtropical areas, but recent reports indicate that it can occur at a low level in Europe. A relatively small group of viruses have been incriminated as causes of acute gastroenteritis in humans and fewer have proven to be true etiologic agents, including rotavirus, calicivirus, astrovirus, and some enteric adenovirus. These enteric viruses have infrequently been identified as the etiologic agents of waterborne disease outbreaks, because of inadequate diagnostic technology, but many outbreaks of unknown etiology currently reported are likely due to viral agents. Actually, Norwalk virus and Norwalk-like viruses are recognized as the major causes of waterborne illnesses world-wide. The global burden of infectious waterborne disease is considerable. Reported numbers highly underestimate the real incidence of waterborne diseases. The most striking concern is that enteric viruses such as caliciviruses and some protozoan agents, such as Cryptosporidium, are the best candidates to reach the highest levels of endemic transmission, because they are ubiquitous in water intended for drinking, being highly resistant to relevant environmental factors, including chemical disinfecting procedures. Other concluding concerns are the enhanced risks for the classic group of debilitated subjects (very young, old, pregnant, and immunocompromised individuals) and the basic requirement of to take specific measures aimed at reducing the risk of waterborne infection diseases in this growing, weaker population. PMID- 12546198 TI - Putting public health practice into primary care practice: practical implications of implementing the changes in shifting the balance of power in England. PMID- 12546199 TI - A CLAS act? Community-based organizations, health service decentralization and primary care development in Peru. Local Committees for Health Administration. AB - In 1994 Peru embarked on a programme of health service reform, which combined primary care development and community participation through Local Committees for Health Administration (CLAS). They are responsible for carrying out local health needs assessments and identifying unmet health needs through regular household surveys. These enable them to determine local health provision and tailor services to local requirements. CLAS build on grassroots self-help circles that developed during the economic and political crises of the 1980s, and in which women have been prominent. However, they function under a 3 year contract with the Ministry of Health and within a framework of centrally determined guidelines and regulations. These reforms were implemented in the context of neo-liberal economic policies, which stressed financial deregulation and fiscal and monetary restraint, and were aimed at reducing foreign indebtedness and inflation. We evaluate the achievements of the CLAS and analyse the relationship between health and economic policy in Peru, with the aid of two contrasting models of the role of the state - 'agency' and 'stewardship'. We argue that Peru's experience holds valuable lessons for other countries seeking to foster community involvement. These include the need for community capacity building and partnership between community organizations and state (and other civil) agencies. PMID- 12546200 TI - Availability of primary care doctors and population health in England: is there an association? AB - BACKGROUND: In the United States, an association has been proposed between better access to primary care and lower mortality. This paper reports an ecological analysis that evaluated whether population health was associated with general practitioner (GP) supply in England. METHODS: Data were analysed for 99 health authorities in England in 1999. Health outcomes included standardized mortality ratios, infant mortality rate (per 1,000), hospital admissions with acute and chronic conditions (per 100,000), and teenage conception rates (per 1,000). The number of GPs per 10,000 population was included as explanatory variable. Confounders included the Townsend deprivation score, proportion of ethnic minorities, proportion in social classes IV and V, and proportion with limiting long-term illness. Analyses were by linear regression weighted for population size. RESULTS: Higher GP supply was associated with lower mortality in univariate analyses. After adjusting for deprivation score, ethnic group and social class, the standardized mortality ratio for all-cause mortality at 15-64 years decreased by -5.2 (95 per cent confidence interval -8.3 to -2.0, p = 0.002) per unit increase in GP supply. After additional adjustment for limiting long-term illness, the decrease was -3.3 (-6.7 to 0.1, p = 0.060). In the fully adjusted model, each unit increase in GP supply was associated with a decrease in hospital admission rates for acute conditions (-14.4, -21.4 to -7.4 per 100,000, p < 0.001) and chronic conditions (-10.6, -17.2 to -4.0, p = 0.002). CONCLUSIONS: In England, lower supply of GPs was associated with increased hospital utilization, but a strong univariate association with mortality might be explained by confounding. PMID- 12546201 TI - Leukaemia incidence near coastal features. AB - BACKGROUND: The aim of the study was to independently test the hypothesis that leukaemia incidence is higher in proximity to estuaries. METHODS: Electoral wards were classified as to whether they included estuarine, coastal or only inland features. Rates of different adult and childhood leukaemias were computed for each ward category; that is, acute lymphoblastic leukaemia (ALL), acute myeloid leukaemia (AML), chronic myeloid leukaemia (CML) aged 0-79 and for all childhood leukaemias combined (aged 0-14). RESULTS: Poisson regression analysis controlling for the effects of sex, age, and socioeconomic and urban-rural status, showed no statistically significant differences in incidence between wards with different levels of estuarine classification. CONCLUSION: The hypothesis created from an earlier dataset that a link exists between leukaemia and residence near estuaries is not upheld. PMID- 12546202 TI - Evaluating the quality of internet-based information about alternative therapies: development of the BIOME guidelines. AB - BACKGROUND: The aim of the study was to develop guidelines for evaluating the quality of Internet-based information about alternative therapies. METHOD: An expert committee drafted a set of guidelines for evaluating information relating to alternative therapies. The guidelines were subsequently refined by testing them using resources already included in the BIOME databases. The first 20 unique web sites about alternative therapies for cancer retrieved using a general search engine and a United Kingdom focused search engine were then evaluated using the refined guidelines. Those undertaking the evaluations also completed a questionnaire relating to the face and content validity of the guidelines. The participants in the implementation stage were six content providers. Content providers identify, evaluate and describe resources for inclusion in the BIOME databases. RESULTS: Only one web site out of 20 was selected by all six content providers for inclusion in the BIOME databases according to the alternative therapies guidelines. All content providers were in agreement regarding the exclusion of nine sites, but there were discrepancies regarding the remaining 10 resources. There was general agreement that the guidelines were easy to understand and that all points raised were necessary. However, there were differences of opinion regarding whether all issues were covered, whether the guidelines allowed the selection of only the highest quality resources, and whether the guidelines were applicable to a wide range of Internet-based resources about alternative therapies. CONCLUSIONS: The levels of inconsistencies in the results indicate the need for the further development of the BIOME guidelines for selecting information about alternative therapies. PMID- 12546203 TI - Trends in the incidence of cutaneous malignant melanomas in the south east of England, 1960-1998. AB - BACKGROUND: Our aim was to describe the epidemiology of cutaneous malignant melanomas (CMMs) in males and females in the south east of England from 1960 to 1998. Birth cohort effects are important when describing the incidence of CMMs because fashions of dress affect whether different body sites are exposed to sunlight. METHODS: We calculated the age-standardized incidence of all CMMs for 5 year periods from 1960 to 1998 and the age-specific and age-standardized incidence of CMMs at different sites for 5 year periods from 1980 to 1998. We plotted age-specific incidence rates against period of diagnosis and birth cohort. RESULTS: The age-standardized incidence of CMMs increased from 1.3 to 7.1 per 100,000 person-years in males. Incidence increased from 2.4 to 7.8 per 100,000 person-years overall in females but levelled in the 1990s. Trunk CMMs had the highest incidence in males and the greatest increase in incidence. Lower limb CMMs had the highest incidence in females but their incidence decreased. The rate of increase of the age-specific incidence of trunk CMMs was greater for male birth cohorts before about 1945. The rate of increase of the incidence of lower limb CMMs was greater for female birth cohorts before about 1920. CONCLUSIONS: If current trends continue, the age-standardized incidence of CMMs in females will remain stable. The increase in the age-standardized incidence of CMMs in males was driven by increases in the age-specific incidence of trunk CMMs in men who were born before about 1949. As they age, age-standardized incidence should level. PMID- 12546204 TI - Social and demographic predictors of parental consultation for child psychological difficulties. AB - BACKGROUND: Results from previous studies examining determinants of parental consultation for child mental health provide inconsistent evidence concerning socio-demographic predictors. The aim of this study is to identify the sociodemographic predictors of parental consultation for child psychological difficulties. METHOD: An epidemiological cross-sectional analysis was carried out using a sample of 5,913 children aged between 4 and 15 years from the Health Survey for England. The Strengths and Difficulties Questionnaire (SDQ) was the measure of child psychological morbidity. RESULTS: Parents of children with psychological difficulties were less likely to seek a consultation if their child was a girl, as household income decreased or if the head of household came from manual social class. In contrast, parents were more likely to seek a consultation if they were in receipt of a benefit than if they were not in receipt of a benefit. Age of child and family type did not predict parental consultation. CONCLUSIONS: The results of this analysis confirm that a substantial proportion of children with mental health difficulties in the general population (42 per cent) have not been seen by a professional, and these are likely to be girls and children in low-income families, indicating a significant unmet need for services across the nation. These results suggest that parents and health professionals should be made more aware of the symptoms of psychological problems in girls and that services need to be planned in a way that improves uptake by low-income parents. PMID- 12546205 TI - Non-attendance at re-examination 20 years after screening in the British Regional Heart Study. AB - BACKGROUND: Maintenance of high participation rates in longitudinal studies is critical to their validity because of the possibility of bias associated with non participation, which may differ between studies. This paper examines factors associated with participation status over time, by comparing the characteristics of non-attenders and attenders at a 20 year follow-up examination in a cardiovascular cohort study with an initial participation rate of 78 percent. METHODS: A baseline examination was carried out between 1978 and 1980 and subjects have been followed up through regular reviews of general practice records, postal questionnaires, and a clinical re-examination of survivors in 1998-2000. Data obtained by questionnaire in 1996 (Q96) and at baseline examination have been used to compare the characteristics of these recent non attenders and attenders who remain in the cohort for further follow-up. RESULTS: The non-attendance rate of available survivors at re-examination was 23 per cent (n = 1313). Rates of non-attendance were highest in Scotland and lowest in Southern England. Non-attenders were older than attenders, more likely to smoke, and reported more disabling conditions and greater use of multiple medications at Q96, but recalled similar rates of diagnosed coronary heart disease. Marked differences in social factors were identified. Total mortality rates within 1 year of re-examination were over three times higher amongst non-attenders than attenders. CONCLUSIONS: Non-attendance is related to health status, risk factor status and social circumstances and may affect some estimates of disease prevalence, but does not appearto have a major impact on the estimated prevalence of coronary heart disease. Information collected on non-attenders in longitudinal studies helps to estimate, adjust and minimize these effects. PMID- 12546206 TI - Information for clinical governance: analysis of routine hospital activity data in Wales. AB - BACKGROUND: Variations in hospital admission rates have been extensively reported for many years, but this evidence has not had a wide impact on clinical practice. Understanding local reasons for high variation to improve quality of healthcare should be a focus of clinical governance. Our aim was to convert routine hospital activity data into information on a category of high-variation, discretionary, hospital admissions and provide a tool for analysis for clinical governors in Local Health Groups (LHG). METHODS: We undertook a cross-sectional analysis of hospital activity data for the 22 LHGs in Wales and 101 general practices in Gwent Health Authority. Hospital spells for 1998-1999 and 1999-2000 were classified into Healthcare Resource Groups (HRGs). Using the systematic component of variation we identified a category of high-variation admissions for which the only plausible explanation was medical discretion. Using scatter plots we compared the proportion of these discretionary admissions with the age-, sex- and deprivation-adjusted standardized admission ratio. (SAR) for each LHG and practice. RESULTS: We found a two-fold variation in SARs between LHGs and a three fold variation between practices. Mean discretionary activity was 55 per cent (range 50-59 per cent) of total activity for LHGs and 56 per cent (51-62 per cent) for practices. Greatest variation was found for elective admissions. The relation between discretionary admissions and the SAR was identified for each LHG and practice as the starting point for further investigation. CONCLUSION: This method provides useful information to LHG clinical governors to contribute to the process of reducing medical practice variation, increasing equity, improving the quality of care and making more cost-effective use of resources. PMID- 12546207 TI - The impact of a television soap opera on the NHS Cervical Screening Programme in the North West of England. AB - BACKGROUND: Mass media interventions can influence health care utilization but the effect of televised fictional accounts of illness upon national screening programmes is unknown. Our aim was to evaluate the impact of a Coronation Street story line, in which one of the characters died from cervical cancer, on the National Health Service (NHS) Cervical Screening Programme. METHODS: The study involved a retrospective analysis of information held on cervical screening databases ('Exeter' computer systems) of the nine Health Authorities constituting the Lancashire and Greater Manchester zones of the North West Region of the NHS. The number of cervical smears performed in the community, in women over 25 years of age, whose previous smear was normal and who were on routine recall, during a 6 month period that included the story line, was compared with those taken over the same period in the previous year. The proportions of smears classified by a screening interval of 'unscheduled', 'on time', 'overdue' or 'no previous smear' were compared. RESULTS: The number of smears performed increased from 65,714 in 2000 to 79,712 in 2001, an increase of 13,998 (21.3 percent; 95 per cent confidence interval (CI) 21.0-21.6 per cent) in the 19 weeks after the story line. The increase in the number of smears occurred in all categories of screening interval, with the largest increase seen in those attending 'on-time' (26 per cent). CONCLUSIONS: We have demonstrated a large impact of a soap opera story line on the cervical screening programme although the benefit to health is not clear. Further research will determine the long-term effect of the story. PMID- 12546208 TI - The effect of 'Alma's' death on women attending for a cervical smear: a questionnaire survey. AB - Following a story line in Coronation Street in which 'Alma' died of cervical cancer there was a large increase in the number of cervical smears taken in Manchester. To ascertain the extent to which women were influenced to attend for smears by this story line and why, we carried out a postal survey. Our results show that women were influenced to attend for smears (22 per cent) with many women doing so because the story line made them worry. Television influenced screening behaviour but it must be of concern to health promoters that this was done through the generation of anxiety. PMID- 12546209 TI - Prescribing welfare benefits advice in primary care: is it a health intervention, and if so, what sort? AB - There is increasing provision of welfare benefits advice in primary care (WBAPC). This reflects the present government's recognition of the association between socioeconomic and health inequalities. However, the assumption that increasing an individual's income will improve their health is not based on clear evidence. This paper reviews the relevant evidence, using four categories of pathways from poverty to ill-health: individual material; environmental material; individual psychological; environmental psychological. A description is offered of the scope and limits of WBAPC as a health intervention: one that primarily offers relief from psychological stress for service users, who tend to be middle-aged or older and suffering from chronic disease or disability. WBAPC may also make a small contribution to the physical health of individuals and to the social capital of communities. Thus to define the scope of WBAPC does not diminish its value to its beneficiaries, who carry a significant burden of both poverty and illness, nor does it in any way weaken the ethical argument for public policy to seek to reduce inequalities, and for primary care to play its part in that. PMID- 12546210 TI - South Asian ethnicity and risk of childhood accidents: an ecological study at enumeration district level in Leicester. AB - BACKGROUND: Surveys of 'self-reported' accidents suggest that South Asian children in the United Kingdom may have lower rates of childhood accidents, but little is known about their susceptibility to severe accidents compared with white children. METHODS: We conducted an ecological study at the level of Census enumeration districts to compare hospital utilization as a result of childhood accidents according to White, South Asian, Black or 'Other' ethnic grouping and Townsend deprivation score in Leicester. Enumeration districts were assigned to postcoded data for fracture clinic attendances between 1997 and 1999 and in patient admissions and in-patient stays of longer than 3 days as a result of accidents between 1995 and 1999 in children under 16 years. RESULTS: South Asian children were less likely than white children to attend fracture clinic, be admitted or to have a prolonged stay as a result of an accident. Having adjusted for deprivation score, for a 10 per cent increase in the proportion of South Asian residents in an enumeration district, the odds ratio for an in-patient stay of longer than 3 days was 0.95 (95 per cent confidence interval (CI) 0.91-1.00, p = 0.035), for an accident admission the odds ratio was 0.93 (95 per cent CI 0.92 0.94, p < 0.001) and for attendance at fracture clinic the odds ratio was 0.94 (95 per cent CI 0.92-0.96, p < 0.001). For a district with 70 per cent of its children from South Asian groups (as observed in one-fifth of Leicester's enumeration districts), this represents a 40 per cent lower rate of accident admissions. CONCLUSIONS: South Asian children were significantly less likely to utilize hospital services as a result of an accident. This may well be explained by differential exposure to accident hazards across ethnic groups, rather than by different thresholds of hospital attendance, given that hospital utilization was also lower for serious accidents in South Asian children. PMID- 12546211 TI - Views of Directors of Public Health about NICE Appraisal Guidance: results of a postal survey. National Institute for Clinical Excellence. AB - BACKGROUND: We aimed to determine the views of Directors of Public Health about the Health Technology Appraisal Programme of the National Institute for Clinical Excellence (NICE) before the move to strategic health authorities and primary care trusts in April 2002. METHOD: In December 2001 we sent a questionnaire asking about the work programme, products, decision-making, general approach, resource allocation and success of NICE to all Directors of Public Health in England and Wales. Ninety-two of 100 responded. RESULTS: Three-quarters or more agreed that NICE has covered a number of priority and controversial areas, produced good-quality health technology appraisals, well-presented reports and readable guidance in a consistent format, that it has raised the profile of clinical effectiveness, provided a focus for debate about health technology, and succeeded in making the National Health Service (NHS) set aside resources for approved technologies. A similar proportion, however, also agreed that guidance was not timely, did not address 'whole systems' and made some disappointing recommendations, and that decision-making was not influenced enough by the needs of the NHS. They considered that NICE did not address implementation, decide between competing technologies or help the service prioritization debate, and that guidance sent unrealistic signals about affordability to patients and politicians and caused difficulty for the implementation of other technologies locally. CONCLUSIONS: A majority of Directors are positive about NICE's role of providing high-quality appraisal and central guidance but negative about its influence on local priority setting. Major concerns remain about the affordability of competing demands, whether this is NICE's responsibility or not. PMID- 12546212 TI - The ability of general practitioners to detect mental disorders among primary care patients in a stressful environment: Gaza Strip. AB - BACKGROUND: The aim of the present study was to investigate the detection rate by general practitioners (GPs) of mental disorders in a primary health care setting and relating the findings to selected GP characteristics and the patient sociodemographic characteristics. METHODS: The patients were assessed with respect to mental disorders by Hopkins Symptom Checklist 25 (HSCL-25), and the GPs were independently asked to fill in the Goldberg checklist II to assess the patient after consultation. The sample consisted of 10 primary health care clinics in the Gaza Strip, which were randomly selected from the five regions that form the Gaza Strip (Northern, Southern region, Gaza City, Middle region, Khan-Younis and Rafah). Thirty-two GPs and 661 patients participated in the study. RESULTS: The study showed that the GPs detected only 11.6 per cent of patients with mental disorders at HSCL-25 score >1.75, and that the GP's assessment was not significantly associated with the HSCL-25 scores. GPs with postgraduate psychiatric training performed better in detecting mental disorders, likewise female GPs and those who were more than 40 years old. The results also revealed that the GPs were more able to detect mental disorders among patients older than 25 years, and in female patients. CONCLUSIONS: The GPs' poor detection rate of mental disorders indicates the importance of mental health training for GPs working in primary health care clinics. PMID- 12546213 TI - Alcohol awareness and unit labelling. PMID- 12546214 TI - Quarterly Communicable Disease Review April to June 2002. PHLS Communicable Disease Surveillance Centre. PMID- 12546215 TI - Future role of public health medicine. PMID- 12546216 TI - First, first among equals, or equals? Challenging the medical role in public health. PMID- 12546217 TI - The 'Seascale cluster'. PMID- 12546218 TI - Childhood cancer in Seascale. PMID- 12546219 TI - More than one reason to immunize health care workers. PMID- 12546220 TI - My favourite software: using templates in Word. PMID- 12546221 TI - Evolution in zigzag--the changing state of A-bomb dosimetry. PMID- 12546222 TI - Promoting transparency in regulation--determining radiological doses to members of critical groups. PMID- 12546223 TI - An update on the discrepancy between calculated and measured neutron-induced radioactivity levels in Hiroshima. AB - The thermal neutron activation measurements carried out over many years in Hiroshima and Nagasaki have been the subject of ongoing debate in recent years because they indicate that current DS86 neutron doses may have been significantly underestimated in Hiroshima. Long-lived neutron activation products, 60Co, 152Eu, 154Eu and 36Cl, which are still detectable today using modern analytical techniques, appear to indicate that DS86 calculated thermal neutron activation products decrease with distance more rapidly than the measured values. The latest thermal neutron activation measurements have been collated and a new relationship for the measured to calculated (M/C) ratio of induced activity has been derived as a function of slant range. This indicates a stronger dependence of M/C on slant range than previously derived by Straume et al (1992 Health Phys. 63 421-6) and emphasises even more the discrepancy between measured and calculated (DS86) neutron doses at distances beyond 1 km. While the main body of thermal neutron activation data appears to support a significant increase in the DS86 neutron dose component in Hiroshima, there are some thermal neutron activation measurements and some very recent fast neutron activation measurements which suggest that the discrepancy may not be so great. The extent of the required revision to the neutron component of the DS86 dosimetry remains the subject of ongoing new neutron activation measurements and re-analysis of existing published measurements. A companion paper considers the impact on radiation risk estimates of possible modifications to the DS86 dosimetry system on the basis of a broad range of interpretations of the neutron activation data. PMID- 12546224 TI - The impact of possible modifications to the DS86 dosimetry on neutron risk and relative biological effectiveness. AB - The current DS86 dosimetry system for the Japanese bomb survivors indicates that neutron doses were so low that they prevent the direct derivation of any useful estimates of neutron risk. However, the large body of thermal neutron activation measurements carried out over many years in Hiroshima and Nagasaki appear to indicate that current DS86 neutron doses may have been significantly underestimated in Hiroshima. An earlier companion paper has provided an update of neutron activation measurements. While a large body of data appears to support a significant increase, there is ongoing debate and review regarding its validity. However, as yet, there are no detailed, peer-reviewed, published refutations of the neutron activation data which appear to support an increase in neutron doses. In this paper, we consider the impact of possible future revisions in the DS86 dosimetry on radiation risk estimates. We consider the extreme range of possibilities from maintaining the existing DS86 values, to changes in neutron doses in accord with the majority of existing neutron activation data. We have used the latest cancer incidence data and cancer mortality data for the A-bomb survivors, and neutron doses have been modified using a neutron revision factor (NRF) in line with the latest thermal neutron activation measurements in Hiroshima. In contrast to previous analyses, a nonlinear relationship between log(NRF) and slant range has been used which better represents the data beyond slant ranges of approximately 1 km. The impact on the evaluation of neutron relative biological effectiveness (RBE) and gamma radiation risk estimates has been assessed. While DS86 neutron doses are too low to allow any useful direct evaluation of neutron risk or neutron RBE, it becomes possible to derive more meaningful values if neutron doses are increased in Hiroshima in line with the broad range of thermal neutron activation measurements. The uncertainties are smallest for the cancer incidence data. The best estimates of neutron RBE give upper 95% confidence limits of about 6 for all solid tumours for the incidence data and about 28 for the mortality data. The uncertainties in neutron RBE for leukaemia incidence are larger, and estimation at doses below about 0.1 Gy is not possible. There is no significant change in the excess relative risk for gamma radiation for all solid tumours taken together, compared with the current DS86 dosimetry. The results preclude neutron RBE values significantly greater than the current ICRP radiation weighting factors, which range between 5 and 20, depending on energy. Whether or not the Japanese bomb survivors can indeed form the basis for useful, directly determined neutron risks clearly depends on the veracity of existing neutron activation data. This is currently the subject of careful international scrutiny and the outcome is eagerly awaited. PMID- 12546225 TI - Radioactivity in food and the environment: calculations of UK radiation doses using integrated assessment methods. AB - A new method for estimating radiation doses to UK critical groups is proposed for discussion. Amongst others, the Food Standards Agency (FSA) and the Scottish Environment Protection Agency (SEPA) undertake surveillance of UK food and the environment as a check on the effect of discharges of radioactive wastes. Discharges in gaseous and liquid form are made under authorisation by the Environment Agency and SEPA under powers in the Radioactive Substance Act. Results of surveillance by the FSA and SEPA are published in the Radioactivity in Food and the Environment (RIFE) report series. In these reports, doses to critical groups are normally estimated separately for gaseous and liquid discharge pathways. Simple summation of these doses would tend to overestimate doses actually received. Three different methods of combining the effects of both types of discharge in an integrated assessment are considered and ranked according to their ease of application, transparency, scientific rigour and presentational issues. A single integrated assessment method is then chosen for further study. Doses are calculated for surveillance data for the calendar year 2000 and compared with those from the existing RIFE method. PMID- 12546226 TI - Doses to organs and tissues from radon and its decay products. AB - This paper discusses the doses from radon and from its short-lived decay products to a number of organs and tissues and to the foetus. The aim is to put all these doses into context rather than concentrating only on the largest contributions. There is also a brief discussion of the evidence from epidemiology on the risks of exposure to radon and its decay products. As is well known, under normal circumstances the greatest hazard is to the respiratory tract from inhalation of radon decay products. Radon decay products may also give substantial doses to skin. Under some circumstances it seems likely that ingested radon could give significant doses to the stomach. Other risks appear to be smaller; the results given here allow them to be compared. PMID- 12546227 TI - Radiological assessment of the level of safety in logging operations in the Nigerian petroleum industry. AB - Petroleum prospecting and producing activities have been going on in the Niger Delta area of Nigeria for about 40 years. During this period controlled substances such as chemicals and radioactive materials have been widely used in petroleum exploration and exploitation. Deviations from acceptable levels of certain parameters relevant to safety and environmental protection have been encountered, but most have not been investigated or documented. In particular, cases involving the unsafe use, loss and abandonment of radioactive materials have neither received the desired attention nor been reported. This work reports a radiological assessment of safety in the use of radioactive and radiation producing materials in logging and well study operations in the Nigerian petroleum industry. The assessment protocol used for the evaluation is based on a numerical ranking system. Based on a scale of 100, it is found in this logging and well study that the level of safety as defined in the text is around 60% for all three sites assessed. There is substantial work needed to raise the radiation protection standards further at these sites. PMID- 12546228 TI - Determination of percentage depth dose for 6 and 10 MV x-rays using Ge-doped optical fibre and TLD-100. AB - Percentage depth doses for 6 and 10 MV x-ray beams from a linear accelerator were measured using approximately 1 cm long (approximately 0.3 mg) Ge-doped optical fibre as a thermoluminescence dosimeter for two field sizes, 5 x 5 and 10 x 10 cm2. The results indicate that the Ge-doped optical fibre dosimeter is in good agreement with the results from a PTW 30001 cylindrical ionisation chamber and TLD-100. For 6 MV x-ray beams we observe that the depth of maximum dose d(max) is 1.5 and 2 cm for field sizes of 5 x 5 and 10 x 10 cm2 respectively. For 10 MV d(max) is 2 cm for a field size of 5 x 5 cm2 and 2.5 cm for a 10 x 10 cm2 field. PMID- 12546229 TI - Comments on the recent ICRP proposals. PMID- 12546230 TI - Irish Government objects to UK nuclear activities. PMID- 12546231 TI - SRP annual general meeting: The changing role of the radiation protection professional. Cardiff, 16-7 April 2002. PMID- 12546232 TI - SRP meeting: Radio wave exposures-a cause for concern? Preston, June 2002. PMID- 12546233 TI - SRP meeting: Internal dosimetry. The British Library, 23 October 2002. PMID- 12546234 TI - Asian Regional Conference on Evolution of the System of Radiological Protection. Tokyo, 24-25 October 2002. PMID- 12546235 TI - Third International Symposium on the Protection of the Environment from Ionising Radiation. Darwin, Australia, 22-26 July 2002. PMID- 12546237 TI - Sleep disorders in a military population. AB - INTRODUCTION: Sleep disorders are common in the civilian population, but little is known about which sleep disorders are common in members of the military. This article compares a group of military personnel referred to our sleep disorders center with a group of civilian controls also referred to our sleep disorders center. METHODS: We analyzed the data of 70 Canadian military personnel and 70 civilian controls matched for age and gender. All subjects had full polysomnography. We compared reasons for referral and final sleep diagnoses for both groups. RESULTS: The mean age of each group was 40.8 +/- 7.0 SD (military) and 40.8 +/- 7.3 SD (civilians), and there were 61 men and 9 women in each group. Both groups were obese (body mass index, 30.2 +/- 5.3 (military) versus 32.5 +/- 6.9 (civilian)). Both groups were also pathologically sleepy during the day (Epworth Sleepiness Score, 10.4 +/- 4.6 (military) versus 11.3 +/- 5.4 (civilian)). The majority of referrals in each group were to rule out a sleep breathing disorder (SBD) (66% military versus 79% civilian, p = not significant). Only military patients were referred to rule out a movement disorder (17.1% military versus 0% civilian; 95% confidence interval of the difference = 8.4% 27.6%, p < 0.05). Fewer military were referred because of excessive daytime sleepiness or insomnia (7.1% military versus 20.0% civilian, 95% confidence interval of the difference = -24.4% to -1.4%, p < 0.05). The most common diagnosis confirmed in both groups was a SBD (53% military, 66% civilian, p = not significant). CONCLUSIONS: The range and distribution of sleep disorders seen in the military population is similar to that in the civilian population. Both groups were overweight and sleepy and were found to have SBD and movement disorders. These findings underscore the importance of diagnosing and treating sleep disorders in both groups. The neurocognitive impairment associated with SBD and movement disorders impacts highly on the ability of these groups to safely perform their jobs. PMID- 12546236 TI - An outbreak of pneumococcal pneumonia among military personnel at high risk: control by low-dose azithromycin postexposure chemoprophylaxis. AB - In the winter of 1998-1999 an outbreak of pneumococcal pneumonia occurred among Ranger students undergoing high-intensity training. Thirty pneumonia cases (attack rate = 12.6%) were identified among a group of 239 students. Eighteen students were hospitalized; Streptococcus pneumoniae-positive cultures were detected in 11 (61.1%) of these 18 hospitalized cases. Pneumococci were also identified in throat swabs of 30 (13.6%) of 221 nonhospitalized students surveyed. Serum antipneumolysin seroconversions were detected in 30 (18.3%) of 164 students tested. An association between development of serum antipneumolysin antibody and pneumococcal pharyngeal carriage/colonization was found. Of 30 seroconverters, eight (26.7%) had S. pneumoniae-positive cultures compared with only 17 (12.7%) of 134 nonseroconverters (relative risks = 2.02, 95% confidence interval = 1.02-4.02, p = 0.05). The outbreak was controlled by administrating lowdose, oral azithromycin prophylaxis (250 mg weekly for 2 weeks) and was associated with a 69% reduction in pneumococcal carriage and a 94% reduction in pneumonia rates. PMID- 12546238 TI - Reducing unintended pregnancy in young, single active duty women in an overseas environment. AB - Unintended pregnancies in the United States negatively impact families, communities, and society as a whole. In the military, the ramifications can encroach upon mission accomplishment. In operational and overseas environments, the consequences are amplified. U.S. Naval Hospital, Sigonella, Italy became involved in a multidisciplinary effort (Choices) to reduce unplanned pregnancies. A retrospective analysis of 173 unmarried active duty women (< 27 years old) revealed that those who did not attend Choices were three times more likely to have an unplanned pregnancy. A bivariate correlation was used to test the relationship. The correlations between attendance at Choices and unplanned pregnancies was, r = -0.327, p = 0.000. A chi2 test revealed the difference in the frequency of unplanned pregnancies between those who attended Choices and those who did not with chi2 (1) = 17.86, p < 0.001 was systematic and real. This multidisciplinary approach shows promise in reducing unplanned pregnancies, thus ensuring readiness and mission accomplishment. PMID- 12546239 TI - The low impact of screening electrocardiograms in healthy individuals: a prospective study and review of the literature. AB - OBJECTIVE: To determine how often screening electrocardiograms (EKGs), which are required by military regulation, change management or disclose cardiac disease in healthy people. METHODS: A total of 1,718 consecutive EKG interpretations, whether or not the screening EKG led to further testing, disclosed serious or potentially serious cardiac disease, or changed management or disposition of the patient, were prospectively recorded. RESULTS: Thirty-four percent of the screening EKGs was abnormal. Seven (0.67%) of these abnormal EKGs altered management by leading to further consultation or testing. No screening EKG disclosed serious underlying cardiac disease. Two cases of potentially serious cardiac disease were discovered by the screening EKGs. At an average follow-up time of 34 months, neither of these two patients has developed serious cardiac disease. Our findings are similar to studies of other populations. CONCLUSION: Screening EKGs rarely caused a change in management. No adverse outcome would have occurred nor would serious cardiac disease have been missed if the routine screening EKG were not performed. The military should consider abandoning practice of performing screening EKGs in young, healthy individuals. PMID- 12546240 TI - Management of a multinational mass fatality incident in Kaprun, Austria: a forensic medical perspective. AB - The Armed Forces Regional Medical Examiner in Europe is responsible for medico legal death investigations for the U.S. military in 89 countries in Europe, Southwest Asia, and Africa. The Disaster Mortuary Affairs Response Team (DMART) is a rapidly deployable, experienced, multidisciplinary team patterned after the regional U.S. civilian Disaster Mortuary Response Teams. One of the first major challenges since the DMART's inception occurred on November 11, 2000. A fire in a funicular servicing a ski resort claimed 155 victims in a mountain tunnel in Kaprun, Austria. The DMART deployed to assist the Austrian authorities in the rapid recovery, examination, identification, and return of the remains. The DMART's considerable forensic expertise and inherent spirit of cooperation were instrumental in the successful management of the disaster, resulting in the positive identification and release of all 155 remains within 2 weeks of the incident. PMID- 12546241 TI - Resistance patterns of Mycobacterium tuberculosis complex isolates in the Turkish Army from 1998 to 2000. AB - The aim of this retrospective study was to investigate susceptibility rates of Mycobacterium tuberculosis complex (MTBC) isolates against streptomycin, rifampicin, isoniazid, and ethambutol between January 1998 and December 2000 in the Turkish Army. Specimens collected from patients were cultured both conventionally and radiometrically. Differentiation of MTBC bacteria from Mycobacteria other than tuberculosis bacilli was made by the BACTEC p-nitro-alpha acetyl-amino-beta-hydroxypropiophenone test. Susceptibility testing of MTBC isolates was performed using the BACTEC radiometric susceptibility assay for mycobacteria. Most of the specimens originated from respiratory system. A total of 98 isolates in 1998, 123 isolates in 1999, and 84 isolates in 2000 were obtained and identified as MTBC using the radiometric BACTEC TB460 system. Initial resistance was most frequent to isoniazid followed by ethambutol, streptomycin , and rifampicin in this study period. The differences between resistance rates were not statistically significant on an annual basis. None of these isolates was resistant to all four antimycobacterial agents. Although resistance rates of our isolates were not as high as previously reported by some authors from Turkey and there was no significant difference between the annual susceptibility rates, routine screening of antituberculosis drug susceptibility should be continued to control the resistance development and its spread. PMID- 12546242 TI - Allogeneic hematopoietic stem cell transplantation in the active duty military population: 1987-2001. AB - Wilford Hall Medical Center at Lackland Air Force Base is the only military medical center in the United States at which allogeneic hematopoietic stem cell transplants (HSCT) are performed. Through May 2001, 367 patients, including 163 active duty service members, have received allogeneic HSCT from related donors. We performed a retrospective review of the outcomes of allogeneic HSCT in active duty members to determine what proportion was returned to full-time military service after HSCT and to delineate the factors associated with successful return to military service. Twenty-one percent of service members who received allogeneic HSCT were returned to active duty at some point after treatment. Age, sex, branch of service, and rank had no significant influence on return to duty status. When adjusted for age and rank, the only significant predictor of successful return to active duty was a diagnosis of chronic myelogenous leukemia. Although individuals with good risk diseases such as aplastic anemia, myelodysplastic syndrome, paroxymal nocturnal hemoglobinuria, and acute myelogenous leukemia in first complete remission have also been returned active duty after HSCT, these diagnoses were not statistically predictive of a return to active duty. We conclude that it may be possible to predict which service members will return to active duty after allogeneic HSCT based on the disease for which HSCT is being performed. Most importantly, active duty service members need not be a priori medically retired after HSCT, and each return to duty decision must be individualized. PMID- 12546243 TI - Navy and Marine Corps active duty mortality patterns for 1995 to 1999. AB - The authors analyze all Navy and Marine Corps active duty deaths from January 1, 1995 through December 31, 1999 (Navy, N=1231; Marine Corps, N=701). Data were obtained from official Navy and Marine Corps sources, including the Report of Casualty (DD form 1300) and the Navy Personnel Casualty Report (Control Symbol NMPC 1770-4) or the Marine Corps Personnel Casualty Report (MC-3040-02), as appropriate. Overall fatality rates were 68.2 per 100,000 active duty Navy personnel and 84.2 for active duty Marine Corps personnel. Rates were generally lower than those noted in previous studies and lower than comparable civilian groups. The officer fatality rates were strongly affected by aircraft mishap related deaths. The only subgroup displaying higher rates than their civilian counterparts was mishap-related deaths for enlisted Marines age 17 to 24 years old. PMID- 12546244 TI - Avulsion of the common hamstring tendon origin in an active duty airman. AB - Hamstring injuries are common in active athletic populations, such as military service members. Ruptures of the hamstring origin from the ischial tuberosity are rare injuries and missed if not considered in the differential diagnosis of ischial pain. Unlike other hamstring injuries, complete hamstring avulsions must be treated surgically. Results of untreated hamstring avulsions are poor. The purpose of this article is to describe the case of an active duty airman who presented for an unrelated complaint and was discovered to have a 5-week-old hamstring avulsion. Surgical repair of the hamstring avulsion 6 weeks after injury yielded an excellent result and return to full duty. Hamstring avulsions recognized early by history and physical examination and diagnostic imaging permits early and effective treatment. Early surgical repair of the tendon to bone can result in return to full duty. PMID- 12546245 TI - Psychiatric medevacs during a 6-month aircraft carrier battle group deployment to the Persian Gulf: a Navy Force Health Protection preliminary report. AB - When a U.S. Navy Aircraft Carrier battle group deploys overseas, the aircraft carrier's medical department is responsible for the medical needs of over 12,000 personnel with their indigenous developmental, stress, family, alcohol, drug, and interpersonal and intrapersonal relationship difficulties. This article reviews the effectiveness of having a U.S. Navy clinical psychologist and a psychiatric technician onboard the USS Carl Vinson, the flag ship of Vinson's battle group, during this battle group's 1998/1999 Persian Gulf deployment (i.e., Western Pacific Deployment). Importantly, these two individuals reported to the USS Vinson as permanent members of the ship's company. The clinical psychologist logged 448 individual outpatient-care consults and 79 individual consults with sailors who had a history of overusing or abusing alcohol. Additionally, nine sailors with acute disabling psychiatric diagnoses were hospitalized on the ship's medical ward, and four sailors were medically evacuated (medevaced), by fixed wing aircraft, from USS Vinson to a Navy Hospital in the United States for definitive evaluation, treatment, and disposition. These four medevacs were less than the number of medevacs from two previous Aircraft Carrier Battle Group Persian Gulf deployments. Importantly, these two previous WESTPAC deployments were made without having a clinical psychologist as a full-time member of the respective aircraft carrier's medical department. Providing clinical psychology/mental health services at the "tip of the spear" is an effective, beneficial, and cost-saving landmark improvement in providing quality medical care to the fleet. PMID- 12546246 TI - Evaluation of stress-related anterior lower leg pain with magnetic resonance imaging and intracompartmental pressure measurement. AB - The purpose of this work was to evaluate stress-related anterior lower leg pain with clinical examination, magnetic resonance imaging, and measurement of anterior tibial compartment pressure findings. All medical data were gathered from 24 conscripts with stress-related anterior lower leg pain. Twenty exhibited bilateral symptoms. In 22 of the 44 cases, the intracompartmental pressure was pathological. Symptoms were exhibited for longer periods by patients with chronic exertional compartment syndrome (CECS) than by other patients (p < 0.01). At rest, magnetic resonance imaging revealed no abnormal findings in the soft tissues of the legs but showed bone abnormalities in 35 symptomatic legs. Thirty three exhibited bone stress injuries, and two exhibited leg traction periostitis. On magnetic resonance imaging, there was no difference in bone findings between patients with and without CECS (p > 0.05). Stress-related anterior lower leg pain can be related to CECS, bone stress injury, and traction periostitis. Clinical diagnosis is unreliable. CECS and bone stress injury or traction periostitis can occur separately or together. PMID- 12546247 TI - Removal of parasitic protozoa from water using a mobile water filtration apparatus intended for field use by military or emergency personnel. AB - Approximately 3,200 L of water were inoculated with Cryptosporidium oocysts and Giardia cysts. The contaminated water was filtered through a Berkefeld Filter, type TWA 3, as used by the Norwegian army. Nine 10-L samples of filtered water and four 10-L control samples of unfiltered water were analyzed for Cryptosporidium oocysts and Giardia cysts. On average, 1,920 Giardia cysts and 2,735 Cryptosporidium oocysts per 10 L were detected in the unfiltered water. Between 0 and 2 Giardia cysts and between 2 and 47 Cryptosporidium oocysts were detected in the filtered 10-L samples. Percentage removal of parasites was between 99.8 and > 99.95% for Giardia (2.98 to > 3.28 log removal) and between 98.3 and 99.9% (1.76-3.14 log removal) for Cryptosporidium. The Berkefeld filter TWA 3 complies with published recommendations for municipal water supply for removal of Giardia but does not always comply with such regulations for Cryptosporidium. PMID- 12546248 TI - Health assessment of U.S. Army Rangers. AB - PURPOSE: Assess the health behaviors of U.S. Army Rangers, including their diet and physical activity patterns, and their use of alcohol, tobacco, and nutritional supplements. METHODS: Army Rangers (N=38) completed selected self report and food frequency questionnaires, and anthropometric measures (height, weight, circumferences) were taken. RESULTS: Dietary patterns were collected from questionnaires that asked respondents to report the frequency of usual consumption from a list of foods over a period of 1 year. The data collected indicated a diet high in fat (38.0% of energy) and low in carbohydrates (41.9%). Protein intake (17.9% of energy) was consistent with recommendations for endurance and strength training athletes. Hours engaged in physical activity averaged 12 per week. Overall, 76% of the volunteers reported using alcohol on a regular basis, and 52.6% used some form of tobacco: 50% reported using either snuff or chewing tobacco. The use of supplements was high with 13% taking creatine and ephedrine products on a regular basis. CONCLUSION: Although physical activity patterns indicated a high level of activity, dietary and other health behaviors, such as smoking, alcohol use, and patterns of supplement use, suggest that health education programs may be important in this select Army population. PMID- 12546249 TI - The physiological and psychological effects of combat ration feeding during a 12 day training exercise in the tropics. AB - The health and psychological effects of combat ration pack (CRP) feeding during 12 days of military training in a tropical environment were investigated. Three groups of air defence guards (ADGs) received either: freshly prepared foods (fresh group, 15 mega joule (MJ), 3,600 kcal, N=13), full CRP (15 MJ, 3,600 kcal, N=10), or one-half CRP (7.5 MJ, 1,800 kcal, N=10). Underconsumption by the full CRP group resulted in CRP groups experiencing similar weight loss, protein catabolism, and immune suppression (both cell-mediated and humoral), whereas the fresh group maintained their weight and protein balance and cell-mediated immune status. CRP groups reported greater fatigue than the fresh group. All ADGs experienced poor sleep quality and declining folate and iron status. ADGs drank insufficient water to prevent dehydration. In the medium term, ADGs were able to adapt to restricted food consumption and poor sleep quality with no decrement in physical fitness or cognition. PMID- 12546250 TI - Prevalence and characteristics of trauma and post-traumatic stress symptoms in operational members of the South African National Defence Force. AB - The prevalence of traumatic event exposure and post-traumatic stress disorder (PTSD) were surveyed in a cohort of 198 full-time operational members of the South African National Defence Force stationed in their "home" unit between deployments. Approximately 90% of members reported having experienced or witnessed trauma in their lifetime (mean number of traumatic events = 4.3 +/- 3.2), whereas 51% reported having inflicted trauma. Twenty-six percent met diagnostic criteria for PTSD on self-report with approximately 29% with PTSD also meeting diagnostic criteria for depression. Few members, however, sought help. PTSD symptom severity was best predicted by trauma type (exposure to physical assault and infliction of life-threatening injury). These findings highlight the high rates of exposure to multiple, noncombat-related trauma in military personnel, the potentially high rates of PTSD, and the role of inflicted trauma as an additional risk factor for PTSD. PMID- 12546251 TI - Collapse from exertional heat illness: implications and subsequent decisions. AB - The implications of the collapse of a soldier early in an exercise from exertional heat illness (EHI) are considered. Such soldiers may be at risk from a genetic predisposition. Malignant hyperthermia (MH) and isolated and improbable cases of EHI may be just two different expressions of the same mutated gene sequence. The genetics of MH are complex and present knowledge is incomplete. The use of the in vitro contraction test (IVCT) on cases of EHI, in addition to its proven role in MH, would be helpful in examining the relationship between MH and EHI. It has been shown that some soldiers collapsing with EHI may have subsequent positive IVCTs. The test, however, sometimes produces false positives and, in addition, a positive result could be a consequence of a heat insult rather than an antecedent. Further studies to establish the incidence of positive IVCTs in relatives of EHI probands, and thus test for heritability, are required. There is, at the moment, only one example of proven MH and proven EHI occurring in the same individual. DNA from a 12-year-old boy who suffered MH and later died from the EHI and from his relatives showed relevant mutations as did the DNA of two of three soldiers who survived EHI. Hajj pilgrims, who collapse with heat illness, do not show such mutations, but the etiology is different. The sedentary pilgrims succumbed to a very high external ambient temperature, the active soldiers to a huge output of internal metabolic heat. Only eventual advances in defining the genetics of MH and EHI will resolve the present confusion of the relationship between the two conditions. Meanwhile, there is a need to bypass considerations of the etiology of EHI and to identify the vulnerable and handicapped soldier by exposure after an interval of time to one or more exercise tolerance tests. PMID- 12546252 TI - A 41-year-old active duty U.S. Air Force enlisted man with a 3-week history of fevers, arthralgias, and myalgias. AB - A 41-year-old active duty male was transferred to Walter Reed Army Medical Center in Washington, DC, for further evaluation of fever, rash, myalgias, arthralgias, and respiratory failure. An extensive evaluation with input from numerous subspecialties of medicine was performed. The patient was eventually diagnosed with adult Still's disease, which is a diagnosis of exclusion. This case illustrates the importance of having a broad differential diagnosis when evaluating a patient with fever of unknown origin, with emphasis on the approach to young, active duty military personnel. PMID- 12546253 TI - Capitated assertive community treatment program savings: system implications. AB - In a California county of one million people, 500 clients, 4% of all those served in 1994, were found to use 38% of publicly funded mental health services. A controlled experiment was designed to test whether a capitated Assertive Community Treatment (ACT) program could produce outcomes that were equivalent or better than "usual services" for a subset of very-high-utilizing clients while reducing costs. Results showed that major challenges in using an ACT program for cost reduction were successfully met. Costs over all 4 years were substantially lower for the experimental group than for a randomly assigned comparison group. PMID- 12546254 TI - Practice guidelines in mental health and addiction services: contributions from the American College of Mental Health Administration. AB - Practice guidelines have proliferated in mental health and addiction services; more than 40 organizations have developed guidelines in the field. However, much confusion, controversy, and contextual issues remain, particularly regarding effective dissemination and the clinical, financial, political, and ethical issues that emerge when evidence-based behavioral health practice guidelines are implemented in systems of care. The American College of Mental Health Administration (ACMHA) focused on these problems in their 1999 and 2000 Santa Fe Summits and produced a number of specific outcomes that contribute to thinking in the field. These include the following: (a) a taxonomy of building blocks for informed decision-making in behavioral health assessment and treatment; (b) a paradigm for the development of practice guidelines; (c) characteristics of a good practice guideline; (d) strategies for disseminating and implementing practice guidelines; and (e) areas in need of future research. PMID- 12546255 TI - Symptom severity, amount of treatment, and 1-year outcomes among dual diagnosis patients. AB - This study reports on associations among symptom severity, amount of treatment, and 1-year outcomes in a national sample of 8,622 dual diagnosis patients, who were classified at treatment entry into low-, moderate-, and high-severity groups. Patients with more severe symptoms at intake had poorer 1-year outcomes. Higher severity patients did not receive adequate "doses" of care: Compared with low-severity patients, they had a shorter duration of care, although a longer duration was associated with improved outcomes; they also were less likely to receive outpatient substance abuse treatment, although more intensive treatment was associated with better drug outcomes. High-severity patients improved more on drug and legal outcomes, but less on psychiatric and family/social outcomes, than low-severity patients did when treatment was of longer duration or higher intensity. Dual diagnosis patients with highly severe symptoms would likely benefit from a longer episode of care that includes substance abuse and psychiatric outpatient treatment. PMID- 12546256 TI - Integrated local networks as a model for organizing mental health services. AB - Organizing services in an integrated network as a model for transforming healthcare systems is often presented as a potential remedy for service fragmentation that should enhance system efficiency. In the mental health sector, integration is also part of a diversified response to the multiple needs of the clients, particularly people with serious mental health disorders. The authors describe how the notion of integrated service networks came to serve as a model for transforming the mental health system in Quebec, and they propose a frame of reference for this notion. They also address the challenges and issues raised by this mode of service organization in the mental health sector and more generally in a context of transforming healthcare systems. PMID- 12546257 TI - Administrative support is important for a successful psychosocial rehabilitation program in a public psychiatric hospital. PMID- 12546258 TI - Asian American and Pacific Islander mental health and substance abuse agencies: organizational characteristics and service gaps. PMID- 12546259 TI - Bispectral Index in ICU: correlation with Ramsay Score on assessment of sedation level. AB - OBJECTIVE: The assessment of sedation level in critically ill patient remains a challenge for the intensivists in order to avoid over- or under-sedation phenomena. Scoring systems commonly used still show some limitation; the introduction of Bispectral Index (an EEG parameter) could bring potential advantages in monitoring sedation. According to the reports, Bispectral Index correlates with levels of sedation on the Ramsay Scale. We report our personal experience in this topic. METHODS: Twenty patients, diagnosed with chronic obstructive pulmonary disease (COPD), were involved in the study. For an optimal adaptation to respiratory prothesis, they were sedated (with Propofol by continuous infusion at an initial dose of 2 mg/kg/h, which could be modulated with steps of 0.5 mg/kg/h), in order to maintain a Ramsay score of 4-5. BIS value was continuously recorded, and manually calculated on a mean average of a minute during the measuring of Ramsay score (T0) and every 30' for 24 hours on par with Ramsay score. EEG, SpO2, non invasive arterial pressure, ventilatory module, ETCO2, FIO2 were also recorded. For the statistic analysis, Friedman test and Spearman coefficient were utilized. Values of p < 0.05 were considered significant. RESULTS: 980 observations were carried out. The variation range of Ramsay Score was between 2 and 6. BIS range varied from 34 to 98. Statistic analysis of the data obtained pointed out some significative correlations, particularly between Ramsay Score and BIS (p < 0.01), and between BIS and dosage of Propofol (p < 0.01). CONCLUSIONS: The results of the study are consistent with those found in the literature on this topic of study. In fact, this study demonstrates the utility of BIS to track levels of consciousness in ICU patients while still maintaining the use of the score systems to care for ICU patients. PMID- 12546260 TI - Diagnosis of anesthetic-induced upper airway obstruction in children using respiratory inductance plethysmography. AB - OBJECTIVE: Upper airway obstruction is the most rapid and clinically relevant cause of hypoxia during sedation and anesthesia. This study was designed to determine if respiratory inductance plethysmography (RIP) could quantify the degree of upper airway obstruction caused by induction of general anesthesia. METHODS: RIP tracings were obtained during induction of general anesthesia in healthy children. Three sets of measurements were obtained during: (1) a 3 minute control period without anesthetics, (2) 3 minutes of 50% nitrous oxide, and (3) halothane administration to complete the induction of general anesthesia. Clinical impression of upper airway obstruction (none, partial, or complete) was correlated with two separate RIP analysis techniques. RESULTS: Three hundred ninety-five breathing epochs from 20 children (ages 3-10 years) were analyzed by both phase shift and phase inversion techniques. Although both techniques had good general correlation with severity of airway obstruction, neither was sufficiently reliable for accurate prediction of severity of airway obstruction. CONCLUSIONS: We investigated two methods for analyzing RIP tracings during varying degrees of upper airway obstruction in anesthetized children. We found that neither technique was sufficiently accurate for predicting the severity of upper airway obstruction and would not be useful as a predictor of upper airway obstruction in the clinical or research settings. PMID- 12546262 TI - Learning about new anesthetics using a model driven, full human simulator. AB - OBJECTIVE: New pharmacological agents are introduced into medical practice at an ever-increasing pace. Teaching how to use new medications in the clinical setting presents educational challenges and puts patients at risk. METHODS: Patients and clinical settings in which remifentanil might provide clinical advantages over existing anesthetics were identified. A simulator curriculum was developed to demonstrate the use of remifentanil in the sample cases. The simulation was designed to highlight the clinical advantages and potential side effects of remifentanil. A screen displaying the concentrations of remifentanil in plasma and in the hypothetical effector site was developed. A simulator was modified (addition of an infusion pump and a pharmacokinetic screen display) and transported to several cities in the U.S.A. An instructor guided small groups of anesthesiologists and anesthetists through a structured program that enabled participants to observe drug effects in simulated patients. RESULTS: There were 836 participants in the remifentanil program, which was offered in 58 cities in the U.S.A. Surveys were completed by 574 anesthesiologists. There was a significant difference in comfort level for using remifentanil after the session compared to before (Chi-square, p < 0.001.) The statement: "Clinical simulation experience is a means to learn about new agents like remifentanil" was rated as "excellent" by 81% and as "good" by 19% of participants. No participant found the experience to be "not useful." CONCLUSIONS: Patient simulation is a novel method of introducing new drugs to the medical community and is perceived by anesthesia providers as a valuable addition to available teaching methods. PMID- 12546261 TI - Activated clotting time systems vary in precision and bias and are not interchangeable when following heparin management protocols during cardiopulmonary bypass. AB - OBJECTIVE: Our aim was to test the hypothesis that new activated clotting time (ACT) technology, with modifications to instruments and reagents designed to detect earlier clot formation, would be associated with more precise but lower results. A secondary objective was to evaluate the potential impact of any change in ACT measurement on heparin requirements during cardiopulmonary bypass (CPB). METHODS: We compared the precision of two newer ACT systems: Actalyke, Helena Laboratories, Beaumont, TX and Hemochron Response, International Technidyne Corporation, Edison, NJ and assessed their bias with reference to a standard ACT system (Hemochron 801, International Technidyne Corporation, Edison, NJ). Bland Altman analysis was applied to 81 duplicate samples from 22 patients undergoing CPB or percutaneous coronary interventions (PCI), covering the full clinical range of ACT values. We also estimated the change in heparin dose required to use the Actalyke rather than the Hemochron 801 results, to achieve our target ACT for CPB (480 seconds), and used a mixed model to test for significance. RESULTS: The precision of the Actalyke was superior to the Hemochron Response (mean difference of duplicates +/- 0.1% versus +/- 4.2%). There was no significant bias (p = 0.93) between the results from the standard analyzers (Hemochron 801 and Response), but the results from the modified system (Actalyke) were on average 18% lower than the Hemochron 801 (p < 0.0001). Estimated heparin requirements established that fifty percent of CPB patients would have required additional heparin (5000 to 17500 units), an average increase of 1060 units per patient (p = 0.05), if the Actalyke values were used to guide anticoagulation during CPB. CONCLUSIONS: Our results support the hypothesis that the modified technology (Actalyke) is associated with more precise but lower ACT results. We estimated these lower values would lead to increased heparin dosing during CPB. The impact of this increase on bleeding after cardiac surgery with CPB is controversial and requires further study. PMID- 12546263 TI - Monitoring of motor evoked potentials with high intensity repetitive transcranial electrical stimulation during spinal surgery. AB - OBJECTIVE: Clinical utility of high voltage repetitive transcranial electrical stimulation (TES) was investigated in 46 patients undergoing spine surgery. METHODS: During spinal surgery, motor evoked potentials (MEPs) were recorded from upper or lower limb muscles following high voltage repetitive TES of motor cortex under propofol and opioid/N2O anesthesia. RESULTS: The number of responses evoked by the double pulse stimulation was significantly higher than the single pulse stimulation. A similar finding was obtained when repetitive and single pulse stimulation was compared. Compound muscle action potentials (CMAPs) were recorded from upper and lower limbs in 4 patients with cervical spine myclopathy. The CMAP was absent on the affected side in 1 patient, which improved slightly after decompression. Radiculopathy was clinically present in 6 patients undergoing posterior lumbar decompression and fusion. No improvement of MEP was noted intraoperatively after spinal decompression and instrumentation. CONCLUSION: The findings suggest that intraoperative MEP monitoring is feasible method, however, its immediate prognostic value for adequacy of neuronal decompression and improvement requires further studies with larger patient population. PMID- 12546264 TI - Subcutaneous microdialysis for metabolic monitoring in abdominal aortic surgery. AB - Microdialysis, that is the sampling of interstitial fluid via semi-permeable tubes, has been shown to be suitable for detecting ischemic changes e.g. in brain and heart tissue. The purpose of the present study was to investigate the possibility of monitoring with subcutaneous microdialysis peri-operative metabolic sequelac of elective abdominal aortic surgery. In 22 patients microdialysis catheters were inserted subcutaneously in the lower leg as well as the shoulder serving as a reference topographic region. Lactate, pyruvate and glycerol, which indicate ischemia or reperfusion, were measured. We observed severe metabolic changes within the interstitial fluid of the lower extremity during ischemia followed by normalization during reperfusion. Despite high interstitial concentrations of lactate and glycerol indicating severe ischemia during clamping of the abdominal aorta these parameters returned to pre-operative values within 2 hrs after declamping and all patients recovered completely. OBJECTIVE: Information about the metabolic state of the lower extremity during and after infrarenal aortic aneurysm repair should modify peri-operative treatment. The aim of the study was to evaluate whether microdialysis of the subcutaneous tissue reflects metabolic changes during ischemia and reperfusion. Lactate, pyruvate and glycerol concentrations were measured in the subcutaneous tissue of the lower extremity and compared to the microdialysis measurements from shoulder subcutaneous tissue. METHOD: In 22 patients microdialysis catheters were inserted preoperatively in the subcutaneous space of the left shoulder and the left calf. Samples were taken at timed intervals before, during and after clamping of the abdominal aorta. RESULTS: The subcutaneous glycerol concentration of the calf was increased during the clamping period from initially 68 +/- 11 microM up to 182 +/- 27 microM (p < 0.05); the lactate/pyruvate (L/P) ratio was increased eightfold. After declamping these values normalized to baseline. Microdialysis measurements of the shoulder showed no prominent changes during the entire course of observation. CONCLUSIONS: Subcutaneous microdialysis was able to detect metabolic changes due to ischemia during clamping of the abdominal aorta as well as reperfusion there after. It is a suitable technique to monitor the peri-operative course of the dependent tissue after abdominal aortic vascular surgery. PMID- 12546265 TI - Pulmonary blood flow (cardiac output) and the effective lung volume determined from a short breath hold using the differential Fick method. AB - OBJECTIVES: This work attempts to show how pulmonary blood flow (Qp), cardiac output (CO(Fick)) and the lung volume of effective gas exchange (ELV) can be determined from breath-by-breath measurements of the tidal exhaled CO2 elimination V (litre/min) and the end tidal CO2 concentration P (%) using the differential Fick method. The measurements are made during steady state ventilation and when the CO2 balance in the lungs changes subsequent to a perturbation of the gas exchange conditions. METHODS: A short breath hold is used to implement such a perturbation. V and P were measured in patients on mechanical ventilation. When the end tidal CO2 values were stable, the end inspiratory pause of a single breath was prolonged 3 seconds as compared to the normal ventilation pattern. From the changes induced in P and V, Qp, CO(Fick) and ELV are obtained. RESULTS: Cardiac output values were measured in 18 patients after CABG. CO(Fick) was found to be in good agreement with the thermodilution cardiac output reference COth. (n = 51 and (COth-CO(Fick)) with Mean = -0.17 litre/ minute and SI) = 0.62 litre/minute). CONCLUSIONS: With a single breath perturbation, the differential Fick method can yield cardiopulmonary information using 2-3 breaths only and with a minimum of interference with the patient. Complete data analysis results in multiple determinations of the Qp and ELV values which improve the attainable precision. Our investigation points to the possibility to determine Qp, CO(Fick) and ELV also during spontaneous breathing, by using the natural tidal variations of V and P. PMID- 12546266 TI - Who is conducting the deadly quartet? PMID- 12546267 TI - Emphysema in nonsmokers: alpha 1-antitrypsin deficiency and other causes. AB - Nonsmokers with signs of emphysema at an earlier age than is typical for emphysema deserve a workup for one of the less common causes of emphysema, which include alpha 1-antitrypsin deficiency, connective tissue diseases, hypocomplementemic urticarial vasculitis syndrome, intravenous drug use, human immunodeficiency virus infection, and several rare metabolic disorders. PMID- 12546268 TI - Update on the diagnosis and treatment of human papillomavirus infection. AB - Human papillomaviruses (HPVs) are associated with a spectrum of diseases, ranging from common warts to invasive carcinoma of the genital tract. The clinical manifestations of HPV infection depend on the viral subtype, the immune status of the patient, and environmental co-carcinogens. Infection with HPV is often asymptomatic, which makes viral detection challenging. Current therapies do not reliably eradicate HPV infection, and benign genital warts and genital tract intraepithelial neoplasia often recur after treatment. We discuss the pathogenesis, clinical manifestations, detection, and treatment of HPV infections of the anogenital tract. PMID- 12546269 TI - Risedronate prevents hip fractures, but who should get therapy? AB - The Hip Intervention Program (HIP) trial establishes that risedronate (Actonel) prevents hip fracture in elderly women with osteoporosis. However, the drug had no statistically significant effect on hip fracture risk in elderly women in whom bone density status was not known. Patients should be selected for bisphosphonate therapy on the basis of low bone density. A history of vertebral fractures increases the risk for hip fractures. PMID- 12546270 TI - Gallbladder disease: an update on diagnosis and treatment. AB - This paper reviews the clinical presentation of gallstone disease, acalculous cholecystitis, biliary dyskinesia, and gallbladder cancer, as well as how to make best use of current diagnostic and treatment methods, particularly ultrasonography, cholescintigraphy, laparoscopic cholecystectomy, and endoscopic retrograde cholangiopancreatography. PMID- 12546271 TI - A truly deadly quartet: obesity, hypertension, hypertriglyceridemia, and hyperinsulinemia. AB - The National Cholesterol Education Program recognizes the importance of the metabolic syndrome and has published guidelines for its diagnosis. Weight loss, physical activity, and treatment of the individual risk factors constitute the main strategies for treatment. For now, the goals and methods of treating hypertension and dyslipidemia are the same in people with the metabolic syndrome as in the general population. Thiazolidinedione drugs increase insulin sensitivity, but their use in the metabolic syndrome is only speculative at present. We recommend they be used only as indicated to treat diabetes mellitus. PMID- 12546272 TI - The metabolic syndrome: a tug-of-war with no winner. AB - The metabolic syndrome may be viewed as a state of insulin-counterregulatory overdrive: counterregulatory hormones and fatty acids chronically duel with insulin, causing a cascade of biochemical interactions resulting in insulin resistance, hypertension, and dyslipidemia. Even before beta cells fail and type 2 diabetes ensues, the deadly quartet is quietly rehearsing. PMID- 12546273 TI - HIV update 2002: delaying treatment to curb rising resistance. AB - The recommendation for HIV antiretroviral therapy is changing from "treat early, treat hard" to "treat a little later, treat hard" as more toxicities of the drugs are recognized. Drug-resistant HIV is becoming a serious problem, even in treatment-naive patients. For HIV patients with organ failure, transplantation is increasingly an option. PMID- 12546274 TI - Which children and adults should receive the chickenpox vaccine? AB - Chickenpox vaccine is safe and effective and elicits long-standing immunity. Primary care physicians caring for children and adults should be aware of its benefits. PMID- 12546275 TI - Alteration of endothelins: a common pathogenetic mechanism in chronic diabetic complications. AB - Endothelin (ET) peptides perform several physiological, vascular, and nonvascular functions and are widely distributed in a number of tissues. They are altered in several disease processes including diabetes. Alteration of ETs have been demonstrated in organs of chronic diabetic complications in both experimental and clinical studies. The majority of the effects of ET alteration in diabetes are due to altered vascular function. Furthermore, ET antagonists have been shown to prevent structural and functional changes induced by diabetes in animal models. This review discusses the contribution of ETs in the pathogenesis and the potential role of ET antagonism in the treatment of chronic diabetic complications. PMID- 12546276 TI - Effect of acipimox on plasma lipids and glucose/insulin in pregnant rats. AB - To determine how a reduction in maternal hypertriglyceridemia during late pregnancy may affect glucose/insulin relationships, pregnant and virgin rats were orally treated with acipimox, a potent antilipolytic agent. In 20-day pregnant rats receiving 80 mg of acipimox, plasma triglycerides (TG), free fatty acids (FFA), and glycerol decreased more than in virgin rats shortly after the drug (up to 7 hours), when compared with animals treated with distilled water, whereas plasma glucose level was unaffected by the treatment in either group of rats. When acipimox was given every 12 hours from day 17 to day 20 of pregnancy, plasma TG, FFA, and glycerol levels progressively increased, whereas they either decreased or did not change in virgin rats receiving the same treatment, with no effect in plasma glucose levels in either group. Fetal body weight was lower than in controls in 20-day pregnant rats that received acipimox for 3 days. On day 20 of pregnancy, 3 hours after receiving acipimox or distilled water, rats received a 2 g glucose/kg oral load and it was found that the change in plasma glucose was similar in both groups, whereas the increase in plasma insulin was greater in pregnant rats treated with acipimox. However, no difference was found in either variable after the oral glucose load in virgin rats receiving acipimox or distilled water. No differences in plasma glucose levels were found after intravenous (i.v.) administration of insulin in pregnant rats treated or not treated with acipimox. In conclusion, present results show that administration of acipimox during the last days of gestation inhibited lipolysis and decreased fetal weight. Over a short period of time, in pregnant rats, reductions of plasma FFA and TG after acipimox treatment improved the glucose-induced insulin release, but did not seem to have any effect in peripheral insulin resistance. PMID- 12546277 TI - C-peptide prevents hippocampal apoptosis in type 1 diabetes. AB - To explore mechanisms underlying central nervous system (CNS) complications in diabetes, we examined hippocampal neuronal apoptosis and loss, and the effect of C-peptide replacement in type 1 diabetic BB/W rats. Apoptosis was demonstrated after 8 months of diabetes, by DNA fragmentation, increased number of apoptotic cells, and an elevated ratio of Bax/Bcl-xL, accompanied by reduced neuronal density in the hippocampus. No apoptotic activity was detected and neuronal density was unchanged in 2-month diabetic hippocampus, whereas insulin-like growth factor (IGF) activities were impaired. In type 1 diabetic BB/W rats replaced with C-peptide, no TdT-mediated dUTP nick-end labeling (TUNEL)-positive cells were shown and DNA laddering was not evident in hippocampus at either 2 or 8 months. C-peptide administration prevented the preceding perturbation of IGF expression and reduced the elevated ratio of Bax/Bcl-xL. Our data suggest that type 1 diabetes causes a duration-dependent programmed cell death of the hippocampus, which is partially prevented by C-peptide. PMID- 12546278 TI - The role of reactive oxygen species in diabetes-induced anomalies in embryos of Cohen diabetic rats. AB - The role of the antioxidant defense mechanism in diabetes-induced anomalies was studied in the Cohen diabetes-sensitive (CDs) and -resistant (CDr) rats, a genetic model of nutritionally induced type 2 diabetes mellitus. Embryos, 12.5 day-old, of CDs and CDr rats fed regular diet (RD) or a diabetogenic high-sucrose diet (HSD) were monitored for growth retardation and congenital anomalies. Activity of superoxide dismutase (SOD) and catalase-like enzymes and levels of ascorbic acid (AA), uric acid (UA), and dehydroascorbic acid (DHAA) were measured in embryonic homogenates. When fed RD, CDs rats had a decreased rate of pregnancy, and an increased embryonic resorption. CDs embryos were smaller than CDr embryos; 46% were maldeveloped and 7% exhibited neural tube defects (NTDs). When fed HSD, rate of pregnancy was reduced, resorption rate was greatly increased (56%; P < .001), 47.6% of the embryos were retrieved without heart beats, and 27% exhibited NTD. In contrast, all the CDr embryos were normal when fed RD or HSD. Activity of SOD and catalase was not different in embryos of CDs and CDr rats fed RD. When fed HSD, levels of AA were significantly reduced, the ratio DHAA/AA was significantly increased, and SOD activity was not sufficiently increased when compared to embryos of CDr. The reduced fertility of the CDs rats, the growth retardation, and NTD seem to be genetically determined. Maternal hyperglycemia seems to result in environmentally induced embryonic oxidative stress, resulting in further embryonic damage. PMID- 12546279 TI - Effect of mild hypoinsulinemia on renal hypertrophy: growth hormone/insulin-like growth factor I system in mild streptozotocin diabetes. AB - The metabolic aberrations associated with diabetes mellitus profoundly alter the growth hormone/insulin-like growth factor I (GH/IGF-I) system. In severe experimental diabetes, serum IGF-I level is reduced, reflecting altered hepatic expression. On the other hand, increased levels of kidney IGF-I have been implicated in the development of diabetic kidney disease. This study aimed to examine the effect of mild experimental diabetes with hypoinsulinemia on both the systemic and renal GH/IGF-I systems in a low-dose streptozotocin (STZ)-induced diabetic rat. Diabetic animals with mild hypoinsulinemia developed renal hyperfiltration within 3 days of diabetes, whereas the renal size increased significantly only between 30 and 48 days of diabetes. Plasma GH levels were unchanged during the entire course of the study, but a decrease in serum IGF-I, IGF-binding protein 3 (IGFBP-3), and IGF-binding protein 4 (IGFBP-4) occurred after 10, 30, and 48 days. Kidney IGF-I and IGF-binding protein 1 (IGFBP-1) mRNA expression increased after 10 and 30 days of diabetes. A significant increase in kidney IGFBP-1/2, IGFBP-3, and IGFBP-4 proteins was seen after 48 days of diabetes. A positive correlations was found between renal growth and insulin/glucose ratio (r=.57), kidney IGF-I (r=.57), IGFBP-1 mRNA (r=.43), IGFBP 1/2 (r=.41), and IGFBP-4 levels (r=.40). These results demonstrate hyperfiltration within 3 days of diabetes and a similar response in the IGF-I system in mildly and severely hypoinsulinemic rats; however, renomegaly develops slower in mildly diabetic rats at least partly due to delayed changes in the renal IGF and IGF BPs. PMID- 12546280 TI - Access to care: remembering old lessons. PMID- 12546281 TI - Medigap premiums and Medicare HMO enrollment. AB - OBJECTIVE: Markets for Medicare HMOs (health maintenance organizations) and supplemental Medicare coverage are often treated separately in existing literature. Yet because managed care plans and Medigap plans both cover services not covered by basic Medicare, these markets are clearly interrelated. We examine the extent to which Medigap premiums affect the likelihood of the elderly joining managed care plans. DATA SOURCES: The analysis is based on a sample of Medicare beneficiaries drawn from the 1996-1997 Community Tracking Study (CTS) Household Survey by the Center for Studying Health System Change. Respondents span 56 different CTS sites from 30 different states. Measures of premiums for privately purchased Medigap policies were collected from a survey of large insurers serving this market. Data for individual, market, and HMO characteristics were collected from the CTS, InterStudy, and HCFA (Health Care Financing Administration). STUDY DESIGN: Our analysis uses a reduced-form logit model to estimate the probability of Medicare HMO participation as a function of Medigap premiums controlling for other market- and individual-level characteristics. The logit coefficients were then used to simulate changes in Medicare participation in response to changes in Medigap premiums. PRINCIPAL FINDINGS: We found that Medigap premiums vary considerably among the geographic markets included in our sample. Measures of premiums from different insurers and for different types of Medigap policies were generally highly correlated across markets. Our models consistently indicate a strong positive relationship between Medigap premiums and HMO participation. This result is robust across several specifications. Simulations suggest that a one standard deviation increase in Medigap premiums would increase HMO participation by more than 8 percentage points. CONCLUSIONS: This research provides strong evidence that Medigap premiums have a significant effect on seniors' participation in Medicare HMOs. Policy initiatives aimed at lowering Medigap premiums will likely discourage enrollment in Medicare HMOs, holding other factors constant. Although the Medigap premiums are just one factor affecting the future penetration rate of Medicare HMOs, they are an important driver of HMO enrollment and should be considered carefully when creating policy related to seniors' supplemental coverage. Similarly, our results imply that reforms to the Medicare HMO market would influence the demand for Medigap policies. PMID- 12546283 TI - Racial differences in predictors of dental care use. AB - OBJECTIVE: To test five hypotheses that non-Hispanic African Americans (AAs) and non-Hispanic whites (NHWs) differ in responsiveness to new dental symptoms by seeking dental care, and differ in certain predictors of dental care utilization. DATA SOURCES/STUDY SETTING: Florida Dental Care Study, comprising AAs and NHWs 45 years old or older, who had at least one tooth, and who lived in north Florida. STUDY DESIGN: We used a prospective cohort design. The key outcome of interest was whether dental care was received in a given six-month period, after adjusting for the presence of certain time-varying and fixed characteristics. DATA COLLECTION/EXTRACTION METHODS: In-person interviews were conducted at baseline and 24 months after baseline, with six-monthly telephone interviews in between. PRINCIPAL FINDINGS: African Americans were less likely to seek dental care during follow-up, with or without adjusting for key predisposing, enabling, and oral health need characteristics. African Americans were more likely to be problem oriented dental attenders, to be unable to pay an unexpected $500 dental bill, and to report postbaseline dental problems. However, the effect of certain postbaseline dental signs and symptoms on postbaseline dental care use differed between AAs and NHWs. Although financial circumstance was predictive for both groups, it was more salient for NHWs in separate NHW and AA regressions. Frustration with past dental care, propensity to use a homemade remedy, and dental insurance were significant predictors among AAs, but not among NHWs. The NHWs were much more likely to have sought care for preventive reasons. CONCLUSIONS: Racial differences in responsiveness to new dental symptoms by seeking dental care were evident, as were differences in other predictors of dental care utilization. These differences may contribute to racial disparities in oral health. PMID- 12546282 TI - Health insurance and mammography: would a Medicare buy-in take us to universal screening? AB - OBJECTIVE: To determine whether health insurance expansions via a Medicare buy-in might plausibly increase mammography screening rates among women aged 50-64. DATA SOURCES: Two waves of the Health and Retirement Study (HRS) (1994, 1996). STUDY DESIGN: A longitudinal study with most explanatory variables measured at the second wave of HRS (1994); receipt of mammography, number of physician visits, and breast self exam (BSE) were measured at the third wave (1996). DATA EXTRACTION: Our sample included women aged 50-62 in 1994 who answered the second and third HRS interview (n = 4,583). PRINCIPAL FINDINGS: From 1994 to 1996, 72.7 percent of women received a mammogram. Being insured increased mammography in both unadjusted and adjusted analyses. A simulation of universal insurance coverage in this age group increased mammography rates only to 75-79 percent from the observed 72.7 percent. When we accounted for potential endogeneity of physician visits and BSE to mammography, physician visits remained a strong predictor of mammography but BSE did not. CONCLUSION: Even in the presence of universal coverage and very optimistic scenarios regarding the effect of insurance on mammography for newly insured women, mammography rates would only increase a small amount and gaps in screening would remain. Thus, a Medicare buy in could be expected to have a small impact on mammography screening rates. PMID- 12546284 TI - Usual source of care in preventive service use: a regular doctor versus a regular site. AB - OBJECTIVE: To compare the effects of having a regular doctor and having a regular site on five preventive services, controlling for the endogeneity of having a usual source of care. DATA SOURCE: The Medical Expenditure Panel Survey 1996 conducted by the Agency for Healthcare Research and Quality and the National Center for Health Statistics. STUDY DESIGN: Mammograms, pap smears, blood pressure checkups, cholesterol level checkups, and flu shots were examined. A modified behavioral model framework was presented, which controlled for the endogeneity of having a usual source of care. Based on this framework, a two equation empirical model was established to predict the probabilities of having a regular doctor and having a regular site, and use of each type of preventive service. PRINCIPAL FINDINGS: Having a regular doctor was found to have a greater impact than having a regular site on discretional preventive services, such as blood pressure and cholesterol level checkups. No statistically significant differences were found between the effects a having a regular doctor and having a regular site on the use of flu shots, pap smears, and mammograms. Among the five preventive services, having a usual source of care had the greatest impact on cholesterol level checkups and pap smears. CONCLUSIONS: Promoting a stable physician-patient relationship can improve patients' timely receipt of clinical prevention. For certain preventive services, having a regular doctor is more effective than having a regular site. PMID- 12546285 TI - The lack of effect of market structure on hospice use. AB - OBJECTIVE: To describe the relative importance of health care market structure and county-level demographics in determining rates of hospice use. DATA SOURCES: Medicare claims data for a cohort of elderly patients newly diagnosed with lung cancer, colon cancer, stroke, or heart attack in 1993, followed for up to five years, and linked to Census and Area Resource File data. STUDY DESIGN: Variation between markets in rates of hospice use by patients with serious illness was examined after taking into account differences in individual-level data using hierarchical linear models. The relative explanatory power of market-level structure and local demographic variables was compared. DATA COLLECTION METHODS: The cohort was defined within the Medicare hospital claims data using validated algorithms to detect incident cases of disease with a three-year lookback. Use of hospice was determined by linkage at an individual level to the Standard Analytic Files for Hospice through 1997. Individual-level data was linked to the Area Resource File using county identifiers present in the Medicare claims. PRINCIPAL FINDINGS: There is substantial variation in hospice use across markets. This variation is not explained by differences in the major components of health care infrastructure: the availability of hospital, nursing home, or skilled nursing facilities, nor by the availability of HMOs, doctors, or generalists. CONCLUSIONS: Intercounty heterogeneity in hospice use is substantial, and may not be related to the set-up of the medical care system. The important local factors may be local preferences, differences in the particular mix of services provided by local hospices, or differences in community leadership on end of life-issues; many of these differences may be amenable to educational efforts. PMID- 12546286 TI - Error reduction and performance improvement in the emergency department through formal teamwork training: evaluation results of the MedTeams project. AB - OBJECTIVE: To evaluate the effectiveness of training and institutionalizing teamwork behaviors, drawn from aviation crew resource management (CRM) programs, on emergency department (ED) staff organized into caregiver teams. STUDY SETTING: Nine teaching and community hospital EDs. STUDY DESIGN: A prospective multicenter evaluation using a quasi-experimental, untreated control group design with one pretest and two posttests of the Emergency Team Coordination Course (ETCC). The experimental group, comprised of 684 physicians, nurses, and technicians, received the ETCC and implemented formal teamwork structures and processes. Assessments occurred prior to training, and at intervals of four and eight months after training. Three outcome constructs were evaluated: team behavior, ED performance, and attitudes and opinions. Trained observers rated ED staff team behaviors and made observations of clinical errors, a measure of ED performance. Staff and patients in the EDs completed surveys measuring attitudes and opinions. DATA COLLECTION: Hospital EDs were the units of analysis for the seven outcome measures. Prior to aggregating data at the hospital level, scale properties of surveys and event-related observations were evaluated at the respondent or case level. PRINCIPAL FINDINGS: A statistically significant improvement in quality of team behaviors was shown between the experimental and control groups following training (p = .012). Subjective workload was not affected by the intervention (p = .668). The clinical error rate significantly decreased from 30.9 percent to 4.4 percent in the experimental group (p = .039). In the experimental group, the ED staffs' attitudes toward teamwork increased (p = .047) and staff assessments of institutional support showed a significant increase (p = .040). CONCLUSION: Our findings point to the effectiveness of formal teamwork training for improving team behaviors, reducing errors, and improving staff attitudes among the ETCC trained hospitals. PMID- 12546287 TI - Specialty health care, treatment patterns, and quality: the impact of a mental health carve-out on care for depression. AB - OBJECTIVES: To assess the effect of a mental health carve-out on treatment patterns and quality of care for outpatient treatment of depression. DATA SOURCES: Outpatient and pharmaceutical claims from September 1993 through March 1997 for one large managed care organization in the Midwest that carved-out mental health and substance abuse benefits in September 1995. RESEARCH DESIGN: Using the treatment episode as the unit of analysis (n = 1,747), changes in treatment patterns associated with the change to a carve-out were evaluated. Logistic regression was used to assess whether in the postperiod a treatment episode was more likely to be treated with (1) an antidepressant and (2) a type and intensity of treatment with proven efficacy. To strengthen confidence in a causal relationship, I search for structural breaks in treatment patterns across a wide range of dates, assuming no a priori knowledge of the timing of the impact of the carve-out. RESULTS: I find the carve-out to be associated with an increase in the use of drug treatments. Although I find a decrease in the use of guideline level treatment over the entire study period, there is an increase in the number of episodes treated with guideline-level treatment over what would be the case in the absence of the carve-out. CONCLUSIONS: The increase in the use of drug treatments suggests previous research that excluded these costs may have overestimated the savings attributable to carve-outs. Guideline-level care appeared to increase as a result of carve-out implementation suggesting the use of management and specialization to reduce costs is not antithetical to quality improvement. PMID- 12546288 TI - An observational study of antibiotic prescribing behavior and the Hawthorne effect. AB - OBJECTIVES: To assess whether prospective, observational study procedures, including questionnaires and audio recording, are associated with different patterns of physician diagnostic decision making and antibiotic prescribing. DATA SOURCES/SETTING: (1) Survey data from a prospective observational study of treatment patterns for children with acute upper respiratory illnesses (10/96 3/97) and (2) retrospective medical record abstraction data of nonobserved encounters for the same problems occurring during (10/96-3/97) and one year after (10/97-3/98) the observational study period. Ten pediatricians in two community practices were studied. STUDY DESIGN: Patterns of diagnoses recorded in the medical record and antibiotics ordered for visits occurring outside of the observational study (same time period and one year later) were compared with the pattern of diagnoses and antibiotics ordered during the observational study. DATA COLLECTION/EXTRACTION METHODS: For the observational study (10/96-2/97), diagnosis and treatment choices were obtained from questionnaires completed by physicians immediately following the visit. For the nonstudy encounters (10/96 3/97 and 10/97-3/98), data were abstracted from medical records one year after the observational study was completed. PRINCIPAL FINDINGS: The proportion of viral cases in which an antibiotic was prescribed was 29 percentage points lower for the observational study compared to the retrospective analysis (p < .05). In one of two study sites, the proportion of cases assigned a bacterial diagnosis was 29 percentage points lower in the observational study period compared to the retrospective study (p <.05). CONCLUSIONS: Observational study procedures including questionnaires and audio recording can affect antibiotic prescribing behavior. Future observational studies aimed at examining the frequency of inappropriate antibiotic prescribing should measure and adjust for the Hawthorne effect; without such adjustments, the results will likely underestimate the true degree of the problem. Future interventions aimed at decreasing inappropriate antibiotic prescribing should consider "harnessing" the Hawthorne effect through performance feedback to participating physicians. PMID- 12546289 TI - Medical expenditures during the last year of life: findings from the 1992-1996 Medicare current beneficiary survey. AB - OBJECTIVE: To compare medical expenditures for the elderly (65 years old) over the last year of life with those for nonterminal years. DATA SOURCE: From the 1992-1996 Medicare Current Beneficiary Survey (MCBS) data from about ten thousand elderly persons each year. STUDY DESIGN: Medical expenditures for the last year of life and nonterminal years by source of payment and type of care were estimated using robust covariance linear model approaches applied to MCBS data. DATA COLLECTION: The MCBS is a panel survey of a complex weighted multilevel random sample of Medicare beneficiaries. A structured questionnaire is administered at four-month intervals to collect all medical costs by payer and service. Medicare costs are validated by claims records. PRINCIPAL FINDINGS: From 1992 to 1996, mean annual medical expenditures (1996 dollars) for persons aged 65 and older were $37,581 during the last year of life versus $7,365 for nonterminal years. Mean total last-year-of-life expenditures did not differ greatly by age at death. However, non-Medicare last-year-of-life expenditures were higher and Medicare last-year-of-life expenditures were lower for those dying at older ages. Last-year-of-life expenses constituted 22 percent of all medical, 26 percent of Medicare, 18 percent of all non-Medicare expenditures, and 25 percent of Medicaid expenditures. CONCLUSIONS: While health services delivered near the end of life will continue to consume large portions of medical dollars, the portion paid by non-Medicare sources will likely rise as the population ages. Policies promoting improved allocation of resources for end-of-life care may not affect non-Medicare expenditures, which disproportionately support chronic and custodial care. PMID- 12546290 TI - Using Medicare data to estimate the prevalence of breast cancer screening in older women: comparison of different methods to identify screening mammograms. AB - OBJECTIVES: To compare different methods for defining screening mammograms with Medicare claims and their impact on estimates of breast cancer screening rates. METHODS: Medicare outpatient facility and physician claims for 61,962 women in 1993 and 59,652 women in 1998 were reviewed for evidence of receipt of screening mammography. We compared the estimates of screening mammography use derived from CPT (Current Procedure Terminology) codes to categorize mammograms as screening or diagnostic versus using an algorithm that uses CPT codes plus breast-related diagnoses in the prior two years. We also compared estimates obtained from review of physician claims alone, facility claims alone, or the combination of the two sources of claims. RESULTS: Use of physician claims alone produced estimates of screening rates similar to rates calculated from use of both physician and outpatient (facility) claims. In 1993, the CPT code for screening mammography underestimated the rate of screening compared to estimates generated by using the algorithm (8.3 percent versus 18.0 percent prevalence, p<0.001). By 1998, the screening prevalence rate generated from using the CPT code for screening mammography more closely approximated the rate generated by the algorithm (23.0 percent versus 25.1 percent). By all methods of estimating screening mammography with Medicare claims, its prevalence increased substantially between 1993 and 1998. CONCLUSION: Providers increased their use of the screening mammography code in their charges to Medicare during the 1990s. This has improved the claims' ability to distinguish screening from diagnostic mammograms, but screening rates computed with claims continue to fall below those generated from self-reports of mammography use among general populations of older women. PMID- 12546291 TI - Measuring what people value: a comparison of "attitude" and "preference" surveys. AB - OBJECTIVE: To compare and contrast methods and findings from two approaches to valuation used in the same survey: measurement of "attitudes" using simple rankings and ratings versus measurement of "preferences" using conjoint analysis. Conjoint analysis, a stated preference method, involves comparing scenarios composed of attribute descriptions by ranking, rating, or choosing scenarios. We explore possible explanations for our findings using focus groups conducted after the quantitative survey. METHODS: A self-administered survey, measuring attitudes and preferences for HIV tests, was conducted at HIV testing sites in San Francisco in 1999-2000 (n = 365, response rate = 96 percent). Attitudes were measured and analyzed using standard approaches. Conjoint analysis scenarios were developed using a fractional factorial design and results analyzed using random effects probit models. We examined how the results using the two approaches were both similar and different. RESULTS: We found that "attitudes" and "preferences" were generally consistent, but there were some important differences. Although rankings based on the attitude and conjoint analysis surveys were similar, closer examination revealed important differences in how respondents valued price and attributes with "halo" effects, variation in how attribute levels were valued, and apparent differences in decision-making processes. CONCLUSIONS: To our knowledge, this is the first study to compare attitude surveys and conjoint analysis surveys and to explore the meaning of the results using post-hoc focus groups. Although the overall findings for attitudes and preferences were similar, the two approaches resulted in some different conclusions. Health researchers should consider the advantages and limitations of both methods when determining how to measure what people value. PMID- 12546292 TI - Measuring preferences for health care interventions using conjoint analysis: an application to HIV testing. AB - OBJECTIVE: To examine preferences for HIV test methods using conjoint analysis, a method used to measure economic preferences (utilities). DATA SOURCES: Self administered surveys at four publicly funded HIV testing locations in San Francisco, California, between November 1999 and February 2000 (n = 365, 96 percent response rate). STUDY DESIGN: We defined six important attributes of HIV tests and their levels (location, price, ease of collection, timeliness/accuracy, privacy/anonymity, and counseling). A fractional factorial design was used to develop scenarios that consisted of combinations of attribute levels. Respondents were asked 11 questions about whether they would choose "Test A or B" based on these scenarios. DATA ANALYSIS: We used random effects probit models to estimate utilities for testing attributes. Since price was included as an attribute, we were able to estimate willingness to pay, which provides a standardized measure for use in economic evaluations. We used extensive analyses to examine the reliability and validity of the results, including analyses of: (1) preference consistency, (2) willingness to trade among attributes, and (3) consistency with theoretical predictions. PRINCIPAL FINDINGS: Respondents most preferred tests that were accurate/timely and private/anonymous, whereas they had relatively lower preferences for in-person counseling. Respondents were willing to pay an additional $35 for immediate, highly accurate results; however, they had a strong disutility for receiving immediate but less accurate results. By using conjoint analysis to analyze new combinations of attributes, we found that respondents would most prefer instant, highly accurate home tests, even though they are not currently available in the U.S. Respondents were willing to pay $39 for a highly accurate, instant home test. CONCLUSIONS: The method of conjoint analysis enabled us to estimate utilities for specific attributes of HIV tests as well as the overall utility obtained from various HIV tests, including tests that are under consideration but not yet available. Conjoint analysis offers an approach that can be useful for measuring and understanding the value of other health care goods, services, and interventions. PMID- 12546293 TI - Gender not a factor for altitude decompression sickness risk. AB - INTRODUCTION: Early, retrospective reports of the incidence of altitude decompression sickness (DCS) during altitude chamber training exposures indicated that women were more susceptible than men. We hypothesized that a controlled, prospective study would show no significant difference. METHODS: We conducted 25 altitude chamber decompression exposure profiles. A total of 291 human subjects, 197 men and 94 women, underwent 961 exposures to simulated altitude for up to 8 h, using zero to 4 h of preoxygenation. Throughout the exposures, subjects breathed 100% oxygen, rested or performed mild or strenuous exercise, and were monitored for precordial venous gas emboli (VGE) and DCS symptoms. RESULTS: No significant differences in DCS incidence were observed between men (49.5%) and women (45.3%). However, VGE occurred at significantly higher rates among men than women under the same exposure conditions, 69.3% and 55.0% respectively. Women using hormonal contraception showed significantly greater susceptibility to DCS than those not using hormonal contraception during the latter two weeks of the menstrual cycle. Significantly higher DCS incidence was observed in the heaviest men, in women with the highest body fat, and in subjects with the highest body mass indices and lowest levels of fitness. CONCLUSION: No differences in altitude DCS incidence were observed between the sexes under our test conditions, although men developed VGE more often than women. Age and height showed no significant influence on DCS incidence, but persons of either sex with higher body mass index and lower physical fitness developed DCS more frequently. PMID- 12546294 TI - Eye tracking, point of gaze, and performance degradation during disorientation. AB - BACKGROUND: The cognitive cockpit concept has been proposed as a potential disorientation countermeasure. It involves monitoring the pilot's physiological, behavioral and subjective responses during disorientation. This data is combined to provide a real time model of pilot state, which is used as a basis for optimizing pilot performance. This study attempts to investigate whether there are consistent behavioral or physiological "markers" that can be monitored during a specific disorientation scenario. METHODS: An Integrated Physiological Trainer with interactive aircraft controls and an eye-tracking device was employed. Fourteen subjects proficient in maintaining straight-and-level flight and who have acquired the skills in changing attitude participated in the study. They were exposed to a flight profile consisting of straight-and-level flying and change in attitude without exposure to a head roll (control condition) and a profile with exposure to a head roll (experimental conditions) during constant yaw rotation. Flight performance parameters and subjects' eye movements and point of gaze behavior were monitored continuously. RESULTS: Immediately on the return to upright head position, all subjects reported a strong apparent pitch displacement that lasted < or = 20 s and a lesser sensation of lateral movement. Significant differences (p < 0.01) were noted on a number of scanning behaviors between the control and the experimental conditions. The appearance of nystagmus was apparent as indicated by the number of involuntary saccades during disorientation. Flight performance decrement in the experimental conditions was reflected by a significant deviation in maintaining airspeed (p < 0.01). CONCLUSION: It appears that the pitch illusion consistently affects visual scanning behavior and is responsible for the decrement in flight performance observed in the simulator. PMID- 12546295 TI - Prophylactic high dose methylprednisolone fails to treat severe decompression sickness in swine. AB - INTRODUCTION: Controlled decompression from saturation conditions is not always an option, particularly in a disabled submarine scenario. Hypothesis Prophylactic high dose methylprednisolone (MP) would improve outcome in severe cases of decompression sickness (DCS). METHODS: Littermate pairs of male Yorkshire swine (n = 86, mean weight +/- SE = 19.3 +/- 0.2 kg) were randomized to one of three groups, then compressed on air to 4.3 ATA (33 msw) for 22 h and brought directly to surface pressure (1 ATA) at 0.9 ATA x min(-1). The MP-50 group received i.v. infusion of 50 mg x kg(-1) of MP dissolved in 60 cc normal saline (NS) immediately prior to the hyperbaric exposure. The NS group received 60 cc NS i.v. immediately prior to the hyperbaric exposure. The MP-10 group received i.v. infusion of 10 mg x kg(-1) MP dissolved in 60 cc NS during the hyperbaric exposure, 7 h before the decompression. RESULTS: Outcomes of severe DCS and death were recorded. NS group: 14 DCS, 4 died; MP-50 group: 19 DCS, 12 died; MP-10 group: 19 DCS, 10 died. Compared with the NS group, logistic regression analysis suggested that animals in the MP-10 group were more likely to get severe DCS and to die (p < 0.01) and animals in the MP-50 group were more likely to die from their disease (p < 0.01). DISCUSSION: Prophylactic high dose MP exerts no protective effect against severe DCS and actually worsens mortality in this model. An earlier group of untreated controls (UC, n = 44, 30 DCS, 11 died, mean weight +/- SE = 19.9 +/- 0.3 kg) exposed to the same profile was also available for analysis. Comparison of the UC and NS animals suggested that pre-dive NS treatment may protect against severe DCS. PMID- 12546296 TI - Color vision with rapid-onset acceleration. AB - INTRODUCTION: Only sporadic information exists concerning perceived color shifts at increased G-loads. The purpose of this study was to investigate whether or not color vision is affected by rapid onset high G7-loads up to +9 Gz, and specifically whether perception of hue changes. METHODS: There were 10 male subjects, 9 with normal color vision and 1 with red-green protanomaly, all accustomed to Gz-loads in a human centrifuge. Each subject was tested on a total of 60 Gz-exposures with 10 s periods at +3, +5, +7, and +9 Gz in the centrifuge on three different days. G-onset rate was 6 G x s(-1). The subjects wore an anti G suit and performed straining maneuvers if necessary to maintain vision. Five square color stimuli of medium saturation (yellow, red, blue, green, and gray) were projected one at a time on a screen in front of the subject, who gave his hue response orally. RESULTS: In 96.6% of exposures to various Gz-loads, the subjects responded by correctly naming colors. (The statistical analyses of the results were done for the subjects with normal color vision, with the protanomalous subject excluded.) Hue shifts occurred at the higher +Gz-levels, including 7.7% of the +9 Gz exposures. Yellow was the hue most frequently perceived as changed. Hue shifts were reported for yellow in 11% and 16% of the +7 and +9 Gz exposures, respectively. Hue shifts at +9 Gz occurred as frequently as blackout and G-LOC together. However, statistical analyses showed no significant effects for +Gz-load. CONCLUSIONS: Absolute identification of the color stimuli of medium saturation was stable and was not significantly affected by the rapid onset +Gz-loads up to and including +9 Gz. PMID- 12546297 TI - Effect of dietary sodium on fluid/electrolyte regulation during bed rest. AB - BACKGROUND: A negative fluid balance during bed rest (BR) is accompanied by decreased plasma volume (PV) which contributes to cardiovascular deconditioning. HYPOTHESIS: We hypothesized that increasing dietary sodium while controlling fluid intake would increase plasma osmolality (POSM), stimulate fluid conserving hormones, and reduce fluid/electrolyte (F/E) losses during BR; conversely, decreasing dietary sodium would decrease POSM, suppress fluid conserving hormones, and increase F/E losses. METHODS: We controlled fluid intake (30 ml x kg(-1) x d(-1)) in 17 men who consumed either a 4.0 +/- 0.06 g x d(-1) (174 mmol x d(-1)) (CONT; n = 6), 1.0 +/- 0.02 g x d(-1) (43 mmol x d(-1)) (LS; n = 6), or 10.0 +/- 0.04 g x d(-1) (430 mmol x d(-1)) (HS; n = 5) sodium diet before, during, and after 21 d of 6 degrees head-down BR. PV, total body water, urine volume and osmolality, POSM, and F/E controlling hormone concentrations were measured. RESULTS: In HS subjects, plasma renin activity (-92%), plasma/urinary aldosterone (-59%; -64%), and PV (-15.0%; 6.0 ml x kg(-1); p < 0.05) decreased while plasma atrial natriuretic peptide (+34%) and urine antidiuretic hormone (+24%) increased during BR (p < 0.05) compared with CONT. In LS, plasma renin activity (+166%), plasma aldosterone (+167%), plasma antidiuretic hormone (+19%), and urinary aldosterone (+335%) increased with no change in PV compared with CONT (p < 0.05). Total body water did not change in any of the subjects. CONCLUSIONS: Contrary to our hypothesis, increasing dietary sodium while controlling fluid intake during BR resulted in a greater loss of PV compared with the CONT subjects. Reducing dietary sodium while controlling fluid intake did not alter the PV response during BR compared with CONT subjects. PMID- 12546298 TI - Probability of survival during accidental immersion in cold water. AB - BACKGROUND: Estimating the probability of survival during accidental immersion in cold water presents formidable challenges for both theoreticians and empirics. A number of theoretical models have been developed assuming that death occurs when the central body temperature, computed using a mathematical model, falls to a certain level. This paper describes a different theoretical approach to estimating the probability of survival. METHOD: The human thermal model developed by Wissler is used to compute the central temperature during immersion in cold water. Simultaneously, a survival probability function is computed by solving a differential equation that defines how the probability of survival decreases with increasing time. The survival equation assumes that the probability of occurrence of a fatal event increases as the victim's central temperature decreases. Generally accepted views of the medical consequences of hypothermia and published reports of various accidents provide information useful for defining a "fatality function" that increases exponentially with decreasing central temperature. RESULTS: The particular function suggested in this paper yields a relationship between immersion time for 10% probability of survival and water temperature that agrees very well with Molnar's empirical observations based on World War II data. DISCUSSION: The method presented in this paper circumvents a serious difficulty with most previous models--that one's ability to survive immersion in cold water is determined almost exclusively by the ability to maintain a high level of shivering metabolism. PMID- 12546299 TI - Incidence of decompression sickness in hypoxia training with and without 30-min O2 prebreathe. AB - BACKGROUND: All naval aviators, navigators, and aircrewmen are required to participate in hypoxia familiarization training. This training is performed in a hypobaric chamber and is considered high risk due to the potential for barotrauma and/or decompression sickness (DCS). Prior analysis of the DCS in U.S. Navy hypobaric chambers revealed a significantly higher incidence among inside observers (IOs) compared with students. In response to these reports, all IOs are required to denitrogenate by breathing 100% oxygen for 30 min prior to altitude exposure (prebreathing). Although the Army, Navy, and Air Force prebreathe for 30 min prior to most hypobaric training exposures, there have been no reports validating the efficacy of this measure. This study examined the incidence of altitude DCS during training exposures to simulated altitudes of 25,000 ft (25k) and 35,000 ft (35k) in IOs and students, some of whom prebreathed and some of whom did not. METHODS: Exposures and DCS cases for a period of 9 yr were tabulated from training reports maintained at the Naval Operational Medicine Institute in Pensacola, FL. Chi-square or Fisher's Exact test was used to compare the data sets and p < or = 0.05 was considered significant. RESULTS: The overall DCS incidence for students and IOs for all chamber profiles was 0.25%. The incidence for 25k was 0.29% for students who did not prebreathe and 0.15% for IOs who did (p = 0.10). Within the student group there was a 0.44% DCS incidence for 25k with no prebreathe and a 0.17% DCS incidence for 35k with prebreathe (p = 0.004). CONCLUSIONS: A 30-min prebreathe prior to altitude exposure appears to contribute to a reduction in the risk of DCS during hypobaric chamber training. PMID- 12546300 TI - In-flight medical events and aircraft diversions: one airline's experience. AB - BACKGROUND: An aging population combined with the increasing mobility of people with acute and chronic illnesses could make an increase in the frequency of in flight medical events aboard commercial aircraft likely. OBJECTIVE: To determine the incidence of each type of in-flight medical complaint, the appropriateness of medical kit contents, which factors lead to aircraft diversion, and which factors effect the appropriateness of the decision to divert. METHOD: Medical complaints reported aboard a sample airline from July 1, 1999 through June 30, 2000 were studied. The frequency of aircraft diversion was related to complaint and medical assistance provided. The appropriateness of the decision to divert was determined as a function of hospital admission rates. RESULTS: There was an incidence of 22.6 medical complaints per million passengers and 0.1 deaths per million passengers. There were 210 diversions per million flights with one of every 12.6 incidents resulting in a diversion. When a passenger volunteer was used, they opened the medical kit 62% of the time. When a physician participated in the decision to divert the hospital admission rate was 49% versus 15% with no physician input. CONCLUSION: Variations in incidence of medical complaints cited in previous studies demonstrate the need for an industry-wide standardized reporting method of in-flight medical events. All in-flight medical complaints could likely have been adequately treated with the contents of the FM's newly mandated medical kits. Physician participation in decisions to divert aircraft should be sought as it is associated with more appropriate divert decisions. PMID- 12546301 TI - Prediction of muscle performance during dynamic repetitive movement. AB - BACKGROUND: During long-duration spaceflight, astronauts experience progressive muscle atrophy and often perform strenuous extravehicular activities. Post flight, there is a lengthy recovery period with an increased risk for injury. Currently, there is a critical need for an enabling tool to optimize muscle performance and to minimize the risk of injury to astronauts while on-orbit and during post-flight recovery. Consequently, these studies were performed to develop a method to address this need. METHODS: Eight test subjects performed a repetitive dynamic exercise to failure at 65% of their upper torso weight using a Lordex spinal machine. Surface electromyography (SEMG) data was collected from the erector spinae back muscle. The SEMG data was evaluated using a 5th order autoregressive (AR) model and linear regression analysis. RESULTS: The best predictor found was an AR parameter, the mean average magnitude of AR poles, with r = 0.75 and p = 0.03. This parameter can predict performance to failure as early as the second repetition of the exercise. CONCLUSION: A method for predicting human muscle performance early during dynamic repetitive exercise was developed. The capability to predict performance to failure has many potential applications to the space program including evaluating countermeasure effectiveness on-orbit, optimizing post-flight recovery, and potential future real-time monitoring capability during extravehicular activity. PMID- 12546302 TI - Lumbar disc volume measured by MRI: effects of bed rest, horizontal exercise, and vertical loading. AB - BACKGROUND: Nutrition to the cells in the disc is partly dependent on fluid flowing out during the day and flowing in during bed rest. In spaceflight there are little or no such diurnal changes, since the gravitational load is essentially zero. HYPOTHESIS: The questions we asked were: 1) How much fluid does the disc gain during the night and how quickly does the disc lose fluid during the following day? 2) Is it possible to carry out, in a reasonable amount of time, an exercise regimen on a spacecraft that would be rigorous enough to expel from the disc the equivalent amount of fluid lost during a normal day's activity? METHODS: in five normal subjects, magnetic resonance imaging was used to measure the volume changes (and the corresponding fluid changes) in the lumbar intervertebral discs (L1/L2, L2/L3, L3/L4, L4/L5) before and after a night's bed rest and again at specific intervals during the course of the day while carrying out three different protocols: walking, carrying a backpack, and exercising in a horizontal position. RESULTS: 1) On average the disc gained 10.6% of its volume (or 0.90 cm3 of fluid) during an overnight bed rest; 2) on rising in the morning and after 8 h (using our walking protocol) the disc volume did not decrease to the volume measured at the end of the previous day; and 3) wearing a backpack that weighed 40% of body weight produced a volume decrease equivalent to the decrease in volume obtained at the end of the day, but it took 4 h to do it. CONCLUSIONS: The ability of the disc to retain water is substantial. It would be difficult to carry out an exercise regimen on a spacecraft that would be rigorous enough to expel from the disc the equivalent amount of fluid lost during a "normal" day's activity on Earth. PMID- 12546303 TI - Civilian use of night vision goggles. AB - Civil aviation operators have expressed an increased interest in conducting night operations with night vision imaging systems. The development of special operational concepts, hardware requirements, training requirements, and regulatory change and oversight is necessary to control for the known performance constraints associated with these devices. In 2001, the Aerospace Medical Association initiated an internal request to review the human factors issues concerning the use of night vision goggles (NVGs) in civilian flight operations. This paper provides some basic information on night vision imaging systems to highlight the Association's position for supporting the appropriate use of NVGs in civilian aviation while concurrently expressing the need for a judicious and studied approach to their deployment. PMID- 12546304 TI - Medications in civil aviation: what is acceptable and what is not? PMID- 12546305 TI - Case from the aerospace medicine residents' teaching file. Pigment dispersion syndrome. AB - A case report is presented of a student naval flight officer with pigment dispersion syndrome. The diagnosis, treatment, and aeromedical disposition of this condition are discussed. PMID- 12546306 TI - You're the flight surgeon. Renal colic. PMID- 12546307 TI - Prevention of carbon monoxide exposure in general and recreational aviation. PMID- 12546308 TI - This month in aerospace medicine history--January 2003. PMID- 12546309 TI - Parachute opening shock simulator to determine cervical injury tolerance. PMID- 12546310 TI - The prevalence and incidence of COPD among urban older persons of Bangkok Metropolis. AB - COPD substantially affects the national healthcare resource and healthcare cost especially among the older persons. Identifying the accurate prevalence and incidence reflects the scale of problem posed by COPD. This epidemiological study using the criteria for diagnosing COPD based on ratio of FEV1.0/FVC less than 70 per cent and the reversibility of less than 15 per cent increase of post bronchodilator FEV1.0 in the absence of parenchymal lesions and cardiomegaly in CXR (PA and lateral view) revealed the prevalence (1998) of COPD among the 3094 older persons aged 60 years and over in the communities of Bangkok Metropolis 10 km around Siriraj Hospital was 7.11 per cent (95% CI: 6.21-8.01), whereas the incidence (1999) of COPD was 3.63 per cent (95% CI: 2.83-4.43). Both the prevalence and the incidence were increased with increasing age. The disease occurred predominantly among male smokers. The distribution of mild : moderate : severe COPD in the prevalence study was 5.6:2.2:1. The current findings also suggest that tobacco smoking is the prime important cause of COPD and the indoor pollution especially cooking smoke is not significant. In particular, the unexpectedly high incidence compared with prevalence in this population probably represents the warning message to the national policy maker for prompt and effective health promotion and disease prevention to prevent further social and economic loss. PMID- 12546312 TI - Efficiency of the glutaraldehyde test strip for monitoring the concentration of glutaraldehyde in reused solutions for disinfecting endoscopes. AB - BACKGROUND: Glutaraldehyde has been widely used for low-temperature disinfection of endoscopes. The current practice at Siriraj Hospital is to change the glutaraldehyde solution every 21 days or when the solution appears turbid. The disadvantages of this practice include inadequate disinfection of endoscopes if the concentration of glutaraldehyde in a reused solution is insufficient or wasted if the discarded solution is still active. OBJECTIVE: To determine the efficiency of a glutaraldehyde test strip (GTS) in monitoring the amount of glutaraldehyde in a reused solution for disinfecting endoscopes. METHOD: Reused glutaraldehyde solutions for disinfecting bronchoscopes, gastroscopes and colonoscopes were tested for the concentration of glutaraldehyde with a GTS thrice weekly for the first week and then every working day up to 56 days. If the GTS indicated a concentration of glutaraldehyde > or = 1.8 per cent after 21 days, 5 ml of the solution was taken to the laboratory to determine its mycobactericidal activity. RESULTS: All samples of the reused glutaraldehyde solution up to 56 days with a concentration of > or = 1.8 per cent glutaraldehyde on GTS from testings showed mycobactericidal activity. If the glutaraldehyde solution was reused for up to 28, 42 or 56 days, it could save 9,603; 22,813 and 29,415 baht per year respectively for the gastroscopy and colonoscopy units. The corresponding figures were -949; 2,726 and 4,564 baht per year for the bronchoscopy unit. It is estimated that up to 400,000 baht per year could be saved by adopting the strategy of GTS monitoring in all endoscopy units at Siriraj Hospital. CONCLUSION: The current strategy of discarding reused glutaraldehyde solution in the gastroscopy, colonoscopy and bronchoscopy units at Siriraj Hospital may be inappropriate since the reused solution is still mycobactericidal for up to 56 days. PMID- 12546311 TI - Comparative study of low-dose oral granisetron plus dexamethasone and high-dose metoclopramide plus dexamethasone in prevention of nausea and vomiting induced by CHOP-therapy in young patients with non-Hodgkin's lymphoma. AB - Standard-dose (2 mg/day) oral granisetron seems to have more antiemetic efficacy than that of high-dose (0.5-1 mg/kg/dose) metoclopramide in moderately emetogenic chemotherapy. However, the cost of oral granisetron is much higher than that of metoclopramide so the authors tried to overcome this disadvantage by dose reduction and adding dexamethasone to enhance the antiemetic effect of oral granisetron. Twenty four young patients (aged < 50 years), with non-Hodgkin's lymphoma receiving CHOP-therapy were enrolled and evaluated in a randomized, double-blind, crossover study comparing the antiemetic efficacy, toxicity and patients' preference of a combination of low-dose oral granisetron plus intravenous dexamethasone (gran/dex) with a combination of high-dose metoclopramide plus intravenous dexamethasone (met/dex) on days 1-5 after chemotherapy. The acute, delayed (day 2-5) and 5-day total control of nausea and vomiting in the gran/dex group were significantly higher than those of the met/dex group (75.0% vs 25.0%; p-value = 0.004, 79.2% vs 33.3%; p-value = 0.007 and 75.0% vs 25.0%; p-value = 0.004, respectively). Except for extrapyramidal reactions in the met/dex group, the side effects in both groups were comparable. The mean total score of antiemetic preference in the gran/dex group was also significantly higher than that of the met/dex group (9.0 vs 7.5; p-value = 0.004). In conclusion, low-dose oral granisetron combined with intravenous dexamethasone had significantly higher protective effects against both acute and delayed nausea and vomiting induced by CHOP-therapy. Thus, this regimen may be considered as an alternative outpatient antiemetic treatment for young patients with non-Hodgkin's lymphoma. PMID- 12546313 TI - Effectiveness of intravenous meperidine for pain relief in the first stage of labour. AB - OBJECTIVE: To determine the effectiveness and side effects of intravenous meperidine in labour pain relief. MATERIAL AND METHOD: A double blind, randomized controlled trial was conducted in 84 parturients, using normal saline as control. Visual analogue scale, postpartum parturients' opinion of effectiveness, sedative scores, nausea/vomiting, dizziness, delivery method, Apgar scores, and naloxone prescription were assessed. RESULT: There were no statistically significant differences between the mean and median of visual analogue scale of meperidine and control group. In addition, the sedative scores, nausea/vomiting and dizziness in the meperidine group occurred more than those in the control group significantly. Even mean of the pain increment in the meperidine group was less than those in the control group (p < 0.05). The parturients' opinion on the effectiveness of pain relief during labor within 24 hours of the first postpartum day was only 23.80 per cent in the meperidine group, however, it was statistically significantly different when compared to 7.10 per cent in the control group. CONCLUSION: Intravenous meperidine exhibited the effectiveness of pain relief of only 23.80 per cent of the subjects, in addition, it may cause many side effects. PMID- 12546314 TI - Double phototherapy in jaundiced term infants with hemolysis. AB - OBJECTIVE: To compare the efficacy of double phototherapy and conventional phototherapy in term newborn infants with hemolytic jaundice. METHOD: Full-term infants with evidence of severe hemolysis on the peripheral blood smear, whose serum bilirubin levels were between 15-21 mg/dl were divided into 2 groups, depending on the availability of the phototherapy bed. Group 1 infants received double phototherapy consisting of conventional (single) phototherapy plus an extra light source from a phototherapy bed (Medela Billibed, Switzerland). Group 2 infants received only conventional phototherapy. Maternal and infants' blood groups and Rh, direct Coomb's test, G6PD screening test and hematocrit were determined on every infant. Phototherapy was given until the serum bilirubin level dropped to < or = 13 mg/dl. Exchange transfusion was indicated when the serum bilirubin level was > or = 21 mg/dl after phototherapy had been given for 4 6 hours. RESULTS: There were 110 infants included in this study, 62 and 48 in group 1 and 2 respectively. There was no statistical difference in terms of birth weight, sex ratio, proportion of breast feeding infants and the initial hematocrit level. However, the initial mean +/- SD of bilirubin level of group 1 infants was higher than that of group 2, (17.7 +/- 1.6 mg/dl vs 16.2 +/- 0.9 mg/dl, p < 0.001). Causes of hemolysis could be determined in 74 infants; 27 (24.5%), 39 (35.5%) and 8 (7.3%) infants had ABO incompatibility, G6PD deficiency and both ABO incompatibility and G6PD deficiency respectively. Rate of bilirubin reduction in group 1 infants was significantly faster, (3.3 +/- 2.4 mg/dl/24 h vs 2.1 +/- 1.1 mg/dl/24 h, p < 0.01). Duration of phototherapy was also shorter in group 1 infants, (45.8 +/- 29.7 hours vs 58.5 +/- 26.0 hours, p < 0.05). Four infants failed to respond to the phototherapy that was assigned. Two infants in group 2 had to be switched to receive double phototherapy because of rapid rising of serum bilirubin. One in each group needed exchange transfusion. CONCLUSION: The study demonstrated that double phototherapy is more efficient than conventional phototherapy in term infants with severe jaundice caused by hemolysis. PMID- 12546315 TI - Modified cox maze procedure for atrial fibrillation with mitral valve diseaset. AB - OBJECTIVE: To determine early term results of the modified Cox maze procedure for curing atrial fibrillation (AF) associated with mitral valve disease. METHOD: Between January and December 2000, 10 consecutive patients with AF underwent the modified Cox maze procedure with mitral valve repair in 2 and replacement in 8. The associated procedure included 3 aortic valve replacements, 1 tricuspid annuloplasty, and 2 atrial septal defect closures. There were 5 males and 5 females, with ages ranging from 19 to 52 years (mean = 38.3 years). Pre-operative existing AF time varied from 4 to 146 months (mean = 41.1 months), and left atrial dimension varied from 4.50 to 6.89 cm (mean 5.51 cm). The authors modified the maze atriotomies to preserve the sinus node artery and used cryoablation, incision and suture to simplify the procedures. RESULTS: Seven cases (70%) regained sinus rhythm and 3 cases (30%) still remained in AF. Two cases (double valve replacement) needed intraaortic balloon pump. Two of them developed cardiac tamponade 1 month post-operatively, (one in sinus rhythm case and another one in AF). Those 3 patients who remained in AF had a longer pre-operative existing AF time and larger left atrial dimension but it was insignificant (p = 0.227 and p = 0.187 respectively). There was no early or late mortality. CONCLUSION: The pertaining results suggest that the modified Cox maze procedure has satisfactory effectiveness to cure atrial fibrillation, restore atrioventricular synchrony and preserve atrial transport function in the patients having AF associated with mitral valve disease. PMID- 12546316 TI - Botulinum A toxin therapy on esotropia in children. AB - A prospective descriptive study was conducted to determine the results of Botulinum Toxin Type A (BTA) injection on esotropia in children, as this may be an alternative to incisional surgery for strabismus. Between September 1998 and February 2002, eleven patients (6 boys and 5 girls) with esotropia were treated with BTA at the Department of Ophthalmology, Songklanagarin Hospital. The average ages at the time of the first and second treatments were 26.8 months (range 14-40 months) and 32.3 months (range 19-54 months), respectively. An open sky procedure was used to inject the BTA. Alignment within +/- 10 prism of orthotropia was considered a successful outcome of BTA treatment in the patients. If the result of the first injection was an incomplete paralysis and the strabismus remained undercorrected, the subsequent dose was increased up to double the initial dose. The mean deviation angle was 40.4 prism diopter (PD) before the first injection and 24.5PD before the second injection. Eight of 11 (72.7%) cases had a sucessful outcome. BTA injection in childhood esotropia may be an alternative to incisional strabismus surgery. PMID- 12546317 TI - Family protective-risk index and its implications. AB - The study aimed to develop an index for differentiating the levels of a family at risk of affecting child development to be applied as a screening tool for primary care workers to identify families that need urgent help. The Family Protective Risk Index (FPRI) was developed from 8 family factors; i.e. mother's education, father's education, family income sufficiency, type of family, family relations, stressful life events in the family, child rearing and physical environment at home that were related to child development in any age group (1-<3 years, 3-<6 years and 6-12 years). Each factor was given a score of 0 or 1 and the scores of FPRI were between 0-8. The family with a lower FPRI score would have a higher risk while the family with a high FPRI score would have more security. The cut off point of FPRI was determined by calculating the sensitivity, specificity, positive predictive value, and negative predictive value. It was later found that the appropriate cut off point for prediction was 6. The 6th FPRI score had a suitable sensitivity to be used for identifying families that need close assistance in order to prevent the slow growth and development of children. PMID- 12546318 TI - Causes of death and poor survival prognostic factors in thai patients with systemic sclerosis. AB - Causes of death and poor prognostic factors for patients with systemic sclerosis (SSc) were studied in 222 cases. Their mean age at the onset and duration of disease was 48.9 +/- 12.0 years and 23.3 +/- 29.3 months, respectively. Fifty three per cent were diffuse subtype. Patients with diffuse SSc had more digital pitting scars and more muscle, heart, lung, and esophageal involvement than those with limited subtypes (p < or = 0.02). One hundred and six patients were lost to follow-up. With a median follow-up duration of 25 months, 31 of the remaining 116 patients (26.7%) died. SSc related death occurred in 18 cases, in which the lung, heart and kidney (renal crisis) were the major causes. Infection contributed to the remaining 13 deaths. When compared with living patients, using a univariate analysis, factors associated with a reduced survival rate were age of > 45 years at the onset, diffuse skin thickness, and lung, gastrointestinal tract, heart, kidney and muscle involvement (p < or = 0.001). In the multivariate analysis, only age of > 45 years at onset and cardiac involvement remained poor prognostic factors (p = 0.04 and 0.001, respectively). PMID- 12546319 TI - The European Organization for Research and Treatment of Cancer quality of life questionnaire: translation and reliability study of the Thai version. AB - In the past decade, increasing attention is being given to more systematic and quantitative ways to evaluate explicitly the impact of disease and medical interventions on quality of life (QOL). Pertaining to the field of oncology, two relatively new instruments--the European Organization for Research and Treatment of Cancer (EORTC) QLQ-C30 and the FACT-G, have received growing attention and appear to be excellent QOL instruments in clinical settings. FACT-G has already been validated and has been used in Thailand. Thus in the present study, the English version of the EORTC quality of life questionnaire (QLQ-C30) was translated into Thai and the initial descriptive statistic and scale reliability were reported. Mean score in this study of 75 cancer patients was comparable with the original report. Cronbach's alpha coefficient for multi-item scales range from 0.64 to 0.89. The validity of this translated version will be reported at a later date. The initial findings of the present study indicate that the Thai version of the EORTC QLQ-C30 is reliable. A validating process of this version is in progress with active patients accrual ongoing at present. PMID- 12546320 TI - Ulcerated hemangiomas: clinical features and management. AB - Hemangioma is the most common tumor of infancy and ulceration is the most frequent complication of hemangioma. The purpose of this report was to review the clinical features and management of ulcerated hemangioma. A retrospective study of ulcerated hemangioma at the outpatient pediatric dermatology clinic, King Chulalongkorn Memorial Hospital from 1992 to 2001 was performed. The medical records of 41 patients were reviewed. Twenty-eight females and 13 males were seen with a female/male ratio of 2.2: 1. Superficial hemangioma was the most common type (51%). Head and neck were the most frequently involved sites, affecting 21 cases (51%). Ulcerated hemangiomas were found on extremities, torso, and perineum of 9 patients (22%), 6 patients (15%) and 5 patients (12%), respectively. Ulceration was developed at a mean age of 3.9 months (range, 1-7 months). Conventional treatment with topical and/or systemic antibiotic was successfully used in 19 patients (46%). Oral prednisolone was used in 4 patients with large hemangiomas. The mean duration of treatment with oral prednisolone was 12 months. Flashlamp-pumped pulsed-dye laser (FPDL) was used in 16 patients who failed topical and/or systemic antibiotic. Fifteen patients (94%) healed within 1 to 3 treatments. Excision was done on a small superficial hemangioma on the cheek. In conclusion, head and neck were the most common sites of ulcerated hemangioma. Topical antibiotics and/or systemic antibiotics could be used as the first-line management of ulcerated hemangioma. FPDL was very effective in the treatment of ulcerated hemangioma. PMID- 12546321 TI - Primary hypomagnesemia in Thai infants: a case report with 7 years follow-up and review of literature. AB - A female Thai baby born to non-consanguineous parents, presented with primary hypomagnesemia at 10 weeks of age, and suffered recurrent convulsions that responded to magnesium supplementation. She was found to have hypomagnesemia (Mg 0.35-1.02 mEq/L) and a low urinary magnesium excretion of less than 10 mg per day, or urinary Mg/Cr that ranged from 0.005-0.01 mg/mg. Intermittent hypomagnesemia and one episode of hypocalcemia with occasional convulsions developed, due to irregular consumption of oral magnesium sulfate, which had a bitter taste, caused frequent loose stools and black staining of the teeth. Better compliance after switching from magnesium sulfate to magnesium oxide resulted in an increased level of serum magnesium and the gradual disappearance of the black staining of the teeth and frequent loose stools. The patient required an oral elemental magnesium dosage of 15-30 mg/kg/day to maintain the serum magnesium level at between 1.02-1.33 mEq/L and keep her free from convulsions. The follow-up period was 7 years during which the patient showed normal physical growth and a mild degree of mental retardation. PMID- 12546322 TI - Bilateral carotid stenting prior to coronary artery bypass graft: a case report. AB - BACKGROUND: Carotid stenosis is an independent possible complication of the central nervous system of patients after receiving a coronary artery bypass graft (CABG). The risk increases when the patient has bilateral carotid stenosis even if asymptomatic. CASE REPORT: A 76 year-old female was admitted because of unstable angina. The coronary angiography showed triple vessel disease and required CABG for revascularization. Her physical examination revealed bilateral carotid bruits. She did not have any history of neurological deficit. Carotid Doppler showed critical stenosis of bilateral carotid arteries. The carotid angiography demonstrated 70 per cent diameter stenosis of both internal carotid arteries just above the bifurcation of the external carotid artery. A 7 x 20 mm self-expandable Smart stent was implanted first in the right carotid artery with good angiographic result. Five days later, another 7 x 20 mm self-expandable Smart stent was implanted in the left carotid artery without residual stenosis. The patient did not have any cardiovascular complications. CABG was performed 2 weeks later with a good result. The patient was discharged 10 days after CABG. CONCLUSION: Bilateral carotid stenting is feasible and produces an acceptable outcome. This procedure is an alternative treatment for preventing stroke during CABG surgery. PMID- 12546323 TI - Bilateral tubal pregnancies after tubal sterilization in a human immunodeficiency virus seropositive woman. AB - Bilateral tubal ectopic pregnancies are rare occurrences. Bilateral tubal ectopic pregnancy in a patient who had undergone tubal sterilization is also very rare. The authors report a case of a 32-year-old, gravida 2, parity 1, human immunodeficiency virus seropositive who had previously undergone tubal sterilization. She presented with abdominal distention and vaginal bleeding after 12 weeks of amenorrhea. Pre-operative diagnosis was ruptured ectopic pregnancy. Emergency exploratory laparotomy and bilateral salpingectomy were performed. The pathology report confirmed bilateral tubal pregnancies. She was well at the fifth day of discharge and four-week follow-up. Ectopic pregnancy should always be suspected in reproductive-age patients presenting with amenorrhea even if they have undergone tubal sterilization. To the authors' knowledge, there has been no report of bilateral tubal pregnancy in a patient with human immunodeficiency virus seropositive after tubal sterilization. PMID- 12546324 TI - Pseudo-Meigs' syndrome secondary to subserous myoma uteri: a case report. AB - A 45 year-old Thai woman, gravida 5, para 5 presented with a huge pelvic mass as well as ascites and right pleural effusion. Right thoracocentesis was performed pre-operatively and malignant cells could not be detected on the cytological examination of the pleural fluid. Surgical exploration revealed a large pedunculated subserous leiomyoma of the uterus without malignant transformation. Total abdominal hysterectomy with bilateral salpingooophorectomy and appendectomy were performed. Both ascites and pleural effusion resolved post-operatively and did not recur during the 12-month follow-up. This case demonstrated the extremely rare case of pseudo-Meigs' syndrome caused by a subserous uterine leiomyoma. PMID- 12546325 TI - Box elder. PMID- 12546326 TI - Does air pollution cause asthma exacerbations in children? PMID- 12546327 TI - Children, their school environment, and asthma. PMID- 12546328 TI - House-dust mite allergen exposure and the development of asthma in children. PMID- 12546329 TI - Acute asthma care interventions in improving asthma outcomes: putting guidelines into action. PMID- 12546330 TI - Optimal management improves asthma morbidity and mortality. PMID- 12546331 TI - A significant evolutionary step in the specialty of allergy and immunology. PMID- 12546332 TI - Mycophenolate mofetil: new applications for this immunosuppressant. AB - OBJECTIVES: This article should familiarize the reader with the pharmacokinetics, mechanisms of action, potential toxicities, and current and future applications of mycophenolate mofetil. DATA SOURCES: The current medical literature. RESULTS: Mycophenolate mofetil is an immunosuppressive agent that has been used for transplant recipients. More recently, this agent has been used to treat several inflammatory conditions. CONCLUSIONS: Mycophenolate mofetil has the potential to be a useful agent in the treatment of several inflammatory conditions, including systemic lupus erythematosus. PMID- 12546333 TI - Montelukast reduces peripheral blood eosinophilia but not tissue eosinophilia or symptoms in a patient with eosinophilic gastroenteritis and esophageal stricture. AB - BACKGROUND: Eosinophilic gastroenteritis (EG) is an uncommon entity of which the pathogenesis is unclear. As no controlled treatment trials exist, treatment of EG remains largely empiric. Limited results have been achieved with oral cromolyn, ketotifen, and other antihistamines. Oral corticosteroids are effective, but long term use is complicated by side effects including growth retardation, diabetes, and osteoporosis. OBJECTIVES: We sought to determine whether treatment with montelukast would improve symptoms and decrease both peripheral blood and tissue eosinophilia (TE) in a patients with steroid-dependent EG for 20 years complicated by esophageal stricture. METHODS: In an unblinded, n = 1 trial, we treated the patient for 5 months with montelukast (20 to 30 mg daily) while his baseline dose of prednisone (10 mg daily) was continued. Complete blood counts and symptoms were monitored weekly. Esophageal biopsies were obtained before and after 5 months of therapy with montelukast. After the posttreatment biopsy was obtained, montelukast was discontinued. Outcome measures included patient symptoms and peripheral and tissue eosinophil counts. RESULTS: During treatment with montelukast, the mean peripheral blood eosinophil count fell from 5,064 cells/microL (average 28 determinations over 20 years; range 1,408 to 12,500 cells/microL) to 1,195 cells/microL (average 14 determinations over 16 weeks; range 556 to 2,193 cells/microL), a 76% reduction. The corresponding TE as calculated from esophageal biopsies was 31 eosinophils/high power field before and 70 eosinophils/high power field after treatment. The patient noted no appreciable improvement in esophageal symptoms. CONCLUSIONS: Montelukast dramatically reduced peripheral blood eosinophilia, but did not affect TE or symptoms in this patient with severe, long-standing EG complicated by esophageal stricture. PMID- 12546334 TI - Relationship of outdoor air quality to pediatric asthma exacerbations. AB - BACKGROUND: Although exposure to outdoor air pollutants has been shown to be associated with exacerbations of asthma, there are relatively few admissions for asthma to Cincinnati Children's Hospital, Cincinnati, OH during the summer months when air quality tends to be worst. OBJECTIVE: The objective of this study was to determine the relationship of outdoor air quality parameters to asthma exacerbations in children. METHODS: The number of emergency room visits and hospitalizations for asthma were determined by review of emergency department logs and the hospital computer database. Outdoor air concentrations of ozone, particulates of < 10 microm diameter (PM-10), pollens, and fungal spores were obtained from the Hamilton County Department of Environmental Services. Multiple regression analysis was performed, looking for relationships between the daily number of asthma visits and the air quality data for the same day and for 1 through 5 days before the visits. RESULTS: A significant association was found between the number of asthma visits and the daily pollen count (P = 0.014, SE = 0.001). The effect was stronger for visits 1, 2, and 3 days after the pollen count (P < 0.001 for pollen count lagged 3 days). High PM-10 counts were synergistic with the pollen count as a predictor of asthma visits. There was no association between asthma visits and the ozone concentration or fungal spore count. CONCLUSIONS: Exacerbations of asthma severe enough to require visits to the hospital were associated with elevated concentrations of airborne pollens and particulates, with a significant delayed effect. Ozone, in the concentrations measured here, was not a risk factor for severe asthma exacerbations in children. PMID- 12546335 TI - Environmental allergens and asthma in urban elementary schools. AB - BACKGROUND: Asthma in school children is rising, and indoor allergens are very common triggers of asthma attacks; however, the risk of the school environment on asthma has not been well studied. OBJECTIVE: To determine the presence and the levels of common aeroallergens in schools, where asthma prevalence rates are high. METHODS: Settled dust samples were collected from 12 Baltimore City public elementary schools, and they were analyzed for the following allergens: cockroaches (Bla g 1/2), dust mites (Der f 1/p 1), dog (Can f 1), cat (Fel d 1), and mouse (Mus m 1). School asthma prevalence rates were correlated with allergen levels, and association between allergen levels and other risk factors present in the schools' environment was examined. RESULTS: The mean and range levels were 1.49 U/g (0 to 8) for Bla g 1/2; 0.38 microg/g (0 to 11.9) for the Der f 1/p 1; 1.44 microg/g (0.1 to 9.6) for Can f 1; 1.66 microg/g (0.2 to 12) for Fel d 1; and 6.24 microg/g (0.3 to 118.3) for Mus m 1. Dust mite, cat and dog allergens were significantly in rooms with carpet and/or area rugs, compared to rooms with bare floors (P < 0.05). Asthma prevalence rates varied from 11.8 to 20.8% between schools and positively correlation with the mean levels of Bla g 1/2 in the schools (P = 0.001). CONCLUSIONS: Common allergens that are known to trigger asthma were detected in all school environments, where asthma prevalence rates were high. However, the overall allergen levels were low, indicating that other factors, including exposures in the homes of asthmatic patients, may have more relevance to sensitization and symptoms than school exposures. PMID- 12546336 TI - Relationship of house-dust mite allergen exposure in children's bedrooms in infancy to bronchial hyperresponsiveness and asthma diagnosis by age 6 to 7. AB - BACKGROUND: Several studies have suggested that exposure to house-dust mite (HDM) allergen in infancy increases the risk of developing asthma. OBJECTIVE: To determine whether exposure to higher levels of dust mite in infants increased the risk of developing bronchial hyperresponsiveness (BHR) or physician-diagnosed asthma by age 6 to 7 years. METHODS: A health maintenance organization-based cohort of 97 middle class suburban children born from 1987 to 1989 with a high cord blood immunoglobulin E, defined as > or = 0.56 IU/mL, was followed as a part of the Childhood Allergy Study. During the first 2 years of life, monthly bedroom dust samples were collected and analyzed for Der f 1 and Der p 1. Between 6 and 7 years of age, 64 of the original cohort answered a questionnaire used to determine the presence of physician-diagnosed asthma, underwent clinical examination, skin prick testing, and methacholine inhalation challenge. Mann Whitney tests were used to compare Der f 1 and Der p 1 levels in homes of children with and without BHR, and those with and without physician-diagnosed asthma. RESULTS: In all, 1,421 dust samples were collected and assayed. No significant differences were seen in either the mean, maximum, or minimum dust mite allergen levels in homes of children with versus without BHR, or children with versus without asthma. However, sensitization to HDM was associated with physician-diagnosed asthma (P < 0.05). CONCLUSIONS: When compared with other studies, we were able to more accurately estimate the level of dust mite allergen exposure through repeated sampling over a relatively long period, incorporating seasonal variations. Although HDM sensitization and asthma were concurrently related, we were unable to find any relationship between level of HDM allergen exposure in children's bedrooms in early childhood and development of BHR or physician-diagnosed asthma by age 6 to 7 years. PMID- 12546337 TI - Factors associated with emergency department use in asthma: acute care interventions improving chronic disease outcomes. AB - OBJECTIVE: To provide a literature review of the factors associated with childhood asthma-related emergency department (ED) visits and to identify elements of effective ED interventions that reduce the frequency of childhood ED visits while increasing primary health care utilization. DATA SOURCE: English Medline articles from 1990 that cross-referenced with the terms asthma, emergency, intervention, pediatric, and/or acute care. Experts in the field of allergy and asthma were also consulted. STUDY SELECTION: Childhood asthma interventions in the ED. RESULTS: Factors associated with childhood asthma related ED visits include being impoverished, being exposed to allergens, receiving Medicaid or lacking insurance, being noncompliant with self-management skills, and having an African-American heritage. Other minorities may also be at risk, but further investigation is required to determine the extent. Attempts to link the patient to primary health care by the ED staff resulted in increased adherence to followup care. CONCLUSIONS: The ED provides an opportunity to help patients and families deal with asthma to improve their quality of life. Further, current studies demonstrate that the ED is an appropriate setting for an intervention that links the patient back to the primary health care provider. More research is needed on the appropriate educational messages to be delivered in ED. Also, barriers to followup care and regular use of a primary health care provider need to be identified so that future intervention designs can address these issues. PMID- 12546338 TI - Trends and ethnic differences in asthma hospitalization rates in Singapore, 1991 to 1998. AB - BACKGROUND: A few reports have indicated that asthma hospitalization rates in several countries have stopped rising or started falling in the 1990s. AIM: To describe recent trends and ethnic differences in asthma hospitalization rates in Singapore from 1991 to 1998. METHODS: Asthma hospitalization rates in all hospitals were analyzed by age groups, sex, ethnicity, and individual years, using aggregated data for asthma (ICD-9 493 and ICD-10 J45, J46) from 1991 to 1998, when nationwide data from the Central Claims Processing System were available. RESULTS: Between 1991 and 1998 there were a total of 37,615 hospital admissions for asthma, giving an annual average rate of 17.1 hospital admissions per 10,000 persons. Overall, the rates of asthma hospitalization fell by 28% from 21.7 per 10,000 in 1991 to 15.4 per 10,000 in 1998 (3.5% annually). The trends were broadly based across all age, sex, and ethnic groups. Hospitalizations were more common in boys than in girls aged 0 to 4 (male/female ratio 1.69), but less common in men than women aged 35 to 64 (male/female ratio 0.81). Rates of asthma hospital admissions were higher in Malays (32.8 per 10,000) and Indians (40.8 per 10,000) than Chinese (11.9 per 10,000). CONCLUSIONS: In line with findings from several countries, there have been recent declines in rates of hospital admissions for asthma in Singapore in the 1990s. The declines were broadly based across all population subgroups and parallel previously observed declines in mortality in adults. However, considerable ethnic differences in levels of asthma hospitalization still exist. PMID- 12546339 TI - The effects of intranasal triamcinolone acetonide and intranasal fluticasone propionate on short-term bone growth and HPA axis in children with allergic rhinitis. AB - OBJECTIVE: The purpose of this study was to evaluate the effects of triamcinolone acetonide (TAA) and fluticasone propionate (FP) aqueous nasal sprays on short term lower-leg growth velocity and hypothalamic-pituitary-adrenal (HPA-axis function in pediatric subjects. METHODS: In this controlled, double-blinded (TAA) or single-blinded (FP), four-way crossover trial, 59 subjects (mean age: 7.2 years) were randomized to receive each of four 2-week treatments in random order: TAA nasal spray 110 microg, TAA nasal spray 220 microg, FP nasal spray 200 microg, and placebo, administered by a third party once daily with a 2-week washout period between treatments. Lower-leg growth velocity was measured by knemometry, and HPA-axis function was measured using 12-hour overnight urinary cortisol levels. RESULTS: Forty-nine subjects completed all four treatments and were included in the analyses. Mean growth velocity (+/- standard error) was 0.46 (+/- 0.06) mm/week for placebo, 0.37 (+/- 0.06) and 0.31 (+/- 0.06) mm/week for TAA nasal spray 110 and 220 microg, respectively, and 0.37 (+/- 0.06) mm/week for FP nasal spray. The treatment effect on mean growth velocity compared with placebo was -19.6% with TAA 110 microg, -32.6% with TAA 220 microg, and -21.7% with FP; none of these effects was considered statistically or clinically significant according to predefined criteria. No significant differences in changes in urine cortisol/creatinine ratios were observed between TAA 110 microg or 220 microg and placebo (4.38, 3.60, and -0.67, respectively, P > or = 0.157). In contrast, the change in mean urine cortisol/creatinine ratio values for FP ( 3.59) were significantly lower compared with TAA 220 microg (P = 0.033) and placebo (P = 0.003). Knemometry exhibited less time-dependent variability than overnight urinary cortisol measurements. CONCLUSIONS: Neither TAA nor FP had a clinically significant effect on lower-leg growth velocity. In contrast to FP, TAA nasal spray did not significantly affect HPA-axis function when used over a 2 week interval. PMID- 12546340 TI - Geographical variations of asthma and asthma symptoms among schoolchildren aged 5 to 8 years and 12 to 15 years in Palestine: the International Study of Asthma and Allergies in Childhood (ISAAC). AB - BACKGROUND: Many studies demonstrated the existence of geographic differences, within and between countries, in the prevalence of asthma, rhinitis, and eczema. However, in Palestine, there are no comprehensive Palestinian data to compare with those from other regional and international centers. OBJECTIVE: To describe the prevalence of asthma and asthma symptoms in schoolchildren in two districts (Ramallah and North Gaza) in Palestine. METHODS: After a two-stage stratified systematic sampling, approximately 14,500 schoolchildren, from the first and second grades of elementary school (ages 5 to 8 years) and eighth and ninth school grades (ages 12 to 15 years), were invited to participate in a survey using International Study of Asthma and Allergies in Childhood phase III questionnaires and protocols. RESULTS: In general, younger children were reported to have a higher 12-month wheezing prevalence rate than older children (9.6 and 7.2%, respectively), and more physician-diagnosed asthma (8.4 and 5.9%, respectively). However, nocturnal cough and exercise-related wheezing were higher in the older age group compared with younger children. Younger children living in North Gaza district showed slightly higher prevalence rates for asthma and asthma symptoms, but older children had higher rates in Ramallah district. After adjustment using logistic regression analysis, male sex, living in inland areas, and younger age were shown to predict 12-month wheezing and physician-diagnosed asthma. CONCLUSIONS: Palestinian children have asthma symptoms rates that are similar to several countries in the Mediterranean region such as Spain and Turkey, but still lower than other Middle East countries such as Saudi Arabia and Israel. PMID- 12546341 TI - Increased matrix metalloproteinase-9 with elastolysis in nocturnal asthma. AB - BACKGROUND: Matrix metalloproteinase-9 (MMP-9) is capable of degrading elastin, whereas tissue inhibitor of metalloproteinase-1 (TIMP-1) can inhibit MMP-9 activity. We observed reduced airway tissue elastin volume density in six subjects with nocturnal asthma (NA) as compared with seven subjects with nonnocturnal asthma (NNA) and seven normal controls (NL) when endobronchial biopsies were evaluated morphometrically at 4:00 PM and 4:00 AM. OBJECTIVE: We hypothesized that increased metalloproteinases and decreased tissue inhibitors of metalloproteinases in the airways of subjects with NA may be responsible for reduced elastin volume density. METHODS: Ten additional subjects with NA, 10 subjects with NNA, and 7 normal control subjects underwent bronchoscopy with bronchoalveolar lavage at 4:00 PM and 4:00 AM. Levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 were determined in bronchoalveolar lavage by enzyme-linked immunosorbent assay. RESULTS: There was a fourfold circadian increase in bronchoalveolar lavage levels of MMP-9, and there was a twofold increase in MMP 9:TIMP-1 ratio in NA subjects from 4:00 PM to 4:00 AM. There were no circadian changes in the NNA and NL subjects. At 4:00 AM, MMP-9 levels and the MMP-9:TIMP-1 ratio were highest in NA subjects. At 4:00 PM, no significant group differences were observed. The MMP-9 levels positively correlated with the overnight fall in forced expiratory volume in 1 second and the MMP-9/TIMP-1 ratio negatively correlated with the 4:00 AM % predicted forced expiratory volume in 1 second. CONCLUSIONS: Our results from these two pilot studies suggest that increased MMP 9 and decreased TIMP-1 at night in NA may lead to reduced elastin density. PMID- 12546342 TI - Efficacy and safety of clemastine-pseudoephedrine-acetaminophen versus pseudoephedrine-acetaminophen in the treatment of seasonal allergic rhinitis in a 1-day, placebo-controlled park study. AB - BACKGROUND: Allergic rhinitis afflicts more than 40 million people in the United States and is a leading cause of reduced productivity at work and in school. Patients with allergic rhinitis have a wide range of symptoms that are often treated with oral combination products that contain antihistamines, decongestants, and analgesics. OBJECTIVE: To evaluate the onset of action and the extent of efficacy and safety of a combination (CPA) of clemastine (0.68 mg), pseudoephedrine (60 mg), and acetaminophen (1,000 mg) versus a combination (PA) of pseudoephedrine and acetaminophen versus placebo in the treatment of seasonal allergic rhinitis (SAR). The primary goal was to evaluate the benefit of adding clemastine to the PA combination product to treat the symptoms of SAR. METHOD: A 1-day, multicenter, double-blind, double-dummy, randomized, parallel-group park study was organized, and medication was given at 9:00 AM and 3:00 PM. RESULTS: A total of 298 subjects participated at two outdoor facilities. The primary efficacy outcome was the major symptom complex score averaged over the period of 2 to 5 hours after each dose. Mean absolute and percentage reduction in major symptom complex averaged over the period of 2 to 5 hours in the CPA group was significantly superior to those of either the PA (P < 0.01) or placebo (P < 0.03) groups. Somnolence, fatigue, and nausea were the most common volunteered adverse events; only somnolence was significantly greater after CPA than after either PA or placebo. CONCLUSIONS: Treatment with CPA was safe and highly effective in reducing symptoms associated with SAR. It was more effective than either PA or placebo over most of the postdose observation period. PMID- 12546343 TI - Successful desensitization to high-dose methotrexate after systemic anaphylaxis. AB - BACKGROUND: Methotrexate (MTX) is a folic acid antagonist used in high doses as adjuvant chemotherapy for osteosarcoma. It has many known toxicities and has been reported to cause anaphylaxis. We report the first successful desensitization protocol to high-dose MTX. CASE PRESENTATION: The patient is a 22-year-old male with the evaluation of osteosarcoma, T1Gr2NxM0. He received neoadjuvant chemotherapy and later underwent a right above the knee amputation without complication. He was then scheduled to receive adjuvant chemotherapy with high dose MTX, 12 g/m2 body surface area, followed by leucovorin rescue and ifosfamide. He had an immediate hypersensitivity reaction during the initiation of the MTX infusion with diffuse urticaria, facial swelling, cough, and chest tightness. The infusion was terminated and his symptoms abated. He was later skin tested to confirm allergy to MTX. In order for him to receive the needed chemotherapy, we developed a desensitization protocol that was administered in the intensive care unit. Before the infusion he was pretreated with ranitidine, cetirizine, hydroxyzine, montelukast, and prednisone. He underwent the desensitization without incident on repeated infusions. Serum tryptase levels drawn during the infusions were low, signifying controlled mast cell degranulation. CONCLUSIONS: This case describes the original development of a desensitization protocol to high-dose MTX. The successful development and implementation of this protocol will have impact on patients who have anaphylactic reactions to MTX but require this medication for specific diseases. For patients who suffer from osteogenic sarcoma and have anaphylactic reactions to MTX, this desensitization protocol will allow these patients to continue with needed therapeutic or palliative chemotherapy. PMID- 12546344 TI - Spetzler-Martin grades IV and V arteriovenous malformations. PMID- 12546345 TI - Intention-to-treat analysis of Spetzler-Martin grades IV and V arteriovenous malformations: natural history and treatment paradigm. AB - OBJECT: In this study the authors quantified a subgroup of patients with Spetzler Martin Grades IV and V arteriovenous malformations (AVMs) recommended for complete, partial, or no treatment, and calculated the retrospective hemorrhage rate for these lesions. METHODS: Between July 1997 and May 2000, 73 consecutive patients with Grades IV and V AVMs were evaluated prospectively by the cerebrovascular team at Barrow Neurological Institute. Treatment recommendations given to the patients or referring physicians were classified as complete treatment, partial treatment, and no treatment. Retrospectively, the hemorrhage rates associated with these treatment groups were also calculated. In the prospective portion of the study (the intention-to-treat analysis), no treatment of the AVM, was recommended for 55 patients (75%) and partial treatment was recommended for seven patients (10%). Aneurysms associated with an AVM were obliterated by surgical or endovascular treatment in seven patients (10%), and complete surgical removal was recommended for four patients (5%). The overall hemorrhage rate for Grades IV and V AVMs was 1.5% per year. The annual risk of hemorrhage was 10.4% among patients who previously had received incomplete treatment, compared with patients without previous treatment. CONCLUSIONS: The hemorrhage risk of 1.5% per year, which was associated with Grades IV and V AVMs appears to be lower than that reported for Grades I through III AVMs. The authors recommend that no treatment be given for most Grades IV and V AVMs. No evidence indicates that partial treatment of an AVM reduces a patient's risk of hemorrhage. In fact, partial treatment may worsen the natural history of an AVM. The authors do not support palliative treatment of AVMs, except in the specific circumstances of arterial or intranidal aneurysms or progressive neurological deficits related to vascular steal. Complete treatment is warranted for patients with progressive neurological deficits caused by hemorrhage of the AVM. This selection process plays a significant role in the relatively low combined morbidity and mortality rates for Grade IV and Grade V AVMs (17 and 22%, respectively) reported by the cerebrovascular group in both retrospective and prospective studies. PMID- 12546346 TI - The impact of methicillin-resistant Staphylococcus aureus in a neurosurgical unit: a growing problem. AB - OBJECT: Methicillin-resistant Staphylococcus aureus (MRSA) infection is a growing problem worldwide. To investigate the severity of the problem, the authors surveyed the incidence of MRSA colonization and infection in the neurosurgical unit at their institution. METHODS: Patients colonized or infected with MRSA who had been treated in the neurosurgical unit between 1993 and 1999 were retrospectively identified from laboratory records. There were 203 patients with MRSA-positive cultures, and the incidence of infection increased between 1993 (16 cases; 1.9% of admissions) and 1999 (60 cases; 6.7% of admissions). The mean duration of hospital stay was longer in patients with MRSA than in all patients treated in the unit (33.6 compared with 10.3 days, p < 0.001). Methicillin resistant S. aureus was isolated from the nose in 89 patients, the throat in 79, the perineum in 52, surgical wounds in 16, sputum in 15, blood in 10, and from multiple sites in 69 patients. Fifty-six patients (28%) were infected with MRSA, and there were 15 deaths, of which three (20%) were likely to be due to the infection. The sources of MRSA included the neurosurgical ward in 84 patients, the intensive care unit in 28, other hospitals in 39, and the community in 17. The common strains of MRSA isolated were epidemic (E)MRSA- 16 (110 cases) and EMRSA-15 (31 cases). The microorganism was eradicated in 16 cases, not eradicated in 20, and 167 patients were discharged from the hospital before eradication was achieved. All MRSA isolates were sensitive to vancomycin and teicoplanin and there was reduced sensitivity to mupirocin. CONCLUSIONS: Infection with MRSA is a growing problem in the neurosurgical population, and most cases are hospital acquired and are associated with longer hospital stays. Asymptomatic colonization by this organism is far more common than infection of the surgical wound, although there is still morbidity due to MRSA sepsis. Most patients with MRSA are discharged before eradication of infection is achieved, thus increasing the risk that the infection will spread in the community. Strict adherence to the basic principles of infection control is the key to eradication of MRSA. PMID- 12546347 TI - Prognostic value of magnetic resonance imaging-guided stereotactic biopsy in the evaluation of recurrent malignant astrocytoma compared with a lesion due to radiation effect. AB - OBJECT: The prognostic value of differentiating between recurrent malignant glioma and a lesion due to radiation effect by performing stereotactic biopsy has not been assessed. Thus, this study was undertaken to determine such value. METHODS: Between 1995 and 2001, 114 patients underwent magnetic resonance (MR) imaging-guided stereotactic biopsy to differentiate lesions caused by a recurrence of malignant astrocytoma and by radiation effect. All patients had previously undergone tumor resection (World Health Organization Grade III or IV) followed by radiotherapy. Disease diagnosis based on biopsy and patient characteristics were assessed as predictors of survival according to results of a multivariate Cox regression analysis. The diagnosis determined with the aid of biopsy was compared with that established during a subsequent resection in 26 patients. Survival following stereotactic biopsy was markedly increased in patients suffering from radiation effect compared with those harboring recurrent malignant glioma (p < 0.0001). In patients with radiation effect on biopsy, an increasing patient age (p < 0.05), having had two compared with one prior resection (p < 0.05), and a decreasing time from radiotherapy to biopsy (p < 0.001) were factors associated with decreased survival. Nevertheless, in patients with biopsy-defined radiation effect at second progression or with an age younger than 50 years the survival rate remained higher than that in patients with recurrent tumor on biopsy (p < 0.01). A biopsy-based diagnosis of radiation effect obtained less than 5 months after radiotherapy was not associated with an increased rate of patient survival compared with a diagnosis of recurrent malignant glioma on biopsy (p = 0.286). Eighty-six percent of lesions initially determined to be due to radiation effect on biopsy fewer than 5 months after radiotherapy were characterized as recurrent glioma by a mean of 11 months later. In contrast, only 25% of lesions initially diagnosed as attributable to radiation effect on biopsy more than 5 months after radiotherapy were classified as recurrent glioma a mean of 12 months later (p < 0.05). CONCLUSIONS: With the aid of stereotactic biopsy the authors demonstrated prognostic significance in differentiating recurrent malignant astrocytoma from a lesion due to radiation effect in patients presenting more than 5 months after having undergone radiotherapy. In patients who presented earlier than 5 months after radiotherapy, radiation effect on biopsy was not associated with an improved rate of survival compared with that in patients harboring recurrent malignant astrocytoma. PMID- 12546348 TI - Preoperative and postoperative cognitive functioning in patients with frontal meningiomas. AB - OBJECT: There is presently no specific information available concerning the nature and course of cognitive deficits caused by intracranial meningiomas. In this prospective study the authors examined the cognitive functioning of patients with frontal meningiomas. METHODS: Fifty-four patients with frontal meningiomas were examined neuropsychologically before and after neurosurgery. The test battery consisted of standardized instruments including those assessing memory, attention, visuoconstructive abilities, and executive functions. The time period between pre-and postoperative assessment ranged from 4 to 9 months. The patients' performance was compared with the results in 54 healthy adults who were also assessed twice by using the same test battery in a period ranging from 4 to 9 months. In addition, the effect on cognition of meningioma lateralization, localization, lesion size, edema, brain compression, time course, and the occurrence of preoperative seizures was analyzed. CONCLUSIONS: Except in the case of working memory, comparisons of pre- and postoperative assessments of cognition revealed no differences in memory, visuoconstructive abilities, or executive functions, although a postoperative improvement in attentional functions was observed. The results of this study indicate that the surgical removal of frontal meningiomas does not impair patients' cognitive functioning. Furthermore, improvements in attentional functions may occur in these patients. PMID- 12546349 TI - Sex-related differences in patients with severe head injury: greater susceptibility to brain swelling in female patients 50 years of age and younger. AB - OBJECT: The goal of this study was to study the influence of sex and age on factors affecting patient outcome in severe head injury. METHODS: Data from the prospectively conducted international trial of tirilazad mesylate in patients with head injury were analyzed retrospectively. Included were 957 patients, 23% of whom were female and all of whom were between the ages of 15 and 79 years. All patients presented with Glasgow Coma Scale (GCS) scores between 3 and 8 and evidence of structural brain damage and/or subarachnoid hemorrhage (SAH) on the initial CT scan. Frequencies of recognized risk factors, including brain swelling, intracranial hypertension, systemic hypotension, advanced age, SAH, and injury severity (based on GCS scores), as well as dichotomized Glasgow Outcome Scale (GOS) scores (good recovery or moderate disability compared with severe disability, persistent vegetative state, or death) obtained 6 months postinjury were compared between male and female patients. CONCLUSIONS: Overall significantly greater frequencies of brain swelling and intracranial hypertension were found in female compared with male patients (35% compared with 24% [p < 0.0008] and 39 compared with 31% [p < 0.03], respectively). The highest rates were found in female patients younger than 51 years old (38% compared with 24% [p < 0.002] and 40% compared with 30% [p < 0.02], respectively, in male patients younger than 51 years of age). This effect was independent of injury severity (GCS) scores, which were not different in male and female patients. Female patients younger than 50 years tended to have worse outcomes, but the difference was not statistically significant. Thus, female patients who sustain severe head injury, especially (presumably) premenopausal ones aged 50 years and younger, are significantly more likely to experience brain swelling and intracranial hypertension than male patients with a comparable injury severity, suggesting that younger women may benefit from more aggressive monitoring and treatment of intracranial hypertension. PMID- 12546350 TI - Traumatic subarachnoid hemorrhage on the computerized tomography scan obtained at admission: a multicenter assessment of the accuracy of diagnosis and the potential impact on patient outcome. AB - OBJECT: The goal of this study was fourfold: 1) to determine the incidence of traumatic subarachnoid hemorrhage (tSAH) in patients with traumatic brain injury (TBI); 2) to verify agreement in the diagnosis of tSAH in a multicenter study; 3) to assess the incidence of tSAH on the outcome of the patient; and 4) to establish whether tSAH itself leads to an unfavorable outcome or whether it is a sign of major brain trauma associated with severe posttraumatic lesions. METHODS: Computerized tomography (CT) scans obtained in 169 head-injured patients on admission to 12 Italian intensive care units during a 3-month period were examined. The scans were collected for neuroradiological review and were used for the analysis together with data from a multicenter database (Neurolink). A review committee found a high incidence of tSAH (61%) in patients with TBI and a moderate agreement among centers (K = 0.57). Significant associations were observed between the presence and grading of tSAH and patient outcomes, and between the presence of tSAH and the severity of the CT findings. Logistic regression analysis showed that the presence of tSAH and its grading alone do not assume statistical significance in the prediction of unfavorable outcome. CONCLUSIONS: Traumatic SAH frequently occurs in patients with TBI, but it is difficult to detect and grade. Traumatic SAH is associated with more severe CT findings and a worse patient outcome. PMID- 12546351 TI - Aneurysmal subarachnoid hemorrhage in young adults. AB - OBJECT: The authors reviewed the management protocols for young adults who presented with subarachnoid hemorrhage (SAH) at the Regional Neurosurgery Unit in Newcastle during a study period of 9 years. Aneurysmal SAH is uncommon in the age group selected (18-39 years) and, therefore, the performance of these patients has not been extensively reported in the literature. The authors also evaluated the good-grade rebleed rate (an index of management efficiency) in this cohort of patients. METHODS: The Newcastle neurosurgical unit serves a population of close to 3 million people, and an average of 180 patients with SAH are seen each year. The majority of patients are transferred from other hospitals in the region. This study includes patients admitted between January 1990 and December 1998. A total of 1,609 patients were admitted during this period, of whom 295 (18.4%) between the ages of 18 and 39 years constituted the study population of young adults. Two hundred ninety-five young adults presented with SAH; 181 (61.4%) were women and 114 (38.6%) were men, a ratio of 3:2. Of 246 patients in whom this value was recorded, 15 (6.1%) presented with a history of hypertension, and there was an association between hypertension and the occurrence of multiple aneurysms (Fisher two-tailed exact test, p = 0.008). Thirty-five patients (11.9%) presented with a hematoma on computerized tomography scans; of these, 20 (57%) were women and 15 were men. In six patients the lesion had rebled before treatment. The good-grade rebleed rate was three (1.7%) of 178. The overall favorable outcome rate was 83.8% (Glasgow Outcome Scale [GOS] 4 and 5) and unfavorable outcome occurred in 16.2% (GOS 1-3), with a total of 40 deaths in this group (13%). Age had no influence on outcome in young adults. Comparing the outcome at discharge with the follow-up evaluation at 6 months revealed that patients in the moderate and severe disability groups continued to improve and many achieved good recovery. CONCLUSIONS: In this report the authors detail the outcome of a large number of young adults with SAH. The incidence of SAH was higher in the female population, although the ratio was not as high as previously reported. The authors have also demonstrated a progressive increase in the incidence of aneurysmal SAH with age, even in young adults. Hypertension but not age influenced the occurrence of multiple aneurysms. The good-grade rebleed rate is low, although it is not zero. Generally, a satisfactory outcome was obtained and significant continuing improvements were noted between discharge and follow-up evaluation. This reflects the power of recovery in young adults. These are people whose economic productivity and fertility are at peak levels and therefore the financial and social burden occasioned by less-than-perfect outcomes is large. PMID- 12546352 TI - Natural hypothermia immediately after transient global cerebral ischemia induced by spontaneous subarachnoid hemorrhage. AB - OBJECT: Spontaneous subarachnoid hemorrhage (SAH) has an aspect of graded transient global cerebral ischemia. The purpose of the present study was the documentation of sequential changes in body temperature immediately after SAH induced transient global cerebral ischemia in humans. METHODS: Patients admitted within 12 hours after the initial onset of SAH were examined retrospectively (426 patients). Patients with unruptured cerebral aneurysms served as a control group (73 patients). Body temperature measured at the axilla on admission was analyzed. The grade of SAH was established according to the Glasgow Coma Scale (GCS): Grade I, GCS Score 15; Grade II, GCS Score 11 to 14; Grade III, GCS Score 8 to 10; Grade IV, GCS Score 4 to 7; and Grade V, GCS Score 3. The mean body temperature of patients in the control group was 36.49 +/- 0.45 degrees C (mean +/- standard deviation). The mean body temperature of patients in the SAH group who had been admitted within 4 hours of onset for Grades I to V were significantly different (p < 0.001, analysis of variance [ANOVA]): 36.26 +/- 0.7 degrees C, 59 patients; 35.98 +/-0.85 degrees C, 73 patients; 35.52 +/- 0.79 degrees C, 25 patients; 35.9 +/- 1.09 degrees C, 108 patients; and 35.56 +/- 1.14 degrees C, 73 patients, respectively. These values were significantly lower than those in control volunteers, except for patients with Grade I SAH. The reduction in body temperature was unrelated to the location of the cerebral aneurysm and was not the product of circadian rhythm. The temperatures of patients in the SAH group who were admitted beyond 4 hours after onset for each grade were significantly different (p < 0.01, ANOVA): 36.8 +/- 0.91 degrees C, 36 patients; 36.74 +/- 0.68 degrees C, 31 patients; 36.73 +/- 0.38 degrees C, three patients; 37.41 +/- 1.37 degrees C, 17 patients; and 38.9 degrees C, one patient, respectively. These values were significantly higher than those in patients admitted within 4 hours of SAH onset for all grades except Grade V, and significantly higher than control values in patients with Grades I and IV SAH. CONCLUSIONS: These results indicate that body temperature falls and then rises immediately after the SAH-induced transient global cerebral ischemia without cardiac arrest in humans. The reduction in temperature may be a natural cerebral protection mechanism that is activated shortly after ischemic insult. PMID- 12546353 TI - The impact of stereotactic pallidal surgery on the dopamine D2 receptor in Parkinson disease: a positron emission tomography study. AB - OBJECT: The aim of this study was to estimate the impact of stereotactic pallidal surgery on the binding potential of dopamine D2 receptors in patients with advanced Parkinson disease (PD). METHODS: Six patients with advanced PD (three men and three women; mean age 56.7 +/- 9.8 years, Hoehn and Yahr stage 3.3 +/- 1.1/3.9 +/- 1.2 [on/off scores], mean +/- standard deviation) underwent stereotactic pallidal surgery. One underwent right posteroventral pallidotomy (PVP), one received left PVP, three were treated with deep brain stimulation (DBS) of the left globus pallidus internus (GPi), and one with bilateral DBS of the GPi. The binding potential of the dopamine D2 receptors of these patients was determined before and after surgery by using positron emission tomography scanning with 11C-nemonapride and it was compared with the value in eight healthy volunteers. The authors also examined whether changes in the D2 receptor binding potential were correlated with the clinical outcome. The clinical symptoms, especially those in the off state, were significantly improved after surgery. Preoperatively, the D2 receptor binding potential in the putamen was elevated by 27% (p < 0.01) and that in the thalamus was 29% lower than that in controls (p < 0.01). The D2 receptor binding potential in the putamen and thalamus returned to control levels after surgery. The preoperative level of the D2 receptor binding potential in the anterior cingulate cortex was comparable to that of controls, but it declined significantly after surgery, whereas the D2 receptor binding potential in other regions of both hemispheres showed no significant changes after surgery. Although the D2 receptor binding potential did not correlate with the Hoehn and Yahr stage, the Schwab and England score, or the Unified PD Rating Scale (UPDRS) score, a positive correlation was seen between the percent improvement rate of the total UPDRS score in the off state and the percentage change of the D2 receptor binding potential in the putamen (r = 0.773, p = 0.0417 according to the Pearson linear correlation). CONCLUSIONS: The altered dopamine D2 receptor binding potential in the putamen might play a crucial role in clinical improvement after PVP or DBS of the GPi in advanced PD. PMID- 12546354 TI - Cortical reorganization in patients with subcortical hemiparesis: neural mechanisms of functional recovery and prognostic implication. AB - OBJECT: A systematic investigation on cortical reorganization in patients with hemiparesis of a subcortical origin, with special emphasis on functional correlates, was conducted using functional magnetic resonance (fMR) imaging performed on a 3-tesla system specifically optimized for fMR imaging investigation. METHODS: The study group included 46 patients with hemiparesis (25 with right and 21 with left hemiparesis) and 30 age-matched healthy volunteers as controls. All study participants were originally right handed. The characteristics of the lesion were putaminal hemorrhage in 19 patients, thalamic hemorrhage in 10 patients, and striatocapsular bland infarction in 17 patients. Functional recovery in subcortical hemiparesis showed two distinct phases of the recovery process involving entirely different neural mechanisms. Phase I is characterized by the process of recovery and/or reorganization of the primary system. Successful recovery of this system is typically reached within 1 month after stroke onset. Its clinical correlate is a rapid recovery course and significant recovery of function within 1 month of stroke onset. Failure of recovery of the primary system shifts the recovery process to Phase II, during which reorganization involving the ipsilateral pathway takes place. The clinical correlate of Phase II is a slow recovery course with variable functional outcome. CONCLUSIONS: Effective functional organization of the ipsilateral pathway, as identified by linked activation of the ipsilateral primary sensorimotor cortex and contralateral anterior lobe of the cerebellum, is correlated with a good prognostic outcome for patients in the slow recovery group. A high degree of connectivity between supplementary motor areas, bilaterally, appears to influence functional recovery adversely. PMID- 12546355 TI - Determination of histopathological tumor grade in neuroepithelial brain tumors by using spectral pattern analysis of in vivo spectroscopic data. AB - OBJECT: In this study, 1H magnetic resonance (MR) spectroscopy was prospectively tested as a reliable method for presurgical grading of neuroepithelial brain tumors. METHODS: Using a database of tumor spectra obtained in patients with histologically confirmed diagnoses, 94 consecutive untreated patients were studied using single-voxel 1H spectroscopy (point-resolved spectroscopy; TE 135 msec, TE 135 msec, TR 1500 msec). A total of 90 tumor spectra obtained in patients with diagnostic 1H MR spectroscopy examinations were analyzed using commercially available software (MRUI/VARPRO) and classified using linear discriminant analysis as World Health Organization (WHO) Grade I/II, WHO Grade III, or WHO Grade IV lesions. In all cases, the classification results were matched with histopathological diagnoses that were made according to the WHO classification criteria after serial stereotactic biopsy procedures or open surgery. Histopathological studies revealed 30 Grade I/II tumors, 29 Grade III tumors, and 31 Grade IV tumors. The reliability of the histological diagnoses was validated considering a minimum postsurgical follow-up period of 12 months (range 12-37 months). Classifications based on spectroscopic data yielded 31 tumors in Grade I/II, 32 in Grade III, and 27 in Grade IV. Incorrect classifications included two Grade II tumors, one of which was identified as Grade III and one as Grade IV; two Grade III tumors identified as Grade II; two Grade III lesions identified as Grade IV; and six Grade IV tumors identified as Grade III. Furthermore, one glioblastoma (WHO Grade IV) was classified as WHO Grade I/II. This represents an overall success rate of 86%, and a 95% success rate in differentiating low-grade from high-grade tumors. CONCLUSIONS: The authors conclude that in vivo 1H MR spectroscopy is a reliable technique for grading neuroepithelial brain tumors. PMID- 12546356 TI - The natural history of hemangioblastomas of the central nervous system in patients with von Hippel-Lindau disease. AB - OBJECT: The goals of this study were to define the natural history and growth pattern of hemangioblastomas of the central nervous system (CNS) that are associated with von Hippel-Lindau (VHL) disease and to correlate features of hemangioblastomas that are associated with the development of symptoms and the need for treatment. METHODS: The authors reviewed serial magnetic resonance images and clinical histories of 160 consecutive patients with VHL disease who harbored CNS hemangioblastomas and serially measured the volumes of tumors and associated cysts Six hundred fifty-five hemangioblastomas were identified in the cerebellum (250 tumors), brainstem (64 tumors, all of which were located in the posterior medulla oblongata), spinal cord (331 tumors, 96% of which were located in the posterior half of spinal cord), and the supratentorial brain (10 tumors). The symptoms were related to a mass effect. A serial increase in hemangioblastoma size was observed in cerebellar, brainstem, and spinal cord tumors as patients progressed from being asymptomatic to symptomatic and requiring surgery (p < 0.0001). Twenty-one (72%) of 29 symptom-producing cerebellar tumors had an associated cyst, whereas only 28 (13%) of 221 nonsymptomatic cerebellar tumors had tumor-associated cysts (p < 0.0001). Nine (75%) of 12 symptomatic brainstem tumors had associated cysts, compared with only four (8%) of 52 nonsymptomatic brainstem lesions (p < 0.0001). By the time the symptoms appeared and surgery was required, the cyst was larger than the causative tumor; cerebellar and brainstem cysts measured 34 and 19 times the size of their associated tumors at surgery, respectively. Ninety-five percent of symptom-producing spinal hemangioblastomas were associated with syringomyelia. The clinical circumstance was dynamic. Among the 88 patients who had undergone serial imaging for 6 months or longer (median 32 months), 164 (44%) of 373 hemangioblastomas and 37 (67%) of 55 tumor associated cysts enlarged. No tumors or cysts spontaneously diminished in size. Symptomatic cerebellar and brainstem tumors grew at rates six and nine times greater, respectively, than asymptomatic tumors in the same regions. Cysts enlarged seven (cerebellum) and 15 (brainstem) times faster than the hemangioblastomas causing them. Hemangioblastomas frequently demonstrated a pattern of growth in which they would enlarge for a period of time (growth phase) and then stabilize in a period of arrested growth (quiescent phase). Of 69 patients with documented tumor growth, 18 (26%) harbored tumors with at least two growth phases. Of 160 patients with hemangioblastomas, 34 patients (median follow up 51 months) were found to have 115 new hemangioblastomas and 15 patients new tumor-associated cysts. CONCLUSION: In this study the authors define the natural history of CNS hemangioblastomas associated with VHL disease. Not only were cysts commonly associated with cerebellar, brainstem, and spinal hemangioblastomas, the pace of enlargement was much faster for cysts than for hemangioblastomas. By the time symptoms appeared, the majority of mass effect-producing symptoms derived from the cyst, rather than from the tumor causing the cyst. These tumors often have multiple periods of tumor growth separated by periods of arrested growth, and many untreated tumors may remain the same size for several years. These characteristics must be considered when determining the optimal timing of screening for individual patients and for evaluating the timing and results of treatment. PMID- 12546357 TI - Surgical management of brainstem hemangioblastomas in patients with von Hippel Lindau disease. AB - OBJECT: Hemangioblastomas of the brainstem constitute 5 to 10% of central nervous system (CNS) tumors in patients with von Hippel-Lindau (VHL) disease. At present, optimal management of brainstem hemangioblastomas associated with VHL disease is incompletely defined. In an attempt to clarify some of the uncertainty about the operative treatment of these lesions and its outcome, the authors reviewed all cases of VHL disease in which resection of brainstem hemangioblastomas was performed at the National Institutes of Health during a 10-year period. METHODS: Twelve consecutive patients with VHL disease (six male and six female patients [mean age 31.7 +/- 9 years; range 15-46 years]) who underwent 13 operations to remove 17 brainstem hemangioblastomas were included in this study (mean follow-up period, 88.4 +/- 37.4 months; range 37-144 months). Serial examinations, hospital charts, magnetic resonance images, and operative records were reviewed. To evaluate clinical course, clinical grades were assigned to each patient before and after surgery. Preoperative neurological function was the best predictor of long-term outcome. In addition, patients who underwent CNS surgeries for hemangioblastomas were more likely to improve or to remain neurologically stable. Tumor or cyst size, the presence of a cyst, or the location of the tumor (intramedullary, extramedullary, or mixed; posterior medullary, obex, or lateral) did not affect outcome. No patient was neurologically worse after brainstem surgery. At long-term follow-up review (mean 88.4 months), only one patient had declined neurologically and this was due to the cumulative neurological effects caused by eight additional hemangioblastomas of the spinal cord and their surgical treatment. CONCLUSIONS: Brainstem hemangioblastomas in patients with VHL disease can be removed safely; they generally should be resected when they become symptomatic or when the tumor has reached a size such that further growth will increase the risks associated with surgery, or in the presence of an enlarging cyst. Magnetic resonance imaging is usually sufficient for preoperative evaluation and presurgical embolization is unnecessary. The goal of surgery is complete resection of the lesion before the patient experiences a disabling neurological deficit. PMID- 12546358 TI - Surgical management of spinal cord hemangioblastomas in patients with von Hippel Lindau disease. AB - OBJECT: Von Hippel-Lindau (VHL) disease is an autosomal-dominant disorder frequently associated with hemangioblastomas of the spinal cord. Because of the slow progression, protean nature, and high frequency of multiple spinal hemangioblastomas associated with VHL disease, the surgical management of these lesions is complex. Because prior reports have not identified the factors that predict which patients with spinal cord hemangioblastomas need surgery or what outcomes of this procedure should be expected, the authors have reviewed a series of patients with VHL disease who underwent resection of spinal hemangioblastomas at a single institution to identify features that might guide surgical management of these patients. METHODS: Forty-four consecutive patients with VHL disease (26 men and 18 women) who underwent 55 operations with resection of 86 spinal cord hemangioblastomas (mean age at surgery 34 years; range 20-58 years) at the National Institutes of Health were included in this study (mean clinical follow up 44 months). Patient examination, review of hospital charts, operative findings, and magnetic resonance imaging studies were used to analyze surgical management and its outcome. To evaluate the clinical course, clinical grades were assigned to patients before and after surgery. Preoperative neurological status, tumor size, and tumor location were predictive of postoperative outcome. Patients with no or minimal preoperative neurological dysfunction, with lesions smaller than 500 mm3, and with dorsal lesions were more likely to have no or minimal neurological impairment. Syrinx resolution was the result of tumor removal and was not influenced by whether the syrinx cavity was entered. CONCLUSIONS: Spinal cord hemangioblastomas can be safely removed in the majority of patients with VHL disease. Generally in these patients, hemangioblastomas of the spinal cord should be removed when they produce symptoms or signs. PMID- 12546359 TI - Comparison of anterior and posterior surgical approaches in the treatment of ventral spinal hemangioblastomas in patients with von Hippel-Lindau disease. AB - OBJECT: Von Hippel-Lindau (VHL) disease is an autosomal-dominant neoplastic syndrome with manifestations in multiple organs, which is evoked by the deletion or mutation of a tumor suppressor gene on chromosome 3p25. Spinal hemangioblastomas (40% of VHL disease-associated lesions of the central nervous system) arise predominantly in the posterior aspect of the spinal cord and are often associated with an intraspinal cyst. Rarely, the tumor develops in the anterior aspect of the spinal cord. Ventral spinal hemangioblastomas are a surgical challenge because of difficult access and because vessels feeding the tumor originate from the anterior spinal artery. The goal of this study was to clarify whether an anterior or posterior surgical approach is better for management of hemangioblastomas of the ventral spinal cord. METHOD: . The authors performed a retrospective analysis of clinical outcomes and findings on magnetic resonance (MR) imaging studies in eight patients (two women and six men with a mean age of 34 +/- 15 years) who underwent resection of ventral spinal hemangioblastomas (nine tumors: five cervical and four thoracic). Two surgical approaches were used to resect these tumors. A posterior approach was selected to treat five patients (laminectomy and posterior myelotomy in four patients and the posterolateral approach in one patient); an anterior approach (corpectomy and arthrodesis) was selected to treat the remaining three patients. Immediately after surgery, the ability to ambulate remained unchanged in patients in whom an anterior approach had been performed, but deteriorated significantly in patients in whom a posterior approach had been used, because of motor weakness (four of five patients) and/or proprioceptive sensory loss (three of five patients). This difference in ambulation, despite significant improvements over time among patients in the posterior access group, remained significant 6 months after surgery. In all cases, MR images revealed complete resection of the tumor and in five patients significant or complete resolution of the intramedullary cyst was demonstrated (present in six of eight patients). CONCLUSIONS: The outcomes of these eight patients with hemangioblastomas of the ventral spinal cord indicate that both immediate and long-term results are better when an anterior approach is selected for resection. PMID- 12546360 TI - The safety and utility of recombinant human bone morphogenetic protein-2 for cranial procedures in a nonhuman primate model. AB - OBJECT: The goal of this study was to evaluate the safety and efficacy of recombinant human bone morphogenetic protein 2 (rhBMP-2) in cranial applications. METHODS: Critical-sized calvarial defects were created bilaterally in four rhesus monkeys, and bilateral rectangular bone flaps were created in six others. Control and rhBMP-2-treated sides were randomly chosen for each animal, and an absorbable collagen sponge was used to deliver the growth factor. Over a 6-month period postoperatively, the animals were serially evaluated for bone healing and adverse BMP-related consequences by using the following methods: computerized tomography (CT) scanning, magnetic resonance (MR) imaging, electroencephalography, histological investigations, and cerebrospinal fluid (CSF) analysis. The critical sized defects for the rhBMP-2-treated and control sides attained 71 +/- 12% and 28 +/- 11% closure, respectively (four animals; p = 0.04). The CT scans demonstrated that the bone flaps treated with rhBMP-2 had complete osteointegration in five of six animals, whereas scans of the untreated bone flaps demonstrated uniformly poor osteointegration with the intact skull. Histological analysis confirmed well-formed bridges of bone on the rhBMP-2 treated sides. No epileptogenic activity was detected in any of the animals, and MR imaging revealed no evidence of adverse effects on the brain parenchyma. Meningitic irritation was not found on postoperative CSF sample analysis. CONCLUSIONS: Treatment of bone flaps and critical-sized cranial defects with rhBMP-2 leads to improved bone formation and osteointegration in nonhuman primates. Initial evaluation of rhBMP-2 appears to indicate a good safety profile for use in cranial procedures in primates. PMID- 12546361 TI - Feasibility of using the potassium titanyl phosphate laser with micromanipulators in robotic neurosurgery: a preliminary study in the rat. AB - OBJECT: Robotic surgery can be used as a novel technology in ultramicrosurgery. A microscopic-manipulator (micromanipulator) system, which has a rigid neuroendoscope and three guiding manipulators, was developed in Japan for less invasive telerobotic neurosurgery. To apply this system in a clinical setting, it is necessary to confirm that it is capable of performing various surgical procedures including cutting, coagulation, and bleeding control. The authors chose the potassium titanyl phosphate (KTP) laser for such procedures. The aim of this paper was to evaluate the feasibility of this system mounted with the KTP laser. METHODS: A prototypical micromanipulator system was tested in rats. Two kinds of in vivo experiments were performed using the KTP laser: coagulation and biopsy. The coagulated lesions were precisely aligned and their maximum depths were proportional to the energy applied during the coagulation experiment. The diagnosable specimens were obtained during the biopsy experiment. The micromanipulator system was able to perform all surgical procedures accurately. There was no complication relating to the use of the micromanipulator system such as brain injury or uncontrollable bleeding. CONCLUSIONS: The results from this study proved that this system works precisely and safely and will become a new neurosurgical tool in managing lesions that are difficult to treat using conventional microsurgery or neuroendoscopic surgery. PMID- 12546362 TI - Progressive and extensive dopaminergic degeneration induced by convection enhanced delivery of 6-hydroxydopamine into the rat striatum: a novel rodent model of Parkinson disease. AB - OBJECT: A striatal dopamine lesion induces progressive nigral degeneration in rodents; however, intrastriatal injection of 6-hydroxydopamine (6-OHDA) causes only limited lesions due to spontaneous regeneration of the neurons that survive. To make an extensive lesion, the authors used a convection-enhanced delivery (CED) method for intrastriatal infusion of 6-OHDA and evaluated the animals for a model of Parkinson disease (PD). METHODS: Different doses of 6-OHDA were infused into the unilateral striatum in rats by using the CED method. The dopaminergic neuronal degeneration was evaluated based on morphological, biochemical, and behavioral measurements until 8 weeks postlesion. Due to the wide distribution of the drug, CED of 20 microg of 6-OHDA into the striatum was sufficient to obtain a progressive and extensive nigrostriatal lesion as defined by morphological (> 80% cell loss in the substantia nigra [SN]) and biochemical (> 95% decrease in striatal dopamine) criteria. The extent of the lesion manifested as a stable turning behavior with amphetamine (> 6 turns/minute) and apomorphine (> 4 turns/minute). It also appeared that at I week postlesion the apoptotic markers were maximal in neurons of the SN. CONCLUSIONS: A rat model of PD with a progressive and extensive dopamine lesion was successfully made by intrastriatal CED of 6-OHDA. In this model, the therapeutic value can be assessed using behavioral, biochemical, and histochemical measurements. The delay of nigral neuronal death with respect to the time of 6-OHDA administration may provide a therapeutic window for testing neuroprotective strategies. PMID- 12546363 TI - Characterization of the levels of expression of retinoic acid receptors, galectin 3, macrophage migration inhibiting factor, and p53 in 51 adamantinomatous craniopharyngiomas. AB - OBJECT: Craniopharyngiomas are histopathologically defined as benign tumors that can behave very aggressively at the clinical level. They can originate from different types of embryonal epithelial tissue in which correct spatiotemporal regulation has been disrupted at the effector production level. The goal of this study was to determine the efficacy of using selected biological markers to distinguish between recurring and nonrecurring craniopharyngiomas. METHODS: The authors used computer-assisted microscopy to determine quantitatively the immunohistochemical levels of expression of selected markers, including retinoic acid receptors (RARs), as response elements to retinoic acid in a series of 51 adamantinomatous craniopharyngiomas. These tumors may also originate as the result of physiological defects in the apoptosis-mediated elimination of embryological remnants of epithelial tissue. Galectin-3, p53, and the macrophage migration inhibiting factor (MIF) are known to play crucial roles in these processes. The authors quantitatively determined the levels of expression of these substances in this series of 51 craniopharyngiomas. The data show that all craniopharyngiomas were immunoreactive for RARalpha, whereas their immunoreactivity for RARbeta and RARgamma varied dramatically from one case to another. Craniopharyngiomas with low levels of RARbeta and high levels of RARgamma are more likely to recur than those with higher levels of RARbeta and lower levels of RARgamma. Rapidly recurring craniopharyngiomas also show significantly lower levels of expression of galectin-3 and MIF than nonrecurring or slowly recurring cases. Few tumors exhibited p53 immunopositivity. CONCLUSIONS: The data indicate that even in the so-called adamantinomatous group of craniopharyngiomas, several subgroups with different clinical behavior patterns can be identified on the basis of differentiation markers relating mainly to the presence or absence of RARbeta and RARgamma. PMID- 12546364 TI - Growth inhibition of human malignant glioma cells induced by the PI3-K-specific inhibitor. AB - OBJECT: The phosphatase and tensin homolog deleted from chromosome 10 (PTEN) functions as a tumor suppressor by negatively regulating the growth/survival signals of the phosphatidylinositol 3-kinase (PI3-K)/Akt pathway. The PI3-K/Akt pathway in PTEN-deficient tumors may be one of the key targets for anticancer therapy. The authors examined the effects of the PI3-K inhibitor 2-(4 morpholinyl)-8-phenylchromone (LY294002) on human malignant glioma cells, and compared these effects on PTEN-deficient cells with those on PTEN-wild-type (PTEN wt) cells. METHODS: Using human malignant glioma cell lines, including the PTEN deficient cells A172 and U87MG and the PTEN-wt cells LN18 and LN229, the effects of LY294002 on cell growth, apoptosis, and chemotherapeutic agent-induced cytotoxicity were evaluated. The LY294002 inhibited the growth of U87MG cells associated with reduced phosphatidylinositol 3,4,5,-trisphosphate and phosphorylated Akt, and also induced growth inhibition in three other cell lines. Although LY294002 caused apoptosis in all four cell lines, apoptosis seemed to contribute to only a small portion of growth inhibition induced by LY294002. There was no link between the status of PTEN and the median inhibitory concentration values for LY294002 or between the gene status and the extent of LY294002-induced apoptosis. The LY294002 significantly augmented the cytotoxicity induced by etoposide in PTEN-deficient cells, but not in PTEN-wt cells. Enhancement of 1,3-bis(2-chloroethyl)-1-nitrosourea- and cisplatin-induced cytotoxicity by LY294002 was not linked to the status of PTEN. No marked difference in the amounts of phosphorylated Akt was found between PTEN-deficient and PTEN-wt cells. CONCLUSIONS: The findings show that PI3-K is a possible target for therapy in patients with gliomas, and PI3-K inhibitors in combination with chemotherapeutic agents could be potent therapeutic modalities for patients with malignant gliomas. PMID- 12546365 TI - Comparative genomic hybridization analysis of craniopharyngiomas. AB - OBJECT: Craniopharyngioma is the most common childhood brain tumor and is thought to arise from embryonic remnants of the Rathke pouch. Some craniopharyngiomas are monoclonal in origin and hence presumably harbor somatic genetic alterations, although the precise molecular mechanisms involved in craniopharyngioma development are unknown. The goal of this study was to identify genetic alterations in craniopharyngiomas. METHODS: To gain insight into the molecular mechanisms involved in development of these tumors, the authors analyzed nine adamantinomatous craniopharyngiomas by using comparative genomic hybridization. Six tumors (67%) displayed at least one genomic alteration, and three had six or more alterations. Only two tumors displayed a decrease in DNA copy number, and in all others an increase in DNA copy number was noted. CONCLUSIONS: The authors conclude that a subset of craniopharyngiomas consists of monoclonal tumors arising from activation of oncogenes located at specific chromosomal loci. PMID- 12546366 TI - Intracranial Rosai-Dorfman disease treated with microsurgical resection and stereotactic radiosurgery. Case report. AB - Sinus histiocytosis or Rosai-Dorfman disease (RDD) is a rare idiopathic histioproliferative disorder typically characterized by painless cervical lymphadenopathy, fever, and weight loss. Extranodal, intracranial disease is uncommon. In this report the authors describe the first case of intracranial RDD treated with stereotactic radiosurgery after resection. This 52-year-old man with known RDD presented with a 7-day course of fever, headache, diplopia, left facial paresthesias, and difficulty swallowing. No cranial nerve deficits were evident on examination, but right submandibular and inguinal node enlargements were noted. On neuroimaging, the patient was found to have a homogeneously contrast enhancing petroclival lesion with extension into the left cavernous sinus. The patient underwent a combined left petrosal craniotomy and partial labyrinthectomy with duraplasty for biopsy sampling and partial microsurgical resection of the lesion. Microscopic examination of the biopsy specimen revealed the presence of a mixed cellular population with predominant mature histiocytes consistent with RDD. The residual tumor was treated with stereotactic radiosurgery 2 months after resection. On follow-up imaging the lesion had regressed significantly, with only slight dural enhancement remaining. Microsurgical resection for histological diagnosis, followed by stereotactic radiosurgery for residual tumor represents one treatment alternative in the management of intracranial RDD in which a complete resection carries the potential for excess morbidity. PMID- 12546367 TI - Capillary hemangioma of the cavernous sinus. Report of two cases. AB - Capillary hemangiomas are rare benign vascular tumors that tend to occur in children. Whereas the majority of hemangiomas may regress spontaneously, those associated with functional sequelae or severe symptoms may require treatment. Two patients with capillary hemangiomas of the cavernous sinus that caused neurological symptoms were treated with fractionated stereotactic radiation therapy. Both hemangiomas had shown a progressive increase in size during observation before radiation therapy; both tumors regressed after radiotherapy. Up to the time of the last follow-up evaluation both patients experienced symptomatic relief after radiation. One patient's tumor remains in complete remission and the second tumor continues to demonstrate minor residual contrast enhancement without progression. The authors conclude that fractionated stereotactic radiation therapy is a useful treatment modality in the management of symptomatic capillary hemangiomas when these tumors arise in regions of the brain or skull base in which a complete resection cannot be accomplished. PMID- 12546368 TI - Motor cortex stimulation in a patient with intractable complex regional pain syndrome type II with hemibody involvement. Case report. AB - The authors describe the effectiveness of motor cortex stimulation (MCS) in a patient with complex regional pain syndrome (CRPS) Type II, formerly known as causalgia, with hemibody allodynia. During MCS, a subjective sensation of warm paresthesia developed in the painful hand and forearm and spread toward the trunk. Pain and allodynia in the areas associated with this sensation were alleviated significantly. The analgesic effect of stimulation proved to be long lasting and was still present at the 12-month follow up. The authors speculate that MCS might exert its effect through the modulation of thalamic activity in this particular case of CRPS with hemisensory deficit. A central mechanism associated with functional disturbance in noxious-event processing in the thalamus might have an important role in the pathogenesis of the condition. PMID- 12546369 TI - Barotrauma as a possible cause of aneurysmal subarachnoid hemorrhage. Case report. AB - The authors report the case of a 47-year-old man who suffered a diving accident. After regaining consciousness he experienced severe headache. He was initially treated for barotrauma, but the persistent headache led to diagnostic imaging that revealed an aneurysmal subarachnoid hemorrhage. To the authors' knowledge, this is the first report of a ruptured brain aneurysm associated with barotrauma. PMID- 12546370 TI - Apoplectic lymphocytic hypophysitis. Case report. AB - Pituitary apoplexy has not been reported in the setting of lymphocytic hypophysitis. The authors present the case of a woman with sudden vision loss and headache. Magnetic resonance imaging revealed focal hemorrhages in a sellar lesion with suprasellar extension. The results of histopathological analysis were consistent with lymphocytic hypophysitis. PMID- 12546371 TI - Intracranial hypertrophic calcification complicating neuroendoscopy. Report of three cases. AB - Three cases are presented in which progressive hypertrophic calcification formed following ventricular endoscopy. After a ventricular endoscopy has been performed, it has been the authors' practice to seal the burr hole with bone dust. They believe that the calcifications formed from bone dust that fell into the track left by the endoscope. This is the first time this complication has been described. PMID- 12546372 TI - Development of multiple cerebral arteriovenous malformations documented in an adult by serial angiography. Case report. AB - A 50-year-old woman with a parietal intracerebral hematoma was initially treated by hematoma evacuation. Initial preoperative and follow-up angiograms obtained 6 months later demonstrated no pial arteriovenous malformations (AVMs). She suffered a subarachnoid hemorrhage 8 years later. Results of follow-up cerebral angiography revealed the development of previously undetected multiple cerebral AVMs. This appears to be the first reported case of the development of multiple cerebral AVMs in an adult, demonstrated on serial angiography. PMID- 12546373 TI - Histologically benign metastatic meningioma: morphological and cytogenetic study. Case report. AB - The authors report on a 75-year-old man with histologically benign fibroblastic meningioma metastasizing to the lung, liver, spleen, and kidney. The original tumor exhibited a complex karyotype involving different structural and numerical anomalies associated with monosomy of chromosome 22. The implication of chromosome 1p36 was confirmed by fluorescence in situ hybridization in most interphase nuclei. Metastases occurred 4 months after incomplete resection with prior therapeutic embolization. The recurrent tumor in turn displayed anaplastic features and an increased Ki-67 labeling index. Genetic alterations in such morphologically benign meningiomas have been implicated in the malignant development and progression of these tumors. PMID- 12546374 TI - Histopathological findings in a human carotid artery after stent implantation. Case report. AB - Despite the high technical success rates and the acceptable morbidity and mortality rates that are associated with carotid artery (CA) stent implantation, morphological changes in the vessel wall after this procedure are still unknown. The authors describe histopathological findings of the CA after stent implantation in a human autopsy specimen. A 75-year-old man with asymptomatic CA stenosis underwent successful CA stent implantation, but died 8 months afterward of complications surrounding treatment of a thoracic aortic artery aneurysm. The stented CA was isolated from the autopsy specimen and was analyzed macroscopically and with the aid of scanning electron microscopy and histopathological methods. Although there was focal persistence of a chronic inflammatory reaction around the stent struts, a stabilized neointima overlying the stent, which was principally composed of cells that stained positively for alpha-smooth muscle actin and collagen deposition with complete reendothelialization of the luminal surface, was confirmed in the internal CA. In contrast, residual mural thrombus around the stent struts in the common CA led to a concern about the potential risk for distal embolism in the future. The present case provides supportive evidence of stabilized neointima overlying the stent and valuable information regarding morphological characteristics useful for selecting a stent of an appropriate size. PMID- 12546375 TI - Quantitative pupillometry, a new technology: normative data and preliminary observations in patients with acute head injury. Technical note. AB - The authors prospectively used a new hand-held point-and-shoot pupillometer to assess pupillary function quantitatively. Repetitive measurements were initially made in more than 300 healthy volunteers ranging in age from 1 to 87 years, providing a total of 2,432 paired (alternative right eye, left eye) measurements under varying light conditions. The authors studied 17 patients undergoing a variety of nonintracranial, nonophthalmological, endoscopic, or surgical procedures and 20 seniors in a cardiology clinic to learn more about the effects of a variety of drugs. Additionally, the authors carried out detailed studies in 26 adults with acute severe head injury in whom intracranial pressure (ICP) was continuously monitored. Finally, five patients suffering from subarachnoid hemorrhage were also studied. Quantitative pupillary measurements could be reliably replicated in the study participants. In healthy volunteers the resting pupillary aperture averaged 4.1 mm and the minimal aperture after stimulation was 2.7 mm, resulting in a 34% change in pupil size. Constriction velocity averaged 1.48 +/- 0.33 mm/second. Pupillary symmetry was striking in both healthy volunteers and patients without intracranial or uncorrected visual acuity disorders. In the 2,432 paired measurements in healthy volunteers, constriction velocity was noted to fall below 0.85 mm/second on only 33 occasions and below 0.6 mm/second on eight occasions (< one in 310 observations). In outpatients, the reduction in constriction velocity was observed when either oral or intravenous narcotic agents and diazepam analogs were administered. These effects were transient and always symmetrical. Among the 26 patients with head injuries, eight were found to have elevations of ICP above 20 mm Hg and pupillary dynamics in each of these patients remained normal. In 13 patients with a midline shift greater than 3 mm, elevations of ICP above 20 mm Hg, when present for 15 minutes, were frequently associated with a reduction in constriction velocity on the side of the mass effect to below 0.6 mm/second (51% of 156 paired observations). In five patients with diffuse brain swelling but no midline shift, a reduction in constriction velocities did not generally occur until the ICP exceeded 30 mm Hg. Changes in the percentage of reduction from the resting state following stimulation were always greater than 10%, even in patients receiving large doses of morphine and propofol in whom the ICP was lower than 20 mm Hg. Asymmetry of pupillary size greater than 0.5 mm was observed infrequently (< 1%) in healthy volunteers and was rarely seen in head-injured patients unless the ICP exceeded 20 mm Hg. Pupillometry is a reliable technology capable of providing repetitive data on quantitative pupillary function in states of health and disease. PMID- 12546376 TI - Noninvasive intracranial compliance monitoring. Technical note and clinical results. AB - Although invasive measurement of intracranial pressure (ICP) involving high resolution waveform analysis allows assessment of intracranial compliance (ICC), it is only feasible in a few selected neurosurgical conditions. Intracranial compliance can be assessed using the high-frequency centroid (HFC), which is the power-weighted mean frequency within the 4 to 15-Hz band of the ICP waveform. The authors have systematically tested the utility, performance, and reliability of a noninvasive monitor of ICC. The underlying principle of this device is that the ICP transmission and its infrasonic waves are transmitted through the inner ear toward the tympanic membrane. If the outer ear is sealed in an airtight fashion, motions of the tympanic membrane cause air pressure fluctuations that can be recorded using a special sensor. The authors compared the HFC calculated from an intraparenchymal ICP sensor with that obtained simultaneously from an ipsilaterally placed noninvasive device during half of a respiratory cycle (peak to baseline) as well as for three random samples of three heart cycles. They analyzed 32 sessions in 13 patients in whom mechanical ventilation had been established. In four (11%) of 36 sessions they could not demonstrate an adequate signal. For the peak-to-baseline cycle, the mean invasively recorded HFC was 8.05 +/- 0.55 Hz (range 6.7-9 Hz) whereas the mean noninvasively recorded HFC was 8.04 +/- 0.49 Hz (range 7-9.3 Hz). The ICP was 8.5 +/- 5 mm Hg (range 2-24 mm Hg). For the three heart cycles randomly sampled, the values were 7.73 +/- 0.51 Hz (range 6.7-8.6 Hz) and 7.76 +/- 0.56 mm Hg (range 6.5-8.8 mm Hg), respectively. This device allows noninvasive assessment of ICC based on the HFC waveform analysis that is equivalent to that obtained by invasive intraparenchymal recording. The monitoring device may become a valuable tool for monitoring parameters in patients in whom placement of an intracranial sensor is not feasible but assessment of ICC as an alternative to ICP measurement is desired. PMID- 12546377 TI - Intracranial extradural schwannoma of the 12th cranial nerve. Case illustration. PMID- 12546378 TI - Hyperoxia: good or bad? PMID- 12546379 TI - Resection and prognosis. PMID- 12546380 TI - Secondary cerebellar hemorrhage. PMID- 12546381 TI - Ventral correction of postsurgical cervical kyphosis. AB - OBJECT: Cervical kyphotic deformation may develop after surgery involving either the ventral or dorsal approach. Regardless of the cause, the development of a cervical kyphotic deformity should be avoided, if possible, and corrected if present, when appropriate. The authors describe their experience with a technique for the ventral correction of iatrogenic (postoperative) cervical kyphosis. METHODS: A retrospective review of cases involving correction of postoperative iatrogenic cervical kyphosis via an ventral approach was performed. The authors conducted an ventral approach to kyphosis correction. The procedure required specific head positioning (in extension), convergent distraction pins, and an ventrally placed implant (axially dynamic when appropriate) with multiple points of fixation including at least one point of intermediate fixation. The pre- and postoperative sagittal angle and clinical status were evaluated. During a nearly 14-month period, 12 patients met the inclusion criteria. Ten patients underwent a minimum of 6 months of follow up. They comprised the study population. Most patients presented with mechanical neck pain as part of their symptom profile. The mean magnitude of deformity correction (pre- to postoperative) was 20 degrees of lordosis. The mean postoperative sagittal angle was 6 degrees of lordosis. The mean change in the sagittal angle during the follow-up period was 2.2 degrees of lordosis. CONCLUSIONS: The ventral approach to correction of cervical deformity led to the achievement of lordosis in all but one patient. This posture was effectively maintained during the follow-up period. All patients exhibited improvement postoperatively; three experienced complete resolution of their preoperative symptoms. When symptoms are related to postsurgical kyphosis, deformity correction should be considered. Such a procedure may be performed effectively via an ventral approach in most circumstances. PMID- 12546382 TI - Anterior cervical arthrodesis using DOC dynamic stabilization implant for improvement in sagittal angulation and controlled settling. AB - OBJECT: Placing instrumentation in the anterior cervical spine is a common procedure. The bi- and unicortical systems currently available, however, have distinct advantages and disadvantages. The author reports a prospective series in which a new dynamized anterior cervical fixation system was evaluated. METHODS: Thirty-seven patients underwent anterior cervical discectomy and fusion in which the DOC ventral cervical stabilization system was used for indications including cervical spondylotic radiculopathy, disc herniation, trauma, and myelopathy. Patients underwent anterior cervical discectomy and interbody fusion and/or corpectomy. Preoperative and postoperative clinical data included assessment of spinal cord and nerve root deficit, function, neck pain, and arm pain. Preoperative and postoperative radiographic data included sagittal angle, translation, and settling of the graft. Fusion status was determined by the presence of trabecular bridging bone on plain anteroposterior and lateral cervical radiographs. At a mean follow-up time of 1.3 years, postoperative neck or arm pain was resolved in 52% of the patients, restriction on function was mild or absent in 88%, and fusion was successful in 80% of patients and 88% of the treated levels. There was one implant-related complication, one significant dysphagia complication, and a 10.8% donor graft site complication rate. CONCLUSIONS: The use of this system led to a high percentage of pain relief and radiographic fusion. The sagittal angle was controlled while allowing for graft settling. There were no implant failures. PMID- 12546383 TI - Kryorhizotomy: an alternative technique for lumbar medial branch rhizotomy in lumbar facet syndrome. AB - OBJECT: The authors conducted a prospective study to investigate the efficacy of kryorhizotomy, an alternative procedure for lumbar medial branch neurotomy, in the treatment of lumbar facet syndrome (LFS). METHOD: Fifty patients with chronic low-back pain, in whom pain was relieved by controlled diagnostic medial branch blocks of the lumbar zygapophyseal (facet) joints, underwent lumbar medial branch kryorhizotomy. Outcome was evaluated using the Visual Analog Pain Scales and assessment of work capacity. All outcome measures were repeated at 6 weeks, 6 months, and 1 year after surgery. At 1-year follow-up examination, 31 (62%) of 50 patients experienced a good response to lumbar facet kryorhizotomy. Good results with pain relief of 50% or more were obtained in 85% of patients without previous spinal surgery but only in 46% who had undergone previous spinal surgery. This difference was statistically significant. In five patients (16%) in whom a good initial benefit was observed but who experienced increased pain within 6 weeks after kryorhizotomy, the beneficial result was regained after an early repeated procedure. There were no side effects. Overall, 19 (38%) of 50 procedures were not considered successful. In six of these 19 cases a rigid stabilization of the involved segment provided permanent pain relief. CONCLUSIONS: Based on this study, patients with LFS who have not undergone previous spinal surgery benefit significantly from percutaneous lumbar kryorhizotomy. Kryorhizotomy, which has virtually no risk, seems to be a valuable alternative technique to lumbar medial branch neurotomy. PMID- 12546385 TI - Safety of supplemental endplate screws in thoracic pedicle hook fixation. AB - OBJECT: The AO Universal Spine System thoracic pedicle hook design includes a fixation screw that passes obliquely through the inferior facet into the pedicle to engage in the posterior portion of the superior vertebral body endplate. This endplate screw provides additional purchase at the hook-bone interface. To determine the safety of this fixation system the authors reviewed the operative notes, radiographs, and outcomes of patients who underwent placement of endplate screws. METHODS: Thirty-six patients (16 male and 20 female patients) who required posterior thoracic instrumentation for spinal deformity (11 cases), neoplasm (15 cases), and traumatic injury (10 cases) were included in this study. One hundred sixty-four endplate screws were placed (mean 4.3/patient) to augment pedicle hooks for posterior thoracic instrumentation. The number of instrumented levels ranged from seven to 16. The positions of the screws in relation to the pedicle, neural foramen, spinal canal, and endplate were evaluated by assessing plain radiographs and computerized tomography scans (10 cases). Eighty-two screws (56%) were in ideal position. Lateral pedicle wall perforation occurred with 51 screws (35%). Three screws violated the medial wall and nine screws violated the superior or inferior walls of the pedicle. There were no clinical sequelae associated with any of the malpositioned screws. Adequate follow-up radiographic data were not available in five patients. The mean follow-up duration was 19.8 months (range 3-61 months). Two patients required revision surgery at 3 months and 18 months, respectively, because of hook/endplate screw displacement. There was also one case of an endplate screw fracture without hook displacement that was discovered during subsequent revision surgery. The remainder of the endplate screws and associated pedicles hooks maintained their original positions. There was no case of spinal cord, nerve root, pulmonary, or vascular injury. CONCLUSIONS: The placement of supplemental endplate screws in conjunction with thoracic pedicle hooks can be conducted safely. PMID- 12546384 TI - Percutaneous vertebroplasty and kyphoplasty for painful vertebral body fractures in cancer patients. AB - OBJECT: The current North American experience with minimally invasive vertebro- and kyphoplasty is largely limited to the treatment of benign osteoporotic compression fractures. The objective of this study was to assess the safety and efficacy of these procedures for painful vertebral body (VB) fractures in cancer patients. METHODS: The authors reviewed a consecutive group of cancer patients (21 with myeloma and 35 with other primary malignancies) undergoing vertebro- and kyphoplasty at their institution. Ninety-seven (65 vertebro- and 32 kyphoplasty) procedures were performed in 56 patients during 58 treatment sessions. The mean patient age was 62 years (+/- 13 years [standard deviation]) and the median duration of symptoms was 3.2 months. All patients suffered intractable spinal pain secondary to VB fractures. Patients noted marked or complete pain relief after 49 procedures (84%), and no change after five procedures (9%); early postoperative Visual Analog Scale (VAS) pain scores were unavailable in four patients (7%). No patient was worse after treatment. Reductions in VAS pain scores remained significant up to 1 year (p = 0.02, Wilcoxon signed-rank test). Analgesic consumption was reduced at 1 month (p = 0.03, Wilcoxon signed-rank test). Median follow-up length was 4.5 months (range 1 day-19.7 months). Asymptomatic cement leakage occurred during vertebroplasty at six (9.2%) of 65 levels; no cement extravasation was seen during kyphoplasty. There were no deaths or complications related to the procedures. The mean percentage of restored VB height by kyphoplasty was 42 +/- 21%. CONCLUSIONS: Percutaneous vertebro- and kyphoplasty provided significant pain relief in a high percentage of patients, and this appeared durable over time. The absence of cement leakage-related complications may reflect the use of 1) high-viscosity cement; 2) kyphoplasty in selected cases; and 3) relatively small volume injection. Precise indications for these techniques are evolving; however, they are safe and feasible in well selected patients with refractory spinal pain due to myeloma bone disease or metastases. PMID- 12546386 TI - Balloon kyphoplasty: one-year outcomes in vertebral body height restoration, chronic pain, and activity levels. AB - OBJECT: The authors assessed the safety of balloon kyphoplasty in the reduction and repair of osteopenic vertebral compression fractures and report functional outcomes (back pain and activity levels) in the first 96 patients (with 133 fractures) at their institution. Additionally they provide radiographic outcomes in the first 26 patients (41 fractures) treated and followed for 1 year. METHODS: The authors conducted a retrospective chart review of functional outcomes and evaluated radiographs obtained at 1 week, 1 month, 3 months, 6 months, and 1 year postoperatively. CONCLUSIONS: Balloon kyphoplasty safely increases vertebral body height, decreases chronic back pain, and quickly returns geriatric patients to higher activity levels, leading to increased independence and quality of life. PMID- 12546387 TI - Inclination of the odontoid process in the pediatric Chiari I malformation. AB - OBJECT: The quantitative analysis of odontoid process angulation has had scant attention in the Chiari I malformation population. In this study the authors sought to elucidate the correlation between posterior angulation of the odontoid process and patients with Chiari I malformation. METHODS: Magnetic resonance images of the craniocervical junction obtained in 100 children with Chiari I malformation and in 50 children with normal intracranial anatomy (controls) were analyzed. Specific attention was focused on measuring the degree of angulation of the odontoid process and assigning a score to the various degrees. Postoperative outcome following posterior cranial fossa decompression was then correlated to grades of angulation. Other measurements included midsagittal lengths of the foramen magnum and basiocciput, the authors' institutions' previously documented pB-C2 line (a line drawn perpendicular to one drawn between the basion and the posterior aspect of the C-2 body), level of the obex from a midpoint of the McRae line, and the extent of tonsillar herniation. Higher grades of odontoid angulation (retroflexion) were found to be more frequently associated with syringomyelia and particularly holocord syringes. Higher grades of angulation were more common in female patients and were often found to have obices that were caudally displaced greater than three standard deviations below normal. CONCLUSIONS: These results not only confirm prior reports of an increased incidence of a retroflexed odontoid process in Chiari I malformation but quantitatively define grades of inclination. Grades of angulation were not found to correlate with postoperative outcome. It is the authors' hopes that these data add to our current limited understanding of the mechanisms involved in hindbrain herniation. PMID- 12546388 TI - Enhanced primary stability through additional cementable cannulated rescue screw for anterior thoracolumbar plate application. AB - OBJECT: The authors conducted a study to investigate the biomechanical in vitro influence of a new anchorage system for fixation of anterior stabilization devices and the possibility of using additional cement after screw insertion to compensate for poor bone quality. The incidence of osteoporosis-related fractures has increased nearly twofold in the last decade. Because of problems associated with anterior screw fixation such as loosening, mechanical failure, and the weakness of osteoporotic bone, current surgical treatments of vertebral body (VB) fractures are problematic. This is due to poor fixation strength of anterior screws in the adjacent segments. The aim of this study was to determine whether a new cemented and uncemented VB screw provides improved primary stability following placement of anterior instrumentation in cases of fracture. METHODS: The primary stability-related parameters of a new uncemented/cemented screw were compared with those of conventional monocortical screw fixation in a burst fracture model in which strut graft and anterior overbridging instrumentation were used. The use of the new uncemented screw improved the range of motion (ROM) of the stabilized spine in flexion-extension by approximately 22%, in rotation by 20%, and in lateral bending by 15%. Additional cementation improved the ROM by approximately 41% in flexion-extension, 32% in rotation, and 30% in lateral bending compared with conventional monocortical screw fixation. CONCLUSIONS: The new cannulated screw improves fixation strength and primary stability parameters. It is useful in the initial treatment of fractures in cases of poor bone quality and as a rescue device if previously inserted screws do not remain securely in place. PMID- 12546389 TI - Evaluation of time-dependent spread of tissue damage in experimental spinal cord injury by killed-end evoked potential: effect of high-dose methylprednisolone. AB - OBJECT: Histopathological studies on spinal cord injury (SCI) have demonstrated time-dependent spread of tissue damage during the initial several hours postinjury. When the long tract within the spinal cord is stimulated, a large monophasic positivity occurs at the injury site. This type of potential, termed the killed-end evoked potential (KEEP), indicates that a nerve impulse approaches but does not pass beyond the injury site. The authors tested the hypothesis that the damage spread can be evaluated as a progressive shift of the KEEP on a real time basis. The effect of high-dose methylprednisolone sodium succinate (MPSS) on the spread of tissue damage was also examined by this methodology. METHODS: The KEEP was recorded using an electrode array placed on the spinal cord at the T-10 level in cats. This electrode array consisted of multiple 0.2-mm-diameter electrodes, each separated by 0.5 mm. Spinal cord injury was induced using a vascular clip (65 g pinching pressure for 30 seconds). The midline posterior surface of the spinal cord was stimulated bipolarly at the C-7 level by applying a single pulse at supramaximal intensity. During the initial period of 6 hours postinjury, the localization of the largest KEEP shifted progressively up to 2.5 mm rostral from the injury site. The amplitude of the KEEP recorded at the injury site decreased to 55 to 70% and became slightly shortened in latency as the localization of the largest KEEP shifted rostrally. These findings imply that the injury site KEEP represents the volume-conducted potential of the largest KEEP at the site of the conduction block. It moved away from the injury site in association with the damage spread, and this was confirmed histopathologically. A decrease in amplitude of KEEP at the injury site appeared to be the most sensitive measure of the damage spread, because the amplitude of the volume conducted KEEP is inversely proportional to the square of the distance between the recording site and site of conduction block. Administered immediately after SCI, MPSS clearly inhibited these events, especially within 30 minutes postinjury. CONCLUSIONS: The KEEP enables sequential evaluation to be made of the time-dependent spread of tissue damage in SCI in the same animal. It is, therefore, useful for detecting the effect of therapeutic interventions and for determining the therapeutic time window. The efficiency of MPSS to inhibit the spread of damaged tissue appeared to be maximized when it was administered within the initial 30-minute period postinjury. PMID- 12546390 TI - Age-related changes in expression of transforming growth factor-beta and receptors in cells of intervertebral discs. AB - OBJECT: The authors conducted a study to determine age-related changes in expression of transforming growth factor (TGF)-beta1, -beta2, -beta3, and Type I and Type II receptors in various cells in the nucleus pulposus and anulus fibrosus. Immunolocalization of TGFbetas and Type I and II receptors was examined during the aging process of cervical intervertebral discs in senescence accelerated mice (SAM). The TGFbeta family has important roles for cellular function of various tissues. Its role in disc aging, however, is unknown. Detailed information on the temporal and spatial localization of TGFbetas and their receptors in discs is required before discussing introduction of them clinically into the intervertebral disc. METHODS: Three groups of five SAM each were used. The groups of SAM were age 8, 24, and 50 weeks, respectively. Hematoxylin and eosin staining and immunohistochemical study involving specific antibodies for TGFbeta1, -beta2, -beta3, and Types I and II TGF receptors were performed. Intervertebral discs exhibited degenerative change with advancing age. The TGFbetas and their receptors were present in the fibrocartilaginous cells within the anulus fibrosus and notochord-like cells within the nucleus pulposus of young mice. Expression of TGFbetas and Type I and Type II receptors changed markedly in the cells within the anulus fibrosus during the aging process. CONCLUSIONS: The TGFbetas and their receptors were present in cells within the nucleus pulposus and the anulus fibrosus of young mice, and their expression decreased with age. PMID- 12546391 TI - Intramedullary spinal cavernous malformation following spinal irradiation. Case report and review of the literature. AB - There is a growing body of evidence in the literature suggesting that cavernous malformations of the central nervous system may develop after neuraxis irradiation. The authors discuss the case of a 17-year-old man who presented with progressive back pain and myelopathy 13 years after undergoing craniospinal irradiation for a posterior fossa medulloblastoma. Spinal magnetic resonance (MR) imaging, performed at the time of his initial presentation with a medulloblastoma, demonstrated no evidence of a malformation. Imaging studies and evaluation of cerebrospinal fluid revealed no evidence of recurrence or dissemination. Spinal MR imaging demonstrated an extensive lesion in the thoracic spine with an associated syrinx suggestive of a cavernous malformation. A thoracic laminectomy was performed and the malformation was successfully resected. Pathological examination confirmed the diagnosis. The patient did well after surgery and was ambulating without assistance 6 weeks later. To the best of the authors' knowledge, this is the second reported case in the literature and the first in the young adult age group suggesting the de novo development of cavernous malformations in the spinal cord after radiotherapy. An increased awareness of these lesions and close follow-up examination are recommended in this setting. PMID- 12546392 TI - Posterior atlantoaxial dislocation without fracture. Case report. AB - Posterior dislocation of the atlas onto the axis without related fracture of the odontoid process is a very rare traumatic condition of which five cases have been previously reported. The authors present a sixth case in which management was different from the others. The patient was successfully treated by open reduction of the dislocation and C1-2 transarticular screw fixation. The rarity of the lesion, the differences in diagnostic studies, and the successful treatment by safe intraoperative reduction and fixation are factors of interest in this case. PMID- 12546393 TI - Ossification of the posterior atlantoaxial membrane. Case report. AB - The authors report a case of ossification of the posterior atlantoaxial membrane that led to the development of cervical myelopathy. Computerized tomography and magnetic resonance imaging were helpful in establishing the diagnosis, and decompressive laminectomy may be an appropriate intervention. PMID- 12546394 TI - Transient rotational compression of the vertebral artery caused by herniated cervical disc. Case report. AB - Of the many causes of vertebrobasilar insufficiency (VBI), extrinsic compression of the vertebral artery (VA) is relatively uncommon. A syndrome of VBI caused by extrinsic compression of the VA secondary to head rotation has been termed positional vertebrobasilar ischemia. The authors present a case of transient VBI caused by herniation of a cervical disc. Transcranial Doppler ultrasonography was used preoperatively to confirm the diagnosis and intraoperatively to monitor cerebral perfusion and to confirm that adequate decompression of the VA had been achieved. PMID- 12546395 TI - Congenital spondylolysis of the axis with associated myelopathy. Case report. AB - Cervical spondylolysis is a rare clinical entity and occurs predominantly at the C-6 level. The authors describe a patient with congenital spondylolysis of the axis that caused myelopathy. The patient was a 57-year-old woman with long standing gait disturbance. Plain cervical radiography revealed a radiolucent defect across the pedicle of the axis. Magnetic resonance imaging of the cervical spine in the neutral, flexion, and extension neck positions as well as a computerized tomography myelography in the neutral neck position failed to demonstrate any spinal cord compression. When she rotated her neck, however, the spinal cord was caught between the hypertrophic anterior arch of the atlas and posterior part of the slipped pedicle of the axis on the contralateral side. The spinal cord was transformed into a pear shape. Mechanical injury to the spinal cord seemed to explain her neurological presentation. This is, to the authors' knowledge, the 15th case of axial spondylolysis and the sixth case of spinal cord involvement of the cervical spondylolysis. No cases involving myelopathy secondary to such a unique mechanism have been reported previously in the literature. PMID- 12546396 TI - Alkaptonuria and lumbar disc herniation. Report of three cases. AB - Alkaptonuria is a rare metabolic disease caused by deficiency of homogentisic acid oxidase and characterized by bluish-black discoloration of cartilages and skin (ochronosis). The authors report the cases of three patients with lumbar disc herniation who underwent discectomy and in whom the nucleus pulposus was discovered to be black. Alkaptonuria was diagnosed after discectomy. Discal herniation requiring surgery is unusual in alkaptonuria, with only a few reports. The symptoms in the three patients disappeared after surgery and no symptoms were demonstrated on follow-up examination. PMID- 12546397 TI - Surgical removal of epidural and intradural polymethylmethacrylate extravasation complicating percutaneous vertebroplasty for an osteoporotic lumbar compression fracture. Case report. AB - The authors report the case of patient with a lumbar vertebral body osteoporotic compression fracture who underwent percutaneous transpedicular polymethylmethacrylate (PMMA)-assisted vertebroplasty in whom extravasation of the cement into the spinal canal caused immediate neurological deterioration. Lateral lumbar radiography and computerized tomography scanning demonstrated the presence of intraspinal PMMA. The patient suffered severe low-back pain, left sided sciatica, and profound left L2-4 distribution weakness and numbness. She underwent immediate L-2 laminectomy, the extra- and intradural PMMA was removed, and instrumentation-assisted lateral mass fusion was performed. The patient recovered without incident and her neurological deficit improved. Extravasation of cement into the spinal canal, neural foramen, paraspinal veins, or disc space has been reported in 11 to 73% of percutaneous transpedicular PMMA-assisted vertebroplasty procedures. It is disturbing that more than one group of authors has documented symptomatic spinal canal PMMA extravasation and that the patients were left severely handicapped because of a stated fear that surgery to remove the cement would be difficult and make them worse. The results achieved in this case refute that published notion. It is important to document that decompressive surgery and PMMA removal from the spinal canal are easy and can lead to immediate neurological improvement. With the increasing popularity of percutaneous transpedicular PMMA-assisted vertebroplasty, the authors suspect that more of these cases will be seen. PMID- 12546398 TI - Low-pressure headaches and spinal cord herniation. Case report. AB - Almost 40 cases of spontaneous transdural spinal cord herniation have been reported in the literature. These patients often present with gait disturbance and sensory changes, and their condition is diagnosed as Brown-Sequard syndrome. The pathogenesis of this condition has remained poorly understood. In particular, there is no agreement whether the dural defect is acquired or congenital. In the reported case, a 21-year-old man presented with a 3-year history of intermittent low-pressure headaches consistent with intracranial hypotension. Eventually, the headaches resolved but he developed myelopathy due to a spinal cord herniation. In this case, the authors hypothesize that the progressive spinal cord herniation through a spontaneous dural tear sealed the site of the cerebrospinal fluid leak, causing the resolution of headaches. PMID- 12546399 TI - Intraoperative cervical epidurography: a simple modality for assessing the adequacy of decompression during anterior cervical procedures. Technical note. AB - OBJECT: Intraoperative cervical epidurography (ICE) was used during anterior cervical procedures to assess the success of bone resection and indicate the need for immediate correction in the event of inadequate decompression. METHODS: The adequacy of decompression was assessed by the operating neurosurgeon who performed ICE after anterior microdiscectomy with exposure of the dura mater. If the decompression was deemed inadequate, additional bone was removed using a high speed drill. Epidurography was conducted after each subsequent decompression until adequate bone removal was achieved. This was undertaken in 39 patients undergoing anterior cervical procedures. Nine patients underwent corpectomy, 21 one-level, and nine two-level discectomy. The ICE revealed insufficient resections requiring additional bone removal in 14 (36%) of the 39 patients. CONCLUSIONS: The goal of ICE is to ensure adequate decompression, and if such has not been achieved, to allow for additional immediate bone excision. Furthermore, ICE provides additional real-time control of the position of cages and screws to avoid dural tear or spinal cord compression. Because of this immediate feedback, the success rate of anterior cervical procedures can be improved. The advantages of this modality include real-time assessment, low cost, simplicity, and speed. PMID- 12546400 TI - Guide device for percutaneous placement of translaminar facet screws after anterior lumbar interbody fusion. Technical note. AB - Because the degree of immediate stabilization provided by cage-assisted anterior lumbar interbody fusion (ALIF) has been shown by several studies to be inadequate, supplementary posterior fixation, such as that created by translaminar or transpedicle screw fixation, is necessary. In this study, the authors studied the ALIF-augmentation procedure in which a special guide device is used to place percutaneously translaminar facet screws in 18 patients with degenerative lumbar disease. The minimum follow-up period was 1 month (mean 6 months, range 1-13 months). Degenerative spondylolisthesis with foraminal stenosis was diagnosed in nine patients, associated degenerative disc disease alone or combined with foraminal stenosis in eight, and recurrent disc herniation in one. Following screw placement, computerized tomography scanning was conducted to evaluate the accuracy of the facet screw positioning. All screws were properly placed. No screw penetrated the spinal canal or injured the neural structures. Excellent or good clinical outcomes were demonstrated in all patients at the last follow up. The use of this guide device for post-ALIF percutaneous translaminar facet screw fixation represents a safe, accurate, and minimally invasive modality with which to achieve immediate solid fixation in the lumbar spine. PMID- 12546401 TI - Evaluation of pedicle screw position on computerized tomography scans. Technical note. AB - The quality of a computerized tomography (CT) scan is significantly reduced by metal artifact caused by a pedicle screw system. The purpose of this study was to develop a method of facilitating the evaluation of pedicle screw position on CT scans obtained after screw insertion. The authors developed an algorithm to process spiral CT scans in a personal computer. This uses a digital image enhancement technique, the curve change-based intensity transformation algorithm. This method can generate a clear image of the screw outlines while reducing metal artifact. The resulting images are displayed in arbitrary planes as well as in axial, coronal, and sagittal planes, to support better the evaluation of pedicle screw position. The algorithm was tested using CT scans obtained in 37 patients in whom 186 pedicle screws had been placed. There were five types of screw systems, all of which were made of titanium alloys. In all cases algorithm-based determination of screw position became more convenient and more accurate than when using the conventional bone window setting. In addition, it provided better soft-tissue visualization than the bone window. The software, by displaying clear outlines of screws and decreasing metal artifact, as well as by reconstructing the images in arbitrary planes, was more helpful in identifying the position of pedicle screws than the conventional bone window setting. PMID- 12546402 TI - Malignant optic nerve glioma: metastases to the spinal neuraxis. Case illustration. PMID- 12546403 TI - Solitary cervical fibrous tumor. Case illustration. PMID- 12546404 TI - Neurenteric cyst of the cervical spine with mediastinal extension. Case illustration. PMID- 12546405 TI - Cystic ectopic schwannoma extending anteriorly from the pontomedullary cistern to the thoracic spinal cord. Case illustration. PMID- 12546406 TI - Anterior cervical fusion. PMID- 12546407 TI - Extreme lateral-transatlas approach. PMID- 12546408 TI - Spinal cord vascular lesions. PMID- 12546409 TI - Production of 13C labelled pea flour for use in human digestion and fermentation studies. AB - Stable isotope breath tests offer a new approach to the study of digestion and fermentation of carbohydrates in man. In this study, 13C labelled peas were grown by pulsing 250 ml 13CO2 into a sealed growth chamber. A second pulse was added to a portion of the peas to increase the 13C enrichment. This generated pea flour with an enrichment of 2.36 at.% excess (range 2.09-2.71 n = 3) and 8.64 atom % excess (range 7.37-9.78 n = 3) respectively. This represented incorporation of an absolute yield of 3.8% of the 13CO2 into peas in the 'once-labelled' treatment and 7.5% in the 'twice-labelled' treatment. Ingestion of a mixture of the labelled pea flour (300 mg) by two volunteers generated measurable 13CO2 excretion for breath test analysis. The profile of breath 13CO2 enrichment increased to a maximum within three hours after consuming the pea flour followed by a decrease almost back to baseline by 13 hours. Breath 13CO2 appeared to rise again after this apparent nadir at 13 hours until the end of the sampling period. Mathematical analysis of the data suggested that two peaks best described the profile of breath 13CO2 up to 13 hours. A third peak was necessary to describe the late rise in breath 13CO2 enrichment. This use of 13C enriched pea flour may provide a useful non invasive method for measurement of digestion and fermentation in vivo. PMID- 12546410 TI - Natural carbon isotopes used to study methane consumption and production in soil. AB - Changes in the isotopic composition of carbon can be used to reveal simultaneous occurrence of methane production and oxidation in soil. The method is conducted in laboratory jar experiments as well as in the field by using flux chambers. Simultaneous occurrence of production and oxidation of methane was suggested. PMID- 12546411 TI - Nitrogen-15 fractionation in the thermal decomposition of nitrous oxide of natural isotopic composition. AB - The 15N fractionation in the thermal decomposition of nitrous oxide (N2O) of natural isotopic composition has been investigated in quartz reaction vessel in the temperature interval 888-1073 K. The formulas relating the observed experimentally 15N fractionations with the primary 15N kinetic isotope effect, (k14/k15)p for 14N15N16O, and secondary 15N kinetic isotope effect, (k14/k15)s for 15N14N16O, have been derived. The experimentally estimated 15N kinetic isotope effects have been compared with the primary and secondary 15N kinetic isotope effects calculated with the absolute rate theory formulations applied to linear three atom molecules. A good agreement was found for the primary 15N kinetic isotope effect, (k14/k15)p, in the temperature interval 888-1007 K. But at 1073 K the decompositions of N2O, accompanied by NO (nitric oxide) formation proceed with a twice times smaller primary kinetic isotope effect, (k14/k15)p of 1.0251 +/- 0.0009, only, suggesting the nonlinear transition state structures with participation of the fourth external atom at high temperature decompositions of nitrous oxide. The nitrogen isotope effects determined in this study correlate well with nitrogen isotope fractionations observed in the natural biological, earth and atmospheric processes. PMID- 12546412 TI - Carbon isotope fractionation in the thermal decomposition of uranyl formate. PMID- 12546413 TI - A modified technique for the preparation of SO2 from sulphates and sulphides for sulphur isotope analyses. AB - A modified technique for the conversion of sulphates and sulphides to SO2 with the mixture of V2O5-SiO2 for sulphur isotopic analyses is described. This technique is more suitable for routine analysis of large number of samples. Modification of the reaction vessel and using manifold inlet system allows to analyse up to 24 samples every day. The modified technique assures the complete yield of SO2, consistent oxygen isotope composition of the SO2 gas and reproducibility of delta34S measurements being within 0.10 per thousand. It is observed, however, oxygen in SO2 produced from sulphides differs in delta18O with respect to that produced from sulphates. PMID- 12546414 TI - Carbon isotope analysis in urea at high 13C-abundances using the 13/12CO2-breath test device FANci2. AB - The increasing application of 13C-labelled urea in medicine requires simple and reasonable methods for measuring highly enriched C in urea. The combination: ultimate organic analysis--mass spectrometry so far prescribed is complicated and expensive. For medical diagnosis, however, isotope selective nondispersive infrared spectrometers (NDIRS) have been available for many years. One of these tools is FANci2 which is very reasonable and easily to be operated. By means of such devices also urea highly enriched in 13C can be analysed, provided that the samples are first diluted with a defined amount of urea of natural isotopic composition and then transformed into carbon dioxide by means of urease. The relative abundance of 13C in this carbon dioxide, measured by nondispersive infrared spectrometry, is then a measure of the 13C abundance in the initial urea sample. Comparison of results of such measurements with those attained by mass spectrometry proves that this procedure is feasible and yields precis results. PMID- 12546415 TI - Thielavins as glucose-6-phosphatase (G6Pase) inhibitors: producing strain, fermentation, isolation, structural elucidation and biological activities. AB - High-throughput screening of microbial extracts using rat hepatic microsomal glucose-6-phosphatase (G6Pase) led us to find thielavin B as a G6Pase inhibitor with inhibition of glucose output from glucagon-stimulated hepatocytes. Further searching for more potent analogs identified 11 new thielavins F-P in addition to the known thielavins A and B from a fungus Chaetomium carinthiacum ATCC 46463. Thielavin G showed the strongest activity as a G6Pase inhibitor (IC50=0.33 microM), while the IC50 of thielavin B was 5.5 microM. According to the structure activity relationship, including authentic thielavins C, D and 3 partial hydrolysates from thielavins A and B, 3 benzoic acid-units and carboxylic acid functions are essential for G6Pase inhibition. PMID- 12546416 TI - New antibiotics miyakamides produced by a fungus. AB - New antibiotics, miyakamides A1, A2, B1, and B2, were isolated from the cultured broth of Aspergillus flavus Link var. columnaris FKI-0739 together with known compounds, parasiticolide A, hydroxyaspergillic acid, and kojic acid. The structure of miyakamide A1 is N-acetyl-L-phenylalanyl-N-methyl-L-phenylalanyl (alphaZ)-alpha,beta-didehydrotryptamine, and miyakamide A2 is E isomer of A1 at didehydrotryptamine. The structure of miyakamide B1 is N-acetyl-L-tyrosyl-N methyl-L-phenylalanyl-(alphaZ)-alpha,beta-didehydrotryptamine, and B2 is E isomer of B1. Both miyakamides A1 and B1 existed as equilibrium isomers in solvents, and this isomerism was associated with cis-trans rotation of the amide bond between two amino acids. Conformational isomerism between two amino acids of miyakamides A2 and B2 is cis-form. Miyakamides showed growth inhibitory activity against brine shrimp, Artemia salina. PMID- 12546417 TI - Isolation of temperature-sensitive Saccharomyces cerevisiae with a mutation in erg25 for C-4 sterol methyl oxidase. AB - C-4 sterol methyl oxidase encoded by the ERG25 gene is a key enzyme in the ergosterol biosynthetic pathway in fungi. ERG25p contains three histidine clusters common to nonheme iron binding enzymes and endoplasmic reticulum retrieval signal. In order to characterize ERG25p, we generated a series of temperature-sensitive(ts) erg25 mutants by random mutagenesis. One of the resulting mutants, the mERG25 strain, accumulated 4,4-dimethlzymosterol at the nonpermissive temperature. Sequence analysis of the mERG25 mutant indicated three amino acid substitutions in ERG25p, namely N48D, V133A, and F135S. These results indicate that the ERG25 gene product is a new antifungal target. PMID- 12546418 TI - PF1163A, a novel antifungal agent, inhibit ergosterol biosynthesis at C-4 sterol methyl oxidase. AB - PF1163A and B are a pair of antifungal agents isolated from a fermentation broth of Penicillium sp. PF1163A inhibited ergosterol synthesis in Saccharomyces cerevisiae, resulting in an accumulation of 4,4-dimethylzymosterol and a decrease of ergosterol. The ERG25 strain overexpressing the ERG25 gene was resistant to PF1163A. ERG25p is a C-4 sterol methyl oxidase known to be essential for the viability of yeast and fungi because of the known role of ERG25 gene disruption in S. cerevisiae-led lethality. ERG25p is the enzyme responsible for the first step in the removal of the two methyl groups at the C-4 position of sterol. From the results obtained here, we conclude that PF1163A is a novel natural antifungal that inhibits C-4 sterol methyl oxidase. PMID- 12546420 TI - Synthesis of milbemycins alpha9, alpha10, alpha11, alpha12, alpha14, alpha15, alpha20, alpha21, alpha22, alpha23, alpha26, alpha27, delta(2,3),delta(4,26) milbemycins A3, A4 from milbemycins A3, A4, and their acaricidal activities. AB - Chemical derivation methods to prepare 26-acyloxy and 26-hydroxymilbemycins, which had been reported as natural products, milbemycins alpha9, alpha10, alpha11, alpha12, alpha14, alpha15, alpha20, alpha21, alpha22, alpha23, alpha26, alpha27 from milbemycins A3, A4 were reported. Delta(2,3),delta(4,26)-milbemycins A3, A4, which had also been reported as natural products, were further prepared from milbemycins A3, A4. Their acaricidal activities were also assessed against the organophosphorus-sensitive two-spotted spider mite (Tetranychus urticae) on primary leaves of cowpea plants (Vigna sinesis Savi species) by spraying. PMID- 12546419 TI - S-3578, a new broad spectrum parenteral cephalosporin exhibiting potent activity against both methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. Synthesis and structure-activity relationships. AB - A series of 7-aminothiadiazolylcephalosporins having a 1-(substituted)-1H imidazo[4,5-b]pyridinium group at the C-3' position of the cephem nucleus were synthesized and evaluated for in vitro antibacterial activities. Among the cephalosporins prepared in this study, 7beta-[2-(5-amino-1,2,4-thiadiazol-3-yl) 2(Z)-ethoxyiminoacetamido]-3-[1-(3-methylaminopropyl)-1H-imidazo[4,5-b]pyridinium 4-yl]methyl-3-cephem-4-carboxylate sulfate (S-3578) showed extremely potent broad spectrum activity against both gram-positive bacteria including methicillin resistant Staphylococcus aureus (MRSA) and gram-negative bacteria including Pseudomonas aeruginosa, and good water solubility. PMID- 12546421 TI - Phenylpyropene A and B, new inhibitors of acyl-CoA: cholesterol acyltransferase produced by Penicillium griseofulvum F1959. PMID- 12546422 TI - Identification of 6-prenylindole as an antifungal metabolite of Streptomyces sp. TP-A0595 and synthesis and bioactivity of 6-substituted indoles. PMID- 12546423 TI - Screening of interleukin-2 production inhibitor with mouse thymoma EL4 cells. PMID- 12546424 TI - First identification of Streptomyces genes involved in the biosynthesis of 2 deoxystreptamine-containing aminoglycoside antibiotics--genetic and evolutionary analysis of L-glutamine:2-deoxy-scyllo-inosose aminotransferase genes. PMID- 12546425 TI - Neurodegenerative mechanisms in subacute sclerosing panencephalitis. AB - Subacute sclerosing panencephalitis is caused by persistent brain infection of mutated measles virus, showing inflammation, neuronal loss, and demyelination. We neuropathologically examined six autopsy cases of subacute sclerosing panencephalitis, using in situ nick end-labeling and immunohistochemistry. Both the neurons and glial cells in the cerebral cortex showed immunoreactive nuclei in the nick end-labeling in two cases with disease duration within 2 years, whereas they were confined to the glial cells in the demyelinated cerebral white matter in three cases with disease duration ranging from 2 to 10 years. The nuclei and cytoplasm were immunoreactive for 8-hydroxy-2'-deoxyguanosine and 8 hydroxyguanosine, markers of oxidative damage to DNA and ribonucleic acid, respectively, in the cerebral cortex in three cases with disease duration within 9 years. In contrast, 4-hydroxy-2-nonenal-modified proteins, products of lipid peroxidation, were deposited in the demyelinated white matters in four cases with disease duration longer than 9 years. In three cases with long survival, the expression of glial glutamate transporters was reduced in the cerebral cortex. It is speculated in subacute sclerosing panencephalitis that apoptosis and oxidative stress to DNA can contribute to the early neuronal damage, whereas lipid peroxidation and disturbed glutamate transport may be related to the subsequent neurodegeneration. PMID- 12546426 TI - Contrast sensitivity of patients with severe motor and intellectual disabilities and cerebral visual impairment. AB - We attempt to evaluate the residual visual capacities of nine patients (seven males and two females; age range 4 to 35 years, mean 13.8 +/- 9.98) with cerebral visual impairment coupled with severe motor and intellectual disabilities by their contrast sensitivities to sine-wave gratings. Two methods were used for detecting the occurrence of ocular responses to stimuli: (1) detection of optokinetic nystagmus to drifting sinusoidal gratings by naked-eye observation and electronystagmography and (2) detection of ocular pursuit for a drifting Gabor patch by naked-eye observation. We succeeded in measuring the sensitivities of eight cases. For the remaining one case, only the Gabor method could be applied. Most cases showed low contrast sensitivity in both higher (2 and 4 cycles/degree) and lower (0.125 and 0.25 cycles/degree) spatial frequencies and relatively high contrast sensitivity in the middle (0.5 and 1 cycle/degree) range of spatial frequencies. We conclude that the residual visual capacities of patients with severe motor and intellectual disabilities and cerebral visual impairment can be measured fairly accurately by these behavioral methods. PMID- 12546427 TI - Tourette syndrome associated with body temperature dysregulation: possible involvement of an idiopathic hypothalamic disorder. AB - Tourette syndrome is a neuropsychiatric disorder that holds the potential to afflict the emotional, familial, social, or scholastic performances of patients with Tourette syndrome in day-to-day life functioning. The disorder is today characterized mainly and diagnosed by clinical observations, yet false-negative results obtained in the diagnosis of Tourette syndrome are numerous and well documented. There is still no laboratory or imaging technique available for the diagnosis of Tourette syndrome. This article reports on changes of the ambient thermal perception (38%) and a circadian dysregulation of the body-temperature profile present in Tourette syndrome probands, irrespective of their chronologic age, sex, or comorbid symptoms. An involvement of idiopathic hypothalamic dysfunctions associated with Tourette syndrome is proposed. Such a phenomenon, if substantiated, could lead to a better understanding of Tourette syndrome and the development of unbiased physical diagnostic criteria of Tourette syndrome and potentiate possible production of novel therapeutic possibilities. PMID- 12546428 TI - Idiopathic intracranial hypertension in the pediatric population. AB - The aim of this study was to determine the features of pseudotumor cerebri or idiopathic intracranial hypertension in prepubertal and pubertal children. We retrospectively reviewed patient charts of those 16 years and younger, diagnosed with pseudotumor cerebri/idiopathic intracranial hypertension. Our study group consisted of 27 patients; the mean age was 10.9 years, and there was a male-to female ratio of 13 to 14. In the prepubertal group (n = 13), the male-to-female ratio was 8 to 5; in the pubertal group, (n = 14), the ratio was 5 to 9. Overweight or obesity was found in 16 (59%) patients. Outcome was favorable except for one who remained symptomatic. Pseudotumor cerebri/idiopathic intracranial hypertension in children is rare. Its characteristics differ from adults. We found the prepubertal group to be a distinct group since pseudotumor cerebri/idiopathic intracranial hypertension did not occur predominantly in females and was not associated with obesity. PMID- 12546429 TI - Interleukin-1beta, tumor necrosis factor-alpha, and nitrite levels in febrile seizures. AB - Proinflammatory cytokines (such as interleukin-1beta, tumor necrosis factor alpha) and nitric oxide are known to have both direct and indirect modulating effects on neurons and neurotoxic neurotransmitters released during excitation or inflammation. We measured interleukin-1beta, tumor necrosis factor-alpha, and nitrite levels in the peripheral blood and cerebrospinal fluid of children with febrile seizures and compared our results with those of children with febrile illnesses without seizures. Twenty-nine children with febrile seizure and 15 controls were studied. The mean concentrations of interleukin-1beta and nitrite were significantly increased in the cerebrospinal fluid (P < .01) of the children with febrile seizure. There were no significant changes in serum interleukin 1beta, tumor necrosis factor-alpha, nitrite, and cerebrospinal fluid tumor necrosis factor-alpha levels. Our data support the hypothesis that increased production of interleukin-1beta in the central nervous system or increased diffusion of interleukin-1beta through the blood-brain barrier is involved in the pathogenesis of febrile seizures. PMID- 12546430 TI - Transmantle dysplasia in tuberous sclerosis: clinical features and surgical outcome in four children. AB - In the literature, several malformations of cortical development have been described as additional lesions in tuberous sclerosis complex. Among these lesions, a very large focal cortical dysplasia has peculiar magnetic resonance imaging features: a signal abnormality that extends radially inward toward the lateral ventricle from the pachygyric cortical surface plus a homogeneous clinical picture. Affected patients have early-onset drug-resistant epilepsy and severe developmental delay. We describe the clinical, genetic, neurophysiologic, and neuroradiologic characteristics of four patients affected by tuberous sclerosis and this type of cortical dysplasia these patients are of special interest because they have been operated on for their dysplastic lesions. Total control of seizures has been achieved in the three children who underwent a complete lesionectomy. This result cannot be permanent, however, because of the presence of other cortical tubers which could become epileptogenic. All things considered, our choice was to give these children at least temporary relief from severe epilepsy and possibly support for developmental progression. PMID- 12546431 TI - Cardiomyopathy associated with neurologic disorders and mitochondrial phenotype. AB - Cardiomyopathy and neuromuscular abnormalities may simultaneously coexist and present with defects in mitochondrial DNA and bioenergetic function. We sought to evaluate the relationship between clinical and mitochondrial phenotypes in 28 young patients with both cardiomyopathy and neurologic disorders including seizures, dystonia, ophthalmoplegia, Kearns-Sayre syndrome, Leigh disease, and Friedreich's ataxia. All tissues examined displayed marked defects in respiratory complex activities. Five patients had abundant large-scale mitochondrial DNA deletions and one patient displayed a pathogenic point mutation previously reported with mitochondrial cytopathy. In this cohort, patients with hypertrophic cardiomyopathy displayed a higher incidence of complex I defects, fewer DNA deletions and mitochondrial structural abnormalities and were less often associated with developmental delay phenotype compared with patients with dilated cardiomyopathy. Although structural abnormalities are present in a subset of patients, evaluation of respiratory enzyme activity appears to be most informative whether tissues examined were derived from heart or skeletal muscle. Defects in mitochondrial DNA and bioenergetics are frequently present in children with cardiomyopathy presenting with a variety of neurologic abnormalities and are amenable to biochemical and molecular analysis. PMID- 12546432 TI - Differential functional magnetic resonance imaging language activation in twins discordant for a left frontal tumor. AB - Eight-year-old twins, one with a left frontal tumor and aphasic seizures, the other neurologically normal, underwent serial assessment of expressive language with functional magnetic resonance imaging and neuropsychology. The affected twin showed a significant amount of right hemisphere activation coincident with behavioral deterioration in expressive language and late growth in the tumor. This pattern of language dysfunction and the left language dominance of her co twin suggested that the affected twin was also left dominant for language, and the significance of her right activation is discussed. We postulate that the right hemisphere activation represents a stabilizing mechanism in the context of a developmental and progressive lesion in language cortex rather than language transfer per se. PMID- 12546433 TI - Cervicothoracic extradural arachnoid cyst: possible association with obstetric brachial plexus palsy. AB - The association of cervicothoracic extradural arachnoid cysts and obstetric brachial plexus palsy has not previously been reported. We report two patients with this association. The first patient is a 9-month-old boy with left obstetric brachial plexus palsy that developed bilateral leg weakness at 6 months of age owing to compression of the spinal cord by a C6 to T8 left cervicothoracic extradural arachnoid cyst. The second patient is a 3-year-old girl with bilateral brachial plexus palsy and spastic paraparesis who had magnetic resonance imaging at 3 days of age that showed intraspinal cord injury and a cervicothoracic extradural arachnoid cyst compressing the spinal cord. We believe that the association of cervicothoracic epidural arachnoid cysts and obstetric brachial plexus palsy in these patients was causal and recommend that the possibility of a cervicothoracic epidural arachnoid cyst be considered in patients with brachial plexus palsy and evidence of spinal cord injury. PMID- 12546434 TI - Leukoencephalopathy with bilateral anterior temporal lobe cysts: a further case of this new entity. AB - The introduction of magnetic resonance imaging (MRI) and magnetic resonance spectroscopy into clinical practice has permitted advances in the definition and categorization of unknown leukodystrophies in children. We report a new type of leukodystrophy, defined by particular MRI and clinical findings, in a child with uneventful pre- and perinatal histories, spasticity, severe mental impairment with absence of language, and deafness. Brain MRI showed lobar white-matter signal abnormalities with extensive cysts in the anterior temporal lobe. The results of all metabolic screening tests, including the specific investigations for leukodystrophy, have been negative. Six years of clinical and MRI-magnetic resonance spectroscopy patient follow-up indicate a nonprogressive clinical and magnetic resonance picture. Owing to the striking similarities with the previous eight patients reported, this additional case supports the identification of a new leukodystrophy. PMID- 12546435 TI - Long-term findings on brain magnetic resonance imaging in acute encephalopathy with bilateral striatal necrosis associated with measles. AB - The long-term findings on brain magnetic resonance imaging (MRI) in a 7 10/12 year-old boy with a history of acute encephalopathy with bilateral striatal necrosis following measles at the age of 22 months are described. At the early stage of illness, brain MRI studies revealed bilateral, symmetric basal ganglia lesions, predominant on the globi pallidi, appearing as hyperintense signals on T1- and T2-weighted images. Six years later, follow-up brain MRI studies showed that the bilateral, symmetric lesions on the globi pallidi persisted with low signal on T1- and high signal on T2 weighted images. At present, the patient has some persistent neurologic signs. These findings suggest that both clinical and neuroradiologic findings may persist in children with acute encephalopathy with bilateral striatal necrosis following measles. PMID- 12546436 TI - Intractable epilepsy with ring chromosome 20 syndrome treated with vagal nerve stimulation: case report and review of the literature. AB - We report a case of a 6-year old girl with ring chromosome 20 syndrome whose medically intractable seizures were successfully treated with vagal nerve stimulation therapy. Medically intractable seizures are an expected part of this rare syndrome, and the dramatic improvement in seizure control with vagal nerve stimulation is emphasized. Earlier use of vagal nerve stimulation in similar cases should be considered. PMID- 12546437 TI - Delayed onset of status epilepticus after transient asphyxia in an asymptomatic full-term neonate. AB - A full-term neonate presented with status epilepticus at 12 hours of age after a symptom-free interval following transient asphyxia at birth. Conventional neuroimaging failed to detect structural correlates to support recent injury. However, diffusion-weighted magnetic resonance imaging studies revealed recent ischemic brain injury. Placental examination documented multiple subacute and chronic findings indicative of decreased maternal/fetal perfusion. These antepartum placental abnormalities may have been associated with this child's inability to withstand the stress of a prolonged second stage of labor, resulting in intrapartum asphyxia leading to brain injury. This child's clinical presentation highlights the delayed reperfusion phase after ischemia-induced brain injury. PMID- 12546438 TI - Isolated left oculomotor nerve palsy following measles. AB - A 10-month-old boy was admitted with ptosis on the left eyelid, which rapidly occurred following a disease with rash about 20 days before admission to our hospital. By history, none of the vaccinations had been performed. On physical examination, his vital signs were stable, and he had marasmus. Isolated left oculomotor nerve palsy was diagnosed. Cranial magnetic resonance imaging was normal. Serum IgM antibody to measles virus was positive. Oculomotor nerve palsy markedly improved on the 15th day of follow-up, and complete improvement was noted on the second month of follow-up. To our knowledge, this is the first case of oculomotor nerve palsy following measles. PMID- 12546439 TI - Landau-Kleffner syndrome beginning with stuttering: case report. AB - Landau-Kleffner syndrome is marked by an acquired aphasia in children who have had normal language and motor development. A 3.5-year-old girl was referred to our clinic with stuttering. She was diagnosed as having benign myoclonic epilepsy of infancy at 3.5 months of age and treated with valproic acid. Her electroencephalogram (EEG) returned to normal at the end of the first year. The therapy was stopped after a 2-year seizure-free period. She started to stutter prominently 3 months after the discontinuation of antiepilepsy drugs. She had no verbal agnosia. Her EEG revealed multiple spike and wave discharges. She was diagnosed as having Landau-Kleffner syndrome. Her previous epilepsy history had contributed to her having obtained an EEG in the early period. We suggest that if a child with normal language function starts to stutter, Landau-Kleffner syndrome must be considered in the differential diagnosis. PMID- 12546440 TI - Proton magnetic resonance spectroscopy in a case of subacute sclerosing panencephalitis. AB - Subacute sclerosing panencephalitis is an encephalopathy caused by a persistent measles virus infection. We examined a 13-year-old girl with subacute sclerosing panencephalitis and performed a magnetic resonance spectroscopic study to evaluate the in vivo pathophysiologic abnormality. The results suggested the occurrence of inflammatory processes and glial proliferation prior to neuronal loss even in magnetic resonance imaging (MRI)-negative regions as well as in MRI positive regions. The additional resonance peaks were also detected, but further studies should be performed to determine the precise pathophysiologic mechanisms. Novel in vivo modalities such as spectroscopy would be useful as a tool to measure responses to therapy. PMID- 12546441 TI - Clinical feature of unintended thin corneal flap in LASIK: 1-year follow-up. AB - To purpose of this study was to evaluate complications of unintended thin corneal flap in laser in situ keratomileusis (LASIK), such as visual acuity and myopic regression, at the one year follow-up. We performed a study on 54 eyes, i.e. 27 patients, having LASIK. The eyes were split into two groups, group one, 27 eyes with unintended thin corneal flap with a thickness of 100 microm or less, and as a control group, 27 eyes with a thickness of 110 microm or more. The average corneal flap thicknesses of the two groups were 88.89 +/- 8.07 microm and 132.70 +/- 19.58 microm, respectively. With regard to postoperative complications, there were no statistical differences between the groups for: foreign bodies in aspects of the interface, mild peripheral infiltration, superficial punctuate keratitis, myopic regression and decreased vision. The only complication showing any statistical difference between the two groups was the central corneal opacity. An unintended thin corneal flap, with an intact Bowman's layer, induced no significant postoperative complications. Central corneal opacity was apparent in 4 of the eyes in group one, so may be related with a thin corneal flap. PMID- 12546442 TI - Visual function and quality of life in Korean patients with glaucoma. AB - The relationship between visual acuity, the level of visual field impairment, visual functioning, and the quality of life was evaluated in Korean patients with glaucoma. Forty-three consecutive glaucoma patients from the glaucoma service at Severance Eye and Ear Hospital were included in this study. Each subject underwent a vision-specific functional status questionnaire (VF-14), a modified VF-14 (VF'-14) and general quality-of-life questionnaires of the Medical Outcomes Study 36-Items Short Form Health Survey (SF-36). The visual acuity and visual field measurements with a Humphrey automated perimeter were taken within one month of the initial visit. The mean age of the subjects was 57.9 +/- 17.8. Among these patients, there were 23 males and 20 females. The mean deviation (MD) and the corrected pattern standard deviation (CPSD) by the automated perimeter in the eye with the better visual acuity were -5.87 +/- 5.16 dB and 3.92 +/- 2.74 dB respectively. The VF-14 and SF-36 scores were 41.28 +/- 14.56 and 52.39 +/- 6.61 respectively. There were significant correlations between the VF-14, VF'-14, and the MD of the better eye (p < 0.05). In Korean patients with glaucoma, the VF-14 and VF'-14 have significant relationship with the MD of the better eye in the visual field. PMID- 12546443 TI - Comparison of surgical outcomes between small collagen and chromic catgut implants in deep sclerectomy. AB - In deep sclerectomy, collagen implant has been used to maintain space under the scleral flap. However, the effect of other implants has not been studied. In this retrospective study, we compared surgical outcomes between small collagen and chromic catgut used as implants in deep sclerectomy. Deep sclerectomy was performed on 23 patients (25 eyes) who either had an open angle and a high intraocular pressure (IOP) (> 22 mmHg) in spite of receiving the maximal tolerable medical treatment, or who were intolerant to medications. Our study consisted of 14 patients (15 eyes) in the small collagen group and 8 (9 eyes) in the chromic catgut group. The mean follow-up period was 8.6 +/- 3.3 months in the small collagen group and 4.4 +/- 1.2 months in the chromic catgut group. The mean preoperative IOP was not significantly different between the two groups. The complete success rate of the small collagen group was significantly better than that of the chromic catgut group at the final follow-up when data were analyzed using the Kaplan-Meier survival curve (87% versus 30%; P = .01). As for the qualified success rate, the small collagen group showed significantly better results at the final follow-up (93% versus 80%; P = .046). The mean number of postoperative medications was significantly lower in the small collagen group (0.26 versus 1.10; P < .05). These results confirmed that the use of small collagen implant in deep sclerectomy produced a higher success rate and a lowered need for postoperative medication compared to the use of chromic catgut implant. Because equal sized implants were used, the difference in biochemical properties, particularly the severity of inflammation caused by the implant, is presumed to be the cause of the different surgical outcomes. PMID- 12546444 TI - Collateral vessels in branch retinal vein occlusion. AB - Experiments show that collaterals that develop and maturate in branch retinal vein occlusion (BRVO) are helpful in the drainage of blood from a blocked area to an adjacent area. Laser treatment on the nonperfusion area can reduce the retinal blood inflow that can impair the formation of collateral vessels. Furthermore, if collaterals were accidentally destroyed by laser photocoagulation, leakage and neovascularization would increase. Forty-five patients with BRVO were reviewed retrospectively. Collateral vessels were noted from angiographic analysis in 27 of 45 (60%) patients. Good visual prognosis was noted in the patients with collaterals. Cases with neovascularization underwent laser treatment. In one case, laser photocoagulation was applied to the collateral vessels accidentally, after which the leakage significantly increased on fluorescein angiography. In conclusion, collateral vessels in BRVO have a favorable effect on visual prognosis. Careful laser treatment is recommended to avoid destroying collaterals in BRVO. PMID- 12546445 TI - Management of choroidal neovascularization following laser photocoagulation for central serous chorioretinopathy. AB - Little is known about the natural history and management of choroidal neovascularization (CNV) which developed as a complication of laser photocoagulation for central serous chorioretinopathy (CSC). We experienced two patients with CNV which developed after laser treatment for CSC. Submacular membranectomy was performed on both cases after the confirmation of subretinal CNV with optical coherence tomography. One patient received photodynamic therapy for recurrent CNV. The vision of both patients has been improved over 6 months of follow up. These cases suggest that active intervention, including submacular surgery, improves the visual prognosis of this condition. PMID- 12546446 TI - Mutations of the Norrie gene in Korean ROP infants. AB - The present study was conducted to evaluate if there is a Norrie disease gene (ND gene) mutation involved in the retinopathy of prematurity (ROP), and to identify the possibility of a genetic abnormality that may be linked to the presence of ROP. Nineteen premature Korean infants, with a low birth weight (1500 g or less) or low gestational age (32 weeks or less), were included in the study. Eighteen infants had ROP, and the other did not. Genomic DNA was isolated from the peripheral blood leukocytes of these patients, and all three exons and their flanking areas, all known ND gene mutations regions, were evaluated following amplification by a polymerase chain reaction, but no ND gene mutations were detected. Any disagreement between the relationship of ROP to the ND gene mutation will need to be clarified by further investigation. PMID- 12546447 TI - Clinical effect of early surgery in infantile exotropia. AB - To evaluate the effects of early surgical intervention in infantile exotropia on the motor and sensory functions, we reviewed the records of 17 subjects diagnosed with exotropia before the age of 12 months, receiving surgery before the age of 24 months, with a follow-up period greater than one year, between 1996 and 2000. Of the 17 subjects (6 intermittent, 11 constant), 14 (82%) (6 intermittent, 8 constant) had a final horizontal deviation of <10 PD, with 3 (18%) needing a re operation. Fusion and gross binocularity were developed in 7 (4 intermittent, 3 constant), and 11 (6 intermittent, 5 constant) subjects, respectively. Seven subjects developed stereopsis of 200 seconds or better, and 5 of the 6 with intermittent exotropia (83%) being involved. In conclusion, over 80% of the successful alignments were obtained with surgery before the age of 24 months in infantile exotropia, which was similar to previous studies. Furthermore, early surgical intervention, particularly in the intermittent phase, resulted in more effective sensory function. PMID- 12546448 TI - Measurement of acute pain after eye surgery in children. AB - This study was performed to assess the degree of acute pain in children following eye surgery using a Face Pain Rating Scale (FPRS), with a validity and reliability test employing a Numeric/Word Graphic Rating Scale (NWGRS). The degree of pain was obtained at 2, 4, 6, 8 and 24 hours after surgery using the FPRS and NWGRS. The changes in pain intensities were analyzed using a RM-ANOVA, while the relationship between the FPRS and NWGRS was analyzed by a Pearson coefficient in SPSS/WIN. Two third of the children experienced pain equal to, or greater, than moderate to severe, and about one fifth of the subjects expressed the most severe pain at 2 hours after surgery. At 4 hours after surgery, 95.3% of the children still complained of pain, and 8 hours after surgery, 82.8% of the patients experienced 'a little bit' or 'a little more' pain or discomforts. At 1 day after surgery, only 34.4% of children were free of pain. A high correlation between the FPRS and NWGRS was identified over 5 time-points (.887 < r < .735). The gender, type of the surgery and past operation experience had no effects on the degree of pain. Our results suggest that FPRS is a reliable and valid measurement for acute pain assessment in Korean children following eye surgery, and warrants application in the integrated clinical protocol, including non pharmacological and pharmacological management of acute pediatric pain reduction following surgery. PMID- 12546450 TI - A case of retained wooden foreign body in orbit. AB - A 41-year-old man visited our clinic complaining of esodeviation of the right eye. He had been operated on for corneal laceration 3 years before. One month later, exodeviation of the right eye had developed. The result of computed tomography (CT) was reported as orbital abscess and cellulitis. Although antibiotic treatment was administered for 2 weeks, the exodeviation didn't improve. On ocular examinations performed in our hospital in November-2001, his right eye was esotropic and had a relative afferent pupillary defect. Vision of the right eye was decreased to 0.02. Fundus examination showed optic atrophy. A new CT scan disclosed a foreign body introduced into the right medial orbital wall, nasal cavity and ethmoidal sinus. Although foreign body was surgically removed, vision and eye movement were not improved. In the case of a patient who has undergone orbital trauma, complete history taking and physical examinations must be performed. On suspicion of a foreign body, imaging study such as CT or MRI must be performed. However, because CT findings can be variable, careful follow-up is needed. PMID- 12546449 TI - Corneoscleral cyst treated with distilled water injection. AB - To describe the first case of the treatment of a corneoscleral cyst by distilled water injection into a corneal cyst. The anterior wall of a cyst of the limbal communication was punctured with a surgical blade. Aspiration and irrigation of the contents of the cyst with a 27-gauge anterior chamber cannula were performed repeatedly, three times. Distilled water, instead of balanced salt solution, was injected into the collapsed cyst, and was then aspirated completely after 5 minutes. The injection and aspiration of distilled water was repeated once more. The scleral cyst was surgically excised. Twelve months after surgery, several small white granular opacities, presumably epithelial cell nests, were observed on the interface of the collapsed cyst cavity, but there was no recurrence of the cyst. The best spectacle-corrected visual acuity (BSCVA) was 1.0 with a correction of +1.25-2.00 X 45. No significant change in central corneal endothelial cell density was noted. We suggest that this simple technique may represent an alternative method for the management of corneal cysts, and may have less risk of developing a corneal opacity or causing other serious damage to surrounding tissues. PMID- 12546451 TI - Aspergillosis presenting as an optic neuritis. AB - A 59-year-old woman was referred to our clinic with sudden visual loss in her right eye after she was treated with 40 mg/day of oral prednisolone for 2 weeks under the diagnosis of idiopathic optic neuritis. At that time, computerized tomography (CT) of the brain showed no evidence of optic nerve or brain pathology. However, there was progressive diminution of right visual acuity associated with a limitation of adduction and abduction in the right eye. On magnetic resonance imaging and repeated CT, a malignant lesion was suggested, and was confirmed as an Aspergillus fungus colony by histopathologic examination. Postoperatively, she was treated with intravenous administration of amphotericin B for 13 weeks. However, her condition continued to deteriorate. She developed ptosis and total ophthalmoplegia in the right eye and blindness in both eyes. After discharge, she was given itraconazole for 20 weeks. She has shown no recovery of visual acuity or extraocular motion during a two-year follow-up period. The clinical features of our case suggest that early diagnosis in a case of aspergilloma presenting with visual loss is difficult and that a high index of suspicion, repeated radiological examination and adequate biopsy may be required for diagnosis. PMID- 12546452 TI - Junctional scotoma in giant cerebral aneurysm. AB - A brain lesion located at the lateral side of the sella turcica can produce a junctional scotoma by compressing the ipsilateral optic nerve and the contralateral inferonasal nerve fiber. This study reports a female patient with a junctional scotoma caused by a cerebral aneurysm. At the initial visit, she complained of visual disturbance in both eyes and the right optic disc was atrophied. The visual field showed right blindness and left superotemporal quadrantopsia. A brain CT indicated an approximately 3 cm sized brain mass located superolateral to the sella turcica. The brain MRI showed the lesion to be more like an aneurysm than a pituitary adenoma. Therefore, 4 vessels angiography was done, and this lesion was confirmed to be a sellar variant of an aneurysm located at the right carotid siphon. Like a tumor of the optic chiasm, a cerebral aneurysm can cause visual disturbance and visual field defects. Therefore, an early differential diagnosis is important because the prognosis and treatment of an aneurysm differ. PMID- 12546453 TI - Fast and slow blood-feeding durations of Aedes aegypti mosquitoes in Trinidad. AB - The duration of blood feeding by similar-sized Aedes aegypti (L.) was determined under laboratory conditions in Trinidad, West Indies. Ae. aegypti from the F0 generation contained individuals that fed quickly (106.2 seconds) as well as slowly (330.0 seconds). Among the fast feeders (<2 min), the time taken for blood feeding during the first gonotrophic cycle was much shorter than that observed for the 2nd, 3rd and 4th gonotrophic cycles. In contrast, among the slow feeders the time taken was much slower in the first gonotrophic cycle than that observed in subsequent gonotrophic cycles. Similarly, fast and slow feeding rates did not persist among the progeny of the F0 individuals reared to F1, F2, F3 and F4 generations. The size of the egg clutches was not significantly different for fast- and slow-feeding Ae. aegypti mosquitoes, with mean number of eggs being 85.1 +/- 21.6 for fast feeders and 83.8 +/- 17.7 for slow feeders during the first gonotrophic cycle. Similar patterns were observed among F1, F2, F3 and F4 generations with mean number of eggs being 86.4 +/- 14.8 (range 70-179 eggs) and 87.9 +/- 15.7 (range 68-121 eggs) for the fast and slow feeding females, respectively. These results indicate that mosquitoes have the ability to increase or decrease feeding durations as required, which may counter any physical interference during the blood-feeding process and may consequently enhance vector efficiency. PMID- 12546454 TI - The swarming and mating behaviour of Anopheles gambiae s.s. (Diptera: Culicidae) from Sao Tome Island. AB - The swarming and mating behaviours of the forest cytoform of Anopheles gambiae s.s. were investigated on 194 evenings and 14 mornings between April 1997 and November 1999 in a peri-urban area of the island of Sao Tome, West Africa. Males swarmed 2-3 m above markers of horizontal contrast such as those formed between grass areas and footpaths, or bushes. Evening swarms started 2 min before sunset in sheltered sites and a minute or two later in exposed ones. It took approximately 5 mins from the arrival of the first male for the swarm to reach estimated maximum numbers. Mating pairs were first seen 7 min after the start of swarming. Maximum numbers of pairs in copula were observed 8 min later. Up to 270 pairings were seen in the 20 min period before darkness. Removal of males had no effect on the number of females arriving at the swarm. Males were attracted to sounds that approximated the female flight tone but not to filter paper samples of squashed virgin females swung through the swarm. A much-reduced amount of swarming and a single mating were recorded at dawn. The same locations for swarming were used at different times and at different heights by ants, Culicoides sp. and Culex quinquefasciatus. PMID- 12546455 TI - A review of the North American species of Oropsylla Wagner and Ioff, 1926 (Siphonaptera: Ceratophyllidae: Ceratophyllinae). AB - The North American species of the flea genus Oropsylla are reviewed and their host/parasite relationships and distribution are discussed, as well as their role in the maintenance and transmission of sylvatic plague. Keys to the subgenera and species are presented and illustrations of their diagnostic characteristics are included. PMID- 12546456 TI - Ecology of American cutaneous leishmaniasis in the state of Rio de Janeiro, Brazil. AB - A study of the sand fly fauna was carried out in the endemic area ofAmerican cutaneous leishmaniasis (ACL) in the Municipality of Mesquita, State of Rio de Janeiro, in an area where dwellings (with humans and dogs infected by Leishmania (Viannia) braziliensis) have been recently visited by sloths. Weekly captures were made during two consecutive years in two different sites (1 m and 600 m above sea level) and in three different habitats in each site (domestic, extra domestic and sylvatic) using CDC light traps and Shannon traps. The following species were identified: Brumptomyia (Brumptomyia) guimaraesi, B. (B.) nitzulescui, Lutzomyia (Lutzomyia) longipalpis, L. migonei, L. serrana, L. (Psathyromyia) lutziana, L. barrettoi, L. (Nyssomyia) intermedia, L. (N.) whitmani, L. (Mycropygomyia) schireiberi, L. quinquefer, L. sallesi, L. (Psychodopygus) ayrozai, L. pascalei, L. (Pintomyia) bianchigalatiae and L. lanei. The collection of B. guimaraesi and B. nitzulescui species indicates the proximity of the study area to the sylvatic habitat. Two important suspected vectors of ACL were found: L. intermedia and L. migonei. L. longipalpis, a proven vector of American visceral leishmaniasis (AVL), was also found in this area. L. intermedia, the predominant sand fly species, was collected by both methods of capture and in the three different sites. It was observed biting man inside houses, but it was less anthropophilic in the sylvatic habitat. Our observations suggest that altitude does not influence the dispersion of L. intermedia population in the Municipality of Mesquita. The data indicate that L. intermedia remains as a putative vector of Leishmania (V.) braziliensis in this locality. An hypothesis of a sylvatic transmission cycle in the area and its correlation with the domestic cycle are discussed in the present paper. PMID- 12546457 TI - Effect of deer exclusion by fencing on abundance of Amblyomma americanum (Acari: Ixodidae) on Fire Island, New York, USA. AB - The effects of deer exclusion on northern populations of lone star ticks, Amblyomma americanum, were tested at the Lighthouse Tract, Fire Island, NY, USA, where densities of this species have increased recently. Game fencing was erected to exclude deer from two sites of roughly one ha each, and populations of nymphal and adult A. americanum within were compared with those at control sites outside the exclosures. Percent control of nymphs within vs. outside the exclosures averaged 48.4% in the four years post-treatment, compared to pretreatment values. Percent control varied markedly in different years, suggesting that factors in addition to deer densities had strong effects on population densities of A. americanum. Exclosures of this size did not control adult A. americanum. Effects of deer exclusion in this recently expanded northern population of A. americanum were similar to those that have been reported for southern populations of this species. PMID- 12546458 TI - Selection for pyrethroid resistance in a colony of Anopheles minimus species A, a malaria vector in Thailand. AB - This study tested susceptibilities of Anopheles minimus mosquitoes to deltamethrin during each of 19 generations (although technical problems excluded selective pressure experiments during generations 11-13). The ultimate goal was to establish a pyrethroid resistant colony of this important malaria vector in Thailand. Resistance was selected for by exposing, using the World Health Organization test protocol, sequential generations of An. minimus females to LD50 and LT50 values of deltamethrin. The LD50 and LD90 values were determined for populations from each subsequent generation by probit analysis and significant increases (chi-square test, P>0.01) occurred from one generation to the next. There was approximately a 22-fold increase in the LD50 and a 27-fold increase in LD90 when the F10 generation was compared to the parent colony (F1). Similarly, the LT50 and LT90 values were also increased during selection experiments during generations 14-19. There was roughly a 3-fold increase in susceptibility of F19 females compared to F14 females. In addition, deltamethrin conferred a cross resistance to DDT in the selected colony. Baseline information from these experiments will serve as a guide for future studies on susceptibilities of wild An. minimus populations in Thailand. PMID- 12546459 TI - Field evaluation of cypermethrin and cyfluthrin against dengue vectors in a housing estate in Malaysia. AB - Cynoff 25ULV (cypermethrin 25 g/l) and Solfac UL015 (cyfluthrin 1.5% w/v) were evaluated against the sentinel sugar-fed adults and 4th-instar larvae of Aedes aegypti in a housing estate endemic of dengue in Malaysia. The impact of both pyrethroids on field populations of Aedes albopictus and Aedes aegypti larvae was monitored weekly using bottle containers. Both Cynoff 25ULV and Solfac UL015 showed adulticidal effects and larvicidal effects. This field trial using Cynoff 25ULV against dengue vectors showed its potential for use in dengue vector control programs. PMID- 12546460 TI - A review of the flea genus Eumolpianus Smit, 1983 with a discussion of its geographic distribution and host associations (Siphonaptera: Ceratophyllidae: Ceratophyllinae). AB - The genus Eumolpianus Smit, 1983, a group of fleas infesting chipmunks mostly in western North America, is reviewed. Its host preferences and distribution are discussed and the relationships of the 7 currently recognized species are examined. It is concluded that E. eumolpi americanus (Hubbard, 1950) is a junior synonym of the nominate species. Although not formally synonymized, it is postulated, based on morphology and geographic distribution, that E. eutamiadis (Augustson, 1942) and E. fornacis (Jordan, 1937) are conspecific, as are E. orarius (Johnson, 1961) and E. wallowensis (Hubbard, 1947) with E. cyrturus (Jordan, 1929). The status of E. polumus (Traub & Johnson, 1952) remains uncertain, but it is evidently a member of the E. cyrturus-group. PMID- 12546461 TI - An improved excito-repellency test chamber for mosquito behavioral tests. PMID- 12546462 TI - Selected literature references to vector ecology--April-September 2002. PMID- 12546463 TI - Noninvasive measurement of viscosity from damping of capillary waves. AB - Capillary waves are surface waves on fluids with wavelengths in the millimeter range. The determination of viscosity from the damping of capillary waves is of great practical importance as it affords the possibility of measuring the viscosity of fluids noninvasively. In this paper a noncontact method for generation and detection of capillary waves on fluid is described. A miniature laser interferometer is employed to measure noninvasively the wave amplitude and its attenuation with a resolution of about 10 nm. As a test case, the attenuation data for capillary waves on pure water are used to obtain the kinematic viscosity of water as a function of temperature. The results compare favorably with the most reliable published data on the subject. PMID- 12546464 TI - Sensor data validation using gray models. AB - A new method based on the gray model is described for the online validation of measurements. A gray model is a differential equation describing the behavior of an accumulate generating operation (AGO) data sequence. First-order gray models are fitted to measuring data records using the recursive orthogonal least-squares algorithm. Predictions derived from the fitted gray model are then compared with the actual measurements to generate a prediction error sequence. The quality of the measured value is determined by the prediction errors and variance of the prediction error sequence. Experimental results for detecting the quality of measurements from a thermistor are presented. PMID- 12546465 TI - Feedforward suppression of force ripple based on a simplex-optimized dither signal. AB - This paper presents the design and realization of a feedforward dither signal to reduce the force ripple in an iron-core permanent magnet linear motor (PMLM). A composite control structure is used, consisting of three components: a simple feedforward component, a PID feedback component, and a ripple compensator (RC). The first two components are designed based on a dominant linear model of the motor. The dither signal is generated based on a signal model which is identified using a multidimensional simplex downhill method. In this way, a simple approach is available to eliminate or suppress the inherent force ripple, thus facilitating smooth precise motion while uncompromising on the maximum force achievable. Real-time experimental results verify the effectiveness of the proposed scheme for high precision motion trajectory tracking. PMID- 12546466 TI - A coded approach to system identification using a modified Golay FIR filter. AB - Several techniques are currently in use for system identification, each having its own level of performance. In this paper we present a coded interrogation approach for statistical system identification; it is based around a modified Golay sequence applied to an adaptive finite impulse response (AFIR) filter system. We show that this technique circumvents some of the shortfalls associated with other comparative approaches. The theory of the proposed technique is developed, and the operating principle of a prototype model to verify it is described. Analysis of the results obtained from experiments, performed on a simple first-order linear time invariant (LTI) system, gave an average deviation of +/- 2.94% from the ideal response. PMID- 12546467 TI - Neural networks for process control and optimization: two industrial applications. AB - The two most widely used neural models, multilayer perceptron (MLP) and radial basis function network (RBFN), are presented in the framework of system identification and control. The main steps for building such nonlinear black box models are regressor choice, selection of internal architecture, and parameter estimation. The advantages of neural network models are summarized: universal approximation capabilities, flexibility, and parsimony. Two applications are described in steel industry and water treatment, respectively, the control of alloying process in a hot dipped galvanizing line and the control of a coagulation process in a drinking water treatment plant. These examples highlight the interest of neural techniques, when complex nonlinear phenomena are involved, but the empirical knowledge of control operators can be learned. PMID- 12546468 TI - An approach to improve the offshore platform coordinates accuracy by using multichannel Kalman filtering. AB - In this paper, multichannel Kalman filters for estimation of offshore platform (OP) coordinates are designed. The complete OP motion is assumed to be composed of the low-frequency motion caused by the wind and undercurrent, and the high frequency motion caused by the sea. The mathematical model of the low-frequency OP motion is given by the normal differential equation system, and the high frequency OP motion is represented by a moving-average multivariable autoregression model. The parameter estimation problem for the model of the low frequency OP motion, on which the in-service control is performed, is solved through two jointly operating Kalman filters: the first one is for the estimation of the parameters of the low-frequency motion model, and the second one is for the parameter estimation for the high-frequency model. The parameters of the first filter are automatically adapted to variations of the second filter, i.e., they are adapted to disturbances from the sea. Two algorithms for the OP motion parameter estimation (parallel and with preliminary data compression). employed for several measuring channels data estimation, are developed, and simulated on a computer. Some recommendations on their use are given. PMID- 12546469 TI - Auto-tuning of cascade control systems. AB - In this paper, a novel auto-tuning method for a cascade control system is proposed. By employing a simple relay feedback test, both inner and outer loop model parameters can be simultaneously identified. Consequently, well-established proportional-integral-derivative (PID) tuning rules can be applied to tune both loops. Compared with existing methods, the new method is simpler and yet more effective. It can be directly integrated into commercially available industrial auto-tuning systems. Some examples are given to illustrate the effectiveness and robustness of the proposed method. PMID- 12546470 TI - Digital redesign of anti-wind-up controller for cascaded analog system. AB - The cascaded conventional anti-wind-up (CAW) design method for integral controller is discussed. Then, the prediction-based digital redesign methodology is utilized to find the new pulse amplitude modulated (PAM) digital controller for effective digital control of the analog plant with input saturation constraint. The desired digital controller is determined from existing or pre designed CAW analog controller. The proposed method provides a novel methodology for indirect digital design of a continuous-time unity output-feedback system with a cascaded analog controller as in the case of PID controllers for industrial control processes with the presence of actuator saturations. It enables us to implement an existing or pre-designed cascaded CAW analog controller via a digital controller effectively. PMID- 12546471 TI - Modeling and control of the magnetic suspension system. AB - A fuzzy logic based controller applied to a simple magnetic suspension is presented in this paper. The simple electromagnet-ball system and the contactless optical position measurement system are developed as a physical model of the magnetic suspension. A nonlinear mathematical model is presented and linearized. This model has been used to design a discrete linear PID controller with optimal parameters. The physical real-time model was constructed in order to compare the performance of the linear discrete PID controller and the proposed fuzzy logic based PID controller. The decomposed fuzzy PID controller has proportional, integral, and derivative separate parts which are tuned independently. When testing it becomes clear that the decomposed fuzzy PID controller gives better performance over a typical operational range than a traditional linear PID controller. PMID- 12546472 TI - A new Smith predictor and controller for control of processes with long dead time. AB - Good control of processes with long dead time is often achieved using a Smith predictor configuration. Typically a PI or PID controller is used; however, it is shown in this paper that for some situations improved set point and disturbance responses can be obtained if a PI-PD controller is used. Several methods are possible for selecting the parameters of the PI-PD controller but when the plant transfer function has no zeros, the use of the standard forms provides a simple algebraic approach, and also reveals why difficulties may be encountered if a PID controller is used. Some examples are given to show the value of the approach presented. PMID- 12546473 TI - A PI-PD controller design for control of unstable and integrating processes. AB - A model-based PI-PD controller design, where the PD feedback is used to change the poles of the plant transfer function to more desirable locations for control by a PI controller, is proposed. Several procedures for obtaining the parameters of the PI-PD controller are possible but one of the simplest approaches, which is used in this paper, is to employ integral squared time error standard forms as this enables the design to be completed using simple algebra. Also, an exact method for model extraction of some integrating processes which may or may not have a time delay is presented. The method is compared with several existing methods to control integrating processes and it is shown that the proposed method is superior to existing ones. PMID- 12546474 TI - Optimal operation of batch processes via the tracking of active constraints. AB - This paper presents a new measurement-based optimization framework for batch processes whereby optimal operation can be achieved via the tracking of active constraints. It is shown that, under mild assumptions and to a first-order approximation, tracking the necessary conditions of optimality is equivalent to tracking active constraints (both during the batch and at the end of the batch). Thus the optimal input trajectories can be adjusted using measurements without the use of a model of the process. When only batch-end measurements are available, the proposed method leads itself to an efficient batch-to-batch optimization scheme. The approach is illustrated via the simulation of a semibatch reactor under uncertainty. PMID- 12546475 TI - A method for detection and diagnosis on batch fermentations. AB - In this work we present some basic ideas about detection and diagnosis of faults and abrupt dynamic changes in batch fermentations. Our work focuses on the simultaneous use of two detection methods (residual based and balances based) within the estimation procedure. The idea behind the use of both methods is that the weakness of one of them can be compensated by the use of the other one. Thus the simultaneous use of both methods allows detecting and possibly isolating a wide range of faults. Observations such as the effect of nonlinearities on the detection tests and robustness to model uncertainty are discussed. Numerical results on a particular case, the xanthan gum batch fermentation, are presented. Simulated faults and abnormal behaviors were promptly detected but diagnostics showed mixed results. PMID- 12546476 TI - Practical approach to tuning MPC. AB - This paper presents the results of a heuristic approach for developing model predictive control (MPC) tuning rules. The tuning has been applied and tested in easy-to-use MPC. Process modeling in this MPC uses normalized input/ output range. As a result there is no need for tuning outputs, a procedure known as adjusting equal concern error. Penalties on moves are set as a function of process dead time as the primary factor, with some correction from process gain. The default calculation delivers robust control, which tolerates up to triple increase in process static gain. If control is too aggressive, further on-line adjustment can be done by set point reference trajectory. Test results show that this tuning is robust for process gain change, however, it is much less efficient in compensating for process dead-time changes. It was found that dead-time mismatch is much better compensated with the model correction filter. Combining the three handles, i.e., penalties on moves, reference trajectory, and model filter, easy and intuitively understandable MPC tuning was achieved. The findings are illustrated by numerous MPC simulated tests. PMID- 12546477 TI - Derivation of an equation for quantitative SIL assignment. AB - Quantitative risk analysis for selection of safety integrity levels (SIL's) is becoming more prevalent due to the high equipment costs associated with qualitative methods. Procedures for applying quantitative methods to the SIL selection problem are in the early stages of development. Literature explaining quantitative methods is scarce and many engineers are forced to develop ad hoc procedures, some of which are mathematically flawed. One of the equations popularly used to assign a probability of failure on demand (PFD) to a safety instrumented function (SIF) based on tolerable risk and unmitigated event frequency is shown in the equation below. Variations of this equation are in use by various SIS practitioners: PFD(SIF) = f(Tolerable)_Risk/f(Unmitigated)_Event. This paper presents a rigorous derivation of this equation based on scientific laws of probability math and standardized definitions used in the field of SIS engineering. By publishing this derivation the author hopes to lend more credibility to this type of SIL selection process and increase its usage in the process industries. PMID- 12546478 TI - Developmental change in variability of lip muscle activity during speech. AB - Compared to adults, children's speech production measures sometimes show higher trial-to-trial variability in both kinematic and acoustic analyses. A reasonable hypothesis is that this variability reflects variations in neural drive to muscles as the developing system explores different solutions to achieving vocal tract goals. We investigated that hypothesis in the present study by analyzing EMG waveforms produced across repetitions of a phrase spoken by 7-year-olds, 12 year-olds, and young adults. The EMG waveforms recorded via surface electrodes at upper lip sites were clearly modulated in a consistent manner corresponding to lip closure for the bilabial consonants in the utterance. Thus we were able to analyze the amplitude envelope of the rectified EMG with a phrase-level variability index previously used with kinematic data. Both the 7- and 12-year old children were significantly more variable on repeated productions than the young adults. These results support the idea that children are using varying combinations of muscle activity to achieve phonetic goals. Even at age 12 years, these children were not adult-like in their performance. These and earlier kinematic studies of the oral motor system suggest that children retain their flexibility, employing more degrees of freedom than adults, to dynamically control lip aperture during speech. This strategy is adaptive given the many neurophysiological and biomechanical changes that occur during the transition from adolescence to adulthood. PMID- 12546479 TI - Direct magnitude estimation and interval scaling of naturalness and severity in tracheoesophageal (TE) speakers. AB - The purpose of this study was to determine the psychophysical character and validity of auditory-perceptual ratings of naturalness and overall severity for tracheoesophageal (TE) speech. This was achieved through use of direct magnitude estimation (DME) and equal-appearing interval (EAI) scaling procedures. Twenty adult listeners judged speech naturalness and overall severity from connected speech samples produced by 20 adult male TE speakers. A comparison of DME- and EAI-scaled judgments yielded a metathetic continuum for naturalness and a prothetic continuum for overall severity. These data provide support for the use of either DME or EAI scales in auditory-perceptual ratings of naturalness, but they provide support only for DME scales in judging overall severity for TE speech. The present results suggest that the nature of perceptual phenomena (prothetic vs. metathetic) for TE speakers is consistent with findings for the same dimensions produced by normal laryngeal speakers. These data also support a need for further study of perceptual dimensions associated with TE voice and speech in order to avoid the inappropriate and invalid use of EAI scales frequently found in diagnosis, assessment, and evaluation of this clinical population. PMID- 12546480 TI - Epidemiology of stuttering in the community across the entire life span. AB - A randomized and stratified investigation was conducted into the epidemiology of stuttering in the community across the entire life span. Persons from households in the state of New South Wales, Australia, were asked to participate in a telephone interview. Consenting persons were given a brief introduction to the research, and details were requested concerning the number and age of the persons living in the household at the time of the interview. Interviewees were then given a description of stuttering. Based on this description, they were asked if any person living in their household stuttered (prevalence). If they answered "yes," a number of corroborative questions were asked, and permission was requested to tape over the telephone the speech of the person who stutters. Confirmation of stuttering was based on (a) a positive detection of stuttering from the tape and (b) an affirmative answer to at least one of the corroborative questions supporting the diagnosis. Results showed that the prevalence of stuttering over the whole population was 0.72%, with higher prevalence rates in younger children (1.4-1.44) and lowest rates in adolescence (0.53). Male-to female ratios ranged from 2.3:1 in younger children to 4:1 in adolescence, with a ratio of 2.3:1 across all ages. The household member being interviewed was also asked whether anyone in the household had ever stuttered. If the answer was "yes," the same corroborative questions were asked. These data, along with the prevalence data, provided an estimate of the incidence or risk of stuttering, which was found to range from 2.1% in adults (21-50 years) to 2.8% in younger children (2-5 years) and 3.4% in older children (6-10 years). Implications of these results are discussed. PMID- 12546481 TI - "Pitch" accent in alaryngeal speech. AB - Highly proficient alaryngeal speakers are known to convey prosody successfully. The present study investigated whether alaryngeal speakers not selected on grounds of proficiency were able to convey pitch accent (a pitch accent is realized on the word that is in focus, cf. Bolinger, 1958). The participating speakers (10 tracheoesophageal, 9 esophageal, and 10 laryngeal [control] speakers) produced sentences in which accent was cued by the preceding context. For each utterance, a group of listeners identified which word conveyed accent. All speakers were able to convey accent. Acoustic analyses showed that some alaryngeal speakers had little or no control over fundamental frequency. Contrary to expectation, these speakers did not compensate by using nonmelodic cues, whereas speakers using F0 did use nonmelodic cues. Thus, temporal and intensity cues are concomitant with the use of F0; if F0 is affected, these nonmelodic cues will be as well. A pitch perception experiment confirmed that alaryngeal speakers who had no control over F0 and who did not use nonmelodic cues were nevertheless able to produce pitch movements. Speakers with no control over F0 apparently relied on an alternative pitch system to convey accents and other pitch movements. PMID- 12546482 TI - Articulatory movements in adolescents: evidence for protracted development of speech motor control processes. AB - In order to contribute to a more comprehensive model of speech motor development, we examined the movement trajectories of the upper lip, lower lip, and jaw to determine (a) if there are changes in articulatory motor control in late adolescence; (b) if there are sex differences during this developmental period, perhaps related to differences in craniofacial growth rates; (c) if control of jaw motion is adultlike earlier than control of the upper and lower lip; and (d) if control of spatial and temporal aspects of articulatory movement co-develop in adolescence. Participants were 12-, 14-, and 16-year-olds, and young adults (mean age 21.2 years), with 15 males and 15 females per group. A measure reflecting spatiotemporal consistency in trajectory formation for repeated productions of a phrase was calculated for the upper lip, lower lip, and jaw movements. Overall trajectory variability was higher for adolescents compared to young adults. Jaw trajectories were less variable than upper lip or lower lip trajectories, but all effectors showed parallel decreases in variability as age increased, suggesting that control of jaw movement does not reach adult performance before control of the lips. Separate temporal and spatial measures revealed that adolescents had significantly longer movement durations, lower velocities, smaller displacements, and greater variability on these measures than young adults. There were no sex differences on any measure examined, suggesting that peripheral growth factors do not account for this protracted developmental time course. These results provide initial evidence of significant changes in speech motor control processes during adolescence. PMID- 12546483 TI - Estimating blood PCO2 during speaking: a postscript on Hoit and Lohmeier (2000). PMID- 12546484 TI - A longitudinal investigation of reading outcomes in children with language impairments. AB - This investigation examined the reading outcomes of children with language impairments (LI). A large subsample of children who participated in an epidemiologic study of language impairments in kindergarten (J. B. Tomblin, N. Records, P. Buckwalter, X. Zhang, E. Smith, & M. O'Brien, 1997) was followed into second and fourth grades. Participants' language, reading, and nonverbal cognitive abilities were assessed. Results indicated that children with LI in kindergarten were at a high risk for reading disabilities in second and fourth grades. This risk was higher for children with a nonspecific language impairment (nonverbal and language deficits) than for those with a specific language impairment (deficits in language alone). Children with LI in kindergarten who had improved in spoken language abilities by second and fourth grades had better reading outcomes than those with persistent language impairments. Also, children's literacy knowledge/ experience in kindergarten and their initial reading achievement in second grade were good predictors of subsequent reading outcomes. PMID- 12546485 TI - Effects of prelinguistic milieu teaching and parent responsivity education on dyads involving children with intellectual disabilities. AB - This study tested the effect of a method of facilitating prelinguistic communication on parents' responsivity and children's communication and productive language development. The method involved Responsivity education for the parents and Prelinguistic Milieu Teaching for the children (RPMT). Thirty nine prelinguistic toddlers with intellectual disabilities and their primary caregivers participated in this study. Parent-child pairs were randomly assigned to either the RPMT group or a control group. Communication and language were assessed at study entry and 6, 9, and 12 months later. RPMT facilitated parental responsivity in the posttreatment period. The effect of RPMT on growth rate of child-initiated comments (i.e., the most common type of initiating joint attention) varied by pretreatment measures of that variable. The effect of RPMT on growth rate of child-initiated requests (i.e., the most common type of initiating behavior regulation) varied by presence or absence of Down syndrome. Finally, the effect of RPMT on growth of productive language varied by pretreatment frequency of canonical vocal communication. Recommended alterations in PMT and implications for defining which nonspeaking children are appropriate for prelinguistic goals and treatment were discussed. PMID- 12546486 TI - Discourse comprehension test performance of elders with dementia of the Alzheimer type. AB - Spoken language comprehension, including comprehension for inferential material in narrative discourse, is diminished in dementia of the Alzheimer type (DAT). There are, however, no empirical data concerning comprehension by adults with DAT of main ideas versus details in narratives. Evidence from other groups with and without brain damage has shown that comprehension for main ideas is relatively better than for details and that comprehension for stated material is relatively better than comprehension for inferential material. Participants in the present investigation were 24 older adults, 8 with early-stage DAT (EDAT), 8 with middle stage DAT (MDAT), and 8 with no brain damage (NBD). Selected narratives and associated sets of yes-no questions from the Discourse Comprehension Test (DCT) (Brookshire and Nicholas, 1993) were presented on videotape. Participants with EDAT and MDAT had significantly poorer overall comprehension of DCT narratives than did those in the NBD group (p < .0001), but they did not differ significantly from each other. Responses to DCT narratives by participants in the NBD, EDAT, and MDAT groups followed the same pattern of relatively better comprehension for main ideas than for details and relatively better comprehension for stated than for implied information. Working memory and episodic memory were shown to be significantly associated with DCT overall scores. Together, these findings suggest that although overall narrative comprehension is diminished in those with DAT, individuals appear to retain a mental representation for narratives that facilitates better comprehension of main ideas than of details as well as better comprehension of stated information than of implied information. This interpretation is consistent with schema-based accounts of narrative comprehension. PMID- 12546487 TI - The interaction of ambient frequency and feature complexity in the diphthong errors of children with phonological disorders. AB - This study examined the interaction of ambient frequency and feature complexity in the diphthong errors produced by Cantonese-speaking children with phonological disorders. A total of 611 diphthongs produced by 13 Cantonese-speaking children with speech disorders were subjected to perceptual analysis. The percentage accuracy of production and error patterns was examined. Perceptual analysis showed that /(see text)i/ and /ui/ were most frequently in error, whereas /ei/, /ou/, and /(see text)u/ were least frequently in error. Diphthong errors (usually diphthong reduction) arise as a function of both ambient frequency and feature complexity. The combination of ambient frequency and feature complexity yields a complexity metric reflecting accuracy of production. Treatment guidelines include consideration of three basic factors: ambient frequency, feature complexity, and error patterns. PMID- 12546488 TI - Validity and reliability of the communication and symbolic behavior scales developmental profile with very young children. AB - Three studies were conducted to evaluate the validity and reliability of the three measures of the Communication and Symbolic Behavior Scales Developmental Profile (CSBS DP): (1) a one-page parent-report checklist; (2) a four-page follow up caregiver questionnaire (CQ); and (3) a behavior sample (BS), which is a face to-face evaluation of the child. Participants for these studies were drawn from a pool of 603 children for the checklist and CQ (ages 6-24 months) and 364 children for the BS (ages 12-24 months). Study 1 examined the concurrent relationship of standard scores for the checklist, CQ, and BS. Large correlations were found between the checklist and CQ and moderate to large correlations were found between each of the parent report tools and the BS. Study 2 examined test-retest stability by comparing the raw and standard scores over a 4-month retest interval. The results indicated significantly greater retest raw scores but no significant differences between standard scores from test to retest for the checklist, CQ and BS, providing evidence that the three measures detect growth over short periods but produce relatively stable rankings of children. Study 3 examined the concurrent and predictive relationship of the three CSBS DP measures and children's outcomes on standardized tests of receptive and expressive language at 2 years of age. Moderate to large correlations were found between all of the CSBS DP measures and language outcomes at 2 years of age. Multiple regression analyses indicated that the three composites were a significant predictor of receptive and expressive language outcomes. The findings from these three studies support the use of the CSBS DP as a screening and evaluation tool for identifying children with developmental delays at 12 to 24 months of age. PMID- 12546489 TI - Nominal versus verbal morpheme use in late talkers at ages 3 and 4. AB - Late talkers with normal receptive language and typically developing peers matched at 24- to 31-month intake on socieoeconomic status and nonverbal cognitive skills were compared at age 3 (N = 29, 20) and age 4 (N = 37, 16) on grammatical morpheme suppliance during speech samples. Age 4 late talkers differed from age 3 MLU-matched typically developing children on only the contractible copula. At age 4, "late bloomers" did not differ from typically developing children on any morpheme, but late talkers with "continuing delay" differed from comparison children on articles, nominative case pronouns, auxiliary be, and the contractible copula. Noun phrase morphemes were acquired earlier than verb phrase morphemes by both late talkers and comparison children, a nominal-verbal morpheme "decalage" that was first reported by R. Brown (1973). Results suggested that our late talkers did not have a selective deficit in verb morphology relative to their MLU. Findings are discussed in terms of a spectrum of SLI, with both late talkers and preschoolers with SLI hypothesized to have weaker endowments for language learning than typically developing children, but with late talkers being less impaired and thus closer to normal on this spectrum. PMID- 12546490 TI - Story narratives of adults with closed head injury and non-brain-injured adults: influence of socioeconomic status, elicitation task, and executive functioning. AB - Narratives were elicited in two story tasks, retelling and generation, from two groups of adults, 55 with closed head injury (CHI) and 47 non-brain-injured (NBI), recruited from rehabilitation facilities in three northeastern states. Participants were classified, on the basis of their socioeconomic status (SES), as professional, skilled worker, or unskilled worker. Narratives were analyzed using five discourse measures at the levels of sentence production, intersentential cohesion, and story grammar. Discourse performance was then compared across groups, tasks, and SES levels. Discourse performance of the CHI group was also compared with their scores from the Wisconsin Card Sorting Test, a measure of executive functioning. Results indicated that two discourse measures distinguished the groups. The CHI participants produced significantly fewer words per T-unit and fewer T-units within episode structure than did the NBI group, which was attributed to difficulties with content organization. Performance on all five discourse measures differed for the story retelling versus the story generation tasks for both CHI and NBI groups. All participants produced longer and more grammatically complex T-units in the story generation task than in story retelling. However, cohesive adequacy and story grammar were better in the story retelling task than in the story generation task. It was therefore concluded that story generation was a more challenging task than story retelling for both groups. The only significant difference noted for SES involved the measure of intersentential cohesion. The unskilled workers demonstrated poorer cohesive adequacy than either the skilled workers or professionals, regardless of group or story task. Finally, modest correlations were noted between the discourse performance of the CHI group and scores from the Wisconsin Card Sorting Test in both story tasks. PMID- 12546491 TI - Presbyacusis and the auditory brainstem response. AB - Age-related hearing loss (ARHL or presbyacusis) is an increasingly common form of sensorineural hearing loss (SNHL) as a result of changing demographics, and the auditory brainstem response (ABR) is a common experimental and clinical tool in audiology and neurology. Some of the changes that occur in the aging auditory system may significantly influence the interpretation of the ABR in comparison to the ABRs of younger adults. The approach of this review will be to integrate physiological and histopathological data from human and animal studies to provide a better understanding of the array of age-related changes in the ABR and to determine how age-related changes in the auditory system may influence how the ABR should be interpreted in presbyacusis. Data will be described in terms of thresholds, latencies, and amplitudes, as well as more complex auditory functions such as masking and temporal processing. Included in the review of data will be an attempt to differentiate between age-related effects that may strictly be due to threshold elevation from those that may be due to the aging process. PMID- 12546492 TI - Across-frequency comparison of temporal speech information by listeners with normal and impaired hearing. AB - Listeners with normal hearing (NH) and with sensorineural hearing impairment (HI) were tested on a speech-recognition task requiring across-frequency integration of temporal speech information. Listeners with NH correctly identified a majority of key words in everyday sentences when presented with a synchronous pair of speech-modulated tones at 750 and 3,000 Hz. They could tolerate small amounts (12.5 ms) of across-frequency asynchrony, but performance fell as the delay between bands was increased to 100 ms. Listeners with HI performed more poorly than those with NH when presented with synchronous across-frequency information. Further, performance of listeners with HI fell as a function of asynchrony more steeply than that of their NH counterparts. These results suggest that listeners with HI have particular difficulty comparing and effectively processing temporal speech information at different frequencies. The increased influence of asynchrony indicates that these listeners are especially hindered by slight disruptions in across-frequency information, which implies a less robust comparison mechanism. The results could not be attributed to differences in signal or sensation level, or in listener age, but instead appear to be related to the degree of hearing loss. This across-frequency deficit is unlikely to be attributed to known processing difficulties and may exist in addition to other known disruptions. PMID- 12546493 TI - Influence of hearing loss on the perceptual strategies of children and adults. AB - To accommodate growing vocabularies, young children are thought to modify their perceptual weights as they gain experience with speech and language. The purpose of the present study was to determine whether the perceptual weights of children and adults with hearing loss differ from those of their normal-hearing counterparts. Adults and children with normal hearing and with hearing loss served as participants. Fricative and vowel segments within consonant-vowel consonant stimuli were presented at randomly selected levels under two conditions: unaltered and with the formant transition removed. Overall performance for each group was calculated as a function of segment level. Perceptual weights were also calculated for each group using point-biserial correlation coefficients that relate the level of each segment to performance. Results revealed child-adult differences in overall performance and also revealed an effect of hearing loss. Despite these performance differences, the pattern of perceptual weights was similar across all four groups for most conditions. PMID- 12546494 TI - Temporal integration of sinusoidal increments in the absence of absolute energy cues. AB - Classical temporal integration (TI) is often viewed as a frequency-dependent, energy-based detection process. Detection thresholds for brief sinusoidal increments in either a fixed-level or a random-level broadband pedestal are reported that refute this traditional perspective of TI, Instead, evidence is presented that indicates (a) detection of absolute energy is not necessary for the TI effect and (b) the frequency dependence of TI is consistent with variations across frequency in peripheral auditory tuning, rather than the integration process per se. When peripheral frequency selectivity is controlled, TI can be explained by a frequency-invariant integration process over at least the frequency range from 500 to 4,000 Hz. This process is characterized by threshold improvements of 8-9 dB per decade increase in duration for increment durations between 10 and 300 ms. PMID- 12546495 TI - Spectral contributions to the benefit from spatial separation of speech and noise. AB - Speech recognition in noise improves when speech and noise sources are separated in space. This benefit has two components whose effects are strongest in different frequency regions: (1) interaural level differences (e.g., head shadow), which are largest at higher frequencies, and (2) interaural time differences, which have their greatest contribution at lower frequencies. Binaural interactions enhance the separation of signals from noise through the use of these interaural differences. Here, the benefit attributable to spatial separation was measured as a function of the low- and high-pass cutoff frequency of speech and noise. Listeners were younger adults with normal hearing, older adults with normal hearing, and older adults with hearing loss. Binaural thresholds for narrowband noises were measured in quiet and in a speech-shaped masker as a function of masker low-pass cutoff frequency. Speech levels corresponding to 50% correct recognition of sentences from the Hearing in Noise Test (HINT) were measured in a 65-dB SPL speech-shaped noise. Thresholds for narrowband noises and for speech were measured with two loudspeaker configurations: (1) signals and speech-shaped noise at 0 degrees azimuth (in front of the listener) and (2) signals at 0 degrees azimuth and speech-shaped noise at 90 degrees azimuth (at the listener's side). The criterion measure was spatial separation benefit, or the difference in thresholds for the two conditions. Benefit of spatial separation for unfiltered speech averaged 6.1 dB for younger listeners with normal hearing, 4.9 dB for older listeners with normal hearing, and 2.7 dB for older listeners with hearing loss. Benefit was differentially affected by low-pass and high-pass filtering, suggesting a trade off of the contributions of higher frequency interaural level differences and lower frequency interaural timing cues. As expected, older listeners with hearing loss benefited little from the improved signal-to-noise ratios in the higher frequencies resulting from head shadow, but showed some benefit from lower frequency cues. Spatial benefit for older listeners with normal hearing was reduced relative to benefit for younger listeners. This result may be related to older listeners' elevated thresholds at frequencies above 6.0 kHz. PMID- 12546496 TI - Speckle noise in bar-code scanning systems--power spectral density and SNR. AB - Laser-based flying-spot scanners are strongly affected by speckle that is intrinsic to coherent illumination of diffusing targets. In such systems information is usually extracted by processing the derivative of a photodetector signal that results from collecting over the detector's aperture the scattered light of a laser beam scanning a bar code. Because the scattered light exhibits a time-varying speckle pattern, the signal is corrupted by speckle noise. In this paper we investigate the power spectral density and total noise power of such signals. We also analyze the influence of speckle noise on edge detection and derive estimates for a signal-to-noise ratio when a laser beam scans different sequences of edges. The theory is illustrated by applying the results to Gaussian scanning beams for which we derive closed form expressions. PMID- 12546497 TI - Planar reflection grating lens for compact spectroscopic imaging system. AB - A compact spectroscopic imaging device consisting of a planar reflection grating lens, a probe fiber array, and a two-dimensional image sensor was proposed and discussed. Reflected or luminescent lights from a subject are coupled to the probe fibers, guided to fiber output ends, radiated into the air, diffracted by the grating lens with wavelength-dependent angle, and focused onto lines on the image sensor. Two-dimensional intensity distribution on the image sensor can give one-dimensional spectrum distribution along a specified direction. A grating lens was designed with a fiber array and a CCD image sensor for 100-nm wavelength range and 10-mm fiber array width. A spectral resolution of 5 nm and a spatial resolution of 0.25 mm were experimentally confirmed. PMID- 12546498 TI - Scale and translation invariant minimum average correlation energy filter. AB - A new type, to our knowledge, of scale and translation invariant correlation filter is described. Its form in polar coordinates generalizes the forms of the one-decomposition-term filters. That is combined with the minimum-average correlation-energy optimization method to suppress the sidelobes and achieve correlation peaks. A theoretical analysis as well as a detailed explanation of the computational procedure is provided. The concept is tested on five interferometric images of 256 gray levels without preprocessing. The computed filter gives correlation peaks for reference image scales in the range of 0.3/4.0 (minimal scale: maximal scale = 1:13). The discrimination ability of the filter is investigated-no false peak occurred. The filter works in the background so that there is no need for input image segmentation. A method for extension is described to calculate a filter for more than one reference image. PMID- 12546499 TI - Recovery of blurred images obtained through turbulent media. AB - Method for recovering blurred images taken through turbulent media, such as the atmosphere is presented. In this method amplitude and phase of the object's image are separately determined by special techniques of data processing, and finally the original image can be recovered. Principle and algorithm are described and the techniques to determine amplitude and phase are introduced by use of the results of computer simulations as well as light propagation experiments. As a demonstration to verify the utility of this method, images recovered by this method by use of data taken through a telescope of 1.5 m diameter, and the results of a computer simulation with atmospheric turbulence are shown. The results suggest that the presented method is well suited for the retrieval of blurred images. PMID- 12546500 TI - Digital algorithm for dispersion correction in optical coherence tomography for homogeneous and stratified media. AB - The resolution of optical coherence tomography (OCT) often suffers from blurring caused by material dispersion. We present a numerical algorithm for computationally correcting the effect of material dispersion on OCT reflectance data for homogeneous and stratified media. This is experimentally demonstrated by correcting the image of a polydimethyl siloxane microfludic structure and of glass slides. The algorithm can be implemented using the fast Fourier transform. With broad spectral bandwidths and highly dispersive media or thick objects, dispersion correction becomes increasingly important. PMID- 12546501 TI - Visualization of phase objects by use of a differentiation filter. AB - For the visualization of phase objects by use of a differentiation filter, the phase variation is changed into the intensity variation by differentiation, and then the differentiated image is integrated. In the method used in practice, the differentiated image has been recorded on a film, and then integrated by use of a filter. In this paper, however, the image differentiated by means of a filter is entered into a computer using a CCD camera and then integrated. As a result, the method provides rapid on-line processing. We have performed computer simulations and a detailed analysis of the differentiation filter and also provide the experimental results of three-dimensional image visualization. PMID- 12546502 TI - Resolution improvement with dispersion manipulation and a retrieval algorithm in optical coherence tomography. AB - We propose and demonstrate what is to our knowledge a novel technique of improving the spatial resolution of an optical coherence tomography (OCT) system given a non-Gaussian light source spectrum. By using dispersive materials in the reference arm of the OCT system, the resultant dispersion variation led to a full width at half maximum (FWHM) of the interference fringe envelope smaller than the Fourier transform-limited value of a Gaussian spectral shape with the same spectral FWHM, at the expense of significant tails. The effects of the tails, which would blur the OCT images, were tremendously reduced with a retrieval algorithm. Simulation results and processed OCT scanning images have shown the capability of the proposed technique. PMID- 12546503 TI - Single beam two-views holographic particle image velocimetry. AB - Holographic particle image velocimetry (HPIV) is presently the only method that can measure at high resolution all three components of the velocity in a finite volume. In systems that are based on recording one hologram, velocity components parallel to the hologram can be measured throughout the sample volume, but elongation of the particle traces in the depth direction severely limits the accuracy of the velocity component that is perpendicular to the hologram. Previous studies overcame this limitation by simultaneously recording two orthogonal holograms, which inherently required four windows and two recording systems. This paper introduces a technique that maintains the advantages of recording two orthogonal views, but requires only one window and one recording system. Furthermore, it enables a quadruple increase in the spatial resolution. This method is based on placing a mirror in the test section that reflects the object beam at an angle of 45 degrees. Particles located in the volume in which the incident and reflected beams from the mirror overlap are illuminated twice in perpendicular directions. Both views are recorded on the same hologram. Off-axis holography with conjugate reconstruction and high-pass filtering is used for recording and analyzing the holograms. Calibration tests show that two views reduce the uncertainty in the three-dimensional (3-D) coordinates of the particle centroids to within a few microns. The velocity is still determined plane-by plane by use of two-dimensional particle image velocimetry procedures, but the images are filtered to trim the elongated traces based on the 3-D location of the particle. Consequently, the spatial resolution is quadrupled. Sample data containing more than 200 particles/mm3 are used for calculating the 3-D velocity distributions with interrogation volumes of 220 x 154 x 250 microm, and vector spacing of 110 x 77 x 250 microm. Uncertainty in velocity is addressed by examining how well the data satisfies the continuity equation. The results show significant improvements compared with previous procedures. Limitations of the technique are also discussed. PMID- 12546504 TI - Optimal re-referencing rate for in-plane dynamic speckle interferometry. AB - We investigate experimentally the optimal rate at which the reference speckle pattern should be updated when dynamic speckle interferometry is used to measure transient in-plane displacement fields. Images are captured with a high-speed camera and phase shifting and phase unwrapping are done temporally. For a wide range of in-plane velocities, up to a maximum of 40% of the Nyquist limit, the random errors in the calculated displacement field are minimized by updating the reference speckle pattern after a speckle displacement of 1/10 of the pixel spacing. The technique is applied to measurements of microscale deformation fields within an adhesive joint in a carbon-fiber epoxy composite. PMID- 12546505 TI - Albumen as a relief recording media for spatial distributions of infrared radiation. Fabrication of interference gratings and microlenses. AB - It is shown that a relief is generated when spatial distributions of infrared light (lambda = 10.6 microm) are recorded on albumen films. The relief can be applied to the fabrication of microelements, such as diffraction gratings and microlenses. Examples are shown. PMID- 12546506 TI - Dynamic demonstration of diffractive optic analog-to-digital converter scheme. AB - Dynamic behavior of an analog-to-digital converter (ADC) based on diffractive optical element(s) (DOE)(s) was studied and found to be in agreement with predictions. The analog signal was translated to an angular deflection of a laser beam by means of an acousto-optic (AO) cell. The number of bits in this experimental demonstration was three, using an eight-element DOE array. The maximum sample rate was found to be 2.5 MS/s, the limiting factor being the transit time for the acoustic wave across the width of the laser beam in the AO cell. The study is intended as a first dynamic demonstration of a proposed ADC scheme previously demonstrated in a quasi-static version. The full potential of the ADC scheme will require the use of a fast tunable diode laser to replace the AO deflection scheme used here. PMID- 12546507 TI - Optimized design of temperature-insensitive optical waveguide coupler with 120-nm bandwidth using fluorinated polyimide. AB - Method for designing optimized temperature-insensitive optical waveguide couplers by use of fluorinated polyimide is presented. Based on measured temperature and dispersion characteristics of fluorinated polyimide, a 3-dB waveguide coupler with a 120-nm bandwidth with minimal temperature variance is designed and verified through simulation based on three-dimensional beam propagation. The coupling ratio of the theoretical device is 50 +/- 0.7% in the waveband 1490 to approximately 1610 nm and the temperature range -10 to approximately 40 degrees C. PMID- 12546508 TI - Fixed-length two-dimensional modulation coding for imaging page-oriented optical data storage systems. AB - We present a comprehensive discussion of modulation coding for page-oriented optical data storage (PODS) systems that write and read data in a two-dimensional (2-D) bit image format. We give several 2-D mathematical models for these systems, including two-photon optical data storage systems. Using these models, we describe the nature of intersymbol interference (ISI) in imaging PODS systems and find that its characteristics are different from ISI in conventional serial magnetic and optical data storage systems. To overcome the ISI in these imaging PODS systems, we present what is, to our knowledge, a novel 2-D modulation coding scheme. We also present many examples of fixed-length 2-D modulation codes with diverse properties. Finally, we analyze and compare the bit-error rate performance of these codes. PMID- 12546509 TI - Endoscopic mucosal resection: a promise or an established reality? PMID- 12546510 TI - Does length really matter? ... Oral and topical 5-aminosalicylic acid therapy for ulcerative colitis. PMID- 12546511 TI - Autonomic nerve dysfunction in pathologically obese patients. AB - BACKGROUND: Obese patients frequently present clinical symptoms related to gastrointestinal motility alterations and autonomic nervous system dysfunction. AIM: To evaluate the possible correlation between cardiovascular autonomic nervous dysfunction and oesophageal motility in pathologically obese patients. PATIENTS AND METHODS: Enrolled in the study were 22 patients with a body mass index of 45.72 +/- 7.48 and 10 control subjects, all within 20% of their ideal weight. Oesophageal motility was measured by stationary manometry and scintigraphic transit. Tests for the evaluation of autonomic nervous system were: Valsalva ratio, deep breathing, sustained handgrip, sudormotor axon reflex test and spectral analysis of the variability of R-R interval. RESULTS: The mean pressure of oesophageal peristaltic waves in patients and controls was 39.36 +/- 14 mmHg and 73 +/- 12 mmHg, respectively The scintigraphic mean transit time was 22.96 +/- 16.26 seconds in patients and 10.23 +/- 16.26 seconds in controls (p < 0.001). Spectral analysis of the variability of the R-R interval showed an increase in the parasympathetic component both in the lying and standing position compared to controls. The other autonomic nervous system function tests showed no significant difference between obese patients and controls. CONCLUSIONS: These results suggest that obese patients present a reduction of oesophageal transit and autonomic nervous system dysfunction albeit no direct correlation was found between these phenomena. PMID- 12546512 TI - Autonomic imbalance during the day in patients with inflammatory bowel disease in remission. Evidence from spectral analysis of heart rate variability over 24 hours. AB - BACKGROUND: Autonomic function in inflammatory bowel disease has not yet been studied by means of analysis of 24-hour heart rate variability. AIM: To measure heart rate variability in inflammatory bowel disease patients in remission. PATIENTS AND METHODS: Study population comprised 27 patients with inflammatory bowel disease in remission and 28 healthy, sex- and age-matched controls. Two frequency ranges were analysed: low frequency (0.06-0.15 Hz) and high frequency (0.15-0.40 Hz). RESULTS: Mean values of low frequency and low frequency/high frequency ratio were lower in patients than in controls (p < 0.001). High frequency in patients tended to be higher than in controls (p = 0.09). The only factor that had a marginal effect on heart rate variability indexes was age. In high frequency, there was a significant time effect (p = 0.001) for both groups. There was also a significant time effect in low frequency/high frequency ratio in both groups (p < 0.001). During daytime, the mean values in low frequency/high frequency ratio were lower in patients than in controls (p < 0.001). CONCLUSIONS: There is a shift in the autonomic balance in patients with inflammatory bowel disease in remission towards a condition of relative parasympathetic predominance, which, in the first place, reflects a sympathetic pullback. This imbalance has a circadian rhythm and it is more pronounced during the day. PMID- 12546513 TI - Frequency of extraintestinal manifestations in patients with inflammatory bowel disease in Northwest Greece and review of the literature. AB - BACKGROUND: Inflammatory bowel disease mainly affects the bowel but also has extraintestinal manifestations. AIMS: To report the frequency of extraintestinal manifestations in patients with inflammatory bowel disease in Northwest Greece. PATIENTS; The data files of 256 inflammatory bowel disease patients (diagnosis between 1983-19971 were analysed. METHODS: Retrospective investigation of patient files. RESULTS: In patients with ulcerative colitis: 13.9% (30/215) had developed skin manifestations, 6% (13/215) had kidney stones, 1.39% (3/215) had iridocyclitis, 1.86% (4/215) had primary sclerosing cholangitis, 4.18% (9/215) had sacroiliitis, 8.31% (18/215) had peripheral arthalgias, 2.3% (5/215) had colitic arthritis and finally 1.39% (3/215) had deep vein thrombosis). In patients with Crohn's disease: 24.3% (9/37) had developed skin manifestations, 5.4% (2/37) had kidney stones, 2.7% (1/37) had iridocyclitis, 16.2% (6/37) had sacroiliitis, 8.1% (3/37) had peripheral arthralgias, 5.4% (2/37) had colitic arthritis and, finally, 8.1% (3/37) had deep vein thrombosis. Sacroiliitis (p = 0.01), deep vein thrombosis (p = 0.04) and erythmema nodosum (p = 0.01) were more common in patients with Crohn's disease. CONCLUSIONS: Extraintestinal manifestations are not rare in patients with inflammatory bowel disease, especially in Crohn's disease patients, in our area, but have, generally, a mild profile. PMID- 12546514 TI - Oral and topical 5-aminosalicylic acid (mesalazine) in inducing and maintaining remission in mild-moderate relapse of ulcerative colitis: one-year randomised multicentre trial. AB - BACKGROUND: The association of oral 5-aminosalicylic acid (mesalazine) and enema is effective in treatment of mild-moderate forms of ulcerative colitis. However no study has been aimed at determining optimal duration of this association in active ulcerative colitis. AIM: To determine whether longer duration of therapy: 1. increases the rate of patients achieving remission, and 2. reduces relapse rate during the maintenance period in patients in remission. PATIENTS AND METHODS: A total of 149 patients, (89 male, 60 female), were randomly assigned to a regimen with 5-aminosalicylic acid tablets 2.4 g/day associated with 5 aminosalycilic enema 2 g/day for a 4-week (n = 73) or 8-week regimen (n = 76). After this acute therapy, patients were submitted to clinical, endoscopic and histological examinations and those in remission were assigned to a follow-up (maintenance) period with oral mesalazine alone at a dosage of 1.2 g/day. A clinical visit, including laboratory tests, at 6 months and an endoscopic histological control at 12 months were carried out to exclude symptoms and endoscopic-histological signs of activity. Relapse of disease, i.e., presence of clinical symptoms or abnormal laboratory tests, was confirmed by endoscopy and histology. RESULTS: At end of acute phase, clinical, endoscopic and histological remission was comparable in the two groups: 42/76 (55%), in the 4-week, and 47/73 patients (64%), in the 8-week regimen. No difference was found stratifying patients according to extension of disease. Of these 89 patients in remission, 75 (34 from 4-week regimen; 41 from 8-week regimen) completed 12 months' follow-up. At end of follow-up, a similar percentage of patients in the 4-week regimen (50%) and 8-week regimen (51%) were still in remission. No significant difference between cumulative relapse rates of the two groups was found. Stratifying patients according to extension of disease, in the 8-week regimen group, those with left-sided colitis showed a higher remission rate than that of patients with diffuse colitis (66% versus 35%, p < 0.05). All regimens were well tolerated by most patients during the entire study period. CONCLUSIONS: An additional 4 weeks of topical treatment does not increase the remission rate in patients with mild moderate active ulcerative colitis but seems to reduce the probability of relapse in patients with left-sided colitis. PMID- 12546516 TI - Hepatitis C virus RNA quantitation in hepatic veins and peripheral blood in patients with liver cirrhosis: evidence for low level intrahepatic hepatitis C virus replication in advanced liver disease. AB - BACKGROUND: Very few data exist concerning the level of hepatitis C virus replication within the cirrhotic liver and its relationship to disease severity and progression. AIMS: To quantitate hepatitis C virus RNA in hepatic vein blood and peripheral blood in patients with cirrhosis, to evaluate the correlation of hepatitis C virus levels in paired blood samples, and to compare the results with clinical features. PATIENTS: A series of 25 patients with hepatitis C virus related liver cirrhosis undergoing hepatic vein catheterization were studied: 11 belonged to Child Pugh class A, 8 to class B and 6 to class C. RESULTS: Hepatitis C virus RNA levels did not differ between hepatic vein blood and peripheral blood (p = 0.26), despite a trend towards higher peripheral hepatitis C virus RNA levels. Hepatitis C virus RNA levels did not differ between patients with genotype 1b and non-1b either in hepatic veins or peripheral blood. Hepatitis C virus loads varied according to the severity of cirrhosis. The patients with more severe liver disease had significantly lower RNA titres than those with less advanced cirrhosis, both in hepatic veins (p = 0.002) and peripheral blood (p = 0.004). No differences in hepatitis C virus load were observed between patients in Child Pugh classes B and C. CONCLUSIONS: The present data show that in patients with cirrhosis hepatitis C virus RNA concentrations do not differ between hepatic blood and peripheral blood and, furthermore, confirm that hepatitis C virus replication is reduced in patients with advanced cirrhosis, compared with patients with less severe liver disease. These findings might indicate that patients with liver cirrhosis maintain an efficient intrahepatic hepatitis C virus replication even in end-stage disease, although hepatitis C virus viraemia decreases according to the severity of liver disease. PMID- 12546515 TI - Cytokine, infiltrating macrophage and T cell-mediated response to development of primary and secondary human liver cancer. AB - BACKGROUND: Kupffer cells, monocytes and infiltrating T cells have been considered the major source of interleukin-1beta and tumour necrosis factor-alpha in the liver. AIMS; To explore the expression of interleukin-1beta and tumour necrosis factor-alpha and to evaluate the density and the distribution of T lymphocytes and monocytes/macrophages in the liver of patients with primary and secondary tumours. METHODS: Tumoural and peritumoural liver samples were examined from 21 patients with hepatocellular carcinoma, 10 with hepatic metastases, 5 with benign focal liver lesions and 4 healthy adult livers. Interleukin-1beta and tumour necrosis factor-alpha mRNAs were detected by a semiquantitative comparative reverse transcriptase polymerase chain reaction. T lymphocytes and monocytes/macrophages were detected by immunohistochemistry. RESULTS: Higher levels of interleukin-1beta, tumour necrosis factor-alpha, CD3+ and CD68+ cells were found in the tissue surrounding hepatocellular carcinoma and metastases than in the tumour itself. A strong expression of CD68+ and CD3+ cells was found mainly along the tumour-host interface but the highest expression of CD3+ cells was found at the metastasis interfaces. Interleukin-1beta expression, CD3+ and CD68+ cell densities were higher in peritumoural samples than in so-called "normal" liver tissue. CONCLUSIONS: An increased production of interleukin-1beta and, to a lesser extent, of tumour necrosis factor-alpha mRNA coincides with the presence of cancer be it primary or secondary, both in healthy and cirrhotic livers. The presence of cancer, irrespective of the presence of underlying liver damage, appears to play the most important role. PMID- 12546517 TI - Endoscopic resection of a cavernous haemangioma of the stomach. AB - A lesion, suspected, from the endoscopic appearance, to be an isolated gastric haemangioma was incidentally detected in a 66-year-old male. At follow-up endoscopy, 5 years later, tumour size had increased and its appearance had changed with respect to previous examinations. On endoscopic ultrasonography, the tumour was solid and confined mainly to the submucosal layer of the gastric wall. Examination of biopsy specimens failed to provide any useful information. After biopsy, tumour size and the area of reddish discoloration decreased and endoscopic ultrasonography revealed multiple small cystic lesions in the tumour. Endoscopic resection was then performed with complete excision of the tumour without complications or recurrence. Pathological examination of resected tissue demonstrated cavernous haemangioma of the stomach. PMID- 12546518 TI - Transmission of viral hepatitis by blood and blood derivatives: current risks, past heritage. AB - For more than 40 years in the history of transfusion medicine, transmission of viral hepatitis from infected donors to recipients has been a frequent and serious adverse effect of the administration of blood components and plasma derivatives. This epidemic is now over, at least in developed and resource-rich countries. Hence, the attention of clinicians and investigators now focuses mainly on the measures to reduce the residual risk, on the possible emergence of novel or undiscovered agents causing post-transfusion hepatitis, and on the long term outcome of patients who became infected more than ten years ago. The present article reviews these issues. PMID- 12546519 TI - Duodenal gastric metaplasia, duodenal ulcer and Helicobacter pylori infection. PMID- 12546520 TI - Five cases of nodular gastritis and gastric cancer: a possible association between nodular gastritis and gastric cancer. PMID- 12546521 TI - It is now time for a powerful view in the management of lung cancer. PMID- 12546522 TI - An update on management of lung cancer--Acta Oncologica Lecture-Vasteras 21 March 2002. AB - Globally, the incidence of lung cancer continues to rise drastically in parallel with increased tobacco consumption but with wide geographic variations. Despite the enforcement of more rigid health policies in many countries, lung cancer will for many years to come be a recurrent disease which most physicians will be confronted with at regular intervals. Furthermore, variations in the management of lung cancer occur from region to region influenced by differences in traditions, knowledge, healthcare systems and available resources. In this article a brief overview is presented of the management of both non-small cell and small cell lung cancer including recent results that may have an impact on the management of this disease. PMID- 12546523 TI - Focus on the individual--quality of life assessments in oncology. AB - In this review we investigate how assessments of quality of life (QoL) can be used in daily clinical practice. The focus is on individualized QoL assessments, but computerized-adaptive testing and interpretation of QoL scores are also considered. Quality of life (QoL) measurements have been used relatively infrequently in clinical practice, although individualized care planning and follow-up based on QoL information may lead to better outcomes of treatment and informed and autonomous decision-making by patients. A recent approach in assessing QoL is by individualized measures, which tap QoL as defined by the individual patient. Individualized QoL instruments, such as the Schedule for the Evaluation of Individual Quality of Life (SEIQoL) and the Patient-Generated Index (PGI), have obvious appeal for use in clinical practice, since they incorporate topics of greatest concern to the individual patient, while also capturing their ratings and weightings. However, before general use in daily clinical practice can be recommended, some aspects of these instruments require further research, particularly issues concerning reliability, feasibility, interpretation and effectiveness. PMID- 12546524 TI - Erythropoietin treatment in metastatic breast cancer--effects on Hb, quality of life and need for transfusion. AB - Erythropoietin is an effective treatment for anemia in patients with various types of cancers, but few studies have evaluated the benefit of treatment in advanced breast cancer. In this multicenter study, we investigated the influence of two different doses of epoetin-beta on the level of hemoglobin, the need for blood transfusion, quality of life and safety aspects in patients with metastatic breast cancer. A total of 180 patients were randomized to receive either 1000 IE or 5000 IE epoetin-beta subcutaneously three times per week for 24 weeks. An increase of 20 g/L was defined as a positive hemoglobin response. Blood transfusions were given, if clinically indicated. Additional laboratory values and adverse events were recorded. Quality of life was measured with the aid of the EORTC QLQ-C30 questionnaire. Hemoglobin levels increased significantly in both groups. In the high-dose group, the initial mean Hb value was 98 g/L (64 110), which increased to 121 g/L (83-165) by week 24. In the low-dose group, the mean Hb value was 99 g/L (77-110.5) and by week 24 it was 116 g/L (81-144). The majority of patients who responded to treatment did so during the first four weeks. After 4 weeks, 7 patients in the low-dose group and 24 patients in the high-dose group had increased their Hb values by more than 20 g/L. The need for transfusion was low and did not differ between the groups. Quality of life was significantly enhanced in both groups, and there was no difference in the global quality of life between the two study arms. Epoetin-beta is a well-tolerated, safe and effective treatment of anemia in patients with metastatic breast cancer. There were significant improvements in Hb levels and quality of life in both groups. PMID- 12546525 TI - Dose-effect relationship of bolus 5-fluorouracil in the treatment of advanced colorectal cancer. AB - The effect of different dose intensities of 5-fluorouracil (5-FU) in advanced colorectal cancer was investigated. A total of 312 patients were randomized to receive 400 mg/m2 (group A), 500 mg/m2 (group B) or 600 mg/m2 (group C) of 5-FU with leucovorin 60 mg/m2 on two consecutive days every second week. Treatment continued to progression. Pharmacokinetic analyses with calculation of the area under the concentration (AUC) were performed in 91 patients. The primary endpoint was survival, and secondary endpoints were time to disease progression, toxicity and, if the disease was measurable, tumour response. The study was well balanced in the three groups with respect to a number of patient characteristics. Crude survival as estimated by Kaplan-Meier plots was not statistically significantly different (p = 0.07) but tended to show the best results in the intermediate dose group (median survival 10, 12.5 and 10 months, respectively). Analyses of time to progression or death showed significant differences among the three groups (p = 0.02) with the longest progression-free interval in the intermediate group receiving 500 mg/m2. The objective response rates were 23%, 39% and 28%, respectively (p = 0.02). The actual/projected dose intensity (mg/m2/week) was 92%, 92% and 84%, respectively. AUC did not correlate with response or survival. The frequency of severe side effects in group C was significantly higher than that of groups A and B. The study indicated that an increase from 800 to 1000 mg/m2 of bolus 5-FU fortnightly improved the treatment results but a further increase only worsened the toxicity. PMID- 12546526 TI - Long-term symptoms after external beam radiation therapy for prostate cancer with three or four fields. AB - The aim of this study was to investigate whether external beam radiation treatment with three or four fields affects the risk of long-term distressful symptoms. The study included 145 patients who had been treated in Stockholm from 1993 to 1996 for localized prostate cancer. Bowel, urinary and sexual function as well as symptom-induced distress were assessed by means of a postal questionnaire 29-59 months after therapy. Among patients treated with a multileaf collimator, defecation urgency, diarrhoea and loose stools were more common after four fields than after three fields, but faecal leakage necessitating the use of pads and distress from the gastrointestinal tract were less common (although not statistically significantly so). Among bowel symptoms, the strongest association with gastrointestinal distress was found for faecal leakage. Three fields without a multileaf collimator entailed a higher risk of defecation urgency than three fields with a multileaf collimator. We conclude that the choice of three or four fields may imply a contrasting risk scenario for defecation urgency or diarrhoea in comparison with faecal leakage. PMID- 12546527 TI - Changes in the colonic enteric nervous system in rats with chemically induced colon dysplasia and carcinoma. AB - The enteric nerve plexus in the colon was investigated in rats with chemically induced colonic adenocarcinoma. Tissue specimens from the colons of four group rats, namely controls, treated animals without development of colonic macro- or microscopic changes, rats with dysplasia and lymphoid hyperplasia, and rats with colonic adenocarcinoma were studied using immunocytochemistry, and quantified by computerized image analysis. No morphometeric changes were found in the treated rats regarding the myenteric and submucosal ganglia, with the exception of nitric oxide synthase (NOS), where the number of nerve cell bodies/ganglia was reduced in the myenteric ganglia in rats with both lymphoid hyperplasia and dysplasia, and carcinoma. The relative volume density of protein gene product (PGP) 9.5 immunoreactive (IR) nerve fibres was higher in the muscularis propria in rats with lymphoid hyperplasia and dysplasia, and carcinoma. However the relative volume density of PGP 9.5-IR nerve fibres was higher in the submucosa in rats with carcinoma only. The relative volume density of substance P- and VIP-IR nerve fibres was significantly higher in the muscularis propria in rats with colonic carcinoma. The relative volume density of NOS-IR nerve fibres was significantly decreased in both muscularis propria and submucosa in rats with lymphoid hyperplasia and dysplasia, and carcinoma. These findings imply that regulatory signals of the enteric innervation may be involved in the pathogenesis of colorectal cancer. PMID- 12546528 TI - Overexpression but lack of mutation and methylation of p73 in hepatocellular carcinoma. AB - p73 is one of the family proteins that share structural and functional homologies with the tumor suppressor p53. To analyze the status of p73 in hepatocellular carcinoma (HCC), the allelic loss, allelic expression, mutation and methylation status of the p73 gene were examined in 18 paired HCC and normal tissues. No allelic loss was found. All heterozygous individuals contained RNA of both alleles, indicating that p73 was biallelically expressed in the liver. Notably, semiquantitative reverse transcriptase polymerase chain reaction analysis showed that p73 was consistently overexpressed in the cancerous tissues. Single-stranded conformation polymorphism and sequencing analysis revealed several polymorphisms, but no mutations were found in the entire coding sequence. Finally, the methylation patterns in the promoter and exon 1 regions of p73 were not altered in the cancerous tissues. These results do not support p73 as a tumor suppressor in HCC, but suggest that overexpression of p73 may in some way be associated with the pathogenesis of HCC. PMID- 12546529 TI - Risk factors for breast cancer in Turkish women with early pregnancies and long lasting lactation--a case-control study. AB - A hospital-based case-control study was carried out among 504 women with breast cancer and 610 controls to analyse the risk factors for breast cancer in Turkey. Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for each risk factor were obtained from logistic regression analysis. Risk factors for breast cancer were found to be long-term lactation (> or = 5 years versus never OR 0.31, 95% CI 0.12-0.79), young age at menarche (< 15 years versus > or = 15 OR 1.72, 95% CI 1.30-2.28), late age at first full-term pregnancy (> or = 30 versus < 20 OR 2.86, 95% CI 1.32-6.21), oral contraceptive use (ever versus never OR 1.51, 95% CI 1.10-2.08), positive family history (positive versus negative OR 2.81, 95% CI 1.35-5.82), and menstrual irregularity (yes versus no OR 1.61, 95% CI 1.05 2.49). The results of the present study will lead to a better understanding of the risk factors for breast cancer in a developing country. PMID- 12546530 TI - Norwegian oncologists' expectations of intensity-modulated radiotherapy. AB - AIthough intensity-modulated radiotherapy (IMRT) may increase the therapeutic ratio of radiotherapy for a range of malignancies, only a few IMRT treatments have yet been performed in the Nordic countries. The scores derived from a national survey to assess Norwegian oncologists' expectations of IMRT are presented. A questionnaire was distributed to all consultants in oncology at Norwegian radiotherapy clinics. Summary scores of daily general radiotherapy workload (DGRTW), acquaintance with IMRT (AI) and expectations of IMRT (EI) were derived. Thirty-nine questionnaires (67%) were returned from a total of 58 oncologists. The oncologists' scores on the AI scale (mean score: 7.5 out of 21) were rather low. Their AI scores were found to be positively correlated with their DGRTW. Higher scores on the EI scale were documented (mean score: 6.2 out of 14): 15 oncologists (39%) rated IMRT as one of the three major contributors to potentially increased cancer survival. Oncologists treating patients with prostate, head and neck, gastrointestinal and CNS tumours had higher EI scores than the other oncologists (7.7 vs. 5.1; p = 0.01). The Norwegian radiation oncologists' expectations of IMRT are high in terms of both the potential clinical benefit and the rate of implementation. This should encourage the radiotherapy communities to continue (or rapidly initiate) their efforts in providing the routines required for safe implementation of IMRT. PMID- 12546531 TI - Pagetic bone lesions in a patient with early breast cancer--a pitfall for diagnostic misinterpretation. PMID- 12546532 TI - Variable neighborhood search for extremal graphs. 6. Analyzing bounds for the connectivity index. AB - Recently, Araujo and De la Pena gave bounds for the connectivity index of chemical trees as a function of this index for general trees and the ramification index of trees. They also gave bounds for the connectivity index of chemical graphs as a function of this index for maximal subgraphs which are trees and the cyclomatic number of the graphs. The ramification index of a tree is first shown to be equal to the number of pending vertices minus 2. Then, in view of extremal graphs obtained with the system AutoGraphiX, all bounds of Araujo and De la Pena are improved, yielding tight bounds, and in one case corrected. Moreover, chemical trees of a given order and a number of pending vertices with minimum and with maximum connectivity index are characterized. PMID- 12546533 TI - Windows-based guided data capture software for mass-scale thermophysical and thermochemical property data collection. AB - Guided data capture software (GDC) is described for mass-scale abstraction from the literature of experimental thermophysical and thermochemical property data for organic chemical systems involving one, two, and three components, chemical reactions, and chemical equilibria. Property values are captured with a strictly hierarchical system based upon rigorous application of the thermodynamic constraints of the Gibbs phase rule with full traceability to source documents. Key features of the program and its adherence to scientific principles are described with particular emphasis on data-quality issues, both in terms of data accuracy and database integrity. PMID- 12546534 TI - An efficient implementation of a drug candidate database. AB - The recent advances in laboratory technologies have resulted in a wealth of chemical and biological data. The rapid proliferation of a vast amount of data has led to a set of cheminformatics and bioinformatics applications that manipulate dynamic, heterogeneous, and massive data. An example of such application in the pharmaceutical industry is the computational process involved in the early discovery of lead drug candidates for a given target disease. In this paper, an efficient implementation of a drug candidate database is presented and evaluated. This study shows that high performance data access can be achieved through proper choices of data representation, database schema design, and parallel processing techniques. PMID- 12546535 TI - Application of chemical graph theory for automated mechanism generation. AB - We present an application of the chemical graph theory approach for generating elementary reactions of complex systems. Molecular species are naturally represented by graphs, which are identified by their vertices and edges where vertices are atom types and edges are bonds. The mechanism is generated using a set of reaction patterns (sub-graphs). These subgraphs are the internal representations for a given class of reaction thus allowing for the possibility of eliminating unimportant product species a priori. Furthermore, each molecule is canonically represented by a set of topological indices (Connectivity Index, Balaban Index, Schulz TI Index, WID Index, etc.) and thus eliminates the probability for regenerating the same species twice. Theoretical background and test cases on combustion of hydrocarbons are presented. PMID- 12546536 TI - The reliability of total citation rankings. AB - In short, citation analysis has become a small cottage industry today. The legitimacy of its use for a variety of purposes has been established. In fact, that legitimacy has extended to the point where many suspect uses of citations are accepted without significant skepticism. Rarely, if ever, are these counts, which compare one individual with another, accompanied by a set of caveats, about the limits of such individual comparisons or the bases on which the comparisons are being made (Jonathan R. Cole). (Cole, J. R. A Short History of the Use of Citations as a Measure of the Impact of Scientific and Scholarly Work, Chapter 14. In The Web of Knowledge; Cronin, B., Atkins, H. B., Eds.; ASIS Monograph Series, 2000; p 281.) PMID- 12546537 TI - Luddite: an information-theoretic library design tool. AB - We present an algorithm for the design of either combinatorial or discrete informative libraries. This approach is based on information theoretic techniques used extensively in coding theory. We have extended the information theoretic formalism to include an arbitrary number of property distribution constraints, such as Lipinski "drug-like" distributions. The method is demonstrated by comparing and contrasting a variety of different libraries selected from a single combinatorial source pool of compounds. PMID- 12546538 TI - Three-dimensional quantitative structure-property relationship (3D-QSPR) models for prediction of thermodynamic properties of polychlorinated biphenyls (PCBs): enthalpies of fusion and their application to estimates of enthalpies of sublimation and aqueous solubilities. AB - Comparative Molecular Field Analysis (CoMFA) has been used to develop three dimensional quantitative structure-property relationship (3D-QSPR) models for the fusion enthalpy at the melting point (Delta(fus)H(m)(T(fus))) of a representative set of polychlorinated biphenyls (PCBs). Various alignment schemes, such as inertial, as is, atom fit, and field fit, were used in this study to evaluate the predictive capabilities of the models. The CoMFA models have also been derived using partial atomic charges calculated from the electrostatic potential (ESP) and Gasteiger-Marsili (GM) methods. The combination of atom fit alignment and GM charges yielded the greatest self-consistency (r(2) = 0.955) and internal predictive ability (r(cv)(2) = 0.783). This CoMFA model was used to predict Delta(fus)H(m)(T(fus)) of the entire set of 209 PCB congeners, including 193 PCB congeners for which experimental values are unavailable. The CoMFA-predicted values, combined with previous estimations of vaporization and sublimation enthalpies, were used to construct a thermodynamic cycle that validated the internal self-consistency of the predictions for these three thermodynamic properties. The CoMFA-predicted values of fusion enthalpy were also used to calculate aqueous solubilities of PCBs using Mobile Order and Disorder Theory. The agreement between calculated and experimental values of solubility at 298.15 K, characterized by a standard deviation of +/- 0.41 log units, demonstrates the utility of CoMFA-predicted values of fusion enthalpies to calculate aqueous solubilities of PCBs. PMID- 12546539 TI - A measure of folding complexity for d-dimensional polymers. AB - A measure of folding characterizes aspects of the instantaneous organization of a polymer chain in space. For three-dimensional polymers (D = 3), one such measure is the mean overcrossing number. An intuitively similar property, the radial intersection number, has been proposed as a tool to characterize "folding features" in two-dimensional polymers (D = 2). In this work, we show rigorously that these measures are indeed related and that they can be derived as particular cases within a single, unified formulation. The present approach provides an analytical expression for a measure of folding complexity that can be applied to generic D-dimensional polymers. In the case D = 2, we show results for models derived from experimental structures by using optimized multidimensional scaling transformations for data compression. PMID- 12546540 TI - Many-body optimization using an ab initio monte carlo method. AB - Advances in computing power have made it possible to study solvated molecules using ab initio quantum chemistry. Inclusion of discrete solvent molecules is required to determine geometric information about solute/solvent clusters. Monte Carlo methods are well suited to finding minima in many-body systems, and ab initio methods are applicable to the widest range of systems. A first principles Monte Carlo (FPMC) method was developed to find minima in many-body systems, and emphasis was placed on implementing moves that increase the likelihood of finding minimum energy structures. Partial optimization and molecular interchange moves aid in finding minima and overcome the incomplete sampling that is unavoidable when using ab initio methods. FPMC was validated by studying the boron trifluoride-water system, and then the method was used to examine the methyl carbenium ion in water to demonstrate its application to solvation problems. PMID- 12546541 TI - What are the limits of applicability for graph theoretic descriptors in QSPR/QSAR? Modeling dipole moments of aromatic compounds with TOPS-MODE descriptors. AB - The numerous possibilities of using graph theoretic descriptors in QSPR/QSAR are analyzed, and some misunderstandings on the role of this theoretical approach in chemistry are clarified. Principal component analysis is used to obtain a property space for several physicochemical properties of aromatic compounds. It is proved that most of the QSPR applications of the graph-theoretic structure descriptors are concentrated to the description of properties in a very limited region of this property space. Here, we show that graph-theoretic approaches are also applicable to the modeling of physicochemical properties that are far away from this region traditionally studied. The molecular dipole moments of benzene derivatives, mono-, ortho-, meta-, and para-susbtituted, are modeled by using the Topological Sub-Structural Molecular Design (TOPS-MODE) approach. The TOPS-MODE approach used permits to calculate group dipole moments that are given for several substituents. The differences between these group dipoles and those obtained by simple difference between experimental values are analyzed. Some difficulties arising from this traditional way of deriving substituent constants are identified and analyzed. PMID- 12546542 TI - A fuzzy ARTMAP-based quantitative structure-property relationship (QSPR) for the Henry's law constant of organic compounds. AB - Quantitative structure-property relationships (QSPRs) for estimating a dimensionless Henry's Law constant of organic compounds at 25 degrees C were developed based on a fuzzy ARTMAP and back-propagation neural networks using a heterogeneous set of 495 organic compounds. A set of molecular descriptors developed from PM3 semiempirical MO-theory and topological descriptors (second order molecular connectivity index) were used as input parameters to the neural networks. Quantum chemical input descriptors included average molecular polarizability, dipole moments (total point charge, total hybridization, and total sum), ionization potential, and heat of formation. The fuzzy ARTMAP/QSPR correlated Henry's Law constant for -6.72 30 days) organ system morbidity for any secondary procedures. RESULTS: Mean length of follow-up for open patients was 20.1 +/- 17.1 months (range 1-150) compared to 12.4 +/- 9.6 months (range 1-60) after endovascular repair (p<0.05). There were 36 (12.1%) systemic complications after the primary open surgery and 15 (7.5%) after endovascular repair (p=NS). There were 43 (14.5%) combined primary and secondary morbidities in the open surgery group versus 15 (7.5%) for patients undergoing endovascular repair (p<0.01). The need for invasive procedures to treat these primary and secondary systemic complications was 4 times greater in the open group (17, 5.7%) than in endograft patients (3, 1.5%) (p<0.05). After secondary procedures (32 in the open group and 30 in the endovascular patients) for graft related complications, there were 7 (21.9%) adverse events in the open group versus none (0%) for endograft patients (p<0.01). Hospital lengths of stay following both primary and secondary procedures were lower for the endograft patients (p<0.01 and p<0.001, respectively). CONCLUSIONS: Endovascular stent graft repair compared to open surgery has reduced the early and late morbidity by half. Complications that require invasive or secondary surgical procedures and hospitalization are reduced with endovascular repair. PMID- 12546570 TI - The Zenith aortic stent-graft: a 5-year single-center experience. AB - PURPOSE: To evaluate the efficacy and midterm results of the Zenith stent-graft in the treatment of abdominal aortic aneurysms (AAA). METHODS: Since March 1994, 364 patients have undergone endovascular repair of infrarenal AAA. Of the 94 who were treated with the Zenith stent-graft from 1996 to 2002, 88 patients (82 men; mean age 72.6 +/- 6.5 years, range 47-88) with at least 6-month follow-up were analyzed. Sixty-one (69.3%) patients were considered at high risk for intervention; 7 ruptured AAAs were treated emergently. In all, 68 (77.3%) bifurcated stent-grafts (including 18 TriFab systems) and 20 aortomonoiliac configurations were used. Cumulative data on endoleak, migration, secondary procedures, and survival were evaluated with Kaplan-Meier analyses. RESULTS: Implantation success was 97.7%; 2 (2.3%) access-related failures were converted to open repair (1 immediate, 1 at 3 months). There were 3 (3.4%) graft limb thromboses (2 immediate, 1 late), 3 (3.4%) cases of colon ischemia due to embolization in 1 and hypogastric artery occlusion in 2, and 1 (1.1%) renal infarction due to embolism. Three (3.4%) patients died within 30 days. Eleven (12.5%) endoleaks and 1 (1.1%) late endograft migration were recorded. The 5-year cumulative endoleak and migration rates were 15% and 7%, respectively. Sixty three (71.6%) patients did not present any complication related to the repair during a mean follow-up of 20.6 +/- 14.9 months (range 6-68); notably, no complications were associated with the 18 TriFab systems. Six (6.8%) secondary procedures were performed (31% 5-year cumulative secondary procedural rate). All 6 (6.8%) aneurysm-related deaths (the 3 perioperative, 2 from late AAA rupture, and 1 during a secondary procedure) and 14 of 18 (20.4%) non-aneurysm-related deaths occurred in high-risk patients; the 5-year cumulative survival rates were 57% for any death and 92% for aneurysm-related deaths. CONCLUSIONS: The Zenith stent-graft appears both safe and effective in terms of midterm outcome of endovascular aortic aneurysm repair. PMID- 12546571 TI - Endovascular repair of acute AAAs under local anesthesia with bifurcated endografts: a feasibility study. AB - PURPOSE: To evaluate endovascular repair of abdominal aortic aneurysms (AAA) under local anesthesia in the acute setting. METHODS: Between 1998 and 2001, 47 patients with an acute AAA were evaluated for endovascular repair after informed consent, provided they were in a stable, albeit hypotensive condition. The patients underwent urgent computed tomography to assess suitability for endovascular repair; 16 were eligible for stent-graft repair: 9 were frank ruptures and 7 were symptomatic aneurysms. Complications and outcome of endovascular repair were evaluated; mortality was compared to a contemporaneous surgical cohort. RESULTS: Seven (23%) of 31 patients having a standard surgical procedure died in the study period compared to 1 (6%) of 16 patients undergoing endovascular repair (following conversion to surgery because of calcified access vessels). Twelve (75%) of the endovascular repairs were performed under local anesthesia; no complications with this mode of anesthesia were encountered. The median duration of the endovascular procedures was 110 minutes (range 75-240); median blood loss was 250 mL (range 100-2800 mL). Only 4 patients required blood transfusion, and only 8 patients required admission to the intensive care unit. There were 3 postoperative complications (1 ischemic colitis, 1 renal failure, 1 groin hematoma). During follow-up, 3 endograft patients received stent-graft extensions in uneventful procedures. Two patients died at 9 and 16 months from cardiac causes. CONCLUSIONS: This study demonstrates the feasibility and possible advantages of endovascular repair under local anesthesia in selected acute AAA patients. Further studies are needed to prove the advantages over open repair. PMID- 12546572 TI - Outcome of abdominal aortic endografting in high-risk patients: a 4-year single center study. AB - PURPOSE: To evaluate feasibility, safety, and effectiveness of endovascular abdominal aortic aneurysm (AAA) repair in patients whose fitness for surgery is questionable. METHODS: Between April 1997 and December 2001, 389 consecutive patients underwent endovascular AAA repair. Of these, 51 (13.1%) were ASA grade IV. The perioperative and late outcomes of this group were compared to the remaining 338 patients with ASA grades 0.05). Actuarial survival at 30 months was 62.9% in ASA IV group and 88.0% in ASA 0.05). The median time to graft migration was 14 months after the original operation (range 6-36). CONCLUSIONS: Distal device migration occurred frequently with the Nottingham system. Late graft migration was not associated with neck enlargement. Endografts with a suprarenal stent may have a decreased incidence of graft migration. PMID- 12546574 TI - Factors and forces influencing stent-graft migration after endovascular aortic aneurysm repair. AB - PURPOSE: To assess and validate the clinical features predisposing to stent-graft migration and to calculate the distal displacement forces exerted at the proximal fixation site following endovascular aortic aneurysm repair (EVAR). METHODS: Demographic, anatomical, and graft-related features from 2862 patients were analyzed in a regression model to identify variables associated with stent-graft migration, which was defined as device movement >5 mm or considered significant by the investigator. Using the principles of continuity and momentum, a mathematical model of blood flow was created. The pulse pressure, proximal aortic and distal iliac diameters, and the degree of iliac angulation were varied in the calculations, and the distal displacement force exerted at the proximal fixation site was calculated. RESULTS: Ninety-nine patients developed stent-graft migration, which was clinically relevant in 85 (3.0%). Hypertension (p=0.015), smoking (p=0.009), maximal aortic diameter (p=0.004), and distal transverse aortic diameter (p=0.03) correlated with migration in the univariate analysis, but iliac angulation did not quite achieve significance (p=0.06). On multivariate analysis, current smoking, hypertension, distal transverse aortic diameter, maximum common iliac diameter, and increasing proximal graft size were significantly associated with stent-graft migration. The mathematical model calculated the distal displacement force exerted on the proximal fixation site of the stent-graft and validated the clinical findings. The ratio of graft-diameter change from proximal aorta to distal iliac influenced the greatest increase in the displacement force. CONCLUSIONS: The mathematical model validated hypertension, aneurysm morphology, and endograft size as clinical factors significantly associated with stent-graft migration. These findings may have important implications for the choice and design of future stent-grafts. PMID- 12546575 TI - Successful treatment of aortic endograft thrombosis with rheolytic thrombectomy. AB - PURPOSE: To report the benefits of rheolytic thrombectomy for treating aortic endograft thrombosis. METHODS: Of 40 patients who received the Ancure bifurcated endograft to treat abdominal aortic aneurysm (AAA) during a 9-month period, 6 (15%) patients (6 men; mean age 62.6 years, range 53-77) developed thrombosis of the endograft at an average of 9 weeks (range 1-20 months). Five patients were taking aspirin, and 3 were on warfarin therapy for atrial fibrillation. Immediately after angiography, rheolytic thrombectomy was used to remove the thrombus, followed by adjunctive procedures to treat the underlying pathology. RESULTS: Causes were kinking or extrinsic compression of the graft limb in 5 cases and thrombosis of the surgical closure site in a common femoral artery. Mechanical thrombectomy was successful in restoring circulation in all cases; thrombolysis was used in 1. All 6 patients had additional stents placed in the graft limbs, re-establishing patency. There was no mortality or recurrent thrombosis in a follow-up that has extended to 26 months, but 1 patient required additional stenting for subsequent focal kinking of a graft limb above the previously implanted stent. CONCLUSIONS: Rheolytic thrombectomy can safely and effectively treat endograft thrombosis after endovascular AAA repair. Additional thrombolytic agents, angioplasty, and stenting may be needed to correct the underlying causes of the thrombosis. Prophylactic stenting of iliac limbs at the time of implantation in patients with complex anatomy may prevent thrombosis of unsupported bifurcated endografts. PMID- 12546576 TI - Aortomonoiliac stent-grafts for abdominal aortic aneurysm repair: association with iliofemoral crossover grafts. AB - PURPOSE: To analyze the outcome of endovascular exclusion of abdominal aortic aneurysm (AAA) using aortomonoiliac stent-grafts. METHODS: Fifty-seven consecutive patients (49 men; median age 70, range 56-89) with AAA >5 cm were treated in a 6-year period with the conical ELLA stent-graft. Forty-two (73.9%) patients were classified ASA (American Society of Anesthesiologists) IV and 6 as ASA V. In the majority of cases, the implantation procedure featured device delivery through the external iliac artery, transrenal placement of a bare stent in selected cases, and an iliofemoral crossover graft through a prevesical tunnel. RESULTS: Successful deployment was achieved in 56 (98.2%) patients. Mean time to discharge was 8.7 days (range 2-125). Two patients died in the 30-day period. Nine endoleaks occurred in 8 (14%) patients; 4 required further intervention. Mean follow-up was 35.3 months (range 1-66), during which 5 patients died from unrelated causes. No late endoleak, graft occlusion, device twisting/migration, or aneurysm rupture was observed. No correlation between type I endoleaks and unfavorable proximal neck or iliac artery anatomical characteristics could be found. Primary technical and clinical success rates were 86.0% and 94.7%, respectively. CONCLUSIONS: In this approach, the crossover graft remains in a retropubic space and consequently does not have all the disadvantages of a subcutaneously placed prosthesis. The results achieved in this group of high-risk patients support recommendation of this technique as a simple and safe alternative to bifurcated systems. PMID- 12546577 TI - Buttress technique to insure accurate placement of AneuRx stent-grafts in patients with complex anatomy. AB - PURPOSE: To describe a technique that prevents caudal displacement of the AneuRx stent-graft during removal of the nose cone and runners. TECHNIQUE: The buttress technique uses a 16-F sheath positioned from the contralateral femoral approach at the inferior edge of the endograft's main body to provide safe retraction of the runners and the nose cone and accurate endograft deployment. CONCLUSIONS: If accurate deployment of the stent-graft according to the manufacturer's directions is not possible, this buttress technique can insure proper positioning. It adds no additional time or equipment to the procedure, and it appears to be a safe and easily applied maneuver. PMID- 12546578 TI - Neurological outcomes after carotid stenting protected with the NeuroShield filter compared to unprotected stenting. AB - PURPOSE: To compare outcomes for two nonrandomized cohorts of patients with high grade carotid disease who underwent either unprotected carotid stenting or stent implantation protected by the NeuroShield filter. METHODS: Under this protocol, symptomatic patients with carotid stenoses >70% or asymptomatic patients with bilateral carotid stenoses who were being evaluated for coronary artery bypass grafting were eligible for carotid stenting. Between December 1998 and November 2001, 75 consecutive patients (57 men; median age 67 years range 45-85) underwent carotid stenting without cerebral protection; concurrently, 75 carotid stent procedures protected with the NeuroShield filter were performed in 73 patients (51 men; median age 66 years, range 47-83). A neurologist reviewed all patients before and after treatment. The groups were comparable for age, sex, and symptoms, but the protected group had a higher proportion of postsurgical restenoses (14.7% versus 1.3%; p=0.003). Outcome measures included death and neurological events at 24 hours and 30 days. RESULTS: There were minor technical difficulties in 12 of the protected group, but none were clinically relevant. The procedural all-stroke/death rates in the unprotected versus protected groups, respectively, were 5.3% (4/75) and 2.7% (2/75; p=0.681), while the disabling stroke/death rates were 4% (3/75) and 1.3% (1/75; p=0.620). At 30 days, the all stroke/death rates were 10.7% (8/75) in the unprotected group and 4.0% (3/75) in the protected group (p=0.117); the death/major-disability-from-stroke rates were 6.7% (5/75) and 2.7% (2/75), respectively (p=0.442). CONCLUSIONS: Filter-related complications are well tolerated. Neuroprotection devices have the potential to reduce the procedural neurological event rate. Larger series and/or randomized trials are required for further evaluation. PMID- 12546579 TI - Impact of cerebral protection devices on early outcome of carotid stenting. AB - PURPOSE: To evaluate the impact of cerebral protection devices on the procedural safety and outcome of carotid stent procedures. METHODS: From June 1997 to July 2001, 275 consecutive patients (208 men; mean age 71 +/- 7.4 years) underwent percutaneous angioplasty and/or stenting of the extracranial carotid artery. In the first 125 (45.4%) patients, the procedures were performed without cerebral protection. After January 2000, protection devices were routinely used (150 [54.6%] patients), including the Angioguard filter, GuardWire occlusion system, TRAP Vascular Filtration System, EPI Filter Wire, NeuroShield, Parodi Anti Embolism System, and Medicorp occlusive balloon. RESULTS: The percutaneous procedures were effective in 273 (99.3%) patients. No death or major stroke occurred in either group. In the unprotected group, 5 (4.0%) complications occurred: 3 (2.4%) minor strokes, 1 (0.8%) transient ischemic attack (TIA), and 1 (0.8%) subarachnoid hemorrhage. In the patients treated under cerebral protection, there were 2 (1.3%) complications: 1 (0.7%) minor stroke and 1 (0.7%) subarachnoid hemorrhage. There were 4 (3.2%) periprocedural embolic complications in the unprotected group versus 1 (0.7%) in the protected patients. CONCLUSIONS: Our data suggest that percutaneous dilation and stenting of the carotid arteries protected by cerebral protection devices is feasible and effective. In a consecutive series, the use of the cerebral protection systems reduced the acute neurological event rate related to embolic complications by 79%. PMID- 12546580 TI - In vitro comparison of four cerebral protection filters for preventing human plaque embolization during carotid interventions. AB - PURPOSE: To evaluate in an in vitro bench-top model the efficacy of 4 filtration devices designed for cerebral protection during carotid angioplasty. METHODS: Embolization during carotid angioplasty was simulated with human plaque material (8 to 12 particles weighing 6.02 +/- 0.10 mg) in an open flow model with the aortic arch and carotid bifurcation made from elastic silicone tubes and saline used as the fluid medium. The 500 to 1500-microm particles were injected into the internal carotid artery (ICA) in front of the test protection device, which was deployed 5 cm distal to the bifurcation. Particles getting past the protection device or flowing into the external carotid artery (ECA) were caught in 100 microm filters and weighed. Ten trials were made on each of 4 devices: Angioguard, FilterWire EX, TRAP, and NeuroShield. All were deployed according to the manufacturers' directions except the FilterWire, which was manually repositioned in all tests to eliminate a gap between the filter and the tube wall. RESULTS: The lowest weight of missed particles in the effluent was obtained with the NeuroShield (0.05 +/- 0.04 mg, 0.8% of injected particle weight) and FilterWire (0.08 +/- 0.05 mg, 1.3%; p=0.254 compared to NeuroShield, p=0.006 versus TRAP). The weight of particles missed by the TRAP device (0.16 +/- 0.06 mg, 2.6%; p<0.001 versus NeuroShield and Angioguard, p<0.05 compared to FilterWire) was higher, and the largest amount of missed particles was observed with the Angioguard filter (0.27 +/- 0.06 mg, 4.4%; p<0.001 compared to all). NeuroShield and FilterWire were significantly different (p<0.001) compared to Angioguard and TRAP in a pairwise analysis. No embolization into the ECA occurred. CONCLUSIONS: In vitro, none of the tested devices had the ability to completely prevent embolization into the ICA. Comparing current designs, the NeuroShield filter and the FilterWire EX captured the highest percentage of human particles in this in vitro model, probably due to their larger filter volume. PMID- 12546581 TI - Unprotected carotid artery stenting compared to carotid endarterectomy in a community setting. AB - PURPOSE: To evaluate the feasibility and safety of carotid artery stenting (CAS) performed by a dedicated multidisciplinary team in a community hospital. METHODS: Forty-nine patients (30 men; mean age 74.3 +/- 7.4 years, range 51-88) with 14 symptomatic and 35 asymptomatic >50% carotid stenoses were enrolled into the study and treated with self-expanding stents without cerebral protection. The patients were evaluated at 30 days for death, neurological sequelae, and length of stay. A contemporaneous group of 140 patients (80 men; mean age 72.3 +/- 7.7 years, range 43-89) undergoing carotid endarterectomy (CEA) was used for comparison of the outcome measures. Owing to crossover of failed CAS patients to the CEA group, the data were analyzed according to intention-to-treat and treatment-received. RESULTS: The only difference in baseline characteristics between the treatment and control groups was the greater frequency of bilateral disease in the CEA group (p=0.009). The CAS procedures were successful in 47 (96.0%) patients; 2 access failures were converted to endarterectomy, and 1 access-site pseudoaneurysm was treated surgically. There were no deaths or myocardial infarctions in either group. In the CAS group, there were no major and only 2 (4.1%) minor strokes compared to 4 (2.9%) major and 4 (2.9%) minor strokes in the CEA group (p>0.05). Median LOS was significantly lower in the CAS group (p<0.0001). CONCLUSIONS: Careful case selection and multidisciplinary collaboration facilitate the safe performance of carotid stenting in a community setting with acceptable early results. PMID- 12546582 TI - Correlation of periprocedural systolic blood pressure changes with neurological events in high-risk carotid stent patients. AB - PURPOSE: To examine retrospectively the magnitude of change in systolic blood pressure (SBP) during carotid artery stenting and its relationship to neurological events. METHODS: In a 5-year period ending October 2000, 60 patients (36 men; mean age 67 +/- 9 years) were enrolled in a study to evaluate stenting in symptomatic or asymptomatic high-risk patients with > or =70% carotid artery stenosis. The majority (48, 80%) of the patients were symptomatic. Five patients with bilateral lesions were treated in staged procedures. Sixteen (25%) of the 65 lesions were postsurgical stenoses and 12 (18%) were secondary to neck radiation therapy. RESULTS: There were 2 (3%) minor and 2 (3%) major strokes (94% procedural success), of which 1 was fatal. Six (9%) transient neurological events were recorded during balloon inflation. The mean SBP change during or after stenting in 55 cases without neurological events was 34 +/- 14 mmHg, while the patients with transient or permanent neurological events had significantly greater changes in SBP (107 +/- 31 mmHg [p<0.003] and 134 +/- 14 mmHg [p<0.001], respectively). Patients exhibiting neurological sequelae had significantly higher SBP before the procedure than those without complications (203 +/- 30 versus 165 +/- 23 mmHg, p<0.001). There were no neurological events in patients with a <50 mmHg change in SBP. CONCLUSIONS: Patients with severely elevated baseline SBP (>180 mmHg) may be at higher risk for hemodynamic instability and neurological events during carotid stenting. The greater the change in SBP, the more severe the neurological event seems to be, but further studies in a greater number of patients are needed to evaluate the potential causes of SBP fluctuations in an effort to avoid neurological events. PMID- 12546583 TI - Treatment of thoracic aortic dissection with stent-grafts: midterm results. AB - PURPOSE: To evaluate the early and midterm outcomes after stent-graft implantation for dissection in the descending thoracic aorta. METHODS: Sixteen custom-made endovascular stent-grafts were implanted in 15 patients (10 men; mean age 55.9 +/- 13.7 years, range 32-82) with descending thoracic aortic dissection. Indications for stent-graft implantation were persistent symptoms unresponsive to medical treatment or progressive enlargement of the false lumen. Clinical and imaging surveillance with computed tomography was performed within 1 month of the procedure and at 3 to 6-month intervals in follow-up. RESULTS: Endovascular stent graft implantation at the target site was successful in 14 (93%) patients; 1 device migrated, leaving the false lumen open to flow in the failed case. One (7%) patient who was treated emergently for rupture died suddenly 2 days after the procedure. Over an average follow-up of 31.5 +/- 23.8 months, 1 (7%) patient died and 2 (14%) patients underwent surgical treatment due to recurrent dissection. The remaining 10 patients showed complete thrombosis of the false lumen; in 3, the false lumen completely resolved. CONCLUSIONS: Endovascular stent graft implantation in descending thoracic aortic dissection is a feasible, safe, and effective treatment modality. However, further studies are necessary in a greater number of patients to determine if wider application of this minimally invasive procedure is justified. PMID- 12546584 TI - Complications after endovascular repair of type B aortic dissection. AB - PURPOSE: To outline the complications encountered after endoluminal treatment in patients with type B aortic dissection. METHODS: Between 1999 and 2001, 14 patients (12 men; mean age 60.3 years, range 39-79) with isolated type B aortic dissection (13 chronic, 1 acute) underwent aortic stent-grafting. Three patients with chronic dissection presented an acute clinical picture and were managed emergently. The left subclavian artery was intentionally covered by the prosthesis in 9 patients. Follow-up studies were performed at 6-month intervals. RESULTS: Stent-graft implantation was technically successful in all patients, but incomplete sealing (endoleak) of the entry site required additional proximal stent-graft implantation in 4. The left subclavian artery remained patent in 5 patients. Secondary conversion was required in 3 patients: 2 for acute type A dissection resulting from injury to the aortic arch by Talent endografts and a sustained hemorrhage (left hemothorax). In another patient, a secondary intramural hematoma subsided spontaneously. Anterior spinal artery syndrome in 1 patient persisted at 1 month. No bypass was necessary for the 9 patients with the covered left subclavian arteries. Mean follow-up was 14 months (range 1-23). CONCLUSIONS: Stent-grafting is feasible in patients with type B aortic dissection, although it is associated with a considerable rate of complications. Frank reporting of these sequelae for a variety of stent-grafts is of paramount importance to clarifying the limitations of the method. PMID- 12546585 TI - Endograft therapy for diseases of the descending thoracic aorta: results in 43 high-risk patients. AB - PURPOSE: To report an initial experience with endovascular stent-graft implantation for diseases of the descending thoracic aorta in high-risk patients. METHODS: Forty-three patients (28 men; mean age 67 years, range 17-82) with 16 descending thoracic aortic dissections, 14 aneurysms, 7 contained ruptures, 3 mycotic aneurysms, 2 posttraumatic pseudoaneurysms, and an aneurysm of an anomalous right subclavian artery were treated between June 1999 and July 2001. Twenty-three (53%) patients were treated emergently. RESULTS: There were no conversions to open repair, but 3 (7%) patients died during the first 30 days (pneumonia, multiorgan failure, and acute bowel ischemia). Thirteen (30%) patients suffered 18 major complications (8 strokes, paraplegia in 3, respiratory insufficiency in 6, and 1 renal failure). Of 7 (16%) endoleaks detected in the early postoperative period, 3 required additional stents, while the other 4 were treated conservatively. Follow-up, which averaged 19 +/- 6 months (median: 13; range 0-34), was 100% complete. Five (12%) patients died: 3 of aortic rupture at 34, 47, and 139 days, respectively, and 2 from heart failure at 3 and 15 months, respectively. No late migration or endoleaks have been detected in the remaining 35 patients; however, 1 (2%) patient showed progressive aortic dissection proximal to the stent-graft. In all other cases, the size of the aneurysm or the false lumen was unchanged or diminished. CONCLUSIONS: Treatment of descending thoracic aortic diseases with an endovascular approach has acceptable early mortality and morbidity in high-risk patients. In selected cases, stent-grafts may afford the best therapy. PMID- 12546586 TI - Side-branched modular endograft system for thoracoabdominal aortic aneurysm repair. AB - PURPOSE: To describe a side-branched modular endograft system that provides adequate visceral artery perfusion with perfect seal during thoracoabdominal aortic aneurysm (TAAA) repair. CASE REPORT: A 76-year-old man with a 57-mm TAAA involving the celiac artery was treated with a customized Talent endograft consisting of a 46-mm x 18-cm stented main body and a 6-mm x 30-mm nonstented Dacron side branch. The graft was delivered through a surgical exposure of the left common femoral artery. A 6-mm x 10-cm Hemobahn stent-graft was introduced in the 30-mm side branch from the aorta to the celiac trunk through a long 8-F sheath via the left brachial artery. The patient recovered uneventfully except for a mild reactive inflammatory syndrome. Postoperative computed tomography demonstrated total exclusion of the TAAA sac and good antegrade perfusion of the celiac and superior mesenteric arteries, which has been maintained at the 6-month follow-up. CONCLUSIONS: Endovascular treatment of TAAA is feasible with further technical refinements of available technology. PMID- 12546587 TI - Vascular endothelial growth factor (VEGF165) and its influence on angiogenesis versus arteriogenesis in different vascular beds. AB - PURPOSE: To use local gene delivery to determine any district-specific influence of vascular endothelial growth factor (VEGF(165)) on angiogenesis and arteriogenesis in arteries of distinct developmental origin. METHODS: Coronary and peripheral arteries were chronically occluded in 30 Pietrain pigs using a percutaneous approach and blinded stent-graft. DNA was delivered to the adventitia in dosages corresponding to 10% of the body weight-adapted amount used in clinical trials. The coronary arteries in 12 animals and the peripheral arteries in 12 animals were treated or used as controls (no occlusion or occlusion with transfection of the beta-galactosidase gene). Six additional animals were sacrificed at 1 or 3 weeks for expression analyses, while the other 24 animals were sacrificed at 5 months for expression analysis and histology. Angiography, polymerase chain reaction analyses, and immunohistochemistry were performed. RESULTS: Expression of the VEGF gene was observed at 1 and 3 weeks following application, while transfected DNA was detected up to 5 months. New collaterals formed around occluded coronary arteries (2.63 +/- 0.69 fold, p<0.05 versus 1.24 +/- 0.40 fold for peripheral arteries), and angiographic arterial area increase was more pronounced in coronary (2.49 +/- 0.59 fold, p<0.05) than peripheral arteries (1.49 +/- 0.05 fold). There was no collateralization surrounding occluded peripheral arteries, but new arterial branches were seen (2.0 +/- 0.28, p<0.05 versus 1.07 +/- 0.31 for coronary). CONCLUSIONS: The response to VEGF, whether it is predominantly angiogenesis or arteriogenesis, is dependent on the target vessel. These observed differences in the behavior of arteries may be related to their differing developmental origins, which may have important implications for future therapeutic strategies using VEGF in different vessels. PMID- 12546588 TI - Arterial stenting and overdilation: does it change wall mechanics in small caliber arteries? AB - PURPOSE: To evaluate changes in arterial wall mechanics induced by stent overdilation in the rabbit aorta. METHODS: Twenty New Zealand white rabbits had initial stent deployment (3-mm x 8-mm Multilink) at 10% overdilation. Group A (n=11) had no subsequent balloon expansion of the stent and Group B (n=9) had 30% overdilation of the stent. A noninvasive B-mode ultrasound examination coupled with image processing allowed the measurement of systolic and diastolic diameter and the calculation of diameter compliance (Cd) and distensibility coefficient (DC) as indexes of arterial wall biomechanics. Measurements were performed before stenting in the infrarenal aorta, after initial stenting, and after stent overdilation at 3 locations: upstream, at the stent level, and downstream from the stent. RESULTS: Cd was significantly lower in the stented aorta after initial stenting (p<0.0001) and after stent overdilation (p<0.0001) than before stenting. At the stent level, Cd and DC were significantly lower than downstream (p<0.0001) or upstream (p<0.0001) from the stent after initial stenting, as well as after stent overdilation. Downstream from the stent, Cd and DC were significantly lower after stent overdilation than before stenting (p<0.05). CONCLUSIONS: Endovascular stenting of the rabbit aorta produces a significant decrease in arterial wall compliance and distensibility. Stent overdilation is responsible for a slight additional decrease of compliance downstream from the stent. PMID- 12546589 TI - Renal atrophy in atherosclerotic renovascular disease: gradual changes 6 months after successful angioplasty. AB - OBJECTIVE: To assess renal morphology using spiral computed tomographic angiography (CTA) before and 6 months after angioplasty of unilateral atherosclerotic renal artery stenosis (RAS). METHODS: Spiral CTA scans from 14 hypertensive patients (7 men; mean age 57.4 +/- 13.1 years) with significant RAS were performed before and 6 months after angioplasty; renal length, mean cortical thickness (MCT), cortical area (CA), and medullary length (ML) were measured from the axial slices in the 14 contralateral and 14 poststenotic then revascularized kidneys. Blood pressure, creatinine clearance, and number of antihypertensive drugs were analyzed. RESULTS: At 6 months, the systolic blood pressure was significantly reduced (p=0.007), but the number of antihypertensive drugs and the creatinine clearance did not change significantly. Most morphological parameters before and after angioplasty were different in the contralateral versus poststenotic kidneys (renal length, p=0.01; MCT, p=0.01; ML, p=0.03; CA, p=0.008). After angioplasty, there was an 11% drop in cortical atrophy associated with a mean 4-mm increase in medullary length in the poststenotic/revascularized kidneys. The contralateral kidneys exhibited corticomedullary thinning after angioplasty. CONCLUSIONS: In atherosclerotic renal disease, cortical thinning could be a useful timesaving marker to assess the significance of the lesion and to evaluate associated distal lesions. The favorable blood pressure outcome supports the existence of reversible ischemic lesions, particularly in the medulla. Angioplasty appears to be useful for the poststenotic kidney, according to the morphological study. PMID- 12546590 TI - Duplex ultrasound-guided femoropopliteal angioplasty: initial and 12-month results from a case controlled study. AB - PURPOSE: To evaluate initial technical success, procedural complications, and 12 month patency of duplex-guided angioplasty compared to conventional fluoroscopically-guided procedures. METHODS: One hundred four patients (65 men; mean age 69 years) who underwent duplex-guided femoropopliteal angioplasty were compared to 104 patients undergoing fluoroscopically-guide procedures who were matched for age, sex, baseline ankle-brachial index (ABI), and length and grade of lesion. Patients were followed for 12 months, and restenosis was assessed by ABI and duplex sonography. RESULTS: Technical success was achieved in 88 (84.6%) patients from the duplex-guided group and in 102 (98.1%) control patients (p=0.001). Periprocedural complications occurred in 12.5% (n=13) and 18.3% (n=19), respectively (p=0.4). Contrast-induced transient renal impairment was observed in 7 (6.7%) patients in the fluoroscopic group. One hundred (96.1%) patients in the duplex and 102 (98.1%) patients in the fluoroscopic group completed the 12-month follow-up. Restenosis was found in 35 (39.8%) patients of the duplex group and in 38 (37.2%) patients of the fluoroscopic group (p=0.8). CONCLUSIONS: Technical success of duplex-guided procedures was significantly lower compared to fluoroscopic angioplasty; complications and 12-month patency were similar with both techniques. Duplex-guided angioplasty may be a feasible alternative, particularly for patients at high risk for contrast-induced complications. PMID- 12546591 TI - Percutaneous transluminal laser angioplasty versus balloon dilation for treatment of popliteal artery occlusions. AB - PURPOSE: To compare the immediate results, complication rates, and long-term outcomes of percutaneous transluminal laser angioplasty (PTLA) versus balloon dilation alone in the treatment of popliteal artery occlusions. METHODS: In a prospective nonrandomized study conducted between December 1994 and June 2000, 215 symptomatic patients with unilateral popliteal occlusions were treated with either dilation alone (88 patients: 52 men; mean age 62 years, range 48-83) or PTLA (127 patients: 70 men; mean age 64 years, range 49-86) using a 308-nm excimer laser followed by dilation. The average occlusion length was 10.4 cm (range 3-14). RESULTS: PTLA was successful in recanalizing 105 (82.7%) arteries, while the recanalization rate for dilation alone was only 70.4% (62/88; p=0.045). After a mean follow-up of 36 months (range 6-52), the primary and secondary patency rates were 21.7% and 50.8%, respectively, in patients with PTLA and 16.3% and 35.2% in the angioplasty group (p=0.762). The complication rates associated with both techniques were similar. CONCLUSIONS: Although initial recanalization may be better with PTLA, it does not appear to add any long-term benefit over balloon dilation alone. PMID- 12546592 TI - Vibrational angioplasty in the treatment of chronic infrapopliteal arterial occlusions: preliminary experience. AB - PURPOSE: To evaluate the safety and efficacy of vibrational angioplasty in chronic infrapopliteal arterial occlusions. METHODS: Twelve patients (9 men, aged 54 to 90 years) with 13 below-knee arterial chronic total occlusions were treated percutaneously using vibrational angioplasty. The occlusions were located in the anterior tibial artery (n=5), the tibioperoneal trunk (n=4), the peroneal artery (n=1), the posterior tibial artery (n=1), and in both the tibioperoneal trunk and peroneal artery (n=2). The length of the lesions ranged from 5 to 14 cm. RESULTS: Recanalization was successful in 12 (92.3%) lesions. In 1 case, the wire perforated the arterial wall; the procedure was abandoned without clinical sequelae. The time to cross the occlusions with the wire ranged from 6 to 19 minutes. No other complications were observed. Clinical follow-up ranged to 18 months. Ten patients with ulceration or gangrene demonstrated good wound healing, and pain was alleviated in all successfully treated patients. CONCLUSIONS: Vibrational angioplasty appears feasible as a means of safely recanalizing chronic total occlusions of the infrapopliteal arteries. Further experience should be acquired to assess its short- and long-term effects on this vascular territory. PMID- 12546593 TI - Endovascular treatment of a multimorbid patient with late AAA rupture after stent graft placement: 1-year follow-up. AB - PURPOSE: To report successful endovascular management of a ruptured abdominal aortic aneurysm (AAA) in a multimorbid patient 40 months after primary stent grafting. CASE REPORT: A 64-year-old man presented with hypotension, severe back pain, and abdominal distension. Immediate computed tomography revealed a proximal type I endoleak due to distal migration of the stent-graft with subsequent rupture of the aneurysm. The patient was hemodynamically unstable, and open surgery was refused because of severe comorbidities that were the indications for initial endovascular repair. The diameter of the proximal aneurysm neck required the use of a thoracic stent-graft that was overly long, which led to occlusion of the contralateral stent-graft limb supplying not only the left leg but also a left kidney transplant. A crossover bypass was implanted to revascularize both. CONCLUSION: Minimally invasive strategies, even when challenged by complex vascular reconstructions, offer the possibility of managing ruptured aortic aneurysms in patients unsuitable for open surgery. PMID- 12546594 TI - Aortic stent-graft infection due to a presumed aortoenteric fistula. AB - PURPOSE: To report a case of late stent-graft infection with aortoenteric fistula. CASE REPORT: A 76-year-old Colombian man received an AneuRx stent-graft for a 5.5-cm infrarenal AAA. The aneurysm sac progressively shrank until 22 months postoperatively, when an increase in diameter was noted on magnetic resonance imaging without evidence of endoleak or air in the sac. Two months prior, the patient had developed fever and an elevated white blood cell count; he underwent a 6-week course of intravenous antibiotics. Shortly thereafter, the fever recurred, along with progressive weight loss, which prompted admission. The computed tomographic scan showed no evidence of endoleak, but gas collection was seen anteriorly in the sac; aspirated material was positive for a variety of organisms. At surgery 23 months after stent-graft implantation, pronounced inflammatory reaction and scarring were seen around the graft in conjunction with evidence of a healed duodenal perforation, suggestive of an aortoenteric fistula. The excised stent-graft was intact; no deterioration was seen. The patient had a protracted recovery but has been afebrile and asymptomatic >1 year after stent graft explantation CONCLUSIONS: Close surveillance after endovascular AAA repair is essential to detect late leaks, secondary migration, endotension, structural failure, and infection with or without aortoenteric fistula. PMID- 12546595 TI - Internal iliac aneurysm rupture into the rectum following endovascular exclusion: an unusual cause of massive lower gastrointestinal bleeding. AB - PURPOSE: To report a rare iliorectal fistula following endovascular treatment of an internal iliac aneurysm. CASE REPORT: A 76-year-old man developed lower gastrointestinal bleeding 3 months after successful endovascular exclusion of a left internal iliac aneurysm with coil embolization, attempted stent-grafting, ligation of the distal external iliac artery, and a femorofemoral crossover bypass. Aortography showed no clear intestinal bleeding point, but demonstrated recanalization and continued perfusion of the aneurysm. At laparotomy, an iliorectal fistula was detected. The common iliac artery was ligated proximally, the aneurysm sac was opened, and the back-bleeding internal iliac artery branches were oversewn. The rectum was closed primarily. He made an uneventful recovery. CONCLUSIONS: An iliorectal fistula is an extremely rare and unlikely complication of coil occlusion of an iliac aneurysm. A high index of suspicion for the diagnosis is of paramount importance. Periodic imaging in these patients is required to detect recanalization and continuing aneurysm expansion. PMID- 12546596 TI - Perivascular inflammatory reaction to a Hemobahn stent-graft: diagnosis with 3D MR angiography. AB - PURPOSE: To present a rare perivascular inflammatory reaction to a commercially produced polytetrafluoroethylene-covered stent and demonstrate the utility of 3 dimensional (3D) magnetic resonance angiography (MRA) in the diagnosis of this phenomenon. CASE REPORT: Three weeks after percutaneous deployment of a Hemobahn stent-graft to treat a high-grade stenosis and aneurysm of the proximal left superficial femoral artery (SFA), a 70-year-old diabetic man developed fever, pain, and local swelling of the left thigh. Venous thrombosis was excluded by ultrasound imaging; a normal flow profile was seen in the left common and superficial femoral arteries. Blood analysis found elevated inflammatory markers. MRA revealed extensive soft-tissue edema and perivascular contrast enhancement around the left SFA, but the stent-graft was patent. The clinical symptoms resolved within 7 days after initiation of anti-inflammatory therapy. Follow-up MRA scans demonstrated significantly reduced inflammatory reaction over the next few months. CONCLUSIONS: Symptomatic perivascular inflammatory soft-tissue response to a stent-graft can be diagnosed with MR imaging studies. PMID- 12546597 TI - Acute thrombotic subclavian artery occlusion treated with a new rotational thrombectomy device. AB - PURPOSE: To report the use of a new rotational thrombectomy device for percutaneous thrombectomy of an acutely occluded subclavian artery. CASE REPORT: A 71-year-old woman with a history of multivessel coronary disease complained of sudden onset of pain at rest and paleness of the left arm. Duplex ultrasound showed a localized thrombotic occlusion of the postvertebral subclavian artery and another at the bifurcation of the brachial artery. After angiographic confirmation, the subclavian artery was recanalized with an 8-F Rotarex device via a percutaneous transfemoral access; the bifurcation of the brachial artery was recanalized by local thrombolysis (50 mg rtPA) because the thrombectomy device was too short to reach the occlusion. Follow-up examinations up to 1 year have shown normalized perfusion of the left arm. CONCLUSIONS: This new thrombectomy device is a useful tool for the percutaneous treatment of acute occlusion in the brachiocephalic arteries. PMID- 12546598 TI - Coil embolization of an aortic arch false aneurysm. AB - PURPOSE: To report the endovascular occlusion of an anastomotic false aneurysm of the ascending aorta. CASE REPORT: A 51-year-old patient developed an anastomotic false aneurysm at the level of the aortic arch after surgical replacement of the aortic root and arch for aortic dissection. The aneurysm expanded after 6 months and because of its saccular shape was treated by selective occlusion with detachable microcoils. Magnetic resonance angiography showed the stability of the occlusion after 2 years. CONCLUSIONS: Endovascular treatment with coils may be a good alternative to surgery for saccular anastomotic false aneurysms. PMID- 12546599 TI - Intravascular ultrasound-guided stenting for chronic abdominal aortic dissection. AB - PURPOSE: To describe how the combined use of duplex and intravascular ultrasound (IVUS) can assist in the evaluation and treatment of isolated abdominal aortic dissection without need for contrast angiography. CASE REPORT: A 78-year-old man presented with intermittent bilateral buttock and thigh claudication. Duplex ultrasound and contrast-enhanced computed tomography (CT) confirmed a chronic dissection along 3 to 4 cm of the infrarenal abdominal aorta. During Extra Large Palmaz stent implantation, the procedure was based on IVUS images and fluoroscopy without angiography. Both duplex and IVUS images were critical in assessing the type and extent of the lesion to be treated, in guiding the procedure, and in assessing its satisfactory outcome. CONCLUSIONS: In selected cases, ultrasound based imaging modalities can provide most of the information required to accomplish complex aortic procedures. PMID- 12546600 TI - Percutaneous infusion system for in situ thrombolysis in the abdominal aorta and iliac arteries. AB - PURPOSE: To report successful treatment of an occluded aorta with a new percutaneous infusion system. CASE REPORT: A 57-year-old man was admitted in severe pain with cyanosis and acrocyanotic pigmentation in the lower abdomen and limbs. The patient had symptoms of renal disease and severe oligoanuria. Ultrasound scanning detected no flow in the lower limbs, and angiography identified total thromboembolic occlusion of the aorta immediately below the origin of the renal arteries. A new Trellis percutaneous thrombolytic infusion system was used for controlled site-specific infusion and dispersion of streptokinase into the abdominal aorta and the iliofemoral arteries bilaterally with successful restoration of flow and no complications. CONCLUSIONS: The Trellis system is effective for rapid lysis of thrombotic emboli using low doses of lytic agents. The unique design of the Trellis allowed complete aspiration of the residual thrombus, thus avoiding adverse effects. PMID- 12546601 TI - Multiple stenting in Takayasu arteritis. AB - PURPOSE: To illustrate the possible peri- and postprocedural complications of stent treatment for Takayasu arteritis and suggest ways of reducing these risks. CASE REPORT: A 69-year-old woman with Takayasu arteritis had multiple stents implanted in the aortic arch (3 Memotherm) and main branches of the thoracic aorta (individual Corinthian stents in the subclavian and brachiocephalic arteries) following balloon dilation. She suffered reperfusion injury with cerebral hemorrhage following the procedure, preventing the use of anticoagulation or antiplatelet therapy. Further cerebral infarctions/hemorrhages over the ensuing months led to her death. CONCLUSIONS: Severe vascular disease of any cause confers a poor prognosis. Endovascular stenting may seem an attractive option for management of these patients, but there is often significant morbidity and mortality associated with the underlying disease as well as the stent. PMID- 12546602 TI - Re: endovascular versus conventional open repair of acute abdominal aortic aneurysm: feasibility and preliminary results. PMID- 12546604 TI - Medicine and professionalism. PMID- 12546605 TI - Etiologic considerations in the patient with syncope and an apparently normal heart. PMID- 12546606 TI - The prevalence of myocarditis and skeletal muscle injury during acute viral infection in adults: measurement of cardiac troponins I and T in 152 patients with acute influenza infection. AB - BACKGROUND: Current literature suggests that myocarditis is a common event during influenza infection, occurring with a prevalence rate of up to 10%, but these studies have relied on relatively nonspecific techniques of varying sensitivities for the detection of myocyte injury. Using measurement of cardiac troponins I and T, this study sought to determine the prevalence of myocarditis in a large unselected cohort of patients with serologically confirmed acute influenza infection. METHODS: A total of 152 subjects were recruited from 60 primary care and university health centers. Serial creatine kinase (CK), CK-MB, and cardiac troponin I and T measurements were taken on days 1, 6, and 21 following presentation. RESULTS: Creatine kinase levels were elevated (mean +/- SD levels, 830 +/- 1531 U/L; range, 181-7280 U/L) during the collection period in 18 patients (12%). Twelve (67%) of these had elevated CK levels on day 1 of presentation. Overall CK-MB levels were higher than 25 U/L in 3 patients with elevated CK readings but in no patient was the CK-MB fraction greater than 6%. Cardiac troponin I and T levels were not raised in any of the patients. CONCLUSIONS: Using more sensitive and specific markers of myocardial injury, we demonstrate that the prevalence of myocarditis during acute influenza infection is substantially lower than previously thought, whereas skeletal muscle injury is relatively common. Although we were unable to conclude that no myocardial inflammation was present, it seems likely that this complication is rare. PMID- 12546607 TI - Lack of efficacy of acetaminophen in treating symptomatic knee osteoarthritis: a randomized, double-blind, placebo-controlled comparison trial with diclofenac sodium. AB - BACKGROUND: Recommendations state that acetaminophen should be used in preference to nonsteroidal anti-inflammatory drugs in the initial treatment of symptomatic osteoarthritis (OA) of the hip or knee, because of lesser toxicity and the pervasive belief that acetaminophen is not only effective in treating OA pain but is of equal analgesic efficacy as nonsteroidal anti-inflammatory drugs. METHODS: This was a randomized, double-blind, placebo-controlled trial of diclofenac sodium, 75 mg twice daily, vs acetaminophen, 1000 mg 4 times daily, in 82 subjects with symptomatic OA of the medial knee. Osteoarthritis was quantitated radiographically, and subjects met stringent baseline pain criteria. The primary evaluation of efficacy used the Western Ontario and McMaster Universities Osteoarthritis Index, with evaluations at screening, baseline, and 2 and 12 weeks after treatment. Intention-to-treat analysis was used. RESULTS: Twenty-five subjects were randomized to diclofenac, 29 to acetaminophen, and 28 to placebo. The groups were closely matched for age, sex, body mass index, prior use of OA medications, baseline pain, and radiographic features. At 2 and 12 weeks, clinically and statistically significant (P<.001) improvements were seen in the diclofenac-treated group; however, no significant improvements were seen in the acetaminophen-treated group (P =.92 at 2 weeks and.19 at 12 weeks). Stratification of subjects according to baseline pain, prestudy OA medication, and radiographic grade showed no clear pattern of preferential response to diclofenac, and did not reveal a subset of subjects who responded to acetaminophen. CONCLUSIONS: Diclofenac is effective in the symptomatic treatment of OA of the knee, but acetaminophen is not. A review of the literature reveals that there is scanty published evidence for a therapeutic effect of acetaminophen relative to placebo in patients with OA of the knee, because most published studies use active comparators (ie, nonsteroidal anti-inflammatory drugs) only. The advocacy of acetaminophen use in subjects with OA of the knee should be reconsidered pending further placebo-controlled studies. PMID- 12546608 TI - Prevalence and risk factors for carriage of methicillin-resistant Staphylococcus aureus at admission to the intensive care unit: results of a multicenter study. AB - BACKGROUND: Detection of methicillin-resistant Staphylococcus aureus (MRSA) carriers on admission to the intensive care unit (ICU) is an important component of strategies for controlling the spread of MRSA. METHODS: A prospective multicenter study was conducted in 14 French ICUs for 6 months. All patients were screened within 24 hours after admission, using nasal and cutaneous swabs In addition, clinical samples were obtained. Patient data collected on ICU admission included presence of immunosuppression; history of hospital stay, surgery, antimicrobial treatments, or previous colonization with MRSA; chronic health evaluation and McCabe scores; reason for admission; whether the patient was transferred from another ward; severity of illness; presence of skin lesions; and invasive procedures. Risk factors for MRSA carriage at ICU admission were estimated, and significantly associated variables were used to develop a predictive score for MRSA carriage. A cost-benefit analysis was then performed. RESULTS: Of the 2347 admissions with MRSA screening, 162 (6.9%; range, 3.7%-20.0% among ICUs) were positive for MRSA, of whom 54.3% were detected through screening specimens only. Of the 2310 first admissions (vs repeat admissions) to the ICU, 96 were newly identified MRSA carriers. Factors associated with MRSA carriage in the multivariate analysis were age older than 60 years, prolonged hospital stay in transferred patients, history of hospitalization or surgery, and presence of open skin lesions in directly admitted patients. Only universal screening detected MRSA carriage with acceptable sensitivity. A cost-benefit analysis confirmed that universal screening and preventive isolation were beneficial. CONCLUSIONS: The prevalence of MRSA carriage on admission to the ICU is high in this endemic setting. Screening for MRSA on admission is useful to identify the imported cases and should be performed in all ICU-admitted patients. PMID- 12546609 TI - Genomes, transcriptomes, and proteomes: molecular medicine and its impact on medical practice. AB - The human genome project and the technological breakthroughs it has produced have moved the field of molecular medicine forward with breathtaking speed. This will impact not only the advance of scientific discoveries and the way science is conducted but also the clinical practice of medicine. In this review we explain the basic principles of these new technologies. Their potential use and impact are demonstrated by using diabetes mellitus as an example of a common and serious medical disorder. Finally, several potentially adverse consequences of "excessive" knowledge are discussed. PMID- 12546610 TI - Effect of a computerized alert on the management of hypokalemia in hospitalized patients. AB - BACKGROUND: Electrolyte abnormalities, including hypokalemia, are frequently encountered among hospitalized patients. Their management, when carefully audited, reveals major shortcomings. OBJECTIVE: To evaluate the effect of a computerized alert on the management of severe hypokalemia in hospitalized patients. METHODS: All patients who experienced severe hypokalemia (serum potassium levels <3.0 mEq/L) during their hospitalization at Hadassah Medical Center (a 1000-bed teaching institution on 2 campuses in Jerusalem, Israel) were included in the study. The study intervention was a computerized alert consisting of a flashing screen or printed warning for patients with serum potassium levels below 3.0 mEq/L, visible whenever an individual patient's or entire ward's results were accessed on any hospital computer. Using a previously validated computerized audit technique, we analyzed the management of hypokalemia 6 months before and 6 months after implementation of the alert intervention. RESULTS: Comparing outcomes before and after the intervention, nonmeasurement of a subsequent serum potassium level after an initial low value decreased by 36.1% (P =.08). Failure to correct the serum potassium level to above 3.5 mEq/L during the hospitalization decreased by 28.6% (P =.02). Discharge from the hospital with a subnormal serum potassium level decreased by 17.2% (P =.06). CONCLUSIONS: A computerized alert system improved the management of hypokalemia in a tertiary care hospital. This was achieved at minimal cost and with no evidence of harm. The computerized audit based on a laboratory information system is an efficient tool for evaluating this intervention. PMID- 12546611 TI - A prospective study of sleep duration and coronary heart disease in women. AB - BACKGROUND: Long-term sleep deprivation is common in today's society. Recent experiments have demonstrated that short-term sleep deprivation in healthy subjects results in adverse physiologic changes, including a decreased glucose tolerance and an increased blood pressure. However, the long-term health consequences of long-term sleep deprivation are unclear. The objective of this study was to determine whether decreased sleep duration (from self-reports) is associated with an increased risk of coronary events. METHODS: We studied a cohort of 71 617 US female health professionals (aged 45-65 years), without reported coronary heart disease (CHD) at baseline, who were enrolled in the Nurses' Health Study. Subjects were mailed a questionnaire in 1986 asking about daily sleep duration. Subjects were followed up until June 30, 1996, for the occurrence of CHD-related events. We assessed the relationship between self reported sleep duration and incident CHD. RESULTS: A total of 934 coronary events were documented (271 fatal and 663 nonfatal) during the 10 years of follow up. Age-adjusted relative risks (95% confidence intervals) of CHD (with 8 hours of daily sleep being considered the reference group) for individuals reporting 5 or fewer, 6, and 7 hours of sleep were 1.82 (1.34-2.41), 1.30 (1.08-1.57), and 1.06 (0.89-1.26), respectively. The relative risk (95% confidence interval) for 9 or more hours of sleep was 1.57 (1.18-2.11). After adjusting for various potential confounders, including snoring, body mass index, and smoking, the relative risks of CHD (95% confidence intervals) for individuals reporting 5 or fewer, 6, and 7 hours of sleep were 1.45 (1.10-1.92), 1.18 (0.98-1.42), and 1.09 (0.91-1.30), respectively. The relative risk (95% confidence interval) for 9 or more hours of sleep was 1.38 (1.03-1.86). CONCLUSION: Short and long self-reported sleep durations are independently associated with a modestly increased risk of coronary events. PMID- 12546612 TI - Hypertension in acute ischemic stroke: a compensatory mechanism or an additional damaging factor? AB - BACKGROUND: In acute ischemic stroke, a transient blood pressure (BP) elevation is common, but the best management is still unknown. Therefore, we investigated retrospectively the relationship between BP after ischemic stroke and neurological outcome (evaluated by means of the National Institutes of Health Stroke Scale score at day 7). METHODS: The medical records of 92 consecutive patients with acute ischemic stroke, aged 47 to 96 years, were examined. Blood pressure was measured on admission, 4 times during the first 24 hours, 3 times daily for the first 4 days, and twice daily on day 7 (or at discharge). Antihypertensive treatment was given according to American Heart Association guidelines. RESULTS: The region damaged by the stroke was total anterior in 16 patients (17%), partial anterior in 30 (33%), lacunar in 34 (37%), and posterior circulation in 12 (13%). Stroke pathogenesis was cardioembolic in 28 (30%), atherothrombotic in 29 (32%), and lacunar in 34 (37%). The systolic BP range was 140 to 220 mm Hg; diastolic BP, 70 to 110 mm Hg. Initial BP was higher in the group with lacunar infarction than in the other groups (P<.05). The patients with the best outcome had the highest BP during the first 24 hours. The neurological outcome was strongly influenced by baseline stroke severity (NIH Scale score) and admission BP. Better initial neurological conditions and higher initial BP resulted in better neurological outcomes. CONCLUSIONS: The outcome of stroke is influenced by the type of stroke and initial BP. Lacunar stroke and the highest BP on admission carry the best prognosis, whereas the reverse is true for posterior circulation infarction and low BP. We found no evidence that, within the present BP range, hypertension is harmful and that its lowering is beneficial. PMID- 12546613 TI - Validity and clinical utility of the aspartate aminotransferase-alanine aminotransferase ratio in assessing disease severity and prognosis in patients with hepatitis C virus-related chronic liver disease. AB - BACKGROUND: The aspartate aminotransferase-alanine aminotransferase ratio (AST/ALT ratio) has been used to noninvasively assess the severity of disease in patients with chronic liver disease (CLD). We previously demonstrated that progressive liver functional impairment is associated with an increase in the AST/ALT ratio. OBJECTIVES: To evaluate the reproducibility and transportability of the AST/ALT ratio in a large cohort of patients with different degrees of hepatitis C virus (HCV)-related CLD, to confirm the correlation between progressive impairment of liver function and increase in the AST/ALT ratio, to evaluate whether diagnostic accuracy of the ALT/AST ratio can be improved by using it with other biochemical variables, and to assess the 1-year prognostic capability of the AST/ALT ratio in patients with liver cirrhosis. PATIENTS AND METHODS: We retrospectively evaluated 252 patients with HCV-related CLD. The AST/ALT ratio was correlated with the degree of liver fibrosis in patients with chronic hepatitis and with the Child-Pugh score in patients with cirrhosis. All patients had undergone monoethylglycinexylidide (MEGX) testing to evaluate liver function. We assessed the prognostic ability of the AST/ALT ratio in a subset of 63 cirrhotic patients who were followed up for at least 1 year. RESULTS: The AST/ALT ratio was more frequently 1 or higher in cirrhotic patients (P<.001). There was a significant correlation between MEGX values and the AST/ALT ratio (r(s) = -0.621, P<.001). Multivariate stepwise logistic analysis showed that AST/ALT ratio, platelet count (PLT), MEGX values, and prothrombin activity were independently associated with the presence of cirrhosis. Combined assessment of the AST/ALT ratio and/or PLT obtained 97.0% positive predictive value and 97.9% negative predictive value for the diagnosis of cirrhosis. The AST/ALT ratio had 81.3% sensitivity and 55.3% specificity in identifying cirrhotic patients who died within 1-year of follow-up. CONCLUSIONS: The AST/ALT ratio is both reproducible and transportable in patients with HCV-related CLD. The AST/ALT ratio is correlated with both histologic stage and clinical evaluation. Progressive liver functional impairment is reflected by an increase in the AST/ALT ratio. Noninvasive evaluation by means of the combined AST/ALT ratio and PLT assessment misclassifies only a few cirrhotic patients. In cirrhotic patients, the AST/ALT ratio provides medium-term prognostic information that is no different from that provided by established prognostic scores. PMID- 12546614 TI - Outcome and treatment of Bartonella endocarditis. AB - BACKGROUND: Endocarditis caused by Bartonella species is a potentially lethal infection characterized by a subacute evolution and severe valvular lesions. OBJECTIVES: To evaluate the outcome of patients with Bartonella endocarditis and to define the best antibiotic regimen using the following measures: recovery, relapse, or death. METHODS: We performed a retrospective study on 101 patients who were diagnosed in our laboratory as having Bartonella endocarditis between January 1, 1995, and April 30, 2001. Bartonella infection was diagnosed using immunofluorescence with a 1:800 cutoff, polymerase chain reaction amplification of DNA, and/or culture findings of Bartonella species from whole blood, serum, and/or valvular biopsy specimens. A standardized questionnaire was completed by investigators for each patient. RESULTS: Twelve of the 101 patients died and 2 relapsed. Patients receiving an aminoglycoside were more likely to fully recover (P =.02), and those treated with aminoglycosides for at least 14 days were more likely to survive than those with shorter therapy duration (P =.02). CONCLUSION: Effective antibiotic therapy for Bartonella endocarditis should include an aminoglycoside prescribed for a minimum of 2 weeks. PMID- 12546615 TI - Lower physician estimate of underlying asthma severity leads to undertreatment. AB - BACKGROUND: Asthma undertreatment has been linked to poor outcomes. National guidelines recommend that physicians classify asthma severity based on pretreatment symptoms and titrate care as the disease changes in the individual patient. This study evaluated the extent to which the physician estimate of underlying severity affects a patient's asthma care. METHODS: Data used were collected from a cohort of adults with asthma enrolled in managed care. Eligible patients were adults enrolled in managed care with medical encounters coded for asthma. Physicians were eligible if they were main asthma providers. The patient survey covered demographics, symptoms, asthma treatment, and self-management knowledge. Physicians were asked to assess the underlying severity of their patients' asthma. RESULTS: There were 4005 patients with asthma with physician estimates of underlying severity. Of the patients, 70.1% were female (mean age, 44.8 years) and 83.5% were white. Most patients' current asthma symptoms were moderate (39.4%) and severe (50.1%). Most physician estimates of underlying severity were mild (44.6%) and moderate (44.5%). Among those patients reporting moderate symptoms, daily inhaled corticosteroid use was reported in 35.2% when physician estimates were mild, 53.0% when moderate, and 68.1% when severe (P =.001). Rates of peak flowmeter ownership, allergy testing, and self-management knowledge tracked similarly with physician estimates of underlying severity. CONCLUSIONS: Physician estimates of underlying asthma severity appear to determine asthma care. For patients with inadequate symptom control, lower physician estimates of underlying severity were associated with care that is less consistent with national guidelines. To improve the quality of asthma care, physicians need to update treatment based on their patients' current symptoms and adapt care accordingly. PMID- 12546616 TI - Passive transfer of nut allergy after liver transplantation. AB - An anaphylactic reaction to cashew nut developed in a nonatopic 60-year-old man 25 days after receiving a liver allograft from a 15-year-old atopic boy who died of anaphylaxis after peanut ingestion. The liver recipient had no history of nut allergy. Posttransplantation skin prick test results were positive for peanut, cashew nut, and sesame seed, and the donor had allergen-specific IgE antibodies to the same 3 allergens. Contact tracing of the recipients of other solid organs from the same donor disclosed no other development of allergic symptoms after ingestion of peanut or cashew nut. Results of molecular HLA typing did not detect any donor-origin leukocytes in the recipient after transplantation, which excluded peripheral microchimerism. The patient inadvertently ingested peanut contaminated food and suffered a second anaphylactic reaction 32 weeks after the transplantation. This case illustrates that transfer of IgE-mediated hypersensitivity can occur after liver transplantation and have potentially serious consequences. We therefore recommend that organ donors undergo screening for allergies, and that recipients be advised regarding allergen avoidance. PMID- 12546617 TI - Are patients who develop angioedema with ACE inhibition at risk of the same problem with AT1 receptor blockers? PMID- 12546618 TI - C-reactive protein screening for cardiovascular disease. PMID- 12546619 TI - Ephebiatrics is adolescent medicine. PMID- 12546620 TI - D-dimer tests for assessment of patients with suspected pulmonary embolism. PMID- 12546621 TI - Physician career satisfaction across specialties: are we getting the true picture? PMID- 12546623 TI - Alcohol and acetaminophen hepatotoxicity. PMID- 12546624 TI - The new definition of myocardial infarction. PMID- 12546626 TI - Plasma D-dimer test accuracy can be affected by heparin administration. PMID- 12546628 TI - Potency of inhaled corticosteroid fails to predict reduced emergency department visits. PMID- 12546630 TI - Cowpox--not cows but cats. PMID- 12546631 TI - Elevated beta2-adrenoceptor protein concentration in adipose tissue from obese subjects is closely related to the body mass index and waist/hip ratio. AB - The aim of the present study was to quantify beta(2)-adrenoceptor protein content in adipose tissue during fasting, and to study the relationships between beta(2) adrenoceptor protein and mRNA levels and changes in metabolites related to lipolysis. Groups of male subjects with a body mass index of <25 kg/m(2) or >30 kg/m(2) fasted for 60 h. Abdominal subcutaneous fat biopsies were analysed for receptor mRNA levels by reverse transcription-PCR-HPLC. The beta(2)-adrenoceptor protein concentration was measured by Western blotting using fluorescence laser scanning for detection. The beta(2)-adrenoceptor protein concentration per cell (on a DNA basis) was higher in obese subjects ( P <0.03). There were highly significant relationships between beta(2)-adrenoceptor protein concentration and both body mass index and waist/hip ratio ( P <0.001 for both). Furthermore, there was an inverse relationship between the receptor protein concentration and the serum beta-hydroxybutyrate level during fasting ( P <0.005). beta(2)-Adrenoceptor protein levels decreased in both groups during fasting, to a similar degree. Basal beta(2)-adrenoceptor mRNA levels were similar in the two groups, but there was a smaller increase in the obese group during fasting ( P <0.03). The increased beta(2)-adrenoceptor protein level in obese subjects is likely to be related to the greater plasma membrane area of their adipocytes. The decrease during fasting may be due to increased binding of noradrenaline and subsequent internalization and degradation of the receptor. Elevated levels of less responsive beta(2)-adrenoceptor protein in obese subjects may contribute to the development of obesity. PMID- 12546632 TI - Urinary microalbumin/creatinine ratios: reference range in uncomplicated pregnancy. AB - During uncomplicated pregnancy, the development of proteinuria is accepted as a poor prognostic sign and is associated with increasing maternal and perinatal mortality and morbidity. Physiological proteinuria increases with increasing gestation and one of its largest constituents is albumin. The reference range for the (micro)albumin/creatinine ratio (ACR) has not been described for normal pregnancy. This prospective cross-sectional study describes the gestation specific 95% reference ranges for urinary microalbumin concentration, creatinine concentration and ACR in uncomplicated pregnancy. There is a significant increase ( P =0.016) in the ACR in the third trimester. The mean difference is 0.091 mg of albumin/mmol of creatinine (95% confidence interval, 0.014-0.168). Our results describe the first well-defined gestation-specific 95% reference range for a point-of-care measurement of the ACR. These data are essential if such testing is to be employed in antenatal care. PMID- 12546633 TI - Effects of angiotensin II receptor blockade versus angiotensin-converting-enzyme inhibition on ventricular remodelling following myocardial infarction in the mouse. AB - Left ventricular (LV) remodelling following myocardial infarction (MI) is associated with increased morbidity and mortality. Previous data suggest that angiotensin II (Ang II) plays a central role in the molecular events contributing to LV remodelling. We explored the effects of angiotensin-converting-enzyme (ACE) inhibition versus Ang II (AT(1)) receptor blockade on LV remodelling in mice post MI. Mice underwent sham procedure or left coronary artery ligation, and received placebo, the AT(1) receptor antagonist, losartan or the ACE inhibitor, enalapril. At 6 weeks, echocardiography and haemodynamic studies were performed. Infarct size and interstitial collagen content were determined. Expression of genes encoding atrial natriuretic peptide (ANP), collagen type I, AT(1a) and AT(1b) receptors were measured. The placebo MI group showed increased LV end-diastolic diameter, LV end-systolic diameter with depressed fractional shortening ( P <0.01 versus shams), increased LV mass and volume (both P <0.01 versus shams). The placebo MI group also exhibited increased non-infarct zone collagen content ( P <0.01), ANP ( P <0.01) and collagen type 1 ( P <0.01) gene expression, with a non significant rise in AT(1a) receptor gene expression. Neither losartan or enalapril prevented LV dilation or improved fractional shortening. Both similarly lowered systolic blood pressure ( P <0.01 for each versus placebo). Losartan and enalapril inhibited LV hypertrophy ( P <0.01), and decreased ANP ( P <0.01) and collagen type 1 gene expression ( P <0.05). Levels of AT(1a) receptor gene expression were higher than shams ( P <0.05 for both), but similar to placebo. AT(1b) receptor gene expression was much lower than that for AT(1a) receptor and similar in all groups. Thus, in this model, AT(1) receptor antagonism and ACE inhibition have equivalent inhibitory effects on myocardial hypertrophy and fibrosis. These results serve as an important basis for planned investigations to evaluate the anti-remodelling effects of these agents on mice in which genetic manipulations are used to disrupt components of the Ang II signalling system. PMID- 12546634 TI - Changes in the transthoracic impedance signal predict the outcome of a 70 degrees head-up tilt test. AB - We determined whether early changes in central haemodynamics, as determined by transthoracic impedance, induced by a 70 degrees head-up tilt (HUT) test could predict syncope. Heart rate, arterial blood pressure and central haemodynamics [pre-ejection period and rapid left ventricular ejection time ( T (1)), slow ejection time ( T (2)) and d Z /d t (max) (where Z is thoracic impedance), assessed by the transthoracic impedance technique], were recorded during supine rest and during a 45 min 70 degrees HUT test in 68 patients (40+/-2 years) with a history of unexplained recurrent syncope. We found that 38 patients (42+/-3 years) had a symptomatic outcome to 70 degrees HUT (fainters) and 30 (39+/-2 years) had a negative outcome (non-fainters). When measured between 5 and 10 min of 70 degrees HUT, T (2) had increased significantly only in the fainters, and a change in T (2) of >40 ms from baseline predicted a positive outcome with a sensitivity of 68% and a specificity of 70%. During supine rest prior to 70 degrees HUT, the fainters exhibited a shorter T (2) than non-fainters (183+/-10 compared with 233+/-14 ms; P <0.01), and a T (2) of <199 ms predicted a positive outcome to 70 degrees HUT with a sensitivity of 68% and a specificity of 63%. Incorporation of the changes that occurred from rest to 70 degrees HUT in other haemodynamic variables (heart rate >11 beats/min, systolic pressure <2 mmHg, diastolic pressure <7 mmHg and pulse pressure <-3 mmHg) increased the specificity to 97% and the positive predictive value to 93%. Thus transthoracic impedance could detect differences in central haemodynamics between fainters and non fainters during supine rest and during the initial period of 70 degrees HUT with a consistent sensitivity and specificity when combined with peripheral haemodynamic variables. PMID- 12546635 TI - Aspects of organ protein, amino acid and glucose metabolism in a porcine model of hypermetabolic sepsis. AB - Although glucose and protein metabolism have been investigated extensively in experimental models of hypodynamic sepsis, relatively little information is available regarding the compensated stage of sepsis. We investigated interorgan amino acid and glucose metabolism in a porcine model of compensated hyperdynamic sepsis. Fasting catheterized pigs received endotoxin ( Escherichia coli lipopolysaccharide; 3 microg.h(-1).kg(-1); intravenous) or saline (controls) and volume resuscitation over 24 h to reproduce hyperdynamic sepsis. Primed-constant infusions of p -aminohippurate and (3)H-labelled isotopes were used to measure glucose, amino acid and protein metabolism across the portal-drained viscera, liver and hindquarters (to represent muscle) at 0 and 24 h of endotoxaemia. Whole body protein and glucose flux were increased during hyperdynamic compensated sepsis. In endotoxaemic pigs, visceral protein was conserved, and hindquarter protein breakdown exceeded the increase in liver protein synthesis, resulting in net whole-body protein loss. Endotoxaemia increased hindquarter and visceral glycolysis and branched-chain amino acid transamination. The rate of efflux of glutamine and alanine from the hindquarters was higher than anticipated from protein breakdown, indicating de novo synthesis of these amino acids during endotoxaemia. In addition to the hindquarters, the portal-drained viscera provided substantial gluconeogenic amino acids and lactate to the liver. Although increased liver glutamate release constitutes an important nitrogen-sparing mechanism and carbon skeletons are effectively being cycled in glucose, net body protein is lost through increased ureagenesis during the hyperdynamic stage of sepsis. Specific amino acid requirements may develop in compensated hyperdynamic sepsis that is characterized by maintained organ perfusion and increased substrate utilization at the expense of body protein. PMID- 12546636 TI - ETA receptors mediate vasoconstriction, whereas ETB receptors clear endothelin-1 in the splanchnic and renal circulation of healthy men. AB - The contribution of the endothelin (ET) receptors ET(A) and ET(B) to basal vascular tone and ET-1-induced vasoconstriction in the renal and splanchnic vasculature was investigated in six healthy humans. ET-1 was infused alone and in combination with the selective ET(A) receptor antagonist BQ123 or the selective ET(B) receptor antagonist BQ788 on three different occasions. BQ123 did not affect basal arterial blood pressure, splanchnic vascular resistance (SplVR) or renal vascular resistance (RVR), but inhibited the increase in vascular resistance induced by ET-1 [64+/-18 versus -1+/-7% in SplVR ( P <0.05); 36+/-6 versus 12+/-3% in RVR ( P <0.0001)]. BQ788 increased basal SplVR and RVR [38+/ 16% ( P =0.01) and 21+/-5% ( P <0.0001) respectively], and potentiated the ET-1 induced vasoconstriction. Plasma ET-1 increased more after ET(B) blockade than under control conditions or after ET(A) blockade. These findings suggest that the ET(A) receptor mediates the splanchnic and renal vasoconstriction induced by ET-1 in healthy humans. The ET(B) receptor seems to function as a clearance receptor and may modulate vascular tone by altering the plasma concentration of ET-1. PMID- 12546637 TI - Effects of creatine loading and prolonged creatine supplementation on body composition, fuel selection, sprint and endurance performance in humans. AB - Most research on creatine has focused on short-term creatine loading and its effect on high-intensity performance capacity. Some studies have investigated the effect of prolonged creatine use during strength training. However, studies on the effects of prolonged creatine supplementation are lacking. In the present study, we have assessed the effects of both creatine loading and prolonged supplementation on muscle creatine content, body composition, muscle and whole body oxidative capacity, substrate utilization during submaximal exercise, and on repeated supramaximal sprint, as well as endurance-type time-trial performance on a cycle ergometer. Twenty subjects ingested creatine or a placebo during a 5-day loading period (20 g.day(-1)) after which supplementation was continued for up to 6 weeks (2 g.day(-1)). Creatine loading increased muscle free creatine, creatine phosphate (CrP) and total creatine content ( P <0.05). The subsequent use of a 2 g.day(-1) maintenance dose, as suggested by an American College of Sports Medicine Roundtable, resulted in a decline in both the elevated CrP and total creatine content and maintenance of the free creatine concentration. Both short- and long-term creatine supplementation improved performance during repeated supramaximal sprints on a cycle ergometer. However, whole-body and muscle oxidative capacity, substrate utilization and time-trial performance were not affected. The increase in body mass following creatine loading was maintained after 6 weeks of continued supplementation and accounted for by a corresponding increase in fat-free mass. This study provides definite evidence that prolonged creatine supplementation in humans does not increase muscle or whole-body oxidative capacity and, as such, does not influence substrate utilization or performance during endurance cycling exercise. In addition, our findings suggest that prolonged creatine ingestion induces an increase in fat-free mass. PMID- 12546638 TI - Orthostatic challenge reveals impaired vascular resistance control, but normal venous pooling and capillary filtration in familial dysautonomia. AB - Patients with familial dysautonomia (FD) frequently have profound orthostatic hypotension without compensatory tachycardia. Although the aetiology is presumed to be sympathetic impairment, peripheral vascular responses to orthostasis have not been assessed. The aim of this study was to evaluate the control of vascular responses to postural stress in FD patients. Measurements of heart rate, blood pressure, cardiac stroke volume and cardiac output (CO), by impedance cardiography, and calf-volume changes, by impedance plethysmography, were taken from nine FD patients and 11 control subjects while supine and during head-up tilt. During leg lowering, we also assessed the venoarteriolar reflex by measuring skin red-cell flux. Head-up tilting for 10 min induced sustained decreases in mean arterial pressure in the FD patients, but not in the controls. Total peripheral resistance (TPR, i.e. mean arterial pressure/CO) increased significantly in the controls (39.8+/-6.8%), but not in the FD patients. Calf volume changes during tilting, when normalized for the initial calf volume, did not differ significantly between the patients (4.62+/-1.99 ml.100 ml(-1)) and the controls (3.18+/-0.74 ml.100 ml(-1)). The vasoconstrictor response to limb lowering was present in the patients (47.7+/-9.0% decrease in skin red-cell flux), but was impaired as compared with the controls (80.7+/-3.4%) ( P <0.05). The impaired vasoconstriction during limb lowering and absent increase of TPR during tilting confirm that orthostatic hypotension in FD is due primarily to a lack of sympathetically mediated vasoconstriction without evidence of abnormally large shifts in blood volume towards the legs during orthostasis. This may be due, in part, to a preserved myogenic response to increased vascular pressure in the dependent vascular beds. PMID- 12546639 TI - Polyunsaturated and monounsaturated fatty acids increase neutral lipid accumulation, caspase activation and apoptosis in a neutrophil-like, differentiated HL-60 cell line. AB - We report here that monounsaturated fatty acids and polyunsaturated fatty acids (PUFAs) provoke the accumulation of neutral lipids and apoptosis in retinoic acid treated HL-60 cells in a concentration- and time-dependent manner. The PUFAs (arachidonic acid, docosahexanoic acid and eicosapentaenoic acid) provoked higher levels of HL-60 apoptosis compared with the monounsaturated oleic acid or the saturated palmitic acid. Cell size and granularity were also altered by fatty acid treatment. The PUFA-induced apoptosis was correlated with increased activity of caspase 3 and caspase 9. Lipid peroxidation was also increased in the presence of PUFAs, but was not responsible for activating cell apoptosis. Lipid derived metabolites may be responsible for activation of caspases and induction of cell apoptosis. PMID- 12546640 TI - Role of transforming growth factor beta in conjunctival scarring. AB - Glaucoma is the major cause of irreversible blindness throughout the world. Of all of the treatments that are available at present, the most effective appears to be surgery; however, excessive conjunctival scarring can lead to surgical failure. In the last decade, the introduction of the anti-metabolites mitomycin-C and 5-fluorouracil as anti-scarring treatments have greatly improved the results of glaucoma surgery, but these agents are associated with complications that can potentially result in blindness. A possible target for a more physiological approach to anti-scarring is transforming growth factor beta. This review examines the role of transforming growth factor beta in conjunctival scarring and discusses promising new ways of modifying its activity. PMID- 12546641 TI - Levels of adhesion molecules do not decrease after 3 months of statin therapy in moderate hypercholesterolaemia. AB - Studies in animals and humans indicate a pivotal role for adhesion molecules (AMs) in the pathogenesis of atherosclerosis. Whereas an association between hypercholesterolaemia and AM expression has been suggested, it is unclear whether lowering cholesterol decreases AM expression and release. We compared the effects of a 3-month treatment with standard doses of three different statins (atorvastatin, simvastatin and pravastatin) on plasma levels of circulating AM (cAM) in 75 hypercholesterolaemic patients in a randomized clinical trial. Plasma levels of circulating (c)E-selectin, circulating intercellular adhesion molecule 1 (cICAM-1) and circulating vascular cell adhesion molecule-1 (cVCAM-1) were measured before and after 3 months of therapy. None of the statins lowered plasma cAM levels and pooled analyses of all patients showed a 1.7% [95% confidence interval (CI), -1.4-4.9%] increase in cE-selectin, a 2.1% (95% CI, -0.2-4.4%) increase in cICAM-1, and a 2.7% (95% CI, -0.6-6.1%) increase in cVCAM-1 levels. cAM levels did not decrease, even in patients with a >50% decrease ( n =19) in low-density lipoprotein cholesterol levels. This study provides strong evidence that 3 months of therapy with three different statins does not decrease cAM levels, despite normalization of cholesterol levels, and a minor decrease in C reactive protein levels in patients with moderate hypercholesterolaemia. PMID- 12546642 TI - Testosterone suppression in men with prostate cancer leads to an increase in arterial stiffness and hyperinsulinaemia. AB - The role of androgens in cardiovascular disease is uncertain. We aimed to determine the vascular effects of androgen suppression in men with prostate cancer. Arterial stiffness (or 'compliance') was measured in 16 men (71+/-9 years, mean+/-S.D.) prior to, and 3 months after, complete androgen suppression with gonadotrophin-releasing hormone analogues as treatment for prostate cancer. Fifteen control men (70+/-7 years) also had arterial stiffness studies at baseline and 3 months later. Two measures of arterial stiffness were employed: systemic arterial compliance (SAC) was measured by simultaneous recording of aortic flow and carotid artery pressure ('area method'), and pulse wave velocities (PWVs) were recorded with the 'Complior' system. The 16 cases underwent glucose-tolerance and fasting-lipids tests on both visits. After 3 months of testosterone suppression, there was a significant fall in SAC, which was not seen in the controls [mean change+/-S.E.M., -0.26+/-0.09 a.c.u. (arbitrary compliance unit) in the cases versus +0.06+/-0.11 in the controls; P =0.03). Central, but not peripheral, PWVs tended to increase in the cases (mean change+/-S.E.M. for aorto-femoral PWV, +0.5+/-0.4 m/s for cases versus -0.3+/-0.3 m/s for controls; P =0.08). After testosterone suppression, fasting insulin levels increased from 6.89+/-4.84 m-units/l to 11.34+/-8.16 m-units/l (mean+/ S.D.), total cholesterol increased from 5.32+/-0.77 mmol/l to 5.71+/-0.82 mmol/l and high-density lipoprotein cholesterol increased from 1.05+/-0.24 mmol/l to 1.26+/-0.36 mmol/l; P <0.005 for all. No significant change occurred in body-mass index, serum glucose, low-density lipoprotein cholesterol or triacylglycerol (triglyceride) levels. Our results indicate that loss of androgens in men leads to an increase in aortic stiffness and serum insulin levels, and may therefore adversely affect cardiovascular risk. PMID- 12546643 TI - Biochemical Society Special Lecture. Nitrate- and nitrite-responsive sensors NarX and NarQ of proteobacteria. AB - Nitrate and nitrite are efficient respiratory oxidants for anaerobic growth. In Escherichia coli, the homologous nitrate reductase (Nar) two-component regulatory systems NarX-NarL and NarQ-NarP collaborate to control anaerobic respiratory gene expression in response to nitrate and nitrite. Several other species classified in the gamma and beta subdivisions of the proteobacteria contain only a single Nar two-component regulatory pair. This raises questions concerning the physiology of anaerobic respiration as well as the evolution, function and cross regulation of two-component regulatory systems. Here, I focus on the sensor histidine kinases NarX and NarQ, and present a comparison of the deduced NarX and NarQ primary sequences from a broad sampling of proteobacteria. This comparison defines shared features, including a large central region of unknown function that appears to be unique to this family of sensor kinases. I then consider the phylogenetic distribution of narX and narQ genes in relation to anaerobic respiratory enzyme repertoire and physiological function. One noteworthy observation is that narXL genes are specifically associated with the structural genes for membrane-bound nitrate reductase, narGHJI, whereas organization and linkage of the narQ and narP genes is quite variable. I conclude with some speculative thoughts on the evolutionary and functional divergence of the NarX NarL and NarQ-NarP regulatory systems. Overall, this analysis aims to provide a basis for future hypothesis and experimentation in this area. PMID- 12546644 TI - Morton Medal Lecture. New insights into the roles of phosphoinositides and inositol polyphosphates in yeast. AB - During the past half century, we have progressed from simply viewing myo inositol-containing glycerophospholipids as quantitatively minor membrane constituents to the present, very striking, situation in which more and more important cellular functions are being assigned to a plethora of phosphorylated derivatives of inositol and phosphatidylinositol. Two such examples are discussed briefly: the activation by environmental stresses of the single phosphoinositidase C of yeast, which is related to the phospholipase C delta s of other eukaryotes, and the involvement of PtdIns(3,5) P (2) in endomembrane trafficking. PMID- 12546645 TI - Liverpool: the early years of biochemistry. AB - The first Chair and department of biochemistry in the U.K. were founded at the University of Liverpool in 1902, thanks to a generous donation by William Johnston, a Liverpool shipowner. The first holder of the Johnston Chair, Benjamin Moore, was a dynamic man, who set up an active research centre. In 1906, he and Edward Whitley founded The Bio-Chemical Journal as a private venture, and in 1912, they sold it to the Biochemical Society. Moore also initiated the first Honours School of Biochemistry in the country before moving to London in 1914 and being succeeded by Walter Ramsden. The development of the department was stopped by World War I, and there was little expansion in the 1920s. After Ramsden's retirement in 1931, the third Johnston Professor, Harold Channon, increased staff numbers, ran a successful research school and re-established the Honours course. World War II brought that to an end, and Channon moved into industry. After the war, biochemistry expanded from a niche subject in a small number of British universities into one that was strongly represented in most universities, but the penetration of biochemistry into wide areas of functional biology has blurred conventional subject boundaries, so in many universities (including the University of Liverpool), departments of biochemistry have been incorporated into large more general schools. PMID- 12546646 TI - Signal transduction by vascular endothelial growth factor receptors. AB - Vascular endothelial growth factor (VEGF)/vascular permeability factor is the prototype for a growing family of dimeric growth factors, which exert their effects on vascular and lymphatic endothelial cells, as well as on a wide range of other cell types. Gene targeting shows that most, if not all, of the factors and receptors in this family serve critical functions during vascular development or in adult physiological and pathological angiogenesis. Growing tumours produce VEGF, and many different strategies for inhibiting tumour growth by inhibiting VEGF production are being tested in clinical trials at present. This review focuses on the signal transduction properties of VEGF receptor-1 and VEGF receptor-2. PMID- 12546647 TI - Isolation of novel Src substrates. AB - We have established and used a method to rapidly isolate tyrosine kinase substrates. The method entails inserting mammalian cDNA libraries into phage vectors. Protein production is induced, then plaque proteins are transferred to nitrocellulose and phosphorylated by the kinase of interest. Proof of principle for this technique was established by the isolation of a number of known Src substrates. We also implicated other known proteins as substrates for Src by this approach, and isolated a number of novel genes. Several of these are indeed Src substrates in mammalian cells. We have characterized further one of these novel substrates, Fish, which is a multi-domain adaptor protein. PMID- 12546648 TI - Structure and activation of muscarinic acetylcholine receptors. AB - A homology model of the M(1) muscarinic acetylcholine receptor, based on the X ray structure of bovine rhodopsin, has been used to interpret the results of scanning and point mutagenesis studies on the receptor's transmembrane (TM) domain. Potential intramolecular interactions that are important for the stability of the protein fold have been identified. The residues contributing to the binding site for the antagonist, N -methyl scopolamine, and the agonist, acetylcholine, have been mapped. The positively charged headgroups of these ligands probably bind in a charge-stabilized aromatic cage formed by amino acid side chains in TM helices TM3, TM6 and TM7, while residues in TM4 may participate as part of a peripheral docking site. Closure of the cage around the headgroup of acetylcholine may be part of the mechanism for transducing binding energy into receptor activation, probably by disrupting a set of Van der Waals interactions between residues lying beneath the binding site that help to constrain the receptor to the inactive state, in the absence of agonist. This may trigger the reorganization of a hydrogen-bonding network between highly conserved residues in the core of the receptor, whose integrity is crucial for achievement of the activated state. PMID- 12546649 TI - Agonist binding to peptide hormone receptors. AB - A fundamental issue in molecular pharmacology is to define how agonist-receptor interaction differs from that of antagonist-receptor interaction. The V(1a) vasopressin receptor (V(1a)R) is a member of a family of related G-protein coupled receptors (GPCRs) that are activated by vasopressin, oxytocin (OT) and related peptides. A segment of the N-terminus that was required for agonist binding, but not antagonist binding, was identified by characterizing truncated V(1a)R constructs. Site-directed mutagenesis revealed that a single residue (Arg(46)) was critical for agonist binding and receptor activation. The N terminus of the related OT receptor (OTR) could recover agonist binding in a chimaeric OTR(N)-V(1a)R construct. Furthermore, Arg(34) of the human OTR, which corresponds to Arg(46) of the rat V(1a)R, provided agonist-specific binding epitopes in the OTR, indicating a conserved function of this locus throughout this GPCR subfamily. Mutation of Arg(46) revealed that high-affinity agonist binding had an absolute requirement for arginine at this position. PMID- 12546650 TI - The complex matter of DNA double-strand break detection. AB - To maintain genomic stability, despite constant exposure to agents that damage DNA, eukaryotic cells have developed elaborate and highly conserved pathways of DNA damage sensing, signalling and repair. In this review, we concentrate mainly on what we know about DNA damage sensing with particular reference to Lcd1p, a yeast protein that functions early in DNA damage signalling, and MDC1 (mediator of DNA damage checkpoint 1), a recently identified human protein that may be involved in recruiting the MRE11 complex to radiation-induced nuclear foci. We describe a model for the DNA damage response in which factors are recruited sequentially to sites of DNA damage to form complexes that can amplify the original signal and propagate it to the multitude of response pathways necessary for genome stability. PMID- 12546651 TI - Neural differentiation of mouse embryonic stem cells. AB - Pluripotent embryonic stem cells can give rise to neuroectodermal derivatives in culture. This potential could be harnessed to generate neurons and glia for cell replacement therapies in the central nervous system and for use in drug discovery. However, current methods of neural differentiation are empirical and relatively inefficient. Here, we review these methodologies and present new tools for quantification, analysis and manipulation of embryonic stem cell neural determination. PMID- 12546652 TI - Neuron-glia interactions during axon guidance in Drosophila. AB - Axons navigate to trace stereotypic trajectories over an environment often rich in glial cells. Once axonal trajectories are defined, their structuring proceeds through multiple fasciculation and defasciculation events, to finally establish the mature bundles. Fasciculation and ensheathment also proceed in close association between axons and glial cells, and ultimately require glia. The cross talk between axons and glia during axon guidance is manifested in: (i) axonal fasciculation and bundling, promoted by glia; (ii) growth cone guidance, as glia function as guidepost cells at choice points; (iii) glial migration patterns, which are influenced by neurons; (iv) cell survival control, which constrains position and number of both cell types; and (iv) connectivity, where an axon contacts its final target aided by glial cells. Understanding the reciprocal interactions between neurons and glia during guidance and fasciculation is absolutely necessary to implement repair of axonal trajectories upon damage. Drosophila can be used as a model system for these purposes. PMID- 12546653 TI - Structure and mechanism of the oestrogen receptor. AB - We have determined the structures of the oestrogen receptor ligand-binding domain in complex with a range of ligands and with fragments of co-regulator proteins. These structures provide insights into the structural mechanisms underlying the receptor's complex pharmacological properties and how the conformation of the receptor modulates its ability to recruit co-regulators that are necessary for transcriptional activation. PMID- 12546654 TI - Glucocorticoid suppression of nuclear factor-kappa B: a role for histone modifications. AB - Corticosteroids are by far the most effective treatment for chronic inflammatory diseases such as asthma. Inflammation in asthma is characterized by the increased expression of multiple inflammatory genes, including those that encode cytokines, chemokines, adhesion molecules, and inflammatory enzymes and receptors. Increased expression of inflammatory genes is regulated by pro-inflammatory transcription factors, such as nuclear factor kappa B (NF-kappa B). These bind to, and activate, co-activator molecules that then acetylate core histones resulting in elevated gene transcription. Corticosteroids reverse histone acetylation at the site of inflammatory gene transcription, either by direct binding of the activated glucocorticoid receptor to NF-kappa B-associated co-activators or by recruitment of histone deacetylases to the activated transcription complex. Understanding how corticosteroids work in asthma may help in designing novel corticosteroids with fewer systemic effects, as well as novel anti-inflammatory approaches. PMID- 12546655 TI - Cross-coupling between the oestrogen receptor and phosphoinositide 3-kinase. AB - Gender differences in cardiovascular mortality are well documented and oestrogen replacement therapy in post-menopausal women is associated with improved outcomes from cardiovascular events. Indeed, oestrogen therapy has been shown to restore endothelial function in post-menopausal women and reduce the development of atherosclerotic lesions. Despite extensive evidence on the beneficial effect of oestrogen, relatively little is known regarding the molecular signalling mechanism(s) by which oestrogen exerts some of its vascular effects. While the nuclear function of the oestrogen receptor is clearly established, previous studies regarding the membrane and cytoplasmic actions of oestrogen remain inconclusive. Cross-coupling of the oestrogen receptor to phosphoinositide 3 kinase signalling suggests a potentially critical non-nuclear action of the oestrogen receptor and considerably broadens our understanding of the cellular effects of oestrogen. PMID- 12546656 TI - Route of steroid-activated macromolecules through nuclear pores imaged with atomic force microscopy. AB - In eukaryotic cells, two concentric membranes, the nuclear envelope (NE), separate the nucleus from the cytoplasm. The NE is punctured by nuclear pore complexes (NPCs; molecular mass 120 MDa) that serve as regulated pathways for macromolecules entering and leaving the nuclear compartment. Transport across NPCs occurs through central channels. Such import and export of macromolecules through individual NPCs can be elicited in the Xenopus laevis oocyte by injecting the mineralocorticoid aldosterone and can be visualized with atomic force microscopy. The electrical NE resistance in intact cell nuclei can be measured in parallel. Resistance increases when macromolecules are engaged with the NPC. This article describe six observations made from these experiments and the conclusions that can be drawn from them. (i) A homogeneous population of macromolecules (approx. 100 kDa) attaches to the cytoplasmic face of the NPC 2 min after aldosterone injection. They are most likely to be aldosterone receptors. After a few minutes, they have disappeared. (ii) Large plugs (approx. molecular mass 1 MDa) appear in the central channels 20 min after hormone injection. They are most likely to be ribonucleoproteins exiting the nucleus. (iii) Electrical resistance measurements in isolated nuclei reveal transient electrical NE resistance peaks: an early (2 min) peak and a late (20 min) peak. Electrical peaks reflect macromolecule interaction with the NPC. (iv) Spironolactone blocks both the early and late peaks. This indicates that classic aldosterone receptors are involved in the pregenomic (early) and post-genomic (late) responses. (v) Actinomycin D and, independently, RNase A block the late electrical peak, confirming that plugs are genomic in nature. (vi) Intracellular calcium chelation blocks both early and late electrical peaks. Thus, the release of calcium from internal stores, which is known to be the first intracellular signal in response to aldosterone, is a prerequisite for the late genomic response. PMID- 12546657 TI - Calcium signalling in and around the nuclear envelope. AB - We have compared calcium mobilization by Ins(1,4,5) P(3)(IP(3)), cADP-ribose (cADPR) and nicotinic acid-adenosine dinucleotide phosphate (NAADP) from the envelope of isolated nuclei with the calcium signalling in intact isolated pancreatic acinar cells. Ca(2+) uptake and release were studied with calcium sensitive fluorescent probes. In the present study, we have shown that all calcium messengers induce Ca(2+) release from the nuclear envelope. Pre-treatment of nuclei with thapsigargin completely abolished the responses to the calcium messengers, indicating that Ca(2+) stores in isolated nuclei are thapsigargin sensitive. Using different pharmacological tools, we show that Ca(2+) release from pancreatic nuclei is unlikely to occur from stores other than those with endoplasmic reticulum characteristics. We conclude that all three calcium messengers can release Ca(2+) from pancreatic acinar nuclear stores, as previously shown for IP(3) and cADPR. It would appear that NAADP releases Ca(2+) from the same IP(3)- and cADPR-sensitive stores with endoplasmic reticulum characteristics. PMID- 12546658 TI - Activated M-phase-promoting factor (MPF) is exported from the nucleus of starfish oocytes to increase the sensitivity of the Ins(1,4,5)P3 receptors. AB - Starfish oocytes that are extracted from the ovaries are arrested at the prophase of the first meiotic division. At this stage of maturation, they are characterized by a large nucleus called the germinal vesicle. Meiosis resumption (maturation) can be induced in vitro by adding the hormone 1-methyladenine (1-MA) to the seawater in which the oocytes are suspended. Earlier work in our laboratory had detected Ca(2+) increases in both the cytoplasm and the nucleus of the oocytes approx. 2 min after the 1-MA challenge. The nuclear Ca(2+) increase was found to be essential for the continuation of the meiotic cycle, since the injection of bis-(o-aminophenoxy)ethane- N,N,N',N' -tetra-acetic acid (BAPTA) into the nuclear compartment completely blocked the re-initiation of the cell cycle. We have recently confirmed, using confocal microscopy, that the cytoplasmic and nuclear Ca(2+) pools are regulated independently and that the nuclear envelope in starfish oocytes is not freely permeated by the Ca(2+) wave that sweeps across the nuclear region. Studies by others have shown that the sensitivity of the Ins(1,4,5) P (3) (IP(3)) receptors (IP(3)Rs) to IP(3) increases during oocyte maturation, so that they release progressively more calcium in response to the injection of IP(3), as maturation proceeds. We have now shown that the increased sensitivity of the IP(3)Rs may depend on the activation of the cyclin-dependent kinase, MPF (M-phase-promoting factor) that occurs in the nucleus. MPF does not directly phosphorylate IP(3)Rs but phosphorylates instead the actin-binding protein actin depolymerization factor (ADF)/cofilin. PMID- 12546659 TI - The role of Rap1 in integrin-mediated cell adhesion. AB - Rap1 is a member of the Ras-like small GTPases. Originally the protein was identified in a genome-wide screen for suppressors of Ras transformation, but the mechanism of this reversion remained elusive. We have investigated the signalling function of Rap1. We observed that Rap1 is activated by a large variety of stimuli, including growth factors, neurotransmitters and cytokines. Common second messengers like cAMP, diacylglycerol and calcium are mediators of this activation. These messengers activate guanine nucleotide exchange factors (GEFs), the most notable of which is Epac (exchange protein directly activated by cAMP). However, the downstream effectors of Rap1 are less clear. Although direct connections of Rap1 with the serine/threonine kinases Raf1 and B-raf have been reported, we were unable to find functional evidence for an interaction of endogenous Rap1 signalling with the Raf/extracellular-signal-regulated kinase (ERK) pathway. Instead we observe a clear connection of Rap1 with inside-out signalling to integrins. Indeed, introduction of a constitutively active Rap1 as well as Epac induces integrin-mediated cell adhesion, whereas inhibition of Rap1 signalling by the introduction of Rap1GAP (GTPase-activating protein) inhibits inside-out activation of integrins. More importantly, activation of a G(s) protein-coupled receptor results in integrin-mediated cell adhesion, by a pathway involving Epac and Rap1. From these results, we conclude that one of the functions of receptor-induced Rap1 activation is inside-out regulation of integrins. PMID- 12546660 TI - A-kinase-anchoring proteins and cytoskeletal signalling events. AB - Targeting of protein kinases and phosphatases to the cytoskeleton enhances the regulation of many signalling events. Cytoskeletal signalling complexes facilitate this process by optimizing the relay of messages from membrane receptors to specific sites on the actin cytoskeleton. These signals influence fundamental cell properties such as shape, movement and division. Targeting of the cAMP-dependent kinase (protein kinase A) and other enzymes to this compartment is achieved through interaction with A-kinase-anchoring proteins (AKAPs). The present paper discusses recent progress on dissecting the biological role of WAVE1 (Wiskott-Alrich syndrome protein family verprolin homology protein 1), an AKAP that assembles a cytoskeletal transduction complex in response to signals that emanate from the low-molecular-mass GTPase, Rac. PMID- 12546661 TI - Integrin-protein kinase C relationships. AB - The integrins have an ability to interact with extracellular matrix proteins to confer adhesive and motile properties on cells. The means by which these activities operate and the manner in which they are integrated with cell functions is of particular relevance to many biological processes. In the present paper, the developing understanding of the bi-directional relationship between the protein kinase C family of signal transducers and integrins is discussed. PMID- 12546662 TI - Phospholipase D1 and potential targets of its hydrolysis product, phosphatidic acid. AB - Phospholipase D (PLD) hydrolyses phosphatidylcholine into phosphatidic acid (PA) and choline. Our work aims to understand the properties of PLD1, and to identify downstream targets of PA. In one set of projects, we have focused on membrane targeting mechanisms and have proposed a hierarchy of signals that allows PLD1 to localize to intracellular membranes. These signals involve a functional pleckstrin homology (PH) domain and its fatty acylation on two adjacent cysteine residues. A nearby Phox homology (PX) domain may modulate the function of the fatty acylated PH domain. This complex array of signals is probably necessitated by the targeting of PLD1 to multiple endocytic and secretory membranes under basal and signal-dependent conditions. In another set of projects, we have used chemically synthesized PA coupled to a solid support in order to identify proteins that interact with this phospholipid. Several proteins have emerged from this screen as potential targets. Some (e.g. ADP-ribosylation factor, coatomer beta subunit) are involved in trafficking and their PA affinity can be understood in terms of their regulated cycling on and off membranes during rounds of transport. Others (sphingosine 1-phosphate kinase and PtdIns4 P 5-kinase) are implicated in pathways that also involve PLD activation. Others still are novel proteins (brain-specific neurochondrin) whose affinity for PA may contribute to an understanding of their cellular function. PMID- 12546663 TI - Regulation, secretion and activity of type III-secreted proteins of enterohaemorrhagic Escherichia coli O157. AB - Enterohaemorrhagic Escherichia coli (EHEC) O157:H7 causes gastrointestinal disease with the potential for life-threatening sequelae. Although Shiga-like toxins are responsible for much of the serious pathology in humans, the bacterium also possesses a type III protein secretion system that is responsible for intimate attachment to host intestinal mucosa. This sophisticated interaction requires co-ordination that is governed by environmental and genetic factors. Ongoing research supports the following model for how EHEC enables and controls this process: (i) specific environmental cues that are present in the host result in the expression of a number of adhesins, including fimbriae, which allow the initial binding to the mucosal surface. The same conditions support the expression of the basal type III secretion apparatus; (ii) targeting and assembly of the translocon requires both an mRNA signal and chaperones, with coupled translation and secretion of translocon proteins, EspA, B and D; (iii) opening up of a conduit between the bacterium and host cell releases a cytoplasmic pool of effector proteins. A consequence of this is increased expression of particular effector proteins. Potentially, different proteins could be released into the cell at different times or have activities modulated with time; (iv) intimate contact between the translocated intimin receptor (Tir) and the bacterial surface factor intimin requires translocon expression to be down-regulated and translocon filaments to be lost. Fluorescent protein fusions allow contact-mediated regulation and protein targeting through the type III secretion system to be studied in detail. PMID- 12546664 TI - Yersinia pestis and plague. AB - Yersinia pestis is the aetiological agent of plague, a disease of humans that has potentially devastating consequences. Evidence indicates that Y. pestis evolved from Yersinia pseudotuberculosis, an enteric pathogen that normally causes a relatively mild disease. Although Y. pestis is considered to be an obligate pathogen, the lifestyle of this organism is surprisingly complex. The bacteria are normally transmitted to humans from a flea vector, and Y. pestis has a number of mechanisms which allow survival in the flea. Initially, the bacteria have an intracellular lifestyle in the mammalian host, surviving in macrophages. Later, the bacteria adopt an extracellular lifestyle. These different interactions with different host cell types are regulated by a number of systems, which are not well characterized. The availability of the genome sequence for this pathogen should now allow a systematic dissection of these regulatory systems. PMID- 12546665 TI - Temporal regulation of outer surface proteins of the Lyme-disease spirochaete Borrelia burgdorferi. AB - In the 20 years since the first agent of Lyme disease was discovered, much interest has focused on the possible biological roles of a few outer surface proteins (Osps) in the alternating life cycle that includes ticks and vertebrate hosts. Two major proteins, OspA and OspC, are differentially regulated by the spirochaete Borrelia burgdorferi during the several days when ticks feed. The reciprocal decrease in OspA with the rapid up-regulation of OspC by the spirochaetes when ticks are feeding suggests that OspA aids in spirochaete attachment while OspC assists in the dissemination of spirochaetes from tick to vertebrate. Future experiments in ticks with mutant spirochaetes that lack these proteins should clarify the speculative functions currently given to these proteins. PMID- 12546666 TI - Sense of smell: recognition and transduction of olfactory signals. AB - The primary processes in odour perception, i.e. recognition and transduction of olfactory stimuli, are mediated by the chemosensory olfactory neurons. Interaction of odorous compounds with suitable receptor proteins in the membrane of a subset of cells elicits chemo-electrical transduction pathways, including second messenger cascades and ion channels, that modulate the excitability of the sensory neurons, i.e. converting the chemical stimulus into electrical impulses. The encoded information is conveyed via the axons onto distinct glomeruli in the olfactory bulb. Olfactory sensory cells expressing the same receptor type are segregated spatially in a distinct zone of the nasal epithelium and converge their axons to one or a few distinct glomeruli. The emerging chemotopic maps are considered to be crucial for processing and encoding sensory information of olfactory stimuli. PMID- 12546667 TI - Pheromones, binding proteins and receptor responses in rodents. AB - Their well-developed chemical communication systems make rodents popular in research that aims to understand the connections between genes, hormones and behaviour. Structural identification of several pheromones in mice, rats and hamsters now makes it feasible to employ their synthetic analogues in probing olfactory neurons and in the study of various pheromone-protein interactions in intimate detail. PMID- 12546668 TI - Plant stress signalling: understanding and exploiting plant-plant interactions. AB - When plants are attacked by insects, volatile chemical signals can be released, not only from the damaged parts, but also systemically from other parts of the plant and this continues after cessation of feeding by the insect. These signals are perceived by olfactory sensory mechanisms in both the herbivorous insects and their parasites. Molecular structures involved can be characterized by means of electrophysiological assays, using the insect sensory system linked to chemical analysis. Evidence is mounting that such signals can also affect neighbouring intact plants, which initiate defence by the induction of further signalling systems, such as those that increase parasitoid foraging. Furthermore, insect electrophysiology can be used in the identification of plant compounds having effects on the plants themselves. It has been found recently that certain plants can release stress signals even when undamaged, and that these can cause defence responses in intact plants. These discoveries provide the basis for new crop protection strategies, that are either delivered by genetic modification of plants or by conventionally produced plants to which the signal is externally applied. Delivery can also be made by means of mixed seed strategies in which the provoking and recipient plants are grown together. Related signalling discoveries within the rhizosphere seem set to extend these approaches into new ways of controlling weeds, by exploiting the elusive potential of allelopathy, but through signalling rather than by direct physiological effects. PMID- 12546669 TI - Arthropod semiochemicals: mosquitoes, midges and sealice. AB - Signalling chemicals play an essential role in arthropod life cycles. They provide the means whereby mates, host and oviposition sites are located and recognized. The identification of such semiochemicals (sex pheromones, host location and habitat-related cues) in mosquitoes, midges and sealice of farmed salmon is revealing ways in which such devastating pests may be monitored and controlled by behavioural means using push-pull strategies to manipulate the pests away from hosts and into traps. PMID- 12546670 TI - Grouping of odorant receptors: odour maps in the mammalian olfactory bulb. AB - The olfactory system is unique in that the sensory input is in the form of molecular information carried in a vast variety of odorants. Nearly 1000 types of odorant receptors mediate the initial detection and discrimination of odorants at the molecular-feature level. The discrimination at the molecular level is converted into that at the cellular level (olfactory sensory neurons) by the one sensory neuron-one odorant receptor rule, and then into that at the neuronal circuit level in the olfactory bulb by the specific olfactory axon connectivity pattern. Individual glomeruli in the olfactory bulb represent a single odorant receptor, and the glomerular sheet at the olfactory bulb surface forms odorant receptor maps. This review focuses on the spatial organization of the glomerular sensory map in the olfactory bulb. The analysis using the optical imaging method suggests that odorant receptors having a common molecular-feature receptive site are grouped together and represented by glomeruli that are localized in topographically fixed domains in the olfactory bulb. The domain organization may be a structural unit for the spatial organization of the glomerular sensory map, and might relate to the olfactory submodality. PMID- 12546671 TI - Olfactory adventures of elephantine pheromones. AB - Understanding the linkage between behaviour of mammals in their natural environment and the molecular basis of their sensory modalities presents challenges to biologists. Our olfactory investigations that involve the largest extant land mammal, the elephant, offer some clues of how these events mesh in sequence. Proboscideans have developed a sophisticatedly organized society and they rank with primates and cetaceans with respect to cognitive abilities. Our studies of discrete, quantifiable pheromone-elicited behaviours demonstrate that Asian elephants utilize their olfactory senses during fundamental, life-strategy decisions, including mate choice, female bonding and male hierarchical sorting. How biologically relevant odorants traverse mucous interfaces to interact with cognate odorant receptors remains a basic question in vertebrate olfaction. We have partially tracked the molecular odour reception trail of behaviourally distinct pheromones, ( Z )-7-dodecenyl acetate and frontalin (1,5-dimethyl-6,8 dioxabicyclo[3.2.1]octane), using approaches developed for insect studies and taking advantage of the extensive, highly mucoidal olfactory and vomeronasal systems that permit detailed investigations of pheromone-binding proteins. We have combined studies of quantifiable responses and behaviours with biochemical and biophysical investigations of the properties of protein-ligand complexes, their sequential pathways and associated protein-ligand fluxes. In the delineation of these sequential integrations of behavioural, biochemical and molecular events, we have discovered novel spatial and temporal adaptations in both the main olfactory and vomeronasal systems. PMID- 12546672 TI - Multiple roles of major urinary proteins in the house mouse, Mus domesticus. AB - The urine of the house mouse, Mus domesticus, contains large amounts of proteins that are specifically synthesized in the liver to be secreted in the urine. These proteins, termed major urinary proteins (MUPs), have multiple roles in the communication of information in urine-derived scent marks. They bind low molecular-mass volatile pheromones, and effect their delivery to the scent mark, followed by a slow release that is controlled by the rate of dissociation from the MUPs. However, this family of proteins is extremely polymorphic, more than might be expected for a simple role of ligand binding and release. We have analysed the polymorphism in wild mice, and have now shown that the pattern of MUPs in the urine acts as a type of individuality 'bar code' that signals the identity of the owner of the scent mark. This multiplicity of function, from a generic ligand-binding property to an extremely specific individuality, sets the MUPs apart from other lipocalin family proteins that are involved in chemical signalling. PMID- 12546673 TI - Chemical signalling in mice. AB - Odours play a critical role in the behaviour and physiology of many species. For mice and probably many other species, including humans, an individual's olfactory identity (its odourtype) is coded in part by a pattern of volatile compounds that is regulated by genes in the major histocompatibility complex, a string of linked genes that is intimately involved in immune function. The mouse olfactory system is exquisitely sensitive to minute variations in odourtypes. Layered within these chemical signals of individuality is information on the age and health status of the mouse. In the case of age, it appears that information is coded based on a pattern of volatile metabolites; we do not know how a mouse detects, for example, the presence of a viral infection in volatiles from an infected mouse. This chemical information serves to regulate mate choice and other aspects of social behaviour. PMID- 12546674 TI - Towards an understanding of the pregnancy-blocking urinary chemosignals of mice. AB - Male mouse urine contains a pregnancy-blocking chemosignal that causes pre implantation pregnancy failure in recently mated female mice. However, females are able to recognize the chemosignal of the male with which they mated, preventing it from aborting his own offspring. The individuality of the pregnancy blocking chemosignal is influenced by genes of the major histocompatibility complex (MHC), although the chemical nature of the signal remains unclear. Possible candidates include fragments of MHC proteins, the highly polymorphic major urinary proteins (MUPs) and the profile of low-molecular-mass volatiles, which possess male pheromonal activity in other contexts. A recent study has found a high-molecular-mass fraction of male urine containing MUPs to be ineffective in eliciting pregnancy block. Moreover, both the pregnancy-blocking activity and the individuality of the signal were associated with the low molecular-mass fraction of male urine. PMID- 12546675 TI - The future: putting Humpty-Dumpty together again. AB - Successful biological analysis requires that we understand the functional interactions between key components of cells, organs and systems, and how these interactions change in disease. This information resides neither in the genome nor in the individual proteins that genes encode. It lies at the level of protein interactions within the context of sub-cellular, cellular, tissue, organ and system structures. There is therefore no alternative to copying Nature and computing these interactions to determine the logic of healthy and diseased states. The rapid growth in biological databases, models of cells, tissues and organs, and the development of powerful computing hardware and algorithms have made it possible to explore functionality in a quantitative manner all the way from the level of genes to the physiological function of whole organs and regulatory systems. Systems biology of the 21st century is set to become highly quantitative, and therefore one of the most computer-intensive disciplines. PMID- 12546676 TI - Communication: from neurons to people; from present to future. AB - From its emergence out of organic chemistry and physiology a century ago, the history of biochemistry is one of shifting research agendas. For organic chemists, the questions were those of structure and composition, while for physiologists, they were questions of function. The dynamic biochemistry of the mid-20th century centred on catalysis, energy flow and metabolism. The emergence of molecular biology ('practising biochemistry without a licence') introduced information in place of energy as an organizing cellular principle, but in doing so forgot dynamics. For Crick's Central Dogma, information -- signals -- flowed in one direction only. Now, proteomics is enabling molecular biologists to rediscover biochemistry once more. Signalling -- the processes of communication across space and time -- occurs at all biological levels. I will review them, and their potential future. Will the metaphor of signalling provide a new organizing principle, one that recognizes the essentially interactive nature of information flow within metabolic webs?: I discuss first, the conservation of signalling molecules at the cellular level over evolutionary time; secondly, the supracellular level of physiological signalling in multicellular organisms -- hormones and neurons; thirdly, supraorganismic signalling and communication -- from pheromones to speech; and finally, signalling and reception within and outside the biochemical community - how do we/can we communicate with one another and the rest of the world? PMID- 12546677 TI - AMP-activated protein kinase, super metabolic regulator. AB - The AMP-activated protein kinase (AMPK) is a metabolic-stress-sensing protein kinase that regulates metabolism in response to energy demand and supply by directly phosphorylating rate-limiting enzymes in metabolic pathways as well as controlling gene expression. PMID- 12546678 TI - A molecular approach to the concerted action of kinases involved in energy homoeostasis. AB - One of the most important duties of a cell is energy homoeostasis. Several kinases, including AMP-activated protein kinase (AMPK), creatine kinase and adenylate kinase, are involved in the immediate response to stress, resulting in energy depletion. Here, we present our view of events preceding the downstream processes mediated by AMPK and leading to reduced energy expenditure and increased energy production. Unfortunately, AMPK is very poorly defined at the molecular level. Thus a procedure for production of AMPK in milligram amounts is presented which will greatly facilitate the functional and structural characterization of this protein kinase. PMID- 12546679 TI - Role of the yeast Snf1 protein kinase in invasive growth. AB - The sucrose non-fermenting 1 (Snf1) protein kinase of Saccharomyces cerevisiae is important for transcriptional, metabolic and developmental responses to glucose limitation. Here we discuss the role of the Snf1 kinase in regulating filamentous invasive growth. Haploid invasive growth occurs in response to glucose limitation and requires FLO11, a gene encoding a cell-surface adhesin. Snf1 regulates transcription of FLO11 by antagonizing the function of two repressors, Nrg1 and Nrg2. Snf1 and the Nrg repressors also affect diploid pseudohyphal differentiation, which is a response to nitrogen limitation, suggesting an unexpected signalling role for the Snf1 kinase. PMID- 12546680 TI - Snf1 protein kinase: a key player in the response to cellular stress in yeast. AB - Sucrose non-fermenting 1 (Snf1) protein kinase, a yeast homologue of mammalian AMP-activated protein kinase, plays a main role in transcriptional activation and repression of gene expression. In addition, Snf1 kinase has a broad role in the cellular response to several forms of stress, such as nutrient limitation, salt stress and heat shock. PMID- 12546681 TI - Long-term regulation of AMP-activated protein kinase and acetyl-CoA carboxylase in skeletal muscle. AB - Evidence is accumulating for roles of AMP-activated protein kinase (AMPK) in controlling glucose uptake, fatty acid oxidation and gene expression in skeletal muscle. Relatively little is known, however, about the control of expression of the AMPK subunit isoforms. Marked differences are noted in subunit expression as a function of muscle fibre type. Expression of the gamma3 subunit isoform increases in fast-twitch red fibres of the rat in response to training. All subunit isoforms are expressed to a lesser extent in rats treated with propylthiouracil (PTU; an inhibitor of thyroid hormone synthesis) for 3 weeks compared with rats given excess thyroid hormones for 3 weeks. An approx. 2-fold increase in acetyl-CoA carboxylase was observed in gastrocnemius of hyperthyroid rats compared with experimentally hypothyroid rats. Thyroid state therefore appears to be one important factor controlling expression of these proteins in skeletal muscle. PMID- 12546682 TI - A possible role for AMP-activated protein kinase in exercise-induced glucose utilization: insights from humans and transgenic animals. AB - Exercise-induced glucose uptake in skeletal muscle is mediated by an insulin independent mechanism, but the actual signals to glucose transport in response to muscle contraction have not been identified. The 5'-AMP-activated protein kinase (AMPK) has emerged as a putative mediator of contraction-induced glucose transport, although no conclusive evidence has been provided so far. Recent experiments in AMPK transgenic mice suggest that glucose transport induced by 5 amino-4-imidazolecarboxamide riboside (AICAR) or hypoxia is mediated by AMPK. In contrast, contraction-induced glucose transport in rodent skeletal muscle induced by electrical stimulation in vitro or in situ is not influenced or is only partially reduced by abolishing both or one of the catalytic AMPK subunits. This is compatible with exercise studies done in humans, where no tight correlation is found between AMPK activity and glucose uptake during exercise. Taken together, these results question an essential role of AMPK in exercise-induced glucose uptake and imply that one or more additional pathways are involved in mediating glucose transport in skeletal muscle during exercise. PMID- 12546683 TI - AMP-activated protein kinase regulation and action in skeletal muscle during exercise. AB - Physical exercise increases muscle glucose uptake, enhances insulin sensitivity and leads to fatty acid oxidation in muscle. The AMP-activated protein kinase (AMPK) is an energy-sensing enzyme that is strongly activated during muscle contraction due to acute decreases in ATP/AMP and phosphocreatine/creatine ratios. Accumulating evidence suggests that AMPK plays an important role in mediating these metabolic processes. Furthermore, AMPK has been implicated in regulating gene transcription and therefore may play a role in some of the cellular adaptations to training exercise. There is also evidence that changes in AMPK activity result in altered cellular glycogen content, suggesting that this enzyme regulates glycogen metabolism. Recent studies have shown that the magnitude of AMPK activation and associated metabolic responses are affected by factors such as glycogen content, exercise training and fibre type. In summary, AMPK regulates several metabolic pathways during acute exercise and modifies the expression of many genes involved in the adaptive changes to exercise training. PMID- 12546684 TI - Role of AMP-activated protein kinase in leptin-induced fatty acid oxidation in muscle. AB - Leptin regulates energy homoeostasis through central and peripheral mechanisms. Initial steps in leptin action include signalling through a cytokine-like receptor which activates the JAK/STAT pathway. We investigated whether the metabolic effects of leptin in muscle could be mediated by the AMP-activated protein kinase (AMP kinase). Through studies involving leptin injection intrahypothalamically or intravenously, as well as incubation of soleus muscle or cultured muscle cells with leptin, we determined that leptin stimulates fatty acid oxidation in skeletal muscle by activating AMP kinase. Leptin exerts this effect directly at the level of muscle and also through the hypothalamic sympathetic nervous system, specifically engaging alpha-adrenergic receptors in muscle. This represents a novel and important pathway mediating leptin's metabolic actions. PMID- 12546685 TI - Malonyl-CoA and AMP-activated protein kinase (AMPK): possible links between insulin resistance in muscle and early endothelial cell damage in diabetes. AB - Based on available evidence, we would propose the following. (i) Excesses of glucose and free fatty acids cause insulin resistance in skeletal muscle and damage to the endothelial cell by a similar mechanism. (ii) Key pathogenetic events in this mechanism very likely include increased fatty acid esterification, protein kinase C activation, an increase in oxidative stress (demonstrated to date in endothelium) and alterations in the inhibitor kappa B kinase/nuclear factor kappa B system. (iii) Activation of AMP-activated protein kinase (AMPK) inhibits all of these events and enhances insulin signalling in the endothelial cell. It also enhances insulin action in muscle; however, the mechanism by which it does so has not been well studied. (iv) The reported beneficial effects of exercise and metformin on cardiovascular disease and insulin resistance in humans could be related to the fact that they activate AMPK. (v) The comparative roles of AMPK in regulating metabolism, signalling and gene expression in muscle and endothelial cells warrant further study. PMID- 12546686 TI - AMP-activated protein kinase regulation of fatty acid oxidation in the ischaemic heart. AB - The heart relies predominantly on a balance between fatty acids and glucose to generate its energy supply. There is an important interaction between the metabolic pathways of these two substrates in the heart. When circulating levels of fatty acids are high, fatty acid oxidation can dominate over glucose oxidation as a source of energy through feedback inhibition of the glucose oxidation pathway. Following an ischaemic episode, fatty acid oxidation rates increase further, resulting in an uncoupling between glycolysis and glucose oxidation. This uncoupling results in an increased proton production, which worsens ischaemic damage. Since high rates of fatty acid oxidation can contribute to ischaemic damage by inhibiting glucose oxidation, it is important to maintain proper control of fatty acid oxidation both during and following ischaemia. An important molecule that controls myocardial fatty acid oxidation is malonyl-CoA, which inhibits uptake of fatty acids into the mitochondria. The levels of malonyl CoA in the heart are controlled both by its synthesis and degradation. Three enzymes, namely AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC) and malonyl-CoA decarboxylase (MCD), appear to be extremely important in this process. AMPK causes phosphorylation and inhibition of ACC, which reduces the production of malonyl-CoA. In addition, it is suggested that AMPK also phosphorylates and activates MCD, promoting degradation of malonyl-CoA levels. As a result malonyl-CoA levels can be dramatically altered by activation of AMPK. In ischaemia, AMPK is rapidly activated and inhibits ACC, subsequently decreasing malonyl-CoA levels and increasing fatty acid oxidation rates. The consequence of this is a decrease in glucose oxidation rates. In addition to altering malonyl CoA levels, AMPK can also increase glycolytic rates, resulting in an increased uncoupling of glycolysis from glucose oxidation and an enhanced production of protons and lactate. This decreases cardiac efficiency and contributes to the severity of ischaemic damage. Decreasing the ischaemic-induced activation of AMPK or preventing the downstream decrease in malonyl-CoA levels may be a therapeutic approach to treating ischaemic heart disease. PMID- 12546687 TI - New targets of AMP-activated protein kinase. AB - The discovery of the AMP-activated protein kinase (AMPK) more than a decade ago has shed much light on the cellular response to stresses characterized by a fall in the concentration of ATP and an increase in the AMP/ATP ratio. All conditions known to increase this ratio activate AMPK, whose major role is to act as an emergency signal to conserve ATP. It does so by inhibiting anabolic processes and by activating pathways producing ATP. In recent years, our laboratory has discovered new targets of AMPK. The purpose of this short review is to summarize our contribution to this field. PMID- 12546688 TI - Physiological role of AMP-activated protein kinase (AMPK): insights from knockout mouse models. AB - AMP-activated protein kinase (AMPK) is viewed as a fuel sensor for glucose and lipid metabolism. To understand better the physiological role of the catalytic AMPK subunit isoforms, we generated two knockout mouse models with the alpha1 (AMPK alpha 1(-/-)) and alpha 2 (AMPK alpha 2(-/-)) catalytic subunit genes deleted. No defect in glucose homoeostasis was observed in AMPK alpha 1(-/-) mice. On the other hand, AMPK alpha 2(-/-) mice presented high plasma glucose levels and low plasma insulin concentrations in the fed period and during the glucose tolerance test. Nevertheless, in isolated AMPK alpha 2(-/-) pancreatic islets, glucose-stimulated insulin secretion was not affected. Surprisingly, AMPK alpha 2(-/-) mice were insulin-resistant and had reduced muscle glycogen synthesis as assessed in vivo by the hyperinsulinaemic euglycaemic clamp procedure. Reduction of insulin sensitivity and glycogen synthesis were not dependent on the lack of AMPK in skeletal muscle, since mice expressing a dominant inhibitory mutant of AMPK in skeletal muscle were not affected and since insulin-stimulated glucose transport in incubated muscles in vitro was normal in AMPK alpha 2(-/-) muscles. Furthermore, AMPK alpha 2(-/-) mice have a higher sympathetic tone, as shown by increased catecholamine urinary excretion. Increased adrenergic tone could explain both decreased insulin secretion and insulin resistance observed in vivo in AMPK alpha 2(-/-) mice. We suggest that the alpha2 catalytic subunit of AMPK plays a major role as a fuel sensor by modulating the activity of the autonomous nervous system in vivo. PMID- 12546689 TI - AMP-activated protein kinase and hepatic genes involved in glucose metabolism. AB - Mammalian AMP-activated protein kinase presents strong structural and functional similarities with the yeast sucrose non-fermenting 1 (Snf1) kinase involved in the derepression of glucose-repressed genes. It is now clearly established that AMP-activated protein kinase in the liver decreases glycolytic/lipogenic gene expression as well as genes involved in hepatic glucose production. This is achieved through a decreased transcriptional efficiency of transcription factors such as sterol-regulatory-element-binding protein-1c, carbohydrate-response element-binding protein, hepatocyte nuclear factor 4 alpha or forkhead-related protein. Clearly, the long-term consequences of AMP-activated protein kinase activation have to be taken into account if activators of this enzyme are to be designed as anti-diabetic drugs. PMID- 12546690 TI - AMP-activated protein kinase regulates gene expression by direct phosphorylation of nuclear proteins. AB - One of the primary functions of AMP-activated protein kinase (AMPK) is to regulate the metabolic pathways in response to reduced cellular energy charge. Most of the known targets of the kinase are cytoplasmic enzymes involved in both catabolic and anabolic metabolism. In addition, activation of AMPK in many cells results in changes in the pattern of gene expression. Although some of these effects are undoubtedly secondary responses to modified cellular metabolism, it is possible that in addition to its well-characterized function in the cytoplasm, AMPK also directly phosphorylates and regulates proteins involved in gene transcription. There are now several examples of transcription factors, cofactors and components of the transcriptional core machinery that are directly phosphorylated and regulated by AMPK. Here I review these examples and discuss the significance of AMPK activity in the nucleus. PMID- 12546691 TI - Glycogen storage disease as a unifying mechanism of disease in the PRKAG2 cardiac syndrome. AB - The AMP-activated protein kinase (AMPK) system was first discovered 30 years ago. Since that time, knowledge of the diverse physiological functions of AMPK has grown rapidly and continues to evolve. Most recently, the observation that spontaneously occurring genetic mutations in the gamma regulatory subunits of AMPK give rise to a skeletal and cardiac muscle disease emphasizes the critical importance of AMPK in the maintenance of health and disease. The cardiac phenotype observed in humans harbouring genetic mutations in the gamma 2 regulatory subunit (PRKAG2) of AMPK is consistent with abnormal glycogen accumulation in the heart. The perturbation of AMPK activity induced by genetic mutations in PRKAG2 and the resultant effect on muscle cell glucose metabolism may be relevant to the issue of targeting AMPK in drug development for insulin resistant diabetes mellitus. PMID- 12546692 TI - Identification and characterization of AMPK gamma 3 mutations in the pig. AB - The RN(-) (rendement Napole, French for Napole yield) phenotype is common in Hampshire pigs and is characterized by a 70% increase in glycogen content in skeletal muscle and large effects on meat characteristics (pH, water content, technological yield and lean meat content). The phenotype is controlled by an autosomal dominant allele designated RN (-). The protein kinase AMP-activated gamma 3 subunit gene, PRKAG3, which encodes the gamma 3 isoform of AMP-activated protein kinase (AMPK), was identified as the causative gene for this phenotype by a pure positional cloning approach. There are now several lines of evidence supporting our interpretation that the RN(-) phenotype is caused by a missense mutation (Arg(200)-->Gln) in PRKAG3. Recent data from another group have revealed the presence of a third functional allele at the PRKAG3 locus, probably caused by a Val(199)-->Ile missense mutation. This allele has opposite effects compared with RN, as it is associated with a low glycogen content. We have confirmed the phenotypic effect of this third allele in a meat-quality study of a Hampshire/Landrace intercross. A physiological characterization of RN(-) carriers and normal pigs showed that the RN(-) pigs utilized glycogen during exercise to the same extent as normal pigs and they showed a significantly faster resynthesis of glycogen after exercise. The results strongly suggest that the Arg(200)-->Gln substitution is not associated with a defect in glycogen degradation, but rather with an increased glucose uptake in skeletal muscle. PMID- 12546693 TI - Selective suppression of AMP-activated protein kinase in skeletal muscle: update on 'lazy mice'. AB - AMP-activated protein kinase (AMPK) is becoming recognized as a critical regulator of energy metabolism in cells. Using a mouse model in which we specifically blocked AMPK activity in muscles, we have demonstrated that activation of AMPK is necessary for the effects of 5-aminoimidazole-4-carboxamide riboside ('AICAR') and hypoxia, and is possibly required for a portion of exercise-induced glucose uptake. These same mice could not maintain sufficient glycogen in their skeletal muscle and it was rapidly depleted when the animals were subjected to mild exercise. Using isolated strips, we observed muscle hypertrophy and increased tiredness in the AMPK-deficient muscle. We also performed microarray analysis and showed dramatic changes of transcription profile in muscles of the lazy mice. These could have a significant impact on muscle function and may contribute to the observed phenotype. PMID- 12546694 TI - Role of the Saccharomyces cerevisiae Rad9 protein in sensing and responding to DNA damage. AB - Eukaryotic cells have evolved surveillance mechanisms, known as DNA-damage checkpoints, that sense and respond to genome damage. DNA-damage checkpoint pathways ensure co-ordinated cellular responses to DNA damage, including cell cycle delays and activation of repair mechanisms. RAD9, from Saccharomyces cerevisiae, was the first damage checkpoint gene to be identified, although its biochemical function remained unknown until recently. This review examines briefly work that provides significant insight into how Rad9 activates the checkpoint signalling kinase Rad53. PMID- 12546695 TI - Protein-protein interactions during mammalian DNA single-strand break repair. AB - The genetic stability of living cells is continually threatened by endogenous reactive oxygen species and other genotoxic molecules. Of particular threat are the thousands of single-strand breaks that arise in each cell every day. If left unrepaired, such breaks can give rise to potentially clastogenic or lethal chromosomal double-strand breaks. This article summarizes our current understanding of how mammalian cells detect and repair single strand breaks, and provides insights into novel polypeptide components of this process. PMID- 12546696 TI - The RAD30 cancer susceptibility gene. AB - The human skin cancer-prone disease xeroderma pigmentosum variant (XPV) results from a mutation in RAD30, which encodes the novel lesion bypass DNA polymerase eta. XPV cells are characterized by delayed completion of DNA replication and increased mutagenesis following UV irradiation. In cell-free extracts of XPV lymphoblasts, functional DNA polymerase eta is required for the complete replication of a double-stranded plasmid containing either a single (6-4) photoproduct or a cyclobutane pyrimidine dime(CPD), the major mutagenic UV induced lesion. In cultured XPV cells, replication arrest activates downstream signalling pathways, leading to hyperphosphorylation of the 34-kDa subunit of the trimeric single-stranded DNA-binding protein, RPA (replication protein A). Many of the RAD30 mutations identified in XPV cells result in truncation and inactivation of DNA polymerase eta. To examine whether polymorphisms in the RAD30 gene that result in altered polymerase eta function, rather than enzyme inactivation, might contribute to individual susceptibility to skin cancer, methods to screen for sequence changes in the RAD30 gene in human genomic DNAhave been developed. PMID- 12546697 TI - Role played by BRCA1 in regulating the cellular response to stress. AB - BRCA1 (breast-cancer susceptibility gene 1) is a tumour suppressor gene that is mutated in the germline of women with a genetic predisposition to breast and ovarian cancer. In this review, we examine the role played by BRCA1 in mediating the cellular response to stress. We review the role played by BRCA1 in detecting and signalling the presence of DNA damage, particularly double-strand DNA breaks, and look at the evidence to support a role for BRCA1 in regulating stress response pathways such as the c-Jun N-terminal kinase/stress-activated protein kinase pathway. In addition, we examine the role played by BRCA1 in mediating both cell-cycle arrest and apoptosis following different types of cellular insult, and how this may be modulated by the presence or absence of associated proteins such as p53. Finally, we explore the possibility that many of the functions associated with BRCA1 may be based on transcriptional regulation of key downstream genes that have been implicated in the regulation of these specific cellular pathways. PMID- 12546698 TI - Sister chromatid cohesion and genome stability in vertebrate cells. AB - For successful eukaryotic mitosis, sister chromatid pairs remain linked after replication until their kinetochores have been attached to opposite spindle poles by microtubules. This linkage is broken at the metaphase-anaphase transition and the sisters separate. In budding yeast, this sister chromatid cohesion requires a multi-protein complex called cohesin. A key component of cohesin is Scc1/Mcd1 (Rad21 in fission yeast). Disruption of the chicken orthologue of Scc1 by gene targeting in DT40 cells causes premature sister chromatid separation. Cohesion between sister chromatids is likely to provide a substrate for post-replicative DNA repair by homologous recombination. In keeping with this role of cohesion, Scc1 mutants also show defects in the repair of spontaneous and induced DNA damage. Scc1-deficient cells frequently fail to complete metaphase chromosome alignment and show chromosome segregation defects, suggesting aberrant kinetochore function. Consistent with this, the chromosomal passenger protein, INCENP (inner centromere protein) fails to localize to centromeres. Survivin, another passenger protein and one which interacts with INCENP, also fails to localize to centromeres in Scc1-deficient cells. These results show that cohesin maintains genomic stability by ensuring appropriate DNA repair and equal chromosome segregation at mitosis. PMID- 12546699 TI - Regulation of eukaryotic DNA replication at the initiation step. AB - The studies of cell growth and division have remained at the centre of biomedical research for more than 100 years. The combination of genetic, biochemical, molecular and cell biological techniques recently yielded a burst in what is known of the molecular control of cell growth processes. The initiation of DNA replication is crucial for the stability of the genetic information of a cell. Two factors, Cdc45p (cell division cycle 45p) and DNA polymerase alpha-primase, are necessary in this process. Depending on growth signals, Cdc45p is expressed as a late protein. New phosphorylation-specific antibodies specifically recognize the phosphorylated subunit, p68, of the four subunit DNA polymerase alpha-primase and show that the phosphorylated polypeptide is exclusively nuclear. PMID- 12546700 TI - PI3K-signalling in B- and T-cells: insights from gene-targeted mice. AB - Phosphoinositide 3-kinases (PI3Ks) are important signalling enzymes in most cell types. Recent gene targeting studies have shed light on the importance of this family of lipid kinases in the immune system, and the complex mechanisms by which these kinases are regulated in vivo. We have recently reported a phenotype of mice in which the p110 delta PI3K catalytic subunit was inactivated by point mutation. In the present paper, we compare and contrast the phenotypes of p110 delta mutant mice with those of mice that lack p85 alpha or p110 gamma, and discuss these in the context of PI3K signalling in B- and T-cells. PMID- 12546701 TI - Phosphoinositide 3-kinase gamma: a key modulator in inflammation and allergy. AB - Chronic inflammation and allergy involve the activation of tissue-resident cells and, later on, the invasion of effector cells. We have previously shown that the loss of phosphoinositide 3-kinase (PI3K) gamma impairs chemokine-dependent migration of neutrophils and macrophages both in vitro and in vivo. On the other hand, PI3K gamma is not required either during phagocytic processes or in the activation of bactericidal activities like granule secretion and particle mediated respiratory burst in neutrophils. Tissue mast cells are key regulators in allergy and inflammation and release histamine upon clustering of their IgE receptors. We have demonstrated that murine mast cell responses are exacerbated in vitro and in vivo by autocrine signals, and require functional PI3K gamma. Adenosine, acting through the A(3) adenosine receptor, as well as other agonists of G(alpha i)-coupled receptors, transiently increased PtdIns(3,4,5) P (3) exclusively via PI3K gamma. PI3K gamma-derived PtdIns(3,4,5) P (3) was instrumental for initiation of a sustained influx of external Ca(2+) and degranulation. Mice that lacked PI3K gamma did not form oedema when challenged by passive systemic anaphylaxis. PI3K gamma thus relays inflammatory signals through various GPCRs, and is thus central to mast cell function. Taken together, this suggests that pharmaceutical targeting of PI3K gamma might alleviate inflammation at both early and late stages of the allergic response. PMID- 12546702 TI - Fc gamma RIIB activation leads to inhibition of signalling by independently ligated receptors. AB - The inhibitory IgG receptor, Fc gamma RIIB, blocks signalling by co-aggregated antigen receptors on mature and activated B-cells. Fc gamma RIIB is also expressed by immature B-cells; however, its function on these cells has not been defined. In the present paper, we demonstrate that immature B-cells are highly sensitive to inhibitory signalling mediated by Fc gamma RIIB. Co-aggregation of Fc gamma RIIB with the B-cell antigen receptor (BCR) on immature B-cells leads to near ablation of late phase calcium mobilization. Concomitant with enhanced inhibitory signalling, we found that Src-homology-2-domain-containing inositol 5' phosphatase (SHIP) is expressed at much higher levels in immature B-cells than in mature B-cells. Perhaps most importantly, we report that SHIP activated by BCR-Fc gamma RIIB co-aggregation inhibits independently ligated receptors whose signalling requires PtdIns(3,4,5) P (3). We found that stromal-derived factor 1 (SDF-1)-induced cell migration is impaired by prior activation of Fc gamma RIIB. This inhibition is reduced in SHIP-deficient B-cells. Therefore receptor-mediated signalling responses that are dependent on PtdIns(3,4,5) P (3) are subject to both direct and indirect inhibition by Fc gamma RIIB-activated SHIP. PMID- 12546703 TI - Role of Src homology 2-containing-inositol 5'-phosphatase (SHIP) in mast cells and macrophages. AB - The haemopoietic-restricted Src homology 2-containing inositol 5'-phosphatase (SHIP) acts as a negative regulator of myeloid cell proliferation, survival and end-cell activation. It does so, at least in part, by hydrolysing the phosphoinositide 3-kinase (PI3K)-generated second messenger, PtdIns(3,4,5) P (3) (PI-3,4,5-P(3)) to PtdIns(3,4) P (2). As a result, the myeloid progenitors in SHIP-knockout mice display enhanced survival and proliferation and the mice have increased numbers of neutrophils and monocytes/macrophages. Interestingly, although SHIP is not required for mast cell or macrophage development, it restrains their differentiation since progenitors from SHIP(-/-) mice differentiate into mature mast cells and macrophages significantly faster than their wild-type counterparts. This could suggest that elevated PI-3,4,5-P(3) levels accelerate myeloid differentiation. In bone-marrow-derived mast cells, SHIP prevents degranulation by IgE alone, restrains IgE-antigen-induced degranulation and limits the production of inflammatory cytokines. On the other hand, in peritoneal macrophages, SHIP is a positive regulator of NO production, since SHIP(-/-) peritoneal macrophages produce 5-10-fold less NO than their wild type counterparts, even though they show greater lipopolysaccharide/interferon gamma-induced nuclear factor kappa B activation and more rapid inducible NO synthase (iNOS) generation. This is a result of 10-fold higher levels of arginase I in the SHIP(-/-) macrophages, which redirects the iNOS substrate, L-arginine, from NO to ornithine production. This suggests that the chronically elevated PI 3,4,5-P(3) levels in SHIP(-/-) mice may convert M1 (killing) macrophages, which produce NO to kill micro-organisms and tumour cells, into M2 (healing) macrophages, which produce ornithine to promote host cell growth and fibrosis. PMID- 12546704 TI - Regulation of cell survival and proliferation by the FOXO (Forkhead box, class O) subfamily of Forkhead transcription factors. AB - Recently, the FOXO (Forkhead box, class O) subfamily of Forkhead transcription factors has been identified as direct targets of phosphoinositide 3-kinase mediated signal transduction. The AFX (acute-lymphocytic-leukaemia-1 fused gene from chromosome X), FKHR (Forkhead in rhabdomyosarcoma) and FKHR-L1 (FKHR-like 1) transcription factors are directly phosphorylated by protein kinase B, resulting in nuclear export and inhibition of transcription. This signalling pathway was first identified in the nematode worm Caenorhabditis elegans, where it has a role in regulation of the life span of the organism. Studies have shown that this evolutionarily conserved signalling module has a role in regulation of both cell cycle progression and cell survival in higher eukaryotes. These effects are co ordinated by FOXO-mediated induction of a variety of specific target genes that are only now beginning to be identified. Interestingly, FOXO transcription factors appear to be able to regulate transcription through both DNA-binding dependent and -independent mechanisms. Our understanding of the regulation of FOXO activity, and defining specific transcriptional targets, may provide clues to the molecular mechanisms controlling cell fate decisions to divide, differentiate or die. PMID- 12546705 TI - Biosynthesis of rifamycin SV by Amycolatopsis mediterranei MTCC17 in solid cultures. AB - Studies were performed on the production of rifamycin SV, an ansamycin compound, extensively used for curing tuberculosis, leprosy and several other mycobacterial infections, using a strain of Amycolatopsis mediterranei MTCC17 in solid cultures. Wheat bran was employed as a solid substrate. The culture produced 4 g of rifamycin SV/kg of substrate. Pre-treatment of the substrate with dilute HCl was found to increase the yield of rifamycin SV by 300% (from 4 to 12 g x kg of substrate(-1)). Various process parameters were tested to establish the best conditions for the maximum production of the compound and a initial moisture level of 80%, inoculum size of 40%, initial substrate pH of 7.0, incubation temperature of 26 degrees C and a 7 day fermentation period were found to be optimal. Different solvents were used for the extraction of rifamycin SV from the fermented matter and methanol was found to be most suitable. Under optimized conditions, the yield of rifamycin SV further increased from 12 to 32 g x kg of substrate(-1), showing an 8-fold increase from the initial value. PMID- 12546706 TI - Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA. AB - BACKGROUND: The ability to manipulate the genetic networks underlying the physiological and behavioural repertoires of the adult honeybee worker (Apis mellifera) is likely to deepen our understanding of issues such as learning and memory generation, ageing, and the regulatory anatomy of social systems in proximate as well as evolutionary terms. Here we assess two methods for probing gene function by RNA interference (RNAi) in adult honeybees. RESULTS: The vitellogenin gene was chosen as target because its expression is unlikely to have a phenotypic effect until the adult stage in bees. This allowed us to introduce dsRNA in preblastoderm eggs without affecting gene function during development. Of workers reared from eggs injected with dsRNA derived from a 504 bp stretch of the vitellogenin coding sequence, 15% had strongly reduced levels of vitellogenin mRNA. When dsRNA was introduced by intra-abdominal injection in newly emerged bees, almost all individuals (96%) showed the mutant phenotype. An RNA-fragment with an apparent size similar to the template dsRNA was still present in this group after 15 days. CONCLUSION: Injection of dsRNA in eggs at the preblastoderm stage seems to allow disruption of gene function in all developmental stages. To dissect gene function in the adult stage, the intra-abdominal injection technique seems superior to egg injection as it gives a much higher penetrance, it is much simpler, and it makes it possible to address genes that are also expressed in the embryonic, larval or pupal stages. PMID- 12546707 TI - Stone clearance in lower pole nephrolithiasis after extra corporeal shock wave lithotripsy - the controversy continues. AB - BACKGROUND: To determine factors influencing the clearance of fragments after extra-corporeal shock wave lithotripsy (ESWL) for lower pole calyceal (LPC) stones. METHODS: In the period between July 1998 and Oct 2001, 100 patients with isolated lower polar calyceal calculi or= 14 years, were included in the study. Intravenous urograms (IVU) were reviewed to define the LPC anatomy (width of the infundibulum and pelvicalyceal angle). Study end points i.e. stone free status; number of shock waves used and number of sessions were correlated with variables like LPC anatomy, body mass index and stone size. RESULTS: At three months follow up the clearance for stone size 90 degrees ) had stone clearance of 94 and 100% respectively. For the infundibular width of < 4 mm, the stone clearance was 93% were as for > 4 mm, it was 100%. For body mass index (BMI) less than and > 30 kg/m2, the stone clearance was 92 and 95% respectively. CONCLUSIONS: There is a trend towards more ESWL sessions and shock wave requirement in patients with acute pelvi-calyceal angle and narrow infundibulum but it is not statistically significant. Size ( 15) identified retrospectively for 12 month period (1988 to 1989) within former Yorkshire Health Authority area of the United Kingdom. RESULTS: 125 individuals aged 11-24 years at time of injury were identified. Of these, 109 (87%) were interviewed. Only 20% (95% CI 14-29%) of those interviewed reported no disability. Mean Office of Population Census and Surveys (OPCS) disability score of the remainder was 7.5 (median 5.8, range 0.5 to 19.4). The most commonly encountered areas of disability were behaviour (54%, 95% CI 45 63%), intellectual functioning (39%, 95% CI 31-49%) and locomotion (29%, 95% CI 22-39%). Many respondents reported that their daily lives were adversely affected by their health problems for example, causing problems with work, 54% (95% CI 45 63%), or looking after the home, 28% (95% CI 21-38%). Higher OPCS scores were usually but not always associated with greater impact on daily activities. The burden of caring responsibilities fell largely on informal carers. 51% (95% CI 42 61%) of those interviewed would have liked additional help to cope with their injury and disability. CONCLUSION: The study has revealed significant disability amongst a cohort of young people 5 years post severe injury. Whilst many of these young people were coping well with the consequences of their injuries, others reported continuing problems with the activities of daily life. The factors underpinning the young people's differing experiences and social outcome should be explored. PMID- 12546709 TI - Morphological characterization of the Alpha A- and Alpha B-crystallin double knockout mouse lens. AB - BACKGROUND: One approach to resolving some of the in vivo functions of alpha crystallin is to generate animal models where one or both of the alpha-crystallin gene products have been eliminated. In the single alpha-crystallin knockout mice, the remaining alpha-crystallin may fully or partially compensate for some of the functions of the missing protein, especially in the lens, where both alpha A and alpha B are normally expressed at high levels. The purpose of this study was to characterize gross lenticular morphology in normal mice and mice with the targeted disruption of alpha A- and alpha B-crystallin genes (alpha A/BKO). METHODS: Lenses from 129SvEvTac mice and alpha A/BKO mice were examined by standard scanning electron microscopy and confocal microscopy methodologies. RESULTS: Equatorial and axial (sagittal) dimensions of lenses for alpha A/BKO mice were significantly smaller than age-matched wild type lenses. No posterior sutures or fiber cells extending to the posterior capsule of the lens were found in alpha A/BKO lenses. Ectopical nucleic acid staining was observed in the posterior subcapsular region of 5 wk and anterior subcapsular cortex of 54 wk alpha A/BKO lenses. Gross morphological differences were also observed in the equatorial/bow, posterior and anterior regions of lenses from alpha A/BKO mice as compared to wild mice. CONCLUSION: These results indicated that both alpha A- and alpha B-crystallin are necessary for proper fiber cell formation, and that the absence of alpha-crystallin can lead to cataract formation. PMID- 12546710 TI - Prenatal diagnosis of Wolf-Hirschhorn syndrome (4p-) in association with congenital hypospadias and foot deformity. AB - BACKGROUND: Wolf-Hirschhorn syndrome is caused by distal deletion of the short arm of chromosome 4 (4p-). We report a case in which intrauterine growth restriction, hypospadias and foot deformity were detected by prenatal ultrasound examination at 29 weeks of gestation. CASE PRESENTATION: A 31-year-old gravida 2 partus 1 woman was referred at 29 weeks' gestation with suspicion of intrauterine growth restriction. Sonographic examination revealed deformity of the right lower limb and undescended testes with an irregular distal penis. A cordocentesis was performed and chromosome analysis revealed a 46,XY,del(4)(p14) karyotype. CONCLUSION: The prenatal detection of intrauterine growth restriction, hypospadias and foot deformity should lead doctors to suspect the presence of Wolf-Hirschhorn syndrome. PMID- 12546711 TI - Atrial fibrillation in chronic dialysis patients in the United States: risk factors for hospitalization and mortality. AB - BACKGROUND: The incidence and risk factors for hospitalized atrial fibrillation have not been previously assessed in a national population of dialysis patients. METHODS: We analyzed the United States Renal Data System (USRDS) Dialysis Morbidity and Mortality Study (DMMS) Wave II in a historical cohort study of hospitalized atrial fibrillation. Data from 3374 patients who started dialysis in 1996 with valid follow-up times were available for analysis, censored at the time of renal transplantation and followed until November 2000. Cox Regression analysis was used to model factors associated with time to first hospitalization for atrial fibrillation (ICD9 code 427.31x) adjusted for comorbidities, demographic factors, baseline laboratory values, blood pressures, dialysis modality, and cardioprotective medications. RESULTS: The incidence density of atrial fibrillation was 12.5/1000 person years. Factors associated with atrial fibrillation were older age (> or = 71 years vs. <48 years), extremes (both high and low) of pre-dialysis systolic blood pressure, dialysis modality (hemodialysis vs. peritoneal dialysis), and digoxin use. Baseline use of coumadin was associated with reduced mortality in patients later hospitalized for atrial fibrillation. CONCLUSIONS: Dialysis patients had a high incidence of atrial fibrillation. This risk was largely segregated among those with established risk factors for atrial fibrillation, and hemodialysis patients. Use of coumadin was associated with improved survival among patients later hospitalized for atrial fibrillation. PMID- 12546713 TI - Polymeric immunoglobulin receptor polymorphisms and risk of nasopharyngeal cancer. AB - BACKGROUND: Epstein-Barr virus (EBV) associated nasopharyngeal cancer (NPC) is an important squamous cell cancer endemic in Southeast Asia and the Far East and can be considered a multifactorial genetic disease. This research explores potential associations between nasopharyngeal epithelial EBV receptor and NPC susceptibility. To prove the hypothesis, we evaluated two candidate genes, complement receptor 2 (CR2) and polymeric immunoglobulin receptor (PIGR) by using 4 SNPs, CR2IVS2-848C-->T, PIGRIVS3-156G-->T, PIGR1093G-->A and PIGR1739C-->T, to genotype 175 cases and 317 controls, divided into Thai, Chinese and Thai-Chinese based on their respective ethnic origins. RESULTS: The results obtained indicated that PIGR is an NPC susceptibility gene. The risk association pertaining to each ethnic group was detected for homozygous PIGR1739C with a significant ethnic group adjusted OR (95%CI) of 2.71(1.72-4.23) and p < 0.00001. Haplotype of the two missense PIGR SNPs, 1093G-->A and 1739C-->T, and sequence analyses have confirmed the role of the nucleotide PIGR1739 and excluded possibility of an additional significant nonsynonymous NPC susceptibility SNP. CONCLUSIONS: We present genetic evidence leading to hypothesize a possibility of PIGR to function as the EBV nasopharyngeal epithelium receptor via IgA-EBV complex transcytosis failure. The PIGR1739C-->T is a missense mutation changing alanine to valine near endoproteolytic cleavage site. This variant could alter the efficiency of PIGR to release IgA-EBV complex and consequently increase the susceptibility of populations in endemic areas to develop NPC. PMID- 12546712 TI - Gene trap mutagenesis of hnRNP A2/B1: a cryptic 3' splice site in the neomycin resistance gene allows continued expression of the disrupted cellular gene. AB - BACKGROUND: Tagged sequence mutagenesis is a process for constructing libraries of sequenced insertion mutations in embryonic stem cells that can be transmitted into the mouse germline. To better predict the functional consequences of gene entrapment on cellular gene expression, the present study characterized the effects of a U3Neo gene trap retrovirus inserted into an intron of the hnRNP A2/B1 gene. The mutation was selected for analysis because it occurred in a highly expressed gene and yet did not produce obvious phenotypes following germline transmission. RESULTS: Sequences flanking the integrated gene trap vector in 1B4 cells were used to isolate a full-length cDNA whose predicted amino acid sequence is identical to the human A2 protein at all but one of 341 amino acid residues. hnRNP A2/B1 transcripts extending into the provirus utilize a cryptic 3' splice site located 28 nucleotides downstream of the neomycin phosphotransferase start codon. The inserted Neo sequence and proviral poly(A) site function as an 3' terminal exon that is utilized to produce hnRNP A2/B1-Neo fusion transcripts, or skipped to produce wild-type hnRNP A2/B1 transcripts. This results in only a modest disruption of hnRNPA2/B1 gene expression. CONCLUSIONS: Expression of the occupied hnRNP A2/B1 gene and utilization of the viral poly(A) site are consistent with an exon definition model of pre-mRNA splicing. These results reveal a mechanism by which U3 gene trap vectors can be expressed without disrupting cellular gene expression, thus suggesting ways to improve these vectors for gene trap mutagenesis. PMID- 12546715 TI - Pharmacological actions of Uncaria alkaloids, rhynchophylline and isorhynchophylline. AB - The pharmacological actions of Uncaria alkaloids, rhynchophylline and isorhynchophylline extracted from Uncaria rhynchophylla Miq Jacks were reviewed. The alkaloids mainly act on cardiovascular system and central nervous system including the hypotension, brachycardia, antiarrhythmia, and protection of cerebral ischemia and sedation. The active mechanisms were related to blocking of calcium channel, opening of potassium channel, and regulating of nerve transmitters transport and metabolism, etc. PMID- 12546714 TI - Lack of involvement of known DNA methyltransferases in familial hydatidiform mole implies the involvement of other factors in establishment of imprinting in the human female germline. AB - BACKGROUND: Differential methylation of the two alleles is a hallmark of imprinted genes. Correspondingly, loss of DNA methyltransferase function results in aberrant imprinting and abnormal post-fertilization development. In the mouse, mutations of the oocyte-specific isoform of the DNA methyltransferase Dnmt1 (Dnmt1o) and of the methyltransferase-like Dnmt3L gene result in specific failures of imprint establishment or maintenance, at multiple loci. We have previously shown in humans that an analogous inherited failure to establish imprinting at multiple loci in the female germline underlies a rare phenotype of recurrent hydatidiform mole. RESULTS: We have identified a human homologue of the murine Dnmt1o and assessed its pattern of expression. Human DNMT1o mRNA is detectable in mature oocytes and early fertilized embryos but not in any somatic tissues analysed. The somatic isoform of DNMT1 mRNA, in contrast, is not detectable in human oocytes. In the previously-described family with multi-locus imprinting failure, mutation of DNMT1o and of the other known members of this gene family has been excluded. CONCLUSIONS: Mutation of the known DNMT genes does not underlie familial hydatidiform mole, at least in the family under study. This suggests that trans-acting factors other than the known methyltransferases are required for imprint establishment in humans, a concept that has indirect support from recent biochemical studies of DNMT3L. PMID- 12546716 TI - Enhancement of fibrinolytic activity of bovine aortic endothelial cells by ginsenoside Rb2. AB - AIM: The effect of ginsenoside Rb2 purified from Panax ginseng on fibrinolytic activity of bovine aortic endothelial cells (BAEC) was investigated. METHODS: Cellular plasminogen activator (PA) level of the lysates was measured by the chromogenic substrate S-2403. Fibrin underlay technique was carried out to observe fibrinolysis by growing endothelial cells in the culture medium. Cell viability was then determined by measurement of the activity of mitochondrial dehydrogenase. The ability of Rb2 of potentiating cellular PA activity was investigated by measuring the amounts of PA and PA inhibitor-1 (PAI-1) in the culture medium using zymography and reverse zymography. Changes in the expression of urokinase-type PA (uPA), uPA receptor, and PAI-1 mRNA in BAEC after treatment with Rb2 were analyzed by Northern blot. RESULTS: Rb2 enhanced cellular PA activity in a concentration-and time-dependent manner. Treatment of BAEC with Rb2 10 mg/L for 9 h resulted in a 3.5-fold increase of PA activity without a marked cytotoxic effect, as shown by LDH levels in culture. Increased PA levels caused the increase in surface plasmin levels as observed by fibrin underlay technique. Rb2 greatly or moderately increased the amount of urokinase-type PA (uPA) or its inhibitor (PAI-1), present in the culture medium, whereas saponin did not influence mRNA levels of uPA, its surface receptor, and PAI-1, suggesting that Rb2 may stimulate the secretion of uPA without enhancing its gene expression. The enhancement of PA levels by retinoic acid alone, a stimulator of PA synthesis, was potentiated by the simultaneous addition of ginsenoside Rb2 1 mg/L. Therefore, Rb2 might exert a strong synergism in the synthesis of cellular PA in BAEC. CONCLUSION: Ginsenoside Rb2 enhanced the PA activity levels in BAEC as well as the surface plasmin activity of BAEC. Rb2 may stimulate the secretion of uPA without enhancing the gene expression of uPA, uPA receptor (uPAR), and PAI-1. PMID- 12546717 TI - Structure-activity relationship of alkyl 9-nitrocamptothecin esters. AB - AIM: To study the structure-activity relationship of alkyl 9-nitrocamptothecin esters. METHODS: Two alkyl 9-nitrocamptothecin (9NC) esters 5g and 5h were prepared by esterification reactions of 9NC with valeric anhydride and heptanoic anhydride, respectively. Eight 9NC esters 5a-5h were tested for cytotoxicity against human leukemia cell lines HL-60 and U-937. Flow cytometry analysis was used to identify the cell cycle phase targeted by the esters and quantify the extent of ester-induced cell death (apoptosis). RESULTS: Esters 5b and 5c demonstrated great abilities to inhibit growth of the leukemia cells followed by induction of apoptosis; esters 5a, 5e, and 5g induced slight perturbations in the cell cycle at high concentrations; and esters 5d, 5f, and 5h were completely inactive against the cell lines tested. Thus these esters showed the cell anti proliferative activity in an order of 5b approximately 5c>5a approximately 5e approximately 5g>5d approximately 5f approximately 5h. Esters 5b, 5c, and 5e were tested in vivo against various human carcinomas in nude mice grown as xenografts. Only 5b and 5c showed a significant antitumor activity. Particularly, ester 5b demonstrated an antitumor activity against a broad spectrum of human carcinomas including breast, lung, colon, pancreas, stomach, ovarian, and melanoma, etc. CONCLUSION: These esters act like prodrugs of their parental 9-nitrocamptothecin. High drug doses need to be administered to animals in order to inhibit growth, and induce regression, of human tumor xenografts in nude mice. These compounds may be developed into potent anticancer drugs due to their low toxicity. PMID- 12546718 TI - Induction of hemopoiesis by saenghyuldan, a mixture of Ginseng radix, Paeoniae radix alba, and Hominis placenta extracts. AB - AIM: To examine the efficacy of saenghyuldan and its components, Ginseng Radix, Paeoniae Radix Alba, and Hominis Placenta extracts (SHD, GR, PRA, and HP, respectively) on the hemopoiesis in a myelosuppression model system. METHODS: Susceptibility to cyclophosphamide (CP) and S180 carcinoma was determined in SHD, GR, PRA, and HP-treated mice. Analysis of peripheral blood and bone marrow cells was demonstrated by changes in cell types and histopathologic examination. The expression of cytokine mRNAs involved in hemopoiesis was examined by RT-PCR. RESULTS: SHD and its separated components (GR, HP, and PRA, respectively) significantly increased the survival in CP- and S180-treated mice. The hematology data demonstrated that all the agents augmented monocyte and leucocyte counts in the peripheral blood and increased bone marrow density and the ratio of leukocyte to erythrocyte in the bone marrow. These findings were positively correlated with the up-regulation of cytokine mRNA expression such as granulocyte colony stimulating factor (GM-CSF), erythropoietin (EPO), thrombopoietin (TPO), stem cell factor (SCF), and c-Kit. CONCLUSION: SHD is an effective remedy for the bone marrow failure and myelosuppression occurring during chemotherapy. PMID- 12546719 TI - Anti-inflammatory effects of aqueous extract from Dichroa febrifuga root in rat liver. AB - AIM: To study the anti-inflammatory effects of aqueous extract from Dichroa febrifuga root (AEDF) for suppression in the process of lipopolysaccharide (LPS) induced sepsis in the rat liver. METHODS: The inhibitory effect of AEDF on the alteration of inflammatory proteins was investigated by Western blot and immunohistochemical analysis. RESULTS: Western blot analysis showed that the level of nuclear factor (NF)-kappaBp65 was markedly up-regulated and (I) kappaBalpha was down-regulated by LPS (8 mg/kg) challenge. However, AEDF 100 mg/kg inhibited induction of NF-kappaBp65 and degradation of I-kappaBalpha in the liver of LPS-challenged rats. Immunohistochemical analysis showed that while the expression of the NF-kappaBp65, tumor necrosis factor (TNF)-alpha and inducible nitric oxide synthase (iNOS) tended to increase, that of I-kappaBalpha was decreased in the hepatocytes of rats challenged with LPS. A slight decline of NF kappaBp65, TNF-alpha and iNOS, but an increase of I-kappaBalpha were observed in the hepatocytes of the rats pretreated with AEDF. CONCLUSION: AEDF may act as a therapeutic agent for inflammatory disease through a regulation of inflammation related proteins. PMID- 12546720 TI - Effects of gamichunggantang on hyperlipidemia. AB - AIM: To evaluate the therapeutic effects of gamichunggantang (GCT) on hyperlipidemia through high cholesterol diet model. GCT is an Oriental herbal medication, which has been used for the treatment of fatty liver, hyperlipidemia or alcoholic liver disease in Daejeon University Oriental Hospital, Korea since 1999. METHODS: Rats were fed with high cholesterol diet for 4 weeks and GCT was administrated for 2 weeks from 2 weeks later in experimental days. The levels of serum total cholesterol, HDL-cholesterol, and triglyceride were analyzed every week. Absolute and relative liver weight to body, and histopathological changes were determined at last day. And, lipid metabolism-related gene expressions (ACAT and DGAT) in liver tissue were analyzed by using RT-PCR. RESULTS: In GCT group, TG levels were reduced at 3 and 4 weeks after GCT administration (39.4 %, P < 0.05 and 36.3 %, P < 0.01 respectively). Total cholesterol levels also were reduced at 3 weeks (20.5 %, P < 0.05) and 4 weeks (35.86 %, P < 0.01) after GCT administration, but HDL-cholesterol levels were increased significantly (P < 0.05) at 3 weeks (14.7 %) and 4 weeks (25.5 %) compared with hyperlipidemia induced group without GCT. In the GCT treated group, liver weight was lower and lipid accumulation was decreased in histological finding. ACAT gene expression was suppressed compared with hyperlipidemia-induced group but not DGAT. CONCLUSION: GCT possesses preventive or therapeutic effects on diet-induced hyperlipidemia by inhibiting the intestinal absorption and storage of exogenous and endogenous cholesterol. PMID- 12546721 TI - Cytokine production by human lymphocytes stimulated by a herbal compound containing Bupleurum (KY88 LIVER LIVO). AB - AIM: Compounds containing Bupleurum possess immunomodulating effects. KY88 LIVER LIVO (KY88) is a blend of such compound. The aim of this study is to investigate the effects of KY88 on the production of cytokines by lymphocytes in vitro. METHODS: Seventy Sprague Dawley rats were used of which 40 were orally fed with 4 mg purified KY88 for 35 d. Normal human lymphocytes were isolated and cultured in standard conditions. The culture medium was collected at zero and 72 h after the KY88 treatment. The cytokines, including interleukin-1beta (IL-1beta), IL-2, IL 4, IL-6, tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma, were measured by ELISA kits. RESULTS: TNF-alpha levels in the supernatant of cultured human lymphocytes significantly increased after the treatment of PHA and KY88. The mean levels were (855+/-251), (399+/-145), and (176+/-49) ng/L after the treatment with KY88 at the concentrations of 10, 1 and 0.1 g/mL respectively. However, the level in the control group without specific treatment was only (68+/ 4) ng/L. The difference between KY88 10 g/mL and control groups was significant (P<0.05). All other cytokines did not show significant variations between KY88 and the control groups. KY88 may regulate the immune function through the induction of TNF-alpha expression. PMID- 12546722 TI - Evaluation of herbal formulas by pulse analysis method. AB - AIM: To distinguish the component difference between two similar herbal formulas by pulse analysis method. Many Chinese herbs were found to have specific effects on the Fourier components of the blood pressure pulse; it might infer a specific blood redistribution process on the body and reflect the health conditions of specific organs or tissues. The pulse effect of an herbal formula was similar to the linear combination of all its herbal compositions. METHODS: Two different versions of the herbal formula Liu-Wei-Dihuang were fed to the Wistar rats as a single blind test. The blood pressure pulses on the rat tail artery were recorded and then transformed to the frequency domain by Fourier analyzer. RESULTS: Formula A, Bai-Wei-Dihuang, with two more herbs Cortex Cinnamomi and Radix Aconiti added to Liu-Wei-Dihuang, increased the harmonic proportion of the 1st harmonic (C1) but decreased C4, C5. Formula B is composed in the same way but without Rhizoma Batatatis and Poria cocos; it increased the DC of the pulse spectrum (C0), but decreased C2, C3, C4, C5, C6. CONCLUSION: The component adjustment of an herbal formula could be distinctly and quantitatively detected by pulse analysis method. PMID- 12546723 TI - Ginkgo biloba leaf extract enhances levels of caspase-3 and amyloid precursor protein in normal rat hippocampus. AB - AIM: To explore the effect of Ginkgo biloba extract (GbE) on the levels of caspase-3 and amyloid precursor protein (APP) in normal rats' hippocampus. METHODS: Immunohistochemistry method was used for qualitative analysis of the expressions of caspase-3 and APP, and an image analysis method was used for the quantification of the levels of caspase-3 and APP after GbE was administered to rats of different ages for 14 d. RESULTS: The mean absorbance of staining of caspase-3 and APP was markedly higher in GbE group than that in control groups. The expressions of caspase-3 and APP were intensified in the hippocampus of rats after GbE administration. CONCLUSION: GbE can raise the levels of caspase-3 and APP in the hippocampus of normal rats. PMID- 12546724 TI - Effect of Ginkgo biloba leaf extract on electroencephalography of rat with cerebral ischemia and reperfusion. AB - AIM: To test the effect of Ginkgo biloba leaf extract on electroencephalography (EEG) during cerebral ischemia and reperfusion. METHODS: Based on the quantitative analysis of EEG using the fast Fourier transform (FFT), the effect of Ginkgo biloba extract (GbE) on rat EEG was surveyed in the model of middle cerebral artery (MCA) occlusion and global cerebral ischemia. RESULTS: In the global cerebral ischemia, GbE 8 and 16 mg/kg could accelerate the recovery of EEG after reperfusion, and GbE 4 mg/kg had the same effect but much weaker. In the MCA occlusion model, GbE 16 and 32 mg/kg greatly suppressed the drop of power spectrum of EEG. CONCLUSION: GbE could mitigate the cerebral damage caused by ischemia. PMID- 12546725 TI - (-)-Epigallocatechin-3-gallate enhances anti-tumor effect of cytosine arabinoside on HL-60 cells. AB - AIM: To study the potentiation of anti-tumor effect induced by cytosine arabinoside (AraC) with (-)-epigallocatechin-3-gallate (EGCG). METHODS: Growth curve method and MTT assay were used to measure the cytotoxic effect of AraC alone or in combination with EGCG on HL-60 cells. Flow cytometry was used to study the cell cycle distribution of HL-60 cells. Nullification assay was used to examine whether EGCG would nullify the rescue effect of deoxycytidine (dCdR) to AraC. Western blot analysis was employed to investigate bcl-2 expression. Intracellular Ca2+ assay was evaluated. RESULTS: Inhibition of HL-60 cell proliferation induced by AraC was enhanced by EGCG, with multiplication time prolonging from 48 h to 70 h and growth saturation density decreasing from 5.78 to 5.54. The MTT results indicated that IC50 was decreased from (0.34+/-0.29) micromol/L (AraC alone) to (0.11+/-0.09) micromol/L (P<0.05) (in combination with EGCG). Cell cycle analysis indicated that AraC blocked HL-60 cells in G1 phase, inhibited cells in S phase. EGCG had no effect on cell cycle at the current concentration, but enhanced the cell arrest by AraC. Nullification assay indicated that IC50 was 0.03 micromol/L (AraC alone), increased to 0.02 mmol/L when rescued with dCdR, and finally decreased to 4.8 micromol/L when addition with EGCG. The expression of bcl-2 protein was down-regulated after treatment with AraC in combination with EGCG. The intracellular Ca2+ was increased after treatment by AraC in combination with EGCG. CONCLUSION: The combination with EGCG could enhance the anti-tumor effect of AraC on HL-60 cells. PMID- 12546726 TI - Effect of matrine on cold ischemia and reperfusion injury of sinusoidal endothelial cells in rat orthotopic liver transplantation. AB - AIM: To study the mechanism and prevention of matrine (Mat) on cold ischemia/reperfusion injury of sinusoidal endothelial cells (SEC) in rat orthotopic liver transplantation (OLT). METHODS: One hundred and twenty-six syngeneic SD rats were randomly divided into four groups (n=18): untreated group, 40 mg/kg treated group, 80 mg/kg treated group, and pseudo-treated group. After 5 h of preservation in Ringer's (LR) solution, orthotopic implantation of the donor liver was performed. At 1, 2, and 4 h after reperfusion of the portal vein, 6 rats were killed in each group to collect the serum and the median lobe of liver for assay. RESULTS: The level of hylluronic acid (HA) and intercellular adhesion molecule-1 (ICAM-1) decreased significantly in both treated groups at different times post-transplantation, and their pathological changes of SEC were ameliorated, too. CONCLUSION: Matrine can prevent SEC from ischemia and reperfusion injury in rat orthotopic liver transplantation. PMID- 12546727 TI - Protective effects of xanthones against myocardial ischemia-reperfusion injury in rats. AB - AIM: To investigate the protective effect of xanthones against myocardial ischemia-reperfusion injury in rats. METHODS: Ischemia-reperfusion injury was induced by 20 min of global ischemia and 40 min of reperfusion in isolated rat hearts or 60-min coronary artery occlusion and 180-min reperfusion in vivo, respectively. Heart rate, coronary flow, left ventricular pressure (LVP), and its first derivative (+/- dp/dtmax) were recorded, and the activity of creatine kinase in coronary effluent and malondialdehyde contents in myocardial tissues were measured in vitro. The activity of serum creatine kinase and myocardium infarct size were measured in vivo. RESULTS: Xanthones (90 or 300 microg/L) caused a significant improvement of cardiac function (LVP and +/- dp/dtmax) and a decrease in the release of creatine kinase in coronary effluent as well as the level of malondialdehyde in myocardial tissues. Xanthones (0.5 or 1.0 mg/kg) also markedly decreased infarct size and the release of creatine kinase in vivo. CONCLUSION: Xanthones protect the myocardium against the damages induced by ischemia-reperfusion in rats, and the effect of xanthones may be related to the inhibition of lipid peroxidation. PMID- 12546728 TI - Total coumarins from fruits of Cnidium monnieri inhibit formation and differentiation of multinucleated osteoclasts of rats. AB - AIM: To determine the effects of TCFC (total comarins from the fruits of Cnidium monnieri) on the activity of osteoclasts in vitro. METHODS: Osteoclasts isolated from rat marrow cells were co-cultured with osteoblasts under the 1,25 dihydroxyvitamine D3. The tartrate-resistant acid phosphatase (TRAP) stain was used to identify osteoclast morphology. The activity of TRAP was measured by p nitrophenyl sodium phosphate assay. The resorption pit area on the bone slices formed by osteoclasts was measured by computer image processing. Calcium concentration in the medium of co-culture of bone slices and osteoclasts was determined by atomic absorption spectra. RESULTS: TCFC 2.5-25 mg/L inhibited osteoclast formation and differentiation. TCFC 0.25-25 mg/L inhibited TRAP activity of osteoclasts and TCFC 25 mg/L decreased the TRAP activity by 26.3 % and 24.1 % after 48 h and 72 h, respectively. TCFC 25 mg/L decreased the osteoclastic bone resorption pit area by 25.05 % and Ca2+ release from bone slices by 41.73 %. CONCLUSION: TCFC reduced the bone lose by decreasing the osteoclast formation, its TRAP activity, and osteoclastic bone resorption. PMID- 12546729 TI - Anti-inflammatory and anti-platelet aggregation activity of human placental extract. AB - AIM: To find the anti-inflammatory and anti-platelet aggregatory activity of human placental extract (HPE, Placentrex). METHODS: The HPE was studied for anti inflammatory effect in Wistar rats on carrageenin, serotonin (5-HT), and prostaglandin E1 (PGE1) induced edema in acute model and cotton pellet induced granuloma on sub-acute model. Anti-platelet aggregation was studied against protection of adinosine diphosphate (ADP)-induced aggregation of human platelet through in vitro study. RESULTS: HPE showed positive results both in acute and sub-acute models of inflammation. Highly significant (P<0.01) results were obtained against 5-HT induced acute inflammation and cotton pellet induced sub acute inflammation in comparison with standard (diclofenac sodium) and control (normal saline) drugs. The anti-inflammatory property of HPE in animal model was well supported with clinical study of platelet aggregation. There was highly significant (P<0.01) inhibition of platelet aggregation with HPE at different doses against ADP. CONCLUSION: Our data suggest that human placental extract may be useful in suppressing inflammation and platelet aggregation. PMID- 12546730 TI - [Cloning of genes transactivated by hepatitis B virus X protein]. AB - OBJECTIVE: To construct a subtractive cDNA library of genes transactivated by hepatitis B virus X protein (HBX) using suppression subtractive hybridization (SSH) technique and to clone genes associated with HBX transactivating function. METHODS: The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-X and pcDNA3.1(-) empty vector respectively, then cDNA was synthesized. After restriction enzyme RsaI digestion, a number of small size cDNA was obtained. Then tester cDNA was subdivided into two portions and each was ligated with different cDNA adaptor. After tester cDNA was hybridized with driver cDNA twice and underwent nested polymerase chain reaction (PCR) twice the production was subcloned into T/A plasmid vectors to set up the subtractive cDNA library. Amplification of the library was carried out with E. coli strain JM109, some cDNA was sequenced and analyzed in GenBank with Blast. RESULTS: The subtractive cDNA library of genes transactivated by HBX was constructed. The amplified library contained 85 positive clones, and colony PCR showed that these clones contained 200-1000 bp inserts. 65 clones were analyzed by sequencing and bioinformatics, which suggested nineteen known genes and fifteen genes with unknown function. CONCLUSION: A subtractive cDNA library of genes transactivated by HBX using SSH technique has been constructed successfully, which may bring some new clues for studying the biological functions of HBX and the pathogenesis of hepatoma. PMID- 12546731 TI - [Screening of the genes of hepatitis B virus PreS2 interacting proteins]. AB - OBJECTIVE: To screen and clone the genes of proteins in hepatocytes interacting with hepatitis B virus (HBV) PreS2 by yeast-two hybridization technique. METHODS: The HBV PreS2 gene was amplified by polymerase chain reaction (PCR) and HBV PreS2 bait plasmid was constructed by using yeast-two hybridization system 3, then transformed into yeast AH109, followed by mating with yeast Y187 containing liver cDNA library plasmid in 2 YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-Ade-His) and synthetic dropout nutrient medium (SD/-Trp-Leu-Ade-His) containing X-alpha-gal for selecting positive blue clones, then amplified by PCR, sequenced, and performed bioinformatics analysis. RESULTS: HBV PreS2 gene was cloned successfully and expressed in yeast AH109.Twenty-six positive colonies were selected, among them, twelve containing metallothionein 2A, one cytochrome C oxidase II, two cytochrome P450 subfamily IV4F, two cytochrome c oxidase subunit 4 isoform 1, three albumin (ALB), one Na(+)K(+) transporting ATPase beta-1 polypeptide, two prealbumin, one lectin galactoside-binding subunit, and Two new genes with unknown function. CONCLUSION: Genes of HBV PreS2 interacting proteins have been successfully cloned, which brings some new clues for studying the biological functions of HBV PreS2 and related proteins. PMID- 12546732 TI - [Relationship between the genotypes of hepatitis B virus and the severity of liver diseases]. AB - OBJECTIVE: To investigate the relationship between different genotypes of hepatitis B virus (HBV) and the severity of liver diseases. METHODS: The S nucleotide sequences of HBV strains isolated from plasma samples of 284 patients were detected and compared. Among them, 87 patients were HBV asymptomatic carriers (ASC), 157 chronic hepatitis B (CHB), 22 liver cirrhosis (LC), and 18 hepatocellular carcinoma (HCC). RESULTS: Genotypes B and C were predominant, with a 26.1% proportion and a 63.2% proportion respectively. The percentage of genotypes B and C in patients with ASC, CHB, LC, and HCC were significantly different (x(2)=15.09, P<0.001). Compared with genotype B, genotype C was more common in patients with CHB and HCC (59.6% vs 43.2%, chi(2)=10.87, P<0.001; 7.7% vs 1.4%, x(2)=7.41, P<0.001), but in patients with LC there was no different (7.7% vs 8.1%, chi(2)=1.29, P>0.05). CONCLUSION: This study suggests that genotype B and C are predominant. And genotype C may induce more severe the liver inflammation than genotype B may do. PMID- 12546733 TI - [Modulation and implications in pathogenesis of interferon gamma receptor 1 in patients with chronic hepatitis B]. AB - OBJECTIVE: To study the possible differences in the interferon gamma receptor (IFN-gamma R1) response among a variety of clinical types in patients with chronic hepatitis B (CHB) and implications in pathogenesis. METHODS: The serum level of IFN-gamma and the expression level of IFN-gamma R1 in peripheral leucocytes, from 53 CHB patients, were examined by ELISA and flow cytometry respectively, which were compared with the baseline levels of 15 healthy controls and were performed correlation analysis with alanine aminotransferase (ALT), total bilirubin (TBil) in serum and morphological change in hepatic tissues. RESULTS: The results showed that the level of IFN-gamma R1 (28.89% 11.77%) expressed on the membranes of lymphocytes in CHB patients, which correlated with liver inflammation (r=0.621, P<0.01) and serum TBil level (r=0.575, P<0.01), was much higher than that (9.23% 1.30%) of the healthy controls (Z=3.988, P<0.05), and no obvious difference on the membranes of leucocytes. The serum levels of IFN gamma in patients with cirrhosis and severe hepatitis were higher than those of healthy controls. And the two was no difference from each other, but the standard deviation in each group was relatively large. CONCLUSION: These findings suggest that the IFN-gamma signal transduction pathway is modulated through up-regulating the expression of IFN-gamma R1 on the membranes of lymphocytes, which takes part in the immuno-pathogenesis in CHB patients. PMID- 12546734 TI - [Correlation between serum alpha-fetoprotein variant levels in patients with hepatocellular carcinoma and the presence of alpha-fetoprotein mRNA in the circulating blood]. AB - OBJECTIVE: To investigate the relationship between serum alpha-fetoprotein (AFP) variant levels in patients with hepatocellular carcinoma (HCC) and cancer cells disseminating through blood. METHODS: Serum AFP variant levels were measured by crossed immunoaffino-electrophoresis in the presence of lectin before initial surgical treatment in HCC patients. Circulating tumor cells were simultaneously detected in pre-operative blood samples using reverse transcription-polymerase chain reaction (RT-PCR) for AFP mRNA. RESULTS: Forty-six HCC patients with serum AFP positive were studied. Serum AFP variant level > or 20% was showed in 37 patients, among whom there were 22 (59.5%) showing AFP mRNA positive. In contrast, the positive AFP mRNA expression was only observed in 2 out of 9 patients (22.2%) with AFP variant level<20% (x(2)=4.02, P<0.05). CONCLUSION: In hepatocellular carcinoma patients, increased AFP variant levels are associated with a haematogenous spread of tumor cells. PMID- 12546735 TI - [Relationship between genetic polymorphisms of N-acetyltransferase and susceptibility to hepatocellular carcinoma]. AB - OBJECTIVE: To study the possible relationship between genetic polymorphism of N acetyltransferase 2 (NAT2) and susceptibility to hepatocellular carcinoma. METHODS: Genetic polymorphisms of the four NAT2 genes in 78 patients with hepatocellular carcinoma and 112 healthy controls were analyzed by means of real time fluorescence light-Cycler. The difference in frequencies between the hepatocellular carcinoma patients and the controls were compared. RESULTS: The significant difference in slow acetylation genotype frequency was found between the controls and the hepatocellular carcinoma patients who were smokers (17.9% vs 37.5%, x(2)= 4.67, P<0.05) resulting in increased by 2.76 times the risk for hepatocellular carcinoma, but no evident difference between the controls and hepatocellular carcinoma patients who were non-smokers. CONCLUSION: The smokers with slow acetylation genotype of N-acetyltransferase 2 may be the population with high risk for hepatocellular carcinoma. PMID- 12546737 TI - [Effect of Rhein on the development of hepatic fibrosis in rats]. AB - OBJECTIVE: To investigate the effect of rhein on the development of hepatic fibrosis. METHODS: The animal models were made with carbon tetrachloride (CCl(4)) mixed with vegetable oil (3/2, v/v), which was injected subcutaneously twice a week for 6 weeks, and with 5% ethanol for free drinking water. At the same time, Rhein was administrated at the dose of 25 mg/kg or 100 mg/kg once a day for 6 weeks. The changes of both biochemical markers, such as the levels of alanine aminotransferase (ALT), hyaluronic acid (HA), procollagen type III (PCIII) in serum and SOD, malondialdehyde (MDA) in liver, and related histopathological parametres were determined. RESULTS: Compared with the model group, there were three kinds of changes in the larger quantity of rhein treated group. (1) The levels of ALT, HA, PCIII in serum and MDA in liver homogenate were decreased significantly (from 150 U/L +/- 16 U/L to 78 U/L +/- 18 U/L, 321 microg/L +/- 97 microg/L to 217 microg/L +/- 75 microg/L, 31 microg/L +/- 14 microg/L to 16 microg/L +/- 6 microg/L and 3.67 nmol/mg +/- 0.68 nmol/mg to 1.88 nmol/mg +/- 0.34 nmol/mg, respectively, t > or 2.977, P<0.01). However the level of SOD in liver was increased (from 62.45 NU/mg +/- 8.74 NU/mg to 91.26 NU/mg +/- 14.04 NU/mg, t=4.453, P<0.01). (2) The expressions of transforming growth factor beta 1 (TGF-beta 1) and alpha-smooth muscle actin (alpha-SMA) in liver were markedly reduced (P<0.05 and P<0.01). (3) The collagen staining positive area was decreased and the grade of fibrosis was reduced significantly in liver (P<0.05 and P<0.01). CONCLUSION: Rhein can protect hepatocyte from injury and prevent the progress of hepatic fibrosis in rats, which may associate with that rhein plays a role in antioxidation, anti-inflammation, inhibiting the expression of TGF-beta1 and suppressing the activation of hepatic stellate cells (HSCs). PMID- 12546736 TI - [Study on the influence factors of the serum fibrosis markers]. AB - OBJECTIVE: To analyse the factors which influence the four serum fibrosis markers hyaluronic acid (HA), type III procollagen (PCIII), laminin (LN) and type IV collagen (CIV). METHODS: The levels of serum HA, PCIII, LN and CIV were measured by RIA in 141 patients with chronic hepatitis B (CHB), then the patients were divided into two groups according to the serum fibrosis markers, namely consistent group and inconsistent group. the liver biopsy materials were examined pathomorphologically and liver function was detected by automatic biochemistry analyzer, The interior diameters of the portal vein, the spleen vein and the thickness of the spleen were also measured with ultrasonography. RESULTS: 16 patients (14.16%) whose serum fibrosis markers were inconsistent with histological stage of liver fibrosis were found. Their serum fibrosis markers were not correlated with staging of liver fibrosis (P>0.05), but were positively correlated with inflammation grade (x(2)=12.07, P<0.05), at same time, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase(AST), gamma glutamyltransferase (GGT) and globulin (GLB) decreased obviously, from 89.28 U/L +/- 64.25 U/L to 49.31 U/L +/- 26.75 U/L (t=2.45, P<0.05), 66.10 U/L +/- 42.30 U/L to 40.83 U/L +/- 22.40 U/L (t=2.33, P<0.05), 86.26 U/L +/- 70.36 U/L to 48.99 U/L +/- 29.96 U/L (t=2.08, P<0.05) and 32.13 g/L +/- 5.18 g/L to 28.05 g/L +/- 3.47 g/L (t=3.03, P<0.01) respectively. And the level of albumin (ALB) and the ratio of albumin and globulin (A/G) increased evidently, from 42.34 g/L +/- 4.81 g/L to 46.19 g/L +/- 3.61 g/L (t=3.06, P<0.01) and 1.35 +/- 0.28 to 1.63 +/- 0.26 (t=3.70, P<0.01). But the serum level of alkaline phosphatase (ALP), total bilirubin (TBil), total protein (TP), the width of main portal vein, the width of splenic vein and the thickness of the spleen did not change clearly (P>0.05). CONCLUSION: As diagnostic markers, serum fibrosis markers as well as inflammation grade and liver function should be taken into account. PMID- 12546738 TI - [Adult syphilis leading to severe liver lesion: a case report]. PMID- 12546739 TI - [Gene expression analysis in liver tissue at a single cell level by nested polymerase chain reaction and laser microdissection]. AB - OBJECTIVE: To investigate the measurements of gene expressing at a single hepatocyte level. METHODS: Individual hepatocyte was isolated from cryostat tissue section using laser microdissection technique. To detect the mRNA expressed by single hepatocyte, RNA was extracted, reversely transcribed to cDNA and amplified by nested polymerase chain reaction (PCR). RESULTS: Single cell was microdissected from cryostat tissue using an ultraviolet laser micromanipulator. The RNA could be extracted from the isolated cell(s), and the RT-PCR production could be observed after electrophoresis, whose quantitation was compatible with the number of cells. CONCLUSION: Combining laser microdissection and nested RT PCR can monitor gene expression at a single cell level in vivo. PMID- 12546740 TI - [Mitogenic effects of growth and differentiation factors on rat liver stem cell WB-F344 in vitro]. AB - OBJECTIVE: To explore the relationship between the proliferation, differentiation of rat hepatic stem like cell line WB-F344 and cytokines in vitro. METHODS: (3)H thymidine labelling of new synthesized DNA was used to examine the mitogenic responsiveness of WB-F344 cells to cytokines, western blot was used to study the expression of cytokines receptors on hepatic stem cells, and apoptotic cells were detected by Flow cytometry. RESULTS: WB-F344 cells showed a proliferative response to the cytokines of hepatocyte growth factor (HGF), epidermal growth factor (EGF), fibroblast growth factor (FGF), Insulin at the dose of 80 ng/ml, and the relative cpm values are 982.95, 906.32, 863.98 and 968.67 respectively, while non response to interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF alpha) at the same dose, and an inhibition or apoptosis response to transforming growth factor-beta (TGF-beta) at 80 ng/ml with a 26.89% apoptotic rate. Western blot showed that there were HGF, EGF, FGF, TGF-beta receptors expressed on WB F344 cells. When WB-F344 cells were cultured in the differential system (DMEM, 10% Fcs, HGF 10 to approximately 50 ng/ml, EGF 20 ng/ml, Insulin 1 microg/ml, Dex 1 micromol/L), the cells could differentiated into hepatocytes. In addition, HGF could scattered WB-F344 cells. CONCLUSION: The proliferation and differentiation of liver stem cells are regulated by various cytokines which may play an important role when liver is damaged seriously. PMID- 12546741 TI - [Glycocorticosteroid administration prevents fulminant hepatic failure occurrence in patients with chronic hepatitis B of severe degree]. AB - OBJECTIVE: To prevent chronic severe hepatitis, even more fulminant hepatic failure (FHF) occurrence in patients with chronic hepatitis B of severe degree using steroid. METHODS: 120 patients were randomized into conventional supporting treatment and steroid treatment groups. The latter, 62 patients were given intravenously hydrocortisone sodium succinate at the dose of 150 mg to approximately 200 mg everyday plus support care. RESULTS: The rate of deteriorating to chronic severe hepatitis in steroid treatment group was significantly lower than that of conventional group (22% vs 48%, x(2) =7.60, P<0.01). 53.6% (15/28) patients with chronic severe hepatitis in conventional group died, while only 28.6% (4/14) in steroid treatment group succumbed to terminal liver disease (x(2)=0.02, P>0.05). There was no difference between the two groups regarding to complications incidence: gastrointestinal bleeding and infections except for some controllable serious reverse events, such as candidiasis, diabetes, herpes zoster and pulmonary tuberculosis found in some patients in steroid-treated group. CONCLUSION: These results suggest that steroid administration with improved support care not only is likely to prevent chronic severe hepatitis occurrence in patients with chronic viral hepatitis of severe degree, but also shows some efficacy for FHF, which warrant further investigation. PMID- 12546742 TI - [The detection and significance of interleukin-18 and nitric oxide in serum in mice of experimental acute hepatic injury induced with Concanavalin-A]. PMID- 12546743 TI - [Plasmapheresis: intermediate artificial liver]. PMID- 12546744 TI - [Prevention and treatment on complications of artificial liver support system therapy]. PMID- 12546745 TI - [Application of blood purification technique in artificial liver support therapy]. PMID- 12546746 TI - [Cloning and expression of a new gene JST and it's association with liver cancer]. PMID- 12546747 TI - [Histopathological improvement in chronic hepatitis B patients after treatment of marin solution]. PMID- 12546748 TI - [Fabrication and property analysis of an oligochips for detecting hepatitis B virus mutant resistant to Lamivudine]. PMID- 12546749 TI - [The inhibitory effects on hepatitis B virus replication by transient expression of dominant negative mutants of hepatitis B virus X gene]. PMID- 12546750 TI - [Expression and significance of platelet-derived growth factor-BB in liver tissues of patients with chronic hepatitis B]. PMID- 12546751 TI - [Effects of panax notoginseng saponins on the expression of tumor necrosis factor alpha and secretion phospholipase A2 in rats with liver fibrosis]. PMID- 12546752 TI - [Effects of tyrosine protein kinase inhibitor genistein on the cell cycles of activated hepatic stellate cells]. PMID- 12546753 TI - [The implications of monitoring dynamic viral load level in research and therapy for patients infected with hepatitis B virus]. PMID- 12546754 TI - [CCAAT/enhancer-binding protein family and liver diseases]. PMID- 12546755 TI - [Leptin and liver fibrosis]. PMID- 12546756 TI - Use of the Internet among dermatologists in the United Kingdom, Sweden and Norway. AB - The Internet is increasingly important for doctors in all branches of medicine, but to what extent it is used by dermatologists is not known. In May 2001, we sent a questionnaire by post to 1,291 members of the dermatological societies of the United Kingdom, Sweden, and Norway. Six hundred fifty-three (51%) responded. Excluding those from retired doctors and non-dermatologists, 522 questionnaires were available for analysis. Ninety-five percent of the respondents had access to the Internet, at work (77%) and/or at home (83%). Seventy-nine percent used the world wide web for medical updating and other professional purposes. In a multivariate logistic regression analysis, age and private practice were negative predictors for Internet use (p<0.001). Sixty-two percent found medical databases on the Internet and 25% believed the Internet version of medical journals to be important for their continuing medical education. This may be compared to those who found medical journals on paper(83%), medical meetings(81%), and various forms of contact with peers and colleagues(62-66%) to be so. This study shows that a large proportion of dermatologists, especially younger doctors, use the Internet for medical and educational purposes, but Internet use has not yet replaced traditional ways of obtaining continuing medical education. PMID- 12546757 TI - Anti-inflammatory effects of erythromycin and tetracycline on Propionibacterium acnes induced production of chemotactic factors and reactive oxygen species by human neutrophils. AB - Propionibacterium acnes (P. acnes), an anaerobic pathogen, plays an important role in the pathogenesis of acne and seems to initiate the inflammatory process by producing neutrophil chemotactic factors (NCF). Once neutrophils attracted by bacterial chemoattractants reach the inflamed site, they release inflammatory mediators such as lysosomal enzymes and reactive oxygen species (ROS). Previously, it has been shown that antibiotics may affect acne by means other than their anti-bacterial effects. Thus, we investigated the effect of subminimal inhibitory concentration (sub-MIC) of tetracycline and erythromycin on production of NCF and ROS. NCF was tested in vivo in a mouse model and ROS was estimated on human PMNL in vitro, by nitroblue tetrazolium dye reduction test (NBT) and cytochrome-C reduction test. Tetracycline (CS-T) and Erythromycin (CS-E) treated cultures showed a significant reduction of 35.8% and 58.3% in NCF production respectively, as compared to P. acnes stimulated cultures. Tetracycline and erythromycin at their sub-MIC also significantly inhibited release of ROS from human PMNL. Thus, tetracycline and erythromycin, besides having antibacterial activity, also have an anti-inflammatory action. These antibiotics reduce the capacity of P. acnes to produce NCF, as well decrease its ability to induce ROS from PMNL. PMID- 12546759 TI - Squamous cell carcinoma in solid-organ transplantation. AB - Multiple advances in the field of transplantation over the last several decades have resulted in a significant increase in the number of surgeries performed and in years of life extended. However, with the success of transplant comes the dilemma of potential complications of lifelong immunosuppressive therapy. An enormous increase in skin cancer, especially highly-aggressive squamous cell carcinoma, may affect this special population of patients. This review article discusses the main etiologic theories of squamous cell carcinoma, characteristics of skin cancer in the immunosuppressed patient, differences in skin cancer amongst three types of solid-organ transplantation (kidney, heart, and liver), and established and new treatments. PMID- 12546758 TI - Pediatricians who prescribe clotrimazole-betamethasone diproprionate (Lotrisone) often utilize it in inappropriate settings regardless of their knowledge of the drug's potency. AB - Clotrimazole-betamethasone diproprionate (C-BMV) is a fluorinated, high potency topical steroid that has been formulated with clotrimazole in the brand-named product, Lotrisone. The product is frequently used inappropriately in intertriginous areas, particularly in children. The following evaluates the use of this combination based upon a survey of 106 US-based pediatricians with at least two years post-residency, who attended the 1999 American Academy of Pediatrics. Of pediatricians who prescribe C-BMV, 23% prescribe it for diaper dermatitis. 11% of C-BMV prescriptions exceed the recommended duration of therapy. Only 18% of prescribing pediatricians correctly identify "Lotrisone" as a high potency steroid. There is no significant association between knowledge of C-BMV potency and frequency of use (p>.1). These self-reported data confirm and complement the findings of previous studies that used representative national data to assess the use of C-BMV. Pediatricians continue to utilize C-BMV in inappropriate settings, such as diaper dermatitis, regardless of their knowledge of the agent's potency. Our advice is to refrain from using high-potency steroids, such as C-BMV, in pediatric cases as there are more appropriate, safer alternatives with many fewer side effects. PMID- 12546760 TI - Zoonotic sporothricosis transmitted by cats in Rio de Janeiro, Brazil. A case report. AB - Sporotricosis is a ubiquitous mycosis characterized by nodular lesions of the cutaneous or subcutaneous tissues and adjacent lymphatics that usually suppurate and ulcerate. Secondary spread to the articular surface and bone or dissemination to the central nervous system, genitourinary tract or lungs is also possible. All forms of sporothricosis are caused by a single species, Sporothrix schenkii. In the great majority of cases the fungus gains entrance into the body through trauma to the skin with some kind of plant materials such as thorns or splinters. Zoonotic transmission is also possible and several animals are implicated. This kind of transmission is most frequently a professional hazard of people dealing with animals but in some parts of the world, including Rio de Janeiro city and metropolitan region, an increase in transmission by pet cats has been noted. In these cases the infection may be observed in the family environment, an important epidemiological consideration to clinicians. PMID- 12546761 TI - Short course of clarithromycin in an immunocompetent patient with BCG-induced regional complications. AB - Bacillus Calmette Guerin (BCG) has been used to vaccinate against tuberculosis since 1921. Persistant skin and lymph node lesions yielding Mycobacterium bovis are a rare complication for which there is not standardized treatment. We report an 11 month old child with a progressive cutaneous nodule and lymphadenopathy after BCG vaccination. These rapidly cleared with administration of clarithromycin. PMID- 12546765 TI - Angiolymphoid hyperplasia with eosinophilia. PMID- 12546762 TI - Unusual metastatic spread of a malignant eccrine poroma. AB - Malignant eccrine poroma is a rare skin tumor which commonly presents as verrucous plaques or polypoid growths. We report an 83 year old male who presented with an ulcerating eccrine poroma on the left arm. Though the tumor was excised with wide margins, axillary lymphadenopathy due to regional metastasis developed four months later, followed by lymphangitic cutaneous spread at six months. Though the prognosis for this tumor is better than previously thought, it still must certainly be considered potentially fatal. PMID- 12546764 TI - Pemphigus foliaceous. PMID- 12546763 TI - Acquired reactive perforating collagenosis. PMID- 12546766 TI - Bullous morphea. PMID- 12546767 TI - Linear unilateral basal-cell nevus syndrome with comedones. PMID- 12546768 TI - Klippel-Trenaunay syndrome. PMID- 12546769 TI - Leukemia cutis (chronic lymphocytic leukemia-low grade B cell). PMID- 12546770 TI - Pityriasis rotunda. PMID- 12546771 TI - Striate palmoplantar keratoderma. PMID- 12546772 TI - Atypical crusted scabies in an Iranian man. AB - Crusted scabies has been defined as a scabies-induced psoriasiform dermatosis on the hands and feet with dystrophy of the nails and an erythematous scaling eruption that may be generalized. This case report demonstrates a patient who presented with widespread psoriasiform plaques that were teeming with scabies mites and eggs, but who had absence of nail and palmar involvement. While patients with crusted scabies may have nail dystrophy and involvement of the palms, this involvement is not necessary. PMID- 12546773 TI - Langerhans' cell histiocytosis. AB - Adult onset Langerhans cell histiocytosis is uncommon. A 29 year old man is presented who developed classic cutaneous disease with severe systemic involvement. He responded completely to a two month course of thalidomide. PMID- 12546774 TI - AIDS-associated Kaposi's sarcoma in a heterosexual male--a case report. AB - A 39 year old married heterosexual male presented with extensive purpuric macules, papules, and lichenoid plaques scattered over his skin and mucocutaneous surfaces for one year. The lesions were larger in both his groins, and were associated with lymphedema of his penis and scrotum. His serology for the HIV-1 virus was positive. Biopsies from multiple lesions confirmed the diagnosis of Kaposi's sarcoma. Histochemistry was positive for the HHV-8 virus. He subsequently developed pulmonary and peritoneal effusions and died 3 months later of cardiopulmonary arrest. PMID- 12546775 TI - Multiple nodular lesions of upper limbs: nodular mastocytosis. PMID- 12546776 TI - Massive infestation of cutanea larva migrans. AB - Cutaneous larva migrans is a tropical disease that typically is acquired on exposed surfaces in patients who have been sitting or lying on moist ground or other surfaces which have been contaminated by dog or cat feces. A patient is reported who presented with typical, severely pruritic, migratory plaques after sleeping on a wet, bus station floor. PMID- 12546777 TI - Remissions of psoriasis with excimer laser treatment. PMID- 12546778 TI - The increased competitiveness of Mohs micrographic surgery training. PMID- 12546779 TI - Treatment of cutaneous lupus with Elidel. PMID- 12546780 TI - A new method to clean color transparencies. PMID- 12546782 TI - Phylogenetic analyses of diplomonad genes reveal frequent lateral gene transfers affecting eukaryotes. AB - BACKGROUND: Lateral gene transfer (LGT) is an important evolutionary mechanism among prokaryotes. The situation in eukaryotes is less clear; the human genome sequence failed to give strong support for any recent transfers from prokaryotes to vertebrates, yet a number of LGTs from prokaryotes to protists (unicellular eukaryotes) have been documented. Here, we perform a systematic analysis to investigate the impact of LGT on the evolution of diplomonads, a group of anaerobic protists. RESULTS: Phylogenetic analyses of 15 genes present in the genome of the Atlantic Salmon parasite Spironucleus barkhanus and/or the intestinal parasite Giardia lamblia show that most of these genes originated via LGT. Half of the genes are putatively involved in processes related to an anaerobic lifestyle, and this finding suggests that a common ancestor, which most probably was aerobic, of Spironucleus and Giardia adapted to an anaerobic environment in part by acquiring genes via LGT from prokaryotes. The sources of the transferred diplomonad genes are found among all three domains of life, including other eukaryotes. Many of the phylogenetic reconstructions show eukaryotes emerging in several distinct regions of the tree, strongly suggesting that LGT not only involved diplomonads, but also involved other eukaryotic groups. CONCLUSIONS: Our study shows that LGT is a significant evolutionary mechanism among diplomonads in particular and protists in general. These findings provide insights into the evolution of biochemical pathways in early eukaryote evolution and have important implications for studies of eukaryotic genome evolution and organismal relationships. Furthermore, "fusion" hypotheses for the origin of eukaryotes need to be rigorously reexamined in the light of these results. PMID- 12546781 TI - Genetic affinities of the Andaman Islanders, a vanishing human population. AB - BACKGROUND: The Andaman Islands in the Bay of Bengal are inhabited by hunter gatherers of unknown origin, now on the verge of extinction. The Andamanese and other Asian small-statured peoples, traditionally known as "Negritos," resemble African pygmies. However, it is generally believed that they descend from the early Australo-Melanesian settlers of Southeast Asia and that their resemblance to some Africans is due to adaptation to a similar environment, rather than shared origins. RESULTS: We analyzed mitochondrial DNA (mtDNA) sequences and RFLP polymorphisms, and Y chromosome biallelic markers and microsatellites, in present day Andamanese of the Onge, Jarawa, and Great Andamanese tribes, and of inhabitants of the neighboring Nicobar Islands. We also analyzed mtDNA sequences from Andamanese hair samples collected by an ethnographer during 1906-1908. Living Andamanese exhibit low genetic variability that is consistent with their small population size and reproductive isolation. CONCLUSIONS: Our data indicate that the Andamanese have closer affinities to Asian than to African populations and suggest that they are the descendants of the early Palaeolithic colonizers of Southeast Asia. In contrast, the Nicobarese have genetic affinities to groups widely distributed throughout Asia today, presumably descended from Neolithic agriculturalists. PMID- 12546783 TI - Experimental conversion of liver to pancreas. AB - BACKGROUND: The liver and the pancreas arise from adjacent regions of endoderm in embryonic development. Pdx1 is a key transcription factor that is essential for the development of the pancreas and is not expressed in the liver. The aim of this study was to determine whether a gene overexpression protocol based on Pdx1 would be able to cause conversion of liver to pancreas. RESULTS: We show that a modified form of Pdx1, carrying the VP16 transcriptional activation domain, can cause conversion of liver to pancreas, both in vivo and in vitro. Transgenic Xenopus tadpoles carrying the construct TTR-Xlhbox8-VP16:Elas-GFP were prepared. Xlhbox8 is the Xenopus homolog of Pdx1, the TTR (transthyretin) promoter directs expression to the liver, and the GFP is under the control of an elastase promoter and provides a real-time visible marker of pancreatic differentiation. In the transgenic tadpoles, part or all of the liver is converted to pancreas, containing both exocrine and endocrine cells, while liver differentiation products are lost from the regions converted to pancreas. The timing of events is such that the liver is differentiating by the time Xlhbox8-VP16 is expressed, so we consider this a transdifferentiation event rather than a reprogramming of embryonic development. Furthermore, this same construct will bring about transdifferentiation of human hepatocytes in culture, with formation of both exocrine and endocrine cells. CONCLUSIONS: We consider that the conversion of liver to pancreas could be the basis of a new type of therapy for insulin dependent diabetes. Although expression of the transgene is transient, once the ectopic pancreas is established, it persists thereafter. PMID- 12546784 TI - Loss of self-inhibition is a cellular mechanism for episodic rhythmic behavior. AB - BACKGROUND: Rhythmic motor behaviors can be generated continuously (e.g., breathing) or episodically (e.g., locomotion, swallowing), when short or long bouts of rhythmic activity are interspersed with periods of quiescence. Although the mechanisms of rhythm generation are known in detail in many systems, there is very little understanding of how the episodic nature of rhythmic behavior is produced at the neuronal level. RESULTS: Using a well-established episodic rhythm generating neural circuit controlling molluscan feeding, we demonstrate that quiescence between bouts of activity arises from active, maintained inhibition of an otherwise rhythmically active network. We show that the source of the suppressive drive is within the circuit itself; a single central pattern generator (CPG) interneuron type that fires tonically to inhibit feeding during quiescence. Suppression of the tonic activity of this neuron by food is sufficient to change the network from an inactive to a rhythmically active state, with the cell switching function to fire phasically as part of the food-evoked rhythmogenesis. Furthermore, the absolute level of intrinsic suppressive control is modulated extrinsically by the animal's behavioral state (e.g., hunger/satiety), increasing the probability of episodes of feeding when the animal is hungry. CONCLUSIONS: By utilizing the same intrinsic member of a CPG network in both rhythm-generation and suppression, this system has developed a simple and efficient mechanism for generating a variable level of response to suit the animal's changing behavioral demands. PMID- 12546785 TI - Sperm deliver a new second messenger: NAADP. AB - NAADP is a highly potent mobilizer of Ca(2+), which in turn triggers Ca(2+) induced Ca(2+) release pathways in a wide range of species. Nevertheless, NAADP is not presently classified as a second messenger because it has not been shown to increase in response to a physiological stimulus. We now report a dramatic increase in NAADP during sea urchin egg fertilization that was largely due to production in sperm upon contacting egg jelly. The NAADP bolus plays a physiological role upon delivery to the egg based on its ability to induce a cortical flash, a depolarization-induced activation of L-type Ca(2+) channels. Moreover, the sperm-induced cortical flash was eliminated in eggs desensitized to NAADP. We conclude that an NAADP increase plays a physiologically relevant role during fertilization and provides the first conclusive demonstration that NAADP is a genuine second messenger. PMID- 12546786 TI - Trait-to-gene: a computational method for predicting the function of uncharacterized genes. AB - The function of unknown genes is often inferred from comparisons to well characterized homologs. In this paper, we show that, even if all of the homologs of a gene are unannotated, its function may be deduced through phylogenetic profiling. We have designed a series of algorithms that make functional predictions of genes based on orthology and set theory, but our approach to predicting gene function requires no previous knowledge of homolog function. With this technique, we successfully identified 94% of the clusters of orthologous groups that are known to be involved in flagella development or function. As a test, we removed the function of three putative flagellar genes that had been previously uncharacterized in Bacillus subtilis. We observed a motility phenotype for two of these three genes. Thus, these algorithms allow for high-throughput functional prediction of genes beyond that provided by simple orthology-based annotation endeavors. PMID- 12546787 TI - Dedifferentiation of primary spermatocytes into germ cell tumors in C. elegans lacking the pumilio-like protein PUF-8. AB - PUF proteins are a conserved family of RNA binding proteins that regulate RNA stability and translation by binding to specific sequences in 3'-untranslated regions. Drosophila PUMILIO and C. elegans FBF are essential for self-renewal of germline stem cells, suggesting that a common function of PUF proteins may be to sustain mitotic proliferation of stem cells. Here, we show that PUF-8, the C. elegans PUF most related to PUMILIO, performs a different function in germ cells that have begun meiosis: in primary spermatocytes, puf-8 is required to maintain meiosis and prevent the return to mitosis. Primary spermatocytes lacking PUF-8 complete meiotic prophase but do not undergo normal meiotic divisions. Instead, they dedifferentiate back into mitotically cycling germ cells and form rapidly growing tumors. These findings reveal an unexpected ability for germ cells that have completed meiotic prophase to return to the mitotic cycle, and they support the view that PUF proteins regulate multiple transitions during germline development. PMID- 12546788 TI - Temporal mating isolation driven by a behavioral gene in Drosophila. AB - Speciation is the evolutionary process in which new barriers to gene exchange are created. These barriers may be physical, leading to spatial separation of subpopulations and resulting in allopatric speciation, or they may be temporal, giving rise to allochronic speciation, and may include the time of day or the time of year when mating takes place. Drosophila melanogaster and D. pseudoobscura show different temporal patterns of circadian locomotor activity that are determined by the circadian clock gene period (per). Genes that control aspects of behavior that might be relevant to courtship and mating, such as locomotor patterns, become obvious candidates for involvement in the speciation process. However, evidence for the role of individual genes in the mechanism of mate choice has proved elusive. We have used transgenic flies carrying the natural per genes from these two Drosophila species to reveal that per has the potential to provide the permissive conditions for speciation, by affecting mate choice through a mechanism involving the species-specific timing of mating behavior. PMID- 12546789 TI - Prediction precedes control in motor learning. AB - Skilled motor behavior relies on the brain learning both to control the body and predict the consequences of this control. Prediction turns motor commands into expected sensory consequences, whereas control turns desired consequences into motor commands. To capture this symmetry, the neural processes underlying prediction and control are termed the forward and inverse internal models, respectively. Here, we investigate how these two fundamental processes are related during motor learning. We used an object manipulation task in which subjects learned to move a hand-held object with novel dynamic properties along a prescribed path. We independently and simultaneously measured subjects' ability to control their actions and to predict their consequences. We found different time courses for predictor and controller learning, with prediction being learned far more rapidly than control. In early stages of manipulating the object, subjects could predict the consequences of their actions, as measured by the grip force they used to grasp the object, but could not generate appropriate actions for control, as measured by their hand trajectory. As predicted by several recent theoretical models of sensorimotor control, our results indicate that people can learn to predict the consequences of their actions before they can learn to control their actions. PMID- 12546790 TI - Regulation of integrin growth factor interactions in oligodendrocytes by lipid raft microdomains. AB - Individual growth factors can regulate multiple aspects of behavior within a single cell during differentiation, with each signaling pathway controlled independently and also responsive to other receptors such as cell surface integrins. The mechanisms by which this is achieved remain poorly understood. Here we use myelin-forming oligodendrocytes and their precursors to examine the role of lipid rafts, cholesterol and sphingolipid-rich microdomains of the cell membrane implicated in cell signaling. In these cells, the growth factor PDGF has sequential and independent roles in proliferation and survival. We show that the oligodendrocyte PDGFalpha receptor becomes sequestered in a raft compartment at the developmental stage when PDGF ceases to promote proliferation, but is now required for survival. We also show that laminin-2, which is expressed on axons in the CNS and which provides a target-dependent signal for oligodendrocyte survival by amplification of PDGFalphaR signaling, induces clustering of the laminin binding integrin alpha6beta1 with the PDGFalphaR-containing lipid raft domains. This extracellular matrix-induced colocalization of integrin and growth factor receptor generates a signaling environment within the raft for survival promoting PI3K/Akt activity. These results demonstrate novel signaling roles for lipid rafts that ensure the separation and amplification of growth factor signaling pathways during development. PMID- 12546791 TI - The t-SNARE syntaxin 4 is regulated during macrophage activation to function in membrane traffic and cytokine secretion. AB - Activation of macrophages with lipopolysaccharide (LPS) induces the rapid synthesis and secretion of proinflammatory cytokines, such as tumor necrosis factor (TNFalpha), for priming the immune response. TNFalpha plays a key role in inflammatory disease; yet, little is known of the intracellular trafficking events leading to its secretion. In order to identify molecules involved in this secretory pathway, we asked whether any of the known trafficking proteins are regulated by LPS. We found that the levels of SNARE proteins were rapidly and significantly up- or downregulated during macrophage activation. A subset of t SNAREs (Syntaxin 4/SNAP23/Munc18c) known to control regulated exocytosis in other cell types was substantially increased by LPS in a temporal pattern coinciding with peak TNFalpha secretion. Syntaxin 4 formed a complex with Munc18c at the cell surface of macrophages. Functional studies involving the introduction of Syntaxin 4 cDNA or peptides into macrophages implicate this t-SNARE in a rate limiting step of TNFalpha secretion and in membrane ruffling during macrophage activation. We conclude that, in macrophages, SNAREs are regulated in order to accommodate the rapid onset of cytokine secretion and for membrane traffic associated with the phenotypic changes of immune activation. This represents a novel regulatory role for SNAREs in regulated secretion and in macrophage mediated host defense. PMID- 12546792 TI - Single mRNA molecules demonstrate probabilistic movement in living mammalian cells. AB - Cytoplasmic mRNA movements ultimately determine the spatial distribution of protein synthesis. Although some mRNAs are compartmentalized in cytoplasmic regions, most mRNAs, such as housekeeping mRNAs or the poly-adenylated mRNA population, are believed to be distributed throughout the cytoplasm. The general mechanism by which all mRNAs may move, and how this may be related to localization, is unknown. Here, we report a method to visualize single mRNA molecules in living mammalian cells, and we report that, regardless of any specific cytoplasmic distribution, individual mRNA molecules exhibit rapid and directional movements on microtubules. Importantly, the beta-actin mRNA zipcode increased both the frequency and length of these movements, providing a common mechanistic basis for both localized and nonlocalized mRNAs. Disruption of the cytoskeleton with drugs showed that microtubules and microfilaments are involved in the types of mRNA movements we have observed, which included complete immobility and corralled and nonrestricted diffusion. Individual mRNA molecules switched frequently among these movements, suggesting that mRNAs undergo continuous cycles of anchoring, diffusion, and active transport. PMID- 12546793 TI - Mitotic hyperphosphorylation of the fission yeast SIN scaffold protein cdc11p is regulated by the protein kinase cdc7p. AB - The fission yeast septation initiation network (SIN) triggers the onset of septum formation and cytokinesis. SIN proteins signal from the spindle pole body (SPB), to which they bind in a cell cycle-dependent manner, via the scaffold proteins sid4p and cdc11p. cdc11p becomes hyperphosphorylated during anaphase, when the SIN is active. We have investigated the phosphorylation state of cdc11p during mitosis in various mutant backgrounds. We show that association of cdc11p with the spindle pole body is required for its phosphorylation and that ectopic activation of the SIN results in hyperphosphorylation of cdc11p. We demonstrate that mitotic hyperphosphorylation of cdc11p requires the activity of cdc7p and that its dephosphorylation at the end of mitosis requires PP2A-par1p. Furthermore, spindle checkpoint arrest prevents cdc11p hyperphosphorylation. Finally, we show that the septation inhibitor byr4p interacts preferentially with hypophosphorylated cdc11p. We conclude that cdc11p hyperphosphorylation correlates with activation of the SIN and that this may be mediated primarily by cdc7p in vivo. PMID- 12546794 TI - Evolution of eukaryotic cell cycle regulation: stepwise addition of regulatory kinases and late advent of the CDKs. AB - Protein kinases regulate a number of critical events in mitosis and meiosis. A study of the evolution of kinases involved in cell cycle control (CCC) might shed light on the evolution of the eukaryotic cell cycle. In particular, applying quantitative phylogenetic methods to key CCC kinases could provide information on the relative timing of gene duplication events. To investigate the evolution of CCC kinases, we constructed phylogenetic trees for the CDC28 family and performed statistical tests of the tree topology. This family includes the cyclin-dependent kinases (CDKs), which are key regulators of the eukaryotic cell cycle, as well as other CCC kinases. We found that CDKs and, in particular, the principal cell cycle regulator Cdc28p, branch off the phylogenetic tree at a late stage, after several other kinases involved in either mitosis or meiosis regulation. On the basis of this tree topology, it is proposed that, at early stages of evolution, the eukaryotic cell cycle was not controlled by CDKs and that only a subset of extant kinases, notably the DNA damage checkpoint kinase Chk1p, were in place. During subsequent evolution, a series of duplications of kinase genes occurred, gradually adding more kinases to the CCC system, the CDKs being among the last major additions. PMID- 12546795 TI - Vienna focus for Austrian ambitions. AB - The Austrian Academy of Sciences has taken the bold step of closing a successful institute in Salzburg and opening two new ones in Vienna with the aim of creating an internationally attractive critical mass of biological research activity in the capital. Graham Tebb reports. PMID- 12546796 TI - Insider looking out. AB - Josef Penninger, the new director of the planned Institute of Molecular Biotechnology in Vienna, tells Graham Tebb of hurdles but high hopes he has for shaping a new kind of institution in Austria. PMID- 12546797 TI - German gloom on budget prospects. AB - Thousands of German researchers including five Nobel laureates have signed an open letter of protest against the proposed freeze of research funding. Michael Gross reports. PMID- 12546798 TI - Islander's ancient links. AB - The power of genetic analyses to study relationships between human populations is growing rapidly but there are some worries about the use of historically collected human material. Nigel Williams reports on a new study showing the value of such approaches. PMID- 12546799 TI - One wolf wonder. PMID- 12546800 TI - Self-replication. PMID- 12546801 TI - Bacterial chemotaxis. PMID- 12546802 TI - Insulin receptor-like proteins in Tetrahymena thermophila ciliary membranes. PMID- 12546803 TI - Gene transfer: gene swapping craze reaches eukaryotes. AB - Recent studies have provided evidence for gene transfers from prokaryotes to eukaryotes and between eukaryotes. The mechanisms and frequencies of these transfers remain the subject of speculation, but the findings provide ample reason to seriously consider interspecies gene transfer as an important evolutionary process in eukaryotes. PMID- 12546804 TI - Tubulin superfamily: giving birth to triplets. AB - Two new studies show that e tubulin is required for centriole/basal body duplication in both Chlamydomonas and Paramecium, adding to the list of new tubulin family members specifically involved in forming the centriole triplet microtubules. The function of these triplets, and the precise role of e tubulin in triplet formation, remains unclear. PMID- 12546805 TI - Gliding motility: the molecules behind the motion. AB - In apicomplexan parasites, gliding motility and host cell invasion are driven by an actomyosin-based system. Recent studies have characterized several components of the gliding motility apparatus and have provided new insight into the molecular architecture of this locomotory system. PMID- 12546806 TI - Synaptogenesis: Wnt and TGF-beta take centre stage. AB - The formation of functional synapses requires precise coordination between neurons and their synaptic targets. Recent studies have identified two signaling molecules, Wnt and TGF-beta, which are required for formation and growth of the neuromuscular synapse. PMID- 12546807 TI - X-chromosome inactivation: X marks the spot for BRCA1. AB - X-chromosome inactivation equalizes the dosage of X-linked genes in XX females with that in XY males. Recent findings reveal that the BRCA1 breast cancer susceptibility gene has an important function in this epigenetic phenomenon. PMID- 12546808 TI - Prefrontal cortex: procedural sequence learning and awareness. AB - Activation of the prefrontal cortex has been linked to awareness during sequence learning tasks. A recent study, however, finds activation of the prefrontal cortex during such tasks regardless of awareness. So what is the neurophysiological basis of awareness, and what is the role of the prefrontal cortex in sequence learning? PMID- 12546809 TI - Genome evolution: recombination speeds up adaptive evolution. AB - Mutual interference among linked genetic sites subject to selection may reduce the level of adaptation. A recent study detected this effect using data on protein sequence evolution and codon usage in Drosophila. PMID- 12546810 TI - Apoptosis: mitochondrial membrane permeabilization--the (w)hole story? AB - One critical step of apoptosis is the release of mitochondrial proteins through the outer mitochondrial membrane. Recent work shows that two pro-apoptotic Bcl-2 family proteins, Bax and Bid, as well as the mitochondrion-specific lipid cardiolipin may cooperate in chemically defined liposomes to generate a protein permeable conduit, relaunching the debate on the identity of the pore responsible for mitochondrial membrane permeabilization during apoptosis. PMID- 12546811 TI - Region-specific apoptosis limits neural stem cell proliferation. AB - Regulation of stem cell division is of particular interest, both for studies of development and for stem cell therapeutics. In this issue of Neuron, Bello et al. show that the number of divisions of Drosophila neural stem cells is limited, in a region-specific manner, by regulated apoptosis in response to a pulse of expression of the Hox gene abdominal-A (abdA). PMID- 12546812 TI - Feeding hungry neurons: astrocytes deliver food for thought. AB - Among the proposed roles for astrocytes in the CNS is nutritive support for neurons. In this issue of Neuron, Voutsinos-Porche et al. provide evidence that astrocyte uptake of synaptic glutamate triggers astrocytic glycolysis and release of lactate, which in turn nourishes neurons and sustains neuronal activity. PMID- 12546813 TI - A computational basis to object? AB - To use an object, we must be able to perceive the spatial relationship between the object's parts. The accepted view of how the brain coherently encodes an object is that some neurons in the frontal cortex employ an object-centered coordinate frame. A new computational model challenges this view, using the rich conceptual framework of neural basis functions. PMID- 12546814 TI - Working smarter, not harder. AB - The ability to chunk, or to strategically organize, information is one of the most powerful ways people have to encode experience into memory. A functional brain imaging study by Bor and colleagues, in this issue of Neuron, provides evidence that prefrontal cortex contributes to this essential mnemonic process. PMID- 12546815 TI - Targeting dystroglycan in the brain. AB - The dystrophin-glycoprotein complex is a multisubunit complex that connects the extracellular matrix components to the cytoskeletal matrix of muscle fiber cells and is required for muscle integrity. Mutations in this complex are associated with muscular dystrophy. Although the role of dystroglycan has been explored mainly in the context of muscle, recent work has also demonstrated a novel role for dystroglycan in the CNS and thus provides potential insights into the brain abnormalities associated with some forms of muscular dystrophy. PMID- 12546816 TI - Autoregulation of neurogenesis by GDF11. AB - In the olfactory epithelium (OE), generation of new neurons by neuronal progenitors is inhibited by a signal from neurons themselves. Here we provide evidence that this feedback inhibitory signal is growth and differentiation factor 11 (GDF11). Both GDF11 and its receptors are expressed by OE neurons and progenitors, and GDF11 inhibits OE neurogenesis in vitro by inducing p27(Kip1) and reversible cell cycle arrest in progenitors. Mice lacking functional GDF11 have more progenitors and neurons in the OE, whereas mice lacking follistatin, a GDF11 antagonist, show dramatically decreased neurogenesis. This negative autoregulatory action of GDF11 is strikingly like that of its homolog, GDF8/myostatin, in skeletal muscle, suggesting that similar strategies establish and maintain proper cell number during neural and muscular development. PMID- 12546817 TI - A pulse of the Drosophila Hox protein Abdominal-A schedules the end of neural proliferation via neuroblast apoptosis. AB - Postembryonic neuroblasts are stem cell-like precursors that generate most neurons of the adult Drosophila central nervous system (CNS). Their capacity to divide is modulated along the anterior-posterior body axis, but the mechanism underlying this is unclear. We use clonal analysis of identified precursors in the abdomen to show that neuron production stops because the cell death program is activated in the neuroblast while it is still engaged in the cell cycle. A burst of expression of the Hox protein Abdominal-A (AbdA) specifies the time at which apoptosis occurs, thereby determining the final number of progeny that each neuroblast generates. These studies identify a mechanism linking the Hox axial patterning system to neural proliferation, and this involves temporal regulation of precursor cell death rather than the cell cycle. PMID- 12546818 TI - Axonal targeting of olfactory receptor neurons in Drosophila is controlled by Dscam. AB - Different classes of olfactory receptor neurons (ORNs) in Drosophila innervate distinct targets, or glomeruli, in the antennal lobe of the brain. Here we demonstrate that specific ORN classes require the cell surface protein Dscam (Down Syndrome Cell Adhesion Molecule) to synapse in the correct glomeruli. Dscam mutant ORNs frequently terminated in ectopic sites both within and outside the antennal lobe. The morphology of Dscam mutant axon terminals in either ectopic or cognate targets was abnormal. Target specificity for other ORNs was not altered in Dscam mutants, suggesting that different ORNs use different strategies to regulate wiring. Multiple forms of Dscam RNA were detected in the developing antenna, and Dscam protein was localized to developing ORN axons. We propose a role for Dscam protein diversity in regulating ORN target specificity. PMID- 12546819 TI - Pet-1 ETS gene plays a critical role in 5-HT neuron development and is required for normal anxiety-like and aggressive behavior. AB - The central serotonin (5-HT) neurotransmitter system is an important modulator of diverse physiological processes and behaviors; however, the transcriptional mechanisms controlling its development are largely unknown. The Pet-1 ETS factor is a precise marker of developing and adult 5-HT neurons and is expressed shortly before 5-HT appears in the hindbrain. Here we show that in mice lacking Pet-1, the majority of 5-HT neurons fail to differentiate. Remaining ones show deficient expression of genes required for 5-HT synthesis, uptake, and storage. Significantly, defective development of the 5-HT system is followed by heightened anxiety-like and aggressive behavior in adults. These findings indicate that Pet 1 is a critical determinant of 5-HT neuron identity and implicate a Pet-1 dependent program in serotonergic modulation of behavior. PMID- 12546820 TI - VRILLE feeds back to control circadian transcription of Clock in the Drosophila circadian oscillator. AB - The Drosophila circadian oscillator consists of interlocked period (per)/timeless (tim) and Clock (Clk) transcriptional/translational feedback loops. Within these feedback loops, CLK and CYCLE (CYC) activate per and tim transcription at the same time as they repress Clk transcription, thus controlling the opposite cycling phases of these transcripts. CLK-CYC directly bind E box elements to activate transcription, but the mechanism of CLK-CYC-dependent repression is not known. Here we show that a CLK-CYC-activated gene, vrille (vri), encodes a repressor of Clk transcription, thereby identifying vri as a key negative component of the Clk feedback loop in Drosophila's circadian oscillator. The blue light photoreceptor encoding cryptochrome (cry) gene is also a target for VRI repression, suggesting a broader role for VRI in the rhythmic repression of output genes that cycle in phase with Clk. PMID- 12546821 TI - Rapid induction of dendritic spine morphogenesis by trans-synaptic ephrinB-EphB receptor activation of the Rho-GEF kalirin. AB - The morphogenesis of dendritic spines, the major sites of excitatory synaptic transmission in the brain, is important in synaptic development and plasticity. We have identified an ephrinB-EphB receptor trans-synaptic signaling pathway which regulates the morphogenesis and maturation of dendritic spines in hippocampal neurons. Activation of the EphB receptor induces translocation of the Rho-GEF kalirin to synapses and activation of Rac1 and its effector PAK. Overexpression of dominant-negative EphB receptor, catalytically inactive kalirin, or dominant-negative Rac1, or inhibition of PAK eliminates ephrin induced spine development. This novel signal transduction pathway may be critical for the regulation of the actin cytoskeleton controlling spine morphogenesis during development and plasticity. PMID- 12546822 TI - Glial glutamate transporters mediate a functional metabolic crosstalk between neurons and astrocytes in the mouse developing cortex. AB - Neuron-glia interactions are essential for synaptic function, and glial glutamate (re)uptake plays a key role at glutamatergic synapses. In knockout mice, for either glial glutamate transporters, GLAST or GLT-1, a classical metabolic response to synaptic activation (i.e., enhancement of glucose utilization) is decreased at an early functional stage in the somatosensory barrel cortex following activation of whiskers. Investigation in vitro demonstrates that glial glutamate transport represents a critical step for triggering enhanced glucose utilization, but also lactate release from astrocytes through a mechanism involving changes in intracellular Na(+) concentration. These data suggest that a metabolic crosstalk takes place between neurons and astrocytes in the developing cortex, which would be regulated by synaptic activity and mediated by glial glutamate transporters. PMID- 12546823 TI - Extracellular Ca2+ depletion contributes to fast activity-dependent modulation of synaptic transmission in the brain. AB - Synaptic activation is associated with rapid changes in intracellular Ca(2+), while the extracellular Ca(2+) level is generally assumed to be constant. Here, using a novel optical method to measure changes in extracellular Ca(2+) at high spatial and temporal resolution, we find that brief trains of synaptic transmission in hippocampal area CA1 induce transient depletion of extracellular Ca(2+). We show that this depletion, which depends on postsynaptic NMDA receptor activation, decreases the Ca(2+) available to enter individual presynaptic boutons of CA3 pyramidal cells. This in turn reduces the probability of consecutive synaptic releases at CA3-CA1 synapses and therefore contributes to short-term paired-pulse depression of minimal responses. This activity-dependent depletion of extracellular Ca(2+) represents a novel form of fast retrograde synaptic signaling that can modulate glutamatergic information transfer in the brain. PMID- 12546824 TI - Excitatory actions of GABA in the cortex. AB - Little is known about how GABAergic inputs interact with excitatory inputs under conditions that maintain physiological concentrations of intracellular anions. Using extracellular and gramicidin perforated-patch recording, we show that somatic and dendritic GABA responses in mature cortical pyramidal neurons are depolarizing from rest and can facilitate action potential generation when combined with proximal excitatory input. Dendritic GABA responses were excitatory regardless of timing, whereas somatic GABA responses were inhibitory when coincident with excitatory input but excitatory at earlier times. These excitatory actions of GABA occur even though the GABA reversal potential is below action potential threshold and largely uniform across the somato-dendritic axis, and arise when GABAergic inputs are temporally or spatially isolated from concurrent excitation. Our findings demonstrate that under certain circumstances GABA will have an excitatory role in synaptic integration in the cortex. PMID- 12546825 TI - Mechanisms of gamma oscillations in the hippocampus of the behaving rat. AB - Gamma frequency oscillations (30-100 Hz) have been suggested to underlie various cognitive and motor functions. Here, we examine the generation of gamma oscillation currents in the hippocampus, using two-dimensional, 96-site silicon probes. Two gamma generators were identified, one in the dentate gyrus and another in the CA3-CA1 regions. The coupling strength between the two oscillators varied during both theta and nontheta states. Both pyramidal cells and interneurons were phase-locked to gamma waves. Anatomical connectivity, rather than physical distance, determined the coupling strength of the oscillating neurons. CA3 pyramidal neurons discharged CA3 and CA1 interneurons at latencies indicative of monosynaptic connections. Intrahippocampal gamma oscillation emerges in the CA3 recurrent system, which entrains the CA1 region via its interneurons. PMID- 12546826 TI - fMRI activation in response to illusory contours and salient regions in the human lateral occipital complex. AB - Regions in the human Lateral Occipital Complex (LOC) show fMRI responses to illusory surfaces. We show that the LOC activation is due to the globally completed region and occurs even when the region is not bounded by illusory contours (ICs). Kanizsa-type stimuli were modified by rounding the corners of the "pacmen" inducers and misaligning them slightly. The impression of an enclosed, salient region (SR) remained, although ICs were no longer perceived (psychophysical data). fMRI activity was elevated for both the IC and SR stimuli, compared to their control stimuli. The LOC response to salient regions may be the result of fast but crude region-based segmentation processes, which are useful for selecting parts of cluttered images for more detailed, computationally intensive processing. PMID- 12546827 TI - Integration of local features into global shapes: monkey and human FMRI studies. AB - The integration of local image features into global shapes was investigated in monkeys and humans using fMRI. An adaptation paradigm was used, in which stimulus selectivity was deduced by changes in the course of adaptation of a pattern of randomly oriented elements. Accordingly, we observed stronger activity when orientation changes in the adapting stimulus resulted in a collinear contour than a different random pattern. This selectivity to collinear contours was observed not only in higher visual areas that are implicated in shape processing, but also in early visual areas where selectivity depended on the receptive field size. These findings suggest that unified shape perception in both monkeys and humans involves multiple visual areas that may integrate local elements to global shapes at different spatial scales. PMID- 12546828 TI - Basis functions for object-centered representations. AB - In an object-centered representation, the position of the subparts of an object are encoded with respect to a set of axes and an origin centered on the object. Several physiological and neuropsychological results support the existence of such representations in humans and monkeys. An explicit representation would involve neurons with invariant response properties in object-centered coordinates. We consider an alternative scheme using basis functions in which the cells have retinotopic receptive fields modulated by the orientation of the object and task-related signals. We show that this alternative is consistent with single-cell data, is computationally efficient, and accounts for object-centered hemineglect, a syndrome observed in humans after fronto-parietal lesions. PMID- 12546829 TI - Encoding strategies dissociate prefrontal activity from working memory demand. AB - It is often proposed that prefrontal cortex is important in organization and control of working memory contents. In some cases, effective reorganization can decrease task difficulty, implying a dissociation between frontal activity and basic memory demand. In a spatial working memory task, we studied the improvement of performance that occurs when materials can be reorganized into higher level groups or chunks. Structured sequences, encouraging reorganization and chunking, were compared with unstructured sequences. Though structured sequences were easier to remember, event-related functional magnetic resonance imaging (fMRI) showed increased activation of lateral frontal cortex, in particular during memory encoding. The results show that, even when memory demand decreases, organization of working memory contents into higher level chunks is associated with increased prefrontal activity. PMID- 12546830 TI - Copycat viruses outwit the immune system. PMID- 12546832 TI - New rodent models gnawing at the black box of ALS. PMID- 12546831 TI - Do all paths lead to DARPP-32? PMID- 12546835 TI - Raising funding for early stage start-up companies. PMID- 12546836 TI - Hello Drug Discovery, I am from Insilico, take me to your President. PMID- 12546837 TI - Patent opportunities in bioinformatics. PMID- 12546838 TI - Blood hot in Boston. PMID- 12546840 TI - New ways to treat bacterial infections. AB - There is an urgent need for fresh approaches to the treatment of bacterial infections because of the changing patterns of infectious disease and the emergence of bacterial strains resistant to current antibiotics. Modification of the cell phenotype to sensitize bacteria to components of the hosts' immune system or to previously ineffective antibiotics could prevent the emergence of the resistant genotype. In addition, the use of light-activated antibacterial agents and lytic bacteriophage specific for key pathogens should be considered as safe and inexpensive alternatives to conventional treatment regimens for certain non-systemic infections. PMID- 12546841 TI - Rapid molecular theranostics in infectious diseases. AB - The increasing availability of rapid and sensitive nucleic acid testing assays for infectious diseases will revolutionize the practice of medicine by gradually reducing the need for standard culture-based microbiological methods that take at least two days. Molecular theranostics in infectious diseases is an emerging concept in which molecular biology tools are used to provide rapid and accurate diagnostic assays to enable better initial management of patients and more efficient use of antimicrobials. Essential conditions and the quality control required for the development and validation of such molecular theranostic assays are reviewed. PMID- 12546845 TI - Spying on nature's drug factories. PMID- 12546842 TI - Distribution and licensing of drug discovery tools--NIH perspectives. AB - Now, more than ever, drug discovery conducted at industrial or academic facilities requires rapid access to state-of-the-art research tools. Unreasonable restrictions or delays in the distribution or use of such tools can stifle new discoveries, thus limiting the development of future biomedical products. In grants and its own research programs the National Institutes of Health (NIH) is implementing its new policy to facilitate the exchanges of these tools for research discoveries and product development. PMID- 12546846 TI - Military surveillance system for breast cancer detection. PMID- 12546847 TI - Inhalant induces tolerance against stroke. PMID- 12546850 TI - Rat of the month. PMID- 12546851 TI - Formulation of therapeutic synthetic polymers for drug and gene delivery. PMID- 12546852 TI - Mining the human "kinome". PMID- 12546853 TI - Genetic approach to chemical genetics. PMID- 12546855 TI - Raising venture capital in the biopharma industry. AB - Raising venture capital (VC) is both an art and a science. Future entrepreneurs should carefully consider the various issues of VC financing that have a strong impact on the success of their business. In addition to attracting the best venture capital firms, these issues include such subtle but important points as the timing of financing (especially of the first round), external support sources, desirable qualities of a VC firm, amount to be raised, establishing a productive interface between the founders and the venture capitalists, and most importantly the effects of well-executed VC funding on hiring senior executives and scientific leaders. PMID- 12546856 TI - Chromosomal biomarkers of genomic instability relevant to cancer. AB - It is generally acknowledged that a crucial event in the initiation and evolution of cancer is the acquisition of a genomic instability phenotype. This review focuses on mechanisms of chromosomal instability including aneuploidy, chromosome rearrangement and breakage-fusion-bridge cycles. The role of micronutrient deficiency, such as folate deficiency, in the causation of chromosomal instability is briefly reviewed and the concept of recommended dietary allowances for genomic stability is introduced. In addition, the techniques for measuring the various chromosomal instability events are discussed with a focus on the cytokinesis-block micronucleus assay as an almost complete system for measuring these various genetic mishaps. PMID- 12546857 TI - Drug targeting to specific vascular sites. AB - The blood vessels of individual tissues are biochemically distinct, and pathological lesions put their own signature on the vasculature. In tumors, both blood and lymphatic vessels differ from normal vessels. New methods, such as in vivo screening of phage libraries, have provided peptides and antibodies that recognize these vascular signatures and can be used in targeted delivery of therapeutic agents. Targeting a therapy to the diseased tissue enhances the efficacy of the treatment while reducing the side effects in mouse experiments. Results from drug delivery to tumor vessels have been particularly encouraging. PMID- 12546859 TI - Catalytic antioxidants prevent type 1 diabetes. PMID- 12546861 TI - Prion vaccine: is there hope against CJD? PMID- 12546860 TI - Energy blocker to treat liver cancer. PMID- 12546864 TI - Should scientific innovation be managed? PMID- 12546865 TI - Predicting novel proteins and their interactions. PMID- 12546866 TI - From screen-saver to virtual screener: harnessing latent PC power through distributed computing. PMID- 12546867 TI - Systems biology: the new darling of drug discovery? PMID- 12546869 TI - Surrogate endpoints in cancer drug development. AB - Over the next three years a large number of novel, mechanistically targeted drugs will enter clinical trials for cancer. The remarkable progress in understanding the molecular biology of cancer has provided an enormous range of validated targets for drug discovery. Following lead optimisation and suitable pharmaceutical formulation these compounds have undergone rapid screening in preclinical models. Innovative methods of clinical development are now essential to ensure optimal dose determination and scheduling. The discovery of novel surrogates for efficacy is essential in this fast moving area and requires imaginative partnerships between academic groups and the pharmaceutical industry. PMID- 12546870 TI - Discovery informatics: its evolving role in drug discovery. AB - Drug discovery and development is a highly complex process requiring the generation of very large amounts of data and information. Currently this is a largely unmet informatics challenge. The current approaches to building information and knowledge from large amounts of data has been addressed in cases where the types of data are largely homogeneous or at the very least well defined. However, we are on the verge of an exciting new era of drug discovery informatics in which methods and approaches dealing with creating knowledge from information and information from data are undergoing a paradigm shift. The needs of this industry are clear: Large amounts of data are generated using a variety of innovative technologies and the limiting step is accessing, searching and integrating this data. Moreover, the tendency is to move crucial development decisions earlier in the discovery process. It is crucial to address these issues with all of the data at hand, not only from current projects but also from previous attempts at drug development. What is the future of drug discovery informatics? Inevitably, the integration of heterogeneous, distributed data are required. Mining and integration of domain specific information such as chemical and genomic data will continue to develop. Management and searching of textual, graphical and undefined data that are currently difficult, will become an integral part of data searching and an essential component of building information- and knowledge-bases. PMID- 12546871 TI - Permeability issues in nasal drug delivery. AB - The nasal route is one of the most permeable and highly vascularized site for drug administration ensuring rapid absorption and onset of therapeutic action. It has been potentially explored as an alternative route for drugs with poor bioavailability and for the delivery of biosensitive and high molecular weight (MW) compounds such as proteins, peptides, steroids, vaccines, and so on. This review discusses the major factors affecting the permeability of drugs or biomolecules through the nasal mucosa, including biological, formulation and device-related factors. This information could potentially help to achieve desired plasma concentrations of drugs without compromising or altering the normal physiology of the nasal cavity. PMID- 12546874 TI - Expanding the HTS paradigm. PMID- 12546875 TI - In need of high-throughput behavioral systems. AB - One of the current major bottlenecks in drug discovery is in vivo testing of candidate drugs in behavioral paradigms in normal or genetically altered mice. This testing is essential in discovering gene function and predicting potential efficacy of CNS drugs in humans. New efforts in the biotech community aim to alleviate this bottleneck by developing higher-throughput systems of behavioral, neurological and physiological analyses. Together with large pharmacological databases, equipped with state-of-the-art bioinformatic and/or data-mining algorithms, these systems will provide rapid and accurate indices of the therapeutic potential of novel drugs. By providing a substantial increase in the speed of behavioral testing, new high-throughput systems will facilitate current behavioral research with faster, more reliable approaches. Furthermore, screening whole drug-libraries and comparing the profiles of novel compounds to those of known compounds will facilitate the discovery of novel drugs. Target validation will also become more efficient with the fast characterization of novel mutant mice. PMID- 12546876 TI - Applications of transfected cell microarrays in high-throughput drug discovery. AB - DNA microarrays and, more recently, protein microarrays, have become important tools for high-throughput genomic and proteomic studies. Transfected cell microarrays are a complementary technique in which array features comprise clusters of cells overexpressing defined cDNAs. Complementary DNAs cloned in expression vectors are printed on microscope slides, which become living arrays after the addition of a lipid transfection reagent and adherent mammalian cells. This article discusses two potential uses of cell microarrays in drug discovery: as a method of screening for gene products involved in biological processes of pharmaceutical interest and as in situ protein microarrays for the development and assessment of leads. PMID- 12546877 TI - The impact of protein biochips and microarrays on the drug development process. AB - With the genome sequences of several organisms now in public databases, the scientific community has realized that it is time to prepare for the next step: the understanding of biological systems or systems biology. Whereas genes contain the information for life, the encoded proteins and RNAs fulfill nearly all the functions, from replication to regulation. At present, there is a perceived demand for high-throughput and parallel analytical devices as research tools in systems biology, and, in addition, for new concepts to extract knowledge and value from these data. Protein biochips will play a decisive role in meeting this need in the future. PMID- 12546878 TI - High-throughput cell-based assays in yeast. AB - With the sequencing of many genomes now completed, biologists are faced with the challenge of deciphering the function and association of an immense number of predicted proteins. Comprehension of the networks of proteins and chemicals that comprise the cells and tissues of an organism, and the specific roles of proteins in these networks, will be a necessary next step to understanding cellular function in healthy and diseased states. In the past decade, the budding yeast Saccharomyces cerevisiae has emerged as an important tool for large-scale functional genomics analyses. This review describes the use of yeast cell-based assays in the post-genomic era, focusing on high-throughput functional genomics and drug discovery. PMID- 12546879 TI - High-throughput cell analysis using multiplexed array technologies. AB - The desire for more biologically relevant data from primary screening has resulted in a dramatic increase of cell-based assays in HTS labs. Consequently, new cell-array technologies are being developed to increase the quality and quantity of data emerging from such screens. These technologies take the form of both positional and non-positional formats, each with their own advantages. Notably, screens using these technologies generate databases of high-quality data that can be analyzed in ways currently not possible. The power of cell-based assays combined with new array and analytical technologies will enable the condensation of serial drug discovery processes, thereby decreasing the time and cost of taking a hit compound into clinical trials. Here, we compare array strategies being developed towards the goal of integrating multiplexed cell-based assays into HTS. PMID- 12546880 TI - Emerging high-throughput drug target validation technologies. AB - Identifying the right target for drug development is a critical bottleneck in the pharmaceutical and biotech industries. The genomics revolution has shifted the problem from a scarcity of targets to a surplus of putative drug targets. As the validity of a target cannot be simply inferred from correlative data, the key is confirmation of the causative role of a gene product in a particular disease. It should therefore be recognized that an effective therapeutic strategy requires an appropriate target validation technology to verify the right target. PMID- 12546881 TI - Antibody arrays: an embryonic but rapidly growing technology. AB - Protein arrays are now an attractive proposition as they can measure a diverse range of protein interactions not possible with traditional DNA arrays. Antibody arrays are a specific subset of this technology. Originally conceived as multi analyte detectors, antibody arrays are now used in a wide variety of applications. For instance, the potential of this technology to diagnose human diseases, such as leukemia, breast cancer and, potentially, heart failure, has stimulated much interest. Furthermore, identification of new protein targets in particular disease states will prove to be an invaluable tool in drug discovery and development. Patient prognosis and treatment are also potential applications of the technology. Antibody arrays have proved to be dynamic in response to these broad range of possibilities. This review examines variations in antibody array design and discusses current and potential applications of this novel and interesting technology. PMID- 12546882 TI - Novel detection strategies for drug discovery. AB - The Human Genome Project is expected to increase the number of potential drug targets from the current figure of 500 to approximately 3,000-4,000. This increased number of targets, and increasing knowledge of signaling-pathway networks and their complexities, sets new demands for efficiency on HTS assay technologies. Assessment of the total efficacy of a given drug candidate requires not only the classical assays, but also a wide variety of assays related to signaling cascades and cellular functions. Discrete functional assays traditionally involved Ca(2+) flux, kinases and cAMP, but today extend to the whole signaling network, from ligand binding to expression. This review discusses emerging novel non-radioisotopic assays, such as ligand-stimulated GTP-binding, the inositol triphosphate assay, cellular receptor trafficking, and protein protein interactions. PMID- 12546883 TI - Pretzel-shaped peptide for autoimmune disease. PMID- 12546884 TI - Boost for retinoic acid cancer therapy. PMID- 12546885 TI - Top of the form. PMID- 12546888 TI - Microbes, viruses and verse--microbial musings. PMID- 12546889 TI - Towards a reconciliation between molecular diagnostics and new pharmaceuticals. PMID- 12546890 TI - Affinity fingerprints--leading the way? PMID- 12546891 TI - Anti-infectives in the 21st century. PMID- 12546893 TI - RNAi: gene-silencing in therapeutic intervention. AB - Several rapidly developing RNA interference (RNAi) methodologies hold the promise to selectively inhibit gene expression in mammals. RNAi is an innate cellular process activated when a double-stranded RNA (dsRNA) molecule of greater than 19 duplex nucleotides enters the cell, causing the degradation of not only the invading dsRNA molecule, but also single-stranded (ssRNAs) RNAs of identical sequences, including endogenous mRNAs. As such, RNAi technology is currently being evaluated not only as an extremely powerful instrument for functional genomic analyses, but also as a potentially useful method to develop highly specific dsRNA based gene-silencing therapeutics. PMID- 12546894 TI - Structure-based virtual screening: an overview. AB - Enormous advances in genomics have resulted in a large increase in the number of potential therapeutic targets that are available for investigation. This growth in potential targets has increased the demand for reliable target validation, as well as technologies that can identify rapidly several quality lead candidates. Virtual screening, and in particular receptor-based virtual screening, has emerged as a reliable, inexpensive method for identifying leads. Although still an evolving method, advances in computational techniques have enabled virtual screening to have a positive impact on the discovery process. Here, the current strengths and weaknesses of the technology are discussed, and emphasis is placed on aspects of the work-flow of a virtual screening campaign, from preparation through to post-screening analysis. PMID- 12546895 TI - Application of hydrogen bonding calculations in property based drug design. AB - A drug can be characterized by "descriptors" that include size (volume) and H bond acidity and H-bond basicity. These descriptors can be rapidly estimated from structure by a fragment scheme and used to predict physicochemical and transport properties of drug candidates (e.g. logP, solubility, gastrointestinal absorption, permeability and blood-brain distribution). The solvation equations can be interpreted to provide a qualitative chemical insight into biological partition and transport mechanisms. Applications to blood-brain partition and human intestinal absorption (HIA) are discussed. PMID- 12546898 TI - Industrialized molecular biology, information biotechnology, and the blockbuster drug model--alive and well at age 50. PMID- 12546899 TI - Knowledge assembly for the life sciences. PMID- 12546900 TI - Rational drug discovery: what can we learn from regulatory networks? AB - To enable the list of genes and proteins contained within genomic databases to be useful for drug discovery, we need to understand how the genome maps into the phenome. An essential, but not explicitly listed ingredient of the genome is the regulatory interactions between genes and proteins that form a genome-wide network. How can the concept of regulatory networks increase our understanding of living systems? Networks are more than just static "wiring diagrams". Gene interactions impose dynamic constraints, which, although obvious in emergent phenotypic properties, are not captured by traditional one gene-one trait approaches. Understanding the nature of these constraints in gene-activation state space will pave the way to a holistic yet formal and genomics-based approach to rational drug development. PMID- 12546901 TI - From microarrays to networks: mining expression time series. AB - Over the past few years, powerful new methods have been devised that enable researchers to study the expression dynamics of many genes simultaneously (e.g. gene expression profiles using cDNA microarrays). In principle, this potentially vast quantity of data enables the dissection of the complex genetic networks that control the patterns and rhythms of gene expression in the cell. Finding the patterns in those data represents the next major phase in our understanding of the programming and functioning of the living cell. Simple dynamic models can be used to generate gene expression networks. These networks reveal the phenomenological link between the expression of different genes. This review discuss how these networks are generated and outlines several data-mining techniques for extracting relationships and hypotheses in gene expression. These emerging methods can be applied to a range of biological problems. PMID- 12546902 TI - Grid technologies empowering drug discovery. AB - Grid technologies enable flexible coupling and sharing of computers, instruments and storage. Grids can provide technical solutions to the volume of data and computational demands associated with drug discovery by delivering larger computing capability (flexible resource sharing), providing coordinated access to large data resources and enabling novel online exploration (coupling computing, data and instruments online). Here, we illustrate this potential by describing two applications: the use of desktop PC grid technologies for virtual screening, and distributed X-ray structure reconstruction and online visualization. PMID- 12546903 TI - Leveraging process integration in early drug discovery. AB - Recent advances in new analysis and prediction concepts in informatics, statistics and computational chemistry have drawn attention to mining the enormous flood of information generated from ultra-high-throughput screening (uHTS) and early drug discovery more effectively. This review analyses current infrastructure and process concepts in data analysis, storage and mining, with a particular focus on high-throughput technologies. It also provides examples of how these techniques have been applied successfully together with underlying reasons for these developments. PMID- 12546904 TI - Effective experimental design: enzyme kinetics in the bioinformatics era. AB - Acquiring details about the kinetic parameters of enzymes is crucial to both drug development and clinical diagnosis. The correct design of an experiment is crucial to collecting data suitable for analysis, modelling and deriving the correct information. As classical design methods are not targeted to the more complex kinetics now frequently studied, further work is required to estimate parameters of such models with low variance. This review examines the different options available to produce major gains in information, productivity and the accuracy of each experiment. PMID- 12546905 TI - Small- and large-scale biosimulation applied to drug discovery and development. AB - Biosimulation uses mathematics to quantitatively represent the dynamics of biological systems and thereby analyze and predict system behavior. Biosimulations can be classified into two general categories: small-scale models designed to address a specific problem, and large-scale models of detailed regulatory mechanisms used to address a broad scope of questions. Both classes of biosimulations have been applied to problems important for drug discovery and development. Small-scale biosimulations have been particularly useful for interpreting clinical data and developing novel biomarkers. Large-scale biosimulations typically integrate a wide variety of data and can provide insights into how complex biological systems are regulated in both health and disease. Because large-scale biosimulations represent detailed regulatory mechanisms and their interactions, they can predict the overall clinical effect of modulating individual pathways or targets. In this mini-review, we describe several examples of how small- and large-scale biosimulations have been applied to problems important for drug development in diabetes, HIV, heart disease and asthma. PMID- 12546906 TI - Biomedical informatics: the future for drug development. AB - The problems that exist in drug development are well documented: the limited number of new chemical entities, increased cost of drug development, problems in clinical trials (Phase III), product launches that result in withdrawal, and pressure to reduce the cost of pharmaceuticals from the government. It appears that the promise of genomics has not yet reached its full potential to impact the process. This review identifies the need to develop and implement the area of biomedical informatics for increased success in drug development and healthcare in general. PMID- 12546907 TI - Of mice and men, and cancer research. PMID- 12546909 TI - Enlightening cholera bug reveals new virulence system. PMID- 12546908 TI - Muscular dystrophy: toxic RNA to blame. PMID- 12546912 TI - Rolf Krebs discusses the pharma industry past, present and future. Interview by Joanne Clough. PMID- 12546913 TI - Promises of text processing: natural language processing meets AI. PMID- 12546914 TI - The semantic web and biology. PMID- 12546915 TI - Immunotherapies: cause for measured optimism. PMID- 12546917 TI - Therapeutic synthetic polymers: a game of Russian roulette? AB - Synthetic polymer-based drug-delivery systems have been applied in drug delivery for the past 50 years. So why are there so few examples of these macromolecules being used successfully in the clinic? It is our view that many products are failing because of a neglect of the fundamental science surrounding the architectural control of the molecules present, their behaviour following in vivo administration and host response. Adverse events following parenteral administration of approved synthetic polymer-based systems have resulted in unpredictable and fatal responses in a significant number of individuals. Acceptance of the importance of immunotoxicological factors in response to the presence of these macromolecules must be addressed if emergent technologies, such as polymer-based gene-delivery systems, are going to succeed. PMID- 12546918 TI - High-throughput methods for the development of new catalytic asymmetric reactions. AB - Chiral, single enantiomer pharmaceuticals have become increasingly more important. Therefore, research aimed at providing new methods for their selective preparation has taken on an even greater importance. One of the most efficient strategies for the synthesis of non-racemic, chiral molecules is asymmetric catalysis. There are many variables involved in the discovery of a new catalytic asymmetric transformation; hence, methods for the rapid screening of large numbers of catalysts have been developed. Herein, these techniques and strategies for the rapid discovery of novel asymmetric catalysts are reviewed. PMID- 12546919 TI - Selecting protein tyrosine phosphatases as drug targets. AB - Protein tyrosine phosphatases (PTPs) have emerged as a new and promising class of signaling targets, since the discovery of PTP1B as a major drug target for diabetes and obesity. Blocking individual PTPs results in the activation of specific tyrosine phosphorylation events, but matching PTPs with such pathways and therapeutic indications is a complex undertaking. The history of PTP1B shows that its unusual knockout phenotype and observations with generic and antisense inhibitors in vivo, but not its classical molecular biology, triggered the rapid development of inhibitors that are today being developed for the clinic. PMID- 12546922 TI - Killer snails ease the pain. PMID- 12546923 TI - Edible vaccines against human papilloma virus. PMID- 12546924 TI - Regulatable gene therapy. PMID- 12546928 TI - Mnemonics--the art of recollection. PMID- 12546930 TI - Gaps in text-based knowledge discovery for biology. PMID- 12546929 TI - Mistletoe lectins: telomerase inhibitors in alternative cancer therapy. PMID- 12546931 TI - Cheminformatics--decision making in drug discovery. PMID- 12546932 TI - New routes for drug discovery. PMID- 12546933 TI - Why do we need so many chemical similarity search methods? AB - Computational tools to search chemical structure databases are essential to finding leads early in a drug discovery project. Similarity methods are among the most diverse and most useful. We will present some lessons we have gathered over many years experience with in-house methods on several therapeutic problems. The effectiveness of any similarity method can vary greatly from one biological activity to another in a way that is difficult to predict. Also, any two methods tend to select different subsets of actives from a database, so it is advisable to use several search methods where possible. PMID- 12546934 TI - Applications of antisense and siRNAs during preclinical drug development. AB - A significantly greater number of candidate drug targets and compounds are now being generated during preclinical drug development. To date, however, such increases have not led to improvements in clinical success rates or reduced times to market. There is a need for better strategies to prioritize targets and drug candidates. Antisense and siRNA technologies offer exceptional speed and specificity to address this need. In particular, antisense and siRNAs are beginning to be used in combination with expression profiling to evaluate drug specificity and mechanism-of-action, aiding in the identification of better candidates earlier in the drug development process. PMID- 12546939 TI - Spin-outs and incubators: philosophy and practice. PMID- 12546935 TI - Substrate SARs in human P450s. AB - Drug metabolism is now an integral part of the drug discovery process, and the cytochromes P450 (CYPs) are the most important family of enzymes involved in human drug metabolism. An increased understanding of the properties of the substrates for the major human CYPs is thus highly desirable. This article shows how key characteristics of CYP substrates, such as lipophilicity, molecular mass and hydrogen-bonding potential, govern selectivity towards individual CYPs. Importantly, the variation in binding affinities of 60 human CYP substrates can be explained by understanding the key physicochemical, structural and electronic characteristics that govern substrate binding to each isozyme. PMID- 12546940 TI - Antisense inhibitor provides new treatment approach for hypercholesterolaemia. PMID- 12546941 TI - Gene therapy progress for HIV. PMID- 12546943 TI - Potential new vaccine against chlamydia. PMID- 12546947 TI - The dilemma of process development. PMID- 12546948 TI - Live vectors: are safe but effective vaccines possible? PMID- 12546949 TI - Want novel drugs in a hurry? You had better do the math! PMID- 12546950 TI - A new era as plate movers get on track. PMID- 12546952 TI - Integrating diagnostics and therapeutics: revolutionizing drug discovery and patient care. AB - Over the next five years it is widely anticipated that the molecular diagnostics industry will continue to grow at double-digit pace to meet increasing demand for personalized medicine. A wide variety of drugs in late preclinical and early clinical development is now being targeted to disease-specific gene and protein defects that will require co-approval of diagnostic and therapeutic products by regulatory agencies. For clinical laboratories and pathologists, this integration of diagnostics and therapeutics represents a major new opportunity to emerge as leaders of the new medicine, guiding the selection, dosage, route of administration and multi-drug combinations, and producing increased efficacy and reduced toxicity of pharmaceutical products. PMID- 12546953 TI - Function first: a powerful approach to post-genomic drug discovery. AB - In the post-genomic era, pharmaceutical researchers must evaluate vast numbers of protein sequences and formulate novel, intelligent strategies for identifying valid targets and discovering leads against them. The identification of small molecules that selectively target proteins or protein families will be aided by knowing the function and/or the structure of the target(s). By identifying protein function first, efficiencies are gained that allow subsequent focus of resources on particular protein families of interest. This article reviews current proteomic-scale approaches to identifying function as a way of accelerating lead discovery. PMID- 12546954 TI - Novel chemical genetic approaches to the discovery of signal transduction inhibitors. AB - Concurrent advances in both high-throughput chemistry and genomics have given rise to the field of chemical genetics as a discipline for elucidating and validating drug targets, and generating novel therapeutics. Indeed, chemical genetic approaches to drug discovery have now been applied to several important drug target classes, especially those involved in signal transduction. Chemical genetics is distinct from the broader term "chemogenomics" which is defined as the description of all possible drugs against all possible targets (reviewed in [1]). This review covers several "orthogonal" chemical genetic approaches and focuses on a unique analog sensitive kinase technology and its applications to kinase drug discovery. PMID- 12546957 TI - Economics, pharmacoeconomics and drug discovery for dementias. PMID- 12546958 TI - Gene therapy trials for cystic fibrosis. PMID- 12546959 TI - CDK inhibitor shows promise for inflammatory kidney disease. PMID- 12546963 TI - Samir Hanash discusses how HUPO aims to globalize proteomics research (interview by Joanna Owens). AB - Samir Hanash is a pioneer in cancer proteomics and was elected as the Inaugural President of the Human Proteome Organization (HUPO) in June 2001. He is Professor of Medicine at the University of Michigan (UM) where he heads a large research group that focuses on cancer proteomics, and is also on the executive committees of several research associations, including the American Society for Cell Biology (ASCB) and the American Association for Cancer Research (AACR). After obtaining a medical degree in 1972 from American University in Beirut, Lebanon, Hanash moved to UM where he studied for his PhD on haemoglobin protein and gene analysis in the Human Genetics department and was awarded the Public Health Service (PHS) Fellowship award in Biochemical Genetics in 1975. After a post-doctoral fellowship in the same department, Hanash took up a Residency position in Pediatrics at the Children's Hospital of Michigan, during which time he was awarded a Children's Leukaemia Foundation Fellowship Award. Subsequently, he has held the positions of Instructor in Pediatric Hematology (1978-1979), and Assistant Professor (1979-1984) and Associate Professor (1984-1989) of Pediatrics before taking on his present role as Professor of Pediatrics at UM. He is also Director of the Cancer Center Carcinogenesis Program and a member of the Gene Therapy Group and the Genome Center and Computer Task Force at UM. He was awarded the Rothchild Award by the Curie Cancer Institute in 1998. [interview by Joanna Owens] PMID- 12546960 TI - Pharmacogenomics suggests new treatment approach for leukaemia. PMID- 12546964 TI - Protein kinase drugs--optimism doesn't wait on facts. PMID- 12546965 TI - De novo chemical ligand design. PMID- 12546966 TI - Macroresults through microarrays. PMID- 12546968 TI - Chemoproteomics as a basis for post-genomic drug discovery. AB - The large number of small organic compounds now available for drug-lead screening has led to numerous methods for classifying molecular similarity and diversity, the aim being to restore a balance between the quantity and drug-like quality of compounds in small-molecule libraries. Whereas structural and physicochemical attributes continue to be emphasized in compound selection for drug-lead screening, chemoproteomics--the use of biological information to guide chemistry- offers a highly efficient alternative to small-molecule characterization that can accelerate drug discovery in the post-genomic era. PMID- 12546969 TI - Protein microarray technology. AB - Microarray technology allows the simultaneous analysis of thousands of parameters within a single experiment. Microspots of capture molecules are immobilised in rows and columns onto a solid support and exposed to samples containing the corresponding binding molecules. Readout systems based on fluorescence, chemiluminescence, mass spectrometry, radioactivity or electrochemistry can be used to detect complex formation within each microspot. Such miniaturised and parallelised binding assays can be highly sensitive, and the extraordinary power of the method is exemplified by array-based gene expression analysis. In these systems, arrays containing immobilised DNA probes are exposed to complementary targets and the degree of hybridisation is measured. Recent developments in the field of protein microarrays show applications for enzyme-substrate, DNA-protein and different types of protein-protein interactions. This article discusses theoretical advantages and limitations of any miniaturised capture-molecule ligand assay system and discusses how the use of protein microarrays will change diagnostic methods and genome and proteome research. PMID- 12546970 TI - Modified release drug delivery in veterinary medicine. AB - To successfully research and develop an animal pharmaceutical dosage form, a diverse array of issues covering basic medicine, pharmacology and technology must be addressed. Societal concerns regarding animal and public health, as well as the rapidly changing farming and economic environments, provide additional challenges that require integration into an already complex web of issues. Here, we examine the drive towards reducing the frequency of administration to animals and the closing of gaps between the human and veterinary drug product development. PMID- 12546975 TI - Solving the problem of antimicrobial resistance: is a global approach necessary? PMID- 12546976 TI - Cervical dysplasia: encapsulated DNA treatment shows promise. PMID- 12546977 TI - New immunotherapy triple strategy for AML. PMID- 12546978 TI - Targeting cell migration. PMID- 12546981 TI - Chris Lipinski discusses life and chemistry after the Rule of Five. PMID- 12546982 TI - Current understanding of mistletoe lectins. PMID- 12546983 TI - Polymers for drug delivery: more research needed. PMID- 12546985 TI - Pharmaceutical companies need to broaden research. PMID- 12546986 TI - GPCRs come of age in San Diego. AB - Highlights from IBC Life Science's 7th annual G-protein coupled receptors, 14-16 October 2002 in San Diego, CA, USA. PMID- 12546988 TI - Examining unmet needs in infectious disease. AB - In the past 30 years, more than 30 new aetiological agents of infectious disease have been identified. Some of these are responsible for entirely novel and life threatening disorders, such as AIDS, Ebola fever, hantavirus pulmonary syndrome and Nipah virus encephalitis. During the same period, some longstanding infectious diseases (such as tuberculosis) have became resurgent, as a result of a combination of complacency, increased travel and social dislocation, and also increasing drug resistance. This review looks at some of the key unmet needs in this therapeutic area and discusses strategies to address them. PMID- 12546989 TI - The identification of clinically relevant markers and therapeutic targets. AB - The history of tumor biomarker discovery has been one of limited success. Population based screens are few and of limited clinical usefulness. Biomarkers that are able to segregate patients by diagnosis, prognosis and appropriate therapeutic selection are in great need and will be the basis of the clinical management in the future. This review sets out the challenges inherent in the field of tumor biomarker discovery and the tools that we are using to meet that challenge. It is now possible, using this suite of technologies, to discuss novel tumor biomarkers in terms of a pipeline rather than single unique events in research. The future of clinical oncology management will use markers such as those being identified via these techniques to improve patient care through better diagnosis and hopefully to achieve greater success in treatment by exploiting tumor markers as therapeutic targets. PMID- 12546990 TI - Gene discovery and validation for neurodegenerative diseases. AB - Treatment of neurodegenerative diseases, such as Alzheimer's disease, multiple sclerosis, Parkinson's disease and amyotrophic lateral sclerosis (ALS), represents a major challenge for the pharmaceutical industry. These disorders have common and unique molecular pathological characteristics that result in serious reductions in nervous-system functionality. Key to developing novel and efficacious therapeutics is the discovery of new gene targets. Genomic, proteomics and bioinformatic analyses are identifying vast amounts of genes whose expression is associated with the pathology of a specific disease. Extensive validation studies performed in parallel with drug development are crucial for the selection of appropriate target genes. This review outlines some of the current progress in gene discovery for neurodegenerative disease. PMID- 12546995 TI - Neural stem cells as novel drug delivery agents. PMID- 12546994 TI - Benzodiazepine library yields lupus target. PMID- 12546996 TI - AIDS therapy: more harm than help? PMID- 12546999 TI - Fortifying the over forties. AB - Raymond C. Rowe, in his private prescription column, waxes lyrical on the subject of fortification for the over forties. PMID- 12547000 TI - Polymers--a synthetic or natural choice? PMID- 12547001 TI - Polymer therapeutics--what is missing? PMID- 12547003 TI - Targeting signal transduction with large combinatorial collections. AB - The large-scale application of combinatorial chemistry to drug discovery is an endeavor that is now more than ten years old. The growth of chemical libraries together with the influx of novel genomic targets has led to a reconstruction of the drug-screening paradigm. The drug discovery industry faces a post-genomic world where the interplay between tens-of-thousands of proteins must be addressed. To compound this complexity, there now exists the ability to screen millions of compounds against a single target. This review focuses on the practice and use of selecting individual compounds from large chemical libraries that act on targets relevant to signal transduction. PMID- 12547004 TI - Structural pharmacogenomics, drug resistance and the design of anti-infective super-drugs. AB - Large-scale comparative analysis of drug-target polymorphism structures enables the rational design of next generation 'super drugs'--drugs that are less prone to development of drug resistance or that work for the largest possible fraction of the patient population. Furthermore, knowledge of the drug-target-shape repertoire that exists within the patient population enables predictions of likely clinical trial outcomes and response rates for drug efficacy. This gives information on the optimal drug candidates before the initiation of clinical trials. The economic impact of incorporating pharmacogenomics insights early on in the drug discovery process will be substantial and will afford significant competitive advantages to companies that successfully incorporate this technology. PMID- 12547005 TI - Aptamers as tools for target prioritization and lead identification. AB - The increasing number of potential drug target candidates has driven the development of novel technologies designed to identify functionally important targets and enhance the subsequent lead discovery process. Highly specific synthetic nucleic acid ligands--also known as aptamers--offer a new exciting route in the drug discovery process by linking target validation directly with HTS. Recently, aptamers have proven to be valuable tools for modulating the function of endogenous cellular proteins in their natural environment. A set of technologies has been developed to use these sophisticated ligands for the validation of potential drug targets in disease models. Moreover, aptamers that are specific antagonists of protein function can act as substitute interaction partners in HTS assays to facilitate the identification of small-molecule lead compounds. PMID- 12547008 TI - Biofilm studies yield targets against cystic fibrosis. PMID- 12547009 TI - Chemical geneticists unify their data. PMID- 12547010 TI - Cold war adversaries team up to conquer TB. PMID- 12547013 TI - Conducting research ethically in developing countries. PMID- 12547014 TI - Parenteral polymers. PMID- 12547015 TI - Diagnostics meets therapeutics: the impact of pharmacogenetics. PMID- 12547016 TI - Meeting the challenges in screening. PMID- 12547017 TI - Selective retinoids and rexinoids in cancer therapy and chemoprevention. AB - Natural and synthetic retinoids are effective inhibitors of tumor cell growth in vitro and in vivo. However, the toxicity of natural derivatives of vitamin A limits their therapeutic use. Recently, synthetic compounds selective for the different retinoid receptor isotypes have been generated that circumvent pan retinoid toxicity. The tumor-suppressive activity of selective retinoid and/or rexinoid ligands has been established preclinically, and emerging clinical trials are supportive of the chemotherapeutic and chemopreventive potential of these compounds in multiple oncology indications, with reduced toxicity. Moreover, the combination of retinoids and/or rexinoids with chemotherapeutic agents for the synergistic modulation of specific pathways could also be of benefit in cancer therapy. PMID- 12547018 TI - Random mutagenesis in the mouse as a tool in drug discovery. AB - The flood of raw information generated by large-scale data acquisition technologies in genomics, microarrays and proteomics is changing the early stages of the drug discovery process. Although many more potential drug targets are now available compared with the pre-genomics era, knowledge about the physiological context in which these targets act--information crucial to both discovery and development--is scarce. Random mutagenesis strategies in the mouse provide scalable approaches for both the gene-driven validation of candidate targets in vivo and the discovery of new physiological pathways by phenotype-driven screens. PMID- 12547019 TI - Nasal drug delivery: new developments and strategies. AB - The use of the nasal route for the delivery of challenging drugs has created much interest in recent years in the pharmaceutical industry. Consequently, drug delivery companies are actively pursuing the development of novel nasal drug delivery systems and the exploitation of these for administration of conventional generic drugs and peptides, both in-house and with partners in the pharmaceutical industry. This review sets out to discuss some new developments and strategies in nasal drug delivery. An exiting discovery that drugs can be transported directly from nose to brain via the olfactory pathway is discussed and examples of proof of-concept in man are given. PMID- 12547022 TI - Holographic imaging of live tumours. PMID- 12547021 TI - Molecular imaging: what picture does it paint for future oncology? PMID- 12547023 TI - New cannabinoid for multiple sclerosis. PMID- 12547024 TI - Dumped drugs could prevent HIV. PMID- 12547027 TI - A personalised limerick--evidence of esteem? PMID- 12547028 TI - The double-edged cytokine sword of non-viral gene targeting to tumors. PMID- 12547029 TI - The importance of predictive ADME simulation. PMID- 12547030 TI - Aggressive outsourcing yields therapeutic breadth. PMID- 12547032 TI - High-throughput protein expression for the post-genomic era. AB - In the past, protein expression has been perceived as the principle bottleneck in protein characterization and structure determination. The challenge now is to rapidly express large numbers of genes in the search for new drug targets and therapeutic proteins encoded by the human genome. In this competitive environment, several high-throughput expression strategies for protein production are being used to industrialize the process of protein expression. PMID- 12547033 TI - Barriers to remember: brain-targeting chemical delivery systems and Alzheimer's disease. AB - Brain-targeted chemical delivery systems (CDSs) represent rational drug design attempts not only to deliver but also to target drugs to their site of action. Using a sequential metabolism approach, the special bidirectional properties of the blood-brain barrier can be exploited to smuggle the precursors of therapeutic compounds across the barrier and lock them inside the brain ready for sustained release of the active drugs. Many potential therapeutic applications can be envisioned for such CDSs; here, the potential of brain-targeted estradiol for the prevention and treatment of Alzheimer's disease is reviewed in detail. PMID- 12547034 TI - Sterols and sterolins: new drugs for the immune system? AB - Since the discovery of glucocorticoids, we have had a single strategy for manipulating the immune system in cases of destructive diseases mediated by uncontrolled immune responses. However, long-term use of immunosuppressive drugs can lead to the threat of opportunistic infections and malignancies. As we learn more about regulatory subsets of T lymphocytes and their cytokine profiles, the thrust has been on developing new ligands that ultimately give us more site specific control. Our group has developed a patented mixture of plant sterols and sterolins that has anti-inflammatory properties and profound immune modulating effects on subsets of CD4+ T cells. We have tested this mixture in several clinical entities and we believe that it has wide applications in reverting immune abnormalities. PMID- 12547037 TI - Diseases with chronic stage in a population with varying size. AB - An epidemiological model of hepatitis C with a chronic infectious stage and variable population size is introduced. A non-structured baseline ODE model which supports exponential solutions is discussed. The normalized version where the unknown functions are the proportions of the susceptible, infected, and chronic individuals in the total population is analyzed. It is shown that sustained oscillations are not possible and the endemic proportions either approach the disease-free or an endemic equilibrium. The expanded model incorporates the chronic age of the individuals. Partial analysis of this age-structured model is carried out. The global asymptotic stability of the infection-free state is established as well as local asymptotic stability of the endemic non-uniform steady state distribution under some additional conditions. A numerical method for the chronic-age-structured model is introduced. It is shown that this numerical scheme is consistent and convergent of first order. Simulations based on the numerical method suggest that in the structured case the endemic equilibrium may be unstable and sustained oscillations are possible. Closer look at the reproduction number reveals that treatment strategies directed towards speeding up the transition from acute to chronic stage in effect contribute to the eradication of the disease. PMID- 12547038 TI - A mathematical model for the investigation of the Th1 immune response to Chlamydia trachomatis. AB - Chlamydia are bacterial pathogens of humans and animals causing the important human diseases trachoma, sexually transmitted chlamydial disease and pneumonia. Of the human chlamydial diseases, sexually transmitted disease caused by Chlamydia trachomatis is a major public health concern. Chlamydia trachomatis replicates intracellularly and is characterised by a complex developmental cycle. Chlamydia is susceptible to humoral and cell-mediated immunity. Here we investigate the Th1 cell-mediated immune response against Chlamydia-infected cells as the response changes over the chlamydial developmental cycle. We suggest a form for the immune response over one developmental cycle by modelling the change in the number of intracellular chlamydial particles and assume peptides are presented in proportion to the number of replicating forms of chlamydial particles. We predict, perhaps non-intuitively, that persistent Chlamydia should be induced and forced not to return to the lytic cycle. We also suggest that extending the length of the time of the lytic cycle will effectively decrease the required efficacy of the Th1 response to eliminate the pathogen. We produce plots of active disease progression, control and clearance for varying levels of Th1 effectiveness. PMID- 12547039 TI - Simulation of waves in calcium models with 3D spherical geometry. AB - Waves of calcium ions are present in fertilized eggs of many species. Models for pulse and tidal wave propagation have usually been studied in one or two spatial coordinates only. We examine in three spatial coordinates some established models, based on Ca(2+)-induced Ca(2+)-release from both (assumed) continuously or heterogeneously distributed stores of endoplasmic reticulum (ER) through channels activated by inositol triphosphate (IP(3)). With continuous IP(3) distribution decreasing radially towards the interior, we obtain concave pulse shapes for waves penetrating the interior. Concave waves are also recorded in systems with ER confined to distributions of small spheres (microdomains) inside the cell, which we simulate for front waves (tides) in bistable systems. PMID- 12547040 TI - A new approach of fitting biomass dynamics models to data. AB - A non-traditional approach of fitting dynamic resource biomass models to data is developed in this paper. A variational adjoint technique is used for dynamic parameter estimation. In the variational formulation, a cost function measuring the distance between the model solution and the observations is minimized. The data assimilation method provides a novel and computationally efficient procedure for combining all available information, i.e., the data and the model in the analysis of a resource system. This technique will be used to analyze data for the North-east Arctic cod stock. Two alternative population growth models: the logistic and the Gompertz model are used for estimating parameters of simple bioeconomic models by the method of constrained least squares. Estimates of the parameters of the models dynamics are reasonable and can be accepted. The main inference from the work is that the average fishing mortality is found to be significantly above the maximum sustainable yield value. PMID- 12547041 TI - Design principles and operating principles: the yin and yang of optimal functioning. AB - Metabolic engineering has as a goal the improvement of yield of desired products from microorganisms and cell lines. This goal has traditionally been approached with experimental biotechnological methods, but it is becoming increasingly popular to precede the experimental phase by a mathematical modeling step that allows objective pre-screening of possible improvement strategies. The models are either linear and represent the stoichiometry and flux distribution in pathways or they are non-linear and account for the full kinetic behavior of the pathway, which is often significantly effected by regulatory signals. Linear flux analysis is simpler and requires less input information than a full kinetic analysis, and the question arises whether the consideration of non-linearities is really necessary for devising optimal strategies for yield improvements. The article analyzes this question with a generic, representative pathway. It shows that flux split ratios, which are the key criterion for linear flux analysis, are essentially sufficient for unregulated, but not for regulated branch points. The interrelationships between regulatory design on one hand and optimal patterns of operation on the other suggest the investigation of operating principles that complement design principles, like a user's manual complements the hardwiring of electronic equipment. PMID- 12547042 TI - Epidemiological modeling in a branching population. Particular case of a general SIS model with two age classes. AB - This paper covers the elaboration of a general class of multitype branching processes for modeling in a branching population, the evolution of a disease with horizontal and vertical transmissions. When the size of the population may tend to infinity, normalization must be carried out. As the initial size tends to infinity, the normalized model converges a.s. to a dynamical system the solution of which is the probability law of the state of health for an individual ancestors line. The focal point of this study concerns the transient and asymptotical behaviors of a SIS model with two age classes in a branching population. We will compare the asymptotical probability of extinction on the scale of a finite population and on the scale of an individual in an infinite population: when the rates of transmission are small compared to the rate of renewing the population of susceptibles, the two models lead to a.s. extinction, giving consistent results, which no longer applies to the opposite situation of important transmissions. In that case the size of the population plays a crucial role in the spreading of the disease. PMID- 12547043 TI - Re: CT-guided neurolytic splanchnic nerve block. PMID- 12547044 TI - Efficacy of gelclair in reducing pain in palliative care patients with oral lesions: preliminary findings from an open pilot study. PMID- 12547045 TI - Brain natriuretic peptide: a marker for dyspnea? PMID- 12547046 TI - Understanding the continuum of palliative care for patients and their caregivers. AB - We describe a process for assisting seriously ill patients and their caregivers in prioritizing their preferences for care during advanced illness. Thirty-two seriously ill patients and their caregivers participated in seven 90-minute focus groups conducted cross-sectionally in Denver, Colorado; San Francisco, California; and Washington State. Fourteen expert-defined end-of-life quality indicators were presented to each group, and quantitative unweighted rankings were obtained through patient and caregiver preferences. Aggregated weightings were used to rank the top five quality measures for exploration of open-ended questions. Pain management was the most important quality indicator among all three groups. Overlap in preferences were found for at least two of the groups for symptom management, monitoring medical issues, advance care preferences and assessment of family and caregiver involvement. Caregivers were more focused on bereavement support. Variation in preferences for end-of-life care by patients and their caregivers reflects the need to understand the individual decision making regarding end-of-life care. Quality improvement activities are beginning to address these needs. PMID- 12547047 TI - Sleep and quality of life in breast cancer patients. AB - This study described sleep in a heterogeneous sample of breast cancer patients using the Pittsburgh Sleep Quality Index (PSQI) and examined the relation between sleep disturbance and health-related quality of life as measured by the Rand 36 Item Health Survey. Chemotherapy and radiation therapy were explored as predictors of sleep disturbance in breast cancer patients, and the sleep characteristics of breast cancer patients were compared to the sleep characteristics of a sample of medical patients with general medical conditions. Results showed that 61% of breast cancer patients had significant sleep problems. Sleep was characterized by reduced total sleep time with sleep frequently being disturbed by pain, nocturia, feeling too hot, and coughing or snoring loudly. Despite the frequency of significant sleep disturbance, pharmacological and cognitive-behavioral treatments of sleep problems were observed to be inadequate. Limited evidence was found for the role of chemotherapy and radiation therapy in the sleep disturbance of breast cancer patients, and the general pattern of sleep disturbance in breast cancer patients was not significantly different than that observed in medical patients with general medical conditions. Breast cancer patients having significant sleep problems had greater deficits in many areas of health-related quality of life. The implications of the findings and study limitations are discussed. PMID- 12547048 TI - Patient-related barriers to fatigue communication: initial validation of the fatigue management barriers questionnaire. AB - Fatigue is a highly prevalent and distressing symptom of cancer and its treatment. However, cancer patients often fail to communicate with their oncologists about fatigue. In this study, we attempted to identify the patient related barriers to communication about fatigue, as cited by patients. Two hundred patients were sampled across the Community Cancer Care, Inc. (CCC) network of Indiana using the Cancer Behavior Inventory-Brief scale (CBI-B), the Zung Self-Rating Depression Scale (ZSDS), the Functional Assessment of Cancer Inventory-Fatigue scale (FACT-F), and the Fatigue Management Barriers Questionnaire (FMBQ), a questionnaire devised by experts in the field of cancer related fatigue. There were no significant correlations between the instrument scores and demographic variables. Scores on the instruments did not differ significantly based on whether the patient was from a rural or urban site. One hundred thirty-two patients (66%) reported that they had never spoken to their doctor about fatigue. The most frequently reported reasons for this lack of patient communication about fatigue included the doctor's failure to offer interventions (47%), patients' lack of awareness of effective treatments for fatigue (43%), a desire on the patient's part to treat fatigue without medications (40%), and not wanting to complain to the doctor (28%). Patients reported that medical staff offered a mean of 11.63 recommendations for dealing with fatigue. The FMBQ was found to correlate significantly with self-efficacy (CBI-B, r = -0.20, P < 0.01) and correlate weakly with the number of recommendations made (r = -0.15, P < 0.05). The FMBQ was noted to have acceptable internal consistency (alpha = 0.88) and validity and may prove to be a useful instrument for understanding why patients do not communicate about fatigue. Multiple barriers contribute to why cancer patients do not comment about fatigue but may be overcome if physicians screen and assess for this symptom. PMID- 12547049 TI - Longitudinal documentation of cancer pain management outcomes: a pilot study at a VA medical center. AB - We measured pain outcomes in a cohort of patients with cancer pain in a general hematology/oncology setting at a Veterans Administration Medical center (VA). The outcomes included pain relief, pain severity, changes in pain severity, interference scores, symptom distress, quality of life (QOL), and satisfaction. Seventy-four (74) consecutive patients with worst cancer-related pain equal to or greater than 4/10 were recruited. Cancer pain diagnoses were made and the cancer pain management guidelines of the United States Agency for Health Care Policy and Research were followed. Patients were followed weekly using the Brief Pain Inventory (BPI), medication diary, satisfaction questionnaire, visual analogue quality of life scale (VASQOL) and record of side effects for 3 weeks. The Functional Assessment of Cancer Therapy (FACT-G) and Memorial Symptom Assessment Scale Short Form (MSAS-SF) were used at initial and final interviews. The mean initial worst pain severity was 8.3 (range 4-10) and mean pain relief was 40% (range 0-100). By week 1, the majority of patients achieved pain relief of >/=80%, with a corresponding decrease in worst pain severity and pain interference scores. Pain continued to decrease over three weeks. At week 3, there was a significant improvement in the MSAS-SF psychological symptom distress subscale (P = 0.02). The average number of opioid-related side effects was 5 and remained steady over time. Most patients felt "quite a bit" or "very much" satisfied at all weeks. There was a significant improvement in VASQOL (P < 0.005) and in FACTG SUMQOL scores (P = 0.007). This experience demonstrates that cancer pain management can result in measurable and significant changes in pain relief, pain severity, pain interference scores, psychological symptom distress, and QOL scores. PMID- 12547050 TI - Obtaining informed consent for cancer pain research: do patients with advanced cancer and patients with chronic pain have different concerns? AB - To explore the factors that patients with malignant and nonmalignant pain consider when deciding whether to enroll in pain research studies, determine whether their views are different, and determine whether willingness to enroll in research is associated with pain severity, semi-structured interviews were conducted with 80 patients (cancer pain: n = 40; chronic nonmalignant pain: n = 40). The risks and potential benefits that were important to patients with cancer were the same as those that were important to patients with chronic pain. Willingness to enroll in research was associated with pain severity (Spearman rho = 0.33; P = 0.041) in patients with chronic pain, but not in patients with cancer pain. Patients with cancer pain do not have different concerns than chronic pain patients do. Although chronic pain patients' willingness to enroll in research was related to pain severity and a desire for better pain management, cancer patients' willingness to enroll was not. PMID- 12547051 TI - The Norwegian brief pain inventory questionnaire: translation and validation in cancer pain patients. AB - The European Association of Palliative Care recommends the Brief Pain Inventory questionnaire (BPI) as a pain assessment tool in clinical studies. After translation into Norwegian, we administered the BPI to 300 hospitalized cancer patients. Cronbach's alphas were computed to assess reliability, and factor analysis was utilized to ascertain construct validity. The BPI interference and pain severity scales were validated against items on pain intensity and pain influence on daily function in the European Organization for Research and Therapy of Cancer (EORTC) QLQ-C30 questionnaire. In total, 235 patients (78%) were able to complete the BPI questionnaire, but 82 (35%) of these questionnaires had one or more missing items. Cronbach's alphas were 0.87 for the pain severity and 0.92 for the interference scales. A factor analysis identified three factors; pain intensity, interference with physical function, and interference with psychological functions/sleep. These three factors explained 82% of the variance. The correlation between BPI pain severity index and the EORTC QLQ-C30 item on pain intensity was 0.70 (P < 0.001). The correlation between BPI interference index and the EORTC QLQ-C30 item on pain influence on daily living was 0.62 (P < 0.001). We conclude that BPI has satisfactory psychometric properties, but is not completed by a significant proportion of patients. Further research is needed to establish pain assessment tools for patients unable to answer a comprehensive pain questionnaire, to establish routines for analysis of missing values, and to investigate if pain interference items also reflect disease-related impairment. PMID- 12547052 TI - Cancer pain and psychosocial factors: a critical review of the literature. AB - Poor pain assessment is cited as one barrier to the adequate treatment of cancer pain. The identification of relevant psychosocial factors may improve the assessment of chronic cancer pain. This article presents: 1) a critical review of the evidence for an association between chronic cancer pain and psychological distress, social support, and coping; 2) clinical implications of the findings; and 3) recommendations for future research. Fourteen of the 19 reviewed studies on psychological distress found a significant association between increased pain and increased distress. Seven of the eight studies on social support found significant association between higher levels of pain and decreased levels of social activities and social support. Three of the four studies that examined coping strategies found that increased catastrophizing was significantly associated with more intense pain. Based on several criteria, the evidence is considered Strong for psychological distress, Moderate for social support, and Inconclusive for coping. This review suggests that comprehensive chronic pain assessment should include routine screening for psychological distress. PMID- 12547053 TI - Lidocaine toxicity during frequent viscous lidocaine use for painful tongue ulcer. AB - Oral viscous lidocaine is useful for the treatment of symptoms induced by oral inflamed mucosa, such as radiation- or chemotherapy-induced mucositis. The toxic reactions associated with an accidental overdose have been reported in pediatric cases. We report a case of lidocaine toxicity in a 22-year-old man during frequent viscous lidocaine use for severe painful tongue ulcer. The toxic symptoms developed when the amount of oral viscous lidocaine exceeded 240 ml per day. The serum lidocaine concentration associated with this use was 6.7 microg/ml. The toxic symptoms continued in spite of the serum lidocaine concentration below the toxic level after the start of a diluted preparation, which contained a half-dose lidocaine. It is speculated that lidocaine metabolites might have contributed to the toxic symptoms. Clinicians should consider the risk of lidocaine toxicity in cases of frequent viscous lidocaine use, and determine the serum concentrations of lidocaine and its metabolites. PMID- 12547149 TI - Allelic imbalance in selected chromosomal regions in ovarian cancer. AB - Ovarian cancer (OC) is often asymptomatic at the initial stage. When diagnosed, up to 75% of the patients present grade III or IV tumors with metastasis in nearby organs of the abdomen. Genetic imbalance is abundant in OC, and allelic loss (AL) of specific chromosomal regions is considered an early event. To establish association between genetic markers for early diagnosis/prognosis of OC, our target was to define narrow specific regions of AL. We analyzed 65 ovarian carcinomas by using 19 microsatellite markers located in three different chromosomes. First, a 7.6-Mb region containing the estrogen receptor (ESR1) and the tumor suppressor gene LATS1 was analyzed. Several chromosomal breakpoints flanking ESR1 affecting the region harboring LATS1 were found. Second, we found chromosomal breakpoints on 13q13.1 approximately q13.3 that defined two narrow regions flanking the BRCA2 locus. Third, our ovarian tumors exhibited a very high frequency of AL on 16q and chromosomal breakpoints defining two narrow regions within 16q22.2 approximately q24.3. In this article, we report three new polymorphic microsatellite markers and strong evidence of AL of narrow well defined regions in hot spots on 6q, 13q, and 16q in ovarian tumors. PMID- 12547150 TI - Hematologic malignancies and Klinefelter syndrome. a chance association? AB - Klinefelter syndrome was first described in 1942 as an endocrine disorder characterized by gynecomastia, hypogonadism, small testes, and elevated levels of follicle-stimulating hormone. An extra X chromosome (i.e., 47,XXY) was subsequently demonstrated in these patients and an increased incidence of leukemia and lymphoma has been described. We report a retrospective study of a series of unselected patients with Klinefelter syndrome diagnosed by cytogenetic studies and the occurrence of hematologic malignancies. The literature is also reviewed. PMID- 12547151 TI - Late appearance of t(1;19)(q11;q11) in myelodysplastic syndrome associated with dysplastic eosinophilia and pulmonary alveolar proteinosis. AB - We report a case of myelodysplastic syndrome (MDS), which developed marked eosinophilia and pulmonary alveolar proteinosis after the appearance of t(1;19)(q11;q11). Chromosomal analysis of the peripheral eosinophils identified the same chromosome abnormality in all metaphases to that of bone marrow blast cells. Review of the literature revealed three reported cases of concurrent MDS and pulmonary alveolar proteinosis. We reviewed four cases of concurrent MDS and pulmonary alveolar proteinosis, including the present case. Interestingly, all but one of these patients also had evidence of eosinophilia and abnormality of chromosome 1p. These findings, together with morphologic abnormalities of eosinophils observed in this case, indicate clonal involvement of eosinophils in the MDS clone, and that the eosinophilia was derived from the neoplastic clone with the translocation. We postulate that this chromosomal rearrangement is involved in the development of eosinophilia and pulmonary alveolar proteinosis in MDS. PMID- 12547152 TI - Genomic imbalances in ovarian borderline serous and mucinous tumors. AB - We analyzed 25 ovarian borderline tumors (13 serous and 12 mucinous tumors) by comparative genomic hybridization (CGH). Genomic imbalance was detected in 85% of serous tumors and 75% of mucinous tumors. Different patterns of genomic alterations were identified in serous and mucinous tumors. Gain of the X chromosome was common in both serous (30%) and mucinous (42%) tumors. However, gain of chromosome 8 was detected exclusively in 38% of serous and mixed sero mucinous tumors, but not in any pure mucinous tumors. According to the present and previous studies, gain of chromosome 8 is the most common abnormality in borderline serous tumors. Gain of the same chromosome is also common in high grade and advanced stage serous carcinomas, but uncommon in early stage serous carcinomas. In addition gain of chromosome X is common in borderline serous and mucinous tumors, while loss of chromosome X is predominant in invasive carcinomas. These findings do not support the multi-step progression theory from borderline tumor to high-grade, advanced stage carcinoma, but indicate that the borderline ovarian tumor is a distinct entity. Genes in chromosome 8 may be critical for the development and the differentiation of borderline serous tumors. PMID- 12547153 TI - A subgroup of malignant fibrous histiocytomas is associated with genetic changes similar to those of well-differentiated liposarcomas. AB - Increasing clinical and pathological evidence suggests that malignant fibrous histiocytomas (MFH) comprise a heterogeneous tumor group. In a series of 108 MFH tested by comparative genomic hybridization, we found in 22 tumors high-level coamplification of the 12q14 approximately q15 chromosome region with other loci, a genetics strongly reminiscent of what has been observed for well-differentiated liposarcomas. Nevertheless, these MFH differ from liposarcomas by a high recurrence of coamplified partners because coamplified loci were seen at 1p32 in nine cases, 6q23 in seven cases, and 12q24 in six cases. The same recurrence was observed in a series of dedifferentiated liposarcomas, but not in a series of well-differentiated liposarcomas. These observations demonstrate that a subgroup of MFH share a genetic partner very similar to that observed in liposarcomas, and suggest that the undifferentiated status of these tumors is closely related to the amplifications of specific chromosome loci. PMID- 12547154 TI - BCR/ABL amplification in chronic myelocytic leukemia blast crisis following imatinib mesylate administration. AB - The onset of accelerated phase or blast crisis of chronic myelocytic leukemia (CML) is usually associated with the acquisition of new chromosome abnormalities in addition to the t(9;22)(q34;q11) that is characteristic of the chronic phase CML. We describe the cytogenetic and molecular genetic findings in two cases of myelocytic blast crisis of CML, one occurring 6 months after commencing treatment with the ABL-specific tyrosine kinase inhibitor imatinib mesylate (STI571, Glivec, or Gleevec) and the second treated with imatinib mesylate for established blast crisis. In both cases, multiple secondary cytogenetic abnormalities were observed at transformation, with homogeneously staining regions that were shown to contain BCR/ABL amplification by fluorescence in situ hybridization appearing after imatinib mesylate administration. BCR/ABL amplification is emerging as an important mechanism of acquired resistance to imatinib mesylate. PMID- 12547155 TI - Novel clonal der(8)t(8;14)(p11;q11),del(9)(q13q22) and t(14;22) (q13;q13) in a patient with fulminant adult T-cell leukemia/lymphoma. AB - We describe a 70-year-old man with fulminant adult T-cell leukemia/lymphoma. He died of progressive disease 1 week after the onset of symptoms. The integrated viral DNA of human T-lymphotropic virus type I was detected in the bone marrow aspirate by polymerase chain reaction. Cytogenetic analysis of bone marrow cells showed novel clonal aberrations consisting of 46,XY,der(8)t(8;14)(p11;q11),del(9) (q13q22),t(14;22)(q13;q13). PMID- 12547156 TI - Clonal development of a blastoid mantle cell lymphoma studied with comparative genomic hybridization. AB - A molecular cytogenetic study was performed on the diagnostic tumor sample and three relapses from a case with blastoid mantle cell lymphoma. The clonal relatedness of the tumors was demonstrated by identical rearrangements of the immunoglobulin heavy chain gene and was supported by results from comparative genomic hybridization analyses. All samples shared the common alterations of losses of 6q, 9p, and 11q and gains of 3q, 9q, 12p, and 13q, suggesting that they were relatively early events in the tumorigenesis. Relapse 1 also showed a loss of 8p, while relapses 2 and 3 had gained the X chromosome and 7p, in addition, relapse 3 displayed gains of chromosomes 3 and 20. Taken together, the findings suggest that relapses 2 and 3 developed from the diagnostic tumor sample, while relapse 1 represents a separate lineage of tumor progression originating directly from a postulated ancestral tumor cell carrying the common chromosomal alterations identified in all tumors. PMID- 12547157 TI - Coamplification of 12p11 and 12q13 approximately q22 in multiple ring chromosomes in a spindle cell sarcoma resolved by novel multicolor fluorescence in situ hybridization analysis. AB - An 80-year-old male presented with a lobulated mass in the lower abdominal wall. A diagnosis of an intermediate grade myofibroblastic spindle cell sarcoma was made. Cytogenetic analysis demonstrated a complex karyotype with a der(6), a small marker and five, different in size, ring chromosomes. Fluorescence in situ hybridization (FISH), multiplex FISH, and multicolor banding analysis was used to further delineate this complex karyotype. The der(6) was shown to be a der(18)t(6;18;9;12;18), the marker chromosome was identified as del(17), and the ring chromosomes as r(9) and r(12;18)x4. Amplification of 18 and coamplification of 12p and 12q was detected in the ring and marker chromosomes. No intercellular heterogeneity was observed although a few micronuclei containing chromosome 18 and anaphase bridges, containing chromosome 12 material, the result of bridge fusion-bridge (BFB) cycles, were observed. Our findings combined with results from others indicate that amplification of chromosomes 12 and 18 as well as BFB phenomena characterize this type of sarcoma. PMID- 12547158 TI - Peripheral-type benzodiazepine receptor (PBR) gene amplification in MDA-MB-231 aggressive breast cancer cells. AB - Recent studies using human breast cancer cell lines, animal models, and human tissue biopsies have suggested a close correlation between the expression of the peripheral-type benzodiazepine receptor (PBR) and the progression of breast cancer. This study investigates the genetic status of the PBR gene in two human breast cancer cell lines: MDA-MB-231 cells, which are an aggressive breast cancer cell line that contains high levels of PBR, and MCF-7 cells, which are a nonaggressive cell line that contains low levels of PBR. Both DNA (Southern) blot and fluorescence in situ hybridization analyses indicate that the PBR gene is amplified in MDA-MB-231 relative to MCF-7 cells. These data suggest that PBR gene amplification may be an important indicator of breast cancer progression. PMID- 12547159 TI - Two balanced and novel chromosomal translocations in myeloid malignancies. characterization by multiplex fluorescence in situ hybridization. AB - We describe two novel chromosomal translocations in two cases of leukemia in which these translocations were further characterized as the sole acquired karyotypic abnormality by mutliplex fluorescence in situ hybridization (M-FISH). They comprised a case of acute myeloid leukemia with t(6;10)(q21;p12) and a case of chronic myelomonocytic leukemia with t(5;12)(q34;q24). To the best of our knowledge, these two balanced translocations are novel and are hitherto unrecognized in hematologic malignancies. While the clinical and pathogenic significance of these translocations remains to be defined, the present report illustrates that M-FISH technology contributes to the exclusion of subtle or cryptic translocations in sole karyotypic aberrations and the confirmation of novel chromosomal arrangements in neoplastic disorders. PMID- 12547160 TI - Is t(10;11)(p11.2;q23) involving MLL and ABI-1 genes associated with congenital acute monocytic leukemia? AB - Congenital, or perinatal, leukemias are rarely observed, but retrospective molecular studies seem to suggest a more frequent onset in prenatal life. Myelocytic types are common, and chromosome band 11q23 rearrangements at the MLL locus are characteristic genetic markers. The fusion of the MLL gene with one of its partners, ABI-1, has recently been described in two infant leukemia patients with monocytic involvement and good clinical outcome. We report a case of congenital monocytic leukemia with the same gene involvement and good response to chemotherapy. The blast metaphases were probed by fluorescence in situ hybridization, and t(10;11)(p11.2;q23) involving MLL and ABI-1 genes was demonstrated with the same breakpoint in ABI-1. The congenital presentation of this case suggests a possible relationship of this genetic event with in utero leukemogenesis. PMID- 12547161 TI - Partial deletion of chromosome 1 in a case of acute myelocytic leukemia. AB - Acute myelocytic leukemia (AML) is a malignant disease characterized by the proliferation of immature myelocytic precursor cells causing the disruption of normal bone marrow function. Many chromosomal aberrations have been described in AML including translocations, inversions, deletions, and additions. Here we describe a novel deletion of chromosome 1, del(1)(p34p36) in a case of AML, French-American-British classification M1, in a previously healthy 33-year-old male. This isolated cytogenetic abnormality occurred in 33% of the myeloblasts examined at diagnosis. Subsequent cytogenetic analyses conducted on marrow following induction and consolidation therapy demonstrated a normal male karyotype in all cells examined. The patient remains in clinical and hematological remission 22 months following diagnosis. The presence of 1p abnormalities in AML and other malignancies is reviewed, as are candidate tumor suppressor genes in the 1p34 approximately p36 region. The implications of chromosome 1p abnormalities on clinical outcome are also discussed. PMID- 12547162 TI - Novel dic(16;18)(q11;p11) in two cases of Philadelphia chromosome positive acute B-cell lymphoblastic leukemia. AB - We report two adult patients with Philadelphia chromosome positive (Ph+) B-cell acute lymphoblastic leukemia (ALL) who presented an additional dic(16;18)(q11;p11) that, to the best of our knowledge, has never been previously reported. Fluorescence in situ hybridization analysis confirmed the translocation and showed it to be dicentric. Both patients were treated for the ALL, but showed refractory disease and died despite aggressive treatment. Similarly to what has been reported with other additional chromosome abnormalities, our cases suggest that the presence of this novel translocation confers an adverse effect to the already poor prognosis of Ph+ ALL. PMID- 12547163 TI - Malignant peripheral primitive neuroectodermal tumor of the kidney. AB - Ewing family of tumors is a group of highly aggressive neoplasias that occur most commonly in the first two decades of life. These tumors are most frequently localized in bones, less frequently in soft tissues. They usually appear as undifferentiated small round-cell tumors. With current treatment regiments, 5 year disease-free survival rates exceed 60% in patients with a localized disease. Patients with metastatic disease at the time of their first presentation have a poor prognosis. We describe a rare case of visceral primitive neuroectodermal tumor with the involvement of the kidney in a 9-year-old girl. The tumor was studied with immunohistochemistry, cytogenetics, and molecular biology methods. Strong expression of protein MIC(2) by immunochemistry (antibody HBA 71) with subsequent demonstration of a translocation consistent with t(11;22)(q24;q12) using cytogenetic and reverse transcriptase polymerase chain reaction (RT-PCR) confirmed the histopathological diagnosis of peripheral primitive neuroectodermal tumor. We detected minimal residual disease in bone marrow using RT-PCR. PMID- 12547164 TI - Deletion 6q as a recurrent chromosomal aberration in T-cell large granular lymphocyte leukemia. AB - T-cell large granular lymphocyte (T-LGL) leukemia is an uncommon disease, with limited information on karyotypic aberrations. No consistent chromosomal changes have thus far been described. We report two cases of T-LGL leukemia who presented with severe anemia. The LGL were CD3+, CD4-, CD8+, CD56-, and CD161-, with variable expression of CD94, CD158a, and CD158b, and had clonal T-cell receptor gene rearrangement. A deletion of the long arm of chromosome 6 was the sole aberration in both cases. This is the first report of a recurrent chromosomal aberration in T-LGL leukemia. PMID- 12547165 TI - Constitutional rearrangements of 7q22 in hematologic malignancies. a new case report. AB - Abnormalities of chromosome 7 are a frequent finding in myelocytic malignancies and are associated with a poor prognosis. Based on chromosome banding analysis, two critical regions have been identified: one in band 7q22 and the second in region 7q32 approximately q35. The chromosomal breakpoint in band 7q22 appears to be heterogeneous and may involve tumor suppressor gene(s). Constitutional rearrangements of 7q22 have rarely been reported in myeloid malignancies. To our knowledge, this is the first report in the literature of a myeloproliferative disorder with a constitutional t(1;7)(q42;q22). PMID- 12547166 TI - Altered splicing pattern of TACC1 mRNA in gastric cancer. AB - Transforming acidic coiled-coil (TACC) proteins are centrosome and microtubule associated proteins that are essential for mitotic spindle function. We identified TACC1 as an immunogenic protein and a potential tumor antigen by applying serological identification of antigens by recombinant expression cloning (SEREX) technique to screen a gastric cancer cDNA library. The 5'RLM-RACE and reverse transcriptase polymerase chain reaction analyses revealed at least six different transcript variants of TACC1 with variable transcription start sites and alternative exon usage (designated TACC1-A-TACC1-F). All transcripts differ in their 5' ends but share an identical 3' region encoding coiled-coil domain. Four transcripts were universally expressed in all normal tissues analyzed but TACC1-D and TACC1-F showed a restricted expression pattern. TACC1-F, a transcript representing the SEREX-identified cDNA clone, was predominantly expressed in brain and gastric tumors to a similar level. TACC1-D was only weakly detectable in kidney and colon but not in other normal tissues, while a relatively strong expression was observed in 50% of gastric cancer tissue samples analyzed. These transcript variants are generated possibly as a result of alterations in efficiency and pattern of alternative splicing; these isoforms may represent genetic markers, for example TACC1-D for gastric cancer. We also propose that inappropriate expression of the isoforms in gastric cancer cells might result in dysfunction of TACC1 thus contributing to the genetic instability. PMID- 12547167 TI - Structural chromosomal abnormality of 1q in acute leukemia with Sweet syndrome. PMID- 12547168 TI - Identical karyotypes in synchronous bilateral clear cell renal cell carcinomas. PMID- 12547169 TI - Chromosome 9 interstitial deletion in multiple myeloma. PMID- 12547170 TI - Serum antibodies to Dicrocoelium dendriticum in sheep from Sardinia (Italy). AB - An indirect enzyme-linked immunosorbent assay (ELISA) with Dicrocoelium dendriticum excretory/secretory antigens was used to evaluate the presence of serum antibodies against the trematode in 738 sheep randomly chosen in Sardinia (Italy). Coprological sedimentation was used to discover egg-output. Seropositivity was detected in 86.2% tested sheep, whereas faecal prevalence was 6.7%; all that were faecal-positive also were ELISA-positive. PMID- 12547171 TI - A herd-level analysis of risk factors for antibodies to Sarcocystis neurona in Michigan equids. AB - Equine protozoal myeloencephalitis (EPM) is a neurological disease of horses and ponies caused by infection of the central nervous system with the protozoan parasite Sarcocystis neurona. A herd-level analysis of a cross-sectional study of serum antibodies to S. neurona in Michigan equids was conducted, using data collected in 1997 for study that included 1121 equids from 98 Michigan horse farms. Our objective was to identify specific herd-level risk factors associated with seropositivity. We tested associations between herd seroprevalence and various farm-management practices (including feed-storage methods and wildlife control). Multivariable models were developed for three strata based on relative opossum abundance (opossum districts). Herd seroprevalence ranged from 0 to 100% (median=57%). No risk factor was significantly associated with herd seroprevalence at P< or = 0.05 in all opossum districts. Our results suggest that equids living in areas with large opossum populations might be infected with S. neurona from multiple sources. PMID- 12547172 TI - Comparison of two atmospheric-dispersion models to assess farm-site exposure to sour-gas processing-plant emissions. AB - We describe two approaches for exposure assessment that we used in a large-scale retrospective cattle study conducted in Alberta, Canada. Sulfur dioxide (SO(2)) was the surrogate measure of exposure to a complex mixture of combusted sour-gas emissions. Monthly air pollution dispersion modeling (1985-1994) (based on individual industrial source processing-plant engineering specifications, emission volumes, and meteorologic information) provided exposure isopleths of sulfur dioxide concentration from each of 231 sour-gas processing-plants across the province. In contrast, a simpler measure of proximity to source(s) of varying emission rates was applied in a geographical information system based on simplified pollution decay at increasing distances from each point source. Province-wide (663,000 km(2)) surface analysis (by exposure-level classification) produced a contingency coefficient of 0.68 between the two exposure estimates. Annual exposure estimates at the 1382 dairy and 5726 beef cow-calf farms studied were highly correlated over the 10-years period (r(spearman)=0.82 and 0.83, respectively), while monthly exposure estimates were somewhat less correlated (r(spearman)=0.80 and 0.82, respectively) for the two exposure assessment methods. Crude exposure estimates from each method were similar in both direction and magnitude. PMID- 12547173 TI - Lack of associations between air emissions from sour-gas processing plants and beef cow-calf herd health and productivity in Alberta, Canada. AB - This paper describes a large-scale investigation into the effects of licensed air emissions from sour-gas processing plants on the health and productivity of beef cow-calf herds in the province of Alberta, Canada. In conjunction with a geographical information system, two atmospheric-dispersion models were used to assess historical exposures at 5726 beef cow-calf farm-sites from 1987 to 1990. We did secondary analyses of health, productivity, and management data, from a government-extension survey previously administered to beef cow-calf producers across the province. Statistical models (adjusted for potential confounding and clustering within herd and over time) were used to determine associations with estimates of sour-gas emission exposure. All analyses were conducted at the herd level. There were no significant (P>0.05) detrimental associations of exposure and the annual herd risk for culling, calf-crop delivered, calf-crop season profile, stillbirth and twinning, calfhood mortality, or calf-crop weaned. PMID- 12547174 TI - Air emissions from sour-gas processing plants and dairy-cattle reproduction in Alberta, Canada. AB - The dispersion of air pollutants from all 231 licensed sour-gas processing plants in Alberta, Canada, was modeled on a monthly basis over a 10-year period (1985 1994). Exposure estimates for sulfur dioxide (SO(2)-used as a surrogate for exposure to combusted emissions) then were assigned to 1382 provincial dairy farms using a geographical-information system. Individual average and peak exposure for periods prior to each of 15 months of age and conception (four exposure-averaging periods for each of two dispersion models) were estimated for 163,988 primiparous female dairy-cattle between 1986 and 1994. Monthly or annual average farm-site exposure estimates likewise were assigned to associated herd level data sets for the biologically relevant period of interest for each of three additional reproductive outcomes: monthly herd-average calving interval, stillbirth risk, and twinning risk. In one of the main-effects models, the maximum (i.e., peak) monthly sour-gas exposure experienced by individual-animals from birth to conception was associated with an increased time to first-calving in the very-highest exposure category (hazard ratio=0.86, 95% CI=0.80, 0.92). This equates to a decreased hazard (lambda) of calving (in each month subsequent to 22 months of age) for the highest-exposure animals (lambda=0.170) versus the zero-exposure animals (lambda=0.198) in a model with referent values for agro ecological region and season of birth. The dose-response was not consistent across the full range of exposure categories. There was significant (P=0.003) interaction of emissions with agro-ecological region. After accounting for the interaction, a more-consistent dose-response was evident for some (but not all) agro-ecological regions. This suggests that any effect of emissions on dairy heifer reproduction is subject to modification by features of soil type, vegetative cover, and/or climate. The increase in monthly herd-average calving interval on farms exposed to the very-highest levels of emissions appeared quite small and of limited practical importance within the range of expected exposures. There was no association between exposure and the risk for twinning. Herds exposed to higher emissions exhibited a slight decrease in risk for stillbirth. PMID- 12547175 TI - Epilepsy centres in Europe. Out of sight--but not out of mind? PMID- 12547176 TI - Epilepsy centres in Europe. PMID- 12547177 TI - Bethel, Bielefeld, Germany: Epilepsiezentrum Bethel. Epilepsy Centre, Bethel. PMID- 12547178 TI - Dianalund, Denmark: Kolonien Filadelfia. Dianalund Epilepsy Centre. PMID- 12547179 TI - Heemstede, The Netherlands: Stichting Epilepsie Instellingen Nederland. Foundation of Epilepsy Centres in The Netherlands. PMID- 12547180 TI - Heeze, the Netherlands: Epilepsiecentrum Kempenhaeghe. Kempenhaeghe Epilepsy Centre. PMID- 12547181 TI - Kork, Germany: Diakonie Kork Epilepsiezentrum. Epilepsy Centre, Kork. PMID- 12547182 TI - London, UK: The Chalfont Centre for Epilepsy. PMID- 12547183 TI - Sandvika, Norway: Spesialsykehuset for Epilepsi HF. The National Centre for Epilepsy at Sandvika, Norway. PMID- 12547184 TI - Tain l'Hermitageh France: Etablissement Medical de la Teppe. La Teppe Medical Center. PMID- 12547185 TI - Zurich, Switzerland: Schweizerisches Epilepsie-Zentrum. Swiss Epilepsy Centre. PMID- 12547186 TI - Functional fingerprints of folds: evidence for correlated structure-function evolution. AB - Using structural similarity clustering of protein domains: protein domain universe graph (PDUG), and a hierarchical functional annotation: gene ontology (GO) as two evolutionary lenses, we find that each structural cluster (domain fold) exhibits a distribution of functions that is unique to it. These functional distributions are functional fingerprints that are specific to characteristic structural clusters and vary from cluster to cluster. Furthermore, as structural similarity threshold for domain clustering in the PDUG is relaxed we observe an influx of earlier-diverged domains into clusters. These domains join clusters without destroying the functional fingerprint. These results can be understood in light of a divergent evolution scenario that posits correlated divergence of structural and functional traits in protein domains from one or few progenitors. PMID- 12547187 TI - Identification of a novel gene linked to parkin via a bi-directional promoter. AB - Mutations of the parkin gene on chromosome 6q25-27 are the predominant genetic cause of early-onset and autosomal recessive juvenile parkinsonism. Parkin is a multi-domain protein with ubiquitin-protein E3 ligase activity that has a role in the proteasome-mediated degradation of target substrates. Although the parkin gene contains an expanded intron/exon structure and spans more than 1.3 Mb, we have identified a novel transcript that initiates 204 bp upstream of parkin and spans over 0.6 Mb, antisense to parkin. We have tentatively named this novel gene Parkin co-regulated gene, or PACRG. A 35 bp site of bi-directional transcription activation within the common promoter was mapped using dual-luciferase assays. This region appeared to be responsible for the majority of transcription regulation of both genes, and comparison of the mouse and human sequences revealed conserved transcription factor-binding sites. A 15 bp interval within the activation region, containing a non-canonical myc-binding site, bound nuclear protein derived from human substantia nigra. Database analysis identified highly conserved homologs of PACRG encoded by the mouse and Drosophila genomes, and Northern analysis demonstrated that PACRG and parkin were co-expressed in many tissues, including brain, heart and muscle. Western analysis revealed a protein of the predicted size, approximately 30 kDa, which was expressed in mouse and human brain. Although PACRG protein lacks known functional domains, in silico prediction suggests a potential link to the ubiquitin/proteasome system. PMID- 12547188 TI - Complete nucleotide sequence of carbazole/dioxin-degrading plasmid pCAR1 in Pseudomonas resinovorans strain CA10 indicates its mosaicity and the presence of large catabolic transposon Tn4676. AB - The car and ant operons originally isolated from Pseudomonas resinovorans strain CA10 contain the genes encoding the carbazole/dioxin-degrading enzymes and anthranilate 1,2-dioxygenase, respectively, and are located on the plasmid pCAR1. The entire nucleotide sequence of pCAR1 was determined to elucidate the mechanism by which the car operon may have been assembled and distributed in nature. pCAR1 is a 199,035-bp circular plasmid, and carries 190 open reading frames. Although the incompatibility group of pCAR1 is unclear, its potential origin for replication, OriP, and Rep and Par proteins appeared to be closely related to those of plasmid pL6.5 isolated from Pseudomonas fluorescens. The potential tellurite-resistance klaABC genes identified in the neighboring region of repA gene were also related to those in IncP plasmid originally identified from pseudomonads. On the other hand, we found genes encoding proteins that showed low but significant homology (20-45% identity) with Trh and Tra proteins from Enterobacteriaceae, which are potentially involved in conjugative transfer of plasmids or genomic island, suggesting that pCAR1 is also a conjugative plasmid. In pCAR1, we found tnpAcCST genes that encoded the proteins showing >70% length wise identities with those are encoded by the toluene/xylene-degrading transposon Tn4651 of TOL plasmid pWW0. Both car and ant degradative operons were found within a 72.8-kb Tn4676 sequence defined by flanking tnpAcC and tnpST genes and bordered by a 46-bp inverted repeat (IR). Within Tn4676 and its flanking region, we found the remnants of numerous mobile genetic elements, such as the duplicated transposase genes that are highly homologous to tnpR of Tn4653 and the multiple candidates of IRs for Tn4676 and Tn4653-like element. We also found distinct regions with high and low G+C contents within Tn4676, which contain an ant operon and car operon, respectively. These results suggested that multiple step assembly could have taken place before the current structure of Tn4676 had been captured. PMID- 12547189 TI - A multifunctional turnip crinkle virus replication enhancer revealed by in vivo functional SELEX. AB - The motif1-hairpin (M1H), located on (-)-strands of Turnip Crinkle Virus (TCV) associated satellite RNA C (satC), is a replication enhancer and recombination hotspot. Results of in vivo genetic selection (SELEX: systematic evolution of ligands by exponential enrichment), where 28 bases of the M1H were randomized and then subjected to selection in plants, revealed that most winners contained one to three short motifs, many of which in their (-)-sense orientation are found in TCV and satC (-)-strand promoter elements. Ability to replicate in protoplasts correlated with fitness to accumulate in plants with one significant exception. Winner UC, containing only a seven-base replacement sequence, was the second most fit winner, yet replicated no better than a 28-base random replacement sequence. Fitness of satC containing different M1H replacement sequences could be due to enhanced satC replication or enhanced ability to affect TCV movement, since satC interferes with TCV virion accumulation, which is correlated with enhanced movement to younger tissue. Cells inoculated with TCV and UC accumulated fewer virions when compared to other winners that replicated better in protoplasts but were less fit in plants. UC, and other first and second round winners, contained structures that were on average 33% more stable in their (+)-strand orientation, and most formed hairpins with a A-rich sequence at the base. These results suggest that M1H replacement sequences contribute to the fitness of satC by either containing (-)-strand elements that enhance satRNA replication and/or a (+)-strand hairpin flanked with single-stranded sequence that enhances TCV movement. PMID- 12547190 TI - Sequence-dependent nucleosome structural and dynamic polymorphism. Potential involvement of histone H2B N-terminal tail proximal domain. AB - Relaxation of nucleosomes on an homologous series (pBR) of ca 350-370 bp DNA minicircles originating from plasmid pBR322 was recently used as a tool to study their structure and dynamics. These nucleosomes thermally fluctuated between three distinct DNA conformations within a histone N-terminal tail-modulated equilibrium: one conformation was canonical, with 1.75 turn wrapping and negatively crossed entering and exiting DNAs; another was also "closed", but with these DNAs positively crossed; and the third was "open", with a lower than 1.5 turn wrapping and uncrossed DNAs. In this work, a new minicircle series (5S) of similar size was used, which contained the 5S nucleosome positioning sequence. Results showed that DNA in pBR nucleosomes was untwisted by approximately 0.2 turn relative to 5S nucleosomes, which DNase I footprinting confirmed in revealing a approximately 1 bp untwisting at each of the two dyad-distal sites where H2B N-terminal tails pass between the two gyres. In contrast, both nucleosomes showed untwistings at the dyad-proximal sites, i.e. on the other gyre, which were also observed in the high-resolution crystal structure. 5S nucleosomes also differ with respect to their dynamics: they hardly accessed the positively crossed conformation, but had an easier access to the negatively crossed conformation. Simulation showed that such reverse effects on the conformational free energies could be simply achieved by slightly altering the trajectories of entering and exiting DNAs. We propose that this is accomplished by H2B tail untwisting at the distal sites through action at a distance ( approximately 20 bp) on H3-tail interactions with the small groove at the nucleosome entry-exit. These results may help to gain a first glimpse into the two perhaps most intriguing features of the high-resolution structure: the alignment of the grooves on the two gyres and the passage of H2B and H3 N terminal tails between them. PMID- 12547191 TI - Stepwise manipulation of DNA specificity in Flp recombinase: progressively adapting Flp to individual and combinatorial mutations in its target site. AB - The Flp protein from Saccharomyces cerevisiae is one of the site-specific tyrosine family recombinases that are used widely in genomic engineering. As a first step towards mediating directed DNA rearrangements at non-native Flp recombination targets (mFRTs), we have evolved three separate groups of Flp variants that preferentially act on mFRTs containing substitutions at the first, seventh or both positions of the Flp-binding elements. The variants that recombine the double-mutant mFRT contain a subset of the mutations present in those that are active on the single-mutant mFRTs, plus additional mutations. Specificity for and discrimination between target sites, effected primarily by amino acid residues that contact DNA, can be modulated by those that do not interact with DNA or with a DNA-contacting residue. The degree of modulation can range from relaxed DNA specificity to almost completely altered specificity. Our results suggest that combined DNA shuffling and mutagenesis of libraries of Flp variants active on distinct mFRTs can yield variants that can recombine mFRTs containing combinations of the individual mutations. PMID- 12547192 TI - Ligands differentially modulate the protein interactions of the human estrogen receptors alpha and beta. AB - The interactions of human estrogen receptor subtypes ERalpha and ERbeta with DNA and a 210 amino acid residue fragment of the coactivator protein SRC-1 bearing three nuclear receptor interaction motifs were investigated quantitatively using fluorescence anisotropy in the presence of agonist and antagonist ligands. ERalpha and ERbeta were found to bind in a similar manner to DNA, and both salt and temperature affected the affinity and/or stoichiometry of these interactions. The agonist ligands estradiol, estrone and estriol did not modify the binding of ERalpha to the fluorescein-labeled target estrogen response element. However, in the case of ERbeta, these ligands led to the formation of some higher-order protein-DNA complexes and a small decrease in affinity. The partial agonist 4 hydroxytamoxifen had little effect on either ER subtype, whereas the pure antagonist ICI 182,780 led to the cooperative formation of protein-DNA complexes of higher order than dimer, as further demonstrated by competition experiments and gel mobility-shift assays. In addition to DNA binding, the interaction of both ER subtypes with the Alexa488-labeled SRC-1 coactivator fragment was investigated by fluorescence anisotropy. The agonist ligands estrone, estradiol, estriol, genistein and ethynyl estradiol exhibited distinct capacities for inducing the recruitment of SRC-1 that were not correlated with their affinity for the receptor. Moreover, estrone and genistein exhibited subtype specificity in that they induced SRC-1 recruitment to ERbeta with much higher efficiency than in the case of ERalpha. The differential coactivator recruitment capacities of the ER agonists and their receptor subtype coactivator recruitment specificity may be linked to the molecular structure of the agonists with respect to their interactions with a specific histidine residue located at the back of the ligand binding pocket. Altogether, these quantitative in vitro studies of ER interactions reveal the complex energetic and stoichiometric consequences of changes in the chemical structures of these proteins and their ligands. PMID- 12547193 TI - Coordinated assembly of Ku and p460 subunits of the DNA-dependent protein kinase on DNA ends is necessary for XRCC4-ligase IV recruitment. AB - Repair of DNA double-strand breaks by the non-homologous end-joining pathway (NHEJ) requires a minimal set of proteins including DNA-dependent protein kinase (DNA-PK), DNA-ligase IV and XRCC4 proteins. DNA-PK comprises Ku70/Ku80 heterodimer and the kinase subunit DNA-PKcs (p460). Here, by monitoring protein assembly from human nuclear cell extracts on DNA ends in vitro, we report that recruitment to DNA ends of the XRCC4-ligase IV complex responsible for the key ligation step is strictly dependent on the assembly of both the Ku and p460 components of DNA-PK to these ends. Based on co-immunoprecipitation experiments, we conclude that interactions of Ku and p460 with components of the XRCC4-ligase IV complex are mainly DNA-dependent. In addition, under p460 kinase permissive conditions, XRCC4 is detected at DNA ends in a phosphorylated form. This phosphorylation is DNA-PK-dependent. However, phosphorylation is dispensable for XRCC4-ligase IV loading to DNA ends since stable DNA-PK/XRCC4-ligase IV/DNA complexes are recovered in the presence of the kinase inhibitor wortmannin. These findings extend the current knowledge of the assembly of NHEJ repair proteins on DNA termini and substantiate the hypothesis of a scaffolding role of DNA-PK towards other components of the NHEJ DNA repair process. PMID- 12547194 TI - TAK1 is critical for IkappaB kinase-mediated activation of the NF-kappaB pathway. AB - Cytokine treatment stimulates the IkappaB kinases, IKKalpha and IKKbeta, which phosphorylate the IkappaB proteins, leading to their degradation and activation of NF-kappaB regulated genes. A clear definition of the specific roles of IKKalpha and IKKbeta in activating the NF-kappaB pathway and the upstream kinases that regulate IKK activity remain to be elucidated. Here, we utilized small interfering RNAs (siRNAs) directed against IKKalpha, IKKbeta and the upstream regulatory kinase TAK1 in order to better define their roles in cytokine-induced activation of the NF-kappaB pathway. In contrast to previous results with mouse embryo fibroblasts lacking either IKKalpha or IKKbeta, which indicated that only IKKbeta is involved in cytokine-induced NF-kappaB activation, we found that both IKKalpha and IKKbeta were important in activating the NF-kappaB pathway. Furthermore, we found that the MAP3K TAK1, which has been implicated in IL-1 induced activation of the NF-kappaB pathway, was also critical for TNFalpha induced activation of the NF-kappaB pathway. TNFalpha activation of the NF-kappaB pathway is associated with the inducible binding of TAK1 to TRAF2 and both IKKalpha and IKKbeta. This analysis further defines the distinct in vivo roles of IKKalpha, IKKbeta and TAK1 in cytokine-induced activation of the NF-kappaB pathway. PMID- 12547195 TI - Structure of a G-quadruplex-ligand complex. AB - Stabilisation of G-quadruplex structures formed from telomeric DNA, by means of quadruplex-selective ligands, is a means of inhibiting the telomerase enzyme from catalysing the synthesis of telomeric DNA repeats. In order to understand the molecular basis of ligand-quadruplex recognition, the crystal structure has been determined of such a complex, at 1.75A resolution. This complex is between a dimeric antiparallel G-quadruplex formed from the Oxytricha nova telomeric DNA sequence d(GGGGTTTTGGGG), and a di-substituted aminoalkylamido acridine compound. The structure shows that the acridine moiety is bound at one end of the stack of G-quartets, within one of the thymine loops. It is held in place by a combination of stacking interactions and specific hydrogen bonds with thymine bases. The stability of the ligand in this binding site has been confirmed by a 2ns molecular dynamics simulation. PMID- 12547196 TI - Structure and catalytic mechanism of the cytosolic D-ribulose-5-phosphate 3 epimerase from rice. AB - Cytosolic D-ribulose-5-phosphate 3-epimerase from rice was crystallized after EDTA treatment and structurally elucidated by X-ray diffraction to 1.9A resolution. A prominent Zn(2+) site at the active center was established in a soaking experiment. The structure was compared with that of the EDTA-treated crystalline enzyme from the chloroplasts of potato plant leaves showing some structural differences, in particular the "closed" state of a strongly conserved mobile loop covering the substrate at its putative binding site. The previous proposal for the active center was confirmed and the most likely substrate binding position and conformation was derived from the locations of the bound zinc and sulfate ions and of three water molecules. Assuming that the bound zinc ion is an integral part of the enzyme, a reaction mechanism involving a well stabilized cis-enediolate intermediate is suggested. PMID- 12547197 TI - Human MYO18B, a novel unconventional myosin heavy chain expressed in striated muscles moves into the myonuclei upon differentiation. AB - We have characterized a novel unconventional myosin heavy chain, named MYO18B, that appears to be expressed mainly in human cardiac and skeletal muscles and, at lower levels, in testis. MYO18B transcript is detected in all types of striated muscles but at much lower levels compared to class II sarcomeric myosins, and it is up regulated after in vitro differentiation of myoblasts into myotubes. Phylogenetic analysis shows that this myosin belongs to the recently identified class XVIII, however, unlike the other member of this class, it seems to be unique to Vertebrate since it contains two large amino acid domains of unknown function at the N and C-termini. Immunolocalization of MYO18B protein in skeletal muscle cells shows that this myosin heavy chain is located in the cytoplasm of undifferentiated myoblasts. After in vitro differentiation into myotubes, a fraction of this protein is accumulated in a subset of myonuclei. This nuclear localization was confirmed by immunofluorescence experiments on primary cardiomyocytes and adult muscle sections. In the cytoplasm MYO18B shows a punctate staining, both in cardiac and skeletal fibers. In some cases, cardiomyocytes show a partial sarcomeric pattern of MYO18B alternating that of alpha-actinin-2. In skeletal muscle the cytoplasmic MYO18B results much more evident in the fast type fibers. PMID- 12547198 TI - Structure of a Drosophila sigma class glutathione S-transferase reveals a novel active site topography suited for lipid peroxidation products. AB - Insect glutathione-S-transferases (GSTs) are grouped in three classes, I, II and recently III; class I (Delta class) enzymes together with class III members are implicated in conferring resistance to insecticides. Class II (Sigma class) GSTs, however, are poorly characterized and their exact biological function remains elusive. Drosophila glutathione S-transferase-2 (GST-2) (DmGSTS1-1) is a class II enzyme previously found associated specifically with the insect indirect flight muscle. It was recently shown that GST-2 exhibits considerable conjugation activity for 4-hydroxynonenal (4-HNE), a lipid peroxidation product, raising the possibility that it has a major anti-oxidant role in the flight muscle. Here, we report the crystal structure of GST-2 at 1.75A resolution. The GST-2 dimer shows the canonical GST fold with glutathione (GSH) ordered in only one of the two binding sites. While the GSH-binding mode is similar to other GST structures, a distinct orientation of helix alpha6 creates a novel electrophilic substrate binding site (H-site) topography, largely flat and without a prominent hydrophobic-binding pocket, which characterizes the H-sites of other GSTs. The H site displays directionality in the distribution of charged/polar and hydrophobic residues creating a binding surface that explains the selectivity for amphipolar peroxidation products, with the polar-binding region formed by residues Y208, Y153 and R145 and the hydrophobic-binding region by residues V57, A59, Y211 and the C-terminal V249. A structure-based model of 4-HNE binding is presented. The model suggest that residues Y208, R145 and possibly Y153 may be key residues involved in catalysis. PMID- 12547199 TI - Deriving topological constraints from functional data for the design of reagentless fluorescent immunosensors. AB - The possibility of obtaining, from any antibody, a fluorescent conjugate which responds to the binding of the antigen by a variation of fluorescence, would be of great interest in the micro- and nano-analytical sciences. This possibility was explored with antibody mAb4E11, which is directed against the dengue virus and for which no structural data is available. Three rules of design were developed to identify residues of the antibody to which a fluorophore could be chemically coupled, after changing them to cysteine by mutagenesis. (i) The target residue belonged to the hypervariable loops of the antibody. (ii) It was adjacent, along the amino acid sequence of the antibody, to a residue which was functionally important for the interaction with the antigen. (iii) It was not important in itself for the interaction with the antigen. Eight conjugates between a single chain variable fragment of mAb4E11 and an environment-sensitive fluorophore were constructed. Three of them showed an increase in their fluorescence intensity by 1.5-2.8-fold on antigen binding, without loss of affinity. This increase allowed the titration of the antigen in serum above a threshold concentration of 10nM. Experiments of quenching with potassium iodide suggested that the fluorescence variation was due to a shielding of the fluorescent group from the solvent by the binding of the antigen, and that therefore its mechanism is general. PMID- 12547200 TI - Three-dimensional structure and substrate binding of Bacillus stearothermophilus neopullulanase. AB - Crystal structures of Bacillus stearothermophilus TRS40 neopullulanase and its complexes with panose, maltotetraose and isopanose were determined at resolutions of 1.9, 2.4, 2.8 and 3.2A, respectively. Since the latter two carbohydrates are substrates of this enzyme, a deactivated mutant at the catalytic residue Glu357- >Gln was used for complex crystallization. The structures were refined at accuracies with r.m.s. deviations of bond lengths and bond angles ranging from 0.005A to 0.008A and 1.3 degrees to 1.4 degrees, respectively. The active enzyme forms a dimer in the crystalline state and in solution. The monomer enzyme is composed of four domains, N, A, B and C, and has a (beta/alpha)(8)-barrel in domain A. The active site lies between domain A and domain N from the other monomer. The results show that dimer formation makes the active-site cleft narrower than those of ordinary alpha-amylases, which may contribute to the unique substrate specificity of this enzyme toward both alpha-1,4 and alpha-1,6 glucosidic linkages. This specificity may be influenced by the subsite structure. Only subsites -1 and -2 are commonly occupied by the product and substrates, suggesting that equivocal recognition occurs at the other subsites, which contributes to the wide substrate specificity of this enzyme. PMID- 12547201 TI - Solution structure of the Bacillus subtilis T-box antiterminator RNA: seven nucleotide bulge characterized by stacking and flexibility. AB - The T-box transcription antitermination regulatory system is an important mechanism for regulation of expression of aminoacyl-tRNA synthetase, amino acid biosynthesis and transporter gene expression in Gram-positive bacteria. Antitermination is dependent on a complex set of interactions between uncharged tRNA and the leader region of the mRNA of the regulated gene. Here, we report the solution structure of a model RNA, based on the Bacillus subtilis tyrS antiterminator, determined to an rmsd of 3.47A for all nine converged structures and 2.66A for the seven structures representing the consensus family. The antiterminator is comprised of two short helices with an intervening 7nt bulge. The bulge region of the antiterminator, which ultimately interacts with the acceptor end of tRNA, exhibits extensive stacking at the 3' end (encompassing the highly conserved ACC residues) and is the site of a pronounced kink between the two flanking helices. The 5' end of the bulge exhibits evidence of conformational flexibility. On the basis of the structural studies, there is no indication that the bases at the 5' end of the bulge that ultimately base-pair with tRNA are pre organized for binding. Instead, the data are consistent with a model in which the stacking-induced structure at the 3' end of the bulge may facilitate the pre selection of a set of conformations for the tRNA to sample during binding. PMID- 12547202 TI - Aspartate transcarbamylase from the hyperthermophilic archaeon Pyrococcus abyssi: thermostability and 1.8A resolution crystal structure of the catalytic subunit complexed with the bisubstrate analogue N-phosphonacetyl-L-aspartate. AB - The Pyrococcus abyssi aspartate transcarbamylase (ATCase) shows a high degree of structural conservation with respect to the well-studied mesophilic Escherichia coli ATCase, including the association of catalytic and regulatory subunits. The adaptation of its catalytic function to high temperature was investigated, using enzyme purified from recombinant E.coli cells. At 90 degrees C, the activity of the trimeric catalytic subunit was shown to be intrinsically thermostable. Significant extrinsic stabilization by phosphate, a product of the reaction, was observed when the temperature was raised to 98 degrees C. Comparison with the holoenzyme showed that association with regulatory subunits further increases thermostability. To provide further insight into the mechanisms of its adaptation to high temperature, the crystal structure of the catalytic subunit liganded with the analogue N-phosphonacetyl-L-aspartate (PALA) was solved to 1.8A resolution and compared to that of the PALA-liganded catalytic subunit from E.coli. Interactions with PALA are strictly conserved. This, together with the similar activation energies calculated for the two proteins, suggests that the reaction mechanism of the P.abyssi catalytic subunit is similar to that of the E.coli subunit. Several structural elements potentially contributing to thermostability were identified: (i) a marked decrease in the number of thermolabile residues; (ii) an increased number of charged residues and a concomitant increase of salt links at the interface between the monomers, as well as the formation of an ion pair network at the protein surface; (iii) the shortening of three loops and the shortening of the N and C termini. Other known thermostabilizing devices such as increased packing density or reduction of cavity volumes do not appear to contribute to the high thermostability of the P.abyssi enzyme. PMID- 12547203 TI - Solution structure of the R3H domain from human Smubp-2. AB - The R3H domain is a conserved sequence motif, identified in over 100 proteins, that is thought to be involved in polynucleotide-binding, including DNA, RNA and single-stranded DNA. In this work the 3D structure of the R3H domain from human Smubp-2 was determined by NMR spectroscopy. It is the first 3D structure determination of an R3H domain. The fold presents a small motif, consisting of a three-stranded antiparallel beta-sheet and two alpha-helices, which is related to the structures of the YhhP protein and the C-terminal domain of the translational initiation factor IF3. The similarities are non-trivial, as the amino acid identities are below 10%. Three conserved basic residues cluster on the same face of the R3H domain and could play a role in nucleic acid recognition. An extended hydrophobic area at a different site of the molecular surface could act as a protein-binding site. A strong correlation between conservation of hydrophobic amino acids and side-chain solvent protection indicates that the structure of the Smubp-2 R3H domain is representative of R3H domains in general. PMID- 12547204 TI - NMR structure of a variant human prion protein with two disulfide bridges. AB - The nuclear magnetic resonance structure of the globular domain with residues 121 230 of a variant human prion protein with two disulfide bonds, hPrP(M166C/E221C), shows the same global fold as wild-type hPrP(121-230). It contains three alpha helices of residues 144-154, 173-194 and 200-228, an anti-parallel beta-sheet of residues 128-131 and 161-164, and the disulfides Cys166-Cys221 and Cys179-Cys214. The engineered extra disulfide bond in the presumed "protein X"-binding site is accommodated with slight, strictly localized conformational changes. High compatibility of hPrP with insertion of a second disulfide bridge in the protein X epitope was further substantiated by model calculations with additional variant structures. The ease with which the hPrP structure can accommodate a variety of locations for a second disulfide bond within the presumed protein X-binding epitope suggests a functional role for the extensive perturbation by a natural second disulfide bond of the corresponding region in the human doppel protein. PMID- 12547205 TI - Kinetic mechanism and catalysis of a native-state prolyl isomerization reaction. AB - Folding of tendamistat is a rapid two-state process for the majority of the unfolded molecules. In fluorescence-monitored refolding kinetics about 8% of the unfolded molecules fold slowly (lambda=0.083s(-1)), limited by peptidyl-prolyl cis-trans isomerization. This is significantly less than expected from the presence of three trans prolyl-peptide bonds in the native state. In interrupted refolding experiments we detected an additional very slow folding reaction (lambda=0.008s(-1) at pH 2) with an amplitude of about 12%. This reaction is caused by the interconversion of a highly structured intermediate to native tendamistat. The intermediate has essentially native spectroscopic properties and about 2% of it remain populated in equilibrium after folding is complete. Catalysis by human cyclophilin 18 identifies this very slow reaction as a prolyl isomerization reaction. This shows that prolyl-isomerases are able to efficiently catalyze native state isomerization reactions, which allows them to influence biologically important regulatory conformational transitions. Folding kinetics of the proline variants P7A, P9A, P50A and P7A/P9A show that the very slow reaction is due to isomerization of the Glu6-Pro7 and Ala8-Pro9 peptide bonds, which are located in a region that makes strong backbone and side-chain interactions to both beta-sheets. In the P50A variant the very slow isomerization reaction is still present but native state heterogeneity is not observed any more, indicating a long-range destabilizing effect on the alternative native state relative to N. These results enable us to include all prolyl and non-prolyl peptide bond isomerization reactions in the folding mechanism of tendamistat and to characterize the kinetic mechanism and the energetics of a native-state prolyl isomerization reaction. PMID- 12547206 TI - Cooperativity, smooth energy landscapes and the origins of topology-dependent protein folding rates. AB - The relative folding rates of simple, single-domain proteins, proteins whose folding energy landscapes are smooth, are highly dispersed and strongly correlated with native-state topology. In contrast, the relative folding rates of small, Go-potential lattice polymers, which also exhibit smooth energy landscapes, are poorly dispersed and insignificantly correlated with native-state topology. Here, we investigate this discrepancy in light of a recent, quantitative theory of two-state folding kinetics, the topomer search model. This model stipulates that the topology-dependence of two-state folding rates is a direct consequence of the extraordinarily cooperative equilibrium folding of simple proteins. We demonstrate that traditional Go polymers lack the extreme cooperativity that characterizes the folding of naturally occurring, two-state proteins and confirm that the folding rates of a diverse set of Go 27-mers are poorly dispersed and effectively uncorrelated with native state topology. Upon modestly increasing the cooperativity of the Go-potential, however, significantly increased dispersion and strongly topology-dependent kinetics are observed. These results support previous arguments that the cooperative folding of simple, single domain proteins gives rise to their topology-dependent folding rates. We speculate that this cooperativity, and thus, indirectly, the topology-rate relationship, may have arisen in order to generate the smooth energetic landscapes upon which rapid folding can occur. PMID- 12547207 TI - An accurate, sensitive, and scalable method to identify functional sites in protein structures. AB - Functional sites determine the activity and interactions of proteins and as such constitute the targets of most drugs. However, the exponential growth of sequence and structure data far exceeds the ability of experimental techniques to identify their locations and key amino acids. To fill this gap we developed a computational Evolutionary Trace method that ranks the evolutionary importance of amino acids in protein sequences. Studies show that the best-ranked residues form fewer and larger structural clusters than expected by chance and overlap with functional sites, but until now the significance of this overlap has remained qualitative. Here, we use 86 diverse protein structures, including 20 determined by the structural genomics initiative, to show that this overlap is a recurrent and statistically significant feature. An automated ET correctly identifies seven of ten functional sites by the least favorable statistical measure, and nine of ten by the most favorable one. These results quantitatively demonstrate that a large fraction of functional sites in the proteome may be accurately identified from sequence and structure. This should help focus structure-function studies, rational drug design, protein engineering, and functional annotation to the relevant regions of a protein. PMID- 12547208 TI - High concentrations of viscogens decrease the protein folding rate constant by prematurely collapsing the coil. AB - In several studies, viscogenic osmolytes have been suggested to decrease the folding rate constant of polypeptides by slowing their motion through the solvent. Here, we show that osmolytes may slow protein folding by prematurely collapsing the coil. At low or moderate concentrations of osmolytes (<30%), folding of the two-state protein CI2 becomes faster with increasing osmolyte concentrations, suggesting that the kinetics are governed by protein stability. However, at higher concentrations of osmolyte, the coil collapses in the dead time of the refolding experiment, causing a dramatic drop in the folding rate. The collapsed state is non-native and appears to be different for different osmolytes. PMID- 12547209 TI - Sequence and structure patterns in proteins from an analysis of the shortest helices: implications for helix nucleation. AB - The shortest helices (three-length 3(10) and four-length alpha), most abundant among helices of different lengths, have been analyzed from a database of protein structures. A characteristic feature of three-length 3(10)-helices is the shifted backbone conformation for the C-terminal residue (phi,psi angles: -95 degrees,0 degrees ), compared to the rest of the helix (-62 degrees,-24 degrees ). The deviation can be attributed to the release of electrostatic repulsion between the carbonyl oxygen atoms at the two C-terminal residues and further stabilization (due to a more linear geometry) of an intrahelical hydrogen bond. A consequence of this non-canonical C-terminal backbone conformation can be a potential origin of helix kinks when a 3(10)-helix is sequence-contiguous at the alpha-helix N terminal. An analysis of hydrogen bonding, as well as hydrophobic interactions in the shortest helices shows that capping interactions, some of them not observed for longer helices, dominate at the N termini. Further, consideration of the distribution of amino acid residues indicates that the shortest helices resemble the N-terminal end of alpha-helices rather than the C terminus, implying that the folding of helices may be initiated at the N-terminal end, which does not get propagated in the case of the shortest helices. Finally, pairwise comparison of beta-turns and the shortest helices, based on correlation matrices of site specific amino acid composition, and the relative abundance of these short secondary structural elements, leads to a helix nucleation scheme that considers the formation of an isolated beta-turn (and not an alpha-turn) as the helix nucleation step, with shortest 3(10)-helices as intermediates between the shortest alpha-helix and the beta-turn. Our results ascribe an important role played by shortest 3(10)-helices in proteins with important structural and folding implications. PMID- 12547210 TI - Structural analysis of the rate-limiting transition states in the folding of Im7 and Im9: similarities and differences in the folding of homologous proteins. AB - The bacterial immunity proteins Im7 and Im9 fold with mechanisms of different kinetic complexity. Whilst Im9 folds in a two-state transition at pH 7.0 and 10 degrees C, Im7 populates an on-pathway intermediate under these conditions. In order to assess the role of sequence versus topology in the folding of these proteins, and to analyse the effect of populating an intermediate on the landscape for folding, we have determined the conformational properties of the rate-limiting transition state for Im9 folding/unfolding using Phi(F)-value analysis and have compared the results with similar data obtained previously for Im7. The data show that the rate-limiting transition states for Im9 and Im7 folding/unfolding are similar: both are compact (beta(T)=0.94 and 0.89, respectively) and contain three of the four native helices docked around a specific hydrophobic core. Significant differences are observed, however, in the magnitude of the Phi(F)-values obtained for the two proteins. Of the 20 residues studied in both proteins, ten have Phi(F)-values in Im7 that exceed those in Im9 by more than 0.2, and of these five differ by more than 0.4. The data suggest that the population of an intermediate in Im7 results in folding via a transition state ensemble that is conformationally restricted relative to that of Im9. The data are consistent with the view that topology is an important determinant of folding. Importantly, however, they also demonstrate that while the folding transition state may be conserved in homologous proteins that fold with two and three-state kinetics, the population of an intermediate can have a significant effect on the breadth of the transition state ensemble. PMID- 12547211 TI - Detergent structure in crystals of the integral membrane light-harvesting complex LH2 from Rhodopseudomonas acidophila strain 10050. AB - Integral membrane proteins are solubilized by their incorporation into a detergent micelle. The detergent micelle has a critical influence on the formation of a three-dimensional crystal lattice. The bulk detergent phase is not seen in X-ray crystal structures of integral membrane proteins, due to its disordered character. Here, we describe the detergent structure present in crystals of the peripheral light-harvesting complex of the purple bacteria Rhodopseudomonas acidophila strain 10050 at a maximal resolution of 12A as determined by neutron crystallography. The LH2 molecule has a toroidal shape and spans the membrane completely in vivo. A volume of 16% of the unit cell could be ascribed to detergent tails, localized on both the inner and outer hydrophobic surfaces of the molecule. The detergent tail volumes were found to be associated with individual LH2 molecules and had no direct role in the formation of the crystalline lattice. PMID- 12547212 TI - COMPASS: a tool for comparison of multiple protein alignments with assessment of statistical significance. AB - We present a novel method for the comparison of multiple protein alignments with assessment of statistical significance (COMPASS). The method derives numerical profiles from alignments, constructs optimal local profile-profile alignments and analytically estimates E-values for the detected similarities. The scoring system and E-value calculation are based on a generalization of the PSI-BLAST approach to profile-sequence comparison, which is adapted for the profile-profile case. Tested along with existing methods for profile-sequence (PSI-BLAST) and profile profile (prof_sim) comparison, COMPASS shows increased abilities for sensitive and selective detection of remote sequence similarities, as well as improved quality of local alignments. The method allows prediction of relationships between protein families in the PFAM database beyond the range of conventional methods. Two predicted relations with high significance are similarities between various Rossmann-type folds and between various helix-turn-helix-containing families. The potential value of COMPASS for structure/function predictions is illustrated by the detection of an intricate homology between the DNA-binding domain of the CTF/NFI family and the MH1 domain of the Smad family. PMID- 12547214 TI - Increased duodenal DMT-1 expression and unchanged HFE mRNA levels in HFE associated hereditary hemochromatosis and iron deficiency. AB - HFE-associated hereditary hemochromatosis is characterized by imbalances of iron homeostasis and alterations in intestinal iron absorption. The identification of the HFE gene and the apical iron transporter divalent metal transporter-1, DMT-1, provide a direct method to address the mechanisms of iron overload in this disease. The aim of this study was to evaluate the regulation of duodenal HFE and DMT-1 gene expression in HFE-associated hereditary hemochromatosis. Small bowel biopsies and serum iron indices were obtained from a total of 33 patients. The study population comprised 13 patients with hereditary hemochromatosis (C282Y homozygous), 10 patients with iron deficiency anemia, and 10 apparently healthy controls, all of whom were genotyped for the two common mutations in the HFE gene (C282Y and H63D). Total RNA was isolated from tissue and amplified via RT-PCR for HFE, DMT-1, and the internal control GAPDH. DMT-1 protein expression was additionally assessed by immunohistochemistry. Levels of HFE mRNA did not differ significantly between patient groups (P = 0.09), specifically between C282Y homozygotes and iron deficiency anemic patients, when compared to controls (P = 0.09, P = 0.9, respectively). In contrast, DMT-1 mRNA levels were at least twofold greater in patients with hereditary hemochromatosis and iron deficiency anemia when compared to controls (P = 0.02, P = 0.01, respectively). Heightened DMT-1 protein expression correlated with mRNA levels in all patients. Loss of HFE function in hereditary hemochromatosis is not derived from inhibition of its gene expression. DMT-1 expression in C282Y homozygote subjects is consistent with the hypothesis of a "paradoxical" duodenal iron deficiency in hereditary hemochromatosis. The observed twofold upregulation of the DMT-1 is consistent with the slow but steady increase in body iron stores observed in those presenting with clinical features of hereditary hemochromatosis. PMID- 12547215 TI - Rat erythrocytes express the anionic amino acid transport protein EAAC1. AB - EAAC1 is a widely expressed protein which transports anionic amino acids in a Na(+)-dependent fashion. Rat erythrocytes have generally been thought to be impermeant to anionic amino acids. Utilizing immunoelectron microscopy, we have demonstrated the presence of EAAC1-immunoreactive protein within rat erythrocytes. Immunoblotting revealed the presence of an approximately 60-kDa protein, consistent with EAAC1, in erythrocyte membranes. Specificity was confirmed by peptide competition. In conclusion, EAAC1 is expressed in rat erythrocytes. PMID- 12547216 TI - Frequency of the HFE gene mutations in five Italian populations. AB - Genetic hemochromatosis is an autosomal recessive disorder characterized by iron overload and a variety of clinical manifestations such as liver cirrhosis and arthropathy. It is the most common genetic disease of northern European populations. The principal gene responsible for hereditary hemochromatosis, designated HFE, is located on chromosome 6 in the HLA region. The single point mutation 845A, changing cysteine at position 282 to tyrosine (C282Y), in this gene has been identified as the main genetic basis of hereditary hemochromatosis. Two other mutations, 187G, a histidine to aspartate at amino acid 63 (H63D), and 193T, a serine to cysteine at amino acid 65 (S65C), appear to be associated with milder forms of hereditary hemochromatosis. There is a high prevalence of the C282Y mutation in northern European populations, whereas in those of the Mediterranean basin the prevalence seems low and almost absent in Far East countries. This mutation seems usually to occur on the ancestral haplotype 7.1. Accordingly, a Celtic origin of this mutation has been suggested. The aim of this study was to determine the frequency of HFE gene mutations in five geographic regions in Italy. Samples were tested for C282Y, H63D, and S65C mutations of the HFE gene according to methods of each laboratory and the results were standardized with the exchange of typed samples between the different laboratories. In addition, C282Y-positive DNA samples were typed for D6S105 allele 8 and HLA-A3 by ARMS-PCR. We have found that the allele frequency of the C282Y mutation decreases from northeast Italy (Friuli, 6%) to northwest Italy (Piedmont, 4.8%) and to central Italy (Emilia-Romagna, 1.7%). However, this mutation is lacking in the two regions of the Mediterranean basin's center (Sicily and Sardinia). Accordingly, a significant difference in the frequency of the mutation was observed between these Italian regions (P = 0.07 x 10(-3)). In contrast, no difference was observed in allele frequency of H63D in the five Italian regions. Finally, as regards the S65C mutation a very low frequency was observed in Friuli, Emilia-Romagna, and Sardinia, whereas in Sicily and Piedmont we have not found this mutation. In conclusion, these data are consistent with the hypothesis that the C282Y mutation occurred in Caucasian populations of Celtic origin, whereas the H63D mutation is more ancient as demonstrated by the ubiquitous distribution. PMID- 12547217 TI - Hemochromatosis protein (HFE) and tumor necrosis factor receptor 2 (TNFR2) influence tissue iron levels: elements of a common gut pathway? AB - Quantitative genetic analysis of hepatic and splenic iron levels in recombinant inbred mice yielded a quantitative trait locus that was found to coincide with the genomic locale encompassing the tumor necrosis factor receptor 2 gene (Tnfr2). When fed an iron-enriched diet, mice nullizygous with respect to Tnfr2, but not the Tnfr1 gene, showed a significant increase in splenic non-heme iron levels. This result contrasted with mice deficient in the hemochromatosis protein, HFE, which demonstrated a significant increase in normally high hepatic iron levels, but no change in splenic iron, when fed an iron-enriched chow. Both Tnfr2 knockout and HFE knockout mice fed an iron-enriched diet failed to demonstrate intestinal epithelial cell iron following the application of the Perls' stain, as compared to both Tnfr1 knockout and normal control mice. Moreover, intestinal intraepithelial lymphocytes (IELs) isolated from HFE knockout mice did not show an increase in TNF expression following challenge with the iron-enriched diet, in contrast to normal controls. These results suggest that HFE and TNFR2 are both involved in regulating iron deposition in tissues and that the regulation occurs at the level of the intestine through IEL-orchestrated production of TNF following the binding to TNFR2. These data suggest that HFE and TNFR2 may contribute to a common pathway of the iron stores regulator insuring the controlled efflux of gut iron. PMID- 12547218 TI - Genetic analysis of hemostasis and thrombosis using vascular occlusion. AB - The zebrafish is an excellent model for mammalian hemostasis and thrombosis since it possesses coagulation factors, thrombocyte receptors and responds to anti coagulant and anti-platelet drugs commonly used in clinical treatment. In this study, exposure of larvae to FeCl(3) or laser irradiation produced a vessel injury that caused a visible vascular occlusion as a result of thrombus formation. Using the time to vascular occlusion as an assay, two screening strategies were tested for their utility in identifying novel genes involved in thrombosis. Morpholino knockdown studies of zebrafish factor VII showed a prolongation of the time to occlusion of the vessel whereas knockdown of the recently discovered factor VIIi resulted in a shortening of the time. Genetic screening of a population of zebrafish identified mutants that showed a prolongation of the time to occlusion. Bulk segregant analysis showed linkage of one mutant to a locus, victoria, on linkage group 7. Thus, the vascular occlusion assay developed in this report measures in vivo thrombus formation and is a powerful tool for identifying novel genes involved in thrombosis. PMID- 12547220 TI - History of iron in medicine. PMID- 12547221 TI - Post-transcriptional regulation of human iron metabolism by iron regulatory proteins. AB - In mammalian iron metabolism, ferritin, transferrin receptor and several other iron metabolism genes are post-transcriptionally regulated. Iron regulatory proteins 1 and 2 are cytosolic proteins that bind to RNA stem-loops known as iron responsive elements in several transcripts. We have studied the role of these proteins in knockout mice and discovered that misregulation of iron metabolism can be a primary cause of neurodegeneration. PMID- 12547222 TI - Role of the ferroportin iron-responsive element in iron and nitric oxide dependent gene regulation. AB - The newly described iron transporter, ferroportin (MTP1, IREG1), is expressed in a variety of tissues including the duodenum and cells of the mononuclear phagocyte system (MPS). In the MPS, ferroportin is hypothesized to be a major exporter of iron scavenged from senescent erythrocytes. Changes in iron metabolism, including the sequestration of iron in the MPS, are characteristic of both acute and chronic inflammation and these conditions induce changes in ferroportin expression. In a mouse model of acute inflammation, LPS administration is associated with reduced MPS ferroportin protein and mRNA expression. In addition, the ferroportin 5' UTR also has an iron-responsive element that binds to the iron-response proteins, but whether there is a role for this IRE in inflammation induced regulation of ferroportin has been unclear. A luciferase reporter gene under the control of the mouse ferroportin promoter and 5' UTR was used to determine if this 5' UTR conferred IRE-dependent regulation on this reporter gene. Stimulation of reporter gene transfected RAW 264.7 cells (a mouse macrophage cell line) with LPS resulted in IRE-dependent inhibition of luciferase production. Inhibitors of nitric oxide synthase abrogated the IRE dependent effect of LPS. In addition, direct treatment of RAW 264.7 and with NO donor S-nitroso-N-acetylpenicillamine resulted in IRE-dependent down-regulation of luciferase expression. The effect of NO was consistent with IRP1/IRE mediated translation block. There are most likely both inflammation-mediated transcriptional and post-transcriptional (IRE-dependent) mechanisms for inhibiting ferroportin expression in MPS cells. PMID- 12547223 TI - Hepcidin, a new iron regulatory peptide. AB - Maintaining normal iron homeostasis is essential for the organism, as both iron deficiency and iron excess are associated with cellular dysfunction. Recently, several lines of evidence have suggested that hepcidin, a peptide mainly produced by the liver, plays a major role in the control of body iron homeostasis. The subject of this paper is to summarize the advances toward the understanding of function and regulation of hepcidin in iron metabolism and to provide new data on the regulation of hepcidin gene expression by erythropoietin, the major regulator of mammalian erythropoiesis. PMID- 12547224 TI - Pathways of iron absorption. AB - Iron is vital for all living organisms but excess iron can be lethal because it facilitates free radical formation. Thus iron absorption is carefully regulated to maintain an equilibrium between absorption and body loss of iron. In countries where meat is a significant part of the diet, most body iron is derived from dietary heme because heme binds few of the dietary chelators that bind inorganic iron. Uptake of heme into enterocytes occurs as a metalloporphyrin in an endosomal process. Intracellular iron is released from heme by heme oxygenase to enter plasma as inorganic iron. Ferric iron is absorbed via a beta(3) integrin and mobilferrin pathway (IMP) which is unshared with other nutritional metals. Ferrous iron uptake is facilitated by a DMT-1 pathway which is shared with manganese. In the iron deficient gut, large quantities of both mobilferrin and DMT-1 are found in goblet cells and intraluminal mucins suggesting that they are secreted with mucin into the intestinal lumen to bind iron to facilitate uptake by the cells. In the cytoplasm, IMP and DMT associate in a large protein complex called paraferritin which serves as a ferrireductase. Paraferritin solublizes iron binding proteins and reduces iron to make iron available for production of iron containing proteins such as heme. Iron uptake by intestinal absorptive cells is regulated by the iron concentration within the cell. Except in hemochromatosis it remains in equilibrium with total body stores via transferrin receptors on the basolateral membrane of absorptive cells. Increased intracellular iron either up regulates or satiates iron binding proteins on regulatory proteins to alter their location in the intestinal mucosa. PMID- 12547225 TI - Molecular and functional roles of duodenal cytochrome B (Dcytb) in iron metabolism. AB - Dcytb has been identified as the mammalian transplasma ferric reductase that catalyzes the reduction of ferric to ferrous iron in the process of iron absorption. Its mRNA and protein levels are up-regulated by several independent stimulators of iron absorption. Furthermore, its cDNA encodes putative binding sites for heme and ascorbic acid. Using Northern and Western blots, RT-PCR and confocal microscopy, we studied the expression and localisation of Dcytb in cell lines and tissues of CD1 mice. Dcytb expression and function were modulated by iron. Dcytb and DMT1, both predominantly localised in the apical region of the duodenum were up-regulated in iron deficiency. Dcytb, the iron regulated ferric reductase may also utilize cytoplasmic ascorbate as electron donor for transmembrane reduction of iron. Dcytb expression was found in other tissues apart from the duodenum and its regulation and functions at these other sites are of interest in iron metabolism. PMID- 12547226 TI - Decreased liver hepcidin expression in the Hfe knockout mouse. AB - Hepcidin is a circulating antimicrobial peptide which has been proposed to regulate the uptake of dietary iron and its storage in reticuloendothelial macrophages. Transgenic mice lacking hepcidin expression demonstrate abnormalities of iron homeostasis similar to Hfe knockout mice and to patients with HFE-associated hereditary hemochromatosis (HH). To identify any association between liver hepcidin expression and the iron homeostasis abnormalities observed in HH, we compared liver hepcidin mRNA content in wild type and Hfe knockout mice. Because the iron homeostasis abnormalities in the Hfe knockout mice are greatest early in life, we analyzed mice at different ages. At four weeks of age, Hfe knockout mice had significantly decreased liver hepcidin mRNA expression compared to wild type mice. The decreased hepcidin expression was associated with hepatic iron deposition, elevated transferrin saturations, and decreased splenic iron concentrations. At 10 weeks of age, despite marked hepatic iron loading, Hfe knockout mice demonstrated liver hepcidin mRNA expression similar to that observed in wild type mice. Placing 8 week-old wild type and Hfe knockout mice on a 2% carbonyl iron diet for 2 weeks led to a similar degree of hepatic iron loading in each group. However, while the wild type mice demonstrated a mean five fold increase in liver hepcidin mRNA, no change was observed in the Hfe knockout mice. The lack of an increase in liver hepcidin expression in these iron-loaded Hfe knockout mice was associated with sparing of iron deposition into the spleen. These data indicate that the normal relationship between body iron stores and liver hepcidin mRNA levels is altered in Hfe knockout mice, such that liver hepcidin expression is relatively decreased. We speculate that decreased hepcidin expression relative to body iron stores contributes to the iron homeostasis abnormalities characteristic of HH. PMID- 12547227 TI - The ceruloplasmin homolog hephaestin and the control of intestinal iron absorption. AB - Hephaestin is the gene affected in the sex-linked anemic (sla) mouse. These animals have a defect in the export of iron from intestinal enterocytes into the circulation and this implicates hephaestin in the basolateral transfer step of iron absorption. Hephaestin is homologous to the plasma copper-containing protein ceruloplasmin, and all residues involved in copper binding and disulfide bond formation in ceruloplasmin are conserved in hephaestin. Unlike ceruloplasmin, hephaestin is an integral membrane protein with a single trans-membrane domain. It is highly expressed throughout the small intestine, to a lesser extent in the colon, and at low levels in several other tissues. Surprisingly, most hephaestin appears to be located intracellularly in a perinuclear distribution. Like ceruloplasmin, hephaestin has a ferroxidase activity which is predicted to underlie its biological function. In addition, its expression is stimulated under iron deficient conditions. Analysis of the sla mouse has supported our model for the regulation of intestinal iron absorption whereby changes in systemic iron requirements alter the levels of basolateral transport components with subsequent regulation of brush border transport. PMID- 12547228 TI - Mitochondrial ferritin: a new player in iron metabolism. AB - Mitochondrial ferritin (MtF) is a novel H-type ferritin encoded by an intronless gene on chromosome 5q23.1. The protein is synthesized as a precursor of about 30 kDa that is targeted to mitochondria by a leader sequence of 60 amino acids. This leader is proteolytically removed inside the mitochondria and the resulting 22 kDa subunit forms typical ferritin shells. These shells have ferroxidase activity and are therefore likely to sequester potentially harmful free iron. However, this may be a limited function since MtF has a very restricted tissue expression. High amounts are found in testis but only very low levels are found in iron storage organs. The levels of MtF appear to correlate more with mitochondrial abundance than with iron metabolism. MtF does not seem to be an obligatory intermediate in transfer of free iron to heme and other iron compounds in mitochondria. However, its level increases dramatically in sideroblastic anemia when heme synthesis is disrupted. This increased synthesis does not appear to involve the classical translational control since MtF mRNA lacks an apparent iron response element. In transfected HeLa cells added iron is incorporated as quickly into MtF as into cytosolic ferritin. In addition, increased levels of MtF cause a redistribution of iron from cytosol to mitochondria and this effect is enhanced by iron chelation. Thus high levels of MtF result in an iron deficient phenotype in cytosol with decreased expression of ferritin and increased expression of transferrin receptor. This avidity for iron may explain why MtF levels are maintained at low levels in most normal cells. The regulation of MtF expression and possible therapeutic applications of MtF in neurological disorders involving increased iron deposition are topics for future research. PMID- 12547229 TI - Mechanisms and regulation of intestinal iron absorption. AB - Iron absorption from the small intestine is regulated according to the body's needs, increasing in iron deficiency and decreasing in iron overload. It has been proposed that the efficiency of absorption is determined by the amount of iron acquired by developing enterocytes when they are in the crypts of Lieberkuhn and that this regulates expression of iron transporters such as DMT1 in mature enterocytes of the intestinal villi. In the crypts the cells take up iron from plasma transferrin by receptor-mediated endocytosis, a process that is influenced by the hemochromatosis protein, HFE. Hence, the availability of plasma transferrin-bound iron and the expression and function of transferrin receptors (TfR1), HFE and DMT1 should all contribute to the absorptive capacity of villus enterocytes. These aspects of the regulation and mechanism of iron absorption were investigated in genetically normal rats and mice, and in Belgrade anemic (b/b) rats and HFE knockout mice. In most experiments the function of the TfR1 was assessed by the uptake of radiolabeled transferrin-bound iron given intravenously. Absorption of non-heme iron was measured using closed in situ duodenal loops. The expression and cellular distribution of DMT1 and TfR1 were determined by in situ hybridisation and immunohistochemistry. The uptake of transferrin-bound iron and expression of functional TfR1 was shown to occur mainly in crypt cells and to be proportional to the plasma concentration of iron. It was not impaired by the mutation of DMT1 that occurs in b/b rats but was impaired in HFE knockout mice. Iron absorption was increased in these mice but was still influenced by the level of iron stores, as in normal mice. These results are in accordance with the proposed regulation of iron absorption and suggest that DMT1 is not the only iron transporter operating within endosomes of crypt cells. This view was supported by the failure to detect DMT1 mRNA or protein in crypt cells. Expression of DMT1 mRNA and protein started at the crypt villus junction and increased to reach highest levels in the mid-villus region. Greater expression was found in iron deficiency and less in iron loaded animals than in controls and in the iron deficient rats most of the protein was present on the brush border membrane. In normal rats the efficiency of iron absorption parallelled the level of DMT1 expression, but in b/b rats absorption was very low and independent of dietary iron content even though DMT1 was present in villus enterocytes. The results confirm the essential role of DMT1 in the uptake phase of non-heme iron absorption. When normal rats previously fed a low iron diet were given a bolus of iron by stomach tube, the subsequent absorption of iron from a test dose placed in the duodenum diminished in parallel with the expression of DMT1 mRNA and protein, commencing within 1hour and reaching low levels by 7 hours. The margination of DMT1 to the brush border membrane disappeared. These results show the level of expression and intracellular distribution and function of DMT1 respond very quickly to the iron content of the diet as well as being affected by storage iron levels. PMID- 12547230 TI - Nitric oxide-mediated modulation of iron regulatory proteins: implication for cellular iron homeostasis. AB - Iron regulatory proteins (IRP1 and IRP2) control the synthesis of transferrin receptors (TfR) and ferritin by binding to iron-responsive elements (IREs) that are located in the 3' untranslated region (UTR) and the 5' UTR of their respective mRNAs. Cellular iron levels affect binding of IRPs to IREs and consequently expression of TfR and ferritin. Moreover, NO(.), a redox species of nitric oxide that interacts primarily with iron, can activate IRP1 RNA-binding activity resulting in an increase in TfR mRNA levels and a decrease in ferritin synthesis. We have shown that treatment of RAW 264.7 cells (a murine macrophage cell line) with NO(+) (nitrosonium ion, which causes S-nitrosylation of thiol groups) resulted in a rapid decrease in RNA-binding of IRP2, followed by IRP2 degradation, and these changes were associated with a decrease in TfR mRNA levels and a dramatic increase in ferritin synthesis. Moreover, we demonstrated that stimulation of RAW 264.7 cells with lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma) increased IRP1 binding activity, whereas RNA-binding of IRP2 decreased and was followed by a degradation of this protein. Furthermore, the decrease of IRP2 binding/protein levels was associated with a decrease in TfR mRNA levels and an increase in ferritin synthesis in LPS/IFN-gamma-treated cells, and these changes were prevented by inhibitors of inducible nitric oxide synthase. These results suggest that NO(+)-mediated degradation of IRP2 plays a major role in iron metabolism during inflammation. PMID- 12547231 TI - Hemochromatosis--neonatal and young subjects. AB - Juvenile hemochromatosis (JH) is an autosomal recessive disease causing iron overload before age 30 in both sexes. JH is characterised by hypogonadism, growth retardation and cardiomyopathy. Linkage of JH to chromosome lq is established in pedigrees throughout Europe. Studies of 29 patients in 20 families of diverse ethnic origin confirm early-onset iron overload. Neonatal hemochromatosis (NH) is a syndrome of unknown origin characterized by congenital cirrhosis or fulminant hepatitis with hepatic and extra-hepatic iron deposits. We assessed 40 infants from 27 families and identified 3 patterns of disease transmission. In 12 of the 27 there was >1 affected infant and in 5 families all infants were affected by NH. In 19 families unaffected children were also born. In 4 families there was bacterial or viral maternal infection associated with NH. In two families, antibodies to DNA or ribonuclear proteins were identified. In 12 families, unaffected children were born to the same parents in the absence of maternal antibodies or infection and without indications of maternal transmission. Consanguinity was observed in 1 family with 4 affected offspring (1 stillbirth + 3 neonatal deaths). Sequence analysis of HFE, beta2M, and both human heme oxygenase genes failed to identify any causal mutations in nuclear NH families but our study points to the existence of a cohort of patients likely to suffer from an autosomal recessive trait. A genome wide scanning study is underway to identify the putative locus. PMID- 12547232 TI - Increased lipid peroxidation and mitochondrial dysfunction in aceruloplasminemia brains. AB - Aceruloplasminemia is characterized by iron accumulation in the brain as well as in visceral organs, due to the absence of ceruloplasmin ferroxidase activity. The neurological symptoms, which include involuntary movements, ataxia, and dementia, reflect the sites of iron deposition. The unique involvement of the central nervous system distinguishes aceruloplasminemia from other inherited and acquired iron storage disorders. Excess iron functions as a potent catalyst of biologic oxidation. An increased iron concentration was associated with increased lipid peroxidation in the brains of three aceruloplasminemia patients. Positron emission tomography showed brain glucose and oxygen hypometabolism. Enzyme activities in the mitochondrial respiratory chain of the basal ganglia were reduced to about 50 and 43%, respectively, for complexes I and IV. Those of the cerebral and cerebellar cortices also were decreased approximately 62 and 65%. These findings suggest that iron-mediated free radicals may contribute to neuronal cell damage through increased lipid peroxidation and the impairment of mitochondrial energy metabolism in aceruloplasminemia brains. PMID- 12547233 TI - Dominant hemochromatosis due to N144H mutation of SLC11A3: clinical and biological characteristics. AB - Hereditary hemochromatosis is classically inherited as a recessive trait but is genetically heterogeneous. Mutations in the HFE and the TFR2 genes account for about 80% of patients and a third locus on chromosome 1q is responsible for juvenile hemochromatosis. We describe here the clinical and biological characteristics of autosomal dominant form of iron overload due to the N144H mutation of the SLC11A3 gene. Clinical signs of iron overload in patients include joint pains, cardiomyopathies, liver fibrosis and hormonal disorders including diabetes mellitus. The main and most common clinical symptoms in this family were joint complaints and early signs of arthrosis. Serum ferritin levels in iron overloaded subjects varied from 31 to 2179 ng/ml and the transferrin saturation from 13 to 88.6%. The iron overload is moderate compared to patients with type 1 hemochromatosis but the deferoxamine test was normal in all patients. The disease in this family segregated as a dominant trait. None of the patients was homozygous or compound heterozygous for any known mutation in the HFE or TFR2 genes. The disease in this family represents a non-classical form of iron overload caused by the N144H mutation in the SLC11A3 gene. The reports of other distinct mutations in SLC11A3 suggest that this gene may be of interest for further etiologic research. PMID- 12547234 TI - The GRACILE syndrome, a neonatal lethal metabolic disorder with iron overload. AB - GRACILE syndrome (Fellman syndrome, MIM 603358), an autosomal recessive metabolic disorder of the Finnish disease heritage, has been diagnosed in 25 infants of 18 families. The incidence is at least 1/47,000 in Finland. The main findings are fetal growth retardation, Fanconi type aminoaciduria, cholestasis, iron overload (liver hemosiderosis, hyperferritinemia, hypotransferrinemia, increased transferrin iron saturation, and free plasma iron), profound lactic acidosis, and early death. The pathophysiology of the metabolic disturbance is unsolved. No significant deficiency of complex III activity of respiratory chain has been found, although we recently showed that the underlying genetic cause is a missense mutation (S78G) in the BCS1L gene and other mutations in that gene have been associated with complex III deficiency. BCS1L encodes a mitochondrial protein, acting as a chaperone in the assembly of complex III. Iron accumulation in liver, a typical feature being less abundant with increasing age, might be a primary abnormality or a secondary phenomenon due to liver dysfunction. In order to decrease the iron overload, three infants have been repeatedly treated with apotransferrin followed by exchange transfusion. Improvement in iron biochemistry occurred, but no clear beneficial effect on the clinical condition was found. Further studies will elucidate the role of iron in the pathophysiology of the disease. PMID- 12547235 TI - Non-invasive liver iron quantification by SQUID-biosusceptometry and serum ferritin iron as new diagnostic parameters in hereditary hemochromatosis. AB - In the HFE-gene era, precise diagnostic parameters remain important to characterize individual iron stores, because the indication for therapy and prognosis are mainly related to the extent of iron loading. The frequently used serum ferritin interferes with non-iron related factors such as inflammation and may produce falsely positive values. We used a SQUID-biosusceptometer in a large series of patients (n = 679) to measure liver iron concentration in the differential diagnosis and therapy control of hereditary hemochromatosis (SQUID = superconducting quantum interference device). This truly non-invasive technique is sensitive, reliable, fast (online results), and also cost-effective when compared to invasive liver biopsy. Recently, ferritin iron content was propagated as a better parameter than ferritin protein. However, we found a poor correlation between ferritin iron and individual liver iron concentrations in patients with iron overload. Ferritin iron saturation varied in a range between 3 and 10%, independent from liver iron concentration. No differences were found between patients with hemochromatosis and secondary iron overload disease. Only patients with liver cell damage had increased ferritin iron saturations. In conclusion the diagnostic values of serum ferritin protein and iron to assess iron overload are limited. PMID- 12547236 TI - Patch-clamp capacitance measurements: new insights into the endocytic uptake of transferrin. AB - Since its introduction by Neher and Sakmann in 1976 the patch-clamp technique has been extensively used to study processes such as signalling and synaptic transmission, but also for monitoring endo- and exocytosis. Since biological membranes behave like electrical capacitors high-resolution measurements of membrane capacitance allow detection of small changes in membrane surface area that accompany exocytosis and endocytosis. We here describe our recent work on patch-clamp capacitance measurements in stably transfected HeLa cells expressing HFE, the hereditary hemochromatosis gene product, under the control of a tetracycline-sensitive promotor. By means of whole-cell and cell-attached techniques we were able to reveal transferrin-induced decreases in membrane capacitance reflecting increased endocytosis at the single cell level. Moreover, cell-attached recordings revealed significant alterations in the formation of single endocytic vesicles. Time-resolved measurements of cell membrane capacitance provide a new methodological approach to study the endocytic uptake of transferrin and its regulation by HFE, the hereditary hemochromatosis protein. PMID- 12547237 TI - Hemochromatosis due to mutations in transferrin receptor 2. AB - A rare recessive disorder which leads to iron overload and severe clinical complications similar to those reported in HFE-related hemochromatosis has been delineated and sometimes called hemochromatosis type 3. The gene responsible is Transferrin Receptor 2 (TFR2), which maps to chromosome 7q22. The TFR2 gene presents a significative homology to transferrin receptor (TFRC) gene, encodes for a transmembrane protein with a large extracellular domain, is able to bind transferrin, even if with lower affinity than TFRC. The TFR2 function is still unclear. The transcript does not contain IRE elements and is not modified by the cellular iron status. At variance with TFRC, interactions between TFR2 and HFE do not occur, at least in their soluble forms. TFR2 is spliced in two alternative forms, alfa and beta. The alfa form is strongly expressed in the liver. The beta form, codified from a start site in exon 4 of the alpha, has a low and ubiquitous expression. Using anti-TFR2 monoclonal antibodies we have confirmed expression of the protein in the liver but also in duodenal epithelial cells, and studied the protein functional behaviour in cell lines, in response to iron addition, iron deprivation and olo-transferrin exposure. Our results suggest a regulatory role of TFR2 in iron metabolism. Five TFR2 homozygous mutations have been documented in HFE3 patients: a nonsense mutation (Y250X); a C insertion that causes a frameshift and a premature stop codon (E60X); a missense mutation (M172K); a 12 basepair deletion in exon 16, that causes 4 aminoacid loss (AVAQ 594-597del) in the extracellular domain of TFR2; a missense mutation in exon 17 (Q690P). The mutation analysis supports the hypothesis that all are private mutations. The pathogenetic role of TFR2 in hemochromatosis has been recently further demonstrated through the targeted expression of the Y250X human mutation in mice, which develop sings of iron overload identical to the human disease. Although the rarity of TFR2 mutations limits their usefulness in diagnostic/screening programs, their study can contribute to a better understanding of the protein function. PMID- 12547238 TI - Seeking candidate mutations that affect iron homeostasis. AB - Hereditary hemochromatosis is characterized by marked variation of expression of the defect: very few homozygotes with the C282Y/C282Y HFE genotype have full blown clinical disease, a larger number show biochemical stigmata of iron overload, and some seem normal biochemically. The following candidate genes have been examined in detail to determine whether polymorphisms in them may be responsible for this variation: transferrin, transferrin receptor 1, transferrin receptor 2, ferritin-L, ferritin-H, IRP1, IRP2, HFE, beta(2) microglobulin, mobilferrin/calreticulin, ceruloplasmin, ferroportin, NRAMP1, NRAMP2 (DMT1), haptoglobin, heme oxygenase-1, heme oxygenase-2, hepcidin, USF2, ZIRTL, duodenal cytochrome b ferric reductase (DCYTB), TNFalpha, keratin 8, and keratin 18. The coding sequence, exon-intron junctions, and promoters of each of these genes was sequenced in DNA from 20 subjects: 5 HFE C282Y/C282Y with clinical disease, 5 HFE C282Y/C282Y with normal/low ferritin levels and no disease, 5 wt/wt with high ferritin and transferrin saturation, and 5 wt/wt normal controls. When coding or promoter polymorphisms were encountered, DNA from large numbers of ethnically defined subjects was examined for these polymorphisms and a relationship between their existence and abnormalities of iron homeostasis was sought. Only in the case of one transferrin mutation did we find a strong relationship between the polymorphism and iron deficiency anemia. The putative genes that affect the expression of HFE mutations remain elusive. PMID- 12547239 TI - Mechanisms of iron mediated regulation of the duodenal iron transporters divalent metal transporter 1 and ferroportin 1. AB - Intestinal iron absorption is regulated by the body's demands for iron. Identification of divalent metal transporter 1 (DMT1) and ferroportin 1 (FPN1) has improved our understanding of iron transport across the intestinal epithelium. Although DMT1 and FPN1 mRNA bear an iron responsive element (IRE) within its untranslated regions which should cause susceptibility to iron mediated posttranscriptional regulation the latter has not been shown so far. The effects of iron perturbations on DMT1 and FPN1 expression were investigated in CaCo2 cells and in primary tissue cultures of human duodenal biopsies by means of Northern Blot, Western Blot, RNA-bandshift and Nuclear Run off analysis. Both DMT1 and FPN1 mRNA levels were increased upon treatment of CaCo2 cells with desferrioxamine, whereas iron treatment resulted in the opposite effect. These changes were paralleled by the respective alterations in DMT1 and FPN1 protein expression. Although desferrioxamine treatment increased the binding affinity of iron regulatory protein-1 to DMT1- and FPN1-IRE, the mRNA half life of DMT1 mRNA remained unchanged. Nuclear run-off analysis then demonstrated that the effects of iron and desferrioxamine on DMT1 and FPN1 mRNA expression are rather due to modulation of transcription of these genes. Our results demonstrate that iron unidirectionally regulates the expression of the two ferrous ion transporters DMT1 and FPN1 by affecting their transcription. This provides evidence for a negative feed-back loop between intracellular iron availability and transmembrane iron transport. PMID- 12547240 TI - Transferrin receptor 1 (TfR1) and putative stimulator of Fe transport (SFT) expression in iron deficiency and overload: an overview. AB - Transferrin Receptor 1 (TfR1) and putative Stimulator of Fe Transport (SFT) represent two different proteins involved in iron metabolism in mammalian cells. The expression of TfR1 in the duodenum of subjects with normal body iron stores has been mainly localized in the basolateral portion of the cytoplasm of crypt cells, supporting the idea that this molecule may be involved in the sensing of body iron stores. In iron deficiency anemia TfR1 expression demonstrated an inverse relationship with body iron stores as assessed by immunohistochemistry with anti-TfR1 antibodies. In iron overload, TfR1 expression in the duodenum differed according to the presence or absence of the C282Y mutation in the HFE gene, being increased in HFE-related hemochromatosis and similar to controls in non-HFE-related iron overload. SFT is characterized by its ability to increase iron transport both through the transferrin dependent and independent uptake, and could thus affect iron absorption in the intestine. Immunohistochemistry using anti-SFT antibodies which recognize a putative stimulator of Fe transport of approximately 80 KDa revealed a localization of this protein in the apical part of the cytoplasm of enterocytes localized at the tip of the villi. The expression of the protein recognized by these antibodies was increased in iron deficiency, as well as in patients carrying the C282Y HFE mutation. Thus, the increased expression of both proteins only in patients with HFE-related hemochromatosis suggests that other factors should be involved in determining non-HFE-related iron overload. PMID- 12547241 TI - Iron deficiency and anaemia in pregnancy: modern aspects of diagnosis and therapy. AB - The prevalence of iron-deficiency anemia in different regions of the world ranges from 12 to 43%. The increased iron requirement in pregnancy and the puerperium carry with it an increased susceptibility to iron deficiency and iron-deficiency anemia and perioperative or peripartal blood transfusion. Prevention and correction presuppose reliable laboratory parameters and a thorough understanding of the mechanisms of iron therapy. The Hb level alone is insufficient to guide management. A complete work-up (ferritin, transferrin saturation) is essential, preferably with hematological indices such as hypochromic and microcytic red cells and reticulocytes, classified by degree of maturity, in particular, before parenteral therapy is given. Since ferritin acts as both an iron-storage and acute-phase protein, it cannot be used to evaluate iron status in the presence of inflammation. A high ferritin level thus requires the presence of an inflammatory process to be eliminated before it can be taken at face value. If the C-reactive protein level is also raised, the soluble TfR concentration can be used, since it is unaffected by inflammation. Inadequate understanding of the complex chemistry of parenteral iron administration was previously responsible for serious side effects, such as toxic and allergic reactions, and even anaphylactic shock, in particular with dextran preparations. However, the current type II iron complexes that release iron to the endogenous iron-binding proteins with a half-life of about 6 hours are not only effective but carry a minimal risk of allergic accident and overload, especially after a comprehensive pretreatment work-up. Our departmental data collected over 8 years and backed by postmarketing experience in 25 countries indicate that iron sucrose complex therapy is a valid first-line option for the safe and rapid reversal of iron-deficiency anemia. PMID- 12547246 TI - Neuroferritinopathy: a window on the role of iron in neurodegeneration. AB - Neuroferritinopathy is a recently recognised genetic disease resulting in a dominantly inherited movement disorder. The condition was mapped by linkage analysis to chromosome 19q13.3 and found to be due to a single adenine insertion in the ferritin light chain (FTL) gene at position 460-461 which is predicted to alter the C terminus of the FTL polypeptide. Clinical features of neuroferritinopathy are highly variable, with chorea, dystonia, and Parkinsonian features predominating in different affected individuals. The most consistent feature is a dystonic dysarthria. Symptoms and abnormal physical signs appear to be restricted to the nervous system and onset is typically in the fourth to sixth decades. Low serum ferritin also characterises this condition. Brain MR imaging of affected patients demonstrates iron deposition in the basal ganglia, progressing over years to cystic degeneration, and brain histochemistry shows abnormal aggregates of ferritin and iron. Now that the molecular basis of the condition is known, therapeutic interventions to reduce or reverse brain iron deposition are being evaluated. This rare disease provides evidence of a central role for iron metabolism in neurodegenerative disorders. PMID- 12547247 TI - Pathogenesis of hyperferritinemia cataract syndrome. AB - Hereditary hyperferritinemia-cataract syndrome (HHCS) is an autosomal dominant disorder characterized by bilateral cataracts and increased serum L-ferritin, in the absence of iron overload. Under physiological conditions, ferritin synthesis is finely regulated at the translational level by iron availability. This regulation is achieved by the high-affinity interaction between cytoplasmic mRNA binding proteins (iron regulatory proteins, IRPs), and mRNA stem-loop structures, known as iron responsive elements (IREs), located in the untranslated regions (UTRs) of the mRNAs. A single IRE is located on the 5' UTR of a series of genes involved in iron metabolism, like L-ferritin, and the binding IRE-IRPs represses these genes translation. The deregulation of ferritin production responsible of HHCS is caused by heterogeneous mutations in the iron regulatory element (IRE) of L-ferritin that interfere with the binding of iron regulatory proteins, disrupting the negative control of L-ferritin synthesis and causing the constitutive up-regulation of ferritin L-chains. The HHCS families originate from different countries of Europe and North America, suggesting that HHCS may be distributed widely throughout the world and not sporadic, whereas its prevalence remains to be established. The lens seems to be particularly sensitive to the increased amount of L-ferritin and the alteration of the proteic equilibrium in this tissue can be responsible of the cataract. In spite of the elucidation of the genetic basis, the genotype phenotype correlation is not clear. Recently, a study based on the thermo-denaturation profile and dissociation constant of the IRE-IRP complex performed for several mutated IREs has provided evidence for a possible correlation between heterogeneous IRE mutations and serum ferritin levels. On the other hand, the in vivo relevance of these conclusions has not been determined completely. A clinical variability among subjects sharing the same mutation, whether they belonged to the same family or not, has also been demonstrated. These findings suggest that, besides the L-ferritin IRE genotype, additional factors are likely to modulate the lens involvement and the rate of progression to severe cataract in HHCS patients. PMID- 12547248 TI - Iron metabolism and mitochondrial abnormalities in Friedreich ataxia. AB - Friedreich ataxia is an autosomal recessive disease causing degeneration in the central and peripheral nervous system, cardiomyopathy, skeletal abnormalities and increased risk of diabetes. It is caused by deficiency of frataxin, a highly conserved nuclear-encoded mitochondrial protein. The genetic mutation found in 98% of Friedreich ataxia chromosomes is the unstable hyperexpansion of a GAA triplet repeat in the first intron of the gene. The expanded GAA repeat, by adopting an abnormal triple helical structure, impairs frataxin transcription. Longer repeats cause a more profound frataxin deficiency and are associated with earlier onset and increased severity of the disease. Yeast cells deficient in the frataxin homologue (Deltayfh1) become unable to carry out oxidative phosphorylation, lose mitochondrial DNA, accumulate iron in mitochondria, show unregulated high expression of high affinity iron uptake, and have an increased sensitivity to oxidative stress. Loss of respiratory competence in Deltayfh1 is iron-dependent. Additional properties of these cells include a deficiency of iron sulfur cluster containing proteins (ISPs) and impaired iron efflux out of mitochondria. Evidence of oxidative stress, mitochondrial dysfunction, deficiency of multiple ISPs and iron deposits are also found in the human disease and in mouse models. The primary function of frataxin is still unknown, however much recent evidence suggests that it enhances iron-sulfur cluster synthesis and protects iron from free radical-generating reactions. The search for frataxin function stimulated more investigations on the role of mitochondria in cellular iron homeostasis. Their results suggest that these organelles may play a central role in controlling iron homeostasis, which is not surprising considering that they are the major cellular site where this metal is utilized. I propose a model, valid in yeast as well as in higher eukaryotes, in which iron transport into mitochondria is directly coupled to its uptake at the cell membrane and iron transport out of mitochondria depends on adequate iron-sulfur cluster synthesis. Regulatory mechanisms in the cytosol would then sense a post-mitochondrial iron pool. Much circumstantial evidence from genetically manipulated yeast and from human diseases supports this model. PMID- 12547252 TI - Role of Ets-1 in erythroid differentiation. AB - Members of the Ets gene family are known to be expressed in the hematopoietic tissue and some of them play a pivotal role in normal hematopoietic cell development. Ets-1 gene expression was analyzed in Friend Leukemia Cells (FLC) induced to erythroid differentiation by DMSO. We show that the level of Ets-1 protein and its binding activity decreases in FLC along erythroid differentiation of primary human progenitors. The same behavior was observed during normal erythroid differentiation. Moreover, FLC constitutively expressing Ets-1 show a decrease in TfR gene expression, globin mRNA and hemoglobin synthesis. These data indicate that a decrease in Ets-1 binding activity is required for a normal erythroid maturation and that a deregulated expression of this transcription factor may interfere with terminal erythroid differentiation. PMID- 12547253 TI - Natural freeze-tolerance in hatchling painted turtles? AB - Hatchlings of the North American painted turtle (Family Emydidae: Chrysemys picta) typically spend their first winter of life inside a shallow, subterranean hibernaculum (the natal nest) where life-threatening conditions of ice and cold commonly occur. Although a popular opinion holds that neonates exploit a tolerance for freezing to survive the rigors of winter, hatchlings are more likely to withstand exposure to ice and cold by avoiding freezing altogether-and to do so without the benefit of an antifreeze. In the interval between hatching by turtles in late summer and the onset of wintery weather in November or December, the integument of the animals becomes highly resistant to the penetration of ice into body compartments from surrounding soil, and the turtles also purge their bodies of catalysts for the formation of ice. These two adjustments, taken together, enable the animals to supercool to temperatures below those that they routinely experience in nature. However, cardiac function in hatchlings is diminished at subzero temperatures, thereby compromising the delivery of oxygen to peripheral tissues and eliciting an increase in reliance by those tissues on anaerobic metabolism for the provision of ATP. The resulting increase in production of lactic acid may disrupt acid/base balance and lead to death even in animals that remain unfrozen. Although an ability to undergo supercooling may be key to survival by overwintering turtles in northerly populations, a similar capacity to resist inoculation and undergo supercooling characterizes animals from a population near the southern limit of distribution, where winters are relatively benign. Thus, the suite of characters enabling hatchlings to withstand exposure to ice and cold may have been acquired prior to the northward dispersal of the species at the end of the Pleistocene, and the characters may not have originated as adaptations specifically to the challenges of winter. PMID- 12547254 TI - Long-term in vitro maintenance of neuromuscular junction activity of Drosophila larvae. AB - The larval Drosophila neuromuscular junction (NMJ) has proven to be an excellent system to test fundamental aspects of synaptic transmission, such as relationships among ion channel function, subtypes of glutamate receptors, and the functions of synaptic proteins in the presynaptic compartment. Recent advances in understanding bi-directional communication between nerves and muscles of Drosophila are helping uncover developmental as well as maintenance cues that could be applicable to all chemical synapses. The development of HL3 medium makes it possible to record synaptic responses at NMJs for prolonged periods of time. We demonstrate that media commonly used to culture CNS neurons and imaginal disks of Drosophila such as Schneider's and M3 completely block glutamatergic synaptic transmission at the NMJ. The depressed postsynaptic excitatory junction potentials (EJPs) partially recover from exposure to such media shortly after switching to the HL3 medium. Preliminary results from NMJs of filleted 3rd instar larvae for 4 days in vitro bathed in a modified HL3 medium show great promise. The resting membrane potential and the EJP amplitudes after 4 days in vitro are normal. These results demonstrate the possibility for chronic studies of developmental regulation in culture, which in some cases are impractical in the whole animal. PMID- 12547255 TI - Respiratory responses of the mysid Gastrosaccus brevifissura (Peracarida: Mysidacea), in relation to body size, temperature and salinity. AB - The mysid Gastrosaccus brevifissura (Peracarida: Mysidacea) is widely distributed in southern Africa and is thought to be important in the functioning of estuarine systems. This mysid may experience highly variable physicochemical conditions, and its physiological responses to these are of interest considering its ecological role. This study presents data on the metabolic physiology in relation to body length, temperature (15-30 degrees C) and salinity (15-35 psu) of a G. brevifissura population on the sub-tropical eastern seaboard of South Africa. Oxygen consumption rate was linearly related to size (for body lengths ranging from 3 to 10 mm) and varied among individuals from 0.67 to 6.51 microgram h(-1), dependent on environmental conditions. Oxygen consumption rate was largely independent of salinity variation between 20 and 35 psu, although was significantly depressed at 15 psu. Aerobic rate generally increased with an acute increase in temperature (Q(10)=2.147), but was not affected by 7 days of acclimation at either 15 or 25 degrees C. The lack of a metabolic adjustment to meet the additional energetic demands associated with a decline in salinity may well be a factor limiting the estuarine distribution of G. brevifissura. Even though feeding behaviour substantially changes between summer and winter, this may best be explained by food availability or other ecological factors, rather than a metabolic adjustment, considering the apparent lack of metabolic acclimation. PMID- 12547256 TI - Cyclic AMP level and phosphodiesterase activity during 17alpha,20beta-dihydroxy-4 pregnen-3-one induction and theophylline inhibition of oocyte maturation in the catfish, Clarias batrachus. AB - This study directly tested the hypothesis that the induction of oocyte maturation in the catfish Clarias batrachus is followed by a transient decrease in oocyte cyclic AMP (cAMP) level that is due to an increase in phosphodiesterase (PDE) activity. Further, the PDE inhibitor theophylline was used to investigate the possible role of PDE in the maturation-inducing action of 17alpha,20beta dihydroxy-4-pregnen-3-one (17alpha,20beta-DP), the physiological maturation inducing steroid of this catfish species. The results obtained from batches of oocytes taken from the same donor at the same time clearly show a close relationship between dose-dependent induction of germinal vesicle breakdown (GVBD) and PDE activity with a concomitant decrease in cAMP in the oocytes treated with different concentrations of 17alpha,20beta-DP. In contrast, theophylline prevents GVBD and inhibits PDE activity by promoting cAMP accumulation in oocytes. A time-dependent decrease in PDE activity and an increase in cAMP content with a marked inhibition of GVBD were recorded even in oocytes pre-stimulated with 1 microgram/ml 17alpha,20beta-DP for 6 h and then treated with 1 mM theophylline for various times. These results suggest that cAMP plays a key role in the regulation of oocyte maturation in C. batrachus which may be mediated by PDE activity. PMID- 12547257 TI - A sea anemone's environment affects discharge of its isolated nematocysts. AB - Nematocysts were isolated from individuals of Calliactis tricolor maintained under different feeding schedules or in different salinities in an attempt to determine how these culture conditions influence the discharge of isolated nematocysts. In addition, the discharge frequencies of nematocysts isolated from two different populations of sea anemones found in two different environments were also compared. Undischarged acontial nematocysts were isolated by extrusion into 1 M sodium citrate and were then treated with 5 mM EGTA to initiate discharge. Nematocysts isolated from anemones maintained under three different feeding schedules showed significantly different responses to the test solution. Nematocysts isolated from anemones maintained in two different salinities did not differ significantly in discharge frequency. Nematocysts isolated from individuals from two separate populations of C. tricolor responded significantly differently to 5 mM EGTA and to deionized water, and these responses also depended upon the isolation solution used. Environmental conditions are known to have an impact on the physiological state of most organisms, but this is the first study providing evidence that the environment or feeding state of an anemone affects discharge of isolated nematocysts. Inherent differences in ionic and osmotic characteristics among nematocysts could explain some of the ambiguities when comparing past studies of isolated nematocyst discharge. PMID- 12547258 TI - Dynamic energy and time budgets in hummingbirds: a study in Sephanoides sephaniodes. AB - Temperature and diet quality are two of the most important factors affecting the dynamic regulation of animal energy budgets. Because hummingbirds are very sensitive to energy stress, we used Green-backed Firecrowns (Sephanoides sephaniodes) to test the dynamics of their energy budget under different energetic challenges in chronic conditions (20 days). Experimental groups were: HQ-TNZ (high quality diet and thermoneutrality), HQ-LT (high quality diet and low temperature), LQ-TNZ (low quality diet and thermoneutrality), and LQ-LT (low quality diet and low temperature). Analysis of behavioral, morphological, and physiological variables revealed that thermal and dietary factors affect time and energy budgets independently. Hummingbirds increased energy intake during the first day of acclimation at LT, but after second day, the LQ-LT group did not maintain their energy intake and began to loose body mass. Moreover, diet quality affected digestive organs. The intestine, gizzard, liver and kidneys all increased in size when food quality was lowest, probably to obtain more food per feeding event and to more efficiently process the ingested food. Exposure to low ambient temperatures affected the most metabolically productive organs such as the heart, lungs, and muscular mass. Lower temperature increased basal and daily energy expenditure, and changed the time budget. Sephanoides sephaniodes spent more time perching when their energy balance was close to be negative. We suggest that energy budget regulation in hummingbirds does not reside exclusively in the energy output nor in the energy-input but in both pathways. PMID- 12547259 TI - Calcium, phosphorus and adenylate levels and Na(+)-K(+)-ATPase activities of prawn, Macrobrachium nipponense, during the moult cycle. AB - Changes in calcium and phosphorus concentrations, adenylate (AMP, ADP and ATP) levels, and ratios and ATPase activities of Macrobrachium nipponense were investigated during the moult cycle. Ca level in the exoskeleton was lowest in early postmoult (stage A), increasing at stages B and through intermoult (stage C) and peaking in premoult (stage D1 and D2). The P concentrations in the exoskeleton and muscle in late premoult and early postmoult stages were higher than those at other moult stages, and were lowest in the intermoult. Muscle adenylate energy charge (AEC) changed with moult stages, and was in agreement with the change in inorganic P level in the muscle. AEC may be a direct indicator of energy metabolic activity during the moult cycle. ATP/ADP and ATP/AMP ratios in premoult and postmoult stages were higher than that in intermoult stage. Na(+) K(+)-ATPase activities of gills, muscles and hepatopancreatic of prawns were higher in early postmoult and late premoult animals, whereas they were lower in late postmoult, intermoult and early premoult animals. Gill residual ATPase activity was significantly higher in postmoult animals, while the peak value of hepatopancreatic residual ATPase activity appeared in intermoult stage. PMID- 12547260 TI - GC-MS and NMR analysis of Phyllidiella pustulosa and one of its dietary sources, the sponge Phakellia carduus. AB - Gas-chromatographic and mass-spectrometric and NMR analyses of the lipophilic extract of the nudibranch Phyllidiella pustulosa and the sponge Phakellia carduus clearly showed that the sponge formed a major part of the nudibranchs diet. The analyses also indicated that the nudibranch accumulates some of the sponge metabolites in preference to others. The main components of the extracts were identified as the new natural product 10-isothiocyano-4-cadinene (1), axisonitrile-3 (2), and a number of other sesquiterpenes similar to 1 and 2. Also positively identified, in the extract of the sponge were the two sterols (3beta,22E)-ergosta-5,8,22-trien-3-ol (4) and (3beta,22E)-stigmasta-5,7,22-trien 3-ol (5), and in the nudibranch extract caryophyllene (3). The new natural product 10-isothiocyano-4-cadinene (1) was shown to have moderate antiplasmodial activity (IC(50) 1.5 microgram/ml towards Plasmodium falciparum clones K1, and NF54). PMID- 12547261 TI - Identification and localization of eight distinct hormone-producing cell types in the pituitary of male Atlantic halibut (Hippoglossus hippoglossus L.). AB - The eight distinct hormone-producing cell types in the adenohypophysis of male Atlantic halibut (Hippoglossus hippoglossus L.) were identified and localized using immunohistochemistry and in situ hybridization. Lactotropes either occupied most of the rostral pars distalis (RPD) or they were arranged in follicular structures located along the periphery of the RPD. Corticotropes were confined to a thin layer of RPD cells bordering the pars nervosa (PN). The somatotropes were arranged in multicellular layers bordering the highly convoluted PN penetrating the proximal pars distalis (PPD), while thyrotropes, scattered in small islets in between the somatotropes, were located in the centro-dorsal part of the PPD. Gonadotropes were found throughout the PPD. Immunoreactivity to glycoprotein alpha and luteinizing hormone beta-subunit was also observed along the periphery of the pars intermedia (PI), indicating that a thin extension of the PPD surrounded the PI. In situ hybridization showed that follicle-stimulating hormone and luteinizing hormone were produced in distinct cells of the PPD. PI contained somatolactotropes bordering the highly convoluted PN, and melanotropes that showed positive immunostaining against both anti-alpha-melanocyte-stimulating hormone and anti-beta-endorphin. The general cellular organization was similar to that of other teleost fish. These results lay the basis for future investigations on Atlantic halibut pituitary physiology. PMID- 12547262 TI - Circulating leptin response to feeding and exogenous infusion of insulin in sheep exposed to thermoneutral and cold environments. AB - Leptin has been shown to regulate feed intake and energy expenditure. Insulin stimulates leptin secretion in rodents, but its action on leptin secretion is still obscure in ruminants. If insulin stimulates leptin secretion in ruminants, circulating leptin concentrations may change during exposure to cold, because of fluctuating insulin secretion and action in the cold environment. The present experiment was designed to determine whether feeding or exogenous administration of insulin affects circulating leptin levels in sheep exposed to thermoneutral and cold environments. Suffolk rams that were shorn and fed a diet once daily were subjected to a thermoneutral (20 degrees C) or cold (0 degrees C) environment for at least 1 week. Overall mean concentrations of plasma leptin in the feeding experiment were lower (P<0.05) in the cold environment than in the thermoneutral environment. Plasma leptin levels remained relatively unchanged after feeding in both environments, though plasma insulin response to feeding in both environments increased (P<0.01). The euglycemic clamps (insulin infusion rate: 4 mUkgBW(-1)min(-1) for 2 h) increased (P<0.01) circulating leptin concentrations in the thermoneutral, but not in the cold environment. These results suggest that lower circulating leptin levels in ruminants exposed to the cold environment could be partly due to the depressed insulin action on leptin secretion. PMID- 12547263 TI - Effect of partial substitution of dietary protein by a single gluconeogenic dispensable amino acid on hepatic glucose metabolism in rainbow trout (Oncorhynchus mykiss). AB - Our objective was to understand the influence of dietary gluconeogenic amino acids on hepatic glucose metabolism in rainbow trout (Oncorhynchus mykiss). We analyzed the effects of partial substitution of dietary protein by a single gluconeogenic dispensable amino acid (DAA: alanine, aspartic acid or glutamic acid), on the regulation of hepatic glycolytic and gluconeogenic enzymes. We fed juvenile rainbow trout with isonitrogenous and isoenergetic diets in which part of nitrogen from fishmeal was replaced by nitrogen from one of the three DAA. Fish were fed over 9 weeks and samples withdrawn 6 h after feeding or 5 days after food deprivation. Our data did not show a clear effect of an excess of DAA on activities of glycolytic enzymes (glucokinase and pyruvate kinase) compared to the control diet. In contrast, feeding caused a significant repression of gluconeogenic enzyme activities (glucose-6-phosphatase, fructose-1,6 bisphosphatase and mitochondrial phosphoenolpyruvate carboxykinase) only in fish fed the three DAA substituted diets. However, these differences were insufficient to affect postprandial glycemia significantly. In conclusion, an excess of dietary DAA tested does not seem to modify glycemia or to have a negative impact on dietary carbohydrate utilization in rainbow trout. PMID- 12547264 TI - Expression and localization of carbonic anhydrase in bovine mammary gland and secretion in milk. AB - Little attention has been paid to carbonic anhydrase VI (CA VI), a secretory type isozyme, in the bovine mammary gland, although the gland is an important exocrine gland and CA VI is known to localize in exocrine glands such as salivary and lacrimal glands in various animal species. In the present study mRNA expression and protein localization of CA VI in isolated gland tissues and in cloned epithelial cells from the mammary gland of Holstein cows (Bos taurus) were observed by reverse transcript polymerase chain reaction and immunocytochemistry. Also, changes of CA VI concentrations in milk were measured for 2 months postpartum by an enzyme-linked immunosorbent assay. CA VI gene expression was detected in the gland tissues and epithelial cells, and CA VI protein was localized in the cytoplasm of the epithelial cells. Colostrum contained the highest concentration of CA VI protein (100 ng/ml), decreasing in an exponential manner (P<0.001). We conclude that bovine mammary epithelial cells synthesize and secrete CA VI in colostrum at higher concentration than in normal milk, implying its role to compensate for low CA VI secretion in neonatal calves. PMID- 12547265 TI - Hypotonic-induced transport pathways in Xenopus laevis erythrocytes: taurine fluxes. AB - Taurine fluxes in Xenopus laevis red cells were studied in vitro in media of different tonicities. Both influx and efflux increased 3-10 times reversibly when dilution of the medium exceeded 30%. The absolute values of uptake ranged between 5 and 30 micromol/l cells.h at extracellular taurine concentration of 1 mmol/l, but is poorly selective as almost the same uptake was measured for choline and sucrose. Q(10) of 2.77 and an activation energy of 71.90+/-7.37 kJ/mol were calculated for the uptake process. Taurine uptake was reduced 50% in the absence of Cl(-), whereas the alkali cations (Na(+), K(+), Li(+) and Rb(+)) supported it similarly. Taurine uptake was greatly increased in Ca(2+)-free solution, and was inhibited by alkaline pH. The inhibitor of anion exchange protein, 4,4' diisothiocyanatostilbene-2,2'-disulphonic acid (IC(50)=25 microM) and the Cl(-) channel blockers 5-nitro-2-(3-phenylpropylamino) benzoic acid and [(dihydro indenyl) oxy] alkanoic acid (IC(50)<20 microM) inhibited taurine uptake effectively. Isoproterenol did not affect taurine uptake in isotonic, nor in hypotonic solution. The uptake was reduced slowly to near the original, control level within 15-30 min in hypotonic solutions, indicating deactivation of the hypotonic-induced taurine pathway. PMID- 12547266 TI - Blood steroid profile and in vitro steroidogenesis by ovarian follicles and testis fragments of adult sea lamprey, Petromyzon marinus. AB - The main purpose of the study was to identify the principal gonadal steroids synthesized by male and female sea lampreys, Petromyzon marinus. To achieve this, we used high performance liquid chromatography to separate the steroids in the serum of sexually mature animals, and to separate the steroids produced by gonadal tissue incubated in the presence of radiolabelled precursor steroids, as a means of identifying the major steroidogenic pathways. We were unable to detect evidence of the 'classical' steroids, such as 17beta-estradiol (E(2)) or testosterone (T) in the serum of either male or female lampreys. Instead, the principal chromatographic peaks contained very polar compounds that had elution times consistent with 15alpha-hydroxylated estrogens and androgens, and there were sex-specific differences in the chemical nature and the quantity of these compounds. Testis fragments or ovarian follicles co-incubated with tritium labelled pregnenolone ([3H]P(5)), 17-hydroxyprogesterone ([3H]17OHP(4)), or androstenedione ([3H]A(4)), provided additional confirmation that the gonads synthesize a range of very polar steroids, and the metabolites found were consistent with the presence of a 15alpha-hydroxylated (15alphaOH) metabolic pathway common to testis and ovary. For ovarian tissue, the major 'end product' metabolites from all three precursors were 15alphaOH-estrogens, and for testis tissue 15alpha-hydroxyprogesterone (15alphaOHP(4)) and 15alpha hydroxytestosterone (15alphaOHT) and small amounts of 15alphaOH estrogen. Small amounts of E(2) were also produced by both ovarian (all substrates) and testicular tissue (some substrates). Although it was assumed that the E(2) was synthesized via the aromatization of T, [3H]T was not found as an intermediate metabolite. The study suggests that the principal gonadal steroids in sea lamprey are 15alpha-OH compounds, and that only small amounts of E(2) or T are synthesized by the gonads at this stage of reproductive development. There was no direct evidence of progesterone (P(4)) synthesis from [3H]P(5), although the metabolites synthesized by both testis and ovary were indicative of a metabolic pathway that involved P(4) as an intermediate. PMID- 12547267 TI - A complement component C3a-like peptide stimulates chemotaxis by hemocytes from an invertebrate chordate-the tunicate, Pyura stolonifera. AB - Recent evidence suggests that the complement system evolved as a critical host defence mechanism among invertebrates, long before the origin among vertebrates of adaptive immune responses mediated by somatically re-arranging antibodies. The current study supports that contention by identifying a complement component C3a like peptide in the tunicate, Pyura stolonifera. Activation of P. stolonifera serum with common inflammatory elicitors (lipopolysaccharide and zymosan) resulted in the proteolytic generation of an 8.5 kDa peptide, and concomitantly conferred chemoattractant activity on the serum. The 8.5 kDa peptide shares substantial amino acid sequence homology with a previously characterised tunicate complement component C3-like protein (72% amino acid identity in an 18 amino acid overlap). It is also recognised by an anti-C3 antiserum that is known to cross react with tunicate C3 homologues. Hemocyte migration assays performed with the 8.5 kDa peptide that had been partially purified by gel filtration confirmed that the molecule acts as a powerful chemotactic agent. This suggests that the proteolytic activation of tunicate C3-like molecules can initiate inflammatory responses involving cellular recruitment by liberating a pro-inflammatory peptide akin to the vertebrate anaphylatoxin, C3a. PMID- 12547268 TI - Milk composition of free-living yellow-pine chipmunks (Tamias amoenus): temporal variation during lactation. AB - Milk is the sole food source of mammals during early postnatal development, and its composition may be modified to meet changing nutritional and energetic demands from birth to weaning. These demands are especially acute in small mammals that breed in highly seasonal environments. We investigated the temporal course of milk composition during lactation in free-living yellow-pine chipmunks, Tamias amoenus, a small altricial omnivorous rodent that produces a single annual litter immediately after emerging from hibernation. Over the course of lactation the total energy concentration of milk increased more than two-fold, to approximately 13 kJ ml(-1). The main component of the milk was lipids, which increased from 10 to 30% of total milk content by wet mass. Proteins increased from approximately 5 to 10%, whereas carbohydrates remained low, at 4-5%. The progressive augmentation of milk energy content during lactation contributes to the increased demands of rapid growth in this altricial species. PMID- 12547269 TI - Ontogenic development of intestinal disaccharidases in the precocial rodent Octodon degus (Octodontidae). AB - We studied the ontogeny of the intestinal brush border disaccharidases sucrase and lactase in the precocial rodent Octodon degus. Sucrase hydrolyze sugars from plants while lactase hydrolyzes sugars from milk. Enzyme expression varied inversely with dietary changes according to the developmental pattern. All new born pups had high lactase and low sucrase activities. Also, a negative correlation between sucrase and lactase activity was found, supporting the economic design hypothesis for the intestinal tract. Profiles for development of sucrase expression exhibit some differences among precocial species, and in O. degus is correlated with the slower transition from milk to solid food consumption at weaning. PMID- 12547270 TI - Differential chemosensory feeding behaviour by three co-occurring mysids (Crustacea, Mysidacea) from southeastern Tasmania. AB - Three mysid species showed differences in chemosensory feeding as judged from stereotyped food capturing responses to dissolved mixtures of feeding stimulant (either betaine-HCl or glycine) and suppressant (ammonium). The strongest responses were to 50:50 mixtures of both betaine-ammonium and glycine-ammonium solutions. In general, the response curve to the different mixtures tested was bell-shaped. Anisomysis mixta australis only showed the normal curve in response to the glycine-ammonium mixture. The platykurtic curve for Tenagomysis tasmaniae suggests a less optimal response to the betaine-HCl-ammonium solution. Paramesopodopsis rufa reacted more strongly to the betaine-ammonium than to the glycine-ammonium solutions, and more individuals of this species responded to both solutions than the other two species. It is suggested that these contrasting chemosensitivities of the three coexisting mysid species serve as a means of partitioning the feeding niche. PMID- 12547271 TI - Unusual physiology of scale-less carp, Gymnocypris przewalskii, in Lake Qinghai: a high altitude alkaline saline lake. AB - The scale-less carp (Gymnocypris przewalskii) inhabits Lake Qinghai located on the Qinghai-Tibet plateau (elevation, 3200 m) in western China. The lake waters are alkaline (pH 9.4, titratable alkalinity=30 mmol l(-1)), Mg(2+)-rich (18.7 mmol l(-1)), Ca(2+)-poor (0.30 mmol l(-1)) and saline (9 per thousand ). These fish make annual spawning migrations into freshwater rivers. We investigated the physiology of nitrogen excretion and ionoregulation of fish from the lake and river. Fish from both waters were ammonotelic, although ammonia-N excretion rates were lower in lake fish (175 vs. 344 micromol kg(-1) h(-1), P<0.05) resulting in unusually high levels of ammonia in blood plasma (2.23 vs. 0.32 mmol l(-1)), bile, liver, muscle and brain. Exposure to 0.4 mmol l(-1) total ammonia in lake water ([NH(3)]=0.16 mmol l(-1)) killed fish within 8 h. River fish survived exposure to 1.0 mmol l(-1) total ammonia in river water at pH 8.0 ([NH(3)]=0.023 mmol l(-1)) for 24 h suggesting high ammonia tolerance in lake fish. High glutamate dehydrogenase and glutamine synthetase activities in tissues probably allow the fish to alleviate ammonia toxicity by amino acid accumulation. Neither lake nor river fish relied on urea excretion to remove excess N. Urea-N excretion rates were below 20 micromol kg(-1) h(-1) for both groups, and levels of urea in plasma and tissues were moderate. When exposed to elevated ammonia, urea-N excretion increased slightly (approximately 50 micromol kg(-1) h(-1)) and liver and muscle urea levels increased in the river fish. Plasma ion levels were within the range typical of cyprinids, but river fish had significantly higher plasma [Na(+)] and [Cl(-)] and lower [K(+)] than fish from the lake. During 48-h lake-to river water transfer, plasma Na(+) and Cl(-) levels rose significantly. Significantly higher Na(+)/K(+)-ATPase activity in the gills of river fish may be related to the higher plasma ion levels. Plasma [Mg(2+)] and [Ca(2+)] were tightly regulated despite the great differences in the lake and river water levels. PMID- 12547272 TI - Body water handling in response to hypertonic-saline induced diuresis in fasting northern elephant seal pups (Mirounga angustirostris). AB - During natural fasting conditions in postweaned northern elephant seal (NES) (Mirounga angustirostris) pups, urinary water loss is minimized and percent total body water (TBW) is maintained constant. However, following infusion of hypertonic saline, glomerular filtration rate (GFR) and urine output increased in fasting pups. Therefore, we quantified the magnitude of the hypernatremia-induced diuresis relative to the animal's total body water (TBW) pool and the percentage of filtered water reabsorbed. Following a 24 h control period, naturally fasting NES pups (n=7) were infused (4 ml min(-1)) with hypertonic saline (16.7%) at a dose of 3 mmol NaCl kg(-1) body mass. Total body water was estimated prior to infusion by tritium dilution, GFR was estimated by standard creatinine clearance, and urine output (V) was measured for 24 h during the control and post infusion periods. Percentage of filtered water reabsorbed was calculated as (1 (V/GFR))x100. Twenty-four hours following the infusion, GFR (control: 69+/-12 ml min(-1) and post-infusion: 118+/-19 ml min(-1); mean+/-S.E.) increased 77+/-28% above control and the percentage of filtered water reabsorbed was decreased 0.4+/ 0.1%. The increase in urine output (control: 218+/-47 ml d(-1) and post-infusion: 883+/-92 ml d(-1)) accounted for 1.7+/-0.2% of the pups' TBW. The hypernatremia induced diuresis was accompanied by the loss of body water indicating the lack of water retention. Although the 77% increase in GFR was only associated with a 0.4% decrease in the percentage of filtered water reabsorbed, this decrease was significant enough to result in a 4-fold increase in urine output. Despite the observed diuresis, fasting NES pups appear to possess an efficient water recycling mechanism requiring only a small percentage of body water to excrete an excess salt load. This water recycling mechanism may allow pups to avoid negative perturbations in body water as they initiate feeding in a marine environment following the fast. PMID- 12547273 TI - Effects of fasting on growth hormone/insulin-like growth factor I axis in the tilapia, Oreochromis mossambicus. AB - Effects of fasting on the growth hormone (GH)-insulin-like growth factor I (IGF I) axis were examined in the tilapia (Oreochromis mossambicus) acclimated to fresh water. Fasting for 2 weeks resulted in significant reductions in body weight, specific growth rate and hepatosomatic index in both males and females. Significant reductions in specific growth rates were observed after 1 and 2 weeks in both sexes, although the decrease in body weight was not significant in the female. A significant reduction was also seen in the condition factor of females after 2 weeks. No change was seen in the gonadosomatic index in either sex. Two weeks of fasting also produced a significant reduction in plasma IGF-I but not in plasma GH, prolactin (PRL(188)) or cortisol. Significant reductions in the hepatic IGF-I mRNA were seen in both sexes. On the other hand, a significant increase was observed in cortisol receptor mRNA in the female liver. Plasma IGF-I levels were correlated significantly with specific growth rate, condition factor and hepatosomatic index, indicating that plasma IGF-I is a good indicator of growth in the tilapia. No change was seen in plasma glucose or osmolality after 2 weeks of fasting. During fasting, tilapia appears to convert metabolic energy from growth to basal metabolism including maintenance of ion and water balance. PMID- 12547274 TI - Mitochondrial K(ATP) channels and sarcoplasmic reticulum influence cardiac force development under anoxia in the Amazonian armored catfish Liposarcus pardalis. AB - The contribution of alterations in mitochondrial K(ATP) channel activity and the sarcoplasmic reticulum (SR) to anaerobic cardiac function in the anoxia tolerant armored catfish Liposarcus pardalis were assessed. K(ATP) channels contribute to hypoxic cardioprotection in mammals, but little is known of their action in more hypoxia tolerant animals. Anoxia resulted in a decrease in force in isometrically contracting ventricle strips to approximately 40% of the pre-anoxic level. This was maintained for at least 2 h. Upon reoxygenation, hearts recovered to the same level as control preparations. Treatment with 5-hydroxydecanoic acid (5HD), a specific mitochondrial K(ATP) blocker significantly increased force in preparations during anoxia and caused hypercontracture at reoxygenation. Ryanodine, a specific inhibitor of SR function, significantly increased force loss in ventricle preparations under anoxia. Results show that mitochondrial K(ATP) channel activity and SR function are important in anaerobic and post anaerobic contractility in armored catfish heart. PMID- 12547275 TI - Thermal and respiratory control in young rats exposed to cold during postnatal development. AB - We questioned to what extent sustained increases in metabolic rate during the neonatal period may influence the development of thermal and respiratory control. Male rats were exposed to cold (14 degrees C) for the first 3 weeks, which increased metabolic rate with small effects on body growth. Measurements were performed at 1 month of age, when the body weight of the Cold group averaged approximately 88% of Controls. In Cold rats, the concentration of the uncoupling protein of the brown adipose tissue was increased. Acute exposures to different ambient temperatures (5, 15, 25 and 35 degrees C) provoked changes in body temperature similar in Cold and in Control rats. At these temperatures, small differences in the absolute values of oxygen consumption (Vdot;(O(2))) between the two groups could be explained by the differences in body weight. Hematocrit and lung weight of Cold rats were as in Controls, but the lung protein-DNA ratio was increased because of a drop in lung cellularity. The resting ventilation oxygen consumption ratio (Vdot;(E)/Vdot;(O(2))) was similar between Cold and Controls. Also the changes in Vdot;(O(2)) and Vdot;(E) during acute hypoxia (10% O(2)) or hypercapnia (5% CO(2)), and the corresponding hyperventilatory responses (increases in Vdot;(E)/Vdot;(O(2))) did not significantly differ between the two groups. In conclusion, in the rat, the increased metabolic requirements caused by cold exposure during the early postnatal phases improved the thermogenic capacity, while having negligible impact on the development of respiratory control. PMID- 12547276 TI - Bioelectric field potentials of the ventilatory muscles in the crayfish. AB - This study investigated the pattern of the electrical field potentials recorded near the prebranchial chamber of the native crayfish Astacus leptodactylus. Inside the prebranchial chamber, the electrical waves had maximal amplitude and showed 2-4 peaks per cycle. Potentials with the same frequency but smaller amplitude and simpler shape were also recorded outside the chamber, near the edge of the carapace, and at some distance towards the caudal direction. Correlation of these electrical potentials with movements of the scaphognathite in intact Procambarus cubensis with a transparent external wall of the prebranchial chamber has shown a high coincidence both in rate and phase of the two processes. The electrical activity picked up by an electrode located in the prebranchial chamber or near it represents the cumulative electrical field generated by the muscles moving the scaphognathite, and may be termed as the electroscaphognathitegram (ESG). The correlation of the mechanoscaphognatitegram determined optically from the magnified image of the scaphognathite and the ESG allowed us to suggest that the ascending phase of single electrical wave corresponds to the activation of the muscles responsible for downward movement of the scaphognathite, whereas its descending phase reflects its upwards movement. PMID- 12547277 TI - Control of cardiovascular function in the icefish Chionodraco hamatus: involvement of serotonin and nitric oxide. AB - The involvement of nitric oxide (NO) in the branchial circulation and cardiac performance of the Antarctic hemoglobinless icefish Chionodraco hamatus was investigated using isolated and perfused head and working heart preparations. In the branchial vasculature under basal (i.e. unstimulated conditions), the nitric oxide synthase (NOS) inhibitor L-NIO (L-N(5)-(1-iminoethyl) ornithine, 10(-5) and 10(-4) M), caused a marked vasoconstriction (20%), indicating a basal nitrergic vasodilator tone, while the dose-response curve of the NO donor SIN-1 (3 morpholinosydnonimine) showed a dose-dependent vasodilator effect. Acetylcholine induced a dose-dependent branchial vasoconstriction mediated by muscarinic receptors, since the effects were abolished by pre-treatment with atropine (10( 4) M). Serotonin (5-HT) induced a dose-dependent branchial methysergide-sensitive vasoconstriction which was abolished by pre-treatment with L-NIO, indicating a NO dependent mechanism. On the isolated heart, the NOS inhibitor L-NMMA (N(G) monomethyl-L-arginine) 10(-4) M produced a small, but significant decrease of heart rate and, as a consequence, a decrease of power output, while the NO donor sodium nitroprusside (SNP) 10(-4) M elicited increases of stroke volume, stroke work and power output, suggesting an exogenous NO-dependent positive inotropism. Exposure of the bulbus arteriosus to L-NMMA was without any appreciable effect. In contrast, SNP produced a notable relaxation of the bulbus wall with a marked increase of its stiffness, as indicated by the increase of systolic and diastolic dP/dt max (23 and 104%, respectively). PMID- 12547278 TI - Use of the cytokinesis-block micronucleus assay to measure radiation-induced chromosome damage in lymphoblastoid cell lines. AB - To establish the optimal experimental conditions for the use of the micronuclei (MN) test to determine the level of chromosomal damage induced by ionising radiation (IR) exposure in lymphoblastoid cell lines, a time-course study was performed comparing a normal and an ataxia telangiectasia (AT) cell line, the latter being characterised by an extreme radiation sensitivity. Several parameters were analysed: the use of cytochalasin-B (Cyt-B) to quantify MN, the optimum fixation time to measure radiation-induced MN, the most appropriate treatment dose of IR to distinguish between the normal and the radiosensitive cells and the cell-cycle distribution after irradiation. The results obtained showed that the spontaneous as well as the radiation-induced levels of MN were significantly higher in the AT cell compared to the normal cells (P < 0.001 and P = 0.005, respectively). In both cell types the number of radiation-induced MN were lower in the cultures without Cyt-B than those with Cyt-B (P < 0.001), with the AT cells being distinguished in terms of IR-induced MN from the normal cells only with the addition of Cyt-B. The level of MN formation was independent of the dose of Cyt-B used (3 or 6 microg/ml). The optimum time for radiation-induced MN measured was found to be between 48 and 72 h post-irradiation, with 2 and 4 Gy exposures inducing similar levels of MN. However, as the higher dose caused a greater delay in the cell-cycle, treatment with 2 Gy with MN measurement at 48 h in the presence of 3 microg/ml Cyt-B were chosen as the optimum experimental conditions. This choice was validated using two additional normal and AT cell lines. In conclusion, our results show that the use of Cyt-B increases the sensitivity of the MN test for comparing differences in radiosensitivity between lymphoblastoid cell lines. PMID- 12547279 TI - Induction of micronuclei in mouse and rat by glycidamide, genotoxic metabolite of acrylamide. AB - Male CBA mice and male Sprague-Dawley rats were treated by i.p. injection of glycidamide (GA), the presumed genotoxic metabolite of acrylamide (AA). GA was obtained through a new way of synthesis. As an endpoint of chromosome damage, micronucleus (MN) induction in erythrocytes was measured. Hemoglobin (Hb) adducts were used as a measure of in vivo dose of GA. GA induced linear dose-dependent increases in adduct levels in both species. Rats exhibit, compared with mice, 30% higher Hb adduct levels per unit of administered amount of GA. The incremental MN frequencies per administered dose of GA in mice showed a linear-quadratic dose dependent curve. In the rat no positive dose-response relationship was obtained, probably due to toxic effects to the bone marrow. The main result of this study is the finding that after treatment with synthetic GA the MN frequency per unit of the in vivo dose of GA in the mouse is very similar to that obtained in a previous study, where animals were treated with AA and GA as a metabolite. This equality in potency of GA, whether its in vivo dose is established by injection of synthetic GA or through metabolism of AA, supports the view that GA is the predominant genotoxic factor in AA exposure. PMID- 12547280 TI - Free radical scavengers can differentially modulate the genotoxicity of amsacrine in normal and cancer cells. AB - Amsacrine is an acridine derivative drug applied in haematological malignancies. It targets topoisomerase II enhancing the formation of a cleavable DNA-enzyme complex and leading to DNA fragmentation in dividing cancer cells. Little is known about other modes of the interaction of amsacrine with DNA, by which it could affect also normal cells. Using the alkaline comet assay, we showed that amsacrine at concentrations from the range 0.01 to 10 microM induced DNA damage in normal human lymphocytes, human promyelocytic leukemia HL-60 cells lacking the p53 gene and murine pro-B lymphoid cells BaF3 expressing BCR/ABL oncogene measured as the increase in percentage tail DNA. The effect was dose-dependent. Treated cells were able to recover within a 120-min incubation. Amifostine at 14 mM decreased the level of DNA damage in normal lymphocytes, had no effect on the HL-60 cells and potentiated the DNA-damaging effect of the drug in BCR/ABL transformed cells. Vitamin C at 10 and 50 microM diminished the extent of DNA damage in normal lymphocytes, but had no effect in cancer cells. Pre-treatment of the cells with the nitrone spin trap, N-tert-butyl-alpha-phenylnitrone or ebselen, which mimics glutathione peroxidase, reduced the extent of DNA damage evoked by amsacrine in all types of cells. The cells exposed to amsacrine and treated with endonuclease III and 3-methyladenine-DNA glycosylase II, the enzymes recognizing oxidized and alkylated bases, respectively, displayed greater extent of DNA damage than those not treated with these enzymes. The results obtained suggest that free radicals may be involved in the formation of DNA lesions induced by amsacrine. The drug can also methylate DNA bases. Our results indicate that the induction of secondary malignancies should be taken into account as diverse side effects of amsacrine. Amifostine may potentate DNA-damage effect of amsacrine in cancer cells and decrease this effect in normal cells and Vitamin C can be considered as a protective agent against DNA damage in normal cells. PMID- 12547281 TI - Genotoxicity of lomefloxacin--an antibacterial drug in somatic and germ cells of Swiss albino mice in vivo. AB - The in vivo genotoxicity of lomefloxacin, a diflourinated antibacterial drug, was evaluated by employing mouse in vivo chromosomal aberration test in bone marrow cells and dominant lethal mutation assay in germ cells. Statistically significant reduction in mitotic index, increase in chromosomal aberrations (CAs)/cell and percent abnormal metaphase was observed only at the highest dose (160 mg/kg b.w.) of the drug. In the dominant lethal mutation assay, a statistically significant decrease in the number of implants/female, compared to vehicle control, was noticed only in the females mated with males treated with 32 mg/kg b.w. during the third week of mating, while statistically significant reduction in live implants/female was noticed at both the doses during the second and third weeks of mating. Nevertheless, no significant change in the number of dead implants/female was observed after lomefloxacin treatment. These results seems to indicate that lomefloxacin is a weak clastogen in the bone marrow cells and non mutagenic in the germ cells of mouse in vivo. PMID- 12547282 TI - Phototoxicity and photogenotoxicity of nine pyridone derivatives. AB - Nine structurally related pyridone derivatives were assayed for photogenotoxicity and phototoxicity in the Ames test, the chromosomal aberration test in V79 cells and the neutral red uptake (NRU) test in 3T3 cells. All nine compounds absorb light to a comparable degree at wavelengths between 380 and 430 nm. Seven of the nine compounds were found to produce high quantities of singlet oxygen (1O(2)) upon irradiation in the presence of oxygen. These seven compounds were highly phototoxic in the NRU test, three were clearly and two were marginally photomutagenic in the Ames test, five were assessed as clearly and two as equivocally photoclastogenic in the chromosomal aberration test. Two compounds showed substantially lower 1O(2) yields. The pyridone ring of these two compounds is attached to a non-aromatic ring, while for the seven other compounds the chromophore system including the pyridone ring consists of two or three aromatic rings. One of the two compounds with low 1O(2) yields was distinctly less phototoxic and did not induce photogenotoxic effects. The other, structurally an indolo derivative and not the common thieno derivative, was, however, similarly phototoxic as the seven compounds with high 1O(2) quantum yield and was also clearly photogenotoxic indicating that different action pathways, not involving singlet oxygen, have to be considered at least for this compound. PMID- 12547283 TI - Mutagenicity test system based on a reporter gene assay for short-term detection of mutagens (MutaGen assay). AB - The construction of a bacterial mutation assay system detecting reversions of base substitutions and frameshifts in tetracycline (tet) and ampicillin resistance genes located on low copy plasmids is described. Frameshift mutations were introduced into repetitive GC-sequences and G-repeats known to be mutagenic hot-spots. Base pair substitutions were inserted in or around the active site of the ampicillinase gene thus generating reversibility of the ampicilline sensitivity. The plasmids carry genes to enable sensitive, fast and specific detection of mutagens in bacteria. MucAB was cloned into the test plasmid to enhance error-prone DNA-repair. The conventional reversion principle has been combined with the luminometric measurement of an inducible reporter gene. The revertants are detected after induction of the beta-galactosidase-producing lacZ gene either controlled by its natural lac-promotor or by the more stringently repressed (anhydrotetracyclin inducible) tetA promotor. The tester strains containing the tetA/lacZ reporter gene construct can grow in full medium over the complete assay. This test procedure enables screening for mutations within one working day. Incubation for 16 h reveals high sensitivity. PMID- 12547284 TI - p16 hypermethylation during gastric carcinogenesis of Wistar rats by N-methyl-N' nitro-N-nitrosoguanidine. AB - Inactivation of the tumor suppressor gene, p16 by CpG hypermethylation is a common event in various tumors including gastric carcinoma. The aim of this study is to investigate if p16 hypermethylation is an early and frequent event in gastric carcinogenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). The frequency and timing of p16 hypermethylation during the multistep gastric carcinogenesis in Wistar rats were analyzed in various microdissected gastric lesions. The p16 methylation status and the presence of p16 protein were analyzed by methylation-specific PCR and immunohistochemistry, respectively. Results showed that p16 methylation frequency was correlated with the severity of gastric pathologic lesions, positively. For instance, p16 methylation was found in 2.7% of normal gastric epithelium (n = 36), 16.7% of chronic atrophy gastritis (n = 24), 37.5% of dysplasia (n = 24), 67.4% of gastric adenoma (n = 43), and 85.2% of gastric carcinoma (n = 27). The p16 methylation in the distal stomach epithelium was higher than that in the proximal stomach. p16 protein was expressed in all of 15 p16 unmethylated gastric epithelial samples, but not expressed in all of 12 p16 methylated samples. These results suggest that CpG island hypermethylation may account for the silencing of p16 in rat stomach and is an early event whose accumulation will finally lead to gastric carcinogenesis. PMID- 12547285 TI - A new approach for evaluating in vivo anti-leukemic activity using the SCE assay. An application on three newly synthesised anti-tumour steroidal esters. AB - Three newly synthesised steroidal esteric derivatives of nitrogen mustard (compounds 1-3) were comparatively studied on a molar basis regarding their ability to induce sister chromatid exchanges (SCEs) in normal human lymphocytes in vitro and therapeutic effects on leukemia P388 bearing mice. Compounds 1 and 3 are modified steroidal esters of p-methyl-m-N,N-bis(2-chloroethyl)amino benzoic acid, and compound 2 is a modified steroidal ester of chlorambucil. All compounds induced statistically significant increases in SCEs and decreases in proliferation rate indices (PRIs) of cultured human lymphocytes and significantly increased the life span of P388 bearing mice. In this study, the doses applied for therapeutic purposes upon leukemia P388 bearing mice in vivo were derived from cytogenetic observations in normal human lymphocytes in vitro. A substantially better therapeutic effect was obtained compared to the effect achieved after the use of quite higher doses related with LD(10) values. We have demonstrated that the order of anti-tumour effectiveness of the treatment schedules of the three newly synthesised compounds tested (at doses derived from cytogenetic observations) coincides with the order of the cytogenetic effects they induce. The SCE assay appears to have an application in the clinical prediction of tumour sensitivity to potential chemotherapeutics. PMID- 12547286 TI - The influence of occupational exposure to PAHs on the blood plasma levels of p53 and p21WAF1 proteins. AB - Polycyclic aromatic hydrocarbons (PAHs) are the main source of carcinogenic risk among coke-oven workers. p53 is a tumor suppressor protein that is induced after DNA damage. It regulates the transcription of genes responsible for cell cycle arrest and apoptosis. p21(WAF1) protein is a downstream effector of p53; it induces cell cycle arrest either in the G(1), S, or G(2) phases. It has been shown that carcinogenic PAHs are able to induce the expression of both p53 and p21(WAF1) proteins in vitro. The purpose of the present study was to evaluate the effect of occupational exposure to carcinogenic PAHs on the level of p53 and p21(WAF1) proteins in blood plasma. The exposed group consisted of 66 coke-oven workers (males, average age 41 years, 42% smokers, 58% nonsmokers); the control group consisted of 49 machine workers (males, average age 49 years, 51% smokers, 49% nonsmokers). No difference in the plasma levels of either p53 (using anti-p53 antibody identifying both the mutated and the wild-type form of the protein) or p21(WAF1) protein was found between the exposed and control groups. Smoking had no effect on the levels of either protein in any of the analyzed groups. After stratification of all the subjects into groups according to their exposure to carcinogenic PAHs, a significantly higher level of p53 was found in the group exposed to carcinogenic PAHs <1 microg/m(3) as compared with the group exposed to carcinogenic PAHs >1 microg/m(3). A similar trend was observed for p21(WAF1) protein, even if no correlation between the levels of both proteins was detected. In the overall study a negative correlation between the levels of p53 protein and personal exposure to carcinogenic PAHs was found. These results did not support the expected response. The use of p53 as well as p21(WAF1) protein plasma levels as biomarkers of carcinogenic PAHs exposure requires further studies. PMID- 12547287 TI - Cytogenetic comparison of the sensitivity to mutagens in mice selected for high (HA) and low (LA) swim stress-induced analgesia. AB - Sensitivity to mutagens was studied in mouse lines selectively bred for high analgesia (HA) and for low analgesia (LA) induced by 3-min swimming in 20 degrees C water. Apart from pain-related traits HA mice also manifest, as compared to the LA line, higher emotionality in various behavioural tests, and cope worse with the hypothermic challenge of swimming in cold water. In the present study HA mice appeared more susceptible to the mutagenic effect of whole-body gamma-radiation and mitomycin-C injection. Both treatments caused higher frequencies of chromosomal aberrations and micronucleus in bone marrow cells in the HA than in the LA line. The results are discussed in terms of a genetic correlation between animals' susceptibility to environmental stressors and the mechanism of mutagenesis. As shown by our recent molecular study, the selection for magnitude of swim analgesia has differentiated the parental outbred population into two distinct genotypes characterised by specific minisatellite and microsatellite sequences for each line, which may be genetic markers of particular traits. We conceive that the breeding strategy, along with the differentiation of stress related phenomena, has altered the frequencies of genes controlling DNA repair in each line. PMID- 12547288 TI - Anthocyanins protect against DNA damage induced by tert-butyl-hydroperoxide in rat smooth muscle and hepatoma cells. AB - Anthocyanins are flavonoids present in a variety of pigmented food and, like other flavonoids, seem to play a role in preventing human pathologies related to oxidative stress. In fact, anthocyanins have been shown to exert antiproliferative effects in cell cultures and exhibit antiinflammatory and vasoprotective activities in animal models. Although these biological activities have been related to their antioxidant properties, little is known on the molecular mechanism of action of anthocyanins. The effects of pretreatment with the anthocyanins delphinidin, cyanidin, and their glycoside and rutinoside derivatives against induction of DNA damage induced by tert-butyl-hydroperoxide (TBHP) were evaluated in rat smooth muscle and in rat hepatoma cell lines using alkaline single cell gel electrophoresis (Comet test). In addition, a possible protection exerted by anthocyanins on cell killing, lipid peroxidation, and redox state alterations induced by TBHP was also investigated. It was found that the treatment with TBHP induces the formation of DNA single strand breaks (SSB) and oxidised bases, along with cell killing, lipid peroxidation and redox state alteration. Our data demonstrate that anthocyanins are effective against cytotoxicity, DNA SSB formation and lipid peroxidation induced by TBHP, but they do not have any detectable effect against impairment by TBHP of cellular redox state and on protection against DNA bases oxidation. The presence of a sugar moiety in anthocyanin derivatives reduced this protective effect, mainly in rat hepatoma cells. The different activity of anthocyanins and their derivatives may be explained taking into account a structure/function relationship that could also influence anthocyanin intracellular localisation. PMID- 12547289 TI - Is there a relationship between attention deficit hyperactivity disorder and bipolar disorder? AB - With the increasing recognition of attention deficit hyperactivity disorder (ADHD) in adults and psychotic disorders in children and adolescents, the possibility of a relationship between bipolar disorder (BP) and ADHD has attracted growing interest. This paper critically reviews the scientific literature concerning this postulated relationship by examining evidence from clinico-epidemiological, follow up, family and laboratory studies, including neuroimaging, neuropsychology and genetic studies. The evidence suggests that although the diagnostic categories of BP and ADHD appear to be unrelated, there is support for a possible relationship between some ADHD and manic-like symptoms. However, several fundamental methodological issues require rectification in future research in order to further elucidate the relationship between these disorders. PMID- 12547290 TI - Lithium, anticonvulsants and suicidal behavior in bipolar disorder. AB - BACKGROUND: Lithium has been found to be effective in reducing suicide rates during long term treatment of patients with bipolar disorders. Data on the efficacy of anticonvulsant mood stabilizers in reducing suicide risk are sparse. METHOD: Charts of 140 bipolar patients treated continuously for a minimum of 6 months during a 23-year period of private practice by the senior author were extracted from nearly 4000 patient records. Data extracted from the charts were incidence of completed suicide, number of suicide attempts, and number of hospitalizations for suicidal ideation or behavior per 100 patient-years of either 'on' or 'off' lithium or anticonvulsant mood stabilizer monotherapy. RESULTS: Only one completed suicide (during a period off of lithium) occurred in the patients studied. Incidence of non-lethal suicidal behavior was not different during treatment with lithium, compared with anticonvulsants. Being on a mood stabilizer significantly protected against suicidal behavior. The relative protective effect was more modest than in reports from other treatment settings. LIMITATIONS: This was a retrospective chart review study of naturalistically treated patients. CONCLUSIONS: Treatment of patients with bipolar disorder with either lithium or anticonvulsant mood stabilizers was associated with reduced risk of suicidal behavior. This study did not find evidence for a difference in the protective effect of the two types of mood stabilizing medications against non-lethal suicidal behavior in the naturalistic setting of private practice. PMID- 12547291 TI - A follow-up study of DSM-III-R generalized anxiety disorder with syndromal and subsyndromal major depression. AB - OBJECTIVE: The authors examined the long-term outcome of generalized anxiety disorder with depressive symptoms utilizing both categorical and dimensional analyses. METHOD: Thirty-nine out-patients with a DSM-III-R diagnosis of generalized anxiety disorder (GAD) with depressive symptoms, both with (n=23) and without (n=16) syndromal major depression (MD) participated in an 11-week clinical trial. Approximately 18 months after initial screening, these individuals were once again evaluated using a structured diagnostic interview and a battery of rating scales. RESULTS: Three distinct groups were discernible at follow-up. Twenty-three (60%) of the patients remained syndromal for GAD; 10 patients (43%) were in partial remission from GAD; six (15%) were asymptomatic. Of the 23 patients who were syndromal for MD at baseline, 13 (56%) remained syndromal for MD at follow-up. All of the patients who were syndromal for MD at follow-up remained syndromal for GAD as well. CONCLUSIONS: Outcomes in this study were quite divergent, though they support the concept of GAD as a chronic illness in most patients, with or without MD. The presence or absence of MD versus subsyndromal depression at baseline appeared to have relatively little impact upon the outcome. Patients with subsyndromal anxiety and depressive symptoms may be at special risk for syndromal disorders over time. PMID- 12547292 TI - Relapse of major depression after complete and partial remission during a 2-year follow-up. AB - BACKGROUND: Rates of remission and relapse were studied over more than 2 years in a sample of Spanish outpatients with DSM-III-R criteria of unipolar major depressive episodes. METHODS: Patients were treated following standardised pharmacological protocols at our centre. In the first visit, the structured clinical interview for DSM-III-R (SCID) was used. The following visits were held monthly. Phases of evolution were recorded using the Hamilton Depression Rating Scale (HDRS), applying the Frank criteria. RESULTS: A significantly greater proportion of relapse was observed in the partial remission group compared to the complete remission one. The rate of relapses for patients in complete remission was 15.18%, while for patients in partial remission was 67.61%. Partial remission was significantly associated with relapses. LIMITATIONS: The short duration of the study and the decreasing sample size during the follow-up. CONCLUSIONS: Partial remission after a depressive episode seems to be strongly associated with relapses. Moreover, this clinical factor could by itself fully predict short-term relapses. CLINICAL RELEVANCE: The study shows the importance of reaching complete remission to decrease the rate of short-term relapses. PMID- 12547293 TI - Severity of stressful life events in first and subsequent episodes of depression: the relevance of depressive subtype. AB - BACKGROUND: Recent studies have reignited debate concerning the relationship between stressful life events and depressive subtypes, particularly in relation to first versus subsequent episodes. AIMS: To investigate the relationship between stressful life events and variably defined melancholic/non-melancholic depressive subtypes, and the import of such life events to first compared with subsequent episodes across those subtypes. METHOD: Acute and chronic stressful life events were rated in 270 patients with DSM-IV Major Depressive episodes who were allocated to melancholic and non-melancholic groups separately as defined by DSM-III-R, DSM-IV, the Newcastle criteria and the CORE system. RESULTS: Severe stressful life events (both acute and chronic)-as defined by DSM-III-R axis IV were more likely to occur prior to first rather than subsequent episodes, particularly for those with non-melancholic depression. LIMITATIONS: Dependence or independence of life events was not assessed. Genetic vulnerability to depression was not determined. Life events in first and subsequent depressive episodes were compared cross-sectionally between groups, not prospectively in the same cohort of patients. There were no differences in the number of severe life events-as defined by clinician consensus-between the first and subsequent episodes. CONCLUSIONS: These findings are consistent with other studies in suggesting an enhanced sensitisation of depressed patients to subsequent episodes of depression, but suggest that any such phenomenon is specific to non melancholic depression, in comparison to one key previous study. PMID- 12547294 TI - Theory of mind deficits in bipolar affective disorder. AB - BACKGROUND: Bipolar affective disorder patients often show cognitive deficits that are similar to those found in schizophrenia patients. Theory of mind (the ability to understand others' mental states) is compromised in currently ill schizophrenia patients. This study aimed to establish whether similar deficits are found in bipolar patients. METHODS: We measured theory of mind ability in 20 bipolar-manic patients, 15 bipolar-depressed patients, 13 bipolar patients in remission and 15 normal controls. The task, which controlled for memory and comprehension, had previously been used in a study of schizophrenia patients. RESULTS: Impaired performance on theory of mind was found for both bipolar depressed and bipolar-manic patients, even when memory was controlled for. No impairment was observed in the remitted patients. LIMITATIONS: The manic patients scored lower than the remitted patients on a brief measure of intelligence; no other group differences in IQ were significant. CONCLUSIONS: Theory of mind deficits are found in currently symptomatic bipolar patients. These findings add to growing evidence that common mechanisms may contribute to bipolar affective disorder and schizophrenia. PMID- 12547295 TI - Increased risk of developing dementia in patients with major affective disorders compared to patients with other medical illnesses. AB - BACKGROUND: The association between affective disorder and subsequent dementia is unclear. Our aim was to investigate whether patients with unipolar or bipolar affective disorder have an increased risk of developing dementia compared to patients with other chronic illnesses. METHOD: By linkage of the psychiatric and somatic nation-wide registers of all hospitalised patients in Denmark, 2007 patients with mania, 11741 patients with depression, 81380 patients with osteoarthritis and 69149 patients with diabetes were identified according to diagnosis at first-ever discharge from a psychiatric or somatic hospital between 1 January 1977 and 31 December 1993. The risk of receiving a diagnosis of dementia on subsequent re-admission was estimated with the use of survival analyses. RESULTS: Patients with unipolar or bipolar affective disorder had a greater risk of receiving a diagnosis of dementia than patients with osteoarthritis or diabetes. Differences in age and gender and the effect of alcohol- or drug-abuse did not explain these associations. CONCLUSION: Patients with unipolar or bipolar affective disorder seem to have an increased risk of developing dementia compared to patients with other illnesses. LIMITATION: The study includes only patients who have been hospitalised at least once. CLINICAL RELEVANCE: Patients with unipolar or bipolar affective disorder may be at increased risk of developing dementia. PMID- 12547296 TI - Evidence for a late onset bipolar-I disorder sub-group from 50 years. AB - BACKGROUND: Age of onset has been used to identify aetiological sub-groups in complex inherited disorders such as Alzheimer's disease and osteoarthritis. We examined the relationship between age of onset and family history in bipolar-I disorder in an attempt to identify subgroups. METHODS: All patients discharged from a district in-patient service diagnosed with bipolar disorder in a 7-year period were ascertained from a case register (n=277). Diagnosis by DSM-IIIR criteria was confirmed; family history and age of first admission were recorded from case notes. RESULTS: Age of first admission in those with a negative family psychiatric history was significantly older (P=0.029) with a skewed age distribution. This non-familial group contained significantly more subjects with first admission from the age of 50 years (P=0.007). CONCLUSIONS: Patients with bipolar-I disorder whose age of first admission is 50 years or above may belong to a different aetiological sub-group. PMID- 12547297 TI - Possible effect of gender and season on the length of hospitalisation in unipolar major depressives. AB - BACKGROUND: Seasonal variation as well as gender differences of several phenomena of affective disorders are a common topic of interest. METHODS: The authors analysed the possible effect of season and gender on the length of hospital stay in 529 in-patients with unipolar major depressive episode. RESULTS: Age and menopausal status alone did not influence the length of hospitalisation but there was a statistical tendency (only for females) for the shortest hospital stay in summer, that reached significance in females younger than 50 years. CONCLUSIONS: The results suggest a possible seasonal and gender effect on recovery from major depression. LIMITATIONS: Retrospective nature of the study, lack of systematic assessment of clinical response and no data collection about marital status and living conditions, that also can influence the time of discharge. PMID- 12547298 TI - P300 changes in major depressive disorders with and without psychotic features. AB - BACKGROUND: Although there are many P300 studies in depressive patients, only a few studies have focused on the effects of psychotic features in depression and of response to antidepressant treatment on P300. This study was designed to investigate possible differences in the P300 component of event-related potentials in depressed patients with and without psychotic features and if any, to see whether these changes altered with treatment of depression. METHODS: Thirty-six patients with major depressive disorder diagnosed according to DSM-IV, and 20 healthy control subjects were involved in the study. Sixteen of the patients had psychotic features. Auditory P300 was recorded before treatment and after remission. RESULTS: Pretreatment P300 latencies were significantly prolonged both in patients with and without psychotic features compared to controls. Pretreatment P300 amplitudes were significantly decreased only in the patients with psychotic features. After treatment of depression, delayed P300 latencies in both patient groups and decreased P300 amplitude in the patient group with psychotic features were normalized. LIMITATIONS: The medication status of the patient was heterogeneous. CONCLUSION: Since the impairment seems to be improved by drug treatment, prolonged P300 latency might be a state marker for a major depressive episode, and decreased P300 amplitude which is correlated with paranoid ideation might be more associated with psychotic subtype. PMID- 12547299 TI - The development of a six-item daily self-report measure assessing identified depressive domains. AB - BACKGROUND: There is debate as to whether any anti-depressant strategy acts more rapidly than any other, while 'improvement' in overall depression severity reflects a summation of individual domains that may individually show differing trajectories. We developed a brief self-report measure of depression's constituent constructs to allow such issues to be examined. METHODS: A 25-item measure was prepared and completed daily by depressed patients until they had evidenced distinct improvement. RESULTS: Factor analyses favoured a six-factor structure, with constructs labelled depression, irritability, brooding, poor concentration, insomnia and anxiety. Scores on those constructs were differentially associated with overall depression severity as measured on the clinician-rated Hamilton measure, arguing for their potential utility. LIMITATIONS: The original data set was small while the measure's utility will only emerge in application studies. CONCLUSIONS: The six-item measure is appended and several possible application studies noted. In particular, we favour its evaluation in studies examining the impact and time to onset of differing antidepressant strategies on differing depressive sub-types. PMID- 12547300 TI - Bipolar II depressive mixed state changes with age. PMID- 12547302 TI - Antiepileptic drugs in schizophrenia: a review. AB - The first choice group of psychotropic agents in schizophrenia is neuroleptics. However, this treatment is not effective in all patients and with every symptom. We summarize papers published on the role of antiepileptic drugs in treatment resistant schizophrenia. We have searched the computer database system MEDLINE for relevant articles including reviews, reports of drug studies and case histories. Antiepileptic drugs can change symptoms of schizophrenia by their action on GABA-ergic neurotransmission or via anti-glutamatergic mechanisms. High doses of adjunctive benzodiazepines reduce positive symptoms, anxiety, and agitation. Carbamazepine is effective in affective symptoms of schizophrenia and influences violent behavior in psychotic patients. Its anti-kindling action may represent a promising treatment strategy for some patients with chronic course of schizophrenia. Valproate treatment leads to a decrease in positive symptoms as well as hostility. Lamotrigine is expected to influence the positive, negative, affective, and cognitive symptoms of schizophrenia. New antiepileptics (e.g., gabapentin, oxcarbazepine, topiramate, vigabatrin) present a promise as potential adjuncts to neuroleptic treatment in resistant symptoms of schizophrenia. PMID- 12547303 TI - Temperament in schizophrenia: a study of the tridimensional personality questionnaire (TPQ). AB - PURPOSE: The aim of this study was to assess dimensions of temperament as defined by Cloninger's neurobiological model using the tridimensional personality questionnaire (TPQ) in a sample of consecutively recruited schizophrenic patients. SUBJECTS AND METHODS: We used the French version of the TPQ to compare 45 stable, euthymic schizophrenic patients with 126 controls with no personal or familial history of psychiatric disorder. After comparison of TPQ scores between groups, we also performed a multivariate analysis to avoid the confounding effects of age, gender, and alcohol and substance use disorder comorbidity. RESULTS: Harm avoidance (HA) was higher in schizophrenic patients than in controls. DISCUSSION AND CONCLUSION: This replicates and extends the results of previous studies suggesting that schizophrenic patients have high HA and that HA might be a marker for underlying genetic vulnerability to schizophrenia. PMID- 12547304 TI - Self-experienced vulnerability, prodromal symptoms and coping strategies preceding schizophrenic and depressive relapses. AB - For the first time, the present study explores pre-episodic disturbances, i.e. self-experienced vulnerability and prodromal symptoms, and related coping strategies preceding schizophrenic and depressive relapses. After complete recovery from the acute episode, 27 patients with recurrent schizophrenic and 24 patients with recurrent depressive episodes were assessed retrospectively for pre episodic disturbances and related coping strategies with the "Bonn scale for the assessment of basic symptoms-BSABS". All (100%) of the schizophrenic and 23 (96%) of the depressive patients showed pre-episodic disturbances. Patients with schizophrenia showed significantly more often an increased emotional reactivity and certain perception and thought disturbances. Depressive patients reported significantly more often an impaired tolerance to certain stress and disorders of emotion and affect. Sixty-three percent of the schizophrenics and 87% of the depressives reacted to pre-episodic disturbances with coping strategies. The pre episodic disturbances in patients with schizophrenia could be described in terms of mild psychotic productivity, those in depressives in terms of mild depressive syndrome. Future studies will have to show if these findings can be replicated in first episode or initial prodromal state samples and if the assessment of mild psychotic productivity and mild depressive syndrome can be used for early diagnosis and early intervention in schizophrenia and depression. PMID- 12547305 TI - Clinical and psychosocial outcome of patients affected by panic disorder with or without agoraphobia: results from a naturalistic follow-up study. AB - OBJECTIVE: A clinical and psychosocial follow-up study of a cohort of 85 patients affected by panic disorder (PD) with or without agoraphobia was performed an average of 40 months after initial observation and following a mean duration of illness of 8 years. METHODS: Eighty-five out of 130 patients affected by PDs with or without agoraphobia according to DSM-III R criteria, examined between 1990 and 1995 at an outpatient clinic were re-examined in 1997/1998 using the same standardized clinical evaluation performed on admission. Patients also underwent a structured diagnostic interview (Mini International Neuropsychiatric Interview, MINI) and psychosocial evaluation (Scale of Sheehan's Disability Scale, DISS, Baker and Intagliata's Satisfaction with Life Domains Scale, SLDS). RESULTS: At follow-up, the percentage of patients who had either improved or were in remission was considerably higher among those initially diagnosed as PD with respect to those diagnosed as panic disorder with agoraphobia (PDA): Thirty-eight percent of PD and 20.6% of PDA patients were in clinical remission. Mild panic symptoms and phobic avoidance were found in the majority of patients who were still symptomatic (respectively 71% and 57%). Approximately 60% of patients reported a significant difficulty in performing daily activities and 40% expressed dissatisfaction in at least 50% of life domains considered. Seventy-two percent of subjects examined were still undergoing pharmacological treatment at the time of follow-up. CONCLUSIONS: The findings of the study are suggestive of a chronic illness with a significant impact on everyday quality of life of patients. PMID- 12547306 TI - Residual symptoms and comorbidity in panic disorder. AB - BACKGROUND: The aim of this study was to assess the outcome of the comorbid conditions of panic disorder after 1 year of treatment, emphasizing the detection of residual symptoms and their relationship to other clinical variables. METHODS: Subjects (N = 64) were assessed by the Structured Clinical Interview for DSM-III R and the Eysenck Personality Questionnaire. Comorbidity with other disorders, scores on Hamilton Anxiety Rating Scale and Hamilton Depression Rating Scale were assessed at baseline and after 12 months. Criteria for residual anxiety/somatic symptoms were defined. RESULTS: Reduction in generalized anxiety disorder rates accounted for a significant decrease in comorbidity at 1-year follow-up, with regard to baseline assessment. When the more severe symptoms of the disorder had remitted, a third of the patients referred physical symptoms with some concern over a fluctuating state of anxiety. The said symptoms were neither a recurrence of panic disorder nor did they account for other anxiety or somatoform disorders. Lower scores on extraversion predict higher risk of residual symptoms. DISCUSSION: The persistence of residual anxiety/somatic symptoms in a third of the patients who apparently achieved a good response to treatment of panic disorder might characterize a minor form of chronic persistence of this condition. CONCLUSIONS: The subgroup of patients with residual symptoms would not be detectable by follow-up studies, which focus on the assessment of relapse of panic disorder by means of strictly defined diagnostic criteria. PMID- 12547307 TI - Insight in obsessive-compulsive disorder: a study of an Italian sample. AB - Insight is a complex phenomenon that can be interpreted according to a dimensional model. Given the controversial data of insight in obsessive compulsive disorder (OCD), our study aimed to investigate insight in an Italian sample of patients with OCD by means of the specific item on the Yale-Brown obsessive-compulsive scale (Y-BOCS) and to explore the possible correlations between it and clinical features. One hundred and seventeen out-patients with a DSM-IV diagnosis of OCD and different comorbid psychiatric disorders were included in the study and assessed by means of the Y-BOCS, Hamilton rating scale for depression (HRSD) and the global clinical impression. The results showed that almost 50% of the patients had an excellent level of insight and 15% had a little or no insight. No correlation between levels of insight and clinical features was observed, except for a negative trend with the presence of somatic obsessions. In addition, a trend towards a lower level of insight was observed in those bipolar patients with a positive history of repeated manic or hypomanic episodes. Further studies seem to be necessary in order to establish whether or not OCD patients with poor insight represent a distinct sub-group of patients. PMID- 12547308 TI - No relationship between testosterone levels and depressive symptoms in aging men. AB - We investigated the associations between depression and serum testosterone levels in the elderly. There was no significant difference in the mean levels of the serum testosterone between the groups of patients and normals. For both groups, there was no significant correlation between testosterone and total depression scores or age. PMID- 12547309 TI - Psychotropic prescription in non-psychiatric hospital settings. AB - A study was conducted to assess differences in psychotropic prescription (PP) in various non-psychiatric hospital settings. After adjustment for demographic, medical and psychological status, rates of PP were significantly lower for surgical, intensive care and outpatients and higher for geriatric patients than for patients in other settings, suggesting inadequate consideration of psychiatric problems in certain contexts, in particular intensive care units. PMID- 12547310 TI - Cardiac risk at the onset of treatment in patients treated with benzodiazepines and clozapine. PMID- 12547311 TI - Olanzapine and improvement of tardive dyskinesia. PMID- 12547326 TI - Emissions of polychlorinated dibenzo-p-dioxins and dibenzofurans from stack flue gases of sinter plants. AB - This study investigated four sinter plants, classified into two categories--with selective catalytic reduction (SCR) (S1, S2 and S3) and without SCR (S4) as its air pollution control device. Polychlorinated dibenzofurans (PCDFs) are dominant in the stack flue gases of sinter plants with and without SCR. The polychlorinated dibenzofurans/polychlorinated dibenzo-p-dioxins (PCDFs/PCDDs) ratio exceeds 1 extremely. The degree of chlorination of the sinter plant without SCR is towards hepta and hexa congeners while that of the sinter plant with SCR is towards penta and hexa congeners. PCDD/Fs are indeed decomposed (75.5% and 69% on ng and ng TEQ bases, respectively) and not only reduced in degree of chlorination. The overall concentration in the stack flue gas of sinter plants with SCR (7.97-14.1 ng/Nm(3); 0.995-2.06 ng TEQ/Nm(3)) is lower than that of the sinter plant without SCR (28.9 ng/Nm(3); 3.10 ng TEQ/Nm(3)). In Taiwan, based on the emission factors of 0.970 microg TEQ/ton-feedstock (sinter plants with SCR) and 3.13 microg TEQ/ton-feedstock (sinter plant without SCR), the annual PCDD/F emission of 44.7 g TEQ/year from sinter plants is 60 and 121 times higher than those from municipal solid waste incinerators (0.750 g TEQ/year) and MWIs (0.369 g TEQ/year). These results show that sinter plants have become the dominating PCDD/F emission source since tighter emission limits have been applied to incinerators. PMID- 12547327 TI - PCBs, PCNs, PCDD/Fs, PAHs and Cl-PAHs in air and water particulate samples- patterns and variations. AB - Methodology for the determination of biologically active polychlorinated biphenyls (PCBs), non-ortho PCBs, polychlorinated naphthalenes (PCNs), polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), polycyclic aromatic hydrocarbons (PAHs) was used to investigate concentrations and patterns of certain chlorinated PAH (Cl-PAH) in source related samples using synthetic reference mixtures. Thus, in addition to the above mentioned compounds, mono heptachlorosubstituted fluorenes, phenanthrenes/anthracenes and pyrenes/fluoranthenes (Cl-PAHs) were measured in vapour and particulate air samples from urban road tunnels, samples of settling particulate matter (SPM), and in bottom sediment samples from two point source locations (pulp and paper, and Mg-plant/Fe-Mn-smelter/chlor-alkali) and in the Baltic Sea. Concentrations in air samples followed: PAHs>PCBs>PCNs>non-ortho PCBs or Cl-PAHs>PCDD/Fs. SPM samples collected at increasing distance to the urban area of Stockholm showed: PAHs>PCBs>PCNs>PCDD/Fs>non-ortho PCBs or Cl-PAHs. For all compound groups there was a tenfold (Cl-PAHs fivefold) concentration decrease in SPM samples from highest levels in the urban water area to lowest levels at a distance of 26 km from city centre. PCB profiles of SPM showed similarities with combined profiles of Aroclor 1242 and 1254. PCN profiles of SPM showed similarities with combined profiles of Halowax 1099 and 1014. A correlation with concentration of all tested Cl-PAH and their corresponding parent PAH was found only for Cl-fluorene. PMID- 12547328 TI - Isomer specific synthesis using the Suzuki-coupling. Polychlorinated terphenyls as standards for environmental analysis. AB - Defined polychlorinated terphenyl (PCT) single congeners as reference standards are the prerequisite for the development of analytical methods for their determination and quantification in the environment. The selective synthesis of PCTs for environmental analytical purposes by application of the Suzuki-coupling reaction is described. Under easily modified standard reaction conditions of this coupling process the PCTs can be obtained by reaction of benzeneboronic acids with dibromobenzenes mostly in good yields, as described by the synthesis of following PCT congeners: p-PCT (3,3("),5,5(")-tetrachloro-, 2,2("),4,4(") tetrachloro-, 2('),3,3("),5,5(")-pentachloro-); m-PCT (3,3("),5,5(")-tetrachloro ) and o-PCT (3,3("),5,5(")-tetrachloro-). The terphenyl congeners were characterized by NMR (1H, 13C)- and FT-IR-spectroscopy. Their purity was checked by GC/MS analysis. The experimental and quantum-chemically calculated FT-IR spectra were compared and it was shown, that the determination of the chlorine substitution pattern in the terphenyl congeners by their typical absorption spectra is possible. PMID- 12547329 TI - Optimization of the purification method for dioxin analysis in human serum and temporal changes in background dioxin levels in the general population. AB - To perform high quality and high throughput measurements, complicated purification procedure by two rounds of open column chromatography was simplified by appraising the components and adding a sufficient amount of adsorbents to the column. The procedure consists of two open column techniques using a multi-layer silica gel column and an active carbon dispersed silica gel column. This purification procedure is used in "Standard manual for dioxin analysis in human blood" published in 2000 (Ministry of Health and Welfare, Japan, 2000). Consequently, a purification procedure using only 1 g of 10% AgNO(3)/silica gel can remove the matrix of blood serum as effectively as a multi-layer silica gel column. In addition, 0.1 g of carbon silica gel, equivalent to 1/10 the weight used currently in analyses, separates and recovers the dioxins in purified extract of blood serum. Furthermore, changes in the background level in the general population were clarified by monitoring the dioxin concentrations in human blood. The dioxin concentrations and ratios in blood in the general population remained at a tolerable level for three months. Therefore, the blood serum was available for the detection of a remarkable elevation in dioxins level considered to be indicative of contamination due to high-density exposure. PMID- 12547330 TI - Estrogenic effects of phenolic compounds on glucose-6-phosphate dehydrogenase in MCF-7 cells and uterine glutathione peroxidase in rats. AB - In this study, we tested phenolic compounds such as bisphenol A (BPA), 4 nonylphenol (NP), 4-octylphenol (OP) and 4-propylphenol (PP) by using glucose-6 phosphate dehydrogenase (G6PD) in estrogen sensitive human breast cancer cells (MCF-7 cells) and glutathione peroxidase (GPx) in female immature Sprague-Dawley (SD) rats. This study was designed to investigate whether phenolic compounds have estrogenic effects in these useful screening methods for endocrine disruptors. We chose 6 h as the incubation period for the G6PD assay through a preliminary experiment using 17beta-estradiol (E2). Above the concentration of 1 x 10(-8) M, BPA significantly increased the G6PD activity in a concentration-dependent manner, relative to the control. NP (over the concentration of 1 x 10(-9) M) also enhanced the G6PD activity by about 1.8 times that of the control. OP produced weaker effects on G6PD than NP, and showed a tendency to increase the G6PD activity. PP did not affect the G6PD activity. These results show that BPA and NP have the effect of enhancing G6PD activities in MCF-7 cells. In the in vivo GPx assay, both BPA and E2 significantly increased the uterus wet weights and dramatically enhanced uterine GPx activities in immature female rats in a dose dependent manner. Treatment with NP (500 mg/kg/day) increased significantly both the uterine GPx activity and the uterus wet weights in immature female rats. OP (500 mg/kg/day) also caused a significant increase in uterine GPx activity, but had no effect on the uterus wet weights. This finding indicates that the change in uterine GPx activities could be a more sensitive parameter than that of uterus wet weights in immature rats. This study implies that phenolic compounds have a weak estrogenic effects. PMID- 12547331 TI - Improvements in dioxin abatement strategies at a municipal waste management plant in Barcelona. AB - This study presents the results of a dioxin abatement programme undertaken in the municipal waste incineration plant of Montcada i Reixac (Barcelona, Spain) after the replacement of an obsolete air cleaning device by a new flue gas treatment system. A number of sampling campaigns were conducted with the aim of characterising stack gas emission levels of polychlorinated dibenzo-p-dioxins (PCDDs)/polychlorinated dibenzofurans (PCDFs) and to evaluate initial specifications of dioxin stack gas emission values below 0.1 ng I-TEQ/Nm(3). Preliminary results revealed levels between 44 and 111 ng I-TEQ/Nm(3) when the gas-cleaning system consisted only of an old electrostatic precipitator (ESP). Decreased levels around 15 ng I-TEQ/Nm(3) were observed when the semi-dry scrubber began to operate and the ESP was switched off. Again, remarkable dioxin removal was observed after the installation of the fabric filter and levels around 0.3-0.4 ng I-TEQ/Nm(3) were soon achieved. Nevertheless, the limit of 0.1 ng I-TEQ/Nm(3) was reached by additional injection of activated carbon which helped to lower PCDD/PCDF levels to around 0.036 ng I-TEQ/Nm(3). The results also demonstrated a significant change in the dioxin distribution present in combustion-derived materials (stack gas emission, bottom ash and solid waste from gas treatment). The major dioxin fraction was found in gaseous matrices before the flue gas control system was upgraded. After this step, the major dioxin fraction content was observed in solid waste from gas treatment. PMID- 12547332 TI - Quantification of PCDD/F concentrations in animal manure and comparison of the effects of the application of cattle manure and sewage sludge to agricultural land on human exposure to PCDD/Fs. AB - PCDD/Fs were quantified in samples of cattle, pig, sheep and chicken manure. TEQs ranged from 0.19 ng TEQ/kg dw for the pig manure to 20 ng TEQ/kg dw for one cattle manure sample. Exposure assessments were carried out to compare the effect of various sewage sludge and cattle manure land application scenarios on human PCDD/F exposure. Background exposure was estimated at 4.9 pg TEQ/kg bw/day and it was found that land application of sewage sludge and cattle manure at contemporary rates did not increase PCDD/F TEQ background exposure. It was concluded that the application of sewage sludge, under the new UK guidelines, and cattle manure to agricultural land, do not make a significant contribution to human exposure to PCDD/Fs. PMID- 12547333 TI - Human exposure to dioxins through the diet in Catalonia, Spain: carcinogenic and non-carcinogenic risk. AB - The main objectives of this study were to estimate the dietary intake of dioxins by the population of Catalonia, Spain, to determine which food groups showed the greatest contribution to this intake, and to assess the health risks potentially associated with the dietary dioxin intake. From June to August 2000, food samples were randomly acquired in seven cities of Catalonia. Dioxin concentrations were determined in 108 samples belonging to the following groups: vegetables, fruits, pulses, cereals, fish and shellfish, meats and meat products, eggs, milk and dairy products, and oils and fats. Estimates of average daily food consumption were obtained from recent studies. Total dietary intake of dioxins for the general population of Catalonia was estimated to be 95.4 pg WHO-TEQ/day (78.4 pg I-TEQ/day), with fish and shellfish (31%), diary products (25%), cereals (14%) and meat (13%) showing the greatest percentages of contribution to dioxin intake. The contribution of all the rest of food groups to the total dietary intake was under 20%. The non-carcinogenic risk index of dioxin intake through the diet was in the range 0.34-1.36, while the carcinogenic risk level was 1,360 excess cancer over a lifetime of 70 years. Our results corroborate the decreasing tendency in dietary intake of dioxins found in recent studies (2000-2001) from various countries. PMID- 12547334 TI - PCDD/Fs and PCBs in Baltic herring during the 1990s. AB - Baltic herring samples caught from the Baltic Sea during the spring periods of 1993-1994 and 1999 were analysed for polychlorinated dibenzo-p-dioxins (PCDD), polychlorinated dibenzofurans (PCDF), and polychlorinated biphenyls (PCB). For analyses, 1570 individual herring were combined to 120 pools. Correlations between concentrations of congeners 1,2,3,7,8-PeCDD, 1,2,3,7,8-PeCDF, 2,3,4,7,8 PeCDF, and 2,3,4,6,7,8-HxCDF, and age of herring were the strongest (r>0.8) followed by correlations between PCB congeners PCB 105, 118, 126, 156, 169 and 180 (r>0.7), and age of herring. Due to higher fat percentage in herring in the Gulf of Bothnia the concentrations of PCDD/Fs and PCBs on fresh weight (fw) basis were higher than in herring in the Gulf of Finland. The concentrations of WHO(PCDD/F)-TEQs ranged from 1 to 27 pg/g fw, depending on the age and catchment area of herring, and concentrations of WHO(PCB)-TEQs reached 32 pg/g fw. Between the two studied time points no clear downward trend in concentrations was observed. PMID- 12547335 TI - Photodegradation of PCDD/Fs adsorbed on spruce (Picea abies (L.) Karst.) needles under sunlight irradiation. AB - Spruce (Picea abies (L.) Karst.) needles were exposed to exhaust gas containing polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) generated by combustion of polyvinyl chloride, wood, high-density polyethylene, and styrene. Photodegradation of PCDD/Fs adsorbed on spruce needles under sunlight irradiation was studied. The photodegradation of PCDD/Fs follows pseudo-first-order reaction kinetics, with photolysis half-lives ranging between 40 and 100 h. The photolysis rates of PCDF congeners are higher than PCDD congeners with the same chlorinated substitutions. Higher chlorinated PCDD/Fs tend to photolyze slowly. The wax components in spruce needles may act as proton donors and accelerate the photolysis rate. C-Cl cleavage through the addition of protons to PCDD/F molecules may be an important route for PCDD/Fs photodegradation. PMID- 12547336 TI - Fate of 1,2,3,4,6,7,8-heptachlorodibenzofuran and pentachlorophenol during laboratory-scale anaerobic mesophilic sewage sludge digestion. AB - The possibility of the formation of PCDDs and dechlorination of PCDFs during the anaerobic digestion of sewage sludge in laboratory scale digesters was investigated. Digesters were spiked with 1,2,3,4,6,7,8-HpCDF-13C(6) (240 ng/g organic matter (OM)) and pentachlorophenol (PeCP)-13C(6) (24 microg/g OM) and the output sludge monitored for 60 days. No dechlorination or formation of the labelled or native PCDD/Fs was observed. The detectable 1,2,3,4,6,7,8-HpCDF 13C(6) dechlorination yield was 0.0008-1% depending on homologue group and the detectable formation of OCDD yield was 0.00042% PeCP-13C(6). Preferential respiration of other, more bioavailable, substrates is suggested as the most plausible explanation for the lack of dechlorination. Formation of PCDDs from PeCP has been observed in aerobic environments but this study provides further evidence that it is not a precursor for formation during anaerobic digestion. PMID- 12547337 TI - Formation behavior of PCDD/Fs in PVC pyrolysis with copper oxide. AB - Formation and decomposition behaviors of PCDD/Fs during pyrolysis of polyvinyl chloride (PVC) with CuO have been investigated. These reactions proceed simultaneously, and the rate of decomposition exceeds that of formation with further retention. More 2,3,7,8-TCDD is formed when the dechlorination of PCDD/Fs proceeds significantly. Homologue profile patterns of PCDD/Fs show that the fractions of O8CDD and H6CDFs are relatively larger within PCDDs and PCDFs, respectively. Extremely large amounts of PCDD/Fs are obtained with the long retention time at 200 degrees C. The formation of PCDD/Fs decreases drastically with increase in the molar ratio of CuO/PVC. The acceptability of thermodynamic calculations on the formation of PCDD/Fs is also investigated. The thermodynamic calculated tendency of the effect of oxygen on the formation of PCDD/Fs agrees well with the experimental results, although absolute values of the amount of PCDD/Fs are much different. PMID- 12547338 TI - Fate of isoxaben in a containerized plant rhizosphere system. AB - Commercial production of ornamental plants is an important industry in the United States and involves a complex technology that includes the use of herbicides. Isoxaben[N-[3-(1-ethyl-1-methylpropyl)-5-isoxazolyl]-2,6-dimethoxybenzamide] is a pre-emergence herbicide used for controlling weeds in many areas including containerized ornamental plants. Degradation was studied in potting mix (80% bark, 20% sand) with three different regimes (sterile, bulk and rhizosphere). The rhizosphere regime contained Switch Grass (Panicum virgatum), and plants were allowed to grow for 14 days before adding isoxaben (10 microg/g potting mix). Isoxaben was degraded to 0.5 microg/g in 60 days giving a half-life of 7 days. Two degradation products were detected: 3-nitrophthalic acid in the rhizosphere and bulk regimes and 4-methoxyphenol in the sterile regime. Microbial population shifts were determined by fatty acid methyl ester profile analysis and were influenced by the introduction of a plant (rhizosphere regime) and by isoxaben addition. PMID- 12547339 TI - Irreversible binding of o,p'-DDD in interrenal cells of Atlantic cod (Gadus morhua). AB - Precision-cut tissue slices of the anterior kidney from Atlantic cod (Gadus morhua) were prepared with a Krumdieck tissue slicer and exposed to 2-(2 chlorophenyl)-2-(4-chloro-(14C)phenyl)-1,1-dichlorethane (o,p(')-[14C]DDD) in vitro. Microautoradiography revealed irreversible o,p(')-DDD-derived binding confined to the glucocorticoid producing interrenal cells (adrenocortical analogues). This cell-selective binding was confirmed by means of autoradiography at different levels of resolution on Atlantic cod administered o,p(')-[14C]DDD intragastrically. The results provide evidence for a site-specific metabolic activation and irreversible binding of o,p(')-DDD in the interrenal cells, which, in turn, may modify glucocorticoid homeostasis. PMID- 12547340 TI - Hydrodehalogenation of chlorobenzene on activated carbon and activated carbon supported catalysts. AB - Chlorinated aromatic compounds in (waste) gases can be removed and/or dehalogenated by passing over a bed of activated carbon (AC) in a hydrogen containing atmosphere. Dehalogenation of the model compound chlorobenzene (PhCl) to HCl is complete at 490 degrees C--rather than the approximately 900 degrees C needed for the mere gas-phase reaction--but part of the benzene moieties is retained on the AC, resulting in its rather rapid deactivation, apparently due to a large decrease in surface area. Therefore, <1 mmol PhCl per gram of 'catalyst' could be processed. At 600 degrees C chlorobenzene yields 100% of both HCl and benzene, but still the AC lost most of its activity in time. Performances are compared of three different commercial ACs. As a HCl-washed AC is just as active, metals are not involved. This acid-washed AC was used as a support for 10 wt% Ni or Fe catalysts. While Fe did not show appreciable activity, results with Ni are promising. The yields of HCl and benzene from PhCl are approximately 100% already at T<400 degrees C, and T(50%) congruent with 260 degrees C, over 200 degrees C lower than with AC alone. There is no appreciable decrease of activity after having injected >20 mmol PhCl per gram of catalyst at partial conversion. PMID- 12547341 TI - Increased PCDD/F formation in the bottom ash from fires of CCA-treated wood. AB - Bottom ash that was the result of the combustion of chromated copper arsenate (CCA) treated wood under controlled fire conditions showed an increase of several orders of magnitude in the levels of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), compared to that of untreated timber. Wood that has been pressure treated with CCA contains copper (II), which is known to catalyse the so called de novo formation of PCDD/Fs. Comparable levels of PCDD/Fs would be expected in residual ash from burning CCA-treated wood in backyard fires, stoves and wood heaters, as a consequence of similar combustion conditions. PMID- 12547343 TI - The cytoskeleton of Giardia lamblia. AB - Giardia lamblia is a ubiquitous intestinal pathogen of mammals. Evolutionary studies have also defined it as a member of one of the earliest diverging eukaryotic lineages that we are able to cultivate and study in the laboratory. Despite early recognition of its striking structure resembling a half pear endowed with eight flagella and a unique ventral disk, a molecular understanding of the cytoskeleton of Giardia has been slow to emerge. Perhaps most importantly, although the association of Giardia with diarrhoeal disease has been known for several hundred years, little is known of the mechanism by which Giardia exacts such a toll on its host. What is clear, however, is that the flagella and disk are essential for parasite motility and attachment to host intestinal epithelial cells. Because peristaltic flow expels intestinal contents, attachment is necessary for parasites to remain in the small intestine and cause diarrhoea, underscoring the essential role of the cytoskeleton in virulence. This review presents current day knowledge of the cytoskeleton, focusing on its role in motility and attachment. As the advent of new molecular technologies in Giardia sets the stage for a renewed focus on the cytoskeleton and its role in Giardia virulence, we discuss future research directions in cytoskeletal function and regulation. PMID- 12547344 TI - Why are parasite contingency genes often associated with telomeres? AB - Contingency genes are common in pathogenic microbes and enable, through pre emptive mutational events, rapid, clonal switches in phenotype that are conducive to survival and proliferation in hosts. Antigenic variation, which is a highly successful survival strategy employed by eubacterial and eukaryotic pathogens, involves large repertoires of distinct contingency genes that are expressed differentially, enabling evasion of host acquired immunity. Most, but not all, antigenic variation systems make extensive use of subtelomeres. Study of model systems has shown that subtelomeres have unusual properties, including reversible silencing of genes mediated by proteins binding to the telomere, and engagement in ectopic recombination with other subtelomeres. There is a general theory that subtelomeric location confers a capacity for gene diversification through such recombination, although experimental evidence is that there is no increased mitotic recombination at such loci and that sequence homogenisation occurs. Possible benefits of subtelomeric location for pathogen contingency systems are reversible gene silencing, which could contribute to systems for gene switching and mutually exclusive expression, and ectopic recombination, leading to gene family diversification. We examine, in several antigenic variation systems, what possible benefits apply. PMID- 12547345 TI - Mucin-type O-glycosylation in Fasciola hepatica: characterisation of carcinoma associated Tn and sialyl-Tn antigens and evaluation of UDP-GalNAc:polypeptide N acetylgalactosaminyltransferase activity. AB - Simple mucin-type cancer-associated O-glycan structures, such as the Tn antigen (GalNAc-O-Ser/Thr), are expressed by certain helminth parasites. These antigens are involved in several types of receptor-ligand interactions, and they are potential targets for immunotherapy. The aim of this work was to study the initiation pathway of mucin-type O-glycosylation in Fasciola hepatica, performing a biochemical and immunohistochemical characterisation of Tn and sialyl-Tn antigens, and evaluating the ppGaNTase activity, which catalyses the first step in O-glycan biosynthesis. Using ELISA, both Tn and sialyl-Tn antigens were detected predominantly in the somatic and deoxycholate extracts. Immunofluorescence analysis revealed that Tn antigen is preferentially expressed in testis, while sialyl-Tn glycoproteins were more widely distributed, being present in parenchymal cells, basal membrane of the tegument, and apical surface of epithelial cells lining the caeca. On the basis of their electrophoretic mobility, Tn glycoproteins were resolved as six components of 10, 37, 76, 125, 170 and 205 kDa, and sialyl-Tn components showed an apparent molecular mass of 28 and 32 kDa, and two broad bands of 90-110 and 170-190 kDa. The observation that only the 76 kDa Tn-glycoprotein remained in the 0.6 N perchloric acid-soluble fraction suggests that it could be a good candidate for mucin characterisation in this parasite. The ppGaNTase activity showed its maximal activity at pH 7-7.5 and 37 degrees C, showing that Mn(2+) was the best divalent cation activator. Using a panel of nine synthetic peptides as acceptor substrates, we found that F. hepatica ppGaNTase was able to glycosylate both threonines and serines, the best substrates being the peptides derived from the tandem repeat region of human mucins (MUC2 and MUC6), and from Trypanosoma cruzi and Trypanosoma brucei glycoproteins. The results reported here constitute the first evidence on O glycosylation pathways in F. hepatica, and may help to identify new biological characteristics of this parasite as well as of the host-parasite relationship. PMID- 12547346 TI - Parasite altered micro-distribution of Gammarus pulex (Crustacea: Amphipoda). AB - In a river survey, Gammarus pulex amphipods both unparasitised and parasitised with the acanthocephalan Echinorhynchus truttae were distributed similarly with respect to flow regimen, tending to be more abundant in faster, shallower, riffle patches. However, there was a higher prevalence of parasitism in faster, shallower areas than in slower, deeper areas and abundance correlated with macrophyte coverage for unparasitised but not parasitised amphipods, indicating subtle differences in habitat usage. A laboratory 'patch' simulation indicated that parasitism influenced micro-distribution. There were higher proportions of unparasitised amphipods in/under stone substrates and within weed. In contrast, there were higher proportions of parasitised amphipods in the water column and at the water surface. As the experiment progressed, unparasitised but not parasitised amphipod habitat usage shifted from those micro-habitats above the substrate and in the water column to those in/under the substrates. Experiments also demonstrated that parasitised amphipods were more active and had a greater preference for illumination. Previous studies of the effects of acanthocephalan parasitism of amphipod hosts have focussed on how drift behaviour is altered, now we show that subtle differences in micro-habitat usage could translate to greatly increased vulnerability to fish predation. We discuss how aggregation of parasitised individuals within specific habitats could promote parasite transmission. PMID- 12547347 TI - Pomphorhynchus laevis (Palaeacanthocephala) in the intestine of chub (Leuciscus cephalus) as an indicator of metal pollution. AB - Chub experimentally infected with the acanthocephalan parasite Pomphorhynchus laevis were exposed to 0.01 mg l(-1) Pb(2+) over 5 weeks. Lead was rapidly accumulated in the intestinal worms reaching a steady-state concentration after 4 weeks. This concentration was significantly greater than in the host muscle, liver and intestine and more than 9,000 times higher than the exposure concentration. Lead accumulation in P. laevis increased at a higher exposure concentration but was not correlated with either parasite intensity or with pooled or individual worm weight. The highest lead concentrations were recorded in those specimen of P. laevis that were attached in the posterior intestine. Interestingly, parasitised chub accumulated less lead in their own tissues than uninfected conspecifics. A mechanism of lead uptake by P. laevis and the application of acanthocephalans as accumulation indicators of metal pollution is discussed. PMID- 12547348 TI - Successful vaccination of BALB/c mice against human hookworm (Necator americanus): the immunological phenotype of the protective response. AB - In this murine (BALB/c) model of necatoriasis, high levels of protection against challenge infection by Necator americanus larvae (n=300) were afforded by successive vaccinations at 14-day intervals, either subcutaneously or percutaneously, with gamma-irradiated N. americanus larvae (n=300). Percutaneous vaccination was significantly more effective than the subcutaneous route, with pulmonary larval burdens at 3 days post-infection being reduced by 97.8 vs. 89.3%, respectively, after three immunisations (P<0.05). No worms were recovered from the intestines of thrice vaccinated mice. Two percutaneous vaccinations also reduced worm burdens, by 57% in the lungs and 98% in the intestines; P<0.05. In vaccinated animals, lung pathology (mainly haemorrhage) following infection was greatly reduced compared with non-vaccinated animals. In vaccinated mice (but not in non-vaccinated mice) mast cells accumulated in the skin and were degranulated. RT-PCR analyses of mRNAs in the skin of vaccinated animals indicated increased expression of interleukin (IL)-4 relative to gamma-interferon (gamma-IFN). Lymphocytes from the axillary (skin-draining) lymph nodes of vaccinated mice, stimulated in vitro with concanavalin A, exhibited enhanced secretion of IL-4 protein and a higher IL-4/gamma-IFN protein ratio than lymphocytes from non vaccinated animals. In vaccinated mice, levels of IgG1 and IgG3 (directed against larval excretory/secretory products) were elevated for the most part compared with those in non-vaccinated animals. These data demonstrate the successful vaccination of BALB/c mice against human hookworm infection and suggest that a localised Th2 response may be important for conferring protection against necatoriasis. PMID- 12547349 TI - A single dose of recombinant Salmonella typhimurium induces specific humoral immune responses against heterologous Eimeria tenella antigens in chicken. AB - Salmonella typhimurium vaccine strains were used as antigen delivery system for oral immunisation of chickens against two antigens of the coccidian parasite Eimeria tenella. The cDNAs of the known E. tenella proteins, SO7 and TA4, were isolated from total RNA and subcloned into the expression vectors pQE30 and pTECH2. Subcutaneous immunisation of chickens with Escherichia coli-expressed SO7 and TA4 revealed that both proteins were immunogenic. Both cDNAs were subcloned into plasmids of the pTECH2 vector system, which allows them to be expressed as fusion proteins with the highly immunogenic fragment C of the tetanus toxin under control of the anaerobically inducible nirB promoter. Plasmids were introduced into the S. typhimurium vaccine strains SL3261, C5aroD and C5htrA. SDS-PAGE and Western blot analysis revealed expression of both fusion proteins in all strains under anaerobic culture conditions. Three-week-old white leghorn chickens were orally immunised with 10(9) CFU per animal. The stability of the recombinant bacteria was revealed by recovery of viable Salmonella containing the respective plasmids from the liver of the immunised chickens at day 3 after inoculation. Specific serum IgG antibodies against the SO7-or TA4-antigens were detectable by ELISA 2 weeks after oral immunisation and remained for at least 6 weeks, while specific IgA antibodies were restricted to the bile of the birds. All chickens produced serum IgG and IgA to S. typhimurium lipopolysaccharides. Our data show that a single oral inoculation with recombinant S. typhimurium SL3261, C5aroD and C5htrA can induce specific antibody responses to heterologous Eimeria antigens in chickens, suggesting that recombinant Salmonella are a suitable delivery system for vaccines against Eimeria infections. PMID- 12547350 TI - Organisation and sequence determination of glutamine-dependent carbamoyl phosphate synthetase II in Toxoplasma gondii. AB - Carbamoyl phosphate synthetase II encodes the first enzymic step of de novo pyrimidine biosynthesis. Carbamoyl phosphate synthetase II is essential for Toxoplasma gondii replication and virulence. In this study, we characterised the primary structure of a 28kb gene encoding Toxoplasma gondii carbamoyl phosphate synthetase II. The carbamoyl phosphate synthetase II gene was interrupted by 36 introns. The predicted protein encoded by the 37 carbamoyl phosphate synthetase II exons was a 1,687 amino acid polypeptide with an N-terminal glutamine amidotransferase domain fused with C-terminal carbamoyl phosphate synthetase domains. This bifunctional organisation of carbamoyl phosphate synthetase II is unique, so far, to protozoan parasites from the phylum Apicomplexa (Plasmodium, Babesia, Toxoplasma) or zoomastigina (Trypanosoma, Leishmania). Apicomplexan parasites possessed the largest carbamoyl phosphate synthetase II enzymes due to insertions in the glutamine amidotransferase and carbamoyl phosphate synthetase domains that were not present in the corresponding gene segments from bacteria, plants, fungi and mammals. The C-terminal allosteric regulatory domain, the carbamoyl phosphate synthetase linker domain and the oligomerisation domain were also distinct from the corresponding domains in other species. The novel C terminal regulatory domain may explain the lack of activation of Toxoplasma gondii carbamoyl phosphate synthetase II by the allosteric effector 5 phosphoribosyl 1-pyrophosphate. Toxoplasma gondii growth in vitro was markedly inhibited by the glutamine antagonist acivicin, an inhibitor of glutamine amidotransferase activity typically associated with carbamoyl phosphate synthetase II, guanosine monophosphate synthetase, or CTP synthetase. PMID- 12547351 TI - Molecular evidence for multiple Toxoplasma gondii infections in individual patients in England and Wales: public health implications. AB - We sought to determine the SAG2 genotypes of Toxoplasma gondii associated with cases of acute human toxoplasmosis in England and Wales. The samples examined were collected from a wide range of cases including congenital infections, AIDS and immunosuppressed patients and were derived from a number of different tissues. Parasite DNA was detected by PCR amplification without the need for prior template purification, and SAG2 genotype was determined by both restriction enzyme analysis and direct DNA sequencing of the PCR amplification products. Parasites of both SAG2 type I and type II genotypes were seen with approximately equal frequency amongst the samples examined. Neither of these genotypes was found to be more frequently associated with a particular clinical presentation or sample tissue. Unexpectedly, we found clear evidence of mixed (SAG2 type I+type II) infections in approximately the same number of samples as were seen to be associated with either type I or II alone. Our use of direct DNA sequencing rather than simple restriction analysis was essential for the detection of mixed infections since incomplete restriction digestion of samples containing a single parasite type was occasionally observed. It is possible that the presence of more than one type of parasite in single samples might be related to our recent demonstration that mixtures of SAG2 type I and type II parasites are present in a significant proportion of commercial meat preparations. Moreover, the presence of mixed infections in single patients might offer a direct molecular method of assessing risk factors for infection. PMID- 12547352 TI - Trabecular bone strain changes associated with subchondral stiffening of the proximal tibia. AB - Subchondral stiffening is a hallmark pathologic feature of osteoarthritis but its mechanical and temporal relationship to the initiation or the progression of osteoarthritis is not established. The mechanical effect of subchondral stiffening on the surrounding trabecular bone is poorly understood. This study employs a relatively new application of digital image correlation to measure strain in the trabecular region of the proximal medial tibia in normal specimens and in specimens with simulated subchondral bone stiffening. Coronal sections from eight normal human cadaveric proximal tibiae were loaded in static compression and high resolution contact radiographs were made. Repeat contact radiographs were collected after the subchondral bone near the jointline was stiffened using polymethylmethacrylate. Digital images, made from loaded and unloaded contact radiographs, were compared using in-house software to measure trabecular displacement and calculate trabecular bone strain. Overall strain was higher in the stiffened specimens suggesting experimental artifiact significantly affected our results. Consistent increases in median maximum shear strain, median maximum principal strain, median minimum principal strain, and peak shear strain were measured near the inner and outer edges of the stiffened segment. Our experiment provides direct experimental measurement of increases in trabecular bone strain caused by subchondral stiffening, however, the clinical and biologic importance of strain increases is unknown. PMID- 12547353 TI - Determination of constitutive equations for human arteries from clinical data. AB - Stress-strain analyses of vessel walls require appropriate constitutive equations. Determination of constitutive equations is based on experimental data of (i) diameter and length of a vessel segment subject to internal pressure and external axial force, and (ii) the load-free reference geometry. Typical clinical data, however, provide only pressure-diameter relations in the diastolic-systolic pressure range. In order to overcome this problem, an approach is proposed allowing the determination of constitutive equations from clinical data by means of reasonable assumptions regarding in situ configurations and stress states of arterial walls. The approach is based on a two-dimensional Fung-type stored energy function capturing the characteristic nonlinear and anisotropic responses of arteries. Examples concerning human aortas from a normotensive and a hypertensive subject illustrate the potential of the approach. PMID- 12547354 TI - An in vitro investigation of the acetabular labral seal in hip joint mechanics. AB - Labrum pathology may contribute to early joint degeneration through the alteration of load transfer between, and the stresses within, the cartilage layers of the hip. We hypothesize that the labrum seals the hip joint, creating a hydrostatic fluid pressure in the intra-articular space, and limiting the rate of cartilage layer consolidation. The overall cartilage creep consolidation of six human hip joints was measured during the application of a constant load of 0.75 times bodyweight, or a cyclic sinusoidal load of 0.75+/-0.25 times bodyweight, before and after total labrum resection. The fluid pressure within the acetabular was measured. Following labrum resection, the initial consolidation rate was 22% greater (p=0.02) and the final consolidation displacement was 21% greater (p=0.02). There was no significant difference in the final consolidation rate. Loading type (constant vs. cyclic) had no significant effect on the measured consolidation behaviour. Fluid pressurisation was observed in three of the six hips. The average pressures measured were: for constant loading, 541+/-61kPa in the intact joint and 216+/-165kPa following labrum resection, for cyclic loading, 550+/-56kPa in the intact joint and 195+/-145kPa following labrum resection. The trends observed in this experiment support the predictions of previous finite element analyses. Hydrostatic fluid pressurisation within the intra-articular space is greater with the labrum than without, which may enhance joint lubrication. Cartilage consolidation is quicker without the labrum than with, as the labrum adds an extra resistance to the flow path for interstitial fluid expression. However, both sealing mechanisms are dependent on the fit of the labrum against the femoral head. PMID- 12547355 TI - Deformation behaviour and damage accumulation of cortical bone specimens from the equine tibia under cyclic loading. AB - Despite its clinical importance, the fatigue behaviour of cortical bone has not been examined as widely as its static behaviour. In the present study, specimens from the tibiae of horses have been subjected to load-controlled single step tests. The cyclic deformation behaviour was described by the development of stress-strain hysteresis parameters over the lifetime. The fatigue behaviour of bone is characterised by cyclic softening which is most distinctive towards the end of the lifetime. The microstructural damage accumulated during cyclic loading results in a loss of stiffness, asymmetrical deformation of the bone in tension and compression in cyclic creep. As shown by light and scanning electron microscopy, microcrack formation and growth is the main damage mechanism. The crack growth behaviour is strongly influenced by the microstructure, the stress components and the absolute value of the local stresses. Lower local stresses and/or compressive mean stresses lead to a dominant influence of the shear stress components with shear failure at inner interfaces. With increasing crack length, that is, higher local stress amplitudes, or tensile mean stresses, the microstructure is more and more ignored and failure occurs primarily under the influence of the normal stress components. This can be clearly seen on the fracture and specimen surfaces. PMID- 12547356 TI - Equine cortical bone exhibits rising R-curve fracture mechanics. AB - Previous studies of the fracture properties of cortical bone have suggested that the fracture toughness increases with crack length, which is indicative of rising R-curve behavior. Based on this indirect evidence and the similarity of bone to ceramic matrix composites, we hypothesized that bone would exhibit rising R-curve behavior in the transverse orientation and that the characteristics of the R curves would be regionally dependent within the cortex due to variations in bone microstructure and toughening mechanisms. To test these hypotheses, we conducted R-curve experiments on specimens from equine third metacarpal bones using standard fracture mechanics testing methods. Compact type specimens from the dorsal and lateral regions in the middle of the diaphysis were oriented for crack propagation transverse to the longitudinal axis of the bone. The test results demonstrate that equine cortical bone exhibits rising R-curve behavior during transverse crack propagation as hypothesized. Statistical analyses of the crack growth initiation toughness, K0, the peak toughness, Kpeak, and the crack extension at peak toughness, deltaa, revealed significant regional differences in these characteristics. Specifically, the lateral cortex displayed higher crack growth initiation and peak toughnesses. The dorsal cortex exhibited greater crack extension at the peak of crack growth resistance. Scanning electron microscopy revealed osteon pullout on fracture surfaces from the dorsal cortex and but not in the lateral cortex. Taken together, the significant differences in R-curves and the SEM fractography indicate that the fracture mechanisms acting in equine cortical bone are regionally dependent. PMID- 12547357 TI - Reflex muscle contractions can be elicited by valgus positional perturbations of the human knee. AB - Experimental evidence on the reflex responses of thigh muscles to valgus mechanical perturbations at the human knee are presented. Random step positional deflections, ranging from 5 degrees to 12 degrees at 60 degrees /s, were applied to the fully extended knees of seven healthy subjects. Subjects were instructed to maintain a constant background co-activation ( approximately 2-11% MVC) of the quadriceps and hamstring muscles prior to and during the mechanical stimulus. We found that the reflex response to sustained valgus joint deflection in the vasti muscles had longer onset latencies (range: 83-92ms) than did the stretch reflex in the same muscles (latencies: 29-31ms). This reflex EMG response consisted typically of a peak followed by sustained muscle activity throughout the step perturbation. The sustained EMG activity was dependent on the amplitude of the perturbing stimulus, but in a nonlinear manner. The long latency of the valgus response suggests that the reflex originates in nonmuscular sensory pathways, potentially from mechanoreceptors lying in periarticular tissues such as joint ligaments and capsule. Analysis of the spatial distribution of reflex responses showed an asymmetrical pattern with preferential activation of medial vs. lateral muscles of the knee. We assess whether these asymmetric reflex contractions could promote joint stability, either by inducing generalized joint stiffening, or by preferential activation of those muscles that are best suited to resist induced ligament strain. PMID- 12547358 TI - Hill muscle model errors during movement are greatest within the physiologically relevant range of motor unit firing rates. AB - This study evaluated the accuracy of Hill-type muscle models during movement. Hill-type models are ubiquitous in biomechanical simulations. They are attractive because of their computational simplicity and close relation to commonly measured experimental variables, but there have been surprisingly few experimental validations of these models during functionally relevant conditions. Our hypothesis was that model errors during movement are largest at the low motor unit firing rates most relevant to normal movement conditions. This hypothesis was evaluated in the cat soleus muscle activated either by electrical stimulation at physiological rates or via the crossed-extension reflex (CXR) thereby obtaining normal patterns of motor unit recruitment and rate modulation. These activation paradigms were applied during continuous movements approximately matched to locomotor length changes. The resulting muscle force was modeled using a common Hill model incorporating independent activation, tetanic length-tension and tetanic force-velocity properties. Errors for this model were greatest for stimulation rates between approximately 10-20Hz. Errors were especially large for muscles activated via the CXR, where most motor units appear to fire within this range. For large muscle excursions, such as those seen during normal locomotion, the errors for naturally activated muscle typically exceeded 50%, supporting our hypothesis and indicating that the Hill model is not appropriate for these conditions. Subsequent analysis suggested that model errors were due to the common Hill model's inability to account for the coupling between muscle activation and force-velocity properties that is most prevalent at the low motor unit firing rates relevant to normal activation. PMID- 12547359 TI - Bone mineral density correlates with fracture load in experimental side impacts of the pelvis. AB - Pelvic fractures resulting from automotive side impacts are associated with high mortality and morbidity, as well as substantial economic costs. Previous experimental studies have produced varying results regarding the tolerance of the pelvis to lateral force and compression. While bone mineral density (BMD) has been shown to correlate with fracture loads in the proximal femur, no such correlation has been established for the pelvis. Presently, we studied the relationships between total hip BMD and impact response parameters in lateral impacts of twelve isolated human pelves. The results indicated that total hip BMD significantly correlated with fracture force, Fmax, and maximum ring compression, Cmax, of the fractured pelves. These findings are evidence that BMD may be useful in assessing the risk of pelvic fracture in automotive side impacts. Poor correlation was observed between total hip BMD and maximum viscous response, (VC)max, energy at fracture, Epeak, and time to fracture, tpeak. Mean Fmax and calculated tolerances for Cmax and (VC)max were lower than those established in previous studies using full cadavers, likely a result of our removal of soft tissues from the pelves prior to impact. PMID- 12547360 TI - Fracture-toughening mechanisms responsible for differences in work to fracture of hydrated and dehydrated dentine. AB - This study investigates the nature of deformation and differences in the mechanisms of fracture and properties of dentine where there has been a loss of moisture, as may occur with removal of the pulp in the endodontic treatment of teeth. Controlled fracture toughness testing was conducted on bovine teeth to determine the influence of hydration on the work of fracture of dentine. Significant differences (p<0.01) were observed between the fracture toughness of hydrated (554+/-27.7J/m2) and dehydrated (113+/-17.8J/m2) dentine. Observations of the crack tip region during crack extension revealed extensive ligament formation occurred behind the crack tip. These ligaments provide considerable stability to the crack by significantly increasing the work of fracture, thereby acting as a fracture-toughening mechanism. Micro-cracking, reported as a fracture toughening mechanism in bone, is also clearly seen. A zone of in-elastic deformation may occur as hydrated specimens revealed upon crack extension, a region about the tip that appeared to suck water into the structure and to exude water behind the crack tip. In dehydrated dentine, no in-elastic zone was observed. Micro-cracking is present though the cracks are smaller, straighter and with less opening than hydrated dentine. Only limited ligament formation just behind the crack tip was observed. These differences resulted in a significantly lower work of fracture with unstable brittle fracture characteristics. Based on these results, several fracture-toughening mechanisms were identified in dentine, with micro-cracking not considered the most important. These findings may be relevant for bone, a similar mineralised hydrated tissue. PMID- 12547361 TI - The push force pattern in manual wheelchair propulsion as a balance between cost and effect. AB - We investigate the hypothesis that the direction of the propulsion force in manual wheelchair propulsion can be interpreted as a result of the balance between the mechanical task requirements and the driver's biomechanical possibilities. We quantify the balance at the joint level in the form of an effect-cost criterion, from which we predict the force direction that results in an optimal compromise. Kinematic and dynamic data were collected from nine habitual wheelchair users driving at four velocities (0.83, 1.11, 1.39, 1.67 m/s) and three external power levels (10, 20, 30 W). Experimental data and predictions are in good agreement in the middle and final part of the push; the effect-cost value in this region approximates the achievable maximum. Early in the push the effect-cost criterion predicts an upwards propulsion force whereas the experimental force is downwards, the difference probably being mainly attributable to the force generation dynamics of the muscles. As a result of the geometric features of large-rim manual wheelchairs, the mechanically required and biomechanically preferred force directions are not in accordance during a substantial part of the push, making even the best compromise a poor one. This may contribute to the low mechanical efficiency of manual wheelchair propulsion and the high incidence of shoulder complaints. PMID- 12547362 TI - Optimum ratio of upper to lower limb lengths in hand-carrying of a load under the assumption of frequency coordination. AB - The ratio of the upper to lower limb lengths [or the intermembral index (IMI)] in the earliest human ancestors is closer to that of the living chimpanzees than to our own, although the former show undoubted adaptations to bipedality. What biomechanical factors could then have led to the phenomenon of genus Homo? This paper proposes and evaluates a relationship between IMI and hand-carrying. Assuming that coordination of limb swing frequencies of the upper and lower limbs would be the subject of positive selection, a mathematical expression was derived and can in part explain the changes in IMI. We found that AL-288-1 [3.6 million years old (MY)], the most complete skeleton of the early hominid Australopithecus afarensis, could only have carried loads equivalent to 15-50% of the upper limb weight while maintaining swing symmetry, but KNM WT-15000, Homo ergaster (1.8MY) and modern humans could both carry loads 3 times heavier than the upper limb while maintaining swing symmetry. The carrying ability of chimpanzees would be inferior to that of AL-288-1. The IMI of modern humans, at 68-70, is the smallest, and is optimal for hand-carrying under our criteria. Under reduced selection pressure for hand-carrying, but unreduced selection for mechanical effectiveness, we might expect humans to evolve a longer upper limb, to improve swing symmetry when unloaded. PMID- 12547363 TI - A telemetry-based velocometer to measure wheelchair velocity. AB - The purpose of this paper is to present a telemetry-based velocometer that has the ability to measure wheelchair velocity. Five studies are described which provide measurements of the validity, dynamic response, reliability and resistance of the velocometer. VALIDITY: a linear relationship was found when velocity calculated from the velocometer was plotted against three test velocities. The average root mean square deviation (ARMSD) was used to compare velocity calculated from the velocometer with velocity calculated by manual digitising at 200Hz. The ARMSD calculated for each test speed from three trials were 0.06+/-0.01, 0.27+/-0.05 and 0.48+/-0.16 ms(-1) at 1, 5 and 9 ms(-1), respectively. Dynamic response: expressed as a percentage of the average mean trial velocity, the ARMSD for the five acceleration and five deceleration trials were 6.5+/-1.8% and 6.9+/-1.2%, respectively. Reliability was assessed from a comparison between mean trial velocity calculated from velocometer output and the speed of the motor used to spin the wheels. Expressed as a percentage of the mean trial velocity, the mean+/-SD of the differences were 0.00+/-0.17%, for the ten disc wheel trials and 0.00+/-0.41%, for the ten spoke wheel trials. Velocometer resistance calculated as a factor of the mechanical resistance of the wheelchair rear wheel spinning in air were -0.50 and -0.91 N, for the disc and spoke wheel, respectively. Velocometer resistance calculated as a factor of the total mechanical resistance of the wheelchair-wheelchair user system were -1.37 and 1.82 N, for the disc and spoke wheel, respectively. PMID- 12547364 TI - Improved RSA accuracy with DLT and balanced calibration marker distributions with an assessment of initial-calibration. AB - Roentgen stereophotogrammetric analysis (RSA) has been used for over 25 years for accurate micromotion measurement in a wide variety of orthopaedic applications. This study investigated two possible improvements to the method. First, direct linear transformation (DLT) was compared to the traditional RSA reconstruction algorithm. The two methods were considered with respect to standard extrapolation and interpolation calibration cages. Matlab simulations showed that reconstruction accuracy was greatly improved (>60%) by combining DLT with an even distribution of enclosing calibration markers. Second, a benchtop study using phantoms translated at 0.0254-mm intervals showed initial-calibration, followed by removal of the interpolation cage for subsequent exposures, was potentially twice as accurate as self-calibration with an extrapolation cage. These results showed optimizations for the application of RSA when unobstructed space is required. PMID- 12547365 TI - The strength-dexterity test as a measure of dynamic pinch performance. AB - We have developed a method to quantify the dynamic interaction between fingertip force magnitude (strength) and directional control (dexterity) during pinch with a novel strength-dexterity (S-D) test based on the principle of buckling of compression springs. The test consists of asking participants to use key and opposition pinch to attempt to fully compress springs, in random order, with a wide range of combinations of strength and dexterity requirements. The minimum force required to fully compress the spring and the propensity of the spring to buckle define the strength and dexterity requirements, respectively. The S-D score for each pinch style was the sum of the strength values of all springs successfully compressed fully. We tested 3 participant groups: 18 unimpaired young adults (40yr), and 14 adults diagnosed with carpo-metacarpal osteoarthritis (CMC OA) (>or = 36yr). We investigated the repeatability of the S-D test with 74 springs by testing 14 young adults twice on different days. The per-spring repeatability across subjects was >or = 94%. A minimum performance score for young adults was found as they all could compress a subset of 39 springs. Using this subset of springs, we compared the ability of the S-D score vs. maximal pinch force values to distinguish unimpaired hands from those with CMC OA of the thumb. The score for this 39-spring S-D test distinguished between CMC OA and asymptomatic older adults, whereas pinch meter readings did not (p<0.05). We conclude that the S-D test is repeatable and applicable to clinical research. We propose including the S-D test in studies aiming to quantify impairment and compare treatment outcomes in orthopaedic and neurological afflictions that degrade dynamic manipulation. PMID- 12547366 TI - Limitations in the use and interpretation of continuous relative phase. AB - Continuous relative phase (CRP), a variable used to quantify intersegmental coordination, is difficult to interpret if care is not taken regarding the assumptions and limitations of the measure. Specifically, CRP is often interpreted as a higher resolution form of discrete relative phase (DRP). DRP, however, yields information regarding the relative dispersion of events in oscillatory signals while CRP describes their relationship in a higher order phase-plane domain. In this paper we address issues surrounding the calculation of CRP and suggest a new interpretation based on the aforementioned methodological issues. Through the use of test signals, with known properties, it was found that the CRP information will be arbitrary if no normalization procedures are used to account for frequency differences in the component oscillators. In addition, signals with non-sinusoidal trajectories will produce patterns in CRP that are not equivalent to discrete relative phase (DRP) measures. The implications of these issues are discussed. PMID- 12547367 TI - Mandibular kinematics represented by a non-orthogonal floating axis joint coordinate system. AB - There are many methods used to represent joint kinematics (e.g., roll, pitch, and yaw angles; instantaneous center of rotation; kinematic center; helical axis). Often in biomechanics internal landmarks are inferred from external landmarks. This study represents mandibular kinematics using a non-orthogonal floating axis joint coordinate system based on 3-D geometric models with parameters that are "clinician friendly" and mathematically rigorous. Kinematics data for two controls were acquired from passive fiducial markers attached to a custom dental clutch. The geometric models were constructed from MRI data. The superior point along the arc of the long axis of the condyle was used to define the coordinate axes. The kinematic data and geometric models were registered through fiducial markers visible during both protocols. The mean absolute maxima across the subjects for sagittal rotation, coronal rotation, axial rotation, medial-lateral translation, anterior-posterior translation, and inferior-superior translation were 34.10 degrees, 1.82 degrees, 1.14 degrees, 2.31, 21.07, and 6.95 mm, respectively. All the parameters, except for one subject's axial rotation, were reproducible across two motion recording sessions. There was a linear correlation between sagittal rotation and translation, the dominant motion plane, with approximately 1.5 degrees of rotation per millimeter of translation. The novel approach of combining the floating axis system with geometric models succinctly described mandibular kinematics with reproducible and clinician friendly parameters. PMID- 12547368 TI - Performance characteristics of a pressure microsensor. AB - A new fiber optic microsensor has been developed for pressure measurement. The pressure microsensor is extremely small (360 micro m). Performance characteristics of the microsensor were evaluated over a pressure range from 0 to 250mmHg. Five sensors were compared to a National Institute of Standards and Technology traceable reference pressure. The microsensor had an accuracy, repeatability, and linearity better than 2% full-scale output (FSO) and a hysteresis of 4.5% FSO. PMID- 12547369 TI - Concept and development of an orthotropic FE model of the proximal femur. AB - PURPOSE: In contrast to many isotropic finite-element (FE) models of the femur in literature, it was the object of our study to develop an orthotropic FE "model femur" to realistically simulate three-dimensional bone remodelling. METHODS: The three-dimensional geometry of the proximal femur was reconstructed by CT scans of a pair of cadaveric femurs at equal distances of 2mm. These three-dimensional CT models were implemented into an FE simulation tool. Well-known "density determined" bony material properties (Young's modulus; Poisson's ratio; ultimate strength in pressure, tension and torsion; shear modulus) were assigned to each FE of the same "CT-density-characterized" volumetric group. In order to fix the principal directions of stiffness in FE areas with the same "density characterization", the cadaveric femurs were cut in 2mm slices in frontal (left femur) and sagittal plane (right femur). Each femoral slice was scanned into a computer-based image processing system. On these images, the principal directions of stiffness of cancellous and cortical bone were determined manually using the orientation of the trabecular structures and the Haversian system. Finally, these geometric data were matched with the "CT-density characterized" three-dimensional femur model. In addition, the time and density-dependent adaptive behaviour of bone remodelling was taken into account by implementation of Carter's criterion. RESULTS: In the constructed "model femur", each FE is characterized by the principal directions of the stiffness and the "CT-density-determined" material properties of cortical and cancellous bone. Thus, on the basis of anatomic data a three-dimensional FE simulation reference model of the proximal femur was realized considering orthotropic conditions of bone behaviour. CONCLUSIONS: With the orthotropic "model femur", the fundamental basis has been formed to realize realistic simulations of the dynamical processes of bone remodelling under different loading conditions or operative procedures (osteotomies, total hip replacements, etc). PMID- 12547370 TI - A new testing device for measuring gliding resistance and work of flexion in a digit. AB - In order to move the finger the tendon force must overcome the gliding resistance of the tendon as well as the forces to move the joints, finger inertias, and external load. These sources, combined, make up the work of flexion (WOF) which has been experimentally used to evaluate the finger function. In this study, we have designed a new device, which can measure the forces at the proximal and distal end of the tendon during finger flexion, so that gliding resistance can be isolated from the WOF. Two index fingers from a pair of human cadaver hands were used for testing this device. Preliminary data showed that internal resistance occupied about 10% of WOF with an intact tendon. However, after tendon repair, the gliding resistance increased 31% of WOF for a modified Kessler repair and 50% of WOF for a Becker repair compared to intact tendon. We simulated joint stiffness by injection of saline solution into the proximal interphalangeal joint. This increased the overall WOF but not the gliding resistance. We believe that this testing device provides a useful tool to evaluate finger function after tendon repair in an experimental model. PMID- 12547371 TI - ISB recommendation on definition of joint coordinate systems for the reporting of human joint motion-part I: ankle, hip and spine. PMID- 12547372 TI - Comments on "Mechanical properties of single hyaluronan molecules" (J Biomech. 35(4) (2002) 527-531). PMID- 12547375 TI - Spontaneous decay of compulsive urges: the case of covert compulsions. AB - It is well established that many patients with obsessive-compulsive disorder have covert, or internal, compulsions. Empirical studies of this phenomenon, however, are limited. The present study followed the paradigm developed by Rachman and his colleagues for the study of overt compulsions. Patients with urges to carry out covert compulsions underwent an experimental procedure in which their compulsive urges were provoked, followed by a period of response prevention. The strength of the compulsive urges and associated discomfort were monitored. There was marked, and relatively rapid, spontaneous decay of both the compulsive urges and the discomfort. Implications of these results are discussed. PMID- 12547376 TI - Psychophysiologic effects of applied tension on the emotional fainting response to blood and injury. AB - OBJECTIVE: The present study was designed to investigate the psychophysiologic effects of "Applied Tension" (AT) on the emotional fainting response to blood and injury in a controlled experiment. METHOD: Twenty-two persons reporting to generally feel faint or to have fainted at the sight of blood or injury and 22 participants classified as Non-Fainters were randomly allocated to a treatment or control condition. Psychophysiologic responses were continuously monitored while individuals watched a video depicting open-heart surgery and a control film. Prior to the surgery film, participants in the treatment condition were instructed in the use of AT. RESULTS: All participants classified as Fainters showed a diphasic response pattern while watching the surgery film. This response, however, was significantly attenuated in Fainters in the treatment condition. CONCLUSIONS: These results suggest that AT provides an effective treatment strategy for the prevention of fainting responses in persons with a fear of blood and injury. PMID- 12547377 TI - Peritraumatic dissociation and PTSD severity: do event-related fears about death and control mediate their relation? AB - Relations among peritraumatic dissociation, PTSD severity, event-related fear (i.e. fear experienced during traumatic event) about death, and event-related fear about losing control were examined in the current study. Particular emphasis was placed on testing whether or not fears about death and losing control mediate the relation between peritraumatic dissociation and PTSD severity in a sample of 146 nontreatment-seeking university women. Results indicated that event-related fears about death and losing control accounted for the relation between peritraumatic dissociation and PTSD severity; that is, the effect of peritraumatic dissociation on PTSD severity was eliminated after controlling for these fears. Speculations about findings are discussed. PMID- 12547378 TI - Dissociation and memory fragmentation: experimental effects on meta-memory but not on actual memory performance. AB - The relation between state dissociation and fragmentary memory was investigated by assessing both actual memory performance and meta-memory. From a sample of 330 normal subjects, two subsamples were selected on basis of trait dissociation, as measured by the Dissociative Experience Scale. Twenty subjects scoring above 30 and 20 subjects scoring below 10 were selected. Subjects watched an extremely aversive film, after which state dissociation was measured by the Peri-traumatic Dissociative Experience Scale. Four hours later memory fragmentation was assessed in two ways. Actual fragmentation was measured by a sequential memory task, and perceived fragmentation (meta-memory) was measured using a visual analogue scale. Subjects who tended to dissociate during the film judged their recollections of the film as more fragmentary. Although this finding is in line with clinical reports given by trauma victims, it was not sustained by objective evidence. That is, no effect was observed of state dissociation on the sequential memory task. The present findings suggest that the claim that dissociation induces memory fragmentation may have to be confined to meta-memory. Implications of this divergence between actual memory and meta-memory are discussed. PMID- 12547379 TI - Hoarding behaviors in a large college sample. AB - Hoarding behaviors occur in many clinical syndromes but are most commonly linked to obsessive compulsive disorder. Surprisingly little empirical work has examined the nature of hoarding behaviors despite their association with significant distress and impairment. The current study examined hoarding in 563 unselected college students. Using principal components analysis, we identified four domains of hoarding behaviors as measured by the 26-item Saving Inventory-Revised: Difficulty Discarding, Acquisition Problems, Clutter, and Interference/Distress. All four domains and total hoarding behaviors were strongly related to hoarding beliefs and to obsessive compulsive (OC) symptoms. Hoarding behaviors were most strongly correlated with subscales of an OC disorder (OCD) measure assessing hoarding and obsessions and least strongly correlated with the washing subscale. Hoarding behaviors also showed significant, but more modest, correlations with social anxiety, depressive symptoms, and worry. However, worry was not found to contribute unique variance to the prediction of hoarding behaviors. Of greatest note, hoarding behaviors showed a surprisingly strong relationship with anxiety sensitivity, similar in magnitude to the relationship between hoarding and OCD symptoms. Results are interpreted and lines of future research are proposed, with particular emphasis on further elucidating the relationship between hoarding behaviors and anxiety sensitivity. PMID- 12547380 TI - Fear of the beast: a prospective study on the effects of negative information on childhood fear. AB - The current study examined the effects of negative information on the enhancement of childhood fear. A large group of normal primary school children aged between 4 and 12 years (N=285) received either negative or positive information about an unknown, doglike animal, called 'the beast'. Children's fears were assessed at three points in time: before, directly after, and one week after the information about the beast was provided (i.e., pre-, post- and follow-up assessment). Results showed that type of information changed children's fear of the beast in the predicted direction with negative information increasing fear levels and positive information decreasing fear levels. This was not only the case directly after the experimental manipulation but also at one week follow-up. Furthermore, fear of the beast appeared to generalize, that is, children who became more fearful of the beast after receiving negative information, also became more apprehensive of other dogs and predators. PMID- 12547381 TI - Shyness: relationship to social phobia and other psychiatric disorders. AB - The relationship between shyness, social phobia and other psychiatric disorders was examined. The prevalence of social phobia was significantly higher among shy persons (18%) compared with non-shy persons (3%). However, the majority of shy individuals (82%) were not socially phobic. A significant and positive correlation was found between the severity of shyness and the presence of social phobia, but the data suggest that social phobia is not merely severe shyness. Social phobia was also positively and moderately correlated with introversion and neuroticism. Thus, shy persons with social phobia were shyer, more introverted, and more neurotic than other shy people, but none of these factors was sufficient to distinguish shy persons with social phobia from those without social phobia. The proportion of the shy group with psychiatric diagnoses other than social phobia was significantly higher than among the non-shy group, indicating that various diagnostic categories are prominent among the shy. The results are discussed in terms of the overlap in shyness and social phobia and the relationship of shyness to other psychiatric diagnoses and personality dimensions. PMID- 12547382 TI - Pain anxiety among chronic pain patients: specific phobia or manifestation of anxiety sensitivity? AB - Rather than viewing anxiety among chronic pain patients as simply a component of negative affectivity, investigators have developed a model of "pain anxiety" in which patients develop fear and avoidance of activity linked to pain. We examined whether pain anxiety can be conceptualized as a specific phobia, or whether evidence supported the notion that pain anxiety is better understood as a manifestation of anxiety sensitivity in the context of chronic pain. Chronic musculoskeletal pain patients (N=70) underwent cold pressor and mental arithmetic tasks while cardiovascular, self-report, and behavior indexes were recorded. They completed measures of pain anxiety, anxiety sensitivity, fear of negative evaluation, depression and trait anxiety. Correlation analyses showed pain anxiety was related to pain-relevant responses during cold pressor, but it was also related to evaluation-relevant responses during cold pressor, and to pain- and evaluation-relevant responses (including subtraction accuracy) during mental arithmetic. Regression analyses showed that almost all effects of pain anxiety on task responses were accounted for by anxiety sensitivity. Fear of negative evaluation, in contrast, correlated only with evaluation-relevant responses, and mostly during mental arithmetic. These effects remained significant when depression, trait anxiety, or anxiety sensitivity were statistically controlled. Pain anxiety may be an expression of anxiety sensitivity rather than a circumscribed phobia; a distinction that could profitably guide treatment strategies. PMID- 12547383 TI - Behavioral activity associated with onset in chronic tic and habit disorder. AB - Seventy-six people (aged 18-62 years) diagnosed with either a chronic tic disorder or a habit disorder, entering a treatment study, kept a baseline daily diary for at least ten days, noting tic frequency and activity at time of onset. Together with an evaluator, participants completed a form ranking three high-risk activities where the tic or habit was likely to appear, and three low-risk activities where the tic was absent or barely present. Subjective appraisals distinguishing the two types of activities were also elicited and their relevance to the tic was measured on a seven-point scale using an adaptation of Kelly's repertory grid technique. Overall, the most frequent high-risk and low-risk activities were, respectively, passive attendance and physical activity. There were, however, significant differences in types of high-risk activities amongst the tic and habit disorders. Conversely, appraisals of the high-risk activities seemed to center on negative evaluations of tenseness, boredom, dissatisfaction, and disinterest. PMID- 12547384 TI - A comparison of metacognitions in patients with hallucinations, delusions, panic disorder, and non-patient controls. AB - This study tested the hypothesis that metacognitions are a general vulnerability factor for psychological disorder. It was predicted that patients with psychosis (hallucinations or delusions), and patients with panic disorder would score higher than non-patients on measures of metacognition. Moreover, it was hypothesised that patients showing most dysregulation of thinking (voice-hearers) would endorse significantly higher metacognition scores than individuals in the other groups. The Meta-Cognitions Questionnaire (MCQ: ) was administered to patients who met DSM-IV criteria for schizophrenia spectrum disorders with auditory hallucinations, patients who met DSM-IV criteria for schizophrenia spectrum disorders with persecutory delusions, patients who met DSM-IV criteria for panic disorder and non-patients. The results showed that psychotic patients who experience auditory hallucinations tended to exhibit higher levels of dysfunctional metacognitive beliefs than other patient groups, scoring significantly higher than at least two of the three control groups on positive beliefs about worry, negative beliefs about uncontrollability and danger, cognitive confidence and negative beliefs including superstition, punishment and responsibility. It was also found that the metacognitive beliefs of patients with persecutory delusions and panic patients were often similar to each other, and elevated in comparison to non-patients, suggesting that such beliefs are generic vulnerability factors. The theoretical and clinical implications of these findings are discussed. PMID- 12547388 TI - Induction of genome instability by DNA damage in Saccharomyces cerevisiae. AB - The accumulation of gross chromosomal rearrangements (GCRs) is a characteristic of many types of cancer cells, although it is unclear what defects cause these rearrangements and how the different types of GCRs observed are formed. In the present study, we have used a Saccharomyces cerevisiae system for measuring GCRs to analyze the ability of a variety of DNA damaging agents to induce GCRs. The two most potent inducers of GCRs observed were methyl methane sulfonate (MMS) and HO-endonuclease-induced double strand breaks (DSBs). Bleomycin, camptothecan and gamma-irradiation induced intermediate levels of GCRs and cisplatin induced very low levels of GCRs whereas N-methyl-NPRIME;-nitro-N-nitrosoguanidine (MNNG) and ethyl methane sulfonate (EMS) primarily induced base substitution mutations. MMS treatment primarily induced rearrangements in which the end of a chromosome was deleted and a new telomere was added (telomere additions) and also induced translocations. Consistent with this GCR spectrum, the formation of MMS-induced GCRs was primarily dependent on telomere maintenance functions and were completely eliminated in mutants that were defective for both telomere maintenance functions and non-homologous end joining (NHEJ). In contrast, HO endonuclease DSBs induced mostly translocations and interstitial deletions whereas few telomere additions were observed. Genetic analysis indicated that HO DSB-induced GCRs were suppressed by a number of pathways including the DNA damage checkpoints, DSB repair pathways and NHEJ. PMID- 12547389 TI - Specific stimulation of human apurinic/apyrimidinic endonuclease by heat shock protein 70. AB - We previously demonstrated the stimulation of human apurinic/apyrimidinic endonuclease 1 (HAP1) by heat shock protein 70 (HSP70). In this work, we further defined the functional interaction between these proteins. Digestion of HSP70 by trypsin released 48 and 43 kDa amino terminal fragments that retained the ability to stimulate HAP1. In agreement with this result, an HSP70 N-terminal deletion mutant protein containing amino acids 1-385 was comparable to the full-length protein in its ability to enhance HAP1 activity. HSP70 mutants containing carboxy terminal amino acids 386-640 stimulated HAP1 only slightly, as did unrelated proteins. These results implicate the amino terminal portion of HSP70 in stimulating the activity of HAP1. PMID- 12547390 TI - recX, a new SOS gene that is co-transcribed with the recA gene in Escherichia coli. AB - recX is a small open reading frame located downstream of recA that is conserved in many bacteria. In Escherichia coli, the recX gene (also named oraA) is a 501 bp open reading frame that encodes a predicted basic protein. Transcriptional analysis by Northern blots showed that in E. coli the recX gene is SOS-regulated. Primer extension data and transcriptional fusions indicate that recX transcription is down regulated with respect to recA by an intrinsic transcription terminator that is located between the recA and recX coding sequences. Despite the presence of this terminator, a recA-recX message resulting from transcriptional readthrough is detected at a level of 5-10% of the recA message. In addition, transcriptional/translational fusion experiments show that recX expression is further down regulated at the translational level reaching an estimated protein level about 500-fold lower than RecA. Strains in which the recX gene was disrupted were constructed by insertion of an antibiotic resistance cassette. The survival after UV irradiation, the spontaneous and UV-induced mutation rates were not significantly different in these recX strains compared to the corresponding wild type strain. Overexpression of RecA was shown to be lethal in a recX deletion strain in Pseudomonas aeruginosa [J. Bacteriol. 175 (1993) 2451], Mycobacterium tuberculosis [Mol. Microbiol. 30 (1998) 525] and Streptomyces lividans [J. Bacteriol. 182 (2000) 4005] suggesting that the recX gene may act as a regulator of recA. In contrast in E. coli, in a recX deletion strain, RecA overexpression is neither toxic nor is the expression of the recA(+) gene affected in a recX deletion strain at the basal level or after UV induction. PMID- 12547391 TI - Overexpression of Cu/Zn superoxide dismutase is lethal for mice lacking double strand break repair. AB - The non-homologous DNA end joining (NHEJ) pathway is a major double-strand DNA break repair pathway in cells of multicellular eukaryotes. Ku is a heterodimeric protein consisting of Ku70 and Ku86, and it is thought to be the first component to bind to a broken double-strand DNA end. Mice lacking Ku86 show features of premature aging, live about 6-12 months, and show a characteristic loss of neurons in the central nervous system during development. Cells from mice lacking Ku have increased numbers of chromosome breaks, a significant fraction of which are caused by oxidative metabolism. Overexpression of the cytoplasmic Cu/Zn superoxide dismutase (SOD1) from a transgene is known to increase the number of chromosome breaks in primary cells (presumably by increasing reactive oxygen species). Here we show that SOD1 overexpression in a Ku86-/- mouse results in embryonic lethality. This striking effect is, however, subject to a strain specific modifier. Genome-wide marker analysis is most consistent with the modifier being on mouse chromosome 13. Analysis of 10 markers on chromosome 13 suggests that the modifier is within the same region as a modifier of the murine amyotropic lateral sclerosis (ALS) phenotype when it is caused by overexpression of a mutant form of SOD1. Based on these results, we propose a model in which oxidative metabolism causes chromosome breaks, leading to neuronal death; and this neuronal death may account for that seen in NHEJ mutant animals and in mammals with SOD1-mediated ALS. PMID- 12547392 TI - Caloric restriction promotes genomic stability by induction of base excision repair and reversal of its age-related decline. AB - Caloric restriction is a potent experimental manipulation that extends mean and maximum life span and delays the onset and progression of tumors in laboratory rodents. While caloric restriction (CR) clearly protects the genome from deleterious damage, the mechanism by which genomic stability is achieved remains unclear. We provide evidence that CR promotes genomic stability by increasing DNA repair capacity, specifically base excision repair (BER). CR completely reverses the age-related decline in BER capacity (P<0.01) in all tissues tested (brain, liver, spleen and testes) providing aged, CR animals with the BER phenotype of young, ad libitum-fed animals. This CR-induced reversal of the aged BER phenotype is accompanied by a reversal in the age-related decline in DNA polymerase beta (beta-pol), a rate-limiting enzyme in the BER pathway. CR significantly reversed the age-related loss of beta-pol protein levels (P<0.01), mRNA levels (P<0.01) and enzyme activity (P<0.01) in all tissues tested. Additionally, in young (4-6 month-old) CR animals a significant up-regulation in BER capacity, beta-pol protein and beta-pol mRNA is observed (P<0.01), demonstrating an early effect of CR that may provide insight in distinguishing the anti-tumor from the anti-aging effects of CR. This up-regulation in BER by caloric restriction in young animals corresponds to increased protection from carcinogen exposure, as mutation frequency is significantly reduced in CR animals exposed to either DMS or 2 nitropropane (2-NP) (P<0.01). Overall the data suggest an important biological consequence of moderate BER up-regulation and provides support for the hormesis theory of caloric restriction. PMID- 12547393 TI - Induction of DNA double strand breaks by arsenite: comparative studies with DNA breaks induced by X-rays. AB - Chinese hamster ovary (CHO-K1) cell line and two of its DNA double strand break (DSB) repair deficient mutant cell lines, xrs-5 (Ku80 mutant) and irs-20 (DNA PKcs mutant), were treated with various concentrations of sodium arsenite for 2.5h, and the colony forming abilities were studied. The wild type cells showed the highest cell survival, while xrs-5 cells showed the lowest survival, and irs 20 cells had an intermediate survival. These results are very similar to the cell survival curves induced by X-rays in these three cell lines. Our data also show the dose dependent induction of DNA-DSBs in these cell lines exposed to arsenite. However, in order to obtain a similar cell survival in wild type cells, twice as many DNA-DSBs are necessary with arsenite exposure when compared with X-rays, suggesting that the types of DNA lesions leading to DSB induced by arsenite are different from those by X-rays. Based on these data, further mechanistic investigations including the involvement of DNA-DSB repair proteins are warranted in the recovery process from arsenic (As) exposure. PMID- 12547394 TI - hSMUG1 can functionally compensate for Ung1 in the yeast Saccharomyces cerevisiae. AB - There are at least four distinct families of enzymes that recognize and remove uracil from DNA. Family-3 (SMUG1) enzymes have recently been identified and have a preference for uracil in single-stranded DNA when assayed in vitro. Here we investigate the in vivo function of SMUG1 using the yeast Saccharomyces cerevisiae as a model system. These organisms lack a SMUG1 homologue and use a single enzyme, Ung1 to carry out uracil-repair. When a wild-type strain is treated with antifolate agents to induce uracil misincorporation into DNA, S phase arrest and cellular toxicity occurs. The arrest is characteristic of checkpoint activation due to single-strand breaks caused by continuous uracil removal and self-defeating DNA repair. When uracil-DNA glycosylase is deleted (deltaung1), cells continue through S-phase and arrest at G(2)/M, presumably due to the effects of stable uracil misincorporation in DNA. Pulsed field gel electrophoresis (PFGE) demonstrates that cells are able to complete DNA replication with uracil-substituted DNA and do not experience the extensive strand breakage attributed to uracil-DNA glycosylase-mediated repair. As a result, these cells experience early protection from antifolate-induced cytotoxicity. When either UNG1 or SMUG1 functions are reintroduced back into the null strain and then subjected to antifolate treatment, the cells revert back to the wild-type phenotype as shown by a restored sensitivity to drug and S-phase arrest. The arrest is accompanied by the accumulation of replication intermediates as determined by PFGE. Collectively, these data indicate that SMUG1 can act as a functional homolog of the family-1 uracil-DNA glycosylase enzymes. PMID- 12547395 TI - DNA bending by the human damage recognition complex XPC-HR23B. AB - Genome integrity is maintained, despite constant assault on DNA, due to the action of a variety of DNA repair pathways. Nucleotide excision repair (NER) protects the genome from the deleterious effects of UV irradiation as well as other agents that induce chemical changes in DNA bases. The mechanistic steps required for eukaryotic NER involve the concerted action of at least six proteins or protein complexes. The specificity to incise only the DNA strand including the damage at defined positions is determined by the coordinated assembly of active protein complexes onto damaged DNA. In order to understand the molecular mechanism of the NER reactions and the origin of this specificity and control we analyzed the architecture of functional NER complexes at nanometer resolution by scanning force microscopy (SFM). In the initial step of damage recognition by XPC HR23B we observe a protein induced change in DNA conformation. XPC-HR23B induces a bend in DNA upon binding and this is stabilized at the site of damage. We discuss the importance of the XPC-HR23B-induced distortion as an architectural feature that can be exploited for subsequent assembly of an active NER complex. PMID- 12547396 TI - Isolation and genetic characterization of the Neurospora crassa REV1 and REV7 homologs: evidence for involvement in damage-induced mutagenesis. AB - In a previous paper, we reported that the Neurospora crassa upr-1 gene is a homolog of the yeast gene REV3, which encodes the catalytic subunit of DNA polymerase zeta (polzeta). Characterization of the upr-1 mutant indicated that the UPR1 protein plays a role in DNA repair and mutagenesis. To help understand the mechanisms of mutagenic DNA repair in the N. crassa more extensively, we identified N. crassa homologs of yeast REV1 and REV7 and obtained mutants ncrev1 or ncrev7, which had similar phenotypes to the upr-1 mutant. Mutant carrying ncrev7 was more sensitive to UV and 4NQO, and slightly sensitive to MMS than the wild-type. The sensitivity to UV and MMS of the ncrev1 mutant was moderately higher than that of the wild-type, but the sensitivity to 4NQO of the mutant was similar to that of the wild-type. In reversion assay using testers with base substitution or frameshift mutation at the ad-3A locus, each of ncrev1 and ncrev7 mutants showed lower induced-mutability than the wild-type. Expression of ncrev1 and ncrev7 was found to be UV-inducible like the case of upr-1. Genetic analyses showed that the ncrev7 was identical to mus-26, which belongs to the upr-1 epistasis group, and that the ncrev1 was a newly identified DNA repair gene and designated as mus-42. Interestingly, all three mutants have a normal CPD photolyase gene, however, they showed a partial photoreactivation defect (PPD) phenotype, not completely defective but inefficient in photoreactivation. These results suggest that N. crassa REV homolog genes function in DNA repair and UV mutagenesis through the bypass of (6-4) photoproducts. PMID- 12547397 TI - Budding yeast mms4 is epistatic with rad52 and the function of Mms4 can be replaced by a bacterial Holliday junction resolvase. AB - MMS4 of Saccharomyces cerevisiae was originally identified as the gene responsible for one of the collection of methyl methanesulfonate (MMS)-sensitive mutants, mms4. Recently it was identified as a synthetic lethal gene with an SGS1 mutation. Epistatic analyses revealed that MMS4 is involved in a pathway leading to homologous recombination requiring Rad52 or in the recombination itself, in which SGS1 is also involved. MMS sensitivity of mms4 but not sgs1, was suppressed by introducing a bacterial Holliday junction (HJ) resolvase, RusA. The frequencies of spontaneously occurring unequal sister chromatid recombination (SCR) and loss of marker in the rDNA in haploid mms4 cells and interchromosomal recombination between heteroalleles in diploid mms4 cells were essentially the same as those of wild-type cells. Although UV- and MMS-induced interchromosomal recombination was defective in sgs1 diploid cells, hyper-induction of interchromosomal recombination was observed in diploid mms4 cells, indicating that the function of Mms4 is dispensable for this type of recombination. PMID- 12547400 TI - Autoimmune hepatitis: "Hepatitis sui generis". PMID- 12547402 TI - Down-regulation of the Na+/taurocholate cotransporting polypeptide during pregnancy in the rat. AB - BACKGROUND: Experimental studies have shown decreased bile acid (BA) uptake and reduced excretion of cholephilic compounds in pregnant rodents. AIM: To assess the expression and function of the main BA importer, the Na(+)/taurocholate cotransporting polypeptide (Ntcp) in pregnant rats. METHODS: BA uptake and Ntcp expression were studied in control and timed-pregnant rats in late gestation. Ntcp protein, messenger RNA (mRNA) expression, and Ntcp tissue localization were determined by Northern blotting, Western analysis, and tissue immunofluorescence. The activity of three transactivators of the Ntcp promoter: hepatocyte nuclear factor 1-alpha (HNF1-alpha), nuclear receptor heterodimer retinoid X receptor:retinoid acid receptor (RXR:RAR) and signal transducer and activator of transcription 5 (Stat5) was assessed using gel electrophoretic mobility shift assays. RESULTS: A significantly reduced BA uptake and decreased Ntcp mRNA levels (-40%) and protein mass (-60%) was observed in pregnant rats. Nuclear extracts from pregnant rats showed a marked decrease of HNF1-alpha and RXR:RAR binding activities by -80 and -40% of basal activity, respectively. In contrast, binding activity of Stat-5 was increased by 50% in nuclear extracts from pregnant rats. CONCLUSIONS: Pregnancy is associated with reduced Ntcp expression and function in the rat. Our findings suggest that Ntcp down-regulation during pregnancy occurs primarily at the transcriptional level. PMID- 12547403 TI - The hepatocyte is a direct target for transforming-growth factor beta activation via the insulin-like growth factor II/mannose 6-phosphate receptor. AB - BACKGROUND/AIMS: The cation-independent mannose 6-phosphate receptor (CIMPR) is overexpressed in hepatocytes during liver regeneration and has been implicated in the maturation of latent pro-transforming growth factor beta (TGFbeta). In this study, we have: (1) kinetically characterized the changes in CIMPR expression in regenerating liver and cultured proliferating hepatocytes; and (2) assessed the contribution of hepatocyte via the CIMPR to latent pro-TGFbeta activation. METHODS: The expression of CIMPR protein and mRNA in livers collected after partial hepatectomy and hepatocyte primary cultures was analyzed by Western and Northern blotting. Activity of latent pro-TGFbeta was assessed by inhibition of [3H] methylthymidine incorporation into DNA. RESULTS: The expression of the CIMPR protein and/or mRNA progressively increased after 8 h in regenerating liver and 42-46 h in cultured hepatocytes, prior to the onset of DNA replication. Both mature TGFbeta and latent pro-TGFbeta inhibited epidermal growth factor stimulated DNA synthesis in hepatocytes in a dose-dependent manner. The effect of latent pro-TGFbeta was reversed by two ligands of the CIMPR: beta-galactosidase, a mannose 6-phosphate containing protein, and a CIMPR antibody. CONCLUSIONS: (1) The induction of the CIMPR gene during liver regeneration and hepatocyte culture occurs in mid G1 phase; and (2) the CIMPR mediates latent proTGFbeta activation and thus may act, by targeting TGFbeta to hepatocytes, as a negative regulator of hepatocyte growth. PMID- 12547401 TI - Taurohyodeoxycholate- and tauroursodeoxycholate-induced hypercholeresis is augmented in bile duct ligated rats. AB - BACKGROUND/AIMS: Taurohyodeoxycholate (THDCA) and tauroursodeoxycholate (TUDCA) induce more bile flow per molecule excreted compared to endogenous bile acids. The aim of this study is to determine if the hypercholeretic effect of tauroursodeoxycholate or taurohyodeoxycholate in normal and bile duct ligated (BDL) rats is due to increased ductal secretion. METHODS: Normal or BDL rats were infused with tauroursodeoxycholate or taurohyodeoxycholate and bile flow, bicarbonate, bile salt, cholesterol, and phospholipid secretion were measured. Cholangiocytes were stimulated with taurohyodeoxycholate or tauroursodeoxycholate, and secretin-stimulated secretion was measured. RESULTS: Taurohyodeoxycholate and tauroursodeoxycholate increased bile flow more in BDL than normal rats. Tauroursodeoxycholate increased bicarbonate secretion more in BDL compared to normal rats. Taurohyodeoxycholate when infused with taurocholate increased bile flow (but not phospholipid excretion) to a greater degree in BDL compared to normal rats. Taurohyodeoxycholate and tauroursodeoxycholate decreased secretin-stimulated cholangiocyte secretion. CONCLUSIONS: Consistent with a ductal origin for bile acid-induced hypercholeresis, taurohyodeoxycholate and tauroursodeoxycholate produced a greater hypercholeresis in BDL than normal rats. Tauroursodeoxycholate- (but not taurohyodeoxycholate-) stimulated hypercholeresis is associated with increased HCO(3)(-) secretion. Tauroursodeoxycholate increases biliary HCO(3)(-) secretion by a mechanism unrelated to secretin-stimulated cholangiocyte secretion. Taurohyodeoxycholate-induced hypercholeresis in BDL rats is unrelated to enhanced phospholipid excretion. PMID- 12547404 TI - The canine copper toxicosis gene MURR1 does not cause non-Wilsonian hepatic copper toxicosis. AB - BACKGROUND: Non-Wilsonian hepatic copper toxicosis includes Indian childhood cirrhosis (ICC), endemic Tyrolean infantile cirrhosis (ETIC) and the non-Indian disease known as idiopathic copper toxicosis (ICT). These entities resemble the hepatic copper overload observed in livers of Bedlington terriers with respect to their clinical presentation and biochemical and histological findings. We recently cloned the gene causing copper toxicosis in Bedlington terriers, MURR1, as well as the orthologous human gene on chromosome 2p13-p16. AIM: To study the human orthologue of the canine copper toxicosis gene as a candidate gene for ICC, ETIC, and ICT. METHODS: We sequenced the exons and the intron-exon boundaries of the human MURR1 gene in 12 patients with classical ICC, one patient with ETIC, and 10 patients with ICT to see whether these patients display any mutations in the human orthologue of the canine copper toxicosis gene. RESULTS: No mutations in the MURR1 gene, including the intron-exon boundaries, were identified in a total of 23 patients with non-Wilsonian hepatic copper toxicosis. CONCLUSIONS: Our results demonstrate that copper toxicosis in Bedlington terriers is not an animal model for the non-Wilsonian hepatic copper toxicosis described in this study. PMID- 12547405 TI - Adenoviral-mediated transfer of p53 or retinoblastoma protein blocks cell proliferation and induces apoptosis in culture-activated hepatic stellate cells. AB - BACKGROUND/AIMS: The principal cellular effectors of fibrosis in liver are hepatic stellate cells (HSC). In response to liver injury these quiescent cells undergo a phenotypic change to a myofibroblastic cell type, proliferate and secrete matrix components. Thus, removal of activated HSC should be beneficial for the prognosis of hepatic fibrogenesis and preserve organ function. METHODS: The purpose of this study was to investigate whether administration of adenoviruses constitutively expressing the p53 tumor suppressor or the retinoblastoma protein (pRb) is sufficient to induce cell arrest or apoptosis in culture-activated HSC. The expression of the transgenes was confirmed by Western blot analysis and immunohistochemistry. RESULTS: Both proteins were expressed mainly in the nucleus and their expression was associated with a marked inhibition of cell proliferation and induction of apoptosis as determined by measurement of phosphatidylserine exposed at the surface, proliferation assay, induction of the p21 cyclin-dependent kinase inhibitor, and an increase of caspase-3 activity. Additionally, electron microscopic analysis confirmed that activation of the p53-mediated pathway in HSC results in chromatin and cytoplasmic condensation, typical features of ongoing apoptosis. CONCLUSIONS: Our results indicate that transduction of p53 or pRb offers a feasible approach to induce apoptosis in activated HSC. Thus, targeted transfer of these proteins into HSC may be potentially useful for the treatment of hepatic fibrosis. PMID- 12547406 TI - Selective blockade of mGlu5 metabotropic glutamate receptors is protective against acetaminophen hepatotoxicity in mice. AB - BACKGROUND/AIMS: mGlu5 metabotropic glutamate receptor antagonists protect rat hepatocytes against hypoxic death. Here, we have examined whether mGlu5 receptor antagonists are protective against liver damage induced by oxidative stress. METHODS: Toxicity of isolated hepatocytes was induced by tert-butylhydroperoxide (t-BuOOH) after pretreatment with the mGlu5 receptor antagonists, MPEP, SIB-1757 and SIB-1893. The effect of these drugs was also examined in mice challenged with toxic doses of acetaminophen. RESULTS: Addition of tBuOOH (0.5 mM) to isolated hepatocytes induced cell death (70+/-5% at 3 h). Addition of MPEP or SIB-1893 to hepatocytes reduced both the production of reactive oxygen species (ROS) and cell toxicity induced by t-BuOOH (tBuOOH=70+/-5%; tBuOOH+MPEP=57+/-6%; tBuOOH+SIB 1893=40+/-4%). In mice, a single injection of acetaminophen (300 mg/kg, i.p.) induced centrilobular liver necrosis, which was detectable after 24 h. MPEP (20 mg/kg, i.p.) substantially reduced liver necrosis and the production of ROS, although it did not affect the conversion of acetaminophen into the toxic metabolite, N-acetylbenzoquinoneimine. MPEP, SIB-1893 and SIB-1757 (all at 20 mg/kg, i.p.) also reduced the increased expression and activity of liver iNOS induced by acetaminophen. CONCLUSIONS: We conclude that pharmacological blockade of mGlu5 receptors might represent a novel target for the treatment of drug induced liver damage. PMID- 12547407 TI - Cerebral oxygenation determined by near-infrared spectrophotometry in patients with fulminant hepatic failure. AB - BACKGROUND/AIMS: In severe cases of acute liver failure (ALF), cerebral hyperperfusion may result in high intracranial pressure and brain damage. The aim of this study was to determine if near-infrared spectrophotometry (NIRS) could detect a raise in cerebral blood flow and oxygenation induced by noradrenaline (NA) infusion. METHODS: In seven ALF patients (five females and two males; median age 49 years (range 20-70)) changes in cerebral concentration of oxy (deltaHbO(2)) and total-haemoglobin (deltaHbT) were compared to the jugular bulb saturation (SvjO(2)) and cerebral blood flow velocity (Vmean) during NA infusion. RESULTS: Mean arterial pressure increased from 68 (64-86) to 103 (87-118) mmHg and the cerebral perfusion pressure from 61 (53-79) to 95 (74-110) mmHg (P<0.05), while the intracranial pressure (7 (6-15) mmHg) was not significantly changed. In six patients cerebral deltaHbO(2) and deltaHbT increased 2.7 (0.3-9.6) and 2.0 (0.3-14.8) micromol l(-1), respectively, but cerebral oxygenation decreased in one patient. SvjO(2) increased from 68 (55-76) to 74 (64-78) % (P<0.05) concomitant with an increase in Vmean from 47 (34-65) to 68 (50-86) cm s(-1) (P<0.05). deltaHbO(2) covariated with changes in SvjO(2) during NA in all but one patient. CONCLUSIONS: In ALF patients, a change in cerebral perfusion was detected by NIRS. The combination of NIRS and transcranial Doppler sonography may be valuable non-invasive techniques to detect cerebral hyperperfusion before intracranial hypertension becomes manifest. PMID- 12547408 TI - Kupffer cells modulate splenic interleukin-10 production in endotoxin-induced liver injury after partial hepatectomy. AB - BACKGROUND/AIMS: This study was conducted to investigate the implication of Kupffer cells and the spleen in interleukin (IL)-10 production in endotoxin induced liver injury after hepatectomy. METHODS: Rats were divided into five groups: the S group, sham-operation; the SG group, sham-operation followed by intravenous gadolinium chloride (GdCl(3): 7 mg/kg) administration to inhibit Kupffer cell function; the H group, two-thirds hepatectomy; the HG group, hepatectomy and subsequent GdCl(3) administration; the HGS group, hepatectomy and splenectomy with GdCl(3) administration. Lipopolysaccharide (1.5 mg/kg) was intravenously administered for each group 48 h after surgery. RESULTS: GdCl(3) treatment significantly suppressed the elevation of plasma tumor necrosis factor (TNF)-alpha levels by lipopolysaccharide administration with completely inhibited induction of hepatic TNF-alpha and IL-10 mRNAs. In the HG group, marked increase in plasma IL-10 levels associated with enhanced splenic IL-10 mRNA was observed 1 h after lipopolysaccharide administration when compared to those in the H and HGS groups. Plasma TNF-alpha/IL-10 ratio 1 h after lipopolysaccharide administration was higher in the order of H, HGS and HG groups. Hepatic parenchymal damage and the 24-h mortality were lowest in group HG, followed by groups HGS and H. CONCLUSIONS: Kupffer cells after hepatectomy may aggravate endotoxin-induced liver injury via down-regulation of IL-10 production in the spleen. PMID- 12547409 TI - Risk factors contributing to early and late phase intrahepatic recurrence of hepatocellular carcinoma after hepatectomy. AB - BACKGROUND/AIMS: We conducted a retrospective cohort study to investigate factors to early and late phase recurrence of hepatocellular carcinoma (HCC). METHODS: The study population consisted of 249 patients including 157 with cirrhosis who underwent hepatectomy for HCC. The endpoint was time-to-recurrence. Using a Cox regression model, factors to early and late phase recurrences were investigated censoring recurrence-free patients at the 2-year time point and in patients without recurrence at 2 years. RESULTS: Actuarial probability of overall recurrence at 1, 3, and 5 years were 0.301, 0.623, and 0.790, respectively, with a median follow-up of 624 days. Early recurrence was observed in 123 out of 249 patients; while late recurrence was found in 61 out of 113 patients. Factors to early recurrence were as follows: non-anatomical resection, presence of microscopic vascular invasion, and serum alpha-fetoprotein level >or=32 ng/ml. Those contributing to late phase recurrence were higher grade of hepatitis activity, multiple tumors, and gross tumor classification. CONCLUSIONS: Variables associated with metastatic recurrence were factors to early phase recurrence; whereas those related with elevated carcinogenesis contributed to late phase recurrence, thus providing an epidemiological evidence that different mechanisms, i.e. metastasis and de novo, are involved in intrahepatic recurrence after hepatectomy for HCC. PMID- 12547410 TI - Hepatic immunohistochemical staining with a monoclonal antibody against HCV-E2 to evaluate antiviral therapy and reinfection of liver grafts in hepatitis C viral infection. AB - BACKGROUND/AIMS: A simple and reproducible hepatic immunohistochemical staining (IHS) for hepatitis C virus (HCV) is not available. We aimed to validate hepatic IHS with monoclonal antibody (Mab) IG222, directed against the HCV-envelope 2 (E2) protein. METHODS: A three-step indirect immunoperoxidase method was used for frozen sections and a two-step indirect EnVision technique was used for paraffin embedded sections. RESULTS: Naturally or in vitro HCV infected primary human hepatocytes were immunoreactive to HCV-E2. In the patient study (n=253), IHS had a sensitivity of 96% and a specificity of 91%. Six patients who showed positivity in the liver with Mab IG222, but remained anti-HCV and HCV-RNA negative, had hepatitis C-like changes in their liver biopsy. In one patient HCV-RNA could be detected in the liver biopsy. We confirmed early graft reinfection in patients transplanted for HCV-related disease (34 patients with serial biopsies). Treatment for acute cellular rejection with steroids was associated with an increase in staining intensity. In nine patients with clearance of HCV-RNA during antiviral therapy, seven achieved negativation of immunoreactivity and two a marked reduction. CONCLUSIONS: The IHS with Mab IG222 is an accurate tool for diagnosis and clinical management of chronic hepatitis C. PMID- 12547411 TI - Assessment of cost-effectiveness of universal hepatitis B immunization in a low income country with intermediate endemicity using a Markov model. AB - BACKGROUND/AIMS: Most countries with high hepatitis B (HB) virus endemicity and most high-income countries have introduced immunization programmes against this infection. However, several low-income countries with intermediate HB endemicity have not done so. We performed a cost-effectiveness analysis of universal childhood HB immunization in such countries using India as an example, since available data on this aspect are limited. METHODS: Marginal cost of every life year and quality-adjusted life-year (QALY) gained with universal HB vaccination was calculated using a Markov model. Two types of analyses (including and excluding expenditure on treatment of long-term complications of HB infection) were done. Several sensitivity analyses and Monte-Carlo simulation were performed. RESULTS: Universal immunization reduced the HB carrier rate by 71%, and increased the number of years and QALY lived by a birth-cohort by 0.173 years (61.072 vs. 60.899 years) and 0.213 years (61.056 vs. 60.843 years), respectively. Marginal costs were US$16.27 per life-year gained and US$13.22 per QALY gained, much lower than annual per capita income. One-way sensitivity analysis and Monte-Carlo simulation confirmed the robustness of the conclusions. CONCLUSIONS: Universal HB immunization is highly cost-effective in low-income countries with intermediate endemicity rates. PMID- 12547412 TI - Histopathological study of chronic hepatitis B and C: a comparison of two scoring systems. AB - BACKGROUND/AIMS: Several histological scoring systems are used to evaluate chronic viral hepatitis. This study was undertaken to determine the correlation between the Ishak system (modified histological activity index, HAI) and the METAVIR system, in Indian patients with chronic viral hepatitis. METHODS: Liver biopsies from 127 patients with chronic viral hepatitis B or C were examined, and scored using the Ishak and METAVIR systems, and weighted kappa analysis of correlation was done. Correlation of necroinflammatory activity with serum transaminase levels was analyzed, and prevalence of specific histological features compared in hepatitis B virus (HBV) and HCV biopsies. RESULTS: HBV infection accounted for 64.6% of cases, and HCV for 35.4%; 91.3% of patients had minimal or mild hepatitis. The necroinflammatory scores of the Ishak and METAVIR systems correlated moderately well (weighted kappa 0.627), while there was excellent correlation with regard to fibrosis (weighted kappa 0.998). Similar concordance was found when HBV and HCV cases were analyzed separately. HAI showed poor correlation with serum transaminases (weighted kappa 0.21). Micronodular cirrhosis, lymphoid aggregates, bile duct damage, bile ductular proliferation and steatosis were significantly more common in HCV biopsies compared to HBV. CONCLUSIONS: Concordance between Ishak and METAVIR scoring systems is good for necroinflammatory change, and excellent for fibrotic change. PMID- 12547413 TI - Enhanced monocyte Th1 cytokine production in HCV-infected cryoglobulinemic patients. AB - BACKGROUND/AIMS: The etiologic link between chronic hepatitis C virus (HCV) infection and mixed cryoglobulinemia is well established, while its prognostic significance within the context of HCV-related hepatitis is not as clear. Patients with an HCV-related cryoglobulinemic syndrome oft have mild liver disease, an aspect that can be influenced by an individual's Th1/Th2 orientation. Our goal was to document stigmata of differentiate cytokine production in this subgroup of patients. METHODS: Fifteen patients with chronic HCV-related liver disease (CLD) and a cryoglobulinemic syndrome (CRYO) were compared to age/sex matched CLD controls with negative cryocrit. Cultured monocytes were stimulated with either Staphylococcus aureus (SAC) or lipopolysaccharide (LPS). RESULTS: The protein concentrations of TNF-alpha and of the Th1-type cytokines interleukin (IL)-12 and IL-18 were significantly greater in the CRYO group, while IL-10 (a Th2 cytokine) levels were greater in the control group. CONCLUSIONS: The clinical distinctiveness of the two groups was reflected at the cytokine level. The cryoglobulinemic patients studied showed a greater Th1 polarization than their cryoglobulin-negative counterparts. This enhanced production of Th1-type cytokines is seemingly not able to rid the host of infection but may account for a milder course of liver disease. PMID- 12547414 TI - Prognostic factors after resection of hepatocellular carcinoma: are there landmarks in the wild forest? PMID- 12547415 TI - Assessment of liver biopsies in chronic hepatitis: how is it best done? PMID- 12547416 TI - Hepatic osteodystrophy. PMID- 12547417 TI - Lack of hepatitis C transmission among institutionalized psychiatric patients. PMID- 12547418 TI - In vino veritas? PMID- 12547420 TI - Domains, motifs and clusters in the protein universe. AB - The rapid growth of bio-sequence information has resulted in an increasing demand for reliable methods that group proteins. A few databases with curated alignments of protein families have demonstrated that expert-driven repositories can keep up with the data deluge in the genome era. These original resources implicitly identify domain-like modules in proteins. An increasing number of automatic methods have sprouted over the past few years that cluster the protein universe. Many of these implicitly dissect proteins into structural domain-like fragments. In a very coarse-grained evaluation, some of the automatic methods appear to be on par with expert-driven approaches. However, neither automatic nor manual methods are currently entirely up to the challenges of tasks such as target selection in structural genomics. Thus, we urgently need refined and sustained automatic clustering tools. PMID- 12547421 TI - Emergence of diverse biochemical activities in evolutionarily conserved structural scaffolds of proteins. AB - Comparative analysis of numerous protein structures that have become available in the past few years, combined with genome comparison, has yielded new insights into the evolution of enzymes and their functions. In addition to the well-known diversification of substrate specificities, enzymes with several widespread catalytic folds, particularly the TIM barrel, the RRM-like domain and the double stranded beta-helix (cupin) domain, have been extensively explored in 'reaction space', resulting in the evolution of numerous, diverse catalytic activities supported by the same structural scaffold. Common protein folds differ widely in the diversity of catalyzed reactions. The biochemical plasticity of a fold seems to hinge on the presence of a generic, symmetrical substrate-binding pocket as opposed to highly specialized binding sites. PMID- 12547422 TI - Protein complexes and proteome organization from yeast to man. AB - Protein complexes may well be the most relevant molecular units of cellular function. The activities of protein complexes have to be regulated both in time and space to integrate within the overall cell programs. The cell can be compared to a factory orchestrating individual assembly lines into integrated networks fulfilling particular and superimposed tasks. Recent proteome-wide studies provide insight into the properties of cellular protein complexes, their modular nature, their interaction with other complexes and the resulting preliminary organization chart of the proteome. PMID- 12547423 TI - Overview of structural genomics: from structure to function. AB - The unprecedented increase in the number of new protein sequences arising from genomics and proteomics highlights directly the need for methods to rapidly and reliably determine the molecular and cellular functions of these proteins. One such approach, structural genomics, aims to delineate the total repertoire of protein folds, thereby providing three-dimensional portraits for all proteins in a living organism and to infer molecular functions of the proteins. The goal of obtaining protein structures on a genomic scale has motivated the development of high-throughput technologies for macromolecular structure determination, which have begun to produce structures at a greater rate than previously possible. These new structures have revealed many unexpected functional and evolution relationships that were hidden at the sequence level. PMID- 12547424 TI - Genetic screens and directed evolution for protein solubility. AB - Overexpressed proteins are often insoluble, and can be recalcitrant to conventional solubilization techniques such as refolding. Directed evolution methods, in which protein diversity libraries are screened for soluble variants, offer an alternative route to obtaining soluble proteins. Recently, several new protein solubility screens have been developed that do not require structural or functional information about the target protein. Soluble protein can be detected in vivo and in vitro by fusion reporter tags. Protein misfolding can be measured in vivo using the bacterial response to protein misfolding. Finally, soluble protein can be monitored by immunological detection. Efficient, well-established strategies for generating and recombining genetic diversity, driven by new screening and selection methods, can furnish correctly folded, soluble protein. PMID- 12547425 TI - Protein expression systems for structural genomics and proteomics. AB - One of the key steps of structural genomics and proteomics is high-throughput expression of many target proteins. Gene cloning, especially by ligation independent cloning techniques, and recombinant protein expression using microbial hosts such as Escherichia coli and the yeast Pichia pastoris are well optimized and further robotized. Cell-free protein synthesis systems have been developed for large-scale production of protein samples for NMR (stable-isotope labeling) and X-ray crystallography (selenomethionine substitution). Protein folding is still a major bottleneck in protein expression. Cell-based and cell free methods for screening of suitable samples for structure determination have been developed for achieving a high success rate. PMID- 12547426 TI - Ontologies for proteomics: towards a systematic definition of structure and function that scales to the genome level. AB - A principal aim of post-genomic biology is elucidating the structures, functions and biochemical properties of all gene products in a genome. However, to adequately comprehend such a large amount of information we need new descriptions of proteins that scale to the genomic level. In short, we need a unified ontology for proteomics. Much progress has been made towards this end, including a variety of approaches to systematic structural and functional classification and initial work towards developing standardized, unified descriptions for protein properties. In relation to function, there is a particularly great diversity of approaches, involving placing a protein in structured hierarchies or more generalized networks and a recent approach based on circumscribing a protein's function through systematic enumeration of molecular interactions. PMID- 12547427 TI - Protein chip technology. AB - Microarray technology has become a crucial tool for large-scale and high throughput biology. It allows fast, easy and parallel detection of thousands of addressable elements in a single experiment. In the past few years, protein microarray technology has shown its great potential in basic research, diagnostics and drug discovery. It has been applied to analyse antibody-antigen, protein-protein, protein-nucleic-acid, protein-lipid and protein-small-molecule interactions, as well as enzyme-substrate interactions. Recent progress in the field of protein chips includes surface chemistry, capture molecule attachment, protein labeling and detection methods, high-throughput protein/antibody production, and applications to analyse entire proteomes. PMID- 12547428 TI - Tackling the phosphoproteome: tools and strategies. AB - Characterization of post-translational modifications in proteins is one of the major tasks that is to be accomplished in the post-genomic era. Phosphorylation is a key reversible modification that regulates enzymatic activity, subcellular localization, complex formation and degradation of proteins. Rapid advances in mass spectrometry instrumentation coupled to the development of analytical methods over the past several years now allow us to investigate the phosphoproteome on a global scale. PMID- 12547429 TI - Quantitative proteomics using mass spectrometry. AB - The use of stable isotopes as internal standards in mass spectrometry has opened a new era for quantitative proteomics. Depending on the point at which the label is introduced, most procedures can be classified as in vivo labeling, in vitro pre-digestion labeling or in vitro post-digestion labeling. In vivo labeling has been used for cells that can be grown in culture and has the advantage of being more accurate. The pre-digestion and post-digestion labeling procedures are suitable for all types of sample including human body fluids and biopsies. Several new mass spectrometric strategies mark significant achievements in determining relative protein concentrations and in quantifying post-translational modifications. However, further technology developments are needed for understanding the complexity of a dynamic system like the proteome. PMID- 12547430 TI - Activity-based proteomics: enzyme chemistry redux. AB - The principles of enzyme chemistry, mechanism of action and inhibitor design are being applied to proteomics by the development of activity-based probes. This approach suggests a potentially broad method for interrogating enzyme family members, both known and unknown, in cells and proteomic fractions without the need for individual assay development and isolation. The serine hydrolases and cysteine proteases have provided the proofs of concept for activity-based proteomics, and other studies are rapidly following. The result will be a proteomics technology of great value to drug discovery and development. PMID- 12547431 TI - Peptide libraries: at the crossroads of proteomics and bioinformatics. AB - Peptide libraries offer a valuable means for providing functional information regarding protein-modifying enzymes and protein interaction domains. Library approaches have become increasingly useful as high-throughput strategies for the analysis of large numbers of new proteins identified as a result of genome sequencing efforts. Recent developments in the field have produced faster methods with broadened applicability. Crucially, new computational and biochemical tools have emerged that facilitate identification of interaction partners and substrates for proteins on the basis of their peptide selectivity profiles. Such combinations of proteomics-scale experimental approaches with bioinformatics tools hold great promise for the elucidation of protein interaction networks and signal transduction pathways in living cells. PMID- 12547432 TI - Forward chemical genetics: progress and obstacles on the path to a new pharmacopoeia. AB - Forward chemical genetics is a new method to systematize the discovery and use of small molecules as tools for basic biological research. This approach requires three basic components: a library of compounds; an assay, in which the library is screened for a cellular or organismal phenotype; and a method to trace an active compound to its biological target. Bioactive compounds have traditionally been isolated from natural product extracts, although 'diversity-oriented synthesis' and commercial compound collections are gaining in prominence. New techniques, such as image-based screening and the cytoblot method, have increased the throughput of phenotypic assays. Strategies are also being developed to streamline target identification using molecular biological approaches. PMID- 12547433 TI - Functional genomics of intracellular peptide recognition domains with combinatorial biology methods. AB - Phage-displayed peptide libraries have been used to identify specific ligands for peptide-binding domains that mediate intracellular protein-protein interactions. These studies have provided significant insights into the specificities of particular domains. For PDZ domains that recognize C-terminal sequences, the information has proven useful in identifying natural binding partners from genomic databases. For SH3 domains that recognize internal proline-rich motifs, the results of database searches with phage-derived ligands have been compared with the results of yeast-two-hybrid experiments to produce overlap networks that reliably predict natural protein-protein interactions. In addition, libraries of phage-displayed PDZ and SH3 domains have been used to identify the residues responsible for ligand recognition, and also to engineer domains with altered specificities. PMID- 12547435 TI - Computational bioinorganic chemistry. Part III. The tools of the trade: from high level ab initio calculations to structural bioinformatics. AB - Having long focused on the electronic-structural aspects of metalloenzymes and their synthetic models, Abhik is currently exploring a number of synthetic problems related to self-assembly processes, dynamic combinatorial libraries, and fluorine chemistry. PMID- 12547434 TI - Genome-wide analysis of signaling domain function. AB - Approximately 2.5% of human gene products contain one or more small domains that drive interactions between proteins and other cellular components in cell signaling processes. The many interactions driven by these relatively simple domains are thought to cooperate with one another to yield complex signaling networks that allow very fine control of cell function. In principle, if we can understand all domain-mediated interactions it should be possible to model these networks. Genome-wide analysis of signaling domain interactions represents a first step in this direction, and several advances of this sort in yeast have been reported over the past year. These reports suggest, for some domains at least, that the prospect of generating 'wiring diagrams' with this simple approach is feasible. PMID- 12547436 TI - High-level ab initio calculations on the energetics of low-lying spin states of biologically relevant transition metal complexes: a first progress report. AB - Although DFT is the unrivaled method of choice for quantum chemical studies of bioinorganic problems, little is known about its ability to predict the energetics of the low-lying electronic states of transition metal complexes. The first high-level ab initio calculations aimed at calibrating DFT vis-a-vis this issue indicate that, despite its many successes, DFT is far from infallible. In the short term, additional calibration of DFT against more elaborate ab initio methods remains an important goal for computational bioinorganic researchers. In the longer term, we are optimistic that high-level ab initio methods such as CASPT2 and CCSD(T) will be regularly used to study realistic molecules of genuine biochemical interest. PMID- 12547437 TI - Quantum chemical calculations of spectroscopic properties of metalloproteins and model compounds: EPR and Mossbauer properties. AB - Recently developed theoretical methods to predict EPR and Mossbauer parameters open the way for close interactions between theorists and experimentalists to elucidate the geometric and electronic structures of metalloenzymes and model complexes and to obtain insight into their reactive properties. Spectral calculations (g-values, hyperfine couplings, zero-field splittings, isomer shifts and quadrupole splittings) are also a means to validate theoretical approaches and therefore complement the prediction of geometries, reaction energies and transition states. PMID- 12547438 TI - Combined quantum and molecular mechanics calculations on metalloproteins. AB - The combination of quantum mechanics and molecular mechanics (QM/MM) methods is one of the most promising approaches to study the structure, function and properties of proteins. The number of QM/MM applications on metalloproteins is steadily increasing, especially studies with density functional methods on redox active metal centres. Recent developments include new parameterised methods to treat covalent bonds between the quantum and classical systems, methods to obtain free energy from QM/MM results, and the combination of quantum chemistry and protein crystallography. PMID- 12547439 TI - Molecular dynamics simulations of metalloproteins. AB - Molecular dynamics simulations are now commonly applied to metalloproteins, despite the challenges introduced by the presence of metal ions. Force field parameters are nowadays available also for these 'exotic' atoms and several biological systems have been successfully studied. Some of the most relevant results and methodological advancements are reviewed. PMID- 12547440 TI - Molecular modeling of ion channels: structural predictions. AB - Recent advances in membrane protein crystallography have greatly increased structural information of channels permeating metal ions. Structural bioinformatics techniques and molecular dynamics calculations are providing structural models of ion channels for which the three-dimensional structure is not known. Most of the reported structure prediction studies focus on K(+) channels and are based on the KcsA K(+) channel structure. PMID- 12547442 TI - Clinical epidemiology. what, who, and whither. AB - Clinical epidemiology, the what, was introduced by John Paul in 1938, as a new basic science for preventive medicine. Its definition subsequently took on a more bedside tone, but continues to be adapted to the needs of its practitioners. Clinical epidemiology, the who, centers on Alvan Feinstein and the way that he led the field and nurtured so many of its practitioners. Clinical epidemiology, the whither, describes its more recent development and its impact on five evolutions and revolutions: in evidence generation, its rapid critical appraisal, its efficient storage and retrieval, evidence-based medicine, and evidence synthesis. PMID- 12547443 TI - Feinstein and study design. AB - Dr. Alvan Feinstein saw himself as the father of "clinical epidemiology" in the modern meaning of this term, of this "new intellectual domain of modern medical science." In this role, he saw himself as drawing from his "clinical sophistication" and from "the rigorous scientific demands" to which "clinicians are accustomed," while "public health" epidemiologists "often use a more arbitrary set of standards." His conception of the scope of clinical epidemiology was remarkably Catholic and the same was the case in respect to cause-effect research in it. In the latter, he was firmly committed to the randomized-trial paradigm, including in his teachings on study design in etiologic research. Characteristically original, many of Dr. Feinstein's study-design ideas remain controversial. PMID- 12547444 TI - The apprenticeship of an editor and the secret role of a mentor. AB - Dr. Alvan R. Feinstein, late editor of the Journal of Clinical Epidemiology, is remembered by his fellow editor in this article. His role is highlighted as an innovative gatekeeper of clinical and epidemiological communications and as an effective secret mentor of his younger partner. The profile of his editorial practice is summarized as "six commandments" and the Golden Rule of pursuing attainable excellence. PMID- 12547445 TI - Alvan Feinstein and the art of consulting: how to define a research question. AB - When thinking of a new study, the most important task is to define its goals and to choose a design to match those goals. Alvan Feinstein described how he went about consulting others on defining a research question in his book, "Clinical Epidemiology, the Architecture of Clinical Research." In this paper, the author reminisces about how he learned and tried to apply those principles. PMID- 12547446 TI - Challenges and opportunities in evaluating diagnostic tests. AB - The field of clinical research conducted to evaluate diagnostic tests has evolved substantially over the last 25 years. This article discusses three current challenges and opportunities in conducting such research: the limitations of the "traditional focus" of clinical epidemiology in evaluating tests; how different decision makers use data produced by clinical epidemiology research, and what those uses mean for future research; and special challenges and opportunities in evaluating molecular-based tests. PMID- 12547447 TI - Evaluation of interventions. AB - The first modern randomized controlled trial was published in 1948, and featured randomly allocated treatment groups, blinded outcome assessment, and a sufficient number of patients. Randomized controlled trials are now accepted as the best possible way to assess the effects of clinical interventions, but good trials sometimes disagree. Although the elements of a credible trial have been defined, the relative importance of these elements to results is not known. Information from clinical trials may be difficult to use in the care of patients if trial outcomes are not patient centered or the conditions of the trial are very different from the situation in clinical practice. Observational studies are often, but not always, an adequate substitute from randomized trials. Systematic reviews with meta-analyses may reveal patterns of results when individual trials do not, and so complement the information content of large trials. When trials disagree, resolution may be found not by from more trials but also from critical appraisal of individual trials, weighing the totality of the evidence, and Bayesian reasoning. PMID- 12547448 TI - The false positive therapeutic trial. AB - I do not know how many randomized clinical trials are positive, and how many of these trials are false positive. Still, I get the feeling that among the published ones that are positive, the majority is false positive. This article tells you how you can end a trial with a positive result even if your trial data do not or hardly support your favorite treatment. It deals with many deliberate and less deliberate biases, in the introduction, in the patients and methods section, in the results, and in the data interpretation and report. PMID- 12547449 TI - Epidemiologic methods: the "art" in the state of the art. AB - When performing empirical research in public health and medicine, the investigator is typically faced with a variety of methodologic issues to resolve at the design and analysis stages of the research. The investigator must specify the research question, conceptualize and operationalize the variables to be measured, consider several research designs to choose from, determine appropriate measures of disease frequency and effect, address potential biases, devise the analytic strategies to follow, choose the appropriate software procedures or packages to use, carry out the analysis, and interpret the results. Most of these issues concern principles and methods of epidemiology and biostatistics, which, taken together, embody the methodologic "science" that underlies such research. Nevertheless, in bringing all these issues together to achieve a coherent and valid research conclusion, there is an "art" that is required that goes beyond the quantitative mechanics involved in carrying out the research. The art part is not easily quantifiable and typically is more skillfully performed as one becomes a more experienced investigator. Nevertheless, such methodologic art can be addressed methodologically through guidelines that identify the options or strategies available and suggest how appropriate choices can be made from such options or strategies. Selection bias and mathematical modeling are especially addressed. PMID- 12547450 TI - Between iatrotropic stimulus and interiatric referral: the domain of primary care research. AB - The "iatrotropic stimulus" is a typical example of Alvan Feinstein's terminology and his clinical way of thinking about medical research. Feinstein specified "interiatric referral" as an additional important factor affecting "the likelihood that patients will appear at the particular setting in which research is being conducted." He emphasized these phenomena as highly relevant for clinical judgment, clinical research, and clinical epidemiology. This has stimulated the realization of original clinical research in international primary medical care. It has helped to recognize the characteristics of the spectrum of health problem and morbidity patterns presented to primary care physicians and to acknowledge the position of generalistic primary care research between open population research and clinical research in referred (specialist) care. Accordingly, primary care-specific challenges in investigating (early) diagnosis, prognosis, treatment, and referral decisions have emerged. This research supports adequate decision-making by the primary care physician, which is important for effective health care. Although Feinstein was a clinical specialist and has always lived in a country without a very strong primary care tradition, he not only recognized the value of primary care research but also substantially contributed to its development. PMID- 12547451 TI - The controversial controversy of a passionate controversialist. AB - Alvan Feinstein was among the pioneers who perceived the need to apply epidemiologic principles and methods to clinical medicine. In his attempt to convert his peers to his own views, he was very liberal in criticizing the work of others. Epidemiologists still recall his 1988 article in Science entitled "Scientific Standards in Epidemiologic Studies of the Menace of Daily Life," in which he criticized observational epidemiology for its lack of scientific rigor. In this context, Feinstein's work has also been the center of controversies. One in particular, related to the fact that he carried out research and consulted for industries, in particular the tobacco industry, has tainted the last years of Feinstein's career. My purpose in this article is to discuss Feinstein's relationship with the tobacco industry. It is not to judge the validity of his work. Against this background, a question of historic relevance, one that can be discussed on the basis of some evidence, is whether Feinstein, world-reknowned epidemiologist and editor of this journal from 1982 until his recent death, helped the tobacco industry to publish articles minimizing the deleterious effects of smoking in the Journal of Chronic Diseases and (since 1988) the Journal of Clinical Epidemiology. To address this question I have searched the articles related to the effects of tobacco that were published in the Journal since 1978. The tobacco industry has had the opportunity, via its consultants, to defend its own ideas in the "Variance and Dissent" section. Perhaps in hindsight Feinstein could be criticized for not having clearly indicated the sponsorship of the tobacco industry behind these publications, of which he was fully aware. However, this does not suffice to infer that he was the tobacco industry's "man." Feinstein's attitude in matters of publication appears balanced. PMID- 12547452 TI - No creative medical science without humor. AB - In his time with the Journal of Clinical Epidemiology and before, Dr. Alvan Feinstein espoused a philosophy of the study of epidemiology as a beneficial, valid science. Not only was Feinstein somewhat of a nonconformist in the field of epidemiology, he had a self-denigrating sense of humor as well. This sense of humor can be showcased by taking a look at one of the articles he approved for publication, which could be retitled as "The Finster Saga." PMID- 12547465 TI - Abnormal prefrontal cortex function during response inhibition in Turner syndrome: functional magnetic resonance imaging evidence. AB - BACKGROUND: Turner syndrome (TuS) arises from the partial or complete absence of one X chromosome. Although neuropsychological studies report impaired attentional function and response inhibition in TuS, the neural correlates of these cognitive problems are unknown. METHODS: Eleven female subjects with TuS and 11 individually matched normal control subjects were imaged using functional magnetic resonance imaging while performing a Go/NoGo task. RESULTS: Groups did not differ on accuracy or reaction time; however, the TuS group activated more in the bilateral superior and middle frontal gyri than control subjects. Control subjects did not activate more than the TuS group in any region. CONCLUSION: These findings suggest that female subjects with TuS compensate for executive dysfunction via recruitment of additional prefrontal cortex regions involved in inhibition, attention, and working memory, functions necessary for successful performance of Go/NoGo tasks. Elucidating brain function in TuS will advance our understanding of the influence of X-chromosome genes on neurodevelopment and brain function and contribute to planning future intervention strategies. PMID- 12547466 TI - Atomoxetine in adults with ADHD: two randomized, placebo-controlled studies. AB - BACKGROUND: [corrected] Attention-deficit/hyperactivity disorder (ADHD) has been less studied in adults than in children, and the treatment studies reported to date have been small, single-center trials. To assess the efficacy of atomoxetine, a new and highly selective inhibitor of the norepinephrine transporter, we conducted two large, multicenter treatment trials. METHODS: Two identical studies using randomized, double-blind, placebo-controlled designs and a 10-week treatment period were conducted in adults with DSM-IV-defined ADHD as assessed by clinical history and confirmed by a structured interview (study I, n = 280; study II, n = 256). The primary outcome measure was a comparison of atomoxetine and placebo using repeated measures mixed model analysis of postbaseline values of the Conners' Adult ADHD Rating Scale. RESULTS: In each study, atomoxetine was statistically superior to placebo in reducing both inattentive and hyperactive and impulsive symptoms as assessed by primary and secondary measures. Discontinuations for adverse events among atomoxetine patients were under 10% in both studies. CONCLUSION: Atomoxetine appears to be an efficacious treatment for adult ADHD. Its lack of abuse potential may be an advantage for many patients. PMID- 12547467 TI - Reduced thalamic volume in high-functioning individuals with autism. AB - BACKGROUND: In this study, specific consideration is given to a role for the thalamus in autism. METHODS: A volumetric analysis of the thalamus was conducted using magnetic resonance imaging, based on segmentation of continuous 1.2 mm(3) coronal images. The sample consisted of 12 high-functioning individuals with autism, mean age of 21.0 years (SD = 10.4) and mean IQ of 106.4 (SD = 18.3). Normal control subjects were selected to match this group; the mean age was 18.1 years (SD = 6.3); mean IQ was 108.8 (SD = 15.6). RESULTS: Unadjusted mean thalamic volume was not significantly different; however, there were significant differences in the relationship between thalamic volume and total brain volume (TBV). The correlation was strong and positive in the control group but statistically nonsignificant in the autism group. Group differences were found when adjustments were made for TBV, achieved by grouping subjects' measurements on this variable using a split median procedure. Mean thalamic volume was significantly reduced in the autism group relative to normal control subjects, specifically within the high TBV group. CONCLUSION: The increase in thalamic volume with increase in TBV was not seen in autism, suggesting underdeveloped connections between cortical and subcortical regions and indicating a need to examine this structure further. PMID- 12547468 TI - Maternal smoking during pregnancy and risk to boys' conduct disturbance: an examination of the causal hypothesis. AB - BACKGROUND: We undertook this study to determine whether the widely replicated link between maternal smoking and conduct disturbance (Cd) is better explained by a model of direct causation or of mother-offspring transmission of a latent Cd variable. METHODS: Family data collected on 538 adolescent twin boys from the Virginia Twin Study of Adolescent Behavioral Development (VTSABD) was used to compare two alternative models: 1) a model composed of a latent transmissible factor that influences mother's juvenile conduct symptoms, smoking during pregnancy, and subsequent Cd and smoking in her adolescent boys; and 2) a model specifying a direct causal path from mother's smoking to child Cd. RESULTS: The maternal-offspring transmission model fit the data as well as a model specifying a direct causal path from maternal smoking to child Cd. CONCLUSIONS: Our results suggest that the association between maternal smoking during pregnancy and boys' Cd symptoms may be attributed to the transmission of a latent Cd factor and not to a direct effect of the smoking. Our results challenge previous findings of a direct effect of prenatal smoke exposure on risk to Cd once other etiologic factors are considered. PMID- 12547469 TI - A preliminary placebo-controlled trial of selegiline hydrochloride for smoking cessation. AB - BACKGROUND: Since dopaminergic mechanisms appear to be involved in nicotine dependence, we studied the safety and efficacy of the monoamine oxidase B inhibitor selegiline hydrochloride compared with placebo for smoking cessation in nicotine-dependent cigarette smokers. METHODS: Forty subjects with DSM-IV nicotine dependence were randomized to: 1) selegiline hydrochloride (5 mg p.o. twice daily) or 2) placebo in an 8-week trial. Outcome measures included smoking cessation rates, treatment retention, and medication side effects. RESULTS: Selegiline hydrochloride increased trial end point (week 8) 7-day point prevalence smoking cessation rates (selegiline hydrochloride, 9/20 [45.0%]; placebo, 3/20 [15.0%], odds ratio = 4.64, 95% CI, 1.02-21.00, p <.05), and smoking cessation rates during the last 4 weeks of the trial (selegiline hydrochloride, 6/20 [30.0%]; placebo, 1/20 [5.0%], odds ratio = 8.14, 95% CI, 0.88-75.48, p =.07) in comparison with placebo. Six-month follow-up 7-day point prevalence smoking cessation rates were reduced compared with trial end point (selegiline hydrochloride, 4/20 [20.0%]; placebo, 1/20 [5.0%], odds ratio = 4.75, 95% CI, 0.48-46.91, p =.18). Treatment retention was similar between drug and placebo groups (p =.13), and selegiline hydrochloride was well tolerated in cigarette smokers. CONCLUSION: This preliminary study suggests that selegiline (10 mg/day) is safe for use and enhances smoking cessation rates compared with placebo in nicotine-dependent cigarette smokers. PMID- 12547470 TI - Smaller orbital frontal cortex volumes associated with functional disability in depressed elders. AB - BACKGROUND: Depression is associated with significant functional impairment. Recent evidence has linked the orbital frontal cortex (OFC) with depression. We examined the relationship between OFC volumes in older subjects and impairment in the basic (BADL) and instrumental (IADL) activities of daily living. METHODS: The sample consisted of 81 subjects aged 60 years or older; 41 were depressed subjects and 40 healthy control subjects. In a structured interview, subjects reported their medical history and ability to perform both BADL and IADL. Subjects then had a brain magnetic resonance imaging (MRI) scan; the OFC was manually traced bilaterally using neuroanatomical landmarks. Logistic regression was used to examine the effect of OFC volume on BADL and IADL while controlling for the effects of total brain volume, subject status, medical comorbidity, and demographic factors. RESULTS: Smaller OFC volumes, along with greater cognitive impairment as measured by the Mini-Mental State Examination, were significantly associated with BADL impairment. Smaller OFC volumes and being depressed were significantly associated with IADL impairment. CONCLUSIONS: Smaller OFC volumes are independently associated with functional impairment, supporting its role in depression. Further studies are needed to determine how smaller OFC volumes are related to other MRI abnormalities associated with depression and functional impairment. PMID- 12547471 TI - Regional cerebral blood flow in depressed patients with white matter magnetic resonance hyperintensity. AB - BACKGROUND: Functional neuroimaging studies have consistently demonstrated decreased regional cerebral blood flow (rCBF) or metabolism in the frontal lobe, temporal lobe, or anterior cingulate gyrus of depressed patients. On the other hand, white matter hyperintensity as defined by magnetic resonance imaging (MRI) has been the most consistently replicated finding in structural neuroimaging studies on depression; however, these functional and structural neuroimaging findings of depression have not been well integrated. We aimed to clarify the possible associations of MRI-defined subcortical hyperintensities with rCBF changes in depressed patients. METHODS: Twelve depressed patients with subcortical hyperintensities defined by MRI, 11 depressed patients without MRI hyperintensities, and 25 healthy volunteers underwent 99mTc ECD SPECT. Group comparisons of their rCBF and correlation analysis between MRI hyperintensity and rCBF in patients were performed with a voxel-based analysis using statistical parametric mapping (SPM) software. RESULTS: Depressed patients showed decreased rCBF compared with control subjects in the frontal lobe, temporal lobe, and anterior cingulate gyrus whether subcortical hyperintensity existed or not; however, the patients with MRI hyperintensity showed decreased rCBF in the thalamus, basal ganglia, and brainstem in addition to cortical areas. Further, the score for white matter hyperintensity correlated negatively with rCBF in subcortical brain structures, including the thalamus and right basal ganglia. CONCLUSION: Our study indicates that depressed patients with MRI hyperintensities may have dysfunction in subcortical brain structures in addition to dysfunction in the fronto-temporal cortical structures. PMID- 12547472 TI - High numbers of circulating activated T cells and raised levels of serum IL-2 receptor in bipolar disorder. AB - BACKGROUND: Previously, we found an increased prevalence of thyroid autoantibodies in patients with bipolar disorder. In the present study, we investigated other signs of immune activation in bipolar patients, in particular an activation of the T cell system. METHODS: Fluorescence activated cell scanning (FACS) analysis was performed on lymphocytes of 64 outpatients with DSM-IV bipolar disorder using the T cell marker CD3 in combination with the activation markers MHC-class II, CD25, CD69 or CD71. In 34 patients, these assays were repeated after an interval of 2 years. In addition, T cell activation was determined by measuring serum soluble IL-2 receptor (sIL-2R) in 172 bipolar outpatients. Outcomes were compared with a healthy control group. RESULTS: Significantly higher numbers of circulating activated T cells and raised sIL-2R levels were found in euthymic, manic, and depressed bipolar patients when compared with healthy controls. In general, these abnormalities were stable over time. Manic patients showed significantly higher levels of sIL-2R in comparison with depressed patients. CONCLUSION: The T cell system was found to be activated in both symptomatic and euthymic patients with bipolar disorder. The pathophysiological significance of these findings remains to be explored. PMID- 12547473 TI - Norepinephrine-gamma-aminobutyric acid (GABA) interaction in limbic stress circuits: effects of reboxetine on GABAergic neurons. AB - BACKGROUND: Reboxetine is a selective norepinephrine (NE) reuptake inhibitor that exerts significant antidepressant action. The current study assessed norepinephrine-gamma-aminobutyric acid (GABA)-ergic mechanisms in reboxetine action, examining glutamic acid decarboxylase (GAD) mRNA expression in limbic neurocircuits following reboxetine within the context of chronic stress. METHODS: Male rats received 25 mg/kg reboxetine/day, p.o. Reboxetine and vehicle animals were exposed to 1 week of variable stress exposure or handling. Behavioral responses to stress (open field) were tested on day 7, and animals were killed on day 8 to assess neuroendocrine stress responses and limbic GAD65/67 mRNA regulation (in situ hybridization). RESULTS: Reboxetine significantly decreased behavioral reactivity in the open field. Reboxetine administration did not affect expression of GAD65/67 mRNA in handled rats; however, administration to stressed animals reduced GAD67 (but not GAD65) mRNA in the medial amygdaloid nucleus, posteromedial bed nucleus of the stria terminalis, and dentate gyrus. In contrast, GAD65 mRNA expression was increased by reboxetine in the lateral septum of stressed animals. CONCLUSIONS: Norepinephrine pathways appear to modulate synthesis of GABA in central limbic stress circuits. Decreases in GABA synthetic capacity suggest reduced activation of stress-excitatory pathways and enhanced activation of stress-inhibitory circuits, and is consistent with a role for GABA in the antidepressant efficacy of NE reuptake inhibitors. PMID- 12547474 TI - Changing cortical excitability with low-frequency transcranial magnetic stimulation can induce sustained disruption of tactile perception. AB - Transcranial magnetic stimulation (TMS) is promising as a therapeutic tool, and TMS of the motor system has served as a model for regionally specific modulations of cortical excitability. It is unclear, however, to what extent response characteristics of the motor cortex are representative of other brain systems. We wanted to determine whether TMS could induce a sustained disruption of somatosensory processing beyond the stimulation duration, similar to observations in the motor system. We applied 1-Hz TMS at 110% of subjects' motor thresholds for a variable duration over the right and left somatosensory cortex before subjects performed a tactile frequency discrimination task with the left hand. Tactile discrimination was impaired only after TMS over the right somatosensory cortex (analysis of variance: p <.01). The duration of this impairment correlated with the duration of the preceding TMS; the effect lasted approximately 2 min after 5 min of TMS, 4 min after 10 min of TMS, and 8 min after 20 min of TMS. Two conclusions arise: 1) low-frequency TMS can interfere with tactile perception in a robust and sustained way, and 2) TMS dosing parameters effective in the motor system are also effective in the somatosensory system and may reflect a modality independent response characteristic of the cerebral cortex. PMID- 12547475 TI - Low CSF somatostatin associated with response to nimodipine in patents with affective illness. AB - BACKGROUND: In patients with depression, treatment with nimodipine has been shown to increase cerebrospinal fluid (CSF) somatostatin (SRIF) and ameliorate baseline global cerebral hypometabolism. This study was conducted to assess whether a low baseline level of CSF SRIF was associated with response to nimodipine treatment. METHODS: Twenty-one depressed patients underwent lumbar puncture for analysis of CSF somatostatin-like immunoreactivity (SRIF-LI) during a medication-free period and after at least 6 weeks of nimodipine monotherapy. Twenty-five healthy control subjects were utilized as a comparison group. Clinical improvement was assessed using the Clinical Global Impression Scale for Bipolar Illness. RESULTS: As predicted, baseline CSF SRIF-LI was significantly lower in eventual nimodipine responders (33.1 +/- 2.8 pg/mol) compared to eventual nonresponders [41.9 +/- 2.6 pg/mL; t(19) = 1.98, p =.03, one-tailed]. CONCLUSIONS: Low baseline CSF somatostatin in depression may be associated with response to nimodipine, which in turn may be related to the ability of nimodipine to increase CSF somatostatin. PMID- 12547476 TI - Proton spectroscopy in medication-free pediatric attention-deficit/hyperactivity disorder. AB - BACKGROUND: The frontal-striatal pathway has been previously implicated in the neuropathology of attention-deficit/hyperactivity disorder (ADHD). Hence, we used proton magnetic resonance spectroscopy (1H-MRS) to examine metabolite levels in the prefrontal cortex of children with ADHD. METHODS: Nine age- and gender matched case-control pairs were examined, ages 7 to 16 years. A long-echo 1H-MRS scan was acquired from the right prefrontal cortex and left striatum in all subjects. Compounds that can be visualized with 1H-MRS include N-acetyl-aspartate (NAA), glutamate/glutamine/gamma-aminobutyric acid (Glx), creatine/phosphocreatine (Cr), and choline compounds (Cho). RESULTS: Frontal striatal glutamatergic resonances were elevated in the children with ADHD as compared to healthy control subjects. No differences were noted in NAA, Cho, or Cr metabolite ratios. CONCLUSIONS: These findings suggest that frontal-striatal Glx resonances may be increased in children with ADHD in comparison with healthy control subjects. PMID- 12547477 TI - Mirtazapine vs. placebo in posttraumatic stress disorder: a pilot trial. AB - BACKGROUND: Based on an earlier pilot study, as well as a theoretical consideration of its mechanism of action, we undertook a placebo-controlled, double-blind trial of mirtazapine in posttraumatic stress disorder. METHODS: Twenty-nine patients were randomized to receive drug up to 45 mg/day or placebo double-blind on a 2:1 ratio for 8 weeks, with data being available for analysis in 26. Primary outcome measures comprised the Short Posttraumatic Stress Disorder Rating Interview (SPRINT) Global Improvement item and total score. Secondary measures comprised the Davidson Trauma Scale, Structured Interview for Posttraumatic Stress Disorder and Hospital Anxiety Depression Scale. Adverse events were also measured. RESULTS: On the Short Posttraumatic Stress Disorder Rating Interview Global Improvement measure, rates of response were 64.7% and 20.0% for mirtazapine and placebo. Treatment effects in favor of mirtazapine were noted on the Short Posttraumatic Stress Disorder Rating Interview global, Structured Interview for Posttraumatic Stress Disorder, and Hospital Anxiety Depression Scale anxiety subscale scores. The drug was well tolerated. CONCLUSIONS: Mirtazapine was more effective than placebo on some measures in posttraumatic stress disorder and general anxiety symptoms. PMID- 12547479 TI - Epidemiologic studies of occupational pesticide exposure and cancer: regulatory risk assessments and biologic plausibility. AB - Epidemiologic studies frequently show associations between self-reported use of specific pesticides and human cancers. These findings have engendered debate largely on methodologic grounds. However, biologic plausibility is a more fundamental issue that has received only superficial attention. The purpose of this commentary is to review briefly the toxicology and exposure data that are developed as part of the pesticide regulatory process and to discuss the applicability of this data to epidemiologic research. The authors also provide a generic example of how worker pesticide exposures might be estimated and compared to relevant toxicologic dose levels. This example provides guidance for better characterization of exposure and for consideration of biologic plausibility in epidemiologic studies of pesticides. PMID- 12547480 TI - Unemployment and the detection of early stage breast tumors among African Americans and non-Hispanic whites. AB - PURPOSE: To test the hypothesis that high unemployment predicts reduced detection of local breast tumors among African American and non-Hispanic white women in the Detroit, Michigan and Atlanta, Georgia SEER catchment areas. METHODS: We test the hypothesis with data for the 156 months from January 1985 through December 1997. RESULTS: In situ and local breast tumors in African American and non-Hispanic white women were less likely to be detected during periods of high unemployment. CONCLUSIONS: Contracting labor markets may impede women with symptoms from getting proper medical attention or distract women from discovering symptoms they would otherwise detect. African American women appear at greatest risk of having a tumor going undetected by virtue of labor market performance. PMID- 12547481 TI - Polybrominated biphenyl exposure and benign breast disease in a cohort of US women. AB - PURPOSE: We examined the relation between serum polybrominated biphenyl (PBB) levels and the risk of benign breast disease in a cohort of Michigan women unintentionally exposed to PBBs in 1973 and interviewed in 1997. METHODS: We used extended Cox models to generate adjusted hazard ratios; models included polychlorinated biphenyls (PCBs) and risk factors for benign breast disease reported in the literature. RESULTS: Two hundred fourteen (23%) of 951 women reported benign breast disease diagnosed by a physician. Compared with women with low PBB exposure, benign breast disease was not reported more frequently among those with moderate (>1-12 parts per billion [ppb]), (odds ratio [OR] 1.08, 95% confidence interval [CI] = 0.80-1.45), or high (>12 ppb), (OR 0.79, 95% CI = 0.46 1.38) PBB exposure. PCB exposure was also not associated with self-reported physician-diagnosed benign breast disease. Age, smoking, and annual number of health-care provider visits were significantly associated with benign breast disease. CONCLUSIONS: Our analysis did not demonstrate an association between serum PBB level and self- reported physician-diagnosed benign breast disease. We did observe an increased risk of benign breast disease for women who smoked, an association that has not been consistently found in previous studies. PMID- 12547482 TI - Dietary factors and the risk of gastric cancer among Japanese women: a comparison between the differentiated and non-differentiated subtypes. AB - PURPOSE: Using data from the Hospital-based Epidemiologic Research Program at Aichi Cancer Center (HERPACC), a case-referent study was performed to clarify whether dietary habits differentially impact on the risk of female gastric cancers of different histological subtypes. METHODS: The study subjects comprised 508 histologically confirmed female gastric cancer cases (156 differentiated, 352 non-differentiated), identified via hospital cancer registry and surgical records between 1988 to 1998. The referents were 36,490 cancer-free first-visit female outpatients over 30 years old presenting at the center within the same period of time. The odds ratios were estimated by logistic regression analyses and adjusted for potential confounding factors. RESULTS: Frequent vegetable and fruit conferred reduced risk of both differentiated and non-differentiated female gastric cancers in similar patterns. In addition, reduced risk was observed for intake of fish and soybean products, particularly for the non-differentiated type. CONCLUSIONS: These results suggest that both differentiated and non differentiated female gastric cancers are preventable by frequent intake of vegetable and fruit, fish and soybean products, suggested to be common protective factors, although the possibility of some variation in the impact of the diet on different histopathological entities needs to be further addressed. PMID- 12547484 TI - Breast cancer surveillance using gridded population units, Connecticut, 1992 to 1995. AB - PURPOSE: To assess geographic variation in invasive breast cancer across Connecticut using gridded population areas to enumerate cases and the population at-risk. METHODS: The state's land mass was divided into 5168, 1-by-1 square mile areas and the population of women, 20+ years of age, within each location was estimated by areal interpolation of the 1990 US Census Block Group STF-3A data file. Using information on breast cancer incidence, 1992 to 1995, from the Connecticut Tumor Registry, latitude-longitude coordinates for place of residence at the time of breast cancer diagnosis were determined for 8530 records and assigned to appropriate grid locations. A spatial scan statistic was used to detect variation in incidence and test the significance of observed differences across the state. Standardized Incidence Ratios (SIRs) described the proportional change in the age-adjusted breast cancer incidence rate across gridded locations. RESULTS: The statewide age-adjusted invasive cancer incidence rate was 163.6/100,000 women/year. The spatial scan statistic identified three locations around Connecticut with significantly low incidence rates and four places where rates were significantly high. The most probable place of low incidence was rural Northeastern Connecticut where risk of disease, relative to elsewhere around the state, was 0.73 (p = 0.001). The most probable location of elevated incidence was a suburban location in Southwestern Connecticut with a relative risk of 2.02 (p = 0.001). CONCLUSIONS: Visual representation of disease incidence and underlying populations at-risk according to gridded units provides a useful tool for assessing small area variation in disease patterns. PMID- 12547483 TI - An international comparison of breast cancer survival: Winnipeg, Manitoba and Des Moines, Iowa, metropolitan areas. AB - PURPOSE: Extending previous Canadian-United States cancer survival comparisons in large metropolitan areas, this study compares breast cancer survival in smaller metropolitan areas: Winnipeg, Manitoba and Des Moines, Iowa. METHODS: Manitoba and Iowa cancer registries, respectively, provided a total of 2,383 and 1,545 women with breast cancer (1984 to 1992, followed until December 31, 1997). Socioeconomic data for each person's residence at the time of diagnosis was taken from population censuses. RESULTS: Socioeconomic status and breast cancer survival were directly associated in the US cohort, but not in the Canadian cohort. Compared with similar patients in Des Moines, residents of the lowest fifth of income areas in Winnipeg experienced a significant 5-year survival advantage (survival rate ratio [SRR] = 1.14). In these lowest income areas, the Canadian survival advantage was larger among women aged 25 to 64 years (SRR = 1.23), and this was observed in the middle fifth of income areas among this younger cohort (SRR = 1.11). The Canadian survival advantage even seemed apparent in the poorest neighborhoods with relatively high representations of Aboriginal people (SRR = 1.16). CONCLUSION: This study replicated the finding of advantaged Canadian cancer survival in smaller metropolitan areas that had been consistently observed in larger metropolitan areas. Canada's single payer health care system seems to offer similar advantages across a number of diverse urban contexts. PMID- 12547485 TI - Extended mortality follow-up of a cohort of dry cleaners. AB - PURPOSE: The mortality follow-up of a cohort of dry cleaners was extended to further evaluate cancers risks associated with organic solvents. METHODS: The underlying and contributing causes of death among 5,369 members of a dry cleaning union in St. Louis were determined through December 31, 1993. The mortality experience of the cohort was compared to that of the US population adjusted for age at entry, year of death, race and gender. RESULTS: The total mortality was about as expected (SMR = 1.0, N = 2351, 95% CI = 1.0-1.1). Excesses were observed for emphysema (SMR = 1.7, N = 21, 95% CI = 1.0-2.5), Hodgkin's disease (SMR = 2.0, N = 5, 95% CI = 0.6-4.6) and cancers of the esophagus (SMR = 2.2, N = 26, 95% CI = 1.5-3.3), larynx (SMR = 1.7, N = 6, 95% CI = 0.6-3.7), lung (SMR = 1.4, N = 125), 95% CI = 1.1-1.6), and cervix (SMR = 1.6, N = 27, 95% CI = 1.0-2.3). These excesses occurred among men and women and blacks and whites. Bladder cancer was elevated among white men and women and kidney cancer among black men and women, but not significantly so. None of these causes of death showed strong relationships with duration or estimated level of exposure to dry cleaning solvents, although relative risks for cancers of the larynx, lung and kidney were larger among subjects estimated to have higher levels of exposure and risks from bladder cancer and chronic nephritis were greater among persons who entered the union after 1960. CONCLUSION: The excesses observed are unlikely to be due to chance because most occurred in earlier as well as the recent follow-up. The specific factors contributing the excesses, however, are not clear. Socioeconomic, lifestyle, and occupational exposures are all possibilities. Lack of information on socioeconomic and lifestyle factors hampers evaluation. PMID- 12547486 TI - Angiotensin I-converting enzyme (ACE) gene polymorphism in relation to physical performance, cognition and survival--a follow-up study of elderly Danish twins. AB - PURPOSE: Studies of younger individuals have suggested an association between ACE genotype and physical and cognitive performance. Using a longitudinal study of elderly twins we studied the association between ACE genotype and physical and cognitive functioning and survival in old age. METHODS: Participants were 684 twins aged 73+ years from the 1997 and 1999 surveys of the Longitudinal Study of Aging Danish Twins. Cognitive skills were assessed by the MMSE, while physical abilities were determined through self-report in 1997 and through both self report and measurement of performance in two physical tasks in 1999. Survival status was obtained through linkage with a national death register. RESULTS: Neither physical nor cognitive performance was associated with ACE genotype at baseline in 1997, or at follow-up in 1999. For participants in both surveys longitudinal changes in these skills did not depend on ACE genotype. The relative risk of dying was increased in II compared with the DI and DD genotype with relative risks of 1.6 (95 percent confidence intervals 1.1-2.5) and 1.3 (0.8 2.1), respectively. CONCLUSION: We found no substantial effects of ACE genotype on physical and cognitive performance, or rate of change among elderly. Persons with the D allele may have a lower mortality at older ages. PMID- 12547487 TI - Occupant- and crash-related factors associated with the risk of whiplash injury. AB - PURPOSE: Given that a motor vehicle crash (MVC) had occurred, to evaluate whether occupant- and crash-related factors, such as age, gender, seating position and type of MVC are associated with the risk of whiplash injury. METHODS: A study of occupants in cars covered by motor insurance at one of the largest insurance companies in Sweden, was undertaken during a one-year period. The study population comprised all occupants in cars exposed to an MVC in which at least one occupant was injured (n = 7120). Adjusted estimates of the relative risk of whiplash injury, associated with the different factors, were calculated by means of binomial regression analysis. RESULTS: Considering different MVCs, rear-end collisions were associated with the highest relative risk of whiplash injury when compared with side impacts (1.82; 95% CI 1.68-1.96), while drivers showed the strongest association with respect to seating position when compared with passengers in the rear seat (1.78; 95% CI 1.60-1.97). Females had a somewhat higher relative risk of whiplash injury than males (1.20; 95% CI 1.16-1.25). Regarding age, the relative risk was moderately increased across the different age groups when compared with the oldest age group. No interaction was observed on the additive scale. CONCLUSIONS: Given that an MVC had occurred, subjects exposed to a rear-end collision and drivers had a substantial increased risk of whiplash injury, while age and gender were of minor importance. PMID- 12547488 TI - Regarding: a cohort study of systemic and local complications following implantation of testicular prostheses. PMID- 12547489 TI - Response to letter to the editor. regarding "The completeness, validity, and timeliness of AIDS surveillance data". PMID- 12547490 TI - Lower body anatomical distribution of solar ultraviolet radiation on the human form in standing and sitting postures. AB - Humans undertake their daily activities in a number of different postures. This paper aims to compare the anatomical distribution of the solar erythemal UV to human legs for standing and sitting postures. The exposure ratios to the legs (ratio of the UV exposure to a particular anatomical site compared to the ambient) have been measured with UV dosimeters for standing and sitting postures of a manikin. The exposure ratios for the legs ranged from 0 to 0.75 for the different anatomical sites for the sitting posture in summer (December through February) compared to 0.14 to 0.39 for the standing posture. In winter (June through August) the exposure ratios ranged from 0.01 to 0.91 for sitting to 0.17 to 0.81 for standing. For the anterior thigh and shin, the erythemal UV exposures increased by a factor of approximately 3 for sitting compared to standing postures. The exposure ratios to specific anatomical sites have been multiplied by the ambient erythemal UV exposures for each day to calculate the annual exposures. The annual erythemal exposures to the anterior thigh and ankle were predicted to be higher than 800 MED for humans sitting outdoors each day between noon and 13:00 h Australian Eastern Standard Time (EST). For humans standing outdoors during this time, the annual erythemal UV exposure averaged over each leg site was 436 MED, whereas, the averaged annual erythemal UV exposure was 512 MED for the sitting posture. Similarly, the annual erythemal UV exposure averaged over each of the sites was 173 MED for humans standing outdoors between 09:00 h EST and noon each Saturday morning and 205 MED for humans sitting outdoors during this time. These results show that there is increased risk of non-melanoma skin cancer and malignant melanoma to the lower body if no UV preventative strategies are employed while in a sitting posture compared to a standing posture. PMID- 12547491 TI - UV-B induced keratinocyte apoptosis is blocked by 2-selenium-bridged beta cyclodextrin, a GPX mimic. AB - Cell proliferation and cell death of keratinocytes are tightly regulated to ensure epidermal homeostasis. UV-B induces keratinocyte apoptosis. UV-B also induces lipid peroxidation of keratinocytes to increase their amount of malondialdehyde (MDA). These phenomena can be explained by the production of reactive oxygen species (ROS) induced by UV-B radiation. We synthesized 2 selenium-bridged beta-cyclodextrin (2-SeCD) to imitate glutathione peroxidase (GPX), an important antioxidant and established a damage system, in which keratinocytes can be damaged by Ultraviolet B (UV-B) radiation. Using this damage system we studied 2-SeCD protection of keratinocytes against injury induced by UV B. Experimental results showed that 2-SeCD could protect keratinocytes from apoptosis. Moreover, 2-SeCD inhibits lipid peroxidation of keratinocytes and scavenges ROS. 2-SeCD inhibits the UV-B induced apoptotic signal transduction. This antiapoptotic mechanism may be partly related to the elimination of hydrogen peroxide. PMID- 12547492 TI - Spectral UV in public shade settings. AB - The protective nature of specific shade environments was investigated by measuring the spectral UV in the shade for the three planes (horizontal, 45 degrees and vertical) and comparing this to that on a horizontal plane in full sun. Spectral UV irradiance measurements were made under clear sky conditions at a sub-tropical southern hemisphere site. The solar UV in the shade of a shade umbrella, covered verandah, covered sand pit and covered walkway were measured for an increasing solar zenith angle, between March and August, for the times of 11:30 a.m.-12:30 p.m. and 2:30-3:30 p.m. The ultraviolet protection factors provided ranged from 1.4 to 10. This research shows that there is sufficient UV in the shade to cause erythema on the human body in a short period of time. For the shade umbrella placed on dry grass the time able to be spent in the shade in the middle of the day before experiencing mild erythema increased from 35 to 60 min as the solar zenith angle increased from 33 to 52 degrees. Erythemal UV levels in the shade of a northern facing covered verandah, with trees in close proximity, were approximately up to five times less than the erythemal UV beneath the shade umbrella that had no surrounding trees. Shade structures must be given careful consideration when construction occurs. Even though the UV transmission through the materials may be very low, it is the construction of the entire shade setting that determines the exposure beneath the shade structure. PMID- 12547493 TI - Effects of short-term irradiation on photoinhibition and accumulation of mycosporine-like amino acids in sun and shade species of the red algal genus Porphyra. AB - The effect of irradiance (40 and 840 micromol photons m(-2) s(-1)) of short-term (48 h) irradiation on photosynthetic activity (estimated as oxygen evolution and as chlorophyll fluorescence), specific absorption and fluorescence excitation spectra, photosynthetic pigment accumulation (chlorophyll a and biliproteins) and UV-absorbing compounds (mycosporine-like amino acids, MAAs) was investigated in sun and shade species of the red algal genus Porphyra collected in Trondheimsfjord (Norway). In the sun type, high irradiance exposure (840 micromol photons m(-2) s(-1)) did not alter the Chl a concentration, however, exposure to a lower irradiance (40 micromol photons m(-2) s(-1)) for 48 h significantly increased the chlorophyll concentration. The content of MAAs was significantly higher in the suntype than in the shade type algae. Porphyra-334 is the main MAA in this species followed by shinorine. The total content of MAAs significantly (P<0.05) increased in the sun type after 48 h exposure to both high and low irradiances. However, in the shade type, porphyra-334 significantly decreased (P<0.05) after both high and low irradiance exposure. Photosynthetic activity (as oxygen evolution) and the optimal quantum yield (F(v)/F(m)), as an indicator of photoinhibition, decreased under low and high irradiance in the shade type algae and no full recovery was observed when the algae were transferred to very low irradiation. The sun type algae presented a higher capacity of acclimation to increased irradiance than the shade type algae. This high acclimation of sun type algae to short term high irradiance exposure (48 h) is explained by the higher thermal dissipation. This was estimated as the ratio of nonphotochemical quenching related to the light dose (q(N):dose) and by the accumulation of MAAs. PMID- 12547494 TI - Early reactions of light-induced protochlorophyllide and chlorophyllide transformations analyzed in vivo at room temperature with a diode array spectrofluorometer. AB - The steps of protochlorophyllide (Pchlide) photoreduction and subsequent chlorophyllide (Chlide) transformations which occur in the seconds to minutes time-scale were studied using a diode array spectrofluorometer in dark-grown barley leaves. The intensity of the excitation light was varied between 3 and 2,500 micromol m(-2) s(-1) and a series of fluorescence spectra were recorded at room temperature in the seconds and minutes time scales. In certain experiments, 77-K emission spectra were measured with the same equipment. The high quality of the spectra allowed us to run spectral resolution studies which proved the occurrence, at room temperature, of multiple Pchlide and Chlide forms found previously in 77-K spectra. The comparison of the 77-K and room-temperature spectra showed that the fluorescence yields of the nonphotoactive 633-nm Pchlide form and of the Chlide product emitting at 678 nm were temperature independent. The fluorescence intensity of aggregated NADPH-pigment-POR complexes (photoactive 656-nm Pchlide and 693-nm Chlide forms) were strongly increased at 77 K, while that of the NADP(+)-Chlide-POR (684-686-nm Chlide form) was much less affected by temperature. Information was obtained also about the dynamics of the transformation of pigment forms in the light at different photon densities. At low light intensities, the phototransformation of the 642-644-nm Pchlide form was faster than that of the 654-656-nm form. The relative amplitudes of Gaussian components related to different Chlide forms found after exposure to a constant amount of photons strongly depended on the light intensity used. Strong quenching of all Chlide components occurred upon prolonged exposure to high intensity light. These effects are discussed by considering the interconversion processes between different forms of the pigment-protein complexes, their relative fluorescence yields and energy migration processes. PMID- 12547495 TI - Different kinetics of photoinactivation of photosystem I-mediated electron transport and P700 in isolated thylakoid membranes. AB - Photoinactivation kinetics of photosystem I (PSI)-mediated electron transport rate was compared to that of P700 content at room (22 degrees C) and low (4 degrees C) temperatures in isolated spinach thylakoid membranes. The high light treatment was carried out under aerobic and anaerobic conditions. At 22 degrees C the decrease of electron transport rate showed first order exponential kinetics. The amount of P700 decreased linearly, being less affected in the first hours of illumination. During photoinhibition at 4 degrees C in the presence of oxygen, the kinetics of inactivation of PSI photochemical activity and the content of P700 were different. It was found that 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) had different protective effect on the electron transport rate and on P700 content at both temperatures. Treatment with high light intensity under N(2) atmosphere had no effect on the electron transport rate or P700 content. The possible degradation of PSI reaction centre proteins was determined using immunoblot methods. In the presence of linear electron transport at 22 degrees C correlation between formation of toxic hydroxyl radicals and inhibition of oxygen uptake was observed. PMID- 12547496 TI - Dimers of polyene antibiotic amphotericin B detected by means of fluorescence spectroscopy: molecular organization in solution and in lipid membranes. AB - Fluorescence emission from amphotericin B dissolved in 2-propanol-water was recorded in the spectral region 500-650 nm. The fluorescence excitation spectrum corresponds to the absorption spectrum of the monomeric drug. The large energy shift between the excitation and emission bands indicates that emission takes place from an energy level different than that responsible for absorption. These levels were attributed to the 2(1)A(g) and 1(1)B(u) states, respectively. Excitation of the same sample with short wavelength radiation (below 350 nm) yields light emission between 400 and 550 nm. The fluorescence excitation spectrum corresponding to this emission band displays distinct maxima at 350, 334 and 318 nm. This band was analyzed in terms of the exciton splitting theory and assigned to amphotericin B in a dimeric form, in which chromophores are spaced by 4.9 A. The binding energy of the dimers, determined to be 4.9 kJ/mol, indicates that the structures are stabilized by van der Waals interactions. The same type of molecular structures was also detected in the lipid membranes formed with dipalmitoylphosphatidylcholine. Linear dichroism of amphotericin B embedded in lipid multibilayers indicates that molecules are distributed between two fractions: parallel (38%) and perpendicular (62%) with respect to the membrane. The biological importance of such membrane organization is discussed. PMID- 12547497 TI - A rapid method for determining recent sunscreen use in field studies. AB - The role of sunscreens in preventing skin cancer and melanoma is the focus of ongoing research. Currently, there is no objective measure which can be used in field studies to determine whether a person has applied sunscreen to their skin, and researchers must use indirect assessments such as questionnaires. We sought to develop a rapid, non-invasive method for identifying sunscreen on the skin for use in epidemiological studies. Our basic method is to swab the skin, elute any residues which have been adsorbed onto the swab by rinsing in ethanol, and submit the eluted washings for spectrophotometric analysis. In a controlled study, we applied 0.1 ml of sunscreen to a 50 cm(2) grid on both forearms of 21 volunteers. Each forearm was allocated one of 10 different sunscreen brands. The skin was swabbed after intervals of 20 min, 1 h, 2 h and 4 h. In a field study conducted among 12 children aged 2-4 years attending a child care centre, sunscreen was applied to the faces of half the children. Swabs were then taken from the face and back of all children without knowledge of sunscreen status. In the controlled study, sunscreen was clearly detectable up to 2 h after application for all brands containing organic sunscreen, and marginally detectable at 4 h. In the field study, this method correctly identified all children with and without sunscreen. We conclude that spectrophotometric analysis of skin swabs can reliably detect the presence of sunscreen on the skin for up to 2 h after application. PMID- 12547498 TI - Will the idiotypic network help to solve natural tolerance? AB - A recent paper on immunity and tolerance to antigenic determinants on antibody variable-regions (idiotypes) brings back to light the potential of idiotypic interactions among antibodies, B cells and T cells in the regulation of specific immune activities. The functional significance of idiotypic regulation has been established in a variety of systems, both concerning the establishment of 'pre immune' diversity repertoires as well as the clonal regulation of immune responses to foreign and self-antigens. Also, recently the requirement for 'dominant' regulatory mechanisms in natural tolerance has received increasing support. It might thus be fruitful to evaluate the possibility that an idiotypic network has a fundamental role in the operation of the regulatory T cells that establish and maintain self-tolerance. PMID- 12547499 TI - Novel functions of the polymeric Ig receptor: well beyond transport of immunoglobulins. AB - The polymeric Ig receptor (pIgR) ensures efficient secretion of polymeric IgA (pIgA) at mucosal surfaces. On basal to apical transport across epithelial cells, the pIgR extracellular domain is cleaved, releasing secretory component (SC) in association with pIgA. This finds its raison d'etre in the recent observation that SC is directly involved in the protective function of secretory IgA. In addition, free SC exhibits scavenger properties with respect to enteric pathogens. However, although pIgR dedicates its life to mucosal protection, it also seems to permit pathogen entrance through the epithelial barrier. The multiple mechanisms that they are involved in make pIgR and SC instrumental to mucosal immunity. PMID- 12547500 TI - Determinant spreading and tumor responses after peptide-based cancer immunotherapy. AB - Modern immunological assays are very sensitive for detection of antigen-specific T cells. These assays are used to detect increased levels of T cells after peptide-based immunotherapy for cancer in an attempt to describe surrogate endpoints correlated with anti-tumor activity. Recent reports demonstrate that determinant spreading develops in a high frequency of subjects with tumor regression responses after this type of immunotherapy and could be valuable for trial monitoring and the design of more effective vaccines. PMID- 12547501 TI - Cancer immunoprevention: tracking down persistent tumor antigens. AB - The immune response can effectively hamper the progression of preclinical stages of tumor growth. Medicine in the postgenomic era offers an increasing possibility of detecting healthy individuals at risk of developing cancer who could benefit from tumor-preventive vaccines. The identification of novel tumor antigens that fulfill two conditions will be crucial for the development of cancer immunoprevention. First, an ideal antigen should have a crucial pathogenetic role in tumor growth to avoid the selection of antigen-loss variants. Second, the antigen should be recognizable by the immune system even in MHC-loss variants and should therefore be recognized both by antibodies and T cells. Identifying such antigens will also provide new targets for cancer immunotherapy. PMID- 12547502 TI - Will we be able to 'spot' an effective HIV-1 vaccine? AB - Many HIV-1 vaccine efforts have heavily emphasized class I-restricted CD8(+) cytotoxic T lymphocytes (CTLs) as a potentially important arm of immunity. Commonly used CTL assays describe only specificity and frequency, and not antiviral effects. However, increasing evidence indicates that CTL antiviral function is determined by the complex interplay of multiple virologic and cellular factors, and thus these measurements are an incomplete reflection of CTL efficacy. Our current understanding of the factors determining HIV-1-specific CTL antiviral efficacy is inadequate to interpret fully the significance of detected CTLs in vaccine and pathogenesis studies. To assess CTLs as a protective immune response, it will be crucial to elucidate these mechanisms and/or devise new assays that directly reflect the interaction of CTLs and HIV-1. PMID- 12547503 TI - Ankylosing spondylitis: a beta2m-deposition disease? AB - To explain the strong association between HLA-B27 and ankylosing spondylitis, we suggest that the release of beta(2)-microglobulin (beta(2)m) from a subpopulation of cell surface-expressed HLA-B27 molecules leads to beta(2)m-deposition within synovia and to the initiation of an inflammatory process, which culminates in destructive spondyloarthropathy. PMID- 12547504 TI - A central role for peripheral dendritic cells in the induction of acquired thymic tolerance. AB - Despite extensive negative selection in the thymus, numerous clones of self reactive T cells are normally exported to the periphery. In most instances, autoimmunity is prevented by regulatory T (Tr) cells, many of which are also of recent thymic origin. We have demonstrated recently that natural killer (NK) Tr thymocytes (THYr) can be induced by the injection of antigen into the eye, an immunologically privileged site; and that the intravenous infusion of antigen presenting cells (APCs) from such animals also induces NKT THYr. Furthermore, we have also observed that some of these APCs migrate to the thymus as CD11c(+) dendritic cells (DCs). Other authors have correlated the migration of DCs to the thymus with the generation of CD4(+)CD25(+) THYr. We therefore propose a novel tolerance induction pathway by which tolerogenic DCs routinely transport antigen (both self and nonself) from the periphery to the thymus, where they positively select THYr. We also propose that the ability of tolerogenic DCs to induce acquired thymic tolerance on demand might have important implications for the immunotherapy of autoimmunity and allotransplantation. PMID- 12547505 TI - HLA-G and MIC expression in tumors and their role in anti-tumor immunity. AB - Non-classical MHC class Ib molecules have attracted growing interest in recent years, especially because they interact with non-T-cell inhibitory or triggering receptors expressed on natural killer (NK) and T cells, suggesting that they have a role in immune recognition. Abnormalities in MHC class Ib expression are frequently found in human tumors of various histologies and might be associated with poor clinical outcome despite the local accumulation of immune competent cells. Available data suggest that the balance between activating and suppressing signals significantly influences the efficacy of the immune response and consequently of tumor progression. PMID- 12547506 TI - MHC specificity of iIELs. AB - Intestinal intra-epithelial lymphocytes (iIELs) are a major lymphocyte population, reside in close proximity to the intestinal lumen and are conserved throughout vertebrate evolution. iIELs consist of several unique T-cell phenotypes and express both non-rearranged innate immune receptors and rearranged adaptive immune receptors. The ligands for the innate immune receptors on iIELs, such as NKG2D (natural killer-cell receptor), often bind to non-classical MHC class I molecules, such as the human MHC class I-related molecules MICA or MICB. These ligands costimulate T-cell receptor (TCR)-mediated signaling. In most cases, the MHC molecules that bind to the TCR are still unknown. However, recent efforts to understand the MHC molecules that are involved in the development of and antigen recognition by iIELs have revealed several important results. Here, we focus systematically on recent developments in innate immunity and in TCR recognition of different subtypes of iIELs by various MHC molecules. PMID- 12547507 TI - Neutralizing antibodies in gene-defective hosts. AB - In a host with a normal immune system and a complete gene defect, the nondefective gene product will be immunogenic. Consequently, neutralizing antibodies against the respective protein can arise either 'spontaneously' or after immunization, as shown in patients and in animal models, such as knockout mice. Accordingly, patients with X-linked or homozygous autosomal gene defects are at risk of developing neutralizing antibodies, in particular after protein substitution or gene therapy. This Review compares and exemplifies the various genetic and immunological contexts that lead to 'neutralizing and generated by gene defect' or 'nagged' antibodies, and outlines implications and solutions for therapeutic strategies. PMID- 12547508 TI - How many lethal alleles? AB - Knowledge of the frequency of lethal mutant alleles in a population is important for our understanding of population genetics and evolution, and yet there have been few attempts to measure their number in wild populations. A new study has revealed unexpectedly low numbers of segregating lethal alleles in two species of fish. More experiments are needed, however, to know whether this result is general. PMID- 12547509 TI - Searching for nuclear-mitochondrial genes. AB - Recently, a novel strategy has been developed to identify yeast genes that are important for mitochondrial respiratory chain function. This approach found a large number of genes that were not previously thought to be involved, providing new candidate disease genes for mitochondrial disorders. These genes could cast light on the intricate relationship between genotype and phenotype in a wide range of inherited human diseases. PMID- 12547510 TI - Induced gene expression in human brain after the split from chimpanzee. AB - Despite only approximately 1% difference in genomic DNA sequence, humans and chimpanzees differ considerably in mental and linguistic capabilities, and in susceptibility to some diseases. A recent comparison of gene expression in human and great apes cast some light on the genetic basis of these differences, but more rigorous study is required. Our statistical reanalysis of these microarray data shows that there have indeed been dramatic alterations in the expression of genes in the human brain since the split from chimpanzees, mainly caused by a set of genes with increased (rather than decreased) expression in the human brain. PMID- 12547511 TI - Gene conversion drives GC content evolution in mammalian histones. AB - To examine the evolutionary influence of gene conversion on DNA base composition, I analysed an exhaustive dataset of histone paralogous genes from human and mouse. I show that those gene copies that belong to subfamilies of very similar sequences (presumably undergoing gene conversion) have a higher GC content than unique gene copies (presumably not undergoing gene conversion). Thus, it seems that gene conversion is a biased process that tends to increase the DNA GC content, a conclusion that has implications for the evolution of isochores in vertebrates. PMID- 12547512 TI - Origin of a substantial fraction of human regulatory sequences from transposable elements. AB - Transposable elements (TEs) are abundant in mammalian genomes and have potentially contributed to their hosts' evolution by providing novel regulatory or coding sequences. We surveyed different classes of regulatory region in the human genome to assess systematically the potential contribution of TEs to gene regulation. Almost 25% of the analyzed promoter regions contain TE-derived sequences, including many experimentally characterized cis-regulatory elements. Scaffold/matrix attachment regions (S/MARs) and locus control regions (LCRs) that are involved in the simultaneous regulation of multiple genes also contain numerous TE-derived sequences. Thus, TEs have probably contributed substantially to the evolution of both gene-specific and global patterns of human gene regulation. PMID- 12547513 TI - An evolutionary basis for scrapie disease: identification of a fish prion mRNA. AB - Infectious prion proteins cause neurodegenerative disease in mammals owing to the acquisition of an aberrant conformation. We cloned a Fugu rubripes gene that encodes a structurally conserved prion protein, and found rapid rates of molecular divergence among prions from different vertebrate classes, along with molecular stasis within each class. We propose that a directional trend in the evolution of prion sequence motifs associated with pathogenesis and infectivity could account for the origin of scrapie in mammals. PMID- 12547514 TI - Functional determinants of transcription factors in Escherichia coli: protein families and binding sites. AB - DNA-binding transcription factors regulate the expression of genes near to where they bind. These factors can be activators or repressors of transcription, or both. Thus, a fundamental question is what determines whether a transcription factor acts as an activator or a repressor? Previous research into this question found that a protein's regulatory function is determined by one or more of the following factors: protein-protein contacts, position of the DNA-binding domain in the protein primary sequence, altered DNA structure, and the position of its binding site on the DNA relative to the transcription start site. Although there are many aspects specific to different transcription factors, in this work we demonstrate that, in general, in the prokaryote Escherichia coli, a transcription factor's protein family is not indicative of its regulatory function, but the position of its binding site on the DNA is. PMID- 12547515 TI - Life cycles of successful genes. AB - By exploring time-series data from MEDLINE abstracts, we observe that only a few genes have been quoted with increasing frequency during the past 25 years. This is probably the result of selective pressure by the scientific community. Over the years, this selection has produced an extreme power law distribution of the information available for individual genes. Interestingly, those genes that are successfully selected are not necessarily the most important genes to the cell. To stress the implication of this finding we show that there is no correlation between a gene's impact in the scientific literature and its centrality in protein-interaction networks. PMID- 12547516 TI - The mouse Ink4a/Arf locus: a p53 pile-up at a tumour surveillance crossroads? PMID- 12547517 TI - Recombination or mutation rate heterogeneity? Implications for Mitochondrial Eve. AB - The study of mitochondrial DNA (mtDNA) has helped to demonstrate the African origin of our species and the relationship between living humans and the Neanderthals. mtDNA data have also been used to establish the time and route of major events in human history, such as the expansion of Neolithic farmers into Europe, and the settlement of the Pacific and the New World. However, it is becoming apparent that mtDNA evolution is more complex than previously believed. Anomalous mutation patterns perturb phylogenetic assumptions based on mtDNA data. Although they are frequently dismissed as sequencing errors or mutation hotspots, some of the anomalies have no satisfactory explanation. The mechanisms behind apparent mutation rate heterogeneity, or even possible mtDNA recombination, remain unknown. These issues need to be addressed, as they have profound consequences for the interpretation of mtDNA data. PMID- 12547518 TI - Patterning the fly eye: the role of apoptosis. AB - Early development in many tissues is characterized by a rapid expansion in cell number. Excess cells are removed through activation of their intrinsic apoptotic machinery. This over-expansion followed by selective removal is important for the sculpting of these tissues, and how specific cells are selected to die is one of the central questions in development. The Drosophila eye is a unique example of such patterning through cell death. Because of its remarkable reiterative design, the fly eye lends itself to studies of mutants with increased or decreased apoptosis. We know that the process of elimination of lattice cells is highly regulated. And we have learned that each ommatidial unit is involved in the life death decision of lattice cells through cell-cell signaling. But, we have yet to understand how this signaling is regulated spatially to result in such precision. In this article, we describe and speculate on the role of selective cell death during maturation of the fly eye. PMID- 12547519 TI - A polygenic basis for late-onset disease. AB - The biological basis of late-onset disease has been shaped by genetic factors subject to varying degrees of evolutionary constraint. Late-onset traits are not only more sensitive to environmental variation, owing to the breakdown of homeostatic mechanisms, but they also show higher levels of genetic variation than traits directly influencing reproductive fitness. The origin and nature of this variation suggests that current strategies are poorly suited to identifying genes involved in many complex diseases. PMID- 12547520 TI - Transient genetic asymmetry and cell fate in a bacterium. AB - Certain species of Gram-positive bacteria can initiate a developmental program that results in the formation of two daughter cells with different fates. One cell develops into a spore and the other cell undergoes programmed lysis, with each process being mediated by a cascade of cell-type-specific transcription factors. An early and critical step in this developmental pathway is the formation of a division septum near one pole, creating two compartments of different sizes. But how is this morphological asymmetry translated into the transcriptional asymmetry of the two compartments? Recent results suggest that the chromosomal position of the genes encoding several key components of the transcriptional regulatory network has an important role in this process. PMID- 12547521 TI - Yield and nutritive quality of sericea lespedeza (Lespedeza cuneata) and little bluestem (Schizachyrium scoparium) exposed to ground-level ozone. AB - Sericea lespedeza (Lespedeza cuneata cv. Interstate 76) and little bluestem (Schizachyrium scoparium cv. Aldous) were raised from seed in a glasshouse, transplanted into 5.7-l pots and placed into open-top chambers (OTC) on 6 June 1999. Following a 7-day adjustment period, each of six OTCs (duplicate OTCs per treatment) was ventilated with either air that had been carbon-filtered (CF) to remove ambient ozone (O3); non-filtered (NF), representative of ambient air; or enriched to twice-ambient O3 concentration (2X). Primary-growth forage was harvested on days 7, 32, 46, 59 and 72 following the start of fumigation, and regrowth forage from the first primary-growth harvest was harvested on days 36, 54 and 72 following the start of fumigation. Dry matter (DM) yield of either forage species did not differ among treatments except in the final regrowth period when yield of sericea lespedeza was greater for the NF than 2X O3 treatment. In vitro DM digestibility (IVDMD) and concentrations of crude protein (CP), soluble phenolics (SP) and condensed tannins (CT) in primary-growth sericea lespedeza did not differ between treatments, but NF primary-growth forage had higher concentration of protein-precipitating tannins (PPT) than did 2X primary growth forage. Concentrations of neutral detergent fiber (NDF), acid detergent fiber (ADF) and acid detergent lignin (ADL) were higher in NF and 2X than in CF primary-growth sericea lespedeza. Similarly, concentrations of NDF and ADL were higher, and IVDMD was lower for NF and 2X than for CF regrowth sericea lespedeza. Concentrations of ADF and ADL were lower, whereas IVDMD, in vitro NDF digestibility (IVNDFD) and concentrations of CP and SP were greater, in CF than in NF and 2X primary-growth little bluestem. Percentages IVDMD and IVNDFD and concentrations of CP and SP in NF primary-growth little bluestem were greater than those in forage exposed to 2X O3 treatment. No significant differences were observed among treatments in percentages IVDMD and IVNDFD, or concentrations of cell wall constituents or SP in little bluestem regrowth. Nutritive quality of little bluestem was decreased by < 2%, and that of sericea lespedeza by approximately 7% as a result of increased concentrations of cell wall constituents and decreased in vitro digestibility of NF and 2X compared with CF forages. Results indicate that existing and projected O3 levels can drive alterations in forage quality of select warm-season forages sufficient to have nutritional and economic implications for their utilization by ruminant herbivores. PMID- 12547522 TI - Solid phase speciation of arsenic by sequential extraction in standard reference materials and industrially contaminated soil samples. AB - Availability, mobility, (phyto)toxicity and potential risk of contaminants is strongly affected by the manner of appearance of elements, the so-called speciation. Operational fractionation methods like sequential extractions have been applied for a long time to determine the solid phase speciation of heavy metals since direct determination of specific chemical compounds can not always be easily achieved. The three-step sequential extraction scheme recommended by the BCR and two extraction schemes based on the phosphorus-like protocol proposed by Manful (1992, Occurrence and Ecochemical Behaviours of Arsenic in a Goldsmelter Impacted Area in Ghana, PhD dissertation, at the RUG) were applied to four standard reference materials (SRM) and to a batch of samples from industrially contaminated sites, heavily contaminated with arsenic and heavy metals. The SRM 2710 (Montana soil) was found to be the most useful reference material for metal (Mn, Cu, Zn, As, Cd and Pb) fractionation using the BCR sequential extraction procedure. Two sequential extraction schemes were developed and compared for arsenic with the aim to establish a better fractionation and recovery rate than the BCR-scheme for this element in the SRM samples. The major part of arsenic was released from the heavily contaminated samples after NaOH extraction. Inferior extraction variability and recovery in the heavily contaminated samples compared to SRMs could be mainly contributed to subsample heterogeneity. PMID- 12547524 TI - Persistent organochlorine contaminants in eggs of northern goshawk and Eurasian buzzard from northeastern Spain: temporal trends related to changes in the diet. AB - Persistent organochlorine compounds (pesticides and polychlorinated biphenyls) were determined in 24 northern goshawk (Accipiter gentilis) and eight Eurasian buzzard (Buteo buteo) samples of eggs collected between 1988 and 1999 in La Segarra (northeast Spain), in order to evaluate the changes in exposure and detrimental effects during this period. In the study area, both species exhibited similar levels of contamination, which may be related to their similar diet, mainly based on European rabbits (Oryctolagus cuniculus) in terms of biomass. The buzzard showed contamination levels similar to those found in other Spanish areas, but the levels found in the goshawk were much lower. The shell index in goshawk eggs was inversely correlated to concentration of p,p'-DDE. In late eighties, the concentrations of p,p'-DDE and heptachlor-epoxide in goshawk eggs were positively correlated to the biomass percentage of passeriforms in the diet. In goshawk samples, a decline in HCB concentration in the 1990s as compared to the 1980s was detected. Surprisingly, p,p'-DDE concentrations did not decline, as could be expected from the ban on DDT use. On the contrary, the highest p,p'-DDE concentrations were detected in some samples from the nineties, which also showed the lowest shell indices. This may be related to a severe reduction of rabbit population after 1989 that produced an increase in the consumption of passeriformes, which are known to accumulate higher levels of organochlorine compounds. Our study suggests that monitoring programs aiming to detect temporal trends in chemical contamination should take into account changes in diet composition before any conclusion can be drawn. PMID- 12547523 TI - A field method using microcosms to evaluate transfer of Cd, Cu, Ni, Pb and Zn from sewage sludge amended forest soils to Helix aspersa snails. AB - Juvenile Helix aspersa snails exposed in field microcosms were used to assess the transfer of Cd, Cu, Ni, Pb and Zn from forest soils amended with liquid and composted sewage sludge. Zn concentrations and contents were significantly higher in snails exposed to liquid and composted sludge after 5 and 7 weeks of exposure, when compared with control. Trends were less clear for the other metals. Present results show that Zn, among the cocktail of metallic trace elements (MTE) coming from sewage sludge disposal, represents the principal concern for food chain transfer and secondary poisoning risks. The microcosm design used in this experiment was well suited for relatively long-term (about 2 months) active biomonitoring with H. aspersa snails. The snails quickly indicated the variations of MTE concentrations in their immediate environment. Therefore, the present study provides a simple but efficient field tool to evaluate MTE bioavailability and transfer. PMID- 12547525 TI - Effects of elevated soil copper on phenology, growth and reproduction of five ruderal plant species. AB - The repeated use of copper (Cu) fungicides to control vine downy mildew, caused by Plasmopara viticola, has been responsible for the heavy increase of Cu concentration in the upper layers of vineyard soils. To determine the effects of elevated soil Cu on plant development, we created an artificial soil gradient with Cu enrichments ranging from 0 to 400 mg kg-1. On this gradient, and for five ruderal plant species commonly found in vineyards in southern France (Poa annua L., Dactylis glomerata L., Senecio vulgaris L., Hypochoeris radicata L., and Andryala integriflolia L.), we quantified survival, growth, and reproduction throughout one flowering season. High concentrations of Cu in the soil resulted in low survival, low total plant biomass, delay in flowering and fruiting, and low seed set. However, the effects differed among species. Furthermore, high soil Cu concentrations had contrasting effects on patterns of resource allocation depending on the plant species. PMID- 12547526 TI - Mussel transplantation and biomarkers as useful tools for assessing water quality in the NW Mediterranean. AB - Mussels, coming from an aquaculture farm located in a clean open bay, were transplanted to several stations of the bays of Nice and Cannes (NW Mediterranean) including a reference site for one month at three periods. Several biomarkers: activities of glutathione S-transferase (GST; exposure to organics), of catalase (exposure to oxidative stress) and of acetylcholinesterase (inhibited by some pesticides) and the lipid peroxidation (thiobarbituric acid reactive substances: TBARS) were measured in transplanted mussels. Cd, Cu and Zn concentrations were also measured as well as their condition index. The results demonstrated some seasonal variations in GST and catalase activities with higher levels in June compared to October. The condition index was also higher in June than in October. Principal component analyses performed with the whole set of data allowed to separate stations or groups of stations according to their responses. The mussels from the harbour of Nice were characterized by high TBARS levels and catalase activity in October 1999 whereas in the harbour of Cannes, animals presented very high copper concentrations and GST activities in June 2000. At the reference site, mussels generally presented low enzymatic activities (except AChE activity) and peroxidation levels and low heavy metal concentrations. PMID- 12547527 TI - N:P ratios, light limitation, and cyanobacterial dominance in a subtropical lake impacted by non-point source nutrient pollution. AB - A long-term (28-year) data set was used to investigate historical changes in concentrations of phosphorus (P), nitrogen (N), N:P ratios, and Secchi disk transparency in a shallow subtropical lake (Lake Okeechobee, Florida, USA). The aim was to evaluate changes in the risk of N2-fixing cyanobacterial blooms, which have infrequently occurred in the lake's pelagic zone. Predictions regarding bloom risk were based on previously published N:P ratio models. Temporal trends in the biomass of cyanobacteria were evaluated using phytoplankton data collected in 1974, 1989-1992, and 1997-2000. Concentrations of pelagic total P increased from near 50 microg l-1 in the mid-1970s to over 100 microg l-1 in the late 1990s. Coincidentally, the total N:P (mass) ratio decreased from 30:1 to below 15:1, and soluble N:P ratio decreased from 15:1 to near 6:1, in the lake water. Published empirical models predict that current conditions favor cyanobacteria. The observations confirm this prediction: cyanobacteria presently account for 50 80% of total phytoplankton biovolume. The historical decrease in TN:TP ratio in the lake can be attributed to a decreased TN:TP ratio in the inflow water and to a decline in the lake's assimilation of P, relative to N. Coincident with these declines in total and soluble N:P ratios, Secchi disk transparency declined from 0.6 m to near 0.3 m, possibly due to increased mineral turbidity in the lake water. Empirical models predict that under the turbid, low irradiance conditions that prevail in this lake, non-heterocystous cyanobacteria should dominate the phytoplankton. Our observations confirmed this prediction: non-N2-fixing taxa (primarily Oscillatoria and Lyngbya spp.) typically dominated the cyanobacteria community during the last decade. The only exception was a year with very low water levels, when heterocystous N2-fixing Anabaena became dominant. In the near shore regions of this shallow lake, low N:P ratios potentially favor blooms of N2 fixing cyanobacteria, but their occurrence in the pelagic zone is restricted by low irradiance and lack of stable stratification. PMID- 12547528 TI - Enhancement of dissolved phosphorus release from sediment to lake water by Microcystis blooms--an enclosure experiment in a hyper-eutrophic, subtropical Chinese lake. AB - To clarify the possible influence of Microcystis blooms on the exchange of phosphorus (P) between sediment and lake water, an enclosure experiment was conducted in the hypereutrophic subtropical Lake Donghu during July-September 2000. Eight enclosures were used: six received sediment while two were sediment free. In mid-August, Microcystis blooms developed in all the enclosures. There was a persistent coincidence between the occurrence of Microcystis blooms and the increase of both total P (TP) and soluble reactive P (SRP) concentrations in the water of the enclosures with sediments. In sediment-free enclosures, TP and SRP concentrations remained rather stable throughout the experiment, in spite of the appearance of Microcystis blooms. The results indicate that Microcystis blooms induced massive release of P from the sediment, perhaps mediated by high pH caused by intense algal photosynthesis, and/or depressed concentrations of nitrate nitrogen (NO3-N). PMID- 12547529 TI - The acanthocephalan Paratenuisentis ambiguus as a sensitive indicator of the precious metals Pt and Rh from automobile catalytic converters. AB - Recent studies revealed that intestinal acanthocephalans of fish can accumulate heavy metals to concentrations orders of magnitude higher than those in the host tissues or the aquatic environment. This significant heavy metal accumulation by acanthocephalans, even surpassing that of established free living accumulation bioindicators, encouraged us to study the bioavailability of the platinum-group metals (PGM) Pt and Rh for parasites. These precious metals are used in catalytic converters of cars for exhaust gas purification in Europe since the early 1980s. In addition to the beneficial effect in reducing the emission of CHx, CO and NOx of cars there is an increasing emission of these metals. However, it still remains unclear if these elements become accumulated in the biosphere and whether they affect the health of organisms. The present study reveals that in European eels (Anguilla anguilla) naturally infected with the eoacanthocephalan parasite Paratenuisentis ambiguus and experimentally exposed to ground catalytic converter material, the parasites take up and accumulate the catalytic active metals Pt and Rh whereas in the examined host tissues we found no metal uptake. Compared with the PGM concentrations in the water the worms contained 1600 times higher Rh and 50 times higher Pt concentrations. Thus, the parasites can be used as sentinel organisms reflecting even very low levels of precious metals. PMID- 12547530 TI - Assessing sediment toxicity and arsenite concentration with bacterial and traditional methods. AB - Three sediment samples LP (pool where logs are stored), LF (brook through landfill area), KN (Kaskesniemi) which is in Lake Pyhaselka downstream from the mill, were taken from an old sawmill area and one from the unpolluted Lake Hoytiainen. The arsenite concentration was measured by GFAAS and two arsenite biosensing bacterial strains Pseudomonas fluorescens OS8 (pTPT31) and Escherichia coli MC1061 (pTOO31). The toxicity of sediment and pore water samples was determined by using luminescent bacteria (Flash test) and, further, whole sediment toxicity was measured using 10 days growth test and 50 days emergency test with midges (Chironomus riparius). With the flash test a lowered EC50 value was found only in sediment LF (EC50=0.17 v/v%). The Flash test indicated that all sediment samples taken from the sawmill area were highly toxic to bacteria, whereas growth and the emergence of chironomids showed no effects in other samples than LF. The midges tolerate well the contaminated environment. In contrast, bioavailability of arsenite of sediment samples KN and LF was quite high determined using the biosensor-strains in a direct contact assay. The bioavailable fraction of sediment LP was 6-10% out of the total arsenite concentration obtained with GFAAS (0.46-0.77 microg g-1 dw). The results show that the choice of analysis method grossly affects the outcome without any of the method giving an incorrect result. Different methods measure different parameters of a toxic sample and can thus be used to complement each other. PMID- 12547531 TI - Environmentally acceptable effect of hydrogen peroxide on cave "lamp-flora", calcite speleothems and limestones. AB - Mosses, algae, and cyanobacteria (lamp-flora) colonize illuminated areas in show caves. This biota is commonly removed by a sodium hypochlorite solution. Because chlorine and other deleterious compounds are released into a cave environment during lamp-flora cleansing, hydrogen peroxide was tested as an alternative agent. In a multidisciplinary study conducted in the Kateinska Cave (Moravian Karst, Czech Republic), 12 algae- and cyanobacteria taxons and 19 moss taxons were detected. The threshold hydrogen peroxide concentration for the destruction of this lamp-flora was found to be 15 vol.%. Based on laboratory experiments in stirred batch reactors, the dissolution rates of limestones and calcite speleothems in water were determined as 3.77 x 10-3 and 1.81 x 10-3 mol m-2 h-1, respectively. In the 15% peroxide solution, the limestone and speleothem dissolution rates were one order of magnitude higher, 2.00 x 10-2 and 2.21 x 10-2 mol m-2 h-1, respectively. So, the peroxide solution was recognised to attack carbonates somewhat more aggressively than karst water. In order to prevent the potential corrosion of limestone and speleothems, the reaching of preliminary peroxide saturation with respect to calcite is recommended, for example, by adding of few limestone fragments into the solution at least 10 h prior to its application. PMID- 12547532 TI - Organochlorine contaminant and stable isotope profiles in Arctic fox (Alopex lagopus) from the Alaskan and Canadian Arctic. AB - Arctic fox (Alopex lagopus) is a circumpolar species distributed across northern Canada and Alaska. Arctic fox muscle and liver were collected at Barrow, AK, USA (n=18), Holman, NT, Canada (n=20), and Arviat, NU, Canada (n=20) to elucidate the feeding ecology of this species and relate these findings to body residue patterns of organochlorine contaminants (OCs). Stable carbon (delta 13C) and nitrogen (delta 15N) isotope analyses of Arctic fox muscle indicated that trophic position (estimated by delta 15N) is positively correlated with increasing delta 13C values, suggesting that Arctic fox with a predominantly marine-based foraging strategy occupy a higher trophic level than individuals mostly feeding from a terrestrial-based carbon source. At all sites, the rank order for OC groups in muscle was polychlorinated biphenyls (Sigma PCB) > chlordane-related compounds (Sigma CHLOR) > hexachlorocyclohexane (Sigma HCH) > total toxaphene (TOX) > or = chlorobenzenes (Sigma ClBz) > DDT-related isomers (Sigma DDT). In liver, Sigma CHLOR was the most abundant OC group, followed by Sigma PCB > TOX > Sigma HCH > Sigma ClBz > Sigma DDT. The most abundant OC analytes detected from Arctic fox muscle and liver were oxychlordane, PCB-153, and PCB-180. The comparison of delta 15N with OC concentrations indicated that relative trophic position might not accurately predict OC bioaccumulation in Arctic fox. The bioaccumulation pattern of OCs in the Arctic fox is similar to the polar bear. While Sigma PCB concentrations were highly variable, concentrations in the Arctic fox were generally below those associated with the toxicological endpoints for adverse effects on mammalian reproduction. Further research is required to properly elucidate the potential health impacts to this species from exposure to OCs. PMID- 12547533 TI - Removal of pollutants from surface water and groundwater by nanofiltration: overview of possible applications in the drinking water industry. AB - During the last decade, nanofiltration (NF) made a breakthrough in drinking water production for the removal of pollutants. The combination of new standards for drinking water quality and the steady improvement of the nanofiltration process have led to new insights, possible applications and new projects on lab-scale, pilot scale and industrial scale. This paper offers an overview of the applications in the drinking water industry that have already been realised or that are suggested on the basis of lab-scale research. Applications can be found in the treatment of surface water as well as groundwater. The possibility of using NF for the removal of hardness, natural organic material (NOM), micropollutants such as pesticides and VOCs, viruses and bacteria, salinity, nitrates, and arsenic will be discussed. Some of these applications have proven to be reliable and can be considered as known techniques; other applications are still studied on laboratory scale. Modelling is difficult due to effects of fouling and interaction between different components. The current insight in the separation mechanisms will be briefly discussed. PMID- 12547534 TI - Changes of copper speciation in maize rhizosphere soil. AB - Chemical forms of copper in the rhizosphere and bulk soil of maize were investigated using rhizobox cultivation and sequential extraction techniques. The copper accumulations were also determined. The results demonstrated that there were continuous changes in copper fractionation within the maize rhizosphere. Initially, the amount of exchangeable copper increased before dropping below the initial level after 40 days or so. Carbonate associated copper followed a similar trend of change, but with a slower pace than the exchangeable copper. The increase in carbonate associated copper only become evident after 30 days, with the net loss occurring after 60 days. There were also initial increases in oxide bound copper as well as decreases in the organic matter associated copper, both followed by a turnover after 40-50 days. The accumulation of copper in the maize plant was found to be biomass dependent. The amount of accumulated copper absorbed in the plant material exceeded the initial quantity of the exchangeable copper in the soil, revealing a transformation from less bioavailable to more bioavailable fractions. During cultivation, decreases in redox potential and increases in pH, dissolved organic carbon (DOC), and microbial activity in the maize rhizosphere were observed. The change in copper speciation may result from root-induced changes in DOC, redox potential, and microbial activity in the rhizosphere. PMID- 12547535 TI - Transparency at IARC. PMID- 12547536 TI - Screening interval for retinopathy in type 2 diabetes. PMID- 12547537 TI - A major new test for plague. PMID- 12547538 TI - New insights into EBV-associated post-transplant lymphoproliferative disease. PMID- 12547539 TI - The ins and outs of fetal DNA in maternal plasma. PMID- 12547540 TI - WHO's next Director-General: time to decide. PMID- 12547541 TI - Incidence of sight-threatening retinopathy in patients with type 2 diabetes in the Liverpool Diabetic Eye Study: a cohort study. AB - BACKGROUND: Incidence data on which to base targets and protocols for screening for sight-threatening diabetic retinopathy are few. We aimed to investigate yearly and cumulative incidence of any retinopathy, maculopathy, and sight threatening diabetic retinopathy in patients with type 2 diabetes in an established systematic programme and to calculate optimum screening intervals according to retinopathy grade at baseline. METHODS: We investigated all patients with type 2 diabetes registered with enrolled general practices (except those who were attending an ophthalmologist) who had retinopathy data available at baseline and at least one further screening event. To screen patients, we used non stereoscopic three-field mydriatic photography and modified Wisconsin grading. Sight-threatening diabetic retinopathy was defined as moderate preproliferative retinopathy or worse, or clinically significant maculopathy in either or both eyes. FINDINGS: Results were obtained from 20 570 screening events. Yearly incidence of sight-threatening diabetic retinopathy in patients without retinopathy at baseline was 0.3% (95% CI 0.1-0.5) in the first year, rising to 1.8% (1.2-2.5) in the fifth year; cumulative incidence at 5 years was 3.9% (2.8 5.0). Rates of progression to sight-threatening diabetic retinopathy in year 1 by baseline status were: background 5.0% (3.5-6.5), and mild preproliferative 15% (10.2-19.8). For a 95% probability of remaining free of sight-threatening diabetic retinopathy, mean screening intervals by baseline status were: no retinopathy 5.4 years (95% CI 4.7-6.3), background 1.0 years (0.7-1.3), and mild preproliferative 0.3 years (0.2-0.5). INTERPRETATION: A 3-year screening interval could be safely adopted for patients with no retinopathy, but yearly or more frequent screening is needed for patients with higher grades of retinopathy. PMID- 12547542 TI - Effect of recombinant activated factor VII on perioperative blood loss in patients undergoing retropubic prostatectomy: a double-blind placebo-controlled randomised trial. AB - BACKGROUND: Recombinant activated factor VII (factor VIIa) has prohaemostatic effects in bleeding patients with coagulation abnormalities. We aimed to test the hypothesis that recombinant factor VIIa could reduce perioperative blood loss in patients with normal coagulation systems. Therefore, we assessed safety and efficacy of this drug in patients undergoing retropubic prostatectomy, which is often associated with major blood loss and need for transfusion. METHODS: In a double-blind, randomised placebo-controlled trial, we recorded blood loss and transfusion requirements in 36 patients undergoing retropubic prostatectomy, who were randomised to receive an intravenous bolus of recombinant factor VIIa (20 microg/kg or 40 microg/kg) or placebo in the early operative phase. FINDINGS: Median perioperative blood loss was 1235 mL (IQR 1025-1407) and 1089 mL (928 1320) in groups given recombinant factor VIIa 20 microg/kg and 40 microg/kg, respectively, compared with 2688 mL (1707-3565) in the placebo group (p=0.001). Seven of twelve placebo-treated patients were transfused, whereas no patients who received 40 microg/kg recombinant factor VIIa needed transfusion. The odds ratio for receiving any blood product in patients treated with recombinant factor VIIa compared with control patients was 0 (95% CI 0.00-0.33) No adverse events arose. INTERPRETATION: An injection of recombinant factor VIIa can reduce perioperative blood loss and eliminate the need for transfusion in patients undergoing major surgery. PMID- 12547543 TI - Circulating endothelial cells as markers for ANCA-associated small-vessel vasculitis. AB - BACKGROUND: Histological findings in small-vessel vasculitis associated with antineutrophil cytoplasmic antibodies (ANCAs) suggest that damaged endothelial cells undergo necrosis and detachment from the basement membrane. We postulated that isolation of these cells from peripheral blood might provide a novel marker of the disease and elucidate pathogenetic events. METHODS: 18 patients with active ANCA-associated vasculitis, 20 patients in remission, 20 healthy controls, 12 patients with infection, and 12 patients with glomerular disease not associated with ANCA were studied. Endothelial cells were isolated from peripheral blood by use of Dynabeads coated with antibodies against CD146, and were stained for von Willebrand factor (vWF), CD31, and Ulex Europaeus lectin 1 (UEA-1). Tissue-factor immunocytochemistry and assays for markers of apoptosis and necrosis were also done. FINDINGS: Few circulating endothelial cells were seen in healthy controls (0-20 cells/mL, median 5 cells/mL), patients with infection (0-16 cells/mL, median 8 cells/mL), and patients with non-ANCA glomerulonephritis (0-21 cells/mL, median 4 cells/mL). By contrast, large numbers of circulating endothelial cells were detected in patients with active vasculitis (20-5700 cells/mL, median 136 cells/mL, p<0.0001 when compared with healthy controls). Cell numbers fell substantially during 6 months of successful immunosuppressive treatment among those with active disease. Patients in remission had moderately raised cell numbers (0-60 cells/mL, median 16 cells/mL). 84% of cells obtained from patients with active disease stained positive for annexin/propidium iodide and 86% stained tissue factor positive, indicating a necrotic and procoagulant phenotype. INTERPRETATION: Circulating endothelial cells are a novel marker of active ANCA-associated small-vessel vasculitis. The clinical use of this tool and the pathogenic mechanisms leading to these findings require further investigation. PMID- 12547544 TI - Development and testing of a rapid diagnostic test for bubonic and pneumonic plague. AB - BACKGROUND: Plague is often fatal without prompt and appropriate treatment. It affects mainly poor and remote populations. Late diagnosis is one of the major causes of human death and spread of the disease, since it limits the effectiveness of control measures. We aimed to develop and assess a rapid diagnostic test (RDT) for plague. METHODS: We developed a test that used monoclonal antibodies to the F1 antigen of Yersinia pestis. Sensitivity and specificity were assessed with a range of bacterial cultures and clinical samples, and compared with findings from available ELISA and bacteriological tests for plague. Samples from patients thought to have plague were tested with the RDT in the laboratory and by health workers in 26 pilot sites in Madagascar. FINDINGS: The RDT detected concentrations of F1 antigen as low as 0.5 ng/mL in up to 15 min, and had a shelf life of 21 days at 60 degrees C. Its sensitivity and specificity were both 100%. RDT detected 41.6% and 31% more positive clinical specimens than did bacteriological methods and ELISA, respectively. The agreement rate between tests done at remote centres and in the laboratory was 89.8%. With the combination of bacteriological methods and F1 ELISA as reference standard, the positive and negative predictive values of the RDT were 90.6% and 86.7%, respectively. INTERPRETATION: Our RDT is a specific, sensitive, and reliable test that can easily be done by health workers at the patient's bedside, for the rapid diagnosis of pneumonic and bubonic plague. This test will be of key importance for the control of plague in endemic countries. PMID- 12547545 TI - Target cells of Epstein-Barr-virus (EBV)-positive post-transplant lymphoproliferative disease: similarities to EBV-positive Hodgkin's lymphoma. AB - BACKGROUND: Epstein-Barr virus (EBV)-associated post-transplant lymphoproliferative disease (PTLD) encompasses a histologically broad range of lesions, arising from the expanded pool of EBV-infected B cells in the immunocompromised host. Identification of the precise cellular origin of these tumours could clarify their pathogenesis. METHODS: Of 13 cases of EBV-positive cases of PTLD characterised by histological analysis, pattern of EBV gene expression, and clinical course, 11 had monoclonal or biclonal lesions in which we determined the progenitor B cell by immunoglobulin heavy chain (IgH) genotyping. RESULTS: Two tumours had a naive B cell genotype and two showed patterns of IgH somatic mutation typical of antigen-selected (post-germinal centre) memory cells. All four arose early post-transplant and expressed the markers of EBV transformation--Epstein-Barr nuclear antigen (EBNA) 2 and latent membrane protein (LMP) 1. However, seven tumours, either of early or late onset and including some with downregulated EBNA 2 and LMP 1, arose from post-germinal cells with randomly mutated or sterile IgH genotypes usually incompatible with B cell survival in vivo. INTERPRETATION: PTLD can arise from a broad range of target B cells and not only from the pool of antigen-selected memory cells that EBV generally colonises in immunocompetent individuals. Tumour development seems frequently associated with the EBV-induced rescue and expansion of B cells that have failed the physiological process of germinal centre selection into memory. This finding shows an unexpected connection between pathogenesis of PTLD and that of EBV-positive Hodgkin's lymphoma, another B-cell malignancy of atypical post germinal-centre cell origin. PMID- 12547546 TI - A loose sternotomy wire. PMID- 12547547 TI - Anticonvulsants for Creutzfeldt-Jakob disease? PMID- 12547548 TI - Low back pain in rural Tibet. AB - In a baseline assessment of 30 rural villages surrounding Shigatse City, Tibet, many people, especially women, identified low back pain as a serious health problem. Consequently, we aimed to establish the prevalence of such pain and to develop appropriate interventions. We did a cross-sectional study of the prevalence of low back pain and related functional disability using two-stage random cluster sampling. We included 499 adults aged at least 15 years from 19 villages. The point prevalence of low back pain was 34.1% (95% CI 27.9-40.3% [170 people]); the 12-month prevalence was 41.9% (35.5-48.3% [209 people]). 100 (20%) villagers had substantial functional disability associated with low back pain. Low back pain is likely to be an important and under recognised problem in rural societies like Tibet. PMID- 12547549 TI - Adiponectin and protection against type 2 diabetes mellitus. AB - Adiponectin is an adipocyte-derived peptide, which has anti-inflammatory and insulin-sensitising properties. We designed a nested case-control study to assess whether baseline adiponectin concentrations in plasma are independently associated with risk of type 2 diabetes. We found that adiponectin concentrations in plasma were lower among individuals who later developed type 2 diabetes than among controls (mean 5.34 microg/mL [SD 3.49] vs 6.87 microg/mL [4.58], p<0.0001). High concentrations of adiponectin were associated with a substantially reduced relative risk of type 2 diabetes after adjustment for age, sex, waist-to-hip ratio, body-mass index, smoking, exercise, alcohol consumption, education, and glycosylated haemoglobin A(1c) (odds ratio 4th vs 1st quartile 0.3 [95% CI 0.2-0.7], p=0.0051). We conclude that adiponectin is independently associated with a reduced risk of type 2 diabetes in apparently healthy individuals. PMID- 12547550 TI - A tumour that secretes glucagon-like peptide-1 and somatostatin in a patient with reactive hypoglycaemia and diabetes. AB - Glucagon-like peptide 1 (GLP-1), an insulinotropic hormone normally synthesised in the intestinal mucosa and released in response to a meal, is essential for normal glucose homoeostasis. There is much interest in the use of GLP-1 to treat diabetes, since the risk of hypoglycaemia is thought to be low. We report an instance of a 45-year-old woman with a GLP-1 and somatostatin secreting neuroendocrine tumour who presented with reactive hypoglycaemia and hyperglycaemia, but who was subsequently cured by surgery. This case, of a neuroendocrine tumour secreting GLP-1 and causing reactive hypoglycaemia, indicates a potential adverse effect of GLP-1 therapy for diabetes. PMID- 12547551 TI - Whipple's disease. AB - Whipple's disease, or intestinal lipodystrophy, is a systemic infectious disorder affecting mostly middle-aged white men. Patients present with weight loss, arthralgia, diarrhoea, and abdominal pain. The disease is commonly diagnosed by small-bowel biopsy; the appearance of the sample is characterised by inclusions in the lamina propria staining with periodic-acid-Schiff, which represent the causative bacteria. Tropheryma whipplei has been classified as an actinomycete and has been propagated in vitro, which allows the possibility of improving diagnostic strategies, for example through antibody-based detection of the bacillus on duodenal tissue or in circulating monocytes. Cell-mediated immunity in active and inactive Whipple's disease has subtle defects that might predispose some individuals to symptomatic infection with this bacillus, which probably occurs ubiquitously. Although most patients respond well to empirical antibiotic treatment, some with relapsing disease have a poor outlook. The recent findings and concerted research might allow development of new strategies for diagnosis, treatment, and monitoring of patients with Whipple's disease. PMID- 12547552 TI - Drug-eluting stents in vascular intervention. AB - CONTEXT: Restenosis is the most important long-term limitation of stent implantation for coronary artery disease, occurring in 15-60% of patients. In stent restenosis, a refractory coronary lesion resulting from neointimal hyperplasia, challenges both vascular biologist and interventional cardiologist. Various drugs and devices have been used tried to overcome restenosis but are not particularly successful. Over 1500000 percutaneous coronary interventions are done annually. Restenosis is not only important clinically but also for its impact on health-care costs. STARTING POINT: Growth and migration of vascular smooth-muscle cells result in neointimal proliferation after vascular injury and are the key mechanism of in-stent restenosis. The rationale of the most recent approaches to restenosis (eg, brachytherapy and immunosuppressive agents) arises from the similarity between tumour-cell growth and the benign tissue proliferation which characterises intimal hyperplasia. Several immunosuppressants have been tested for their potential to inhibit restenosis, with the novel strategy of administering the drug via a coated stent platform. Local drug delivery achieves higher tissue concentrations of drug without systemic effects, at a precise site and time. The first multicentre trial with stents coated with sirolimus was by Marie-Claude Morice and colleagues (N Engl J Med 2002; 346: 1773 80). In a trial of 238 patients, restenosis of 50% or more at 6 months was 0% and 27% with sirolimus or normal stents (p<0.001), respectively, after percutaneous revascularisation. Muzaffer Degertekin and colleagues (Circulation 2002; 106: 1610-13) present data on 2-year follow-up of 15 patients who had been implanted with the sirolimus stent in another study, and confirm persistent inhibition of restenosis and an absence of unexpected adverse events. WHERE NEXT? Local application of antiproliferative agents is a promising technique and research is developing. Other agents with potential benefits (eg, statins, local gene therapy, adenovirus-mediated arterial gene-transfer, L-arginine, abciximab, angiopeptin, recombinant pegylated hirudin, and hiloprost) as well as improvements in polymer technology (biodegradable smart polymers, coatings for multiple-drug release) are under evaluation. The clinical impact of the elimination of restenosis may influence the approach to coronary artery disease, the future of cardiac surgery, and health-care economics in cardiology. PMID- 12547553 TI - Ethical reappraisal of 15 years of cord-blood transplantation. AB - Since the first successful use of cord blood as source of haemopoietic stem cells for transplantation in 1988, more than 2000 patients with malignant or non malignant disorders have been treated with this procedure. Collection and storage of cord blood has prompted ethical considerations, mainly dealing with the issues of autonomy in making decisions about donation of cord blood, and of privacy and confidentiality in the tests required before use of placental cells for transplantation. The ethical implications of possible storage of cord-blood cells for autologous use has also been discussed. Preimplantation selection of HLA matched embryos to obtain a donor of cells for cord-blood transplantation of a sibling with a life-threatening disease has raised the issue of the extent to which this approach complies with the principles of bioethics. PMID- 12547554 TI - Conjugal Parkinson's disease. PMID- 12547555 TI - Open letter to WHO Executive Board and candidates for Director General. PMID- 12547556 TI - Long-term effects of hormone replacement therapy. PMID- 12547557 TI - Long term-effects of hormone replacement therapy. PMID- 12547559 TI - Long term-effects of hormone replacement therapy. PMID- 12547560 TI - Assessing biomedical science. PMID- 12547561 TI - "Will not" or "does"? PMID- 12547562 TI - Angiogenesis with recombinant fibroblast growth factor-2 for claudication. PMID- 12547564 TI - Diet and risk of cancer. PMID- 12547566 TI - Diet and risk of cancer. PMID- 12547567 TI - Diet and risk of cancer. PMID- 12547568 TI - Defining physical inactivity. PMID- 12547569 TI - Minimum pesticide list for the developing world. PMID- 12547570 TI - Minimum pesticide list for the developing world. PMID- 12547571 TI - Herpes encephalitis. PMID- 12547572 TI - Emotional problems of Palestinian children living in a war zone. PMID- 12547573 TI - Ocular tuberculosis. PMID- 12547574 TI - Avoidance of bioflavonoid supplements during pregnancy. PMID- 12547576 TI - Troubling trial design. PMID- 12547577 TI - Prediction of long-term survival rates of cancer patients. PMID- 12547578 TI - Sildenafil for lung fibrosis and pulmonary hypertension. PMID- 12547580 TI - 5-year follow-up of replantation of penis and testis in a child. PMID- 12547581 TI - Medical education activities of the Association of American Medical Colleges. PMID- 12547582 TI - Colour neglect in an artist. PMID- 12547583 TI - Adult learning and CME. PMID- 12547589 TI - Mechanisms of vaccine adjuvant activity: initiation and regulation of immune responses by vaccine adjuvants. PMID- 12547590 TI - Recombinant congenic strains of mice: a new tool for the genetic dissection of efficacy and toxicity of adjuvants. AB - The efficacy of immunization of a vaccine depends on an antigen, an adjuvant, and on the expression of multiple genes in the host. The responsiveness of various strains of mice to adjuvants is therefore dependent on their genetic background present as a complex, multigenic trait similarly as in man. We have recently developed a gene-discovery platform, termed recombinant congenic strains (RCS), that greatly facilitates the dissection, localization and characterization of genes that mediate complex traits such as responsiveness to adjuvants. These recombinant congenic mice, which were constructed from two progenitor strains (A/J and C57BL/6) that are phenotypically different for several spontaneous or infectious diseases, were generated such that they carry 13.5% of the one genome in 85% of the other genome. The use of these RCS mice therefore enables a more efficient identification of genes that mediate the responsiveness of the adjuvant. PMID- 12547591 TI - The adjuvant OM-174 induces both the migration and maturation of murine dendritic cells in vivo. AB - The aim of this study was to test the capacity of the novel adjuvant OM-174, a lipid A analog, to induce the migration and the maturation of murine dendritic cells (DC) in vivo, a step which is considered as the initiation of the adaptive immune response. BALB/c mice were injected intravenously or subcutaneously with OM-174. The spleen and popliteal lymph nodes were harvested, and analyzed for DC localization and phenotype. The data presented here clearly show that, OM-174 induces the migration of DC from the periphery to the T cell areas of lymphoid organs, and their maturation into cells expressing high levels of MHC class II and co-stimulatory molecules, with a potency close to that of Escherichia coli lipopolysaccharide (LPS). PMID- 12547592 TI - Low effectiveness of DNA vaccination against HER-2/neu in ageing. AB - We evaluated the effectiveness of vaccination with a HER-2/neu DNA plasmid to induce protective immunity against HER-2/neu overexpressing syngeneic TUBO tumour cells in old ages. Young and old Balb/c mice received three immunizations with a pCMVneuNT DNA plasmid and, successively, were challenged with TUBO cells. Young mice were completely protected whereas less than 60% protection was observed in old mice. Anti-p185(neu) antibodies were found in the sera from both young and old immunized mice, even if antibody production was significantly higher in young in comparison with old mice. Similarly, higher anti-p185(neu) lymphocyte proliferation was induced in young than in old mice. No anti-p185(neu) cytotoxicity was found in lymphocytes from old animals. We conclude that anticancer DNA vaccination has a lower effectiveness in old than in young ages. PMID- 12547593 TI - Analysis of the role of vaccine adjuvants in modulating dendritic cell activation and antigen presentation in vitro. AB - We have studied the effects of adjuvant formulations on the activation and antigen-presenting functions of bone marrow-derived dendritic cells (DCs). While LPS could induce high-level expression of MHC Class II and co-stimulator molecules on DCs, it did not enhance antigen presentation to co-stimulation independent DO11.GFP T hybridoma cells. In contrast, alum, NISV and PLGA formulations failed to activate DCs, but NISV and PLGA could enhance antigen presentation efficiency by 10-100-fold. Irrespective of the previously described antigen release characteristics of each adjuvant, antigen presentation peaked at 6h and waned thereafter for all formulations. Given the importance of DCs in the activation of nai;ve T cell responses, these studies suggest that as yet undefined pathways of DC activation in vivo may underlie the activity of alum, PLGA and NISV adjuvants. Furthermore, as NISV and PLGA do not appear to act as slow-release systems in DCs, the ability of these particulate systems to induce high levels of antigen presentation by DCs probably has a more significant role in their adjuvant activity. PMID- 12547594 TI - Maturation of human dendritic cells induced by the adjuvant cholera toxin: role of cAMP on chemokine receptor expression. AB - Cholera toxin (CT) is a very effective adjuvant for mucosal vaccination. It binds to cells through its B subunit and induces intracellular increase of cAMP through the A subunit. We previously showed that CT induces maturation of human dendritic cells (DCs) and this may account for its adjuvant property. Here, we investigated the role of the A subunit on DCs maturation by using forskolin, a cAMP inducer. The results show that although cAMP does not stimulate full maturation of DCs it induces upregulation of the chemokine receptors CXCR4 and CCR7. PMID- 12547595 TI - MHC class II engagement by its ligand LAG-3 (CD223) leads to a distinct pattern of chemokine and chemokine receptor expression by human dendritic cells. AB - Upon stimulation by infectious agent products, dendritic cells (DC) become activated, express high levels of class I and class II antigens, CD80, CD86 and CD83 and migrate to secondary lymphoid organs where they can prime naive CD4 helper and CD8-cytotoxic T-cells. Cognate CD4(+) T-cell help mediated by CD40L along with DC stimulation with another T-cell effector molecule, termed lymphocyte activated gene-3 (LAG-3 or CD223, a ligand for MHC class II) have been shown to induce this maturation process. Both CD40L and LAG-3 have been used as vaccine adjuvants to induce CTL and CD4 Th1 responses. Here, we studied the effect of a soluble LAG-3Ig molecule on the chemokine and chemokine receptor profile of human immature monocyte-derived DC. LAG-3Ig, unlike CD40L, induced an inflammatory signal in terms of IL-8 and MIP-1alpha/CCL3 production and, in contrast to LPS, induced production of chemokines (MDC/CCL22 and TARC/CCL17) known to direct the migration of maturing DC to lymph nodes. In LAG-3-matured DC, surface expression of CCR5 (a receptor for MIP-1alpha/CCL3) was down-regulated and CCR7 (a receptor for MIP-3beta and SLC) was up-regulated. However, LAG-3 matured, but not LPS- or CD40L-matured DC retained their capacity to migrate in chemotaxis chambers and to respond to MIP-1alpha. Altogether, these data represent the first evidence that MHC class II signaling may affect DC migration to secondary lymphoid tissues. PMID- 12547596 TI - Dendritic cell KLH loading requirements for efficient CD4+ T-cell priming and help to peptide-specific cytotoxic T-cell response, in view of potential use in cancer vaccines. AB - This study focuses on a Keyhole Limpet Hemocyanin (KLH) adjuvant strategy to augment efficacy of dendritic cell-based vaccines that use class I-restricted peptides. Requirements for loading dendritic cells (DC) with KLH were first determined in order to optimally prime CD4(+) T cells. These KLH-loaded cells were pulsed with antigenic peptide and cultured with T cells to induce a cytotoxic T lymphocyte (CTL) response against the peptide. Such a concomitant presentation of KLH and peptide by the same DC strongly augmented the peptide specific CTL response, as compared to the response induced by DC pulsed with the peptide alone. This adjuvant effect was more pronounced for poorly immunogenic antigens. The use of optimised peptide and KLH-loaded DC may improve the efficacy of therapeutic anti-tumour peptide vaccination. PMID- 12547597 TI - Generation of antigen-loaded dendritic cells in a serum-free medium using different cytokine combinations. AB - Dendritic cells (DCs) are antigen-presenting cells that play a critical role in the induction of cytotoxic T-lymphocytes. An optimal method for the generation of DC for clinical use remains to be established. The aim of our study was to find an optimal cytokine combination for DC generation from peripheral blood stem cells (PBSC) and peripheral blood mononuclear cells (PBMC) in serum-free conditions. Serial immunophenotyping enabled us to observe changes in DC content during the culture as well as the development of maturation and activation markers. As a source for DC culture, we used PBSC from patients with multiple myeloma after stem cell mobilization using cyclophosphamide and G-CSF, or PBMC from healthy donors without mobilization. The cells were cultured in a serum-free medium with different cytokine combinations including GM-CSF, TNF-alpha, Flt-3, CD40L, IFN-gamma, IL-1alpha, IL-6, PGE1, and IL-4. The cell cultures were evaluated by immunophenotyping. For PBMC, interleukin-12 assay was performed. For PBSC, the yield of DC as determined by CD83+ cell count ranged from 0. 6 x 10(5) to 30.1 x 10(4) (mean: 9.4 x 10(4)) of DC generated per 1 x 10(6) of initially plated nucleated cells from apheresis. This yield corresponded to (0.3-19.1) x 10(5) (mean: 4.3 x 10(5)) per 1 x 10(6) of CD34+ cells in the apheresis products. For PBMC, the yield was (0.4-24.8) x 10(4) (mean: 2.4 x 10(4)) of DC generated per 1 x 10(6) of initially plated mononuclear cells from venous blood. The cultured cells expressed the mature immunophenotype. No significant differences in cell yield or immunophenotype were detected when comparing different cytokine combinations. PMID- 12547598 TI - Liposomal delivery of antigen to human dendritic cells. AB - This study investigated whether formulation of antigen in mannosylated liposomes enhanced uptake and activation of dendritic cells (DC) and increased the ability of DC to induce primed T cell proliferation compared to formulation of antigen in unmodified liposomes or in solution. Immature human DC were generated from peripheral blood monocytes cultured with GM-CSF and IL-4. Uptake of antigen by DC and the degree of expression of the cell surface markers MHC class II, CD80, CD86 and the DC maturation marker CD83, was investigated by flow cytometry following incubation with liposomes or solution containing FITC-conjugated antigen. Exposure to liposomes containing FITC-ovalbumin resulted in enhanced expression of cell surface markers when compared to exposure to antigen in solution. Expression was highest following exposure to mannosylated liposomes. Mannosylated liposomes containing tetanus toxoid (TT) stimulated primed T cell proliferation more effectively than TT-neutral liposomes or TT-solution. This work suggests that mannosylated liposomes provide a versatile delivery vehicle for initiating enhanced immune responses to encapsulated peptide or protein vaccines. PMID- 12547599 TI - Interleukin-2 and dendritic cells as adjuvants for surgical therapy of tumours associated with human papillomavirus type 16. AB - Moderately immunogenic HPV 16-associated tumours TC-1 (MHC class I(+), HPV 16 E6/E7(+), G12V Ha-ras(+)) and MK16/1/III ABC (MHC class I(-), HPV 16 E6/E7(+), G12V Ha-ras(+)), both of the H-2(b) haplotype and transplanted in syngeneic mice, were used to examine the adjuvant effects of IL-2 and dendritic cells for surgical therapy. Mice were inoculated s.c. with the respective tumour cells, and when the tumours reached 8-12 mm in diameter, they were extirpated. Three days after surgery, the experimental mice were treated with IL-2, IL-2 gene-modified tumour vaccines, or dendritic cells, injected s.c. to the site of previous surgery. It has been found in both, MHC class I(+) and MHC class I(-) tumours that the recombinant IL-2 and IL-2 gene-modified vaccines substantially reduced the tumour recurrence rate and inhibited growth of tumour recurrences. The dendritic cells were significantly effective only in mice with surgical minimal residual TC-1 (MHC class I(+)) tumour disease and when injected before they have reached the terminal stage of their differentiation. PMID- 12547600 TI - Immunopotentiating heat shock proteins: negotiators between innate danger and control of autoimmunity. AB - Heat shock proteins (hsps) are known to be immunodominant antigens of bacteria. Hsps are evolutionarily strongly conserved proteins present in all eukaryotic and prokaryotic cellular organisms and upregulated by several forms of stress. Despite (the paradigm of) self-tolerance, hsp-epitopes homologous to endogenous host hsp sequences have been implicated as T cell epitopes to endow crossreactive, hsp-specific T cells with the capacity to regulate inflammation, such as in experimentally induced autoimmune diseases. Such T cells were found to produce regulatory cytokines like IL10, in contrast to T cells induced with other conserved microbial proteins that are not upregulated by stress. Hsps have been implicated in immune regulation not only as upregulated targets of adaptive immunity during inflammatory stress, but recently also as triggering factors for innate immunity through activation via Toll-like receptors (TLRs). PMID- 12547601 TI - Induction of heterosubtypic immunity to influenza A virus using a DNA vaccine expressing hemagglutinin-C3d fusion proteins. AB - Cross-protection between different subtypes of influenza A virus has been attributed to heterosubtypic immunity (HSI). Although, HSI can occur in the absence of anti-HA or anti-NA antibodies, HSI seems to be mediated, in part, by cross-reactive antibodies. In this study, we examined the effects of a DNA vaccine expressing an influenza HA fused to three copies of murine C3d of complement (HA-mC3d(3)). HA-mC3d(3) elicited heterosubtypic immunity more efficiently than non-fused forms of HA and protected mice from lethal challenge of influenza with different subtypes. Plasmid encoding for various forms of HA were constructed from two influenza strains, A/Puerto Rico/8/34 (H1N1) or A/Aichi/2/68-x31 (H3N2). Vaccinated mice were analyzed for enhancement of anti-HA titers, affinity maturation of antibody, hemagglutinin-inhibition activity, and altered cytokine profiles. The HA-mC3d(3)-DNA vaccinated mice were protected from heterologous influenza challenge, even though sera from these mice had no viral neutralizing activity against heterologous virus. PMID- 12547602 TI - Liposomes and ISCOMs. AB - Liposomes and ISCOMs have a long history as vehicles for antigen delivery. Liposomes can carry both membrane associated antigens as well as water soluble molecules. Their physical properties are highly variable, depending on composition and manufacturing method. This allows optimised design for specific tasks (targeting, co-incorporation of adjuvants, etc.). ISCOMs already have a build-in adjuvant, Quillaja saponin, which is a structural part of the vehicle. In recent years, considerable progress has been achieved with respect to the use of better defined saponin. Clinical trials with ISCOMs are in progress and registered liposomal vaccines exist. Here, follows a brief overview on recent developments with emphasis on pharmaceutical aspects. PMID- 12547603 TI - Immunostimulating reconstituted influenza virosomes. AB - Immunostimulating reconstituted influenza virosomes (IRIVs) are a liposomal carrier system. Influenza hemagglutinin and neuraminidase are intercalated into the lipid bilayer and give the IRIVs their fusogenic activity. The system is already registered for human use and allows a specific targetting of antigens to a cellular or a humoral immune response. IRIVs are able to deliver proteins, RNA/DNA, peptides and mimetics to immunocompetent cells. PMID- 12547604 TI - Influenza virosomes: combining optimal presentation of hemagglutinin with immunopotentiating activity. PMID- 12547606 TI - Effect of priming on subsequent response to inactivated influenza vaccine. AB - Although shown to be a potent stimulator of serum antibody responses in animal models, the adjuvant immuno-stimulating complexes (ISCOMs) showed little adjuvant effect for inactivated influenza vaccines in a volunteer study. The result may be the non-comparability of the studies: animal studies were carried out chiefly in unprimed mice, while volunteers are mostly primed by previous infection and/or immunization. To test this, Balb/C mice were infected with influenza viruses or immunized with inactivated influenza vaccine, and subsequently given inactivated vaccine in saline or incorporated into ISCOMs. The serum in antibody responses was measured 1 month after immunization. The results confirm the adjuvant activity of ISCOM in unprimed mice, and show a marked reduction in adjuvant activity for primed mice. We argue that ISCOMs are important to prime the T cell response necessary for the serum antibody response to saline vaccine, but largely unnecessary where priming has been accomplished by prior exposure to influenza antigens. Further, the value of ISCOMs may lie in promoting antibody responses in unprimed subjects, and not in enhancing antibody titres. PMID- 12547605 TI - Influenza hemagglutinin peptides fused to interferon gamma and encapsulated in liposomes protects mice against influenza infection. AB - The immunogenicity of a peptide vaccine may be improved by fusing antigen to a cytokine and administering this chimeric protein in a particulate delivery system. We have investigated this using a vaccine comprising an immunodominant T cell epitope and a B cell epitope from influenza haemagglutinin (HATB) fused to interferon gamma and encapsulated in liposomes (HATB/IFN-gamma/lipo). Controls comprised groups receiving HATB/IFN-gamma mixed with liposomes, HATB incorporated in liposomes or heat inactivated PR8 influenza virus (HI PR8). IFN-gamma production in mice treated with HATB/IFN-gamma/lipo was significantly higher than in mice inoculated with either HATB/IFN-gamma mixed with liposomes or HATB incorporated in liposomes but less than HI PR8. Lung viral titres were significantly lower in mice treated with HATB/IFN-gamma/lipo compared with those treated with HATB/IFN-gamma mixed with liposomes. HI PR8-treated mice recorded a nil viral titre. There was no correlation between the level of antibody production and clearance of virus from the lungs. These data suggest that particulate delivery systems may be useful adjuncts to improve immune responses to chimeric proteins and to induce protection against disease. PMID- 12547607 TI - Intranasal vaccination with ISCOMATRIX adjuvanted influenza vaccine. AB - Mucosal delivery of inactivated vaccines that are able to elicit protective immune responses against respiratory diseases has been a long time goal of vaccinologists. Such vaccines would enable a more appropriate means of vaccination against respiratory diseases than those currently delivered by a parenteral route. The intranasal delivery of inactivated influenza vaccine plus the ISCOMATRIX (IMX) adjuvant, simply mixed together, was able to induce serum haemagglutination inhibition (HAI) titres in mice far superior to those obtained with unadjuvanted vaccine delivered subcutaneously. Furthermore, the IMX adjuvanted vaccine delivered intranasally induced mucosal IgA responses in the lung, nasal passages and large intestine, together with high levels of serum IgA. Intranasal delivery of IMX adjuvanted influenza vaccine in sheep gave antibody responses in both serum and nasal secretions that surpassed the levels obtained with unadjuvanted vaccine administered subcutaneously. These observations suggest that it may be possible to induce effective immunity to influenza in humans by intranasal vaccination with an IMX adjuvanted inactivated vaccine. PMID- 12547608 TI - Restored functional immunogenicity of purified meningococcal PorA by incorporation into liposomes. AB - The impact of the conformation, lipooligosaccharide (LOS)-depletion and the presentation form of outer membrane protein PorA from Neisseria meningitidis (PorA) subtype P1.7-2,4 on the immune response in mice was studied. Native PorA was purified from outer membrane vesicles (OMVs) derived from meningococci and reconstituted into liposomes. The conformation of PorA after purification from OMVs and reconstitution in liposomes was monitored by use of electrophoretic and spectroscopic techniques and compared with the conformation of PorA in outer membrane complexes (OMCs) and heat-denatured PorA. The antigenicity of the PorA formulations was measured by ELISA by using a bactericidal anti-P1.4 monoclonal antibody. Immunogenicity was determined in Balb/c mice. PorA-specific IgG, isotype distribution and bactericidal activity were measured after subcutaneous immunization. In all formulations except in heat-denatured OMVs, PorA was present as trimers. The lipooligosaccharide (LOS) content was reduced by 96% in the purified protein with respect to the original OMVs. The antigenicity of purified PorA (i.e. ELISA response) was substantially higher as compared to PorA in liposomes, OMVs or OMCs. The results of the immunogenicity studies showed that all formulations were able to induce comparable IgG titers. However, whereas the antibodies raised by OMVs were bactericidal, the antibodies elicited by immunization with purified PorA were unable to kill meningococci. Remarkably, the ability to induce bactericidal antibodies was fully recovered by incorporation of the purified PorA into liposomes, in the absence of other adjuvants, as compared to LOS-containing OMVs. PMID- 12547609 TI - Novel synthetic LPS receptor agonists boost systemic and mucosal antibody responses in mice. AB - Safe and cost-effective adjuvants are a critical requirement for subunit vaccine development. We report here the in vivo activity of a series of fully synthetic LPS receptor agonists that have been shown to activate NF-kappaB signaling through the Toll-like receptor 4 (TLR4). These compounds boost antibody responses to protein antigens when coadministered at microgram doses in mice. At these dosage levels no adverse effects are observed. Antibody responses are largely IgG1, with enhanced IgG2a, and down-regulated IgE as compared to alum adjuvanted immunization. Stimulation of Th1 is confirmed by enhanced gamma-interferon production after in vitro antigen restimulation of spleen cells from mice immunized with the synthetic adjuvants. The adjuvants are active by both subcutaneous and intranasal routes of vaccine administration, and in the latter case can amplify both serum IgG and serum and mucosal IgA responses. The compounds must be administered at the same site with antigen to boost anti vaccine antibody. These fully synthetic ligands of the innate immune system offer the potential for use as effective, safe, and nonbiologically-derived adjuvants. PMID- 12547610 TI - Adjuvant activity of peptidoglycan monomer and its metabolic products. AB - Peptidoglycan monomer (PGM) is a natural compound of bacterial origin. It is a non-toxic, non-pyrogenic, water-soluble immunostimulator potentiating humoral immune response to ovalbumin (OVA) in mice. It is fast degraded and its metabolic products-the pentapeptide (PP) and the disaccharide (DS)-are excreted from the mammalian organism upon parenteral administration. The present study investigates: (a). whether PGM could influence the long-living memory generation; (b). whether metabolic products retain adjuvant properties of the parent compound and contribute to its adjuvanticity. We report now that mice immunised twice with OVA+PGM had significantly higher anti-OVA IgG levels upon challenge with antigen alone 6 months later in comparison to control group immunised with OVA only. PP and DS were prepared enzymatically in vitro as apyrogenic and chemically pure compounds. When mice were immunised with OVA plus PP and DS, respectively, the level of anti-OVA IgGs in sera was not higher than in mice immunised with OVA alone, while PGM raised the level of specific antibodies. Results implicate that the adjuvant active molecule, capable of enhancing long-living memory generation, is PGM itself, and none of its metabolic products. PMID- 12547611 TI - Evaluation of adjuvants for protein vaccines against tuberculosis in guinea pigs. AB - Subunit vaccines against tuberculosis show promise but require administration with adjuvants to stimulate relevant immune responses for protection. Guinea pigs are the model of choice for evaluating protective immunity to aerogenic challenge with virulent mycobacteria, but few studies have been undertaken to identify suitable adjuvants for vaccine screening in this species. Here, we compare the efficacy of several adjuvants to induce T cell responses to culture filtrate protein in guinea pigs. We report that of several adjuvants tested, the most promising was CpG ODN formulated in an aqueous emulsion. This adjuvant induced type 1 T cell responses equivalent to that of FIA, as measured by delayed-type hypersensitivity reactions (DTH), antigen-specific T cell proliferation and antigen-specific IgG1 and IgG2 responses. These data demonstrate the potential for CpG motif based adjuvants for use in TB vaccine screening in guinea pigs, and other diseases where a type 1 T cell response is required. PMID- 12547612 TI - Development of short non-CpG phosphodiester oligonucleotides as immune stimulatory agents. AB - We have previously reported that DNA isolated from Mycobacterium phlei (M. phlei) stimulates the synthesis of cytokines by monocytes and macrophages independently of the presence of unmethylated CpG motifs. Oligonucleotides as small as five to six bases isolated from M. phlei DNA have been found to induce cytokine synthesis. In the present study, we have investigated the potential for such CpG lacking DNA to act as an immune stimulant. A series of six base length phosphodiester oligonucleotides derived from the genome of M. phlei were synthesised and tested for their ability to induce the synthesis of cytokines by murine, non-human primate (rhesus macaques and chimpanzee) and human peripheral blood mononuclear cells. The results show that phosphodiester oligonucleotides with a 5'GGGxGG3' sequence where x is A, C, G or T have the ability to induce the synthesis of IL-1beta, IL-6, IL10 or IL-12 by non-human primate and human PBMC, murine cells being unresponsive. The phosphodiester 5'GGGxGG3' oligonucleotides were shown to be stable in human serum, with a half-life of approximately 72 h. The addition of aluminium hydroxide to these 5'GGGxGG3' oligonucleotides potentiated, in a concentration-dependent manner, the synthesis of IL-12 by human peripheral blood mononuclear cells. These phosphodiester six base length non-CpG motif oligonucleotides may have potential as immunopotentiators for vaccines. PMID- 12547613 TI - Immunostimulatory DNA as adjuvant: efficacy of phosphodiester CpG oligonucleotides is enhanced by 3' sequence modifications. AB - The immunostimulatory activity of oligonucleotides containing CpG motifs is well established and represents the basis for an effective vaccine adjuvant. For use in vivo CpG ODN have to be protected from the attack of nucleases to ensure sustained effectiveness. This is usually accomplished by phosphothioate (PTO) modifications of the ODN's backbone. However, PTO modification may induce undesired effects. We, therefore, have attempted to enhance the immunostimulatory activity of CpG phosphodiester ODN by supplementation of the ODN's sequence. We report here, that addition of poly-guanosine runs to the 3'-end of a CpG-PO ODN conveys immunostimulatory activity to CpG-PO ODN in vivo and in vitro. Guanosine rich PO ODN thus might be an alternative approach to develop effective and safe vaccine adjuvants. PMID- 12547614 TI - A non-toxic lectin for antigen delivery of plant-based mucosal vaccines. AB - RicinB, the non-toxic galactose/N-acetylgalactosamine-binding subunit of ricin, was fused to a model antigen, green fluorescent protein (GFP), and expressed in tobacco plants and hairy root cultures to test for utility in mucosal vaccine delivery/adjuvancy. The fusion protein retained both GFP fluorescence and galactose/galactosamine-binding activity. Intranasal immunization of mice with galactosamine-affinity purified ricinB:GFP recovered from tobacco root cultures triggered significant increases in GFP-specific serum IgGs. This strong humoral response was comparable to that observed following GFP immunization with cholera toxin adjuvant. GFP at the same concentrations but without an adjuvant was non immunogenic. Induction of higher levels of IgG(1) than IgG(2a) following ricinB:GFP immunization suggested the presence of a Th2 response. Serum and fecal anti-GFP IgA were also induced by immunization with ricinB:GFP. Our data suggest that ricinB can be used as an adjuvant and antigen carrier to the mucosa and is efficient in eliciting systemic and mucosal immune responses. PMID- 12547615 TI - Investigation of several selected adjuvants regarding their efficacy and side effects for the production of a vaccine for parakeets to prevent a disease caused by a paramyxovirus type 3. AB - The infection with paramyxovirus type 3 (PMV-3) of parakeets can lead to severe illness in small psittacines (Neophema spp. and other parakeets) as well as in passerines (finches). The disease is characterized by acute or chronical pancreatitis and central nervous symptoms, such as torticollis as well as walking in circles and by high mortality rates in the affected flocks. As there is no licensed vaccine for psittacines available to prevent this disease. The aim of the following study was to find a suitable vaccine formulation for parakeets with inactivated PMV-3 and a well-tolerated and effective adjuvant. Seven adjuvants have been examined in ovo and in vivo regarding their efficacy and side effects. In these investigations, the classical Freund's complete Adjuvant (FCA) and Freund's incomplete Adjuvant (FICA) and Alhydrogel, but also the more recent developments TiterMax Gold, Specol, Gerbu Adjuvant 100, and Diluvac Forte have been used. Regarding its efficacy and side effects, the vaccine formulation "PMV 3/Specol" has been evaluated positively and can be recommended for the production of a PMV-3 vaccine for parakeets. PMID- 12547617 TI - Retrovirus testing: the mainstay remains. PMID- 12547616 TI - Reduced faecal excretion of F4+-E coli by the intramuscular immunisation of suckling piglets by the addition of 1alpha,25-dihydroxyvitamin D3 or CpG oligodeoxynucleotides. AB - In this study it was analysed whether intramuscular (IM) immunisation of piglets with F4 during the suckling period could protect against oral challenge with F4(+)-Escherichia coli and whether addition of 1alpha,25(OH)(2)D(3) or CpG-ODN could improve this protection.F4-seronegative F4-receptor positive pigs were divided into four groups of five pigs each. The pigs were intramuscularly injected with F4 fimbriae only or supplemented with 1alpha,25(OH)(2)D(3) (D(3) group) or CpG-ODN (CpG-group). The control group received PBS in IFA. Seven days after the second immunisation, all pigs were intragastrically inoculated with 1 x 10(10) CFU of F4(+)-E. coli. All F4-injected groups, showed a reduced faecal excretion of F4(+)-E. coli. However, this reduction was only statistically significant in the D(3)-group 2 days post challenge. Pigs in the latter group showed a secondary antibody response upon challenge, indicating that F4-primed memory B-cells were present in the gut-associated lymphoid tissues at that moment.CpG-ODN, on the other hand, did not enhance the F4-specific antibody response. However, CpG-ODN significantly increased the F4-specific as well as mitogen-induced proliferation of peripheral blood monomorphonuclear cells indicating a direct or indirect overall effect on T-lymphocytes. In conclusion, supplementation with 1alpha,25(OH)(2)D(3) or CpG-ODN improved protection against an F4(+)-E. coli infection. This protection was most obvious for 1alpha,25(OH)(2)D(3) and indicates its potential use in veterinary vaccines against enteropathogens. PMID- 12547618 TI - Clinicopathological findings in five cats with paw calcification. AB - This retrospective study describes the clinicopathological findings in five cats with soft tissue mineralisation of interdigital spaces and footpads. Paw disease was the reason for veterinary consultation in three out of five cats. All cats had laboratory findings suggestive of renal failure and high solubility product [calciumxphosphorus]. In all cases, cytological examination of paw lesions was suggestive of calcinosis. The results of our study agree with two previous case reports of paw calcification in the cat, suggesting a metastatic pathogenesis and a correlation between paw mineralisation and renal failure. PMID- 12547619 TI - Panniculitis, due to Mycobacterium smegmatis, in two Finnish cats. AB - Pyogranulomatous panniculitis due to infection by Mycobacterium smegmatis was diagnosed in two cats in Finland, a country with a rather cold climate. The diagnosis was confirmed by sequencing of the 16S rRNA gene, which gave a perfect match with the M smegmatis strain ATCC 19420. Gene sequencing makes it possible to distinguish M smegmatis from closely related mycobacteria such as M goodii sp.nov. Diagnosing this entity seems to be a question of having a high index of suspicion. The appearance of the disease as well as sampling is described in detail. In our first case an initial erroneous diagnosis of Nocardia species considerably delayed our arriving at the right diagnosis. The first patient has now been followed for more than 7 years. Her disease is chronic, but she is not systemically affected. Several antimicrobials were tried. Probable side effects of enrofloxacin medication are described. PMID- 12547620 TI - Feline cystic thymoma: a clinicopathologic, immunohistologic, and electron microscopic study of 14 cases. AB - Cystic thymoma was diagnosed in 14 cats in a period of 6 years. The most common clinical sign was laboured breathing. The tumours were characterized by various sized cystic spaces with central vessels. The epithelial cells varied from oval to spindle to polygonal cells enclosing cystic spaces or in pure epithelial cell components. The nuclei of the neoplastic cells had scattered chromatin and small nucleoli. The cytoplasm was pale eosinophilic. The concentration of mature lymphocytes varied from area to area with rare germinal centres. Immunohistochemically, the epithelial cells stained only with AE(1)/AE(3). The central vessels were positive for vimentin, smooth muscle actin, and factor VIII antigen. Electron microscopy revealed that the cyst walls were lined by epithelial cells that were joined by desmosomes, and the walls were well separated from the cystic spaces by a well-defined basement membrane. The neoplastic epithelial cells contained scattered tonofilaments. Three of the cats had metastasis to the lymph nodes and lungs. Two novel cases of ectopic cystic thymoma have also been described. Results of this study reveal that cystic thymoma is uncommon in cats, and that the histomorphologic, immunohistochemical, and electron microscopic features are similar to those of cystic thymoma in humans. PMID- 12547621 TI - Opportunistic mycobacterial granuloma in a cat associated with a member of the Mycobacterium terrae complex. AB - An 18-month-old domestic short-haired neutered male cat presented with a nodular dermal thickening on a digit. Biopsy demonstrated pyogranulomatous inflammation with moderately frequent acid-fast bacilli. A member of theMycobacterium terrae complex was isolated. There was no evidence of systemic involvement. Treatment was initiated with enrofloxacin, rifampicin and clarithromycin. After 2 months there was no longer any clinically apparent dermal thickening. Treatment was continued for a further 3 months using enrofloxacin and rifampicin. PMID- 12547622 TI - Caudal mediastinal abscess due to a grass awn (Hordeum spp) in a cat. AB - A two-and-a-half year-old male, domestic shorthaired cat was presented with a history of chronic expiratory dyspnoea, dysphagia and progressive weight loss. Thoracic radiography revealed a caudal mediastinal mass. Surgical exploration confirmed the presence of an abscess in the caudal mediastinum. Despite the good prognosis, euthanasia was performed at the owner's request and post-mortem examination revealed an abscess that had been caused by a grass awn located within. PMID- 12547623 TI - The rainbow trout classical MHC class I molecule Onmy-UBA*501 is expressed in similar cell types as mammalian classical MHC class I molecules. AB - Onmy-UBA is a polymorphic classical major histocompatibility (MHC) class I locus in rainbow trout (Oncorhynchus mykiss). A common allomorph is Onmy-UBA*501, which has been detected in several wildtype strains, in the clonal homozygous rainbow trout C25 and, in the current study, in the rainbow trout gonad cell line RTG-2. The extracellular domain of this allomorph was expressed in E. coli and a murine monoclonal antibody designated H9 was generated against the recombinant protein. In Western blot analysis Mab H9 specifically recognised an n-glycosylated protein of 45 kDa in leucocytes and erythrocytes of C25 fish and in RTG-2 cells. The level of Onmy-UBA*501 expression in erythrocytes was very low. Immunocytochemistry of isolated cells indicated expression in lymphocytes, macrophages, neutrophils, erythrocytes, RTG-2 cells and Onmy-UBA *501 transfected CHO cells, but not in untransfected CHO cells. Immunohistochemistry using frozen sections of C25 fish indicated that Onmy-UBA*501 expression is strong in the lymphoid organs (thymus, head kidney and spleen) and in the epithelia and endothelia of several organs. No significant expression was observed in muscle fibres, hepatocytes or neurons. These observations demonstrate that in jawed fish, the lowest phylogenetic group possessing an MHC system, the classical MHC class I molecules are expressed in similar cell types as in higher vertebrates. PMID- 12547624 TI - The influence of temperature and dose on antibacterial peptide response against lipopolysaccharide in the blue mussel, Mytilus edulis. AB - Blue mussels (Mytilus edulis) were inoculated with two different doses of lipopolysaccharides (LPS) or phosphate-saline (PS) buffer under different temperature conditions (6 and 20 degrees C). The activity of the antibacterial peptide fraction, purified through reverse phase chromatography from mussel haemolyph, was compared at different time intervals after the inoculation. The activity was determined as the minimal peptide concentration that inhibited growth of the Gram-negative bacteria Escherichia coli D21, by using radial diffusion assay. The antibacterial activity for mussels inoculated with LPS changed over time, both at 6 and 20 degrees C, but those inoculated with PS buffer did not. The response was enhanced within a time course of 3h. The higher temperature did increase the inhibitory activity and made the mussel respond at an earlier stage, in comparison to that at 6 degrees C. At 20 degrees C, mussels inoculated with 10 microg of LPS responded faster than those inoculated with 0.1 microg of LPS. In addition, cytotoxic effects of LPS on mussel haemocytes were investigated in vitro, using a colorimetric assay. The survival index (SI%) for haemocytes decreased with 76% at 6 degrees C but increased with 100% at 20 degrees C, irrespective of the dose of LPS. This indicated that LPS did not influence the viability of the haemocytes but the high temperature increased their metabolic state. Likely, antibacterial response was provoked by LPS in a dose-dependent manner and favoured by higher metabolic state of the haemocytes, elicited at higher temperature. These results provide important considerations for variability in the internal defence of mussels and consequently, also the retention of viable human pathogens in mussels. PMID- 12547625 TI - In vivo effects of beta-glucan and LPS on regulation of lysozyme activity and mRNA expression in Atlantic salmon (Salmo salar L.). AB - The present study was undertaken to compare the effects of intraperitoneally injected bacterial lipopolysaccharide (LPS) and yeast beta-glucan on lysozyme activity in Atlantic salmon, and to explore what organ(s) are responsible for the increase in plasma lysozyme activity induced by the compounds. The results indicated that LPS stimulates plasma lysozyme activity at least as efficiently as beta-glucan. The lysozyme gene was shown to be transcribed in head kidney, spleen, liver and intestine, and accumulation of transcript was demonstrated in response to both beta-glucan and LPS in all of these organs. Intracellular lysozyme activity was detected in the same organs and in isolated blood polymorphonuclear cells (PMN) and lymphocytes. Increased lysozyme activity in response to both beta-glucan and LPS was demonstrated in blood PMN and cells isolated from head kidney and intestine. In spleen and liver on the other hand, there was no increase in lysozyme activity in response to the stimulants. Based on previous work and the present results it is suggested that plasma lysozyme induced by LPS and beta-glucan originate from macrophages in the different organs. The head kidney is likely to be the main supplier of plasma lysozyme considering its high contents of macrophages. This work supports the notion that microbial compounds containing phylogenetically conserved structures (beta-glucan and LPS) are able to stimulate the non-specific defence of animals against infection by enhancing the lysozyme expression. PMID- 12547626 TI - Cloning, sequence and variability analysis of expressed immunoglobulin light chain genes from yellowtail Seriola quinqueradiata. AB - The cDNA clones encoding immunoglobulin (Ig) light (L) chain variable (V) region associated with constant (C) region were isolated from yellowtail (Seriola quinqueradiata) kidney by expressed sequence tag analysis (accession numbers: AB062619-AB062668, AB064322). The sequences of both VL and CL region contain well conserved cysteine residues important for intra- and inter-domain interaction in mammals. Comparisons of the amino acid sequence of the CL domain with those of other species showed a high degree of similarity, with 88.3%, 59.8%, and 60.6% to those of wolf fish (Anarhichas lupus), rainbow trout IgL I isotype (Oncorhynchus mykiss) and channel catfish G isotype (Ictalurus punctatus), respectively. Multiple sequence alignments of the CL domain with those of higher vertebrates, however, did not readily allow it to be classified as kappa or lambda isotypes. Furthermore, the pI, hydrophobicity and variability of yellowtail VL regions were studied in 65 cDNA clones and the diversity was observed in CDR1, CDR2 and CDR3 regions. PMID- 12547627 TI - Limited diversity of the immunoglobulin heavy chain variable domain of the emerald rockcod Trematomus bernacchii. AB - To investigate the diversity of the immunoglobulin heavy chain variable domain of the cold adapted teleost Trematomus bernacchii, 45 cDNA clones, containing complete or partial sequences of rearranged VH/D/JH segments, were analysed. Clones were isolated from a spleen library constructed by 5' RACE or from an expression library previously constructed and immunoscreened with rabbit anti- T. bernacchii Ig heavy chain antibodies. VH sequences shared, on average, 79.9% nucleotide identity and defined only two gene families referred to as Trbe VH I and Trbe VH II, the latter comprising 89% of the VH sequences analysed in this study. A Southern blot analysis, performed with family specific probes, revealed that there are at least 25 genomic VH genes. A phylogenetic tree showed that Trbe VH I clustered with VH genes belonging to group D and Trbe VH II with those of group C. Four putative distinct D segments were found to contribute to the diversity of CDR3, which showed a high glycine content. The Shannon analysis revealed that FRs are very highly conserved. Of CDRs, CDR2 exhibits a mean entropy value higher than CDR1, contributing to variability in a significant manner. Moreover, eight distinct JH segments were identified. These findings provide several clues suggesting a limited diversity of the VH genes in the Antarctic teleost T. bernacchii. PMID- 12547628 TI - Induction of cytochrome P-450 activity in individual Chironomus riparius Meigen larvae exposed to xenobiotics. AB - Cytochrome P450 activity in individual Chironomus riparius larvae was measured using a microtiter plate adaptation of the ethoxyresorufin-O-deethylase (EROD) assay. The sensitivity of this biomarker was tested by exposing larvae to phenobarbital (0.5 and 1.0 mM) and permethrin (1 and 10 microg/g). Both chemicals induced EROD activity in C. riparius larvae by up to 1.58-fold with PB and 2.47 fold with permethrin. EROD induction was more pronounced after 48 h. The initially high EROD activity in the controls suggested that P450s are induced by stress. Feeding levels prior to exposure also had a significant effect on EROD activity. EROD activity compared to the control was highest when larvae were fed double the normal ration. These results indicate that EROD activity in individual C. riparius may be a useful biomarker to add to a suite of biomarkers for the detection of freshwater pollution. PMID- 12547629 TI - Naphthalene-induced differential tissue damage association with circulating fish phagocyte induction. AB - The effect of naphthalene on phagocytes and associated abnormal cellular activities was studied in Anguilla anguilla L. Fish were exposed to sublethal concentrations (0.1, 0.3, 0.9, 2.7 microM) of naphthalene for 8, 16, 24, 48, and 72 h. Gill, head kidney, and peritoneum phagocyte respiratory burst activity (RBA) was measured by nitroblue tetrazolium reduction assay, whereas lipid peroxidation in fish gill, kidney, and liver was measured by thiobarbituric acid reaction substance. A significant increase (P < 0.05-P < 0.001) in total cell count (TCC) of phagocytes (TCCPH) isolated from gill, head kidney, and peritoneum was observed after 8 h of naphthalene exposure and persisting at 16 h exposure. However, long-term exposures (24, 48, and 72 h) induced a significant (P < 0.05-P < 0.001) decrease in TCC at all the concentration levels. Naphthalene exposure caused a significant (P < 0.05-P < 0.001) RBA induction after 8 and 16 h in gill, head kidney, and peritoneal phagocytes, which consequently resulted in a significant (P < 0.01 and P < 0.001) peroxidative tissue damage increase measured as lipid peroxidation (LPO) in gill, kidney, and liver at the same time intervals. Considering TCCPH, RBA, and LPO, the most affected tissues were gill and kidney after 8 and 16 h exposure to all naphthalene concentrations. RBA was significantly decreased after 24, 48, and 72 h of exposure (P < 0.05-P < 0.001), whereas peroxidative damage increased significantly (P < 0.05-P < 0.001) and persisted in long-term exposures (72 h) at all concentration levels in gill and kidney. Liver short-term exposure (8 and 16 h) to all naphthalene concentrations did not alter LPO activity. Long-term exposures (48 and 72 h) caused a significant (P < 0.01 and P < 0.001) LPO increase, which was more pronounced at 72 h. The results demonstrate that the activation pattern of RBA was corroborated by the extent of phagocyte-induced peroxidative damage in the tissues, as demonstrated by a significant increase of circulating phagocytes. However, the route of exposure and mode of entry of a pollutant may affect the activation pattern of circulating fish phagocytes. It is proposed that measurement of phagocyte-induced reactive oxygen species and their association with peroxidative damage in fish tissues may prove to be useful in biomonitoring fish exposure to aquatic pollutants. PMID- 12547630 TI - Effects of estrogenic and antiandrogenic compounds on the testis structure of the adult guppy (Poecilia reticulata). AB - Sexually mature male guppies (Poecilia reticulata) were exposed to 17beta estradiol, 4-tert-octylphenol, bisphenol A, flutamide, p,p'-DDE, or vinclozolin to determine the effects of these estrogenic or antiandrogenic substances on the testis structure. Flutamide, p,p'-DDE, 4-tert-octylphenol, and bisphenol A exposure caused similar effects. These included a reduced number of spermatogenetic cysts and an increased number of spermatozeugmata in the ducts. These effects are indicative of a blocked spermatogonial mitosis. 17beta Estradiol produced effects different from those of the other compounds studied. The effects were seen as an increased number of hypertrophied Sertoli cells/efferent duct cells. The mammalian antiandrogen vinclozolin had no observable effects on the testis structure when given in doses of 1, 10, or 100 microg/mg food. PMID- 12547631 TI - Evaluation of genotoxicity of PCP and 2,4-D by micronucleus test in freshwater fish Channa punctatus. AB - An in vivo study on the genotoxic effects of pentachlorophenol (PCP) and 2,4 dichlorophenoxyacetic acid (2,4-D) was carried out with freshwater air-breathing fish Channa punctatus. The fish were exposed to three sublethal doses of PCP (0.2, 0.4, and 0.6 ppm) and 2,4-D (25, 50, and 75 ppm) by medium treatment. Micronucleated erythrocytes were sampled at intervals of 48, 72, and 96 h post treatment. Student's t test revealed significant increases in micronucleus (MN) frequency. Maximum incidence of MN was recorded at highest concentrations and durations for both chemicals. A time- and dose-dependent response of the MN frequency for both chemicals was confirmed. PCP-treated groups had 8-10 MN per cell, specifically at higher concentrations and duration. Echinocytes were recorded at higher concentrations of 2,4-D. The study confirmed the genotoxicity of PCP and 2,4-D in this organism. PCP was found to be more toxic than 2,4-D in terms of MN induction. This assay has further potential for detecting genotoxic compounds in aquatic ecosystems. PMID- 12547632 TI - Acute toxicity effects of tributyltin chloride and triphenyltin chloride on gilthead seabream, Sparus aurata L., embryos. AB - Acute toxicity studies, using two organotin compounds, tributyltin chloride (TBTCl) and triphenyltin chloride (TPhTCl), were performed on gilthead seabream, Sparus aurata, fertilized eggs. The tests were static and run for 24 h (temperature = 19 +/- 1 degrees C, salinity = 39.5 g/L). Three replicate egg groups were incubated in solutions containing different concentrations of the toxicants. The (24h)LC(50) value for TBTCl was 28.3 microg/L and for TPhTCl was 34.2 microg/L, when the embryos were at the morula and late blastula stage at the start of the experiment. The value for TBTCl was 38.6 microg/L, when the embryos were at the gastrula stage at the start of the experiment. The results reconfirm the expected toxicity of TBT to this euryhaline fish species and introduce TPhT as a toxicant of potentially similar impact on teleosts. PMID- 12547633 TI - Studies of peripheral benzodiazepine receptors in mussels: comparison between a polluted and a nonpolluted site. AB - The aim of this study was to investigate the peripheral benzodiazepine receptors in soft tissue membranes of the mussel Mytilus galloprovincialis from both polluted and nonpolluted seawater populations, using a radioligand specific for this receptor, [3H]PK11195. Mussels were dissected into four body parts--mantle, gills, digestive gland, and muscles-to determine the distribution of tissue specific peripheral benzodiazepine receptors (PBRs). The specific binding was saturable and reversible. A statistically significant increase (muscle, 537% and mantle, 201%, as absolute percentages) in the maximal number of binding sites (B(max)) was found in mussels from the polluted site, compared with mussels from the nonpolluted site. By contrast, the value of the dissociation constant (K(d)) at equilibrium does not show a statistically significant variation between the two groups. In competitive experiments of the compounds clonazepam, flumazenil, flunitrazepam, Ro5-4864, PK11195, and protoporphyrin IX, only PK11195 and protoporphyrin IX displaced [3H]PK11195 specifically bound to soft tissue membranes, revealing that the binding sites of peripheral benzodiazepine receptors of mussels have pharmacological properties comparable to those of low vertebrates such as trout. M. galloprovincialis was also tested as an indicator of heavy metal exposure, and metal accumulation in the digestive gland was measured by atomic absorption spectrophotometry (AAS). The contents of Pb, Mn, and Zn in mussels collected off the polluted site were higher than those in mussels from the nonpolluted site. These data suggest that PBRs are present in the soft tissues of the mussel M. galloprovincialis. Here we report preliminary evidence of biochemical alterations in mussels from the polluted site. PMID- 12547634 TI - Use of atropine-treated Daphnia magna survival for detection of environmental contamination by acetylcholinesterase inhibitors. AB - The toxicity of cholinesterase-inhibiting compounds (e.g., carbamates and organophosphates) is due to a decrease in acetylcholine metabolism, which results in a continuous stimulation of cholinergic receptors (muscarinic and nicotinic) that can be fatal. The goal of this study was to evaluate the protective effect of atropine (muscarinic receptor antagonist) against paraoxon-induced toxicity to Daphnia magna using its survival rate for the detection of environmental contamination by cholinesterase-inhibiting compounds. As expected, paraoxon was lethal to D. magna in a concentration-dependent manner. Noteworthy, the pretreatment of these organisms with atropine dramatically increased their survival against paraoxon. These results indicate that muscarinic stimulation plays an important role in paraoxon-induced lethality in D. magna. Therefore, simply by using the survival of atropine-treated and nontreated D. magna, water contamination by cholinesterase-inhibiting compounds may be rapidly and specifically detected. PMID- 12547635 TI - Distribution of petroleum hydrocarbon in sediment from coastal area receiving industrial effluents in Kuwait. AB - Sediment samples from the coastal area facing Shuaiba industrial area (15 x 1.5 km(2)) were examined for contamination with petroleum hydrocarbons. Aliphatic hydrocarbons in sediment extracts were characterized largely by unresolved complex mixture from nC(22)-nC(33). The sediment samples from the upstream area facing Mina Al-Ahmadi refinery to Shuaiba Harbor were heavily polluted with polycyclic aromatic hydrocarbons (PAHs) possibly due to the higher depth in the area created for a navigational channel to the harbor. PAHs at these locations exceeded the probable effect levels with reference to sediment quality guidelines. PMID- 12547636 TI - Effects of different plant protection programs on soil microbes. AB - The aim of this study was to assess the effects of two pesticide regimens (conventional full insurance compared with supervised and reduced pesticide use) and two cultivation techniques (conventional tillage and fertilization compared with no tillage and lower fertilization) on soil microbes in a field study, and to evaluate the results with laboratory tests. The herbicides used were chlorsulfuron, MCPA, and bentazone; the fungicides carboxin-imazalin and propiconazole; and the insecticides dimethoate and pirimicarb. In the field studies, the effects on microbial biota were assessed by measuring biomass (ATP content) and microbial activities related to carbon and nitrogen metabolism (soil respiration, nitrification potential). Potential harmful effects of commercial formulations of the same pesticides on microbes were studied in the laboratory with two bacterial toxicity tests (Pseudomonas putida growth inhibition and Vibrio fischeri luminescence inhibition tests) and with a soil respiration inhibition test. Bioavailability of the pesticides in the soil was assessed by a solid-phase modification of the luminescent bacteria test. In the field studies the microbial activities and biomass followed the weather conditions during the growing season, and significant effects of pesticide treatments on microbial processes were not observed. In the laboratory studies the toxicity of certain pesticides was clearly detected by bacterial toxicity tests. However, in the soil respiration inhibition assessment with soil similar to that used in the field trial, inhibition was observed only at unrealistically high concentrations. This could be due to the limited bioavailability of the pesticides in soil of high clay and organic carbon content. PMID- 12547637 TI - Mobility and toxicity of metals in sandy sediments deposited on land. AB - A times series of laboratory experiments were conducted to investigate the effect of land deposition of contaminated sediments on the bioavailability and mobility of metals. Four sandy sediments were sampled at sites expected to have elevated levels of cadmium and zinc. The physical and chemical characteristics and ecotoxicity of sediments, pore waters, and leachates were evaluated after periods ranging from 1 to 45 days of land deposition. Cd and Zn retardation and leaching potential were calculated and this simulation gave good predictions of subsequently observed Cd and Zn mobility. The mobility and leaching of Cd and Zn in the sediments increased with decreasing pH and with decreasing content of organic matter. During the deposition an increase in sediment toxicity to plants and an increase in eluate toxicity to invertebrates were observed. A high rate of water flow through the sediment resulted in a lower toxicity enhancement of the sediments and a higher toxicity enhancement of the eluates. This result suggests that water flow through the sediment reduces the actual toxicity of the upper layer of deposited sediment but at the same time intensifies the risk of groundwater contamination. PMID- 12547638 TI - Application of toxicity tests into discharges of the pulp-paper industry in Turkey. AB - The aim of this study was to investigate the acute toxicity of pulp-paper industry wastewater using traditional and enrichment toxicity tests and to emphasize the importance of toxicity tests in wastewater discharge regulations. Enrichment toxicity tests are novel applications and give an idea of whether there is potential toxicity or growth-limiting and -stimulating conditions. Different organisms were used such as bacteria (floc and coliform bacteria), algae (Chlorella sp.), protozoa (Vorticella sp.), and fish (Lepistes sp.) to represent four trophic levels. Furthermore, chemical oxygen demand (COD) fractionation results were compared with these tests to assess the effect of COD subcategories on the determination of possible toxicity. The pulp-paper industry results revealed acute toxicity to at least two organisms in 6 of 20 effluent samples. The toxicity test results were assessed with chemical analyses such as COD, biochemical oxygen demand (BOD), color, absorbable organic halogen (AOXs), and phenol. It was observed that the toxicity of the effluents could not be explained by using physicochemical analyses in four cases for the pulp-paper industry. The results clearly indicate that bioassay tests provide additional information on the toxicity potential of industrial discharges and effluents. PMID- 12547639 TI - The toxicity of cationic surfactants in four bioassays. AB - The purpose of this study was to investigate the toxicity of 15 quaternary ammonium compounds (QACs) in a battery of four bioassays comprising the bacterium Vibrio fischeri, two ciliated protozoa Spirostomum ambiguum and Tetrahymena thermophia, and the anostracean crustacean Artemia franciscana. The compounds were prepared by Professor Pernak's group at Poznan University of Technology (Poland). The toxicity of the test compounds was very high, with EC(LC)(50) values varying from 0.11 to 70 micromol/L. Microtox was the most sensitive bioassay, while the crustacean test was the least sensitive. Among the protozoa T. thermophila was 5-30 times less sensitive than S. ambiguum. The toxicity of the QACs depended on their structure, but no simple correlation was found for all the bioassays applied. PMID- 12547640 TI - Evaluation of an in vitro hsp70 induction test for toxicity assessment of complex mixtures: comparison with chemical analyses and ecotoxicity tests. AB - The aim of this study was to assess the potential of a human cell line containing the hsp70 promoter linked to the chloramphenicol acetyltransferase reporter gene in evaluating the toxic potential of complex mixtures. Cells were exposed to eluates of industrial wastes and the cellular responses were compared with the metal contents of the samples and with standardized aquatic (microalgal growth inhibition, daphnia Immobilization, bacterial luminescence inhibition, Ceriodaphnia dubia reproduction inhibition) and terrestrial (earthworm lethality, plant growth inhibition) tests. The hsp70 promoter was significantly induced by 11 of 14 samples, with different dose-response patterns. Significant correlations of in vitro induction potency with aquatic ecotoxicity, especially with chronic tests, and with the metal contents of the samples were observed. Our study provides new information on the relevance of hsp70 gene induction as a criterion of toxicity and suggests its usefulness for the detection of toxicity associated with metallic pollution in complex mixtures. PMID- 12547641 TI - The toxicity of fluoranthene to Hyalella azteca in sediment and water-only exposures under varying light spectra. AB - In the US Environmental Protection Agency methods for sediment toxicity testing, the light regimen is specified as a 16:8 light dark cycle with 500-1000 lx. The potential for photoinduced toxic effects from this requirement is evaluated. Hyalella azteca were exposed to fluoranthene in both water only and sediment to examine the impact of light spectra on the toxicity of fluoranthene. The light sources included gold fluorescent light (lambda > 500 nm), cool white fluorescent light, and UV-enhanced fluorescent light. Toxicity was determined as mortality after 10 days of exposure. The extent of mortality was determined both as LC(50) and LR(50) (median lethal body residue). In water-only exposures, the toxicity of fluoranthene was greatest under the UV-enhanced spectra, followed by fluorescent light, and least toxic under the gold light. Both the LC(50) and LR(50) values exhibited the same pattern. The toxicity under gold light gave an LR(50) of 0.81 mmol kg(-1) (0.82-0.79, 95% CI) similar to values expected for the acute toxicity of nonpolar narcotic (anesthetic) compounds. The LR(50) values under the other two light sources were substantially lower, 4 and 58 times lower for the fluorescent and UV-enhanced exposures, respectively. In sediment, toxicity was not significantly affected by the light source. Toxicity occurred only when the body residue concentration approached that of the LR(50) under gold light from the water-only exposures. Thus, H. azteca were significantly protected from the light by burrowing into the sediment. PMID- 12547642 TI - An NMR study of alterations in [1-13C]glucose metabolism in C6 glioma cells by gliotoxic amino acids. AB - A series of glutamate analogues, known as gliotoxins, are toxic to astrocytes in culture, and are inhibitors or substrates of high affinity sodium-dependent glutamate transporters. The mechanisms by which these gliotoxins cause toxicity are not fully understood. The effects of a series of gliotoxic amino acids (L alpha-aminoadipate, L-serine-O-sulphate, D-aspartate and L-cysteate) on metabolism of [1-13C]glucose were examined in C6 glioma cells using 13C nuclear magnetic resonance (NMR) spectroscopy. The cells were preincubated in the presence of sub toxic concentrations of each gliotoxin (400 micromol/l) for 20 h. This was followed by incubation (4 h) with [1-13C]glucose (5.5 mmol/l) in the presence and absence of each gliotoxin. The incorporation of 13C label into the observed metabolites was analysed. Following preincubation with L-alpha aminoadipate, D-aspartate, and L-serine-O-sulphate there was a significant decrease in the incorporation of 13C label into glutamate, alanine and lactate from [1-13C]glucose. In the presence of L-cysteate production of labelled glutamate was decreased, while there was no significant effect on the concentrations of labelled lactate and alanine. There was no change in the quantity of LDH released into the medium after incubation of the cells with any of the gliotoxins. Overall these results indicate that the presence of gliotoxins profoundly alters the flux of glucose to lactate, alanine, aspartate and glutamate. PMID- 12547644 TI - Distribution and expression of A1 adenosine receptors, adenosine deaminase and adenosine deaminase-binding protein (CD26) in goldfish brain. AB - The expression patterns of adenosine A(1) receptors (A(1)Rs), adenosine deaminase (ADA) and ADA binding protein (CD26) were studied in goldfish brain using mammalian monoclonal antibody against A(1)R and polyclonal antibodies against ADA and CD26. Western blot analysis revealed the presence of a band of 35 kDa for A(1)R in membrane preparations and a band of 43 kDa for ADA in both cytosol and membranes. Immunohistochemistry on goldfish brain slices showed that A(1) receptors were present in several neuronal cell bodies diffused in the telencephalon, cerebellum, optic tectum. In the rhombencephalon, large and medium sized neurons of the raphe nucleus showed a strong immunopositivity. A(1)R immunoreactivity was also present in the glial cells of the rhombencephalon and optic tectum. An analogous distribution was observed for ADA immunoreactivity. Tests for the presence of CD26 gave positive labelling in several populations of neurons in the rhombencephalon as well as in the radial glia of optic tectum, where immunostaining for ADA and A(1)R was observed. In goldfish astrocyte cultures the immunohistochemical staining of A(1)R, ADA and CD26, performed on the same cell population, displayed a complete overlapping distribution of the three antibodies. The parallel immunopositivity, at least in some discrete neuronal areas, for A(1)Rs, ADA and CD26 led us to hypothesize that a co localization among A(1)R, ecto-ADA and CD26 also exists in the neurons of goldfish since it has been established to exist in the neurons of mammals. Moreover, we have demonstrated for the first time, that A(1)R, ecto-ADA and CD26 co-localization is present on the astroglial component of the goldfish brain. This raises the possibility that a similar situation is also shown in the glia of the mammalian brain. PMID- 12547643 TI - Adenosine kinase and 5'-nucleotidase activity after prolonged wakefulness in the cortex and the basal forebrain of rat. AB - The effect of prolonged wakefulness on adenosine kinase (AK), ecto-5' nucleotidase and endo-5'-nucleotidase activity was assessed in the present study. Rats were sleep deprived for 3 or 6h, and one group was allowed to sleep 2h of recovery sleep after the 6h deprivation. The cortex and the basal forebrain were dissected, and frozen rapidly on dry ice. The enzyme activity of adenosine kinase was measured by monitoring the conversion of [2-3H]-adenosine into [3H]-adenosine monophosphate (AMP) and the ecto-5'-nucleotidase and endo-5'-nucleotidase activities by monitoring the conversion of [2-3H]-AMP into [3H]-adenosine. The enzyme activities did not change during deprivation or recovery sleep in either cortex or basal forebrain when compared to unhandled controls. Significant diurnal variation in enzyme activities was noted in both brain areas. In the basal forebrain adenosine kinase and both nucleotidases showed their lowest activity in the middle of the rest phase, 6h after lights on, suggesting a low level of adenosine metabolism, both production and degradation at this time point. In the cortex adenosine kinase had a diurnal activity pattern similar to the basal forebrain and the ecto-5'-nucleotidase activity was low already early in the rest phase, 3h after lights on, and remained low until the end part of the rest phase, 8h after lights on. Endo-5'-nucleotidase lacked diurnal variation. These activity patterns may be associated with the lower level of energy metabolism during sleep compared to wakefulness. PMID- 12547645 TI - Hyperoxia caused by microdialysis perfusion decreased striatal monoamines: involvement of oxidative stress. AB - Due to complex influence, such as utilization and permeability of arterial vessels to oxygen, there is a considerable difference of oxygen tension between extracellular fluid and perfusate usually used in microdialysis (30-60 Torr versus 145 Torr). Dialysate dopamine and monoamine metabolites-3,4 dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindoleacetic acid were measured under different kinds of oxygen tension solutions (145, 72, 48 Torr). In the acute and anesthetized group, dopamine, 3,4-dihydroxyphenylacetic acid, 5 hydroxyindoleacetic acid and homovanillic acid increased 72, 93, 86 and 65%, respectively when changing the perfusate from 145 Torr to near physiological 48 Torr, while in chronic and conscious group, carried out 72 h after surgery, these compounds showed obscure increases (only homovanillic acid produced a significant change of 14%). The different effect of perfusate oxygen tension on dialysate levels of monoamines in anesthetized and conscious rats might be caused by oxidative stress triggered by hyperoxia combined with anesthesia and surgical trauma. PMID- 12547646 TI - Neurotrophic and neurotoxic effects of zinc on neonatal cortical neurons. AB - Although zinc exerts direct neurotoxic action, this metal is also essential for the activity of numerous biological systems and zinc deficiency has been associated with various pathologies. We investigated the cellular responses and neuronal viability following exposure to different concentrations of zinc in primary cultures of neonatal rat cortical neurons. Higher concentrations of zinc (0.15 and 0.2 mM) triggered excessive zinc influx, glutathione depletion and ATP loss leading to necrotic neuronal death. In contrast, lower concentrations of zinc (0.05 and 0.1 mM) attenuated serum-deprivation induced apoptotic neuronal death. The antiapoptotic action of low amounts of zinc was found both in mixed cultures and neuron-enriched cultures indicating the independence of glial mediator. Neurotrophic action was not accompanied by significant alteration in those cellular responses but required chelatable zinc. The N-methyl-D-aspartate (NMDA) antagonist, MK-801, mimicked the beneficial effect of zinc in protecting neuronal death. Moreover, both MK-801 and zinc eliminated NMDA-induced neuronal injury. The results suggest that zinc is an intrinsic factor for neuron survival and exogenous zinc, in low amounts, is an active neuroprotectant against serum deprivation in part through the antagonism of NMDA receptor activation. PMID- 12547647 TI - Cloning and characterization of rat neuronal apoptosis inhibitory protein cDNA. AB - The human neuronal apoptosis inhibitory protein (NAIP) gene was originally discovered because of its deletion in infantile spinal muscular atrophy (SMA), a childhood genetic disorder characterized by motor neuron loss and progressive paralysis with muscular atrophy. Although SMA is now known to be caused by deletions of survival motor neuron (SMN), the fact that NAIP is an anti-apoptotic protein is consistent with the NAIP gene modifying SMA severity. Here we report the cloning of a 1.5 kb rat NAIP cDNA fragment which contains BIR-3 (third baculovirus inhibitory repeat) domain. This fragment shows 78% homology to the human NAIP and 86% homology to the murine counterpart. We have investigated the distribution of NAIP mRNA expressing neurons by in situ RT-PCR technique in the rat central nervous system (CNS). Although all of the neurons appeared to express NAIP mRNA ubiquitously, pronounced elevation of NAIP mRNA expression was observed in the areas innervated by glutamatergic neurons after kainic acid (KA) injection. We have raised an anti-rat NAIP antiserum in rabbits using NAIP cDNA and recombinant rat NAIP, and carried out an immunohistological investigation. We observed highly immunoreactive neuronal subpopulations in the retinal ganglion, cerebral cortex, hippocampus, basal forebrain, thalamus, areas of midbrain, Purkinje cells of the cerebellum, and motor neurons in the spinal cord. Increased immunoreactivity of glutamatergic neurons was also observed broadly in the CNS after KA treatment. This study provides additional evidence that expression of mRNA and gene products of NAIP seem to be regulated in response to excessive stimuli or injuries in rat CNS, and these results are compatible with an anti apoptotic role of NAIP in acute SMA as well as in brain injuries. PMID- 12547648 TI - Increase in secretion of glial cell line-derived neurotrophic factor from glial cell lines by inhibitors of vacuolar ATPase. AB - Glial cell line-derived neurotrophic factor (GDNF) was reported to be effective for treating subjects with neurodegenerative diseases such as Parkinson's disease. In search of finding a compound which promotes GDNF secretion, we found that concanamycin A (ConA), a vacuolar ATPase (V-type ATPase) inhibitor purified from Streptomyces diastatochromogens, enhanced GDNF secretion from glioma cells. The rat glioma cell line, C6, and the human glioma cell lines, U87MG and T98G, abundantly expressed GDNF mRNA, and secreted GDNF into culture media, and this event was potently enhanced by a Ca(2+) ionophore and by phorbol ester, as noted in other cells. ConA concentration dependently and potently increased GDNF release from C6, U87MG and T98G cells into culture media. In addition, ConA enhanced GDNF secretion from astrocyte primary cultures prepared from the human fetus with the same potency seen in glioma cell lines. Likewise, another V-type ATPase inhibitor, bafilomycinA1 facilitated GDNF release from C6, U87MG and T98G glioma cells, in a concentration-dependent manner. The potencies of these V-type ATPase inhibitors in enhancing GDNF secretion were consistent with those which inhibited V-type ATPase activity. These results suggest that blockade of V-type ATPase potently stimulates the secretion of GDNF from glial cells. The V-type ATPase inhibitors may be beneficial to use for the treatment of diseases in which increase in GDNF could be effective. PMID- 12547649 TI - Inherent desensitisation-preventing properties of a novel, subtype-selective AMPA receptor agonist, (S)-CPW 399, as a possible explanation for its excitotoxic action in cultured cerebellar granule cells. AB - The synthesis and pharmacological characterisation of (S)-CPW 399 as a novel, potent and subtype-selective agonist of the AMPA receptor was recently reported. Studies have been extended to investigate its excitotoxic action in primary cultures of mouse cerebellar granule cells. (S)-CPW 399 induced neuronal cell death in a time- and concentration-dependent manner (EC(50) approximately 70 microM) at 24-h exposure. (S)-CPW-induced neuronal death could be prevented by co administration with either of the AMPA/kainate selective receptor antagonists 6 cyano-7-nitro-quinoxaline-2,3-dione (CNQX) and 6-nitro-7 sulphamoylbenzo[f]quinoxaline-2,3-dione (NBQX) or by the 2,3-benzodiazepine, GYKI 53655 (a selective AMPA receptor antagonist); while no protection was afforded by either the NMDA receptor antagonist D,L(+/-)-2-amino-5-phosphonopentanoate (APV) or by nifedipine (an L-type calcium channel antagonist) when used alone or in combination. Cyclothiazide, which blocks AMPA receptor desensitisation, caused minimal potentiation of (S)-CPW 399-induced neuronal death, supporting accumulating evidence that (S)-CPW 399 is a full AMPA receptor agonist that markedly prevents a receptor desensitised conformation. (S)-AMPA, (S)-willardiine (a naturally-occurring heterocyclic excitatory amino acid) and its halogenated derivative, (S)-5-fluorowillardiine, had no deleterious effect on neuronal viability when used alone but each, in the presence of cyclothiazide, induced a concentration-dependent excitotoxic cell death with a rank order of potency (fluorowillardiine>>AMPA=willardiine). (S)-CPW 399 stimulated an increase in intracellular free-calcium levels ([Ca(2+)](i)) in a concentration-dependent fashion (EC(50) approximately 5 microM) attaining a value of six-fold that of 'resting' cells at maximum stimulation; achieved at approximately 100 microM (S) CPW 399. The (S)-CPW 399-stimulated increase in [Ca(2+)](i) was virtually abolished by GYKI 53655, NBQX, CNQX and by cobalt ions; markedly inhibited by nifedipine and marginally affected by D-APV. These results suggest that (S)-CPW 399 may be used as a pharmacological tool to aid in the investigation of the role of AMPA receptors in excitotoxicity and their molecular mechanisms of desensitisation. PMID- 12547650 TI - [3H]-ketanserin binding sites in different psychiatric disorders. AB - The results of the present study showed the presence of a high-affinity and saturable binding of [3H]-ketanserin to frontal and parietal brain membranes obtained postmortem from bipolar, depressed, schizophrenic patients and normal controls. The human brain samples (60 frontal cortex and 51 parietal cortex), were donated by the Stanley Foundation Brain Collection. The overall data showed that normal controls, depressed and schizophrenic patients had a higher density in the frontal than in the parietal cortex, while bipolar patients did not show any difference. When the data were analysed according to the two hemispheres, some additional, intriguing observations were made: it emerged that [3H] ketanserin binding sites did not show any difference in the two frontal cortices, while they were less represented in the right parietal cortex of normal and bipolar patients and more dense in schizophrenic patients. In conclusion, our study has demonstrated the presence of heterogenous alterations of [3H] ketanserin binding sites in healthy controls and different psychiatric disorders that may be of help in a further elucidation of the specific role that 5-HT(2A) receptors may play in these disorders. PMID- 12547651 TI - Cytochrome c translocation does not lead to caspase activation in maitotoxin treated SH-SY5Y neuroblastoma cells. AB - Cytosolic cytochrome c elevation has been associated with activation of caspase-3 like proteases. In this study, we demonstrate that treatment with the neurotoxin and potent calcium channel opener maitotoxin (MTX) induces cytochrome c release from the mitochondria that is not accompanied by caspase activation. Cytochrome c translocation in MTX-treated SH-SY5Y cells was readily apparent after 30 min and peaked at 2.5h. We assayed caspase activity by acetyl-Asp-Glu-Val-Asp-7-amido-4 methylcoumarin (Ac-DEVD-AMC) hydrolysis and by immunoblotting for caspase-3 processing and proteolysis of alphaII-spectrin and PARP. In contrast, treatment with pro-apoptosis agent staurosporine (STS) induced both cytochrome c release and caspase-3 activation after 2h. In addition, with MTX treatment, we found no evidence of caspase activation at any time point or MTX concentration used. Instead, we observed that caspase-9, Apaf-1 and caspase-3 were all partially truncated by calpain under these conditions. These combined effects likely contribute to the lack of caspase activation cascade in MTX-treated cells, despite the presence of cytochrome c in the cytosol. PMID- 12547652 TI - Effects of HIV protease inhibitor therapy on lipid metabolism. AB - Highly active antiretroviral therapy, which includes a combination of protease inhibitors, is highly successful in controlling human immunodeficiency virus (HIV) infection and reducing the morbidity and mortality of autoimmune deficiency syndrome (AIDS). However, the benefits of HIV protease inhibitors are compromised by numerous undesirable side effects. These include peripheral fat wasting and excessive central fat deposition (lipodystrophy), overt hyperlipidemia, and insulin resistance. The mechanism associated with protease inhibitor-induced metabolic abnormalities is multifactorial. One major effect of the protease inhibitor is its suppression of the breakdown of the nuclear form of sterol regulatory element binding proteins (nSREBP) in the liver and adipose tissues. Hepatic accumulation of nSREBP results in increased fatty acid and cholesterol biosynthesis, whereas nSREBP accumulation in adipose tissue causes lipodystrophy, reduces leptin expression, and promotes insulin resistance. The HIV protease inhibitors also suppress proteasome-mediated breakdown of nascent apolipoprotein (apo) B, thus resulting in the overproduction and secretion of triglyceride-rich lipoproteins. Finally, protease inhibitor also suppresses the inhibition of the glucose transporter GLUT-4 activity in adipose and muscle. This latter effect also contributes directly to insulin resistance and diabetes. These adverse effects need to be alleviated for long-term use of protease inhibitor therapy in treatment of HIV infection. PMID- 12547654 TI - Biosynthesis of phosphatidylcholine in bacteria. AB - Phosphatidylcholine (PC) is the major membrane-forming phospholipid in eukaryotes and can be synthesized by either of two pathways, the methylation pathway or the CDP-choline pathway. Many prokaryotes lack PC, but it can be found in significant amounts in membranes of rather diverse bacteria and based on genomic data, we estimate that more than 10% of all bacteria possess PC. Enzymatic methylation of phosphatidylethanolamine via the methylation pathway was thought to be the only biosynthetic pathway to yield PC in bacteria. However, a choline-dependent pathway for PC biosynthesis has been discovered in Sinorhizobium meliloti. In this pathway, PC synthase, condenses choline directly with CDP-diacylglyceride to form PC in one step. A number of symbiotic (Rhizobium leguminosarum, Mesorhizobium loti) and pathogenic (Agrobacterium tumefaciens, Brucella melitensis, Pseudomonas aeruginosa, Borrelia burgdorferi and Legionella pneumophila) bacteria seem to possess the PC synthase pathway and we suggest that the respective eukaryotic host functions as the provider of choline for this pathway. Pathogens entering their hosts through epithelia (Streptococcus pneumoniae, Haemophilus influenzae) require phosphocholine substitutions on their cell surface components that are biosynthetically also derived from choline supplied by the host. However, the incorporation of choline in these latter cases proceeds via choline phosphate and CDP-choline as intermediates. The occurrence of two intermediates in prokaryotes usually found as intermediates in the eukaryotic CDP-choline pathway for PC biosynthesis raises the question whether some bacteria might form PC via a CDP-choline pathway. PMID- 12547653 TI - Plasma platelet activating factor-acetylhydrolase (PAF-AH). AB - The platelet-activating factor-acetylhydrolase (PAF-AH) is an enzyme which catalyzes the hydrolysis of acetyl ester at the sn-2 position of PAF. The family of PAF-AHs consists of two intracellular isoforms (Ib and II), and one secreted isoform (plasma). These PAF-AHs show different biochemical characteristics and molecular structures. Plasma PAF-AH and intracellular isoform, II degrade not only PAF but also oxidatively fragmented phospholipids with potent biological activities. Among these PAF-AHs, plasma PAF-AH has been the target of many clinical studies in inflammatory diseases, such as asthma, sepsis, and vascular diseases, because the plasma PAF-AH activity in the patients with these diseases is altered when compared with normal individuals. Finding a genetic deficiency in the plasma PAF-AH opened the gate in elucidating the protecting role of this enzyme in inflammatory diseases. The most common loss-of-function mutation, V279F, is found in more than 30% of Japanese subjects (4% homozygous, 27% heterozygous). This single nucleotide polymorphism in plasma PAF-AH and the resulting enzymatic deficiency is thought to be a genetic risk factor in various inflammatory diseases in Japanese subjects. Administration of recombinant plasma PAF-AH or transfer of the plasma PAF-AH gene improves pathology in animal models. Therefore, substitution of plasma PAF-AH would be an effective in the treatment of the patients with the inflammatory diseases and a novel clinical approach. In addition, the detection of polymorphisms in the plasma PAF-AH gene and abnormalities in enzyme activity would be beneficial in the diagnosis of the inflammatory diseases. PMID- 12547656 TI - Pediatric and adolescent gynecology--the UK experience. AB - This review looks at the history of the establishment of pediatric and adolescent gynecology in the United Kingdom and, having addressed the chronology, sets the service in the context of the current health care system. The National Health Service does not provide an environment that encourages the delivery of this care in a structured way. There are some specialist services that are provided by central government, one of which addresses one part of pediatric and adolescent gynecology, but currently the services are relatively haphazard and delivered by individuals who have acquired knowledge and skills over time rather than in a structured training way. The need for improved services for adolescents and the training of doctors to provide this service is emphasized. PMID- 12547657 TI - Protruding giant cervical polyp in a young adolescent with a previous rhabdomyosarcoma. AB - Giant cervical polyps are rare entities, with only 8 reported cases in the international literature (MEDLINE 1966-2002). Of these cases, only one has been described in the pediatric population. There has been no report of a giant cervical polyp occurring in the older, although early, adolescent period. The discovery of a giant cervical polyp in a young adolescent is a gynecological oddity. The immense size of giant cervical polyps, together with their menacing appearance and unusual presentation, can simulate a malignant neoplasm. Care must be taken in their diagnosis and management so that the patient's fertility is not compromised. We report on a young adolescent girl with a previous history of a rhabdomyosarcoma (RMS), who presented with a giant cervical polyp protruding from the vaginal introitus. The literature on this entity is reviewed. PMID- 12547658 TI - Physical activity levels among urban adolescent females. AB - STUDY OBJECTIVE: To examine the proportion of girls engaging in vigorous physical activity in a sample of inner-city adolescent girls and to identify factors associated with this level of exercise. In addition, to report perceived barriers to exercise. DESIGN: Prospective cross-sectional survey. SETTING: Primary care health center. PARTICIPANTS: Females residing in a large northeastern city aged 12-21 yr (n = 305) who presented to a primary care site between September 2000 and May 2001 were consecutively recruited. MAIN OUTCOME MEASURE: In addition to being measured for height and weight, each subject anonymously completed a self report instrument assessing demographic and reproductive characteristics, weight concerns, number of friends who exercise, medical conditions limiting exercise, and efforts to alter weight status. To compare with other published research, we used standardized measures of vigorous physical activity. We also inquired about parent's weight, exercise, and health status. Finally, each subject was asked about access to physical education classes as well as perceived barriers to exercise. RESULTS: We found that 30.5% (n = 93) of inner-city females reported engaging in regular vigorous exercise in the last week, while 46.6% (n = 142) reported no physical activity. Five factors were significantly associated with regular vigorous exercise including: most or all friends exercised (AOR = 4.72); involved with a sports team (AOR = 3.59); trying to lose weight (AOR = 2.92); believing in the importance of exercise (AOR = 2.37; and being less than 17 years of age (AOR = 2.18). Time constraints and laziness were the most common reasons given for not engaging in physical activity. CONCLUSIONS: Among this sample of primarily minority, inner-city females, engaging in regular vigorous physical activity was much less than has been suggested by other studies. Research efforts to accurately measure physical activity levels among inner-city youth must continue concurrent to the development of health promotion interventions. PMID- 12547659 TI - Qualitative analysis of African-American adolescent females' beliefs about emergency contraceptive pills. AB - STUDY OBJECTIVE: This project was initiated to better understand why some African American adolescent females do not use emergency contraceptive pills (ECP), despite their widespread availability. Adolescents are considered likely candidates for ECP because they are more likely to inconsistently use birth control and have sporadic sexual behavior patterns. These factors increase the risk of unprotected intercourse and unplanned pregnancy. DESIGN: A qualitative study design was employed to assess the knowledge of African-American adolescent females regarding ECP and their beliefs affecting use and nonuse of ECP. SETTING: An adolescent clinic at a large, urban, academic children's hospital. PARTICIPANTS: Twenty-nine African-American adolescent (age 13-18) females.Interventions. Semi-structured, qualitative interviews lasting 15-20 minutes. MAIN OUTCOME MEASURES: Interviews yielded qualitative data that was coded and categorized into themes. RESULTS: Through the interviews, eight themes emerged: Taking Care of Self, Lack of Knowledge of ECP, Inaccurate Information, Sources of Information, Acceptability of Adolescent Pregnancy, Partner's Influence on Adolescent Choices, Discomfort Talking About Sex, and Concern About Sexually Transmitted Infections (STIs). CONCLUSIONS: The results from this qualitative study will contribute to future quantitative research efforts by providing insight into the decision-making process regarding ECP among this population. The themes provide a foundation for clinical implications and educational interventions. Although a large proportion of the sample (24%) had used ECP, misinformation about ECP and basic reproductive health issues was prevalent. Most of the adolescents report they obtain reproductive health information from a complex network of information sharing, in which their primary sources tend to be peers. PMID- 12547660 TI - Psychosocial predictors of pregnancy among low-income African-American adolescent females: a prospective analysis. AB - OBJECTIVE: To prospectively identify psychosocial predictors of pregnancy among African-American adolescent females. METHODS: African-American females, 14-18 yrs old, were recruited from schools and health clinics in low-income neighborhoods. Adolescents completed an in-depth survey and provided urine specimens for pregnancy testing at baseline and 6-month intervals for 1.5 years. Selected problem behaviors, demographic, and psychosocial variables were tested for bivariate and multivariate significance relative to biologically confirmed pregnancy during the follow-up period. Only adolescents who initially tested negative for pregnancy were included (n = 241). RESULTS: About 26% (n = 63) of the adolescents became pregnant over the follow-up period. Although a broad spectrum of variables achieved bivariate significance, few retained significance in the multivariate model. Multivariate predictors of pregnancy were biologically confirmed marijuana use (adjusted odds ratios [AOR] = 12.4, P = 0.0003) and perceiving that the sex partner desired pregnancy (AOR = 1.27, P = 0.01). A protective effect was observed for adolescents who reported that a family member received welfare benefits; these adolescents were about 60% less likely to become pregnant (AOR = 0.38, P = 0.04). CONCLUSIONS: Pediatricians and other health professionals who participate in community efforts to prevent first and subsequent adolescent pregnancies may benefit from recognizing that marijuana use and pregnancy may be co-occurring problems. Adolescents' perceptions of their boyfriends' level of desire for conception may also be an important predictor of pregnancy risk. The findings also suggest a possible protective effect of receiving TANF (Temporary Assistance to Needy Families) benefits; adolescent recipients of these programs may be more vigilant in their pregnancy prevention practices than those who are not recipients. PMID- 12547661 TI - Adolescents' attitudes and experiences regarding levonorgestrel 100 mcg/ethinyl estradiol 20 mcg. AB - STUDY OBJECTIVE: Oral contraceptive pills (OCs) are a commonly prescribed method of birth control for sexually experienced adolescents. The purpose of the current study was to describe anticipated parental involvement and adolescent compliance with pill taking, and to describe experiences with a 20-mcg OC regarding satisfaction and side effects after six cycles. DESIGN: Anticipated parental involvement and anticipated compliance, and experiences with an OC, were assessed at baseline, and after the first and sixth cycles during a clinical trial of oral contraceptives, which required parental consent. SETTING: Three adolescent medicine clinics in New York, NY; Cincinnati, OH; and San Juan, Puerto Rico. PARTICIPANTS: The sample consisted of 43 female adolescents with a mean age of 17 years. RESULTS: The majority of adolescents living with a parent anticipated parental involvement, and few adolescents anticipated difficulties with consistent pill taking. For two side effects (weight and mood changes), more than 30% of the adolescents anticipated the occurrence or worsening of side effects. However, few adolescents actually experienced increases in any of the nine side effects assessed. Ninety-seven percent of adolescents in this trial reported being satisfied with a 20-mcg OC. CONCLUSIONS: Health care providers can assess adolescents' anticipated difficulties with compliance with daily pill taking, and desire for help from their parents. This information can be incorporated into counseling to promote consistent and correct use. PMID- 12547662 TI - Self-collected vaginal swabs for the detection of multiple sexually transmitted infections in adolescent girls. AB - OBJECTIVE: To evaluate the use of self-collected vaginal swabs to test for Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas vaginalis, and to describe the acceptability of this technique to adolescents. DESIGN: Comparison of a new protocol for sexually transmitted infection (STI) testing with the current standard of care, using the same subjects. Survey of attitudes regarding the self-collection technique. SETTING: A juvenile correctional facility in western Pennsylvania. PARTICIPANTS: Convenience sample of 133 detainees, 12-17 years of age. INTERVENTION: Girls were invited to undergo STI screening using a self-collected vaginal swab. Polymerase chain reaction was used to test this specimen for each of the above three infections. Pelvic exams and endocervical testing were performed at the discretion of the physician performing the intake physical, independent of the study. MAIN OUTCOME MEASURES: The number of infections diagnosed using the self-testing protocol, compared to the number diagnosed using standard practices; acceptability of the self-collection technique. RESULTS: Twenty-four percent of sexually active subjects had one or more infections diagnosed by self-testing: 11.3% had C. trachomatis, 8.5% had N. gonorrhoeae, and 11.7% had T. vaginalis. Only 30% of subjects with infections had pelvic exams while detained; therefore 70% of girls with infections would have been missed in the absence of the self-testing option. The self-collection technique was acceptable to 95% of subjects. CONCLUSIONS: STI testing using self collected vaginal specimens is highly acceptable to adolescent girls, and can dramatically increase the detection rate for these three treatable infections when pelvic exams are not performed. PMID- 12547663 TI - Precocious puberty in a 3-year-old: too much, too little, too soon. PMID- 12547665 TI - Let's hear it for the Zena Princess Warriors. PMID- 12547664 TI - Hirsutism and adnexal masses in a teenager. PMID- 12547666 TI - Extended cycling of oral contraceptive pills for adolescents. PMID- 12547667 TI - Child and adolescent prostitution. PMID- 12547670 TI - Contextualizing the Genome: The Role of Epigenetics in Genetics, Development, and Evolution. November 25-28, 2001. Ghent, Belgium. Conference proceedings. PMID- 12547671 TI - Epigenetics: a challenge for genetics, evolution, and development? AB - In this paper, it is argued that differences in how one relates the genome to its surrounding contexts leads to diverse interpretations of the term epigenetics. Three different approaches are considered, ranging from gene-centrism, over gene regulation, to dynamic systems approaches. Although epigenetics receives its widest interpretation in a systems approach, a paradigmatic shift has taken place in biology from the abandonment of a gene-centric position on to the present. The epistemological and ontological consequences of this shift are made explicit. PMID- 12547672 TI - Theories in early embryology: close connections between epigenesis, preformationism, and self-organization. AB - In current biological and philosophical literature, the use of the terms epigenesis and epigenetics has increased tremendously. As these terms are often confused, this paper aims at clarifying the distinction between them by drawing their conceptual and historical evolutions. The evolution of the term epigenesis is situated in the context of early embryological studies. Departing from Aristotle's natural philosophy, it is shown that epigenesis gained alternating attention from the 17th century onwards, as it was introduced into neo-classical embryology and considered to be the opposite of the preformationist tradition. Where preformation stated that the germ cells of each organism contain preformed miniature adults that unfold during development, epigenesis held that the embryo forms by successive gradual exchanges in an amorphous zygote. Although both traditions tried to explain developmental organization, religious and metaphysical arguments on the conception of embryonic matter as either active or passive determined the scope of their respective explanations. It is shown that these very arguments still underlie the use of gene-centric metaphors in the molecular revolution of the 20th century. PMID- 12547673 TI - The relations between genetics and epigenetics: a historical point of view. AB - I have tried to unpack the polysemy of the word epigenetics by adopting a historical point of view and by focusing on the models that were proposed at the beginning of the 1960s to explain variations in gene activity during cell differentiation and development. Most of the questions that were or are at the core of epigenetics were posed in this period. This was due to the fact that the regulatory models and their extension to the notion of the genetic program were proposed as genetic answers to the questions raised by Waddington when he defined epigenetics in the 1940s. Studies of DNA methylation and chromatin structure, which became increasingly important in the 1960s and 1970s, were seen as alternative explanations to the regulatory mechanisms that had been previously proposed. This historical detour shows that epigenetics cannot be defined per se, but only as an evolving opposition to the piecemeal, reductionist approach of genetics. PMID- 12547674 TI - From epigenesis to epigenetics: the case of C. H. Waddington. AB - One continuous thread in this volume is the name of Conrad H. Waddington (1905 1975), the developmental biologist known as the inventor of the term epigenetics. After some biographical notes on his life, this article explores the meaning of the Waddingtonian equation and the context wherein it was developed. This equation holds that epigenesis + genetics = epigenetics, and refers in retrospect to the debate on epigenesis versus preformationism in neoclassical embryology. Whereas Waddington actualized this debate by linking epigenesis to developmental biology and preformation to genetics, thereby stressing the importance of genetic action in causal embryology, today's epigenetics more and more offers the possibility to enfeeble biological thinking in terms of genes only, as it expands the gene-centric view in biology by introducing a flexible and pragmatically oriented hierarchy of crucial genomic contexts that go beyond the organism. PMID- 12547675 TI - The changing concept of epigenetics. AB - We discuss the changing use of epigenetics, a term coined by Conrad Waddington in the 1940s, and how the epigenetic approach to development differs from the genetic approach. Originally, epigenetics referred to the study of the way genes and their products bring the phenotype into being. Today, it is primarily concerned with the mechanisms through which cells become committed to a particular form or function and through which that functional or structural state is then transmitted in cell lineages. We argue that modern epigenetics is important not only because it has practical significance for medicine, agriculture, and species conservation, but also because it has implications for the way in which we should view heredity and evolution. In particular, recognizing that there are epigenetic inheritance systems through which non-DNA variations can be transmitted in cell and organismal lineages broadens the concept of heredity and challenges the widely accepted gene-centered neo Darwinian version of Darwinism. PMID- 12547676 TI - What is "epi" about epigenetics? AB - What counts as epigenetic depends on what counts as genetic. It is argued that Weismannism, the doctrine of genetic continuity and somatic discontinuity, is the basis for an overly inclusive concept of epigenetics as every inherited resource "beyond the genes." An alternative theoretical perspective, the "reproducer" concept, is introduced to facilitate analysis of multiple inheritance systems without labeling all nongenetic inheritance "epigenetic." PMID- 12547677 TI - Genome organization and reorganization in evolution: formatting for computation and function. AB - This volume deals with the role of epigenetics in life and evolution. The most dynamic forms of functional genome formatting involve DNA interacting with cellular complexes that do not alter sequence information. Such important epigenetic phenomena are the main subjects of other articles in this volume. This article focuses on the long-lived form of genome formatting that lies within the DNA sequence itself. I argue for a computational view of genome function as the long-term information storage organelle of each cell. Structural formatting consists of organizing various signals and coding sequences into computationally ready systems facilitating genome expression and genome transmission. The basic features of genome organization can be understood by examining the E. coli lac operon as a paradigmatic genomic system. Multiple systems are connected through distributed signals and repetitive DNA to form higher-order genome system architectures. Molecular discoveries about mechanisms of DNA restructuring show that cells possess the natural genetic engineering functions necessary for evolutionary change by rearranging genomic components and reorganizing system architectures. The concepts of cellular computation and decision-making, genome system architecture, and natural genetic engineering combine to provide a new way of framing evolutionary theories and understanding genome sequence information. PMID- 12547678 TI - Alternative epigenetic states understood in terms of specific regulatory structures. AB - Generally speaking, epigenetic states or epigenetic regulation refer to situations in which several states of gene expression may coexist in similar environmental conditions, despite the absence of significant changes in the genomic sequence. In one way or another, the mechanisms behind these phenomena involve vicious circles, so that each epigenetic state tends to sustain itself, even after the disappearance of the inductive signal involved in the selection of that particular state. These vicious circles constitute positive feedback circuits and are found at the core of many developmental regulatory systems. In this paper, we present a qualitative model for the regulatory network formed by maternal and gap gene cross-regulations. This network controls the initial anterior-posterior patterning during early Drosophila embryogenesis and encompasses several intertwined feedback circuits. On the basis of our model analysis, we derive interesting insights about how specific expression states of the gap genes are selected along the anterior-posterior axis, in particular in relation with the activity of one positive feedback circuit, namely that formed by giant and Kruppel cross-inhibitions. In addition, we are able to qualitatively simulate the patterns of gene expression in the wild-type, as well as to predict the phenotypes of various loss-of-function mutations at the maternal and gap genes, or cis-regulatory mutations at the gap genes, as well as the effects of ectopic expression of these genes. PMID- 12547679 TI - On the roles of repetitive DNA elements in the context of a unified genomic epigenetic system. AB - Repetitive DNA sequences comprise a substantial portion of most eukaryotic and some prokaryotic chromosomes. Despite nearly forty years of research, the functions of various sequence families as a whole and their monomer units remain largely unknown. The inability to map specific functional roles onto many repetitive DNA elements (REs), coupled with the taxon-specificity of sequence families, have led many to speculate that these genomic components are "selfish" replicators generating genomic "junk." The purpose of this paper is to critically examine the selfishness, evolutionary effects, and functionality of REs. First, a brief overview of the range of ideas pertaining to RE function is presented. Second, the argument is presented that the selfish DNA "hypothesis" is actually a narrative scheme, that it serves to protect neo-Darwinian assumptions from criticism, and that this story is untestable and therefore not a hypothesis. Third, attempts to synthesize the selfish DNA concept with complex systems models of the genome and RE functionality are critiqued. Fourth, the supposed connection between RE-induced mutations and macroevolutionary events are stated to be at variance with empirical evidence and theoretical considerations. Hypotheses that base phylogenetic transitions in repetitive sequence changes thus remain speculative. Fifth and finally, the case is made for viewing REs as integrally functional components of chromosomes, genomes, and cells. It is argued throughout that a new conceptual framework is needed for understanding the roles of repetitive DNA in genomic/epigenetic systems, and that neo-Darwinian "narratives" have been the primary obstacle to elucidating the effects of these enigmatic components of chromosomes. PMID- 12547680 TI - Developmental robustness. AB - Developmental robustness, the capacity to stay "on track" despite the myriad vicissitudes that inevitably plague a developing organism, is, I argue, a prerequisite for natural selection and key to our understanding of the evolution of developmental processes. But how is such robustness achieved? And how can we reconcile this property with the delicate precision that seems to characterize so many developmental mechanisms, with what Michael Behe calls "irreducible complexity"? By looking at context, I argue. Developmental mechanisms must be robust with respect to the kinds of insults they are most likely to face, but with respect to less likely vicissitudes, they can be fragile. More specifically, I examine the relative absence of reaction-diffusion mechanisms in development and suggest that such mechanisms, theoretically attractive though they may be, have been judged by evolution to be ill suited for providing protection against the kinds of vicissitudes developing organisms are most likely to face, and have been supplanted by more intricate mechanisms that are protected from insult by structural design. PMID- 12547681 TI - The genome in its ecological context: philosophical perspectives on interspecies epigenesis. AB - Epigenesis concerns the interactions through which the inherited potentials of the genome become actualized into an adult organism. In addition to epigenetic interactions occurring within the developing embryo, there are also critical epigenetic interactions occurring between the embryo and its environment. These interactions can determine the sex of the embryo, increase its fitness, or even be involved in the formation of particular organs. This essay will outline the history of environmental concerns in developmental biology and provide some reasons for the decline and resurgence of these ideas, and it will then focus on two areas that have recently gained much attention: predator-induced polyphenisms and developmental symbioses. Research in these two areas of interspecies cooperation in morphogenesis has profound implications for what we consider to be normal development and how we proceed to study it. Studies of predator-induced polyphenism have shown that soluble factors from predators can change the development of prey in specific ways. Prey has evolved mechanisms to sense compounds released from their predators and to use these chemical cues to change their development in ways that prevent predation. New techniques in molecular biology, especially polymerase chain reaction and microarray analysis, have shown that symbioses between embryos and bacteria are widespread and that animals may use bacterial cues to complete their development. PMID- 12547682 TI - From representational preformationism to the epigenesis of openness to the world? Reflections on a new vision of the organism. AB - The problem of how to reconcile the apparent "purposiveness" of the living organism with nonteleological, mechanist modes of explanation was given a certain form through most of the 20th century by a relatively decontextualized understanding of the gene as the heritable determinant of phenotypic traits. As instrumentally preformationist presuppositions about genes give way to the burgeoning elucidation of cell and molecular mechanisms of epigenesis, basic questions about the nature of complex living systems and their evolutionary origins once again come into consideration. Some suggestions are offered for a vision of the genetically recontextualized organism. PMID- 12547683 TI - Floral biology and pollination mechanisms in two Viola species--from nectar to pollen flowers? AB - The genus Viola is represented by four related species in Brazil belonging to section Leptidium, one of the most primitive sections in the genus. Floral biology and pollination by bees were studied in Viola cerasifolia and V. subdimidiata in high-altitude areas in south-eastern Brazil. Flowers are zygomorphic and spurred. The five stamens are arranged in a cuff around the ovary, and pollen is released by means of apical connective projections, which form a cone surrounding the base of the style. The connective projections of the inferior stamens are elongated and curved to form a hook-shaped structure. Nectar secreting tissue can occur in the basal connective appendages of the inferior stamens, which project into the spur. Flowers of V. subdimidiata secreted a mean volume of 0.14 micro l nectar over a 24-h period; approx. 40 % of flowers did not secrete any nectar. The main pollinators of these Viola species are female bees belonging to the genus Anthrenoides (Andrenidae), which search mainly for pollen. These bees seem to be oligolectic and obtain large amounts of pollen from Viola by vibrating the flowers or by moving the hook repeatedly back and forth. Males of Anthrenoides patrol Viola clusters and also feed on nectar, acting as secondary pollinators. The basic floral structure in the genus Viola fits that of 'nectar flowers'. The uncommon hook-shaped projections, scanty nectar production, and behaviour of pollinators suggest that V. cerasifolia and V. subdimidiata are shifting their reward for pollinators from nectar to pollen. Based on floral morphology, this shift may be widespread in Viola sect. Leptidium. PMID- 12547684 TI - Simulation of leaf area development based on dry matter partitioning and specific leaf area for cut chrysanthemum. AB - This work aims to predict time courses of leaf area index (LAI) based on dry matter partitioning into the leaves and on specific leaf area of newly formed leaf biomass (SLA(n)) for year-round cut chrysanthemum crops. In five glasshouse experiments, each consisting of several plant densities and planted throughout the year, periodic destructive measurements were conducted to develop empirical models for partitioning and for SLA(n). Dry matter partitioning into leaves, calculated as incremental leaf dry mass divided by incremental shoot dry mass between two destructive harvests, could be described accurately (R(2 )= 0.93) by a Gompertz function of relative time, R(t). R(t) is 0 at planting date, 1 at the start of short-days, and 2 at final harvest. SLA(n), calculated as the slope of a linear regression between periodic measurements of leaf dry mass (LDM) and LAI, showed a significant linear increase with the inverse of the daily incident photosynthetically active radiation (incident PAR, MJ m(-2 )d(-1)), averaged over the whole growing period, the average glasshouse temperature and plant density (R(2 )= 0.74). The models were validated by two independent experiments and with data from three commercial growers, each with four planting dates. Measured shoot dry mass increase, initial LAI and LDM, plant density, daily temperature and incident PAR were input into the model. Dynamics of LDM and LAI were predicted accurately by the model, although in the last part of the cultivation LAI was often overestimated. The slope of the linear regression of simulated against measured LDM varied between 0.95 and 1.09. For LAI this slope varied between 1.01 and 1.12. The models presented in this study are important for the development of a photosynthesis-driven crop growth model for cut chrysanthemum crops. PMID- 12547685 TI - Genetic diversity in Primula obconica (Primulaceae) from central and south-west China as revealed by ISSR markers. AB - Genetic diversity was investigated in 60 individuals of Primula obconica from four natural populations (from Hubei, central China, and Sichuan, south-west China) and from cultivated material. Inter-simple sequence repeat (ISSR) techniques produced 249 polymorphic bands and identified 60 ISSR genotypes. Based on Shannon's index and Nei's genetic diversity, the genetic diversity detected in all natural populations of P. obconica was much higher than that in the cultivated plants, and that in the three Dalaoling (Hubei) populations was higher than that in the Wawushan (Sichuan) population. UPGMA cluster analysis showed that there was no distinct genetic differentiation between populations from the Mt Dalaoling area and the Mt Wawushan area. This study provides a population level genetic profile of P. obconica, which was previously poorly known but which is important for Primula breeding and cultivation. PMID- 12547686 TI - Fungicide sprays can injure the stigmatic surface during receptivity in almond flowers. AB - Fungicides can be detrimental to flower development, pollen function and fruit set in a number of crops. Almond is a self-incompatible nut crop that has a fruit set of only approx. 30 % of the total number of flowers. Thus, interference of pollination and fertilization by fungicide sprays is of concern, and identification of chemicals having the least detrimental effects would be desirable. The objective of this study was to evaluate the effect of fungicide sprays on stigma morphology in almond using a laboratory spray apparatus that simulated field applications. Four fungicides (azoxystrobin, myclobutanil, iprodione and cyprodinil) were applied, and fresh, unfixed stigmatic surfaces were observed using a scanning electron microscope at 4 and 24 h after spraying. Increased exudate accumulation was induced by azoxystrobin at both time periods, and localized damage and collapse of stigmatic cells were observed after 24 h. Damaged stigmatic papillae exhibited wrinkling, surface distortion or collapse. Likewise, myclobutanil caused significant damage to and collapse of papillae; these were more extensive at later observations. Iprodione had no effect on exudate accumulation but caused marked and severe collapse of stigmatic papillae which was pronounced at 24 h. Cyprodinil promoted a copious increase in exudate secretion and caused the most severe collapse of stigmatic cells of all the fungicides evaluated. Damage was somewhat localized at 4 h but more global at 24 h. This study has verified that certain fungicide sprays have direct detrimental effects on stigma morphology and enhance exudate production in almond flowers. PMID- 12547687 TI - Low temperature enhances photosynthetic down-regulation in French bean (Phaseolus vulgaris L.) plants. AB - The mechanisms of photosynthetic adaptation to different combinations of temperature and irradiance during growth, and especially the consequences of exposure to high light (2000 micro mol m(-2) s(-1) PPFD) for 5 min, simulating natural sunflecks, was studied in bean plants (Phaseolus vulgaris L.). A protocol using only short (3 min) dark pre-treatment was introduced to maximize the amount of replication possible in studies of chlorophyll fluorescence. High light at low temperature (10 degrees C) significantly down-regulated photosynthetic electron transport capacity [as measured by the efficiency of photosystem II (PSII)], with the protective acclimation allowing the simulated sunflecks to be used more effectively for photosynthesis by plants grown in low light. The greater energy dissipation by thermal processes (lower F(v)'/F(m)' ratio) at low temperature was related to increased xanthophyll de-epoxidation and to the fact that photosynthetic carbon fixation was more limiting at low than at high temperatures. A key objective was to investigate the role of photorespiration in acclimation to irradiance and temperature by comparing the effect of normal (21 kPa) and low (1.5 kPa) O(2) concentrations. Low [O(2)] decreased F(v)/F(m) and the efficiency of PSII (Phi(PSII)), related to greater PSII down-regulation in cold pre-treated plants, but minimized further inhibition by the mild 'sunfleck' treatment used. Results support the hypothesis that photorespiration provides a 'safety-valve' for excess energy. PMID- 12547688 TI - Photosynthetic induction and leaf carbon gain in the tropical understorey epiphyte, Aspasia principissa. AB - Gas exchange of the understorey epiphyte Aspasia principissa was studied in fluctuating light conditions both in the laboratory and in the field, testing the hypothesis that vascular epiphytes differ from most terrestrial understorey plants in showing a higher priority for water conservation. Consequently, a slow response of stomatal conductance to sudden increases in incident photon flux density (PFD) was expected, as was a fast loss of induction after such a light fleck. Results were only partly consistent with these expectations. Full induction of photosynthesis was indeed very slow and was not reached before, respectively, 40 and 60 min of saturating PFD in the field and the laboratory. In contrast, kinetics of induction loss were comparable with those of most terrestrial species studied to date. The overall impact of light flecks on in situ carbon gain again fulfilled expectations, being rather limited: the observed carbon gain was only approx. 66% of the potential carbon gain estimated from a square-wave response model. It is concluded that in the drought-prone epiphytic habitat of a moist lowland forest, water conservation takes priority over carbon gain, which severely limits the use of light flecks for CO(2) fixation in vascular epiphytes. PMID- 12547689 TI - A flexible sigmoid function of determinate growth. AB - A new empirical equation for the sigmoid pattern of determinate growth, 'the beta growth function', is presented. It calculates weight (w) in dependence of time, using the following three parameters: t(m), the time at which the maximum growth rate is obtained; t(e), the time at the end of growth; and w(max), the maximal value for w, which is achieved at t(e). The beta growth function was compared with four classical (logistic, Richards, Gompertz and Weibull) growth equations, and two expolinear equations. All equations described successfully the sigmoid dynamics of seed filling, plant growth and crop biomass production. However, differences were found in estimating w(max). Features of the beta function are: (1) like the Richards equation it is flexible in describing various asymmetrical sigmoid patterns (its symmetrical form is a cubic polynomial); (2) like the logistic and the Gompertz equations its parameters are numerically stable in statistical estimation; (3) like the Weibull function it predicts zero mass at time zero, but its extension to deal with various initial conditions can be easily obtained; (4) relative to the truncated expolinear equation it provides more reasonable estimates of final quantity and duration of a growth process. In addition, the new function predicts a zero growth rate at both the start and end of a precisely defined growth period. Therefore, it is unique for dealing with determinate growth, and is more suitable than other functions for embedding in process-based crop simulation models to describe the dynamics of organs as sinks to absorb assimilates. Because its parameters correspond to growth traits of interest to crop scientists, the beta growth function is suitable for characterization of environmental and genotypic influences on growth processes. However, it is not suitable for estimating maximum relative growth rate to characterize early growth that is expected to be close to exponential. PMID- 12547690 TI - Response of potato tuber cell division and growth to shade and elevated CO2. AB - Plants adjust their sink-organ growth rates, development and distribution of dry matter in response to whole-plant photosynthate status. To advance understanding of these processes, potato (Solanum tuberosum L.) plants were subjected to CO(2) and light flux treatments, and early tuber growth was assessed. Atmospheric CO(2) (700 or 350 micro mol mol(-1)) and light flux (shade and control illumination) treatments were imposed at two growth stages: tuber initiation (TI) and tuber bulking (TB). Elevated CO(2) increased accumulation of total net biomass when imposed at both stages, and increased tuber growth rate by about 36 %, but did not increase the number of tubers. Elevated CO(2) increased the number of cells in tubers at both TI and TB stages, whereas shade substantially decreased the number of cells at both stages. Generally, treatments did not affect cell volume or the proportion of nuclei endoreduplicating (repeated nuclear DNA replication in the absence of cell division), but the shade treatment led to a decrease in cell volume at TB and a decrease in endoreduplication at TI. Elevated CO(2) increased, and shade decreased, glucose concentration and soluble invertase activity in the cambial zones at both TI and TB, whereas sucrose concentration and activities of glucokinase, fructokinase, cell-wall-bound invertase and thymidine kinase were unaffected. Modulation of tuber cell division was responsible for much of the growth response to whole-plant photosynthate status, and treatments affected cambial-zone glucose and soluble invertase in a pattern suggesting involvement of a glucose signalling pathway. PMID- 12547691 TI - Modelling cereal root systems for water and nitrogen capture: towards an economic optimum. AB - A quantitative model of wheat root systems is developed that links the size and distribution of the root system to the capture of water and nitrogen (which are assumed to be evenly distributed with depth) during grain filling, and allows estimates of the economic consequences of this capture to be assessed. A particular feature of the model is its use of summarizing concepts, and reliance on only the minimum number of parameters (each with a clear biological meaning). The model is then used to provide an economic sensitivity analysis of possible target characteristics for manipulating root systems. These characteristics were: root distribution with depth, proportional dry matter partitioning to roots, resource capture coefficients, shoot dry weight at anthesis, specific root weight and water use efficiency. From the current estimates of parameters it is concluded that a larger investment by the crop in fine roots at depth in the soil, and less proliferation of roots in surface layers, would improve yields by accessing extra resources. The economic return on investment in roots for water capture was twice that of the same amount invested for nitrogen capture. PMID- 12547692 TI - Variation in onset of summer dormancy and flowering capacity along an aridity gradient in Poa bulbosa L., a geophytic perennial grass. AB - Variation in the onset of summer dormancy and flowering capacity of 16 populations of Poa bulbosa, collected along a steep north-south aridity gradient in Israel (810-110 mm rain year(-1)), was studied under controlled conditions in a phytotron (16 h daylength, 22/16 degrees C day/night) and under natural conditions in a garden experiment in a net-house. Plant age at the onset of dormancy varied markedly amongst populations (7-16 weeks under controlled conditions) and was positively correlated with mean annual precipitation at the site of origin of the population, i.e. dormancy was earlier as aridity increased. Flowering capacity in the different populations was negatively correlated with rainfall in the original habitat and, consequently, also with the age at onset of dormancy, i.e. the lower the mean annual precipitation, the earlier the onset of dormancy and the higher the proportion of flowering plants and panicles per plant. Differences in xeromorphic leaf traits were also observed among populations from locations differing in aridity. Plants from the more arid sites (110-310 mm year(-1)) generally had greyish and curved leaves, whereas plants from more humid sites (500-810 mm year(-1)) tended to have green and straight leaves. Thus, plants with curved and/or greyish leaves generally had a higher flowering capacity and entered dormancy earlier than plants with straight and/or green leaves. The significance of the association among these traits for the adaptation of P. bulbosa to increasing aridity is discussed. PMID- 12547694 TI - Complement in ischemia reperfusion injury. PMID- 12547695 TI - Nailing down a link between tuberin and renal cysts. PMID- 12547696 TI - Multicolor deconvolution microscopy of thick biological specimens. AB - One limitation in understanding disease at the cellular level has been the inability to efficiently analyze DNA on a cell-to-cell basis within the natural tissue context. However, DNA analyses at a single-cell resolution should be instrumental for the understanding of cancer cell biology, cancer evolution, for chromosomal mosaic analysis and rare cell events, and should provide otherwise inaccessible information on essential biological processes. Here we present a fluorescence in situ hybridization-based multicolor deconvolution technique for three-dimensional microscopy. We use up to seven different color channels for probe detection, which allows the simultaneous high-resolution localization of multiple point-like sources within a biological specimen with a thickness of up to 30 micro m. In addition, a DNA counterstain is used for volume labeling of the nuclei offering the opportunity for a simultaneous segmentation of nuclei. Furthermore, as the instrumentation consists of a standard fluorescence microscope it represents a low-cost method as compared to confocal microscopy. PMID- 12547697 TI - In situ hybridization AT-tailing with catalyzed signal amplification for sensitive and specific in situ detection of human immunodeficiency virus-1 mRNA in formalin-fixed and paraffin-embedded tissues. AB - In situ hybridization is one of the most important techniques to visualize gene expression at the cellular level in various tissues. The in situ hybridization-AT tailing (ISH-AT) method uses a specially designed and synthesized oligonucleotide probe that has (AT)10 on the 3' side. This (AT)10 of the probe is elongated by DeltaTth DNA polymerase in the presence of dATP, dTTP, and labeled dUTP in the tissue after hybridization. Through this process the target is labeled with many hapten molecules. In this study, we detected human immunodeficiency virus type 1 RNA in formalin-fixed and paraffin-embedded tissues obtained from autopsied patients with acquired immunodeficiency syndrome by combining ISH-AT with the catalyzed signal amplification (CSA) system (ISH-AT-CSA), although we failed to detect signals from the same samples by conventional in situ hybridization using RNA probes (RISH) with CSA (RISH-CSA). We demonstrated that the ISH-AT-CSA method was superior to RISH-CSA in terms of both sensitivity and specificity, and that it was applicable to fluorescence in situ hybridization and double staining with immunohistochemistry for the characterization of cell phenotypes. PMID- 12547698 TI - Epithelial to mesenchymal transition in human breast cancer can provide a nonmalignant stroma. AB - A breast carcinoma biopsy showed cytochemical evidence of epithelial mesenchymal transition and an alpha-smooth muscle actin-positive stromal reaction. To study the lineage, and the nature of the cells in the stromal reaction, we derived a novel cell line, HBFL-1, from the explanted biopsy. HBFL-1 cells are immortal and exhibit a shared non-random X-chromosome inactivation pattern with the epithelial tumor of origin. Yet they closely resemble normal, finite-life-span fibroblasts by morphology, lack of tumor formation in nude mice, marker expression profile, protein pattern using two-dimensional gel electrophoresis and the ability to undergo myofibroblast conversion. HBFL-1 interacts reciprocally with tumor cells in collagen gel to induce activation of MMP2, leading to tumor-like behavior of epithelial colonies. In vivo, HBFL-1 cells resembled normal-derived myofibroblasts and conferred a significant 3.5- to 7-fold increase in MCF-7 tumor size in nude mice. However, that they were indeed not normal fibroblasts was revealed by residual keratin expression and formation of epithelial microfoci in a reconstituted basement membrane and in nude mice. We conclude that breast cancer can generate its own nonmalignant stroma and that one function for this is that of a reciprocal interaction with epithelial tumor cells to facilitate tumor growth. PMID- 12547699 TI - Arrest of B16 melanoma cells in the mouse pulmonary microcirculation induces endothelial nitric oxide synthase-dependent nitric oxide release that is cytotoxic to the tumor cells. AB - Metastatic cancer cells seed the lung via blood vessels. Because endothelial cells generate nitric oxide (NO) in response to shear stress, we postulated that the arrest of cancer cells in the pulmonary microcirculation causes the release of NO in the lung. After intravenous injection of B16F1 melanoma cells, pulmonary NO increased sevenfold throughout 20 minutes and approached basal levels by 4 hours. NO induction was blocked by N(G)-nitro-L-arginine methyl ester (L-NAME) and was not observed in endothelial nitric oxide synthase (eNOS)-deficient mice. NO production, visualized ex vivo with the fluorescent NO probe diaminofluorescein diacetate, increased rapidly at the site of tumor cell arrest, and continued to increase throughout 20 minutes. Arrested tumor cells underwent apoptosis with apoptotic counts more than threefold over baseline at 8 and 48 hours. Neither the NO signals nor increased apoptosis were seen in eNOS knockout mice or mice pretreated with L-NAME. At 48 hours, 83% of the arrested cells had cleared from the lungs of wild-type mice but only approximately 55% of the cells cleared from eNOS-deficient or L-NAME pretreated mice. eNOS knockout and L-NAME treated mice had twofold to fivefold more metastases than wild-type mice, measured by the number of surface nodules or by histomorphometry. We conclude that tumor cell arrest in the pulmonary microcirculation induces eNOS-dependent NO release by the endothelium adjacent to the arrested tumor cells and that NO is one factor that causes tumor cell apoptosis, clearance from the lung, and inhibition of metastasis. PMID- 12547700 TI - Apolipoprotein B in cholesterol-containing drusen and basal deposits of human eyes with age-related maculopathy. AB - Lipids accumulate in Bruch's membrane (BrM), a specialized vascular intima of the eye, and in extracellular lesions associated with aging and age-related maculopathy (ARM). We tested the hypothesis that ARM and atherosclerotic cardiovascular disease share molecules and mechanisms pertaining to extracellular lipid accumulation by localizing cholesterol and apolipoprotein B (apo B) in BrM, basal deposits, and drusen. Human donor eyes were preserved <4 hours postmortem and cryosectioned. Sections were stained with traditional lipid stains and filipin for esterified and unesterified cholesterol or probed with antibodies to apo B, apo E, and apo C-III. Normal adult retinal pigment epithelium (RPE) was subjected to RT-PCR and Western blot analysis for apolipoprotein mRNA and protein. Esterified and unesterified cholesterol was present in all drusen and basal deposits of ARM and normal eyes. Both apo B and apo E but not apo C-III were found in BrM, drusen, and basal deposits. Fewer macular drusen were stained by traditional lipid stains and apolipoprotein antibodies than peripheral drusen. RPE contained apo B and apo E mRNA and protein. Finding cholesterol and apo B in sub-RPE deposits links ARM with important molecules and mechanisms in atherosclerosis initiation and progression. The combination of apo B mRNA and protein in RPE raises the possibility that intraocular assembly of apo B containing lipoproteins is a pathway involved in forming cholesterol-enriched lesions in ARM. PMID- 12547701 TI - CC chemokine receptor 8 in the central nervous system is associated with phagocytic macrophages. AB - CC chemokine receptor 8 (CCR8) has been detected in vitro on type 2 helper and regulatory lymphocytes, which might exert beneficial functions in multiple sclerosis (MS) and on macrophages and microglia, possibly promoting tissue injury in MS lesions. To discriminate the relevant expression pattern in vivo, we defined the cell types that expressed CCR8 in MS lesions and determined the relationship of CCR8 expression and demyelinating activity. CCR8 was not expressed on T cells but was associated with phagocytic macrophages and activated microglia in MS lesions and directly correlated with demyelinating activity. To identify factors associated with CCR8 expression, the study was extended to other central nervous system (CNS) pathologies. CCR8 was consistently expressed on phagocytic macrophages and activated microglia in stroke and progressive multifocal leukoencephalopathy, but not expressed on microglia in pathologies that lacked phagocytic macrophages such as senile change of the Alzheimer's type. CCR8 was up-regulated by macrophage differentiation and activating stimuli in vitro. In summary CNS CCR8 expression was associated with phagocytic macrophages and activated microglial cells in human CNS diseases, suggesting that CCR8 may be a feasible target for therapeutic intervention in MS. CCR8 expression may also indicate a selective program of mononuclear phagocyte gene expression. PMID- 12547702 TI - Increased furin activity enhances the malignant phenotype of human head and neck cancer cells. AB - Many proteins are synthesized as inactive proforms requiring a proteolytic processing to render them active. A variety of proteases catalyze these cleavage reactions. Proprotein convertases are a family of serine proteases capable of activating substrates that will subsequently intervene in extracellular matrix (ECM) degradation, cell growth, differentiation and viral pathogenesis. Furin, the prototype of this family, has been implicated in many physiological and pathological processes. Some of its substrates such as TGF-beta, MT-MMP's, and IGFR-1 have been identified. Overexpression of furin has been observed in several human tumors. In this report we demonstrate that overexpression of furin causes a significant increase in the invasive potential of human tumor cells of low and moderate aggressive potential in vitro and in vivo. SCC12 and SCC15 were transfected with furin cDNA, resulting in efficient processing of furin substrates. An in vivo invasion assay showed enhancement of invasive ability. Inhibition of furin activity with the synthetic inhibitor decanoyl-Arg-Val-Lys Arg-chloromethyl-ketone, CMK, showed a significant decrease in both processing and in vitro invasiveness. A moderate enhancement in proliferation rate was observed when cells were transfected with furin. CMK treatment resulted in a marked reduction of this effect. Tumors obtained after subcutaneous (s.c.) inoculation of furin-overexpressing cells were larger and developed earlier than the controls. Furin overexpression caused an imbalance in the activation of invasion and proliferation-related substrates leading to the acquisition of an advanced malignant phenotype. In addition, inhibition of furin activity decreases substrate activation, proliferation rate, and invasive potential of cancer cells, suggesting that furin is a potentially useful target for therapeutics. PMID- 12547703 TI - Role for the alternative complement pathway in ischemia/reperfusion injury. AB - The terminal complement components play an important role in mediating tissue injury after ischemia and reperfusion (I/R) injury in rats and mice. However, the specific complement pathways involved in I/R injury are unknown. The role of the alternative pathway in I/R injury may be particularly important, as it amplifies complement activation and deposition. In this study, the role of the alternative pathway in I/R injury was evaluated using factor D-deficient (-/-) and heterozygote (+/-) mice. Gastrointestinal ischemia (GI) was induced by clamping the mesenteric artery for 20 minutes and then reperfused for 3 hours. Sham operated control mice (+/- versus -/-) had similar baseline intestinal lactate dehydrogenase activity (P = ns). Intestinal lactate dehydrogenase activity was greater in -/- mice compared to +/- mice after GI/R (P = 0.02) thus demonstrating protection in the -/- mice. Intestinal myeloperoxidase activity in +/- mice was significantly greater than -/- mice after GI/R (P < 0.001). Pulmonary myeloperoxidase activity after GI/R was significantly higher in +/- than -/- mice (P = 0.03). Addition of human factor D to -/- animals restored GI/R injury and was prevented by a functionally inhibitory antibody against human factor D. These data suggest that the alternative complement pathway plays an important role in local and remote tissue injury after GI/R. Inhibition of factor D may represent an effective therapeutic approach for GI/R injury. PMID- 12547704 TI - Polycystic kidney disease as a result of loss of the tuberous sclerosis 2 tumor suppressor gene during development. AB - Somatic loss of function of the tuberous sclerosis 2 (TSC2) tumor suppressor gene leads to the development of benign and malignant lesions of the kidney, brain, uterus, spleen, and liver and germline loss of function of this tumor suppressor gene is embryonic lethal. In addition, the gene product of TSC2, tuberin, is necessary for normal function of the polycystic kidney disease 1 (PKD1) gene product, polycystin-1, which is required for normal cell-cell and cell-matrix interactions. We report here the development of severe polycystic kidney disease in three cases of young Eker rats carrying a germline inactivation of one allele of the Tsc2 gene. Extrarenal tumors were also noted in the spleen and uterus of these animals, which was remarkable given their young age and in the case of the spleen, diffuse involvement of the affected organ. A cell line (EKT2) was established from an affected kidney of one of these animals and used in conjunction with tissues from affected animals to elucidate the defect responsible for the development of these lesions. Affected cells were determined to have lost the wild-type Tsc2 allele while retaining two copies of chromosome 10 containing the mutant Tsc2 allele along with two normal copies of the Pkd1 gene. The genetic data, bilateral nature of the observed kidney disease, and extent of involvement of the spleen and kidney indicate that, in affected animals, loss of the wild-type Tsc2 allele occurred during embryogenesis, probably as a result of chromosome nondisjunction, with affected animals being mosaics for loss of Tsc2 gene function. PMID- 12547705 TI - Conventional and tissue microarray immunohistochemical expression analysis of mismatch repair in hereditary colorectal tumors. AB - Immunohistochemistry (IHC) of mismatch repair (MMR) proteins in colorectal tumors together with microsatellite analysis (MSI) can be helpful in identifying families eligible for mutation analysis. The aims were to determine sensitivity of IHC for MLH1, MSH2, and MSH6 and MSI analysis in tumors from known MMR gene mutation carriers; and to evaluate the use of tissue microarrays for IHC (IHC TMA) of colon tumors in its ability to identify potential carriers of MMR gene mutations, and compare it with IHC on whole slides. IHC on whole slides was performed in colorectal tumors from 45 carriers of a germline mutation in one of the MMR genes. The TMA cohort consisted of 129 colon tumors from (suspected) hereditary nonpolyposis colorectal cancer (HNPCC) patients. Whole slide IHC analysis had a sensitivity of 89% in detecting MMR deficiency in carriers of a pathogenic MMR mutation. Sensitivity by MSI analysis was 93%. IHC can also be used to predict which gene is expected to harbor the mutation: for MLH1, MSH2, and MSH6, IHC on whole slides would have correctly predicted the mutation in 48%, 92%, and 75% of the cases, respectively. We propose a scheme for the diagnostic approach of families with (suspected) HNPCC. Comparison of the IHC results based on whole slides versus TMA, showed a concordance of 85%, 95%, and 75% for MLH, MSH2, and MSH6, respectively. This study therefore shows that IHC-TMA can be reliably used to simultaneously screen a large number of tumors from (suspected) HNPCC patients, at first in a research setting. PMID- 12547706 TI - Localization of the PP2A B56gamma regulatory subunit at the Golgi complex: possible role in vesicle transport and migration. AB - The BL6 subline was derived from the F10 line, which was derived from the B16 mouse melanoma cell line. BL6 cells are more invasive than F10 cells and differ genetically from F10 cells by an alteration of the gene encoding the B56gamma regulatory subunit of protein phosphatase 2A (PP2A). This alteration results in the transcription of mRNA encoding a truncated variant of the B56gamma1 isoform (Deltagamma1). When F10 cells were stained with a polyclonal antibody that recognizes three B56gamma isoforms, B56gamma1, B56gamma2, and B56gamma3, the immunofluorescent signals co-localized well with the cis-Golgi marker proteins. When BL6 cells were fractionated in a sucrose gradient, B56gamma1 and B56gamma2, but not B56gamma3, were present in the Golgi-enriched fraction. This fraction also contained the catalytic subunit of PP2A. FLAG-tagged Deltagamma1 preferentially localized to the trans-Golgi area rather than the cis-Golgi. This localization was the same as that of FLAG-tagged B56gamma1. NIH3T3 cells stably expressing Deltagamma1 transported a mutant viral protein from the endoplasmic reticulum to the plasma membrane much faster than wild-type cells. Their directional migration, as assessed by the advance of cells into a cell-free area, was also elevated. As Deltagamma1 reduces the activity of the B56gamma-containing PP2A holoenzymes, these results suggest that the normal holoenzymes suppress vesicle transport and that Deltagamma1 might increase the invasive ability of BL6 cells by activating Golgi function. PMID- 12547707 TI - Renal angiomyolipomas from patients with sporadic lymphangiomyomatosis contain both neoplastic and non-neoplastic vascular structures. AB - Renal angiomyolipomas are highly vascular tumors that occur sporadically, in women with pulmonary lymphangiomyomatosis (LAM), and in tuberous sclerosis complex (TSC). The goal of this study was to determine whether the distinctive vessels of angiomyolipomas are neoplastic or reactive. We studied angiomyolipomas with loss of heterozygosity (LOH) in the TSC2 region of chromosome 16p13 from patients with LAM. We found that angiomyolipomas contain five morphologically distinct vessel types: cellular, collagenous, hemangiopericytic, glomeruloid, and aneurysmatic. Using laser capture microdissection, we determined that four of the vessel types have TSC2 LOH and are therefore neoplastic. One vessel type, collagenous vessels, did not have LOH, and is presumably reactive. Recently, activation of S6 Kinase and its target S6 ribosomal protein (S6) was demonstrated in cells lacking TSC2 expression. We found that angiomyolipoma vessel types in which LOH were detected were immunoreactive with anti-phospho-S6 antibodies. Angiomyolipoma cells without LOH, including the endothelial component of the vessels, were not immunoreactive. To our knowledge, angiomyolipomas are the first benign vascular tumor in which the vascular cells, rather than the stromal cells, have been found to be neoplastic. Angiomyolipomas appear to reflect novel vascular mechanisms that may be the result of activation of cellular pathways involving S6 Kinase. PMID- 12547708 TI - Altered pattern of major histocompatibility complex expression in renal carcinoma: tumor-specific expression of the nonclassical human leukocyte antigen G molecule is restricted to clear cell carcinoma while up-regulation of other major histocompatibility complex antigens is primarily distributed in all subtypes of renal carcinoma. AB - Renal epithelial cancers represent a heterogeneous group of neoplasms arising from the malignant transformation of presumed diverse cell lineages. We recently demonstrated that tumor-specific up-regulation of human leukocyte antigen (HLA) G, a nonclassical HLA class Ib molecule that might be involved in immune evasion by tumor cells, frequently occurs in conventional (clear cell) renal carcinoma. We here examined whether HLA-G activation is a common process affecting all types of renal epithelial tumors. We analyzed a series of 38 paraffin-embedded tumors including clear cell, papillary, chromophobe, collecting duct carcinoma, and oncocytoma. Seven of 12 (58%) clear cell tumors were positive by immunohistochemistry, whereas all of the other subtypes of renal carcinoma were negative for HLA-G expression. Developing or adult normal renal tissue were devoid of HLA-G expression. We also observed that ectopic expression of HLA class II antigens occurs more frequently in clear cell renal carcinoma than in other subtypes of renal tumors. Moreover, in contrast to the common observation of a down-regulation of major histocompatibility complex class Ia antigens reported in various tumors, the concomitant study of the same biopsies for classical HLA class Ia antigen expression revealed a general increase of HLA class Ia expression, regardless of histological subtypes. These results provide evidence for the heterogeneity of major histocompatibility complex expression patterns in renal carcinoma and support the hypothesis that specific mechanisms underlying the malignant transformation into clear cell renal carcinoma up-regulate expression of HLA-G and to a lesser extent HLA class II molecule expression. Considering the immunotolerant role of HLA-G toward the immune response, these mechanisms may thus provide renal cell carcinoma tumor cells with additional means to escape immune surveillance. PMID- 12547709 TI - Neutrophils mediate parenchymal tissue necrosis and accelerate the rejection of complete major histocompatibility complex-disparate cardiac allografts in the absence of interferon-gamma. AB - A major feature of acute rejection of cardiac allografts is an intense mononuclear cell infiltration accompanied by interferon (IFN)-gamma production. In the current study we tested the role of IFN-gamma in acute rejection of allografts by comparing the histopathology of rejection in wild-type versus IFN gamma-/- recipients of major histocompatibility complex-mismatched cardiac grafts. Wild-type recipients rejected the allografts at days 8 to 9 after transplant but rejection was accelerated 2 to 3 days in IFN-gamma-deficient recipients. During rejection in wild-type recipients, the allografts were heavily infiltrated with CD8+ T cells and other mononuclear cells. In contrast, allografts in IFN-gamma-deficient recipients had few T cells but an intense neutrophil infiltration accompanied by extensive graft parenchymal necrosis. No difference in expression levels of neutrophil chemoattractants including Groalpha/KC, MIP-2, GCP-2, and MIP-1alpha, was observed in allografts retrieved from wild-type and IFN-gamma-/- recipients. Depletion of neutrophils from IFN gamma-deficient recipients delayed rejection until days 8 to 10 after transplant and restored the histopathology of acute allograft rejection to that observed in allografts rejected by wild-type recipients. These results indicate the potent regulatory properties of IFN-gamma during acute rejection directed at neutrophil infiltration into allografts and mediating graft tissue necrosis. PMID- 12547710 TI - Distinct transcriptional profiles of adrenocortical tumors uncovered by DNA microarray analysis. AB - Comprehensive expression profiling of tumors using DNA microarrays has been used recently for molecular classification and biomarker discovery, as well as a tool to identify and investigate genes involved in tumorigenesis. Application of this approach to a cohort of benign and malignant adrenocortical tissues would be potentially informative in all of these aspects. In this study, we generated transcriptional profiles of 11 adrenocortical carcinomas (ACCs), 4 adrenocortical adenomas (ACAs), 3 normal adrenal cortices (NCs), and 1 macronodular hyperplasia (MNH) using Affymetrix HG_U95Av2 oligonucleotide arrays representing approximately 10,500 unique genes. The expression data set was used for unsupervised hierarchical cluster analysis as well as principal component analysis to visually represent the expression data. An analysis of variance on the three classes (NC, ACA plus MNH, and ACC) revealed 91 genes that displayed at least threefold differential expression between the ACC cohort and both the NC and ACA cohorts at a significance level of P < 0.01. Included in these 91 genes were those known to be up-regulated in adrenocortical tumors, such as insulin like growth factor (IGF2), as well as novel differentially expressed genes such as osteopontin (SPP) and serine threonine kinase 15 (STK15). Increased expression of IGF2 was identified in 10 of 11 ACCs (90.9%) and was verified by quantitative reverse transcriptase-polymerase chain reaction. Select proliferation-related genes (TOP2A and Ki-67) were validated at the protein level using immunohistochemistry and adrenocortical tissue microarrays. Our results demonstrated significant and consistent gene expression changes in ACCs compared to benign adrenocortical lesions. Moreover, we identified several genes that represent potential diagnostic markers and may play a role in the pathogenesis of ACC. PMID- 12547712 TI - Down-regulation of sonic hedgehog expression in pulmonary hypoplasia is associated with congenital diaphragmatic hernia. AB - The pathogenesis of pulmonary hypoplasia associated with congenital diaphragmatic hernia (CDH) is unknown. The sonic hedgehog (Shh) cascade is crucial for the patterning of the early respiratory system in mice. To establish whether Shh plays a role in the pathogenesis of lung hypoplasia in CDH, we investigated the gestation-specific expression of Shh in normal rat and human lungs using in situ hybridization and immunohistochemistry. The expression pattern was compared with that of age-matched samples of hypoplastic lungs associated with CDH in humans and in the 2,4-dichlorophenyl-p-nitrophenylether (nitrofen) rat model. Our results showed that in normal controls the expression of Shh increased with advancing gestation, peaked in the late pseudoglandular stage, and declined thereafter. The expression of Shh is initially down-regulated in pulmonary hypoplasia associated with CDH and peaks instead during the late canalicular stage. These data indicate that maximal expression of Shh occurs when respiratory bronchioles develop and thinning of the interstitium takes place, suggesting that Shh may play a role in these processes. Furthermore, we observed that Shh inhibited fetal lung fibroblast proliferation in vitro. Therefore, it is tempting to speculate that alterations in Shh expression may affect these developmental processes, thereby contributing to the pulmonary abnormality in CDH. PMID- 12547711 TI - Global expression profiling of fibroblast responses to transforming growth factor beta1 reveals the induction of inhibitor of differentiation-1 and provides evidence of smooth muscle cell phenotypic switching. AB - Transforming growth factor-beta1 (TGF-beta1) plays a central role in promoting extracellular matrix protein deposition by promoting the transformation of fibroblasts to myofibroblasts. To gain new insights into the transcriptional programs involved, we profiled human fetal lung fibroblast global gene expression in response to TGF-beta1 up to 24 hours using oligonucleotide microarrays. In this report, we present data for 146 genes that were up-regulated at least twofold at two time points. These genes group into several major functional categories, including genes involved in cytoskeletal reorganization (n = 30), matrix formation (n = 25), metabolism and protein biosynthesis (n = 27), cell signaling (n = 21), proliferation and survival (n = 13), gene transcription (n = 9), and of uncertain function (n = 21). For 80 of these genes, this is the first report that they are TGF-beta1-responsive. The early induction of two members of the inhibitor of differentiation (ID) family of transcriptional regulators, ID1 and ID3, was followed by the up-regulation of a number of genes that are usually expressed by highly differentiated smooth muscle cells, including smooth muscle myosin heavy chain, basic calponin, and smoothelin. These findings were confirmed at the protein level for primary adult lung fibroblasts. ID1 further behaved like a typical immediate-early gene and, unlike ID3, was expressed and induced at the protein level. Immunohistochemical analysis showed that ID1 was highly expressed by (myo)fibroblasts within fibrotic foci in experimentally induced pulmonary fibrosis. ID1 acts as a dominant-negative antagonist of basic helix-loop-helix transcription factors that drive cell lineage commitment and differentiation. These findings have important implications for our understanding of fibroblast transcriptional programming in response to TGF-beta1 during development, oncogenesis, tissue repair, and fibrosis. PMID- 12547713 TI - Endothelin-1 suppresses plasma membrane Ca++-ATPase, concomitant with contraction of hepatic sinusoidal endothelial fenestrae. AB - Intracytoplasmic free calcium ions (Ca++) are maintained at a very low concentration in mammalian tissue by extruding Ca++ from the cytoplasm against a steep extracellular Ca++ concentration gradient, mainly through the activity of plasma membrane Ca++ pump-ATPase. The present study aimed to elucidate how endothelin-1 (ET-1) affects the morphology of sinusoidal endothelial fenestrae and ultrastructural distribution of plasma membrane ATPases and intracytoplasmic free Ca++ in isolated rat hepatic sinusoidal endothelial cells. Sinusoidal endothelial fenestrae were observed by scanning electron microscope. Ando's electron cytochemical method was used for ultrastructural localization of Ca++ Mg++-ATPase activity, electron immunogold postembedding method for Ca++ pump ATPase immunoactivity, and antimonate method for intracytoplasmic free Ca++. Addition of ET-1 to sinusoidal endothelial cells significantly decreased Ca++ Mg++-ATPase activity and Ca++ pump-ATPase expression and increased intracytoplasmic free Ca++ concentration, concomitant with a decrease in diameter of sinusoidal endothelial fenestrae. Co-treatment with Bosentan abolished the actions of ET-1. These results suggest that ET-1 suppresses Ca++-Mg++-ATPase activity and Ca++ pump-ATPase expression on the plasma membrane of sinusoidal endothelial fenestrae, thereby attenuating the extrusion of intracytoplasmic free Ca++ into the extracellular space, leading to an increased concentration of intracytoplasmic free calcium ions and contraction of sinusoidal endothelial fenestrae. PMID- 12547714 TI - The role of ultraviolet irradiation and heparin-binding epidermal growth factor like growth factor in the pathogenesis of pterygium. AB - Ultraviolet (UV) light is one of the major factors implicated in the pathogenesis of pterygium. The mechanism by which UV light induces this disease remains elusive. The aim of this study was to evaluate the effects of UVB irradiation on the expression of growth factors in cultured pterygium epithelial cells and to demonstrate their distribution within pterygium. We cultured pterygial epithelial cells from pterygium explants and these cells were exposed to 20 mJ/cm(2) of UVB. Total RNA was extracted at 0, 6, and 12 hours after irradiation. (32)P-labeled cDNA was synthesized and analyzed using microarray technology to determine the differential expression of 268 growth factor and cytokine related genes. Semiquantitative reverse transcriptase-polymerase chain reaction was used to corroborate this data. Conditioned media derived from cells exposed to UVB irradiation was analyzed for protein expression by enzyme-linked immunosorbent assay. Immunohistochemistry was used to evaluate the distribution of heparin binding epidermal growth factor-like growth factor (HB-EGF) in pterygium tissue. Analysis of the hybridization signals revealed that the genes encoding HB-EGF, fibroblast growth factor 3, and cytotoxic trail ligand receptor were consistently elevated at 6 and 12 hours after UVB treatment. HB-EGF mRNA was elevated 6.8-fold at 6 hours after irradiation and was augmented in culture supernatants after the same treatment. Furthermore, HB-EGF reactivity was identified in the epithelium and vasculature of pterygium by immunohistochemistry. HB-EGF was present in normal limbal epithelium, although it was not induced in cultured limbal epithelial cells by UV irradiation. HB-EGF is a potent mitogen, localized in pterygium tissue, and significantly induced by UVB in pterygium-derived epithelial cells. We postulate that this growth factor is a major driving force in the development of pterygia and a means by which UV irradiation causes the pathogenesis of pterygium. PMID- 12547715 TI - Identification of vascular lineage-specific genes by transcriptional profiling of isolated blood vascular and lymphatic endothelial cells. AB - In mammals, the lymphatic vascular system develops by budding of lymphatic progenitor endothelial cells from embryonic veins to form a distinct network of draining vessels with important functions in the immune response and in cancer metastasis. However, the lineage-specific molecular characteristics of blood vascular versus lymphatic endothelium have remained poorly defined. We isolated lymphatic endothelial cells (LECs) and blood vascular endothelial cells (BVECs) by immunomagnetic isolation directly from human skin. Cultured LECs but not BVECs expressed the lymphatic markers Prox1 and LYVE-1 and formed LYVE-1-positive vascular tubes after implantation in vivo. Transcriptional profiling studies revealed increased expression of several extracellular matrix and adhesion molecules in BVECs, including versican, collagens, laminin, and N-cadherin, and of the growth factor receptors endoglin and vascular endothelial growth factor receptor-1/Flt-1. Differential immunostains of human skin confirmed the blood vessel-specific expression of these genes. During embryonic development, endoglin expression was gradually down-regulated on lymphatic endothelium whereas vascular endothelial growth factor receptor-1 was absent from lymphatics. We also identified several genes with specific expression in LECs. These results demonstrate that some lineage-specific genes are only expressed during distinct developmental stages and they identify new molecular markers for blood vascular and lymphatic endothelium with important implications for future studies of vascular development and function. PMID- 12547716 TI - Expression and DNA-binding activity of signal transducer and activator of transcription 3 in alcoholic cirrhosis compared to normal liver and primary biliary cirrhosis in humans. AB - In rats, activation of the cytokine-inducible transcription factor signal transducer and activator of transcription 3 (Stat3) is impaired in the liver after ethanol administration. The aim was to examine Stat3 expression, localization, and activity in alcoholic liver disease (ALD) in humans. Explanted livers of ALD patients were compared to normal and primary biliary cirrhosis livers. Protein expression, DNA-binding, and subcellular localization of Stat3 was examined by Western blotting, electrophoretic mobility shift assays, and immunohistochemistry; and interleukin-6, Stat3, and suppressor of cytokine signaling (SOCS)-3 mRNA expression by quantitative polymerase chain reaction. Stat3 proteins increased markedly in ALD, mainly in hepatocyte and proliferating biliary epithelial cell nuclei. In contrast to normal and primary biliary cirrhosis livers where Stat3 DNA-binding occurred normally, no Stat3 DNA-binding complexes were observed in ALD, although the tyrosine and serine phosphorylation of Stat3 was not altered. Elevated interleukin-6 mRNA was found in ALD whereas Stat3 and suppressor of cytokine signaling-3 mRNA levels were decreased. Although end-stage ALD is characterized by up-regulation of Stat3 proteins, this transcription factor appears to be functionally inactive. Furthermore, decreased transcription of the Stat3 gene in ALD might also affect cytoplasmic reserves of inactivated Stat3 in the long term. Impaired activation and restoration of Stat3 might thus contribute to the development of cell damage leading to liver cirrhosis in ALD. PMID- 12547717 TI - Transforming growth factor-beta mediates intestinal healing and susceptibility to injury in vitro and in vivo through epithelial cells. AB - In vitro studies suggest that transforming growth factor (TGF)-beta has potent effects on gastrointestinal mucosal integrity, wound repair, and neoplasia. However, the multiplicity of actions of this peptide on many different cell types confounds efforts to define the role of TGF-beta within the intestinal epithelium in vivo. To delineate these effects selective blockade of intestinal epithelial TGF-beta activity was undertaken through targeted expression of a dominant negative (DN) TGF-beta RII to intestinal epithelial cells in vitro and in vivo. Stable intestinal epithelial cell (IEC)-6 lines overexpressing TGF-beta RII-DN (nucleotides -7 to 573) were established. Transgenic mice overexpressing TGF-beta RII-DN under the regulation of a modified liver fatty acid-binding promoter (LFABP-PTS4) were constructed. In vitro healing was assessed by wounding of confluent monolayers. Colitis was induced by the addition of dextran sodium sulfate (2.5 to 7.5% w/v) to their drinking water. Overexpression of TGF-beta RII DN in intestinal epithelial cell-6 cells resulted in a marked reduction in cell migration and TGF-beta-stimulated wound healing in vitro. TGF-beta RII-DN transgenic mice did not exhibit baseline intestinal inflammation or changes in survival, body weight, epithelial cell proliferation, aberrant crypt foci, or tumor formation. TGF-beta RII-DN mice were markedly more susceptible to dextran sodium sulfate-induced colitis and exhibited impaired recovery after colonic injury. TGF-beta is required for intestinal mucosal healing and TGF-beta modulation of the intestinal epithelium plays a central role in determining susceptibility to injury. PMID- 12547718 TI - Tumor suppressor role of KiSS-1 in bladder cancer: loss of KiSS-1 expression is associated with bladder cancer progression and clinical outcome. AB - The expression profiles of nine bladder cancer cell lines were compared against a pool containing equal total RNA quantities of each of them. Lower expression of KiSS-1 was revealed in cells derived from the most advanced bladder tumors. When comparing 15 primary bladder tumors versus a pool of four bladder cancer cell lines, lower transcript levels of KiSS-1 were observed in the invasive bladder carcinomas as compared to superficial tumors. KiSS-1 expression ratios provided prognostic information. The expression pattern of KiSS-1 transcripts was analyzed using in situ hybridization in nine bladder cancer cells, paired normal urothelium and bladder tumor samples (n = 25), and tissue microarrays of bladder tumors (n = 173). We observed complete loss of KiSS-1 in all invasive tumors under study as compared to their respective normal urothelium. The expression of KiSS-1 was found to be significantly associated with histopathological stage. Patients with lower KiSS-1 expression showed a direct correlation with overall survival in a subset of bladder tumors whose follow-up was available (n = 69). We did not observe any significant differential KiSS-1 expression along cell cycle by sorting analysis. A potential tumor suppressor role in bladder cancer was revealed for KiSS-1. Moreover, it showed predictive value by identifying patients with poor outcome. PMID- 12547719 TI - A urokinase-derived peptide (A6) increases survival of mice bearing orthotopically grown prostate cancer and reduces lymph node metastasis. AB - The high rate of prostate cancer mortality invariably reflects the inability to control the spread of the disease. The urokinase-type plasminogen activator and its receptor (u-PAR) contribute to prostate cancer metastases by promoting extracellular matrix degradation and growth factor activation. The current study was undertaken to determine the efficacy of a urokinase-derived peptide (A6) in reducing the lymph node metastases of prostate cancer using a model in which prostatic tumors established in nude mice from orthotopically implanted PC-3 LN4 prostate cancer cells disseminate to the lymph nodes. As a first step in evaluating the in vivo effectiveness of A6, we determined its effect on in vitro invasiveness. In vitro, A6 reduced the invasiveness of PC-3 LN4 cells through a Matrigel-coated filter without affecting growth rate. A first in vivo survival experiment showed that all A6-treated mice were alive after 57 days, and half of them tumor-free, whereas all control mice receiving vehicle had died. In a second experiment with a larger tumor inoculum and a longer delay until treatment, whereas 71% of control mice and 83% of mice treated with a scrambled peptide developed lymph node metastases, only 22 to 25% of A6-treated mice had positive lymph nodes. Further, lymph node volume, reflective of tumor burden at the secondary site, was diminished 70% in A6-treated mice. In conclusion, we provide definitive evidence that a peptide spanning the connecting region of urokinase suppresses metastases and, as a single modality, prolongs the life span of prostate tumor-bearing mice. PMID- 12547720 TI - Compromised production of extracellular matrix in mice lacking secreted protein, acidic and rich in cysteine (SPARC) leads to a reduced foreign body reaction to implanted biomaterials. AB - SPARC (secreted protein, acidic and rich in cysteine), a matricellular glycoprotein, modulates the interaction of cells with the extracellular matrix (ECM). Recently, accelerated cutaneous wound closure and altered deposition of collagen were reported in SPARC-null mice. Herein we asked whether SPARC might influence the foreign body reaction to biomaterial implants. Polydimethylsiloxane (silicone rubber) disks and cellulose Millipore filters were implanted into wild type and SPARC-null mice. In wild-type animals, significant levels of SPARC were observed in the cells and the ECM comprising the capsules around the implants. After 4 weeks, SPARC-null mice exhibited a significant decrease in the thickness of the foreign body capsule, as compared to that observed in wild-type mice. A significant reduction in capsular vascular density was also associated with the silicone implants in the SPARC-null animals. Electron microscopy revealed that collagen fibers in the capsules produced by SPARC-null mice were smaller and more uniform in size than those in wild-type animals. Furthermore, staining with picrosirius-red showed that the collagen fibers were less mature in SPARC-null than in wild-type mice. The altered ECM resulting in decreased capsular thickness, indicative of an altered foreign body reaction in SPARC-null mice, implicates SPARC as an important modulator of the encapsulation of implanted biomaterials. PMID- 12547721 TI - Differences in apoptotic susceptibility of cytotrophoblasts and syncytiotrophoblasts in normal pregnancy to those complicated with preeclampsia and intrauterine growth restriction. AB - Placental apoptosis is increased in vivo in preeclampsia (PE) and intrauterine growth restriction (IUGR). The cause and pathological implications of this phenomenon are unknown. This study considers the apoptotic susceptibility of villous trophoblasts from normal, PE, and IUGR pregnancies. Cultured cytotrophoblasts (CTs) and an in vitro model of syncytialization were used. CTs were isolated from term placentas of 12 normal, 12 PE, and 12 IUGR pregnancies. Apoptosis was determined by terminal dUTP nick-end labeling (TUNEL), Annexin V binding, and ADP:ATP ratios. Cells were stimulated with tumor necrosis factor alpha/interferon-gamma or reduced oxygen (<5 KPa). For CTs, ADP:ATP <1 correlates with Annexin V binding. For normal pregnancy, tumor necrosis factor-alpha and depleted oxygen significantly increased TUNEL, Annexin V binding and ADP:ATP in CTs and syncytiotrophoblasts (STs). Spontaneous apoptosis was similar between groups for both cell types. After stimulation, TUNEL and Annexin V binding of CTs were significantly raised in PE and IUGR as compared with normal pregnancy. After oxygen reduction, ADP:ATP in CTs and STs were significantly elevated in IUGR. TUNEL was also increased in STs in PE after oxygen depletion and was significantly raised in STs from IUGR pregnancies after stimulation with both agonists. This is the first description of enhanced apoptosis in isolated villous trophoblasts in PE and IUGR. These intrinsic differences may represent an important factor in the pathophysiology of these conditions. PMID- 12547722 TI - Ghrelin in fetal thyroid and follicular tumors and cell lines: expression and effects on tumor growth. AB - Ghrelin, a growth hormone-releasing hormone produced by gastroenteropancreatic endocrine cells, hypothalamus, and pituitary, was recently identified in medullary thyroid carcinomas and derived cell lines. However, no data exist on its expression in either normal or neoplastic thyroid follicular cells. We analyzed ghrelin expression by immunohistochemistry, in situ hybridization, and reverse transcriptase-polymerase chain reaction in 15 fetal, 4 infant, and 10 adult thyroids, and in 54 tumors of follicular origin. We also analyzed the effects of ghrelin on cell proliferation in N-PAP and ARO thyroid carcinoma cell lines. Ghrelin-binding sites were investigated using reverse transcriptase polymerase chain reaction to detect its growth hormone secretagogue receptor (GHS R) mRNA and an in situ-binding localization procedure. Strong ghrelin immunoreactivity was found in fetal but not in infant or adult thyroids. Ghrelin protein and mRNA were present, in variable amounts, in benign and malignant tumors. Normal thyroids, thyroid tumors, and cell lines showed ghrelin binding sites by binding localization, in the absence of the specific GHS receptor mRNA (with the exception of one normal thyroid). Moreover, ghrelin induced dose dependent inhibition of growth in cell lines. In conclusion, ghrelin is expressed in fetal but not in adult thyroid, and is re-expressed in tumors; the presence of ghrelin receptors other than GHS-R in normal and neoplastic adult thyroid is suggested; ghrelin inhibits cell proliferation of thyroid carcinoma cell lines in vitro. PMID- 12547723 TI - Accelerated in vivo growth of prostate tumors that up-regulate interleukin-6 is associated with reduced retinoblastoma protein expression and activation of the mitogen-activated protein kinase pathway. AB - Interleukin-6 (IL-6) is a multifunctional cytokine that activates the signaling pathways of Janus kinases-signal transducers and activators of transcription (STAT) and/or mitogen-activated protein kinases (MAPK) in various tumors. Thus, it modulates cell growth and apoptosis. IL-6 levels are elevated in tissues and sera from prostate cancer patients and IL-6 receptor expression has been detected in prostate cancer cell lines and clinical specimens. Continuous exposure of prostate cancer cells to IL-6 might alter their responsiveness to this cytokine. To gain more insight into the function of IL-6 in prostate carcinoma, we have inoculated LNCaP-IL-6+ cells, generated after prolonged treatment with IL-6, into nude mice (total n = 16, two independent experiments). Controls included animals bearing LNCaP-IL-6- cells, passaged at the same time as LNCaP-IL-6+ cells without supplementation of IL-6. LNCaP-IL-6+ tumor volumes were larger than those of their counterparts at all time points. There were no signs of cachexia in any of the experimental animals and all mice were free of metastases. To better understand the mechanisms responsible for accelerated growth of LNCaP-IL-6+ tumors, we have investigated the expression of cell-cycle regulatory molecules by Western blot analysis. The levels of cyclin-dependent kinase 2 were elevated in LNCaP-IL-6+ cells. There was a strong down-regulation of cyclins D1 and E in the LNCaP-IL-6+ subline. The cell-cycle inhibitor p27 was expressed at a low level in LNCaP-IL-6+ cells and could not be up-regulated by addition of IL-6. Most notably, LNCaP-IL-6+ cells exhibited a reduced expression of the hypophosphorylated form of the retinoblastoma protein (pRb). Accelerated tumor growth in our model system was also associated with alterations in IL-6-signaling pathways. The ability of IL-6 to induce tyrosine phosphorylation of STAT3 was abolished in the LNCaP-IL-6+ subline. In contrast, the levels of the MAPK extracellular signal-regulated kinases 1/2 increased in cells generated after long-term IL-6 treatment. The inhibitor of MAPK kinase PD 98059 retarded the proliferation of LNCaP-IL-6+ but not that of control cells. In summary, we show in the present study that chronic exposure of prostate cancer cells to IL-6 facilitates tumor growth in vivo by abolishment of the growth control by pRb and activation of the MAPK signaling pathway. These findings could be relevant to understand the role of IL-6 in prostate cancer progression. PMID- 12547724 TI - DNA fingerprinting abnormalities can distinguish ulcerative colitis patients with dysplasia and cancer from those who are dysplasia/cancer-free. AB - Patients with extensive ulcerative colitis (UC) of longer than 8 years duration are at high risk for the development of colorectal cancer. The cancers in these patients appear to develop in a stepwise manner with progressive histological changes from negative for dysplasia --> indefinite for dysplasia --> dysplasia - > cancer. The aim of this study was to determine the timing and extent of genomic instability in the progression of UC dysplasia and cancer. Using two polymerase chain reaction (PCR)-based DNA fingerprinting methods, arbitrarily primed PCR and intersimple sequence repeat PCR, we assessed DNA sequence variation in biopsies across the spectrum of cancerous, dysplastic, and nondysplastic mucosa. UC patients with dysplasia/cancer had substantial genomic instability in both their dysplastic and nondysplastic colonic mucosa, whereas instability was not present in the majority of UC patients without dysplasia/cancer. The degree of instability in nondysplastic tissue was similar to that of dysplastic/cancerous mucosa from the same patient, suggesting that this instability was widespread and reached the maximum level early in neoplastic progression. These results suggest that UC patients who develop dysplasia or cancer have an underlying process of genomic instability in their colonic mucosa whereas UC patients who are dysplasia free do not. PMID- 12547725 TI - Retinal dehydrogenase-2 is inhibited by compounds that induce congenital diaphragmatic hernias in rodents. AB - Currently, the etiology of the serious developmental anomaly congenital diaphragmatic hernia (CDH) is unknown. We have used an animal model of CDH to address this issue. We characterized four separate teratogens that produced diaphragmatic defects in embryonic rats that are similar to those in infants with CDH. We then tested the hypothesis that all these agents share the common mechanism of perturbing the retinoid-signaling pathway. Specifically, inhibition of retinal dehydrogenase-2 (RALDH2), a key enzyme necessary for the production of retinoic acid and that is expressed in the developing diaphragm, was assayed by measuring retinoic acid production in cytosolic extracts from an oligodendrocyte cell line. The following compounds all induce posterolateral defects in the rat diaphragm; nitrofen, 4-biphenyl carboxylic acid, bisdiamine, and SB-210661. Importantly, we demonstrate that they all share the common mechanism of inhibiting RALDH2. These data provide an important component of mounting evidence suggesting that the retinoid system warrants consideration in future studies of the etiology of CDH. PMID- 12547726 TI - Splenic marginal zone lymphoma with villous lymphocytes shows on-going immunoglobulin gene mutations. AB - Splenic marginal zone lymphoma (also splenic lymphoma with villous lymphocytes) is a B-cell non-Hodgkin's lymphoma with a characteristic morphology and phenotype. We studied the pattern of somatic hypermutation of the rearranged immunoglobulin heavy chain genes on 23 cases and have correlated these data with survival as well as immunophenotypic and genetic characteristics of the cases. Two-thirds of the cases show immunoglobulin gene mutations, half of which show evidence of antigen selection, whereas one-third of the cases show no significant mutations. On-going mutation, a feature characteristic of follicular lymphoma, was demonstrated in all six cases randomly selected for this analysis, including one case with a low number of mutations (<2%). No statistical significant correlation was found between immunoglobulin mutation status and clinical, immunophenotypic, or genetic characteristics. Our results demonstrate that on going somatic hypermutation is a prominent feature of splenic marginal zone lymphoma with circulating villous lymphocytes. On-going somatic hypermutation has previously been demonstrated in extra-nodal and nodal marginal zone lymphoma. Our results indicate that marginal zone lymphomas at different anatomical localizations may derive from a similar B-cell subset. PMID- 12547727 TI - CD4+ CD25+ regulatory T lymphocytes inhibit microbially induced colon cancer in Rag2-deficient mice. AB - Inflammatory bowel diseases, including ulcerative colitis and Crohn's disease, increase the risk of colorectal cancer in humans. It has been recently shown in humans and animal models that intestinal microbiota and host immunity are integral in the progression of large bowel diseases. Lymphocytes are widely believed to prevent bacterially induced inflammation in the bowel, and lymphocytes are also critical in protecting against primary tumors of intestinal epithelia in mice. Taken together, this raises the possibility that lymphocytes may inhibit colon carcinogenesis by reducing bacterially driven inflammation. To examine the role of bacteria, lymphocytes, and inflammatory bowel disease in the development of colon cancer, 129/SvEv Rag-2-deficient and congenic wild-type mice were orally inoculated with a widespread enteric mouse bacterial pathogen, Helicobacter hepaticus, or sham-dosed with media only. H. hepaticus-infected Rag2 /-, but not sham-dosed Rag2-/- mice, rapidly developed colitis and large bowel carcinoma. This demonstrated a link between microbially driven inflammation and cancer in the lower bowel and suggested that innate immune dysregulation may have an important role in inflammatory bowel disease and progression to cancer. H. hepaticus-infected wild-type mice did not develop inflammation or carcinoma showing that lymphocytes were required to prevent bacterially induced cancer at this site. Adoptive transfer with CD4+ CD45RBlo CD25+ regulatory T cells into Rag deficient hosts significantly inhibited H. hepaticus-induced inflammation and development of cancer. These results suggested that the ability of CD4+ T cells to protect against intestinal cancer was correlated with their ability to reduce bacterially induced inflammatory bowel disease. Further, regulatory T cells may act directly on the innate immune system to reduce or prevent disease. These roles for T cells in protection against colon carcinoma may have implications for new modes of prevention and treatment of cancer in humans. PMID- 12547728 TI - Distinct cytokine production by lung and blood neutrophils from children with cystic fibrosis. AB - Inflammation plays a critical role in lung disease progression in cystic fibrosis (CF). This inflammatory process is dominated by a neutrophil influx in the airways. To determine whether the accumulation of neutrophils in the airways of CF patients is associated with an altered function, we analyzed the capacity of neutrophils isolated from the lung compartment and the blood to release the major neutrophil pro- and anti-inflammatory cytokines IL-8 and IL-1-receptor antagonist (ra) spontaneously and in the presence of LPS. Comparison of cytokine production by blood neutrophils from CF patients and from control subjects showed significantly increased IL-8 and decreased IL-1ra release by CF neutrophils. Comparison of cytokine production by airway and blood neutrophils from CF patients also documented distinct profiles: the spontaneous release of IL-8 and IL-1ra by airway neutrophils was significantly higher than that from blood neutrophils. Culture in the presence of LPS failed to further enhance cytokine production. Analysis of the effect of dexamethasone confirmed the difference in the responsiveness of lung and blood neutrophils in CF. Used at a concentration effective in reducing IL-8 production by blood neutrophils, dexamethasone (10(-6) M) was unable to repress secretion of IL-8 by airway neutrophils. In addition, comparison of cytokine production by airway neutrophils from children with CF and children with dyskinetic cilia syndrome also documented distinct profiles of secretion. These results are consistent with a dysregulated cytokine production by lung and blood neutrophils in CF. They provide support to the hypothesis that not only the CF genotype but also the local environment may modify the functional properties of the neutrophils. PMID- 12547729 TI - Role of VEGF-B in the lung during development of chronic hypoxic pulmonary hypertension. AB - Angiogenic factors exert protective effects on the lung. To investigate the effect of VEGF-B, a factor coexpressed in the lung with VEGF-A, we assessed chronic hypoxic pulmonary hypertension in VEGF-B knockout mice (VEGF-B-/-) and in rats with lung overexpression of VEGF-B induced by adenovirus transfer. No significant difference in pulmonary hemodynamics, right ventricular hypertrophy, distal vessel muscularization, or vascular density was found between VEGF-B-/- and control mice after 3 wk of hypoxia. When overexpressed, VEGF-B(167) or VEGF B(186) had protective effects similar to those of human VEGF-A(165). Lung endothelial nitric oxide synthase (eNOS) expression was increased by 5 days of hypoxia or VEGF-A adenovirus vector (Ad.VEGF-A) overexpression, whereas VEGF B(167) or VEGF-B(186) had no effect. With hypoxia or normoxia, the wet-to-dry lung weight ratio was increased 5 days after Ad.VEGF-A administration compared with control (Ad.nul), Ad.VEGF-B(167), or Ad.VEGF-B(186). Endogenous VEGF-B does not counteract the development of hypoxic pulmonary hypertension. However, when overexpressed in the lung, VEGF-B can be as potent as VEGF-A in attenuating pulmonary hypertension, although it has no effect on eNOS expression or vascular permeability. PMID- 12547730 TI - cAMP activates BKCa channels in pulmonary arterial smooth muscle via cGMP dependent protein kinase. AB - The signal transduction mechanisms defining the role of cyclic nucleotides in the regulation of pulmonary vascular tone is currently an area of great interest. Normally, signaling mechanisms that elevate cAMP and guanosine-3',5'-cyclic monophosphate (cGMP) maintain the pulmonary vasculature in a relaxed state. Modulation of the large-conductance, calcium- and voltage-activated potassium (BK(Ca)) channel is important in the regulation of pulmonary arterial pressure, and inhibition (closing) of the BK(Ca) channel has been implicated in the development of pulmonary hypertension. Accordingly, studies were done to determine the effect of cAMP-elevating agents on BK(Ca) channel activity using patch-clamp studies in pulmonary arterial smooth muscle cells (PASMC) of the fawn hooded rat (FHR), a recognized animal model of pulmonary hypertension. Forskolin (10 micro M), a stimulator of adenylate cyclase and an activator of cAMP dependent protein kinase (PKA), and 8-4-chlorophenylthio (CPT)-cAMP (100 micro M), a membrane-permeable derivative of cAMP, opened BK(Ca) channels in single FHR PASMC. Treatment of FHR PASMC with 300 nM KT5823, a selective inhibitor of cGMP dependent protein kinase (PKG) activity inhibited the effect of both forskolin and CPT-cAMP. In contrast, blocking PKA activation with 300 nM KT5720 had no effect on forskolin or CPT-cAMP-stimulated BK(Ca) channel activity. These results indicate that cAMP-dependent vasodilators activate BK(Ca) channels in PASMC of FHR via PKG-dependent and PKA-independent signaling pathways, which suggests cross-activation between cyclic nucleotide-dependent protein kinases in pulmonary arterial smooth muscle and therefore, a unique signaling pathway for cAMP-induced pulmonary vasodilation. PMID- 12547731 TI - Pulmonary lymphatics and edema accumulation after brief lung injury. AB - In a past study of hyperoxia-induced lung injury, the extensive lymphatic filling could have resulted from lymphatic proliferation or simple lymphatic recruitment. This study sought to determine whether brief lung injury could produce similar changes, to show which lymphatic compartments fill with edema, and to compare their three-dimensional structure. Tracheostomized rats were ventilated at high tidal volume (12-16 ml) or low tidal volume (3-5 ml) or allowed to breathe spontaneously for 25 min. Light microscopy showed more perivascular, interlobular septal, and alveolar edema in the animals ventilated at high tidal volume (P < 0.0001). Scanning electron microscopy of lymphatic casts showed extensive filling of the perivascular lymphatics in the group ventilated at high tidal volume (P < 0.01), but lymphatic filling was greater in the nonventilated group than in the group that was ventilated at low tidal volume (P < 0.01). The three-dimensional structures of the cast interlobular and perivascular lymphatics were similar. There was little filling and no difference in pleural lymphatic casts among the three groups. More edema accumulated in the surrounding lymphatics of larger blood vessels than smaller blood vessels. Brief high-tidal-volume lung injury caused pulmonary edema similar to that caused by chronic hyperoxic lung injury, except it was largely restricted to perivascular and septal lymphatics and prelymphatic spaces. PMID- 12547732 TI - What leads to different mediators of alkalosis-induced vasodilation in isolated and in situ pulmonary vessels? AB - We previously found that nitric oxide synthase (NOS) inhibition fully blocked alkalosis-induced relaxation of piglet pulmonary artery and vein rings. In contrast, NOS inhibition alone had no effect on alkalosis-induced pulmonary vasodilation in isolated piglet lungs. This study sought to identify factors contributing to the discordance between isolated and in situ pulmonary vessels. The roles of pressor stimulus (hypoxia vs. the thromboxane mimetic U-46619), perfusate composition (blood vs. physiological salt solution), and flow were assessed. Effects of NOS inhibition on alkalosis-induced dilation were also directly compared in 150-350-microm-diameter cannulated arteries and 150-900 microm-diameter, angiographically visualized, in situ arteries. Finally, effects of NOS inhibition on alkalosis-induced vasodilation were measured in intact piglets. NOS inhibition with N(omega)-nitro-L-arginine fully abolished alkalosis induced vasodilation in all cannulated arteries but failed to alter alkalosis induced vasodilation in intact lungs. The results indicate that investigation of other factors, such as perivascular tissue (e.g., adventitia and parenchyma) and remote signaling pathways, will need to be carried out to reconcile this discordance between isolated and in situ arteries. PMID- 12547733 TI - Cigarette smoke extract induces oxidative stress and apoptosis in human lung fibroblasts. AB - Cigarette smoke is a mixture of chemicals having direct and/or indirect toxic effects on different lung cells. We investigated the effect of cigarette smoke on human lung fibroblasts (HFL-1) oxidation and apoptosis. Cells were exposed to various concentrations (1, 5, and 10%) of cigarette smoke extract (CSE) for 3 h, and oxidative stress and apoptosis were assessed by fluorescence-activated cell sorting and confocal laser fluorescence microscopy. Both oxidative stress and apoptosis exhibited a dose-response relationship with CSE concentrations. Lung fibroblasts also showed marked DNA fragmentation at the Comet assay after exposure to 10% CSE. Coincubation of HLF-1 cells with N-acetylcysteine (1 mM) during CSE exposure significantly reduced oxidative stress, apoptosis, and DNA fragmentation, whereas preincubation (3 h) with the glutathione-depleting agent buthionine sulfoximine (125 microM) produced a significant increase of oxidative stress. Cigarette smoke is a potent source of oxidative stress, DNA damage, and apoptosis for HFL-1 cells, and we speculate that this could contribute to the development of pulmonary emphysema in the lungs of smokers. PMID- 12547735 TI - Computerised neuropsychological testing. PMID- 12547737 TI - Preparticipation screening for the detection of cardiovascular abnormalities that may cause sudden death in competitive athletes. PMID- 12547738 TI - Guidelines of the American College of Obstetricians and Gynecologists for exercise during pregnancy and the postpartum period. PMID- 12547740 TI - Musculoskeletal manifestations of diabetes mellitus. AB - Rheumatic complaints are common in patients with diabetes. Maintaining good glycaemic control by exercise, diet, and medication improves or prevents the development of rheumatic conditions. PMID- 12547741 TI - Incidence of injury in semi-professional rugby league players. AB - OBJECTIVES: To investigate the site, nature, cause, and severity of injuries in semi-professional rugby league players. METHODS: The incidence of injury was prospectively studied in one hundred and fifty six semi-professional rugby league players over two competitive seasons. All injuries sustained during matches and training sessions were recorded. Injury data were collected from a total of 137 matches and 148 training sessions. Information recorded included the date and time of injury, site, nature, cause, and severity of injury. RESULTS: During the two seasons, 1,694 playing injuries and 559 training injuries were sustained. The match injury incidence was 824.7 per 1,000 player-position game hours and training injury incidence was 45.3 per 1,000 training hours. Over 20% of the total training (17.4 per 1,000) and playing (168.0 per 1,000) injuries sustained were to the thigh and calf. Muscular injuries (haematomas and strains) were the most common type of injury sustained during training (22.0 per 1,000, 48.7%) and matches (271.7 per 1,000, 32.9%). Playing injuries were most commonly sustained in tackles (382.2 per 1,000, 46.3%), while overexertion was the most common cause of training injuries (15.5 per 1,000, 34.4%). The majority of playing injuries were sustained in the first half of matches (1,013.6 per 1,000, 61.5% v 635.8 per 1,000, 38.5%), whereas training injuries occurred more frequently in the latter stages of the training session (50.0 per 1,000, 55.3% v 40.5 per 1,000, 44.7%). Significantly more training injuries were sustained in the early half of the season, however, playing injuries occurred more frequently in the latter stages of the season. CONCLUSIONS: These results suggest that changes in training and playing intensity impact significantly upon injury rates in semi-professional rugby league players. Further studies investigating the influence of training and playing intensity on injuries in rugby league are warranted. PMID- 12547739 TI - Risk factors for lower extremity injury: a review of the literature. AB - Prospective studies on risk factors for lower extremity injury are reviewed. Many intrinsic and extrinsic risk factors have been implicated; however, there is little agreement with respect to the findings. Future prospective studies are needed using sufficient sample sizes of males and females, including collection of exposure data, and using established methods for identifying and classifying injury severity to conclusively determine additional risk factors for lower extremity injury. PMID- 12547742 TI - The effect of glucosamine supplementation on people experiencing regular knee pain. AB - OBJECTIVE: The purpose of this study was to examine the effects of oral glucosamine supplementation on the functional ability and degree of pain felt by individuals who had regular knee pain, most likely due to previous articular cartilage damage, and possibly osteoarthritis. METHODS: Subjects were randomly supplemented with either glucosamine (G) (n=24) or placebo (P) (lactose) (n=22) for 12 weeks at a dose of 2,000 mg per day. Over this period, four testing sessions were conducted, with changes in knee pain and function assessed by clinical and functional tests, (joint line palpation, a 3 metre "duck walk" and a repeated, walking stair climb), two questionnaires (the Knee Injury and Osteoarthritis Outcome Score (KOOS) and the Knee Pain Scale (KPS)) and participant subjective evaluations. RESULTS: The clinical and functional test scores improved with time (main effects: p<0.05, p<0.01) but there were no significant differences between the two groups. The questionnaire results also recorded a significant main effect for time (p<0.05), but the glucosamine group was found to have significantly better KOOS quality of life scores at week eight and 12 (p<0.05), and lower KPS scores (p<0.05) at week eight than the placebo group. On self report evaluations of changes across the 12 week supplementation period, 88% (n=21) of the glucosamine group reported some degree of improvement in their knee pain versus only 17% (n=3) in the placebo group. CONCLUSIONS: These results suggest that glucosamine supplementation can provide some degree of pain relief and improved function in persons who experience regular knee pain, which may be caused by prior cartilage injury and/or osteoarthritis. The trends in the results also suggest that, at a dosage of 2,000 mg per day, the majority of improvements are present after eight weeks. PMID- 12547743 TI - The physiological effects of cycling on tandem and single bicycles. AB - OBJECTIVE: The purpose of this field study was to compare the physiological responses from cycling on a tandem road bicycle to those from cycling on a single road bicycle. METHODS: Nine pairs of experienced, recreational tandem cyclists rode a tandem or their single bicycle for 5 min at each velocity of 19.3, 22.5, 25.8, and 29.0 kph on a flat, paved surface. Heart rate (HR), rating of perceived exertion (RPE), and lactic acid (LA) data were collected after each interval. RESULTS: Riding a tandem resulted in lower HR, RPE, and LA mean values across the four velocities compared to the single bicycle. Mean (SD) HR, RPE, and LA for tandem and single bicycles were 126 (20.7) v 142 (20.1) bpm, 10.1 (1.7) v 11.3 (2.6), and 1.46 (1.0) mM/L v 2.36 (1.7) mM/L, respectively. No position differences were observed between the captain and stoker (front and rear positions) when both were on the tandem. Stokers had significantly lower HR, LA, and RPE values when they rode a tandem compared to a single bicycle. No statistical differences were observed between bicycles for the captains. When on the single bicycle, captains exhibited significantly lower HR, RPE, and LA values than stokers. CONCLUSION: Cycling on a tandem resulted in lower physiological stress than when cycling at the same velocity on a single bicycle. Cyclists were able to ride from 4.8-8.0 kph faster on a tandem than on a single bicycle at similar physiological stress. Apparently, stokers can add to power output on a tandem without adding significantly to wind resistance. PMID- 12547744 TI - Changes in muscle strength and pain in response to surgical repair of posterior abdominal wall disruption followed by rehabilitation. AB - BACKGROUND: Posterior abdominal wall deficiency (PAWD) is a tear in the external oblique aponeurosis or the conjoint tendon causing a posterior wall defect at the medial end of the inguinal canal. It is often known as sportsman's hernia and is believed to be caused by repetitive stress. OBJECTIVE: To assess lower limb and abdominal muscle strength of patients with PAWD before intervention compared with matched controls; to evaluate any changes following surgical repair and rehabilitation. METHODS: Sixteen subjects were assessed using a questionnaire, isokinetic testing of the lower limb strength, and pressure biofeedback testing of the abdominals. After surgery and a six week rehabilitation programme, the subjects were re-evaluated. A control group were assessed using the same procedure. RESULTS: Quadriceps and hamstrings strength was not affected by this condition. A deficit hip muscle strength was found on the affected limb before surgery, which was significant for the hip flexors (p = 0.05). Before surgery, 87% of the patients compared with 20% of the controls failed the abdominal obliques test. Both the injured and non-injured sides had improved significantly in strength after surgery and rehabilitation. The strength of the abdominal obliques showed the most significant improvement over the course of the rehabilitation programme. CONCLUSIONS: Lower limb muscle strength may have been reduced as the result of disuse atrophy or pain inhibition. Abdominal oblique strength was deficient in the injured patients and this compromises rotational control of the pelvis. More sensitive investigations (such as electromyography) are needed to assess the link between abdominal oblique function and groin injury. PMID- 12547745 TI - Comparison of three different sit and reach tests for measurement of hamstring flexibility in female university students. AB - BACKGROUND: The sit and reach test is the most common flexibility test used in health related fitness test batteries. OBJECTIVE: To examine and compare three different sit and reach tests as a measure of hamstring flexibility in 102 female students. METHOD: The traditional sit and reach test, the chair sit and reach test, the back saver sit and reach test, and passive straight leg raise were administered in three trials to all 102 students (mean (SD) age 22 (1) years) on the same day. RESULTS: A Pearson correlation coefficient analysis was significant (p<0.01) for the traditional sit and reach test with back saver sit and reach test and flexibility of hamstrings (r = 0.45 and 0.65 for left and right legs, and 0.63 and 0.53 for left and right legs respectively). Also, the back saver sit and reach test for the left (p<0.01) and right (p<0.05) leg was significantly associated with hamstring flexibility (r = 0.37 and 0.25 for the left leg and 0.50 and 0.44 for the right leg respectively). CONCLUSION: The results indicate that the back saver sit and reach test produces reasonably accurate and stable measures of hamstring flexibility. Moreover, it appears that this test is a safe and acceptable alternative to the traditional and chair sit and reach tests as a measure of hamstring flexibility in young women. PMID- 12547746 TI - Analysis of the characteristics of competitive badminton. AB - OBJECTIVE: To describe the characteristics of badminton in order to determine the energy requirements, temporal structure, and movements in the game that indicate performance level. To use the findings to plan training with greater precision. METHODS: Eleven badminton players (mean (SD) age 21.8 (3.26) years) with international experience from four different countries (France, Italy, Spain, and Portugal) were studied. Two of the Spanish players were monitored in several matches, giving a total of 14 samples, all during the 1999 Spanish International Tournament. Blood lactate concentration was measured with a reflective photometer. Maximum and average heart rates were recorded with a heart rate monitor. Temporal structure and actions during the matches were determined from video recordings. All variables were measured during and after the game and later analysed using a descriptive study. RESULTS: The results confirmed the high demands of the sport, with a maximum heart rate of 190.5 beats/min and an average of 173.5 beats/min during matches over 28 minutes long and performance intervals of 6.4 seconds and rest time of 12.9 seconds between exchanges. CONCLUSIONS: The results suggest that badminton is characterised by repetitive efforts of alactic nature and great intensity which are continuously performed throughout the match. An awareness of these characteristics, together with data on the correlations between certain actions such as unforced errors and winning shots and the final result of the match, will aid in more appropriate planning and monitoring of specific training. PMID- 12547747 TI - Physiological factors associated with low bone mineral density in female endurance runners. AB - OBJECTIVE: To explore potential factors that could be associated with low bone mineral density (BMD) in female endurance runners. METHODS: Fifty two female endurance runners (1,500 m to marathon), aged 18-44 years, took part. Body fat percentage, lumbar spine BMD, and femoral neck BMD were measured using the Hologic QDR 4,500w bone densitometer. Data on training, menstrual cycle status, osteoporosis, and health related factors were obtained by questionnaire. Dietary variables were assessed from a prospective seven day dietary record of macronutrients and micronutrients. RESULTS: The mean (SD) lumbar spine and femoral neck BMD were 1.11 (0.11) and 0.89 (0.12) g/cm(2) respectively. A backward elimination regression analysis showed that age, body mass, body fat, distance run, magnesium, and zinc intake were the variables significantly associated with BMD. Lumbar spine BMD (g/cm(2)) = -1.90 + (0.0486 x age (years)) + (0.342 x log mass (kg)) - (0.000861 x age(2) (years)) - (0.00128 x distance (km/week)), with an R(2) = 30.1% (SEE = 0.089 (95% confidence interval (CI) 0.05 to 0.23); p<0.001). Femoral neck BMD (g/cm(2)) = -2.51 - (0.00989 x age (years)) + (0.720 x log mass (kg)) + (0.000951 x magnesium (mg/day)) -(0.0289 x zinc (mg/day)) - (0.00821 x body fat (%)) - (0.00226 x distance (km/week)), with an R(2) = 50.2% (SEE = 0.100 (95% CI 0.06 to 0.22); p<0.001). The negative association between skeletal BMD and distance run suggested that participants who ran longer distances had a lower BMD of the lumbar spine and femoral neck. Further, the results indicated a positive association between body mass and BMD, and a negative association between body fat and BMD. CONCLUSIONS: The results suggest a negative association between endurance running distance and lumbar spine and femoral neck BMD, with a positive association between body mass and femoral neck and lumbar spine BMD. However, longitudinal studies are required to assess directly the effect of endurance running and body mass on BMD, and to see if the addition of alternative exercise that would increase lean body mass would have a positive effect on BMD and therefore help to prevent osteoporosis. PMID- 12547748 TI - The effects of massage on delayed onset muscle soreness. AB - OBJECTIVES: The purpose of this study was to investigate the physiological and psychological effects of massage on delayed onset muscle soreness (DOMS). METHODS: Eighteen volunteers were randomly assigned to either a massage or control group. DOMS was induced with six sets of eight maximal eccentric contractions of the right hamstring, which were followed 2 h later by 20 min of massage or sham massage (control). Peak torque and mood were assessed at 2, 6, 24, and 48 h postexercise. Range of motion (ROM) and intensity and unpleasantness of soreness were assessed at 6, 24, and 48 h postexercise. Neutrophil count was assessed at 6 and 24 h postexercise. RESULTS: A two factor ANOVA (treatment v time) with repeated measures on the second factor showed no significant treatment differences for peak torque, ROM, neutrophils, unpleasantness of soreness, and mood (p > 0.05). The intensity of soreness, however, was significantly lower in the massage group relative to the control group at 48 h postexercise (p < 0.05). CONCLUSIONS: Massage administered 2 h after exercise induced muscle injury did not improve hamstring function but did reduce the intensity of soreness 48 h after muscle insult. PMID- 12547749 TI - Effects of 12 months of exercise training on salivary secretory IgA levels in elderly subjects. AB - BACKGROUND: The immune system declines in efficiency with advancing age, making the elderly less resistant to pathogenic microorganisms. Upper respiratory tract infection (URTI) is a common illness. Recent studies have shown that suppression of secretory immunoglobulin A (SIgA) is associated with increased incidence of URTI. OBJECTIVE: To assess the effect of exercise on salivary SIgA in elderly subjects. METHODS: Forty five elderly subjects (18 men, 27 women; mean (SD) age 64.9 (8.4) years) performed both 60 minute resistance and 60 minute moderate endurance training a week for 12 months. Saliva samples were obtained before training, and at four and 12 months during the training period. Salivary SIgA concentrations were measured by enzyme linked immunosorbent assay, and the SIgA secretion rate was calculated. RESULTS: SIgA concentrations before training, and at four and 12 months during training were 24.7 (14.4), 27.2 (14.2), and 33.8 (18.5) micro g/ml respectively. SIgA secretion rates were 29.5 (26.0), 33.8 (27.2) and 46.5 (35.1) micro g/min respectively. The results indicate that both the concentration and secretion rate of SIgA significantly (p<0.01) increased during 12 months of exercise in these elderly subjects. CONCLUSION: Regular moderate exercise seems to enhance mucosal immune function in elderly subjects. PMID- 12547750 TI - Ex-professional association footballers have an increased prevalence of osteoarthritis of the hip compared with age matched controls despite not having sustained notable hip injuries. AB - OBJECTIVE: To compare the prevalence of self reported, physician diagnosed osteoarthritis (OA) of the hip in ex-professional footballers with that in age matched controls in the general public. METHOD: A questionnaire was sent to the 92 football league and premiership managers to assess the prevalence of OA of various joints. The prevalence of OA of the hip in those managers that were ex professional footballers was assessed. Radiographic controls were obtained and used to assess the prevalence of OA of the hip in the general population. The results of the two groups were compared statistically using chi(2) tabulation. RESULTS: Seventy four (80%) of the managers responded to the questionnaire. Nine of the 68 ex-professional footballers who replied had OA of the hip, and six of these had undergone a total of eight total hip replacements. Of the 18 managers who did not respond, 12 were known to be ex-professionals. In the control group of 136, two had OA of the hip. The two groups differed significantly (p<0.001). The odds ratio for OA of the hip was 10.2 (95% confidence interval 2.1 to 48.8). CONCLUSION: The ex-professional footballers had a significantly higher prevalence of OA of the hip than an age matched group of radiographic controls. PMID- 12547751 TI - Chronic subdural haematoma after snowboard head injury. AB - Two cases of chronic subdural haematoma following a snowboard head injury are reported. Although such cases are rare in sport, the risk in snowboarders is higher than expected. Evaluation of a snowboarder with a history of head injury, albeit mild, who complains of headaches should include computed tomography or magnetic resonance imaging to allow rapid identification of any intracranial pathology. PMID- 12547752 TI - Tear of the acetabular labrum in an elite athlete. AB - The case is reported of a professional footballer with a tear of the acetabular labrum. The clinical findings, treatment, and rehabilitation are described as well as a discussion of the difficulties of diagnosis. PMID- 12547753 TI - Heart rate response to ultraendurance cycling. AB - The heart rate (HR) response to ultraendurance cycling is poorly understood. This case report describes the exercise intensity of ultraendurance cycling by means of HR monitoring in a well trained male amateur cyclist performing the Otztal Radmarathon twice en bloque in a circuit of two identical laps (distance 460 km; cumulative altitude difference 11,000 m). The overall intensity was moderate (HR(mean) = 130 beats/min; HR(mean)/HR(max) = 0.71) corresponding to an average individual workload of 47% of VO(2)MAX. Almost the whole race was performed under aerobic conditions (99.6%); high intensity work was negligible (0.4%). The average speed and the HR response also declined in the course of the two laps, average speed by 17.2% (23.8 to 19.7 km/h), HR(mean) by 10.1% (138 to 124 beats/min), and HR(mean)/HR(max) by 10.7% (0.75 to 0.67). This scale of HR decrease corresponds to comparable data gained in the field of triathlon and represents a specific cardiac feature of ultraendurance exercise in general. PMID- 12547754 TI - Rehabilitation of lumbar multifidus dysfunction in low back pain: strengthening versus a motor re-education model. PMID- 12547755 TI - Increased endothelin-1 levels in athletes. PMID- 12547756 TI - The event side doctor: the role of the orthopaedic surgeon. PMID- 12547757 TI - Relating single-molecule measurements to thermodynamics. AB - Measurements made on large ensembles of molecules are routinely interpreted using thermodynamics, but the normal rules of thermodynamics may not apply to measurements made on single molecules. Using a polymer stretching experiment as an example, it is shown that in the limit of a single, short molecule the outcome of experimental measurements may depend on which variables are held fixed and which are allowed to fluctuate. Thus an experiment in which the end-to-end distance of the polymer molecule is fixed and the tension fluctuates yields a different result than an experiment where the force is fixed and the end-to-end distance fluctuates. It is further shown that this difference is due to asymmetry in the distribution of end-to-end distances for a single molecule, and that the difference vanishes in the appropriate thermodynamic limit; that is, as the polymer molecule becomes long compared to its persistence length. Despite these differences, much of the thermodynamic formalism still applies on the single molecule level if the thermodynamic free energies are replaced with appropriate potentials of mean force. The primary remaining differences are consequences of the fact that unlike the free energies, the potentials of mean force are not in general homogeneous functions of their variables. The basic thermodynamic concepts of an intensive or extensive quantity, and the thermodynamic relationships that follow from them, are therefore less useful for interpreting single-molecule experiments. PMID- 12547758 TI - Limiting frequency of the cochlear amplifier based on electromotility of outer hair cells. AB - Outer hair cells are the critical element for the sensitivity and sharpness of frequency selectivity of the ear. It is believed that fast motility (electromotility) of these cells is essential for this function. Indeed, force produced by outer hair cells follows their membrane potential very closely at least up to 60 kHz. However, it has been pointed out that the cell's receptor potential is attenuated by a low-pass RC circuit inherent to these cells, with the RC roll-off frequencies significantly lower than their operating frequencies. This would render electromotility ineffective in producing force. To address this issue, we assume that multiple degrees of freedom and vibrational modes due to the complex structure of the organ of Corti provide optimal phases for outer hair cells' force to cancel viscous drag. Our derived frequency limit depends on the drag-capacitance product, not directly on the RC time constant. With a reasonable assumption for the viscous drag, the estimated limit is 10-13 kHz, exceeding the RC corner frequency. Our analysis shows that a fast-activating potassium current can substantially extend the frequency limit by counteracting the capacitive current. PMID- 12547759 TI - Prediction of secondary ionization of the phosphate group in phosphotyrosine peptides. AB - A computational approach, based on a continuum molecular electrostatics model, for the calculation of the pK(a) values of secondary ionization of the phosphate group in phenyl phosphate derivatives is described. The method uses the ESP atomic charges of the mono-anionic and di-anionic forms of the ionizable phosphate group, computed with the use of the density functional method, and applies a new concept of the model group, being the reference state for the pK(a) calculations. Both conformational flexibility and tautomeric degrees of freedom are taken into account in the calculations. The method was parameterized using experimentally available pK(a) values of four derivatives of phenyl phosphates, and phosphotyrosine. Subsequently this parameterization was used to predict pK(a) of the phosphate group in a short peptide Gly-Gly-Tyr(P)-Ala, and in a longer peptide consisting of 12 residues, the latter in water, and in a complex with a protein-phospholipase. The agreement between the computed and the experimental pK(a) values is better than +/-0.3 pH units for the optimized solute dielectric constant of 11-13. This approach is promising and its extension to other phospho amino acids is in progress. PMID- 12547760 TI - A force balance model of early spindle pole separation in Drosophila embryos. AB - The formation and function of the mitotic spindle depends upon force generation by multiple molecular motors and by the dynamics of microtubules, but how these force-generating mechanisms relate to one another is unclear. To address this issue we have modeled the separation of spindle poles as a function of time during the early stages of spindle morphogenesis in Drosophila embryos. We propose that the outward forces that drive the separation of the spindle poles depend upon forces exerted by cortical dynein and by microtubule polymerization, and that these forces are antagonized by a C-terminal kinesin, Ncd, which generates an inward force on the poles. We computed the sum of the forces generated by dynein, microtubule polymerization, and Ncd, as a function of the extent of spindle pole separation and solved an equation relating the rate of pole separation to the net force. As a result, we obtained graphs of the time course of spindle pole separation during interphase and prophase that display a reasonable fit to the experimental data for wild-type and motor-inhibited embryos. Among the novel contributions of the model are an explanation of pole separation after simultaneous loss of Ncd and dynein function, and the prediction of a large value for the effective centrosomal drag that is needed to fit the experimental data. The results demonstrate the utility of force balance models for explaining certain mitotic movements because they explain semiquantitatively how the force generators drive a rapid initial burst of pole separation when the net force is great, how pole separation slows down as the force decreases, and how a stable separation of the spindle poles characteristic of the prophase steady state is achieved when the force reaches zero. PMID- 12547761 TI - Competitive model on denaturant-mediated protein unfolding. AB - A denaturant-mediated protein unfolding model, which is different from already existing ones based on the assumption that denaturant competes for water molecules to interact and thus reduces water-protein interactions, which leads to unfolding phenomenon, has been developed with a detailed mathematical justification. Theoretical results suggested that the parameter (m(u)) obtained from the usual linear extrapolation model must be a linear function of the number of bound water molecules (n) on protein with a zero intercept. However, application of this theory to a set of proteins for which m(u) values for urea denaturation are already known showed that m(u) was a linear function of n but with a nonzero intercept. Finally this nonzero intercept was attributed to binding of denaturant to protein at n = 0. Detailed investigation of this factor showed that average equilibrium constant for binding of urea with aromatic side chains (generally nonpolar side chains) was k(b) approximately 0.65 +/- 0.45 mol( 1), which agreed well with earlier experimental estimations, and also suggested that an integrated approach was necessary to avoid discrepancy in deltaG(H(2)O) estimated from different models. PMID- 12547762 TI - Multiplexed-replica exchange molecular dynamics method for protein folding simulation. AB - Simulating protein folding thermodynamics starting purely from a protein sequence is a grand challenge of computational biology. Here, we present an algorithm to calculate a canonical distribution from molecular dynamics simulation of protein folding. This algorithm is based on the replica exchange method where the kinetic trapping problem is overcome by exchanging noninteracting replicas simulated at different temperatures. Our algorithm uses multiplexed-replicas with a number of independent molecular dynamics runs at each temperature. Exchanges of configurations between these multiplexed-replicas are also tried, rendering the algorithm applicable to large-scale distributed computing (i.e., highly heterogeneous parallel computers with processors having different computational power). We demonstrate the enhanced sampling of this algorithm by simulating the folding thermodynamics of a 23 amino acid miniprotein. We show that better convergence is achieved compared to constant temperature molecular dynamics simulation, with an efficient scaling to large number of computer processors. Indeed, this enhanced sampling results in (to our knowledge) the first example of a replica exchange algorithm that samples a folded structure starting from a completely unfolded state. PMID- 12547763 TI - Translocation of rodlike polymers through membrane channels. AB - A theory of channel-facilitated transport of long rodlike macromolecules through thin membranes under the influence of a driving force of arbitrary strength is developed. Analytic expressions are derived for the translocation probability and the Laplace transform of the probability density of time that a macromolecule spends in the channel. We also derive expressions for the (conditional) probability densities of time spent in the channel by translocating and nontranslocating (returning back) macromolecules. These results are used to study how the distribution of the macromolecule lifetime in the channel depends on a polymer chain length and the driving force. It is shown that depending on the values of the parameters, the lifetime probability density may have one or two peaks. Our theory is a generalization of the theory developed by Lubensky and Nelson, who were inspired by recent experiments on driven translocation of single stranded RNA and DNA molecules through single channels in narrow membranes. PMID- 12547764 TI - Analysis of metabolic capabilities using singular value decomposition of extreme pathway matrices. AB - It is now possible to construct genome-scale metabolic networks for particular microorganisms. Extreme pathway analysis is a useful method for analyzing the phenotypic capabilities of these networks. Many extreme pathways are needed to fully describe the functional capabilities of genome-scale metabolic networks, and therefore, a need exists to develop methods to study these large sets of extreme pathways. Singular value decomposition (SVD) of matrices of extreme pathways was used to develop a conceptual framework for the interpretation of large sets of extreme pathways and the steady-state flux solution space they define. The key results of this study were: 1), convex steady-state solution cones describing the potential functions of biochemical networks can be studied using the modes generated by SVD; 2), Helicobacter pylori has a more rigid metabolic network (i.e., a lower dimensional solution space and a more dominant first singular value) than Haemophilus influenzae for the production of amino acids; and 3), SVD allows for direct comparison of different solution cones resulting from the production of different amino acids. SVD was used to identify key network branch points that may identify key control points for regulation. Therefore, SVD of matrices of extreme pathways has proved to be a useful method for analyzing the steady-state solution space of genome-scale metabolic networks. PMID- 12547765 TI - MC-PHS: a Monte Carlo implementation of the primary hydration shell for protein folding and design. AB - A primary hydration shell (PHS) approach is developed for Monte Carlo simulations of conformationally rich macromolecular systems in an environment that efficiently captures principal solvation effects. It has been previously demonstrated that molecular dynamics using PHS is an efficient method to study peptide structure and dynamics in aqueous solution. Here, we extend the PHS approach to Monte Carlo simulations, whereby a stable shell of water molecules is maintained with a flexible, nonspherical, half-harmonic potential, tuned to maintain a constant restraining energy, with the difference between the restraint and shell energies used to dynamically adjust the shell radius. Examination of the shell and system size dependence of the restraining potential reveals its robustness. Moreover, its suitability for biomolecular simulations is evaluated using small spheres of water, hydration properties of small biological molecules, and configurational sampling of beta-hairpin pentapeptide YPGDV. This method, termed MC-PHS, appears to provide efficient representation of dominant solvation effects and should prove useful in the study of protein folding and design. PMID- 12547766 TI - Theoretical study of the structure and dynamic fluctuations of dioxolane-linked gramicidin channels. AB - Gramicidin is a hydrophobic peptide that assembles as a head-to-head dimer in lipid membranes to form water-filled channels selective to small monovalent cations. Two diastereoisomeric forms, respectively SS and RR, of chemically modified channels in which a dioxolane ring links the formylated N-termini of two gramicidin monomers, were shown to form ion channels. To investigate the structural basis underlying experimentally measured differences in proton conductance in the RR and SS channels, we construct atomic-resolution models of dioxolane-linked gramicidin dimers by analogy with the native dimer. A parametric description of the linker compatible with the CHARMM force field used for the peptide is derived by fitting geometry, vibrational frequencies, and energy to the results of ab initio calculations. The linker region of the modified gramicidin dimers is subjected to an extensive conformational search using high temperature simulated annealing, and free-energy surfaces underlying the structural fluctuations of the channel backbone at 298K are computed from molecular dynamics simulations. The overall secondary structure of the beta helical gramicidin pore is retained in both linked channels. The SS channel is found in a single conformation resembling that of the native dimer, with its peptide bonds undergoing rapid librations with respect to the channel axis. By contrast, its RR counterpart is characterized by local backbone distortions in which the two peptide bonds flanking the linker are markedly tilted in order to satisfy the pitch of the helix. In these distorted structures, each of the two carbonyl groups points either in or out of the lumen. Flipping these two peptides in and out involves thermally activated transitions, which results in four distinct conformational states at equilibrium with one another on a nanosecond time scale. This work opens the way to detailed comparative studies of structure function relationships in biological proton ducts. PMID- 12547767 TI - A mathematical model of the electrophysiological alterations in rat ventricular myocytes in type-I diabetes. AB - Our mathematical model of the rat ventricular myocyte (Pandit et al., 2001) was utilized to explore the ionic mechanism(s) that underlie the altered electrophysiological characteristics associated with the short-term model of streptozotocin-induced, type-I diabetes. The simulations show that the observed reductions in the Ca(2+)-independent transient outward K(+) current (I(t)) and the steady-state outward K(+) current (I(ss)), along with slowed inactivation of the L-type Ca(2+) current (I(CaL)), can result in the prolongation of the action potential duration, a well-known experimental finding. In addition, the model demonstrates that the slowed reactivation kinetics of I(t) in diabetic myocytes can account for the more pronounced rate-dependent action potential duration prolongation in diabetes, and that a decrease in the electrogenic Na(+)-K(+) pump current (I(NaK)) results in a small depolarization in the resting membrane potential (V(rest)). This depolarization reduces the availability of the Na(+) channels (I(Na)), thereby resulting in a slower upstroke (dV/dt(max)) of the diabetic action potential. Additional simulations suggest that a reduction in the magnitude of I(CaL), in combination with impaired sarcoplasmic reticulum uptake can lead to a decreased sarcoplasmic reticulum Ca(2+) load. These factors contribute to characteristic abnormal [Ca(2+)](i) homeostasis (reduced peak systolic value and rate of decay) in myocytes from diabetic animals. In combination, these simulation results provide novel information and integrative insights concerning plausible ionic mechanisms for the observed changes in cardiac repolarization and excitation-contraction coupling in rat ventricular myocytes in the setting of streptozotocin-induced, type-I diabetes. PMID- 12547769 TI - Protein interactions and membrane geometry. AB - The difficulty in growing crystals for x-ray diffraction analysis has hindered the determination of membrane protein structures. However, this is changing with the advent of a new method for growing high quality membrane protein crystals from the lipidic cubic phase. Although successful, the mechanism underlying this method has remained unclear. Here, we present a theoretical analysis of the process. We show that it is energetically favorable for proteins embedded in the highly curved cubic phase to cluster together in flattened regions of the membrane. This stabilizes the lamellar phase, permitting its outgrowth from the cubic phase. A kinetic barrier-crossing model is developed to determine the free energy barrier to crystallization from the time-dependent growth of protein clusters. Determining the values of key parameters provides both a rational basis for optimizing the experimental procedure for membrane proteins that have not yet been crystallized and insight into the analogous cubic to lamellar transitions in cells. We also discuss the implications of this mechanism for protein sorting at the exit sites of the Golgi and endoplasmic reticulum and the general stabilization of membrane structures. PMID- 12547768 TI - Regulation of protein mobility via thermal membrane undulations. AB - The in-plane diffusivelike motion of membrane bound proteins on the surface of cells is considered. We suggest, on the basis of theoretical arguments and simulation, that thermally excited undulations of the lipid bilayer may serve as a mechanism for proteins to hop between adjacent regions on the cell surface separated by barriers composed of internal cellular structure (e.g., the cytoskeleton). We specifically investigate the mobility of band 3 dimer on the surface of red blood cells where the spectrin cytoskeletal meshwork defines a series of "corrals" on the cell surface known to hinder protein motion. Previous models of this system have postulated that the cytoskeleton must deform to allow passage of membrane bound proteins out of these corral regions and have ignored fluctuations of the bilayer. Our model provides a complementary mechanism and we posit that the mobility of real proteins in real cells is likely the result of several mechanisms acting in parallel. PMID- 12547770 TI - Hybrid global optimization algorithms for protein structure prediction: alternating hybrids. AB - Hybrid global optimization methods attempt to combine the beneficial features of two or more algorithms, and can be powerful methods for solving challenging nonconvex optimization problems. In this paper, novel classes of hybrid global optimization methods, termed alternating hybrids, are introduced for application as a tool in treating the peptide and protein structure prediction problems. In particular, these new optimization methods take the form of hybrids between a deterministic global optimization algorithm, the alphaBB, and a stochastically based method, conformational space annealing (CSA). The alphaBB method, as a theoretically proven global optimization approach, exhibits consistency, as it guarantees convergence to the global minimum for twice-continuously differentiable constrained nonlinear programming problems, but can benefit from computationally related enhancements. On the other hand, the independent CSA algorithm is highly efficient, though the method lacks theoretical guarantees of convergence. Furthermore, both the alphaBB method and the CSA method are found to identify ensembles of low-energy conformers, an important feature for determining the true free energy minimum of the system. The proposed hybrid methods combine the desirable features of efficiency and consistency, thus enabling the accurate prediction of the structures of larger peptides. Computational studies for met enkephalin and melittin, employing sequential and parallel computing frameworks, demonstrate the promise for these proposed hybrid methods. PMID- 12547771 TI - Long-range signal transmission in autocrine relays. AB - Intracellular signaling induced by peptide growth factors can stimulate secretion of these molecules into the extracellular medium. In autocrine and paracrine networks, this can establish a positive feedback loop between ligand binding and ligand release. When coupled to intercellular communication by autocrine ligands, this positive feedback can generate constant-speed traveling waves. To demonstrate that, we propose a mechanistic model of autocrine relay systems. The model is relevant to the physiology of epithelial layers and to a number of in vitro experimental formats. Using asymptotic and numerical tools, we find that traveling waves in autocrine relays exist and have a number of unusual properties, such as an optimal ligand binding strength necessary for the maximal speed of propagation. We compare our results to recent observations of autocrine and paracrine systems and discuss the steps toward experimental tests of our predictions. PMID- 12547772 TI - Ising model of cardiac thin filament activation with nearest-neighbor cooperative interactions. AB - We have developed a model of cardiac thin filament activation using an Ising model approach from equilibrium statistical physics. This model explicitly represents nearest-neighbor interactions between 26 troponin/tropomyosin units along a one-dimensional array that represents the cardiac thin filament. With transition rates chosen to match experimental data, the results show that the resulting force-pCa (F-pCa) relations are similar to Hill functions with asymmetries, as seen in experimental data. Specifically, Hill plots showing (log(F/(1-F)) vs. log [Ca]) reveal a steeper slope below the half activation point (Ca(50)) compared with above. Parameter variation studies show interplay of parameters that affect the apparent cooperativity and asymmetry in the F-pCa relations. The model also predicts that Ca binding is uncooperative for low [Ca], becomes steeper near Ca(50), and becomes uncooperative again at higher [Ca]. The steepness near Ca(50) mirrors the steep F-pCa as a result of thermodynamic considerations. The model also predicts that the correlation between troponin/tropomyosin units along the one-dimensional array quickly decays at high and low [Ca], but near Ca(50), high correlation occurs across the whole array. This work provides a simple model that can account for the steepness and shape of F-pCa relations that other models fail to reproduce. PMID- 12547773 TI - Permeant cations and blockers modulate pH gating of ROMK channels. AB - External potassium (K) activates the inward rectifier ROMK (K(ir)1.1) by altering the pH gating of the channel. The present study examines this link between external K and internal pH sensitivity using both the two-electrode voltage clamp and the perfused, cut-open Xenopus oocyte preparation. Elevating extracellular K from 1 mM to 10 mM to 100 mM activated ROMK channels by shifting their apparent pK(a) from 7.2 +/- 0.1 (n = 6) in 1 mM K, to 6.9 +/- 0.02 (n = 5) in 10 mM K, and to 6.6 +/- 0.03 (n = 5) in 100 mM K. At any given internal pH, the number of active ROMK channels is a saturating function of external [K]. Extracellular Cs (which blocks almost all inward K current) also stimulated outward ROMK conductance (at constant 1 mM external K) by shifting the apparent pK(a) of ROMK from 7.2 +/- 0.1 (n = 6) in 1 mM K to 6.8 +/- 0.01 (n = 4) in 1 mM K + 104 mM Cs. Surprisingly, the binding and washout of the specific blocker, Tertiapin-Q, also activated ROMK in 1 mM K and caused a comparable shift in apparent pK(a). These results are interpreted in terms of both a three-state kinetic model and a two gate structural model that is based on results with KcsA in which the selectivity filter can assume either a high or low K conformation. In this context, external K, Cs, and Tertiapin-Q activate ROMK by destabilizing the low-K (collapsed) configuration of the selectivity filter. PMID- 12547774 TI - MIC channels are inhibited by internal divalent cations but not ATP. AB - TRPM7 channels are nonselective cation channels that possess a functional alpha kinase domain. It has been proposed that heterologously expressed TRPM7 channels are activated (Runnels et al., 2001) or inhibited (Nadler et al., 2001) by dialyzing the cell with millimolar levels of ATP. The endogenous correlate of TRPM7 has been identified in T-lymphocytes and RBL (rat basophilic leukemia) cells and named MagNuM (for Mg(2+)-nucleotide-inhibited metal) or MIC (for Mg(2+) inhibited cation). Here, we report that internal Mg(2+) rather than MgATP inhibits this current. Cytoplasmic MgATP, supplied by dialysis at millimolar concentrations, effectively inhibits only when a weak Mg(2+) chelator is present in the pipette solution. Thus, MgATP acts as a source of Mg(2+) rather than a source of ATP. Using an externally accessible site within the pore of the MIC channel itself as a bioassay, we show that equimolar MgCl(2) and MgATP solutions contain similar amounts of free Mg(2+), explaining the fact that numeric values of Mg(2+) and MgATP concentrations necessary for complete inhibition are the same. Furthermore, we demonstrate that Mg(2+) is not unique in its inhibitory action, as Ba(2+), Sr(2+), Zn(2+), and Mn(2+) can substitute for Mg(2+), causing complete inhibition. We conclude that MIC current inhibition occurs simply by divalent cations. PMID- 12547775 TI - The link between ion permeation and inactivation gating of Kv4 potassium channels. AB - Kv4 potassium channels undergo rapid inactivation but do not seem to exhibit the classical N-type and C-type mechanisms present in other Kv channels. We have previously hypothesized that Kv4 channels preferentially inactivate from the preopen closed state, which involves regions of the channel that contribute to the internal vestibule of the pore. To further test this hypothesis, we have examined the effects of permeant ions on gating of three Kv4 channels (Kv4.1, Kv4.2, and Kv4.3) expressed in Xenopus oocytes. Rb(+) is an excellent tool for this purpose because its prolonged residency time in the pore delays K(+) channel closing. The data showed that, only when Rb(+) carried the current, both channel closing and the development of macroscopic inactivation are slowed (1.5- to 4 fold, relative to the K(+) current). Furthermore, macroscopic Rb(+) currents were larger than K(+) currents (1.2- to 3-fold) as the result of a more stable open state, which increases the maximum open probability. These results demonstrate that pore occupancy can influence inactivation gating in a manner that depends on how channel closing impacts inactivation from the preopen closed state. By examining possible changes in ionic selectivity and the influence of elevating the external K(+) concentration, additional experiments did not support the presence of C-type inactivation in Kv4 channels. PMID- 12547776 TI - Ca2+ current and charge movements in skeletal myotubes promoted by the beta subunit of the dihydropyridine receptor in the absence of ryanodine receptor type 1. AB - The beta-subunit of the dihydropyridine receptor (DHPR) enhances the Ca(2+) channel and voltage-sensing functions of the DHPR. In skeletal myotubes, there is additional modulation of DHPR functions imposed by the presence of ryanodine receptor type-1 (RyR1). Here, we examined the participation of the beta-subunit in the expression of L-type Ca(2+) current and charge movements in RyR1 knock-out (KO), beta1 KO, and double beta1/RyR1 KO myotubes generated by mating heterozygous beta1 KO and RyR1 KO mice. Primary myotube cultures of each genotype were transfected with various beta-isoforms and then whole-cell voltage-clamped for measurements of Ca(2+) and gating currents. Overexpression of the endogenous skeletal beta1a isoform resulted in a low-density Ca(2+) current either in RyR1 KO (36 +/- 9 pS/pF) or in beta1/RyR1 KO (34 +/- 7 pS/pF) myotubes. However, the heterologous beta2a variant with a double cysteine motif in the N-terminus (C3, C4), recovered a Ca(2+) current that was entirely wild-type in density in RyR1 KO (195 +/- 16 pS/pF) and was significantly enhanced in double beta1/RyR1 KO (115 +/ 18 pS/pF) myotubes. Other variants tested from the four beta gene families (beta1a, beta1b, beta1c, beta3, and beta4) were unable to enhance Ca(2+) current expression in RyR1 KO myotubes. In contrast, intramembrane charge movements in beta2a-expressing beta1a/RyR1 KO myotubes were significantly lower than in beta1a expressing beta1a/RyR1 KO myotubes, and the same tendency was observed in the RyR1 KO myotube. Thus, beta2a had a preferential ability to recover Ca(2+) current, whereas beta1a had a preferential ability to rescue charge movements. Elimination of the double cysteine motif (beta2a C3,4S) eliminated the RyR1 independent Ca(2+) current expression. Furthermore, Ca(2+) current enhancement was observed with a beta2a variant lacking the double cysteine motif and fused to the surface membrane glycoprotein CD8. Thus, tethering the beta2a variant to the myotube surface activated the DHPR Ca(2+) current and bypassed the requirement for RyR1. The data suggest that the Ca(2+) current expressed by the native skeletal DHPR complex has an inherently low density due to inhibitory interactions within the DHPR and that the beta1a-subunit is critically involved in process. PMID- 12547777 TI - Methionine changes in bacteriorhodopsin detected by FTIR and cell-free selenomethionine substitution. AB - Bacteriorhodopsin (BR) is an integral membrane protein, which functions as a light-driven proton pump in Halobacterium salinarum. We report evidence that one or more methionine residues undergo a structural change during the BR-->M portion of the BR photocycle. Selenomethionine was incorporated into BR using a cell-free protein translation system containing an amino acid mixture with selenomethionine substituted for methionine. BR-->M FTIR difference spectra recorded for unlabeled and selenomethionine-labeled cell-free expressed BR closely resemble the spectra of in vivo expressed BR. However, reproducible changes occur in two regions near 1,284 and 900 cm(-1) due to selenomethionine incorporation. Isotope labeled tyrosine was also co-incorporated with selenomethionine in order to confirm these assignments. Based on recent x-ray crystallographic studies, likely methionines which give rise to the FTIR difference bands are Met-118 and Met-145, which are located inside the retinal binding pocket and in a position to constrain the motion of retinal during photoisomerization. The assignment of methionine bands in the FTIR difference spectrum of BR provides a means to study methionine chromophore interaction under physiological conditions. More generally, combining cell-free incorporations of selenomethionine into proteins with FTIR difference spectroscopy provides a useful method for investigating the role of methionines in protein structure and function. PMID- 12547778 TI - Highly accurate classification of Watson-Crick basepairs on termini of single DNA molecules. AB - We introduce a computational method for classification of individual DNA molecules measured by an alpha-hemolysin channel detector. We show classification with better than 99% accuracy for DNA hairpin molecules that differ only in their terminal Watson-Crick basepairs. Signal classification was done in silico to establish performance metrics (i.e., where train and test data were of known type, via single-species data files). It was then performed in solution to assay real mixtures of DNA hairpins. Hidden Markov Models (HMMs) were used with Expectation/Maximization for denoising and for associating a feature vector with the ionic current blockade of the DNA molecule. Support Vector Machines (SVMs) were used as discriminators, and were the focus of off-line training. A multiclass SVM architecture was designed to place less discriminatory load on weaker discriminators, and novel SVM kernels were used to boost discrimination strength. The tuning on HMMs and SVMs enabled biophysical analysis of the captured molecule states and state transitions; structure revealed in the biophysical analysis was used for better feature selection. PMID- 12547779 TI - Regulation of the fast vacuolar channel by cytosolic and vacuolar potassium. AB - At resting cytosolic Ca(2)(+), passive K(+) conductance of a higher plant tonoplast is likely dominated by fast vacuolar (FV) channels. This patch-clamp study describes K(+)-sensing behavior of FV channels in Beta vulgaris taproot vacuoles. Variation of K(+) between 10 and 400 mM had little effect on the FV channel conductance, but a pronounced one on the open probability. Shift of the voltage dependence by cytosolic K(+) could be explained by screening of the negative surface charge with a density sigma = 0.25 e(-)/nm(2). Vacuolar K(+) had a specific effect on the FV channel gating at negative potentials without significant effect on closed-open transitions at positive ones. Due to K(+) effects at either membrane side, the potential at which the FV channel has minimal activity was always situated at approximately 50 mV below the potassium equilibrium potential, E(K(+)). At tonoplast potentials below or equal to E(K(+)), the FV channel open probability was almost independent on the cytosolic K(+) but varied in a proportion to the vacuolar K(+). Therefore, the release of K(+) from the vacuole via FV channels could be controlled by the vacuolar K(+) in a feedback manner; the more K(+) is lost the lower will be the transport rate. PMID- 12547781 TI - Material studies of lipid vesicles in the L(alpha) and L(alpha)-gel coexistence regimes. AB - In this work, we utilize micropipette aspiration and fluorescence imaging to examine the material properties of lipid vesicles made from mixtures of palmitoyloleoylphosphocholine (POPC) and dipalmitoylphosphatidylcholine (DPPC). At elevated temperatures/low DPPC fractions, these lipids are in a miscible liquid crystalline (L(alpha)) state, whereas at lower temperatures/higher DPPC fractions they phase-separate into L(alpha) and gel phases. We show that the elastic modulus, K, and critical tension, tau(c), of L(alpha) vesicles are independent of DPPC fraction. However, as the sample temperature is increased from 15 degrees C to 45 degrees C, we measure decreases in both K and tau(c) of 20% and 50%, respectively. The elasticity change is likely driven by a change in interfacial tension. We describe the reduction in critical tension using a simple model of thermally activated membrane pores. Vesicles with two-phase coexistence exhibit material properties that differ from L(alpha) vesicles including critical tensions that are 20-40% lower. Fluorescence imaging of phase coexistent POPC/DPPC vesicles shows that the DPPC-rich domains exist in an extended network structure that exhibits characteristics of a solid. This gel network explains many of the unusual material properties of two-phase membranes. PMID- 12547780 TI - Properties of palmitoyl phosphatidylcholine, sphingomyelin, and dihydrosphingomyelin bilayer membranes as reported by different fluorescent reporter molecules. AB - The properties of vesicle membranes prepared from 16:0-SM, 16:0-DHSM, or DPPC were characterized using steady-state and time-resolved fluorescence spectroscopy and different fluorescent reporter molecules. The acyl-chain region was probed using free and phospholipid-bound 1,6-diphenyl-1,3,5-hexatriene. 16:0-DHSM was found to be the more ordered than both DPPC and 16:0-SM 5 degrees C below and above melting temperature. Interfacial properties of the phospholipid bilayers were examined using 6-dodecanoyl-2-dimethyl-aminonaphthalene (Laurdan), 6 propionyl-2-dimethyl-amino-naphthalene (Prodan), and dansyl-PE. Laurdan and Prodan reported that the two sphingomyelin (SM) membrane interfaces were clearly different from the DPPC membrane interface, whereas the two SM membrane interfaces had more similar properties (both in gel and liquid-crystalline phase). Prodan partition studies showed that membrane resistance to Prodan partitioning increased in the order: 16:0-SM < DPPC < 16:0-DHSM. The degree to which dansyl-PE is exposed to water reflects the structural properties of the membrane-water interface. By comparing the lifetime of dansyl-PE in water and deuterium oxide solution, we could show that the degree to which the dansyl moiety was exposed to water in the membranes increased in the order: 16:0-SM < DPPC < 16:0-DHSM. In conclusion, this study has shown that DHSM forms more ordered bilayers than acyl-chain matched SM or phosphatidylcholine, even in the liquid-crystalline state. PMID- 12547782 TI - Hydration-driven transport of deformable lipid vesicles through fine pores and the skin barrier. AB - We studied aggregate transport through semipermeable, nano-porous barriers experimentally and theoretically. By measuring and modeling the effect of hydration gradient across such barriers, spontaneous transbarrier transport of suitable lipid aggregates in vesicular form was proven to be driven by partial aggregate dehydration at the application site. By generalizing the Onsager transport model we derived a set of equations that rationalize all pertinent observations. Dehydration-induced vesicle motion starts with a lag time. This corresponds to the time needed to reach the limiting vesicle hydration; both are proportional to the starting excess water volume and decrease with increasing relative humidity at application site. The rate of transbarrier transport is insensitive to these parameters but increases with vesicle deformability and volume exchange capability. Both these properties depend on membrane composition. Reversible demixing of bilayer components is the cause of nonlinear bilayer characteristics and also potentially affects the effective membrane hydrophilicity. High hydrophilicity of vesicle surface and extreme aggregate shape adaptability together are necessary for successful material transport across the skin. This demonstrates the significance of basic biophysical investigations for better understanding of biological systems and for the practical use of artificial, nature-inspired carriers in drug delivery. PMID- 12547783 TI - Intramembrane polarity by electron spin echo spectroscopy of labeled lipids. AB - The association of water (D(2)O) with phospholipid membranes was studied by using pulsed-electron spin resonance techniques. We measured the deuterium electron spin echo modulation of spin-labeled phospholipids by D(2)O in membranes of dipalmitoyl phosphatidylcholine with and without 50 mol% of cholesterol. The Fourier transform of the relaxation-corrected two-pulse echo decay curve reveals peaks, at one and two times the deuterium NMR frequency, that arise from the dipolar hyperfine interaction of the deuterium nucleus with the unpaired electron spin of the nitroxide-labeled lipid. For phosphatidylcholine spin-labeled at different positions down the sn-2 chain, the amplitude of the deuterium signal decreases toward the center of the membrane, and is reduced to zero from the C-12 atom position onward. At chain positions C-5 and C-7 closer to the phospholipid headgroups, the amplitude of the deuterium signal is greater in the presence of cholesterol than in its absence. These results are in good agreement with more indirect measurements of the transmembrane polarity profile that are based on the (14)N-hyperfine splittings in the conventional continuous-wave electron spin resonance spectrum. PMID- 12547784 TI - Structural proton diffusion along lipid bilayers. AB - For H(+) transport between protein pumps, lateral diffusion along membrane surfaces represents the most efficient pathway. Along lipid bilayers, we measured a diffusion coefficient of 5.8 x 10(-5) cm(2) s(-1). It is too large to be accounted for by vehicle diffusion, considering proton transport by acid carriers. Such a speed of migration is accomplished only by the Grotthuss mechanism involving the chemical exchange of hydrogen nuclei between hydrogen bonded water molecules on the membrane surface, and the subsequent reorganization of the hydrogen-bonded network. Reconstitution of H(+)-binding sites on the membrane surface decreased the velocity of H(+) diffusion. In the absence of immobile buffers, structural (Grotthuss) diffusion occurred over a distance of 100 micro m as shown by microelectrode aided measurements of the spatial proton distribution in the immediate membrane vicinity and spatially resolved fluorescence measurements of interfacial pH. The efficiency of the anomalously fast lateral diffusion decreased gradually with an increase in mobile buffer concentration suggesting that structural diffusion is physiologically important for distances of approximately 10 nm. PMID- 12547785 TI - Effects of natural and enantiomeric cholesterol on the thermotropic phase behavior and structure of egg sphingomyelin bilayer membranes. AB - Phospholipids, sphingolipids, and sterols are the major lipid components of the plasma membranes of eukaryotic cells. Because these three lipid classes occur naturally as enantiomerically pure compounds, enantiospecific lipid-lipid and lipid-sterol interactions could in principle occur in the lipid bilayers of eukaryotic plasma membranes. Although previous biophysical studies of phospholipid and phospholipid-sterol model membrane systems have consistently failed to observe such enantiomerically selective interactions, a recent monolayer study of the interactions of natural and enantiomeric cholesterol with egg sphingomyelin has apparently revealed the existence of enantiospecific sterol sphingolipid interactions. To determine whether enantiospecific sterol sphingolipid interactions also occur in more biologically relevant lipid-bilayer systems, differential scanning calorimetric, x-ray diffraction, and neutral buoyant-density measurements were utilized to study the effects of natural and enantiomeric cholesterol on the thermotropic phase behavior and structure of egg sphingomyelin bilayers. The calorimetry experiments show that the natural and enantiomeric cholesterol have essentially identical effects on the temperature, enthalpy, and cooperativity of the gel/liquid-crystalline phase transition of egg sphingomyelin bilayers within the limits of experimental error. As well, the x ray diffraction and neutral buoyancy experiments indicate that bilayers formed from mixtures of natural or enantiomeric cholesterol and egg sphingomyelin have, within experimental uncertainty, the same structure and mass density. We thus conclude that significant enantioselective cholesterol-sphingolipid interactions do not occur in this lipid-bilayer model membrane system. PMID- 12547786 TI - Chemical decoupling of ATPase activation and force production from the contractile cycle in myosin by steric hindrance of lever-arm movement. AB - The myosin motor protein generates force in muscle by hydrolyzing Adenosine 5' triphosphate (ATP) while interacting transiently with actin. Structural evidence suggests the myosin globular head (subfragment 1 or S1) is articulated with semi rigid catalytic and lever-arm domains joined by a flexible converter domain. According to the prevailing hypothesis for energy transduction, ATP binding and hydrolysis in the catalytic domain drives the relative movement of the lever arm. Actin binding and reversal of the lever-arm movement (power stroke) applies force to actin. These domains interface at the reactive lysine, Lys84, where trinitrophenylation (TNP-Lys84-S1) was observed in this work to block actin activation of myosin ATPase and in vitro sliding of actin over myosin. TNP-Lys84 S1's properties and interactions with actin were examined to determine how trinitrophenylation causes these effects. Weak and strong actin binding, the rate of mantADP release from actomyosin, and actomyosin dissociation by ATP were equivalent in TNP-Lys84-S1 and native S1. Molecular dynamics calculations indicate that lever-arm movement inhibition during ATP hydrolysis and the power stroke is caused by steric clashes between TNP and the converter or lever-arm domains. Together these findings suggest that TNP uncouples actin activation of myosin ATPase and the power stroke from other steps in the contraction cycle by inhibiting the converter and lever-arm domain movements. PMID- 12547787 TI - The calcium-saturated cTnI/cTnC complex: structure of the inhibitory region of cTnI. AB - The contiguous inhibitory and regulatory regions of troponin I in the heterotrimeric troponin complex play a critical role in Ca(2+) activation of striated muscle. Knowledge of the structure of this critical region within the complex will enhance efforts toward understanding regulatory mechanisms. Toward this goal, we have used simulated annealing to study the structure of the inhibitory and regulatory regions of cardiac muscle troponin I in the calcium saturated complex formed between cardiac troponin C and cardiac troponin I. We have incorporated distances determined experimentally by Forster resonance energy transfer in the full-length complex, rather than using peptides derived from cTnI. For these models, we assume a helix-loop-helix conformation for the inhibitory region. We have found several structures that satisfy the experimental constraints fairly well. Although it is not possible to eliminate any of these models at this time, future studies with additional experimental restraints will yield insights on the mechanisms of calcium regulation in cardiac muscle. PMID- 12547788 TI - Voltage-activated calcium signals in myotubes loaded with high concentrations of EGTA. AB - In the present study we describe the analysis of optically recorded whole cell Ca(2+) transients elicited by depolarization in cultured skeletal myotubes. Myotubes were obtained from the mouse muscle-derived cell line C2C12 and from mouse satellite cells. The cells were voltage-clamped and perfused with an artificial intracellular solution containing 15 mM EGTA to ensure that the bulk of the Ca(2+) mobilized by depolarization is bound to this extrinsic buffer. The apparent on- and off-rate constants of EGTA and the dissociation rate constant of fura-2 in the cell were estimated by investigating the Ca(2+)-dependence of kinetic components of the fluorescence decay after repolarization. These parameters were used to calculate the time course of the total voltage-controlled flux of Ca(2+) to the myoplasmic space (Ca(2+) input flux). The validity of the procedure was confirmed by model simulations using artificial Ca(2+) input fluxes. Both C2C12 and primary-cultured myotubes showed a very similar phasic tonic time course of the Ca(2+) input flux. In most measurements, the input flux was considerably larger and showed a different time course than the estimated Ca(2+) flux carried by the L-type Ca(2+) channels, indicating that it consists mainly of voltage-controlled Ca(2+) release from the sarcoplasmic reticulum. In cells with extremely small fluorescence transients, the calculated input fluxes matched the kinetic characteristics of the Ca(2+) inward current, indicating that Ca(2+) release was absent. These measurements served as a control for the fidelity of the fluorimetric flux analysis. The procedures promise a deeper insight into alterations of Ca(2+) release gating in studies employing myotube expression systems for mutant or chimeric protein components of excitation contraction coupling. PMID- 12547789 TI - Location of ryanodine and dihydropyridine receptors in frog myocardium. AB - Frog myocardium depends almost entirely on calcium entry from extracellular spaces for its beat-to-beat activation. Atrial myocardium additionally shows internal calcium release under certain conditions, but internal release in the ventricle is absent or very low. We have examined the content and distribution of the sarcoplasmic reticulum (SR) calcium release channels (ryanodine receptors, RyRs) and the surface membrane calcium channels (dihydropyridine receptors, DHPRs) in myocardium from the two atria and the ventricle of the frog heart using binding of radioactive ryanodine, immunolabeling of RyR and DHPR, and thin section and freeze-fracture electron microscopy. In cells from both types of chambers, the SR forms peripheral couplings and in both chambers peripheral couplings colocalize with clusters of DHPRs. However, although a low level of high affinity binding of ryanodine is detectable and RyRs are present in peripheral couplings of the atrium, the ventricle shows essentially no ryanodine binding and RyRs are not detectable either by electron microscopy or immunolabeling. The results are consistent with the lack of internal calcium release in the ventricle, and raise questions regarding the significance of DHPR at peripheral couplings in the absence of RyR. Interestingly, the free SR membrane in both heart chambers shows a low but equal density of intramembrane particles representing the Ca(2+) ATPase. PMID- 12547790 TI - An x-ray diffraction study on early structural changes in skeletal muscle contraction. AB - Structural changes in frog skeletal muscle were studied using x-ray diffraction with a time resolution of 0.53-1.02 ms after a single electrical stimulus at 8 degrees C. Tension began to drop at 6 ms (latency relaxation), reached a minimum at 8 ms, and then twitch tension developed. The intensity of the meridional reflection at 1/38.5 nm(-1), from troponin molecules on the thin filament, began to increase at 4-5 ms and reached a maximum at approximately 12 ms. The meridional reflections based on the myosin 43-nm repeat began to decrease when the tension began to develop. The peak position of the third-order myosin meridional reflection began to shift toward the higher angle at approximately 5 ms, reached a maximum shift (0.02%) at 10 ms, and then moved toward the lower angle. The intensity of the second actin layer line at 1/18 nm(-1) in the axial direction, which was measured at 12 degrees C, began to rise at 5 ms, whereas the latency relaxation started at 3.5 ms. These results suggest that 1), the Ca(2+) induced structural changes in the thin filament and a structural change in the thick filament have already taken place during latency relaxation; and 2), the Ca(2+) regulation of the thin filament is highly cooperative. PMID- 12547791 TI - Equilibrium and transition between single- and double-headed binding of kinesin as revealed by single-molecule mechanics. AB - Kinesin is a processive motor protein that "walks" on a microtubule toward its plus end. We reported previously that the distribution of unbinding force and elastic modulus for a single kinesin-microtubule complex was either unimodal or bimodal depending on the nucleotide states of the kinesin heads, hence showing that the kinesin may bind the microtubule either with one head or with both heads at once. Here, we found that the shape of the unbinding-force distribution depends both on the loading rate and on the manner of loading not only in the presence of AMP-PNP but also in the absence of nucleotides. Irrespective of the nucleotide state and the loading conditions examined here, the unbinding force obtained by loading directed toward the minus end of microtubule was 45% greater than that for plus end-directed loading. These results could be explained by a model in which equilibrium exists between single- and double-headed binding and the load (F) dependence of lifetime, tau(F), of each binding is expressed by tau(F) = tau(0)exp(-Fd/k(B)T), where tau(0) is the lifetime without external load and d a characteristic distance, both of which depend on single- or double-headed binding, k(B), the Boltzmann constant and T, the absolute temperature. The model analysis showed that the forward and backward rates of transition from single- to double-headed binding are 2 and 0.2/s for the AMP-PNP state, and 70 and 7/s for the nucleotide-free state. Moreover, in the presence of AMP-PNP, we detected the moment of transition from single- to double-headed binding through an abrupt increase in the elastic modulus and estimated the transition rate to be approximately 1/s, which is consistent with the model analysis. PMID- 12547792 TI - The structure of DNA within cationic lipid/DNA complexes. AB - The structure of DNA within CLDCs used for gene delivery is controversial. Previous studies using CD have been interpreted to indicate that the DNA is converted from normal B to C form in complexes. This investigation reexamines this interpretation using CD of model complexes, FTIR as well as Raman spectroscopy and molecular dynamics simulations to address this issue. CD spectra of supercoiled plasmid DNA undergo a significant loss of rotational strength in the signal near 275 nm upon interaction with either the cationic lipid dimethyldioctadecylammonium bromide or 1,2-dioleoyltrimethylammonium propane. This loss of rotational strength is shown, however, by both FTIR and Raman spectroscopy to occur within the parameters of the B-type conformation. Contributions of absorption flattening and differential scattering to the CD spectra of complexes are unable to account for the observed spectra. Model studies of the CD of complexes prepared from synthetic oligonucleotides of varying length suggest that significant reductions in rotational strength can occur within short stretches of DNA. Furthermore, some alteration in the hydrogen bonding of bases within CLDCs is indicated in the FTIR and Raman spectroscopy results. In addition, alterations in base stacking interactions as well as hydrogen bonding are suggested by molecular dynamics simulations. A global interpretation of all of the data suggests the DNA component of CLDCs remains in a variant B form in which base/base interactions are perturbed. PMID- 12547793 TI - The role of a microscopic colloidally stabilized phase in solubilizing oligoamine condensed DNA complexes. AB - DNA complexes of spermine and spermidine become resolubilized at very high concentrations of the oligoamine. It has been postulated that high oligoamine concentrations shift the DNA from the globule back to the coil phase. The present study indicates that DNA resolubilization at high concentrations of spermine and spermidine is explained by formation of small particles of condensed DNA that cannot be precipitated by centrifugation. The fact that DNA stays condensed during resolubilization was confirmed using a relatively new condensation assay and three independent microscopic techniques. A considerable portion of DNA was found to be in particles with diameter <100 nm. Formation of such small particles is likely to be caused by colloidal forces. The ability to form small, condensed DNA particles in solutions that contain high concentrations of oligocation should aid in the design of synthetic DNA vectors for gene transfer and gene therapy and in the handling of DNA for diagnostic studies. PMID- 12547794 TI - Fluorescence resonance energy transfer over approximately 130 basepairs in hyperstable lac repressor-DNA loops. AB - Lac repressor (LacI) binds two operator DNA sites, looping the intervening DNA. DNA molecules containing two lac operators bracketing a sequence-directed bend were previously shown to form hyperstable LacI-looped complexes. Biochemical studies suggested that orienting the operators outward relative to the bend direction (in construct 9C14) stabilizes a positively supercoiled closed form, with a V-shaped LacI, but that the most stable loop construct (11C12) is a more open form. Here, fluorescence resonance energy transfer (FRET) is measured on DNA loops, between fluorescein and TAMRA attached near the two operators, approximately 130 basepairs apart. For 9C14, efficient LacI-induced energy transfer ( approximately 74% based on donor quenching) confirms that the designed DNA shape can force the looped complex into a closed form. From enhanced acceptor emission, correcting for observed donor-dependent quenching of acceptor fluorescence, approximately 52% transfer was observed. Time-resolved FRET suggests that this complex exists in both closed- and open form populations. Less efficient transfer, approximately 10%, was detected for DNA-LacI sandwiches and 11C12-LacI, consistent with an open form loop. This demonstration of long-range FRET in large DNA loops confirms that appropriate DNA design can control loop geometry. LacI flexibility may allow it to maintain looping with other proteins bound or under different intracellular conditions. PMID- 12547795 TI - Self-regulation phenomena applied to bacterial reaction centers: 2. Nonequilibrium adiabatic potential: dark and light conformations revisited. AB - Experimental and theoretical results in support of nonlinear dynamic behavior of photosynthetic reaction centers under light-activated conditions are presented. Different conditions of light adaptation allow for preparation of reaction centers in either of two different conformational states. These states were detected both by short actinic flashes and by the switching of the actinic illumination level between different stationary state values. In the second method, the equilibration kinetics of reaction centers isolated from Rhodobacter sphaeroides were shown to be inherently biphasic. The fast and slow equilibration kinetics are shown to correspond to electron transfer (charge separation) at a fixed structure and to combined electron-conformational transitions governed by the bounded diffusion along the potential surface, respectively. The primary donor recovery kinetics after an actinic flash revealed a pronounced dependence on the time interval (deltat) between cessation of a lengthy preillumination of a sample and the actinic flash. A pronounced slow relaxation component with a decay half time of more than 50 s was measured for deltat > 10 s. This component corresponds to charge recombination in reaction centers for which light-induced structural changes have not relaxed completely before the flash. The amplitude of this component depended on the conditions of the sample preparation, specifically on the type of detergent used in the preparation. The redox potential parameters as well as the structural diffusion constants were estimated for samples prepared in different ways. PMID- 12547796 TI - Excitation energy transfer dynamics and excited-state structure in chlorosomes of Chlorobium phaeobacteroides. AB - The excited-state relaxation within bacteriochlorophyll (BChl) e and a in chlorosomes of Chlorobium phaeobacteroides has been studied by femtosecond transient absorption spectroscopy at room temperature. Singlet-singlet annihilation was observed to strongly influence both the isotropic and anisotropic decays. Pump intensities in the order of 10(11) photons x pulse(-1) x cm(-2) were required to obtain annihilation-free conditions. The most important consequence of applied very low excitation doses is an observation of a subpicosecond process within the BChl e manifold (approximately 200-500 fs), manifesting itself as a rise in the red part of the Q(y) absorption band of the BChl e aggregates. The subsequent decay of the kinetics measured in the BChl e region and the corresponding rise in the baseplate BChl a is not single exponential, and at least two components are necessary to fit the data, corresponding to several BChl e-->BChl a transfer steps. Under annihilation-free conditions, the anisotropic kinetics show a generally slow decay within the BChl e band (10-20 ps) whereas it decays more rapidly in the BChl a region ( approximately 1 ps). Analysis of the experimental data gives a detailed picture of the overall time evolution of the energy relaxation and energy transfer processes within the chlorosome. The results are interpreted within an exciton model based on the proposed structure. PMID- 12547797 TI - Deuterium isotope effects in the photocycle transitions of the photoactive yellow protein. AB - The Photoactive Yellow Protein (PYP) from Halorhodospira halophila (formerly Ectothiorhodospira halophila) is increasingly used as a model system. As such, a thorough understanding of the photocycle of PYP is essential. In this study we have combined information from pOH- (or pH-) dependence and (kinetic) deuterium isotope effects to elaborate on existing photocycle models. For several characteristics of PYP we were able to make a distinction between pH- and pOH dependence, a nontrivial distinction when comparing data from samples dissolved in H(2)O and D(2)O. It turns out that most characteristics of PYP are pOH dependent. We confirmed the existence of a pB' intermediate in the pR to pB transition of the photocycle. In addition, we were able to show that the pR to pB' transition is reversible, which explains the previously observed biexponential character of the pR-to-pB photocycle step. Also, the absorption spectrum of pB' is slightly red-shifted with respect to pB. The recovery of the pG state is accompanied by an inverse kinetic deuterium isotope effect. Our interpretation of this is that before the chromophore can be isomerized, it is deprotonated by a hydroxide ion from solution. From this we propose a new photocycle intermediate, pB(deprot), from which pG is recovered and which is in equilibrium with pB. This is supported in our data through the combination of the observed pOH and pH dependence, together with the kinetic deuterium isotope effect. PMID- 12547798 TI - Phot-LOV1: photocycle of a blue-light receptor domain from the green alga Chlamydomonas reinhardtii. AB - The "Phot" protein family comprises blue-light photoreceptors that consist of two flavin mononucleotide (FMN)-binding LOV (light, oxygen, and voltage) domains and a serine/threonine kinase domain. We have investigated the LOV1 domain of Phot1 from Chlamydomonas reinhardtii by time-resolved absorption spectroscopy. Photoexcitation of the dark form, LOV1-447, causes transient bleaching and formation of two spectrally similar red-shifted intermediates that are both assigned to triplet states of the FMN. The triplet states decay with time constants of 800 ns and 4 micro s with an efficiency of >90% into a blue-shifted intermediate, LOV1-390, that is attributed to a thiol adduct of cysteine 57 to FMN C(4a). LOV1-390 reverts to the dark form in hundreds of seconds, the time constant being dependent on pH and salt concentration. In the mutant C57S, where the thiol adduct cannot be formed, the triplet state displays an oxygen-dependent decay directly to the dark form. We present here a spectroscopic characterization of an algal sensory photoreceptor in general and of a LOV1 domain photocycle in particular. The results are discussed with respect to the behavior of the homologous LOV2 domain from oat. PMID- 12547799 TI - Characterization of the photochemical reaction cycle of proteorhodopsin. AB - Absorption changes in the photocycle of the recently described retinal protein, proteorhodopsin, are analyzed. The transient spectra at pH 9.5, where it acts as a light-driven proton pump, reveal the existence of three spectrally different intermediates, K, M, and N, named in analogy with the photointermediates of bacteriorhodopsin. Model analysis based on time-dependent absorption kinetic signals at four wavelengths suggested the existence of two more spectrally silent intermediates and lead to a sequential reaction scheme with five intermediates, K, M(1), M(2), N, and PR', before decay to the initial state PR. An L-like intermediate was not observed, probably for kinetic reasons. By measuring the light-generated electric signal of an oriented sample, the electrogenicity of each intermediate could be determined. The electrogenicities of the first three intermediates (K, M(1), and M(2)) have small negative value, but the last three components, corresponding to the N and PR' intermediates and PR, are positive and two-orders-of-magnitude larger. These states give the major contributions to the proton translocation across the membrane. The energetic scheme of the photocycle was calculated from the temperature-dependence of the absorption kinetic signals. PMID- 12547800 TI - Time-resolved FTIR studies of sensory rhodopsin II (NpSRII) from Natronobacterium pharaonis: implications for proton transport and receptor activation. AB - The photocycle of the photophobic receptor from Natronobacterium pharaonis, NpSRII, is studied by static and time-resolved step-scan Fourier transform infrared spectroscopy. Both low-temperature static and time-resolved spectra resolve a K-like intermediate, and the corresponding spectra show little difference within the noise of the time-resolved data. As compared to intermediate K of bacteriorhodopsin, relatively large amide I bands indicate correspondingly larger distortions of the protein backbone. The time-resolved spectra identify an intermediate L-like state with surprisingly small additional molecular alterations. With the formation of intermediate M, the Schiff-base proton is transferred to the counterion Asp-75. This state is characterized by larger amide bands indicating larger distortions of the protein. We can identify a second M state that differs only in small-protein bands. Reisomerization of the chromophore to all-trans occurs with the formation of intermediate O. The accelerated decay of intermediate M caused by azide results in another red shifted intermediate with a protonated Schiff base. The chromophore in this state, however, still has 13-cis geometry. Nevertheless, the reisomerization is still as slow as under the conditions without azide. The results are discussed with respect to mechanisms of the observed proton pumping and the possible roles of the intermediates in receptor activation. PMID- 12547801 TI - Systematic improvement of protein crystals by determining the supersolubility curves of phase diagrams. AB - A systematic approach for improving protein crystals by growing them in the metastable zone using the vapor diffusion technique is described. This is a simple technique for optimization of crystallization conditions. Screening around known conditions is performed to establish a working phase diagram for the crystallization of the protein. Dilutions of the crystallization drops across the supersolubility curve into the metastable zone are then carried out as follows: the coverslips holding the hanging drops are transferred, after being incubated for some time at conditions normally giving many small crystals, over reservoirs at concentrations which normally yield clear drops. Fewer, much larger crystals are obtained when the incubation times are optimized, compared with conventional crystallization at similar conditions. This systematic approach has led to the structure determination of the light-harvesting protein C-phycocyanin to the highest-ever resolution of 1.45 A. PMID- 12547802 TI - An empirical correlation between secondary structure content and averaged chemical shifts in proteins. AB - It is shown that the averaged chemical shift (ACS) of a particular nucleus in the protein backbone empirically correlates well to its secondary structure content (SSC). Chemical shift values of more than 200 proteins obtained from the Biological Magnetic Resonance Bank are used to calculate ACS values, and the SSC is estimated from the corresponding three-dimensional coordinates obtained from the Protein Data Bank. ACS values of (1)H(alpha) show the highest correlation to helical and sheet structure content (correlation coefficient of 0.80 and 0.75, respectively); (1)H(N) exhibits less reliability (0.65 for both sheet and helix), whereas such correlations are poor for the heteronuclei. SSC estimated using this correlation shows a good agreement with the conventional chemical shift index based approach for a set of proteins that only have chemical shift information but no NMR or x-ray determined three-dimensional structure. These results suggest that even chemical shifts averaged over the entire protein retain significant information about the secondary structure. Thus, the correlation between ACS and SSC can be used to estimate secondary structure content and to monitor large scale secondary structural changes in protein, as in folding studies. PMID- 12547803 TI - The refined crystal structure of an eel pout type III antifreeze protein RD1 at 0.62-A resolution reveals structural microheterogeneity of protein and solvation. AB - RD1 is a 7-kDa globular protein from the Antarctic eel pout Lycodichthys dearborni. It belongs to type III of the four types of antifreeze proteins (AFPs) found in marine fishes living at subzero temperatures. For type III AFP, a potential ice-binding flat surface has been identified and is imbedded with side chains capable of making hydrogen bonds with a specific lattice plane on ice. So far, all crystallographic studies on type III AFPs were carried out using the Atlantic ocean pout Macrozoarces americanus as the source organism. Here we present the crystal structure of a type III AFP from a different zoarcid fish, and at an ultra-high resolution of 0.62 A. The protein fold of RD1 comprises a compact globular domain with two internal tandem motifs arranged about a pseudo dyad symmetry. Each motif of the "pretzel fold" includes four short beta-strands and a 3(10) helix. There is a novel internal cavity of 45 A(3) surrounded by eight conserved nonpolar residues. The model contains several residues with alternate conformations, and a number of split water molecules, probably caused by alternate interactions with the protein molecule. After extensive refinement that includes hydrogen atoms, significant residual electron densities associated with the electrons of peptides and many other bonds could be visualized. PMID- 12547804 TI - Effects of water soluble phosphotidylserine on bovine factor Xa: functional and structural changes plus dimerization. AB - Previous work has shown that two molecules of a soluble form of phosphatidylserine, C6PS, bind to human and bovine factor X(a). Activity measurements along with the fluorescence of active-site-labeled human factor X(a) showed that two linked sites specifically regulate the active site conformation and proteolytic activity of the human enzyme. These results imply, but cannot demonstrate, a C6PS-induced factor X(a) conformational change. The purpose of this paper is to extend these observations to bovine factor X(a) and to demonstrate that they do reflect conformational changes. We report that the fluorescence of active-site-labeled bovine factor X(a) also varied with C6PS concentration in a sigmoidal manner, whereas amidolytic activity of unlabeled enzyme varied in a simple hyperbolic fashion, also as seen for human factor X(a). C6PS induced a 70-fold increase in bovine factor X(a)'s autolytic activity, consistent with the 60-fold increase in proteolytic activity reported for human factor X(a). In addition, circular dichroism spectroscopy clearly demonstrated that C6PS binding to bovine factor X(a) induces secondary structural changes. In addition, C6PS binding to the tighter of the two sites triggered structural changes that lead to Ca(2+)-dependent dimer formation, as demonstrated by changes in intrinsic fluorescence and quantitative native gel electrophoresis. Dimerization produced further change in secondary structure, either inter- or intramolecularly. These results, along with results presented previously, support a model in which C6PS binds in a roughly sequential fashion to two linked sites whose occupancy in both human and bovine factor X(a) elicits different structural and functional responses. PMID- 12547805 TI - Force measurements of the alpha5beta1 integrin-fibronectin interaction. AB - The interaction of the alpha(5)beta(1) integrin and its ligand, fibronectin (FN), plays a crucial role in the adhesion of cells to the extracellular matrix. An important intrinsic property of the alpha(5)beta(1)/FN interaction is the dynamic response of the complex to a pulling force. We have carried out atomic force microscopy measurements of the interaction between alpha(5)beta(1) and a fibronectin fragment derived from the seventh through tenth type III repeats of FN (i.e., FN7-10) containing both the arg-gly-asp (RGD) sequence and the synergy site. Direct force measurements obtained from an experimental system consisting of an alpha(5)beta(1) expressing K562 cell attached to the atomic force microscopy cantilever and FN7-10 adsorbed on a substrate were used to determine the dynamic response of the alpha(5)beta(1)/FN7-10 complex to a pulling force. The experiments were carried out over a three-orders-of-magnitude change in loading rate and under conditions that allowed for detection of individual alpha(5)beta(1)/FN7-10 interactions. The dynamic rupture force of the alpha(5)beta(1)/FN7-10 complex revealed two regimes of loading: a fast loading regime (>10,000 pN/s) and a slow loading regime (<10,000 pN/s) that characterize the inner and outer activation barriers of the complex, respectively. Activation by TS2/16 antibody increased both the frequency of adhesion and elevated the rupture force of the alpha(5)beta(1)/wild type FN7-10 complex to higher values in the slow loading regime. In experiments carried out with a FN7-10 RGD deleted mutant, the force measurements revealed that both inner and outer activation barriers were suppressed by the mutation. Mutations to the synergy site of FN, however, suppressed only the outer barrier activation of the complex. For both the RGD and synergy deletions, the frequency of adhesion was less than that of the wild type FN7-10, but was increased by integrin activation. The rupture force of these mutants was only slightly less than that of the wild type, and was not increased by activation. These results suggest that integrin activation involved a cooperative interaction with both the RGD and synergy sites. PMID- 12547806 TI - Role of side-chain conformational entropy in transmembrane helix dimerization of glycophorin A. AB - Dimerization of the transmembrane domain of glycophorin A is mediated by a seven residue motif LIxxGVxxGVxxT through a combination of van der Waals and hydrogen bonding interactions. One of the unusual features of the motif is the large number of beta-branched amino acids that may limit the entropic cost of dimerization by restricting side-chain motion in the monomeric transmembrane helix. Deuterium NMR spectroscopy is used to characterize the dynamics of fully deuterated Val80 and Val84, two essential amino acids of the dimerization motif. Deuterium spectra of the glycophorin A transmembrane dimer were obtained using synthetic peptides corresponding to the transmembrane sequence containing either perdeuterated Val80 or Val84. These data were compared with spectra of monomeric glycophorin A peptides deuterated at Val84. In all cases, the deuterium line shapes are characterized by fast methyl group rotation with virtually no motion about the C(alpha)-C(beta) bond. This is consistent with restriction of the side chain in both the monomer and dimer due to intrahelical packing interactions involving the beta-methyl groups, and indicates that there is no energy cost associated with dimerization due to loss of conformational entropy. In contrast, deuterium NMR spectra of Met81 and Val82, in the lipid interface, reflected greater motional averaging and fast exchange between different side-chain conformers. PMID- 12547807 TI - Assembly of collagen matrices as a phase transition revealed by structural and rheologic studies. AB - We have studied the structural and viscoelastic properties of assembling networks of the extracellular matrix protein type-I collagen by means of phase contrast microscopy and rotating disk rheometry. The initial stage of the assembly is a nucleation process of collagen monomers associating to randomly distributed branched clusters with extensions of several microns. Eventually a sol-gel transition takes place, which is due to the interconnection of these clusters. We analyzed this transition in terms of percolation theory. The viscoelastic parameters (storage modulus G' and loss modulus G") were measured as a function of time for five different frequencies ranging from omega = 0.2 rad/s to 6.9 rad/s. We found that at the gel point both G' and G" obey a scaling law, with the critical exponent Delta = 0.7 and a critical loss angle being independent of frequency as predicted by percolation theory. Gelation of collagen thus represents a second order phase transition. PMID- 12547808 TI - Quantifying double-strand breaks and clustered damages in DNA by single-molecule laser fluorescence sizing. AB - Fluorescence from a single DNA molecule passing through a laser beam is proportional to the size (contour length) of the molecule, and molecules of different sizes can be counted with equal efficiencies. Single-molecule fluorescence can thus determine the average length of the molecules in a sample and hence the frequency of double-strand breaks induced by various treatments. Ionizing radiation-induced frank double-strand breaks can thus be quantified by single-molecule sizing. Moreover, multiple classes of clustered damages involving damaged bases and abasic sites, alone or in combination with frank single-strand breaks, can be quantified by converting them to double-strand breaks by chemical or enzymatic treatments. For a given size range of DNA molecules, single-molecule sizing is as or more sensitive than gel electrophoresis, and requires several orders-of-magnitude less DNA to determine damage levels. PMID- 12547809 TI - Oxygen consumption rates and oxygen concentration in molt-4 cells and their mtDNA depleted (rho0) mutants. AB - Respiratory deficient cell lines are being increasingly used to elucidate the role of mitochondria and to understand the pathophysiology of mitochondrial genetic disease. We have investigated the oxygen consumption rates and oxygen concentration in wild-type (WT) and mitochondrial DNA (mtDNA) depleted (rho(0)) Molt-4 cells. Wild-type Molt-4 cells have moderate oxygen consumption rates, which were significantly reduced in the rho(0) cells. PCMB (p chloromercurobenzoate) inhibited the oxygen consumption rates in both WT and rho(0) cells, whereas potassium cyanide decreased the oxygen consumption rates only in WT Molt-4 cells. Menadione sodium bisulfite (MSB) increased the oxygen consumption rates in both cell lines, whereas CCCP (carbonyl cyanide m chlorophenylhydrazone) stimulated the oxygen consumption rates only in WT Molt-4 cells. Superoxide radical adducts were observed in both WT and rho(0) cells when stimulated with MSB. The formation of this adduct was inhibited by PCMB but not by potassium cyanide. These results suggest that the reactive oxygen species (ROS) induced by MSB were at least in part produced via a mitochondrial independent pathway. An oxygen gradient between the extra- and intracellular compartments was observed in WT Molt-4 cells, which further increased when cells were stimulated by CCCP and MSB. The results are consistent with our earlier findings suggesting that such oxygen gradients may be a general phenomenon found in most or all cell systems under appropriate conditions. PMID- 12547810 TI - Carcinoma and SV40-transfected normal ovarian surface epithelial cell comparison by nonphotochemical hole burning. AB - Results are presented of nonphotochemical-hole-burning experiments on the mitochondrial specific dye rhodamine 800 incubated with two human ovarian surface epithelial cell lines: OSE(tsT)-14 normal cells and OV167 carcinoma cells. This dye is selective for the plasma and inner membranes of the mitochondria, as shown by confocal microscopy images. Dispersive hole-growth kinetics of zero-phonon holes are analyzed with theoretical fits, indicating that subcellular structural heterogeneity of the carcinoma cell line is lower relative to the analogous normal cell line. Broadening of holes in the presence of an applied electric field (Stark effect) was used to determine the permanent dipole moment change for the S(0)-->S(1) transition in the two cell lines. For the carcinoma cell line, the permanent dipole moment change value is a factor of 1.5 higher than for the normal cell line. It is speculated that this difference may be related to differences in mitochondrial membrane potentials in the two cell lines. PMID- 12547811 TI - Laser-induced heating in optical traps. AB - In an optical tweezers experiment intense laser light is tightly focused to intensities of MW/cm(2) in order to apply forces to submicron particles or to measure mechanical properties of macromolecules. It is important to quantify potentially harmful or misleading heating effects due to the high light intensities in biophysical experiments. We present a model that incorporates the geometry of the experiment in a physically correct manner, including heat generation by light absorption in the neighborhood of the focus, balanced by outward heat flow, and heat sinking by the glass surfaces of the sample chamber. This is in contrast to the earlier simple models assuming heat generation in the trapped particle only. We find that in the most common experimental circumstances, using micron-sized polystyrene or silica beads, absorption of the laser light in the solvent around the trapped particle, not in the particle itself, is the most important contribution to heating. To validate our model we measured the spectrum of the Brownian motion of trapped beads in water and in glycerol as a function of the trapping laser intensity. Heating both increases the thermal motion of the bead and decreases the viscosity of the medium. We measured that the temperature in the focus increased by 34.2 +/- 0.1 K/W with 1064-nm laser light for 2200-nm-diameter polystyrene beads in glycerol, 43.8 +/- 2.2 K/W for 840-nm polystyrene beads in glycerol, 41.1 +/- 0.7 K/W for 502-nm polystyrene beads in glycerol, and 7.7 +/- 1.2 K/W for 500-nm silica beads and 8.1 +/- 2.1 K/W for 444-nm silica beads in water. Furthermore, we observed that in glycerol the heating effect increased when the bead was trapped further away from the cover glass/glycerol interface as predicted by the model. We show that even though the heating effect in water is rather small it can have non negligible effects on trap calibration in typical biophysical experimental circumstances and should be taken into consideration when laser powers of more than 100 mW are used. PMID- 12547812 TI - Real-time imaging of nuclear permeation by EGFP in single intact cells. AB - The NPC is the portal for the exchange of proteins, mRNA, and ions between nucleus and cytoplasm. Many small molecules (<10 kDa) permeate the nucleus by simple diffusion through the pore, but molecules larger than 70 kDa require ATP and a nuclear localization sequence for their transport. In isolated Xenopus oocyte nuclei, diffusion of intermediate-sized molecules appears to be regulated by the NPC, dependent upon [Ca(2+)] in the nuclear envelope. We have applied real time imaging and fluorescence recovery after photobleaching to examine the nuclear pore permeability of 27-kDa EGFP in single intact cells. We found that EGFP diffused bidirectionally via the NPC across the nuclear envelope. Although diffusion is slowed approximately 100-fold at the nuclear envelope boundary compared to diffusion within the nucleus or cytoplasm, this delay is expected for the reduced cross-sectional area of the NPCs. We found no evidence for significant nuclear pore gating or block of EGFP diffusion by depletion of perinuclear Ca(2+) stores, as assayed by a nuclear cisterna-targeted Ca(2+) indicator. We also found that EGFP exchange was not altered significantly during the cell cycle. PMID- 12547813 TI - A prestressed cable network model of the adherent cell cytoskeleton. AB - A prestressed cable network is used to model the deformability of the adherent cell actin cytoskeleton. The overall and microstructural model geometries and cable mechanical properties were assigned values based on observations from living cells and mechanical measurements on isolated actin filaments, respectively. The models were deformed to mimic cell poking (CP), magnetic twisting cytometry (MTC) and magnetic bead microrheometry (MBM) measurements on living adherent cells. The models qualitatively and quantitatively captured the fibroblast cell response to the deformation imposed by CP while exhibiting only some qualitative features of the cell response to MTC and MBM. The model for CP revealed that the tensed peripheral actin filaments provide the key resistance to indentation. The actin filament tension that provides mechanical integrity to the network was estimated at approximately 158 pN, and the nonlinear mechanical response during CP originates from filament kinematics. The MTC and MBM simulations revealed that the model is incomplete, however, these simulations show cable tension as a key determinant of the model response. PMID- 12547814 TI - Structure and composition of the fusion pore. AB - Earlier studies using atomic force microscopy (AFM) demonstrated the presence of fusion pores at the cell plasma membrane in a number of live secretory cells, revealing their morphology and dynamics at nm resolution and in real time. Fusion pores were stable structures at the cell plasma membrane where secretory vesicles dock and fuse to release vesicular contents. In the present study, transmission electron microscopy confirms the presence of fusion pores and reveals their detailed structure and association with membrane-bound secretory vesicles in pancreatic acinar cells. Immunochemical studies demonstrated that t-SNAREs, NSF, actin, vimentin, alpha-fodrin and the calcium channels alpha1c and beta3 are associated with the fusion complex. The localization and possible arrangement of SNAREs at the fusion pore are further demonstrated from combined AFM, immunoAFM, and electrophysiological measurements. These studies reveal the fusion pore or porosome to be a cup-shaped lipoprotein structure, the base of which has t-SNAREs and allows for docking and release of secretory products from membrane-bound vesicles. PMID- 12547815 TI - Mechanical forces impeding exocytotic surfactant release revealed by optical tweezers. AB - The release of surfactant from alveolar type II cells is essential to lower the surface tension in the lung and to facilitate inspiration. However, the factors controlling dispersal and diffusion of this hydrophobic material are still poorly understood. Here we report that release of surfactant from the fused vesicle, termed lamellar body (LB), resisted mechanical forces applied by optical tweezers: At constant trapping force, the probability to expand LB contents, i.e., to "pull" surfactant into the extracellular fluid, increased with time after LB fusion with the plasma membrane, consistent with slow fusion pore expansion in these cells. Elevations of the cytoplasmic Ca(2+) concentration ([Ca(2+)](c)) had a similar effect. Inasmuch as surfactant did not disintegrate in the extracellular space, this method permitted for the first time the determination of elastic and recoil properties of the macromolecular complex, yielding a spring constant of approximately 12.5 pN/ micro m. This is the first functional evidence that release of hydrophobic material is mechanically impeded and occurs in an "all-or-none" fashion. This mode of release is most probably the result of cohesive forces of surfactant, combined with adhesive forces and/or retaining forces exerted by a constrictive fusion pore acting as a regulated mechanical barrier, withstanding forces up to 160 pN. In independent experiments equiaxial strain was exerted on cells without optical tweezers. Strain facilitated surfactant release from preexisting fused vesicles, consistent with the view of mechanical impediments during the release process, which can be overcome by cell strain. PMID- 12547816 TI - Molecular-level thermodynamic switch controls chemical equilibrium in sequence specific hydrophobic interaction of 35 dipeptide pairs. AB - Applying the Planck-Benzinger methodology, the sequence-specific hydrophobic interactions of 35 dipeptide pairs were examined over a temperature range of 273 333 K, based on data reported by Nemethy and Scheraga in 1962. The hydrophobic interaction in these sequence-specific dipeptide pairs is highly similar in its thermodynamic behavior to that of other biological systems. The results imply that the negative Gibbs free energy change minimum at a well-defined stable temperature, , where the bound unavailable energy, TdeltaS(o) = 0, has its origin in the sequence-specific hydrophobic interactions, are highly dependent on details of molecular structure. Each case confirms the existence of a thermodynamic molecular switch wherein a change of sign in deltaCp(o)(T)(reaction) (change in specific heat capacity of reaction at constant pressure) leads to true negative minimum in the Gibbs free energy change of reaction, deltaG(o)(T)(reaction), and hence a maximum in the related equilibrium constant, K(eq). Indeed, all interacting biological systems examined to date by Chun using the Planck-Benzinger methodology have shown such a thermodynamic switch at the molecular level, suggesting its existence may be universal. PMID- 12547817 TI - Stepwise transition of the tetra-manganese complex of photosystem II to a binuclear Mn2(micro -O)2 complex in response to a temperature jump: a time resolved structural investigation employing x-ray absorption spectroscopy. AB - In oxygenic photosynthesis, water is oxidized at a protein-cofactor complex comprising four Mn atoms and, presumably, one calcium. Using multilayers of Photosystem II membrane particles, we investigated the time course of the disassembly of the Mn complex initiated by a temperature jump from 25 degrees C to 47 degrees C and terminated by rapid cooling after distinct heating periods. We monitored polarographically the oxygen-evolution activity, the amount of the Y(D)(ox) radical and of released Mn(2+) by EPR spectroscopy, and the structure of the Mn complex by x-ray absorption spectroscopy (XAS, EXAFS). Using a novel approach to analyze time-resolved EXAFS data, we identify three distinct phases of the disassembly process: (1) Loss of the oxygen-evolution activity and reduction of Y(D)(ox) occur simultaneously (k(1) = 1.0 min(-1)). EXAFS spectra reveal the concomitant loss of an absorber-backscatterer interaction between heavy atoms separated by approximately 3.3 A, possibly related to Ca release. (2) Subsequently, two Mn(III) or Mn(IV) ions seemingly separated by approximately 2.7 A in the native complex are reduced to Mn(II) and released (k(2) = 0.18 min(-1)). The x-ray absorption spectroscopy data is highly suggestive that the two unreleased Mn ions form a di- micro -oxo bridged Mn(III)(2) complex. (3) Finally, the tightly-bound Mn(2)( micro -O)(2) unit is slowly reduced and released (k(3) = 0.014 min(-1)). PMID- 12547818 TI - State and spectral properties of chloride oscillations in pollen. AB - Pollen tube growth is a dynamic system expressing a number of oscillating circuits. Our recent work identified a new circuit, oscillatory efflux of Cl(-) anion from the pollen tube apex. Cl(-) efflux is the first ion signal found to be coupled in phase with growth oscillations. Functional analyses indicate an active role for Cl(-) flux in pollen tube growth. In this report the dynamical properties of Cl(-) efflux are examined. Phase space analysis demonstrates that the system trajectory converges on a limit cycle. Fourier analysis reveals that two harmonic frequencies characterize normal growth. Cl(-) efflux is inhibited by the channel blocker DIDS, is stimulated by hypoosmotic treatment, and is antagonized by the signal encoded in inositol 3,4,5,6-tetrakisphosphate. These perturbations induce transitions of the limit cycle to new metastable states or cause system collapse to a static attractor centered near the origin. These perturbations also transform the spectral profile, inducing subharmonic frequencies, transitions to period doubling and tripling, superharmonic resonance, and chaos. These results indicate that Cl(-) signals in pollen tubes display features that are characteristic of active oscillators that carry frequency-encoded information. A reaction network of the Cl(-) oscillator coupled to two nonlinear feedback circuits that may drive pollen tube growth oscillations is considered. PMID- 12547819 TI - Single channel analysis of the regulation of GIRK1/GIRK4 channels by protein phosphorylation. AB - G-Protein activated, inwardly rectifying potassium channels (GIRKs) are important effectors of G-protein beta/gamma-subunits, playing essential roles in the humoral regulation of cardiac activity and also in higher brain functions. G protein activation of channels of the GIRK1/GIRK4 heterooligomeric composition is controlled via phosphorylation by cyclic AMP dependent protein kinase (PKA) and dephosphorylation by protein phosphatase 2A (PP(2)A). To study the molecular mechanism of this unprecedented example of G-protein effector regulation, single channel recordings were performed on isolated patches of plasma membranes of Xenopus laevis oocytes. Our study shows that: (i) The open probability (P(o)) of GIRK1/GIRK4 channels, stimulated by coexpressed m(2)-receptors, was significantly increased upon addition of the catalytic subunit of PKA to the cytosolic face of an isolated membrane patch. (ii) At moderate concentrations of recombinant G(beta1/gamma2), used to activate the channel, P(o) was significantly reduced in patches treated with PP(2)A, when compared to patches with PKA-cs. (iii) Several single channel gating parameters, including modal gating behavior, were significantly different between phosphorylated and dephosphorylated channels, indicating different gating behavior between the two forms of the protein. Most of these changes were, however, not responsible for the marked difference in P(o) at moderate G-protein concentrations. (iv) An increase of the frequency of openings (f(o)) and a reduction of dwell time duration of the channel in the long lasting C(5) state was responsible for facilitation of GIRK1/GIRK4 channels by protein phosphorylation. Dephosphorylation by PP(2)A led to an increase of G(beta1/gamma2) concentration required for full activation of the channel and hence to a reduction of the apparent affinity of GIRK1/GIRK4 for G(beta1/gamma2). (v) Although possibly not directly the target of protein phosphorylation/dephosphorylation, the last 20 C-terminal amino acids of the GIRK1 subunit are required for the reduction of apparent affinity for the G protein by PP(2)A, indicating that they constitute an essential part of the off switch. PMID- 12547820 TI - Amplifying signal transduction specificity without multiple phosphorylation. PMID- 12547821 TI - Molecular identification of human glutamine- and ammonia-dependent NAD synthetases. Carbon-nitrogen hydrolase domain confers glutamine dependency. AB - NAD synthetase catalyzes the final step in the biosynthesis of NAD. In the present study, we obtained cDNAs for two types of human NAD synthetase (referred as NADsyn1 and NADsyn2). Structural analysis revealed in both NADsyn1 and NADsyn2 a domain required for NAD synthesis from ammonia and in only NADsyn1 an additional carbon-nitrogen hydrolase domain shared with enzymes of the nitrilase family that cleave nitriles as well as amides to produce the corresponding acids and ammonia. Consistent with the domain structures, biochemical assays indicated (i) that both NADsyn1 and NADsyn2 have NAD synthetase activity, (ii) that NADsyn1 uses glutamine as well as ammonia as an amide donor, whereas NADsyn2 catalyzes only ammonia-dependent NAD synthesis, and (iii) that mutant NADsyn1 in which Cys 175 corresponding to the catalytic cysteine residue in nitrilases was replaced with Ser does not use glutamine. Kinetic studies suggested that glutamine and ammonia serve as physiological amide donors for NADsyn1 and NADsyn2, respectively. Both synthetases exerted catalytic activity in a multimeric form. In the mouse, NADsyn1 was seen to be abundantly expressed in the small intestine, liver, kidney, and testis but very weakly in the skeletal muscle and heart. In contrast, expression of NADsyn2 was observed in all tissues tested. Therefore, we conclude that humans have two types of NAD synthetase exhibiting different amide donor specificity and tissue distributions. The ammonia-dependent synthetase has not been found in eucaryotes until this study. Our results also indicate that the carbon-nitrogen hydrolase domain is the functional domain of NAD synthetase to make use of glutamine as an amide donor in NAD synthesis. Thus, glutamine dependent NAD synthetase may be classified as a possible glutamine amidase in the nitrilase family. Our molecular identification of NAD synthetases may prove useful to learn more of mechanisms regulating cellular NAD metabolism. PMID- 12547822 TI - A Rac/Cdc42-specific exchange factor, GEFT, induces cell proliferation, transformation, and migration. AB - The Rho family of small GTPases, including Rho, Rac, and Cdc42, play essential roles in diverse cellular functions. The ability of Rho family GTPases to participate in signaling events is determined by the ratio of inactive (GDP bound) and active (GTP-bound) forms in the cell. The activation of Rho family proteins requires the exchange of bound GDP for GTP, a process catalyzed by the Dbl family of guanine nucleotide exchange factors (GEFs). The GEFs have high affinity for the guanine nucleotide-free state of the GTPases and are thought to promote GDP release by stabilizing an intermediate transition state. In this study, we have identified and characterized a new Rac/Cdc42-specific Dbl family guanine nucleotide exchange factor, named GEFT. GEFT is highly expressed in the excitable tissues, including brain, heart, and muscle. Low or very little expression was detected in other nonexcitable tissues. GEFT has specific exchange activity for Rac and Cdc42 in our in vitro GTPase exchange assays and glutathione S-transferase-PAK pull-down assays with GTP-bound Rac1 and Cdc42. Overexpression of GEFT leads to changes in cell morphology and actin cytoskeleton re organization, including the formation of membrane microspikes, filopodia, and lamilliopodia. Furthermore, expression of GEFT in NIH3T3 cells promotes foci formation, cell proliferation, and cell migration, possibly through the activation of transcriptional factors involved in cell growth and proliferation. Together, our data suggest that GEFT is a Rac/Cdc42-specific GEF protein that regulates cell morphology, cell proliferation, and transformation. PMID- 12547823 TI - Biosynthesis of 5-oxo-6,8,11,14-eicosatetraenoic acid from 5 hydroperoxyeicosatetraenoic acid in the murine macrophage. AB - 5-Oxo-6,8,11,14-eicosatetraenoic acid (5-oxo-ETE) is a metabolite of arachidonic acid shown to possess important biological activities within different cell types. In the neutrophil, a specific NADP(+)-dependent dehydrogenase utilizes 5 lipoxygenase-derived 5-hydroxy-6,8,11,14-eicosatetraenoic acid (5(S)-HETE) as the required substrate. In the present study, 5-hydroperoxy-6,8,11,14 eicosatetraenoic acid (5-HpETE), rather than 5-HETE, was found to be the biosynthetic precursor of 5-oxo-ETE in the murine macrophage. The macrophage was not able to convert 5-HETE into 5-oxo-ETE even when preincubated with phorbol ester or with other lipid hydroperoxides. The factor responsible for the conversion of 5-HpETE into 5-oxo-ETE was found predominantly in the cytosolic fraction of the macrophage, with an approximate molecular weight of 50,000 60,000, as assessed by size exclusion chromatography. Formation of 5-oxo-ETE was rapid and the catalytic protein was found to have an apparent K(m) of 5.3 microM for the eicosanoid. Furthermore, the protein could efficiently utilize 5(R,S) HpETE as substrate and was heat and protease labile. This novel pathway of 5-oxo ETE biosynthesis in the murine macrophage was consistent with reduction of a 5 hydroperoxy group to an intermediate alkoxy radical that could be subsequently oxidized to the 5-oxo product. Such a mechanism would enable racemic 5-HpETE, derived from free radical oxidation of arachidonic acid, to be efficiently converted into this potent chemotactic eicosanoid. PMID- 12547824 TI - Ca2+-dependent localization of integrin-linked kinase to cell junctions in differentiating keratinocytes. AB - Integrin complexes are necessary for proper proliferation and differentiation of epidermal keratinocytes. Differentiation of these cells is accompanied by down regulation of integrins and focal adhesions as well as formation of intercellular adherens junctions through E-cadherin homodimerization. A central component of integrin adhesion complexes is integrin-linked kinase (ILK), which can induce loss of E-cadherin expression and epithelial-mesenchymal transformation when ectopically expressed in intestinal and mammary epithelia. In cultured primary mouse keratinocytes, we find that ILK protein levels are independent of integrin expression and signaling, since they remain constant during Ca(2+)-induced differentiation. In contrast, keratinocyte differentiation is accompanied by marked reduction in kinase activity in ILK immunoprecipitates and altered ILK subcellular distribution. Specifically, ILK distributes in close apposition to actin fibers along intercellular junctions in differentiated but not in undifferentiated keratinocytes. ILK localization to cell-cell borders occurs independently of integrin signaling and requires Ca(2+) as well as an intact actin cytoskeleton. Further, and in contrast to what is observed in other epithelial cells, ILK overexpression in differentiated keratinocytes does not promote E-cadherin down-regulation and epithelial-mesenchymal transition. Thus, novel tissue-specific mechanisms control the formation of ILK complexes associated with cell-cell junctions in differentiating murine epidermal keratinocytes. PMID- 12547825 TI - 17-epiestriol, an estrogen metabolite, is more potent than estradiol in inhibiting vascular cell adhesion molecule 1 (VCAM-1) mRNA expression. AB - 17-beta estradiol (17-beta E(2)) attenuates the expression of vascular cell adhesion molecule 1 (VCAM-1) in vivo at physiological levels (pg/ml), whereas supraphysiological concentrations of 17-beta E(2) (ng/ml) are required in vitro. We assessed whether a metabolite of estrogen, which could only be generated in vivo, might be a more potent inhibitor of VCAM-1 expression and thereby explain this discrepancy. We report here that 17-epiestriol, an estrogen metabolite and a selective estrogen receptor (ER) beta agonist, is approximately 400x more potent than 17-beta E(2) in suppressing tumor necrosis factor (TNF) alpha-induced VCAM-1 mRNA as well as protein expression in human umbilical vein endothelial cells. Genistein, an ERbeta agonist, at low concentrations (1 and 10 nm) also suppressed TNFalpha-induced VCAM-1 mRNA expression. These actions of 17-epiestriol and genistein were significantly attenuated in the presence of the estrogen receptor antagonist ICI-182780. Other estrogenic compounds such as ethinyl estradiol and estrone did not have any effect on TNFalpha-induced VCAM-1 expression at the concentrations tested. We further show that, 1) 17-epiestriol induces the expression of endothelial nitric-oxide synthase mRNA and protein, 2) 17 epiestriol prevents TNFalpha-induced migration of NFkappaB into the nucleus, 3) N(G)-nitro-l-arginine methyl ester, an inhibitor of NO synthesis, abolishes 17 epiestriol-mediated inhibition of TNFalpha-induced VCAM-1 expression and migration of NFkappaB from the cytoplasm to the nucleus. Our results indicate that 17-epiestriol is more potent than 17-beta E(2) in suppressing TNFalpha induced VCAM-1 expression and that this action is modulated at least in part through NO. PMID- 12547826 TI - Tumor promoter arsenite stimulates histone H3 phosphoacetylation of proto oncogenes c-fos and c-jun chromatin in human diploid fibroblasts. AB - Although epidemiological studies have long established that inorganic arsenic is a potent human carcinogen, the underlying mechanisms are still poorly understood. Recent studies suggest that inorganic arsenic may act as a tumor promoter by perturbing key signaling transduction pathways. We have shown previously that arsenite can potently activate the mitogen-activated protein kinase cascades and induce the expression of proliferation-associated genes, including proto oncogenes c-jun and c-fos. In order to elucidate further the molecular mechanisms underlying its tumor-promoting properties, we investigated the signaling events involved in arsenite-mediated induction of c-fos and c-jun. We found that induction of both c-fos and c-jun by arsenite can be substantially inhibited by the MEK- selective inhibitor U0126, suggesting that the ERK pathway is critically involved in their up-regulation. Interestingly, arsenite dramatically induced the phosphorylation and acetylation of histone H3 preceding the induction of mRNAs encoding c-fos and c-jun. Finally, chromatin immunoprecipitation assays revealed that arsenite treatment markedly induced the phosphorylation/acetylation of histone H3 associated with the c-fos and c-jun genes through an ERK-dependent pathway. Our results strongly suggest that arsenic-triggered alterations in chromatin structure perturb specific gene transcription, including that of proto oncogenes c-jun and c-fos, and may thereby contribute to the carcinogenic process. PMID- 12547827 TI - Crystal structure of PTP1B complexed with a potent and selective bidentate inhibitor. AB - Protein-tyrosine phosphatase 1B (PTP1B) has been implicated as an important regulator in several signaling pathways including those initiated by insulin and leptin. Potent and specific PTP1B inhibitors could serve as useful tools in elucidating the physiological functions of PTP1B and may constitute valuable therapeutics in the treatment of several human diseases. We have determined the crystal structure of PTP1B in complex with compound 2, the most potent and selective PTP1B inhibitor reported to date. The structure at 2.15-A resolution reveals that compound 2 simultaneously binds to the active site and a unique proximal noncatalytic site formed by Lys-41, Arg-47, and Asp-48. The structural data are further corroborated by results from kinetic analyses of the interactions of PTP1B and its site-directed mutants with compound 2 and several of its variants. Although many of the residues important for interactions between PTP1B and compound 2 are not unique to PTP1B, the combinations of all contact residues differ between PTP isozymes, which provide a structural basis for potent and selective PTP1B inhibition. Our data further suggest that potent, yet highly selective, PTP1B inhibitory agents can be acquired by targeting the area defined by residues Lys-41, Arg-47, and Asp-48, in addition to the previously identified second aryl phosphate-binding pocket. PMID- 12547828 TI - Expression of kinase-inactive c-Src delays oxidative stress-induced disassembly and accelerates calcium-mediated reassembly of tight junctions in the Caco-2 cell monolayer. AB - The activity of Src kinases appears to play a role in both assembly and disassembly of tight junction. However, the role of a specific isoform of Src kinase in regulation of tight junction is not known. In the present study the role of c-Src in regulation of epithelial tight junction was investigated in Caco 2 cell monolayers. Oxidative stress (xanthine oxidase + xanthine) induced an activation and membrane translocation of c-Src. The oxidative stress-induced decrease in transepithelial electrical resistance, increase in inulin permeability, and redistribution of occludin and ZO-1 from the intercellular junctions were prevented by PP2. The rates of oxidative stress-induced activation of c-Src, tyrosine phosphorylation of ZO-1 and beta-catenin, decrease in resistance, increase in permeability to inulin, and redistribution of occludin and ZO-1 were significantly greater in cells transfected with wild type c-Src, whereas it was low in cells transfected with kinase-inactive c-SrcK297R mutant, when compared with those in empty vector-transfected cells. The rates of recovery of resistance, increase in barrier to inulin, and reorganization of occludin and ZO-1 into the intercellular junctions during the calcium-induced reassembly of tight junction were much greater in Caco-2 cells transfected with c-SrcK297R as compared with those in cells transfected with empty vector or wild type c-Src. These results show that the dominant-negative expression of kinase-inactive c-Src delays the oxidative stress-induced disruption of tight junction and accelerates calcium-induced assembly of tight junction in Caco-2 cells and demonstrate that oxidative stress-induced disruption of tight junction is mediated by the activation of c-Src. PMID- 12547830 TI - Stability of dark state rhodopsin is mediated by a conserved ion pair in intradiscal loop E-2. AB - The rhodopsin crystal structure reveals that intradiscal loop E-2 covers the 11 cis-retinal, creating a "retinal plug." Recently, we noticed the ends of loop E-2 are linked by an ion pair between residues Arg-177 and Asp-190, near the highly conserved disulfide bond. This ion pair appears biologically significant; it is conserved in almost all vertebrate opsins and may occur in other G-protein coupled receptors. We report here that the Arg-177/Asp-190 ion pair is critical for the folding and stability of dark state rhodopsin. We find ion pair mutants that regenerate with retinal are functionally and spectrally wild-type-like yet thermally unstable in their dark state because of rapid hydrolysis of the retinal Schiff base linkage. Surprisingly, Arrhenius analysis indicates that the activation energies for the hydrolysis process are similar between the ion pair mutants and wild-type rhodopsin. Furthermore, the ion pair mutants do not show increased reactivity toward hydroxylamine, suggesting that their instability is not caused by an increased exposure to bulk solvent. Our results indicate that the loop E-2 ion pair is important for rhodopsin stability and thus suggest that retinitis pigmentosa observed in patients with Asp-190 mutations may in part be the result of thermally unstable rhodopsin proteins. PMID- 12547829 TI - Ca2+-independent phospholipase A2 is a novel determinant of store-operated Ca2+ entry. AB - Store-operated cation (SOC) channels and capacitative Ca(2+) entry (CCE) play very important role in cellular function, but the mechanism of their activation remains one of the most intriguing and long lasting mysteries in the field of Ca(2+) signaling. Here, we present the first evidence that Ca(2+)-independent phospholipase A(2) (iPLA(2)) is a crucial molecular determinant in activation of SOC channels and store-operated Ca(2+) entry pathway. Using molecular, imaging, and electrophysiological techniques, we show that directed molecular or pharmacological impairment of the functional activity of iPLA(2) leads to irreversible inhibition of CCE mediated by nonselective SOC channels and by Ca(2+)-release-activated Ca(2+) (CRAC) channels. Transfection of vascular smooth muscle cells (SMC) with antisense, but not sense, oligonucleotides for iPLA(2) impaired thapsigargin (TG)-induced activation of iPLA(2) and TG-induced Ca(2+) and Mn(2+) influx. Identical inhibition of TG-induced Ca(2+) and Mn(2+) influx (but not Ca(2+) release) was observed in SMC, human platelets, and Jurkat T lymphocytes when functional activity of iPLA(2) was inhibited by its mechanism based suicidal substrate, bromoenol lactone (BEL). Moreover, irreversible inhibition of iPLA(2) impaired TG-induced activation of single nonselective SOC channels in SMC and BAPTA (1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid)-induced activation of whole-cell CRAC current in rat basophilic leukemia cells. Thus, functional iPLA(2) is required for activation of store-operated channels and capacitative Ca(2+) influx in wide variety of cell types. PMID- 12547831 TI - A novel conotoxin from Conus betulinus, kappa-BtX, unique in cysteine pattern and in function as a specific BK channel modulator. AB - A novel conotoxin, kappa-conotoxin (kappa-BtX), has been purified and characterized from the venom of a worm-hunting cone snail, Conus betulinus. The toxin, with four disulfide bonds, shares no sequence homology with any other conotoxins. Based on a partial amino acid sequence, its cDNA was cloned and sequenced. The deduced sequence consists of a 26-residue putative signal peptide, a 31-residue mature toxin, and a 13-residue extra peptide at the C terminus. The extra peptide is cleaved off by proteinase post-processing. All three Glu residues are gamma-carboxylated, one of the two Pro residues is hydroxylated at position 27, and its C-terminal residue is Pro-amidated. The monoisotopic mass of the toxin is 3569.0 Da. Electrophysiological experiments show that: 1) among voltage-gated channels, kappa-BtX is a specific modulator of K(+) channels; 2) among the K channels, kappa-BtX specifically up-modulates the Ca(2+)- and voltage sensitive BK channels (252 +/- 47%); 3) its EC(50) is 0.7 nm with a single binding site (Hill = 0.88); 4) the time constant of wash-out is 8.3 s; and 5) kappa-BtX has no effect on single channel conductance, but increases the open probability of BK channels. It is concluded that kappa-BtX is a novel specific biotoxin against BK channels. PMID- 12547832 TI - The lipidation by hepatocytes of human apolipoprotein A-I occurs by both ABCA1 dependent and -independent pathways. AB - The pathways of hepatic intra- and peri-cellular lipidation of apolipoprotein A-I (apoA-I) were studied by infecting primary mouse hepatocytes from either apoA-I deficient or ABCA1-deficient mice with a recombinant adenovirus expressing the human apoA-I (hapoA-I) cDNA (endo apoA-I) or incubating the hepatocytes with exogenously added hapoA-I (exo apoA-I) and examining the hapoA-I-containing lipoproteins formed. The cells, maintained in serum-free medium, were labeled with [(3)H]choline, and the cell medium was separated by fast protein liquid chromatography or immunoprecipitated to quantify labeled choline phospholipids specifically associated with hapoA-I. With the apoA-I-deficient hepatocytes, the high density lipoprotein fraction formed with endo apoA-I contained proportionally more phospholipids than that formed with exo apoA-I. However, the lipoprotein size and electrophoretic mobility and phospholipid profiles were similar for exo apoA-I and endo apoA-I. Taken together, these data demonstrate that a significant proportion of hapoA-I is secreted from hepatocytes in a phospholipidated state but that hapoA-I is also phospholipidated peri-cellularly. With primary hepatocytes from ABCA1-deficient mice, the expression and net secretion of adenoviral-generated endogenous apoA-I was unchanged compared with control mice, but (3)H-phospholipids associated with endo apoA-I and exo apoA-I decreased by 63 and 25%, respectively. The lipoprotein size and electrophoretic migration and their phospholipid profiles remained unchanged. In conclusion, we demonstrated that intracellular and peri-cellular lipidation of apoA-I represent distinct and additive pathways that may be regulated independently. Hepatocyte expression of ABCA1 is central to the lipidation of newly synthesized apoA-I but also contributes to the lipidation of exogenous apoA-I. However, a significant basal level of phospholipidation occurs in the absence of ABCA1. PMID- 12547833 TI - 22R-hydroxycholesterol and 9-cis-retinoic acid induce ATP-binding cassette transporter A1 expression and cholesterol efflux in brain cells and decrease amyloid beta secretion. AB - The ATP-binding cassette transporter A1 (ABCA1) is a major regulator of peripheral cholesterol efflux and plasma high density lipoprotein metabolism. In adult rat brain we found high expression of ABCA1 in neurons in the hypothalamus, thalamus, amygdala, cholinergic basal forebrain, and hippocampus. Large neurons of the cholinergic nucleus basalis together with CA1 and CA3 pyramidal neurons were among the most abundantly immunolabeled neurons. Glia cells were largely negative. Because cholesterol homeostasis may have an essential role in central nervous system function and neurodegeneration, we examined ABCA1 expression and function in different brain cell types using cultures of primary neurons, astrocytes, and microglia isolated from embryonic rat brain. The basal ABCA1 mRNA and protein levels detected in these cell types were increased markedly after exposure to oxysterols and 9-cis-retinoic acid, which are ligands for the nuclear hormone liver X receptors and retinoic X receptors, respectively. Functionally, the increased ABCA1 expression caused by these ligands was followed by elevated apoA-I- and apoE-specific cholesterol efflux in neurons and glia. In non-neuronal and neuronal cells overexpressing a human Swedish variant of amyloid precursor protein, 22R-hydroxycholesterol and 9-cis-retinoic acid induced ABCA1 expression and increased apoA-I-mediated cholesterol efflux consequently decreasing cellular cholesterol content. More importantly, we demonstrated that these ligands alone or in combination with apoA-I caused a substantial reduction in the stability of amyloid precursor protein C-terminal fragments and decreased amyloid beta production. These effects of 22R-hydroxycholesterol may provide a novel strategy to decrease amyloid beta secretion and consequently reduce the amyloid burden in the brain. PMID- 12547834 TI - Natriuretic peptide receptor A activation stabilizes a membrane-distal dimer interface. AB - We have shown previously (Rondeau, J.-J., McNicoll, N., Gagnon, J., Bouchard, N., Ong, H., and De Lean, A. (1995) Biochemistry 34, 2130-2136) that atrial natriuretic peptide (ANP) stabilizes a dimeric form of the natriuretic peptide receptor A (NPRA) by simultaneously interacting with both receptor subunits. However, the first crystallographic study of unliganded NPRA extracellular domain documented a V-shaped dimer involving a membrane-proximal dimer interface and separate binding sites for ANP on each monomer. We explored the possibility of an alternative A-shaped dimer involving a membrane-distal dimer interface by substituting an unpaired solvent-exposed cysteine for Trp(74) in the amino terminal lobe of full-length NPRA. The predicted spacing between Trp(74) from both subunits drastically differs, depending on whether the V-shaped (84 A) or the A-shaped (8 A) dimer model is considered. In contrast with the expected results for the reported V-shaped dimer, the NPRA(W74C) mutant was constitutively covalently dimeric. Also, the subunits spontaneously reassociated following transient disulfide reduction by dithiothreitol and reoxidation. However, ANP could neither bind to nor activate NPRA(W74C). Permanent disulfide opening by reduction with dithiothreitol and alkylation with N-ethylmaleimide rescued ANP binding to NPRA(W74C). The NPRA mutant could be maintained as a covalent dimer while preserving its function by crosslinking with the bifunctional alkylating agent phenylenedimaleimides (PDM), the ortho-substituted oPDM being more efficient than mPDM or pPDM. These results indicate that the membrane-distal lobe of the NPRAM extracellular domains are dynamically interfacing in the unliganded state and that ANP binding stabilizes the receptor dimer with more stringent spacing at the dimer interface. PMID- 12547836 TI - Molecular determinants of the stereoselectivity of agonist activity of estrogen receptors (ER) alpha and beta. AB - The two known estrogen receptors, ERalpha and ERbeta, are hormone-inducible transcription factors that have distinct roles in regulating cell proliferation and differentiation. Previously, our laboratory demonstrated that ERalpha exhibits stereoselective ligand binding and transactivation for several structural derivatives and metabolites of the synthetic estrogen diethylstilbestrol. We have previously described the properties of indenestrol A (IA) enantiomers on ERalpha. In the study presented here, the estrogenic properties of the S and R enantiomers of IA, IA-S and IA-R, respectively, were expanded to examine the activity in different cell and promoter contexts using ERalpha and ERbeta. Using human cell lines stably expressing either ERalpha or ERbeta, we found that IA-S was a more potent activator of transcription than IA-R through ERalpha in human endometrial Ishikawa and breast MDA-MB 231 (MDA) cells. Interestingly, IA-R was more potent on ERbeta when compared with ERalpha in MDA, but not in Ishikawa cells, and IA-R exhibited equally low binding affinities to ERalpha and ERbeta in vitro in contrast to its cell line-dependent preferential activation of ERbeta. Alignment of the protein structures of the ligand-binding domains of ERalpha and ERbeta revealed one mismatched residue, Leu-384 in ERalpha and Met-283 in ERbeta, which may be responsible for making contact with the methyl substituent at the chiral carbon of IA-S and IA-R. Mutagenesis and exchange of this one residue showed that the binding of IA-R and IA-S was not affected by this mutation in ERalpha and ERbeta. However, in transactivation studies, IA-R showed higher potency in activating L384M-mutated ERalpha and wild type ERbeta compared with wild-type ERalpha and M283L-mutated ERbeta in all cell and promoter contexts examined. Furthermore, IA-R-bound ERalpha L384M and wild type ERbeta displayed enhanced interactions with the nuclear receptor interaction domains of the coactivators SRC-1 and GRIP1. These data demonstrate that a single residue in the ligand-binding domain determines the stereoselectivity of ERalpha and ERbeta for indenestrol ligands and that IA-R shows cell type selectivity through ERbeta. PMID- 12547835 TI - Smads 2 and 3 are differentially activated by transforming growth factor-beta (TGF-beta ) in quiescent and activated hepatic stellate cells. Constitutive nuclear localization of Smads in activated cells is TGF-beta-independent. AB - Hepatic stellate cells are the primary cell type responsible for matrix deposition in liver fibrosis, undergoing a process of transdifferentiation into fibrogenic myofibroblasts. These cells, which undergo a similar transdifferentiation process when cultured in vitro, are a major target of the profibrogenic agent transforming growth factor-beta (TGF-beta). We have studied activation of the TGF-beta downstream signaling molecules Smads 2, 3, and 4 in hepatic stellate cells (HSC) cultured in vitro for 1, 4, and 7 days, with quiescent, intermediate, and fully transdifferentiated phenotypes, respectively. Total levels of Smad4, common to multiple TGF-beta superfamily signaling pathways, do not change as HSC transdifferentiate, and the protein is found in both nucleus and cytoplasm, independent of treatment with TGF-beta or the nuclear export inhibitor leptomycin B. TGF-beta mediates activation of Smad2 primarily in early cultured cells and that of Smad3 primarily in transdifferentiated cells. The linker protein SARA, which is required for Smad2 signaling, disappears with transdifferentiation. Additionally, day 7 cells demonstrate constitutive phosphorylation and nuclear localization of Smad 2, which is not affected by pretreatment with TGF-beta-neutralizing antibodies, a type I TGF-beta receptor kinase inhibitor, or activin-neutralizing antibodies. These results demonstrate essential differences between TGF-beta-mediated signaling pathways in quiescent and in vitro transdifferentiated hepatic stellate cells. PMID- 12547837 TI - Postnatal maturation of vagal respiratory reflexes in preterm and full-term lambs. AB - The postnatal development of ventilatory reflexes originating from bronchopulmonary receptors was assessed in preterm vs. full-term lambs. Ventilation and arterial pressure were repeatedly measured in 10 preterm (gestational age, 132 days) and 7 full-term lambs without sedation from day 1 to day 42. The Hering-Breuer inhibitory reflex (slowly adapting stretch receptors) was assessed by the increase in expiratory time during end-inspiratory occlusion. The pulmonary chemoreflex (C-fiber endings) was assessed by the initial apnea + bradycardia + systemic hypotension and the secondary tachypnea after capsaicin intravenous injection. Results show the following. 1) Premature birth did not modify the maturation of the Hering-Breuer reflex. 2) Whereas a classic pulmonary chemoreflex was observed in the very first hours of life in preterm lambs, the tachypneic component of this reflex was weaker than in full-term lambs on day 1. 3) Premature birth led to a reversed postnatal maturation of this tachypneic response (tendency to increase with postnatal age). Our findings suggest that premature birth in lambs modifies postnatal maturation of the pulmonary chemoreflex. PMID- 12547838 TI - Influence of surface chemistry and topography of particles on their immersion into the lung's surface-lining layer. AB - Inhaled and deposited spherical particles, 1-6 micrometer in diameter and of differing surface chemistry and topography, were studied in hamster intrapulmonary conducting airways and alveoli by electron microscopy. Polystyrene and Teflon particles, as well as puffball spores, were found submersed in the aqueous lining layer and adjacent to epithelial cells. The extent of particle immersion promoted by a surfactant film was assessed in a "floating-drop-surface balance" by light microscopy. Teflon and polystyrene spheres were immersed into the subphase by 50-60% at film surface tensions of 25 and 30 mJ/m(2), respectively, and totally submersed at 15 and 25 mJ/m(2), respectively. Puffball spores were immersed by approximately 50% at 22 mJ/m(2) and totally submersed at film surface tensions of 0.9). At the same relative intensity (60% Vo(2 peak)), leg norepinephrine spillover rates were approximately twofold higher in the older men (P = 0.38). Exercise-induced increases in leg arterial-venous O(2) difference were identical between groups (P > 0.9) because both arterial and venous O(2) contents were lower in the older vs. younger men. These results suggest that the ability to augment active limb blood flow and O(2) extraction during submaximal large muscle mass exercise is not impaired but is well preserved with age in healthy men who are normally active. PMID- 12547842 TI - Canine model of nasal congestion and allergic rhinitis. AB - The ragweed- and histamine-induced decreases in nasal patency in cohorts of ragweed-sensitized and nonsensitized dogs were assessed. The volume of nasal airways (V(NA)) was assessed by acoustic rhinometry and resistance to airflow (R(NA)) by anterior rhinomanometry. Histamine delivered to the nasal passages of five dogs caused a rapid and prolonged increase in R(NA) (0.75 +/- 0.26 to 3.56 +/- 0.50 cmH(2)O. l(-1). min), an effect that was reversed by intranasal delivery of aerosolized phenylephrine. Ragweed challenge in five ragweed-sensitized dogs increased R(NA) from 0.16 +/- 0.02 to 0.53 +/- 0.07 cmH(2)O. l(-1). min and decreased V(NA) from 12.5 +/- 1.9 to 3.9 +/- 0.3 cm(3), whereas administration of saline aerosol neither increased R(NA) nor decreased V(NA). Prior administration of d-pseudoephedrine (30 mg po) attenuated the ragweed-induced increase in R(NA) and decrease in V(NA). Ragweed challenge changed neither R(NA) nor V(NA) in four nonsensitized dogs. Mediator-induced nasal congestion and allergen-induced allergic rhinitis in ragweed-sensitized dogs, which exhibit symptoms similar to human disease, can be used in the evaluation of safety and efficacy of antiallergic activity of potential drugs. PMID- 12547843 TI - Regional differences in effects of exercise training on contractile and biochemical properties of rat cardiac myocytes. AB - Myocardial function is enhanced by endurance exercise training, but the cellular mechanisms underlying this improved function remain unclear. A number of studies have shown that the characteristics of cardiac myocytes vary across the width of the ventricular wall. We have previously shown that endurance exercise training alters the Ca2+ sensitivity of tension as well as contractile protein isoform expression in rat cardiac myocytes. We tested the hypothesis that these effects of training are not uniform across the ventricular wall but are more pronounced in the subendocardial (Endo) region of the myocardium. Female Sprague-Dawley rats were divided into sedentary control (C) and exercise trained (T) groups. T rats underwent 11 wk of progressive treadmill exercise. Myocytes were isolated from the Endo region of the myocardium and from the subepicardial (Epi) region of both T and C hearts. We found an increase in the Ca2+ sensitivity of tension in T cells compared with C cells, but this difference was larger in the Endo cells than in the Epi cells. In addition, we found a training-induced increase in atrial myosin light chain 1 (aMLC1) expression that was larger in the Endo compared with Epi samples. We conclude that effects of exercise training on myocyte contractile and biochemical properties are greater in myocytes from the Endo region of the myocardium than those from the Epi region. In addition, these results provide evidence that the increase in aMLC1 expression may be responsible for some of the training-induced increase in myocyte Ca2+ sensitivity of tension. PMID- 12547844 TI - Activation among the elbow flexor muscles differs when maintaining arm position during a fatiguing contraction. AB - Twenty-four men (n = 11) and women (n = 13) supported an inertial load equivalent to 20% of the maximum voluntary contraction force with the elbow flexor muscles for as long as possible while maintaining a constant elbow angle at 90 degrees. Endurance time did not differ on the three occasions that the task was performed (320 +/- 149 s; P > 0.05), and there was no difference between women (360 +/- 168 s) and men (273 +/- 108 s; P = 0.11). The rate of increase in average electromyogram (EMG) for the elbow flexor muscles was similar across sessions (P > 0.05). However, average EMG during the fatiguing task increased for the long head of biceps brachii, brachioradialis, and brachialis (P < 0.05) but not for the short head of biceps brachii. Furthermore, the average EMG for the brachialis was greater at the start and end of the contraction compared with the other elbow flexor muscles. The rate of bursts in EMG activity increased during the fatiguing contraction and was greater in brachialis (1.0 +/- 0.2 bursts/min) compared with the other elbow flexor muscles (0.5 +/- 0.1 bursts/min). The changes in the standard deviation of acceleration, mean arterial pressure, and heart rate during the fatiguing contractions were similar across sessions. These findings indicate that the EMG activity, which reflects the net excitatory and inhibitory input received by the motoneurons in the spinal cord, was not adaptable over repeat sessions for the maintain-position task. Furthermore, these results contrast those from a previous study (Hunter SK and Enoka RM. J Appl Physiol 94: 108-118, 2003) when the goal of the isometric contraction was to maintain a constant force. These results, from a series of studies on the elbow flexor muscles, indicate that the type of load supported during the fatiguing contraction influences the extent to which endurance time can change with repeat performances of the task. PMID- 12547845 TI - Concurrent exercise prevents high-fat-diet-induced macrovesicular hepatic steatosis. AB - The purpose of the present study was to assess the effect of an exercise training program conducted concurrently with a high-fat (HF)-diet regimen on the induction of hepatic steatosis. Two groups of rats were fed either a standard (SD) or a HF (40% kcal) diet for 8 wk and were additionally assigned either to a sedentary (Sed) or a treadmill-trained (TR) group. Training (5 days/wk) was initiated at the same time as the HF diet and was progressively increased, reaching 60 min at 26 m/min, 10% grade, for the last 4 wk. At the end of the 8-wk period, HF-Sed rats exhibited approximately 72% higher liver triacylglycerol concentration than SD-Sed rats (means +/- SE: 17.15 +/- 1.5 vs. 9.98 +/- 1.0 mg/g; P < 0.01). Histological quantification of lipid infiltration, with the use of an image analysis computing system, revealed that, although fat was mainly stored as microvesicles (<1 microm(2)), the HF-diet-induced hepatic steatosis occurred via the accumulation of macrovesicles (>1 microm(2)). Concurrent exercise training completely prevented the HF-diet-induced hepatic steatosis. The surface area of liver parenchyma infiltrated by lipid vacuoles was similar in HF-TR as in SD-Sed rats (26.4 +/- 1.8 vs. 29.3 +/- 5.9 x 10(3) microm(2)/200,000 microm(2) of liver parenchyma, respectively; P > 0.05). The different states of liver lipid infiltration after the HF diet in Sed and TR rats were associated with similar changes in plasma free fatty acids and glycerol, as well as with similar changes in fat pad weights, but not with plasma triacylglycerol levels. It is concluded that, after a HF-diet regimen of 8 wk in rats, hepatic steatosis occurs primarily via the accumulation of lipid as macrovesicles. Exercise training pursued at the same time completely prevents the HF-diet-induced macrovesicular hepatic steatosis. PMID- 12547846 TI - Did publication of a clinical practice guideline recommendation to screen for type 2 diabetes in women with gestational diabetes change practice? AB - OBJECTIVE: To determine whether women with previous gestational diabetes mellitus (GDM) were screened postpartum for type 2 diabetes according to the Canadian Diabetes Association (CDA) guidelines. RESEARCH DESIGN AND METHODS: The 1998 CDA guidelines recommend that all women diagnosed with GDM be screened postpartum for type 2 diabetes using a 2-h 75-g oral glucose tolerance test (OGTT). The impact of and compliance with this expert opinion-based recommendation is unknown. All women who delivered at the Ottawa Hospital in 1997 (pre-guideline) and 2000 (post guideline) with confirmed GDM were identified. Using population-based administrative databases, we determined the proportion of these women who had an OGTT, serum glucose test, or glycated hemoglobin (GHb) test in the first postpartum year. Women who had not undergone any blood work were excluded. RESULTS: There were 131 women in 1997 and 123 women in 2000 with confirmed GDM. Of these, only 69 women in 1997 and 52 women in 2000 had blood work recorded in the database. None of these women had an OGTT performed in either period. We found a significant increase in the measurement of serum glucose (50 women pre guideline [72.1%], 48 women post-guideline [92.3%], P < 0.05) and GHb (8 women pre-guideline [11.6%], 20 women post-guideline [38.5%], P < 0.01). CONCLUSIONS: In our region, physicians are not following the CDA recommendations to screen women with GDM postpartum with an OGTT. However, we did find a significant increase in the measurement of serum glucose and GHb. Publication of expert opinion-based guidelines did not change the postpartum use of an OGTT in these women but may have increased the use of less reliable screening tests for type 2 diabetes. PMID- 12547847 TI - Acarbose improves glycemic control in overweight type 2 diabetic patients insufficiently treated with metformin. AB - OBJECTIVE: To investigate the efficacy and safety of acarbose as add-on therapy in overweight type 2 patients with diabetes inadequately controlled by metformin. RESEARCH DESIGN AND METHODS: This study adopted a multicenter, randomized, double blind, placebo-controlled, parallel group design. After a 4-week placebo run-in period, subjects were randomized to either acarbose (titrated up to 100 mg b.i.d.) or placebo. The primary efficacy variable was the change in HbA(1c) from baseline to the end of the 24-week treatment period. Change in fasting blood glucose was assessed as a secondary efficacy parameter. RESULTS: The intention-to treat analysis from baseline to week 24 (81 patients for HbA(1c) and 82 for fasting blood glucose) showed statistically significant differences between acarbose and placebo treatment in HbA(1c) (1.02%; 95% CI 0.543-1.497; P = 0.0001) and fasting blood glucose (1.132 mmol/l; 95% CI 0.056-2.208; P = 0.0395) (adjusted least square means). In all, 18 patients (47%) in the acarbose group were classified as responders with a > or =5% reduction in HbA(1c) (relative to baseline) at the end point compared to 6 (14%) in the placebo group (P = 0.001). The safety profiles were similar for both treatment groups except for the higher incidence of gastrointestinal side effects during acarbose therapy. CONCLUSIONS: The addition of acarbose to metformin monotherapy provides an efficacious and safe alternative for glycemic improvement in overweight type 2 patients inadequately controlled by metformin alone. PMID- 12547848 TI - Serum ACE predicts severe hypoglycemia in children and adolescents with type 1 diabetes. AB - OBJECTIVE: To investigate whether risk of severe hypoglycemia is related to serum (S) ACE level during intensive treatment in type 1 diabetic children. RESEARCH DESIGN AND METHODS: A cohort of 86 intensively treated type 1 diabetic patients was studied during 1999-2000. In 1999, the age range was 7-19 years (median 12.8), diabetes duration was 1.2-14.7 years (5.3), insulin dose was 0.4-1.7 units x kg(-1) x 24 h(-1) (1.0), and the HbA(1c) year mean was 4.7-10.2% (6.8). HbA(1c), insulin doses, and events of severe hypoglycemia (needing assistance from another person) were prospectively registered at regular visits, scheduled quarterly. S-ACE was determined once. RESULTS: Severe hypoglycemia was correlated to S-ACE (r = 0.22, 95% CI 0.01-0.41, P = 0.0093). The square root of severe hypoglycemia was correlated to S-ACE (r = 0.27, 95% CI 0.06-0.45, P = 0.0093). Patients with S-ACE at the median level or above (n = 44) reported a mean of 3.0 yearly events of severe hypoglycemia compared with 0.5 events in patients with S ACE lower than the median (n = 42) (P = 0.0079). Of the patients with an S-ACE at the median level or above, 27 (61%) reported severe hypoglycemia, compared with 17 (40%) patients with an S-ACE lower than the median (P = 0.0527). Insulin dose, HbA(1c), age, onset age, duration, C-peptide, and sex did not differ between these two groups. S-ACE was negatively correlated with age (r = -0.27, 95% CI 0.46 to 0.07, P = 0.0265) but not with HbA(1c), duration, or blood pressure. CONCLUSIONS: The elevated rate of severe hypoglycemia among patients with higher S-ACE suggests, among other factors, that a genetic determinant for severe hypoglycemia exists. Further evaluation is needed before the clinical usefulness of this test can be elucidated. PMID- 12547849 TI - Sildenafil citrate for treatment of erectile dysfunction in men with type 1 diabetes: results of a randomized controlled trial. AB - OBJECTIVE: In the 5-10% of diabetic men with type 1 diabetes, erectile dysfunction (ED) may be a particularly common and unwanted complication. This is the first study focusing exclusively on the effects of sildenafil in men with type 1 diabetes and ED. RESEARCH DESIGN AND METHODS: A total of 188 patients were entered into a double-blind, placebo-controlled, parallel-group, flexible-dose study and were randomized to receive sildenafil (25-100 mg; n = 95) or placebo (n = 93) for 12 weeks. Efficacy was evaluated using questions three (Q3; achieving an erection) and four (Q4; maintaining an erection) from the International Index of Erectile Function (IIEF), a global efficacy question (GEQ; "Did treatment improve your erections?"), and a patient event log of sexual activity. RESULTS: Improvements in mean scores from baseline to end-of-treatment for IIEF Q3 (35.7 vs. 19.9%) and Q4 (68.4 vs. 26.5%) were significant in patients receiving sildenafil compared with those receiving placebo (P = 0.0001). Moreover, the percent of improved erections (GEQ, 66.6 vs. 28.6%) and successful intercourse attempts (63 vs. 33%) was significantly increased with sildenafil compared with placebo. Improvements in sexual function were seen irrespective of the degree of ED severity. Adverse events were generally mild to moderate in severity, with headache (20 vs. 8%), flushing (18 vs. 3%), and dyspepsia (8 vs. 1%) reported more often in the sildenafil than in placebo-treated patients. CONCLUSIONS: Treatment with sildenafil for ED was effective, resulting in an increased percentage of successful attempts at intercourse, and was well tolerated among men with type 1 diabetes. PMID- 12547850 TI - Plasma adiponectin plays an important role in improving insulin resistance with glimepiride in elderly type 2 diabetic subjects. AB - OBJECTIVE: We investigated the effect of glimepiride, a third-generation sulfonylurea hypoglycemic agent, on insulin resistance in elderly patients with type 2 diabetes, in connection with plasma adiponectin and 8-epi-prostagrandin F2alpha (8-epi-PGF2alpha), an oxidative stress marker. RESEARCH DESIGN AND METHODS: A total of 17 elderly patients with type 2 diabetes received 12 weeks of treatment with glimepiride. Homeostasis assessment model of insulin resistance (HOMA-IR), homeostasis assessment model of beta-cell function, HbA(1c), C-peptide in 24-h pooled urine (urine CPR), and plasma concentrations of 8-epi-PGF2alpha, tumor necrosis factor-alpha (TNF-alpha), plasminogen activator inhibitor type 1, and adiponectin were measured at various times. The metabolic clearance rate of glucose (MCR-g) was also assessed by a hyperinsulinemic-euglycemic clamp. RESULTS: After 8 weeks of glimepiride treatment, significant reductions were observed in HbA(1c) (from 8.4 +/- 1.9 to 6.9 +/- 1.0%), HOMA-IR (from 2.54 +/- 2.25 to 1.69 +/- 0.95%), and plasma TNF-alpha concentrations (from 4.0 +/- 2.0 to 2.6 +/- 2.5 pg/ml). MCR-g was significantly increased from 3.92 +/- 1.09 to 5.73 +/- 1.47 mg. kg(-1). min(-1). Plasma adiponectin increased from 6.61 +/- 3.06 to 10.2 +/- 7.14 micro g/ml. In control subjects, who maintained conventional treatment, no significant changes were observed in any of these markers. CONCLUSIONS: Glimepiride remarkably improved insulin resistance, suggested by a significant reduction in HOMA-IR, an increase in MCR-g, and a reduction in HbA(1c) without changing extrapancreatic beta-cell function and urine CPR. Increased plasma adiponectin and decreased plasma TNF-alpha may underlie the improvement of insulin resistance with glimepiride. PMID- 12547851 TI - Diagnostic strategies to detect glucose intolerance in a multiethnic population. AB - OBJECTIVE: Identifying individuals who have elevated glucose concentrations is important for clinicians so that preventive strategies can be invoked, and it is useful for researchers who study associations between elevated glucose and adverse health outcomes. These methods should be applicable worldwide across different ethnic groups. Therefore, the objective of our analysis was to determine whether using the fasting glucose and HbA(1c) together could improve the classification of individuals with impaired glucose tolerance and diabetes in a multiethnic cohort randomly assembled in Canada. RESEARCH DESIGN AND METHODS: We determined the optimum diagnostic criteria to identify people with abnormal glucose tolerance using fasting plasma glucose, 2-h post-glucose load plasma glucose, and HbA(1c) in 936 Canadians of South Asian, Chinese, and European descent. RESULTS: The sensitivity of the American Diabetes Association (ADA) criteria to diagnose diabetes compared with the World Health Organization definitions was poor at 48.3% (95% confidence interval [CI] 35.7-61.0). Using a receiver operator characteristic curve, the optimum combined cut-point using fasting glucose and HbA(1c) to diagnose diabetes was a fasting glucose > or =5.7 mmol/l and an HbA(1c) > or =5.9%. These cut-points were associated with a sensitivity and specificity of 71.7% (60.3-83.1) and 95.0% (93.5-96.4), respectively, a positive likelihood ratio (LR) of 14.3 (9.6-19.0), and a negative LR of 0.3 (0.2-0.4). Significant ethnic variation in the sensitivity and specificity of this approach was observed: 47.4% (24.9-69.8) and 97.6% (95.9 99.4) among Europeans, 78.6% (57.1-100) and 95.9% (93.6-98.2) among Chinese, and 85.2% (71.8-98.6) and 91.3% (88.1-94.6) among South Asians, respectively. Participants with impaired glucose tolerance could not be identified reliably using the fasting glucose or HbA(1c) alone or in combination. CONCLUSIONS: The sensitivity of the ADA criteria to diagnose diabetes is low, and there is substantial variation between ethnic groups. Fasting glucose and HbA(1c) may be used together to improve the identification of individuals who have diabetes, allowing clinicians to streamline the use of the oral glucose tolerance test. PMID- 12547852 TI - Target-seeking behavior of plasma glucose with exercise in type 1 diabetes. AB - OBJECTIVE: To investigate the reproducibility of the plasma glucose (PG) response to exercise in subjects with type 1 diabetes on a nonintensive insulin regimen. RESEARCH DESIGN AND METHODS: Subjects cycled for 45 min at 50% VO(2max) on two occasions (studies 1 and 2) either 1 h after lunch and usual insulin (protocol A) or after overnight fasting without morning insulin (protocol B). Identical diet, activity, and insulin (twice daily neutral and intermediate) were maintained before and during each study day. A total of 13 type 1 diabetic subjects (6 men and 7 women, BMI 24.0 +/- 0.9 kg/m(2) [means +/- SE], age 42.6 +/- 2.7 years, diabetes duration 14.1 +/- 2.8 years) completed protocol A, and 7 (3 men and 4 women, BMI 25.8 +/- 1.3 kg/m(2), age 39.7 +/- 1.3 years, diabetes duration 14 +/- 4.4 years) completed protocol B. RESULTS: In protocol A (fed), the fall in PG during exercise was 4.5 +/- 1.0 and 5.0 +/- 0.8 mmol/l in studies 1 and 2, respectively, whereas in protocol B (fasted), it was 0.6 +/- 0.8 and 3.4 +/- 1.6 mmol/l. Regression analysis of the change in PG in protocol A in study 1 versus study 2 showed poor reproducibility (r(2) = 0.12, P = 0.25) despite uniform conditions. In protocol B, the fall in PG was more reproducible (r(2) = 0.81, P = 0.006). In fed subjects, there was better (P = 0.01) and clinically useful reproducibility of the PG at exercise completion (r(2) = 0.77, P = 0.0001) compared with preexercise. CONCLUSIONS: These results indicate poor reproducibility of the change in PG during exercise after feeding in type 1 diabetes on nonintensive insulin regimens but reasonable reproducibility when fasting. Exercise apparently decreases the glycemic variability after feeding, so that PG concentrations after exercise seek a reproducible "target." Thus, the absolute PG level after a typical bout of exercise in the fed state should be a good guide to carbohydrate or insulin adjustment on subsequent occasions. PMID- 12547853 TI - Dietary fat intake as risk factor for the development of diabetes: multinational, multicenter study of the Mediterranean Group for the Study of Diabetes (MGSD). AB - OBJECTIVE: To investigate the role of dietary factors in the development of type 2 diabetes. RESEARCH DESIGN AND METHODS: In the context of the Multinational MGSD Nutrition Study, three groups of subjects were studied: 204 subjects with recently diagnosed diabetes (RDM), 42 subjects with undiagnosed diabetes (UDM) (American Diabetes Association criteria-fasting plasma glucose [FPG] > or =126 mg/dl), and 55 subjects with impaired fasting glucose (IFG) (FPG > or =110 and <126 mg/dl). Each group was compared with a control group of nondiabetic subjects, matched one by one for center, sex, age, and BMI. Nutritional habits were evaluated by a dietary history method, validated against the 3-day diet diary. In RDM, the questionnaire referred to the nutritional habits before the diagnosis of diabetes. Demographic data were collected, and anthropometrical and biochemical measurements were taken. RESULTS: Compared with control subjects, RDM more frequently had a family history of diabetes (49.0 vs. 14.2%; P < 0.001), exercised less (exercise index 53.5 vs. 64.4; P < 0.01), and more frequently had sedentary professions (47.5 vs. 27.4%; P < 0.001). Carbohydrates contributed less to their energy intake (53.5 vs. 55.1%; P < 0.05), whereas total fat (30.2 +/- 0.5 vs. 27.8 +/- 0.5%; P < 0.001) and animal fat (12.2 +/- 0.3 vs. 10.8 +/- 0.3%; P < 0.01) contributed more and the plant-to-animal fat ratio was lower (1.5 +/- 0.1 vs. 1.8 +/- 0.1; P < 0.01). UDM more frequently had a family history of diabetes (38.1 vs. 19.0%; P < 0.05) and sedentary professions (58.5 vs. 34.1%; P < 0.05), carbohydrates contributed less to their energy intake (47.6 +/- 1.7 vs. 52.8 +/- 1.4%; P < 0.05), total fat (34.7 +/- 1.5 vs. 30.4 +/- 1.2%; P < 0.05) and animal fat (14.2 +/- 0.9 vs. 10.6 +/- 0.7%; P < 0.05) contributed more, and the plant-to-animal fat ratio was lower (1.6 +/- 0.2 vs. 2.3 +/- 0.4; P < 0.05). IFG differed only in the prevalence of family history of diabetes (32.7 vs. 16.4%; P < 0.05). CONCLUSIONS: Our data support the view that increased animal fat intake is associated with the presence of diabetes. PMID- 12547854 TI - Epidemiology of diabetes among Arab Americans. AB - OBJECTIVE: To examine the prevalence of diabetes and glucose intolerance by age and sex in the Arab-American community of Dearborn, Michigan. RESEARCH DESIGN AND METHODS: Participants were randomly selected adult Arab Americans, 20-75 years of age, from randomly selected households in Dearborn, Michigan. Demographic and anthropometric data were recorded. Glucose tolerance was assessed with 2-h 75-g oral glucose tolerance tests and classified according to 1997 American Diabetes Association and 1998 World Health Organization criteria. RESULTS: A total of 626 eligible adults were selected, and 542 participated (87% response rate). Because prevalence increases with age and the overall response rate for women (328/352; 93%) was higher than that for men (214/274; 78%), prevalence rates were adjusted for age and sex. The overall prevalence of diabetes was 15.5% (95% CI 12.2-18.7%) in women and 20.1% (15.0-25.2%) in men (P = 0.13). The prevalence of previously diagnosed diabetes was similar to that of undiagnosed diabetes. Impaired glucose tolerance (IGT) and/or impaired fasting glucose (IFG) were present in 16.8% (12.8 20.8%) of women and 29.7% (23.4-35.9%) of men (P = 0.0007). The combined rates of glucose intolerance (diabetes, IGT, and IFG) were 32.3% (27.8-36.7%) for women and 49.8% (43.1-56.4%) for men (P < 0.0001). Among younger adults, the prevalence in men was higher than that in women. As expected, subjects with diabetes or IGT/IFG were older and had greater BMI and waist-to-hip ratios than subjects with normal glucose tolerance. CONCLUSIONS: The prevalence of diabetes and glucose intolerance is extremely high among adult Arab Americans in Michigan and represents a major clinical and public health problem. Community-based intervention programs to prevent and treat diabetes are urgently needed. PMID- 12547855 TI - Diabetes as a predictor of change in functional status among older Mexican Americans: a population-based cohort study. AB - OBJECTIVE: Epidemiological studies have demonstrated that older Mexican Americans are at high risk for type 2 diabetes and its complications. Type 2 diabetes leads to a more rapid decline in functional status among older Mexican Americans with diabetes. This study was designed to examine the impact of diabetes on change in self-reported functional status over a 2-year period among older Mexican Americans with diabetes. RESEARCH DESIGN AND METHODS: We performed a longitudinal analysis with repeated measurements of functional limitations in a cohort of Mexican Americans aged > or =60 years in the Sacramento Area Latino Study on Aging (SALSA). Diabetes was diagnosed on the basis of self-report of physician diagnosis, medication use, and fasting plasma glucose. Functional status was measured by assessment of activities of daily living (ADL) and instrumental activities of daily living (IADL) at baseline and 1 and 2 years. RESULTS: Of 1,789 SALSA participants, 585 (33%) had diabetes at baseline. Diabetic subjects reported 74% more limitations than nondiabetic subjects in ADL (summary score for number of limitations, 0.99 vs. 0.57; P = 0.002) and 50% more limitations in IADL (summary score for number of limitations, 7.83 vs. 5.25; P < 0.0001). The annual rate of increase in limitations of ADL and IADL was 0.046 and 0.033 (log scale) on each scale among diabetic subjects compared with 0.013 and 0.003 (log scale) among nondiabetic subjects (P < 0.0005). Complications of diabetes were found to increase ADL and IADL limitations among diabetic subjects. Longer duration of diabetes was also associated with an increase in ADL and IADL limitations. CONCLUSIONS: There was lower baseline functional status and a more rapid decline in functional status among older Mexican Americans with diabetes versus those without diabetes. PMID- 12547856 TI - Longitudinal studies of incidence and progression of diabetic retinopathy assessed by retinal photography in pima indians. AB - OBJECTIVE: To examine incidence and progression of retinopathy using retinal photographs in Pima Indians and to compare the results with those obtained when retinopathy is assessed by direct ophthalmoscopy. RESEARCH DESIGN AND METHODS: We undertook an analysis of examinations conducted between 1 April 1982 and 31 December 1990 in residents of the Gila River Community in central Arizona. Data were taken from 411 people with diabetes who had at least two examinations during this period. Incidence and progression of retinopathy were evaluated by retinal photography and fundoscopy, and hazard rate ratios were calculated for various potential risk factors measured at baseline. RESULTS: Previously diagnosed retinopathy tended to progress, except in individuals with minimal nonproliferative retinopathy, among whom follow-up examinations were more likely to show no retinopathy. Diabetes duration (hazard rate ratio [HRR], 1.06 per year difference [P = 0.007]), hyperglycemia (HbA(1) HRR, 1.27 per 1% difference [P < 0.0001]), the type of treatment for diabetes (insulin use HRR, 3.06 [P = 0.0007], and oral hypoglycemic use HRR, 2.40 [P = 0.0034], compared with individuals on no pharmacotherapy), and macroalbuminuria (HRR, 2.86, compared with individuals without macroalbuminuria [P = 0.0486]) were associated with the development of retinopathy. Although fundoscopy detected fewer cases of retinopathy, HRRs for most risk factors were similar when retinopathy was assessed by fundoscopy rather than retinal photography. CONCLUSIONS: Although retinopathy tends to worsen over time, some eyes show improvement, especially in patients with minimal nonproliferative retinopathy. As in other populations, glycemic control is the major modifiable risk factor for the development and progression of retinopathy. PMID- 12547857 TI - Outcome of pregnancy among immigrant women with diabetes. AB - OBJECTIVE: We studied outcome of pregnancy among immigrant women with diabetes. The women came from regions of the world with high incidence of impaired glucose tolerance and type 2 diabetes. Prevalences, secular trends, and sociodemographic risk factors of diabetes were also explored. RESEARCH DESIGN AND METHODS: Data from the Medical Birth Registry of Norway on all births from 1988 to 1998 for mothers born in South Asia and North Africa (11,268) and Norway (601,785) were analyzed. RESULTS: The prevalence of pregestational diabetes among the immigrants was 8.9/1,000 births, which was more than twice the rate among ethnic Norwegians (3.6/1,000). Time trends indicated increasing prevalences in both groups. Among the immigrants, diabetes was closely associated with maternal age. Maternal diabetes was associated with a significantly increased risk of pregnancy complications in both study groups. Increased risks were found for low birth weight, macrosomia, preterm birth, preeclampsia, and cesarean sections. Among ethnic Norwegians, maternal diabetes conferred a significantly increased risk of infant perinatal death (odds ratio 2.00, 95% CI 1,44-2.77). In the sample of immigrant women with predominantly type 2 diabetes, maternal diabetes was not significantly associated with perinatal death or congenital malformations in the offspring. CONCLUSIONS: The high prevalence of diabetes among immigrants from South Asia and North Africa represents a challenge for health care providers. To prevent adverse pregnancy outcomes and later cardiovascular and renal morbidity among these groups, early diagnosis of diabetes, adequate metabolic control, and relevant preventive measures are warranted. PMID- 12547858 TI - Identifying hepatic nuclear factor 1alpha mutations in children and young adults with a clinical diagnosis of type 1 diabetes. AB - OBJECTIVE: HNF-1alpha gene mutations (MODY3) present with marked hyperglycemia in lean young adults and may, therefore, be mistaken for type 1 diabetes, with implications for individual treatment and risk of diabetes in other family members. We examined the prevalence of HNF-1alpha mutations in families with three generations of diabetes identified in a population-based study of childhood diabetes, representing a subpopulation in which misclassification was likely. RESEARCH DESIGN AND METHODS: In a study population of 1,470 families, 36 families (2.4%) with three affected generations were identified. In the 18 families in whom DNA samples were available, islet autoantibody testing, HLA class II genotyping, and HNF-1alpha sequencing were performed. RESULTS: At least one islet autoantibody was found in 13 of 14 probands, and diabetes-associated HLA class II haplotypes were found in 17 of 18. One proband, who had no islet autoantibodies and was homozygous for the protective HLA haplotype DRB1*02-DQB1*0602, had a novel HNF-1alpha heterozygous nonsense mutation (R54X). This mutation cosegregated with diabetes in the family. The proband, his brother, mother, and maternal grandmother were diagnosed with type 1 diabetes aged 14-18 years and treated with insulin (0.39-0.74 units/kg) from diagnosis. The mother has since been successfully transferred to sulfonylurea treatment. CONCLUSIONS: Family history alone is of limited value in identification of individuals with HNF 1alpha mutations, and we propose a stepwise approach that restricts sequencing of the HNF-1alpha gene to those with a family history of diabetes who also test negative for islet autoantibodies. PMID- 12547859 TI - Urinary alpha1-microglobulin as a marker of nephropathy in type 2 diabetic Asian subjects in Singapore. AB - OBJECTIVE: This study examines urinary alpha(1)-microglobulin as a marker of early nephropathy in type 2 diabetic Chinese, Malays, and Asian Indians in Singapore. RESEARCH DESIGN AND METHODS: A cross-sectional study was performed on 590 consecutive type 2 diabetic patients (296 males, 294 females) who were on routine follow-up at a primary care clinic. Information was obtained from interviews, case notes, and blood and urine samples. Because the distribution of urinary alpha(1)-microglobulin levels was highly skewed, these levels were log transformed, and geometric means were calculated. There was correction for variability in urine flow by dividing by urine creatinine levels, given as mg/mmol urine creatinine, and adjustment for confounding variables. RESULTS: Urinary alpha(1)-microglobulin was higher in men than in women and was directly related to age, but no ethnic differences were apparent. It was directly related to duration of diabetes, with adjusted geometric means of 1.19 and 1.43 mg/mmol urine creatinine for a duration of <10 and > or =10 years, respectively (P = 0.07). Urinary alpha(1)-microglobulin was highest in patients on insulin, followed by those on oral medication and then those on diet alone (adjusted geometric means: 1.47, 1.36, and 0.86 mg/mmol urine creatinine, respectively; P = 0.01). Levels were also higher in patients with poor glucose control, as measured by HbA(1c), fasting plasma glucose, and 2-h postprandial plasma glucose (P < 0.01 for each). Urinary alpha(1)-microglobulin was directly related to albuminuria, with adjusted geometric means for normoalbuminuria, microalbuminuria, and macroalbuminuria of 1.06, 1.47, and 4.72 mg/mmol urine creatinine, respectively (P < 0.01). However, of patients with normoalbuminuria, 33.6% had raised urinary alpha(1)-microglobulin. Likewise, of patients with normal urinary alpha(1) microglobulin, 27.6% had albuminuria. CONCLUSIONS: Urinary alpha(1)-microglobulin was related to duration, severity, and control of diabetes. Urinary alpha(1) microglobulin and albumin were directly related, but in some patients, one was present in the absence of the other. Hence, in addition to albuminuria (which measures glomerular dysfunction), urinary alpha(1)-microglobulin (which measures proximal tubular dysfunction) is useful for the early detection of nephropathy in diabetic subjects. PMID- 12547860 TI - Low birth weight and high birth weight infants are both at an increased risk to have type 2 diabetes among schoolchildren in taiwan. AB - OBJECTIVE: To study the effect of birth weight on risk of type 2 diabetes in the schoolchildren in Taiwan. RESEARCH DESIGN AND METHODS: From 1992 to 1997, all schoolchildren aged 6-18 years were screened for diabetes in Taiwan Province. This cohort consisted of 1,966 patients with diabetes and 1,780 randomly selected subjects with normal fasting glycemia (NFG). Questionnaire interviewing was designed to classify diabetes. The birth weight was obtained from the Taiwan's Birth Registry. After merging the data, there were 978 subjects, including 429 with type 2 diabetes and 549 with of NFG enrolled in the present analyses. RESULTS: The odds ratios (95% CI) for type 2 diabetes, after adjusting age, sex, BMI, family history of diabetes, and socioeconomic status, were 2.91 (1.25-6.76) for children with low birth weight (<2,500 g) and 1.78 (1.04-3.06) for those with high birth weight (> or =4,000 g) when compared with the referent group (birth weight 3,000-3,499 g). The risk of diabetes was still 64% higher in the high birth weight group [odds ratio (OR) 1.64 (95% CI 0.91-2.96)], even after adjustment for gestational diabetes mellitus (GDM). Patients with type 2 diabetes who were born with high birth weight were more likely to have a higher BMI and diastolic blood pressure as well as a higher family history of diabetes compared with those with low birth weight. CONCLUSIONS: A U-shaped relationship between birth weight and risk of type 2 diabetes was found in the schoolchildren aged 6 18 years in Taiwan. Schoolchildren with type 2 diabetes who were born with low birth weight had different metabolic phenotypes compared with those born with high birth weight. PMID- 12547861 TI - The incidence of retinopathy 10 years after diagnosis in young adult people with diabetes: results from the nationwide population-based Diabetes Incidence Study in Sweden (DISS). AB - OBJECTIVE: To estimate the prevalence and severity of diabetic retinopathy (DR) 10 years after diagnosis in a nationwide population-based cohort study of young adult diabetic patients in Sweden. RESEARCH DESIGN AND METHODS: The Diabetes Incidence Study in Sweden (DISS) aims to register all incident cases of diabetes aged 15-34 years in Sweden. In 1987-1988, 806 cases were reported, and 627 (78%) of them were followed up with regard to retinopathy 8-10 years later. The assessment was based on retinal photographs in most cases (86%). RESULTS: Ten years after diagnosis, retinopathy was found in 247 patients (39%). The retinopathy was mild in 206 (33%), whereas 30 (4.8%) patients had moderate nonproliferative DR (NPDR) and 11 (1.8%) had proliferative DR (PDR). Patients with retinopathy had worse glycemic control during the years than patients without (HbA(1c) 8.1 +/- 1.5% and 6.8 +/- 1.2%, respectively; P < 0.001). In a Cox regression analysis, time to retinopathy was related to high HbA(1c) (P < 0.001) and high BMI (P = 0.001). Patients with type 2 diabetes had an increased prevalence of severe retinopathy (NPDR or PDR) compared with those with type 1 diabetes (14 of 93 [15%] versus no or mild 24 of 471 [5%], respectively; P < 0.001). CONCLUSIONS: Despite modern diabetes management, 39% of young adult diabetic patients developed retinopathy within the first 10 years of the disease. Nevertheless, compared with the prevalence of retinopathy (63%), after a similar duration of diabetes before the Diabetes Control and Complications Trial, this prevalence was clearly lower. Current treatment aimed to achieve strict glycemic control has reduced the risk for developing retinopathy. PMID- 12547862 TI - Hypertension management in patients with diabetes: the need for more aggressive therapy. AB - OBJECTIVE: Clinical trials have demonstrated the importance of tight blood pressure control among patients with diabetes. However, little is known regarding the management of hypertension in patients with coexisting diabetes. To examine this issue, we addressed 1) whether hypertensive patients with coexisting diabetes are achieving lower levels of blood pressure than patients without diabetes, 2) whether there are differences in the intensity of antihypertensive medication therapy provided to patients with and without diabetes, and 3) whether diabetes management affects decisions to increase antihypertensive medication therapy. RESEARCH DESIGN AND METHODS: We abstracted medical records to collect detailed information on 2 years of care provided for 800 male veterans with hypertension. We compared patients with and without diabetes on intensity of therapy and blood pressure control. Intensity of therapy was described using a previously validated measure that captures the likelihood of an increase in antihypertensive medications. We also determined whether increases in antihypertensive medications were less likely at those visits in which the diabetes medications were being adjusted. RESULTS: Of the 274 hypertensive patients with diabetes, 73% had a blood pressure > or =140/90 mmHg, compared with 66% in the 526 patients without diabetes (P = 0.04). Diabetic patients also received significantly (P = 0.05) less intensive antihypertensive medication therapy than patients without diabetes. Less intensive therapy in diabetic patients could not be explained by clinicians being distracted by the treatment for diabetes. CONCLUSIONS: There is an urgent need to improve hypertension care and blood pressure control in patients with diabetes. Additional information is required to understand why clinicians are not more aggressive in managing blood pressure when patients also have diabetes. PMID- 12547863 TI - The effects of diabetes on the risks of major cardiovascular diseases and death in the Asia-Pacific region. AB - OBJECTIVE: To provide reliable age- and region-specific estimates of the associations between diabetes and major cardiovascular diseases and death in populations from the Asia-Pacific region. RESEARCH DESIGN AND METHODS: Twenty four cohort studies from Asia, Australia, and New Zealand (median follow-up, 5.4 years) provided individual participant data from 161,214 people (58% from Asia) of whom 4,873 had a history of diabetes at baseline. The associations of diabetes with the risks of coronary heart disease, stroke, and cause-specific mortality during follow-up were estimated using time-dependent Cox models, stratified by study cohort and sex and adjusted for age at risk. RESULTS: In all, 9,277 deaths occurred (3,635 from cardiovascular disease). The hazard ratio (95% CI) associated with diabetes was 1.97 (1.72-2.25) for fatal cardiovascular disease; there were similar hazard ratios for fatal coronary heart disease, fatal stroke, and composites of fatal and nonfatal outcomes. For all cardiovascular outcomes, hazard ratios were similar in Asian and non-Asian populations and in men and women, but were greater in younger than older individuals. For noncardiovascular death, the hazard ratio was 1.56 (1.38-1.77), with separately significant increases in the risks of death from renal disease, cancer, respiratory infections, and other infective causes. The hazard ratio for all-causes mortality was 1.68 (1.55-1.84), with similar ratios in Asian and non-Asian populations, but with significantly higher ratios in younger than older individuals. CONCLUSIONS: The relative effect of diabetes on the risks of cardiovascular disease and death in Asian populations is much the same as that in the largely Caucasian populations of Australia and New Zealand. Hazard ratios were severalfold greater in younger people than older people. The rapidly growing prevalence of diabetes in Asia heralds a large increase in the incidence of diabetes-related death in the coming decades. PMID- 12547864 TI - Quality of care for patients diagnosed with diabetes at screening. AB - OBJECTIVE: Screening for diabetes has the potential to be an effective intervention, especially if patients have intensive treatment of their newly diagnosed diabetes and comorbid hypertension. We wished to determine the process and quality of diabetes care for patients diagnosed with diabetes by systematic screening. RESEARCH DESIGN AND METHODS: A total of 1,253 users of the Durham Veterans Affairs Medical Center aged 45-64 years who did not report having diabetes were screened for diabetes with an HbA(1c) test. All subjects with an HbA(1c) level > or =6.0% were invited for follow-up blood pressure and fasting plasma glucose (FPG) measurements. A case of unrecognized diabetes was defined as HbA(1c) > or =7.0% or FPG > or =126 mg/dl. For each of the 56 patients for whom we made a new diagnosis of diabetes, we notified the patient's primary care provider of this diagnosis. One year after diagnosis, we reviewed these patients' medical records for traditional diabetes performance measures as well as blood pressure. Follow-up blood pressure was also ascertained from medical record review for all subjects with HbA(1c) > or =6.0% who did not have diabetes. We compared blood pressure changes between patients with and without diabetes. RESULTS: Among patients diagnosed with diabetes at screening, 34 of 53 (64%) had evidence of diet or medical treatment for their diabetes, 42 of 53 (79%) had HbA(1c) measured within the year after diagnosis, 32 of 53 (60%) had cholesterol measured, 25 of 53 (47%) received foot examinations, 29 of 53 (55%) had eye examinations performed by an eye specialist, and 16 of 53 (30%) had any measure of urine protein. The mean blood pressure decline over the year after diagnosis for patients with diabetes was 2.3 mmHg; this decline was similar to that found for 183 patients in the study without diabetes (change in blood pressure, -3.6 mmHg). At baseline, 48% of patients with diabetes had blood pressure <140/90, compared with 40% of patients without diabetes; 1 year later, the same 48% of patients with diabetes had blood pressure <140/90, compared with 56% of patients without diabetes (P = 0.31 for comparing the change in percent in control between groups). CONCLUSIONS: Patients with diabetes diagnosed at screening achieve less tight blood pressure control than similar patients without diabetes. Primary care providers do not appear to manage diabetes diagnosed at screening as intensively as long-standing diabetes and do not improve the management of hypertension given the new diagnosis of diabetes. PMID- 12547865 TI - Association between regional adipose tissue distribution and both type 2 diabetes and impaired glucose tolerance in elderly men and women. AB - OBJECTIVE: We examined whether regional adipose tissue distribution, specifically that of skeletal muscle fat and visceral abdominal fat aggregation, is characteristic of elderly individuals with hyperinsulinemia, type 2 diabetes, and impaired glucose tolerance (IGT). RESEARCH DESIGN AND METHODS: A total of 2,964 elderly men and women (mean age 73.6 years) were recruited for cross-sectional comparisons of diabetes or glucose tolerance, generalized obesity with dual energy X-ray absorptiometry, and regional body fat distribution with computed tomography. RESULTS-Approximately one-third of men with type 2 diabetes and less than half of women with type 2 diabetes were obese (BMI > or =30 kg/m(2)). Despite similar amounts of subcutaneous thigh fat, intermuscular fat was higher in subjects with type 2 diabetes and IGT than in subjects with normal glucose tolerance (NGT) (11.2 +/- 9.4, 10.3 +/- 5.8, and 9.2 +/- 5.9 cm(2) for men; 12.1 +/- 6.1, 10.9 +/- 6.5, and 9.4 +/- 5.3 cm(2) for women; both P < 0.0001). Visceral abdominal fat was also higher in men and women with type 2 diabetes and IGT than in subjects with NGT (172 +/- 79, 163 +/- 72, and 145 +/- 66 cm(2) for men; 162 +/- 66, 141 +/- 60, and 116 +/- 54 cm(2) for women; both P < 0.0001 across groups). Higher rates of intermuscular fat and visceral abdominal fat were associated with higher fasting insulin in normal-weight (BMI <25 kg/m(2)) men (r = 0.24 for intermuscular fat, r = 0.37 for visceral abdominal fat, both P < 0.0001) and women (r = 0.20 for intermuscular fat, r = 0.40 for visceral abdominal fat, both P < 0.0001). These associations were not found in obese subjects. CONCLUSIONS: Elderly men and women with normal body weight may be at risk for metabolic abnormalities, including type 2 diabetes, if they possess an inordinate amount of muscle fat or visceral abdominal fat. PMID- 12547866 TI - A prospective study of self-reported sleep duration and incident diabetes in women. AB - Short-term sleep restriction results in impaired glucose tolerance. To test whether habitually short sleep duration increases the risk of developing diabetes, we studied a cohort of 70,026 women enrolled in the Nurses Health Study, without diabetes at baseline, and who responded to a question about daily sleep duration in 1986. Subjects were followed until 1996 for the diagnosis of diabetes (1,969 cases). Long and short sleep durations were associated with an increased risk of diabetes diagnosis. The relative risks (RRs) for short (slept < or =5 h per day) and long (slept > or =9 h per day) sleepers were 1.57 (95% CI 1.28-1.92) and 1.47 (1.19-1.80), respectively. After adjustment for BMI and a variety of confounders, the RR was not significantly increased for short sleepers (1.18 [0.96-1.44]) but remained modestly increased for long sleepers (1.29 [1.05 1.59]). We then performed a similar analysis using only symptomatic cases (n = 1,187). Adjusted RRs for symptomatic diabetes were modestly elevated in both short (1.34 [1.04-1.72]) and long (1.35 [1.04-1.75]) sleepers. Our data suggest that the association between a reduced self-reported sleep duration and diabetes diagnosis could be due to confounding by BMI, or sleep restriction may mediate its effects on diabetes through weight gain. Sleep restriction may be an independent risk factor for developing symptomatic diabetes. PMID- 12547867 TI - Gestational diabetes mellitus: is a diagnosis associated with an increase in maternal anxiety and stress in the short and intermediate term? AB - OBJECTIVE: To examine anxiety levels of women diagnosed with gestational diabetes mellitus (GDM) and to compare these with glucose-tolerant (GT) women at similar stages of pregnancy. RESEARCH DESIGN AND METHODS: Prospective longitudinal study conducted on 50 women with GDM and 50 GT women. All women completed the Mental Health Inventory (MHI-5) forms and the Speilberger State-Trait Anxiety Inventory (STAI) at the beginning of the third trimester, antepartum, and 6 weeks postpartum. Specific questions were also assessed using a Likert scale. RESULTS: Women with GDM, compared with GT women, had a higher level of anxiety (state rather than trait) at the time of the first assessment. However, before delivery and in the postpartum period, there were no significant differences in anxiety scores between the two groups. Women in both groups were positive about being tested for GDM and wished to be tested during future pregnancies. CONCLUSIONS: There were no sustained increased levels of anxiety for women diagnosed with GDM. Concerns expressed about causing sustained maternal anxiety by testing for GDM could not be substantiated. PMID- 12547868 TI - Short-term, delayed, and working memory are impaired during hypoglycemia in individuals with type 1 diabetes. AB - OBJECTIVE: To examine the effects of acute insulin-induced hypoglycemia on short term, delayed, and working memory in individuals with type 1 diabetes. RESEARCH DESIGN AND METHODS: A hyperinsulinemic glucose clamp was used to maintain arterialized blood glucose level at either 4.5 mmol/l (euglycemia) or 2.5 mmol/l (hypoglycemia) on two separate occasions in 16 adults with type 1 diabetes. The participants completed tests of immediate and delayed verbal memory, immediate and delayed visual memory, and working memory during each experimental condition. Two other mental tests, the Trail Making B Test and the Digit Symbol Test, were also administered. RESULTS: Performance in tests of immediate verbal and immediate visual memory was significantly impaired during hypoglycemia. The effect of hypoglycemia on working memory and delayed memory was more profound. Performance in the nonmemory tests, the Trail Making B Test, and the Digit Symbol Test also deteriorated during hypoglycemia. CONCLUSIONS: All of the memory systems examined in the present study were affected significantly by acute hypoglycemia, particularly working memory and delayed memory. Mild (self-treated) hypoglycemia is common in individuals with insulin-treated diabetes; therefore, these observed effects of hypoglycemia on memory are of potential clinical importance because they could interfere with many everyday activities. PMID- 12547869 TI - Self-reported diabetes and health behaviors in remote indigenous communities in northern queensland, australia. AB - OBJECTIVE: This study examines associations between self-reported diabetes and self-reported smoking, alcohol consumption, fruit consumption, and participation in adequate exercise in remote indigenous communities, using data from the Well Persons' Health Check (WPHC). RESEARCH DESIGN AND METHODS: The WPHC was a cross sectional survey of 2,862 indigenous individuals (1,602 Aborigines, 1,074 Torres Strait Islanders, and 186 persons of joint descent) aged > or =15 years. The study was conducted in 26 remote communities in northern Queensland, Australia, between March 1998 and October 2000. RESULTS: A total of 32% of individuals with self-reported diabetes and 25% of other individuals reported eating enough fruit, according to National Health and Medical Research Council criteria: odds ratio (OR) 1.407 (95% CI 1.108-1.786), P = 0.006. After adjustment for age, sex, and ethnicity, no significant difference could be observed: adjusted OR 1.22 (0.944 1.574), P = 0.128. A total of 58% of participants who reported diabetes and 51% of others reported adequate exercise: OR 0.761 (0.609-0.952), P = 0.018. This difference was not significant after adjustment for age, sex, and ethnicity: adjusted OR 0.896 (0.705-1.14), P = 0.370. A total of 43% of individuals who reported diabetes and 72% of others reported consuming alcohol: OR 0.295 (0.235 0.369), P < 0.001. After adjustment for age, sex, and ethnicity, this difference was still significant: adjusted OR 0.550 (0.428-0.709), P < 0.001. Diabetic drinkers consumed alcohol at harmful levels similar to those of nondiabetic drinkers (P = 0.691). A total of 40% of individuals who reported diabetes and 63% of other persons were tobacco smokers: OR 0.403 (0.322-0.505), P < 0.001. Although this crude difference was attenuated by adjustment for age, sex, and ethnicity, persons with self-reported diabetes were still significantly less likely to smoke tobacco than other participants: adjusted OR 0.666 (0.521-0.852), P = 0.001. Smoking prevalence among the diabetic indigenous participants was more than double that in nondiabetic nonindigenous Australians. CONCLUSIONS: This study suggests that indigenous individuals with diabetes living in rural and remote communities are not adopting lifestyle changes required for optimal self management of the disease. This contributes to the large excess of mortality and morbidity experienced by this population. PMID- 12547870 TI - Validation of a counseling strategy to promote the adoption and the maintenance of physical activity by type 2 diabetic subjects. AB - OBJECTIVE: There is enough evidence that physical activity is an effective therapeutic tool in the management of type 2 diabetes. The present study was designed to validate a counseling strategy that could be used by physicians in their daily outpatient practice to promote the adoption and maintenance of physical activity by type 2 diabetic subjects. RESEARCH DESIGN AND METHODS: The long-term (2-year) efficacy of the behavioral approach (n = 182) was compared with usual care treatment (n = 158) in two matched, randomized groups of patients with type 2 diabetes who had been referred to our Outpatient Diabetes Center. The outcome of the intervention was consistent patient achievement of an energy expenditure of >10 metabolic equivalents (METs)-h/week through voluntary physical activity. RESULTS: After 2 years, 69% of the patients in the intervention group (27.1 +/- 2.0 METs x h/week) and 18% of the control group (4.1 +/- 0.8 METs x h/week) achieved the target (P < 0.001) with significant (P < 0.001) improvements in BMI (intervention group 28.9 +/- 0.2 versus control group 30.4 +/- 0.3 kg/m(2)) and HbA(1c) (intervention group 7.0 +/- 0.1 versus control group 7.6 +/- 0.1%). CONCLUSIONS: This randomized, controlled study shows that physicians can motivate most patients with type 2 diabetes to exercise long-term and emphasizes the value of individual behavioral approaches in daily practice. PMID- 12547871 TI - Prevalence and predictors of sexual dysfunction in patients with type 1 diabetes. AB - OBJECTIVE: This study aimed to 1) measure the prevalence of sexual dysfunction in patients with diabetes; 2) describe how descriptive variables, psychological variables, diabetic complications, and sexual dysfunction relate in patients with diabetes; and 3) describe the predictors of sexual dysfunction in patients with diabetes. RESEARCH DESIGN AND METHODS: A total of 240 adult type 1 diabetic patients visiting the outpatient diabetes clinic of a university hospital completed questionnaires evaluating psychological adjustment to diabetes and sexual functioning. Medical records were used to obtain HbA(1c) values as well as information on microvascular diabetic complications. RESULTS: Sexual dysfunction was reported by 27% of women and 22% of men. No differences were found between sexes in type of reported sexual dysfunction. In men, but not in women, sexual dysfunction was related to age, BMI, duration of diabetes, and diabetic complications. No correlation with HbA(1c) was found in either sex. In women, but not in men, sexual dysfunction was related to depression and the quality of the partner relationship. Binary logistic regression demonstrated that, in men, the significant predictors of sexual dysfunction were higher age and presence of complications, whereas, in women, sexual dysfunction was related to depression. CONCLUSIONS: Both women and men with diabetes are at increased risk for sexual dysfunction. This study suggests that in men with diabetes, sexual dysfunction is related to somatic and psychological factors, whereas in women with diabetes, psychological factors are more predominant. PMID- 12547872 TI - Prevalence and costs of major depression among elderly claimants with diabetes. AB - OBJECTIVE: To compare the odds of major depression among Medicare claimants with and without diabetes and to test whether annual medical payments are greater for those with both diabetes and major depression than for those with diabetes alone. RESEARCH DESIGN AND METHODS: This retrospective analysis relies on claims data from the 1997 Medicare 5% Standard Analytic Files. Using these data, we statistically determined whether the odds of major depression are greater among elderly claimants with diabetes after controlling for age, race/ethnicity, and sex. We then used regression analysis on a sample of over 220,000 elderly claimants with diabetes to test whether payments for non-mental health-related services are greater for those with both diabetes and major depression (n = 4,203) than for those with diabetes alone. RESULTS: Our findings indicate that the odds of major depression are significantly greater among elderly Medicare claimants with diabetes than among those without diabetes (OR 1.58 +/- 0.05). We also found that elderly claimants with both diabetes and major depression seek treatment for more services and spend more time in inpatient facilities, and as a result incur higher medical costs than claimants with diabetes but without major depression. These results hold even after excluding services related to mental health treatment. CONCLUSIONS: This analysis suggests that treatment for major depression among claimants with diabetes may reduce total medical costs if treatment results in a decrease in utilization for general medical services in the future. PMID- 12547873 TI - Control of postprandial plasma glucose by an oral insulin product (HIM2) in patients with type 2 diabetes. AB - OBJECTIVE: The objectives of this exploratory study were to assess the postprandial glucose-lowering effects and evaluate the safety and tolerability of single, escalating doses of an oral insulin product, hexyl-insulin monoconjugate 2 (HIM2), in patients with type 2 diabetes. Subcutaneous insulin and oral placebo were also administered for comparison. RESEARCH DESIGN AND METHODS: Eighteen patients with type 2 diabetes were enrolled in this randomized, single-blind, placebo-controlled, three-way crossover, dose-escalation study. A single dose of each of the following study drugs was administered to each patient on 3 separate days: oral HIM2 (at one of three dose levels: 0.375, 0.5, or 1.0 mg/kg), subcutaneous regular insulin (8 units Humulin R), and oral placebo. At 30 min after dosing, patients ingested a standardized test meal (16 oz/720 calories of Boost Plus). Serial blood samples were collected for determination of plasma glucose and insulin concentrations during the 4-h postdose period. RESULTS: The mean glucose area under the curve for 0 to 240 min (AUC(0-240)) values were lower following administration of 0.5 and 1.0 mg/kg HIM2 vs. placebo (1,097.1 vs. 1,196.9 and 801.1 vs. 992.1 mg x h(-1) x dl(-1), respectively). This difference was statistically significant at the 1.0-mg/kg HIM2 dose level. Insulin exposure, as measured by insulin AUC(0-240) values, for the 0.375-, 0.5-, and 1.0-mg/kg dose levels of HIM2 were 169.9, 193.1, and 230.8 micro U x h(-1) x ml(-1), respectively; insulin AUC(0-240) values for placebo were 165.8, 196.1, and 169.2 micro U x h(-1) x ml(-1), respectively. The mean glucose AUC(0-240) values were similar following administration of 0.5 and 1.0 mg/kg HIM2 vs. subcutaneous insulin (1,097.1 vs. 1,048.0 and 801.1 vs. 875.2 mg x h(-1) x dl(-1), respectively). For pooled data from the 0.5- and 1.0-mg/kg dose groups, the HIM2/subcutaneous insulin ratios for the 2-h postprandial glucose concentration (0.97, 95% CI 0.90-1.06), maximum postprandial glucose concentration (0.99, 95% CI 0.93-1.06), and glucose AUC(0-240) (0.98, 95% CI 0.9-1.06) were within 10% of unity, implying glucodynamic equivalence. Although HIM2 (0.5 and 1.0 mg/kg) and subcutaneous insulin (8 units) provided comparable control of postprandial plasma glucose concentrations, HIM2 resulted in peripheral insulin concentrations that were lower than subcutaneous insulin (mean insulin AUC(0-240) of 193.1 vs. 233.6 and 230.8 vs. 270.3 micro U x h(-1) x ml(-1), respectively). CONCLUSIONS: Single, oral doses of HIM2 were safe and well tolerated. HIM2 (0.5 and 1.0 mg/kg) was more effective than placebo and as effective as subcutaneous regular insulin (8 units) at controlling postprandial glycemia with respect to the following parameters: 2-h postprandial glucose concentration, maximum glucose concentration, and glucose AUC(0-240). This occurred even though peripheral insulin concentrations were lower following the administration of HIM2 (0.5 and 1.0 mg/kg) than subcutaneous insulin. Thus, HIM2 therapy may control postprandial glycemia without causing peripheral hyperinsulinemia in patients with type 2 diabetes. PMID- 12547874 TI - Atorvastatin dose-dependently decreases hepatic lipase activity in type 2 diabetes: effect of sex and the LIPC promoter variant. AB - OBJECTIVE: Hepatic lipase (HL) is involved in the metabolism of several lipoproteins and may contribute to the atherogenic lipid profile in type 2 diabetes. Little is known about the effect of cholesterol synthesis inhibitors on HL activity in relation to sex and the hepatic lipase gene, the LIPC promoter variant in type 2 diabetes. Therefore, we studied the effect of atorvastatin 10 mg (A10) and 80 mg (A80) on HL activity in 198 patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: Patients (aged 45-75 years, without manifest coronary artery disease, total cholesterol 4.0-8.0 mmol/l, and fasting triglycerides [TG] 1.5-6.0 mmol/l) were included in a double-blind, randomized, placebo-controlled trial for 30 weeks (Diabetes Atorvastatin Lipid Intervention study). RESULTS: HL activity at baseline was significantly higher in our population compared with an age-matched control group without type 2 diabetes (406 +/- 150 vs. 357 +/- 118 units/l). HL activity in men versus women (443 +/- 158 vs. 358 +/- 127 units/l), in carriers of the LIPC C/C allele versus carriers of the T/T allele (444 +/- 142 vs. 227 +/- 96 units/l), and in Caucasians versus blacks (415 +/- 150 vs. 260 +/- 127 units/l) all differed significantly (P < 0.001). Atorvastatin dose-dependently decreased HL (A10, -11%; A80, -22%; both P < 0.001). Neither sex nor the LIPC C-->T variation influenced the effect of atorvastatin on HL activity. CONCLUSIONS: Sex, LIPC promoter variant, and ethnicity significantly contribute to the baseline variance in HL activity. Atorvastatin treatment in diabetic dyslipidemia results in a significant dose dependent decrease in HL activity, regardless of sex or the LIPC promoter variant. PMID- 12547875 TI - Delineation of prevalence and risk factors for early coronary artery disease by electron beam computed tomography in young adults with type 1 diabetes. AB - OBJECTIVE: Type 1 diabetes increases the risk for coronary artery disease (CAD), but limited information is available regarding the early natural history of this process. Electron beam tomography (EBT) can measure coronary artery calcification (CAC), an early marker for CAD. This study was designed to assess the prevalence and risk factors for CAC in young adults with established type 1 diabetes. RESEARCH DESIGN AND METHODS: A total of 101 subjects aged 17-28 years with type 1 diabetes of over 5 years' duration and no history of heart disease underwent cardiac EBT with calcium scoring. Medical histories were obtained and physical examinations were conducted to document the presence of cardiac risk factors as well as evidence of microvasculopathy and diabetic arthropathy. Laboratory evaluation included measurement of fasting lipoproteins, homocysteine concentration, lipoprotein(a) [Lp(a)], urinary microalbumin, and HbA(1c). Contingency table analysis was used to assess bivariate relationships. Logistic regression was employed to construct a parsimonious model of independent risk factors. RESULTS: Eleven subjects (10.9%) had CAC. Smokers were nearly five times more likely than nonsmokers to have CAC (P = 0.03). In addition, each 0.36-mm/l increment of Lp(a) was associated with a 10% increased risk for CAC (P = 0.05) after controlling for potentially confounding factors. There was no association of other CAD or diabetes risk factors studied with CAC. CONCLUSIONS: The prevalence of early CAD as evidenced by CAC in young adults with type 1 diabetes is significant. Smoking and Lp(a) levels independently predict the presence of CAC. Additional study is necessary to delineate the natural history of CAC and the role of risk factor modification to prevent progression of CAD in this high risk population. PMID- 12547876 TI - Pulse wave velocity as an indicator of atherosclerosis in impaired fasting glucose: the Tanno and Sobetsu study. AB - OBJECTIVE: Brachial-ankle pulse wave velocity (baPWV), as an indicator of atherosclerosis in impaired fasting glucose (IFG), was studied in 232 subjects randomly selected from inhabitants of two rural communities in Japan. RESEARCH DESIGN AND METHODS: BMI, systolic blood pressure (SBP), fasting blood glucose (FBS), lipid parameters, ankle brachial pressure index (ABI), and baPWV were measured in each subject. ABI and baPWV were measured using the recently developed device, form ABI/PWV. The subjects were divided into three groups according FBS level: a normal group consisting of subjects with FBS <110 mg/dl, an IFG group consisting of subjects with FBS 110-125 mg/dl, and a diabetic group consisting of subjects with FBS > or =126 mg/dl and subjects taking hypoglycemic agents. The parameters in the three groups were compared. RESULTS-It was found that the baPWV value increased with increasing plasma glucose level. Significant differences were found between the baPWV values in the normal and IFG groups (1,518 vs. 1,673 cm/s, P = 0.01) and in the normal and diabetic groups (1,518 vs. 1,771 cm/s, P < 0.0001). The results of multiple regression analysis showed that FBS was closely related to baPWV as well as to age and SBP. CONCLUSIONS: The relationship between IFG and atherosclerosis remains controversial. Further studies are needed to evaluate whether strict control of blood glucose level in patients with IFG will result in the prevention of atherosclerosis progression. PMID- 12547877 TI - The influence of shoe design on plantar pressures in neuropathic feet. AB - OBJECTIVE: The aim of this study was to investigate the effect of shoe design on the plantar pressure dynamics of patients with diabetic neuropathy during walking. RESEARCH DESIGN AND METHODS: Three shoe design categories were tested. Total contact area and biomechanical variables in multiple areas under the foot were measured. RESULTS: Shoes with a rocker bottom principle reduced pressure 35 65% underneath the heel and the central metatarsal heads. Increased contact area did not result in significant pressure reductions underneath the forefoot. Pressure dynamics underneath the heel and medial forefoot (first metatarsal head and hallux) on average showed no significant differences among the different shoes with a cushioning insole. CONCLUSIONS: The most effective way to offload the forefoot of patients with neuropathic feet is through the use of the rocker sole principle. In general, the effect of an insole depends on the design characteristics of a shoe. Predicting the effect of therapeutic footwear on an individual scale, however, remains difficult. Therefore, in-shoe pressure measurements seem to be necessary to evaluate a therapeutic shoe prescription in certain individual cases. PMID- 12547878 TI - Maggot therapy for treating diabetic foot ulcers unresponsive to conventional therapy. AB - OBJECTIVE: To assess the efficacy of maggot therapy for treating foot and leg ulcers in diabetic patients failing conventional therapy. RESEARCH DESIGN AND METHODS: Retrospective comparison of changes in necrotic and total surface area of chronic wounds treated with either maggot therapy or standard (control) surgical or nonsurgical therapy. RESULTS: In this cohort of 18 patients with 20 nonhealing ulcers, six wounds were treated with conventional therapy, six with maggot therapy, and eight with conventional therapy first, then maggot therapy. Repeated measures ANOVA indicated no significant change in necrotic tissue, except when factoring for treatment (F [1.7, 34] = 5.27, P = 0.013). During the first 14 days of conventional therapy, there was no significant debridement of necrotic tissue; during the same period with maggot therapy, necrotic tissue decreased by an average of 4.1 cm(2) (P = 0.02). After 5 weeks of therapy, conventionally treated wounds were still covered with necrotic tissue over 33% of their surface, whereas after only 4 weeks of therapy maggot-treated wounds were completely debrided (P = 0.001). Maggot therapy was also associated with hastened growth of granulation tissue and greater wound healing rates. CONCLUSIONS: Maggot therapy was more effective and efficient in debriding nonhealing foot and leg ulcers in male diabetic veterans than was continued conventional care. PMID- 12547879 TI - Similar genetic features and different islet cell autoantibody pattern of latent autoimmune diabetes in adults (LADA) compared with adult-onset type 1 diabetes with rapid progression. AB - OBJECTIVE: To compare the clinical parameters, C-peptide levels, pattern of islet cell-specific autoantibodies, and prevalence of predisposing genotypes in subjects with latent autoimmune diabetes in adults (LADA) and those with adult onset type 1 diabetes with rapid progression. RESEARCH DESIGN AND METHODS: We evaluated the clinical parameters, C-peptide levels, and islet cell-specific autoantibodies in 54 LADA, 57 adult-onset type 1 diabetic, and 190 type 2 diabetic patients. Islet cell autoantibodies were also compared between subgroups of newly diagnosed patients with LADA and those with newly diagnosed adult-onset and childhood-onset type 1 diabetes. The genetic study was performed in subjects with LADA and those with adult-onset type 1 diabetes in comparison with a control population. RESULTS: There were no differences in the clinical parameters between LADA and adult-onset type 1 diabetes. Patients with LADA had lower BMI (P < 0.0001), waist-to-hip ratio (0.0029), total cholesterol (P = 0.001), and triglycerides (P = 0.001); higher HDL cholesterol levels (P < 0.0001); and lower prevalence of hypertension (P = 0.0028) compared with patients with type 2 diabetes. C-peptide levels were similar at onset (P = 0.403) but decreased less rapidly in LADA than in adult-onset type 1 diabetes (P = 0.0253). Single autoantibody positivity was more often seen in LADA than in type 1 diabetes (P = 0.0001). The prevalence of predisposing HLA-DQB1*0302, -DR4, -DR3, and -DR3/DR4 genotypes and the DR4-DQB1*0302 haplotype were increased in both LADA and adult onset type 1 diabetic subjects compared with the control population. There were no differences in the frequencies of these risk alleles and haplotypes between the two patient groups. CONCLUSIONS: Subjects with LADA had clinical characteristics similar to those with adult-onset type 1 diabetes with rapid progression. C-peptide levels did not differ at onset but decreased less rapidly in LADA. Patients with LADA rather had single islet cell-specific autoantibody positivity. The prevalence of HLA-DQB1*0302, -DR4, -DR3, and -DR3/DR4 risk alleles and the DR4-DQB1*0302 high-risk haplotype did not differ in the two forms of autoimmune diabetes. PMID- 12547880 TI - Association of NAD(P)H oxidase p22 phox gene variation with advanced carotid atherosclerosis in Japanese type 2 diabetes. AB - OBJECTIVE: To evaluate the association between the C242T polymorphism of the p22 phox gene, an essential component of NAD(P)H oxidase in the vasculature, with intima-media thickness (IMT) of the carotid artery and risk factors for atherosclerosis in type 2 diabetic subjects. RESEARCH DESIGN AND METHODS: C242T polymorphism of the p22 phox gene was detected by polymerase chain reaction restriction fragment-length polymorphism in 200 Japanese type 2 diabetic subjects and 215 nondiabetic subjects. We examined the association with this mutation and carotid atherosclerosis as well as the patients' clinical characteristics and the level of 8-hydroxy-2'deoxyguanosine (8-OHdG) as an index of oxidative DNA damage. RESULTS: The diabetic subjects with the TC+TT genotypes displayed a significantly lower average IMT (1.13 +/- 0.31 vs. 1.31 +/- 0.34 mm; P = 0.0099) and a not significantly lower serum 8-OHdG level than those with the CC genotype, despite no difference in the risk factors. Stepwise multiple regression analysis showed that the risk factors for increased IMT in the diabetic subjects were systolic blood pressure (P = 0.0042) and p22 phox CC genotype (P = 0.0151). In nondiabetic subjects, the average IMT of the TC+TT group was not different from that of the CC group (0.85 +/- 0.14 vs. 0.94 +/- 0.30 mm, P = 0.417). Fasting plasma insulin concentration (41.4 +/- 15.6 vs. 64.2 +/- 59.4 pmol/l, P = 0.0098) and insulin resistance index of homeostasis model assessment (HOMA-R) (1.58 +/- 0.66 vs. 2.60 +/- 2.56, P = 0.0066) were significantly lower in the TC+TT group than in the CC group. CONCLUSIONS: These results show that the C242T mutation in the p22 phox gene is associated with progression of asymptomatic atherosclerosis in the subjects with type 2 diabetes and is also associated with insulin resistance in nondiabetic subjects. PMID- 12547882 TI - Urinary cadmium, impaired fasting glucose, and diabetes in the NHANES III. AB - OBJECTIVE: Increasing rates of type 2 diabetes worldwide suggest that diabetes may be caused by environmental toxins. Cadmium is a widespread environmental pollutant that accumulates in the pancreas and exerts diabetogenic effects in animals. To test the hypothesis that exposure to cadmium is associated with impaired fasting glucose and type 2 diabetes, we examined the associations between urinary cadmium and the prevalence of impaired fasting glucose (prediabetes) and diabetes in the Third National Health and Nutrition Examination Survey (NHANES III). RESEARCH DESIGN AND METHODS: We analyzed data on 8,722 adults > or =40 years of age from the NHANES III (1988-1994), a cross-sectional health survey of a nationally representative sample of the noninstitutionalized civilian U.S. population. We studied urinary levels of cadmium (adjusted for urine creatinine) in relation to the prevalence of impaired fasting glucose and diabetes, using the criteria of the American Diabetes Association. RESULTS: After adjustment for age, ethnicity, sex, and BMI, the odds of impaired fasting glucose and diabetes increased dose-dependently with elevations in urinary cadmium from 0 0.99 to 1.00-1.99 and > or =2 micro g/g creatinine (impaired fasting glucose, odds ratio [OR] 1.48, 95% CI 1.21-1.82 and OR 2.05, 95% CI 1.42-2.95; diabetes, OR 1.24, 95% CI 1.06-1.45 and OR 1.45, 95% CI 1.07-1.97). CONCLUSIONS: In this large cross-sectional study, urinary cadmium levels are significantly and dose dependently associated with both impaired fasting glucose and diabetes. These findings, which require confirmation in prospective studies, suggest that cadmium may cause prediabetes and diabetes in humans. PMID- 12547881 TI - The K121Q polymorphism of the PC-1 gene is associated with insulin resistance but not with dyslipidemia. AB - OBJECTIVE: To investigate the relationship of the K121Q polymorphism of the plasma cell glycoprotein 1 (PC-1) gene with insulin resistance, insulin secretion, and lipids and lipoproteins. RESEARCH DESIGN AND METHODS: Altogether, 110 normoglycemic subjects (group I) underwent a hyperinsulinemic-euglycemic clamp for evaluation of insulin sensitivity. The first-phase insulin secretion was determined by the intravenous glucose tolerance test (IVGTT) in a separate sample of 295 normoglycemic subjects (group II). RESULTS: The 121Q allele (genotypes K121Q and Q121Q) compared with the K121K genotype was related to higher fasting insulin levels (group I: 69.6 +/- 45.6 vs. 51.9 +/- 28.4 pmol/l [mean +/- SD], P = 0.050; group II: 66.6 +/- 38.8 vs. 53.8 +/- 26.6 pmol/l, P = 0.009). In group I, subjects carrying the 121Q allele compared with subjects with the K121K genotype had lower rates of whole-body glucose uptake (51.17 +/- 12.07 vs. 60.12 +/- 14.86 micro mol x kg(-1) x min(-1), P = 0.012) and nonoxidative glucose disposal (33.71 +/- 10.51 vs. 41.51 +/- 13.36 micro mol x kg(-1) x min( 1), P = 0.015) during the clamp. In group II, there was no significant difference between the 121Q allele carriers and subjects with the K121K genotype in total first-phase insulin secretion during the first 10 min of the IVGTT (2,973 +/- 2,224 vs. 2,520 +/- 1,492 pmol. l(-1). min(-1), P = 0.415). No association of the K121Q polymorphism with serum lipids and lipoproteins was found. CONCLUSIONS: In healthy normoglycemic Finnish subjects, the K121Q polymorphism of the PC-1 gene is associated with insulin resistance but not with impaired insulin secretion or dyslipidemia. PMID- 12547883 TI - Insulin sensitivity and lipid metabolism in human CD36 deficiency. AB - OBJECTIVE: CD36 has been proposed as a fatty acid translocase and a receptor for HDL and oxidized LDL. The association between CD36 deficiency and insulin resistance remains controversial. We investigated glucose and lipid metabolism in human CD36 deficiency. RESEARCH DESIGN AND METHODS: A total of 61 type I CD36 deficient patients and 25 control subjects were examined. Diabetes was defined as fasting glucose level > or =7 mmol/l or use of hypoglycemic agents. A homeostasis model assessment (HOMA) index was evaluated in patients without diabetes. Insulin resistance was defined as a HOMA index > or =1.73 (sensitivity 64.3%, specificity 78.9%; J Japan Diab Soc, 2000). RESULTS: Diabetes was identified in 12 (20%) of the 61 CD36-deficient patients. Fasting glucose, HbA(1c), and total cholesterol levels in the diabetic CD36-deficient patients were significantly higher than in the control subjects and the nondiabetic CD36-deficient patients. Regardless of diabetes, HDL cholesterol concentrations in the CD36-deficient patients were significantly higher than in the control subjects. The nondiabetic CD36-deficient patients had higher triglyceride concentrations than the control subjects, and triglyceride concentrations were higher in the diabetic CD36-deficient patients than in the nondiabetic CD36-deficient patients. The prevalence of insulin resistance in the nondiabetic CD36-deficient patients was similar to that in the control subjects. CONCLUSIONS: Human CD36 deficiency is not necessarily responsible for insulin resistance. Lipid abnormalities in CD36 deficiency may partly depend on the presence of diabetes, and increased levels of triglyceride and HDL cholesterol may be due to impaired binding of fatty acids and HDL to CD36 and subsequent clearance. PMID- 12547884 TI - Enhancement of endothelial function by pregnancy: inadequate response in women with type 1 diabetes. AB - OBJECTIVE: Pregnant women with type 1 diabetes have a substantially increased risk of vascular complications. Our aim was to study vascular function and metabolic and inflammatory risk factors during the antenatal and postpartum periods in women with type 1 diabetes compared with healthy control subjects. RESEARCH DESIGN AND METHODS: A total of 15 women with diabetes and 30 control subjects were recruited in the third trimester of pregnancy. Of these women, 9 case subjects and 16 control subjects were reinvestigated in the postnatal period. Blood samples were collected and microvascular skin perfusion was assessed in vivo using laser Doppler imaging and iontophoretic administration of endothelial-dependent (acetylcholine [ACH]) and endothelial-independent (sodium nitroprusside [SNP]) vasodilators. RESULTS: Microvascular responses in both control subjects (ACH, P = 0.018; SNP, P = 0.01) and diabetic women (ACH, P = 0.029; SNP, P = 0.105) were better during pregnancy than in the postnatal period, although responses in women with diabetes were significantly inferior to those in control subjects during both periods (all P < 0.001, two-way ANOVA). This dysfunction existed despite similar lipoprotein profiles. The difference in vascular responsivity between case and control subjects was significantly attenuated by adjustment for differences in HbA(1c) but not C-reactive protein concentrations in the two groups. CONCLUSIONS: Pregnancy enhances microvascular function, but in women with diabetes, such improvements are insufficient to attain responses seen in healthy nonpregnant women. This suggests a persistent vascular defect in young women with type 1 diabetes that may contribute to adverse pregnancy outcome. Our data suggest a role for the chronic effects of hyperglycemia in the impaired vascular responsiveness in such women. PMID- 12547885 TI - Aging per se does not influence glucose homeostasis: in vivo and in vitro evidence. AB - OBJECTIVE: To assess the effect of age on glucose metabolism by examining 1) glucose metabolism in young and middle-aged subjects when total or regional adiposity is taken into account and 2) in vitro glucose transport in adipose tissue explants from young and middle-aged women paired for total and abdominal adiposity. RESEARCH DESIGN AND METHODS: Study 1: body composition, subcutaneous abdominal and visceral adipose tissue areas, and fasting and oral glucose stimulated glucose and insulin were measured in 84 young and 81 middle-aged men and in 110 young and 91 middle-aged women. Study 2: glucose uptake in subcutaneous abdominal and visceral adipose tissue explants were measured in eight young and eight middle-aged women. RESULTS: Study 1: young and middle-aged men showed similar subcutaneous abdominal tissue area, whereas fat mass and visceral adipose tissue were greater in middle-aged than in young men (P < 0.01). Fat mass and subcutaneous and visceral adipose tissue areas were greater in middle-aged as compared with young women (P < 0.01). Fasting plasma glucose and the glucose response to an oral glucose tolerance test were significantly higher in middle-aged than in young men and women (P < 0.001). Statistical control for visceral adipose tissue area eliminated the difference seen in glucose response in men and women. Study 2: glucose transport in subcutaneous and omental adipose tissue did not differ between young and middle-aged women. CONCLUSIONS: 1) Visceral obesity, more than age per se, correlates with glucose intolerance in middle-aged subjects; 2) aging does not influence in vitro adipose tissue glucose uptake. PMID- 12547886 TI - Macrovascular risk and diagnostic criteria for type 2 diabetes: implications for the use of FPG and HbA(1c) for cost-effective screening. AB - OBJECTIVE: The use of fasting plasma glucose (FPG) level > or =7.0 mmol/l leads to underdiagnosis of type 2 diabetes compared with the oral glucose tolerance test (OGTT). The OGTT is of limited use for population screening. Most of the increase in cardiovascular risk in relation to increasing blood glucose occurs before the threshold at which the diagnosis of type 2 diabetes is made. The aim of this study was to evaluate the use of HbA(1c) and FPG as predictors of type 2 diabetes and cardiovascular risk and, accordingly, to develop a rational approach to screening for abnormalities of glucose tolerance. RESEARCH DESIGN AND METHODS: OGTT and measurement of HbA(1c) and FPG levels were performed in 505 subjects screened for type 2 diabetes. Anthropomorphic measurements were obtained. A cardiovascular risk factor questionnaire was completed. RESULTS: The subjects were aged 19-88 years (mean 53.8). The incidence of type 2 diabetes was 10.4% based on the OGTT and 4% based on an FPG level > or =7.0 mmol/l. Using high performance liquid chromatography (HPLC), HbA(1c) of <4.7 and > or =6.2% predicted with certainty the absence or presence of type 2 diabetes as defined by the OGTT. The corresponding cutoffs were <5.0 and > or =6.8% for HbA(1c) (DCA2000 HPLC device; Bayer Diagnostics, Mulgrave, Australia) and <4.7 and > or =6.4 mmol/l for FPG. However, 75-85% of subjects in each case had intermediate values, which were therefore nondiagnostic. Cardiovascular risk increased at least 2.2 times at an HbA(1c) level > or =6.2% (by HPLC), 1.8-2.2 times at an HbA(1c) level of 5.6-6.1% (by HPLC), 2 times at an FPG level > or =6.4 mmol/l, and 1.7-1.9 times at an FPG level of 5.6-6.3 mmol/l. CONCLUSIONS: Measurement of FPG and HbA(1c) levels will diagnose or exclude type 2 diabetes with certainty in a minority (15%) of people. There is a continuous relationship between FPG and HbA(1c) and cardiovascular risk. Accordingly, we propose that there is a rational basis for using either FPG and HbA(1c) for purposes of screening and assigning risk. Individuals with an HbA(1c) level of 5.6-6.1% and an FPG level of 5.6-6.3 mmol/l are at greatest risk for cardiovascular disease and should be targeted for further evaluation. An algorithm outlining a cost-effective approach is presented. PMID- 12547887 TI - Amputation and mortality in new-onset diabetic foot ulcers stratified by etiology. AB - OBJECTIVE: Foot ulcers and their complications are an important cause of morbidity and mortality in diabetes. The present study aims to examine the long term outcome in terms of amputations and mortality in patients with new-onset diabetic foot ulcers in subgroups stratified by etiology. RESEARCH DESIGN AND METHODS: Patients presenting with new ulcers (duration <1 month) to a dedicated diabetic foot clinic between 1994 and 1998 were studied. Outcomes were determined until March 2000 (or death) from podiatry, hospital, and district registers. Baseline clinical examination was done to classify ulcers as neuropathic, ischemic, or neuroischemic. Five-year amputation and mortality rates were derived from Kaplan-Meier survival analysis curves. RESULTS: Of the 185 patients studied, 41% had peripheral vascular disease (PVD) and 61% had neuropathy; 45%, 16%, and 24% of patients had neuropathic, ischemic, and neuroischemic ulcers, respectively. The mean follow-up period was 34 months (range 1-65) including survivors and patients who died during the study period. Five-year amputation rates were higher for ischemic (29%) and neuroischemic (25%) than neuropathic (11%) ulcers. Five-year mortality was 45%, 18%, and 55% for neuropathic, neuroischemic, and ischemic ulcers, respectively. Mortality was higher in ischemic ulcers than neuropathic ulcers. On multivariate regression analysis, only increasing age predicted shorter survival time. CONCLUSIONS: All types of diabetic foot ulcers are associated with high morbidity and mortality. The increased mortality appears to be independent of factors increasing ulcer risk that is, neuropathy and PVD-in patients with established foot ulcers. PMID- 12547888 TI - Effects of ethnicity and nephropathy on lower-extremity amputation risk among diabetic veterans. AB - OBJECTIVE: To describe ethnic differences in the risk of amputation in diabetic patients with diabetic nephropathy. RESEARCH DESIGN AND METHODS: A retrospective cohort study was conducted on a national cohort of diabetic patients who received primary care within the Veterans Affairs (VA) Health Care System. Hospitalizations for lower-limb amputations were established by ICD-9-CM procedure codes. Relative risk of amputation in diabetic patients with and without diabetic nephropathy was determined using Cox proportional hazard modeling for unadjusted and adjusted models. RESULTS: Of the 429,918 subjects identified with diabetes (mean age 64 +/- 11 years, 97.4% male), 3,289 individuals were determined to have had a lower-limb amputation during the study period. Compared with diabetic patients without amputations, amputees were on average older, more likely to belong to a minority group, and were more likely to have received treatment for more comorbid conditions. Asians were more likely to have toe amputations compared with whites or other ethnicities, while Native Americans were more likely to have below-the-knee amputations. Native Americans had the highest risk of amputation (RR 1.74, 95% CI 1.39-2.18), followed by African Americans (RR 1.41, 95% CI 1.34-1.48) and Hispanics (RR 1.28, 95% CI 1.20 1.38) compared with whites. The presence of diabetic nephropathy increased the risk of amputation threefold in all groups. Asian subjects with diabetes had the lowest adjusted relative risk of amputation (RR 0.31, 95% CI 0.19-0.50). CONCLUSIONS: Among diabetic patients, certain ethnic minority individuals have an increased risk of lower-extremity amputation compared with whites. Presence of diabetic nephropathy further increases this risk. PMID- 12547889 TI - Renal and metabolic effects of insulin lispro in type 2 diabetic subjects with overt nephropathy. AB - OBJECTIVE: To assess whether the insulin analog lispro may antagonize the renal effects of IGF-1, a mediator of glomerular hyperfiltration involved in the progression of diabetic and nondiabetic chronic nephropathies. RESEARCH DESIGN AND METHODS: In a randomized crossover study, we compared the renal and metabolic responses to regular or lispro insulin (0.1 units/kg body wt) administered after a euglycemic clamp and 5 and 30 min before a standard meal to 11 type 2 diabetic patients with macroalbuminuria. RESULTS: Two- and four-hour postprandial changes (vs. preprandial euglycemia) in glomerular filtration rate (GFR) followed a significantly different trend (5.8 +/- 5.0 vs. -6.3 +/- 4.7, P < 0.05; and 11.0 +/- 6.8 vs. 0.7 +/- 5.1%, P < 0.05) after regular insulin and lispro injection, respectively. After lispro, postprandial GFR changes were negatively correlated (r = -0.48, P = 0.0001) with plasma insulin concentration. After regular insulin, renal plasma flow increased in parallel with a decrease in renal vascular resistances. Both changes were fully prevented by lispro. Postprandial blood glucose maximum concentration (278 +/- 16 vs. 240 +/- 16 mg/dl, P < 0.01) and area under the curve (79,381 +/- 19,237 vs. 72,810 +/- 16,211 mg/dl per min, P < 0,05) were significantly lower after insulin lispro than after regular insulin injection, respectively, despite comparable postprandial insulin profiles. Changes in total and gluconeogenic amino acids followed a similar trend. Changes in blood glucose and plasma amino acids did not correlate with concomitant changes in GFR. CONCLUSIONS: In overt nephropathy of type 2 diabetes, lispro prevents glomerular hyperfiltration and offsets the renal effects of meal or meal associated hyperglycemia by mechanisms possibly related to IGF-1 antagonism. PMID- 12547890 TI - Quantifying the risk of infectious diseases for people with diabetes. AB - OBJECTIVE: In vitro evidence shows that immune function is compromised in people with diabetes. Although certain rare infections are more common and infection related mortality is higher, the risk of acquiring an infectious disease for diabetic patients has never been quantified. RESEARCH DESIGN AND METHODS: A retrospective cohort study using administrative data compared all people with diabetes in Ontario, Canada, on 1 April 1999 to matched nondiabetic people (n = 513,749 in each group). The risk ratios of having an infectious disease and of death attributable to infectious disease between those with and without diabetes were calculated. Secondary analysis individually examined common infectious diseases. The study was repeated using a second pair of cohorts defined in 1996 to confirm stability of the estimates. RESULTS: Nearly half of all people with diabetes had at least one hospitalization or physician claim for an infectious disease in each cohort year. The risk ratio for diabetic versus nondiabetic people was 1.21 (99% CI 1.20-1.22) in both cohort years. The risk ratio for infectious disease-related hospitalization was up to 2.17 (99% CI 2.10-2.23). The risk ratio for death attributable to infection was up to 1.92 (1.79-2.05). Many individual infections were more common in people with diabetes, especially serious bacterial infections. CONCLUSIONS: Diabetes confers an increased risk of developing and dying from an infectious disease, corroborating both in vitro evidence and commonly held clinical belief. In addition to microvascular and macrovascular sequelae, clinicians should consider infection a complication of diabetes. PMID- 12547891 TI - Barriers and strategies for research in Arab Americans. PMID- 12547892 TI - The potential therapeutic role of insulin in acute myocardial infarction in patients admitted to intensive care and in those with unspecified hyperglycemia. PMID- 12547893 TI - A tide in the affairs of medicine. PMID- 12547894 TI - Pharmacologic treatment of type 2 diabetes. PMID- 12547895 TI - Control of hypertension in diabetes. PMID- 12547896 TI - What's in a name: latent autoimmune diabetes of adults, type 1.5, adult-onset, and type 1 diabetes. PMID- 12547897 TI - Age and glucose intolerance: effect of fitness and fatness. PMID- 12547898 TI - Point: Diabetic Patients and Coronary Calcium: Risk stratification, compliance, and plaque progression. PMID- 12547899 TI - Counterpoint: Do people with diabetes benefit from coronary calcium scans? PMID- 12547900 TI - Progression of the carotid artery intima-media thickness in young patients with type 1 diabetes. PMID- 12547901 TI - Inflammatory parameters are independent predictors of severe epicardial coronary stenosis in asymptomatic diabetic patients with silent myocardial ischemia. PMID- 12547902 TI - Spending in the U.S. on advertising for fast foods, sodas, and automobiles: food for thought regarding the type 2 diabetes epidemic. PMID- 12547903 TI - MTHFR gene polymorphism as a risk factor for diabetic retinopathy in type 2 diabetic patients without serum creatinine elevation. PMID- 12547904 TI - Leptin is reduced in lean subjects with type 2 diabetes in bangladesh. PMID- 12547905 TI - A low glycemic diet significantly improves the 24-h blood glucose profile in people with type 2 diabetes, as assessed using the continuous glucose MiniMed monitor. PMID- 12547906 TI - Diabetes and cholesterol metabolism: the succinate hypothesis. PMID- 12547908 TI - The maltase-glucoamylase gene: common ancestry to sucrase-isomaltase with complementary starch digestion activities. AB - Brush-border maltase-glucoamylase (MGA) activity serves as the final step of small intestinal digestion of linear regions of dietary starch to glucose. Brush border sucrase-isomaltase (SI) activity is complementary, through digestion of branched starch linkages. Here we report the cloning and sequencing of human MGA gene and demonstrate its close evolutionary relationship to SI. The gene is approximately 82,000 bp long and located at chromosome 7q34. Forty-eight exons were identified. The 5' gene product, when expressed as the N-terminal protein sequence, hydrolyzes maltose and starch, but not sucrose, and is thus distinct from SI. The catalytic residue was identified by mutation of an aspartic acid and was found to be identical with that described for SI. The exon structures of MGA and SI were identical. This homology of genomic structure is even more impressive than the previously reported 59% amino acid sequence identity. The shared exon structures and peptide domains, including proton donors, suggest that MGA and SI evolved by duplication of an ancestral gene, which itself had already undergone tandem gene duplication. The complementary human enzyme activities allow digestion of the starches of plant origin that make up two-thirds of most diets. PMID- 12547909 TI - Hox gene clusters in the Indonesian coelacanth, Latimeria menadoensis. AB - The Hox genes encode transcription factors that play a key role in specifying body plans of metazoans. They are organized into clusters that contain up to 13 paralogue group members. The complex morphology of vertebrates has been attributed to the duplication of Hox clusters during vertebrate evolution. In contrast to the single Hox cluster in the amphioxus (Branchiostoma floridae), an invertebrate-chordate, mammals have four clusters containing 39 Hox genes. Ray finned fishes (Actinopterygii) such as zebrafish and fugu possess more than four Hox clusters. The coelacanth occupies a basal phylogenetic position among lobe finned fishes (Sarcopterygii), which gave rise to the tetrapod lineage. The lobe fins of sarcopterygians are considered to be the evolutionary precursors of tetrapod limbs. Thus, the characterization of Hox genes in the coelacanth should provide insights into the origin of tetrapod limbs. We have cloned the complete second exon of 33 Hox genes from the Indonesian coelacanth, Latimeria menadoensis, by extensive PCR survey and genome walking. Phylogenetic analysis shows that 32 of these genes have orthologs in the four mammalian HOX clusters, including three genes (HoxA6, D1, and D8) that are absent in ray-finned fishes. The remaining coelacanth gene is an ortholog of hoxc1 found in zebrafish but absent in mammals. Our results suggest that coelacanths have four Hox clusters bearing a gene complement more similar to mammals than to ray-finned fishes, but with an additional gene, HoxC1, which has been lost during the evolution of mammals from lobe-finned fishes. PMID- 12547910 TI - An evolutionary scenario for the transition to undifferentiated multicellularity. AB - The evolutionary transition from single cells toward multicellular forms of life represents one of the major transitions in the evolution of complex organisms. In this transition, single autonomously reproducing cells became parts of larger reproducing entities that eventually constituted a new unit of selection. The first step in the evolutionary transition to multicellularity likely was the evolution of simple, undifferentiated cell clusters. However, what the selective advantage of such cell clusters may have been remains unclear. Here, we argue that in populations of unicellular organisms with cooperative behavior, clustering may be beneficial by reducing interactions with noncooperative individuals. In support of this hypothesis, we present a set of computer simulations showing that clustering can evolve as a biological, heritable trait for cells that cooperate in the use of external energy resources. Following the evolution of simple cell clusters, further benefits could have arisen from the exchange of resources between cells of a cluster. PMID- 12547911 TI - Quantitative imaging of the T cell antitumor response by positron-emission tomography. AB - We describe a noninvasive, quantitative, and tomographic method to visualize lymphocytes within the whole animal. We used positron-emission tomography (PET) to follow the localization of adoptively transferred immune T lymphocytes. Splenic T cells from animals that had rejected a Moloney murine sarcoma virus/Moloney murine leukemia virus (M-MSV/M-MuLV)-induced tumor were marked with a PET reporter gene, injected into tumor-bearing mice, and imaged in a microPET by using a substrate specific for the reporter. Specific localization of immune T cells to the antigen-positive tumor was detected over time, by sequential imaging of the same animals. Naive T cells did not localize to the tumor site, indicating that preimmunization was required. Autoradiography and immunohistochemistry analysis corroborated the microPET data. The method we have developed can be used to assess the effects of immunomodulatory agents intended to potentiate the immune response to cancer, and can also be useful for the study of other cell mediated immune responses, including autoimmunity. PMID- 12547912 TI - Abrogation of postentry restriction of HIV-1-based lentiviral vector transduction in simian cells. AB - HIV-1 replication in simian cells is restricted at an early postentry step because of the presence of an inhibitory cellular factor. This block reduces the usefulness of HIV-1-based lentiviral vectors in primate animal models. Here, we demonstrate that substitution of the cyclophilin A (CyPA) binding region in the capsid of an HIV-1-based lentiviral vector (LV) with that of the macrophage tropic HIV-1 Ba-L resulted in a vector that was resistant to the inhibitory effect and efficiently transduced simian cells. Notably, the chimeric gag LV efficiently transduced primary simian hematopoietic progenitor cells, a critical cellular target in gene therapy. The alterations in the CyPA binding region did not affect CyPA incorporation; however, transduction by the gag chimeric LV seemed to be relatively insensitive to cyclosporin A, indicating that it does not require CyPA for early postentry steps. In dual infection experiments, the gag chimeric LV failed to remove the block to transduction of the WT LV, suggesting that the gag chimeric LV did not saturate the inhibitory simian cellular factor. These data suggest that the CyPA binding region of capsid contains a viral determinant involved in the postentry restriction of HIV-1-based lentiviral vectors. Overall, the findings demonstrate that the host range of HIV-1-based LV can be altered by modifications in the packaging construct. PMID- 12547913 TI - Mechanistic studies of beta-arylsulfotransferase IV. AB - Sulfotransferases are an important class of enzymes that catalyze the transfer of a sulfuryl group to a hydroxyl or amine moiety on various molecules including small-molecule drugs, steroids, hormones, carbohydrates, and proteins. They have been implicated in a number of disease states but remain poorly understood, complicating the design of specific, small-molecule inhibitors. A linear free energy analysis in both the forward and reverse directions indicates that the transfer of a sulfuryl group to an aryl hydroxyl group catalyzed by beta arylsulfotransferase IV likely proceeds by a dissociative (sulfotrioxide-like) mechanism. Values for the Bronsted coefficients (beta(nuc) and beta(lg)) are +0.33 and -0.45, giving Leffler alpha values of 0.19 and 0.61 for the forward and reverse reactions, respectively. PMID- 12547914 TI - GABAergic synapses made by a retinal dopaminergic neuron. AB - In the retina, dopaminergic amacrine (interplexiform) cells establish multiple synapses on the perikarya of AII amacrines, the neurons that distribute rod signals to on- and off-cone bipolars. We used triple-label immunocytochemistry and confocal microscopy to identify the receptors contained within the postsynaptic active zone of these synapses in both mouse and rat retinas. We found that at the interface between the dendrites of the dopaminergic neurons and the AII amacrine cell perikarya clusters of postsynaptic gamma-aminobutyric acid type A (GABA(A)) receptors are situated in register with aggregates of presynaptic organelles immunoreactive for GABA, the GABA vesicular transporter, and the vesicular monoamine transporter-2. D1 and D23 dopamine receptors, on the other hand, do not form clusters on the surface of the perikarya of AII amacrine cells. We suggest that the synapses between retinal dopaminergic neurons and AII amacrine cells are GABAergic and that both GABA and dopamine are released by the presynaptic endings. GABA acts on the ionotropic receptors clustered at the postsynaptic active zone, whereas dopamine diffuses to more distant, slower acting metabotropic receptors. PMID- 12547915 TI - Ecological community description using the food web, species abundance, and body size. AB - Measuring the numerical abundance and average body size of individuals of each species in an ecological community's food web reveals new patterns and illuminates old ones. This approach is illustrated using data from the pelagic community of a small lake: Tuesday Lake, Michigan, United States. Body mass varies almost 12 orders of magnitude. Numerical abundance varies almost 10 orders of magnitude. Biomass abundance (average body mass times numerical abundance) varies only 5 orders of magnitude. A new food web graph, which plots species and trophic links in the plane spanned by body mass and numerical abundance, illustrates the nearly inverse relationship between body mass and numerical abundance, as well as the pattern of energy flow in the community. Species with small average body mass occur low in the food web of Tuesday Lake and are numerically abundant. Larger-bodied species occur higher in the food web and are numerically rarer. Average body size explains more of the variation in numerical abundance than does trophic height. The trivariate description of an ecological community by using the food web, average body sizes, and numerical abundance includes many well studied bivariate and univariate relationships based on subsets of these three variables. We are not aware of any single community for which all of these relationships have been analyzed simultaneously. Our approach demonstrates the connectedness of ecological patterns traditionally treated as independent. Moreover, knowing the food web gives new insight into the disputed form of the allometric relationship between body mass and abundance. PMID- 12547916 TI - Chromatin insulation by a transcriptional activator. AB - In eukaryotic genomes, transcriptionally active regions are interspersed with silent chromatin that may repress genes in its vicinity. Chromatin insulators are elements that can shield a locus from repressive effects of flanking chromatin. Few such elements have been characterized in higher eukaryotes, but transcriptional activating elements are an invariant feature of active loci and have been shown to suppress transgene silencing. Hence, we have assessed the ability of a transcriptional activator to cause chromatin insulation, i.e., to relieve position effects at transgene integration sites in cultured cells. The transgene contained a series of binding sites for the metal-inducible transcriptional activator MTF, linked to a GFP reporter. Clones carrying single integrated transgenes were derived without selection for expression, and in most clones the transgene was silent. Induction of MTF resulted in transition of the transgene from the silent to the active state, prolongation of the active state, and a marked narrowing of the range of expression levels at different genomic sites. At one genomic site, prolonged induction of MTF resulted in suppression of transgene silencing that persisted after withdrawal of the induction stimulus. These results are consistent with MTF acting as a chromatin insulator and imply that transcriptional activating elements can insulate active loci against chromatin repression. PMID- 12547918 TI - Conservation genetics of the Far Eastern leopard (Panthera pardus orientalis). AB - The Far Eastern or Amur leopard (Panthera pardus orientalis) survives today as a tiny relict population of 25-40 individuals in the Russian Far East. The population descends from a 19th-century northeastern Asian subspecies whose range extended over southeastern Russia, the Korean peninsula, and northeastern China. A molecular genetic survey of nuclear microsatellite and mitochondrial DNA (mtDNA) sequence variation validates subspecies distinctiveness but also reveals a markedly reduced level of genetic variation. The amount of genetic diversity measured is the lowest among leopard subspecies and is comparable to the genetically depleted Florida panther and Asiatic lion populations. When considered in the context of nonphysiological perils that threaten small populations (e.g., chance mortality, poaching, climatic extremes, and infectious disease), the genetic and demographic data indicate a critically diminished wild population under severe threat of extinction. An established captive population of P. p. orientalis displays much higher diversity than the wild population sample, but nearly all captive individuals are derived from a history of genetic admixture with the adjacent Chinese subspecies, P. p. japonensis. The conservation management implications of potential restoration/augmentation of the wild population with immigrants from the captive population are discussed. PMID- 12547917 TI - A role for Mints in transmitter release: Mint 1 knockout mice exhibit impaired GABAergic synaptic transmission. AB - Mints (also called X11-like proteins) are adaptor proteins composed of divergent N-terminal sequences that bind to synaptic proteins such as CASK (Mint 1 only) and Munc18-1 (Mints 1 and 2) and conserved C-terminal PTB- and PDZ-domains that bind to widely distributed proteins such as APP, presenilins, and Ca(2+) channels (all Mints). We find that Mints 1 and 2 are similarly expressed in most neurons except for inhibitory interneurons that contain selectively high levels of Mint 1. Using knockout mice, we show that deletion of Mint 1 does not impair survival or alter the overall brain architecture, arguing against an essential developmental function of the Mint 1-CASK complex. In electrophysiological recordings in the hippocampus, we observed no changes in short- or long-term synaptic plasticity in excitatory synapses from Mint 1-deficient mice and detected no alterations in the ratio of alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) to N-methyl-d-aspartate (NMDA) receptor-mediated synaptic currents. Thus the Mint 1-CASK complex is not required for AMPA- and NMDA-receptor functions or for synaptic plasticity in excitatory synapses. In inhibitory synapses, however, we uncovered an approximately 3-fold increase in presynaptic paired-pulse depression, suggesting that deletion of Mint 1 impairs the regulation of gamma-aminobutyric acid release. Our data indicate that Mints 1 and 2 perform redundant synaptic functions that become apparent in Mint 1 deficient mice in inhibitory interneurons because these neurons selectively express higher levels of Mint 1 than Mint 2. PMID- 12547919 TI - Molecular identification of evolutionarily significant units in the Amazon River dolphin Inia sp. (Cetacea: Iniidae). AB - The Amazon river dolphin, genus Inia, is endemic to the major river basins of northern South America. No previous studies have focused on the genetic structure of this genus. In this work, 96 DNA samples from specimens of this genus were collected in the Orinoco basin (four rivers), the Putumayo River, a tributary of the Colombian Amazon and the Mamore, and the Tijamuchi and Ipurupuru rivers in theBolivian Amazon. These samples were used to amplify a fragment of 400 bp of the mitochondrial DNA (mtDNA) control region. In addition, 38 of these samples were also used to sequence 600 bp of the mitochondrial cytochrome b gene. The analysis of the population structure subdivision with an analysis of molecular variance (AMOVA) revealed important aspects about the genetic structure of Inia groups fromthese three geographically separate regions. By comparing the control region DNA and cytochrome b sequences, distinct types of nonshared haplotypes were observed. The net genetic divergence of control region sequences was 6.53% between the Orinoco and Bolivian rivers, 5.32% between the Putumayo and Bolivian rivers, and 2.50% between the Orinoco and Putumayo rivers. For the cytochrome b gene, these values were 2.48%, 2.98%, and 0.06%, respectively. The nucleotide sequences were analyzed phylogenetically using several genetic distance matrices and applying neighbor-joining, maximum likelihood, and maximum parsimony procedures. The results support the proposal to subdivide the Inia genus into at least two evolutionarily significant units: one confined to the Bolivian river basin and the other widely distributed across the Amazon and Orinoco basins. PMID- 12547920 TI - African-derived mitochondria in South American native cattle breeds (Bos taurus): evidence of a new taurine mitochondrial lineage. AB - This article reports the nucleotide diversity within the control region of 42 mitochondrial chromosomes belonging to five South American native cattle breeds (Bos taurus). Analysis of these data in conjunction with B. taurus and B. indicus sequences from Africa, Europe, the Near East, India, and Japan allowed the recognition of eight new mitochondrial haplotypes and their relative positions in a phylogenetic network. The structure of genetic variation among different hypothetical groupings was tested through the molecular variance decomposition, which was best explained by haplotype group components. Haplotypes surveyed were classified as European-related and African-related. Unexpectedly, two haplotypes within the African cluster were more divergent from the African consensus than the latter from the European consensus. A neighbor-joining tree shows the position of two haplotypes compared to European/African mitochondrial lineage splitting. This different and putatively ancestral mitochondrial lineage (AA) is supported by the calibration of sequence divergence based on the Bos-Bison separation. The European/African mitochondria divergence might be subsequent (67,100 years before present) to that between AA and Africans (84,700 years before present), also preceding domestication times. These genetic data could reflect the haplotype distribution of Iberian cattle five centuries ago. PMID- 12547921 TI - Geographic structure in the European flat oyster (Ostrea edulis L.) as revealed by Microsatellite polymorphism. AB - Genetic differentiation of the flat oyster (Ostrea edulis) was studied along the European coast, from Norway to the Black Sea, by means of variation at five microsatellite loci. The results show a mild but significant isolation-by distance profile, a noticeable between-sample variance in expected heterozygosity, and a tendency for Atlantic populations to be less variable than Mediterranean ones. This does not provide support for the existence of a single large panmictic population for this larvae-broadcasting species, but rather for the relative independence of local stocks. Comparison with data on allozyme variation from the literature confirms this view. It also leads us to suggest that the behavior of some sampled protein loci may depart from the average, so caution should be used when inferring neutral gene flow. PMID- 12547922 TI - Evolutionary trends of different repetitive DNA sequences during speciation in the genus secale. AB - The presence and distribution of two simple sequence repeats (SSRs), three highly repetitive sequences from rye, and the 5S rDNA have been investigated in 3 rye cultivars and 10 wild-related species of the genus SECALE: The following conclusions can be drawn in addition to detailed knowledge of the sequence content of chromatin in each accession studied: (1) Every species is unique in either or both the complement and chromosomal distribution of the six repeated sequences analyzed. (2) These sequences reveal multiple landmarks along all the rye chromosomes arms. (3) High polymorphism as well as heterozygosity between homologues in the distribution of the (AAG)(5) and (AAC)(5) was revealed in the outbreeding species of the Secale strictum complex. (4) It is possible to deduce trends in the complexity of repetitive DNA during the evolution of the genus. A possible evolutionary pathway that accounts for the present-day Secale species is presented. PMID- 12547923 TI - Expression of the narrow leaflet gene for yield and agronomic traits in soybean. AB - Genes that affect plant form and function may be used to enhance the yield of soybean [Glycine max (L.) Merr.]. Most soybean cultivars have broad (ovate) leaflets. A single gene, ln, controls inheritance for the narrow leaflet characteristic. Narrow leaflet cultivars (ln/ln) also tend to have a higher percentage of four-seeded pods than ovate (Ln/Ln) leaflet cultivars. Heterozygous (Ln/ln) plants have a leaflet shape intermediate between narrow and ovate. Determining the agronomic effects of the narrow leaflet allele (ln) in the heterozygous (Ln/ln) condition in soybean may have applications in practical plant breeding. We studied an ovate leaflet and a narrow leaflet cultivar, crosses between them in the F(1) and F(2), and backcrosses to both cultivars. The ratio of leaflet width to leaflet length accurately distinguished among narrow, ovate, and intermediate leaflet plants in the F(2) and backcross generations. In the F(2) generation, differences occurred among plants with different leaflet morphology. Narrow leaflet plants produced more seeds per pod and lower seed weight than ovate leaflet plants. Narrow and ovate leaflet plants produced comparable numbers of pods per plant and plant yield. Compared to ovate leaflet plants, intermediate leaflet plants produced similar numbers of seeds per pod and seed weight. Intermediate leaflet plants produced significantly more pods per plant and plant yield than plants with either ovate or narrow leaflets. The heterozygous condition at the locus for leaflet morphology resulted in heterosis for plant yield and may be of benefit in association with commercialization and development of hybrid soybean. PMID- 12547925 TI - Identification of a third fuzzless seed locus in upland cotton (Gossypium hirsutum L.). AB - Segregating populations were developed to evaluate the inheritance of the fuzzless seed phenotypes in upland cotton (Gossypium hirsutum L.). Accession 143 of the Mississippi Obsolete Variety Collection (MOVC) has a fuzzless seed phenotype. This line carries the n(2) locus which is recessive to the seed fuzz phenotype. Data from the F(2), BC(1)F(1), F(2:3), and BC(1)F(2) populations of DP 5690 x 143 fit a two-loci model for expression of the recessive fuzzless seed phenotype. Fuzzless seeds were obtained in n(2)n(2) plants when a second recessive locus (n(3)) was present. The dominant N(3) allele found in DP 5690 confers the fuzzy seed phenotype in homozygous n(2) plants. Accession 243 of the MOVC carries the N(1) locus, which is dominant to the presence of seed coat fuzz. No variation from expected ratios was observed in the F(2), BC(1)F(1), F(2:3), and BC(1)F(2) populations of the DP 5690 x 243 cross. The N(3) allele had no apparent effect on the expression of the N(1) locus. In a cross between accessions 243 x 143, a few plants were observed which were completely devoid of lint and fuzz fiber (fiberless). A fiberless line was developed from one of these fiberless plants. This line was designated MD 17 fiberless. In a cross between DP 5690 x MD 17 fiberless, we demonstrated that at least three loci were involved in the expression of the fiberless phenotype. The involvement of n(2) and n(3) in the expression of this fiberless phenotype was demonstrated in the F(2) progeny of the cross between 143 x MD 17 fiberless. This is the first demonstration that N(1), n(2), and n(3) interacted to produce fiberless seed. PMID- 12547924 TI - Genetic mapping of the evergrowing gene in peach [Prunus persica (L.) Batsch]. AB - In temperate locations, terminal apices on evergrowing (also called evergreen) peach trees keep growing in winter until killed by low temperatures, while the lateral buds go into dormancy. A recessive allele of a single gene (evergrowing or evg) controls this trait in peach. The amplified fragment length polymorphism (AFLP) technique and bulked segregant analysis were applied to construct a local genetic linkage map for the evg gene from the cross Empress op op dwarf x Evergrowing (P.I. 442380). This map, comprising nine AFLP markers and the evg locus, covers a total genetic distance of 79.3 cM. Four dominant AFLP markers (EAT/MCAC, ETT/MCCA2, EAT/MCTA, and ETT/MACC) were linked to the evg locus at distances of 1, 5.3, 6.7, and 11.7 cM, respectively. EAT/MCAC and EAT/MCTA were converted into polymorphic sequence-tagged sites. Microsatellite markers in the evg region were developed from peach bacterial artificial chromosome (BAC) clones that hybridized to the AFLP marker fragments. Using three microsatellite anchor markers (pchgms12, pchgms17, and pchgms19), the local genetic linkage map was integrated into one minor linkage group of a previously constructed peach rootstock genetic linkage map. Three AFLP markers from the rootstock genetic linkage map were found linked to the evg locus. PMID- 12547926 TI - Genetic diversity in natural populations of Piper cernuum. AB - Piper cernuum is a native plant of the Brazilian Atlantic rain forest. This work studies the distribution of allozyme diversity in P. cernuum natural populations in order to establish a strategy for sustainable management and conservation. Leaf samples were collected in two Brazilian states. High divergences among populations (F(SR) = 0.380) and low divergences among regions (F(RT) = -0.069) and among gaps of the same population (F(GT) = 0.062) were found. No association between the geographical variation and the genetic distance was detected. An excess of heterozygotes was detected in the populations (F(IS) = -0.170), suggesting selection in favor of heterozygotes. The results, and the fact that the species depends on constant gap formation for maintenance of its dynamism, suggest that the founder effect is largely responsible for the structuring of populations. For sustainable management, the maintenance of plants/reproductive branches in the gaps is of major importance. The genotypes produced in these gaps are responsible for the establishment of new gaps and are the foundation for new populations, maintaining the dynamics of allele movement. PMID- 12547927 TI - Linking deer browsing and terpene production among genetic identities in Chamaecyparis nootkatensis and Thuja plicata (Cupressaceae). AB - To investigate whether differential herbivore browsing reflects genetic variation in plant defense expression, variation in needle terpenes and damage caused by black-tailed deer (Odocoileus hemionus) was analyzed on yellow-cedar (Chamaecyparis nootkatensis) and western redcedar (Thuja plicata). In a 100-genet yellow-cedar population, three genets that were heavily browsed and had extremely low levels of monoterpenes (0-0.36% dry matter), sesquiterpenes, and diterpenes were compared to unbrowsed genets (0.85-3.83% monoterpenes in dry matter). These differences were maintained in individuals protected from browsing, suggesting genetically based variation in constitutive terpene production. In western redcedar, heavily browsed trees had significantly lower total monoterpene concentrations (1.69% dry matter) than lightly browsed trees (3.32% dry matter). One heavily browsed tree expressed no monoterpenes. No differences were found for diterpenes. In both species, the genotypes with extremely low monoterpene concentrations came from the same open-pollinated families. PMID- 12547928 TI - Microsatellite analysis of genetic diversity in wild and farmed Emus (Dromaius novaehollandiae). AB - The emu (Dromaius novaehollandiae) occupies most regions of the Australian continent and in recent times has been farmed for meat, oil, and leather. Very little is known about the genetic structure of natural or farmed populations of these birds. We report a preliminary study of genetic variation in emus undertaken by typing birds from five farms and two natural populations at five polymorphic microsatellite loci. Genetic diversity was high for all populations and there was little evidence of inbreeding, with most populations conforming to Hardy-Weinberg equilibrium for most loci. Significant heterozygote deficiencies at one locus in a number of populations were detected and may indicate the presence of null alleles. Comparisons of allele frequencies showed little evidence of genetic differentiation either among farmed populations or between farmed and natural populations. PMID- 12547929 TI - Inheritance of a novel flaccid mutant in Capsicum annuum. AB - A mutant that causes a novel flaccidity phenotype in bell pepper (Capsicum annuum L.) was generated by treating seeds with ethyl methanesulfonate (EMS). Inheritance studies indicated that the mutant was controlled by a single recessive gene. It is proposed that the gene designation representing this mutation be flc (flaccid). The mutation may be useful for investigations of the genetic basis for turgor maintenance and drought stress physiology. PMID- 12547930 TI - Popmin: a program for the numerical optimization of population sizes in marker assisted backcross programs. PMID- 12547931 TI - SHAPE: a computer program package for quantitative evaluation of biological shapes based on elliptic Fourier descriptors. PMID- 12547934 TI - Vacuolar-type proton pump in the basolateral plasma membrane energizes ion uptake in branchial mitochondria-rich cells of killifish Fundulus heteroclitus, adapted to a low ion environment. AB - We examined the involvement of mitochondria-rich (MR) cells in ion uptake through gill epithelia in freshwater-adapted killifish Fundulus heteroclitus, by morphological observation of MR cells and molecular identification of the vacuolar-type proton pump (V-ATPase). MR cell morphology was compared in fish acclimated to defined freshwaters with different NaCl concentrations: low (0.1 mmol l(-1))-, mid (1 mmol l(-1))- and high (10 mmol l(-1))-NaCl environments. MR cells, mostly located on the afferent-vascular side of the gill filaments, were larger in low- and mid-NaCl environments than in the high-NaCl environment. Electron-microscopic observation revealed that the apical membrane of well developed MR cells in low- and mid-NaCl environments was flat or slightly projecting, and equipped with microvilli to expand the surface area exposed to these environments. On the other hand, in the high-NaCl environment, the apical membrane was invaginated to form a pit, and MR cells often formed multicellular complexes with accessory cells, although the NaCl concentration was much lower than that in plasma. We cloned and sequenced a cDNA encoding the A-subunit of killifish V-ATPase. The deduced amino acid sequence showed high identity with V ATPase A-subunits from other vertebrate species. Light-microscopic immunocytochemistry, using a homologous antibody, revealed V-ATPase immunoreactivity in Na(+)/K(+)-ATPase-immunoreactive MR cells in low-NaCl freshwater, whereas the immunoreactivity was much weaker in higher NaCl environments. Furthermore, immuno-electron microscopy revealed V-ATPase to be located in the basolateral membrane of MR cells. These findings indicate that MR cells are the site responsible for active ion uptake in freshwater-adapted killifish, and that basolaterally located V-ATPase is involved in the Na(+) and/or Cl(-) absorbing mechanism of MR cells. PMID- 12547935 TI - Characterization of ryanodine receptor and Ca2+-ATPase isoforms in the thermogenic heater organ of blue marlin (Makaira nigricans). AB - A thermogenic organ is found beneath the brain of billfishes (Istiophoridae), swordfish (Xiphiidae) and the butterfly mackerel (Scombridae). The heater organ has been shown to warm the brain and eyes up to 14 degrees C above ambient water temperature. Heater cells are derived from extraocular muscle fibers and express a modified muscle phenotype with an extensive transverse-tubule (T-tubule) network and sarcoplasmic reticulum (SR) enriched in Ca(2+)-ATPase (SERCA) pumps and ryanodine receptors (RyRs). Heater cells have a high mitochondria content but have lost most of the contractile myofilaments. Thermogenesis has been hypothesized to be associated with release and reuptake of Ca(2+). In this study, Ca(2+) fluxes in heater SR vesicles derived from blue marlin (Makaira nigricans) were measured using fura-2 fluorescence. Upon the addition of MgATP, heater SR vesicles rapidly sequestered Ca(2+). Uptake of Ca(2+) was thapsigargin sensitive, and maximum loading ranged between 0.8 micro mol Ca(2+) mg(-1) protein and 1.0 micro mol Ca(2+) mg(-1) protein. Upon the addition of 10 mmol l(-1) caffeine or 350 micro mol l(-1) ryanodine, heater SR vesicles released only a small fraction of the loaded Ca(2+). However, ryanodine could elicit a much larger Ca(2+) release event when the activity of the SERCA pumps was reduced. RNase protection assays revealed that heater tissue expresses an RyR isoform that is also expressed in fish slow-twitch skeletal muscle but is distinct from the RyR expressed in fish fast-twitch skeletal muscle. The heater and slow-twitch muscle RyR isoform has unique physiological properties. In the presence of adenine nucleotides, this RyR remains open even though cytoplasmic Ca(2+) is elevated, a condition that normally closes RyRs. The fast Ca(2+) sequestration by the heater SR, coupled with a physiologically unique RyR, is hypothesized to promote Ca(2+) cycling, ATP turnover and heat generation. A branch of the oculomotor nerve innervates heater organs, and, in this paper, we demonstrate that heater cells contain large 'endplate-like' clusters of acetylcholine receptors that appear to provide a mechanism for nervous control of thermogenesis. PMID- 12547936 TI - Function of pectoral fins in rainbow trout: behavioral repertoire and hydrodynamic forces. AB - Salmonid fishes (trout, salmon and relatives) have served as a model system for study of the mechanics of aquatic animal locomotion, yet little is known about the function of non-axial propulsors in this major taxonomic group. In this study we examine the behavioral and hydromechanical repertoire of the paired pectoral fins of rainbow trout Oncorhynchus mykiss, performing both steady rectilinear swimming and unsteady maneuvering locomotion. A combination of kinematic analysis and quantitative flow visualization (using digital particle image velocimetry) enables identification of the propulsive roles played by pectoral fin motions. During constant-speed swimming (0.5 and 1.0 body length s(-1)), the pectoral fins remain adducted against the body. These fins are actively recruited, however, for a variety of maneuvering behaviors, including station holding in still water (hovering), low-speed (i.e. non-fast-start) turning, and rapid deceleration of the body during braking. Despite having a shallow pectoral-fin base orientation (the plesiomorphic teleost condition), trout are capable of rotating the fin base over 30 degrees during maneuvering, which affords the fin an impressive degree of kinematic versatility. When hovering, the pectoral fins are depressed beneath the body and twisted along their long axes to allow anteroposterior sculling. During turning and braking, the fins undergo spanwise rotation in the opposite direction and exhibit mediolateral and dorsoventral excursions. Water velocity fields and calculated momentum flows in the wake of the pectoral fins reveal that positive thrust is not generated during maneuvering, except during the retraction half stroke of hovering. Relatively large laterally directed fluid force (mean 2.7 mN) is developed during turning, whose reaction powers yawing rotation of the body (4 41 degrees s(-1)). During deceleration, the wake-force line of action falls below the center of mass of the body, and this result supports a long-standing mechanical model of braking by fishes with ventrally positioned paired fins. Despite its traditional categorization as a propulsor of limited functional importance, the salmoniform pectoral fin exhibits a diverse locomotor repertoire comparable to that of higher teleostean fishes. PMID- 12547937 TI - Involvement of G(q/11) in signal transduction in the mammalian vomeronasal organ. AB - Social behaviors of most mammals are profoundly affected by pheromones. Pheromones are detected by G-protein coupled receptors in the vomeronasal organ (VNO). To investigate the role of G alpha(q/11) in vomeronasal signal transduction pathways, microvillar membranes from murine VNO were prepared. Incubation of such membranes from prepubertal females with adult male urine results in an increase in production of inositol-(1,4,5)-trisphosphate (IP(3)). This stimulation is mimicked by GTP gamma S, blocked by GDP beta S and is tissue specific. Furthermore, use of bacterial toxins such as pertussis that lead to ADP ribosylation of the G-protein alpha subunits of G(o) and G(i2) do not block the increase in IP(3) levels but U-73122, a PLC inhibitor, blocks the production of IP(3). Studies with monospecific antibodies revealed the presence of three G proteins, G alpha(o), G alpha(i2) and G alpha(q/11)-related protein, in vomeronasal neurons, concentrated on their microvilli. Our observations indicate that pheromones in male urine act on vomeronasal neurons in the female VNO via a receptor-mediated, G alpha(q/11)-protein-dependent increase in IP(3) levels. PMID- 12547938 TI - Corneal power and underwater accommodation in great cormorants (Phalacrocorax carbo sinensis). AB - In great cormorants (Phalacrocorax carbo sinensis), corneal refractive powers, determined by photokeratometry, ranged between 52.1 diopters (52.1 D) and 63.2 D. Photorefractive reflexes, determined by infrared video photorefraction, indicated that in voluntary dives the cormorants accommodate within 40-80 ms of submergence and with myopic focusing relative to the photorefractor attained when prey was approximately one bill length from the plane of the eye. Underwater, the pupils were not constricted and retained diameters similar to those in air. These results support previously reported capacities of lenticular changes in amphibious birds yet do not fully correspond with earlier reports in terms of the coupling of iris constriction with accommodation, and time course. PMID- 12547939 TI - Odor localization requires visual feedback during free flight in Drosophila melanogaster. AB - Adult fruit flies follow attractive odors associated with food and oviposition sites through widely varied visual landscapes. To examine the interaction between olfactory and visual cues during search behavior, we recorded three-dimensional flight trajectories as individuals explored controlled sensory landscapes. When presented with the source of an attractive odor invisibly embedded in the floor of a 1 m arena, flies spend most of their time hovering back and forth over the source when flying within a randomly textured visual background but fail to localize the source when searching within a uniform white surround. To test whether flies are associating unique features of the visual background with the strength of odor cues, we flew them within arenas containing evenly spaced vertical stripes. Flies readily localized the odor when flying within visual landscapes lacking azimuthal landmarks provided that vertical edges were present. Flies failed to localize odor when flying within a background pattern consisting of horizontal stripes. These results suggest that, whereas flies do not require spatially unique visual patterns to localize an odor source, they do require visual feedback generated by vertical edges. Quantitative shifts in several components of flight behavior accompanied successful odor localization. Flies decrease flight altitude, turn more often and approach visually textured walls of the arena near an odor source. A simple model based on the statistics of flight behavior supports the hypothesis that a subtle influence on these behaviors is sufficient to lead a fly to its food. PMID- 12547940 TI - Cardiovascular responses to hypoxia and anaemia in the toad Bufo marinus. AB - Amphibians exhibit cardiorespiratory responses to hypoxia and, although several oxygen-sensitive chemoreceptor sites have been identified, the specific oxygen stimulus that triggers these responses remains controversial. This study investigates whether the cardiovascular response to oxygen shortage correlates with decreased oxygen partial pressure of arterial blood (Pa(O(2))) or reduced oxygen concentration ([O(2)]) in toads. Toads, equipped with blood flow probes and an arterial catheter, were exposed to graded hypoxia [fraction of oxygen in the inspired air (FI(O(2)))=0.21, 0.15, 0.10, 0.07 and 0.05] before and after reductions in arterial [O(2)] by isovolemic anaemia that reduced haematocrit by approximately 50%. Toads responded to hypoxia by increasing heart rate (fH) and pulmocutaneous blood flow (Q(pc)) and reducing the net cardiac right-to-left shunt. When arterial [O(2)] was reduced by anaemia, the toads exhibited a similar cardiovascular response to that observed in hypoxia. While arterial CO(2) partial pressure (Pa(CO(2))) decreased significantly during hypoxia, indicative of increased alveolar ventilation, anaemia did not alter Pa(CO(2))). This suggests that reductions in [O(2)] mediate cardiovascular adjustments, while ventilatory responses are caused by reduced Pa(O(2)). PMID- 12547941 TI - Evidence for glutamatergic mechanisms in the vagal sensory pathway initiating cardiorespiratory reflexes in the shorthorn sculpin Myoxocephalus scorpius. AB - Glutamate is a major neurotransmitter of chemoreceptor and baroreceptor afferent pathways in mammals and therefore plays a central role in the development of cardiorespiratory reflexes. In fish, the gills are the major sites of these receptors, and, consequently, the terminal field (sensory area) of their afferents (glossopharyngus and vagus) in the medulla must be an important site for the integration of chemoreceptor and baroreceptor signals. This investigation explored whether fish have glutamatergic mechanisms in the vagal sensory area (Xs) that could be involved in the generation of cardiorespiratory reflexes. The locations of the vagal sensory and motor (Xm) areas in the medulla were established by the orthograde and retrograde axonal transport of the neural tract tracer Fast Blue following its injection into the ganglion nodosum. Glutamate was then microinjected into identified sites within the Xs in an attempt to mimic chemoreceptor- and baroreceptor-induced reflexes commonly observed in fish. By necessity, the brain injections were performed on anaesthetised animals that were fixed by 'eye bars' in a recirculating water system. Blood pressure and heart rate were measured using an arterial cannula positioned in the afferent branchial artery of the 3rd gill arch, and ventilation was measured by impedance probes sutured onto the operculum. Unilateral injection of glutamate (40-100 nl, 10 mmol l(-1)) into the Xs caused marked cardiorespiratory changes. Injection (0.1-0.3 mm deep) in different rostrocaudal, medial-lateral positions induced a bradycardia, either increased or decreased blood pressure, ventilation frequency and amplitude and, sometimes, an initial apnea. Often these responses occurred simultaneously in various different combinations but, occasionally, they appeared singly, suggesting specific projections into the Xs for each cardiorespiratory variable and local determination of the modality of the response. Response patterns related to chemoreceptor reflex activation were predominantly located rostral of obex, whereas patterns related to baroreceptor reflex activation were more caudal, around obex. The glutamate-induced bradycardia was N-methyl-D-aspartate (NMDA) receptor dependent and atropine sensitive. Taken together, our data provide evidence that glutamate is a putative player in the central integration of chemoreceptor and baroreceptor information in fish. PMID- 12547942 TI - Do familiar landmarks reset the global path integration system of desert ants? AB - It is often suggested that animals may link landmark memories to a global coordinate system provided by path integration, thereby obtaining a map-like representation of familiar terrain. In an attempt to discover if desert ants form such associations we have performed experiments that test whether desert ants recall a long-term memory of a global path integration vector on arriving at a familiar food site. Ants from three nests were trained along L-shaped routes to a feeder. Each route was entirely within open-topped channels that obscured all natural landmarks. Conspicuous artificial landmarks were attached to the channelling that formed the latter part of the route. The homeward vectors of ants accustomed to the route were tested with the foodward route, either as in training, or with the first leg of the L shortened or extended. These ants were taken from the feeder to a test area and released, whereupon they performed a home vector. If travelling the latter part of a familiar route and arriving at a familiar food site triggers the recall of an accustomed home vector, then the home vector should be the same under both test conditions. We find instead that the home vector tended to reflect the immediately preceding outward journey. In conjunction with earlier work, these experiments led us to conclude in the case of desert ants that landmark memories do not prime the recall of long-term global path integration memories. On the other hand, landmark memories are known to be linked to local path integration vectors that guide ants along a segment of a route. Landmarks thus seem to provide procedural information telling ants what action to perform next but not the positional information that gives an ant its location relative to its nest. PMID- 12547943 TI - Feeding mechanisms in carp: crossflow filtration, palatal protrusions and flow reversals. AB - It has been hypothesized that, when engulfing food mixed with inorganic particles during benthic feeding, cyprinid fish use protrusions of tissue from the palatal organ to retain the food particles while the inorganic particles are expelled from the opercular slits. In crossflow filtration, the particle suspension is pumped parallel to the filter surface as filtrate exits through the filter pores, causing the suspension to become more concentrated as it travels downstream along the filter. We used high-speed video endoscopy to determine whether carp Cyprinus carpio use crossflow filtration and/or palatal protrusions during benthic feeding. We found that carp use crossflow filtration to concentrate small food particles in the pharyngeal cavity while expelling small dense inorganic particles through the opercular slits and via spits. Our results suggest that, during feeding on small food particles, palatal protrusions serve a localized chemosensory function rather than a mechanical particle-sorting function. However, palatal protrusions did retain large food particles while large inorganic particles were spit anteriorly from the mouth. We also investigated whether flow is continuous and unidirectional during suspension feeding in carp. As reported previously for ventilation in hedgehog skates and for certain industrial crossflow filtration applications, we observed that flow is pulsatile and bidirectional during feeding. These results have implications for hydrodynamic models of crossflow filtration in suspension-feeding fishes. PMID- 12547944 TI - Comparative analysis of nitric oxide and SALMFamide neuropeptides as general muscle relaxants in starfish. AB - Previous studies have established that the gaseous signalling molecule nitric oxide (NO) and the SALMFamide neuropeptides S1 and S2 cause cardiac stomach relaxation in the starfish Asterias rubens. Here we show that S1, S2 and the NO donor SNAP also cause relaxation of two other preparations from Asterias - tube feet and the apical muscle of the body wall. The rank order of effectiveness as muscle relaxants when tested at a concentration of 10 micro mol l(-1) was SNAP>S2>S1 for both tube feet and apical muscle whereas for cardiac stomach it was S2>S1>SNAP. Significantly, these data indicate that NO and SALMFamide neuropeptides function as general muscle relaxants in starfish but vary in their relative importance in different organ systems. The molecular mechanisms by which NO and SALMFamides cause muscle relaxation in starfish are not known, but previous pharmacological studies on the cardiac stomach using the soluble guanylyl cyclase inhibitor 1H-[1,2,4]oxadiazol[4,3-a]quinoxalin-1-one (ODQ) indicate that the cyclic nucleotide second messenger cGMP may mediate effects of NO. Consistent with this hypothesis, here we report that ODQ also causes partial inhibition of the relaxing effect of SNAP on tube foot and apical muscle preparations. To further investigate the involvement of cyclic nucleotides as mediators of the effects of NO and SALMFamides on starfish muscle, we have measured both cGMP and cAMP in cardiac stomach and in apical muscle after treatment with S1, S2 or SNAP. However, no significant changes in cyclic nucleotide content were observed compared with controls. Further experiments were performed on apical muscle tissue in the presence of the cyclic-nucleotide phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX), a drug that also causes cardiac stomach relaxation in starfish. Treatment with IBMX caused a 2-3 fold increase above basal levels for cGMP and cAMP, but co-treatment with IBMX and S1 or S2 or SNAP resulted in no significant further increase above the level observed with IBMX alone. We conclude from these data that the relaxing action of NO on starfish muscle may be mediated by both cGMP-dependent and cGMP-independent pathways. However, the mechanisms by which SALMFamides cause muscle relaxation in starfish remain unknown and, although our results do not rule out the involvement of cGMP or cAMP, other signalling pathways may now need to be investigated. PMID- 12547945 TI - NorpA and itpr mutants reveal roles for phospholipase C and inositol (1,4,5)- trisphosphate receptor in Drosophila melanogaster renal function. AB - Mutants of norpA, encoding phospholipase C beta (PLC beta), and itpr, encoding inositol (1,4,5)-trisphosphate receptor (IP(3)R), both attenuate response to diuretic peptides of Drosophila melanogaster renal (Malpighian) tubules. Intact tubules from norpA mutants severely reduced diuresis stimulated by the principal cell- and stellate cell-specific neuropeptides, CAP(2b) and Drosophila leucokinin (Drosokinin), respectively, suggesting a role for PLC beta in both these cell types. Measurement of IP(3) production in wild-type tubules and in Drosokinin receptor-transfected S2 cells stimulated with CAP(2b) and Drosokinin, respectively, confirmed that both neuropeptides elevate IP(3) levels. In itpr hypomorphs, basal IP(3) levels are lower, although CAP(2b)-stimulated IP(3) levels are not significantly reduced compared with wild type. However, CAP(2b) stimulated fluid transport is significantly reduced in itpr alleles. Rescue of the itpr(90B.0) allele with wild-type itpr restores CAP(2b)-stimulated fluid transport levels to wild type. Drosokinin-stimulated fluid transport is also reduced in homozygous and heteroallelic itpr mutants. Measurements of cytosolic calcium levels in intact tubules of wild-type and itpr mutants using targeted expression of the calcium reporter, aequorin, show that mutations in itpr attenuated both CAP(2b)- and Drosokinin-stimulated calcium responses. The reductions in calcium signals are associated with corresponding reductions in fluid transport rates. Thus, we describe a role for norpA and itpr in renal epithelia and show that both CAP(2b) and Drosokinin are PLC beta-dependent, IP(3) mobilising neuropeptides in Drosophila. IP(3)R contributes to the calcium signalling cascades initiated by these peptides in both principal and stellate cells. PMID- 12547946 TI - Requirement of Ca2+ on activation of sperm motility in euryhaline tilapia Oreochromis mossambicus. AB - Euryhaline tilapia Oreochromis mossambicus acclimates to the external spawning environment by modulating its mechanism for regulating sperm motility. Adaptation of sperm was performed by acclimating fish in various environments. In this paper, regulatory mechanisms of freshwater-acclimated tilapia were studied in detail. Tilapia sperm motility was vigorous in hypotonic conditions and decreased with increasing osmolality. Sperm motility was reduced in hypotonic conditions when extracellular Ca(2+) was chelated; however, extracellular Ca(2+) was not a major factor for motility activation since sperm were motile even when extracellular Ca(2+) levels were nominally depleted by EGTA. The Ca(2+) indicator, fluo 3, showed that intracellular [Ca(2+)] increased on motility activation independently of extracellular [Ca(2+)], accompanied by swelling of the sperm neck region called the sleeve structure. Intracellular [Ca(2+)] was not increased under hypertonic conditions, in which sperm were immotile, even on addition of extracellular Ca(2+). It is possible that Ca(2+) is stored in the neck region. Demembranated sperm were reactivated in the presence of Ca(2+), but cAMP failed to reactivate the motility. Furthermore, we detected phosphorylation and dephosphorylation of three proteins at serine and threonine residues on motility activation. It is likely that hypotonic shock causes an increase in intracellular [Ca(2+)] that activates motility activation via phosphorylation of some flagellar proteins. PMID- 12547947 TI - Ontogeny of feeding function in the gray short-tailed opossum Monodelphis domestica: empirical support for the constrained model of jaw biomechanics. AB - The constrained model of masticatory function enables specific predictions of bite force potentials in skulls of differing craniodental configurations. In this study, empirical support for the constrained model is provided using maximum voluntary bite force data along Region I and II of the jaws of gray short-tailed opossums Monodelphis domestica. Then, growth series of M. domestica are used to assess how bite force potential changes with growth by evaluating craniodental changes using longitudinal sets of dorsoventral radiographs and by assessing maximal bite force potential at the Region I-II boundary of the jaw in juveniles (aged 70-80 days) and adults. Our findings show that, while juveniles and adults alike enclose at least three molariform teeth within Region II (the area of highest bite force potential along the jaw), age-dependent elongation of the masticatory muscle resultant lever arm and narrowing of the palate relative to jaw length especially enhance the mechanical advantage of the adductor muscle resultant in adults. While maximal bite forces at the Region I-II boundary are absolutely greater in adults, these bite forces scale isometrically with body mass, which suggests that mass-specific forces exerted by jaw adductor muscles of larger (adult) opossums are disproportionately smaller than those exerted by smaller (juvenile) opossums. PMID- 12547948 TI - Pre-implantation genetic diagnosis. PMID- 12547949 TI - Management of child abuse in Hong Kong: results of a territory-wide inter hospital prospective surveillance study. AB - OBJECTIVES: To study suspected child abuse among children in hospital in terms of clinical characteristics and the outcome of multidisciplinary case conferences. DESIGN: Prospective observational study. SETTING: All public hospitals in Hong Kong with a paediatric department. METHODS: Anonymous data were prospectively collected from July 1997 to June 1999 using a standard report form for each case of suspected child abuse. The characteristics of the incidents and factors influencing the conclusion at the multidisciplinary case conference were studied. RESULTS: Data for 592 cases of suspected child abuse were evaluated. Two hundred and eighty-seven of the children were boys and 305 were girls. The mean age was 7.3 years (range, 0-16.7 years). Physical abuse, alone or in combination with other forms of maltreatment, accounted for 277 (86.6%) of the 320 substantiated cases. Either, or both, biological parents comprised 71.3% of the perpetrators. Seven (1.2%) children died. Of the 540 children about whom a multidisciplinary case conference was held, abuse was established for 281 (52.0%) children. Abuse was more likely to be established if the victim had been known to a childcare agency (odds ratio=2.2; 95% confidence interval, 1.4-3.5), the abuse was not sexual (odds ratio=2.7; 95% confidence interval, 1.4-5.0), or if the child was seen at a hospital that handled more than 100 cases of suspected abuse during the study period (odds ratio=3.6; 95% confidence interval, 2.4-5.4). CONCLUSION: Child abuse identified in the hospital setting is predominantly physical in nature and death is not uncommon. Appraisal of suspected child abuse by multidisciplinary case conference appears to be influenced by the region of Hong Kong in which the case was handled. PMID- 12547950 TI - Prospective randomised controlled trial comparing laparoscopic and open colposuspension. AB - OBJECTIVE: To compare the efficacy, safety, complications, and short-term outcome of laparoscopic and open colposuspension in women with genuine stress incontinence. DESIGN: Randomised controlled trial. SETTING: Urogynaecology unit in a public hospital, Hong Kong. SUBJECTS AND METHODS: Ninety patients with urodynamically proven genuine stress incontinence. Forty-three patients were randomly allocated to receive open colposuspension and 47 to undergo laparoscopy. All patients had reassessment within 1 year of the operation. MAIN OUTCOME MEASURES: Objective and subjective measures and complication rates. RESULTS: There was no significant difference in the duration of stress incontinence, mean preoperative pad test results, or proportion with pre-existing detrusor instability. Among patients in the laparoscopic group, the mean operating time was significantly longer (42.0 minutes versus 29.3 minutes; P<0.0001), while the mean blood loss was significantly less (124.7 mL versus 326.9 mL; P=0.001). Subjective and objective success rates within 1 year were similar for patients in the open and laparoscopic groups (86.0% versus 80.9%; P=0.58, and 86.0% versus 85.1%; P=1.00, respectively). There was no significant difference in the rate of complications, including de novo detrusor instability and an obstructive voiding pattern, enterocele, or dyspareunia. CONCLUSION: Laparoscopic colposuspension is a feasible alternative to the open approach. The operating time is longer but the short-term cure rate is comparable with that of the open approach. PMID- 12547951 TI - Managing older patients with urinary retention in the Continence Clinic. AB - OBJECTIVE: To evaluate the effectiveness of the Continence Clinic for managing retention of urine in older patients. DESIGN: Retrospective study. SETTING: Continence Clinic, Fung Yiu King Hospital, Hong Kong. SUBJECTS AND METHODS: Case notes of 58 patients seen at the Fung Yiu King Hospital Continence Clinic from October 1997 to September 2001 were reviewed. The patients had retention of urine with post-void residual volume of more than 200 mL, retention of urine requiring catheterization, or had catheters for unknown reasons. RESULTS: Urodynamic study performed for 22 (38%) patients showed that 12 had detrusor underactivity, six had detrusor hyperactivity with impaired contraction, and four had bladder outlet obstruction. Among the patients who were initially catheterized, the success rate for gradually stopping reliance on urinary catheterization was 84%. The success rate was higher among those who did not undergo urodynamic study than among those who had the study done (95% versus 67%; P=0.03). Reduction in post-void residual volume was observed at the last clinic visits (P<0.0001). Moreover, significant decreases in post-void residual volume were found both for patients who did and did not have urodynamic study. CONCLUSION: Most of the older patients with urinary retention with or without indwelling catheters were treated successfully in the Continence Clinic by appropriate medical therapy. Urodynamic study can be performed for selected patients when managing urinary retention. PMID- 12547952 TI - Role of magnetic resonance imaging for preoperative evaluation of patients with refractory epilepsy. AB - OBJECTIVE: To investigate the magnetic resonance imaging characteristics of patients with refractory epilepsy and the relationship to progression to surgery. DESIGN: Prospective observational study. SETTING: University teaching hospital, Hong Kong. PATIENTS: Patients undergoing preoperative evaluation for epilepsy surgery. MAIN OUTCOME MEASURE: Cranial magnetic resonance imaging findings, correlation with electroencephalographic results, and percentage of patients who were considered suitable candidates for surgery. RESULTS: Structural abnormalities associated with refractory epilepsy in 100 consecutive patients were mesial temporal sclerosis (30%), neocortical sclerosis (23%), vascular malformation (7%), neuronal migration disorders (7%), and tumours (5%). Normal brain scans were found for 28% of patients. Fourteen of 30 (46%) patients with medial temporal lobe lesions at magnetic resonance imaging were suitable candidates for surgery compared with 8/42 (19%) patients with extrahippocampal lesions (odds ratio=3.7; 95% confidence interval, 1.3-10.6; P<0.012). CONCLUSION: Mesial temporal sclerosis was the most common pathology in patients with refractory epilepsy. At the Prince of Wales Hospital, for patients who have undergone a basic magnetic resonance imaging protocol and surface electroencephalography, the result of cranial magnetic resonance imaging is an important determinant for whether patients will undergo surgery. PMID- 12547953 TI - Cephalometric norms for the upper airway in a healthy Hong Kong Chinese population. AB - OBJECTIVE: To obtain normative data for cephalometric measurements of the upper airway in the local Chinese population. DESIGN: Observational study. SETTING: University department and teaching hospital out-patient clinic. SUBJECTS AND METHODS: Subjects included 74 healthy patients, 29 males (age range, 18-35 years) and 45 females (age range, 16-42 years), with normal skeletal facial profile, no history of snoring, sleep apnoea, upper airway disease, tonsillectomy or adenoidectomy, obesity, or pathology in the pharynx. Twenty cephalometric airway measurements, including size of the tongue, soft palate, nasopharynx, oropharynx, hypopharynx, and relative position of the hyoid bone and valleculae were obtained. Landmarks on cephalometric radiographs were digitised and measurements were made using a specially designed computer programme. Error analysis of measurements was performed and comparison of measurements according to sex was made. RESULTS: Significant sex dimorphism was seen for the majority of measurements, with the exception of minimal depth of the airway, oropharyngeal depth of the airway, and the soft palate angle with the hard palate. CONCLUSION: A minimum sagittal dimension of the upper airway was evident despite differences in measurements between sexes. Findings from this study should be a useful reference for the assessment of sleep apnoea in the local population. PMID- 12547954 TI - Are periodontal diseases risk factors for certain systemic disorders--what matters to medical practitioners? AB - OBJECTIVE: To review recent data on the nature and pathobiology of periodontal infections and to elaborate how periodontal infections might increase susceptibility to some important systemic diseases and conditions. DATA SOURCES: Medline literature search and websites of the American Academy of Periodontology and American Medical Association. STUDY SELECTION: Literature and data on periodontal diseases and their links to systemic diseases. DATA EXTRACTION: Review of relevant information and data. DATA SYNTHESIS: Periodontal diseases, including gingivitis and periodontitis, are among the most common infections of humans. They are induced by bacteria and bacterial products of dental plaque and are characterised by inflammatory destruction of tooth-supporting connective tissues and alveolar bone. A growing body of scientific evidence has shown that severe periodontitis may enhance susceptibility to certain important systemic diseases and conditions, for example, cardiovascular disease, diabetes mellitus, adverse pregnancy outcomes, and pulmonary infections. The clinical implications of the emerging specialty of periodontal medicine for dental and medical practitioners are postulated. CONCLUSIONS: Periodontal diseases may be risk factors for cardiovascular disease, diabetes mellitus, adverse pregnancy outcomes, and pulmonary infections. Dental and medical practitioners should be aware of the clinical implications of these inter-relationships and treat affected patients in collaboration for better oral and general health. PMID- 12547955 TI - All that wheezes is not asthma. AB - Asthma is a common disease. Wheezing is not pathognomonic of asthma, however. One must be alert when appropriate asthmatic treatment does not provide adequate control. Other causes of airway obstruction must be considered, especially when stridor is heard. This report describes an elderly patient who had been managed as having asthma but had an endotracheal tumour. PMID- 12547956 TI - Pre-implantation genetic diagnosis in Hong Kong. AB - This paper presents the first two successful cases of pre-implantation genetic diagnosis in Hong Kong and discusses the indications and the advantages over prenatal diagnosis. Patients should be informed about the procedure and extensively counselled about the possibility of misdiagnosis and the need for conventional prenatal diagnosis during pregnancy. PMID- 12547957 TI - Through-the-scope stent for malignant gastric outlet obstruction. AB - This report is of the technique and results for through-the-scope stent in palliating malignant gastric outlet obstruction for 17 patients. All procedures were done using conscious sedation and fluoroscopy. Enteral Wallstents with a diameter of 20 mm or 22 mm and length 60 mm or 90 mm were used and delivered over a guidewire through an endoscope with an operating channel of at least 3.7 mm. A total of 18 stents were placed. One stent failed to be deployed. One stent migrated and required insertion of a second stent. One patient required repeat endoscopy to stop bleeding from the tumour. Through-the-scope stent relieved obstructive symptoms for 14 (82%) patients. The median dysphagia score improved from 4 to 2 after through-the-scope stent (P=0.001). The median overall survival and hospital-free survival time was 6 weeks (interquartile range, 3-9 weeks) and 4 weeks (interquartile range, 1-7 weeks), respectively. To conclude, through-the scope stent was safe and feasible, offering an alternative minimal invasive method to palliate obstructive symptoms for patients with inoperable tumours causing gastric outlet obstruction. PMID- 12547958 TI - Carotid stenting for irradiation-associated carotid stenosis 3 years after previous carotid endarterectomy. AB - Extracranial carotid stenosis is a known complication of external irradiation to the head and neck region. We report on a patient with previous carotid endarterectomy for irradiation-associated carotid stenosis. This patient developed symptomatic carotid stenosis over the ipsilateral common carotid artery proximal to the previous endarterectomy site 3 years later, and was successfully treated with carotid angioplasty and stenting. This case illustrates the importance of Duplex scan surveillance after carotid endarterectomy for patients with irradiation-associated carotid stenosis. The complimentary role of carotid endarterectomy and carotid angioplasty for managing such a patient is highlighted. PMID- 12547959 TI - Spontaneous spinal epidural haematoma--an unusual cause of spinal cord compression. AB - Spinal epidural haematoma is uncommon. Spontaneous spinal epidural haematoma of unknown aetiology is a rarity. This report is of two patients with spontaneous spinal epidural haematoma with different outcomes and discussion of the clinical picture and possible aetiology of this rare condition. PMID- 12547960 TI - Tuberculous meningitis with spinal tuberculous arachnoiditis. AB - This report is of a 36-year-old woman who initially presented with confusion and fever. Subsequent investigations showed tuberculous meningitis with acute hydrocephalus. Ventriculoperitoneal shunt was performed and anti-tuberculosis therapy was given. The patient was later noticed to have weakness of both lower limbs and urinary retention. Magnetic resonance imaging of the thoracic spine showed radiological features of tuberculous arachnoiditis with cord compression. Decompressive laminectomy was performed and high-dose systemic corticosteroid was given. A high level of awareness is required when diagnosing tuberculous arachnoiditis and the importance of high-dose corticosteroid in the treatment regimen is emphasised. PMID- 12547962 TI - Medecins Sans Frontieres experience in the provision of health care in complex settings. PMID- 12547961 TI - Moyamoya syndrome in a child with Down syndrome. AB - Moyamoya syndrome has been reported in association with Down syndrome. In paediatric patients, the usual presentation is that of ischaemic stroke. We report a 9-year-old boy with Down syndrome and moyamoya syndrome who presented with acute-onset left hemiparesis. This is the first such reported case in Hong Kong. There is growing evidence that the chromosomal abnormalities in patients with Down syndrome may contribute to a vulnerability for the development of moyamoya syndrome. A high index of suspicion is necessary to make the correct diagnosis. Medical and surgical management strategies for this disease are discussed. Surgical intervention should proceed without delay, if indicated, to prevent further neurological deterioration. A multidisciplinary approach is recommended for the rehabilitation of these patients. PMID- 12547963 TI - Personal risk management. PMID- 12547964 TI - Tracheal bronchus. PMID- 12547965 TI - Work practice in Australia. PMID- 12547966 TI - An historical overview of Australia's largest and oldest provider of vocational rehabilitation--CRS Australia. AB - CRS Australia, the largest and oldest vocational rehabilitation provider in Australia, is examined over its 60-year history. The Australian government located CRS initially in the Department of Social Security, and subsequently in the Department of Community Services however, its establishment required a complexity of policy and range of skills that were not found in one arm of government alone and required cooperation from many government departments. A disagreement between the government and the medical profession about the purpose and control of rehabilitation features in the early history of CRS Australia. Factors such as greater involvement of people with disabilities and government inquiries led to a move from a centre-based to a community-based approach to vocational rehabilitation in the 1970s. CRS Australia is a large employer of health and human service professionals and has played a significant part in the development of these professions in Australia. In recent years, CRS Australia has provided greater business accountability to government and the broader Australian community. CRS Australia's role as an expert vocational assessment agency and vocational rehabilitation provider is expected to continue as welfare reform progresses in Australia. PMID- 12547967 TI - A survey of occupational therapy in Australian work practice. AB - While the prevention and management of work-related injuries are major areas of practice for Australian occupational therapists, there has been little research on occupational therapy's contribution in these areas. This study examined the characteristics of current Australian occupational therapy practice in the work arena. A postal questionnaire was sent to 355 occupational therapists employed in work practice from six of the eight state or territory Occupational Therapy Associations. The questionnaire enquired about demographic details, service provision in the prevention, assessment and rehabilitation areas and further education needs in these areas. The responses of 125 (35%) therapists were analysed and show the services commonly offered in prevention, assessment and rehabilitation. The most commonly offered services included job analyses, workplace assessment and suitable duties programs. The results also indicate high levels of service provision in the workplace. In presenting the findings of the survey, current trends in international and Australian occupational therapy work practice are discussed along with suggestions for directions in future education, practice and research. PMID- 12547968 TI - Expert review of an approach to functional capacity evaluation. AB - Functional capacity evaluation (FCE) is a widely used tool in work rehabilitation, despite the limited examination of the soundness of its measurement properties. This paper outlines the development of a new approach to FCE, the GAPP FCE, and reports on the findings of an expert review of aspects of its content validity and technical adequacy and how it meets established test criteria. Five expert occupational therapists reviewed the materials of the GAPP FCE then completed a questionnaire related to the content validity, technical adequacy and safety, reliability, validity, practicality and utility of the GAPP FCE. The experts gave support to most aspects of these criteria. The main issue identified by the review was related to interpretation and extrapolation of the FCE results for return to work. This and other issues are discussed in relation to recent developments in FCE and plans for future development of the GAPP FCE. PMID- 12547969 TI - Test-retest reliability on nine tasks of the Physical Work Performance Evaluation. AB - The Physical Work Performance Evaluation (PWPE) is one of many functional capacity evaluations (FCEs) currently available to assist with determining injured workers' physical potential to return to work. Previous research has explored interrater reliability, construct and predictive validity of the PWPE. This research examined test-retest reliability on a sample of 24 clients with stable physical injuries who were participating in vocational rehabilitation. Nine of the 21 main tasks of the PWPE were evaluated: lifting floor to waist, bilateral carrying, pushing, sitting, standing, kneeling, stair climbing, repetitive squatting and walking. Kappa scores ranged from 0.19 (error) to 0.77 and percent agreement from 66.7% to 87.5%. The results for kneeling, lifting floor to waist, bilateral carrying and pushing tasks suggest substantial test retest reliability with moderate reliability also suggested for the standing and repetitive squatting tasks. Self-limiting behavior and alterations in pain scores, position adjustments and movement deviations are seen to be the main contributors to affect scoring between the first and second tests. PMID- 12547970 TI - Cognitive and behavioural assessment of people with traumatic brain injury in the work place: occupational therapists' perceptions. AB - Cognitive and behavioural impairments, in the absence of severe physical disability, are commonly related to poor return to work outcomes for people with traumatic brain injury (TBI). Along with other health professionals, occupational therapists make judgements about cognitive and behavioural dimensions of work capacity of clients with TBI during the return to work process. Unlike many physical functional capacity evaluations, there is no standard method that therapists use to assess the ability of people with TBI to perform cognitive operations required for work. Little is known about what information occupational therapists use in their assessment of cognitive and behavioural aspects of client performance within the work place. This study employed qualitative research methods to determine what information is utilised by 20 therapists who assess the work capacity of people with TBI in the workplace. Results indicated that the process of making judgements about cognitive and behavioural competence within the work place is a multifaceted process. Therapists triangulate client information from multiple sources and types of data to produce an accurate view of client work capacity. Central to this process is the relationship between the client, the job and the work environment. PMID- 12547971 TI - Cross-disciplinary competency standards for work-related assessments: communicating the requirements for effective professional practice. AB - The purpose of this article is to introduce the cross-disciplinary competency standards for work-related assessments, why they are needed and how they have been developed in New South Wales (NSW). Cross-disciplinary competency standards communicate the benchmarks for effective performance of work-related assessments. They outline what is expected of rehabilitation professionals, including the ability to apply and transfer competence across different conditions and workplace contexts. Outcomes in occupational rehabilitation are affected by the efficacy of the work-related assessments performed, which is dependent upon competent, clinical decision-making by rehabilitation professionals. However, in Australia, work-related assessment practice is not governed by universally accepted competency standards or by any competency-based training/education and assessment system. To enhance professional practice, WorkCover NSW has developed cross-disciplinary competency standards for work-related a ssessments. The competencies provide (i) quality standards for professional workplace training and development, (ii) benchmarks for assessing the competence of rehabilitation professionals, (iii) a framework for evidence-based practice, (iv) benchmarks for measuring service quality and (v) "real world" learning outcomes and assessment criteria for professional education programs. PMID- 12547972 TI - Examining the effectiveness of health and safety committees and representatives: a review. AB - The establishment of a health and safety committee in the workplace and/or the appointment of an elected health and safety representative is believed to foster employee participation and consultation in the workplace. Despite an emphasis within the Australian occupational health and safety legislation towards employee participation in relation to health and safety issues, there is little known about the effectiveness of the strategies used to involve workers in health and safety matters in the workplace. There is anecdotal evidence published in relation to the implementation of health and safety committees and their effectiveness in the Australian workplace. Few opinions however are substantiated by empirical evidence. Significantly more research has been conducted on the role of health and safety representatives in Australia. This paper presents the findings of a review of the literature and identifies the factors that are considered to impact on the effectiveness of workplace health and safety committees and representatives. Several factors are described as being fundamental to the effective performance of committees and representatives. These factors include management commitment, communication, training and information, union involvement, the infrastructure of an organisation, committee processes and the involvement of professional experts. It is concluded that the effectiveness of health and safety committees in Australia is unexplored and it is unknown if these committees achieve the purpose for which they were established. Recommendations are made for future research to be conducted to test the current anecdotal evidence and assumptions associated with the operation and effectiveness of health and safety committees. PMID- 12547973 TI - When is it work? Perceptions of six-year-old children. AB - A naturalistic study was undertaken to explore six-year-old children's perceptions of work in their school day. Twenty-four Year One children enrolled in an infants' school, located in Southern Sydney were involved. Participant observation and focus group interviewing were used to elicit descriptive information. A fishing game, drawing activity and excerpts from a videotape of their day at school were used as stimuli to capture how children categorised and attributed meaning to their own work performance. The findings revealed that these children had well-developed ideas about what is work. They employed a highly individual classification process to determine which occupations were work versus play, self-care and rest, resulting in differences of opinion among the children. This process was based on four factors: the physical and social environment, the type of task being performed, personal meaning attributed to the task and the child's perceived level of control. PMID- 12547974 TI - The role of work in the lives of retired men following stroke. AB - A study was conducted with the purpose of describing self-perceived occupational role performance and to explore the possibility that role is a concept that is understood and utilised in the planning and performance of meaningful occupations. The participants were thirteen men who had experienced a stroke and undergone rehabilitation. The naturalistic methodology employed included a single extensive interview to establish and describe self-perceived roles and occupational role performance, followed by role sorting and role rating tasks. Data from the tasks were used to verify transcript analysis data. The findings of this study as they relate to the concept of work are described. Work as perceived by the study participants was found to continue beyond retirement and the advent of disability. Work roles, work occupations and workful elements of occupational performance were identified. For some participants a sense of being productive was highly significant to their feelings of well-being and social fit. A number of strategies were employed by participants to enable them to continue to do work. It is suggested that work should be considered in rehabilitation even beyond retirement. PMID- 12547975 TI - Evaluation and treatment of neurobehavioral complications of traumatic brain injury--have we made any progress? PMID- 12547976 TI - Evaluation and treatment of postconcussive symptoms. AB - Postconcussive symptoms such as headache, dizziness, irritability, and difficulties with memory and attention are reported frequently after traumatic brain injuries (TBI) of all severities. The etiology of these symptoms in individuals with mild TBI has been a subject of some controversy with theories ranging from neural damage to malingering. Furthermore, although the term postconcussive syndrome is commonly used clinically and in the scientific literature, it is not clear that postconcussive symptoms constitute a syndrome per se. Instead, it may be the case that the various symptoms that commonly co occur after TBI are relatively independent consequences of a single neurological event. In other words, because the locations and severity of injury vary between individuals despite ostensibly similar injuries, it follows that there should be variations in symptom type and severity between individuals as well. This article reviews the sequelae and natural course of recovery from mild TBI, the evidence regarding both persistent postconcussive symptoms and the postconcussive syndrome, and outlines an approach to the assessment and treatment of individuals with these symptoms after TBI. PMID- 12547977 TI - Lack of awareness and its impact in traumatic brain injury. AB - Research suggests that up to 45% of individuals with moderate to severe TBI demonstrate reduced awareness or complete lack of awareness of their deficits. We describe dimensions and distinctions within the concept of lack of awareness - including whether an individual has knowledge of a specific deficit, the emotional response an individual manifests to a specific deficit, the ability to comprehend the impact or consequences of the deficit on day to day life, and how an individual explains or accounts for any deficits - and various methods of assessing for lack of awareness in this population. Finally, a review of the literature studying lack of awareness in TBI, its relationship to injury severity, the impact of lack of awareness on outcome, and intervention approaches is presented. PMID- 12547978 TI - Agitation, aggression, and disinhibition syndromes after traumatic brain injury. AB - Traumatic brain injury (TBI) is frequently complicated by disinhibition and aggression. These often profound changes in personality, present obstacles to rehabilitative treatments and community reentry. Syndromal presentations may involve a loss of impulse control, spontaneous aggression, and dysphoric bipolar states. Common neuropathological findings of inferior frontal lobe dysfunction support both disinhibition and kindling models of TBI-induced aggression. Assessment of these highly disruptive symptoms requires detailed historical, clinical, and neuropsychological information to formulate appropriate strategies. Management of TBI-related aggression may involve pharmacological, environmental, and psychotherapeutic strategies that incorporate caregiver training and support. PMID- 12547979 TI - Mood disorders following traumatic brain injury. PMID- 12547980 TI - Assessment and treatment of apathy syndrome following head injury. PMID- 12547981 TI - Executive dysfunction following traumatic brain injury: neural substrates and treatment strategies. AB - Executive dysfunction is among the most common and disabling aspects of cognitive impairment following traumatic brain injury (TBI), and may include deficits in reasoning, planning, concept formation, mental flexibility, aspects of attention and awareness, and purposeful behavior. These impairments are generally attributed to frontal systems dysfunction, due either to direct insult to the frontal lobes or to disruption of their connections to other brain regions. Evaluation of executive deficits typically includes neuropsychological assessment, though adjunctive interviews can be critical in detecting subtle dysexecutive symptoms that may not be apparent on standardized testing. Rehabilitation programs emphasizing cognitive-behavioral approaches to the retraining of planning and problem-solving skills can be effective in ameliorating identified executive deficits. In addition, pharmacological approaches may be useful in addressing aspects of executive dysfunction. This review summarizes the nature of executive deficits following TBI, their neuroanatomical substrates, selected assessment and treatment strategies, and recent research findings and trends. PMID- 12547982 TI - Anxiety disorders associated with traumatic brain injuries. AB - Anxiety disorders are common in the general population and may be even more common in people with traumatic brain injuries. This article presents a review of the literature on anxiety disorders as a result of traumatic brain injury, specifically post-traumatic stress disorder, generalized anxiety disorder, obsessive compulsive disorder, and panic disorder. Our current understanding suggests that the increased frequency of anxiety disorders after TBI may reflect an overlap between brain regions vulnerable to traumatic brain injury, and the neural circuitry of these disorders. Issues regarding treatment are largely anecdotal, and much remains unsettled. More research is needed, both in terms of diagnosis and treatment. PMID- 12547983 TI - Evaluation and treatment of psychosis after traumatic brain injury. AB - A review of research studies to date suggests that psychosis is a relatively rare, but serious, complication of traumatic brain injury (TBI). Psychotic syndromes occur more frequently in individuals who have had a TBI than in the general population. Onset of symptoms can be early or late. Psychosis can occur during the period of post-traumatic amnesia, in association with post-traumatic epilepsy, in association with TBI-related mood disorders, and as a chronic, schizophrenia-like syndrome. TBI can interact with genetic vulnerability to increase the risk of developing illnesses such as schizophrenia. Thorough diagnostic assessment is the foundation of rational and effective pharmacotherapy for psychosis after TBI. Atypical antipsychotic drugs have emerged as first line drugs for treatment of psychotic disorders from all causes, including TBI. Anticonvulsant, antidepressant or other drugs may also be needed in some cases. Medication approaches must be adjusted for the particular characteristics and vulnerabilities of the patient with a TBI. PMID- 12547988 TI - Japan's contribution to research on cardiovascular disease. AB - Japan's contribution to the research on cardiovascular disease in the past decade was analyzed by accessing journal articles published in 1991-2000 in the MEDLINE database. The number of articles having an affiliation with a Japanese institution was counted and summed up as a whole and also separately for each journal with statistical time trend analysis. The proportions of randomized controlled study (RCT), case - control/cohort study, and case reports in articles from Japan were also determined. Of the total articles, Japan's contribution to the research on cardiovascular disease was 8.9% and ranked 2nd in the world. The recent increase in the contribution was not significant (p=0.25). Compared with other countries, the proportions of RCT (2.3%) and case - control/cohort study (2.3%) were smaller and that of case reports was similar (5.7%). Although the overall contribution from Japan in the field of cardiovascular disease is satisfactory, high-quality clinical research is as meager as in other clinical fields. PMID- 12547989 TI - Noninvasive coronary artery imaging by multislice spiral computed tomography. AB - Although the excellent spatial resolution of multislice spiral computed tomography (MSCT) enables the coronary arteries to be visualized, its limited temporal resolution results in poor image reproducibility because of cardiac motion artifact (CMA) and hence limits its widespread clinical use. A novel retrospectively ECG-gated reconstruction method has been developed to minimize CMA. In 88 consecutive patients, the scan data were reconstructed using 2 retrospectively ECG-gated reconstruction methods. Method 1: the end of the reconstruction window (250 ms) was positioned at the peak of the P wave on ECG, which corresponded to the end of the slow filling phase during diastole immediately before atrial contraction. Method 2 (conventional method): relative retrospective gating with 50% referred to the R-R interval was performed so that the beginning of the reconstruction window (250 ms) was positioned at the halfway point between the R-R intervals of the heart cycle. The quality of the coronary artery images was evaluated according to the presence or absence of CMA. The assessment was applied to the left main coronary artery (LMCA), the left anterior descending artery (LAD, segments #6, #7 and #8), the left circumflex artery (LCx, segments #11 and #13) and the right coronary artery (RCA, segments #1, #2 and #3). The first diagonal artery (#9-1), the obtuse marginal artery (#12-1), the posterior descending artery (#4-PD), the atrioventricular node branch (#4-AV) and the first right ventricular branch (RV) were also evaluated. Of the 88 patients, 85 were eligible for image evaluation. Method 1 allowed visualization of the major coronary arteries without CMA in the majority of patients. The LCA system (segments #5-7, #11 and #13) and the proximal portion of the RCA were visualized in more than 94% of patients. Artifact-free visualization of the distal portion of the LAD (segment #8) and RCA (#4PD and #4AV), and side branches (#9-1, #12-1 and RV) was also achieved in more than 80% of patients. On the other hand, CMA occurred frequently on images obtained by Method 2. The LCx and RCA systems were the most affected by CMA, revealing only 41% artifact-free visualization of the segment #13, 39% of #1, 15% of #2 and 32% of #3. Thus, Method 1, which avoids the ventricular motion occurring during the rapid filling and atrial contraction phases, gives superior image quality over the conventional ECG-gated reconstruction method. PMID- 12547990 TI - Incidence and clinical features of asymptomatic atrial septal defect in school children diagnosed by heart disease screening. AB - The purpose of this study was to investigate the incidence and clinical features of atrial septal defect (ASD) in school children in Japan who were diagnosed by heart disease screening. From 1989 to 1998, a questionnaire, electrocardiography (ECG) and phonocardiogram were obtained from school children when they entered their first year of elementary school (n=86,142) or junior high school (n=80,632). In this program, 33 asymptomatic ASD patients were newly diagnosed (0.020%). The ECG findings showed incomplete right bundle-branch block (79%), right axis deviation (55%), and right ventricular hypertrophy (9%). An ejection systolic murmur was audible in 30 patients (94%) and mid-diastolic murmur in 10 patients (30%). Thirty patients (90%) showed fixed split of second heart sound. Using echocardiography or catheter observation, 31 patients (94%) were judged to require closure of the ASD. Although the medical care is widely available in Japan, undetected ASD patients were not rare and importantly, most of them required closure of the defect even if they were asymptomatic. PMID- 12547991 TI - Cutaneous arteriolar thioredoxin expression in patients with heart failure. AB - Cutaneous microangiopathic lesions exist in patients with heart failure, and heart failure is associated with increased oxidative stress. Thioredoxin (TRX) is stress-inducible and has a cytoprotective effect against oxidative stress. Accordingly, to investigate whether arteriolar TRX expression was increased in the skin of patients with congestive heart failure (CHF), skin biopsies were taken at the time of cardiac catheterization, and the results were compared with those of control subjects. The diagnosis of CHF was done by cardiac catheterization with reference to elevated plasma concentrations of TRX and brain natriuretic peptide (BNP). Increased TRX expression was found in the skin biopsies of 29 of the 35 patients with CHF, but in none of the 8 control subjects; the semiquantitative grade of arteriolar TRX immunoreactivity was 2.5+/ 1.0 in patients with CHF and 1.0+/-0.0 in controls, respectively (p<0.01). The severity of arteriolar TRX expression did not correlate with the New York Heart Association functional class. These results indicate that cutaneous arteriolar TRX expression in patients with CHF may reflect the excessive oxidative stress of the peripheral circulation associated with the condition. PMID- 12547992 TI - Effect of dobutamine on regional diastolic left ventricular asynchrony in patients with left ventricular hypertrophy. AB - Dobutamine improves systolic as well as diastolic function, but its effect on left ventricular (LV) asynchrony is unknown. An on-line automated segmental motion analysis (A-SMA) system was developed, based on an automatic border detection technique, to evaluate the effect of dobutamine on LV asynchrony in patients with LV hypertrophy (LVH). Low dose (5 microg x kg (-1) x min(-1)) dobutamine stress echocardiography was performed in 15 patients with LVH and in 15 healthy subjects. Short-axis LV views were obtained and divided into 4 wedge shaped segments using A-SMA. The time - area curve and its first derivative curve in each segment were displayed. Total normalized peak filling rates (nPFR) were obtained. Systolic and diastolic asynchronies were assessed from the coefficient of variation (CV) of the regional time intervals from end diastole to the peak ejection rate (T-PER), and from end systole to the peak filling rate (T-PFR), respectively. At baseline, the CV of T-PER and T-PFR in patients with LVH were greater than those in healthy subjects (CV-T-PER: 18.8+/-9.2 vs 9.6+/-4.3%, CV-T PFR: 19.5+/-7 vs 8.1+/-4.1%, both p<0.01). During dobutamine infusion, differences among groups at baseline disappeared and systolic and diastolic asynchronies improved (CV-T-PER: 7.3+/-4.8 vs 5.7+/-2.1%, CV-T-PFR: 6.8+/-3.5 vs 5.1+/-1.3%, both p>0.05). Total nPFR increased (from 3.2+/-1.0 /s to 5.6+/-1.3 /s, p<0.01) with dobutamine infusion in patients with LVH. Dobutamine improved LV diastolic asynchrony, as evaluated by A-SMA, in patients with LVH demonstrating that the lusitropic effect of dobutamine improved LV regional diastolic asynchrony, playing an important role in the improvement of global LV diastolic filling. PMID- 12547993 TI - Comparative usefulness of beat-to-beat QT dispersion and QT interval fluctuations for identifying patients with organic heart disease at risk for ventricular arrhythmias. AB - The comparative usefulness of 10 min of beat-to-beat 12-lead QT dispersion (QTd) and QT interval variability index (QTVI) analysis for identifying patients with organic heart disease (OHD) at risk for ventricular arrhythmias was assessed in 86 subjects: 54 had OHD without a history of ventricular arrhythmias, 15 had OHD with documented ventricular tachycardia, and there were 17 controls. The following parameters were analyzed among the groups: (1) the average QTd (mean QTd), (2) the difference between the maximum and minimum QTd observed over the recording time (QTd variation), (3) the maximum difference of QTd between consecutive beats (QTd maximum), (4) the QTd standard deviation (QTd variability), and (5) QTVI, calculated in lead I or II according to an established formula: log 10 [(QTv/QTm2) / (HRv/HRm2)]. All the analyzed parameters were significantly increased in the patients with and without ventricular tachycardia when compared with the controls. QTd variation, QTd maximum and QTd variability were the only variables that remained significantly increased in the group of patients with documented ventricular tachycardia, compared with those without arrhythmia. Thus, beat-to-beat fluctuations of both the QT interval and QTd may be markers of temporal electrical instability in patients with OHD. PMID- 12547994 TI - Coronary-prone behavior among Japanese men. AB - The Japanese Coronary-prone Behavior Scale (JCBS) is a questionnaire developed by the Eastern Collaborative Group Study (ECGS), a multi-center study of coronary prone behavior among Japanese men in Japan. From the JCBS, a subscale was extracted consisting of 9 items that were independently associated with the presence of coronary artery disease (CAD) in patients undergoing coronary angiography (CAG). The subscale, named Scale C, represents job-centered lifestyle, social dominance, and suppressed overt type A behavior. To further validate the association of Scale C with CAD among Japanese men, the present study examined if such an association would be observed in a newly sampled population. The JCBS was carried out by 311 men undergoing CAG. The association between Scale C score and the presence of CAD was evaluated with logistic regression analysis that included established coronary risk factors. The Scale C score was significantly higher in subjects with CAD than in those without and stepwise multivariate logistic regression showed that the Scale C score was associated with the presence of CAD independent of age, low-density lipoprotein cholesterol, diabetes mellitus, and obesity. These results indicate that the Scale C score is independently associated with the presence of CAD even among a newly sampled population of Japanese men undergoing CAG. Scale C may reflect an important feature of coronary-prone behavior among Japanese men. PMID- 12547995 TI - Monitoring the progression of the atrial electrical remodeling in patients with paroxysmal atrial fibrillation. AB - It is important to clarify how electrical remodeling develops in clinical cases of paroxysmal atrial fibrillation (PAF), because it has been suggested that this electrophysiological phenomenon promotes an increase in the frequency of PAF. In the present study, the f-f interval during PAF was analyzed from the ambulatory ECG recordings of 21 patients with PAF (total PAF duration >2/24 h with normal atrial size) to monitor the atrial electrophysiological changes. The patients were clinically followed-up for 6 months without any antiarrhythmic drugs. Before and after the follow-up period 24-h Holter monitoring was carried out and the duration of both the PAF and the f-f intervals during the PAF episode were evaluated. In selected cases, the atrial effective refractory period (ERP) was evaluated in an electrophysiologic study before and after the follow-up period. The total PAF duration was prolonged from 187+/-50 to 223+/-79 min (p=0.034) and the f-f interval was shortened from 0.14+/-0.03 to 0.12+/-0.02 ms (p=0.003). There was an inverse relationship between the changes in total PAF duration and f f interval (p=0.027). The ERP was shortened from 214+/-15 to 194+/-5 ms (n=5, p=0.025) and there was a direct correlation between the changes in ERP and f-f interval (p=0.048). In clinical cases, the prolongation of the PAF was related to the shortening of the f-f interval during the PAF episodes and to the shortening of the atrial ERP. Electrical remodeling plays a role in promoting the development of the atrial fibrillation in patients with PAF. PMID- 12547996 TI - Clinical efficacy of efonidipine hydrochloride, a T-type calcium channel inhibitor, on sympathetic activities. AB - Dihydropyridine Ca antagonists cause reflex tachycardia related to their hypotensive effects. Efonidipine hydrochloride has inhibitory effects on T-type Ca channels, even as it inhibits reflex tachycardia. In the present study, the influence of efonidipine hydrochloride on heart rate and autonomic nervous function was investigated. Using an electrocardiogram and a tonometric blood pressure measurement, autonomic nervous activity was evaluated using spectral analysis of heart rate/systolic blood pressure variability. Three protocols were used: (1) a single dose of efonidipine hydrochloride was administered orally to healthy subjects with resting heart rate values of 75 beats/min or more (high-HR group) and to healthy subjects with resting heart rate values less than 75 beats/min (low-HR group); (2) efonidipine hydrochloride was newly administered to untreated patients with essential hypertension, and autonomic nervous activity was investigated after a 4-week treatment period; and (3) patients with high heart rate values (>/=75 beats/min) who had been treated with a dihydropyridine L type Ca channel inhibitor for 1 month or more were switched to efonidipine hydrochloride and any changes in autonomic nervous activity were investigated. In all protocols, administration of efonidipine hydrochloride decreased the heart rate in patients with a high heart rate, reduced sympathetic nervous activity, and enhanced parasympathetic nervous activity. In addition, myocardial scintigraphy with (123)I-metaiodobenzylguanidine showed significant improvement in the washout rate and H/M ratio of patients who were switched from other dihydropyridine Ca antagonists to efonidipine hydrochloride. Efonidipine hydrochloride inhibits increases in heart rate and has effects on the autonomic nervous system. It may be useful for treating hypertension and angina pectoris, and may also have a cardiac protective function. PMID- 12547997 TI - Quantitative improvement in signal-averaged electrocardiography after coronary artery bypass grafting. AB - Abnormal signal-averaged electrocardiography (SAECG) reflects slow and heterogeneous myocardial conduction, predicting ventricular arrhythmia and sudden cardiac death in patients with ischemic heart disease. The purpose of this study was to investigate the quantitative effect of coronary artery bypass grafting (CABG) on SAECG, which is still controversial, and to identify the factors that are related to it. Pre- and postoperative SAECGs were recorded in 100 patients who underwent CABG. Compared parameters included filtered QRS duration (dQRS), root mean square voltage in the terminal 40 ms of the QRS complex (RMS40), and duration of the terminal low-amplitude signal less than 40 microV (LAS40). All 3 parameters in SAECG improved significantly after CABG (dQRS: 105+/-21 ms-->99+/ 18 ms, RMS40: 55+/-45 microV-->65+/-41 microV, LAS40: 29+/-19 ms-->25+/-12 ms). The improvements in SAECG were greater in patients who underwent complete revascularization and in those without prior myocardial infarction. In conclusion, CABG improved SAECG quantitatively, even in patients with normal SAECG. However, this improving effect was variable and closely related to the presence of prior myocardial infarction and the completeness of revascularization. PMID- 12547998 TI - Effect of p53 deficiency on external vascular cuff-induced neointima formation. AB - The p53 tumor suppressor gene may act as an inhibitor of vascular neointima formation in response to injury and in the present study the effects of p53 deficiency on external vascular cuff-induced neointima formation were evaluated. Vascular neointima formation was induced by an external vascular cuff; a polyethylene tube placed around a 2 mm segment of the left femoral artery ensheathed the adventitia, but avoided direct intraluminal injury. Two weeks after cuff placement, the cuff-sheathed and contralateral control arteries without cuff from wild-type (n=10) and p53 deficient (n=8) mice were harvested and analyzed by quantitative morphometry. The areas of the lumen, intima, and media were measured in 10 cross-sections from one edge to the other of the cuffed portion, and in the corresponding 2-mm segment of the contralateral control artery. The volume ratio of the intima to media (I/M) was calculated. The contralateral control arteries without a cuff did not have intima in either wild type or p53 deficient mice. In the cuff-sheathed arteries, neointima formation of p53 deficient mice with an I/M of 93% was significantly greater than that of wild type mice with an I/M of 50% (P=0.001). The absence of p53 is associated with increased neointima formation in response to cuff injury. PMID- 12547999 TI - Experimental study on the effectiveness and safety of radiofrequency catheter ablation with the cooled ablation system. AB - Experimental in vitro and in vivo studies were performed to assess the effectiveness and safety of the cooled-tip catheter and the Cooled Ablation System, which enables the creation of deeper and wider burn lesions in the myocardial tissue using radiofrequency current. This system was confirmed to consistently create large burns by cooling the catheter tip with circulating water within the catheter, even under unfavorable conditions. On the other hand, unfavorable effects, as a result of over burning, such as explosive vaporization within the tissue (the 'pop' phenomenon), tissue carbonization, coronary artery injury and lung injury were identified. 'Pop' was difficult to predict, but it is important to know how it can be avoided. No 'Pop' was seen without first observing an impedance decrease, thus it was considered safe to decrease the radiofrequency current if the impedance began to decrease. This system will be very effective for ablation of refractory arrhythmias, such as ventricular tachycardia or atrial flutter, but it is recommended that only experienced electrophysiologists use this system to avoid serious complications. PMID- 12548000 TI - Comparative effects of carvedilol and losartan alone and in combination for preventing left ventricular remodeling after acute myocardial infarction in rats. AB - It has been verified that losartan has beneficial effects on ventricular remodeling (VRM) after acute myocardial infarction (AMI), but the effects of carvedilol alone or in combination with losartan on this condition have not been defined. The present study used rats to compare the effects of carvedilol and losartan alone and in combination for preventing VRM after AMI. After ligation of the left coronary artery, 100 surviving female Sprague-Dawley rats were randomly assigned to 1 of 4 groups: (1) AMI control (n=25), (2) carvedilol (Car, 1 mg x kg(-1) x day(-1)) (n=25), (3) losartan (Los, 3 mg x kg(-1) x day (-1)) (n=25), and (4) Car (1 mg x kg (-1). day(-1)) + Los (3 mg x kg(-1) x day (-1)) (n=25). A sham-operated group (n=17) was also randomly selected. Drugs were administered by gastric gavage for 4 weeks. After hemodynamic studies, the hearts were fixed and analyzed pathologically. Exclusive of the rats that had died or had an infarct size <35% or >55%, complete data were obtained for 65 rats, comprising AMI control (n=13), Car (n=12), Los (n=13), combination (n=14), and sham (n=13) groups. There were no significant differences in the size of infarct among the 4 AMI groups (45.8 approximately 46.7%, all p>0.05). Compared with the sham group, left ventricular (LV) end-diastolic pressure (LVEDP), volume (LVV), weight (LVW) and septal thickness (STh) were all significantly increased (all p<0.001), whereas +/-dp/dt was significantly decreased (both p<0.001) in the AMI group. In comparison with the AMI group, LVEDP, LVV, LVW and STh were all significantly decreased (LVEDP: 12.7+/-2.3, 9.7+/-2.8, and 8.6+/-3.5 mmHg vs 20.6+/-2.7 mmHg, all p<0.001; LVV: 0.74+/-0.07, 0.76+/-0.07, and 0.70+/-0.09 ml vs 0.86+/-0.05 ml, all p<0.05; LVW: 668.4+/-52.0, 702.6+/-45.4, and 683.9+/-67.7 mg vs 787.3+/-76.7 mg, p<0.05 approximately 0.001; STh: 1.57+/-0.05, 1.48+/-0.07, and 1.46+/-0.07 mm vs 1.71+/-0.04 mm, all p<0.05), whereas +/-dp/dt was significantly increased (all p<0.05) in the Car, Los, and combination groups, with LVEDP decreasing more in both Los and the combination groups than in the Car group alone (p<0.05) and STh decreasing more in the combination group than in the Car group alone (p<0.05). Carvedilol and losartan alone and in combination all prevent VRM after AMI in rats, with almost equivalent effect. PMID- 12548001 TI - Clusters of ventricular fibrillation in a patient with an implantable cardioverter difibrillator treated with amiodarone. AB - A 44 year-old man with severe left ventricular dysfunction resulting from an old myocardial infarction developed clusters of ventricular fibrillation (VF). Although coronary bypass surgery was performed and heart failure was well controlled, the VF recurred during amiodarone therapy. Despite multiple deliveries of shocks by an implantable cardioverter defibrillator, the electrical storm could not be terminated. Some substrate for rapid ventricular tachyarrhythmias, refractory to class III drugs, can lead to death from arrhythmia. PMID- 12548002 TI - Hereditary protein s deficiency with a history of recurrent myocardial infarction. AB - Protein S is a vitamin K-dependent plasma protein that inhibits the process of blood coagulation via activation of protein C, another vitamin K-dependent plasma protein A 58-year-old man with hereditary protein S deficiency had repeated attacks of acute myocardial infarction at age 52 and 58 years. PMID- 12548003 TI - Cardiac sarcoidosis mimicking right ventricular dysplasia. AB - A 59-year-old woman with skin sarcoidosis was admitted to hospital for assessment of complete atrioventricular block. Cross-sectional echocardiography showed that the apical free wall of the right ventricle was thin and dyskinetic with dilation of the right ventricle. Thallium-201 myocardial imaging revealed a normal distribution. Both gallium-67 and technetium-99m pyrophosphate scintigraphy revealed no abnormal uptake in the myocardium. Right ventriculography showed chamber dilation and dyskinesis of the apical free wall, whereas left ventriculography showed normokinesis, mimicking right ventricular dysplasia. Cardiac sarcoidosis was diagnosed on examination of an endomyocardial biopsy specimen from the right ventricle. A permanent pacemaker was implanted to manage the complete atrioventricular block. After steroid treatment, electrocardiography showed first-degree atrioventricular block and echocardiography revealed an improvement in the right ventricular chamber dilation. Reports of cardiac sarcoidosis mimicking right ventricular dysplasia are extremely rare and as this case shows, right ventricular involvement may be one of its manifestations. PMID- 12548004 TI - Platypnea - orthodeoxia syndrome with atrial septal defect. AB - A 75-year old man was referred to hospital for symptomatic hypoxemia. He did not complain of dyspnea while supine, but while sitting or standing, he experienced dyspnea with severe hypoxemia. He did not have any pulmonary diseases that could cause dyspnea. Transesophageal echocardiography revealed an atrial septal aneurysm with a small atrial septal defect (ASD) and a mild left-to-right shunt through the ASD when the patient was supine. However, when he became upright, a severe right-to-left shunt occurred and the arterial oxygen saturation decreased from 96% to 80% with dyspnea. Cardiac catheterization revealed normal pulmonary artery pressure. He was therefore diagnosed as having platypnea - orthodeoxia syndrome. Magnetic resonance imaging of the chest showed a deformity of the atrium associated with elongation of the ascending aorta. The ASD was closed surgically and the dyspnea and hypoxemia that occurred while he was upright completely resolved. PMID- 12548006 TI - Therapeutic exploitation of checkpoint defects in cancer cells lacking p53 function. AB - Cytotoxic agents form the basis of most cancer therapies. These agents primarily affect rapidly proliferating cells, so their use incurs morbidity associated with damage to tissues such as bone marrow and gastrointestinal mucosa. Clinical outcome would be improved if it were possible to develop therapeutics with more specific activity against p53-deficient cancers, which account for over 50% of all cases. p53 deficiency alters the cellular response to DNA damage in that it leaves cells with attenuated DNA damage checkpoint controls and a reduced propensity for apoptotic cell death. Thus, the DNA repair capacity of these cells is reduced but survival is increased. This promotes genomic instability and contributes to the resistance of p53-deficient cells to cytotoxic agents. Disabling the residual G(2) checkpoint function of p53-deficient cells may favour cell death following DNA damage. Several potential strategies for G(2) checkpoint abrogation show promise for the specific sensitization of cancer cells. Here we detail how the G(2) DNA damage checkpoint is influenced by p53 status and how the loss of p53 function in cancer cells can be exploited to enhance the cytotoxicity of anti-cancer agents. PMID- 12548007 TI - Cell cycle phase-specific chemotherapy: computational methods for guiding treatment. AB - Computational models of cancer chemotherapy enhance the understanding of in vitro, in vivo, and clinical trial data and have the potential to contribute to the design of rational treatment regimens. In particular, mechanistic, predictive models are superior to statistical, phenomenological descriptions of data. Mechanistic models based on functional data from tumor biopsies will enable the response to treatment to be predicted for a specific patient, in contrast to statistical models in which the probability of response for a given patient may differ substantially from the population average. This review summarizes mathematical models developed to improve the design of treatment regimens using cell-cycle phase-specific chemotherapy. It starts with simple models of dose response, then moves to more complex models of scheduling cell-cycle phase specific drugs, and finally discusses mechanistic models that incorporate both genetic drug resistance and cell cycle-mediated drug resistance. This last class of models will be most useful in designing treatment regimens tailored for individual patients. PMID- 12548008 TI - Cyclotherapy: protection of normal cells and unshielding of cancer cells. AB - Avoidance of apoptosis and mitogen-independent growth are hallmarks of cancer. Mitogen-activated kinases (for example, ErbB1, Raf-1, MEK, PI-3-K, mTOR) can suppress chemotherapy-induced apoptosis in cancer cells. While kinase inhibitors restore susceptibility of cancer cells to apoptosis, they do not necessarily cause growth arrest in cancer cells harboring additional mutations in downstream signaling pathways such as inactivation of Rb and overexpression of c-myc. This article provides a conceptual basis for a novel use of inhibitors of mitogenic kinases. While arresting growth of normal cells, kinase inhibitors may not arrest cancer cells but instead can sensitize them to apoptosis. Following pretreatment with low doses of kinase inhibitors, the chemotherapy that predominantly induces apoptosis in cycling cells (cyclotherapy) will kill cancer cells while sparing normal cells. PMID- 12548009 TI - Conversion of drug-induced differentiation to apoptosis by pharmacologic cyclin dependent kinase inhibitors. AB - Recently, considerable attention has focused on the clinical development of novel anticancer agents which are intended to induce differentiation (i.e., protein kinase C activators and histone deacetylase inhibitors) or to inhibit cyclin dependent kinases (CDKs) (i.e., flavopiridol and UCN-01). Because the differentiation process requires cell cycle arrest (e.g., in G(1)), the possibility arises that CDK inhibitors might potentiate the maturation response of neoplastic cells to various differentiation-inducing agents. However, recent findings indicate that contrary to expectations, pharmacologic CDK inhibitors fail to promote differentiation, at least in human leukemia cells; instead, they antagonize the maturation process and induce dysregulation of various cell cycle and apoptotic regulatory proteins that culminate in mitochondrial injury and apoptosis. A brief summary of the events that might contribute to these phenomena in human leukemia cells follows below. A better understanding of interactions between putative differentiation-inducers and cell cycle inhibitors may provide the foundation for the future development of novel chemotherapeutic strategies in hematopoietic and possibly non-hematopoietic malignancies. PMID- 12548010 TI - Keeping p27(Kip1) in the cytoplasm: a second front in cancer's war on p27. PMID- 12548011 TI - Are p27 and p21 cytoplasmic oncoproteins? AB - By causing cytoplasmic mislocation of p27 and p21, the Akt oncogenic kinase functionally inactivates these nuclear tumor suppressor proteins. Is cytoplasmic localization of p27 and p21 simply equivalent to loss of their function or are new functions acquired in the cytoplasm? Indeed, several lines of evidence suggest that cytoplasmic p27 and p21 may be oncoproteins with antiapoptotic activities. PMID- 12548012 TI - Understanding p27(kip1) deregulation in cancer: down-regulation or mislocalization. AB - There is considerable evidence that the inactivation of the cyclin-dependent kinase inhibitor p27(kip1) is a fundamental step for the development of human malignancies. In particular, reduced expression of p27(kip1), due to increased protein degradation, correlates with poor prognosis of patients affected by various types of cancer. The purpose of this mini-review is to present an overview of the current understanding of the alteration of p27(kip1) function in human cancer and to describe the different mechanisms that contributes to it. Particular emphasis is placed on the novel finding of p27(kip1) mislocalization in tumor cells and on the biochemical pathways responsible for p27(kip1) cytosolic accumulation. Finally, we review the possible clinical implications of these observations with respect to prognosis and novel anticancer therapies. PMID- 12548013 TI - CHK2: a tumor suppressor or not? PMID- 12548014 TI - Open heart surgery of PI-3 kinase signaling. PMID- 12548015 TI - Empty pockets yield more telomere change. PMID- 12548016 TI - Chk2 meets Plk3 in damage control. PMID- 12548017 TI - Derivatives of vitamins D2 and D3 activate three MAPK pathways and upregulate pRb expression in differentiating HL60 cells. AB - Analogs of vitamin D have been synthesized which have reduced calcemic activities yet increased anti-proliferative and differentiation-inducing properties, raising expectations that they will be useful for treatment of human neoplastic diseases. In the present study we compared the abilities of three such analogs, 24a, 24b dihomo-1,25-dihydroxyvitamin D(3) (PRI-1890), 24-ene-1,25-dihydroxyvitamin D(2) (PRI-1906) and (24R)-1,24-dihydroxyvitamin D(3) (PRI-2191) to induce markers (CD14, CD11b and MSE) of differentiation, G(1) phase block, and associated molecular events in human promyeloblastic leukemia cells HL60. We found that the potencies of the analogs to induce differentiation paralleled their activation of Erk, JNK and p38 mitogen-activated protein kinase (MAPK) pathways, and the anti proliferative activity closely correlated with the extent of hypophosphorylation of retinoblastoma protein (pRb). Interestingly, low concentrations of derivatives of vitamin D, which were insufficient to induce any detectable changes in the cell cycle traverse, markedly increased the levels of total pRb, which was highly phosphorylated. These results suggest that pRb may have an unsuspected role in monocytic differentiation, perhaps to increase the sensitivity of the G(1) checkpoint, by increasing the amount of substrate for cyclin-dependent kinases. PMID- 12548018 TI - DNMT1 is a component of a multiprotein DNA replication complex. AB - DNA methylation is a major determinant of epigenetic inheritance and plays an important role in genome stability. The accurate propagation of DNA methylation patterns with cell division requires that methylation be closely coupled to DNA replication, however the precise molecular determinants of this interaction have not been defined. In the present study, we show that the predominant DNA methyltransferase species in somatic cells, DNMT1, is a component of a multiprotein DNA replication complex termed the DNA synthesome that fully supports semi-conservative DNA replication in a cell-free system. DNMT1 protein and activity were found to co-purify with the human DNA synthesome through a series of subcellular fractionation and chromatography steps, resulting in an enrichment of methyltransferase specific activity from two human cell lines. DNA methyltransferase activity co-eluted with in vitro replication activity and DNA polymerase alpha activity on sucrose density gradients suggesting that DNMT1 is a tightly bound, core component of the replication complex. The synthesome associated pool of DNA methyltransferase exhibited both maintenance and de novo methyltransferase activity and the ratio of the two was similar to that observed in whole cell lysates and for recombinant DNMT1. These data indicate that interactions within the synthesome complex do not influence the intrinsic preference of DNMT1 for hemimethylated DNA, but suggest that newly replicated DNA may be subject to low level de novo methylation. The data indicate that DNA methylation is tightly coupled to replication through physical interaction of DNMT1 and core components of the replication machinery. The definition of the molecular interactions between DNMT1 and other proteins in the replication complex in normal and neoplastic cells will provide further insight into the regulation of DNA methylation and the mechanisms underlying the alteration of DNA methylation patterns during carcinogenesis. PMID- 12548019 TI - Genotoxic stress-induced activation of Plk3 is partly mediated by Chk2. AB - Polo-like kinase 3 (Plk3, alternatively termed Prk) is involved in the regulation of DNA damage checkpoint as well as in M-phase function. Plk3 physically interacts with p53 and phosphorylates this tumor suppressor protein on serine-20, suggesting that the role of Plk3 in cell cycle progression is mediated, at least in part, through direct regulation of p53. Here we show that Plk3 is rapidly activated by reactive oxygen species in normal diploid fibroblast cells (WI-38), correlating with a subsequent increase in p53 protein level. Plk3 physically interacts with Chk2 and the interaction is enhanced upon DNA damage. In addition, Chk2 immunoprecipitated from cell lysates of Daudi (which expressed little Plk3) is capable of stimulating the kinase activity of purified recombinant Plk3 in vitro, and this stimulation is more pronounced when Plk3 is supplemented with Chk2 immunoprecipitated from Daudi after DNA damage. Furthermore, ectopic expression Chk2 activates cellular Plk3. Together, our studies suggest Chk2 may mediate direct activation of Plk3 in response to genotoxic stresses. PMID- 12548020 TI - Farnesylated RhoB prevents cell cycle arrest and actin cytoskeleton disruption caused by the geranylgeranyltransferase I inhibitor GGTI-298. AB - Here we demonstrate that the geranylgeranyltransferase-I inhibitor GGTI-298 inhibits the RhoB pathway and disrupts stress fiber and focal adhesion formation in NIH-3T3 cells. Farnesylated (V14)RhoB-CAIM (resistant to GGTI-298), but not geranylgeranylated (V14)RhoB (-CLLL), prevented inhibition of actin stress fiber and focal adhesion formation, underlining the critical role of RhoB. In contrast, farnesylated, (V14)RhoA (-CVLS) was unable to prevent effects of GGTI 298 on cytoskeleton organization. Furthermore, the ability of GGTI-298 to induce p21(WAF) and to block cells in the G(0)/G(1) phase of the cell cycle was also prevented by farnesylated (V14)RhoB but not by farnesylated (V14)RhoA. Moreover, treatment with GGTI-298 of cells expressing farnesylated RhoB results in accumulation of these cells in the G(2)/M phase. Therefore, the RhoB pathway is a critical target of GGTI-298. PMID- 12548022 TI - Response of alveolar cells to mechanical stress. AB - The purpose of this review is to highlight areas in alveolar cell biology in which our understanding of the effects of mechanical stress have been advanced in the last year, focusing on intracellular signal transduction pathways, the surfactant system, and cell injury and repair. Mechano-transduction pathways are only now beginning to be elucidated in alveolar cells. The importance of the mitogen-activated protein kinase, G protein, and growth factor systems is emphasized. The research conducted in the last year has also stressed the importance of alveolar cell cross-talk, with surfactant exocytosis being facilitated through parathyroid hormone-related peptide and leptin and calcium in interstitial fibroblasts and endothelial cells, respectively. Finally, the importance of deformation-induced plasma membrane breaks is emphasized. Alveolar cells were found to exocytose intracellular lipid vesicles to the plasma membrane not only to prevent cell breaks but also to reseal cell breaks. This dynamic process was a stronger determinant of cell breaks than the prestress properties of the cytoskeleton. All of these exciting findings provide further potential treatment targets for ventilator-induced lung injury. PMID- 12548023 TI - Heat shock proteins and ventilator-induced lung injury. AB - In this review, we discuss the heat shock response, a specific example of gene expression that has been studied over the past 25 years, and its relevance to acute lung injury and other critical conditions. The heat shock response has been observed in virtually all organisms and involves the rapid induction of a set of highly conserved genes that encode heat shock proteins (HSPs). The HSP70 family represents the most prominent eukaryotic group of HSPs. It has been suggested that members of the HSP70 family act in the protection of cellular damage by binding to denatured or abnormal proteins after heat shock, thereby preventing protein aggregation. The capacity of HSPs to subserve cytoprotection has produced considerable interest from the perspective of elucidating the pathophysiology of organ damage and dysfunction. Several studies support the hypothesis that HSPs are cytoprotective In addition, recent investigations have demonstrated that HSP70 is released into the systemic circulation and is involved in the activation of innate immunity. PMID- 12548024 TI - Why protect the right ventricle in patients with acute respiratory distress syndrome? AB - Even a slight increase in pulmonary vascular resistance can overload a normal right ventricle, which ejects blood through a low-pressure circuit. In a clinical setting, a persistent increase in pulmonary vascular resistance produces acute cor pulmonale. From an echocardiographic point of view, may be defined as the combination of a paradoxical septal motion, reflecting systolic overload, with right ventricular enlargement, reflecting diastolic overload. In patients with acute respiratory distress syndrome, this complication reflects the severity of the pulmonary disease involving the microvasculature but may also be caused or exacerbated by an aggressive ventilatory strategy. In the past, conventional respiratory support used in acute respiratory distress syndrome to obtain normocapnia was associated with a poor prognosis and a high frequency of acute cor pulmonale, suggesting some relation between the two findings. This prognosis has greatly improved with protective ventilation. At the same time, the incidence of acute cor pulmonale has diminished in acute respiratory distress syndrome, and the prognosis of this specific complication has also improved, suggesting that the right ventricle may develop some adaptation against persistent overload. Past lessons, however, have taught us that this potential may be limited and lead us to recommend right ventricular protection during mechanical ventilation. PMID- 12548025 TI - Where are we with recruitment maneuvers in patients with acute lung injury and acute respiratory distress syndrome? AB - Reduction of tidal volume to limit plateau pressure currently is recommended for the ventilatory management of acute respiratory distress syndrome. However, sufficient evidence now exists to support the fact that excessive reduction in tidal volume may result in harmful alveolar derecruitment depending on the level at which positive end-expiratory pressure is set. The use of recruitment maneuvers has been proposed as an adjunctive lung-protective strategy to reverse low tidal volume-related derecruitment. Many questions remain regarding the basic physiologic principles of recruitment, and, therefore, the optimal way to perform recruitment maneuvers remains unknown. Moreover, apart from physiologic studies suggesting a potential benefit of recruitment maneuver in terms of recruitment and gas exchange, no data are yet available that demonstrate the ability of such a maneuver to improve outcome. In this article, we discuss the physiologic rules governing recruitment and derecruitment and review articles that provide new insights in the field of recruitment maneuver. PMID- 12548026 TI - High-frequency oscillatory ventilation in adults with acute respiratory distress syndrome. AB - The last decade has seen increased appreciation of ventilator-induced lung injury. The understanding that the process of mechanical ventilation can itself damage lungs has spurned the search for ventilation strategies that are more lung protective. High-frequency oscillatory ventilation is a mode of high-frequency ventilation that may accomplish all of the current goals of lung protection. Historically, much of the data evaluating high-frequency oscillatory ventilation came from neonatal and pediatric populations. In the past year, a number of provocative and exciting studies have been published that contribute significantly to our understanding of high-frequency oscillatory ventilation, its role in preventing and reducing ventilator-induced lung injury, and its use in the support of adult patients with lung injury. In this article, we discuss the current understanding of high-frequency oscillatory ventilation and highlight the most recent literature addressing its application in adult patients with acute respiratory distress syndrome. PMID- 12548027 TI - Unilateral lung injury. AB - Mechanical ventilation is a supportive lifesaving therapy that can potentially cause lung injury if periodic alveolar overdistension, or cyclic collapse, and reopening occur. The use of a low tidal volume with moderate to high positive end expiratory pressure improves the survival of patients with acute lung injury and acute respiratory distress syndrome. Positioning the patient with the "good lung down" and using differential ventilation with selective positive end-expiratory pressure are the two currently accepted ventilatory strategies to be applied in patients with severe unilateral lung injury. However, both have serious limitations in clinical practice. Lung injury may be rather inhomogeneous confined to one lung or preferentially distributed toward the dependent lung areas. In unilateral lung injury, ventilatory strategies that allow recruitment of injured lung and that avoid overdistension of uninjured lung parenchyma should be applied. Experimental studies have shown that the use of selective tracheal gas insufflation and partial liquid ventilation facilitates low tidal volume with appropriate gas exchange while reducing cyclic lung stretch and shear stresses. Further studies are needed to determine future applications of these therapies in humans. PMID- 12548028 TI - Effect of inspiratory time and flow settings during assist-control ventilation. AB - The effect of the settings of inspiratory time and inspiratory flow during assist control ventilation on the response of a patient's respiratory controller has been the subject of intense research during the last few years. An increase in inspiratory flow or a decrease in tidal volume delivered by the ventilator is associated with prompt increases in respiratory frequency. The changes occur before any change in arterial blood gases can take place. These responses occur both during wakefulness and sleep and in health and disease states. Whether the responses are the result of Hering-Breuer reflex activity or they arise in flow sensitive receptors remains to be defined. Important clinical implications of the respiratory rate response to changes in ventilatory settings include effects on carbon dioxide, intrinsic positive end-expiratory pressure, and, possibly, on the instability of the respiratory rate in the transition between wakefulness and sleep. PMID- 12548029 TI - Usefulness of helium-oxygen mixtures in the treatment of mechanically ventilated patients. AB - The density of helium is markedly lower than that of air or any of its components, leading to a substantial decrease in airway resistance to flow when it is inhaled. In mechanically ventilated patients with obstructive airway disease, replacing the usual air-oxygen mixture with helium-oxygen has been shown to reduce dynamic hyperinflation and intrinsic positive end-expiratory pressure; to decrease lung inflation pressures, respiratory acidosis, and work of breathing; and to improve arterial blood gases. Aerosol delivery to distal airways is enhanced with helium-oxygen. Preliminary data also suggest that the use of helium-oxygen could be a valuable approach to decrease postextubation respiratory distress. However, interference with ventilator function and added costs are two major disadvantages of helium-oxygen. Hence, before its widespread use in mechanically ventilated patients can be recommended, studies are needed to determine whether these favorable short-term effects can influence patient outcome. PMID- 12548030 TI - New modes of mechanical ventilation: proportional assist ventilation, neurally adjusted ventilatory assist, and fractal ventilation. AB - Increased knowledge of the mechanisms that determine respiratory failure has led to the development of new technologies aimed at improving ventilatory treatment. Proportional assist ventilation and neurally adjusted ventilatory assist have been designed with the goal of improving patient-ventilator interaction by matching the ventilator support with the neural output of the respiratory centers. With proportional assist ventilation, the support is continuously readjusted in proportion to the predicted inspiratory effort. Neurally adjusted ventilatory assist is an experimental mode in which the assistance is delivered in proportion to the electrical activity of the diaphragm, assessed by means of an esophageal electrode. Biologically variable (or fractal) ventilation is a new, volume-targeted, controlled ventilation mode aimed at improving oxygenation; it incorporates the breath-to-breath variability that characterizes a natural breathing pattern. PMID- 12548031 TI - Extubation failure: magnitude of the problem, impact on outcomes, and prevention. AB - Extubation failure, defined as the need for reinstitution of ventilatory support within 24 to 72 hours of planned endotracheal tube removal, occurs in 2 to 25% of extubated patients. The pathophysiologic causes of extubation failure include an imbalance between respiratory muscle capacity and work of breathing, upper airway obstruction, excess respiratory secretions, inadequate cough, encephalopathy, and cardiac dysfunction. Compared with patients who tolerate extubation, those who require reintubation have a higher incidence of hospital mortality, increased length of ICU and hospital stay, prolonged duration of mechanical ventilation, higher hospital costs, and an increased need for tracheostomy. Given the lack of proven treatments for extubation failure, clinicians must be aware of the factors that predict extubation outcome to improve clinical decision making. Risk factors for extubation failure include being a medical, multidisciplinary, or pediatric patient; age greater than 70 years; a longer duration of mechanical ventilation; continuous intravenous sedation; and anemia. Tests designed to assess for upper airway obstruction, secretion volume, and the effectiveness of cough can help to improve prediction of extubation failure. Rapid reinstitution of ventilatory support in patients who fail extubation may improve outcome. PMID- 12548032 TI - Noninvasive mechanical ventilation in acute cardiogenic pulmonary edema. AB - Acute cardiogenic pulmonary edema is a frequent life-threatening emergency. During the last 10 years, increasing attention has focused on the use of noninvasive ventilation to treat patients with various forms of acute respiratory failure. Numerous physiologic data and clinical studies support the use of noninvasive ventilation during cardiogenic pulmonary edema. Noninvasive ventilation results in rapid improvement of clinical signs of respiratory distress and gas exchange and decreases the need for endotracheal intubation for patients in the ICU with acute hypercapnic respiratory failure related to cardiogenic pulmonary edema. However, no sustained benefit (, decreased late mortality) or benefit for less severe forms of cardiogenic pulmonary edema has been demonstrated yet. Moreover, there are still few data that support the use of a specific mode of ventilation over the others. PMID- 12548033 TI - Nuclear medicine and parathyroid surgery; a change in practice. PMID- 12548034 TI - Trends in surgical techniques. AB - SUMMARY At present surgery is the only effective treatment for primary hyperparathyroidism. For many years bilateral cervical exploration has been the preferred surgical approach and has been limited to those patients with symptoms or complications of the disease. Recent improvement in preoperative localization techniques, combined with the feasibility of intraoperative parathyroid hormone measurement, has increased the opportunity to perform minimally invasive or unilateral surgery. There are now several minimally invasive procedures, including unilateral neck exploration, video assisted and complete endoscopic techniques. This review provides an overview of the different surgical approaches currently used for the management of primary hyperparathyroidism and discusses the indications, advantages and disadvantages of each technique. PMID- 12548035 TI - Biological factors influencing parathyroid localization. AB - The major factor influencing scintigraphic detection of abnormal parathyroid glands seems to be their size. However, false-negative results have been reported in large glands while some very small adenomas have been identified. Other factors can influence 99mTc-MIBI and 99mTc-Tetrofosmin uptake and therefore the accurate detection of hyperfunctioning glands depends also on these. Increases in both perfusion and functional activity and targeting of abundant mitochondria rich oxyphil cells seem to be relevant mechanisms of uptake. A relationship has been observed between the intensity of focal uptake in the parathyroid glands and the cell cycle phases for patients with secondary hyperparathyroidism. Higher uptake grades correlated with the active growing phase, showing that scintigraphy accurately reflects the functional status of the hyperplastic parathyroid glands. Serum calcium levels may modify radiotracer kinetics by influencing the membrane potential. In addition, P-glycoprotein or multidrug resistance (MDR) associated protein expression may play an important role in the false-negative results of parathyroid scintigraphy. If the lipophilic cationic radiotracers used in parathyroid scintigraphy are transported by the same mechanism as the anticancer drugs, they will be quickly eliminated from the parathyroid glands containing P glycoprotein or MDR-related protein expression and the uptake in images will be negative. In parathyroid glands with no P-glycoprotein or MDR-related protein expression, the radiotracers remain in the cells, making it easier to detect them by scintigraphy. PMID- 12548036 TI - Parathyroid imaging: preoperative localization. AB - The role of parathyroid localization is as a preoperative localization procedure. This is becoming increasingly important with the trend towards limited surgical approaches. Although there are a variety of methods used to identify the site of parathyroid adenomas, 99mTc sestamibi is at present the agent of choice. Debate continues as to whether imaging is best performed using a subtraction technique or delayed imaging/dual phase technique. There is also discussion as to which collimator to use and whether surgery should be directed by the combined use of ultrasound and radionuclide imaging. These areas are discussed within this review. The technique of choice is a subtraction 99mTc sestamibi scan of the neck using a pinhole collimator. This should be combined with a mediastinal view using a parallel hole collimator. There is sufficient evidence to suggest that combining subtraction imaging with high resolution ultrasound will improve the sensitivity and give greater information to the surgeon. PMID- 12548037 TI - Optimization of peroperative procedures. AB - Minimally invasive radio-guided surgery (MIRS) of parathyroid adenomas has been favoured by three main factors. One is the significant improvements achieved in preoperative localizing imaging, particularly with sestamibi scintigraphy. Another is the availability of intra-operative quick parathyroid hormone measurement, and finally the increase in availability of the intra-operative gamma probes in many surgical centres especially those performing sentinel node biopsy. In contrast with the traditional wide bilateral neck exploration (BNE), MIRS requires strict inclusion criteria: 1) high probability of a solitary parathyroid adenoma, 2) a significant sestamibi uptake in the parathyroid adenoma, 3) absence of a concomitant thyroid nodular disease, 4) no family history of familial hyperparathyroidism (HPT) of multiple endocrine neoplasia, 5) no previous neck irradiation. Following these criteria about 60-70% of all primary HPT patients are suitable for a MIRS. Two main protocols for MIRS have been proposed. The single day, imaging and surgery, protocol is based on the injection of a 740 MBq dose of 99mTc sestamibi with the purpose of obtaining scintigraphic imaging and then MIRS within 3 hours from radio-tracer injection. An alternative is for imaging to be performed a few days before surgery, with a further small administered activity of 37MBq of 99mTc sestamibi injected intravenously in the operating theatre a few minutes before commencing the intervention for the purpose of MIRS only. The latter protocol allows both better planning of operating theatre scheduling and reduction of the radiation exposure to the surgical staff. The main advantages of MIRS in respect to the traditional BNE include less surgical trauma, a shorter duration of anaesthesia and surgery, a shorter hospital stay with the possibility of same-day discharge, less post surgical pain with improved cosmetic results and lower costs. Moreover, MIRS has proven to be a safe technique with a low morbidity rate and a cure rate higher than 95% in patients with primary HPT. PMID- 12548038 TI - 99mTc sestamibi scintimammography in the diagnosis of palpable breast masses. AB - The aim of this prospective study was to evaluate the diagnostic performance of 99mTc sestamibi scintimammography in a region with a low incidence of breast cancer (East, central and West Africa) and to verify the clinical usefulness of this technique in identifying benign breast diseases. Thirty-eight women (age range 22-38 years) with palpable breast masses (n =38) and inconclusive mammograms were included. Prone scintimammography was performed 10 min and 60-90 min in all patients after injection using an isotime acquisition of 10 min. A positive scan for breast cancer was defined as tracer uptake on the 10 min image and retention of activity on the 60-90 min image. A negative scan was defined as uptake on the 10 min image and complete washout of activity on the 60-90 min image. All patients subsequently underwent excisional biopsy of the breast masses and histological confirmation of the pathology. The results of scintimammography and histopathology were in agreement in all patients, revealing 36 benign lesions and two malignant lesions. Histopathologically, the lesions were fibroadenomas (27), fibrocystic disease (five), abscesses (four) and invasive intraductal carcinoma (two). It is concluded that, in regions with high incidence of benign breast diseases in young women, 99mTc sestamibi scintimammography can be used to rapidly characterize benign and malignant breast masses and thereby give priority to the management of those with a serious condition. PMID- 12548039 TI - The usefulness of 99mTc tetrofosmin scintigraphy in patients with breast cancer recurrences. AB - To verify the usefulness of 99mTc tetrofosmin scintigraphy in the follow-up of breast cancer patients, we studied 72 surgically treated breast cancer patients with suspected local recurrences (20 cases) or distant metastases (52 cases) at clinical examination and/or at conventional imaging procedures (CIPs). In all patients, a whole-body scan followed by planar and single photon emission tomography (SPET) images of selected sites were acquired 10 min after the intravenous injection of 740 MBq of 99mTc tetrofosmin, using a rectangular dual head gamma camera equipped with high-resolution parallel-hole collimators. Loco regional recurrences were diagnosed in 19 patients and distant metastases in 44 cases, while benign lesions were ascertained in nine cases. 99mTc tetrofosmin SPET showed higher sensitivity, specificity and accuracy per patient than did CIP (96.8% vs 85%, 77.7% vs 55.5% and 94.4% vs 81.1%, respectively) with statistical significance for accuracy (P <0.05). The combined use of SPET and CIP achieved 100% sensitivity and 98.6% accuracy. Planar imaging did not give additional information in respect of either SPET or CIP, showing significantly lower sensitivity and accuracy values (47.6% and 52.8%, respectively). Our data seem to suggest that 99mTc tetrofosmin SPET, but not planar, may be useful in the follow up for the detection of loco-regional and distant recurrences in patients with breast cancer. The technique can play a complementary role to conventional diagnostic imaging procedures in selected patients. PMID- 12548040 TI - Effects of extracellular Na+ and Ca2+ ions and Ca2+ channel modulators on the cell-associated activity of 99mTc-MIBI and 99mTc-tetrofosmin in tumour cells. AB - Our aim was to determine whether the Ca2+ ion or cell membrane Ca2+ and Na+/Ca2+ ion transport systems are involved in maintaining the cell-associated activity of technetium-99m-hexakis-methoxy-isobutyl-isonitrile (99mTc-MIBI) and technetium 99m-ethylene-bis[bis(2-ethoxyethyl)phosphin] (99mTc-tetrofosmin) in tumour cell lines. The cell-associated activities of 99mTc-MIBI and 99mTc-tetrofosmin were assessed in various buffers, with or without Na+ and/or with different concentrations of Ca2+, in Lewi's murine lung cell carcinoma and human glioma cell lines. Different Ca2+ channel modulators, such as verapamil, flunarizine and 3,4-dichlorobenzamil (DCB), were used to assess the effect of Ca2+ channels on the cell-associated activity of 99mTc-MIBI and 99mTc-tetrofosmin. Despite significant differences between cell lines, the cell-associated activity of 99mTc MIBI was higher in buffers without extracellular Ca2+ and Na+. The cell associated activity of 99mTc-MIBI was significantly lower in all buffers containing high concentrations of Ca2+ in both cell lines. The cell-associated activity of Tc-tetrofosmin was also significantly higher in buffers without Ca2+, and was significantly decreased in buffers with high concentrations of Ca2+. All modulators significantly increased the cell-associated activity of 99mTc-MIBI in both cell lines in all buffers. All modulators increased the cell-associated activity of 99mTc-tetrofosmin, particularly in buffers containing Ca2+. The cell associated activities of both 99mTc-MIBI and 99mTc-tetrofosmin may be dependent on verapamil-, flunarizine- and DCB-sensitive Ca2+ channels. PMID- 12548041 TI - Low-dose dobutamine stress 99mTc tetrofosmin quantitative gated SPECT performed during the acute phase of myocardial infarction predicts subsequent myocardial viability and detects stunned myocardium. AB - This study evaluated the ability of low-dose dobutamine stress 99mTc tetrofosmin (DSTF) quantitative gated single photon emission computed tomography (QGS) performed during the acute phase of myocardial infarction to predict subsequent myocardial viability and detect stunned myocardium. Twenty-four patients suffering their first acute myocardial infarction (AMI) underwent coronary angioplasty after coronary angiography (CAG) immediately following admission. Follow-up CAG and left ventriculography (LVG) were performed 10 days and 6 months later. All patients underwent DSTF QGS to measure left ventricular ejection fraction (LVEF) at rest and during dobutamine infusion (10 microg.kg(-1).min(-1)) 14 days after angioplasty. No patient suffered coronary restenosis. After 6 months, the LVEF measured by LVG improved >5% in 12 patients (group A), and did not improve in the remaining 12 patients (group B). The culprit coronary artery, the peak serum creatine phosphokinase concentration, the recanalization time, and the LVEF during the acute phase were similar in the two groups. However, the increase in the LVEF was greater in group A than in group B during dobutamine infusion (deltaLVEF) as measured by DSTF QGS (11.2+/-3.8% vs 2.9+/-4.7%, P <0.001). If a cut-off value of 6.5% for the deltaLVEF was used to predict the improvement in LVEF during the chronic phase, then the sensitivity of this test was 83.3% and its specificity was 83.3%. It is concluded that DSTF QGS during AMI can be used to predict myocardial viability and detect stunned myocardium. PMID- 12548042 TI - Prognostic value of myocardial viability determined by a 201Tl SPECT study in patients with previous myocardial infarction and mild-to-moderate myocardial dysfunction. AB - The prognostic meaning of myocardial viability is most important in patients with severe left ventricular dysfunction and ischaemic heart disease, but its prognostic significance in patients with previous myocardial infarction and mild to-moderate myocardial dysfunction is uncertain. The aim of this study was to assess the prognostic value of a 201Tl single photon emission computed tomography (SPECT) rest-redistribution study in patients with previous myocardial infarction, ischaemic heart disease and mild-to-moderate myocardial dysfunction. Myocardial viability was assessed in 55 patients (50 male; mean age 58+/-9 years) by 201Tl SPECT rest-redistribution (after 4 h) scintigraphy. All patients had previous myocardial infarction (>3 months) and angiographically documented coronary artery disease, with the mean ejection fraction of 43+/-10%. Out of 55 patients, 20 were medically treated and 35 were revascularized. The follow-up period for adverse cardiac events, including death and non-fatal myocardial infarction, was 12 months. 201Tl SPECT study was positive for myocardial viability in 36 patients (65%) and negative in 19 patients (35%). Sensitivity, specificity, positive and negative predictive values for functional improvement in the follow-up period were 85%, 75%, 92% and 60%. Out of seven (13%) cardiac events in the follow-up period (four cardiac deaths and three reinfarctions), five occurred in 20 medically treated patients and two in 35 revascularized patients (25% vs 6%, P <0.05). Absence of myocardial viability was the only variable associated with adverse cardiac events (P =0.02). Survival at 12 months, as determined by using Kaplan-Meier analysis, was 56% for medically treated and non-viable patients, 80% for revascularized and non-viable patients, 91% for medically treated and viable patients, and 100% for revascularized and viable patients (P =0.0034). These findings suggest that in patients with previous myocardial infarction and mild-to-moderate myocardial dysfunction, the absence of myocardial viability as determined by the 201Tl SPECT study was the only variable associated with adverse cardiac events. The best 12 month survival was observed in revascularized viable patients, whereas the worse prognosis was found in non viable, medically treated patients. PMID- 12548043 TI - Quantitative assessment of the total myocardial uptake ratio of 123I-BMIPP by using the Ishii-MacIntyre method is useful for predicting cardiac complications in patients with mitochondrial encephalomyopathy or myotonic dystrophy. AB - We evaluated the usefulness of the total myocardial uptake ratio (TMUR) of 15-(p [123I]iodophenyl)-3(R,S)-methyl-pentadecanoic acid (123I-BMIPP) for predicting cardiac complications in patients with mitochondrial encephalomyopathy or myotonic dystrophy. Six patients with mitochondrial encephalomyopathy, four with myotonic dystrophy, and 10 control subjects were studied. Quantitative assessment of 123I-BMIPP dynamic myocardial imaging was performed, and the TMUR of 123I BMIPP was calculated according to the Ishii-MacIntyre method. Then, the TMUR was compared in the 10 patients and 10 healthy controls, and all patients were followed for 56.1+/-22.1 months to evaluate cardiac complications. TMUR in patients (2.69+/-0.64) was significantly (P =0.01) lower than that in controls (3.28+/-0.25). Three patients in whom the TMUR value was above 3.00 had no cardiac complications. On the other hand, all patients in whom TMUR was below 3.00 had some kind of cardiac complication during the follow-up period. Two patients showed progressive conduction abnormality and underwent pacemaker implantation, one patient had sick sinus syndrome and underwent pacemaker implantation, another patient showed non-sustained ventricular tachycardia and paroxysmal atrial fibrillation, and four of seven patients, including one with a pacemaker, showed an increased cardiothoracic ratio value over 50%. In conclusion, measurement of the TMUR by the Ishii-MacIntyre method is useful for evaluating the development of cardiac complications in patients with mitochondrial encephalomyopathy or myotonic dystrophy. PMID- 12548044 TI - 99mTc-SnF2 colloid "LLK": particle size, morphology and leucocyte labelling behaviour. AB - 99mTc-SnF2 colloid (Radpharm LLK) leucocyte labelling agent is used in whole blood, exploiting phagocytosis. The objectives of this work were to optimize leucocyte labelling in leucocyte-enriched plasma, and to investigate: (i) the effect of temperature and other factors on labelling efficiency; (ii) the selectivity for different leucocyte types; (iii) the viability of the labelled cells and efflux of the radiolabel; and (iv) the physical characteristics of the colloid. Density gradient centrifugation was used to investigate the labelling efficiency, cell selectivity and efflux, Trypan blue to study the viability, and laser scattering, electron microscopy and membrane filtration to investigate particle size and morphology. Particles appeared as loose, coiled, chain-like aggregates of much smaller particles (<0.05 microm). The aggregate diameter ranged from <0.1 to >5 microm and increased with time. The distribution of radioactivity amongst the particle sizes varied widely. The labelling efficiency in leucocyte-rich plasma was enhanced at 37 degrees C compared to room temperature, and by centrifuging during labelling. The selectivity for different leucocyte types varied markedly between batches and blood samples, in some cases showing preference for mononuclear cells and in others for granulocytes. Viability was excellent and comparable with 99mTc-hexamethylpropyleneamine oxime (99mTc-HMPAO)-labelled cells. A significant fraction of radiolabel, comparable to that observed with 99mTc-HMPAO, was lost from leucocytes during incubation in vitro over 4 h. Thus, 99mTc-SnF2 is a convenient, efficient labelling agent for leucocytes, but shows variable cell selectivity which may be linked to particle size variability, and there is significant efflux of radioactivity from labelled cells. PMID- 12548045 TI - Radioiodine lobar ablation as an alternative to completion thyroidectomy in patients with differentiated thyroid cancer. AB - This study seeks to evaluate the role of radioiodine in the ablation of the remaining thyroid lobe, following a histopathological diagnosis of minimally invasive follicular carcinoma or papillary carcinoma of > or =1.5 cm size in patients undergoing hemithyroidectomy. There were 93 patients (69 females and 24 males) with an average age of 37.3+/-12.5 years (range, 16-70 years) and a mean follow-up duration of 46 months. Sixty-six of the patients had papillary cancer and remaining 27 had follicular thyroid cancer. The mean 24 h radioiodine neck uptake at the first visit was 17.2+/-7.3% (4.4-34%). In view of the large amount of thyroid tissue to be ablated, which may produce radiation induced thyroiditis, low doses of radioiodine (15-60 mCi) were administered to the patients. The patients were evaluated 6 months after radioiodine therapy with a 131I whole-body scan and 48 h radioiodine neck uptake, and a thyroglobulin assay after 4-6 weeks of levothyroxine withdrawal. The thyroid lobe was completely ablated in 53 patients (56.9%) after one dose of I and the remaining patients had partial thyroid ablation, with the mean radioiodine neck uptake being reduced to 3.1+/ 2.4%. The mean first dose of 131I was 31.8+/-11.7 mCi; the estimated mean absorbed dose was 251.3+/-149.3 Gy (range, 120-790 Gy). Around 30% patients, in each of whom a remnant thyroid lobe was ablated with a single dose of radioiodine, received < or =200 Gy. The cumulative ablation rate was 92.1% after two doses of 131I. Only seven patients needed a third dose of 131I. In our cohort, 15 patients (16.1%) complained of throat discomfort and neck pain. All of them were managed with mild analgesics except three patients who needed additional oral prednisolone for 7-10 days to overcome neck oedema. We conclude that, although completion thyroidectomy remains the standard treatment after hemithyroidectomy in cases of differentiated thyroid cancer, radioiodine ablation of an intact thyroid lobe is possible and it can be achieved with much smaller doses of radioiodine than previously believed. Lobar ablation is an attractive alternative to surgery for those who refuse to undergo completion thyroidectomy or had complications during initial surgery. However, the long-term outcome in this subset of patients remains to be determined. PMID- 12548046 TI - Scintigraphic visualization of inflammation in neurodegenerative disorders. AB - In the past few decades, our understanding of the central nervous system has evolved from one of an immune-privileged site, to one where inflammation is pathognomonic for some of the most prevalent and tragic neurodegenerative diseases. Current research indicates that diseases as diverse as multiple sclerosis, stroke and Alzheimer's disease exhibit inflammatory processes that contribute to cellular dysfunction or loss. Inflammation, whether in the brain or periphery, is almost always a secondary response to a primary pathogen. In head trauma, for example, the blow to the head is the primary event. What typically concerns the neurologist and neurosurgeon more, however, is the secondary inflammatory response that will ensue and likely cause more neuron loss than the initial injury. This paper reviews the basic neuroinflammatory mechanisms, the potential neurotoxic mediators during activation of microglia, the brain resident macrophages, and their role in neurodegeneration. Alzheimer's disease is taken as a prototype for exploring these mechanisms, as it expresses more than 40 inflammatory mediators, it is the most extensively studied disorder in terms of immune-related pathogenesis, and because of its importance as the most prevalent type of dementia. Tools for the visualization of these neuroinflammatory processes, both structural and mainly functional, are critically reviewed and discussed. PMID- 12548047 TI - Is deconvolution applicable to renography? PMID- 12548048 TI - The Shwachman Award of the North American Society for Pediatric Gastroenterology, Hepatology and Nutrition 2002: presentation. PMID- 12548049 TI - The Shwachman Award of the North American Society for Pediatric Gastroenterology, Hepatology and Nutrition 2002: acceptance. Dietary management of the malnourished child with chronic diarrhea: both nurture and nature. PMID- 12548050 TI - Gastro-esophageal reflux disease and bile acids. PMID- 12548051 TI - Bacterial-enterocyte crosstalk: cellular mechanisms in health and disease. PMID- 12548052 TI - Oribasius (fourth century) and early Byzantine perinatal nutrition. AB - BACKGROUND: The purpose of the study is the investigation of perinatal nutrition in the early Byzantine period. METHODS: The original Greek language works of the celebrated physician of the fourth century, Oribasius, were studied. RESULTS: The first Byzantine author who studied perinatal nutrition, Oribasius, provided his own concepts about the topic, focusing on the suitable choice of wet nurse and evaluation of the quality of the milk. His research proves that physicians from early Byzantine times were greatly interested in perinatal nutrition. He was aware of the knowledge of eminent ancient Greek and Roman physicians, whose extracts he quoted and opinions he generally followed. CONCLUSIONS: The study concludes that concepts of perinatal nutrition in the early Byzantine period followed those of the Hippocratic, Hellenistic, and Roman periods. The early Byzantine physicians then influenced later Byzantine and Arab physicians, and afterward, their research was introduced to European medicine. PMID- 12548053 TI - Expression, characterization, and biologic activity of recombinant human lactoferrin in rice. AB - BACKGROUND: Lactoferrin has been suggested to have many biologic activities, such as facilitating iron absorption and having antimicrobial and antiinflammatory effects. In humans, several of these activities are likely to only be facilitated by human lactoferrin because they depend on the binding of human lactoferrin to specific receptors. Rice may be a useful vehicle to introduce recombinant human lactoferrin to infant foods because it has low allergenicity and is likely to be safer than using microorganisms or transgenic animals. METHODS: Recombinant human lactoferrin was expressed in the rice cell culture system, and its biologic activity was assessed by iron-binding and -releasing properties, antimicrobial activity, and binding and uptake to Caco-2 cells. The authors also compared the stability of recombinant and native human lactoferrins against heat, low pH, and in vitro digestion. RESULTS: Biologic activity of rice-expressed recombinant human lactoferrin was similar to that of native human lactoferrin. Heat-treated proteins retained their functional activities except with severe treatment at 100 degrees C for 8 seconds, which disturbed the iron-binding capacity of recombinant human lactoferrin. Both types of proteins retained their functional activities between pH 2 and 7.4. After in vitro digestion, 50% of both proteins were detectable by enzyme linked immunosorbent assay. The remaining native and recombinant lactoferrins retained antimicrobial and Caco-2 binding and uptake activities. CONCLUSIONS: The results indicate recombinant human lactoferrin has stability similar to native human lactoferrin when exposed to thermal treatment, pH treatment, and in vitro digestion, suggesting it may be active when added to infant formula. PMID- 12548054 TI - Homocysteine prevents total parenteral nutrition (TPN)-induced cholestasis without changes in hepatic oxidative stress in the rat. AB - BACKGROUND: The role of oxidative stress in total parenteral nutrition (TPN) associated cholestasis with liver glutathione depletion was recently shown. The aims of this study were to test the appearance of cholestasis and oxidative stress during TPN, and the hypothesis that reducing oxidative stress with a precursor of glutathione (GSH), homocysteine, would restore bile flow. METHODS: Three groups of rats (weight, 179-278 g) were studied: 1) D/aa group received dextrose and amino acids (3.4 g/d); 2) D/aa/L group received the same amount of amino acids, and lipids were added on an equicaloric basis (50 kcal/d) with a lowered amount of dextrose; and 3) a control group, which received dextrose perfusion and had free access to chow. A subgroup of D/aa/L rats (n = 6) received a TPN solution containing homocysteine. After 5 days of TPN, bile was collected during 2 hours. In liver homogenates, GSH, thiobarbituric acid reactive substances (TBARS), and carbonyl content of proteins (Prot-CO) were measured to test the level of oxidative stress and hepatic lipid and protein oxidation. RESULTS: After TPN, bile flow was significantly lower in the D/aa group than in the control group. Addition of lipids further decreased bile flow. Addition of homocysteine to TPN with lipids significantly increased bile flow. Aspartate aminotransferase increased significantly in both TPN groups compared with the control group. gamma-Glutamyl transpeptidase was not different among TPN groups. An increased hepatic lipid oxidation was demonstrated by TBARS level in both TPN groups when compared with the control group. However, the liver GSH contents were not different. Protein oxidation was also significantly increased by TPN. The addition of homocysteine to TPN solution increased bile flow without liver injury or changes of lipid and protein oxidation. DISCUSSION: This study shows that TPN administered to rats induces a decrease of bile flow and an oxidative stress but that the two changes are not directly correlated. Addition of lipids further impairs bile flow but does not increase the occurrence of liver injury. Consequently, it seems more likely that TPN primarily induces a cholestatic effect that in turn induces an oxidative stress rather than inducing an oxidative stress that leads to cholestasis. However, an association of both mechanisms is not totally excluded. PMID- 12548055 TI - Management of childhood pancreatic disorders: a multidisciplinary approach. AB - INTRODUCTION: Data on therapeutic endoscopy and radiologic interventions for the management of childhood pancreatic disorders are relatively limited. This study focuses on the multidisciplinary approach to the management of pancreatitis in children. PATIENTS AND METHODS: Children with pancreatic disorders were studied from January 1992 to May 2001. Acute pancreatitis (AP) was diagnosed by clinical evaluation, serum amylase more than three times normal, and morphologic abnormalities of the pancreas on imaging. Children with recurrent abdominal pain, pancreatic calcification or ductal stones on imaging, and pancreatic ductal changes on endoscopic retrograde cholangiopancreatography (ERCP) were diagnosed with chronic pancreatitis (CP). Patients were treated by gastroenterologists, surgeons, and interventional radiologists. Pancreatic exocrine insufficiency was diagnosed in appropriate settings. RESULTS: Fifteen children--6 with AP (posttrauma, 3; gallstone disease, 1; and viral, 1), 7 with CP, and 2 with pancreatic exocrine insufficiency--were diagnosed. Local complications observed in children with AP included pseudocyst in three, and infected acute fluid collection, right-sided pleural effusion, and ascites in one patient each. Complications of AP were managed with percutaneous catheter drainage (n = 3; pseudocyst, 2; infected fluid collection, 1), additional pancreatic duct stenting (n = 2), surgical drainage (n = 1), and octreotide for pleural effusion (n = 1). Signs of CP included abdominal pain (n = 7), obstructive jaundice resulting from lower common bile duct stricture (n = 2), and bleeding from gastroduodenal artery pseudoaneurysm (n = 1). Pancreatic duct stenting relieved pain in one patient, and steel coil embolization arrested bleeding from the pseudoaneurysm. Common bile duct strictures were managed by surgical bypass (n = 2), one of which required preoperative endoscopic bile duct stenting for management of cholangitis. Two other patients with CP required no intervention. CONCLUSION: A multidisciplinary approach of radiologic and endoscopic interventions and surgery are complimentary to each other in achieving successful outcomes of complicated childhood pancreatitis. PMID- 12548056 TI - Mesenterico-left portal vein bypass in children with congenital extrahepatic portal vein thrombosis: a unique curative approach. AB - OBJECTIVES: Current management of extrahepatic portal vein thrombosis (EPVT) comprises endoscopic eradication therapy of esophageal varices and conventional shunt surgery. The authors have used the novel technique of mesenterico-left portal bypass (Rex shunt) in seven children with symptomatic EPVT, and report their results here. METHODS: Median age of the children was 12 years (range, 2-16 years). All children had portal hypertension with hypersplenism and recurrent bleeding from esophageal varices. Furthermore, one patient suffered from a severe hepatopulmonary syndrome. Preoperative evaluation included liver function tests, liver biopsy, hepatic duplex ultrasonography, and radiologic evaluation of the intrahepatic and extrahepatic vascular anatomy. The internal jugular vein was used as vein graft in all patients. RESULTS: Median follow-up period was 15 months (range, 3-28 months). Ultrasound scans revealed sufficient perfusion in all shunts (median, 35 cm/s; range, 28-60 cm/s). The intrahepatic portal perfusion in segment 4 improved from a median of 6 cm/s before surgery to 18 cm/s postoperatively. The platelet count increased within 3 months from a mean of 50,625/microL to 137,750/microL. The clinical signs of hypoxemia in the child with hepatopulmonary syndrome disappeared within 6 months. CONCLUSIONS: In accordance with the limited experience published by others, the authors' data confirmed the mesenterico-portal Rex shunt as the therapy of choice for children with EPVT. Furthermore, this report is the first to show that a hepatopulmonary syndrome can be abolished by mesenterico-portal Rex shunt. PMID- 12548057 TI - Endoscopic nodular gastritis: an endoscopic indicator of high-grade bacterial colonization and severe gastritis in children with Helicobacter pylori. AB - OBJECTIVE: To investigate the significance of endoscopic nodular gastritis associated with Helicobacter pylori infection. METHODS: This prospective study included 185 children (50.8% boys) aged 1 to 12 years (mean, 6.9 +/- 3.0 years) who underwent upper intestinal endoscopy during evaluation of chronic abdominal pain. The authors assessed the endoscopic appearance of the stomach, noting those patients with endoscopic nodular gastritis. Urease activity of gastric mucosal biopsies was measured. With histologic examination, the presence and density of H. pylori organisms, the presence of follicular gastritis, the nature of inflammation, and the gastritis activity grade and overall gastritis score were assessed. RESULTS: H. pylori infection was identified in 50 children (27%). Endoscopic nodular gastritis was significantly associated with active chronic gastritis and follicular gastritis. Nodularity in the stomach showed a high specificity (98.5%) and positive predictive value (91.7%) for the diagnosis of H. pylori infection and was observed in 22 of 50 (44%) H. pylori-positive patients and in 2 of 135 (1.5%) H. pylori-negative patients. A significant association was observed between older age and the prevalence of this finding (P< 0.001). There was a significant increase in endoscopic nodular gastritis with increased H. pylori density and a positive correlation (Pearson coefficient = 0.97) with increased gastritis score on histologic examination. Increase in gastritis score was dependent on increased H. pylori density in patients with gastric nodularity; this finding was independent of age. CONCLUSIONS: Endoscopic findings of antral nodularity in children suggest the presence of H. pylori infection and follicular gastritis and may identify cases of severe gastritis and marked bacterial colonization. PMID- 12548058 TI - Probiotic bacteria in the management of atopic disease: underscoring the importance of viability. AB - OBJECTIVES: The aim of this study was to assess the efficacy of oral supplementation of viable and heat-inactivated probiotic bacteria in the management of atopic disease and to observe their effects on the composition of the gut microbiota. METHODS: The study population included 35 infants with atopic eczema and allergy to cow's milk. At a mean age of 5.5 months, they were assigned in a randomized double-blind manner to receive either extensively hydrolyzed whey formula (placebo group) or the same formula supplemented with viable (viable LGG group) or heat-inactivated Lactobacillus GG (heat-inactivated LGG group), respectively. The changes in symptoms were assessed by the SCORAD method and the presence of some predominant bacterial genera in the feces detected with 16S rRNA specific probes. RESULTS: The treatment with heat-inactivated LGG was associated with adverse gastrointestinal symptoms and diarrhea. Consequently, the recruitment of patients was stopped after the pilot phase. Within the study population, atopic eczema and subjective symptoms were significantly alleviated in all the groups; the SCORAD scores (interquartile range) decreased from 13 (range, 4-29) to 8 (range, 0-29) units in the placebo group, from 19 (range, 4 47) to 5 (range, 0-18) units in the viable LGG group, and from 15 (range, 0-29) to 7 (range, 0-26) units in the heat-inactivated LGG group. The decrease in the SCORAD scores within the viable LGG group tended to be greater than within the placebo group. The treatments did not appear to affect the bacterial numbers within the genera enumerated. CONCLUSIONS: Supplementation of infant formulas with viable but not heat-inactivated LGG is a potential approach for the management of atopic eczema and cow's milk allergy. PMID- 12548059 TI - The insulin-like growth factor axis in children with inflammatory bowel disease. AB - BACKGROUND: The insulin-like growth factor (IGF) axis consists of two IGFs and six IGF-binding proteins (IGFBPs) that regulate proliferation and differentiation of many cell types. Malnutrition and inflammation alter the IGF axis. The authors evaluated circulating IGFs and IGFBPs in patients with inflammatory bowel disease (IBD) at the time of presentation and compared them with values obtained during remission. METHODS: Seventeen newly diagnosed pediatric IBD patients were studied on presentation and during remission. Nutritional status was assessed by body mass index (BMI) and serum protein assay. The Lloyd-Still and Green IBD clinical scoring system was used. IGF-I and IGF-II levels were measured by radioimmunoassay, and IGFBP-3 levels were measured by immunoradiometric assay. IGFBPs were quantified on ligand blots with a PhosphorImager. RESULTS: Body mass index and IBD clinical scores improved after treatment: 18.7 +/- 3.0 versus 21.3 +/- 3.0 kg/m (P = 0.023) and 74.6 +/- 16.7 versus 93.1 +/- 7.4 (P< 0.001), respectively. Protein changes were insignificant. IGFBP-3 levels increased from time of first evaluation to remission: 3,470 +/- 850 versus 4,700 +/- 473 ng/mL (P< 0.001). The ratio of IGFBP-3 to IGFBP-2 increased from first evaluation to remission: 1.7 +/- 1.9 versus 3.9 +/- 1.9 (P= 0.003). IGF-I and IGF-II levels also increased: 139 +/- 167 versus 223 +/- 118 ng/mL (P= 0.011) and 307 +/- 111 versus 386 +/- 73 ng/mL (P= 0.007), respectively. CONCLUSIONS: Circulating IGFBP 3 levels were low during active IBD and increased at remission in parallel with the IGF-I levels. The IGFBP-3 to IGFBP-2 ratio was altered in the presence of active disease in a manner that would reduce IGF-I action. This abnormality improved after treatment. PMID- 12548060 TI - Knowledge, attitudes, and practice styles of North American pediatric gastroenterologists: Helicobacter pylori infection. AB - OBJECTIVE: Most Helicobacter pylori infections are acquired during childhood. The North American Society for Pediatric Gastroenterology, Hepatology and Nutrition (NASPGHAN) recently published practice guidelines for managing pediatric H. pylori infection. Before this publication, the authors conducted a survey to assess pediatric gastroenterologists' knowledge and practices regarding H. pylori. METHODS: One hundred nine of 514 NASPGHAN members completed an Internet based questionnaire on H. pylori infection. RESULTS: Eighty-two percent of respondents performed outpatient testing for H. pylori. Of these, only 31% restricted testing to children aged >5 years. Most recommended testing for H. pylori in guideline-recommended conditions; some would not treat infected patients. Ninety-seven percent would test for H. pylori in a child with new duodenal ulcer (DU), 79% in a child with a history of DU, and 91% in a child with new gastric ulcer. However, only 86%, 60%, and 91%, respectively, would treat H. pylori infection in those conditions. A proton pump inhibitor (PPI)-based triple regimen was the first-choice therapy for 78% of respondents. Correct estimates of rates of resistance to amoxicillin, clarithromycin, metronidazole, and tetracycline were 10%, 17%, 43%, and 12%, respectively. Eighty-six percent believed there was insufficient research on H. pylori in children. CONCLUSIONS: North American pediatric gastroenterologists seem well informed about H. pylori infection in children despite the lack of published guidelines at the time of survey. Knowledge about antibiotic resistance rates was deficient. Most offered some outpatient testing for H. pylori and would test children with ulcer disease. However, some would not treat patients based on a positive result. PMID- 12548061 TI - Milk sphingomyelin accelerates enzymatic and morphological maturation of the intestine in artificially reared rats. AB - OBJECTIVES: Sphingomyelin (SPM) is the dominant phospholipid, comprising 38% of total human milk phospholipids. Although little is known about the nutritional importance of SPM during the neonatal period, SPM may affect the growth and development of tissues in the newborn infant through mechanisms regulating cell proliferation and differentiation. We evaluated the effect of sphingomyelin (SPM) in artificially reared rats as a suitable model of gut maturation in the suckling infant. METHODS: Seven-day-old Sprague-Dawley rat pups were cannulated intragastrically and reared artificially on milk containing 0.5% SPM or 0.5% phosphatidylcholine (PC) for 1 week. RESULTS: Intestinal lactase activity in the SPM group was significantly lower than that in the control or PC group. Upon histologic examination, intestinal villi were found to be occupied with vacuolated cells in the control and the PC group, whereas the vacuolated cells were restricted to the tip of villi in the SPM group. The Auerbach nerve plexus area of the ileum in the SPM group was significantly greater, possibly due to accelerated development, than that in the control group or PC group. CONCLUSIONS: The present results suggest that SPM, the dominant phospholipid in milk, plays an important role in neonatal gut maturation during the suckling period. PMID- 12548063 TI - Enteropathogenic Escherichia coli: stimulating neutrophil migration across a cultured intestinal epithelium without altering transepithelial conductance. AB - INTRODUCTION: Migration of neutrophils across the intestinal epithelium is the hallmark of inflammatory conditions of the bowel. In cultured intestinal epithelial monolayer models, neutrophils can be induced to migrate along a chemotactic gradient such as n-formyl-methionyl-leucyl-phenylalanine (fMLP). Physical passage of the neutrophils across the epithelium could disrupt the tight junctions, possibly leading to a large increase in the transepithelial conductance (G(t)). The goal of this study is to determine whether transepithelial migration of neutrophils induced by enteropathogenic (EPEC) causes changes in G(t) comparable with those seen with fMLP. METHODS: The apical side of T84 monolayers were rapidly infected with EPEC E2348/69 or exposed to 1 microM fMLP. A third group of monolayers exposed to neither EPEC nor fMLP served as control. Indium-labeled neutrophils were added to the serosal side of monolayers grown on a cell culture insert membrane (12 microm pores). G(t) was measured at fixed intervals up to 4 hours. After a 150-minute incubation, radioactivity of the neutrophils that migrated to the apical side was assayed and the number of migrating neutrophils was calculated. RESULTS: At 150 minutes, EPEC induced similar neutrophil chemotactic capability compared to fMLP (231 +/- 34.10(3) and 193 +/- 48.10(3), respectively, n = 13, P > 0.05). However, EPEC induced neutrophil migration was not associated with significant increase in G(t), 1.13 +/- 0.16 fold of baseline G(t), in distinction with fMLP groups, 13.3 +/- 0.48 fold, n = 7 (P< 0.05). G(t) changes with EPEC were seen after 4 hours of infection, but were not different in the presence or absence of neutrophil migration (1.37 +/- 0.12 fold and 1.42 +/- 0.17 fold of baseline G(t), respectively). CONCLUSIONS: The results indicate that EPEC-induced neutrophil migration can occur without significant disruption of barrier function. In addition, the chemo-attractant recruiting neutrophils during EPEC infection is unlikely to be fMLP; and, the G(t) increase seen with fMLP-driven recruitment may indicate a discretionary compromise of barrier function during neutrophil migration. PMID- 12548062 TI - A comparison of budesonide and prednisone for the treatment of active pediatric Crohn disease. AB - OBJECTIVES: Budesonide has been found effective in patients with mild and moderate Crohn disease and has been found to cause fewer side effects than prednisone. The use of oral budesonide has not been prospectively evaluated in children with Crohn disease. Therefore, the authors initiated a trial to compare remission and tolerance to budesonide and prednisone in children with mild or moderately active Crohn disease. METHODS: A prospective randomized open controlled 12-week trial was carried out comparing pH modified release budesonide, 9 mg, versus prednisone, 40 mg, in children with active mild to moderate pediatric Crohn disease. RESULTS: Thirty-three patients (20 boys and 13 girls; mean age, 14.3 years) enrolled and completed the study. The groups treated with budesonide and prednisone did not differ by age, onset of disease, location of disease, or disease activity. The remission rate at 12 weeks was 47% in the budesonide treatment group and 50% in the prednisone treatment group. Side effects occurred in 32% and 71% of patients treated with budesonide and prednisone, respectively (P< 0.05). Severity of cosmetic side effects was significantly lower in patients treated with budesonide (P< 0.01). CONCLUSIONS: Remission rates for Crohn disease with budesonide and prednisone treatment in this study were similar. Pediatric patients treated with budesonide had significantly fewer side effects than patients treated with prednisone. Budesonide should be considered an alternative to prednisone in pediatric patients with mild to moderate disease activity. PMID- 12548065 TI - Bile in the esophagus: a factor in the pathogenesis of reflux esophagitis in children. AB - OBJECTIVES: Bile reflux has been postulated to be an important factor contributing to gastroesophageal reflux disease in adults. The purpose of this study was to investigate its role in children. METHODS: Sixty-five children with symptoms of gastroesophageal reflux disease were classified on the basis of the endoscopic grade of reflux esophagitis: no esophagitis (n = 26), mild to moderate esophagitis (n = 26), and severe esophagitis (n = 13). Simultaneous 24-hour esophageal pH and bilirubin monitoring with Bilitec 2000 was performed. RESULTS: Both bile and acid reflux increased with the severity of esophagitis. The differences between all groups were significant for the percentage of total (P < 0.0005), upright (P < 0.05), and supine time (P < 0.0005) bilirubin absorbance > or = 0.14, as well as for the percentage of total and supine time pH < 4, and DeMeester score (P < 0.0005). Combined pathologic acid and bile reflux was found in 11% of children with mild esophagitis and in 70% of children with severe esophagitis, while isolated bile reflux was found in 31% and 7.5%, respectively. Combined pH and bilirubin monitoring, compared with ph-monitoring alone, increased the sensitivity from 56% to 79%, and the accuracy from 69% to 83%. CONCLUSIONS: Both bile and acid reflux increase stepwise with the severity of esophagitis. Combined acid and bile reflux is associated with severe esophagitis. Isolated acid or bile reflux is usually present in mild esophagitis. Simultaneous esophageal pH and bilirubin monitoring has a higher sensitivity, as well as predictive values and accuracy than ph monitoring alone. PMID- 12548064 TI - Efficacy and safety of oral pantoprazole 20 mg given once daily for reflux esophagitis in children. AB - OBJECTIVES: To investigate the efficacy and safety of oral pantoprazole, 20 mg (0.5 to 1.0 mg/kg/day) once daily for 28 days, in pediatric patients with reflux esophagitis. METHODS: Patients in this study (n = 15; 6 to 13 years old, 9 boys) had reflux esophagitis grade Ic or II (Vandenplas classification). The efficacy of pantoprazole to reduce esophageal acid exposure time (pH < 4), reduce the number and duration of reflux episodes, and to increase the percentage of time with gastric pH > 3 was assessed by continuous 24-hour pH monitoring. The intensity of 5 common symptoms of esophagitis was scored before and after treatment on a 4-point scale. Esophagitis was assessed at baseline and after treatment by visual inspection and by the histology of biopsies from the distal third of the esophagus. RESULTS: Before treatment, the median percentage of time with intra-esophageal pH <4 was 9.3%. After 28 days of therapy with pantoprazole, this value decreased to 2.7% (P = 0.0006). The median percentage of time with intragastric pH > 3 increased from 21% at baseline to 39% on day 28 of therapy (P = 0.005). After 28 days of treatment, all patients experienced at least partial relief from reflux symptoms. Endoscopically confirmed healing of esophagitis was seen in 47% of children (Savary-Miller classification). Histologic evidence of healing was not observed. Median serum gastrin levels were slightly elevated over baseline levels (from 74 pg/ml to 93 pg/ml). In one patient there was a transient elevation of serum GOT and GPT during treatment. CONCLUSIONS: Oral pantoprazole 20 mg daily provided gastric acid control in 15 pediatric patients with reflux esophagitis with partial clinical improvement of symptoms after 28 days of treatment. Pantoprazole was safe and well tolerated. PMID- 12548066 TI - Farber disease diagnosed after liver transplantation. PMID- 12548067 TI - Severe intestinal lymphangiectasia complicated by nephrotic syndrome treated by small bowel, liver, and kidney transplantation. PMID- 12548068 TI - Intestinal malrotation and omental cyst presenting as fetal ascites. PMID- 12548070 TI - Assessment of three methods of pH probe positioning in preterm infants. PMID- 12548069 TI - Spontaneous perforation of the bile duct in infancy: a rare but important cause of irritability and abdominal distension. PMID- 12548071 TI - Montelukast: use in pediatric patients with eosinophilic gastrointestinal disease. PMID- 12548072 TI - Death due to use of traditional medicines in Africa: a preventable cause of neonatal and infant mortality. PMID- 12548073 TI - Pharmacokinetic considerations of formulation: extended-release metoprolol succinate in the treatment of heart failure. AB - Extended-release (ER) metoprolol succinate is a controlled-release formulation designed to deliver metoprolol succinate at a near constant rate for approximately 20 h, independent of food intake and gastrointestinal pH. Once daily dosing of ER metoprolol succinate 12.5-200 mg produces even plasma concentrations over a 24-h period, without the marked peaks and troughs characteristically observed with the immediate-release (IR) formulation. This leads to consistent beta1-blockade over 24 h, while maintaining cardioselectivity at doses up to 200 mg daily. Pharmacokinetic studies have also been performed in heart failure patients and have demonstrated that ER metoprolol succinate is associated with a more pronounced and even beta1-blockade over a 24-h period than the IR formulation. The efficacy and good tolerability of ER metoprolol succinate in heart failure patients has now been demonstrated in a large-scale clinical trial. PMID- 12548074 TI - Cocaine-induced endothelin-1-dependent spasm in rabbit basilar artery in vivo. AB - Although cocaine-induced constriction of cerebral vessels may play an important negative role in the pathogenesis of cocaine-related stroke, the mechanism underlying the vasospasm remains unclear. This study investigated the role of endothelin-1 in mediating the spasm. Intracisternal cocaine infusion (10 microl/h via osmotic pump) into the cisterna magna of rabbits induced time- and concentration-dependent spasm. Maximal spasm was achieved with 100 microM cocaine infusate, and was observed as early as 0.5 days and reached a maximum at 2 days. Coinfusion of 100 microM cocaine with the endothelin receptor antagonist PD145065 (100 microM) prevented the spasm. Cerebral spinal fluid levels of cocaine and benzoylecgonine, a major cocaine metabolite, were below the limit of assay detection. This study demonstrates the novel finding that endothelin-1 mediates cocaine-induced cerebral vasospasm. PMID- 12548075 TI - Decreased modulation by lipopolysaccharide of aortic smooth muscle contractility in streptozotocin-induced hyperglycemic rats. AB - Infection is a major complication of patients with diabetes, and endotoxemic shock is a serious complication during sepsis. The purpose of this study was to determine whether the action of bacterial lipopolysaccharide (LPS) on vasocontractility is altered in diabetic vessels. Diabetes was induced in 10-week old Wistar rats by an intraperitoneal injection of streptozotocin. LPS-induced increase in cGMP (cyclic guanosine 3',5'-monophosphate) level was lower in aortae from streptozotocin-induced hyperglycemic (diabetic) rats than in those from vehicle-injected control rats, while LPS-induced nitric oxide production was not different in the diabetic and control aortae. Phenylephrine-induced contraction of diabetic aortae was lower than that of the control aortae. LPS treatment resulted in depression of contractile response to phenylephrine in both diabetic and control aortae, and the degree of depression was much lower in diabetic aortae. Treatment with N monomethyl l-arginine (l-NMMA) prevented diminution of phenylephrine-induced contraction of the aortae after LPS stimulation, and the degree of the preventive effect by l-NMMA was significantly lower in diabetic aortae than in the control aortae. Protein expression of inducible nitric oxide synthase detected by Western blot analysis was not different in the diabetic and control aortae. The decrease in cGMP production after LPS stimulation in diabetic aortae was not prevented by treatment of the aortae with superoxide dismutase but was partially prevented by that with Tiron (4,5-dihydroxy-1,3-benzene disulfonic acid), a cell-permeable scavenger of reactive oxygen species. These results suggest that LPS-induced depression of vasocontractility is attenuated in diabetic aortae due to a decrease in nitric oxide-stimulated cGMP production, probably resulting from increased inactivation of inducible nitric oxide by excessive intracellular oxidative stress. It is concluded that contractility of aortae from streptozotocin-induced hyperglycemic rats may be less affected by LPS during endotoxemia. PMID- 12548076 TI - Long-term treatment with propagermanium suppresses atherosclerosis in WHHL rabbits. AB - Macrophages play an important role in the pathogenesis of atherosclerosis, for which monocyte chemoattractant protein (MCP)-1 and CCR2 chemokine receptors may be involved. The authors have recently demonstrated that propagermanium exerts inhibitory effect on the CCR2 receptors. In the current study, the authors examined whether the organic germanium suppresses the MCP-1-induced monocyte migration in vitro and the development of atherosclerosis in WHHL rabbits in vivo. In the in vitro experiment, propagermanium concentration-dependently suppressed the MCP-1-induced migration of THP-1 cells. In the in vivo experiment, 20 WHHL rabbits were randomly divided into two groups; one group was treated with oral administration with propagermanium (9 mg/kg/day) for 3 months, and another group served as a control (n = 10 each). After 3 months, the aorta was isolated and stained with oil red O staining, and neointimal formation was quantified. Macrophage accumulation in the aorta was also evaluated by immunostaining. Long term treatment with propagermanium did not affect the serum lipid profiles. However, the treatment significantly suppressed the oil red O-positive area of the total aorta (p < 0.05). Similarly, propagermanium significantly suppressed the intimal lesions (maximal intimal thickness and intimal area) and macrophage staining-positive area (all p < 0.05). A significant positive correlation was noted between macrophage staining-positive area and intimal lesions (p < 0.0001). These results indicate that long-term treatment with propagermanium suppresses the development of atherosclerosis in WHHL rabbits, suggesting its usefulness for the treatment of atherosclerotic vascular disease in humans. PMID- 12548077 TI - Differences between angiotensin-converting enzyme inhibition and angiotensin II AT1 antagonism on angiotensin-mediated responses in human internal mammary arteries. AB - The current study aimed to demonstrate differences between angiotensin (Ang) converting enzyme (ACE) inhibition and Ang II-AT1 receptor antagonism on full concentration-contraction responses to Ang I. Contraction responses to increasing concentrations of Ang I (1 nM-1 microM) were evaluated in organ baths in the presence of captopril (10 microM-1 mM) with or without a chymase inhibitor (1 microM soybean trypsin inhibitor), or irbesartan (0.1 nM-microM), in internal mammary arteries from 25 patients undergoing coronary bypass surgery. Responses were expressed as a percentage of the control response to 10 microM phenylephrine. Captopril did not change the maximum response to Ang I (control: 46.3 +/- 6.3%, captopril: 43.0 +/- 4.6%). In contrast, 0.1 microM irbesartan completely blocked the maximum response to Ang I (from 45.8 +/- 6.7% to 1.9 +/- 1.9%, p < 0.001). However, addition of soybean trypsin inhibitor to captopril more effectively shifted -log pD2 than captopril alone (0.47 +/- 0.06 vs 0.95 +/- 0.14 log units, p = 0.007). Ang I-mediated effects are much more effectively inhibited by Ang II antagonism than by ACE inhibition. The incomplete effects of captopril on the inhibition of Ang II formation might be caused by alternative Ang II forming enzyme(s), as was demonstrated by the additive effects of soybean trypsin inhibitor added to captopril. PMID- 12548078 TI - Effect of folic acid on nitrate tolerance in healthy volunteers: differences between arterial and venous circulation. AB - This study investigated whether oral supplemental folic acid can prevent the development of nitrate tolerance and whether it has different effects on the arterial and venous systems. Twenty-four healthy male volunteers received either placebo or folic acid (10 mg/d) for 14 days. Additionally, all subjects underwent concurrent transdermal nitroglycerin therapy for 7 days. Venous occlusion forearm strain gauge plethysmography measured arterial and venous responses to sublingual nitroglycerin before and after treatment. Both arterial and venous responses were blunted in the placebo group after transdermal nitroglycerin. Folic acid prevented the development of nitrate tolerance in arteries but had no effect in veins. PMID- 12548079 TI - Electrophysiologic characterization of dronedarone in guinea pig ventricular cells. AB - The electrophysiological properties of dronedarone (SR33589), a noniodinated amiodarone-like agent, were studied on action potential (AP) and contraction of papillary muscle and on membrane ionic currents, Ca2+ transient, and shortening of ventricular cells of the guinea pig heart. In multicellular preparations, dronedarone (3, 10, and 30 microM) decreased maximum rate of rise of AP (dV/dt max) with a concentration- and frequency-dependent relationship; resting potential was not modified and AP amplitude was decreased only at 30 microM. The effects of dronedarone on AP durations (APDs) at different percentages of repolarization were not significantly changed, except for a slight decrease in APD30 and APD50 at the highest concentration. In isolated ventricular myocytes, dronedarone inhibited rapidly activating delayed-rectifier K+ current (I(Kr)) (median inhibitory concentration [IC50] /= 30 microM). Dronedarone blocked L type Ca2+ current (I(Ca(L))) (IC50 = 0.18 +/- 0.018 microM at a stimulation frequency of 0.033 Hz) in a use- and frequency-dependent manner. Simultaneously to these electrophysiological effects, dronedarone reduced contraction amplitudes of papillary muscle and decreased Ca2+ transient and shortening of ventricular myocytes. The results show that dronedarone is a multichannel blocker because it decreases dV/dt max (I(Na)), I(Ca(L)), I(Kr), I(Ks), and I(K1). These effects are accompanied by a reduction in free intracellular calcium and contraction amplitudes. Dronedarone does not significantly change APD whatever the stimulation frequency. Our data demonstrate that the acute electrophysiological characteristics of dronedarone, despite absence of iodine in its molecular structure, are very similar to those of amiodarone in cardiac ventricle. PMID- 12548080 TI - Hemodynamic and cardiac anti-hypertrophic actions of clonidine in Goldblatt one kidney, one-clip rats. AB - In congestive heart failure, the chronic sympathetic hyperactivity contributes to a poor prognosis. In this respect, clonidine, a centrally acting sympathoinhibitory drug, has previously been tested in clinical trials. The aim of the current study was to evaluate the effects of clonidine on morbidity and mortality in an experimental model of cardiac hypertrophy associated with hypertension, renal failure, and intense sympathetic activation. One-kidney, one clip Goldblatt hypertensive rats were treated orally with clonidine (200 microg/kg/d) during 30 days and were compared with untreated rats and with sham operated animals. Cardiac hemodynamics, left ventricular volume and elasticity, cardiac morphometry and histology, and renal function were evaluated. A survival study was also performed. Clonidine normalized cardiac function, ventricular stiffness, and prevented ventricular structural remodeling. Moreover, despite a marked renal function impairment, survival of the animals was increased in the clonidine group. The centrally acting sympathoinhibitory drug clonidine exhibited marked cardioprotective properties. This study emphasized the interest of evaluating drugs whose aim is to treat congestive heart failure, in an experimental model of cardiac and renal failure. PMID- 12548081 TI - Influence of L-nitro-arginine methyl ester, acetylcholine, and adenosine on mean blood pressure, pulse pressure, and pulse pressure amplification in rats. AB - The blood pressure pattern in spontaneously hypertensive rats (SHRs) involves three main characteristics: increase in mean blood pressure (MBP); increase in thoracic aorta (proximal) and iliac (distal) pulse pressure (PP); disappearance of the normal PP amplification between the proximal and the distal arteries. Whether pharmacologic agents may reduce MBP with different or even opposite effects regarding PP and PP amplification has been poorly investigated. In SHRs and Wistar-Kyoto (WKY) anesthetized rats, the NO inhibitor l-nitro-arginine methyl ester (l-NAME) was infused at the dosage of 1 mg/kg for 30 min. Before and after infusion, 7 microg/kg/min acetylcholine (Ach) and 200 mg/kg adenosine (Ado) were perfused for 4 min. Proximal and distal intra-arterial BP was monitored throughout the procedure. In both WKYs and SHRs, l-NAME increased proximal and distal systolic (SBP), diastolic (DBP), and MBP but not PP. Before l-NAME, SBP, DBP, and MBP were significantly reduced by Ado and Ach. After l-NAME, such blood pressure reductions were abolished with Ach but not Ado. In both strains, the proximal and distal PP, when expressed in percent reduction of MBP, were significantly higher under Ado than under Ach. The Ado but not Ach changed PP amplification, causing a reduction in WKYs and an increase in SHRs independent of l-NAME. Vasodilating agents may reduce MBP with significantly different effects on PP. The Ado alters PP amplification, an effect not obtained with the nitric oxide endothelium-dependent vasorelaxing agent Ach. Tail SBP measurements cannot predict such dissociated changes. PMID- 12548082 TI - Effect of high-dose statin treatment on plasma concentrations of endogenous nitric oxide synthase inhibitors. AB - Asymmetric dimethylarginine (ADMA) and monomethylarginine (LMMA) are endogenous inhibitors of nitric oxide synthase. A high level of ADMA in plasma has shown to be a significant risk factor for acute coronary syndromes and elevated plasma ADMA levels are prevalent in patients with hypercholesterolemia. It was therefore hypothesized that lowering plasma cholesterol levels with statin treatment would also lower ADMA concentrations. This double-blind study addressed the effect of high-dose statin treatment on plasma levels of ADMA and LMMA. Forty-eight subjects with mild hypercholesterolemia were randomly assigned to receive simvastatin 80 mg/d, atorvastatin 40 mg/d, or placebo for 8 weeks. Both statins decreased low-density lipoprotein cholesterol effectively (simvastatin 54% and atorvastatin 49%). However, concentrations of arginine derivatives remained unchanged during statin treatment and did not correlate with cholesterol levels. This study indicates that statin treatment has no clear influence on plasma ADMA or LMMA concentrations. PMID- 12548083 TI - Commonly used numbers of microspheres affect cardiac vascular resistance. AB - The main goal of the study was to examine how the microsphere technique affects the hemodynamics and mitochondrial energy status of the Langendorff-perfused rat heart. The hearts were perfused at a constant flow with Tyrode solution. NADH videofluorometry of the surface of the left ventricle was used to record the mitochondrial energy status as indication of regional ischemia. The effects of seven successive (separated by 10 min) injections of 0.1 ml of saline or (0.05% Tween 20; polysorbate 20, Sigma-Aldrich, St. Louis, MO, U.S.A.) or (0.05% Tween 20 + microspheres) were studied. The number of microspheres per injection were: #1 (2,500), #2 (5,000), #3 (10,000), #4 (20,000), #5 (40,000), #6 (40000), and #7 (80000). The anti-aggregation agent Tween always caused a biphasic response in perfusion pressure. Compared with the Tween effect, the injection of microspheres caused an initial change (mm Hg) in perfusion pressure of #1 (-10), #2 (NS), #3 (NS), #4 (+7.5), #5 (+12.3), #6 (+14.4), #7 (+18.3), and a delayed change (10 min after injection) of #1 (-22.2), #2 (-6.0), #3 (-4.1), #4 (-4.5), #5 (NS), #6 (NS), and #7 (+5.9). The microspheres caused a significant delayed increase in NADH only for injection #6 and #7. Similar results were found for different durations of the input function or when hearts were perfused at constant perfusion pressure. In hearts without flow reserve (10 microM adenosine), Tween injections were without effect, whereas three successive injections of 60,000 microspheres each only caused increases in perfusion pressure and NADH. The data demonstrate that in hearts with flow reserve present, even very low numbers of microspheres (2,000/g heart) cause large decreases in perfusion pressure without obvious signs of ischemia. When flow reserve was exhausted by either microsphere loading or adenosine addition, microspheres only caused increases in perfusion pressure and resulted in detectable ischemia (NADH). It is concluded that microspheres affect the vascular resistance of the heart and that these effects are flow reserve dependent. PMID- 12548084 TI - Effects of long-term treatment with ketanserin on blood pressure variability and end-organ damage in spontaneously hypertensive rats. AB - It has been proposed that instability of blood pressure may produce organ damage. Ketanserin is an anti-hypertensive drug with an ability to reduce blood pressure variability (BPV) in acute experiments in spontaneously hypertensive rats (SHRs). The present work was designed to observe the effects of long-term treatment with ketanserin on BPV and end-organ damage in SHRs. Ketanserin was mixed in rat chow at an estimated dose of 10 mg/kg/d. After 5 months of drug administration, BP was continuously recorded in conscious, freely moving rats for 24 h. The heart, kidneys, and abdominal aorta were then isolated and examined by using histologic methods and computer image analysis. In another work, the effects of hydralazine (40 mg/kg/d, for 5 months) on BP, BPV, and organ damage were observed in SHRs. Ketanserin significantly decreased BP and BPV, ameliorated impaired arterial baroreflex function, and significantly prevented the target organs of SHRs from being damaged. This preventive effect was characterized by decrease in left ventricular hypertrophy, diminution of glomerulus damage, and amelioration in vascular lesion. Hydralazine decreased BP but did not lower BPV. No organ protection was found in hydralazine-treated rats. In conclusion, long-term treatment with ketanserin reduced hypertensive organ damage. Lowering BP, decreasing BPV, and ameliorating arterial baroreflex function may contribute together to this effect. PMID- 12548085 TI - 8'-aminoguanosine inclusion results in enhanced efflux of taurine in preconditioned ischemic myocardium. AB - Attenuated purine levels are characteristic findings of ischemic preconditioning (PC). Lower energy demand in PC myocardium leading to less nucleotide decay is a reasonable explanation. However, experimental data suggest that the activities of the enzymes involved in purine metabolism are increased in PC myocardium. Recently it was suggested that PC favored degradation of exogenous adenosine to inosine successively ending up in enhanced lactate production. This was probably because of the involvement of the hexose monophosphate pathway in the PC ischemic myocardium. This route may therefore be supplementary in energy metabolism as a metabolic flow can be started by adenosine ending up in lactate without initial adenosine 5'-triphosphate (ATP) investment. Purine nucleoside phosphorylase (PNP) is a key enzyme in the proposed metabolic route. In the current study the effect of PNP inhibition (with 8'-aminoguanosine) on myocardial energy metabolism during PC was studied in an open chest porcine heart model using the microdialysis technique. A dose-dependent inhibition of PNP by 8'-aminoguanosine was observed in PC myocardium. This inhibition resulted in an enhanced exodus of taurine reflecting a disturbed energy economy of the cardiomyocytes. Addition of inosine being a true substrate of PNP reversed these changes, which indicated that 8' aminoguanosine was a competitive inhibitor of PNP. It is concluded that the ischemic PC phenomenon at least partly involves the activated enzyme PNP. PMID- 12548086 TI - Prostaglandin E1 improves endothelial function in critical limb ischemia. AB - Prostaglandin E1 (PGE1) may relieve rest pain and heal ulcers in critical limb ischemia, but its mechanism of action is still incompletely understood. To investigate the effects of PGE1 treatment on endothelial function evaluated as brachial artery flow-mediated vasodilation (FMV) and on soluble adhesion molecule plasma levels (vascular adhesion molecule-1 [sVCAM-1] and intercellular adhesion molecule-1 [sICAM-1]), 12 patients with critical limb ischemia were treated with daily PGE IV infusion (alprostadil 60 microg) for 2 weeks. FMV and plasma sICAM-1 and sVCAM-1 concentrations were determined at baseline, after the first infusion, and after 1 and 2 weeks. Compared with 30 healthy control subjects, patients had higher baseline sVCAM-1 (2.402 +/- 296 ng/ml vs 972 +/- 117 ng/ml) and sICAM-1 levels (464 +/- 51 ng/ml vs 206 +/- 37 ng/ml, both p < 0.05) and lower FMV (1.0 +/- 1.1% vs 5.6 +/- 1.6%, p < 0.05). sICAM-1 concentration progressively decreased with treatment (from 464 +/- 51 ng/ml to 326 +/- 56 ng/ml, 288 +/- 42 ng/ml, and 279 +/- 44 ng/ml after the first dose and, respectively, after 1 and 2 weeks; all p < 0.05). sVCAM-1 showed a reduction after 2 weeks (from 2.402 +/- 296 ng/ml to 1.916 +/- 176 ng/ml; p < 0.05). FMV improved after 1 and 2 weeks (from 1.0 +/- 1.1% to 3.1 +/- 0.6% and 5.2 +/- 2.1%, both p < 0.05). In conclusion, treatment with PGE1 determines a significant improvement in endothelial function in patients with critical limb ischemia. PMID- 12548087 TI - Effects of dual angiotensin-converting enzyme and neutral endopeptidase 24-11 chronic inhibition by mixanpril on insulin sensitivity in lean and obese Zucker rats. AB - The aim of this study was to examine the effects of chronic (8-day) oral treatment with the dual angiotensin-converting enzyme (ACE) and neutral endopeptidase 24-11 (NEP) inhibitor mixanpril (25 mg/kg twice a day), compared with the ACE inhibitor captopril (25 mg/kg twice a day), on whole body insulin mediated glucose disposal in young (10-week) and old (19-week) obese Zucker rats (ZOs). Moreover, the effects of chronic mixanpril administration on femoral blood flow at rest and during an insulin infusion were assessed. In the young ZOs, mixanpril decreased the glucose response during an IV glucose tolerance test more effectively than did captopril (-49 and -30%, respectively, p < 0.05). Incremental glucose area under the curve in mixanpril-treated ZOs was then no longer different from that observed in vehicle-treated lean rats (1,592 +/- 175 and 1, 470 +/- 104 mg/dl x min, respectively). The beneficial effects resulting from mixanpril or captopril administration were observed in ZOs but not in lean littermates. In the old ZOs, mixanpril induced higher glucose infusion rates to maintain euglycemia than did captopril during a hyperinsulinemic euglycemic clamp test (+92 and +35%, respectively, p < 0.001). However, the glucose infusion rates in mixanpril-treated ZOs remained much lower than that observed in vehicle treated lean rats (9.4 +/- 0.7 mg/kg/min vs 28.6 +/- 1.0 mg/kg/min, p < 0.001). Mixanpril did not affect resting femoral vascular bed hemodynamics but restored the femoral blood flow response to insulin infusion. In conclusion, in ZOs, chronic dual ACE/NEP inhibition improves whole body insulin-mediated glucose disposal more effectively than does ACE inhibition alone. This beneficial effect seems to be restricted to conditions of insulin resistance and not directly linked to the improvement in the femoral blood flow response to insulin. PMID- 12548088 TI - Possible involvement of facilitated polyol pathway in augmentation of intimal hyperplasia in rabbits with alloxan-induced hyperglycemia. AB - Present experiments were designed to investigate whether the facilitated polyol pathway is involved in the augmentation of intimal hyperplasia with hyperglycemia. Twelve weeks after a single bolus intravenous injection of alloxan (100 mg/kg) or saline, rabbits underwent a unilateral endothelial denudation of the carotid artery. Intimal hyperplasia was evident 4 weeks after denudation and significantly augmented in hyperglycemic animals treated with alloxan. This effect was accompanied by the enhanced accumulation of endogenous NOS inhibitors (N(G)-monomethyl-l-arginine [l-NMMA] and asymmetric, N(G),N(G)-dimethyl-l arginine [ADMA]) in regenerated endothelial cells, impairment of NO production and release, and enhanced accumulation of endothelin-1 (ET-1) within the vessel wall. Sorbitol levels in aortic endothelial cells and within the smooth muscle layer were significantly increased with hyperglycemia. All these changes associated with hyperglycemia were significantly reduced in animals treated with the selective aldose reductase inhibitor fidarestat (3 mg/kg/d). These findings suggest that the facilitated polyol pathway possibly plays an important role for the augmentation of intimal hyperplasia caused by the hyperglycemic state. PMID- 12548089 TI - Angiotensin-(1-7) reduces renal angiotensin II receptors through a cyclooxygenase dependent mechanism. AB - In the kidney, angiotensin-(1-7) [Ang-(1-7)] exhibits diuretic and natriuretic properties associated with an increase in prostaglandin production. The prohypertensive effects of Ang II are attenuated in rats infused with Ang-(1-7), consistent with recent work showing that Ang-(1-7) downregulates AT1 receptors in Chinese hamster ovary-AT1A or vascular smooth muscle cells. To determine whether exposure to Ang-(1-7) reduces AT1 receptors in the kidney through an increase in prostaglandin production, kidney slices from Sprague-Dawley rats were incubated with 10 n -1 microM Ang-(1-7) in the presence or absence of 5 microM meclofenamate, a cyclooxygenase inhibitor. Following these treatments, the kidney slices were retrieved, frozen, and sectioned for determination of [125I]-Ang II binding using in vitro receptor autoradiography. Greater than 90% of the specific binding was competed for by losartan, indicating that the majority of binding was to the AT1 receptor. Incubation of kidney slices with 1 microM Ang-(1-7) caused a 20% reduction in [125I]-Ang II binding (n = 8) in the cortical tubulointerstitium, which was prevented when Ang-(1-7)-treated slices were incubated in the presence of 5 microM meclofenamate (1 +/- 2% increase, n = 8; p < 0.05). Incubation with 5 microM meclofenamate alone had no effect on [125I]-Ang II binding (-3 +/- 3%). The decrease in [125I]-Ang II binding with Ang-(1-7) was also blocked by the Ang-(1-7) antagonist [d-Ala7]-Ang-(1-7). Treatment with 1 microM [d-Ala7]-Ang-(1-7) alone had no effect on [125I]-Ang II binding (-3 +/- 6% of control). Pretreatment with 1 microM Ang II caused a similar reduction in [125I]-Ang II binding in the cortical tubulointerstitium. Neither Ang-(1-7) nor Ang II had any effect on [125I]-Ang II binding in the glomeruli and the area of the vasa recta of the kidney. These original findings suggest that prior exposure to Ang-(1-7) or Ang II causes a modest decrease in the number of AT1 receptors in the cortical tubulointerstitial area of the kidney. The reduction in Ang II binding by Ang-(1-7) was blocked by meclofenamate and [d-Ala7]-Ang-(1-7), suggesting that cyclooxygenase products released through activation of a novel receptor participate in this effect. PMID- 12548090 TI - Lack of oxidative stress during sustained therapy with isosorbide dinitrate and pentaerythrityl tetranitrate in healthy humans: a randomized, double-blind crossover study. AB - The mechanisms by which tolerance to organic nitrates develops are still poorly understood. Enhanced oxidative stress, i.e., increased free radical production following organic nitrate administration, has been recently suggested as a possible mechanism. A randomized, double-blind, crossover study assessed in 18 healthy young volunteers at baseline and 1 and 5 days after oral administration with therapeutically relevant doses of isosorbide dinitrate (ISDN, 30 mg TID) or pentaerythrityl tetranitrate (PETN, 80 mg TID) the effect on two index parameters of oxidative stress in vivo, i.e., urinary 8-iso-prostaglandin (PG)F2alpha and circulating 3-nitrotyrosine and their major urinary metabolites, 2,3-dinor-5,6 dihydro-8-iso-PGF2alpha and 3-nitro-4-hydroxyphenylacetic acid. In addition, urinary cGMP and serum and urinary nitrate and nitrite were determined. All parameters were quantified by gas chromatography-mass spectrometry or gas chromatography-tandem mass spectrometry except for cGMP, which was analyzed by radioimmunoassay. Serum and urinary nitrite levels increased significantly following 5-day administration of ISDN and PETN. Neither urinary excretion of 8 iso-PGF2alpha and plasma 3-nitrotyrosine nor their respective metabolites changed significantly after ISDN or PETN administration. There were no significant differences between ISDN and PETN regarding these parameters. Urinary cGMP increased significantly only after ISDN. This study is compatible with a stimulation of cGMP by ISDN, but neither ISDN nor PETN enhances systemic oxidative stress in healthy volunteers. PMID- 12548091 TI - Efficacy and safety of treprostinil: an epoprostenol analog for primary pulmonary hypertension. AB - Intravenous epoprostenol is currently FDA approved for management of primary pulmonary hypertension, but it requires intravenous infusion and is associated with adverse effects. The objective of this study was to evaluate the effects of an epoprostenol analog, treprostinil, for management of pulmonary hypertension. Ten tertiary care academic institutions with pulmonary hypertension programs participated in these pilot trials. In the first trial, intravenous epoprostenol and intravenous treprostinil were compared. In the second trial, intravenous treprostinil and subcutaneous treprostinil were compared. In the third trial, subcutaneous treprostinil was compared with placebo infusion during an 8-week period. Intravenous epoprostenol and intravenous treprostinil resulted in a similar reduction in pulmonary vascular resistance acutely (22% and 20%, respectively). Intravenous treprostinil and subcutaneous treprostinil also demonstrated comparable short-term decrease in pulmonary vascular resistance (23% and 28%, respectively). The placebo-controlled 8-week trial demonstrated a mean improvement of 37 +/- 17 m as measured by the 6-minute walk distance in patients receiving treprostinil compared with a 6 +/- 28 m reduction in those receiving placebo. There were trends toward an improvement in cardiac index and pulmonary vascular resistance index in the treprostinil group. Subcutaneous treprostinil has favorable hemodynamic effects when given acutely and in the short term. Treprostinil can be given safely to an ambulatory patient with a novel subcutaneous delivery pump system. PMID- 12548092 TI - Role of nitric oxide and K+ channels in relaxation induced by polygodial in rabbit corpus cavernosum in vitro. AB - This study examines the relaxation produced by the sesquiterpene polygodial and compares its action with those caused by acetylcholine (ACh) and sodium nitroprusside (SNP) in the rabbit corpus cavernosum (RbCC) in vitro. RbCC was set up in a 5-ml bath containing Krebs solution at 37 degrees C, at pH 7.2, and under 2 g of tension. Polygodial, ACh, and SNP elicited graded relaxation in RbCC with mean EC50 values of 46.70 microM, 0.38 microM, and 0.30 microM, respectively. The nitric oxide (NO) synthase inhibitor L-NOARG and the guanylate cyclase inhibitors LY 83583 and ODQ markedly inhibited the relaxation induced by polygodial (% of inhibition of 79, 48, and 51, respectively) and those caused by ACh (% of inhibition of 100, 49, and 32, respectively). Tetraethylammonium (TEA) and glibenclamide inhibited the relaxation induced by polygodial (52% and 43%, respectively), but only TEA caused shift to the right on ACh-mediated relaxation. In contrast, apamin, charybdotoxin, and 4-aminopyridine or the protein kinase A inhibitor KT 5720 all failed to affect either polygodial or ACh-mediated relaxation in these preparations. The authors concluded that polygodial produced graded relaxation in the RbCC in vitro via a mechanism that was partially dependent on the release of NO or a NO-derived substance through an activation of guanylate cyclase but was independent of adenylate cyclase mechanism. In addition, the opening of K+ channels sensitive to TEA and glibenclamide, but not those sensitive to apamin, 4-aminopyridine, or charybdotoxin, also contributed to the relaxant action produced by polygodial in the RbCC. PMID- 12548093 TI - Pharmacologic characterization of contractile serotonergic receptors in human isolated mesenteric artery. AB - The 5-hydroxytryptamine (5-HT) receptors mediating contraction in human isolated mesenteric arteries were characterized. Endothelium-denuded human isolated mesenteric arteries were used. 5-HT induced concentration-dependent contractions in mesenteric arteries (Emax, 127.37 +/- 7.61% of 80 mM KCl maximal contraction; pD2, 6.73 +/- 0.09 [-logEC50]). Sumatriptan, a selective 5-HT1B/1D receptor agonist, induced concentration-dependent contractions in some of the arteries (Emax, 61.82 +/- 10.04%; pD2, 6.56 +/- 0.21, n = 9) but not in the others (Emax < 5%, n = 13), suggesting that functional 5-HT1B/1D receptors exist in some but not in all mesenteric arteries. GR127935 (a selective 5-HT1B/1D receptor antagonist, 3 nM) inhibited sumatriptan-induced contractions in arteries in which sumatriptan responses were strong in an insurmountable manner. GR127935 (10 nM) also inhibited 5-HT responses and shifted the concentration-response curve of 5-HT to the right significantly (p < 0.05; pD2s were 6.54 +/- 0.18 and 5.93 +/- 0.11 in the presence of vehicle and GR127935, respectively). Ketanserin (0.01-1 microM) competitively antagonized 5-HT responses in human mesenteric arteries: pA2 value was 8.40 +/- 0.25 (slope of Schild regression, 1.43 +/- 0.18; r2, 0.98). Tropisetron (5-HT3 receptor antagonist) and prazosin (alpha1-adrenoceptor antagonist) did not affect the contractions induced by 5-HT. These results suggest that 5-HT2A and 5-HT1B/1D receptors, but not 5-HT3 and alpha1 adrenoceptors, are involved in the 5-HT-induced contractions in human isolated mesenteric arteries. Sumatriptan-induced and 5-HT1B/1D receptor-mediated responses vary greatly among patients. PMID- 12548094 TI - Effect of atorvastatin on adhesive phenotype of human endothelial cells activated by tumor necrosis factor alpha. AB - We studied the effect of atorvastatin on the adhesive phenotype of human endothelial cells (HUVEC) stimulated by tumor necrosis factor (TNF)-alpha. Surface expression of adhesion molecules on HUVEC was examined by flow cytometry and confocal microscopy, and adhesion of monocytes (human THP-1 cell line) was measured in vitro under flow conditions. In TNF-alpha-activated HUVEC, atorvastatin significantly enhanced surface expression of vascular cell adhesion molecule (VCAM)-1, intercellular adhesion molecule (ICAM)-1, E-selectin, and fractalkine, when compared with TNF-alpha stimulation alone. This enhancement was reversed by mevalonate or geranylgeranyl pyrophosphate (GGPP) and was mimicked by an inhibitor of geranylgeranylation. The enhancing effect of atorvastatin was restricted to TNF-alpha-inducible adhesion molecule and was the reflect of an increased protein synthesis (mRNA and protein) and not of a reduced shedding. Confocal microscopy examination showed that atorvastatin also altered the surface distribution of adhesion molecules. Adhesion of human THP-1 cells on TNF-alpha activated HUVEC was significantly reduced by atorvastatin (-42% at 1 microM). Mevalonate or GGPP restored the TNF-alpha-induced adhesive potential. These results show that atorvastatin, by inhibiting prenylation of G proteins, enhances the TNF-alpha-induced expression of adhesion molecules at the endothelial cell surface and also alters their surface distribution which may account for the reduced binding of monocytes. PMID- 12548095 TI - Effects of long-term treatment with candesartan on organ damages in sinoaortic denervated rats. AB - The study was designed to observe the effects of long-term treatment with candesartan cilexetil (candesartan) on blood pressure (BP), blood pressure variability (BPV), baroreflex sensitivity (BRS) and end-organ damage (EOD) in sinoaortic denervated (SAD) rats. Candesartan was mixed in rat chow at an estimated dose of 3 mg/kg/day. After 12 weeks of drug administration, rats were instrumented to determine BP, BPV and BRS in conscious state. Organ damage was estimated by observation of morphologic changes. When compared with sham-operated rats, SAD rats exhibited increased BPV, decreased BRS, and normal BP and plasma angiotensin II level. Left ventricular and aortic hypertrophies and renal lesion were found in SAD rats. Candesartan significantly decreased BP and BPV, ameliorated impaired BRS, increased plasma angiotensin II level and obviously diminished the EOD in SAD rats. Multiple-regression analysis shows that decrease in left ventricular hypertrophy was mainly related to decrease in systolic BPV. Decrease in aortic hypertrophy was mainly determined by increase in BRS and decrease in systolic BP. Amelioration in renal lesion was predicted by increase in BRS and decrease in systolic BPV. BRS was the most important determinant for renal lesion and aortic hypertrophy in SAD rats. In addition, plasma angiotensin II level was higher in candesartan-treated rats. In conclusion, long-term treatment with candesartan prevented SAD-induced organ damage. Restoration of arterial baroreflex function, decrease in BPV, and blockade of activated renin angiotensin system may contribute to the organ protective action of candesartan in SAD rats. PMID- 12548096 TI - Smallpox as a biological weapon: implications for the critical care clinician. AB - Once believed eradicated, smallpox has returned as a potential threat. Healthcare providers, as the first line of defense, must be proactive in maintaining current resources and offering input into the preparation for, diagnosis, and management of smallpox should an outbreak occur. The critical care nurse is integral in all aspects regarding smallpox and bioterrorism. PMID- 12548097 TI - Electrophysiologic testing. AB - The purpose of this article is to provide an introduction to electrophysiology (EP) testing. The history and evolution of EP testing will be reviewed briefly. The indications for electrophysiology studies (EPS) testing, EP definitions and terminology, components of a basic EP procedure, and potential complications will also be presented. To understand the principles of EPS, comprehension of the heart's electrical properties is imperative. PMID- 12548099 TI - Writing for publication: you can do it! AB - Writing for publication can be an intimidating process but it does not have to be. The thrill of seeing your first publication is one you will never forget. Many nurses, for a variety of reasons, do not write articles for publication. This article will discuss the barriers to writing and how to overcome them. PMID- 12548098 TI - New thoughts on sepsis: the unifier of critical care. AB - In the United States, more than $16 billion annually is spent managing patients with severe sepsis and its sequelae. Insight into the inflammatory response, endothelial tissue, and the coagulation cascade suggest promising new treatment regimens that limit morbidity and mortality due to sepsis and multisystem organ failure. This article will discuss new information regarding the pathophysiology of the inflammatory response and sepsis. Current thoughts on clinical management and a promising new agent, Activated Protein C, will be presented. PMID- 12548100 TI - Palliative care services. AB - This article presents a program developed at one facility to help provide palliative care services to patients in the intensive care units. PMID- 12548102 TI - A closing word: critical care visitation. PMID- 12548101 TI - Decreased cardiac output: validation with postoperative heart surgery patients. AB - This study was done to analyze the association between defining characteristics of decreased cardiac output (DCO) identified by noninvasive methods and the cardiac index (CI) estimated by thermodilution. Two defining characteristics were associated with DCO: decreased peripheral pulse and decreased peripheral perfusion. This study points out the relevance of peripheral circulation assessment in postoperative cardiac surgery patients. PMID- 12548103 TI - Oral contraceptives and cervical cancer: public health implications. PMID- 12548104 TI - Hormone replacement therapy and cancer: an update. PMID- 12548105 TI - Epidemiological research on cancer with a focus on Europe. AB - Cancer has been a growing public health problem throughout the last century. Cancer mortality rates in both sexes combined in the European Union (EU) reached a peak in 1988. Thereafter, they declined by 9.4% in 1998. Likewise, lung cancer rates in the EU for both sexes combined increased by 58% between 1960 and 1988, but declined by 14% in 1998. Over a third of the decline was accounted for by lung cancer alone and approximately half by the combination of tobacco-related neoplasms. About half of the decline in total cancer mortality not attributable to tobacco derived from the steady fall in mortality from gastric cancer. The remaining half, including favourable trends in colorectal, breast, testis and lymphoid neoplasms, can be at least in part attributed to advancements in cancer diagnosis and treatment. The major causes of cancer and hence the most important priorities for research will be reviewed, with a specific focus on European priorities for research. PMID- 12548106 TI - Risk factors for gallbladder cancer: a reappraisal. AB - Carcinoma of the gallbladder has a very unusual geographical distribution with pockets of high incidence seen in Chile, Poland, India, Japan and Israel; it occurs rarely in the rest of the world. It is a common malignancy in the Western Bihar and Eastern Uttar Pradesh regions of India. Patients present with extremes of clinical symptoms, indicating benign biliary diseases on the one hand and incurable malignant disease on the other. Laboratory and roentgenographic data tend to confirm the clinical diagnosis of the advanced incurable disease at presentation, in most cases. Various aetiopathological agents have been proposed but none has stood the test of time. In this article, we have reviewed the aetiopathological agents proposed from time to time over the past two centuries, with special emphasis on the developments made in the last 25 years. PMID- 12548107 TI - Smoking, alcohol, diet, dentition and sexual practices in the epidemiology of oral cancer in Poland. AB - The effect of smoking, drinking, diet, dental care and sexual habits on the risk of oral and pharyngeal cancer was investigated in a case-control study conducted in Warsaw, Poland. The study comprised 122 patients (including 44 females) aged 23-80 years with histologically confirmed cancer of oral cavity and pharynx. Controls were 124 subjects (including 52 females) admitted to the hospital for different non-neoplastic conditions unrelated to tobacco and alcohol consumption, with frequency matched to cases by age and sex. Smoking and drinking were strongly associated with an increased risk of oral cancer. Among consumers of both products, risks of oral cancer tended to combine in a multiplicative fashion and were increased more than 14-fold among those who consumed more than 15 cigarettes and seven or more drinks per day. Cessation of smoking was associated with reduced risk of this cancer. The risks varied by type of cigarettes smoked, being lower among those consuming filtered cigarettes only (OR = 1.6) than nonfilter (OR = 6.5) or mixed (OR = 4.2) cigarettes. High fruit intake was associated with significantly decreased risk (OR = 0.4) with the strongest significant inverse association found for fruit juices and citrus fruits ( < 0.01). After adjustment for tobacco smoking and alcohol drinking, poor dentition as evidenced by missing teeth, frequency of dental check-ups and frequency of teeth brushing emerged as a strong risk factor. Number of missing teeth and frequency of dental check-ups and frequency of tooth brushing showed increased ORs of 9.8, 11.9 and 3.2, respectively. Denture wearing did not affect oral cancer risk. No differences were detected in sexual practices (including oral sex and intercourse with prostitutes). In terms of attributable risk, smoking accounted for 57% of oral cancer cases in Poland, alcohol for 31% and low fruit intake for 12%. Attributable risks for low frequency of tooth brushing and dental check-ups were 56% and 47%, respectively. In conclusion, smoking and drinking cessation and increase of fresh fruit intake are likely to be effective preventive measures against oral cancer. These findings indicate also that poor oral hygiene may be an independent risk factor. PMID- 12548109 TI - Cutaneous melanoma in the Florentine area, Italy: incidence, survival and mortality between 1985 and 1994. AB - In recent decades, the increase in incidence of melanoma (MM) and the consequent mortality pointed to the concept of a 'melanoma epidemic'. More recently, the mortality has been slowly declining in many countries. This study is aimed at evaluating the incidence, mortality and survival in the Florentine area of Italy, using registry-based information. Between 1985 and 1994, 997 cases were notified with a survival of 713 patients (1985-92) and 316 deaths. Age-adjusted incidence, mortality rates and 95% confidence interval were calculated by period, gender and Breslow thickness. The relative survival rates were calculated and the effects of prognostic factors were evaluated using multivariate analysis. The incidence of MM increased during this period. This result referred only to 'thin melanomas', while the incidence rate for melanomas thicker than 1.00 mm remained unchanged both in young and old individuals. The mortality rate remained stable. The 5-year survival rate increased between 1985 and 1992. The inclusion of Breslow thickness in the multivariate model caused a reduction of the period effect. In conclusion, a changing pattern of MM epidemiology is being observed, with increase of 'thin' forms and a tendency for mortality to decline. The increasing awareness of population about MM prevention may partially explain these findings. PMID- 12548108 TI - Maternal and gestational correlates of pregnancy prolactin and growth hormone in USA and China. AB - The objective of this study is to determine correlates of prolactin and growth hormone levels among pregnant women in the USA and China. We studied 304 pregnant Caucasian and 335 pregnant Chinese women. Levels of prolactin and growth hormone were measured at weeks 16 and 27 of gestation, and correlated with maternal, gestational and perinatal characteristics. Both growth hormone and, to a lesser extent, prolactin were inversely associated with pregnancy weight and body mass index, history of a previous live birth and newborn size, whereas educated women had higher levels of both hormones. Growth hormone levels were lower in women who gained more weight, smoked and had nausea and vomiting during pregnancy, whereas prolactin increased with longer total gestation. We found robust associations between maternal and newborn characteristics on the one hand and prolactin and growth hormone during pregnancy on the other. PMID- 12548110 TI - Oral and oropharyngeal cancer in Spain: influence of dietary patterns. AB - Cancer of the oral cavity and oropharynx is a major public health problem in Spain. Tobacco and alcohol have been identified as the two major risk factors for oral cancer in most western populations. Other risk factors include diets low in fruits and vegetables, but the impact of dietary habits on the risk of these cancers has never been assessed in Spain. The objective of this study was to elucidate the role of dietary habits in oral and oropharyngeal cancer, a multicentric case-control study was conducted in three areas of Spain (Barcelona, Granada and Sevilla) between 1996 and 1999. Cases were 375 patients (71 women), with incident, histologically confirmed cancer of the oral cavity or oropharynx, and controls were 375 subjects (71 women) admitted to hospitals for conditions unrelated to smoking or alcohol drinking. Data were analysed using multivariate logistic regression procedures. After allowance for education, tobacco and alcohol use, a significant inverse association with the risk of oral and oropharyngeal cancer was found for total consumption of total green vegetables (OR 0.54, 95% CI = 0.34-0.87) and total fruit (OR 0.52, 95% CI = 0.34-0.79) with significant trends in risk. We found that the protective effect of each of these food items was consistently larger among current smokers and among heavy alcohol drinkers, following a multiplicative effect model. In conclusion, this study provides further support to the beneficial effect of high intake of vegetables and fruits on the risk of developing cancers of the oral cavity and oropharynx in Spain, particularly among current smokers and heavy alcohol drinkers. PMID- 12548111 TI - Prevalence of cigarette smoking by birth cohort among males and females in Spain, 1910-1990. AB - We analysed the prevalence of cigarette smoking among subsequent birth cohorts of Spanish males and females. Data were drawn from the Spanish NHIS conducted in 1993 ( = 26 400), 1995 ( = 8300) and 1997 ( = 8300). From the original computer files, the three surveys were pooled to obtain a single sample. A total of 33 223 subjects (16 036 men and 17 187 women) born between 1900 and 1979, >/= 16 years old, were directly interviewed and with complete information on the history of smoking included for analysis. Based on each respondent's sex and calendar year of birth, the person was classified into a particular sex-birth cohort in the decades from 1900-09 to 1970-79. For each year from date of birth to date of survey, respondents were further classified as either cigarette smoker or nonsmoker. The prevalence of cigarette smoking among successive cohorts of Spanish men and women was estimated, with correction for excess mortality of smokers. In men, the peak in smoking was reached in the 1950-59 birth cohort (prevalence rate of 68% at ages 20-29), after increases during the previous calendar years. Smoking among women was rare until 1960. Female smoking prevalence rates increased progressively among subsequent cohorts until 1980. The age distribution of smoking prevalence in women in 1990 mimics that observed in men 40 years earlier. A substantial delay in the spread of the tobacco epidemic among men is apparent. In women, the delay in the initiation of the smoking epidemic ended with a quick diffusion of the habit. PMID- 12548112 TI - Case-control study of female thyroid cancer--menstrual, reproductive and hormonal factors. AB - A case-control study including 204 histologically verified female thyroid cancer patients and an equal number of hospital controls individually matched with cases by sex, age (+/- 2 years), place of residence and time of hospitalization was performed during the period 1996-2000. In the analysis of data, univariate and multivariate conditional logistic regression, methods were applied. According to multivariate analysis, out of hormonal, menstrual and reproductive characteristics, risk factors for thyroid cancer were spontaneous abortions (odds ratio: OR = 1.89, 95% confidence interval (CI) = 1.03-3.50), oral contraceptives use (OR = 2.34, 95% CI = 1.31-4.18) and thyroid enlargement during pregnancy (OR = 16.44, 95% CI = 3.81-70.80). However, none of these three factors remained independently related to thyroid cancer after adjustment for other factors, which were significantly associated with thyroid cancer in the present study (history of residence in endemic goitre area, history of goitre or thyroid nodule, history of other endocrine diseases, radioactive iodine therapy, occupational exposure to various chemicals, family history of thyroid gland diseases and malignant tumours as well as intake of cruciferous vegetables and other vegetables, and consumption of smoked meat and cheese). PMID- 12548113 TI - Consumption of argan oil (Morocco) with its unique profile of fatty acids, tocopherols, squalene, sterols and phenolic compounds should confer valuable cancer chemopreventive effects. AB - The aim of this study was to evaluate the fatty acids, tocopherols, squalene, sterols and phenolic antioxidants in three types of argan oil (Moroccan food, Moroccan aesthetic and a French commercial variety) along with a basic comparison with extra virgin olive and sunflower oil. The fatty acid profiles in the argan oils were very similar, with oleic acid (43%) and linoleic acid (36%) and their respective monoacylglycerols predominating. The major vitamer identified was tocopherol with a mean of 483+/-11 mg/kg, in contrast to -tocopherol, which is the major vitamer in olive (190+/-1 mg/kg) and sunflower oil (532+/-6 mg/kg). The squalene content of the argan oils was very similar with a mean of 313+/-4 mg/100 g, which is lower than that of the olive oil (499 mg/100 g) but significantly higher than in the sunflower oil (6 mg/100 g). In contrast to olive and sunflower oils in which -sitosterol is predominant, the major sterols detected in the argan oils were schottenol (mean 147+/-10 mg/kg) and spinasterol (mean 122+/-10 mg/kg). The only phenolic compounds other than the tocopherol vitamers which could be readily detected and quantitated were vanillic, syringic and ferulic (probably conjugated to glucose) acids along with tyrosol. In contrast to the extra virgin olive oil (793 mg/kg), the concentration of total phenolic compounds is extremely low (<5.0 mg/kg). Nevertheless, argan oil with its high content of the vitamer tocopherol, squalene and oleic acid is likely to enhance the cancer prevention effects of the Moroccan diet. PMID- 12548114 TI - A simulation model for evaluating the medical and economic outcomes of screening strategies for colorectal cancer. AB - Mathematical models have been shown to be useful in predicting the cost effectiveness of cancer screening programmes. We designed a computer macro simulation model aimed at predicting the cost-effectiveness of alternative colorectal cancer screening strategies. This model was built to determine the cost-effectiveness of a biennial screening programme using the Hemoccult test in Burgundy (France). It was validated with data from the Danish randomized study. Estimates of our model showed an extremely close concordance with observed results in the Danish study. The observed mortality reduction was 18.0% and the estimated mortality reduction was 18.4%. Preliminary data from the Burgundy study predict a 14.6% colorectal cancer mortality reduction after 10 years. Sensitivity analyses were performed with different assumptions regarding the participation rates and the lead-time. This model can serve to assess the cost-effectiveness of a variety of screening modalities. PMID- 12548115 TI - Is the spread of HCV in Southern Italy attributable to iatrogenic transmission through unsterile injections? PMID- 12548117 TI - Tolerance induction in rats, using a combination of anti-CD154 and donor splenocytes, given once on the day of transplantation. AB - BACKGROUND: Donor-specific tolerance induction remains an attractive objective that generates much research in the field of transplantation. Unfortunately, most of the protocols available involve pregraft conditioning, making these treatments incompatible with clinical applications. METHODS: LEW.1A rats were grafted with histoincompatible LEW.1W hearts. On the day of transplantation, recipients were treated with anti-CD40L combined with donor splenocytes. The hearts were evaluated for graft survival; cellular infiltrate and intragraft cytokines were determined using real-time reverse transcriptase-polymerase chain reaction. Tolerance induction was assessed by skin grafting and adoptive transfers. RESULTS: The combination of a single injection of anti-CD40L and donor splenocytes, given on the day of surgery, allowed 40% of cardiac allografts to survive long-term (mean survival time=66.3 day). The cellular composition or the extent of graft infiltrate was not modified but was associated with a massive decrease of proinflammatory cytokines expression within the graft. Long-term survivors accepted donor-matched skin grafts, and leukocytes harvested from these animals transferred tolerance into irradiated freshly grafted recipients. CONCLUSION: A combination of costimulation blockade and donor cells, given once at the time of transplantation, is sufficient to induce allograft tolerance in rats. PMID- 12548118 TI - Effects of inhibition of myocardial extracellular-responsive kinase and P38 mitogen-activated protein kinase on mechanical function of rat hearts after prolonged hypothermic ischemia. AB - BACKGROUND: Mitogen-activated protein kinases (MAPKs), including extracellular responsive kinase (ERK) and p38 MAPK, are activated by stresses associated with hypothermia-rewarming and ischemia-reperfusion. Their activation in heart is associated with beneficial (preconditioning) and adverse effects (apoptosis and impaired contractility). This study determined whether ERK and p38 MAPK activities are altered by hypothermic ischemia and normothermic reperfusion and the consequences of their inhibition on recovery of myocardial function. METHODS: Left ventricular work (L x min(-1) x mm Hg) was assessed during normothermic perfusion (30 min) of isolated rat hearts that were either freshly excised or previously subjected to hypothermic storage (8 hr, 3 degrees C) and rewarming (10 min, 37 degrees C) before normothermic reperfusion (30 min). Phospho-specific immunoblot analysis of p38 MAPK was performed in hearts and various cultured cells. RESULTS: Compared with fresh hearts, hearts subjected to hypothermia and rewarming demonstrated impaired left ventricular work (1.96+/-0.53, n=12 vs. 8.37+/-0.46, n=4, <0.05) during reperfusion. The ERK inhibitor, PD98059 (20 microM), present during storage and rewarming, caused modest improvement (3.66+/ 0.75, n=9, <0.05). The p38 MAPK inhibitor, SB202190 (10 microM), when present during reperfusion, improved recovery (to 6.12+/-0.75, n=6, <0.05); it was ineffective if present only during rewarming (1.52+/-0.88, n=4). In rat2 fibroblasts, hypothermia and rewarming activated p38 MAPK and its downstream kinase MAPK-activated protein kinase 2, but not c-Jun N-terminal kinase/stress activated protein kinase. CONCLUSIONS: Myocardial p38 MAPK and MAPK-activated protein kinase 2 are stimulated by hypothermia, ischemia, and rewarming and are detrimental to recovery of mechanical function of hearts subjected to prolonged hypothermic storage. Inhibition of p38 MAPK may be useful in protocols to improve the recovery of mechanical function of cold-stored hearts. PMID- 12548119 TI - Tissue-engineered neomucosa: morphology, enterocyte dynamics, and SGLT1 expression topography. AB - BACKGROUND: The standard therapy for short bowel syndrome is total parenteral nutrition, which is expensive and associated with significant morbidity and mortality. New therapeutic approaches for this disorder are needed. We have applied the techniques of tissue engineering to develop a prototype neointestine. We hypothesized that anastomosis of this neointestine to the native bowel would result in regeneration of mucosal morphology and enterocyte dynamics. METHODS: Biodegradable polymers seeded with neonatal rat intestinal organoid units were implanted into the omenta of adult rats to form neointestinal cysts. Five weeks after implantation, side-to-side cyst-jejunal anastomoses were fashioned in one cohort of rats. Tissues were harvested from all rats at 5 months after implantation. Native jejunal (J) and non-anastomosed (N-N) and anastomosed (A-N) neointestinal tissues were assessed for morphology, epithelial cell proliferation (5-bromo-2-deoxyuridine immunohistochemistry), apoptotic rates (terminal deoxynucleotide transferase-mediated dUTP nick-end labeling assay), and SGLT1 in situ hybridization. RESULTS: Mucosal morphology, rates and topography of enterocyte proliferation, and transporter expression in A-N neointestine recapitulated those of native jejunum. Each of these features was rudimentary in N-N neointestine. CONCLUSIONS: These results suggest that the tissue-engineered neomucosa can develop structural and dynamic features of the normal jejunum. Anastomosis to the native intestine is an essential step for neomucosal development. Tissue engineering offers promise as a novel approach to the treatment of patients suffering from short bowel syndrome. PMID- 12548120 TI - Long-term mycophenolate mofetil monotherapy in combination with calcineurin inhibitors for chronic renal dysfunction after liver transplantation. AB - BACKGROUND: Calcineurin inhibitors (CNIs) are the first-line immunosuppressive agents administered after liver transplantation, but they cause renal impairment. Two recent randomized trials report cellular rejection and liver graft loss when mycophenolate mofetil (MMF) monotherapy was used as a renal-sparing agent. Our experience with MMF in the same setting but with longer follow-up is described. METHODS: In 45 patients with serum creatinine more than 120 micromol/L or creatinine clearance less than 50 mL/min, 2 g MMF per day was administered (median 29 months, 1-49 months) either as monotherapy (with all other immunosuppression withdrawn in 1 month) in 16 patients (group I) or in combination with low-dose CNI (trough tacrolimus 100 days) and regulatory cells were generated. In vitro, AT-III suppressed proliferation of mixed leukocyte responses and generation of interleukin-2. CONCLUSION: AT-III can be not only an antithrombotic agent but also a strong immunomodulating agent when used at high dose. PMID- 12548127 TI - Origin and subset distribution of peripheral blood dendritic cells in patients with chronic graft-versus-host disease. AB - BACKGROUND: After allogeneic hematopoietic stem cell transplantation, donor T cells interact with an antigen-presenting cell environment that is distorted in number, level of activation, and origin. The role of antigen presentation in the development of chronic graft-versus host disease (cGVHD) is unknown. METHODS: The number and origin of peripheral blood immature myeloid (CD19- CD1c+) and plasmacytoid (BDCA-2+) dendritic cells (DCs) was determined in 30 patients at more than 100 days after allogeneic hematopoietic stem cell transplantation. RESULTS: Patients with cGVHD had significantly higher plasmacytoid DC numbers than individuals without this complication (9.1+/-2.0 x 10(6)/L versus 3.8+/-0.6 x 10(6)/L, =0.025). Chimerism studies demonstrated that DCs in patients with cGVHD were exclusively of donor origin, whereas persistence of host DCs was observed in some control patients. CONCLUSIONS: The antigen-presenting cell environment in patients with cGVHD, as represented by immature blood DCs, is of donor origin but distorted in terms of subset distribution. PMID- 12548128 TI - MALToma: a Helicobacter pylori-associated malignancy in transplant patients: a report from the Israel Penn International Transplant Tumor Registry with a review of published literature. AB - BACKGROUND: Mucosa-associated lymphoid tissue lymphoma (MALToma) is a Helicobacter pylori-related tumor of B-cell origin, the malignant potential for which remains to be defined in immunosuppressed patients. METHODS: Review of the Israel Penn International Transplant Tumor Registry identified six cases of gastric MALToma. Patient demographics, management, and outcomes were compared and published literature was reviewed. RESULTS: MALToma developed in six transplant recipients (three kidney, two heart, one kidney-pancreas). All were treated with immunosuppression minimization and therapy for H. pylori, resulting in disease regression in five patients. One patient developed progression to high-grade MALToma despite documented H. pylori eradication, required surgery and chemotherapy, and died, with significant disease at autopsy. CONCLUSIONS: Treatment of MALToma with immunosuppression minimization and anti-H. pylori therapy results in a majority of patients becoming disease free. Observation of malignant degeneration into an aggressive, high-grade lymphoma in one patient indicates the malignant potential. Diligent follow-up of these patients with endoscopy and biopsy is therefore indicated. PMID- 12548129 TI - Randomized controlled trial of oral ganciclovir versus oral acyclovir after induction with intravenous ganciclovir for long-term prophylaxis of cytomegalovirus disease in cytomegalovirus-seropositive liver transplant recipients. AB - BACKGROUND: Without effective antiviral prophylaxis, cytomegalovirus (CMV) disease is a common cause of morbidity and mortality after liver transplantation. In this randomized, controlled trial, we compared the efficacy and safety of oral ganciclovir with oral acyclovir after induction with intravenous (IV) ganciclovir for long-term prophylaxis of CMV disease in CMV-seropositive liver transplant recipients. METHODS: Patients were initially administered IV ganciclovir at a dose of 6 mg/kg per day from day 1 to day 14 after transplantation followed by either oral ganciclovir (1 g every 8 hr) or oral acyclovir (800 mg every 6 hr) from day 15 to day 100 after transplantation. RESULTS: CMV disease occurred in only 1 of 110 patients (0.9%) receiving ganciclovir compared with 8 of 109 patients (7.3%) receiving acyclovir within the first year after transplantation (P =0.019). There was one case of CMV colitis in the ganciclovir group, whereas four cases of CMV syndrome, three cases of CMV pneumonia, and one case of CMV hepatitis developed in the acyclovir group. The only death from CMV disease occurred in an acyclovir-treated patient with CMV pneumonia. Both oral ganciclovir and oral acyclovir were generally well tolerated. Reversible leukopenia (decline in white blood cell count to <3.0 x 10(9)/L) was more common with oral ganciclovir (38/110 patients, 35%) than with oral acyclovir (20/109 patients, 18%) (P =0.009). The emergence of ganciclovir-resistant strains of CMV was not found during the study. CONCLUSIONS: A prophylactic regimen of 2 weeks of IV ganciclovir followed by an additional 12 weeks of oral ganciclovir is superior to a similar regimen of IV ganciclovir followed by oral acyclovir and almost completely eliminates CMV disease after liver transplantation. This superior protection against CMV disease extends up to 1 year after transplantation and is not associated with ganciclovir resistance. PMID- 12548130 TI - Successful surgical salvage of pancreas allograft. AB - BACKGROUND: Early and late complications related to the pancreas after simultaneous kidney-pancreas transplantation (SKPT) frequently result in graft loss. The authors describe a surgical rescue technique that allows salvage of the pancreatic graft when surgical complications appear after the transplant. METHODS: Of 158 patients who underwent SKPT, 7 were identified with posttransplant complications that required surgical salvage of the pancreas allograft. The surgical salvage technique consisted of the following: pancreatoduodenectomy with conversion from whole-pancreas transplant with bladder or enteric diversion to segmental graft with duct injection (three cases) and conversion from whole-pancreas transplant with duct injection (four cases). RESULTS: Five of seven pancreas allografts are still functioning, with a mean follow-up of 28 months (range, 6-42 months). CONCLUSION: The described surgical treatment may be useful for surgical salvage of the pancreatic allograft, without major impairment of endocrine function. PMID- 12548131 TI - Diagnosis and treatment of prostate cancers in renal-transplant recipients. AB - BACKGROUND: There is no consensus regarding prostate cancer in renal-transplant recipients (RTR). A questionnaire evaluating prostate cancer screening after transplantation and assessing the number, diagnostic modalities, treatment, and outcome of prostate cancer cases was mailed to 22 French renal-transplant centers. RESULTS: Among 1,680 RTR in 1998, 11 (0.65%) cases of prostate cancer were diagnosed, and among the 2,338 recipients followed up, 28 (1%) cases of prostate cancer have been diagnosed and treated. Median ages at diagnosis and at transplantation were 63 and 58, respectively. Clinical stages were T1 50% and T2 25%. Eighteen patients had a Gleason score under 7. At 18 months of mean follow up, 2 men had died from prostate cancer, and in the curative treatment group, 16 of 17 men were alive with no evidence of disease. CONCLUSIONS: The incidence of prostate cancer in RTR appeared to be higher than expected. Prostate specific antigen (PSA) testing should be performed routinely each year in renal transplantation centers. PMID- 12548133 TI - Intravenous immunoglobulin-induced panel reactive antibody A reduction: not all preparations are created equal. AB - BACKGROUND: Use of intravenous (IV) immunoglobulin (Ig) to obtain panel reactive antibody (PRA) A reduction in sensitized patients has been widely reported. Because no IVIg preparation is formulated specifically for this purpose, the authors have sought to determine whether, through laboratory testing, they could guide the rational choice of product for clinical use. METHODS: Using a flow cytometric approach, the authors have quantitatively determined the capacity of 22 different IVIg preparations to cause PRA reduction. RESULTS: IVIg preparations showed considerable variability in their individual capacity to reduce serum PRA. Protein-A pretreatment of IVIg preparations was found to reduce their capacity to cause PRA reduction. CONCLUSION: Laboratory screening of IVIg preparations provides a rational basis for the selection of product for administration to patients in whom the aim is to produce a PRA reduction. Experiments involving protein-A treatment of IVIg preparations indicate that immunoglobulin G is the principal factor involved in the abrogation of serum reactivity. PMID- 12548132 TI - Blockade of in vivo VEGF-KDR/flk-1 signaling does not affect revascularization of freely transplanted pancreatic islets. AB - BACKGROUND: The aim of this study was to analyze the role of vascular endothelial growth factor (VEGF) binding to its high-affinity receptor kinase insert domain (KDR)-containing receptor/fetal liver kinase (flk)-1 in mediating revascularization of freely transplanted pancreatic islets in vivo. METHODS: Isolated pancreatic islets were syngeneically transplanted into dorsal skinfold chambers of Syrian hamsters. Animals were treated daily with the VEGF-KDR/flk-1 antagonist SU5416 (25 mg/kg intraperitoneally) or received vehicle for control. Intravital fluorescence microscopy and computer-assisted off-line analysis were used to study islet graft angiogenesis and revascularization during days 6, 10, and 14 after transplantation. RESULTS: In controls, islets were revascularized within 10 to 14 days after transplantation. SU5416 treatment did not affect revascularization, inasmuch as both functional capillary density and size of revascularized endocrine tissue did not differ from that of vehicle-treated controls. CONCLUSION: Because blockade of VEGF-KDR/flk-1 function does not affect islet revascularization, we conclude that VEGF signaling through its high affinity receptor KDR/flk-1 is not an essential prerequisite for the process of new-vessel formation in freely transplanted islets of Langerhans. PMID- 12548134 TI - Posttransplant epididymal lymphoma: an aggressive variant. PMID- 12548135 TI - Modified liver hanging maneuver during orthotopic liver transplantation with inferior vena cava preservation. PMID- 12548136 TI - The impact of body mass index on renal transplant outcomes: a significant independent risk factor for graft failure and patient death. PMID- 12548137 TI - Nonaltruistic kidney donations in contemporary Jewish law and ethics. AB - BACKGROUND: In 2000, the Consensus Statement on the Live Organ Donor reported that "direct financial compensation for an organ from a living donor remains controversial and illegal in the United States" and took note of the position of the Transplantation Society that "Organs and tissue should be given without commercial consideration or commercial profit." Christian authorities insist that organ donors must not accrue economic advantage, and "selling" organs deprives the donation of its ethical quality. METHODS: The writings of major contemporary authorities of Jewish law and ethics whose halakhic positions on bioethical issues are regularly considered by Orthodox, Conservative, and Reform ethicists were reviewed. Their positions on this issue were contrasted with those of various contemporary secular and religious authorities. RESULTS: These Jewish authorities reject the notion that generosity and charity, rather than monetary gain and greed, must serve as the exclusive basis for donation of functioning organs. CONCLUSION: Although nonaltruistic sale of kidneys may be theoretically ethical, ultimately its ethical status in Jewish ethics and law is inextricably connected to solving a series of pragmatic programs, such as creating a system that ensures that potential vendors and donors are properly informed and not exploited. Lacking such arrangements, ethical nonaltruistic kidney donations remain but a theoretical possibility. PMID- 12548138 TI - Acyl glucuronide drug metabolites: toxicological and analytical implications. AB - Although glucuronidation is generally considered a detoxification route of drug metabolism, the chemical reactivity of acyl glucuronides has been linked with the toxic properties of drugs that contain carboxylic acid moieties. It is now well documented that such metabolites can reach appreciable concentrations in blood. Furthermore, they are labile, undergo hydrolysis and pH-dependent intramolecular acyl migration to isomeric conjugates of glucuronic acid, and may react irreversibly with plasma proteins, tissue proteins, and with nucleic acids. This stable binding causes chemical alterations that are thought to contribute to drug toxicity either through changes in the functional properties of the modified molecules or through antigen formation with subsequent hypersensitivity and other immune reactions. Whereas in vitro data on the toxicity of acyl glucuronides have steadily accumulated, direct evidence for their toxicity in vivo is scarce. Acyl glucuronides display limited stability, which is dependent on pH, temperature, nature of the aglycon, and so on. Therefore, careful sample collection, handling, and storage procedures are critical to ensure generation of reliable pharmacologic and toxicologic data during clinical studies. Acyl glucuronides can be directly quantified in biologic specimens using chromatographic procedures. Their adducts with plasma or cell proteins can be determined after electrophoretic separation, followed by blotting. ELISA techniques have been used to assess the presence of antibodies against acyl glucuronide-protein adducts. This review summarizes the most recent evidence concerning biologic and toxicologic effects of acyl glucuronide metabolites of various drugs and discusses their relevance for drug monitoring. A critical evaluation of the available methodology is included. PMID- 12548139 TI - Utility of antiepileptic drug monitoring in the pediatric emergency department. AB - Convulsive disorders are common in the pediatric age group, and measurement of serum concentration of an antiepileptic drug (AED) is frequently ordered for epileptic patients in the emergency department (ED). The objective of this study was to develop a better understanding of the indications for, and consequences of, monitoring AED serum concentrations in the pediatric ED. Charts of 116 patients who visited the ED and were tested for blood levels of AED were retrospectively reviewed. Main outcome measures were number and percentage of levels outside the therapeutic range, discontinuation of an AED or introduction of a new one, dosage modifications, and admission to hospital. Two pediatricians and a pediatric neurologist aware only of patients' age, weight, diagnosis, history, clinical presentation, and drug details reviewed each case and on the basis of predetermined criteria decided whether measurement of AED was indicated. Mean age (+/- SD) of the study population was 7 +/- 5 years (range, 2 months-17 years). Forty-two patients (36%) were on monotherapy, and 74 (64%) were on polytherapy. Sixty-eight patients (59%) presented with increased seizure frequency, 7 (6%) with status epilepticus, and 13 (11%) with suspected AED toxicity. The remainder of the children presented with problems unrelated to epilepsy. No significant difference was found between patients with AED levels within the therapeutic range and those with levels outside it in the proportion of children needing dosage change, change in medication, or hospital admission (P = 0.5, 0.8, and 0.8, respectively). None of the patients presenting with status epilepticus and only 15% of those with increased seizure activity had subtherapeutic levels. Review of the cases suggested that measuring serum AED level was not indicated in 57 (49.1%) patients. In the pediatric ED, abnormal AED levels do not correlate with clinical management. Before ordering tests, physicians should consider whether their results would alter patient treatment. PMID- 12548140 TI - Quantification of immunosuppression by flow cytometry in stable renal transplant recipients. AB - The current standard of monitoring transplant patients by drug levels is not optimal because it does not take into account the different and individual effects of immunosuppressive drugs on each patient. In this study, the authors tested immune function assays for monitoring transplant patients. Blood was collected from stable renal transplant patients treated with cyclosporin, mycophenolate mofetil, and prednisone (n = 8), and from healthy volunteers (n = 12). Lymphocyte proliferation, expression of T-cell surface activation antigens (CD25, CD71, CD11a, CD95, CD154), production of intracellular cytokines (IL-2, INFgamma, TNFalpha), and lymphocyte subsets (CD4, CD8, CD16, CD20) were assessed by flow cytometry. Lymphocyte proliferation, expression of T-cell surface activation antigens, and production of intracellular cytokines were significantly decreased in transplant recipients compared with healthy control volunteers. The combined effects of several immunosuppressive drugs in renal transplant recipients can be quantitated with immune function assays in whole blood. This new method may be helpful to achieve an optimal level of immunosuppression for each patient. PMID- 12548141 TI - Bayesian forecasting of oral cyclosporin pharmacokinetics in stable lung transplant recipients with and without cystic fibrosis. AB - The aims of the current study were (1) to study Neoral pharmacokinetics (PK) in stable lung recipients with or without cystic fibrosis (CF), (2) to compare Neoral PK between these two groups, and (3) to design Bayesian estimators for PK forecasting and dose adjustment in these patients using a limited number of blood samples. The individual PK of 19 adult lung transplant recipients, 9 subjects with CF and 10 subjects without CF, were retrospectively studied. Three profiles obtained within 5 days were available for each patient. A PK model combining a gamma distribution to describe the absorption profile and a two-compartment model were applied. Different exposure indices were estimated using nonlinear regression and Bayesian estimation. The PK model developed reliably described the individual PK of Neoral in lung transplant patients with and without CF, and the values of the first and second half-lives were different in these two populations (lambda(1) = 4.14 +/- 3.01 vs. 2.16 +/- 1.75 h(-1); P < 0.01; lambda(2) = 0.36 +/ 0.11 vs. 0.49 +/- 0.12 h(-1); P < 0.01), while the mean absorption time and standard deviation of absorption time tended to be less in patients with cystic fibrosis (P < 0.1). Also, the patients with CF required higher doses than those without CF to achieve similar drug exposure. Consequently, population modeling was performed in CF and non-CF patients separately. Bayesian estimation allowed accurate prediction of AUC(0-12), AUC(0-4), C(max), and T(max) using three blood samples collected at T0h, T1h, and T3h in both groups. This study demonstrated the applicability and good performance of the PK model previously developed for oral cyclosporin and of the MAP Bayesian estimation of cyclosporin systemic exposure in CF and non-CF patients. Moreover, it is the first to propose a monitoring tool specifically designed for cyclosporin monitoring in patients with CF. PMID- 12548142 TI - A Bayesian method based on clotting factor activity for the prediction of maintenance warfarin dosage regimens. AB - A Bayesian algorithm, employing a population pharmacokinetic-pharmacodynamic model, for the effective and rapid prediction of warfarin maintenance dosing requirements was developed. The algorithm was evaluated prospectively in five healthy volunteers who were given a 15-mg single dose of racemic warfarin. Based on previous population pharmacokinetic and pharmacodynamic parameters and factor VII response measurements taken during the first 48 hours, dosage regimens to achieve a subtherapeutic degree of anticoagulation (50% of normal) were determined. Three factor VII response measurements were sufficient to determine dosing requirements in the five volunteers. The advantage of the algorithm is that it does not require warfarin concentration measurements and uses non-steady state data. PMID- 12548143 TI - Cross-reactivity assessment of carbamazepine-10,11-epoxide, oxcarbazepine, and 10 hydroxy-carbazepine in two automated carbamazepine immunoassays: PETINIA and EMIT 2000. AB - This study was conducted to compare the cross-reactivity of two commercially available carbamazepine (CBZ) immunoassays (PETINIA and EMIT 2000) with carbamazepine-10,11-epoxide (CBZ-E), the active metabolite of CBZ. Oxcarbazepine (OCBZ) and its main metabolite 10-hydroxy-carbazepine (HCBZ) have a chemical structure closely related to that of CBZ. The cross-reactivities of these two drugs were also investigated. In the first part of the study, Lyphocheck blank human serum and Chemonitor quality controls (containing CBZ without CBZ-E) were spiked with variable amounts of CBZ-E. The apparent CBZ levels were measured by PETINIA and EMIT 2000 methods. The interference from OCBZ and HCBZ was directly assessed by measuring the apparent CBZ levels in Chromsystems Trileptal quality controls (containing OCBZ and HCBZ). In the second part of the study, the CBZ levels of serum samples from 49 patients, including 2 patients with massive CBZ ingestion, were measured by immunoassays and compared with a high-pressure liquid chromatography (HPLC) reference technique allowing the simultaneous measurement of CBZ and CBZ-E. The antibody used in the PETINIA assay cross-reacts (about 90%) with CBZ-E. In one case of CBZ poisoning (CBZ and CBZ-E levels measured by HPLC were 26.2 and 18.2 mg/L, respectively), CBZ level measured by PETINIA was falsely elevated (42.5 mg/L). In contrast, the specificity of EMIT 2000 was satisfactory (29.5 mg/L). The two immunoassays tested showed low cross-reactivity with OCBZ and HCBZ. In conclusion, it appears that the CBZ-E metabolite present in samples can falsely increase CBZ levels measured by the PETINIA assay. PMID- 12548144 TI - Therapeutic drug monitoring of olanzapine: the combined effect of age, gender, smoking, and comedication. AB - Therapeutic drug monitoring (TDM) data for the antipsychotic drug olanzapine were investigated with respect to concentration versus dose relationship, intraindividual versus interindividual variability, and the combined influence of patient characteristics on steady-state concentration. The study included 250 patients, with daily doses ranging from 2.5 to 30 mg. Median concentration to dose ratio was 2.1 (ng/mL)/(mg/d), with 90% of the distribution in a fivefold range. In the first subgroup of patients with two measurements at different doses (n = 21), data were in keeping with linear concentration versus dose relationship. In the second subgroup of patients with repeated measurements at a constant daily dose (n = 40), estimates of within-patient and between-patient variabilities were 30.5% and 49.4%, respectively. In the whole sample, multiple regression analysis of dose-normalized concentration revealed significant effects of time postdose (-18% per 12 hours delay, P < 0.05), age >/=60 years (+27%, P < 0.005), cigarette smoking (-12%, P < 0.05), and comedication with fluvoxamine (+74%, P < 0.001), paroxetine, fluoxetine, or sertraline (considered together, +32%, P < 0.05), venlafaxine (+27%, P < 0.05), and inducers of P450 enzymes ( 40%, P < 0.001). The final model included a tendency for higher concentration associated with female gender (+11%, P = 0.07) and accounted for 27% of observed interindividual variability. When considering a worst-case scenario, an elderly, nonsmoking woman prescribed fluvoxamine comedication was predicted to reach a 4.6 fold higher olanzapine concentration than a young male smoker coadministered carbamazepine. The current study suggests that patients characterized by a combination of factors associated with altered metabolism may benefit from olanzapine TDM. PMID- 12548145 TI - Plasma gabapentin concentrations in children with epilepsy: influence of age, relationship with dosage, and preliminary observations on correlation with clinical response. AB - The influence of age and administered daily dosage on the plasma concentrations of gabapentin (GBP) at steady state was evaluated in a group of 41 children and young adults (aged 3-30 years) receiving long-term adjunctive treatment with GBP for the management of refractory partial-onset seizures. For each patient, peak and trough concentrations were determined by a specific high-performance liquid chromatography (HPLC) method in samples obtained before the morning dose and 2.5 hours later, respectively. To assess within-subject relationship between plasma concentration and dosage, 30 patients were evaluated at more than one dosage level. Within the assessed dose range, plasma GBP concentrations were linearly related to dose. Apparent oral clearance values (mean +/- SD) in children aged 6 years or less (4.8 +/- 0.9 mL/kg/min) were comparable with those observed in children aged 7 to 15 years (4.6 + 1.5 mL/kg/min) and moderately higher than those found in young adults (3.9 + 0.9 mL/kg/min), even though differences among groups failed to reach statistical significance. There was, however, a significant difference in CL/F between children aged 10 years or less and older children (5.1 +/- 1.1 vs. 3.8 +/- 1.2 mL/kg/min, P < 0.005). Of the 41 patients who entered the study, 22 discontinued treatment, mostly due to insufficient efficacy. No significant difference in plasma GBP concentration was detected between patients showing a greater than 50% reduction in seizure frequency (4.1 +/- 1.9 microg/mL, n = 11, mean +/- SD) and those having no significant clinical improvement (4.4 +/- 1.7 microg/mL, n = 30). These results indicate that in children given dosages up to 50 mg/kg/d (mean, 25 mg/kg/d), GBP pharmacokinetic analyses show no important deviation from linearity. The data also suggest that, on average, children may need moderately higher dosages to reach plasma GBP concentrations comparable with those found in adults. There seems to be a large variation in the plasma concentrations of the drug associated with a favorable therapeutic response. PMID- 12548146 TI - Correlation of levetiracetam concentrations between serum and saliva. AB - PURPOSE: More than 100 drugs have been evaluated for salivary therapeutic drug monitoring since the 1970s. The most studied drugs are the anticonvulsants phenytoin, phenobarbital, and carbamazepine, demonstrating strong correlations between serum and saliva concentrations. No published data exist for levetiracetam (LEV) to the authors' knowledge. This study's objective is to determine the correlation between LEV serum and saliva concentrations. METHODS: Investigators identified subjects seen in neurology clinics at the University of Kentucky. Patients were eligible if they agreed to participate in this study, were taking LEV for a minimum of 4 weeks, and if a serum LEV concentration had been ordered by their physician. Patients spit a minimum of 0.25 mL into a cup to obtain saliva samples. Blood samples were obtained by phlebotomy. RESULTS: Serum and saliva LEV concentrations were determined via high-performance liquid chromatography (HPLC) in two separate reference laboratories. Linear regression analysis was used to evaluate correlations. Serum and saliva samples were obtained from 40 patients (22 female, 18 male), ranging from 3 to 57 years of age. The mean +/- SD serum LEV concentration for reference laboratory A was significantly lower (P < 0.0001) than reference laboratory B, 23.6 +/- 13.8 microg/mL and 27.0 +/- 16.9 microg/mL, respectively. The mean +/- SD saliva to serum concentration fraction was also different for the two laboratories, i.e., 41.0% +/- 0.15% for lab A and 36.0% +/- 0.15% for lab B (P = 0.001). The correlation coefficients for the two laboratories were similar, 0.87 and 0.86 (both P < 0.0001) for labs A and B, respectively. CONCLUSION: A significant positive correlation exists between LEV saliva and serum concentrations. The ability to monitor LEV therapy using saliva may provide benefits that include facilitating sample collection and improving the quality of life for persons with epilepsy. Patients with poor venous access, such as children and elderly patients, and persons afraid of needles may particularly benefit from this method. PMID- 12548147 TI - Evaluation of estimated and measured creatinine clearances for predicting the pharmacokinetics of vancomycin in adult liver transplant recipients. AB - This study examined the pharmacokinetics and dosing requirements of vancomycin in adult liver transplant recipients and also evaluated the predictability of determining vancomycin-dosing requirements utilizing an estimated creatinine clearance (CrCl) approach. Twenty adult liver transplant recipients were included in this analysis. Vancomycin pharmacokinetic parameters and dosing requirements calculated from estimated CrCl and population-based pharmacokinetic equations were compared with values calculated using serum concentrations and assuming a one-compartment model. Compared with the values obtained using equations to estimate the CrCl and vancomycin pharmacokinetics (t, Cl, and Vd), the actual values were statistically different for half-life and clearance (11.0 vs. 16.4 hours and 52 vs. 36 mL/min, respectively; P < 0.01). Additionally, CrCl that were estimated using population-based formulas significantly overestimated actual CrCl calculated using 24-hour urine collections (65-78 vs. 43 mL/min; P < 0.05). The results from this study indicate that serum creatinine concentrations do not adequately predict glomerular filtration rates (GFR) or vancomycin clearance in adult liver transplant recipients. Based on these results, the use of 24-hour urine CrCl to predict GFR and serum concentrations to properly dose vancomycin is advocated. PMID- 12548148 TI - Treatment failure of nelfinavir-containing triple therapy can largely be explained by low nelfinavir plasma concentrations. AB - The relationship between plasma concentrations of nelfinavir and virologic treatment failure was investigated to determine the minimum effective concentration of nelfinavir. Plasma samples were prospectively collected from treatment-naive patients who began taking nelfinavir, 1,250 mg BID + two nucleoside reverse transcription inhibitors (NRTIs). Nelfinavir concentration ratios were calculated by dividing each individual nelfinavir level by the time adjusted population value. Virologic failure was defined as either no response (a detectable viral load after 6 months) or a relapse (detectable viral load after being undetectable, or an increase in viral load >1 log above nadir). Forty-eight patients were included with a median follow-up period of 8 months. The median concentration ratio of nelfinavir was 0.98 (interquartile range, 0.76-1.47). Virologic failure was observed in 29% of the patients. In a univariate analysis, the nelfinavir concentration ratio appeared to be the single determinant that was related to virologic failure (P = 0.039). Patients with a median ratio <0.90 had a relative risk of 3.0 (95% CI, 1.2-7.6) for virologic failure. Using this threshold, virologic failures were detected with 64% sensitivity and 74% specificity (P = 0.014). Virologic failure of nelfinavir-containing triple therapy can be explained, to a large extent, by low plasma levels of nelfinavir. PMID- 12548149 TI - In vitro stability study of methotrexate in blood and plasma samples for routine monitoring. AB - A study was conducted to assess the stability of methotrexate (MTX) in pooled plasma and whole blood samples during an 11-day period. The concentrations were determined by liquid chromatography with UV detection over time, in the dark or in light, at different temperatures (4 degrees C or room temperature), and for a low and a high concentration level (0.6 and 2.5 microg/mL, 1.32 and 5.5 micromol/L, respectively). The accuracy, linearity, and variability of the method were assessed. This work demonstrates that blood samples are stable for 2 days at room temperature and for 6 days at 4 degrees C. Furthermore, there is no significant loss (<17%) in plasma at any of the investigated temperatures for at least 6 days. This study shows that whole blood samples can be stored for 3 days before centrifugation and that plasma samples can be reanalyzed after 6 days without any loss. PMID- 12548150 TI - Sensitive assay for the determination of fluvastatin in plasma utilizing high performance liquid chromatography with fluorescence detection. AB - The authors describe a rapid, useful, specific, and very sensitive high performance liquid chromatographic assay for the determination of fluvastatin (FV) level using atorvastatin as the internal standard (IS). After a simple deproteinization of 1.0 mL of plasma with acetonitrile, the drug and IS were extracted with tert-methyl butyl ether (TMBE). An efficient separation was performed using an 8 mm x 10 cm Nova Pak C(18) 4-microm particle size radial compression cartridge. The mobile phase consisted of an aqueous solution containing 20 mmol/L dibasic sodium dihydrogen phosphate with 1 mmol/L sodium lauryl sulfate adjusted to pH 7 with phosphoric acid and acetonitrile (70:30 v/v) delivered at a flow rate of 1.0 mL/min. The compounds of interest were detected using a fluorescence detector with the excitation wavelength set at 305 nm and the emission at 380 nm. Under these conditions, the retention times for FV and IS were 8.8 and 10.6 minutes, respectively. The concentration of FV in plasma was linear (r > 0.999) for the wide range that was examined (0.5-1,000 ng/mL). The recovery ranged from 88% to 96%. This sensitive, rapid, and simple analytical method gives accurate results over the wide range of concentrations examined. This method is used currently for clinical therapeutic monitoring and pharmacokinetic studies of FV in patients with hypercholesterolemia. PMID- 12548151 TI - Pharmacokinetics of a single oral dose of baclofen in pediatric patients with gastroesophageal reflux disease. AB - Transient relaxation of the lower esophageal sphincter (TLESR) is the predominant mechanism of gastroesophageal reflux (GER) in adults and children. Baclofen [4 amino-3-(p-chlorophenyl)-butanoic acid], a gamma-aminobutyric acid (GABA)-B receptor agonist used for the management of spasticity, has been recently shown to significantly inhibit GER in healthy adults without any relevant side effects. The objective of this study was to evaluate the pharmacokinetics of baclofen in a pediatric population with GER disease. In an open-label single-dose pharmacokinetic study, eight children with the diagnosis of GER made on clinical grounds received an oral dose of baclofen, 2.5 mg. Blood samples were drawn from an indwelling venous catheter, and urine was collected during a postdose period of 8 hours. The concentration of baclofen in these body fluids was determined using a validated high-performance liquid chromatography (HPLC) method with electrochemical detection after OPA-sulfite derivatization. Pharmacokinetic data were analyzed using the nonlinear regression program Scientist. Serum concentration-time curves could be best described using a two-compartment open model with a lag time. Mean plasma clearance (Cl) was 315.9 mL/h/kg; volume of distribution (Vd) was 2.58 L/kg; and half-life (T(1/2)beta) was 5.10 hours. No side effects were noted. As half-lives were comparable with those found in adult studies, the risk for accumulation seems not greater in children than in adults. Body composition can have a strong influence on the Vd of baclofen and, therefore, on the dose needed to obtain therapeutic plasma levels. Dosing according to clearly defined age groups with the help of therapeutic drug monitoring seems preferable. In view of the negative correlation between body weight and Vd, dosing according to body weight using adult pharmacokinetic data does not seem an effective way for using baclofen in children. PMID- 12548152 TI - Use of high-performance liquid chromatography with diode-array detection after a primary drug screening in patients admitted to the emergency department. AB - Two hundred fifty samples of patients admitted to the Emergency Department at the Hospital Municipal de Urgencias, Cordoba, Argentina for a drug screening by HPTLC, FPIA, spectrophotometric methods, and HPLC/DAD were randomly selected. The rate of positive screens was 34.0% with the following rate distribution: 12.0% alcohol, 13.6% nonsteroid antiinflammatory drugs (NSAIDs), 2.0% anticonvulsants, 0.8% barbiturates, 0.4% narcotics, 0.4% antidepressants, 2.8% cocaine, and 2.0% cannabinoids. Psychoactive drugs (alcohol, cocaine, and cannabinoids) were detected in 43.9% of the patients admitted for traffic accidents, namely 35.71% alcohol, 2.38% alcohol-cocaine, 2.38% alcohol-cannabinoids, 2.38% cocaine cannabinoids, and 2.38% alcohol-cocaine-cannabinoids. These results help identify the trend of use and/or abuse of drugs and its relationship with different causes of admission (accidents, overdose, and other pathologies), age, and gender. PMID- 12548153 TI - Frequency distribution of C3435T mutation in exon 26 of the MDR1 gene in a Spanish population. AB - P-glycoprotein (PGP) is a transmembrane efflux transporter with an important role in drug therapy. The level of PGP expression leads to relevant consequences in terms of efficacy and toxicity by modulating drug disposition. A single nucleotide polymorphism (SNP) in exon 26 of the gene C3435T was recently associated to PGP levels and substrate uptake. Persons who were homozygous for the T-allele had significantly decreased PGP expression compared with C/C persons. Due to this fact and bearing in mind the important differences among populations regarding the frequencies of persons carrying mutations affecting drug disposition, the authors wanted to study the prevalence of this genetic trait in their population. DNA samples from 408 persons were assayed by a PCR RFLP method. The results showed that the distributions of the C/C, T/T, and C/T genotypes in the Spanish population were 26%, 22%, and 52%, respectively. With regard the C-allele frequency, which has been studied in several populations, the result in their population was 52%, significantly lower (P < 0.0001) than that found in African populations and similar to several Asian and Caucasian (UK) populations (P > 0.05). By contrast, the C-allele frequency in southwest Asian, German, and Portuguese populations was significantly lower than in the Spanish population (P < 0.001, P < 0.01, and P < 0.05, respectively). The great differences found between their population and others, such as the African and southwest Asian populations, could have important therapeutic implications when drugs that are a substrate of PGP are used. PMID- 12548154 TI - Lack of clinically significant interference by spironolactone with the AxSym Digoxin II assay. PMID- 12548155 TI - Cesium toxicity: a case of self-treatment by alternate therapy gone awry. AB - Cesium salts have been used in animal models to induce cardiac arrhythmias for several decades, but the sequelae of human cesium toxicity have seldom been described. The authors describe a case of cesium toxicity manifested by syncope, polymorphic ventricular tachycardia, hypokalemia, and a QT interval prolonged to 650 milliseconds that resolved over 4 days following withdrawal of cesium. The patient had a 2-year history of colon cancer and had self-treated with cesium chloride, 3 g/d, for several weeks, using cesium as a form of alternate therapy for cancer. The authors describe the pathophysiologic correlates and risks of cesium consumption and conclude that cesium toxicity should be considered among the differential diagnoses of prolonged QT interval. PMID- 12548156 TI - Phenytoin decreases the blood concentrations of sirolimus in a liver transplant recipient: a case report. AB - This report documents that coadministration of phenytoin leads to decreased blood concentrations and area under the blood concentration-time curve of sirolimus in a liver transplant patient. It is essential to monitor the blood concentrations of sirolimus and adjust the sirolimus dosage when phenytoin administration begins or ends. PMID- 12548157 TI - High-performance liquid chromatographic analysis of the anticancer drug irinotecan (CPT-11) and its active metabolite SN-38 in human plasma. AB - A rapid, sensitive, and specific high-performance liquid chromatography (HPLC) method for the simultaneous determination of irinotecan (CPT-11) and its active metabolite SN-38 in human plasma is described. The analytes are quantified as the totals of their carboxylate and lactone form. The sample pretreatment consisted of a simple protein precipitation with acetonitrile-methanol (1:1, v/v), after which CPT-11 and SN-38 were quantitatively converted to their carboxylate form by adding 0.01 mol/L sodium tetraborate (pH, 9). Chromatography was carried out on a Zorbax SB-C18 column with fluorescence detection. The method has been validated, and stability tests under various clinically relevant conditions have been performed. The lower limit of quantification (LLOQ) was 5.0 ng/mL for CPT-11 and 0.5 ng/mL for SN-38. Standard concentration ranges were linear between 5 and 1,500 ng/mL for CPT-11 and between 0.5 and 100 ng/mL for SN-38. This assay is simple, rapid, and very useful for therapeutic monitoring of CPT-11 and SN-38. PMID- 12548158 TI - A preliminary study of the effect of fluoxetine treatment on the 2:16-alpha hydroxyestrone ratio in young women. AB - Cytochrome P450 isoenzymes are known to contribute to estrone metabolism. The authors hypothesized that fluoxetine, a known inhibitor of multiple P450 isoenzymes including 3A4, 2C9, and 2D6, would affect estrone metabolism, altering the 2-hydroxyestrone:16-alpha-hydroxyestrone (2OHE1:16OHE1) ratio. In this preliminary study, four of eight recruited women with regular menstrual cycles, aged 21-37 years, completed a 24-hour urine collection prior to initiation of fluoxetine therapy and after at least 5 weeks of antidepressant treatment. In three of the four women who were nonsmokers, the 2OHE1:16OHE1 ratio was significantly higher after 5 weeks of fluoxetine therapy (pretreatment, 2.08 +/- 0.11; posttreatment, 3.50 +/- 0.46; paired Student's t-test P = 4.72, P = 0.021). PMID- 12548159 TI - CDX-2 homeobox gene expression is a reliable marker of colorectal adenocarcinoma metastases to the lungs. AB - Lung metastases from colorectal carcinomas (CRC) can be resected with improved survival. The distinction between primary lung adenocarcinomas and metastases from CRC may sometimes be difficult, especially on cytologic specimens or small bronchoscopic biopsies. Immunohistochemistry may be of help in this setting: available markers include TTF-1 and SP-A, which are markers of lung origin, whereas there are no good markers of intestinal origin, besides cytokeratin 7 and 20 coexpression pattern, which is not very specific. The nuclear CDX-2 transcription factor, which is the product of a homeobox gene necessary for intestinal organogenesis, is expressed in normal colonic epithelia and most colorectal adenocarcinomas, and could potentially be of diagnostic usefulness. Our aim was to investigate CDX-2 immunohistochemical expression using a new monoclonal antibody and to verify if CDX-2 can be a reliable marker to identify the colorectal origin of lung metastases. CDX-2 expression was evaluated in formalin-fixed, paraffin-embedded samples of normal adult human tissues (50 samples) and in 299 surgically resected carcinomas of different origins, including 125 non-lung adenocarcinomas, 117 primary lung tumors, 5 mesotheliomas, and 52 adenocarcinomas metastatic to the lung. CDX-2 was also evaluated on a series of 20 bioptic and 10 cytologic specimens (5 cases of colorectal metastases to the lung, 5 cases of metastases from other organs, and 10 primary lung adenocarcinomas). In normal tissues CDX-2 immunoreactivity was observed only in ileal and colorectal epithelia. CDX-2 was expressed in almost all primary and metastatic CRC (88 of 90) and was never observed in primary lung tumors. CDX-2 was also expressed in a limited group of adenocarcinomas of other sites (gastric, biliopancreatic, and mucinous ovarian adenocarcinomas). CDX-2 could be easily detected in all bioptic and cytologic samples of CRC metastases. CDX-2 is a reliable, specific, and sensitive immunohistochemical marker of normal and neoplastic intestinal epithelium. CDX-2 can be easily applied to routine histologic and cytologic material and is therefore a useful marker in the differential diagnosis of primary versus metastatic adenocarcinomas in the lung, and among metastases from an unknown primary, supports intestinal origin. PMID- 12548160 TI - Expression of calretinin, thrombomodulin, keratin 5, and mesothelin in lung carcinomas of different types: an immunohistochemical analysis of 596 tumors in comparison with epithelioid mesotheliomas of the pleura. AB - Several immunohistochemical markers, among them calretinin, thrombomodulin (CD141), keratin 5, and mesothelin, have been documented or suggested as useful markers for positive identification of mesothelioma and to differentiate it from pulmonary adenocarcinoma; numerous studies have documented their variable specificity. However, expression of these markers in other types of lung carcinomas has not been systematically explored, although these tumors can enter in the differential diagnosis of mesothelioma. In this study we immunohistochemically evaluated 596 lung carcinomas of different types for the four above-mentioned mesothelioma markers, all of which reacted with a great majority of epithelioid mesotheliomas studied for comparison. Calretinin expression was common in giant cell carcinomas (67%), small cell carcinomas (49%), and large cell carcinomas (38%), whereas it was rare in usual adenocarcinomas but slightly more common in those with neuroendocrine differentiation (11% and 17%, respectively). Thrombomodulin was present in all keratinizing squamous carcinomas and the great majority (87%) of nonkeratinizing tumors in a membrane-staining pattern. It was moderately common in small cell (27%) and large cell carcinomas (25%) but relatively rare in adenocarcinomas (13%). Keratin 5 was expressed in all keratinizing and the great majority (87%) of nonkeratinizing squamous carcinomas, and a majority of large cell carcinomas (56%) and some small cell carcinomas (27%). It was rare in acinar adenocarcinomas (12%) and absent in those with neuroendocrine differentiation. Mesothelin was present in more than half (53%) of adenocarcinomas and a minority (13%) of large cell carcinomas but was absent in small cell carcinomas. In squamous carcinomas it was more often seen in nonkeratinizing versus keratinizing tumors (31% vs 16%). These results show that each of these "mesothelioma" markers reacts with different subsets of pulmonary carcinomas with a variable frequency; this should be considered when using these markers in the differential diagnosis of thoracic tumors. PMID- 12548161 TI - Microscopic papillary thyroid carcinoma compared with clinical carcinomas by loss of heterozygosity mutational profile. AB - The clinical significance of microscopic papillary thyroid carcinoma (PTCa) is controversial. Many authors think that microscopic PTCa (<1 cm) have the same pathogenetic origin as clinically sized papillary carcinomas (>1 cm). Despite the fact that all clinical risk prognostication schemes have the size of the tumor as a primary category, small tumors do have malignant potential and can metastasize. There is growing evidence that small PTCa have the molecular translocations between the proto-oncogene RET and various activating partner genes that are characteristic of clinically sized PTCa. This study used a microdissection and genotyping assay to study the patterns of loss of heterozygosity of tumor suppressor genes in microscopic and clinically sized PTCa. Our results indicate that all PTCa harbor mutations with similar frequencies and distribution patterns, regardless of the size of the tumor. These data are further evidence that microscopic and clinically sized PTCa are pathogenetically related. PMID- 12548162 TI - Pathologic features of uteri and leiomyomas following uterine artery embolization for leiomyomas. AB - The objectives of this study were to identify the presence/absence and location of any embolic material and to describe the morphologic appearance of the leiomyoma and adjacent tissues of cases undergoing surgical intervention following uterine artery embolization (UAE) for leiomyomas. A total of 555 women underwent UAE using polyvinyl alcohol particles (PVA) in a multicenter clinical trial. The histopathologic slides from 17 of 18 women who subsequently underwent myomectomy or hysterectomy in the follow-up period (median 8.2 months) were reviewed without knowledge of the indication for surgery or time elapsed since UAE. The presence/absence and distribution of PVA emboli, associated inflammatory response, and necrosis were noted. Necrosis of leiomyoma(s) was classified as hyaline-type, coagulative tumor cell necrosis, and/or acute suppurative necrosis. In all cases PVA emboli were identified within smooth muscle tumors of the uterine body, its periphery, cervix, uterine body, myometrium, and/or the adnexa. A florid foreign body giant cell type of chronic inflammatory reaction was seen within 1 week of UAE and persisted with visible PVA for up to 14 months post-UAE. Typically, post-UAE leiomyomas showed hyaline-type, but rarely coagulative tumor cell necrosis and acute suppurative necrosis could be seen as well. Five of eight cases coming to surgery for complications showed necrotizing endomyometritis with tissue infarction. PVA particles are recognizable in post-UAE specimens. Leiomyoma necrosis is typically of the hyaline type; coagulative tumor cell necrosis was rarely seen. In some cases with complications, uterine and/or cervical necrosis occurred. The applicability of these findings for UAE patients who have been successfully treated and not resected is uncertain. PMID- 12548163 TI - CD10 expression in epithelial tissues and tumors of the gynecologic tract: a useful marker in the diagnosis of mesonephric, trophoblastic, and clear cell tumors. AB - We tested 417 cases of formalin-fixed, paraffin-embedded normal or hyperplastic gynecologic tissues as well as neoplasms involving the gynecologic tract with a monoclonal antibody against CD10 (clone 56C6), with special emphasis on epithelial and epithelial-like structures and tumors. CD10 was always expressed in mesonephric remnants (mesonephric remnants of the uterine cervix, epoophoron, rete ovarii) and tumors (mesonephric adenocarcinoma of the uterine cervix, tumors of wolffian origin of the broad ligament and ovary). CD10 was also positive in the syncytiotrophoblast, cytotrophoblast, and intermediate trophoblast of normal gestations, partial and complete moles, choriocarcinoma, and placental site trophoblastic tumors. Finally, CD10 was positive in several metastatic neoplasms to the gynecologic tract (100% in metastatic renal clear cell and intestinal carcinomas and melanomas). In contrast, CD10 was almost invariably negative in mullerian epithelia of the female genital tract and in their corresponding tumors, with the exception of focal expression found in squamous epithelia and tumors with squamous differentiation. Thus, the expression of CD10 may be useful in the establishing the diagnosis of mesonephric and trophoblastic tumors and in the differential diagnosis between gynecologic clear cell carcinoma (always negative) and metastatic clear cell carcinoma of renal origin. PMID- 12548164 TI - p16INK4a is a useful marker for the diagnosis of adenocarcinoma of the cervix uteri and its precursors: an immunohistochemical study with immunocytochemical correlations. AB - p16 is a tumor suppressor gene that plays a central role in the regulation of the cell cycle. In squamous cervical cancers, overexpression of p16 is induced by HPV and associated with the carcinogenesis of cervical epithelia. The aim of this study was to determine whether immunostaining of p16 is useful in detecting adenocarcinomas of the cervix uteri in histologic and cytologic routine specimens. A total of 45 surgical specimens, including 18 cases of invasive carcinoma, 8 cases of adenocarcinoma in situ, 4 cases of endocervical glandular atypia (cervical glandular intraepithelial neoplasia), and 15 reactive lesions of the endocervical glands were immunostained using a specific anti-human p16 monoclonal antibody (clone E6H4, mtm laboratories AG, Heidelberg, Germany). Furthermore, immunocytochemical analysis was performed on 10 preoperative ThinPrep cytologic samples with abnormal glandular cells and compared with the human papillomavirus status as assessed with the Hybrid Capture II test. p16 was detected immunohistochemically in all 26 cases of adenocarcinoma of the cervix uteri, including 18 invasive and 8 in situ carcinomas. Only a focal expression was evidenced in the four specimens with endocervical glandular atypia, and no reaction was found in reactive lesions. Also, the immunocytochemical analysis on the 10 ThinPrep specimens evidenced a strong expression of p16 in neoplastic endocervical cells. In all cases this was associated with a high-risk HPV positive typing. p16 is a useful marker for the detection of the adenocarcinoma of the cervix uteri and its precursors. The immunocytochemical detection on ThinPrep specimens may contribute to an early detection of endocervical lesions. PMID- 12548165 TI - Effect of time interval on residual disease in breast cancer. AB - The histologic responses of breast tissue to injury are limited. Needle core biopsies of the breast are associated with displacement of tumor cells, and the incidence of tumor displacement decreases as the time interval between needle core biopsy and subsequent excision increases. This suggests that displaced tumor cells are destroyed by reparative processes induced by tissue injury. Residual tumor in a lumpectomy site may also be subjected to the same destructive processes associated with tissue repair. A total of 259 consecutive cases of infiltrating ductal carcinoma with margin-positive lumpectomies and their associated reexcision specimens obtained over a 7-year period were analyzed for the presence, type, and quantity of residual disease. The overall incidence of residual disease was 69%. Residual infiltrating ductal carcinoma was present in 35% of cases, and residual ductal carcinoma in situ was present in 50%. An increased time interval between lumpectomy and reexcision was associated with a decreased incidence of residual infiltrating carcinoma (p <0.0043); this decrease was not found associated with ductal carcinoma in situ. These findings suggest that the host response to injury may destroy residual infiltrating carcinoma cells in some margin-positive cases. PMID- 12548166 TI - Beta defensin-1, parvalbumin, and vimentin: a panel of diagnostic immunohistochemical markers for renal tumors derived from gene expression profiling studies using cDNA microarrays. AB - The common histopathologic subtypes of renal epithelial neoplasms include conventional, or clear cell, renal cell carcinoma (RCC), papillary RCC, chromophobe RCC, and renal oncocytoma. These subtypes differ clinically and pathologically, making accurate classification important. However, this differential diagnosis can be challenging because of overlapping morphology, suggesting a potential utility for ancillary immunohistochemical markers. We used cDNA microarrays to identify candidate markers for distinguishing renal tumor subtypes. In this report we validated differential expression of three candidate markers, beta defensin-1, parvalbumin, and vimentin, and evaluated the use of this immunohistochemical panel as a potential diagnostic tool. Consistent with our cDNA microarray data, chromophobe RCCs and oncocytomas exhibited similar expression profiles: 8 of 8 examples of each subtype were immunohistochemically positive for beta defensin-1 and parvalbumin and negative for vimentin (sensitivity 100%, specificity 100%); 4 of 7 papillary RCCs were positive for beta defensin-1, parvalbumin, and vimentin (sensitivity 57%, specificity 97%); and 22 of 23 conventional RCCs were negative for beta defensin-1, parvalbumin, or both markers (sensitivity 96%, specificity 96%) as well as positive for vimentin (sensitivity 83%). The immunohistochemical panel distinguished renal tumor subtypes with greater specificity than any marker used alone. This work demonstrates that a useful panel of immunohistochemical markers can be derived from differential gene expression profiles determined using cDNA microarrays. PMID- 12548167 TI - Atypia in nonneoplastic prostate glands after radiotherapy for prostate cancer: duration of atypia and relation to type of radiotherapy. AB - It is unknown how long postradiation atypia of benign prostate glands persists and whether the type of radiation is a factor. Forty-four cases consisting of 37 needle biopsies and 7 transurethral resections of the prostate seen in consultation (January 1997 to September 2000) were studied. In two men (5%), the cases were initially sent without a history of radiotherapy. Thirteen patients had minimal cancer (one core) with the remaining showing no residual tumor. Twenty patients were treated with interstitial radiotherapy (brachytherapy) (IRT), 17 with external beam radiation (XRT), and 7 with a combination of both (CT). The time interval between the treatment and tissue sampling ranged from 8 to 72 months (mean 3 months). Slides were reviewed blindly to the type of radiation and the time interval. Radiation-induced atypia in nonneoplastic glands, stromal fibrosis, and vascular changes was scored separately 0-3, with 0 showing no radiation injury and grade 3 showing prominent nuclear atypia, stromal fibrosis, and vascular hyalinization. We derived a combined score for the epithelial atypia from 0 to 300 (% of glands x grade) for each biopsy. For each case, an overall grade from 0 to 3 was given separately for the stromal and vascular changes. Cases were divided into three groups based on time between treatment and biopsy: <24 months (n = 14), between 24 and 48 months (n = 19), and >48 months (n = 11). Because the scores for epithelial atypia with IRT and CT were the same, we combined them into one group. There was more atypia in cases treated with IRT/CT (mean score 190) than XRT (mean score 105) (p <0.00001). There was also a greater degree of stromal fibrosis with IRT/CT than XRT (p <0.04). There was no correlation between the type of treatment and the effect on vessels. There was no change over time in epithelial atypia in men treated with IRT/CT. With XRT, there was less epithelial atypia in cases biopsied >48 months after treatment (mean score 57) compared with those with a shorter interval between biopsy and treatment (mean score 132) (p = 0.02). Radiation atypia in benign prostate glands may persist for a long time after the initial treatment, resulting in a significant pitfall in evaluating prostate biopsies. Prominent radiation effect (100% of the glands showing grade 2 and 3 atypia) was detected up to 72 months in one of the patients treated with IRT. In some cases, the clinician may not be aware of a prior remote history of radiation or does not relay this history to the pathologist. The pathologist must recognize radiation atypia without relying on the clinician to provide this history. The type of radiation therapy (IRT/CT vs XRT) is a major factor in the degree and duration of postradiation epithelial atypia. PMID- 12548168 TI - Noninfectious lung pathology in patients with Crohn's disease. AB - Lung involvement in Crohn's disease is not well characterized. We reviewed our experience with 11 lung biopsies (seven wedge and four transbronchial) from patients with Crohn's disease to study this association further. Negative cultures, special stains for organisms Gomori-methenamine-silver [GMS], acid fast), and polymerase chain reaction for (four cases) were required for inclusion. The group included five women and six men with a mean age of 47 years (range 13-84 years). A diagnosis of Crohn's disease preceded the lung disease in nine patients. In two patients the diagnosis of Crohn's disease followed the diagnosis of their pulmonary disease 1 and 15 months later. Radiologically, eight patients had diffuse infiltrates, two had bilateral nodular infiltrates, and one had a mass. Chronic bronchiolitis with nonnecrotizing granulomatous inflammation was present in four patients, one of whom was taking mesalamine. Two patients had an acute bronchiolitis associated with a neutrophil-rich bronchopneumonia with suppuration and vague granulomatous features. One patient on mesalamine had cellular interstitial pneumonia with rare giant cells. Four patients demonstrated organizing pneumonia with focal granulomatous features, two of whom were taking mesalamine, and one of these two responded to infliximab (anti-tumor necrosis factor) monoclonal antibody therapy. Noninfectious pulmonary disease in patients with Crohn's disease has variable histologic appearances, including granulomatous inflammation and airway-centered disease resembling that seen in patients with ulcerative colitis. Drugs may contribute to pulmonary disease in some patients. PMID- 12548169 TI - Pathogenic Yersinia DNA is detected in bowel and mesenteric lymph nodes from patients with Crohn's disease. AB - Previously, we detected pathogenic (invasive) DNA in the appendices of two patients who later developed Crohn's disease (CD). This subsequent investigation is the first to evaluate a series of specimens from CD patients for the presence of pathogenic DNA. A total of 54 intestinal resection specimens from 52 patients with confirmed CD were evaluated. Lesional tissue was tested by polymerase chain reaction analysis for the presence of genes occurring only in pathogenic Primer pairs are specific for each species, with no known cross reactions with other bacteria. Forty normal bowel specimens, 30 cases of acute appendicitis, and 50 cases of various active colitides served as disease controls. Medical records were reviewed following polymerase chain reaction and histologic evaluation. A total of 17 of 54 resections (31%) contained DNA by polymerase chain reaction. Mesenteric lymph nodes were also positive in eight of these cases. All controls were negative. -positive patients had carried the diagnosis of CD for a median of 10 years before resection (range 1 month to 40 years). We report the first documentation of DNA in a series of CD cases. Further studies are needed, including serial study, over time, of -positive CD patients, as well as prospective studies of newly diagnosed CD patients for evidence of infection. Like previous studies associating infectious organisms with CD, much work remains to elucidate whether the presence of DNA is an epiphenomenon or actually a factor in the pathogenesis of CD. PMID- 12548170 TI - Loss of CD117 (c-kit)- and CD34-positive ICC and associated CD34-positive fibroblasts defines a subpopulation of chronic intestinal pseudo-obstruction. AB - Chronic idiopathic intestinal pseudo-obstruction is a syndrome in which symptoms of intestinal obstruction are present in the absence of mechanical obstruction. Lack of normal pacemaker activity, usually generated by the interstitial cells of Cajal (ICC), could account for the apparent obstruction. ICC are normally located around and between the myenteric plexus ganglia and within muscle and also in the deep muscular plexus of the small bowel and the submuscular plexus of the large intestine, just within the circular muscle. ICC can be demonstrated immunohistochemically with CD117 (c-kit) as well as with CD34, although this is less specific. CD34 also stains a population of fibroblasts that are intimately associated with ICC. To determine whether there is a relative deficiency of ICC and CD34-positive fibroblasts in patients with chronic idiopathic intestinal pseudo-obstruction, tissue from 30 patients of large intestine and eight patients with small intestine pseudo-obstruction was obtained. Controls (large intestinal specimens from 12 patients, small intestinal specimens from six patients) were chosen from resections for Crohn's disease and colorectal neoplasia, both with and without dilatation. Examination of pseudo-obstruction cases identified 10 patients (nine large intestinal and one small intestinal) in which both CD117 and CD34 were absent or severely reduced in all three of the examined areas. In contrast, the control cases, including those with preobstructive dilatation, showed relatively constant ICC staining. These results suggest that there is a proportion of pseudo-obstruction cases in which the ICC are markedly reduced. These results also demonstrate that, in these cases, loss of the kit immunoreactivity is correlated with the loss of CD34 staining: this indicates that both the ICC and the CD34-positive fibroblasts associated with the ICC are absent. These findings will allow surgical pathologists to identify this subpopulation of patients with CIIP using tissue obtained by laparoscopic biopsy of the muscularis propria or surgical resection. PMID- 12548171 TI - Oxyntic mucosa pseudopolyps: a presentation of atrophic autoimmune gastritis. AB - Gastric polyps are often present in the setting of atrophic gastritis. Although the majority of these polyps are nonneoplastic, such as hyperplastic polyps, neoplastic polyps may be present. We discuss nine cases that illustrate an additional nonneoplastic cause of polyps in atrophic gastritis. Specifically, preserved islands of relatively normal oxyntic mucosa in a background of gastric atrophy may appear polypoid endoscopically. The patients (seven women, two men, mean age 64 years) presented with nonspecific abdominal or reflux symptoms (n = 8) and diarrhea (n = 1). Five of five patients tested were confirmed to have hypergastrinemia, and three of three patients tested had antiparietal cell antibodies. Biopsies from the gastric body or fundus of our nine patients showed fragments of atrophic mucosa and separate fragments of preserved oxyntic mucosa. Based upon the histologic characteristics of the atrophic fundic and relatively normal antral biopsies, the gastric atrophy appeared to be of autoimmune-type. The relatively preserved oxyntic glands showed parietal cell hypertrophy and focal mild chronic inflammation. The number of polyps observed endoscopically ranged from less than five to multiple/diffuse. Three patients had persistent nodularities in their stomachs for 1, 3, and 7 years of their follow-up. Our study shows that some patients with atrophic gastritis, autoimmune-type, may present with gastric polyps/nodules that represent relatively preserved oxyntic mucosa. This presentation may be more common than is presently recognized because biopsies of the polyps alone will not show histologic features of atrophic gastritis or reveal the etiology of the polyp itself. Although a limited number of previous studies have suggested this type of polypoid presentation may represent "early" atrophic gastritis, its persistence in three of our patients argues against this hypothesis. PMID- 12548172 TI - Squamous predominance in mixed-epithelial papillary cystadenomas of borderline malignancy of mullerian type arising in endometriotic cysts: a study of four cases. AB - Mixed-epithelial papillary cystadenoma of borderline malignancy of mullerian type (MEBMM) is composed of a mixture of mullerian epithelial types, such as mucinous, serous, endometrioid, and squamous. Four cases of MEBMM with squamous overgrowth (MEBMMSO) were reviewed. The patients' median age was 56 years, and all cases were unilateral. The clinical stages were Ia (two cases), Ic (one case), and IV based on the presence of tumor cells in pleural fluid (one case). No recurrence was seen in three of the cases. In one of those three cases, there was no recurrence after undergoing surgery only; in the other two of those three cases, there was no recurrence after undergoing surgery and receiving postoperative chemotherapy. In the single case that was at stage IV at initial presentation, a recurrent MEBMMSO nodule was found at a second look 17 months after the initial surgery. In terms of gross findings, all of the tumors were cystic with intracystic papillary fronds. In addition, old endometriotic lesions lined the cysts. The tumors were mainly composed of a proliferation of squamous-type epithelium, with minor foci containing a mixture of other mullerian-type epithelia, especially mucinous. Intraepithelial infiltration by neutrophilic leukocytes was prominent. The differential diagnosis of MEBMMSO includes proliferating Brenner tumors. PMID- 12548173 TI - Malignant mesothelioma of the peritoneum presenting as an inflammatory lesion: a report of four cases. AB - Most cases of malignant mesothelioma present with obvious diffuse tumor, and the presence of grossly visible diffuse tumor is usually cited as an important criterion for making the diagnosis. We report four cases of unsuspected malignant mesothelioma of the peritoneum presenting as localized acute inflammatory lesions. The clinical diagnoses were acute appendicitis in two cases, acute cholecystitis in the third case, and incarcerated umbilical hernia in the fourth case. In all cases tumor was not evident at initial surgical exploration or on gross pathologic examination, and the diagnosis was only made on microscopic examination of the resected specimens. All cases showed a tubulopapillary form of epithelial mesothelioma with obvious tissue invasion, but the foci of tumor were too small to be seen grossly or were present deep in fibrous tissue. On follow-up all patients developed grossly evident tumor, and one of these patients is alive without evidence of disease 5 years after presentation. We conclude that peritoneal mesotheliomas may occasionally present as inflammatory processes without grossly evident tumor and can be diagnosed by microscopic findings alone. PMID- 12548174 TI - Sampling error in the standard evaluation of endoscopic colonic biopsies. AB - It is assumed that the diagnostic information in small gastrointestinal mucosal biopsies is obtained regardless of which side of the biopsy is sectioned. Based on this assumption, mucosal biopsies of the colon are embedded randomly with respect to the side of the biopsy that faces the microtome blade. Our hypothesis is that reorienting ("flipping") the biopsy specimen 180 degrees and sectioning the opposite side might provide new diagnostic information in cases with no specific histopathologic diagnosis. Sixty-one consecutive cases with a clinical impression of a polyp and no histologic abnormality on three hematoxylin and eosin-stained slides (6-15 levels) were selected. Formalin-fixed, paraffin embedded tissue was reoriented, and three additional hematoxylin and eosin stained slides (6-15 levels) were evaluated. The side of the specimen initially sectioned was arbitrarily designated "A," and the side of the specimen sectioned after reembedding was designated "B." The slides were sequentially numbered 1-3, and the first slide on which the new diagnosis was evident was recorded. New diagnoses were made for 19 of 61 (31.1%) specimens from a total of 51 patients and included tubular adenomas (12 of 61; 19.7%) and hyperplastic polyps (7 of 61; 11.5%). In 17 of 19 specimens (89.5%), the new diagnosis was evident on the first slide. Reembedding endoscopic colonic biopsies facilitates rendering a definitive diagnosis in cases of discordance between the clinical impression and the histologic impression. Routine examination of at least two sides of randomly oriented biopsy material could provide additional diagnostic, and sometimes critical, information. PMID- 12548175 TI - Histiocytic sarcoma involving the central nervous system: clinical, immunohistochemical, and molecular genetic studies of a case with review of the literature. AB - Histiocytic sarcoma (HS) is a rare disease, and there has been much confusion concerning the diagnostic criteria for this entity. Since immunohistochemical and cytogenetic techniques have become more universally available, many cases initially diagnosed as histiocytic sarcoma have been reclassified as other diseases. We describe a case of HS that presented as a single mass lesion in left occipital lobe. At autopsy the tumor also involved the meninges as a thick exudate. Histologic examination showed numerous large pleomorphic malignant cells with areas of necrosis, numerous neutrophils, and phagocytosis by tumor cells. Immunohistochemically, the tumor cells stained positively with antibodies directed against most histiocytic markers and did not stain with antibodies directed against myeloid markers, dendritic markers, CD30, ALK1, or other lymphoid markers. Molecular cytogenetic analysis showed no rearrangement [i.e. t(2;5) translocation or other variant] by fluorescence in situ hybridization. The T-cell receptor-gamma chain by multiplex polymerase chain reaction showed a polyclonal pattern. No heavy or light chain gene rearrangements were found. To our knowledge, this is the first reported autopsy case of this rare entity primarily involving the brain and meninges. PMID- 12548176 TI - Difference between familial and sporadic medullary thyroid carcinomas. PMID- 12548177 TI - Board certification in anatomic pathology. PMID- 12548178 TI - Risk for subsequent development of breast cancer. PMID- 12548179 TI - Cardiac mucosal changes in a pediatric population. PMID- 12548180 TI - Histopathology of the gastroesophageal junction: a study on 36 operation specimens. PMID- 12548181 TI - Third molar extraction. Current trends. AB - Extraction of third molars is the operation most frequently performed by oral and maxillofacial surgeons. The American National Institute of Health (1979) and the American Association of Oral and Maxillofacial Surgeons (1993) have organised consensus conferences to systematically analyse the scientific evidence and provide official guidelines for the management of third molars. However, some clinical aspects are still controversial: indications for preventive extraction, early extraction, prognosis of impacted teeth, continuous follow-up, relationship with anterior crowding and temporo-mandibular disorders. This article reviews the conclusions from the consensus conferences and integrates them with more recent data. The actual trend seems to justify a more conservative clinical approach rather than an aggressive extraction policy. PMID- 12548182 TI - [An in vitro study of the tensile strength of a new titanium root post with respect to the type of luting cement used]. AB - BACKGROUND: Assessing the retentive capabilities of a new titanium root post (Optipost, Komet-Gebr. Brasseler, GmbH and Co. KG Lemgo, Germany) with a decreasing diameter shape (turret like) and how these can be improved by the choice of fixing cement. METHODS: The tensile strength was assessed in relation to the type of cement used. The following cements were tested: Compolute composite cement (Espe), C/B Crown & Bridge Luting Composite (Bisco), Diamond Link cement (DRM) with Harvard cement as a control. The cementing procedures laid down by the manufacturers were followed in the case of each cement. The results of the traction tests were analysed and compared statistically and a SEM evaluation of the root/cement and cement/post interface was carried out. RESULTS: The results showed that the highest absolute mean value (best performance) was Diamond Link cement (46.48+/-14.31 kg) which also came out best in the statistical analysis (Scheffe Test p=0.007). Analysis of the samples at the SEM confirmed the results of the traction tests. CONCLUSIONS: The characteristic <> form of the Optipost root post and the sanding of the surface would appear to be important factors which, together with the adhesive cementing technique, lead to a high decementing resistance. PMID- 12548183 TI - [Profile Orifice Shapers. A valid instrument also for the root canal preparation of short teeth: proposal for a series modification]. AB - BACKGROUND: The several endodontic instruments, currently used for the canal preparation with the crown-down technique, simplified only in part root canal therapy, but increased the costs. The authors examined the Profile Orefice Shapers Series, planned only for the preparation of the coronal third, and carried out a comparative analysis with other endodontic instruments currently available. METHODS: Eighty-seven teeth with a 19 mm work lenght were selected for this study. Endodontic treatment was performed using exclusively Profile Orefice Shapers under constant irrigation with 2.5% sodium hypoclorite to clean and shape the canal and System B and Obtura II to obturate the root canal system. RESULTS: Postoperative films show an adequate root canal shaping and a complete apical seal. Endodontic therapy was simplified (only 5 instruments to obtain a good canal shaping); clinical time and costs were decreased. CONCLUSIONS: The authors obtained a good shaping and apical seal using the Profile Orefice Shapers in short teeth. The quality of the root canal therapy obtainable with these instruments is confirmed by radiographs. Some modifications are suggested: increase of instruments working length, introduction of 3 extra intermediate instruments and restriction of working part diameter (1 mm) and operative procedures that lead to obtain an excellent and complete use of these instruments. PMID- 12548184 TI - [Decontamination of dental unit waterlines using disinfectants and filters]. AB - Bacterial contamination of the dental unit water system can become a health problem for patients, particularly if they are immunodepressed. The present study has had the purpose of evaluating the effectiveness of methods of chemical decontamination using different disinfectants (peracetic acid, hydrogen peroxide, silver salts, chloramine T, glutaraldehyde T4) and methods of physical decontamination using synthetic membranes for the filtration of water. A preliminary removal procedure of the biofilm present in the waterline has been followed in a dental unit prepared on purpose for the research; subsequently different 2-week long maintenance procedures were applied using disinfectants injected by a pump and finally the bacterial contamination of the water flowing from the waterline was evaluated. The physical decontamination was performed using 0.22 mm membrane filters, which have been installed also in another dental unit, and the filtered water was analyzed to detect bacterial contamination. The preliminary procedure of biofilm removal succeeded obtaining germ-free water. Among the disinfectants used for the maintenance of the water quality only glutaraldehyde T4 was able to reduce the bacterial contamination under the limit suggested by the ADA. The membrane filter system was not able to purify the water, but when a disinfectant (peracetic acid) was used in the last part of the waterline good results were obtained. At present no decontamination system of dental waterline is available, and glutaraldehyde T4 seems to be the best disinfectant only if integrated with periodic biofilm removal for the maintenance of the water quality. PMID- 12548186 TI - A teachable moment: research ethics revisited. PMID- 12548187 TI - Antibody reactivity to HPV E6 and E7 oncoproteins and early diagnosis of invasive cervical cancer. PMID- 12548188 TI - Use of anthelminthic drugs during pregnancy. PMID- 12548189 TI - Hysteroscopic endometrial resection versus laparoscopic supracervical hysterectomy for menorrhagia: a prospective randomized trial. AB - OBJECTIVE: This study was undertaken to compare the relative efficacy and safety of hysteroscopic endometrial resection and laparoscopic supracervical hysterectomy in the treatment of abnormal uterine bleeding. STUDY DESIGN: One hundred eighty-one patients affected by menometrorrhagia and unresponsive to medical treatment agreed to be randomized to either laparoscopic supracervical hysterectomy or hysteroscopic endometrial ablation. They were monitored for 2 years to evaluate perioperative and postoperative outcomes, resolution of symptoms, and patient satisfaction. RESULTS: Duration of hospitalization, period of convalescence, perioperative complications, and resumption of normal activity were similar between the two groups. Operative time was significantly shorter in the hysteroscopic group, but patient satisfaction was significantly higher in the laparoscopic group. CONCLUSION: For the treatment of menorrhagia, hysterectomy has the distinct advantage of being curative but the disadvantage of being more invasive than the hysteroscopic approach. However, laparoscopic supracervical hysterectomy preserves the curative effect of hysterectomy without its increased surgical invasiveness, as suggested by the current study. PMID- 12548190 TI - Gestation-related and betamethasone-induced changes in 11beta-hydroxysteroid dehydrogenase types 1 and 2 in the baboon placenta. AB - OBJECTIVE: We determined developmental and labor-related changes in 11beta hydroxysteroid (HSD) 1 and 2 expression in baboon placentas during the final third of gestation and labor. We examined whether maternal glucocorticoid administration alters placental 11beta-HSD 2 expression. STUDY DESIGN: Maternal and fetal plasma cortisol concentrations were measured in five animals. Types 1 and 2 11beta-HSD messenger RNA (mRNA) and protein in placentas obtained at 121 to 185 days' gestation (dGA, term approximately 185 dGA, n = 16), during labor between 141 and 193 dGA (n = 8), and after maternal administration of four doses of 87.5 microg/kg betamethasone (n = 5) at 12-hour intervals at 121 to 135 dGA were analyzed by Northern and Western blot. RESULTS: Cortisol levels were higher in maternal plasma than fetal (4-fold, P <.mob031). Placental 11beta-HSD 2 mRNA and protein decreased after 0.9 gestation (P <.001). 11beta-HSD 1 mRNA remained unchanged. There was no effect of labor on placental 11beta-HSD 1 and 2 mRNA and protein levels. Maternal betamethasone administration dramatically increased (P <.05) 11beta-HSD 2 mRNA as well as protein without effect on 11beta-HSD 1 mRNA and protein expression. CONCLUSIONS: The late-gestation baboon maternal plasma cortisol concentration is four times the fetal plasma concentration. Decreased placental 11beta-HSD 2 may enhance maternal cortisol passage to the fetus at the end of gestation, thereby contributing to cortisol-mediated changes within the placenta and cortisol in fetal plasma at this stage of fetal development. The positive effect of betamethasone on placental 11beta-HSD 2 induction further suggests an ability of the placenta to regulate glucocorticoid transfer in the presence of elevated maternal glucocorticoid. PMID- 12548192 TI - Radical trachelectomy and pelvic lymphadenectomy with uterine preservation in the treatment of cervical cancer. AB - OBJECTIVE: The purpose of this study was to determine whether radical trachelectomy, combined with pelvic lymphadenectomy, can be a feasible method for the treatment of early-stage cervical carcinoma in women who want to preserve their fertility. STUDY DESIGN: From January 1, 1995, through December 31, 1999, 12 women with stage I carcinoma of the cervix were scheduled to undergo radical trachelectomy and pelvic lymphadenectomy of the intact uterus as treatment. The procedure was abandoned in 2 women because of endometrial extension of the cancer. Surgical margins were clear in all other women. No lymph node metastases were encountered. The proximal cervical remnant was reinforced in 10 women. RESULTS: Hospitalization ranged from 2 to 8 days (mean, 3.2 days). Estimated blood loss averaged 203 mL (range, 50-600 mL). Complications included 2 intraoperative cystotomies and 1 pelvic hematoma. Four pregnancies have occurred, with 2 third-trimester deliveries and 2 preterm losses at 24 and 26 weeks of gestation, respectively. The follow-up period has ranged from 28 to 84 months (mean, 47.6 months). CONCLUSION: Radical trachelectomy, combined with pelvic lymphadenectomy, can be a feasible method of treatment for early-stage cervical carcinoma in women who want to preserve their fertility. PMID- 12548191 TI - Differential effects of maternal betamethasone and cortisol on lung maturation and growth in fetal sheep. AB - OBJECTIVES: We asked whether maternal or fetal cortisol treatments induced fetal lung maturation and growth restriction compared with betamethasone and whether medroxyprogesterone (MPA) influenced lung maturation. STUDY DESIGN: Pregnant sheep were randomized to receive MPA or saline solution at 100 days of gestation. At 117 days of pregnancy, ewes were randomized to receive maternal or fetal treatments with 0.5 mg/kg betamethasone, four injections of hydrocortisone given over 4.5 hours or saline solution. Body weight and lung maturation were evaluated at 125 days. RESULTS: Maternal betamethasone decreased in birth weight by 16% and induced lung maturation. Fetal cortisol or betamethasone induced only lung maturation. Maternal cortisol caused neither growth restriction nor lung maturation. MPA did not alter lung function in control lambs or in betamethasone treated preterm lambs. CONCLUSIONS: Maternal cortisol treatments had no apparent effects on the fetus, suggesting that growth restriction caused by betamethasone may result from unidentified effects on the maternal-placental unit. PMID- 12548193 TI - Comparison of the growth and hydrogen peroxide production by vaginal probiotic lactobacilli under different culture conditions. AB - OBJECTIVE: The purpose of this study was to determine the optimal conditions for the growth and hydrogen peroxide production by vaginal lactobacilli. STUDY DESIGN: Two vaginal lactobacillus strains were cultured under different growth conditions (temperature, pH, agitation, and growth media) with a fractional factorial experimental design. RESULTS: The optimal growth conditions of Lactobacillus paracasei CRL (Centro de Referencia para Lactobacilus Culture Collection) 1289 and Lactobacillus crispatus CRL 1266 were temperature 37 degrees C and initial pH 6.5, nonagitated cultures, in autolysat de levure; peptone, tryptone, Tween 80 et glucose (LAPTg) broth for L paracasei or in de Man-Rogosa Sharpe (MRS) and LAPTg broths for L crispatus. The hydrogen peroxide production, detected only in agitated cultures, was higher at 37 degrees C and pH 6.5. The oxidative metabolite produced the self-inhibition of the lactobacilli growth. In mixed cultures of lactobacilli and Staphylococcus aureus under the optimal conditions of hydrogen peroxide production, the pathogen growth was inhibited. CONCLUSION: The results provided information about the factors that affect the hydrogen peroxide production and about the most favorable conditions with which to obtain the highest biomass in the shortest possible time. PMID- 12548194 TI - Increased risk of thyroid cancer among women with hysterectomies. AB - OBJECTIVE: Hysterectomy with bilateral oophorectomy has been suggested to increase the risk of thyroid cancer. We studied the relationship between hysterectomy and thyroid cancer in a population-based setting in Finland. STUDY DESIGN: Women undergoing hysterectomy between 1986 and 1995 (n = 17,900) were identified from the National Hospital Discharge Registry. The cohort was followed up through the Finnish Cancer Registry until 1997. RESULTS: There were 118 cases of thyroid cancer diagnosed, 103 papillary and 15 follicular or medullar type. The incidence for thyroid cancer was significantly elevated (standardized incidence ratio [SIR] 1.38, 95% CI 1.15-1.64). The increase in the incidence of thyroid cancer was not dependent on the extent of operation but on the length of follow-up. Thyroid cancer incidence was increased 0.5 to 1.4 years after hysterectomy (SIR 2.00, 95% CI 1.31-2.93), but decreased thereafter (SIR 1.30, 95% CI 0.99-1.67). Hysterectomy with and without oophorectomy was associated with a similar increase in the incidence of thyroid cancer. CONCLUSION: Women who have undergone hysterectomy have an increased risk of thyroid cancer during the first 2 years after the operation. Thyroid cancer and bleeding disorders may share a common background. PMID- 12548196 TI - Genotyping of 22 human papillomavirus types by DNA chip in Korean women: comparison with cytologic diagnosis. AB - OBJECTIVE: More sensitive and reliable methods than individual testing (such as polymerase chain reaction, restriction fragment length polymorphism, and Southern blot) should be developed as screening tools for the detection of latent human papillomavirus. Today, the new Bethesda system recommends human papillomavirus testing as an adjuvant to the conventional Papanicolaou smear for more comprehensive identification of women at certain risk of cervical neoplasia. We performed human papillomavirus genotyping with the newly designed human papillomavirus DNA chip, which is based on polymerase chain reaction for high throughput screening power, and compared the results with the results of a Papanicolaou smear according to the new Bethesda system. STUDY DESIGN: Polymerase chain reaction amplifications of the human papillomavirus L1 region from biologic samples were hybridized to silanized glass slides by a microarrayer, which comprised 22 specific oligonucleotide probes to their genotypes, consisting of 15 high-risk and 7 low-risk types. Two cervical cancer cell lines and 20 plasmids that contained each type of the human papillomavirus whole genome were used for the evaluation of this method; in all cases, the cancer cell lines and plasmids showed clear positive signals on their corresponding positions. A comparative study that used 685 cervicovaginal swabs was performed by human papillomavirus DNA chip microarray together with Papanicolaou diagnosis. RESULTS: Human papillomavirus was identified as positive in 31.9% of the 414 control samples and in 78.6% of the 271 neoplastic lesions. The major prevailing human papillomavirus genotypes were human papillomavirus types 16, 58, and 18, in descending order of incidence (average overall, 78.8%). Almost all of the remaining cases were comprised of human papillomavirus types 39, 52, 56, and 51. The frequency of multiple infection of human papillomavirus was highest in low-grade squamous intraepithelial lesion but was lowest in squamous cell carcinoma. All cases that exhibited infection of single human papillomavirus type 58 were squamous cell carcinoma. CONCLUSION: Human papillomavirus types 16, 18, and 58 were confirmed to be major causative factors for cervical carcinogenesis. Low-grade squamous intraepithelial lesion is a heterogeneous entity that is composed of different human papillomavirus subtypes and prevails in younger women (<40 years old). The human papillomavirus chip has potential use as a high-throughput screening test. PMID- 12548195 TI - Evaluation of antibody response to human papillomavirus early proteins in women in whom cervical cancer developed 1 to 20 years later. AB - OBJECTIVE: Infection with oncogenic human papillomaviruses (HPVs) is the most important cause of cervical cancer worldwide. After infection there is a long latency period of at least 10 to 15 years during which cervical cancer develops in a small proportion of originally infected women. Up to 50% of these women have at diagnosis antibodies to the HPV oncoproteins E6 and E7, which are rarely found among healthy women. Our purpose was to evaluate whether antibodies to HPV16 and HPV18 E6 and E7 proteins are useful for early diagnosis of cervical cancer by measuring the antibody response in women in whom cervical cancer later developed. STUDY DESIGN: A joint serum bank of 550,000 Swedish, Norwegian, and Finnish women was followed up for 0.5 to 20 years, after which 178 invasive cervical carcinoma (ICC) cases, 150 of whom had squamous cell carcinoma (SCC), and 527 controls were identified. Antibodies to HPV16 and HPV18 E6 and E7 proteins were determined by tag enzyme-linked immunoassays. RESULTS: HPV16/18 E6 and E7 antibodies were detected infrequently (7.0%) in women in whom SCC later developed and yielded a moderately increased estimate of associated relative risk (odds ratio 2.7, 95% CI 1.1-6.4). Sensitivity of the combined antibody tests for the detection of occult SCC varied between 6% and 14% but was not related to time lag between serum sampling and cancer diagnosis. CONCLUSION: HPV16/18 E6 and E7 antibody responses are not sensitive markers of occult cervical cancer. PMID- 12548197 TI - Pregnant adolescents at risk: sexual behaviors and sexually transmitted disease prevalence. AB - OBJECTIVE: The purpose of this study was to determine the level of high-risk sexual behaviors and the prevalence of chlamydia and gonorrhea among pregnant adolescents. STUDY DESIGN: These analyses used data from 203 pregnant and 209 nonpregnant adolescents who were recruited from public health clinics. Data sources included interviewer-administered questionnaires, ligase chain reaction tests for chlamydia/gonorrhea in the third trimester of pregnancy, and state health department reports of chlamydia/gonorrhea. Statistical analyses included logistic regression. RESULTS: Pregnant adolescents were significantly more likely to have not used condoms during sexual intercourse in the past 30 days compared with nonpregnant adolescents, although other sexual risk behaviors were reduced. Nineteen percent of pregnant adolescents had chlamydia or gonorrhea diagnosed during the pregnancy. CONCLUSION: Pregnant adolescents have high levels of sexually transmitted diseases during pregnancy, and many adolescents use condoms inconsistently. Prenatal care providers may be in a unique position to decrease sexually transmitted diseases among pregnant adolescents by encouraging the reduction of risk behaviors and aggressively screening for sexually transmitted diseases as part of routine prenatal care. PMID- 12548198 TI - Subcutaneous drainage versus no drainage after minilaparotomy in gynecologic benign conditions: a randomized study. AB - OBJECTIVE: The purpose of this study was to determine the outcomes of subcutaneous drainage in patients who are affected by benign gynecologic diseases that are treated by a Kustner's minilaparotomy incision. STUDY DESIGN: We performed a randomized controlled study with 72 patients who underwent operation for benign gynecologic disease. Patients were assigned intraoperatively to two groups: group A (36 patients) had a closed drainage system and group B (36 patients) had no drainage. Historic data, clinical and surgical characteristics, and perioperative complications were recorded prospectively. Postoperative care data, surgical incision complications, and hospital stay were analyzed. Patients were to be seen 4 weeks after the operation to assess wound healing. RESULTS: No patients had intraoperative complications or blood transfusions. A significant difference was observed between the drainage versus no-drainage groups regarding postoperative complications and hospital stay (P =.001, both groups). No patients had surgical complications at 4 weeks after the last operation. CONCLUSION: Our data suggest that routine prophylactic subcutaneous drainage is indicated in patients who undergo Kustner's minilaparotomy incision for benign gynecologic operation. Drains significantly prevent wound complications and reduce hospital stay. PMID- 12548199 TI - Expression, menstrual cycle-dependent activation, and bimodal mitogenic effect of transforming growth factor-beta1 in human myometrium and leiomyoma. AB - OBJECTIVE: Transforming growth factor-beta1 is the prototype of a bimodal regulator of cell growth, which can either inhibit or stimulate the proliferation of smooth muscle cells. Part of transforming growth factor-beta1-mediated stimulation of growth is associated with the increased production of platelet derived growth factor. The conversion of latent-to-active transforming growth factor-beta provides a pivotal mechanism for the regulation of the biologic activity of transforming growth factor-beta. We investigated the differential expression and production of the active form of transforming growth factor-beta1 in the myometrium and leiomyoma throughout the menstrual cycle. We also studied the mitogenic effects of transforming growth factor-beta1 and platelet derived growth factor on myometrial and leiomyoma cells in culture. STUDY DESIGN: Myometrium and leiomyoma tissue pairs were obtained from 28 women who underwent hysterectomy. Total RNA from each tissue was extracted, and Northern blot analysis was performed for the detection of TGF-beta1 messenger RNA. Active and total transforming growth factor-beta1 protein was quantified with enzyme-linked immunosorbent assay. Cell proliferation of cultured human myometrial and leiomyoma cells that are treated with TGF-beta1 (0.01-1 ng/mL), anti-transforming growth factor-beta antibody (0.01-10 ng/mL), or platelet-derived growth factor (10 ng/mL) was assessed by the [(3)H]thymidine incorporation method. RESULTS: Overall, the transforming growth factor-beta1 messenger RNA level in myometrial samples was 1.2-fold higher than in the leiomyoma samples (P <.05). Active transforming growth factor-beta1 protein levels in follicular and luteal phase myometrial and leiomyoma samples were significantly greater than the levels in samples from women with atrophic endometrium (P < 0.05). Transforming growth factor-beta1, at low concentrations (0.01 ng/mL), induced an increase in cell proliferation (2- to 3-fold; P <.05). When cells were treated with anti transforming growth factor-beta antibody, there was a larger magnitude of increase observed (7- to 20-fold; P <.05). Platelet-derived growth factor (10 ng/mL) consistently increased the rate of cell proliferation both in myometrium and leiomyoma cells (5- to 6-fold; P <.05). CONCLUSION: Levels of active transforming growth factor-beta1 that were produced in follicular and luteal phases indicate a stimulatory role for ovarian hormones. The finding that transforming growth factor-beta1, only at low concentrations, stimulates cell proliferation mainly in leiomyoma cells is in agreement with the bimodal and dose dependent effects of transforming growth factor-beta1 that is observed in smooth muscle cells of other tissues. The persistent and high rate of cell proliferation with platelet-derived growth factor suggests that the growth stimulatory effect of transforming growth factor-beta1 may be mediated through its up-regulatory effect on platelet-derived growth factor. PMID- 12548200 TI - Chorion-induced myometrial relaxation is mediated by large-conductance Ca2+ activated K+ channel opening in the guinea pig. AB - OBJECTIVE: We previously demonstrated that chorion releases a factor that inhibits both spontaneous and oxytocin-stimulated myometrial contractility. Here, we investigate the mechanism of action of this unidentified substance. STUDY DESIGN: Myometrial strips from pregnant guinea pigs were mounted in an organ bath and contractility stimulated with oxytocin. RESULTS: Guinea pig chorion produced a time-dependent decrease in oxytocin-induced myometrial contractility. The ability of the chorion to reduce contractility was unaltered by inhibiting chorionic synthesis of either nitric oxide (N [omega]-nitro-L-arginine), carbon monoxide (tin-protoporphyrin), prostaglandins (indomethacin), or the myometrial cyclic guanosine monophosphate pathway (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalime-1 one and Rp-8Br-cGMP). In contrast, iberiotoxin, an inhibitor of large conductance Ca(2+)-activated K(+) channels reduced the quiescent effect of chorion by 40%; in contrast, inhibition of adenosine triphosphate-sensitive (glibenclamide) and voltage-gated K(+) channels (4amynopyridine) had no effect. CONCLUSION: Chorion induced relaxation of oxytocin-stimulated myometrial contraction is, in great part, the product of a paracrine substance that opens myometrial large conductance Ca(2+)-activated K(+) channels. PMID- 12548201 TI - Reduced vaginal bleeding in postmenopausal women who receive combined norethindrone acetate and low-dose ethinyl estradiol therapy versus combined conjugated equine estrogens and medroxyprogesterone acetate therapy. AB - OBJECTIVE: The purpose of this study was to compare the effects on vaginal bleeding patterns of continuous combined hormone replacement therapy with norethindrone acetate and ethinyl estradiol versus conjugated equine estrogens and medroxyprogesterone acetate. STUDY DESIGN: Three hundred fifty-seven postmenopausal women were selected randomly (in a blinded manner) to 12 months of treatment with 1 mg norethindrone acetate/5 microg ethinyl estradiol, placebo, or open-label 0.625 mg conjugated equine estrogens/2.5 mg medroxyprogesterone acetate (conjugated equine estrogens/medroxyprogesterone acetate [CEE/MPA]; Prempro). The incidence and duration of vaginal bleeding were assessed throughout the study. Statistical analyses used Cochran-Mantel-Haenszel methodology and analysis of variance. RESULTS: At 3 months, 1 mg norethindrone acetate/5 microg ethinyl estradiol therapy reduced the incidence of bleeding (12% vs 23%; P <.029) and bleeding and/or spotting (22% vs 44%; P <.001), compared with conjugated equine estrogens/medroxyprogesterone acetate therapy. The mean duration of bleeding and bleeding and/or spotting were also reduced with 1 mg norethindrone acetate/5 microg ethinyl estradiol therapy versus conjugated equine estrogens/medroxyprogesterone acetate (P =.004 and P <.001, respectively). The incidence of cumulative amenorrhea at every monthly interval was significantly better with 1 mg norethindrone acetate/5 microg ethinyl estradiol therapy versus conjugated equine estrogens/medroxyprogesterone acetate therapy (P <.05). Associated adverse event (ie, headache, breast pain) incidence rates were similar in the 2 active treatment groups. CONCLUSION: The 1 mg norethindrone acetate/5 microg ethinyl estradiol therapy provides significantly better control of vaginal bleeding than conjugated equine estrogens/medroxyprogesterone acetate therapy at all time points investigated in this 12-month study. PMID- 12548202 TI - High cumulative incidence of uterine leiomyoma in black and white women: ultrasound evidence. AB - OBJECTIVE: Uterine leiomyoma, or fibroid tumors, are the leading indication for hysterectomy in the United States, but the proportion of women in whom fibroid tumors develop is not known. This study screened for fibroid tumors, independently of clinical symptoms, to estimate the age-specific proportion of black and white women in whom fibroid tumors develop. STUDY DESIGN: Randomly selected members of an urban health plan who were 35 to 49 years old participated (n = 1364 women). Medical records and self-report were used to assess fibroid status for those women who were no longer menstruating (most of whom had had hysterectomies). Premenopausal women were screened by ultrasonography. We estimated the age-specific cumulative incidence of fibroid tumors for black and white women. RESULTS: Thirty-five percent of premenopausal women had a previous diagnosis of fibroid tumors. Fifty-one percent of the premenopausal women who had no previous diagnosis had ultrasound evidence of fibroid tumors. The estimated cumulative incidence of tumors by age 50 was >80% for black women and nearly 70% for white women. The difference between the age-specific cumulative incidence curves for black and white women was highly significant (odds ratio, 2.9; 95% CI, 2.5-3.4; P <.001). CONCLUSION: The results of this study suggest that most black and white women in the United States develop uterine fibroid tumors before menopause and that uterine fibroid tumors develop in black women at earlier ages than in white women. PMID- 12548203 TI - Procedures for pelvic organ prolapse in the United States, 1979-1997. AB - OBJECTIVE: The objective of our study was to describe national rates of surgery for pelvic organ prolapse. STUDY DESIGN: We used the National Hospital Discharge Survey, a federal database that samples inpatient hospitals in the United States. Data from 1979 to 1997 were analyzed for diagnoses and procedures coded with the ICD-9-CM classification system. Age-adjusted rates were calculated by using the 1990 census population and compared for change over time by using the Score test for linear trend. RESULTS: Approximately 200,000 women undergo inpatient procedures for prolapse in the United States each year, ranging from 165,000 in 1988 to 226,000 in 1979. There was a significant decrease in the overall age adjusted rate of procedures performed over the study period, from 2.2 to 1.5 procedures per 1000 women (P =.01). The rate of prolapse procedures for women younger than 50 years decreased by more than half (1.9 to 0.8 per 1000 women, P <.001), whereas the rate remained stable in women who were aged 50 years and older (2.7 to 3.3 per 1000 women, P =.5). CONCLUSION: Prolapse procedures are performed very frequently in the United States, although the rate has decreased slightly over time. PMID- 12548204 TI - Anatomic variations in the levator ani muscle, endopelvic fascia, and urethra in nulliparas evaluated by magnetic resonance imaging. AB - OBJECTIVE: The purpose of this study was to develop a system to quantify interindividual variation in the appearance of continence system structures in normal continent nulliparous women. STUDY DESIGN: Magnetic resonance imaging (1.5 T) was performed in 20 healthy continent nulliparous women (mean age, 30.1 +/- 5.1 years) with normal pelvic organ support and urodynamics. Morphometric measurements of the levator ani muscle, endopelvic fascia, and urethra were performed. RESULTS: The ratio of the maximum-to-minimum measured values shows that 2- to 3-fold differences occur in distance, area, or volume measures of continence system morphologic features. The mean urogenital hiatus area was 15.2 +/- 2.9 cm(2) in women without a visible connection of the levator ani muscle to the pubic bone (4/20 women) and 12.3 +/- 2.4 cm(2) in women with an levator ani muscle-pubic bone connection (16/20 women, P =.05). CONCLUSION: Considerable variation that was not attributable to limitations of the measuring technique that was used occurs in the size and configuration of the urethral support structures in nulliparous asymptomatic women. PMID- 12548205 TI - Genital tract and plasma human immunodeficiency virus viral load throughout the menstrual cycle in women who are infected with ovulatory human immunodeficiency virus. AB - OBJECTIVE: The purpose of this study was to determine the vaginal, cervical, and plasma viral load through the menstrual cycle in women who are positive for human immunodeficiency virus. STUDY DESIGN: A prospective cohort study was performed on 14 women with ovulatory menstrual cycles who have human immunodeficiency virus. Duplicate cervical and vaginal viral load samples (n = 301) were taken at four stages (menstrual, follicular, periovulatory, and luteal) of two consecutive cycles. RESULTS: Participant characteristics were mean age of 32.7 years, median human immunodeficiency virus helper cell count value of 355, and median plasma viral load of 24,000 copies/mL. Through the menstrual cycle, there was no statistically significant difference in plasma viral load, but there was a significant decrease in genital tract viral load at the periovulatory phase (vagina, P =.018; cervix, P =.007). Vaginal and cervical viral load were correlated (r = 0.582, P <.001). CONCLUSION: Although the plasma viral load remained constant throughout the menstrual cycle, the genital viral load decreased at the periovulatory phase. These results suggest that local factors may affect the genital viral load compartment independent of plasma viral load. PMID- 12548206 TI - Herpes simplex virus type II is not a cofactor to human papillomavirus in cancer of the uterine cervix. AB - OBJECTIVE: Cells that were cotransfected with herpes simplex virus-16 and the herpes simplex virus type 2 Xho -2 DNA induce tumors in nude mice. In a cross sectional study, we investigated the role of herpes simplex virus type 2 as a cofactor to human papillomavirus in cervical cancer. STUDY DESIGN: Cervical cells that were obtained with an endocervical Cytobrush brush (Medscand) from 439 women (50 women with cancer lesions, 65 women with high-grade squamous intraepithelial lesions, 80 women with low-grade squamous intraepithelial lesions, 244 healthy subjects) and DNA that was extracted from 150 cervical cancer biopsy specimens were analyzed with polymerase chain reaction for herpes simplex virus type 2 Xho 2 and Bgl IIC transforming DNA sequences. RESULTS: All 439 cervical samples and 150 cervical cancer biopsy specimens tested negative for herpes simplex virus type 2 Xho -2 and Bgl IIC DNA by polymerase chain reaction. Overall, none of 200 samples (0%) from women with invasive cervical cancer contained herpes simplex virus type 2 Xho -2 or Bgl IIC DNA (95% CI, 0.0-1.8). CONCLUSION: Although herpes simplex virus type 2 Bgl IIN transforms epithelial cells in vitro, it was not detected in cervical cancer specimens. PMID- 12548207 TI - Levels of placenta growth factor in gestational trophoblastic diseases. AB - OBJECTIVE: The aim of the current study was to investigate levels of placenta growth factor in the tissues and sera of the patients with gestational trophoblastic disease and to determine its usefulness for the treatment of gestational trophoblastic disease. STUDY DESIGN: Placenta growth factor concentrations were measured in the tissue homogenates of 12 normal placentas, 33 complete hydatidiform moles, and 6 gestational choriocarcinomas. Serum placenta growth factor levels were determined in 59 women with normal pregnant course, in 30 women with complete hydatidiform mole, in 36 women with persistent gestational trophoblastic disease, and 100 nonpregnant healthy volunteers. RESULTS: Serum and tissue placenta growth factor levels in the patients with mole tended to be decreased compared with the levels in normal pregnancy; the levels were increased significantly in patients with choriocarcinoma. When serum placenta growth factor levels were >20 pg/mL (normal upper limit in nonpregnant women), placenta growth factor-to-human chorionic gonadotropin ratios were increased significantly in patients with persistent gestational trophoblastic disease. CONCLUSION: Serum placenta growth factor levels are not of any predictive value in patients with hydatidiform mole. However, elevated serum placenta growth factor levels with increased placenta growth factor-to-human chorionic gonadotropin ratios are suggestive of persistent gestational trophoblastic disease. PMID- 12548208 TI - Endometritis does not predict reproductive morbidity after pelvic inflammatory disease. AB - OBJECTIVE: We investigated the association between endometritis and reproductive morbidity. STUDY DESIGN: Participants were 614 women in the PID Evaluation and Clinical Health (PEACH) Study with pelvic pain, pelvic organ tenderness, and leukorrhea, mucopurulent cervicitis, or untreated cervicitis. We compared women with endometritis (>or=5 neutrophils or >or=2 plasma cells), Neisseria gonorrhoeae or Chlamydia trachomatis upper genital tract infection (UGTI) or both to women without endometritis/UGTI for outcomes of pregnancy, infertility, recurrent pelvic inflammatory disease (PID), and chronic pelvic pain (CPP), adjusting for age, race, education, PID history, and baseline infertility. RESULTS: Endometritis/UGTI was not associated with reduced pregnancy (odds ratio [OR] 0.8, 95% CI 0.6-1.2) or elevated infertility (OR 1.0, 95% CI 0.6-1.6), recurrent PID (OR 0.6, 95% CI 0.4-0.9), or CPP (OR 0.6, 95% CI 0.4-0.9). PEACH participants with and without endometritis/UGTI had higher age- and race-specific pregnancy rates than 1997 national rates. CONCLUSION: Among women with clinically suspected mild-to-moderate PID treated with standard antibiotics, endometritis/UGTI was not associated with reproductive morbidity. PMID- 12548209 TI - Gentamicin and clindamycin therapy in postpartum endometritis: the efficacy of daily dosing versus dosing every 8 hours. AB - OBJECTIVE: The objective of the study was to evaluate the efficacy of gentamicin and clindamycin given once daily versus the more common 8-hour dosing regimen for the treatment of postpartum endometritis. STUDY DESIGN: In a prospective, placebo controlled, double-blinded study, patients who had postpartum endometritis diagnosed were randomly selected to receive 1.5 mg/kg gentamicin and 900 mg clindamycin phosphate administered every 8 hours versus gentamicin 5 mg/kg and clindamycin phosphate 2700 mg administered as a single-daily dose. The single dose group received an infusion of gentamicin and clindamycin, followed by an administration of intravenous placebo 8 and 16 hours later to maintain blinding. Treatment success was defined as absence of fever 72 hours after initiation of antibiotic therapy. RESULTS: One hundred ten patients were enrolled. The daily dose group (n = 55) and the thrice-daily dose group (n = 55) were similar with respect to age, gravidity, parity, gestational age, and maternal weight. Clinical characteristics (including maximum temperature, presence of predelivery chorioamnionitis, white blood cell count, and mode of delivery) were also similar. There was no difference in the mean time from initiation of therapy until becoming afebrile in the daily-dose group (27.4 +/- 24.9 hours) compared with the thrice-daily dose group (32.9 +/- 26.3 hours). Forty-five of 56 (82%) patients in the daily-dose group and 38 of 55 (69%) patients in the thrice-daily dose group had treatment success (P =.12). CONCLUSION: Once-daily dosing with gentamicin and clindamycin in women with postpartum endometritis has a similar success rate as the standard every 8-hour dosing schedule. PMID- 12548210 TI - Defecation in utero: a physiologic fetal function. AB - OBJECTIVE: The objective of this study was to investigate the occurrence of in utero defecation as a normal function in the human fetus. STUDY DESIGN: The anuses of 240 fetuses were studied sonographically between weeks 15 and 41 of gestation. Fetal defecation was defined as the expulsion of rectal contents through the anus into the amniotic fluid. The diameter and area of the anus were measured sonographically at times of maximum anal aperture. RESULTS: One or more defecations were documented in all fetuses. The frequency of defecations was highest between week 28 and 34 of gestation. CONCLUSION: This study confirms that defecation in utero is a normal function and supports the view that the evacuation of rectal contents into the amniotic fluid is no departure from normal fetal physiologic behavior. PMID- 12548211 TI - Effects of colony-stimulating factors on the secretion of platelet-activating factor acetylhydrolase by human decidual macrophages. AB - OBJECTIVE: The aim is to clarify the role of platelet-activating factor and colony-stimulating factors in term and preterm parturition. STUDY DESIGN: Decidual macrophage populations were obtained by an enzymic digestion, Ficoll Paque centrifugation, or flow cytometric sorting. The effects of colony stimulating factors on the platelet-activating factor acetylhydrolase secretion by these cells were examined. RESULTS: Granulocyte-macrophage colony-stimulating factor inhibited the platelet-activating factor acetylhydrolase secretion by decidual macrophages. Granulocyte colony-stimulating factor also decreased the enzyme secretion but at higher concentrations than those required for granulocyte colony-stimulating factor. In contrast, macrophage colony-stimulating factor increased the enzyme secretion. These colony-stimulating factor-induced effects were specifically neutralized by the corresponding antibodies. CONCLUSION: Colony stimulating factors may modulate the local concentration of platelet-activating factor in the decidua via their inhibitory or stimulatory effect on the secretion of platelet-activating factor acetylhydrolase, contributing to the regulation of term and preterm parturition at the maternal-fetal interface. PMID- 12548212 TI - Oral misoprostol or vaginal dinoprostone for labor induction: a randomized controlled trial. AB - OBJECTIVE: The objective of the study was to compare the effectiveness, safety, and side effects of low-dose oral misoprostol with vaginal dinoprostone for cervical ripening and labor induction. STUDY DESIGN: Women with Bishop score 6 or less admitted for labor induction at term were eligible for this randomized controlled trial. Exclusion criteria were multiple pregnancy, breech, fetal distress, or previous uterine scar. The allocation to the oral misoprostol group (20 microg given every 2 hours increased to 40 microg depending on uterine contractions) or to the vaginal dinoprostone group (2 mg twice, 6 hours apart) was contained in a sealed, opaque, and consecutively numbered envelope. RESULTS: Two hundred women (100 in each group) were included. The proportion of vaginal delivery within 24 hours was 56% in the misoprostol group and 62% in the dinoprostone group (relative risk 0.90, 95% CI 0.72-1.14). The risk of cesarean section was 18% and 19%, respectively. The median interval to delivery, calculated from survival analysis, was longer in the misoprostol group (1305 minutes) compared with the dinoprostone group (1080 minutes). The log-rank test was not significant (P =.35). Uterine hyperstimulation occurred in 9% of women in the misoprostol group compared with 14% in the dinoprostone group (P =.27). The only significant difference in neonatal outcomes was a more frequent presence of thick meconium in the misoprostol group (P =.03). CONCLUSION: We found no difference in terms of effectiveness and safety between low-dose oral misoprostol and vaginal dinoprostone used for induction of labor. This regimen avoids the excessive uterine contractility noted in previous studies, where higher doses of misoprostol were administered at longer intervals. PMID- 12548213 TI - Vascular endothelial growth factor induced functional and morphologic signs of endothelial dysfunction in isolated arteries from normal pregnant women. AB - OBJECTIVE: The purpose of this study was to determine the effects of vascular endothelial growth factor on basal tone, endothelium-dependent dilatation, permeability, and morphologic features of endothelium in isolated arteries from normal pregnant women. We hypothesized that vascular endothelial growth factor might induce signs of endothelial dysfunction. STUDY DESIGN: Arteries (approximately 200 microm) were dissected from subcutaneous fat biopsy specimens that were obtained at cesarean delivery and mounted on a pressure arteriograph. Changes in basal tone, dilatation to bradykinin (1 nmol/L to 3 micromol/L) before, during, and after 3 hours of incubation with vascular endothelial growth factor (0.5 or 1 nmol/L), vascular endothelial growth factor (0.5 nmol/L) plus bosentan (a nonselective endothelin receptor A and B antagonist, 1 micromol/L), or vehicle were compared. Scanning electron microscopy was applied for endothelial morphologic features. Permeability to Evans blue dye was evaluated in arteries after incubation with vascular endothelial growth factor, vascular endothelial growth factor plus angiopoietin-1, or vehicle, and in arteries that were obtained from women with preeclampsia. RESULTS: Basal tone was higher after 60 minutes of incubation with vascular endothelial growth factor (0.5 nmol/L) compared with vehicle (29% +/- 5% [n = 10] vs 10% +/- 4% [n = 7], P =.006). Combination of vascular endothelial growth factor with bosentan failed to increase the tone (n = 4). Bradykinin-mediated dilatation was impaired in arteries that were incubated with vascular endothelial growth factor 0.5 nmol/L (max dilatation: 287% +/- 16% vs 160% +/- 23% [n = 10], P =.0001) or vascular endothelial growth factor 1 nmol/L (max dilatation: 207% +/- 21% vs 88% +/- 4% [n = 3], P =.003). Bradykinin-mediated dilatation was similar after incubation with vehicle (n = 7) or the combination of vascular endothelial growth factor plus bosentan (n = 4). Evans blue dye staining was higher after incubation with vascular endothelial growth factor but was reversed by the addition of angiopoietin-1. Scanning electron microscopy demonstrated the development of intercellular gaps. CONCLUSION: Vascular endothelial growth factor impaired bradykinin-mediated dilatation and enhanced basal tone and permeability. This might indicate a potential role for vascular endothelial growth factor in the development of endothelial dysfunction in pregnancy. Angiopoietin-1 inhibited the vascular endothelial growth factor-induced vascular leakage, which may have therapeutic implications in preeclampsia. PMID- 12548214 TI - Longitudinal serum concentrations of placental growth factor: evidence for abnormal placental angiogenesis in pathologic pregnancies. AB - OBJECTIVE: Complicated pregnancies demonstrate abnormal decidual and placental villous vasculature. We examined maternal concentrations of vascular endothelial growth factor and placental growth factor in normal pregnancies and in pregnancies that were complicated by isolated idiopathic small-for-gestational age (SGA) newborn infants, preeclampsia alone, or preeclampsia with SGA newborn infants at the time of clinical disease and before the onset of clinical signs. STUDY DESIGN: Serum vascular endothelial growth factor and placental growth factor were measured by enzyme-linked immunosorbent assay in cross-sectional and longitudinal cohorts of pregnant nulliparous women. The results were compared by Wilcoxon tests or a mixed-models method, respectively. RESULTS: In the cross sectional study, serum placental growth factor was reduced in abnormal pregnancy relative to control subjects (SGA newborn infants, 18 [P =.04]; preeclampsia, 20; or preeclampsia with small- for-gestational-age newborn infants, 11 [P =.0001]) as early as 15 to 19 weeks of gestation in preeclampsia with SGA newborn infants. Vascular endothelial growth factor was <30 pg/mL in all serum specimens from pregnant women. CONCLUSION: We postulate that decreased placental growth factor production results in abnormalities of placental angiogenesis through direct and indirect effects on other vasculotropic growth factors. PMID- 12548215 TI - Fetal electrocardiography in labor and neonatal outcome: data from the Swedish randomized controlled trial on intrapartum fetal monitoring. AB - OBJECTIVE: Cardiotocography plus automatic ST analysis of the fetal electrocardiography has been shown recently to reduce both the operative delivery rate for fetal distress and the cord artery metabolic acidosis rate. The purpose of this study was to analyze findings that were related to cases with a complicated/adverse neonatal outcome in the Swedish randomized controlled trial. STUDY DESIGN: Of the 4966 term fetuses that were included in the trial, all 351 newborn infants who required special neonatal care were identified. Cases of perinatal death, neonatal encephalopathy, or metabolic acidosis at birth were reviewed. RESULTS: Of the 29 fetuses with adverse/complicated neonatal outcome, 22 fetuses had cardiotocography and ST patterns that indicated a need for intervention, according to the cardiotocography plus ST clinical guidelines. The number of live-born with moderate or severe neonatal encephalopathy showed a significant decrease from 0.33% (8/2447 fetuses) in the cardiotocography-only group to 0.04% (1/2519 fetuses) in the cardiotocography plus ST group. CONCLUSION: Cardiotocography plus ST analysis provides accurate information about intrapartum hypoxia and may prevent intrapartum asphyxia and neonatal encephalopathy by giving a clear alert to the staff members who are in charge. PMID- 12548216 TI - Nucleated red blood cells in polycythemic infants. AB - OBJECTIVE: This study was undertaken to evaluate whether the absolute nucleated red blood cell (RBC) count is elevated in term, appropriate-for-gestational-age (AGA) polycythemic infants. STUDY DESIGN: We compared absolute nucleated RBC counts taken during the first 12 hours of life in term, AGA infants with neonatal polycythemia (n = 29), and in control, nonpolycythemic infants (n = 37). We excluded infants of women with diabetes, hypertension, and alcohol, tobacco, or drug abuse, and those with fetal heart rate abnormalities or low Apgar scores, hemolysis, blood loss, or chromosomal anomalies. RESULTS: There were no differences between groups in birth weight, gestational age, or other demographic or perinatal factors. The hematocrit, RBC count, and absolute nucleated RBC counts were significantly higher and the platelet counts significantly lower in the polycythemic group. Regression analysis that included Apgar scores and gestational age showed a significant correlation of absolute nucleated RBC count with the polycythemia status only (P =.017). CONCLUSION: At birth, term AGA polycythemic infants have increased indices of active erythropoiesis. We speculate that this finding is suggestive of subtle fetal hypoxemia. PMID- 12548217 TI - Vasoactive effects of neurokinin B on human blood vessels. AB - OBJECTIVES: Preeclampsia (PE) is a multisystem disease unique to human pregnancy. Abnormal placentation results in placental hypoperfusion leading to the secretion of a factor(s) by the placenta. Our aim was to investigate whether neurokinin B (NKB) is the circulating factor associated with PE. STUDY DESIGN: Vascular effects of NKB were assessed in blood vessels dissected from myometrial and omental biopsy specimens obtained at caesarean section from normal pregnant women (n = 26) or in mesenteric blood vessels obtained from nonpregnant female Wistar rats (n = 4). RESULTS: Incubation with NKB did not alter endothelial-dependent relaxation of omental or myometrial arteries. NKB produced a dose-dependent relaxation in preconstricted omental arteries and veins. NKB did not affect vasoactive responsiveness of rat mesenteric arteries. CONCLUSION: We conclude from these observations that NKB is not the circulating factor associated with increased vascular resistance in PE. PMID- 12548218 TI - Protective effect of N-acetylcysteine against fetal death and preterm labor induced by maternal inflammation. AB - OBJECTIVE: Intrauterine and maternal systemic infections are proposed causes of preterm labor. The resulting prematurity is associated with 75% of infant mortality and 50% of long-term neurologic handicaps. We hypothesize that free radicals generated in large quantities during an inflammatory response shift the fetomaternal redox balance to an oxidative state, compromising the fetus. Thus, if our working hypothesis is correct, selective inactivation of free radicals with N-acetylcysteine (NAC), an antioxidant and glutathione (GSH) precursor, would improve the outcome of preterm deliveries associated with inflammation. We tested aspects of this hypothesis in an animal model of preterm labor and fetal damage (death). STUDY DESIGN: NAC (1 g/kg) was administered orally to C57Bl/6 mice injected intraperitoneally with either 10 microg lipopolysaccharide (LPS) or saline solution (CRL) on day 16 of gestation. The latency period (time from injection to delivery of the first pup) and fetal viability were recorded. To discriminate between an effect of prematurity from an effect of inflammation, and to document any improvement in survival, mice were killed at 3, 6, and 16 hours after injection. Maternal and fetal redox states were approximated by measuring hepatic GSH. RESULTS: Each C57Bl/6 LPS-treated mouse delivered prematurely after a significantly shorter latency period (LPS: 16.8 hours [95% CI 15.9-17.6] vs CRL: 54.7 hours [95% CI 43.8-65.5]). NAC doubled the latency interval of LPS treated animals to 35.2 hours (95% CI 21.0-49.2). LPS alone resulted in a 100% rate of stillbirth. Fifty-eight percent of fetuses were already dead 16 hours after LPS. In contrast, only 33% of fetuses were dead 16 hours after LPS (P =.001) when NAC was given. LPS was followed by a reduction in maternal (LPS: 26.3 nmol/mg [95% CI 19.9-32.8] vs CRL: 41.3 nmol/mg [95% CI 34.7-47.9, P <.01]) and fetal GSH (LPS: 19.7 nmol/mg [95% CI 11.7-27.8] vs CRL: 34.5 nmol/mg [95% CI 32.0 37.0, P <.001]). This decline was reversed by NAC (NAC/LPS maternal GSH: 37.0 nmol/mg [95% CI 22.5-51.5] and fetal GSH: 28.4 nmol/mg [95% CI 22.8-33.9]). Importantly, maternal liver GSH impacted on fetal survival. NAC/LPS mothers with living pups 16 hours after LPS had significantly higher liver GSH compared with NAC/LPS mothers whose pups died in utero. In fact, all NAC-treated mice whose hepatic GSH exceeded 20 nmol/mg had living fetuses at 16 hours. CONCLUSION: Maternal inflammation in C57Bl/6 mice results in oxidative stress associated with maternal and fetal GSH depletion. Oxidative stress damages the fetus independent of prematurity. Restoration of maternal and fetal oxidative balance by NAC protects the fetus and reduces the rate of preterm birth. PMID- 12548219 TI - Preterm premature rupture of membranes: risks of recurrent complications in the next pregnancy among a population-based sample of gravid women. AB - OBJECTIVE: The purpose of this study was to ascertain the predictive value of antecedent preterm premature rupture of membranes for recurrent preterm premature rupture of membranes and preterm delivery rates in the next pregnancy compared with background rates among a population-based sample of women at a single institution. STUDY DESIGN: Records of patients with index singleton pregnancies that were complicated by preterm premature rupture of membranes whose next delivery resulted in a delivery at >or=20 weeks at the same institution were reviewed for the incidence and gestational age of recurrent preterm premature rupture of membranes and preterm delivery. All subjects were patients of physicians whose obstetric practices were based at a single institution. Background rates of preterm premature rupture of membranes and preterm delivery in this population were generated from a systematically selected comparison group composed of the two deliveries after each of the study group's second delivery. RESULTS: The rates of recurrent preterm premature rupture of membranes (16.7%) and preterm delivery (34.2%) in the 114 study group patients were substantially greater (odds ratio, 20.6; 95% CI, 4.7-90.2; and odds ratio, 3.6; 95% CI, 2.1 6.4) than noted background rates (0.96% and 12.5%) but considerably less than the recurrence rates of either preterm premature rupture of membranes or preterm delivery that were reported by others. The gestational age of preterm premature rupture of membranes in the index pregnancy affected neither the magnitude of risk nor the gestational age of recurrent preterm premature rupture of membranes or preterm delivery in the subsequent pregnancy. Stratification of outcome measures into three subgroups that were based on the gestational age of index preterm premature rupture of membranes demonstrated no significant differences in the incidence of preterm premature rupture of membranes or preterm delivery. CONCLUSION: After a pregnancy that was complicated by preterm premature rupture of membranes, the risk for recurrent preterm premature rupture of membranes is increased by 20-fold and for recurrent preterm delivery by almost 4-fold. Gestational age of antecedent preterm premature rupture of membranes is predictive of neither risk nor timing of recurrent complications. Estimates of recurrence risks appear to be moderated by limiting analysis to a population based sample of gravid women when compared with previous studies. PMID- 12548220 TI - A comparison between middle cerebral artery peak systolic velocity and amniotic fluid optical density at 450 nm in the prediction of fetal anemia. AB - OBJECTIVE: The purpose of this study was to compare fetal middle cerebral artery peak systolic velocity with amniotic fluid delta optical density at 450 nm in the prediction of fetal anemia. STUDY DESIGN: A prospective study that involved 28 singleton pregnancies that were at-risk for fetal anemia was carried out in a tertiary teaching hospital. Middle cerebral artery peak systolic velocity was measured immediately before the determination of deltaoptical density at 450 nm and fetal hemoglobin concentration. Sensitivities and predictive values for fetal anemia were examined. RESULTS: Fetal hemoglobin concentrations correlated significantly with middle cerebral artery peak systolic velocity (correlation coefficient, -0.77; P <.0001) and deltaoptical density at 450 nm zones (correlation coefficient, -0.56; P =.0025). Middle cerebral artery peak systolic velocity was >1.5 multiples of the median in 15 of 28 cases (54%); for this cutoff value, the sensitivity and positive-predictive values for a hemoglobin deficit of >-3SD were 75% and 60% and for a hemoglobin deficit of >-5SD were 100% and 47%, respectively. The corresponding values for deltaoptical density at 450 nm zone III (6/28 cases, 21%) were 0% (hemoglobin deficit, <-3SD) and 86% and 100% (hemoglobin deficit, <-5SD). CONCLUSION: Middle cerebral artery peak systolic velocity and amniotic fluid optical density at 450 nm are both useful in the prediction of fetal anemia. However, Doppler examination has the advantage of being a noninvasive method that can help reduce the number of invasive procedures in pregnancies that are at-risk for fetal anemia. PMID- 12548221 TI - Cesarean delivery for twins: a systematic review and meta-analysis. AB - OBJECTIVE: We undertook a systematic review and meta-analysis to determine whether a policy of planned cesarean section or vaginal delivery is better for twins. STUDY DESIGN: We searched MEDLINE and EMBASE from 1980 through May 2001 using combinations of the following terms: twin, delivery, cesarean section, vaginal birth, birth weight, and gestational age. Studies that compared planned cesarean section to planned vaginal birth for babies weighing at least 1500 g or reaching at least 32 weeks' gestation were included. We computed pooled odds ratios for perinatal or neonatal mortality, low 5-minute Apgar score, neonatal morbidity, and maternal morbidity. The infant was the unit of statistical analysis. Results were considered statistically significant if the 95% CI did not encompass 1.0. RESULTS: We retrieved 67 articles, 63 of which were excluded. Four studies with a total of 1932 infants were included in the analysis. A low 5 minute Apgar score occurred less frequently in twins delivered by planned cesarean section (odds ratio, 0.47; 95% CI, 0.26-0.88) principally because of a reduction among twins if twin A was in breech position (odds ratio, 0.33; 95% CI, 0.17-0.65). Twins delivered by planned cesarean section spent significantly longer in the hospital (mean difference, 4.01 days; 95% CI, 0.73-7.28 days). There were no significant differences in perinatal or neonatal mortality, neonatal morbidity, or maternal morbidity. CONCLUSION: Planned cesarean section may decrease the risk of a low 5-minute Apgar score, particularly if twin A is breech. Otherwise, there is no evidence to support planned cesarean section for twins. PMID- 12548222 TI - Antenatal prediction of pulmonary hypoplasia by acceleration time/ejection time ratio of fetal pulmonary arteries by Doppler blood flow velocimetry. AB - OBJECTIVE: The purpose of this study was to develop a new method for the antenatal prediction of pulmonary hypoplasia by Doppler blood flow velocimetry. STUDY DESIGN: One hundred seventy-seven fetuses (160 normal fetuses and 17 fetuses with congenital anomalies that may affect fetal lung growth and/or development) were studied. Blood flow waveforms at the main branches of the pulmonary arteries were recorded by Doppler echocardiography from 20 to 39 weeks of gestation. The ratio of acceleration time to ejection time was calculated from the waveform as a parameter to predict pulmonary hypoplasia. RESULTS: Doppler waveform of normal fetal pulmonary artery showed a "spike-and-dome" pattern. The normal values of acceleration time/ejection time ratio from the right and left pulmonary arteries were 0.17 +/- 0.04 and 0.15 +/- 0.04, respectively. These values were not significantly altered through the gestational age observed in this study. The acceleration time/ejection time ratio of either right or left pulmonary artery was measured successfully in all cases of fetal congenital anomalies. In 8 of 17 fetuses, acceleration time/ejection time ratio was measured at both of the pulmonary arteries. Because of a congenital anomaly that affected the fetal lung or thorax asymmetrically (as in congenital diaphragmatic hernia or congenital cystic adenomatoid malformations of the lung), the acceleration time/ejection time ratio of both of the pulmonary arteries could be measured in only 5 of 13 fetuses. The technical difficulties for the measurement always existed in the affected side. Eleven of the 17 fetuses with congenital anomalies survived without signs of clinical pulmonary hypoplasia or persistent pulmonary hypertension of the newborn infant. The fetuses revealed normal acceleration time/ejection time ratio from at least one pulmonary artery. The remaining 6 fetuses died of pulmonary hypoplasia, and the diagnosis was confirmed by autopsy or clinical findings. Of those 6 fetuses, 5 fetuses demonstrated the acceleration time/ejection time ratio below normal in one side, and the ratio could not be obtained on the other side; 1 fetus showed the acceleration time/ejection time ratio below the normal range in both sides. CONCLUSION: The acceleration time/ejection time ratio by Doppler velocimetry that was obtained at the main branches of fetal pulmonary artery was consistent throughout gestational age from 20 to 39 weeks. This ratio appears to be an accurate parameter with which to predict the subsequent development of pulmonary hypoplasia and clinical outcomes of the newborn infants with high positive and negative predictive values (positive predictive value, 100%; negative predictive value, 100%). PMID- 12548223 TI - Relaxin causes proliferation of human amniotic epithelium by stimulation of insulin-like growth factor-II. AB - OBJECTIVE: The study was conducted to determine whether relaxin has a proliferative effect on amniotic epithelial cells and to show that this effect is caused by its stimulation of the insulin-like growth factor-II (IGF-II) gene. STUDY DESIGN: Immunolocalization and Northern analysis were used to confirm the expression of IGF-II by the fetal cells in the membranes. Human amniotic epithelial (WISH) cells were treated with doses of IGF-II or human relaxin and their proliferative effects measured. The mechanism of the effect of relaxin on cellular proliferation was studied with the use of an IGF-II-blocking antibody and Northern analysis for IGF-II gene expression after treatment with relaxin. An in vivo correlate was sought by quantitation of relaxin gene expression in 10 fetal membranes from women with normally grown and large for gestational age infants. RESULTS: The amniotic epithelial and cytotrophoblast cells of the fetal membranes expressed IGF-II, as did the amniotic epithelial-like (WISH) cell line. Treatment of WISH cells with IGF-II or relaxin caused a significant (P <.03) and dose-related increase in WISH cell proliferation over 5 days. The concurrent treatment with a blocking antibody to IGF-II significantly decreased the proliferative response to IGF-II (P <.002) and relaxin (P <.002). Treatment with relaxin caused a significant increase (P <.003) in the transcription of IGF-II in 24 hours. In fetal membranes, the levels of relaxin gene expression correlated with fetal membrane surface area (r = 0.76) and was significantly greater (P <.008) in the membranes from macrosomic infants (4020-4729 g) compared with those normally grown (2855-3830 g). CONCLUSION: IGF-II and relaxin both caused the proliferation of WISH cells. Concurrent treatment with an IGF-II-blocking antibody abrogated the proliferative effects of both hormones. Relaxin increased the transcription of IGF-II, and its expression levels in the fetal membranes correlated with the membrane surface area as well as neonatal birth weight. These data suggest that relaxin is a growth factor for the fetal membranes. PMID- 12548224 TI - Results of systematic screening for minor degrees of fetal renal pelvis dilatation in an unselected population. AB - OBJECTIVE: The purpose of this study was to determine the incidence of minor degrees of renal pelvis dilatation that is detected by antenatal ultrasound scanning in an unselected population and its value in the prediction of significant uropathies. STUDY DESIGN: This prospective study was conducted over a 24-month period. Infants with an anteroposterior pelvic diameter of >or=4 mm in the second trimester and/or >or=7 mm but <15 mm in the third trimester were enrolled. RESULTS: Pyelectasis was found in 4.5% of 5643 fetuses (1.5% with significant uropathy). Among the 213 infants whose cases were followed, 132 infants (62%) had renal anomalies, but only 83 infants (39%) had significant uropathies. The ability of the third-trimester renal pelvis dilatation to predict renal abnormalities showed a positive predictive value of 69%. Pyelectasis that was detected only in the second trimester revealed a significant uropathy in 12% of the infants. CONCLUSION: Pyelectasis was found in 4.5% of fetuses. The third trimester anteroposterior renal pelvis diameter of >or=7 mm was the best ultrasound criterion to predict postnatal uropathies. PMID- 12548225 TI - Increase in maternal placental growth hormone during pregnancy and disappearance during parturition in normal and growth hormone-deficient pregnancies. AB - OBJECTIVE: The purpose of this study was to evaluate placental growth hormone levels in maternal circulation throughout pregnancy in normal and growth hormone deficient women with the use of a specific assay and to determine the clearance of placental growth hormone from maternal circulation after birth. STUDY DESIGN: Seventeen healthy pregnant women and 1 patient with growth hormone deficiency substituted with recombinant growth hormone during pregnancy participated in a longitudinal study from early pregnancy until birth with repetitive blood sampling and measurement of placental growth hormone levels throughout pregnancy. Furthermore, serial blood samples were drawn before, during, and after elective caesarean deliveries in 5 healthy women to calculate the half-life of placental growth hormone. Placental growth hormone was measured with the use of two monoclonal antibodies in a commercially available solid-phase iodine 125-labeled immunoradiometric assay (Biocode, Liege, Belgium). RESULTS: Placental growth hormone levels were detectable from as early as 8 weeks of gestation in some of the women and increased throughout gestation, with a maximum at approximately 35 to 36 weeks of gestation (13.7 ng/mL; range, 5.9-24.4 ng/mL) and large interindividual variations. Placental growth hormone levels did not correlate with birth weight or placental weight. In the patient with isolated growth hormone deficiency, placental growth hormone levels were detectable from 11 weeks of gestation (3.4 ng/mL) and increased throughout pregnancy to 13.9 ng/mL, which is similar to values that are obtained in the healthy pregnant women. Substitution therapy with recombinant human growth hormone did not suppress the increase in placental growth hormone. We found a mean half-life of placental growth hormone of 13.8 minutes (range, 11.5-15.2 minutes) in healthy pregnant women and an apparently similar half-life of placental growth hormone (15.8 minutes) in the growth hormone-deficient patient, assuming a monoexponential disappearance of placental growth hormone during the first 30 minutes after the delivery. After the initial 30 minutes, approximately 75% (range, 65%-89%) of the placental growth hormone had been cleared from the maternal circulation. CONCLUSION: Levels of placental growth hormone in maternal circulation increase throughout pregnancy from as early as 8 weeks of pregnancy, with maximum levels around the week 35 of gestation. The pregnancy-induced rise in placental growth hormone levels in the growth hormone-deficient patient was comparable to the rise seen during normal pregnancies and was not suppressed by the concurrent human growth hormone treatment. We speculate that maternal serum levels of placental growth hormone reflect placental function and fetal growth. However, further studies are needed to evaluate the potential clinical use of placental growth hormone determinations. PMID- 12548226 TI - Dexamethasone or interleukin-10 blocks interleukin-1beta-induced uterine contractions in pregnant rhesus monkeys. AB - OBJECTIVE: The purpose of this study was to determine whether treatment with the immune modulators dexamethasone or interleukin-10 prevents interleukin-1beta induced uterine contractions in a nonhuman primate model. STUDY DESIGN: Thirteen chronically instrumented rhesus monkeys at 135 +/- 1 days of gestation (term, 167 days) received one of three interventions: (1) intra-amniotic interleukin-1beta (10 microg) infusion with maternal dexamethasone (1 mg/kg) intravenously every 6 hours for 1 day before interleukin-1beta and for 2 days thereafter (n = 4), (2) intra-amniotic interleukin-1beta infusion with maternal interleukin-10 (25 microg/kg) given intravenously and 100 microg interleukin-10 given intra amniotically before the interleukin-1beta and continued every 8 hours for 3 days (n = 5), and (3) intra-amniotic interleukin-1beta administered alone (n = 5). Uterine activity was monitored continuously and quantified as the hourly contraction area (millimeters of mercury times seconds per hour) in all groups until delivery. Amniotic fluid was sampled for leukocyte counts and assayed for prostaglandins E(2) and F(2)alpha, cytokines interleukin-1beta, interleukin-6, interleukin-8, tumor necrosis factor-alpha, interleukin-10, and interleukin-1 receptor antagonist by specific assays. Maternal and fetal blood were assayed for cortisol, dehydroepiandrosterone sulfate, and estradiol. RESULTS: Interleukin 1beta infusion in the absence of immune modulators resulted in an increase in uterine activity and amniotic fluid proinflammatory cytokines, prostaglandins, and leukocytes. Dexamethasone and interleukin-10 treatment significantly reduced interleukin-1beta-induced uterine contractility (P <.05) and amniotic fluid prostaglandins (P <.05) but not interleukin-8 or interleukin-1 receptor antagonist. Amniotic fluid interleukin-6 and maternal and fetal cortisol, dehydroepiandrosterone sulfate, and estradiol concentrations were reduced by dexamethasone (P <.05), whereas tumor necrosis factor-alpha levels and leukocyte counts were attenuated by interleukin-10 treatment (P <.05). An inverse relationship was noted between amniotic fluid interleukin-10 concentrations and interleukin-1beta-induced uterine activity (r = -0.74, P <.05). CONCLUSION: Dexamethasone and interleukin-10 exert similar inhibitory effects on interleukin 1beta-induced uterine activity, which appears to be mediated by a decrease in prostaglandin production. Reduced estrogen biosynthesis or suppression of tumor necrosis factor-alpha and leukocyte migration may contribute to the tocolytic actions of dexamethasone and interleukin-10, respectively. Dexamethasone and interleukin-10 are likely to be useful adjuncts in the treatment of preterm labor that is associated with inflammation or infection. PMID- 12548227 TI - Perinatal outcomes in preeclampsia that is complicated by massive proteinuria. AB - OBJECTIVE: Current treatment of preeclampsia no longer mandates delivery for proteinuria of >5 g per 24 hours. We sought to determine whether delayed delivery of preeclampsia with massive proteinuria (>10 g/24 h) increased maternal or neonatal morbidity. STUDY DESIGN: Records of all women with preeclampsia who were delivered at <37 weeks of gestation between January 1, 1997, and June 30, 2001, were reviewed. Patients with underlying renal disease or multiple gestation were excluded. Patients were characterized as having mild (<5 g/24 h), severe (5-9.9 g/24 h), or massive (>10 g/24 h) proteinuria. Outcomes were compared using the chi(2) test, one-way analysis of variance, or Fisher exact test. RESULTS: Two hundred nine patients met the inclusion criteria: 125 patients had mild proteinuria, 43 patients had severe proteinuria, and 41 patients had massive proteinuria. No significant differences in maternal morbidity were seen. Massive proteinuria was associated with earlier onset of preeclampsia, earlier gestational age at delivery, and higher rates of prematurity complications. After correction for prematurity, massive proteinuria has no significant effect on neonatal outcomes. CONCLUSION: Women with preeclampsia and massive proteinuria did not have increased maternal morbidity compared with women with severe or mild proteinuria. Massive proteinuria appears to be a marker for early-onset disease and progression to severe preeclampsia. Neonatal morbidity appears to be a function of prematurity rather than of massive proteinuria itself. PMID- 12548228 TI - Intrapartum fever at term: serum and histologic markers of inflammation. AB - OBJECTIVE: This study was undertaken to determine whether intrapartum fevers at term are associated with markers of acute inflammation in maternal, fetal, and placental compartments. STUDY DESIGN: Term cases with intrapartum fever (temperature >/=100.4 degrees F) were recruited with gestational age-matched controls. Maternal serum and umbilical vein blood were collected and assayed for interleukin-6 (IL-6) levels. Placentas were examined for histologic chorioamnionitis. Demographic and clinical data were collected and compared between cases and controls. RESULTS: Forty-seven case-control pairs were analyzed. Maternal IL-6 levels were higher in cases than in controls (median of 145 pg/mL vs 42 pg/mL, P <.0001). Umbilical vein IL-6 levels also were higher in cases than controls (median 9 pg/mL vs 3.5 pg/mL, P =.01), but more than half of levels in cases were below 11 pg/mL. Only 31.1% of febrile cases had moderate or severe histologic chorioamnionitis. Multivariable logistic regression identified maternal serum IL-6 levels, nulliparity, and number of vaginal examinations as the major predictors of intrapartum fever at term. CONCLUSION: The maternal inflammatory response as measured by maternal serum IL-6 levels is a strong marker for term intrapartum fever. The much weaker association of fetal and placental inflammatory responses suggest a smaller than expected contribution of intra-amniotic inflammation to term intrapartum fevers. PMID- 12548229 TI - Placenta previa in singleton and twin births in the United States, 1989 through 1998: a comparison of risk factor profiles and associated conditions. AB - OBJECTIVE: The purpose of this study was to compare risk factor profiles for placenta previa between singleton and twin live births. STUDY DESIGN: This cohort study was based on United States natality data files (1989 through 1998) and comprised 37,956,020 singleton births and 961,578 twin births. Women who were diagnosed with placenta previa were included only if they were delivered by cesarean delivery. Risk factors for placenta previa that were examined included sociodemographic (age, gravidity, education, marital status, and race), behavioral (prenatal care, smoking, and alcohol use), previous preterm birth, and medical and obstetric factors. Effect modification between maternal age and gravidity and the dose-response relationship with number of cigarettes smoked/day on placenta previa risk were also evaluated. RESULTS: The rate of placenta previa was 40% higher among twin births (3.9 per 1,000 live births, n = 3,793 births) than among singleton births (2.8 per 1,000 live births, n = 104,754 births). Comparison of risk factors for placenta previa between the singleton and twin births revealed fairly similar risk factor profiles. Compared with primigravid women <20 years old, the risk for placenta previa increased by advancing age and by increasing number of pregnancies among both singleton and twin births. The number of cigarettes smoked per day also showed a dose-response trend for placenta previa risk in the two groups. CONCLUSION: The increased rate of placenta previa among twin births underscores the need to monitor carefully such pregnancies with heightened suspicion and awareness for the development of this condition. PMID- 12548230 TI - Pregnancy outcome after gestational exposure to mebendazole: a prospective controlled cohort study. AB - OBJECTIVE: Mebendazole is an anthelmintic that is commonly needed in women of reproductive age. Its use in pregnancy is a reason for concern for women and their health care providers. The purpose of this study was to examine the fetal safety of mebendazole. STUDY DESIGN: The Israeli Teratogen Information Service prospectively collected and followed 192 pregnancies exposed to mebendazole in pregnancy, 71.5% of whom had first-trimester exposure. Pregnancy outcome was compared with that of a matched control group, who were counseled for nonteratogenic exposure. RESULTS: There was no increase in the rate of major malformations between the groups (5/150 pregnancies [3.3%; mebendazole] vs 3/175 pregnancies [1.7%; nonteratogenic control subjects]; P =.478). There was a higher rate of elective terminations of pregnancy in the exposed group compared with the control group (22/192 pregnancies [11.5%; mebendazole] vs 3/192 pregnancies [1.6% [nonteratogenic control subjects]; P =.000). CONCLUSION: This study suggests that mebendazole does not represent a major teratogenic risk in humans when it is used in the doses that are used commonly for pinworm (Enterobius vermicularis) infestation. PMID- 12548231 TI - Female sexual dysfunction in postmenopausal women: systematic review of placebo controlled trials. AB - OBJECTIVE: This systematic review includes all randomized and placebo-controlled trials (RCTs) of treatment for female sexual dysfunction (FSD) in postmenopausal women published since 1990. STUDY DESIGN: Electronic database and manual bibliography searches were conducted to identify all relevant publications. RESULTS: Only six RCTs have been done to assess the effects of different therapies on sexual function in postmenopausal women: one with sildenafil citrate (Viagra), three with hormone replacement therapy, and two with tibolone. CONCLUSIONS: In women with FSD, many treatments that are used in practice are not supported by adequate evidence. Although an improvement of sexual function was reported with tibolone and the combination of estrogen-androgen therapy, it still remains unclear which groups of postmenopausal women with FSD would benefit most from these therapies. The adverse effects of testosterone replacement therapy should be assessed against the effects of placebo in RCTs with larger sample sizes and longer duration. PMID- 12548232 TI - Association between use of antenatal magnesium sulfate in preterm labor and adverse health outcomes in infants. PMID- 12548234 TI - Association between use of antenatal magnesium sulfate in preterm labor and adverse health outcomes in infants. PMID- 12548235 TI - The impact of a single- or double-layer closure on uterine rupture. PMID- 12548237 TI - Are users of alcohol in pregnancy necessarily alcohol abusers? PMID- 12548239 TI - The use of misoprostol as a cervical ripening agent in operative hysteroscopy. PMID- 12548241 TI - Physician attitudes concerning legal coercion of pregnant alcohol and drug users. PMID- 12548243 TI - Direct visualization of the uterine cavity provides a better triage than saline hysterography. PMID- 12548245 TI - Procedural risks versus theology: chorionic villus sampling for Orthodox Jews at less than 8 weeks' gestation. PMID- 12548246 TI - Procedural risks versus theology: chorionic villus sampling for Orthodox Jews at less than 8 weeks' gestation. PMID- 12548248 TI - Pelvic organ support in nulliparous pregnant and nonpregnant women. PMID- 12548250 TI - Which antimicrobial impregnated central venous catheter should we use? Modeling the costs and outcomes of antimicrobial catheter use. AB - BACKGROUND: Catheter-related bloodstream infections are costly and associated with substantial morbidity and mortality. Trials suggest that central venous catheters impregnated with minocycline/rifampin, although more expensive, are clinically superior to chlorhexidine/silver sulfadiazine impregnated catheters. It remains unclear whether minocycline/rifampin catheters are cost-effective for all high-risk patients or only those requiring longer-term catheterization. METHODS: We developed a series of decision models with patient-level clinical trial data to determine whether minocycline/rifampin catheters are cost-effective for patients requiring various durations of catheterization. We calculated incremental cost-effectiveness ratios for patients catheterized for durations ranging from 1 to 25 days. RESULTS: The data were too sparse to estimate cost effectiveness for patients catheterized less than 8 days. The probability that minocycline/rifampin catheters were cost-effective compared with chlorhexidine/silver sulfadiazine catheters in patients catheterized for 8 days was 91%. The probability that the minocycline/rifampin catheters in patients catheterized 13 days or longer resulted in cost savings was more than 95%. CONCLUSIONS: Our analysis suggests that central venous catheters coated with minocycline/rifampin are cost-effective for patients catheterized for at least 1 week and lead to overall cost savings when patients are catheterized for 2 weeks or longer. Policies for the use of antimicrobial catheters in high-risk patients should reflect patients' expected duration of catheterization. PMID- 12548251 TI - Incidence of urinary tract infections in patients with short-term indwelling urethral catheters: a comparison between a 3-day urinary drainage bag change and no change regimens. AB - BACKGROUND: The current practice of caring for hospitalized patients with indwelling urethral catheters in Siriraj Hospital is to change the drainage bag every 3 days. In an extensive medical literature search, no evidence was noted to support this practice. OBJECTIVE: The purpose of this study was to compare the incidence of catheter-associated urinary tract infections (UTI) in hospitalized patients with indwelling catheters who receive a drainage bag change every 3 days with the incidence of UTI in patients who receive no bag change. DESIGN: This study was a randomized controlled trial. Participants and study procedures: Of the patients at Siriraj Hospital, 153 with an indwelling urinary catheter for at least 3 days were randomized to a 3-day drainage bag change or a no change regimen. A urine sample was obtained from each patient for culture every 7 days, on the day the catheter was removed, or the day the patient was suspected of having a UTI. RESULTS: Of the 153 study patients, 79 were randomized to the 3-day bag change regimen, and 74 patients were in the no-change group. Both groups were comparable for all baseline characteristics. The incidence of symptomatic UTI was 13.9% in the 3-day drainage bag change group and 10.8% in the no change group (P =.7). The incidence of asymptomatic UTI was 36.7% in the 3-day bag change group and 36.5% in the no change group (P =.9). CONCLUSION: There is no evidence for the necessity of a bag change every 3 days at Siriraj Hospital; the urine bag can be left longer than 3 days. However, the appropriate frequency of urinary drainage bag change needs additional study because the sample size in this study does not rule out a false-negative result. PMID- 12548252 TI - Investigation of single-use versus reusable infectious waste containers as potential sources of microbial contamination. AB - BACKGROUND: Laws require that infectious waste be segregated from noninfectious waste. Companies certified to dispose of infectious waste offer both reusable and single-use containers. The focus of this study was to determine if there would be a microbiologic advantage to the use of one type of container over another in a burn hospital. METHODS: Monthly swab cultures were taken from the tops of >250 infectious waste containers during 2 years. Bacteria and fungi were identified. In a substudy swab cultures were taken from an area of reusable tops before and after cleaning to evaluate the efficacy of cleaning on both the number and type of microbes present. Infection rates for acute patients were compared before and after control measures were instituted to decrease microbial transfer from infectious waste containers to patients. RESULTS: Cultures taken from reusable boxes when received from the container company showed that >99% were contaminated with bacteria or fungi; most were normal environmental or skin flora, but some cultures showed microorganisms that can be potentially harmful to patients with compromised immunity. Wiping the lids with a phenolic disinfectant decreased both the total microbial load (P <.001) and the variety of microbes present (P <.001). In contrast, only 10% of the incoming single-use boxes showed any contamination. Infection rates dropped from 5.8 to 3.2 per 100 burn patients (P <.05) after the institution of cleaning and other changes made to decrease the possibility of microbial transfer from the infectious waste boxes to the patients. CONCLUSIONS: Upon delivery, significantly fewer single-use infectious waste boxes were contaminated than reusable ones (P <.001). Extra infection control measures were needed when reusable infectious waste boxes were used in areas housing patients with compromised immunity. Facilities need be aware of the possible contamination of reusable infectious waste containers with microorganisms capable of causing nosocomial infections in patients who are compromised. PMID- 12548253 TI - Trends in antibiotic use and cost and influence of case-mix and infection rate on antibiotic-prescribing in a long-term care facility. AB - BACKGROUND: Antibiotic use and cost indicators have been developed specifically for the long-term care facility (LTCF) setting. Approximately 50% of the variation in these indicators was explained by the variation in infection rate. The objectives of this study were to further assess the utility of the antibiotic use and cost indicators in a different LTCF and to determine the correlation of case-mix and these indicators. METHODS: Antibiotic use and cost indicators were collected monthly by unit (N = 10) and by physician (N = 6) at a 433-bed LTCF in Syracuse, New York, from February 1999 to September 2001. Indicators included incidence (number of antibiotic courses per 1000 resident care-days) of antibiotic use, antibiotic utilization ratio ([AUR]; ratio of the number of antibiotic-days to the number of resident care-days), cost per antibiotic-day, and cost per resident care-day. Case-mix variation was measured with the case-mix index (CMI) of the Resource Utilization Group II system. Simple linear and multilinear regression analyses were used to evaluate correlations of continuous variables. RESULTS: Among the 10 units or 6 physicians, there was a significant difference in the average values for all indicators. Correlation between unit- or physician-specific CMI and antibiotic use or cost indicators or infection rate was poor. However, there was a significant positive correlation between unit- or physician-specific infection rate and incidence of antibiotic use, AUR, and cost per resident care-day but not cost per antibiotic-day. With use of multilinear regression analysis to control for CMI and cost per antibiotic-day, infection rate was a significant predictor of incidence of antibiotic use (R2 = 0.65; P <.001) and AUR (R2 = 0.78; P <.001). CONCLUSIONS: This study provides further evidence that the antibiotic use and cost indicators developed specifically for LTCFs can detect significant variation among units within a facility and among physicians. However, there was no correlation between CMI and antibiotic use or cost indicators. After controlling for case-mix variation and cost per antibiotic day, variation in infection rate explained most of the variation in incidence of antibiotic use and AUR. PMID- 12548254 TI - Impact of dedicated space, dialysis equipment, and nursing staff on the transmission of hepatitis C virus in a hemodialysis unit of the middle east. AB - BACKGROUND: Infection with the hepatitis C virus (HCV) is endemic in hemodialysis (HD) units, especially in Middle Eastern countries. The meticulous isolation policy recommended for patients with the hepatitis B virus (HBV) in an HD unit resulted in a significant drop in HBV incidence globally. This study was developed to prospectively investigate the impact of an identical isolation policy on incidence of nosocomial HCV infection in this HD unit of the Middle East. METHODS: In phase I of the study, we retrospectively reviewed the records of 189 patients with a mean age of 47.5 +/- 11.4 years (range, 15-85 years) who were receiving maintenance HD from December 7, 1995, to December 6, 2000, for the mean duration of 73 +/- 6.3 months (range, 3-144 months) to record the prevalence of HCV. Factors such as blood transfusions and dialytic age (time span that patient has received dialysis since its initiation) implicated in transmission of HCV in the HD unit also were recorded. Phase II involved stringent isolation of anti-HCV positive patients detected during phase I through provision of dedicated space, dialysis equipment, and nursing staff from December 7, 2000, to December 6, 2001. Liver function and anti-HCV tests were repeated for all the 198 patients every 6 months to identify new HCV seroconversions. RESULTS: An HCV prevalence rate of 43.9% (83/189) and an annual HCV seroconversion rate of 6.8% were identified in this cohort. No significant association with blood tranfusion was observed. Eighty-three anti-HCV positive (43.9%) patients had a mean dialytic age of 48.5 +/- 14.2 months compared with 25.0 +/- 8.6 months among 106 (56.1%) anti HCV negative patients (relative risk [RR], 1.89; 95% confidence interval [CI], 1.39-5.86; P <.001). Only 2 new HCV seroconversions (1.01% [2/198]) were identified. CONCLUSIONS: Evidently, the sharing of facilities in a high-risk HD environment for a prolonged dialytic age facilitates the nosocomial transmission of HCV infection. A significant decline of annual seroconversion rate from 6.8% to 1.01% (odds ratio [OR], 7.535; 95% CI, 1.598-48.89; P <.005) suggests that a comprehensive, strictly enforced isolation policy for HCV-positive patients may play a significant role in limiting HCV transmission in HD units, just as it has in drastically reducing HBV transmission in these settings. PMID- 12548255 TI - Evaluation of infectious complications of the implantable venous access system in a general oncologic population. AB - OBJECTIVES: This study was conducted to evaluate the infectious morbidity associated with 2 common types of implantable port systems used in a cancer center in Taiwan. METHODS: This study is retrospective and descriptive. Five hundred seventy-two patients who received implantable ports at the cancer center between January 1994 and December 1998 were included. The following 4 types of catheter-related infections were defined in this study: bloodstream, pocket, exit site, and tunnel. Semiquantitative methods (rolling plate) were used for cultures of removed catheters. RESULTS: The average duration for ports after placement in patients was 358 days (range, 1-1742 days), and the median duration was 242 days. The port-related infection rate was 5.6% in 586 ports and 0.15 per 1000 device days. Our data indicate that when a port is in use, more frequent dressing changes of the needle exit site (every 3 days vs every 7 days) appear to have no significant impact on the catheter-related infection rate (5.2% vs 7.3%, P =.3). Our observation also indicates that if the catheter is used for total parenteral nutrition (TPN), the patient has a significantly increased chance of developing an infection from the Candida species (71% vs 8%, P =.005). Furthermore, the catheters associated with infections caused by the Candida species had a shorter catheter life (median, 44 days). CONCLUSIONS: Infection is the most common complication of venous port system use among cancer patients. According to our data, it is safe and cost-effective to change exit-site needles and transparent dressings every 7 days. It is clearly demonstrated by our study that patients with ports used for the administration of TPN had a significantly greater chance of developing infection from the Candida species. Further investigation is needed to compare the relative advantages of using ports versus other devices for the administration of TPN in cancer patients. PMID- 12548256 TI - Evaluation of aloe vera gel gloves in the treatment of dry skin associated with occupational exposure. AB - OBJECTIVE: An examination glove that delivers aloe vera (AV) gel to the gloved hand was studied in 30 adult females with bilateral occupational dry skin with or without irritant contact dermatitis (with or without erythema, fissures, and excoriations). METHODS: All participants were factory assembly-line workers with repeated superficial skin trauma who attributed their dry, irritated, emollient dependent skin to a common cause (occupational exposure). Participants were sequentially enrolled (after written informed consent, n = 29 evaluable participants) into an open, contralateral comparison study to evaluate efficacy of AV glove use 8 h/day to one hand versus no use to the opposite hand for 30 days, followed by 30 days rest, followed by 10 days of repeated use. Participant's dorsal hands were documented by standardized photos at baseline, during, and at the end of study. RESULTS: Unblinded investigator baseline assessment rated dry skin as mild to moderate (n = 27), or moderate to severe (n = 2). Mean time to noticeable improvement for the AV glove hand was 3.5 days (range: 2-6 days) whereas marked improvement was 10.4 days (range: 7-17 days) for the AV glove hand. No improvement was detected for nonglove hands. Blinded photo assessment was rated independently by dermatology research staff. End-of-study mean global assessment of AV glove hands versus nonglove hands was 1.3 for AV glove hand (0 = no change, 1 = good [10%-89% global improvement], 2 = marked improvement [90%-100% global improvement]) versus 0 for nonglove hand (P <.0001). Mean global end-of-study assessments by the participants = 2.0 for AV glove hand versus 0 for nonglove hand. CONCLUSION: Dry-coated AV gloves that provide for gradual delivery of AV gel to skin produced a uniformly positive outcome of improved skin integrity, decreased appearance of fine wrinkling, and decreased erythema in the management of occupational dry skin and irritant contact dermatitis. PMID- 12548257 TI - Adverse reactions associated with an alcohol-based hand antiseptic among nurses in a neonatal intensive care unit. AB - BACKGROUND: Alcohol-based hand antiseptics are strongly recommended in the 2002 Centers for Disease Control and Prevention's hand-hygiene guideline. In a study comparing 2 hand-hygiene regimes, an alcohol-based (61% ethyl) antiseptic and a detergent containing 2% chlorhexidine gluconate in 2 neonatal intensive care units, we noted adverse reactions associated with the alcohol-based antiseptic. METHODS: A prospective study was conducted of the skin condition of 58 nurses using an alcohol-based product from March 2001 to January 2002. Adverse reactions to the alcohol-based product were noted and the Fisher exact test was used to determine factors associated with these reactions. Nurses with reactions to the alcohol product who were available to follow-up were patch tested to the product. RESULTS: Of 58 (1.1/100 nursing mo) nurses, 7 were evaluated by occupational health services for dermatologic symptoms that varied from mild to severe after use of the alcohol product, but 4 of 7 have resumed use. Nurses who had adverse reactions develop had been employed on the study unit and in the nursing profession for significantly less time than those with no reactions (P =.037 and P =.002, respectively), and were significantly more likely to report a history of itchy, sore skin (P =.047). A positive patch-test result was noted in 3 of 4 nurses with a previous reaction to the product. CONCLUSION: This case series will alert users in the United States and elsewhere to the nature of reactions to alcohol products and how these reactions differ from reactions to traditional hand antiseptic products. PMID- 12548258 TI - Microbial contamination of enteral feed administration sets in a pediatric institution. AB - BACKGROUND: Enteral feeding tubes have been associated with outbreaks of antimicrobial-resistant organisms, but the pathogenesis of this association has not been investigated. We hypothesized that the enteral feed administration sets become colonized externally by microbes grown from the enteral tube hub, and therefore serve as a reservoir of organisms that can be crosstransmitted. METHODS: We conducted a prospective observational cohort pilot study, obtaining bacterial cultures from the external enteral feed administration set and from the hub of nasogastric, gastric, or gastrojejunal tubes in children receiving enteral feeding while hospitalized in a tertiary care pediatric hospital. RESULTS: Thirty six of 37 hubs cultured had bacterial growth. Twenty-nine of 36 administration sets (78%) sampled had at least 1 microbe isolated that was also cultured from the hub. No significant risk factors for colonization were identified. CONCLUSIONS: Enteral feed administration sets are frequently colonized by organisms in the enteral tube hub. These sets can serve as a reservoir of organisms that can be crosstransmitted between patients. Adherence to Standard Precautions is critical when handling enteral feeding apparatuses. PMID- 12548259 TI - Clostridium difficile infection and concurrent vancomycin-resistant Enterococcus stool colonization in a health care worker: case report and review of the literature. AB - Clostridium Difficile diarrhea was noted in a previously healthy health care worker from the study institution after receiving oral clindamycin therapy; the worker also had vancomycin-resistant Enterococcus stool colonization. Health care workers should be aware that antibiotic therapy may place them at increased risk for colonization and infection with nosocomial pathogens such as Clostridium difficile and vancomycin-resistant Enterococcus. PMID- 12548260 TI - The case of the cold thermometers. AB - BACKGROUND: A temporal artery thermometer was introduced at our hospital. Immediately staff noted subnormal temperature readings. METHODS: Three serial evaluations of patient temperatures were done: August 2000 (initial), September 2000 (posteducation), and December 2000 (after removal of temporal artery thermometer). RESULTS: In the initial review, 89% of the readings were < 98.6 degrees F. Comparison of the readings from the last and first surveys showed a statistical increase in the proportion of readings > or =100.0 degrees F and decrease of readings <98.6 degrees F (P <.001). CONCLUSIONS: Our experience underscores the importance of performance data for quality monitoring of new products. PMID- 12548261 TI - A new potential route of tick-transmitted bacterial diseases to man and animals? PMID- 12548262 TI - Contaminated operating room boots. PMID- 12548263 TI - Enterococcus faecalis arterial line infection. PMID- 12548264 TI - There is a more efficient way... PMID- 12548266 TI - Intervention to decrease emergency department crowding: does it have an effect on return visits and hospital readmissions? AB - STUDY OBJECTIVES: We evaluate the effect of a multifaceted intervention to decrease emergency department crowding on the incidence of return visits to the ED or a hospital ward. The intervention included increased emergency physician coverage, the designation of physician coordinators, and new hospital policies regarding laboratory, consultation, and admission procedures. METHODS: The incidence of return visits within 7 days of discharge was estimated in samples from 2 populations (ie, patients discharged from the ED and patients discharged from the hospital) and during a 12-month period before and a 12-month period after the implementation of the intervention. Return visits were categorized into the following groups: (1) scheduled or not and (2) related or not to initial visit. Logistic regression was used in subsamples to assess the effect of the intervention while controlling for potential confounders. By using information from the provincial medical services database, variation between the periods before and after implementation of the intervention in the incidence of return visits to any ED was compared between the study hospital and 2 external control hospitals. RESULTS: No difference was found in the incidence of return visits between the periods before and after implementation of the intervention, either for patients discharged from the ED (all returns: 11.0% versus 12.4%, 95% confidence interval on difference [CID] -1.5% to 4.3%; unscheduled-related returns: 6.5% versus 5.8%, 95% CID -2.8% to 1.6%) or the hospital (all returns: 6.8% versus 6.6%, 95% CID -2.5% to 2.1%; unscheduled-related returns: 4.2% versus 4.0%, 95% CID -2.0% to 1.7%). This lack of effect remained even after controlling for potential confounders. Variation between the periods before and after implementation of the intervention in the incidence of return to any ED was similar in the 3 hospitals examined. CONCLUSION: Our successful hospital intervention to decrease crowding reduced the mean length of stay for patients discharged from the ED from 13.8 to 5.9 hours, without resulting in increased return visits to the ED or hospital readmission. PMID- 12548267 TI - An inexpensive modification of the laboratory computer display changes emergency physicians' work habits and perceptions. AB - STUDY OBJECTIVE: Little is known about how the availability of laboratory data affects emergency physicians' practice habits and satisfaction. We modified our clinical information system to display laboratory test status with continuous updates, similar to an airport arrival display. The objective of this study was to determine whether the laboratory test status display altered emergency physicians' work habits and increased satisfaction compared with the time period before implementation of laboratory test status. METHODS: A retrospective analysis was performed of emergency physicians' actual use of the clinical information system before and after implementation of the laboratory test status display. Emergency physicians were retrospectively surveyed regarding the effect of laboratory test status display on their practice habits and clinical information system use. Survey responses were matched with actual use of the clinical information system. Data were analyzed by using dependent t tests and Pearson correlation coefficients. The study was conducted at a university hospital. RESULTS: Clinical information system use by 46 emergency physicians was analyzed. Twenty-five surveys were returned (71.4% of available emergency physicians). All emergency physicians perceived fewer clinical information system log ons per day after laboratory test status display. The actual average decrease was 19%. Emergency physicians who reported the greatest decrease in log ons per day tended to have the greatest actual decrease (r =-0.36). There was no significant correlation between actual and perceived total time logged on (r =0.08). In regard to effect on emergency physicians' practice habits, 95% reported increased efficiency, 80% reported improved satisfaction with data access, and 65% reported improved communication with patients. CONCLUSION: An inexpensive computer modification, laboratory test status display, significantly increased subjective efficiency, changed work habits, and improved satisfaction regarding data access and patient communication among emergency physicians. Knowledge of the test queue changed emergency physician behavior and improved satisfaction. PMID- 12548268 TI - Interobserver agreement in emergency department triage. AB - STUDY OBJECTIVES: We measure the interobserver reliability of the triage process, examine the effect of vital signs on the triage process, and provide a context for the prior observation of poor interobserver agreement between in-person and telephonic interviews. METHODS: We performed a prospective observational study using a randomized crossover design at a university teaching hospital emergency department. Patients were eligible if they spoke English, were not presenting for a reevaluation, and were unlikely to be harmed by the delay created by a second triage interview. Every eligible patient underwent 2 independent, sequential, in person ED intake interviews conducted by experienced ED triage nurses. After taking a history, each nurse chose 1 of 5 hypothetic triage designations (ED by 911, ED within 2 hours, see a physician within 8 hours, see a physician within 24 hours, or home care-see a physician in >24 hours) and, after being told the patient's vital signs, again selected a designation. RESULTS: Three hundred sixty three patients presented during the study period: 113 were ineligible, 34 were missed by the investigators, and 15 refused to participate. Nineteen nurses participated in the triage of the 201 study patients. Agreement between 2 in person designations made without knowledge of vital signs was poor (percent agreement 53%; kappa=0.30; tau(b)=0.50). Knowledge of vital signs did not improve agreement (percent agreement 49%; kappa=0.25; tau(b)=0.45). CONCLUSION: There was poor interobserver agreement between certified triage nurses using a 5-item triage scale designed for telephonic triage. These findings suggest that only a small portion of the poor interobserver agreement observed in a prior study of telephonic versus in-person triage can be attributed to the use of the telephone. PMID- 12548269 TI - The value of parental report for diagnosis and management of dehydration in the emergency department. AB - STUDY OBJECTIVES: We define the predictive value of parents' computer-based report for history and physical signs of dehydration for a primary outcome of percentage of dehydration (fluid deficit) and 2 secondary outcomes: clinically important acidosis and hospital admission. We also sought to compare the reports of physical signs related to dehydration made by parents and nurses. METHODS: We performed a prospective observational trial in an urban pediatric emergency department. A convenience sample of parents completed a computer-based interview covering historical details and physical signs (ill appearance, sunken fontanelle, sunken eyes, decreased tears, dry mouth, cool extremities, and weak cry) related to dehydration. Nurses independently completed an assessment of physical signs for enrolled children. The primary outcome was the degree of dehydration (fluid deficit), which was defined as the percentage difference between initial ED weight and stable final weight after the illness. Secondary outcomes included clinically important acidosis (defined as a serum CO(2) value of 75, were randomly selected from the Negev population. RESULTS: Dietary intake of energy, fat, carbohydrates, vitamins E, C and B(1) were significantly lower for people aged 75 and older compared with people aged 65-74. Low energy intake was associated with lower subjective health status for men (P<0.01), poor appetite (P<0.01) and more gastrointestinal problems (P<0.05) for women and lower snack consumption (P<0.01) for both genders. In a multivariate model, low energy intake was associated with low appetite and higher use of medication for both sexes and with frequency of eating alone and not consuming snacks for men. CONCLUSIONS: Dietary intake is lower among individuals older than 75 than in 65-74 y old. Risk factors for low intake include poor appetite and health status, gastrointestinal problems, and eating alone. Snacking enhanced dietary intake and should be encouraged in this group. PMID- 12548293 TI - A digestive tolerance study of maltitol after occasional and regular consumption in healthy humans. AB - AIM: We aimed to evaluate the gastro-intestinal tolerance to an indigestible bulking sweetener containing sugar alcohol using a double-blind random cross-over study. METHOD: In order to simulate their usual pattern of consumption, 12 healthy volunteers ingested maltitol or sucrose throughout the day, either occasionally (once a week for each sugar, first period) or regularly (every day for two 9 day periods, second period). In both patterns of consumption, daily sugar doses were increased until diarrhea and/or a grade 3 (ie severe) digestive symptom occurred, at which the dose level was defined as the threshold dose (TD). RESULTS: In the first period (occasional consumption), the mean TD was 92+/-6 g with maltitol and 106+/-4 g with sucrose (P=0.059). The mean intensity of digestive symptoms was 1.1 and 1.3, respectively (P=NS). Diarrhea appeared in six and one subjects respectively (P=0.035). In the second period (regular consumption), the mean TD was 93+/-9 g with maltitol and 113+/-7 g with sucrose (P=0.008). The mean intensity of digestive symptoms was 1.7 and 1.2, respectively (P=NS). However, diarrhea appeared in eight and three subjects, respectively (P=0.04). Maltitol and sucrose TDs between the two periods were not different. CONCLUSIONS: Under our experimental conditions, in comparison to sucrose: (a) occasional or regular consumption of maltitol is not associated with severe digestive symptoms; (b) in both patterns of maltitol consumption, diarrhea frequency is higher, but it appeared only for very high doses of maltitol, much greater than those currently used; (c) maltitol does not lead to intestinal flora adaptation after a 9 day period of consumption. PMID- 12548295 TI - Consumption of black currants, lingonberries and bilberries increases serum quercetin concentrations. AB - OBJECTIVE: To study serum quercetin concentrations of subjects consuming berries or habitual Finnish diets. DESIGN: Randomized parallel dietary intervention. SUBJECTS: Forty healthy men (age 60 y). INTERVENTION: Twenty subjects consumed 100 g/day of berries (black currants, lingonberries and bilberries) for 8 weeks. Twenty subjects consuming their habitual diets served as controls. Fasting blood samples were obtained 2 weeks prior to the study, at baseline, and at 2, 4 and 8 weeks. Intake of quercetin was assessed from 3 day food records collected at baseline and at 8 weeks. RESULTS: The serum quercetin concentrations were significantly higher in the subjects consuming berries compared to the control group (P=0.039 ANCOVA with repeated measures). During the berry consumption period the mean serum concentrations of quercetin ranged between 21.4 and 25.3 micro g/l in the berry group, which was 32-51% higher compared with the control group. According to 3 day food records, there was no difference in quercetin intake at baseline, but at 8 weeks the intake was 12.3+/-1.4 mg/day (mean+/ s.e.m.) in the berry group and 5.8+/-0.6 mg/day in the control group (P=0.001). CONCLUSIONS: The results indicate that the berries used in this study are a good source of bioavailable quercetin. PMID- 12548294 TI - Oral glutamine in paediatric oncology patients: a dose finding study. AB - OBJECTIVE: The purpose of this study was to determine the most appropriate dose of oral glutamine to use in a further clinical study in paediatric oncology patients. DESIGN: This was a phase I, pharmokinetic study. SETTING: The study was carried out at The Yorkshire Regional Centre for Paediatric Oncology and Haematology, St James's University Hospital, Leeds, UK. SUBJECTS: Thirteen patients undergoing treatment for paediatric malignancy participated in this study. All 13 completed the study. INTERVENTIONS: The most appropriate dose was determined by patient acceptability and by plasma glutamine and ammonia levels measured at timed intervals after ingestion of a single glutamine dose. RESULTS: Doses of 0.35, 0.5 and 0.65 g/kg were well tolerated with no untoward plasma glutamine and ammonia levels. One patient was recruited to a higher dose of 0.75 g/kg, but the plasma glutamine and ammonia levels peaked at 2601 and 155 micro mol/l, respectively. The ammonia level was greater than the acceptable upper limit. It was difficult to disperse the glutamine adequately at this dose, resulting in the suspension being found to be unpalatable and therefore no further patients were recruited at this dose. CONCLUSION: It was concluded that 0.65 g/kg is a safe dose of glutamine to use in a clinical study in paediatric oncology patients. PMID- 12548296 TI - A modern day holy anorexia? Religious language in advertising and anorexia nervosa in the West. AB - OBJECTIVE: In this review we comment on the media, in particular advertisements and the internet, using religious and moralistic motifs in connection with eating and their possible role in promoting modern-day 'nutritional' consumerism and fundamentalism (diet, discipline and discipleship). METHOD: We have reviewed historically the concept of holy anorexia and have analyzed the pictorial and language content of some representative advertisements. RESULTS: Words used include 'temptation', 'decadent', 'heaven' and 'purity' and suggest a connection with eating and moral values. They also imply eating as either heaven or sin, to be yielded to in both circumstances. CONCLUSIONS: Such messages identifying eating with guilt or reward could precipitate 'atypical' eating disorders in vulnerable subjects, invoking the historical associations with extreme religiosity and anorexia. It is not clear how these 'fundamentalist' trends developed but health educators should be aware of them in dealing with at-risk subjects for eating disorders. PMID- 12548297 TI - The burden of anemia among women in India. AB - OBJECTIVE: This research investigates the prevalence and determinants of anemia among women in Andhra Pradesh. We examined differences in anemia related to social class, urban/rural location and nutrition status body mass index (BMI). We hypothesized that rural women would have higher prevalence of anemia compared to urban women, particularly among the lower income groups, and that women with low body mass index (BMI; <18.5 kg/m(2)) would have a higher risk compared to normal or overweight women. DESIGN: The National Family Health Survey 1998/99 (NFHS-2) provides nationally representative cross-sectional survey data on women's hemoglobin status, body weight, diet, social, demographic and other household and individual level factors. Ordered logit regression analyses were applied to identify socio-economic, regional and demographic determinants of anemia. SETTING: Andhra Pradesh, a southern Indian state. SUBJECTS: A total of 4032 ever married women aged 15-49 from 3872 households. RESULTS: Prevalence of anemia was high among all women. In all 32.4% of women had mild (100-109.99 g/l for pregnant women, 100-119.99 for non-pregnant women), 14.19% had moderate (70-99.99 g/l), and 2.2% had severe anemia (<70 g/l). Protective factors include Muslim religion, reported consumption of alcohol or pulses, and high socioeconomic status, particularly in urban areas. Poor urban women had the highest rates and odds of being anemic. Fifty-two percent of thin, 50% of normal BMI, and 41% of overweight women were anemic. CONCLUSIONS: New program strategies are needed, particularly those that improve the overall nutrition status of women of reproductive ages. This will require tailored programs across socio-economic groups and within both rural and urban areas, but particularly among the urban and rural poor. PMID- 12548298 TI - Acute effect of high-calcium milk with or without additional magnesium, or calcium phosphate on parathyroid hormone and biochemical markers of bone resorption. AB - OBJECTIVE: To assess whether there are any differences in the postprandial physiological responses to apple drink (control), calcium phosphate (tricalcium phosphate, TCP) and high-calcium skim milk (HCSM) with or without additional magnesium in postmenopausal women. DESIGN: Randomized, controlled, cross-over. Measurements after overnight fast before each drink, and subsequently every hour for 8 h postprandially. SETTING: Human Nutrition Studies Laboratory, Milk and Health Research Centre, Massey University, Palmerston North, New Zealand. SUBJECTS: Twenty-one healthy postmenopausal women. INTERVENTION: Four drinks, each 400 ml. (1) Apple drink (25% fruit juice). (2) TCP dispersed in water containing 1200 mg Ca. (3) HCSM containing 1200 mg Ca and 65.5 mg Mg. (4) HCSM containing 1200 mg Ca and 172 mg Mg. RESULTS: There was no difference in baseline serum calcium, PTH or C-telopeptide levels between drinks. There were no overall differences in serum calcium after apple or after either milk, but after TCP serum calcium increased from a baseline value of 2.12+/-0.08 to a mean peak of 2.21+/-0.12 mmol/l (s.d.) (P=0.0001) after 2 h. There were no significant differences in serum PTH after either apple or HCSM+Mg. In contrast, after TCP, serum PTH decreased from 2.76+/-0.69 to a mean nadir of 2.23+/-0.65 pmol/l (P=0.0001) after 1 h, and after HCSM, it decreased from 2.71+/-0.78 to a mean nadir of 2.51+/-0.87 pmol/l (P=0.007) after 2 h. Serum C-telopeptides decreased after each drink, reaching nadirs after 5 h. At this time the serum values for each of the high calcium drinks were not different from each other, but were significantly less than for apple (P=0.001 for each), being 0.22+/-0.09 ng/ml for apple, 0.15+/-0.08 for TCP, 0.14+/-0.07 for HCSM and 0.16+/-0.07 for HCSM+Mg. CONCLUSION: Despite differences in serum calcium and PTH responses to the three high-calcium drinks that we tested, there was no distinguishable difference in serum C-telopeptides between high calcium drinks. PMID- 12548299 TI - Prevalence of risk of undernutrition is associated with poor health status in older people in the UK. AB - OBJECTIVES: To establish the prevalence of the risk of undernutrition, using criteria similar to those used by the Malnutrition Advisory Group (MAG), in people aged 65 y and over, and to identify relationships between risk of undernutrition and health and demographic characteristics. DESIGN: A cross sectional nationally representative sample of free-living and institutionalized older people in the UK (65 y of age and over). Secondary analysis of the National Diet and Nutrition Survey based on 1368 people aged 65 y and over. RESULTS: About 14% (21% in those living in institutions) were at medium or high risk of undernutrition based on a composite measure of low body mass index and recent reported weight loss. Having a long-standing illness was associated with a statistically significantly increased risk of undernutrition (odds ratio: men 2.34, 95% CI 1.20-4.58; women 2.98; 1.58-5.62). The risk of undernutrition increased: in women reporting bad or very bad health status; in men living in northern England and Scotland; for those aged 85 y and older; for those hospitalized in the last year, and those living in an institution. Lower consumption of energy, meat products or fruit and vegetables and lower blood measures of zinc, vitamins A, D, E and C were associated with statistically significantly increased risk of undernutrition. CONCLUSIONS: A substantial proportion of the older population of the UK is at risk of undernutrition. High risk subjects are more likely to have poorer health status. It is unlikely that the individuals at high risk are being detected currently, and therefore effective care is not being provided, either in the community or in institutions. PMID- 12548300 TI - Serum transferrin receptor in children with respiratory infections. AB - OBJECTIVE: To study the effect of infection on iron status in children suffering from acute, mild or severe respiratory infections and to determine the nature of anemia in infection using serum transferrin receptor (sTfR) levels. DESIGN: Forty three children aged between 3 and 5 y with no evidence of infection and receiving iron supplements in the preceding 100 days served as controls. Twenty-one children with mild upper respiratory infection and 94 children hospitalized for acute pneumonia constituted the experimental group. Hemoglobin (Hb), sTfR and serum ferritin were estimated in all the children at the time of diagnosis and again on the 15th and 30th days after the infection in those who were available for follow-up. RESULTS: Mean (95% CI) sTfR was 6.08 (5.1-7.1) mg/l in healthy non anemic children. Upper respiratory infection had no impact on Hb or sTfR but it significantly elevated serum ferritin levels. Eighty-three percent of the children with pneumonia had Hb less than 110 g/l at the time of diagnosis and had elevated mean sTfR, 18.0 (15.7-20.3) mg/l. There was a decline in mean sTfR by the 15th day of infection to 14.3 (11.3-17.4) mg/l with further rise to 22.9 (13.0-31.9) mg/l by 30 days. Serum ferritin was significantly elevated at the time of diagnosis (85.9; 71.1-100.8 micro g/l) as well as at 15 days (89.1; 68 110.1 micro g/l) with a decline by 30 days. CONCLUSIONS: Severe lower respiratory infection exaggerates iron-deficient erythropoiesis by blocking release of iron from the storage pools. sTfR may not be a sensitive and specific tool of assessing true iron status of children exposed to severe infections. PMID- 12548301 TI - Folate intake, plasma folate and homocysteine status in a random Finnish population. AB - OBJECTIVE: To assess the folate status of Finnish adults using plasma folate and homocysteine as biomarkers and to evaluate dietary and supplementary folate intakes. MATERIALS AND METHODS: Plasma folate, vitamin B(12) and total homocysteine (tHcy) were determined in a random sample of 643 subjects aged 25-74 y living in the Helsinki area. The methylenetetrahydrofolate reductase (MTHFR) genotypes were analyzed from a subsample (n=394). Dietary intake data by 24 h recall and use of vitamin supplements were collected. RESULTS: Plasma folate was normal (>/=5 nmol/l) in 99% of subjects and optimal (>/=8 nmol/l) in terms of a minimum tHcy in 90%. Mean plasma folate of non-supplement users was 13.7 and 12.9 nmol/l and tHcy 11.3 and 9.2 micro mol/l for men and women, respectively. Elevated tHcy (>14 micro mol/l) was found in 11% of subjects. Homozygote frequency for MTHFR genotype TT was 5.0% and their plasma tHcy was 14.8 micro mol/l compared to the mean of the other subjects, 10.5 micro mol/l, P<0.05. The mean dietary folate intake was 241 micro g/day (29 micro g/MJ of energy) for men and 205 micro g/day (33 micro g/MJ) for women, respectively. The main dietary sources of folate were vegetables 12%, wholemeal ryebread 11%, fruits 10%, and potato 10%. Regular supplement users (n=97) received on average 207 micro g folic acid per day from supplements. CONCLUSIONS: The folate status of Finnish adults seems to be adequate according to energy adjusted folate intake, plasma folate and homocysteine. The MTHFR homozygote frequency was low compared to other countries. Regular use of supplementary folic acid less than 300 micro g increased plasma folate, but supplemental folic acid over 300 micro g was required to lower tHcy values significantly. PMID- 12548302 TI - Long-chain polyunsaturated fatty acids at birth and cognitive function at 7 y of age. AB - OBJECTIVE: During the central nervous system (CNS) growth spurt, rapid accretion of long chain polyunsaturated fatty acids (LCPUFA) takes place. This particularly concerns docosahexaenoic acid (DHA, 22:6n-3) and arachidonic acid (AA, 20:4n-6), which are thought to play important roles in CNS development and function. The aim of this study was to investigate the relationship between cognitive performance at 7 y of age and LCPUFA levels in umbilical venous plasma phospholipids, representing the prenatal fatty acid availability, and in plasma phospholipids sampled at 7 y. DESIGN: As part of a follow-up study, the cognitive performance of 306 children, born at term, was assessed at 7 y of age with the Kaufman Assessment Battery for Children. Backward stepwise regression analysis was used to study the relationship between the outcomes and LCPUFA status. Social class, maternal intelligence and parenting skills were included as covariables, among others. RESULTS: Results show no significant association with either DHA or AA at birth and the cognitive performance at 7 y of age. The LCPUFA levels at 7 y were not associated with these outcomes either. Consistent with the literature, significant relationships were found between cognitive outcome measures and maternal education, maternal intelligence and the child's birthweight. CONCLUSIONS: In conclusion, our results do not provide evidence for a positive association between cognitive performance at 7 y and LCPUFA status at birth or at 7 y of age. PMID- 12548303 TI - Effect of c-reactive protein and interleukins blood levels in postsurgery arginine-enhanced enteral nutrition in head and neck cancer patients. AB - OBJECTIVE: It is known that the immune system is frequently affected in patients with head and neck cancer. Although immune dysfunction could be multifactorial, this immune system may be modulated by specific nutritional substrates, such as arginine. The aim of our study was to evaluate the effect of enteral nutrition supplemented with arginine on c-reactive protein (CRP), interleukin 6 (IL-6) and tumour necrosis factor (TNFalpha) in surgical head and neck cancer patients. DESIGN: Randomized trial. SETTING: Tertiary care. SUBJECTS: A population of 36 patients with oral and laryngeal cancer were enrolled. INTERVENTIONS: At surgery patients were randomly allocated to two groups: (a) patients receiving an enteral diet supplements with arginine and dietary fibre (group I, n=18); (b) patients receiving an isocaloric, isonitrogenous enteral formula (group II, n=18). Perioperatively and on postoperative day 5 the following parameters were evaluated: serum values of prealbumin, transferrin, albumin, total number of lymphocytes, interleukin 6, tumour necrosis factor alpha and c-reactive protein. RESULTS: The mean age was 59.6+/-10.9 y (two females/34 males). No significant intergroup differences in the trend of the three plasma proteins and weight were detected. CRP decreased in both groups (group I: 152.9+/-76.9 vs 68.9+/-82.5 mg/dl; P<0.05; and group II: 105.9+/-92 vs 43.6+/-59.1 mg/dl; P<0.05). Interleukin 6 did not change (group I: 16.3+/-12.3 vs 35.6+/-83.4 pg/ml; NS; and group II: 22.8+/-40 vs 9.9+/-17.7 pg/ml; NS). TNFalpha did not show any differences (group I: 4.6+/-1.6 vs 5.1+/-1.5 pg/ml; NS; and group II: 8.8+/-6.1 vs 5.8+/-1.7 pg/ml; NS). Lymphocytes increased in both groups (group I: 1405.6+/ 517 vs 1634+/-529 x 10(6)/ml; P<0.05; and group II: 1355+/-696 vs 1561+/-541 x 10(6)/ml; P<0.05). CONCLUSIONS: Enhanced formula did not change IL6 and TNFalpha levels. Further studies are needed to determine whether route of nutrition or type of formula is the key in these patients. PMID- 12548304 TI - Dietary supplements of soya flour lower serum testosterone concentrations and improve markers of oxidative stress in men. AB - OBJECTIVE: We examined the effects on serum sex steroids, lipids and markers of oxidative stress of supplementing the diets of healthy male volunteers with scones made with soya flour. DESIGN: A randomized placebo controlled cross-over trial. SETTING: University Hospital of Wales. SUBJECTS: Twenty volunteers recruited by advertisement. INTERVENTIONS: Male volunteers ate three scones a day in addition to their normal diet for a period of 6 weeks. The scones were made with either wheat or soya flour (containing 120 mg/day of isoflavones). Blood was analysed for sex steroids (testosterone, dihydro-testosterone, oestradiol, oestrone, sex hormone binding globulin, albumin and the concentration of non protein bound sex steroids were calculated), lipid profile (total cholesterol, high density lipoprotein cholesterol and triglycerides) and measures of oxidative stress (hydroperoxides, susceptibility of LDL to oxidation with copper and myeloperoxidase). RESULTS: The volunteers' mean age was 35.6 (s.d. 11.2) y. Total serum testosterone fell in volunteers taking the soya scones (19.3-18.2 nmol/l; 95% CI 1.01, 1.12; P=0.03). No significant changes were seen in the concentrations of the other serum sex steroids, albumin or sex hormone binding globulin throughout the study. Significant improvements in two of the three markers of oxidative stress were seen in volunteers taking soya scones. Lag time for myeloperoxidase rose from 55.0 to 68.0 min (95% CI -16.0, -3.5; P=0.009) and the presence of hydroperoxides decreased from 2.69 to 2.34 micro mol/l (95% CI 0.12, 0.71; P=0.009). There were no changes seen in serum triglycerides or cholesterol. CONCLUSIONS: We have shown that soya supplements reduce serum testosterone and improve markers of oxidative stress. These findings provide a putative mechanism by which soya supplements could protect against prostatic disease and atherosclerosis. Further dietary studies with clinical end points are warranted. PMID- 12548305 TI - Comparison of telephone vs face-to-face interviews in the assessment of dietary intake by the 24 h recall EPIC SOFT program--the Norwegian calibration study. AB - OBJECTIVE: To compare food group intakes in grams, total energy and energy contribution from macronutrients between two random samples of women, using a standardized computer program (EPIC SOFT) with either a face-to-face or a telephone 24 h recall interview design. METHODS: Two groups of Norwegian women living in Tromso were drawn at random from the Norwegian Women and Cancer Study (NOWAC) cohort. The EPIC SOFT computer program was used to conduct single 24 h dietary recalls either by telephone or face to face. For the latter, 160 women were invited, of whom 111 responded positively (crude response rate=69.4%) and 102 were interviewed. For the telephone option, 180 were invited to participate, 109 responded positively (crude response rate=60.6%) and 103 were interviewed. RESULTS: The two groups of women were similar with respect to age, body mass index, basal metabolic rate, smoking, education, physical activity and special diet status. No statistically significant difference in dietary intake was found between interviews conducted by telephone and face-to-face, except for 'egg and egg products' (P<0.01), for which intake was higher in the telephone group. Total energy intake differed, but the difference was attributable to one interviewer. The distribution of energy-contributing nutrients was not significantly different between groups, except for reported energy percentage from protein (P=0.02), which was significantly higher in the face-to-face group. CONCLUSION: The telephone vs face-to-face interview design did not influence recalled diet when using the EPIC SOFT program. PMID- 12548306 TI - Associations between reproduction and maternal body weight: examining the component parts of a full reproductive cycle. AB - OBJECTIVE: Many transitional societies currently face both extremes of nutritional status, undernutrition and overnutrition. Women of reproductive age are at high risk of these conditions. The purpose of this review is to consider evidence for relationships between reproduction and nutritional status in women from societies of varying economic development, using body weight or weight-for height as indicators of maternal nutritional status. DESIGN: The conceptual framework guiding this review is that the duration of the reproductive cycle varies as a function of its component parts, which include (i) pregnancy, (ii) lactation, (iii) the non-pregnant/non-lactating (NP/NL) interval or, possibly, (iv) an overlap between lactation and next pregnancy. All component parts of the complete cycle vary in length and are associated with changes in nutritional status. A variety of factors ('proximal and distal determinants') influence the duration of the component parts of the reproductive cycle. This framework is used to examine current knowledge of changes in maternal nutritional status during each of these parts. RESULTS: Women in affluent societies retain some weight with each pregnancy, beyond that of non-pregnant women. Women in less affluent societies retain less weight with each pregnancy. During lactation, women in both affluent and less affluent societies experience only modest weight loss. During the NP/NL interval, women in affluent societies tend to gain weight, whereas weight of women in less affluent societies is likely to fluctuate. CONCLUSIONS: We conclude that there is a dearth of information on certain parts of the cycle, particularly the periods of overlap of lactation with pregnancy and the NP/NL interval. PMID- 12548307 TI - Socio-economic status, dietary intake and 10 y trends: the Dutch National Food Consumption Survey. AB - OBJECTIVE: To study differences in dietary intake between adults with different socioeconomic status (SES) and trends over time. DESIGN: Cross-sectional study based on data of three Dutch National Food Consumption Surveys (DNFCS-1 1987/88; DNFCS-2 1992; DNFCS-3 1997/98), obtained from a panel by a stratified probability sample of the non-institutionalized Dutch population. SUBJECTS: A total of 6008 men and 6957 women aged 19 y and over. METHODS: Dietary intake was assessed with a 2 day dietary record. Background information was obtained by structured questionnaire. Sociodemographic variables were available from panel information. SES, based on educational level, occupation and occupational position was categorized into (very) low, middle and high. Analysis of variance with age as covariable was used to explore the effects of SES on dietary intake and anthropometry. Statistical tests for trend were carried out with models in which week-weekend-day effects and an interaction term of time with SES were also included. RESULTS: The prevalence of obesity and skipping of breakfast was higher among people with a low SES. In all three surveys, subjects in the (very) low SES group reported having a higher consumption of potatoes, meat and meat products, visible fats, coffee and soft drinks (men only). Subjects with a high SES reported consuming more vegetables, cheese and alcohol. As regards nutrients, in all surveys a higher SES was associated with higher intake of vegetable protein, dietary fibre and most micronutrients. A higher SES was also associated with a lower fat intake but the differences between social classes were rather small and not consistent when the contribution of alcohol to energy intake was taken into account. CONCLUSION: In general, dietary intake among subjects in higher SES groups tended to be closer to the recommendations of the Netherlands Food and Nutrition Council and this phenomenon was quite stable over a period of 10 y. PMID- 12548308 TI - Para-aminobenzoic acid (PABA) used as a marker for completeness of 24 hour urine: effects of age and dosage scheduling. AB - OBJECTIVE: To examine the age dependency of the urinary para-aminobenzoic acid (PABA) excretion, and if a delayed PABA excretion can be overcome by advancing intake schedule; and to examine the recovery of PABA in fractionated urinary samples collected during 24 h after single and repeated doses of PABA. DESIGN: Cross-over study with subjects randomized to start with recommended schedule of PABA administration (80 mg at 08:00, 12:00 and 18:00; PABA18) and then an advanced schedule (80 mg at 08:00, 12:00 and 15:00; PABA15) or vice versa. One subgroup of eight subjects collected individual urine specimens for 24 h after a morning dose of 80 mg of PABA, and another subgroup of 10 subjects collected individual urine specimens for 24 h after ingestion of 80 mg of PABA three times at mealtimes. SUBJECTS: Employees and relatives from the Danish Food Administration. SETTING: Ninety-nine healthy volunteers (61 females and 38 males) aged 30-91 y. RESULTS: Linear regressions for PABA15 and PABA18 demonstrate significantly less recovery with age (PABA15: r(2)=0.1784, P=0.0002; PABA18: r(2)=0.1273, P=0.0019). Linear regression of DeltaPABA (PABA15-PABA18) with age showed the best fit line to be horizontal (slope -0.0066, P=0.89; 95% CI -0.1046, 0.0915) and with a Y-intercept not significantly different from 0 (1.575; 95% CI 4.176, 7.326). In this population the lower limit for complete 24 h urine collection was 79.2%. After a single dosage of 80 mg PABA 70-85% was recovered after 8 h. Within 16 h after ingestion of 240 mg PABA at recommended hours the lowest acceptable recovery (78.1%) was reached. CONCLUSION: There is a gradual decline of PABA recovery with age that cannot be overcome by advancing the dosage schedule. Because of a lower delimiting PABA recovery for the elderly, some 24 h collections in this age group will be rejected unjustly (false-negatives). Also, with the currently recommended dosage schedule (PABA taken with the main meals) the risk of false-positive 24 h urine collections prevails. With refinement of the PABA test procedure, ie employing a specific analytical method and age dependent cut-off values, the test may achieve a higher specificity and sensitivity. PMID- 12548309 TI - Social inequalities and children's height in Trinidad and Tobago. AB - OBJECTIVE: The aim of the study was to report the association of socio-economic factors with child's height. DESIGN: Cross-sectional study based on a representative national sample of government schools. SETTING: Trinidad and Tobago in 1999. SUBJECTS: A total of 2608 boys and girls mean age 5.8 y, range 4.38-6.99 y and 3080 mean age 8.6 y, range 7.00-10.44 y olds. OUTCOME: Measurement of height and a questionnaire completed by parents. In the analysis height was expressed as standard deviation scores (s.d.s.) based on the British height curves (1990) or height below -1.5 s.d.s. RESULTS: Ethnicity, parental heights, birthweight, maternal age at child's birth and number of children in the family were the main factors associated with children's height. Lack of piped water supply in the home was the only socio-economic factor consistently associated with height (mean difference in s.d.s. adjusted only for age group, gender and ethnicity -0.192, 95% CI -0.257 to -0.127 and in addition adjusted for the variables listed above -0.080, 95% CI -0.141 to -0.019). Parental education, household overcrowding and employment status were weakly associated with height in the partially adjusted model only. Analysis of severe growth failure gave similar results. CONCLUSION: The impact of socio-economic factors on height is marginal in Trinidad and Tobago. As socio-economic factors may have an impact on a broad range of health indicators, height and rates of undernutrition should not be used as sole criteria for assessing progress in decreasing health differentials caused by social inequalities. PMID- 12548310 TI - Energy stress during pregnancy and lactation: consequences for maternal nutrition in rural Bangladesh. AB - OBJECTIVE: To assess the relationship of energy stress during pregnancy and lactation to maternal body stores in marginally nourished rural Bangladeshi women. SUBJECTS AND METHODS: Two-hundred and fifty-two women were followed from 5 7 months of pregnancy until 6 months postpartum. Energy intake was estimated during pregnancy and at 1, 3 and 6 month(s) postpartum using 24 h dietary recall. Body weight was measured on enrollment, another once or twice during pregnancy, and at 1, 3 and 6 month(s) postpartum. The weekly rates of pregnancy weight gain and postpartum weight changes were determined. Weight and length of the infants were measured at birth and at approximately 1, 3 and 6 month(s). RESULTS: Maternal energy intake at 5-7 months of gestation was 1464+/-416 kcal/day (mean+/ s.d.). Women gained a mean of 200 g/week or a total of 4 kg during the second half of pregnancy. An analysis of maternal weight showed no indication of accrual of fat stores during pregnancy. Dietary energy during lactation exceeded the intake during pregnancy by 248-354 kcal/day. Mothers lost an estimated average of 1 kg of weight during the first 6 months of lactation. The mean (+/-s.d.) birth weight was 2.55+/-0.38 kg, and the prevalence of low birth weight (<2500 g) was 48%. Infants exhibited some catch-up growth only during the first 3 months but overall growth during the first 6 months did not change from their relative status at birth when compared with NCHS reference. CONCLUSIONS: These rural Bangladeshi women failed to gain sufficient weight during the last half of pregnancy to maintain body weight during lactation when the energy demand is high. Poor growth of their primarily breastfed infants raises concern about the adequacy of lactation in this community. PMID- 12548311 TI - Body image of adolescents in a multi-ethnic Caribbean population. AB - OBJECTIVE: To examine the perceptions of body size among adolescents in Trinidad and to determine whether there were ethnic differences. DESIGN: Cross-sectional survey. SETTING: Secondary schools in Trinidad. SUBJECTS: A stratified random sample of 1139 adolescents, aged 14-17 y. MEASUREMENTS: Silhouettes of different body sizes were used to determine perceptions of body size, while weights and heights were measured to determine actual body size. RESULTS: A total of 1090 students (96% response rate) participated, comprising 578 (53%) females. The main ethnic groups were South Asian (49%), African (25%) and persons of mixed ethnicity (23%). The calculated body mass index (BMI) indicated that 14% were thin, 73% normal and 13% overweight; however there was a preponderance of thinness among South Asian males (28%). Whereas 68% of students correctly identified their body size, South Asian males were also more likely to overestimate their body size than the other adolescents, ie they were thinner than they thought. Overall, 64% of the sample was satisfied with their current size, but thin South Asians were more likely to be satisfied with their size than other thin adolescents (P=0.04), while overweight Africans were more likely to be satisfied than other overweight persons (P=0.03). The majority of the sample associated the normal body size with good health, but the majority also associated the overweight and obese silhouettes with wealth. In addition, 40% of them associated the male overweight and obese silhouettes with happiness. CONCLUSION: Although the prevalence of overweight/obesity was not high, the perception of the Trinidadian adolescent that obesity was associated with wealth, and to a lesser extent happiness, and the satisfaction of the overweight African females with their size, were all causes for concern. PMID- 12548312 TI - Adult coeliac patients do tolerate large amounts of oats. AB - OBJECTIVE: The aim of the present study was to investigate whether adult patients with coeliac disease in remission could include large amounts of oats in their daily gluten-free diet for an extended period of time without adverse effects. DESIGN, SUBJECTS AND METHODS: Twenty adult coeliac patients in remission included large amounts of uncontaminated rolled oats in their daily diet for a prolonged period. The examinations, performed four times during the study period, included small bowel endoscopy with biopsies, blood samples (nutritional status, serological analysis), height and body weight, gastrointestinal symptoms and dietary records. Gastrointestinal symptoms and diet were also investigated through unannounced telephone interviews once a month during the study period. RESULTS: No adverse effects of a large intake of oats were seen in small bowel histology, serology nor in nutritional status in the 15 subjects who completed the whole study period. Two of the subjects dropped out because of gastrointestinal symptoms and three for non-medical reasons. The median intake of oats was 93 g/day and the compliance to the oat diet was found to be good. Examinations of the patients after drop-out did not show any deterioration in small bowel histology or nutritional status nor raised levels of antibodies. CONCLUSION: Results from this study indicate that adult patients with coeliac disease in remission can include large amounts of controlled wheat-free rolled oats for an extended period of time without adverse effects. PMID- 12548313 TI - Plant sterol-enriched spread enhances the cholesterol-lowering potential of a fat reduced diet. AB - OBJECTIVE: To determine the effect of a plant sterol-enriched spread on plasma cholesterol concentrations when replacing butter or a standard polyunsaturated spread in a diet containing 30% of energy fat. DESIGN: Parallel butter phase followed by double-blind, randomized, cross-over polyunsaturated spread phases. SETTING: General community. SUBJECTS: Volunteer sample of 50 free-living men and women with mean age (s.d.) 46.7 y (10.5), moderately elevated plasma total cholesterol 5.95 mmol/l (0.78), and body mass index 26.0 (3.9) kg/m(2). INTERVENTION: Participants ate a moderately low-fat diet (30% of energy) for the 11-week intervention. During the first 3 weeks the diet included 20 g per day of butter. Participants were then randomized to replace the butter with 25 g of polyunsaturated spread with or without 2 g of plant sterols for 4 weeks, crossing over in the last 4 weeks to the alternate spread. MAIN OUTCOME MEASURES: Plasma cholesterol and fatty acids. RESULTS: Replacing butter with a standard polyunsaturated fat spread reduced mean plasma total cholesterol concentrations by 4.6% (from 6.09 (0.82) to 5.81 (0.77) mmol/l, P<0.01) and low-density lipoprotein cholesterol by 5.5% (from 3.98 (0.76) to 3.76 (0.74) mmol/l, P<0.05). Replacing butter with a polyunsaturated spread containing plant sterols reduced plasma total cholesterol by 8.9% (from 6.09 (0.82) to 5.55 (0.76) mmol/l, P<0.01) and low density lipoprotein cholesterol by 12.3% (from 3.98 (0.76) to 3.49 (0.72) mmol/l, P<0.01). Plasma high density lipoprotein cholesterol concentration was the same on the three diets. CONCLUSION: In people with moderately raised plasma cholesterol concentrations consuming reduced-fat diets the reduction in plasma total and low-density lipoprotein cholesterol concentrations achieved by replacing butter with a polyunsaturated spread is enhanced by addition of plant sterols. PMID- 12548314 TI - Impact of 'Mad Cow Disease' publicity on trends in meat and total vitamin A consumption in Geneva between 1993 and 2000. AB - BACKGROUND: In March 1996, revelations about the possible risk for humans of the 'mad cow disease' epidemic had a sudden impact on the diets of European populations. OBJECTIVE: To assess changes in meat and nutrient intakes in adults living in Geneva, Switzerland from 1993 to 2000. DESIGN: Independent annual cross sectional surveys (4047 women and 4092 men total). MAIN OUTCOME MEASURE: Dietary habits assessed and compared to baseline (January 1993-April 1996) via validated semi-quantitative food-frequency questionnaire. RESULTS: Women beef abstainers increased from 8.9 to 14.9% in late 1996 (P<0.001) and 13.3% in 1997 (P<0.05); among meat consumers, in late 1996 meat/beef intakes declined -10/-12% (both P<0.05). From 1997 to 2000 most intake levels drifted back toward those at baseline, but chicken intakes were significantly (all P<0.05) greater each year (+19% in 2000 (P<0.001). Consistent but less dramatic changes were observed among men. From late 1996 until 2000, liver abstention was significantly (all P<0.05) greater (women from 60 to 78%; men from 61 to 73% in 2000; (both P<0.001). The only nutrient intakes that decreased significantly (all P<0.05) each year from 1997 through 2000 were retinol and total vitamin A women: -22% (P<0.001); -11% (P<0.05) respectively; men: -16% (P<0.001); -10% (P<0.05) respectively, in 2000). Total vitamin A intakes exceeded the dietary reference intake (DRI) for liver eaters (women 185%, men 153%), but were below the DRI for liver abstainers (women 83%; men 66%) in 2000. CONCLUSION: The decreases in beef and liver consumption since late 1996 led to the discovery of a long-term disparity in the retinol and total vitamin A intakes of liver eaters vs abstainers. PMID- 12548315 TI - Tyrosol and hydroxytyrosol are absorbed from moderate and sustained doses of virgin olive oil in humans. AB - OBJECTIVE: To investigate the absorption of tyrosol and hydroxytyrosol from moderate and sustained doses of virgin olive oil consumption. The study also aimed to investigate whether these phenolic compounds could be used as biomarkers of virgin olive oil intake. DESIGN AND INTERVENTIONS: Ingestion of a single dose of virgin olive oil (50 ml). Thereafter, for a week, participants followed their usual diet which included 25 ml/day of the same virgin olive oil as the source of raw fat. SETTING: Unitat de Recerca en Farmacologia. Institut Municipal d'Investigacio Medica (IMIM). SUBJECTS: Seven healthy volunteers. RESULTS: An increase in 24 h urine of tyrosol and hydroxytyrosol, after both a single-dose ingestion (50 ml) and short-term consumption (one week, 25 ml/day) of virgin olive oil (P<0.05) was observed. Urinary recoveries for tyrosol were similar after a single dose and after sustained doses of virgin olive oil. Mean recovery values for hydroxytyrosol after sustained doses were 1.5-fold those obtained after a single 50 ml dose. CONCLUSIONS: Tyrosol and hydroxytyrosol are absorbed from realistic doses of virgin olive oil. With regard to the dose-effect relationship, 24 h urinary tyrosol seems to be a better biomarker of sustained and moderate doses of virgin olive oil consumption than hydroxytyrosol. PMID- 12548316 TI - Olive-oil consumption and cancer risk. PMID- 12548317 TI - Rebuilding compassionate Canadian health care policy. PMID- 12548318 TI - The management of nausea and vomiting of pregnancy. PMID- 12548320 TI - Impact of the international term breech trial on clinical practice and concerns: a survey of centre collaborators. AB - OBJECTIVE: The purpose of this study was to determine what impact the International Term Breech Trial had had in different settings and to elicit any concerns among collaborators regarding the implementation of a policy of planned Caesarean section for term breech babies. METHODS: We mailed a questionnaire to all Term Breech Trial collaborators. The questionnaire asked 3 open-ended questions about the impact of the trial, about concerns with implementing planned Caesarean section for term breech babies, and about whether information as to the relative costs of planned Caesarean section versus planned vaginal birth would be helpful. Frequencies of responses were calculated for centres in countries classified as having a low or a high national perinatal mortality rate (< or = 20/1000 vs. > 20/1000, respectively) according to the figures published by the World Health Organization in 1996. RESULTS: We received responses from 80 centres in 23 countries. Most centres (92.5%) stated that clinical practice had changed to planned Caesarean section for most or all term breech babies. The majority of centres (66.3%) had no difficulties or concerns with implementing a policy of planned Caesarean section for term breech babies. Most centres (85.0%) indicated that an analysis of relative costs would not affect clinical practice in their setting. CONCLUSION: Clinical practice has changed to planned Caesarean section in most collaborating centres, given the results of the Term Breech Trial. PMID- 12548321 TI - Early ovarian cancer: what is the staging impact of retroperitoneal node sampling? AB - BACKGROUND: Comprehensive surgical staging of apparent early-stage epithelial ovarian cancer includes peritoneal washings, biopsies, and retroperitoneal lymph node dissection. Unresolved is the relative frequency or importance of the lymph node dissection. OBJECTIVES: (1) To determine the site(s) of microscopic metastatic disease in women undergoing a comprehensive staging for apparent early stage cancer of the ovary; (2) to identify those women with metastases in the retroperitoneal lymph nodes alone. METHODS: Between 1985 and 2000, we reviewed all records of women at cancer centres in Winnipeg, Ottawa, and Saskatoon who had undergone a "staging laparotomy" for an apparent early-stage IA epithelial cancer of the ovary. Histology, tumour grade, initial and final surgical stage, and the site(s) of metastatic disease were recorded for all cases. RESULTS: Forty-three of the 128 women (34%) had a final surgical stage of II or III. Sixteen women had positive pelvic biopsies, while 19 had microscopic upper abdominal disease. Eight women had positive retroperitoneal nodes, and in only 2 of these women, disease was found in the retroperitoneal nodes alone. In the 8 women with nodal disease, 5 had grade 3 tumours and 6 had serous histology tumours. CONCLUSION: Comprehensive staging is important to identify women with metastatic disease. Solitary nodal metastases are predominantly found in grade 3 and serous tumours. PMID- 12548322 TI - A randomized controlled trial of outpatient versus inpatient labour induction with vaginal controlled-release prostaglandin-E2: effectiveness and satisfaction. AB - BACKGROUND: Outpatient management in obstetrics is expanding, but evidence to support outpatient labour induction is needed. OBJECTIVE: To compare the effectiveness, acceptability, duration of hospitalization, and safety of outpatient and inpatient induction of labour with intravaginal controlled-release prosta-glandin-E2 (CR-PGE2). METHODS: A prospective, randomized, controlled trial enrolled 300 women at term with parity < or = 5 and singleton pregnancies in cephalic presentation. Each had an unscarred uterus, a normal non-stress test (NST), and a Bishop score of < or = 6. After insertion of the CR-PGE2, and 1 hour of monitoring, those in the outpatient group were discharged home, to return with onset of labour or 12 hours later for an NST. If not already in labour 24 hours later, the women returned for inpatient induction. Vaginal examination was not repeated before 24 hours unless the patient was contracting and required analgesia. Inpatients remained on the antepartum ward but were otherwise treated similarly. The women in both groups reported ratings of satisfaction, pain, and anxiety over the telephone until they were in labour. RESULTS: There were 150 women randomized to outpatient and 150 women to inpatient induction of labour. The number of women who were in labour or who delivered by 24 hours in the outpatient group was 115 (0.77, 95% confidence interval [CI] 0.70-0.84) and in the inpatient group was 107 (0.72, 95% CI 0.64-0.79). The median times to labour were 9.8 hours (95% CI, 8.1-11.4) and 11.4 hours (95% CI, 10.1-12.7), and to delivery were 21.4 hours (95% CI, 19.2-23.5) and 20.7 hours (95% CI, 18.4-23.0), for the outpatient and inpatient groups, respectively. In the outpatient group, 56% of women reported high satisfaction during the initial 12 hours of induction compared to 39% in the inpatient group (p < 0.008). Ratings of pain and anxiety during the first 12 hours of induction were similar. In the outpatient group, women were at home for a median of 8 hours (95% CI, 6.7-9.4) before labour and delivery. There were no significant differences in adverse outcomes. CONCLUSIONS: This study suggests that outpatient induction of labour with intravaginal CR-PGE2 may be a reasonable option for selected low-risk women; however, further study is needed to confirm the safety of this approach. PMID- 12548323 TI - Do environmental contaminants adversely affect human reproductive physiology? AB - There is increasing concern among Canadian women that unwitting and unwanted exposures to environmental contaminants are adversely affecting their health, particularly their ability to become pregnant and have a healthy baby. Evidence of adverse reproductive outcomes among populations exposed to environmental contaminants in the workplace via accidental poisoning, together with detection of environmental contaminant residues in serum and ovarian follicular fluid, has led to the hypothesis that chemical contaminants may be contributing to adverse reproductive outcomes such as infertility, endometriosis, polycystic ovary syndrome, spontaneous abortion, preterm labour, intrauterine growth restriction, and pregnancy-induced hypertension in the general population. The lack of clear evidence concerning the association between exposure to environmental contaminants and adverse reproductive outcomes hampers the clinician's ability to counsel women who are trying to conceive or who have concerns about their pregnancy. This review summarizes the evidence linking environmental contaminant exposure to selected adverse health outcomes by examining the changes in health outcome trends, the consistency of the epidemiological evidence of an association between the health outcome of concern and exposure to environmental contaminants, and the biological plausibility for environmental contaminant mediated effects on human reproductive health. At best, only a moderate association can be found linking exposure to environmental contaminants with evidence of deleterious reproductive effects in women. Lack of disease trend data, weak exposure assessments, and limited mechanistic data supporting the biological plausibility of potential effects are the primary limitations to the hypothesis that exposure to environmental contaminants adversely affects human reproductive physiology. PMID- 12548324 TI - Antenatal corticosteroid therapy for fetal maturation. AB - OBJECTIVES: To assess the benefits and risks of antenatal corticosteroid therapy for fetal maturation. OPTIONS: To administer antenatal corticosteroids or not to women at risk of preterm birth. OUTCOMES: Perinatal morbidity, including: respiratory distress syndrome, intraventricular hemorrhage, infection, adrenal suppression, somatic and brain growth; perinatal mortality; and maternal morbidity, including infection and adrenal suppression. EVIDENCE: MEDLINE and PubMed searches 1996 to August 2002 for English-language articles related to antenatal corticosteroid therapy for fetal maturation, the Cochrane Library, and national statements including that of the National Institutes of Health (NIH), the American College of Obstetricians and Gynecologists, and the Royal College of Obstetricians and Gynaecologists. VALUES: The evidence obtained was reviewed and evaluated by the Maternal-Fetal Medicine Committee of the Society of Obstetricians and Gynaecologists of Canada (SOGC) and recommendations were made according to guidelines developed by the Canadian Task Force on the Periodic Health Exam. BENEFITS AND HARMS: A single course of corticosteroids reduces perinatal mortality, respiratory distress syndrome, and intraventricular hemorrhage. Information regarding repeat courses of corticosteroids is limited and conflicting, with many studies being retrospective and non-randomized. Some studies suggested a reduction in respiratory distress syndrome with repeat courses, but some found increased rates of neonatal and maternal infection; fetal, neonatal, and maternal adrenal suppression; decreased fetal or neonatal somatic and brain growth; and increased perinatal mortality. RECOMMENDATIONS: The SOGC supports the recommendations of the NIH Consensus Development Panel: 1. All pregnant women between 24 and 34 weeks' gestation who are at risk of preterm delivery within 7 days should be considered candidates for antenatal treatment with a single course of corticosteroids. (I-A) 2. Treatment should consist of two 12 mg doses of betamethasone given IM 24 hours apart, or four 6 mg doses of dexamethasone given IM 12 hours apart (I-A). There is no proof of efficacy for any other regimen. 3. Because of insufficient scientific data from randomized clinical trials regarding efficacy and safety, repeat courses of corticosteroids should not be used routinely (II-2E) but be reserved for women participating in randomized controlled trials. VALIDATION: This Committee Opinion has been reviewed and approved by the Maternal-Fetal Medicine Committee of the SOGC and approved by SOGC Council. PMID- 12548325 TI - Novel dermatologic uses of the immune response modifier imiquimod 5% cream. AB - Imiquimod is the first of a new class of drugs to emerge in the treatment of various dermatologic disorders. As an immune response modifier, it has been shown to have potent antiviral and antitumor properties through the stimulation of innate and cell mediated immune pathways. It is currently approved for the treatment of external genital and perianal warts, but has also been found to be an effective treatment for a host of other virus-associated dermatologic lesions, including common and flat warts, molluscum contagiosum and herpes simples 2. Oncological lesions showing improvement with the use of imiquimod include basal cell carcinoma, actinic keratosis, squamous cell carcinoma in situ, malignant melanoma, cutaneous T-cell lymphoma, and cutaneous extramammary Paget's disease. Recent case studies have also found this product to be effective for treating keloids, infantile hemangiomas, porokeratosis of Mibelli, leishmanisis, and tattoo removal. This extensive array of disorders treated successfully with imiquimod warrants further study of this novel and valuable drug. PMID- 12548326 TI - Fat rebalancing: the new "Facelift". AB - The fat compartments of the face undergo changes and are responsible for age related alterations in facial shape. The overlying skin responds to changes in fat in much the same way as a garment clings or sags to the body underneath. By rearranging the underlying fat via suction and fat replacement techniques, a younger-shaped face can be achieved without conventional excisional surgery. PMID- 12548327 TI - Desloratidine for the treatment of chronic urticaria. AB - Chronic urticaria is a common dermatologic condition that is idiopathic in most cases. Antihistamines are the mainstays of treatment for this condition. The newer, second and third generation antihistamines are the preferred agents because of their improved safety profile and comparable efficacy to the first generation antihistamines. Desloratadine is a new non-sedating H1-receptor agonist. Based on clinical studies, desloratadine is a valuable new addition to the available treatment options and should be considered as a first-line therapy for patients with chronic urticaria. PMID- 12548328 TI - Allergic contact dermatitis in children: a practical approach to management. AB - Allergic contact dermatitis (ACD) may account for at least 20% of all childhood dermatitis. Clinically, its morphology is identical to other forms of dermatitis in acute, subacute and chronic forms. A persistent or unusual and localized pattern is often the key to diagnosis. Treatment has centered around the use of corticosteroids, with the adjunct of antihistamines, wet dressings, and emollients for alleviation of symptoms. The newer topical immunosuppressives, tacrolimus and pimecrolimus, may also hold promise as alternative therapies, although they have not been well-studied in this regard. Allergen identification, sometimes through patch testing and allergen avoidance are the keys to preventing recurrences of this disease. PMID- 12548330 TI - Difference in food selection between patas monkeys (Erythrocebus patas) and tantalus monkeys (Cercopithecus aethiops tantalus) in Kala Maloue National Park, Cameroon, in relation to nutrient content. AB - Phytochemical or nutrient analyses of primate diets have revealed clues to their food selection in a single species. On the other hand, few interspecific comparisons of phytochemical or nutrient composition of primate diets have been made, although diets are considered to differ in phytochemical or nutrient content from primate species to species, since different species have different body weights and different morphological and physiological characteristics. I compared the nutrient content of diet between patas monkeys ( Erythrocebus patas) and tantalus monkeys ( Cercopithecus aethiops tantalus) living sympatrically in Cameroon. Patas subsisted on a smaller number of food items, most of which were also tantalus food items. Then, I compared the protein-fiber ratio and the available energy content of the food items eaten by patas (patas foods) with those items eaten only by tantalus (tantalus foods). Both variables were higher in patas than tantalus foods, although there was no significant difference in available energy of plant foods. Next, when I performed discriminant analysis for patas foods and tantalus foods, employing the above two variables, a discriminant function with positive coefficients for both variables was obtained. The mean discriminant-function score of patas foods was higher than that of tantalus foods. Despite being somewhat larger in weight, patas selectively fed on a smaller number of foods of higher quality than did tantalus. I discuss why the results are inconsistent with a well known body weight-diet relationship (Jarman Bell principle). Energy-efficient locomotion enables patas to exploit not only small dispersed food items of high quality but also areas where high-quality foods are distributed in clumps. PMID- 12548331 TI - Positional behavior of free-ranging Japanese macaques (Macaca fuscata). AB - Positional behavior was quantitatively studied in identified free-ranging Japanese macaques (Macaca fuscata). Five male and 11 female adults were observed in a forested mountain habitat. Data were analyzed for proportion of bout distance, number and time of each locomotion and postural type. Japanese macaques are semiterrestrial, and mainly walk and run quadrupedally. This supports the notion that Macaca are generally quadrupeds. Sex differences in positional behavior were found in the preference of substrate and types of positional behavior. Males and females tend to be terrestrial and arboreal, respectively. Males leap more frequently and longer in distance than do females when they are feeding in trees. These sex differences are considered to be related to differences in morphology, food choice, social activity, and the nursing of infants. Frequencies of leaping and the distance covered by leaping in Japanese macaques are more than those of long-tailed macaques which are arboreal quadrupeds. However, Japanese macaques leap shorter distances at a time than do long-tailed macaques, which indicates that body size may be related to leaping distance more than the frequency of leaping and the distance covered by leaping. Japanese macaques are not as specialized for terrestrial locomotion as pig-tailed macaques. They use both terrestrial and arboreal supports, and are considered to be semi-terrestrial quadrupeds, somewhere between the arboreal long-tailed macaque and the terrestrial pig-tailed macaque. PMID- 12548332 TI - Three acoustic forms of Allen's galagos (primates; Galagonidae) in the Central African region. AB - This study identifies populations currently classified as Allen's galago (Galago alleni) at ten locations in Gabon, Cameroon and Bioko Island. Morphological diversity was evident both within and between populations. Attention to the loud calls revealed three distinct vocal profiles which are consistent within biogeographical regions. This work is based on the Recognition Concept of Species which refers to a Specific Mate Recognition System. Galagos rely less on visual signals than diurnal primates and recognise each other principally by means of auditory and olfactory signals. Galagos possess repertoires of loud calls relating to contact and alarm which are thought to be species-specific. Other studies of nocturnal prosimians (galagos, tarsiers) have demonstrated that the unique loud call repertoires are reliable indicators of species boundaries; whereas characters such as body size and pelage coloration are highly variable, even within populations. The vocal data in this study provide evidence of at least three acoustic forms of galago within the Allen's group which are predicted to represent three distinct species: the Allen's form on Bioko Island and south west Cameroon, the Gabon form in southern Cameroon and northern Gabon and the Makande form in Gabon south of the Ogooue river. Some populations may be vulnerable to extinction due to limited distributions and habitat destruction. PMID- 12548333 TI - Comparative morphology of the hyo-laryngeal complex in anthropoids: two steps in the evolution of the descent of the larynx. AB - The descent of the larynx is a key phenomenon not only in postnatal development, but also in the evolution of human speech. The positional change of the larynx is affected by the descent of the hyoid bone in relation to the mandible and cranial base, and that of the laryngeal framework in relation to the hyoid bone. The phylogeny of the spatial configuration of the hyo-laryngeal complex is one of the most important sources of information for elucidating the evolution of laryngeal descent. In the present study, the anatomy of the complex was examined in various species of anthropoids to compare the configuration, the shape of the basihyal and thyroid cartilage, and the length of the lateral thyrohyoid ligaments. Non human hominoids share most features with humans, while cercopithecoids and ceboids have anatomical features that sharply contrast to humans, except for the form of the thyroid cartilage in ceboids. The laryngeal framework in hominoids is well separated from and assured of mobility independent of the hyoid. In cercopithecoids and ceboids, it is, by contrast, locked into and tied tightly with the hyoid so that the hyo-laryngeal complex acts as a functional unit. This spatial configuration is considered to be significantly related to the mechanism that prevents aspiration, including epiglottic movement and vestibular closure. Non-human hominoids are inferred to share the mechanism with human adults, not with cercopithecoids and ceboids, although their larynx is located as high as the latter. Consequently, it is hypothesized that the descent of the larynx evolved in two steps. The first step would have been descent of the thyroid in relation to the hyoid for the evolution of the mechanism preventing aspiration, which occurred in the common ancestor of hominoids. The second step, descent of the hyoid within the neck, occurred during hominid evolution for human speech. PMID- 12548334 TI - Altitudinal and seasonal variations in the diet of Japanese macaques in Yakushima. AB - Altitudinal and seasonal variations in the diet of Japanese macaques in Yakushima, southwestern Japan, were studied for 2 years by means of fecal analysis. The altitudinal range of fecal samples collected was 30 m to 1,203 m above sea level, and it was divided into three zones: low-zone forest (0-399 m), middle-zone forest (400-799 m), and high-zone forest (800 m-1,230 m). There was a considerable altitudinal and seasonal variation in the macaques' diet. Seed/fruit and animal matter were eaten more in the lower zones, whereas more fiber and fungi were consumed in the higher zones. In all of the zones, they ate seed/fruits the most in autumn (September-November) and the least in spring (March-April). They ate fibrous food the most in spring and the least in autumn. Macaques relied on seed/fruits heavily in the lower zone for a longer period than in the higher zones. Macaques in the high-zone forest ate almost no seed/fruit foods from March to May. Altitudinal variations in availability of seed/fruit foods seem to have influenced the altitudinal variations in diet. Total basal area of seed/fruit-food trees, species richness of seed/fruit-foods, main seed/fruit-food types available, and annual fleshy-fruit production all decreased with increasing altitude. Both interannual variation and annual cyclicity of diet were found in all zones. PMID- 12548335 TI - Increase in tannin consumption by sifaka ( Propithecus verreauxi verreauxi) females during the birth season: a case for self-medication in prosimians? AB - In this study we report preliminary data on the consumption of tannin-rich plants by sifakas (Propithecus verreauxi verreauxi) living in the Kirindy forest, western Madagascar. Sifakas spent most of their time feeding on only a few plant species. The tannin intake during the period between the pregnancy and birth season was significantly higher in pregnant females or females with lactating infants than in non-reproductive females and males. These periparturient females secured a larger proportion of condensed tannins by short feeding bouts on plants not included in the group's limited preferred food species. The measured increase in tannin intake is puzzling in light of the fact that tannins are commonly known for their protein-binding properties. Since protein demands are highest in pregnant and lactating females, possible medicinal benefits of tannin ingestion are considered. Tannin consumption is associated with an increase in body weight and stimulation of milk secretion. Veterinarians administer tannins as an astringent, anti-hemorrhagic and anti-abortive. Their high potential as an alternative anthelminthic has also recently been recognized. Thus, when viewed as self-medicating behavior, controlled increase in tannin intake could have multiple prophylactic advantages for females during the periparturient period. The high selectivity in their plant choice, and the presence of unusual feeding habits by a particular group of individuals (females with infants) limited in time (birth season), suggests that an increase in tannin ingestion may be a self medicating behavior with multiple directly adaptive benefits to female reproduction. PMID- 12548336 TI - Successful reproduction in wild-released orphan chimpanzees (Pan troglodytes troglodytes). AB - We report a case of successful reproduction in wild-released orphan chimpanzees. Using non-invasive genetic analysis, we determined the paternity of an infant born to a female chimpanzee released by HELP (Habitat Ecologique et Liberte des Primates) Congo into the Conkouati-Douli National Park, Republic of Congo. The sire was a released male, thus demonstrating successful reproduction in both male and female released chimpanzees. These results provide evidence that release into the wild may be a viable response to the plight of orphan chimpanzees in Africa, and we discuss further applications of non-invasive genetic tagging to release programs. PMID- 12548337 TI - Long distance mobility of male Japanese macaques evidenced by mitochondrial DNA. AB - A 487-nucleotide sequence in the polymorphic D-loop region of matrilineally inherited mitochondrial DNA was compared in samples from 14 (9 adult males and 5 females or young) Japanese macaques (Macaca fuscata fuscata) of two troops at Gagyusan in Takahashi City, Okayama Prefecture, Japan. The majority (7 out of the sampled 9) of the adult males associated with the troops had nucleotide sequences of six distinct types, all of which were different from that of the Gagyusan troop females, whereas two young solitary males shared the same sequence. The present study exemplified a case in which the majority of adult males immigrated from outside the local population. Notably, two of the males were revealed to share the same sequence determined for a matriline 45 km away. PMID- 12548338 TI - Common daily activities in the virtual environment: a preliminary study in parkinsonian patients. AB - Patients with Parkinson's disease (PD) create behavioral motor strategies by using external cues to facilitate their movements. Virtual reality (VR) could work as an external stimulus in order to explore the motor plans by means of creation of mental images. We tested 2 women with PD aged 68 and 69 years, and 10 normal control subjects. Patients underwent a neuropsychological assessment to evaluate cognitive abilities involved in the tasks required by the VR session. VR environment reproduces common daily activities situations at home, such as eating or using the bathroom. VR describes the alterations of the motor plans in PD by a point of view different from the clinical one, by testing "pure" mental sequences of the execution of a movement, without the interference of motor disability. PMID- 12548339 TI - Striatal dopaminergic denervation in early and late onset Parkinson's disease assessed by PET and the tracer [11C]FECIT: preliminary findings in one patient with autosomal recessive parkinsonism (Park2). AB - Neuroimaging studies of striatal dopamine transporters (DAT) have shown that this measurement is a specific marker of dopaminergic degeneration in patients with Parkinson's disease. However, little data is available in subjects with early disease onset, particularly in those with autosomal recessive parkinsonism. We measured striatal DAT binding in 10 patients with early onset PD (onset <40 years) and in 10 with late onset PD (onset >50 years) using PET and the tracer [(11)C]FECIT. One early onset subject presented a mutation in the parkin gene consistent with autosomal recessive parkinsonism. Data were compared with those of 15 control subjects. We found a comparable decrement of striatal DAT binding in early and late onset PD. Loss was widespread and bilateral in the patient carrying the Park2 mutation, suggesting a different pattern of denervation in these individuals. PMID- 12548340 TI - Body weight, levodopa pharmacokinetics and dyskinesia in Parkinson's disease. AB - We conducted a pharmacokinetic study in 164 patients with sporadic Parkinson's disease (PD) to address the relationship between body weight and levodopa pharmacokinetics. Patients underwent an oral acute levodopa test with 250 mg levodopa and pharmacokinetic variables were further assessed. Plasmatic levodopa area under the curve (AUC-l) and body weight were significantly and inversely correlated. Women were significantly lighter and more dyskinetic than men, and had greater AUC-l values. Our data suggest that during long-term treatment, lighter PD patients, especially women, may receive a greater cumulative dosage of levodopa per kilogram of body weight. This could explain gender differences for the development of levodopa-induced peak-dose dyskinesias observed during the course of the disease. PMID- 12548341 TI - Serotonin syndrome: a reported case. AB - We describe a patient treated with SSRI and Ldopa, who developed agitation, rigidity, hyperreflexia, restlessness, autonomic instability, fever and finally death. CSF examination, MRI of the brain, laboratory investigations, except for serum CK, glycemia and WBC, were normal. His condition was thought to result from an central serotonin activity. The serotonin syndrome occurs following the use of serotomimetic agents (serotonin reuptake inhibitors, tricyclic and tetracyclic antidepressants, tryptophan alone or in combination with monoamine oxidase inhibitors). PMID- 12548342 TI - Effects of the intrastriatal administration of selective dopaminergic agonists on Fos expression in the rat brain. AB - In this study, we mapped the cerebral expression of Fos protein following intrastriatal stimulation of D(1) or D(2) receptors, in freely moving animals. Animals treated with the D(1) agonist SKF 38393 showed massive Fos increases in the cerebral cortex, ipsilaterally to the injected striatum, which were counteracted by systemic administration of D(1) antagonist SCH 23390. Conversely, D(2) agonist quinpirole suppressed cortical expression of Fos, while systemic administration of D(2) antagonist eticlopride relieved this blockade. As for the basal ganglia, Fos was consistently expressed only in the injected striatum of rats receiving SKF 38393. These results show that striatal dopamine receptors may play a role in the modulation of cortical activity. They also provide new information on a class of drugs--the dopamine agonists--whose role in the therapeutic strategy of Parkinson's disease is continuously evolving. PMID- 12548343 TI - Autosomal recessive early onset parkinsonism is linked to three loci: PARK2, PARK6, and PARK7. AB - Autosomal recessive, early onset parkinsonism (AREP) is genetically heterogeneous. Mutations in the parkin gene (PARK2 locus, chromosome 6q) account for up to 50% of AREP families. The parkin protein displays ubiquitin-ligase activity for different targets, which accumulate in the brain of patients with parkin defect and might cause neurodegeneration. Two new AREP loci (PARK6 and PARK7) have been recently mapped on chromosome 1p and confirmed in independent datasets, suggesting that both might be frequent. The three AREP forms display similar clinical phenotypes. Recruiting new families will help cloning the defective genes at PARK6 and PARK7 loci. This will contribute to unraveling the pathogenesis of AREP, and it is also expected to foster our understanding of molecular events underlying classic Parkinson's disease. PMID- 12548344 TI - Perfusion-weighted dynamic susceptibility (DSC) MRI: basal ganglia hemodynamic changes after apomorphine in Parkinson's disease. AB - Relative regional blood flow of basal ganglia was studied by means of perfusion weighted dynamic susceptibility (DSC) MRI. Parkinson's disease (PD) patients showed a significant inter-hemispheric asymmetry due to a higher perfusion in the more affected side, while normal subjects did not. PD exhibited an abnormal "asymmetry index" in the measured nuclei. A second DSC-MRI examination performed after subcutaneous apomorphine administration did not show any significant asymmetry in PD patients. DSC-MRI of basal ganglia confirms the asymmetry observed in PET studies of PD patients, suggesting that this method is a promising and low-cost technique in neurodegenerative diseases. PMID- 12548345 TI - Frequency, distribution and characteristics of progressive supranuclear palsy in Italy: preliminary observations. AB - We report the number of pathologically proven cases of progressive supranuclear palsy, described in the Italian neurological literature from 1961 until now. A discussion of the diagnostic value of downward gaze palsy is made. A comparison with the number of similar cases described in the rest of the world and with the number of Parkinsonian patients who died in the same region in the corresponding year is attempted. PMID- 12548346 TI - Frontal intermittent rhythmic delta activity (FIRDA) in patients with dementia with Lewy bodies: a diagnostic tool? AB - The accuracy of the clinical diagnosis of dementia with Lewy bodies (DLB) remains poor, especially in early phases of the disease, in spite of applying current consensus diagnostic criteria. The need for supportive diagnostic tools is therefore warranted. In this study EEG recordings showed a main pattern of bilateral frontal intermittent rhythmic delta activity (FIRDA) in 7 of 10 patients, aged 58-83 years, 8 of whom were diagnosed as affected by "probable" and 2 by "possible" DLB. Conversely, the same EEG abnormality was found only in 2 of 9 age-matched patients, 8 of whom had "probable" and 1 "possible" Alzheimer's disease, according to NINCDS-ADRDA criteria, taken as controls. The degree of cognitive impairment was comparable among the two groups of patients. If these findings will be confirmed in a larger series, FIRDA, even though an aspecific EEG pattern, could be of value in improving the diagnostic accuracy of DLB. PMID- 12548347 TI - Bipolar affective disorder and Parkinson's disease: a rare, insidious and often unrecognized association. AB - Five patients (4 women) with Parkinson's disease (PD) and primary major psychiatric disorder (PMPD) meeting DSM-IV criteria for the diagnosis of bipolar affective disorder (BAD) were studied. Four patients had early onset PD. Four developed a severe psychiatric disorder a few years after starting dopaminergic therapy in presence of a mild motor disability and a mild cognitive impairment, with no evidence of cerebral atrophy at CT or MRI. Two patients developed a clear manic episode; the other three presented a severe depressive episode (in one case featuring a Cotard syndrome). None showed previous signs of long term L-dopa treatment syndrome (LTS), hallucinosis or other minor psychiatric disorders. The two manic episodes occurred shortly after an increase of dopaminergic therapy and in one case rapid cyclic mood fluctuations were observed. At the onset of psychiatric symptoms, all patients had an unspecific diagnosis of chronic delusional hallucinatory psychosis (CDHP). PMID- 12548348 TI - Modification of respiratory function parameters in patients with severe Parkinson's disease. AB - Respiratory dysfunction remains one of the most common causes of death in patients with complicated Parkinson's disease (PD). The aim of this study was to investigate pulmonary function in fluctuating PD patients during "on" and "off" states of the disease. We studied 12 fluctuating, non-smoking PD patients (H&Y stages 3-5) without a history of lung or cardiovascular disease; all patients underwent Hoehn and Yahr scale (H&Y) and Unified Parkinson Disease Rating Scale (UPDRS items 18-31) to evaluate extrapyramidal impairment, as well as pulmonary function tests (PFT) and arterial blood gas analyses to assess respiratory function. All evaluations were performed during a stable on state of disease and in an off state produced by 12 hours of therapy withdrawal. A restrictive pattern of flow-volume loop was observed both in on and off states of disease. In the off state, we found a significant worsening in both FEV1 and FVC; the FEV1/FVC ratio was unmodified. These results suggest a restrictive pattern of flow-volume loop in these patients. PMID- 12548349 TI - Visualisation of the subthalamic nucleus: a multiple sequential image fusion (MuSIF) technique for direct stereotaxic localisation and postoperative control. AB - A novel multiple, sequential image fusion (MuSIF) procedure merging stereotaxic CT with frameless magnetic resonance imaging (MRI) is used since June 2000 to visualise and directly localise the subthalamic nucleus (STN) on T2 images. In 13 consecutive Parkinson's cases, intraoperative recording and stimulation verified bilateral electrode implantation guided by fused T2 images. In 85% of sides, final implantation opted for visualised target track. Implanted electrode position on postoperative T2 images matched planned target. Clinical follow-up reproduces literature's best results. This MuSIF technique, effective for direct STN targeting, has practical advantages: MRI can be performed regardless of surgery time; regular MR scanning to correct real image distortion is unneeded; and the need for multiple localising tracks is reduced by enabling us to account for each patient's STN anatomy. PMID- 12548350 TI - Psychophysiological approach in Parkinson's disease: L-dopa effects on preprogramming and control activity. AB - To investigate whether preprogramming (Bereitschaftspotential, BP) and control activity (skilled performance positivity, SPP) in a complex task are sensitive to L-dopa, movement related potentials (MRPs) were recorded in 12 non-demented Parkinson's disease (PD) patients before and after acute L-dopa administration, and in 17 control subjects. After L-dopa administration, the PD patients scored a significantly higher percentage of correct performances ( p<0.05), linked to a decreased BP amplitude ( p<0.001) and an increased SPP amplitude ( p<0.005), than before therapy. Our findings suggest that preprogramming activity is impaired in untreated PD patients. Dopaminergic drug administration seems to restore their ability to use more automatic motor strategies which become more similar to that of normal subjects. PMID- 12548351 TI - Noradrenergic loss enhances MDMA toxicity and induces ubiquitin-positive striatal whorls. AB - Movement disorders involve a number of neurodegenerative conditions, mostly affecting basal ganglia. Parkinson's disease (PD) is classically defined by the selective loss of dopaminergic neurons in the substantia nigra pars compacta. Administration of specific neurotoxins represents a common tool to reproduce this lesion. Among these, amphetamine derivatives act as powerful monoamine neurotoxins, impairing striatal dopamine (DA) axons in mice. Despite the well investigated effects on striatal DA terminals, only sporadic studies have focused on the potential toxicity of amphetamines towards post-synaptic neurons within the striatum. In the present work we found that 3,4-methylenedioxymethamphetamine (MDMA) produces ultrastructural alterations in striatal cells, featuring as membraneous whorls, positive for ubiquitin and heat shock protein 70. These morphological alterations were enhanced in locus coeruleus-lesioned mice. PMID- 12548352 TI - Clinical predictors in Parkinson's disease. AB - Parkinson's disease is characterized by heterogeneity of clinical presentations, association of signs and symptoms, rate of progression, and response to therapy. The aim of this prospective 5-year study was to evaluate whether clinical features at onset were predictive of the subsequent progression. Two courses were identified which differed in the characteristics at onset. Slow course was characterized by earlier age at onset, lateralization of motor signs, rest tremor, and absence of gait disturbance. Rapid course presented older age, less evident lateralization of signs, predominance of bradykinesia-rigidity and gait disturbance. Our results confirmed that PD is clinically heterogeneous and specific patterns of onset seem to be associated with different rates of disease progression. Predictive models based on these clinical characteristics have a good sensitivity in indicating a slow disease progression but are not reliable in indicating a rapid evolution. PMID- 12548353 TI - Screening cognitive decline in dementia: preliminary data on the Italian version of the IQCODE. AB - The IQCODE is a retrospective questionnaire for caregivers about changes which occurred in a patient's cognitive and functional efficiency in the previous 10 years of life. Previous studies demonstrated the validity of the IQCODE for the screening of dementia similar to that of traditional cognitive screening tests, with the additional advantage of allowing the detection of cognitive change, rather than just cognitive impairment. The present paper deals with the preliminary results of the validation of the Italian version of the questionnaire in a sample of 45 mild to severely demented patients and 13 patients with mild cognitive impairment (MCI), compared to 20 cognitively intact elderly subjects. The IQCODE demonstrated satisfactory discriminative power for dementia as well as for MCI and a good correlation with the MMSE. PMID- 12548354 TI - Frequency of apraxia of eyelid opening in the general population and in patients with extrapyramidal disorders. AB - We ascertained the prevalence of apraxia of eyelid opening (AEO) in a community located in Puglia, a region of southern Italy. The crude prevalence rate was 59 per million (95% confidence interval, 24-173). AEO coexisted with adult onset blepharospasm in 75% of cases, with atypical parkinsonism in 25% of cases. Among the overall patient population seen at our movement disorders clinic from 1987 to 1997, AEO was isolated in 10 otherwise healthy individuals, associated with adult onset dystonia in 13 cases, and associated with a parkinsonian syndrome in 9 cases. The frequency of AEO was 10.8% in the dystonia group, and 2.1% in the overall parkinsonian group (Parkinson's disease, 0.7%; progressive supranuclear palsy, 33.3%). In two patients with possible progressive supranuclear palsy, AEO worsened after increasing levodopa dosage or acute apomorphine challenge and disappeared following levodopa discontinuation. AEO developing in the setting of a parkinsonian syndrome may be either disease- or drug-related. PMID- 12548355 TI - Amantadine in Huntington's disease: open-label video-blinded study. AB - Huntington's disease (HD) is characterized by chorea, cognitive and behavioral changes. Amantadine, a non-competitive NMDA receptor antagonist, has shown an antidyskinetic effect on levodopa-induced dyskinesias, which are known to have strict pathogenetic analogies with choreic hyperkinesias. The antidyskinetic efficacy of amantadine and its effects on cognitive and behavioural symptoms were evaluated. Eight HD patients received oral amantadine (100 mg tid) unblinded for a 1-year period. A significant reduction of dyskinesias was reported ( p<0.01). No changes were observed in neuropsychologic and psychiatric assessments after 6 and 12 months of therapy. These data may have relevance to the treatment of HD with amantadine. PMID- 12548356 TI - Parkinson's disease and reproductive life events. AB - Onset, progression and duration of Parkinson's disease (PD) seem to be similar in men and women but gender differences have been suggested concerning clinical aspects, such as more severe disease in men and more dyskinesia in women. Taking into account the multiple influences of sex hormones, estrogens in particular, on basal ganglia function, the present work compared the characteristics of reproductive events in PD subjects and in healthy women, with regard to onset and clinical aspects of the disease with respect to the milestones of reproductive life. A total of 150 PD women and 200 healthy women matched for age were interviewed about reproductive life and disease characteristics (if patients). As a group, the women with PD had menarche later than the controls, but in the normal range. Menopause was similar to the controls for time, type (natural) and onset (slow), but with less hormonal therapies. Women with PD had fewer children, while breast feeding and gynecological diseases were comparable to controls. The characteristics of menses were similar as far as dysmenorrhea and premenstrual syndrome (PMS). The women with PD onset before menopause had a longer disease duration, with a more frequent fluctuating stage, and longer treatment with both levodopa and dopamine agonists. They had more dysmenorrhea and PMS when compared with women with PD onset after menopause and controls. PMID- 12548357 TI - Magnetic resonance relaxometry in Parkinson's disease. AB - A central role of iron in the pathogenesis of idiopathic Parkinson's disease (PD), due to its increase in substantia nigra pars compacta dopaminergic neurons and its capacity to enhance production of toxin reactive oxygen radicals, has been discussed for many years. Nuclear magnetic resonance (NMR) relaxation is considered an objective and noninvasive method of measuring regional iron concentrations. By means of this technique we investigated both controls and PD patients. PMID- 12548358 TI - Quetiapine versus clozapine: a preliminary report of comparative effects on dopaminergic psychosis in patients with Parkinson's disease. AB - This study investigated the efficacy and safety of quetiapine versus clozapine in parkinsonian patients with dopaminergic psychosis. All patients fulfilling the inclusion criteria were randomly assigned to receive either quetiapine or clozapine. The duration of the trial was 12 weeks. The severity of psychosis was assessed using the BPRS and the Clinical Global Impression Scale-Severity subscale (CGI-S). The UPDRS III was used to monitor the progression of PD during the study period. Twenty patients, 10 on clozapine, and 10 on quetiapine, completed the study. The psychopathological state, as assessed by the BPRS and by the CGI-S, improved significantly ( p<0.001) from baseline in both treatment groups. No differences were found between clozapine and quetiapine at each assessment time. The UPDRS score decreased significantly ( p<0.05) in the clozapine group, while was almost unchanged in the quetiapine group. PMID- 12548359 TI - Incidence of RBD and hallucination in patients affected by Parkinson's disease: 8 year follow-up. AB - We describe the 8-years follow-up of 80 patients affected by idiopathic, L-dopa responsive Parkinson's disease. All patients were evaluated at baseline and during the follow-up with visual evoked potential, P300 event related potentials and polysomnography. The patients and their relatives compiled sleep and hallucination questionnaires. Statistical analysis was performed to evaluate if visual abnormalities, abnormal P300 recordings or sleep disturbances were linked to the development and hallucinations. Our results show that abnormal vision and abnormal P300 did not correlate with the incidence of hallucinations. However, the presence of REM sleep behavioral disorder (RBD) was significantly related to the development of hallucinations,independently of age, gender or duration of disease but dependent on the amount of dopaminoagonist treatment. PMID- 12548360 TI - Cerebrospinal fluid levels of biomarkers and activity of acetylcholinesterase (AChE) and butyrylcholinesterase in AD patients before and after treatment with different AChE inhibitors. AB - In order to evaluate the biochemical effects of long-term treatment with inhibitors of acetylcholinesterase (AChE) in patients with Alzheimer's disease (AD), we measured the activities of AChE and butyrylcholinesterase (BuChe) and the concentrations of beta-amyloid (1-42), tau and phosphorylated tau proteins in the cerebrospinal fluid (CSF). A total of 91 patients suffering from probable AD of mild to moderate degree were treated for 6 months with donepezil (n=59), galantamine (n=15), rivastigmine (n=10), or placebo (n=7). AChE activity in CSF was significantly increased after treatment with donepezil and galantamine; the opposite was observed in the rivastigmine-treated group. Untreated patients did not show any AChE activity variation. BuChE did not show any change in any of the groups studied. Mean values of beta-amyloid(1-42), total tau and phosphorylated tau also did not vary significantly. We conclude that AChE inhibitors induce different effects on CSF AChE activity, while other CSF biomarkers are not significantly affected by treatment. PMID- 12548361 TI - Cytogenetic alterations in lymphocytes of Alzheimer's disease and Parkinson's disease patients. AB - We investigated the presence of cytogenetic alterations in peripheral blood lymphocytes of Alzheimer's disease (AD) and Parkinson's disease (PD) patients. Detection of spontaneous structural and/or numerical chromosome damage has been assessed by micronucleus (MN) assay coupled with fluorescence in situ hybridization (FISH). The cytogenetic investigation was performed on 22 AD patients, 18 PD patients, and 20 controls. The spontaneous frequencies of micronuclei (MN) in human lymphocytes of both AD and PD patients were significantly higher than in controls. The majority of MN was composed of whole chromosomes in AD patients, while a prevalence of MN arising from chromosome breakage was observed in PD patients. Different molecular mechanisms underlie cytogenetic alterations observed in peripheral lymphocytes of AD and PD patients. PMID- 12548362 TI - Controlled-release transdermal apomorphine treatment for motor fluctuations in Parkinson's disease. AB - This study evaluated the efficacy in Parkinson's disease (PD) of a new pharmacologic preparation of apomorphine included in microemulsions and administered by transdermal route, which provides a constant release of the drug for several hours (Apo-TD). Twenty-one PD patients with motor fluctuations were treated with L-dopa alone, with L-dopa plus oral dopamine-agonists, or with L dopa plus Apo-TD. Apo-TD improved UPDRS-III and tapping test scores in "off" conditions, and reduced duration of "off" periods; no improvement in "on" conditions occurred. We conclude that Apo-TD shows its efficacy particularly by reducing "off" period duration and disability rather than improving motor performances in "on" conditions and therefore it seems a promising treatment for uncontrolled "off" phases in PD patients. PMID- 12548363 TI - Movement-related modulation of neural activity in human basal ganglia and its L DOPA dependency: recordings from deep brain stimulation electrodes in patients with Parkinson's disease. AB - Through electrodes implanted for deep brain stimulation in three patients (5 sides) with Parkinson's disease, we recorded the electrical activity from the human basal ganglia before, during and after voluntary contralateral finger movements, before and after L-DOPA. We analysed the movement-related spectral changes in the electroencephalographic signal from the subthalamic nucleus (STN) and from the internal globus pallidus (GPi). Before, during and after voluntary movements, signals arising from the human basal ganglia contained two main frequencies: a high beta (around 26 Hz), and a low beta (around 18 Hz). The high beta (around 26 Hz) power decreased in the STN and GPi, whereas the low beta (around 18 Hz) power decrease was consistently found only in the GPi. Both frequencies changed their power with a specific temporal modulation related to the different movement phases. L-DOPA specifically and selectively influenced the spectral power changes in these two signal bands. PMID- 12548364 TI - High-frequency electrical stimulation of the subthalamic nucleus in Parkinson's disease: kinetic and kinematic gait analysis. AB - In the advanced phase of Parkinson's disease (PD), gait disturbances represent one of the main causes of disability. Several studies demonstrated that high frequency electrical stimulation (HFS) of the subthalamic nucleus (STN) significantly improves the motor symptoms of PD. This study was finalised to quantitatively analyze the effect of STN HFS on gait of PD patients, through a three-dimensional gait analysis system. Ten PD patients were studied, with and without STN HFS. The results demonstrated that STN HFS significantly improves all the main gait parameters in PD patients. PMID- 12548365 TI - Cognitive and psychiatric characterization of patients with Huntington's disease and their at-risk relatives. AB - We examined cognitive and psychiatric disturbances in patients with Huntington's disease (HD) in comparison to at-risk asymptomatic subjects. Cognitive and psychiatric scales and an HD motor scale were administered to 40 HD patients, 17 pre-symptomatic HD gene carriers (AR+) and 28 non gene carriers (AR-). HD patients did worse than AR+ and AR- in all motor, cognitive and psychiatric measures, while AR+ and AR-subjects did not differ between each other. HD patients had high scores for negative psychiatric symptoms, but there was no correlation between illness duration and psychiatric or cognitive performance. In HD, disease course and symptomatology are heterogeneous and negative psychiatric symptoms are common. PMID- 12548366 TI - CAG mutation effect on rate of progression in Huntington's disease. AB - Huntington's disease (HD) is progressively invalidating and caused by a CAG expanded mutation. We tested the effect of the mutation length on the rate of progression in a cohort of 80 patients clinically followed-up and genetically characterized. Two patients presenting an infantile and aggressive HD form starting under 10 years had over 90 repeats; the other patients did not show any influence of the CAG expanded number on the rate of progression. In conclusion, the CAG expanded repeat affects the disease progression only at a very upper pathological range and in rare cases initiating very early in the life, while it does not seem to affect in any way the severity of the phenotype in most HD patients. Other factors affecting the motor symptom progression, other than the expanded repeats, therefore have to be investigated. PMID- 12548367 TI - Anatomo-clinical correlation of intraoperative stimulation-induced side-effects during HF-DBS of the subthalamic nucleus. AB - The efficacy of deep brain stimulation of the subthalamic nucleus (STN) is dependent on the accuracy of targeting. In order to reduce the number of passes and, consequently, the duration of surgery and risk of bleeding, we have set up a new method based on direct magnetic resonance imaging (MRI) localisation of the STN. This procedure allows a short duration of the neurophysiological session (one or two initial tracks). Whenever a supplementary track is needed, the stimulation-induced side effects are analysed to choose from one of the remaining holes in Ben's gun. A good knowledge of anatomical structures surrounding the STN is mandatory to relate side effects to the actual position of the track. In our series of 11 patients (22 sides, 37 tracks), the most common and reproducible side effects were those characterised by motor, sensorial, oculomotor and vegetative signs and symptoms. Moreover, the therapeutic window (distance between the current intensity needed to obtain the best clinical effect and the intensity capable to induce side effects) predicted clinical efficacy in the long-term, and contributed to the choice of which among the examined tracks had to be implanted with the chronic macroelectrode. PMID- 12548368 TI - Deep brain stimulation of the subthalamic nucleus in Parkinson's disease: long term follow-up. AB - Deep brain stimulation (DBS) of the subthalamic nucleus (STN) has been shown to be an effective therapy for the treatment of advanced Parkinson's disease (PD). Forty-seven patients were bilaterally implanted for STN DBS and clinically evaluated according to the Core Assessment Program for Intracerebral Transplantations before surgery and 3, 12 and 24 months after surgery. Electrical stimulation led to a significant improvement in motor symptoms and in the quality of life, allowing a significant reduction of dopaminergic drugs with a consequent improvement of drug-induced dyskinesias. Statistical differences were observed between UPDRS parts II, III and IV values and daily levodopa dosage in the pre- and postoperative periods, while no differences were evident between the 3 postoperative conditions. PMID- 12548369 TI - The role of somatosensory feedback in dystonia: a psychophysical [correction of psycophysical] evaluation. AB - Ten patients with idiopathic dystonia and twelve healthy controls were tested with pairs of non-noxious electrical stimuli separated by different time intervals. Stimuli were delivered (i) to the pad of the index finger (same-point condition), (ii) to the pad and to the base of the index finger (same-finger condition) and (iii) to the pad of the index and ring fingers (different-fingers condition). Subjects were asked to report if they perceived single or double stimuli in the first condition and synchronous or asynchronous stimuli in the second and third conditions. STDTs were significantly higher in dystonic than control subjects in all three conditions. Results extend current knowledge on deficits of somesthetic temporal discrimination in dystonia by showing that temporal deficits are not influenced by spatial variables. PMID- 12548370 TI - Combination of two different dopamine agonists in the management of Parkinson's disease. AB - An alternative approach to the symptomatic treatment of parkinsonian patients with and without motor fluctuations is to use dual dopamine agonists. The aim of this study was to investigate the symptomatic effect of administrating a second dopamine agonist to parkinsonian patients already assuming pramipexole or ropinirole. As the second dopamine agonist we chose cabergoline, a drug with a long half life, whose pharmacological profile differs from that of the newer non ergot-derived dopamine-receptor agonists. In this pilot study we enrolled 27 patients: 21 patients had motor fluctuations and were receiving levodopa plus a dopamine agonist, and 6 patients without motor fluctuations were receiving a dopamine agonist without levodopa. This open study shows that dual dopamine agonist therapy (cabergoline plus pramipexole or ropinirole) may be used in the symptomatic treatment of patients with Parkinson's disease receiving therapy with or without levodopa. PMID- 12548371 TI - PARK6 is a common cause of familial parkinsonism. AB - The Parkin gene is responsible for about 50% of autosomal recessive juvenile parkinsonism (ARJP) and less than 20% of sporadic early onset cases. We recently mapped a novel ARJP locus (PARK6) on chromosome 1p. Linkage to PARK6 was confirmed in 8 families from 4 different European countries. These families share some clinical features with the European Parkin-positive cases, with a wide range of ages at onset and slow progression. However, features typical of ARJP, such as dystonia and sleep benefit, were not observed, making the clinical presentation of late-onset cases indistinguishable from that of idiopathic PD. The determination of the smallest region of homozygosity in one consanguineous family allowed reducing the candidate interval to 9 cM. PARK6 appears to be an important locus for ARJP in Europe. PMID- 12548372 TI - Evaluation of risk of Parkinson's disease in a cohort of licensed pesticide users. AB - In the last two years, the environmental theory on the aetiology of Parkinson disease has acquired new data. From an experimental point of view, a new model of parkinsonism induced by rotenone, a diffuse insecticide, has been proposed, and in vitro studies have provided proof that several pesticides stimulate the formation of alpha-synuclein fibrils (one of the principal constituents of Lewy bodies). Moreover, a meta-analysis of all case-control studies so far performed showed a positive, statistically significant association between pesticide exposure and PD. In this context, we are performing a cohort study on 5575 licensed pesticide users in the province of Viterbo. After 27 years of follow-up, 4788 subjects are still alive. The aim of this study is to measure the prevalence of Parkinson's disease in a large group of workers with theoretically increased risk. PMID- 12548373 TI - Clinical assessment of dysphagia in early phases of Parkinson's disease. AB - Dysphagia is a frequent symptom in parkinsonism, but it is less commonly reported by patients with idiopathic Parkinson's disease (IPD), especially in the early phases. Sixty-five patients with IPD were questioned about symptoms of dysphagia and an objective swallowing test was administered. Reduced swallowing speed for food and complaints of food sticking in the throat, wet voice and cough after liquid intake and nocturnal sialorrhea were reported, respectively, by 35%, 20% and 15% of patients. On objective examination, oral-phase (facial, tongue and palatal musculature) abnormalities were found in 70% of patients. Lingual transfer movements, mainly propulsion, and palatal elevation were severely hypokinetic. Wet voice after liquid intake and cough reflex after solid/liquid intake were detected in 40% of patients. On the other hand, severe dysphagia with frequent food aspiration and chest infections requiring antibiotics in the last 12 months was not found; cough reflex was retained in all patients. On the basis of these results, a regular assessment on swallowing abilities in patients with IPD is warranted in the clinical setting because with simple dietary advice and a short rehabilitative training, the quality of life in these patients can be improved. PMID- 12548374 TI - ALS-plus: 5 cases of concomitant amyotrophic lateral sclerosis and parkinsonism. AB - According to El Escorial criteria, amyotrophic lateral sclerosis (ALS), combined with other neurologic disorders, such as dementia and parkinsonism, is defined as ALS-plus. These overlaping syndromes are extremely rare. Here we report 5 cases (3 men, 2 women) of ALS-plus; mean age at the onset of symptoms was 67 years (range, 65-72). In 3 patients, motoneuronal signs preceded the onset of parkinsonian syndrome. In 4 cases, the clinical picture was characterized by the prevalence of motoneuronal signs. Parkinsonism was poorly responsive to L-dopa treatment in all patients. The clinical course did not differ from that expected in patients with only ALS. Our clinical observations and neuropathological reports of nigral neuronal loss in ALS patients suggest a common pathogenic mechanism underlying these disorders. PMID- 12548375 TI - Aggressive angiomyxoma of childhood: two unusual cases developed in the scrotum. AB - Aggressive angiomyxoma (AAM) is a locally invasive soft tissue tumor and is predominantly found in the female pelvis and perineum. The incidence of AAM in adult and adolescent males is low, and is very rare in male children. However, two cases of AAM occurred in the scrotum of 8-year-old and 1-year-old boys. Grossly, both tumors were ill-delineated nodules showing myxoid homogenous and lobulated cut surface. The scrotum of case 2 was replaced by the exuberant mass, making a polypoid appearance. On microscopic examination, both had hypocellular myxoid stroma, spindle and stellate stromal cells, and blood vessels of various calibers. Both lesions typically showed infiltrating borders and penetrated into skin adnexa and Dartos' muscle fibers. AAM should be considered in the differential diagnosis of a scrotal mass found in childhood. PMID- 12548377 TI - Reference values for second trimester fetal and neonatal organ weights and measurements. AB - To establish accurate reference ranges for the entire second trimester, we documented organ weights, body weight, and linear measurements for 597 fetuses and neonates with gestational ages ranging from 12 to 26 wk. We determined the mean and standard deviation for weights and measurements at each week of gestation using the StatView trade mark SE + Graphics statistical program. The analyses revealed a linear correlation between the gestational age and, respectively, the toe-heel length, crown-rump length, and crown-heel length. Body and organ weights increase at varying rates throughout the second trimester. The data correlate well with weights and measurements previously published for the latter half of the second trimester, and extend these reference ranges to encompass the entire second trimester. PMID- 12548379 TI - Prolonged ventilation and intact survival in very low birth weight infants. AB - A requirement for prolonged ventilation (>28 days) has been associated with a poor outcome in infants. We postulated that in the present population of infants who usually receive antenatal steroids and post-natal surfactant, prolonged ventilation in discrete episodes, i.e. discontinuous intermittent positive pressure ventilation (IPPV), would have a better outcome than a requirement for prolonged ventilation continuously from birth (continuous IPPV) and, in addition, that an abnormal ultrasound scan appearance would be a reliable predictor of poor outcome in infants requiring prolonged ventilation. All very low birth weight (VLBW) infants ventilated for at least 28 days (prolonged ventilation) were identified from a prospectively maintained database. At 1 year of age, neurodevelopmental status was assessed and abnormal neurodevelopmental outcome diagnosed if the infant's Griffiths developmental quotient was at least two standard deviations below the mean and/or they had impairment with disability. Of 417 VLBW infants, 41 required prolonged ventilation (30 continuous and 11 discontinuous). In the continuous IPPV group, 18 and one in the discontinuous IPPV group died or had abnormal neurodevelopmental outcome ( P<0.01). All eight infants with major cranial ultrasound abnormalities died or had abnormal outcome ( P<0.01). CONCLUSION: prolonged ventilation can be associated with intact survival, but not in very low birth weight infants with evidence of significant brain injury. PMID- 12548380 TI - Clinical consequences of twin-to-twin transfusion. AB - Twin-to-twin transfusion (TTT) is a complication of monochorionic twins that may result in high mortality and morbidity. To better understand pathophysiology in TTT and the consequences for affected fetuses and neonates, we describe the clinical features of 19 consecutive pregnancies complicated by TTT over 5 years. The diagnosis was made based on the findings of polyhydramnios-oligohydramnios sequence with weight discordance judged by obstetrical sonogram in monochorionic twins. Serial amnioreductions were performed as the sole modality of therapeutic intervention when indicated. The obstetrical diagnosis of TTT was first made at a median gestational age of 26 weeks (range 20-35 weeks). Median age of delivery was 30 weeks (range 26-36 weeks). Thirty-three babies of the 19 pairs of twins were born alive, but only 21 of them lived beyond 28 days of life. Fifteen pairs of twins with TTT had weight discordance greater than 20%, but only one pair showed initial hemoglobin difference greater than 5 g/dl. Newborn infants with TTT were at risk for development of renal insufficiency, periventricular leukomalacia, and necrotizing enterocolitis. Intrauterine fetal demise of one twin and severe anemia (hemoglobin <10 g/dl) at birth were poor prognostic factors. Recent advances in perinatal care improves pregnancy outcome of TTT; however, surviving neonates are still at risk for morbidities arising from hemodynamic aberrations. PMID- 12548381 TI - Serious adverse effects of unconventional therapies for children and adolescents: a systematic review of recent evidence. AB - Unconventional therapies have become popular in paediatric and adolescent populations. It is therefore important to define their risks. The aim of this systematic review was to summarise the recent evidence. Computerised literature searches were carried out in five databases to identify all recent reports of adverse events associated with unconventional therapies in children. The reports were summarised in narrative and tabular form. The results show that numerous case reports and several case series have been published since 1990. Investigations of a more systematic nature are, however, rare. Most of the adverse events were associated with herbal medications. Inadequately regulated herbal medicines may contain toxic plant material, be contaminated with heavy metals, or be adulterated with synthetic drugs. The adverse events included bradycardia, brain damage, cardiogenic shock, diabetic coma, encephalopathy, heart rupture, intravascular haemolysis, liver failure, respiratory failure, toxic hepatitis and death. A high degree of uncertainty regarding a causal relationship between therapy and adverse event was frequently noted. The size of the problem and its importance relative to the well-documented risks of conventional treatments are presently unknown. Several unconventional therapies may constitute a risk to the health of children and adolescents. At present, it is impossible to provide reliable incidence figures. It seems important to be vigilant and investigate this area more systematically. PMID- 12548382 TI - Two siblings with tyrosinaemia type 2. AB - We report on two siblings with tyrosinaemia type 2, a 6-year-old girl and her 3.5 year-old brother with a main complaint of painful palmo-plantar hyperkeratosis. The boy presented also conjunctival injection, photophobia, lacrimation and conjunctivitis. Blood tyrosine levels were increased in both patients, 150.6 and 202.3 micro mol/dl, respectively (reference value: 5.8+/-2.2 micro mol/dl). After 1 month of dietary treatment with low protein, low phenylalanine and tyrosine, tyrosine levels decreased to 37.7 and 65.6 micro mol/dl, respectively; the cutaneous lesions improved in both of them, and conjunctivitis disappeared. CONCLUSION: the association of bilateral ulcero-conjunctivitis with photophobia and palmo-plantar hyperkeratosis since early infancy is the clue to the diagnosis of tyrosinaemia type 2. PMID- 12548383 TI - Effects of anticonvulsants on GLUT1-mediated glucose transport in GLUT1 deficiency syndrome in vitro. AB - Facilitative type-1 glucose transporter (GLUT1) deficiency syndrome is caused by a defect of glucose transport into brain, resulting in an epileptic encephalopathy. Seizures respond effectively to a ketogenic diet, but a subgroup of patients require add-on anticonvulsant therapy or do not tolerate the diet. With the exception of barbiturates, which have been shown to inhibit GLUT1 function, no anticonvulsants have been investigated for possible interactions with GLUT1. Kinetic analyses of (14)C-labeled 3-O-methyl glucose (3OMG) uptake into erythroctes were performed in 11 patients and 30 controls. For in vitro inhibition studies, zero-trans influx of 3OMG (5 mmol/L) into erythrocytes was determined following preincubation with diazepam, carbamazepine, phenytoin, and chloralhydrate. In addition, the effects of ethanol on cell lysis and 3OMG transport into erythrocytes were determined. In patients, mean 3OMG influx was 53% of controls. Ethanol, diazepam, and chloralhydrate significantly inhibited GLUT1 function. Erythrocyte cell lysis was evident at concentrations of 2.5% ethanol. Diazepam, chloralhydrate, and ethanol are inhibitors of GLUT1 function in vitro and might potentiate the effects of GLUT1-mediated glucose transport in patients with GLUT1 deficiency syndrome. In contrast, no inhibitory effects were observed for carbamazepine and phenytoin, indicating that these substances might be preferable for additional seizure control in this disorder. PMID- 12548384 TI - Complete deficiency of mitochondrial trifunctional protein due to a novel mutation within the beta-subunit of the mitochondrial trifunctional protein gene leads to failure of long-chain fatty acid beta-oxidation with fatal outcome. AB - The mitochondrial trifunctional protein (MTP) is a multienzyme complex which catalyses three of the four chain-shortening reactions in the beta-oxidation of long-chain fatty acids. Clinically, failure of long-chain fatty acid beta oxidation leads to hypoketotic hypoglycaemia associated with coma, hepatopathy, skeletal myopathy and cardiomyopathy. We report on consanguineous parents with six children, four of whom had unexpectedly died in Egypt during the neonatal period due to cardiomyopathy of unknown aetiology and respiratory failure. After moving to Germany, two further children died at the age of 4 months and 12 h, respectively, with signs of respiratory and cardiac failure, hydrops fetalis and acidosis. Analysis of acylcarnitine profiles in dried blood spots of the last two children by electrospray tandem mass spectrometry was indicative of a long-chain fatty acid beta-oxidation disorder. Both infants were homozygous for a novel missense mutation (976G-->C) within a highly conserved region of the MTP beta subunit gene. Immunoblot analysis in chorionic villi obtained during the subsequent pregnancy demonstrated absence of MTP. In fibroblasts and liver, activities of all three catalytic units of MTP were markedly decreased, further confirming the diagnosis of MTP deficiency. CONCLUSION: the detected mutation (976G-->C) within the beta-subunit of the mitochondrial trifunctional protein gene destabilises the protein, leading to complete deficiency and a poor prognosis. Immunoblot analysis of mitochondrial trifunctional protein in chorionic villi may be a valuable tool for the prenatal diagnosis of the disorder when the molecular genetic defect is unknown. PMID- 12548385 TI - Diagnosis of juvenile hemochromatosis in an 11-year-old child combining genetic analysis and non-invasive liver iron quantitation. AB - Juvenile or type2 hemochromatosis is a rare autosomal recessive disorder which leads to severe iron overload early in life. As in the classic adult form of the disease iron toxicity causes liver cirrhosis, cardiomyopathy, and endocrine complications, but the onset of the disease is anticipated in the second to third decades of life. Experience of this disease in children is limited. Molecular diagnosis is unfeasible because the type2 hemochromatosis gene is still unknown, although it is known that the disease locus maps to chromosome 1q. Combining linkage analysis with markers encompassing chromosome 1 locus and a non-invasive method for liver iron quantitation we diagnosed juvenile hemochromatosis in a presymptomatic stage in an 11-year-old Italian child. A regular phlebotomy protocol reduced iron overload preventing all the disease complications. CONCLUSION: Juvenile hemochromatosis patients have severe iron overload within the first years of life, strengthening the greater iron absorption that occurs in this as compared to other types of hemochromatosis. Early detection is essential, because treatment in presymptomatic stages prevents organ damage. PMID- 12548386 TI - Interstitial deletion of chromosome 9, int del(9)(9q22.31-q31.2), including the genes causing multiple basal cell nevus syndrome and Robinow/brachydactyly 1 syndrome. AB - We report a child with a de novo interstitial deletion, 46,XY, int del(9)(9q22.31 q31.2). Cytogenetic and molecular analysis defined the boundaries of the lost region, of paternal origin, from D9S1796 to D9S938. The clinical picture included macrocephaly, frontal bossing, bilateral epicanthus, down-slanted palpebral fissures, low-set ears, hypoplastic nostrils, micrognathia, scoliosis, right single palmar crease, small nails, slender fingers, bilaterally flexed 5th finger, delayed bone age, abnormal metacarpophalangeal pattern (MCPP) profile and sole pits. No major malformation was recorded. The deleted region includes, among others, the PTCH and ROR2 genes. Mutations of the former cause the nevoid basal cell carcinoma syndrome (NBCCS) while mutations in the ROR2 gene have been found both in Robinow syndrome and in brachydactyly type 1B (BDB1). As the patient shows some clinical manifestation of both syndromes, we conclude that phenotypic changes related to haploinsufficiency of PTCH and ROR2 are recognisable in our patient even at a young age and in the presence of the more complex phenotype due to the deletion's large size. Thus the efforts to identify the genes included in a deletion are worthy as they may result in better care of the patient as, in this case, monitoring the possible development of tumours associated with NBCCS. PMID- 12548387 TI - Secular trends in body mass index measurements in preschool children from the City of Aachen, Germany. AB - On account of the recent increases in prevalence of childhood obesity in Western countries, the present study tried to verify a secular trend for increasing body mass index (BMI; kg/m(2)) in preschool children in Aachen, Germany. The total sample was based on weight and height data for all 99,500 children of German nationality before enrollment in school in the City of Aachen from 1968-1999. For each year, 10% of the boys and girls respectively, were randomly selected for the analyses. Quantile regression was used to examine the pattern and extent of change in BMI percentiles over this 31-year period. Anthropometric data of a total of 5081 boys and 4863 girls were subjected to quantile regression. While significant increases occurred for any given BMI percentile, the annual increase for both sexes was most prominent in the upper range. No change in body height was observed during the study period. CONCLUSION: preschool children have gained a higher body mass index during the last 30 years. The mechanisms underlying the secular trend towards increasing body mass index seemingly affect children in the upper weight range more than those in the lower range. PMID- 12548388 TI - "Back to the future...": a word from the editor. PMID- 12548389 TI - The ClC-5 chloride channel knock-out mouse - an animal model for Dent's disease. AB - Mutations in the gene CLCN5 encoding the vesicular chloride channel ClC-5 lead to Dent's disease, an X-linked renal disorder. Dent's disease is characterised by proteinuria, hyperphosphaturia and hypercalciuria, which eventually lead to kidney stones and nephrocalcinosis. As it was unclear how mutations in a chloride channel might cause these symptoms, we and others have generated genetic mouse models to elucidate the underlying pathophysiological mechanisms. We review results obtained from these three mouse models and present new data on endosomal acidification and vitamin D metabolism in ClC-5 knock-out (KO) mice. ClC-5 is expressed in apical endosomes of proximal tubular cells where it co-localizes with endocytosed proteins and the proton ATPase. ClC-5 may provide an electric shunt for the efficient operation of the electrogenic H(+)-ATPase. We confirmed this hypothesis by showing that endosomes from CLCN5 KO mice are acidified at a significantly lower rate than wild-type endosomes. This probably results in the drastic impairment of endocytosis observed in ClC-5 KO mice. Parathyroid hormone (PTH) is filtered into the lumen of the nephron, where it is endocytosed and degraded by proximal tubular cells. The defective endocytosis in ClC-5 KO mice entails an increased luminal concentration of PTH, subsequent stimulation of apical PTH receptors which causes an increased endocytosis of the phosphate transporter NaPi and phosphaturia. We now show that it also results in up regulation of proximal tubular alpha-hydroxylase that generates the active form of vitamin D from its precursor. We discuss how the primary defect in endocytosis leads via secondary changes in calciotropic hormones to the tertiary symptoms hyperphosphaturia, hypercalciuria and kidney stones. PMID- 12548390 TI - Inactivation of sodium-transporting proteins in the kidney. AB - The kidney plays a dominant role in maintaining sodium homeostasis. The control of a nearly constant electrolyte composition and osmotic pressure in the extracellular fluids is achieved by well-regulated vectorial salt and water transport processes. Derangement in function of Na(+) transporting proteins is likely to be responsible for a number of clinical disorders of fluid and electrolyte homeostasis. The identification of the genes implicated in sodium reabsorption in the kidney not only allows a detailed analysis of regulation and function of these proteins in vitro but also the generation of genetically engineered mice that constitute valuable mouse models for human diseases. Our review will focus on recent strategies for generating nephron segment-specific knock-outs for the main apical renal Na(+) transporters and channels. PMID- 12548391 TI - Tubuloglomerular feedback in the kidney: insights from gene-targeted mice. AB - The term tubuloglomerular feedback (TGF) refers to a series of events whereby changes in the concentration of Na(+), Cl(-) and K(+) in the distal tubular fluid are sensed by the macula densa and elicit reciprocal changes in nephron filtration rate (SNGFR). Although micropuncture experiments in the rat and studies in the isolated perfused rabbit juxtaglomerular apparatus (JGA) have yielded significant insights into TGF, many aspects, including the signal transduction and modulation in JGA and the role of TGF in physiology and pathophysiology, are still incompletely understood. This is due in part to the lack of selective drugs for inhibiting or activating a given molecule of interest and to the limited accessibility of target structures or molecules. Adaptation of single-nephron methods to gene-targeted mice is a promising approach for gaining further insights into TGF. The gist of the presently available studies in gene targeted mice that are reviewed here support the concept that (i) adenosine mediates TGF by activation of adenosine A(1) receptors; (ii) that this response is modulated inversely by nitric oxide synthase 1 activity in macula densa and activation of angiotensin II AT(1A) receptors; and (iii) that the TGF-dependent reduction in SNGFR prevents excessive renal fluid and salt loss during primary inhibition of reabsorption upstream from the macula densa. PMID- 12548392 TI - Pathophysiological role of the mineralocorticoid receptor in heart: analysis of conditional transgenic models. PMID- 12548393 TI - A gene knockout approach in mice to identify glucose sensors controlling glucose homeostasis. AB - A key aspect of glucose homeostasis is the constant monitoring of blood glucose concentrations by specific glucose sensing units. These sensors, via stimulation of hormone secretion and activation of the autonomic nervous system (ANS), regulate tissue glucose uptake, utilization or production. The best described glucose detection system is that of the pancreatic beta-cells which controls insulin secretion. Secretion of other hormones, in particular glucagon, and activation of the ANS, are regulated by glucose through sensing mechanisms which are much less well characterized. Here I review some of the studies we have performed over the recent years on a mouse model of impaired glucose sensing generated by inactivation of the gene for the glucose transporter GLUT2. This transporter catalyzes glucose uptake by pancreatic beta-cells, the first step in the signaling cascade leading to glucose-stimulated insulin secretion. Inactivation of its gene leads to a loss of glucose sensing and impaired insulin secretion. Transgenic reexpression of the transporter in GLUT2/beta-cells restores their normal secretory function and rescues the mice from early death. As GLUT2 is also expressed in other tissues, these mice were then studied for the presence of other physiological defects due to absence of this transporter. These studies led to the identification of extra-pancreatic, GLUT2-dependent, glucose sensors controlling glucagon secretion and glucose utilization by peripheral tissues, in part through a control of the autonomic nervous system. PMID- 12548394 TI - Current successes and limitations of using genetic modification for blood pressure research. PMID- 12548395 TI - Regulation of airway surface liquid volume by human airway epithelia. AB - Mucus clearance on airway surfaces is a primary form of pulmonary defense. The efficiency of mucus clearance in large part depends on the volume of the airway surface liquid components, including both the periciliary liquid (PCL) layer and the mucus layer. Studies with in vitro model systems suggest that the mucus layer acts as a passive reservoir to redistribute water to and from, as needed, the PCL layer. In contrast, the overall volume of airway surface liquid is determined by active transepithelial salt transport. Data from in vitro systems suggest that airway epithelia have the capacity to both absorb and secrete liquid in response to the volume requirements on the apical surface. At present, the nature of the signals that transmit information about airway surface liquid volume to epithelia and their sensors are unknown. However, progress in elucidation of this system is important, because it appears that these systems are deranged in the genetic disease cystic fibrosis, which is characterized by airway surface liquid volume depletion, mucus stasis, and chronic infection. Thus, insights into these systems may offer novel therapeutic opportunities to correct this physiologic dysfunction of airway epithelia. PMID- 12548396 TI - Functional interaction of CFTR and ENaC in sweat glands. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) plays a significant role in transepithelial salt absorption as well as secretion by a number of epithelial tissues including sweat glands, airways and intestine. Early studies suggested that in absorption significant cross talk occurs between CFTR Cl(-) channels and epithelial Na(+) channels (ENaC). Studies based primarily on cultured cells of the airways and on ex vivo expression systems suggested that activating CFTR inhibits ENaC channels so that activation of CFTR and deactivation of ENaC seem reciprocal. Lack of CFTR Cl(-) conductance (g(CFTR)) in the plasma membranes was seen to enhance ENaC conductance (g(ENaC)) and Na(+) absorption from the airway surface liquid causing airway pathology in cystic fibrosis (CF). To determine if these events hold true for a purely absorptive epithelium, we investigated the role of CFTR in regulating g(ENaC) in native human sweat gland ducts. After permeabilizing the basilateral membrane of the duct with alpha-toxin, the relative activities of ENaC and CFTR in the apical membrane were characterized by correlating the effect of activating CFTR with ENaC function. We found that in contrast to reciprocal activities, activating g(CFTR) by either cAMP, cGMP or the G-proteins plus 5 mM ATP was accompanied by a concomitant activation, not inhibition, of g(ENaC). The activation of g(ENaC) appeared to be critically dependent on CFTR Cl(-) channel function because removal of Cl(-) from the medium, blockage of CFTR with inhibitor DIDS or the absence of CFTR in the DeltaF508 CF ducts prevented activation of g(ENaC) by cAMP, GMP or G-proteins. Most significantly, g(ENaC) was dramatically reduced, not increased, in CF as compared to non-CF sweat ducts. These results showed that lack of CFTR in the plasma membranes is not characteristically coupled to elevated ENaC activity or to increased Na(+) absorption in CF epithelial cells. Not only are CFTR and ENaC activated together in duct salt absorption, but ENaC activation depends on functioning CFTR. NaCl is poorly absorbed in the CF duct because CFTR activity appears to impose a loss of ENaC activity as well. PMID- 12548397 TI - ENaC is inhibited by an increase in the intracellular Cl(-) concentration mediated through activation of Cl(-) channels. AB - Activation of the CFTR Cl(-) channel inhibits epithelial Na(+) absorption, according to studies on native epithelia derived from airways, colon and kidney, and can also be demonstrated in overexpressing cells. However, Na(+) absorption is not inhibited by CFTR in the native sweat duct epithelium. The mechanism for the inhibition of epithelial sodium channels (ENaC) has been examined in most detail in Xenopus oocytes coexpressing CFTR and ENaC. It was shown that ENaC is inhibited during stimulation of CFTR in Xenopus oocytes, independent of the experimental setup and the magnitude of the whole-cell current. However, a minimal Cl(-) conductance is required for inhibition of ENaC, and inhibition is augmented at higher CFTR-to-ENaC currents ratios. Low-CFTR-to-ENaC conductance ratios may be the reason for the absence of ENaC inhibition, as described recently. Similar to CFTR, ClC-0 Cl(-) currents also inhibit ENaC, as well as high extracellular Na(+) and Cl(-) in partially permeabilized oocytes. Thus, inhibition of ENaC is not specific to CFTR and could be mediated by Cl(-) flow and/or changes in the intracellular Cl(-) concentration. These results are reminiscent of the Cl(-) feedback regulation observed in mouse mandibular duct cells. Current results obtained with ENaC mutants examined in Xenopus oocytes suggest a charge interaction of Cl(-) ions with the epithelial sodium channel. PMID- 12548398 TI - cAMP-dependent activation of CFTR inhibits the epithelial sodium channel (ENaC) without affecting its surface expression. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is thought to modulate epithelial sodium channel (ENaC) function in various preparations. However, the molecular nature and (patho-)physiological significance of the CFTR/ENaC interaction is still unclear and may vary in different tissues. Co expression experiments in Xenopus laevis oocytes are a popular approach to investigate a possible functional interaction of CFTR and ENaC but have revealed controversial results. We could confirm previous reports that in oocytes co expressing ENaC and CFTR the amiloride-sensitive current was reduced during cAMP mediated stimulation of CFTR. In contrast, co-expression of CFTR per se had no effect on baseline ENaC currents. ENaC with Liddle's syndrome mutation is also inhibited during activation of CFTR, suggesting that the C-terminus of the ENaC beta-subunit is not important for this functional interrelation. Single-channel patch-clamp recordings demonstrated that co-expression of CFTR does not alter the single-channel conductance of ENaC. Using a chemiluminescence assay we demonstrated that the inhibition of ENaC during cAMP-dependent activation of CFTR was not associated with a decrease in ENaC surface expression. We conclude that the inhibitory effect of cAMP-activated CFTR on ENaC is due to a decrease in channel open probability. PMID- 12548399 TI - ENaC-CFTR interactions: the role of electrical coupling of ion fluxes explored in an epithelial cell model. AB - CFTR and the epithelial Na channel (ENaC) are two membrane proteins expressed in the apical membrane of several epithelia. They have been shown to influence each other's function but the mechanisms of this interaction are not fully understood. We have explored the role of electrical coupling between the ion fluxes carried by these two electrogenic transporters as one among other possible mechanisms through which these two proteins could interact. For this purpose, we have built a mathematical model of an epithelial cell monolayer comprising variable apical Cl(-) and Na(+) conductances, a basolateral Na,K-pump, basolateral Cl(-) and K(+) conductances and a paracellular "shunt" conductance. The model allows us to simulate ion transport under current- or voltage-clamp conditions and to monitor individual ionic membrane currents, membrane potentials and intracellular ion concentrations. Using a set of transport parameters extrapolated from published work, we can show, first, that the model reproduces the expected effects of conductance inhibition on transepithelial transport, and transepithelial and single membrane potentials or conductances. Second, concerning the influence of CFTR on ENaC, the simulations show that: (1) under current-clamp conditions, the amplitude of the apical membrane Cl(-) conductance has a strong influence on the transepithelial depolarization induced by inhibition of the apical Na(+) conductance, (2) under voltage-clamp conditions, the decrease of short-circuit current induced by inhibition of the apical Na(+) conductance is also significantly decreased by the presence of a large apical membrane Cl(-) conductance. Our results show that when ENaC and CFTR are expressed in the same apical membrane, the electrical coupling of these two ion channels can explain at least part of the effect of CFTR activation on the Na(+) current flowing through ENaC. PMID- 12548400 TI - Colocalisation of the protein tyrosine phosphatases PTP-SL and PTPBR7 with beta4 adaptin in neuronal cells. AB - The mouse gene Ptprr encodes the neuronal protein tyrosine phosphatases PTP-SL and PTPBR7. These proteins differ in their N-terminal domains, with PTP-SL being a cytosolic, membrane-associated phosphatase and PTPBR7 a type I transmembrane protein. In this study, we further explored the nature of the PTP-SL-associated vesicles in neuronal cells using a panel of organelle markers and noted a comparable subcellular distribution for PTP-SL and the beta4-adaptin subunit of the AP4 complex. PTP-SL, PTPBR7 and beta4-adaptin are localised at the Golgi apparatus and at vesicles throughout the cytoplasm. Immunohistochemical analysis demonstrated that PTP-SL, PTPBR7 and beta4-adaptin are all endogenously expressed in brain. Interestingly, coexpression of PTP-SL and beta4-adaptin leads to an altered subcellular localisation for PTP-SL. Instead of the Golgi and vesicle type staining pattern, still observable for beta4-adaptin, PTP-SL is now distributed throughout the cytoplasm. Although beta4-adaptin was found to interact with the phosphatase domain of PTP-SL and PTPBR7 in the yeast two-hybrid system, it failed to do so in transfected neuronal cells. Our data suggest that the tyrosine phosphatases PTP-SL and PTPBR7 may be involved in the formation and transport of AP4-coated vesicles or in the dephosphorylation of their transmembrane cargo molecules at or near the Golgi apparatus. PMID- 12548401 TI - Quantitative measurement of cell migration using time-lapse videomicroscopy and non-linear system analysis. AB - Epithelial cells of the mammary gland possess the inherent capacity to form epithelial monolayers in vitro. This requires coordination of cell migration, cell-cell contact formation, and cell proliferation. Using time-lapse phase contrast videomicroscopy we have observed mammary gland epithelial cells over different time scales. We show the generation of a complete polarized epithelial monolayer in real-time, starting from a few cells. We subsequently concentrated on the early stages of this process by tracking epithelial cells during phases of polarized migration. We performed migration analysis using fractal measures. With this technology the structure of seemingly random processes not accessible to the usual methods of linear analysis can be measured. As a control and proof of principle approach we applied infection of cells with an adenoviral vector, which is used as a gene targeting vector for many applications. Infection markedly influenced the patterns of migratory behavior. We, therefore, believe that time lapse videomicroscopy in combination with fractal analysis can contribute to differential characterization of distinct cellular migration patterns. This will be useful in situations of long-term alterations in cell culture systems. PMID- 12548402 TI - Expression of connexin 43 and ZO-1 in differentiating ameloblasts and odontoblasts from rat molar tooth germs. AB - We studied the distribution of connexin (Cx) 43 and ZO-1 by confocal laser scanning microscopy at early stages of dentinogenesis and amelogenesis. Labeling for Cx43 was observed at early stages of differentiation in both the epithelial cells and differentiating odontoblasts. Immunolabeling was detected at the distal and medial regions of undifferentiated ameloblasts and between cells from stratum intermedium and stellate reticulum. Differentiating odontoblasts exhibited immunoreaction for this antibody at their distal end. Immunoreactivity for ZO-1 was observed at regions that correspond to the proximal and distal junctional complexes of differentiating ameloblasts. Staining for ZO-1 was observed at apical regions of odontoblasts with a punctate appearance. In more advanced stages, expression of Cx43 was more evident on ameloblasts, especially at the junctional complexes. Punctate immunolabeling for Cx43 was observed at the lateral sides of differentiating ameloblasts and between the other cells of the enamel organ. Immunoreaction for ZO-1 in ameloblasts was more evident than at the previous stage. It was also observed at the distal end of differentiated odontoblasts. The present study showed that differentiating ameloblasts and odontoblasts express Cx43 and ZO-1 as early as the start of the differentiation process. In addition, the expression of these junctional proteins increases as differentiation of cells continues. PMID- 12548403 TI - An immunohistochemical study on hard tissue formation in a subcutaneously transplanted rat molar. AB - While dental pulp undergoes calcification following tooth replantation or transplantation, we actually know little about these mechanisms. We therefore conducted histological and immunohistochemical evaluations of mineralized tissue that formed in the pulp of rat maxillary molar transplanted into abdominal subcutaneous tissue. One, 2, 3, and 4 weeks post-transplantation, the teeth were investigated immunohistochemically using antibodies to osteocalcin (OCN), osteopontin (OPN), bone sialoprotein (BSP), dentin sialoprotein (DSP), and tissue non-specific alkaline phosphatase (TNAP). In the 1st week after transplantation, cell-rich hard tissue was formed at the root apex. At 2 weeks, formations of hard tissue, with few cells in the root canals and bone-like tissue in the coronal pulp chamber, were noted. After 3 and 4 weeks, the amounts of these hard tissues were increased. The immunolocalization of OCN, OPN, and BSP was seen strongly in coronal and apical hard tissues, but weakly in the root hard tissue. Conversely, DSP localized in the root hard tissue, but not in other newly formed hard tissues. At 1 week, TNAP localized along the periphery of the apical hard tissue and the lower surfaces of root predentin. These results demonstrate that the newly formed hard tissues in the pulp cavity of subcutaneously transplanted molars could be classified into three types, suggesting that these might be formed by type-specific cells. PMID- 12548404 TI - Expression of the peptide transporter hPepT1 in human colon: a potential route for colonic protein nitrogen and drug absorption. AB - Substrates of the proton-coupled peptide transporter, hPepT1, include dietary di- and tripeptides plus therapeutically important drugs such as the beta-lactam antibiotics and angiotensin-converting enzyme inhibitors. Expression and function of hPepT1 in the small bowel is well established. We have compared levels of hPepT1 mRNA expression in regions of human gut by RT-PCR methods and examined the expression of hPepT1 in normal human colon using an anti-hPepT1 antipeptide antibody. hPepT1 mRNA was expressed in the large intestine, although at lower levels than in the small intestine. Quantitatively, expression in ileum was 4.6 fold greater than in sigmoid colon. Immunoreactive hPepT1 was detected in human colon at lower levels than in ileum. The pattern of expression differed between the two tissues: whilst expression in the ileum was localised to the apical enterocyte membrane along the length of the crypt-villus axis, expression in the colonocyte was detected at the apical membrane towards the luminal surface but predominantly at the basal membrane towards the base of the crypt. We conclude that distal regions of the bowel express hPepT1, which may provide a mechanism for colonic protein-nitrogen absorption and for absorption of therapeutically important peptidomimetic drugs. PMID- 12548405 TI - Expression of IFI 16 in epithelial cells and lymphoid tissues. AB - IFI 16 is a member of the HIN-200 protein family named for their haemopoietic expression, interferon-inducibility and nuclear localisation. These proteins have been characterised as transcriptional regulators that modulate the cell cycle. IFI 16 is expressed in some haemopoietic lineages including CD34+ progenitor cells, mature lymphocytes and monocytes, but is absent from granulocytes, erythrocytes and megakaryocytes. We present a wider study of IFI 16 expression in normal human tissues using a monoclonal antibody specifically recognising the C terminus of IFI 16. As expected, IFI 16 was detected in the nuclei of lymphocytes in the spleen, thymus, lymph node and palatine tonsil, but was also found in epithelial cells in these tissues. Interestingly, IFI 16 protein was also expressed in non-lymphoid tissues including trachea, gastrointestinal tract, skin and testis, but was absent from others including heart and brain. In each tissue, IFI 16 was predominantly expressed in surface epithelial cells and staining was strongest in basal epithelial layers. Therefore, IFI 16 expression is not restricted to cells of the immune system, but is also expressed in epithelial cells. In contrast to the perceived role of HIN-200 proteins as suppressors of cell growth, maximal expression of IFI 16 was in cells with high proliferative potential. PMID- 12548406 TI - Upregulation of TrkA neurotrophin receptor expression in the thymic subcapsular, paraseptal, perivascular, and cortical epithelial cells during thymus regeneration. AB - Neuroimmune networks in the thymic microenvironment are thought to be involved in the regulation of T cell development. Here, we report upon an examination of the expression of the TrkA neurotrophin receptor, the high affinity receptor for nerve growth factor, during regeneration following acute involution induced by cyclophosphamide in the rat thymus. Light and electron microscopic immunocytochemistry demonstrated enhanced expression of the TrkA receptor in the subcapsular, paraseptal, perivascular, and cortical epithelial cells during thymus regeneration. In addition, various morphological alterations, suggestive of a hyperfunctional and dynamic state, of the subcapsular, paraseptal, and perivascular epithelial cells were also observed. The presence of TrkA protein in extracts from the control and regenerating rat thymus was confirmed by western blot. Furthermore, RT-PCR analysis supported these results by demonstrating that thymic extracts contain TrkA mRNA at higher levels during thymus regeneration. Thus, our results suggest that the TrkA receptor located on the thymic subcapsular, paraseptal, perivascular, and cortical epithelial cells could play a role in the development of new T cells to replace T cells damaged during thymus regeneration. PMID- 12548407 TI - Occurrence and localization of uroguanylin in the aging human prostate. AB - Uroguanylin, a peptide hormone highly expressed in the gastrointestinal tract, is implicated in the regulation of epithelial salt and water transport processes. Since little is known about a possible role of uroguanylin in the reproductive system, we investigated for the first time the occurrence of this peptide in the human prostate using specimens of benign prostatic hyperplasia. Northern blot analyses detected a single uroguanylin transcript of approximately 600 bp in prostate RNA. The uroguanylin expression was further investigated by reverse transcriptase polymerase chain reaction of prostate RNA with uroguanylin-specific primers. Sequencing of the fragments obtained indicated the presence of a uroguanylin molecule with a sequence identical to its intestinal counterpart. Furthermore, in situ hybridization and immunohistochemistry revealed that uroguanylin mRNA and peptide are confined to epithelial cells of the prostate glands. Comparison with the distribution pattern of immunoreactivity for prostate specific antigen (PSA) showed a high degree of colocalization of uroguanylin- and PSA-immunoreactive cells. In addition, by western blotting techniques we detected the presence of high molecular weight uroguanylin-immunoreactive material in prostatic fluid. In conclusion, our study indicates that the human prostate glands synthesize and secrete (pro-)uroguanylin. We hypothesize that this hormone may play a novel role in the male reproductive tract. PMID- 12548409 TI - Chronic pancreatitis: etiology, pathogenesis, diagnosis, and treatment. AB - BACKGROUND AND AIMS: This review discusses current concepts regarding the etiology, pathogenesis, diagnosis, and treatment of chronic pancreatitis. RESULTS: Treatment of patients suffering from complications of chronic pancreatitis remains a major challenge. Continuous alcohol consumption, a disease with enormous personal and social impact, is still the leading factor in the development of chronic pancreatitis. The most distressing symptom for the patient is pain, which in many instances has already led to a considerable analgesic abuse before a specialist is even consulted. The development of an enlargement of the pancreatic head, the "pacemaker of the disease," parallels pain and potential organ complications such as common bile duct stenosis or portal hypertension. What triggers pancreatic head enlargement is still not known; in particular it is unclear what triggers growth factors to step into action. Pain is most likely due to a combination of hypertension in the organ and parenchymal alterations. Current therapeutic concepts therefore aim at alleviating pain and at management of organ complications while preserving functional tissue to best prevent exocrine and endocrine dysfunction. The ideal procedure thus seems today to be the "customized" pancreatic head resection with additional optional drainage of the pancreatic body and tail. PMID- 12548408 TI - Different apoptotic responses and patterns in adhering and floating neoplastic cell cultures: effects of microtubule antagonists. AB - The relationship between apoptotic progression and cell cycle perturbation induced by microtubule-destabilising (vinblastine, Colcemid) and -stabilising (taxol) drugs was studied in two mesenchyme-derived neoplastic cell lines, growing as suspension (Jurkat) and monolayer (SGS/3A) culture, by morphocytochemical and biochemical approaches. The same kind of drug induced different effects on the cell kinetics (proliferation, polyploidisation, death) of the two cell lines. In floating cells, the drugs appeared more effective during the S phase, while in adherent cells they were more effective during the G2/M phase. Moreover two distinct neoplasia-associated apoptotic phenotypes emerged: the first pattern was the typical one and was found in cells with a low transition through the S/G2 phase (Jurkat), and the second one was mainly characterised by a cell death derived from micronucleated and mitotic cells, as a consequence of a low transition through the M/G1 phase (SGS/3A). Our data show that the machinery required for the trigger and progression of apoptosis is present in every cell cycle phase, also in conditions of karyological alterations (aneugenic micronucleations). On the other hand, a different sensitivity of the two microtubular components (interphasic network and mitotic spindle) appears to be related to the anchorage-dependence or -independence during the cell growth disturbances after exposure to antimicrotubular drugs. PMID- 12548410 TI - Randomized clinical trial comparing simple drainage of anorectal abscess with and without fistula track treatment. AB - BACKGROUND AND AIMS: Anal abscess is a frequent acute proctological disorder and whether the underlying fistula should be treated at the same time when the abscess is drained remains controversial. We examined indications for drainage alone versus drainage plus fistulotomy in terms of recurrence and continence. PATIENTS AND METHODS: We carried out a randomized prospective study of 200 consecutive patients with anal abscess. One group received drainage alone, while in the other group drainage plus fistulotomy was performed when a subcutaneous mucosa, low transsphincteral, or intersphincteral fistula was found. Delayed progressive fistulotomy with suture threads was performed in cases of high transsphincteric or suprasphincteric fistula. RESULTS: The internal opening of the fistula track was found in 83% of the patients. The recurrence rate was related to the surgical technique employed: 29% in the group with drainage alone and 5% in the group for which treatment of the fistula track was attempted. The incontinence rate was also related to the surgical option. In those receiving drainage and treatment of the fistula track incontinence was restricted mostly to patients with delayed fistulotomy (36.7%), compared to 2.8% of patients when simple fistulotomy was performed. There was no incontinence in the drainage alone group. CONCLUSION: Drainage of anal abscess with fistulotomy can be safely performed in cases of subcutaneous, intersphincteral, or low transsphincteral fistulae with a minimal recurrence rate. However, drainage alone and posterior treatment of the fistula track is recommended for high transsphincteral or suprasphincteral fistulae. PMID- 12548411 TI - Change in anal continence after surgery for intersphincteral anal fistula: a functional and manometric study. AB - BACKGROUND AND AIMS: Dividing or "laying open" of the tract for intersphincteral type anal fistula is simple and effective and entails low risk of complication, but little is known of the functional and manometric results. This study assessed the clinical and manometric effect of this surgery on anal sphincter function. PATIENTS AND METHODS: The study examined 45 adults undergoing surgical treatment for intersphincteral fistula. We administered the questionnaire for continence score and performed anorectal manometry before the operation and at least 6 months after the operation. The operative method was laying open of the fistula tract and trimming the redundant anoderm for adequate drainage of the wound in all cases. RESULTS: There was a significant decrease in maximal resting anal pressure and in resting pressure throughout the distal 2 cm of the anal canal after operation. The maximal contractile pressure after operation was similar to that before operation. Continence control was significantly poorer in women and patients who had lower preoperative resting pressure. Multivariate analysis showed lower preoperative resting pressure to be the only independent factor for impaired continence control after fistula surgery. CONCLUSION: Although laying open of the fistula tract is a simple and effective therapy for intersphincteral type anal fistula, it should be more conservative for patients with low resting anal pressure. PMID- 12548412 TI - How consistent is the anal transitional zone in the double-stapled ileoanal reservoir? AB - BACKGROUND AND AIMS: The double-stapled ileoanal reservoir (DSIAR) has become a preferred method for ileal pouch anal anastomosis in restorative proctocolectomy. This investigation assessed the relative ratio of epithelial tissue types within the anal transition zone after a DSIAR and reviewed functional physiological differences. PATIENTS AND METHODS: All 138 patients who underwent restorative proctocolectomy with a DSIAR for mucosal ulcerative colitis were stratified into two groups according to histological evidence of epithelium types in the distal excised rectal donut. In group I a squamous or a squamous mixed with columnar epithelium was present (n=40) whereas in group II only columnar epithelium was seen (n=98). Anal physiological parameters were measured by anal manometry preoperatively, prior to ileostomy closure, and 1 year after surgery. RESULTS: None of the preoperative resting and squeeze pressure parameters showed a significant difference between the two groups. Postoperative mean and maximal resting pressures were significantly decreased in both groups. Postoperative mean and maximum squeeze pressures were decreased in group I. Postoperative mean length of the high-pressure zone tended to be decreased in both groups. The decrease in rectoanal inhibitory reflex was significant in both groups. Postoperative functional parameters measured as maximal tolerable volume and compliance were significantly improved from preoperative values in both groups. CONCLUSION: The tissue type in the anal transitional zone after DSIAR has a wide variability at a similar level (height) of the anastomosis. However, these different epithelial types were not associated with substantial physiological functional differences. Thus, if technically feasible, it is desirable for DSIAR to construct the anastomosis as close to the dentate line as possible. PMID- 12548413 TI - Computed tomography enteroclysis in comparison with ileoscopy in patients with Crohn's disease. AB - BACKGROUND AND AIMS: Enteroclysis and computed tomography (CT) have been recently combined in to assess small bowel alterations. We compared the accuracy of CT enteroclysis to that of endoscopy in detecting bowel wall alterations of the terminal or neoterminal ileum in Crohn's disease (CD) patients and assessed whether postcontrast wall density is related to clinical activity of CD. PATIENTS AND METHODS: A total of 39 patients referred for either established or suspected CD were enrolled. Diagnosis used ileocolonoscopy with histology; clinical activity was measured by CDAI. Contrast-enhanced spiral CT of the abdomen was performed after distension of the small bowel with an enema of methylcellulose. Retrograde ileocolonoscopy diagnosed 30 patients with CD of the ileum, while 9 patients served as controls. RESULTS: CT enteroclysis detected CD in 26 patients (86.7%) and in none of the control group. Three of four patients with false negative findings on CT enteroclysis had postsurgical CD recurrence. The overall sensitivity and specificity of CT enteroclysis for ileal CD detection were 86.7% and 100%, respectively (PPV=100%; NPV=69.2%), and 94.1% and 100% (PPV=100%; NPV=90%), excluding those patients with postsurgical recurrence. The postcontrast wall density was significantly higher in CD patients than in the controls and was significantly correlated with the severity of CD. CONCLUSION: CT enteroclysis proved highly accurate in detecting terminal ileal CD involvement, particularly in patients without previous surgery, and to allow assessment of the degree of intramural and clinical activity. PMID- 12548414 TI - Should routine spiral computed tomography be performed in adult patients with acute abdominal pain of the right lower quadrant? AB - BACKGROUND AND AIMS: To determine whether spiral computed tomography (SCT) is helpful in the management of adult patients with acute pain of the right lower quadrant. PATIENTS AND METHODS: The prospective study included 204 patients with acute pain of the right lower quadrant during two periods: 104 before and 100 after the introduction of SCT. A questionnaire was completed for each patient before and after standard clinical and laboratory assessment and SCT, indicating the suspected diagnosis and a provisional therapeutic option. RESULTS: In the pre SCT period the negative laparotomy rate was 29% vs. only 13% in the SCT period. During the SCT period 10 of the 24 nonoperated patients were discharged on the same day, and 14 were hospitalized 1.6+/-1 days, and only one was readmitted for operation on day 7. The accuracy of SCT assisted by results of clinical and laboratory findings (true positives + true negatives/total of patients) was 86% (vs. 73% for standard clinical and laboratory assessment): 93% in men (vs. 76% for standard clinical and laboratory assessment) but only 63% in women (vs. 60% for standard clinical and laboratory assessment). SCT changed the therapeutic decision guided by standard clinical and laboratory assessment in 18 cases, accurately in 16 cases (90%). CONCLUSION: SCT is helpful in the management of adult patients with acute abdominal pain of the right lower quadrant. In our study it reduced negative laparotomy rate from 29% (guided by standard clinicobiological evaluation) to only 13%. PMID- 12548415 TI - Endoscopic snare resection of large colonic polyps: how far can we go? AB - BACKGROUND AND AIMS: Colonoscopic polypectomy is preventing colorectal cancer. Videoendoscopy and new perendoscopic hemostasis techniques make endoscopic polypectomy of large colonic polyps an alternative to the surgical approach. This study examined whether complete snare resection of giant colonic polyps is feasible and safe and for determining how often surgery is necessary due to invasive cancer detected histologically after polypectomy. PATIENTS AND METHODS: The study included 59 consecutive patients with 68 colonic polyps larger 30 mm in diameter. Snare polypectomy was performed after an endoscopic ultrasound with a miniprobe found no sign of invasive, or, depending on the appearance of the polyp, a bleeding prophylaxis had been carried out. Acute procedural or delayed bleeding was treated endoscopically. RESULTS: Of the 68 polyps 26, mostly pedunculated were resected en bloc (38%) and histologically ensured as completely resected; 42 polyps had to be resected by piecemeal technique (62%). Piecemeal resection was performed significantly more often in sessile polyps (38/41, 93%) than in pedunculated polyps (4/27, 15%, P<0.01). Follow-up colonoscopy after 3 months showed remaining adenomatous tissue of piecemeal-resected polyps in 12 cases (28%), which were 12 resected sessile polyps (29%) and no case of resected pedunculated polyp. To achieve complete resection of sessile polyps a second procedure was necessary significantly more often than for resection of pedunculated polyps (12 cases in sessile polyps, 18% vs. no case in pedunculated polyps). Remaining adenomatous tissue was removed in all 12 cases during the first follow-up colonoscopy after 3 months, confirmed by a biopsy 6 months after the initial procedure. Overall coexisting malignancy was found in only 7 polyps (12%). Due to high-risk factors only one of them underwent secondary surgical procedure. CONCLUSION: The present study shows that endoscopic snare resection of giant colonic polyps is a safe procedure, and that secondary operative measures for managing coexisting malignancy are rarely necessary. PMID- 12548416 TI - Effect of electromagnetic fields and early postoperative 5-fluorouracil on the healing of colonic anastomoses. AB - BACKGROUND AND AIMS: Studies have indicated a deleterious effect of perioperative 5-fluorouracil (5-FU) administration on the healing of intestinal anastomoses. This study examined the effect of early postoperative 5-FU on the healing of colonic anastomoses and investigated the effect of electromagnetic fields (EMF) on colonic anastomotic repair under normal physiological conditions and in the presence of 5-FU therapy in a rat model. MATERIALS AND METHODS: Forty male Wistar rats were randomly assigned into four groups and underwent a standardized left colonic resection and anastomoses. The animals then served as control or received intraperitoneal 5-FU (20 mg/kg per day, 5 days), EMF stimulation (10.76 mT, 50 Hz; 2-h on/10-h off cycles, 7 days) or both, starting on the day of surgery. After 7 days anastomotic healing was assessed by measurement of hydroxyproline content and breaking strength. RESULTS: Hydroxyproline content increased in EMF exposed group (1.53+/-0.11 to 1.92+/-0.11 microg/mg) and in EMF + 5-FU group (1.53+/-0.11 to 1.89+/-0.12 microg/mg). Breaking strength also increased in the EMF group (0.23+/-0.02 to 0.27+/-0.01 MPa) and in the EMF + 5-FU group (0.23+/ 0.02 to 0.28+/-0.01 MPa. No differences were found in hydroxyproline content or breaking strength between the 5-FU group and controls. CONCLUSION: Early postoperative 5-FU administration did not impair the healing of colonic anastomoses in rats. Additionally, EMF stimulation provided a significant gain in colonic anastomotic strength, in rat intestines in control animals and in animals exposed to 5-FU. PMID- 12548417 TI - Plasma levels of soluble CD40 ligand are elevated in inflammatory bowel diseases. AB - BACKGROUND AND AIMS: CD40/CD40 ligand (CD40L) interaction is important for induction of T cell dependent antibody production and cell-mediated immune responses. Overexpression of CD40/CD40L in the intestinal mucosa is likely to be involved in the pathogenesis of inflammatory bowel disease (IBD). A soluble form of CD40L (sCD40L) exists in the circulation. This study investigated whether plasma levels of sCD40L are higher in patients with IBD than in healthy controls. PATIENTS AND METHODS: Plasma levels of sCD40L were measured in 89 patients with Crohn's disease (CD), 56 patients with ulcerative colitis (UC), 17 patients with infectious diarrhea, and 42 healthy controls, using a specific enzyme-linked immunosorbent assay. RESULTS: In CD patients plasma levels of sCD40L were significantly higher than in healthy controls. Patients with UC and infectious diarrhea had higher sCD40L levels than healthy controls, but the differences were not significant. CD patients with fistulas and/or abscesses (n=38) had significantly higher levels of sCD40L than patients with uncomplicated CD (n=51). Only in patients with uncomplicated CD plasma levels of sCD40L correlated significantly with C-reactive protein and alpha(1)-glycoprotein. In UC patients there was a significant correlation of sCD40L with C-reactive protein. However, there was no significant correlation between plasma sCD40L levels and Crohn's disease activity index or Rachmilewitz score. CONCLUSION: Elevated plasma levels of sCD40L in CD patients supposedly reflect activation of functional CD40L in the intestine and might be a marker of intestinal inflammation. PMID- 12548418 TI - Elevated plasma gastrin, CEA, and CA 19-9 levels decrease after colorectal cancer resection. AB - BACKGROUND AND AIMS: Gastrin stimulates mucosal growth of much of the gastrointestinal tract and has also been implicated in promoting growth of colonic tumors, but its role in colorectal carcinogenesis remains controversial. This study determined fasting serum gastrin levels before and after surgery for colorectal cancer (CRC) and the relationship to the clinical stage of the disease to investigate it possible prognostic role. PATIENTS AND METHODS: Fasting radioimmunoassay gastrin, CA 19-9, and CEA levels were measured before and after surgery for CRC. Helicobacter pylori status was also assessed since it causes significant hypergastrinemia. RESULTS: Mean fasting plasma gastrin level was significantly higher in CRC patients than in controls before surgery but not 59 days after surgery. Mean CEA and CA 19-9 levels were significantly higher in patients with CRC before surgery than after tumor resection. There was a significant positive correlation between the plasma gastrin, CEA, and CA 19-9 levels and the CRC stage (Dukes' classification). CONCLUSION: The significance of gastrin as a marker for diagnosis or prognostic purposes in colorectal cancer needs to be further examined. PMID- 12548419 TI - Studies on the immunogenicity of hCEA in a transgenic mouse model. AB - BACKGROUND AND AIMS: Immunization protocols in mice have shown that the tumor associated antigen hCEA could be a target for active immunization; however, human CEA is foreign to mice. Success may depend in part on a simple anti-xenoresponse. Using hCEA-transfected syngeneic tumor cells in hCEA-transgenic mice should bypass this problem and allow testing for new vaccination strategies. MATERIALS AND METHODS: We established a hCEA transgenic model of the haplotype H2(d), which may differ from other haplotypes in cytokine production and in effectiveness of antigen presentation, and tested two vaccination protocols in wild-type and transgenic mice. RESULTS: Syngeneic wild-type mice built up an immune response with high antibody titers; only 65% of animals developed solid tumors after tumor challenge. In contrast, hCEA-transgenic mice developed no antibody response and accepted the tumor in more than 90% of cases, thus demonstrating the role of human CEA as a foreign antigen. Accordingly, active immunization using tumor lysate or lymphocytes loaded with hCEA resulted in a CTL response and tumor rejection in up to 80% of wild-type mice. hCEA-transgenic mice could be induced with both immunization protocols to build up a CTL response, although the number of CTL were much lower and the cytotoxic response weaker than in wild-type mice. In vivo hCEA-transgenic mice rejected hCEA-positive tumors only after immunization with the tumor lysate in about 60% whereas there was no rejection of tumors after immunization with the human hCEA-loaded autologous lymphocytes. CONCLUSION: The findings clearly show the importance of transgenic models when testing the effects of immunization towards human tumor associated antigens such as hCEA because results differ in wild-type and transgenic mice. PMID- 12548421 TI - Clinical markers of hypoxia and other predictive factors of survival in conservative therapy of squamous-cell carcinoma of the esophagus. AB - BACKGROUND AND AIMS: Carcinoma of the esophagus is an aggressive cancer with a high failure rate after combined local and systemic treatment modalities. One of the factors that could influence the high rate of locoregional persistence is hypoxia. Hypoxic cancers are known to be more aggressive and less responsive to chemo- and radiotherapy. We investigated the effect of several factors on overall survival and several surrogate markers of hypoxia on survival in squamous-cell esophageal cancer. PATIENTS AND METHODS: We conducted a retrospective analysis of 41 curatively treated patients with squamous-cell esophageal cancer: 30 received combined radio- and chemotherapy (cisplatin and 5-FU) and 11 radiotherapy alone. Cox regression analysis was performed to study the effect of several factors on overall survival. RESULTS: Significantly better survival was shown only in patients who were younger, received more cycles of chemotherapy, or had more proximal tumors or less advanced T stage but not for possible clinical surrogate markers for hypoxia, such as levels of hemoglobin before and during treatment or smoking. CONCLUSION: We found no clinical evidence that hypoxia plays a role in survival with squamous-cell esophageal cancer. Number of chemotherapy cycles was, independently of age, predictive of survival. Measurements of in vivo tumor oxygenation could further help in determining the role of tumor hypoxia in esophageal cancer. PMID- 12548420 TI - Abnormal expression of E-cadherin, beta-catenin, and c-erbB-2 in advanced gastric cancer: its association with liver metastasis. AB - BACKGROUND AND AIMS: We investigated expression of E-cadherin, beta-catenin, and c-erbB-2 in gastric cancer to identify molecular factor(s) relevant to development of liver metastasis, which is a frequent cause of mortality in gastric cancer patients. PATIENTS AND METHODS: We analyzed by immunohistochemistry and compared expression patterns of E-cadherin, beta catenin, and c-erbB-2 in the tumor between 40 cases of gastric cancer (GC) without (GC-H(-)) and 16 with concurrent liver metastasis (GC-H(+)). RESULTS: Loss of E-cadherin expression in the primary tumor was found in 18% of GC-H(-) and in 19% of GC-H(+). Oncogenic beta-catenin activation, represented by its nuclear translocation, was detected in 13% of GC-H(-) and in 31% of GC-H(+). There was no statistical difference in incidence of alteration in these molecules between the two groups of patients. c-erbB-2 overexpression was more frequently observed in GC-H(+) (10/16, 63%) than in GC-H(-) (5/40, 13%) while the distribution of histological types of the tumors was similar in the two groups of patients. This overexpression was also detected in metastatic liver tumors and biopsy specimens in the ten of the former group of patients. CONCLUSION: Our results strongly suggest a role of activated c-erbB-2 in the process of liver metastasis, and an importance of detection of this overexpression in biopsy specimens to identify GC patients who are at high risk of developing liver metastasis. PMID- 12548422 TI - An unusual way to diagnose a rare case of bladder carcinoma in a patient with Crohn's disease. AB - BACKGROUND AND AIMS: Crohn's disease of unknown etiology may involve the urinary tract and may include a mass lesion on cystoscopy. PATIENTS AND RESULTS: We present an unusual case of asymptomatic urinary bladder carcinoma in a patient undergoing surgery for Crohn's ileitis. PMID- 12548423 TI - Perianal Hodgkin's lymphoma complicating Crohn's disease. AB - BACKGROUND: The perianal region is a very rare location for Hodgkin's lymphoma, and clinicians may often neglect the diagnosis in patients with inflammatory bowel disease. PATIENT PRESENTATION: We present a case of perianal Hodgkin's lymphoma in patient with Crohn's disease who was on long-term immunosuppression and whose symptoms would normally be attributed to Crohn's disease. Diagnosis was based on the morphological appearance of atypical cells in the lamina propria and the immunohistochemical profile of Reed Sternberg and Hodgkin's cells, showing co expression of CD15 and CD30. CONCLUSION: Perianal complaints in patients with inflammatory bowel disease may be a manifestation of other pathology. Hodgkin's lymphoma could be a progression in the chronically inflamed tissue in this unusual location. PMID- 12548424 TI - On a curious association of diabetes mellitus and sigmoid volvulus: a preliminary report. PMID- 12548425 TI - Angular and spectral sensitivity of fly photoreceptors. I. Integrated facet lens and rhabdomere optics. AB - Three optical components of a fly's eye determine the angular sensitivity of the photoreceptors: the light diffracting facet lens, the wave-guiding rhabdomere and the light-absorbing visual pigment in the rhabdomere. How the integrated optical system of the fly eye shapes the angular sensitivity curves is quantitatively analyzed in five steps: (1) scalar diffraction theory for low Fresnel-number lenses is applied to four different facet lenses, with diameter 10, 20, 40, and 80 micro m, respectively, assuming a constant F-number of 2.2; (2) optical waveguide theory is used to calculate waveguide modes propagating in circular cylindrical rhabdomeres with diameter 1.0, 2.0, and 4.0 micro m, respectively; (3) the excitation of waveguide modes is studied with the tip of the waveguide positioned in the focal plane as well as outside this plane; (4) the light absorption from the various propagated modes by the visual pigment in the rhabdomere is calculated as a function of the angle of the incident light wave; and (5) the angular sensitivity of the photoreceptor is obtained by normalizing the total light absorption. Four wavelengths are considered: 300, 400, 500 and 600 nm. The analysis shows that the wavelength dependency of the lens diffraction is strongly compensated by that of the waveguide modes, an effect which is further enhanced by the decrease in light absorption when the mode number increases. The angular sensitivity of fly photoreceptors is robust to defocus and largely wavelength independent for all except very slender rhabdomeres. PMID- 12548426 TI - Unusual pheromone receptor neuron responses in heliothine moth antennae derived from inter-species imaginal disc transplantation. AB - Single-cell electrophysiological recordings were obtained from olfactory receptor neurons housed in sensilla trichodea along the adult antennae arising from transplantation of the antennal imaginal discs between larval male Helicoverpa zea and Heliothis virescens. The olfactory receptor neurons from the majority of type C sensilla sampled on transplanted antennae displayed response characteristics consistent with those of the species that donated the antennae. However, some of the sensilla type C sampled in either transplant type contained olfactory receptor neurons that responded in a manner typical of the recipient species or other neurons that have not previously been found in the type C sensilla of either species. The single-cell data help to explain behavioral results showing that some transplant males do fly upwind to both species' pheromone blends, an outcome not expected based on known antennal sensory phenotypes. Our results suggest that host tissue can influence antennal olfactory receptor neuron development, and further that because of a common phylogenetic ancestry the donor tissue has the genetic capability to produce a variety of sensillar and receptor types. PMID- 12548428 TI - Assessment of food source profitability in honeybees (Apis mellifera): how does disturbance of foraging activity affect trophallactic behaviour? AB - When forager honeybees (Apis mellifera) return to the hive after a successful foraging trip, they unload the collected liquid to recipient hive mates through mouth-to-mouth contacts (trophallaxis). The speed at which the liquid is transferred (unloading rate) from donor to recipient is related to the profitability of the recently visited food source. Two main characteristics that define this profitability are the flow of solution delivered by the feeder and the time invested by the forager at the source (visit time). To investigate the effect of visit time on trophallactic behaviour, donor foragers were trained to a rate feeder that could deliver different flows of solution. We dissociated visit time and flow of solution by introducing pauses in the solution's deliverance at different moments of the foraging visit. We analysed whether timing of the non deliverance period within the visit is important for the forager's assessment of resource profitability. During the subsequent trophallactic encounter with a hive mate, unloading rate was related to the total time invested by the forager at the food source only if the ingestion process had already been started. These results together with previous ones suggest that foragers integrate an overall flow rate of solution of the feeder throughout the entire foraging visit. PMID- 12548427 TI - The neuronal targets for GABAergic reticulospinal inhibition that stops swimming in hatchling frog tadpoles. AB - In most animals locomotion can be started and stopped by specific sensory cues. We are using a simple vertebrate, the hatchling Xenopus tadpole, to study a neuronal pathway that turns off locomotion. In the tadpole, swimming stops when the head contacts solid objects or the water's surface meniscus. The primary sensory neurons are in the trigeminal ganglion and directly excite inhibitory reticulospinal neurons in the hindbrain. These project axons into the spinal cord and release GABA to inhibit spinal neurons and stop swimming. We ask whether there is specificity in the types of spinal neuron inhibited. We used single neuron recording to determine which classes of spinal neurons receive inhibition when the head skin is pressed. Ventral motoneurons and premotor interneurons involved in generating the swimming rhythm receive reliable GABAergic inhibition. More dorsal inhibitory premotor interneurons are inhibited less reliably and some are excited. Dorsal sensory pathway interneurons that start swimming following a touch to the trunk skin do not appear to receive such inhibition. There is therefore specificity in the formation of descending inhibitory connections so that more ventral neurons producing swimming are most strongly inhibited. PMID- 12548429 TI - Bumble bees alert to food with pheromone from tergal gland. AB - Foragers of Bombus terrestris are able to alert their nestmates to the presence of food sources. It has been supposed that this happens at least partially through the distribution of a pheromone inside the nest. We substantiate this claim using a behavioral test in which an alerting signal is transmitted from one colony to another by long distance air transport, so excluding all other modalities of information exchange. We then investigated the source of the pheromone and were able to show that a hexane extract from tergites V-VII of bumble bee workers elicits higher activity, like a successful forager does. Extracts from other glands, such as the mandibular, labial, hypopharyngeal, and Dufour's gland as well as extracts from other parts of the cuticle had no effect. This suggests that bumble bees possess a pheromone-producing gland, similar to the Nasanov gland in honey bees. Indeed, an extract from the honey bee Nasanov gland also proved to alert bumble bee workers, suggesting a possible homology of the glands. PMID- 12548430 TI - Head bobbing and the body movement of little egrets ( Egretta garzetta) during walking. AB - Although previous studies have indicated that head bobbing of birds is an optokinetic movement, head bobbing can also be controlled by some biomechanical constraints when it occurs during walking. In the present study, the head bobbing, center of gravity, and body movements of little egrets (Egretta garzetta) during walking were examined by determination of the position of the center of gravity using carcasses and by motion analysis of video films of wild egrets during walking. The results showed that the hold phase occurs while the center of gravity is over the supporting foot during the single support phase. In addition, the peak speed of neck extension was coincident with the peak speed of the center of gravity. These movements are similar to those of pigeons, and suggest the presence of biomechanical constraints on the pattern of head bobbing and body movements during walking. PMID- 12548431 TI - Network interactions among sensory neurons in the leech. AB - Interactions among mechanosensory neurons, sensitive to touch, pressure and nociceptive stimuli in the leech nervous system were studied in isolated ganglia and in body-wall preparations. Pairs of touch-pressure, touch-nociceptive and pressure-nociceptive neurons were tested by suprathreshold stimulation of one neuron while recording the response of the other, in both directions. Pressure and nociceptive stimulation evoked depolarizing and hyperpolarizing responses in touch cells, mediated by interneurons. The relative expression of these responses depended on the stimulus duration. One or two pressure cell spikes produced, predominantly, a depolarization of the touch cells, and increasing number of spikes evoked a hyperpolarization. Nociceptive cells produced primarily the hyperpolarization of touch cells at any stimulus duration. When touch cells were induced to fire by injection of positive current into the soma, stimulation of pressure cells inhibited touch cell activity. However, when touch cells were induced to fire by peripheral stimulation, pressure cell activation failed to inhibit touch cell firing. The results suggest that excitation of pressure and nociceptive cells would not limit the responses of touch cells to peripheral stimuli, but would inhibit the firing of touch cells evoked by their central connectivity network. PMID- 12548432 TI - Contribution of classic photoreceptors to entrainment. AB - The ability to phase shift and entrain in response to light is spared in retinally degenerate mice (rd/rd). In the present work, fewer retinally degenerate C57BL/6 mice than wildtypes entrained in dim lights, suggesting that rods and/or cones contribute toward entrainment even though they are not necessary. Thresholds for entrainment appear to be a more sensitive test of deficits in entrainment than phase shifts in response to light pulses. PMID- 12548433 TI - Aschoff's rule in retinally degenerate mice. AB - Both retinally degenerate and wildtype mice lengthened the period of their free running circadian rhythms and reduced the amount of wheel running when exposed to increasing levels of constant illumination, in accordance with Aschoff's rule. Decreased locomotor activity may contribute toward lengthening of period in bright light. However, the known effects of activity on free-running period are small compared to those obtained by changing illumination. This suggests that Aschoff's rule in mice is not dependent on changes in nonphotic input, but results from a direct effect of light on the circadian system. The sparing of Aschoff's rule in retinally degenerate mice is further evidence that circadian photoreception depends on mechanisms other than rods and cones. PMID- 12548434 TI - Ankylosing spondylitis in the pharaohs of ancient Egypt. AB - BACKGROUND: Among the pharaohs of the 18th and 19th dynasty of Old Egypt, at least three had ankylosing spondylitis: Amenhotep (Amenophis) II, Ramses II ("The Great"), and his son Merenptah. OBJECTIVE: An illustrated review is given on the radiological indications for their disease, together with the rough history of these pharaohs, the history of their tombs, of the detection of their mummies in the 19th century and of their paleopathological investigation. PMID- 12548435 TI - Simultaneous presence of neutrophil alveolitis and Ki-67 positivity of alveolar macrophages in dermato-/polymyositis and systemic sclerosis. AB - OBJECTIVE: The proportion of cell proliferation-associated nuclear Ki-67 antigen positive alveolar macrophages was compared in the bronchoalveolar lavage fluids (BAL) of patients with dermato-/polymyositis (DM-PM), scleroderma-myositis overlap syndrome (Scl-M), systemic sclerosis (SSc), and undifferentiated connective tissue disease (UCTD). PATIENTS AND METHODS: We examined 8 patients with DM-PM, 3 with Scl-M, 11 with SSc, and 10 with UCTD. The patients were selected on the basis of their interstitial lung involvement diagnosed by non invasive methods. RESULTS: Increased Ki-67 expression in alveolar macrophages was observed in three cases of SSc (27%) and in 4 of myositis (36%). In UCTD and in healthy volunteers a low level of Ki-67 expression was detected (in both groups 5%). A positive correlation was found between the percentage of the Ki-67 positive alveolar macrophages and the percentage of granulocytes in BAL fluid of the investigated patients with different systemic autoimmune diseases and healthy controls, indicating that increased local proliferation of differentiated large alveolar macrophages can be detected in cases with neutrophil alveolitis. There was also a correlation between the extent of ground glass opacity on HRCT and elevated proportion of proliferating alveolar macrophages in BAL fluid. PMID- 12548436 TI - Intra-erythrocyte deposition of growth hormone in rheumatic diseases. AB - Standard radioimmunoassay was employed to quantify matched basal intra erythrocyte growth hormone (GH) and serum GH levels from patients with osteoarthritis, diffuse idiopathic skeletal hyperostosis (DISH), gout, and Sjogren's syndrome and in a group of normal volunteers (control group). Basal intra-erythrocyte GH concentration was significantly higher ( P<0.05) than serum GH concentration in blood samples from patients with osteoarthritis and DISH but not from those with gout or Sjogren's syndrome. Regression analysis determined that basal serum GH levels did not correlate with age. While intra-erythrocyte GH levels exceeded serum GH levels in osteoarthritis and DISH, intra-erythrocyte GH was not a strong predictor of serum GH levels in osteoarthritis, DISH, or in the control group. PMID- 12548437 TI - German short musculoskeletal function assessment questionnaire (SMFA-D): comparison with the SF-36 and WOMAC in a prospective evaluation in patients with primary osteoarthritis undergoing total knee arthroplasty. AB - In a prospective clinical trial, the German short musculoskeletal function assessment (SMFA-D), the short form (SF)-36, and the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) were evaluated in 63 patients with primary osteoarthritis undergoing total knee arthroplasty. All instruments were sensitive to change, demonstrating the effect of total knee arthroplasty at 1 year follow-up. The SMFA-D effect sizes in comparable scales were bigger than in the SF-36 and similar to those of the WOMAC. Significant correlations of the SMFA D indices with the SF-36 and WOMAC scales preoperatively could be shown. After 1 year follow-up, all correlations between the SMFA-D indices and SF-36 scales were significant. In other comparison, only the correlation between the SMFA-D function index and the WOMAC function scales remained significant. The correlation of the SMFA-D function index with external validation criteria was higher than that using the other instruments. We recommend the SMFA-D for assessing change in functional status of patients with primary osteoarthritis of the knee following arthroplasty. PMID- 12548438 TI - Adaptation and validation of the Turkish version of the Rheumatoid Arthritis Quality of Life Scale. AB - OBJECTIVE: The aim of this study was to adapt the Rheumatoid Arthritis Quality of Life (RAQoL) questionnaire for use in Turkey and to test its reliability and validity. METHODS: The translation process included the recent guidelines for cross-cultural adaptation. Reliability of the Turkish RAQoL was assessed by internal consistency and test-retest reliability, internal construct validity by Rasch analysis, and external construct validity by associations with impairments, disability, and general health status. Cross-cultural validity was tested through analysis of differential item functioning (DIF) by comparison with data from the UK version of the RAQoL. RESULTS: Reliability of the adapted version was good, with high internal consistency (Cronbach's alpha 0.95 and 0.96 at times 1 and 2, respectively) and test-retest reliability (Spearman's rho 0.874). Internal construct validity was confirmed by excellent fit to the Rasch model (mean item fit 0.236, SD 1.113) and external construct validity by expected associations. The DIF for culture was found in four items. CONCLUSIONS: Adaptation of the RAQoL for use in Turkey was successful. The instrument can be used in both national and international studies for cross-cultural comparison with the UK, as long as adjustments are made for the few items displaying DIF for culture. PMID- 12548439 TI - Anti-inflammatory effects of a low arachidonic acid diet and fish oil in patients with rheumatoid arthritis. AB - BACKGROUND: Patients with rheumatoid arthritis (RA) improve on a vegetarian diet or supplementation with fish oil. We investigated the effects of both dietary measures, alone and in combination, on inflammation, fatty acid composition of erythrocyte lipids, eicosanoids, and cytokine biosynthesis in patients with RA. METHODS: Sixty-eight patients with definitive RA were matched into two groups of 34 subjects each. One group was observed for 8 months on a normal western diet (WD) and the other on an anti-inflammatory diet (AID) providing an arachidonic acid intake of less than 90 mg/day. Patients in both groups were allocated to receive placebo or fish oil capsules (30 mg/kg body weight) for 3 months in a double-blind crossover study with a 2-month washout period between treatments. Clinical examination and routine laboratory findings were evaluated every month, and erythrocyte fatty acids, eicosanoids, and cytokines were evaluated before and after each 3-month experimental period. RESULTS: Sixty patients completed the study. In AID patients, but not in WD patients, the numbers of tender and swollen joints decreased by 14% during placebo treatment. In AID patients, as compared to WD patients, fish oil led to a significant reduction in the numbers of tender (28% vs 11%) and swollen (34% vs 22%) joints (P<0.01). Compared to baseline levels, higher enrichment of eicosapentaenoic acid in erythrocyte lipids (244% vs 217%) and lower formation of leukotriene B(4) (34% vs 8%, P>0.01), 11-dehydro thromboxane B(2) (15% vs 10%, P<0.05), and prostaglandin metabolites (21% vs 16%, P<0.003) were found in AID patients, especially when fish oil was given during months 6-8 of the experiment. CONCLUSION: A diet low in arachidonic acid ameliorates clinical signs of inflammation in patients with RA and augments the beneficial effect of fish oil supplementation. PMID- 12548440 TI - Prevalence of dermatophytosis in patients with rheumatoid arthritis. AB - OBJECTIVE: The prevalence of dermatophytic infections in rheumatoid arthritis is unknown. This study investigated the prevalence of dermatophytosis in patients with rheumatoid arthritis and the relationship between sulfasalazine, low-dose methotrexate and steroid therapy. METHODS: We examined 53 consecutive patients with rheumatoid arthritis for evidence of dermatophytosis and compared them 55 with age- and sex-matched, nonimmunocompromised controls recruited from the low back pain population. Nail scrapings were obtained from the subjects, and the clinical diagnosis of dermatophytosis was confirmed with a potassium hydroxide preparation. RESULTS: In 32% of the rheumatoid arthritis population we found dermatophytosis, compared with 16% of the control group, although statistical significant was only borderline. Tinea pedis was the most frequent type of dermatophytosis in both groups. The prevalence of dermatophytosis in patients receiving sulfasalazine, low-dose methotrexate, and steroid therapy was not found to be significantly increased. CONCLUSIONS: This study shows a slightly higher prevalence of dermatophytosis in rheumatoid arthritis population than in controls. Sulfasalazine, low-dose methotrexate, and steroid therapy had no effect on the prevalence of dermatophytic infections in patients with rheumatoid arthritis. PMID- 12548441 TI - Idiopathic portal hypertension in a systemic sclerosis patient heterozygous for factor V Leiden mutation. AB - Here we present a rare case of systemic sclerosis (SSc) with idiopathic portal hypertension (IPH) having factor V Leiden mutation, a well-known genetic risk factor for various venous thromboses. A 53-year-old SSc patient showing huge esophageal varices and splenomegaly without liver cirrhosis was diagnosed with IPH. As heterozygous Leiden mutation was detected, some coagulation abnormality and resultant formation of microthrombi in the branches of portal vein were suggested as a cause of IPH in this case. This is the first report showing the possible association between Leiden mutation and one of the complications of SSc. PMID- 12548442 TI - Liaison between rheumatoid arthritis and ulcerative colitis. PMID- 12548443 TI - Accuracy of image fusion of normal upper abdominal organs visualized with PET/CT. AB - Although PET/CT scanners have the potential for precise fused registration of structures visualized on both PET and CT, physiological motion during the acquisition of both studies may alter the appearance of organ shape, size or location. The degree of possible mismatch in abdominal organ size and position between PET and CT has not been evaluated. The aim of this study was to assess the consistency in location and measured size of upper abdominal organs with PET and CT using a combined PET/CT system. Forty-six consecutive inpatients who underwent clinical PET/CT scans for suspected cancer were evaluated. CT and PET images attenuation corrected by both CT and germanium-68 transmission scans were obtained, and we separately determined the location of the top and bottom (height), anterior and posterior margins (thickness), and right and left margins (width) for each organ, including liver, spleen, and bilateral kidneys, using CT and both sets of PET images. Differences between the two modalities in terms of location and measured organ size were investigated. In the upper margin of the liver and lower margin of the spleen, more than 10% of the cases showed a larger discrepancy (>20 mm) between CT-based and Ge-corrected PET-based measurements, although the differences in the positions of the edges were less than 10 mm in most cases. The center of the liver tended to be located cephalad and to the right of the body, and that of the spleen tended to be cephalad and posterior on PET, as compared with CT. Moreover, the center of both kidneys tended to be seen cephalad, posterior, and to the right on PET. The liver appeared slightly larger on PET than CT in thickness (CT vs CT-corrected PET vs Ge-corrected PET = 156 mm vs 162 mm vs 162 mm) and width (186 mm vs 189 mm vs 188 mm). By contrast, the spleen appeared slightly smaller on PET than CT in height (84 mm vs 77 mm vs 80 mm) and width (85 mm vs 81 mm vs 80 mm). A similar tendency was observed in the left kidney (105 mm vs 100 mm vs 99 mm in height, and 64 mm vs 59 mm vs 58 mm in width) and the right kidney (99 mm vs 93 mm vs 93 mm in height, and 64 mm vs 59 mm vs 60 mm in width). These differences between the two modalities were statistically significant ( P<0.05). In conclusion, minor mismatches in location and organ size were found to exist between CT and PET images, in part due to physiological motion. Although these differences could potentially affect the quality of the image registrations, they were generally of a modest nature. PMID- 12548444 TI - Intra- and extra-articular anterior cruciate ligament reconstruction utilizing autogeneous semitendinosus and gracilis tendons: 5-year clinical results. AB - This prospective study examined 50 patients who underwent ACL surgery using hamstring tendons with a modification including intra- and extra-articular reconstruction. All patients were athletes competing at a high level in various sports. Full return to sports was allowed at 4 months. IKDC score and KT-2000 were used for assessing clinical outcome at a mean of 6.4 years (5-7 years) Resumption of sport, Tegner activity score, and isokinetic test were also used. The IKDC score showed 92% of normal or nearly normal knees. Of the 50 patients 48 regained full extension, and only two had extension deficit between 0 degrees and 3 degrees. KT evaluation was less than 3 mm in 38 cases (76%), 3-5 mm in 9 (18%), and more than 5 mm in 3 (6%). In 90% of cases the patient resumed sport at the same level. The mean Tegner activity score was 8.1 (5-10). The isokinetic test showed no deficit for hamstring and quadriceps muscles. Removal of staples was necessary in eight cases (16%) due to femoral lateral bursitis. Acute reconstruction had significantly better clinical assessment of abnormal laxity and KT value. Men had significantly better results then women. This technique demonstrated a high reliability, low morbidity, low functional deficit and fast recovery using hamstring grafts. PMID- 12548445 TI - Tendon-to-bone tunnel healing in a rabbit model: the effect of periosteum augmentation at the tendon-to-bone interface. AB - We examined whether periosteum enhances the healing process of a tendon in a bone tunnel and leads to better biomechanical fixation in a shorter period of time. The effect of the periosteum-wrapped tendon on tendon-to-bone healing was analyzed in 20 New Zealand white rabbits. The animals were divided into two groups, a periosteum-wrapped group and a control group. The two legs were operated on in the same manner. The long digital extensor tendon was harvested and transplanted into the proximal tibial tunnel. One limb was transplanted with the tendon wrapped with periosteum, while the other was without periosteum. The healed tendon-bone attachment was evaluated after 3 and 6 weeks by histological examination and biomechanical testing. At all time points histological examination demonstrated more extensive bone formation around the tendon with closer apposition of new bone to the tendon in the periosteum-wrapped limb than in the control limb. Biomechanical testing demonstrated higher tendon pullout strength in the periosteum-wrapped limb at all time points, with statistically significant differences between the periosteum-wrapped limb and the control limb after 3 and 6 weeks. Histological and biomechanical data suggest superior healing at the periosteum-wrapped side. These findings demonstrate that periosteum enhances the healing process when a tendon graft is transplanted into a bone tunnel. PMID- 12548446 TI - Weight-bearing knee kinematics in subjects with two types of anterior cruciate ligament reconstructions. AB - Weight-bearing knee kinematics in patients who received two types of anterior cruciate ligament reconstruction were studied using a fluoroscopy-based three dimensional measurement technique. Eleven patients with more than 1-year follow up and good or excellent results participated in this study. Six subjects received anterior cruciate ligament reconstruction using a multiply folded semitendinosus and gracilis tendon graft, and five received combined intra- and extra-articular anterior cruciate ligament reconstruction using the iliotibial tract. The step up/down activity of normal and operated knees was recorded using lateral fluoroscopy. A customized three-dimensional contour model was created from two orthogonal views of each knee and all six degrees of freedom of knee movement were determined using a model matching technique. Both the normal and the reconstructed knees exhibited posterior condylar translation and internal tibial rotation with knee flexion, consistent with previous reports of normal kinematics. There were no statistically significant differences in the axial rotations or lateral or medial condylar anterior/posterior translations between operated and normal knees or between the two groups of operated knees. PMID- 12548447 TI - A biomechanical study of four different meniscal repair systems, comparing pull out strengths and gapping under cyclic loading. AB - Most studies comparing the biomechanical properties of different meniscal repair systems have simply investigated load to failure. Meniscal tissue is highly anisotropic, and far weaker under tension in the radial direction. Radially oriented loading to failure may not therefore be the most physiologically relevant in vitro test for repair of circumferential tears, and determining gapping across repair sites under cyclical loading at lower loads may be of greater importance. Using bovine menisci, vertical circumferential incisions were repaired using a simple vertical 2-0 PDS suture, Meniscal Arrow, Meniscal Fastener or T-Fix. Repairs were tested by simple loading to failure in a materials testing machine, and by cyclic loading between 5 and 10 N for 25 cycles. Initial gapping across the repairs was measured using a digital micrometer, and the increase in gapping under cyclic loading measured using a Differential Voltage Reluctance Transducer. The mean loads to failure for each of the repair groups were: sutures 72.7 N, Arrows 34.2 N, Fasteners 40.8 N and T-Fix 49.1 N. The load to failure was significantly greater with sutures than with Arrows or Fasteners. The mean gapping across the repairs for each of the repair groups after 25 loading cycles were: sutures 3.29 mm, Arrows 2.18 mm, Fasteners 3.99 mm and T-Fix 3.47 mm. The mean gapping was significantly less with Arrows than with Sutures, Fasteners or T-Fix. These results confirm that meniscal repair by suturing gives the highest load to failure, but show that Arrows give superior hold under lower loads, with the least gapping across repairs under cyclic loading of the four methods tested. PMID- 12548448 TI - Arthroscopy alone or in association with open cystectomy: in treatment of lateral meniscal cysts. AB - We retrospectively the results in 27 knees (26 patients) with cysts of the lateral meniscus. All patients had a meniscal tear (20 horizontal, 7 transverse) and pain over the lateral joint line with a palpable mass. The pathology was in the middle or anterior one-third of the meniscus. Arthroscopic surgery was performed in 22 knees, consisting of partial excision of meniscal tear, leaving a peripheral rim, coupled with debridement of the cyst; open cystectomy was performed in the remaining 5 knees. Average follow-up was 58 months (range 9 110). All patients but two were satisfied with the results. PMID- 12548449 TI - Transchondral drilling for osteochondritis dissecans of the medial condyle of the knee. AB - Osteochondritis dissecans is a rare condition which occurs most frequently in the medial condyle of the knee. Its prognosis remains controversial. We report our experience with transchondral drilling (TCD) for osteochondritis dissecans. The rationale for this low-morbidity procedure is to enhance the healing potential of a lesion which failed to heal after conservative treatment. This multicenter retrospective study examined 24 patients (25 medial condyles), including cases both before (juvenile osteochondritis dissecans, JOCD; n=17) and after closure of the physis (adult osteochondritis dissecans, AOCD; n=8). All patients initially underwent a prolonged conservative treatment. The mean duration of symptoms prior to surgery was 22 months in JOCD and 55 months in AOCD patients. TCD was proposed in cases of intact or almost intact articular cartilage and was initially conducted under arthrotomy and later by arthroscopy. Mean follow-up time was 11.8 years in JOCD and 6 years in AOCD, and evaluation was based on clinical examination and radiography. Excellent clinical results were only observed in the JOCD group (12/17). In the AOCD group 4 patients had a good result and the other 4 a poor result. Radiological findings matched with clinical results. Except for age, factors of poor prognosis were: fissure of the articular cartilage and anterior extension of the lesion. We do not recommend TCD for AOCD. In JOCD, failure after 12 months of conservative treatment should indicate TCD in the cases where the cartilage remains continuous. At this stage there is no indication for more aggressive procedures. PMID- 12548450 TI - Unicompartmental knee prosthesis implantation with a non-image-based navigation system: rationale, technique, case-control comparative study with a conventional instrumented implantation. AB - The accuracy of implantation is an accepted prognostic factor for the long-term survival of unicompartmental knee prostheses (UKP). We developed a non-image guided navigation system for UKP implantation without any extramedullary or intramedullary guiding device. The 30 patients operated on with the navigation system (group A) were matched to 30 patients operated on with the conventional technique (group B) using age, sex, body mass index, preoperative coronal mechanical femorotibial angle, and severity of the preoperative degenerative changes. All patients had a complete radiological examination in the first 3 months after the index procedure, with anteroposterior and lateral plain knee radiographs and anteroposterior and lateral long leg radiographs. Coronal femorotibial mechanical angle and both coronal and sagittal orientations of the femoral and tibial components were measured. There were no significant differences in the mean numerical values of all measured angles except for the sagittal orientation of the tibial component, with a significant excessive posterior tibial slope in group B. There was a significant increase in the rate of prostheses implanted in the desired angular range for all criteria except the coronal mechanical femorotibial angle in group A. An optimal implantation with all optimal items was obtained by 18 cases in group A and 6 cases in group B. Navigated implantation of a UKP with the used, non-image-based system improved the accuracy of the radiological implantation without any significant inconvenience and with little change in the conventional operative technique. The only inconvenience was a 20-min longer operative time. This improvement could be related to a longer survival of such implanted prostheses. PMID- 12548451 TI - Improvement in technique for arthroscopic ankle fusion: results in 15 patients. AB - We retrospectively assessed time until consolidation, complications, and functional results according to Morgan from the clinical charts and radiographs of 15 arthroscopic ankle fusions. In 11 patients unilateral distraction and crossed screw placement over the fusion area through tibia and fibula were used (group A); in 4 patients a technique of bilateral distraction and parallel screw placement from the dorsal side of the tibia into the neck of the talus was used (group B). In group A there were two cases of insufficient compression at the arthrodesis site, three cases of suboptimal compression, and five cases of malposition of the screws. In all cases in group B good compression and fixation was achieved, and no case of malpositioning of screws occurred. There was nonunion in 3 of 11 patients in group A and in none of the four patients in group B. Time until fusion was 23.3 in group A and 12.5 weeks in group B. Functional results were better in group B. The initial experiences with our technique of bilateral distraction and parallel screw placement are therefore promising. Screw placement is easier and optimal compression and fixation are achieved. We feel that this technique should be considered when performing an arthroscopic ankle fusion. PMID- 12548452 TI - Technique of synovial biopsy of metacarpophalangeal joints using the needle arthroscope. AB - We demonstrate the technique, advantages, and disadvantages of metacarpophalangeal joint examination with needle arthroscope. We evaluated our experience from biopsies of 10 metacarpophalangeal joints of eight rheumatoid women aged 41-45 years. The procedures were performed using a 1-mm needle arthroscope. The synovium biopsy was taken with a microforceps. The procedure was performed under local anesthesia. The tight tension of the joint and traction of the finger is necessary for good visualization, but despite this visibility can be difficult. Needle biopsy is a useful method for the early diagnosis of rheumatoid arthritis. PMID- 12548453 TI - Levels of analysis in knee surgery. PMID- 12548457 TI - [Epidemiological data of patients with sudden hearing loss -- a retrospective study over a period of three years]. AB - In this retrospective study data of 257 patients (11- 6 years; 52 % male, 48 % female) admitted during the last three years for treatment of sudden unilateral hearing loss are statistically evaluated. No correlation with coexisting disease like hypertension, coronary heart disease, hypercholesteremia, arthropathy of the cervical spine, diabetes, thyroid disorders or nicotine abuse were evident. In 194 patients (75 %) total remission was achieved after rheological therapy. Patients additionally treated with steroids had a significant better outcome. Younger patients had higher remission rates. The best prognosis was found for patients with hearing loss in the lower frequency range. There were no differences in remission rates for patients with preexisting sensorineural hearing loss or chronic otitis media. Also patients suffering from recurrent episodes of sudden hearing loss had a similar outcome also. PMID- 12548458 TI - [Chronic tinnitus and allergies -- are they linked?]. AB - BACKGROUND: Some neuro-otologic disorders are suspected to have a higher prevalence of allergies than is found in the population. This study aimed to determine the prevalence of allergies in patients with chronic tinnitus. METHODS AND PATIENTS: 102 tinnitus patients were mailed a survey containing a list of allergies, and asked to quote any allergy they were afflicted with. Additionally, they had to specify whether the diagnosis has been medically confirmed or not. Findings were compared to those from an age- and gender-matched control group. RESULTS: 32 % of tinnitus subjects (CI (95 %) = 42 - 24 %), but only 21 % of controls (CI (95 %) = 29 % - 13 %) had at least one medically diagnosed allergy. Regarding all (ie., diagnosed and non-diagnosed) allergies, 49 % of tinnitus subjects (CI (95 %) : 39 -59 %) versus 46 % (CI (95 %) : 37 -57 %) of controls stated to be afflicted with at least one allergy. CONCLUSIONS: Our findings suggest that the frequency of allergies in tinnitus patients is essentially equal to the allergy prevalence in the population as revealed from recent epidemiologic studies. We conclude that there is no increased risk for allergies in chronic tinnitus. The lower rate of medically diagnosed allergies in non-tinnitus subjects probably indicates that mild allergies are less frequently diagnosed in the group of otherwise healthy persons. PMID- 12548459 TI - [How do metallic middle ear implants behave in the MRI?]. AB - BACKGROUND: Magnetic resonance imaging (MRI) has gained in frequency and importance as a diagnostic procedure. In respect to the close anatomical relationship in the temporal bone it is necessary to know whether it is hazardous to patients with metallic middle ear implants regarding displacement and rise in temperature. For the MR image quality artefacts caused by metallic prostheses should be low. METHODS: Four different stapes prostheses made from titanium, gold, teflon/platinum and teflon/steel, a titanium total ossicular reconstruction prosthesis (TORP) and two ventilation tubes (made from titanium and gold) were tested in a 1.5 Tesla MRI machine regarding their displacement. All objects were first placed in a petri dish, then suspended from a thread and finally immersed in a dish filled with Gadolinium. Temperature changes of the implants were recorded by a pyrometer. RESULTS: None of the implants moved when they were placed in the petri dish or suspended from the thread. On the water surface the teflon/platinum and the teflon/steel pistons adjusted their direction with their axis longitudinally to the MRI scanner opening and the teflon/steel piston floated towards the MRI-machine when put close enough to the scanner opening. No rise in temperature was recorded. All implants showed as little artefacts that would still make an evaluation of the surrounding tissue possible. CONCLUSION: Patients with any of the metallic middle ear implants that were examined in this study may undergo MRI-investigations without significant adverse effects. PMID- 12548460 TI - [Cutaneous malignant melanoma of the head and neck with intermediate tumor thickness: the role of elective lymph node dissection for clinical stage I]. AB - BACKGROUND: At present a tendency towards a more limited surgery in the treatment of clinical stage I and II melanoma exists. The controversy of elective lymph node dissections (ELND) in stage-I-melanoma with intermediate tumor thickness continues to be discussed. The sentinel lymph node biopsy may provide improved staging accuracy. METHODS: It was the aim of this study to examine whether extent of surgical treatment particularly the ELND in case of intermediate tumor thickness has prognostic significance. Metastatic pattern of these malignant melanoma were analyzed to form a basic for routine sentinel lymph node biopsy. RESULTS: No prognostic difference could be shown in clinical stage I melanoma with intermediate tumor thickness between "local excision" (no lymph node dissection) or "wait and see" (delayed lymph node dissection because of clinical suspicion) respectively and "local excision with elective neck dissection". The 5 year survival was 67 %, 67 % and 69 %. It ran to 53 % in case of therapeutical lymph node dissection as primary therapy. Occult nodal metastases occur in 15 % and "skip"-metastases in 6 %. CONCLUSIONS: No therapeutic value for ELND in clinical stage I malignant melanoma of the head and neck with intermediate tumor thickness could be shown. The low incidence of occult nodal metastases and "skip" metastases represents the basic for sentinel lymph node biopsy. PMID- 12548461 TI - [First results on the influence of polymorphisms at glutathione S-transferase, cytochrome P450, and tumor necrosis factor gene loci on the development of multiple head and neck cancer]. AB - BACKGROUND: The polymorphic gene loci of the detoxifying enzymes GST and CYP are susceptibility factors for single head and neck cancer. In patients with multiple cancer development a cumulation of genetic risk factors can be assumed. METHODS: Accordingly, we have examined genotype frequencies at GSTM1, GSTT1, GSTM3, GSTP1, CYP2D6, CYP1A1, CYP2E1, and TNF loci, in patients with single and multiple squamous cell carcinoma (SCC) of the head and neck, and controls. RESULTS: GSTT1 0 was increased in multiple SCC patients (36.8 %) compared with single SCC patients (21.2 %) and controls (22.2 %). GSTM3 AA was more frequent in the single SCC patients (76.6 %) and multiple SCC patients (82.1 %) compared with controls (66.5 %) but lacked significance. The TNF allele B3 was significantly increased in the single SCC group (B3-allele: 22.5 %, B3/B3 : 10.8 %) and showed a further cumulation in the multiple SCC group (B3 : 30.3 %, B3/B3 : 15.8 %). CONCLUSIONS: Patients suffering from multiple SCC development showed a cumulation of high risk genotypes at GST and TNF gene loci. Specific genetic variations could not be determined. PMID- 12548462 TI - [Review part I: basic mechanisms of cutaneous woundhealing]. AB - Cutaneous woundhealing is a complex system integrating cellular and extracellular components. It is basically divided in 3 stages: First the physiological exsudative inflammation, followed by the proliferation of granulation tissue and finally the collagen-synthesis, scarring and remodelling. Although these systems seem to be apt for skin repair the final product consists of low quality tissue. Disorders of this fine balance turns acute physiological woundhealing into chronic woundhealing. This is to be dealt with in the second part of this review. PMID- 12548463 TI - [Review part II: disorders in cutaneous woundhealing]. PMID- 12548464 TI - [Gene expression, gene regulation and gene transfer in the inner ear]. PMID- 12548465 TI - [Bonn statement on the future of hearing aid supply]. PMID- 12548466 TI - [Drug treatment for vertigo]. AB - The approach to drug treatment of vertigo is almost exclusively symptomatic. There are 3 major goals for drug treatment of vertigo: to eliminate the hallucination of motion, to reduce the accompanying neurovegetative and psychoaffective signs (nausea, vomiting, anxiety), and to enhance the process of "vestibular compensation" to allow the brain to find a new sensory equilibrium in spite of the vestibular lesion. Three different types of vertigo drug treatment must be distinguished: the treatment of acute vertigo attacks, the treatment of chronic vertigo, and the treatment of patients with Meniere's disease to avoid vertigo attacks. Furthermore, this paper deals with the treatment of motion sickness and of the postoperative nausea and vomiting. PMID- 12548467 TI - [Change in corneal refractive index in refractive surgery]. PMID- 12548468 TI - [Pitfalls of IOL power prediction after photorefractive keratectomy for high myopia -- case report, practical recommendations and literature review]. AB - BACKGROUND AND PURPOSE: Published experience with eyes after keratorefractive correction of myopia indicates that insertion of the average keratometric readings into standard IOL power predictive formulas will frequently result in substantial undercorrection and postoperative hyperopic refraction or anisometropia after cataract surgery depending on the amount of myopia corrected previously. The purpose of this paper is to discuss the accentuated differences of various approaches to minimize IOL power miscalculations by describing a case report of a patient with excessive myopia as well as a review of the literature. PATIENT AND METHODS: A 50-year old lady presented for cataract surgery on her left eye after having PRK seven years ago elsewhere (refraction - 25.5 - 3.0/20 degrees, central keratometric power 43.0 diopters [D]). Central power before cataract extraction was measured to be 35.5 D (Zeiss Keratometer) and 36.5 D (TMS 1 topography analysis) and refraction was - 3.0 D (before onset of index myopia). Orbscan slit scanning topography analysis displayed an anterior surface power of 36.8 D and a posterior surface power of - 9.3 D. Total axial length was 31.93 mm (optical biometry using Zeiss IOL-Master). The contralateral eye after PRK suffering from a comparable excessive myopia had required an exchange of the IOL implant because of intolerable anisohyperopia of + 6.0 D after primary cataract extraction elsewhere. RESULTS: Corrected corneal power values for the left eye were calculated as follows: (1) spherical equivalent (SEQ) change at spectacle plane 19.0 D, (2) SEQ change at corneal plane 26.2 D, (3) separate consideration of anterior and posterior curvature 27.5 D, (4) consideration of the IOL power misprediction on the fellow eye 29.5 D, (5) subtraction of 24 % of the SEQ change at the spectacle plane from the actually measured keratometry value 29.7 D, (6) clinical estimate from regression analysis performed earlier 30.5 D, (7) change of anterior surface power 34.5 D. Deciding for a presumably "real" corneal power of 28.0 D the Haigis formula was used to aim for - 2.0 D since the patient preferred to read uncorrected. Thus, a 21.0 D IOL was implanted uneventfully in the capsular bag. The stable refraction postoperatively was - 3.5 - 1.0/20 degrees and visual acuity increased to 20/30. Therefore, the "real" power of that cornea must have been around 30 D. CONCLUSIONS: After corneal refractive surgery, various techniques to determine the current corneal power should be compared and the value around which results tend to cluster should be relied on to avoid hyperopia after cataract surgery with lens implantation. In those cases where keratometry and refraction before PRK/LASIK are available, the gold standard is still to subtract the change of the SEQ at the corneal plane from the preoperative central keratometric power, although in the present case report the subtraction of 24 % of the SEQ change at the spectacle plane from the measured corneal power value seemed to produce the best result. Pure subtraction of the SEQ change at the spectacle plane from the corneal power value before refractive surgery has to be avoided in eyes with excessive myopia. The most reliable corrected power value should be inserted in more than one modern third-generation formula (such as Haigis, Hoffer Q, Holladay 2, SRK/T) and the highest power IOL should be implanted. In all instances, the cataract surgeon has to make sure that the corrected K-reading is not wrongly re-converted within the IOL power calculation formula used. PMID- 12548469 TI - [Do prisms according to Hans-Joachim Haase influence ocular prevalence?]. AB - BACKGROUND: Ocular prevalence is defined as an unequal weighting of the eyes in the directional perception of stereo objects. Opinions differ as to the cause and relevance of ocular prevalence. Hans-Joachim Haase suggested that ocular prevalence is due to fixation disparity, brought about by incomplete compensation of heterophoria. He further suggested that prismatic spectacles determined by his "measuring and correcting methodology" (MKH) could restore bicentral fixation and thus establish a perceptual balance between both eyes. METHODS: We examined 10 non-strabismic subjects with a visual acuity of > or = 1.0 in both eyes. It turned out that all 10 had a "fixation disparity type II", characterised according to Haase by a "disparate retinal correspondence". All subjects underwent the automatic Freiburg Ocular Prevalence Test, without and with MKH prisms. In addition we examined ocular prevalence under forced vergence and compared ocular prevalence with stereoacuity. RESULTS: Spontaneous ocular prevalence ranged between 1 and 69 %. Averaged over all 10 subjects, ocular prevalence without and with the MKH prisms were not significantly different. Statistical evaluation of single subjects revealed only in one of the 10 a significant difference (Bonferroni-corrected p = 0.001). In the subgroup of 5 subjects who underwent forced vergence, ocular prevalence remained unaltered between 0 and 18 Delta base out. The stereoscopic threshold of all 10 subjects ranged between 1.5 and 14.5 arcsec. There was no correlation between ocular prevalence and stereoscopic threshold (r = - 0.2, p = 0.5). CONCLUSION: Our results indicate that ocular prevalence is largely independent of phoria correction and vergence stress. The excellent stereoacuity of all subjects suggests that ocular prevalence is abandoned for the sake of optimal resolution when very small differences in depth have to be judged. PMID- 12548470 TI - [Flicker comparison of optic disc photographs: sensitivity and specificity]. AB - BACKGROUND: Examination and documentation of the optic nerve head are essential in monitoring glaucoma patients. Even minor changes in optic nerve head morphology can be visualised using the so-called flicker test: Two optic nerve head photographs, taken at consecutive examinations, are superimposed by projection. When occluding the pictures in a rapid alternating fashion, changes in optic nerve head morphology appear as motion. In this study, we evaluated sensitivity and specificity of the flicker test. METHOD: A set of 33 pairs of serial optic disc slides was used as gold standard. These 33 pairs had been classified earlier by 3 independent groups of experts. 23 had been classified as "no change over time", 10 had been classified as "change". All 33 pairs were now evaluated by flicker comparison in a masked fashion. RESULTS: Flicker comparison usually took 1 minute per pair of slides. Sensitivity was 90 %, specificity was 65 %. CONCLUSION: The sensitivity was reasonably high. The moderate specificity was due to some cases showing "change" with the flicker comparison which might have been overlooked by the expert groups. Flicker comparison is an easy, fast and reliable technique to evaluate pairs of consecutive optic disc photographs. PMID- 12548471 TI - [Ophthalmology "Made in Switzerland" -- Swiss papers listed in Medline]. AB - BACKGROUND: Publications are one of the most important tools to measure scientific production in a particular sphere. Hence, the aim of this study was to identify the quantitative and (through Journal Impact Factor) qualitative amount of ophthalmological articles coming from Switzerland. MATERIALS AND METHODS: With a specifically developed software we searched 70 ophthalmological Journals indexed by Medline for articles from Switzerland. RESULTS: Not counting the publications related to the annual meeting of the Swiss Ophthalmological Society, the amount of Swiss papers in ophthalmological journals indexed by Medline, increased between 1988 and 1999 by a factor of four. In proportion to the global amount of ophthalmological publications we calculated an increase by a factor of 2.6. The amount of publications grew particularly during the analysed time period in journals with high impact factors. In 1999 a total of 1.8 % of the indexed ophthalmological articles originated from Swiss research institutes and hospitals. The average amount of authors of Swiss publications increased, thereby following an international trend. CONCLUSIONS: The increased amount of publications may rely on the growing role of Swiss research in ophthalmology as well as on a broadened and deepened international orientation of Swiss ophthalmology. The increased number of authors can be explained by the growing complexity and interdisciplinary nature of research projects. Both, the amount of publications and the number of authors, can be linked to the "publish or perish" paradigm. PMID- 12548472 TI - [An outline for the standard graphical documentation of pathological macular findings]. AB - BACKGROUND: Accurate documentation and follow-up of pathological findings of the macula require graphical methods for clinical evaluation. METHODS: Symbols describing and identifying different findings of the macula were evolved. A pattern for the standardised representation of the macular region -- with size and distribution of different areas -- was also developed. RESULTS: Use of different symbols supported by a standard pattern of the macula as background simplifies documentation of pathological macular findings. CONCLUSIONS: Graphical documentation of pathological macular findings allows simple, inexpensive and standardised assessment of such findings. PMID- 12548473 TI - [Retrobulbar irradiation for Graves' ophthalmopathy -- long-term results]. AB - BACKGROUND: Significance of retrobulbar irradiation in patients suffering form Graves' ophthalmopathy, though established since almost one century, is subject of scientific debate. The present study investigated the effect of retrobulbar irradiation using a standardized protocol focussing on long term results. PATIENTS AND METHODS: Between 1981 and 1997, 104 patients treated by retrobulbar irradiation (10 to 20 Gray) due to Graves' disease. Twenty-nine of these underwent irradiation as sole treatment (mean follow-up 57 months), while in the remaining 75, it was combined with a systemic steroid treatment (mean follow-up 40 months). Patients were evaluated regarding proptosis, intraocular pressure, lid signs, motility as well as subjective assessment of double vision and retrobulbar pain. RESULTS: While proptosis remained unchanged, lid signs, chemosis and intraocular pressure showed slight and statistically significant improvement. However, these findings were considered to be clinically insignificant. Retrobulbar pain was improved in 75 % of patients. 25 % of patients showed improved motility, 75 % remained stable, and in none of them was there a deterioration of ductions. Results proved stable even in long-term follow up. 25 % per cent of patients underwent a surgical procedure one year after radiotherapy. CONCLUSIONS: In our series, we could not demonstrate an additional benefit of systemic steroids when combined with retrobulbar irradiation. Up to sixteen years after treatment, no treatment-related adverse reaction was seen. We found a remarkable improvement in ocular motility. This holds even more true in comparison to the natural course of the condition. Retrobulbar irradiation seems to shorten the duration of the disease, thus allowing earlier performance of eventual rehabilitative surgery. PMID- 12548474 TI - [Macular hole surgery: experience with autologous platelet concentrate and indocyanine green-assisted internal limiting membrane peeling]. AB - BACKGROUND: To report on our clinical experience with autologous platelet concentrate and indocyanine green(ICG)-assisted internal limiting membrane (ILM) peeling in macular hole surgery. PATIENTS AND METHODS: Standard 3-port vitrectomy was performed in 107 eyes of 101 patients (m: f = 27 : 74; mean age 60 +/- 9, range 30 - 80 years) with idiopathic macular hole stages II - IV. After fluid/air exchange, autologous platelet concentrate was applied (1.9 +/- 0,1 x 10(8) thrombocytes). ILM peeling, which was preceded by ICG staining in 19 eyes, was performed in 34 patients. RESULTS: After one procedure, anatomic success (hole closure) could be achieved in 85 % (n = 68), 75 % (n = 27) and 100 % (n = 3) of the eyes with stage II, III and IV holes, respectively. The mean visual acuity improved by 1 line. The overall initial closure rate of 82 % could be further enhanced to 96 % with a second procedure. In eyes pretreated with ICG, an initial rate of hole closure in 94 % and an improvement of visual acuity by 2 lines was observed. CONCLUSION: Autologous platelet concentrate appears to be a safe and reliable adjunct to improve the anatomical outcome of conventional macular hole surgery. Functional results can be further enhanced by ICG-assisted ILM peeling PMID- 12548475 TI - [Primary lipid keratopathy]. AB - BACKGROUND: A 73-year old man presented with a bilateral reduction of visual acuity which had slowly occurred over the last 20 years. Ophthalmologic examination revealed the presence of stromal opacifications which were denser in the central cornea associated with cataractous changes. His general history was remarkable for a status post polio infection in the early childhood and a medically treated arterial hypertension. We performed a triple procedure and histologic examination of the corneal button revealed the presence of a lipid keratopathy. CONCLUSION: Lipid keratopathy is mostly caused by corneal vascularization or chronic inflammation or is a consequence of systemic disorders of lipid metabolisms. Our patient did not exhibit these underlying disorders and therefore we made the diagnosis of a primary lipid keratopathy. The underlying corneal pathomechanisms are incompletely understood but they should be limited to the corneal keratocytes. PMID- 12548476 TI - [Ocular angiostrongylus cantonensis in a female Vietnamese patient: case report]. AB - BACKGROUND: Angiostrongylus cantonensis is largely responsible for human cases of eosinophilic meningitis and meningoencephalitis in the South Pacific and South East Asia regions. Ocular manifestations of angiostrongylus are relatively rare. We report intraocular infestation and successful removal of a worm from the anterior chamber of a patient in Vietnam. PATIENT AND METHODS: A 3-year old patient was referred to the eye hospital in Ho Chi Minh city, Vietnam, in December 2001 with clinical signs of acute iridocyclitis. The child and his family lived near the Mekong river. The medical history reported by the parents said that the girl had eaten raw snails two weeks before. After a week she developed high fever and for 3 days the right eye was red and painful. Her 8-year old brother was proven to have eosinophilic meningitis at the same time. RESULTS: At first examination, biomicroscopically a 10 mm long and actively mobile, white, round worm was detected in the anterior chamber. Marked hyphema was also present. Routine blood examination showed pronounced eosinophilia. Preoperative treatment with systemic steroids and constriction of the pupil with 1 % pilocarpine were initiated. After immobilising the worm by use of a viscoelastic agent in the anterior chamber, it was successfully removed via a clear-corneal incision. The worm was identified as a male Angiostrongylus cantonensis. Postoperative ophthalmoscopic examination showed an inactive organised cyst at the optic disc border. The child was discharged from the hospital after a week with a visual acuity of 0.6. CONCLUSION: Parasitic infestation is a major health problem in tropical countries. This is the first description of ocular infestation of Angiostrongylus cantonensis in children. No antiparasitic drugs are yet available to treat ocular involvement, thus therapeutic success depends upon early and complete surgical removal of the parasite. PMID- 12548477 TI - [V: p values: what they do tell and what they don't...]. AB - Both an extensive data description and an explicit assessment of a study result's statistical significance should be presented in the result section of a clinical trial report. Whereas the description illustrates the order and clinical relevance of the study findings, the statistical significance describes its generalizability to patients not included in the clinical trial: Despite the random recruitment of patients into a trial, the study results may fail to represent clinical reality (for example the trial might show falsely positive efficacy findings, whereas in "clinical reality" efficacy appears rather limited). A p value measures the statistical significance of a study result -- the smaller the p value turns out, the fewer statistical evidence for "random" findings. The clinical relevance of study findings can be measured by median differences or absolute and relative risks, for example, between two therapeutic strategies. Compact table structures for the simultaneous representation of clinical relevance and statistical significance in a paper's result and discussion section are proposed. PMID- 12548478 TI - [Model project on coding accuracy]. AB - First experiences with the external evaluation of coding accuracy in view of the German DRG-System are reported. 387 randomised inpatient cases of three departments of a municipal hospital were evaluated. 1.648 diagnosis codes and 946 procedure codes were evaluated with complete clinical data. Before and after correction by the reviewers the cases were grouped (AR-DRG 4.1) and the casemix index of the sample was calculated. 45.9% to 56.7% of primary diagnosis in the department samples were rated as correct. 25.2% to 37.5% of secondary diagnosis were rated as correct, 8.3% to 14.2% were corrected and 49.2% to 60.5% were rated as not relevant with regard to the German coding standards. 7.2% to 22.7% of secondary diagnosis had to be completed in the data. Evaluation of procedure codes resulted in 54.2% to 65.7% accepted codes, 5.9% to 12.1% corrected codes and 23.1% to 39.9% not accepted with regard to the German coding standards. 30.8% to 37.0% of procedure codes had to be completed in the data. After review, remarkable shift in DRGs was seen and casemix index increased 6.9% in average (0.25-12.1%). General and department-specific implications for improvement of coding accuracy could be evaluated. Consequences of potential coding errors in a prospective payment system on DRG basis were seen under conditions of daily hospital practice. External evaluation of coding accuracy used in this study could be the methodological basis for further investigations on this topic. PMID- 12548479 TI - [Fair hospital comparisons -- does the method contracted in Germany enable unbiased results?]. AB - AIM: To evaluate the method for comparison of average length of stay in hospitals as defined in a contract between German health insurance companies and the "Deutsche Krankenhausgesellschaft" (German Hospital Association). METHOD: Simulation study executing the algorithm agreed upon in different scenarios, which varied the number of hospitals to be compared, the dispersion of diagnostic specialization over hospitals, and the distribution of hospitals' sizes. Scenarios were constructed to realistically reflect the situation in German inpatient treatment of mentally ill patients. RESULTS: By fixing casemix adjustments only on diagnoses of patients and by doing so aggregated on the level of hospitals, the method for comparison yielded artificial differences between hospitals even in a situation where each patient is treated with exactly the same amount of resource allocation (given the individual needs defined for all scenarios). Results of artificial differences were heavily biased against or in favour of the reference hospitals according to the specific condition of the scenario parameters. CONCLUSION: The contracted method is not capable of achieving fair hospital comparisons, at least not for psychiatric hospitals. PMID- 12548480 TI - [Guidelines for the sociomedical assessment of performance in patients suffering from discopathy or associated diseases]. AB - The following guidelines were developed for the medical assessment services of the German Federal Insurance Institute for Salaried Employees (BfA). Starting from day-to-day practice, criteria and attributes to guide decisions for a systemisation of the sociomedical assessment of performance in discopathy or associated diseases were compiled. The guidelines aim at standardising the sociomedical assessment of performance and help to make the decision-making process more transparent -- e. g. for the assessment of applications for decreased earning capacity benefits. The guidelines summarise typical manifestations of discopathy or associated diseases and describe the necessary medical information for the sociomedical assessment of performance. Relevant assessment criteria for the medical history, clinical examination, and for diagnostic tests are illustrated. The assessment of the individual's capacity is outlined, taking occupational factors into account. Following the determination of dysfunctions the remaining abilities and disabilities, respectively, are deduced and compared with occupational demands. Finally, inferences are drawn regarding the occupational capacity of the individual. Guidelines for the sociomedical assessment of performance in coronary heart disease (CHD) have been published in Gesundheitswesen 2002, 64: 451-465. PMID- 12548481 TI - [Premature departure from nursing in Germany as a growing problem for the health care system -- a review]. AB - There is a shortage of nursing staff in almost all of the countries in the European Union. This problem is expected to increase within the next decades as a result of demographic changes. Efforts to improve training or recruiting nurses from other countries such as future EU member states are unlikely to solve the problem. Premature departure from the nursing profession occurs more frequently than in other professional fields in Germany. There is little known about the underlying reasons of this. The authors present data regarding the early departure of nursing staff from the profession. They propose that a lengthened stay in nursing could help solve the above problem. In order for specific measures to take place (to solve this), more knowledge is needed regarding the causes and circumstances surrounding the early departure of nursing staff. This is the object of the European NEXT-Study ( http://www.next-study.net ), in which research groups from 10 European countries are taking part. Starting in Autumn 2002, between 5000 and 8000 nurses will be questioned in each of the 9 countries, in this longitudinal study. The first findings will be submitted in Summer 2003. PMID- 12548482 TI - [Perceived general health and mortality]. AB - Many international studies underline the importance of the health indicator "perceived general health" (PGH). For Germany, only few Longitudinal studies are available from recent years, dealing with the question whether PGH is a predictor for overall mortality. Based on a Mortality-Follow-up Study (n = 7212) under the auspices of the Federal Institute for Population Research the importance of the indicator PGH for the prediction of the mortality experience in the general population has been analysed for the period 1984 - 1998. The age-adjusted relative risk of "less than good health" for overall mortality is 2.11 (p <0.001) for males and 2.05 (p <0.001) for females (reference category: "good/very good health"). The relative risks for "poor health" are 4.32 (p <0,001) in males and 3.07 (p <0.001) in females. An inclusion of several control variables remarkably reduces these relative risks. The results indicate that the indicator "perceived general health" is an important predictor of overall mortality in Germany. PMID- 12548484 TI - [Gottingen declaration on the education of medical students in evidence-based medicine -- on the occasion of the first teaching conference of the German Network for Evidence-based Medicine in November 2002 at Gottingen]. PMID- 12548483 TI - [Slot machine gambling behaviour in East Germany 1990-2000]. AB - TARGET: To test the hypothesis that parameters of slot machine gambling behaviour and related treatment cases have been increasing heavily in the new Lander of Germany (after reunification). METHOD: Data from the National Population Survey 2000 and from the EBIS Treatment Monitoring System (1998-2000) were compared with figures from previous analyses for 1990-1997. RESULTS: The percentage of people with gambling experience increased, but the proportion of currently active gamblers decreased. The proportion of frequent gamblers (as an indicator of high risk behaviour) did not change. The annual average number of new outpatient treatment cases per treatment facility increased, from a low baseline of 1.5 cases, by about 20 %. All figures are still below those for former Western Germany. CONCLUSION: At least until 2000 A.D., the expected extreme increase in gambling behaviour and related treatment demand did not occur. PMID- 12548485 TI - [Public health in a European context -- comment by the Public Health Service on the promotion of health preventive measures in Germany]. PMID- 12548486 TI - What's new in minimally invasive paediatric surgery? AB - There has been substantial progress in minimally invasive techniques in children in recent years. Numerous investigators have shown that laparoscopic and thoracoscopic paediatric surgery can be performed safely. It has been postulated that minimally invasive paediatric surgery is associated with low morbidity, a shorter hospital stay, lower costs and clinical results similar to those achieved by open surgery. The present article reviews information from the past 2 years on the pathophysiological effects, feasibility, standards, new techniques, the importance of the method in malignant diseases and robotics. However, most of these reports still focus on safety and feasibility. It would be preferable if the paediatric surgical society itself would discriminate between useful and useless or harmful techniques. Therefore, further systematic research on the advantages and limits of minimally invasive surgery in children is mandatory. PMID- 12548487 TI - PHACE syndrome: new views on diagnostic criteria. AB - The association of large facial hemangiomas with posterior fossa malformations and vascular anomalies has been termed the PHACE syndrome. It is characterized by the association of posterior fossa malformations, hemangiomas, arterial anomalies, coarctation of the aorta and other cardiac defects, and eye abnormalities. Since most articles focus on isolated case reports, an extended retrospective literature review of all reports of large hemangiomas with associated abnormalities of the central nervous system and other malformations was performed to examine the clinical features, and other not as yet reported associated anomalies. Reports were found on 59 patients with PHACE syndrome, to which we added ten cases of our own. The Dandy-Walker syndrome is the most common CNS abnormality reported in association with PHACE syndrome and was seen in 48 (81 %) patients. Arterial malformations were found in 13 (22 %) cases; only 11 patients (19 %) had structural arterial abnormalities without associated Dandy Walker complex. As published, about one third of patients (31 %) had further ophthalmologic abnormalities, and cardiac anomalies, including coarctation of the aorta. Subglottic hemangiomas were seen in 4 (7 %) patients and ventral developmental defects also in 3 cases. In seven of 59 patients (12 %) with PHACE syndrome, intracranial hemangiomas were present. This study demonstrates that among other CNS abnormalities, special attention should be given to intracranial hemangiomas which seems to be a peculiar phenotype of PHACE syndrome. We therefore suggest that a sixth criterion should be added to the five minimal inclusion criteria for PHACE syndrome. The inclusion criteria would then be: arterial abnormalities or/and intracranial hemangiomas. On the basis of our experience with our patients and with those previously reported, we stress the importance of using contrast-enhanced imaging to detect intracranial lesions. PMID- 12548488 TI - Paediatric gastric teratoma. AB - AIM: This paper reviews our experience with patients presenting with gastric teratoma treated over an eighteen-year period. This rare paediatric tumour has an excellent prognosis after curative resection. METHODS: There were seven patients, all males. Investigations included a haematological profile, plain radiography and ultrasound of the abdomen. All patients were operated and the specimens subjected to histopathological examination. RESULTS: The age at presentation ranged from 1 month to 7 years. Two patients presented with melaena and the rest with progressively increasing abdominal mass. Our oldest patient had a massive tumour occupying nearly the whole abdomen. None of the children had metastases and none of the lesions had invaded adjacent structures. The tumours could be completely resected and all patients had an uneventful recovery. Histology of the tumours showed mature elements arising from the three germ layers in 6 children, one child had an immature teratoma. The average follow-up was 3 years (range 2 - 6 years). None of the patients has had a recurrence or any other problem. CONCLUSIONS: Gastric teratoma is a rare tumour but amenable to curative resection with an excellent prognosis. PMID- 12548489 TI - Ultrasonographic follow-up of infantile hypertrophic pyloric stenosis after pyloromyotomy: a controlled prospective study. AB - Although the ultrasonographic assessment of hypertrophic pyloric stenosis (IHPS) has become the main radiological method for this disease, our knowledge about postoperative ultrasonographic follow-up is very limited. To evaluate the ultrasonographic outcome of the pylorus, we performed ultrasonographic measurements of the pylorus in 22 children with IHPS, before and after the operation in a prospective trial. The sonograms following surgery were undertaken at the end of the 1st postoperative week and of the 1st, 3rd and 6th month postoperatively. Three main ultrasonographic parameters were used: pyloric muscle thickness (MT), pyloric diameter (PD) and pyloric length (PL). The values of the pylorus, especially the "MT" measurements, began to decrease almost by the end of the first postoperative week, and were normal by the 3rd month in the majority of cases. However, the averages of the PL and PD values never returned to normal throughout the 6 months follow-up. There was a highly significant difference between the average values of pyloric muscle thickness at admission and at the 6th month after surgery (p < 0.001). Within one month after surgery, the ultrasonographic parameters returned to the levels of those in control infants. However, except for MT, they did not return to their normal ranges. Therefore, MT is the most useful parameter for ultrasonographic postoperative evaluation if the normal ultrasonographic values, as given in the literature, are used. PMID- 12548490 TI - Transumbilical laparoscopic-assisted appendectomy (TULAA): a safe and useful alternative for uncomplicated appendicitis. AB - AIM: To compare transumbilical laparoscopic-assisted appendectomy (TULAA) and open appendectomy (OA) in cases of acute uncomplicated appendicitis. METHODS: 58 TULAA and 65 OA in patients with a preoperative diagnosis of acute uncomplicated appendicitis are compared. Pneumoperitoneum was obtained with a transumbilical 10 mm trocar (telescope access) and a 5 mm operative channel introduced in the left iliac fossa. Appendectomy was performed outside the abdomen, after the exteriorisation of the appendix through the transumbilical incision. RESULTS: In the TULAA group, operative time and hospital stay were reduced. Conversion was necessary in one case (1.7 %), and in one case (1.7 %), an additional 5 mm operative channel was introduced. Neither intra- or postoperative complications were found in the TULAA group, with excellent cosmetic results. In the OA group we had a wound infection (1.5 %) and in 8 cases (12.3 %) an enlargement of the incision was necessary. CONCLUSIONS: TULAA is the best approach in uncomplicated appendicitis. It is less invasive and traumatic, permits a complete evaluation of the peritoneal cavity with superior cosmetic results, especially in obese patients and in cases of ectopic appendicitis. PMID- 12548491 TI - Air reduction of intussusception. AB - AIM: To show that intussusception in children is reducible in most cases, using air. METHODS: Initially we used a barium enema to treat paediatric intussusception. Now we use an air enema. We consider the latter reduction technique to be the method of choice in the treatment of intussusception in childhood. RESULTS: Over a period of 21 years, we have treated 151 patients presenting with intussusception. In a group of 130 children treated with barium enema we managed to reduce the intussusception in 110 cases (84.6 %). We used air pressure to treat a second group of 21 children; the success rate in this group was 100 %. CONCLUSIONS: In order to avoid a surgical operation this technique must become more generally known. We are of the opinion that if the method is applied correctly, it is almost always possible to correct intussusception using air. PMID- 12548492 TI - Mouse-isolated plexus differentiates neural crest precursors into enteric neuroblasts. AB - Aim of this study was to investigate, for the first time, whether isolated newborn mouse enteric plexus could induce in vitro differentiation of the vagal neural crest-derived cells into enteric neuroblasts. Fragments of the myenteric plexus were isolated from the small intestine of 6-day-old Swiss mice and were collected and stored in DMEM-F12 medium, then cultured on polymerized human fibronectin layer. The vagal portion of the neural tube, isolated from a 9.5-day old Swiss mouse embryo, was put in the same chamber slides where the isolated myenteric plexus had been cultured for 3 days. The vagal neural crest-derived cells migrated onto the polymerized human fibronectin layer and formed a crown of cells around the neural tube. After 6 days, the cultures were stopped and studied immunohistochemically for anti-NF160 KD, anti-TH, and RetR5 antibodies to analyse the differentiation stage of the cultured cells. Analysis of results included the comparison of two culture groups: Group 1, used as control, in which vagal neural crest-derived cells were put in DMEM-F12, supplemented only with 10 % of FCS; Group 2, in which vagal neural crest-derived cells were put in the same medium as Group 1, with the addition of myenteric plexus fragments isolated from newborn mice to form the co-culture. The following results were obtained: in Group 1 the neural tubes originated a cell population strongly positive for anti-NF160 and anti-TH Ab, but negative for RetR5 Ab. This positivity was found both in the cells adjacent to the neural tube and in those migrating from it distally. The Group 2 originated cells, which after migration were positive for anti-NF160 and for anti-TH antibodies. In addition, in this culture group, the cells which migrated from the neural tube were positive for anti-RetR5 antibody. The co culture used in this study induces the differentiation of vagal stem cells into enteric neuroblasts, cells TH+ and RetR5+. These cells, after reaching the embryonic intestine, migrate to colonize the hindgut and form the ENS. Therefore this biotechnology seems a good method to obtain in vitro enteric precursors of ENS. PMID- 12548493 TI - Suitability of selected markers for identification of elements of the Intestinal Nervous System (INS). AB - The possibility of identifying and characterising elements of the enteric nervous system based on their contents of cathepsin D, chromogranin A, neuronal specific enolase and S-100 protein was studied in colorectal specimens (operative full thickness, seromuscular and mucosomuscular biopsies) obtained from 15 children, aged 2 days to 10 years. Nine patients suffered from Hirschsprung's disease, and two from chronic constipation. Four neonates with imperforate anus or meconium ileus composed the control group. All markers were identified immunohistochemically by antibodies against human antigens with appropriate detection methods. Chromogranin A staining was not always adequate to identify all neuronal cell bodies and other nervous elements. However, it proved superior to the other methods in the depiction of neuroendocrine cells in the intestinal mucosa. Cathepsin D antibodies stained normal and abnormal neural cells with different intensity; nerve fibres were not stained. This marker did not allow an unequivocal differentiation of ganglion cells from macrophages within the submucosa; the latter exhibited exceptionally strong marking and in some cases represented the predominant elements in this area. Neuronal specific enolase was distinctly expressed in nerve cells and fibres of the intestinal wall. Atrophic and hypoplastic features could be identified, suggesting that this method may give some insight into functional aspects. Continuous connections between ganglions were also observed. S-100 protein antibodies resulted in a negative image of unstained ganglion cells surrounded by extensively marked Schwann cells and neural fibres. With respect to clinical application, all these markers may provide supplementary information for the differential diagnosis of intestinal motility disorders. PMID- 12548494 TI - The impact of surgical radicality on outcome in childhood neuroblastoma. AB - AIM: Improvement of treatment results for neuroblastoma (NB) has been achieved during recent years, especially by intensifying therapy for advanced NB. Surgery, however, has not contributed very much to this progress and there is still controversy regarding the best approach for high-risk NB. We therefore attempted to find criteria for a differentiated strategy for tumour resection in NB. METHODS: We retrospectively analysed the data of 2251 NB patients treated in the German Cooperative NB Studies NB79 - NB97 (1979 - 1999) including patients' age, tumour stage, MYCN oncogene status, surgical intervention, completeness of resection, surgical complications and outcome. RESULTS: 1148 patients had a localised NB (stage 1 - 3), 878 had stage 4, and 225 had stage 4 S disease. 2112 patients underwent surgery. Tumour resection as the final result of primary or delayed operation was complete in 1403 (66.4 %), incomplete in 449 (21.3 %), and only a biopsy was carried out in 260 (12.3 %) cases. Complete resection was performed most often in localised NB (73.5 %), less often in stage 4 NB (59.2 %) and in stage 4 S (54.5 %). Clinically relevant complications occurred in 19.2 % of all operations independent of the time of resection during treatment, patients' age, tumour stage, tumour site and MYCN status. For 1787 patients from the studies NB79 - NB90 the probability of 5-year event-free survival (EFS) could be analysed and correlated with surgical radicality. In localised NB of patients aged > 1 year there was a significant difference in 5-year EFS between complete, incomplete, and no substantial resection ( P < 0.0001), while this was not the case in infants < 1 year. In the early study period (NB79 - NB85) patients with stage 4 NB had a significantly better 5-year EFS after complete excision of the primary tumour. In contrast, this difference was not confirmed during the later period (NB90) with intensified drug therapy. Timing of surgery did not influence prognosis. For infants with stage 4 S NB there was no difference in outcome after complete or incomplete resection of the primary tumour. MYCN status did not alter the significant relevance of tumour resection for outcome in localised NB. In contrast to MYCN non-amplified stage 4 NB, however, stage 4 NB with MYCN amplification had a significantly better outcome if the primary tumour could be completely or incompletely resected, compared to a biopsy only. CONCLUSIONS: Radical tumour resections with the risk of surgical complications are only justified in children > 1 year with a localised NB. In stage 4 NB, the primary tumour may be resected during intensive multimodal treatment without risky complications. Complete or incomplete resection of the primary tumour improves prognosis only in MYCN-amplified stage 4 NB. Stage 4 S NB with non-amplified MYCN are subject to spontaneous regression; a resection after chemotherapy may be indicated in cases of non-sufficient regression or growth of the primary tumour. PMID- 12548495 TI - Biomechanical evaluation of the value of osteosynthesis in supracondylar fracture of the humerus using Kirschner pins in children. AB - The subject under discussion here is one of the most frequent and most typical fractures in children. The use of Kirschner pins is not in question, but their number and position is. We present here a biomechanical analysis of how the train force affects the fissure in supracondylar fractures of the humerus with different forms of osteosynthesis using Kirschner pins, and how stable the osteosynthesis is. The experiments were carried out at the Laboratory of Biomechanics in the Institute of Anatomy of the Medical Faculty of Zagreb. PMID- 12548496 TI - Five-year follow-up of intermittent distracting rod correction in congenital scoliosis. AB - Severe progressive curve deformities in children's congenital scoliosis often require an early operative intervention. As spinal fusion interferes with longitudinal growth, a "growth enabling" operative device may represent an alternative method to delay a progressive curve deformity until an eventual operative intervention can be carried out. We report on the five-year follow-up of a seven-year-old girl who presented at our clinic with severe congenital scoliosis of 62 degrees with a unilateral unsegmented bar between T7 and T12 and an anomaly of segmentation between L2 and L4. An intermittent distracting rod was implanted with transpedicular fixation from T3/T5 to L3/L4. The correction was obtained by distraction of the mobile segments and also by hemiepiphyseodesis between L2 and L4. The rod's longitudinal extension could be distracted using a special thread. Three successive operations to obtain a minimal access to the thread were necessary to achieve a correction to an angle of 30 degrees with a total rod distraction of 4.5 cm. PMID- 12548497 TI - Tracheobronchial rupture due to blunt trauma in children: report of two cases. AB - Tracheobronchial tree injuries occur in a small number of patients after blunt chest trauma, and their occurrence is uncommon in the pediatric trauma population. The authors report two male children, one with a tracheal rupture, and the other with disruption of the main right bronchus. Mediastinal and subcutaneous emphysema resulting in airway obstruction were noted in Case 1 and soft-tissue emphysema, pneumomediastinum and tension pneumothorax were evident in Case 2 at the time of presentation. In the child with bronchial disruption, a major airway injury was suspected early on, because of a massive air leak despite two properly placed chest tubes. The definitive diagnosis was established bronchoscopically, and thoracotomy and primary repair were performed. The child with rupture of the posterior tracheal wall was diagnosed at an early stage by bronchoscopy and he was successfully managed without surgery. PMID- 12548498 TI - Simultaneous correction of duodenal atresia due to annular pancreas and malrotation by laparoscopy. AB - We report the case of a 3450 g newborn girl with prenatally diagnosed duodenal obstruction. At operation, duodenal atresia due to annular pancreas and intestinal, partially volvulated malrotation became apparent. Surgical correction was completely laparoscopic with 3-mm instruments. The operation consisted of reduction of the volvulated bowel loops, division of obstructing bands, and creation of a side-to-side duodenoduodenostomy. The technique, described in detail, proved to be feasible. No postoperative complication occurred and the girl is doing well 4 months postoperatively. PMID- 12548499 TI - Intra-abdominal extra lobar pulmonary sequestration communicating with gastric duplication -- a case report. AB - Gastric duplications with associated pulmonary sequestration are very infrequent abnormalities. Although often asymptomatic, they should be considered in the differential diagnosis of intra-abdominal or retro-peritoneal masses. The authors report the case of a ten-year-old boy who had an occasional finding of poorly symptomatic intra-abdominal mass, recognised at intervention as a gastric duplication with associated extralobar pulmonary sequestration. The authors describe the surgical treatment of this case and briefly discuss the current views on the embryological pathogenesis of such malformations. PMID- 12548500 TI - Intestinal pseudoobstruction as a paraneoplastic syndrome in ganglioneuroblastoma. AB - Intestinal pseudoobstruction may be part of a paraneoplastic syndrome. We report a teenage girl with ganglioneuroblastoma who presented with severe constipation. The intestinal pseudoobstruction was presumed to be due to inflammation of the myenteric plexus with destruction of the ganglion cells caused by antineuronal nuclear antibodies (ANNA or Anti-Hu). PMID- 12548501 TI - Failure to find DUP25 in patients with anxiety disorders, in control individuals, or in previously reported positive control cell lines. AB - Investigation of the co-occurrence of panic and phobic disorders with joint laxity led to the identification of various forms of interstitial duplications involving human chromosome 15q24-q26 (named "DUP25") in a Spanish population. DUP25 was observed in 68 of 70 (97%) patients assigned the diagnosis panic disorder/agoraphobia. DUP25 was also found in 14 of 189 (7%) control individuals. In the present study, we replicated the experimental conditions described by Gratacos and colleagues in which fluorescence in situ hybridization was used to examine metaphase chromosomes of patients with panic disorder/social phobia and of control individuals from a southern region of the United Kingdom, the primary aim being to determine the prevalence of this chromosomal rearrangement in a geographically and ethnically distinct population. DUP25 was not observed in any of our 16 patients or 40 control samples or in three previously reported DUP25 positive control (Centre d'Etude du Polymorphisme Humain) cell lines, indicating a highly significant difference in the frequency of DUP25 between the study by Gratacos and colleagues and the present investigation. PMID- 12548502 TI - Effects of tacrolimus on ischemia-reperfusion injury. AB - In addition to efficacious immunosuppression for the benefit of organ transplantation, tacrolimus has diverse actions that result in amelioration of ischemia-reperfusion injury. Knowledge is accumulating rapidly on the mechanisms through which tacrolimus exerts these cytoprotective effects, including alterations in microcirculation, free radical metabolism, calcium-activated pathways, inflammatory cascades, mitochondrial stability, apoptosis, stress response proteins, and tissue recovery. Within the nucleus, actions mediating the effects of tacrolimus appear to be dominantly influenced by interactions with the transcription factor, nuclear factor-kappaB. Because tacrolimus is a cornerstone agent in immunosuppression regimens throughout the world and knowledge of its cellular mechanisms is evolving, it is important to update the clinical literature with this information. We reviewed the published literature with intent to portray the interactions of tacrolimus in the intricate cellular mechanisms initiated by ischemia and reperfusion. PMID- 12548503 TI - A correlation between the pretransplantation MELD score and mortality in the first two years after liver transplantation. AB - The Model for End-Stage Liver Disease (MELD) score is now the criteria for allocation in liver transplantation for patients with chronic disease. Although the score has been effective in the prediction of mortality in patients awaiting liver transplantation, its abilities to predict posttransplantation outcome need study. The aim of this study is to compare outcome in the first 2 years after liver transplantation according to the pretransplantation MELD score. The study includes 669 consecutive patients who underwent primary liver transplantation between December 1993 and October 1999 in a single transplant center. Patients who died of malignancy were excluded from the series. Pretransplantation MELD score was calculated using the United Network for Organ Sharing formula. Patients were stratified according to MELD score less than 15, 15 to 24, and 25 and higher. Posttransplantation survival at 3, 6, 12, 18, and 24 months was significantly lower in the groups with a higher MELD score. The difference was significant for hepatitis C and noncholestatic liver diseases, but not cholestatic diseases. In patients with a MELD score between 15 and 24, survival was significantly greater with cholestatic diseases and lower in patients with hepatitis C. In our study, pretransplantation MELD score correlates with survival in the first 2 years after transplantation. There is a survival advantage for patients with cholestatic diseases compared with those with hepatitis C. These findings suggest the need to readjust MELD score-based allocation decisions to consider patient outcome. PMID- 12548504 TI - Pretransplantation disease severity and posttransplantation outcome. PMID- 12548505 TI - Sirolimus monotherapy in nephrotoxicity due to calcineurin inhibitors in liver transplant recipients. AB - Sirolimus, being nonnephrotoxic, is a viable alternative in patients who develop renal insufficiency caused by calcineurin inhibitors (CIs). The aim of this study is to determine whether there is improvement in renal function in liver transplant recipients after switching to sirolimus-based immunosuppression. In this retrospective review, patients who were more than 3 years posttransplantation were selected. Patients who had proteinuria (protein > 300 mg/24 hr), those administered any other nephrotoxic agents, and those with a creatinine clearance (CCr) less than 20 mL/min were excluded. Renal insufficiency was defined as mild (CCr > 70 mL/min), moderate (CCr, 40 to 70 mL/min), or severe (CCr, 20 to 40 mL/min). In the 16 patients studied; there was significant improvement in serum blood urea nitrogen (36 mg/dL; range, 19 to 53 mg/dL; to 25 mg/dL; range, 10 to 37 mg/dL; P =.002) and serum creatinine levels (median, 1.95 mg/dL; range, 1.3 to 2.8 mg/dL; to 1.5 mg/dL; range, 1.0 to 2.4 mg/dL; P =.001) 6 months after switching to sirolimus therapy. There also was a trend in improvement in CCr from 43 mL min (range, 24 to 68 mL/min) to 49 mL/min (range, 22 to 152 mL/min). Among 9 patients with moderate renal insufficiency, 2 patients improved to mild renal insufficiency, 4 patients remained unchanged, and 3 patients deteriorated to severe renal insufficiency. Among 7 patients with severe renal insufficiency, 1 patient improved to mild renal insufficiency, 4 patients improved to moderate renal insufficiency, and 2 patients remained unchanged. No patient developed cellular rejection or other graft-related complications. In liver transplant recipients with chronic renal insufficiency, conversion to sirolimus-based immunosuppression allows complete withdrawal of CIs, leading to some improvement in renal function. PMID- 12548506 TI - Toward better outcomes with tacrolimus therapy: population pharmacokinetics and individualized dosage prediction in adult liver transplantation. AB - Patient outcomes in transplantation would improve if dosing of immunosuppressive agents was individualized. The aim of this study is to develop a population pharmacokinetic model of tacrolimus in adult liver transplant recipients and test this model in individualizing therapy. Population analysis was performed on data from 68 patients. Estimates were sought for apparent clearance (CL/F) and apparent volume of distribution (V/F) using the nonlinear mixed effects model program (NONMEM). Factors screened for influence on these parameters were weight, age, sex, transplant type, biliary reconstructive procedure, postoperative day, days of therapy, liver function test results, creatinine clearance, hematocrit, corticosteroid dose, and interacting drugs. The predictive performance of the developed model was evaluated through Bayesian forecasting in an independent cohort of 36 patients. No linear correlation existed between tacrolimus dosage and trough concentration (r(2) = 0.005). Mean individual Bayesian estimates for CL/F and V/F were 26.5 +/- 8.2 (SD) L/hr and 399 +/- 185 L, respectively. CL/F was greater in patients with normal liver function. V/F increased with patient weight. CL/F decreased with increasing hematocrit. Based on the derived model, a 70-kg patient with an aspartate aminotransferase (AST) level less than 70 U/L would require a tacrolimus dose of 4.7 mg twice daily to achieve a steady-state trough concentration of 10 ng/mL. A 50-kg patient with an AST level greater than 70 U/L would require a dose of 2.6 mg. Marked interindividual variability (43% to 93%) and residual random error (3.3 ng/mL) were observed. Predictions made using the final model were reasonably nonbiased (0.56 ng/mL), but imprecise (4.8 ng/mL). Pharmacokinetic information obtained will assist in tacrolimus dosing; however, further investigation into reasons for the pharmacokinetic variability of tacrolimus is required. PMID- 12548507 TI - Recombinant activated factor VII for coagulopathy in fulminant hepatic failure compared with conventional therapy. AB - Severe coagulopathy in fulminant hepatic failure (FHF) is difficult to correct by conventional means. Recombinant activated factor VII (rFVIIa) is an antihemophilic factor that has shown promise in treating coagulopathy in liver disease. Our aim is to review our experience with rFVIIa in treating the coagulopathy of FHF and compare these results with those of conventional therapy. Fifteen patients with FHF who met King's College criteria for orthotopic liver transplantation were studied. All were ascertained from our liver disease registry. Eight consecutive patients were administered fresh frozen plasma (FFP) alone, whereas seven consecutive patients were administered FFP and rFVIIa (40 microg/kg intravenous bolus). The two groups, with similar demographic characteristics, were compared in terms of measured parameters of coagulopathy (prothrombin time and international normalized ratio), amount of plasma infused, development of anasarca, ability to undergo intracranial pressure (ICP) transducer placement, bleeding complications, ability to undergo transplantation, and survival. All patients administered rFVIIa (after a single dose) versus none administered FFP alone had temporary (2- to 6-hour) correction of coagulopathy (P <.0002). All patients administered rFVIIa versus 38% administered FFP alone were able to have an ICP transducer placed (P =.03). The rFVIIa group had less anasarca (P =.04). An equal number of patients underwent transplantation from each group, but overall survival was slightly better in the rFVIIa group (P =.04). Five of seven patients in the rFVIIa group were administered one or more subsequent doses of rFVIIa after placement of the ICP monitor (two patients, for additional procedures; three patients, prophylactically in the first 24 hours after ICP transducer placement) at the discretion of the attending physicians. We conclude that rFVIIa is effective in transiently correcting laboratory parameters of coagulopathy in patients with FHF. It facilitates the performance of invasive procedures and is associated with less frequent anasarca compared with conventional therapy. Our preliminary experience supports the need for further studies to define the optimal dosing, safety, and efficacy of rFVIIa in patients with FHF. PMID- 12548508 TI - Tacrolimus as a liver flush solution to ameliorate the effects of ischemia/reperfusion injury following liver transplantation. AB - The goal of this report is to evaluate in a prospective randomized fashion the effect of flushing hepatic allografts with tacrolimus before transplantation. A prospective, double-blinded, randomized trial was performed. Twenty patients receiving orthotopic liver transplants from October 2000 to October 2001 were randomized into two groups. Group 1 (active) was administered tacrolimus, 20 ng/mL, plus Plasma-lyte A (Baxter Healthcare Corp, Deerfield, IL) liver flush solution; and group 2 (placebo) was administered only Plasma-lyte A. Ischemia/reperfusion injury was assessed in both groups after transplantation by means of serum laboratory values to assess hepatocellular damage, synthetic function, and ion transport capacity. Peak values were recorded for each parameter, and their distributions were compared. There were no statistically significant differences between groups for age, sex, total ischemia time, or cause of liver disease. Global multivariate comparison of peak changes in all measures of liver function indicated liver injury was significantly lower with tacrolimus treatment than placebo (P =.01). The sample median for group 1 was less than for group 2 in all parameters measured. Individual statistical comparison showed that peak changes from baseline aspartate aminotransferase and activated partial thromboplastin time values were significantly improved (P 10%) and provided gains in life expectancy of 4.8-6.1 months with an acceptable cost (US dollars 40,000/ year of life gained) for waiting lists > or = 1 year whereas it was not cost effective (US dollars 74,000/life of year gained) for shorter waiting times or high dropout rate scenarios. Percutaneous treatment increased life expectancy by 5.2-6.7 months with a marginal cost of approximately US dollars 20,000/year of life gained in all cases, remaining cost effective for all waiting times. CONCLUSIONS: Adjuvant therapies for HCC while waiting for liver transplantation provide moderate gains in life expectancy and are cost effective for waiting lists of one year or more. For shorter waiting times, only percutaneous treatment confers a relevant survival advantage. PMID- 12548517 TI - Tapering off steroids three months after liver transplantation is not detrimental for hepatitis C virus disease recurrence. PMID- 12548518 TI - Is there a necessity to implement ex vivo isolated, extracorporeal perfusion techniques of hepatic grafts in liver transplant medicine? PMID- 12548519 TI - What causes increased muscle stiffness in cerebral palsy? PMID- 12548520 TI - Stem cell route to neuromuscular therapies. AB - As applied to skeletal muscle, stem cell therapy is a reincarnation of myoblast transfer therapy that has resulted from recent advances in the cell biology of skeletal muscle. Both strategies envisage the reconstruction of damaged muscle from its precursors, but stem cell therapy employs precursors that are earlier in the developmental hierarchy. It is founded on demonstrations of apparently multipotential cells in a wide variety of tissues that can assume, among others, a myogenic phenotype. The main demonstrated advantage of such cells is that they are capable of colonizing many tissues, including skeletal and cardiac muscle via the blood vascular system, thereby providing the potential for a body-wide distribution of myogenic progenitors. From a practical viewpoint, the chief disadvantage is that such colonization has been many orders of magnitude too inefficient to be useful. Proposals for overcoming this drawback are the subject of much speculation but, so far, relatively little experimentation. This review attempts to give some perspective to the status of the stem cell as a therapeutic instrument for neuromuscular disease and to identify issues that need to be addressed for application of this technology. PMID- 12548521 TI - Iatrogenic and toxic myopathies. AB - There has been increasing awareness of the adverse effects of therapeutic agents and exogenous toxins on the structure and function of muscle. The resulting clinical syndrome varies from one characterized by muscle pain to profound myalgia, paralysis, and myoglobinuria. Because toxic myopathies are potentially reversible, their prompt recognition may reduce their damaging effects or prevent a fatal outcome. Interest in the toxic myopathies, however, derives not only from their clinical importance but also from the fact that they serve as useful experimental models in muscle research. Morphological and biochemical studies have increased our understanding of the basic cellular mechanisms of myotoxicity. Toxins may produce, for instance, necrotizing, lysosomal-related, inflammatory, anti-microtubular, mitochondrial, hypokalemia-related, or protein synthesis related muscle damage. PMID- 12548523 TI - Severe prognosis in a large family with hypokalemic periodic paralysis. AB - Hypokalemic periodic paralysis (HypoPP) is a channel disorder caused primarily by mutations in the human skeletal muscle alpha1 subunit (CACNA1S) of the dihydropyridine-sensitive calcium channel. Molecular, clinical, and biochemical studies were aimed at establishing genotype/phenotype correlations in a large Italian family affected by a severe form of HypoPP. Whereas patients with HypoPP usually show a normal life span, in this family three male patients died young, one of them from anesthetic complications resembling malignant hyperthermia. Our patients carried the c1583G>A genetic lesion (R528H), which has been associated with a mild phenotype and with incomplete penetrance in women. Surprisingly, the R528H amino acid substitution in the family presented here correlated with an unfavorable prognosis in both male and female patients. We conclude that genetic characterization is an important requirement to alert physicians about the management of similar patients, especially when anesthesia is considered. PMID- 12548522 TI - Spastic muscle cells are shorter and stiffer than normal cells. AB - The mechanical properties of isolated single muscle fiber segments were measured in muscle cells obtained from patients undergoing surgery for correction of flexion contractures secondary to static perinatal encephalopathy (cerebral palsy). "Normal" muscle cells from patients with intact neuromuscular function were also mechanically tested. Fiber segments taken from subjects with spasticity developed passive tension at significantly shorter sarcomere lengths (1.84 +/- 0.05 microm, n = 15) than fibers taken from normal subjects (2.20 +/- 0.04 microm, n = 35). Elastic modulus of the stress-strain relationship in fibers from patients with spasticity (55.00 +/- 6.61 kPa) was almost double that measured in normal fibers (28.25 +/- 3.31 kPa). The fact that these muscle cells from patients with spasticity have a shorter resting sarcomere length and increased modulus compared with normal muscle cells suggests dramatic remodeling of intracellular or extracellular muscle structural components such as titin and collagen. Such changes in muscles of patients with spasticity may have implications for therapy. PMID- 12548524 TI - Electrophysiologic changes during exercise testing in patients with chronic obstructive pulmonary disease. AB - To determine whether skeletal muscle is involved in the exercise limitation of chronic obstructive pulmonary disease (COPD), we investigated electrical adaptations in muscle during incremental cycling exercise testing. Changes in quadriceps activity were compared using surface electromyography (SEMG) and motor point stimulation in ten COPD patients and ten healthy subjects. Patients showed significantly lower exercise capacity, and M-wave duration was increased from exercise onset (P < 0.05) with a parallel decrease in amplitude (P < 0.05). The SEMG power spectrum median frequency was always higher (P < 0.04) in patients and its decline was earlier (P < 0.01). The ratio of the root mean square of the SEMG to oxygen uptake was decreased (P < 0.001) during exercise in patients, although it remained constant in controls. Electromyographic parameters were significantly more involved in the exercise limitation than ventilatory factors. Thus, modified electrical activity in muscle appeared in COPD patients from exercise onset, indicating that skeletal muscle function is clearly implicated in the exercise intolerance of these patients. PMID- 12548525 TI - Choline acetyltransferase mutations in myasthenic syndrome due to deficient acetylcholine resynthesis. AB - The myasthenic syndrome due to abnormal acetylcholine resynthesis is characterized by early onset, recessive inheritance, and recurrent episodes of potentially fatal apnea. Mutations in the gene encoding choline acetyltransferase (CHAT) have been found to account for this condition. We have identified five patients from three independent families with features of this disease including, in four patients, a paradoxical worsening of symptoms with cold temperatures. Electrodiagnostic studies demonstrated impaired neuromuscular transmission in all patients. In vitro microelectrode studies performed in the anconeus muscle biopsies of two patients showed moderate reduction of quantal release. Electron microscopy of the neuromuscular junction was normal in both patients. Each patient had two heterozygous CHAT mutations including L210P and P211A (family 1), V194L and V506L (family 2), and R548stop and S694C (family 3). Three of these mutations have previously been reported and suggest that, in this syndrome, some molecular defects may be more prevalent than others. PMID- 12548526 TI - Sensory neuropathy with monoclonal IgM binding to a trisulfated heparin disaccharide. AB - We studied clinical and serological features of five patients with polyneuropathy and serum immunoglobulin M (IgM) binding to the trisulfated disaccharide IdoA2S GlcNS-6S (TS-HDS), the most abundant disaccharide component of heparin oligosaccharides. The patients all had painful, predominantly sensory polyneuropathies. Sensory loss was distal and panmodal. Electrophysiological and pathological studies were consistent with axonal loss, especially of unmyelinated axons. Immunohistochemistry showed IgM and kappa light chains deposited around the rim of intermediate-sized veins in the perimysium and epineurium. Serum IgM binding to TS-HDS was selective, present in high titer (>12,000), and limited to kappa light chains. We conclude that TS-HDS is a newly identified target carbohydrate antigen of some IgM M-proteins. Monoclonal IgM binding to TS-HDS is associated with a painful, predominantly sensory, polyneuropathy syndrome with axonal loss and deposition of IgM in veins. The role of IgM binding to TS-HDS in the pathogenesis of the neuropathy remains to be determined. PMID- 12548527 TI - Size of motor unit potential sample. AB - In quantitative electromyography (EMG), a sample size of 20 motor unit potentials (MUPs) is standard. The effect of increase in the number of MUPs above 20 is not known, although advanced MUP analysis techniques make such samples practical. In the present study, using multi-MUP analysis, pools of 3,720 neuropathic and 2,526 control MUPs were obtained from external anal sphincter muscles. From each pool, 10,000 random samples of 5, 10, 15, 20, 30, 40, 50, and 100 MUPs were obtained by a computer. For each sample size, 95% normative limits for mean values, SDs, and "outliers," and sensitivities were calculated for eight MUP parameters. As the magnitude of MUP samples increased, normative limits narrowed and sensitivities increased (at 5: 20-30%; at 20: 30-55%; at 100: 80-100%) for all statistics of all MUP parameters. Our results demonstrated a substantial increase in sensitivity by increasing the MUP sample to more than 20. This option deserves consideration in an attempt to improve the usefulness of quantitative EMG. PMID- 12548528 TI - Individual attributes versus composite scores of nerve conduction abnormality: sensitivity, reproducibility, and concordance with impairment. AB - Composite scores may be more sensitive and reproducible than single attributes of nerve conduction for detection of peripheral neuropathy, but this requires validation in large patient cohorts. Also, the concordance of individual attributes versus composite scores with clinical measures of severity has not been tested. Here, we study these issues in prospectively studied cohorts: diabetic patients from Rochester, Minnesota (RDNS; n = 396); chronic inflammatory demyelinating polyneuropathy (CIDP) patients (n = 55); and multifocal motor neuropathy (MMN) patients (n = 18). With specificity fixed at the 97.5 percentile, we found that, in generalized polyneuropathies (diabetic and CIDP), composite scores (especially ones including conduction velocity, distal latencies, and F-waves) of individual or multiple nerves tended to be more sensitive than individual attributes. By contrast, for multiple mononeuropathies, some individual attributes or composite scores of individual nerves were more sensitive than composite scores. In diabetic polyneuropathy, composite scores tended to be more reproducible than individual attributes of nerve conduction. Highly significant correlations were found between individual attributes or composite scores and neurologic impairment in diabetic polyneuropathy and in CIDP; in general, correlation coefficients were higher for composite scores. These correlations were higher for amplitudes than for conduction velocities or distal latencies. We conclude that, with the availability of microprocessors and normative databases, electromyographers may increasingly seek to express nerve conduction abnormality also as composite scores of individual or several nerves. PMID- 12548529 TI - Evidence for strength imbalances as a significant contributor to abnormal synergies in hemiparetic subjects. AB - Abnormal synergies in the paretic shoulder and elbow of hemiparetic subjects were quantified during maximal voluntary contractions (MVCs) in 27 subjects with a history of stroke and 8 age-matched control subjects. A six-axis load cell allowed simultaneous measurement of the primary torque the subject was attempting to maximize and the secondary torques at other joint actions. For example, during MVC of shoulder flexion, shoulder flexion is the primary torque and the secondary torques are internal/external rotation, abduction/adduction, and elbow flexion/extension. In general, the stroke subjects had increased secondary torques compared to controls, resulting in abnormal joint torque coupling within the set consisting of elbow flexion, internal rotation, adduction, shoulder flexion. Unlike previous studies, abnormal secondary torques in several cases were due to strength imbalances, which occur when the strength deficit for a particular joint action is greater than the strength deficit in the opposite joint action. This hypothesis was supported by electromyographic recordings and by the finding that subjects with larger strength imbalances tended to produce larger secondary torques. Possible mechanisms and consequences for rehabilitative treatments are discussed. PMID- 12548530 TI - Early prednisone treatment in Duchenne muscular dystrophy. AB - The purpose of this long-term, open parallel-group, double-consent study of alternate-day, low-dose prednisone in 2-4-year-old patients with Duchenne muscular dystrophy (DMD) was to determine whether prednisone produces a beneficial effect when given earlier than usual. Muscle function was evaluated by timed tests, and muscle strength with a hand-held myometer. After 55 months of treatment, the five patients (mean age 8.3 years) in the prednisone group were still able to get up from the floor, whereas two of the three in the control group had lost this ability. Side effects included a decline in growth rate in the prednisone-treated patients and excessive weight gain in one control and three treated patients. Because steroids are effective in prolonging function, but not in recovering lost function, we propose that treatment be started with low-dose prednisone in DMD patients as soon as the diagnosis is definite. PMID- 12548531 TI - Endurance training alters the biophysical properties of hindlimb motoneurons in rats. AB - The purpose of the study was to determine the effect of daily endurance treadmill training (2 h/day, 30 m/min) on motoneuron biophysical properties. Electrophysiological properties of tibial motoneurons were measured in situ in anesthetized (ketamine/xylazine) control and trained rats using sharp glass microelectrodes. Motoneurons from trained rats had significantly hyperpolarized resting membrane potentials and spike trigger levels, and faster antidromic spike rise-times. "Fast" motoneurons (after-hyperpolarization half-decay time <20 ms) in trained rats also had a significantly larger mean cell capacitance than those in control rats, suggesting that they were larger, although this had no effect on indices of excitability (rheobase, cell input resistance). Motoneurons are thus targets for activity-induced adaptations, which may have clinical significance for the role of physical activity as a therapeutic modality in cases of neurological deficit. The specific adaptations noted, which reflect alterations in ionic conductances, may serve to offset decreases in membrane excitability that occur during sustained excitation. PMID- 12548532 TI - Full-length dystrophin cDNA transfer into skeletal muscle of adult mdx mice by electroporation. AB - We showed that a LacZ expression plasmid (pCAG-lacZ) injection followed by electroporation increased the expression of the LacZ gene in the skeletal muscles of adult mdx mice up to ninefold higher as compared with simple intramuscular DNA injection. When full-length mouse dystrophin plasmid (pCAG-dys) and pCAG-lacZ were co-transfected by electroporation, 56% of dystrophin-positive fibers were stained for beta-galactosidase activity suggesting most of these myofibers are not revertants but transfected ones. Our data indicate that electroporation in vivo could introduce large full-length dystrophin cDNA into skeletal muscle of adult mdx mice. PMID- 12548533 TI - Muscle biopsy in the evaluation of patients with modestly elevated creatine kinase levels. AB - The utility of muscle biopsy in patients with modest elevations of serum creatine kinase (CK) level but normal neurological examinations and nondiagnostic electrodiagnostic studies is uncertain. We performed systematic, extensive studies on muscle biopsies of 20 such patients. A definitive diagnosis was arrived at in only 1 by histochemical studies, although 4 others demonstrated minor myopathic changes. Biochemical evaluation led to a diagnosis in an additional 5. Muscle biopsy is useful for evaluating such patients, but extensive studies of the muscle are necessary. PMID- 12548534 TI - Effects of FK506 on myasthenia gravis patients with high interleukin-2 productivity in peripheral blood mononuclear cells. AB - We compared the early effects of FK506 on clinical severity, interleukin-2 (IL-2) production by phytohemagglutinin-stimulated peripheral blood mononuclear cells (PBMs), and serum levels of acetylcholine receptor antibodies between myasthenia gravis (MG) patients with elevated (>1250 pg/ml, n = 9) or normal (<1250 pg/mL, n = 10) levels of PBM IL-2 production. Reduction in clinical severity and PBM IL-2 production were significantly greater in the patients with elevated IL-2 production than those with normal PBM IL-2 production in the first month of treatment. PMID- 12548535 TI - Fatigue and abnormal neuromuscular transmission in Kennedy's disease. AB - We describe a patient with Kennedy's disease (X-linked bulbospinal neuronopathy) who experienced leg muscle fatigue with long-distance running. The patient also reported muscle twitching involving the face and extremities and long-standing muscle cramps. Aside from mild facial and tongue weakness (and fasciculations), his examination was normal, including completely preserved muscle strength in the extremities. Electrodiagnostic evaluation revealed evidence for a chronic motor axonopathy/neuronopathy and abnormal sensory nerve action potentials. In addition, repetitive nerve stimulation studies were normal, but neuromuscular jitter tested in the same muscle was markedly abnormal. The normal clinical strength and repetitive nerve stimulation studies in a muscle showing markedly increased neuromuscular jitter suggested a mechanism for this patient's symptoms of muscle fatigue, related to failure of neuromuscular transmission at a critical number of endplates during extremes of physical activity. PMID- 12548536 TI - Weaning from the ventilator using BiPAP in myasthenia gravis. PMID- 12548537 TI - Significance of uncompacted myelin lamellae in POEMS syndrome. PMID- 12548539 TI - Hox genes as synchronized temporal regulators: implications for morphological innovation. AB - In vertebrates, clusters of Hox genes express in a nested and hierarchical fashion to endow the embryo's segments with discrete identities. Later in development, members of the same gene family are employed again to pattern the limb, intestinal, and reproductive systems. A careful analysis of the morphologies of Hox mutant mice suggests that the genes provide qualitatively different cues during the specification of segments than they do during the development of more recently derived structures. In addition to the regulatory differences noted by others, the activity of Hox genes during specification of the vertebrate metameres in some recent deletion experiments is inconsistent with a role for them as strictly spatial determinants. On the contrary, the phenotypes observed are suggestive of a role for them as elements of a generic time-keeping mechanism. By contrast, the specification of more recent evolutionary structures appears to be more spatial and gene-specific. These differences in role and effect may suggest some simple mechanisms by which the Hox clusters operate, and rules by which gene networks can be diverted to create new structures over the course of evolution. Specific predictions and experiments are proposed. PMID- 12548540 TI - Origin of feathers: Feather beta (beta) keratins are expressed in discrete epidermal cell populations of embryonic scutate scales. AB - The feathers of birds develop from embryonic epidermal lineages that differentiate during outgrowth of the feather germ. Independent cell populations also form an embryonic epidermis on scutate scales, which consists of peridermal layers, a subperiderm, and an alpha stratum. Using an antiserum (anti-FbetaK) developed to react specifically with the beta (beta) keratins of feathers, we find that the feather-type beta keratins are expressed in the subperiderm cells of embryonic scutate scales, as well as the barb ridge lineages of the feather. However, unlike the subperiderm of scales, which is lost at hatching, the cells of barb ridges, in conjunction with adjacent cell populations, give rise to the structural elements of the feather. The observation that an embryonic epidermis, consisting of peridermal and subperidermal layers, also characterizes alligator scales (Thompson, 2001. J Anat 198:265-282) suggests that the epidermal populations of the scales and feathers of avian embryos are homologous with those forming the embryonic epidermis of alligators. While the embryonic epidermal populations of archosaurian scales are discarded at hatching, those of the feather germ differentiate into the periderm, sheath, barb ridges, axial plates, barbules, and marginal plates of the embryonic feather filament. We propose that the development of the embryonic feather filament provides a model for the evolution of the first protofeather. Furthermore, we hypothesize that invagination of the epidermal lineages of the feather filament, namely the barb ridges, initiated the formation of the follicle, which then allowed continuous renewal of the feather epidermal lineages, and the evolution of diverse feather forms. PMID- 12548541 TI - Protein evolution: structure-function relationships of the oncogene beta-catenin in the evolution of multicellular animals. AB - Beta-catenin functions as a cytoskeletal linker protein in cadherin-mediated adhesion and as a signal mediator in wnt-signal transduction pathways. We use a novel integrative approach, combining evolutionary, genomic, and three dimensional structural data to analyze and trace the structural and functional evolution of beta-catenin genes. This approach also enabled us to examine the effects of gene duplication on the structure and function of beta-catenin genes in Drosophila, C. elegans, and vertebrates. By sampling a large number of different taxa, we identified both ancestral and derived motifs and residues within the different regions of the beta-catenin proteins. Projecting amino acid substitutions onto the three- dimensional structure established for mouse beta catenin, we identified specific domains that exhibit loss and gain of selective constraints during beta catenin evolution. Structural changes, changes in the amino acid substitution rate, and the appearance of novel functional domains in beta-catenin can be mapped to specific branches on the metazoan tree. Together, our analyses suggest that a single, beta-catenin gene fulfilled both adhesion and signaling functions in the last common ancestor of metazoans some 700 million years ago. In addition, gene duplications facilitated the evolution of beta catenins with novel functions and allowed the evolution of multiple, single function proteins (cell adhesion or wnt-signaling) from the ancestral, dual function protein. Integrative methods such as those we have applied here, utilizing the 'natural experiments' present in animal diversity, can be employed to identify novel and shared functional motifs and residues in virtually any protein among the proteomes of model systems and humans. PMID- 12548542 TI - Positive Darwinian selection operating on the immunoglobulin heavy chain of Antarctic fishes. AB - The cooling of the Southern Ocean to the freezing point of seawater (-1.9 degrees C) over the past 25 million years played a dominant selective role in the evolution of the Antarctic fish fauna. During this period, the perciform suborder Notothenioidei, which is largely endemic to the Antarctic, diversified and developed numerous cold-adapted characters. In this report, we provide compelling evidence that the immunoglobulin heavy chain (IgH) of the notothenioid fishes has undergone adaptive selection. Two and four IgH clones were isolated, respectively, from spleen cDNA libraries prepared from the Antarctic icefish Chaenocephalus aceratus and the yellowbelly rockcod Notothenia coriiceps. The transmembrane region of the membrane form of the rockcod IgM heavy chain was located at the end of the second constant (C(H)) domain, in contrast to other teleost IgMs in which the transmembrane region is located at the end of the third constant domain. Phylogenetic analyses of C(H) regions revealed that rates of nonsynonymous nucleotide substitution were higher than rates of synonymous nucleotide substitution. Many of the nonsynonymous substitutions introduced charge changes, consistent with positive Darwinian selection acting to adapt the structure of the notothenioid immunoglobulins. The rates of nonsynonymous nucleotide substitutions were higher than the rates of synonymous nucleotide substitutions in complementarity determining regions of variable regions, suggesting that diversity at antigen binding sites is enhanced by genomic and/or somatic selection. Results of Southern blot hybridization experiments were consistent with a translocon type of IgH gene organization reminiscent of bony fishes and tetrapods. PMID- 12548543 TI - A comparison of parthenogenetic and sexual embryogenesis of the pea aphid Acyrthosiphon pisum (Hemiptera: Aphidoidea). AB - Aphids exhibit divergent modes of embryogenesis during the sexual and asexual phases of the life cycle. To explore how a single genome can give rise to these alternative developmental modes, we have initiated embryological studies of the pea aphid, Acyrthosiphon pisum. Here we present a detailed description of parthenogenetic, viviparous embryonic development in the pea aphid. We compare and contrast development of the parthenogenetic embryo with that of the embryo resulting from sexual reproduction. The primary difference between the embryos is the scale on which development occurs: early parthenogenetic development occurs in a volume approximately three orders of magnitude smaller than the sexual egg, largely because of the apparent absence of yolk in the parthenogenetic egg. This results in a drastically different duration of syncytial energid cleavage and, presumably, patterning processes in the two embryos must act at scales that differ by orders of magnitude. The eggs also develop on time scales that differ approximately by an order of magnitude and the timing of the embryonic movements, collectively called blastokinesis, have temporally shifted relative to growth of the embryo. In addition, the endosymbiotic bacteria are transferred from mother to embryo in different ways in the two embryos. Finally, the function of the serosa has diverged greatly in the two embryos: in the sexual egg the serosa deposits a thick cuticle that protects the egg, whereas the serosa of the parthenogenetic embryo is greatly reduced and its function is unclear. The pea aphid is a useful model system for examining how a single genome has evolved to allow divergent modes of development. PMID- 12548544 TI - Telomerase activity is widespread in adult somatic tissues of Xenopus. AB - Chromosome ends, or telomeres, are maintained by telomerase. Work in selected vertebrates has implied that telomerase is often repressed in differentiated cells, and telomere erosion results in senescence of cultured cells. Tissues from mature Xenopus laevis frogs were examined for telomerase enzymatic activity with the TRAP (telomere repeat amplification protocol) assay. All tissues contained active telomerase, most abundantly in testis, spleen, liver, and embryos; activity was less abundant but still readily detectable with < 100 ng of protein extract from brain and muscle tissues. Activity in somatic tissues of the diploid Xenopus tropicalis suggests this condition is not limited to the polyploid members of the genus, and that extensive differentiation-linked telomerase repression does not occur in Xenopus. PMID- 12548545 TI - HES and HERP families: multiple effectors of the Notch signaling pathway. AB - Notch signaling dictates cell fate and critically influences cell proliferation, differentiation, and apoptosis in metazoans. Multiple factors at each step ligands, receptors, signal transducers and effectors-play critical roles in executing the pleiotropic effects of Notch signaling. Ligand-binding results in proteolytic cleavage of Notch receptors to release the signal-transducing Notch intracellular domain (NICD). NICD migrates into the nucleus and associates with the nuclear proteins of the RBP-Jkappa family (also known as CSL or CBF1/Su(H)/Lag-1). RBP-Jkappa, when complexed with NICD, acts as a transcriptional activator, and the RBP-Jkappa-NICD complex activates expression of primary target genes of Notch signaling such as the HES and enhancer of split [E(spl)] families. HES/E(spl) is a basic helix-loop-helix (bHLH) type of transcriptional repressor, and suppresses expression of downstream target genes such as tissue-specific transcriptional activators. Thus, HES/E(spl) directly affects cell fate decisions as a primary Notch effector. HES/E(spl) had been the only known effector of Notch signaling until a recent discovery of a related but distinct bHLH protein family, termed HERP (HES-related repressor protein, also called Hey/Hesr/HRT/CHF/gridlock). In this review, we summarize the recent data supporting the idea of HERP being a new Notch effector, and provide an overview of the similarities and differences between HES and HERP in their biochemical properties as well as their tissue distribution. One key observation derived from identification of HERP is that HES and HERP form a heterodimer and cooperate for transcriptional repression. The identification of the HERP family as a Notch effector that cooperates with HES/E(spl) family has opened a new avenue to our understanding of the Notch signaling pathway. PMID- 12548546 TI - Role of CTGF/HCS24/ecogenin in skeletal growth control. AB - Connective tissue growth factor/hypertrophic chondrocyte-specific gene product 24 (CTGF/Hcs24) is a multifunctional growth factor for chondrocytes, osteoblasts, and vascular endothelial cells. CTGF/Hcs24 promotes the proliferation and maturation of growth cartilage cells and articular cartilage cells in culture and hypertrophy of growth cartilage cells in culture. The factor also stimulates the proliferation and differentiation of cultured osteoblastic cells. Moreover, CTGF/Hcs24 promotes the adhesion, proliferation, and migration of vascular endothelial cells, as well as induces tube formation by the cells and strong angiogenesis in vivo. Because angiogenesis is critical for the replacement of cartilage with bone at the final stage of endochondral ossification and because gene expression of CTGF/Hcs24 predominates in hypertrophic chondrocytes in the physiological state, a major physiological role for this factor should be the promotion of the entire process of endochondral ossification, with the factor acting on the above three types of cells as a paracrine factor. Thus, CTGF/Hcs24 should be called "ecogenin: endochondral ossification genetic factor." In addition to hypertrophic chondrocytes, osteoblasts activated by various stimuli including wounding also express a significantly high level of CTGF/Hcs24. These findings in conjunction with in vitro findings about osteoblasts mentioned above suggest the involvement of CTGF/Hcs24 in intramembranous ossification and bone modeling/remodeling. Because angiogenesis is also critical for intramembranous ossification and bone remodeling, CTGF/Hcs24 expressed in endothelial cells activated by various stimuli including wounding may also play important roles in direct bone formation. In conclusion, although the most important physiological role of CTGF/Hcs24 is ecogenin action, the factors also play important roles in skeletal growth and modeling/remodeling via its direct action on osteoblasts under both physiological and pathological conditions. PMID- 12548547 TI - Pathophysiological implications of stroma pattern formation in uveal melanoma. AB - Clinical outcome of cancer patients is mainly determined by the rate of metastasis and, also by primary tumor growth. Formation of extracellular matrix and interactions of neoplastic and non-neoplastic (host) cells in solid tumors have been shown to be essential for these processes. One result of such interactions is the outgrowth of new blood vessels from existing ones, angiogenesis, to provide the tumor tissue with oxygen and nutrients. It is assumed that the neovascular bed also facilitates the escape of metastatic cells from the primary lesions. In addition, recent reports suggested the existence of blood-conducting channels lined by melanoma cells (so-called "vascular channels") accompanied by depositions of extracellular matrix patterns in cutaneous and uveal melanoma. Since the presence of these matrix structures has been negatively associated with prognosis, we hypothesize that they play a role in melanoma outgrowth or metastasis. In this review, we will discuss the morphological and functional properties of the extracellular matrix patterns in that may underlie these clinical phenomena. PMID- 12548548 TI - Cancer/testis-associated genes: identification, expression profile, and putative function. AB - Cancer/testis-associated genes (CTAs) are a subgroup of tumor antigens with a restricted expression in testis and malignancies. During the last decade, many of these immunotherapy candidate genes have been discovered using various approaches. Most of these genes are localized on the X-chromosome, often as multigene families. Methylation status seems to be the main, but not the only regulator of their specific expression pattern. In testis, CTAs are exclusively present in cells of the germ cell lineage, though there is a lot of variation in the moment of expression during different stages of sperm development. Likewise, there is also a lot of heterogeneity in the expression of CTAs in melanoma samples. Clues regarding functionality of CTAs for many of these proteins point to a role in cell cycle regulation or transcriptional control. Better insights in the function of these genes may shed light on the link between spermatogenesis and tumor growth and could be of use in anti-tumor therapies. This review outlines the CTA family and focuses on their expression and putative function during male germ cell development and melanocytic tumor progression. PMID- 12548549 TI - Expression of Smad4 in the FaDu cell line partially restores TGF-beta growth inhibition but is not sufficient to regulate fibronectin expression or suppress tumorigenicity. AB - Mutations of the Smad4 gene, a member of a group of TGF-beta signal transduction components, occur in several types of cancer suggesting that its inactivation significantly affects TGF-beta responsiveness in these tumors. To further investigate the role of Smad4 with respect to TGF-beta signaling and carcinogenesis, we re-expressed the Smad4 gene in the Smad4-deficient cancer cell line FaDu by microcell-mediated chromosome transfer (MMCT) and retroviral infection to closely approximate physiological protein levels. The Smad4 expressing FaDu clones were then evaluated for TGF-beta responsiveness to assess the role of Smad4 in TGF-beta-induced growth inhibition and target gene regulation. We found that the re-expression of the Smad4 gene by either method partially restored TGF-beta responsiveness in FaDu cells with respect to both growth inhibition and expression of p21WAF1/CIP1 and p15INK4B. However, only the microcell hybrids showed growth retardation in organotypic raft culture and an enhanced ability to upregulate fibronectin. In contrast, the re-expression of Smad4 by either method failed to suppress tumorigenicity. These results suggest that in addition to a homozygous deletion of Smad4, FaDu cells contain additional defects within the TGF-beta signaling pathway, thereby limiting the extent of TGF beta responsiveness upon Smad4 re-expression and perhaps accounting for the inability to induce p15INK4B to a high level. They also demonstrate the advantages of providing a physiological extracellular environment, when assessing TGFbeta responsiveness. PMID- 12548550 TI - INI1 expression induces cell cycle arrest and markers of senescence in malignant rhabdoid tumor cells. AB - The INI1 gene, which encodes a functionally uncharacterized protein component of the hSWI/SNF chromatin remodeling complex, is often mutated or deleted in malignant rhabdoid tumor (MRT). Two isoforms of INI1, that differ by the variable inclusion of nine amino acids, potentially are produced by differential RNA splicing. To determine the effect of the two INI1 isoforms on cell growth, INI1 devoid (MRT) and INI1-expressing cell lines were transfected separately with mammalian expression vectors or transduced with adenoviruses. Transfection of the short form of INI1 into either INI1-deficient or expressing cell lines resulted in complete suppression of cell growth in colony formation assays. The longer splice variant induced moderate to severe growth suppression of MRT cells, but had a far milder effect on non-MRT cells. Transduction of MRT cells with adenoviruses expressing either isoform of INI1 led to a dramatic change in morphology, growth suppression, and cell cycle arrest. Furthermore, senescence associated proteins were up-regulated after transduction, while levels of proteins implicated in cell cycle progression were down-regulated. Adenoviral delivery of INI1 into a non-MRT cell line, however, had no demonstrable effect on any of these parameters. These results support the genetic evidence that INI1 is a tumor suppressor gene gone awry in MRT cells, and also suggest that delivery of the INI1 gene to MRT cells by adenoviruses may lead to a more effective treatment of this highly aggressive malignancy. PMID- 12548551 TI - Dissociation of focal adhesion kinase and paxillin tyrosine phosphorylation induced by bombesin and lysophosphatidic acid from epidermal growth factor receptor transactivation in Swiss 3T3 cells. AB - Tyrosine phosphorylation of the nonreceptor tyrosine kinase p125 focal adhesion kinase (FAK) and the adapter protein paxillin is rapidly increased by multiple agonists, including bombesin (BOM) and lysophosphatidic acid (LPA), through heptahelical G protein-coupled receptors (GPCRs). The pathways involved remain incompletely understood. The experiments presented here were designed to test the role of epidermal growth factor receptor (EGFR) transactivation in the rapid increase of tyrosine phosphorylation of FAK and paxillin induced by GPCR agonists. Our results show that treatment with the selective EGFR tyrosine kinase inhibitor AG 1478, at concentrations that completely blocked the increase in tyrosine phosphorylation of these proteins induced by EGF, did not affect the stimulation of tyrosine phosphorylation of either FAK or paxillin induced by multiple GPCR agonists including LPA, BOM, vasopressin, bradykinin, and endothelin. Similar results were obtained when Swiss 3T3 cells were treated with another highly specific inhibitor of the EGF receptor kinase activity, PD-158780. Collectively, our results clearly dissociate EGFR transactivation from the tyrosine phosphorylation of FAK and paxillin induced by multiple GPCR agonists. PMID- 12548552 TI - Alpha-lipoic acid induces p27Kip-dependent cell cycle arrest in non-transformed cell lines and apoptosis in tumor cell lines. AB - alpha-Lipoic acid is a naturally-occurring co-factor found in a number of multi enzyme complexes regulating metabolism. We report here that alpha-lipoic acid induces hyperacetylation of histones in vivo and has differential effects on the growth and viability of normal versus transformed cell lines. The human tumor cell lines FaDu and Jurkat, as well as a Ki-v-Ras-transformed Balb/c-3T3 murine mesenchymal cell line, all initiated apoptosis following exposure to alpha-lipoic acid. In contrast, treatment of non-transformed cell lines with alpha-lipoic acid resulted only in reversible cell cycle arrest in G0/G1. Treatment with butyrate, another short-chain fatty acid, induced a G0/G1 arrest in both transformed and non-transformed cell lines. alpha-Lipoic acid caused a post-translational elevation in the levels of the cyclin-dependent kinase inhibitor p27Kip1. Studies using p27Kip1-deficient MEF cells demonstrated that p27Kip1 was required for the alpha-lipoic acid-mediated cell cycle arrest. The mechanism of apoptosis was independent of Fas-mediated signaling, as alpha-lipoic acid-treated Jurkat cell mutants deficient in Fas or FADD retained sensitivity to apoptosis. The differential selectivity of the pro-apoptotic effects of alpha-lipoic acid for transformed cells supports its potential use in the treatment of neoplastic disorders. PMID- 12548553 TI - Chemical sympathectomy-induced changes in TH-, VIP-, and CGRP-immunoreactive fibers in the rat mandible periosteum: influence on bone resorption. AB - The expression of neurotransmitter receptors by bone cells supports the concept that the nervous system is a regulator of bone metabolism. The discrimination of the respective roles of the sensory and sympathetic nervous systems requires evidence of topographic relationships between the corresponding fibers and the cells involved in bone turnover, in vivo. In this study, the influence of the sympathetic system on bone resorption was assessed by using a synchronized model of cortical resorption along the mandible. The sympathetic system was destroyed by daily injections of guanethidine (40 mg/kg) for 25 days; a resorption wave was induced on day 21. The distribution of periosteal tyrosine-hydroxylase (TH)-, vasoactive intestinal polypeptide (VIP)-, and calcitonin gene-related peptide (CGRP)-immunoreactive (IR) fibers was studied by compartmentalizing the periosteum. Most fibers were located in the distal, non-osteogenic compartment. TH-IR fibers were located perivascularly, VIP-IR fibers were gathered at the boundary with the osteogenic compartment, and CGRP-IR fibers were scattered. Sympathectomy decreased the number of TH- and VIP-IR fibers and increased the number of CGRP-IR fibers, without changing their topography. After the injection of Fast blue, a retrograde fluorescent marker, over the periosteum, fluorescent neuronal cell bodies were found in the superior cervical ganglion (SCG). Many neurons were TH-IR and very few were VIP-IR. Sympathectomy decreased the numbers of fluorescent and TH-IR cell bodies. It also decreased the number of preosteoclasts and osteoclasts, which had a drastic effect on the cortical bone surface, as assessed by scanning electron microscopy. These data indicate that VIP-IR fibers have a strategic position close to the most peripheral and less differentiated, osteogenic cells, pointing to a functional relationship. As poorly differentiated osteogenic cells support preosteoclast differentiation, VIP IR fibers may be involved in this process, as suggested by the smaller number of preosteoclasts in sympathectomized rats. Although VIP is predominantly a parasympathetic mediator, it seemed to be conveyed by sympathetic fibers, as shown by the marked effect of guanethidine treatment. Nevertheless, these fibers did not originate from the SCG, contrary to TH-IR fibers. PMID- 12548554 TI - Role of PP2A in the regulation of p38 MAPK activation in bovine aortic endothelial cells exposed to cyclic strain. AB - We have previously reported that cyclic strain results in rapid phosphorylation of p38 mitogen activated protein kinase (MAPKs). The aim of this study was to examine the role of protein phosphatase type 2A (PP2A) in regulating p38 MAPK activation in bovine aortic endothelial cells exposed to cyclic strain. In this study, we demonstrate that the catalytic subunit of PP2A is tyrosine phosphorylated by cyclic strain, resulting in inhibition of phosphatase activity. Okadaic acid, an inhibitor of PP2A at lower concentrations increased phosphorylation of p-38. Phospho-p38 MAPK physically associated with the catalytic subunit, PP2Ac. Phospho-p38 MAPK was dephosphorylated by purified PP2Ac in cell lysates, but if pretreated with okadaic acid, phospho-p38 MAPK was maintained. Taken together, our result suggests that PP2A plays a regulatory role in p38 MAPK activation in endothelial cells exposed to cyclic strain. PMID- 12548555 TI - HPV16 E6 and E7 oncoproteins regulate Notch-1 expression and cooperate to induce transformation. AB - Notch receptor signaling has been implicated in cellular transformation. Notch-1 receptor expression is increased during the progression from cervical intraepithelial lesions (CIN) to invasive cervical carcinoma. Moreover, the main cellular localization of Notch-1 protein changes from cytoplasmic to nuclear with the transition from CIN III to microinvasive carcinoma. Since the E6 and E7 proteins encoded by human papilloma virus (HPV) are a causative agent of cervical carcinoma, this study determined whether E6 and E7 protein expression causes the observed upregulation in Notch-1 expression. Mouse and human primary cell lines were transfected with HPV16 E6 and E7 and Notch-1 expression and activity were analyzed. We show that Notch-1 expression and activity are upregulated by E6 and E7 independently. This was due to both transcriptional and post-transcriptional mechanisms. A protein involved in Notch processing, Presenilin-1 (PS-1), was also upregulated by E6 and E7. In the presence of E6 and E7, Notch-1 protein expression is localized in the cytoplasm. Downregulation of Notch-1 expression in a human cervical carcinoma cell line expressing E6/E7 caused striking inhibition of proliferation in vitro and tumorigenicity in vivo. These data suggest that E6- and E7-mediated upregulation of Notch signaling may contribute to disruption of regular cell growth in cervical cancer. PMID- 12548556 TI - Subcellular localization of Photofrin determines the death phenotype of human epidermoid carcinoma A431 cells triggered by photodynamic therapy: when plasma membranes are the main targets. AB - Photodynamic therapy (PDT) is a kind of photochemo-therapeutic treatment that exerts its effect mainly through the induction of cell death. Distinct types of cell death may be elicited by different PDT regimes. In this study, the mechanisms involved in the death of human epidermoid carcinoma A431 cells triggered by PDT with Photofrin (a clinically approved photosensitizer) were characterized. Photofrin distributes dynamically in A431 cells; the plasma membranes and Golgi complex are the main target sites of Photofrin after a brief (3 h) and prolonged (24 h) incubation, respectively. Cells with differentially localized Photofrin displayed distinct death phenotypes in response to PDT. The effects of PDT on cells with plasma membrane-localized Photofrin were further studied in details. Cells stopped proliferating post PDT at Photofrin dose >7 micro g/ml, and at higher dose (28 micro g/ml) plasma membrane disruption and cell swelling were observed immediately after PDT. Dramatic alterations of several important signaling events were detected in A431 cells post Photofrin PDT, including (i) immediate formation of reactive oxygen species (ROS), (ii) rapid activation of c-Jun N-terminal kinase, (iii) delayed activation of caspase 3 and cleavage of polyADP-ribose polymerase and p21-activated kinase 2, and (iv) loss of mitochondrial membrane potential. Intriguingly, the characteristics of typical apoptosis such as phosphatidylserine externalization and DNA fragmentation were not detected in the cell death process caused by this PDT regime. In conclusion, our results show that when plasma membranes are the main targets, Photofrin-PDT can lead to instant ROS formation and subsequent activation of downstream signaling events similar to those elicited by many apoptotic stimuli, but the damage of plasma membranes renders the death phenotype more necrosis like. PMID- 12548557 TI - Cl- channel blockers inhibit transition of quiescent (G0) fibroblasts into the cell cycle. AB - Modulation of ion permeability during the cell cycle is one of the key events in cell cycle progression. We have compared the effects of K+ and Cl- channel blockers on the cell cycle in synchronous and asynchronous NIH3T3 cells. The Cl- channel blocker 5-N-2-(3-phenylpropylamino) benzoic acid (NPPB; 0.2 mM) inhibited entry into S phase in synchronous cells but not in asynchronous cells, while the K+ channel blocker 4-aminopyridine (4-AP) showed similar inhibitory effects in both conditions. In NIH3T3 cells synchronized by serum deprivation/replenishment, G0-to-G1 transition occurred within 8 h after serum addition, and the G1/S checkpoint at 10-14 h. NPPB applied only at 0-8 or 8-14 h after serum addition inhibited entry into S phase. Cl- permeability measured as 125I efflux increased at 4 and 10 h after serum addition. Ki-67-negative cells, which represent quiescent G0 phase cells, progressively decreased in number until 8 h after serum addition. The Cl- channel blockers (NPPB and 4,4'-diisothiocyanatostilbene-2,2' disulfonic acid [DIDS]) but not the K+ channel blocker (4-AP) significantly decreased the rate of reduction in number of Ki-67-negative cells. These data indicate that an increase in Cl- permeability plays an important role in reentry of quiescent cells into the proliferating phase, in addition to the known effects on passage through the G1/S checkpoint. PMID- 12548558 TI - The cytoplasmic tail of invariant chain modulates antigen processing and presentation. AB - The MHC class II-associated invariant chain (Ii) has several important functions in antigen presentation. In this study, we have examined the effect of Iip33 expression on endocytic transport and antigen presentation. We find that degradation of both endocytosed antigen and Ii itself is delayed in cells expressing high levels of Ii, whereas a mutant Ii with an altered charge distribution in the cytoplasmic tail was unable to exert this effect. Furthermore, the Ii mutant did not enhance the presentation of an Ii-dependent MHC class II-restricted epitope to the same extent as the wild type. In a parallel study, we investigated the effect of charge in the cytoplasmic tail of Ii. We find that due to exposed negative charges, it promotes endosome fusion events, and we suggest that this causes endosomal retention (Nordeng et al., Mol. Biol. Cell 2002). Together, the data reveal an additional property of the Iip33 cytoplasmic tail that contributes to the modulation of antigen processing and presentation. PMID- 12548559 TI - T cell recognition and therapeutic effect of a phosphorylated synthetic peptide of the 70K snRNP protein administered in MR/lpr mice. AB - Modifications of self antigens that occur during apoptosis might be involved in the generation of neo-antigens, which can break tolerance and induce autoimmunity. We have previously identified an epitope at residues 131-151 of the U1-70K snRNP protein, recognized by IgG antibodies and CD4+ T cells from at least two strains of lupus mice. With the aim of investigating the possible role of phosphorylation on the antigenicity of peptide 131-151 and to gain a better understanding of how this peptide can drive autoimmune response, we synthesized two peptides phosphorylated on Ser137 and 140, respectively. We show here that peptide P140 phosphorylated on Ser140 is recognized by both CD4+ T cells and antibodies from MRL/lpr mice. Furthermore, intravenous administration to lupus prone MRL/lpr mice of P140 in saline (but not of the non-phosphorylated peptide) decreased proteinuria and anti-DNA antibody production, and significantly prolonged survival of treated mice. We further demonstrated that P140 is recognized by antibodies from lupus patients and binds to various HLA DR molecules, offering new hope for manipulating T cell response in humans. PMID- 12548560 TI - Recirculating and germinal center B cells differentiate into cells responsive to polysaccharide antigens. AB - Antibodies against bacterial capsular polysaccharides play a critical protective role. Responses to these antigens can occur without the help or control of T cells and are associated with marginal zone (MZ) B cells. Capsular antigens are diverse and some cross-react with self-carbohydrate epitopes. This diversity may explain the recruitment of non-autoreactive recirculating B cells and memory B cells to the MZ in addition to other B cells, some of which are weakly autoreactive cells, that are recruited to the MZ without entering the recirculating pool. To test whether memory B cells respond to polysaccharide based antigens, mice with hapten-specific memory B cells were challenged with hapten-polysaccharide. Hapten-specific plasma cells producing high affinity antibody with Ig V-region mutations were induced. To test whether naive recirculating B cells can form MZ cells that respond to polysaccharide, recirculating B cells from lymph nodes were transferred into Rag-1-deficient mice. MZ cells differentiated from the donor cells without proliferation or T cell help and responded to polysaccharide-based antigen. The differentiation of B cells both from germinal centers and the recirculating pool to the MZ phenotype is likely to make an important contribution to the repertoire of B cells that respond to polysaccharide antigens. PMID- 12548561 TI - The interaction between NK cells and dendritic cells in bacterial infections results in rapid induction of NK cell activation and in the lysis of uninfected dendritic cells. AB - NK and DC reciprocal interactions have only recently been investigated. In this study, we focused on the interplay between NK cells and DC in two models of bacterial infection. Immature monocyte-derived DC were cultured in the presence of live Escherichia coli or bacillus Calmette-Guerin. Upon exposure to either extracellular or intracellular bacteria, DC underwent maturation as assessed by the increased levels of expression of CD80,CD86, and HLA molecules and the de novo expression of CD83 and CCR7. Significant amounts of TNF-alpha and IL-12 were released by DC upon infection, whereas IL-2 and IL-15 were barely detectable in culture supernatants. Both infected and uninfected DC were capable of inducing in fresh autologous NK cells the expression of CD69 and HLA-DR and of inducing cell proliferation. Remarkably, however, infected DC were much stronger inducers of NK cell activation and proliferation than uninfected DC. Thus, after just 24 h of NK/DC coculture, only those NK cells that had been exposed to bacteria-infected DC had acquired the ability to lyse autologous immature DC. In addition, infected DC were more resistant to NK-mediated lysis as a consequence of the up-regulation of HLA class I molecule expression on their surface. This study suggests a regulatory circuit involving NK cells and DC in which DC-induced NK cell activation is effectively enhanced by the presence of pathogens. Activated NK cells, by limiting the supply of immature DC, may then exert a control on subsequent innate and adaptive immune responses. PMID- 12548562 TI - Transgenic mice with hematopoietic and lymphoid specific expression of Cre. AB - Bacteriophage P1 Cre/loxP based systems can be used to manipulate the genomes ofmice in vivo and in vitro, allowing the generation of tissue-specific conditional mutants. We have generated mouse lines expressing Cre recombinase in hematopoietic tissues using the vav regulatory elements, or in lymphoid cells using the hCD2 promoter and locus control region (LCR). The R26R-EYFP Cre reporter mouse line was used to determine the pattern of Cre expression in each line and enabled the assessment of Cre activity at a single-cell level. Analysis showed that the vav promoter elements were able to direct Cre-mediated recombination in all cells of the hematopoietic system. The hCD2 promoter and LCR on the other hand were able to drive Cre-mediated recombination only in T cells and B cells, but not in other hematopoietic cell types. Furthermore, in the appropriate tissues, deletion of the floxed target was complete in all cells, thereby excluding the possibility of variegated expression of the Cre transgene. Both of these Cre-transgenic lines will be useful in generating tissue-specific gene deletions within all the cells of hematopoietic or lymphoid tissues. PMID- 12548565 TI - Molecular characterization of the novel rat NK receptor 1C7. AB - A novel receptor, named 1C7 or NKp30 and involved in natural cytotoxicity, was recently identified. This receptor is encoded by the 1C7 gene, which is located within the class III region of the human MHC, HLA. It is a member of the immunoglobulin gene superfamily (IgSF) and, remarkably, is expressed at the mRNA level as six different splice variants in human. Recent investigations have indicated that the 1c7 gene of the mouse is silenced by in-frame stop codons. In this study, the molecular characterization of the rat 1c7 gene is described. cDNA derived from this gene encode a protein of 192 amino acid residues predicted to contain a single IgV-set domain in the extracellular region and a positively charged residue in the transmembrane region. Expression of the gene was detected in freshly isolated rat Natural Killer (NK) and T splenocytes. Transfection of rat 1C7 into the NK cell line RNK-16 induced cytolytic activity against glioma as well as lymphoma tumor cells. In addition, binding of a r1C7-Fc fusion protein by a panel of target cells correlated with susceptibility to killing by RNK-16-1C7 effector cells. These results indicate that the r1C7 molecule could function as an NK activating receptor as previously reported for the human NKp30 receptor molecule. PMID- 12548563 TI - Outer membrane protein A renders dendritic cells and macrophages responsive to CCL21 and triggers dendritic cell migration to secondary lymphoid organs. AB - Outer membrane protein A (OmpA) is a class of bacterial cell wall protein that is immunogenic without adjuvant. As specific immune responses are initiated in the lymph nodes (LN, we analyzed the effect of the OmpA from Klebsiella pneumoniae (KpOmpA) onchemokine/ chemokine receptor expression by APC and on cell migration to the LN. Upon contact with KpOmpA, human immature DC and macrophages acquire CCR7 expression and responsiveness to CCL21. In parallel, CCR1 and CCR5 expression is down-regulated and CXCL8, CCL2, CCL3 and CCL5 production is up regulated. Mice injected subcutaneously with KpOmpA present a transient inflammatory reaction at the site of injection accompanied by an enlargement of the draining LN with a higher proportion of DC and macrophages. Lastly, when exposed to KpOmpA prior injection, DC but not macrophages migrate to the draining LN. In conclusion, KpOmpA confers a migratory phenotype to DC and triggers their migration to the regional LN. This property contributes to explain how innate cells initiate adaptive immune response upon recognition of conserved bacterial components and also why OmpA is immunogenic in the absence of adjuvant. PMID- 12548564 TI - TRANCE together with IL-7 induces pre-B cells to proliferate. AB - TRANCE (TNF-related activation-induced cytokine)-deficient mice completely lack osteoclasts, and develop severe osteopetrosis. These mice also show a defect in their pre-B cell differentiation. In the present study, the expression of TRANCE was examined in pre-B cell lines using flow cytometry and reverse transcription PCR. Three pre-B cell lines, 18-81, B3P816-1, and 38B9, expressed TRANCE on their surface, and two pre-B cell lines, 7OZ/3 and NFS5, at the late pre-B cell stage, expressed it at low levels, although their mRNA expression was normal. Another pre-B cell line, 38-C-13, at the intermediate stage between pre-B and immature B cells, did not express TRANCE. The IL-7-dependent pre-B cell line PreBR, which expresses the pre-B cell receptor on the cell surface, also expressed TRANCE. When differentiation of PreBR cells was induced in vitro by removing IL-7 from cultures, TRANCE expression dropped; it was restored by the addition of IL-7, suggesting that TRANCE functions in cooperation with IL-7. To examine the function of TRANCE, we introduced the TRANCE gene into PreBR cells and established two transfectants that constitutively expressed TRANCE, even in the absence of IL-7. In these transfectants, after removal of IL-7, the number of cells that succeeded in kappa chain rearrangement was decreased to one third; and CD40 expression decreased to less than one tenth. Moreover, the percentage of cells in the S/G2/M phase was increased by 50% over the mock transfectant. These findings indicate that, before kappa chain rearrangement occurs, TRANCE together with IL-7 induces pre-B cells to proliferate and makes this rearrangement more efficient. PMID- 12548566 TI - Somatic hypermutation does not require Rad54 and Rad54B-mediated homologous recombination. AB - Secondary diversification of immunoglobulin (Ig) genes occurs through somatic hypermutation (SHM) in B cells of the germinal center (GC). The GC reaction is associated with a high frequency of DNA double-strand breaks (DSB) in the hypermutation domain of Ig genes. Homologous recombination (HR) is a prominent DSB repair pathway. Among the proteins involved in HR are the Rad-54 paralogues, Rad54 and Rad54B. To investigate whether Rad54/Rad54B-mediated HR is involved in SHM, we determined the ratio of mutated versus non-mutated Vlambda PCR products from memory (IgM-, IgD-, Vlambda1+) and GC (PNA(high), Vlambda1+) B cells, the mutation load, the mutation frequency, the base exchange pattern and the distribution of somatic mutations along the rearranged Vlambda light chain (VlambdaLC) genes. All these parameters of SHM were unaltered in memory and GC B cells lacking one or both Rad54 paralogues. Thus, our data indicate that Rad54 and Rad54B-mediated HR is not essential for SHM. In addition, the finding that the ablation of RAD51 paralogues causes an increase in SHM argues against a direct involvement of HR in promoting SHM. PMID- 12548567 TI - CD2+/CD14+ monocytes rapidly differentiate into CD83+ dendritic cells. AB - Since denditric cells (DC) represent the main players linking innate and adaptive immunity, their prompt generation from blood cells would be instrumental for an efficient immune response to infections. Consistent with this, CD2+ monocytes were found to express the DC maturation marker CD83, along with acquisition of high antigen-presenting activity, after a surprisingly short time in culture. This rapid process is associated with expression of IFN-alpha/beta genes and secretion of low levels of pro-inflammatory cytokines. Exposure of monocytes to IFN-alpha, but not to IL-4, induced persistence of CD2+/CD83+ cells, which were fully competent in stimulating primary responses by naive T cells. These results unravel the natural pathway by which infection-induced signals rapidly transform pre-armed monocytes into active DC. PMID- 12548568 TI - Third North American Symposium on Skeletal Complications of Malignancy: summary of the scientific sessions. AB - The current introduction summarizes the five scientific sessions (bone marrow microenvironment and animal models of bone metastasis; prostate carcinoma; multiple myeloma; breast carcinoma; and preclinical and clinical studies on metastasis: future directions) and provides an overview of the proceedings of the Third North American Symposium on Skeletal Complications of Malignancy, Bethesda, Maryland, April 25-27, 2002. PMID- 12548569 TI - The development and function of the skeleton and bone metastases. AB - Bone is a frequent site of metastases of the most common tumors, e.g., breast carcinoma and prostate carcinoma. The functions of the skeleton, calcium homeostasis and mechanical support, are carried out by the continuous destruction and rebuilding of small packets of this tissue called bone remodeling. Multinucleated, hemopoietically derived osteoclasts, which are related to macrophages, digest the bone, and mesenchymal-derived osteoblasts rebuild it. This process is kept in balance by finely regulated processes whereby osteoblast lineage cells respond to homeostatic signals and release factors that regulate osteoclast generation and activity. Cells that participate in inflammation and immunity also can stimulate osteoclast formation and lead to bone destruction. Tumor cells most likely subvert these physiologic processes to lodge in bone and cause metastases. PMID- 12548570 TI - Role of stromal-derived cytokines and growth factors in bone metastasis. AB - BACKGROUND: Interactions between bone marrow-derived cytokines, growth factors, and tumors play a critical role in both the homing of tumors to the bone and the development of bone metastasis. Bone is a storehouse of latent growth factors produced by stromal cells and osteoblasts that, when activated during osteoclastic bone resorption, can enhance the growth of tumor cells. METHODS: This article reviews the role these factors may play in bone metastasis. RESULTS: Several studies have shown that breast carcinoma cells, which induce osteoclastic bone resorption, release growth factors that enhance tumor growth. In addition, bone-derived growth factors and chemokines, such as stromal cell-derived factor 1 and monocyte chemoattractant protein 1, can act as chemoattractants to attract tumor cells to bone. Finally, the interaction between tumor cells and bone marrow stromal cells can result in increased production of cytokines and growth factors, such as interleukin 6 or the ligand for the receptor activator of nuclear factor kappaB, that can enhance bone destruction, tumor growth, and angiogenesis. CONCLUSIONS: Stromal cell-derived cytokines and growth factors as well as growth factors that are released during the bone resorption process play a critical role in the development of bone metastasis. Interruption of this symbiotic relation between tumors that induce bone destruction and release of bone-derived growth factors can have beneficial effects on blocking both bone destruction and decreasing tumor burden within bone. PMID- 12548571 TI - Stromal factors involved in prostate carcinoma metastasis to bone. AB - BACKGROUND: Prostate carcinoma (PC) frequently metastasizes to bone, where it causes significant morbidity and mortality. Stromal elements in the primary and metastatic target organs are important mediators of tumor cell intravasation, chemoattraction, adhesion to target organ microvascular endothelium, extravasation, and growth at the metastatic site. METHODS: The role of stromal factors in bone metastasis was determined with a cyclic DNA microarray comparison of a bone-derived cell PC cell line with a soft tissue-derived cell PC cell line and by evaluating the effects of selected stromal components on PC cell chemotaxis, cell adhesion to human bone marrow endothelium (HBME), and PC cell growth. RESULTS: The authors demonstrate that PC cells express protease-activated receptor 1 (PAR1; thrombin receptor), and its expression is up-regulated in PC compared with normal prostate tissue. In addition, this overexpression was very pronounced in bone-derived PC cell lines (VCaP and PC-3) compared with soft tissue PC cell lines (DUCaP, DU145, and LNCaP). The authors report that bone stromal factors, including stromal cell-derived factor 1 (SDF-1) and collagen Type I peptides, are chemoattractants for PC cells, and they demonstrate that some of these factors (e.g., extracellular matrix components, transforming growth factor beta, bone morphogenic proteins [BMPs], and SDF-1) significantly alter PC HBME interaction in vitro. Finally, stromal factors, such as BMPs, can regulate the proliferation of PC cells in vitro. CONCLUSIONS: Soluble and insoluble elements of the stroma are involved in multiple steps of PC metastasis to bone. The authors hypothesize that PAR1 may play a central role in prostate tumorigenesis. PMID- 12548572 TI - Animal models of bone metastasis. AB - BACKGROUND: Animal models are important tools to investigate the pathogenesis and develop treatment strategies for bone metastases in humans. However, there are few spontaneous models of bone metastasis despite the fact that rodents (rats and mice) and other animals (dogs and cats) often spontaneously develop cancer. Therefore, most experimental models of bone metastasis in rodents require injection or implantation of neoplastic cells into orthotopic locations, bones, or the left ventricle of the heart. METHODS: The current study reviews the natural incidence and clinical manifestation of bone metastases of mammary and prostate carcinoma in animals, as well as the experimental models developed in mice using animal and human-derived neoplasms. RESULTS: Rats, mice, dogs, and cats often develop spontaneous mammary carcinoma, but bone metastases are rare. Intact and neutered dogs develop prostate carcinoma that is usually androgen independent and may be associated with regional bone invasion or distant bone metastasis. Normal dog prostate tissue induces new bone formation in vivo and can serve as a model of osteoblastic metastasis without concurrent bone destruction. Experimental models of osteolytic, osteoblastic, and mixed osteolytic/osteoblastic bone metastases include syngeneic rodent neoplasms or human xenografts implanted at orthotopic sites (e.g., breast or prostate glands) in immunodeficient mice, injection of cancer cells into the left ventricle of the heart, or direct injection into bones. New transgenic mouse models of cancer have a low incidence of spontaneous bone metastasis, but cell lines derived from these tumors can be selected in vivo for increased incidence of bone metastasis. It is essential to validate and correctly interpret the lesions in models of bone metastasis to accurately correlate the data from animal models to human disease. Animal models have provided support for the "seed and soil" hypothesis of bone metastasis. However, the roles of vascular patterns in the metaphyses of long bones and rapid bone turnover in young animals in the pathogenesis of metastasis in experimental models are uncertain. Improvements in the imaging of experimental animals in vivo using fluorescent markers or light emitted from luciferase have led to increased sensitivity of detection and more accurate quantification of bone metastases. For example, imaging of human prostate carcinoma PC-3M cells transfected with luciferase, following injection into the left ventricle, has demonstrated that there is rapid localization of tumor cells to bones and other organs, such as the kidneys and lungs. CONCLUSIONS: Animal models of metastasis have supported drug development and have been useful for identification of metastasis suppressor and promoter genes as novel targets for the development of novel therapies. Further refinement of these models will involve spatiotemporal analysis of the metastatic process by imaging and use of image data to stage disease and guide tissue sampling for gene expression profiling via gene array technology. In the future, integrated analyses of these models will be needed to understand the complexities of this important disease process. PMID- 12548573 TI - Prostate carcinoma: defining therapeutic objectives and improving overall outcomes. AB - The development and progression of a prostate carcinoma from prediagnosis to death can be characterized as a series of clinical states. The states are milestones that can be used to assess prognosis, define therapeutic objectives, and assess outcomes. The antitumor effects of hormone therapies and cytotoxic agents in patients with prostate carcinoma are placed in context along with the bidrectional tumor-host interactions that contribute to the growth and resistance of osseous lesions. Identifying the factors that contribute to the growth of the disease at different points in the illness has lead to novel, systemic approaches. Proving the benefit of these approaches requires a series of unique trials with unique endpoints relevant to the clinical state of the patients and the specific therapy under evaluation. PMID- 12548574 TI - Prostate carcinoma bone-stroma interaction and its biologic and therapeutic implications. AB - BACKGROUND: The homing of prostate carcinoma to bone is a nonrandom, multistep process. Previous studies have revealed significant insights into how tumor cells can interact with the host microenvironment. In this communication, the author summarizes recent studies from his institution and draws conclusions from data published by others pertaining to the biologic and therapeutic implications of bone metastasis from prostate carcinoma. METHODS: Tumor models have been established to study cellular interaction between human prostate carcinoma cells and bone stroma under two-dimensional and three-dimensional conditions. At the author's institution, experiments were conducted to show that prostate carcinoma cell growth and survival are enhanced in coculture with pleuripotent bone stromal cells. A cotargeting concept for the treatment of patients with prostate carcinoma bone metastasis is introduced. RESULTS: Both genotypical and phenotypical responses were observed to tumor epithelium when it was cocultured under three-dimensional conditions. A "vicious cycle" that was mediated by soluble and insoluble molecules secreted by tumor and bone may be the key to supporting and sustaining tumor colonization in bone. Cotargeting tumor and stroma has yielded promising results, both in preclinical models of prostate carcinoma bone metastasis and in the clinic with patients who were treated with a dual tumor-targeting and bone-targeting strategy. CONCLUSIONS: Understanding and targeting the interaction of tumor cells and bone stroma may improve the prognosis, reduce the suffering, and increase the survival of patients with advanced bone metastasis as a consequence of prostate carcinoma. PMID- 12548575 TI - Role of endothelin-1 in osteoblastic bone metastases. AB - BACKGROUND: Certain solid tumors metastasize to bone and cause an osteoblastic response. The mechanisms by which tumor cells stimulate this new bone formation are not completely understood. METHODS: The authors identified three breast cancer lines that cause osteoblastic metastases in female nude mice and provided evidence that tumor-produced endothelin-1 (ET-1) mediates the osteoblastic response. RESULTS: Tumor conditioned media, as well as exogenous ET-1, stimulated osteoblast proliferation and new bone formation in cultures of mouse calvariae. These effects were blocked by antagonists of the endothelin A (ET(A)), but not ET(B), receptors. Mice inoculated with the ZR-75-1 breast cancer line and treated with a selective ET(A) receptor antagonist (ABT-627) had significantly fewer osteoblastic bone metastases and less tumor burden compared with untreated mice. In contrast, there was no effect of ABT-627 on osteolytic bone metastases caused by ET-1-negative breast cancer, MDA-MB-231. ABT-627 had no effect on growth in vitro or at the orthotopic site of ZR-75-1 or MDA-MB-231 cells. CONCLUSIONS: Collectively, the data suggested that tumor-produced ET-1 mediates osteoblastic bone metastases by stimulating osteoblast proliferation and new bone formation. ET(A) receptor blockade may be useful for prevention and the treatment of osteoblastic bone metastases due to breast or prostate cancer. PMID- 12548576 TI - Targeting prostate cancer bone metastases. PMID- 12548577 TI - Diagnosis and management of treatment-related osteoporosis in men with prostate carcinoma. AB - Osteoporosis is a complication of androgen deprivation therapy in men with prostate carcinoma. Androgen deprivation therapy, caused by either bilateral orchiectomy or treatment with a gonadotropin-releasing hormone agonist, decreases bone mineral density and increases fracture risk. Other factors including diet and lifestyle may contribute to bone loss. There is limited information regarding the best strategy to prevent osteoporosis in men with prostate carcinoma. Lifestyle modification including smoking cessation, moderation of alcohol consumption, and regular weight-bearing exercise should be encouraged. Supplemental calcium and vitamin D are also recommended. Additional treatment may be warranted for men with osteoporosis, fractures, or high rates of bone loss during androgen deprivation therapy. Intravenous pamidronate, a second-generation bisphosphonate, prevents bone loss during androgen deprivation therapy. Zoledronic acid, a more potent third-generation bisphosphonate, not only prevents bone loss but also increases bone mineral density during androgen deprivation therapy. Other bisphosphonates may be effective although they have not been evaluated in this clinical setting. Treatment with estrogens or selective estrogen receptor modulators may also be effective. Monotherapy with bicalutamide or other antiandrogens may cause less bone loss than androgen deprivation therapy. PMID- 12548578 TI - Moving disease biology from the lab to the clinic. AB - Multiple myeloma (MM) cells home to and adhere to extracellular matrix proteins and to bone marrow stromal cells (BMSCs); and in the BM microenvironment, grow, survive, resist drugs, and migrate under the influence of cytokines including interleukin-6, vascular endothelial growth factor, tumor necrosis factor alpha, and insulin-like growth factor (IGF)-1. Proliferation is via the Ras/Raf MAPK cascade, drug resistance via PI3-K/Akt signaling, and migration via PKC dependent pathways. Novel therapies that target not only the MM cell, but also the BM microenvironment, can overcome drug resistance in vitro and in vivo in murine human MM models. For example, immunomodulatory derivatives of thalidomide (IMiDs) and the proteasome inhibitor PS-341 both induce apoptosis of MM cell lines and patient cells refractory to melphalan, doxorubicin, and dexamethasone; abrogate MM cell binding to fibronectin and BMSCs and related protection against immune- and drug-induced apoptosis; block production of cytokines which promote MM cell growth, survival, drug resistance, and migration; inhibit angiogenesis; and stimulate host anti-tumor immunity. In the setting of relapsed refractory MM, a Phase I trial of the IMiD CC5013 shows stable paraprotein or better in 20 of 24 (79%) patients, with a favorable toxicity profile. In this same patient population 85% of 54 patients treated in a Phase II trial of PS-341 achieved either paraprotein response (50%) or stable disease (35%). Cellular and gene microarray studies comparing PS-341 and an IkappaB kinase inhibitor, PS-1145, suggest that selective NF-kappaB blockade cannot account for all the anti-MM activity of PS-341. Finally, cellular and signaling studies provide the preclinical rationale for combining these novel agents with conventional therapies, or with each other, to enhance efficacy. These novel therapeutics therefore represent a new treatment paradigm in MM targeting the tumor cell in its microenvironment to overcome classical drug resistance and improve patient outcome. Future studies should define the utility of these agents as primary therapy, treatment for first relapse, and maintenance therapy. PMID- 12548579 TI - RANK-Fc: a therapeutic antagonist for RANK-L in myeloma. AB - BACKGROUND: Severe bone destruction due to inappropriate osteoclastogenesis is a prominent feature of multiple myeloma (MM). MM increases bone loss by disrupting the checks that normally control signaling by receptor activator of nuclear factor kappaB ligand (RANK-L, also called TRANCE [tumor necrosis factor-related, activation-induced cytokine], osteoprotegerin ligand [OPG-L], osteoclast differentiation factor [ODF], and tumor necrosis factor superfamily member 11 [TNFSF11]), a TNF-family cytokine required for osteoclast differentiation and activation. RANK-L binds to its functional receptor RANK (TNF receptor superfamily member 11a [TNF RSF11a]) to stimulate osteoclastogenesis. Osteotropic cytokines regulate this process by controlling bone marrow stromal expression of RANK-L. Further control over osteoclastogenesis is maintained by regulated expression of osteoprotegerin (OPG, also called osteoclastogenesis inhibitory factor and TNFRSF11b), a soluble decoy receptor for RANK-L. In normal bone marrow, abundant stores of OPG in stroma, megakaryocytes, and myeloid cells provide a natural buffer against increased RANK-L. MM disrupts these controls by increasing expression of RANK-L and decreasing expression of OPG. Concurrent deregulation of RANK-L and OPG expression is found in bone marrow biopsies from patients with MM but not in specimens from patients with non-MM hematologic malignancies. METHODS: RANK-Fc is a recombinant RANK-L antagonist that is formed by fusing the extracellular domain of RANK to the Fc portion of human immunoglobulin G(1) (hIgG(1)). In vitro, addition of RANK-Fc virtually eliminates the formation of osteoclasts in cocultures of MM with bone marrow and osteoblast/stromal cells. The severe combined immunodeficiency (SCID)/ARH77 mouse model and the SCID-hu-MM mouse model of human MM were used to assess the ability of RANK-Fc to block the development of MM-induced bone disease in vivo. Mice received either RANK-Fc or hIgG(1) 200 microg intravenously three times per week. RESULTS: RANK-Fc limited bone destruction in both the SCID/ARH-77 model and the SCID-hu-MM model. Administration of RANK-Fc also caused a marked reduction in tumor burden and serum paraprotein in SCID-hu-MM mice that was associated with the restoration of OPG and a reduction in RANK-L expression in the xenograft. CONCLUSIONS: MM-induced bone destruction requires increased RANK-L expression and is facilitated by a concurrent reduction in OPG, a natural decoy receptor for RANK-L. Administration of the RANK-L antagonist RANK-Fc limits MM-induced osteoclastogenesis, development of bone disease, and MM tumor progression. PMID- 12548580 TI - Receptor activator of NF-kappaB ligand, macrophage inflammatory protein-1alpha, and the proteasome: novel therapeutic targets in myeloma. AB - BACKGROUND: The bone destruction in myeloma patients is largely responsible for the clinical features of the disease. However, only recently has attention focused on identifying and developing drugs targeted specifically at the osteolysis. Receptor activator of NF-kappaB ligand (RANKL), macrophage inflammatory protein (MIP)-1alpha, and proteasomal function have been implicated in the pathogenesis of myeloma and associated bone disease. We provide "proof of principle" in preclinical myeloma models that these are indeed valid molecular targets in development of novel therapeutics. METHODS: The efficacy of antagonists of RANKL and MIP-1alpha bioactivities (RANK.Fc and neutralizing monoclonal anti-MIP-1alpha antibody) in ameliorating osteolysis and reducing tumor burden was evaluated in a mouse model in which murine myeloma 5TGM1 cells are injected intravenously into syngeneic mice. In addition, the activity of a petidyl aldehyde proteasome inhibitor (proteasome inhibitor-1 [PSI]) on tumor growth was tested in a murine 5TGM1 plasmacytoma model and in mice intravenously inoculated with 5TGM1 cells. RESULTS: RANK.Fc and anti-MIP-1alpha antibody inhibited the development and progression of osteolytic lesions and significantly reduced tumor load assessed by serum monoclonal paraprotein titers. Intratumoral injections of PSI inhibited growth of 5TGM1 plasmacytomas and induced tumor regression in some cases. In addition, systemic administration of PSI significantly prolonged time to onset of paraplegia in tumor-bearing mice. CONCLUSIONS: The results highlight the critical roles of RANKL and MIP-1alpha in the development and progression of myeloma and provide a basis for future evaluation in myeloma patients of novel therapeutics that disrupt interactions of RANKL and MIP-1alpha with their cognate receptors. The data also suggest that further studies in preclincal myeloma models aimed at identifying other proteasome inhibitors with antitumor efficacy would be worthwhile. PMID- 12548581 TI - Bisphosphonates and osteoprotegerin as inhibitors of myeloma bone disease. AB - BACKGROUND: A major clinical feature in multiple myeloma is the development of osteolytic bone disease. The increase in bone destruction is due to uncontrolled osteoclastic bone resorption. Until recently the factors responsible for mediating the increase in osteoclast formation in myeloma have been unclear. However, recent studies have implicated a number of factors, including the ligand for receptor activator of NFkappaB (RANKL) and macrophage inflammatory protein 1alpha. The demonstration that increased osteoclastic activity plays a central role in this process and the identification of molecules that may play a critical role in the development of myeloma bone disease have resulted in studies aimed at identifying new approaches to treating this aspect of myeloma. METHODS: Studies have been performed to determine the ability of recombinant osteoprotegerin (Fc.OPG), a soluble decoy receptor for RANKL, and potent new bisphosphonates to inhibit the development of myeloma bone disease in the 5T2MM murine model of multiple myeloma. RESULTS: Fc.OPG was shown to prevent the development of osteolytic bone lesions in 5T2MM bearing animals. These changes were associated with a preservation of the cancellous bone loss induced by myeloma cells and an inhibition of osteoclast formation. Bisphosphonates, including ibandronate and zoledronic acid, were also shown to inhibit the development of osteolytic bone lesions in the 5T2MM model and alternative models of myeloma bone disease. CONCLUSIONS: Bisphosphonates and Fc.OPG are effective inhibitors of the development of osteolytic bone lesions in pre-clinical murine models of myeloma bone disease. PMID- 12548582 TI - The biology of breast carcinoma. AB - The biology of breast carcinoma is complex, with multiple factors contributing to its development and progression. The current review focuses on the role of several critical genes including estrogen receptor, progesterone receptor, retinoic acid receptor-beta, epidermal growth factor receptor family members, p53, BRCA1, and BRCA2 as risk factors for the development of disease, predictors of prognosis and response to therapy, and as therapeutic targets. Studies of the biology of these and other genes that contribute to the development and progression of breast carcinoma have had and will continue to have great impact on all aspects of disease management. PMID- 12548583 TI - Mechanisms of osteolytic bone metastases in breast carcinoma. AB - Osteolytic and osteoblastic metastases are often the cause of considerable morbidity in patients with advanced prostate and breast carcinoma. Breast carcinoma metastasis to bone occurs because bone provides a favorable site for aggressive behavior of metastatic cancer cells. A vicious cycle arises between cancer cells and the bone microenvironment, which is mediated by the production of growth factors such as transforming growth factor beta and insulin growth factor from bone and parathyroid hormone-related protein (PTHrP) produced by tumor cells. Osteolysis and tumor cell accumulation can be interrupted by inhibiting any of these limbs of the vicious cycle. For example, bisphosphonates (e.g., pamidronate, ibandronate, risedronate, clodronate, and zoledronate) inhibit both bone lesions and tumor cell burden in bone in experimental models of breast carcinomametastasis. Neutralizing antibodies to PTHrP, which inhibit PTHrP effects on osteoclastic bone resorption, also reduce osteolytic bone lesions and tumor burden in bone. Other pharmacologic approaches to inhibit PTHrP produced by breast carcinoma cells in the bone microenvironment also produce similar beneficial effects. Identification of the molecular mechanisms responsible for osteolytic metastases is crucial in designing effective therapy for this devastating complication. PMID- 12548584 TI - Antitumor effects of bisphosphonates. AB - BACKGROUND: Bisphosphonates are widely used to treat skeletal complications of malignancy. These drugs accumulate in bone where they inhibit osteoclastic bone resorption and reduce the local release of factors that stimulate tumor growth. The mechanism of action of bisphosphonates is dependent on chemical structure: Nonnitrogen-containing compounds (e.g., etidronate, clodronate) are metabolized into cytotoxic analogues of ATP, whereas the more potent nitrogen-containing compounds (N-BPs; e.g., pamidronate, ibandronate, zoledronic acid) inhibit protein prenylation, thus affecting cell function and survival. Because protein prenylation is required by all cells, not just osteoclasts, the possibility arises that N-BPs could also affect the viability of tumor cells. METHODS: Several groups have investigated the in vitro effects of bisphosphonates, either alone or in combination with other antineoplastic agents, on the viability and metastatic properties of many tumor cell types. Similarly, the effect of bisphosphonate treatment on osteolysis and tumor burden has been studied in a variety of animal tumor models. RESULTS: In vitro, submicromolar concentrations of N-BPs inhibited tumor cell adhesion and reduced invasion through extracellular matrix. At higher concentrations, antiproliferative and proapoptotic effects have been reported. In animal models of bone metastases, bisphosphonate treatment markedly reduced osteolytic lesions. There is also evidence of a reduction in tumor burden in bone and occasionally in other organs. Survival may be prolonged, but bisphosphonates do not appear to inhibit the growth of primary soft tissue tumors or orthotopic xenografts. CONCLUSIONS: The cell culture data clearly demonstrated that N-BPs exert antitumor properties and interact synergistically with other antineoplastic agents. As bisphosphonates accumulate in bone, they can also exert cytostatic effects on tumor cells in bone metastases, either directly or indirectly via osteoclast inhibition and alterations in the bone microenvironment. Further in vivo research is now required to optimize the dosing regimen of N-BPs to exploit fully their antitumor potential. PMID- 12548585 TI - Bisphosphonates and metastatic breast carcinoma. AB - BACKGROUND: Metastasis to the skeleton occurs in approximately 75% of patients with advanced breast cancer. It is the cause of much morbidity and decreased quality of life in these patients. METHODS: Zoledronic acid (Zoledronate) is a new, potent third-generation bisphosphonate that has recently been approved by the U.S. Food and Drug Administration for the treatment of both osteolytic and osteoblastic bone metastases. A Phase 3 clinical trial comparing the efficacy of Zoledronic acid 4 and 8 mg compared with Pamidronate 90 mg every 3-4 weeks has been completed. RESULTS: A 15 minute infusion of 4 mg of zoledronic acid is at least as effective as 90 mg of pamidronate in preventing skeletal complications in patients with metastatic breast cancer. CONCLUSIONS: Bisphosphonate treatment can delay skeletal events in patients with breast carcinoma that is metastatic to bone. Future trials will build on the results achieved with bisphosphonate treatment. PMID- 12548586 TI - Bisphosphonate therapy for patients with breast carcinoma. AB - Bisphosphonates have become well established in the treatment of patients with metastatic bone disease, although the optimal use of these agents has not been defined clearly. Randomized, controlled trials have demonstrated that treatment with intravenous pamidronate can significantly reduce the rate of skeletal related events in patients with bone metastases from myeloma or advanced breast carcinoma. To date, there are few data from controlled, randomized studies to support the use of bisphosphonates in patients with bone metastases from malignancies other than breast carcinoma and myeloma. The optimal duration of treatment is unknown. Recent data have demonstrated that prolonged treatment is tolerated well, with no obvious toxicity. Generally, treatment is continued irrespective of the development of skeletal-related events and until there is a substantial decline in performance status. The widespread use of bisphosphonates will have major financial implications. Retrospective studies have suggested that the cost-effectiveness ratio is high for patients with advanced breast carcinoma. These ratios may be improved by targeting therapy to patients at high risk of developing complications from skeletal metastatic disease. Among patients with skeletal metastases from breast carcinoma, a recent retrospective analysis demonstrated that patients with disease confined to the skeleton were at greater risk of pathologic fractures compared with patients who had additional extraosseous disease. It is interesting to note that approximately two-thirds of patients with advanced breast carcinoma in the randomized trials of intravenous pamidronate had disease confined to the skeleton. The use of markers of bone turnover to identify patients who are most likely to benefit from bisphosphonate therapy or to identify patients who will respond to such therapy is the subject of further investigation. There are conflicting data on the use of bisphosphonates as an adjuvant therapy. Currently, such treatment should occur only as part of a clinical trial. Bisphosphonates can be used to prevent bone loss as a result of therapy for malignant disease, e.g., premature menopause in patients with early breast carcinoma. PMID- 12548587 TI - Effectiveness and cost of bisphosphonate therapy in tumor bone disease. AB - BACKGROUND: Tumor-induced osteolysis due to breast carcinoma and myeloma is responsible for a considerable morbidity that severely impairs patients'quality of life. Osteoclast-mediated bone resorption is reported to be increased markedly in patients with tumor bone disease and can be inhibited by bisphosphonate therapy. METHODS: The incidence of skeletal complications and the effectiveness of bisphosphonate therapy in patients with breast carcinoma metastatic to bone or in those with myeloma were derived from large-scale, long-term, placebo controlled trials with clodronate or pamidronate. To the authors' knowledge, there are few studies published to date evaluating the cost-effectiveness of bisphosphonate therapy, and the majority that do exist often are based on models and are applicable only to a particular health care system. RESULTS: From the placebo groups of the above-mentioned trials, one can estimate that approximately 25-40% of the patients with breast carcinoma metastatic to bone will require radiotherapy for bone pain and approximately 17-50% will sustain incident vertebral fractures yearly. The incidence of complications is reported to be lower in myeloma patients. The prolonged administration of bisphosphonates reportedly can reduce the frequency of skeletal-related events by approximately 25-50%. Maximal efficacy appears to have been achieved with the current therapeutic schemes based on monthly intravenous infusions. Beneficial effects appear to be obtained more readily using the intravenous route rather than the oral route. The costs of bisphosphonate therapy appear to be higher than the cost savings from the prevention of skeletal-related events. The costs per quality of life-adjusted year have been estimated to be > $100,000, but more research is needed. Limited data suggest that zoledronic acid will not reduce treatment costs but the short infusion time will lead to substantial time savings for patients and for outpatient oncology facilities. CONCLUSIONS: As is the case for many agents used in oncology, bisphosphonates remain a relatively expensive therapy. More studies are needed to evaluate their cost-effectiveness ratio correctly. A ceiling effect has been reached with current therapeutic schemes and tailoring therapy to the individual patient needs to be evaluated correctly to increase therapeutic effectiveness and improve quality of life further without increasing treatment costs. PMID- 12548588 TI - Bone cancer pain. AB - BACKGROUND: Bone cancer pain is very common, and patients with this type of pain may be difficult to treat. Development of an experimental model for studying this condition is critical to advancing an understanding of the mechanisms that cause pain in patients with malignant disease. METHODS: A murine model of bone cancer was studied. Combined analysis of the extent of tumor-induced bone destruction, pain, and neurochemical characterization of the peripheral and central nervous systems was performed to investigate bone cancer pain. Disease-induced bone destruction was assessed by radiographs and histomorphometry. Pain was assessed by spontaneous and elicited behaviors, and neurochemical analysis involved immunohistochemical detection of hyperalgesic peptides and neurochemical markers. RESULTS: Mice with distal femoral sarcomas exhibited behavioral and neurochemical measures of pain. The pain condition created by malignant bone disease was distinct neurochemically from inflammatory and neuropathic pain states. Experimental evidence indicated that both disease-induced osteolysis and tumors themselves contributed to the generation of pain and that peripheral and central sensitization of the nervous system was present. CONCLUSIONS: Malignant bone disease creates a unique pain state that involves sensitization of the nervous system. Major contributors to the pain state within the bone tissue are osteoclastic bone resorption and the malignant disease itself. PMID- 12548589 TI - Bone markers in the management of patients with skeletal metastases. AB - BACKGROUND: Biochemical markers of bone remodeling have been used as surrogate markers to manage patients with metastatic bone disease. Markers of the bone resorptive process, such as the bone collagen breakdown products N-telopeptide and C-telopeptide, are useful markers for monitoring the response and efficacy of antiresorptive therapy and to assess disease progression in patients with osteolytic bone disease. Recently discovered markers of osteoclastogenesis, osteoprotegerin (OPG) and the soluble form of the receptor activator for nuclear factor (NF)-kappaB (RANK-L), also are candidate markers of the bone metastases process and offer potential as surrogate markers of tumor-induced osteoclastogenesis. METHODS: Immunoassays have been developed that can determine blood and urine levels of the telopeptides associated with bone collagen breakdown. In addition, immunoassays now are available that can measure both OPG and the soluble form of RANK-L in blood and tissues as markers of osteoclastogenesis. RESULTS: The measurement of bone collagen breakdown products and specific factors associated with the process of osteoclastogenesis have been applied to the management and assessment of patients with metastatic bone disease. Results indicate that these surrogate markers can be useful in determining response to antiresorptive therapy, in selecting optimal dosage, and as markers of disease progression. CONCLUSIONS: Although they show promise, questions remain whether blood measurements of surrogate markers of osteoclastogenesis, like OPG and RANK-L, will be useful in both assessing and managing patients with metastatic bone disease. PMID- 12548590 TI - Current and future status of adjuvant therapy for breast cancer. AB - Adjuvant systemic treatments have greatly improved the prognosis of women with early breast cancer. Combination chemotherapy and, for patients with oestrogen receptor-positive (ER+) tumours, endocrine treatment has been found to reduce the frequency of relapse and improve survival. New adjuvant strategies include the introduction of taxanes into adjuvant chemotherapy schedules, the use of aromatase inhibitors in place of, or in addition to, tamoxifen, and the use of adjuvant bisphosphonates. Combination chemotherapy has been found to reduce the annual odds of recurrence and death in pre- and postmenopausal women. The benefits, however, are on average less in older patients. Anthracycline-based regimens are more effective than traditional regimens of cyclophosphamide, methotrexate, and fluorouracil (CMF). The benefits of adjuvant cytotoxic and endocrine treatments are additive. There is considerable debate as to the role of taxanes in adjuvant therapy. Improved outcome has been observed in one large trial, especially in those patients with ER-negative tumours. High-dose chemotherapy has not fulfilled its early promise. Ovarian suppression and/or tamoxifen remain the treatments of choice. The annual odds of relapse and death have been reduced by approximately one-third and one-quarter, respectively. Several very large studies are in progress to assess the potential of aromatase inhibitors in the adjuvant setting. Direct comparisons with tamoxifen, as well as switching after several years from tamoxifen to an aromatase inhibitor, are strategies under evaluation. Early results from one of these trials evaluating anastrozole (the Arimidex, Tamoxifen, Alone or in Combination [ATAC] trial) has reported a reduced relapse rate after a median follow-up of 3 years in favour of anastrozole. However, this was at the expense of accelerated bone loss, and strategies to minimise this side effect of aromatase inhibitors are under investigation. Although many studies have indicated that bisphosphonates prevent the development of metastatic bone disease in animals, the clinical role of prophylactic bisphosphonates in early breast cancer is not clearly defined. Three studies with oral clodronate have been published, two of them indicating a protective effect on the development of bone metastases and improved survival, and one suggesting a disadvantage to the use of adjuvant clodronate. Further large adjuvant trials with clodronate and zoledronic acid are in progress. Adjuvant bisphosphonates also have been found to reduce bone loss associated with cancer treatments and preserve skeletal health. It may be possible to replace the current oral regimens for prevention of bone loss with a single annual infusion of the highly potent bisphosphonate zoledronic acid. PMID- 12548591 TI - A phase I study of AMGN-0007, a recombinant osteoprotegerin construct, in patients with multiple myeloma or breast carcinoma related bone metastases. AB - BACKGROUND: Osteoprotegerin (OPG) is a decoy receptor for OPG ligand (OPGL), or receptor activator of NF-kappaB ligand (RANKL). RANKL/RANK interaction is important in terminal differentiation and activation of osteoclasts. In binding to RANKL, OPG blocks differentiation and activation of osteoclasts. AMGN-0007 is a recombinant OPG construct developed as a potential therapeutic agent in the treatment of bone disease. METHODS: A randomized, double-blind, double-dummy, active-controlled, single-dose, dose escalation study was conducted to determine the safety and effect on bone resorption of AMGN-0007 in patients with multiple myeloma (n = 28) or breast carcinoma (n = 26) with radiologically confirmed lytic bone lesions. Patients were randomized (3:1 ratio) to receive a single dose of either AMGN-0007 (subcutaneously [SC]) or pamidronate (90 mg intravenously) and were followed for 56 days. Medications or other diseases affecting bone metabolism and chemotherapy within 28 days of dosing were exclusion criteria. Biologic activity of AMGN-0007 was assessed by measurement of the surrogate marker of bone resorption, urinary N-telopeptide of collagen (NTX). RESULTS: AMGN 0007 caused a rapid, sustained, dose-dependent decrease in NTX/creatinine levels, which was at least comparable to the profile observed with pamidronate. Four serious adverse events were reported, three in breast carcinoma patients: a fracture in the left femur (pamidronate, considered unrelated), extreme fatigue (0.3 mg/kg AMGN-0007, considered unrelated), and congestive heart failure (1.0 mg/kg AMGN-0007, considered by the investigator to be probably related to doxorubicin and radiation therapy); one event occurred in a multiple myeloma patient: Herpes zoster (pamidronate, considered unrelated). Two multiple myeloma patients (1.0 mg/kg AMGN-0007) had albumin-adjusted serum calcium levels of 1.9 mmol/L on Day 8 but without clinical symptoms. CONCLUSIONS: A single SC dose of AMGN-0007 suppressed bone resorption as indicated by a rapid, sustained, and profound decrease of urinary NTX/creatinine in multiple myeloma and breast carcinoma patients. Changes were comparable to those with pamidronate. AMGN-0007 was well tolerated. PMID- 12548593 TI - Impact of germline BRCA1 mutations and overexpression of p53 on prognosis and response to treatment following breast carcinoma: 10-year follow up data. AB - BACKGROUND: Overexpression of p53 has been associated with poor survival following breast carcinoma. BRCA1 interacts biochemically with p53 and may also contribute to poor outcome when constitutionally mutated. The joint effect of both abnormalities has not been studied. The primary objective of this study was to assess the impact of germline BRCA1 mutations and p53 overexpression on survival after 10 years of follow-up. METHODS: A historical cohort of Ashkenazi Jewish women 65 years or younger with invasive breast carcinoma was tested for BRCA1 founder mutations. p53 overexpression was assessed by immunohistochemistry. Clinicopathologic information was obtained by chart review. RESULTS: In total, 278 women were analyzed. On univariate analysis, p53 overexpression (n = 63) was prognostic for worse overall survival (relative risk [RR] 2.6, P = 0.001) whereas BRCA1 germline mutations (n = 30) were of borderline significance (RR 1.9, P = 0.052). In the lymph node-negative subpopulation, BRCA1 mutation status conferred a higher mortality on univariate (RR 5.6, P < 0.001) and multivariate (RR 3.5, P = 0.03) analyses. There was a trend in favor of a worse prognosis for women who carried a germline BRCA1 mutation and whose tumor overexpressed p53. When compared with noncarriers, BRCA1 mutation carriers had a worse overall survival if they did not receive adjuvant chemotherapy (RR 3.3, P= 0.01) or adjuvant hormonal therapy (RR 2.3, P = 0.02). CONCLUSIONS: Germline BRCA1 mutations and p53 overexpression carry a negative prognosis that is not additive to known prognostic factors. Given the experimental sensitivity of BRCA1-mutated cells to chemotherapy, the worse survival among BRCA1 mutation-carrying lymph node negative breast carcinoma patients may be partly explained by the significantly lower proportion of lymph node-negative patients who received adjuvant chemotherapy (P < 0.001). PMID- 12548594 TI - A phase II trial of docetaxel and estramustine in patients with refractory metastatic breast carcinoma. AB - BACKGROUND: The similarity between the mechanism of action between docetaxel and estramustine generated the hypothesis of synergistic antimicrotubule effects and cytotoxicity when the two agents are combined. In addition, it has been demonstrated that estramustine binds P-glycoprotein in vitro and, thus, may prevent the efflux of taxanes in tumors that over-express P-glycoprotein. To further evaluate the combinations clinical efficacy and safety, a trial was performed in heavily pretreated patients with metastatic breast carcinoma (MBC). METHODS: Thirty-six patients with MBC were treated with estramustine 900 mg/m(2) per day divided into 3 doses given on Days 1-3 and docetaxel 70 mg/m(2) given by intravenous administration over 1 hour on Day 3 after the first dose of estramustine, every 21 days. Patients may have received any number of prior chemotherapy regimens for MBC. RESULTS: Nine partial responses were observed in 31 assessable patients, for an objective response rate of 29% (95% confidence interval, 14-48%). The median progression free survival was 4 months (range, 1-41 months), and the median overall survival was 17 months (range, 2-45 months). Severe toxicities (Grade 3 or 4) were neutropenia, hypophosphatemia, and thrombosis. Seventy-five percent of patients experienced either an improvement or no change in quality of life. CONCLUSIONS: The combination of docetaxel and estramustine produced responses in heavily pretreated women with MBC while maintaining quality of life. PMID- 12548595 TI - Survival of patients with metastatic breast carcinoma: importance of prognostic markers of the primary tumor. AB - BACKGROUND: Women with metastatic breast carcinoma have a highly variable clinical course and outcome. Intrinsic genetic heterogeneity of the primary breast tumor may play a role in this variability and may explain it in part. Therefore, the authors tested the hypothesis that the characteristics of primary breast tumors are important determinants of prognosis and survival in patients with metastatic breast carcinoma. METHODS: The prognostic significance of the biology of the primary tumor for outcome in patients with metastatic breast disease was assessed in 346 patients with lymph node positive breast carcinoma who developed distant, recurrent disease. Traditional prognostic indicators (age, tumor size, number of involved lymph nodes, sites of recurrence, disease free interval [DFI], adjuvant treatments, estrogen receptor [ER] expression, progesterone receptor [PgR] expression, S-phase fraction [SPF], and DNA ploidy), together with three newer biologic markers (c-erbB-2, p53, and bcl-2) were assessed. Sites of recurrence were defined as nonvisceral (bone and locoregional lymph nodes) or visceral (lung, liver, brain, and other organs). RESULTS: The median duration of survival was 17.8 months (95% confidence interval, 15.2-21.5 months). Univariate analysis showed that age > 50 years, visceral disease, and shorter DFI were associated significantly with poor outcome (P < 0.05). In addition, the molecular phenotype of the primary breast tumor was significant, with primary tumors that showed ER negativity and PgR negativity, high SPF, aneuploidy, accumulation of p53 protein, and lower bcl-2 expression, together with c-erbB-2 overexpression, all associated with a poorer clinical outcome (P < 0.05). In a multivariate analysis, older age, visceral disease, shorter DFI, PgR negativity, high SPF, and lower bcl-2 expression were significant predictors of worse survival (P < 0.05). CONCLUSIONS: In addition to traditional risk factors, bcl-2 negativity was associated significantly with a worse clinical outcome. Biologic features of primary tumors were correlated independently with outcome after first recurrence in patients with metastatic breast carcinoma and may be used as indicators of prognosis in the metastatic setting. PMID- 12548596 TI - Radiofrequency ablation of adrenal tumors and adrenocortical carcinoma metastases. AB - BACKGROUND: The current study was performed to analyze the feasibility, safety, imaging appearance, and short-term efficacy of image-guided percutaneous radiofrequency ablation (RFA) of primary and metastatic adrenal neoplasms including adrenocortical carcinoma. METHODS: The procedure was performed using 36 treatment spheres on 15 adrenocortical carcinoma primary or metastatic tumors in eight patients over 27 months. Tumors ranged from 15 to 90 mm in greatest dimension with a mean of 43 mm. All patients had unresectable tumors or were poor candidates for surgery. Mean follow-up was 10.3 months. RESULTS: All patients were discharged or were free of procedure-related medical care 6-48 hours after the procedures without major complications. All treatments resulted in presumptive coagulation necrosis by imaging criteria, which manifested as loss of previous contrast enhancement in ablated tissue. Eight of 15 (53%) posttreatment thermal lesions lost enhancement and stopped growing on latest follow-up computed tomographic scan. Three of 15 (20%) demonstrated interval growth and four did not change in size. Of these four lesions, two showed contrast enhancement. For smaller tumors with a mean greatest dimension less than or equal to 5 cm, 8 of 12 (67%) tumors were completely ablated, as defined by decreasing size and complete loss of contrast enhancement. Three of 15 (20 %) tumors and related thermal lesions were found to have disappeared nearly completely on imaging. CONCLUSIONS: Percutaneous, image-guided RFA is a safe and well tolerated procedure for the treatment of unresectable primary or metastatic adrenocortical carcinoma. The procedure is effective for the short-term local control of small adrenal tumors, and is most effective for tumors less than 5 cm. The survival rate for patients with adrenocortical carcinoma improves when radical excision is performed in selected patients. Aggressive local disease control may potentially influence survival as well. However, further study is required to evaluate survival impact, document long-term efficacy, and to determine if RFA can obviate repeated surgical intervention in specific clinical scenarios. PMID- 12548597 TI - Phase II trial of cyclophosphamide, vincristine, and dexamethasone in the treatment of androgen-independent prostate carcinoma. AB - BACKGROUND: In this Phase II study, the authors assessed the toxicity and anti tumor activity of a combination of oral cyclophosphamide, oral low-dose dexamethasone, and intravenous vincristine (CVD) in patients with metastatic androgen-independent prostate carcinoma (AI-PCa). METHODS: Patients with histologic proof of adenocarcinoma of the prostate progressing despite adequate hormonal therapy and adequate organ function were treated with oral cyclophosphamide, 250 mg/daily (Days 1-14); intravenous vincristine, 1 mg daily (Days 1, 8, 15); and oral dexamethasone, 0.75 mg twice a day (Days 1-14) in 28 day cycles. Study endpoints were toxicity, rate of prostate specific antigen (PSA) decline > 50%, and/or measurable disease response. RESULTS: Fifty-two (95%) of 55 registered patients were evaluable. The majority (65%) of patients had received prior chemotherapy. The median number of treatment cycles given was two (range, one-seven cycles). Twenty-nine percent of the patients were found to have a > 50% decline in PSA level compared with baseline levels, and 25% of the patients with bidimensionally measurable soft-tissue or visceral disease were found to have a partial response. The median progression-free survival duration was 10 weeks, and the median overall survival duration was 10.6 months. There were no thromboembolic events, and hematologic and nonhematologic toxicity was minimal. CONCLUSIONS: CVD was found to be an active and well-tolerated regimen for AI-PCa. The low toxicity profile makes CVD a useful treatment option for patients with significant comorbidities and high risk for treatment-related toxicity, especially thromboembolic events and myelotoxicity. PMID- 12548598 TI - Prognostic factors in neuroendocrine small cell cervical carcinoma: a multivariate analysis. AB - BACKGROUND: The purpose of this study was to evaluate the clinical and pathologic factors associated with survival in patients with neuroendocrine (NE) cervical carcinoma. METHODS: All patients with NE cervical carcinoma diagnosed between 1979-2001 were identified from tumor registry databases at two hospitals. Data were collected from hospital charts, office records, and tumor registry files. The impact of clinical and pathologic risk factors on the survival of patients with small cell NE carcinoma of the cervix was evaluated using Kaplan-Meier life table analyses and log-rank tests. The independent prognostic factors found to be predictive of survival in univariate analysis were evaluated using Cox regression. All tests were two-tailed with P values < 0.05 considered significant. RESULTS: Thirty-four patients (median age, 42 years) were diagnosed with neuroendocrine cervical carcinoma, which included 21 with International Federation of Gynecology and Obstetrics (FIGO) Stage I disease, 6 with FIGO Stage II disease, 5 with FIGO Stage III disease, and 2 with FIGO Stage IV disease. Seventeen patients underwent a radical and 6 patients underwent a simple hysterectomy. Fourteen women received adjuvant therapy with pelvic radiation and/or cisplatin-based chemotherapy. Ten women received primary radiotherapy with (n = 5) or without (n = 4) chemotherapy and the remaining patient refused therapy. Women with early-stage (Stage I-IIA) disease had median survival rates of 31 months compared with 10 months in the advanced-stage (Stage IIB-IVB) group (P = 0.002). In univariate analysis, advanced stage (P = 0.002), tumor size >2 cm (P = 0.02), margin involvement (P = 0.016), pure versus a mixed histologic pattern (P = 0.04), margin status (P = 0.016), and smoking (P = 0.04) were considered poor prognostic factors. In multivariate analysis, smoking for early stage patients and stage of disease in the overall population remained as independent prognostic factors of survival. CONCLUSIONS: Smoking and advanced stage are reported to be poor prognostic factors for survival in patients with NE small cell carcinoma of the cervix. Only those with early lesions amenable to extirpation are cured. The role of primary or postoperative radiation with or without chemotherapy is unclear and yields uniformly poor results, particularly in patients with advanced lesions. Clinical trials are needed. PMID- 12548599 TI - R-Ras promotes tumor growth of cervical epithelial cells. AB - BACKGROUND: R-Ras is 55% identical to H-Ras. However, these two oncogenes seem to have different tumor-transforming potential. R-Ras induced cell transformation in fibroblasts but not in other cell types. R-Ras also reportedly induces a more invasive phenotype in breast epithelial cells through integrin activation. The authors studied the mechanisms whereby R-Ras induces a malignant phenotype. METHODS: Dominant negative (R-Ras43N) and constitutively active (R-Ras87L) mutants of R-Ras were stably transfected into human cervical epithelium C33A cells. Transfected cells were analyzed for adhesion, cell spreading, migration, and growth in culture and in nude mice. The activity of extracellular signal regulated kinase (ERK) and phosphatidylinositol 3-kinase (PI 3-K) also was determined by Western blot analysis and by in vitro kinase assays. RESULTS: R Ras87L-transfected cells, but not R-Ras43 N-transfected cells, had a higher growth rate in nude mice and in culture compared with control cells. None of the transfected C33A cells showed an increase in cell adhesion to fibronectin or collagen I, nor did they show an increment of beta1 integrin affinity. However, cells that expressed R-Ras87L, but not cells that expressed R-Ras 43N, presented a marked increase in cell spreading and migration through collagen-coated membranes. Increases in cell proliferation, spreading, and migration induced by R Ras87L were inhibited by the PI 3-K inhibitor LY294002. In addition, PI 3-K activity, but not ERK activity, was increased only in cells that expressed R Ras87L. CONCLUSIONS: These data suggest that the oncogene R-Ras promotes tumor growth of cervical epithelial cells and increases their migration potential over collagen through a pathway that involves PI 3-K. PMID- 12548600 TI - Frequency of gastrointestinal involvement and its clinical significance in mantle cell lymphoma. AB - BACKGROUND: The reported frequency of gastrointestinal (GI) tract involvement in patients with mantle cell lymphoma (MCL) is 15-30%. However, this figure most likely is an underestimate because most patients with MCL involving the GI tract previously reported were examined endoscopically only if they had GI tract symptoms. The impact of endoscopic assessment on the management of MCL patients is unknown. METHODS: From March 1998 to May 2001 baseline upper and lower endoscopy of the GI tract was performed in consecutive untreated patients with MCL as part of a prospective therapeutic trial. Biopsies were performed on abnormal as well as macroscopically normal mucosa. Endoscopy was repeated during treatment and as part of follow-up evaluations. RESULTS: Only 26% of patients presented with GI symptoms at the time of diagnosis. MCL was present histologically in the lower GI tract of 53 of 60 patient (88%) and in the upper GI tract of 28 of 58 patients (43%). Microscopic evidence of MCL was found in 84% of patients with normal visual (macroscopic) findings by lower endoscopy and in 45% of patients with macroscopically normal findings by upper endoscopy. Despite this high frequency of GI tract involvement, the use of upper and lower endoscopy with biopsies in this group of patients resulted in changes in clinical management in only three (4%) patients. CONCLUSIONS: Gastrointestinal tract involvement was found to be present in most patients with MCL, usually at a microscopic level involving macroscopically normal mucosa. The use of aggressive staging evaluation of the GI tract was found to have little impact on patient management decisions in the current study. PMID- 12548601 TI - Treatment options for patients with acute myeloid leukemia with a matched sibling donor: a decision analysis. AB - BACKGROUND: The role of allogeneic bone marrow transplantation (BMT) in the consolidation of young adults with acute myeloid leukemia (AML) with matched sibling donors (MSD) is controversial. Although BMT is associated with increased event free survival compared with intensive chemotherapy (CT) consolidation, BMT also is associated with increased treatment-related mortality and likely decreased quality of life and life expectancy in patients who do not develop recurrent disease. METHODS: The authors used decision analysis to compare three strategies for maximizing quality-adjusted life years (QALYs) in patients with AML in first remission with an MSD: BMT All, BMT None (consolidation CT only), or BMT in high-risk patients, as defined by baseline cytogenetic testing (Test strategy). A second decision-analysis tree was then constructed that compared BMT with CT specifically for patients with intermediate cytogenetics. RESULTS: Using expected QALYs as the outcome measure, the Test, BMT All, and BMT None strategies were associated with 20.10 QALYs, 19.63 QALYs, and 18.38 QALYs, respectively. Thus, the Test strategy, with CT for low-risk patients and BMT for intermediate risk and high-risk patients, was expected to be the optimal strategy. In the intermediate cytogenetic decision analysis, although the expected QALY for BMT recipients was higher compared with CT recipients (19.78 QALYs vs. 18.75 QALYs), because of uncertainty in variable estimates, the optimal choice was less clear. CONCLUSIONS: CT consolidation is a reasonable option for patients with AML who have favorable cytogenetics, even if an MSD is available. This model provides a framework from which patients with AML and their physicians can make decisions about consolidation therapy. PMID- 12548602 TI - Fluorescence in situ hybridization analysis of aneuploidization patterns in monoclonal gammopathy of undetermined significance versus multiple myeloma and plasma cell leukemia. AB - BACKGROUND: Monoclonal gammopathy of undetermined significance (MGUS) is a clonal plasma cell (PC) disorder usually characterized by a benign clinical course. However, in approximately 25% of patients, the disorder has been found to evolve into a multiple myeloma (MM). The mechanism leading to the evolution of MGUS remains unknown. The aim of the current study was, first, to assess by interphase fluorescence in situ hybridization (FISH) the incidence of numerical abnormalities of chromosomes 6, 9, 13, and 17 in MGUS patients and to compare it with that found in MM and PC leukemia (PCL) patients and, second, to explore the potential heterogeneity of the pathologic PC in MGUS as a way to identify unique cytogenetic patterns different from those frequently observed in MM and PCL. METHODS: Numerical abnormalities of chromosomes 6, 9, 13, and 17 were investigated by dual- and triple-color FISH in bone marrow PC from 208 patients corresponding to MGUS (n = 30), MM (n = 158), and PCL (n = 20) cases. In MGUS and MM patients with < 10% PC, both normal and phenotypically aberrant PC were discriminated by multiparameter flow cytometry, the latter subset being specifically sorted for FISH analysis with a purity of 93% +/- 6%. RESULTS: Overall, 57% of the MGUS patients displayed abnormalities for at least 1 of the 4 chromosomes analyzed compared with 75% of both MM and PCL cases. The most common single chromosome abnormalities detected in MGUS were gains of chromosomes 9 (23%) and/or 6 (21%) and loss of chromosomes 13 (21%) and/or 17 (17%). Compared with MM patients, MGUS patients were found to have both a lower incidence of gains of chromosome 9 (23% vs. 54%, P = 0.002) and monosomy 13/13q(-) deletions (21% vs. 38%, P = 0.07); with respect to PCL cases, MGUS patients were found to have a lower incidence of monosomy 13/13q(-) deletions (21% vs. 75%, P < 0.001) together with a slightly higher frequency of gains of both chromosomes 6 (21% vs. 0%, P = 0.05) and 9 (23% vs. 7%, P = 0.1). The simultaneous use of two or three different chromosome probes showed that within the purified compartment of phenotypically aberrant PC from most MGUS patients (67%), more than 1 PC clone could be identified. In contrast, the incidence of 2 or more PC clones was much lower in MM (19%, P < 0.001) and PCL (15%, P = 0.003). Interestingly, although some FISH patterns were shared by both groups of diseases (i.e., monosomy 13/13q( ) deletions alone, gains of chromosome 9 alone or together with trisomy 6), others were found almost exclusively in either MGUS (i.e., a clone with monosomy 6 and/or 17 together with nuclei displaying a normal chromosome number) or in MM (i.e., monosomy 13/13q(-) deletions together with gains of chromosome 6 and/or 9). CONCLUSIONS: In summary, the results of the current study showed that MGUS patients displayed a high incidence of numerical alterations, which are usually associated with the presence of more than one tumor cell clone. It is interesting to note that the cytogenetic patterns observed in the aneuploid PC clones from MGUS patients were frequently different from those observed in both MM and PCL. PMID- 12548604 TI - Moderate sun exposure and nevus counts in parents are associated with development of melanocytic nevi in childhood: a risk factor study in 1,812 kindergarten children. AB - BACKGROUND: Melanocytic nevi have been identified as the most important risk factor for cutaneous melanoma. Sun exposure, sunburns, and light pigmentation have been found to be associated with their development in childhood. To the authors' knowledge, nevus proneness of parents and the exact type of ultraviolet (UV) exposure have not yet been investigated in this context. The authors' objective was to determine independent risk factors and their impact for nevus development in childhood. METHODS: The current study was conducted by two university departments of dermatology in 49 public nursery schools in Stuttgart, Germany and in 38 public nursery schools in Bochum, Germany. The cross-sectional study included 1,812 children aged 2-7 years and their parents. Total body nevus counts in children, assessment of pigmentary features, and nevus counts on the arms of parents were performed. Parents underwent a standardized interview concerning national origin and lifestyle features, as well as habits and magnitude of sun exposure of children. Analysis was performed by multivariate linear regression analysis and by multiple logistic regression analysis. RESULTS: The number of nevi was found to steadily increase with age from a median of 3 at age 2 years to 19 at age 7 years (P < 0.0001). High numbers of nevi in children were associated with the number of weeks on sunny holidays, outdoor activities at home, skin type, facial freckling, ethnicity of parents, and the number of nevi on the arms of parents. Previously experienced sunburns failed significance (P = 0.0620). CONCLUSIONS: The authors found a strong association between nevus development in children and the number of parental moles, which most likely points to an inherited factor. Moderate sun exposure such as outdoor activities during a German summer without sunburns seemed to be sufficient for induction of melanocytic nevi. The authors believe that these findings will have direct impact on concepts for preventive strategies. PMID- 12548603 TI - Cytotoxic/natural killer cell cutaneous lymphomas. Report of EORTC Cutaneous Lymphoma Task Force Workshop. AB - BACKGROUND: Cutaneous lymphomas expressing a cytotoxic or natural killer (NK) cell phenotype represent a group of lymphoproliferative disorders for which there is currently much confusion and little consensus regarding the best nomenclature and classification. METHODS: This study analyzes 48 cases of primary cutaneous lymphoma expressing cytotoxic proteins and/or the NK cell marker, CD56. These cases were collected for a workshop of the European Organization for Research and Treatment of Cancer Cutaneous Lymphoma Task Force, to better clarify the clinical, morphologic, and phenotypic features of these uncommon tumors. RESULTS: Several categories with different clinical and pathologic features were delineated: 1) aggressive, CD8+, epidermotropic, cytotoxic T-cell lymphoma; 2) mycosis fungoides, cytotoxic immunophenotype variant; 3) subcutaneous panniculitis-like T-cell lymphoma; 4) NK/T-cell lymphoma, nasal type; 5) CD4+, NK cell lymphoma; 6) blastoid NK cell lymphoma; (7) intravascular NK-like lymphoma; and 8) cytotoxic, peripheral T-cell lymphoma. CONCLUSIONS: Our data show that primary cutaneous cytotoxic/NK cell lymphomas include distinct groups of diseases, clinically, histologically, and biologically. Because the finding of a cytotoxic phenotype often has prognostic significance, the routine use of cytotoxic markers in the diagnosis and classification of cutaneous lymphomas should be expanded. PMID- 12548605 TI - A population-based analysis of risk factors for a second primary cutaneous melanoma among melanoma survivors. AB - BACKGROUND: The results of several studies have provided evidence that patients diagnosed with cutaneous melanoma (CM) are at a higher risk of developing a second primary CM than the general population. In this study, the authors examined how the risk of a second primary tumor varied with time from diagnosis of CM and examined the patient-specific factors that modify a CM patient's risk of developing a second primary tumor. METHODS: Survival curves for time from first CM to second CM were calculated using the Kaplan-Meier method. The Cox proportional hazards model was used to determine which demographic- and disease related factors influence the risk of a second CM. RESULTS: Approximately 0.5% of Surveillance, Epidemiology, and End Results (SEER) CM patients were found to have synchronous second primaries. The estimated cumulative probability of having a second primary CM was 0.99% at 1 year after initial CM diagnosis, 2.06% at 5 years, 3.17% at 10 years, and 5.34% at 20 years. Risk was significantly greater for males; older patients; patients with first CM on the face, neck, or trunk; those from the Atlanta, Hawaii, or Connecticut registries; and more recently diagnosed patients. Risk was lower for patients from the Utah registry and those with Stage IV disease. CONCLUSIONS: The elevated risk for CM among CM survivors appears to be greatest in the first few months, and then subsequently declines. However, the risk for a second CM among CM survivors was found to remain substantially higher than the risk for a first CM in the general population throughout the observation period (> 20 years). Demographic- and disease-related factors substantially modify the risk of a second primary CM. PMID- 12548606 TI - Low expression of p27 indicates a poor prognosis in patients with high-grade astrocytomas. AB - BACKGROUND: Two families of tumor suppressor genes, Cip/Kip (p21, p27, and 57) and INK4 (p15, p16, p18, and p19), regulate cell proliferation and neoplastic transformation. p27 exerts its suppressor effect through cyclin E-dependent kinase (CDK2) by inhibiting the phosphorylation of pRb by CDK2, which, in turn, arrests cells in the G1-phase. p21 has a similar effect in addition to participating in the p53 dependent CDK4-mediated and CDK6-mediated pathway. The authors studied the prognostic significance of p21 and p27 in patients with high grade astrocytomas who were treated with radiotherapy. METHODS: The expression of p27 and p21 was analyzed immunohistochemically in 52 glioblastomas and 25 anaplastic astrocytomas. All patients underwent surgery for the first time and were treated with adjuvant external radiotherapy. RESULTS: The p27 labeling index (LI) was < 30% in 36% of tumors, 30-50% in 25% of tumors, and > 50% in 39% of tumors. A significant difference in cumulative survival was observed between these groups (P = 0.0072; log-rank test). The p21 LI was < 30% in 48% of tumors, 30-50% in 39% of tumors, and > 50% in 13% of tumors; these groups did not differ significantly in survival. In multivariate Cox analysis, p27 LI was an independent prognostic factor (P = 0.0008). The grade of malignancy and proliferation activity also were independent prognostic factors. CONCLUSIONS: Although p27 and p21 are parallel cell-cycle regulators, only p27 has independent prognostic value in patients with malignant astrocytomas. It appears that decreased levels of p21/p27 are associated with a poor prognosis and short survival. PMID- 12548607 TI - The predictive value of longitudinal neuropsychologic assessment in the early detection of brain tumor recurrence. AB - BACKGROUND: Neuropsychologic tests are widely used to predict the course of progressive neurologic diseases, and recent research has demonstrated the specificity of cognitive measures, even in relatively diffuse diseases. However, the cognitive effects of brain tumors of similar histology and location are known to be highly variable. The authors used the specificity of cognitive function principle to compare two models for the early detection of low-grade brain tumor recurrence prior to detection with clinically scheduled neuroimaging. METHODS: To test the feasibility of these prediction models, 34 patients with supratentorial, low-grade brain tumors prospectively were administered serial comprehensive neuropsychologic examinations; 11 patients developed recurrent tumors during the series and 23 patients did not. A general model based on tests sensitive to malignancy and white matter disease was compared with a tumor-specific model based on indices related to each patient's tumor locus. A Cox proportional hazards model was used to identify the predictor variables that significantly changed immediately prior to recurrence. RESULTS: Only the tumor-specific model achieved statistical significance (P < 0.02). A tumor-specific index decline of 1 standard deviation indicated a 5-fold increase in the probability of tumor recurrence. CONCLUSIONS: Although this method needs to be tested with more frequent and regular observations and with a larger sample, these results provide evidence of the feasibility of the subject-specific model as a predictor of recurrence. The evidence of the predictive value of a tumor-specific model is consistent with studies that identify only limited, brain structure-specific cognitive decline from broad neuropsychologic batteries. PMID- 12548608 TI - A prospective study on glioblastoma in the elderly. AB - BACKGROUND: Elderly patients (age > 65 years) with glioblastoma multiforme frequently are excluded from clinical studies, and prospective trials for patients with this age group do not exist to date. METHODS: The authors conducted a prospective trial in 79 consecutive elderly patients with glioblastoma who underwent surgery and received radiotherapy (59.44 grays in 33 fractions; Group A; n = 24 patients) or received the same radiotherapy plus adjuvant chemotherapy with procarbizine, lomustine, and vincristine (PCV; lomustine 110 mg/m(2) on Day 1, procarbazine 60 mg/m(2) on Days 8-21, and vincristine 1.4 mg/m(2) on Days 8 and 29 every 42 days; Group B; n = 32 patients), or received the same radiotherapy plus adjuvant temozolomide (150 mg/m(2) for 5 days every 28 days; Group C; n = 22 patients). RESULTS: The median time to disease progression (TTP) and median survival MST were 7.2 months (95% confidence interval [95%CI], 6.34 8.64) and 12.5 months (95%CI, 11.6-14.8), respectively. The TTP was significantly better for Group C compared with Groups A and B (10.7 months vs. 5.3 months and 6.9 months, respectively; P = 0.0002). Karnofsky performance status (KPS) (P < 0.001) and temozolomide (P < 0.001) were the only independent prognostic factors. Overall survival was better in Group C compared with Group A (14.9 months vs. 11.2 months; P = 0.002), but there were no statistical differences found between Groups A and B or between Groups B and C. Only KPS (P < 0.001) was predictive of overall survival, even if temozolomide chemotherapy was very close to the significance level (P = 0.058). Hematologic Grade 3-4 toxicity was higher with the PCV chemotherapy regimen compared with the temozolomide chemotherapy regimen. CONCLUSIONS: Age alone should not preclude appropriate treatment in elderly patients with good performance status, for whom definitive radiation therapy and adjuvant chemotherapy with temozolomide is advised. PMID- 12548609 TI - Endocrine and cardiovascular late effects among adult survivors of childhood brain tumors: Childhood Cancer Survivor Study. AB - BACKGROUND: Survivors of childhood brain tumors (CBTs) are at high risk for a variety of late adverse effects. Most research on long-term effects of CBTs has been comprised of single-institution case series without comparison groups. Research on CBT late effects often is focused on neurologic and sensory outcomes, with less emphasis on other potential targets such as the endocrine and circulatory systems. The current study was conducted to contrast the incidence of endocrine and cardiovascular conditions among CBT survivors as a function of treatment and to determine the risk of occurrence of these conditions relative to a sibling comparison group. METHODS: As part of the Childhood Cancer Survivor Study (CCSS), treatment data were collected from medical records and self reported late effects were ascertained from a survey questionnaire of 1,607 CBT patients who survived their disease for 5 or more years. For comparison purposes, questionnaire data were also collected from 3418 randomly selected siblings of participants in CCSS. RESULTS: One or more endocrine conditions were reported by 43% of CBT survivors. Compared with siblings, CBT survivors had a significantly increased risk of late-onset (>/= 5 years postdiagnosis) hypothyroidism (relative risk [RR] = 14.3; 95% confidence interval [95% CI] 9.7-21.0), growth hormone deficiency (RR = 277.8; 95% CI 111.1-694.9), the need for medications to induce puberty (RR = 86.1; 95% CI 31.1-238.2), and osteoporosis (RR = 24.7; 95% CI 9.9 61.4). One or more cardiovascular conditions were reported by 18% of CBT survivors, with an elevated late-onset risk for stroke (RR = 42.8; 95% CI 16.7 109.8), blood clots (RR = 5.7; 95% CI 3.2-10.0), and angina-like symptoms (RR = 2.0; 95% CI 1.5-2.7). Very few late effects were evident among those treated with surgery only, but risks were consistently elevated for those treated with radiation and surgery, and higher still for those who also received adjuvant chemotherapy. CONCLUSIONS: Childhood brain tumor survivors are at a significantly increased risk for several adverse endocrine and cardiovascular late effects, particularly if they were treated with radiation and chemotherapy. Lifetime medical surveillance and follow-up for potential toxicities are necessary because treatment-related complications may occur many years after therapy. PMID- 12548610 TI - Health-related quality of life of cancer and noncancer patients in Medicare managed care. AB - BACKGROUND: Data from the Health Care Financing Administration's (HCFA) Medicare Health Outcomes Survey (MHOS) of patients enrolled in managed care services through Medicare were analyzed. The MHOS provided baseline estimates of quality of life of cancer survivors in comparison to a frequency age-matched cohort of noncancer patients. METHOD: In 1998, the MHOS was mailed to a random sample of 279,135 beneficiaries. Completed surveys were received from 167,096 respondents (60%). Some 22,747 respondents who had been diagnosed with cancer were frequency age matched to an equal number of noncancer patients. RESULTS: Cancer survivors had statistically significantly poorer scores than noncancer patients on all eight subscales as well as on the Physical Component and Mental Component summary measures of the Medical Outcomes Study Short Form-36 (MOS SF-36). Comparisons by type and number of cancers for which an individual was currently in treatment showed lowest quality of life for those in treatment for lung carcinoma, followed by those who were in treatment for more than one type of cancer. CONCLUSION: The data suggest that cancer shows negative effects on health-related quality of life that are not explainable by simple effects of age because frequency age-matched cancer survivors had statistically significantly lower scores on all 10 scores of the MOS SF-36. Effect sizes are evaluated to determine the clinical significance of these differences in health-related quality of life. The MHOS offers useful data for planning and improving cancer policy and programs. PMID- 12548611 TI - Titration of serum p53 antibodies in 1,085 patients with various types of malignant tumors: a multiinstitutional analysis by the Japan p53 Antibody Research Group. AB - BACKGROUND: There have been very few large-scale, multiinstitutional studies of surveillance of serum p53 antibodies (S-p53 Abs) in patients with various malignant tumors. METHODS: A highly specific, quantitative enzyme-linked immunosorbent assay (ELISA) kit was developed and used to evaluate the efficiency of detecting p53 Abs. A cut-off value was established by analyzing sera from 205 healthy volunteers as reference individuals. Sera from 1085 patients with various types of primary malignant tumors were studied for the presence of S-p53 Abs before treatment. Sera from 34 patients were selected randomly for a competition assay to ensure that antibodies were specific to p53 protein. Carcinoembryonic antigen (CEA) was assessed to compare its positive rate with the positive rate of S-p53 Abs. RESULTS: The median value of S-p53 Abs in healthy control individuals was 0.33 U/mL (range, 0.0-4.39 U/mL). Based on reference values that were calculated using parametric determination of the lower 0.95 fraction of the reference distribution in healthy control individuals, the cut-off value was determined as 1.3 U/mL. Two hundred twenty-one of 1085 patients (20.4%) were positive for S-p53 Abs. The highest relevance of S-53 Abs was associated with head and neck carcinoma (32%), followed by esophageal carcinoma (30%), colorectal carcinoma (24%), and carcinoma of the uterus (23%). The positive rate for S-p53 Abs was higher compared with the positive rate for CEA in patients with squamous cell carcinoma. CONCLUSIONS: Surveillance of S-p53 Abs is useful in detecting various types of malignant tumors, particular in patients with squamous cell carcinoma. PMID- 12548612 TI - Detection of chromosomal aneusomy by fluorescence in situ hybridization for patients with nipple discharge. AB - BACKGROUND: Breast carcinoma and precancer are believed to start in the lining of the milk duct or lobule. Ductography and fiberoptic ductoscopy (FDS) are used to identify abnormal intraductal lesions, although it is difficult to distinguish malignant from benign cases. Therefore, we studied the clinical usefulness of fluorescence in situ hybridization (FISH) analysis of a numerical aberration of chromosomes (aneusomy) using ductal lavage from patients with nipple discharge. METHODS: We applied ductography and FDS to 90 women who had nipple discharge. Ductal lavages obtained from patients with positive ductography and/or FDS findings were subjected to cytology and FISH analysis using centromere probes for chromosomes 1, 11, and 17. Patients with samples that showed aneusomy in at least one of the three chromosomes were diagnosed as positive. RESULTS: Histologic evaluation revealed 54 benign lesions and six malignancies. The sensitivity, specificity, and diagnostic accuracy were 33.3%, 88.9%, and 83.3%, respectively, for cytology and 100%, 100%, and 100%, respectively, for FISH. CONCLUSION: The results demonstrated that FISH has a diagnostic accuracy comparable to cytology. This technique has 100% specificity is making a definitive diagnosis of malignancy in patients with indeterminate cytologic results, suggesting that FISH diagnosis can be a good adjunct to cytology. PMID- 12548613 TI - Aberrant DNA methylation in pediatric patients with acute lymphocytic leukemia. AB - BACKGROUND: Aberrant methylation of promoter-associated cystosine-guanine (CpG) islands is an epigenetic modification of DNA frequently observed in adult patients with acute lymphocytic leukemia (ALL). This epigenetic modification has been associated with gene silencing, malignant transformation, and aging. It is not known whether there are epigenetic differences between pediatric patients and adult patients with ALL. METHODS: To investigate the methylation characteristics of pediatric patients with ALL and to determine whether DNA methylation can explain prognostic or biologic differences between pediatric and adult patients, the authors analyzed the methylation status of 7 promoter-associated CpG islands in 16 pediatric patients with ALL and compared them with the methylation characteristics of a cohort of adult patients with ALL. The genes analyzed included the estrogen receptor gene (ER), multidrug resistance gene 1 (MDR1), p15, C-ABL, CD10, p16, and p73. RESULTS: The mean methylation densities of ER, MDR1, CD10, p15, and C-ABL were 25.4%, 16.4%, 5.23%, 4.24%, and 4%, respectively. P16 was methylated in 11.7% of patients, and p73 was methylated in 17.6% of patients. One patient (6.2%) had methylation of 0 genes, 15 patients (93.7%) had methylation of >/= 1 gene, and 4 patients (25%) had methylation of 3-4 genes. Methylation of all these genes was < 2% (or methylation specific polymerase chain reaction negative) in nonneoplastic tissues. A significant inverse correlation was observed between methylation of CD10 and CD10 expression. No differences were observed between the methylation characteristics of pediatric patients and adult patients. CONCLUSIONS: The results indicate that DNA methylation is common in pediatric patients with ALL and that methylation of the genes studied does not account for prognostic differences between pediatric patients and adult patients with ALL. PMID- 12548615 TI - Breast carcinoma in women age 25 years or less. PMID- 12548614 TI - Protease-activated receptor (PAR)-1 and PAR-2 participate in the cell growth of alveolar capillary endothelium in primary lung adenocarcinomas. AB - BACKGROUND: Cell growth can be induced via elicitation of protease-activated receptors (PAR) with serine proteases such as thrombin and trypsin. METHODS: To understand whether PAR are involved in tumor vessel formation in the neoplastic cell-bearing alveolar walls, immunohistochemical and reverse transcriptase polymerase chain reaction analyses were performed using the lung tissues from 16 patients with primary lung adenocarcinomas. RESULTS: In microdissected tumor alveolar walls, the expressions of PAR-1 and PAR-2 mRNA were increased by 10-fold (P < 0.05) and 16-fold (P < 0.01), respectively, as compared with normal alveolar walls. Confocal microscopy revealed that tumor capillary endothelial cells in alveolar walls lost thrombomodulin expression. Instead, the expression of PAR-2 often became obvious at the normal border. Both PAR-1 and PAR-2 were expressed in the microvessel endothelial cells in tumors. Trypsin mRNA was expressed in 7 of the 16 cancer cell-bearing tissue specimens in contrast to 1 of the 14 normal alveolar walls. Immunohistochemically, trypsin was positive in the neoplastic cells from 10 patients and in lung adenocarcinoma cell lines (A549, HLC-1, LC-2, and PC-14). An in vitro assay showed a significant increase in idoxuridine (IdU) or bromodeoxyuridine uptake in human pulmonary artery endothelial cells and human umbilical cord vein endothelial cells after treatments with alpha-thrombin or activating peptides; SFLLRN for PAR-1 and SLIGKV for PAR-2, respectively. CONCLUSIONS: Thus, proliferation of alveolar capillary endothelial cells is initialized in part by PAR activation with serum thrombin and neoplastic cell released trypsin. These results suggest a synergistic effect of PAR with vascular endothelial growth factor in alveolar angiogenesis. PMID- 12548617 TI - Multicentric mammary carcinoma: evidence of monoclonal proliferation. PMID- 12548619 TI - Stepwise induced fit in the pico- to nanosecond time scale governs the complexation of the even-skipped transcriptional repressor homeodomain to DNA. AB - Induced fit effects in the complex of a DNA decamer with two even-skipped transcriptional repressor homeodomain molecules were investigated by means of molecular dynamics simulations. Dynamics of these effects are found to be in the time scale from pico- to nanoseconds. First steps are made by the fast-moving DNA backbone phosphates, which upon binding change their B(I)/B(II) substate distribution. Further rearrangements in the DNA double helix induced upon complexation, like bending of the helix axis, changes of the minor groove width, and of different helical parameters, are slower and occur within a few nanoseconds. The flexibility of the DNA, especially of its backbone, seems thereby to play an important role for specific DNA ligand recognition. PMID- 12548620 TI - Molecular mass and chain conformation of carboxymethylated derivatives of beta glucan from sclerotia of Pleurotus tuber-regium. AB - Seven water-insoluble (1 --> 3)-beta-D-glucan fractions TM8-1 to TM8-7 with weight-average molecular mass M(w) ranged from 2.22 to 77.4 x 10(4) obtained from the sclerotia of Pleurotus tuber-regium were carboxymethylated to produce the water-soluble fractions CTM8-1 to CTM8-7 with M(w) ranged from 3.87 to 87.8 x 10(4). The degree of substitution (DS) of CTM8 fractions was analyzed by ir and elemental analysis (EA) to be 0.3-0.68. The M(w) and the intrinsic viscosity [eta] of the CTM8 fractions were measured by size-exclusion chromatography combined with multiangle laser light scattering (SEC-MALLS), MALLS, and viscometry in phosphate buffer solution (PBS) at 37 degrees C. The dependencies of [eta] and radius of gyration (z) (1/2) on M(w) for the CTM8 samples were found to be [eta] = (8.82 +/- 0.03) x 10(-3) M(w)(0.78 +/- 0.04) (cm(3) g(-1)) and (z) (1/2) = (3.09 +/- 0.05) x 10(-3) M(w)(0.75 +/- 0.06) (nm) in the M(w) range from 3.87 x 10(4) to 53.2 x 10(4). Based on current theories for wormlike chain model, the conformational parameters of the CTM8 were obtained to be 790 (nm(-1)) for M(L), 9.6 (nm) for q, which were higher than those of the native TM8 fractions, suggesting a more extended flexible chain of CTM8 in PBS. On the whole, the CTM8 fractions showed higher antitumor activity than their corresponding TM8 fractions. In view of data from molecular parameters and bioactivity, the antitumor activity of the CTM8 fractions may be correlated to its water solubility and relatively extended chain. PMID- 12548621 TI - Hybrid Monte Carlo with multidimensional replica exchanges: conformational equilibria of the hypervariable regions of a llama VHH antibody domain. AB - Since the structural repertoire of the hypervariable regions of human antibodies is known to be more restricted than what is implied by sequence variability, a common approach to structural prediction is to use a knowledge-based (KB) method, such as the canonical structure model (C. Chothia and A. M. Lesk, Journal of Molecular Biology, 1987, Vol. 196, pp. 901-917). However, this model is less successful when applied to camelid heavy chain antibodies. In this study, molecular simulations were used to examine the conformational equilibria of the hypervariable regions (H1, H2, and H3) of a llama heavy chain variable domain, for which KB predictions are poor. Simulations were carried out using both conventional molecular dynamics (MD) and hybrid Monte Carlo with multidimensional replica exchanges (HYMREX). The advantage of the latter method is its ability to selectively target parts of the Hamiltonian that can most readily improve sampling. A novel variant of HYMREX was implemented in which, besides the temperature, torsional interactions and the range of nonbonded interactions were varied. To compare the sampling abilities of MD and this HYMREX scheme, simulations were started from a misfolded conformational state. Overall, MD yielded final conformations more similar to the initial state, implying quasi ergodic sampling. In contrast, HYMREX achieved more ergodic sampling, and the majority of conformations that it sampled agreed well with the known crystal structure. The HYMREX simulation results were used to help identify the chief interactions governing the conformational equilibria and to reexamine the key assumptions underlying the KB predictions. The data show that the H1 region exhibited significant conformational freedom, in support of the hypothesis that main-chain structural variability in this region could play a greater role in antigen binding in camelid antibodies than it does in normal antibodies. Key H1 residues and associated inter-loop interactions are conjectured to account for the poor KB predictions. PMID- 12548622 TI - Folding of peptides characterized by c3Val, a highly constrained analogue of valine. AB - Using a combined chemical/chiral chromatographic approach we synthesized an N protected derivative of (R)-c(3)Val, a severely conformationally restricted C(alpha)-tetrasubstituted alpha-amino acid characterized by a C(beta,beta) dimethylated cyclopropane system. A set of terminally protected derivatives and model peptides (to the heptamer level), containing one or two (R)-c(3)Val residues in combination with either Aib or Gly residues, was prepared by solution methods. A detailed solution and crystal-state conformational investigation, based on Fourier transform infrared (FTIR) absorption, (1)H-NMR, and x-ray diffraction techniques, performed in comparison with a similar study on related derivatives and peptides rich in (alphaMe)Val, the prototype of C(alpha) tetrasubstituted alpha-amino acids of this subfamily, allowed us to conclude the following: (a) c(3)Val is a good beta-bend and helix former, although less efficient than (alphaMe)Val. (b) The relationship between alpha-carbon chirality and screw sense of the folded structure formed is the same as that of (alphaMe)Val, i.e., the (R)-enantiomer has a strong left-handed bias. (c) c(3)Val seems more prone than (alphaMe)Val to fold into a gamma-bend conformation. The conformational propensities of C(beta,beta)-disubstituted Ac(3)c residues are also discussed in comparison with those of the parent cyclopropane residue. PMID- 12548623 TI - Simulation of the bis-(penicillamine) enkephalin in ammonium chloride solution: a comparison with sodium chloride. AB - In order to quantify specific ion effects, a simulation study of bis(penicllamine) enkephalin, also known as DPDPE, has been performed in aqueous ammonium chloride solution and has been compared to a previous simulation of DPDPE in aqueous sodium chloride solution. Global thermodynamics have been calculated for a model system and the solution environment around DPDPE has been characterized. Associations of ions with DPDPE have been investigated. The observed differences between sodium chloride solution and ammonium chloride solution suggest that individual cations affect the solvation and peptide binding properties of a given anion. PMID- 12548624 TI - The thermodynamic contribution of the 5-methyl group of thymine in the two- and three-stranded complexes formed by poly(dU) and poly(dT) with poly(dA). AB - To assess the thermodynamic contribution of the 5-methyl group of thymine, we have studied the two-stranded helical complexes poly(dA).poly(dU) and poly(dA).poly(dT) and the three-stranded complexes--poly(dA).2poly(dU), poly(dA).poly(dT).poly(dU) and poly(dA).2poly(dT)--by differential scanning calorimetry, and uv optical melting experiments. The thermodynamic quantities associated with the 3 --> 2, 2 --> 1, and 3 --> 1 melting transitions are found to vary with salt concentration and temperature in a more complex manner than commonly believed. The transition temperatures, T(m), are generally not linear in the logarithm of concentration or activity of NaCl. The change in enthalpy and in entropy upon melting varies with salt concentration and temperature, and a change in heat capacity accompanies each transition. The poly(dA).2poly(dU) triple helix is markedly different from poly(dA).2poly(dT) in both its CD spectrum and thermodynamic behavior, while the poly(dA).poly(dT).poly(dU) triple helix resembles poly(dA).2poly(dT) in these properties. In comparing poly(dA).2poly(dT) with either the poly(dA).poly(dT).poly(dU) or the poly(dA).2poly(dU) triplexes, the substitution of thymine for uracil in the third strand results in an enhancement of stability against the 3 --> 2 dissociation of deltadeltaG degrees = -135 +/- 85 cal (mol A)(-1) at 37 degrees C. This represents a doubling of the absolute stability toward dissociation compared to the triplexes with poly(dU) as the third strand. The poly (dA).poly (dT) duplex is more stable than poly(dA).poly(dU) by deltadeltaG degrees = -350 +/- 60 cal (mol base pair)(-1) at 37 degrees C. Poly(dA).poly(dT) has 50% greater stability than poly(dA).poly(dU) as a result of the dT for dU substitution in the duplex. PMID- 12548625 TI - Electron microscopy of functional ribosome complexes. AB - Cryoelectron microscopy has made a number of significant contributions to our understanding of the translation process. The method of single-particle reconstruction is particularly well suited for the study of the dynamics of ribosome-ligand interactions. This review follows the events of the functional cycle and discusses the findings in the context provided by the recently published x-ray structures. PMID- 12548626 TI - Assembly of the 30S ribosomal subunit. AB - Ribosomes are large macromolecular complexes responsible for cellular protein synthesis. The smallest known cytoplasmic ribosome is found in prokaryotic cells; these ribosomes are about 2.5 MDa and contain more than 4000 nucleotides of RNA and greater than 50 proteins. These components are distributed into two asymmetric subunits. Recent advances in structural studies of ribosomes and ribosomal subunits have revealed intimate details of the interactions within fully assembled particles. In contrast, many details of how these massive ribonucleoprotein complexes assemble remain elusive. The goal of this review is to discuss some crucial aspects of 30S ribosomal subunit assembly. PMID- 12548627 TI - Operator recognition by the phage 434 cI repressor: MD simulations of free and bound 50-bp DNA reveal important differences between the OR1 and OR2 sites. AB - Using molecular dynamics simulations in explicit solvent, we investigated the behavior of a 50-bp DNA sequence containing the 434 bacteriophage operators OR1 and OR2 separated by an 8-bp spacer. Two simulations of 1 ns each were carried out, with DNA alone and with DNA complexed to dimers of the R1-69 DNA binding domain of the phage 434 cI repressor protein at the OR1 and OR2 sites. Strong correlations among average structural parameters are observed between our simulations and available experimental data for the bound OR1/OR2 subsites. In the free state, some differences appear between the three relevant fragments (OR1, the spacer, and OR2). Unbound OR1 exhibits a large, shallow major groove into which the base atoms protrude and is also bent toward the major groove. This structure is maintained because structural fluctuations are weak. Unbound OR2 resembles canonical B-DNA although the structural parameters show greater fluctuations, essentially due to a malleable step (the innermost CpA/TpG), absent in OR1. Complexation with the proteins slightly alters the base positions but strongly modifies the sugar and backbone motions. The most crucial repressor effects are changes in the flexibility of the OR1/OR2 sites. Structural fluctuations are enhanced for OR1, conferring a favorable energetic contribution to the OR1 binding, whereas they are reduced for OR2. Therefore, both structural and dynamic properties of DNA suggest OR1 is the most attractive site for the repressor, which may explain the different binding association constants observed for the OR1 and OR2 sites. Finally, we also investigated the impact of the protein on the DNA backbone dynamics and find that direct or indirect interactions facilitate the DNA structural variations required for achieving complementarity with the protein. PMID- 12548628 TI - Surface electrostatic effects in oligonucleotide microarrays: control and optimization of binding thermodynamics. AB - We present a theoretical thermodynamic framework for the design of more efficient oligonucleotide microarrays. A general thermodynamic relation is derived to describe the electrostatic surface effects on the binding of the assayed biomolecule to a surface-tethered molecular probe. The relation is applied to analyze how the nucleic acid target, the oligonuleotide probe, and their DNA duplex electrostatic interactions with the surface affect the hybridization on DNA arrays. Taking advantage of a closed form exact solution of the linear Poisson-Boltzmann equation for a charged ion-penetrable sphere in electrolyte solution interacting with a plane wall, we study the effects of the surface and solution conditions. Binding free energy is found as a function of the surface material, dielectric or metal, the surface charge density, linker molecule length, temperature, and added salt content. The charge or electric potential of the dielectric or metal surface, respectively, is shown to dominate the hybridization, especially at low added salt or short linker length. We predict that substantial enhancement of sensitivity, selectivity, and reliability of microarrays can be achieved by control of the surface conditions. As examples, we discuss how to overcome two limitations of current technologies: nonequal sensitivity of the probes with different GC and AT bases content, and poor match/mismatch discrimination. In addition, we suggest the design of microarray conditions where the tested nucleic acid is unfolded, thus making possible the screening of a larger sequence with single nucleotide resolution. These promising findings are discussed and further experimental tests suggested. PMID- 12548629 TI - Protein arrays: the current state-of-the-art. AB - The completion of projects for sequencing whole genomes such as those of human, Saccaromyces cerevisiae and Caenorhabditis elegans has led to a rapid increase in the availability of genetic information. The technology by which such information is acquired is having a major impact on the way we think about analysing the RNA and protein products of the gene transcription. Transcriptome analysis has, via microarray technology, managed to achieve a near genomic scale quantitative analysis of mRNA levels. Several other technologies such as quantitative reverse transcriptase polymerase chain reaction and representational differential analysis are also available for analysing gene products. Traditionally, protein analysis has been performed by assaying one particular protein at a time, with very little parallel analysis. As protein function is a direct consequence of the protein product of the gene and as mRNA levels do not always correlate well with protein, it is desirable to analyse the entire protein complement of a cell etc. on a similar scale to mRNA. Despite the clear interest in analysing the expression, structure and function of proteins at a genomic scale, they have proven less amenable to such generic, high-throughput approaches. This review highlights the current thinking in the area of proteome analysis and considers the potential for future technology development in the area of protein arrays. PMID- 12548630 TI - Expanding the organismal scope of proteomics: cross-species protein identification by mass spectrometry and its implications. AB - Due to the limited applicability of conventional protein identification methods to the proteomes of organisms with unsequenced genomes, researchers have developed approaches to identify proteins using mass spectrometry and sequence similarity database searches. Both the integration of mass spectrometry with bioinformatics and genomic sequencing drive the expanding organismal scope of proteomics. PMID- 12548631 TI - Application of immunoproteomics to analysis of post-translational processing of the antiphagocytic M protein of Streptococcus. AB - Post-translational modification of the antiphagocytic M1 protein of Streptococcus pyogenes can influence its binding properties for human immunoglobulin G subclasses and its invasive potential. Current methods of monitoring this modification event involve N-terminal sequencing and are cumbersome, slow and not amenable to routine analysis. In this study we demonstrate that surface enhanced laser desorption/ionization-time of flight mass spectrometry can be used to monitor modification of the M1 protein by the secreted bacterial cysteine protease, SpeB. This method, when combined with a specific antibody capture step provides a specific, rapid and sensitive assay for key virulence factors of the important human pathogen Streptococcus pyogenes. PMID- 12548632 TI - A novel experimental design for comparative two-dimensional gel analysis: two dimensional difference gel electrophoresis incorporating a pooled internal standard. AB - The comparison of two-dimensional (2-D) gel images from different samples is an established method used to study differences in protein expression. Conventional methods rely on comparing images from at least 2 different gels. Due to the high variation between gels, detection and quantification of protein differences can be problematic. Two-dimensional difference gel electrophoresis (Ettan trade mark DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. In the application of DIGE different samples are labelled with mass and charge matched spectrally resolvable fluorescent dyes and are then separated on the same 2-D gel. Using an Escherichia coli lysate "spiked" with varying amounts of four different known proteins, we have tested a novel experimental design that exploits the sample multiplexing capabilities of DIGE, by including a standard sample in each gel. The standard sample comprises equal amounts of each sample to be compared and was found to improve the accuracy of protein quantification between samples from different gels allowing accurate detection of small differences in protein levels between samples. PMID- 12548633 TI - Serological and proteomic evaluation of antibody responses in the identification of tumor antigens in renal cell carcinoma. AB - Renal cell carcinoma (RCC) is relatively resistant to conventional chemotherapy and radiotherapy. However, reports of spontaneous regression along with promising results in clinical trials suggest that immunotherapuetic strategies may be of clinical benefit. Few RCC related antigens have been identified to date, and the technical difficulty and time constraints of current antigen identification techniques preclude the screening of large numbers of patients. A comparatively rapid strategy has been used to identify components of tumors that elicit an antibody response in the patient - the serological and proteomic evaluation of antibody responses (SPEAR) approach. This combines two-dimensional polyarylamide gel electrophoresis of tumor and normal kidney samples with immunoblotting using autologous patient sera and protein identification by mass spectrometry. Using the SPEAR approach to screen RCC patients for naturally occurring antitumor antibody responses, a number of candidate immunogens have been identified in patients with high-grade disease and their relative expression levels in tumor tissue compared to normal tissue have been studied. These proteins include annexins I and IV, thymidine phosphorylase (TP), carbonic anhydrase I, Mn superoxide dismutase and major vault protein (MVP). Downstream analysis of the tissue expression of some of these proteins shows that MVP is up-regulated in 2/4 of RCC tumors but is also expressed in normal kidney whereas TP is up-regulated in 100% (11/11) of RCC cases examined with no or minimal expression in normal kidney, indicating a potential use as a therapeutic target. PMID- 12548634 TI - Antibody microarray profiling of human prostate cancer sera: antibody screening and identification of potential biomarkers. AB - We developed a practical strategy for serum protein profiling using antibody microarrays and applied the method to the identification of potential biomarkers in prostate cancer serum. Protein abundances from 33 prostate cancer and 20 control serum samples were compared to abundances from a common reference pool using a two-color fluorescence assay. Robotically spotted microarrays containing 184 unique antibodies were prepared on two different substrates: polyacrylamide based hydrogels on glass and poly-1-lysine coated glass with a photoreactive cross-linking layer. The hydrogel substrate yielded an average six-fold higher signal-to-noise ratio than the other substrate, and detection of protein binding was possible from a greater number of antibodies using the hydrogels. A statistical filter based on the correlation of data from "reverse-labeled" experiment sets accurately predicted the agreement between the microarray measurements and enzyme-linked immunosorbent assay measurements, showing that this parameter can serve to screen for antibodies that are functional on microarrays. Having defined a set of reliable microarray measurements, we identified five proteins (von Willebrand Factor, immunoglobulinM, Alpha1 antichymotrypsin, Villin and immunoglobulinG) that had significantly different levels between the prostate cancer samples and the controls. These developments enable the immediate use of high-density antibody and protein microarrays in biomarker discovery studies. PMID- 12548635 TI - Proteome analysis on an early transformed human bronchial epithelial cell line, BEP2D, after alpha-particle irradiation. AB - To probe the mechanism of carcinogenesis of lung cancer at the molecular level and to find potential protein markers involved in the early phase of tumorgenesis, differential proteome analysis on primary passage cell line R15H, and early transformed cell line R15H20 derived from (238)Pu alpha-particle irradiation of human papillomavirus (HPV) 18-immortalized human bronchial epithelial cell line (BEP2D), was carried out using two-dimensional electrophoresis (2-DE) and peptide mass fingerprinting (PMF) with matrix-assisted laser desorption/ionisation-time of flight mass spectrometry. Image analysis and Student's t-test (p < 0.05) showed that three protein spots were only expressed in R15H, intensities of 43 protein spots on the gels were altered between R15H and R15H20. Two of the three spots that were only expressed in R15H were identified as high mobility group protein 1. Two proteins decreased in abundance in R15H20 were identified as maspin precursor, a tumor suppressor and aminoacylase-1. Ornithine aminotransferase and peptidyl-prolyl cis-trans isomerase A that were increased in R15H20, were also identified. Relationships between these differentially expressed proteins and the carcinogenesis mechanism of lung cancer are discussed. The protein expression profile of the R15H cell line was also constructed during the study as a reference map for further comparative proteome analysis of the irradiation induced BEP2D cell line. Of the 90 spots analyzed with PMF in the 2-DE gel of R15H cell line, 50 proteins were identified by searching the nonredundant protein database SWISS-PROT/TrEMBL. PMID- 12548636 TI - Vitamin E prevents oxidation of antiapoptotic proteins in neuronal cells. AB - Oxidative damage to neuronal proteins appears to be central to the toxicity associated with a number of neuropathologies, including Alzheimer's disease. We have examined this by using oxidative stress to induce apoptosis in a mouse hippocampal neuronal cell line (HT-22). Oxidatively modified proteins were measured by high-resolution two-dimensional gel electrophoresis coupled with oxidation-specific immunostains. Under these conditions the oxidatively stressed cells undergo apoptosis, and specific proteins are oxidized. The three proteins that appeared to be most susceptible to oxidation were identified by mass spectrometry. Those oxidized proteins are heat shock protein 60 and vimentin, both believed to function as antiapoptotic proteins, and a third protein with sequence homology to hemoglobin alpha-chain. When the cells were pretreated with vitamin E, these proteins were not oxidized and the cells did not undergo apoptosis. PMID- 12548637 TI - Proteome analysis of the thermoreceptive pit membrane of the western diamondback rattlesnake Crotalus atrox. AB - Rattlesnakes detect their prey's temperature by means of a cavern-like structure, the pit organ. The sensory component of this organ lies within a thin membrane called the pit membrane. Proteome analysis conducted on this neurosensory tissue revealed only a relatively small number of proteins, thereby depicting its high degree of specialization. In addition to containing blood serum and structural proteins, the proteome of this membrane appears to be strikingly similar to that of isolated rattlesnake brain mitochondria. Indeed, our results show that over 80% of the detected tissue proteins are of mitochondrial origin. Fluorescence microscopy studies of these organelles indicate their dense arrangement and accumulation in structures which have been previously reported to be the terminal ends of free nerve fibers of the innervating trigeminal branches. Thus, original ultrastructural observations are paralleled by our findings at the molecular level. PMID- 12548638 TI - Proteomic analysis of human bronchoalveolar lavage fluid: expression profiling of surfactant-associated protein A isomers derived from human pulmonary alveolar proteinosis using immunoaffinity detection. AB - Human bronchoalveolar lavage fluid (BALF) proteins from pulmonary alveolar proteinosis (PAP) obtained by washing the epithelial lining of the lung with phosphate-buffered saline, were separated using high resolution two-dimensional gel electrophoresis (2-DE) under denaturing and reducing conditions. By Western blotting, the proteins were transferred from polyacrylamide gel onto a chemical resilient membrane. The surfactant-associated protein A (SP-A) isomers were then identified with enhanced chemiluminescence detection (ECL) using antibody-antigen reaction. Some of the gels were treated with silver staining after 2-DE. The molecular masses of SP-A isomers in BALF from PAP ranged from 20.5 to 26, 26 to 32, and 32 to 42 kDa, respectively; and isoelectric points (pI) were in pH range of 4.5-5.4 under denaturing and reducing conditions. In the mass range of 20.5-26 kDa and pI of 4.5-5.4, there were five isomers, and in mass range of 26-32 kDa and pI of 4.5 to 5.4, there were at least eight isomers on the ECL detection film. However, in the mass range of 32-42 kDa and pI of 4.5-5.4, there were three isomers separated one from another but there was also a cluster of overlapping spots on the ECL detection film. Thus, this communication describes a characteristic 2-DE pattern of SP-A isomers in BALF from PAP as follows. (1) The five isomers of mass 20.5-26 kDa and pI of 4.5-5.4; (2) the eight isomers of mass 26-32 kDa and pI of 4.5-5.4; and (3) the three isomers of mass 32-42 kDa and pI of 4.5-5.4. PMID- 12548639 TI - Proteins induced by Xanthomonas axonopodis pv. passiflorae with leaf extract of the host plant (Passiflorae edulis). AB - Two-dimensional gel electrophoresis was used to identify differentially displayed proteins during treatment of Xanthomonas axonopodis pv. passiflorae in media containing leaf extract of the compatible (passion fruit) and incompatible (tomato) hosts. The results showed that at different times of treatment (5, 25 and 45 h) the global expression of proteins was almost identical in cells grown in minimal medium (MM) and in medium containing leaf extract of the incompatible host (MMT). The protein patterns of cells grown in medium containing passiflorae (MMP) leaf extract and MM were also compared enabling the detection of 17 differential spots. Most of the proteins were induced at earlier times of incubation (5 h) and maintained until 45 h in MMP. By using another carrier ampholyte range, seven additional proteins were identified in MMP treated cells. Five proteins, including one constitutive, two induced and two up-regulated in MMP were microsequenced. All sequences were found in the genome of xanthomonads sharing high level of identity (88-100%). Fructose biphosphate aldolase was expressed in all media employed. A putative membrane-related protein and a hypothetical protein were novel proteins induced specifically by the passiflorae extract. An inorganic pyrophosphatase and a hypothetical protein that showed similarity to the yciF gene of Salmonella thyphimurium were up-regulated in MMP. PMID- 12548644 TI - A comparative assessment of psychological and psychosocial characteristics of cancer patients and their caregivers. AB - We recruited 50 cancer patients and their caregivers with the aim of extending our knowledge of emotional, personality and psychosocial variables, and comparing their reciprocal experience of the disease. The patients and caregivers were administered four of the questionnaires included in the Cognitive Behavioral Assessment 2.0, the Family Strain Questionnaire and the Satisfaction with Life Scale. The patients were characterised by significantly greater emotional disturbances than their caregivers, who were emotionally stable and had a relatively low level of perceived strain despite their high level of hostility and state anxiety. The intra-scale correlations highlighted the fact that the perception of distress depends on neuroticism and the presence of anxiety and depression (which are themselves known to be connected with neuroticism). The everyday life of the caregivers seemed to be characterised by restricted social relationships and interests; a relatively large proportion also declared that they had practical problems related to disease management, economics and some embarrassing and stigmatic aspects of the disease itself. PMID- 12548645 TI - The mental adjustment to cancer (MAC) scale: French replication and assessment of positive and negative adjustment dimensions. AB - The Mental Adjustment to Cancer (MAC) scale was validated on a heterogeneous French sample of 317 cancer patients. Internal consistency was satisfactory for the original subscales (alpha coefficients=0.62-0.80), except for the Fatalism subscale (alpha=0.40). The intercorrelations of the subscales and the correlations between the subscales and Anxiety and Depression criteria were congruent with the values reported in the literature. Multidimensional Scaling revealed three positive and three negative subsets of items revealing adjustment to cancer. Congeneric factor analysis of the subsets was performed with LISREL 8.3 and only three of them (after discarding certain items) were retained: Fighting Spirit (FS) Hopelessness/Helplessness (HH) and Anxious Preoccupation (AP). A confirmatory hierarchical factor analysis on the 21 items included showed that FS measured positive adjustment to cancer and HH and AP measured negative adjustment. A differential adjustment hypothesis was proposed in order to explain the stability and instability of the measures of the diverse constructs. The three revised subscales showed the same validity pattern as the corresponding original scales, but the magnitude of the correlations was considerably improved with respect to the original subscales. The practical and the theoretical importance of FS, HH and AP are emphasized. PMID- 12548646 TI - Breast cancer patients' perceptions of their husbands' support in a cross cultural context. AB - In light of the Western focus in the breast cancer literature on spouse support, this study attempted to investigate ethnic differences in their perceptions of the support provided by their spouses. Forty-six women who had had breast cancer 6 months to 3 years prior participated in this study. These women were selected from three ethnic groups (13 Euro-American (EA) women, 18 Chinese-American women and 15 Japanese-American women). The study attempted to assess the women's perceptions at two levels of inquiry. One was based on standardized testing and scales and the second was a qualitative semi-structured interview. It was hypothesized that few differences would exist in standardized testing, and that cultural differences would be found at the level of semi-structured interviewing. Results showed that in fact few differences existed among the three groups on standardized instruments. At the level of qualitative evaluation, however, it appeared that Asian-American women were expected to be self-sacrificing and nurturing of husband and family, while EA women were able to be dependent. Another major difference existed around the theme of harmony and intimacy, where Asian-American women had a goal of harmony over intimacy, while EA women had a goal of intimacy over harmony. A final major difference was in the area of communication, with both Asian groups appearing to value non-verbal communication over verbal, while EA women appeared to value verbal communication over non verbal. PMID- 12548647 TI - Understanding the breast cancer experience of Asian American women. AB - Breast cancer is the most common form of cancer in women and affects women across all ethnic groups. Although the psychosocial impact of breast cancer is being studied, there is little information on ethnic minorities. To better understand the breast cancer experience of Asian American women, we conducted key informant and focus group interviews. Six professionals participated in the key informant interviews. A total of 34 Asian American breast cancer survivors participated in focus group interviews, including Korean (n=10), Chinese (n=11), and a mixed Asian group (n=13). The common themes identified in this series of qualitative studies included: lack of knowledge about breast cancer; medical care issues such as cost and amount of time spent with physician; cultural factors related to beliefs about illness, gender role and family obligations (e.g. self-sacrifice) and language barriers; the importance of spirituality; and psychosocial concerns related to worry about children, burdening the family, body image and sexual health concerns. A primary source of support and coping for Asian American women with breast cancer was their spiritual beliefs. The results from this qualitative study have been used to prepare a survey instrument to examine these issues in a larger sample of Asian American women. PMID- 12548648 TI - The relationship between cancer locus of control and complementary and alternative medicine use by women diagnosed with breast cancer. AB - Research has shown that perceptions of control influence health-related behavior and outcomes. This study explored the influence of the control constructs in the context of the theory of cognitive adaptation to the use of complementary and alternative medicine (CAM) among 551 women diagnosed with breast cancer in Portland, Oregon. The majority of these women had high perceptions of cancer control and used one or more types of CAM therapy. Multinomial logistical regression indicated that higher perceptions of control over the course and cause of cancer significantly predicted CAM use. The model combining sociodemographic variables (age, education and type of health insurance) and control variables explained more variation (R(2)=0.23) in predicting CAM use than the model with only sociodemographic variables (R(2)=0.14), indicating that perceptions of control over the cause and the course of cancer had an independent significant influence. It was concluded that the assessment of control constructs from the theory of cognitive adaptation is useful for studying CAM use among women diagnosed with breast cancer and that understanding the meanings that lie behind the decisions to use these therapies are essential for health care providers who are promoting the health and well-being of their patients. PMID- 12548649 TI - Patients' satisfaction ratings and their desire for care improvement across oncology settings from France, Italy, Poland and Sweden. AB - There has been an increasing interest in patient satisfaction assessment across nations recently. This paper reports on a cross-cultural comparison of the comprehensive assessment of satisfaction with care (CASC) response scales. We investigated what proportion of patients wanted care improvement for the same level of satisfaction across samples from oncology settings in France, Italy, Poland and Sweden, and whether age, gender, education level and type of items affected the relationships found. The CASC addresses patient's satisfaction with the care received in oncology hospitals. Patients are invited to rate aspects of care and to mention for each of these aspects, whether they would want improvement.One hundred and forty, 395, 186 and 133 consecutive patients were approached in oncology settings from France, Italy, Poland and Sweden, respectively. Across country settings, an increasing percentage of patients wanted care improvement for decreasing levels of satisfaction. However, in France a higher percentage of patients wanted care improvement for high-satisfaction ratings whereas in Poland a lower percentage of patients wanted care improvement for low-satisfaction ratings. Age and education level had a similar effect across countries. Confronting levels of satisfaction with desire for care improvement appeared useful in comprehending the meaning of response choice labels for the CASC across oncology settings from different linguistic and cultural background. Linguistic or socio-cultural differences were suggested for explaining discrepancies between countries. PMID- 12548650 TI - The emotional response of families to children with leukemia at the lower socio economic level in central Mexico: a preliminary report. AB - Little research has been done in developing countries on the emotional impact experienced by families who have a child diagnosed with leukemia. This preliminary study looked at parents in Mexico who had to cope with their child's leukemia in the face of meager financial and social resources. The 51 children in the study were under 15 years and being treated for leukemia in hospitals affiliated with the Mexican Social Security Institute (IMSS) where their parents were interviewed using a questionnaire to ascertain their emotional responses to the illness. The data are analyzed and reported in five domains: perceived illness; psychological impact; coping strategies; family relationships; socio economic impact. A strengthening of family bonds was found the most common response (82.4%). The second most common responses were concern for the expenses incurred by the illness and the time dedicated to caring for the sick child (both 78.4%). It is especially important to assess families with meager social and financial resources as to their emotional responses to life-threatening illness because these limitations impose greater burdens and make coping more difficult. Psychosocial interventions are key to ensuring adequate treatment of the child in these circumstances. PMID- 12548651 TI - Bone marrow donation in childhood: one donor's perspective. AB - Allogeneic bone marrow transplantation is an established treatment modality for leukemia, hematologic disorders and inborn errors of metabolism. while the focus of attention is the transplant recipient, the bone marrow donor (often a sibling) is both physically and emotionally involved in the process. Few reports have been written from the donor's perspective, however. One of the authors of this report (G.P.) was a marrow donor as a child, and he reflects on this experience at the completion of his first year of training in medical school. PMID- 12548657 TI - Polychlorinated biphenyls affect gene expression in the rabbit preimplantation embryo. AB - Polychlorinated biphenyls (PCBs) have been shown to be embryotoxic. The mechanism(s) of action is not clearly understood. The toxic effects could be either direct or indirect. Furthermore, PCB congeners vary in their toxic potential. They can be classified in coplanar PCBs binding to the transcription factor aryl hydrocarbon receptor (AhR), which induce subsequent changes in gene expression, and noncoplanar PCBs exhibiting AhR-independent effects. In order to investigate possible mechanisms, 5 and 6 days old preimplantation rabbit embryos were exposed in vitro to low levels of coplanar (PCB 77, 126, and 169) or noncoplanar PCBs (PCB 28, 52, 101, 118, 138, 153, and 180). The PCB effects were studied by semiquantitative RT-PCR analysis of AhR target genes (cytochrome P450 (CYP) 1A1, 1A2, UDP-glucuronosyl transferase 1, glutathione S-transferase pi1 and aldehyde dehydrogenase) and dioxin-responsive genes (IL 1beta, PAI 2, Cox 2, TGFalpha, EGF, erbB 1-4, c-fos, c-jun, HSP 90, cyclophilin 40), and by differential display (DD) RT-PCR. CYP 1B1 mRNA and AhR protein were localized by in situ hybridization and immunohistochemistry, respectively. From the AhR target genes studied only CYP 1B1, and cyclooxygenase 2 showed an increase in mRNA levels after coplanar and noncoplanar PCB. Interleukin 1beta and plasminogen activator inhibitor 2 were downregulated. CYP 1B1 mRNA showed a stage specific inducibility at day 6, but not at day 5. By DD RT-PCR we identified six new genes previously not reported to be regulated by PCBs. The mRNAs encoding the subunits 1, 2, 4, and 5 of the NADH ubiquinone oxidoreductase and beta-globin showed a decrease, whereas trichohyalin mRNA was increased after PCB exposure. Coplanar and noncoplanar PCB congeners elicited similar responses on the mRNA levels of the studied genes. Exposure to coplanar PCBs did not result in the AhR being translocated to the nucleus. Our results show that (i). PCBs induce changes in gene expression in rabbit day 5 and 6 preimplantation embryos and imply (ii). that the transcriptional changes observed were not mediated by the nuclear AhR. PMID- 12548658 TI - Expression of retinol-binding protein and cellular retinol-binding protein in the bovine ovary. AB - Retinol (vitamin A) is essential for reproduction, and retinoids have been suggested to play a role in ovarian steroidogenesis, oocyte maturation, and early embryonic development. Retinol is transported systemically and intercellularly by retinol-binding protein (RBP). Within the cell, cellular retinol-binding protein (CRBP) functions in retinol accumulation and metabolism. Since the actions of retinoids are mediated, in part, by retinoid-binding proteins, the objective of this study was to investigate cell-specific expression of RBP and CRBP in the bovine ovary. Immunocytochemical analysis (ICC) localized RBP to the thecal and granulosa cell layers of antral and preantral follicles with the most intense staining in the cells of large, healthy follicles. The tunica adventitia of arterial blood vessels also exhibited RBP staining. Immunostaining of CRBP was most intense in the granulosa cells of preantral follicles and present, but diminished, in thecal and granulosa cells of antral follicles. Within the corpus luteum, both proteins were observed in large luteal cells, but only RBP was observed in small luteal cells. Northern blot analysis demonstrated that thecal and granulosa cells from antral follicles and luteal tissue expressed RBP and CRBP mRNA. Synthesis and secretion of RBP by thecal cells, granulosa cells, and luteal cells were demonstrated by immune-complex precipitation of radiolabeled RBP from the medium of cultured cells or explants, followed by SDS-PAGE and fluorography. Follicular fluid was collected from small (<5 mm) and large (8-14 mm) follicles, pooled according to follicular size, and analyzed for retinol, RBP, estradiol-17beta, and progesterone. Concentrations of retinol, RBP, and estradiol were greater in the fluid of large follicles. Results demonstrate retinoid-binding protein expression by bovine ovaries and provide physical evidence that supports the concept that retinoids play a role in ovarian function. PMID- 12548659 TI - Acquisition of transcriptional competence in the 1-cell mouse embryo: requirement for recruitment of maternal mRNAs. AB - We previously suggested that the transcriptional machinery is rate-limiting for genome activation in the preimplantation mouse embryo (Aoki et al., 1997: Dev Biol 181:296-307), suggesting that genome activation requires the synthesis of some proteins following fertilization. To test this hypothesis, transcriptional activity was measured in 1-cell embryos in which protein synthesis was inhibited by cycloheximide from the time shortly after insemination. Transcription in both the male and female pronuclei, as assessed by BrUTP incorporation, was markedly inhibited when assayed 14 hr post-insemination. This result suggested that newly synthesized proteins derived from maternally recruited mRNA, and not maternally derived proteins that were post-translationally modified following fertilization/egg activation, were critical for the acquisition of transcriptional competence. To test this hypothesis, mobilization of maternally recruited mRNAs was inhibited by treating the embryos with 3'-deoxyadenosine (3' dA), which prevents polyadenylation of mRNA. The results demonstrated that the transcriptional activity was markedly decreased in the embryos cultured with 2 mM 3'-dA, whereas 3'-deoxyguanosine had little inhibitory effect, and suggest that recruitment of maternal mRNAs is essential for acquisition of transcriptional competence. PMID- 12548660 TI - Expression of growth hormone and its transcription factor, Pit-1, in early bovine development. AB - During bovine embryogenesis, bovine growth hormone (bGH) contributes to proliferation, differentiation, and modulation of embryo metabolism. Pituitary specific transcription factor-1 (Pit-1) is a transcription factor that binds to promoters of GH, prolactin (PRL), and thyroid-stimulating hormone-beta (TSHbeta) encoding genes. A polymorphism in the fifth exon of the bGH gene resulting in a leucine (Leu) to valine (Val) substitution provides an Alu I restriction site when the Leu allele is present. To determine the onset of embryonic expression of the bGH gene, oocytes derived from ovaries homozygous for Leu alleles were fertilized in vitro with spermatozoa obtained from a Val homozygote. For each developmental stage examined, three separate pools of embryos composed of approximately 100 cell samples underwent RNA isolation, reverse transcription to cDNA, and amplification by nested PCR (nPCR). Bovine GH gene transcripts were identified at 2- to 4-cell (n = 162), 8- to 16-cell (n = 73), morulae (n = 51), and blastocyst (n = 15) stages. Likewise, transcripts for Pit-1 were detected at 2-cell (n = 125), 4-cell (n = 114), 8-cell (n = 56), 12-to-32-cell (n = 32), morulae (n = 68), and blastocyst (n = 14) stages. After digestion with Alu1, bGH cDNA was genotyped by restriction fragment length polymorphism (RFLP) analysis. Bovine GH mRNA was present in all pools of stages examined. Both Leu and Val alleles (maternal and paternal) were only detected in pools of embryos that had reached 8- to 16-cell stage. Results suggest that transcription of the bGH gene begins at the 8- to 16-cell stage in bovine embryos, possibly under control of the transcription factor, Pit-1, and that RFLP analysis of the bGH gene can be used to determine parental origin of transcripts in early embryonic development. PMID- 12548661 TI - Sperm mediated gene transfer in pig: Selection of donor boars and optimization of DNA uptake. AB - Transgenic animals are produced primarily by microinjecting exogenous DNA into the male pronuclei of a zygote. Microinjection is successful in mice but not efficient in farm animals, limiting its general utility. We have pursued an alternative technology for producing transgenic animals: Sperm Mediated Gene Transfer (SMGT). Based on our finding that sperm cells bind and internalize exogenous DNA, we used sperm as a vector for transmitting, not only their own DNA, but also, the exogenously-introduced gene of interest to the zygote. SMGT is highly efficient (up to greater than 80%) and relatively inexpensive; it can be used in species refractory to microinjection, whenever reproduction is mediated by gametes. In this report, we describe the procedure for selection of sperm donors and optimization of DNA uptake that are the key steps for the successful outcome of SMGT. We found that the nominal parameters that boar sperm should possess to serve as a good vector for exogenous DNA are the quality of semen based on standard parameters used in conventional animal breeding programs (volume, concentration, presence of abnormal sperm cells, motility at time of collection, and high progressive motility after 2 hr) and the ability of the sperm cells to take up and internalize exogenous DNA. The results described provide significant advances in SMGT technology applied to pigs, so that transgenic pigs can be efficiently obtained. Mol. PMID- 12548662 TI - Cleavage stage porcine embryos may have differing developmental requirements for karyopherins alpha2 and alpha3. AB - Numerous cellular proteins are able to localize to the nucleus due to the fact that they possess a nuclear localization signal (NLS) in their amino acid sequence. Nuclear localization sequences recognized by the importin alpha/beta heterodimer are found in cellular proteins capable of performing many diverse functions, ranging from chromatin remodeling to cell cycle regulation. Evidence has been presented that suggests individual importin alpha homologues are present at varying levels in different adult tissues. Other data have shown that specific subsets of NLSs found in different cellular proteins are recognized by individual importin alpha homologues with varying affinities. This evidence led us to hypothesize that due to the specific cargoes they carry, the mammalian embryo has different developmental requirements for individual importin alpha homologues. The results of the studies presented here indicate that importin alpha/beta mediated import occurs throughout early cleavage in the porcine embryo, as determined by a reporter protein microinjection assay, and that multiple importin alpha homologues are present throughout early cleavage, as determined by immunocytochemical analysis. An RNA interference approach was used in an attempt to determine the developmental requirements for specific importin alpha homologues during early cleavage in the porcine embryo. Results from this study showed that fertilized porcine embryos injected with double stranded RNA (dsRNA) corresponding to the importin alpha homologue karyopherin alpha3 had significantly fewer nuclei following four days of culture than did embryos injected with dsRNA for another importin alpha homologue, karyopherin alpha2, or two control groups. This is the first report indicating that mammalian embryos may have differential developmental requirements for specific nuclear trafficking pathways. PMID- 12548663 TI - Increase in macrophages in the testis of cathepsin a deficient mice suggests an important role for these cells in the interstitial space of this tissue. AB - Cathepsin A (PPCA) is a lysosomal carboxypeptidase that functions as a protective protein for alpha-neuraminidase and beta-galactosidase in a multienzyme complex. In the present study, the testes of PPCA -/- mice from 2 to 10 months of age were compared with those of their wild type counterparts. While germ and Sertoli cells appeared comparable in appearance and distribution, the mean profile area of seminiferous tubules showed a significant decrease between wild type and PPCA -/- mice, suggesting changes to the seminiferous tubules and their contents. In addition, macrophages in the interstitial space (IS) of PPCA -/- mice were large, spherical, and filled with pale lysosomes, unlike those seen in wild type mice, and a quantitative analysis of their frequency per unit area of IS in PPCA -/- mice revealed a significant increase compared to that of wild type mice; this was also the case for their mean profile area. Absence of mitotic figures, cycling cells, or degenerating figures in the IS suggests that the major recruitment of macrophages appears to be from the circulation. In the IS, Leydig cells also showed an accumulation of large pale lysosomes in PPCA -/- mice, and their frequency also increased significantly as compared to wild type mice. In the electron microscope, a close association of Leydig cell microvilli with the surface of macrophages was pronounced in PPCA -/- mice. Since macrophages and Leydig cells interact by secreting various factors between each other, and considering the fact that Leydig cells show an accumulation of large pale lysosomes in PPCA -/- mice, it is suggested that macrophages accumulate as a result of abnormalities occurring in Leydig cells. Taken together, the data on increase in frequency of macrophages suggests important functions for these cells in both wild type and PPCA -/- mice. PMID- 12548664 TI - Spherical bodies present within the germinal vesicle of Podarcis sicula previtellogenic oocyte derive from the temporaneous inactivation of ribosomal genes. AB - In the present paper we have investigated the origin of the spherical bodies (SBs) present within the germinal vesicle of about 400 microm previtellogenic oocytes in the lizard Podarcis sicula. In particular, we have attempted to clarify whether they derive from the single, large nucleolus present in early diplotenic oocyte as a consequence of ribosomal gene inactivation. We have, therefore, experimentally induced a decrease in rRNA synthesis by injecting animals with D-galactosamine or by exposing them to low temperatures. The investigations carried out have demonstrated that both treatments induce significant ultrastructural changes in the nucleolar apparatus and in particular fragmentation and the formation of SBs comparable to those observed in germinal vesicle under physiological conditions. These results indicate that the germinal vesicle of Podarcis sicula has a nucleolar apparatus that significantly changes its aspect according to its functional status and reveal that in this species, the time course of rRNA synthesis is peculiar with respect to any other vertebrate oocyte studies so far. PMID- 12548666 TI - Apoptosis during spermatogenesis in the spotted ray Torpedo marmorata. AB - This article is a cytological and molecular investigation on the occurrence of apoptosis during spermatogenesis in Torpedo, a cartilaginous fish characterised by a typical cystic testis. Using DNA fragmentation and Bak gene expression, it demonstrated that germ cells undergo apoptosis only at the stages of spermatocyte and spermatid, and degeneration also involves Sertoli but not Leydig cells. In immature cysts, this cellular process probably occurs when the ratio of germ cells to the only Sertoli cell (SC) forming the spermatoblast changes. Apoptosis also takes place in mature cysts after sperm release to eliminate most of the SCs. Few of them, however, become cytoplasts and probably continue secreting androgens so as to control the final events of spermatogenesis, i.e., passage of spermatozoa through the ductus deferentes. Finally, the present investigation demonstrated that, in Torpedo testis, Bak mRNA is expressed during spermatogenesis, thus suggesting that the mitochondrial pathway might be active. This observation in one of the oldest vertebrate classes indicates that, in all vertebrates, the apoptotic process during spermatogenesis is conserved, contributing to testicular homeostasis. PMID- 12548665 TI - Instability of crab vitellogenin and its immunological relatedness with mammalian atherogenic lipoproteins. AB - Vitellogenesis is the process of accumulation of vitellogenin (Vg) in rapidly growing oocytes of oviparous animals and its' subsequent transformation into lipovitellin (Lv). Lipovitellin, which forms the major yolk protein, serves as a principal nutrient reserve for the developing embryo. In the present study, Vg and Lv were purified from the hemolymph and ovary, respectively of the crab Scylla serrata by gel filtration followed by preparative gel electrophoresis. It was observed that purified Vg, but not Lv, possessed an intrinsic protease activity with which it underwent autoproteolysis giving rise to several smaller proteins. Furthermore, urea-mediated unfolding studies by UV-spectral analysis revealed clearly that Vg was easily disrupted by urea whereas Lv was resistant. Taken together, these results suggest that although Lv had a stable conformation, its precursor Vg was labile and highly sensitive to degradation. Another aspect that was investigated in the present study was the immunological kinship of crab Vg and Lv to mammalian atherogenic lipoproteins, the low density lipoprotein (LDL), very low density lipoprotein (VLDL), and apolipoprotein B (apoB). By Western blot analysis, it was demonstrated that crab Vg and Lv were immunoreactive to antibodies to human LDL, VLDL, and apoB. These observations suggest the existence of common epitope recognition sites in crab Vg and mammalian lipid transferring proteins. This corroborates well with our earlier study on the recognition of crab Vg receptor by mammalian lipoproteins. PMID- 12548667 TI - Glucose- and fructose-induced dog-sperm glycogen synthesis shows specific changes in the location of the sperm glycogen deposition. AB - Immunocytochemistry of glycogen deposition in dog sperm from fresh ejaculates showed that 53.7% of the population had glycogen at the midpiece, whereas 27.3% of the whole population accumulated glycogen only in the post-acrosomal region of the head. A similar distribution was observed when glycogen synthase, the enzyme which controls glycogen deposition, was studied. Incubation of spermatozoa with fructose increases the intracellular glycogen levels in a time- and concentration dependent manner. Moreover, after incubation with 2 mM fructose for 30 min, 47.3% of the spermatozoa had glycogen located in both the midpiece and post-acrosomal zone, 25.4% of the population showed the polysaccharide only in the midpiece, and 24.7% of the spermatozoa showed a uniform distribution of glycogen all over the cell. Similar results were observed after incubation with 10-mM fructose. The distribution of glycogen synthase followed a similar pattern to that of glycogen. Incubation with glucose also induced a time- and concentration-dependent increase of glycogen content, whereas incubation with 2 mM glucose for 30 min showed that the majority of the population (81.2%) had the glycogen distributed throughout either the midpiece or the midpiece and the post-acrosomal zones together. There were practically no cells with a uniform glycogen distribution. Similar results were obtained after incubation with 10-mM glucose, whereas glycogen synthase suffered a similar glucose-induced distribution change. These results indicate that dog-sperm glycogen metabolism is modulated by changes in the activity of their controlling enzymes and also by changes in the specific location places where glycogen synthesis is produced. PMID- 12548668 TI - Developmental failure of hybrid embryos generated by in vitro fertilization of water buffalo (Bubalus bubalis) oocyte with bovine spermatozoa. AB - The developmental potential of inter-species hybrid embryos produced by in vitro fertilization of in vitro matured buffalo oocytes with bovine spermatozoa was studied with a view to investigate pre-implantation embryo development and its gross morphology, early embryonic gene expression, and embryonic genome activation. Fertilization events with both buffalo and cattle spermatozoa were almost similar. Overall fertilization rate with cattle spermatozoa was 78.4% was not significantly different from that of buffalo spermatozoa (80.2%). Initial cleavage rate between buffalo and hybrid embryo was also similar, and there was no significant difference in their developmental rate till 8-cell stage (26.0 +/- 4.1 vs. 24.3 +/- 4.8). However, only 5.3% of hybrid embryos developed into blastocyst stage compared to 21.7% in buffalo. mRNA phenotyping of insulin-like growth factor family (Insulin, insulin receptor, IGF-I, IGF-I receptor, IGF-II, and IGF-II receptor) and glucose transporter isoforms (GLUT-I, II, III, IV) in hybrid embryos clearly showed that these molecules were not expressed after 8 cell stage onward. Similarly, as observed in buffalo embryos, incorporation of (35)S-methionine and (3)H-uridine could not be observed in hybrid embryos from 8 cell stage onward. This suggests that the maternal-zygotic genome activation did not occur in hybrid embryos. Differential staining also showed that the blastomere stopped dividing after 8-cell stage. Collectively, these parameters clearly showed that there was developmental failure of hybrid embryos. PMID- 12548669 TI - Ultrastructural modifications in bovine oocytes maintained in meiotic arrest in vitro using roscovitine or butyrolactone. AB - Butyrolactone-I (BL-I) and roscovitine (ROSC) are selective inhibitors of the cyclin-dependent kinases, and both have been shown to reversibly inhibit meiotic resumption in cattle oocytes for 24 hr without having a negative affect on subsequent development to the blastocyst stage. The aim of the present study was to describe the morphological changes occurring in fully grown immature and in vitro matured bovine oocytes following exposure to either BL-I or ROSC for 24 hr at concentrations known to be consistent with normal development. Immature bovine cumulus oocyte complexes, recovered from the ovaries of slaughtered heifers, were incubated for 24 hr in the presence of one of the inhibitors. They were then either fixed immediately and processed for transmission electron microscopy (TEM), or cultured for a further 24 hr in the absence of the inhibitor, in conditions permissive to maturation, and subsequently processed for TEM. A control group of oocytes were processed for TEM immediately upon recovery (0 hr) or following in vitro maturation (IVM) for 24 hr. In general, incubation with either inhibitor disrupted the integrity of the surrounding cumulus cells and affected their subsequent expansion during IVM. Within the oocyte cytoplasm, swelling of the mitochondrial cristae was immediately noticeable following meiotic inhibition in the presence of ROSC, while an increased population of pleomorphic mitochondria and mitochondria with electron lucent matrices following BL-I treatment was not observed until after the subsequent IVM period. Both inhibitors caused degeneration of the cortical granules, effectively reducing the population, most noticeably following IVM. At the level of the nucleus, both inhibitory treatments caused convolution of the nuclear membrane, furthermore, aberrant structures were observed within the nucleoplasm of ROSC-treated cumulus oocyte complexes (COCs). In conclusion, while it has been shown that inhibition of meiotic resumption using specific cdk inhibitors is possible and that such oocytes are capable of undergoing maturation, fertilization, and early embryo development, there is as yet no definitive proof that oocytes treated in this way can ultimately give rise to normal offspring. We have shown here that some modifications are induced in the oocytes at the ultrastructural level. Whether or not these modifications are compatible with normal gestation and the birth of a live calf remain to be elucidated. PMID- 12548670 TI - Review of proteomics with applications to genetic epidemiology. AB - Mapping of the human genome has the potential to transform the traditional methods of genetic epidemiology. The complete draft sequence of the 3.3 billion nucleotides comprising the genome is now available over the Internet, including the location and nearly complete sequence of the 26,000 to 31,000 protein encoding genes. However, aside from water, almost everything in the human body is either made of, or by, proteins. Although the DNA code provides the instructions for their amino acid sequence, there are an estimated 1.5 million proteins. Thus, the correlation between DNA sequence and protein is low, reflecting alternate splicing as well as post-translational modification. The purpose of this article is to explore ways in which the emerging field of proteomics, the study of proteins in a cell, may inform our approach to gene mapping. This article reviews the various technical approaches currently available for proteomics. Technologies are available to quantify protein expression (and compare normal versus disease states), identify proteins through comparison with sequence information in databases or direct sequencing (which can then be mapped to chromosomal locations to ensure appropriate markers), elucidate protein-protein interactions (which may underlie disease), determine localization of proteins within the cell (abnormal trafficking of proteins could have an inherited basis), and characterize modifications of proteins (which is relevant to modifier gene candidates). Several examples are presented to illustrate the potential application of proteomics to the field of genetic epidemiology, and we conclude with various considerations regarding design and analysis. PMID- 12548671 TI - Limits of fine-mapping a quantitative trait. AB - Once a significant linkage is found, an important goal is reducing the error in the estimated location of the linked locus. A common approach to reducing location error, called fine-mapping, is the genotyping of additional markers in the linked region to increase the genetic information. The utility of fine mapping for quantitative trait linkage analysis is largely unknown. To explore this issue, we performed a fine-mapping simulation in which the region containing a significant linkage at a 10-centiMorgan (cM) resolution was fine-mapped at 2, 1, and 0.5 cM. We simulated six quantitative trait models in which the proportion of variation due to the quantitative trait locus (QTL) ranged from 0.20-0.90. We used four sampling designs that were all combinations of 100 and 200 families of sizes 5 and 7. Variance components linkage analysis (Genehunter) was performed until 1,000 replicates were found with a maximum lodscore greater than 3.0. For each of these 1,000 replications, we repeated the linkage analysis three times: once for each of the fine-map resolutions. For the most realistic model, reduction in the average location error ranged from 3-15% for 2-cM fine-mapping and from 3-18% for 1-cM fine-mapping, depending on the number of families and family size. Fine-mapping at 0.5 cM did not differ from the 1-cM results. Thus, if the QTL accounts for a small proportion of the variation, as is the case for realistic traits, fine-mapping has little value. PMID- 12548672 TI - Multipoint affected sibpair linkage methods for localizing susceptibility genes of complex diseases. AB - Recently, Liang et al. ([2001] Hum. Hered. 51:64-78) proposed a general multipoint linkage method for estimating the chromosomal position of a putative susceptibility locus. Their technique is computationally simple and does not require specification of penetrance or a mode of inheritance. In complex genetic diseases, covariate data may be available which reflect etiologic or locus heterogeneity. We developed approaches to incorporating covariates into the method of Liang et al. ([2001] Hum. Hered. 51:64-78) with particular attention to exploiting age-at-onset information. The results of simulation studies, and a worked data example using a family data set ascertained through probands with schizophrenia, suggest that utilizing covariate information can yield substantial efficiency gains in localizing susceptibility genes. PMID- 12548673 TI - Genetic association tests with age at onset. AB - Many diseases or traits exhibit a varying age at onset. Recent data examples of prostate cancer and childhood diabetes show that compared to simply treating the disease outcome as affected vs. unaffected, incorporation of age-at-onset information into the transmission/disequilibrium type of test (TDT) does not appear to change the results much. In this paper, we evaluate the power of TDT as a function of age at onset, and show that age-at-onset information is most useful when the disease is common, or the relative risk associated with the high-risk genotype varies with age. Moreover, an extremely old unaffected subject can contribute substantially to the power of the TDT, sometimes as much as old-onset subjects. We propose a modified test statistic for testing no association between the marker at the candidate locus and age at onset. The simulation study was conducted to evaluate the finite sample properties of proposed and the TDT test statistics under various sampling schemes for trios of parents and offspring, as well as for sibling clusters where unaffected siblings were used as controls. PMID- 12548674 TI - Multivariate and multilocus variance components method, based on structural relationships to assess quantitative trait linkage via SEGPATH. AB - A general-purpose modeling framework for performing path and segregation analysis jointly, called SEGPATH (Province and Rao [1995] Stat. Med. 7:185-198), has been extended to cover "model-free" robust, variance-components linkage analysis, based on identity-by-descent (IBD) sharing. These extended models can be used to analyze linkage to a single marker or to perform multipoint linkage analysis, with a single phenotype or multivariate vector of phenotypes, in pedigrees. Within a single, consistent approach, SEGPATH models can perform segregation analysis, path analysis, linkage analysis, or combinations thereof. SEGPATH models can incorporate environmental or other measured covariate fixed effects (including measured genotypes), genotype-specific covariate effects, population heterogeneity models, repeated-measures models, longitudinal models, autoregressive models, developmental models, gene-by-environment interaction models, etc., with or without linkage components. The data analyzed can have any missing value structure (assumed missing at random), with entire individuals missing, or missing on one or more measurements. Corrections for ascertainment can be made on a vector of phenotypes and/or other measures. Because of the flexibility of the class of models, the SEGPATH approach can also be used in nongenetic applications where there is a hierarchical structure, such as longitudinal, repeated-measures, time series, or nested models. A variety of specific models are provided, as well as some comparisons with other linkage analysis models. Particular applications demonstrate the importance of correctly accounting for the extraneous sources of familial resemblance, as can be done easily with these SEGPATH models, so as to give added power to detect linkage as well as to protect against spuriously inferring linkage. PMID- 12548675 TI - Three-state frailty model for age at onset of dementia and death in Swedish twins. AB - We present a frailty model to estimate the relative importance of genetic and environmental factors on age at onset of dementia in a twin design. We use modern survival methodology to define a model that accounts simultaneously for longitudinal aspects, e.g., left truncation and right censoring in data, and the multivariate nature of twin data. Additionally, we present a novel three-state frailty model, with nondemented, demented, and dead states, describing variation in the onset of disease and mortality simultaneously in one model, while accounting for possible dependence for the two competing events. The frailty structure, i.e., the latent random effects structure, mimics the traditional twin model for continuous variables used in quantitative genetics, and as such describes within-pair dependence. This in turn leads to estimates for intrapair correlations, as well as for additive genetic, and shared and nonshared environmental components of variance. A hierarchical Bayesian model formulation and Gibbs sampling are used to estimate posterior distributions of the parameters. The models are applied to Swedish Twin Registry data on the onset of dementia in elderly twins. Based on the three-state frailty model, we estimate the intrapair correlations for dementia to be 0.87 [90% credible interval: 0.61,0.98] and 0.68[0.18,0.91] for MZ and DZ twins, respectively. Based on our model, we estimate that genetic effects account for about one third, and shared environmental effects for almost a half, of the variation in dementia hazards between individuals. More data, however, are needed to gain precision in these estimates. PMID- 12548680 TI - New insight into the transport mechanism of hydrated hydroxide ions in water. PMID- 12548676 TI - Power of multifactor dimensionality reduction for detecting gene-gene interactions in the presence of genotyping error, missing data, phenocopy, and genetic heterogeneity. AB - The identification and characterization of genes that influence the risk of common, complex multifactorial diseases, primarily through interactions with other genes and other environmental factors, remains a statistical and computational challenge in genetic epidemiology. This challenge is partly due to the limitations of parametric statistical methods for detecting genetic effects that are dependent solely or partially on interactions with other genes and environmental exposures. We previously introduced multifactor dimensionality reduction (MDR) as a method for reducing the dimensionality of multilocus genotype information to improve the identification of polymorphism combinations associated with disease risk. The MDR approach is nonparametric (i.e., no hypothesis about the value of a statistical parameter is made), is model-free (i.e., assumes no particular inheritance model), and is directly applicable to case-control and discordant sib-pair study designs. Both empirical and theoretical studies suggest that MDR has excellent power for identifying high order gene-gene interactions. However, the power of MDR for identifying gene-gene interactions in the presence of common sources of noise is not currently known. The goal of this study was to evaluate the power of MDR for identifying gene-gene interactions in the presence of noise due to genotyping error, missing data, phenocopy, and genetic or locus heterogeneity. Using simulated data, we show that MDR has high power to identify gene-gene interactions in the presence of 5% genotyping error, 5% missing data, or a combination of both. However, MDR has reduced power for some models in the presence of 50% phenocopy, and very limited power in the presence of 50% genetic heterogeneity. Extending MDR to address genetic heterogeneity should be a priority for the continued methodological development of this new approach. PMID- 12548681 TI - Diversified world: drive to life's origin?! PMID- 12548682 TI - Quantum tunneling of magnetization and related phenomena in molecular materials. AB - Molecules comprising a large number of coupled paramagnetic centers are attracting much interest because they may show properties which are intermediate between those of simple paramagnets and classical bulk magnets and provide unambiguous evidence of quantum size effects in magnets. To date, two cluster families, usually referred to as Mn12 and Fe8, have been used to test theories. However, it is reasonable to predict that other classes of molecules will be discovered which have similar or superior properties. To do this it is necessary that synthetic chemists have a good understanding of the correlation between the structure and properties of the molecules, for this it is necessary that concepts such as quantum tunneling, quantum coherence, quantum oscillations are understood. The goal of this article is to review the fundamental concepts needed to understand quantum size effects in molecular magnets and to critically report what has been done in the field to date. PMID- 12548683 TI - Ultrafast decarboxylation of organic peroxides in solution: interplay of different spectroscopic techniques, quantum chemistry, and theoretical modeling. PMID- 12548685 TI - Development of a new catalyst for the distannation of alkynes. PMID- 12548684 TI - Loss of isotope labeling in the conversion of [18O2]benzoic acid into [18O]benzoyl chloride with oxalyl chloride. PMID- 12548686 TI - A catalytic approach to (R)-(+)-muscopyridine with integrated "self-clearance". PMID- 12548687 TI - Gold dichloride and gold dibromide with gold atoms in three different oxidation states. PMID- 12548688 TI - Amplification of chirality from extremely low to greater than 99.5 % ee by asymmetric autocatalysis. PMID- 12548689 TI - Open network architectures from the self-assembly of AgNO3 and 5,10,15,20-tetra(4 pyridyl)porphyrin (H2tpyp) building blocks: the exceptional self-penetrating topology of the 3D network of [Ag8(ZnIItpyp)7(H2O)2](NO3)8. PMID- 12548690 TI - [Pd30(CO)26(PEt3)10] and [Pd54(CO)40(PEt3)14]: generation of nanosized Pd30- and Pd54-core geometries containing interpenetrating cuboctahedral-based metal polyhedra. PMID- 12548691 TI - A chemical adaptor system designed to link a tumor-targeting device with a prodrug and an enzymatic trigger. PMID- 12548692 TI - Gelation-assisted light harvesting by selective energy transfer from an oligo(p phenylenevinylene)-based self-assembly to an organic dye. PMID- 12548693 TI - Nucleotide cross-linking induced by photoreactions of platinum(IV)-azide complexes. PMID- 12548694 TI - Nitrido ruthenium porphyrins: synthesis, characterization, and amination reactions with hydrocarbon or silyl enol ethers. PMID- 12548695 TI - Stereocontrolled total synthesis of (+)-leucascandrolide A. PMID- 12548696 TI - Controlled formation of highly ordered cubic and hexagonal mesoporous nanocrystalline yttria-zirconia and ceria-zirconia thin films exhibiting high thermal stability. PMID- 12548697 TI - Thioglycoligases: mutant glycosidases for thioglycoside synthesis. PMID- 12548698 TI - Salinosporamide A: a highly cytotoxic proteasome inhibitor from a novel microbial source, a marine bacterium of the new genus salinospora. PMID- 12548699 TI - Multiple bonding between silicon and molybdenum: a transition-metal complex with considerable silylyne character. PMID- 12548701 TI - Issues concerning the construction of a metabolic model for neuronal activation. AB - The metabolic events underlying neuronal activity still remain the object of intense debate, in spite of the considerable amount of information provided from different experimental techniques. Indeed, several attempts at linking the cellular metabolic phenomena with the macroscopic physiological changes have not yet attained foolproof conclusions. The difficulties in drawing definitive conclusions are due primarily to the heterogeneity of the experimental procedures used in different laboratories, and also given the impossibility of extrapolating the findings obtained under stationary conditions (prolonged stimulation) to dynamic and transient phenomena. Recently, lactate has received much attention, following its proposal by Pellerin and Magistretti (1994; Proc. Natl. Acad. Sci. USA 91:10625-10629), instead of glucose, as the main substrate for neurons during activity. Several challenging aspects suggest the return to a more conventional view of neuronal metabolism, in which neurons are able to metabolize ambient glucose directly as their major substrate, also during activation. PMID- 12548702 TI - Neuronal distribution of EHSH1/intersectin: molecular linker between clathrin mediated endocytosis and signaling pathways. AB - Recent research indicates that the novel multimodular adaptor protein EHSH1 plays an important role in a partnership between clathrin-mediated endocytosis and intracellular signaling pathways, including the MAPK pathway, receptor-tyrosine kinase/ras-mediated pathway, and the rho family of the GTPase-dependent pathway. We report the detailed expression pattern of EHSH1 in the rat CNS, using separate cultures of neurons, astrocytes, and microglia, and biochemical and immunohistochemical analyses. Cultured neurons from the cortex express primarily the long isoform EHSH1-l, as well as a small amount of the short isoform EHSH1-s. Cultured astrocytes express EHSH1-s, at a level similar to neurons, and a trace of EHSH1-l. Cultured microglia express only EHSH1-s. Double immunofluorescent staining of cortical sections showed that EHSH1 is expressed predominantly in neurons. These results suggest that EHSH1-l is a primary isoform and that EHSH1-l is highly enriched in neurons in the rat adult CNS. Immunohistochemistry of a series of brain sections revealed widespread distribution of EHSH1 throughout the brain. Particularly intense immunoreactivity was observed in the somatodendritic region of neurons in Layer III of the neocortex, hippocampus, globus pallidus, subthalamic nucleus, and substantia nigra. Interestingly, all pyramidal neurons in Layer III of the neocortex and hippocampus did not necessarily exhibit equal levels of immunostaining. In contrast, little EHSH1 immunoreactivity was detected in septofimbrial nucleus and subfornical organ of the septal region, and solitary tract and external cuneate nuclei of the medulla. Variety in the expression of EHSH1 in neurons of different regions may reflect different conditions in clathrin-mediated endocytosis and the following signal transduction. PMID- 12548703 TI - GDNF elicits distinct immediate-early gene responses in cultured cortical and mesencephalic neurons. AB - Glial cell line-derived neurotrophic factor (GDNF) has been recognized as a survival-promoting molecule for several neuronal populations in the central nervous system (CNS), including midbrain dopaminergic neurons and cortical neurons. Whereas it is well established that GDNF affects dopaminergic cell survival through a receptor complex composed of the tyrosine kinase, Ret, and the glycosylphosphatidylinositol (GPI)-anchored protein, GFRalpha-1, c-Ret is basically undetectable in cortical neurons. In the present study, we have compared GDNF signaling in cortical and mesencephalic neurons by using GDNF induced expression of the immediate-early genes, c-fos and mgif, as a readout. We found that stimulation of embryonic day (E)17 cortical cultures for 3 hr with GDNF at concentrations ranging from 10 to 80 ng/ml did not result in detectable c fos expression. In contrast, c-fos expression occurred in E14 mesencephalic cultures exposed to both low and high GDNF concentrations. Vice versa, cortical neurons responded to high GDNF concentrations (80 ng/ml) with an increase in mRNA encoding mGIF, while a similar mGIF response was absent in mesencephalic cultures. Cleavage of GFRalpha receptor subunits from their GPI anchors by phosphatidylinositol-specific phospholipase C (PIPLC) abolished GDNF-induced c fos expression in mesencephalic cultures, but did not interfere with the effects of GDNF on cortical mgif expression. Together, these findings point to distinct differences in the GDNF recognition and/or signal transduction machinery of cortical and mesencephalic neurons. PMID- 12548704 TI - Phosphodiesterase inhibitors are neuroprotective to cultured spinal motor neurons. AB - We have previously reported that cyclic guanosine-3',5'-monophosphate (cGMP) protects spinal motor neurons against acute reactive oxygen species (ROS)-induced toxicity but not against chronic ROS-induced or glutamate (Glu)-induced toxicity. In this study, we investigated the effects of phosphodiesterase (PDE) inhibitors on the survival of cultured spinal motor neurons. Selective PDE5 inhibitors (dipyridamole, T-1032, and zaprinast) as well as a nonselective PDE inhibitor (aminophylline) protected motor and nonmotor neurons against both acute ROS induced and chronic Glu-induced neurotoxicity, whereas selective inhibitors of PDE1-4 offered no protection. 8-Bromo-cGMP (8br-cGMP), a cGMP analogue, protected both motor and nonmotor neurons against acute ROS-induced toxicity but protected only nonmotor neurons against chronic Glu-induced toxicity. This neuroprotection was blocked by KT5823, a cGMP-dependent protein kinase (PKG) inhibitor. Immunohistochemical staining confirmed that PDE5 and PKG are located in almost all rat lumbar spinal neurons. Furthermore, semiquantitative analysis of the immunostaining intensity revealed that PDE5 was more abundant in motor neurons than in nonmotor neurons. Our results suggest that this difference in the amount of PDE5 may be responsible for the vulnerability of motor neurons to chronic excitotoxicity. In addition, the results of this study raise the possibility that PDE5 inhibitors might be used as a treatment for amyotrophic lateral sclerosis. PMID- 12548705 TI - Expression of delta opioid receptor mRNA and protein in the rat cerebral cortex and cerebellum is decreased by growth hormone. AB - Hormones released from the pituitary have been shown to regulate the expression of different proteins in the central nervous system. We wanted to examine whether peripheral administration of bovine growth hormone (bGH) regulates the expression of delta-opioid receptor (DOR) in the cerebral cortex and cerebellum. Expression of the DOR protein was quantified using Western blot densitometry. DOR mRNA was quantified with a solution hybridization RNase protection assay. Hypophysectomized (Hx) and untreated normal female rats were included in the study. All Hx rats were hormonally treated with cortisol (400 microg/kg/day) and L-thyroxine (10 microg/kg/day) for 19 days. Hypophysectomy resulted in a threefold increase in cerebral cortex and a twofold increase in cerebellum of the DOR protein compared with normal rats. One subgroup of Hx rats received bGH (1 mg/kg body weight) as a daily subcutaneous injection for 19 days. This treatment normalized the levels of DOR protein in the cerebral cortex and cerebellum. Immunohistochemical experiments showed that GH decreased DOR expression especially in layers II-VI in cerebral cortex and in stratum moleculare in cerebellum. Quantification of DOR mRNA by solution hybridization RNase protection assay corresponded to the DOR protein measurements. We conclude that the expression of DORs in cerebral cortex and cerebellum is regulated by GH. PMID- 12548706 TI - Inactivation of aconitase during the apoptosis of mouse cerebellar granule neurons induced by a deprivation of membrane depolarization. AB - During the excitotoxic neuronal cell death which accompanies an overflow of extracellular Ca(2+) into neurons, aconitase, an oxidative stress-sensitive enzyme of the tricarboxylic acid (TCA)-cycle in mitochondria, is inactivated due to the generation of oxidative stress (Patel et al. [1996] Neuron 16:345-355). In this study, we investigated whether aconitase could be inactivated during the apoptosis of mouse cerebellar granule cells (CGCs), which was caused by a deprivation of membrane depolarization followed by a stoppage of Ca(2+) influx into CGCs. Upon lowering the potassium (K(+)) concentration in medium from 25 to 5 mM (low K(+)), aconitase was inactivated in accordance with the decrease in methylthiazoletetrazolium (MTT)-reducing activity although its mRNA expression did not change. The blockade of Ca(2+) influx into CGCs mediated by nicardipine at 25 mM KCl also caused the inactivation of aconitase, accompanying induction of the apoptosis of CGCs. Suppression of the apoptosis of CGCs mediated by the Ca(2+) influx or neurotrophic factors such as brain-derived neurotrophic factor (BDNF) and adenylate cyclase activating polypeptide-38 (PACAP-38) attenuated the aconitase inactivation as well as the lactate dehydrogenase (LDH)-release and the decrease in MTT reduction. On the other hand, the levels of intracellular glutathione and manganese superoxide dismutase-2 mRNA decreased under the low K(+) condition, supporting a cause for oxidative stress at low K(+) due to a loss of anti-oxidant activity. Thus, the inactivation of aconitase is also caused by a deprivation of Ca(2+) influx into neurons, suggesting that aconitase is a key mitochondrial enzyme influencing the viability of neurons in response to oxidative stress. PMID- 12548707 TI - Opposite effects of alpha 1- and beta-adrenoceptor stimulation on both glutamate- and gamma-aminobutyric acid-mediated spontaneous transmission in cultured rat hippocampal neurons. AB - The effects of adrenergic receptor stimulation on spontaneous synaptic transmission were investigated in cultured rat hippocampal neurons by recording spontaneous excitatory and inhibitory postsynaptic currents (sEPSC and sIPSC). Noradrenaline (NA) inhibited sEPSC in a concentration-dependent manner, with maximal effect at 10 microM. The alpha(1)- and alpha(2)-adrenoceptor-selective agonists cirazoline and clonidine induced an inhibition of sEPSC appearance, whereas the beta-adrenoceptor agonist isoproterenol elicited an increase. The inhibitory effect of NA was reversed by alpha(1)-adrenoceptor blockade. The participation of gamma-aminobutyric acid (GABA)(B)-receptor stimulation in the inhibitory effect of NA was further examined. GABA(B)-receptor stimulation with baclofen induced a strong inhibition of bursting activity, which was fully reversed by the GABA(B) antagonist CGP 55845. By itself, CGP 55845 exerted a stimulatory effect on sEPSC frequency. In the presence of CGP 55845, the inhibitory effects of cirazoline and clonidine were maintained. NA (1, 10, and 100 microM) and alpha-adrenoceptor agonists decreased miniature EPSC and IPSC occurrence, whereas beta-adrenergic stimulation increased it. In 50% of the cells examined, NA (1, 10 microM) had a stimulatory effect on sIPSC, whereas, in the remaining 50% of cells, NA (1, 10 microM) had an inhibitory effect. In all the cells, 100 microM NA induced an inhibition of sIPSC. The inhibitory effect of NA was due to alpha(1)-receptor stimulation, whereas the excitatory effect was due to beta-receptor stimulation. In cultured hippocampal neurons, spontaneous excitatory and inhibitory synaptic transmissions are both similarly altered by adrenoceptor stimulation. However, in a subset of cells, low concentrations of NA mediate an increase of sIPSC via beta-adrenoceptor activation. PMID- 12548708 TI - Comparative genome- and proteome analysis of cerebral cortex from MK-801-treated rats. AB - cDNA microarrays and two-dimensional gel-electrophoresis in combination with mass spectrometry, were used to screen alterations in mRNA and protein levels, respectively, in cerebral cortex of MK-801-treated rats. The rats were divided in two groups; group 1 (short-term treated) and group 2 (long-term treated). In group 1, four genes were up-regulated and five down-regulated. In group 2, seven genes were up-regulated and six down-regulated. In group 1, the levels of one protein was increased and eight proteins reduced. In group 2, the levels of two proteins were increased and four proteins reduced. Several of the altered genes (casein kinase 2, glutamic acid decarboxylase, synaptotagmin, gamma aminobutyric acid [GABA] transporter, creatine kinase, and cytochrome c oxidase) and proteins (superoxide dismutase, hsp 60, hsp 72 and gamma-enolase) have previously been connected to schizophrenia. Alterations of the genes (microglobulin, c-jun proto oncogene, 40S ribosomal protein S19, adenosine diphosphate (ADP)-ribosylation factors, platelet-derived growth factor, fructose-bisphophate aldolase A, and myelin proteolipid) and the proteins (stathmin, H+-transp. Adenosine triphosphate (ATP) synthase, pyruvate dehydrogenase, beta-actin and alpha-enolase), have not, to our knowledge, earlier been implicated in schizophrenia pathology. Overall, these results with a combined approach of genomics and proteomics add to the validity of subchronic N-methyl-D-aspartate (NMDA)-receptor antagonist treatment as an animal model of schizophrenia. PMID- 12548709 TI - Gastrodin decreases immunoreactivities of gamma-aminobutyric acid shunt enzymes in the hippocampus of seizure-sensitive gerbils. AB - Gastrodin is one of the natural compound isolated from Gastrodia elata and has known anticonvulsant effects, although the exact pharmacological principles of this natural compound and its effects on other aspects of gamma-aminobutyric acid (GABA) metabolism in vivo have not been explored. Therefore, in the present study, the effects of gastrodin on GABA metabolism in the gerbil hippocampus were examined, in an effort to identify the antiepileptic characteristics of this substance. Gastrodin reduced the seizure score in the treated group, although the immunoreactivities of GABA synthetic enzymes and GABA transporters were unaltered in gastrodin-treated animals. Interestingly, in the gastrodin-treated group, GABA transaminase (GABA-T) immunoreactivity in the hippocampus, particularly in neurons, was significantly decreased. In the gastrodin-treated group, both succinic semialdehyde dehydrogenase (SSADH) and succinic semialdehyde reductase (SSAR) immunoreactivities in the hippocampus was also decreased significantly, which stood in contrast to the nontreated group, in which strong SSADH and SSAR immunoreactivities were detected. From the neuroanatomical viewpoint, these findings suggest that gastrodin may cause the elevation of GABA concentration by inhibiting the GABA shunt. PMID- 12548710 TI - Hyperbilirubinemia protects against focal ischemia in rats. AB - Heme oxygenase-1 (HO1) catalyzes oxidation of the heme molecule in concert with NADPH-cytochrome P450 reductase following the specific cleavage of heme into carbon monoxide, iron, and biliverdin, which is rapidly metabolized to bilirubin. HO1 is a stress-inducible protein that protects cells against oxidative injury, but its protective mechanism is not fully understood. The Eizai hyperbilirubinemic rat (EHBR), a mutant strain derived from the Sprague-Dawley rat (SDR), has a mutation in the gene for the canalicular multispecific organic anion transporter, which results in a phenotype of hyperbilirubinemia, and thus is a model of Dubin-Johnson syndrome in humans. In this study, we compared EHBR and SDR with regard to neuronal death induced by 2 hr of occlusion of the middle cerebral artery and reperfusion. In EHBR, the area that was immunoreactive for microtubule-associated protein-2 was significantly reduced, and the HO1 immunoreactive area was smaller than that in SDR. These results suggest that bilirubin has essentially a neuroprotective effect against focal ischemia and may participate in HO1-induced neuroprotection. PMID- 12548711 TI - Uptake of glutamate is impaired in the cortical penumbra of the rat following middle cerebral artery occlusion: an in vivo microdialysis extraction study. AB - By using microdialysis extraction of (3)H-D-aspartate and concomitant recordings of extracellular direct current (DC) potentials, the effect of middle cerebral artery occlusion (MCAO) was studied continuously over a period of 100 min in the cerebral cortex of rats. From analysis of the DC potentials, rats subjected to MCAO could be divided into three groups, one in which the dialysis probe was located in the ischemic core, one in which the probe was in the penumbra, and one in which the probe was in nonischemic tissue. In general, extraction of (3)H-D aspartate was positively correlated with the DC potential; i.e., changes in the extraction were concurrent with changes in the DC potential. Comparing the different animal groups by integration of all extraction values obtained during MCAO over time, (3)H-D-aspartate extraction was reduced by 40% in the penumbra, and by 58% in the ischemic core, compared with the sham-operated controls. No changes was found in the nonischemic group. In the penumbra group, extraction of (3)H-D-aspartate was reduced initially upon institution of MCAO but recovered to control-like levels over a period of 15-40 min, despite ongoing MCAO. In addition, extraction was reduced transiently during periinfarct depolarizations. A mean of all extraction values obtained during MCAO in the penumbra group was reduced by 47% compared with a mean of values obtained before institution of MCAO. Induction of death resulted in a reduction of (3)H-D-aspartate extraction by 86%. The present results provide direct evidence that uptake of Glu is reduced both in the ischemic core and in the penumbra of the cerebral cortex following MCAO in rats, possibly contributing to the initiation and spread of infarction. The results further indicate that uptake of Glu in the penumbra recovers to control-like levels, despite ongoing MCAO, providing evidence that Glu uptake by the Glu transporter proteins is reinstituted and/or up-regulated. PMID- 12548713 TI - Enhanced tissue plasminogen activator synthesis by the sympathetic neurons that innervate aging vessels. AB - We investigated the source of the increased release of tissue plasminogen activator (t-PA) into the circulation that occurs during natural aging. Both the basal release and the acute stress-associated release induced by sympathetic stimulations are greater in older subjects. It is widely assumed that the source of these increases is vascular endothelium. However, the sympathetic neurons that densely innervate resistance vessel walls were recently shown to synthesize and transport active t-PA to axon terminals in vascular smooth muscle, suggesting an alternative source. These fine t-PA-bearing axons lie in the seldom-studied deep adventitia of vessel walls, where they are less visible than endothelium in tissue sections. Using Northern blot analysis, we observed that t-PAmRNA synthesis is increased 54% in the ganglion parent neuron cell bodies that innervate aged vessels. The t-PA release from isolated, aged ganglia in cultures was twofold greater than that from younger controls. In addition, aged whole artery explants showed a 20% greater basal and a 50% greater acute release of stored t-PA in vitro. In vivo levels of active t-PA were 33% greater in the blood and 40% greater in the aqueous humor. These results are consistent with an increased infusion of the active t-PA protease from sympathetic axon terminals into the vessel wall extracellular matrix and the blood during natural aging, in addition to the basal endothelial release. We suggest that the cumulative impact of an accelerated plasmin production and matrix degradation within vessel walls, especially during repetitive stress, may play an unrecognized role in the pathogenesis of vascular aging. The possibility that increased sympathetic nervous system plasminogenesis influences the aging process in nonvascular tissues also deserves further investigation. PMID- 12548712 TI - Human neuronal cells (NT2-N) express functional substance P and neurokinin-1 receptor coupled to MIP-1 beta expression. AB - Substance P (SP), the most extensively studied and potent member of the tachykinin family, is a major modulator of inflammation and immunomodulatory activities within the central and peripheral nervous systems. We have examined the gene expression of SP and its receptor in a human neuronal cell line (NT2-N). Using reverse transcribed polymerase chain reaction (RT-PCR), the four isoforms of preprotachykinin-A gene transcripts (alpha, beta, gamma, and delta) were detected in the NT2-N. We also identified the presence of mRNA for neurokinin-1 receptor (NK-1R), a primary receptor for SP, in the NT2-N cells. Concomitant with NT2 cell differentiation into neurons, SP and NK-1R mRNA expression increased consistently. Intracellular SP and cell membrane NK-1R immunoreactivity were all observed in NT2-N cells. Most importantly, we demonstrated that SP and NK-1R presented in NT2-N cells are functionally involved in the regulation of macrophage inflammatory protein 1 beta (MIP-1beta), an important beta-chemokine participating in the activation and directional migration of immune cells to sites of central nervous systems (CNS) inflammation. Thus, SP and its receptor may play an important role in modulation of neuronal functions related to regulation of immune activities within the CNS. The NT2-N cell line is well suited for in vitro investigations of the SP-NK-1R pathway in immune responses and inflammation in the CNS. PMID- 12548714 TI - Vitamin D3 inhibits proinflammatory cytokines and nitric oxide production by the EOC13 microglial cell line. AB - In recent years, a neuroimmunomodulatory role for 1,25-dihydroxyvitamine D(3) [1,25(OH)(2)D(3)] has emerged. Microglial cells present a potential target for the effects of this hormone in the brain. This study focuses on the effect of 1,25(OH)(2)D(3) on the expression and production of inflammatory cytokines and nitric oxide (NO) by the EOC13 microglial cell line. The presence of the vitamin D3 receptor in microglia was demonstrated by RT-PCR. 1,25(OH)(2)D(3) inhibited the production of tumor necrosis factor-alpha, interleukin-6, and NO by stimulated microglia in a concentration-related fashion. The production of transforming growth factor-beta1 (TGF-beta1), an anti-inflammatory cytokine, was not modified in the presence of 1,25(OH)(2)D(3), indicating that the effects of 1,25(OH)(2)D(3) may not involve TGF-beta1 regulation. These results show that 1,25(OH)(2)D(3) has direct anti-inflammatory properties on microglia. It further supports the hypothesis that 1,25(OH)(2)D(3) could be involved in the maintenance of the brain homeostasis and may have a therapeutic potential in inflammatory pathologies of the central nervous system. PMID- 12548715 TI - Involvement of microglia-neuron interactions in the tumor necrosis factor-alpha release, microglial activation, and neurodegeneration induced by trimethyltin. AB - Trimethyltin (TMT) is a neurotoxicant known to induce early microglial activation. The present study was undertaken to investigate the role played by these microglial cells in the TMT-induced neurotoxicity. The effects of TMT were investigated in monolayer cultures of isolated microglia or in neuron-enriched cultures and in neuron-microglia and astrocyte-microglia cocultures. The end points used were morphological criteria; evaluation of cell death and cell proliferation; and measurements of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and nitric oxide (NO) release in culture supernatant. The results showed that, in cultures of microglia, TMT (10(-6) M) caused, after a 5 day treatment, an increased release of TNF-alpha, without affecting microglial shape or cell viability. When microglia were cocultured with astrocytes, TNF alpha release was decreased to undetectable levels. In contrast, in neuron microglia cocultures, TNF-alpha levels were found to increase at lower concentrations of TMT (i.e., 10(-8) M). Moreover, at 10(-6) M of TMT, microglia displayed further morphological activation, as suggested by process retraction and by decrease in cell size. No morphological activation was observed in cultures of isolated microglial cells and in astrocyte-microglia cocultures. With regard to neurons, 10(-6) M of TMT induced about 30% of cell death, when applied to neuron-enriched cultures, whereas close to 100% of neuronal death was observed in neuron-microglia cocultures. In conclusion, whereas astrocytes may rather dampen the microglial activation by decreasing microglial TNF-alpha production, neuronal-microglial interactions lead to enhanced microglial activation. This microglial activation, in turn, exacerbates the neurotoxic effects of TMT. TNF alpha may play a major role in such cell-cell communications. PMID- 12548716 TI - Brefeldin A-induced neurotoxicity in cultured spinal cord neurons. AB - Brefeldin A (BFA) is a fungus metabolite that is known to cause the disassembly of the Golgi complex and apoptosis in exposed cells, both of which have been suggested as playing roles in the pathogenesis of neurodegenerative diseases, particularly amyotrophic lateral sclerosis (ALS). This study showed that BFA caused neurotoxicity and apoptotic nuclear changes in cultured spinal neurons of rat spinal cord in a dose- and time-dependent manner. The spinal motor neurons were more vulnerable to this neurotoxicity. The cultured spinal neurons showed irreversible disassembly of the Golgi apparatus as early as 1 hr after exposure to BFA. BFA induced the expression and activation of caspase-12 beginning 8 hr after exposure. The level of the cleaved form of caspase-3 had increased 12 hr after the addition of BFA. Free radical generation and loss of mitochondrial membrane potential were observed in the later stages of neurotoxicity caused by BFA. Collectively, our data suggests that BFA is an excellent agent for reproducing the pathophysiological features of ALS. This in vitro model may be useful in attempts to study the mechanisms of this neurodegenerative disease and to examine therapeutic potentials. PMID- 12548717 TI - Inhibition of long-term potentiation by interleukin-8: implications for human immunodeficiency virus-1-associated dementia. AB - Human immunodeficiency virus type 1 (HIV-1)-infected mononuclear phagocytes (MP; brain macrophages and microglia) secrete a number of toxic factors that affect the pathogenesis of HIV-1-associated dementia (HAD). The identification and relative role of each MP toxin for neuronal dysfunction during HAD are not well understood. Interleukin-8 (IL-8), a CXC chemokine involved in leukocyte activation and chemotaxis, is constitutively produced by MP, and elevated levels of IL-8 mRNA were detected in the brains of patients with HIV-1 encephalitis (HIVE) by both ribonuclease protection assays and real-time PCR. To determine the role that IL-8 might play in the neuronal dysfunction in HAD, we studied its effect on synaptic transmission and plasticity in the CA1 region of hippocampus, the seat of learning and memory. Bath application of IL-8 (50 ng/ml) to rat hippocampal slices had no effect on basal synaptic transmission. However, IL-8 was shown to inhibit long-term potentiation (LTP) in a concentration-dependent manner. In control and IL-8-treated slices, the LTP magnitudes were 167.8% +/- 11.9% (mean +/- SE; n = 17) and 122.2% +/- 16.2% of basal levels (n = 13), respectively. These differences were statistically significant (P < 0.05). Preincubation of hippocampal slices with a monoclonal CXCR2 antibody (2 microg/ml) but not control IgG (2 microg/ml) blocked IL-8-induced inhibition of LTP. The expression of CXCR2 receptors in the CA1 region was shown by Western blot assays. The induction of IL-8 in HAD, its inhibition of LTP, and the expression of its receptor, CXCR2, in the hippocampus all suggest that it plays a role in the cognitive dysfunction associated with HAD. PMID- 12548718 TI - A computationally inexpensive modification of the point dipole electrostatic polarization model for molecular simulations. AB - We present an approximation, which allows reduction of computational resources needed to explicitly incorporate electrostatic polarization into molecular simulations utilizing empirical force fields. The proposed method is employed to compute three-body energies of molecular complexes with dipolar electrostatic probes, gas-phase dimerization energies, and pure liquid properties for five systems that are important in biophysical and organic simulations-water, methanol, methylamine, methanethiol, and acetamide. In all the cases, the three body energies agreed with high level ab initio data within 0.07 kcal/mol, dimerization energies-within 0.43 kcal/mol (except for the special case of the CH(3)SH), and computed heats of vaporization and densities differed from the experimental results by less than 2%. Moreover, because the presented method allows a significant reduction in computational cost, we were able to carry out the liquid-state calculations with Monte Carlo technique. Comparison with the full-scale point dipole method showed that the computational time was reduced by 3.5 to more than 20 times, depending on the system in hand and on the desired level of the full-scale model accuracy, while the difference in energetic results between the full-scale and the presented approximate model was not great in the most cases. Comparison with the nonpolarizable OPLS-AA force field for all the substances involved and with the polarizable POL3 and q90 models for water and methanol, respectively, demonstrates that the presented technique allows reduction of computational cost with no sacrifice of accuracy. We hope that the proposed method will be of benefit to research employing molecular modeling technique in the biophysical and physical organic chemistry areas. PMID- 12548719 TI - Correlation method for variance reduction of Monte Carlo integration in RS-HDMR. AB - The High Dimensional Model Representation (HDMR) technique is a procedure for efficiently representing high-dimensional functions. A practical form of the technique, RS-HDMR, is based on randomly sampling the overall function and utilizing orthonormal polynomial expansions. The determination of expansion coefficients employs Monte Carlo integration, which controls the accuracy of RS HDMR expansions. In this article, a correlation method is used to reduce the Monte Carlo integration error. The determination of the expansion coefficients becomes an iteration procedure, and the resultant RS-HDMR expansion has much better accuracy than that achieved by direct Monte Carlo integration. For an illustration in four dimensions a few hundred random samples are sufficient to construct an RS-HDMR expansion by the correlation method with an accuracy comparable to that obtained by direct Monte Carlo integration with thousands of samples. PMID- 12548720 TI - Electrostatic component of solvation: comparison of SCRF continuum models. AB - We report a systematic comparison of the electrostatic contributions to the free energy of solvation from three different kinds of quantum mechanical self consistent reaction field (SCRF) methods. We also compare the liquid-phase dipole moments as a measure of the solute's response to the reaction field of the solvent. In particular, we compare these quantities for the generalized Born model as implemented in the SM5.42R method, the multipolar expansion model developed at Nancy, and the MST version of the polarizable continuum model. All calculations are carried out at the HF/6-31G(d) level. The effects of various choices of solute cavities and representations of the charge density are examined. The test set consists of 18 molecules containing prototypical polar groups, and three different values of the dielectric permittivity are considered. PMID- 12548721 TI - Ground and excited state CASPT2 geometry optimizations of small organic molecules. AB - A method for computing second-order multiconfigurational perturbation theory (CASPT2) energy gradients numerically has been implemented and applied to a range of elementary organic chromophores, including 1,3 butadiene, acrolein, and two protonated Schiff bases. Geometries of ground and excited states-as well as conical intersections-are compared with the corresponding CASSCF structures, illustrating the effect of including the correction for dynamical electron correlation. It is shown that the differences between the two methods are not readily categorized, but that, while individual changes in bond lengths can be quite large ( approximately 0.01-0.02 A), the natures and CASPT2 energetics of the structures remain similar. Exceptions to this tend to be systems that have a strong ionic character and that are not well described at the CASSCF level. Basis set effects (double- vs. triple-zeta) were examined for a limited number of examples, and found to be quite dramatic at both levels of theory. PMID- 12548722 TI - beta-Hairpins, alpha-helices, and the intermediates among the secondary structures in the energy landscape of a peptide from a distal beta-hairpin of SH3 domain. AB - Energy landscape of a peptide, extracted from a distal beta-hairpin of src SH3 domain, in explicit water was obtained with the multicanonical molecular dynamics. A variety of beta-hairpins with various strand-strand hydrogen bonds were found in the energy landscape at 300 K. There was no energy barrier between random-coil and hairpins. Thus, the peptide conformation can easily change from the random-coil to the hairpins in the thermal fluctuations at 300 K. The landscape also included two clusters of alpha-helices, among which an energy barrier existed, and besides, these helix clusters were separated from the other conformations. Thus, the free-energy barrier exists among the helices and the other conformations. Intermediate clusters were found between the helix and the hairpin clusters. The current study showed that the isolated state of this peptide in water fluctuates among random-coil, beta-hairpin, and alpha-helix. In SH3 domain, which has a topology of mainly beta-protein, the whole-protein folding may proceed when the segment is folded in the beta-hairpin and the other parts of the protein are coupled with the beta-hairpin in an energetically or kinetically favorite way. PMID- 12548723 TI - Molecular mechanics (MM3) calculations on lithium amide compounds. AB - The MM3 force field has been extended to deal with the lithium amide molecules that are widely used as efficient catalysts for stereoselective asymmetric synthesis. The MM3 force field parameters have been determined on the basis of the ab initio MP2/6-31G* and/or DFT (B3LYP/6-31G*, B3-PW91/6-31G*) geometry optimization calculations. To evaluate the electronic interactions specific to the lithium amides derived from the diamine molecules properly, the Lewis bonding potential term for the interaction between the lithium atom and the nonbonded adjacent electronegative atom such as nitrogen was introduced into the MM3 force field. The bond dipoles were evaluated correctly from the electronic charges on the atoms calculated by fitting to the electrostatic potential at points selected. The MM3 results on the molecular structures, conformational energies, and vibrational spectra show good agreement with those from the quantum mechanical calculations. PMID- 12548724 TI - Nonspecific interaction forces at water-membrane interface by forced molecular dynamics simulations. AB - Nonspecific interactions are the main driving forces for the behavior of molecules with great affinity for biologic membranes. To investigate not only the molecular details of these interactions but to estimate their magnitude as well, the theoretical method of Forced Molecular Dynamics Simulations, based on the Atomic Force Spectroscopy experimental technique, was applied. In this approach, an additional one-dimensional elastic force, representing the cantilever probe, was incorporated to the force field of a Molecular Dynamics computational program. This force represents a spring fixed on one end to a selected atom of the molecule; the other end of the spring is displaced at constant velocity to pull the molecule out of the membrane. The force experimented by the molecule due to the spring, is proportional to the spring elongation relative to its equilibrium position. This value is registered during the entire simulation, and its maximum value will determine the molecule-membrane interaction force. Nonexplicit medium simulations were carried out. Polar and apolar media were considered according to their polarizability degree and a specific dielectric constant value was assigned. In this approach, the membrane was considered as the apolar region limited by two flat surfaces with a polar aqueous medium. The potential energy discontinuity at the interfaces was smoothed by considering the polarization-induced effects using the image method. The results of this methodology are presented using a small system, a single Alanine amino acid model, which enables extended simulations in a microsecond time scale. The confinement of this amino acid at the interface reduces its degrees of freedom and forces it to adopt one of the six defined conformations. A correlation between these stable structures at the water-membrane interface and the interaction force value was determined. PMID- 12548725 TI - Density functional investigation on electron-transfer catalysis of cycloreversion of cyclobutane: radical anion mechanism. AB - The mechanism of cycloreversion of cyclobutane radical anion (c-C(4)H(8) (-)) has been investigated at the UB3LYP/6-31++G(d,p) level, and compared with those of neutral c-C(4)H(8) and c-C(4)H(8) (+) radical cation. Although both c-C(4)H(8) ( ) and C(2)H(4) are shown to be Rydberg states unstable with respect to electron ejection, the activation barrier for the "rotating" cycloreversion of c-C(4)H(8) (-) (37.3 kcal/mol) is lower by about 25.2 kcal/mol than that of c-C(4)H(8), and even the intervention of tetramethylene radical anion intermediate may reduce the activation barrier for the cycloreversion of c-C(4)H(8) by about 8.4 kcal/mol, mainly due to stronger electron-deficiency of intermediate biradical species than close-shell cyclobutanes. For the cycloreversion for c-C(4)H(8) (-), side isomerization reaction may be efficiently prevented by the low kinetic stability of tetramethylene radical anion intermediate towards dissociation, just different from the radical cation case. Our theoretical results have suggested the possibility of electron-attachment catalysis of the cycloreversion of some electron-deficient substituted cyclobutanes. PMID- 12548726 TI - An ab initio study of intermolecular interactions of nitromethane dimer and nitromethane trimer. AB - Different geometries of nitromethane dimer and nitromethane trimer have been fully optimized employing the density functional theory B3LYP method and the 6 31++G** basis set. Three-body interaction energy has been obtained with the ab initio supermolecular approach at the levels of MP2/6-31++G**//B3LYP/6-31++G** and MP2/aug-cc-pVDZ//B3LYP/6-31++G**. The internal rotation of methyl group induced by intermolecular interaction has been observed theoretically. For the optimized structures of nitromethane dimer, the strength of C--H...O--N H-bond ranges from -9.0 to -12.4 kJ mol(-1) at the MP2/aug-cc-pVDZ//B3LYP/6-31++G** level, and the B3LYP method underestimates the interaction strength compared with the MP2 method, while MP2/6-31++G**//B3LYP/6-31++G** calculated DeltaE(C) is within 2.5 kJ mol(-1) of the corresponding value at the MP4(SDTQ)/6 31G**//B3LYP/6-31++G** level. The analytic atom-atom intermolecular potential has been successfully regressed by using the MP2/6-31++G**//B3LYP/6-31++G** calculated interaction energies of nitromethane dimer. For the optimized structures of nitromethane trimer the three-body interaction energies occupy small percentage of corresponding total binding energies, but become important for the compressed nitromethane explosive. In addition, it has been discovered that the three-body interaction energy in the cyclic nitromethane trimer is more and more negative as intermolecular distances decrease from 2.2 to 1.7 A. PMID- 12548727 TI - Boundary element solution of the linear Poisson-Boltzmann equation and a multipole method for the rapid calculation of forces on macromolecules in solution. AB - The Poisson-Boltzmann equation is widely used to describe the electrostatic potential of molecules in an ionic solution that is treated as a continuous dielectric medium. The linearized form of this equation, applicable to many biologic macromolecules, may be solved using the boundary element method. A single-layer formulation of the boundary element method, which yields simpler integral equations than the direct formulations previously discussed in the literature, is given. It is shown that the electrostatic force and torque on a molecule may be calculated using its boundary element representation and also the polarization charge for two rigid molecules may be rapidly calculated using a noniterative scheme. An algorithm based on a fast adaptive multipole method is introduced to further increase the speed of the calculation. This method is particularly suited for Brownian dynamics or molecular dynamics simulations of large molecules, in which the electrostatic forces must be calculated for many different relative positions and orientations of the molecules. It has been implemented as a set of programs in C++, which are used to study the accuracy and speed of this method for two actin monomers. PMID- 12548728 TI - Coordination number of zinc ions in the phosphotriesterase active site by molecular dynamics and quantum mechanics. AB - We have run several molecular dynamics (MD) simulations on zinc-containing phosphotriesterase (PTE) with two bound substrates, sarin and paraoxon, and with the substrate analog diethyl 4-methylbenzylphosphonate. A standard nonbonded model was employed to treat the zinc ions with the commonly used charge of +2. In all the trajectories, we observed a tightly bound water (TBW) molecule in the active site that was coordinated to the less buried zinc ion. The phosphoryl oxygen of the substrate/inhibitor was found to be coordinated to the same zinc ion so that, considering all ligands, the less buried zinc was hexa-coordinated. The hexa-coordination of this zinc ion was not seen in the deposited X-ray pdb files for PTE. Several additional MD simulations were then performed using different charges (+1, +1.5) on the zinc ions, along with ab initio and density functional theory (DFT) calculations, to evaluate the following possibilities: the crystal diffraction data were not correctly interpreted; the hexa-coordinated zinc ion in PTE is only present in solution and not in the crystal; and the hexa coordinated zinc ion in PTE is an artifact of the force field used. A charge of +1.5 leads to a coordination number (CN) of 5 on both zinc ions, which is consistent with the results from ab initio and DFT calculations and with the latest high resolution X-ray crystal structure. The commonly used charge of +2 produces a CN of 6 on the less buried zinc. The CN on the more buried zinc ion is 5 when the substrate/inhibitor is present in the simulation, and increases to 6 when the substrate/inhibitor is removed prior to the simulation. The results of both of the MD and quantum mechanical calculations lead to the conclusion that the zinc ions in the PTE active site are both penta-coordinated, and that the MD simulations performed with the charge of +2 overestimate the CN of the zinc ions in the PTE active site. The overall protein structures in the simulations remain unaffected by the change in zinc charge from +2 to +1.5. The results also suggest that the charge +1.5 is the most appropriate for the molecular dynamics simulations on zinc-containing PTE when a nonbonded model is used and no global thermodynamic conclusion is sought. We also show that the standard nonbonded model is not able to properly treat the CN and energy at the same time. A preliminary, promising charge-transfer model is discussed with the use of the zinc charge of +1.5. PMID- 12548729 TI - A practical and efficient method to calculate AIM localization and delocalization indices at post-HF levels of theory. AB - A practical and efficient method is proposed for calculating localization and delocalization indices at post-Hartree-Fock levels, and the method is tested at the CISD/6-311G++(2d, 2p) level for a large set of molecules. Our method, which utilizes wave functions written in the natural molecular orbital format and obtained with GAUSSIAN 94 or GAUSSIAN 98, convincingly extends the concepts established at the HF level. PMID- 12548730 TI - Extended method for adiabatic mode reordering. AB - The task of vibrational mode reordering is very important for reaction valley studies and for the determination of small curvature tunneling effects. An extended algorithm for adiabatic mode reordering is presented. It is based on the method introduced by Konkoli et al. [J Comput Chem 1997, 18, 1282], which is shown to suffer from numerical problems in the region of frequency-crossings and avoided crossings. One improvement is the use of cubic splines for the interpolation of the projected matrix of force constants, which allows larger step sizes between the discrete points along the reaction path, where vibrational analysis is performed. The main improvement is the use of perturbation theory to resolve crossings and avoided crossings. Within this theoretical framework it becomes clear why the method of the maximal overlap between the normal modes cannot work properly, as eigenvectors associated with nearby eigenvalues tend to become "wobbly". Thus a perturbative procedure is designed that is used for all cases where two harmonic frequencies approach each other and the overlap of the associated normal modes is of no practical use. Advantages of the new procedure are the use of larger step sizes along the minimum energy path and the much more reliable resolution of mode-crossings and avoided crossings independent of the systems symmetry. In addition to that it is shown that one should be very cautious in all computational situations when working with eigenvectors associated with nearby eigenvalues. PMID- 12548731 TI - Ab initio study on the structures of fluorinated formates and hydrogen abstraction reaction with OH radical. AB - The conformational potential energy surfaces for mono- and difluoromethyl formate have been determined by using a modified G2(MP2) level of calculations. The structures and vibrational frequencies for the conformers of mono- and difluoromethyl formate have been reported. The hydrogen abstraction reaction channels between these two formates and OH radicals have been studied at the same level of theory. Using the standard transition state theory and taking into account the effect of tunneling across the reaction barrier, we have estimated the rate constant for hydrogen abstraction by OH radical. The effect of successive fluorine substitution for methyl hydrogen on the conformational stability and on the hydrogen abstraction rate has been analyzed. PMID- 12548732 TI - DiGeorge/velocardiofacial syndrome: FISH studies of chromosomes 22q11 and 10p14, and clinical reports on the proximal 22q11 deletion. AB - DiGeorge anomaly/velocardiofacial syndrome (DG/VCFS) occurs with different deletion intervals on chromosomes 22q11, while the DiGeorge anomaly (with other findings) is seen in patients with deletions of 10p14. The clinical outcome with the common 22q11 deletion (90% of cases) is well known, but the outcome with the less frequent deletion types has not been well documented. Using cytogenetic and fluorescence in situ hybridization (FISH) analysis we studied a series of 295 patients with suspected DG/VCFS. We identified 58 subjects with a 22q11 deletion, and none with a 10p deletion. Fifty-two subjects had the common deletion, five had the proximal deletion, and one had an atypical proximal deletion due to a 1;22 translocation. We report clinical data of four subjects with the proximal 22q11 microdeletion, and of one patient with the atypical proximal deletion. The anomalies observed with the proximal 22q11 microdeletion fell within the DG/VCFS spectrum. Two females, 6 and 25 years old, had normal mental development. Normal development has been reported with the common 22q11 deletion, but only in a minority of cases. This study may indicate a better intellectual and/or behavioral outcome with the proximal vs. the common 22q11 deletion, rather than a chance finding. PMID- 12548733 TI - Female carriers of fragile X premutations have no increased risk for additional diseases other than premature ovarian failure. AB - Carriers of fragile X premutations were previously considered phenotypically normal but are now known to be at risk for premature ovarian failure (POF). This prompted us to investigate whether premutation carriers (PC) have an increased risk for other diseases. We approached 306 women out of 84 fra(X) families of whom 264 (86.3%) participated in this study. A medical history was taken of these women. Whenever possible, the anamnestic data were verified with medical records. We first evaluated the occurrence of diseases that are commonly associated with menopause in PC and compared this to that in women with either a normal FMR-1 gene or a full mutation. We found no statistically significant differences in the occurrence of diseases known to be associated with menopause, such as cardiovascular diseases and osteoporosis. However, lower bone mineral density was observed only in PC. Subsequently, we compared the occurrence of other medical problems between the two groups by estimating relative risks. PC did not demonstrate other diseases more commonly compared to non-PC from the same families. These findings are important for the counseling of PC. Carriership of the premutation may affect the ovaries, but does not substantially increase the risk for additional medical problems. Once a PC does experience POF, she is at risk for early estrogen deprivation, which may lead to a premature decrease in bone density, when not treated. PMID- 12548734 TI - Extended phenotype of pontocerebellar hypoplasia with infantile spinal muscular atrophy. AB - Pontocerebellar hypoplasia (PCH) is rarely associated with anterior horn cell disease and designated as PCH-1. This phenotype is characterized by severe muscle weakness and hypotonia starting prenatally or at birth with a life span not exceeding a few months in most cases. Milder disease courses with later onset and longer survival are normally not diagnosed as PCH-1. We describe the clinical and neuroradiological findings in nine patients out of six siblingships with evidence of cerebellar defects and early onset spinal muscular atrophy (SMA), representing a broad spectrum of clinical variability. In all patients, the diagnosis of SMA (Werdnig-Hoffmann disease) was made on the basis of electrophysiological data and muscle biopsy; however, genetic testing failed to confirm the diagnosis of infantile SMA with a gene defect on chromosome 5q and resulted in clinical reevaluation. Age at onset was after a normal period in the first months of life in three siblingships and pre- and postnatally in the other three families. Life span was 2-4 years in patients with later onset, and age at death occurred after birth or within months in the more severe group. Two siblingships showed discordant ages at death despite similar treatment. In contrast to the previous definition of PCH-1, our observations suggest the existence of milder phenotypes with pontocerebellar hypoplasia or olivopontocerebellar atrophy in combination with anterior horn cell loss. A pontine involvement is not necessarily seen by neuroimaging methods. The genetic basis of PCH-1 remains to be determined. The gene locus for infantile SMA on chromosome 5q could be excluded by linkage studies. Parental consanguinity and affected siblings make autosomal recessive inheritance most likely. PMID- 12548735 TI - Exclusion of RNX as a major gene in congenital central hypoventilation syndrome (CCHS, Ondine's curse). AB - Congenital central hypoventilation syndrome (CCHS) is a rare condition for which segregation analyses have suggested genetic factors. The respiratory phenotype of Rnx knock-out mice together with the Rnx expression at the brainstem level prompted us to consider the RNX gene as a candidate for CCHS in human. The screening of the RNX gene in a series of 25 patients with CCHS did not reveal any significant nucleotide variation. We therefore conclude that RNX is not a major gene for CCHS in human. PMID- 12548736 TI - Biological and environmental contributions to adaptive behavior in fragile X syndrome. AB - Fragile X syndrome is the most common cause of inherited developmental disability. The purpose of the present study is to understand how both biological and environmental influences affect the development of adaptive behavior in children with fragile X. In-home assessments were conducted on 120 children (80 boys and 40 girls) with the fragile X full mutation and their unaffected siblings (58 boys and 62 girls). Hierarchical multiple regression analyses were utilized. Independent variables included biological or demographic variables specific to the child (age, gender, full-scale IQ, and FMRP percentage), as well as factors specific to his/her environment (household income, home environment, maternal psychopathology, and effectiveness of educational/therapeutic services). Results of these analyses showed that for both boys with fragile X and the control sibling group, adaptive behavior was predicted by IQ, age, gender, and home environment. For girls with fragile X, adaptive behavior was most strongly associated with IQ. Adaptive behavior was not significantly associated with FMRP in boys or girls with fragile X. By examining the relative influences of biological and environmental factors on adaptive behavior in children with fragile X, we begin to lay the foundation for the development of more specific treatment studies in children with this disorder. PMID- 12548737 TI - Chromosomal fragility in patients with triple A syndrome. AB - Triple A syndrome is a rare, autosomal recessive disorder characterized by alacrima, achalasia, and adrenal insufficiency. Previous studies have shown that the triple A gene (AAAS) maps to chromosomal band 12q13. Mutations in the AAAS gene have been identified in triple A syndrome patients; however, the function of this gene is still obscure. We used classical and high-resolution chromosome analyses along with chromosome painting and DNA sequencing to study patients with triple A syndrome. We observed abnormalities in the heterochromatic region of chromosome 9 that included chromatid breaks, chromosome breaks, whole chromosome arm loss, and marker chromosomes, which occurred at unusually high frequencies in affected patients and heterozygotes. Our study raises the possibility of an association between chromosomal fragility in band 9q12 and triple A syndrome. Further investigation is necessary to understand the biologic basis of this finding in the context of triple A syndrome. PMID- 12548738 TI - A new autosomal recessive congenital contractural syndrome in an Israeli Bedouin kindred. AB - We describe 23 cases with a syndrome of congenital contractures belonging to a large, inbred Israeli-Bedouin kindred. The phenotype described is similar to the Finnish type lethal congenital contracture syndrome yet differs in the following ways: by some additional craniofacial/ocular findings, by the lack of hydrops, multiple pterygia, and fractures, and by the normal duration of pregnancy. The major unique and previously undescribed clinical feature in our patients is a markedly distended urinary bladder as well as other urinary abnormalities. The vast majority of the cases died shortly after birth. Sonographic prenatal diagnosis was possible as early as 15 weeks gestation by demonstrating fetal akinesia, limb contractures, hydramnios, and distended urinary bladder. Linkage to 5q and 9q34 loci has been excluded. PMID- 12548739 TI - Disclosing the mechanisms of origin of de novo short-arm duplications of chromosome 9. AB - Three de novo short-arm duplications of chromosome 9 were investigated by fluorescence in situ hybridization (FISH) and genotyping of microsatellite loci with the aim of disclosing their mechanisms of origin. Two of these duplications were identified as direct and one as an inverted duplication, and they comprised nearly the entire 9p. In the two dirdup(9p), the distal breakpoints were mapped to subtelomeric sequences, whereas the proximal break boundaries were defined by pericentromeric sequences. The maternal origin of both duplications was demonstrated, and the presence of three different alleles in distal loci indicated that these rearrangements must have occurred prezygotically. In the invdup(9p), the proximal breakpoint was mapped at the alphoid sequences, which were partially duplicated on the tip of the rearranged 9p; subtelomeric and telomeric sequences were not detected, allowing the distal breakpoint to be mapped proximally to subtelomeric sequences. Genotyping of microsatellite loci allowed us to conclude that this invdup(9p) chromosome had a paternal origin. Homozygosity throughout the duplicated segment pointed to its probable postzygotic formation. Chromatid rearrangements appeared to have originated these 9p duplications, involving pericentromeric and subtelomeric sequences. These rearrangements might have been the result of recombination events between homologous repeats in these segments. PMID- 12548740 TI - Facial dysgenesis: a novel facial syndrome with chromosome 7 deletion p15.1-21.1. AB - We describe a female neonate with a unique constellation of features including anophthalmia and cryptophthalmos, temporal remnant "eye tags," bilateral cleft lip, unilateral cleft palate, a proboscis with absent nasal septum, choanal atresia, micrognathia, square stoma, and bilateral external auditory canal atresia. Gross brain structure, pituitary function, limbs, trunk, and genitalia were normal. Skeletal survey, echocardiogram and abdominal viscera were unremarkable except for a split central sinus of the right kidney. BAER exam indicated she could hear and temporal CT confirmed the presence of cochlea and possible ossicles. Cytogenetic evaluation revealed an interstitial deletion at chromosome 7p15.1-21.1. TWIST, a gene encoding a transcription factor involved in craniofacial development, is deleted by FISH analysis. The absence of a mutation on the non-deleted allele of TWIST as determined by sequencing virtually eliminates complete loss of the TWIST gene as the cause of this patient's severe phenotype. The HOXA gene cluster also encodes transcription factors that are crucial for directing cephalad to caudad somatic fetal development. HOXA1, the most telomeric of the 13 members of the HOXA gene cluster, is located at the centromeric boundary of the patient's chromosome 7 deletion. By FISH analysis, neither allele of HOXA1 is deleted and sequencing reveals no mutations. Haploinsufficiency or complete loss of the HOXA1 gene also does not appear to cause this patient's severe phenotype. Previous reports of chromosome 7p15-21 deletions do not have phenotypes similar to this patient. PMID- 12548741 TI - Recombinants of intrachromosomal transposition of subtelomeres in chromosomes 1 and 2: a cause of minute terminal chromosomal imbalances. AB - Two cases of submicroscopic recombinants of intrachromosomal transposition of telomeres, one each from chromosome 1 and 2 are described. Meiotic crossing-over would generate the recombinants from these reciprocal rearrangements. In both cases, which were detected by FISH with subtelomeric probes, there is a minute deletion of the qter region and a second presence of the pter subtelomeric region on the respective qter, i.e., a duplication of 1pter or 2pter respectively. The deletion on 2qter (case 2) was confirmed by microsatellite inheritance and was of paternal origin, but in case 1 there was no detectable 1q deletion other than of the subtelomeric probe, and parental origin could not be determined. The present case 2 with del(2qter)/dup(2pter) shares many features with reported cases of simple deletion (2qter) but did not have features of Albright hereditary osteodystrophy, which are seen in half of such deletion patients. The clinical features present in case 1 were similar to those of the previously reported case of a submicroscopic 1qter deletion but also to cases with microscopically visible 1qter deletions, presumably because of gene enrichment in subtelomeric regions. Recombinants of such intrachromosomal subtelomere transpositions detected by subtelomeric probes may comprise up to 10% of submicroscopic pter or qter deletion cases. Other cases of this unusual mechanism may be detected with more common use of subtelomeric probes. It is suggested the bouquet associations of telomeres in early meiosis may facilitate such unusual rearrangements. PMID- 12548742 TI - Cryptic t(1;12)(q44;p13.3) translocation in a previously described syndrome with polymicrogyria, segregating as an apparently X-linked trait. AB - We report on the multistep progression to the correct genetic diagnosis in an apparently new syndrome of mental retardation and multiple congenital anomalies, including hypogenitalism and polymicrogyria. We had previously reported it as an X-linked condition affecting four members (three males and one female) of a family [Zollino et al., 1992: Am J Med Genet 43:452-457]. Two of the four patients, both males, presented with a brain abnormality that was initially described as pachygyria, while the remaining two (one male and one female) did not. Our present study includes a clinical follow-up on the patients, neuroradiological reexamination of one patient, X linkage studies and X inactivation analyses, and finally molecular cytogenetics, which allowed us to establish definitely the genetic causes of the condition. After the detection of a subtle t(1;12)(q44;p13.3) balanced translocation in healthy carriers, two unbalanced segregation products were observed in different patients, resulting in 1q44qter monosomy and 12p13.3pter trisomy in patients with polymicrogyria and severe psychomotor delay, 12p13.3pter monosomy and 1q44qter trisomy in the other two patients without polymicrogyria, with less severe mental retardation and less distinctive physical anomalies. Thus, this condition is no longer to be considered X-linked, but the result of cryptic autosomal imbalance. Furthermore, this study identified an approximately 14 Mb interval in 1q44qter pathogenetically related to polymicrogyria. PMID- 12548743 TI - Bilateral microphthalmia with cyst, facial clefts, and limb anomalies: a new syndrome with features of Waardenburg syndrome, cerebro-oculo-nasal syndrome, and craniotelencephalic dysplasia. AB - We report a patient with bilateral microphthalmia with cyst, limb anomalies, and multiple facial malformations. This patient has clinical features similar to Waardenburg ophthalmo-acromelic syndrome, cerebro-oculo-nasal syndrome, and craniotelencephalic dysplasia. Although all of these syndromes are characterized by microphthalmia, the presently reported patient does not have the complete pattern of any of these syndromes, It is possible that he has a previously undescribed syndrome, most closely related to the cerebro-oculo-nasal syndrome with malformations outside the craniofacial region. More case reports are needed to further delineate this possibly new syndrome. PMID- 12548744 TI - De novo interstitial tandem duplication of chromosome 20p12.1p13. AB - We report the first case of an individual with a de novo interstitial tandem duplication of the short arm of chromosome 20p12.1p13, discuss the clinical features, and propose the possible underlying mechanism of formation. PMID- 12548745 TI - Pachygyria and polymicrogyria in 22q11 deletion syndrome. PMID- 12548746 TI - Aural atresia, microtia, complex heart defect, and hearing loss syndrome: additional case. PMID- 12548747 TI - Prenatal diagnosis of a rare chromosomal instability syndrome: variegated aneuploidy related to premature centromere division (PCD). PMID- 12548748 TI - Is Sonic hedgehog (SHH) a candidate gene for spina bifida? A pilot study. PMID- 12548749 TI - De novo mutation in the gene encoding connexin-26 (GJB2) in a sporadic case of keratitis-ichthyosis-deafness (KID) syndrome. PMID- 12548750 TI - Autopsy on a case of Roberts syndrome reported in 1672: the earliest description? AB - In 1672, Francois Bouchard reported the autopsy of a male infant discovered on public exhibition at Leiden. The findings of tetraphocomelia more severe in the upper limbs with reduction in the number and length of digits, hydrocephalus, bilateral cleft lip, micrognathia, and cryptorchidism are consistent with a diagnosis of Roberts syndrome. Bouchard did not find internal abnormalities other than hydrocephalus, which he attempted to demonstrate by showing decreased specific gravity of the brain. Death was due to umbilical vein hemorrhage following birth trauma. PMID- 12548751 TI - Clinical paper and statistics. PMID- 12548752 TI - Reply to correspondence from Axenovich and Borodin--"Some pitfalls of segregation analysis of complex traits". PMID- 12548753 TI - [Some differences on biological characteristics of Campylobacter jejuni]. AB - Fecal samples from 195 diarrheic patients in different age, rectal and cloacal swabs from 656 normal and diarrheic animals and poultry, 108 visceral materials from the dead animals with diarrhoea were cultured for Compylobacter jejuni. A total of 458 strains of campylobacters (445 strains of C. jejuni, 13 strains of C. coli) were isolated and identified, and some biological characteristics of these strains were observed. Lior's biotyping scheme was used for biotyping 354 strains of campylobacters(253 strains of C. jejuni and 2 strains of C. coli). The results showed the most biological characteristics of these campylobacters accorded with that previously described in the literatures, but it were also found that there were some differences on morphological, cultural, physiological and biochemical characteristics and antibiotic resistances. In these differences, the most main differences were that 48.3% (215/445) of C. jejuni and 23.1% (3/13) of C. coli were resistant to nalidixic acid, and that 1.1% (5/445) of C. jejuni and 7.6(1/13) of C. coli were resistant to cefotaxime. There were relationships between antibiotic resistances and strain sources (P < 0.005). The result of biotyping 352 strains of C. jejuni indicated that biotype I (40.9%) and II (58.2%) were predominant in the bodies of these animals, and there were two biotypes distributed in the body of same animal. PMID- 12548754 TI - [Studies on application for classification and identification in Mycobacterium by analysis of PCR amplification of 16S-23S ribosomal DNA spacer sequences]. AB - The 16S-23S ribosomal DNA spacer sequence of mycobacterium were amplified by PCR. The products were visualized by PAGE, and evaluate the possibility for classification and identification of mycobacterium at gene level. The sensitivity of PCR in annealing temperature 45 degrees C was 500 fg/microL, whereas 50 degrees C was 5 pg/microL. The results showed that: the amplified bands ranging from 300 to 600 bp, most of rapid-growing Mycobacterium and Nonmycobacterium tested have more bands and the bands molecular weights were larger than slow growing Mycobacterium. The relativity of mycobacterium < 70%, most of them < 50%. This experimental method might be rapid and effective for differentiation of Mycobacterium at species level. PMID- 12548755 TI - [Construction of genomic library from L. lactis AL2 and isolation of entire nisin biosynthesis gene cluster]. AB - A total DNA library of Lactococcus lactis AL2 with high yield of nisin was successfully constructed using lambda EMBL3 as vector. 3900 plaques were obtained, which is much more than the required number of recombinants that represents an entire L. lactis genome according to Clarke and Carbon formula. Southern hybridization, PCR amplification and DNA sequencing revealed that the entire nisin biosynthesis gene cluster was isolated from the constructed library. PMID- 12548756 TI - [Cloning and expression of otsA gene in E. coli]. AB - 1.5 kb of otsA gene encoding trehalose synthase has been cloned by PCR amplification. The DNA fragment was ligated to multi-copy vector and transformed to otsBA deleted and otsA deficient strains of E. coli separately. The transformants exhibited growth as well as the otsBA+ wild type on medium containing 0.5 mol/L NaCl. Trehalose was synthesized and accumulated in the transformed cells under osmotic pressure, which was determined by thin layer chromatograph. The results confirmed that otsA gene was functionally expressed in the recipient strains. These studies suggested that engineering otsA gene and trehalose accumulation into crop plants may improve drought and salinity tolerance. PMID- 12548757 TI - [Studies on gene knocking out of 2-keto aldose reductases from Erwinia sp. SCB125]. AB - Based on the reported gene sequences, the segments containing 2-keto aldose reductase (2-KRA and B) genes were amplified by PCR from the plasmids and Erwinia sp. SCB125 each for gene expression and gene knocking out. Then cloning them into expression vector pBL and successfully expressing them with high enzyme activity in E. coli DH5 alpha. After their enzyme activities were proved, the work on gene knock out followed. Introducing the knock-out vector which distribute unstably during the cell division to the host strains Erwinia sp. SCB125. Screening firstly by the positive marker, one resistance which resulted from the expression of the resistance gene inserting inside the reductase genes and the negative marker, another resistance which outside the reductase genes in the vector. The strains selected out will be tested by further study. This work was the bases of blocking the pathway metabolism and constructing a recombinant strain that can produce 2-KLG directly from D-glucose by one-step fermentation. PMID- 12548758 TI - [In vitro antiviral activity of antisense oligonucleotides against influenza virus]. AB - For developing of antisense oligonucleotides as potential antiviral therapeutic agents against influenza A virus, phosphorothioate oligodeoxynucleotides(PS-ODN) targeted to part to the 3' and 5' end sequences which are common to the eight RNAs of type A influenza virus were synthesized. The in vitro cytotoxicity of these PS-ODNs was assayed and then antiviral activity of these PS-ODNs was evaluated by using cultured MDCK cells infected by A/JingFang/86-1(H1N1). It was found that(1) No in vitro cytotoxicity was seen when concentration of PS-ODN was up to 50 mumol/L. (2) Oligonucleotides(IV4 #) complementary to 5' terminal conserved sequences and ODN(IV6 #, IV7 #)composed by 3'/5' terminal conserved sense sequences or antisense sequences all exhibited the most potent antiviral activity. IV4 # ODN exhibited significant antiviral activity with reduced hemegglutination titer about 50% at concentration of 1 mumol/L, and over 70% at 10 mumol/L. The antiviral activity of IV4 # was in a dose- and sequence-dependent manner. (3) IV4 # ODN showed significant antiviral activity not only against H1N1 subtype strains of influenza virus, but also against H3N2 subtype strains. (4) The multiplicity of infection(MOI) of influenza virus had effects on antiviral activity of IV4 # ODN. IV4 # ODN showed better dose-dependent antiviral activity when there was lower MOI. These results suggested the the 3'/5' terminal sequences of influenza A virus could be used as a target for designing antiviral oligonucleotides against influenza A virus. PMID- 12548759 TI - [cDNA cloning and sequence analysis of the seventh segment of maize rough dwarf virus genome]. AB - The double strand RNA of maize rough dwarf virus (MRDV) was prepared from the maize samples showing symptoms which was from the Luanchen county of Heibei province of China. The primers were designed according to the known sequence of MRDV, the cDNA sequence of the seventh segment of MRDV was obtained by RT-PCR, the S7 sequence was analyzed by computer after sequencing. The results showed: the full length of the S7 cDNA is 1936 bp and equal to that of the S7 cDNA from abroad, the two open reading frame(ORF1 and ORF2) contained in the S7 segment are also unchanged. In comparison with the S7 segment from Italy, the homology of S7 nucleotide is 87.7% and the homology of ORF1 amino acid sequence is 91.6%. However, the MRDV S7 segment and the rice black strike dwarf virus S8 segment showed 95.5% nucleotide identities and 93.5% ORF1 amino acid identities. PMID- 12548760 TI - [Immune responses of a recombined live Salmonella typhimurium SL3261 expressing a multi-epitope antigen of HCV]. AB - A multi-epitope antigen gene of hepatitis C virus(HCV) was fused to beta galactosidase gene and introduced into attenuated Salmonella typhimurium SL3261 to construct HCV recombined live vaccine candidate SL3261 (pWR/PCX). when the oral live bacteria were used to immunize mice or rabbits, specific anti-GZ-PCX IgG was detected at week 6 and the strongest antibody responses happened at week 12 at a titer of 1:800 and 1:25,600 in mice and rabbits, respectively, which showed significant difference compared with those of SL3261 and blank controls. Anti-GZ-PCX sIgA in mice's intestine and anti-LPS antibody in sera were also detected. The oral live bacteria elicited obvious DTH reaction and proliferation response of peripheral lymphocytes by GZ-PCX antigen. The body weight of immunized mice slightly decreased but no other toxic effects was observed, which showed the safety of oral immunization. The study of oral live HCV multi-epitope vaccine might be able to provide a new route for the researches of HCV vaccines. PMID- 12548761 TI - [Studies on the correlation between production of L-malic acid and some cytosolic enzymes in the L-malic acid producing strain Aspergillus sp. N1-14]. AB - The cytosol enzymatic study in the case of high L-malic acid(LMA) production of Aspergillus sp. N1-14' was reported. The activities of 4 kind enzymes that catalyse the CO2 fixation reactions have been detected, which are pyruvate carboxylase(PC), phosphoenolpyruvate carboxlase (PEPC), phosphoenolpyurvate carboxykinase(PCK) and malic enzyme(ME). With the exception of ME, the linear correlation was found between activities of three carboxlases and the production rate of LMA. The activity of malate dehydrogenase(MDH) was at the level of 2-3 exponential higher than that of the other analysed enzymes, while the activity of succinate dehydrogenase(SDH) was much lower, and as a discrepancy, SDH was in a positive correlation to the content of LMA in fermenting slurry(r = 0.9252). It is shown that the accumulated LMA acted as an activator of SDH. Through dynamic study, it is found that, in contrast with the slow and even increase of biomass, the content of cytosol protein(Cp) sharply fluctuated mainly due to the changes of aeration conditions. The data of the linear correlation coefficients(r) of activities of cytosol enzymes to Cp(PC r = 0.9563, PEPC r = 0.7688, PCK r = 0.7300, MDH r = 0.3920, SDH r = -0.2086) exhibited an inner law of protein synthesis. Experiment of increasing the amount of spore inoculum resulted in increase of LMA and decrease of SA. After fermenting 120 h in a 5 L stirred fermentor, with 3-fold of original spore inoculum 105.88 g/L of LMA was achieved, the overall productivity was 0.883 g/(L.h), the converting rate of glucose to LMA was 78.43%. This result supports the exist of a inner law of protein synthesis in the early period of LMA fermentation by Aspergillus sp. N1-14'. PMID- 12548762 TI - [Cultivation of Lactobacillus in microcapsule]. AB - The NaCS-PDMDAAC capsules were used to immobilize the two kinds of lactobacillus. The experiment results showed that two kinds of lactobacillus could grow well, and the densities of cells were 1.8 x 10(11)/mL capsule and 2.79 x 10(11)/mL capsule, respectively which was much higher than that in free cell culture. The conversion rate of lactic acid could be 2-5 folds higher than that in free cell culture. PMID- 12548763 TI - [Isolation of moniliformin-degrading bacterium Ochrobactrum sp. and analysis of its functional properties]. AB - A moniliformin(MON)-degrading bacterium strain, named as Y21-2, was isolated from the mycotoxin-contaminated soil from Heilongjiang Province by the enrichment microculture technique. This strain can grow with MON as its sole carbon and energy source. In the minimal medium with 500 micrograms/mL MON, the number of cells increased from 10(7) to 10(10). Traditional taxonomy, assays of its G + C content and 16S rDNA sequence homology identified Y21-2 as Rhizobiaceae, Ochrobactrum sp. Resting cell suspensions prepared from induced Y21-2 can degrade MON with great speed, which also suggested the existence of enzymes committed to MON degradation in the cell. PMID- 12548764 TI - [Effect of temperature on the activity of some enzymes representative of EMP pathway and TCA cycle in psychrophilic yeast]. AB - Some key enzymes of EMP pathway and TCA cycle in a psychrophilic yeast Y18 were studied in this paper compared with those of Saccharomyces cerevisiae. The results indicated that fructose, 1,6-bisphosphate aldolase, succinate dehydrogenase, and hexokinase in Y18 were very thermolabile and have high activity at low temperature. These enzymes belong to cold-active enzymes. Alpha ketoglutarate dehydrogenase existed possibly in isoenzyme which had different temperature characteristics. Citrayl synthetase was very similar in temperature characteristics to that of mesophiles. The Km value of succinate dehydrogenase both from Y18 and S. cerevisiae were studied and Some features of enzyme in psychrophiles were also discussed in this paper. PMID- 12548765 TI - [Production, partial purification and characterization of methamidophos-degrading enzyme from methylotroph WB-1]. AB - After cultivation for 20 h in the inorganic salt medium with methamidophos as sole carbon and nitrogen source, Methylotroph WB-1 could produce methamidophos degrading enzyme in larger amounts. The enzyme was partially purified by sonication disruption, Tween-20 extraction, heat treatment(9 min at 60 degrees C), DEAE-cellulose 32 and CM-cellulose32 chromatography with 22.8 times purification and 78.8% recovery. Activity staining showed single violate band corresponding to that of protein staining. Optimum pH of the enzyme was 9.0; poor substrate specificity was showed. It was strongly inhibited by Hg2+, Mn2+ and Cu2+, but not by Co2+ and Zn2+; it also exhibited poor stability. PMID- 12548766 TI - [The important role of vitamins in the over-production of pyruvic acid]. AB - The effect of nicotinic acid, thiamine, pyridoxine, biotin and riboflavin on the production of pyruvic acid by Torulopsis glabrata WSH-IP303 with glucose as carbon source and NH4Cl as sole nitrogen source was investigated. By using orthogonal experiment method, thiamine was confirmed to be the most important factor affecting the production of pyruvic acid. Based on a certain concentration range of thiamine (0.01-0.015 mg/L), glucose consumption rate can be enhanced by increasing the concentration of nicotinic acid. When the concentration of nicotinic acid, thiamine, pyridoxine, biotin and riboflavin were 8, 0.015, 0.4, 0.04 and 0.1 mg/L, respectively, the concentration and yield to glucose of pyruvic acid reached 52.4 g/L and 0.525 g/g at 48 h in flask culture, respectively. Batch culture was conducted in a 2.5 L fermentor with initial glucose concentration of 120 g/L. By adopting the optimal concentration combination of vitamins, the concentration and yield to glucose of pyruvic acid reached 69.4 g/L and 0.593 g/g at 57.5 h, which were increased by 32.4% and 13% than the best results in flask culture, respectively. PMID- 12548767 TI - [Studies on biosorption of Pd2+ by bacteria]. AB - The strain R08, showed relatively strong ability of adsorbing Pd2+, was screened from different source of bacterial strains. The strain R08 was identified as Bacillus licheniformis. The optimum pH value of Pd2+ biosorption by R08 dead biomass is 3.5. The biosorption is a rapid and non-temperature dependent process. The biomass and Pd2+ initial concentration in solution affected the biosorption process. The bosorptive capacity could reach 224.8 mg/g under the conditions of Pd2+ 200 mg/L, 0.4 g biomass/L, pH3.5 and 30 degrees C for 45 min. TEM observation showed that R08 biomass is able to adsorb and reduce Pd2+ ions to Pd0 particles. IR spectrum analysis indicated that -COO- and HPO(4)2- groups on the cell walls may involve in process of adsorbing Pd2+. PMID- 12548768 TI - [Study on the biology feature and pathogenicities to silkworm of a microsporidium isolated from Barathra brassicae L]. AB - A microsporidium (called as Bab-M) was isolated from Barathra brassicae L. captured from suburban vegetable plot of Guang Zhou. The spores were long-ovoid in shape and 4.02 +/- 0.36 microns x 1.99 +/- 0.36 microns in size. Immunologically the microsporidium shared spore surface specific antigen(s) with N. bombycis. The ultrastructure and life cycle of Bab-M were similar to that of N. b.. The rate of transovarian transmission was high. The initial conclusion was that Bab-M should be referred to as Nosema bombycis, but there was variation between them. PMID- 12548769 TI - [Genetical analysis on cross incubation of Beauveria bassiana]. AB - Heterokaryon of Beauveria bassiana was formed during the cross incubation of two vegetative compatible strains with genetic markers of actidione resistance and 34 degrees C tolerance. The chromosome(s) or its fragment successive losses, recombination and segreation led to haploidization during the conidia formation period. After at least 4 generations of parasexual cycle, the genetic character of heterokayon could get to relative stable. Genetic marker and RAPD analysis indicated that the combinants showed the phenomenon of preferential selection of one parental type by unrandom chromosome lossing and the gene of the other parental strain was suppressed or lost completely. Different culture medium, in vivo or in vitro, and different mixture ratio of original strain spores could affect parasexual process and then the preferential selection. The results also demonstrated the heterosis effect of cross culture. PMID- 12548770 TI - [Tn5 mutagenesis and the characteristics of indole-3-acetic acid biosynthesis in Alcaligenes faecalis A1501]. AB - The IAA production of Alcaligenes faecalis wild type strain A1501 depended on addition of L-tryptopham(trp). A1501 could grow well in minimal medium without Trp, but could not produce IAA. The results indicated that there was Trp dependent IAA biosynthesis pathway in A. faecalis A1501. A1501 shows a characteristic of cell density-dependent IAA biosynthesis. The Tn5-induced mutant bank of A1501 was constructed using Tn5 mutagenesis and one Trp auxotrophic mutant AT63 was selected and identified. The Tn5-induced mutant AT63 could synthesis 2.24 micrograms/mL IAA in minimal medium without L-Trp. It indicated there were at least two IAA biosynthesis pathways in A. faecalis, one needed Trp, the other needed IGP as precursor. The results of southern hybridization also suggested that the Tn5-insertion might locate in gene locus encoded IAA synthetase. PMID- 12548771 TI - [Precipitation of chlorella virus FJ-1 by polyethylene glycol(PEG8000)]. AB - Chlorella virus FJ-1 was isolated in Fujian Province of China. It can be precipitated by polyethylene glycol(PEG8000) and the best condition if 7% PEG8000 and 4% NaCl. But the precipitation efficiency of PEG8000 is lower than that of ultracentrifugation. PMID- 12548772 TI - [Isolation and identification of a killing maggots bacterium]. AB - A notably killing maggots bacterium was isolated from natural dead maggots in the manure pits in the countryside of Yancheng. Its pathogenicity was confirmed by the law of KOCK. The results of preliminary bioassay show that the pathogen can infect the larvas of greenbottle flies and other larvas of flies in a certain extent, but can't infect animals and fowls. The G + C content of its DNA is 62.46%. The hybridization ratio of its DNA and the Pseudomonas pseudoalcaligenes' (AS1.1806) is 81.2%. According to Bergey's Manual of Systematic Bacteriology Ninth edition, the strain of the bacterium was primarily identified as Pseudomonas pseudoalcaligenes. PMID- 12548773 TI - [Fe(2+)-oxidation and sulphur-oxidation system of Thiobacills ferrooxidans and its molecular genetics]. PMID- 12548774 TI - [Studies on microinjection of mouse spermatozoa into rabbit oocytes]. AB - The methods of intracytoplasmic sperm injection (ICSI) and subzonal injection (SUZI) were used to study heterologous fertilization and embryonic development between the mouse and the rabbit. Results were as follows: 1. The mouse sperm nuclei decondensed and formed pronuclei following microinjection into cytoplasm and perivitelline space (PVS) of rabbit oocytes; 2. The hybrid embryos developed to the stage of 8-cell when cultured in vitro; 3. The karyotype analysis showed a normal complement of rabbit oocyte and mouse sperm chromosomes in the 4-cell hybrid embryos; 4. The ultrastructure of 4-cell hybrid embryos was similar to that of normal 4-cell rabbit embryos; 5. The fertilization rate (32.4%) and cleavage rate (22.2%) when 5-10 mouse spermatozoa were injected were higher than those of injection of a single spermatozoon into PVS of the rabbit oocyte, but the difference was not significant (P > 0.05). The fertilization rate (42.3%) and cleavage rate (30.8%) in rabbit oocytes in vitro matured for 11-12 h were higher than those in the oocytes which were in vitro matured for 24-25 h following microinjection of 1-2 mouse spermatozoa into PVS, but the difference was not significant (P > 0.05). PMID- 12548775 TI - [Distribution of luteinizing hormone (LH) and human chorionic gonadotropin (hCG) in the nervous system, Hatschek's pit and gonads of protochordata]. AB - Immunocytochemical localization of the nervous system, Hatschek's pit and gonads in protochordata was carried out using two specific antibodies against human LH and hCG. The results indicate that there are two different kinds of LH- and hCG like immunopositive cells in the brain vesicle and nerve tuber of amphioxus, and nerve ganglion in Styela plicata showed immuno-negative reaction to LH and hCG antibodies. At the same time, we found for the first time that LH- and hCG-like immuno-positive cells existed in the early gonads (ovaries and testis) in protochordata. Positive substance distributed in the cytoplasm, nucleoplasm and nucleolar membrane of oogonia as well as early spermatogenic cells in testis. Afterwards, this positive substance was still distributed in the cytoplasmic membrane, cytoplasm, nuclear membrane and nucleoplasm of oocyte during the development and maturation of ovary, and spermatid and spermatozoa showed immuno negative reaction in testis. These results will provide a new mophological proof that LH and hCG may be directly involved in regulation of the gonadal development and maturation of protochordata. PMID- 12548776 TI - [Effect of PIC-BE on the expression of MDR-1 and bcl-2 genes in a multidrug resistance cell variant K562/ADM]. AB - The effect of PIC-BE on the expression of mdr-1, bcl-2 and bax genes and their protein products (P-gp, Bcl-2 and Bax) was observed respectively in a multidrug resistance (MDR) cell variant K562/ADM. The results showed that PIC-BE could significantly inhibit the expression of mdr-1 and bcl-2 genes at both mRNA and protein levels in K562/ADM cell line, and the effect was dose- and time-dependent within limited range. Under same condition, although PIC-BE could increase the expression of Bax slightly, there was no statistically significant difference. These results suggest that the reversal of the MDR of K562/ADM cell line by PIC BE may result from its effect on the expression of mdr-1, bcl-2 genes and their protein products. PMID- 12548777 TI - [Tissue specific differentiation of dorsal mesoderm in Xenopus mid-gastrula embryos]. AB - Genes involved in differentiation of notochord or muscle are expressed in specific regions of the involuted dorsal mesoderm in mid-gastrula Xenopus embryo. The presumptive notochord or the presomitic mesoderm have been cultured either in isolation or recombination to investigate whether these tissues have been determined. Cell differentiation was checked by specific markers of notochord (Shh) or muscle cell (desmin, myosin). The results show that the presumptive notochord can differentiate into vacuolated notochord with a weak expression of Shh, while the presomitic mesoderm differentiate into muscle cells with a normal expression of desmin and myosin in vitro. The same result was obtained when the two tissues have been cocultured. These data suggest that the cell fate of the involuted dorsal mesoderm in mid-gastrula has been determined, cells can differentiate according to their fates without further signals from the adjacent tissues, but no functional structures can be formed by these tissues in vitro. PMID- 12548778 TI - [Regulation of cell cycle by retinoic acid in gastric cancer cells]. AB - Retinoic acid can induce growth inhibition and apoptosis, and regulate cell cycle in many types of cancer cell lines. In this study, we investigated the role of all-trans retinoic acid (ATRA) and its mechanism of action in human gastric cancer cell lines. Our results demonstrated that ATRA effectively inhibited growth in three of four gastric cancer cell lines by induction of G0/G1 arrest, and did not induce apoptosis in four gastric cancer cell lines. In RA-sensitive cell lines, ATRA-induced G0/G1 arrest is associated with down regulaton of c-myc and hyperphosphorylated Rb expression, and up regulation of p21WAF1/CIP1 and p53 expression. There were no significant changes in cyclin D1 or CDK4 expression induced by ATRA. Futhermore, expression of these genes were not regulated by ATRA in ATRA-resistant gastric cancer cell line. These results indicate that growth inhibition, rather than apoptosis, is correlated with G0/G1 arrest of these cell lines, more important molecules related cell cycle, including c-myc, p21WAF1/CIP1, p53 and Rb, are involveed in regulation of cell cycle in gastric cancer cells. PMID- 12548779 TI - [Phenotypic analysis of TCR alpha beta+ CD4+CD8- medullary thymocytes]. AB - Phenotypic analysis of the medullary-type CD4+CD8- (CD4SP) thymocytes have revealed phenotypic heterogeneity within these cells. The phenotype of mature peripheral T cells is Qa-2+ HSA- CD69-, whereas in the medullary-type CD4SP thymocytes, the expression pattern of many markers were quite different, suggesting that the medullary-type CD4SP thymocytes may undergo phenotypic maturation. According to the results of two-color cytometry, seven discrete phenotypes were defined by the relative expression of Qa-2, HSA, CD69, 3G11 and 6C10: 3G11-6C10+CD69+HSAhi-->3G11+6C10+CD69+ HSAhi-->3G11+6C10-CD69+HSAint- >3G11+6C10- CD69-HSAint Qa-2(-)-->3G11+HSAlo/-Qa-2lo, at the same time, 3G11+6C10 CD69-HSAint Qa-2(-)-->3G11-HSAlo Qa-2(-)-->3G11-HSAlo/- Qa-2hi, the last two Qa-2 positive subsets could exit the thymus and home into periphery. PMID- 12548780 TI - [Activation of G protein on the membrane of TCS-sensitive cells]. AB - Trichosanthin (TCS), a Type I Ribosome Inactivating Protein isolated from the root tuber of Trichosanthes Kirilowii M. has various biological activities including abortion induction, anti-tumor and anti-HIV. The mechanism of TCS specifically killing sensitive cells has not been studied clearly. In this study, we initially found that there exists TCS-affinity molecule on Syncytiotrophoblast cells and Jar cells. Furthermore, by [35S]GTP gamma S Binding Assay, we find that TCS can activate G protein on the membrane of TCS-sensitive cells. These results indicate that on the membrane of TCS-sensitive cells exists TCS-specific receptor. PMID- 12548781 TI - [The antagonistic effect of naloxone on the epidermal conductivity of amphibian embryos]. AB - The immunocytological work revealed that in amphibia beta-endorphin is distributed in embryonic epidermis during development with prominent expression at late tail bud stage which coincides with the stage of epidermal conductivity of full strength. To see whether any relationship exists between the presence of beta-endorphin and the epidermal conductivity of amphibian embryos, naloxone of 1 and 2 mumol/L was used. It has been demonstrated that after 45-60 minutes in the naloxone solution the embryonic epidermal conductivity disappeared and 90-120 minutes after the beginning of naloxone treatment the epidermal conductivity was again recovered. It was suggested that the opiate peptides, such as beta endorphin, play some role in the epidermal conductivity of amphibian embryos. PMID- 12548782 TI - [Immunofluorescence studies on the localization of integrin-like protein in plant pollen tubes]. AB - The Strong fluorescence signals were obtained in pollen tube of Lilium davidii Duch with labeled anti-VnR integrin serum, and anti-beta 3, alpha v integrin subunit cytoplasmic domain serum separately. The highest density of immunolabel was in the tip of pollen tube. There was little or no immunolabel in control experiment using non-immune serum, second antibody alone and anti-FnR, LnR integrin serum separately. In pollen of Prunus persica f. rubro-piena Schneid, fluoresence signals were also obtained in tube using labeled anti-beta 1, beta 3 integrin subunit cytoplasmic domain serum separately and in apertures using anti beta 1 serum. Preliminary results show that during the germination of pollen and the growth of pollen tube, there may be integrin-like proteins in pollen tube, consisting of alpha v and beta s integria subunits in the pollen tube of Lilium davidii Duch, which is the receptor of vitronectin-like protein. PMID- 12548783 TI - [Regulation of fungal elicitors on the cell growth and biosynthesis of saponin of Panax ginseng cell suspension culture]. AB - Adding fungal elicitors to the Panax ginseng cell suspension cultures, the biosynthesis of saponin was obviously induced, the total productivity of saponin in cultures could increase more than 30% of the control. During elicitation, the accumulation patterns of saponin in suspension cultured cells were changed, the culture time for maximum biosynthesis of saponin was shortened 2-4 days comparing with that of the control, and about 80% of biosynthetic saponin in elicited cells was secreted into medium, meanwhile the uptake for sucrose in medium of cells was enhanced, and the disturbing of pH in medium was observed, which predicated that an ion exchange occurred between elicited cells and medium. PMID- 12548784 TI - [Direct and indirect somatic embryogenesis in Freesia refracta]. AB - Somatic embryogenesis can be induced in tissue cultures of Freesia refracta either directly from the epidermal cells of explant, or indirectly via intervening callus. In direct pathway, somatic embryos were in contact with maternal tissue in a suspensor-like structure. In indirect pathway, the explants first proliferacted to give rise to calluses before embryoids were induced. The two sorts of calluses were defined to embryogenic callus and non-embryogenic callus according to producing of somatic embryos. An indirect somatic embryo is developed from a pre-embryogenically determined cell. This kind of somatic embryo has no suspensor structure instead of a complex with maternal tissue. Somatic embryos have their own vascular tissues, and can develop new plantlets independently. PMID- 12548785 TI - [Localization with BrUTP labeling technique of RNA polymerase II transcription in meristematic cells of Allium cepa]. AB - The technique of pulse-labeling of cells with 5-bromouridine-5'-triphosphate (BrUTP) and immunological detection of the incorporated BrUTP is a newly developed experimental approach to localize nascent RNA polymerase II transcripts in the nucleus. This technique has been widely used since it was first reported in 1993, but most of the studies were carried out with fluorescence microscopy and animal cells, we are still limited in the knowledge about ultrastructural localization of RP II transcripts in the nucleus of eukaryotes, especially in that of plant cells. Attempts have been made in this paper to localize nascent RP II transcripts in the nucleus of meristematic cells of Allium cepa with the technique of pulse-labeling of BrUTP and immunoelectron microscopy. After labeling with BrUTP and detection with anti-BrdU antibody, a large number of gold particles representing RP II transcripts were observed in the periphery of chromatin domains, the gold particles in the periphery accounted for 92.70% of the total gold particles in the extranucleolar regions, and the gold density of the periphery was 75.42/micron 2, much higher than that of the center of the chromatin domains and the interchromatin domains which were only 5.89/micron 2 and 2.00/micron 2 respectively, indicating that RP II transcripts were being actively synthesized in the periphery of the chromatin domains. When the specimens were treated with alpha-amanitin which can specifically inhibits the activity of RP II, gold particles in the extranucleolar regions of the specimens were decreased dramatically and the gold density of the regions dropped strikingly from 44.60/micron 2 to 2.67/micron 2, confirming that the gold particles in the regions represent RP II transcripts. The distribution of nascent RP II transcripts in the nucleus of the intact plant cells is discussed. PMID- 12548786 TI - [Induction of programmed cell death by menadione in suspension culture of carrot cells]. AB - Menadione (VK3), a quinone that undergoes redox cycles leading to the formation of superoxide radicals, was found to induce cell death in suspension culture of carrot cells. The effect of menadione was in a dose-dependent manner. 100-800 mumol/L menadione caused 10-33 percent cell death. When concentration of menadione reached 1 mmol/L, 100 percent of cell death was observed. DNA cleavage, a hallmark of apoptosis was further studied. DNA ladders were observed in cells treated with 600 and 800 mumol/L menadione but not with lower concentration treatments where only very low percentage of cell death was found. There was no DNA ladders in the cells treated with 1 mmol/L menadion indicating that necrosis may occur. In situ detection of nuclear DNA fragmentation by TUNEL reaction revealed fragmented nuclear DNA in cells treated with 100-800 mumol/L menadion but not in cells treated with 1 mmol/L menadione. PMID- 12548787 TI - [A quick method to estimate the T-DNA copy number in transgenic rice using inverse PCR (IPCR)]. AB - In order to obtain single T-DNA copy transgenic rice, we have established a quick method to estimate the T-DNA copy number in transgenic rice using inverse PCR (IPCR). IPCR was used to amplify junction fragments, i.e. plant genomic DNA sequences flanking the known T-DNA sequences, which will help to estimate the T DNA copy number in transgenic rice. We have analyzed 20 transgenic plants of 15 transgenic lines. Most plants (12) contain one integrated T-DNA copy per genome, 3 plants contain two and 1 plant contains 3 copies. In 4 transgenic plants no T DNA copies could be detected using this method. The IPCR results were further tested by Southern analysis and sequence analysis. PMID- 12548788 TI - Effects of 16O+6 ion irradiation on human sperm spontaneous chemiluminescence, motility, acrosome reaction and viability in vitro. AB - Effects of 16O+6 ion irradiation with different doses on human sperm spontaneous chemiluminescence (SCL), motility, acrosome reaction (AR) and viability were examined. Spermatozoa were irradiated with 0, 0.25, 0.5, 1, 2, 4, 8, 16, 32, or 64 Gy 16O+6 ion beam at the energy of 3.17 MeV/u. After irradiation, samples were analyzed by SCL measurement at 1, 2 and 3 h of incubation; motility was determined by the transmembrane migration method within 2 h of incubation; the percentage of AR and viability was evaluated by the triple-stain technique at 3.5 h of incubation. The results showed: sperm SCL was significantly increased with irradiation doses and the lowest effective dose was 0.5 Gy; compared with controls, the transmembrane migration ratio of spermatozoa progressively elevated with irradiation doses at 0.5, 1, and 2 Gy; the percentage of sperm AR markedly increased in 0.5-4 Gy irradiation and the optimal dose was 2 Gy, and then significant decreased with further increase of irradiation doses; the viability had no significant change within 0.25-8 Gy, but was progressively decreased at 16, 32 and 64 Gy. These data suggested that heavy ion at low doses increased motility and AR, whereas had deleterious effects at higher doses, which are associated with free radical reactions induced by heavy ion irradiation. PMID- 12548789 TI - [Immunohistochemical study of the expression of fucosylated glycoconjugates in human lung carcinoma]. AB - Lewis X, Sialyl Lewis X and Sialyl Dimeric Lewis X are three fucosylated glycoconjugates on cell surface. With immunohistochemical method, the expression of the three structures in the original lung cancer tissues (with or without metastasis), adjacent tissues and metastatic lesions of lung carcinoma were studied. It was found that the three antigens were expressed with different intensity on the cell surface and in the cytoplasm of the lung carcinoma cell. However, there was no or only trace expression of these antigens in the adjacent tissues of lung carcinoma and normal lung tissues. Moreover, the original lesions of lung carcinoma with metastasis and/or poor differentiation expressed higher level of the three antigens than those without metastasis and/or with well or medium differentiation. Sialyl Lewis X was considered to be more closely related to the metastatic ability and differentiation of the lung carcinoma cell than the other two antigens, Lewis X and Sialyl Dimeric Lewis X. Furthermore, in the lymph nodes with lung carcinoma cell metastasis, there were expression of the three antigens with different degree, while in those lymph nodes without lung carcinoma cell metastasis, there was no expression of the three antigens. PMID- 12548790 TI - [Effect of alpha-fetoprotein on the growth of human hepatoma cells in vitro]. AB - The stimulatory activity of human alpha-fetoprotein (AFP) on the growth of mouse hepatoma-22 cells had been reported in our previous paper. The present work aimed at further investigation of the effect of AFP on human hepatoma cell growth by MTT colorimetric assay. The results showed that AFP could stimulate the growth of SMMC-7721 human hepatoma cells in vitro. The present results also showed that the stimulatory effect of AFP on the growth of SMMC-7721 cells was decreased by the anti-serum of AFP. The anti-AFP antibody alone could suppress the growth of SMMC 7721 cells. On the other hand, AFP and anti-AFP antibody had no effect on the growth of HL-60 human leukemia cells, indicating that the tumor cell growth stimulating effect of AFP was not simply due to non-specific addition of exogenous protein and this effect of AFP showed strict tumor cell specificity. In addition, MCF-7 human breast cancer cell growth was also promoted by AFP and inhibited by anti-AFP antibody. Because AFP cell-surface receptors have been detected in MCF-7 breast cancer cells, and AFP could also be produced and secreted by MCF-7 cells, the possibility may be considered: AFP may bind with its receptors on tumor cell membrane for the purposes of growth stimulation. PMID- 12548791 TI - [p53 gene expression of human hepatoma cell lines and their sensitivities to parvovirus H-1]. AB - DNA structure and expression of p53 gene in human hepatoma cell lines SMMC-7721, YY-8103 and a spontaneously transformed liver cell line L-02 were analysed using the following method: analysis of allelic losses on chromosome 17p, PCR/SSCP, Northern blot and immunoprecipitation. There was no point mutation found in the exons 4-9 of the p53 gene, and a low level of expression of p53 gene was detected in the three cell lines. These observations were in agreement to the reported results of the relevant experiment using the human hepatoma cell line QGY-7703. Sensitivities of these cell lines and other eight human hepatoma cell lines (QGY 7703, PLC/PRF/5, Tong/HCC, Huh-7, FOCUS, Hep3B, SK-Hep-1, HepG2) with known p53 backgrounds to parvovirus H-1 was assayed using MTT method. Abnormality in the structure and/or function was observed in all of the cell lines examined except HepG2. The cell line HepG2 with normal structure and function of the p53 gene was found to be the least sensitive to H-1 in comparison to all the cell lines which have defeated structure and/or function of the p53 gene. The present study serves as a preliminary evidence that enhancement of the sensitivity of human hepatoma cell lines to H-1 is correlated to the abnormality of the structure and/or function of the p53 gene. PMID- 12548792 TI - [Biosynthesis of a single peptide chain containing human chorionic gonadotropin beta and C3D of complement]. AB - In view of the strong immunity-enhancing function of HEL-C3d3 designed by Dr. Paul W. Dempsey, we made our efforts to produce a similar recombinant protein of hCG beta. With polymerase chain reaction, we introduced a Bam HI restriction site into the 3' terminal of hCG beta cDNA. The new cDNA and its terminal's correctness has been confirmed by sequencing. Then we have it covalently attached to the C3d3 cDNA at the pre-designed Bam HI/Bgl II site. Having the chimeric DNA correctly cloned into the protein nuclear polyhedrosis virus (AcNPV) expression vector pVL1393, we constructed the expression vector pVL1393-(hCG beta-C3d3). The insect cells were co-transfected with the expression vector and linearized nuclear polyhedrosis virus DNA, and recombinant viruses AcNPV-(hCG beta-C3d3) were screened out. Through anti-hCG beta immunoaffnity chromatography, the recombinant hCG beta-C3d3 chimera polypeptide was purified from culture supernatant of insect cells infected by the recombinant viruses. In RIA test, the expressed product competitively inhibits the binding of 125I-hCG beta to hCG beta antibody. On SDS-PAGE and Western blot, the recombinant peptide hCG beta-C3d3 obviously appears to be with a molecular weight of 116KD. Therefore, we arrive at a conclusion that it has a normal immunogenic ability. PMID- 12548793 TI - [Comparative study of dependence of the cell proliferation of Saccharomyces cerevisiae and Schizosaccharomyces pombe on Ca2+]. AB - Under the same experimental conditions, exogenous Ca2+ had no effect on the proliferation of S. cerevisiae, but it could obviously stimulate the proliferation of S. pombe. Ca2+ chelator EGTA had no inhibition effect on the proliferation of S. cerevisiae, but it apparently inhibited the proliferation of S. pombe and the inhibition could be effectively overcome by adding Ca2+. Non special ion chelator EDTA could inhibit the proliferation of both S. cerevisiae and S. pombe, but the inhibition could not be overcome by adding Ca2+. The results above directly showed that the dependence of the proliferation of the two kinds of yeast on exogeneous Ca2+ was different. The growth rate of S. cerevisiae was about 3 times that of S. pombe and the proliferation of S. cerevisiae was independent on the exogenous Ca2+, which was similar to transformed cells. Therefore, in order to understand the relationship between the disorder of cell cycle and cell transformation, it was very important to study the mechanism of different effects of exogenous Ca2+ on the proliferation of the two kinds of yeast. PMID- 12548794 TI - [Establishment of studying system cloned vernalization-putative cDNA clone by means of differential display PCR]. AB - Differential display PCR has been developed as a tool detecting and characterizing specially expressed gene in eukaryotic cells recently. In this paper, a studying system was established which applied the novel approach of differential display PCR to clone vernalization-related cDNA clone. Here methodological details were provided which included the purification of total RNA, removal of contaminated DNA, reverse transcription of cDNAs, parameters of PCR, electrophoresis of amplified cDNAs, recovery and reamplification of cDNAs. It was also identified a vernalization-putative cDNA clone (VPC) of VPC28 which was only expressed at the key stage of vernalization for 20 d in winter wheat. These results moreover made it readily applicable to a broad spectrum of similar studies. PMID- 12548795 TI - [The role of calcium in IAA-induced swelling of protoplasts isolated from hypocotyl of etiolated mung bean seedlings]. AB - This paper studied on the role of calcium in IAA-induced swelling of protoplasts isolated from hypocotyl in etiolated mung bean (Phaseolus radiatus L.) seedlings. Protoplasts incubated in CaCl2-bearing medium without hormone maintained a constant volume and a consistent intensity of 45Ca2+ radioactivity. To treat with IAA, they began to swell and continually swelled to the maximum volume 30 minutes later (Fig. 2). However, the protoplasts could not swell when IAA was added into the medium without CaCl2 (Fig. 1). It was suggested that Ca2+ may be necessary for IAA to induce protoplast swelling. And also, IAA enabled the protoplasts to swell in less extent with K+, Zn2+, Ba2+ or Mg2+ instead of Ca2+ (Fig. 3). Radioisotope experiments showed that K+ influx increased when K+ replaced Ca2+ (Fig. 4), and water absorption plays a role in the swelling (Fig. 5). 45Ca2+ accumulation in protoplasts treated by IAA was much higher than that of control, and the time course of 45Ca2+ accumulation was similar to that of protoplasts swelling (Fig. 6). 45Ca2+ level and the swelling of protoplasts sharply declined when EGTA, verapamil or LaCl3 was added into the medium (Table 1, 2 and 3). These results indicated that Ca2+ may play an important role in IAA-induced swelling. PMID- 12548796 TI - [Vascular bundle specific expression of iaaL gene affects the generation frequencies of transgenic tobacco]. AB - A vascular bundles specific expressing vector pBAL1 with a promoter AQ630 from rice phenylalanine ammonialyase gene and a gene encoding indoleacetic-lysine synthytase from Pseudomonas syringae subsp. savastanoi was constructed. Affirmed by Southern blotting and RTPCR analysis, the AQ630-iaaL transgenic plants show increasing shoots-regeneration frequency of young stem explants on hormone-free 1/2 MS medium and lower sensibility to IAA when roots were induced from the root explants on the media containing different concentrations of IAA compared to untransformed plants. PMID- 12548797 TI - [Enhanced resistance of transgenic tobacco expressing Shiva A gene against bacterial wilt disease (Pseudomonas solanacearum pv tabaci)]. AB - Shiva A gene was introduced into tobacco mediated by Agrobacterium tumefaciens. Transgenic plants show enhanced resistance against bacterial wilt disease (Pseudomonas solanacearum pv tabaci). Compared with control plants, the disease indexes of transgenic tobacco plants Sc-2 and Sc-6 drop about 42.1% and 60.6%. PMID- 12548798 TI - [HarpinPss elicited early defense responses of tobacco cells and involvement of calcium]. AB - HarpinPss can induce hypersensitive reaction (HR) in tobacco leaves. As superoxide dismutase can inhibit but catalase can not inhibit the development of HR induced by harpinPss, superoxide anion is required for this response. HarpinPss can also induce the release of active oxygen and extracellular alkalinization, two early defence responses in tobacco suspension culture. Diphenylene iodoium, can completely inhibit the induction of HR in tobacco leaves, and the release of active oxygen in the suspension culture system, superoxide anion in these system may be produced by the activation of NADPH oxidase. Ethyleneglycol-bis (beta-aminoethyl) N, N, N'N'-tetraacetic acid (EGTA) can inhibit the development of harpinPss-induced HR and these two early defence responses in suspension culture system. Adding Ca2+ into the medium again, these responses can return to normal level in a short time. Lanthanum chloride, verapamil, neomycin, U-73122, and LiCl can also inhibit these harpinPss-induced responses. Therefore, the influx of Ca2+ mediated by calcium channel and the release of Ca2+ from internal Ca2+ pool may be involved in the two early defense responses induced by harpinPss. Cycloheximide and actinomycin D have no effect on the release of active oxygen but can inhibit harpinPss-induced HR even added them in the intermediate process for inducing HR. It indicates superoxide is just a trigger for HR, and HR is a more complex process that needs the sustained expression of some genes. PMID- 12548799 TI - [Involvement of anion channel in signal transduction of early defense responses elicited by harpinPss in tobacco]. AB - No matter when anion channel inhibitors, DIDS (4, 4'-diisothiocyanatostilbene-2, 2'-disulfonic acid) and A9C (anthracene-9-carboxylic acid) added (before, at the same time of or after harpinPss treatment), they can inhibit harpinPss-induced hypersensitive response in tobacco seedlings and release of active oxygen and extracellular alkalinization in tobacco suspension cells. DIDS and A9C also inhibit harpinPss-induced Ca2+ influx. In all these cases, DIDS is more efficient than A9C. It is postulated that anion channel positively regulates calcium channel in plasma membrane, and harpinPss may function through signal transduction mediated by anion channel and calcium channel to regulate cellular Ca2+ concentration and defense responses. PMID- 12548800 TI - [Preliminary studies on tissue culture and agrobacterium-medicated transformation of Brassica campestris ssp. chinensis]. AB - The hypocotyls and cotyledons of the asepetic seedling of Brassica campestris ssp. chinensis L cv. Pudongaijiecai) were used as explants for tissue culture. Adventitious buds were differentiated on modified MS medium supplemented with TDZ 1-2 mg/L, NAA 0.2-1 mg/L and AgNO3 7.5 mg/L. The percentage of explants which formed buds of cotyledons was about 56%, and that of hypocotyls was about 37%. When the regenerated explants were transferred onto MS medium with 2 i.p. 5 mg/L and NAA 0.1 mg/L for two weeks, whole plantlets were obtained by culturing the regenerated shoots on 1/2 MS medium with NAA 0.1 mg/L. Agrobacterium tumefaciens strain (LBA 4404/PBI 121) carrying the GUS gene and Npt II gene was used for transformation. After 2 days of coculture, the hypocotyls and cotyledons were transferred onto regenerated medium containing CP 300 mg/L for bud formation. After 4-5 weeks, the differentiated buds were transferred onto selection medium with CP 200 mg/L and Km 10 mg/L for 1 month, then the green shoots were transferred onto the rooting medium containing Cef 100 mg/L and Km 20 mg/L. 4-5 weeks later, plantlets with Km resistance were obtained and some of them showed higher enzymatic activities of beta-glucuronidase than control ones. PMID- 12548801 TI - [In commemoration of professor Luo Shi Wei]. PMID- 12548802 TI - [Effect of L1ECD on mouse primarily cultured neurons and construction of transgenic mice specifically expressing L1ECD in brain]. AB - Neural cell adhesion molecule L1 is an important molecule mediating cell-cell interactions during the development of nervous system. L1 can promote axonal outgrowth and is related with nerve cell migration, and therefore L1 plays an important role both in the development and maintaince of the nervous system. In humans, mutations in the L1 gene can lead to mental retardation, spastic paraplegia, hydrocephalus, and other developmental abnormalities. The molecular mechanisms of mutations in L1 gene to induce inherited neurological diseases are not clear. In present investigation, a transgenic DNA of mouse L1 extracellular domain (L1ECD) was constructed by adding a stop codon to the end of L1ECD cDNA and then putting it under the control of CAMK II promoter, which is active specifically in the brain. To verify this construct, L1ECD cDNA was subcloned into an expression vector pCEP4 and then transfected the C6 cells. The expression of L1ECD cDNA in C6 cells was confirmed by Northern blotting and the effects of L1ECD on the growth rate and morphology of C6 cells in vitro as well as primarily cultured neurons were observed. The L1ECD constructs were microinjected into the fertilized zygotes of C57BL/6 mice. The transgenic mice thus produced were identified by Southern and Northern hybridization analysis. The results demonstrated that the L1ECD was integrated in the genome of transgenic mice and expressed specifically in the brain. PMID- 12548803 TI - [Study on the interaction between the 5' proximal region of mGAT-1 and nuclear proteins by the method of SPR]. AB - The DNA fragment (named F182) corresponding the position of -1775(-)-1594 in the mouse GABA transporter 1 (mGAT-1) 5' proximal region was amplified by PCR. Then the DNA was immobilized to the surface of sensor chip SA5 via biotin-streptavidin linkage. The interaction between the F182 on SA5 and nuclear proteins from mouse liver and kidney was studied by the method of SPR with Biosensor of BIAcore-1000 respectively. The Binding between F182 and two nuclear proteins was definitely and specifically and both with the apparent dissociation rate of about 1.4E-5/s. Competitive experiment revealed that a conserved sequence within F182 had the main contribution to the binding event. PMID- 12548804 TI - [The development of mouse bioreactor expressing human tissue plasminogen activator (tPA) in mammary gland by transfecting spermatozoa in testicular duct]. AB - The most established methods for development of transgenic animals are the microinjection of DNA into the fertilized eggs, but it is still a procedure of certain complexity and high cost. Therefore, the idea of using sperm as a vehicle to carry exogenous DNA into eggs is very attractive, and there have been some successful reports. Though the methods are rather simple they sometimes have low reproducibility. To improve the technique we transinfected the spermatozoa in testicular duct, not in vitro, to produce mice which expressed human tissue plasminogen activator (tPA) in mammary gland. The results demonstrated that: (1) 5 transinfected mice mated 10 female mice in 10 days after operation, (2) 79 founders were developed and 42 survived, (3) using PCR to detect foreign DNA integrated into the genome of founders, 7 out of 42 founders were positive (16.67%), (4) The expression level of tPA was 48-80 ng/ml in the milk of 5 PCR positive founders and (5) the foreign DNA integrated into the genome was detected in 2 out of 4 1st offspring by PCR technique. PMID- 12548805 TI - [Isolation and cloning of human beta 1, 4-galactosyltransferase III gene]. AB - Using the conservative nucleotide sequences encoding the catalytic domain of the beta-1, 4-GalT genes in human, bovine, mouse, chick and snail as probes to search the NCBI GenBank EST database, several ESTs with high homology were obtained. Primers were designed in the flanking sequence of EST contig. Using the PCR product amplified in human placenta cDNA library as probe to perform "walking" hybridization with human placenta cDNA library, a cDNA fragment with the length of 1,907 bp was cloned. It contained an open reading frame (ORF) with the length of 1,179 bp, which encodes 393 amino acid residues. The deduced amino acid sequence of this gene shares 43.8% identity to the human beta-1, 4-GalTI and 60.9% in the catalytic domain especially. The expression mapping showed that it was expressed in human most tissues with a single 2.4 kb transcript, but the relative expression level of the transcript are vary. While this gene was mapped on chromosome 1 using the cDNA hybridization with human/rodent hybrid cell line DNA Southern blot panel. PMID- 12548806 TI - [Analysis of the binding of GATA factors to the regulatory elements of human beta globin gene in hydroxyurea-induced and uninduced HEL cells]. AB - HEL cells, a human erythroleukemia cell line, expressing mainly the gamma-globin genes, small amount of epsilon-globin gene, but not beta-globin gene. Our previous studies demonstrated that beta-globin gene could be expressed in HEL cells induced by hydroxyurea. However, the molecular mechenism is still unknown. Here the binding patterns of GATA factors (GATA-1 and GATA-2) to the regulatory elements of human beta-globin gene were examined with the nuclear extracts from hydroxyurea-induced and uninduced HEL cells. Our results showed in EMSA assay that GATA factors could bind to the core sequence of HS2(-10681 to -10971 bp), the 3' flanking sequence of HS2 core(-10323 to -10680 bp) and the promoter of human beta-globin gene(+20 to -112 bp). However, the binding patterns between hydroxyurea-induced and uninduced HEL cells were different. Furthermore, by using Western-blot analysis, our data showed that the amount of GATA-2 was decreased in hydroxyurea-induced HEL cells. In contrast to GATA-2, the amount of GATA-1 was increased in hydroxyurea-induced HEL cells. These results showed that the different members of GATA family might play different roles during the differentiation of erythrocytes. GATA-1 may stimulate the differentiation of HEL cells, while GATA-2 can probably inhibit the differentiation of HEL cells. PMID- 12548807 TI - [Establishment and differentiation of murine EG cell lines derived from primordial germ cells]. AB - Primordial germ cells (PGC) were isolated from 8.5, 10.5, 12.5 days post coitum (dpc) embryos of F1 (Balb/c x ICR), C57BL/6J, 129/svJ, 129/sv-ter mice, and cultured on mitotically inactive MEF or STO feeder layer cells with addition of leukemia inhibitory factor, stem cell factor and basic fibroblast growth factor in cultures. PGCs formed densely packed and AKP positive colonies with pluripotential marker gene (oct-4) expression resembling undifferentiated ES cells in morphology and growth pattern. Five EG cell lines derived from PGCs were established: EG1(8.5 dpc, F1), EG2 and EG3 (8.5 dpc, C57BL/6J), EG4 (10.5 dpc, 129/svJ), EG5 (10.5 dpc, 129/sv-ter). No long term culture was obtained from 12.5 dpc PGCs of 29 embryos. All five EG cell lines cultured on feeder layer cells or in LIF containing medium still remain undifferentiated state at 15 th passage. Under appropriate conditions, EG cells formed embryoid bodies in suspension culture and multiple types of differentiated cells in monolayer culture. When these EG cells were injected in nude mice, they formed teratocacinomas containing differentiated cells such as cartilage, neural tissue and epithelium. These results show that EG1-5 cell lines derived from 8.5, 10.5 dpc embryos are pluripotential. PMID- 12548808 TI - [RAPD method for the identification of intergeneric asymmetric somatic hybrid plants of wheat]. AB - Randomly amplified polymorphic DNA (RAPD) method was used to identify the hybrid nature of three kinds of intergeneric asymmetric somatic hybrid plants of wheat: wheat (Triticum aestivum) + Haynaldia villosa, Wheat + Leymus chinensis and wheat + Agropyron elongatum. It was shown from the electrophoresis profiles that the genome of somatic hybrid plants contained specific section genome of both parents after DNA amplification with arbitrary primers. A specific RAPD product (DNA fragment of 0.77 kbp) of A. elongatum generated with primer OPJ-12 was isolated, purified, labeled and used as a probe. Southern blot from OPJ-12 primer-generated specific section genome of the hybrid (T. aestivum + A. elongatum) hybridized to this probe (0.77 kbp) proved that they are homologous in nature. This paper also discussed the advantage of RAPD method in identification of hybrid plants, especially asymmetric somatic hybrids. PMID- 12548809 TI - [Studies on the application of antifreeze proteins in cryopreservation of rice suspension cells]. AB - AFP from winter flounder was utilized in cryopreservation of plant cells. During cryopreservation of rice suspension cells by two-step method, AFP at 0.01 mg/ml damaged the cells extremely. The data obtained at relatively high concentration, however, decreased the variability of survival rate. During vitrification of rice cells, AFP at 0.2 mg/ml enhanced the viability. However, high concentration AFP (> 5 mg/ml) decreased the recovery rate. Studies indicated that the results of application of AFP in cryopreservation were closely related to the concentration of cryoprotectant. The amount of ice crystal in environment, the concentration of AFP and cryoprotectant, and the composition of plasma membrane were several key factors affecting the results of AFP application. In mechanism analysis, the authors suggested that on one hand AFP can interact with ice crystal, which inhibits ice recrystallization and prevent the cells from devitrification. On the other hand, AFP also can interact with cell membrane, resulting in the ice growth around the plasma membrane. PMID- 12548810 TI - [Localization of SC35 in the nucleolus and nucleoplasm of the root meristematic cells of Vicia faba]. AB - SC35 is a non-snRNP spliceosome component purified from mammalian cells by Fu and Maniatis in 1990. In vitro splicing assays showed that SC35 plays a key role in splicing site selection and ATP-dependent pre-spliceosome assembly. In the mammalian nucleus, SC35 has been localized to distinct and dynamic nuclear domains: immunofluorescence observations revealed the presence of SC35 in speckles distributed in various regions throughout the nucleoplasm, which, as identified with immunoelectron microscopy, correspond to the interchromatin granules (IGs) and perichromatin fibrils (PFs). However, there has been no report regarding the presence and distribution pattern of SC35 in higher plant nuclei. Engage in such studies will surely contribute to our understanding of RNA processing and the spatial organization or structure basis of this process in higher plant. In this article, we studied the distribution pattern of SC35 in the nucleus of the root meristematic cells of Vicia faba by immunoelectron microscopy. After immunolabeling with anti-SC35 mAb and protein A-colloidal gold, IGs and PFs in the nucleoplasm and dense fibrillar component (DFC) of the nucleolus were heavily labeled with gold particles, while only a few of the gold particles were found in fibrillar centers (FC) and nucleolar vacuoles (NV) of the nucleolus and the central domains of the condensed chromatin. Densities of gold particles in the areas of DFC and the area of IGs plus PFs were 65.89/microns 2 and 36.28/microns 2 respectively, much higher than that of the central domain of condensed chromatin and that of FC plus NV, which were only 5.90/microns 2 and 6.26/microns 2 respectively. This indicates that DFC of the nucleolus and the area of IGs plus PFs of the nucleoplasm are enriched with SC35 or SC35-like protein. The distribution pattern of SC35 or SC35-like protein in the nucleoplasm of Vicia faba is similar to that of the mammalian nuclei. To the authors' knowledge, it is a new finding that SC35 or SC35-like protein exists in the nucleolus. PMID- 12548811 TI - [Induction of embryogenic callus of loose skin mandarins]. AB - Young ovules from 3 cultivars and undeveloped ovules in mature fruits from 8 cultivars of loose skin mandarin of Citrus were cultured on 4 different media respectively to induce embryogenic calli. Results showed that the combination of EME(MT + 500 mg/L malt extract) and MKT (EME + 10 mg/L KT) media performed well in the induction of embryogenic calli from young ovules; MGS(EME + 1 mg/L GA3 + 40 mg/L sulfate adenine) medium was better than MDB (MT + 0.01 mg/L 2,4-D + 0.1 mg/L BA) medium in inducing calli from the undeveloped ovules, and the darkness was conducive to the induction of embryogenic calli. There was no chromosome number variation in the induced calli. All of the examined cells were diploid with 2n = 2x = 18 chromosomes. PMID- 12548812 TI - [Quantitative stereologic analysis of metabolize dynamic of protein and DNA during somatic embryogensis in Lycium barbarum L]. AB - The protein and DNA content in somatic embryos of Lycium barbarum L were analysed using quantitative stereologicai procedure formed from Cell Morphometry. The photos were transformed into digital images by the Sharp JX-610 scanner. Then the quantity of protein and DNA was count applying digital images processing software. By the result we discuss the relationship between changes in protein and DNA metabolize dynamic and somatic embryogensis. PMID- 12548813 TI - [The reactions in ginseng cells induced by elicitor derived from digesting ginseng cell walls by cellulase]. AB - After treatment of the elicitor derived from digesting ginseng cell walls by cellulase, the starch grains were degraded, and a large number of spherosomes were produced in Ginseng cells. In the mean time, the activity of amylase in the cells was increased and the activities of esterases in the cells were reduced; in addition, the content of soluble sugar and the content of lipoid were increased. These indicate that the emergency reactions and the adjustment of metabolism of lipoid were taken placed in the cells elicited. Besides, after the treatment with certain concentrations of elicitor, the content of ginseng saponin and the activity of phenylalanine ammonia lyase (PAL) were increased in ginseng cells. On the contrary, the activities of peroxidase (POD) and polyphenoloxidase (PPO) were reduced. PMID- 12548814 TI - [Microdissection of M chromosome in Vicia faba and its library construction]. AB - Microdissection and microcloning technique was employed to construct the library of M chromosome in Vicia faba. The M chromosomes were microdissected with a micromanipulator and were put into a 0.5 ml Eppendorf tube, then digested with Sau3A. Sau3A linker adaptors were ligated to the end of chromosome DNA fragments, and two rounds of PCR were carried out with one chain of linker adaptor as the primer. The PCR products ranged in size from 300 base pair (bp) to 3000 bp with predominant fragments from 500 bp to 1500 bp. Southern hybridization analysis confirmed that PCR products originated from Vicia faba genome. The second round PCR products were cloned and about 102,000 recombinants were obtained. 118 recombinants were selected randomly for analysis. The inserts ranged in size from 150 bp to 3000 bp with an average of 690 bp. Dot blot was carried out for 100 clones with DIG labeled Vicia faba genome DNA as probes. The result revealed that 51% were low and unique copy sequences, 49% were repetitive sequences. M chromosome DNA library has not been reported before. PMID- 12548815 TI - [The basic model and its peculiarity of relaxation theory in chromatography]. AB - According to the basic idea of relaxation theory in chemical dynamics and the moving character of the sample molecules in a chromatographic column, a new transfer model was established. It was supposed that solute molecules were in equilibrium between the stationary phase and mobile phase, and transferred in the mobile phase by a way of jumping. The generating function was used to investigate the non-continuous dynamic transfer differential equations, and the basic profile of solute's distribution in the column is gained [formula: see text] Based on this equation, the effect of transfer coefficient on the profile had been explained. The results show that relaxation time is an important factor which is related to the capacity factor and diffusion coefficient, and the number of jumping is a similar parameter to the plate number. The digital calculation method was used to study the influence of different physical parameters on the profile. The continuous form of the profile was also compared with the one from other methods. PMID- 12548816 TI - [Selectivity tuning in multi-binary eluents for reversed-phase liquid chromatography (RPLC)]. AB - In this article, the retention equation and the relationship between retention parameters and the parameters of molecular structure deduced from statistical thermodynamics in RPLC have been used to explain the difference of selectivity towards a particular species of compounds polycyclic aromatic hydrocarbons (PAHs). Methanol/water, acetonitrile/water and isopropanol/acetonitrile have been provided in advance, then the retention behaviors of sixteen PAHs under three binary solvent systems have been investigated. It is found that each pair of binary solvents of methanol/water, acetonitrile/water and isopropanol/acetonitrile has its own unique selectivity. The best selectivity obtained for acenaphthene and fluorene is methanol/water system for fluoranthene and pyrene is acetonitrile/water, and for benzo[g,h,i]perylene and dibenzo[a,h]anthracene is isopropanol/acetonitrile. So a three-stepwise gradient elution of multi-binary mobile phase can be chosen for separation of 16 PAHs. PMID- 12548817 TI - [Research of polymer coating in solid phase microextraction]. AB - The solid phase microextraction (SPME) is a new extraction technique which has been developed rapidly in 1990s. It is a fast, simple, solventless and sensitive method for analyzing environmental samples. However, the solid phase materials of SPME are relatively limited. This paper presents the research work on a new type of solid phase, polymethylvinylsiloxane (PMVS), used in SPME. The phase with 1% vinyl content could be used in light curing. The PMVS solution was coated on the surface of silica fiber and was quickly cured by UV. Two fibers with 87 microns and 44 microns thick PMVS coatings were prepared. They were compared with commercial polymer coating SPME fibers through Headspace SPME Gas Chromatograph (HS-SPME-GC). The adsorption and desorption kinetics of PMVS were studied. The HS SPME system was optimized and the differences between HS-SPME and HS were investigated as well. The results indicated that PMVS is efficient in extracting volatile and semi-volatile organic compounds and PMVS showed the high thermostability and easy-coating properties. The detection limit of SPME using 88 microns PMVS coating was about 1-5 micrograms/L. PMID- 12548818 TI - [The relationship between the thin-layer chromatographic retention values and the molecular structures of a group of amino acids by using back-propagation artificial neural networks]. AB - The relationship between the thin-layer chromatographic retention values and the molecular structures of fifteen amino acids was studied by using back-propagation artificial neural networks (ANNs). In this paper, firstly, lots of parameters of amino acids have been determined, accumulated and computed. Then, correlation coefficients of all parameters were computed by taking advantage of correlation analysis. Taking the correlation coefficient approaching one as the criteria in the correlation analysis, all parameters were classified into three kinds. Three parameters were selected from each kind, respectively, to consist of one group. Then, the optimized groups of parameters, which have clearer physicochemical meanings, were used as the inputting parameters of artificial neural networks. Correlation coefficients of experimental retardation values and those calculated by using ANNs were computed and showed good agreement. The present work shows that the ANNs method may take an important role in the study of the relationship between TLC behavior and compound structure. PMID- 12548819 TI - [The relationship between thin-layer chromatographic retention values and molecular structures of phenol and aniline derivatives]. AB - The Rf values of 24 phenol and aniline derivatives were determined on silica gel layer with three solvent mixtures of hexane-ethyl acetate in different proportions as the mobile phases. The compounds were divided into two species having different features by using the oblique factor analysis. It was found that in the first species there is no methyl group on their benzene rings and in the second all have methyl group in their benzene rings, which results in higher Rf values of the latter than those of the former. It was also found that the sequence of the groups on the benzene rings which make the Rf values of the compounds higher was --CH3 >--OH >--NH2 and the compounds having the ortho substituted groups give the higher Rf values than para- and meta-substituted ones due to the formation of hydrogen bond within their molecules. The multi-parameter regression analyses of Rf values with the topological indexes degree B and M for the two species showed rather good correlation between the TLC retention values and the molecular structure of the compounds. The findings are of great practical significance. PMID- 12548820 TI - [Temperature effect and temperature gradient technology in capillary electrophoresis]. AB - A review is given here to describe the temperature effect and temperature gradient technology in capillary electrophoresis (CE). The temperature effects on the analysis of DNA, protein peptides and amino acids, enantiomers, carbohydrates and other substances are discussed respectively. The theory of temperature effect is also mentioned. The importance of the temperature gradient in CE, the classifications of gradient technology in CE, the potential applications and different method development of the temperature gradient are described. Fifty-one references are cited. PMID- 12548821 TI - [Analysis of chiral compounds by micelle electrokinetic capillary chromatography]. AB - An overview on applications of micelle electrokinetic capillary chromatography (MECC) in chiral compounds analysis is presented. Ionic chiral surfactants such as bile salts, long-alkyl-chain surfactants and high-molecular-weight surfactants, which can migrate with electroosmosis and its electrophoretic mobility, and interact with the solutes, are employed as chiral selectors. Addition of electrically neutral chiral selectors such as cyclodextrins is also successful in MECC separation. Theory of MECC enantiomer separation is briefly described. PMID- 12548823 TI - [Study on thermodynamic interactions between polyoctene-1 and solvents using gas chromatography]. AB - Polymer of long chain alpha-olefins is a kind of comb-like polymer with special properties. In this paper, the thermodynamic interactions of polyoctene-1 with eight solvents have been studied by using gas chromatography. The eight Flory Huggins interaction parameters chi 1 were calculated. The results indicated that chi 1 decreases with increase of the carbon number of linear alkane, but when the carbon number is larger than 8, chi 1 tends to increase with the carbon number. It was also shown that the interaction of polyoctene-1 with cyclohexane is the strongest among all the studied solvents, and that with n-octane is the strongest among the linear alkanes. The relation of interaction parameter (polyoctene-1/n octane) with temperature T was also determined: chi 1 = 1.697-523.1/T. It was shown that the dissolution of polyoctene-1 in n-octane is an exothermic process with an enthalpy change of -4.84 kJ/mol. The solubility parameter delta 2 of polyoctene-1 is 27.42 (J/mL)1/2, determined from the interaction parameters chi 1. PMID- 12548822 TI - [Application of supercritical fluid extraction (SFE) on environmental analysis]. AB - In this paper, publications from 1993 are classified according to the type of matrix from which the environmental analytes are extracted. Although the most applications of SFE are used for the solid, some analysts have concerned with aqueous and gaseous media. Various aspects of investigation of environmental potential carcinogen, such as, polycyclic aromatic hydrocarbons, polychlorinated biphenyls, dioxins, pesticides, phenols and other hazardous and toxic pollutants in environmental samples have been reviewed with 55 references. The coupled techniques of SFE have also been introduced in this paper. PMID- 12548824 TI - [Gas chromatography-infrared spectroscopy(GC-IR) analysis of the oxidation products of cyclohexene]. AB - Gas chromatography-infrared spectroscopy technique integrates the excellent resolution ability of GC with the structural information of IR. Based on the GC and IR spectra, the complicated products of a reaction can be analyzed quantitatively and qualitatively. From the analysis of the products, the reaction process and its mechanism can be elucidated. In this work, analysis by GC-IR of the oxidation products of cyclohexene catalyzed by zeolite TS-1 in H2O2 was performed. The products are 2-cyclohexenone, 2-cyclohexen-1-ol and 2,3-epoxy-1 cyclohexanone. 2,3-Epoxy-1-cyclohexanone is an unexpected product. It must be a further oxidation product of 2-cyclohexenone. From the analysis, the reaction mechanisms were presumed. PMID- 12548825 TI - [Study on chemical constituents of essential oil of Michelia leaves by gas chromatography-mass spectrometry]. AB - The essential oil in Michelia leaves was extracted by steam distillation. The oil obtained was dried with anhydrous magnesium sulfate. The chemical constituents were analyzed by GC-MS. Thirty three peaks were separated by GC, and 27 of them were identified by MS with NBS mass spectral data and Wiley/NBS registry of mass spectral data. The structure of linalool was further verified by GC/FTIR. The identified constituents represent 97% of the peak area of the essential oil on FID. The major chemical constituent of them are linalool, trans-caryophyllene and diethyl-o-phthalate. PMID- 12548826 TI - [Enantiomeric resolution of trans-1,2-disubstituted cyclopropane by high performance liquid chromatography]. AB - The enantiomeric resolution of racemic mixtures of thirteen trans-1,2 disubstituted cyclopropane was achieved with HPLC by using Chiralcel OD and Chiralcel OJ as chiral stationary phase and hexane/2-propanol mixtures with different concentrations as eluent. The chromatographic parameters of these racemates on Chiralcel OD and Chiralcel OJ columns were examined. The results showed that enantiomeric resolution ability of the racemic mixtures of trans-1,2 disubstituted cyclopropane on Chiralcel OD and Chiralcel OJ was strongly dependent on the site and the property of substituents on the benzene cycle. On the other hand, some racemated of trans-1,2-disubstituted cyclopropane with aliphatic group were also separated on the columns. The results showed that the chiral recognition process was the sum of all the possible chiral interaction (the dipole-dipole, hydrogen bond-hydrogen bond and pi-pi interactions) between the chiral stationary phase and trans-1,2-disubstituted cyclopropane. This method has been applied to identify the optical purity of trans-1,2-disubstituted cyclopropane. PMID- 12548827 TI - [Purification of outer membrane proteins in Pseudomonas aeruginosa by high performance ion-exchange liquid chromatography]. AB - Pseudomonas aeruginosa PAO1 was used in this study. Isolation of outer membrane was accomplished by treating the cell envelope with EDTA and lysozyme, followed by centrifugation. The outer membrane (10 mg of protein) was mixed with 34 mmol/L octyl beta-glucoside-5 mmol/L EDTA-10 mmol/L Tris-HCl (pH 8.0) and subjected to supersonic oscillation for 2 min. The centrifuged supernatant (100 kgf for 30 min at 20 degrees C) was applied onto a DEAE ion-exchange high performance liquid chromatographic column (TSK gel-DEAE-5PW column, 0.75 cm x 7.5 cm i.d.) that was equilibrated with a solution of 10 mmol/L Tris-HCl buffer (pH 8.0) containing 2.5 mmol/L beta-C12E8 and 1 mmol/L EDTA. The column was washed with the same solution and eluted with a linear gradient of 0-0.5 mol/L NaCl in the same solution and fractions A, B, C were collected. Proteins in these fractions were analyzed by SDS-polyacrylamide gel electrophoresis and quantified by the method of Lowry et al. Protein E(Mr 43,000), G(Mr 25,000) and H (Mr 19,000) flowed through the column without adsorption in fraction A. Protein C(Mr 70,000), D(Mr 46,000) and a small amount of F (Mr 34,000) were eluted in fraction B. Fraction A was concentrated with ultrafiltration and applied again onto a DEAE ion-exchange HPLC column equilibrated with 10 mmol/L Tris-HCl buffer, pH 8.0, containing 34 mmol/L beta-C12E8 and 1 mmol/L MgCl2. Fraction B was subjected to DEAE ion-exchange HPLC column in the presence of EDTA. This fraction was then applied onto a DEAE ion exchange HPLC column equilibrated with 10 mmol/L Tris-HCl buffer, pH 8.0, containing 34 mmol/L octylglucoside and 1 mmol/L EDTA. By these procedures protein C, D and E were purified to apparent homogeneity as judged by SDS-PAGE. In this work, we purified the outer membrane proteins of Pseudomonas aeruginosa, and used a new technique selectively solubilizing the cytoplasmic membrane with sodium lauryl sarcosinate for isolating the outer membrane proteins of Pseudomonas aeruginosa because of its relative simplicity. PMID- 12548828 TI - [Study on the combination of isotachophoresis and high performance liquid chromatography for the separation of complex samples]. AB - A method on the combination of isotachophoresis (ITP) and high performance liquid chromatography (HPLC) for the separation and analysis of the complex samples containing proteins and metals is proposed. The instrumental system consisted of a self-made Model OCEP-1 isotachophoresis analyser, main parts of Waters high performance liquid chromatograph and a self-made Model IHI-1 interface. The interface is simple and convenient to use. The injection volume of the interface was determined accurately with a neutralization titration method. In ITP experiment the leading electrolyte was 2.5 mmol/L HCl containing 30 mmol/L of 18 crown-6; the terminating electrolyte was 5 mmol/L Tris+Cl-(pH 6.0); ITP current, 40 microA; detection of metal ions, potential gradient detector. In HPLC experiment the column was a mu-Bondapak C18, reversed-phase column, 4 mm x 200 mm, 10 microns, 12.5 nm; the mobile phase A, V(isopropanol with 5% volume fraction):V(butanol) = 2:1 containing 2.2 mmol/L HCl (pH 2.63); the mobile phase B, V(isopropanol with 50% volume fraction):V(butanol) = 2:1; low pressure gradient elution, mobile phase B changed from 0 to 70% in 17.5 minutes; flow rate, 1.0 mL/min; detection wavelength 225 nm; temperature, 25 degrees C. The experimental results showed that proteins were interfered in the determination of the metals with ITP method. It also showed that HPLC with the use of C18 column can separate the proteins but not the metal ions. By using this new combination system, a complex sample of the proteins and metal ions was injected to the HPLC, the proteins Lys, BSA and OVA were well separated in HPLC, while NH4+, K+, Na+, and Ca2+ were less retained and eluted first. Then the latter was introduced into the system of the ITP with the aid of the interface for separation. Satisfactory results were obtained. PMID- 12548829 TI - [Analysis of the isomers of dihydroxybenzenes by micellar electrokinetic capillary chromatography with on-column amperometric detection at carbon-fiber microelectrode]. AB - The effects of the concentrations of beta-cyclodextrin(CD) in electrolyte on the cyclic voltammetric properties of o-, m-, p-dihydroxybenzenes at carbon-fiber microelectrode were studied. The inclusion between o-, m-, p-dihydroxybenzenes and beta-CD was investigated based on the molecular structures and experimental results. The study showed that the order of the inclusion interaction, from strong to weak, between dihydroxybenzenes and beta-CD, was m-, o- and p dihydroxybenzenes. The effects of the concentrations of NH4Cl, SDS and beta-CD, and the pH values of the electrophoretic buffer on the migration time of o, m, p dihydroxybenzenes were investigated. The assay of o, m, p-dihydroxybenzenes by micellar electrokinetic capillary chromatography with on-column amperometric detection at carbon-fiber microelectrode was developed. The recovery ranged from 98% to 103%. PMID- 12548830 TI - [Study on the separation of Co2+ from Cu2+, Cd2+, Mn2+, Zn2+ and Fe3+ by extraction chromatography]. AB - In this paper, extraction chromatography by which Co2+ can be separated from Cu2+, Cd2+, Mn2+, Zn2+, Fe3+ is developed. The chromatographic column was 17 mm i.d. x 520 mm, containing polymers synthesized by styrene, divinylbenzene and P204 (volume fraction was 40%), with particle size of 100-200 mesh. The effects of transformation of P204 from the H(+)-form to the Na(+)-form by NaAc solution, the eluent acidity, and eluent composition were investigated. The results indicated that this extraction chromatographic method is good to separate Co2+ from Cu2+, Cd2+, Mn2+, Zn2+ and Fe3+, and the condition of separation is simple and convenient. When the P204 is transformed from the H(+)-form to the Na(+)-form by pH 4.0 NaAc (0.5 mol/L), and the column is eluted with pH 2.5 ClCHCOOH ClCHCOONa (0.1 mol/L), the separation can be completed effectively. PMID- 12548831 TI - [Investigation of association parameters of basic drugs to human serum albumin by capillary zone electrophoresis/frontal analysis]. AB - A CZE/FA method was applied to the study of association parameters of basic drugs, verapamil (VER) and propranolol (PRO), to human serum albumin (HSA) under drug-HSA association equilibria. After the direct hydrodynamic injection of a drug-HSA mixed solution into a coated capillary (effective length, 32 cm; 50 microns i.d.), the basic drug is eluted as a zonal peak with a plateau region under condition of phosphate buffer (pH 7.4, ionic strength = 0.17) at 10 kV running voltage. The unbound concentration of drug can be determined from the plateau peak height with good linear response, r = 0.999 for both VER(n = 6) and PRO(n = 8), respectively. Six VER-HSA solutions (containing 50-175 mumol/L of VER and 450-325 mumol/L of HSA) and eight PRO-HSA solutions (containing 50-400 mumol/L of VER and 450-100 mumol/L of HSA) were investigated. Three analytical methods were used for the present CZE/FA system including Scatchard, Klotz and non-linear regression analyses. The estimated association parameters were as follows: the numbers of HSA binding site n for VER were over 1.21(0.04) 1.30(0.09), binding constant, K, was between 1,690(130) and 1,790(70). Whereas for PRO, n = 0.54(0.04)-0.62(0.03), and K = 1,390(70)-1,690(180). The data deduced from these three analyses agreed well with each other. PMID- 12548832 TI - [Analysis of rice leaf proteins induced by Magnaporthe grisae with capillary zone electrophoresis]. AB - Intercellular and intracellular proteins which were extracted from rice seedling leaves induced by Magnaporthe grisae, as well as those of control, were analysed by capillary zone electrophoresis. Intercellular proteins were extracted with sterilized water, while intracellular proteins were extracted with acidic protein extraction buffer. Protein adsorption to capillary wall was suppressed by using linear polyacrylamide coated capillary (75 microns x 57 cm) instead of uncoated capillary. Separations were performed at 25 degrees C with a positive voltage of 20 kV. Samples were injected into the capillary by applying pressure (3.0 kPa) for 5 s. The running buffer consisted of 50 mmol/L phosphoric acid (pH 2.0), and detection was achieved at 200 nm with a diode array detector. Being compared with a control, a new kind of protein induced by Magnaporthe grisae was identified in rice leaf intracellular proteins with Grubbs method which is used to test outliers in mathematical statistics. PMID- 12548833 TI - [The novel copolymer coated capillary columns of electrophoresis and their applications to separation of proteins]. AB - The copolymer of acrylonitrile, methyl acrylate, hydroxy ethyl acrylate (ZB-004), the copolymer of acrylonitrile, methyl acrylate, hydroxy ethyl acrylate, acrylamide (ZB-014) and the copolymer of acrylonitrile, hydroxy ethyl acrylate (ZB-016) were coated on the inner surface of fused-silica capillaries by just filling the capillary with solutions containing these copolymers followed by flushing the capillary with nitrogen. The physically adsorbed layer can reduce both protein adsorption and electroosmotic flow in the pH range of 3-5. Electroosmotic flow decreased by raising the concentrations of the copolymers. Separation performance of ZB-004 layer is better than those of other two layers due to its low hydrophilicity, but with higher pH values, appreciable peak deformation and increase in electroosmosis were observed. The intra day and inter day migration reproducibility were investigated in terms of relative standard deviation (RSD) with four basic proteins at pH 4.0. The RSDs of the intra day migration times were less than 2%. The RSDs of the inter day migration times were less than 4%. At pH 5.0, the RSDs of the migration times in two ZB-004-coated capillaries made on two different days were less than 1%. Separation efficiencies of four basic proteins in a ZB-004-coated capillary which stored in a buffer (pH 4.0) for fifteen days after being used for 14 days decreased 15%. These coatings were stable and exhibited reproducible separations from intra day, inter day and inter column under acidic conditions. PMID- 12548834 TI - [Determination of the aconitine alkaloids in traditional Chinese medicine chuanwu and caowu by high performance capillary electrophoresis(HPCE)]. AB - A simple and rapid method for the simultaneous assay of three aconitine alkaloids (mesaconitine (MA), hypaconitine (HA) and aconitine (A) in the traditional Chinese medicines, Chuanwu and Caowu, by high performance capillary electrophoresis has been established, using tetracaine as an internal standard. The experimental conditions were as follows, the electrophoretic medium was composed of V[70 mmol/L Tris-borate (pH 8.49)]: V(methanol) = 60:40, uncoated capillary used was 50 cm x 50 microns i.d. and detection was carried out with a UV monitor at 235 nm. The separation was achieved by optimizing the cartridge temperature, the applied voltage, and the pH as well as the concentration of the buffer, and organic modifier. The calibration curve showed a good linearity in the mass concentration range of 0.0115-0.8330 g/L (r = 0.9996) for MA, 0.0089 0.6440 g/L (r = 0.9997) for HA, and 0.0083-0.6000 g/L(r = 0.9997) for A. The recoveries ranged from 93.0% to 104.0% with relative standard deviations from 0.68% to 1.7%. The total time for separation and determination was within 23 min. By means of this method, three aconitine alkaloids in Chuanwu and Caowu have been determined. PMID- 12548835 TI - [Determination of thebaine in various tissues of papaver bracteatum by gas chromatography]. AB - Papaver bracteatum is an important source of opiates because of its high thebaine content. An efficient and convenient method of determining thebaine, not only in the root, but in the more highly pigmented capsule, stem and leaf, is urgently required. We decided to examine these in detail on a wide variety of plant material and to choose an assay method which is convenient and gives reproducible results. A simple method for the quantitative extraction and analysis of thebaine in the different tissues (capsule, root, leaf and stem) of papaver bracteatum has been established. The powdered materials were extracted with methanol under ultrasonic condition. The procedure of extraction is simple, rapid and reliable. The assay method gave satisfactory reproducibility for a wide range of plant materials (coefficient of variation 2.9% to 5.4%). The linear range of response of thebaine was from 5.0 to 500.0 mg/L. The minimum limit of detection was 2 ng. The operating time of the assay is about 20 min. The presence of other alkaloids did not interfere with the assay reported here in. PMID- 12548836 TI - [The simultaneous determination of permethrin and tetramethrin in their mixed mothproofer formulation by GC]. AB - Mixed mothproofer of permethrin and tetramethrin is a new formulation for prevention cloth moth in China. A rapid and specific method is needed to simultaneously determine these two active ingredients. For this purpose, a method by GC was established with a 3% OV-101/Chromosorb WAW DMCS(180-250 microns) column at a temperature of 223 degrees C, by using di-iso-octylsebacate as internal standard. The mean coefficient of variation for permethrin and tetramethrin were 0.20% and 0.19% respectively. Linearity of detector response vs. mass concentration ranged from 0.38-2.6 g/L with r = 0.9999. PMID- 12548837 TI - [Determination of benzidine in cotton cloth by high performance liquid chromatography(HPLC)]. AB - In this paper, a method for determination of benzidine in cotton cloth by HPLC was studied. First, cotton cloth was treated by sodium dithionite at the temperature of (70 +/- 2) degrees C. On this condition, azo dyes in cotton cloth were reduced to benzidine. Then benzidine was extracted by diethyl ether in sodium hydroxide solution. Benzidine originating from cotton cloth was analysed by HPLC with a column of Hypersil ODS (5 microns, 15 cm x 4.6 mm i.d.), mobile phase of V (MeOH):V(H2O) = 75:25, flow rate of 0.6 mL/min, pressure of 17.7 MPa and detection at 254 nm. By comparison with the standard sample, the retention time of benzidine was about 2.5 min. Benzidine in three samples was determined through parallel experiment. The mass ratio of benzidine in cotton cloth was 94.01, 86.9 and 120.8 micrograms/g, and the average was 100.57 micrograms/g. The average recovery was 84.7%. The experiment expressed that benzidine originating from cotton cloth could be qualitatively and quantitatively determined by HPLC. The accuracy of the results of parallel experiments was related to the reducing process. When the reducing conditions were completely consistent, the result would be more accurate. PMID- 12548838 TI - [Determination of monensin residue in chicken by HPLC with post-column derivatization]. AB - The monensin residue was extracted from the tissue by homogenization with methanol-water and the extract was filtered and partitioned with dichloromethane. The dichloromethane extract is concentrated and clean up by passing through a silica gel cartridge. The analyte on the cartridge is then eluted with dichloromethane-methanol. The eluate is collected and evaporated to dryness. The residue is dissolved and made to a definite volume with 1 mL methanol and the solution is used for post-column derivatization-HPLC determination. Monensin is separated on mu-Bondapak C18 column (3.9 mm i.d. x 300 mm) with methanol-water phosphoric acid as a mobile phase and the flow rate was 0.7 mL/min. The eluted monensin was reacted with vaniline under acidic and heated condition in post column derivatization system then detected at 520 nm and quantitated by external standard method. The derivatization reagent consisted of 20 mL concentrated sulfuric acid, 950 mL methanol and 30 g vaniline. The flow rate was 0.7 mL/min. The reactor was a stainless steel coil (300 cm x 1 mm i.d.) set in a 90 degrees C oven. The response values was linear between 20-200 ng. The recovery was 88.1% 101.3%. The coefficient of variation was 0.1%-0.73%. PMID- 12548839 TI - [Simultaneous analysis of histamine and noradrenaline in human plasma by high performance liquid chromatography/electrochemical detection]. AB - This paper reports a reversed-phase high performance liquid chromatographic method for the simultaneous separation and determination of histamine(Hist) and noradrenaline(NA) in human plasma by means of pre-column derivatization of Hist and NA with OPA in the presence of 2-mercaptoethanol. The analysis was carried out with Partisil ODS column, V[sodium acetate buffer solution(pH 4.90)]:V(acetonitrile) = 7:3 as mobile phase and 3-methylhistamine as internal standard. The detector potential was +0.7 V for both compounds to keep minimum background noise. A linear relationship between the electrochemical signal and the compound mass concentrations was obtained. The minimum detectable limit was 15 ng/L (S/N = 3). An example of application of the method to the determination of Hist and NA in human plasma is presented. PMID- 12548840 TI - [Analysis of herbicide-difenzoquat by reversed-phase high performance liquid chromatography (RP-HPLC)]. AB - An RP-HPLC method for the qualitative determination of herbicide difenzoquat was developed. Operating conditions were UV-254 nm detection, Selectosil 5 C18 column 250 mm x 4.6 mm i.d., and V(methanol):V(water) = 60:40 (pH3, adjusted by H3PO4) as mobile phase. The coefficient of variation of this method was 0.41% with recoveries from 98.71% to 100.36%. The retention time was about 6.6 min. The coefficient of linear correlation was 0.9999. The method is very effective for the quality control of SP (water soluble power) products of this pesticide. PMID- 12548841 TI - [An improvement on determination of the related substances in vindesine sulfate and its injection]. AB - A new method for the analysis of related substances in vindesine sulfate and its injection to improve the ability of separation by RP-HPLC was established. The separation was achieved by applying a reversed-phase C18 column (150 mm x 4.6 mm i.d.) and a mixture of V(MeOH):V(buffer solution) = 70:30 as mobile phase. The buffer solution was an aqueous solution of volume fraction 1.5% diethylamine adjusted to pH 7.5 with phosphoric acid. The UV-detector was set at 267 nm. Four related substances were separated from vindesine within thirteen minutes. The tailing factor of the peak of vindesine is 1.00-1.05. The results showed that this method is more efficient for separating the related substances in vindesine sulfate than the conventional method. PMID- 12548842 TI - [Analysis of the composition in fructooligosaccharide by high performance liquid chromatography]. AB - In this paper, an analytical method for determination of fructooligosaccharide by high performance liquid chromatography(HPLC) is presented. The HPLC conditions are mu-Bondapak C18 column, 300 mm x 3.9 mm i.d., water as mobile phase and refractive index detector. Monosaccharide, disaccharide, trisaccharide, tetrasaccharide, pentasaccharide, hexasaccharide etc. can be simultaneously determined in a single injection. The method has been used to determine the fructooligosaccharides in "Boai biogen". The results are satisfactory. The CVs were 0.55%-1.1% and correlation coefficients were 0.9992-1.000. The lowest detectable limit for fructooligosaccharide was in microgram level. PMID- 12548843 TI - [Determination of glycyrrhizic acid in glycyrrhiza preparations with capillary electrophoresis and high performance liquid chromatography]. AB - High performance liquid chromatography (HPLC), capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography(MECC) have been used to determine glycyrrhizic acid in glycyrrhiza preparations. By HPLC, mobile phase was V(methanol):V(water):V(acetic acid) = 75:23.5:1.5. By CZE, experiment was performed with 15 kV power, 0.075 mm i.d. x 800 mm fused-silica capillary column and UV detector. Samples were injected into the capillary by electromigration injection for 20 s. Absorbance detection was at 254 nm. The running buffer was made up of 0.02 mol/L dipotassium hydrogenphosphate and borax (pH 9.0). By MECC, the running buffer was made up of 0.025 mol/L sodium dodecyl sulfate (SDS), 0.02 mol/L dipotassium hydrogenphosphate and borax (pH 9.0). Each new capillary was washed with 0.1 mol/L NaOH, deionized water and buffer, each for 3 min, before use. Comparison of the results of analysis with HPLC, CZE and MECC has been made. It was found that result of MECC was very close to that of HPLC. MECC has been satisfactorily applied to plant drug analysis. PMID- 12548844 TI - [Rapid method for determination of chlordimeform residue in honey]. AB - In the present work. TLC was used as a rapid method for determination of chlordimeform in honey. Twenty gram of sample was extracted by n-hexane in alkali solution, re-extracted by acidic aqueous solution, then hydrolyzed in NaOH solution for 10 min on boiling water bath. The hydrolyzed products were extracted by methylene chloride. After passing through the Na2SO4 column for dehydration, the methylene chloride should be evaporated to less than 70 degrees C to 0.5 mL spot 50 microL sample solution on the silica gel plate, develop the plate about 8 cm from the surface of developing solvent V(benzene):V(chloroform):V(ethyl acetate) = 5:5:1. The plate was kept in the atmosphere (NaNO2 + HCl) for 10 min and 5% N-(1-naphthyl) ethylendiamine dihydrochloride was sprayed on it. Put it into a dark room for 20 min and then observe and estimate the amount of chlordimeform in the sample. The limit of determination is 10 ng. The experiment could be completed in 3 h for a sample without large equipment and special reagent. PMID- 12548845 TI - [Determination of dicarboxylic acids in a new compound liquid fertilizer by gas chromatography]. AB - A method for determination of succinic acid, glutaric acid and adipic acid in a new compound liquid fertilizer by gas chromatography is described. The sample after dried at 80 degrees C was extracted by ethyl alcohol, and then esterified in the presence of BF3. The esterified derivatives were injected into a gas chromatograph with a 5% SE-30 packed column, programmed from 140 degrees C (held for 1 min) to 160 degrees C (held for 6 min) at a rate of 6 degrees C/min. Internal standard was diethyl malonate. Calibration curves of the three diethyl esters were linear in the ranges of 2.3-9.3 micrograms, 10.8-43.2 micrograms and 2.8-11.0 micrograms respectively. PMID- 12548846 TI - [The effect of heat shock before rat partial hepatectomy on HSC70/HSP68 expression and phosphatase activities]. AB - The contribution and content of the continuous heat shock protein 70/induced heat shock protein 68 (HSC70/HSP68), the contribution, variety and activity of acid phosphatases (ACP) and alkaline phosphatases (AKP) had been analysed qualitatively and quantitatively during the liver regeneration after 2/3 hepatectomy (PH) and HS (heat shock at 46 degrees C for 30 min, recovery for 8 h), which were compared with the results only by HS and only by PH. It was shown that the three kinds of treatment all can increase the activity of ACP, AKP and the expression of HSC70/HSP68, but with different change pattern. A further analysis show that after HS-PH the enhanced activity of ACP is related with that of 140 kD phosphatases, the enhanced activity of AKP is associated with that of 140 kD and 160-180 kD phosphatases. It can be reckoned from the results that ACP, AKP and HSC70/HSP68 all act on the heat shock response of hepatocyte and liver regeneration, and may take part in signal transduction in these processes, but ACP may play a dominant role in the start of hepatocyte multiplication, AKP and HSC70/HSP68 may play a dominant role in cytokineses. PMID- 12548847 TI - [The apoptosis of K562 cells induced by IL-3 and hydroxyurea cooperatively]. AB - The effect of IL-3 and hydroxyurea on human erythroleukemia cell line (K562 cells) was demonstrated by using the electro-microscopy and flow cytometry. Our data showed that neither IL-3 nor hydroxyurea could induce the apoptosis of K562 cells alone. However, the IL-3 and hydroxyurea could induce the apoptosis of K562 cells cooperatively. Analysis with flow cytometry showed that the percentage of apoptotic cells was about 31.90% after K562 cells were induced by IL-3 and hydroxyurea cooperatively for 5 days, and the sub-G1 peak (apoptotic peak) was detected in the induced K562 cells. Meanwhile, the percentage of S-phase in the IL-3 and hydroxyurea induced K562 cells was increased, and the proliferation of the induced K562 cells was inhibited significantly. Furthermore, the IL-3 and hydroxyurea induced K562 cells showed chromatin condensation with regular crescents at the nuclear edges and apoptotic bodies. It suggested that IL-3 could enhance the sensitivity of K562 cells to hydroxyurea and the apoptosis of K562 cells could be induced by IL-3 and hydroxyurea cooperatively. PMID- 12548848 TI - [The inhibitory effect of asODNs on the invasion of colorectal cancer cell line CCL229]. AB - To study the inhibitory effect of antisense oligodeoxynucleotide (asODNs) on colrectal cancer cell line CCL229 invasion in vitro. A 15-mer asODNs targeted against the translation start site of UPAR (urokinase-type plasminogen activator receptor) mRNA were introduced into CCL229 cells by lipid-mediated DNA transfection and the variation of the levels of uPAR mRNA, uPAR antigen expression of the levels of uPAR mRNA, uPAR antigen expression on the cell sruface and invasion properties were observed by reverse transcription-polymerase chain reaction (RT-PCR), flowcytometry(FCM) and aminion invasion assay, the morphological feature of the cell after asODNs treatment was observed by scanning electron microscope(SEM). The results indicate (1) the uPAR/beta-actin ratio was 0.44 +/- 0.02 for the asODNs treated cells, which is significantly lower compared with the control and rONDs treated cells (0.81 +/- 0.01 and 0.750 +/- 0.13 respectively, P < 0.01), (2) the mean fluorescence index of uPAR combined with uPA and the whole uPAR on surface were 0.20 +/- 0.07 and 0.59 +/- 0.09 respectively for asODNs treated cells, which is significantly lower compared with control cells (0.72 +/- 0.12 and 2.21 +/- 0.36 respectively, P < 0.05, P < 0.01); (3) the number of cells migrated the aminion (25 +/- 4, 44 +/- 5 for the control cells) obviously decreased after a-sODNs treatment, (12 +/- 2, 20 +/- 3, P < 0.05); (4) the filopodia and microspikes on the CCL 229 cell surface were decreased after asODNs treatment. The conclusion is that the expression of uPAR on the surface of CCL229 cell surface is responsible for invasity; the inhibitory effect of uPAR as ODNs were highly significant and this method may be of potential clinical interest in gene therapy of cancer. PMID- 12548849 TI - [Cloning and identification of a new telomeric-associated zinc finger protein cDNA]. AB - In order to isolate novel genes related to early human embryo development and differentiation, a directional cDNA library was constructed from 3-week-old human embryo. Single-pass DNA sequence analysis was used to sequence 47 randomly picked low-abundance cDNA clones. This approach led us to select a clone, L30, showing significant homology with the telomeric-associated DNA and zinc finger protein genes. It is about 3.8 kb in length and contains an open reading frame of notable length within 5'-region, and a tailing signal of AAUAAA and poly (A+) with 39 A in 3'-region. The gene was transcribed in human embryo by Northern blot hybridization and assigned to human chromosome 12 by in situ hybridization. PMID- 12548850 TI - [The capability of IL-1 in the induction of chemokine production by murine thymic epithelial cell (MTEC1) clones]. AB - To analyze the capability of IL-1 and IL-6 in the induction of chemokine (CF) production by mouse thymus epithelial cell (MTEC1) clones, MTEC1 cells were cloned through one cell culture and individual cell clones were established in long term culture referred to as MTEC1-DW clones. The constitutive production of IL-1, IL-6 and CF by MTEC1-DW clones was evaluated and the patterns of the cytokine production determined. Addition of exogenous IL-1 or IL-6 or both to the cultures of those MTEC1-DW clones that are unable to produce CF, and incubated for 2 days, then, to assess the chemotactic activity in the cell culture supernatants (SNs). In the opposite, addition of anti-IL-1 mAb(s) to the cultures of those MTEC1-DW clones that can produce IL-1 and CF to neutralize secreted IL-1 then, to test chemotactic activity in the SNs after 2-day incubation. The results showed that in the MTEC1-DW clones which were unable to constitutively produce IL 1 or CF, addition of IL-1 could induce these cloned cells to produce CF with high chemotactic activity. By constrast, addition of anti-IL-1 mAb(s) to those MTEC1 DW clones that constitutively produce IL-1 and CF could significantly inhibit them to produce CF. IL-6 only exhibited weak activity in the induction of CF production by those cloned cells. Therefore, in the cytokine network regulation, CF production is mainly induced by endogenously produced IL-1 in MTEC1 cells. PMID- 12548851 TI - [Studies on the localization of NPY-like immuno-reactivity in the nervous system during development of Cynops orientalis]. AB - By immuno-cytochemical method the localization of neuropeptide Y (NPY) in the nervous system during embryonic development of Cynops orientalis was studied. The results revealed that NPY was first localized in the peripheral nervous system (late tail-bud stage), and later appeared in the central nervous system (larval stage) where it appeared with the appearance of glial cells. Very probably with the migration of neural crest cells NPY appeared first in the peripheral nervous system and then distributed to the central nervous system. PMID- 12548852 TI - [Cloning and analyzing of members of excitatory amino acid transporter family from neonatal mouse brain]. AB - Excitatory amino acid transporter family (EAAT) contains several structure related membrane proteins. They are essential for the removal of glutamate released from pre-synaptic terminal to terminate its action of synaptic transduction and maintaining the normal concentration of neurotransmitters in nerve system. To study these proteins in single animal model, we cloned several members of EAAT family, named mGLAST-1, mGLT-1, mEAAC1 and mASCT1, from a neonatal mouse brain cDNA library. The cDNA sequence of mASCT1 was firstly reported in mouse, it is composed of 3787 bp which has an open reading frame (ORF) encoding a protein of 532 amino acid residues. The mASCT1 protein was expressed in Xenopus oocyte and the function was characterized by 3H-Ser uptaking. The homology between human ASCT1 and mouse ASCT1 is 89.3%. The DNA sequence data shows the variance in length and composition exists in the sequence of 5'UTR and 3'UTR of mRNA in the family members of EAAT. This phenomenon may indicate a post-transcription regulation mechanism might exist in the gene expression of mouse EAAT family members. PMID- 12548853 TI - [Transformation of sainfoin by Agrobacterium rhizogenes LBA9402 Bin19 and regeneration of transgenic plants]. AB - Hypocotyl segments of Onobrychis viciaefolia were transformed by Agrobacterium rhizogenes LBA9402 which harboured pBin19 and pRi1855. Seedling age and preculture time of hypocotyl segments influenced the transformation frequency. Paper electrophoresis revealed that 70% of single hairy root cultures could synthesize agropine. Calli were induced from hairy root segments on MS medium containing 0-9.05 mumol/L 2,4-D and 0-2.22 mumol/L 6-BA at first, then they were transferred onto MS0 medium without kanamycin for regeneration. Constitution and concentration of phytohormones in callus induction media affected subsequent regeneration of calluses on MS0 medium remarkably. Regeneration frequency and shoot number per callus declined when 2,4-D concentration in callus induction media increased from 4.52 to 9.05 mumol/L, while they ascended when 6-BA in callus induction media increased from 0 to 2.22 mumol/L. On MS medium supplemented with 4.52 mumol/L 2,4-D and 2.22 mumol/L 6-BA, only 14.2% hairy root segments could produce calluses, but the regeneration frequency reached 58.1% and the shoot number per callus was 37.2. In 32 analysed plants regenerated from 8 kanamycin resistant hairy root lines, 25 were nptII positive and showed different copy numbers. PMID- 12548854 TI - [Factors on somatic embryogenesis of soybean elite cultivars and plant regeneration]. AB - Factors on in vitro somatic embryogenesis of soybean (three elite cultivars) were studied using cotyledons of 3.0-6.0 mm immature seed as explants. Not only the kinds, concentrations and combinations of plant growth regulatory substances but also immature embryo length and inoculum density have main effects on the approaches of embryogenesis. The results of two-factors analysis of variance experiments showed that immature embryo length, plant growth substance concentration and basic medium type have very significant effects on the frequency of embryogenic response, furthermore, interactions exist between the former two factors and are just very significant(at 1% level). The best combinations between 2,4-D concentration and cotyledon length are 10 mg/L 2,4-D & 4.0 mm immature embryos, 20-40 mg/L 2,4-D & 5.0 mm immature embryo. Under these combinations, the salt composition of E1 are very significantly better than that of MS. In conclusion, in the regeneration system established by us the frequency of somatic embryogenesis from the soybean immature cotyledons is greater than 50% and the frequency of conversion of normal (not fused) somatic embryos is about 52.9%-62.6%. PMID- 12548855 TI - [Isolation of a gene related to trichosanthin-induced apoptosis (GRETA)]. AB - Disturbance of the apoptosis-related signaling pathway is regarded as one of the critical factors for tumorigenesis. Isolation of the genes involved in the process of apoptosis would be thereby helpful to explore the mechanism of tumor transformation and to develop novel therapeutic approaches. Here we report a gene fragment GRETA, the gene related to trichosanthin-induced apoptosis, isolated from a leukemia cell line U937 undergoing apoptosis induced by a plant protein Trichosanhin (TCS). A 293bp segment of GRETA was revealed to be 78.3% homologous to Bruton's tyrosine kinase at nucleic acid level. And Northern blot analysis showed that three messengers of RNA with the size of about 0.8-kb, 2.0-kb and 7.0 kb, respectively, were detected in TCS-untreated U937 cells when probed with GRETA, but there were only 0.8-kb and 2.0-kb transcripts appeared in apoptotic U937 cells. In addition, the abundance of each transcript changed apparently. The 0.8-kb transcript, for example, was the main band in Northern analysis in apoptotic U937 cells while was only detected marginally in TCS-untreated cells. These data suggested a possible relationship between the alternative splicing patterns of GRETA and the apoptosis. PMID- 12548856 TI - [Factors influence on transformation by particle bombardment in Indica rice]. AB - Four factors influence on transformation of indica rice, which were high osmotic treatment; different explant as the target tissue; pressure of rupture disk and quantity of plasmid DNA, were investigated in this experiment. High osmotic treatment of target tissue prior to and after bombardment increased 3.2-fold for Gus transient expression than control. The best treatment of high osmotic was that the target tissues were kept in the target-bed medium which contained 0.4 0.6 mol/L sorbitol and manitol each for 4 h prior to bombardment and for 16 h after bombardment. Four explants: scutellum from mature seed, young panicle, embryogenic callus and suspension cells of indica rice were tested as target explant by particle bombardment. The results of Gus transient showed that the highest expression was scutellum and for other three explants, the order from high to low was young panicle, embryogenic callus and suspension cell. Transgenic plants were obtained from all of the explants except young panicle. For the pressure of rupture disk on transformation, 1100 psi or 1300 psi of the pressure of rupture disk were best one for the transformation and higher than 1300 psi could damage the target tissue which become black and died in the following culture duration. For the quantity of plasmid DNA, the results showed that 0.83 microgram of plasmid DNA per bombardment was preferred for the transformation of indica rice. PMID- 12548857 TI - [Ultrastructural observation of E. coli K12 treated with antibacterial peptide CM4]. AB - The effect of antibacterial peptide CM4 of Bombyx mori against E. coli K12 was investigated using scanning electron microscopy(SEM) and transmission electron microscopy (TEM). The ultrastructural changes of E. coli K12 were observed by the challenge of the purified antibacterial peptide CM4. The results showed that the antibacterial peptide caused a series of pathological changes on E. coli. SEM and TEM revealed aggregates of bacteria and SEM revealed wrinkled bacterial surfaces in the early stage. Thereafter, plasmolysis was observed with irregular holes appearing in the two ends of bacteria and the cytoplasmic contents of the cells leaking out. Finally, bacteria became empty vesicles and disintegrated into small fragments subsequently. Comparatively, the bacterial membrane was normal and the bacterial structure remained intact in the control group. PMID- 12548858 TI - [Induced-differentiation and cytotoxicity of isoverbascoside on HL-60 cells]. AB - HL-60 cells were treated by isoverbascoside with different time and different concentrations in vitro. The differentiation of HL-60 cells was evaluated by light and electron microscopy to observe morphological changes, by chemiluminence to detect phagocytosis and by tumorigenesis in nude mice to determine malignancy. The cytotoxical effect of isoverbascoside on HL-60 cells was examined by trypan blue excluding staining and electron microscopy. The influence of isoverbascoside on cell cycle was measured by flow cytometry. Granular differentiation of HL-60 cells was induced by isoverbascoside at 20-25 mumol/L within 1-3 days as the results of morphological changes, enhancement of phagocytosis and decreasing of tumorigenesis. Strong cytotoxicity was evidenced in HL-60 cells treated by isoverbascoside at 30-35 mumol/L. HL-60 cells treated by isoverbascoside at 20 mumol/L were delayed at G1 phase at 12 hours and G2/M phase at 72 hours. PMID- 12548859 TI - [Construction of human pLXSN-CD80 retrovirus vector and expression in CHO and PA317 cells]. AB - CD80, a molecule on the antigen presenting cells, provide costimulation signals for T cell activation and play a key role in tumor immune. Based on our previous work of human CD80 full length cDNA cloning, a retrovirus expression vector pLXSN CD80 was constructed. CD80 expression cells were selected by G418 from pLXSN-CD80 transfected PA317 and CHO cells by calcium phosphate. Expression, distribution and molecular weight (MW) of CD80 were measured by RIA, FACs and western blot. pLXSN-CD80 transfected CHO cells expressed relatively high level of CD80 protein (approximately the same as Raji cells) with an apparent MW of 40 kD. In the presence of G418 or not, pLXSN-CD80 transfected PA317 and CHO cells maintained CD80 expression for five months of passage. The results indicate that our construct is potent for experimental use in gene therapy. PMID- 12548860 TI - [Stable expression of recombinant inducible nitric oxide synthase in NG108-15 cells and its biological characterization]. AB - The neuroblastoma x glioma NG108-15 cells were transfected with recombinant eukarytic expression plasmid pCMViNOS containing the full-length cDNA encoding inducible nitric oxide synthase (iNOS). A lot of G418-resistant clones were screened at 600 micrograms/ml of geneticin. In the 2# clone expressing iNOS gene, iNOS catalytic activity in the cytosol fraction displayed to have an increasing trend, accompanying with the accumulation of NO2- content in the supernantant of cultured cells and the intracellular cGMP concentration, which suggested that NO cGMP signal pathway was mediated by the expression of iNOS gene and blocked by NG nitro-L-arginine (L-NNA) and methylene blue (MB). Activity of iNOS was concentration-dependently inhibited by NOS inhibitors such as L-NNA and aminoguanidine. The result of measurement of NADPH diaphorase activity and immunocytochemical staining showed that localization of the function expression of iNOS protein mainly existed in the cytoplasm of NG108-15 cells transfected with pCMViNOS. Furthermore, the chromosomal integration, transcript and protein translation of foreign iNOS gene were identified by Southern hybridization, RT PCR and Western blot, respectively. The results indicated that iNOS gene transfected cells had mRNA transcription and specific protein expression at high level. Given the above results, the engineering cell line with stable expression of iNOS gene was successfully established. The new neuronal cell line may serve as a source of iNOS and provide a useful cell model for studying iNOS biological function and developing novel iNOS-selective inhibitors. PMID- 12548861 TI - Effects of ginsenosides on myocardial reperfusion arrhythmia and lipid superoxidation in high cholesterol diet rats. AB - To explore the effects of GSL on myocardial reperfusion arrhythmia and lipid superoxidation in high cholesterol diet rats. Hyperlipidemia model was set up with administered high cholesterol emulsion 15 ml/kg to rats orally for 14 days. In GSL group, rats were given GSL i.p. 75 mg/kg simultaneously when administered high cholesterol emulsion. The experiment of myocardial ischemia reperfusion was performed on all rats. The results showed: (1) After administration of high cholesterol emulsion to rats orally for 14 days, hyperlipidemia model was set up successfully, simultaneously treatment with GSL. It lowered serum lipid; (2) In hyperlipidemia state, serum MDA increased (p < 0.01, SOD and NO decreased markedly (p < 0.01 and p < 0.05 respectively) after 2 h of myocardial reperfusion; the rate of reperfusion arrhythmia (RPAr) increased within 10 min of reperfusion, four out of nine rats died of ventricular fibrillation (VF); and (3) GSL decreased MDA, increased SOD and NO after 2 h of myocardial reperfusion. All changes were significant (p < 0.01); the rate of RPAr decreased, no VF occurred and all rats survived. Hyperlipidemia aggravated myocardial ischemia reperfusion injury and increased the incidence of RPAr. The results suggested that GSL reduced myocardial ischemia reperfusion injury and RPAr in high cholesterol diet state through antiperoxidating and inducing the production of NO. PMID- 12548862 TI - [Characterization of expression of the apoptosis associated PF18-3 molecule in thymocyte apoptosis following ConA activation]. AB - The expression pattern of McAb PF18-3 recognized molecule in activation induced thymocyte apoptosis was analysed. Results indicated that following activation by ConA, thymocytes underwent activation induced apoptosis identified by delayed production of DNA ladder and TUNEL positive staining of thymocytes in late stage of activation. PF18-3 molecule was found to express specifically in subset of apoptotic thymocytes with diploid. Kinetic comparison of expression between PF18 3 molecule and Fas or translocated membrane phospholipid suggested that PF18-3 molecule is different from them and likely to be a novel molecule related to thymocyte apoptosis. PMID- 12548863 TI - [The relationship between fMLP induced neutrophil respiratory burst and the apoptosis of neutrophil]. AB - The relationship between apoptosis of neutrophils and the change of their intracellular free Ca2+ concentration [Ca2+]i was studied. FMLP and A23187 were used to elevate the [Ca2+]i while BAPTA was used to deplete it. Fluorescence microscope, flow cytometry and gel electrophoresis were used to study the percentage of cell apoptosis and the change of f-actin during apoptosis. The results showed that the apoptosis was obviously inhibited by fMLP and A23187, while accelerated by BAPTA. The detection of f-actin showed that the f-actin depolymerized obviously during apoptosis. The elevation of [Ca2+]i inhibit the actin depolymerization while depletion of [Ca2+]i accelerated it. This result indicated that the apoptosis of neutrophil was obviously inhibited by [Ca2+]i elevation but accelerated by [Ca2+]i depletion. PMID- 12548864 TI - [Effects of inhibition of PKC alpha on breast cancer cell phenotype and expressions of cyclin E and CDK2]. AB - Human breast cancer cell line Bcap-37 was stably transfected with the plasmid expressing antisense PKC alpha RNA, and cells, in which PKC alpha was inhibited due to antisense PKC alpha RNA, were isolated. Changes in serum-dependent growth in cell culture, cell clonogenicity in soft agar and growth in nude mice were tested, and the expressions of cyclin E and CDK2 were analyzed. After PKC alpha was inhibited, the cells showed that serum-dependent growth and anchorage dependent growth enhanced, tumorigenicity in nude mice decreased. The results suggest that less aggressive breast cancer phenotypes may be induced by inhibition of PKC alpha. Levels of cyclin E and CDK2 mRNA in cells with antisense PKC alpha RNA were lower than those in control cell. These indicate that signal transduction system with PKC alpha is closely related to cell cycle control system with cyclin/CDK in the functions. PMID- 12548866 TI - [Agrobacterium tumefaciens mediated maize transformation]. AB - Several maize inbreds were transformed with Agrobacterium tumefaciens EHA101 (pGIH). Transgenic maize plants were obtained. Frequency of transformation of maize inbred Suyu No. 1 can reach 8.1%. Results of PCR and Southern blot analysis proved that T-DNA was stably integrated into the genome of maize. Staining with X gluc confirmed the expression of GUS gene in maize cells. The band amplified by inverse PCR showed that the copy number of transgene in three transformants was single. After long term of subculture, some hygromycin resistant calli lost their regeneration ability. Although Southern blot probed the integration of gusA gene in their genome, GUS activity cannot be detected in those calli. Southern blot analysis of HpaII digest DNA showed that transgenic gusA gene was highly methylated. PMID- 12548865 TI - [The gene expression of some cytokines and collagen proteins in rat bone tissue is related to estradiol (E2) and age]. AB - 30 female SD rats (3 months old) are equally divided into three groups: ovariectomy (OVX) rats, sham-operated (SHO) rats and 17 beta estradiol (E2) treated OVX rats. For each group, mRNA was isolated from long bone at one month and three months after surgery, respectively. mRNA was reverse transcribed into single strand cDNA and then used as a probe hybridizing to the DNA fragments of col I alpha(1), col I alpha(2), col III, col V, fibronectin, IL-1, IL-6, TGF beta, LIF, TNF-alpha, TNF-beta by reverse northern and dot blot hybrization. The housekeeping gene, gapdh, was used as an internal control. The results show that in bone of rat, the stable expression of col I alpha (1), col I alpha(2) and col III are related to age not ovariectomy, while supplement with E2 can inhibit the expression of col III and col I alpha(2) completely. The expression of col V, IL 1, IL-6 can be inhibited by estrogen and recovered by removal of estrogen by OVX, then addition of E2 decreased it to the normal level. The expression of TGF-beta is also inhibited by estrogen. It increased during one month after overiectomy, and partially decreased in E2 complemented rat. Three months after surgery, the level of increasing and decreasing is less evident as two months ago. It seems that in young SD rat, the expression of TGF-beta is related to both estrogen and age. PMID- 12548867 TI - [Study on factors influencing Agrobacterium-mediated cassava transformation]. AB - Factors influencing agrobacterium-mediated cassava transformation were investigated. Among the four Agrobacterium strains tested, LBA 4404 (pTOK 233) and LBA 4404 (pBin9GusInt) gave higher transient expression than C 58 C1 (pIG121Hm) and EHA 105 (pBin9Husint). Pretreatment of explants by bombardment or vaccum had no significant effect on transient expression while preinduction of Agrobacterium with acetosyringone showed better effects, and preculture of explants showed worse effects. All the cultivars tested were susceptible to Agrobacterium infection, while the types of explants and the physiological state of the explants had a strong influence on the transient expression efficiency. The 15-day-old somatic cotyledons and the fully expanded leaves from in vitro plantlets were the most susceptible to Agrobacterium infection. The results also showed that all the four selective reagents (hygromycin, geneticin, PPT, and kanamycin) synchronously suppressed the growth of callus, shoot organogenesis and shoot rooting in a dose dependent manner. PMID- 12548869 TI - [Promotion of transformation frequency of soybean (Glycine max L.) protoplasts using poly-L-ornithine]. AB - The foreign Bt gene was transferred into protoplasts of soybean using PEG and PLO methods, respectively. The result indicated that the transformation frequency of PLO method was about 0.1% higher than PEG method. The PCR and Southern blotting analysis of the regeneration plants confirmed the integration of foreign gene into the genome of soybean. PMID- 12548868 TI - Regeneration of Astragalus adsurgens via somatic embryogenesis from cell suspension protoplasts. AB - Protoplasts from 4-day-old embryogenic cell suspension cultures of Astragalus adsurgens, when cultured in KM8P medium which ammonium concentration was reduced to 2.5 mmol/L and supplemented with 0.5 mg/L NAA, 1.0 mg/L 2, 4-D, 0.7 mg/L BA and 0.4 mol/L glucose, underwent cell sustained divisions and formed cell colonies at a frequency of 16%-20%. Preplasmolysis or low temperature treatment of suspension cells prior to enzyme incubation enhanced colony formation. Following proliferation on MS medium containing 1.0 mg/L 2, 4-D and 0.5 mg/L BA, cell colonies were cultured on MS medium containing 0.1 mg/L NAA and 1.0 mg/L BA, where approximately 40% of colonies produced somatic embryos ranging in number from 20 to 40 per colony. No significant decrease was found in the potential of somatic embryogenesis when protoplast colonies were obtained from long-term cell suspensions. On hormone-free 1/2 MS medium, somatic embryos developed into intact plants, which showed normal morphology and stable chromosome number. PMID- 12548870 TI - [Phylogenetic analysis of two rhizobia new groups]. AB - On the basis of numerical taxonomy, SDS-PAGE whole-cell protein analysis, 16S rDNA PCR-RFLP and DNA-DNA hybridization, the full length of 16SrDNA of two strains (XJ96060, XJ96408) were sequenced and compared with the 16S rDNA sequences of other members of the alpha 2 subclass of the Proteobacteria available from GenBank. An unrooted tree was produced to determine the phylogenetic relationships of two groups. The results showed that all strains were clustered into 6 branches, they were Sinorhizobium, Mesorhizobium, Agrobacterium-Rhizobium, Bradyrhizobium. Rhizobium, Azorhizobium. Group 1, 2 fell into Sinorhizobium branch. PMID- 12548871 TI - [Reclassification of Saccharomyces strains by comparative electrophoretic karyotyping]. AB - The strains of Saccharomyces Meyen ex Reess preserved in China General Microbiological Culture Collection Center (CGMCC) were recharacterized and reidentified according to recent taxonomic improvement of the genus. The strains AS 2.100 (originally classified in S. cerevisiae), AS 2.1158(from former USSR and originally classified in S. exiguus) and AS 2.1555(from Australia and originally classified in S. uvarum) were found to be different from the standard descriptions of the species concerned in some physiological properties. Comparative CHEF electrophoretic karyotype analysis showed that the chromosomal DNA banding pattern of AS 2.100 was similar to that of the type strain of S. bayanus, while the electrophoretic karyotypes of AS 2.1158 and AS 2.1555 were similar to those of the type and authentic strains of S. cerevisiae. Therefore, AS 2.100 was reidentified as S. bayanus, and AS 2.1158 and AS 2.1555 were reclassified in the species S. cerevisiae. PMID- 12548872 TI - [Characterization and phylogenic analysis on a new isolate of genus Rhodocista]. AB - A strain of purple non sulfur bacteria was isolated from waste water plant in Beijing. It is phototaxic, and can converted the vibrioid cells into thick-wall cyst. Internal synthetic membrane is present as lamellae. It has a low absorption maxima at 798 nm. and require thiamine and vitamin B12 as growth factors. It can use succinate as carbon and energy source, but glucose can not be used. It can not grow in 3% NaCl and had Q-9 as major quinone. The 16S rRNA gene was amplified, cloned and sequenced. A phylogenic tree was constructed on the basis of the 16S rDNA sequences. It showed that the previously known member of the genus Rhodocista, Rc. centenaria, is the nearest neighbor to strain 3-p. The level of binary sequence similarity between Rc. centenaria and strain 3-p of 95%, and their phenotypic differences and genetic DNA-DNA relatedness of 56%, shows they are different species of Rhodocista. A new species name, Rhodocista pekinense, was proposed for strain 3-p. PMID- 12548873 TI - [A Carnobacterium-like organism isolated form a patient with multiple bacterial synergistic gangrene]. AB - An atypical lactic-acid producing gram positive rod Y6 strain was studied in this report, which was isolated form clinic sample of a patient with multiple bacterial synergistic gangrene, and could not be identified by routine method. A 1.5 kb of 16S rDNA of Y6 strain was synthesized and sequenced. Comparative 16S rDNA sequence analyses revealed that stain Y6 is most closely related to the genus of Carnobacterium. The overall similarity value between Y6 strain and Carnobacterium species are 93% to 97%. The signature nucleotides in 16S rDNA primary sequence of strain Y6 and that of genus of Carnobacterium were identical. The biological features of Y6 strain are very similar to that of Carnobacterium, such as lactic acid as main end product of in PYG broth, no gas produced form fermentation of glucose, catalase negative, no motility. Data suggested that Y6 strain is very similar to the genus of Carnobacterium, of which no strain has been isolated form clinical sample so far. Based on the results obtained, we names Y6 strain as Carnobacterium-like organism. PMID- 12548874 TI - [The construction of a recombined E. coli strain with stable and high production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)]. AB - Plasmid pJMC2 was constructed by cloning the parDE fragment of RK2 into pTZ18U PHB which harbored phaCAB from Alcaligenes eutrophus and was transferred into E. coli HMS174 and E. coli JM107 separately. It is very stable in its hosts cultured in medium without ampicillin. E. coli HMS 174(pTZ18U-PHB) and E. coli JM107(pTZ18U-PHB) produced P(3HB-co-3HB) in a low phosphate concentration medium(18 mmol/L). The proportion of 3-hydroxyvalerate(3HV) in the polymer was 5% 8%. A fed-batch culture of E. coli HMS174(pJMC2) was conduct in a 5 L automatically controlled fermentor, the final dry cell weight, P(3HB-co-3HV) content, and th 3HV proportion were 42.5 g/L, 70% and 4.9% respectively. PMID- 12548875 TI - [Cloning and sequence analysis of E0 gene of hog cholera virus lapinized Chinese strain and virulent shimen strain]. AB - According to the published nucleotide sequences of genome of hog cholera virus, one pair of specific primers were designed and synthesized. From the spleen of rabbits which were infected with HCLV and HCV Shimen strain infected pig blood, the two E0 genes were amplified by RT-PCR. The amplified fragments were cloned into pGEM-T vector and sequenced. Sequence analysis showed nucleotide sequence and deduced amino acid sequence homologies of the E0 gene between HCLV and Shimen strain were 95.0% and 94.3%. There was 13 amino acid substitutions between them. One N-glycosylation site was missing from E0 gene of the Shimen strain. The nucleotide sequence homologies of the Shimen strain with the ALD, GPE-, Brescia and Alfort strains were 97.4%, 96.5%, 92.2% and 86.5%, respectively, the deduced amino acid sequence homologies were 97.4%, 96.0%, 95.2% and 92.5%, respectively. The nucleotide sequence homologies of E0 gene of the HCLV strain with the ALD, GPE-, Brescia and Alfort strains were 95.6%, 94.9%, 91.3% and 85.5%, respectively, the deduced amino acid sequence homologies were 93.4%, 92.5%, 91.6% and 90.7%, respectively. E0 gene from HCV was shown to be similar to a family of fungal and plant ribonuclease. The catalytically important residues were 28 to 40 and 71 to 89 of HCV E0 gene. PMID- 12548876 TI - [The identification of BmNPV gp64 envelope gene]. AB - gp64 glycoprotein, an abundant and the only membrane glycoprotein encoded by the baculovirus genome, contains a signal peptide sequence at N-terminus, a transmembrane anchor at c-terminus and a ectodomain between them. The efficient signal sequences and the transmembrane anchor sequences of AcMNPV gp64 gene have been exploited in baculovirus expression system. Using 1.3 kb AcMNPV gp64 gene coding sequence as the probe, the nucleotide sequence of BmNPV gp64 gene was determined, and a 1530 nucleotide open reading frame flanked by an AT-rich sequence was identified that could encode a polypeptide with 509 amino acid residues. Computer analysis indicated BmNPV gp64 gene is 84.3% identical to AcMNPV at the level of nucleotide sequence and 94.7% identical at the level of predicted amino acid sequence. PMID- 12548877 TI - [Expression of recombinant human stem cell factor in insect cells]. AB - The cDNA of human stem cell factor(hSCF) containing signal sequence was cloned into the transfer vector pVL941 of AcNPV to construct a recombinant transfer vector pVL941-SCF. Sf9 cells were cotransfected with wild type viral DNA and pVL914-SCF to produce the recombinant virus AcNPV-SCF by homologous recombination in cell. Southern-hybridization analysis suggested that the recombinant viral DNA contained hSCF cDNA fragment. The Sf9 cells infected with the recombinant baculovirus AcNPV-SCF expressed biologically active rhSCF which was secreted into the cell culture. The synergistic activities of SCF in conjunction with human interleukin-3(hIL-3) was measured by MTT colorimetric method and TF-1 cell line. The expression level of Sf9 cells reached its highest at about 1970 units/ml in the 3rd day after the infection with AcNPV-SCF. Three SCF bands with molecular masses of 18 x 10(3), 20 x 10(3) and 22 x 10(3) were detected by immunoblotting. PMID- 12548878 TI - [Expression of Vitreoscilla hemoglobin gene in Streptomyces avermitilis]. AB - Two expression vectors, pWY101 and pWY102, were constructed by cloning Vitreoscilla hemoglobin gene(vhb) with its native oxygen-regulated promoter into E. coli-Streptomyces shuttle vector pIJ653. They were introduced into Streptomyces avermitilis, but Western blotting experiment failed to detect vhb gene expression. pHZ1252 is another shuttle vector for expressing VHb, in which vhb structural gene is controlled by a strong, thiostrepton-inducible Streptomyces promoter PtipA. pHZ1252 was transformed into S. avermitilis and expressed VHb which had biological activity after thiostrepton induction. pHZ1252 was structurally unstable in S. avermitilis, occurring deletion recombination. However, the rest part of pHZ1252 was stable in S. avermitilis and still contained vhb gene and PtipA. Plasmid pHZ1252 isolated from S. avermitilis was unable to transform E. coli, showing the loss of part of E. coli plasmid from pHZ1252. PMID- 12548879 TI - [Cloning and expression of the gene encoding maltoologosyl trehalose synthase from Sulfolobus shibatae in E. coli]. AB - 2.2 kb DNA fragment encoding a novel enzyme, maltooligosyl trehalose synthase (MTSase) was amplified from Sulfolobus shibatae by using PCR technique. The amplified 2.2 kb DNA fragment was inserted into an expression vector, pBV220, to yield the recombinant plasmid pSBGT1. MTSase gene in pBSGT1 was expressed in E. coli. The molecular weight of expressed MTSase detected by SDS-PAGE was about 74 kD, which is conformed with that deduced from nucleotide sequence. The expressed MTSase protein accounted for about 4.4% of the total cell protein. The MTSase from transformants containing pBSGT1 is capable of decreasing DE value, forming non-reducing or less-reducing saccharides when allowed to act on reducing partial starch hydrolysates. PMID- 12548880 TI - [Study on the production of beta-mannanase by Bacillus M50]. AB - Bacillus sp. M50 produced beta-mannanase efficiently in the medium composed of 4% konjac powder, 1.0% (NH4)2SO4 and 0.35% Na2CO3. Fermentation of beta-mannanase by Bacillus sp. M50 has been carried out in 100 L tank. The temperature, ventitation quantity and agitation speed were 30-32 degrees C, 1:0.75 vvm and 220 r/min respectively. the fermentation cycle was 28 h and the highest beta-mannanase activity was 330 u/mL. The optimal temperature and pH for beta-mannanase reaction were 50 degrees C and 6.0 respectively. It was stable at pH 4.0-7.0 and below 50 degrees C. The activity of the enzyme was strongly inhibited by Fe3+, Al3+, EDTA, Hg2+ and slightly stimulated by Ba2+, Mn2+. Treatment of ramie-fibers with crude enzyme preparations from Bacillus sp. M50 showed a good degradation for hemicellulose of ramie-fibers residues. PMID- 12548881 TI - [Purification and properties of beta-D-mannanase from Nocardioform actinomycetes]. AB - After cultivating in liquid medium for 3 days, an extracellular endo-1,4-beta-D mannanase (EC-3.2.1.78) from a Nocardioform actinomycetes strain NA3-540 was purified 137-fold to electrophoretic homogeneity by ammonium sulfate precipitation, 95% alcohol precipitation, CM-sephadex A-50, hydroxyaptite column chromatography, DEAE-cellulose anion-exchange column chromatography and Sephadex G-100 column chromatography. The enzyme had a molecular mass of 41 kDa(by SDS PAGE) and 40 kD (by gel filtration), a pI of 4.8 (PAGEIEF) and carbohydrates content of 3.5%. The enzyme was optimally active at pH 8 and 75 degrees C and showed stability at pH range of 6.5 to 12 at the temperature below 60 degrees C. The amino acid composition analysis of the enzyme proved that there were large amount of Gly, Asp, Ala and Glu. The enzyme was strongly inhibited by Hg2+, Cu2+, Pb2+, Fe3+, Co2+, Zn2+ and partly inhibited by Mn2+, Fe2+, Ag+ and slightly inhibited by Na+, K+, Li+, Mg2+, Ca2+, Ba2+. PMID- 12548882 TI - [Conditions of enzyme production and properties of alkaline lipase by Streptomyces Z94-2]. AB - An alkaline lipase producing strain streptomyces Z94-2 was selected from 152 lipase producer, The optimal medium composition for the producing lipase is (g/L): soybean meal 30, urea 10, dextrin 10, KH2PO4 0.5, NaCl 1, MgSO4.7H2O 0.5, AEO9 (nonionic surfactant) 1. The optimal conditions for lipase producing are initial pH 9.5-10.0, shaken at 26 degrees C, for 48 h, a maximal lipase activity of reached 596 (u/mL). The optimal activity at pH 9.8 and at 37 degrees C, respectively. The lipase was stable at pH 8.6-10.2 and below 60 degrees C, and the lipase was highly activated by 0.14 mol/L CaCl2. PMID- 12548883 TI - [Study on the physicochemical properties of Campylobacter jejuni enterotoxin]. AB - Precipitate of Campylobacter jejuni cytotonic enterotoxin(CE) performed in an 80% saturated solution of ammonium sulfateit indicated that there were some little molecular proteins except the 68 kD main band on sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE), whereas the eluate from GM1 ganglioside affinity column chromatography exhibited only one 68 kD band on SDS PAGE. The results suggest that CE mainly be consisted of 68 kD protein. The toxin is heat-labile, pH dependent and resistant to trypsin, It could be completely inactivated by heating at either 56 degrees C and 60 degrees C for 30 min or 100 degrees C for 15 min. The activity was maximum at pH 6.0 and was completely inactivate at pH 3.0 and pH 9.0, and rapidly reduced after storage over 3 d at 4 degrees C. The anti-LT serum could completely inhibited the activity of CE. PMID- 12548884 TI - [Screening of acrystalliferous mutants from Bacillus thuringiensis and their transformation properties]. AB - After elevating growth temperature of YBT-1463 and other 8 Bacillus thuringiensis parental strains to 42 degrees C, 9 acrystalliferous(Cry-) mutants were obtained, and then a series of partial plasmid-cured mutants were further obtained from 3 Cry- mutants BMB151, BMB154 and BMB160 by elevating temperature to 44 degrees C. One of which, BMB170, was further treated by elevating temperature to 46 degrees C and adding 0.05% SDS as the plasmid-curing agent. The resulting mutant, which was designated BMB171 and had no resident plasmid, was finally obtained. The studies on the transformation efficiency and stability of Cry- mutants showed that the transformation frequencies, as well as stability of several exogenous plasmids(pHT3101, pBMB1736, etc.) introduced into Cry- mutants by electroporation, were correlated with the numbers of resident plasmids bearing by the relevant recipients, the patterns of plasmid replicons and the sizes of those exogenous plasmids. Meanwhile, a much higher electro-transformation frequencies of Cry- mutants than the parental strain YBT-1463 was found, and the highest of the transformation frequency, in BMB 171, could reach up to 10(7) transformants/microgram DNA, and the stability of those exogenous plasmids in BMB171 were also higher than in other Cry- mutants and YBT-1463 strain. PMID- 12548885 TI - [Purification and identification of siderophore from Aeromonas hydrophila]. AB - Siderophore of Aeromonas hydrophila(Ah) J-1 was purified from culture supernatant by polyamide column chromatography, eluted by double-distilled water, ethyl acetate precipitated, lyophilized, then the white powder was collected, and demonstrated by CAS and Arnow assay. The siderophore was phenolate, composed of 2,3-dihydroxybenzoic acid (2,3-DHB), lysine, glycine and tryptophan by HPLC. Being cultured iniron-deficient medium, the purified siderophore could stimulated the growth of Ah N9a which siderophore production is poorly positive and reverse the inhibition growth of EDDA. It suggested that siderophore could promote growth of bacteria and might play an important role in bacteria infection. PMID- 12548886 TI - [Bacillus subtilis undergoes natural genetic transformation on agar plates]. AB - This paper conducted preliminary investigation on natural genetic transformation of Bacillus subtilis on agar plates. The results showed that, under same conditions, the natural transformation of the strain on agar plate was much more efficient than that of standard liquid method, and the transformation system could sustain higher DNase concentration. In addition, that the LB culture, which usually did not support the strain acquire competency, could undergo transformation as soon as it being spread on agar plate, suggested that the natural genetic transformation of Bacillus subtilis on solid plates may be different form the standard liquid transformation method. The cell-to-cell transformation between strains having different genetic makers could also be observed on agar plates. PMID- 12548887 TI - [The selection of fluoroacetate-resistant mutant from E. coli MMR204 and its influence on the expression of heterologous GL-7ACA acylase]. AB - In the cultivation of gene engineered strain of Escherichia coli on glucose medium, excretion and accumulation of acetic acid inhibit not only cell growth but also the the expression of heterologous protein. It is obvious that the desirable host strain maintaining acetate at a low level is one of the approaches to increase the production of recombinant protein. The present article deals with the selection of mutants of E. coli DP19, DP8, which grow on the medium containing pyruvate as the sole carbon source in the presence of 50 mmol/L fluoroacetic acid. It is shown that mutant DP19 is defective in its phosphotransacetylase(PTA) activity and accumulates less acetate in the medium, while DP8 is defective in acetate kinase (ACK) and accumulates similar level of acetate comparing with its parent. Using pta- mutant E. coli DP19 as host, the expression of GL-7ACA acylase gene on the recombinant plasmid pMR24 is improved, and the yield of enzyme activity in flask fermentation is about twice as much as its parent. PMID- 12548888 TI - [Research of toxin and plasmids of Saccharomyces cerevisiae]. AB - Killer toxin from Saccharomyces cerevisiae SK was isolated by ultrafiltration of culture supernatants and purified by poly(ethylene glycol). The toxin migrates as one single protein band on SDS-PAGE and its molecular weight is 15 kD. The SK toxin has the greatest lethal effect on the sensitive yeast strain in the lat-lag phase. Extraction and purification of killer heretity factor(dsRNA) from SK found that M-dsRNA plasmid and L-dsRNA plasmid have different molecular lengths being 1.7 kb and 4.0 kb. PMID- 12548889 TI - [Ribosomally synthesized antimicrobial peptide produced by eukaryotes]. PMID- 12548890 TI - [Molecular regulation of a factor in the differentiation and secondary metabolism of prokaryote]. PMID- 12548891 TI - [Studies on the biomedical sensor techniques for real-time and dynamic simultaneous detection and measurements of respiratory gases carbon dioxide and oxygen]. AB - Based on the studies of the direct detection of electro-inactive gas CO2 by amperometry and the mechanism of oxygen interference, a new modulated potential pulse coulometry/amperometry (MPPC/MPPA), which combines the fast modulated potential technique with the microelectrode method, has been advanced and established for the rapid determination of both CO2 and O2. The influence of modulated potential-time waveforms on the electrode activity, the accuracy and range of detection was investigated. Results were obtained which demonstrate that the novel biomedical sensor techniques can be used for the non-invasive, real time and dynamic simultaneous monitoring of respiratory gases CO2 and O2. PMID- 12548892 TI - [Bio-mechanical property changes of long bone in ovariectomized goats]. AB - This paper presents the biomechanical property changes of long bone in goats at 180 days after ovariectomy. 10 female goats at the age of 1.5 +/- years(average body wight 22 kg) were randomly divided into the control group and ovariectomized group. These goats were sacrificed at 180 days after ovariectomy and the femora, tibiae, metatarsus, humeri, radius and metacarpus were taken out bilaterally. The structural mechanical properties of these bones were measured with the three point bending method. The material mechanical properties of flexura, compress and tensile in the compact bone specimens taken from the two broken ends were also measured. The results showed that the bone density, bone thickness in broken ends, and the break load of three-point bending for the long bones in the OVX group were obviously lower than those in the control group (P < 0.05). And compared with the control group all the long bones in the OVX group, except the femur, had significantly lower ultimate bending strength, bending elastic modulus; ultimate flexural strength, flexural elastic modulus; ultimate compressing strength, and compressing elastic moduus. Some long bones in the OVX group had significantly decreased ultimate tensile strength (P < 0.05). These indicated that the mechanical properties of long bone in OVX group at 180 days after ovariectomy were reduced in varying degrees and were liable to fracture. This result is in accord with the clinical findings in osteoporosis resulting from post-menopausal estrogen fall and is also consistent with the osteoporotic model of ovariectomized rats. The authors conclude that the bone biomechanical properties are important parameters in assessing osteoporosis and the ovariectomized goat can possible be a good big animal model of osteoporosis. PMID- 12548893 TI - [Simulated experiment of hemodynamics changes in aorta during high energetic trauma through abdomen]. AB - In order to study the mechanism of remote effect of missile wound, we designed and carried out a simulated experiment. The change of hemodynamics in aorta was examined with a pressure transducer during gunshot and then calculated. The results revealed superhigh pressure, high speed of pressure rising, accelerated velocity of blood flow, substantial pressure difference and profuse blood flow in aorta during gunshot. PMID- 12548894 TI - [An algorithm for ECG data compression and its real-time realization with single chip computer]. AB - ECG data compression has been greatly improved since 1960 s. Although the error of compression can be controlled accurately, the reported algorithm at present can not be processed in real-time with a microcomputer. In this paper an algorithm for ECG data compression is expounded which can be realized in real time with a single chip computer. The algorithm has not only adjustable precision but also an average compression rate of 10.34, especially a pretty high compression ratio for frequent arrhythmic patients. PMID- 12548895 TI - [Application of time-frequency filter of wavelet to single-trail ERP extracting]. AB - A new approach to extracting single-trail event-related potential(ERP) in combination with the correlation analysis has been proposed. This approach, called time-frequency filter, is based on the concept that multiresolution wavelet transforms. At first, an investigation of the properties in time and frequency domain was carried out on the ERP. On the basis of the findings, the time-frequency filter of wavelet was designed and used for single-trail ERP extracting. The results and related discussion are presented in this paper. PMID- 12548896 TI - [Approach to recognition of ECG P waves based on approximating functions]. AB - A new method of recognizing ECG P waves based on approximating function is presented in this paper. The result of experiment demonstrates that the method based on the second order approximating functions to recognize P waves in useful and easy to do. The rate of correctness for recognizing P waves is higher than 99% according to MIT/BIH ECG Datebase. PMID- 12548897 TI - [The design and clinical application of an apparatus for monitoring neuromuscular transmission function during operation under general anaesthesia]. AB - This paper introduces a monitoring system of musle relaxant in action during the operation under general anaesthesia. The design of the monitor is based on measurement of acceleration. It consists of an acceleration transducer, a nerve stimulator and a microcomputer unit. The transducer can be fastened to the thumb, and the signal of the thumb movement in response to ulnar nerve stimulation is measured and registered in the computing unit. In clinical practice, it has been proved that this new apparatus can achieve reliable evaluation of nuromuscular transmission function and improve the quality of anesthesia in patients care during surgical operation. PMID- 12548898 TI - [Transient response in electric measurement of human auricular point]. AB - This is an experiment on the measuring E approximately t and R approximately t for human auricular point, which shows there is a transient response of electric potential E(t) and voltage drop U(t) when t < 2 tau and there is at intervals change with the time after 2 tau about R(t) and E(t). We have obtained a response function of the E(t) and U(t) by the circuit analysis. The parameter of the transient character is relaxation time tau, tau approximately RC. The mathematical model is in correspondence with the experimental result of the auricular point and TPM simulation. It indicates that the character changing with the time should be extracted after 2 tau. This experiment is of importance to understanding the electric character of the otopoint. PMID- 12548899 TI - [Comparing the diagnoses of pneumoconiosis made by computer and experts from X ray films]. AB - The results on the agreement between microcomputer and experts for the diagnosis of pneumoconiosis from X-ray films are presented in this paper. On a set of 200 films, the diagnoses made by the computer and by five experts were compared; From 100 films, the agreement rates were 83.5% for the normal/abnormal, 73.0% for the four categories of profusion and 70.5% for the four categories of pneumoconiosis. From 100 films, the rates of agreament between the computer and the basic-level consultation group were 86.0%, 82.0% and 63.0% respectively. From 300 films for screening pneumoconiosis, the rate of agreement between the computer and the experts was 89.3%. The results demonstrate that the automatic diagnosis by computer is closer to the diagnosis made by experts. PMID- 12548900 TI - [Design and laboratory manufacture of "SUU-Type I" extra-luminal flow hollow fibre membrane oxygenator with microporous polypropylene]. AB - The design and laboratory manufacture of "SUU-TYPE I" extra-luminal flow(ELF) hollow fibre membrane oxygenator are presented. The design is based on the O2 and CO2 transfer models in the cross-flow membrane oxygenator of Mockros and Pei. This paper also introduces the sketch of the membrane oxygenator, the structure of housing, the bundle of hollow fibres and the procedure of centrifugal potting. The self-made potting equipment and process of centrifugal potting are emphasized. The key know-how of centrifugal potting includes: (1) assuring the quality of the hollow fibre membrane material and the bundle; (2) calculating the quantity of potting material, polyurethane; (3) controlling reaction temperature; (4) removing gas from potting ends; (5) controlling rotation speed; (6) inserting the bundle in the house; (7) mastering the time of demoudling and cutting. PMID- 12548901 TI - [A micro transducer for dynamic multi-location pH monitoring in upper digestive tract]. AB - A new transducer catheter used for dynamic multi-location pH monitoring in upper digestive tract has been developed. On the catheter several antimony electrodes and an Ag-Agcl electrode are integreted. An animal experiment compared the results of gastric aspiration and dynamic recording and demonstrated the reliability of this transducer. Clinical application has shown that this technology contributes greatly to the research on digestive physiology and to the diagnosis of functional diseases such as non-cardiac chest pain. PMID- 12548902 TI - [A method of morphological observation on the endothelialization of bioprosthetic heart valve materials in vitro]. AB - In order to examine the endothelialization of yak pericardial materials in vitro, we used Wright staining and successfully observed the morphology, number and order of endothelial cells seeded on pericardial patches under light microscope. Compared with scanning electron microscopy, the method of light microscopy is simple, easy to do and may be carried out at any time. In laboratory, it brings much convenience in morphological observation on the endothelialization of biomaterials and therefore is worth recommendation. PMID- 12548903 TI - [Image and quantity analysis of prostaglandin in rats' blood plasma and Na(+) K(+)-ATPase in their cerebellum during the prevention of motion sickness by cinnarizine]. AB - To study the mechanism of cinnarizine in preventing motion sickness, TXB2, 6-Keto PGF1 alpha in rats' blood plasma and Na(+)-K(+)-ATPase activity in the endothelial cells of their cerebellar capillary were measured and analysed by a radioactive immunity analyser and a computer image system. The results showed that TXB2 and 6-Keto-PGF1 alpha in rats' blood plasma in the cinnarizine preventing group (CPG) decreased remarkably, compared with those in the motion sickness group(MSG) (p < 0.05). The activity of Na(+)-K(+)-ATPase in the endothelial cells of rats' cerebellar capillary in CPG was higher than that in MSG (p < 0.01). The authors suggest that the lower concentration of TXB2 and 6 Keto-PGF1 alpha in rats' blood plasma in CPG is closely related to cinnarizine which prevents Ca2+ from entering into the platelets and into the endothelial cells of blood vessels. The higher activity of Na(+)-K(+)-ATPase in the cerebellum may be caused by cinnarizene which dilates the blood vessels in the brain, increases the blood flow therein, and hinders Ca2+ from getting into the cerebellum cells. These change are believed to be the important mechanism of how cinnarizine prevents motion sickness. PMID- 12548904 TI - [Study on the method of adsorbing albumin for enwrapping human hard tissue substitute biomaterials]. AB - The aim of this study was to cover up the surface of human hard tissue substitute biomaterials with albumin. The cidex cross-link method was adopted to adsorb albumin so as to form a membrane for wrapping hydroxyapatite(HA), bio-glass ceramics(BGC), and hydroxyl poly-calcium sodium phosphate (HP). The results showed that this membrane of protein could enwrap the biomaterials so firmly that urea solution could not wash it off. PMID- 12548905 TI - [Implementing a relatively rigorous algorithm of dynamic EIT problem on the modified FEM model]. AB - We deduced a relatively rigorous algorithm of dynamic EIT problem-general inverse algorithm, and implemented it on the computer FEM model. Images which can show the changing of the resistivity(i.e. dynamic resistivity images) were gained. Also we modified the ordinary FEM model according to the characteristic of the EIT problem and implemented the general inverse algorithm on the modified model. The ill-posed condition of the forward matrix based on the modified model was fairly improved. It can be seen from the results that the general inverse algorithm with modified FEM model is an algorithm better than the back-projection algorithm in terms of object locating, image definition, and the ability of dealing with complex object and noise. PMID- 12548906 TI - [Integrating qualitative analysis with quantitative grey relation analysis in the comprehensive evaluation for solving medical problems]. AB - This paper integrates qualitativ analysis with quantitative grey relation analysis in the comprehensive evaluation for solving medical problems, especially diagnostic problems, and good results are obtained. In it, some new medical concepts such as diagnostic value, diagnostic grey degree and information cover rate are defined. The given example demonstrates that when used in clinical diagnosis, this diagnostic model makes a feature of high diagnostic coincidence rate as well as low misdiagnosis rate and it overcomes the common disability of general medical diagnostic model in quantitative analysis. PMID- 12548907 TI - [The three-dimensional analysis of mandibular overdenture supported by implants]. AB - In recent decade, the treatment in which two or four implants are placed in an edentulous patient to support an overdenture has been more effective and safe, and the retention and stability of overdenture have been improved by the use of attachments fabricated on implant abutments. This study aimed to investigate the supporting tissue stress distributions of implant-overdenture with a 3-D finite element model built by CT scan and CAD technology when various attachments and loads were used respectively. The results demonstrated that the establishment of the three-dimensional finite element model with CT scan and CAD technique had saved time and effort, and it did not damage the experiment model. The tissue structure was quite distinct in imaging. In terms of the stress distribution of bone tissue in the overdenture supported by implant, there was a difference between bar attachment and stud attachment. However, such a difference was minor as compared with the difference caused by the bite force direction. The stress value of cortical bone around abutment with 20 degree oblique force was 2.2-3 times that with vertical force. Therefore, to reduce high stress peaks, attention must be paid to the direction of the bite force, as this variable is much more important than the design of superstructure. PMID- 12548908 TI - [The changes in quantities of elements around cementum-dentin junction in human natural tooth--electron probe microanalysis]. AB - To study the changes of inorganic and organic substance around cementum-dentin junction in the tissues of human natural tooth, the method of electron probe microanalysis was used in this research project. Determinations of weight percentage (wt%) were made for Ca, P and S elements around cementum-dentin junction. The results showed that the changes in quantities of Ca, P and S elements around cementum-dentin junction were not significant(P > 0.1). These suggest that the quantities of inorganic and organic substance have not obviously varied with the anatomic structure of natural tooth. PMID- 12548909 TI - [Measurement of interfragmentary micromovement induced by mechanical vibration with accelerometers in vitro]. AB - The interfragmentary micromovement induced by mechanical vibration was measured in vitro in seven preserved adult cadaveric femurs by using accelerometers. Accuracy analysis of this method indicated that it can detect relative interfragmentary micromovement as small as 0.1 mm or below under certain range of frequency. Also, the results have shown that the mechanical vibration can induce interfragmentary micromovement under intramedullary nailing, but it can not do so under plate fixation. PMID- 12548910 TI - [Chitosan pin fixation of rabbit proximal tibia osteotomy]. AB - Degradatable chitosan pin, 4 millimeters in diameter and 30 to 40 millimeters in length, was studied radiographically and histologically (oxytetracycline labeling) at 3, 6, 12, 24 and 48 weeks after the implantation for fixation of a transverse proximal tibia osteotomy in fourty rabbits. The results showed that new bone formation and oxytetracycline uptake were increased 3-6 weeks after operation and the fracture was completely healed 12 weeks after operation, but in two rabbits the chitosan pin broke, in 3 weeks. Histologically, there was a massive inflammation (non-specific foreign body reaction) 3-12 weeks after implantation, which gradually subsided 24 weeks aften operation. Chitosan pin was slowly degradated in bone and was not replaced by normal tissue 48 weeks after operation. The questions to be addressed in future include how to increase the mechanical strength of chitosan pin and how to improve the characteristics of its surface. PMID- 12548911 TI - [Biodegradation and biocompatibility of a chitosan film]. AB - The biodegradability and biocompatibility of a chitosan film were investigated in mice. The results showed that chitosan films had a mild inflammatory reaction in the early days of grafting, and after 16 weeks the inflammation basically subsided. Chitosan films were easily biodegraded. Chitosan is a novel natural absorbable medical film material and has a good developmental prospect. PMID- 12548912 TI - [The safety interlock designed for JX-100 X-knife system]. AB - This paper describes the design of a safety interlock used by X-knife system for preventing the linear accelerator gantry colliding with the treatment couch and the patient when the accelerator is rotating and irradiating X-rays. How to install and adjust the unit is also addressed in this paper. The unit has been successfully used in JX-100 X-knife system developed by the First Military Medical University. PMID- 12548913 TI - [A new target position simulating system of X-knife]. AB - This acticle describes a new target position simulator system of X-knife. It adopts a new method of simulating with target ball to ball hole as a substitute for the traditional way of simulating with target pointer to pointer. It has the advantages of precise localization, convenient operation and dependable repeatability. Clinical application has demonstrated that it is an ideal target position simulating wystem of X-knife. PMID- 12548914 TI - [Development and clinical significance of SEA-1 emergency micro-ventilator]. AB - To improve the work of emergency care and early cardio-pulmonary-brin resuscitation in our county, which is characteristic of abruptness and contingency, a new type of ventilator is highly required. It is hoped that such a ventilafor has a simple safe valve and does not need electric current from the mains. Also, the ventilator should be small in size, light in weight, simple in operation, easy to carry, swift to assembly, and reliable in ventilation. With the principle of pneumatic logic elements and the design of the integration of pneumatic circuits, we have successfully developed the Emergency Micro ventilator, which accords with the above requirements. It has been confirmed that the ventilator is very effectual and reliable in ventilation support for the patients(n = 55) under general anesthesia without any case of hypoxemia, hypercapnia, hypotension, arrhythmia and so on, and the mechanical performances are stable. PMID- 12548915 TI - [The method and development of computer-assisted surgery]. AB - The methodology and the state of the art of Computer-Assisted Surgery (CAS) are introduced in this paper. Computer-assisted surgery is a new high technology which uses computer science, biomedical engineering, mechanism, mathematics, surgery, and so on. Its objective is to help surgeons use multimodal data, such as CT, MRI, DSA, PET, et al. in a rational and quantitative way in order to plan and perform medical intervention. Stereotactic localization method and registration are two cruxes in computer-assisted surgery. There are several methods for localization and registration. In recent ten years, computer-assisted surgery has been a cynosure of scientists. Some computer-assisted surgery systems have been used in clinical practice. PMID- 12548917 TI - [Fully play the role of science and engineering institutes in medical modernization]. PMID- 12548916 TI - [Research on fully automated gene analysis system]. PMID- 12548918 TI - [Microbial succession on lignite along with weathering]. AB - Six different weathered lignite samples were examined by scanning electron microscopy. Few microorganisms were observed on lignit just excavated and only spores and short hyphae were observed on lignite samples excavated 5 months, 1 year and 4 years before sampling. When lignite samples were moistened with distilled water and incubated for 10 days, actinomycetes proliferated significantly on lignite samples that were just excavated or excavated 5 months before sampling. The growth of bacteria was observed on lignite samples excavated 1 year before sampling. Fungi increased in length and in number on lignite samples excavated 4 years before sampling. These findings of microbial succession on lignite samples along with weathering were consistent with results of the plate count method; actinomycetes are the first colonizers, then bacteria and fungi are the last degrader. The dominant microorganisms were Actinosynnema sp., Streptomyces sp. and Nocardia sp. among actinomycetes, two Arthrobacter sp. among bacteria and two Aspergillus sp. among fungi. PMID- 12548919 TI - [Cloning and sequencing of the Chinese bovine prion protein (PrPC) gene]. AB - The total DNA was isolated from lymphocyte of peripheral blood of Chinese cattle. The PrPC gene was amplified by polymerase chain reaction, using two primers. It was then cloned into pGEM-T Easy Vector. The result of DNA sequencing was indicated that the 795 bp amplified fragment contains the entire PrPC coding sequence, which has no intron and is same as the published gene sequence. PMID- 12548920 TI - [Ultrastructural differences of RMV and TMV infected Nicotiana tabacum mesophyll cells for distinguishing virus strains]. AB - Nine strains of ribgrass mosaic virus (RMV) and tobacco mosaic virus(TMV) from cruciferous and solanaceous plants in different geographic regions of China were used to inoculate 4-leave-old plants of Nicotiana tobacum for observation of ultrastructually pathological changes due to infection of different strains. The samples of the mesophyll cells of the plants 15 d after inoculation were prepared in a routine and examined using transmission electronic microscopy. Characteristic virion arrangement, inclusion forms, and cell structural aberrations were found different among the various strains. The virions were accumulated in cluster, arranged in ring scattered, irregularly congregated with membrane, and aggregated in angle layer or parallel layer in cytoplasm and vacuoles whereas they were not found in organelles including nuclei, chloroplasts, and mitochondria. The inclusions with bacilliform, club-shaped or granulous X-tubule varied in number and form among the viral strains investigated. The abnormality of cell wall including thickening, collapsing and distortions were also observed among the cells infected by the strains. Other distinguishable changes attributed to the strains included the number, size, shape, and clumping pattern of chloroplasts, and of starch grains and osmiphilic globules chroloplasts of mesophyll cells infected only by two of the 9 strains examined. Additionally, the increased numbers of mitochondria and endoplasmic reticulum were observed in the cells infected by some of the strains. The results indicate that the ultrastructural changes due to viral infection may be useful for distinguishing various RMV and TMV strains. PMID- 12548921 TI - [Structure and function of saw1--a gene related to differentiation of Streptomyces ansochromogenes]. AB - The DNA sequence of double strands of saw1 gene was determined by the method of chaintermination inhibitors. The result indicated that the 1500 bp of DNA fragment displayed a complete open reading frame (ORF), the encoding regions were located in 419 to 1252 bp positions, saw1 product has 89% identity to amino acids of sigma whiG of Streptomyces coelicolor. When recombinant plasmid containing a 1500 bp saw1 DNA fragment insert in plasmid pIJ702 was introduced into Streptomyces coelicolor C71, the sporulation deficient mutant C71 could form spore and produce grey pigment. The gene function was studied by using the method of gene disruption and the result showed that the saw1 played an important role in developmental changes from aerial hypha to spore formation of Streptomyces ansochromogenes. PMID- 12548922 TI - [Cloning and expression in E. coli of 2,5-DKG reductase I from Corynebacterium]. AB - Two different 2,5-DKG reductases was purified from Corynebacterium sp. SCB3058, then its genomic DNA was used as template, a segment containing 2,5-DKG reductase I was amplified by PCR and cloned into pGEM3zf(+) to obtain the recombinant plasmid pGEM813. The sequence analysis showed that the cloned segment was 1107 bp in length, contained a single open reading frame of 834 nucleotides, which encoded a 34 kD protein consisted of 278 amino acids. After the primary control sequence was deleted, the expression vector pBL4 was constructed by plasmid pBL. With induction of temperature, a 34 kD protein was specifically expressed in E. coli DH5 alpha in high level. The expressed recombinant protein accounted for 20% of the total cell protein and had high specific enzyme activity. In spite of 30 degrees C, 37 degrees C or 42 degrees C induction, the specific activity of enzyme was almost the same level. These results indicate that most of the recombinant protein induced at 42 degrees C was existed with inclusion bodies. This work was the bases of constructing a recombinant Erwinia which can produce 2 KLG directly from D-glucose by one-step fermentation. PMID- 12548923 TI - [Studies on characterization of glutamine synthetase from Streptomyces lincolnensis]. AB - Glutamine synthetase from Streptomyces lincolnensis was purified to electrophoretic homogeneity. The effects of pH, temperature and metal ions on the purified enzyme were studied, and the substrate specificity of glutamine synthetase were reported. Under the standard biosynthetic assay system, the substrate saturation curves of glutamine synthetase from Streptomyces lincolnensis for ammonia did not agree with the Michaelis-Menten relationship, with either Mn2+ or Mg2+ as divalent cation, and Hill constants not equal to 1. The K(m) values of GS for Glu and ATP were different with different divalent cation. PMID- 12548924 TI - [Nuclease P1 production by immobilized Penicillium citrinum cells]. AB - Mycelia of Penicillium citrinum were adsorbed and immobilized efficiently within porous polyurethane. Nuclease P1 produced by the immobilized cells were studied in shaking flasks. The suitable glucose and peptone contents in medium were 10 g/L and 1 g/L, respectively. And the shaking speed was 180-200 r/min. After 48 h fermentation, the nuclease P1 activity reached 513.3 U/ml, the productivity was 3.6 times higher than that of the free cells. The production cost is obviously reduced. In repeated batch fermentation, the immobilized cells kept the capacity after 28 batches for 56 days with an average nuclease P1 activity of 507.4 U/ml. PMID- 12548925 TI - [Study of the physiological characteristics of a strain which can change the color of red lead on Dunhuang mural]. AB - A strain which was isolated from Dunhuang Murual was investigated. It belongs to Flavobacterium. In media added in Pb3O4, colonies show brown and black. After identified pigments, it showed that the brown was caused by the oxidation of Pb3 O4 into PbO2 by the strain. The optimal oxidative condition is at pH 9.8, 37 degrees C and dark environment. The genes for oxidation is on plasmid and the oxidation was inhibited under pure O2 and N2 conditions, too 5 x 10(-3) mol/ml NaN3 is enough. The strain was able to absorb lead initiative. The transmission electron microscope showed that lead locate in protoplasts. PMID- 12548926 TI - [Reliability of extracellular protease and lipase activities of Beauveria bassiana isolates used as their virulence indices]. AB - The extracellular protease and lipase activities of 17 Beauveria bassiana isolates from different hosts and countries were evaluated for the reliability for the indices of their virulence to the migratory grasshopper, Melanoplus sanguinipes. Virulence assay of each isolate included about 30 10-d-old grasshoppers receiving topical inoculation with the suspension of 10(7) conidia/ml. In the assays of the enzymes, N-succinyl-Ala-Ala-Pro-Phe-p nitroanilide and p-nitrophenyl palmitate were used as a substrate to measure the activities of protease (3 replicates) and lipase (4 replicates) in the filtrates of gelatin-based and sunflower oil-based liquid cultures of each isolate, respectively. Varying among the isolates assayed, the estimates of LT50's, protease units (PU), and lipase units (LU) were 5.27-16.89 d, 0.47-3.37 x 10(-2) mumol.ml-1.min-1, and 0.00-56.75 mumol.ml-1.h-1, respectively. Regression analysis revealed that PU was significantly (P < 0.01) correlated to the daily cumulative mortality of M. sanguinipes 5-17 d after inoculation and the LT50's whereas LU had little correlation to either the mortalities or the LT50's (P > 0.10). Based on the determination coefficients (r2) from the regression, PU alone interpreted at most 67% of the variation in the mortality 7d after inoculation but less than 50% in most of the days considered and only 38% in LT50's. Thus, the author suggested that PU could be used as virulence index only for early stage selection of candidate isolates in large quantity and could not entirely replace conventional virulence assay method that remains most reliable. PMID- 12548927 TI - [Isolation, detection toxicity and structure of toxin from Beauveria bassiana]. AB - In this experiment, the crude toxin from the metabolite of Beauveria bassiana was isolated and its toxicity was detected. The result shows that indicates toxin has toxicity on larva of Aedesalbopictus obvious. Though it with low toxicity to larva of Heliothis zea when swallowed, but has high toxicity when injected into the blood. For bacteria it only inhibites G+. When it applied to cultured cells of Spodoptera frugiperda sf-21, the regression equation of toxicity is y = 2.03 + 2.39x. The half lethal concentration (LC50) is 0.01751%. 95% believable limit is 0.01517-0.0202%. There are three substances which have similar atomic group when analysed by HPLC and infrared spectrum. PMID- 12548928 TI - [Study on HCMV replication in gene transtecfed cells]. AB - Five gene transfected cells (GTC: A5, B3, D3, G8 and H3) were generated by DNA (from HEP-2) transfer (into HEL). Chromosome number of the transfectants fell between that of HEL and the combined chromosome number of the two cell lines. D3 showed more positive fluorescence cells than A5, B3, G8 and H3 by 4d after infection of HCMV. As HCMV infection dose was increased from 0.1 to 100 PFU/ml, significant increase of viral production was observed. The similar susceptibility to HCMV was shown in different passages of D3. The immortal D3 may provide a tool to analyze host cell factors controlling the transcription and replication of HCMV. PMID- 12548929 TI - [Isolation and identification of N2-fixing strains of Bacillus in rice rhizosphere of the Yangtze River Valley]. AB - Rice rhizosphere soil samples were colected from 10 sites of 7 provinces in the Yangzi River Valley, and from the soil samples 16 endospore-forming strains with ARA (Acetylene Reduction Activity) were isolated, the nitrogen fixing ability was tested by the method of 15N tracer and the atom 15N% excess are ranged from 0.0297% to 0.4714%. The strains were identified as Bacillus licheniformis, B. subtilis, B. azotoformans, B. cereus, B. pumilus, B. brevis, B. megaterium, B. firmu. PMID- 12548930 TI - [Selection of ningnanmycin overproduction strains by resistance of self-secondary metabolites]. AB - The S. noursei var. xichangensis FR-5 protoplasts was treated by UV, NTG in sequence, and the mutated strains were selected by use of self-secondary metabolites, the microorganisms were isolated by fermentation after agarblock rational method. The seven strains of microorganisms which were one fold production higher than that of original strain were obtained from 122 mutants. The method should be a simple, high speed selection model of breed Ningnanmycin overproduction strains of microorganisms. PMID- 12548931 TI - [Taxonomy of a new species of haloalkalophilic archaeon]. AB - A strain of Haloalkalophilic Archaeon Y21 was isolated from soda lake in Inner Mongolia Autonomous Region. The isolate is rod shaped, motile and red clone. On the basis of morphology, physiology and biochemistry, polar lipid composition, phylogentic analysis and DNA-DNA hybridization, we found that strain Y21 is new species of the genus Natrilba. The name Natrilba wudunaoensis sp. nov. is proposed. PMID- 12548932 TI - [Expression, purification and characterization of fruA, a transcription factor in Myxococcus xanthus]. AB - FruA is a transcription factor essential for the development of Myxocoxccus xanthus. Gene encoding fruA with a poly-histidine tag was expressed in E. coli and simply purified by chromatography on nickel column. Data from gel retardation assay suggest that FruA regulates transcription of target genes in collaboration with other factors. PMID- 12548933 TI - [Cloning and expression of a resistant Culex gene in Escherichia coli]. AB - Recombinant plasmid pRLB1 was constructed from detoxifying gene B1 of pesticide resistant Culex pipiens quinquefasciatus and from plasmid pRL439 contained the strong promoter PpsbA. The positive clone was identified by digestion and Southern analysis. Expression of recombinant plasmid containing esterase gene was detected. An engineered bacterium possessing high enzyme activity was obtained and immobilized. It can effectively degrade the specific substrate alpha-naphthyl acetate (alpha-NA) and beta-naphthyl of esterase enzyme. Assays showed that pesticide acetofenate (7504 an organic choride pesticide) was degraded by the immobilized cells within one hour. PMID- 12548934 TI - [Expression and activity analysis of the human glutathione-S-transferase in Lactococcus lactis]. AB - The glutathione-S-transferase A1 cDNA was amplified from human liver total RNAs by RT-PCR and was cloned into a Escherichia coli expression vector pET23b, then the recombinant plasmid pET23bhgst was introduced into E. coli BL21 (DE3) and induced by IPTG, the high-level expression of hGSTA1 appeared in the E. coli cells. The cDNA encoding hGSTA1 was subcloned into pMG36e, a lactococcal expression vector, and introduced into Lactococcus lactis MG1363 by electroporation. In the positive transformants, the hGSTA1 was expressed as a fusion protein which was verified by SDS-PAGE and Western blot. The hGSTA1 from both E. coli and L. lactis was purified by affinity chromatography on glutathione agarose and all showed enzymatic activity. The potential application of the recombinant Lactococcus lactis in functional food was also discussed. PMID- 12548936 TI - [Studies on detection methods of two DNA probes in Mycobacterium tuberculosis]. AB - Two different biotinylated DNA probes which are highly specific to M. tuberculosis(MT) were made and studied. One probe is a 20 bp oligonucleotide labeled with biotin at 5' end, the other is a long DNA probe produced by PCR amplification procedure allowed for the incorporation of biotin labeled UTP. The two probes were hybridized with MT genome DNA and a 317 bp PCR product amplified from IS6110 sequence of MT, and then detected by alkaline phosphatase conjugates through colorimetric reaction. The detection sensitivity and specificity of the two probes were comparatively studied. The hybridization condition including concentration of probe, temperature of hybridization and washing filter thereafter were also investigated preliminary. The detection limit of the oligonucleotide probe and the 188 bp PCR probe were 100 ng and 6 ng of DNA respectively in detection of M. T genome, and 400 pg and 50 pg of DNA respectively in detection of PCR products of MT. The two probes can be only hybridized to MT and BCG, but not with other 24 mycobacterium or non mycobacterium tested. The optimal hybridization temperature and washing filter temperature of oligonucleotide were 42 degrees C and 60 degrees C respectively; and that of 188 bp probe, 68 degrees C and 60 degrees C-68 degrees C. Generally the specificity of two probes were all high, but the sensitivity of 188 bp DNA probe was 7-16 times that of the oligonucleotide probe. The higher sensitivity, lower hybridization background and faster revelation of the 188 bp DNA probe made it a better choice in detection of MT. PMID- 12548935 TI - [Expression and insecticidal characteristics of cry3A in Lepidoptera-specific Bacillus thuringiensis]. AB - The cry3A gene coding Coleoptera-specific protein was transformed into a wild strain YBT803-1 by electroporation, A transformant BMBY-001 was obtained. SDS PAGE analysis demonstrated that cry3A could express 64 kD protein in BMBY-001. However, the expression of cry1Ab and cry1Ac were effected. Bioassay showed that BMBY-001 was not only highly toxic to Phyllodecta vulgatissima larva(LC50 0.413 microL/mL) but also toxic to Plutella xylostella(LC50 3.319 microL/mL). PMID- 12548937 TI - [Boot colonization of wheat by lux-AB genes marked Pseudomonas fluorescens Xl6L2]. AB - Colonization density of Pseudomonas fluorescens Xl6L2 marked with luxAB genes in wheat rhizosphere in asepsis rhizobox-Calcareous chao soil microcosms reached the maximum(4.60 log cfu.g-1 root) 3 days after seeds coated with Pf.Xl6L2 sown, then declined to a relatively stable lower level(2.45 log cfu.g-1 root) in 0-2 cm root segment. Dynamics of colonization of Pf.Xl6L2 in wheat rhizosphere under field conditions was the same as in microcosms, Pf.Xl6L2 could move to the place of 10 cm of root from seeds under field conditions, distance of horizontal movement of Pf.Xl6L2 in field soil was not over 40 cm during 125 days of plant growth. PMID- 12548938 TI - [Construction of a novel Bm NPV Bac to Bac system]. AB - A Bi-Shuttle vector Bm-Bacmid was constructed by co-infecting Bm N cells with wild type genomic DNA from BmNPV and Ac-Bacmid DNA. It could not only replicate in E. coli cells as a large plasmid and but also remain infectious when induced into Bm N or Sf9 cells. Recombinant virus rBmHBe was obtained after transposition of a donor plasmid carrying Hepatitis Be antigen gene (HBeAg) into att Tn7, and was demonstrated by Southern blotting. SDS-PAGE analysis showed that HBeAg gene were highly expressed in Bm N cells. By ELISA testing, the highest antigenecity titer of HBeAg protein in cell cultural medium was up to a dilution of 1:32,000. Although HBeAg protein also presented in the Bm N cells the titer was only 1:2000. The HBcAg protein was much fewer than HBeAg (< 1:160) whatever in culture medium and in cells. The results showed that Bm N cells was able to recognize the signal peptide sequence and cut it correctly for HBeAg protein's excreting production. PMID- 12548939 TI - [Characterization and transformation of anti-FOA strain of Aspergillus awamori]. AB - A suspension of freshly harvested spores of Aspergillus awamori SG1 strain was exposed to UV light. The irradiated spores with survival rate of 10%-15% were plated directly on minimal medium (MM) agar plates containing 10 mmol/L uridine and 5-fluoro-orotic acid (FOA 1 mg/mL). Five stable anti-FOA colonies with the reversed rate below 10(-9) were obtained. All anti-FOA strains, can grow on uridine or uricil-containing minimal medium agar plate but not on orotic acid containing MM plate, indicating that the mutation happened in the URA3 gene. Northern blotting analysis showed that the mutation of URA3 gene could not be transcribed and it was confirmed by RT-PCR. With modified PEG-mediated protoplast transformation method the wild URA3 gene could complement the ura3 mutant strain SA5 to wild type. Southern blotting of the transformant indicated the wild RUA3 gene replaced the mutated ura3 gene. PMID- 12548940 TI - [Production of monoclonal antibodies to broad bean wilt virus and application in virus detection]. AB - Six hybridoma cell lines secretion monoclonal antibodies(MAbs) against broad bean wilt virus(BBWV) were produced by fusing mouse myeloma cells(SP 2/0) with spleen cells from BAL B/c immunized by the BBWV particles. The hybridoma cell lines secreted MAbs stably after cultured in vitro for 3 months or stored in liquid nitrogen and then revived for several times. The titres of ascitic fluids of six MAbs ranged from 1:256,000 to 1:640,000 when measured by indirect ELISA. In agarose gel immunodiffusion test, it showed that the six MAbs represented the same isotype of murine antibodies, IgG1. Six MAbs could detect 4 tested BBWV isolates, but didn't crossreact with other 5 plant viruses. The result of Western blot showed that all the six MAbs can react with the 44.7 kD large coat protein subunit of BBWV. This is the first report of production of MAbs against BBWV. PMID- 12548941 TI - [Study on the plasmid instability of Bdellovibrio BDG-9]. AB - A new method system was established in this paper to study the plasmid in stability of Bdellovibrio BDG-9. Using this system, it was found that when BDG-9 was cultured singly on the SMB plate, plasmid pST I was unstable although pST I still replicated and distributed to progeny cell normally. The results showed that the pST I copy number in single cell of BDG-9 decreased gradually to zero with the propagation of BDG-9. Additionally, plasmid pST I was very important for the growth of BDG-9, and with the lacking of pST I s, the growth and propagation of BDG-9 ceased. PMID- 12548942 TI - [The key enzymes of metabolisms of glycerol in Candida glycerolgenesis]. AB - The specific enzymes for glycerol formation and dissimilation in Candida glycerolgenesis were studied. C. glycerolgenesis hardly assimilated glycerol without glucose and a little amount of glucose remarkably stimulated this dissimilation pathway and the activity and expression of mitochondrial glycerol-3 phosphate dehydrogenase was induced strongly by glycerol-3-phosphate and repressed by glucose metabolism. During glycerol production fermented by C. glycerolgenesis, the activity of cytoplasmic glycerol 3-phosphate dehydrogenase maintained at a higher level and existed two peaks of activity, which appeared at 36 h and 60 h, respectively. The activity level of the first activity peak determines glycerol productivity and becomes a key enzyme of glycerol formation. The activity of glycerol 3-phosphatase maintained at a higher level just between 18 h and 48 h and appeared on peak activity at 36 h. In that period, the activity of glycerol 3-phosphatase was up to more than two times of cytoplasmic glycerol 3 phosphate dehydrogenase and that was inosculated with the phase of rapid accumulation of glycerol during glycerol production by C. glycerolgenesis. In the phase of slowly accumulation of glycerol between 48 h and 72 h, the activity of glycerol 3-phosphatase decreased below that of cytoplasmic glycerol 3-phosphate dehydrogenase. So, cytoplasmic glycerol 3-phosphate dehydrogenase is a key enzyme of glycerol formation in the phase of rapid accumulation of glycerol and glycerol 3-phosphatase becomes a more effective factor after glycerol fermentation reached into slowly accumulation phase. In a conclusion, it is a fundamental for C. glycerolgenesis over-producing high level of glycerol that its cytoplasmic glycerol 3-phosphate dehydrogenase maintains at high activity and the activity of glycerol 3-phosphatase is much higher than that of cytoplasmic glycerol 3 phosphate dehydrogenase. PMID- 12548943 TI - [Production and some properties of a thermophilic protease from Bacillus stearothermophilus WF146]. AB - The factors affecting Bacillus stearothermophilus WF146 for thermophilic protease producing have been investigated, more than 600 units of enzyme in 1 mL of fermented culture could be achieved under suitable condition. The protease had a molecular weight around 34 kD estimated by SDS-PAGE, and functioned optimally at pH 8.0 and 80 degrees C, respectively. In addition, the enzyme exhibited high temperature tolerance and was stable at a wide range of pH, and Ca2+ played a key role for the stability of the enzyme. While the protease activity of the enzyme was strongly inhibited by PMSF, DFP and IAA, and was not affected by DTT. PMID- 12548944 TI - [Enzymatic synthesis of saccharide and saccharide alcohol fatty acid esters]. AB - Conditions for esterification in tert-butanol using fatty acid as acy1 donor and saccharides or saccharide alcohols as acy1 acceptors, immobilized lipase from Candida sp. 1619 as biocatalyst were investigated. Optimal temperature and pH for esterication were 40 degrees C-45 degrees C and 5.5-7.5, respectively. Among the 13 acy1 donors compared, the best ones are the linoleic acid and oleic acid. The similar conversion degree was obtained when using saturated fatty acids with carbon chain length from C8 to C22. Among the 23 saccharides and saccharide alcohols compared, D-fructose, D-Xylose, trehalose, D-sorbitol, xylitol, mannitol isopropyidoneglucofruranose and alpha-methylglucopyranoside showed much higher esterification degree than other acy1 acceptors. In addition, the reaction degree of saccharide alcohols were substantially higher than that of the corresponding saccharides. The conversion was enhanced greatly when the molar ratio of acy1 donor to acy1 acceptor was higher than 2 to 1. In the case of using sorbitol and loeic acid as substrates, the reaction was carried out in 100 ml conical flask and the reaction system consisted of 3.0 mmol oleic acid(0.85 g), 0.2 mmol sorbitol(0.036 g), 3 mL tert-butanol, 30 mg immobilized lipase(600 u), shaken at 40 degrees C. After reaction for 48 h, more than 90% of sorbitol was esterified based on equal molar of substrates. The products were identified TLC as monoester and diester. PMID- 12548945 TI - [Continuous ethanol fermentation using tapioca starch by immobilized yeast cell without carrier]. AB - A widespread interest has been noted in continuous power ethanol fermentation utilizing systems with immobilized yeast cell through self-flocculating. We studied continuous ethanol fermentation by a naturally flocculent strain of Schizosaccharomyces pombe and the performance of two-stage continuous ethanol fermentation system using suspended-bioreactors(total effective volume 1.5 L) has been established: the high biomass levels achieved allow efficient ethanol conversion(72.7 g/L, average), residual glucose(3.74 g/L) and high productivity(9.7 g/L.h) for 150 g/L glucose concentration in feed(tapioca syrup) with average 90% total conversion of substrate. The system was run steadily during a month research. PMID- 12548946 TI - [Studies on the trehalose-producing by Colloides CB39]. AB - A bacterium which had the ability of producing sugar produced sugar at low temperature (18 degrees C) was selected from the water of Tianchi in Changbai mountain. The sugar was identified to be trehalose by methods of thin-layer chromatography, reaction of producing osazyone, capillary electrophoresis and infrared spectrum. It was also been found that the trehalose produced by this bacterium (identified to be a new species of Colloides Sp. CB39) was exocrine. At 18 degrees C, its trehalose content in culture solution was 256 mg/g dry weight of the strain. This characteristic is different from that of other strains, which had been proved to have the ability of producing trehalose. then the strain CB39 was induced by ultraviolet in order to abstain mutants which can produce trehalose in high level at usual temperature. The mutant strain 5 that could produce trehalose in high level at usual temperature (25 degrees C) was selected from all the mutant strains. The value of its productivity of trehalose is 416.7 mg/g dry weight of the strain. (At the same temperature, trehalose productivity of mutant strain 5 was eight times of that of the wild type strain CB39). PMID- 12548948 TI - [Effect of sugars and pH on the fermentation of decane 1,10-dicarboxylic acid]. AB - Some sugars, such as lactose, maltose, raffinose, glucose etc. can increase the purity of DC12 compared with sucrose. The value of pH can affect the process of producing DC12. Controlling the pH value between 7.5-8.0 can highly improve the production of DC12. Candida tropicalis can symport the DC12 with H+ to the outside of the cell. PMID- 12548947 TI - [Interaction between biofilm formed by Pseudomonas aeruginosa and antibacterial agents]. AB - We hypothesized that bacterial biofilm formation could be an important factor that make some infection intractable. Interaction of P. aeruginosa biofilm with antibactial agents was examined in vitro. METHODS: P. aeruginosa incubated for 6 days in a saline-teflon system formed biofilm on the surfaces of the teflon piece. The biofilm were characteried by dense colonization of bacteria and thick membranous structure that covered the colonies, it was observed with a scanning electron microscopy. CPFX, CAM, ROX and Herba Andrographitis were added to the biofilm bacteria. RESULTS: The rate of survial of the floating bacteria decreased to 0.02% after 4 hours of contact with 2MIC of Ciprofloxacin (CPFX), but that of biofilms was 41% When 10 micrograms/mL of Clarithromycin (CAM per mL was combined with CPFX, the survial rate for the biofilm bacteria decreased to 0.2%. When 12 micrograms/mL of Roxithromycin (ROX) was combined with CPFX, the survial rate for the biofilm bacteria decreased to 0.5%. When 0.05 g of Burm. f. per mL was combined with CPFX the survial rate for the biofilm bacteria decreased to 2.7%. CONCLUSION: We conclude that biofilm bacteria are much more resistant to CPFX than the floating bacteria. The combination of CPFX and CAM, ROX HA were more effective in decreasing the biofilm bacteria survial rate than CPFX alone. The results suggest that the combination of CPFX and CAM, ROX, HA can be one of the therapeutic manauvers against biofilm bacteria. PMID- 12548949 TI - [Isolation, purification and bioactivities of exopoly saccharides from fermented broth of Ganoderma lucidum]. AB - The exopolysaccharides of Ganoderma lucidum(GLEP) extracted from the fermentation broth after removing protein by Sevage and protease digestion procedures, were applied to a column of DEAE-cellulose(OH- form), and eluted stepwise with distilled water, sodium hydrogen carbonate (0.1 mol/L, 0.3 mol/L, 0.5 mol/L successively) and 0.1 mol/L sodium hydroxide. Five fractions were obtained, and the main fraction was known as GLEP-I, furthermore subjected to chromatography on a column of SepharoseC1-6B, eluted at a flow rate of 30 mL/(cm2.h), the relative viscosity of sample solution of 1.5. Two fractions, GLEP-IFr1 and GLEP-IFr2 with a ratio of 3.8:1, were obtained. Molecular weight of GLEP-IFr1 and GLEP-IFr2 was estimated to be 38,000 and 22,000 Dalton respectively by Membrane Osmometer. The animal test showed that GLEP-IFr1 could inhibited the growth of Sarcoma 180 tumor in mice. The average inhibition ratio was 57.4% (i.p. 10 mg/kg for 10 days). The result of immunological activity showed that GLEP-IFr1 could significantly improve macrophage cytophagy. PMID- 12548950 TI - [Detection of Neisseria gonorrhoeae(NG), Chlamydia trachomatis(CT) and Ureaplasma urealyticum(UU) by multiple primer PCR]. AB - For the detection of Neisseria gonorrhoeae(NG), Chlamydia trachomat is(CT) and Ureaplasma urealyticum(UU) in urogenital infection in one specimen, multiple primer PCR was used in 156 sex transmitted disease patients, it was shown that in 156 specimens there were 40 NG positive and 31 CT positive and 27 UU positive, 10 NG + CT positive, 4 NG + UU positive, 3 CT + UU positive and 2 NG + CT + UU positive respectively. The results were in correspondence with each other. It is suggested that multiple primer PCR is a quick, sensitive, simple and specific technique which can be applied in clinical test. PMID- 12548951 TI - [History and prospects of the research on mutanolysin]. PMID- 12548952 TI - [Genetic diversity of rhizobia of Medicago edgeworthii by AFLP and RFLP analysis]. AB - Two hundred and ninety one isolates of rhizobia sampled from root nodules of Medicago edgeworthii which came from Yun Nan Province in china were analyzed by amplified restriction fragment polymorphism(AFLP) technique. According to the AFLP banding patterns, the results showed most of these isolates were genetically diversity. Ninety isolates were selected as representing the diversity among the 291 isolates. The 90 isolates were clustered into three groups at the level of similarity of 79% by computer analysis of the data. Additional representative isolates and reference strains were identified by restriction fragment length polymorphism (RFLP) analysis of PCR-Amplified 16S rDNA. Two different 16S rDNA PCR-RFLP types were found, which indicated that the isoltes were phylogentically closely related to Rhizobium mongolense. PMID- 12548953 TI - [Detection of cucumber mosaic virus subgroup II isolate from tobacco plants in Yunnan province]. AB - One of the spherical virus(designated as CMV-YNb) was isolated form the tobacco plants in Yunnan province. SDS-PAGE showed that this virus contained one polypeptide with molecular weights of 55 kD. The polypeptide was submitted to automated Edman degradation, and found the 10 amino acid sequence of the N terminus of 55 kD protein shares 100% identity with amino acid sequence of coat protein of CMV-Q, which is belong to CMV Subgroup II. Western blot analysis also showed that this 55 kD protein was serological related to CMV-Q. A 0.9 kb cDNA fragment was amplified by RT-PCR from the RNA template of CMV-YNb with the QCf/QCr primer pair, which was designed according to the CP gene sequence of CMV Q. The cDNA fragment was then cloned and sequenced. The fragment comprises 865 bp (EMBL accession number: AJ242585), including a 197 bp of 5' intergenic region, 657 bp of CP gene encoding 158 amino acids and a 11 bp of 3' partial non-coding region. Comparison of the nucleotide and amino acid sequences of CMV-YNb CP gene with that of CMV strains belonging to Subgroup II, and that of CMV strains belonging to subgroup I showed 97%-98% and 96%-99%, 75%-76% and 78%-79% identities, respectively. With these data, we made a conclusion that the CMV-YNb is an isolate of CMV Subgroup II. PMID- 12548954 TI - [Development of a system for Polymyxa graminis infection and transmission of wheat yellow mosaic virus]. AB - In growth chambers, a system was developed for infection of Polymyxa graminis to wheat(Triticum aestivum L.), which is an obligate parasite. By the system, the fungus could be propagated in large quantity rapidly and the life cycle of P. graminis in wheat was observed in a period of 13 to 15 days. The methods for separating P. graminis as single cystosori from cultured wheat roots or dried roots were simplified. By tests for the factors affecting on P. graminis infection, including inocula types, seedling age of host plants, temperatures, pH value and nutrient contents etc., the system was modified to more perfection. With an integration of optimal temperature and other elements in the system, which affect on infection and development of P. graminis, wheat yellow mosaic virus (WYMV) was transmitted constantly and efficiently by the fungal vector. In average, 70% of wheat plants inoculated by the viruliferous fungi could infected by WYMV and the typical symptom of wheat mosaic disease would appear at 30 days post-inoculation mechanically. PMID- 12548955 TI - [Overproduction and partial characterization of the Ssh7a and Ssh7b proteins from Sulfolobus shibatae]. AB - The Sulfolobus shibatae Ssh7a and Ssh7b proteins have been separately overproduced in Escherichia coli and purified using a simple procedure which includes a heat treatment step. The recombinant and native Ssh7 proteins are similar in the ability to bind both negatively supercoiled and relaxed DNAs. In addition, the recombinant Ssh7a resembles the native Ssh7 protein in constraining negative DNA supercoils. Our data suggest that the two isoforms of Ssh7 interact with DNA in a similar fashion, and the methylation state of Ssh7 interact with DNA in a similar fashion, and the methylation state of Ssh7 does not affect DNA binding and supercoil constraining by the protein. PMID- 12548956 TI - [Characterization of the insecticidal crystal protein genes of Bacillus thuringiensis YBT-1520]. AB - The insecticidal crystal protein genes of highly toxic Bacillus thuringiensis YBT 1520 were analyzed. Southern hybridization and PCR analysis indicated that this strain bears cry1Aa, cry1Ab, cry1Ac and cry2 genes. The copy number of three cry1A genes is different and cry1Ac gene (here named cry218) is the highest. The restriction location of three genes is different from that in typical strains, such as HD-1 and HD-73 strains. 4190 bp nucleotide sequence of cry218 gene was determined and this gene was named cry1Ac10 in the recently published gene nomenclature. PMID- 12548957 TI - [Identification of cry-type genes of 31 Bacillus thuringiensis isolates and analysis of their expression product]. AB - In this study, cry-type gene of 31 isolates of Bt had been identified using PCR RFLP identification system, and SDS-PAGE analysis and bioassay had been performed. 25 strains containing cry1 type gene express 130-150 kD protein and 16 of them contain cry1I gene, which codes 80 kD protein and is larvicidal to both lepidopteran and coleopteran species. 15 strains contain both cry1 and cry2 type gene; 10 strains contain unknown cry-type gene; 6 strains don't contain seven identified cry-type gene. Result of bioassay shows that the strains containing both cry1 and cry2 gene have high toxicity to lepidopteran order, 7 of them are strongly insecticidal to Pristiphora beijingensis and Lymantria dispar larvae; the strains containing cry1Ab + 1Ac + 2A or cry1Aa + 1Ac + 2A have high activity to Heliothis armigera; no cry gene, no activity. It is indicated that identification of cry-type gene and its analysis of SDS-PAGE protein can predict insecticidal activity of Bt isolates, and it is no pertinency between cry-type gene and serotype of Bt strain. PMID- 12548959 TI - [Cloning and expression of metallothionein gene of Bombyx mori]. AB - Yeast MTI gene from vector pCMI-1 was used as a probe. It appeared strong hybridization signals when the total DNA of Bombyx mori Huishu eggs hybridizes with the probe. The 1-6 kb DNA fragments were isolated from the EcoR I digested total DNA of Bombyx mori Huishu eggs and ligated with M13- vector digested by restriction EcoR I. The ligation mixtures were used to transform E. coli DH5 alpha. blue/White colonies selection was used to identify colonies with insert. Approximately 4000 white colonies were selected, so the part genomic library of Bombyx mori was constructed. Three positive colonies were gained from the genomic library by southern blotting analysis, designated T1 (pZHC-1), T5 (pZHC-5), T7 (pZHC-7). Digesting the recombinant plasmid pZHC-5 with 12 restriction enzymes, the results suggested that the inserted fragment was about 1.2 kb and there was only a Hind III site. The experiment of resistant to CuSO4 proved that the DH5 alpha cells contain recombinant plasmids were more resistance than the recepient DH5 alpha cells. According to these results, the inserted fragment possibly contains the gene encoding Metallothionein of Bombyx mori. The sequence analysis of the inserted fragment and its high-expressions in E. coli are in progress. PMID- 12548958 TI - [Molecular cloning of enhancin gene from Helicoverpa armigera granulosis virus and its expression in E. coli]. AB - In order to provide recombinant enhancin for studying its mechanism of increasing the mortality of larvae infected by HaNPV and creating new insecticide, enhancin gene from Helicoverpa armigera granulosis virus was amplified by PCR technique. 2.7 kb fragment of enhancin gene was cloned into EcoRI/XbaI site of plasmid pBluescript KS. Sequence analysis revealed that enhancin gene was similar with that reported in the literature except eight nucleotides and five amino acids. Thereby enhancin gene was inserted into vector pET-30a and expressed successfully in Escherichia coli BL21(DE3). The preliminary bioassay of expressed product and HaNPV indicated that mortality of larvae increased 31.7%-34.1% in 7 post infection days and the LT50 decreased at least 1.5-2.1 days. PMID- 12548960 TI - [Preparation of monoclonal antibodies against malaria through immunization of mice with recombinant plasmid DNA]. AB - BALB/c mice were immunized by injection of recombinant plasmid DNA carrying MSP1 31 fragment of Plasmodium falciparum. After antiserum was induced, McAbs were prepared by fusing SP2/0 myeloma cells with spleen cells. Specific McAbs were detected in hybridoma culture supernatant and mouse ascites by ELISA, positive hybridomas were cloned by technique of limiting dilutions. Two McAbs, 9H9 and 8A2, against the malaria protein were produced and characterized. The titers of the two McAbs in acites were 1:10,000 and 1:2500 respectively. The Ig subclass of these McAbs was IgG1. Western blot test showed that the McAb specifically reacted with MSP1-31 antigen. These results proved that monoclonal antibodies could be produced by use of immunization of mice with plasmid DNA or DNA vaccine. PMID- 12548961 TI - [Screening DPPH radical scavengers from Monascus sp]. AB - A strain belongs to Monascus sp. was separated from hongqu(Chinese beni-coji) which was used to brew red wine in Changxin county, Zhejiang province, and its DPPH radical scavenging metabolites were studied. Results showed that the yield and radical scavenging activity of neutral extract with EtOAc were higher than that of acid and basic extract. After shaken at 30 degrees C, 100 r/min for 5 days, this strain produced largest amount of neutral extract. Further partitioning the neutral fraction with silica gel column chromatograph, LH-20 column chromatograph, MPLC and HPLC, we screened 15 free radical scavengers whose yield was more than 2 mg and purity was higher than 85% in HPLC (260 nm absorption). Several of them were analyzed with 1H-NMR and 13C-NMR spectrum from which we concluded that B1-3 and C3-1-7 were two kinds of substitute of benzene, and C3-1-7 may be 3-OH-4-OCH3-benzoic acid. The scavenging activity of 40 mumol/L B1-3 was about 65%, and that of C3-1-7 was lower than 56%, while as the control the scavenging activity of 40 mumol/L Vc and Ve was 80.4% and 78.4% respectively. PMID- 12548962 TI - [Isolation and characterization of mutants of Aspergillus niger deficient in extracellular proteases]. AB - Several mutants of Aspergillus niger 3.795, deficient in extracellular protease expression, have been isolated and characterized biochemically after in vitro UV mutagenesis of conidiospores and selected by halo-screening on the gelatin/casein plate and skim milk plate. Extracellular proteolytic activities of 3.795-1-23 and 3.795-1-30 are 5.4% and 8.4% of the parental strain and these strains may be used as the host for efficient expression of foreign proteins. PMID- 12548963 TI - [A procedure for developing selective isolation of Pseudonocardiacetes from soil samples]. AB - Pseudonocardiacetes are a group of rare-actinomyces with type IV cell walls and without mycolic acids. Five different selective isolation media S1, S2, S3, S4 and S5 were devised and practiced to improve their selective isolation efficiency. The media were detected by 33 type strains of Pseudonocardiaceae, 4 strains of Saccharothrix and 15 type strains of Streptomyces cultured on them. Soil samples were examined on media S1 ad S2 compared with glucose-asparagine agar. It shows that media S1 and S2 are effective for growth and isolation of Amycolata, Pseudonocardia, Saccharomonospora and other genera of Pseudonocardiaceae. PMID- 12548964 TI - [L-lactic acid fermentation by immobilized Rhizopus oryzae in a three-phase fluidized-bed]. AB - L-lactic acid fermentation in a three-phase fluidized-bed bioreactor was studied by using immobilized Rhizopus oryzae cells with polyurethane particles as supporter. The results indicated that the immobilized cells were stable after used for 8 batches repeatedly. The rate and the yield of lactic acid fermentation with glucose as carbon source were 19.1 g h-1.L (bead)-1 and 82.5%, respectively. With the hydrolyzate of corn cob residue from xylose manufacturer by cellulase hydrolysis, the yield of lactic acid reached 71.9%. PMID- 12548965 TI - [Purification and characterization of an endo-beta-1, 4-mannanase from Bacillus subtilis BM9602]. AB - An extracellular neutral endo-beta-mannanase(endo-beta-1, 4-D-mannan mannanohydrolase, EC 3.2.1.78)of Bacillus subtilis BM9602 was purified to electrophoretic homogeneity by ammonium sulfate precipitation and DEAE-cellulose DE22 chromatography with 45.5 fold and 5.9% yield. It's molecular weight and pl value were 35 kD by SDS-PAGE and 4.5 by isoelectric focusing, respectively. The enzyme was the most active at pH 5.8. The optimum temperature of the enzyme activity was 50 degrees C. The enzyme was stable at pH 6.0-8.0 and below 50 degrees C. The activity of the enzyme was inhibited by Hg2+, Ag+ strongly. For various substrates, such as locust bean gum, guar gum, sesbania gum and konjac gum, Km and Vmax value of the enzyme were 3.8, 14.9, 11.3, 2.4 mg/mL, and 24.5, 86.5, 38.4, 19.8 mumol.min-1.mg-1, respectively. The enzyme hydrolize various plant beta-mannans, with valuable oligosaccharides and without monosaccharide. PMID- 12548966 TI - [Studies on biosorption of Au3+ by Bacillus megaterium]. AB - Some characteristics of Au3+ biosorption and bioreduction by Bacillus megaterium D01 biomass were reported. The optimum pH value of Au3+ biosorption by strain D01 was 3.0. The biosorption was a rapid process, reaching 95% of the maximal capacity within the first 5 min of contact. Temperature did not affect the biosorption. The biosorptive efficiency(99.1%) and capacity (302.0 mg Au3+/g dry weight biomass) were achieved under the conditions of pH 3.0 and 30 degrees C, both concentration with an initial Au3+/biomass radio of 305 mg/g dry weight for 30 min of the contact. The D01 biomass could reduce Au3+ in the solution to Au0, Au0 on the cells surface and in the contact solution could be crystallized to different forms gold crystals, Au3+ impregnated on SiO2 and alpha-Fe2O3 could be reduced to Au0 by D01 biomass. The results of electrochemical reaction indicated that the strain D01 exhibited a high selectivity for Au3+. PMID- 12548967 TI - [Purification and properties of degrading enzyme of organophosphorus pesticides from Aspergillus ornatus Z58]. AB - Degrading enzyme of organophosophorus pesticides was purified from Aspergillus oryzeZ58 by (NH4)2SO4 precipitation, Sephadex G-100 gel filtration, DEAE cellulose chromatography with 34.2-fold purification, 17.8% recovery and 76.2 U/mg specific activity. The purified enzyme was homogeneous on polyacrylamide gel electrophoresis. The molecular weight of the enzyme estimated with Sephadex G-100 was 67000. The optimal conditions for activity were pH 7.2, temperature 45 degrees C. The enzyme was stable over the range of pH 6-10 and below 40 degrees C, the activity was inhibited by SDS and stimulated by Cu2+, the substrate ranges were nonspecific to organophosophorus pesticides. PMID- 12548968 TI - [Transfer of two large linear plasmids pHZ1000 and pHZ1001 by conjugation from Streptomyces T8-4 to S. lividans ZX1]. AB - Using pulsed-field gel electrophoresis (PFGE), two indigenous large plasmids were isolated from Streptomyces T8-4. Two dimentional PFGE revealed that both plasmids were linear molecules. By parallel electrophoresis with linear plasmids of known sizes, the two linear plasmids were estimated to be approximately 230 kb and 90 kb, which were designated as pHZ1000 and pHZ1001, respectively. Both plasmids could be transferred into S. lividans ZX1 by conjugation, which is detectable by "pock" formation. Five S. lividans ZX1 derivatives which carry one or two plasmids were isolated and characterized by Southern hybridization and PFGE. PMID- 12548969 TI - [Screening and fermentation conditions of a bacterial strain over-producing xylanase]. AB - A xylanase producer WXULI-11 was isolated and identified as Pseudomonas sp. By combination treatment of WXULI-11 with UV and NTG, a mutant WLUN024 was obtained and its enzyme activity reached 354 IU/mL in 36 h by shaking flask. The mechanism of its xylanase production was primarily studied. The results showed that this enzyme was induced by xylan, xylose and some other xylosic material, while xylose had "two-way effect" on xylanase synthesis. Effects of nitrogen and other factors on enzyme production were also investigated. PMID- 12548970 TI - [Study on whi-genes related to differentiation of Streptomyces spp]. PMID- 12548971 TI - [Pathogenic mechanism of entomopathogenic bacteria--Xenorabdus and Photorhabdus]. PMID- 12548972 TI - Alleviation of pre-exposure to low-dose 16O8+ ion on mouse testicular histological damage induced by subsequent high-dose irradiation. AB - The testes of the B6C3F1 hybrid strain mice were irradiated with 0.05 Gy of 16O8+ ion as the pre-exposure dose (D1), and were then irradiated with 2 Gy of 16O8+ ion as challenging radiation dose (D2) at 4 h after per-exposure. Testicular morphology was observed by light microscope at 35th day after radiation. The results showed that irradiation of mouse testes with 2 Gy of 16O8+ ion significantly impaired, mainly reduction of tubule diameter and decrease or loss of germ cells in various developing stages, especially spermatogenic elements. Pre-exposure to a low-dose (0.05 Gy) of 16O8+ ion significantly alleviated above mentioned damage on testicular morphology induced by subsequent a high-dose (2 Gy) radiation. PMID- 12548973 TI - [Expression of VEGF on rabbit skin fibroblasts using retroviral vector]. AB - A replication-deficient recombinant retrovirus containing the cDNA coding for human vascular endothelial growth factor (VEGF) was generated, and then infected rabbit primary skin fibroblasts. After selection with G418, the transduced colonies have the ability of producing VEGF. The integration and expression of VEGF in transduced cells were confirmed by Southern blot, PCR, Northern blot and RT-PCR assay. The VEGF secreted by transduced cells has strong bioactivity when assayed by endothelial proliferation and Miles vascular permeability assay. Thus, this study pave the way for future study of biological and physiological effect of VEGF in vivo. PMID- 12548974 TI - [Influence of magnetic field on nitric oxide in hypothalamus and its relation to hypothalamic neuroendocrine nuclei]. AB - By employing nitric acid reductase-spectrophotometry and NADPH-diaphorase/AVP cytochemistry technique, the effects of magnetic field on NO in hypothalamus and relations to Paraventricular Nucleus (PVN), Periventricular Nucleus (PEN), Supraoptic Nucleus (SON) and Suprachiasmatic Nucleus (SCN) were investigated. It was found that the NADPH-d positive neurons and some NADPH-d/AVP dually stained neurons existed in PVN, PEN, SON, but not in SCN, and the magnetic field induced NO (OD) increase there and the high NO (OD) level lasted for 3 hours. The results suggested that NO (OD) increase after the treatment of magnetic field in hypothalamus may result from strong expression of NOergic neurons in the PVN, PEN and SON. The coexistance of NO and AVP may play important role in the regulation of endocrine and neuroendocrine by the magnetic field. PMID- 12548975 TI - [Cloning and tissue expressive pattern analysis of the human ribosomal S6 kinase RPS6KA5 cDNA]. AB - Human ribosomal protein S6 kinase includes two protein families: P90RSK and P70S6K, they participate in two different signaling pathways. When the two kinases were inhibited by their antibodies or rapamycin, the proliferation of cells was arrested. However, their analog, the immunosupressant FK-506, can inhibit the proliferation of fibroblast PBL1 without interfering with the activities of P90RSK, P70S6K and MAPK. We take the tactics of "homolog screening" to demonstrate whether there are some novel proteins which can substitute for the known P90RSK and P70S6K or other pathways without interfering with the known P90RSK and P70S6K. With the conserved sequence of mouse p90RSK as a probe, we screened the homologous sequence in NCBI EST database and got three human EST fragments. With the assembled contig as a probe to screen human brain cDNA library, a full-length cDNA of 3833 bp was attained. It contains a completed open reading frame from 165 bp to 2570 bp encoding 802 amino acids. The putative protein has higher homology with other members of p90RSK family. The gene was named RPS6KA5, the accession number in GenBank is AF090421. Northern hybridization showed the gene expressed in 16 human tissues tested, and the gene was localized in 14q31-32.1 by RH mapping. Another novel P70S6K gene has also been cloned. Thus, our initial presumption that there is an analog of known P90RSK and P70S6K in human beings was proved. PMID- 12548976 TI - [Extension of life-span of normal human fibroblasts by reconstitution of telomerase activity]. AB - Most of normal human somatic cells can divide only a finite number of times and inevitably become senescent. Telomerase is an enzyme that imparts replicative immortality by maintaining the length of the telomeres when expressed in reproductive and cancer cells. Cells that are mortal do not express the telomerase. Recently it was reported that the life-span of the normal human cells could be successfully extended by introduction of telomerase into these cells. We have found, in the previous work, that fibroblasts exhibited an osteogentic potential, and therefore, can be considered as a type of "seed cells" in tissue engineering for bone repairing and reconstruction. But this potential was impaired by the limitation in life-span and proliferative capacity of the normal fibroblasts. In the present work, plasmid pGRN145 bearing a cDNA insert of human telomerase reverse transcriptase (hTERT) was introduced into the fibroblasts with osteogenic potential by electroporation. The stable hTERT+ fibroblast clones was established and cultured for long-term in a medium containing hygromycin-B. The exogenous hTERT mRNA expression and telomerase activity were detected. The hTERT+ fibroblasts showed shorter population doubling time and no beta-galactosidase stain, which indicated a stronger proliferative capacity and fewer signs of cell senescence, compared to their hTERT- counterpart. These evidenced that the life span of hTERT+ fibroblasts was extended. The assays for DNA euploidy by flow cytometry and chromosome karyotype by cytogenetic technique showed no signs of heteroploidy, providing the data for cell carcinogenesis and utilization safety. The results of the present study suggested that the introduction of hTERT could make the life-span of normal fibroblast extended without causing their malignant transformation, and such type of "longevous" fibroblasts might be clinically useful in tissue engineering for bone repairing and reconstruction. PMID- 12548977 TI - [The different effects of CaM inhibitors of phenothiazines on the proliferation of Saccharomyces cerevisiae and Schizosaccharomyces pombe]. AB - Low concentration of phenothiazines apparently stimulated the proliferation of S. pombe, the cell density incubated for 54 hours by preincubating the cells with 20 mumol/L trifluoperazine (TFP) in the EMM-Ca medium was two times more than the control. The stimulation was more obvious with lowing the concentration of calcium in the culture medium, TFP cooperated and complemented with calcium in stimulating the proliferation of S. pombe. When the original inoculated cell density was 5 x 10(6) cells/ml or during the logarithm period of growth curve, the proliferation of S. pombe wasn't affected by the low concentration of TFP. While when the concentration of TFP was increased to 100 mumol/L, the promotion effect of TFP on proliferation of S. pombe declined obviously and the proliferation of S. pombe was inhibited completely when TFP up to 200 mumol/L. The cell proliferation also could be inhibited by CaM antagonist W7 and W7 agarose, the inhibition was increased with increasing the concentration of antagonist. On the other hand, 20 mumol/L TFP used by the same method as above arrested the cell division cycle of Saccharomyces cerevisiae at a single G2 + M nuclei stage, the cells was penetrated easily by TFP, the fluorescence in cells was very obvious when TFP was 20 mumol/L, but it was difficult to penetrat by TFP in the cells of S. pombe and the Ca2+ influx of S. pombe could be induced rapidly by 20 mumol/L TFP. In this article, the cause of different effects of TFP on cell proliferation of S. pombe and S. cerevisiae was discussed, it was due to the difference of penetration of TFP and stimulation by calcium in the two kinds of cells. PMID- 12548978 TI - [Isolation and sequence analysis of a mitochondrial DNA fragment associated with CMS in Hong Lian type rice]. AB - A CMS line specific mtDNA band TA1 was isolated by comparing the mtDNAs of CMS line and maintainer line using AFLP technique. The Northern blot analysis of the TA1 mtDNA fragment show that it transcripts only one RNA molecular each in CMS line, maintain line and the first filial generation, but the RNA moleculars are different among each other. By sequencing, the fragment is found to be 202 base pairs in length. It contains code ATG, ATT, AGA and AGG, two small repeated sequences 5'TGTAC3' and 5'ATTATTTT3' and one small inverted-repeated sequence 5'GGGAAACA3'. The results imply that the mtDNA fragment may be a coding region and may be relative to CMS of Hong Lian type rice. PMID- 12548979 TI - [Expression of thermostable alpha-amylase gene in potato]. AB - The thermostable alpha-amylase gene cloned from Bacillus lichemiformis was reconstructed into an expression vector pAMY721M under the CaMV35S promoter. The vector was transferred into A. tumerfaciens ABI. The thermostable alpha-amylase gene was transferred into Solanum tuberosum L. via Agrobacterium mediation according to the revised method of ZHAO Shu Juan et al (1997) and YANG Mei Zhu et al (1992). Shoots were induced and regenerated on MS medium with 2 mg/L ZT, 0.1 mg/L IAA and 100 mg/L kanamycin. Putative transformants were selected with kanamycin and roots induced on MS medium with 0.15 mg/L IAA. PCR analysis and thermostable alpha-amylase activity assay were done to identify the transgenic plantlets. Among them, 102,001, 102,607, and 110,402 were showed to have a higher alpha-amylase activity than untransformed control. PMID- 12548980 TI - [Plant regeneration from protoplast culture in flax (Linum usitatissimum)]. AB - Shoot protoplasts of four fiber flax (Linum usitatissimum) varieties (7309, 948, Belinka and Viking) were isolated and cultured. The optimal condition for higher protoplast yield 1.8 x 10(6)/gFW and activity 85.5% (c.v. 948) were from 10 day old seedings. Culture in V-KM Agroase-island medium led to first divisions after 3 days (c. v. 948), and after twenty days with an efficiency of 36% of divided cells and 5.2% in plating efficiency. Plant regeneration was obtained in 7309 and Belinka on agar media B5-2, MS3 containing 0.6 mg/L 6-BA and 0.1 mg/L NAA. Roots and leaves regeneration were observed in Viking and 948 respectively. PMID- 12548981 TI - [Immunocytological localization of IAA in the parenchyma cell and vascular elements in the graft union of Cucurbita pepo/Cucurbita moschata at the early developmental stage]. AB - Immuno-gold localization of IAA in cells of the graft union in the explant internode graft of Cucurbita pepo/Cucurbita moschata were investigated with electron microscopy. In parenchyma cells near the graft union, the gold particles were mainly accumulated in nucleus, plastid and endoplasmic reticulum, while no gold particles was detected in Golgi body, mitochondrion, cell wall and vacuoles. In the differentiating xylem element, the gold particles were labeled in secondary wall and cytoplasm. In the sieve element gold particles were found in the sieve plate, sieve pore and cytoplasm. There was a dense label of the gold particles in the companion cell. The role of IAA in the differentiation of the vascular elements was discussed. PMID- 12548982 TI - [Effects of IAA and stimulated microgravity on formation of adventitious roots of Chinese cabbage]. AB - Under the induction of indole-3-acetic acid (IAA), adventitious roots were differentiated on hypocotyl segments derived from seedlings of Chinese cabbage (Brassica campestris spp. pekinensis). IAA at concentration of 0.4-1.0 mg/L in solid MS medium incited many adventitious roots on hypocotyl segments. The earliest anatomic changes were observed on cut surface of hypocotyl segments under optical microscope 24 hours after IAA treatment: cytoplasmic and nuclear density became higher in a few of parenchytmatous cells adjacent to phloem in tissue of pericycle, followed by cell divisions. Lately, the dividing cells expanded and developed into root primordium from which root cap was differentiated. After five days, most roots protruded through hypocotyl cortex and appeared just below the cut surface. The rooting capacity of the segments derived from three regions of each hypocotyl was different. High level of IAA modified the polarity of root formation on segment inserted upside down and sucrose increased the function of IAA. Additionally, microgravity did not significantly change the rooting polarity under the condition of stimulated microgravity, but it increased the competence of explants to IAA treatment. The results presented here provided an experimental system for further investigation of molecular events associated with adventitious root initiation. PMID- 12548983 TI - [A new method for extracting the dinoflagellate crypthecodinium cohnii chromosomes and observation of their ultrastrcture]. AB - In this paper, a new method of extracting the dinoflagellate Crypthecodinium cohnii chromosomes was proposed. The ultrastructure of dinoflagellate Crypthecodinium cohnii chromosome was studied by transmission electron microscope. The chromosome fibres of dinoflagellate fold in a different manner from that of typical eukaryotes. PMID- 12548984 TI - [Studies on the photosynthetic characteristics of wheat variants by introduction sorghum DNA]. AB - Spring wheat Longchun No. 13 and Ganmai No. 8 were introduced by sorghum DNA via pollen tube pathway, and two stable progenies (89,122 and 89,144 respectively) were obtained through selection for several years. The results of photosynthetic analysis showed that photosynthetic rate (P) and photosynthetic/transpiration rate (E/P) of 89,122 and 89,144 were mediated between their recipients and donor respectively, which demonstrated stronger photosynthetic efficiency than their receptors. However, the delta 13 C values in assimilates from 89,122, 89,144 and their maternal wheat cultivars G8 and L13 were approximately same, and they were significant different from sorghum. The chlorophyll content and the ratio of Chla/Chlb of them also showed that wheat cultivars and sorghum were different. It might demonstrated that the stronger photosynthetic efficiency of the progenies was due to their higher Calvin cycle metabolism efficiency rather than their carbon-fixation pathway difference. On the other hand, it also suggested that the distant genetic materials of the sorghum affected the photosynthetic characteristics of 89122 and 89144. PMID- 12548985 TI - [Phenotypic features and genetic diversity of strains of Rhodopseudomonas]. AB - 14 strains of purple non-sulfur bacteria and 3 type strains of Rhodopseudomonas were investigated by phenotypic characterizing, DNA-DNA hybridizing and DGGE analyzing. The characteristics of their morphology, structure and physiology show that 14 strains of isolates have the features of the genus Rhodopseudomonas: dividing asymmetrically by budding, lamellar intracytoplasmic membranes, presence of bacteriochlorophyll a and carotenoids of the normal spirilloxanthin series. On the base of their differences in morphology, growth, carbon sources 14 strains can be divided into 2 groups: g-c group, T group. Parts of 16S rRNA gene were amplified by using a pair of primers: 341f-906r. Denaturing gradient gel electrophoresis(DGGE) of PCR-amplified fragment of rDNA indicated that they have 3 genetic types: 1 of them, including 6 of 8 strains of T groups is similar to that of Rps. rutila R1, the other 2 types, represented separately by 2 of T group or all the g-c group, were different from that of either of 3 type strains The Data of the DNA-DNA hybridization of 8 representative strains indicated that the differences between different types were enough to separate them into different species including 2 possible new species. PMID- 12548986 TI - [Study on the morphological processing of classical swine fever virus in cultured cells]. AB - An infected mode between the Thiverval strain and Chinese strain of CSFV and MPK cells are established. The morphological structure and processing of different strains(T strain, C strain and F strain) of CSFV were studied by ultra-thin section electron microscopy. The virions of CSFV are roughly round and approximately 70 nm in diameter with a 40 nm core, and are wraped by membrane. The distinctive pathway of the maturation and release of CSFV are observed. Virons on different deverloping state and the pathologic changes of host cells have also been reported in this paper. PMID- 12548987 TI - [Molecular cloning and expression of E2 gene of the Chinese classical swine fever virus(shimen strain) and preliminary studies of its DNA vaccine]. AB - A 1.1 bp fragment of E2 gene of Chinese classical swine fever virus(CSFV) Shimen strain, a standard virulent strain, was amplified by RT-PCR from total RNA of cell cultures infected by CSFV, and cloned into pGEM T vector. The nucleotide sequence of this fragment was sequenced by Sanger's method and the amino acid sequence was deduced. Compared with the corresponding region of Alfort, Brescia and C strain of CSFV, the nucleotide sequence homology is 84.7%, 92.6% and 95.2% respectively, and the amino acid sequence 89.4%, 92.6% and 94.6%, respectively, we subcloned 1.1 bp of E2 gene cDNA into baculovirus transfer vector and successfully constructed two recombinant baculoviruses expressing GST-E2 and GST GFP-E2 fusion protein respectively by homologous recombination in sf-9 cell. Furthermore, we also constructed recombinant eukaryotic expression vector pcE2 containing E2 gene in frame and transfected COS-7 cell by lipofectamine, the indirect immunofluorescence assay (IFA) showed that the expressed E2 protein can be recognized by E2 specific monoclonal antibody the pcE2 DNA was directly injected into BALB/c mice intramuscular(i.m.) and the CSFV E2-specific antibodies was measured by enzyme-linked immunosorbent assay(ELISA) the ELISA results indicated the E2-specific antibodies was induced in inoculated mice and virus neutralization assays also indicate single inoculations of plasmids expressing CSFV E2 glycoprotein raised neutralizing antibody in BALB/c mice. these results will be beneficial to investigate the possibility of DNA vaccine against CSFV. PMID- 12548988 TI - [Infection of white spot baculo-like virus(WSBV) in Penaeus chinensis: evidence from electron microscopy and DNA hybridization]. AB - Severe epizootic causing high mortalities in cultivated Penaeus chinensis has occurred every year in mainland China since 1993 with great economic loss. Naturally diseased P. chinensis sampled in Qingdao shrimp farming regions between 1993 and 1998 were examined by electron microscopy. A similar non-occluded nuclear bacilliform virus like the white spot baculo-like virus (WSBV) reported in Taiwan was mostly found in the gill, cuticular epidermis under the exoskeleton, stomach, lymphoid organ and other tissues of ectodermal and mesodermal origin from these diseased samples. The viron was enveloped with a mean size of (125 +/- 7.6) nm x (345 +/- 16) nm. Sometimes, a nipple-like appendage could be found protruding from one extremity of the viron. Envelopes were all lost after purification from cesium chloride gradient ultracentrifugation. The cylindrical nucleocapsid in negatively staining preparations was (80 +/- 13) nm x (380 +/- 24) nm with 13-16 stripes perpendicular to the longitudinal axis of the nucleocapsid. Based on its pathogenic and morphological characteristics, this virus should be related to WSBV. A pair of primers created from information of WSBV genome DNA SalI fragment produced a 355 bp band using the WSBV isolate from P. chinensis in mainland China, as the DNA template. The specific PCR product was cloned, sequenced and labeled with digoxigenin (DIG) DNA labeling kit (Boehringer Mannherm). The sequence of the probe is identical to that of WSBV. All of the sampled diseased shrimps between 1993 and 1998 reacted with the DIG-labeled probe by dot-blot hybridization and no hybridization was observed using DNA from health shrimp as templates. These results based on electron microscopy and DNA hybridization indicated that the shrimp epizootic occurred in mainland China was related to the WSBV infection previously occurred in Taiwan early 1992. PMID- 12548989 TI - [Identification and functional analysis of NIFA from Mesorhizobium huakuii 159]. AB - The nifA gene from Mesorhizobium huakuii 159 was identified and sequenced. An open reading frame coding for 44734D. NifA protein would detected. Comparison of the deduced amino acid sequence with other NifA proteins revealed the presence of the conserved central domain and the C-terminal DNA-binding domains. But the predicted M. huakuii NifA protein lacked a N-terminal domain that is present in the homologous proteins. Mutants with Nif- phenotype were obtained by site directed Tn5 mutagenesis. A nifA constitutively expressed plasmid was constructed, and this plasmid could activate the nifH expression in YMC11. PMID- 12548990 TI - [A new resolution vector with cry1Ac10 gene based on Bacillus thuringiensis transposon Tn4430]. AB - A new resolution vector with cry1Ac10 gene based on TnpI-mediated site-specific recombination system of Bacillus thuringiensis(Bt) transposon Tn4430 was developed. The gene cry1Ac10, encoding a protoxin against plutella xylostella larvae, and the gene ori1030, from a plasmid of wide type Bacillus thuringiensis, were inserted into two copy sets of RES sites, named pBMB801. When pBMB801 was introduced into crystal negative Bt host BMB171, antibiotic resistance genes and other non-Bt DNA can be selectively eliminated. This recombinant plasmid was found very stable without antibiotic selection. The resulting strain only contained Bt DNA and is free of antibiotic resistance genes. This strategy should facilitate regulatory approval for its development as a commercial biopesticide. PMID- 12548991 TI - [Selection and recombinant expression of an immunodominant B cell epitope of HIV gp41]. AB - A novel method was designed for disease-specific B cell epitope mapping and epitope expression in E. coli. A phage library displaying random dodecamers was biopanned first with human total IgG antibodies against HIV-1, and then non specific phages were subtracted by HIV(-) polyclonal antibodies. After three rounds of screening, the positive phages were tested in an ELISA for their reactivity with HIV(+)-IgG and HIV(-)-IgG antibodies. Phages that showed positive reactivity with HIV(+)-IgG, but negative to HIV(-)-IgG, were selected and their displayed peptides were determined by DNA sequencing. All the 13 positive clones sequenced displayed five kinds of peptides (SPKCLGKLLCAF, THQCLGKLQCGV, SCSAKFTCTTQI, KSDCSARFMCSV, DCLKQWACEWSR) that have homology to the HIV-1 gp41(602GCSGKLICTTNV613), demonstrating there is a dominant epitope in the region. PMID- 12548992 TI - [Gene cloning and expression of prolyl endopeptidase from Aeromonas punctata]. AB - Prolyl endopeptidase gene was cloned from Aeromonas punctata subsp. Punctata(ST 78-3-3) using activity screening method and the 3.3 kb DNA fragment containing PEP gene was sequenced. DNA sequence from 20-2092 bp was ORF region coding 690 amino acids of prolyl endopeptidase. It was a new PEP gene through gene search. The genetic engineered E. coli BL21/pGEM-PEP overexpressing recombinant Aeromonas punctata prolyl endopeptidase(apPEP) was constructed. Cultured in YH medium, the expressed apPEP was about 30% of total cellular protein, the activity was 112 times more than that of wild strain. Expressed apPEP was mainly soluble intracellular protein, about 5% of apPEP was secreted to medium. Non-reduced SDS PAGE analysis showed it's monomer with molecular weight about 76 kD, which corresponded with prediction by gene sequence. Recombinant apPEP was purified after tube culture, the purity reached 90% and specific activity was 67 U/mg. PMID- 12548993 TI - [Study on production of poly-beta-hydroxybutyrate by recombinant strain VG1(pTU14)]. AB - Cloning and expression of Vitreoscilla hemoglobin gene (vgb) and lambda phage lytic genes (S-RRz) in production of Poly-beta-hydroxybutyrate(PHB) was studied in different Escherichia coli hosts such as E. coliJM105, E. coliJM109 and VG1. In the recombinant strains, VG1(pTU14) was a superior one which simultaneously contained three exogenes including vgb, S-RRz and PHB biosynthesis genes(phbCAB). The experimental results showed that after 82 hours fed-batch cult ure in LBG medium in shaking flask, cell concentration of VG1(pTU14) could reach 25.9 g/L, which was the highest PHB production ever reported, and till 52 h PHB content could be higher than 95%. Additionally, inducible cell lysis was also attained successfully in recombinant VG1(pTU14) accompanying with its high cell density culture. Therefore, VG1(pTU14) is a novel potential strain with promising prospect in industrial production of PHB. PMID- 12548994 TI - [Effects of nitrogen feeding on the accumulation of poly-beta-hydroxybutyrate with Alcaligenes eutrophus]. AB - On the basis of analysis of PHB fermentation processes, the effects of ammonium sulfate feeding rate at PHB formation period on the PHB accumulation by Alcaligenes eutrophus were investigated. It was shown that the complete absence of nitrogen source at PHB formation phase would lead to the decline of PHB synthetic activity, and the obvious influences of different nitrogen feeding rate on PHB synthesis were observed. Higher PHB content, but relative lower cell dry weight, PHB concentration and PHB productivity could be obtained at slower nitrogen feeding rate. The excessive nitrogen feeding rate resulted in the drop of PHB content, which led to the decrease of PHB concentration and PHB productivity. The better results could be achieved when the ammonium sulfate feeding rate was set at around 0.5 g/h. PMID- 12548995 TI - [Study on the improvement of plasmid stability in soybean rhizobia]. AB - By using luxAB as the report genes, 3.2 kb parCBA/DE gene fragment from pTR102 was inserted into pLAFR3 which contained a 3.7 kb enhancing fragment and deleted its cos site. Recombinant plasmids pHN155 and pHN156 were obtained. Contrasted plasmids pHN157 and pHN158 which contained cos site were also constructed. These four plasmids were transferred into Sinorhizobium fredii HN01 by tri-parental mating, and plasmid pHN155 and pHN158 were introduced into Bradyrhizobium japonicum TA11 by two-parental mating. The stability of the above plasmids was compared under free-living conditions and the results showed that the parCBA/DE could obviously enhance plasmid stability both in S. fredii and B. japonicum, and deletion of cos site showed only less effect. PMID- 12548996 TI - [Activities of superoxide dismutase in Xanthomonas oryzae pv. oryzae and its induction]. AB - The activities of superoxide dismutase(SOD) in Xanthomonas oryzae pv. oryzae were determined to demonstrate the correlation of SOD activities with the bacterial virulence. In liquid culture, SOD activities of the tested strains reached the maxima at the end of lag phase and then declined. The virulent strain PXO99A showed higher SOD activities than the avirulent strain PXO99A (pBUavr Xa10.F1). Analysis on the subcellular location indicated that SOD activities detected from the cytoplasm and periplasm were over 70% and 20%-30% of the total, respectively. Treatment of the bacterial culture for 1 h with exogenous O2- at concentration of 50-800 mumol/L induced SOD activities of both the strains. The treatment with 200 mumol/L resulted in the highest SOD activities. The effect of exogenous O2- on SOD induction was more significant in 12 h-culture than in 24 h-culture, and more notable for the virulent strain PXO99A. The treatment with exogenous O2- resulted in significantly decreased survival rate off both the strains. The decreases of survival rate were greater in 24 h-culture than in 12 h-culture, and more significantly for the virulent strain than for the avirulent one. PMID- 12548997 TI - [Purification and characterization of cuticle-degrading protease from entomopathogenic fungus, Metarhizium anisopliae]. AB - The cuticle-degrading proteases from entomopathogensis fungus, Melarhizium anisopliae, were induced by adding cicada exuviae, colloidal chitin, shrimp cuticle, maggot cuticle, horsefly cuticle and silkworm chrysalis cuticle into minimal medium. After ultrafiltration, Ultrogel AcA 54 column and IEF, a protease designated as MAP-21 with Mr 27 kD, and pI 7.6 were purified. It was shown that the recognition site of MAP-21 was Arg, PMSF and TLCK could inhibited the activity of this protease, indicating that there were Ser and His residues in the active center. The inhibitors to trypsin, leupeptin antipain and STI also repressed the activity of MAP-21, while chymostatin, TPCK and elastatinal TEI were shown no inhibition to its activity, demonstrating that, MAP-21 was a trypsin-like protease. Other properties of MAP-21 were also reported. PMID- 12548998 TI - [Effects of soil factors on root colonization of wheat by luxAB genes-marked Pseudomonas fluorescens Xl6L2]. AB - Colonization density of Pseudomonas fluorescens Xl6L2 marked with luxAB genes in wheat Rhizosphere in sterilized Rhizobox--Calcareous chao soil microcosms was greater than that in insterilized microcosms. The population of Pf.Xl6L2 in the Rhizosphere in the rhizobox--Yellow brown soil microcosms was larger than that in the Calcareous chao soil microcosms; As soil water content was about 50% field capacity (FC), Pf.Xl6L2 could move to the place of 8 cm of root from coated seeds in the former microcosm, but only 4 cm in the later microcosm. In the former microcosms, the number of Pf.Xl612 in the rhizosphere under 60% FC and 75% FC were greater than that under 50% FC; In the second microcosm, the dispersal distance of Pf.Xl6L2 along the root was affected significantly by the soil moisture, it could be detected as far as 8 cm of root from the coated seeds under 60% FC and 75% FC. PMID- 12548999 TI - [Study on fermentation of 1,13-tridecanedioic acid by Candida tropicalis]. AB - A mutant of Candida tropicalis SP-1, SP-UV-56, which can produce 1,13 tridecanedioic acid or 1,11-dicarboxylic acid (DCA13) 1.25 times as much as its parental strain SP-1, but hardly assimilates nalkane as carbon source for its growth, was obtained by ultraviolet treatments. A fermentation technology with a supplement of acetate at the latter middle of logarithmic growth was developed. Using sucrose as carbon source for its growth and supplying acetate the yield of DCA13 reached 153 g/L for 144 h incubation in a 13.7 L auto-controlled stirred tank, about 29.7% higher than that obtained in the broth without acetate. An effective technique that can maintain higher yield was provided by enhancing mixing of broth and controlling low DO level. With these techniques the DCA13 yield reached 172 g/L in a 20 m3 stirred tank, and produced 2.25 tons DCA13 in 15.0 m3 broth. PMID- 12549000 TI - [Cloning and expression of the gene encoding novel alpha-amylase from Sulfolobus shibatae in E. coli]. AB - A novel alpha-amylase gene was amplified from Sulfolobus shibatae by using PCR technique. The amplified 1.7 kb DNA fragment was inserted into an expression vector pBV220 to yield the recombinant plasmid pSBAM. The novel alpha-amylase gene in pSBAM was expressed in E. coli. The production of the novel alpha-amylase activity reached over 8 units/100 mL of the culture. The molecular weight of this enzyme was about 61 kD by SDS-PAGE. The expressed novel alpha-amylase protein in E. coli DH5 alpha accounted for about 20% of the total protein in the recombinant cell. The cooperative action of the novel alpha-amylase and the maltooligosyltrehalose synthase from Sulfolobus shibatae was investigated and trehalose was detected by using HPLC analysis when using amylose and partial starch hydrolysates as substrates. PMID- 12549001 TI - [Biomachining of metal copper by Thiobacillus ferrooxidans]. AB - Thiobacillus ferrooxidans was employed in the biomachining process of metal copper(Cu0). The bacteria growth and the changes of Fe3+ concentration during machining processes have been studied. Biomachining and chemical machining have been compared. The results showed that the concentrations of bacteria and Fe3+ determine the speed of machining copper. The biomachining is more fast that chemical maching because bacteria are able to regenerate Fe3+ oxidizing copper. It was also found that the Cu2+ produced from the machining processes inhibit the growth of bacteria. Cu2+ has to be removed. PMID- 12549002 TI - [Microbial genomic research and its impact on biological science]. PMID- 12549003 TI - [Transposable elements in Streptomyces spp]. PMID- 12549004 TI - [1.4 kb 5' flanking region of class I patatin directs tuber-specific gus expression in potato (Solanum tuberosum L.)]. AB - Binary vectors pPATIs (with partial signal sequence) and pPATI (without signal sequence) were constructed by fusing 1.4 kb 5' flanking regions of Class I patatin gene with GUS. Transient GUS expression was observed in in vitro tuber slices bombarded with pPATI. These constructs were then introduced into potato (cv. Desiree) via Agrobacterium tumefaciens transformation. Transgenic potato plants were confirmed by X-Gluc staining and PCR. Using in vitro tuberization system, GUS activities were assayed by fluorescence. It was shown that, in plants transformed with PATI-GUS, GUS specific activities were about 10-20 fold higher in tubers than in stems. Increased sucrose concentration could not induce PATI GUS expression, but light enhanced PATI-GUS expression in cultured shoots. PMID- 12549005 TI - [Selection and identification of salt tolerant line of sainfoin from the seeds of first post-flight plants]. AB - Seeds of sainfoin (Onobrychis vicaefolia Scop.) were carried in the recoverable satellite 940703 and recovered from earth orbit from China in 1994. The progeny seeds were obtained by producing in field. The salt tolerant calluses were selected by screening seedling and callus on 1.5% NaCl-containing medium, reviving growth on NaCl- free medium and selecting callus on 1.2% NaCl-containing medium. The salt tolerant line callus maintained the normal ability to regenerate plant. The salt tolerant line callus exhibited cross-resistance to PEG stress. The variant appeared higher efficiency than control to accumulate proline under salt stress, however, under nonstress condition it had lower proline level than control, which suggested that the higher efficiency to synthesize proline under stress condition may be more important than higher level in tissue under nonstress condition. The mechanism of proline synthesis in the selected callus was considered to result from the alteration in gene sensitivity to water regulation at transcription level. Acrylamide gradient electrophoresis showed that new isoenzyme form with MW 175 kD and 75 kD of superoxide dismutase and esterase respectively appeared in salt torlerant callus. It was indicated that the combination of space mutagenesis with tissue culture could be used for the selection of salt tolerant sainfoin line in vitro. PMID- 12549006 TI - [Immunogold localization of ribulose-1,5-bisphosphate carborylsae/oxygenase in chloroplasts of Chlorella]. AB - Ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a first key enzyme in the Calvin Circle of plant cell photosynthesis. This paper mainly studied gold immunolocalization of Rubisco of Chlorella spp. 640909, and the Native-PAGE and, SDS-PAGE and Western bloting analysis, as well as the observation to pyrenoid ultra structure. The Native-PAGE result showed a main band, evidenced as the Rubisco band by the Western blot with the antibody against the Rubisco from C. prototothecoides, The special immunoacton of Rubisco from Chlorella spp. 640909 and the antibody to large subunit of Rubisco from C. prothecoides showed the large subunit proteins of Rubisco in the two species of Chlorella shared the high homology. The SDS-PAGE and Western blotting maps showed the molecule weight of the large subunit of Rubisco of Chlorella spp. 640909 was about 55 KD. The shape of pyrenoid ultra structure of the electronic microscope was oblong, and was embedded in starch sheath, with 2 swelling thylakoids through out a center portrait channel of the pyrenoid. There were some connections between pyrenoid and the chloroplast stroma. The distribution of the large subunits and the whole Rubisco in the chloroplast of Chrolella spp. 640909 was studied by immunoelectron microscopy by embedded sections with antibody to large subunit and whole enzyme followed by second antibody, goad anti-rabbit immunoglobulin G conjugated to 10 nm gold particles(Sigma production). The result showed the antibodies against large subunit and whole enzyme heavily labeled the pyrenoid, as well as starch sheath region, whereas the thylakoid region of the plastid was lightly labeled. And the whole Rubisco antibody labeled the pyrenoid surface more heavily than the large subunit antibody did. It is demonstrated the pyrenoid and starch sheath have the photosynthesis function. Rubisco concentrating in pyrenoid and starch sheath is valuable to fix CO2 for photosynthesis in algae. PMID- 12549007 TI - [Knocking out of CEK2 and CSK1 affect the phenotype of Candida albicans]. AB - We have cloned two novel mitogen-activated protein kinase (MAPK) genes CEK2 and CSK1 from Candida albicans strain SC5314. The heterozygous and homozygous cek 2 and csk 1 mutants were obtained by homologous recombination. The cek2 and csk1 null mutant strains CAJ3 and CAJ6 grew as wrinkled clonis on solid YPD medium. Expression of CEK2 gene in strain CAF3-1 repressed the filamentous growth of the strain weakly. Two-hybrid results showed Cek2 and Csk1 could not interact with Cph1 and Hst7 directly. Cek2 and Csk1 might cooperate with the filamentous transcriptional repressor Tup1. PMID- 12549008 TI - [The mechanism of neuro-2a cell death induced by H2O2]. AB - Reactive oxygen species (ROS), such as H2O2, can be produced by enzymes involved in electron leakage of respiration chain in mitochondria, and by neurochemical enzymes such as monoamine oxidase in neural cells. ROS are toxic to cells, and can result in cell death. ROS also play an important role in some diseases, especially in neurodegenerative diseases by yet unknown mechanisms. In the current research, the N-2a neuroblastoma cell was treated with H2O2, and the morphological changes of cell death were characterized. Our results show that N 2a cell death is different from classical apoptosis, but belongs type II nerve cell programmed death, which shows condensed chromatin within intact nuclear envelope and no apoptotic body. The chromatin DNA of dead cells shows no internucleosomal cleavage, as well as no requirement for caspase-3, 1 activity. However, the H2O2-induced N-2a cell death can be inhibited by Bcl-XL. It can be concluded that type II nerve cell death is the result of cell toxicity mediated by ROS. The results pave the way for further research of type II nerve cell death. PMID- 12549009 TI - [A study on the development of the nitrergic neurons in the large intestine of human fetus]. AB - By using histochemical methed of NADPH-diaphorse, the development of the nitrergic neurons in the large intestine of human fetus were studied. The results showed: At the fifth month of gestation, weak positive reaction of nitric oxide synthase (NOS) appeared in part of the round cells of intermuscular ganglia. The round cells differentiated into the nitrergic nerve cells. At the sixth month, the bodies of nitrergic neurons were obviously enlarged, the processes of which were lengthened. The nitrergic nerve fibers were seen in the muscle layer, the submucosa and the base of the intestinal gland. The growth and development of nitrergic neurons reached its peak at the seventh month. The number of intermuscular ganglionic cells was increased. The density of nitrergic nerve fibers was increased in the inner circular muscle layer, and have bead-like structures. At the eighth to tenth month, the staining intensity of nitrergic neurons was increased. The myenteric plexus was densely distributed with nitrergic nerve cell bodies, whereas the submucosa and the inner circular muscle layers contained only a few neurons. The nitrergic nerve fibers were observed in all layer of large intestine, the density of the distribution of nitrergic nerve fibers was by far the highest in the inner circular muscle layer, less in the submucosa and outer longitudinal muscle layer, and only a few were found in the mucous layer. To our knowledge, it is the first time that the development of nitrergic neurons in the large intestine of human fetus was demonstrated. PMID- 12549010 TI - [Higher lint percent and drought tolerant cotton line selected by radiation breeding]. AB - The Gossypium hirsutum cv. Liaomian No. 9 were mutagenized by 60Co gamma ray, from which the mutant line Zhonghuzhi PI 935 (be called "PI 935" for short) was bred by family selection method. The PI 935 not only has some good traits (growing period, drought tolerance, lint color and fiber quality) similar to the original cultivar, but also has higher lint outturn and lint yield than that of the Liaomian No. 9. The PI 935 has been identified and regional tested in nine places times for four years in the southern Xinjiang Weiwuer autonomous region. It was shown that the PI 935 had the higher lint outtrn for the average 47.3% was ten-point percentage more than that of the check cultivars (Junmian No. 1 or Xinluzhong No. 5), the similar lint yield by and large and the growing period by five days later than that of the checks. The PI 935 was collected in the National Bank of Crop Germplasm (unified No. ZM 114274 and named "Zhonghuzhi PI 935"). PMID- 12549011 TI - BRCA1 is differentially expressed in human tumor cells. AB - Mutations of the human breast cancer susceptibility gene 1 (BRCA1) confers a risk for breast, ovarian and prostate cancers and BRCA1 exerts multiple biological functions. Using Western blot and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) assays, we have determined the expression of endogenous BRCA1 protein and mRNA in forty-three human tumor cell lines established from eleven types of human tumor tissues. BRCA1 was differentially expressed in tumor cell lines. No significant association was found between BRCA1 expression and the p53 gene status of cell lines. The disruption of wild-type p53 by either the human papillomavirus E6 oncogene or the mutant p53 gene (143Ala- >Val) did not cause any significant alteration in basal level of BRCA1 expression, while the knockout of p21 (-/-) by homologous recombination assay and Blocking Gadd45 expression by constitutive antisense expression slightly increased BRCA1 protein expression. Therefore, although the functional significance of the differential expression in human tumor cells is currently unknown, the present data provide a valuable background for further study of BRCA1 in tumor cell lines. PMID- 12549012 TI - [Effect of alpha-lipoic acid on the apoptosis of PC12 cells induced by 6 hydroxydopamine]. AB - PC12 cell line, a clonal cell line derived from a pheochromocytoma of rat adrenal medulla, was used as a model of dopaminergic neuron in vitro to study the effect of alpha-lipoic acid on the 6-OHDA induced apoptosis. The results from MTT method show that 6-OHDA decreased the cell survival rate obviously. Through TUNEL (TdT mediated dUTP-biotion nick end labeling) and Flow cytometer (FCM) detection, we found that 6-OHDA triggered cell apoptosis and induced necrosis. It was confirmed by the different percentage of cell survival rate and apoptosis concluded from FCM and MTT. alpha-lipoic acid was used as antioxidant to protect the cell from 6 OHDA's injury. The result indicateed that alpha-lipoic acid can partly prevent apoptosis induced by 6-OHDA but fail to prevent necrosis since it can decrease the apoptotic cell from 20.09% to 3.09%, just as increased cell survival rate from 56.8% to 72.6% but can not reach the normal level showed by MTT assay. Biochemical approach showed the cell's antioxidant ability especial for SOD activity and GSH content increased after the treatment of alpha-lipoic acid. The data suggest that alpha-lipoic acid may protect PC12 cells from apoptosis induced by 6-OHDA through the antioxidant path. PMID- 12549013 TI - [Studies on ABA-induced H2O2 in Vicia guard cells by means of confocal laser scanning microscopy]. AB - The methods of confocal laser scanning microscopy (CLSM) and microinjection were used to study ABA-induced H2O2 in guard cells (Vicia faba), which were labeled with H2O2 specific probe-2, 7-dichlorofluorescin diacetate(H2DCFDA). The results indicated 100 U/mL catalase (CAT) could inhibit partly stomatal closure induced by ABA. 10(-3) mmol/L ABA could significantly induce H2O2 production in chloroplast in guard cells of Vicia faba following microinjection, and 100 U/mL CAT could partly abolish the effects following simultaneous microinjection of ABA and CAT. These suggest that H2O2 is possibly involved in ABA signaling leading to stomatal closure. PMID- 12549014 TI - [The effect of autocrine factors on development of early embryos of mouse]. AB - Two-cell-stage embryos were flushed from the oviducts on Day 2. Zygotes were collected from oviducts on Day 1 (Fertilization In Situ, ISF) or derived from fertilization in vitro (IVF). 2-cell embryos had a high rate of blastocyst development to each embryo concentration from 1 embryo/microliter to 1 embryo/1000 microliters. The zygotes produced by either ISF or IVF were adversely affected by reducing the embryo concentration over this range (P < 0.001), with approximately 82.5% of ISF zygotes developing to blastocysts at highest concentration but only 22.3% at the lowest. For IVF zygotes the corresponding results were 46.3% and 5.2%. The number of cells in each blastocyst from 2-cell embryos was significantly higher than that from ISF and IVF group. The media supplementing Platelet-activating factor (PAF) caused a significant increase in the rate of blastocyst development of IVF zygotes at embryo concentration of 1 embryo/10 microliters (10 ng/ml) and 1 embryo/100 microliters (100 ng/ml). Insulin-like growth factor (IGF) (10 ng/ml) also stimulated development of IVF zygotes when they were cultured at the concentration of 1 embryo/10 microliters. Epidermal growth factor (EGF) was no effect over range of 1-1000 ng/ml to embryo development. The results show that factors necessary for normal embryo development are diluted to suboptimal levels during culture at low embryo concentration. The PAF, IGF-I partially compensate the effects of low embryo concentration during culture and play important roles as autocrine embryotrophic factors. PMID- 12549015 TI - [A sinorhizobium fredll strain that effectively nodulate Medicago sativa]. AB - Sinorhizobium fredii 042BS was isolated from root nodules of alfalfa (Medicago sativa) from Xinjiang Region. Nodulation experiments showed that both soybean and alfalfa were nodulated by 042BS effectively. The 16S rDNA PCR-RFLP analysis was carried out by four restriction endonucleases, and the restriction maps of strain 042BS were identical with those of S. fredii USDA205. The DNA G + C mol% of strain 042BS was 60.0. The DNA homology between 042BS and S. fredii USDA205 and USDA191 were 84.9% and 89.6%, respectively. To prove the capability of 042BS to nodulate both soybean and alfalfa, constitutively expressed green fluorescence protein gene(gfp) was introduced to 042BS, and the recombinant strain 042BSG was obtained. The reisolates from nodules of the soybean and alfalfa inoculated with 042BSG were observed using the confocal laser-scanning microscope, and the expressions of gfp were detected, respectively. 042BS showed various nodulation capacities with different alfalfa cultivars used. PMID- 12549017 TI - [Cloning of parasporal body protein gene resembling to S-layer protein genes from Bacillus thuringiensis CTC strain]. AB - Bacillus CTC strain was identified as Bacillus thuringiensis subsp. finitimus (serotype H2) and Pasteur Institute confirmed this identification. The parasporal body formed by CTC strain is oval shaped, and consists of 100 kD protein. The determination of the N-terminal amino acid sequence showed this protein shares 93% similarity to that of B. anthracis S-layer proteins. The restriction map covering the related protein gene (ctc) was deduced according to southern hybridization. The DNA fragments containing the 5' and 3' end of ctc gene with 0.6 kb overlap were isolated, respectively. Full length ctc gene was then constructed. B. thuringiensis is a species similar to B. cereus and B. anthracis, and only can be distinguished by forming parasporal bodies. The growth of E. coli maintaining ctc gene is similar to that of E. coli expressing S-layer protein gene. The above preliminary results showed that the parasporal body protein in CTC strain is a cell surface-like protein. So identifying CTC strain as a species of B. thuringiensis and the unique criterion for the identification of B. thuringiensis by forming parasporal body should be reconsidered. PMID- 12549016 TI - [Extraction of DNA from environmental samples and construction of mixed genomic DNA libraries]. AB - A method has been developed for extracting and purifying genomic DNA from environmental samples. In this method, an environmental sample is treated first by grinding and freezing/thawing and subsequently by SDS/proteinase K-based DNA extraction. The yields of purified DNA from three samples used in this study ranged from 2 to 16 micrograms per gram of dry sample. Mixed genomic DNA libraries for two of the environmental samples were constructed by inserting restriction fragments (3-8 kb) of the purified DNAs into plasmid pUC18 and transforming E. coli DH5 alpha with the resultant plasmids. Approximately 10(3) to 10(4) insert-containing clones were obtained from 1 g of each sample. Clone libraries were analyzed by DNA sequencing and gene annotation. Among 20 randomly selected clones, 14 contained an insert whose sequence had not been reported while the rest had an insert of either E. coli or vector origin. A search of sequence databases using the end sequences of each of the foreign inserts showed that each sequence was part of a gene encoding, in most cases, a predictable function. Our results are of significance to the collection, investigation and exploitation of the genes of uncultured microorganisms. PMID- 12549018 TI - [Detection of enterotoxin genes in Bacillus thuringiensis by PCR]. AB - The presence of hemolysin HblA (hblA), enterotoxin BceT (bceT) and enterotoxin S (entS) genes from 45 strains of B. thuringiensis, 4 strains of B. cereus and B. sphaericus have been detected respectively by multiple PCR. The results showed that 95.6% B. thuringiensis strains contain the B component of hblA gene, 91.1% and 93.3% of them contain bceT and entS genes sequences respectively. The enterotoxin productions in all strains have also been analysis using two commercial immunoassay kits(TECRA and RPLA) and it has proved that most of B. thuringiensis stains and the positive B. cereus strain can produce entero toxins during their growth. However, the two hblA sequence positive stains, DBT007 was negative when tested both by RPLA and Tecra, T24 001 was negative when assayed by Tecra and positive by RPLA. One hblA sequence negative strain Dmu39 was negative when tested by RPLA but positive by Tecra. No enterotoxin and enterotoxin gene could be detected in B. thuringiensis DBT248 and the B. sphaericus strains. The results suggest that the potential risk of using B. thuringiensis, as biopesticide needs to be further evaluated. PMID- 12549019 TI - [The clone and comparison of very virulent plus Marek's disease virus(vv+ DMV)648 strain glycoprotein E(gE) gene]. AB - The glycoprotein E(gE) gene of very virulent plus Marek's Disease (vv+ MDV) 648 strain was amplified by polymerase chain reaction (PCR) and cloned into pUC18 vector. The gE sequence was determined and analysed. The whole length of 648 strain gE gene is 1494 base pairs. The protein encoded by gE gene has several features characteristic of a membrane-associated glycoprotein. It contains eight potential glycosylation sites, a markedly hydrophobic region at the N terminus that could function as a signal peptide and a hydrophobic segment (aa 391-419) at C terminus that could function as a transmembrane anchor element. Comparison of the gE between 648 strain and virulent MDV (vMDV) GA strain revealed that gE sequence is conserved in MDV serotype 1, there are only two basepairs different, (at bp 512 and 1472), which induce two amino acids changed respectively (at aa 171 and 491). The function of 648 strain gE and the difference of gE between 648 strain and GA strain is on study. PMID- 12549020 TI - [Expression of rice dwarf virus outer coat protein gene(S8) in insect cells]. AB - Outer coat protein gene(S8) of RDV was cloned into the transfer vector pVL 1393 to construct a recombinant vector pVL1393-S8. The recombinant vector pVL1393-S8 and the linear baculovirus RP23. LacZ were cotransfected into sf9 cells to produce the recombinant virus RP23-S8. RP23-S8 infected sf9 cells were collected and analysed by SDS-PAGE and Western-blot. The results showed that the S8 gene of RDV was expressed in sf9 cells and the expression level of sf9 cells was higher between 72-96 h after infected. PMID- 12549021 TI - [Expression of two truncated enhancin gene from Helicoverpa armigera granulosis virus in E. coli and its preliminary bioassay]. AB - The plasmids pET-30a-Ben and pET-30a-Den which included 1.7 kd and 2.2 kb fragments of 5'-terminal of HaGV enhancin gene were obtained by cutting recombinant plasmid pET-30a-En with Bal I and Dra I respectively. Two fragments were expressed in E. coli successfully and the products were named Ben and Den respectively. The enhancement, which Ben and Den enhance the infectivity of HaNPV and Bt in 3rd larvae of Helicoverpa armigera, was studied. The results indicated that there was increase of the mortality of 10.5%-26.5% and the LT50 decrease of 0.9 d causes by adding Ben, while Den could increase the mortality by 10.2%-33.0% and decrease the LT50 by 1.2 d-1.9 d. The preliminary bioassay on Bt against Helicoverpa armigera indicated the recombinant enhancin could increase the mortality of larvae by 20.7%-35.4%, Den by 16.7%-31.5%, Ben by 11.7%-27.4%. PMID- 12549022 TI - [Analysis and expression of Hyphantria cunea nuclear polyhedrosis virus sod gene]. AB - The sequencing results indicated that Hyphantria cunea nuclear polyhedrosis virus (HcNPV) sod gene open reading frame of 456 nt encoding protein of 151 amino acid, was identified to that of Bombyx mori nuclear polyhedrosis virus (BmNPV), and exhibited 97.2% homology at nucletde level to that of Autographa californica nuclear polyhedrosis virus (AcNPV), three amino acid residues difference in amino acid level with AcNPV sod. The essential amino acid residues for the construction and active could be detected in HcNPV sod. Activity of the SOD is 147.09 U per milliliter E. coli. PMID- 12549023 TI - [Cloning of promelittin cDNA and its expression in Escherichia coli]. AB - The cDNA encoding promelittin was obtained from the total RNA of bee poison gland by RT-PCR and was cloned to pT7Blu-T vector. The expression vector of promelittin fused with partial sequence of beta-galactosidase was constructed by ligating the fragment inserted to pUC118. Moreover, it was expressed in the strain DH5 alpha of Escherichia coli. The result of DNA sequence analysis demonstrated that the obtained cDNA sequence was same with the published one and the reading frame of fusion gene was correct. PMID- 12549024 TI - [Purification and gene cloning of a novel fibrinolytic protease from Streptomyces sp. C3662]. AB - A novel fibrinolytic protease from Streptomyces sp. C3662 was purified by (NH4)2SO4 fractionation, DEAE-Sepharose and CM-Sepharose chromatography. The molecular weight of the protease was indicated to be 30 kD by SDS-PAGE. Using a E. coli/Streptomyces shuttle plasmid pIJ699 as the vector, shot-gun cloning was performed to clone the gene of the protease. One clone with fibrinolytic activity harboring a plasmid that contains a DNA fragment of 6 kb was obtained from 3000 clones. Sequence analysis reveals that the open reading frame of the gene of the protease is 903 bp in size, encoding a putative protein of 300 amino acids with 30 kD. The overall GC% and the third codon GC% of the ORF were 68.33% and 95.6%, respectively. Comparison of homologue showed that the purative protein is highly homologous with other proteases of Streptomyces sp. PMID- 12549025 TI - [Biosynthesis of glutathione: construction of ATP regeneration system between recombinant E. coli and S. cerevisiae]. AB - An ATP regeneration system between two microorganisms, in which recombinant E. coli II-1 and S. cerevisiae WSH-J702 were used as the biosynthesis enzymes source of glutathione and ATP, respectively, was constructed by using glucose as an energy substrate. Yeast cells after permeable treatment could hardly consume glucose. The glycolysis pathway of yeast was started by adding 1.0 mmol/L AMP and 0.05 mmol/L NADH simultaneously. GSH biosynthesis in the coupling system was improved with increasing glucose concentration to 400 mmol/L, where the concentration of GSH reached 10.4 mmol/L (3.2 g/L). Both the coupling system and the non-coupling system with ATP added could hardly produce GSH under the absence of magnesium. The low efficiency of ATP regeneration system constructed was probably caused by the chelation between magnesium and ATP. By adding magnesium to the coupling system, a final concentration of GSH, 14.3 mmol/L (4.4 g/L), was achieved at 6 h of reaction. PMID- 12549026 TI - [Purification and properties of neutral phytase form Bacillus subtilis]. AB - A strain Bacillus subtilis producing neutral phytase was screened from soil. The protein of phytase was purified by HPLC. Optimal pH value and temperature of the phytase for its activity were 7.5 and 55 degrees C, respectively. The Km values of the phytase for dodecasodium phytate under 37 degrees C was 0.19 mmol/L. The molecule weight of the phytase protein was determined as about 45 kD by SDS-PAGE. The N-terminal amino acids sequence of the phytase protein was determined as Lys His-Lys-Leu-Ser-Asp-Pro-Tyr-His-Phe-Thr by amino acids sequence analysis. PMID- 12549027 TI - [Studies on the immobilized penicillin acylase on polymer beads]. AB - The extracellular penicillin acylase from Bacillus megaterium was immobilized on oxirane group of Eupergit C beads. The apparent activity of the immobilized enzyme was about 1400 u.g-1 (dry weight). The optimal pH and temperature were 8.0 and 55 degrees C for hydrolytic reaction of penicillin G, respectively. The immobilized enzyme was stable in the pH range of 6.0-8.5 and at temperature below 45 degrees C. The apparent Michaelis constant for penicillin G was inhibition constant of phenylacetic acid as competitive 2 x 10(-2) mol.L-1 and Vm was 1.33 mmol.g-1 min-1 (dry weight) at 37 degrees C and PH8.0. The inhibitor and 6-APA as non-competitive inhibitor were 2.8 x 10(-2) mol.L-1 and 0.125 mol.L-1 for the immobilized enzyme at pH 8.0 and 37 degrees C, respectively. The remained activity of the immobilized enzyme was about 80% after operating 200 times for hydrolysis of penicillin G to 6-APA, and the average yield of 6-APA was 89.48%. PMID- 12549028 TI - [Studies on the methods of mating test for Cryptococcus neoformans]. AB - Six media (MA:Mating agar, HA:Hay agar, HCA:Hay cube agar, RSA:Rice shell agar, IHA:additional nutrition in HA and SDA:Sabouraudia dextrose agar) were compared for their efficiencies of the mating test for Cryptococcus neoformans. The positive rates in 2-week-cultivation were 95% (HCA and RSA), 86% (HA), 38% (MA and SDA) and 24% (IHA). The streaking inoculation was equally replaced by the dot inoculation for the mating tests. However, the dot method saves much more materials, and is suitable for testing in large number of isolates. Additionally, the precultivation of the tester and tested isolates in broth medium (YMPDB) could enhance the positive mating rates. PMID- 12549029 TI - [Isolation, purification and identification of metallothionein from strain BD102 of Hansenula anomala]. AB - Metallothionein (MTs) in Cu and Cd resistant strain BD102 of Hansenula anomala were induced by administration of Cu2+ and Cd2+. These proteins were isolated and purified by Sephadex G-50 and subsequent DEAE-Sepharose CL-6B, then Sephadex G-25 for desalation. There were two isoform MTs by Cu(Cu-MTs), one form induced by Cd (Cd-MT). The molecular weights of the Cu-MTs and the Cd-MT were about 7 kD and 7.5 kD respectively. Exposure of Hansenula anomala to copper salts stimulated formation of two isoform Cu-MTs with a cysteine content of 6.6-6.8% and had 60 amino acids. Exposure of Hansenula anomala to cadmium, stimulated formation of Cd MT with a cystein content of 10% was synthesized and had 61 amino acids. 4 atom Cu or Cd/mole MTs. PMID- 12549030 TI - [Screening and characteristics of mutants of E. coli resistant to acetate inhibition]. AB - Acetate is accumulated in the aerobic high cell density culture of Escherichia coli, Which seriously inhibits cell growth and expression of recombinant proteins. To alleviate acetate inhibition, mutants of E. coli JM101 were generated by 60Co radiation and then enriched in continuous culture with acetate as the selective pressure. One of the mutants isolated, JL3, showed obvious increased tolerance toward acetate and maintained phenotypic stability on slant without acetate. In MA medium containing 10 g/L of sodium acetate, the specific growth rate and the glucose consumption rate of JL3 were higher than those of JM101. PMID- 12549031 TI - [A study on the breeding of new Ganoderma varieties by UV induced mutagenesis]. AB - UV induced mutagenesis of Ganoderma's protoplast was carried out. By crude screen and careful screen, we obtained two mutants, 43020# and 43026$, whose dry weight and polysaccharides contents are higher than original strain. Results of continued cultivation for 10 generation on PDA slants and successive tests of shaking bottles, and successive 3 time pilot scale tests at 3 tons, showed that these two mutants are excellent strains with steady properties, high production and high content of polysaccharides. This study has provided a rapid and effective method for breeding of Ganoderma varieties which is suitable to industrial fermentation. PMID- 12549032 TI - [Effect of various substrate nutrient on the spore germination and infective germ tube inducing of Pandora delphacis]. AB - The influence of nutrients on spore germmination of P. delphacis was investigated using amino acids, fatty acids, carbohydrates, vitamins, minerals, and extracts of aphids or planthoppers. It was found that the formation of germ tubes from primary conidia was significantly stimulated by cysteine, asparagine, behenic acid (C22:0), trehalose, fructose, glycerol, maltose, ascorbic acid, thiamine(VB1), Folic acid, ZnSO4, FeSO4 and extracts of aphids and planthoppers. Of these many could suppress the formation of secondary conidia from primary conidia, e.g., cysteine, behenic acid (C22:0), myristic acid (C14:0), palmitic acid (C16:0), stearic acid (C18:0), arachidic acid (C20:0), lignoceric acid (C24:0), ascorbic acid, thiamine (VB1), riboflavin (VB2), folic acid, nicotinic acid, ZnSO4, and FeSO4. However, the nutrients not conducive to the formation of germ tubes were stimulants to the formation of secondary conidia. These included tyrosine, alanine, glucose, galactose, sucrose, glycogen, inositol, pyridoxal (VB6), NaCl, and KCl. Moreover, nutrients including asparagine, trehalose, glycerol, maltose, fructose, thiamine (VB1), and folic acid stimulated the formation of both germ tubes and secondary conidia. The result above indicated that the substrate nutrients which can stimulate the formation of germ tubes were general in planthoppers and aphids, it means the formation of aggressive germ tubes was one of important factors of successful for infection to the hosts. Of which it confirmed that the host specificity for P. delphacis against N. lungus and M. persicae. PMID- 12549033 TI - [Isolation of a development revertant of Myxococus Xamthus]. AB - fruA::Tc omega 5 is a development deficient strain of M. Xamthus. Transposon TnV was used to randomly mutagenize various sites of fruA::Tc omega 5 chromosome. Fruiting body formation was restored in one TnV insertion mutant, designated XM1206. The TnV-inserted DNA fragment from XM1206 chromosome was cloned, which may be served as a probe to isolate the corresponding allele from wild-type strain. PMID- 12549034 TI - [High level expression of human recombinant albumin gene in the yeast Pichia pastoris]. AB - The yeast Pichia pastoris was transformed by the multi-copy Pichia expression vector that can express secreted human albumin. The high level expression of cell line was selected after screening. The expression of human recombinant albumin in Pichia pastoris induced by different methods were compared. The ratio of secreted human albumin is 80% in total secreted proteins and the expression level reaches as high as is 10 g/L. PMID- 12549035 TI - [Cloning and expression of D-arabitol dehydrogenase gene from Acetobacter suboxydans in Escherichia coli]. AB - The partial genomic library of Acetobacter suboxydans was constructed using Yeast E. coli shuttle plasmid YEp352 as vector. Two positive transformants, designated as DH5 alpha(pAD91) and DH5 alpha(pAD98), were obtained by screening the growth of transformants on the agar plate in which D-arabitol was used as the sole carbon source. The results of Southern blot and restriction endonuclease analysis showed that the two recombinants are identical. The insert is about 2.3 kb. Arabitol dehydrogenase activity assay indicated that the transformants could produce D-xylulose-forming D-arabitol dehydrogenase. Hence, the gene encoding D arabitol dehydrogenase exists in the cloned DNA fragment. PMID- 12549036 TI - [Post-genome studies on Mycobacterium tuberculosis and novel anti-TB vaccines]. PMID- 12549037 TI - [Cloning of cancer-related genes of Rat6 fibroblasts by using an improved differential display method]. AB - The p53 gene is the most frequently mutated gene identified so far in human cancers. When a mutant p53(135)-val gene was allowed to be over-expressed in Rat6(R6) cells, a high incidence of spontaneous transformation was observed in long-term culture of this cell line(R6 # 13-8). To identify genes involved in cell transformation, parental p53 over-expressing cell, R6 # 13-8, and its spontaneous transformant T2, were analyzed by an improved mRNA differential display technique, which was reproducible, simpler, and was able to clone cDNA longer than 500 bp, and was with less false positives. When 33 10-mer or 12-mer single primers with arbitrary but defined sequence were used for PCR, over 90 discrete cDNAs were obtained from R6 # 13-8 and T2 cells. Three differentially expressed cDNAs were identified, one of them is highly expressed in T2 cells, while the other two, 0.8 kb and 0.9 kb long, are highly expressed in R6 # 13-8 cells. The latter were cloned and confirmed by Northern hybridization. Both cloned fragment were not homologous with any published sequence. Our results suggest that the activation and inactivation of genes are involved in the process of the spontaneous transformation from R6 # 13-8 to T2. PMID- 12549038 TI - [Cloning and tissue expression pattern analysis of the human citrate synthase cDNA]. AB - Tricarboxylic acid (TCA) cycle is an important way to generate ATP, which is widely distributed in the cells of animal, plant or microorganism. It catalyses the catabolism of sugar as well as protein and fat. Citrate synthase plays a key role in regulating TCA cycle and is responsible for catalysing the synthesis of citrate from oxaloacetate and acetyl CoA. Screening of genomic informatics was performed by using pig citrate synthase cDNA as a probe and a contig which is 1636 bp long and has highly homologous to the pig citrate synthase cDNA was obtained from selected ESTs with the ASSEMBLY program. According to the sequence of this contig, a pair of primers was designed and used to amplify cDNA libraries. A 1492 bp cDNA containing an open reading frame encoding 466 amino acids was cloned from human testis and skeletal muscle cDNA libraries. The deduced amino acid sequence of the cDNA showed 95%, 92% and 60.9% identity to pig, chicken and yeast citrate synthase respectively. Because the deduced amino acids sequence contains a highly conserved motif of citrate synthase from three different species, it is believed that this cDNA may be a transcript of human citrate synthase gene. Northern analysis showed that the human citrate synthase was expressed at high level in heart and muscle, at middle level in brain, kidney and pancreas tissues, not detectable in thymus and small intestine tissues, and at low level in other nine tested human tissues. PMID- 12549039 TI - [Effects of hypotonic stress on the survival of hatched mouse blastocysts pericryopreservation]. AB - The objective of this study to evaluate the effect of hypotonic stress on developmental potential of hatched blastocysts perivitrification. Hatched mouse blastocysts were vitrified in liquid nitrogen after equilibration in 10% or 20% GL for 5 min and in GFS40 for 30 sec respectively, the survival rates were 93% 97% after the frozen-thawed embryos were cultured in vitro for 24 h. There were no statistical difference between the frozen and the fresh group (P > 0.05). In order to evaluate effects of hypotonic stress on developmental abilities, fresh hatched mouse blastocysts were respectively exposed to 1.00 x, 0.50 x, 0.30 x, 0.25 x and 0.20 x PBS for 30 min, then cultured in mKRB for 24 h, the survival rates were 98%, 99%, 92%, 92% and 50% respectively. The rate in 0.20 X PBS group was significantly lower than in other groups (P < 0.01). When frozen-thawed embryos were directly treated with different osmotic solutions, the survival rates were 88%, 72%, 58%, 11% and 0 respectively in 1.00 x, 0.50 x, 0.30 x, 0.25 x and 0.20 x PBS group. The rate in 1.00 x PBS group was significantly higher than in other groups (P < 0.05). However, when frozen-thawed embryos were first cultured in vitro for 12 h, then exposed to 1.00 x, 0.50 x, 0.30 x, 0.25 x and 0.2 x PBS, the survival rates were 98%, 94%, 82%, 58% and 26% respectively. There was no statistical difference between 1.00 x and 0.50 x PBS group (P > 0.05). Although the rate in 0.30 x, 0.25 x and 0.20 x PBS group was significantly lower than in 1.00 x group(P < 0.01), it was significantly higher than in the same treatment group without in vitro culture(P < 0.05). PMID- 12549040 TI - [Mutation analysis of the tumor suppressor gene PTEN/MMAC1/TEP1 in human hepatocellular carcinoma]. AB - PTEN/MMAC1/TEP1 is a tumor suppressor gene. Its mutation has been found in several different types of human cancers. 34 primary human hepatocellular carcinomas have been examined for mutations in exon 5 and exon 8 of the PTEN gene. Exon 5 and exon 8 were amplified by polymerase chain reaction (PCR) with intronic primers and subjected to single strand conformation polymorphism (SSCP) analysis. SSCPs were found in 4 of the 34 hepatocellular carcinomas analyzed. Direct sequencing of the PCR products identified single base-pair substitutions in the four tumor DNA samples, two in intron 4 and two in exon 8. One of the base pair substitution in exon 8 is a missense mutation, which changed codon 304 of PTEN protein from Cys to Gly. These data suggest that PTEN may be involved in the carcinogenesis and development of hepatocellular carcinoma. PMID- 12549041 TI - [Cloning, expression of a segment of hTERT gene and detection of telomerase and hTERT by anti-hTERT polyclonal antibody]. AB - Telomerase is an important biomarker in cancer cells. It is active in germline cells, most of cancer tissues and cell lines, but not in most somatic tissues. Telomerase is composed of two components, and while hTER is present in normal and tumor cells, expression of hTERT appears to be highly regulated and correlates with telomerase activity. In order to detect the telomerase enzyme and hTERT protein, anti-hTERT polyclonal antibodies were produced in this study. A segment of hTERT cDNA was amplified by RT-PCR and cloned into the multi-cloning site of the GST gene fusion vector pGEX-5X-3. After the recombinant plasmid was expressed in E. coli BL21, the fusion protein was purified for immunization. Extracts from several cultured cells were analyzed by Western blot, and the results indicated that telomerase enzyme and hTERT protein could be specifically detected by this anti-hTERT antibod'. Thus, a simple and effective method was primarily established for the immunodetection of telomerase enzyme and hTERT protein. PMID- 12549042 TI - [Effects of GDNF on primary culture of spinal cord neurons]. AB - The effect of GDNF on long-term cultured spinal cord neurons was studied. GDNF could promote spinal cord neurons survival after 7 d or 14 d culture by MTT assay. The effect of GDNF on growth cones, neuron soma magnitude, neurite length and spines formulation of spinal cord neurons in cell culture was observed by phase microscopy, Nissl stain and NSE immunocytochemistry stain. The results indicated that GDNF had significant trophic effects on long-term cultured spinal cord neurons. PMID- 12549043 TI - [Salt stress-induced apoptosis in plants--a possible resistance mechanism to salt stress]. AB - Previous study has showed that apoptosis-like cell death can be induced under moderate salt stress in plants. In this paper, we studied the cell death induced by salt stress in maize, rice and tobacco roots utilizing DNA Laddering, paraffin sectioning-based TUNEL and chromosome spreading-based TUNEL simultaneously. The characteristic morphological and biochemical features showed apoptosis induced by salt stress may be a universal phenomenon in plants, but some differences may lie in various species. These results provided a valuable insight into studying the physiological mechanism of stress resistance in plants. In addition, we compared the in situ labelling technique based on chromosome spreading with that based on paraffin sectioning. We proposed according to the results that chromosome spreading-based in situ labeling technique should be suitable to detect individual cell death qualitatively and quantitatively with high efficiency, low background and detailed description of apoptotic changes at chromosome, nuclear and DNA levels. PMID- 12549044 TI - [The effect of integrin-like proteins on pollen germiantion and tube growth of Nicotiana tabacum]. AB - The effects of antiserum against human VnR integrin and integrin antagonist GRGDSP peptides on Nicotiana tabacum pollen germination and tube growth both in in vitro and in semi-vivo conditions were studied. No obvious inhibitory effects on pollen germination and tube growth in vitro were observed when anti-VnR serum or GRGDSP peptides was added to BK culture medium, but the enhancement of pollen germination and tube growth in vitro promoted by calmodulin was depressed by adding anti-VnR serum or GRGDSP peptides to BK culture medium. In addition to that, pollen germination and tube growth on stigma, as well as tube growth in styles were also inhibited at some extent by treating stigma and microinjecting GRGDSP peptides or anti-VnR serum into styles. The role of integrin-like proteins in regulation of pollen germination and tube growth in situ was discussed. PMID- 12549045 TI - [Inhibitory effects of N-(4-hydroxylphenol) retinamide on transformed human bronchial epithelial cells in vitro and reconstructed human bronchial epithelium in vivo]. AB - Precancerous lesion is one of most important steps in tumorigenesis. It has been shown that retinoids have reliable effects on controlling many kinds of animal tumor and malignant tumor cell lines in vitro, but there is no laboratory report on the biological effect of retinoids on the precancerous lesion of human lung cancer. In this study the methods including of cell serum-free culture, precancerous model of human bronchial epithelium reconstructed in rat trachea/xenotransplanted in nude mice, flowcytometry, immunohistochemistry, TUNEL and pathological observation have been used to study the biological effects of N (4-hydroxylphenol) retinamide (4-HPR), one new kind of retinoids, on transformed human bronchial epithelial cells in vitro and premalignant human bronchial epithelium in vivo. The results showed that in the study in vitro, the proliferation of transformed human bronchial epithelial cells, the ratio of cells in S phase, and the percentage of cells that positively react to antibody Ki-67 and mpm-2 were inhibited, but apoptotic cells were induced significantly by 4-HPR exposure. At the experiment in vivo, both growth rates and precancerous grades of the reconstructed human bronchial epithelium were reduced, and apoptotic cells were also observed in epithelium after 4-HPR treatment. The results suggested that 4-HPR is one of hopeful chemopreventive medicines to lung cancer. PMID- 12549046 TI - [Studies on the distribution of actin during meiotic prophase I of Nicotiana rustica L]. AB - The Pollen Mother Cells of Nicotiana Rustica L. were studied using the techniques of immunofluorescence microscopy and protein A-colloidal gold IEM. Immunofluorescence microscopy indicates the presence of actin in both nuclei and cytoplasm. IEM observation shows that gold particles are present in cytoplasm, chromatin and cytomictic channels. These results indicate that actin has some relation with chromatin condensing at synizesis. Moreover, actin may also play an important role in cytomixis. PMID- 12549047 TI - [The establishment of two new evaluation criteria for cell state of plant cell suspension cultures]. AB - An experiment on suspending rice cell cultures in sucrose solution and on optical density (OD) value in culture solution at 576 nm wave width was carried out, the result indicates that suspension rate (%) of suspensions, while suspending in 0.76 mol/L sucrose solution for 15 minutes at room temperature, can be employed as an judgment criterion of cell state. Suspending cell suspension in 0.76 mol/L sucrose solution is also a good approach on selecting cells from suspension cell mass, as a result, it is beneficial to establishing cell suspension. The browning of cell suspension is related to the necrosis degree of suspension cells. OD value of cell suspension liquid at 576 nm can be taken as a criterion with which the necrosis degree of cell suspension can be evaluated. PMID- 12549048 TI - [PCR-RFLP analysis of the genus Saccharomonospora]. AB - PCR-RFLP, a molecular method of polyphasic taxonomy of the genus Saccharomonospora was introduced. The feasibility of the application of this method at the species level in the taxonomy and identification of Saccharomonospora were discussed. The results showed that PCR-RFLP analysis is valuable for the determination of taxonomic position in the genus of some Saccharomonospora strains. PMID- 12549049 TI - [Expression and synergism of two cry insecticidal protein genes in Pseudomonas fluorescens]. AB - Several engineered Pseudomonas fluorescens(Pf) strains were constructed mainly based on a Pseudomonas plasmid pJMS6 alpha-lac and two insecticidal crystal protein genes of Bacillus thuringiensis, cry1Ac and cry2Aa, and the host Pf strain, P303, which was with highly antifungal activity to some plant disease fungi and colonizing ability on a wide range of plants. The DNA introduction was confirmed by PCR-RFLP and Southern blot. The 132 kD insecticidal protein was detected in IPP101 and IPP202 by SDS-PAGE and rhombic insecticidal protein crystals of them were observed through electron microscope, also indicating that cry1Ac gene was highly expressed. The results of insecticidal bioassay indicated that IPP101 was more toxic than IPP201, and IPP202 was the most toxic among the 3 strains. LC50 to the neonates of cotton boll worm(Helicoverpa armigera) were 0.02604, 0.00812 and 0.00186 mL/g feed, consecutively. In IPP202, two gene products showed significant synergism, with the co-toxicity coefficient of 332.8. PMID- 12549050 TI - [Cloning of a new catechol 1,2-dioxygenase gene (tfd C) from Plesiomonas and its expression in the E. coli]. AB - A new catechol-1,2-dioxygenase gene (tfd C) was cloned from the Plesiomonas using the PCR method. Primers were designed according to the reported sequence of Catechol-1,2-dioxygenase (C120) gene from Alcaligenes eutroplus. The amplified fragment contained a 765 bp open reading frame (ORF), encoding a protein of 255 amino acids. The new tfd C gene shared a high homology with the one cloned from Alcaligenes eutroplus, showing only one base difference at 693 site (C-->A) and consequently one amino acid difference at 228 site (P-->T). The ORF was cloned to the plasmid pBluescriptII KS, which was transferred to E. coli JM109 and a positive clone, pBt2G, was then selected. A significant activity of C120 was detected in the positive clone. When the ORF was cloned to the plasmid pET-30a, which was transferred to E. coli BL21(DE3) plysS, the expected 33 kD protein was detected from a positive clone, pET30A, by SDS-PAGE. The C120 is a key enzyme in degrading aromatic pollutants in the environment. In order to use plants to degrade aromatic pollutants, the gene will be introduced into the turfgrass. To express the gene properly in plants, its translation initiation codon was modified from GTG to ATG. A similar activity of C120 was obtained following the modification. PMID- 12549051 TI - [Cloning and expression of caiDE genes in E. coli BL21(DE3)]. AB - The caiDE genes for carnitine racemase and its relative factor were cloned into pUC18 from the chromosomal DNA of E. coli MC4100 by shot-gun approach and have been verified by sequence analysis. The target plasmid was named as pJX393 and subcloned to form a transcription plasmid pDSW2. After its transformation into BL21(DE3) strain and induction by IPTG, the recombinant strain containing pDSW2 expressed two apparent protein bands on SDS-PAGE, whose molecular weight were 30 kD and 24 kD individually. PMID- 12549052 TI - [Cloning and expression of Shiga-like toxin type II variant B gene of E. coli]. AB - A structure sequence and a DNA fragment including the signal peptide sequence and structure sequence of Shiga-like toxin II variant B subunit gene were amplified from E. coli strain O138 by PCR. After digested with restriction endonuclease EcoRI and BamHI, the two genes were orientally inserted into the polycloning site of expression vector pYA3334 (asd+) respectively. Recombinant plasmids pB0 and pB1 were constructed and amplified in E. coli X6212 (asd-). pB0 and pB1 were then introduced into avirulent Salmonella typhimurium vaccine strain X4550 (asd-) by serial transformation through intermediate strain X3730 (asd-) to construct recombinant SLT-IIvB strain. Results of nucleotide sequencing of the cloned fragments in pB0 and pB1 revealed that they were in correct ORF of SLT-IIvB. The results of SDS-PAGE and Western-blot showed that 7.6 kD protein of SLT-IIvB antigen was expressed at pretty high level in recombinant strain X4550(pB0). The results of mice immunization indicated X4550(pB0) could initiate the host to produce specific antibodies to SLT-IIvB and LPS-O antigen of X4550. So the recombinant strain X4550 (pB0) is worth considering as a candidate vaccine strain against porcine edema disease and Salmonella typhimurium infections. PMID- 12549053 TI - [Studies on sanH and sanI--genes related to nikkomycin biosynthesis of Streptomyces ansochromogenes]. AB - A 7.0 kb DNA fragment was cloned by reverse genetics, which was located in the gene cluster of nikkomycin biosynthesis of Streptomyces ansochromogenes. Besides sanF--a gene related to nikkomycin biosynthesis of S. ansochromogenes, the sequence of a 2.2 kb BglII DNA fragment upstream sanF was determined. This 2.2 kb DNA fragment cointains two complete open reading frames (ORF). ORF1 consists of 1233 base pairs, ORF2 consistis of 195 base pairs, and they encode proteins with 410 amino acids and 64 amino acids respectively, which were designated as sanH and sanI. In search of databases, the deduced products of sanH has 46% amino acid identities and 62% amino acid similarities in comparison with the cytochrome P450 of Streptomyces griseolus, and the deduced protein of sanI has 56% amino acid identities and 70% amino acid similarities with the ferredoxin of S. griseolus. The function of sanH gene was studied using strategy of gene disruption, and the result showed that the disruption of sanH didn't influence nikkomycin biological activity in S. ansochromogenes. PMID- 12549054 TI - [The construction and application of Streptomyces-E. coli shuttle plasmid pSGLgpp]. AB - The high-copy-number plasmid pSGL1(7.4 kb) was isolated from Streptomyces geobisporous. Its minimal replicon had been determined and sequenced. With the total DNA of S. geobisporus as template, the DNA fragment involving C1027 apoprotein signal peptide encoding sequence (gpp) was amplified by PCR. After this fragment was inserted into the plasmid pSGLN, a derivative plasmid of pSGL1, one new Streptomyces-E. coli shuttle plasmid was obtained namely as pSGLgpp. Using the new vector, we carried out the expression of hsIL-1RI in streptomyces. PMID- 12549055 TI - [Construction of cDNA library of Mortierella and screening of delta 9 fatty acid desaturase cDNA sequence]. AB - Mortierella species have potential for fermentative production of polyunsaturated fatty acids including gamma-linolenic acid, Arachidonic acid and EPA, etc. In order to clone genes encoding enzymes in the unsaturated fatty acid biosynthetic pathway, cDNA library of Mortierella was constructed using lambda gt 10 vector. Using cDNA encoding conserved region of delta 9 fatty acid desaturase gene as probe, Mortierella cDNA library was screened. After two rounds of screening one positive clone was identified which has insert length of larger than 1.6 kb. PMID- 12549056 TI - [Nucleotide sequence analysis of E2 major protective antigen encoding region of 12 strains of hog cholera virus(HCV)]. AB - cDNA fragments, of HCV envelope glycoprotein E2 major gene of 11 field strains isolated in China in different time and 1 French reference strain(Thiveral) were amplified respectively with RT-PCR method and sequenced. The fragments amplified located by the 5' 2485 to 2708 of E2 major domains B and C and encoded 75 amino acid residues of E2 glycoprotein. All the products amplifield by RT-PCR from 12 strains in the study were same size of 224 bp. Comparing 12 sequences with other 9 references strains sequence reported before using software DNAstar, it was found that Hog Cholera Virus could be classified in two groups by analysis of phylogenetic tree. Strains Brescia, Gpe, Ald, Thiverval, C, CW, HCLV, HCVSM, BJCY1/96, BJTX3/96, BJSY2/96, HeNXH2/98, HeNZZ1/82 and GDGZ1/95 were assigned to group A and were 85.7%-100% for nucleotide sequence and 83.8%-100% for amino acid sequence in homology; but strains HCVF98, HCVF94, HeBHH2/95, LN1/84, SZGM1/85, SCCD1/79 and Alfort were assigned to group B and were 84.3%-100% for nucleotide sequence and 85.1%-100% for amino acid sequence in homology; and 21 strains of HCV were 78.1%-100% for nucleotide sequence and for 78.4%-100% amino acid sequence in homology. Homology were 99.1% for nucleotide sequence and 100% for amino acid sequence between strains HCLV in our study and strains C reported by Rijn's in the Netherlands. It's showed our method of sequencing is reliable. There were obvious differences between the two groups in sequences of envelope glycoprotein E2 major gene, especially in the amino acid substitutions of sites 713 and 729 respectively, and it is showed the two groups of HCV field strains might vary genetically in some extents. The results of other report of challenging of the partial field strains showed that the Chinese stock vaccine virus(HCLV) has good immunity. PMID- 12549057 TI - [Studies on the subgenomic RNAs of hepatitis E virus]. AB - Through labeling HEV RNAs with [32P]-phosphate, two subgenomic RNAs at 6.5 to 7.5 h post-infection, one subgenomic RNA at 8.5 to 10.5 h post-infection and none subgenomic RNAs were detected at 21 post-infection or in purified virus. Hybridization results illustrated that the HEV subgenomic RNAs form a 3' nested set which is situated at the 3' end of genome, but their 5' ends do not share a common leader sequence. UV transcription results showed that the HEV subgenomic RNAs were transcribed independently from its promote. Primer extension analysis suggested that the 5' end of large subgenomic RNA was mapped to the initiative region within RNA polymerase region, and the small subgenomic RNA was mapped to the intergenic region between ORF1 and ORF2. PMID- 12549058 TI - [Producing characterization of constitutive-form laccase by Basidiomycete]. AB - A constitutive-laccase-producing fungus was gained from 20 strains of different sources with plate method. Laccase activity reached 0.58 IU/mL when the strain grew on DC medium with shaking cultivation, which was superior to stationary method by 4 times. The enzyme production paralleled with biomass in early 8 days, subsequently activities declined slightly though its biomass continued upward. Tyrosine and tryptone at the concentration of 4 g/L improved the enzyme production, reaching 2.49 and 2.0 IU/mL respectively. Some tested chemical agents seemed not to contribute to the increase of the enzyme production effectively except for syringaldazine and o-tolidine. Heatshock at 40 degrees C for 1 h made 1.3 times more enzyme expression. PMID- 12549059 TI - [ATP synthesis by free yeast catalysis coupling hollow fiber ultrafiltration separation]. AB - A novel process for the ATP production employing free yeast catalysis and coupling a hollow fiber module ultrafiltration separation was studied. The experimental results of biocatalyst half-life and operating stability for ultrafiltration showed that the enzymes released from free yeast cell could be cut-off efficiently from the product output stream by hollow fiber membrane. The intercepted percentage of ADH and HK reached more than 95% during the steady running. The efficiency of enzymes utilized was 2.0-2.5 fold as much as that of batch reaction. And the time of yeast cells used repeatedly for ATP synthesis reached 2.5-3.0 batches. Under 0.1 MPa, the ultrafiltrating speed of hollow fiber membrane could keep on a stabilizing value for 11 batches continuously. The continuous operation mode of ATP synthesis simultaneous product ultrafiltration separation could be maintained in higher conversion rate for 5.0 h when the whole system fell in a pseudo-stat where the dilution rate was equal to 0.25 h-1. PMID- 12549060 TI - [Studies on monoclonal antibodies of Magnaporthe grisea and its interfering effect on appressorium formation]. AB - Selecting by indirect ELISA eleven hybridoma cell lines, secreting monoclonal antibodies (McAbs) against Magnaporthe grisea were produced by fusing mouse myeloma cells(SP2/0) with spleen cells from BALB/c mice immunized with the mixture of conidia, germ tubes and appressoria of M. grisea using 50% PEG. The result of IFTC test showed that four of McAbs, named as 2B4, 4A1, 1D1, and 2H4 specifically bound to the cell wall surface of the fungus; Western blotting revealed that 2B4, 4A1, 1D1 were recognized different protein antigens from the surface of conidia and germ tubes; These four McAbs could effectively interfere with the appressorium formation both on cellophane membrane and the surface of onion epidermis and inhibit the disease leaf lesions development in vitro test. PMID- 12549061 TI - [A actinomycete strain C3905 producing antitumor antibiotic sandramycin from Antarctica]. AB - A nocardioform actinomycete strain C3905 was isolated from a soil sample collected in King Geoge island, Antarctica. Its aerial mycelium is white, substrate mycelium is colorless to cream or pink. The hyphae of mycelium 0.5 micron-0.8 micron in diameter and they fragment into irregular rod to coccoid elements with smooth surface. Cell wall type I. The mycolic acids are lacking. Mk 9(H4) is the predominant menaquinone component. Phospholipid type PI. The G + C content of DNA is 68.3-68.9 mol%. Facultative psychrophile. The optimum temperature for growth is 15 degrees C-20 degrees C. Produces antitumor antibiotic sandramycin. Based on the characteristics above as well as the investigation of the molecular genetics. We propose that strain C3905 should be placed in the Nocardioides albus as a variant, named Nocardioides albus var. antarcticus. PMID- 12549062 TI - [Isolation and identification of myxobacteria]. AB - This paper reported that the authors isolated large numbers of myxobacteria from more than 100 samples collected in many places of China. More than 400 pure strains were obtained. These strains belonged to 10 genera of Myxococcales. The most frequent genera isolated were Myxococcus, Sorangium, Corallococcus, and Cystobacter. Melittangium was seldom isolated. No Chondromyces and Haploangium were found. PMID- 12549063 TI - [The evolution of pathogen-host interactions mediated by bacteriophages]. PMID- 12549064 TI - [Cold-active enzyme produced by cold adapted microorganisms]. PMID- 12549065 TI - [How to take a better role for National Natural Science Funds in its support to applied basic research on microbiology]. PMID- 12549066 TI - [Relationship between endogenous active oxygen species, ethylene and the development and operation of the alternative pathway in tobacco callus under salt stress]. AB - This paper reported firstly the dynamic changes of the alternative pathway and the cytochrome pathway in tobacco callus under different concentration of salt. The production rate of endogenous ethylene, the contents of active oxygen species (O2-., .OH and H2O2) and the activities of related enzyme (SOD and CAT) were also investigated simultaneously. The results indicated that the actual operation of the alternative pathway (rho Valt) increased gradually and reached the maxima at 0.75% salt concentration, then declined, while the capacity of the alternative pathway (Valt) didn't exhibit the same change pattern with rho Valt and the cytochrome pathway (rho' Vcyt) changed contrary to rho Valt. At the same time, H2O2 and O2-. accumulated rapidly and reached their peak value at about 0.5% and 0.75% salt concentration respectively, while the content of .OH increased consistently. The production rate of endogenous ethylene increased rapidly and reached its maxima at about 0.5% salt concentration. These data demonstrated that changes of Valt in tobacco callus were associated with a parallel production of H2O2 and endogenous ethylene and a opposite production of .OH, while rho Valt changed parally with the production of O2-.. On the basis of above results, it was deduced that the changes of the development and operation of the alternative pathway in tobacco callus under salt stress might be correlated with the production of endogenous ethylene and the active oxygen species. The possible participation of the alternative pathway in response to salt stress was also discussed. PMID- 12549067 TI - [Expression and characterization of t-PA in insect cells]. AB - With the RT-PCR technique, human tissue-type plasminogen activator (t-PA) cDNA was amplified from human melanoma cells. The sequence was proved to be the same as those reported by foreign researchers. The t-PA cDNA containing whole reading frame was inserted into the transfer vector pBacPAK8. The constructed pBac-tPA and the linear BacPAK6 virus DNA were cotransfected with Tn-5B-1 insect cells by lipofectin-mediated transfection method. Eleven pure recombinant viruses were isolated by plaque assay, and one of them, nominated BactPA3, was selected by PCR identification and biological activity comparison. The t-PA activity expressed in serum-containing media reach the highest level at 72 h postinfection. The activity was 3.04 x 10(3) IU/ml, e.g. 1.8 x 10(4) IU/10(6) cells. The highest expression level in serum-free media was almost the same, but needed more time (at 132 h postinfection). SDS-PAGE fibrin autography showed that the molecular weight of the expressed t-PA was about 68 kDa. Its stimulation by fibrinogen, affinity with fibrin and inactivation in plasma were almost the same as the native t-PA purified from human melanoma cell culture. The half-life of t-PA expressed in serum-free media was seven minutes. PMID- 12549068 TI - [Prokaryote expression and western analysis of BcpLH gene of Chinese cabbage]. AB - BcpLH gene preferentially expressed in folding leaf of Chinese cabbage contains dsRNA-binding domains. The cDNA of BcpLH gene was cloned into a His-fusion expression vector pET-28a (+) and was induced to express in E. coli strain BL21 (DE3). Then, the specific protein was partially purified and the rabbit was immunized to prepare the anti-serum. Meanwhile BcpLH cDNA was cloned into the pMAL-c2 containing the solubizing partner, and then the soluble protein generated. It was demonstrated from Western dot assay that the BcpLH protein was specific. The BcpLH active protein and its anti-serum made it possible to study RNA-binding activity and regulation mechanism in plant development. PMID- 12549069 TI - [Antibacterial and antifungal effects of Agkistrodon halys Pallas: purification of its antibacterial protein--LAO]. AB - This paper reports the venom from Agkistrodon halys Pallas have inhibitory activity against fungi and E. coli by tested in a disc diffusion assay. An antibacterial component--LAO from the venom were purified to homogeneous. It had not only antibacterial effect, but L-amino acid oxidase activity. And its enzymatic specific activity was 808 U/mg. The venom had at least 3 antibacterial components (I, II, III) as determined by acid polyacrilamide gel electrophoresis, LAO is the antibaterial components II. PMID- 12549070 TI - [Study of chromosome pellicle by indirect immunoflourescence method and antigen analysis in Vicia faba]. AB - The autoimmune antiserum specific to pellicle of human metaphasic chromosomes from a lupus patient was used to stain metaphasic chromosome Vicia faba by means of indirect immunoflourescence method. It was found that the pellicles of vicia metaphasic chromosomes was positively stained. The antigen of Vicia faba recognized by the antiserum was also examined by PAGE of total cell lysate and western blotting. PMID- 12549071 TI - [Production of intergeneric somatic hybrid plants via protoplast electrofusion in citrus]. AB - Leaf derived protoplasts of Sour orange (Citrus aurantium L.) were fused electrically with embryogenic protoplasts of Microcitrus papuana Swingle. Plants were regenerated from the fusion products, which were characteristic with three types of leaf morphology. Most of the plants were identical to Sour orange (namely, Leaf-parent-type plant) and two plants had large and thick leaves whereas one plant had bifoliate and trifoliate leaves. Chromosome examination showed that these plants were diploid with 18 chromosomes (2n = 2x = 18). RAPD analysis was employed to verify the hybrid characteristics of the plants in the first two types. Four 10-mer arbitrary primers with polymorphism were chosen. Band pattern of the plants was similar with the leaf parent (Sour orange) for the primer OPAA-17. Band pattern of the plants was similar with either Sour orange or M. papuana for OPA-08. As for OPA-07 and OPA-04 three kinds of band profiles were detected. Results of RAPD marker, together with chromosome determination, indicated that all of the analyzed plants were intergeneric diploid somatic hybrids between Sour orange and M. papuana. PMID- 12549072 TI - [TGF-beta 1 rapidly reduced hepatocarcinoma cells adhesion on to fibronectin and stimulated FAK dephosphorylation]. AB - In order to investigate whether TGF-beta 1 could rapidly regulate integrin induced signaling, we treated SMMC-7721 human hepatocellular carcinoma cells with human recombinant TGF-beta 1 for 10 min, and examined cell adhesion, integrin amount and FAK tyrosine phosphorylation. We used cell adhesion assay to estimate the affinity of alpha 5 beta 1 integrin with fibronectin, and analyzed the amount of integrin alpha 5 and beta 1 subunits by performing FACS analysis. Then western blot analysis was carried out to examine tyrosine phosphorylation level of FAK. Our results showed that TGF-beta 1 could rapidly attenuated cell adhesion onto Fn without changing the expression of alpha 5 beta 1 integrin, and at the meantime dephosphorylated FAK. It suggested that TGF-beta 1 rapidly regulated the activation of integrin, and stimulated FAK dephosphorylation, which might induce depolarization in SMMC-7721 hepatocellular carcinoma cells, then facilitates the detachment of tumor cells at early stages of migration. PMID- 12549073 TI - [Study on expression and resistance of the double drug resistance genes transduced into human umbilical cord blood CD34+ cells mediated by bicistronic retroviral vector]. AB - To explore whether human umbilical cord blood CD34+ cells transduced with human aldehyde dehydrogenase class-1 (ALDH1) and multidrug resistance gene (MDR1) increase resistance to 4-Hyaroxycyclophosphophamide (4-HC) and P-Glycoprotein Effluxed Drugs, a bicistronic Retroviral vector G1Na-ALDH1-IRES-MDR1 was constructed. The vector was transduced into the packaging cell lines GP + E86 and PA317 by LipofectAMINE. Using the medium containing VCR and 4-HC for cloning selection and pingponging supernatant infection between ecotropic producer clone and amphotropic producer clone, we obtained high titer amphotropic PA317 producer clone with the highest titer up to 5.6 x 10(5) CFU/ml. Cord blood CD34+ cells were transfectced repeatedly with supernatant of retrovirus containing human ALDH1 and MDR1cDNA under stimulation of hemopoietie growth factors. PCR, RT-PCR, Southern blot, Northern blot, FACS and MTT method analyses show that dual drug resistance genes have been integrated into the genomic DNA of cord blood CD34+ cells and expressed efficiently. The transgenes recipient cells confered 4- to 7.2-folds stronger resistance to cyclophospsphamede and P-Glycoprotein Effluxes drug in comparison with the nontransduced cells. This study provided a foundation for the application of combination chemotherapy in tumor clinical trial. PMID- 12549074 TI - [Ultrastructure differences of in vitro cotton fiber and native cotton fiber]. AB - Native cotton fiber and in vitro cotton fiber that was induced from cotton ovule callus by suspension culture were observed using transmission electron microscope and scanning electron microscope. The ovule surface on the first day preanthesis was quite smooth. On the anthesis, it had a lot of protuberances. Two kinds of callus, smooth and rough were found. The microfibrils of callus was vertical to the cell long axis and they changed their orientations with the development of the in vitro cotton fiber: from the vertical to shallow spiral and then to parallel to the cell long axis. So was the native cotton fiber. It suggests that in vitro cotton fiber and native cotton fiber have similar development process. Compared with the ovule surface cell, most callus cells had smaller nuclear. During the development of the fiber, the plasm of native cotton fiber was denser than that of in vitro fiber, and it has more cellular organ than in vitro fiber. The cell wall of native cotton fiber was thicker and denser than that of the in vitro cotton fiber too. It suggests that the physiological activity of in vitro cotton fiber was less active than native cotton fiber. PMID- 12549075 TI - [The somatic embryogenesis and establishment of transformation experiment system in Larix principis-Rupprechtii]. AB - Larix principis-Rupprechtii is one of the superior afforestation forest trees growing in north China. Embryogenic cultures were initiated from immature zygotic embryos of Larix principis-Rupprechtii on S culture medium containing 2, 4-D 0 2.2 mg/L, KT and BA each at 0-0. 8 mg/L. Embryogenic calli were subcultured and multiplicated on S + B culture medium containing dropping off each hormone concentration. We set up 33 steady-going embryogenic cell lines; We studied on the growth stage and genotype differences of every embryogenic cell lines; and Finded more than 10 high-frequency somatic embryogenesis cell lines such as 2K, 2T, 2I, 2J, 3C etc.. The number of 2T somatic embryos reaches 314/per gram of embryogenic tissue and the number of 3C somatic embryos is 185/per gram of embryogenic tissue. The re-induction method of Larix principis-Rupprechtii from somatic embryos was used to produce renewable embryogenic cultures and steady going embryogenic cell lines effectively. Mature somatic embryos can germinate and develop further into plantlets when they are isolated and cultured on a hormone-free WPM culture medium. The regeneration plantlets were obtained. Furthermore, the transformation with a truncated gene of Bacillus thuringensis (B. t) were carried out, the PCR showed positive results, because of this, embryogenic cell line of Larix principis-Rupprechtii can be used for transformation experiments to support further breeding in forestry. PMID- 12549076 TI - [Studies on the Robertsonian translocation of Microtus mandarinus]. AB - The type of chromosome No. 1 and chromosome number from 53 individuals of Microtus mandarinus have been studied and compared in three sex types: XY, XX, XO. We found that the first pair of autosomes are very unstable, and there are three types: (1) M, M (With a double metacentric chromosome), (2) M, T, T, (With single metacentric chromosome). (3) T, T, T, T (Without metacentric chromosome). The chromosome number of the same sex individuals changes regularly with the type change of chromosome No. 1, that is, the increase of one chromosome in 2n number is always accompanied by the increase of two T and the decrease of one M, and vice versa. The synaptonemal complexes (SCs) of spermatocyte in pachytene nuclei from the males (2n = 51) were analysed by the electron microscopy. The SCs studies demonstrate that there are 23 fully paired autosomal bivalents, XY bivalent and an autosomal trivalent. This trivalent is formed by one metacentric and two telocentric elements and characterized by the presence of two short side arms. Meanwhile, all trivalents are in a cis configuration. The study of G banding also demonstrates that the No. 1 autosome polymorphism is caused by Robertsonian translocation. Robertsonian fission is the main reason of the polymorphism of chromosome No. 1 and of variation of chromosome number in M. mandarinus. PMID- 12549077 TI - [Study on reclassification of extremely thermoacidophilic archaea strain S5]. AB - The further study on thermoacidophilic archaea strain S5, with has been identified as Sulfosphaerellus thermoacidophilum gen.nov.,sp.nov, has shown it was able to grow facultatively aerobically by means of two sulfur-metabolizing modes of chemolithotrophy which is the characteristic of Acidianus. And the 16SrRNA gene of strain S5 was amplified, cloned and sequenced, a phylogenetic tree was constructed on the 16SrRNA gene sequences. The tree clearly indicated that strain S5 formed the same lineage with Acidianus brierleyi. Thus strain S5 should be the member of Acidianus. However, there are only 44%, 22% and 23% genomic DNA similarity between S5 and A. brierleyi. A. infernus and A. ambivalens, respectively. And the G + C content of S5 DNA is 38%, which is 5% ~ 7% higher than the reported G + C contents of the other Acidianus species (31% or 32.7%) . In addition, strain S5 is a strictly chemolithoautotrophs, which is obviously different from facultative chemolithotrophs of A brierleyi. Based on the observed differences, strain S5 represents a new species within the genus Acidianus. A new species name, Acidianus tengchongenses, was proposed for it. The type strain is designated S5. PMID- 12549078 TI - [Characterization and phylogenetics of a new species of genus Methanobacterium]. AB - A strain of methanogenic bacterium Px1 was isolated from an anaerobic digester in the department of environmental sciences, TsingHua University, which treated the wastewater from Beijing Beer Factory. The isolate is curved rod, light-yellow colony, uses only H2 + CO2 as the substrate for CH4 formation. On the basis of physiology, morphology, structural characteristics and phylogenetic analysis, it is turned out that strain Px1 is a new species of the genus Methanobacterium, the name Methanobacterium curvum sp. nov. is proposed. PMID- 12549079 TI - [Study on the application of quantitative analysis of cell-wall amino acids in actinomycetes classification]. AB - This paper has collected 70 recognized type strains of actinomycete from home and abroad Microbiological culture collection centers. Adopting thin-layer chromatography and thin-layer chromatogram scanner methods to quantitatively analyze the composition of cell-wall amino acids in actinomycete. It has made comparison between quantitative results and qualitative ones. After a thorough discussion, a revision is made on the standard of dividing cell-wall types in actinomycete chemical classification, which has provided some original suggestion in hope of improving the research of actinomycete chemical classification. PMID- 12549081 TI - [Introduction of the chromogenic gene to the plant growth-promoting rhizobacteria of cucumber]. AB - Using a bicomponent transposition system with the E. coli lacZY gene cloned between Tn7 termini, a sensitive, selectable marker based on expression of the E. coli lac operon genes encoding beta-galactosidase and lactose permease was transformed into the rifampicin resistant mutant of plant growth-promoting rhizobacteria of cucumber, Pseudomonas aeruginosa CN116 and Pseudomonas corrugata CN31, respectively. Transformants were conferred the ability to utilize lactose as a sole carbon source and the ability to cleave the chromogenis substrate X-Gal to show a specific blue color. Southern blotting analysis showed that lacZY gene was inserted into the genome DNA of target strains. Compared with the wild type strains, the cultural characters, morphological features, growth promoting and disease control effects of transformants were almost unchanged, except the new marked phenotype. This marker system enabled the detection of lac+ transformants at sensitivity of 10 CFU/g soil, which makes the further studies on PGPR more easily. PMID- 12549080 TI - [Studies on the isolation, culture and DNA identification of mycelia of Tricholoma matsutake]. AB - The tissue isolation for Tricholom matsutake(S Ito et Imai) Sing were made with 8 media in 810 test tubes from different positions of 9 basidiocarps of different source and from mycorrhizae, and soil with the fungi in the studies. The results showed that 94 test tubes of slow-growing mycelia were isolated from lamellae and their success percentages of isolation with media PDAS, PDAW, BM, PDA were 74.4%, 355%, 15.6% and 8.9% respectively. The fast-growing mycelia were easily got from the mycorrhizae and soil related to matsutake. The isolates with different culture characteristics were appraised through DNA fingerprinting comparison with matsutake basidiocarps collected from Jilin province, China and reference isolates presented by matsutake research workers of China and Japan, in which RAPD (Random Amplified Polymorphic DNA)-PCR patterns were sharply prepared using 17 arbitrary decamer nucleotide primers screened. The statistical data indicated that all slow-growing mycelia isolated from lamellae had the same DNA fingerprinting patterns as their origin basidiocarps tissues such as pileus (containing lamellae) and stipe, whose similarity coeffecients all were 1.000, and were therefore identified as true Tricholoma matsutake. However, the fast growing mycelia or yeast colony were identified as not matsutake. The results suggested that matsutake and its own mycelia have DNA homogeneity, and there exists no any other microbe in the basidiocarps. The results also demonstrated that all matsutake from east China and reference isolates of matsutake from southwest China and Japan were one same species Tricholoma matsutake, whose DNA similarity coeffecients varied from 0.934 to 0.994. PMID- 12549082 TI - [Identification and location of the toxin protein genes in 56 Bacillus thuringiensis isolates]. AB - Polymerase chain reaction was performed to screen Bacillus thuringiensis isolates for cryI, cryIII, and cryV genes with cryI-, cryIII-, and cryV-specific primers respectively. The results indicated that, in the 56 Bt isolates examined, 7 isolates contain cryI gene, 2 isolates contain cryIII gene, and 21 isolates contain both cryI and cryV genes. The cryV gene was located by Southern hybridization in the large plasmid of about 150 MD molecular mass. PMID- 12549083 TI - [The detection of phenol degrading strain in environment with specific primer of phenol hydroxylase gene]. AB - A 684 bp oligonucleotide fragment was produced by PCR amplification from phenol degrading strain Acinetobacter calcoaceticus PHEA-2 with the specific primers of gene encoding phenol hydroxylase. The nucleotide sequence of this fragment and its deduced amino acid sequence share 84% and 98% homology with the phenol hydroxylase gene and its deduced amino acid sequences of phenol-degrading strain Acinetobacter calcoaceticus NCIB8250. Sets of different aromatic compounds degrading strains were used to test this specific prime. The 684 bp-fragments were amplified only from phenol-degrading strains by PCR. When using this pair of primers to detect the bacterial isolates from wastewater discharged from coking plant, all the tested strains, which possess 684 bp characteristic fragment, showed the ability to degrade phenol in this study. PMID- 12549084 TI - [Transcriptional activator PlcR regulate the expression of multiple genes in Bacillus cereus]. AB - Food-poisoning and some diseases of human can be caused by potential pathogenic factors including phospholipases of c-type, hemolysins, enterotoxins, emetic toxin and others from Bacillus cereus which is an opportunistic pathegon. Generally, the expression of pathogenic genes is completed by a co-regulation in many bacteria. The isolation of a mutant, displaying significant reducing expression of lecithinase and general proteolytic activities, from a transposon induced bank of B. cereus type strain ATCC14579T were described. Sequencing data revealed that a gene with high homology to transcriptional activator (plcR) of B. thuringiensis was inactivated by insertion in this mutant. The results suggest that plcR regulates the expression of multiple genes in B. cereus. PMID- 12549085 TI - [Cloning and expression of VHB gene in D-arabitol producing yeast]. AB - Recombinant plasmid pVgb-EX2 containing Vitreoscilla hemoglobin gene vgb and formaldehyde resistant gene SFA1 was constructed and transformed into D-arabitol producing yeast strain Saccharomyces sp. X-62. The fact that the amount of VHb in transformant cells was considerably higher than that in control cells indicated that gene vgb was expressed in transformant cells. D-arabitol productivity and yield of fermentation by transformants were improved. The most improvement of D arabitol productivity in repeat experiments reached 27.3%. It appeared that the fermentation productivity of D-arabitol was relative to the amount of VHb in cells under experimental conditions. PMID- 12549086 TI - [Sequencing of E2 gene and comparison analysis of four strains hog cholera virus (HCV)]. AB - Four cDNA fragments of envelope glycoprotoin E2 gene of SM strain, HCLV strain, F03 strain and F07 strain were amplified respectively with RT-PCR method. The amplified E2 fragments of four HCV strains were all 1273 bp in length by agarose gel electrophoresis. Four E2 fragments were cloned respectively into pGEM-T easy vector. 1273 bp cDNA fragment of Four HCV E2 gene were sequenced and 381 residues amino acid sequence of E2 glycoprotein were deduced. The signal peptide sequence (WLLLVTGA) located in the N-terminal residues 683-690 and the transmenbrane region (TMR) sequence located in the C-terminal residues 1031-1063 of Four E2 gene were highly conserved and hydrophobic region. The conserved sequence among the E2 protein of pestiviruses, RYLASLH which located in the N-terminal residues 753-759 of domains B and C, was hydrophilic, and none of them were variable among the pestiviruses, and the greatest values of antigenic in all E2 antigenic domain. This suggest that the conserved sequence RYLASLH are involved in E2 epitopes. The number and locations of 15 cysteine residues in E2 are conserved among pestiviruses, suggesting that the structure of this glycoprotein is similar in pestiviruses and the first six cysteines are critical for the correct folding of E2 and essential for all identified epitopes. It is showed epitopes. between HCLV strains and field strains are not variable obviously by analysising the varintion of some main amino acid residuse substitutions of E2 major antigenic domains. PMID- 12549087 TI - [Cloning and expression of the envelope glycoprotein gD gene of pseudorabies virus EA strain]. AB - The envelope glycoprotein gD gene of pseudorabies virus Ea strain was cloned via PCR technique. Sequence analysis displayed 98% nucleotide sequence homology and 97% deduced amino acid sequence homology between our cloned gD gene and PRV Rice strain gD gene. The recombinant transfer plasmid pSX35A-gD was obtained by inserting D gene into the baculovirus transfer vector pSX35A with whole-phase promoter cassette, then transfected insect cell Hi5 with linearized AcMNPV-OCC- virus DNA, and formed recombinant baculoviruses AcMNPV-OCC(+)-gD by homologous recombination in insect cell. Recombinant baculoviruses infected insect cell Hi5 after being purified by plaque assay. Both SDS-PAGE and Western-blotting showed glycoprotein gD with a molecular weight of about 47 kD was expressed specifically, product was about 6.2% of total cellular protein, and expressed gD was of immunogenicity. PMID- 12549088 TI - [Studies of the resistance of the recombinant alphavirus RNAs containing dengue-2 PrM gene to virus infection]. AB - The amplified PrM gene of dengue-2 virus was cloned into the downstream SP6 promoter-pSFV vector and the recombinant plasmid (pSF.rM2) DNA which contained sense- or antisense-PrM gene, was selected. pSF.rM2 DNA and helper DNA linearized by the enzyme SpeI digestion were both transcribed in vitro into recombinant RNAs which contained the capping analog on the 5'-end and contaansfected into BHK cells by electroportation. The the transfected host cells were challenged with dengue-2 virus and the resistant efficiency of recombinant virus RNAs containing sense- or antisense-PrM gene to virus infection were observed, respectively. The recombinant plasmids (pSFV-PrM) containing sense- or antisense-PrM gene were selected with determination of the nucleotide sequence. The recombinant virus particles were obtained with recombinant RNA and helper RNA co-transfected into BHK cells. Host cells transfected with antisense-PrM RNA derived complete resistance to dengue-2 virus replication and the efficiency was higher than that of the recombinant virus RNA containing sense-PrM gene. PMID- 12549089 TI - [Purification and characterization of a DNA helicase from the hyperthermophilic archaeon Sulfolobus shibatae]. AB - A DNA helicase from the hyperthermophilic archaeon Sulfolobus shibatae has been purified to homogeneity by column chromatography on Q Seqharose, phosphocellulose P11, heparin agarose, Superdex 200 and phenyl Superose. The purified enzyme shows DNA-stimulated ATPase activity. The molecular mass of the enzyme is approximately 63 kD. The S. shibatae DNA helcase is capable of unwinding a 70 bp duplex DNA flanked by unpaired single-stranded tails at both ends. The helicase activity requires Mg2+ and hydrolysis of ATP, and is inhibited by NaCl at concentrations greater than 200 mmol/L. It has an optimal pH of 6.7. The enzyme is active at temperatures between 40 and 80 degrees C, and the activity peaks at 70 degrees C. The S. shibatae DNA helicase is the first native DNA helicase to be isolated from archaea. PMID- 12549090 TI - [Composition and structure of bioflocculant BP25]. AB - A strain of Bacillus megaterium screened from activated sludge could produce exocellular flocculant. The flocculant was purified through ethanol precipitation and Sephadex S-500 column chromatography. The purified flocculant BP25 was assayed by Bradford reaction, agarose gel electrophoresis and Sulfate-phenol method. The results showed that BP25 was a kind of polysaccharide which contains 36.97% O, 6.28% H, 47.00% C, 0% No NMR assay showed BP-25 contains no uronic acid. Gas-chromatography assay combined with thin-layer chromatography of acid hydralate revealed that BP25 contains Glucose and Mannose with the mole ratio of 4:1. Methylation analysis revealed the polysaccharide contains alpha-1, 6 glycosidicbond and alpha-1, 3 glycosidicbond. The main chains are comprised of Glucose and all the Mannose are in the side chains. Possible repeating unit structure was deduced. PMID- 12549091 TI - [The epoxidation of cis-propenylphophonic acid to fosfomycin by Pencillium sp]. AB - One strain of Penicillium sp. F5, found from soil sample, was able to stereo specific epoxidation of cis-propenylphosphonic acid(PPOH) to fosfomycin[(-)-(1R, 2S)-1, 2-epoxypropylphosphonic acid, FOM] during the cultivation. The product (FOM) was identified by thin layer chromatography and microbiological assay. Under the culture conditions of 28 degrees C pH7.5 and 280 r/min, 0.3% concentration of PPOH, reached a level of 2.2 mg/ml of FoM and the yield was 41%. When the PPOH concentration was 0.6%, the yield was 8%. PMID- 12549092 TI - [Synthesis of galacto-oligosaccharides by immobilized Bacillus stearothermophilus]. AB - Bacillus stearothermophilus was embedded in sodium alginate, chitosan, gelatin respectively and utilized for galacto-oligosaccharides (GOS) production. By comparing with each other in the enzyme recovery, optimal reaction conditions, yields of GOS and mechanical strength of the carriers, the gelatin was selected as better carrier for immobilization. pH, temperature, lactose concentration, lactose conversion and mass transfer resistance of carrier significantly influenced the Yield of GOS. The maximum yield of GOS was 31.2% in a stirred reactor with lactose concentration at 60%. It remained about 88% after 96 h (8 batches) reaction. A packed bed was employed for a continuous reaction at space velocity 0.09 h-1, where GOS yield and reactor productivity was 31.5% and 17.4% respectively. It lost about 20% of its original yield of GOS after 140 h reaction. Products were separated by active charcoal columm. The chemical structure of tetrasaccharide identified by 13C-NMR was beta-D-Gal-(1-->3)-D-Gal (1-->6)-Dal-(1-->4)-D-Glu. PMID- 12549093 TI - [A study on DPPH free-radical scavengers from Xylaria nigripes]. AB - The DPPH free-radical scavengers from the fungal Chinese medicine 'Wulingshen' (Xylaria nigripes) were studied. After separation using silica gel colunm chromatography, MPLC and HPLC, 20 DPPH free-radical scavengers with purity higher than 85% and yield more than 2 mg were screened, in which compound B4-16 had relatively higher yield and stronger free radical scavenging activity. The formula of compound B4-16 was determined to be C10H10O4 (MW: 194) based on its FAB-MS, 1H-NMR, 13C-NMR, 1H-13C HMBC, IR spectra, and its structure was elucidated. It was identical with 5,8-dihydroxy-3-methyl-3,4-dihydroisocoumarin. At the concentration of 20 mumol/L, its DPPH free?|radical scavenging activity was as 1.67 times as that of vitamin C or 2.10 times as that of Vitamin E. PMID- 12549094 TI - [High concentration ethanol continuous fermentation using yeast flocs]. AB - Continuous ethanol fermentation using yeast flocs was carried out in 4 air-lift suspended-bed bioreactors operated in series. Drafted by CO2, with complete recycle of ethanol distilled effluent broth and at the dilution rate of 0.2/h, the average ethanol concentration of the fermentation broth was 96.6 g/L, while the average concentration of residual total sugar was 4.1 g/L and residual reducing sugar was 1.2 g/L. PMID- 12549095 TI - [Biomass, sporulation and aphid-infecting virulence of Pandora neoaphidis mycelia produced in repeated liquid culture]. AB - Repeated culture of entomophthoraceous isolates is a suspectable factor leading to their virulence decline of biological variation. In the present study, an isolate of Pandora neoaphidis, F98028, was repeatedly cultured six times at the regime of 20 degrees C and 80 r/min in Sabouraud dextrose broth(SDB) at three inoculation ratio of 1/20, 2/20, and 4/20 (seed culture over fresh SDB). Mycelial biomass(MB), sporulation capacity(SC), and virulence to the green peach aphid, Myzus persicae, were assessed for each of the subcultures. The repeated liquid cultures, initiating from seed culture with MB 8.84 mg/mL ad SC 7.22 x 10(5) conidia/mg, yielded MB and SC in a range of 6.4-10.0 mg/mL and 7.3 x 10(5)-10.8 x 10(5) conidia/mg at the ratio of 1/20(72 h culture), 5.7-8.5 mg/mL and 10.0 x 10(5)-12.1 x 10(5) condia/mg at 2/20(60 h culture), and 5.5-10.9 mg/mL and 6.4 x 10(5)-10.9 x 10(5) conidia/mg at 4/20(48 h culture), respectively. There was no significant difference in both MB and SC among the cultures(F = 0.299, P = 0.903) or the three inoculation ratios (F = 0.561, P = 0.587). Inocula from each of the cultures at the inoculation ratio of 1/20 were repeatedly bioassayed on the second and third instar colonies of M. periscae. Based on time-dose-mortality analysis, the estimates of LD50 s for all batches of the inocula were 22.8-162, 4.7-49.4, 2.8-16.7, 2.3-9.5, and 1.8-5.2 conidia/mm2 3-7 d after inoculation, respectively. All the estimates for the repeated cultures fell within a range previously reported. Thus, the method used for repeated liquid culture in this study did not cause a visible decline in the virulence of P. neoaphidis F98028. PMID- 12549096 TI - [Detection of trehalose in transgenic tobacco by HPLC with ELSD]. AB - The E. coli trehase synthalose gene(otsA) was transferred into Nicotiana tabacum mediated by Agrobacterium, but the method for detecting low concentration of trehalose in transgenic plant was not available. The high performance liquid chromatograph(HPLC) with evaporative light-scatting detector(ELSD) using water: methyl cyanide(1:2.6 v/v) as mobile phase was established in this work. An ODS column Zorbax RX-SIL was employed. the trehalose detection limits of ELSD was 5 mg/L. From the linear relationship between the logarithm of trehalose concentration and the logarithm of peak area, it was shown there was 14.7 micrograms.(g FW)-1 in transgenic plant. The data strongly confirmed that trehalose was responsible for the improved stress tolerance of the tobacco. PMID- 12549097 TI - [The new biocontrol strategy of algae pollution]. PMID- 12549098 TI - [Cell permeabilized and it's application]. PMID- 12549099 TI - [Agrobacterium rhizogenes-mediated transformation and the regeneration of transformants in Alhagi pseudalhagi]. AB - The regenerated shoot segments of Alhagi pseudalhagi were sliced and infected with Agrobacterium rhizogenes strain A4. The hairy roots and transformed calli were obtained through selection on hormone free MS medium. The transformants were cultured on MS medium with 2 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 1 mg/L 6-benzylaminopurine (6-BA) to induce calli. 3 mg/L 6-BA and 0.5 mg/L naphthalene acetic acid (NAA) were applied for shoot differentiation. Shoots were planted on MS medium with 2 mg/L indole-3-butyric acid (IBA) and produced roots. Opine analysis proved the integration and expression of T-DNA in over 95% hairy roots, 75% transformed calli and transformed plantlets respectively. The 81% hairy root cells had normal chromosome numbers (2n = 18). The alterations of chromosome number were observed. After one year of subculturing, the regeneration ability of transformants was maintained. PMID- 12549100 TI - [Observation of the biological characterizations of nasopharyngeal epithelial cells by EB virus infection in early phase of immortalization]. AB - The multi-stage cell model of the nasopharyngeal carcinoma development in vitro by Epstein-Barr virus transformation is beneficial for the elucidation of the mechanism of nasopharyngeal cancer. To observe the biological changes of primary human nasopharyngeal epithelial cells in early phase of immortalization, in this study, we have detected the morphological changes and the expression profile of senescence-associated beta-galactosidase (SA-beta-Gal) in primary culture. In addition, the expression of EB virus latent membrane protein 1 (LMP1) and the growth curve of primary cells were also detected. Our results showed a low percentage of cells infected with EB virus expressing SA-beta-Gal activity at the late primary culture. In morphology, the cells also formed multilayer foci, and the cell population doubling time was showed. These results demonstrated that the nasopharyngeal epithelial cells by EB virus infection have passed through the senescence and entered the early phase of immortalization. These cells have some of the transformed characteristics. Our results provided the data for further study on the mechanism of immortalization and the establishment of human nasopharyngeal epithelial cell line. PMID- 12549101 TI - [Mitochondrial permeability transition pore regulates the apoptosis in MGC-803 induced by the extract of glycyrrhiza uralensis Fisch]. AB - In our previous studies, we have discovered that the extract of glycyrrhiza uralensis Fisch (EGUF) can induce obvious apoptosis in gastric cancer cell Line MGC-803. Here, further investigation was carried on about the time-lapse changes of mitochondria transmembrane potential, intracellular free calcium ions, DNA electrophoresis, plasma membrane permeability and chromatin condensation during the apoptotic process of MGC-803 induced by EGUF and the influences of MPT specific inhibitor Cyclosporin A(CsA) on these changes. Enhancement of plasma membrane permeability with PI staining, increase of intracellular free calcium ion and decrease of mitochondria transmembrane potential are early events in apoptotic cascades, prior to the appearances of apoptotic peak, chromatin condensation and DNA ladder. CsA significantly inhibited enhancement of plasma membrane permeability, change of intracellular free calcium ions and decrease of mitochondria transmembrane potential, also greatly delayed the progress of apoptosis. Thus, our results suggest that calcium and CsA-sensitive MPT is involved in the apoptosis of MGC-803 induced by EGUF. PMID- 12549102 TI - [Expression of thyrotropin-releasing hormone receptor mRNA in the rat testis development]. AB - In order to investigate the regulation of thyrotrophin-releasing hormone receptor (TRH-R) expression in rat testis, and to study their function in spermatogenesis, oligonucleotide primers were designed from the sequences of rat pituitary TRH-R cDNA for reverse transcription-polymerase chain reaction (RT-PCR). Specific fragments of TRH-R cDNA were cloned. DNA sequence analysis indicated that cDNA sequence of TRH-R from rat testis was consistent with those of pituitary TRH-R cDNA. The non-radioactive in situ hybridization (NR-ISH) technique was applied to localize cells encoding TRH-R mRNA in the rat testis. Hybridization signals were detected exclusively in the leydig cells, but not in the spermatogenetic cells of the rat testis. TRH-R mRNA in the testis was quantitated in RNA samples prepared from rats at different developmental stages by real time quantitative RT-PCR. The quantitative analyses demonstrated that no TRH mRNA could be detected at the earliest stage (day 8). TRH mRNA signals were detected on day 15 and increased progressively on day 20, 35, 60 and 90. These results suggested that rat testis could specifically express TRH-R, and the transcription of TRH-R gene in the rat testis was development-dependent. PMID- 12549103 TI - [Analysis of genetic diversity of Bama miniature pigs and Guizhou miniature pigs by RAPD]. AB - RAPD analysis was performed with 31 selected single primers to study genetic diversity of two strains of miniature pigs, The percentages of polymorphisic loic in or between two strains of miniature pigs were 30.9%, 29.2% and 25.7% respectively, and the average genetic distances in and between two strains of miniature pigs were 0.120, 0.072, and 0.067 respectively. These results suggested that genetic diversity and genetic variability were poorer in and between two strains of miniature pigs than ordinary breeds' pigs. PMID- 12549104 TI - [Effects of interleukin-2 on the isolated rat heart and the mechanism]. AB - The purpose of the present study was to explore the biological effects and mechanism of interleukin-2 (IL-2) on the isolated rat heart. The results showed that hrIL-2 increased the number of premature ventricular contraction, heart rate, left ventricular developed pressure, left ventricular end-diastolic pressure and coronary flow in the isolated perfused rat heart. Heat inactivated hrIL-2 had no effect on the heart. Pretreatment with ryanodine canceled the positive effects of hrIL-2 on left ventricular developed pressure, left ventricular end-diastolic pressure and coronary flow but had no effects on arrhythmogenesis and tachycardia by hrIL-2. Pretreatment with nifedipine or low extracellular calcium abolished the arrhythmogenic effect of hrIL-2 and attenuated partially the augment of heart rate, left ventricular developed pressure, left ventricular end-diastolic pressure and coronary flow. It suggests that the cardiac activity of hrIL-2 depended on the integrity of its spatial structure and transmembrane influx Ca2+ and intracellularly stored Ca2+ were involved in the cardiac activity of hrIL-2. PMID- 12549105 TI - [The effects of Ag+ on the absorption of trace metal ion during the somatic embryogenesis of Lycium barbarum. L]. AB - We used callus of medical plant, Ningxia Lycium barbarum. L as material to induce somatic embryo genesis, and used multitracer technique and gamma-ray energy spectrum analytical method to study different concentration of AgNO3 on some absorption of trace metal ions during the somatic embryogenesis of Lycium barbarum. L. The results show that (1) when the concentration of AgNO3 is less then 50 mg/L, with the increase of AgNO3 concentration, the absorption rate of some metal ions is increasing, but when the concentration is excess of such concentration, the effects on the absorption rate are different. Ag+ has the effect of coordination, resistance or competition on the absorption of metal ions. (2) AgNO3 in suitable concentration will promote cell differentiation and somatic embryogenesis. When the concentration of AgNO3 is less then 50 mg/L, along with the increase of the concentration, the frequency of somatic embryogenesis is increased, and Ag+ has the promotion on the somatic embryogenesis of Lycium barbarum. L. While the concentration is up to 50 mg/L, it will mostly raise somatic embryogenesis in calli, the amount of callus is about 3 times as much as that of contrast group (in which AgNO3 is not added). When excess of this concentration point, Ag+ has toxic effect, and the somatic embryogenesis is inhibited remarkably. PMID- 12549106 TI - [High-level expression of human calmodulin in E. coli and its effects on cell proliferation]. AB - The gene coding for human CaM was amplified by PCR in which pUC/hCaM3 cDNA was usd as template. After inserting the hCaM III cDNA into the expression plasmid pBV220, we constructed the hCaM3 cDNA-recombinant expression vector(hCaM3/pBV220). The recombinant plasmid was then transformed into E. coli DH5 alpha. After heat induction, a high level expression of CaM protein was obtained. SDS-PAGE analysis showed that the recombinant E. coli could express a 17 kD protein which accounted for about 20% of the total cellular protein. Western blot analysis showed that anti-CaM monoclonal antibody(McAb) specifically bound to the 17 kD band of expression product. rhCaM was purified by Phenyl sepharose CL-4B affinity chromatography from recombinant bacterial lysate. 3-4 mg of the purified protein were obtained from 1 liter of bacterial culture. The rhCaM was able to activate NAD kinase to the same extent as the standard human brain CaM (Sigma). K562 cells and SP2/0 cells were seeded in 24-well or 96-well plate and cultured for 48 h with rhCaM and CaM-antagonist trifluoperazine(TFP). Cell proliferation rates was determined by MTT assay. There was a significant positive correlation between the concentrations of rhCaM and the cell proliferation rates. CaM-antagonist TFP had an inhibitory effect on cell proliferation rate. The inhibition could be corrected by the addition of extracellular rhCaM. PMID- 12549107 TI - [Somatic embryogenesis and plant regeneration in vitro from young shoots of Aralia elate (Miq.) Seem]. AB - Explants excised from the young shoots of Aralia elata (Miq.) Seem. were cultured on MS media. Calli were induced from the explants on MS medium supplemented with 0.5 mg/L 2, 4-D, 0.5 mg/L BA and 0.5 mg/L NAA. Then these calli were transferred onto the MS medium containing 2.0 mg/L 2,4-D + 0.5 mg/L BA + 0.5 mg/L NAA and 0.2% activated charcoal. Under these conditions the somatic embryoids were observed and regenerated plants were obtained from somatic embryogenesis. Then, a experimental system with stability and high regenerating efficiency has been set up for the propagation of the young plants, the cell breeding technology and the control of somatic embryogenesis of Aralia elata (Miq.). PMID- 12549108 TI - [Cloning and efficient expression of cytokine human MK in E. coli]. AB - For cloning the cytokine human Midkine (MK) gene, we designed by PCgene program and synthesized a pair of PCR specific primers according to the reported human MK cDNA sequence. Total cellular RNA was extracted from a human hepatoblastoma cell line HepG2, and then the target DNA fragment was obtained by RT-PCR and subcloned into plasmid pUC118. Checked with radioisotope sequencing and ABI 377A sequencer, the nucleotide sequence of the cloned MK cDNA was identical with the reported one. A prokaryotic expression vector, named pBV220, was used to express the MK protein efficiently in E. coli strain TG1 and a predicted band of 16.5 kD in Mr by 15% SDS-PAGE was found. The expressed recombinant protein was found in insoluble aggregated form and accounted for about 31.21% of the total cellular proteins. The first 15 N-terminal amino acid sequence analysis of this protein by Edman degradation method showed that it was accordant with that predicted from the cDNA sequence. The activity of neurite outgrowth-promoting of the MK crude samples was tested with brain cells isolated from 18-day embryos of SD rat. PMID- 12549109 TI - [Molecular cloning of full length capsanthin/capsorubin synthase homologous gene from orange (Citrus sinensis)]. AB - The complete sequence of orange homologous capsanthin/capsorubin synthase gene is 3788 bp long with a coding sequence of 1512 bp, which encodes a polypeptide of 503 amino acids. The 5' upstream sequence is 1721 bp long and the 3' downstream sequence is 555 bp long. The amino acid sequence of this gene is 78% and 69% identical to the genes from carrot and pepper, respectively. It is also partially homologous to plant neoxanthin synthase, lycopene beta-cyclase and lycopene epsilon cyclase genes. Isolation of the gene provides a framework for elucidation of the mechanisms involved in inability of citrus to produce capsanthin and capsorubin. PMID- 12549111 TI - [Reclamation of the plant regeneration efficiency of Brassica campestris subsp. chinensis var.Parachinensis]. AB - This investigation has developed an efficient and fast method for plant regeneration from petiole of cotyledon explants of Brassica campestris L. subsp. chinensis Makino var. parachinensis Tsen et Lee. A medium was designed for B. campestris subsp. chinensis var. parachinensis to obtain the high frequency of shoot regeneration, which contained BAP 2 mg/L, NAA 0.75-1.0 mgL and 7.5 mg/L AgNO3 solution to the half of NH4+ concentration's MS basic medium. 60 mL/L coconut milk were added to all of media. In this method, frequency of shoot regeneration of "youqing caixin" reached as high as 91.2% and the number of shoots per explant reached as high as 4.7 plants. The result showed that there was a positive correlation between frequency of shoot regeneration and number of shoots per explant. The little shoots could be observed five days after inoculation and were formed directly. The inducing rate of roots of the shoots reached as high as 100% and the rate of viability of transferred mature plant reached higher than 95%. The regeneration period from petiole with cotyledon to a seedling was shorten to about 49 days. Factors influencing in vitro explant regeneration were studied. PMID- 12549110 TI - [Genetic instability in cancer tissues analyzed by RAPD PCR]. AB - In five kinds of tumors, total 128 specimens were analyzed by RAPD (random amplified polymorphic DNA) PCR with nine 10-base arbitrary primers for detecting instabilities of DNA and chromosome and screening new molecular markers coupled to putative or unknown oncogenes and/or tumor suppressor genes. Bands representing instabilities have been recovered and purified from agarose and cloned into pCAPs vector, and further labeled by DIG as probes for analysis of Southern blot, Northern blot and Sequencing. Results revealed that sample 5 and 3 of the gastric cancers showed the highest genomic changes and the average detectability in five sorts of cancers was up to at least 40% (42.2%-49.4%), and that there were significant differences in the ability of each primer to detect genomic instability, which ranged from 27% to 68%. Despite the highest detectability of genetic instability (68%) in tumor tissues, primer 2 could produce stable profiles of DNA bands in normal tissue genome with good reproducibility. On the contrary, primer 8 was of the lowest one (27%). Band B of single copy found to be allelic losses in gastric and colon cancers according to RFLP analysis was of a novel sequence and registered by Gen-Bank (Accession Number AF151005). Therefore the genetic instabilities often concentrated on some special locuses of chromosome e.g. repetitive sequences etc. and coupled to carcinogenesis. It was impossible or difficult to get great achievements for cancer treatments with the procedure of gene therapy only to one oncogene or one tumor suppressor gene because the extensive DNA variations occurred during the progression of tumor. RAPD assay connected with other techniques was a good tool for the detection of genomic instabilities and direct screening of some new molecular markers related to tumor suppressor genes or oncogenes. PMID- 12549112 TI - [Study on the steroid constituents of soft coral Lobophytum microspiculatum]. AB - The soft coral Lobophytum microspiculatum collected from Xisa Islands of the South China Sea was immersed in and extracted with ethyl alcohol and the extract was concentrated, and then partitioned between ethyl acetate and water. The portion soluble in ethyl acetate was subjected to column chromatography over Sillca H, eluted with a step gradient of EtOAc and petroleum ether. The sterols were obtained from 20% EtOAc-petroleum ether eluent portion. By means of GC/MS method, the sterols were identified to be: cholesta-26-methyl-ol; ergosta-8, 22 dien-3 beta-ol, 24-methyl; ergosta-5, 22-dien-3 beta-ol, 23, 24-dimethyl; cholesta-5,22(23)-dien-3 beta-ol, 22,24-dimethyl; cholesterol; ergosta-5,22-dien 3 beta-ol, 24-methyl; ergosta-5,22-dien-3 beta-ol, 22,24-dimethyl, 24-methyl cholesterol; ergosta-7-en-3 beta-ol, 24-methyl; nor-ergosta-7-en-3 beta-ol and gorgosterol. Among them cholest-5,22(23)-3 beta-ol, 22,24-dimethyl (4) is a new compound which was found at the first time from the nature. This finding gave a further prove for the biogenesis path way of gorgosterol. PMID- 12549113 TI - [Retention behavior of solutes on liquid chromatographic column packed with dynamically modified zirconia]. AB - Zirconia was dynamically modified with stearic acid, beta-cyclodextrin and sodium dodecylsulfonate, separately, to form liquid chromatographic packings. The chromatographic behaviors of aromatic hydrocarbons, acidic compounds (phenol, nitrophenols and benzoic acids) and basic compounds (aniline, nitroanilines and toluidines) were investigated on these modified zirconia packings with a mixture of methanol/water as the mobile phase. On stearic acid modified zirconia, aromatic hydrocarbons and the basic compounds exhibit symmetrical peaks. The retention time of these solutes decreases with increasing the concentration of methanol in the mobile phase. When the mobile phases with stearic acid were used, the retention time of these solutes has a maximum with variation of stearic acid concentration (0-1.0 mmol/L) in the mobile phase. However, the acidic compounds such as nitrophenols and benzoic acids are strongly adsorbed and can not be eluted from the column with the mobile phases. No leaking of stearic acid on the modified zirconia was found after 3000 column volumes of the methanol/water mobile phase were used. The results showed that the stearic acid modified zirconia can be used as a stationary phase for reversed-phase liquid chromatography. On beta-cyclodextrin modified zirconia, aromatic hydrocarbons and basic compounds exhibit shorter retention time than those on the stearic acid modified zirconia under the same mobile phase composition, however, nitrophenols can be eluted but give unsymmetrical peaks. On sodium dodecylsulfonate modified zirconia packings, the basic compounds can not be retarded; Aromatic hydrocarbons and acidic compounds exhibit short retention time. The results can be ascribed to the weak adsorption of sodium dodecylsulfonate on the zirconia. PMID- 12549114 TI - [The study of vapor-liquid equilibria for polymer/solvents by using gas-liquid chromatography]. AB - The infinite dilution weight fraction activity coefficient, omega 1 infinity, and Flory-Huggins interaction parameter, chi, have been determined for systems of polydimethylsiloxane (PDMS)/solvents and polymethyl-methacrylate (PMMA)/solvents by using gas chromatography from 58 degrees C to 180 degrees C. The solvents include styrene, acetone, ethyl ether, methanol, ethanol, acetic acid, trichloromethane, tetrahydrofuran and ethyl acetate in PDMS/solvent systems and n hexane, n-heptane, cyclohexane, benzene, toluene, ethyl benzene, styrene, acetone, ethyl ether, methanol, ethanol, methyl methacrylate(MMA), trichloromethane, tetrahydrofuran and ethyl acetate in the PMMA/solvent systems. The results showed that omega 1 infinity and chi were decreased with the increase of temperature in the range from 58 degrees C to 180 degrees C. The study showed that UNIFAC and UNIFAC-FV models could not all be used to estimate omega 1 infinity of the solvents in PMMA/solvent systems. Therefore, UNIFAC and UNIFAC-FV models must be corrected or other models must be used. PMID- 12549115 TI - [Separation of alkylphenols in oil field water by micellar electrokinetic chromatography]. AB - The abundance of alkylphenols in the system of water, oil and rock is of great geochemistry significance. The analysis of alkylphenols in oil field water using micellar electrokinetic chromatography with UV detection is described. The different variables that affect separation were studied and optimized. The results show that better separation was obtained with the borate concentration of 47.5 mmol/L, the SDS concentration of 70.0 mmol/L, pH 9.15, separation current of 100 microA, capillary temperature 20 degrees C. The organic additive(beta-CD) could partly improve separation. Water sample was introduced into the capillary by pressure of 5 kPa for several seconds. Detection wavelength was 200 nm and reference wavelength at 350 nm for all experiments. For the oil field water sample, the reproducibility of directly injected oil field water after filtration through a membrane filter of 0.45 micron pore size was far from good and it's concentrations of alkylphenols were too low to be detected. Therefore, a method of separation and enrichment by a column packed with tributyl phosphate(TBP) extraction resin was applied for analysis of TZ4 oil field water. Qualitative analysis show that there are phenol, o-cresol, m-cresol and p-cresol in this water sample and phenol is the main component and it's quantity is more than other alkylphenols. It's recoveries from the oil field water were 91.5%-95.3%. The results were satisfactory. PMID- 12549116 TI - Determination of selenium compounds by HPLC with ICP-MS or FAAS as selenium specific detector. AB - A speciation method was developed for selenious acid, selenic acid, trimethylselenonium ion (TMSe) and selenomethionine (SeMet). Separation of the four selenium species was achieved by HPLC on an ESA Anion III anion-exchange column using aqueous mobile phase of 5.5 mmol/L ammonium citrate at pH 5.5 with a flow rate of 1.5 mL/min. Under the optimal conditions, the four selenium species were separated within 8 minutes. On-line selenium-specific detection was carried out with an inductively coupled plasma mass spectrometer (ICP-MS) or a flame atomic absorption spectrometer (FAAS). The detection limits of HPLC-FAAS were approximately rho(Se) = 1 mg/L for each compound (100 microL injection). To increase the nebulization efficiency of the ICP-MS, the Meinhard concentric nebulizer was replaced by an ultrasonic nebulizer (USN). The ICP-MS signal intensity was increased by a factor of 7 for selenious acid and 24 to 31 for TMSe, SeMet and selenic acid with the USN compared to that with the Meinhard nebulizer. The detection limits of the HPLC-USN-ICP-MS were rho(Se) = 0.08 microgram/L for TMSe, rho(Se) = 0.34 microgram/L for selenious acid, rho(Se) = 0.18 microgram/L for SeMet and rho(Se) = 0.07 microgram/L for selenic acid. PMID- 12549117 TI - Method development of adsorbed stationary phase open tubular capillary electrochromatography. AB - The adsorption effect of capillary wall, which was usually considered as a troublesome factor, has been used as driving force for preparing stationary phase for open tubular capillary electrochromatography (OTCEC). A number of compounds have been applied as stationary phase materials, which include cationic surfactants such as cetyltrimethylammonium bromide (CTAB), basic proteins such as lysozyme and cytochrome C, basic peptides such as Lys-Tyr and Lys-Ser-Tyr, and basic amino acid L-lysine. The adsorbed CTAB phase is used for separation of neutral compounds while other adsorbed stationary phases are used for chiral separation. The run-to-run reproducibility of retention time is rather good with RSD values less than 2.3%. The separation efficiency is excellent with the highest theoretical plate number of up to 590,000/m and the average plate number of more than 250,000/m. Being stored at 2-8 degrees C in refrigerator, the adsorbed stationary phase can last at least one month. PMID- 12549118 TI - Supercritical fluid CO2 extraction of colchicine from colchicine Autumnele liliaceae. AB - Colchicine was successfully extracted from the corms of colchicine autumnele Liliaceae using supercritical fluid (SF) CO2. The pressure range used in the experiments was 20-40 MPa and the temperature was 40 degrees C. Being its high molecular polar, colchicine or other alkaloids are not easy to be extracted by SF CO2. In order to increase the mass transfer from the sample matrix to SF, different presoakers (such as methanol, ethanol, acetone etc.) were employed. Among the selected pre-soakers, best results were obtained with ethanol both in the aspects of yield and selectivity. At 35 MPa, 40 degrees C and with ethanol as pre-soaker, the yield was 1.8 mg/g, and the colchicine could be almost completely extracted from the sample matrix. The average concentration increased from 0.18% in natural sample to 6.92% in extract with SFE processing only once under the given conditions. The comparison of liquid ethanol extraction (LEE) and SFE showed that SFE is greatly superior to LEE in colchicine separation with SFE not only higher yield and selectivity but also a quick, safe and non-toxic method were obtained in colchicine extraction. PMID- 12549119 TI - [Advances in affinity chromatography]. AB - This review is mainly concerned with the recent advances in (i) mechanism of affinity chromatography; (ii) the use of new techniques such as combinational chemistry, molecular imprinting, de novo design, fusion expression in discovery of ligand; (iii) improvement of support and chromatographic system; (iv) application of affinity chromatography. The review includes 66 references. PMID- 12549120 TI - [Molten organic salt--a new class of stationary phases for GC]. AB - Molten organic salts are a new class of polar, selective stationary phases for gas chromatography. Their types and characters are reported in detail and compared with some common stationary phases. PMID- 12549121 TI - [Analysis of C5 fractions and carbonyl compounds by capillary gas chromatography]. AB - We used two capillary columns with different characters--SE-54 (30 m x 0.25 mm) and Squalane (92 m x 0.24 mm) to analyse C5 fractions and carbonyl compounds in the mixtures of complex chemical composition. The physical properties of some components are very similar, especially for 1,3-cyclopentadiene and cis-1,3 pentadiene. 1,3-Cyclopentadiene in the feed will directly affect the activity of catalyst. In the past we only used SE-54 column to separate 1,3-cyclopentadiene and cis-1,3-pentadiene, but the results were unsatisfactory. Later we found that squalane column could separate them fairly good, but it could not yet separate dicyclopentadiene from 1,3-cyclopentadiene and carbonyl compounds in C5 fractions very well. In this work SE-54 capillary column and squalane capillary column were used together to analyse C5 fractions, and SE-54 column was used to analyse carbonyl compounds. The results are satisfactory. PMID- 12549122 TI - [Detection of 8-hydroxyguanine in the DNA of the H2O2-treated HL-60 cells by GC/FID]. AB - This paper describes the DNA damage by hydrogen peroxide in HL-60 cells and the detection of 8-hydroxyguanine produced. The number of cells was at least about 2 x 10(7)/sample. Treatment of HL-60 cells with 0.4 mmol/L hydrogen peroxide for 24 h would result in DNA damage. The DNA samples were isolated, then qualitatively and quantitatively analyzed before hydrolysis. The results of experiments showed that 100-300 microL 98% formic acid could hydrolyze about 0.12 mg DNA samples completely in 150 degrees C for 1 h. After lyophilizing the DNA samples for 6-8 h, one DNA sample was trimethylsilylated completely with 150 microL BSTFA in 150 degrees C for 1 h. The recovery of trimethylsilylated 8-hydroxyguanine was 81.70%. The trimethylsilylated 8-hydroxyguanine was identified by GC/FID and CGC/MS-SIM parallely. PMID- 12549123 TI - [Determination of triptolide in tripterygium preparations by gradient high performance liquid chromatography]. AB - A gradient elution method was developed for the separation and determination of triptolide in Tripterygium wilfordii Hook preparations by high performance liquid chromatography (HPLC) using YWG column made in China. Methanol with 0.05 mol/L KH2PO4 was used as mobile phase, and the photodiode array detector was used at lambda = 218 nm. The sample of tripterygium oral preparation was extracted with ethanol and then by chloroform. The extract solution was evaporated and the residue was dissolved in 10 mL of ethanol as a sample solution. The spectrum was obtained by the subtraction of spectrum of ethanol from that of sample solution. In comparing with isocratic elution, the gradient operation used in this paper had higher resolution and sensitivity. The calibration curve was linear over the range of 1.32-21.10 mg/L (r = 0.9999). The average recovery of the method was 97.59%. The method was applied to separate and determine the triptolide in tripterygium tablets and the results indicated that the triptolide content by this method was less than that by isocratic and label-claimed one. PMID- 12549124 TI - [Simultaneous determination of 5-component contents in Kuaian anticold solution by high performance liquid chromatography]. AB - A reversed-phase, isocratic high performance liquid chromatographic method has been successfully used to separate and determine the 5 components simultaneously in Kuaian anticold solution. In the anticold solution, paracetamol, caffeine, DL methylephedrine HCl, chlorpheniramine maleate and guaiacol glyceryl ether were analyzed on a NOVA-PAK C18 column (150 mm x 3.9 mm i.d., 5 microns) at 35 degrees C with V(acetonitrile):V(0.03 mol/L ammonium dihydrogen phosphate):V(0.004 mol/L sodium heptyl sulfonate):V(triethylamine) = 12:44:43.5:0.5 with pH adjusted to 3.0 by orthophosphoric acid as the mobile phase. The detection was performed at 220 nm and the injection volume was 15 microL. For the 5 components mentioned above the calibration curves were linear in a certain range. The recoveries were (99.6 +/- 1.3)%, (98.4 +/- 1.7)%, (98.0 +/- 1.5)%, (98.5 +/- 1.8)%, (100.3 +/- 1.7)% (n = 5) respectively. PMID- 12549125 TI - [Study on high performance liquid chromatography/indirect photometric detection of fosfomycin with acridine as detection reagent]. AB - Fosfomycin is a broad-spectrum antibacterial agent which has strong polarity and weak UV absorption. In this work, a method using standard ODS column and conventional UV detector set at 254 nm to analyse fosfomycin was investigated. Firstly, several UV detection reagents were tested and acridine(A) was found to give the highest indirect detection response and proper modification on retention of fosfomycin. With acridine as the UV detection reagent, the mobile phase parameters which affect retention and detection response of solute are discussed, e.g. pH, concentration of organic solvent and concentration of acridine. The mechanism is interpreted for each case. After optimization, we adopted mobile phase as V(CH3OH):V(H2O) = 50:50, pH 3.18, [HAc] = 2 mmol/L, [A] = 1 x 10(-5) mol/L. A new chromatographic system monitored by indirect photometry for detection of fosfomycin was established. This method was used for quantitative analysis, the minimum detectable level was 10 mumol/L, the relative deviation was 4.51%. PMID- 12549126 TI - [Separation of p-aminobenzenearsonic acid and its oxide by ion-pair reversed phase high performance liquid chromatography]. AB - To study the separation of p-aminobenzenearsonic acid (PABAA) and its oxide, p aminophenylarsine oxide (PAPAO), both the absorption spectra were scanned at the wavelengths from 200 nm to 380 nm. PABAA had absorption maximum at 254 nm and PAPAO 258 nm. The effects of salt concentration, column temperature, methanol and ion-pair agent concentrations on the capacity factor were investigated. Compounds of high polarity showed almost no retention on reversed-phase column; as the volume fraction of the methanol decreased from 90% to 10%, the retention time of PABAA gradually increased with broad peak, and partially eluted when methanol volume fraction being below 20%. With temperature rising, the retention time of PABAA was decreased. But PABAA capacity factor can be increased by selecting an appropriate salt concentration for the mobile phase. The cetyltrimethyl and tetrabutyl ammonium ions were separately added as ion-pair agents to the mobile phase containing methanol in phosphate buffer of 10 mmol/L, the changes of retention time were observed. The mechanism of retention based on reversed phase ion-pair model is proposed. Besides, the retention behaviour is also influenced by size exclusion in stationary phase as well as polar interactions with residual silanol group on the silica surface. PMID- 12549127 TI - [A rapid separation and quantitation of sodium cyclamate in food by ion-pair reversed-phase high performance liquid chromatography]. AB - A method of separation and quantitation of sodium cyclamate in foods by ion-pair reversed-phase high performance liquid chromatography is described. Sodium cyclamate can be separated on ODS column at room temperature by using a mobile phase of V(methanol):V(water, containing ion-pair reagent) = 30:70 at 1 mL/min flow rate. The effects of ion-pair reagent and methanol on the retention of sodium cyclamate were studied. The samples were quantified with an ultraviolet detector operated at 205 nm. The calibration curve was linear in the range of 0.50-2.50 g/L with a correlation coefficient of 0.9996. The recoveries were 96.9% 101.7%. The detection limit was 0.05 g/L. This method is simple, rapid and sensitive. PMID- 12549128 TI - [A novel solid phase coating of solid-phase microextraction prepared with sol-gel technology and its applications]. AB - In this paper, the recent development and applications of solid-phase microextraction (SPME) are summed up. A sol-gel method was applied for the preparation of solid phase coating in SPME. The exposed outer surface was coated with hydroxy-terminated poly (dimethylsiloxane). The characteristics of this solid phase were evaluated. The coating possesses higher thermal stability, and shorter extraction and desorption time. This solid phase appears to have strong affinity to both polar compounds and non-polar compounds. The scanning electron micrograph of the coated surface presents a porous structure. In the analyses of the environmental samples, ideal results were also obtained. PMID- 12549129 TI - [Preparation of zirconia for high performance liquid chromatography using sol-gel technology]. AB - Zirconia sol was prepared by dissolving zirconyl chloride in water, and the sol formed gel in water/cyclohexane W/O style emulsion to obtain monodispersed particles of ZrO2(2-10 microns). After being washed and classified, the zirconia particles (3-5 microns) were collected and calcined at 800 degrees C to remove the organic residues on the surface of the particles. The mean pore diameter, specific surface area and specific pore volume of the zirconia particles were found to be 14.2 nm, 11.2 m2/g, and 0.04 cm3/g, respectively, by means of the BET method. The adsorption behavior of various organic compounds on the ZrO2 shows it will be useful as a column packing material for high performance liquid chromatography. PMID- 12549130 TI - [The determination of anions of very weak acids by incompletely suppressed conductometric detection]. AB - A novel method, incompletely suppressed conductometric detection, has been developed for the determination of anions of very weak acids. The principle of the method is discussed. The advantages of the method over the suppressed and nonsupressed methods for the determination of anions of very weak acids are also discussed theoretically. Results showed that the detection limits of the method for arsenous acid and boric acid were lower than those of nonsuppressed conductometric method. Being compared with suppressed conductometric method, the sensitivity was improved greatly. For 250 mg/L boric acid, 261- and 1002-fold increases in peak height and peak area were obtained, respectively. Within the linear range of the detection, the linear correlation coefficients of the peak areas of two weak acids were 0.9991 and 0.9985, respectively. PMID- 12549131 TI - [Determination of inosine 5'-monophosphate and guanosine 5'-monophosphate in taste-enhancers by ion chromatography]. AB - A high performance anion-exchange chromatographic method was developed for the simultaneous determination of two kinds of taste-enhancers, inosine 5' monophosphate (IMP) and guanosine 5'-monophosphate (GMP), in the presence of monosodium glutamate and chloride (salt). The separation was achieved by using Dionex IonPac AG11 and IonPac AS11 columns, and the determination by ultraviolet absorbance detection at 254 nm. The detection limits (S/N = 3) were 0.21 mg/L and 0.27 mg/L for IMP and GMP, respectively. Good linearities between the concentrations of the two analytes and relevant peak area responses were obtained in the range of 2-100 mg/L. Under the experimental conditions, glutamate, chloride, succinate, benzoate, sorbic acid and citrate did not interfere in the determination of IMP and GMP. The method has been successfully applied to the determination of IMP and GMP in gourmet powder as well as chicken bouillon, and the average recoveries for various samples ranged from 97% to 103%. PMID- 12549132 TI - [Study on chromatographic peak identification by means of peak shape analysis]. AB - A new method for chromatographic peak identification by means of peak shape analysis is presented in this paper. Directly measurable parameters, i.e. peak width and asymmetry factor are first utilized for peak shape analysis. It is showed from theoretical and experimental results that for a component having almost the same retention time with the standard, an overlapped peak can be obtained by injecting the mixture of analyte and standard. In comparing the peak shape of overlapped peak with the standard one, the component can be identified whether it is the same with the standard or not. PMID- 12549133 TI - [Preparation and application of a new chloromethylstyrene encapsulated silica single anion column]. AB - The silanized silica was firstly encapsulated with chloromethylstyrene and then quaternized by N, N-dimethylbenzylamine. Thus a new kind of strong anion stationary phase was prepared. This strong anion silica column has superior separation ability for the anions by using indirect ultraviolet detection with wavelength at 254 nm and the mobile phase of V(methanol):V(0.125 mol/L NaH2PO4) = 60:40. Moreover, separation of mono- and di-petroleum sulfonates in Yumen sample could also be good with gradient elution. The sample was first eluted with 100% A [V(methanol):V(water) = 60:40] for 1 min, then the eluent was changed linearly to 100% B [V(0.125 mol/L sodium phosphate monobasic):V(methanol) = 60:40] within 1 min, then the sample was continuously eluted with 100% B for 6 min. PMID- 12549134 TI - [Determination of triflumizole and its metabolite residues in vegetables by high performance liquid chromatography]. AB - Triflumizole (TRI) and its metabolite [4-chloro-alpha,alpha,alpha-trifluoro-N-(1 amino-2-propoxythylidene)- O-toluidine] (MET) were extracted with methanol and reverse-extracted with dichloromethane. After evaporation to dryness, the residue in dichloromethane layer was dissolved in mobile phase for high performance liquid chromatographic determination with external standard method. The total residue content of TRI and MET (calculated as TRI) is reported. PMID- 12549135 TI - [Determination of 10-HDA in honeybee body by HPLC]. AB - In the present work we found that in the honeybee body there exists an unsaturated fatty acid, trans-10-hydroxy-2-decenoic acid (10-HDA), which was known only to be present in royal jelly. We established the analytical method of 10-HDA in honeybee body by HPLC and simplified the extraction method of 10-HDA. In the optimum conditions the linear range of detection was 10-1,000 ng, the correlation coefficient was 0.9998, the recovery was 96.5%-99.2% and the detectable limit was 0.53 microgram/g. PMID- 12549136 TI - [Simultaneous analysis of alkali metals and alkaline earth metals in Chinese herbal medicine by ion chromatography]. AB - A simple and rapid method for the analysis of cations by using ion chromatography was developed. The results showed that the content ratio of Na+, NH4+, K+, Mg2+, Ca2+ in rhizoma chuanxiong and semen zizyphi spinosae are different. The detection limits of cations (S/N = 3) were 0.001-0.013 mg/L. The calibration curves of peak areas for all analytes were linear over two or three orders of magnitude with a regression coefficient (r2) of 0.9995-1.0000. PMID- 12549137 TI - [Determination of nervonic acid in oral tonic by GC]. AB - A method for determination of nervonic acid in oral tonic has been developed by gas chromatography. The samples were derivatized by BF3-CH3OH. Dinonyl phthalate was used as the internal standard. The calibration curve was linear over a range from 0.3 g/L to 3 g/L with a correlation coefficient of 0.9994. The average recovery of the method was 97.04% and the RSD was 0.90%. The method has been successfully applied to quality control of nervonic acid in oral tonic. PMID- 12549138 TI - [Determination of nifedipine content and its uniformity in tablets by high performance liquid chromatography]. AB - An HPLC method with diazepam as internal standard was developed to determine nifedipine content and its uniformity in tablets. The analytical column was a C18 column, the mobile phase consisted of V (methanol):V(triethylamine, pH 5.20) = 68:32. The detection was carried out with UV detector (lambda = 237 nm). Under this condition, the linear range was wide(8-80 mg/L) and the recovery was high (99.86%). The proposed method is simple and accurate and the decomposed product can be separated as well. Therefore, the quality of nifedipine tablets can be effectively controlled. PMID- 12549139 TI - [Determination of plasma homocysteine in normal adults by high performance liquid chromatography with fluorescence detection]. AB - A rapid, isocratic high performance liquid chromatographic(HPLC) method with fluorescence detection was used for the determination of total homocysteine (Hcy) levels in human plasma. Prior to reversed-phase HPLC analysis, the plasma thiols were derivatized with ABDF(7-fluoroben30-2-oxa-1,3-diazole-4-sulfonamide), a thiol specific fluorogenic probe. A mobile phase consisted of 0.1 mol/L potassium dihydrogen phosphate (pH 2.1, containing 10% of acetonitrile) ensured base line separation of plasma Hcy. The mean recovery of this method was 97.75%. The intraday and interday coefficient of variations were 4.49% and 9.79%, respectively. The method is simple, sensitive, reproducible and very suitable for routine determination of plasma Hcy in human. PMID- 12549140 TI - [The determination of F-, Cl-, NO3- and SO(4)2- in the total suspended particles in air by ion chromatography]. AB - In this paper the method used for the determination of F-, Cl-, No3- and SO(4)2- in the total suspended particles in air (TSP) by ion chromatography was established. The contrast between the blank values of filter membranes of ultra thin glass fiber and chlorinated PVC, the contrast between extraction methods by supersonic extraction with 70 degrees C water and by just stand for 24 h, and the contrast between extraction solutions with deionized water and the eluent (2.2 mmol/L Na2CO3-2.8 mmol/L NaHCO3) were involved. The determination of the uniformity of filter membrane and the typical samples were also carried out. This paper adopted ethylene tetrachloride filter membrane, deionized water and supersonic for extraction. The average recoveries of this method for F-, Cl-, NO3 and SO(4)2- were 98.7%, 103%, 101% and 99.8% respectively and the RSD's were 0.29%, 0.32%, 0.30% and 0.25% respectively. This method possesses better recovery and precision. PMID- 12549141 TI - [Discussion on the concept problem of the specific retention volume in gas chromatography]. AB - The concept problem in the definition of well-known specific retention volume in GC is proposed in this paper. The specific retention volume Vg is usually defined as the net retention volume per gram of liquid phase corrected to 273.15 K in some literatures, and is widely appeared in journals and textbooks as one of the most important absolute retention value in GC, though it is less practical than the relative retention value in the qualitative analysis. Being inspired in the discussion of pressure correction in GC, the author thinks that the temperature correction term in the Vg definition is neither correct nor necessary. The problem was originated from the misunderstanding of the pressure correction of the adjusted retention volume. It is emphasized here that the net retention volume obtained by correction of the adjusted retention volume by compressibility factor j is the retention parameter in term of the column geometrical volume rather than a gas volume. The net retention volume per gram of liquid phase is an absolute retention parameter sufficient to measure the retention power. The author thinks that it is necessary to discuss and clarify the problem to avoid any possible errors being introduced in some cases, such as in the estimation of some thermodynamic parameters. PMID- 12549142 TI - [Determination of alpha-methoxynaphthalene by reversed-phase high performance liquid chromatographic method]. AB - A reversed-phase high performance liquid chromatographic method was developed for the quantitative determination of alpha-methoxynaphthalene and it's harmful impurity. The instrument used was Varian Model 5000 HPLC system, with Nucleosil C18 column, 10 microns, 300 mm x 4.6 mm i.d., eluent of V (methanol):V(water) = 90:10 at a flow rate of 0.8 mL/min and detection wavelength, 254 nm. The relative purity of alpha-methoxynaphthalene was more than 96% and relative standard deviation was 0.29%. PMID- 12549143 TI - Two approaches for determining the urinary excretion patterns of nucleosides- HPLC and CE. AB - Cancer patients excrete in their urine increased amounts of modified purines and pyrimidines. These modified mucleosides are primary constituents of RNA and are synthesized at the macromolecular level. When RNA is catabolized, most of these modified nucleosides cannot be reutilized; consequently, they are excreted. Due to their increased urinary excretion in conjunction with altered RNA turnover in carcinogenesis, they have been proposed as tumour markers. We developed both reversed-phase high performance liquid chromatographic and capillary electrophoretic approaches to determine normal and modified nucleosides in urine. Data obtained with CE are in good accordance with those from HPLC. The reference excretion levels of urinary nucleosides from healthy volunteers were established, and elevated ones of modified nucleosides from 34 patients with different kinds of cancer were observed. The developed methods are suitable for the analysis of large series of samples for clinical studies. PMID- 12549144 TI - [The effect of mobile phase on the enantiomeric separation of cyfluthrin by chiral high performance liquid chromatography]. AB - The enantiomeric separation of cyfluthrin on chiral stationary phase(CSP), derived from (R)-N-(3,5-dinitrobenzoyl) phenylglycine (Prikle type 1-A CSP) has been studied by using different hexane-alcohol mixtures as mobile phase. Mobile phase strength and composition have shown an important role in the chromatographic separation of eight isomers of cyfluthrin. Increase of the mobile phase strength reduces the retention of the eight isomers, while the diastereomeric selectivity is decreased and the enantiomeric selectivity remains almost unchanged. Also, the resolution varies in accordance with the steric hindrance of the alcohol. Secondary or tertiary alcohols impart a greater resultion to the CSP than primary alcohols. Peak shapes are better with the lower alcohols than the higher alcohols. For the enantiomeric separation of cyfluthrin on Pirkle type 1-A CSP, hexane-tert-butanol and hexane-2-propanol are better binary mobile phases. PMID- 12549145 TI - [Protein A tangential flow membrane chromatographic column used for immunoadsorption therapy]. AB - Tangential flow membrane chromatographic column is a new model of immunoadsorption therapy from blood. The results showed that the structure of the column and the flowing channel had a little damages for blood cells. The relation of flow-rate and backpressure with water, plasma and blood individually as the fluid flowing through the column has been studied. It was observed that backpressures increased with fluid viscosity, when flow-rate of blood was 120 mL/min and the backpressure reached 0.093 MPa; The adsorption capacity of protein A of the column for IgG from human plasma has been measured. When plasma and blood individually circulated through the column for 1 h, the column with 139 mg protein A immobilized on the matrix (6 mg protein A/g dry matrix) adsorbed 553 mg IgG (23.8 mg IgG/g dry matrix) from human plasma and 499.4 mg IgG (21.5 mg IgG/g dry matrix) from human blood respectively. The circulation time had a great influence on IgG adsorption capacity; but the circulation rate had a little influence on IgG adsorption capacity. The result of extracoporeal blood perfusion for dog showed that the Protein A column has good blood biocompatibility. PMID- 12549147 TI - [Development of column switching systems in industrial gas chromatography]. AB - According to the home-made requirement of column switching systems in 23 industrial gas chromatographs in 300,000 Tons of olefin plant, based on the research work of the expert system on gas chromatography and movement laws of compounds of interest in columns, the suitable stationary phases were defined, and the column switching systems were designed by using combined cutting techniques which consist of backflushing, fore-flushing and heart cutting, and by controlling column lengths, flows, flow directions and temperatures. The practical applications showed the column systems satisfy the on-line monitoring requirement of the production process and can be used continually for 1-3 years. PMID- 12549146 TI - [The application of artificial neural network in QSAR research of PCDD]. AB - The study of QSAR between molecular structure and retention behavior of solutes is an important topic of chromatographic basic theory. PCDD (polychlorinated dioxin) is a kind of important environmental toxin. The basic skeleton of PCDD is [symbol: see text]. PCDDs are all chlorine substituted, so they are only different from each other in the number and position of chlorine substitutents on the skeleton. There are 76 kinds of homologues (1 to 8 chlorine substituent) for 3, 4, 5 and 6 chlorine substituted compounds including more than ten kinds of isomerides. In this paper, the description of molecular structure and feature selection have been investigated. Artificial neural network (ANN) is used to relate the molecular structure and retention time of PCDD. The predicted results of 49 PCDDs retention time by ANN are listed. It was shown that as to the relative error more than 95% data was less than 10% and more than 80% data was less than 5%. PMID- 12549148 TI - [The effect of organic modifier on the capacity factor of the solute in the micellar electrokinetic capillary chromatography]. AB - In this paper the mass action law has been used to study the relationship of solute, organic modifier and micelle in the micellar electrokinetic capillary chromatography. A series of equations were given showing the relationship of capacity factor(k) of the solute, equivalent constant, concentration of solute and migration speed under the condition of using only one organic modifier. The results showed that the organic modifier can, to some extent, be dissolved in micelles. It has almost no effect on micellar actual migration speed, but, the organic modifier influences the CMC of surfactants and the distribution of solutes between two phases and results in the change of the capacity factors of the solutes. No simple equations like those in HPLC can be used to show the effect of organic modifier and surfactants on k. At the first approximation, they have the following equation, lnk approximately a + bCB + clnCB where, CB is the total concentration of organic modifier, a, b and c are constants. PMID- 12549149 TI - [Screening and analysis of biologically active components in traditional Chinese medicine by molecular biochromatography]. AB - Screening of biologically active components from Chinese medicines by molecular biochromatography has been first proposed by authors. This paper summarizes their recent progresses on screening and analysis of Chinese medicines by molecular chromatography, including the comparison of the chromatograpic pattern for a number of herbal medicines, and the same kind of medicines but different sources, method development for screening and quality control of Chinese medicines, and the study on the interactions between the biologically active compounds in Chinese medicines and protein. The prospect of molecular biochromatography on study of Chinese medicines has also been highlighted. PMID- 12549150 TI - [Intersection point rule for the retention value with mobile phase composition and boiling point of the homologues and chlorobenzenes in soil leaching column chromatography]. AB - Based on the linear retention equation of the logarithm of the capacity factor (logk') vs. the methanol volume fraction (psi) of aqueous binary mobile phase in soil leaching column chromatography, the intersection point rule for the logk' of homologues and weak polar chlorobenzenes, with psi, as well as with boiling point, has been derived due to existence of the similar interactions among solutes of the same series, stationary phase (soil) and eluent (methanol-water). These rules were testified by experimental data of homologues (n-alkylbenzenes, methylbenzenes) and weak polar chlorobenzenes. PMID- 12549151 TI - [Determination of 2,4-dinitrophenol by solid-phase microextraction coupled to GC/MS]. AB - Solid-phase microextraction coupled GC/MS was investigated for the determination of 2,4-dinitrophenol in aqueous sample. Analytes were extracted by a 85 microns polyacrylate coated fiber. The fiber was then transferred into the injector of GC. Analytes were thermally desorbed and detected by GC/MS. The influences of several parameters on SPME procedure have been studied. The optimization condition was obtained: directly sampling in the water sample with the extraction period was 30 min and the desorption time 3 min at 270 degrees C. Acidification by hydrochloric acid (pH 2) and saturated salt were necessary for the best sensitivity. The precision of SPME is better than that of liquid-liquid extraction. This method is a fast, simple, and pre-concentration-free technique, which is suitable for detecting 2,4-dinitrophenol in water sample. PMID- 12549152 TI - [Albumin-drug binding study by capillary electrophoresis. I. Quantitative applicability examination of liquid pre-column]. AB - To measure the free concentration of verapamil (a basic drug) enantiomers in the binding system of human serum albumin(HSA), a capillary electrophoretic method, liquid precolumn(LCP), was established, and the method was examined systematically. In physiological pH condition (pH 7.4, ionic strength 0.17), HSA migrates in the opposite direction of verapamil. This electrophoretic property basically supposed the probability of preventing HSA from entering the capillary whereas a positive electric field was used. Finally, the drug enantiomers were separated by the chiral selector (45 mmol/L trimethyl-beta-cyclodextrin, pH 2.5 phosphate buffer) and the free concentration of each optical isomer in the binding system was measured. Seven samples were examined and their relative standard deviations(RSD) and the relative errors (RE) of unbound drug were 2.1% 5.02% and 1.4%-5.8%, respectively. PMID- 12549153 TI - [Albumin-drug binding study by capillary electrophoresis. II. Determination of drug enantiomeric binding constants]. AB - A capillary electrophoresis (CE) method was applied to determine the binding constants of the basic racemic drug, verapamil (VER) to human serum albumin (HSA) under drug-HSA binding equilibrium (in phosphae buffer pH 7.4, ionic strength = 0.17). In coated capillary, the unbound basic drug eluted as two zonal plateau peaks due to enantiomers separated by chiral selector (45 mmol/L trimethyl-beta cyclodextrin dissolved in pH 2.5 phosphate buffer) at 15 kV, and their concentrations can be determined from the peak heights. To avoid disturbing the VER-HSA equilibrium, the pH 7.4 solution was used as the inlet vial buffer, and a plug(about 3 cm long) of this buffer was introduced to the capillary before injection of analyte. The binding constants were obtained from linear regression plots. The unbound concentration of S-VER was 1.67 times higher than that of the antipode for the solution 300 mumol/L (+/-) VER-500 mumol/L HSA, while 1.13 for 100 mumol/L (+/-) VER-100 mumol/L HSA. The study confidently provides the binding constants of VER enantiomers to HSA, which are KR = 2.7 x 10(3) (+/- 4.4 x 10(2)) and Ks = 8.5 x 10(3) (+/- 1.0 x 10(2)). PMID- 12549154 TI - [Preparation and evaluation of open tubular capillary electrochromatographic columns by sol-gel method]. AB - Open-tubular capillary electrochromatographic columns were prepared by sol-gel method. The influence of pH of gel solution on the column performance was investigated with a series of gel solution prepared by hydrochloric acid at different pH value. The results show that the pH of gel solution influences the capacity factors (k') of solutes in a complex way. The general trend is that the values of k' increase as the pH value increases except that at pH of 2.0. The gel solution of pH 2.8 was selected to prepare the column, at which the greatest capacity factors can be obtained within an acceptable reaction rate. The influence of reaction time of gel solution in column was also investigated. Out of expectation, the k' value didn't increase with increase in reaction time. The reason may be that the pore of silica gel is so small that the sample molecule can't migrate into it. The surface area is only offered by outer surface of silica gel, so the phase ratio of column prepared by this method is still not large enough. It was found that the joule heat effect can be neglected since the capillaries used were very small in inner diameter. The electroosmotic flow plays an important role in the separation as it determines the separation time. In order to accelerate the separation, high pH condition is necessary. Therefore, the stability of column under high pH is very important. The column shows a good stability under pH 9 of the mobile phase. The relative standard deviations (RSD) of capacity factor of four solutes were less than 3.7% for 100 times of operations. The separation efficiency was evaluated and the average theoretical plate numbers under optimal conditions for thiourea, phenetole, naphthalene, biphenyl, and 2,6-dimethyl naphthalene were 4.66 x 10(5), 5.24 x 10(5), 5.02 x 10(5), 4.77 x 10(5), and 3.58 x 10(5)/m, respectively. The column shows good repeatability. The RSD values for k' are less than 1% and the RSD values for efficiency are less than 8% (n = 5). In addition, fast separation was obtained with shorter column and higher voltage used, where four aromatic compounds were baseline separated in 101 s. PMID- 12549155 TI - [Effects of temperature on the migration time window of micellar electrokinetic capillary chromatography]. AB - The effects of capillary temperature, the nature and concentration of buffer on the migration time window of micellar electrokinetic capillary chromatography are investigated through three parameters including electroosmotic mobility (micro eo), the electrophoretic mobility micelle (micro ep,mc) and mobility ratio (Rm,mc). (micro eo) and (micro ep,mc) increase linearly as capillary temperature increases. Change in the viscosity of the running buffer is the main factor in the temperature effect. It is proved theoretically that the local viscosity at the capillary wall is different from the bulk viscosity. The increasing rate of micro eo with temperature is different from that of micro ep,mc. The migration time window can be expanded by lowering capillary temperature. Although the expanding is limited, this method can be very easily achieved on a commercial apparatus. A uniform formula has been derived to describe the relationships between micro eo, micro ep,mc and the concentration of buffer. For Tris and borate buffer, both micro eo and micro ep,mc decrease with increasing buffer concentration, but the decreasing rate is limited. Thus the effect on the migration time window is not significant. For phosphate buffer, the effects on micro eo and micro ep,mc are different in different concentration ranges. micro eo decreases rapidly in low concentration range but decreases slowly in high concentration range with increasing buffer concentration. micro ep,mc decreases in low concentration range but increases in high concentration range with increasing buffer concentration. The migration time window can be significantly expanded by increasing the concentration of phosphate buffer. PMID- 12549156 TI - [Chiral resolution by capillary electrophoresis]. AB - The application and development in the field of chiral resolution by capillary electrophoresis are reviewed according to recent references, including different operating models and different chiral selectors. The study on mechanism of chiral resolution is summarized as well. It appears that capillary electrophoresis is one of the most effective, fast and simple methods in chiral resolution. PMID- 12549157 TI - [Separation and detection of tryptophan metabolites in biological samples]. AB - A reversed phase HPLC method for separation and determination of the major tryptophan (TRP) metabolites in both pyrrolas pathway and TRP hydroxylase pathway, including TRP, kynurenine(KN), 3-hydroxykynurenine(3-HKN), kynurenic acid(KA), xanthurenic acid(XA), 5-hydroxytryptophan(5-HTP), 5-hydroxytryptamin(5 HT) and 5-hydroxyindoleacetic acid(5-HIAA), has been developed by sequential optimization of mobile phase based on acetate buffer and methanol. Trichloroacetic acid(TCA) was used as ion-pairing reagent to increase the retention of 3-HKN. The effects of pH and concentrations of TCA on separation were studied. Good separation can be achieved at pH 4.0-5.0 of mobile phase in less than 25 min. When TCA was not used, 3-HKN was hard to be detected in biological samples. The maximum retention of 3-HKN was obtained at pH 4.0 with mobile phase containing 50 mmol/L TCA. Combination of electrochemical (ED) and ultraviolet (UV) detection was used in which ED was responsible for detection of 3-HKN, 5-HTP, 5-HT and 5-HIAA and UV for the others. The influences of potential of ED and wavelength of UV on detection were studied. The optimization of conditions for separation and detection in different biological samples was also discussed. PMID- 12549158 TI - [Study of the distribution and migration laws of nitroglycerin in energy increasing propellant by size exclusion chromatography]. AB - It is very important to increase the energy of high-nitrogen-content single-base propellant by absorbing nitroglycerin in the case of keeping the advantage of the original propellant. In this paper, the distribution laws and migration laws in the aging of nitroglycerin have been studied by size exclusion chromatography (SEC) after high-nitrogen-content single-base propellant absorbing nitroglycerin. The results show that the distribution of nitroglycerin in high-nitrogen-content sigle-base propellant follows the Gauss distribution. In aging, this distribution changes towards the big variance Gauss distribution. The relationship between migration mass and aging time is y = k1e-k2/t, for external layer k1 = 0.1208 and k2 = 17.229, for inner hole k1 = 0.0348 and k2 = 10.97. The migration velocity constant, K = A0e-E/RT, of nitroglycerin is also given in this article, where A0 = 0.4086 and E0 = 1.6702 kJ/mol for inner hole, A0 = 31.2775 and E0 = 3.5428 kJ/mol for external layer. PMID- 12549159 TI - [Chiral drug separation by supercritical fluid chromatography]. AB - Because of the high-efficiency, fast separation, low-temperature analysis and applicable to a wide variety of detectors, supercritical fluid chromatography(SFC) is an attractive alternative for chiral drug separation. The development and application of this technique in enantiomer separation are reviewed with 48 references. The separation modes and the choices of experimental condition are discussed. PMID- 12549160 TI - [An introduction of solvent-free(SPME) technique]. AB - Solid-phase microextraction (SPME) is a solvent-free sample preparation technique in which a fused silica fiber coated with polymeric organic liquid is introduced into the sample. In this paper, theoretical aspects, operating models, factors of influencing sensitivity, applications and future development are introduced briefly with 19 references. PMID- 12549161 TI - [Application of derivative technique in the signal processing of gas chromatography]. AB - An application of derivative technique in the signal processing of gas chromatography(GC) is described. TCD's sensitivity of the derivative GC was 270 kV.L/kg, and its susceptibility was 0.3 microgram/L In comparison with the conventional GC, there was a 7-40 fold increase in sensitivity and a 24-40 fold decrease in susceptibility. The noise of the derivative GC was equal to or smaller than the conventional one. The minimum resolution for a pair of substances most difficulty to be separated in conventional GC, which was separated completely in the derivative GC, was 2.8. The linear range of n-butanol was 10(2) and the peak area correlation coefficient of the derivative GC was 0.9991. PMID- 12549162 TI - [Study on qualitative analysis method of high performance liquid chromatography by simultaneous multi-wavelength detection]. AB - In this paper, we introduce an HPLC qualitative method, that is, multiwavelength detection qualitative analysis method. According to Beer's law, for a certain compounds the ratio of peak areas at different wavelengths is a constant. Comparing the ratio of peak areas of a component at different wavelengths with that of the standard by means of simultaneous multi-wavelength detection, we established a convenient qualitative method. This method has been applied successfully to qualitative analysis of 18 aromatic amines in the prohibited azo dyes. PMID- 12549164 TI - [Determination of alpha-acid in hop by high performance liquid chromatography]. AB - A method for the determination of alpha-acid in hop by reversed-phase high performance liquid chromatgraphy is presented. The alpha-acid in hop and hop exteact was separated in a mu-Bondapak C18 cartridge by using V(CH3OH):V(H2O):V(H3PO4) = 80:20:0.25 as mobile phase. The flow rate was 1.0 mL/min and detection of alpha-acid was performed with UV detector at 314 nm. Quantification of alpha-acid was carried out by measuring peak height/peak area and comparing with external standard. The relative standard deviation of alpha acid in hop was 2.02% and in hop extract was 1.78%. The method is simple, rapid and satisfactory. PMID- 12549163 TI - [Chiral separation of thio-glycidyl ether on amylose-chiral stationary phase by high performance liquid chromatography]. AB - In the present study, we synthesized amylose-tris(phenylcarbamate), and used it as chiral stationary phase based on silica gel. This stationary phase offered a practically useful chiral separation medium. Amylose was allowed to react with an excess of phenylisocyanate in pyridine for 48 h. The obtained carbamate was dissolved in DMF and adsorbed on silica gel previously treated with 3 aminopropyltriethoxysilane. The mass ratio of the carbamate to the silica gel was 0.45/2.55. The four thio-glycidyl ethers were resolved on the amylose tris(phenylcarbamate) chiral stationary phase. Separation was carried out with a hexane/2-propanol mixture as mobile phase at a flow rate of 1.0 mL/min at room temperature. The effects of 2-propanol concentration on the retention and resolution have been investigated. PMID- 12549165 TI - [Separation of levonorgestrel, progesterone and testosterone by micellar electrokinetic capillary chromatography]. AB - Separation of levonorgestrel, progesterone and testosterone was achieved by using micellar electrokinetic capillary chromatography (MECC). A column of 53 cm (to the detector) x 50 microns i.d. uncoated fused-silica capillary (totally 68 cm in length) and an ultra-violet detector with fixed wavelength at 254 nm were used throughout all the experiments. MECC analysis was optimized by evaluating three different micelle-forming agents, the concentration of SDS and several of organic additives in a 10 mmol/L borate running buffer (pH 9.2). Complete separations were obtained with either 10%-20% acetonitrile or 20 mmol/L 2,6-dimethyl-beta cyclodextrin (DM-beta-CD) as the modifier in buffer. When the acetonitrile volume fraction was in the range of 0-15%, the migration times of the three steroids increased with the acetonitrile volume fraction, but in the range of 15%-20% acetonitrile concentrations, the situation was opposite. This behavior of the steroids was attributed to the interaction of two opposite effects, an increased mobility due to decreased partition coefficient and a decreased electroosmotic flow (EOF). Both beta-cyclodextrin (beta-CD) and gamma-cyclodextrin (gamma-CD) were found to be inadequate for a satisfactory separation. PMID- 12549166 TI - [An electrochemical cell for end-column amperometric detection in capillary electrophoresis]. AB - With end-column electrochemical detection in capillary electrophoresis(CE), precise positioning and stabilization of the working electrode are very important. In this paper, an electrochemical cell designed for end-colunm wall jet amperometric detection in capillary zone electrophoresis is reported. In this cell, a curved surface was formed because of the surface tension and used as the amplifier. With this cell, alignment of the working electrode with the capillary outlet could be achieved precisely and easily without the help of a microscope and a micropositioner. Being compared with electrochemical cell of other groups, this cell is simple and can be used more conveniently. Application of this electrochemical cell for electrophoretic separation of phenols is demonstrated. High resolution and low detection limits were obtained with this assembly. PMID- 12549167 TI - [Determination of chlorogenic acid, glycyrrhizin and glycyrrhetinic acid in yinqiaojiedupian by high performance capillary electrophoresis]. AB - A simple, rapid and reproducible capillary electrophoretic method was developed for the determination of chlorogenic acid, glycyrrhizin and glycyrrhetinic acid in Yinqiaojiedupian. The buffer solution used in this method was a mixture of 20 mmol/L NaH2PO4 and 5 mmol/L Na2B4O7(pH 7.0). The other conditions were as follows: applied voltage, 20 kV (positive to negative polarity); L = 60 cm; Leff = 55 cm; i.d. = 75 microns; lambda = 254 nm; temperature, 25 degrees C; injection time, 5 s hydrostatic. The linear calibration range was 1.568-100.4 mg/L for glycyrrhetinic acid, 8.000-128.0 mg/L for glycyrrhizin and 8.080-129.3 mg/L for chlorogenic acid, and recoveries were 98.0%-103% for glycyrrhetinic acid, 97.3% 102% for glycyrrhizin and 97.3%-100% for chlorogenic acid. The relative standard deviations (n = 3) were 0.2%-1.3% (intra-day) and 1.9%-3.0% (inter-day) for glycyrrhetinic acid, 0.5%-1.2%(intra-day) and 1.9%-2.1%(inter-day) for glycyrrhizin and 0.7%-1.3%(intra-day) and 1.6%-2.1%(inter-day) for chlorogenic acid. The contents of these three acids in Yinqiaojiedupian were successfully determined within 10 min. PMID- 12549168 TI - [Isotachophoresis analysis of hydrolytic products of DFP catalyzed by DFPase in porcine liver]. AB - DFPase can hydrolyze the P-F bond of diisopropyl phosphate fluoride (DFP) to produce diisopropyl phosphate and HF. In this article, the activity of two kinds of DFPase from porcine liver was assayed by the methods of titration and F- ion sensitive electrode. The results showed that the activity of large molecular DFPase can be determined by both methods, but the activity of small molecular DFPase can only be determined by the titration and cannot be determined by F- ion sensitive electrode. The isotachophoresis analysis of the products of DFP hydrolyzed by the large and small molecular DFPase was studied. The results showed that the small molecular enzyme could catalyze the hydrolysis of DFP, but without the production of F- ion, and that this enzyme is one kind of phosphoesterase that hydrolyzed P-OR bond of DFP. PMID- 12549169 TI - [Spherical phenyl-regenerated cellulose stationary phases used for hydrophobic chromatography to extract bramelain from crude stem bramelain]. AB - The low-pressure preparative hydrophobic chromatography packing materials have been successfully synthesized by grafting hydrophobic phenyl group to the macro pore Spherical Regenerated Cellulose Resin (diameter 0.7-1.3 mm, pore size 4.3 15.7 microns, specific surface area 88 m2/g). The optimum adsorption-desorption conditions have been got by using 10 mL of this hydrophobic resin to deal with 100 mL of the crude stem bramelain: the mobile phase pH is around 5.0, concentration of (NH4)2SO4 is 1.5 mol/L; and the pH of total eluate is 8.0. While the hydrophobic column is operated according to the following steps: 1. use 0.1 mol/L NaAc-HAc(pH 4.8) and 1.5 mol/L (NH4)2SO4 solution to soak resin for more than 30 min; 2. let 100 mL of the extracted liquor of pineapple skin, which contains the same concentration of salt as 1, pass through the hydrophobic column; 3. use 20 mL of the same solution as 1 to wash the impurities in proteins, which are not adsorbed by the hydrophobic column; 4. use 30 mL of eluant, which contains 67 mmol/L NaOH and 50 mmol/L Na2CO3-NaHCO3(pH 10.28), to elute the adsorbed bramelain, and the initial 5 mL of the eluate is discarded; 5. soak the hydrophobic column with 0.5 mol/L NaOH + 20% (volume fraction) C2H5OH for 30 min, then wash it with water, and soak it with 0.5 mol/L HCl for 30 min, at last use water to wash it to neutrality for next use. We can get enriched bramelain of 4.4 fold activity, moreover the total enzyme activity recovery is more than 120%. The Phenyl-Spherical Regenerated Cellulose Resin can be used at the pH from 1 to 13; As it has a high rigidity and big size, it can be operated under low-pressure condition; It has a larger specific surface area and a higher separation efficiency for natural bramelain; After it has been used repeatedly for more than 50 times, its efficiency does not fall. Moreover, it is cheap to be produced. PMID- 12549170 TI - [The determination of coumarin in foods by high performance liquid chromatography (HPLC)]. AB - An HPLC procedure for determination of coumarin in foods has been developed. The samples were extracted with ether. The extract was evaporated at 40 degrees C under dry N2. The residue was diluted by adding 10.0 mL MeOH-H2O(9:1) and keep standing. A 10 microL clear liquid was analysed by HPLC on a Nova Pak C18 column (5 microns, 150 mm x 3.9 mm. i.d.) with MeOH/10 mmol/L-phosphate buffer of pH 5.4(45:55) as mobile phase (1.0 mL/min) and UV detection at 275 nm. The calibration curve was linear in the range from 2 to 10 mg/L and the detection limit was 0.015 mg/L. Recoveries of the method were from 97.6% to 100.8%. The CV (n = 6) was 1.3%. The method is simple, rapid and accurate. PMID- 12549171 TI - [Determination of sulfacetamide sodium and sulfanilamide in shao tang ling ointment by high performance liquid chromatography]. AB - An HPLC method for the determination of sulfacetamide sodium and sulfanilamide in Shao Tang Ling ointment was developed. The chromatographic conditions were: column, Spherisorb C18, mobile phase, methanol-0.25% (V/V)-acetic acid solution (7:93); detector UV-257 nm. The average recoveries were 98.9% (n = 5), RSD = 0.64% for sulfacetamide sodium and 99.3% (n = 5), RSD = 1.58% for sulfanilamide. The analytical method is simple, quick, accurate and reproducible. PMID- 12549172 TI - [Application of high performance liquid chromatography to the study on the production of maltooligosaccharide by synergistic action of two-enzymes]. AB - This paper describes the application of high performance liquid chromatograpy to the study on the production of maltooligosaccharide by synergistic action of two enzymes. Seven sugars in products were separated on a column of Nucleosil C18 with water as the mobile phase and determined refractometrically. Meanwhile, for the preparation of maltooligosaccharide, the effect of supplementing rate of the two enzymes on the content of maltooligosaccharide in products is examined in this paper. The optimun technological conditions were obtained by using the orthogonal design and the amount of the two enzymes added was determined. Thus more than 70% of M3-M6 maltooligosaccharide in product was obtained. PMID- 12549173 TI - [Determination of active components in caoyiguang wettable powder by reversed phase high performance liquid chromatography]. AB - The Caoyiguang wettable powder is composed of two active components, synergists and other additives. The active components, bensulfuron methyl (Londax) and metolachlor (Dual), in 15.5% Caoyiguang wettable powder were determined by reversed phase high performance liquid chromatographic method simultaneously. The ZORBAX SB-C18 column (4.6 mm i.d. x 250 mm, 5 microns) and acidic mixture of methanol and water as mobile phase were used. The results showed that the recoveries were 98.1%-102.6%, and the coefficients of variation were 0.4%-0.8%. The method has been used to determine samples with satisfactory results. PMID- 12549174 TI - [Identification of corrected retention indices of ten phenols in reversed-phase high performance liquid chromatography]. AB - Corrected retention indices in RP-HPLC of ten phenols based on 2-keto-alkanes were examined on three ODS-silica columns. The RI values for phenols showed large variations. However, when the correction was applied, the variations diminished significantly. It is clear that the correction procedure improves the variation of phenols' RI values obtained from different ODS columns. The procedure is recommended as a method of standardization of HPLC data. PMID- 12549175 TI - [The enantiomeric separation of aromatic alcohol amino drugs by thin-layer chromatography]. AB - Two chiral aromatic alcohol amino drugs, Labarol and Bataroc, were resolved by thinlayer chromatography (TLC) on the silica gel GF254 plates (2.5 cm x 10 cm), by using the ammonium-D-10-camphorsulfonate (CSA) as chiral ion-pair interaction agent which was added to the mobile phase in the ammonium form. All developments were carried out at lower temperature (2-4 degrees C, in a refrigerator) in small glass jars of 250 mL volume. These two drugs were not resolved at room temperature (15-30 degrees C). Analytical reagent grade methanol and dichloromethane can be directly used as mobile phase without further drying. The chiral separation occured over a range from 40% to 70% (optimum 67%) dichloromethane in the mobile phase volume ratio and 55% to 80% (optimum 60%) dichloromethane in the mobile phase volume ratio. These separation conditions were easily obtained. This method is relatively inexpensive and attractive. PMID- 12549176 TI - [Quantitative analysis of butachlor, oxadiazon and simetryn by gas chromatography]. AB - The quantitative analysis of the ingredients in 26% B-O-S (butachlor, oxadiazon and simetryn) emulsion by gas chromatographic method was carried out with a 5% SE 30 on Chromosorb AW DMCS, 2 m x 3 mm i.d., glass column at column temperature of 210 degrees C and detector temperature of 230 degrees C. The internal standard is di-n-butyl sebacate. The retentions of simetryn, internal standard, butachlor and oxadiazon were 6.5, 8.3, 9.9 and 11.9 min respectively. This method has a recovery of 98.62%-100.77% and the coefficients of variation of this analysis of butachlor, oxadiazon and simetryn were 0.46%, 0.32% and 0.57% respectively. All coefficients of linear correlation were higher than 0.999. PMID- 12549177 TI - [Gas chromatographic method for estimating chemical stability of nitroquanidine propellant]. AB - This paper describes a GC analytical method for nitroquanidine propellant instead of the tranditional methylviolet method. Operating conditions were as follows: TDX-01 packed column, 150-200 mesh, 800 mm x 3 mm i.d., at 150-170 degrees C with TCD. Released gases, CO2 and N2O contents in a sealed vessel packed with nitroquanidine were calculated according to the normalization of corrected peak areas of air, CO2 and N2O. Thus, the change in the chemical stability of nitroquanidine during the period of storage and thermal aging could be observed. PMID- 12549178 TI - [Gas chromatography/mass spectrometry analysis for determination of methyl alpha (6-methoxyl-2-naphthyl) propionate from carbonylation]. AB - alpha-(6-Methoxyl-2-naphthyl) ethanol can be carbonylated to form methyl-alpha-(6 methoxyl-2-naphthyl)propionate catalyzed by a catalyst in the pressnce of methanol. High efficiency capillary gas chromatography and GC/MS technique were used for determining products which were obtained from the carbonylation. An OV 101 fused silica capillary column (25 m x 0.2 mm i.d.) and a flame ionization detector (FID) were employed in the GC analysis. The column temperature was kept at 250 degrees C. Mass spectra were obtained by electron impact at 70 eV. The structures of four compounds were identified. The major product is methyl-alpha (6-methoxy-2-naphthyl) propionate, and the two by-products are 2-vinyl-6 methoxynaphthalene and 1-methoxy-(6'-methoxy-2'-naphthyl)ethanol. PMID- 12549179 TI - [Determination of 5-hydroxytryptamine(5-HT) secretion from activated platelets by high performance liquid chromatography/electrochemical detector]. AB - A method of determination of 5-HT secretion from activated platelets by HPLC/ECD was established. Spherisorb C18 column of 4.6 mm i.d. x 100 mm was used in this experiment. The CV was 4.65%, The linear range was 0.002 5-1 mg/L. This method has the advantages of high speed, high sensitivity; specificity, of requiring a minimum of preparation of samples and of relatively low cost for instrumentation and reagents. It could reflect the activated situation of platelet induced by weak activator and be used in other biological sample. PMID- 12549180 TI - [What are the opportunities and challenges facing us?]. PMID- 12549181 TI - [Some biological characteristics of genetically engineered insecticidal Pseudomonas fluorescens]. AB - Plasmid stability, Antifungal activity, plant-colonizing ability, UV resistance and inseticidal activity in field were analysed for the engineered Pseudomonas fluorescens (Pf) strain IPP202. The results indicated that the recombinant plasmid in IPP202 was very stable after successive diluting culturing and after continuous culturing, There was no significant change in the properties beneficial to plants, such as antifungal activity and plant-colonizing ability as compared with the original strain P303. IPP202 was much more resistant to UV than Bt strain HD73. The control effect in field against cotton boll worm in field was close to that of a locally used Bt-chemical mixture in normal applied concentration. All the data indicated that the engineered Pf strain was a one with prosperous future after further study. PMID- 12549182 TI - [Subcloning and sequencing of DNA fragment related to salt tolerance in Sinorhizobium meliloti 042B]. AB - A 4 kb ClaI DNA fragment related to salt tolerance from S. meliloti 042B was digested by HindIII down 2.4 kb fragment, and a 1.6 kb ClaII-HindIII fragment was retained on plasmid pML122. Then, the 2.4 kb DNA fragment was ligated with plasmid pBBR1-MCS2, and the recombinant plasmid was transformed to E. coli DH5 alpha, and transformant GS2 was obtained. Three-parental mating experiments were carried out with transformant GS2 as donor, salt sensitive strains GZ17 as recipient and pRK2013 as helper plasmid, then the transconjugant GG2 was selected on FY plates containing kanamycin and 0.4 mol/L NaCl. The remaining DNA fragment was self ligated with pML122 and then transformed into E. coli S17-1 and transformat GS0 was obtained. Two-parental mating experiment was carried out with transformant GS0 as donor and salt sensitive strain GZ17 as recipient, but no transconjugant was obtained on the FY plates. Then, the 2.4 kb HindIII DNA fragment was ligated into sequencing vector pGEM-7Zf(+) for sequencing. The result of sequencing and analysis showed that the 2.4 kb DNA fragment contained three ORFs. According to the result of sequencing, further subcloning was conducted and 1.9 kb HindIII-Sac II DNA fragment related to salt tolerance was obtained. PMID- 12549183 TI - [Construction of recombinant E. coli with high glutathione biosynthetic activity and the biosynthetic process]. AB - A recombinant strain E. coli II-1, which exhibited high glutathione (GSH) biosynthetic activity and high stability, was constructed by transforming plasmid pGH501 which contains gene gsh I and gsh II into a wild type strain E. coli II. 4 g/L GSH accumulated extracellularly by using toluene-treated cell. In GSH biosynthetic system, GSH production was improved by increasing the concentration of L-glutamate, while inhibited by L-cysteine if it's concentration was beyond 20 mmol/L. In GSH biosynthetic reaction, the apparent little consumption of L glutamate and glycine was concluded experimentally to be that toluene-treated E. coli II-1 cells still contained high concentration of L-glutamate and glycine. According to the change of energy cofactor in the GSH biosynthetic process, a possible GSH biosynthetic mechanism controlled by E. coli II-1, was proposed: the energy donator of reaction catalyzed by glutathione synthetase (GSH-II) was ADP but not ATP, the reaction was rate-limited step within the whole GSH biosynthetic process, high concentration of ADP might inhibit the activity of GSH-II. Further degradation of GSH was prevented by the addition of 100 mmol/L L-serine and potassium borate mixture. In such case, 23.0 mmol/L (about 7.1 g/L) GSH accumulated at 3 h. PMID- 12549184 TI - [Sequence analysis of RAN4 of a severe isolate of rice stripe virus in China]. AB - RNA4 segment of a severe isolate of Rice stripe virus, isolated from Liaoning province and designated as PJ isolate, was amplified, cloned and sequenced. RNA4 of PJ isolate had 2157 nucleotide in length. When compared with RNA4 of T and M isolates of Japan and CX isolate of Yunnan province of China that had been previously reported, we found that these four isolates could be divided into two groups. PJ, T and M isolates shared 97.0% and 97.0%-97.5% identities in vORF4 and vcOFR4 at the nucleotide level, respectively and formed one group. The sequences in 5' and 3' terminal non-encoding region were completely identical among these three isolates. In this group, PJ isolate was more closely related to T isolate than to M isolate. The length of intergenic region(IR4) of PJ isolate was as same as that of T isolate, and had 93.0% sequence identity. However, PJ IR4 had an insertion of 19 bp in length compared with isolate M and had only 85.0% sequence identity. CX isolate belonged to another group, which shared only 94.0% and 92.5% 93.5% sequences identities in vORF4 and vcORF4 at the nucleotide level, respectively, even though there were not significant difference between these two group at the amino acid level. There was an insertion of 84 bp in length in the IR4 of CX isolate compared with PJ isolate and the sequence identity between two group reached to 72.0%-75.0%. Even though no base variation occurred in 5' terminal non-coding region, there was two bases substitution in 3' terminal non coding region. These results showed that the isolates were grouped according to their geographical location. Additionally, highly consensus in 5' and 3' non encoding region suggested that these regions played an very important role in transcription and replication of viral genome. Finally, the molecular epidemiology and gene functions of Rice stripe virus were discussed in this paper. PMID- 12549185 TI - [Cloning and sequencing of Chinese squash leaf curl virus]. AB - Sequence analysis of virus isolation DNA of Squash leaf curl disease shows that Squash Leaf Curl Virus (SLCH-CHI) contains 2741nt, which encode six Open Reading Frames (ORFs). Two of them are on virus sense strand: AV1 (256aa) and AV2 (140aa). AV1 encode coat protein. The other of ORFs are on complementary sense strand: AC1 (replicase gene, 243aa), AC2 (transactivtor, 134aa), AC3 (136aa) and AC4 (172aa). The virus belongs to one number of begomoviruses from old world. PMID- 12549186 TI - [Expression of human interleukin-12 in Baculovirus expression system]. AB - Human Interleukin-12 (hIL-12) is a key immunoregulatory cytokine and the only heterodimeric cytokine found by far. It is a disulfide-linked heterodimer composed of a 35 kD light chain (P35) and a 40 kD heavy chain (P40). Two recombinant transfer vector plasmids pAcAB3-p35 (including hIL-12 p35 gene) and pAcAB3-p40 (including hIL-12 p40 gene) were constructed and co-transfected the insect cells Sf9 with AcNPV BaculoGold Linearized Baculovirus DNA. The obtained recombinant viruses, AcNPV-OCC(-)-hiL-12 (p35) and AcNPV-OCC(-)-hIL-12 (p40), were used to infect the Sf9 cells separately, and the supernatants of culture and cell lysate were examined by using SDS-PAGE and Western blot. Results showed that both hIL-12 p35 and p40 gene were successfully expressed in insect cells and secreted. The expression productions had immunogenicity. PMID- 12549187 TI - [Construction of rice dwarf virus genome database]. AB - Secondary database construction is an important subject in the field of bioinformatics. As the full genomic sequences of some organisms are being completed and followed by structural and functional studies, construction of secondary database becomes essential on the agenda. The rice dwarf virus (RDV) is a pathogen infecting rice in China, Japan and the Southeastern Asia region and leading to considerable economic loss. Based on the data generated from recent genomic research and earlier biochemical studies scattered in various primary databases and scientific journals, we have constructed a compact, user-friendly and non-redundant job-oriented secondary database. This work will provide compiled useful information for plant molecular biologists as well as in achieving preliminary experiences in secondary database construction. PMID- 12549188 TI - [Determination of chromosome of Gluconobacter oxydans SCB329]. AB - After the pure culture of Gluconobacter oxydans SCB329 was researched, its growth curve was measured and its logarithmic phase was determined as between 4-24 h. After the microorganisms were havested in its logarithmic phase, The intact chromosome was prepared by agarose-embedded method. Then the genome of SCB329 was analyzed by Pulsed-field Gel Electrophoresis. The result indicated that there are one chromosome and one great plasmid. The length of intact chromosome of SCB329 has been estimated to be approximately between 2.2 Mb and 3.5 Mb. PMID- 12549189 TI - [Cloning of trehalose-6-phosphate synthase gene from S. cerevisiae and its plant expression vector construction]. AB - Total RNA, mRNA were isolated from baker's yeast S. cerevisiae, the cDNA was prepared with AMV Reverse Transcriptase from the total mRNA. The gene of trehalose-6-phosphate synthase was cloned from the cDNA with PCR amplification. The gene was sequenced and the results showed that the tpsl gene contains 1507 nucleotides and 99.6% identity with S. cerevisiae. The tps1 gene was constructed on the plant expression vector pBin438. PMID- 12549190 TI - [Structure and function of sanL--a gene involved in nikkomycin biosynthesis of Streptomyces ansochromogenes]. AB - The chromosome walking strategy has been used in this experiment and a 10 kb DNA fragment was cloned from the cosmid library of Streptomyces ansochromogenes using a partial DNA fragment involved in the nikkomycin biosynthesis as a probe. The nucleotide sequence showed that the DNA fragment contains two complete open reading frames (ORFs), they are designated as sanK and sanL respectively. The sanL is located on the upstream of sanK, it has 1281 nucleotides with ATG at 345 position as start codon and TGA at 623 position as stop codon. The deduced product is L-lysine 2-aminotransferase using Blastx program. The experiment of gene disruption indicated that this gene is closely related to nikkomycin biosynthesis of Streptomyces ansochromogenes. PMID- 12549191 TI - [Construction of plasmid gene bank of V. cholerae O139 and detection of O-antigen genes]. AB - Because O-antigen biosynthesis genes are a tandem gene cluster. Gnomic fragments of 4-20 kilobases (kb) were obtained by digesting genomic DNA of V. cholerae O139 with restriction endonuclease EcoRI, then plasmid gene bank was constructed. Recombinant colony, E. coli DH5 alpha (pMG320), expressing O-antigen of V. cholerae O139 was detected from the bank by immunological agglutinative reaction. The futher analysis showed O-antigen expressed by recombinant colony had both immunogenicity and reactogenicity, and the size of O-antigen biosynthesis genes was about 4.6 kb. PMID- 12549192 TI - [Determination of DNA-DNA homology among Pseudomonas cocovenenans subsp. farinofermentans with microdilution plate hybridization method]. AB - The DNA-DNA homologies among Pseudomonas cocovenenans subsp. farinofermentans and 11 species in genus Burkholderia were determined with microdilution plate hybridization method. The results showed: the homologies among P. cocovenenans subsp. farinofermentans, B. gladioli and B. cocovenenans were all over 75%. It was advised that these three species were synonym, and should be renamed as Burkholderia gladioli. PMID- 12549193 TI - [Cloning and DNA sequencing of Bacillus pumilus endo-1,4-beta-glucanase gene]. AB - Bacillus pumilus S-27 screened from marine environment produces an extracellular endo-1,4-beta-glucanase. The enzyme has its optimal activity at pH 5.0-7.0 and 55 degrees C and still retained 79.0% of it at pH 9.0. The gene encoded this endo 1,4-beta-glucanase was cloned and sequenced. The structural gene contained an open reading frame of 1980 bp, corresponding to 660 amino acids, the amino acid sequence of this enzyme is very close to that of an EG of C. cellulovorans and an EG of Bacillus sp. KSM-522, all belong to the cellulase family E2. PMID- 12549194 TI - [Purification and properties of chitinase from Enterobacter aerogenes]. AB - A bacterium producing chitinase was isolated from the dead body of Gymephorap ruoergensis. A chitinase was isolated from the culture of E. aerogenes and purified by means of ammonium sulfate precipitation, DEAE-cellulose column chromatography, and Sephadex G-100 column gel filtration. The purified chitinase showed homogeneity on the native polyacrylamide gel electrophoresis. Its molecular weight was estimated to be about 42.5 kD by SDS-PAGE. The optimum pH and temperature for hydrolysis of chitin were 6.0 and 55 degrees C respectively. Michaelis constant was 2.88 mg/mL. Different metal ions showed different effects on the chitinase activity, The chitinase activity was enhanced by Zn2+, Ba2+, Ca2+, Mn2+ and was strongly inhibited by Hg2+, Co2+, Mg2+. PMID- 12549195 TI - [The effect of culture conditions on the fatty acid composition in the mycelium of the Cephalosporium sp]. AB - The optimal condition of Cephalosporium sp. to culture mycelia and accumulate polyunsaturated fatty acid (PUFA) was researched. The optimal culture condition to get mycelium productivity was: maltose 60 g/L, KNO3 3 g/L, initial pH 6.0, 100 mL medium in 500 mL flask, seeding 25% (v/v), 25 degrees C culture it for 10 days. The optimal condition to accumulation PUFA proportion to total fatty acid was: glucose 10-20 g/L, NH4Cl or (NH4)2SO4 3 g/L, initial pH 4.0, 100 mL medium in 500 mL falsk, seeding 10-20% (v/v) and lighted it when it was cultured. It was suggested that two step could be used in the producing progress. A proposal was put forward that the oileic acid transformed to linoleic acid was the key step to produce PUFA. This proposal gave a base to research the optimal culture condition and enzyme regulation. PMID- 12549196 TI - [Classification for two isolates of Microbispora producing immunosuppressants]. AB - Two strains named SIPI-226 and SIPI-207 producing immunosuppressants were isolated from soil samples collected in park of Wuxi city in China. Identification was done by morphological observation, chemical analysis and nucleic acid ribotyping and 16S rRNA sequence. Two strains contain meso diaminopimelic acid in cell walls, Type PIV phospholipids, a lack of mycolic acids, MK-9(H0, H2, H4) menaquinones and G + C content of 68.3% to 69.4%, and two strains have 97.2% to 98.6% homology in 16S rRNA gene sequences, and the DNA homology by Ribotyping are 53% to 70%, but differ from seven other mesophilic Microbispora rosea strains. It could be concluded that these two strains represented two new subspecies in the genus of Microbispora rosea. We designated the strains as Microbispora rosea subsp. wuxiensis and Microbispora rosea subsp. yuantouzhuensis. The type strains of M. rosea subsp. wuxiensis and M. rosea subsp. yuantouzhuensis are SIPI-226 and SIPI-207, respectively. PMID- 12549197 TI - [The construction and application of promoter-signal peptide probe vector of lactic acid bacteria]. AB - A promoter-signal peptide probe vector of lactic acid bacteria, designated pZLB based on pMG36e has been constructed. The vector contained a reporterd gene encoding for beta-lactamase, which lacked a promoter and a functional signal sequence. Sau3A restriction fragments (80-400 bp) from L. lactis ATCC 7962 total DNA were cloned into BamHI site of pZLB. A lot of different size framents were selected. These fragments promoted transcription and secretion of beta-lactamase in L. lactis. Sequence analysis revealed that inserted fragment in pZLBe7 contained typical promoter, SD sequence and atypical signal sequence. PMID- 12549199 TI - [Specific amplification of the coding sequences of BBTV III, IV, I and their application in BBTV detection]. AB - Banana bunchy top virus disease (BBTD) is a disastrous disease in bananas, and it is spreading in the world (including China) by the banana bunchy top virus(BBTV). At present, virus-free plantlets are used to prevent BBTD in banana production, therefore, it is very important to establish a method to detect BBTV quickly, sensitively and specifically. ELISA is now popularly used to detect BBTV. The sensitivity of this method is not high enough, and needs specific antiserum, otherwise, pseudo-positive results often occur. According to DNA coding sequences of component III, IV and I of BBTV isolates from Zhangzhou, China, three pairs of primers are designed to establish a PCR method to specifically amplify parts of coding sequences of the BBTV coat protein, movement protein and replicase association. This method is also applicable to detect BBTV of bananas or cultured banana seedlings in other regions. PMID- 12549198 TI - [Expression of alpha-acetolactate decarboxylase gene from Bacillus subtilis in Brewer's yeast]. AB - Yeast YIp-type expression recombinant plasmid(pCMA2-1) was constructed. The expression of alpha-acetolactate decarboxylase gene from Bacillus subtilis was controled by CUP1 promoter and its own terminator. The recombinant plasmid pCMA2 1 was introduced into the brewer's yeast PJ3-5. Transformants were selected using copper resistance as selected marker. The results of activity assay showed that PJ3-5 didn't produce alpha-acetolactate decarboxylase(ALDC), where as the activity of alpha-ALDC in transformants were induced by copper sulfate. The laboratory scale fermentation test confirmed that the total diacetyl concentration was reduced effectively by alpha-ALDC in transformant. PMID- 12549200 TI - [Analysis on 16S rDNA sequence of rhizobia isolated from Kummerowla sp]. AB - Based on the previous studies on numerical taxonomy, SDS-PAGE of whole-cell protein and DNA hybridization, the rhizobial strains isolated from Kummerowia sp. in semi-arid area of North-west constituted a new subgroup, the 16S rDNA sequence of representative strain SH714 were tested. The unrooted phylogenetic tree was produced. In this tree, the strain SH714 with Sinorhizobium xinjiangensis, S. fredii, S. meliloti, S. medicae, S. saheli and S. teranga constituted a branch of Sinorhizobium. Within this branch, the similarity valuse of 16S rDNA sequence between strain SH714 and S. xinjiangesis, S. fredii, S. meliloti, S. medicae, S. saheli and S. teranga were 97.4%, 97.5%, 96.8%, 96.7%, 97.2% and 95.6% respectively, the values were more than 95%, this indicated that these known species should belong to the same genus. The values of DNA homology between type strains of these species were less than 70%. Thus, the strain SH714 represented a new rhizobial species, and there were some diversity between SH714 and known rhizobial species in phenotypic feature and composition of protein. PMID- 12549201 TI - [Study on the cytochrome P450 activity in alkane converting process of Candida tropicalis]. AB - A method of reduced CO-difference spectrum was established to study the cytochrome P450 activity of the whole cell of Candida tropicalis during the alkane converting process. Using this method, the cytochrome P450 activities of the whole cells that were cultured in the different concentrations of alkane were studied. The results showed that the 5% alkane could induce the cytochrome P450 activity obviously but not inhibit the growth of cells, so it was determined preliminarily that the alkane concentration of the seed medium was 5%. The cytochrome P450 activities of dicarboxylic acid (DCA) fermentation processing were further studied. During the exponential phase of growth, the cytochrome P450 activity increased smoothly. However, during the phase of production of dicarboxylic acid, the cytochrome P450 activities increased rapidly after a sort decrease. The results still showed that the rate of production of dicarboxylic acid increased with the cytochrome P450 activity. PMID- 12549202 TI - [The advance on taxonomy of Streptomyces]. PMID- 12549203 TI - [Comparison observation on the mature alveolar of Echinococcus sibiricensis and Echinococcus multilocularis in the experimentally infected white mice]. AB - The alveolar echinococcus is one of the most dangerous worm parasites in man. Rausch and Schiller reported a new species, Echinococcus sibiricensis n. sp. from arctic fox, Alpex logopus, on St. Lawrence Island of Alaska, USA. According to the view of Vogel, the sibiricensis form is only a geographical race or subspecies of Europe Echinococcus multilocularis. So far, the two names, Echinococcus multiocularis multilocularis and Echinococcus multilocularis sibiricensis, existed in many references and text books. We have found the adults of Echinococcus sibiricensis and Echinococcus multilocularis from sand foxes, Vulpes corsac and their larval stages (alveolar echinococcus) from field voles, Microtus brandti in the Hulunbeier Pasture of Inner Mongolia, northeastern China in 1985 and 1998-1999. Two types of metacestodes with quite different styles of early development of E. sibiricensis and E. multilocularis were found from field voles and laboratory experimental white mice. As one characteristic of alveolar E. multilocularis, the capsules are produced by the exogenous budding of germinal cell layer together with cyst wall. The protoscoleces grow from germinal cells on germinal cell layer. The peduncles of early protoscoleces attached to the germinal cell layer on the inner surface of capsule wall(Plate I, Figs. 1-2). Some protoscoleces in reticular structure were linked with the inner surface of capsule wall (Plate I, Fig. 3) in livers of mice in 9.5th month postinfection. In 14th month old alveolar multilocularis, large number of mature protoscoleces in reticular structure were still linked to the inner surface of capsule wall (Plate I, Figs. 4-8). The cavities of some capsules were filled with protoscoleces in meshes of reticular structure which were also linked around with the inner surface of capsule wall (Plate I, Fig. 9). The superficial surface of livers of positive field voles and experimental mice never showed any hyperemic phenomenon. The superficial surfaces of livers and lungs of positive field voles and experimental mice infected with alveolar E. sibiricensis were highly hyperemic. The metacestodes of E. sibiricensis composed of mother cyst, undifferentiated embryonic cysts and small brood capsules. Cavities of all cysts were fully filled with germinal cell masses. Host reaction appeared to be very strong, all cysts were surrounded by thick connective tissue and dense leukocytes (Plate II, Fig. 10). All alveolar vesicles were found located in lungs tissue of experimental mice. Large germinal cell masses metastasized out from undifferentiated embryonic cysts into host lung tissue, where germinal cell masses developed into accumulation of early protoscoleces (Plate II, Figs. 11-12). Early protoscoleces of alveolar E. sibiricensis were seen earliest in mice lung tissues on 101-104th days after infection. Many small capsules in different sizes and different shapes containing mature protoscoleces and reticular structure (Plate II, Figs. 13-15) were found in lungs of mice in 9th month after infection. Only in one experimental mouse infected with alveolar E. sibiricensis in 8.5th month postinfection, both its lung and liver existed alveolar cysts; the capsules in liver were surrounded by very thick connective tissue of the host, and there were some protoscoleces in their cavities (Plate II, Figs. 16-18). PMID- 12549204 TI - [Differentiation of human hepatocarcinoma SMMC-7721 cells induced by HMBA]. AB - In this paper, the effects of HMBA on the differentiation of human hepatocarcinoma cell line SMMC-7721 were investigated. After treated with 5 mmol/L HMBA, the proliferation of SMMC-7721 cells was inhibited remarkably, the cell growth inhibitory rate amounted to 64.14%, the cell mitotic index was declined by 53.88%. Light microscopy and transmission electron microscopy showed that the morphology and ultrastructure of the cells treated with HMBA undergone restorational alteration. Cytochemistry and immunocytochemistry assay revealed that the activities of gamma-GT declined and the levels of AFP and PCNA downregulated while the activity of TAT increased significantly after HMBA treatment. In the meantime, flow cytometry analysis showed that HMBA could arrest the cells in G0/G1 phase. The results showed that HMBA could effectively inhibit the proliferation, reverse the malignant morphology and ultrastructure, alter the levels of enzymes and antigens, arrest the cells in G0/G1, and induce the differentiation of human hepatocarcinoma SMMC-7721 cells in vitro. PMID- 12549205 TI - [The location of new porcine microsatellites by means of PRINS and somatic cell hybrid panel]. AB - In this study, two new microsatellites HAU02 and HAU06, which are screened and cloned. HUA02 and HAU06 are repectively mapped in lq23-27 and 6q11-21 by means of PRINS (primed in situ labeling) and somatic cell hybrid (SCH) panel techniques, we discussed their advantage and disadvantage in gene mapping. These two means provide us more techniques in gene mapping, and the locations of these new microsatellites accumulate more resource for Porcine Physical Map. PMID- 12549206 TI - [The study of a HPRT locus-specific transgenic method based on FLP recombinase mediated cassette exchange]. AB - Exchange plasmid pF-HPRT-F3 and Flp expression plasmid pCMV-Flp were constructed and then introduced using electroporation system into F18 ES cell line, which have an exchange cassette F-Neo-F3 at HPRT locus. After HAT selection, HAT resistant clones were obtained. Then G418 sensitivity test and Southern blotting were carried out to screen RMCE recombinants. The results indicated that RMCE had taken place in three of 12 HAT resistant clones. The frequency is 25%. The result demonstrates that it is realizable to introduce transgene to HPRT locus by using Flp recombinase mediated cassette exchange reaction. PMID- 12549208 TI - [The presence of the lymphatic stomata in ovarian bursa of guinea pig and the ultrastructure of ovarian bursa]. AB - In order to explore its structure and speciality of species, the ovary bursa of guinea pig was studied by using dissecting microscopy and scanning electron microscopy. In this paper, the lymphatic stomata on both internal and external layer of the ovary bursa was frist reported in guinea pig. The results suggested that the stomata in bursa not only provided a pathway to connect the bursal cavity with the lymphatic vessels of bursa and the peritoneal cavity, but also may be involved in the reproduction and local immunity of the ovary. The stomata may play an important role in physiological function of the ovary. At the same time, the structural differences were identified in ovary bursa between guinea pig and golden hamster. PMID- 12549207 TI - [Immunocytochemical localization of fertilin beta on human sperm]. AB - Fertilin is a kind of sperm plasma membrane protein that mimics snake venom protein. It belongs to the ADAMs family of surface proteins that contain a disintegrin and a metalloprotease domain. Fertilin functions in the sperm-egg binding process by connecting the sperm to the egg plasma membrane via a binding site in the disintegrin domain of fertilin beta (HF93). Its localization on the sperm is in the change. In this study, the monoclonal antibody against human fertilin beta was prepared and used to analyze the localization of fertilin beta on capacitated and acrosome-reacted sperm by immunofluorescence and immunoelectron microscopy techniques. The results were as follows: (1) fertilin beta became restricted to the anterior head during the course of capacitation. (2) During the course of acrosome reaction, the expression and localization of fertilin beta changed immensely on the anterior head and restricted to the lateral of posterior head at last. The restrictions of fertilin beta to the anterior head of capacitated sperm of human beings indicated that fertilin beta may be involved in the binding the sperm to the epithelial cells of the oviduct; the restrictions of fertilin beta to the posterior head domain of acrosome reacted sperm implied its function in sperm-egg binding and fusion. PMID- 12549209 TI - [A comparative study on cytochemistry between spring and fall peanut seeds]. AB - A comparative study was conducted on cytochemistry of spring and fall crop seeds in peanut cultivars Quanhua No. 10 and Shanyou 71 respectively. Lipids, protein, and polysaccharides in cells of axis and coteledon were simultaneously shown in the Epon812 buried section by means of cytochemistry, and their morphology, quantity and distribution were compared. Embryo cells of spring crop seed develop fully with big cell more vivid contrasting texture and more regularly disposed organelle, but the counterpart cells in fall crop seeds were not as much mature and their organelle arrangement appeared somewhat irregular. In cotyledon storage cells, there were also some difference between spring crop seed and fall crop seed. Cells of spring crop seeds were full of reserves, with more lipid and protein bodies that were closely ranged and extruded with each other. However, the cell structure in fall crop seeds was more loosely arranged, vacuoles had not been filled with protein, but starch grains accumulated more. Therefore, it was shown clearly that spring crop seeds have some advantages over fall crop seeds on production application. Moreover, some cytochemical techniques for demonstration of lipid, polysaccharide and protein in thick resin section and the stain protection were discussed in the paper. PMID- 12549210 TI - [The ultrastructural study of synergids before and after fertilization in watermelon embryo sacs]. AB - The ultrastructure of synergids of watermelon (Citrullus Lanatus L.) was investigated using transmission electron microscopy at following stages of embryo sacs: 1. Unpollination, on the first flowering day. 2. Unpollination, on 2nd day after anthesis (DAA). 3. Fertilization, on DAA 2. The synergids with distinct filiform apparatus at the micropylar end have abundant organelle, such as mitochondria, endoplasmic reticulum, and plastids in cytoplasm, which indicate that they are active on the first flowering day. No wall is present at the chalazal part of synergid, and there are some flocculent materials and vesicles in the spaces of cytoplasma membranes among synergid, egg cell and central cell in embryo sacs at the first and the second stages. On DAA 2, in unpollinated embryo sacs, the central large vacuole of synergid is divided into several smaller ones and the starch grains decrease in cytoplasm. There is no newly synthesized wall at the chalazal end of persistent synergid in fertilized embryo sacs. The contents of degenerated synergid, in the form of electron dense granules, are located in the wide space among central cell, zygote and persistent synergid, and some of them migrate into central cell through cytoplasma membrane. Therefore, it is deduced that the contents of synergid might serve as a nutrient supplement to the development of endosperm, but not embryo. PMID- 12549211 TI - [Relation between plasma membrane redox activity and extracellular carbonic anhydrase activity in marine phytoplankton]. AB - Exofacial ferricyanide reduction at the plasma membrane of intact cells, and the link between plasma membrane redox activity, inorganic carbon status of the cells and extracellular carbonic anhydrase (CAext) activity were assayed using 10 marine phytoplankton species. In species Chaetocceros compressus, Cocolithus pelagicus and Gephyrocapsa ocetanica with no extracellular CA activity under carbon-limited or carbon-replete conditions, barely detectable ferricyanide reduction was observed. Species Skeletonema costatum, Melosira sp., Thalassiosira rotula, Thalassiosira weisflogi and Pleurochrysis carterae in which extracellular CA activity was only detected under carbon-limited conditions showed high rates of exofacial ferricyanide reduction. Western blotting and immunolocalization showed the presence of enzyme protein under carbon-limited and replete conditions at the cell surface, even though the CA activity could only detected when inorganic carbon was limiting, which suggests that the development of extracellular CA in response to carbon limitation is an activation of a preexisting protein rather than de novo synthesis. The results suggest that inorganic carbon limitation in the light increases plasma membrane redox activity and promotes proton extrusion, which result in the protonation and activation of the extracellular CA. PMID- 12549212 TI - [Cloning and characterization of syap1, a down regulated gene in human hepatocellular carcinoma]. AB - Using a different fragment DE6 obtained from DD-PCR as probe, a full-length cDNA has been cloned from human liver cDNA library and named as syap1 (Synapse associated protein 1) because its deduced amino acid sequence is homologous to SAP47 of D. melanogaster. Northern blot analysis reveals that two transcripts of syap1 mRNA are expressed in cultured human liver L-02 cells. Moreover, the syap1 mRNA was also detected in most of adult human tissues by RT-PCR analysis. Down regulated expression of syap1 mRNA was confirmed by semi-quantitative RT-PCR assay in 5 of 10 (50%) cases of human hepatocellular carcinoma (HCC). PMID- 12549213 TI - [Ultrastructural localization of ATPase activity in fertile and sterile anther of rice (Oryza sativa L. cv. Marxie)]. AB - Pb3 (po4)2-precipitation was used to study the ATPase activities both in fertile and sterile anthers of rice (Oryza sativa L. cv. Marxie). At big-vacuole pollen stage, tapetum of the fertile anthers showed high ATPase activity in their nuclei. In fertile pollen, ATPase was localized on the outside surface of the exine and in the nucleus both at big-vacuole and at bi-nucleate pollen stage. At late bi-nucleate pollen stage, a large amount of Pb3 (PO4)2 precipitated in endintine of the fertile pollen. In sterile anthers, tapetum was fully degenerated at big-vacuole pollen stage. In sterile pollen, ATPase was localized both on the plasmamembrane and in intine. These phenomena lasted to the bi nucleate pollen stage. In addition, most of the sterile pollen did not show well developed endintine. Based on the above results, we suggested that abortive tapetum could not provide enough nutrition for pollen development, and the high ATPase activity both on plasma membrane and in intine would likely result in ATP shortage in sterile anthers. PMID- 12549214 TI - [Study on in vitro culture and cryopreservation by vitrification of blastocysts derived from single mouse 2-cell embryos' blastomeres]. AB - The present experiments were designed to study the effects of glucose, EDTA, glutamine on the in vitro development of single blastomeres from 2-cell embryos in mouse, and the efficiency of cryopreservation of blastocysts from single blastomers with different vitrification. Single blastomeres derived from female ICR x male BDF1 2-cell embryos were cultured in mKRB with or without glucose, EDTA and glutamine, respectively. The expanded blastocyst rates were significantly different between in mKRB with glucose and without glucose (34% vs 65%); The blastomeres were cultured in mKRB with EDTA and glutamine but glucose, the expanded blastocyst rate (90%) was significantly higher than other groups. The blastocysts derived from single blastomeres were vitrified in liquid nitrogen after equilibration in GFS40 for 0.5-2 min, the survival rate 24%-51%. The blastocysts were pretreated in mPBS with 10% glycerol for 5 min, followed by exposure to GFS40 at 25 degrees C for 0.5 min, then vitrified in liquid nitrogen(two-step method), the survival rate was 61%. However, the survival rates increased to 64% and 70% when the blastocysts were vitrified(one-step method) ater equilibration in EFS40 at 25 degrees C for 0.5-1 min. PMID- 12549215 TI - [Rapid induction of senescence-like changes in human umbilic vein endothelial cells(HUVECs) by C6 ceramide]. AB - Ceramide, a key molecule in sphingolipid metabolism and a candidate second messenger, has been shown to inhibit the activity of phospholipase D. This biochemical pathway has been implicated to regulate cell differentiation, apoptosis and cellular senescence. Ceramide is generated in response to a number of extracellular inducers(for example: TNF, IL-1 and Fas ligands etc.), and acts as a second messenger to mediate many of the effects of these inducers. HUVECs are the monolayer cells located inside the vein wall and play an important role in the regulation of vein physiology and blood function. It has been reported that the C6 ceramide can induce senescence of WI-38 HDF and promote the activity of beta-galactosidase, but, C2 ceramide has no such effect. In this study, we investigated the role of C6 ceramide in the senescence of HUVECs. 10 mumol/ml of C6 ceramide treatment for more than 72 hours can induce morphological alterations (such as: enlarged, flattened and irregular cell body), cell cycle arrested at G1 phase and the expression of the senescent histochemical marker-beta-galactosidase in HUVECs. These results showed that C6 ceramide could induce senescence-like changes of HUVECs. The detection of reactive oxygen species(ROS) and the anti oxidative ability of the cells showed that the C6 ceramide induced senescence like cells still have normal ability of anti-oxidation. Further investigations are ongoing. PMID- 12549216 TI - [Detection and analysis of alien genes in transgenic rice by fluorescence in situ hybridization]. AB - Using fluorescence in situ hybridization (FISH) with metaphase preparations, we localized a transferred barnase-psl DNA sequence onto chromosomes in 8 rice transgenic plants. All the tested rice transgenic lines showed hybridization signals on the middle and terminal regions of chromosome arms except for those close to centromeres. In two lines, two different integration sites were identified, and the other lines showed only one integration site. With the aid of Southern analysis and expression detection, we found that the barnase tended to show a higher level expression in the lines whose integration sites near the distal regions of chromosomes, while the expression level became lower in the lines whose integration sites near the centromeres. This result suggested a possible relationship between chromosomal location of transgenes and the expression level. However it showed no obvious relationship between copy numbers and expression levels. In most cases, the results of multi-color FISH showed that barnase-ps1 always integrated at the same position on the chrmosome as the reporter genes(pHctinG). PMID- 12549217 TI - [Studies on genetic diversity in gonochoristic offspring produced from mating between two different gynogenetic clones of silver crucian carp]. AB - Based on the discovery of gonochoristic reproductive mode in silver crucian carp (Carassius auratus gibelio), 18 individuals from the mated offspring between clone F and clone D and their parents were analyzed using 11 RAPD primers. The mated offspring differentiated into three phenotypes. One phenotype is similar to that of clone F (SF). The second is similar to that of clone D (SD). The third shows a novel longer and thinner body type (NL). Electrophoretogram results show abundant polymorhic DNA fragments among individuals in the FD mated offspring. Obviously, the polymorphic DNA fragments originate from the genome recombination owing to the gonochoristic reproductive mode. The FD mated offspring amplify different fingerprints from maternal, and their fingerprints are different from each other. These different DNA fragments can be divided into four groups according to their origin. The average genetic distance among individuals from the FD offspring (0.23 +/- 0.123) is much higher than that of allogynogenetic offspring, which is only about 0.01. Of the three phenotypes in the FD offspring, the average distances among SD is the highest (0.235 +/- 0.097). The next is SF phenotype, in which the average distances is 0.148 +/- 0.073. The average distances among NL is the lowest (0.094 +/- 0.083). The gonochoristic reproductive mode introduces high genotypic variability and underlines genetic diversity. A dendrogram was constructed by NJTREE cluster analysis based on a total of 2320 distinguishable fragments (116 per individual). NL3 and maternal are most closely related, and SD3 and paternal are most closely related. The average genetic distances between SD and SF or NL (> 0.31) is higher than that of between SF and NL (0.124). Consistent with previous study, the genetic distance between maternal and paternal is 0.35. The average genetic distances between FD offspring and parents are both about 0. 32. But the similarities among the three phenotypes in mated group FD offspring and parents are different. The similarity between SF and maternal (71%) is higher than similarities among other phenotypes and maternal. The similarity between SD and paternal (70%) is higher than similarities among other phenotypes and paternal. The results suggest the similarities among three phenotypes in the FD offspring and parents are related with their phenotypes because some DNA fragments specific for phenotypes were amplified in this study. The revelation about reproductive diversity will be able to open the door in which complicated mechanism has been locked in unisexual organisms for long time, and establish the important roles of silver crucian carp in the studies on evolutionary genetics in unisexual or polyploid vertebrates. PMID- 12549218 TI - [The fungicidal effect of the antibacterial peptide CM4 from Chinese silkworm on Fusaricum moniliforme]. AB - The antibacterial peptide CM4 inhibits growth of conidia of Fusaricum moniliforme. The observation by using the electron microscope showed that some cells were empty and interior structure twisted into helix. Conidia cells in control group were in good condition, normally grew up. The fungal conidia treated with the antibacterial peptide CM4, then confocal laser scanning microscope were performed. As a result, it was showed that FITC-labeled peptide assembled in cells, centre of which had more peptides than two ends. Then conidia split, became empty. Finally the cells plasma membrane disintegrated and died, lead to small fragments. PMID- 12549219 TI - [Effects of the physical configuration of plasmid DNA and other factors on production of fertile transgenic wheat plants]. AB - The applicability of hyperosmotic treatment and different configurations of plasmid DNA for stable transformation of wheat mediated by particle bombardment was investigated. Hyperosmotic treatment increased the frequency of transient expression and had also a positive effect on stable transformation. Denaturation of plasmid DNA prior to bombardment led to dramatic reduction of transient expression. However, there were no marked differences between single-stranded and double-stranded DNA in stable transformation. Single-stranded plasmid DNA, double stranded plasmid DNA in linear state and double-stranded plasmid DNA in circular state could all be used to produce transgenic wheat plants. A total of 26 independent transgenic plants of winter wheat genotype Florida and 4 independent transgenic plants of spring wheat genotype Veery were obtained. Most transgenic plants have set seeds. T2 seeds of some spring wheat transgenic plants have also been harvested. PMID- 12549220 TI - Effects of polyamines on organogenesis and somatic embryogenesis of Lycium barbarum calli. AB - Levels of three endogenous free polyamines (PAs), Put, Spd and Spm, were detected during organogenesis and somatic embryogenesis (SE) of Lycium barabrum (L.) calli. The predominant forms of PAs in organogenesis and SE were found to be Put and Spd, respectively. In both developmental pathways, the changes of Put content were very similar, i.e., it accumulated quickly in the initial stages of calli differentiation and then decreased; with the further morphogenesis, increase in Put level was also observed. The highest level of Spd was obtained at day 1 of calli organogenesis; while, after 1 day of culture for calli SE, Spd level began to increase and reached a maximum at day 10. The treatments with exogenous PAs had positive effects on both organogenesis and SE, especially those of Spd on organogenesis and SE and Put on SE. However, Put showed no effects on adventitious bud formation. CHA, which inhibits the activity of Spd synthetase, prevented both adventitious bud formation and further development of somatic embryo into plantlet. Although MGBG, a specific inhibitor of S-adenosyl methionine decarboxylases, had little effects on organogenesis, it reduced the number of somatic embryo and the plantlets subsequently regenerated. Such inhibitions could be reversed by Spd (50 mumol/L). These results from organogenesis and SE of the same specie indicate that PAs influenced these two in vitro morphogenesis pathways. PMID- 12549221 TI - [Effects of short interval successive partial hepatectomy on ACP, AKP, HSC70/HSP68 and PCNA in rat liver]. AB - In this paper, the models of 36-4-4 SISPH and 4-36-36-36 SISPH were used to analyze the changes of activity and content of ACP, AKP, HSC70/HSP68 and PCNA in rat liver. The results showed that the activities of 140 kD ACP and AKP in SISPH were increased following the increase of SISPH number of times, but that of 160 kD ACP and AKP were decreased following the increase of SISPH number of times. The content of PCNA in 4-36-36-36 SISPH were more than that in 36-4-4 SISPH, in contrast for HSC70/HSP68 in these two models. Therefore, the content and activities of ACP, AKP, HSC70/HSP68 and PCNA could be strongly effected by SISPH number of times and SISPH methods. Its mechanisms and physiological significance were discussed. PMID- 12549222 TI - [Epstein-Barr virus latent membrane protein 1 induces the telomerase activity in human nasopharyngeal epithelial cells]. AB - Telomerase activation has been linked to cell immortalization in vitro and tumorigenicity in vivo. In this study, for the first, we reported that Epstein Barr virus activated the telomerase activity of human nasopharyngeal epithelial cells in the early stage of immortalization as tested by the PCR-ELISA. The telomerase activity in nasopharyngeal epithelial cells was only observed in presenescent cells. It was implicated that Epstein-Barr virus induced the escape of nasopharyngeal epithelial cells from senescence via the activation of telomerase. We further showed that telomerase activation in infected cells was dependent on the protein level of latent membrane protein 1 (LMP1) encoded by Epstein-Barr virus using a Tetracycline regulatory cell line expressing LMP1, pTet-on-LMP1-HNE2. The activity of telomerase in nasopharyngeal cells was decreased when the protein level of LMP1 was blocked by antisense LMP1 plasmid DNA. And the activity of telmerase was also related to the carboxyl terminus of LMP1. It was implicated that the ability of Epstein-Barr virus to suppress senescence is associated with telomerase activation by LMP1. PMID- 12549223 TI - [Telomerase activity inhibition and apoptosis induction of BEL-7404 human hepatoma cells by antisense oligonucleotides to telomerase RNA component]. AB - Telomerase activity was detected in all of four human hepatoma cells but absent in normal liver tissue. Telomerase activity of BEL-7404 human hepatoma cells was inhibited effectively by antisense oligonucleotide to telomerase RNA component at final concentration of 1 mumol/L, whereas sense and missense oligonucleotides have no effects on its activity. The inhibition of telomerase activity was weakened, as the concentration of antisense oligonucleotide decreased. Treating BEL-7404 human hepatoma cells 96 hours continuously by the antisense oligomers at final concentration of 5 mumol/L, the morphology of treated cells changed and apoptosis percent of the cells increased markedly. PMID- 12549225 TI - [A bicistronic retroviral vector containing MGMT and MDR1 drug resistance genes transfer into human umbilical cord blood CD34+ cells to improve combination chemotherapy tolerance]. AB - To explore whether human umbilical cord blood hematopoietic progenitor cells transduced with human O6-methylguanine-DNA-methyltransferase (MGMT) and multidrug resistance gene (MDR1) increase resistance to 1,3-Bis(2-Chloroethy1)-1 Nitrosourea (BCNU) and P-glycoprotein effluxed drugs, the present authors obtained a full length cDNA fragment encoding MGMT from liver tissue of a patient with cholelithiasis by RT-PCR. A bicistronic retroviral vector G1Na-MGMT-IRES MDR1 cDNA was constructed and transfected the packaging cell lines GP + E86 and PA317 by electric performation method, using the medium containing VCR and BCNU for cloning selection and ping-ponging supernatant infection between ecotropic producer clone and amphotropic producer clone, cord blood CD34+ cells were enriched with a high-gradient magnetic cell sorting system (MACS), and then transfected repeatedly with supernatant of retrovirus containing human MGMT and MDR1cDNA under stimulation of hemapoietic growth factors. PCR, RT-PCR, Southern blot, Northern blot, Western blot, FACS and MTT assay were used to evaluate the transfer and expression of the double genes in cord blood CD34+ cells. The cDNA encoding MGMT was verified by DNA sequencing and the bicistronic retroviral vector was confirmed by restriction endonuclease analysis. The purity of cord blood CD34+ cells was approximately 92% and recover rate was 75%, the highest titer of recombinant amphotropic retrovirus in the supernatant was up to 5.8 x 10(5) cfu/ml. The efficiency of gene transduction was 18% and 20% tested by colony formation and PCR, respectively. No helper virus was found by both nested PCR and rescue assay. The results showed that dual drug resistance genes have been integrated into the genomic DNA of cord blood CD34+ cells and expressed efficiently. The MTT analysis showed a 4.5 to 7.8-fold increase of resistance of transducted cells to BCNU and P-glycoprotein effluxed drug as compared with the nontransduced cells. This study provided a foundation for ameliorating combination chemotherapy toxicity in tumor clinical trial. PMID- 12549224 TI - [GnT-V overexpression in human hepatocarcinoma cells affects its migration and expression of cell adhesion molecules]. AB - The purpose of this paper is to study the effect of N acetylglucosaminyltransferase V (GnT-V) overexpression on the migration of 7721 cells and its mechanism. The abilities of migration of both 7721 cells transfected with GnT-V cDNA and 7721 cells transfected with pcDNA3 was detected, the expressions of integrin and E-cadherin which are important adhesion molecules on surface membrane and closely related to the abilities of invasion and metastasis. Cell migration abilities were measured by the agarose drop explant method. Flow cytometric analysis (FACS) was applied to determine the relative amounts of integrin alpha 5 and beta 1 subunits on the cell surface while RTPCR was carried out to determine the expression of their mRNA. The expression of E cadherin was examined by the immunocytochemical ABC method. Western blot analysis was carried out to examine the expression of beta-catenin. GnT-V overexpression enhanced evidently the migration ability of 7721 cells and increased the amount of integrin alpha 5 subunit to 2.9 times of that of control while the amount of beta 1 subunits was not significantly changed. Besides, the expressions of E cadherin and beta-catenin were enhanced at different levels in GnT-V/7721 cells compared with mocked. The results suggested that the overexpression of GnT-V related to the production of N-linked sugar chains could promote the expressions of integrin, E-cadherin and beta-catenin on 7721 cells so that the migration ability of tumor cells was enhanced. PMID- 12549226 TI - [Histochemical localization and quantitative analysis of anthraquinones in rhizome of Polygonum cuspidatum]. AB - Anthraquinones were histochemically locayed and content were determined in rhizome of Polygonum cuspidatum. Anthraquinones were located in the parenchyma cells of rhizome, including vascular ray, part of parenchyma cell in phloem, cortex and pith. Phelloderm and ray initial also accumulated a small amount anthraquinones. The content of total anthraquinones was highest in pith that consisted of parenchyma cells, and higher in bark (including phloem, cortex and periderm) than in xylem. The content of total anthraquinones was highest in three year-old rhizome, and higher in biennial rhizome than in annual rhizome. It suggests that correlation exist between accumulation of anthraquinones and the growth age of rhizome. PMID- 12549227 TI - [Relationship of hR24L gene expression, DNA damage repair and G2/M phase cell cycle arrest with ion radiation]. AB - The hR24L gene ORF was cloned from the total RNA of HeLa cells by RT-PCR method. There is no mutation of the hR24L gene in HeLa cells. Ion radiation significantly increased the expression of hR24L gene, and the sense hR24L enhanced and accelerated the cell cycle arrest at G2/M phase in HeLa cells. Besides, it should seem that the overexpression of hR24L gene could enhance the repair ability of DNA damage induced by ion radiation, and vice versa. PMID- 12549228 TI - [Establishment of human neuroectodermal tumor cell line (PFSK) with stable expression of xanthine oxidoreductase]. AB - Xanthine Oxidoreductase (XOR) is the key enzyme in purine metabolism and also produces oxygen free radicals. As a continuation of our previous work, in this study, we constructed a retrovirus expression vector (pLNCX2-XOR) containing full length of XOR cDNA. Retrovirus was produced by the virus package cell line PT-67 following the transfection of PT-67 with pLNCX2-XOR and used to infect the human primitive neuroectodermal tumor cell line (PFSK). Infected PFSK cells were selected by G418 to establish cell line with stable expression of XOR. The expression of XOR in the cell line we established was confirmed by RT-PCR, Immunocytochemistry and XOR activity assay. PMID- 12549229 TI - [The comparative study on ova superovulated, in vitro fertilization and rate of pregnancy at different ages]. AB - The development of oocytes, superovulated at 28, 56 and 112 days in three mice strains (ICR, B6C3F1, and C57BL/6J) with PMSG and HCG, were examined using the techniques of in vitro fertilization, culture, and transfer of these two-cell embryos to pseudopregnant recipients. The highest numbers of oocytes were obtained from superovulated 28-day-old mice in three strains. Approximately 90% of oocytes developed to the two-cell stage after in vitro fertilization, and about 50% became pregnant through the recipients. These results suggested that donor age at 28 days had prominent discrepancy with 56 and 112-day-old mice (P < 0.01) in oocytes superovulation, and no influence on the rate of insemination and pregnancy. PMID- 12549230 TI - Comparative MRI compatibility of 316 L stainless steel alloy and nickel-titanium alloy stents. AB - The initial success of coronary stenting is leading to a proliferation in peripheral stenting. A significant portion of the stents used in a clinical setting are made of 316 low carbon stainless steel (SS). Other alloys that have been used for stent manufacture include tantalum, MP35N, and nickel-titanium (NiTi). The ferromagnetic properties of SS cause the production of artifacts in magnetic resonance imaging (MRI). The NiTi alloys, in addition to being known for their shape memory or superelastic properties, have been shown to exhibit reduced interference in MRI. Thus, the objective of this study was to determine the comparative MRI compatibility of SS and NiTi stents. Both gradient echo and spin echo images were obtained at 1.5 and 4.1 T field strengths. The imaging of stents of identical geometry but differing compositions permitted the quantification of artifacts produced due to device composition by normalizing the radio frequency shielding effects. These images were analyzed for magnitude and spatial extent of signal loss within the lumen and outside the stent. B1 mapping was used to quantify the attenuation throughout the image. The SS stent caused significant signal loss and did not allow for visibility of the lumen. However, the NiTi stent caused only minor artifacting and even allowed for visualization of the signal from within the lumen. In addition, adjustments to the flip angle of standard imaging protocols were shown to improve the quality of signal from within the lumen. PMID- 12549231 TI - Real-time volume rendered MRI for interventional guidance. AB - Volume renderings from magnetic resonance imaging data can be created and displayed in real-time with user interactivity. This can provide continuous 3D feedback to assist in guiding an interventional procedure. A system is presented which can produce real-time volume renderings from 2D multi-slice or 3D MR pulse sequences. Imaging frame rates up to 30 per second have been demonstrated with a latency of approximately one-third of a second, depending on the image matrix size. Several interactive capabilities have been implemented to enhance visualization such as cut planes, individual channel scaling and color highlighting, view sharing, saturation preparation, complex subtraction, gating control, and choice of alpha blending or MIP rendering. The system is described and some interventional application examples are shown. To view movies of some of the examples, enter the following address into a web browser: http://nhlbi.nih.gov/labs/papers/lce/guttman/rtvolmri/index/htm. PMID- 12549232 TI - Normal systolic and diastolic functions of the left ventricle and left atrium by cine magnetic resonance imaging. AB - Volume and phase characteristics of the left ventricle (LV) and left atrium (LA) were assessed in 31 healthy Asian adults (19 males and 12 females) using cine magnetic resonance imaging (MRI) and an automated boundary detection algorithm. Volume indexes of the LV and LA were smaller than published results obtained mostly from Westerners. Other than LV mass index and percent emptying of the LA, there was no gender difference in all LV/LA indexes. In associating LV/LA functions with the body size and heart rate (HR), we found that LV mass and the minimum LA volume correlated strongly with the body surface area, the maximum LA volume and the reservoir volume correlated strongly with the body weight, and the time to LV peak-filling rate (LVPFRt) and the time to LA peak-emptying rate (LAPERt) correlated strongly with the HR. In associating LV with LA functions, we found that LA conduit volume contributed more than 50% of the LV stroke volume, and correlated with both systolic and diastolic functions of the LV. Moreover, LVPFRt and LAPERt were virtually identical, indicating a mechanical coupling between LV and LA during diastole. In conclusion, using time-resolved, three dimensional volume data obtained from cine MRI, we have established normative values of LV and LA functions and their functional relationships in healthy Asian adults. The imaging acquisition protocol, data analysis algorithms, and the established normative values provide the basis for the study of left heart function in patients. PMID- 12549233 TI - Fully automated registration and warping of contrast-enhanced first-pass perfusion images. AB - Respiratory motion during acquisition of first-pass myocardial perfusion images results in translation, distortion from out-of-plane motion, and changes in left ventricular geometry. Together these effects make visual image analysis more difficult and limit methods of quantitative analysis of contrast kinetics. We present a fully automated registration and warping algorithm for correcting translation and geometric distortions using a statistically based image registration method. Twelve patients (mean age 51 +/- 12 years) were studied 3 +/ 1 days after reperfused first myocardial infarction. Perfusion images were acquired during bolus administration of nonionic Gd-DTPA. Pixel intensity statistics were computed for each image in the neighborhood of high spatial frequencies. These statistics were then used to register and warp each target image (image to be registered and warped) to a common template image. Average image-to-image vertical translation was 2.6 +/- 0.8 pixels (3.4 +/- 1.0 mm) prior to processing and 0.9 +/- 0.3 pixels (1.2 +/- 0.4 mm) post-processing (P < 0.0001). Mean image-to-image horizontal translation was 1.7 +/- 1.2 pixels (1.8 +/- 1.2 mm) before and 1.3 +/- 0.7 pixels (1.4 +/- 0.7 mm) after processing (P = 0.05). Left ventricular endocardial area varied an average of 105 +/- 55 pixels (140.7 +/- 53.7 mm2) between images prior to processing vs. 51 +/- 15 pixels (68.3 +/- 20.1 mm2) after processing (P < 0.001). Thus automated, statistically based registration and warping of perfusion images is effective in reducing image to-image translation. This method may permit more sensitive qualitative and quantitative evaluation of myocardial contrast-enhanced first-pass images. PMID- 12549234 TI - Dobutamine induced myocardial perfusion reserve index with cardiovascular MR in patients with coronary artery disease. AB - Currently, adenosine or dipyridamole is commonly used for the assessment of perfusion reserve. With intolerance to these agents, dobutamine can be used alternatively or it can be used for a combined examination of wall motion and perfusion. The aim of the study was to analyze the feasibility of cardiovascular magnetic resonance (CMR) to assess perfusion reserve with dobutamine. Alterations of myocardial perfusion were noninvasively assessed in 23 patients with and 4 without significant coronary artery disease by calculation of a myocardial perfusion reserve index from the upslope of the signal intensity curves of a first pass gadolinium bolus before and during dobutamine infusion (20 micrograms/min/kg). An ischemic threshold value of perfusion reserve index was determined from patients without significant coronary artery disease. Significant differences were found between ischemic and remote to ischemic segments in patients with single vessel disease (0.90 +/- 0.18 vs. 1.73 +/- 0.32, p < 0.0001). Differences between nonischemic segments in patients without and ischemic segments in patients with coronary artery disease were significant (2.0 +/- 0.39 vs. 0.97 +/- 0.20, p < 0.001). A cut-off value for myocardial perfusion reserve index of 1.22 for the detection of significant coronary artery stenosis yielded a sensitivity, specificity, and diagnostic accuracy of 81, 73, and 77%, respectively. Dobutamine MR is feasible in the evaluation of myocardial perfusion and can be used for the detection of myocardial ischemia alternatively to adenosine or dipyridamole in patients with coronary artery disease. PMID- 12549235 TI - The clinical significance of aortic compliance and its assessment with magnetic resonance imaging. AB - The biophysical properties of the aortic wall seems to play a significant role in the pathogenesis of cardiovascular disease such as atherosclerosis, hypertension, aneurysm formation, Marfan's syndrome, and in normal aging. The presence and the proportion of smooth muscle, collagen, and elastin proteins contribute to the compliance of the vessel wall with the latter being the most extensible component. However, elastin fibers fracture at low stresses contributing to a decrease of the aortic compliance and consequently to an elevation of the pulse pressure, which is a risk factor of cardiovascular disease. Early detection of a decrease in the aortic compliance could help to identify early cardiovascular disease in asymptomatic patients and monitor the results of the therapeutic interventions. Therefore, estimation of the aortic compliance can be used for both screening as well as long-term follow-up. Magnetic resonance imaging which is a noninvasive, accurate, and reproducible method can estimate the compliance of the aortic wall either by measuring the relative change in cross sectional area of a chosen segment using ECG-triggered spin echo or gradient echo sequences or by measuring the pulse wave velocity through the aorta using the phase contrast-magnetic resonance imaging (PC-MRI) technique. Both techniques have been validated and many sudies suggest MRI as a valuable tool for evaluating aortic wall function. However, large prospective studies are mandatory for the method to be established as a screening tool. PMID- 12549236 TI - Decreased high-energy phosphate ratios in the myocardium of men with diabetes mellitus type I. AB - AIMS/HYPOTHESIS: To investigate whether alterations in high-energy phosphates occur in the myocardium of persons with diabetes mellitus type I. Microvascular abnormalities and dysfunction via thickening of the basement membrane are known to occur in diabetic patients. Myocardial high-energy phosphates have been shown to be reduced by ischemia, and alterations of the cardiac metabolism are the primary consequence of myocardial ischemia. METHODS: The present study involved 34 male patients (mean age 35.5 +/- 10.1) with diabetes mellitus type I and 35 healthy male volunteers (mean age 36 +/- 8.6) as age-matched controls. Phosphorus 31 magnetic resonance spectroscopic imaging of the heart was performed in all subjects using a 1.5-T whole-body magnetic resonance scanner. The ratios of phosphocreatine (PCr) to beta-adenosinetriphosphate (beta-ATP) were calculated. Moreover, echocardiographic evaluation and stress tests were performed in all individuals. RESULTS: The myocardium of patients with diabetes mellitus type I showed significantly decreased ratios of PCr to beta-ATP compared with healthy controls in the left ventricle (1.90 +/- 0.4 vs. 2.15 +/- 0.3, p < 0.05). We found a moderate negative correlation between the ratio of PCr to beta-ATP in the left ventricle and both, the diastolic left ventricular function (E/A; r = -0.41) and the glycohemoglobin A1c (GHbA1c; r = -0.42). CONCLUSION: This study demonstrates for the first time a decreased ratio of PCr to beta-ATP in the myocardium of persons with diabetes mellitus type I without a known history of coronary heart disease. PMID- 12549237 TI - Diagnosis of a postsurgical aortic pseudoaneurysm in Shone syndrome by cardiovascular magnetic resonance. PMID- 12549238 TI - Ventricular volume and mass by CMR. PMID- 12549240 TI - Agricultural behavioral health. PMID- 12549239 TI - Report from the First International Conjoint Conference on Cardiovascular Magnetic Resonance and Echocardiography. AB - The opinions formulated in this conference represent expressions of promise and enthusiasm about charting the course for the future of cardiovascular imaging and the role that might be played by CMR and echocardiography. All attendees agreed that this conference provided a novel venue for friendly debate and discussion in an open forum that could facilitate research and clinical service planning, and that other similar conferences should be held to focus on additional methodologies and topics. Indeed, planning is already underway for a Second Conference with comparative CMR and nuclear imaging as a focus. PMID- 12549241 TI - Temporal variation of indoor air quality in an enclosed swine confinement building. AB - Human health hazards can exist in swine confinement buildings due to poor indoor air quality (IAQ). During this study, airborne dust and ammonia concentrations were monitored within a working farrowing facility as indicators of IAQ. The purposes of this study were to assess the temporal variability of the airborne dust and ammonia levels over both a daily and seasonal basis, and to determine the accuracy of real-time sensors relative to actively sampled data. An ammonia sensor, aerosol photometer, indoor relative humidity sensor, and datalogger containing an indoor temperature sensor were mounted on a board 180 cm above the floor in the center of a room in the facility. Sensor readings were taken once every 4 minutes during animal occupancy (3-week intervals). Measurements of total and respirable dust concentrations by standard method, aerosol size distribution, and ammonia concentrations were taken once per week, in addition to temperature and relative humidity measurements using a thermometer and sling psychrometer, respectively. Samples were taken between September 1999 and August 2000. Diurnal variations in airborne dust revealed an inverse relationship with changes in indoor temperature and, by association, changes in airflow rate. Ammonia levels changed despite relatively stable internal temperatures. This change may be related to both changes in flow rates and in volatility rates. As expected, contaminant concentrations increased during the cold weather months, but these differences were not significantly different from other seasons. However, total dust concentrations were very low (geometric mean = 0.8 mg/m3) throughout the year. Likewise, ammonia concentrations averaged only 3.6 ppm in the well maintained study site. PMID- 12549242 TI - Quantitative exposure assessment for the combustion of meat and bone meal derived from specified risk material in the context of BSE in Ireland. AB - The probability and severity of an adverse event can be analyzed by quantitative exposure assessment (QEA). This methodology was applied to model the human health risks associated with the combustion of specified risk material (SRM) derived meat and bone meal (MBM) in a combustion facility. The identification of MBM and SRM as significant factors in the spread of bovine spongiform encephalopathy (BSE) has resulted in restrictions on their use and movement, and this has led to a requirement for alternative end-uses for these products. A stochastic (Latin Hypercube sampling) simulation model was developed to assess the exposure and hence the risks associated with the use of SRM-derived MBM in a combustion facility. The model simulates the potential infectivity pathways that SRM-derived MBM follows, including its production from animals potentially infected with sub clinical BSE and subsequent processing of the material with segregation and heat treatments. A failure probability was included to take account of sub-optimal operating conditions. Two scenarios, reflecting the infectivity risk in different animal tissues as defined by the European Commission's scientific steering committee (SSC), were performed with 100,000 iterations of the model. Model results showed that the societal exposure to humans resulting from the combustion of SRM-derived MBM is extremely small (mean values ranging from 7.57 x 10(-6) ID50/year to 8.38 x 10(-5) ID50/year). The resulting societal risks are significantly less than the background societal risk of approximately 2.5 cases of sporadic CJD in Ireland each year. A sensitivity analysis revealed that the species barrier had a large impact on exposure calculations and hence should be the focus of further scientific investigation to reduce our uncertainty about this parameter. The model predicts that material spillage into untreated effluent represents the biggest risk to humans, indicating that efforts for risk mitigation should be focused on reducing the potential for spillage. PMID- 12549243 TI - Agricultural machinery safety alert system using ultrasonic sensors. AB - This article introduces a conceptual safety alert system using ultrasonic sensors. The safety alert system was designed to detect moving objects in the vicinity of agricultural machinery. This system uses two ultrasonic sensors to detect the distances between the sensors and the moving object and a position detection algorithm to determine the moving object's position relative to the machinery. A stationary test bench was built to prove the concept of the safety sensing system. Validation tests in an outdoor environment indicated that the conceptual safety alert system was capable of detecting the position of a moving object in the vicinity of agricultural machinery in real time, and generating a timely warning signal to raise the attention of the operator for ensuring safe operations. This result proved that the conceptual system has tremendous potential for agricultural machinery applications. PMID- 12549244 TI - Pesticide knowledge and risk perception among adolescent Latino farmworkers. AB - A substantial proportion of the agricultural production in the U.S. is dependent on the labor of Latino farmworkers. While exact figures are not known, it is estimated that adolescents make up 7% of this valuable workforce. These young workers may be at increased risk for the toxic effects of environmental exposures encountered during their work. Furthermore, language barriers and health beliefs may influence the risk perceptions of this population. We conducted a cross sectional survey of migrant adolescent farmworkers in 1998 to investigate their work practices, health beliefs, and pesticide knowledge. The large majority of the adolescents in our sample were from Mexico, and 36.3% spoke primarily indigenous languages. Many of the adolescents (64.7%) were traveling and working in the U.S. independent of their parents. Few of the adolescents reported having received pesticide training; however, 21.6% of the sample reported that their current work involved mixing and/or applying agricultural chemicals. The scores on the pesticide knowledge questionnaire were found to significantly predict self reported use of protection for adolescent farmworkers. The results of this study point to a need for improved pesticide training in youth agricultural workers and specialized education efforts directed toward minorities who speak indigenous dialects. Special attention is merited toward adolescent farmworkers who report that their work includes mixing or applying agricultural chemicals. As the number of adolescent farmworkers increases in the U.S. and the characteristics of the migrant stream continue to change, culturally and developmentally appropriate instruments are needed to adequately assess the health beliefs and protective practices of this population. PMID- 12549245 TI - Development and evaluation of the Kids Count Farm Safety Lesson. AB - The Kids Count Farm Safety Lesson was delivered to nearly 2,000 fifth-grade students in 15 rural schools in Hillsborough County, Florida. The lesson covered animal, machinery, water, and general safety topics applicable to farming in Florida. A staggered pretest-posttest study design was followed whereby five schools received a multiple-choice pretest and posttest and the remainder of the schools (N = 10) received the posttest only. Results of the study showed a significant increase in the mean number of correct answers on the posttests compared to the pretests. There was no significant difference in the mean number of correct answers of those students who received the pretest and those students who had not, eliminating a "pretest" effect. This study fills an important gap in the literature by evaluating a farm safety curriculum offered in the elementary school setting. It also included migrant schoolchildren in the study population. It is strongly recommended that agricultural safety information be included into the health education curriculum of these elementary schools. PMID- 12549246 TI - Pesticide use and practices in an Iowa farm family pesticide exposure study. AB - Residents of Iowa were enrolled in a study investigating differences in pesticide contamination and exposure factors between 25 farm homes and 25 non-farm homes. The target pesticides investigated were atrazine, metolachlor, acetochlor, alachlor, 2,4-D, glyphosate, and chlorpyrifos; all were applied to either corn or soybean crops. A questionnaire was administered to all participants to determine residential pesticide use in and around the home. In addition, a questionnaire was administered to the farmers to determine the agricultural pesticides they used on the farm and their application practices. Non-agricultural pesticides were used more in and around farm homes than non-farm homes. Atrazine was the agricultural pesticide used most by farmers. Most farmers applied pesticides themselves but only 10 (59%) used tractors with enclosed cabs, and they typically wore little personal protective equipment (PPE). On almost every farm, more than one agricultural pesticide was applied. Corn was grown by 23 (92%) farmers and soybeans by 12 (48%) farmers. Of these, 10 (40%) grew both soybeans and corn, with only 2 (8%) growing only soybeans and 13 (52%) growing only corn. The majority of farmers changed from their work clothes and shoes in the home, and when they changed outside or in the garage, they usually brought their clothes and shoes inside. Applying pesticides using tractors with open cabs, not wearing PPE, and changing from work clothes in the home may increase pesticide exposure and contamination. Almost half of the 66 farm children less than 16 years of age were engaged in some form of farm chores, with 6 (9%) potentially directly exposed to pesticides, while only 2 (4%) of the 52 non-farm children less than 16 years of age had farm chores, and none were directly exposed to pesticides. Farm homes may be contaminated with pesticides in several ways, resulting in potentially more contamination than non-farm homes, and farm children may be directly exposed to pesticides through farm chores involving pesticides. In addition to providing a description of pesticide use, the data presented here will be useful in evaluating potential contributing factors to household pesticide contamination and family exposure. PMID- 12549247 TI - [Method of treatment of purulent wounds]. PMID- 12549248 TI - [The assessment of wound healing action of flumexid, a modern home medicinal agent]. PMID- 12549249 TI - [Cryotherapy of purulent wound]. PMID- 12549250 TI - [Aspects of open plasty for the abdominal wall hernia in patients with excessive body mass and morbid obesity]. PMID- 12549251 TI - [Comparative evaluation of contemporary methods for inguinal canal open plasty using synthetic prosthesis]. PMID- 12549252 TI - [Aspects of wound healing in implantation hernioplasty]. PMID- 12549254 TI - [Internal organs protection in the "open" surgery in patients with intra abdominal purulent-septic diseases]. PMID- 12549253 TI - [Experimental substantiation of application of hydrophilic 10% Propes ointment in stage 1 wounds]. PMID- 12549255 TI - [Morphogenesis of healing in soft tissue purulent, infected wounds as a result of electrochemically activated solution application]. PMID- 12549256 TI - [Use of polyurethane "Bioderm" for the treatment of long lasting wounds]. PMID- 12549257 TI - [Varicose vein surgery complicated by trophic ulcers]. PMID- 12549258 TI - [Surgical wound healing and anastomosis in patients with stomach cancer during immobilized cytostatic treatment]. PMID- 12549259 TI - [Ultrastructural description of processes of reparative regeneration in inter intestinal anastomosis formed by double-row suture method]. PMID- 12549260 TI - [Surgical infected wound treatment in patients with severe gastrointestinal ulcer hemorrhage]. PMID- 12549261 TI - [Aronic oil application for the treatment of wounds and ulcers in surgical practice]. PMID- 12549262 TI - [Combined treatment of patients with diabetic angiopathy of the lower extremities]. PMID- 12549263 TI - [Application of prostaglandin E1 and Naftidrofuryl in combined treatment of purulent-necrotic complications of the diabetic foot syndrome]. PMID- 12549265 TI - [Patogenetic bases of the preparation choice in treatment of purulent wounds]. PMID- 12549264 TI - [Preventive treatment of surgical wound infections in patients with acute surgery related diseases]. PMID- 12549266 TI - [Experience of treatment of purulent wound]. PMID- 12549267 TI - [Treatment and prophylaxis of anastomosis motility disorders after ulcer surgery]. PMID- 12549268 TI - [Morphological investigation of vascular anastomoses]. PMID- 12549269 TI - [Lipid peroxidation in substance dependent patients with purulent-inflammatory diseases of soft tissues]. PMID- 12549270 TI - [Surgical treatment of lower extremity erysipelas]. PMID- 12549271 TI - [Autodermoplasty of the wound defect in patients with diabetes mellitus]. PMID- 12549272 TI - [Prophylaxis of inflammatory complications of wound healing in patients with postoperative hernia of abdominal wall]. PMID- 12549273 TI - [Application of probiotics in the treatment of patients with nonhealing purulent inflammatory wounds]. PMID- 12549274 TI - [Antimicrobic activity and efficacy of application of water-soluble gel Tizol with metallic iodine composition for local treatment of trophic complications of the lower extremities chronic arterial ischemia]. PMID- 12549275 TI - [Invagination anastomosis -- method of choice for the treatment of high small intestinal fistula]. PMID- 12549276 TI - [Closure of the wound defect by transferred flap of skin]. PMID- 12549277 TI - [Significance of ultrasonic reflexotherapy in early preparation of purulent wound for the autodermoplasty]. PMID- 12549278 TI - [Optimization of treatment of patients with acute destructive stagnation mastitis]. PMID- 12549279 TI - [Efficacy of external protection of manual intestinal suture and anastomosis by antibacterial collagen explants in operative treatment of the small intestine and colon fistula]. PMID- 12549280 TI - [Application of the low-intensive laser irradiation for the prophylaxis of the small intestinal anastomosis sutures insufficiency]. PMID- 12549281 TI - [Combined treatment and surgery of extensive purulent necrotic phlegmon]. PMID- 12549282 TI - [Application of gauze with metallic inclusions for the treatment of purulent wounds]. PMID- 12549283 TI - [Minimally invasive surgical procedures for trophic ulcers and distal necrosis in the diabetic foot syndrome]. PMID- 12549284 TI - [Conservative therapy of tissue decubitus in a combined therapy of patients with obliterating disease and diabetes-related vascular injuries]. PMID- 12549285 TI - [Analysis of results of the gender surgical correction in transsexualism]. PMID- 12549286 TI - [Prophylaxis of infectious complications in reconstructive surgery of multifocal atherosclerosis in patients with critical ischemia of organs and tissues]. PMID- 12549287 TI - [Treatment of extensive trophic ulcer of the lower extremities]. PMID- 12549288 TI - [Local injection of fraxiparine for the treatment of the lower extremity trophic ulcer]. PMID- 12549289 TI - [Combined treatment of patients with purulent inflammatory soft tissues by gentaxane, enterosgel and aevit]. PMID- 12549290 TI - [Infection complications of the operative wound healing in laparoscopic surgery]. PMID- 12549291 TI - [Aspects of the knee and foot arterial injuries in patients with the diabetic foot syndrome]. PMID- 12549292 TI - [Application of local prolonged vulnerosorption in prophylaxis of purulent complications of operative wound healing in diffusion peritonitis]. PMID- 12549293 TI - [Comparative evaluation of anastomoses through esophago-intestinal anastomosis]. PMID- 12549294 TI - [Aspects of prophylaxis of the operative wound suppuration in patients with morbid obesity]. PMID- 12549295 TI - [Experience of the medical standards and criteria of quality using ISO 9001:2000 system in surgical departments]. PMID- 12549296 TI - [Prophylaxis and treatment of purulent-septic complications in hematologic patients]. PMID- 12549297 TI - [Cutaneous perfusion pressure as a criterion of chronic critical lower extremity ischemia diagnosis]. PMID- 12549298 TI - [Experimental substantiation for the use of cyanoacrylate adhesive "Katsyl" in abdominal surgery]. PMID- 12549299 TI - [Methods of the efficacy increase in local treatment of purulent wound]. PMID- 12549300 TI - [Prophylaxis of purulent-inflammatory complications during laparoscopic cholecystectomy]. PMID- 12549301 TI - [Changes of the carbohydrate metabolism during combined surgical treatment in patients with diabetes mellitus complicated by necrotic-inflammatory foot injury]. PMID- 12549302 TI - [Principles of surgical treatment of patients with diabetes mellitus complicated by necrotic-inflammatory foot injury]. PMID- 12549303 TI - [Surgical treatment of relapsing inguinal hernia by polypropylene net]. PMID- 12549304 TI - [Comparative characteristics of surgical wound healing which was hermetically closed by conventional methods and electrocautery]. PMID- 12549306 TI - [Healing of the wound after performance of plastic aesthetic intervention]. PMID- 12549305 TI - [Suturing of the spleen wound in its traumatic injury]. PMID- 12549307 TI - [Treatment of severe forms of chronic venous insufficiency: surgical decision of problem of venous trophic ulcers]. PMID- 12549308 TI - [Changes in the homeostasis system under the influence of the peptid bioregulators during the wound healing in experiment]. PMID- 12549309 TI - [The 15-year experience of polysorb application for the treatment of purulent inflammatory wounds]. PMID- 12549310 TI - [Late purulent complications after performance of restoration operation on main vessels]. PMID- 12549311 TI - [Narrow aperture of aorta in combined prosthesis of mitral and aortic valves]. AB - Results of treatment of 520 patients consequently operated on in the clinic for combined mitral-aortal failure were summarized. All the operations were performed using artificial blood circulation and moderate hypothermia (28 ... 32 degrees C). The defense of myocardium was done using medicinal combined ante-retrograde cooling cardioplegia. While performing the analysis there was established the presence of dependency between hospital lethality and the fibrous ring size (FRS) of the aortal aperture (AA). In late follow-up period after the operation the most low frequency of good results was noted in patients with narrow FRSAA (lesser than 24 mm). PMID- 12549312 TI - [Surgical treatment of the double outlet right ventricle in the child's first year of life]. AB - Experience of surgical treatment of the two-barrel branching of main vessels from right ventricle in the first year of life children was summarized. There were operated 105 patients. In 53 (50.5%) infants in arms radical intervention (intraventricular tunneling, operation of Kavashima, arterial switching) was performed with general lethality 24.5%. In 52 (49.5%) patients--palliative operation was done (formation of Blelock--Taussig anastomosis, the pulmonalis artery narrowing) with general lethality 12.7%. PMID- 12549313 TI - [Rationale for antibacterial therapy for acute pancreatitis in postop period]. AB - Results of antibacterial therapy application in 15 patients with an acute pancreatitis (AP) and postoperative period course in 10 were analyzed. Recommendations for application of antibacterial therapy and purposeful sanation in postoperative period were elaborated. PMID- 12549315 TI - [Ways of improvement in surgical treatment of purulent-necrotic complications in the diabetic foot syndrome]. AB - Results of surgical treatment of 168 patients with diabetes mellitus (DM) in the diabetic foot syndrome (DFS) were analyzed. Expediency of operative intervention was substantiated by pathogenic form of affection, its depth and diffusion, composition of microorganisms in purulent-necrotic focus. Performance of more extensive surgical treatment or distal amputation of foot with consequent closure of vast wound defect using full-layer subcutaneous-cutaneous flap with protracted selective intra-arterial anti-aggregate and antibacterial therapy was indicated in order to preserve the lower extremity supporting function and for prophylaxis of diffusion of purulent-necrotic affection. Application of improved tactics and surgical treatment in patients with DFS have allowed to reduce frequency of the lower extremity high amputation performance from 24 to 8.6%. PMID- 12549314 TI - [Modern views on the efficacy of different antibacterial preparations in the treatment and prophylaxis of inflammatory processes in patients after transurethral local ultrasonic hyperthermia of the prostate]. AB - Extensive clinical investigation was accomplished to study the influence of three antibacterial preparations groups (tetracyclines, aminoglycosides and fluorochinolons) on the phagocytic-macrophages and the immune systems in patients, to whom transurethral local microwave hyperthermia (TLMH) of prostate was performed for its benign hyperplasia. The data obtained witness high efficacy of fluorochinolons group preparation (tavanic) toward bacteria of different nosologic groups, as well as to their atypical forms. Pronounced efficacy and simplicity of application of the preparation had allowed to recommend its wide use in urologic practice. PMID- 12549316 TI - [Surgical treatment of cysts and cyst-like formations of pancreas]. AB - Results of treatment of 87 patients with cystic pancreatic formation were analyzed. In 38 patients pancreatic cyst had formatted after destructive pancreatitis, in 28--after pancreatic trauma, in 6--due to virsungolithiasis, in 9--cystadenoma was revealed, in 6--cystadenocarcinoma. In 29 patients operation of internal drainage of cyst was performed. Basing on accumulated experience the authors recommend to perform puncture-cathetherizational intervention under ultrasonic investigation and computeric tomography control, when nonformated or complicated pancreatic cyst is present; in the treatment of uncomplicated nonformated pancreatic cyst the operation of internal drainage, using different organs, stomach, duodenum, small intestine, constitutes the method of choice. When it is impossible to perform puncture-cathetherizational intervention, external drainage of cyst and complex conservative therapy may become the method of choice for the treatment of complicated pancreatic cyst. In the presence of benign cystose tumor it is necessary widely apply radical operation--pancreatic resection. The largest frequency of complications occurrence have been noted after external drainage of cyst, performance of cystogastrostomy, pancreatic resection for cystadenoma and cystadenocarcinoma. The principal of them are: the external pancreatic fistula occurrence (in 9.1% of observations), the cyst recurrency (in 5.7%), erosive hemorrhage (in 4.6%), pancreatitis (in 5.4%), suppuration of postoperative wound (in 8%), stenosis of cystodigestive anastomosis (in 2.3%). Postoperative mortality was 6.8%. PMID- 12549317 TI - [Dynamics of respiratory function in patients operated on lungs]. AB - There were examined 185 patients, operated on lungs, in terms from 1 to 10 years. During 3 years the compensation of external and the tissue respiration was observed with further stabilization. Beginning from the fifth year in patients, to whom pulmonectomy and operation, causing complications, were performed, the signs of respiratory insufficiency appeared. Data obtained may be used for timely introduction of corrections into labor recommendations for prophylaxis of respiratory system activity decompensation in late period. PMID- 12549318 TI - [Reconstruction-restorative surgeries of mastitis sequelae]. AB - Experience of performance of reconstruction-restorational operations in 14 patients with consequences of postnatal mastitis (PM) was analyzed. In 2 observations the silicon endoprosthesis were applied, in 11--the musculocutaneous flap using broadest muscle of the back (BMB) and straight abdominal muscle, in 3- combination of both, BMB and endoprosthesis. Immediate and late results are good. Reconstruction of mammary gland using, constitutes the operation of choice because of near location of donor site, satisfactory blood supply and plasticity needed. PMID- 12549319 TI - [Surgical treatment of breast neoplasms in early stages]. AB - Analysis of literary data and own investigation results for the modern surgical methods of treatment of mammary gland cancer in early stages was performed. Indexes of patients survival after surgical removal of mammary gland (MG) and quadranthectomy did not differ. Preservation of MG constitutes great social and psycho-emotional significance for women-patients. PMID- 12549320 TI - [Diagnosis and results of treatment of congenital pyloric stenosis in children]. AB - Results of treatment of congenital pylorostenosis in children during last 15 years were studied. There was established, that degree of hypotrophy increases proportionally to duration of vomitive period. Changes of clinical signs of illness due to depression of immune defence, observation by divisional pediatrician with administration of spasmolytic preparation constitute the causes of late hospitalization to unit of pediatric surgery. Late results of treatment witness, that timely performance of operation constitutes an effective method. PMID- 12549321 TI - [Principles of antibacterial therapy of abdominal infections]. AB - The principles of antibacterial therapy of abdominal infection application were analyzed. Most frequent revealed exciters of abdominal infections, information about its sensibility to antibiotics, new approach to antibacterial therapy were presented. PMID- 12549322 TI - [A large strangulated paraesophageal hiatal hernia]. PMID- 12549323 TI - [Simultaneous surgical interventions in an elderly female patient]. PMID- 12549324 TI - [Diagnosis of intraperitoneal injury of the bladder]. PMID- 12549325 TI - [Rare observation of thyroid gland sarcoidosis]. PMID- 12549326 TI - [Site-directed mutagenesis of the dichloromethane dehalogenase gene from Methylophilus sp. strain DM11]. AB - In order to investigate the role of different residues of Methylophilus sp. strain DM11 dichloromethane dehalogenase for substrate binding, glutathione affinity, and catalytic activity, site-directed mutagenesis studies of the gene encoding the enzyme were carried out. The conserved tryptophane residue at 103 region was respectively substituted by phenylalanine, valine or asparagine. The conserved arginine residue at 109 region was substituted by leucine. The conserved tryptophane residue at 117 region was respectively substituted by tyrosine or phenylalanine. Six mutant enzymes were produced. Among them three possess lower activities, other three do not possess activity. The properties of the mutant enzyme W117Y are very different from wild-type enzyme. PMID- 12549327 TI - [The construction of Alcaligenes faecalis ntrC-lacZ fusion gene and its expression during association with rice roots]. AB - A broad host range vector pLA2917 containing ntrC gene or ntrC-lacZ fusion were constructed, namely pLAC1 and pLAC2. The plasmids pLAC1 and pLAC2 were introduced into A. faecalis wild type strain A1501 by conjugation, subsequently to abtain A15C1 and A15C2. The expression and regulation of ntrC gene of A. faecalis associated with rice roots was investigated under the condition of the associative nitrogen fixation using X-Gal decoration method, micrograph and ntrC partially deletion mutant. The blue precipitation was strongly existed in parenchyma cells as well as in the lateral root primordial. It showed that ntrC gene could express at much higher level in these sites. In the presence of ammonia, the number of multi-copy ntrC conjugatants colonized on surface of rice roots is higher than that of wild type A1501, and the colonization of ntrC mutant is weakest among these three strains. This provided an evidence that ntrC gene might be involved in procedure of colonization of A. faecalis to rice roots. PMID- 12549328 TI - [Chinese Chlorella viruses and their molecular biological properties]. AB - An extensive survey for Chlorella viruses revealed that Chlorella viruses are widely distributed over China. Eleven isolates of Chlorella viruses have been isolated, which lysis the Chlorella viruses sp. (strain NC64A). These isolates were named BJ-1, BJ-2, BJ-3, BJ-4, FJ-1, FJ-2, NJ-1, CDT-1, HCJ-1, SCB-1 and SCC 1 respectively. They have several common properties, including polyhedral morphology, and linear dsDNA genomes. However, the restriction patterns of viral DNA, the concentration of m5dC and m6dA in viral DNA, and the composition of viral structure protein are different among these isolates. All of viral major capsid proteins are 54,000 except FJ-1, whose major capsid protein is smaller than 54,000. Western blot analysis showed strong immunological cross reaction among all of viral proteins with PBCV-1's antiserum except FJ-1. It indicates that the homology between these isolates and PBCV-1 is high. Among these isolates, FJ-1 has some special properties. PMID- 12549329 TI - [Selection of drug-labelled Bacillus sphaericus and studies on transformation and expression of foreign DNA]. AB - NTG was used to make chemical mutation for Bacillus sphaericus, RifR and SmR labelled strains were selected, which could resist drug as much as 100 u/ml. The resistance to drug was stably inherited. The RifR strain was used as recipient and the plasmid containing the lysostaphin gene was transfered into it by protoplasts. Results showed that the lysostaphin gene could be expressed stably at high level in Bacillus sphaericus and the lysostaphin activity was about 122 u/ml medium after shaking culture. PMID- 12549330 TI - [The intergeneric compatibility of heredity and expression for cellulase genomes between Aspergillus niger and Trichoderma reesei]. AB - By using the developed display techniques of cellulase isozymes and the RAPD-PCR analysis guided by a deduced universal sequence of cellulase genes, the polymorphisms of genomic DNA fingerprints and cellulase isozymes were compared among three typical stable recombinants (3a, 3b, A7-1) and their two parents (Aspergillus niger AMS11, Trichoderma reesei QM9414) in order to provide the molecular evidence of gene recombination, to demonstrate the compatibility of heredity and expression of intergeneric genomes, and to assay on the molecular fundamentals of hybridization dominance. The results showed that in these recombinant strains the recombinantal fingerprints of genomic DNA could be stablly hereditary and the expression of recombinantal CMCase (carboxymethylcellulase) and beta GLase (beta-glucosidase) could be compatibly enhanced. The diversity of molecular fundamentals of cellulase hybridization dominance were (1) the compatible co-existence and enhanced expression of some hereditary beta GLase-coding genes from two parents in recombinant 3b; and (2) the compatibly enhancement of expression between the hereditary genes encoding beta GLase and CMCase from two parents, resulting in the dramatic increase of proteins of corresponding isozymes in recombinants 3a and A7-1. Based on these, a proposal model for the double synergism on cellulase activity in vitro and on its biosynthesis in vivo mediated by beta GLase was suggested. A practical method for assaying on the molecular fundamentals and the stability of hybridization dominance of recombinants was thereby established in this study. PMID- 12549331 TI - [Expression, purification and characterization of recombinant human cyclophilin A]. AB - Plasmid-derived expression of the human CyPA in E. coli would make it possible to obtain ample protein quantities and to avoid difficult task of obtaining human tissues for protein purification. The cDNA encoding human CyPA from MT4 lymph cell line has been cloned and an expression vector (pET11/CyPA) has been constructed under control of the T7 promoter for efficient expression in E. coli. The recombinant CyPA is produced at 41% of total soluble cell protein, and showed the peptidyl-prodyl cis-trans isomerase activity. PMID- 12549332 TI - [Sequence analysis of the serotype specific gene fragment of VP4 from three rotavirus field strains]. AB - Rotaviruses are the major pathogens that cause life threatening diarrhea in young children and animals. The 4th gene, encodes the nonglycosylated major neutralizing protein VP4 which can be cleaved into VP8 and VP5. The regions of the fourth genes coding for the VP8 protein, downstream cleavage site and the NH2 terminus of VP5 from three Beijing human rotavirus field strains (T65 [P1A], R50[P1B], N16[P2]) were cloned and sequenced. Comparative analysis of the deduced amino acid sequences of these 887 bp fragments from three strains indicates a high degree of homology (92%-96.6%) between field and reference strains within the same serotype and a high degree of divergence (70.5%-71%) among different serotypes. Notably, the most divergence of amino acid sequence is located in a region delimited by aa 84-172 of VP8 protein. In addition, the possibility of trypsin cleavage sites in virulence is discussed. PMID- 12549333 TI - [Studies on some properties of the immobilized penicillin acylase by polyacrylonitrile fibres]. AB - The extracellular penicillin acylase from Bacillus megaterium was immobilized by coupling to derivatives of polyacrylonitrile fibres. The apparent activity of the immobilized penicillin acylase was about 2,000 u/g (dry weight). The optimal pH and temperature were 9.0 and 50 degrees C for hydrolysis of penicillin G, respectively. The immobilized enzyme was stable in the pH range of 5.5-10.3, and at the temperature below 50 degrees C. The apparent Michaelis constant Ka and Vm of the immobilization enzyme were 1.33 x 10(-2) mol/L and 2.564 mmol.min-1, respectively. The inhibition constant of phenylacetic acid acted as a competitive inhibitor for The immobilization enzyme was 0.16 mol/L. The remained activity was about 80% after operating 20 times. PMID- 12549334 TI - [Studies on optimal conditions for inulinase production by Kluyveromyces sp. Y 85]. AB - The application of response surface method to optimal medium-screeing for inulinase production by Kluyveromyces sp. Y-85 has been investigated. The experiments of condition control for the enzyme production was carried out in a 15 L fermenter. A large scale fermentation (1000 L fermenter) was performed five batches, and average inulinase activity of 68.9 u/ml achieved. PMID- 12549335 TI - [luxAB genes as marker for detecting Rhizobium fredii HN01 nodulation functions]. AB - A suicide plasmid pHNC3 which contains Tn5-luxAB was transferred into Rhizobium fredii HN01 by the help of pRK2013. Then Tn5-luxAB inserted on the genome of HN01 and gave luminescence activity. The luminescence colonies were picked up and the Eckhardt gel was performed for plasmids profile detection. The location of Tn5 luxAB on the genome was determined using the luxAB as probe. The colonies which were marked by Tn5-luxAB on the chromosome and different plasmids of HN01 were chosen for pot experiment, and a chromosome labelled strain HN01LC02 was detected by soil pot experiment. The detections included the nodulation occupancy and the luminescent nodules distribution on the root system formed by the luxAB-marked rhizobia. PMID- 12549336 TI - [Studies on fermentation conditions of cytokinin produced by rhizobia strain 4012a]. AB - The conditions of cytokinin fermentation of the rhizobia strain 4012a were detected by the ELISA. The results indicated that the optimal medium for cytokinin production by strain 4012a was composed of glucose 10 g/L, (NH4)2SO4 1.0, K2HPO4.3H2O 0.6, MgSO4.7H2O 0.1, CaCl2.2H2O 0.4, FeCl3.6H2O 0.04, Na2MoO4.2H2O 0.1 mg/L, calcium pantothenate 100 micrograms/L, adenine 200 mg/L. When strain 4012a was grown in 250 ml flask containing 50 ml of the medium on the rotary shaker (150 r/min) at 27 degrees C for 96 h, the yield of CTK 908 micrograms/L culture solution was obtained. It displayed bioactivity kinetin equivalents (KE) 1 mg/L medium with the radish cotyledon expansion test. PMID- 12549337 TI - [Cloning and analysis of the regulatory gene fragment induced with seed extract in Rhizobium huakuii strain 7653R]. AB - The large plasmids of strain 7653R were digested with restriction enzyme EcoRI. Their DNA fragments were cloned into the expression vector pMP220 to construct a lacZ fusion pool, which were transferred into the recipient strain 7653R. Tri transconjugants were selected onto plates containing X-gal and seed extract. Five blue colonies were assayed of their beta-galactosidase activity after incubation with or without seed extract. A positive induced strain HN18 was obtained. Hybridization of nodDABC probe on the recombinant plasmid pHN18 showed a 1.7 kb positive band. The evidence makes a deduction that the pHN18 contains a promoter of nod operon. PMID- 12549338 TI - [Purification and properties of intercellular inorganic pyrophosphatase from Saccharomyces cerevisiae]. AB - An inorganic pyrophosphatase (EC3.6.1.1) from Saccharomyces cerevisiae was purified to PAGE homogeneity by sonication disruption, (NH4)2SO4 fractionation and DEAE-cellulose column chromatography. The optimum pH and temperature of the enzyme were 7.4-7.8 and 60 degrees C, respectively. The Km was 19.3 mmol/L. The enzyme required Mg2+ as a cofactor for hydrolysis of pyrophosphate and was inhibited by Ca2+, Hg2+, Pb2+, Mn2+. PMID- 12549339 TI - [Identificating of Actinomycetes producting activity of herbicide]. AB - 14 strains of Actinomycetes were isolated from soil samples collected in Yunnan, China. Experiments showed that they had activity of herbicide. In this paper, the morphological and chemotaxonomic characters of these strains were studied and they were identified as three genera of Actinomycetales respecitively, such as Streptomyces, Micromonospora and Nocardia. PMID- 12549340 TI - [Studies on the fermentation technology of microbiological preparation No.633]. AB - This paper reports the fermentation techmology of bacterial feed No.633. A new fermentational technic, which was composed of fermentation medium, optimum pH, temperature, airate flow, pot pressure and fermentation time, was established. PMID- 12549341 TI - [Bacterial phylogeny]. PMID- 12549342 TI - [A biomechanical evaluation of the effects of XW630 on the healing of experimental fracture]. AB - The aim of this study was to evaluate whether tetracycline-estrone (XW630) plays a role in the healing of fracture. 120 rabbits were divided into 4 groups and subjected to osteotomies at the middle of both radials, and they were given XW630, estrone (E2), tetracycline (T) and 0.9% sodium chloride (C) respectively after operation by means of intramuscular injection (M) or local injection(L) in the fracture region. The biomechanical properties, including bending and compression, and the biochemical changes were tested at 30 days, 60 days and 90 days after osteotomy. The results showed that the bending stress and compression stress in the XW630(M) group were significantly higher than those in other groups at different periods after osteotomy. At 30 days after osteotomy, the bending ultimate stress in the XW630(M) group was 80% higher than that in group C, 76% higher than that in group E2, and 69% higher than that in group T. At 90 days after osteotomy, the bending ultimate stress in XW630(M) group increased to 80% of that of the normal radius, and it was 30% higher than that of group C, 34% higher than that of group E2 and 19.5% higher than that of group T. The biochemical changes corresponded with the changes biomechanical properties. These data suggest that XW630 given by intramuscular injection plays an obvious role in accelerating the healing of fracture. The medicine given by intramuscular injection is obviously more effective than that locally injected. PMID- 12549343 TI - [Transient response in electric measurement of human auricular point]. AB - This is an experiment on the measuring E-t and R-t for human auricular point, which shows there is a transient response of electric potential E(t) and voltage drop U(t) when t < 2 tau and there is at intervals change with the time after 2 tau about R(t) and E(t). We have obtained a response function of the E(t) and U(t) by the circuit analysis. The parameter of the transient character is relaxation time tau,tau approximately RC. The mathematical model is in correspondence with the experimental result of the auricular point and TPM simulation. It indicates that the character changing with the time should be extracted after 2 tau. This experiment is of importance to understanding the electric character of the otopoint. PMID- 12549344 TI - [A method for lung zone recognition in X-ray film]. AB - Based on the lung anatomy, the border of lung zone is expressed as a set of parabolic and straight lines. After several feature points are detected, the parameters of the lines are determined and the lung zone in X-ray film is recognized. A coarse-to-fine strategy is employed in the procedure to lower the computation consuming. The method has been successfully used in an application system. PMID- 12549345 TI - [Simulation study of selectively stimulating nerve fibers using biphasic pulses]. AB - Biphasic selective stimulation is an effective electrical stimulation mode for preventing nerve fibers against electrochemical damage, for muscle to contract smoothly, and decreasing muscle's fatigue. By use of computer simulation, this paper presents three biphasic stimulation modes respectively using monopolr, bipolar and tripolar electrodes, which can effectively stimulate nerve fibers in a selective way. The results are helpful to the clinical application of neuroelectrical stimulation. PMID- 12549346 TI - [Effects of biomaterials on immunoglobulins and biochemistry in serum of extracoporal circulation]. AB - The polyvinyl chloride(PVC) tube coated with cellulose acetate(CA), polyether suffone(PES), polyurethane(PU), silicon rubber(SR) and polyester fibre(PETF) were put in contact with circular serum at 37 degrees C. According to different time periods of exposure, the immunoglobulins (G.A.M), albumin(ALB), total protein (TP) and total cholesteral(Tch) in serum were measured by an IL-Monarch biomedical analyser. The results showed that the levels of immunoglobutins significantly decreased after the serum came in contact with CA, PETF, SR and that the levels of ALB, TP and Tch were significantly lower in the serum exposed to PETF while those in the serum exposed to other materials were normal. These changes may relate to the complement activation induced by different materials. PMID- 12549347 TI - [Study on bone-repairing biomaterial ytterbium oxide-hydroxyapatite]. AB - Hydroxyapatite was extensively used to repair osteoshortage recently. To improve its bondibility and X-ray obstructibility, we added ytterbium oxide of different proportions in the synthetic process of hydroxyapatite. Ytterbium oxide is characterized by high activity and X-ray obstructibility. A cheap biomaterial with good biocompatibility and high X-ray obstructibility has been got in this way. PMID- 12549348 TI - [Experimental study on the biocompatibility and toxicology of interface between hydroxyapatite coating and bone]. AB - According to "The standards for biological evaluation of bio-materials and bio products abbreviated to STANDARDS" newly set by the Ministry of Health, PRC, 5 observation indexes were selected to evaluate systematically the biocompatibility and toxicology of China-made hydroxyapatite, which is clinicall used. The results revealed no effects of allergy, mutation and hemoclasis reaction except mild cytotoxicity. These indicate that the China-made hydroxyapatite meets the demand of STANDADS and thus may be clinically with safety. PMID- 12549349 TI - [Study on the method of combining albumin with biomaterials for selecting a substitute for hard ttissue of human body]. AB - This study explored the conditions on combining albumin of surfaces of hydroxyapatite (HA), bioglass ceramics(BGC) and hydroxyl poly-calcium sodium phosphate (HP) by using the method of BrCN activation. The result demonstrates that albumin can be combined with HP, but it can not be combined with HA and BGC. PMID- 12549350 TI - [The test of mean flow velocity patterns within a new push-plate ventricular assist device]. AB - This article reports the design of Z-4 Push-Plate Ventricular Assist Device and the test of its mean flow velocity patterns. The results showed that during diastole the interaction of the jet formed by incoming fluid within the boundaries of the blood chamber induced a large clockwise vortex, its centre was the centre of the chamber circle; the largest velocity (approximately 125 cm/sec) was generated near the major orifice of the valve in inlet conduit. During systole the centre of the vortex came close to the outlet conduit; the largest velocity (approximately 120 cm/sec) was generated in inlet conduit. There were no turbulent flow and stagnation point during the pume cycle. The mean velocity patterns proved a good wall washing over the entire chamber and the internal surfaces of the pump should be resistant to thrombus deposition. PMID- 12549351 TI - [Study on immobilized cells for producing alpha-amylase by using polyving alcohol as the carrier(II): The effect of fermentating conditions on the ability producing alpha-amylase of the cells immobilized with polyving alcohol as the corrier and continuous fermentation of the immobilized cells in CSTR]. AB - The effects of fermentating conditions on the ability of immobilized cells with PVA as carrier for producing alpha-amylase were studied. The continuous fermentation with the immobilized cells were tested in continuous flow stirred tank reactor (CSTR). The results showed that the adaptability of the immobilized Bacillus substilis to pH increased after immobilization. In CSTR, the immobilized cells can be fermentated continuously for 360 hrs and the activity of alpha amylase can be kept on the level of about 170 u/ml. PMID- 12549352 TI - [The time-frequency analysis of the heart sound]. AB - The Wigner-Ville distribution, the Choi-Williams distribution, and cone-kernel distribution are three time-frequency distributions. We compared the interference attenuation and time-frequency concentration properties of the three TFDs. The properties were then further investigated by studying the results obtained for a chirp signal. We analysed the heart sounds, nonstationary signals, with the three TFDs. The results indicate that the time-frequency signal analysis methods have high time-frequency resolution. The time-frequency signal analysis techniques may be helpful in unveiling the complex mechanisms of heart sound generation and be used in medical research and clinical diagnosis. PMID- 12549353 TI - [The changes of erythrocyte deformability and cardial function in coronary heart disease with various degrees of coronary stenosis]. AB - According to the extent of coronary maximal stenosis, 29 cases of coronary heart disease (CHD) were classified into two groups: (I) obvious stenosis (OS, narrowing > or = 50% in luminal diameter, n = 15); (II) mild stenosis (MS, marrowing < 50% in luminal diameter, n = 14). The results showed that erythrocyte deformability index (DI), left ventricular ejection fraction (EF%) and fractional shortening (FS%) in the CHD patients were decreased as compared with those in the control group (P < 0.05 or 0.01), and the DI, EF and FS in the OS group were significantly lower than those in the MS group (P < 0.05). These suggest that DI, EF and FS may be of predictable value in identifying CHD patients with or without significant coronary stenosis. PMID- 12549354 TI - [Hemorheologic change after strong blood flow disturbance caused by gunshot and its influence on the prognosis of the wounded animals]. AB - Three types of experimental models were used in this study: 1. Both thighs of dogs were wounded by 5.56 mm bullet. 2. Femoral artery and vein were tied at the time of wounding. 3. Shock caused by simply bleeding the dogs' femoral arteries. We observed the characteristics of change hemorheology and analyzed its effects on the wounded dogs. The results showed that strong blood flow disturbances always brought about hemorheologic changes such as rapidly increased blood viscosity and red cell's aggregation index and the simultaneous phenomenon of blood concentration. These may be related to extensive endothelium injury and destruction of a number of blood cells caused by disturbance wave. The above change may give rise to abnormality of microcirculation and disturbance of matter transmission; in that sense, it may provide the physiopathologic basis for the occurrence and development of the complication. PMID- 12549355 TI - [Finite element analysis of human tibia in vitro]. AB - This investigation was aimed at the fatigue fracture of tibia that often occurs in soldiers, as well as in dancers and athletes. The data of ground reaction force(GRF) of soldiers were measured using a force platform in three common gaits: marching, parading and running. The stress distribution in tibia under these GRFs were analyzed with a FE model. The results showed that the smallest GRF(744.8N) is in quick marching, the next (1156.4N) in parading and the largest (1391.6N) in running. The stress distribution under GRF was mot uniform in the tibia. The maximum stresses occurred at a site 1/3 tibia length below the knee where tibia fracture is often seen clinically. The maximum stresses were 110 MPa and 115 MPa in parading and running respectively. This means that the tibial tractures in soldiers are mostly associated with stress, i.e. the greater the stress is, the higher the incidence of stress fracture comes about. PMID- 12549356 TI - [A method for standardizing brain CT image]. AB - In this paper, we propose the concept of standardization of brain CT image and introduce a kind of standardization method based on geometry correction technique. To work out the method, first we preprocessed the raw CT image, second automatically searched the control points which were used for establishing the correction equation, and last we completed the standardization of the CT image. The result of the experiments demonstrated the pracficability efficiency of this kind of method which conld provide theoretical foundation and experimental means for the diagnosis of brain disease. PMID- 12549357 TI - [Collagen sponge and its hemostatic properties]. AB - Soluble collagen was extracted from bovine tendon in acid solution. Collagen sponge hemostatics were prepared by means of lyophibization. The amino acid and UV spectrum analyses were made to confirm the composition of the soluble collagen. The results of experiments on rabbits indicated the collagen sponge prepared in the authors' lab had excellent hemostatic and adhesive properties in vivo surgical tests. PMID- 12549358 TI - [Peripheral artery blood flow measurements using pulsed Doppler and electromagnetic flowmetry in dogs: a comparative study]. AB - To investigate the accuracy of blood flow parameters in peripheral artery by Doppler ultrasound, the intermal diameter(D), peak velocity(Vp), mean velocity (Vm) and flow volume (Q) of dog's femoral artery were measured by using either a pulsed Doppler (PD) system or electromagnetic flowmeters(EMFM). The results showed that the D measured by B-mode ultrasound values significantly smaller than that by callipers but it correlated very well with the D by callipers. When the values of Vp, Vm and Q measured by pulsed Doppler and EMFM were compared, the Vp had the most highly significant correlation between the data obtained by the two techniques (r: 0.91, 0.79, 0.83 respectively). When Vp, Vm and Q were measured by angle correction of PD, the correlations between the data determined by the two techniques were still significant(r: 0.88, 0.82, 0.78). PMID- 12549359 TI - [Study on antibiotic rate-limiting barrier membrane by radio-frequency glow discharge plasma deposition]. AB - A method that forming antibiotic rate-controling membrance on surface of polyurethane and ciprofloxacin matrix by RF-GDPD was studied. The surface characterization of film was analysed by XPS. The elution of antibiotic drug in vitro was discussed. PMID- 12549360 TI - [Research and application of cardiopulmonary-cerebral resuscitation]. AB - A novel concept of cardiopulmonary-cerebral resuscitation (CPCR) has been proposed, in which a combination of technologies of the membrane artificial lung, mild hypothermia, centrifugal blood pump, heparin coated surfaces and blood shunting have been used. The present paper introduces the modern conception, history of CPCR, with emphasis on the brain(cerebral) resuscitation. Importance of extracorporeal membrane oxygenation, mild hypothermia and centrifugal blood pump in cerebral resuscitation has been emphasized. Thereupon the new idea of the novel CPCR has been proposed and the design and laboratory manufacture of the equipment of CPCR will be carried out. PMID- 12549361 TI - [Absorbable self-reinforced biomaterials used for fracture fixation implants]. AB - A series of absorbable polymers used for osteosynthesis implants are introduced in this review. To reduce the degradation rate and increase the strength of them, some methods, such as self-reinforced or coating with other polymers, have been presented. Some clinical and animal's experiments have been discussed. PMID- 12549362 TI - [Phase aberration correction for ultrasound imaging]. PMID- 12549363 TI - [A method for measuring the isocentre positional accuracy of X knife system]. AB - The mechanical isocentre positional accuracy of the X Knife system is a very important technological parameter in the course of the X Knife therpy. In this paper, a film-concentric circle method for measuring the mechanical isocentre positional accuracy is introduced. A simulation ball set at the isocentre of the X Knife mechanical positional system, as a substitute for target, was imaged on a film (the simulation ball's image is a circle and its diameter is 8 mm), and a collimator was imaged on the same film (the collimator's image is also a circle and its diameter is 30 mm). The distance between these two circles' centres is the error of the isocentric system. This method has been used in our work. PMID- 12549364 TI - [Check-up of the hyperbaric chamber and attached equipment]. AB - This article is about the check-up of the constructed and repaired hyperbaric chamber and attached equipment. The auther proposes guidelines for checking hyperbaric chamber body and pressure container, AC 380 V electric circuit, AC or DC 220 V below electric circuit system and instrument, mechanic equipment and piping system, air-conditioner system, test of the hygiene index, the rules and regulations of chamber, etc. PMID- 12549365 TI - [Expression of the polyketide ketoreductase gene from midecamycin producing strain in E. coli]. AB - Two primers were designed and synthesized according to the polyketide ketoreductase from midecamycin producing strain gene(MKR) sequence data which has been reported, MKR gene was amplified using PCR technique. The amplified MKR gene was subcloned into NdeI and BamHI sites of the T7RNA polymerase-dependent pT7-7 expression vector, and introduced into E. coli K38/pGP1-2. Proteins were isolated from transformant. The result of exclusive labeling by L-35S-methionine of plasmid-encoded proteins SDS-PAGE analysis showed that the MKR was produced in E. coli. The expressed MKR has bioactivity. PMID- 12549366 TI - [The construction of poliovirus chimera and the analysis of its antigenic structure]. AB - The construction of poliovirus chimera is an useful way to investigate the antigenic structure conformation of poliovirus. In this research, the poliovirus chimera containing a piece of antigenic fragment of hepatitis A virus was constructed for analysis of poliovirus antigenic structure conformation. Depending on the characterization of this chimera, the conformation of poliovirus neutralization antigenic site I in VP1 was analyzed, and that the possible influence of hepatitis A virus antigenic fragment inserted to the structure conformation of poliovirus antigen was also investigated. PMID- 12549367 TI - [Comparative research on two different killer strains of Saccharomyces cerevisiae]. AB - Using two different killer strains SK4(K1 type) and ERR1(K2 type), their killer characters were analysed and find two strains compete with each other when they are mixed and killer characters are related with the growth of strains. Different M-dsRNA plasmids in different killer strains have different molecular lengths, being 1.7 kb and 1.5 kb in SK4 and ERR1, respectively. L-dsRNA plasmids in both strains have the same molecular lengths, 4.0 kb. Cured by high temperature (38 degrees C) and U.V. (15 W, 30 cm, 5 min), killer strains lose their killer characters with the disappearance of M-dsRNA, the curing rate of killer activity is associated with strains and curing conditions. Killer toxins produced by two strains have different killer effects. Toxins of SK4 and ERR1 have the different highest killer activity at pH 4.8, 16 degrees C and pH4.2, 22 degrees C, but both of two toxins have the greatest effect on the sensitive strain in the late-log phase. PMID- 12549368 TI - [Purification and characterization of cyclomaltodextrin glucanotransferase from alkaliphilic bacteria]. AB - A cyclomaltodextrin glucanotransferase from alkaliphilic bacteria was purified to PAGE homogenous by ammonium sulfate precipitation and DEAE-cellulose DE-52 chromatography with 11.5 fold purification and 5.7% recovery. Mr estimated with concentration gradient PAGE was 151,000. The optimum conditions for activity were pH 7.0, temperature 60 degrees C, stable in the range 6.0-9.0 and below 50 degrees C. The enzyme activity was activated by salicin and maltitol, slightly inhibited by Sn2+, Mn2+, inositol and pullulan, strongly inhibited by Ag+, Cu2+, Al3+, Fe2+, Pb2+, Hg2+ and Zn2+. The maximum absorption was at 270 nm in ultraviolet spectrum, the maximum emission wavelenghth in the excitation spectrum was 283 nm, the maximum emission spectrum determined at 283 nm was 335 nm. The effects of some protein modification reagents on the CGTase activity have been studied. Histidine and tryptophan residues may be essential for activity, carboxyl groups might have some effects on the activity. PMID- 12549369 TI - [Studies on the selection of strains producing colominic acid and culture conditions]. AB - One strain E. coli C-8, the highest yield of colominic acid, was selected from 40 E. coli strains in the medium in which glucose and ammonium sulfate were the only carbon and nitrogen resources. An optimum medium for the growth and colominic acid production of E. coli C-8 was studied. The optimum carbon resources for colominic acid production was sorbitol selected from 16 kinds of carbons, and its optimum concentration was 2.5%. The optimum inorganic and organic nitrogen resources for colominic acid production were ammonium sulfate (0.5%) and tryptone (1.5%), respectively. The optimum concentration of K2 HPO4 was 90 mmol/L. The optimum culture temperature was 37 degrees C, but no colominic acid was produced below 20 degrees C. The optimum pH range was 7.5-8.2. The strain growth in the optimum medium kept logarithmic phase in 40 h (maxim A = 22). The colominic acid secreted into medium was much lower before 20 h, but high biosynthetic rate of colominic acid was detected after 40 h, the colominic acid reached the highest level (1,200 micrograms/ml) at 65 h. PMID- 12549370 TI - [Studies on reduction of hexavalent chromium by fungi]. AB - Several fungal strain, isolated from activated sludge, sewage and soil, were able to grow in the medium containing 300-500 mg/L K2Cr2O7, and strain BS-1 was capable of resistance to K2Cr2O7 up to 900 mg/L. After 4-6 d growing of the strain BS-1 and other 3 strains in the liquid medium added 200 mg/L K2Cr2O7, the Cr(VI) content of the medium disappeared completely. These strain were identified as Penicillium sp. BS-1, BS-3, Aspergillus niger BR-4, Aspergillus flavus BX-1 respectively. The results from the detection of the fungal culture by UV absorbance spectrum and chemical analysis showed that high-toxic Cr(VI) was reduced to lesstoxic Cr(III). Reaction conditions and effectors of Cr(VI) reduction by intact cells of Penicillium sp. BS-1 were studied. The optimum temperature was 30 degrees C, the optimum pH was 7.0. Addition of glucose (0.25%) enhanced Cr(VI) reduction, but high concentrations of Cr(VI) inhibited the reduction of Cr(VI) to Cr(III). PMID- 12549371 TI - [Saccharification of corn stover by immobilized Trichoderma reesei cells]. AB - The mycelia of Trichoderma reesei Rut C30 were adsorbed and immobilized within the porous polyurethane supporter. It was found that the synthesis of cellulase by the immobilized cells and the enzymatic hydrolysis of corn stover were able to be carried out in a nitrogen source-limiting medium. Under repeated batch processes with 60 g/L corn stover pretreated by 2% NaOH at 85 degrees C, the average cellulase activity (FPA) was 0.70 IU/ml, the concentration and yield of the reducing sugar were 26.41 g/L and 89.11% respectively after shaking culture at 150 r/min, 30 degrees C, pH 4.8 for 108 h. Total of 12 repeated batches were performed in 54 days. The immobilized cells kept the weight around 10 g/L. The fed-batch process was also studied with the same immobilized cells. Total of 120 g/L corn stover was added in different feeding manners. The reducing sugar of 52.8 g/L was produced after 7 days and the saccharification efficiency (89.2%) was almost the same as the batch process. The results indicated that the cellulase production and cellulosic material saccharification in situs by the immobilized Trichoderma reesei cells is an convenient and effective process for conversion and utilization of renewable biomass. PMID- 12549373 TI - [A study on microbiological characteristics of upflow anaerobic filter]. AB - This experiment was conducted to study the relationship between performance and bacterial population in upflow anaerobic filter by improved Hungate technique. The results obtained showed as follows: (1) There existed correlation closely between the bacterial population, COD and volatile fatty acids removal; (2) The bacterial population was affected to a small extent by the change of organic loading rate in short time and suspending performance; (3) Successful performance of the reactor depended on the microbes' composition and their coordinated metabolism. PMID- 12549372 TI - [Purification and properties of inulinases from Aspergillus niger M89]. AB - Four inulinase components were purified from Aspergillus niger M89 by (NH4)2SO4 fractionation, Sephadex G-200 gel filtration, DEAE-cellulose chromatography and polyacrylamide gel electrophoresis (PAGE). The molecular weight of EI EII EIII and EIV were indicated to be 102.6, 97.9, 62.5 and 36.5 kD respectively by SDS PAGE. Measured by isoeleetric focusing, their isoelectric points were 4.15, 4.24, 4.48 and 4.15 respectively. Four forms of inulinase exibited maximal inulinase activity between 55-60 degrees C with optimal pH 4.0-5.0. There were some difference in their thermal stability. The little the molecule of the component was, the stronger its thermal stability was. All of four components were exo acting inulinases. PMID- 12549374 TI - [Studies on microbiological factor in colour change of mogao graffito's mural. II. Effect of microorganism on the pigment of imitative mural]. AB - Throung the assay and analysis of the imitative mural after cultivated with microorganisms, it has been shown that microorganisms had a pronounced effect on the pigments of the mural: the pigments secreted by microorganisms changed the colour of the mural, and produced much oxalic acid salt, which damaged the formation of crystals of the pigment, moreover, the chemical combination valence of Pb3O4 had been changed due to the metabolic products of microorganisms, that may play an important role in the chemical change of Pb3O4. PMID- 12549375 TI - [Studies on actinomycetes population from salt lake of Yuncheng in Shanxi]. AB - 120 strain of mesophilic and thermophilic of alkalaphilic actinomycetes of alkalic tolerant actinomycetes isolated from Yuncheng salt lake in Shanxi were identified. On the basis of morphological characteristics and some chemotaxonomic properties (composition of cell wall, whole cell sugar), the isolated strains were classified into Nocadiopsis, Micromonospora, Actinomadura and Streptomyces. The Streptomyces strains were classified into 11 groups. PMID- 12549376 TI - [Sequencing of a beta-amylase gene from Bacillus firmus]. AB - The gene encoding a beta-amylase from Bacillus firmus 725 was sequenced. The sequenced DNA of 2012 bp contains one open reading frame of 1406 nucleotides without a translation stop codon. The deduced amino acid sequence homology with those known bacterial and some plant beta-amylase was 98% for Bacillus polymyxa 72, 98% for Bacillus polymyxa ATCC8523, 82% for Bacillus circulans, 54% for Clostridium thermosulfurogenes, 49% for Bacillus cereus BQ10-S1, 50% for Bacillus cereus var. mycoides, 36% for barley, and 36% for soybean Eleven well-conserved regions were found among the amino acid sequences of the nine beta-amylases. PMID- 12549377 TI - [A new genus of oral bacteria in human]. AB - A strain, No. 90-1, is isolated from the oral cavity of a patient with periodontophthy. This strain is a Gram-positive, non-endospore-forming, facultative anaerobe with spherical cells, 0.9-1.5 microns in diameter, occuring in pairs and seldom in short chains of four cells, and motile by one flagellum per cell. The optimum growing temperature is 35-37 degrees C; appreciable growth is not found below 10 degrees C, but growth at 53 degrees and tolerance to 60 degrees C for 30 min. This strain is microhalophilous and grows best, well and poorly in the medium containing 2%, 10%-15% and 25% NaCl respectively. Catalase and urease are positive and nitrate is reduced. Acid is produced from many carbohydrates, but no gas. Gelatin can be hydrolyzed, but starch, cellulose and dextrin do not. G + C content in DNA is 41.34 mol%(Tm). The strain(90-1) is considered to be a new species belonging to a new genus because its some characteristics are different from those of the known coccus genuera and designated as Stomatostreptococcus microhalophilus Ping, Zhou, Sun et Fan gen. nov. sp. nov. according to its source and microhalophilic trait. PMID- 12549378 TI - [An investigation on multiple drug resistance of the rifampin resistant strains of Mycobacterium tuberculosis]. AB - The correlation between rifampin resistance and multiple drug resistance in 236 clinical isolates of Mycobacterium tuberculosis was investigated in this thesis. It has found that 99.4% of the strains with rifampin resistance were multidrug resistant strains and 89% of the multidrug-resistant strains were resistant to rifampin. This result showed that the rifampin resistance of Tuberculosis baccilli could be used as the marker of multidrug resistance of Mycobacterium tuberculosis. PMID- 12549380 TI - [Application of PFGE in fungal genetics]. PMID- 12549379 TI - [The experimental study on competitive PCR for quantitation of herpesviruses]. AB - A single pair of oligonucleotide primer selected within a highly conserved region of the DNA polymerase gene in herpesviruses was synthesized. The competitive template DNA purified from cytomegalovirus (CMV) DNA was used to carry out competitive PCR amplification with herpes simplex virus type 1 (HSV1) DNA (target sequences). And anti-HSV1 effects of acyclovir (ACV) was investigated by the method. The results showed that the efficacy of PCR amplification was equal to each other(the ratio of the quantity of competitor template with DNA to the target sequence was 1.5:1). As the concentration of ACV was increased, the quantity of HSV1, DNA was decreased. It suggests that this method is practicable and some defects of mutant template can be overcomed. PMID- 12549381 TI - [Expression characteristic of Bacillus thuringiensis cry1 gene in Pseudomonas fluorescens Pfx-18]. AB - The plasmids of lepidopteran-specific Bacillus thuringiensis strain from our laboratory were hybridized with RNA probe of cry 1Aa EcoR I-F fragment labelled using DIG. Cry1 gene was located in 39.3 MD plasmid. The plasmid was digested with Hind III and analysed by southern blot. It appeared both 7.1 kb and of 6.5 kb positive bands. The 7.1 kb fragment was ligated to broad-host-range vector pSUP106 and transformed into Pseudomonas fluorescens Pfx-18. The cloned strain, LZP-1 was obtained. The plasmids of LZP-1 were analysed by PCR. The results showed that gene-type is cry1Ab. SDS-PAGE analysis demonstrated that LZP-1 could express 66 kD insecticidal crystal protein and some small molecular weight peptides. Bioassay showed that motality of 1000-fold diluted fermentation broth was 33% to 3rd instar plulella xyloslelly larvae. PMID- 12549382 TI - [Identification of a sulfhydryl-reducing agent-inducible protein highly homologous to protein synthesis elongation factor Tu in Bacillus subtilis]. AB - It was examined that the effect of beta-mercaptoethanol and dithiothreitol treatments, which should affect disulfide bond formation of proteins, on cellular protein components of Bacillus subtilis. In LB medium, the treatments induced the synthesis of a 50 kD protein (P50), which is synthesized constitutively under normal growth condition and is a major cytoplasmic protein. P50 was also induced by heat shock, but not by sporulation. In Schaeffer's sporulation medium, however, P50 was not induced by the sulfhydryl-reducing agents. This suggests that the sulfhydryl-reducing agent-inducibility of P50 might depent on specific physiological condition(s). The amino terminal sequences of two of the four main V8 protease fragments of P50 were determined. A search in databases revealed that P50 was highly homologous to protein synthesis elongation factor Tu of B. subtilis. PMID- 12549383 TI - [Purification and properties of antifungal protein X98III from Bacillus subtilis]. AB - Bacillus subtilis BS-98 is one of the antagonistic strains strongly against plant fungal pathogens such as Physalospora piricola Nose. The antifungal protein was purified by ammonium sulphate precipitation and column chromatography on Sephadex G-100 and DEAE-cellulose, and it was named X98III. Molecular weight of X98III is 59,000 by SDS-PAGE and PI value is 4.50 by PAG-IEF, respectively. X98III was demonstrated as glycoprotein and lipoprotein by CAM (cellulose acetate membrane) electrophoresis and special staining. We estimated it contains 6% saccharides by using DNS methods. This protein was also found to be thermostabale and partially sensitive to proteinases. The amino acid analysis of the protein X98III showed that it comprises of 11 different amino acids and Glu, Tyr, Cys are the abundant amino acids. No Asp, Phe and Met were found. Purified X98III has strong inhibiting activity against the pathogens of Physalospora piricola, Phoma asparagi, etc. The antifungal mechanism of X98III was mainly disintegration of the cell wall to make the hyphae abnormal and the spores germinate abnormally or can not germinate at all. PMID- 12549384 TI - [A comparison of amino acid composition of proteins from thermophiles and mesophiles]. AB - The thermophilic feature of thermophiles are due to the high thermostability of their proteins. In order to investigate the mechanism of the high thermostability of thermophilic proteins and compare the difference of amino acid composition between thermophilic and mesophilic proteins, 110 pairs of homologous protein sequences from thermophiles and mesophiles respectively are collected and the amino acid composition, percentage of hydrophobic amino acids, hydropathy index, percentage of charged amino acids of the two protein groups are compared. It reveals that the two groups have evidently different composition in several kinds of amino acids, and thermophilic proteins have higher average hydropathy and charged amino acids composition. A comparison of the aliphatic indices between the two protein groups indicates that the reason that thermophilic proteins have higher aliphatic index is that thermophilic proteins have higher leucine composition, and thus sheds doubt on the validity of aliphatic index. It shows that it is possible to disclose some general rules with regard to the mechanism of protein thermostability by comparing a large amount of protein sequence data. PMID- 12549385 TI - [Research on the regulation of glucoamylase gene(glaA) expression in A. niger. II. Analysis of the function of 5'-regulatory region of A. niger T21 and 3.795 glaA gene]. AB - Two plasmid vectors pXH2 and pGH1 were constructed through the fusion of E. coli hph gene, the report gene and the 5' upstream regions of A. niger T21 and 3.795 respectively, as well as the terminator of A. nidulans trpC gene. The plasmid vectors were than used to transform A. niger T21 to functionally identify those different basic groups between the two 5' upstream regions responsible for high level expression of the glaA gene. Southern analysis of two transformants XH2C and GH1C revealed that pXH2 and pGH1 were integrated respectively into the chromosome at same site with two copies in tandem array. The level resistant to HmB(3000 micrograms/ml) of XH2C was twice as high as that (1500 micrograms/ml) of GH1C, indicating that the changes of basic groups through mutation result in twice increase of functional level of region responsible for transcription and regulation of A. niger T21 glaA gene compared with that of 3.795. PMID- 12549386 TI - [Nucleotide sequence analysis of an antibiotic biosynthesis gene of Streptomyces globisporus]. AB - Antitumor antibiotic C-1027 produced by Streptomyces globisporus has very high biological activity both in vivo and in vitro. Research works showed that one of biosynthesis gene of C-1027 is in the F2 DNA fragment. The plasmid pUC18 was used as vector to subclone the F2 DNA fragment. The nucleotide sequence analysis for F2 DNA fragment was carried out. Results showed that there is an open reading frame encoding for 122 amino acids. According to EMBO and GeneBanks data, this sequence may be a new one. PMID- 12549387 TI - [Purification and properties of alanine dehydrogenase from Streptomyces lincolnensis]. AB - Alanine Dehydrogenase (L-Alanine: NAD+ oxidoreductase, deaminating, EC 1.4.1.1) was purified from Streptomyces lincolnensis through four steps: (NH4)2SO4 precipitation, DEAE-cellulose 52, Affi-Gel Blue and Sepharose 6B. Molecular weight of the enzyme was determined as 170,000 by gel filtration and concentration gradient PAGE. SDS-PAGE showed only one band of 42,500, demonstrating that ADH from Streptomyces lincolnensis was consisted of four identical subunits. The optimal pH for amination was 9.0, for deamination 9.5. The optimal temperature for both amination and deamination was 50 degrees C. The Km valuse for pyruvate, NH4+, NADH, L-Ala and NAD+ were 2.08 x 10(-4) mol/L, 2.00 x 10(-2) mol/L, 2.38 x 10(-5) mol/L, 1.43 x 10(-2) mol/L and 6.67 x 10(-5) mol/L, respectively. PMID- 12549388 TI - [Degradation of phytic acid in rapeseed meal by two strains of molds]. AB - By using selective media, two strains of molds which are able to degrade phytic acid were isolated from soil samples. These molds are able to use inositol as the sole carbon and energy sources for growth. In broth cultures, the degradation rates of phytic acid by these molds were 74.4% and 95.0%, respectively. However, in solid-state fermentation which used rapeseed meal as a raw material, the degradation rate was about 40%. Several metal ions tested were able to enhance the degradation rate of phytic acid. Factors such as temperature, pH and moisture content which affect the growth of molds were also explored. These two strains of molds were initially identified as Paecilomyces sp. and Penicillium sp.. They do not produce any detectable amount of aflatoxins. PMID- 12549389 TI - [Studies on microbial factor on color change of Dunhuang mural. I. Classification of microbes on color-changed mural and property of some typical species]. AB - Microbial strains were isolated from 51 typical color changed samples of mural in 6 grottos of Dunhuang Mogao Gretto. After identification, it is proved that belong to 6 genera of bacteria and 5 genera of mould. Bacillus, Alcaligenes and Penicillium were dominant species in all of them. In the imitative experiment, it is found that Cladosporium sp., A. niger and two strains of bacteria have a great effect on the color change of red pigment of mural and aging of sizing agent. PMID- 12549390 TI - [Isolation and characterization of a insecticidal protein from Pseudomonas pseudoalcaligenes]. AB - A insecticidal protein was purified by gel-filtration chromatography on Sephadex G-100 and anion-exchange chromatography on DEAE-Sephadex A-50 from the suspension of Pseudomonas pseudoalcaligenes culture. Certain biophysical and biochemical properties were also studied. The molecular weight of the subunit of the insecticidal protein is 25,100 and pI is 5.16. Amino acid composition analysis showed that it is an acidic protein. PMID- 12549391 TI - [The biodegradation of trichloroethylene by a methanotrophic bacterium]. AB - A Methylomonas (strain GYJ3) isolated in our laboratory was identified as the type II methanotroph on the basis of the intractytoplasmic membrane of the ultrastructure. The optimal culture conditions for production of the soluble from of methane monooxygenase (MMO) were determined, in which the ratio of methane to air in atmosphere was 2 to 1 and Cu2+ concentration was 1.5 mumol/L. The biodegradation of trichoroethylene(TCE) by the resting cells of the strain GYJ3 was studied. All experiments were performed with cells grown under above conditions and thus expressing soluble MMO. This results showed that TCE at the high concentration of 30 mg/L did not inhibit to the enzymes in the cells. Addition of formate increased the initial specific TCE degradation rates. The product of TCE oxidation was found to be toxic to the cells. The degree of inactivation of MMO was proportional to the amount of TCE degraded. The TCE degradation capacities(Tc) of resting cells was determined. In no-formate and formate-fed experiments, the TCE degradation capacities were found to be 0.0778 and 0.0851 mg of TCE/mg of dry cell, respectively. PMID- 12549392 TI - [Strain of screening high toxin--producing from Corynebacterium diphtheria PW8 weissensee]. AB - Corynebacterium diphtheria PW8-weissensee was grown on different media for screening high toxin produce. Through six times of screening (selecting butyrous brilliant smooth small colonies and testing their toxin production capacity) from original strain producing toxin 160 Lf/ml, a high toxin--producing descendand culture was obtained, which produced 79% more toxin than the original strain did. PMID- 12549393 TI - [Study on the physiological activities of the metabolites from Paecilomyces arovirens]. AB - The metabolites of Paecilomyces arovirens exhibited insecticidal activities to many inscct pcsts, including cotton aphid, Aphis gossypii, hawthorn Ispidcr mitc, Tetranychus viennensis and larvae of imported cabbage worm, Pietis rapae. A comparison was made between the metabolites of P. arovirens and naphthyl acetic acid (NAA) in wheat (Triticum aestivum) and water melon (Cucumis sativus) bioassays on their physiological effects. Results demonstrated that some of the metabolites physiological activities of P. arovirens are same to that of NAA, some are different between them. It indicate that there is a new kind or many kinds of plant growth regulator in the metabolites and need further analysized. The results can offer reference for exploiting a new insecticid which not only can control insect pests, but also increase plant growth. PMID- 12549394 TI - [Coreceptors for HIV-1 infection]. PMID- 12549395 TI - [The envelope formation of AcMNPV and the location of envelope protein gp64 in host cell]. AB - Electron Microscopic study on the recombinant AcMNPV infected Sf9 cell showed that the virus has at least two forms of envelope formation: one is budding from plasma membrane and the other is enveloping in the nucleus by membrane structure. An additional possible pathway is budding from nuclear membrane. The envelope formation of AcMNPV in Sf9 cell line and its relationship with the envelope protein gp64 were investigated by using immunofluorescence techniques. The results showed gp64 was mainly located on the plasma membrane and nuclear membrane of infected cell. Whereas, there is no gp64 on the envelope of OV virus particles in the nuclei. This pattern cause the differences in envelope composition between the virus particles that budding from plasma membrane and nuclear membrane and those that enveloping in the nucleus. PMID- 12549396 TI - [Internalization and replication of Edwardsiella tarda in HEp-2 cells]. AB - It is demonstrated that Edwardsiella tarda possesses the ability to invade cultured epithelial HEp-2 cells by both lysis-counting assay and thin-section electron microscopy. Among fifteen strains of E. tarda, 6 strains internalized into HEp-2 cells and located mainly in vesicles. After internalization, the bacteria replicated in the host cells and then released into medium. Pretreatment of the HEp-2 cells with various concentration of cytochalasin D (0.1-5.0 micrograms/ml) or cytochalasin B (2.5-10.0 micrograms/ml) significantly reduced the amount of internalized bacteria in a dose-dependent manner. While pretreatment of HEp-2 cells with 0.25-100 mumol/L colchicine did not show any effect on the invasiveness of E. tarda. This strongly indicates that microfilaments are required for the internalization of E. tarda into HEp-2 cells and that microtubules are not involved in the entrance of E. tarda into HEp-2 cells. PMID- 12549397 TI - [Numerical taxonomy of bifidobacteria and homology analysis of some representative strains]. AB - Fifty five strains of bifidobacteria belonging to Bifidobacterium from various sources were studied by using numerical taxonomic techniques. These organisms included 13 reference strains and 42 newly isolated strains (19 strains of human sources, 25 strains of animal sources and 11 strains obtained from sewage). Several strains isolated from the animals weren't isolated before. Overall similarities of the strains for 75 characters contained morphological, physiological and biochemical characteristics and antibacterial susceptibility were determined. Dendrogram of numerical analysis based on unweighted pair cluster average linkage were achieved. Five clusters were defined at 70% Sm level. The relationships of these strains among five clusters were analysed. The strains of human sources were basically separated from the animal strains, and the sewage strains were distributed between the animal clusters and the human clusters. On the basis of the numerical analysis, the G + C content of DNA of some representative strains among these clusters were measured. Biotin-labeled DNA segments, complementary to variable regions of 16S rRNA, were amplified using PCR as probes. In the dot-blot hybridization the relationships of DNA segments among some strains were discussed. The test results showed that the homologies among the strains in same species and various species were different. The studies are valuable for further research in the relationships of some species among bifidobacteria of various sources. PMID- 12549398 TI - [Nucleotide and protein sequence analysis of rice dwarf virus replicase(segment S1)]. AB - The full length cDNA of Rice Dwarf Virus (RDV) Fujian isolate genome segment S1 was cloned and full length sequence was determined. The results showed that S1 is 4422 bp in length and contains a major open reading frame which encodes a polypeptide with 1444 amino acids. The major ORF contains the RNA-dependent RNA polymerase (RDRP) consensus sequences, such as motif I (DXXXXD) motif II (SGXXXTXXXN) and motif III(GDD). In addition to the three motifs, a well conserved region (EXXKXY) is found following motif III. Therefore, we suggest that the protein encoded by S1 encodes the virus RDRP. Homology comparison of the nucleotide and amino acid sequences of the RDV Fujian isolate with the known sequences of the Japanese isolate showed that the both isolate have 95% and 97% on nucleotide and amino acid sequences, respectively. PMID- 12549399 TI - [Construction and identification of recombinant fowlpox virus for expressing haemagglutinin-neuraminidase glycoprotein of Newcastle disease virus strain F48E8]. AB - For the construction of transfer vector pFGHN1175-1, the gene encoding haemagglutinin-neuraminidase(HN) glycoprotein of newcastle disease virus(NDV) strain F48E8 was removed from plasmid pGEMHN, and inserted into the HindIII site of insertion vector pFG1175-1, downstream of P7.5 promotor. Chicken embryo fibroblast (CEF) cell cultures which had been infected with fowlpox virus (FPV) Chinese vaccine strain 282E4 for 3-4 hours were transfected with pFGHN1175-1 plasmid DNA by liposomal transfection. Recombinant FPV with blue plaques were selected and purified 3 times in CEF cell cultures overlaid agarose containing X gal. PCR analysis and DNA dot-blotting hybridization assay indicated that the HN gene had inserted into the FPV genomic DNA. The expression of the NDV HN gene in recombinant FPV was confirmed by indirect immunofluorescence assay with specific monoclonal antibody. PMID- 12549400 TI - [Studies on microbial factor on color change of Dunhuang mural. III. Effect of Cladosporium sp. on color-change of red lead in the surface of mural]. AB - The conditions isolated Cladosporium sp. from Mural of Dunhuang grottoe germinated on the surface of imitated mural were as follows: 20 degrees C RH 60% or 30 degrees C RH 50%, which could be reached occasionally in some grottoes. Bone glue had protective function on red lead, while Cladosporium sp. could decompose bone glue and form some metabolite (such as oxalic acid, etc) as it grows. Then it could form a special chemical circumstance, which promoted the change from red lead to while lead. PMID- 12549401 TI - [Growth of Methylomonas Z201 cells and production of epoxypropane in monophasic and biphasic fermentation systems]. AB - The growth of Methylomonas Z201 cells and production of epoxypropane in mono- and biphasic fermentation systems were studied. In monophasic fermentation systems, the inhibitions of propene and epoxypropane on the growth of Methylomonas Z201 cells were observed and the concentration of epoxypropane reached 1.3 mmol/L. In biphasic fermentation systems, hexadecane acted as the "reservoir" of growth substrate (methane) and reactants (propene, molecular oxygen), the decrease of propene and epoxypropane in aqueous phase reduced the inhibition effect of propene and epoxypropane on the growth of cells, the concentrations of epoxypropane in both water phase and hexadecane phase reached 1.7 mmol/L and 2.6 mmol/L. In both monophasic and biphasic fermentation systems, the operational stability of cells was enhanced compared to that of resting cells. PMID- 12549402 TI - [Part of biological activity of peptidoglycan from Streptococcus lactis SB900]. AB - Peptidoglycan of Streptococcus lactis SB900(LABPG) was isolated. It's chemical composition was analyzed and part of biological activity was examined. The PG contained 9.84% protein, 0.871 mumol/m NAG, 1.14 mumol/mg NAM. The amino acids of relatively high concentrations were Ala, Glu, Asp and their concentrations were 1.046, 0.775, 0.304 mumol/mg respectively. Using mice as subject, the animal experiment confirmed that LABPG was non-toxic and safe. Effect of i.p. LABPG 0.5 mg/mouse on the phagocytic function were studied. It was suggested that the phagocytic activity of PM phi had markedly enhanced, the activity of serum lysozyme was increased significantly. YC-Rosette experiment suggested that the activity of C3b receptors of PM phi were increased and the YC-Rosette forming ratio were higher than controls. The difference was significant by statistical analysis. Therefore, it is considered that LABPG was able to activate M phi and improve immune function in mice. PMID- 12549403 TI - [Isolation and identification of Agrobacterium spp. from cherry crown galls and their sensitivities to agrocin 84]. AB - Crown galls were sampled from cherry yards of Shandong, Hebei, and Liaoning provinces. 46 pathogenic strains were isolated. Physiological and biochemical tests revealed that 4 strains were Agrobacterium tumefaciens(bio. 1) and that the other 42 strains were A. rhizogenes (bio. 2), according to the classifications and nomenclatures of the genus Agrobacterium and the species revised by Sawada et al and Bouzar. The Ti plasmids of all strains were nopaline type. All strains were sensitive to agrocin 84, which suggested that crown gall disease of cherry could be controlled by K84 strain. PMID- 12549404 TI - [Ultrastructures of a mycoplasma-like organism causing mulberry dwarf disease]. AB - The pathogen causing the dwarf disease of mulberry in Tongxiang, Zhejiang Province was investigated based on transmission electronic microscopy of the leaf vein of diseased plants collected in the field. A mycoplasma-like organism was identified in the sieve tube, companion, and parenchyma cells of the phloem tissues infected. The organism was spheric or oval in shape, ranging from 50 nm to 160 nm in diameter, and with bilamina unit membrane of 8-10 nm in thickness, and was filled with nuclear strands or fibrous material. Some nuclei of mesophyll cells in the diseased leaves were found disrupted as their nuclear membrane was broken, followed by of nucleo-cytoplasmic matrix and dispersive disappearance of nucleoli. Furthermore, other ultrastructural aberration including depletion of chloroplast stroma, disruption of chloroplast grana, increase of mitochondria and rough endoplasmic reticulum, and discomposition of some mitochondrial cristae were also observed associated with the pathological changes caused by the mycoplasma-like organism. In contrast, this type of organism was not found in healthy plants examined. Thus, the mycoplasma-like organism is likely to be a pathogen attributable to the mulberry dwarf disease. PMID- 12549405 TI - [Isolation and identification of Rhodopseudomonas acidophila from Fujian province, China]. AB - A strain P301 of Gram negative purple nonsulfur bacterium was isolated from the ditch's mud collected in Fuzhou, Fujian. The cells are rod-shaped. Multiplication occur by budding without stalk formation. Growth optimum pH is 5.0-5.7. No growth factors required. The cells contain bacteriochlorophyll a. The photosynthetic membrane system consists of parallel lamellae. According to Bergery's Manual of Determinative Bacteriology, 8th ed. (1974) and 9th ed. (1994), the strain was identified to be Rhodopseudomonas acidophila. But on the side of enzyme activity, utilization of organic substrates and others the strain some differ from the type strain described by N Pfennig (1969). Therefore the strain P301 was identified to be Rhodopseudomonas acidophila var. fujianensis n. var. PMID- 12549406 TI - [Establishment of hybridoma secreting anti-mycobacteria monoclonal antibody by using electrofusion technique]. AB - By using the electrofusion technique, the fusion of NS1 myeloma cells with spleen cells of BALB/c mouse immunized with five species of mycobacteria was carried out. With the optimal electrofusion conditions, we obtained a high fusion efficiency of 1.75 hybrid cell clones per 10(5) spleen cells. ELISA was used for screening secreting antimycobacteria McAb hybrid cells. After cloning, 22 stable hybrid cells lines were selected, and 8 of them had been studied. PMID- 12549408 TI - [Advance of research for microbial life in low temperature environments]. PMID- 12549407 TI - [Observations of characteristics of the mechanisms of biological oxidation of cell wall-deficient bacteria]. AB - The L-forms were induced from Staphylococcus aureus, Escherichia coli and Bacillus cereus by beta-lactam antibiotics and then observations on the properties of oxygen requirement, sugar fermentation and sensitive to cyanide of the L-forms were done. The results were shown that the L-forms derived from the obligate aerobe or the facultative anaerobe did not ferment sugars and were highly oxygen-dependent and more sensitive to cyanide than their parent bacteria. The metabolic activities which were same as the parent bacteria of the L-forms would return after the L-forms reverted to the typical bacteria forms. It was possible that the changes of biological oxidation mechanism were due to the deficiency of the cell wall which led to loss of the periplasmic space or the membrane-wall interlayer so that the enzymes for fermentation existed in the space could not be hold. PMID- 12549409 TI - [Introduction of the information on projects received and supported in division of microbiology NSFC in 1997]. PMID- 12549410 TI - [The composition and distribution of some kinds of anaerobic microorganisms in Yinqion basin]. AB - Under the strictly anaerobic conditions, the population of Sulfate-reducing bacteria, fermentative bacteria and methanogenic bacteria of serial samples got from erect sections of different Sedimentary of Yingqiong basin (a typical marine sedimentary environment) were measured by MPN method. The morphology of different kinds of bacteria and the metabolic types of methanogen and methanogenic activity were observed. The relation between population of bacteria and some indexes were compared. The results show that SRB present in all of the samples. The distribution of SRB and fermentative bacteria have no interrelation with the depth of samples but SRB has interrelation with the SO(4)2- concentration, and fermentative bacteria has negative interrelation with the contents of organic matter. Two kinds of methanogen present in all of the samples. They belong to Methanobacterium and Methanococcus, respectively. Their types of nutriment are H2/CO2. PMID- 12549411 TI - [Cloning of DNA sequences involved in exopolysaccharide synthesis of Xanthomonas campestris pv. campestris]. AB - By using a cloned DNA segment containing the mutated sequences from the exopolysaccharide-deficient mutant T117 of Xanthomonas campestris pv. campestris as a probe, a 9.4 kb HindIII fragment harbouring DNA sequences corresponding to the mutated site was identified and cloned from the wild-type strain. The fragment could complement the mutant strain T117 in trans and restore the synthesis of exopolysaccharide. This indicates that the fragment contains at least an intact gene which is involved in the exopolysaccharide synthesis of Xanthomonas campestris pv. campestris. PMID- 12549412 TI - [Characterization of the Streptomyces plasmid pSGL1]. AB - The plasmid pSGL1(7.4 kb) which was isolated from antitumor antibiotic C-1027 producing strain Streptomyces globisporus C-1027 was characterized. It could be introduced into Streptomyces lividans by transformation. Following a series of deletion experiments, the minimal replication region was located within a 2.0 kb Sau 3AI fragment. pSGL1 is compatible with pIJ101. pSGL1 and its derivatives have a copy number of 70-250. Some of the derived plasmids such as pSGLN and pSGLS3 have suitable selectable antibiotic resistance marker and unique restriction endonuclease sites and may be potentially useful as cloning vectors. These plasmids may be used to design and construct efficient secretory expression vectors. PMID- 12549413 TI - [Cloning and characterization of melanin biosynthesis genes from Streptomyces lincolnensis 78-11]. AB - Using mel C1-C2 gene from pIJ702 as probe to hybridize wity chromosome DNA of S. lincolnensis 78-11, a series of positive bands were found, including a 3.2 kb BanHI fragment and a 2.6 kb SphI fragment. So a gene library of S. lincolnensis, containg 3.0-3.5 kb BamHI fragment, was constructed. Melanin biosynthesis genes, mel C1 and mel C2, were then cloned and the fragment, inserted in recombinant plasmid pRSB336 were sequenced. Among five ORFs of the 3152 bp BamHI fragment, mel C1 and mel C2 were highly homologous to the respective genes of the other three strains of Streptomyces, morever, the product of mel C2 gene from S. lincolnensis 78-11 was slightly homologous to tyrosinase from human-being by 17.3% and from rat by 24.5%. All of the above results indicated that mel C1, mel C2 and orf 3 constitute the melanin biosynthesis operon of S. lincolnensis. In the further study of the cloned melanin biosynthesis genes of S. lincolnensis 78 11, recombinant plasmids, pPZ518 and pPZ519, which contain promoter of neo gene and mel gene in different orientation respectively, were constructed, and then were used to transform S. lividans TK23. 12 transformants of pPZ518, selected at random, secreted light brown pigment in R2YE culture medium, but all transformants of pPZ519 and pES1 were white. PMID- 12549415 TI - [Cloning and sequence analysis of rOmpA gene fragment of spotted fever group Rickettsiae isolated in China]. AB - Rr 190.70 p-602 n primers were used to amplify the rOmpA gene fragment of prototype strains (Rickettsia sibirica 246 strain and R. parkeri) and Chinese strains (BJ-90, Ha-91 and HLJ-054) of spotted fever group rickettsiae (SFGR). The PCR products were cloned into pGEM-T vector respectively and sequenced by Sanger's dideoxy method. The sequences of the tested strains were compared with the corresponding regions of prototype and reference strains of SFGR. The results showed that the homology of rOmpA gene fragments of prototype strains varied from 90.06%-96.62% in nucleotide and 83.05%-94.35% in putative amino acid respectively. Compared Chinese strains with prototype and reference strains of SFGR, it was found that the homology of BJ-90 strain and Ha-91 strain with R. sibirica 246 strain was the highest in nucleotide (99.06% and 98.31%, respectively) and in putative amino acid (98.87% and 96.61%, respectively), whereas HL-93 strain and HLJ-054 strain showed higher homology with R. japonica (96.62% and 95.68% in nucleotide, respectively) than others. Among all the Chinese strains, the homology between BJ-90 strain and Ha-91 strain was the highest, 99.25% in uncleotide and 97.74% in putative amino acid. The homology of nucleotide and putative amino acid between HL-93 strain and HLJ-054 strain was 98.12% and 94.92% respectively. The above results indicated that all SFGR so far isolated from different sources seem to be identical or closely related to R. sibirica, except for HL-93 and HLJ-054 strains, which could be considered as new members of SFGR. PMID- 12549414 TI - [Subcloning and expression of coding region for cellulase binding domain of CBH I from P. janthinellum in E. coli]. AB - The in vitro DNA manipulations, included the nested deletions, of cbh1 from P. janthinellum inserted into pUC18-181 were carried out. The two ends of fragments were modified into blunt ends and the fragments were self-ligated. Then, the encircled plasmids were transformed to E. coli JM109. Utilizing the characterization of CBD binding to crystalline cellulose, one catalytic domain deletion transformant producing active LacZ-CBD fusion protein was isolated from 24 transformants randomly picked from 400 transformants. The molecular weight of the LacZ-CBD fusion protein is 21 kD. The plasmid was designated pUC 18C. The LacZ-CBD fusion protein produced by JM109(pUC18C) was able to be purified by procedure of adsorption-desorption to cellulose. The pNPC activity of crude enzyme solution of JM109(pUC18C) induced by IPTG were zero, identified the JM109(pUC18C) has no CBHI activity. PMID- 12549417 TI - [Isolation, purification and identification of metallthionein from strain BD 101 of Saccharomyces cerevisiae]. AB - Selection of Saccharomyces cerevisiae BD101 having resistance to metal ions and producing metallothionein by inducer from Saccharomyces. These proteins were isolated and purified by Sephadex G-50 and subsequent DEAE Sepharose CL-4B, then Sephadex G-25 for desalination. There were two iso-MTs for Cu-induced. The, molecular weights were 7 kD and had 60 amino acids. Both Cu-MTs were rich in Cysteine (10%). 4 atom Cu/6 mole Cys/mole Cu-MTs. PMID- 12549416 TI - [Analysis of higher basidiomycetous mitochondrial DNA with four-GC-cutter restriction enzymes]. AB - The mitochondrial DNA band patterns emerged from basidiomycetous genomic DNA digested with four-GC-cutter restriction enzymes, HaeIII, CfoI and MspI, were reported in this paper. It was demonstrated that mtDNAs of higher basidiomycetes had abundant diversity after comparison of their band patterns, summated sizes, the number of fragments and homology among 12 strains of Agaricales, Aphyllophorales. The results also showed that the technique, displaying fungal mtDNA bands directly from their total DNA, would be more efficient and sensitive for analyses of fungal mtDNA RFLP if several four-GC-cutter restriction enzymes, instead of one, were used. PMID- 12549418 TI - [Studies on the yeasts and yeast-like fungi degrading trinitrotoluene]. AB - Seventeen strains of yeasts and yeast-like fungi were isolated from soil and waste water samples polluted by trinitrotoluene (TNT). These strains can degrade 71%-93% TNT when growing in the medium originally containing 70-80 mg/L TNT within 40 h. They were identified. Among them, six strains are Candida krusei, four are C. quercitrusa, one is C. famata, one is Hansenula beijerinckii, one is H. subpelliculosa, and four are Geotrichum candidum. Six strains were selected for further studies on the conditions effecting TNT degradation by them. The optimum pH and temperature are pH 7 and 37-40 degrees C, respectively. TNT Degradation ability of the strains can be promoted by adding 0.01%-0.05% glucose or 0.01%-0.1% yeast extract into the medium. Addition of 0.05% (NH4)2SO4 or NH4Cl to the medium can evidently inhibit the degradation of TNT by the strains. PMID- 12549419 TI - [Enzymatic synthesis of cephradine]. AB - Cephradine was synthesized by gamma-alumina-immobilized form of the penicillin G acylase of Bacillus megaterium with D-phenyglycine methylester hydrochloride (CH DGME.HCl) as acyl donor and 7-aminodeacetoxycephalosporanic acid (7-ADCA) as acyl acceptor. 0.1 g of 7-ADCA was dissolved by adding 2.5 ml of distilled water and about 0.25 ml of 2 mol/L NaOH in a 25 ml flask. To the solution, after 0.25 g of CHDGME.HCl was added, 0.1 mol/L phosphate-0.05 mol/L citric acid buffer, pH 7.5 was added to result in a volume of 5 ml with pH 7.5 Then 1 g(220 IU) of immobilized enzyme was added. The flask was shaken on a rotary shaker at 110 r/min and 25 degrees C for 5 h. The conversion rate of 7-ADCA was 81%. In an expanded experiment in 500 ml of reactive volume, 11.8 g of cephradine was obtained from 10 g of 7-ADCA. The conversion rate of 7-ADCA was 80% with about 87% yield of cephradine. Enzymatic synthesis was inhibited in varying degrees by phenylacetic acid, phenoxyacetic acid and cephalosporin G. PMID- 12549420 TI - [Appllcation of PYGC and the hierarchical clustering analysis to recognization of viruses]. AB - The pyrolysis gas chromatography of 29 strains nuclear polyhedrosis viruses, 11 strains cytoplasmic polyhedrosis viruses and 16 strains granulosis viruses were performed on a shimadzu GV-9A capillary gas chromatography equipped with a pyrolyser model PYR-2A. The differences among the inclusion bodies of NPV, CPV and GV could clearly be distinguished and also beidentified through analysis of fingerprinting the hierarchical clustering analysis of GC, This procedure has provided a usefel method for the classification and identification of viruses. PMID- 12549421 TI - [Purification and properties of alkaline cellulase from alkalophilic Bacillus N6 27]. AB - The alkaline cellulase produced by alkalophilic Bacillus sp. N6-27 was purified to electrophoresis homogeneity by (NH4)2SO4 fractionation, Sepharose CL-4B hydrophobic interaction chromatography, Bio-gel P-150 chromatography. The molecular weight and pI determined by SDS-PAGE and by PAGE-IEF were 94,000 and 4.2, respectively. The optimum temperature and pH for the enzymatic catalysis were 55 degrees C and 8.5, respectively. The enzyme activity was stable under 50 degrees C and in the pH range of 6-11. The substrate was carboxymethylcellulose (CMC). The enzyme activity was strongly inhibited by Fe2+, Cu2+ and Hg2+. PMID- 12549422 TI - [Isolation of a lipase-producing Pseudomonas strain and optimization of its fermentation conditions]. AB - A lipase-producing bacterium strain was isolated from soil and was identified as Pseudomonas sp.. Its lipase yield was improved 2.25-fold by combined treatment of UV irradiation and NTG. The lipase fermentation condition for the mutant strain was optimized with Plackett-Burman design and Response Surface Analysis (RSA), and the formula of the optimum medium suitable for industrial scale fermentation was thereby established. A maximum yield of 87.5 U/ml was obtained. PMID- 12549423 TI - [Screening and breeding of highly-effected degrading cotton-phenol strains and study on defoxication technology and conditions]. AB - From mildewed cottonseed cake and stock cultures of mold and yeast. We select more than ten strains of yeasts and molds which can degrade cotton phenol. At last we got four strains which can degrade cotton phenol highly effected after mutagenized by physical and chemical factors and induced by cotton phenol. They belong to Candida tropicalis, Torulopsis candida, Aspergillus flavus and Aspergillus niger. By small and medium size fermenfations, the content of dissociated cotton phenol all reach safe criterion (the lowest dissociated cotton phenol content is 220 mg/L in the defoxicated cottonseed cake), compared with FeSO4 method, defoxicated cottonseed cake looks and tastes well and the content of protein and amino acid are highly enhanced as will. PMID- 12549424 TI - [The comparison on growth rate and photopigments of filaments of Spirulina platensis strain Z with different morphology]. AB - The line-shaped filaments Sp-Z(L) were isolated and cultured from Spirulina platensis Sp-Z. The growth rate of Sp-Z(L) was only 64% as much as that of Sp-Z when the light intensity was 4000 Lux. The contents (x 10(-3) g/g dry weight) of chlorophylls, carotenoids and phycobilins of Sp-Z(L) and Sp-Z were 20.6, 0.343, 5.00 and 24.1 0.297, 4.46, respectively. Moreover, as to the absorption spectra of the three photopigments of Sp-Z(L), red shifts were observed. Therefore, after the spiral Sp-Z breeded or changed into Sp-Z(L), The contents of carotenoids and phycobilins raised 23% and 12%, respectively, but the contents of chlorophylls dropped 14.5%, and the growth rate dropped 36% too. The results probably showed that the decrease of chlorophylls was one of the main reasons that caused the remarkable decrease of the growth rate of Sp-Z(L). PMID- 12549425 TI - [Advance of molecular biological research of filamentous pathogenic fungi]. PMID- 12549426 TI - [Proceedings of the 11th Session of the General Meeting of the Russian Academy of Medical Sciences on Medical Problems of Biosafety. April 3-6, 2002, Moscow]. PMID- 12549427 TI - [Structure determination of alkyd resins by simultaneous pyrolysis methylation gas chromatography-mass spectometry]. AB - Several types of alkyd resins have been analyzed by simultaneous pyrolysis methylation gas chromatography mass spectrometry (SPM-GC-MS). The samples were mixed with tetramethylammonium hydroxide, and the process included simultaneous pyrolysis derivatization and mass spectrometry determination. SPM-GC-MS conditions: a vertical microfurnace-type pyrolyzer was directly attached to a gas chromatograph-mass spectrometer. About 20 ug of the mixed alkyd resin sample was introduced into the center of the pyrolyzer at 450 degrees C under the flow of helium carrier gas. The inlet mode was split with an injection interval of 10 s and the inlet pressure was 82.7 kPa. The gas chromatograph was fitted with a fused-silica capillary column ( SE-54 30 m x0.25 mm i.d. x 0.25 um). Temperature program settings were: initial temperature, 40 degrees C, hold 2 min; increase at 6 degrees C min 1 to 220 degrees C, hold 14 min; increase at 10 degrees C min 1 to 280 degrees C, hold 8 min. All pyrolysis products referred to were identified by mass spectrometry. The temperature of the ion source was 210 degrees C and that of the transfer line was 250 degrees C. The technique could give additional information about the composition of the dibasic acids, polyols, six types of drying oils, and modified alkyd resins. The method is sensitive, accurate, convenient, and involves minimal sample manipulation. PMID- 12549428 TI - [Scientific-practical conference with the international participation "Tuberculosis is an old problem in the new millennium". Novosibirsk, July 1-5, 2002]. PMID- 12549429 TI - [Old and new prescriptions for infectious diseases and the newest recipes for biomedical products in plants]. AB - The three antiviral vaccines discovered in the 18th century (smallpox), 19th century (rabies), and 20th century (polio) share a common feature: none would ever be licensed today for human vaccination. Yet Jenner's smallpox vaccine led to the eradication of smallpox, Pasteur's rabies vaccine represented the first successful post-exposure treatment of people bitten by rabid animals, and polio vaccine administered since its discovery in 1950 is leading to the eradication of polio (in the years 2004-2005) from the earth. However, in the case of rabies, efforts at complete eradication are unrealistic, despite the availability of a very effective vaccine, since rabies, unlike smallpox and polio, is not limited to humans and can infect all domestic and wild mammalian species. Rabies is probably the oldest known infectious disease, yet knowledge of the virus and the disease is far from complete. For instance, the appearance of 24 cases of "cryptic" rabies in the USA, i.e. cases not associated with any bite or scratch, with an incubation period in humans extending 6-8 years, is a puzzling phenomenon that cannot be readily explained. On the other hand, rabies is one of the few strictly neuronal infections and, as such, is an excellent model for the study of neurotropic virus distribution in the brain. Apoptosis induced by a rabies strain expressing high levels of glycoprotein spreads much more slowly through brain tissue than that induced by strains producing lower glycoprotein levels. Attenuated rabies virus constructed to express twice the normal glycoprotein levels is also an excellent antigen for induction of immune responses in the host. Foreign antigens using this vector may also produce highly immunogenic vaccines. Global approach to immunization. Those monitoring the spread of AIDS in many parts of the world know that cost of treatment is one of the major problems in combating the disease. Vaccines against HIV face the same problem. In general, the price of vaccines and sera is exorbitant for the afflicted population in developing countries. In addition, the dearth of syringes, the unavailability of nurses and doctors to administer multiple vaccine injections, and other factors in these countries require a drastic change in current vaccine production approaches. About 12 years ago, plants became vehicles to produce biomedical reagents. Plants can be exposed directly to a construct containing a foreign gene and Agrobacterium to create a transgenic plant that, over several generations, produces the desired product. Alternatively, plants infected with a plant virus (e.g. alfalfa mosaic virus) fused with a foreign gene can propagate the foreign antigen as the virus multiplies. Extraction of the plant virus followed by purification provides the desired biomedical product. Our use of either of these systems has led to the creation of plants producing vaccines, sera, hormones, and other biological reagents. In two clinical trials at the Institute of Bioorganic Chemistry of the Polish Academy of Sciences in Poznan (Poland), volunteers who ingested lettuce expressing hepatitis B vaccine showed hepatitis B antibodies in their sera. In another trial carried out at the Biotechnology Foundation Laboratories in Philadelphia (USA), volunteers ingesting a spinach-rabies vaccine showed an immunological priming effect, since only one injection of commercially available rabies vaccine significantly raised the level of rabies-specific antibodies. Vaccines against HIV gp120 and Tat have been produced in spinach, and a construct of gp120 with the CD4 receptor is now being adapted to this plant. Two types of antibodies against rabies and against colorectal cancer are being produced in tobacco and in lettuce. The suboptimal quality of the currently available anthrax vaccine prompted our efforts to produce the anthrax protective antigen (PA) in tobacco and lettuce. Quite clearly, plants will play a prominent role in producing a variety of biomedical reagents in the future. PMID- 12549430 TI - The Australasian Triage Scale. PMID- 12549431 TI - Standard terminology. PMID- 12549432 TI - Changing prescription software to favour generics could save Australia 40m pounds sterling a year. PMID- 12549433 TI - States organise joint non-profit prescription plan to cut drug costs. PMID- 12549434 TI - Study reports on use of "touch screen" health kiosks. PMID- 12549435 TI - Learning disability task force criticises draft mental health bill. PMID- 12549436 TI - NHS and local government try to stop introduction of "bed blocking" fines. PMID- 12549437 TI - Commentary: Improve the quality of the consultation. PMID- 12549438 TI - Commentary: Clinical focus might make it work. PMID- 12549439 TI - Doctors and computers. . . . While limited and uncoordinated development of information technology curtails general practitioners. PMID- 12549441 TI - Rethinking management. Consistent criteria need to be applied to rationing decisions. PMID- 12549443 TI - Long term effects of advice to reduce dietary salt. Critical faculties should always be exercised. PMID- 12549444 TI - Long term effects of advice to reduce dietary salt. Salt needs to be reduced in manufacturing and processing food. PMID- 12549445 TI - USDOT releases 2001 highway fatality statistics. PMID- 12549446 TI - Indian objection to export of human tissue for research. Problem is not that simple. PMID- 12549447 TI - NHTSA reports increased seat belt use by drivers, passengers. PMID- 12549448 TI - EPA raises bar for listing hazardous chemicals as bioaccumulative. PMID- 12549449 TI - Necessary evil? PMID- 12549450 TI - Your country needs you. PMID- 12549451 TI - When two tribes go to war. PMID- 12549452 TI - Patient information. What is carpal tunnel syndrome? PMID- 12549453 TI - Generation game. PMID- 12549454 TI - Total control: Roger Gosden interviewed by Alison Motluck. PMID- 12549455 TI - The accused. PMID- 12549456 TI - Taking the plunge. PMID- 12549457 TI - Free for all. PMID- 12549458 TI - The revolution has begun. PMID- 12549459 TI - Would you trust these people with your genes? PMID- 12549460 TI - One small step for lab rats. PMID- 12549461 TI - JAMA patient page. Men and women are different. PMID- 12549462 TI - Lab rats, rejoice! PMID- 12549463 TI - A wild gene chase. PMID- 12549465 TI - The genie is out. PMID- 12549466 TI - The right to choose. PMID- 12549468 TI - Disaster in the making. PMID- 12549471 TI - We're all suspects now. PMID- 12549472 TI - The DNA police. PMID- 12549473 TI - Analgesic prescribing. PMID- 12549474 TI - Meddling with monkeys. PMID- 12549475 TI - Too close for comfort. PMID- 12549476 TI - Animal madness. PMID- 12549477 TI - Trial and error. PMID- 12549478 TI - Fetal cell transplants: special report. PMID- 12549479 TI - Endocarditis associated with body piercing. PMID- 12549480 TI - Early onset brain tumor and lymphoma in MSH2-deficient children. PMID- 12549483 TI - Hereditary p16-Leiden mutation in a patient with multiple head and neck tumors. PMID- 12549484 TI - Another case of imprinting defect in a girl with Angelman syndrome who was conceived by intracytoplasmic semen injection. PMID- 12549485 TI - From the Centers for Disease Control and Prevention. Laboratory-acquired West Nile virus infections--United States, 2002. PMID- 12549486 TI - From the Centers for Disease Control and Prevention. Global progress toward laboratory containment of wild polioviruses--July 2001-August 2002. PMID- 12549488 TI - Human T-cell lymphotropic virus type 1: is it time to screen organ donors and recipients? PMID- 12549489 TI - Innominate artery transection secondary to blunt trauma. PMID- 12549487 TI - From the Centers for Disease Control and Prevention. Measles outbreak among internationally adopted children arriving in the United States, February-March 2001. PMID- 12549490 TI - Transected axillary artery: unusual blunt trauma. PMID- 12549491 TI - Successful re-implantation of retrieved large segment of open femoral fracture. PMID- 12549499 TI - Recipient of the 2003 Pioneer Award. Keith J Betteridge, B.V.Sc, M.V.Sc., Ph.D. PMID- 12549501 TI - Health insurance premiums rose more than 30% from 1996 to 2000. PMID- 12549502 TI - Initiative to increase physician awareness of racial/ethnic disparities in cardiac care. PMID- 12549500 TI - How theories became knowledge: Morgan's chromosome theory of heredity in America and Britain. AB - T. H. Morgan, A. H. Sturtevant, H. J. Muller and C. B. Bridges published their comprehensive treatise The Mechanism of Mendelian Heredity in 1915. By 1920 Morgan's "Chromosome Theory of Heredity" was generally accepted by geneticists in the United States, and by British geneticists by 1925. By 1930 it had been incorporated into most general biology, botany, and zoology textbooks as established knowledge. In this paper, I examine the reasons why it was accepted as part of a series of comparative studies of theory-acceptance in the sciences. In this context it is of interest to look at the persuasiveness of confirmed novel predictions, a factor often regarded by philosophers of science as the most important way to justify a theory. Here it turns out to play a role in the decision of some geneticists to accept the theory, but is generally less important than the CTH's ability to explain Mendelian inheritance, sex-linked inheritance, non-disjunction, and the connection between linkage groups and the number of chromosome pairs; in other words, to establish a firm connection between genetics and cytology. It is remarkable that geneticists were willing to accept the CTH as applicable to all organisms at a time when it had been confirmed only for Drosophila. The construction of maps showing the location on the chromosomes of genes for specific characters was especially convincing for non-geneticists. PMID- 12549503 TI - Study finds that TV medical dramas address health policy issues. PMID- 12549504 TI - Toxic chemicals website. PMID- 12549505 TI - A matter of standards: the quest for authentic, reproducible and reliable plant drugs. PMID- 12549506 TI - The town of St. Gall and its cathedral. PMID- 12549507 TI - A century and a half of pharmacy in Trumpington Street, Cambridge. PMID- 12549509 TI - Fatherless children and Accouchement sous X, from marriage to demarriage: a paradigmatic approach. AB - The author proposes a paradigmatic approach to the construction of fatherless, abandoned, and/or motherless children as social issues needing some policy responses: within the different historical Western configurations of the repertoire of potential responses, major organizing paradigms draw the line between those responses that are rejected as belonging to the supreme evil sphere and others that can be considered as lesser evils. Within the framework of a paradigm, dilemmas exist and debates go on as to which lesser evils should be preferred and how the policies should be implemented. Policies deriving from different paradigms give rise to intractable controversies since the disagreements concern not only the means but also the ends. Four paradigms can be extracted from the historical data with regard to illegitimacy and single motherhood: Christian angelism, Malthusian angelism, healthy citizenship, and proper citizenship. PMID- 12549510 TI - Plants against malaria, part 2: Artemisia annua (Qinghaosu or the sweet wormwood). PMID- 12549511 TI - [Genetic mutation in head and neck squamous cell carcinoma]. PMID- 12549512 TI - [Foreign bodies of trachea and esophagus in children]. PMID- 12549513 TI - Proceedings of the German Conference on Bioinformatics. 7-10 October 2001, Braunschweig, Germany. PMID- 12549514 TI - Bioinformatics research and education in Germany. PMID- 12549515 TI - Bioinformatics service, education and research: the EMBnet and CBI. European Molecular Biology Network Centre of Bioinformatics. PMID- 12549516 TI - Invasive pneumococcal disease in Australia, 2001. AB - There were 1,681 cases of invasive pneumococcal disease (IPD) notified to the National Notifiable Diseases Surveillance System in Australia in 2001; a rate of 8.6 cases per 100,000 population. The notification rate varied between states and territories and by geographical region with the highest rates in the north of the country. Pneumococcal disease was reported most frequently in children aged less than 5 years (47.3 cases per 100,000 population). Enhanced surveillance for IPD was carried out in the Northern Territory, Western Australia, South Australia, Victoria, Tasmania and metropolitan areas of New South Wales, encompassing 72 per cent of the population and providing additional data on 86 per cent of all notified cases. Enhanced surveillance data revealed high rates of pneumococcal disease in Indigenous Australians. Rates of IPD in Indigenous children aged less than 5 years were as high as 483 cases per 100,000 population in the Northern Territory. The clinical presentation of IPD was most commonly pneumonia (56%) and bacteraemia (36%). There were 125 deaths attributed to IPD resulting in an overall case fatality rate of 8.6 per cent. More than half (54%) of all cases had a recognised risk factor for IPD. Eighty-six per cent of serotypes identified in non-indigenous children compared with only 55% of serotypes in Indigenous children were in the 7-valent vaccine. Antibiotic susceptibility testing showed reduced susceptibility to penicillin in 12 per cent, and to third generation cephalosporins in 5 per cent of isolates. These are the first national data available on IPD in Australia and will assist in evaluating the impact of the newly introduced conjugate vaccine and guide overall pneumococcal vaccine strategies. PMID- 12549517 TI - Invasive pneumococcal disease in north Queensland, 2001. AB - This report provides information on the 93 locally-acquired cases of invasive pneumococcal disease (IPD) notified in children and adults in north Queensland in 2001. Indigenous people represented 38 (41%) cases. Almost half (45) of all cases were in children under 15 years of age, 20 (44%) of these were in children less than 2 years of age and 20 (44%) in Indigenous children. Five severe cases of IPD occurred, all in non-indigenous children under 2 years of age. Nine (10%) of the isolates from cases, mainly in young children, had some level of resistance to penicillin. Pneumococcal vaccination programs (including the Indigenous 'elderly and at-risk' adult program and the paediatric 'Indigenous and medically at-risk' conjugate vaccine program) are in place in Queensland although the vaccine is not currently funded for other at-risk groups. If vaccine recommendations had been adhered to in a timely fashion, two of the cases in children and one third (16) of the cases in adults that occurred in 2001 could potentially have been prevented. PMID- 12549518 TI - Tuberculosis notifications in Australia, 2001. AB - In 2001, there were 997 cases of tuberculosis (TB) reported to the National Notifiable Diseases Surveillance System, of which, 967 were new cases of TB and 30 cases were relapses. The incidence rate of TB in Australia in 2001 was 5.1 cases per 100,000 population. The highest incidence of TB was reported in people born overseas (19.3 cases per 100,000 population), followed by Indigenous Australians (9.8 cases per 100,000 population). In contrast, the incidence rate of TB in the non-Indigenous Australian-born population was 1.0 cases per 100,000 population. This pattern of TB incidence rates amongst the sub-populations of Australia has been observed for over 10 years. Eighty-six per cent of TB cases completed treatment in 2001. Treatment was unsuccessful in 7 cases and only 22 cases defaulted. The National Tuberculosis Advisory Committee has published a National Strategic Plan with performance indicators to ensure that this enviable record of TB control is maintained and improved. PMID- 12549519 TI - Report of the Australian Rotavirus Surveillance Program, 2001/2002. AB - The National Rotavirus Reference Centre together with collaborating laboratories Australia-wide has conducted rotavirus surveillance since June 1999. The serotypes of rotavirus strains that are responsible for the hospitalisation of children with acute gastroenteritis were determined for the period 1 June 2001 to 31 June 2002. We examined 754 rotavirus samples using a combination of monoclonal antibody immunoassay, reverse transcription-polymerase chain reaction, and Northern hybridisation. For the first time, serotype G9 strains were the most prevalent type nationally (40.4%) and found in 8 of the 9 centres. Serotype G1 strains were the second most prevalent type (38.9%), identified in 5 of the centres. These findings have important implications for vaccine development strategies which target serotypes G1-G4. PMID- 12549520 TI - Surveillance of antibiotic resistance in Neisseria gonorrhoeae in the WHO Western Pacific Region, 2001. World Health Organization. AB - A long-term program of surveillance of antimicrobial resistance in Neisseria gonorrhoeae isolated in the World Health Organization's Western Pacific Region (WHO WPR GASP) continued in 2001. Seventeen focal points contributed data on about 10,000 gonococci. Resistance to quinolone and penicillin antibiotics remained widely dispersed and at high levels. Gonococci with decreased susceptibility to third generation cephalosporins were again observed in four centres. Spectinomycin resistance was infrequently encountered. Control of gonorrhoea in the WHO WPR is compromised by the further reduction in options for cheap and effective treatment of gonorrhoea. PMID- 12549521 TI - OzFoodNet: enhancing foodborne disease surveillance across Australia: quarterly report, April to June 2002. PMID- 12549522 TI - Norwalk-like virus--issues for surveillance. PMID- 12549523 TI - Norwalk-like virus outbreak in Canberra: implications for infection control in aged care facilities. AB - This paper reports on an outbreak of viral gastroenteritis in three institutions (two aged care facilities and one hospital) in Canberra during the winter of 2002. Norwalk-like virus genotype II was detected in samples from staff and/or residents in all three institutions. A case series investigation was conducted amongst both staff and residents. It is likely that the outbreaks in the three institutions were linked due to transfers of infected residents from one institution to another, early in the outbreak. A total of 281 cases were identified during the outbreak, which lasted 32 days. Attack rates in the three institutions were 46.3 per cent, 52.7 per cent and 55.2 per cent respectively. Person-to-person spread and/or airborne transmission were postulated as modes of transmission in all three institutions. Infection control practices in each of the aged care institutions were of an acceptable standard for accreditation, but were inadequate to control further spread of the outbreak within and between institutions. Outbreak management plans should be a part of the infection control standards for accreditation of aged care facilities. PMID- 12549524 TI - An outbreak of infections with a new Salmonella phage type linked to a symptomatic food handler. AB - In December 2001, the South Australian Communicable Disease Control Branch investigated an outbreak of gastrointestinal illness linked to a Korean style restaurant in metropolitan Adelaide. Twenty-eight people were identified as having experienced gastrointestinal symptoms subsequent to dining at the restaurant between 9 and 12 December 2001. A case-control study implicated mango pudding dessert (OR 16.67 95% CI 2.03-177.04) and plain chicken (OR 10.67 95% CI 1.04-264.32). Nineteen cases and one food handler submitted faecal specimens that grew Salmonella Typhimurium 64var. Two samples of mango pudding and one sample of pickled Chinese cabbage also grew Salmonella Typhimurium 64var. The infected food handler reported an onset of illness 2 days before cases first reported eating at the restaurant. The food handler's only role was to prepare the mango pudding dessert in an area external to the restaurant's kitchen. Illness was strongly associated with consumption of a contaminated mango pudding dessert, with contamination most likely resulting from the symptomatic and culture positive food handler who prepared the dish. This outbreak demonstrates the importance of excluding symptomatic food handlers, and the need for appropriately informing and educating food handlers regarding safe food handling procedures. Restaurants with staff and management from non-English speaking backgrounds should be specifically targeted for education that is both culturally sensitive and language specific. PMID- 12549525 TI - A statewide outbreak of Salmonella bovismorbificans phage type 32 infection in Queensland. AB - Between 30 May and 1 June 2001, 10 cases of Salmonella Bovismorbificans infection were reported to Public Health Services, Queensland Health. Investigations included enhanced surveillance, case interviews, a matched case control study, environmental audit and microbiological testing of faecal isolates (phage typing) and implicated food products. Forty-one cases of S. Bovismorbificans infection were detected, 36 cases were phage type 32. A matched case control study identified that illness was associated with consumption of food from 15 outlets of a fast food chain, Company A (matched odds ratio [MOR] 17.5, 95% CI 2.0-657.3, p = 0.004) and consumption of a particular product, Product X (MOR undefined, p < 0.001) in the week before onset of illness. Manufacturers of Product X ingredients were audited. Deficiencies were identified in equipment cleansing at the salad mixture processing plant (Manufacturer M). A swab of food residue behind the cutting wheel rim of the lettuce shredder was positive for S. Bovismorbificans phage type 32. This appears to be the first reported Australian outbreak of salmonellosis associated with a lettuce product. The investigations suggest that inadequate maintenance of cutting equipment to prepare lettuce ingredients for Product X by Manufacturer M was a key factor in this statewide outbreak. The statewide nature of this outbreak demonstrates the role of timely serovar identification of Salmonella isolates by a reference laboratory as an aid to outbreak identification, and the importance of adherence to appropriate food safety procedures in the manufacture and preparation of mass produced food items for the public. PMID- 12549526 TI - Outbreak of staphylococcal enterotoxin food poisoning. PMID- 12549527 TI - Surveillance of viral pathogens in Australia--varicella-zoster virus. PMID- 12549528 TI - Using the Australian Childhood Immunisation Register to track the transition from whole-cell to acellular pertussis vaccines. AB - From 1997 to 1999, Australia changed from a whole-cell based pertussis vaccination program to an acellular one. This paper tracks the transition from whole-cell to acellular pertussis vaccines by calculating the number of whole cell (DTPw) and acellular (DTPa) pertussis vaccines recorded on the Australian Childhood Immunisation Register (ACIR) each month from January 1996 to August 2000. The number of combined diphtheria-tetanus (CDT) vaccines, recommended where DTP is contraindicated and for the fifth dose prior to 1994, was also calculated. The use of DTPa increased following its licensing in 1997, with a corresponding decrease in the use of DTPw. The increase was initially greatest in its use as a fourth and fifth dose, for which it was funded at a national level in 1997. Subsequently, a steep increase in its use for the first three doses followed in 1999, coinciding with it becoming free of charge for infants nationally. The use of CDT has decreased markedly since January 1996 and, since March 2000, fewer than 100 CDT vaccines per month were recorded on the ACIR, suggesting that this vaccine is not being inappropriately used. PMID- 12549529 TI - Infection control and public health aspects of a case of pertussis infection in a maternity health care worker. PMID- 12549530 TI - An unusual reaction to rabies vaccine. PMID- 12549531 TI - A measles outbreak in the Whitsundays, Queensland: the shape of things to come? AB - This report describes a small outbreak of measles that occurred in the Whitsunday region, north Queensland, in July to August 2002. With one exception, all the cases were deliberately unvaccinated because their parents were conscientious objectors to vaccination. It is suggested that this pattern of measles outbreaks, with most cases being not preventable because of conscientious objection, will become increasingly recognised in the future. PMID- 12549532 TI - Refining the public health response to primary meningococcal conjunctivitis. AB - Primary meningococcal conjunctivitis (PMC) is accepted as an uncommon condition. This report describes two recent cases of PMC in newborn infants in a hospital nursery. In both cases the organisms identified were non-groupable strains of N. meningiditis, considered to be of low pathogenic potential. Both infants received systemic therapy and recovered without sequelae. The Guidelines for the early clinical and public health management of meningococcal disease in Australia recommend the notification of PMC to public health authorities and chemoprophylaxis of contacts. However, our 2 cases suggest that the guidelines should allow for an assessment of risk in determining the public health response. This assessment should include the severity of the conjunctivitis and the serogroup of the N. meningitidis isolate. PMID- 12549533 TI - A focal, rapidly-controlled outbreak of dengue fever in two suburbs in Townsville, north Queensland, 2001. AB - In April-May 2001 an outbreak of dengue fever occurred in two suburbs in Townsville, north Queensland. This was the first outbreak in the Townsville region since a very large outbreak in 1992-1993. Notification delays resulted in late detection of the outbreak. Once recognised, control measures were implemented and rapid control was achieved. Dengue serotype 2 was the causative virus and 9 cases of dengue fever were documented. The approach to management of dengue fever outbreaks and vector control strategies have been improved and refined in the years since the 1992-1993 outbreak. These measures, in addition to favourable weather conditions, were likely to have contributed to the successful containment of this outbreak. PMID- 12549534 TI - An outbreak of Barmah Forest virus disease in Victoria. AB - This report describes the epidemiological and clinical features of an outbreak of 47 cases of laboratory-confirmed Barmah Forest virus disease (BF disease) that occurred in Victoria between January and May 2002. Laboratory-confirmed cases were investigated, and information on travel history and clinical details was collected. Surveillance data from adult mosquito trappings and climatic conditions in the Wellington Shire were also reviewed. The response rate for interviews was 85 per cent (40/47). The most common symptoms reported by cases included arthralgia (95%), lethargy (90%) and maculopapular rash (72.5%). Transmission of BF disease in the Gippsland region was associated with unusually high numbers of Ochlerotatus camptorhynchus mosquitoes. This outbreak was of interest due to the fact that cases of BF disease outnumbered cases of Ross River virus disease (RR disease) in Victoria for the first time since data were available. Similar outbreaks of BF disease, in the absence of RR disease, occurred in Western Australia in 1993 and New South Wales in 1994/1995. Although the majority of BF disease cases reported regular outdoor activity during which they could be exposed to mosquito populations, they infrequently take precautions to limit exposure. Further efforts need to be made to educate people of the importance of using repellents and other personal preventative measures. PMID- 12549535 TI - The Australian Institute of Health and Welfare. AB - This overview has described the AIHW role in Australia's health information which is highly relevant to communicable disease surveillance. The AIHW provides statistics and information on the nation's health and welfare within local, state, national and international settings. It has established an expertise in the collection, standards and dissemination of information. Given these qualities, the AIHW has a wealth of expertise and welcomes the use of its publications and data sources. Details of the Institute's work can be found on its website (www.aihw.gov.au). PMID- 12549536 TI - Communicable diseases surveillance. PMID- 12549537 TI - Laboratory-scale measurement of trace gas fluxes from landfarm soils. AB - Trace gas emissions from refinery and bioremediation landfarms were investigated in a mesocosm-scale simulator facility. Five simulators were constructed and integrated with a data acquisition system and trace gas analyzers, allowing automated real-time sampling and calculation of total hydrocarbon (THC), CO2, and water vapor fluxes. Experiments evaluating the influence of simulated cultivation and rainfall on trace gas fluxes from the soil surfaces were conducted. Results were compared with published field results. Results showed that cultivating dry or moderately wet soil resulted in brief enhancements of THC fluxes, up to a factor of 10, followed by a sharp decline. Cultivating dry soil did not enhance respiration. Cultivating wet soil did result in sustained elevated levels of respiration. Total hydrocarbon emissions were also briefly enhanced in wet soils, but to a lesser magnitude than in dry soil. Hydrocarbon fluxes from refinery landfarm soil were very low for the duration of the experiments. This lead to the conclusion that elevated THC fluxes would only be expected during waste application. An evaluation of the influence of simultaneous water vapor fluxes on other trace gas fluxes highlighted the importance in lab-scale experiments of correcting trace gas fluxes from soils. The results from this research can be used to guide management practices at landfarms and to provide data to aid in assessing the effect of landfarms. PMID- 12549538 TI - Greenhouse gas emissions and soil indicators four years after manure and compost applications. AB - Understanding how carbon, nitrogen, and key soil attributes affect gas emissions from soil is crucial for alleviating their undesirable residual effects that can linger for years after termination of manure and compost applications. This study was conducted to evaluate the emission of soil CO2, N2O, and CH4 and soil C and N indicators four years after manure and compost application had stopped. Experimental plots were treated with annual synthetic N fertilizer (FRT), annual and biennial manure (MN1 and MN2, respectively), and compost (CP1 and CP2, respectively) from 1992 to 1995 based on removal of 151 kg N ha(-1) yr(-1) by continuous corn (Zea mays L.). The control (CTL) plots received no input. After 1995, only the FRT plots received N fertilizer in the spring of 1999. In 1999, the emissions of CO2 were similar between control and other treatments. The average annual carbon input in the CTL and FRT plots were similar to soil CO2-C emission (4.4 and 5.1 Mg C ha(-1) yr(-1), respectively). Manure and compost resulted in positive C and N balances in the soil four years after application. Fluxes of CH4-C and N2O-N were nearly zero, which indicated that the residual effects of manure and compost four years after application had no negative influence on soil C and N storage and global warming. Residual effects of compost and manure resulted in 20 to 40% higher soil microbial biomass C, 42 to 74% higher potentially mineralizable N, and 0.5 unit higher pH compared with the FRT treatment. Residual effects of manure and compost on CO2, N20, and CH4 emissions were minimal and their benefits on soil C and N indicators were more favorable than that of N fertilizer. PMID- 12549539 TI - Stable sulfur isotope ratio indicates long-term changes in sulfur deposition in the Broadbalk experiment since 1845. AB - Archived wheat (Triticum aestivum L.) grain and straw, and soil samples from the control plot of the Rothamsted Broadbalk Experiment, located in southeastern England and established in 1843, were used to investigate the effects of dramatically changing SO2 pollution inputs on the concentrations and stable isotope ratios (delta34S) of S in the samples. Representative coal samples from UK major coal fields were also determined for delta34S. Concentrations of S showed no clear trends in either grain or straw over the 155 years from 1845 to 1999. However, grain and straw delta34S decreased rapidly from 6 to 7/1000 in 1845 to -2 to -5/1000 in the early 1970s, and since then have increased to 0.5 to 2/1000 in the late 1990s. This pattern mirrored the trend of UK SO2 emissions over the 155 years. Both grain and straw delta34S correlated strongly and negatively with UK SO2 emissions (R2 > 0.89), but the relationships were different for the pre- and post-1970 data sets. Soil delta34S also decreased considerably, from 8.2/1000 in 1865 to 3.7 to 4.5/1000 during 1965-1999. A negative delta34S value was inferred for the anthropogenic S deposited at the experimental site before 1970, and further confirmed by negative delta34S values (-6 to -10/1000) found in the coal samples from southeastern England and southern Wales. Based on the S isotope ratios, we estimated that anthropogenic S contributed 62 to 78% of the S uptake by wheat at the peak of SO2 emissions, and accounted for 28 to 37% of the topsoil S in 1965. PMID- 12549540 TI - Transport and biodegradation of perchlorate in soils. AB - Perchlorate (ClO4-) contamination of ground water and surface water is a widespread problem, particularly in the western United States. This study examined the effect of biodegradation on perchlorate fate and transport in soils. Solute transport experiments were conducted on two surface soils. Pulses of solution containing perchlorate and Br- were applied to saturated soil columns at steady state water flow. Perchlorate behaved like a nonreactive tracer in Columbia loam (coarse-loamy, mixed, superactive, nonacid, thermic Oxyaquic Xerofluvent) but was degraded in Yolo loam (fine-silty, mixed, superactive, nonacid, thermic Mollic Xerofluvent). Batch experiments demonstrated that perchlorate removal from solution in Yolo loam was caused by biodegradation. Other batch experiments with Yolo loam surface and subsurface soils, Columbia loam surface soil, and dredge tailings demonstrated that perchlorate biodegradation required anaerobic conditions, an adequate carbon source, and an active perchlorate-degrading microbial population. The sequential reduction of perchlorate and NO3- by an indigenous soil microbial community in Yolo loam batch systems was also studied. Nitrate reduction occurred much sooner than perchlorate reduction in soils that had not been previously exposed to perchlorate, but NO3- and perchlorate were simultaneously reduced in soils previously exposed to perchlorate. The results of this study have implications for in situ remediation schemes and for agricultural soils that have been contaminated by perchlorate tainted irrigation water. PMID- 12549541 TI - Enrichment and isolation of endosulfan-degrading microorganisms. AB - Endosulfan (6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,3,4 benzo-dioxathiepin-3-oxide) is a cyclodiene organochlorine currently used as an insecticide all over the world and its residues are posing a serious environmental threat. This study reports the isolation and identification of enriched microorganisms, capable of degrading endosulfan. Enrichment was achieved by using the insecticide as either the sole source of carbon or sulfur in parallel studies. Two strains each of fungi (F1 and F4) and bacteria (BF2 and B4) were selected using endosulfan as a sole carbon source. A Pandoraea species (Lin 3) previously isolated in our laboratory using lindane (gamma-HCH) as a carbon source was also screened for endosulfan degradation. F1 and F4 (Fusarium ventricosum) degraded alpha-endosulfan by as much as 82.2 and 91.1% and beta endosulfan by 78.5 and 89.9%, respectively, within 15 d of incubation. Bacterial strains B4 and Lin-3 degraded alpha-endosulfan up to 79.6 and 81.8% and beta endosulfan up to 83.9 and 86.8%, respectively, in 15 d. Among the bacterial strains isolated by providing endosulfan as a sulfur source, B4s and F4t degraded alpha-endosulfan by as much as 70.4 and 68.5% and beta-endosulfan by 70.4 and 70.8%, respectively, after 15 d. Degradation of the insecticide occurred concomitant with bacterial growth reaching an optical density (OD600) of 0.366 and 0.322 for B4 and Lin-3, respectively. High OD600 was also noted with the other bacterial strains utilizing endosulfan as a sulfur source. Fungal and bacterial strains significantly decreased the pH of the nutrient culture media while growing on endosulfan. The results of this study suggest that these novel strains are a valuable source of potent endosulfan-degrading enzymes for use in enzymatic bioremediation. PMID- 12549542 TI - The potential of rhizosphere microbes isolated from a constructed wetland to biomethylate selenium. AB - The potential of rhizosphere microbes isolated from common reed [Phragmites australis (Cav.) Trin. ex Steud] plants grown in a subsurface-flow constructed wetland to biomethylate selenate or selenite was studied in liquid cultures under controlled conditions. Total mean percentages of volatilized Se from half strength Hoagland culture solutions (low C content) supplemented with selenate or selenite and inoculated with cultured rhizosphere microbes after 15 d of incubation were 7.9 and 49.1%, respectively. There was a relative best fit (r = 0.87) between total number of rhizosphere and cultured microbes and the percentage of volatilized Se in Hoagland solution after 15 d of incubation. However, when the same microbes were cultured in tryptic soybean broth (TSB) medium (high C content), the percentages of volatilized Se from selenate and selenite were 1.3 and 1.9%, respectively. The volatilization percentages of Se from selenate or selenite in culture solutions inoculated with rhizosphere suspension instead of cultured rhizosphere microbes were very low (1.2-3.0%) in both cultivation media. In all experiments, selenite was volatilized significantly (p < 0.05) in higher amounts by cultured rhizosphere microbes after 15 d of incubation compared with selenate. Dissolved biomethylated dimethylselenide (DMSe) in water samples taken from the subsurface-flow bed was determined by purging with helium. The DMSe in water samples was indirectly detected up to 2.4 microg Se L(-1), which indicates that part of the produced DMSe was dissolved in the matrix before being released into the atmosphere. Our results show that rhizosphere microbes isolated from common reed plants have a high potential of Se biomethylation and volatilization from selenate and selenite. PMID- 12549543 TI - Enzymatic oxidative transformation of chlorophenol mixtures. AB - Chlorinated phenols are major industrial and agricultural xenobiotics that pollute soil and ground water. It has been shown that laccases catalyze the oxidative coupling of phenolic compounds. Therefore, the transformation of one or a mixture of several chlorinated phenols by a laccase from the fungus Trametes villosa was studied. Generally, if more than one phenol was added, the transformation of chlorinated phenols decreased, and if the concentration of the laccase was increased, the transformation of the phenols was enhanced. There were exceptions to these observations: for instance, the transformation of 0.1 mM 4 chlorophenol incubated with 1 mM 2,4-dichlorophenol in buffered salt solutions was not enhanced if the concentration of the laccase was increased from 2 to 20 DMP units/mL. The reason for the reduced transformation of chlorinated phenols in the presence of additional phenols is still unknown. However, in spite of some limitations, the application of laccase to decontaminate wastewater polluted with chlorinated phenols appears feasible. PMID- 12549544 TI - Determination of redox-sensitive phosphorus in field runoff without sediment preconcentration. AB - Reduction-induced phosphorus (P) release from particles transported by field runoff has been poorly studied for want of a method that could be used for large surveys. To rectify this shortcoming, we modified the bicarbonate-dithionite (BD) extraction step of a sediment P speciation scheme for analyzing redox-sensitive P in runoff without sample preconcentration. The extraction comprised the addition of bicarbonate (pH buffer) and dithionite (reducing agent) into a runoff sample, 15 min of gentle shaking, filtration, and sample digestion. The samples were greatly reduced (Eh < -200 mV), and Fe and P were solubilized, but Al solubility was not increased. Phosphorus release from rock phosphates (calcium phosphates) was greater in the BD extraction than in water or bicarbonate solution, although no more than 0.2% of the total P was released. For runoff from a very fine Typic Cryaquept, the particulate phosphorus (PP) versus BD-PP relationship was linear up to a PP concentration of about 1.0 mg L(-1), but over the whole PP range studied (up to 2.6 mg L(-1)) somewhat better described by an exponential equation (BD-PP = 0.297 x PP(0.766); r2 = 0.91, n = 79). The minimum detectable value given by the method was relatively low, 0.023 mg L(-1), but reproducibility varied, with the coefficient of variation for 10 samples analyzed with 5 replicates ranging from 1.8 to 28.5%. Considering the variable reproducibility of the results and the lack of suitable reference material, the method needs further refinement and testing if it is to be used for quantitative determination of redox-sensitive P in runoff. PMID- 12549545 TI - Simazine runoff from citrus orchards affected by shallow mechanical incorporation. AB - Simazine (6-chloro-N,N'-diethyl-1,3,5-triazine-2,4-diamine) losses via runoff in California are a potential source of environmental contamination because simazine is widely used for weed control during the rainy season from November to March. This study was conducted in two citrus orchards from three rainfall events to evaluate the effects of shallow mechanical incorporation on simazine losses in runoff during the winter. Simazine losses in runoff were compared between row middles that were either undisturbed, the normal orchard practice, or subjected to shallow mechanical incorporation. Mechanical incorporation of row middles significantly reduced runoff volumes by approximately 45 and 28% for the first and second runoff events, respectively. In undisturbed plots, simazine concentrations in runoff from the first runoff event ranged from 0.62 to 0.73 mg L(-1); then simazine concentrations rapidly decreased (0.03-0.35 mg L(-1)) from the second and third runoff events. In disturbed plots, simazine concentrations in runoff from the first runoff event ranged from 0.21 to 0.24 mg(-1), but simazine concentrations remained relatively constant between the three runoff events. Total mass recoveries of simazine in runoff ranged from 1.93 to 2.97% and from 0.70 to 0.74% of application from the undisturbed plots and from the disturbed plots, respectively. Low water infiltration rate inhibited surface applied herbicide incorporation into the soil matrix with natural rainfall in compacted soils. Mechanical incorporation of row middles significantly reduced runoff volumes, simazine concentrations, and mass losses in runoff after application. PMID- 12549546 TI - Herbicide loading to shallow ground water beneath Nebraska's Management Systems Evaluation Area. AB - Better management practices can counter deterioration of ground water quality. From 1991 through 1996 the influence of improved irrigation practices on ground water pesticide contamination was assessed at the Nebraska Management Systems Evaluation Area. Three 13.4-ha corn (Zea mays L.) fields were studied: a conventional furrow-irrigated field, a surge-irrigated field and a center pivot irrigated field, and a center pivot-irrigated alfalfa (Medicago sativa L.) field. The corn fields received one identical banded application of Bicep (atrazine [6 chloro-N-ethyl-N'-(1-methylethyl)-1,3,5-triazine-2,4,-diamine] + metolachlor [2 chloro-N-(2-ethyl-6-methylphenyl)-N-(2-methoxy-1-methylethyl) acetamidel) annually; the alfalfa field was untreated. Ground water samples were collected three times annually from 16 depths of 31 multilevel samplers. Six years of sample data indicated that a greater than 50% reduction in irrigation water on the corn management fields lowered average atrazine concentrations in the upper 1.5 m of the aquifer downgradient of the corn fields from approximately 5.5 to <0.5 microg L(-1). Increases in deethylatrazine (DEA; 2-chloro-4-amino-6 isopropylamino-s-triazine) to atrazine molar ratios indicated that reducing water applications enhanced microbial degradation of atrazine in soil zones. The occurrence of peak herbicide loading in ground water was unpredictable but usually was associated with heavy precipitation within days of herbicide application. Focused recharge of storm runoff that ponded in the surge-irrigated field drainage ditch, in the upgradient road ditch, and at the downgradient end of the conventionally irrigated field was a major mechanism for vertical transport. Sprinkler irrigation technology limited areas for focused recharge and promoted significantly more soil microbial degradation of atrazine than furrow irrigation techniques and, thereby, improved ground water quality. PMID- 12549547 TI - Herbicides in ground water beneath Nebraska's Management Systems Evaluation Area. AB - Profiles of ground water pesticide concentrations beneath the Nebraska Management Systems Evaluation Area (MSEA) describe the effect of 20 yr of pesticide usage on ground water in the central Platte Valley of Nebraska. During the 6-yr (1991 1996) study, 14 pesticides and their transformation products were detected in 7848 ground water samples from the unconfined water table aquifer. Triazine and acetamide herbicides applied on the site and their transformation products had the highest frequencies of detection. Atrazine [6-chloro-N-ethyl-N'-(1 methylethyl)-1,3,5-triazine-2,4,-diamine] concentrations decreased with depth and ground water age determined with 3H/3He dating techniques. Assuming equivalent atrazine input during the past 20 yr, the measured average changes in concentration with depth (age) suggest an estimated half-life of >10 yr. Hydrolysis of atrazine and deethylatrazine (DEA; 2-chloro-4-amino-6 isopropylamino-s-triazine) to hydroxyatrazine [6-hydroxy-N-ethyl-N'-(1 methylethyl)-1,3,5-triazine-2,4-diamine] appeared to be the major degradation route. Aqueous hydroxyatrazine concentrations are governed by sorption on the saturated sediments. Atrazine was detected in the confined Ogallala aquifer in ultra-trace concentrations (0.003 microg L(-1)); however, the possibility of introduction during reverse circulation drilling of these deep wells cannot be eliminated. In fall 1997 sampling, metolachlor [2-chloro-N-(2-ethyl-6 methylphenyl)-N-(2-methoxy-1-methylethyl) acetamide] was detected in 57% of the 230 samples. Metolachlor oxanilic acid [(2-ethyl-6-methylphenyl)(2-methoxy-1 methylethyl) amino]oxo-acetic acid] was detected in most samples. In ground water profiles, concentrations of metolachlor ethane sulfonic acid [2-[(ethyl-6 methylphenyl)(2-methoxy-1-methylethyl)amino]-2-oxo-ethanesulfonic acid] exceeded those of deethylatrazine. Alachlor [2-chloro-N-(2,6-diethylphenyl)-N (methoxymethyl)acetamide] was detected in <1% of the samples; however, alachlor ethane sulfonic acid [2-[(2,6-diethylphenyl)(methoxymethyl)amino]-2 oxoethanesulfonic acid] was present in most samples (63%) and was an indicator of past alachlor use. PMID- 12549548 TI - Effect of biosolids processing on lead bioavailability in an urban soil. AB - The potential for biosolids products to reduce Pb availability in soil was tested on a high Pb urban soil with biosolids from a treatment plant that used different processing technologies. High Fe biosolids compost and high Fe + lime biosolids compost from other treatment plants were also tested. Amendments were added to a Pb-contaminated soil (2000 mg kg(-1) Pb) at 100 g kg(-1) soil and incubated for 30 d. Reductions in Pb bioavailability were evaluated with both in vivo and in vitro procedures. The in vivo study entailed feeding a mixture of the Pb contaminated soil and AIN93G Basal Mix to weanling rats. Three variations of an in vitro procedure were performed as well as conventional soil extracts [diethylenetriaminepentaacetic acid (DTPA) and Ca(NO3)2] and sequential extraction. Addition of the high Fe compost reduced the bioavailability of soil Pb (in both in vivo and in vitro studies) by 37 and 43%, respectively. Three of the four compost materials tested reduced Pb bioavailability more than 20%. The rapid in vitro (pH 2.3) data had the best correlation with the in vivo bone results (R = 0.9). In the sequential extract, changes in partitioning of Pb to Fe and Mn oxide fractions appeared to reflect the changes in in vivo Pb bioavailability. Conventional extracts showed no changes in metal availability. These results indicate that addition of 100 g kg(-1) of high Fe and Mn biosolids composts effectively reduced Pb availability in a high Pb urban soil. PMID- 12549549 TI - Arsenic distribution in Florida urban soils: comparison between Gainesville and Miami. AB - Arsenic contamination is of concern due to its effect as a carcinogen. Understanding the distribution of arsenic in urban soils is important for establishing baseline concentrations from which anthropogenic effects can be measured. The soil cleanup target level (SCTL) for arsenic in Florida (0.8 and 3.7 mg kg(-1) in residential and commercial areas, respectively) is lower than in most states and is near the arsenic background concentrations in Florida soils. The objective of this study was to characterize the distribution of arsenic in the soils of two Florida cities, Gainesville and Miami. More than 200 soil samples were collected from three land-use classes in each city (residential, commercial, and public land), digested with USEPA Method 3051a, and analyzed with graphite furnace atomic absorption spectrophotometry. Arsenic concentrations varied greatly in Gainesville, ranging from 0.21 to approximately 660 mg kg(-1) with a geometric mean (GM) of 0.40 mg kg(-1) (after discarding outliers), which was significantly lower than the GM of 2.81 mg kg(-1) in Miami, although Miami samples ranged only from 0.32 to approximately 110 mg kg(-1). Arsenic concentrations in 29 and 4% of the Gainesville soil samples and 95 and 33% of the Miami samples exceeded the Florida residential and commercial SCTL, respectively. This study is the first to provide information on arsenic distribution in urban soils of Florida, and the data are useful for assessing arsenic contamination and determining the need for remediation. PMID- 12549550 TI - Effects of organic amendments on the reduction and phytoavailability of chromate in mineral soil. AB - In this study, seven organic amendments (biosolid compost, farm yard manure, fish manure, horse manure, spent mushroom, pig manure, and poultry manure) were investigated for their effects on the reduction of hexavalent chromium [chromate, Cr(VI)] in a mineral soil (Manawatu sandy soil) low in organic matter content. Addition of organic amendments enhanced the rate of reduction of Cr(VI) to Cr(III) in the soil. At the same level of total organic carbon addition, there was a significant difference in the extent of Cr(VI) reduction among the soils treated with organic amendments. There was, however, a significant positive linear relationship between the extent of Cr(VI) reduction and the amount of dissolved organic carbon in the soil. The effect of biosolid compost on the uptake of Cr(VI) from the soil, treated with various levels of Cr(VI) (0-1200 mg Cr kg(-1) soil), was examined with mustard (Brassica juncea L.) plants. Increasing addition of Cr(VI) increased Cr concentration in plants, resulting in decreased plant growth (i.e., phytotoxicity). Addition of the biosolid compost was effective in reducing the phytotoxicity of Cr(VI). The redistribution of Cr(VI) in various soil components was evaluated by a sequential fractionation scheme. In the unamended soil, the concentration of Cr was higher in the organic bound, oxide-bound, and residual fractions than in the soluble and exchangeable fractions. Addition of organic amendments also decreased the concentration of the soluble and exchangeable fractions but especially increased the organic-bound fraction in soil. PMID- 12549551 TI - Influence of soil geochemical and physical properties on the sorption and bioaccessibility of chromium(III). AB - There are numerous Cr(III)-contaminated sites on Department of Defense (DoD) and Department of Energy (DOE) lands that are awaiting possible clean up and closure. Ingestion of contaminated soil by children is the risk driver that generally motivates the likelihood of site remediation. The purpose of this study was to develop a simple statistical model based on common soil properties to estimate the hioaccessibility of Cr(III)-contaminated soil upon ingestion. Thirty-five uncontaminated soils from seven major soil orders, whose properties were similar to numerous U.S. DoD contaminated sites, were treated with Cr(III) and aged. Statistical analysis revealed that Cr(III) sorption (e.g., adsorption and surface precipitation) by the soils was strongly correlated with the clay content, total inorganic C, pH, and the cation exchange capacity of the soils. Soils with higher quantities of clay, inorganic C (i.e., carbonates), higher pH, and higher cation exchange capacity generally sequestered more Cr(III). The amount of Cr(III) bioaccessible from the treated soils was determined with a physiologically based extraction test (PBET) that was designed to simulate the digestive process of the stomach. The bioaccessibility of Cr(III) varied widely as a function of soil type with most soils limiting bioaccessibility to <45 and <30% after I and 100 d soil Cr aging, respectively. Statistical analysis showed the bioaccessibility of Cr(III) on soil was again related to the clay and total inorganic carbon (TIC) content of the soil. Bioaccessibility decreased as the soil TIC content increased and as the clay content decreased. The model yielded an equation based on common soil properties that could be used to predict the Cr(III) bioaccessibility in soils with a reasonable level of confidence. PMID- 12549552 TI - Correlation of cadmium distribution coefficients to soil characteristics. AB - Cadmium (Cd) distribution between the soil solid phase and the soil solution is a key issue in assessing the environmental effect of Cd in the terrestrial environmental. Previous studies have shown that many individual minerals and other components found in soils can bind Cd, but most studies on whole soil samples have shown that pH is the main parameter controlling the distribution. To identify further the components that are important for Cd binding in soil we measured Cd distribution coefficients (Kd) at two fixed pH values and at low Cd loadings for 49 soils sampled in Denmark. The Kd values for Cd ranged from 5 to 3000 L kg(-1). The soils were described pedologically and characterized in detail (22 parameters) including determination of contents of the various minerals in the clay fraction. Correlating parameters were grouped and step-wise regression analysis revealed that the organic carbon content was a significant variable at both pH values. Cation exchange capacity (CEC) and gibbsite were important at the low pH (5.3) while iron oxides also were important at the high pH (6.7). None of the other clay minerals present in the soils (illite, smectite, kaolinite, hydroxy interlayered clay minerals [HIM], chlorite, quartz, microcline, plagioclase) were significant in explaining the Cd distribution coefficient. PMID- 12549553 TI - Long-term biosolids application effects on metal concentrations in soil and bermudagrass forage. AB - The long-term application of biosolids that periodically contained elevated metal concentrations has raised questions about potential effects on animal health. To address these concerns, we determined metal concentrations (As, Cd, Cu, Pb, Hg, Mo, Ni, Se, and Zn) in both soil and bermudagrass [Cynodon dactylon (L.) Pers.] forage from 10 fields in the following categories of biosolids application: six or more years (>6YR), less than six years (<6YR), and no applications (NS). Soil metal concentrations in all groups were similar to values reported for mineral soils in Georgia, and well below USEPA cumulative limits. Average metal concentrations in the forage were below the maximum tolerable level (MTL) for beef cattle, although two biosolids-amended fields in the >6YR group produced forage that was at or near the MTL for Cd and Mo, and one field in the <6YR group produced forage above the MTL for Cd. The Cu to Mo ratios in forage decreased with increasing time of sludge application, with the average in the >6YR group at a proposed 5:1 Cu to Mo ratio limit to protect ruminant health. Sulfur concentrations in the forage from all three groups was near the MTL of 4 g kg( 1). The study indicated that toxic levels of metals have not accumulated in the soils due to long-term biosolids application. Overall forage quality from the biosolids-amended fields was similar to that of commercially fertilized fields; however, due to the relatively high S and potential for a low Cu to Mo ratio, Cu supplements should be used to ensure ruminant health. PMID- 12549554 TI - In situ treatment of metals in contaminated soils with phytate. AB - Batch experiments were conducted to evaluate the ability of various forms of phytate, the hexaphosphoric form of myo-inositol (IP6), to immobilize U, Ni, and other inorganic contaminants in soils and sediments. A Ca-phytate precipitate (Ca(n)-IP6), dodeca sodium-phytate (Na12-IP6), and hydroxyapatite (HA) were added to contaminated soil at rates of 0, 10, 25, and 50 g kg(-1) and equilibrated in 0.001 M CaCl2. The samples were then centrifuged, the solution pH was measured, and the supernatants were filtered prior to analysis for dissolved organic carbon (DOC), U, Ni, P, and other inorganic contaminants, such as As, Cr, Se, and Pb. The residual sediments were air-dried prior to characterization by analytical electron microscopy and extraction with the Toxicity Characteristic Leaching Procedure (TCLP). The solubility of several metals (e.g., U, Pb, Cu) increased with increasing Na12-IP6 when compared with the nonamended control. In some cases immobilization was observed at the lowest Na12-IP6 application rate (10 g kg(-1)) with an increase in solubility observed at the higher rates, demonstrating the importance of metal to ligand ratio. In contrast, Ca(n)-IP6 and HA decreased the solubility of U, Ni, Al, Pb, Ba, Co, Mn, and Zn. For example, soluble U decreased from 2242 to 76 microg kg(-1) and Ni from 58 to 9.6 mg kg with the Ca(n)-IP6 addition, similar to the results observed for HA. Arsenic and Se solubility increased for HA and both forms of IP6, but to a much greater degree for Na12 IP6, suggesting that the increase in pH observed for HA and Na12-IP6, combined with added competition from PO4 and IP6 for sorption sites, resulted in the release of sorbed oxyanion contaminants. The analytical electron microscopy results indicated that metals such as U and Ni were closely associated with secondary Al-rich precipitates in the HA-treated soils, rather than unreacted HA. The analytical electron microscopy results were less definitive for the Ca(n)-IP6 treated soil, although the residual P-containing material was enriched in Al, with lesser amounts of U and Ni. PMID- 12549555 TI - Nitrate removal effectiveness of a riparian buffer along a small agricultural stream in western Oregon. AB - The Willamette Valley of Oregon has extensive areas of poorly drained, commercial grass seed lands. Little is know about the ability of riparian areas in these settings to reduce nitrate in water draining from grass seed fields. We established two study sites with similar soils and hydrology but contrasting riparian vegetation along an intermittent stream that drains perennial ryegrass (Lolium perenne L.) fields in the Willamette Valley of western Oregon. We installed a series of nested piezometers along three transects at each site to examine NO3-N in shallow ground water in grass seed fields and riparian areas. Results showed that a noncultivated riparian zone comprised of grasses and herbaceous vegetation significantly reduced NO3-N concentrations of shallow ground water moving from grass seed fields. Darcy's law-based estimates of shallow ground water flow through riparian zone A/E horizons revealed that this water flowpath could account for only a very small percentage of the streamflow. Even though there is great potential for NO3-N to be reduced as water moves through the noncultivated riparian zone with grass-herbaceous vegetation, the potential was not fully realized because only a small proportion of the stream flow interacts with riparian zone soils. Consequently, effective NO3-N water quality management in poorly drained landscapes similar to the study watershed is primarily dependent on implementation of sound agricultural practices within grass seed fields and is less influenced by riparian zone vegetation. Wise fertilizer application rates and timing are key management tools to reduce export of NO3-N in stream waters. PMID- 12549556 TI - Diuron occurrence and distribution in soil and surface and ground water associated with grass seed production. AB - Little is known about the occurrence and distribution of the herbicide diuron [3 (3,4-dichlorophenyl)-1,1-dimethyl urea] in soil, ground water, and surface water in areas affected by grass-seed production. A field study was designed to investigate the occurrence and distribution of diuron and its transformation products at a poorly drained field site located along an intermittent tributary of Lake Creek in the southern Willamette Valley of Oregon. The experimental sites consisted of a field under commercial grass seed production with a cultivated riparian zone and a second site that was part of the same grass seed field but with a noncultivated riparian zone. Diuron and its transformation product DCPMU [3-(3,4-dichlorophenyl)-1-methylurea] were the only significant residues detected in this study. Concentrations of diuron in surface water declined from a maximum of 28 microg/L immediately following application to low levels that persisted as long as flow was present. Diuron and DCPMU concentrations in shallow ground water (15-36 cm below ground surface) were highest (2-13 microg/L) in the zone immediately adjacent (0.5 m) to Lake Creek and indicated the influence of stream water on shallow ground water near the stream. Diuron and DCPMU detected in soil prior to the second season's application indicated the persistence of diuron and DCPMU from the previous year's application. Surface runoff during the rainy season removes only a very small percentage (<1%) of the applied herbicide. In addition, no evidence was obtained for the downward transport of diuron or its transformation products to deep ground water. PMID- 12549557 TI - Identifying relationships between baseflow geochemistry and land use with synoptic sampling and R-mode factor analysis. AB - The relationship between land use and stream chemistry is often explored through synoptic sampling of rivers at baseflow conditions. However, baseflow chemistry is likely to vary temporally and spatially with land use. The purpose of our study is to examine the usefulness of the synoptic sampling approach for identifying the relationship between complex land use configurations and stream water quality. This study compares biogeochemical data from three synoptic sampling events representing the temporal variability of baseflow chemistry and land use using R-mode factor analysis. Separate R-mode factor analyses of the data from individual sampling events yielded only two consistent factors. Agricultural activity was associated with elevated levels of Ca2+, Mg2+, alkalinity, and frequently K+, SO4(2-), and NO3-. Urban areas were associated with higher concentrations of Na+, K+, and Cl-. Other retained factors were not consistent among sampling events, and some factors were difficult to interpret in the context of biogeochemical sources and processes. When all data were combined, further associations were revealed such as an inverse relationship between the proportion of wetlands and stream nitrate concentrations. We also found that barren lands were associated with elevated sulfate levels. This research suggests that an individual sampling event is unlikely to characterize adequately the complex processes controlling interactions between land use and stream chemistry. Combining data collected over two years during three synoptic sampling events appears to enhance our ability to understand processes linking stream chemistry and land use. PMID- 12549558 TI - Leaf litter dynamics and nitrous oxide emission in a Mediterranean riparian forest: implications for soil nitrogen dynamics. AB - Mediterranean riparian zones can experience severe drought periods that lead to low soil moisture content, which dramatically affects their performance as nitrate removal systems. In the Mediterranean riparian zone of this study, we determined that N2O emission was practically nil. To understand the role of forest floor processes in nitrogen retention of a Mediterranean riparian area, we studied leaf litter dynamics of two tree species, London planetree [Platanus x acerifolia (Aiton) Willd.] and alder [Alnus glutinosa (L.) Gaertn.], for two years, along with soil nitrogen mineralization rates. Annual leaf litter fall equaled 562.6 +/- 10.1 (standard error) g dry wt. m(-2), 68% of which was planetree and 32% of which was alder. The temporal distribution of litterfall showed a two-peak annual cycle, one occurring in midsummer, the other in autumn. Planetree provided the major input of organic nitrogen to the forest floor, and the amount of planetree leaves remaining on the forest floor was equivalent to approximately four years of stock. Leaf litter decomposition was three times higher for alder (decay coefficient [k] = 1.13 yr(-1)) than for planetree (k = 0.365 yr(-1)). Mineralization rates showed a seasonal pattern, with the maximum rate in summer (1.92 mg N kg(-1) d(-1)). Although the forest floor was an important sink for nitrogen due to planetree leaf accumulation, 7.5% of this leaf litter was scoured to the streambed by wind. This loss was irrelevant for alder leaves. Due to the litter quality, the forest floor of this Mediterranean riparian forest acts as a nitrogen sink. PMID- 12549559 TI - Stable carbon isotope ratio and composition of microbial fatty acids in tropical soils. AB - The soil microbial community plays a critical part in tropical ecosystem functioning through its role in the soil organic matter (SOM) cycle. This study evaluates the relative effects of soil type and land use on (i) soil microbial community structure and (ii) the contribution of SOM derived from the original forest vegetation to the functioning of pasture and sugarcane (Saccharum spp.) ecosystems. We used principal components analysis (PCA) of soil phospholipid fatty acid (PLFA) profiles to evaluate microbial community structure and PLFA stable carbon isotope ratios (delta13C) as indicators of the delta13C of microbial substrates. Soil type mainly determined the relative proportions of gram positive versus gram negative bacteria whereas land use primarily determined the relative proportion of fungi, protozoa, and actinomycetes versus other types of microorganisms. Comparison of a simple model to our PLFA delta13C data from land use chronosequences indicates that forest-derived SOM is actively cycled for appreciably longer times in sugarcane ecosystems developed on Andisols (mean turnover time = 50 yr) than in sugarcane ecosystems developed on an Oxisol (mean turnover time = 13 yr). Our analyses indicate that soil chronosequence PLFA delta13C measurements can be useful indicators of the contribution that SOM derived from the original vegetation makes to continued ecosystem function under the new land use. PMID- 12549560 TI - The effects of soil carbon on phosphorus and sediment loss from soil trays by overland flow. AB - Soil chemical constituents influence soil structure and erosion potential. We investigated manure and inorganic fertilizer applications on soil chemistry (carbon [C] quality and exchangeable cations), aggregation, and phosphorus (P) loss in overland flow. Surface samples (0-5 cm) of a Hagerstown (fine, mixed, semiactive, mesic Typic Hapludalf) soil, to which either dairy or poultry manure or triple superphosphate had been applied (0-200 kg P ha(-1) yr(-1) for 5 yr), were packed in boxes (1 m long, 0.15 m wide, and 0.10 m deep) to field bulk density (1.2 g cm(-3)). Rainfall was applied (65 mm h(-1)), overland flow collected, and sediment and P loss determined. All amendments increased Mehlich 3 extractable P (19-177 mg kg(-1)) and exchangeable Ca (4.2-11.5 cmol kg(-1)) compared with untreated soil. For all treatments, sediment transport was inversely related to the degree of soil aggregation (determined as ratio of dispersed and undispersed clay; r = 0.51), exchangeable Ca (r = 0.59), and hydrolyzable carbohydrate (r = 0.62). The loss of particulate P and total P in overland flow from soil treated with up to 50 kg P ha(-1) dairy manure (9.9 mg particulate phosphorus [PPI, 15.1 mg total phosphorus [TP]) was lower than untreated soil (13.3 mg PP, 18.1 mg TP), due to increased aggregation and decreased surface soil slaking attributed to added C in manure. Manure application at low rates (<50 kg P ha(-1)) imparts physical benefits to surface soil, which decrease P loss potential. However, at greater application rates, P transport is appreciably greater (26.9 mg PP, 29.5 mg TP) than from untreated soil (13.3 mg PP, 18.1 mg TP). PMID- 12549561 TI - Turfgrass thatch effects on pesticide leaching: a laboratory and modeling study. AB - Process-based models are frequently used to assess the water quality impacts of turfgrass management emanating from proposed or existing golf courses. Thatch complicates the prediction of pesticide transport because surface-applied pesticides must pass through an organic-rich layer before entering the soil. This study was conducted to (i) compare the use of a linear equilibrium model (LEM) and two-site nonequilibrium (2SNE) model to predict pesticide transport through soil and thatch + soil columns, and (ii) evaluate thatch effects on pesticide transport through soil columns with a volume-averaging approach. Pesticide breakthrough curves were obtained for soil and thatch + soil columns from a 1 cm h(-1) flux applied one day after applying triclopyr (3,5,6-trichloro-2 pyridinyloxyacetic acid) and carbaryl (1-napthyl-methyl carbamate). Pesticide and bromide transport parameters indicated that nonequilibrium processes were affecting pesticide transport. Columns containing zoysiagrass (Zoysia japonica Steud.) thatch had lower triclopyr and carbaryl leaching losses than did soil only columns, although total reductions attributable to thatch did not exceed 15% of the applied pesticide. When laboratory-based retardation factors were used, the 2SNE model explained 88 to 93% of the variability for triclopyr and 70 to 94% of the variability for carbaryl. Laboratory-based retardation factors performed well in a 2SNE model to predict the peak concentration and tailing behavior of triclopyr and carbaryl with a volume-averaging approach. These results suggest that separate representation of the thatch layer in process-based models is not a prerequisite to obtain reasonable estimates of pesticide transport under steady state flow conditions. PMID- 12549562 TI - Persistent organic pollutants in the environment: chlordane residues in compost. AB - The half-lives of some persistent organic pollutants (POPs) in environmental compartments such as soil and air can be as long as decades. In spite of the hydrophobicity of many POPs, the literature contains reports of their uptake by, and translocation through, a variety of plants. Both these observations prompt the investigation of whether a vegetation-based environmental compartment such as compost contains significant residues of POPs. Previous reports imply that residues of technical chlordane will be found in compost. Due to its physicochemical properties, technical chlordane provides insights into the fate of POPs in the environment, which are not accessible through determinations of other pollutants in this group. Accordingly, we undertook the first comprehensive examination of technical chlordane residues in a variety of composts, specifically, 13 commercial and 39 municipal compost products, to both characterize and quantify the magnitude of this point source of contamination. Using chiral gas chromatography interfaced to ion trap mass spectrometry, the concentration and the compositional and enantiomeric profiles of chlordane components were determined. Of the 13 commercial products, 9 contained detectable chlordane concentrations, ranging from 4.7 to 292 microg/kg (dry wt.), while all 39 municipal products contained chlordane residues ranging from 13.9 to 415 microg/kg (dry wt.). The residue concentrations and profiles suggest possible feedstock sources for the chlordane in the finished compost product. The data also support the conclusion that some composts contribute to anthropogenic cycling of POPs through the biosphere. PMID- 12549563 TI - Molecular-level investigation of monoaromatic compound sorption to suspended soil particles by deuterium nuclear magnetic resonance. AB - Molecular-level sorption behavior of monoaromatic compounds in suspensions of water-dispersable clay components was studied by measuring 2H nuclear magnetic resonance (NMR) spin-spin relaxation times (T2). In general, decreased T2 values indicate stronger solute-sorbent interactions and increased sorption of the solute. A decreasing trend for T2 values in the order benzene > fluorobenzene > toluene (-C6D5 moiety) was observed, which was probably caused by the hydrophobic effect. The T2 values for benzene and the -C6D5 moiety of toluene increased with increasing pH, whereas the trend with pH was much weaker and less consistent for fluorobenzene and the methyl group of toluene. Conversely, no clear relationship was found between T2 values and pH for dichloromethane. These contrasting results cannot be explained by the pH-dependent self-assembly and hydrophobicity of humics. Instead, directed specific forces, including hydrogen bonding, cation-pi interactions, and aromatic-aromatic interactions, are proposed between the benzene ring of monoaromatic solutes and soil organic matter (SOM). Substituents of benzene affect these interactions by varying the pi electron density. When the soil fraction was treated with NaOH to remove humic and fulvic acids, T2 values for the different monoaromatic solutes were surprisingly lower compared with those for the untreated soil fraction. This result is probably caused by the increased ratio of solutes adsorbed to "hard" or "glassy" SOM components, which leads to less mobile sorbed solute molecules, after removing NaOH-extractable humics that contain more "soft" or "rubbery" SOM components. PMID- 12549564 TI - Sorption and desorption of naphthalene by soil organic matter: importance of aromatic and aliphatic components. AB - Nonlinear isotherm behavior has been reported for the sorption of hydrophobic organic compounds (HOCs) in soil organic matter (SOM), but the exact mechanisms are unknown. Our objective was to provide insight into the sorption mechanism of HOCs in SOM by studying the sorption-desorption processes of naphthalene in a mineral soil, its humic fractions, and lignin. Additionally, humin and lignin were used for studying the effects of temperature and cosolvent on HOC sorption. All isotherms were nonlinear. The humin and lignin isotherms became more linear at elevated temperatures and with the addition of methanol indicating a condensed to expanded structural phase transition. Isotherm nonlinearity and hysteresis increased in the following order: soil humic acid (HA) < soil < soil humin. Of the samples, aliphatic-rich humin exhibited the largest degree of nonlinearity and had the highest sorption capacity for naphthalene. High nonlinearity and hysteresis in humin were most likely caused by its condensed structure. A novel aliphatic, amorphous condensed conformation is proposed. This conformation can account for both high sorption capacities and increased nonlinearity observed for aliphatic-rich samples and can explain many sorption disparities discussed in the literature. This study clearly illustrates the importance of both aliphatic and aromatic moieties for HOC sorption in SOM. PMID- 12549565 TI - Integrated BIosphere Simulator (IBIS) yield and nitrate loss predictions for Wisconsin maize receiving varied amounts of nitrogen fertilizer. AB - Agriculture in the U.S. Midwest faces the formidable challenge of improving crop productivity while simultaneously mitigating the environmental consequences of intense management. This study examined the simultaneous response of nitrate nitrogen (NO3-N) leaching losses and maize (Zea mays L.) yield to varied fertilizer N management using field observations and the Integrated BIosphere Simulator (IBIS) model. The model was validated against six years of field observations in chisel-plowed maize plots receiving an optimal (180 kg N ha(-1)) fertilizer N application and in N-unfertilized plots on a silt loam soil near Arlington, Wisconsin. Predicted values of grain yield, harvest index, plant N uptake, residue C to N ratio, leaf area index (LAI), grain N, and drainage were within 20% of observations. However, simulated NO3-N leaching losses, NO3-N concentrations, and net N mineralization exhibited less interannual variability than observations, and had higher levels of error (20-65%). Potential effects of 30% higher (234 kg N ha(-1)) and 30% lower (126 kg N ha(-1)) fertilizer N use (from optimal) on NO3-N leaching loss and maize yield were simulated. A 30% increase in fertilizer N use increased annual NO3-N leaching by 56%, while yield increased by only 1%. The NO3-N concentration in the leachate solution at 1.4 m below the soil surface was 30.7 mg L(-1). When fertilizer N use was reduced by 30% (from optimal), annual NO3-N leaching losses declined by 42% after seven years, and annual average yield only decreased by 8%. However, NO3-N concentration in the leachate solution remained above 10 mg L(-1) (11.3 mg L( 1)). Clearly, nonlinear relationships existed between changes in fertilizer use and NO3-N leaching losses over time. Simulated changes in NO3-N leaching were greater in magnitude than fertilizer N use changes. PMID- 12549566 TI - Methane emissions of beef cattle on forages: efficiency of grazing management systems. AB - Fermentation in the rumen of cattle produces methane (CH4). Methane may play a role in global warming scenarios. The linking of grazing management strategies to more efficient beef production while reducing the CH4 emitted by beef cattle is important. The sulfur hexafluoride (SF6) tracer technique was used to determine the effects of best management practices (BMP) grazing compared with continuous grazing on CH4 production in several Louisiana forages during 1996-1998. Cows and heifers (Bos taurus) grazed common bermudagrass [Cynodon dactylon (L.) Pers.], bahiagrass (Paspalum notatum Flugge), and ryegrass (Lolium multiflorum Lam.) pastures and were wintered on bahiagrass hay with supplements of protein molasses blocks (PMB), cottonseed meal and corn (CSMC), urea and corn (URC), or limited ryegrass grazing (LRG). Daily CH4 emissions were between 89 and 180 g d(-1) for young growing heifers and 165 to 294 g d(-1) for mature Simbrah cows. Heifers on "ad lib" ryegrass in March and April produced only one-tenth the CH4 per kg of gain as heifers on LRG of 1 h. Using BMP significantly reduced the emission of CH4 per unit of animal weight gain. Management-intensive grazing (MIG) is a BMP that offers the potential for more efficient utilization of grazed forage crops via controlled rotational grazing and more efficient conversion of forage into meat and milk. Projected CH4 annual emissions in cows reflect a 22% reduction from BMP when compared with continuous grazing in this study. With the BMP application of MIG, less methane was produced per kilogram of beef gain. PMID- 12549567 TI - Spatial variability of soil carbon in forested and cultivated sites: implications for change detection. AB - The potential to sequester atmospheric carbon in agricultural and forest soils to offset greenhouse gas emissions has generated interest in measuring changes in soil carbon resulting from changes in land management. However, inherent spatial variability of soil carbon limits the precision of measurement of changes in soil carbon and hence, the ability to detect changes. We analyzed variability of soil carbon by intensively sampling sites under different land management as a step toward developing efficient soil sampling designs. Sites were tilled cropland and a mixed deciduous forest in Tennessee, and old-growth and second-growth coniferous forest in western Washington, USA. Six soil cores within each of three microplots were taken as an initial sample and an additional six cores were taken to simulate resampling. Soil C variability was greater in Washington than in Tennessee, and greater in less disturbed than in more disturbed sites. Using this protocol, our data suggest that differences on the order of 2.0 Mg C ha(-1) could be detected by collection and analysis of cores from at least five (tilled) or two (forest) microplots in Tennessee. More spatial variability in the forested sites in Washington increased the minimum detectable difference, but these systems, consisting of low C content sandy soil with irregularly distributed pockets of organic C in buried logs, are likely to rank among the most spatially heterogeneous of systems. Our results clearly indicate that consistent intramicroplot differences at all sites will enable detection of much more modest changes if the same microplots are resampled. PMID- 12549568 TI - Wet chemical and phosphorus-31 nuclear magnetic resonance analysis of phosphorus speciation in a sandy soil receiving long-term fertilizer or animal manure applications. AB - In areas under intensive livestock farming and with high application rates of animal manure, inorganic and organic phosphorus (P) may be leached from soils. Since the contribution of these P compounds to P leaching may differ, it is important to determine the speciation of P in these soils. We determined the effect of various fertilization regimes on the P speciation in NaOH-Na2EDTA (ethylenediaminetetraacetic acid) and water extracts of acidic sandy soil samples from the top 5 cm of grassland with wet chemical analysis and 31P nuclear magnetic resonance (NMR) spectroscopy. These soils had been treated for a period of 11 years with no fertilizer (control), N (no P application), N-P-K, or different animal manures. Inorganic P was highly elevated in the NaOH-Na2EDTA extracts of the soils amended with N-P-K or animal manures, while organic P increased only in the soil treated with pig slurry. Water-extractable P showed a similar trend. As indicated by 31P NMR, orthophosphate monoesters were the main organic P compounds in all soils. Our results suggest that long-term applications of large amounts of P fertilizer and animal manures caused an accumulation of inorganic P, resulting in an increase of the potential risk related to mobilization of inorganic P in the top 5 cm of these soils. PMID- 12549569 TI - Evaluation of attachment of Cryptosporidium parvum and Giardia lamblia to soil particles. AB - Transport of Cryptosporidium parvum oocysts and Giardia lamblia cysts in the aquatic environment is poorly understood. Information about their transport is essential for actual risk assessment and development of effective control practices. Several studies have suggested that attachment to soil particles is not likely to occur, but the hypothesis has not been well tested, partly because enumeration of C. parvum oocysts or G. lamblia cysts [written as (oo)cysts] in the presence of soil has been difficult. In this paper, a combination of flow cytometry, and epifluorescence and confocal microscopy was successfully used to enumerate (oo)cysts in the presence of soil and determine whether (oo)cysts travel freely in water or attached to soil particles. The maximum soil concentration in water samples for reliable enumeration of (oo)cysts was 2 mg/L. Particle attachment experiments detected attached pairs of oppositely charged beads and (oo)cysts, while no attachment was observed between like charged beads, (oo)cysts, and soil particles. These results support the hypothesis that electrostatic forces govern the interaction between (oo)cysts and soil particles. Batch experiments further confirmed the null hypothesis (p > 0.05) that (oo)cysts do not attach to natural soil particles. These findings suggest that, when (oo)cysts have been entrained in overland flow (i.e., runoff), they will travel freely in the water and not as part of the particulate sediment load. PMID- 12549570 TI - Putative temporal variability of Escherichia coli ribotypes from yearling steers. AB - Escherichia coli is a ubiquitous component of the intestinal microflora of warm blooded animals, and is an indicator of fecal contamination of surface waters. Ribotype profiling of E. coli is one of several genotypic methods that has been developed to determine the host origin of fecal bacteria. Like most genotypic methods of source tracking, ribotyping requires a host origin database to identify environmental isolates. To determine the extent of temporal variability of ribotypes and its effect on a host origin database, E. coli isolates were obtained from fecal samples of two herds of Black Angus steers at a long-term experimental site at four sampling times from October 1999 to July 2000. Fecal samples were taken from six randomly chosen steers at each time. At a similarity index of 90% as calculated by unweighted pair-group method using arithmetic averages (UPGMA), 240 ribotypes were identified from 451 E. coli isolates. Only 20 ribotypes (8.3%), comprising 33% of the total isolates, were shared among sampling times and were considered resident ribotypes. Two of the twenty resident ribotypes appeared at three sampling times, and the remaining eighteen appeared at two. The majority of the ribotypes, therefore, were transient and unique to each sampling time and steer. Both the apparent turnover of E. coli ribotypes and a clonal diversity index of 0.97 (indicative of extensive ribotype variability) suggest the necessity of ribotyping a large number E. coli isolates per host to establish a host origin database that is independent of temporal variability, or complete enough to be effective. PMID- 12549571 TI - Relationships between phosphorus levels in soil and in runoff from corn production systems. AB - Phosphorus-enriched runoff from cropland can hasten eutrophication of surface waters. A soil P level exceeding crop needs due to long-term fertilizer and/or manure applications is one of several potential sources of increased P losses in runoff from agricultural systems. Field experiments were conducted at locations representative of three major soil regions in Wisconsin in corn (Zea mays L.) production systems to determine the effect of tillage, recent manure additions, soil P extraction method, and soil sampling depth (0-2, 0-5, and 0-15 cm) on the relationship between soil test P level and P concentrations in runoff. Runoff from simulated rainfall (75 mm h(-1)) was collected from 0.83-m2 areas for 1 h after rainfall initiation and analyzed for dissolved phosphorus (DP), total phosphorus (TP), and sediment. The DP fraction of the TP concentration in runoff ranged from 5 to 17% among sites with most of the variation in TP due to varying sediment concentration on the well-drained silt loam soils and to soil test P level on the poorly drained silty clay loam soil. In 213 observations across a range of soils and managements, good relationships occurred between soil test P level and DP concentration in runoff for most of the tests and sampling depths used. Recent manure additions and high levels of surface cover from corn residue sometimes masked this relationship. The slope of DP relative to soil test P level was markedly higher on the silty clay loam soil than on the silt loam soils possibly due to soil permeability-infiltration rate differences. Agronomic soil P tests were as effective as environmentally oriented soil P tests for predicting DP concentrations in runoff. PMID- 12549572 TI - Microbial reduction of hexavalent chromium under vadose zone conditions. AB - Hexavalent chromium [Cr(VI)] is a common contaminant associated with nuclear reactors and fuel processing. Improper disposal at facilities in and and semiarid regions has contaminated underlying vadose zones and aquifers. The objectives of this study were to assess the potential for immobilizing Cr(VI) using a native microbial community to reduce soluble Cr(VI) to insoluble Cr(III) under conditions similar to those in the vadose zone, and to evaluate the potential for enhancing biological Cr(VI) reduction through nutrient addition. Batch microcosm and unsaturated flow column experiments were performed. Native microbial communities in subsurface sediments with no prior Cr(VI) exposure were shown to be capable of Cr(VI) reduction. In both the batch and column experiments, Cr(VI) reduction and loss from the aqueous phase were enhanced by adding high levels of both nitrate (NO3-) and organic C (molasses). Nutrient amendments resulted in up to 87% reduction of the initial 67 mg L(-1) Cr(VI) in an unsaturated batch experiment. Molasses and nitrate additions to 15 cm long unsaturated flow columns receiving 65 mg L(-1) Cr(VI) resulted in microbially mediated reduction and immobilization of 10% of the Cr during a 45-d experiment. All of the immobilized Cr was in the form of Cr(III), as shown by XANES analysis. This suggests that biostimulation of microbial Cr(VI) reduction in vadose zones by nutrient amendment is a promising strategy, and that immobilization of close to 100% of Cr contamination could be achieved in a thick vadose zone with longer flow paths and longer contact times than in this experiment. PMID- 12549573 TI - Patterns of short-term sedimentation in a freshwater created marsh. AB - This study investigated different sedimentation measurement techniques and examined patterns of short-term sedimentation in two 1-ha replicate created freshwater marshes in central Ohio, USA. Short-term (one-year) sediment accumulation above feldspar, clay, glitter, and sand artificial marker horizons was compared at different water depths and distances from wetland inflow. A sediment budget was also constructed from turbidity and suspended sediment data for comparison with marker horizons. Glitter and sand marker horizons were the most successful for measuring sediment accumulation (81-100% marker recovery), while clay markers were completely unsuccessful. The sedimentation rate for both wetlands averaged 4.9 cm yr(-1) (36 kg m(-2) yr(-1)), and ranged from 1.82 to 9.23 cm yr(-1) (12.4 to 69.7 kg m(-2) yr(-1)). Sedimentation rates in deep, open water areas were significantly higher than in shallow, vegetated areas for both wetlands (t test, p < 0.05). However, observed sedimentation patterns may be attributed more to preferential flow through open water areas than to water depth or presence of vegetation. Contrary to the expected spatial distribution, sedimentation was highly variable within the wetlands, suggesting that bioturbation and turbulence may cause significant resuspension or that high hydrologic loads may distribute sediments throughout the basins. A sediment budget estimated sediment retention of approximately 740 g m(-2) yr(-1) per wetland (43% removal rate), yet gross sediment accumulation was 36,000 g m(-2) yr(-1) measured by marker horizons. These results suggest that erosive forces may have influenced sedimentation, but also may indicate problems with the sediment budget calculation methodology. PMID- 12549574 TI - Spatial analysis of phosphorus sorption capacity in a semiarid altered wetland. AB - The observed increase in phosphorus (P) loading into the Jordan River could increase eutrophication processes in Lake Kinneret, the only freshwater lake in Israel, which provides 25% of the country's drinking water. The P may originate from the peat soils of the highly altered Hula Valley's semiarid wetland ecosystem through which the Jordan River runs. The objectives of this research were to ascertain the sorption capacity of these soils and to identify areas with high potential for P release from soils to ground water. We extracted 80 soil samples collected across the valley with ammonium oxalate and determined the ratio of extractable P to Fe and Al, from which we derived the degree of phosphorus saturation (DPS). A relatively low DPS (<15%) was observed in Histosols compared with the high DPS (>30%) observed in many of the hydromorphic organo-mineral soils. We used a sequential Gaussian simulation technique to assess the spatial pattern of the DPS and found that the Histosols have a low probability (<10%) of exceeding the widely used environmental DPS threshold of 25%. The areas characterized by mineral soils, such as hydromorphic Vertisols and various marl redoximorphic soils, have a high probability (>60%) of exceeding the threshold value. The ability to predict the concentrations of dissolved P in ground water based on DPS values was somewhat impaired because of the preferential flow characteristics in this altered wetland. PMID- 12549575 TI - Decadal change in vegetation and soil phosphorus pattern across the Everglades landscape. AB - Wetlands respond to nutrient enrichment with characteristic increases in soil nutrients and shifts in plant community composition. These responses to eutrophication tend to be more rapid and longer lasting in oligotrophic systems. In this study, we documented changes associated with water quality from 1989 to 1999 in oligotrophic Everglades wetlands. We accomplished this by resampling soils and macrophytes along four transects in 1999 that were originally sampled in 1989. In addition to documenting soil phosphorus (P) levels and decadal changes in plant species composition at the same sites, we report macrophyte tissue nutrient and biomass data from 1999 for future temporal comparisons. Water quality improved throughout much of the Everglades in the 1990s. In spite of this improvement, though, we found that water quality impacts worsened during this time in areas of the northern Everglades (western Loxahatchee National Wildlife Refuge [NWR] and Water Conservation Area [WCA] 2A). Zones of high soil P (exceeding 700 mg P kg(-1) dry wt. soil) increased to more than 1 km from the western margin canal into the Loxahatchee NWR and more than 4 km from northern boundary canal into WCA-2A. This doubling of the high soil P zones since 1989 was paralleled with an expansion of cattail (Typha spp.)-dominated marsh in both regions. Macrophyte species richness declined in both areas from 1989 to 1999 (27% in the Loxahatchee NWR and 33% in WCA-2A). In contrast, areas well south of the Everglades Agricultural Area, induding WCA-3A and Everglades National Park (ENP), did not decline during this time. We found no significant decadal change in plant community patterns from 1989 and 1999 along transects in southern WCA-3A or Shark River Slough (ENP). Our 1999 sampling also included a new transect in Taylor Slough (ENP), which will allow change analysis here in the future. Regular sampling of these transects, to verify decadal-scale environmental impacts or improvements, will continue to be an important tool for long-term management and restoration of the Everglades. PMID- 12549576 TI - Visualizing bromide and iodide water tracer in soil profiles by spray methods. AB - In this study we developed and tested a spray method to visualize bromide water tracer in soil profiles. The method is based on the transformation reaction of a white precipitate into a colored one (Prussian blue) in the presence of Br-. After application of water containing bromide (0.2-0.4% wt.), a soil profile is dug out from the irrigated area and sprayed with a Br- indication suspension containing ferric ion and silver ferrocyanide precipitate. About two hours later, the pattern of irrigation water movement in the soil profile appears due to the formation of Prussian blue complex. We describe the method and demonstrate its use in a field experiment to visualize water flow paths. Since this method might be subject to possible interference from Cl-, a newly designed method with iodide ion as a water tracer and its indication solution containing soluble starch and ferric ion is also presented and recommended for use in soils with high chloride background. PMID- 12549577 TI - Enumeration of waterborne Escherichia coli with petrifilm plates: comparison to standard methods. AB - Escherichia coli is often monitored in environmental waters as an indicator of the possible presence of human pathogens associated with feces. Petrifilm E. coli/coliform count plates (3M, Minneapolis, MN), previously validated for enumerating E. coli in food, were tested for monitoring E. coli in environmental water. Escherichia coli counts in environmental water samples enumerated with Petrifilm were significantly correlated (R > 0.9; slope = 0.9-1.0; p < 0.001) with counts obtained with three commonly used methods, mTEC (Becton Dickinson, Sparks, MD), m-ColiBlue (Hach, Loveland, CO), and Colilert-18/IDEXX Quanti-Tray 2000 (IDEXX, Westbrook, ME). Blue colonies on Petrifilm plates were most reliably identified as E. coli when accompanied by gas formation, as determined by characterization of the colonies on MacConkey agar plates (PML Microbiologicals, Mississauga, ON, Canada) and by polymerase chair reaction (PCR) with E. coli specific primers. The main disadvantage of Petrifilm plates for environmental water testing is the small volume (1 mL per sample) that can be tested; however, the plates appear to be suitable for screening and locating sites that exceed criteria for total body and partial body contact. Simplicity of use and storage, reliability, and relatively low cost make Petrifilm plates suitable for volunteer based and educational water quality monitoring applications, particularly when used as a preliminary screening method to identify problem sites. PMID- 12549578 TI - Inferred goals in narratives: evidence from self-paced reading, recall, and eye movements. AB - Three convergent methodologies were used to investigate the generation and reinstatement of goals not explicitly stated in a text. Readers read paragraphs adapted from J. S. Huitema, S. Dopkins, C. M. Klin, & J. L. Myers's (1993) study, which conveyed a character's goal early in the text. The goal was either stated explicitly or implied. An event was described later in the text that was either consistent or inconsistent with the goal. Line-by-line reading data, recall for the narratives, and eye-movement data were collected. Evidence is presented that readers infer a character's goal online at the time the information is presented, and the inferred goal functions like an explicitly stated goal in online comprehension processes and in the resulting memory representation. PMID- 12549579 TI - Auditory registration without learning. AB - In 3 studies, the authors explore how repeated exposure to a spoken word affects memory for perceptual attributes associated with the word (such as a talker's voice or a word's plurality). Subjects heard a list of words; particular words were repeated differing numbers of times. At test, subjects estimated the frequency of each word, with instructions to give frequency judgments of "zero" to words with changed attributes. The experiments demonstrate that memory for perceptual attributes improves very little after the first few repetitions, although word memory continues to improve. The experiments extend the registration without learning effect (D. L. Hintzman, T. Curran, & B. Oppy, 1992) to auditory words, to complex attributes (voice), and to conditions of low and high stimulus variability (two or many voices). PMID- 12549580 TI - Encoding fluency is a cue used for judgments about learning. AB - The authors used paired-associate learning to investigate the hypothesis that the speed of generating an interactive image (encoding fluency) influenced 2 metacognitive judgments: judgments of learning (JOLs) and quality of encoding ratings (QUEs). Results from Experiments 1 and 2 indicated that latency of a keypress indicating successful image formation was negatively related to both JOLs and QUEs even though latency was unrelated to recall. Experiment 3 demonstrated that when concrete and abstract items were mixed in a single list, latency was related to concreteness, judgments, and recall. However, item concreteness and fluency influenced judgments independently of one another. These outcomes suggest an important role of encoding fluency in the formation of metacognitive judgments about learning and future recall. PMID- 12549581 TI - Effective warnings in the Deese-Roediger-McDermott false-memory paradigm: the role of identifiability. AB - These experiments document that warnings can substantially reduce false memories in the Deese-Roediger-McDermott (DRM) paradigm when the critical items are easily identifiable. Participants in a norming study identified the critical item after hearing a list of words. The lists with critical items that could be identified by the largest proportion of participants (high identifiable [HI] lists) and the smallest proportion of participants (low identifiable [LI] lists) were used in the experiment. Participants heard lists of words (e.g., bed, rest, doze) related to a critical item (e.g., sleep) and were warned about the nature of the lists before the study phase. The results indicated that warnings reduced false recognition of critical items for HI lists but not LI lists. PMID- 12549582 TI - Spreading activation or spooky action at a distance? AB - How do preexisting connections among a word's associates facilitate its cued recall and recognition? A spreading-activation model assumes activation spreads to, among, and from a studied word's associates, and that its return is what strengthens its representation. An activation-at-a-distance model assumes strengthening is produced by the synchronous activation of the word's associates. The spread model predicts that connections among the studied word's associates will have a greater effect on memory when more of its associates return activation. The distance model predicts that total connections are important, not their direction. The results of cued recall experiments supported the distance model in showing that that connections among the associates facilitated recall regardless of the number of returning connections. PMID- 12549583 TI - Take the best or look at the rest? Factors influencing "one-reason" decision making. AB - Aspects of an experimental environment were manipulated in 3 experiments to examine the parameters under which the "take-the-best" (TTB) heuristic (e.g., G. Gigerenzer & D. G. Goldstein, 1996) operates. Results indicated TTB use to be more prevalent when the cost of information was high, when validities of the cues were known, and when a deterministic environment was used. However, large individual variability in strategy use was observed as well as a significant proportion of behavior inconsistent with TTB, expecially its stopping rule. The results demarcate some of the heuristic's boundary conditions and also question the validity of TTB as a psychologically plausible and pervasive model of behavior. PMID- 12549584 TI - Concave utility, transaction costs, and risk in measuring discounting of delayed rewards. AB - Research has consistently found that the decline in the present values of delayed rewards as delay increases is better fit by hyperbolic than by exponential delay discounting functions. However, concave utility, transaction costs, and risk each could produce hyperbolic-looking data, even when the underlying discounting function is exponential. In Experiments 1 (N = 45) and 2 (N = 103), participants placed bids indicating their present values of real future monetary rewards in computer-based 2nd-price auctions. Both experiments suggest that utility is not sufficiently concave to account for the superior fit of hyperbolic functions. Experiment 2 provided no evidence that the effects of transaction costs and risk are large enough to account for the superior fit of hyperbolic functions. PMID- 12549585 TI - An associative framework for probability judgment: an application to biases. AB - Three experiments show that understanding of biases in probability judgment can be improved by extending the application of the associative-learning framework. In Experiment 1, the authors used M. A. Gluck and G. H. Bower's (1988a) diagnostic-learning task to replicate apparent base-rate neglect and to induce the conjunction fallacy in a later judgment phase as a by-product of the conversion bias. In Experiment 2, the authors found stronger evidence of the conversion bias with the same learning task. In Experiment 3, the authors changed the diagnostic-learning task to induce some conjunction fallacies that were not based on the conversion bias. The authors show that the conjunction fallacies obtained in Experiment 3 can be explained by adding an averaging component to M. A. Gluck and G. H. Bower's model. PMID- 12549586 TI - Causal beliefs and conditioned responses: retrospective revaluation induced by experience and by instruction. AB - The author tested causal beliefs and conditioned responses in a task involving retrospective revaluation of the causal status of a target cue with respect to electric shock. Successful revaluation was observed on both self-report shock expectancy and skin conductance, whether the training trials were directly experienced, described, or partly experienced and partly described. The results contradict models that link anticipatory conditioned responses to a separate or earlier process from that underlying explicit causal knowledge. They suggest instead that a single learning process gives rise to propositional knowledge that (a) drives anticipatory responding, (b) forms the basis for self-reported causal beliefs, and (c) can be combined with other knowledge, provided either by experience or symbolically, to generate inferences such as retrospective revaluation. PMID- 12549587 TI - Classification of exemplars with single- and multiple-feature manifestations: the effects of relevant dimension variation and category structure. AB - Most classification research focuses on cases in which each abstract feature has the same surface manifestation whenever it is presented. Previous research finds that people have difficulty learning to classify when each abstract feature has multiple surface manifestations. These studies created multiple manifestations by varying aspects of the stimuli irrelevant to the abstract feature dimension. In this article, multiple manifestations were created by varying aspects of the stimuli relevant to the abstract feature dimension. People given categories with the family resemblance category structure often used in psychology experiments had difficulty learning to classify when multiple manifestations were present, even though the variation was relevant. This effect was reversed when a family resemblance structure with nondiagnostic values was used. PMID- 12549588 TI - Thematic role focusing by participle inflections: evidence from conceptual combination. AB - The authors examined how people integrate knowledge of agents and patients of events with the temporal and causal properties of present and past participles to constrain interpretation of isolated participle-noun phrases like arresting cop and arrested crook. Good-agent head nouns were more easily combined with present participles (e.g., arresting cop) than with past participles (e.g., arrested cop), and the reverse was true for good patients. Furthermore, present-participle good-patient phrases (e.g., serving customer) were often interpreted as verb phrases. This research provides further evidence of the interaction between morphosyntactic cues and world knowledge of events in language comprehension. PMID- 12549589 TI - The influence of morphological information on cataphoric pronoun assignment. AB - In 3 eye-tracking experiments, the authors investigated the use of morphological information during pronoun resolution. Experiments 1 and 2 showed that disruption occurred when a preferred assignment was inconsistent with gender information. Experiment 2 ensured that this difficulty was not due to the introduction of a new discourse entity. Experiment 3 showed that disruption also occurred when number information was inconsistent with the preferred assignment. The results indicate that the use of morphological information is delayed until after the computation of coreference relations. PMID- 12549590 TI - A critique of Oaksford, Chater, and Larkin's (2000) conditional probability model of conditional reasoning. AB - M. Oaksford, N. Chater, and J. Larkin (2000) proffered a Bayesian model in which conditional inferences are a direct function of conditional probabilities. In the current article, the authors first considered this model regarding the processing of negatives in conditional reasoning. Its predictions were evaluated against a large-scale meta-analysis (W. J. Schroyens, W. Schaeken, & G. d'Ydewalle, 2001b). This evaluation shows that the model is flawed: The relative size of the negative effects does not match predictions. Next, the authors evaluated the model in relation to inferences about affirmative conditionals, again considering the results of a meta-analysis (W. J. Schroyens, W. Schaeken, & G. d'Ydewalle, 2001a). The conditional probability model is countered by the data reported in literature; a mental models based model produces a better fit. The authors conclude that a purely probabilistic model is deficient and incomplete and cannot do without algorithmic processing assumptions if it is to advance toward a descriptively adequate psychological theory. PMID- 12549591 TI - Preoperative albumin and surgical site identify surgical risk for major postoperative complications. AB - BACKGROUND: Although malnutrition contributes to morbidity, studies of pre- and postoperative nutrition often include well-nourished patients unlikely to benefit from therapy and usually do not stratify by the site of surgical pathology. This study evaluates whether perceived preoperative markers of nutritional status recorded in charts correlates with postoperative complications and resource use in patients who receive no preoperative nutrition support and reinterprets the results of several conflicting randomized, prospective studies in this context. METHODS: This is a retrospective cohort study of 526 surgical patients who had preoperative serum albumin levels measured and were undergoing elective esophageal, gastric, pancreaticoduodenal, or colon surgery between 1992 and 1996 who could have received preoperative nutrition but did not. RESULTS: Most medical records contained inadequate analysis of preoperative nutritional status, but preoperative albumin correlated inversely with complications, length of stay, postoperative stay, intensive care unit (ICU) stay, mortality, and resumption of oral intake. Patients undergoing esophageal or pancreatic procedures sustained a significantly higher complication rate at most albumin levels, whereas colonic surgery resulted in lower complication rates at the same albumin levels. Resource use (eg, length of stay and ICU stay) related to these complication rates; esophageal and pancreatic procedures used the most resources and colon procedures used the fewest at most albumin levels. This lack of appreciation for nutritional risk and operative site can explain discrepancies in outcome noted in several randomized, prospective nutritional studies and must be applied to the design and implementation of new studies. CONCLUSIONS: Elective, non-emergent esophageal and pancreatic procedures performed in patients who could have had surgery delayed for preoperative nutrition, but did not, result in higher risk than colon surgery at any given level of serum albumin below 3.25 g/dL. Patient populations in trials should be stratified by operative site and by markers of nutritional status. Degree of hypoalbuminemia and other potential markers of nutritional status may explain many of the discrepancies between trials of nutrition support. Preexisting hypoalbuminemia in patients undergoing elective surgery remains underappreciated, unrecognized, and untreated in many hospitalized patients. PMID- 12549592 TI - Is serum albumin an independent predictor of survival in patients with breast cancer? AB - BACKGROUND: The medical literature shows several examples of an inverse relationship between serum albumin levels and survival in patients with advanced cancer. METHODS: We investigated the effect of baseline serum levels on 180 consecutively treated patients who were diagnosed with breast cancer from March 1993 to December 1999 at our institution. We investigated the effect of low levels of serum albumin (<3.5 g/dL) using univariate and the multivariate Cox regression model. RESULTS: Demographically, the median age of these patients was 51 years of age, with a range of 28 to 88 years. In this series, 28 patients have died and 9 have developed a recurrence of tumor. The patient distribution by tumor stage was 53, stage 1; 47, stage 2A; 29, stage 2B; 11, stage 3; 13, stage 4A; and 21, stage 4B. In this cohort, the overall 5-year survival was 69%. The Cox regression analysis found that normal levels of albumin (>3.5 g/dL) reduced the risk of death by 72% (p = .0033). Only tumor stage had a larger impact on survival (p < .0000). Other factors achieving statistical significance were overexpression of the HER2/Neu antigen (p = .0167) and breast antigen (p = .0116). CONCLUSION: Univariate statistical analysis found that low levels of serum albumin adversely affected survival by a statistically significant level for all stages of breast cancer. We found that a baseline serum albumin level was a powerful prognostic variable, which accounted for 7.35% of the variation in patient survival time. PMID- 12549593 TI - Achievement of steady state optimizes results when performing indirect calorimetry. AB - BACKGROUND: The use of steady state as the endpoint for performance of indirect calorimetry (IC) is controversial. We designed this prospective study to evaluate the necessity and significance of achieving steady state. METHODS: Patients with respiratory failure placed on mechanical ventilation in a short- or long-term acute care unit at any 1 of 3 university-based urban hospitals were eligible for the study. The 24-hour total energy expenditure (TEE) was determined by a Nellcor Puritan Bennett 7250 continuous IC monitor. Measured gas exchange parameters were obtained and averaged every 1 minute for the initial hour and then every 15 minutes for the next 23 hours. Over the initial hour, resting energy expenditure (REE) was averaged for intervals over the first 20, 30, 40, and 60 minutes, and for various definitions of steady state where oxygen consumption (VO2) and carbon dioxide production (VCO2) changed by <10%, 15%, and 20%. Coefficient of variation (CV) was calculated for VO2 over the first 30 minutes of study. RESULTS: Twenty two patients (mean age, 52.8 years, 59% male, mean Acute Physiology and Chronic Health Evaluation (APACHE III) score 42.0) were entered in the study. The best correlation between short-term "snapshot" REE and the 24-hour TEE was achieved by the steady-state period defined by the most stringent criteria (change in VO2 and VCO2 by <10%). The average REE for all steady-state and interval periods correlated significantly to TEE with no significant difference in the absolute values for REE and TEE. Adding 10% for an activity factor to the average REE for each steady-state and interval period again correlated to TEE in a similar fashion with the same R value, but the absolute values for REE + 10% for all steady-state and interval periods were significantly different than the corresponding TEE. In those patients with less variation (CV for VO2 < or = 9.0), the REE obtained for the steady-state period defined by the most stringent criteria still had the best correlation, but similar correlation could be obtained by interval testing of > or = 30-minute duration. In those patients with greater variation (CV for VO2 >9.0), interval testing of at least 60 minutes or more was required to attain levels of correlation similar to that achieved by the steady-state period defined by the most stringent criteria. CONCLUSIONS: These data support the use of steady state, best defined as an interval of 5 consecutive minutes whereby VO2 and VCO2 change by <10%. The mean REE from this period correlates best to the 24-hour TEE regardless of CV. IC testing can be completed after achievement of steady state. Activity factors of 10% to 15% should not be added to the steady-state REE, because this practice significantly decreases the accuracy. In patients who fail to achieve steady state, the CV helps to determine the appropriate duration of IC testing. In those patients with a low CV (< or = 9.0), 30-minute test duration is adequate. In patients with CV >9.0, test duration of at least 60 minutes may be required. These latter patients should be considered for 24-hour IC testing. PMID- 12549594 TI - Clinical use of the respiratory quotient obtained from indirect calorimetry. AB - BACKGROUND: The respiratory quotient (RQ) obtained from indirect calorimetry (IC), defined by the ratio carbon dioxide production (VCO2)/oxygen consumption (VO2), is affected by extremes of substrate use by the body. Underfeeding, which promotes use of endogenous fat stores, should cause decreases in the RQ, whereas overfeeding, which results in lipogenesis, should cause increases in the RQ. Marked increases in VCO2 (with subsequent increases in RQ) in response to overfeeding may cause respiratory compromise in patients with limited pulmonary reserve. Thus, variation in the RQ in response to the feeding regimen may indicate inappropriate feeding and serve as a marker for patient intolerance. This prospective, multicenter study was designed to determine the clinical use of RQ for monitoring adequacy and tolerance of nutrition support. METHODS: Patients in any 1 of 30 long-term acute care Kindred hospitals made nil orally (NPO) and placed on total parenteral or enteral feeding were eligible for this study. Arterial blood gas, serum ketones, 24-hour collection of urine urea nitrogen, and IC measurements were obtained on all. Actual volume of enteral/parenteral feeding infused over the 24 hours before performance of IC was documented. RESULTS: A total of 263 patients (mean age, 70.2 years, 57.4% male) were entered in the study. Of the 263 study patients, 88.6% required mechanical ventilation, and 92.0% received enteral tube feeding only. Overall, 41.5% of patients were overfed, receiving >110% of required calories, whereas 34.2% were underfed, receiving <90% of required calories. The ratio of calories provided/required correlated significantly with overall measured RQ (p < .0001; R2 = .16). Correcting for the metabolism of protein by calculating a nonprotein RQ (NPRQ) from a 24-hour urine urea nitrogen did not improve this correlation (p < .0001, R2 = .32). Using a measured NPRQ >1.0 to identify overfeeding had an acceptable specificity of 85.1% but a low sensitivity of 38.5%. Similarly, use of a NPRQ <0.85 to determine underfeeding had a specificity of 72.2% and a sensitivity of 55.8%. Comparing the measured NPRQ with a predicted reference RQ (based on percent infusion of carbohydrate/fat and the Lusk table) did not improve the overall use of RQ. In the majority of patients (67.7%), comparison of the measured NPRQ to the predicted value failed to differentiate appropriate (meeting 100% +/- 10% of requirements) from inappropriate feeding. Deviation of the measured NPRQ from predicted failed to identify factors unrelated to substrate use purported to affect the RQ (such as acid/base disturbances or hyper/hypoventilation). Increasing measured RQ did correlate significantly with increasing respiratory rate (p = .002, R2 = .04) and decreasing tidal volume (p = .002, R2 = .04), suggesting reduced tolerance with development of shallow rapid respirations and ventilatory compromise. CONCLUSIONS: Although changes in the overall and nonprotein RQ correlate to percent calories provided/required, low sensitivity and specificity limit its efficacy as an indicator of over- or underfeeding. The RQ should not be used to finely adjust the nutrition support regimen. Elevation of overall measured RQ > or = 1.0 may be associated with reduced tolerance and mild respiratory compromise. The clinical use of RQ is limited to a marker of test validity (to confirm measured RQ values are in physiologic range) and a marker for respiratory tolerance of the nutrition support regimen. PMID- 12549595 TI - Effect of neuromuscular blockade on energy expenditure in patients with severe head injury. AB - BACKGROUND: The purpose of this study was to determine the effect of neuromuscular blockade on energy expenditure in severely head-injured patients; to determine the effects of body temperature, nutrition support, and morphine use on metabolic rate; and to compare measured energy expenditure with values from predictive equations. METHODS: Energy expenditure was measured using indirect calorimetry in 2 groups of ventilated patients-18 with severe head injury during and after administration of pancuronium bromide and morphine, and second, 14 severely traumatized patients without severe head injury (trauma group) who received morphine without neuromuscular blockade. RESULTS: The mean energy expenditure of head-injured patients increased significantly once pancuronium was discontinued, ie, from 24.2 +/- 3.1 to 28.7 +/- 4.6 kcal/kg (p = .002). This effect was independent of other relevant variables such as morphine dose, body temperature, and nutrition support. When compared with the Harris-Benedict and World Health Organization predictive equations, neuromuscular blockade resulted in a stress factor of only 0.96 and 0.95, respectively, which increased to 1.19 and 1.18, respectively, once blockade was discontinued. Head-injured patients not on neuromuscular blockade had a significantly greater energy expenditure when compared with the trauma group (p = .02). CONCLUSIONS: Neuromuscular blockade in severely head-injured patients decreases energy expenditure to basal levels, independent of morphine use, body temperature, and feeding. Levels of hypermetabolism in both the head-injured and trauma groups were relatively low, at 19% and 5% above predicted values, respectively. This study provides useful information for the management of nutrition support in severely traumatized patients. PMID- 12549596 TI - Increased resting energy expenditure in patients with end-stage renal disease. AB - BACKGROUND: Protein-calorie malnutrition is a significant problem for patients with end-stage renal disease. Increased resting energy expenditure may be an important contributing factor. We postulate that resting energy expen diture in the different stages of renal disease and treatments may be different. METHODS: Resting energy expenditure was measured using a whole-room indirect calorimeter (metabolic chamber) along with nutritional parameters and body composition after 12-hour fasting in 15 patients with advanced chronic renal failure patients, 15 patients on chronic hemodialysis, and 10 patients on peritoneal dialysis. Patients on hemodialysis were assessed on a non-dialysis day. A 2-day dietary recall was used to assess energy intake. RESULTS: Resting energy expenditure, adjusted for fat-free mass, was similar in patients on hemodialysis and peritoneal dialysis but significantly higher than in patients with chronic renal failure (p < .05). Resting energy expenditure in all patients were generally higher (10% to 20%) than predicted values using standard equations derived in normal and obese populations, whereas daily energy intake was less (26% to 34%) than energy expenditure for all groups, adjusted for light daily activity. CONCLUSIONS: End-stage renal disease patients displayed increases in resting energy expenditure over the predicted values derived using normal populations. Resting energy expenditure was significantly higher in patients receiving dialysis, regardless of the modality, than patients with chronic renal failure. Daily energy intake was substantially less than required in all patient groups studied, suggesting that patients with renal failure could develop protein calorie malnutrition because of increased resting energy expenditure, which is exacerbated by dialysis. PMID- 12549597 TI - Use of bioelectrical impedance analysis to predict energy expenditure of hospitalized patients receiving nutrition support. AB - BACKGROUND: The energy requirement of a patient receiving nutrition support is typically estimated by calculating the basal energy expenditure (BEE) using the Harris-Benedict equations and multiplying by stress and activity factors. Because fat-free mass (FFM) and fat mass (FM) are important determinants of BEE, we hypothesized that body composition estimates derived from bioelectrical impedance analysis (BIA) could be used to develop predictive equations for resting energy expenditure (REE) that were more accurate than those calculated using the Harris Benedict equations. METHODS: Seventy-six adults referred to the nutrition support service were studied. REE was measured by indirect calorimetry, and single frequency BIA was used to estimate FFM and FM. Using the first 20 male and 20 female patients, predictive equations for REE were developed by multiple regression analysis, using BIA-derived body composition values, age, and gender. The next 36 patients were used to compare the accuracy of these equations with the Harris-Benedict equations in estimating REE. RESULTS: Using BLA-derived body composition values, gender, and age, predictive equations were developed for REE that explained approximately 65% of the variance. Inclusion of other BIA or anthropometric parameters did not improve the equations. When compared with the Harris-Benedict equations, the equations developed in this study were significantly more accurate, providing an REE estimate that was closer to the measured value in about 75% of patients. CONCLUSIONS: These results indicate that BLA-derived body composition estimates may be used to more accurately predict the energy requirements of patients receiving nutrition support than calculations based on the Harris-Benedict equations. PMID- 12549598 TI - Resting energy expenditure in children with cyanotic and noncyanotic congenital heart disease before and after open heart surgery. AB - BACKGROUND: Failure to thrive is a common problem in children with congenital heart disease (CHD). Resting energy expenditure (REE) in cyanotic and noncyanotic children with CHD before and after open heart surgery has hardly been investigated. METHODS: Twenty-nine children younger than 3 years of age with CHD (14 cyanotic and 15 noncyanotic CHD) who were referred for open heart surgery were enrolled. Data on dietary intake, anthropometric measurements, and indirect calorimetry parameters were measured 24 hours before the surgery, (day -1), and on day 5 after surgery. The measured REE was compared with the Schofield and World Health Organization (WHO) REE prediction equations. RESULTS: The mean +/- SD measured REE was similar in the cyanotic and noncyanotic children before and after surgery (before surgery: 57 +/- 13 and 58 +/- 9 kcal/kg per day, respectively; 5 days after surgery: 59 +/- 10 and 62 +/- 10 kcal/kg per day, respectively). Oxygen consumption (VO2) and carbon dioxide production (VCO2) did not change significantly before and after surgery and were similar in both groups. The measured REE for all children on day -1 and day 5 was similar to the calculated REE using the Schofield equation but was significantly different from the calculated REE using the WHO equation (p < .01). CONCLUSIONS: Significant changes in REE, VCO2, and VO2 were not observed before and 5 days after open heart surgery in children with CHD. These parameters (REE, VCO2, and VO2) were also similar in children with cyanotic versus noncyanotic CHD. The Schofield equation is more accurate than the WHO equation in predicting energy needs of children with CHD, but measurement of REE is preferred over calculation of REE. PMID- 12549599 TI - Effect of an 8-month treatment with omega-3 fatty acids (eicosapentaenoic and docosahexaenoic) in patients with cystic fibrosis. AB - BACKGROUND: Supplementation of the diet with eicosapentaenoic acid and docosahexaenoic acid, the main long-chain omega-3 fatty acids in cell membranes, may have beneficial effects in patients with cystic fibrosis. METHODS: A prospective study involving 30 patients and 20 control subjects was carried out; eicosapentaenoic plus docosahexaenoic acid was equal to 1.3% of caloric intake in the cystic fibrosis patients. Our present study included the evaluation of eicosapentaenoic and docosahexaenoic acid incorporation into erythrocyte membranes and biological and clinical effects in response to long-term (8 months) supplementation with fish oil as a source of eicosapentaenoic and docosahexaenoic acids in patients with cystic fibrosis. RESULTS: Baseline erythrocyte membrane fatty acids showed low levels of linoleic acid and eicosapentaenoic acid and mild elevation of 18:3n6, but similar docosahexanoic acid and other fatty acids in cystic fibrosis patients compared with controls. Fish oil supplementation led to a 1.7-fold (p < .05) and 1.3-fold (not significant) increase of eicosapentaenoic acid in erythrocyte membrane phospholipids after 4 and 8 months of supplementation, respectively, and to a 1.67-fold (p < .05) and 1.38-fold (p < .05) increase of docosahexanoic acid, respectively. Along with these changes, there was a progressive decrease of arachidonic acid (from 8.51 to 6.67 g/100 fatty acids at 4 months and 4.83 g/100 fatty acids at 8 months; p < .05) and an increase of linoleic acid (p < .05) in membrane phospholipids. Analysis of inflammatory markers showed a significant decrease of serum immunoglobulin G (IgG) and of alpha-1 antitrypsin (p < .05) concentrations. Pulmonary function testing showed mild but significant improvement of forced expiratory volume (FEV) 1 from 61% +/- 19% to 57% +/- 19% of predicted values (p < .05). The number of days of antibiotic therapy during the study period was markedly lower compared with the preceding 8-month period (392 versus 721 days; p < .05). CONCLUSION: Long-term eicosapentaenoic plus docosahexanoic acid supplementation (8 months) has positive effects, such as decreasing inflammation, in cystic fibrosis. PMID- 12549600 TI - Effects of particle size on blood clearance and tissue uptake of lipid emulsions with different triglyceride compositions. AB - BACKGROUND: Particle size of IV lipid emulsions affects the catabolism of long chain triglyceride (LCT) emulsions, but little is known about its effect on the catabolism of medium-chain triglyceride (MCT)- and fish oil (FO)-containing emulsions. METHODS: Large (VLDL size), intermediate, and small (IDL size) emulsions with different triglyceride (TG) compositions were labeled with [3H]cholesteryl oleoyl ether: LCT (triolein 100%), MCT:LCT (trioctanoin:triolein 50%:50%), MCT:LCT:FO (trioctanoin:triolein:triDHA 50%:40%:10%), and FO (triDHA 100%). Emulsions (0.4 mg TG/mouse) were injected into C57BL/6J mice, and blood clearance and tissue uptake of emulsion particles were determined. RESULTS: Large emulsion particles had 2- to 3-fold faster fractional catabolic rates (FCR) compared with small particles with the same TG content. There was 1.5- to 2.0 fold higher FCR of large FO-containing emulsions (FO and MCT:LCT:FO) compared with large LCT and MCT:LCT emulsions, whereas effects of FO on FCR in small emulsions were not observed. Large FO-containing emulsions were taken up more by adipose tissue compared with small particles with concomitant decreases in hepatic uptake. Preinjection of heparin reduced heart and adipose uptakes of FO and MCT:LCT:FO emulsions with increased uptake by liver, suggesting a role of lipoprotein lipase in catabolism of FO-containing emulsions. CONCLUSIONS: In a mouse model, FO addition to large emulsions increased blood clearance and changed organ delivery. In contrast, there was no or little effect when particle size became smaller. We hypothesize that in humans, FO addition to lipid emulsions can help target emulsion delivery to certain extrahepatic tissues, a factor that may be of use for delivering specific fatty acids, or even drugs, to specific organs. PMID- 12549601 TI - 3-0 methylglucose absorption in vivo correlates with nutrient absorption and intestinal surface area in experimental short bowel syndrome. AB - BACKGROUND: Inert carbohydrate probes are widely used to study intestinal permeability and examine the passive uptake of markers. This study examined the use of quantifying 3-0 methylglucose (3-0 MG) absorption as a marker of intestinal surface area and active nutrient transport capability. METHODS: Using a rat model, varying degrees of short bowel syndrome (SBS) were induced: sham operation (intestinal transection only), 50% resection (R50), 75% resection (R75), and 90% resection (R90; n = 6 to 8 in each group). Animals were pair fed, and over days 5 and 6 postoperation, a balance study was done to quantify absorption of dietary fat, protein, and carbohydrate. On day 7, animals were gavaged with the test solution containing 3-0 MG, lactulose, and mannitol, followed by a 16-hour urine collection. Urine recovery of probes was quantified using high-performance liquid chromotography. Animals were then killed, and the gross and microscopic intestinal morphology was determined. RESULTS: As expected, increased resection resulted in reduced absorption of dietary nutrients and 3-0 MG, which reached significance in the R90 resected animals. 3-0 MG absorption was significantly correlated with intestinal surface area and the absorption of dietary protein and fat (p < .01 for all comparisons). Interestingly, 3-0 MG absorption was not significantly correlated with the absorption of dietary carbohydrate. CONCLUSIONS: 3-0 MG absorption is a useful marker of functional intestinal surface area and of the absorption of dietary fat and protein in experimental SBS. 3-0 MG may be a useful marker of nutrient absorptive capacity in patients with SBS; further study is indicated. PMID- 12549602 TI - Serum and hepatic vitamin E assessment in cirrhotics before transplantation. AB - BACKGROUND: Hepatic vitamin E may have a protective effect against hepatocyte injury; therefore, vitamin E replacement or supplementation may be beneficial in patients with cirrhosis. However, serum vitamin E may not correlate with hepatic vitamin E stores, making decisions regarding treatment difficult based on serum levels alone. The specific aims of this study were to determine hepatic concentrations of vitamin E and to determine whether serum levels of vitamin E correlate with hepatic vitamin E stores in cirrhotics. METHODS: A prospective study of cirrhotics undergoing orthotopic liver transplantation (OLT) was completed. Serum and hepatic levels of vitamin E were measured by high performance liquid chromatography. Statistical analysis was performed using rank sum tests and Spearman's rank correlation coefficient. RESULTS: Fifty cirrhotics (33 males, 17 females; mean age of 53 years) were studied. The control group (25 males, 25 females; mean age of 47 years) consisted of the liver donors. The median serum levels of vitamin E in controls and cirrhotics were 5.95 and 7.8 mg/L, respectively (p = .009). The median hepatic levels (0.10 mg/g) in the control and cirrhotic groups were similar (p = .037). There was a significant correlation between serum and hepatic vitamin E levels in cirrhotics (R = 0.335; p = .017). CONCLUSIONS: A positive correlation exists between serum and hepatic concentrations of vitamin E in cirrhotics, therefore making serum vitamin E levels a useful reference for treatment using exogenous vitamin E. PMID- 12549603 TI - Nutrition support in the critical care setting: current practice in canadian ICUs -opportunities for improvement? AB - BACKGROUND: The purpose of this project was to describe current nutrition support practice in the critical care setting and to identify interventions to target for quality improvement initiatives. METHODS: We conducted a cross-sectional national survey of dietitians working in intensive care units (ICUs) across Canada to document various aspects of nutrition support practice. RESULTS: Of the 79 dietitians sent study materials, 66 responded (83%). Sixteen of 66 sites (24.2%) reported the presence of a nutrition support team, and 35 of 66 (53%) used a standard enteral feeding protocol. Dietitians retrospectively abstracted data from charts of all patients in the ICU on April 18, 2001. Of 702 patients, 313 (44.6%) received enteral nutrition only, 50 (7.1%) received parenteral nutrition only, 60 (8.5%) received both, and 279 (39.7%) received no form of nutrition support. Enteral nutrition was initiated on 1.6 days (median) after admission to ICU; 10.7% of patients were initiated on day 1. Of those receiving any form of nutrition support, on average, patients received 58% of their prescribed amounts of calories and protein over the first 12 days in the ICU. Of all days on enteral feeds, patients received feeds into the small bowel on 381 of 2321 (16.4%) days. The mean head of the bed elevation for all patients was 30 degrees. Controlling for differences in patient characteristics, site factors contributing the most successful application of nutrition support included the amount of funded dietitians per ICU bed, size of ICU, and the fact that the ICU was located in an academic setting. CONCLUSIONS: A significant number of critically ill patients did not receive any form of nutrition support for the study period. Those that did receive nutrition support did not meet their prescribed energy or protein needs, especially earlier in the course of their illness. Significant opportunities to improve provision of nutrition support to critically ill patients exist. PMID- 12549604 TI - Procalcitonin and enteral nutrition tolerance in critically ill patients. AB - BACKGROUND: Serum procalcitonin concentrations have been reported to be elevated in patients with bacterial infection. Early enteral nutrition (EN) has been shown to decrease infections in trauma patients. The purpose of this study was to characterize procalcitonin and other serum proteins during EN of trauma patients based on EN tolerance and presence of infection. METHODS: Twenty traumatized patients received a high-protein enteral formulation within 5 days of injury. Serum for procalcitonin, C-reactive protein, and prealbumin was analyzed on days 1 and 7 of EN. The procalcitonin/prealbumin and C-reactive protein/prealbumin ratios were calculated the same days. Patients who were infected during the study were compared with those not infected, and enteral-tolerant patients were compared with enteral-intolerant patients using these measurements. RESULTS: In the 20 trauma patients, procalcitonin (10.35 +/- 27.87 versus 1.03 +/- 1.24 ng/mL, p < .001) and procalcitonin/prealbumin ratio (1.70 +/- 4.20 versus 0.18 +/ 0.28, p < .01) decreased significantly over the 7-day period of EN. In the 12 patients who had infection, procalcitonin (16.33 +/- 35.31 versus 1.37 +/- 1.41 ng/mL, p < .004) and procal- citonin/prealbumin ratio (2.74 +/- 5.31 versus 0.26 +/- 0.33, p < .01) decreased significantly over the 7-day period of enteral nutrition. There were no significant changes in the measurements for 8 patients without infection. In the 15 patients who were enteral-tolerant, procalcitonin (12.56 +/- 32.84 versus 1.07 +/- 1.23 ng/mL, p < .004) and procalcitonin/prealbumin ratio (2.03 +/- 4.93 versus 0.20 +/- 0.29, p < .01) decreased significantly. CONCLUSION: Procalcitonin serum concentrations decrease significantly during EN in enteral-tolerant, critically ill patients with infection. PMID- 12549605 TI - Noncompliance with body weight measurement in tertiary care teaching hospitals. AB - BACKGROUND: Body weight provides vital information for patient care; therefore, measurement at hospital admission should be standard practice. Our objective was to test compliance with this standard. METHODS: This was a study of 300 patients, aged > or = 18 years, admitted to general medicine and surgery services of 3 tertiary care teaching hospitals in Nashville, Chicago, and San Francisco. At 24 to 36 hours after admission, participants were queried as to whether they had been weighed, and if not, they were asked whether they had been questioned by nursing personnel about their weight. Subjects were then weighed by research personnel using identical protocol at all 3 institutions. Any admission body weight documented by nursing was noted. RESULTS: Compliance was similar at all 3 institutions, with only 197 (65.7%) of patients reporting being weighed. There were 213 (71.0%) patients who had a weight documented in the nursing record. Of those who had not been weighed, 69 (67.0%) indicated that they had been queried about their weight. Comparison of documented weights in the nursing records with those measured by research personnel revealed that 55 (25.9%) differed by > or = 5 pounds (2.27 kg). Those who had a documented weight in the nursing record but were not weighed by nursing personnel were also more likely to deviate from the weight measured by research personnel by > or = 5 pounds (2.27 kg) in comparison with those who had been weighed by nursing personnel (42.8% versus 21.8%, respectively, p < .0147). CONCLUSION: Overall compliance with weight measurement is poor. Recorded weights are often inaccurate. PMID- 12549607 TI - The chicken soup paradigm and nutrition support: rethinking terminology. PMID- 12549606 TI - Pulmonary microcrystalline cellulose deposition from intravenous injection of oral medication in a patient receiving parenteral nutrition. AB - A 50-year-old man who had been dependent on home parenteral nutrition (HPN) for 24 years presented with shortness of breath. A computed tomography scan of the lungs revealed a diffuse micronodular parenchymal infiltrate. On bronchoscopy, a crystalloid material was identified. This organic material was determined to be consistent with codeine. The patient had been injecting codeine into his intravenous catheter. PMID- 12549608 TI - The compounding controversy: what veterinarians should know to protect themselves and their patients. PMID- 12549609 TI - Transdermal therapeutics. PMID- 12549610 TI - Clinicopathological, ultrasonographic, and histopathological findings of superficial necrolytic dermatitis with hepatopathy in a cat. AB - This report describes the antemortem diagnosis and antemortem and postmortem findings of superficial necrolytic dermatitis with hepatopathy (i.e., hepatocutaneous syndrome) in a cat. A 5-year-old Maine coon was evaluated because of a history of pruritic alopecia and liver enzyme elevations. Abdominal ultrasonography revealed a reticular pattern to the hepatic parenchyma. Histopathological findings of the liver were nodular regeneration with bands of vacuolated hepatocytes and bile duct hyperplasia, characteristic of the hepatopathy frequently associated with superficial necrolytic dermatitis. Skin histopathology revealed multifocal parakeratosis, midepidermal spongiosis, and basal cell hyperplasia consistent with superficial necrolytic dermatitis. PMID- 12549611 TI - The effects of four acidifying sprays, vinegar, and water on canine cutaneous pH levels. AB - This study determined the extent and duration of cutaneous acidification caused by a single application of four acidifying sprays, vinegar, and water. Multivariate repeated measures analysis of variance revealed a significant difference between the six sprays (F = 15.3; P < or = 0.001). Linear contrast tests showed that the effects of the acidifying sprays were significantly different from vinegar and water (F = 6.0; P < or = 0.001), and vinegar was significantly different from water (F = 13.8; P < or = 0.001). The acidifying sprays decreased cutaneous pH to < 6.0 for a mean range of 50 to 65 hours, while vinegar did so for a mean of 12 hours. PMID- 12549612 TI - Traumatic body wall herniation in 36 dogs and cats. AB - Traumatic body wall hernias (TBWH) are serious sequelae to traumatic injury in dogs and cats. During the study period, 26 dogs and 10 cats with surgically managed TBWH were identified. Five cases (four dogs, one cat) did not have their hernias identified during the first 24 hours of hospitalization. Bite wounds were the most common cause of TBWH, accounting for 54% of canine and 40% of feline hernias. Twelve cases (nine dogs, three cats) had serious intra-abdominal injuries diagnosed in addition to their TBWH. Seventy-three percent of dogs and 80% of cats survived to hospital discharge. In addition, the authors report the occurrence of a unique cause of herniation, termed an autopenetrating hernia. PMID- 12549613 TI - Gastric outflow obstruction after ingestion of wood glue in a dog. AB - A 2-year-old, male, mixed-breed dog presented with a 12-day history of vomiting, depression, and weight loss after ingestion of industrial-strength wood glue containing diphenylmethane diisocyanate as its active ingredient. A diagnosis of gastric foreign body was made from survey abdominal radiographs. A large aggregate of solidified wood glue was surgically removed, and the dog recovered uneventfully. Fourteen other cases have been reported to the Animal Poison Control Center at the American Society for the Prevention of Cruelty to Animals (ASPCA). Eight of those 14 cases required surgical intervention. All cases recovered completely. PMID- 12549614 TI - Prevalence of enteric pathogens in dogs of north-central Colorado. AB - To evaluate the prevalence of enteric pathogens in dogs of north-central Colorado, fecal samples were obtained from client-owned dogs presented to the Veterinary Teaching Hospital at Colorado State University for evaluation of acute small-bowel, large-bowel, or mixed-bowel diarrhea (n=71) and from age-matched, client-owned, healthy dogs (n=59). Infectious agents potentially associated with gastrointestinal disease were detected in 34 of 130 (26.1%) fecal samples. Agents with zoonotic potential were detected in feces from 21 (16.2%) of 130 dogs and included Giardia spp. (5.4%), Cryptosporidium parvum (3.8%), Toxocara canis (3.1%), Salmonella spp. (2.3%), Ancylostoma caninum (0.8%), and Campylobacter jejuni (0.8%). Positive test results occurred in dogs with or without gastrointestinal signs of disease. Dogs, particularly those in homes of immunocompromised humans, should be evaluated for enteric zoonotic agents. PMID- 12549616 TI - Juvenile hyperthyroidism in a cat. AB - An 8-month-old, male domestic shorthaired cat presented for chronic weight loss, intermittent dyspnea, chronic diarrhea, hyperactivity, and weakness. The cat had a palpable thyroid nodule and increased serum total thyroxine and 3,5,3' triiodothyronine levels. The cat was diagnosed with hyperthyroidism, and a unilateral thyroidectomy was performed followed by radioactive iodine at a later date. The clinical signs resolved following radioactive iodine, and the cat subsequently developed clinical hypothyroidism. PMID- 12549615 TI - Ultrasonographic signs of intestinal intussusception associated with acute enteritis or gastroenteritis in 19 young dogs. AB - The ultrasonographic pattern of intestinal intussusception was studied in 19 young dogs with acute enteritis or gastroenteritis. The intussusception was observed to be a target-like mass consisting of a hyperechoic or anechoic center surrounded by multiple hyperechoic and hypoechoic concentric rings in transverse sections. In longitudinal sections, the intussusception appeared as multiple hyperechoic and hypoechoic parallel lines, as a tumor-like or pseudokidney mass, and as a trident-like configuration. Ultrasonography may provide a sensitive, specific, and accurate method of diagnosing intestinal intussusception in young dogs. PMID- 12549617 TI - Clinicopathological evidence of pseudomyxoma peritonei in a dog with intestinal mucinous adenocarcinoma. AB - An 11-year-old, intact female Pekingese dog was presented because of chronic vomiting. Clinical examination and diagnostic imaging suggested the presence of an intestinal mass. Laparotomy was performed, and a gelatinous effusion associated with a suspected jejunal neoplasm was found. Accumulation of gelatinous material was grossly evident in several abdominal organs and parietal peritoneum. Cytopathological smears from this material showed macrophages, reactive mesothelial cells, and spindle cells embedded in a mucinous basophilic background. After spontaneous death, necropsy and histopathology were performed and showed the presence of an invasive mucinous adenocarcinoma with visceral and peritoneal metastases. The clinicopathological findings of this case report closely resemble those reported in human cases of pseudomyxoma peritonei. PMID- 12549618 TI - Treatment of renal nephroblastoma in an adult dog. AB - An 8-year-old Labrador retriever was diagnosed with a unilateral malignant nephroblastoma and hypertrophic osteopathy. The histopathologically malignant tumor was confined to the renal capsule, but the sarcomatous component was anaplastic, resulting in its classification as a Stage I tumor with unfavorable histopathology. The dog was treated with unilateral nephrectomy, vincristine, and doxorubicin. This dog has remained disease free for >25 months. Reported treatments of renal nephroblastoma in the dog have not described disease-free intervals of >8 months. PMID- 12549619 TI - Extradural spinal, bone marrow, and renal nephroblastoma. AB - A 1-year-old, female intact Shetland sheepdog presented with acute onset of neurological signs. Physical examination revealed a large abdominal mass. Neurological examination revealed multifocal disease with neck pain, short strided forelimbs, and hind-limb paresis with loss of tail and anal tone. Blood work, imaging techniques, cytopathology, and histopathology led to a diagnosis of renal, bone-marrow, and extradural spinal nephroblastoma. This report documents potential clinical and pathological manifestations of canine nephroblastoma that have not been previously reported. PMID- 12549620 TI - Efficacy of cobalt-60 radiation therapy for the treatment of nasal cavity nonkeratinizing squamous cell carcinoma in the dog. AB - The purposes of this study were to evaluate the efficacy of cobalt-60 radiotherapy in the treatment of nonkeratinizing squamous cell carcinoma of the nasal cavity in dogs and to compare this treatment group to historical controls. Six dogs with histopathologically confirmed nasal cavity nonkeratinizing squamous cell carcinoma were treated with cobalt-60 radiotherapy to a total dose of either 63 Gy or 54 Gy. Overall survival times ranged from 30 days to 330 days, with a median survival time of 165 days. Nasal cavity nonkeratinizing squamous cell carcinoma in the dog is an aggressive tumor that responds poorly to radiotherapy. PMID- 12549621 TI - Intravascular lymphoma involving the central and peripheral nervous systems in a dog. AB - A 5-year-old, castrated male mixed-breed dog was presented for paraparesis, ataxia, hyperesthesia, and thrombocytopenia of 5 months' duration and recurrent seizures during the preceding 2 weeks. Multifocal neurological, ophthalmological, pulmonary, and cardiac diseases were identified. Magnetic resonance imaging and cerebrospinal fluid analysis supported a tentative diagnosis of neoplastic or inflammatory disease. A computed tomography-guided biopsy provided both cytopathological and histopathological evidence of intravascular lymphoma. The disease progressed despite chemotherapy with prednisone, L-asparginase, and vincristine. Postmortem histopathological examinations suggested intravascular lymphoma in the central and peripheral nervous systems as well as in multiple other organ systems. This is the first description of an antemortem diagnosis and treatment of intravascular lymphoma involving the central nervous system of a dog. PMID- 12549622 TI - Distraction osteogenesis for treatment of premature physeal closure and shortening of the third and fourth metatarsals of a dog. AB - A 4-month-old Siberian husky was presented for a history of lameness of the left hind limb. Physical and radiographic examination revealed a 1-cm shortening of metatarsals III and IV, with subsequent hyperextension of the digits on the left hind paw. A circular external skeletal fixator was used to accomplish distraction osteogenesis in both metatarsals over a period of 15 days. Treatment was successful in restoring appropriate length of the metatarsals and in resolution of the lameness. At 6 and 12 months after surgery, the owner reported that the dog was ambulating normally and had no complications related to the surgery. PMID- 12549623 TI - Constitutive production of parathyroid hormone-related protein (PTHrP) by fibroblasts derived from normal and pathological human breast tissue. AB - Parathyroid hormone-related protein (PTHrP) is expressed in the mammary gland and appears to be critical to the morphogenesis of this structure. PTHrP production in the breast is generally attributed to epithelial cells. Because the stromal component of the breast produces factors implicated in proliferation and differentiation of mammary epithelial tissue and tumors, the aim of this study was to investigate the PTHrP expression by mammary fibroblasts from breast cancer tumors and normal breast. PTHrP antibodies labeled intralobular fibroblasts in normal breast and stromal fibroblasts that surround tumor cells. PTHrP was constitutively produced by the cultured mammary fibroblasts, independent of serum stimulation. Normal (15.83 +/- 1.72 fmol/10(6) cells) and pathological breast fibroblasts (19.87 +/- 5.76) secreted similar amounts of PTHrP. PTH/PTHrP receptor mRNA was detected by RT-PCR in all the samples tested. Fibroblasts from normal breast were both PTH and PTHrP-cAMP responsive (453 +/- 133% and 513 +/- 133%, respectively, from basal stimulation), whereas pathological breast fibroblasts were minimally PTHrP-cAMP responsive (183 +/- 36%). The production of other fibroblastic factors implicated in tumor growth and invasiveness was also examined. Interleukin-6 (IL-6), tumor necrosis factor-alpha (INF-alpha), and pro matrix metalloproteinase (MMP)-1 were not affected by the status of the tissue. In contrast, increased levels of pro-MMP-2 were produced in fibroblasts that originated from pathological (290 +/- 62 ng/10(6) cells) samples compared with those from normal donors (125 +/- 41 ng/10(6) cells). PTHrP production was correlated with TNF-alpha and pro-MMP-2 production. However, inhibition with specific neutralizing antibodies against TNF-alpha or PTHrP, or with a PTHrP antagonist, showed that these factors did not regulate each other. In conclusion, breast fibroblasts are constitutive PTHrP-producing cells with the potential for autocrine signaling through the PTH/PTHrP receptor. PMID- 12549624 TI - Embryonal mass and hormone-associated effects of pregnancy inducing a differential growth of four murine tumors. AB - A differential effect of pregnancy on the growth of subcutaneous implants of four murine tumors has been observed. Two tumors lacking receptors for progesterone and estrogen [methylcholanthrene-induced fibrosarcoma (MC-C) and spontaneous lymphoid leukemia (LB)] exhibited slow kinetics throughout the course of pregnancy, although inhibition was stronger beyond day 10. On the other hand, one of two tumors bearing receptors for progesterone and estrogen [medroxyprogesterone (MPA)-induced mammary adenocarcinoma (C7HI)] exhibited three phases: up to days 8-10 of gestation the tumor grew faster than in virgins, between days 8-10 and 15 it reached a plateau, and beyond day 15 a sharp reduction in tumor mass was observed. The other tumor [mouse mammary tumor virus (MMTV)-induced mammary carcinoma(T2280)] behaved as a typical pregnancy-dependent tumor (i.e., it grew in pregnant but not in virgin mice, regressed soon after delivery, and reassumed its growth at the middle of a second round of pregnancy). Neither MPA nor estrogen affected MC-C and LB tumor growth. On the other hand, MPA-treated mice enhanced C7HI tumor and reciprocally C7HI tumor-bearing mice treated with estrogen strongly inhibited tumor growth. As for T2280, neither MPA nor estrogen alone could promote tumor growth and, in consequence, no tumor developed. However, when MPA plus estrogen was administered in a schedule simulating the successive appearance of these hormones in pregnancy, T2280 grew even faster than in pregnant mice. When the four tumors were implanted in mice bearing grafts of embryonal tissues (teratomas), all of them were inhibited. This antitumor effect was similar to that observed in pregnancy when tumors unresponsive to progesterone and estrogen were tested. On the other hand, with tumors bearing progesterone and estrogen receptors, differences in tumor growth were detected in pregnant and teratoma-bearing mice. This suggested the existence during pregnancy of two factors potentially acting on tumor growth. First, a progesterone and estrogen-mediated hormonal component, which would exert either inhibitory or stimulatory effects only evidenced with tumors bearing hormonal receptors. Secondly, an antitumor effect proportional to the growing embryonal mass, inhibiting all tumors independently of their origin or hormone responsiveness. This antitumor effect could be attributed to a beat-resistant serum factor (1,000-1,200 Da molecular weight) presumably associated with the pathway of the arachidonic acid metabolism. The interplay between the hormonal component and the serum factor associated with embryonal mass could account for some of the largely heterogeneous and otherwise unexplained effects of pregnancy on tumor growth reported in the literature and illustrated by the four tumors studied here. PMID- 12549625 TI - Analysis of differentially expressed genes in hepatocellular carcinoma with hepatitis C virus by suppression subtractive hybridization. AB - Hepatitis C virus (HCV) infection is associated with pathogenesis of hepatocellular carcinoma (HCC). We carried out suppression subtractive hybridization to identify variable expression of genes linked to HCC with HCV infection. RNA from both tumorous (tester) and nontumorous (driver) liver tissues was isolated. The cDNA clones were subjected to MegaBACE PCR sequencing to identify those that hybridized to the subtracted library with preference. Nucleic acid sequences generated were searched against the human UniGene database. Among 576 clones screened in the tumorous liver tissue, we identified 30 genes and 28 expressed sequence tags (ESTs). Among 30 genes detected, 23 were with known functions and 7 with unknown functions. The known genes identified had diversified functions and could be divided into 10 functional categories. Twenty percent of these genes were previously known to be tumor related and those most frequently appearing were haptoglobin alpha(2FS)-beta precursor, haptoglobin related protein, and alpha-2-macroglobulin. Four out of 30 known genes (immunoglobulin lambda light chain, kappa immunoglobulin, spliceosomal protein, and X-ray repair cross-complementing protein) were related to chromosome translocation and nucleotide repair. These four genes may contribute to carcinogenesis caused by DNA-damaged agents and to the efficiency of anticancer therapy. The genes with unknown function, which were most frequently detected, were PRO2760 and PRO2955; both encode proteins that express in fetal liver. Twenty-one known and six novel genes were discovered in the nontumorous liver tissue. Apparently, these 27 genes were lost in the tumorous liver tissues. Therefore, using suppression subtractive hybridization, we have identified a number of genes associated with HCC with HCV infection. Most of these genes have not been reported in HCC. Further characterization of these differentially expressed known and unknown genes will provide useful information in understanding the genes responsible for the development of HCC. PMID- 12549626 TI - A sensitive bioassay for measuring blood levels of 12-O-tetradecanoylphorbol-13 acetate (TPA) in patients: preliminary pharmacokinetic studies. AB - 12-O-Tetradecanoylphorbol-13-acetate (TPA) is a potent stimulator of differentiation in myelocytic leukemia cells, and it has been shown to have activity in patients with acute myelocytic leukemia. Because attempts to develop a suitable mass spectrometry assay for TPA were unsuccessful (because of the lack of sufficient sensitivity), we developed a novel and highly sensitive blood level bioassay for TPA that measures ethyl acetate-extractable differentiating activity in blood. Differentiating activity in ethyl acetate extracts of blood was measured in HL-60 cells by measuring the formation of adherent cells. The sensitivity of the assay was approximately 0.1 ng TPA/ml blood. The assay for TPA has a high degree of specificity and does not measure deesterifed potential metabolites (phorbol, phorbol-13-acetate, or phorbol-12-myristate), and the presence of GM-CSF, G-CSF, interferon-alpha, or interferon-gamma does not interfere with the assay. Blood levels of TPA as measured by the bioassay immediately after an IV infusion of TPA (0.125 mg/m2; approximately 0.25 mg per patient) and 1 and 3 h later were 1.75 +/- 0.55, 0.93 +/- 0.54, and 0.69 +/- 0.42 ng/ml, respectively (mean +/- SD from eight infusions in five patients). Terminal half-lives were determined in a few patients where TPA blood levels were measured at multiple time intervals after the TPA infusion. In these patients, the terminal half-life was 11.1 +/- 3.9 h (from five infusions in four patients). To the best of our knowledge, this is the first analytical method for the measurement of TPA. PMID- 12549627 TI - Antitumor polycyclic acridines. Part 12. Physical and biological properties of 8,13-diethyl-6-methylquino[4,3,2-kl]acridinium iodide: a lead compound in anticancer drug design. AB - The biophysical and biological characterization of 8,13-diethyl-6 methylquino[4,3,2-k]lacridinium iodide (6) is reported. The compound binds to DNA, as measured by UV, fluorescence, and circular dichroism studies, and stabilizes the double helix and higher order DNA structures (DNA triplexes and quadruplexes) against thermal denaturation. Unlike many DNA ligands, (6) shows no specificity for binding to specific base pair combinations and does not inhibit topoisomerase I (topo I) or topo II activity. Furthermore, the biological fingerprint elicited by (6) in in vitro evaluations does not compare with clinical agents of the topo II inhibition class. The compound provokes cell cycle arrest in response to DNA damage and the biological sequelae are dependent on the p53 status of the cell line. DNA damage by (6) upregulates p53 and p21(CIP/WAF1) proteins. The unusual structure of (6) and its ease of synthesis in a "one-pot" reaction are features that are being exploited in the design and development of a new series of G-quadruplex stabilizing telomerase inhibitors. However, although the second-generation compounds that resulted from (6) present strong telomerase inhibition, (6) in itself presents yet a different mode of action, with a strong preference for triplex DNA, sequences often found in a number of genes. PMID- 12549628 TI - Penile lipomas, cause of sexual dysfunction. AB - We report 2 cases of penile lipomas. Although the simultaneous sexual dysfunction proved to be phsychogenic their surgical excision improved their sexual life. PMID- 12549629 TI - Delayed suture intravesical migration as a complication of a Stamey endoscopic bladder neck suspension. AB - We report our experience with two cases of late migration of the suture and bolster occurring 2 years after a modified Stamey endoscopic bladder neck suspension. Delayed migration of the suture and bolster after an endoscopic bladder neck suspension across tissue planes, with subsequent erosion into the bladder, is uncommon. Recurrent urinary tract infection and mild suprapubic discomfort were the only symptoms. Cystoscopy was the only helpful diagnostic tool and should be considered early in the evaluation of this kind of patients. The mechanism of migration of the cuff and the operative technique are discussed. PMID- 12549630 TI - Recurrent adrenal pheochromocytoma: a case report. AB - We report an unusual case of benign recurrent pheochromocytoma that developed at the same site 8 years following initial treatment. This tumor was managed successfully through the 11th rib bed using the transperitoneal extrapleural approach. PMID- 12549631 TI - Treatment of indinavir sulfate induced urolithiasis in HIV-positive patients. AB - Indinavir sulfate is a protease inhibitor used of the treatment of primary HIV infection either as monotherapy or as part of antiretroviral treatment schemes. Approximately 10% of all patients develop urolithiasis with radiolucent stones consisting of indinavir. We present our results of the treatment in 11 HIV positive patients (9 men, 2 women), who developed Indinavir lithiasis after 5-8 months of antiretroviral therapy. Following the initial procedures (spasmoanalgetic drugs, ureteroscopy, double J-stent or nephrostomy), the patients were further treated by increasing diuresis and urinary acidification. All the patients responded well to the treatment, the obstruction was releieved and their renal function was restored to normal. PMID- 12549632 TI - Paradoxical incontinence caused by "invisible dysplastic" kidneys: role of laparoscopy. AB - A 35-year old female presented with paradoxical urinary incontinence. Examination under general anesthesia did not reveal any abnormality and cystoscopy showed absent right hemitrigone. Radiological and other renal imaging modalities revealed absent right kidney and normal functioning left kidney. Simultaneous localization and removal of small dysplastic right kidney was carried out by transperitoneal laparoscopy. Role of laparoscopy is emphasized in patients presenting with paradoxical urinary incontinence with no "visible" kidney/ureteric orifice on conventional evaluation. PMID- 12549633 TI - Cystic nephroma: a rare clinical entity. AB - We report a rare benign renal tumor, the cystic nephroma (CN) in a 35-year old man. CN should be considered in the differential diagnosis when clinical and radiological examinations reveals a multilocular cystic renal mass, but histopathological examination is the unique diagnostic mean. PMID- 12549634 TI - Haemophilus influenzae acute pyelonephritis in the elderly. AB - Haemophilus influenzae has rarely been implicated as the causative agent of urinary tract infections in adults. We report a case of H. influenzae pyelonephritis in a 78-year old man with benign prostatic hyperplasia and discuss the possible underestimation of the true incidence of H. influenzae in adult urinary tract infections. PMID- 12549635 TI - Speedy elimination of ureterolithiasis in lower part of ureters with the alpha 1 blocker--Tamsulosin. AB - BACKGROUND: Alpha-I blockers decrease the tension and release the spasm of smooth muscles and thus lessen the obstruction and irritation symptoms in the lower urinary tract (LUTS). They make a faster passing of calculi from the terminal part of the ureters possible. OBJECTIVES: The goal of this study was to objectively assess the improvement of difficulties caused by obstructions in ureterolithiasis localized in the lower part of the ureters of 104 randomly chosen patients (pts.) in a double-blind study. METHODS: During a period of 2 and half years (June 1999-January 2002) 104 pts. suffering from ureterolithiasis of the lower urinary tract were treated and observed. Patients were divided into two groups: A (n:53; later only 51 were evaluated) which was subjected to standard treatment and group B (n:51) where the standard treatment was supplemented by the alpha-1 blocker. As alpha-1 blocker one capsule of Tamsulosin/OMNIC 0.4/ was administered daily. RESULTS: With alpha-1 blocker, we have registered a more speedy passing of calculi from the terminal parts of ureters in 17.6% of pts. Recurrence of renal colics was less frequent and occurred in one of eight pts. as compared with group A (without the alpha-1 blocker) where a recurrence of the renal colic was observed in about every fifth pts. In group A (n:51), 62.8% of the pts. passed the calculi, whereas in group B (n:51), where standard treatment was supplemented by the administration of the alpha-1 blocker Tamsulosin, this percentage increased to 80.4%. CONCLUSION: The treatment by alpha-1 blockers considerably decreased not only LUTS but also helped to accelerate the passing of minor calculi from the terminal parts of the ureters of 80.4% of pts. It seems that alpha-1 blockers potentiate the spasmoanalgetic action of drugs used in standard methods of treatment. PMID- 12549636 TI - Management of iatrogenic ureteric injuries associated with gynecological surgery. AB - OBJECTIVE: This retrospective study defines the presentation and management of iatrogenic ureteric injuries consequent upon gynecological surgery in a teaching hospital in Punjab, Pakistan. PATIENTS AND METHODS: 18 patients with median age 35 years (range 18-80 years) with iatrogenic ureteric injuries associated with gynecological surgery were referred to the department of urology at Nishtar Hospital Multan Pakistan. Main presenting symptoms were urinary incontinence, loin pain and anuria. Median time since injury and presentation was 3 weeks (range 1 day to 7 years). In 16 (88%) patients injury resulted from abdominal hysterectomy. Other causes included ovarian cystectomy (one patient) and vaginal hysterectomy (one patient). 11 (61%) patients had ureterovaginal fistula, 5 (28%) patients had complete unilateral ureteric obstruction and 2 (11%) patients had bilateral ureteric obstruction and anuria. In 11 patients with ureterovaginal fistula ureteroneocystostomy was performed. In five patients with unilateral ureteric obstruction, one had end to end anastomosis of ureter, three had ureteroneocystostomy only and one had ureteroneocystostomy and psoas hitch done. Two had anuria secondary to bilateral ureteric obstruction. In one of these patients Boari flap and ureteroneocystostomy was carried out. The second patient had deligation of catgut sutures on ipsilateral side and ureteroneocystostomy on the contra-lateral side. RESULTS: In 17 patients no major complication occurred. One patient who had deligation of catgut sutures, the distal ureter sloughed and re-exploration and ureteroneocystostomy was performed. Renal salvage was achieved in all cases. CONCLUSION: Open surgical procedures for repair of iatrogenic ureteric injuries are associated with good outcome. Strategies to prevent these injuries include adequate surgical training and meticulous surgical techniques. PMID- 12549637 TI - Partial outlet obstruction of the rat bladder induces a stimulatory response on proliferation of the bladder smooth muscle cells. AB - Outlet obstruction of the rat bladder induces hypertrophy/hyperplasia characterized by increases in bladder mass, smooth muscle content, and collagen deposition. In order to understand the mechanism of the outlet obstruction induced hypertrophy and hyperplasia, we first determined the temporal pattern of changes in bladder mass after inducing the outlet obstruction. Histological analysis revealed that the smooth muscle cells with hypertrophy and hyperplasia, fibroblasts and connective tissue were increased in a time-dependent manner, corresponding to the temporal pattern observed in the changes in bladder mass, although the phase of changes in these tissue components was somewhat different. In order to further determine whether any proliferation-stimulatory factors were released from the bladder with obstruction in correspondence with increased bladder mass, soluble fractions were prepared from the bladders with outlet obstruction for 3-30 weeks, and their effects on proliferation of smooth muscle cells were examined. The soluble fractions prepared from the bladders at 3 to 14 weeks after obstruction slightly but significantly facilitated the proliferation of cultured smooth muscle cells, while the soluble fractions released after 20 weeks rather suppressed the proliferation. These results suggest that the initial increase in bladder mass might be in part due to the facilitated proliferation of smooth muscle cells of the bladder body induced by growth factors released into the soluble fractions, and that hypertrophy might then play a role in the increased bladder mass at later phases. PMID- 12549638 TI - Long-term safety of extended-release oxybutynin chloride in a community-dwelling population of participants with overactive bladder: a one-year study. AB - In this multicenter, open-label study of extended- and immediate-release oxybutynin chloride, community-dwelling participants were studied for up to 12 months to evaluate the long-term safety profile of extended-release oxybutynin. Quality-of-life assessments designed to measure the impact of incontinence and evaluate treatment outcome were used to study subjective improvement. A total of 904 women and 163 men (mean age 64 years, range 29-91 years) were enrolled. The majority of discontinuations were in the first 3 months (25.5%); of those who continued after 3 months, 62% remained on extended-release oxybutynin chloride for one year. The majority of discontinuations were for adverse events; dry mouth was the most frequently cited event leading to discontinuation (8.4%). Significant improvements were seen in QOL measures. Long-term therapy with extended-release oxybutynin chloride was generally well tolerated and effective, improving quality of life significantly in participants with overactive bladder over 3-12 months of therapy. PMID- 12549639 TI - Eye metastasis form urothelial tumours. AB - Obit is an infrequent site of metastasis from urothelial tumours. To the best of our knowledge there are less than ten case reports of orbital or choroidal metastases from urothelial tumours reported in English medical literature. On the basis of review of these reports (Table 1) and experience in managing two more cases, it is emphasized that non-specific eye symptoms in patients of urothelial tumours need thorough ophthalmological and radiological evaluation. Herein, we present case descriptions, along with pathophsiology; and review of literature to highlight the need for early diagnosis and prompt management of this rare clinical event. PMID- 12549640 TI - Idiopathic localised bladder amyloidosis: rare cause of haematuria. AB - Idiopathic Localised Bladder Amyloidosis is a rare cause of haematuria and urinary tract symptoms. A review of the literature highlights the varied presentations and the appropriate investigations for this condition, with emphasise on the exclusion of a secondary cause. In addition, the range of treatment options is fully discussed. Our report on a 65-year old gentleman illustrates that a high index of suspicion is required for its diagnosis. Furthermore, conservative management can be an effective strategy in selected patients. PMID- 12549641 TI - The significance of random bladder biopsies in superficial bladder cancer. AB - INTRODUCTION: Today, there is no consensus about taking random bladder biopsies during transurethral resection of superficial bladder tumors for staging and to determine the urothelial abnormalities like dysplasia and carcinoma in situ. The aim of our study was to evaluate the results and indications of random bladder biopsies for primary superficial bladder cancer. PATIENTS AND METHODS: Random bladder biopsies were taken from 84 patients with primary superficial bladder cancer after transurethral resection. 40 patients had Ta and 44 had T1 tumor. The random biopsies were taken from right and left bladder walls, anterior and posterior walls, dome, trigone and prostatic urethra. The incidence of urothelial abnormalities were evaluated according to the stage and grade of the tumor. RESULTS: None of the patients had carcinoma in situ or dysplasia with Ta tumor. In T1 group, 4 patients (9.1%) had carcinoma in situ and 3 patients (6.8%) had dysplasia. There was a statistically significant difference with regard to urothelial abnormalities between groups Ta and T1. The same difference was also seen between low and high grade tumors. CONCLUSION: In our study, only 7/84 (8.3%) of patients with primary superficial bladder cancer had urothelial abnormalities like carcinoma in situ or dysplasia. All of these pathologies were seen in T1 tumors. According to our results, we believe that random biopsies are not useful in superficial bladder cancers to detect urothelial abnormalities and also do not help for the planning of further treatment. PMID- 12549643 TI - Malignant anterior urethral strictures: a rare complication of transurethral resection of malignant prostate. AB - A 65-year old male presented with obstructive voiding symptoms and recurrent hematuria following transurethral resection of malignant prostates. Following an unsuccessful attempt at catheterization, urethroscopy revealed narrowing of anterior urethra secondary to multiple nodular lesions. Biopsy of lesion showed adenocarcinoma positive for prostate specific antigen. This rare presentation has been reported only twice in past. Clinical presentation, pathophysiology and brief review of literature are discussed. PMID- 12549642 TI - Immunopathological prognostic and predictive factors in prostate cancer. AB - Prostate cancer is the leading male malignancy in the Western world. Patients with prostate cancer have an unpredictable clinical course, as three biologically different types of tumor exist. This review summarises some of the recent progress made in understanding the biology of prostate cancer with special reference to the prognostic and predictive role of immunohistochemical markers. The prognostic value of established prognostic variables is also discussed. PMID- 12549645 TI - The effect of nitric oxide in testicular ischemia-reperfusion injury. AB - This experiment was carried out to investigate the effect of endogenous nitric oxide (NO) on the ischemia-reperfusion injury of testis. Testicular ischemia was achieved by twisting the right testis and spermatic cord 1080 counter-clockwise for 30 minutes and reperfusion was allowed for 30 minutes after detorsion of 33 rats. Animals were treated with normal saline in controls just before detorsion, NG-nitro-L-arginine methyl ester (L-NAME), and L-arginine (L-arg) in others. The tissue damage was evaluated with light microscopy, malondialdehyde (MDA) level in tissue, and the blood flow measurement using 133xenon (Xe) clearence technique. MDA indicator of reperfusion injury increased 25% after detorsion when only normal saline was given, L-NAME further increased MDA, L-arginin decreased MDA to control level. CONCLUSION: L-arginin infusion during the detorsion reduced the reperfusion injury of testis and improved the testicular blood flow after the detorsion. PMID- 12549644 TI - Effects of combined androgen blockade on bone metabolism and density in men with locally advanced prostate cancer. AB - OBJECTIVE: To investigate whether combined androgen blockade (CAB) produces any adverse effects on bone metabolism and mineral density in patients with locally advanced prostate cancer. MATERIALS AND METHODS: The study group consisted of 17 stage T4 prostate cancer patients treated with CAB and had no evidence of bone metastasis on bone scintigraphy. The mean duration of CAB and final total prostate specific antigen (PSA) level at the time of study were found at 28.5 +/- 15.9 (6-58) months and 0.39 +/- 0.5 (0.1-2) ngml, respectively. Twenty age and socioeconomically matched benign prostate hyperplasia (BPH) patients were taken as the control group. Both groups were compared with regard to lumbar bone mineral density (LBD), femur bone mineral density (FBD) and serum parameters of bone metabolism namely calcium (Ca), phosphate (P), magnesium (Mg) and alkaline phosphatase (ALP). Bone mineral density was measured with dual energy x-ray absorptiometry. RESULTS: The mean FBD, LBD and serum Ca, P, Mg and ALP measurement of the patients treated with CAB were 0.85 +/- 0.1 g/cm2, 1.16 +/- 0.2 g/cm2, 9.1 +/- 0.3 mg/dl, 3.6 +/- 0.6 mg/dl, 1.95 +/- 0.14 mg/dl, 187.5 +/- 61 mg/dl, respectively. No significant difference was found between patients subjected to CAB and the age matched controls in any of the studied parameters namely age, FBD, LBD, Ca, Mg and ALP except serum phosphate. Serum phosphate levels were significantly (p = 0.001) higher in patients treated with CAB suggesting a minor effect of CAB on bone metabolism. CONCLUSION: No convincing evidence was found about the detrimental effect of CAB on bone mineral density and metabolism in a highly selected group of patients with advanced prostate cancer without bone metastases. PMID- 12549646 TI - Adenomatous hyperplasia of the rete testis: report of two cases. AB - We present two adenomatous hyperplasia of the rete testis (AHRT) cases. One of them was a 67-year old patient with prostatic adenocarcinoma and the other was a 38-year old patient with undescended testis. AHRT is a rarely seen lesion and usually detected as incidental microscopic finding. It may be confused with malignancy and related to testicular atrophy and hormonal imbalance. The treatment of choice is complete excision. PMID- 12549648 TI - Idiopathic penile mondors' disease: a case report. AB - A 34-year old man was admitted to us with a mild painful cord-like induration on the left dorsal side of the penis extending from corona of the glans to the base of the penis. The patient was treated with anti-inflammatory and anticoagulan agents. Cord-like-induration resolved and recanalized in 4th week, and venous flow was detected in Doppler examination. PMID- 12549647 TI - Bellini duct carcinoma: review of diagnosis and management. AB - AIM: To review the world literature on the histogenesis, diagnosis and management of Bellini duct carcinoma and to suggest a possible clinical algorithm to assist in their identification and appropriate therapeutic strategies. METHODS: A medline review of all reported cases of Bellini duct carcinoma was carried out from the Pubmed using the keywords collecting duct carcinoma, Bellini duct carcinoma, medullary renal carcinoma and renal cancers. RESULTS: 40 worldwide reported cases of Bellini duct carcinoma were found. We analyzed their salient pathological, diagnostic and management strategies. A simplistic approach and a clinical algorithm has been suggested to enable their rapid identification, diagnosis and management. PMID- 12549649 TI - Primary urethral tuberculosis masquerading as a urethral caruncle: a diagnostic curiosity! AB - Primary urethral tuberculosis associated with a caruncle is an extremely rare entity and ours is the second such case to be reported. A middle-aged woman presented with symptoms of frequency dysuria syndrome for the last 2 years. Local examination and cystoscopy revealed localized parurethral induration, tenderness and a urethral caruncle with chronic obliterative urethritis. Transvaginal ultrasound revealed a solid lesion arising from the posterior urethral wall. Excision of the caruncle and transurethral resection of the paraurethral mass lesion revealed chronic granulomatous inflammation with chronic urethritis and fibrocollagenous tissue. The literature regarding the genesis of urethral caruncle, the underlying conditions, its differential diagnosis and management has been reviewed. PMID- 12549650 TI - Peyronie's disease: an unusual presentation with butterfly shaped calcified plaques. AB - Peyronie's disease is characterized by the formation of a fibrotic plaque in the tunica albugenia of the penis. Two patients with calcified butterfly shaped plaques involving the ventrolateral aspect of the penis are described. These patients were managed by plaque excision and dermal grafting with excellent post operative results. PMID- 12549651 TI - Penis agenesis associated with mental retardation: a case report. AB - Penile agenesis is an extremely rare anomaly, with a reported incidence ranging from 1 in 10 million to in 3 million births. There are only about 80 published cases. This report first describe of penile agenesis associated with mental retardation. PMID- 12549652 TI - Automutilation of the penis performed by the kitchen's knife. AB - Automutilation of the penis performed by the kitchen's knife is described. Surgical techniques, the most important goals of therapy as well as psychiatric aspects of the automutilation are discussed. PMID- 12549653 TI - Comparison of nocturnal penile tumescence monitoring and cavernosal smooth muscle content in patients with erectile dysfunction. AB - PURPOSE: Nocturnal penile tumescence monitoring was compared to cavernosal smooth muscle content in 48 cases of erectile dysfunction. MATERIALS AND METHODS: Pre operatively nocturnal penile tumescence rigidity (NPTR) testing, colour Doppler sonography and if needed pharmaco cavernosometry-cavernosography were evaluated in 48 impotent patients before surgical intervention. The 40 patients whom all those diagnostic tools were abnormal constituted the first group. In the remaining 8 patients, which constitutes the second group, NPTR testing were normal but the other tests were abnormal. 10 potent patients with congenital penile curvature constituted the third group. Cavernous biopsies were obtained during the surgery and biopsies stained immunohistochemically to quantify smooth muscle cells (SMC) by anti-desmin and anti-SMA. RESULTS: We observed statistical significant difference of corporeal SMC content with regard to first Vs second group and first Vs third group (p < 0.05). However we did not observe statistically significant difference with regard to second vs third group (p > 0.05). CONCLUSION: NPTR testing appears to correlate well with corporeal SMC, which is the key structures of erection. We think that with taking into the consideration of its specific reservations, NPTR testing is still one of the best non-invasive tool in the differential diagnosis of erectile dysfunction. PMID- 12549654 TI - Is midazolam effective as an antioxidant in preventing reperfusion injury in rat kidney? AB - This experimental study was designed to investigate whether midazolam has antioxidant effects in reperfused rat kidneys following ischemia. Twenty Wistar Albino rats were included in the study. Rats were anesthetized with the mixture of ketamine 90 mg/kg and xylazine 10 mg/kg administered intraperitoneally. Following anesthesia, the rats were divided into two groups. The first group was considered as the control group, whereas the second group received additional midazolam 3.5 mg/kg intraperitoneally. The left kidney was approached via a transabdominal incision and the left renal artery was dissected. Left renal ischemia was created by clamping the left renal artery for 45 minutes. Following the ischemia period, the kidney was reperfused for one hour. Both kidneys were then removed. Half of the left kidneys were immediately immersed in liquid nitrogen for transportation and then frozen at -70 C until measurements of tissue malondialdehyde (MDA) and glutathione (GSH) levels. The remaining halves of the left kidneys and right kidneys were fixed in 10% formalin. The changes which developed during the ischemia-reperfusion period in the left kidney were investigated by histopathological examination and compared with those of the normal contralateral kidney. When compared with the control group, tissue MDA and GSH levels were similar in the midazolam group (p > 0.05). Tubular damage with tubulitis and focal interstitial inflammatory infiltration were observed in histopathological examinations of reperfused left kidneys of the control group. There was PMNL infiltration only in perirenal fat tissue of the midazolam group. Right kidneys were histopathologically normal in both groups. We concluded that within this dosage midazolam does not have any antioxidant effect in reperfused rat kidneys following ischemia. PMID- 12549655 TI - Review of augmentation 'clam' cystoplasy in a district general hospital setting. AB - Eighteen patients underwent augmentation 'clam' cystoplasty over a three-year period. All patients had urodynamic proven detrusor instability, which was refractory to conservative management. Fifteen of the patients are dry and appliance free. We performed more operations than have been reported in previous studies from other hospitals serving a similar population. Clam cystoplasty is an operation with an excellent outcome, which should be offered to patients with detrusor instability refractory to conservative management. PMID- 12549657 TI - Massive and disproportionate elevation of blood urea nitrogen in acute azotemia. AB - In renal failure, blood urea nitrogen and serum creatinine usually rise in tandem; the normal BUN: Cr ratio is 10-15: 1. Disproportionate rises in BUN: Cr (> 20: 1) often imply pre-renal azotemia but may be caused by increased protein catabolism or an excessive protein load. In this study we looked at intensive care patients who acutely developed markedly increased BUN (> or = 100 mg/dL) with only modest elevation of Cr (< or = 5 mg/dL) for possible causes of the disproportionate azotemia. There were 19 such cases collected over 6 months, nine women and ten men, with mean age 69.2 +/- 4.4 years (13/19 > 75 years). Peak BUN was 156 +/- 11 mg/dL; peak Cr 4.3 +/- 0.5 mg/dL. Eleven patients expired. Mean serum albumin at the time of consultation was 2.7 +/- 0.2 g/dL; mean total lymphocyte count 1.0 +/- 0.1/mm3. Of possible factors causing the azotemia, nine patients had documented hypovolemia; eight had congestive heart failure; six were in septic or hypovolemic shock, and two received high-dose steroids. As contributing factors, eight patients had Salb < 2.5 g/dL; eight were given a high protein intake > 100 g/d; two had HIV, and two others had gastrointestinal bleeding. Infection was present in 14 patients; seven had sepsis (bacteremia with hypotension). All patients had at least one of these factors present and 16/19 had two or more. Fractional Na excretion was < 1% (consistent with pre-renal azotemia) in only four of the 11 patients in whom it was measured. We conclude that severely disproportionate BUN : Cr is frequently multifactorial and is most common in the elderly, perhaps due to their lower muscle mass, and in ICU patients given a high protein intake. It is often not indicative of uncomplicated renal hypoperfusion, although low renal perfusion (hypovolemia, shock, or heart failure) is common. Mortality is high due to the severe illnesses, especially infection, worsened by decreased renal function and hypercatabolic state. PMID- 12549658 TI - Elderly patients on chronic hemodialysis: effect of the secondary hyperparathyroidism on the hemoglobin level. AB - In patients on chronic hemodialysis (CHD) hyperparathyroidism (HPTH) is associated with anemia and resistance to erythropoietin (EPO). This study included 86 CHD elderly pts (mean age 74.8 y, mean time on CHD = 50.5 mos); they were divided into two groups: I (n = 31) - PTH > 250 pg/mL and II (n = 55) - PTH < 250 pg/mL. All these patients had been on CHD for > 6 mos. No differences were found between groups in respect to age, sex distribution and time on CHD. The levels of creatinine, BUN, Ca, Al, Fe, albumin and ferritin were similar. Group I had a higher P level (5.4 vs 4.3 mg/dL, p = 0.001) and Ca x P (53.5 vs 43.7, p = 0.009). Also the Hct (31 vs 33.5%, p = 0.008) and the Hb (10.4 vs 11.2 g/dL, p = 0.009) values were lower in Group I. The EPO dose (88 vs 85 U/kg/week, p = ns) was similar in the two groups. Our data showed that elderly patients with HPTH have lower Hct and Hb levels than do younger patients on a similar EPO dose. We believe these patients will need a more aggressive therapy with calcitriol. PMID- 12549656 TI - Renal stone disease, elevated iPTH level and normocalcemia. AB - BACKGROUND: There is a well established relationship between primary hyperparathyroidism and recurrent calcium-containing calculi. Traditionally, the diagnosis is confirmed by the presence of elevated intact parathyroid hormone (iPTH) and serum ionised calcium levels. The prevalence and role of elevated iPTH in the presence of normocalcemia in patients with renal stone disease is poorly understood. The aim of the present study was to describe the findings in patients who had renal stone disease, an elevated iPTH level and normocalcemia. METHODS: During the last decade, 414 patients, who had normal renal function, were investigated and treated for renal calculi in the Renal Stone Clinic of the Toronto Western Hospital. Of these 414 patients, 40 (9.6%) had an elevated intact iPTH level and normal serum calcium (total and ionised) on repeated measurements. In all these patients we performed detailed clinical and laboratory investigations to determine risk factors for stone formation. Correlation analysis was done using Pearson test and the weights of factors influencing iPTH level were compared using multiple regression analysis. RESULTS: The average duration of a history of stone disease was 12.0 +/- 10.5 years. Most of these patients had passed their stones spontaneously, 15 underwent lithotripsy, in six the stones were removed by basket catheters and one patient each had partial nephrectomy, nepholithotomy or uretero-lithotomy. Twelve had a positive family history, two had history of intestinal malabsorption and one patients had a history of immobilisation. All of these patients had elevated serum parathyroid hormone in the range of 3% to 134% (median 20.5%) above upper limit of normal (F = M); all had normal serum total and ionised calcium and normal urine excretion of calcium (except in one). Additional risk factor for stone formation included: low level of 25-hydroxyvitamin D in four patients, low output and high urine osmolality in four patients, high urine sodium in nine and high oxalate excretion in eight patients. Citrate excretion was low in seven, magnesium excretion in six patients and tubular reabsorption of phosphate in 22 patients. Urine hydroxyprolin was increased in two and decreased in four patients. Combined abnormalities were found in 14 while 17 patients did not have any abnormality apart from elevated iPTH level. Multiple regression analysis did not suggest that any of the selected predictors had a significant influence on PTH release. CONCLUSIONS: 9.6% of patients with recurrent kidney stones had normocalcemia and elevated iPTH level in the presence of normal renal function. The study did not show any distinct pattern of abnormalities that would suggest a mechanism of stone disease in these patients. Further investigations are necessary to determine the significance of elevated iPTH in normocalcemic patients with renal stone disease and establish whether we should consider neck exploration for parathyroidectomy in these patients. PMID- 12549659 TI - Vasculitides with predominantly renal involvement: influence of age on the mode of presentation. AB - Vasculitis with predominant renal involvement are increasingly observed in elderly patients. It predominantly involves patients with microscopic polyangiitis, Wegener's granulomatosis, renal limited vasculitis and, in a smaller proportion, Churg Strauss syndrome. The elderly patients present very few obvious symptoms or signs of vasculitis such as hemoptysis and pulmonary infiltrates, and they are often submitted to: 1/renal biopsy when renal failure is already advanced; 2/ have more severe renal disease than do young and middle aged patients (81% of acute and/or rapidly progressive renal failure versus 62%, and 30% of oliguria versus 6%); and 3/ a worse prognosis compared with younger individuals (probability of survival at 3 months, 1 year and 5 years of 93%, 93% and 92% in patients under 65, versus 67%, 62% and 46% in the elderly). Management of all patients with vasculitis and predominant renal involvement requires a quick diagnosis and immunosuppressive treatment aimed at controlling the disease. Treatment is mantained for 18 to 24 months. Nevertheless, relapses do occur (between 20% to 52%) of all patients with microscopic polyangiitis and Wegener's granulomatosis. PMID- 12549660 TI - Factors influencing treatment decisions of hazardous waste generators: a study of factories in the Bangkok region. AB - While the developed countries are engaged in developing new technology for waste avoidance, recycling and reuse, developing countries are still wrestling to decide on the best option to treat and dispose of wastes. This is particularly so in the case of hazardous industrial wastes (HIW). In order to ensure safe HIW management, one policy response has been to make available a central treatment facility for HIW generators. Such a facility was established in Thailand--the Bang Khuntien Treatment Facility (BKTF)--for the HIW generators in the Bangkok region. However, this facility operates much below its capacity. A study was therefore undertaken between April 1996 and April 1997 to investigate the factors that influence the HIW producing factories' decisions regarding the use of this facility. The survey reveals a distinct pattern: the majority of the non-users of the facility are small- and medium-scale factories, small generators of hazardous wastes, non recipients of Board of Investment (BOI) concessions, and located far away from the facility. The results from a multiple regression analysis show that a factory decision on usage of the facility is positively influenced by its size, amount of wastes generated, knowledge of the facility, pressure from regulators, and consideration of employee and community welfare. It is negatively influenced by distance and costs of using the treatment facility. PMID- 12549661 TI - Kinetics of CH4 oxidation in mixed culture. AB - Microbial cultures used for methane oxidation experiments were prepared by enriching under a 20 % CH4 atmosphere either leachate extracts from two different landfill sites or a digested sewage sludge. No microbes were axenically isolated, therefore the cultures were mixtures of CH4 oxidising and other aerobic microorganisms. CH4 oxidation experiments were carried out with a batch reactor, whose change in gas volume could be measured. A mixture of air and methane with CH4 concentrations ranging from 5 to 20 % was introduced into the reactor. Methane oxidation rate was expressed as a Monod equation for CH4 and a first order reaction for O2. Maximum oxidation rate (Vmax) resulted 2.1 x 10(-13) to 3.4 x 10(-11) hr(-1) cell(-1), and half saturation constant (KM) was 5.1 x 10(-5) to 6.5 x 10(-4) mol L(-1). The molar ratio of consumed O2 to oxidised CH4 was 1.5 1.8 PMID- 12549662 TI - Evaluation of the potentialities to reduce greenhouse gases (GHG) emissions resulting from various treatments of municipal solid wastes (MSW) in moist tropical climates: application to Yaounde. AB - The authors here analyse the emission of greenhouse gases (GHG) resulting from the various treatment of municipal solid waste found in the town of Yaounde. Four management systems have been taken as the basis for analyses. System 1 is the traditional collection and landfill disposal, while in system 2 the hiogas produced in the landfill is recuperated to produce electricity. In systems 3 and 4, in addition to the collection, we have introduced a centralised composting or biogas plant before the landfilling disposal of refuse. A Life Cycle Inventory (LCI) of the four systems was made; this enable us to quantify the flux of matter and of energy, consumed or produced by the systems. Following this, only the greenhouse effect was taken into account to evaluate the ecological consequences of the MSW management systems. The method used to evaluate this impact takes into consideration on the one hand, GHG emissions or avoided emission following the substitution of fuel with methane recovered from landfills or produced in the digesters, and on the other hand, sequestrated carbon in the soil following the regular deposit of compost. Landfilling without recuperation of methane is the most emitting solution for greenhouse gas: it leads to the emission of 1.7 ton of carbon dioxide equivalent (tCO2E) per ton of household waste. Composting and methanisation allow one to have a comparable level of emission reduction, either respectively 1.8 and 2 tCO2E/t of MSW. In order to reduce the emission of GHG in the waste management systems, it is advisable to avoid first of all the emissions of methane coming from the landfills. System 2 seems to be a solution that would reduce the emissions of GHG at low cost (2.2 to 4 $/tCO2E). System 2 is calculated as the most effective at the environmental and economic level in the context of Yaounde. Therefore traditional collection, landfill disposal and biogas recuperation to produce electricity is preferable in moist tropical climates. PMID- 12549663 TI - Modelling for environmental assessment of municipal solid waste landfills (part II: biodegradation). AB - The biodegradation module of a simulation program for municipal solid waste landfills (MODUELO) was developed. The biodegradation module carries out the balance of organic material starting with the results of the hydrologic simulation and the waste composition. It simulates the biologic reactions of hydrolysis of solids and the gasification of the dissolved biodegradable material. The results of this module are: organic matter (COD, BOD and elemental components such as carbon, hydrogen, nitrogen, oxygen, sulfur and ash), ammonium nitrogen generated with the gas and transported by the leachates and the potential rates of methane and carbon dioxide generation. The model was calibrated by using the general tendency curves of the pollutants recorded in municipal solid waste landfills, fitting the first part of them to available landfill data. Although the results show some agreement, further work is being done to make MODUELO a useful tool for real landfill simulation. PMID- 12549664 TI - Molecular characterisation of bacteria in a wetland used to remove ammoniacal-N from landfill leachate. AB - Bacteria associated with leachate from a constructed, surface-flow wetland effective at removing ammonium-N, were characterised using molecular methods which bypass the need for cultivation. DNA was extracted from the sample and bacterial 16S rDNA sequences amplified and cloned. Ninety-six individual clones were re-amplified and analysed by restriction digestion and targeted sequencing. A collector's curve was used to estimate the total number of bacterial groups (operational taxonomic units, OTUs) in the sample at 47. Thirty-six clones representing 28 OTUs were sequenced and characterised by aligning them against published sequences. Of these, 10 were confidently identified, whilst for the remaining 18, the closest match was obtained. The sequences in 64% of the OTUs were similar to those of the Proteobacteria but no classical nitrifying bacteria were identified. However, alkaliphilic Methylomicrobium sp. that oxidise ammonium and bacteria from the genus Alcaligenes, at least one species of which is capable of heterotrophic nitrification, were detected. PMID- 12549665 TI - Carbide sludge management in acetylene producing plants by using vacuum filtration. AB - Carbide sludge (10.4-11.5 tonnes day(-1)) is generated from the reaction of calcium carbide (900 kg) and water (6,000 L) in the production of acetylene (2,400 m3), in three selected acetylene manufacturing plants. The sludge (of pH 12.2 and containing Cu, Pb, Fe, Mn, Ni and Zn ions whose concentrations exceed the Department of Environment limits for industrial wastewater) was treated by vacuum filtration as a substitute for the ponding system, which is environmentally less acceptable. A similar system by flocculation was also developed. The filtration system represents an improvement over the ponding method, as shown by a pH of 7 for the clear filtrate; the solid cake, which contains 98% of the metals, can be conveniently disposed at an integrated scheduled waste treatment centre. PMID- 12549666 TI - Toxicity assessment of a dye industry treatment sludge. AB - Industrial treatment sludges often contain heavy metals and a variety of other hazardous substances which can cause soil and underground water pollution. In the present work, sludge samples from a dye industry treatment plant were analysed for their heavy metal contents, and three different leaching procedures were used to assess the potential toxicity of the sludge. The sludge samples were also incinerated at 600 degrees C, and the remaining inorganic residues were also tested for their leaching characteristics by using the same leaching procedures. The treatment sludge investigated in this study contained about 55 grams of Fe per kilogram of dry sludge because iron salts are used in the chemical treatment of the wastewater. Excluding iron, the decreasing order of abundance for heavy metals in the sludge samples was found to be: Zn> Mn> Cr> Cu> Ni> Pb> Co> Cd. The average concentration of each of these heavy metals varied between 25-740 mg/kg of dry sludge. The Fe content of the inorganic residues was found to be about 140 g/kg, and the heavy metal content varied between 40-2,800 mg/kg. In all the leaching tests, Pb was the most readily extracted metal from the sludge, whereas from the inorganic residue, Cd was leached the most easily. The toxicity of the sludge is estimated according to pertinent legislation. PMID- 12549667 TI - Fluidised bed catalytic pyrolysis of scrap tyres: influence of catalyst:tyre ratio and catalyst temperature. AB - Pyrolysis with on-line Zeolite catalysis of scrap tyres was undertaken in a fluidised bed reactor with the aim of maximising the production of higher value single ring aromatic hydrocarbons in the derived oil. Experiments were carried out in relation to the ratio of the catalyst to tyre feedstock and the temperature of the catalyst bed. Two Zeolite catalysts were examined, a Y-type Zeolite catalyst and Zeolite ZSM-5 catalyst of differing pore size and surface activity. The composition of the oils derived from the uncatalysed fluidised bed pyrolysis of tyres showed that benzene concentration was 0.2 wt%, toluene concentration was 0.8 wt%, o-xylene was 0.3 wt%, m/p-xylenes were 1.8 wt% and limonene was 4.3 wt%. Benzene, toluene and xylenes present in the oils showed a significant increase in the presence of both of the catalysts. The maximum concentrations of these chemicals for the Y-Zeolite (CBV-400) catalyst was 1 wt% for benzene, 8wt% for toluene, 3 wt% for o-xylene and 8.5 wt% for m/p-xylenes, produced at a catalyst:tyre ratio of 1.5. There was less influence of catalyst temperature on the yield of benzene, toluene and xylenes, however, increasing the temperature of the catalyst resulted in a marked decrease in limonene concentration. The Y-type Zeolite catalyst produced significantly higher concentrations of benzene, toluene and xylenes which was attributed to the larger pore size and higher surface acidity of the Y-Zeolite catalyst compared to the Zeolite ZSM-5 catalyst. PMID- 12549668 TI - Review: mercury in waste incineration. AB - The paper investigates the sources of mercury (Hg) in municipal/industrial waste and the consequences of the presence of this pollutant for the incineration of this waste. About 1990 the average mercury concentration of the feed stream to incinerators was about 4 mg kg(-1). The concentration decreased considerably during the last decade thanks to a considerable reduction of the application of mercury and to the introduction of effective battery return systems. Presently the mercury concentration in municipal SOLID waste is approximately 2 mg kg(-1). During incineration mercury passes practically for 100% in the flue gas. The techniques for mercury removal from flue gases are discussed at the hand of practical examples. It is concluded that there are a number of processes which guarantee mercury concentrations of <50 microg Nm(-3) in the clean gas, the present emission limit concentration. All mercury control processes produce a new solid or liquid waste stream that contains the mercury removed from the flue gas. This stream has to be disposed of as hazardous waste in a qualified landfill. The flue gas from waste incinerators undergoes very rapid dispersion and dilution after leaving the incinerator stack. It follows that the maximum mercury concentration in the ambient air will remain at least five to six orders of magnitude below the lowest MAC value (=Maximum Admissible Concentration in work spaces) and that public health will not be threatened. PMID- 12549669 TI - Recent advances in molecular diversity. PMID- 12549670 TI - An overview of the diversity represented in commercially-available databases. PMID- 12549671 TI - Filtering databases and chemical libraries. PMID- 12549672 TI - Current trends in lead discovery: are we looking for the appropriate properties? AB - The new drug discovery paradigm is based on high-throughput technologies, both with respect to synthesis and screening. The progression HTS hits --> lead series --> candidate drug --> marketed drug appears to indicate that the probability of reaching launched status is one in a million. This has shifted the focus from good quality candidate drugs to good quality leads. We examined the current trends in lead discovery by comparing MW (molecular weight), LogP (octanol/water partition coefficient, estimated by Kowwin) and LogSw (intrinsic water solubility, estimated by Wskowwin) for the following categories: 62 leads and 75 drugs; compounds in the development phase (I, II, III and launched), as indexed in MDDR; and compounds indexed in medicinal chemistry journals, categorized according to their biological activity. Comparing the distribution of the above properties, the 62 lead structures show the lowest median with respect to MW (smaller) and LogP (less hydrophobic), and the highest median with respect to LogSw (more soluble). By contrast, over 50% of the medicinal chemistry compounds with activities above 1 nanomolar have MW > 425, LogP > 4.25 and LogSw < -4.75, indicating that the reported active compounds are larger, more hydrophobic and lesssoluble when compared to time-tested quality leads. In the MDDR set, a progressive constraint to reduce MW and LogP, and to increase LogSw, can be observed when examining trends in the developmental sequence: phase I, II, III and launched drugs. These trends indicate that other properties besides binding affinity, e.g., solubility and hydrophobicity, need to be considered when choosing the appropriate leads. PMID- 12549673 TI - Multiobjective optimization of combinatorial libraries. AB - Combinatorial chemistry and high-throughput screening have caused a fundamental shift in the way chemists contemplate experiments. Designing a combinatorial library is a controversial art that involves a heterogeneous mix of chemistry, mathematics, economics, experience, and intuition. Although there seems to be little agreement as to what constitutes an ideal library, one thing is certain: only one property or measure seldom defines the quality of the design. In most real-world applications, a good experiment requires the simultaneous optimization of several, often conflicting, design objectives, some of which may be vague and uncertain. In this paper, we discuss a class of algorithms for subset selection rooted in the principles of multiobjective optimization. Our approach is to employ an objective function that encodes all of the desired selection criteria, and then use a simulated annealing or evolutionary approach to identify the optimal (or a nearly optimal) subset from among the vast number of possibilities. Many design criteria can be accommodated, including diversity, similarity to known actives, predicted activity and/or selectivity determined by quantitative structure-activity relationship (QSAR) models or receptor binding models, enforcement of certain property distributions, reagent cost and availability, and many others. The method is robust, convergent, and extensible, offers the user full control over the relative significance of the various objectives in the final design, and permits the simultaneous selection of compounds from multiple libraries in full- or sparse-array format. PMID- 12549674 TI - Predictive QSAR modeling based on diversity sampling of experimental datasets for the training and test set selection. AB - One of the most important characteristics of Quantitative Structure Activity Relashionships (QSAR) models is their predictive power. The latter can be defined as the ability of a model to predict accurately the target property (e.g., biological activity) of compounds that were not used for model development. We suggest that this goal can be achieved by rational division of an experimental SAR dataset into the training and test set, which are used for model development and validation, respectively. Given that all compounds are represented by points in multidimensional descriptor space, we argue that training and test sets must satisfy the following criteria: (i) Representative points of the test set must be close to those of the training set; (ii) Representative points of the training set must be close to representative points of the test set; (iii) Training set must be diverse. For quantitative description of these criteria, we use molecular dataset diversity indices introduced recently (Golbraikh, A., J. Chem. Inf. Comput. Sci., 40 (2000) 414-425). For rational division of a dataset into the training and test sets, we use three closely related sphere-exclusion algorithms. Using several experimental datasets, we demonstrate that QSAR models built and validated with our approach have statistically better predictive power than models generated with either random or activity ranking based selection of the training and test sets. We suggest that rational approaches to the selection of training and test sets based on diversity principles should be used routinely in all QSAR modeling research. PMID- 12549675 TI - Reactant- and product-based approaches to the design of combinatorial libraries. PMID- 12549677 TI - New methods in predictive metabolism. PMID- 12549676 TI - Towards a new age of virtual ADME/TOX and multidimensional drug discovery. AB - With the continual pressure to ensure follow-up molecules to billion dollar blockbuster drugs, there is a hurdle in profitability and growth for pharmaceutical companies in the next decades. With each success and failure we increasingly appreciate that a key to the success of synthesized molecules through the research and development process is the possession of drug-like properties. These properties include an adequate bioactivity as well as adequate solubility, an ability to cross critical membranes (intestinal and sometimes blood-brain barrier), reasonable metabolic stability and of course safety in humans. Dependent on the therapeutic area being investigated it might also be desirable to avoid certain enzymes or transporters to circumvent potential drug drug interactions. It may also be important to limit the induction of these same proteins that can result in further toxicities. We have clearly moved the assessment of in vitro absorption, distribution, metabolism, excretion and toxicity (ADME/TOX) parameters much earlier in the discovery organization than a decade ago with the inclusion of higher throughput systems. We are also now faced with huge amounts of ADME/TOX data for each molecule that need interpretation and also provide a valuable resource for generating predictive computational models for future drug discovery. The present review aims to show what tools exist today for visualizing and modeling ADME/TOX data, what tools need to be developed, and how both the present and future tools are valuable for virtual filtering using ADME/TOX and bioactivity properties in parallel as a viable addition to present practices. PMID- 12549679 TI - Chemoinformatics methods for systematic comparison of molecules from natural and synthetic sources and design of hybrid libraries. AB - Until recently, the field of diversity and library design has more or less ignored natural products as a compound source. This is probably due to at least two reasons. First, combinatorial and reaction-based approaches have been major focal points in the early days of computational library design. In addition, a widespread view is that natural products are often highly complex and not amenable to medicinal chemistry efforts. This contribution introduces recent computational approaches to systematically analyze natural molecules and bridge the gap between natural products and synthetic chemistry programs. Large scale comparisons of natural and synthetic molecules are discussed as well as studies designed to identify 'synthetic mimics' of natural products with specific activity. In addition, a concept for the design of natural/synthetic hybrid libraries is introduced. Although research in this area is still in its early stages, an important lesson to be learned from computational analyses is that there is no need to a priori 'shy away' from natural products as a source for molecular design. PMID- 12549680 TI - Value of keeping records of mortality. AB - OBJECTIVE: To evaluate treatment and complications which is essential for good medical practice. DESIGN: Prospective audit. SETTING: City hospital, The Netherlands. SUBJECTS: All the patients who died on the surgical ward between 1994 and 1998 and were classified according to four categories of mortality recording. INTERVENTIONS: The causes of death, inaccuracies in treatment and the extent of agreement between premortem and postmortem findings were documented. MAIN OUTCOME MEASURES: Morbidity and mortality. RESULTS: Of the 11,195 patients admitted, 420 (4%) deceased during their hospital stay. Most patients died of the disease with which they presented at admission (n = 176, 42%) or of complications (n = 167, 40%). In 20% (n = 83) of the cases a shortcoming in the clinical course was found. 251 of the 420 patients who died (60%) had a necropsy. 53 of the 251 reports (21%) gave information that could have had an effect on the treatment or the clinical course. CONCLUSIONS: Recording mortality is a way of testing the diagnostic and therapeutic accuracy in our quest for a high quality of care. PMID- 12549678 TI - Exploring privileged structures: the combinatorial synthesis of cyclic peptides. AB - Head-to-tail cyclic peptides have been reported to bind to multiple, unrelated classes of receptor with high affinity. They may therefore be considered to be privileged structures. This review outlines the strategies by which both macrocyclic cyclic peptides and cyclic dipeptides or diketopiperazines have been synthesised in combinatorial libraries. It also briefly outlines some of the biological applications of these molecules, thereby justifying their inclusion as privileged structures. PMID- 12549681 TI - Repeat antireflux surgery: effectiveness of a toupet partial posterior fundoplication. AB - OBJECTIVE: To report the feasibility and efficacy of posterior partial fundoplication for patients with recurrent reflux after previous antireflux operations. DESIGN: Prospective non-randomised single hospital study. SETTING: University hospital, Sweden. PATIENTS: 32 consecutive patients who required reoperation after failure of previous operations for gastro-oesophageal reflux disease. MAIN OUTCOME MEASURES: Control of symptoms, oesophageal acid reflux. RESULTS: Most patients were referred for reoperation because of recurrent reflux, but six patients had a symptomatic intrahoracic herniation of the wrap with a life-threatening perforation in one patient. Three patients had recurrent reflux within 6 months of the operation and another two were classified as failures. The remaining 27 (84%) were classified as having a successful outcome during a mean follow-up period of 20 months. CONCLUSION: A posterior partial fundoplication can be done safely in a repeat operation for patients whose antireflux operation has failed. PMID- 12549682 TI - Factors that predict morbidity and mortality in patients with perforated peptic ulcers. AB - BACKGROUND: The demographic changes of perforated peptic ulcer disease were assessed in a well-defined population in northern Finland. The high mortality from perforated peptic ulcer underlines the importance of risk stratification, and clinical variables and three scoring systems were therefore tested for their ability to predict the probability of morbidity and mortality. METHODS: Two hundred and eighty patients admitted to a university hospital with peptic ulcer perforation during the 22-year period 1979-2000 were identified using a computer database, and their clinical data were reviewed from the database and patient records. RESULTS: The annual operation rate for ulcer perforation has varied from 2.7/100,000 in 1979 to 6.2 in 1985 and 3.6 in 2000 without any significant changes during the examination period. More operations have been performed for duodenal (157) than for gastric (123) ulcer perforations, but their incidence rates were quite similar at the beginning and the end of the period. Of the clinical variables, two or more associated illnesses, duration of symptoms for more than 24 hours, the amount of abdominal liquid, and low albumin concentration predicted morbidity, while a long duration of symptoms and the amount of abdominal liquid were independent risk factors for mortality. The MPI score correctly predicted 96% of postoperative complications and all the three scores, i.e. the Boey score, the MPI score and the ASA score, were good predictors of mortality. CONCLUSION: The incidence of peptic ulcer perforation has not changed during the 22-year period in northern Finland. All the three scores, i.e. the Boey score, the MPI score and the ASA score, predict mortality in patients with peptic ulcer perforation and are suitable for risk stratification preoperatively in the surgical ward. PMID- 12549683 TI - Lichtenstein repair for incarcerated groin hernias. AB - OBJECTIVE: To evaluate the use of Lichtenstein tension-free repair for incarcerated groin hernias. DESIGN: Prospective consecutive series. SETTING: Tertiary academic surgery department, Poland. SUBJECTS: 27 patients whose incarcerated groin hernias were operated on between November 1997 and June 2001. INTERVENTION: Tension-free hernia repair by the Lichtenstein technique. MAIN OUTCOME MEASURES: Morbidity, recurrence rates and mortality. RESULTS: There were 2 postoperative deaths unrelated to implantation of the mesh (myocardial and cerebral infarction). In 1 patient a subcutaneous fluid collection was noted and was successfully treated by evacuation of the fluid. The remaining 24 patients had no complications during follow-up (mean 18 months, range 2-42 months). CONCLUSION: Monofilament polypropylene mesh can be successfully used not only in elective operations, but also in emergency operations for incarcerated inguinal hernias. PMID- 12549684 TI - Comparison of local, spinal, and general anaesthesia for inguinal herniorrhaphy. AB - OBJECTIVE: To compare local, spinal, and general anaesthesia for inguinal hemiorraphy in otherwise healthy patients with respect to duration of operation, time in operating room, postoperative pain, complications, rehabilitation, and satisfaction. DESIGN: Prospective randomised controlled trial. SETTING: University hospital, Turkey. SUBJECTS: Seventy-five men with unilateral primary inguinal hernias, Nyhus type II and III, and ASA I and II. INTERVENTIONS: Lichtenstein repairs with standard local, spinal, or general anaesthesia. MAIN OUTCOME MEASURES: Duration of operation and anaesthesia, postoperative pain scores, analgesic requirements, complications, length of hospital stay, postoperative rehabilitation, and satisfaction. RESULTS: With local anaesthesia, we recorded shorter time spent in the operating room, lower incidence of nausea and urinary retention, and more satisfaction. In the local and spinal anaesthetic groups, postoperative analgesic requirements and length of hospital stay were less than in the general anaesthesia group. CONCLUSIONS: Local anaesthesia is suitable for day-case hernia repair with fewer postoperative problems and less analgesia requirement. Patients also reported greater satisfaction. Local anaesthesia may be preferred to other methods. PMID- 12549685 TI - Hernias of the abdominal wall in patients over the age of 70 years. AB - OBJECTIVE: To find out if the patients' age affects the treatment of abdominal hernias and the results in relation of the age increase. DESIGN: Retrospective and prospective study. SETTING: University hospital, Spain. SUBJECTS: 664 patients aged 70 years or more operated on for abdominal hernia between 1986 1998. Patients were divided into three groups: 443 aged 70-79; 202 aged 80-89; and 19 patients aged 90 years or more. MAIN OUTCOME MEASURES: Perioperative risk, type of surgery and deaths. RESULTS: 117 women (52%) had femoral hernias, compared with 32 men (7%) (p = 0.0001). The incidence of femoral hernia over 80 years of age was 79/221 (36%) compared with 70/443 (16%) among patients in their seventies (p = 0.0001). 97 of the patients aged 70-79 (22%) were operated on as emergencies, 107 of those aged 80-89 (53%), and 17 in patients 90 or older (89%, p = 0.0001). The mortality rate was 1% in the 70-79 group (n = 6), 5% (n = 10) in the 80-89 group, and 3/19 died in the over 90 group (p = 0.0001). No deaths were reported after elective surgery. CONCLUSION: Emergency operations in elderly patients with abdominal wall hernias are increasingly more common as the patient get older. As result, there is an unacceptable increase in postoperative mortality. PMID- 12549686 TI - Influence of laparoscopic cholecystectomy on the prevalence of operations for gallstones in Norway. AB - OBJECTIVE: To assess trends in the treatment of gallstone disease, by recording the number of admissions to hospital and operations; to find out if there are regional differences in the number of operations for gallstone disease; to compare these with those of other countries, and to discuss possible causative factors with special reference to introduction of laparocopic cholecystectomy. DESIGN: Study of medical records. SETTING: University hospital, Norway. SUBJECTS: A total of 55,622 hospital stays and 28,048 operations for gallstone disease were recorded in a national registry for the 8-year period 1992-9, which comprise 99% of all admissions for gallstone disease. INTERVENTIONS: Data were compared for number of admissions to hospitals for gallstone disease, incidence of different operations, and trends in treatment. Different regions in Norway were also compared. RESULTS: Both the number of hospital stays and the number of gallstone operations increased slightly over the period. Compared with 1992, the open cholecystectomy rate was reduced to 25%, and laparoscopic operations increased more than four times. The incidence of endoscopic procedures showed an annual increase from 1992-9, followed by a fall in 1999 to the 1992 level. CONCLUSION: Treatment trends show a pronounced increase in laparoscopic operations, and at the same time fewer open operations. The total number of operations has increased slightly in Norway. The prevalence of operations for gallstones in Norway is the lowest reported in Western Europe. The overall number of gallstone operations in Norway is relatively small. The increase in the number of operations for gallstones in Norway was mainly caused by the increase in laparoscopic cholecystectomies. After a steady rise, there was a fall in the number of endoscopic procedures for common bile duct stones during the last year. At the same time, however, magnetic resonance cholangiopancreatography was introduced in Norway. The prevalence of gallstone disease in the population studied seems to have had little influence on the incidence of operations. Many Norwegian patients with gallstone disease who could benefit from operation are probably not being offered surgical treatment today, but are treated in an expectant manner instead. PMID- 12549687 TI - Teaching laparoscopic cholecystectomy: do beginners adversely affect the outcome of the operation? AB - OBJECTIVE: To evaluate the safety of elective laparoscopic cholecystectomy (LC) as a supervised teaching procedure. DESIGN: Prospectively documented series, retrospective analysis. SETTING: University teaching hospital, Austria. SUBJECTS: 581 consecutive patients who underwent elective LC for symptomatic cholelithiasis between January 1993 and December 1997. INTERVENTIONS: LC were allocated to three groups: the first (n = 91) were done by supervised beginners (who had done fewer than 11 LC), the second (n = 249) by supervised trainees who had a little experience (they had done more than 10 but fewer than 51 LC), and the third group (n = 241) who were experienced surgeons (they had done more than 50 LC). MAIN OUTCOME MEASURES: Minor and major complications, conversion and reoperation rate, length of operation and postoperative hospital stay. RESULTS: The minor intraoperative complication rates were 36/91 (40%), 115/249 (46%) and 49/241 (20%), respectively (p < 0.001 when experienced surgeons were compared with the 2 trainees' groups). There were no significant differences between the three groups regarding major complications (1/91, 4/249 and 4/241), conversions (5/91, 21/249, and 17/241) and reoperation rate (1/91, 3/249 and 3/241), median (range) length of operation 82 (24-159), 84 (25-249) and 82 (21-234) minutes and hospital stay 4 (3-19), 4 (3-11) and 4 (2-15) days. CONCLUSION: Elective LC for symptomatic cholelithiasis done by trainees under supervision does not increase surgical morbidity. PMID- 12549688 TI - Early operation or conservative management of patients with small bowel obstruction? AB - OBJECTIVE: To evaluate the outcome after initial non-operative treatment in patients with small bowel obstruction (SBO). DESIGN: Prospective study. SETTING: University hospital, Norway. PATIENTS: One hundred and fifty-four patients with 166 episodes of SBO admitted during the period (1994-1995). Patients younger than 10 years as well as patients with large bowel obstruction, paralytic ileus, incarcerated hernia or SBO caused by cancer were excluded from the study. INTERVENTIONS: Patients with signs of strangulation were operated on early. The rest were given a trial of conservative treatment. MAIN OUTCOME MEASURES: Need of operative treatment. Incidence of bowel strangulation, complications and death. RESULTS: There were 166 cases of SBO. Twenty patients were operated on early among whom bowel was strangulated in 9. Among the 146 patients initially treated conservatively 93 (64%) settled without operation, 9 (6%) had strangulated bowel and 3 (2%) died. Of the 91 patients with partial obstruction but no sign of strangulation, 72 (79%) resolved on conservative treatment. CONCLUSIONS: Patients with partial obstruction with no sign of strangulation should initially be treated conservatively. When complete obstruction is present, it may settle on conservative management, but the use of supplementary diagnostic tools might be desirable to find the patients who will need early operative treatment. PMID- 12549689 TI - Haemorrhoidectomy: randomised controlled clinical trial of Ligasure compared with Milligan-Morgan operation. AB - OBJECTIVE: To evaluate the efficacy of the Ligasure system in the management of haemorrhoids. DESIGN: Unblinded randomised clinical trial. SETTING: Teaching hospital, Spain. PATIENTS: 112 patients with third and fourth degree haemorrhoids. INTERVENTIONS: For 56 patients we used Ligasure system and a variant of Milligan and Morgan's technique. For the other 56, we used the traditional technique. MAIN OUTCOME MEASURES: Postoperative pain. RESULTS: Operating times varied from 100 seconds for each haemorrhoidal cushion with Ligasure system to the 313 seconds by the traditional technique. The blood loss was not quantifiable in patients operated on with Ligasure. Pain was scored on a visual analogue scale. In the Ligasure group, the mean scores were 4.9 (immediate postoperative period) and 2.3 (24 hours later). In the other group, the scores were 7.8 and 6.9. These differences were significant. CONCLUSION: Haemorrhoidectomy using Ligasure as a technical variant of Milligan and Morgan's technique has important advantages. PMID- 12549690 TI - Long-term results of haemorrhoidectomy. AB - OBJECTIVE: To assess the long-term functional results of Milligan-Morgan haemorrhoidectomy. DESIGN: Retrospective multicentre study. SETTING: One university hospital, one county hospital, and two community hospitals, Sweden. SUBJECTS: 507 of 556 patients who were operated on for haemorrhoids by the Milligan-Morgan technique between January 1987 and December 1995. INTERVENTION: A questionnaire was sent to all 507 patients, the questions in which focused on functional results and satisfaction. MAIN OUTCOME MEASURES: Patients' satisfaction and symptoms of anal incontinence after haemorrhoidectomy. RESULTS: 418 of the 507 responded (82%). Altogether 279 patients (67%) reported a successful result, while 139 patients (33%) reported impaired anal continence. 40 of the 139 patients (29%) claimed that the incontinence was a direct result of the haemorrhoidectomy. Female sex (p = 0.005) and an operation for hygienic problems (p = 0.02) were associated with a higher risk of incontinence. CONCLUSION: Impaired anal continence is common after Milligan-Morgan haemorrhoidectomy and a large proportion of affected patients relate their problems to the operation. PMID- 12549691 TI - Clinical uses of diagnostic peritoneal lavage in stab wounds of the anterior abdomen: a prospective study. AB - OBJECTIVE: To evaluate patients with stab wounds of the anterior abdomen with diagnostic peritoneal lavage (DPL), by using initial aspiration of gross blood from the lavage catheter of more than 10 ml or red blood cell (RBC) count in the lavage fluid of more than 10,000/mm3 as criteria for exploratory laparotomy. DESIGN: Prospective study. SETTING: University hospital, Thailand. PATIENTS: 40 patients who had stab wounds of the anterior abdomen penetrating through the peritoneum but had no obvious indications for immediate exploratory laparotomy. INTERVENTIONS: Diagnostic peritoneal lavage (DPL), exploratory laparotomy. MAIN OUTCOME MEASURES: Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of positive DPL as an indication for exploratory laparotomy. RESULTS: In 27 patients (68%) the DPL was positive and in 13 (33%) it was negative. In 18 patients (67%) the initial aspiration of gross blood was more than 10 ml and in 9 (33%) the RBC count in lavage fluid was more than 10,000/mm3. These 27 patients had exploratory laparotomies, 2 of which were negative. All 13 patients who had negative DPL were observed and discharged home uneventfully. The sensitivity of DPL for positive laparotomy was 100%, the specificity was 87%, the PPV was 93%, the NPV was 100% and the accuracy was 95%. When operative findings of bleeding from the stab wound into the peritoneal cavity were considered as a "negative" laparotomy (n = 7) the sensitivity, the specificity, the PPV, the NPV and the accuracy became 100%, 59%, 67%, 100%, and 78%, respectively. CONCLUSION: The use of DPL in patients with stab wounds of the anterior abdomen, using initial aspiration of gross blood from the lavage catheter of more than 10 ml or RBC count in the lavage fluid of more than 10,000/mm3 as positive criteria for exploratory laparotomy, is safe and practical. PMID- 12549692 TI - Sternal reconstruction with synthetic mesh and metallic plates for high grade tumours of the chest wall. AB - OBJECTIVE: To present our experience of reconstruction of the chest wall after sternectomy for high grade tumours. DESIGN: Retrospective study. SETTING: University hospital, Italy. SUBJECTS: 18 patients who required sternectomy for high grade tumours. INTERVENTIONS: After wide resection of the tumour the sternum and chest wall were reconstructed with polypropylene (Marlex) mesh and mouldable titanium plates. Pedicled muscle flaps were used to complete the reconstruction. MAIN OUTCOME MEASURES: Morbidity, mortality, and outcome. RESULTS: The 18 tumours were chondrosarcomas (n = 7); osteosarcomas, radiation-induced sarcomas, and local recurrences after breast cancer (n = 2 each); and angiosarcoma, Ewing sarcoma, liposarcoma, malignant fibrous histiocytoma, and metastatic renal carcinoma (n = 1 each). One patient died in hospital. Two patients developed wound infections, one required repeated aspirations of a pleural effusion, and one patient later developed loosening of the plate that had been sutured to the clavicular stump. At the time of follow-up (mean 32 months, range 12-74) 16 patients were alive, all of whom had returned to their normal life style. CONCLUSIONS: The technique is easy to follow and has the advantages of a short hospital stay and good local control. It obviates the need for postoperative mechanical ventilation, and means that patients are not prevented from working because of incapacity. PMID- 12549693 TI - Dexamethasone down-regulates endothelial expression of intercellular adhesion molecule and impairs the healing of bowel anastomoses. AB - OBJECTIVE: To find out the role of endothelial expression of intercellular adhesion molecule-1 (ICAM-1) in the healing of intestinal anastomoses in rats, and to establish the effects of peroperative treatment with corticosteroids. DESIGN: Experimental animal study. SETTING: University hospital, Turkey. MATERIAL: 78 Male Wistar rats. INTERVENTIONS: Rats were divided into four groups: Group I, colonic anastomosis only (=18); Group II, colonic anastomosis plus caecal ligation and puncture (=18); Group III, colonic anastomosis plus dexamethasone (=18); and Group IV, colonic anastomosis, plus caecal ligation and puncture, plus dexamethasone (=18). Six animals served as the sham group. The animals underwent bowel transsection and primary anastomosis Infection was produced by caecal ligation and puncture Preoperatively, dexamethasone was given intramuscularly in a dose of 2 mg/kg/day. MAIN OUTCOME MEASURES: After 1, 3 and 5 days, anastomotic healing and endothelial expression of ICAM-1 were measured microscopically. RESULTS: Anastomotic healing was significantly impaired in dexamethasone-treated animals, and endothelial expression of ICAM-1 was reduced. Endothelial expression of ICAM-1 was no higher in the infected group than in controls. Maximum expression of ICAM-1 on endothelial cells was seen on the first day in each group, and declined on the following days, although the sebsequent reduction in expression was not significant. CONCLUSION: Dexamethasone down regulated expression of ICAM-1, which is important in migration of leucocytes from the circulation to the wound site, and significantly impaired the healing of intestinal anastomoses in rats. PMID- 12549695 TI - Giant paratesticular undifferentiated liposarcoma that developed in a long standing inguinal hernia. PMID- 12549694 TI - Neutrophil elastase inhibitor reduces hepatic metastases induced by ischaemia reperfusion in rats. AB - OBJECTIVE: To investigate the possibility in rats that ONO-5046 Na, a new recombinant inhibitor of neutrophil elastase, can reduce hepatic metastases induced by ischaemia-reperfusion. DESIGN: Laboratory experimental study. SETTING: Research laboratory, Japan. SUBJECTS: Male Fischer rats. INTERVENTIONS: Rats underwent 60 min of 70% partial hepatic ischaemia, after which rat colon adenocarcinoma cells (RCN-H4) were injected into the spleen. The animals were divided into two test groups and a control group. One group was given ONO-5046 Na intravenously at 10 mg/kg/hour. A second group was given a saline solution for the same period, while the controls were not made ischaemic. MAIN OUTCOME MEASURES: Three weeks after inoculation, the number of tumour nodules on the liver surface was counted. The anti-cancer effect of ONO-5046 Na was measured by monotetrazolium assay. RESULTS: Hepatic ischaemia-reperfusion increased the number of liver metastases of RCN-H4 in both clamped and unclamped hepatic lobes. ONO-5046 Na significantly inhibited this in unclamped lobes, but had no anti cancer effect. CONCLUSION: Neutrophil elastase may have an important role in increasing haematogenous liver metastases by ischaemia-reperfusion, particularly in unclamped lobes. PMID- 12549696 TI - Pancreatic sarcoidosis mimicking a malignant tumour. PMID- 12549697 TI - Staged treatment of bilobar hepatic metastases from colorectal cancer. PMID- 12549698 TI - Editorial: prevention and intervention science: contributions to developmental theory. PMID- 12549699 TI - Can prevention trials test theories of etiology? AB - This paper addresses the issue of whether prevention research methods, particularly those involving randomized prevention trials, can be used to test theories concerning the etiology of psychopathology. Based on recent empirical and theoretical work in developmental psychopathology, three aspects of etiologic theory are discussed: risk and protective mechanisms, the integration of environmental and genetic factors, and patterns of developmental progression in psychopathology across the life span. It is suggested that integration of prevention trial methods with methods from passive correlational designs, behavioral genetics, and longitudinal studies allows for unique opportunities to test hypotheses about etiology. Empirical literature on the development and prevention of internalizing disorders, particularly depression, is presented to support this argument. Limitations of prevention trials for testing theory are also reviewed. PMID- 12549700 TI - Randomized clinical trials, developmental theory, and antisocial youth: guidelines for research. AB - The design of effective interventions for antisocial adolescents has been greatly influenced by research-based developmental theories. However, randomized clinical trials (RCTs) demonstrating effectiveness of these treatments have contributed minimally to alterations and refinements of these theories. In this article, nine guidelines for research that may enhance the contributions of RCTs to developmental theory are proposed: (1) use mediational analyses to test and refine theories of change; (2) use moderator analyses to test differential pathways to change; (3) assess change processes separately for different types of antisocial behavior; (4) use constructive and dismantling designs to isolate effects of intervention components on targeted mediators and outcomes; (5) assess the time-sensitive nature of change mechanisms with theoretically relevant spacing of multiple assessments and long-term follow-ups; (6) expand the nomological net of developmental theories by assessing intervention effects on nontargeted constructs; (7) use RCTs to disentangle direction of effects questions in developmental theories; (8) conduct secondary analyses of RCTs to assess extra-treatment influences on antisocial behavior; and (9) assess the generalizability of mediator and moderator effects. Use of these guidelines may promote a recursive and iterative relationship between RCTs and theory building. Improved developmental theories may yield more effective interventions, and theory-testing interventions may engender more comprehensive and better informed theories. PMID- 12549701 TI - What violence prevention research can tell us about developmental psychopathology. AB - This article reviews major issues of a developmental psychopathology understanding of youth violence and related insights gained from violence prevention efforts to date. Based in a perspective that emphasizes three areas of knowledge, epidemiology, risk factors and models, and developmental trajectories, findings are reviewed that help clarify theoretical propositions about the developmental patterns, influences, and pathways related to youth violence. Extant research is able to support several major contentions of basic developmental psychopathology theories of youth violence. Support is most robust for risk factors central to prevailing models of the development of youth violence. In addition to noting current knowledge contributions, the review suggests areas of needed additional analytic progress and theoretical extension to refine knowledge and expand understanding of the development of violence risk and its prevention. PMID- 12549702 TI - Interventions as tests of family systems theories: marital and family relationships in children's development and psychopathology. AB - This paper addresses the role of family-based studies of preventive and therapeutic interventions in our understanding of normal development and psychopathology. The emphasis is on interventions designed to improve parent child and/or marital relationships as a way of facilitating development and reducing psychopathology in children and adolescents. Intervention designs provide the gold standard for testing causal hypotheses. We begin by discussing the complexity of validating these hypotheses and the implications of the shift from a traditional emphasis on theories of etiology to developmental psychopathology's newer paradigm describing risks --> pathways --> outcomes. We summarize correlational studies that document the fact that difficult and ineffective parent-child and marital relationships function as risk factors for children's cognitive, social, and emotional problems in childhood and adolescence. We then review prevention studies and therapy evaluation studies that establish some specific parenting and marital variables as causal risk factors with respect to these outcomes. Our discussion focuses on what intervention studies have revealed so far and suggests an agenda for further research. PMID- 12549703 TI - Emotion processes in normal and abnormal development and preventive intervention. AB - We present an analysis of the role of emotions in normal and abnormal development and preventive intervention. The conceptual framework stems from three tenets of differential emotions theory (DET). These principles concern the constructs of emotion utilization; intersystem connections among modular emotion systems, cognition, and action; and the organizational and motivational functions of discrete emotions. Particular emotions and patterns of emotions function differentially in different periods of development and in influencing the cognition and behavior associated with different forms of psychopathology. Established prevention programs have not emphasized the concept of emotion as motivation. It is even more critical that they have generally neglected the idea of modulating emotions, not simply to achieve self-regulation, but also to utilize their inherently adaptive functions as a means of facilitating the development of social competence and preventing psychopathology. The paper includes a brief description of a theory-based prevention program and suggestions for complementary targeted interventions to address specific externalizing and internalizing problems. In the final section, we describe ways in which emotion centered preventions can provide excellent opportunities for research on the development of normal and abnormal behavior. PMID- 12549704 TI - Intervention research, theoretical mechanisms, and causal processes related to externalizing behavior patterns. AB - Intervention research with children and adolescents has suffered from a dearth of relevant theoretical grounding and from the lack of a reciprocal "feedback" mechanism by which clinical trials can inform relevant theorizing and conceptualization. There are hopeful signs, however, of increasing confluence between clinical efforts and theoretical models. Indeed, the key issue I discuss is how intervention studies can yield information about developmental and clinical theory as well as mechanisms related to psychopathology. Specific research examples in the field, particularly those emanating from the Multimodal Treatment Study of Children with attention deficit/hyperactivity disorder (MTA study), reveal that probes of moderator and mediator variables can clearly enhance our knowledge of relevant processes and mechanisms. In fact, recent MTA findings have relevance for models of genetic and epigenetic influence on symptomatology related to attentional deficits and hyperactivity. It would be overzealous, however, to make premature claims regarding etiologic variables from intervention research, as treatment studies typically address variables that are causally far "downstream" from primary causal factors and most clinical trials have statistical power that is barely sufficient for main outcome questions, much less mediational linkages. Overall, the field has severely underutilized experimental intervention research to subserve the dual ends of improving the lives of youth and advancing theoretical conceptualization regarding development and psychopathology. PMID- 12549705 TI - The other side of the coin: using intervention research in child anxiety disorders to inform developmental psychopathology. AB - With advancements in the technology of prevention and treatment of childhood anxiety disorders, information regarding our understanding of normal and abnormal child development can be enriched. Typically, research has focused on developing efficacious and effective interventions with less attention devoted to the impact this information may have on the field of developmental psychopathology. By reviewing the results of both treatment and prevention studies, several potential contributions of intervention research to the field can be explored. Results from wait-list or monitoring control groups will be reviewed, providing valuable information regarding the normal trajectory of the anxious child. Outcomes of children receiving the intervention prove that this pathway can be altered and is not impermeable. Furthermore, a review of long-term follow-up studies addresses the question of whether intervention can change the long-term trajectory of an anxiety-disordered child and prevent disorders in later life. Contributions to the etiological understanding of the anxiety disorders will also be reviewed: changes in variables considered important in the etiology and maintenance of disorder can be examined in synchronicity with changes in symptomatology following intervention. An examination of potential developmental predictors of treatment outcome will also contribute to this review, with a focus on the limitations of the current research in gaining a complete understanding of the relationship between developmental level and outcome. Directions regarding future research in the study of interventions for child and adolescent anxiety disorders will be discussed with the aim of promoting further communication between intervention research and the field of developmental psychopathology. PMID- 12549706 TI - Interventions for foster parents: implications for developmental theory. AB - All children who enter foster care have experienced disruptions in their relationships with caregivers, and many have experienced maltreatment. Studying the effects of these adverse early experiences can inform developmental theory. In particular, insight can be provided regarding sensitive periods in the development of attachment and self-regulatory capabilities. The quality of surrogate caregivers varies as a function of both the intervention services provided and foster parent characteristics. Studying the effects of foster parent quality can suggest which aspects of child functioning are more or less canalized at various developmental periods. This paper considers salient developmental issues of infancy, preschool years, middle childhood, and adolescence and examines ways in which these issues may present special difficulties for foster children. Across development, foster care is associated with difficulties regulating behaviors, emotions, and physiology. Thus, conditions associated with foster care placement (e.g., disruptions in care, maltreatment) appear to affect very basic and fundamental regulatory processes. Interventions have been designed that target developmentally specific manifestations of regulatory difficulties. Although the literature regarding evidence-based interventions for foster parents is quite limited, preliminary findings provide some evidence that nurturing, responsive care can serve to partially remediate early deficits. These findings suggest that stable and nonfrightening care is essential for normal development. Nonetheless, even in the case of quite adverse early experience that results in problematic child outcomes, there is some evidence that the development of many systems remains relatively plastic. PMID- 12549707 TI - Where education and mental health meet: developmental prevention and early intervention in schools. AB - There is growing recognition that traditional strategies, such as pullout special education programs and on-site casework and guidance, are no longer sufficient to address the staggering needs of a growing number of troubled children entering U.S. schools. This paper introduces a school-based prevention and intervention method for young adolescents called Responsive Advocacy for Life and Learning in Youth (RALLY). Prevention practitioners, a new role developed by the program, work in classrooms and after-school settings to provide nonstigmatizing support to students. Using a three-tiered prevention model, practitioners integrate a mental health and educational focus to foster students' academic, social, and emotional success. The intervention involves all children of an age cohort in middle schools. The practitioners are developmental specialists who create relationships with youth, teachers, and families in high-risk environments and serve triage functions to existing community and health care institutions. The RALLY intervention builds on normative developmental and developmental psychopathology theory, especially a risk and resilience framework. The paper describes the principles behind the practice and how the work in this field has created innovations in theory and a new impetus for research. PMID- 12549708 TI - The relative efficacy of two interventions in altering maltreated preschool children's representational models: implications for attachment theory. AB - A narrative story-stem task was used to evaluate the efficacy of two competing, developmentally informed preventive interventions for maltreated preschoolers and their mothers designed to modify children's internal representations of self and of self in relation to other. One hundred and twenty-two mothers and their preschoolers (87 maltreated and 35 nonmaltreated) served as participants. Maltreating families were randomly assigned to either the preschooler-parent psychotherapy (PPP, n = 23), psychoeducational home visitation (PHV, n = 34), or community standard (CS, n = 30) intervention group at baseline. Thirty-five nonmaltreating (NC) families served as comparisons. Narratives were administered to children at baseline and at the postintervention evaluation. Children in the PPP intervention evidenced more of a decline in maladaptive maternal representations over time than PHV and CS children and displayed a greater decrease in negative self-representations than CS, PHV, and NC children. Also, the mother-child relationship expectations of PPP children became more positive over the course of the intervention, as compared to NC and PHV participants. These results suggest that an attachment-theory informed model of intervention (PPP) is more effective at improving representations of self and of caregivers than is a didactic model of intervention directed at parenting skills. Findings are discussed with respect to their implications for developmental theory, with a specific focus on attachment theory and internal working models of relationships. PMID- 12549709 TI - A longitudinal-experimental approach to testing theories of antisocial behavior development. AB - A longitudinal study with a nested preventive intervention was used to test five hypotheses generated from developmental theories of antisocial behavior. The longitudinal study followed 909 boys from their kindergarten year up to 17 years of age. The randomized multimodal preventive intervention targeted a subsample of boys who were rated disruptive by their kindergarten teacher. Semiparametric analyses of developmental trajectories for self-reported physical aggression, vandalism, and theft identified more types of trajectories than expected from recent theoretical models. Also, these trajectories did not confirm theoretical models, which suggest a general increase of antisocial behavior during adolescence. The majority of boys were on either a low-level antisocial behavior trajectory or a declining trajectory. Less than 6% appeared to follow a trajectory of chronic antisocial behavior. Comparisons between disruptive and nondisruptive kindergarten boys confirmed the hypothesis that disruptive preschool children are at higher risk of following trajectories of frequent antisocial behavior. Comparisons between treated and untreated disruptive boys confirmed that an intensive preventive intervention between 7 and 9 years of age, which included parent training and social skills training, could change the long term developmental trajectories of physical aggression, vandalism, and theft for disruptive kindergarten boys in low socioeconomic areas. The results suggest that trajectories of violent behavior can be deflected by interventions that do not specifically target the physiological deficits that are often hypothesized to be a causal factor. The value of longitudinal-experimental studies from early childhood onward is discussed. PMID- 12549710 TI - Using the Fast Track randomized prevention trial to test the early-starter model of the development of serious conduct problems. AB - The Fast Track prevention trial was used to test hypotheses from the Early Starter Model of the development of chronic conduct problems. We randomly assigned 891 high-risk first-grade boys and girls (51% African American) to receive the long-term Fast Track prevention or not. After 4 years, outcomes were assessed through teacher ratings, parent ratings, peer nominations, and child self-report. Positive effects of assignment to intervention were evident in teacher and parent ratings of conduct problems, peer social preference scores, and association with deviant peers. Assessments of proximal goals of intervention (e.g., hostile attributional bias, problem-solving skill, harsh parental discipline, aggressive and prosocial behavior at home and school) collected after grade 3 were found to partially mediate these effects. The findings are interpreted as consistent with developmental theory. PMID- 12549711 TI - Contextual social-cognitive mediators and child outcome: a test of the theoretical model in the Coping Power program. AB - This study tests the contextual social-cognitive model, which has served as the basis for the Coping Power program, an indicated preventive intervention with at risk preadolescent boys at the time of transition from elementary to middle school. The contextual social-cognitive model assumes that aggressive children have distortions in their social-cognitive appraisals and deficiencies in their social problem solving skills and that their parents have deficiencies in their parenting behaviors. To test this model, boys were identified as being at risk on the basis of fourth grade and fifth grade teachers' ratings of children's aggressive and disruptive behaviors, and interventions were delivered at the end of elementary school and the beginning of middle school. The intervention effect on delinquency, substance use, and school behavior outcomes was at least partially mediated through intervention-produced changes in child and parent variables that were targets for the intervention. These analyses provided unique support for the assumptions in the contextual social-cognitive model that changes in these mediating processes, even among high-risk boys, can have a meaningful impact on later negative outcomes. PMID- 12549712 TI - Pragmatism in modeling peer influence: dynamics, outcomes, and change processes. AB - This paper reviews the model-building approach to developmental psychopathology as it applies to understanding the influence of peers on adolescent problem behavior. Considerable longitudinal evidence suggests that deviant peer involvement is prognostic of escalations in various forms of problem behavior, including substance use, delinquent behavior, and violence. Most convincing, however, are data from randomized intervention trials that show alarming iatrogenic effects for aggregating high-risk peers. This paper shows that intervention research can also clarify the role of parents in reducing deviant peer involvement. Results from a 4-year prevention trial are reported for a community sample of multiethnic middle school youth. Random assignment to family centered services in a school Family Resource center produced reduced growth in deviant peer involvement. The extensiveness of the parenting intervention was correlated with reduced growth in deviant peer involvement, especially among high risk youth. Findings are discussed with respect to developmental theory and the design of future intervention trials that clarify the role of adults in structuring the emerging adolescents' world of peers. PMID- 12549713 TI - Neuronal networks in emotions. PMID- 12549714 TI - "Ottorino Rossi" Award 2002. PMID- 12549715 TI - A molecular approach to retinal neural networks. AB - Our understanding of the computations carried out by neural networks in the central nervous system is limited by our incomplete knowledge of the diversity of cell types and the multiplicity of their functions. In the retina, over fifty cell types encode the spatial, temporal, and chromatic parameters of the incoming light stimuli to generate the messages of action potentials that travel to the brain along the fibers of the optic nerve. We have combined molecular techniques with microscopy and electrophysiology to study a rare cell type in the retina, the dopaminergic amacrine (interplexiform cells). These neurons were labeled in transgenic mice with human placental alkaline phosphatase so that they could be identified in vitro for patch-clamping and analysis of gene expression by single cell RT-PCR and cDNA array profiling. We have thus demonstrated that the retinal dopaminergic neurons spontaneously generate action potentials in a rhythmic fashion, release dopamine over their entire surface, establish GABAergic synapses and contain proteins whose existence went undetected with more conventional techniques. PMID- 12549716 TI - Reference fields in phantom tooth pain as a marker for remapping in the facial territory. AB - Six patients with chronic phantom tooth pain were studied for the presence of reference fields for their phantom sensation. In five of them, pain or dysesthesia in the affected oral structures was elicited by thermal or mechanical stimulation of areas that were well separate from these structures. However, a relation of topographical proximity between the stimulated areas and the areas of reference could be traced in the sensory maps. Therefore, denervation of small structures with coarse sensitivity can yield the plastic changes that have previously been described for larger deafferentations of areas endowed with finer discriminative capacity. PMID- 12549717 TI - Sympathetic skin response evoked by laser skin stimulation. AB - The objective of this study was to evoke sympathetic skin responses (SSRs) in healthy subjects using laser stimulation and to compare these responses with those induced by conventional electrical stimuli. Twenty healthy subjects were investigated. SSRs were obtained using electrical and laser stimuli delivered to the wrist controlateral to the recording site. The sympathetic sudomotor conduction velocity (SSFCV) was measured in 8 subjects by simultaneously recording the SSR from the hand and the axilla. The latency (L) of the laser induced SSR (ISSR) was significantly longer than that of the electrically-evoked SSR (eSSR) (mean ISSRL= 1.7 +/- 0.145 ms, mean eSSRL= 1.56 +/- 0.14 ms, p<0.05). The amplitude (A) of the ISSR was lower than the eSSR amplitude (mean ISSRA = 1.31 +/- 0.26 mV, mean eSSRA = 2.59 +/- 0.49 mV, p<0.05). No significant difference between the ISSR and eSSR was observed in either the SSFCV or the variability and reproducibility parameters. Our findings show that SSRs can easily be induced by laser stimuli and that this method shares the technical limitations of conventional eSSRs. PMID- 12549718 TI - Trigeminal evoked potentials and sensory deficits in atypical facial pain--a comparison with results in trigeminal neuralgia. AB - Trigeminal evoked potentials (TEPs) and sensory deficits in eighty-three patients admitted for first surgical treatment of facial pain were retrospectively analysed. Thirty-seven patients suffered from trigeminal neuralgia (TN), 10 from symptomatic TN (sTN), and 36 from atypical facial pain (AFP). Eighteen percent of the TN patients reported sensory deficits on the pain side, but 35% had delayed ipsilateral N13 waves. Of the sTN patients, 60% had either sensory deficits or a pathological corneal reflex and 62.5% a pathological N13. Of the AFP patients, 61% complained of sensory deficits, but only 31% had a pathological N13. The percentage of pathological P19 waves was slightly lower (20%, 50%, and 11%, respectively), but showed a similar trend. Normal TEPs were found even in the presence of a sensory deficit (reported only in the AFP group). These findings may add weight to the hypothesis of underlying psychiatric disorders in AFP. PMID- 12549719 TI - A reliability study of impairment and disability scales for myasthenia gravis patients. AB - The authors developed two scales to be adopted for the evaluation of myasthenia gravis (MG) patients. The first scale (MG impairment scale) is based on objective patient evaluation and on patients' responses to standardized questions relating to the functioning of specific muscle groups. It consists of 13 items exploring strength and 10 items exploring fatigability. The second scale (MG disability scale) evaluates disability in those everyday activities that are often impaired in MG patients. Test-retest reliability of each item and of the global score (sum of single item scores) was assessed by the weighted K statistic and by the intraclass correlation coefficient. Reliability was invariably 'substantial', and for single items 'almost perfect' for the MG impairment scale, and invariably 'almost perfect' for the MG disability scale. The internal structure of the MG impairment scale was explored by means of the principal component analysis. This analysis resulted in three main (rotated) factors, which loaded respectively onto 'ocular', 'spinal' and 'bulbar' functions. For these factors, we report factor score coefficients that can be used to compute single patients' scores, which in turn may be used in further analyses, particularly for follow-up studies. We also report the results of an analysis of the correlations between the two scales. The MG impairment and the MG disability scales are proposed for application in both clinical and research settings. PMID- 12549720 TI - Multiple sclerosis and parkinsonism: a case report. AB - Multiple sclerosis (MS) is a well-known disease characterized by the distribution of plaques in the periventricular and subcortical white matter. Although plaques can also be found in the striatum, pallidum and thalamus, extrapyramidal symptoms are very rare in MS. However, the association of MS and parkinsonism is still a controversial topic as it has not been established whether these two conditions occur coincidentally or causally. In the literature, eleven cases of parkinsonism associated with MS have been described. Here, we report a patient with clinically definite MS and signs of parkinsonism. Our patient had slow progressive bradykinesia, static tremor and bradymimia that were not associated with exacerbation or progression of the MS. This rare and interesting association of multiple sclerosis with parkinsonism is discussed in the light of literature reports. PMID- 12549722 TI - Patient information. Migraine treatments--what you can expect. PMID- 12549721 TI - Relieving migraine pain: sorting through the options. AB - Although triptans are a major advance in the treatment of migraine, the optimal approach for acute treatment involves a combination of lifestyle modifications, nonpharmacologic symptom relief, and drug therapy. PMID- 12549723 TI - Polycystic ovary syndrome: pathogenesis and treatment over the short and long term. AB - Although polycystic ovary syndrome (PCOS) is associated with hyperandrogenism and infertility early in life, it is a harbinger of a lifelong condition that can lead to serious sequelae such as endometrial or ovarian cancer, diabetes mellitus, and coronary artery disease. We review the pathophysiology, diagnosis, and treatment of this condition. PMID- 12549724 TI - Should everyone with a recent myocardial infarction receive a beta-blocker and an ACE inhibitor? PMID- 12549726 TI - Patient information. If your tuberculosis skin test is positive. PMID- 12549725 TI - New guidelines: what to do about an unexpected positive tuberculin skin test. AB - The 2000 recommendations for tuberculosis testing from the American Thoracic Society and the Centers for Disease Control and Prevention advocate a shift in focus from screening the general population to testing only patients at increased risk of developing tuberculosis. PMID- 12549727 TI - Thyrotoxicosis and the cardiovascular system: subtle but serious effects. AB - Thyrotoxicosis is associated with increased cardiovascular morbidity and mortality, primarily due to heart failure and thromboembolism. However, its signs and symptoms may be subtle and can easily be missed. Therefore, one should suspect thyrotoxicosis in patients with palpitations, exercise intolerance, dyspnea on exertion, and other cardiovascular signs. PMID- 12549728 TI - Alosetron (Lotronex) is back: should I use it to treat my patients with irritable bowel syndrome? PMID- 12549729 TI - A 62-year-old man with sudden onset of orthopnea and left shoulder weakness and pain. PMID- 12549730 TI - Modeling of dynamic cardiovascular responses during G-transition-induced orthostatic stress in pitch and roll rotations. AB - Dynamic and fuzzy models for a typical subject's cardiovascular response to the orthostatic stress have been developed based on experimental data. In our original study (Cheung et al., 1999), arterial blood pressure (BP) time-series data were obtained using a man-rated tilt table that applies gigahertz acceleration transitions from +0.861 Gz [head-up (HU)] to -0.707 G [head-down (HD)] and back to +0.861 Gz (HU) using either pitch or roll rotations (Cheung et al., 1999). G transitions of different duration and onset rates are common in fighter maneuvers. Based on these data, two types of predictive models have been developed in this paper: 1) second-order discrete-time models that predict BP dynamics during pitch and roll rotations and 2) fuzzy logic models that predict important variations in a subject's cardiovascular dynamics induced by HU-to-HD and HD-to-HU transitions. These two types of models assist in providing an operationally important predictive view on the characteristics of BP responses to orthostatic stress induced by pitch and roll rotations of a fighter jet pilot. The new models are being currently utilized in the design of operational recommendations for more G-tolerant operational flight regimes (e.g., split-S tactical maneuver) than the ones currently in use for modern combat aircraft. PMID- 12549731 TI - Rigorous Green's function formulation for transmembrane potential induced along a 3-D infinite cylindrical cell. AB - The quasi-static electromagnetic field interaction with three-dimensional infinite-cylindrical cell is investigated for both intracellular (IPS) and extracellular (EPS) current point-source excitation. The induced transmembrane potential (TMP), expressed conventionally via Green's function, may alternatively be expanded into a faster-converging representation using a complex contour integration, consisting of an infinite-discrete set of exponentially decaying oscillating modes (corresponding to complex eigenvalues) and a continuous source mode convolution integral. The dominant contributions for both the IPS and EPS problems are obtained in simple closed-form expressions, including well documented special mathematical functions. In the IPS case, the dominant modal contribution (of order zero)--an exact solution of the well-known cable equation- is explicitly and analytically corrected by the imaginary part of its eigenvalue and the source-mode convolution contribution. However, the TMP along a fiber was shown to decay at infinity algebraically and not exponentially, as predicted by the classic cable equation solution. In the EPS case, the dominant contribution is expressed as a source-mode convolution integral. However, for a long EPS distance (e.g., >10 cable length constant) the order-one-modes involved in the convolution is not a solution of the cable equation. Only for shorter EPS distance should the cable equation solution (i.e., the order zero dominant mode) be included in addition to the modes of order one. For on-membrane EPS location, additional modes should be included as well. In view of our EPS result, we suggest that the cable equation modeling existing in the literature and related to functional electrical stimulation for EPS problems, should be critically reviewed and corrected. PMID- 12549732 TI - A method for quantifying atrial fibrillation organization based on wave morphology similarity. AB - A new method for quantifying the organization of single bipolar electrograms recorded in the human atria during atrial fibrillation (AF) is presented. The algorithm relies on the comparison between pairs of local activation waves (LAWs) to estimate their morphological similarity, and returns a regularity index (rho) which measures the extent of repetitiveness over time of the detected activations. The database consisted of endocardial data from a multipolar basket catheter during AF and intraatrial recordings during atrial flutter. The index showed maximum regularity (rho = 1) for all atrial flutter episodes and decreased significantly when increasing AF complexity as defined by Wells (type I: rho = 0.75 +/- 0.23; type II: rho = 0.35 +/- 0.11; type III: rho = 0.15 +/- 0.08; P < 0.01). The ability to distinguish different AF episodes was assessed by designing a classification scheme based on a minimum distance analysis, obtaining an accuracy of 85.5%. The algorithm was able to discriminate among AF types even in presence of few depolarizations as no significant rho changes were observed by reducing the signal length down to include five LAWs. Finally, the capability to detect transient instances of AF complexity and to map the local regularity over the atrial surface was addressed by the dynamic and multisite evaluation of rho, suggesting that our algorithm could improve the understanding of AF mechanisms and become useful for its clinical treatment. PMID- 12549733 TI - A resampling approach to estimate the stability of one-dimensional or multidimensional independent components. AB - When applying unsupervised learning techniques in biomedical data analysis, a key question is whether the estimated parameters of the studied system are reliable. In other words, can we assess the quality of the result produced by our learning technique? We propose resampling methods to tackle this question and illustrate their usefulness for blind-source separation (BSS). We demonstrate that our proposed reliability estimation can be used to discover stable one-dimensional or multidimensional independent components, to choose the appropriate BSS-model, to enhance significantly the separation performance, and, most importantly, to flag components that carry physical meaning. Application to different biomedical testbed data sets (magnetoencephalography (MEG)/electrocardiography (ECG) recordings) underline the usefulness of our approach. PMID- 12549734 TI - Automated detection and elimination of periodic ECG artifacts in EEG using the energy interval histogram method. AB - An automated method for electrocardiogram (ECG)-artifact detection and elimination is proposed for application to a single-channel electroencephalogram (EEG) without a separate ECG channel for reference. The method is based on three characteristics of ECG artifacts: the spike-like property, the periodicity and the lack of correlation with the EEG. The method involves a two-step process: ECG artifact detection using the energy interval histogram (EIH) method and ECG artifact elimination using a modification of ensemble average subtraction. We applied a smoothed nonlinear energy operator to the contaminated EEG, which significantly emphasized the ECG artifacts compared with the background EEG. The EIH method was initially proposed to estimate the rate of false positives (FPs) and false negatives (FNs) that were necessary to determine the optimal threshold for the detection of the ECG artifact. As a postprocessing step, we used two types of threshold adjusting algorithms that were based on the periodicity of the ECG R-peaks. The technique was applied to four whole-night sleep EEG recordings from four subjects with severe obstructive sleep apnea syndrome, from which a total of 132878 heartbeats were monitored over 31.8 h. We found that ECG artifacts were successfully detected and eliminated with FP = 0.017 and FN = 0.074 for the epochs where the elimination process is necessarily required. PMID- 12549735 TI - Performance of an MEG adaptive-beamformer technique in the presence of correlated neural activities: effects on signal intensity and time-course estimates. AB - The influence of temporarily correlated source activities on neuromagnetic reconstruction by adaptive beamformer techniques was investigated. It is known that the spatial filter weight of an adaptive beamformer cannot perfectly block correlated signals. This causes two major influences on the reconstruction results: time course distortions and reductions in reconstructed signal intensities. Our theoretical analysis and numerical experiments both showed that the reduction in signal intensity for sources with a medium degree of correlation is small. The time-course distortion for such sources, however, may be discernible. Our analysis also showed that the magnitude correlation coefficient between two correlated sources can be accurately estimated by using the beamformer outputs. A method of retrieving the original time courses using estimated correlation coefficients was developed. Our numerical experiments demonstrated that reasonably accurate time courses can be retrieved from considerably distorted time courses even when the signal-to-noise ratio is low. PMID- 12549736 TI - Instantaneous parameter estimation in cardiovascular time series by harmonic and time-frequency analysis. AB - Time-frequency distributions, such as smoothed pseudo Wigner-Ville distribution (SPWVD), complex demodulation (CDM), and provide useful time-varying spectral parameter estimators. However, each of these methods has limitations that a joint utilization could largely reduce, due to their interesting complementary features. The aim of this paper is to validate the joint SPWVD-CDM method on synthetic and real cardiovascular time series with normal and reduced variability such as in autonomic blockade or autonomic deficiency. We propose two indexes related to the noise present in the signal and to the dispersion of the power spectrum in order to validate instantaneous parameter estimation. In the low frequency band, the interpretation of the instantaneous frequency and phase of cardiovascular time-series should be discarded in many real-life situations. Conversely, in the high frequency band, under paced breathing, the reliability of the instantaneous parameters is demonstrated even in conditions of reduced cardiovascular variability. PMID- 12549737 TI - A multistage, multimethod approach for automatic detection and classification of epileptiform EEG. AB - An efficient system for detection of epileptic activity in ambulatory electroencephalogram (EEG) must be sensitive to abnormalities while keeping the false-detection rate to a low level. Such requirements could be fulfilled neither by single stage nor by simple method strategy, due to the extreme variety of EEG morphologies and frequency of artifacts. The present study proposes a robust system that combines multiple signal-processing methods in a multistage scheme, integrating adaptive filtering, wavelet transform, artificial neural network, and expert system. The system consists of two main stages: a preliminary screening stage in which data are reduced significantly, followed by an analytical stage. Unlike most systems that merely focus on sharp transients, our system also takes into account slow waves. A nonlinear filter for separation of nonstationary and stationary EEG components is also developed in this paper. The system was evaluated on testing data from 81 patients, totaling more than 800 hours of recordings. 90.0% of the epileptic events were correctly detected. The detection rate of sharp transients was 98.0% and overall false-detection rate was 6.1%. We conclude that our system has good performance in detecting epileptiform activities and the multistage multimethod approach is an appropriate way of solving this problem. PMID- 12549738 TI - Nonlinear model for offline correction of pulmonary waveform generators. AB - Pulmonary waveform generators consisting of motor-driven piston pumps are frequently used to test respiratory-function equipment such as spirometers and peak expiratory flow (PEF) meters. Gas compression within these generators can produce significant distortion of the output flow-time profile. A nonlinear model of the generator was developed along with a method to compensate for gas compression when testing pulmonary function equipment. The model and correction procedure were tested on an Assess Full Range PEF meter and a Micro DiaryCard PEF meter. The tests were performed using the 26 American Thoracic Society standard flow-time waveforms as the target flow profiles. Without correction, the pump loaded with the higher resistance Assess meter resulted in ten waveforms having a mean square error (MSE) higher than 0.001 L2/s2. Correction of the pump for these ten waveforms resulted in a mean decrease in MSE of 87.0%. When loaded with the Micro DiaryCard meter, the uncorrected pump outputs included six waveforms with MSE higher than 0.001 L2/s2. Pump corrections for these six waveforms resulted in a mean decrease in MSE of 58.4%. PMID- 12549739 TI - In vitro electrical properties for iridium oxide versus titanium nitride stimulating electrodes. AB - Stimulating electrode materials must be capable of supplying high-density electrical charge to effectively activate neural tissue. Platinum is the most commonly used material for neural stimulation. Two other materials have been considered: iridium oxide and titanium nitride. This study directly compared the electrical characteristics of iridium oxide and titanium nitride by fabricating silicon substrate probes that differed only in the material used to form the electrode. Electrochemical measurements indicated that iridium oxide had lower impedance and a higher charge storage capacity than titanium nitride, suggesting better performance as a stimulating electrode. Direct measurement of the electrode potential in response to a biphasic current pulse confirmed that iridium oxide uses less voltage to transfer the same amount of charge, therefore using less power. The charge injection limit for titanium nitride was 0.87 mC/cm2, contradicting other reports estimating that titanium nitride was capable of injecting 22 mC/cm2. Iridium oxide charge storage was 4 mC/cm2, which is comparable to other published values for iridium oxide. Electrode efficiency will lead to an overall more efficient and effective device. PMID- 12549740 TI - Modeled channel distributions explain extracellular recordings from cultured neurons sealed to microelectrodes. AB - Amplitudes and shapes of extracellular recordings from single neurons cultured on a substrate embedded microelectrode depend not only on the volume conducting properties of the neuron-electrode interface, but might also depend on the distribution of voltage-sensitive channels over the neuronal membrane. In this paper, finite-element modeling is used to quantify the effect of these channel distributions on the neuron-electrode contact. Slight accumulation or depletion of voltage-sensitive channels in the sealing membrane of the neuron results in various shapes and amplitudes of simulated extracellular recordings. However, estimation of channel-specific accumulation factors from extracellular recordings can be obstructed by co-occuring ion currents and defect sealing. Experimental data from cultured neuron-electrode interfaces suggest depletion of sodium channels and accumulation of potassium channels. PMID- 12549741 TI - Extracellular stimulation window explained by a geometry-based model of the neuron-electrode contact. AB - Extracellular stimulation of single cultured neurons which are completely sealing a microelectrode is usually performed using anodic or biphasic currents of at least 200 nA. However, recently obtained experimental data demonstrate the possibility to stimulate a neuron using cathodic current pulses with less amplitude. Also, a stimulation window is observed. These findings can be explained by a finite-element model which permits geometry-based electrical representation of the neuron-electrode interface and can be used to explore the required conditions for extracellular stimulation in detail. Modulation of the voltage sensitive channels in the sealing part of the membrane appears to be the key to successful cathodic stimulation. Furthermore, the upper limit of the stimulation window can be explained as a normal consequence of the neuronal membrane electrophysiology. PMID- 12549742 TI - Electrical coupling of single cardiac rat myocytes to field-effect and bipolar transistors. AB - A novel bipolar transistor for extracellular recording the electrical activity of biological cells is presented, and the electrical behavior compared with the field-effect transistor (FET). Electrical coupling is examined between single cells separated from the heart of adults rats (cardiac myocytes) and both types of transistors. To initiate a local extracellular voltage, the cells are periodically stimulated by a patch pipette in voltage clamp and current clamp mode. The local extracellular voltage is measured by the planar integrated electronic sensors: the bipolar and the FET. The small signal transistor currents correspond to the local extracellular voltage. The two types of sensor transistors used here were developed and manufactured in the laboratory of our institute. The manufacturing process and the interfaces between myocytes and transistors are described. The recordings are interpreted by way of simulation based on the point-contact model and the single cardiac myocyte model. PMID- 12549743 TI - Dynamic impedance measurements during radio-frequency heating of cornea. AB - Hyperopia affects approximately 25% of the population. The aim of different heating modalities for the treatment of hyperopia is to steepen the central curvature of the cornea. Conductive keratoplasty (CK) involves the placement of radio-frequency (RF) lesions around a 7-mm-diameter ring concentric with the pupil of the eye. Dynamics of lesion formation during CK depend on corneal electrical impedance, which is expected to change during each 600-ms-long macropulse. The purpose of this study was to measure impedance dynamics during CK. RF lesions were made in in vitro porcine eyes at different power settings. Voltage and current measurements were acquired using a high-speed computer-based data acquisition system. Root-mean-square voltages (VRMS) and curre (IRMS) were calculated for each micropulse, and impedance was determined by calculating the quotient VRMS/IRMS. Initial corneal impedance in vitro was approximately 2000 ohms. During the macropulse, impedance decreased initially due to increased mobility of conductive ions. At higher power settings (e.g., > 70%, or maximum peak-to-peak voltage of 233 V), impedance increased after the initial decrease, indicative of local water vaporization and/or tissue coagulation. Preliminary impedance data obtained for in vivo porcine eyes were similar in magnitude to the in vitro values. PMID- 12549744 TI - Automatic estimation of the corneal limbus in videokeratoscopy. AB - An algorithm for estimating the corneal limbus from videokeratoscopic images is proposed. After the image is transformed to a polar grid, a novel edge-detection procedure, suitable for the detection of the soft edge produced by the limbus, is used to locate the limbus. Outliers due to the eyelids, eyelashes, and videokeratoscopic rings are removed by taking advantage of the approximate circularity of the cornea. An ellipse which minimizes the sum of the squared algebraic errors is fitted to the remaining edge points. Comparisons between the proposed algorithm, a manual computer-based technique and an algorithm which uses conventional edge-detection techniques demonstrate the accuracy of the proposed algorithm. PMID- 12549745 TI - An explant model for the investigation of skin adaptation to mechanical stress. AB - A test apparatus was developed to investigate the effects of mechanical stress application on collagen remodeling in skin. The system maintained a 4.5-cm x 5.5 cm skin explant at an air interface with controlled temperature, relative humidity (RH), and carbon dioxide concentration [CO2] while allowing controlled compressive and shear forces to be applied to the skin surface. For environmental control, a custom-designed flow system under Proportional-Integral-Derivative (PID) control was used. Evaluation tests demonstrated that the system maintained air above the explant at a temperature within 1 degrees C of the 37.5 degrees C set point, RH within 5% of the user-specified set point (range of 5% to 95%), and [CO2] within 1% of the 5% [CO2] set point. Least-squares errors in cyclic compressive and shear forces (0- to 20-Hz bandwidth) delivered to the explant were 0.9% and 2.8%, respectively, of user-specified values. Pig skin samples cyclically stressed for 1 hr/day for 3 days with either compressive force only or a combined compressive and shear force had significantly smaller collagen fibril densities compared with an unstressed control, a result consistent with in vivo test data. Collagen fibril diameters were significantly larger for stressed versus control for some of the samples, but the changes were not as substantial as from in vivo testing. This result may have been due to the shorter study duration in vitro (3 d versus 20 d in vivo). The system allows insight into the mechanisms of skin adaptation to mechanical stress to be investigated on a cellular and molecular level, potentially leading to therapies to encourage adaptation in at-risk patients. PMID- 12549746 TI - Flow-rate measurements and models for colloid and crystalloid flows in central and peripheral venous line infusion systems. AB - The resuscitation fluids, including crystalloids and colloids, were tested in an experimental module with 16-gauge central and peripheral catheters. Infusion pressures were ranged from the gravity driving 10 kPa (75 mmHg) to the pressurized driving 50 kPa (375 mmHg). The experiment results were correlated to obtain the empirical friction factors and the loss factors for the components commonly used in a fluid resuscitation system. The modified Bernoulli equation with the correlated friction factors and loss factors for the components were used to evaluate the pressure and flow relationship in the fluid resuscitation system with peripheral and central catheters. Fair agreements were observed from the comparison of the predictions of the total driving pressure and test results from water, crystalloid and colloid solutions. The modified Bernoulli equation is, therefore, applicable to evaluate the pressure-flow relationship for efficient fluid resuscitations. The coefficients of flow (F) and the square of flow (F2) for the binominal model are varied with the changes of geometry and size of the infusion components, fluid properties and the units of parameters. These coefficients for the fluids and catheters tested in this study were also listed for reference. PMID- 12549747 TI - Fiberglass needle electrodes for transmural cardiac mapping. AB - We developed a new method for fabricating plunge needle electrodes for use in cardiac mapping. The needles have 12 electrodes with 1-mm spacing, are 0.5 mm in diameter, and are fabricated from fiberglass reinforced epoxy. They are stiff enough to be easily inserted into beating hearts and durable enough to be reused many times. We found that these new needles elicit smaller, more quickly resolving injury potentials, and when inserted in a row with 2-mm spacing, disrupt ventricular fibrillation activation patterns less than traditional steel needles. PMID- 12549748 TI - Wave propagation in the atrial myocardium: dispersion properties in the normal state and before fibrillation. AB - The formalism of wave propagation in passive media is applied to the spread of the electrical excitation in the human atrial myocardium. From an analog of the classical dispersion dependence that is obtained by wavelet decomposition a precursor parameter is calculated that serves to predict fibrillation already before its onset. PMID- 12549749 TI - Reliability and validity of the Observational Gait Scale in children with spastic diplegia. AB - The aim of this study was to establish the reliability and validity of visual gait assessment in children with spastic diplegia, who were community or household ambulators, using a modified version of the Physicians Rating Scale, known as the Observational Gait Scale (OGS). Two clinicians viewed edited split screen video recordings of 20 children/adolescents (11 males, 9 females; mean age 12 years, range 6 to 21 years) made at the time of three-dimensional gait analysis (3-DGA). Walking ability in each child was scored at initial assessment and reassessed from the same videos three months later using the first seven sections of the OGS. Validity of the OGS score was determined by comparison with 3-DGA. The OGS was found to have acceptable interrater and intrarater reliability for knee and foot position in mid-stance, initial foot contact, and heel rise with weighted kappas (wk) ranging from 0.53 to 0.91 (intrarater) and 0.43 to 0.86 (interrater). Comparison with 3-DGA suggests that these sections might also have high validity(wk range 0.38-0.94). Base of support and hind foot position had lower interrater and intrarater reliabilities (wk 0.29 to 0.71 and wk 0.30 to 0.78 respectively) and were not easily validated by 3-DGA. PMID- 12549750 TI - Prevalence of reduced bone mass in children and adults with spastic quadriplegia. AB - This study was designed to test the hypothesis that non-ambulatory patients with spastic quadriplegia will have reduced bone mass which worsens with increasing age. Forty-eight patients (age 5 to 48 years, median age 15 years; 19 females and 29 males) were studied. Anticonvulsants were used in 29 patients (60.4%). Lumbar spine bone mineral density (LS-BMD) was markedly reduced compared with age-and sex-matched control individuals with a z score of -2.37 +/- 0.21. Twenty-eight (58%) had z scores of less than -2. A history of documented previous fracture was present in 19 patients (39%). Patients with a history of fracture had significantly lower (p = 0.05) LS-BMD z scores (-2.81 +/- 0.29) compared with those without a history of fracture (-2.11 +/- 0.26). Mean serum 25-OH vitamin D was 29.6 +/- 1.9ng/mL (normal 9 to 37.6ng/mL) with three patients having serum 25 OH vitamin D levels less than 15ng/mL. These findings indicate that BMD is markedly reduced in non-ambulatory children and adults with neuromuscular disease. Reductions in bone mass put them at greater risk for non-traumatic fractures. PMID- 12549751 TI - Effect of asphyxia on non-protein-bound iron and lipid peroxidation in newborn infants. AB - The effect of asphyxia on iron metabolism and lipid peroxidation in newborn infants with hypoxic-ischemic encephalopathy (HIE) was investigated. Non-protein bound iron (NPBI) and lipid peroxidation (thiobarbituric-acid-reactive species; TBARS) in plasma and hematological iron indices were measured in 15 healthy newborn infants (mean gestational age 39 04 weeks, SD 1); 15 asphyxiated infants without neurological abnormalities (AS-HIE; mean gestational age 38.8 weeks, SD 0.9); and 15 asphyxiated infants with neurological abnormalities (AS+HIE; mean gestational age 39.75 weeks, SD 1.4). Follow-up was performed at the age of 5 months. It was found that the detectable rates of NPBI in 10 of 15 of the AS-HIE group and 13 of 15 of the AS+HIE group were significantly higher than that of the control group (5 of 15; both p < 0.01). Plasma levels of TBARS in the control (9.20 micromol/L, SD 1.9) and AS-HIE infants (10.13 micromol/L, SD 2.7) were significantly lower than those of the AS+HIE group (13.42 micromol/L, SD 2.8). Serum iron, total iron binding capacity, and transferrin saturation in the AS+HIE group was higher than the corresponding values of the control and AS-HIE groups, although no statistical difference was found among them. At 5 months of age, all control and AS-HIE infants were neurologically normal, whether or not their NPBI was detectable. Of the 12 AS+HIE infants, four (all of whom had detectable NPBI) were neurologically impaired. The average Gross Development Quotient of AS+HIE infants was significantly lower than that of the control or AS-HIE groups (p < 0.01). Results showed that asphyxia could affect iron metabolism and lead to a significant increase in NPBI and lipid peroxidation in newborn infants with HIE, indicating that iron delocalization induced by asphyxia plays a role in the brain injury of asphyxiated infants. PMID- 12549752 TI - Long-term risks and benefits of a separate CSF access device with ventriculoperitoneal shunting in childhood hydrocephalus. AB - To determine the long-term risks and benefits of a separate CSF reservoir in the management of 52 children (23 males, 29 females) with shunted hydrocephalus, a retrospective study was performed comparing the use and complications after separate reservoir insertion, with a prereservoir control period. Median age at first shunt insertion was 1 month and median age at reservoir insertion was 2 years 6 months. Median follow-up for shunt with the additional reservoir was 19 years 1 month. There was no mortality due to shunt failure nor CNS infection, and there were significantly fewer episodes of ventriculitis (p < 0.01) and shunt blockage (p < 0.0001) compared with the prereservoir period. There was no hemiplegia, epilepsy, visual, nor cognitive loss from the additional reservoir. The reservoir was used for access in 344 attendances (mean 6.62 attendances per patient) for diagnosis or treatment of raised pressure or CNS infection. It was concluded that a separate CSF reservoir is useful in the long-term management of patients with shunted hydrocephalus and is without mortality or significant increased morbidity. PMID- 12549753 TI - Speech, language, and cognitive development in children with isolated sagittal synostosis. AB - This study investigated the occurrence, nature, and severity of speech, language, and cognitive impairment in 76 children (61 males, 15 females) with isolated sagittal synostosis (ISS) aged 9 months to 15 years 7 months. There was no increased prevalence of global cognitive impairment in the group but there was a high prevalence rate of speech and/or language impairment with 28 (37%) displaying impairment of whom 20 (71%) had moderate or severe impairments that fulfilled the criteria for specific impairments. Prevalence rates were only increased for children over two years of age. Expressive language impairment occurred most frequently. Raised intracranial pressure, peri-neonatal risk factors, otitis media, or being operated were not associated with impairment. Surgery at a later age and a family history of speech and language impairment were both associated with impairments but numbers were small. The findings suggest that children with ISS are at an increased risk of developing speech and language impairment. PMID- 12549754 TI - Behavioural problems in children and adolescents with spinal muscular atrophy and their siblings. AB - Spinal muscular atrophy (SMA) is a chronic illness characterized by loss of motor function. The aim of the study was to investigate behavioural adjustment in 96 children and adolescents with SMA (47 males, 49 females; mean age 11 years 2 months, range 6 to 18 years). Forty-five non-affected siblings (26 males, 19 females; mean age 11 years 6 months, range 6 to 18 years) and 59 normally developing children (33 males, 26 females; mean age 10 years 8 months, range 6 to 18 years) were recruited as control participants. Behavioural symptoms were measured with the Child Behaviour Checklist (CBCL) and disorders were assessed with a structured psychiatric interview (Kinder-DIPS). Of the patients with SMA, 12.5% fulfilled the criteria for an ICD-10 or DSM-IV diagnosis, with separation anxiety disorder being the most common diagnosis. The CBCL total score was in the clinical range for 11.5% of patients, 20% of the siblings, and 11.7% of the control children; the externalizing score rates were 2.1%, 22.2%, and 11.9% respectively; the internalizing score 18.9%, 24.4%, and 13.6% respectively. Comorbid psychopathology was not influenced by sex, IQ, nor severity of SMA, and only externalizing behaviour was correlated to age. In conclusion, children and adolescents with SMA are characterized by a low psychiatric comorbidity not different from control individuals. The group with the highest rate of behavioural problems and with the greatest need for intervention were the non affected siblings who had a two- to threefold higher rate of behavioural problems than the normative population. PMID- 12549755 TI - ADHD and epilepsy in childhood. AB - Attention-deficit-hyperactivity disorder (ADHD) has been associated with childhood epilepsy; prevalence figures have ranged from 8 to 77%, depending on the sample studied and the criteria used for diagnosis. In the general population the prevalence of ADHD is approximately 5%, with the majority of affected children having ADHD combined type. As part of a larger study of behavioral problems in children with epilepsy, we assessed 175 children (90 males, 85 females; age range 9 to 14 years, mean age was 11 years 10 months, SD 1 year 8 months) for evidence of ADHD. The children had at least a 6-month history of epilepsy. The primary caregiver completed both the Child Behavior Checklist (CBCL) and the Child Symptom Inventory-4 (CSI) or Adolescent Symptom Inventory-4 (ASI). On the CBCL, the mean attention problem T score was 64.6 (SD 10.5) for adolescents and 67.9 (SD 11.6) for children. On the CSI or ASI, 20 of 175 children met DSM-IV criteria for ADHD combined type; 42 of 175 had ADHD predominantly inattentive type; and 4 of 175 met criteria for ADHD predominantly hyperactive-impulsive type. There were significant correlations between the CBCL attention score and inattention (r = 0.68) and hyperactivity-impulsivity (r = 0.59). Sex, seizure type, and focus of seizure discharge were not predictors of symptoms of ADHD. Children with epilepsy are at risk for symptoms of ADHD. They differ from other samples of children with ADHD by the higher proportion of children with ADHD predominantly inattentive type and by an equal male: female ratio. PMID- 12549756 TI - Prosopagnosia in a preschool child with Asperger syndrome. AB - We investigated a male, aged 4 years 11 months, who fulfilled the criteria of Asperger syndrome). In addition to the typical pattern of autistic symptoms, psychological testing revealed prosopagnosia in tasks for face recognition and matching. Prosopagnosia was also present when he tried to identify the faces of his parents and himself in photographs whenever these were presented with photographs of other persons. Although impairment in reciprocal social interaction in individuals with Asperger syndrome is closely correlated to their impaired perceptional abilities in non-verbal communication, especially facial expression, overt prosopagnosia seems to be a rare neuropsychological symptom in persons with autistic disorders. PMID- 12549757 TI - Parental experience of participation in physical therapy for children with physical disabilities. PMID- 12549758 TI - Resilience: an account of the 2002 AACPDM meeting. PMID- 12549759 TI - 'Defining hypoxic-ischemic birth events'. PMID- 12549760 TI - Cancer chemoprevention. AB - Chemoprevention means prevention of cancer by administering chemical compounds. Ideal chemopreventive agents are non-toxic, inexpensive and can be taken orally. A large number of natural and synthetic compounds have cancer preventive properties in cell culture or animal model studies. With the demonstration of tamoxifen's ability to prevent breast cancer in women, the feasibility of chemoprevention in humans has now been firmly established. Current challenge is to build on this success and identify non-toxic chemopreventive agents. A promising area of research is clinical studies with botanicals, cancer preventive compounds in fruits, vegetables and other plants. PMID- 12549761 TI - Cancer chemoprevention by connexins. AB - Gap junctions, or connexons, are formed by connexin proteins and connect most cells in the body to form water-filled channels directly linking the cytoplasm. Among the molecules known to be transferred via junctions are cAMP, ATP, IP3 and glucose. Tumor cells are in general deficient in functional gap junctions either as a result of gene silencing, or failure to correctly process and assemble connexons. Tumor promoters inhibit function whereas certain cancer preventive agents increase junctional communication. When connexin expression in tumor cells is forced by introduction of exogenous genes or is increased by pharmacological agents, connexin expression reduces growth in suspension and growth as xenografts in nude mice. It is as yet unclear if in tumor cells these actions depend on junctional transfer of signal molecules or reflect some other function of these genes. Restoration of connexin function offers an exciting opportunity to delay tumor progression and inhibit metastasis. PMID- 12549762 TI - Micronutrients in cancer chemoprevention. AB - The selection of micronutrients, defined as essential and nonessential dietary components consumed in minute quantities, for testing in clinical chemoprevention trials is based on the totality of evidence arising from epidemiologic, in vitro, animal, and clinical studies. Those micronutrients that surface with chemopreventive potential, in terms of high efficacy and low toxicity, in early phase clinical studies are then candidates for large-scale, randomized clinical chemoprevention trials with cancer endpoints. Micronutrients currently being examined in National Cancer Institute (NCI)-sponsored phase I, II, or III chemoprevention trials for prostate, breast, and colon cancers include isoflavones, lycopene, selenized yeast, selenomethionine, selenium, vitamin E, perillyl alcohol, folic acid, vitamin D, calcium, and curcumin. The response to micronutrients may vary not only in magnitude but also in direction. This variation and response likely depend on individual genetic polymorphisms and/or interactions among dietary components that influence absorption, metabolism, or site of action. Research priorities include investigation of possible molecular targets for micronutrients and whether genetic and epigenetic events dictate direction and magnitude of the response. PMID- 12549763 TI - Botanicals in cancer chemoprevention. AB - Botanicals have been used for the treatment of various human diseases throughout history. In addition, botanicals play a role in disease prevention. For example, epidemiologic studies have suggested that a reduced risk of cancer is associated with high consumption of vegetables and fruits. Thus, the cancer chemopreventive potential of naturally occurring phytochemicals is of great interest. In this review, we discuss the cancer chemopreventive activity of cruciferous vegetables such as cabbage and broccoli, Allium vegetables such as garlic and onion, green tea, Citrus fruits, tomatoes, berries, ginger and ginseng, as well as some medicinal plants. In addition, methods for the discovery of active compounds from plant sources are described. Several lead compounds, such as brassinin (from cruciferous vegetables like Chinese cabbage), sulforaphane (from broccoli) and its analog sulforamate, withanolides (from tomatillos), and resveratrol (from grapes and peanuts among other foods), are in preclinical or clinical trials for cancer chemoprevention. Phytochemicals of these types have great potential in the fight against human cancer, and a variety of delivery methods are available as a result of their occurrence in nature. PMID- 12549764 TI - Carotenoids in cancer chemoprevention. AB - Various natural carotenoids, besides beta-carotene, were proven to have anticarcinogenic activity, and some of them showed more potent activity than beta carotene. Thus, these carotenoids (alpha-carotene, lutein, zeaxanthin, lycopene, beta-cryptoxanthin, fucoxanthin, astaxanthin, capsanthin, crocetin and phytoene), as well as beta-carotene, may be useful for cancer prevention. In the case of phytoene, the concept of 'bio-chemoprevention', which means biotechnology assisted method for cancerchemoprevention, may be applicable. In fact, establishment of mammalian cells producing phytoene was succeeded by the introduction of crtB gene, which encodes phytoene synthase, and these cells were proven to acquire the resistance against carcinogenesis. Antioxidative phytoene containing animal foods may be classified as a novel type of functional food, which has the preventive activity against carcinogenesis, as well as the ability to reduce the accumulation of oxidative damages, which are hazardous for human health. PMID- 12549765 TI - Mechanisms of cancer chemoprevention by soy isoflavone genistein. AB - Diet has been implicated to play an important role in cancers. Epidemiological studies have revealed that Asians, who consume a traditional diet high in soy products, have relatively low incidences of breast and prostate cancers, while the incidences are much higher in the Western world. Asians who immigrate to the United States and adopt a Western diet are at higher risks of breast and prostate cancers. Soy isoflavones have received much attention as dietary components having an important role in reducing breast and prostate cancers. Genistein, one of the predominant soy isoflavones, has been shown to inhibit the growth of cancer cells through the modulation of genes that are related to the homeostatic control of cell cycle and apoptosis. It has been found that genistein inhibits the activation of the nuclear transcription factor, NF-kappaB and Akt signaling pathway, both of which are known to maintain a balance between cell survival and programmed cell death (apoptosis). Genistein is known to have anti-oxidant property, and commonly known as phytoestrogen, which targets estrogen and androgen-mediated signaling pathway in the processes of carcinogenesis. Moreover, genistein is also found to be a potent inhibitor of angiogenesis and metastasis. Hence, significant advances have been made, both by in vitro and in vivo studies showing that genistein is a promising agent for cancer chemoprevention and/or treatment. PMID- 12549766 TI - New concepts in selenium chemoprevention. AB - This article highlights some recent advances in selenium cancer chemoprevention research. It has been well documented that the chemical transformation of selenium to a monomethylated metabolite is an important step in achieving cancer prevention. Studies with the rat mammary carcinogenesis model suggested that methylselenocysteine (MSC), a good precursor for generating methylselenol endogenously, is able to block clonal expansion of premalignant lesions in the mammary gland. This finding supports the notion that selenium intervenes at an early stage of carcinogenesis. In addition to decreasing cell proliferation of the transformed colonies in vivo, MSC also enhances apoptosis. These same cellular responses are replicated with human premalignant breast cells grown in culture. cDNA microarray analysis indicated that selenium affects a multitude of molecular targets. Based on this information, a number of signaling pathways are proposed that could potentially provide insight into how selenium might block cell cycle progression and induce cell death. PMID- 12549767 TI - Zinc in cancer prevention. AB - Epidemiologic studies suggest that zinc deficiency may be associated with increased risk of cancer. However, few studies have been conducted with zinc supplementation in animals and humans. Most previous intervention studies have used zinc in combination with several other micronutrients, which make them difficult to interpret. Zinc supplementation is associated with decreased oxidative stress and improved immune function, which may be among the possible mechanisms for its cancer preventive activity. Preclinical and clinical studies need to investigate modulation of genetic and epigenetic pathways of carcinogenesis by zinc. PMID- 12549768 TI - Chemoprevention of prostate cancer: current status and future directions. AB - Prostate cancer chemoprevention can be described as the administration of natural products and pharmaceutical agents that inhibit one or more steps in the natural history of prostatic carcinogenesis. The principle components of the chemoprevention strategy are closely connected to this natural history and include: (a) agents and their molecular targets; (b) strategic intermediate endpoint biomarkers (IEBs) and their critical pathways; (c) cohorts identified by genetic and acquired risk factors and (d) efficient designs that combine these elements into a cohesive clinical trial. The primary goal is to find effective noncytotoxic agents that modulate the promotion and progression from normal epithelium to dysplasia to high-grade prostatic intraepithelial neoplasia (HGPIN) to locally invasive cancer and metastatic disease. Another important target for chemoprevention is to modulate progression to clinically aggressive disease and to maintain an androgen-sensitive clinical state and delay the emergence of androgen resistance. There is a rationale for use of antiandrogens as the lead class, e.g., 5 alpha receptor inhibitors (5ARI), for chemoprevention of prostate cancer. Nevertheless, the desire to improve the therapeutic index, achieve synergy (5ARI may have only modest anticancer effects) and prevent the emergence of drug (androgen) resistance provide incentives for developing other effective agents and combinations. The availability of more than a dozen classes of noncytotoxic pharmaceutical and natural products already in clinical development create many opportunities for rational combination therapy. Several agent classes have a pharmacodynamic basis for combination with antiandrogens including antiproliferatives, selective estrogen receptor modulators (SERMs), proapoptotic antioxidant micronutrients and selective cyclo-oxygenase (COX)-2 inhibitors. Many other rational pharmacodynamic combinations without antiandrogens are feasible. It is anticipated that in the future, a selective COX-2 inhibitor may be combined with other agent classes such as proapoptotic antioxidant micronutrients, receptor tyrosine kinase modulators, antiangiogenic modulators, antiproliferative/differentiating agents, NFkappaB modulators, IGF-1 modulators and other novel proapototic nonsteroidal drugs. A novel target for rational combinations is the hypermethylation of GST-PI leading to functional silencing of this key anticarcinogen defense enzyme in precursors (HGPIN) and prostate cancer. Factorial designs are well suited for evaluating the individual and combined effects of each agent in a single trial design. There are a number of moderate to high-risk cohorts and clinical models of primary and secondary prevention that can be employed in both short-term developmental (translational) trials for proof of biologic activity and in intermediate sized longer-term chemoprevention trials for proof of efficacy against prostate cancer. Strategic IEBs are needed to more efficiently monitor short-term biologic activity and validate efficacy. The emergence of new powerful tools such as gene chip cDNA microarrays for multiplex gene expression profiling and proteomic analysis of tissue based and secreted proteins will accelerate the identification of new molecular targets, strategic endpoints, cohorts at risk and the design of rational combination trials. PMID- 12549769 TI - Chemoprevention of breast cancer: current and future prospects. AB - The groundwork for making the concept of breast cancer chemoprevention a clinical reality began over a century ago. Although tamoxifen's first clinical use was for the treatment of breast cancer, the earliest animal studies with the drug provided the scientific basis for chemoprevention. The extensive clinical experience, safety and laboratory data have made tamoxifen the current standard of-care for the prevention of breast cancer in women at elevated risk. The STAR trial will address the value of raloxifene as a chemopreventative in postmenopausal women. Results will be available by 2005. Newer compounds are under development which hold the promise of expanded efficacy and narrower side effect profile. These compounds will function as multifunctional medicines and will hold the promise of preventing breast and endometrial cancer, while providing the beneficial effects of preventing osteoporosis and coronary heart disease. PMID- 12549770 TI - Chemoprevention of colon cancer: current status and future prospects. AB - Colorectal cancer is an important public health problem in the western world. Although some progress has been made in the prevention and management of this disease, colon cancer still remains one of the most common types of epithelial malignancies in both genders and is essentially incurable when it reaches the most advanced stages. Given the substantial morbidity and mortality associated with colorectal malignancies and their treatment, cancer prevention in its many forms emerges as a very attractive approach. Colorectal cancer chemoprevention refers to the administration of natural or synthetic compounds to block, reverse, delay or prevent the development of invasive large bowel neoplasms. The ultimate goal of implementing a chemopreventive intervention in the general, or alternatively, in an at-risk population is to decrease the incidence rate of the specific cancer being targeted. This article reviews the present status of colorectal cancer chemoprevention. Current insights into the molecular and genetic models of human colorectal carcinogenesis, preclinical models for efficacy testing as well as into promising biomarkers for colorectal chemoprevention are provided. The developmental status of many promising agents is also discussed emphasizing the epidemiological evidence, preclinical information substantiating an anticarcinogenic effect, their postulated mechanism of action and the status of human clinical development. Our perspective of the future prospects in this scientific area is also provided and has been predicated primarily on the firm belief that the proper integration of advances in the biology of colon carcinogenesis, experimental therapeutics and clinical trial methodology will be critical for the success of this promising field. PMID- 12549771 TI - Chemoprevention of lung cancer: current status and future prospects. AB - Lung cancer is the leading cause of cancer death in the United States. The current mainstays of lung cancer therapy are surgery, radiation and chemotherapy. These interventions have produced slight declines in mortality rates in the last 5 years however, it appears unlikely that marked improvements will occur in the near future. This grim overview argues strongly for new, emerging approaches for controlling this disease. Chemoprevention is the use of specific natural or synthetic substances with the objective of reversing, suppressing or preventing carcinogenic progression to invasive cancer. Whether primary, secondary or tertiary settings, prevention has the highest potential to improve the dismal statistics associated with this cancer. Several randomized clinical or translational chemoprevention trials have been conducted. All have so far produced either neutral or harmful primary endpoint results showing that lung cancer was not prevented by alpha-tocopheral, beta-carotene, retinal, retinyl palmitate, N-acetylcysteine or isotretinoin in smokers. Secondary results supporting treatment with isotretinoin in 'never' and former smokers and data from prevention trials involving selenium and vitamin E however, are encouraging and offer a promising direction for future clinical study. Other areas of promise for future lung cancer chemoprevention study include the study of molecular markers of risk and drug activity, molecular targeting study, improved imaging techniques and new drug delivery systems. PMID- 12549772 TI - Chemoprevention of skin cancer: current status and future prospects. AB - Chemoprevention represents a relatively new and promising strategy whereby the use of natural or synthetic agents the process of carcinogenesis can be slowed, reversed or completely halted. Especially for the skin cancer, chemoprevention could be an important armamentarium because of increasing incidence of such cancers and because skin is continuously exposed to various environmental carcinogens that include both chemical agents and solar ultraviolet radiations. A wide range of compounds, both synthetic and naturally occurring has been shown to possess cancer chemopreventive effects in murine skin carcinogenesis models. Only a limited number of these agents have been tested for their efficacy in the human population. Here, we provide a brief review on the skin cancer chemopreventive potential and mechanism of action of various synthetic and natural agents. Many of these agents are present in daily diet and are supplemented or topically applied against prevention of various stages of skin cancer. We will also discuss the current status and future prospects of these agents for development as promising chemopreventive agents against skin cancer. PMID- 12549773 TI - Pathology of prostate cancer. PMID- 12549774 TI - Risk factors of relapse within eight weeks after an acute asthma exacerbation in Thai children. AB - BACKGROUND: Acute asthma relapse following treatment and discharge from hospital remains a substantial problem. Various potential risk factors for relapse have been reported including age, sex, frequency of hospitalization and emergency medications. All these factors, however, may not be generalized for all patients because of difference in prevalence, trigger factors, types of allergens, medical accessibility and psychosocial problems. OBJECTIVE: To identify factors associated with relapse following treatment for acute asthma within the next 8 weeks in Thai children. MATERIAL AND METHOD: The authors prospectively followed 91 children discharged from Ramathibodi Hospital after treatment of an asthma attack from June 1999 to December 2000. Parents were surveyed concerning their child's medical history, trigger factors, psychosocial and economic variables. Data on severity of the attack, asthma scores, emergency treatment, and response to treatment were recorded. Investigations included eosinophil count, total IgE, serum eosinophil cationic protein (ECP), skin test, methacholine bronchial challenge test, and IQ test were performed and recorded. RESULTS: Within the first week, only 6.6 per cent had relapsed and increased to 29.7 per cent by 8 weeks. Patients who suffered relapse were more likely associated with age at asthma diagnosis (OR, 2.90; 95% CI, 1.1-7.5) and 6 years of age or under (OR 4.49, CI 1.22-16.54). From the investigation results including eosinophil count, total IgE, serum ECP, skin test, methacholine bronchial challenge test, and IQ test, there was no significant difference in the factors between patients who suffered relapse and those who did not. From the psychosocial evaluation, 18 out 39 (46.2%) studied cases had significant psychosocial disorders. They were 4 cases with delayed development and mental retardation, 9 cases with parent-child relation problems, and 2 cases with serious intrafamilial disorders. Most of these patients were non-relapse cases. However, the relationship between asthma relapse and psychosocial disorders could not be ascertained since psychosocial evaluation was only performed in one-third of the study population. CONCLUSION: Among patients following acute asthma therapy, 29.7 per cent will have a relapse. The authors identified the age at onset of asthma before the age of 6 years as an important risk factor. This may help to decrease the relapse rate by more intensive and comprehensive management among patients at high risk. PMID- 12549775 TI - Risk factors of high lead level in Bangkok children. AB - Lead poisoning is the most important pollution in children which affects the developing brain and IQ. Previous studies of blood lead levels in Thai children were all higher than 10 microg/dl. OBJECTIVE: To identify risk factors and impacts associated with high lead levels (>10 microg/dl) in Bangkok children MATERIAL AND METHOD: The longitudinal study (n = 84) followed blood lead levels of a birth cohort, 6, 12, 18, 24 and 72 months of age from 1993 to 1999. Multiple cross-sectional studies (1992-1996) comprising children under 15 years of age attending the outpatient clinic, Ramathibodi Hospital (n = 511), kindergartens (n = 60), 6 primary school students (n = 564) and secondary school students (n = 377) in Bangkok, were recruited. Blood lead levels were assessed by atomic absorption spectrometry. Questionnaires to identify risk factors were completed by parents of under 2-year-old children and primary school students. IQ was assessed in the longitudinal group at 2 years of age and in primary students. RESULTS: The mean blood lead levels (microg/dl) were 5.57 +/- 2.31 at birth, 5.03 +/- 2.21 at 6 years of age in a cohort study, 6.74 +/- 2.02 in kindergartens, 9.26 +/- 3.68 in primary students and 9.03 +/- 3.65 in secondary school students. The proportion of high blood lead increased with age from 1 per cent at birth to 35 per cent in secondary school students. In the kindergartens and secondary school, males had higher lead levels than females. In the primary school study, the significant risk factors (Odd Ratio-OR) of high lead level were; living in Bangkok (6.18), male (1.67), maternal labour workers (1.79), family income lower than 3,000 baht/month (2.24), a crowded family with more than 9 members (2.22), household members whose present occupation was related to printing (4.55) or lead smelting previously (4.85). Children in the high lead group had lower weight (p = 0.0000) and height (p = 0.0000) and were slow learners determined by their teachers (p = 0.0332). CONCLUSION: The blood lead levels in Bangkok children were not high and have tended to decrease following the reduction of air lead levels because of unleaded gasoline usage. A periodic surveillance survey is still necessary to monitor blood lead level in Bangkok children especially among those with risk factors. PMID- 12549776 TI - Outcome of intracranial hemorrhage in infants with congenital factor VII deficiency. AB - The outcome of 8 episodes of intracranial hemorrhage in 7 patients (4 males, 3 females) with congenital factor VII deficiency was evaluated. Their levels of factor VII clotting activity (FVII:C) were less than 1 per cent (n = 3) and ranged from 1.7 to 2.3 per cent (n = 4). The onset varied from the first week (n = 2), first month (n = 3), and at the ages of 6, 11 and 12 months (n = 3). The replacement therapy of 10 ml/kg of fresh frozen plasma (FFP) every 6-12 hours for 5-7 days was given to 6 patients. Only one craniotomy for the removal of hematoma was performed. The seventh patient experienced two episodes of bleeding. First, she received 20 microg/kg of recombinant factor VIIa (rFVIIa) every 6 hours for 4 days (1,200 microg) followed by FFP in one episode. Second, a craniotomy for the removal of a 7 cm diameter hematoma was performed by giving 20 microg/kg of rFVIIa every 6 hours for 12 days (9,600 microg) followed by FFP in another episode. As a result of these treatments, 2 died and 5 survived with sequelae, except for one who received rFVIIa. The sequelae included seizure disorder (n = 1) and hydrocephalus (n = 3). Subsequently, the surviving patients received 15 ml/kg of lyophilized fresh plasma every 3-5 days as prophylactic treatment. In conclusion, rFVIIa in the dose of 20 microg/kg every 6 hours has been shown to be effective in controlling intracranial hemorrhage in patients with congenital factor VII deficiency. PMID- 12549777 TI - Serum diazepam levels after oral administration in children. AB - OBJECTIVE: To determine serum levels of diazepam after oral administration in children. PATIENTS AND METHOD: Forty six children admitted with febrile seizures were orally administered with 0.25 mg/kg/dose of diazepam six hourly for four doses. Trough (prior to the next dose) and peak (at 1 hour 20 minutes after the dose) serum levels of diazepam were analyzed. The patients were observed for adverse effects of the medication. RESULTS: The peak levels after 1st, 2nd, 3rd and 4th doses were above 0.15 microg/ml which is considered the therapeutic level in 93.5, 97.8, 97.7, and 100 per cent of the patients, respectively. The trough levels prior to the 2nd, 3nd, and 4th doses were greater than 0.15 microg/ml in 75.0, 84.0, and 91.3 per cent, respectively. Neither recurrent seizure nor serious adverse effects occurred in any of the patients. CONCLUSION: Serum concentrations above the therapeutic level were achieved after orally administered diazepam at 0.25 mg/kg/dose six hourly for four doses. Oral diazepam may be used as another method in the prevention of recurrent febrile seizures. PMID- 12549778 TI - The limitation of factor IX coagulant activity determination in the diagnosis of hemophilia B carriers. AB - A preliminary study of factor IX coagulant activity (FIX:C) for determining hemophilia B carriers was conducted at the Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital in Bangkok. Twenty-eight females (8 obligate, 20 potential carriers) from 17 hemophilia B families were enrolled in the study. Additionally, 25 normal females were included. They were not pregnant and not using oral contraceptives. Then, three cut-off levels of FIX:C including 50 per cent which was the commonly used level; 57 per cent which was the mean-2 SD of normal females and 75 per cent which was the level reported by Knobe and Ljung in 1999 were used for the diagnosis of hemophilia B carriers. The sensitivities of these three cut-off levels were 12.5 per cent (1/8) for 50 per cent, 37.5 per cent (3/8) for 57 per cent and 50 per cent (4/8) for 75 per cent. Also, the specificities were 100 per cent (25/25) for both 50 and 57 per cent, and 96 per cent (24/25) for 75 per cent. Although the low cut-off levels of 50 per cent and 57 per cent had low sensitivities, they yielded a high specificity (100%) compared to the higher level of 75 per cent. In the present study, the sensitivity of the cut-off level at 75 per cent was much lower than that of the study by Knobe and Ljung (93%) since the presented sample size of obligate carriers was rather small. So, enrollment of more subjects should be further carried out. In conclusion, FIX:C determination alone showed a limitation in the diagnosis of hemophilia B carriers. The addition of genetic analysis of linkage analysis or mutation detection is required for a definite diagnosis. PMID- 12549779 TI - The association between television viewing and childhood obesity: a national survey in Thailand. AB - OBJECTIVE: To determine the relationship between television viewing and obesity among Thai children. DESIGN: A one-stage cross-sectional national survey. SETTING: National communities in Thailand. SUBJECTS: Four thousand one hundred and ninety-seven children aged 6 to 12 years old. RESULTS: By using the new national growth reference as a normative data, 262 (6.2%) children in this survey were obese. According to the caregivers, 62 per cent of children watched television I to 3 hours per day. Children who spent more than 3 hours per day watching television were more likely to be obese than the children who watched television less with the odd ratio of 1.8 (95% confidence interval = 1.2-2.8). CONCLUSION: With a one-stage design for determining the association between television viewing and childhood obesity, Thai school age children who watched television more than 3 hours per day were more likely to be obese with statistical significance. PMID- 12549780 TI - Iron status of one-year-old infants in a well baby clinic. AB - Seventy-two healthy infants (37 males, 35 females) attending a private well baby clinic were enrolled in the study. Their mean birthweights and body weights at one year of age were 3,079 grams and 10 kilograms, respectively. Blood samples were drawn approximately on their first birthday for evaluating the iron status. Complete blood count, hemoglobin (Hb) typing and DNA analysis for common carrier status of thalassemia and hemoglobinopathis were also determined. According to the infants of serum ferritin, the patients were classified into 4 groups: group 1, iron deficiency anemia (Hb <11 g/dl and ferritin <12 ng/L) in 1 infants (1.4%); group 2, iron deficiency without anemia (Hb >11 g/dl and ferritin <12 ng/L) in 5 infants (6.9%); group 3, borderline iron depletion (ferritin 12-30 ng/L) in 39 infants (54.2%); group 4, iron sufficiency (ferritin >30 ng/L) in 27 infants (37.5%). The iron deficiency state emerged as 8.3 per cent (6/72). There was no significant difference of levels of Hb and mean corpuscular volume (MCV) among the infants with iron deficiency without anemia, borderline iron depletion and iron sufficiency. The results also revealed that 25 out of 72 (34.7%) infants were carriers of thalassemia and hemoglobinopathies. The carrier infants had significant lower Hb and MCV than those of the non-carrier infants with the p values of 0.004 and 0.000, respectively; while their serum ferritin levels were not significantly different. Additionally, the association of carrier and iron deficiency state was further evaluated. The Hb and MCV among carrier infants with and without iron deficiency were not significantly different. Six infants with carrier state were found to have slightly decreased levels of Hb ranging from 10.3 to 10.9 g/dl with the ferritin ranging from 18.7 to 382.9 ng/L while the remainders had Hb of >11 g/dl. Therefore, 7 out of 72 (9.2%) infants had anemia (Hb <11 g/dl) which was caused by the carrier state of thalassemia and hemoglobinopathies (n=6) and iron deficiency anemia (n=1). The risk factors of iron deficiency status were associated with feeding regimen including continuation of breast feeding until one year of age without adequate haem iron supplement, exclusive formula feeding, inadequacy of solid food supplement with only one meal per day and excluding haem iron from animal liver without substitution. The infants with risk factors had significantly lower levels of serum ferritin (mean 14.1 +/- 1.7 ng/L) than those without risk factors (mean 31.9 +/- 1.9 ng/L) with a p-value of 0.000. In conclusion, adequate haem iron supplement in 3 meals of solid food is essential for the prevention of iron deficiency status in one-year-old infants. PMID- 12549781 TI - Effects of sodium butyrate and acidic fibroblast growth factor on TDC32300 cultured cells. AB - BACKGROUND: Many studies have demonstrated that transition duct cells (TDC) are facultative liver stem cells. Our laboratory established TDC32300 cell lines with hepatic progenitor markers. The authors proposed that cell culture using sodium butyrate (NaBut) and acidic fibroblast growth factors (aFGF) may support the differentiation of TDC32300 cells along the hepatic lineage. METHODS: TDC32300 cells were cultured in four different conditions 1) STON media alone; 2) STON with NaBut in 3 different concentrations, 1 mM, 3.75 mM and 5 mM; 3) STON with aFGF; and 4) STON with aFGF and dexamethasone. After day 5, the cultured cells were fixed and stained with monoclonal antibodies to rat liver antigens and anti proliferating nuclear antigen (PCNA). RESULTS: Proliferation of TDC32300 cells cultured in the high concentration of NaBut (3.75 and 5 mM) was inhibited. This phenomenon was confirmed by the reduction in cell number and decrease in PCNA expression. Irrespective of the concentration, NaBut did not alter the phenotype of the TDC32300 cultured cells. aFGF with or without dexamethasone also did not alter the phenotypic characteristic of TDC32300 cells. CONCLUSION: TDC32300 cells may not be the hepatic progenitors or that their differentiation may require other culture conditions. PMID- 12549782 TI - Prevalence and clinical characteristics of dyslexia in primary school students. AB - BACKGROUND: Dyslexia is the most common subtype of learning disabilities with a prevalence ranging from 5-10 per cent. The central difficulty in dyslexia is the phonological awareness deficit. The authors have developed a screening test to assess the reading ability of Thai primary school students. OBJECTIVE: 1. To study the prevalence of dyslexia in first to sixth grade students at Wat Samiannaree School. 2. To study the clinical characteristics such as sex, neurological signs, verbal intelligence and comorbid attention deficit hyperactive disorder (ADHD) of the dyslexia group. METHOD: A total of 486 first to sixth grade students were administered "Raven's progressive matrices test" for estimation of intellectual functioning. Those who scored below the fifth percentile were labeled as mental retardation and excluded from the study. The students' reading ability was evaluated by 3 steps; first by classroom teachers using some items of the screening test, second by the researchers examining some more items individually, and third by the special educator assessing more details in reading and phonology. The students who had a reading ability two-grade levels below their actual grades and impairment in phonology were diagnosed with dyslexia. RESULTS: The prevalence of dyslexia and probable dyslexia were found to be 6.3 per cent and 12.6 per cent, respectively. The male to female ratio of dyslexia was 3.4:1. The dyslexia group had significantly lower Thai language scores than those of the normal group (p < 0.05). All of the dyslexia group had a normal grossly neurological examination but 90 per cent showed positive soft neurological signs. Mean verbal intellectual quotient score in the dyslexia group assessed by using Wechsler Intelligence Scales for Children--Revised was 76 +/- 7. The comorbid ADHD was 8.7 per cent in the dyslexia group. CONCLUSION: Dyslexia was a common problem among primary school students in this study. Further studies in a larger population and different socioeconomic statuses are required to determine the prevalence of dyslexia in the general population. The authors suggest evaluating the reading ability carefully by using a test that can detect phonological awareness deficit in all children who have learning problems. PMID- 12549783 TI - Accuracy of glucose meters in measuring low blood glucose levels. AB - BACKGROUND: Hypoglycemia is an emergency condition requiring treatment as soon as possible. Therefore, rapid and reliable blood glucose measurements are necessary. There are 2 systems of glucose meters (GMs), the reflectance photometer system (RPS) and the electrochemical biosensor system (BSS). GMs are widely used in monitoring blood glucose (BG) in patients with diabetes. BG values measured by GMs have been confirmed to be accurate especially in measuring normal and high BG levels. However, the data on the accuracy of GMs in measuring low BG levels are limited. OBJECTIVE: To compare accuracy and reliability of different systems of GMs in the measurement of low BG values. PATIENTS AND METHOD: Venous and capillary whole blood specimens were collected from patients who were investigated for pituitary dysfunction. The patients underwent an insulin-induced hypoglycemia test by intravenously administering human regular insulin. The low BG level was defined as having venous plasma glucose (PG) of less than 60 mg/dl (mean +/- SD = 36.59 +/- 9.19, n = 54). Capillary blood samples were obtained from fingertips. Venous BG (vBG) and capillary BG (cBG) were measured by GMs. Venous PG which considered a reference value was measured by the glucose dehydrogenase method. RESULTS: The correlation coefficient (r) between vBG measured by GMs-RPS and PG was 0.86 (p < 0.001), whereas, that between vBG by GMs BSS and PG was 0.75 (p < 0.001). Similarly, the r between cBG by GMs-RPS and PG was 0.73 (p < 0.001), whereas, that between cBG by GMs-BSS and PG was 0.69 (p < 0.001). The mean difference between vBG by GMs-RPS and PG values was 0.01 +/- 4.90 mg/dl, whereas, that between vBG by GMs-BSS and PG values was 10.50 +/- 7.07 mg/dl which was significantly greater than the former (p<0.001). Moreover, the mean difference between cBG by GMs-RPS and PG values was 14.45 +/- 8.76 mg/dl, whereas, that between cBG by GMs-BSS and PG values was 23.87 +/- 9.48 mg/dl which was also significantly greater than the former (p < 0.001). These results demonstrated that vBG measured by GMs-RPS was comparable to PG values. In addition, cBG values by GMs-RPS and GMs-BSS were approximately 14 mg/dl or 38 per cent and 24 mg/dl or 65 per cent greater than PG values, respectively. CONCLUSION: In measuring low blood glucose levels, glucose meters using the reflectance photometer system are more accurate than those using the electrochemical biosensor system. PMID- 12549784 TI - Respiratory syncytial virus-associated lower respiratory tract infection in under 5-year-old children in a rural community of central Thailand, a population-based study. AB - The population-based cohort study on the epidemiology of respiratory syncytial virus (RSV)-associated lower respiratory tract infection (LRI) (RSV-LRI) was conducted in Takhli district from November 1998 to February 2001. The incidence of RSV-LRI was 12.6/1,000 child-year and 5.8/ 1,000 child-year during the first and second year, respectively. RSV accounted for 35.8 per cent of all LRI cases during the first year and significantly decreased to 17.5 per cent during the second year. Three-quarters of RSV-LRI occurred among children under 2 years old (76.6% during the first year and 62.2% during the second year). Most of RSV-LRI in both years occurred from July to October. Risk factor for morbidity of RSV infections were age less than or equal to 2 years (OR = 2.38, 95% CI = 1.22-4.67 p = 0.009) and sleeping with more than 3 persons in the patient's bedroom (OR = 2.92,95% CI = 1.42-6.00, p = 0.002). Most RSV-LRI (63.9%) were clinically diagnosed as having pneumonia. No RSV-LRI deaths were detected. During the first year, RSV subtype B was predominate, in contrast to the second year when subtype A was more predominate. Further research to determine the annual change in subtype of RSV-LRI and correlation of severity of disease with specific subtypes needs to conducted in order to prepare for the future introduction of a vaccine. PMID- 12549785 TI - Treatment of Kawasaki disease with a moderate dose (1 g/kg) of intravenous immunoglobulin. AB - BACKGROUND: Kawasaki disease is an acute febrile illness recognized most often in young children. Coronary abnormality is the most serious complication preventable with intravenous immunoglobulin (IVIG) administration. Various treatment regimens of IVIG have been reported. OBJECTIVE: To determine initial treatment failure and prevalence of coronary artery abnormality (CAA) in Kawasaki disease (KD) treated with a moderate dose (1 g/kg) of intravenous immunoglobulin (IVIG). METHOD: All patients with a diagnosis of KD who had initial treatment with 1 g/kg of IVIG at Ramathibodi Hospital between 1994 and 1998 were reviewed retrospectively. RESULTS: Thirty-one of 41(76%) patients responded completely to a single treatment with a moderate dose of IVIG (group A). The second dose of 1 g/kg of IVIG was required in 7 patients (17%) due to persistent fever more than 48 hours after the initial treatment (group B), and 3 patients (7%) required 3 doses of 1 g/kg of IVIG due to persistent fever after the second dose (group C). During the convalescent phase, there were 19 per cent, 29 per cent and 100 per cent of the patients in group A, B and C, respectively who developed CAA with an overall rate of 27 per cent. After 1-year follow-up, the prevalence of CAA had decreased to 3 per cent, 0 per cent and 67 per cent in the according groups with overall rate of 9.6 per cent. Only 1 patient in group C developed a giant aneurysm of the right coronary artery. CONCLUSION: The efficacy of a moderate dose (1 g/kg) of IVIG in preventing CAA is lower than that of the high dose regimen (2 g/kg) reported previously. Short duration of fever before starting IVIG and low hemoglobin level may be the risk factors of unresponsiveness to moderate-dose IVIG. PMID- 12549786 TI - Earlier onset of pubertal maturation in Thai girls. AB - BACKGROUND: Recent studies in the USA and Hong Kong demonstrated the onset of puberty in girls has shifted toward a younger age. Based upon previous studies on variations of pubertal maturation in Thai girls, the secular trend has also moved toward an earlier age. The latest study in 1995 revealed the mean age of menarche was 12.3 years. OBJECTIVES: To identify the onset of puberty, menarche and pubarche in female children and adolescents in 2 Bangkok schools. METHOD: Three hundred school girls aged 9-19 years were enrolled in the study. Data were collected from January 1997 through December 1999. Assessment of pubertal staging by Tanner's criteria was performed by a trained pediatrician. All were in good physical health and had normal height and weight. The median ages of thelarche, menarche and pubarche were estimated by probit analysis. All other parameters were expressed as mean +/- SD. RESULTS: The median ages of thelarche and pubarche were 9.4 and 11.1 years, respectively. Two hundred and twenty one girls had experienced menstruation. The median age of menarche was 11.2 years, whereas, the mean age was 12.1 years. Most girls reached near final adult height after 14 years old. CONCLUSION: The secular trend in decline of the ages of thelarche (or puberty) and menarche was observed in Bangkok girls. Further study in a larger population including a younger age group is required to define the current reference interval of onset of puberty. PMID- 12549787 TI - Neurobiologic risk score and long-term developmental outcomes of premature infants, birth weight less than 1,250 grams. AB - OBJECTIVE: To determine whether neurobiologic risk score (NBRS) would continue to correlate with developmental outcomes. METHOD: An observational cohort consisting of 258 surviving infants who returned to the follow-up clinic with a mean age 22 months' corrected age. Both univariate and multivariate analysis were performed to identify risk factors and to assess the predictive value of NBRS. RESULTS: Forty-eight to 53 per cent of these infants had growth parameters < 25th percentile for age. Seventeen and 18 per cent respectively had mental developmental index (MDI) and psychomotor developmental index (PDI) on the Bayley Scales less than 70 and 14 per cent developed cerebral palsy (CP). NBRS demonstrated a significant correlation with the outcome (p < 0.001). In infants with NBRS > or = 8, 48 per cent had MDI < 70 and 68 per cent, had PDI < 70. At a similar NBRS cutoff, specificity and negative predictive value (NPV) were 86 and 96 per cent, respectively. Logistic regression indicated that birth weight and gestational age were the most significant, independent variables for predicting poor outcomes. CONCLUSION: Very preterm infants in the present study were at risk for abnormal developmental outcomes. NBRS demonstrated a very high specificity and NPV and may be a useful index to identify those who need early intervention. PMID- 12549788 TI - The optimal dose of potassium citrate in the treatment of children with distal renal tubular acidosis. AB - BACKGROUND: Distal renal tubular acidosis (RTA) is a common cause of intractable calcium nephrolithiasis. In adults, the use of potassium citrate (PC) in distal RTA effectively decreases metabolic acidosis and the risk of calcium oxalate stone but it cannot decrease the risk of calcium phosphate stone. However, there is no report for the optimal dose of PC and the risk of calcium stone in distal RTA in children. OBJECTIVE: To evaluate the optimal dose of PC that minimizes the risk of calcium nephrolithiasis in children with distal RTA. METHOD: Prospective study PATIENTS: Children who have distal RTA and were followed-up for 4 months. Patients were studied in a control phase, 1 month of PC 2 mEq/kg/day, 2 months of PC 3 mEq/kg/day and 1 month of PC 4 mEq/kg/day. The urine specimens of 41 normal children were measured for the reference value of the parameters determining the risk of calcium stone. RESULTS: Eight children (mean age of 10 +/- 3.7 years, female : male = 6: 2) with distal RTA were studied during the control phase and after receiving PC 2 mEq/kg/day for I month. Treatment with PC 2 mEq/kg/day was not able to normalize serum bicarbonate and caused no significant change in the urine citrate/creatinine ratio, and activity production of calcium phosphate stone but it caused a significant decrease in the urine calcium/citrate ratio. Although PC 3 mEq/kg/day for I month normalized plasma bicarbonate, only this dose given for 2 months caused a significant increase in the urine citrate/creatinine ratio and urine calcium/ citrate ratio to values that were not different from normal children, while the activity production of calcium phosphate stone did not decrease to normal level. The effect of PC 4 mEq/kg/day was similar to that of 3 mEq/kg/day. CONCLUSION: Potassium citrate 3 mEq/kg/day for 2 months effectively normalized serum bicarbonate and decreased the risk of calcium oxalate stone but this treatment was theoretically unable to reduce the risk of calcium phosphate stone in children with distal RTA. PMID- 12549789 TI - Predictive values of serial C-reactive protein in neonatal sepsis. AB - BACKGROUND: Infection is one of the major problems in neonates. The diagnosis of neonatal septicemia is difficult to establish based on the clinical criteria alone. However, empirical treatment should not be delayed because of the high mortality. Laboratory tests used to support diagnosis have shown variable predictive values. C-reactive protein (CRP), an acute phase protein, increases in inflammatory disorders and tissue injury. Serial CRP have been shown to be more useful than a single measured CRP in the diagnostic evaluation of neonates with suspected infection. OBJECTIVES: 1. To evaluate the diagnostic accuracy of serial CRP in neonatal sepsis. 2. To compare the diagnostic values between CRP and leukocyte index from a complete blood count (CBC). METHOD: A prospective observational study included newborn infants, aged > 3 days and diagnosed with clinical sepsis, who were admitted in the newborn intensive care unit and special care nursery at Ramathibodi Hospital during a 14-months period. Newborn infants who received antibiotics prior to septic work up were excluded. CRP levels were measured initially at the time of septic work-up and at 24-48 hours later. Investigations for infection included CBC, blood culture and urine culture. Radiological study and lumbar puncture were performed if clinically indicated. Based on clinical and biological data, diagnosis of infants can be categorized into 4 groups as follows; (1) proven sepsis with positive culture, (2) localized infection with negative culture, (3) probable infection (clinically consistent with sepsis, negative culture without localized infection), and (4) no infection (findings not consistent with sepsis and antibiotics were discontinued within 3 days). Diagnosis was made before the CRP results were known. RESULTS: Of 76 newborn infants with 90 episodes of clinical sepsis, there were 24 episodes of proven sepsis, 11 episodes of localized infection with negative culture, 18 episodes of probable infection and 37 episodes of no infection. Serial CRP had better predictive values than those of CBC. The sensitivity, specificity, positive predictive value, and negative predictive value of CRP for proven sepsis and localized infection at cutoff point > or = 5 mg/L were 100 per cent, 94 per cent, 91.6 per cent and 100 per cent respectively. False positive CRP were found in post-operative patent ductus arteriosus ligation, intracerebral hemorrhage, and post resuscitation with chest compression. To improve the predictive value of CBC, analysis of the receiver operating characteristic (ROC) curve showed that the predictive value of CBC for sepsis would be enhanced by using abnormal leukocyte index 2 2 parameters. CONCLUSIONS: Predictive value of CRP could be enhanced by serial rather than a single measurement. Serial CRP showed very high predictive values for diagnosis of neonatal sepsis and were better than those of leukocyte indices of CBC. PMID- 12549790 TI - Comparison of the effectiveness between the adapted-double phototherapy versus conventional-single phototherapy. AB - BACKGROUND: Jaundice is very common in neonates during the first few days of life. Severe hyperbilirubinemia is potentially neurotoxic, resulting in bilirubin encephalopathy. Phototherapy has been used widespread and proven to be effective in lowering serum bilirubin. The efficacy of phototherapy depends on the type of light-source, the intensity of light and the area of skin exposed. Double phototherapy with a fiber-optic blanket has been reported to be more effective in reducing bilirubin than conventional phototherapy. However, a fiber-optic blanket or bili-bed is very expensive, thus the authors designed the adapted-double phototherapy by using ordinary fluorescent lamps that are much cheaper and more easily available. OBJECTIVE: To compare the efficacy and safety of adapted-double phototherapy (DP) using daylight fluorescent lamps to conventional phototherapy (CP) in healthy term infants with hyperbilirubinemia. DESIGN: Prospective randomized controlled trial study. METHOD: Term infants who met phototherapy criteria were randomized into two groups of phototherapy; the adapted DP placing daylight fluorescent lamps 38 cm above and 32 cm below the crib to produce 9-10 microw/cm2/nm (n = 24) versus CP of similar irradiance (n = 27). RESULT: There were no significant differences between the two groups with respect to birth weight, gestational age, hematocrit at enrollment and cause of hyperbilirubinemia. Infants in the DP group had higher bilirubin levels and were slightly older at the time of enrollment than those in the CP group. The bilirubin reduction rate after therapy was significantly greater in the DP compared to CP; 0.22 +/- 0.12 mg/dl/h vs 0.14 +/- 0.1 mg/dl/h on day 1 of therapy (p = 0.02) and 0.16 +/- 0.11 mg/ dl/h vs 0.1 +/- 0.05 mg/dl/h on day 2 (p = 0.06). Duration of phototherapy was shorter in DP; 34.9 +/- 12.6 h vs 43.7 +/- 17.5 h in CP group (p = 0.039). No differences in side effects were found between two groups. CONCLUSION: The adapted-DP using daylight fluorescent lamps in this study has proven to be safe and more effective in reducing bilirubin than CP. It could be an alternative model for intensified phototherapy as it produces a reasonable cost-effectiveness and is easy to apply. PMID- 12549791 TI - Acute viral lower respiratory infections in children in a rural community in Thailand. AB - The present study was conducted as a population based cohort in a rural community of Amphoe Takhli, Nakhon Sawan province for the determination of the prevalence of acute viral lower respiratory infection (ALRI) in pediatric cases under 5 years of age from November 1998 to February 2001. There were 472 ALRI episodes during the study period; and there were 5 cases who contracted ALRI twice. The etiologic agents were determined by indirect immunofluorescence (IIF) test using specific monoclonal antibodies for the staining of exfoliated cells in nasopharyngeal aspirate (NPA) samples. The slides of fixed cells were prepared by Takhli Hospital and posted in ambient temperature to the Virology Laboratory, Siriraj Hospital where they were stained and examined. Among 472 episodes of ALRI, 170 (36.0%) viral agents were found. Viral agents were associated with 41.4 per cent of all pneumonic cases. Respiratory syncytial virus (RSV) was the most common virus observed in the present study; and it was also the most common virus associated with pneumonia and bronchitis. RSV subgrouping was performed directly in NPA samples by IIF test using a panel of subgroup specific monoclonal antibodies. RSV subgroup B predominated over subgroup A in the first study year, and it was vice versa in the second year. Overall, more cases of subgroup B were found which was in contrast to what the authors had reported in the previous study. Prevalence of RSV was seasonal dependent, the epidemic was seen during the rainy season with peaks in August or September of each year. As the method of viral identification was limited to IIF only, therefore, fewer viruses were detected. Parainfluenza viruses were detected as the second most common viral agent, the viruses spread during early summer with peaks in February or March of each year. However, its association with croup could not be demonstrated which may be due to the insensitivity of IIF in the diagnosis of non-RSV infection. Nevertheless, in terms of feasibility to investigate the disease in a rural area, IIF is economic, convenient and rapid; and gives enough information for the nationwide plan of a health care development system. PMID- 12549792 TI - Oral erythromycin for treatment of feeding intolerance in preterm infants: a preliminary report. AB - Feeding intolerance is a common problem in preterm infants resulting in a prolonged hyperalimentation which is associated with an increased risk of serious and sometimes even life threatening complications, including cholestasis jaundice, liver impairment, nutritional deficiency, biochemical rickets and catheter-related septicaemia. Erythromycin, a commonly used macrolide antibiotic, has been reported as having potent prokinetic properties and enhancing gastrointestinal motor activity. The authors, therefore, conducted a preliminary study of oral erythromycin for the treatment of feeding intolerance in preterm infants to evaluate the safety and efficacy of this drug. AIM: To evaluate the safety and efficacy of oral erythromycin as a prokinetic agent in promoting enteral feeding in preterm infants with feeding intolerance. METHOD: Preterm infants, gestational age (GA) < or = 36 wk, who met the feeding intolerance criteria, were enrolled in the study. Inclusion criteria included infants who received enteral feeding less than half of full feeding or less than 75 ml/kg/day by day 14 post-natal age or gastric residual > or = 50 per cent of a given amount of feeding, more than 2 consecutive feeds by day 7 post-natal age. All patients received oral erythromycin ethylsuccinate 12 mg/kg every six hours for 2 days, then 3 mg/kg every six hours for 5 days. The times taken to establish full enteral feeding after the drug treatment and time to stop hyperalimentation were recorded. Potential adverse effects of erythromycin were assessed. Response to treatment was defined as decreased gastric residual < 30 per cent of a similar amount of the previous feed and was able to continue to full feeding. RESULTS: Ten preterm infants were enrolled in this study with a mean GA of 30.8 weeks (26 35), mean birth weight of 1,489 g (range 900-2,560 g) and mean age at entry of 9.2 days (range 7-12 days). Nine of 10 infants responded to treatment within 24 hours. The average time to establish full enteral feeding after the drug treatment was 6.6 days (range 4-10 days). None of the infants developed adverse effects such as vomiting, diarrhea, or pyloric stenosis. CONCLUSION: The preliminary data indicates that oral erythromycin is effective and safe in facilitating enteral feeding in preterm infants with feeding intolerance. Infants can achieve full feeding within a week after treatment, and this may shorten the course of hyperalimentaiton. Further randomized controlled trials are warranted. PMID- 12549793 TI - Modified percutaneous endoscopic gastrostomy tubes: experience in Thai children. AB - BACKGROUND: Percutaneous endoscopic gastrostomy (PEG) is a well-established alternative to open gastrostomy for providing long-term enteral nutrition. Although the commercial PEG tube is available and suitable for the procedure, its cost is relatively high for low socioeconomic people. Therefore, modified PEG tubes have been used in our hospital. OBJECTIVES: To evaluate the outcome and complications of PEG performed in children at Ramathibodi Hospital and compare the results between the commercial PEG and modified PEG tubes. METHOD: All children who had PEG performed at Ramathibodi Hospital, from January 1999 to May 2002, were included in the study. The demographic data, indications for PEG, types of PEG tube, outcomes and complications were retrospectively reviewed. The modified PEG tube was made by connecting a Malecot four-wing catheter to the previously used, re-sterilized distal part of a commercial PEG tube. RESULTS: PEG was performed on 34 children, aged 4 months to 13 years, and successfully placed in 30 children (88.2%). The commercial and modified PEG tubes were used in 20 cases and 10 cases, respectively. Early complications occurring in the first 7 days post-procedure were found in 9 cases (30%) as follow: peritonitis (1 case), peristomal wound infection (7 cases), and subcutaneous emphysema (1 case). Late complications occurring at more than 7 days post-procedure were found in 15 cases (50%) and all were minor problems. There was no difference in complication rates between the 2 types of PEG tubes. CONCLUSION: PEG is safe even in small infants. Minor complications are common but can be simply managed. The modified PEG tube is an alternative for a commercial PEG tube in an unaffordable situation. PMID- 12549794 TI - Nutritional support in pediatric patients undergoing bone marrow transplantation. AB - BACKGROUND: Children undergoing bone marrow transplantation (BMT) are prone to develop severe gastrointestinal (GI) complications and metabolic imbalance which consequently impair their nutritional status. Nutritional support is an important adjunctive treatment during BMT. OBJECTIVE: To assess GI complications, metabolic complications and nutritional outcome of children undergoing BMT with nutritional support intervention. METHOD: Retrospective study of 20 children (median age 6.8 years, 11 males) undergoing BMT at Ramathibodi Hospital from March 1995 to July 2000 was conducted. Their medical records were reviewed. RESULTS: The patients underwent autologous (n = 9) and allogenic BMT (n = 11). Median z-scores of weight for age, height for age and weight for height were 0.06 +/- 1.93, -0.55 +/ 1.18 and 0.48 +/- 1.94, respectively. Nineteen patients had vomiting for 9.8 +/- 5.5 days. Eighteen patients developed diarrhea for 9.6 +/- 7.2 days. The durations of vomiting and diarrhea, as a percentage of total hospital days, were 33.5 +/- 16.3 per cent and 30.4 +/- 17.0 per cent, respectively. There were no differences between the patients with autologous and allogenic BMT regarding these durations. All patients needed enteral and/or parenteral nutrition support for 21.0 +/- 7.7 days except for one patient who could take adequate oral intake. The duration of enteral nutrition support was not significantly different between the groups but the duration of parenteral nutrition support was significantly longer in the allogenic group. Metabolic complications were hypokalemia, hypophosphatemia and one case of arrhythmia secondary to hypomagnesemia. All patients developed febrile neutropenia but none developed catheter-related sepsis. The length of hospital stay was 30.5 +/- 10.2 days. The median z-score of weight for height on the day of discharge was 1.08 +/- 2.03. CONCLUSION: Children undergoing BMT usually have GI symptoms of vomiting, diarrhea and mucositis as well as metabolic imbalances such as hypokalemia, hypophosphatemia and hypomagnesemia. Despite these complications, their nutritional status could be restored by proper nutritional support. PMID- 12549795 TI - Effect of oral vitamin E supplementation in children with cholestasis. AB - OBJECTIVE: Malabsorption and deficiency of vitamin E are common consequences of chronic cholestasis. The objective of this study was to determine vitamin E status by using plasma vitamin E/total lipid ratio (E/L) in children with cholestasis during supplementation with 20 IU/kg/day and 100 IU/kg/day of oral vitamin E capsule, and 50 IU/kg/day of cold water soluble form (CWSIF) of vitamin E. METHOD: Children with cholestasis who were being supplemented with 20 IU/kg/day of oral vitamin E capsule (dl-alpha-tocopherol) were enrolled into this study. After initial evaluation for vitamin E status and liver function, doses of oral vitamin E supplementation were increased to 100 IU/kg/day for 1 month. Then, supplementation was switched to 50 IU/kg/day of CWS/F vitamin E for 1 month. Vitamin E status was assessed by using plasma E/L after each period of supplementation. RESULTS: Eleven children with biliary atresia, aged between 2 and 18 months, were studied. Their median weight standard deviation score (SDS) was -1.35 and median height SDS was -1.26. The medians of serum direct bilirubin and total bilirubin were 6.5 and 12.9 mg/dl, respectively. Only 2 and 3 out of 9 children had plasma E/L above normal cut-off levels during supplementation with 20 and 100 IU/kg/day of vitamin E capsule, respectively. Additionally, 4 of 9 children had plasma E/L above normal cut-off level after one month's supplementation with 50 IU/kg/day of CWS/F vitamin E. All the responders had serum bilirubin levels less than 4 mg/dl while the remainder with serum direct bilirubin level more than 4 mg/dl had their plasma E/L below normal cut-off levels in spite of any vitamin E supplementation. CONCLUSION: Oral vitamin E supplementation with 20 IU/kg/day and 100 IU/kg/day of vitamin E capsule and with 50 IU/kg/day of CWS/F vitamin E were able to normalize vitamin E status in a few cholestatic children who had serum direct bilirubin levels less them 4 mg/dl. In cases of serum direct bilirubin more than 4 mg/dl, neither of vitamin E supplementations was able to correct the vitamin E deficiency status. PMID- 12549796 TI - Dead space ventilation in volume controlled versus pressure controlled mode of mechanical ventilation. AB - Dead space ventilation (VD) is one of the important measurements that indicates the ventilatory efficiency of a patient who requires mechanical ventilation. However, VD is not constant and can change according to the pathology in the lungs, ventilatory patterns, perfusion and ventilation-perfusion matching. The objective of this study was to measure and compare the dead space in pediatric patients who were using pressure controlled and volume controlled modes of mechanical ventilatory by measuring the difference between arterial PCO2 and end tidal PCO2 [P(a-ET)CO2]. From November 1996 to March 1997, 12 patients who were admitted to the pediatric intensive care unit and needed ventilator support for various reasons, were enrolled in the study. Their ages ranged from 2 to 15 years. The mechanical ventilator (Benett 7200 or Servo 900C) setting during VD measurement i.e. tidal volume, inspiratory time and positive end expiratory pressure were kept constant between changing from pressure controlled to volume controlled mode or vice versa for twenty minutes in order to allow adequate time for equilibration. The P(a-ET)CO2 between volume controlled and pressure controlled mode were 3.1 and 2.6 torr (p = 0.5) and peak inspiratory pressure were 20.0 and 17.3 torr (p = 0.01), respectively; whereas mean airway pressure, PaO2, O2 saturation and heart rate revealed no significant difference between these two modes. The authors concluded that VD in pressure controlled mode from the present study was not significantly different from VD when using volume controlled mode of mechanical ventilation in the same patient. However, VD will change according to the pathophysiologic change in respiratory system and can be used for monitoring of ventilatory pattern of patients in the pediatric intensive care unit. PMID- 12549797 TI - Henoch-Schonlein purpura: clinical manifestations and long-term outcomes in Thai children. AB - The clinical features of 47 children with Henoch-Schonlein purpura (HSP) are presented. The most common ages at presentation ranged from 3-5 years. Duration of data collection was 60 months. The peak incidence was from December to February. The organ involvements included skin (100%), gastrointestinal tract (74.5%), renal (46.8%) and joint (42.6%). Renal involvement was detected within the first 2 months in 16 cases (72.7%) but was delayed until 6 months after diagnosis in 6 cases. No risk factors for renal involvement could be identified. The mean duration of follow-up was 2.6 years (range 1-5 years). Six out of 16 (37.5%) patients had residual renal diseases but none were end stage. Recurrent episodes of abdominal pain and skin purpura were found in a few cases during the first year. Overall prognosis of HSP is good and long-term morbidity is predominantly associated with renal involvement. Patients with initially normal urinalysis should have sequential urinary examination at least for 6 months. PMID- 12549798 TI - Vitamin A status in premature infants. AB - BACKGROUND: Premature infants are at risk of vitamin A deficiency due to inadequate transplacental transport, inadequate storage and increased tissue utilization. Previous studies reported a significant decrease in serum vitamin A levels in premature infants at birth compared to those of full term infants. OBJECTIVE: To determine serial changes of plasma vitamin A status during the first month of life in 19 healthy, very low birth weight premature infants. METHOD: Subjects were fed with premature infant formula and received multivitamin supplementation. Plasma vitamin A concentrations were measured at 7, 14, and 30 days of age. RESULTS: Plasma vitamin A levels at 7,14 and 30 days of age were 24.63 +/- 6.08, 30.97 +/- 5.26 and 30.68 +/- 7.14 microg/dl, respectively. Plasma vitamin A levels at age 7 days were significantly lower than those at 14 and 30 days of life (p < 0.001). Three infants out of 19 (16%) had low plasma vitamin A (<20 microg/dl) at 7 days. At 14 and 30 days of age, all infants had normal plasma vitamin A levels. CONCLUSION: The results suggested that healthy premature infants were prone to subclinical vitamin A deficiency during the first week of life which could be treated by adequate enteral feeding and routine multivitamin supplementation. A high dose of vitamin A supplementation was not necessary in healthy premature infants. PMID- 12549799 TI - Effect of bifidobacterium Bb12 with or without Streptococcus thermophilus supplemented formula on nutritional status. AB - Acute diarrhea is a common cause of infant morbidity and mortality. Probiotic supplemented infant formula is one of the effective methods for prevention of rotavirus diarrhea. Other benefits of the probiotics supplemented formula were evaluated by monitoring the growth of the children. A double-blind, placebo controlled trial was done in 148 children aged 6-36 months. They were divided into 3 groups: the Bb12 group, 51 children received infant formula with Bifidobacteria Bb12 supplement; the Bb12+ST group, 54 children received infant formula with Bb12 and Streptococcus thermophilus supplement; and the control group, 43 children received infant formula without supplement. The mean weight Z score according to WHO reference standard of the Bbl2 group was -1.8 +/- 0.12, the Bb12+ST group was -1.4 +/- 0.11 and the control group was -1.8 +/- 0.13 at entry. The mean weight Z-score of children after 6 month showed that the children in the Bbl2+ST group had the highest increase in weight which was increased from 1.4 +/- 0.11 to -0.9 +/- 0.12 compared to the Z-score of the Bb12 group which had increased from -1.8 +/- 0.12 to -1.2 +/- 0.13 and in the control group from -1.8 +/- 0.13 to -1.7 +/- 0.25. In terms of the mean height Z-score, the Bb12 group was -2.7 +/- 0.14 to -1.7 +/- 0.16 which was higher than the Bb12+ST group (- 2.2 +/- 0.13 to -1.7 +/- 0.13) but was not different from the control group. However, the mean weight/height Z-score of the Bbl2+ST group had approached the reference standard (Bb12 group -0.1 +/- 0.11 to -0.1 +/- 0.13, Bb12+ST group -0.1 +/- 0.10 to 0.3 +/- 0.17, control group -0.4 +/- 0.12 to -0.1 +/- 0.16). Data showed that children who received the probiotics supplement formula had better growth during the 6 month period. PMID- 12549800 TI - Prevalence and clinical characteristics of attention deficit hyperactivity disorder among primary school students in Bangkok. AB - Attention deficit hyperactivity disorder (ADHD) is an important disorder because it is the most prevalent chronic health condition affecting school aged children. Children with ADHD are at risk for academic and behavior problems. There are several studies in many countries worldwide. In Thailand, there have been a few published papers about ADHD. Most of them were studies in a clinically referred population. Four hundred and thirty-three first to sixth grade students from Wat Samiennaree School were included in this study. All children were administered Raven's progressive matrices test for estimation of intellectual functioning and were observed for their behavior in the classrooms by one researcher. Their demographic data was collected by questionnaires. The revised Conners rating scales were scored for each student. Students whose parents did not score the Conners parent rating scale were excluded. The parents of students, whose scores were positive for ADHD, were interviewed according to DSM IV criteria. 353 (81.5%) students from 433 were enrolled in this study. 23 students were diagnosed with ADHD making a prevalence of 6.5 per cent. There were 11 boys and 12 girls. The ratio of male to female was 1:1.09. The ADHD students had lower scores in mathematics than the group without this diagnosis with statistical significance (p = 0.006). PMID- 12549801 TI - Economic burden in management of acute lower respiratory infection, patients' perspective: a case study of Takhli District Hospital. AB - A cross-sectional descriptive study to determine the economic burden in management of acute lower respiratory infection from the patient's perspective was conducted at Takhli District Hospital from March 2000 to February 2001. Information obtained from interviewing caretakers of 165 children with LRI and data collected from medical records revealed the cost per case ranged from 140 to 6,471 baht with an average total cost per case of 1248 baht. The main determinants of the average total cost per case included the diagnosis of diseases, type of patient (outpatient or inpatient), wheezing association and respiratory syncytial virus positivity. Children with severe pneumonia accounted for the highest in the average total cost per case (2,348 baht) while those with bronchitis accounted for the lowest (924 baht). The average total cost per case of inpatients was 3.5 times higher than that of outpatients. Health policy efforts to improve the effectiveness of care in an ambulatory setting may reduce the financial cost of the illness. PMID- 12549802 TI - Wheezing--associated lower respiratory infections in under 5-year-old children: study in Takhli District Hospital. AB - Wheezing was detected in 251 of the 421 (59.6%) children under 5 years old suffering from acute lower respiratory infections (LRI) treated at the Takhli Hospital, Nakhon Sawan Province from November 1998 to October 2000. Bronchitis and pneumonia accounted for 55.0 per cent and 40.6 per cent of LRI with wheezing. Most of the patients, 162 of 251 (64.5%) cases were children under 2 years old. The wheezing symptom declined significantly in children older than 4 years of age. In this study, LRI with wheezing was found all year round but was more frequent in the rainy season similar to other respiratory infections. As the respiratory syncytial virus (RSV) infections were mainly found during the rainy season (July-October), the association of wheezing and RSV infections in the rainy season was evaluated. RSV causing LRI with wheezing in the rainy season revealed 54.7 per cent and 39.4 per cent in the first and second studied year, respectively. The two-year follow-up of 121 cases of LRI with wheezing showed that 37 cases (30.5%) had repeated episodes of wheezing. In conclusion, LRI with wheezing caused by RSV was commonly found in children under 2 years old. The occurrence was all the year round but predominantly in the rainy season. PMID- 12549803 TI - Oral ibuprofen and indomethacin for treatment of patent ductus arteriosus in premature infants: a randomized trial at Ramathibodi Hospital. AB - BACKGROUND: Ibuprofen given intravenously to premature newborn infants is a proven treatment for patent ductus arteriosus (PDA). The efficacy of ibuprofen is comparable to indomethacin in many clinical trials with fewer renal side effects. However, the intravenous form of ibuprofen is not available in Thailand, whereas, the oral suspension form is widely used for antipyretic treatment in children. Therefore, the authors investigated the possibilities of using oral ibuprofen for the treatment of PDA in premature newborn infants. OBJECTIVE: To assess whether oral ibuprofen at 10 mg/kg/dose daily for 3 days was as effective as indomethacin to treat symptomatic PDA in premature infants and to compare the side effects of oral ibuprofen to indomethacin. SUBJECTS AND METHOD: Eighteen premature infants with gestational ages less than 34 weeks born at Ramathibodi Hospital who developed symptomatic PDA were randomly assigned to receive three doses of either indomethacin (oral or intravenous administration 0.2 mg/kg/dose for three doses given at 12 hourly intervals or oral ibuprofen (10 mg/kg/dose for three doses given at 24 hourly intervals). The rates of ductal closure, infants' clinical courses, side effects and complications were recorded. RESULTS: Birth weight, gestational age, gender, age onset and number of infants who had respiratory distress syndrome were similar in both groups, PDA was closed in 7 of 9 infants given ibuprofen (78%) and in 8 of 9 infants given indomethacin (89%) (p > 0.05). The mean plasma concentration of ibuprofen was 28.05 microg/ml at 1 hour after the third dose. Neonates in the ibuprofen group had more urine output. However, the increment of serum BUN and creatinine were not significantly different in both groups. There were no significant differences in duration of ventilator support as well as number of patients with bronchopulmonary dysplasia, intraventricular hemorrhage, necrotizing enterocolitis and death in both groups. CONCLUSION: Oral ibuprofen therapy is as effective as indomethacin for the treatment of PDA in premature infants and seems to have fewer renal side effects. PMID- 12549804 TI - Assessment of pulmonary artery stenosis using freehand "flock of birds" digital color three-dimensional echocardiographic reconstruction. AB - Pulmonary artery (PA) trunk or branch PA stenosis is commonly found in patients with congenital heart disease. The aim of the present study was to evaluate the freehand "Flock of Birds" color Doppler three-dimensional (3D) reconstruction on a modeled-segment imitating PA stenosis. First, a PA model was created from latex tubes to simulate the main PA and its main branches with baseline cross-sectional areas (CSA) of 0.7 cm2. A series of narrowed segments in the right and left PA were created. The cross-sections of the smallest area ranged from 0.13 to 0.59 cm2 and stenotic segmental length ranged from 0.17 to 1.80 cm. The dimensions of these elements mounted on to the model were verified by intravascular ultrasound (IVUS) imaging. Next, pulsatile flows at 60 beats/ min were generated through the system. A GE/VingMed System FiVe with magnetic locator system (Flock of Birds) on a 3.5 MHz transducer was used to acquire a freehand sweep for ECG gated 3D data acquisition of color Doppler flows through the model. The images were reconstructed by EchoPac 3D software and the morphology of the stenotic elements were determined. The results revealed that the narrowest CSA determined by the 3D color flow cast of the pulmonary artery were in excellent agreement with IVUS CSA (r = 0.98, p < 0.001, SEE = 0.04 cm2). The stenotic length estimated from 3D was also in good agreement with the IVUS (r = 0.98, p < 0.001, SEE = 0.03 cm). In addition, complex morphology of the stenosis was well visualized by this technique. As a result, the noninvasive free-hand digital color 3D echocardiography can be adopted for the accurate assessment of the severity and morphology of PA stenosis in patients with congenital heart diseases. PMID- 12549805 TI - The right and left ventricular function after surgical correction with pericardial monocusp in tetralogy of fallot: mid-term result. AB - BACKGROUND: Surgical repair of tetralogy of Fallot (TOF) with reconstruction of the right ventricular (RV) outflow tract invariably results in pulmonary regurgitation (PR). Chronic PR has been associated with RV dysfunction and ventricular arrhythmia. Pericardial monocusp has recently been used at Ramathibodi Hospital to preserve pulmonary valve function. OBJECTIVES: First, to study the competency of the pericardial monocusp, one-year after correction. Second, to assess the right and left ventricular (LV) functions after surgery. Third, to assess correlation between severity of PR and the characters of electrocardiography (ECG) and chest X-ray (CXR) after correction. METHOD: A cross sectional study was conducted in patients who, had undergone total correction for TOF at least one year ago. The past medical history was retrospectively reviewed from the medical records. The patients who underwent surgical correction with and without pericardial monocusp were recruited into group I and group II, respectively. The clinical symptoms, QRS duration from ECG, and cardio-thoracic (CT) ratio from CXR were analyzed. From the echocardiographic standpoint, the LV systolic function was determined by LV fractional shortening (LVFS), whereas the RV systolic function was determined by the tricuspid annular plane systolic excursion (TAPSE). Restrictive physiology of the RV was determined by presence of antegrade flow across the pulmonary valve during diastole. RESULTS: Sixty four patients were enrolled in the study, 7 in group I and 57 in group II. The median follow-up time after the surgery was 6.5 years, which was 3 years in group I and 7 years in group II (p < 0.01). All patients in group I (100%) and 45 (80.4%) in group II had moderate or severe PR. The severity of PR, the RV and LV systolic functions were not statistically significantly different between the two groups (p > 0.01). The median of the LVFS was 32.4 per cent, and of the TAPSE was 10.5 mm. There was no restrictive physiology of the RV in all patients. There were no significant correlations between symptoms, CT-ratio, QRS duration and the severity of PR. CONCLUSIONS: The pericardial monocusp could neither reduce severity of PR nor improve right and left ventricular functions after 3 years follow-up post-operatively. However, the right and left ventricular performances in mid-term period remained insignificantly changed and severity of PR could not be predicted from symptoms and simple laboratory investigations. PMID- 12549806 TI - Sebaceous nevus syndrome as the underlying cause of intractable seizures in a one month-old infant. AB - A one-month-old male infant with generalized seizures since 2 days old was evaluated at the Department of Pediatrics, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand. His seizures were initially characterized as focal movement of the right upper and lower limbs followed by generalized tonic. Initially, phenobarbital was administered but failed to control his seizures. Physical examination revealed generalized mild hypotonia with a hyperpigmented brownish patch affecting the left side of his face. The initial diagnosis was sebaceous nevus syndrome which is one of the neurocutaneous syndromes. The diagnosis was confirmed by skin biopsy of the affected lesion. The literature was reviewed and discussed. The authors emphasized the importance of thorough physical examination including evaluation of specific skin lesion which would be a leading clue in making the diagnosis of symptomatic epilepsy in infants. PMID- 12549807 TI - Cystinuria: cause of recurrent renal stones in a 4-year-old girl. AB - This paper presents the case report of a 4-year and 6-month old girl with cystinuria. She clinically presented with recurrent radiopaque renal stones since the age of 3 years. She received 2 subsequent operations of pyelolithotomy combined with ureterolithotomy at the age of 3 years 6 months, and pyelolithotomy alone at the age of 5 years. She was initially diagnosed as having cystinuria by the presence of hexagonal plate crystals in her acidified urine and positive for the urinary cyanide-nitroprusside test. The diagnosis was confirmed by urinary amino acid analysis using quantitative ion-exchange chromatography which revealed increased amounts of cystine and dibasic amino acids of lysine and ornithine. In spite of maintaining a high fluid intake and alkalinizing urine by giving potassium citrate after the first operation, recurrent renal stones were found. Therefore, after the second operation, D-penicillamine was additionally introduced. During the 18-month follow-up, although there were recurrent renal stones, the rate of stone formation was slower. To the authors' knowledge, this is the first case report in Thailand. PMID- 12549808 TI - Isolated lissencephaly sequence with contiguous gene deletion detected by FISH analysis: a case report. AB - BACKGROUND: Lissencephaly is a clinically and genetically heterogeneous malformation of the brain, usually leading to a severe disabling condition and seizures. The recent discovery of molecular techniques and identification of lissencephaly genes (e.g. LISI and DCX) has allowed etiologic diagnosis of this disorder feasible. OBJECTIVE: To describe a patient with lissencephaly in whom fluorescence in situ hybridization (FISH) determined etiologic diagnosis, providing precise genetic counseling and possible prenatal diagnosis for the family. CLINICAL REPORT AND STUDY RESULTS: The authors report a 4 month-old girl who presented with intractable, generalized myoclonic seizures at I month of age. The patient was born at 37 weeks' gestation, to a G4P1A2 36-year-old woman. Chromosome analysis from amniotic fluid performed for advanced maternal age revealed normal karyotype. Pregnancy was complicated by polyhydramnios. Computed tomographic scan of the brain at age one month showed a total absence of gyral formation. FISH of the metaphase chromosome from the patient, using Smith-Magenis and Miller-Dieker/ILS probe showed two signals of Smith-Magenis probe but only one signal of Miller-Dieker/ILS probe, indicating a microdeletion of 17pl3.3 region including LIS1 gene. Hybridization of the ILS probe on the metaphase chromosome of both parents was normal. CONCLUSION: A confirmation of contiguous gene deletion in this patient lead to an etiologic diagnosis of lissencephaly. This information allowed precise genetic counseling, estimation of recurrent risk, and definite prenatal diagnosis available to the family. The authors suggest FISH 17p13.3 studies be performed in addition to a standard metaphase analysis in all patients with type I lissencephaly. PMID- 12549809 TI - Antineutrophilic cytoplasmic antibody-positive systemic vasculitis associated with propylthiouracil therapy: report of 2 children with Graves' disease. AB - Systemic vasculitis is a rare complication of therapy with antithyroid medication. Antineutrophilic cytoplasmic antibody (ANCA)-associated vasculitis has been described in patients treated with propylthiouracil (PTU) and methimazole (MMI). The majority of cases have underlying Graves' disease. The authors report 2 children who developed ANCA-associated systemic vasculitis during PTU therapy of Graves' disease. One patient, after PTU treatment for 3 years, developed severe systemic vasculitis. After 3 weeks of arthritis, she abruptly presented with hematuria, proteinuria and edema concomitant with anemia. Her serum creatinine was elevated, to 6 mg/dl. Renal biopsy revealed crescentic glomerulonephritis. After admission, she developed intracerebral hemorrhage and pulmonary hemorrhage. She had positive perinuclear-ANCA (p-ANCA) with a titer of 1:160. Despite intensive therapy with immunosuppressive agents and plasmapheresis, as well as discontinuation of PTU, she died of the complications of severe systemic vasculitis. The other patient developed fever, arthralgia and leukocytoclastic vasculitis of the skin during treatment with PTU for about 2 years. Her symptoms and skin lesions disappeared after discontinuation of PTU. However, she has had a persistently high titer of p-ANCA 1:320 through 17 months follow-up time. Thus, patients who are treated with PTU can develop ANCA-positive vasculitis in a mild or severe form. Therefore, they should be carefully followed and monitored, not only for their thyroid status but also the serious complications of PTU. PMID- 12549810 TI - Neonatal lupus erythematosus: clinical manifestations and management. AB - The authors report 6 cases of neonatal lupus erythematosus (NLE) who were seen at Ramathibodi Hospital from 1993 to 2000. The female to male ratio was 1:5. Cutaneous lesions were the major manifestation in all cases. Other clinical manifestations were thrombocytopenia, hepatosplenomegaly and mild elevation of liver enzymes. Skin rashes mostly erupted at 3-6 weeks old. None had a complete heart block but one had abnormal electrocardiograph (ECG) changes compatible with Wolff-Parkinson-White syndrome (WPW). Four of six patients had thrombocytopenia. All of the abnormalities resolved spontaneously except thrombocytopenia. Three of six needed blood transfusion to replace blood loss from gastrointestinal bleeding. Intravenous immunoglobulin (IVIG) 2 g/kg was given in 3 cases with good response in two of three cases. Platelets rose rapidly and maintained at a normal level within 24-48 hours. Combined therapy with corticosteroid 2 mg/kg was given to 1 case with good outcome. Telangiectasia was the most common sequelae especially in patients who had periorbital lesions resembling raccoon's eyes. The authors conclude that IVIG in the dose of 1 g/kg for 1-2 days is an effective treatment for NLE with severe thrombocytopenia especially when corticosteroid is contraindicated. PMID- 12549811 TI - Congenital self-healing Langerhans cell histiocytosis with pulmonary involvement: spontaneous regression. AB - Congenital self-healing Langerhans cell histiocytosis (CSHLCH) is a rare condition which may present at birth or during the neonatal period. It is usually characterized by eruption of multiple, discrete and red-brown papules and nodules which may increase in size and number during the first few weeks of life with spontaneous regression. Systemic signs are usually absent except for occasional mild hepatomegaly. The authors report a case of CSHLCH associated with multiple lung cysts with spontaneous regression. A healthy full term male infant presented at birth with numerous diffuse discrete dark-red papules sized 0.2-1.0 cm scattered on the eyelids, temporal areas of the scalp, neck, palms and soles. Histologic findings were mixed inflammatory infiltration of numerous histiocytes. Immunochemistry findings were S-100 and CDla positive consistent with CSHLCH. Hepatomegaly and multiple lung cysts were detected at 1 month old. Since he was healthy, no medication was given except for close monitoring. At 2 months of age, hepatomegaly was resolved but the liver had sclerosing change. Skin lesions regressed completely at the age of 4 months. Lung cysts were markedly improved at 7 months old and completely resolved at 1 year old. Spontaneous regression in CSHLCH with lung involvement has never been reported in Thailand. PMID- 12549812 TI - Medicaid managed care: are academic medical centers penalized by attracting patients with high-cost conditions? AB - OBJECTIVE: To determine whether case-mix and health utilization disparities exist between Medicaid enrollees within a Michigan managed care organization (MCO) who selected primary care providers (PCPs) affiliated with a major academic medical center (AMC) and enrollees who selected community providers. STUDY DESIGN: A retrospective cohort study using cost estimates obtained from claims data and based on a standardized Medicaid fee schedule. METHODS: We established the prevalence of 25 high-cost chronic medical conditions from the claims data for capitated Medicaid enrollees from January 1, 1997, through October 31, 1999. We assessed differences in healthcare cost estimates per member for Medicaid enrollees at AMC primary care sites versus other community sites using t tests and linear regressions, including analyses stratified for Temporary Assistance for Needy Families (TANF) and Aid to Blind and Disabled (ABAD) programs. RESULTS: Enrollees with AMC providers had a much higher cumulative prevalence of the 25 high-cost chronic medical conditions (95.6 per 1000 enrollees versus 65.6 per 1000; P < .001), and virtually all of this difference was confined to ABAD enrollees. Estimated total costs were also higher for ABAD Medicaid enrollees at the AMC sites than for those at community sites. The average total services and pharmacy cost estimates per ABAD member were $1219 higher per member per year at the AMC sites (P < .001), primarily from costs of inpatient hospitalizations. Regression analyses demonstrated that differences in the prevalence of the 25 high-cost chronic medical conditions accounted for about 50% of the cost differences observed between sites. These analyses suggest that at least half of the observed cost disparity was due to adverse selection. CONCLUSIONS: This study found both significant case-mix and cost disparities for ABAD patients, suggesting that AMC primary care sites experienced substantial adverse selection. Unless approaches to account for adverse selection are put in place, this phenomenon could jeopardize ABAD Medicaid recipients' ongoing access to needed medical care. PMID- 12549814 TI - Effects of ambulatory-care pharmacist consultation on mortality and hospitalization. AB - OBJECTIVE: To determine the effect of 3 alternative models of ambulatory-care pharmacist consultation on patient survival and hospitalization. STUDY DESIGN: Patients were randomly chosen adult Kaiser Permanente (KP) Southern California enrollees with at least 1 prescription in the base year. The demonstration lasted 23 months. Using time-dependent proportional hazards estimation, survival and hospitalization were examined across 5 patient risk groups. INTERVENTIONS: The pharmacy interventions were (1) consultation about new or changed prescriptions as mandated by 1992 state and federal regulations (State model), (2) consultation focused on selected high-risk ambulatory care patients (KP model), and (3) pre 1992 standard care (Control model). RESULTS: In the 6 service areas in which only 1 of the models was implemented, both the KP (relative risk [RR] = 0.295, P < .001) and State (RR = 0.338, P < .001) models significantly reduced emergency hospital admissions over 2 years compared with the Control model when measured across all risk groups. The State model also was associated with fewer urgent and emergency admissions (RR = 0.854, P < .001). In the randomized treatment sample of 5499 patients, the KP model was associated with lower total mortality per new prescription filled (RR = 0.921, P < .01), and significantly lower hospitalization and mortality in high-risk patient groups. CONCLUSIONS: Intensive outpatient pharmacist consultation targeting high-risk patients would improve survival and decrease hospitalization rates. Broader nontargeted consultation would reduce hospitalizations, but was not associated with lower mortality within the 2-year intervention. PMID- 12549813 TI - The cost effectiveness of 5 interventions to increase mammography adherence in a managed care population. AB - OBJECTIVE: To determine the cost effectiveness of 5 combinations of strategies for increasing adherence to mammography recommendations in a population of women between the ages of 50 and 85 years enrolled in a large midwestern health maintenance organization. STUDY DESIGN: A randomized control trial comparing interventions believed to increase mammography adherence. PATIENTS AND METHODS: Intervention strategies included 5 combinations of physician recommendation, telephone, and in-person counseling. A total of 652 participants were randomly assigned to 1 of 6 intervention groups and 628 (95.9%) were available at 6-month follow-up. A logistic regression model with adherence as the dependent variable and group as the independent variable was used to test for significant differences between groups. A ratio of cost to improvement in mammogram adherence evaluated the cost effectiveness at 6 months. RESULTS: All 5 interventions resulted in significantly higher rates of adherence compared to no intervention. However, when considering costs, only 1 emerged as the superior strategy for the overall study population. In-person counseling was the most cost-effective strategy overall, followed closely by in-person counseling and physician's letter intervention. For women contemplating mammography, the telephone contact and physician's letter combination was the most cost-effective intervention. For women not contemplating mammography, the physician's letter was superior. The physician's letter worked best with women who had previous mammograms. For women with no mammogram history, the in-person counseling and physician's letter combination was clearly superior. CONCLUSIONS: The cost effectiveness of mammography screening interventions varies based on women's prior history of mammograms and their future intent. Further, managed care organization member characteristics can be used to determine the most cost-effective mammography screening intervention based on individual readiness. PMID- 12549815 TI - Identifying individuals at risk for the development of type 2 diabetes mellitus. AB - OBJECTIVE: To describe a population-based approach to the identification of individuals at high risk for the development of type 2 diabetes mellitus (DM). STUDY DESIGN: A prospective analysis of the incidence of DM in low- and high-risk groups. PATIENTS AND METHODS: Questions associated with the risk of developing DM were incorporated into a health risk assessment (HRA), and health plan members were invited to complete the HRA as part of worksite health promotion efforts or medical clinic visits. Points were applied to the risk factors for DM, and individual HRA responders were assigned to low- or high-risk groups. A total of 16 427 members completed the HRA. To assess the incidence of new cases of DM, follow-up time was up to 4.8 years (mean, 2.55 years). New-onset DM cases were identified using International Classification of Diseases, Ninth Revision, coding for DM during the timeframe subsequent to HRA completion. RESULTS: The incidence of new DM at the end of the study was 3.5% in the high-risk group and 0.7% in the low-risk group (P < .001). CONCLUSIONS: Identification of individuals at risk for DM using this HRA approach increases by 5-fold the likelihood that the targeted risk reduction efforts will reach the right individuals. This approach allows for resource-efficient targeted intervention efforts designed to reduce the incidence of new DM cases. PMID- 12549816 TI - Impact of an antidepressant management program on medication adherence. AB - OBJECTIVE: To evaluate the impact of mail-based physician and member educational interventions on patient adherence to antidepressant medications. STUDY DESIGN: The randomized controlled prospective design included patients followed for 6 months after filling a new prescription for an antidepressant. A pharmacy claims database was used to identify patients and track medication adherence. PATIENTS AND METHODS: Patients receiving a new prescription for an antidepressant and their prescribers were included. Prescribers were randomly assigned to the intervention and control groups. Patient assignment was linked to their physician's assignment. The control group received no intervention. The educational intervention consisted of monthly letters to patients and prescribers regarding the Health Plan Employer Data and Information Set (HEDIS) standards or educational information regarding the importance of medication adherence. The primary outcome was adherence as measured by the medication possession ratio and measurement as specified by HEDIS. The Student's ttest, the chi2 test, and a logistic regression model were used to compare groups and the variables that affect adherence. Other secondary measurements of adherence were performed. RESULTS: A total of 9564 patients were included. Patients in the intervention group demonstrated greater adherence compared with the control group at 90 and 180 days (P < .05). After adjusting for variables, the intervention variable stood alone in its significant impact on adherence (P <.01; confidence interval, 1.003-1.197). Adherence in the total population was significantly higher for selective serotonin reuptake inhibitors than for other agents (P < .001). CONCLUSION: A monthly mail-based educational intervention program regarding antidepressant medications can positively influence patient adherence to therapy. PMID- 12549817 TI - Efficacy of inhaled corticosteroids administered once daily for asthma. AB - Inhaled corticosteroids (ICS) are the most effective medications for the long term control of asthma. When first used in asthma management, these drugs were scheduled 3- or 4-times daily, but by the late 1980s, it became clear that most patients respond excellently to twice-daily dosing. Numerous recent double-blind, randomized trials have demonstrated the efficacy of once-daily ICS in the management of pediatric and adult asthma. Once-daily dose ICS is most appropriate for patients' mild persistent asthma, and will be effective for many patients with moderate persistent asthma. Some studies suggest late afternoon/early evening dosing is preferable to single daily dosing in the morning. PMID- 12549818 TI - Future directions in protein function prediction. AB - New directions in computational methods for the prediction of protein function are discussed. THEMATICS, a method for the location and characterization of the active sites of enzymes, is featured. THEMATICS, for Theoretical Microscopic Titration Curves, is based on well-established finite-difference Poisson Boltzmann methods for computing the electric field function of a protein. THEMATICS requires only the structure of the subject protein and thus may be applied to proteins that bear no similarity in structure or sequence to any previously characterized protein. The unique features of catalytic sites in proteins are discussed. Discussion of the chemical basis for the predictive powers of THEMATICS is featured in this paper. Some results are given for three illustrative examples: HIV-1 protease, human apurinic/apyrimidinic endonuclease, and human adenosine kinase. PMID- 12549820 TI - Potential protein partners for the N-terminal domain of human topoisomerase I revealed by phage display. AB - Phage display procedure was applied to the N-terminal domain of human topoisomerase I. The consensus sequence identified for clones binding to the N terminal domain was found in 35 human proteins that are either permanently or temporarily located in the nucleus. They are in majority involved in the DNA repair, transcription, RNA metabolism or cell cycle control. Four of identified proteins: Bub3 protein, Cockayne syndrome protein A, damaged DNA binding protein 2 and GRWD protein belong to WD-repeat proteins and their sequences recognized by the N-terminal domain are identically localized. PMID- 12549819 TI - Partial purification and characterization of an 80-kDa transcription factor binding to bHLH motif in the rat p53 promoter. AB - E-box is one of potential cis-regulatory elements for the p53 gene. It was previously reported that USF bound to the E-box of the p53 gene. Recently, we demonstrated that an 80-kDa protein other than USF bound to the E-box and activated the transcription of the p53 gene. In the present study, the 80-kDa protein was partially purified and characterized. First, we confirmed that nuclear factors bound to the E-box in sequence-specific manner by the oligonucleotide competition assay. The binding protein to the E-box was partially purified by a sequence-specific DNA affinity chromatography. The active fraction was analyzed by SDS-PAGE and southwestern blotting assay, which showed that the 80-kDa protein was enriched. The binding activity of the 80-kDa protein was not decreased in the presence of 1.4 M urea. In addition, the binding activity was stable up to 50 degrees C. Treatment of EDTA showed that the 80-kDa protein did not require divalent cation such as Mg2+ for the maximum DNA binding activity. The competition assay with non-specific competitor, poly (dI-dC) showed that the 80-kDa protein had high affinity to its binding site. These biochemical properties provide useful insights into the 80-kDa nuclear factor binding to the p53 promoter. PMID- 12549821 TI - Identification of human CDV-1R and mouse Cdv-1R, two novel proteins with putative signal peptides, especially highly expressed in testis and increased with the male sex maturation. AB - Human systemic carnitine deficiency (SCD) is a hereditary disease caused by the mutation of OCTN2 and has the characteristics of cardiac hypertrophy. Previous studies based on JVS mouse, an animal model of this disease, showed that Cdv-1 was highly expressed in ventricles of normal mouse, but was remarkably down regulated in JVS mouse and can be up-regulated to normal level by breeding carnitine, which suggested Cdv-1 was possibly involved in cardiac hypertrophy caused by carnitine deficiency. In this study, the expression of human CDV-1, a homolog of mouse Cdv-1, was undetectable in heart by northern hybridization. The inconsistent expression levels of human CDV-1 and mouse Cdv-1 in heart implied that cardiac hypertrophy in human SCD might not be associated with the abnormal expression of CDV-1. Interestingly, another long transcripts of the gene, Cdv 1R/CDV-1R, were cloned in the present study, in mouse and human, respectively. This long transcript predominantly expressed in both human and mouse testis and its expression level was increased with testis development. Furthermore, we proved that the open reading frame of Cdv-1R/CDV-1R spans the exons 2 approximately 19 instead of exons 9 approximately 19; and the peptide encoded by CDV-1R was composed of 676 amino acids containing a putative signal peptide instead of 414 amino acids described previously. In addition, it was proved that the expression level of Cdv-1R in JVS mouse testis was as high as that in normal mouse testis, and both were not regulated by carnitine. PMID- 12549822 TI - A novel factor associating with the upstream regulatory element of murine Hoxa-7 induces bacterial cell death. AB - In order to understand the function of a cDNA (c171) associated with the upstream regulatory region of the Hoxa-7, the cDNA was cloned into the pGEX-4T-1 vector to produce it as a GST fusion protein. The size of the fusion protein was determined to be 48-kilodalton (kDa). Sequence analysis revealed that a protein C171 contained one hydrophobic transmembrane domain in the N-terminal region and several putative phosphorylation and glycosylation sites. C171 protein inhibited the bacterial growth within 30 min after induction. The transmission electron microscopic examination revealed that the morphology of the cells expressing C171 was changed dramatically: i.e., unusually elongated phenotype compared with those of controls, and finally leading to a cell death. These results altogether indicate that a trace amount of C171 induces bacterial cell death. PMID- 12549823 TI - Cloning and characterization of a novel human STAR domain containing cDNA KHDRBS2. AB - KHDRBS2, KH domain containing, RNA binding, signal transduction associated 2, is an RNA-binding protein that is tyrosine phosphorylated by Src during mitosis. It contains a KH domain,which is embedded in a larger conserved domain called the STAR domain. This protein has a 99% sequence identity with rat SLM-1 (the Sam68 like mammalian protein 1) and 98% sequence identity with mouse SLM-1 in its STAR domain. KHDRBS2 has the characteristic Sam68 SH2 and SH3 domain binding sites. RT PCR analysis showed its transcript is ubiquitously expressed. The characterization of KHDRBS2 indicates it may link tyrosine kinase signaling cascades with some aspect of RNA metabolism. PMID- 12549825 TI - Isolation and partial purification of a novel type II restriction endonuclease Bsu121 I, from Bacillus subtilis. Bsu121I, a type II restriction endonuclease from Bacillus subtilis. AB - A new type II restriction endonuclease which we designated as Bsu121I has been isolated from gram-positive bacterium Bacillus subtilis strain 121 and partially purified. The restriction endonuclease was isolated from cell extracts using step wise purification through ammonium sulfate precipitation, followed by phosphocellulose column chromatography. SDS-PAGE profile showed denatured molecular weights (23 and 67 kDa) of the endonuclease. The partially purified enzyme restricted pBR322 DNA into two fragments of 3200 and 1700 bp. The endonuclease activity required Mg(+2) as cofactor like other type II endonucleases. PMID- 12549824 TI - Multiple calmodulin genes in fish. AB - In mammals, identical calmodulin (CaM) proteins are encoded by three nonallelic genes that differ in their promoter regions and untranslated regions (UTRs). The UTRs of each of these three genes are specific for each gene and are highly conserved. In this study, sequences obtained from the GenBank and EST databases and sequencing were examined for several species of fish to ascertain whether this multi-gene one protein system exhibited in mammals extends to other vertebrates. Three genes in zebrafish (Danio rerio) designated alpha, beta, and gamma were identified. As in mammals, these genes differ in the 3'-UTR region but encode completely identical CaMs. PCR primers spanning the coding and the 3'-UTR regions were designed based on the assembled sequences and used to confirm the presence of each gene in the cDNA library. Other species of fish were also found to contain homologous genes that were closely related as indicated by phylogenetic analysis. The 3'-UTR of the alpha, beta and particularly the gamma CaM gene of fish were not found to be as conserved as the corresponding genes of mammalian species possibly due to the span of evolutionary time. Only a few short elements in the 3'-UTR were observed to be similar in fish and mammals. These short regions of identity are shared primarily between the mammalian CaM II and CaM I and the alpha gene and beta gene of fish, respectively. Thus, the multi gene one protein system occurs among fish as well as among mammals. PMID- 12549826 TI - Molecular biological characterization of equine surfactant protein A. AB - In the following, we describe the isolation and sequencing of the equine surfactant protein A (Sp-A) as found in both the cDNA and the genomic DNA. We found a length of the cDNA sequence of 747 bp (base pairs), in translation into amino acids of 248. Compared with the known molecular biological facts about Sp-A in other species, the cDNA sequence obtained showed highest homology with that of sheep (85.01%). The genomic DNA of equine Sp-A, as in other species, includes three introns. There were no hints for the existence of two different Sp-A genes. These results should form the basis for a better understanding of respiratory failure in foals and adult horses, and also lead to further studies on this item. PMID- 12549827 TI - Reference values for the bone marrow aspirates in adult dogs. AB - Reference ranges for each cell type and six different ratios (M : E ratio = quotient of number of myeloid cells/erythroid precursors; different maturation ratios) were calculated for bone marrow aspirates of adult dogs. These values were based on 2.5 and 97.5% percentiles of differentials of 1000 cells in bone marrow aspirates of 92 healthy 1-8-year-old dogs. The results of intact male and female dogs were compared. No distinct sex-related differences were found (P > 0.05). A wide physiological range was observed in almost all bone marrow cells resulting in broad reference ranges for the ratios. The clinically relevant M : E ratio varied between 0.45 and 2.87. The accurate cytological examination of bone marrow based on the reference ranges presented in this study requires preparation of high-quality bone marrow films with minimal blood contamination. PMID- 12549828 TI - Laboratory changes consistent with feline infectious peritonitis in cats from multicat environments. AB - The present study describes the prevalence of haematological and electrophoretic changes consistent with the diagnosis of feline infectious peritonitis (FIP) in cats without FIP living in six multicat environments with different prevalence of FIP and of other diseases. The results allow designing haematological and electrophoretic profiles typical of each group, most likely depending on the management and on the health status of the group rather than on the prevalence of FIP. In fact, many cats from the colonies with open management and frequent outbreaks of infectious diseases other than FIP had one or more haematological and/or electrophoretical changes consistent with FIP, compared with the reference ranges. In the case of non-specific clinical signs such as fever or neurological signs because of diseases other than FIP, these cats would be erroneously considered as affected by FIP and euthanasized. The use of internal ranges designed on the basis of repeated samplings from non-symptomatic cats allows avoiding these misinterpretations. Results from cats with symptoms consistent with FIP living in the same colonies were also compared with both the reference ranges and the internal ones: such a comparison demonstrated that the use of internal ranges rarely affected the possibility to correctly diagnose the disease in cats with symptoms suggestive of FIP. PMID- 12549829 TI - Ultrasonographic observations of the uterine regression in the ewe under different obstetrical conditions. AB - Involution of the ovine uterus after parturition was studied using real-time B mode ultrasonography. The diameter of the uterus, the uterine lumen and the caruncles were investigated during the first 30 days post-parturition (p.p.) by transrectal and transabdominal application of the ultrasound probe. Ewes that spontaneously delivered showed a physiological regression of the uterus with a transversal diameter of 4.9 (+/- 0.86) cm on day 1 p.p. which decreased to 1.84 (+/- 0.14) cm until day 30. The course of uterine regression was established on these data and was described by the exponential equation: y = E + A * e(-b*t). 'E' means the final value on day 30 p.p. and 'A' the difference between the initial transversal diameter on day 1 p.p. minus the final value on day 30 p.p. The exponents 'b' and 't' indicate the downward gradient of the regression curve and also the factor time in days. Furthermore, we found that the uterine regression in ewes finishes approximately on day 17 p.p. The uterine involution was delayed in ewes after manual obstetrics and caesarean section, in addition the incidence for retention of foetal membranes was increased. The present study demonstrates that the ultrasonographic technique is a suitable method for the control of the uterine involution and for the diagnosis of pathological aberrations in sheep. PMID- 12549830 TI - Evaluation of the pathology, pathogenesis and aetiology of auricular elephantiasis in slaughter pigs. AB - Ears from slaughter pigs with auricular elephantiasis (n = 24) and the corresponding lymph nodes (lnn.) (n = 26) were grossly, histopathologically and microbiologically examined. Immunostaining for IgM, IgG, Cd3epsilon and bacterial antigens of Arcanobacterium pyogenes and Staphylococcus aureus was performed by indirect enzyme-based techniques. Ears were variably thickened depending on the sampled area (basis, centre and apex). However, at all locations the thickness, the length from basis to apex and the weigh of whole ears with elephantiasis were significantly increased (P < 0.01). The corresponding lnn., that is, ln. parotideus superficialis and profundus, had also increased significantly (P < 0.01) in volume. Histopathologically, lesions of the ears and the corresponding lnn. revealed changes characterized by diffuse fibrosis intermingled with multiple pyogranulomatous foci containing asteroid bodies. In the majority of lesions, four distinct zones due to different cellular infiltrates encircled the central core of the asteroid bodies. In several lesions, the pyogranulomatous foci were contained within the lymph vessels. Immunohistochemically, only the bacterial antigen of S. aureus was detected within the cytoplasm of the macrophages and/or in the asteroid bodies of the ears (41.5%) and in the regional lnn. (30.8%). An abundant number of IgM, IgG and CD3epsilon-positive cells were present in all the pyogranulomatous lesions, whereas a positive IgG-staining was observed only in a single asteroid body. Thus, porcine auricular elephantiasis is a chronic pyogranulomatous inflammation that is frequently positive for S. aureus and is lymphogenically spread. Therefore, the lesions of the ears with auricular elephantiasis and the corresponding lnn. should be termed auricular botryomycosis and botryomycotic lymphadenitis, respectively. Moreover, as the disease is observed frequently in slaughter pigs it must also be considered according to the welfare of the animals and in relation to post-mortem meat inspection. PMID- 12549831 TI - Cerebellar granuloprival degeneration in an Italian hound. AB - A severe atrophy of the cerebellum was observed in a 7-month-old male Italian hound with a history of progressive ataxia and head tremor from the age of 3 months. On clinical examination, signs included severe hypermetric gait, head tremors and proprioception deficits in all limbs. At necropsy, a pronounced symmetrical reduction in size of the cerebellum was the only gross lesion observed. Histological examination of the cerebellum revealed marked thinning of the granular and molecular layers with almost complete loss of granule cells. Purkinje cells had normal morphology and distribution. These findings differ from those of previous reports of cerebellar cortical abiotrophy in dogs, which were mainly characterized by prominent Purkinje cell degeneration and loss. PMID- 12549832 TI - Central nervous system cryptococcoma in a cat. AB - This report describes the gross, histopathological, immunohistochemical and electron microscopic findings in a 4-year-old cat with systemic Cryptococcus neoformans infection. A 1-cm diameter pontine mass, pinpoint lesions in the cerebellum and in the right kidney were the main macroscopic findings. A presumptive diagnosis of cerebral neoplasia with metastasis was formulated. Light microscopy revealed a huge number of yeasts surrounded by a pyogranulomatous inflammatory reaction in the cerebral parenchyma and, to a lesser extent, in the kidney, while the meninges were not involved. The positive mucicarmine stain that coloured the capsule of the yeasts was indicative of cryptococcal infection. Immunohistochemistry confirmed the presence of C. neoformans var. grubii (C. neoformans serotype A). Electron microscopy revealed yeasts with a massive fibrillar capsule and lamellar cell wall free in the cerebral tissue and within macrophages. PMID- 12549833 TI - Effect of lead exposure on the erythrocytic antioxidant levels in goats. AB - Fifteen adult female goats were orally exposed to 5.46 mg lead (as lead acetate) per kg body weight daily for 2 weeks to study the antioxidant enzymes of the erythrocyte, lipid peroxide level, total thiol groups and total antioxidant status (TAS) in plasma. Ten goats served as unexposed control. Blood samples were collected before exposure (day 0) and on days 7 and 14. Ten per cent erythrocyte haemolysate was prepared and analysed for glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT), total thiol groups and lipid peroxide. TAS was determined in plasma. There was a significant (P < 0.05) increase of erythrocytic GPx, SOD, CAT, total thiol groups and TAS on day 7 which was followed by a significant (P < 0.05) decrease of all these parameters on day 14. Lipid peroxide level increased significantly (P < 0.05) and the maximum level was attained by day 14. The results obtained indicate a possible role of free radicals in lead poisoning pathogenicity. PMID- 12549834 TI - Sertoli cell tumour with Call-Exner-like bodies in a dog. AB - An histological diagnosis of Sertoli cell tumour and concurrent seminoma was formulated after complete resection of the right testicle in a 8-year-old Doberman. The unusual histological feature of abundant eosinophilic material with a extracellular location forming the Call-Exner-like bodies is reported. PMID- 12549835 TI - Metastasizing fibrosarcoma in a Wistar rat--case report. AB - A 19-month-old male control Wistar rat from a 30-month inhalation study showed a subcutaneous greyish-white mass extending from the throat to the thoracic cavity. The rat had been euthanized because of its poor general condition. Histologically, the mass was diagnosed as a fibrosarcoma infiltrating the masseter muscle with metastases in the lungs, liver and heart. The primary tumour was characterized by fusiform spindle cells producing various amounts of interlacing bundles of collagen. The cells formed a characteristic herringbone pattern and mitotic figures were frequent. The histological parameters of the metastases were practically identical to those seen in the primary tumour. The diagnosis was confirmed by trichrome staining and positive immunostaining for vimentin and was differentiated from leiomyosarcomas by its negative immunostaining for desmin, from schwannomas by its negative immunostaining for S 100 and from malignant fibrous histiocytomas by the absence of giant cells. The incidence of fibrosarcomas in Wistar rats is very low (up to 3%) and metastasis is rarely observed. PMID- 12549836 TI - Hepatocholangiocellular carcinoma in a rat--case report. AB - A mixed epithelial tumour in the liver of a 24-month-old male Wistar rat from a 30-month inhalation study is described. The rat, which was in a group exposed to low concentrations of diesel exhaust, was euthanized because of emaciation, forced respiration and abnormal gait. Macroscopic examination of the enlarged liver revealed multiple partly confluent beige-red nodules up to 1.5 cm in diameter. Small nodules up to 7 mm in diameter were seen in the spleen. Histologically, the tumour nodules in the liver consisted of hepatocellular and cholangiocellular components. The hepatocellular component consisted of moderately differentiated polygonal to round hepatocytes about twice as large as normal hepatocytes and having hyperchromatic, centrally located nuclei with prominent nucleoli and eosinophilic cytoplasm. Foci of haematopoiesis and focal necroses were prominent. The cholangiocellular component was moderately differentiated and consisted of tubular structures lined by low cuboidal to cylindrical cells showing cytoplasmic basophilia and small dark nuclei without prominent nucleoli. The histological features of the nodules in the spleen corresponded to those of the primary tumour in the liver. Based on these criteria, the tumour nodules were diagnosed as hepatocholangiocellular carcinoma. The immunohistological examination confirmed the diagnosis, i.e. immunostaining for cytokeratins was positive for eight and 18 (hepatocellular carcinoma) and for seven and 19 (cholangiocellular carcinoma) as well as for vimentin (dense fibrous stroma). This tumour is considered to be spontaneous because of its single occurrence. PMID- 12549837 TI - Accompanied MDT (AMDT)--more questions than answers. PMID- 12549838 TI - Can social marketing be applied to leprosy programmes? AB - The implementation of multidrug therapy (MDT) has been highly effective in curing patients and reducing leprosy prevalence. In some countries, however, a significant number of cases remain undetected or are detected late. Although compliance with drug therapy is generally good, a significant proportion still defaults treatment in countries where the leprosy burden is still high. This paper proposes that leprosy control or elimination efforts might be enhanced by the application of social marketing principles. It first outlines the principles of social marketing and then reviews a successful social marketing campaign in Sri Lanka to increase case detection and treatment. The paper concludes with a discussion of the opportunities for using social marketing principles to enhance the success of current leprosy community heath education programmes and leprosy treatment services. PMID- 12549839 TI - Tendon transfer for triple nerve paralysis of the hand in leprosy. AB - Paralysis of ulnar, median and radial nerves is seen in less than 1% of those affected with leprosy. This condition is a particular challenge for the surgeon, physiotherapist, and patient. A retrospective chart review was conducted at the Green Pastures Hospital and Rehabilitation Centre (GPHRC) and Anandaban Leprosy Hospital (ALH) in Nepal, and results were graded by the system outlined by Sundararaj in 1984. Thirty-one patients were identified, and 21 charts were available for review. Excellent or good results were obtained in 93% of patients for wrist extension, 85% of patients for finger extension, 90% of patients for thumb extension, 71% of patients for intrinsic reconstruction, and 63% of patients for thumb opposition reconstruction. These results are reasonable but inferior to those obtained by Sundararaj in his study. Surgical intervention offers a very significant improvement in function in these very difficult hands. Intensive physiotherapy is required both pre- and postoperatively. PMID- 12549840 TI - Leprosy in Mpumalanga Province, South Africa--eliminated or hidden? AB - In South Africa, leprosy has been a notifiable condition since 1921. Although the WHO elimination target of less than one case per 10,000 population has been achieved at country level, the distribution of leprosy in the country is distinctly heterogeneous, with a prominent 'leprosy belt' of greater prevalence stretching across Mpumalanga Province into northern Kwa-Zulu Natal. The highest prevalence in this 'belt' has historically been in Ermelo District. Recent trends of few newly detected leprosy patients in this district raised concerns that health system changes may have resulted in failure to detect leprosy cases. Thus a large-scale community awareness campaign was conducted followed by an intensively advertised screening programme of 3-month duration at schools and central gathering points in villages and farms from 1 June to 31 August 2000. One thousand one hundred and seventy-seven people presented for clinical screening at designated points, while 790 scholars were screened at schools and an additional 1433 people were screened at their homes by the field team. Forty-four people with skin or nervous system lesions compatible with leprosy were referred for specialized assessment and biopsy where indicated. Four new leprosy patients were diagnosed, including an elderly lady with pronounced disability. Two of these patients had prior contact with the health service due to dermatological manifestations of leprosy without diagnosis being made. All patients provided a history of close prolonged contact with known leprosy patients. Ongoing intense tracing and follow-up of close contacts of proven leprosy cases may be a more efficient method of detecting leprosy cases in areas with relatively stable populations that have accomplished 'leprosy elimination', than resource intensive community surveys. PMID- 12549841 TI - Is beggary a chosen profession among people living in a 'leprosy colony'? AB - Leprosy is a highly stigmatized disease that apart from the physical ailments and the deformities causes psycho-socio-economic problems to the people affected. As a result of social rejection, leprosy colonies were formed inhabited by the leprosy-affected families. With inadequate socioeconomic support and help, these people often have resorted to beggary as a way to earn their living. This study is an attempt to look into the lives of the leprosy-affected people living in the leprosy colony in Ambala City, Haryana, north-west State of India and who have accepted beggary as their source of income. The psychosocial impact of leprosy and the subjects' attitude towards beggary has been studied. The study comprised 21 families, including, 22 men, 21 women and 40 children. Seventy-one percent of the families came from Southern India. All the men and nine of the women were leprosy-affected. The proportion of people with deformity was 89%. Prior to contracting leprosy, all of the men were employed, mainly in agriculture and physical labour. At present, all are beggars. Of the 20 who were interviewed, 65% of those who beg and 83% of other adults were illiterate. Fifty percent of the children were in need of education. Due to leprosy, the social interaction of 85% of the interviewees was limited to within the colony and of 88% to only other leprosy-affected people. Through their own organized efforts, they raised welfare services and housing for themselves. None of them liked begging to start with but have accepted it as a source of income. If given a chance and support, 80% said they were ready to quit begging. They were concerned about the education of their children. The study highlighted the need to develop alternate avenues of income generation utilizing the existing desires and potential of the inhabitants. PMID- 12549842 TI - Dehabilitation of leprosy-affected people--a study on leprosy-affected beggars. AB - Leprosy is one of the most socially stigmatized diseases known today. Social stigma is associated mainly due to the prevalent myths like its hereditary and contagious nature, divine curse along with the physical deformities caused. The affected people not only face physical impairments but also suffer psychosocial repercussions due to the community's attitude. The long-term physical and psychosocial restrictions slowly push the leprosy-affected person out of the society. With lack of social support and self-confidence, some dehabilitated leprosy-affected persons end up as beggars. The present study focuses on the long term consequences of leprosy. It is based on case studies of leprosy-affected beggars in Delhi. The process of dehabilitation in each case has been studied. It has been found that dehabilitation is a continuous process. The combination of leprosy, physical impairments and social stigma causing further participation restriction, lead to dehabilitation of people affected by leprosy, and ending in a state of beggary for some. There is a need to develop a holistic approach including both prevention of dehabilitation and rehabilitation of those dehabilitated to overcome both the disease and its consequences. Measures to prevent such dehabilitation in future along with the rehabilitation of leprosy affected beggars have been suggested. Both these measures should take place simultaneously. PMID- 12549843 TI - Sociocultural considerations in the treatment of leprosy in Rio de Janeiro, Brazil. PMID- 12549844 TI - A comparison of Rapid Village Survey and Leprosy Elimination Campaign, detection methods in two districts of East Java, Indonesia, 1997/1998 and 1999/2000. AB - A Rapid Village Survey (RVS) was planned to estimate the extent of the leprosy problem in two well documented endemic districts of East Java, Indonesia. Furthermore, the aim was to investigate the efficacy of the routine programme in detecting new and early cases, as well as the feasibility of RVS in detecting disabled people affected by leprosy in the community. A random sample survey (RVS: a simple method compared to a Population Sample) was used to determine the extent of the leprosy problem. In addition, a Leprosy Elimination Campaign (LEC), was used particularly to detect new and backlog cases in the community. Both RVS and LEC involve a health education campaign followed by the examination of persons voluntarily reporting. Routine programme case finding, involving passive case finding and contact examinations, was also carried out. The RVS prevalence rate of 12 per 10,000 was more than twice the known prevalence rate of 5 per 10,000. The LEC prevalence rate was less than the rate found by RVS, but was within the RVS confidence interval. During the RVS, many children with leprosy were detected, and 10% of all RVS new cases already had disability grade II. The population disability grade II rate due to leprosy was 9 per 10,000. Despite the fact that an active leprosy control programme had been carried out in the surveyed endemic area over a period of many years, the actual prevalence rate found was more than twice the known prevalence. Many children were found during the RVS, thus indicating continuing widespread transmission. In general, it seems that there is still a serious delay in detecting new cases under the routine programme. Consequently, there are substantial numbers of persons affected by leprosy in those districts in need of rehabilitation. PMID- 12549845 TI - The impact of HIV on tuberculosis control--towards concerted action. AB - The well-established international control strategy for tuberculosis is based upon passive case-finding of the most infectious cases followed by effective chemotherapy with sufficient support to ensure and record a successful outcome. However, no country with a severe HIV epidemic is successfully controlling tuberculosis. HIV exerts a double blow. Not only must the health service manage a greatly increased number of patients (as many as fourfold higher in many African settings) but each individual patient needs to be managed more effectively if the control programme is to have a similar impact on transmission as it did in the pre-HIV era. In this paper, we discuss some of the effects of increased burden and stigmatization. We consider the potential of preventive therapy to reduce the impact of HIV on tuberculosis control and describe a more integrated approach to both infections that is being piloted in several sites in Southern Africa. PMID- 12549846 TI - Relapse of lepromatous leprosy after WHO/MDT with rapid bacterial growth. AB - The authors report a case of relapse in a lepromatous patient 6 years after he had been cured by MDT/WHO/24 doses. The atypical aspect emphasized in this case is the bacterial load increase in a short period of time of 1 year after the smear count was negative, and the case reinforces the importance of patient education on release. No leprosy cases were identified in the patient's close contacts. It seems that relapse was a result of bacillary persistence, since a significant improvement was noted in relapsed lesions after two doses of MDT/WHO. PMID- 12549847 TI - Ballpoint pen testing: light touch versus deep pressure. PMID- 12549848 TI - Integration of leprosy services. PMID- 12549849 TI - Leprosy bug provides clues to early nerve degeneration. PMID- 12549850 TI - Bioinformatics in disease endemic countries. PMID- 12549851 TI - Rehabilitation in China. PMID- 12549852 TI - Ayurveda--the people's medicine. PMID- 12549853 TI - Update on the fight against lymphatic filariasis. PMID- 12549854 TI - Wnt signaling in the vasculature. AB - The Wnt signaling pathway regulates normal development as well as a variety of pathologies. Studies of the Wnt pathway have focused largely on very early development and on tumorigenesis. Recent observations point to a role for Wnt signaling in vessel development and pathology. Although not yet investigated systematically, several Wnt ligands have been demonstrated to be expressed in the cells of blood vessels in vivo and in vitro, including Wnt-2, -5a, -7a and -10b. Mice deficient for Wnt-2 display vascular abnormalities including defective placental vasculature. Wnt receptors, called frizzled (Fz), are also expressed by vascular cells in culture and in situ. Of the 10 murine Fz identified to date, Fz 1, -2, -3, and -5 have been demonstrated in endothelial and vascular smooth muscle cells; mice deficient for Fz-5 display vascular abnormalities and are embryonic lethal. Two soluble, naturally occurring Wnt antagonists, frizzled related proteins (FRP)-1 and -3, are also expressed by vascular cells. Stabilization of the downstream signaling component beta-catenin in blood vessels has been demonstrated in several developmental and pathologic states, further supporting the idea that Wnt signaling plays an important regulatory role in the vasculature. PMID- 12549855 TI - Long-term survival of a patient with small-cell lung cancer (SCLC) following treatment with thalidomide and combination chemotherapy. AB - Thalidomide, an angiogenesis inhibitor is currently used in clinical trials in the US and Europe in combination with chemotherapy for the treatment of various solid tumors. The prognosis of patients suffering from small-cell lung cancer (SCLC) is poor. A 73-year-old female with extensive disease of SCLC was given six courses of chemotherapy with adriamycine, cyclophasphamide and oncovine, which led to complete remission of the disease. Following written informed consent, the patient has been treated with thalidomide 200 mg orally on a daily basis for 2 years and 5 months. There has been no sign of tumor recurrence during the follow up. This case underlines the possible role of additional treatment with angiogenesis inhibitors in combination with traditional chemotherapy in the therapy of SCLC. Although there is no proof that thalidomide contributed to this good outcome and no conclusions can be drawn from this treatment in a single patient, further studies may determine the role of thalidomide as an adjuvant antiangiogenic agent in the therapy of SCLC. PMID- 12549856 TI - A computer image analysis system for microvessel density measurement in solid tumours. AB - Microvessel density (MVD) is a widely used surrogate measure of angiogenesis in pathological specimens and tumour models. Measurement of MVD can be achieved by several methods. Automation of counting methods aims to increase the speed, reliability and reproducibility of these techniques. The image analysis system described here enables MVD measurement to be carried out with minimal expense in any reasonably equipped pathology department or laboratory. It is demonstrated that the system translates easily between tumour types which are suitably stained with minimal calibration. The aim of this paper is to offer this technique to a wider field of researchers in angiogenesis. PMID- 12549857 TI - Human vascular endothelial cells with extended life spans: in vitro cell response, protein expression, and angiogenesis. AB - An in vitro angiogenesis system was designed for screening angiogenic agonists and antagonists. In order to obtain large quantities of cells and reproducibility, human endothelial cells with extended life spans were developed by retroviral transfection. The resulting cells grown in a serum-free medium containing endothelial cell growth supplement (ECGS) have a telomerase activity, extended life spans of at least 21 passages, and an endothelial cell phenotype (diI-acetylated-LDL upake, factor VIII-related antigen, VEGFR-1 and R-2, and tissue-type plasminogen activator (tPA)) that resembled that of unaltered primary endothelial cells. Exceptions were (i) a higher expression of tPA, and (ii) a non significant growth response to FGF-2 or VEGF stimulation. Within three dimensional fibrin gels, specific cell clones rapidly formed tubular structures in a more reproducible manner than those observed with low-passage primary cells. Tube formation by primary endothelial cells and those with extended life spans was dependent upon FGF-2 and ECGS, respectively. Both cell types produced FGF-2 and VEGF cytokines. Increasing doses of suramin significantly decreased the size of microvessels formed by both cell lines. These functional results indicate that a vascular matrix system containing human cells with extended life spans can be successfully utilized as an in vitro assay for antiangiogenic compounds. PMID- 12549858 TI - A monoclonal antibody that blocks VEGF binding to VEGFR2 (KDR/Flk-1) inhibits vascular expression of Flk-1 and tumor growth in an orthotopic human breast cancer model. AB - Vascular endothelial growth factor (VEGF) is a primary stimulant of tumor angiogenesis. We previously raised a neutralizing anti-VEGF monoclonal antibody 2C3 that blocks the interaction of VEGF with VEGFR2 (KDR/Flk-1) but not with VEGFRI (FLT-1/flt-1). Here, we describe the therapeutic effects of 2C3 on tumor growth in an orthotopic model of MDA-MB-231 human breast carcinoma implanted in the mammary fat pads (MFP) of nude mice. Administration of 2C3 to mice with 100 150 mm3 tumors inhibited tumor growth by 75%, as compared to recipients of the isotype-matched irrelevant control IgG, C44. Treatment with 2C3 also inhibited the establishment of tumor colonies and reduced tumor burden in the lungs of mice injected intravenously with MDA-MB-231 cells. No toxicity was observed in these studies. The mean microvascular density (MVD) of tumors in 2C3-treated mice was 55 +/- 5 per mm2, as compared to 188 +/- 5 per mm2 in the C44-treated control group. The decrease in MVD closely correlated with the degree of inhibition of tumor growth. Treated tumors mostly contained mid-size and large vessels. Microvessels were mainly confined to the peripheral layer of tumor that bordered on the normal MFP epithelium. Tumor vessels had decreased expression of VEGFR2, indicating that neutralization of tumor-derived VEGF by 2C3 down-regulates the expression of VEGFR2 on tumor vasculature. This, in turn, may limit reinitiation of angiogenesis by either tumor-derived or stromal VEGF. These findings suggest that 2C3 is a candidate for treating primary cancer and for preventing the outgrowth of tumor metastases in cancer patients. PMID- 12549859 TI - Comparative study of isoflavone, quinoxaline and oxindole families of anti angiogenic agents. AB - A study designed to compare the effects on VEGF-induced angiogenesis of a number of known anti-angiogenic agents together with some novel derivatives thereof was undertaken. Thus the isoflavone biochanin A 1[structure: see text], indomethacin 2[structure: see text], the 3-arylquinoxaline SU1433 and its derivatives 3 6[structure: see text], the benzoic acid derivative 7[structure: see text], the oxindoles SU5416 8[structure: see text] and SU6668 11[structure: see text], together with their simple N-benzyl derivatives 9, 10, and 12[structure: see text] were selected for study. Using an in vitro assay the compounds were evaluated for their ability to inhibit VEGF-induced angiogenesis in HUVECs, and the cytotoxicity of representative compounds was also studied in tumour cell lines using 24-h exposure. The results indicate that the SU compounds, SU1433, SU 5416 and SU6668, are more potent inhibitors of VEGF-induced angiogenesis than indomethacin or the naturally occurring biochanin A, presumably because they inhibit VEGF receptor signalling. Blocking one of the phenolic OH groups of SU1433 reduced anti-angiogenic activity, as did blocking the NH groups of SU5416 and SU6668. Cytotoxicity studies indicate that none of the compounds examined exhibited cytotoxicity at anti-angiogenic concentrations. PMID- 12549860 TI - Strain differences and inheritance of angiogenic versus angiostatic activity in oestrogen-induced rat pituitary tumours. AB - In this study we investigated the control of the angiogenic/haemorrhagic phenotype of oestrogen-induced rat pituitary tumours of an F1 hybrid (F1) of Fischer 344 (F344) (tumour susceptible) and Brown Norway (tumour resistant) strains. F1 forms a pituitary tumour upon chronic oestrogen treatment, but microvessel count (MVC) is no greater than untreated. In other words, F1 MVC keeps pace with tissue growth during growth of an oestrogen-induced tumour. On the other hand, F344 showed a significant increase in MVC (P = 0.002) upon chronic oestrogen treatment during growth of a large pituitary tumour. F1 control vasculature showed features intermediate between the parent strains, while oestrogen-treated F1 pituitary has pronounced changes commensurate with vascular remodelling or angiogenic activity, along with regressive changes. In addition, oestrogen-treated F1 does not form the haemorrhagic lakes characteristic of oestrogen-treated F344. We conclude that F1, which has a 50-50 genetic composition of the tumour susceptibility and tumour resistance alleles, shows loss of angiostatic activity in the absence of an effective angiogenic stimulus. As a result it is unable to make the 'switch' to the angiogenic phenotype. PMID- 12549861 TI - Inflammatory infiltrate, VEGF and FGF-2 contents during corneal angiogenesis in STZ-diabetic rats. AB - Angiogenesis is a key mechanism that influences several physiological and pathological processes, including wound healing. During the past decades, many groups have shown that controlling angiogenesis might be an answer to overcome pathological situations when this process is out of control. Many altered metabolic states exert considerable influence on the development of angiogenesis. We have chosen diabetes as a model of a progressive metabolic disease with many associated conditions, including an alteration of wound healing dynamics described elsewhere. To evaluate the growth of newly formed blood vessels during diabetes, we induced corneal angiogenesis through silver nitrate cauterization in streptozotocin-induced diabetic rats, always comparing to control non-diabetic or insulin-treated diabetic rats. Computer-aided analysis showed that both the percentage of area taken by vessels on the cornea and their average length were decreased in diabetic animals; furthermore, this diminishment was prevented by insulin treatment in previously diabetic rats. Immunohistochemical staining of neutrophils and macrophages (EDI clone) did not show any differences on number of migrating cells in the cornea. Immunolocalization of vascular endothelial growth factor and basic fibroblast growth factor did not differ considerably among groups either. These results support previous findings that angiogenesis is decreased due to the development of diabetes mellitus but contrasts to descriptions from other investigators in regard to the inflammatory infiltrate and production of growth factors. In our experimental conditions, the cause of the decreased angiogenesis in diabetic rats remains for further elucidation. PMID- 12549863 TI - The thin prep rat aortic ring assay: a modified method for the characterization of angiogenesis in whole mounts. AB - The rat aortic ring model has gained broad acceptance as an angiogenic assay. This system can be used to study the activity of angiogenic and anti-angiogenic factors, and investigate the molecular mechanisms of the angiogenic process. We describe here a thin prep modification of the aortic ring model, which significantly simplifies the procedure and allows staining of aortic outgrowths as whole mounts. Using this procedure, intact preparations of angiogenic outgrowths are successfully and reproducibly stained with endothelial cell (anti CD-31 and -Tie2 antibodies, Griffonia Simplicifolia isolectin-B4) and smooth muscle cell (anti-alpha-smooth muscle actin antibody) markers. Combined use of double immunostaining and confocal microscopy allows concurrent visualization of endothelial and mural cells in the same cultures. Whole mount immunostains of rat aorta cultures are an effective way to rapidly characterize the cellular composition of the angiogenic outgrowths, and localize proteins implicated in the regulation of angiogenesis. This method should facilitate the work of the many vascular biologists that have adopted the rat aorta model as a tool to study angiogenesis and its mechanisms. PMID- 12549862 TI - The sponge/Matrigel angiogenesis assay. AB - It has become increasingly clear that definitive tests for angiogenesis require in vivo assays. Recently, the Matrigel plug assay has become the method of choice for many studies involving in vivo testing for angiogenesis. In this assay, test angiogenesis-inducing compounds such as bFGF or tumor cells are introduced into cold liquid Matrigel which, after subcutaneous injection, solidifies and permits penetration by host cells and the formation of new blood vessels. Assessment of angiogenesis in the Matrigel plug is achieved either by measuring hemoglobin or by scoring selected regions of histological sections for vascular density. We now describe a modification of the Matrigel plug assay which permits a more precise visualization of the angiogenic reaction, provides directional information, requires no histological analysis, and lends itself to photographic documentation and image analysis protocols. We illustrate the assay by describing dose- and time-dependent responses to tumors of murine and human origin, to angiogenesis inducing factors such as bFGF (FGF-2) and VEGF and to anti-angiogenic agents such as endostatin. The method has been used as well to demonstrate blood vessel recruitment by embryonic chick aortic arch rudiments. Additionally it has been able to detect strain-dependent differences in susceptibility to angiogenic stimulation. PMID- 12549864 TI - Restoration of blood flow and evaluation of corresponding angiogenic events by scanning electron microscopy after a single dose of VEGF in a model of peripheral vascular disease. AB - The angiogenic effect of vascular endothelial growth factor (VEGF) has typically been assessed by indirect methods, including microsphere injection and angiography. Here, we use 3-D scanning electron microscopy (SEM) to directly visualize patterns of angiogenesis after a single bolus administration of VEGF in a model of peripheral vascular ischemia. Hind limb ischemia was induced by subcutaneous turniquet implantation in adult Wistar rats. The control group (group A, n = 10) was left untreated, group B (n = 10) received a single dose of VEGF (50 microg) injected in the peroneus muscle. LASER Doppler was applied for blood flow measurements. Animals were sacrificed on day 14 after ischemia induction and vascular casting was performed. Angiogenetic events such as 'tiny lateral sprouts', arcus formations', confluences and the angle of sprouting were assessed by SEM. Significant capillary sprouting was observed in both groups. VEGF-treated limbs demonstrated higher degrees of capillary growth (P = 0.01) and flow recovery (P = 0.028). 3-D-SEM showed sprouts to be more frequent in group B. Tiny lateral sprouts, which always left the mother vessel at an angle of 90 degrees and which were of small diameter and lacked imprints of endothelial cell nuclei, were more frequent in the VEGF-treated group (P = 0.018). Arcus formation was significantly higher in the treated group (P = 0.02). We have developed a simple and effective experimental model of ischemia. For the study of angiogenic phenomena, 3-D imaging of the microvasculature offers a direct and conclusive method for the study of angiogenic events. PMID- 12549865 TI - Protease activated receptor-1 is down regulated by levonorgestrel in endometrial stromal cells. AB - Progestogens are used clinically for contraception, to control excessive menstrual bleeding and to oppose oestrogen in hormone replacement therapy. The use of intrauterine levonorgestrel (LNG) is however, associated with endometrial atrophy and decidualisation of the stroma. In this study, we aimed to identify genes whose expression is modulated by LNG either alone or in combination with progesterone. Thus endometrial stromal cells were stimulated with progesterone, LNG or LNG and progesterone. Poly-A RNA was isolated and used to probe expression arrays. The expression of a number of genes was altered on exposure to LNG or LNG and progesterone. Alteration of expression patterns was confirmed using semi quantitative RT-PCR and western blot analysis. In particular, the protease activated receptor-1 (PAR-1) gene that encodes a receptor for thrombin was down regulated. This is the first demonstration that PAR-1 is down regulated by the progestogen LNG in human endometrium. Alteration in the expression levels of this receptor may affect both growth and haemostatic activity within the endometrium and may account for the observed morphological effects seen in users of intrauterine LNG delivery devices. PMID- 12549866 TI - Quantitative assessment of cerebral neocapillary network and its remodeling in mice using intravital fluorescence videomicroscopy. AB - To assess the responses of different growth factors on cerebral neocapillary density (NCD), cerebral angiogenesis was induced in mice using growth factors, basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF) at a concentration of 6 ng/ml each. Intravital fluorescence videomicroscopy was used to quantitatively evaluate microhemodynamic parameters such as diameter and red cell velocity. The gel-nylon mesh-sandwich system was implanted over the exposed cortex. After incubation for different periods of time (days 7, 14 or 28), fluorescein isothiocyanate (FITC)-labeled red cells were injected through a carotid artery and the neocapillaries on the upper surface of the nylon mesh were observed under a fluorescence videomicroscope. Based on the recorded videoimages, we evaluated the density, diameter and red cell velocity of the neocapillaries. The NCD in the bFGF group on day 7 was significantly higher than that in the PDGF group on day 7 (P < 0.01). The NCD (index) reached 100% on day 14, while it reduced significantly in both the groups on day 28. The neocapillary diameter was greater than that of the pre-existing capillaries on day 7. On day 14, a clear difference appeared in the capillary density between large and small vessels. The red cell velocity increased with the number of days after incubation. The response of cerebral neocapillaries to acetylcholine was measured after 28 days of incubation with growth factor bFGF and with PDGF. The red cell velocity increased significantly from its basal value in the PDGF group. These results suggest that the neocapillaries in the PDGF group matured earlier than those in the bFGF group. PMID- 12549867 TI - Serum and plasma levels of FGF-2 and VEGF in healthy blood donors. AB - VEGF-A and FGF-2 are two angiogenic growth factors involved in the growth and invasion of solid tumours and haematological malignancies. They are also believed to play an important role in cardiovascular and inflammatory diseases. Several studies dealing with measurement of circulating FGF-2 and VEGF have been published during the last decade. We have studied the levels of FGF-2 and VEGF-A in serum and EDTA plasma from 80 healthy blood donors. The samples were analyzed using the most widely used and commercially available ELISAkits. There was no significant effect of age on any of the assays and no effect of sex on P-FGF-2, S VEGF-A and P-VEGF-A. Using the 97.5th percentile we obtained the following reference values: P-FGF-2 < 6.4 ng/l; S-VEGF-A < 500 ng/l; P-VEGF-A < 80 ng/l. Separate gender based reference intervals were made for S-FGF-2 as women had significantly higher S-FGF-2 values. Reference values for S-FGF-2 were <4.0 ng/l (men) and <10.8 ng/l (women). PMID- 12549868 TI - Effect of human recombinant Endostatin protein on human angiogenesis. AB - Tumor growth and metastasis are dependent on the development of new blood vessels. Inhibitors of new vessel growth have been widely investigated as anti tumor agents. Endostatin, a 20 kDa C-terminal fragment of collagen XVIII inhibits endothelial cell proliferation, induces endothelial cell apoptosis, and can both inhibit and reverse tumor growth in mice. However, human recombinant endostatin has had limited testing against human tissue targets. To investigate the effect of human endostatin on a human vessel target over a broad range of concentrations (10(-l2)-10(-4) M), human placental vein disks were grown for a period of 2 weeks in a 0.3% fibrin clot overlayed with growth medium. Disks from five individual placentas were tested. For each placenta utilized, a control (medium and 20% fetal bovine serum [FBS]) group and a group treated with heparin (300 microg/ml) and hydrocortisone 21-phosphate (350 microg/ml) (heparin-steroid) at a dose known to inhibit angiogenesis were included. Endostatin was tested at concentrations of 10(-12)-10(-4) M in medium containing 20% FBS. The rate of initiation and the angiogenic growth index (on a visually graded semi-quantitative scale of 0-16) were determined for all experimental conditions. Endostatin inhibited angiogenesis in our model only in high concentrations. At 10(-5) M, endostatin did not alter the percent of wells that initiated an angiogenic response, but significantly inhibited subsequent vessel growth. At 10(-4) M, endostatin was able to inhibit both initiation and subsequent new vessel growth. Human endostatin can inhibit the initiation of a human angiogenic response and inhibit the subsequent proliferation of human neovessels when used at high doses in a continuous exposure model. PMID- 12549869 TI - Coronary ischemia/reperfusion increases proliferating cell nuclear antigen in vascular endothelial cells in rat hearts. AB - To elucidate the sequence of early events in the increase in capillarity caused by reperfusion following transient coronary occlusion, the time course of expression of proliferating cell nuclear antigen (PCNA) was studied immunohistochemically in rat hearts subjected to different periods of reperfusion after a 3 minute occlusion. Twenty-one male Wistar rats were killed after different periods of reperfusion following occlusion of the coronary artery for 3 minutes. The left ventricles were removed and paired serial sections were treated immunohistochemically for PCNA or stained to show the enzymes characteristic of the arteriolar, intermediate and venular portions of the capillary bed. The time course of PCNA expression, and its distribution in relation to the different portions, was examined. An increase in PCNA-expressing nuclei was found at 24 hours after the start of reperfusion; numbers reached a maximum between 72 and 96 hours, and at 168 hours had decreased again. The majority of PCNA-expressing elements were localized to the dipeptidylpeptidase IV-reactive (i.e. venular) capillary portions, with some in the intermediate and alkaline phosphatase reactive (i.e. arteriolar) portions. The distribution of PCNA in the early reperfusion period suggests that angiogenesis after transient ischemia occurs mainly from the venular side of the capillary bed, with some contribution from intermediate and arteriolar capillary portions. PMID- 12549870 TI - Stoichiometric noncovalent interaction in molecular imprinting. AB - In this review article the function of the binding site monomers in the molecular imprinting procedure is discussed. Especially, new developments towards stoichiometric noncovalent interactions are highlighted. In stoichiometric noncovalent interactions template and binding site monomer in an 1:1 molar ratio are nearly completely bound to each other. This is only possible if the association constants are considerably high (Kass > 900 M(-1)). Using this type of interaction in molecular imprinting no excess of binding sites is necessary and binding sites are only located inside the imprinted cavity. Since all cavities can be reloaded these polymers show high capacity (e.g., for preparative application) and are especially suited for the synthesis of catalytically active imprinted polymers. Discussed are binding site interactions based on amidines (and guanidines), multiple hydrogen bonding, charge-transfer interactions, and host-guest inclusion. The systematic investigation of the underlying binding reaction is described in detail. With low-molecular weight model substances the thermodynamics of the association can be conveniently investigated, e.g., by NMR spectroscopy. PMID- 12549871 TI - Molecularly imprinted polymeric membranes. AB - Molecularly imprinted polymeric membranes have been emerged since 1990. Among various kinds of molecular imprinting studies, the application of molecular imprinting to membrane separation is still a novel investigation. In the present review paper, molecularly imprinted polymeric membranes are summarized and examined. The application of molecular imprinting to membrane separation shortly leads to high performance separation membranes. PMID- 12549872 TI - Novel MIP formats. PMID- 12549873 TI - Molecular imprinting of surfaces. AB - Recent years have seen the development of a number of novel strategies for improving the performance of molecularly imprinted polymers and for adapting them to new application areas. The imprinting of surfaces has received significant attention over the past few years, due to the possibilities available in terms of the imprinting of macromolecular structures and for avoiding problems with mass transfer. Furthermore, some surface imprinting protocols offer unique possibilities for a number of application areas. In this review we present an analysis of the approaches thusfar employed for the imprinting of surfaces and discuss the consequences of these strategies for application development. PMID- 12549874 TI - Investigation of disaccharide recognition by molecularly imprinted polymers. AB - The selectivity of carbohydrate-imprinted polymers for several disaccharides, namely cellobiose, maltose, lactose and gentiobiose, is investigated. An ternary ligand-Cu(II)-carbohydrate complex was formed in alkaline solution and captured afterwards in the polymer. The accessibility of the polymer matrix for disaccharides was investigated by HPLC analysis, refractometry and 1H NMR spectroscopy applying excess of the original template during rebinding experiments under saturation conditions in unbuffered, aqueous solution at neutral pH and 20 degrees C. The selective discrimination of the alpha- and beta glycosidic linkage of cellobiose and maltose is demonstrated. It is further shown, that the disaccharide-imprinted polymers slightly distinguish between the 1,4-beta- and the 1,6-beta-glycosidic linkage of cellobiose and gentiobiose, while cellobiose and lactose are not selectively recognized. Due to the weak apparent binding constant of the functional Cu(II) monomers with the targeted disaccharides at physiological pH, the recognition process is dominated by the shape of the created imprinted cavity under the applied conditions. PMID- 12549875 TI - Surface imprinting polymers for the recognition of nucleotides. AB - A highly selective polymer has been prepared for the selective separation of nucleotides by the surface imprinting polymerization. A dialkyl quaternary ammonium chloride was effective as the functional molecule for recognizing the difference in the structure of nucleotides. Adsorptive behavior of the ionic species of the structural analogues, inosine-5'-monophosphoric acid (IMP) and guanosine-5'-monophosphoric acid (GMP), could be controlled by changing the pH condition. Surface imprinting polymers were prepared under different pH conditions; pH 9.0 and pH 8.5. The IMP-imprinted polymers exhibited higher template effect for IMP than for a structural analogue, GMP. A reference polymer prepared without the imprint molecule neither exhibit any selectivity to IMP nor to GMP. The adsorption behavior was quantitatively evaluated by the binding constants for the IMP-imprinted polymer. The imprinting polymer was found to recognize a small structural difference in nucleotides. PMID- 12549876 TI - Relationship between enantioselectivity of alternative molecularly imprinted polymeric membranes and species of amino acid residues composing chiral recognition sites. AB - Molecularly imprinted polymeric membranes with tetrapeptide residue H-Asp(OcHex) Asp(OcHex)-Asp(OcHex)-Asp(OcHex)-CH2- (DDDD) or H-Glu(OBzl)-Glu(OBzl)-Glu(OBzl) Glu(OBzl)-CH2- (EEEE) were prepared during membrane preparation (casting) processing in the presence of print molecules. The Boc-L-Trp imprinted polymeric membranes thus obtained showed adsorption selectivity toward Ac-L-Trp from its racemic mixtures. From adsorption isotherms of Ac-Trp, the chiral recognition site, that had been formed by the presence of print molecules in the membrane preparation process, exclusively recognized Ac-L-Trp that possessed the same configuration of the print molecule. The affinity constants between chiral recognition sites in the membrane and Ac-L-Trp was determined to be 1.00 x 10(4) mol(-1) dm3 and 1.08 x 10(4) mol(-1) dm3 for the DDDD and EEEE membranes, respectively. Enantioselective electrodialysis could be attained by applying an optimum potential difference to give permselectivity, with a value close to its adsorption selectivity. PMID- 12549877 TI - Development of an aspartic acid-based cross-linking monomer for improved bioseparations. AB - Improved specificity and binding affinity by molecularly imprinted polymers is possible by development of novel functional materials. Furthermore, increasing the cross-link density of imprinted polymers by using cross-linking functional groups was anticipated to improve polymer molecular recognition. A novel cross linking monomer derived from an L-aspartic acid precursor was synthesized and employed in molecularly imprinted polymers to mimic more closely the scaffolding of proteins, and thus provide more protein-like selectivity. Chromatographic results revealed a more than 7-fold improvement in polymers imprinted using the new monomer versus a traditionally formulated polymer imprinted with methacrylic acid as the functional monomer. PMID- 12549878 TI - HPLC-based bioseparations using molecularly imprinted polymers. AB - HPLC-based separations of amino acids and peptides, nucleotide bases, drugs, sugars and steroids using molecularly imprinted polymers (MIPs) have been reviewed in this article. The molecular recognition mechanisms of the template molecules on the MIPs in organic and aqueous eluents were discussed. Furthermore, new polymerization methods suitable for preparations of HPLC columns and packing materials using molecular imprinting techniques, and their applications to HPLC based separations are also dealt with. PMID- 12549879 TI - Selective solid-phase extraction of bio- and environmental samples using molecularly imprinted polymers. AB - Of the many applications of molecular imprinting in analytical separation science, the one with highest potential of soon being used in routine analysis is that of solid-phase extraction. Already several examples of selective pre concentration of biological and environmental samples have been reported. The interest in imprinted extraction sorbents originates from the high selectivities and affinities obtainable, properties which can be qualitatively and quantitatively pre-determined for a particular analyte and separation by the imprinting process. This review summarises work published on molecular imprinted solid-phase extraction and discusses some imprinted-sorbent specific method development issues. PMID- 12549880 TI - MIP-ligand binding assays (pseudo-immunoassays). AB - Molecular imprint sorbent assays (MIAs) have been applied to an increasing number of analytes of medical and environmental interest: the sensitivities and selectivities of these assays are comparable to immunoassays employing biological antibodies. In a number of cases complete analytical procedures starting from raw samples (blood, plasma and urine) have been demonstrated. There have been significant advances in applying MIPs in new formats and in the use of non radioisotope labels. Progress in the field is reviewed, with particular emphasis on the technical aspects and new innovations. It is demonstrated that many of the perceived drawbacks of molecular imprinted polymers (MIPs) do not hinder their application in competitive binding assays: Many MIAs have been applied in aqueous systems and a heterogenous distribution of binding sites is not problematic, provided the recognition sites which bind the probe most strongly are selective. PMID- 12549881 TI - Molecular imprinting and solid phase extraction of flavonoid compounds. AB - Molecularly imprinted polymers (MIPs) for quercetin have been successfully prepared by a thermal polymerization method using 4-vinylpyridine (4-VP) and ethylene glycol dimethacrylate (EDMA) as functional monomer and cross-linker, respectively. The obtained molecularly imprinted polymers were evaluated by HPLC using organic eluents, with respect to their selective recognition properties for quercetin and related compounds of the flavonoid class. Two equivalent control polymers, a blank polymer and a polymer imprinted with a structural analogous template, were synthesized, in order to confirm the obtained results. Furthermore, preliminary experiments confirm the applicability of the prepared MIPs for solid phase extraction (SPE), as rapid and facile clean-up of wine samples for HPLC analysis is an envisaged field of application. The successful preparation of molecularly imprinted polymers for flavones provides an innovative opportunity for the development of advanced separation materials, with applications in the field of wine and fermentation analysis. PMID- 12549882 TI - A molecular imprinted polymer with recognition properties towards the carcinogenic mycotoxin ochratoxin A. AB - A molecularly imprinted polymer which recognises the mycotoxin ochratoxin A was prepared using the mimic N-(4-chloro-1-hydroxy-2-naphthoylamido)-(L) phenylalanine as a template. The polymer was obtained by dissolving the template, methacrylic acid and ethylendimethacrylate in chloroform and polymerising the mixture by thermal treatment at 60 degrees C. The monolith obtained was crushed, sieved to 30-90 microm and extensively washed till the template could no longer be found in the washing solution. The binding properties towards the template, ochratoxin A and several related molecules were measured by eluting with acetonitrile and chloroform a HPLC column packed with the imprinted polymer. The experimental results show that the polymer recognises not only the template well, but also the ochratoxin A. The specific molecular recognition effect is due to hydrogen bond interactions but in order to assure the full recognition effect adjunctive steric factors are necessary. The magnitude of these interactions can be controlled by the use of limited amounts of acetic acid in the mobile phase. From the measurement of the relative selectivity it was found that only the simultaneous presence of the carboxyl, the phenolic hydroxyl and certain peculiar substructures such as the chlorine atom assures the whole recognition of the template. PMID- 12549883 TI - Molecularly imprinted polymer as storage medium for an analyte. AB - A molecularly imprinted polymer (MIP) able to bind theophylline with a considerable degree of selectivity was prepared through a noncovalent route. The polymer after equilibrating with the template molecule (theophylline) and storage for various periods of time, the amount of theophylline desorbed from the polymer was found to be effectively quantitative indicating that MIPs could be used as a storage matrix for analytes. The stored analyte could be desorbed and analysed conveniently. PMID- 12549884 TI - Study of the factors influencing peak asymmetry on chromatography using a molecularly imprinted polymer prepared by the epitope approach. AB - Investigations of the effect of sample load on peak asymmetry during chromatography on molecularly imprinted polymer prepared by the epitope approach showed that the shape of the peaks for the template Tyr-Pro-Leu-Gly-NH2 and for acetyl-L-tyrosine ethyl ester changed considerably until a split was observed. In contrast, the asymmetry of the peaks corresponding to oxytocin, which possesses the same C-terminus tripeptide as the template and interacts with the imprinted polymer, remained essentially unaltered. The circular dichroism (CD) spectra of these peptides showed significant dependence on peptide concentration, and the dependence was nearly the same for all the tested peptides. The addition of acetic acid influenced the CD spectra of YPLG and oxytocin but had no influence on the spectrum of acetyl-L-tyrosine ethyl ester. The shape differences in the chromatographic peaks seem to be associated with a solvation mechanism rather than with solute-solute complexation in solution. However, the observed differences in peak asymmetry cannot be completely explained by the mechanisms that have been postulated previously. Our results suggest the formation of triple complexes between a solute molecule (or molecules), an already adsorbed solute molecule, and an adjacent region of the polymeric stationary phase. These triple complexes may influence the retention of analytes and contribute to peak asymmetry. PMID- 12549885 TI - The voices of qualitative researchers: sharing the conversation. PMID- 12549886 TI - Reflections on self in qualitative research: stories of family. AB - Reflexivity is a qualitative research strategy that addresses our subjectivity as researchers related to people and events that we encounter in the field. Reflexivity also addresses the subjective nature of the research account as a narrative constructed by us as researchers. Reflexivity enhances the quality of research through its ability to extend our understanding of how our positions and interests as researchers affect all stages of the research process. The reflexive account presented here frames the analysis and interpretation of previously published findings on work and play in families by highlighting aspects of the researcher's reflexivity across the entire research process, including situating the study, gaining access, managing self, living in the field, and telling the story. Its purpose is to demonstrate use of reflexivity in qualitative research as a strategy to consider our subjectivity as researchers and serve as signposts for readers about what is happening throughout the research process. PMID- 12549887 TI - Professional tensions in client-centered practice: using institutional ethnography to generate understanding and transformation. AB - For almost 20 years, occupational therapists have advocated client-centered practice. Yet client-centered practice is fraught with tensions that arise outside the practice of individual occupational therapists. This paper is guided by two questions: What produces professional tensions in client-centered practice? and What understanding and change might be generated using institutional ethnography? The sociological theory and method of institutional ethnography are described using data from an ongoing investigation of mental health services as a social institution. Illustrated are the research aim, research questions, and institutional analysis that distinguish institutional ethnography from conventional ethnography. Two professional tensions are associated with attempts to fulfill client-centered practice in mental health. One is that of working at cross-purposes with the prevailing hierarchical structure; the other tension is that of being celebrated yet subordinated in the medical and management hierarchies of health services. Although client-centered practice is difficult to do, the authors recommend institutional ethnography as a research approach to generate understanding and transformation of the context and practice of occupational therapy. PMID- 12549888 TI - Gazing anew: the shift from a clinical gaze to an ethnographic lens. AB - The purpose of this paper is to analyze ways of perceiving, knowing, and being with others while engaging in qualitative and ethnographic research. As occupational therapists acquire research expertise and embark on research trajectories, they bring their clinical legacy into the research arena. The ability to conduct qualitative and ethnographic projects may require a reconfiguration of the clinical gaze of occupational therapists. This transformation is complex and also involves acquiring and adopting a stance that alters relational and interpersonal processes and results in new ways of seeing and being. I draw upon experiences in training, mentoring, and supervising therapists who embark on ethnographic research, as well as my own research practices, to illustrate points of intersection and divergence between the clinical gaze and ethnographic lens. Artistic metaphors are also used to highlight aspects of the interactional and intersubjective processes intrinsic to ethnographic practices. I argue that ethnographers are continually evaluating and negotiating their stance in the field. The clinician who becomes an ethnographer faces unique challenges to his or her stance in the midst of moment to moment encounters as well as over time in the course of prolonged engagement. PMID- 12549890 TI - Data analysis in qualitative research: a plea for sharing the magic and the effort. AB - This discussion of data analysis in qualitative research addresses the question of how authors describe this aspect of their research. I suggest that the tradition of organization of research papers from quantitative research is not a good fit for writing qualitative research. I argue for less jargon and more detailed description, with the analytic process integrated into the findings and interpretation. PMID- 12549889 TI - The perils of power in interpretive research. AB - Occupational therapy is based on core values of altruism, equality, and honoring the dignity of others. Embedded in these values is the ever-present negotiation of power. To honor the concern for the welfare of others, researchers are challenged to think about issues of power throughout the research process. This paper identifies dilemmas and raises questions researchers might ask themselves as they struggle to share power in the interpretive research process. PMID- 12549891 TI - Qualitative data analysis: challenges and dilemmas related to theory and method. AB - The purpose of this article is to initiate a discussion of the struggles and challenges we encountered as we developed a method of analysis for a particular qualitative study. We present our thinking process showing the questions that arose, the theoretical ideas on which we relied, and the decisions we made at crucial junctures. Portions from several theoretical frames used as the basis for the analysis are presented and include Polkinghorne's (1995) narrative inquiry, Glaser and Strauss's (1967) and Charmaz's (2000) grounded theory, Riessman's (1990b) mixed approach to analysis, and Rosaldo's (1989) ideas on reflexivity. Examples from the study are used to illustrate points in the analysis. PMID- 12549892 TI - Using participant observation to study the meaning of occupations of young children with autism and other developmental disabilities. AB - Understanding the individual meaning of daily activities for children with developmental disabilities such as autism is both important and challenging for researchers and practitioners. Rigorous participant observation offers a method for developing this knowledge base by including the child's perspective. Through literature and examples from an ethnography of young children with autism, this article illustrates the application of participant observation to children with developmental disabilities. Specific strategies can promote valid interpretations despite developmental, linguistic, and perceptual differences between adult researchers and child participants. PMID- 12549893 TI - Occupational therapy and participatory research: a partnership worth pursuing. AB - This paper argues that participatory research is an approach to developing knowledge that can make significant contributions to occupational therapy. One project, working with older adults in Toronto to organize a new seniors' organization, is used to illustrate the ways in which this approach is participatory and how it can be understood as research. Other examples of participatory research from the occupational therapy literature are then presented. These include research among occupational therapists and research with participants who are typically clients of occupational therapy services. The examples are used to illustrate the conceptual links between client-centered, occupation-based occupational therapy and participatory research. Challenges of adopting a participatory research approach in occupational therapy are discussed, although many of these can be addressed through open discussions with participants. The paper concludes that participatory research is an approach to research that should be pursued more actively in occupational therapy. PMID- 12549894 TI - Encounters in a world of pain: an autoethnography. AB - Pain, a major health problem in the United States, is a highly complex and subjective experience that is poorly understood by many medical, psychological, and rehabilitation practitioners. In this paper I use a qualitative research methodology, autoethnography, to present a personal narrative about my experience of chronic pain. In this research I am both the research participant and the researcher. I begin with my personal narrative. I then problematize conceptions about chronic pain and discuss them from the point of view of my own narrative and from stories and ethnographies in the literature. Finally I reflect on how occupational therapists can more effectively work with persons with chronic pain. PMID- 12549895 TI - The effect of body orientation on a point-to-point movement in healthy elderly persons. AB - OBJECTIVE: Upper limb retraining during the early phases of neurological rehabilitation often involves having individuals practice reaching in body orientations that reduce the effect of gravity on various joint motions (e.g., shoulder flexion, elbow extension). However, the efficacy of these training techniques has not been determined. The purpose of this study was to determine the effects of reducing gravity through a change in body orientation from sitting to side-lying on the kinematics of a point-to-point movement in healthy elderly persons. METHOD: Nine healthy, right-hand-dominant women 62 to 66 years of age pointed to a target in two different body orientations-sitting and side-lying. A 2-dimensional kinematic analysis of the movement was performed to compare the trajectory of the hand and the interjoint coordination under the two conditions. RESULTS: Regardless of body orientation relative to gravity, participants produced straight hand paths and smooth, bell-shaped velocity profiles. However, they moved slower in side-lying, and the pattern of interjoint coordination varied. The shoulder and elbow moved less, whereas the scapula made a greater contribution to the overall movement. Furthermore, the temporal coordination of the joints was modified as a consequence of body position. CONCLUSION: The results indicate that point-to-point arm movements made against gravity differ from those made in a gravity-reduced plane, particularly at the joint level, illustrating that movement organization is sensitive to this contextual difference. The effect of minimizing gravity on upper limb movement needs to be explored in patient populations to determine whether training patients in gravity reduced orientations is efficacious. PMID- 12549897 TI - Where is the individual in statistics? PMID- 12549896 TI - Test-retest reliability of the Purdue Pegboard for persons with multiple sclerosis. AB - OBJECTIVE: The Purdue Pegboard test often is used in clinical settings to evaluate changes in clients' fine motor dexterity. The purpose of this study was to determine the test-retest reliability and practice effects of the Purdue Pegboard for persons with multiple sclerosis. In addition, this study compared the reliability of one-trial administration to three-trial administration of the four subtests. METHOD: Thirty-two volunteers from a midwestern community-based maintenance rehabilitation center for persons with multiple sclerosis participated in this study. The participants were administered the four subtests of the Purdue Pegboard, three trials in a row. A second administration was completed 1 week later. Data from 25 participants were analyzed using paired t tests, Pearson product-moment correlations, and intraclass correlation coefficients. RESULTS: The test-retest reliability coefficients ranged from .85 to .90 for one-trial administration and from .92 to .96 for the sum of three trials. No significant practice effects existed except for the sum of three trials of both hands. CONCLUSION: This study suggests that the one-trial administration of the Purdue Pegboard is a sufficiently reliable assessment to use with persons with multiple sclerosis. Findings further suggest that for a person with multiple sclerosis, any changes in Purdue Pegboard scores using one trial administration may reflect actual change in that person's dexterity, as no practice effect was demonstrated in this study. PMID- 12549898 TI - Pertussis outbreak among adults at an oil refinery--Illinois, August-October 2002. AB - On September 16, 2002, the Crawford County Health Department (CCHD) reported to the Illinois Department of Public Health (IDPH) four cases of cough illness among workers at an oil refinery (total worker population: 750) in Crawford County, Illinois. On August 14, a worker aged 39 years reported to the plant's health unit with a cough lasting 14 days. On the same day, the worker's supervisor aged 50 years visited the health unit for a paroxysmal cough of 3 days' duration and an incident of cough syncope. Both patients were referred to private health-care providers; blood samples from both patients had serologic test results suggestive of recent Bondetella pertusis infection, and CCHD was contacted. On September 18, IDPH and CCHD initiated active surveillance and case investigations. This report summarizes the results of that investigation, which found that during August 1 October 9, pertussis was diagnosed in 15 (10%) of 150 oil refinery workers from two separate operations (n=95) and maintenance (n=55) complexes, who were linked by contact with the ill supervisor. Through enhanced case finding, 24 cases of pertussis, 21 (88%) of which occurred in adults aged > or = 20 years, were identified in this outbreak, underscoring the need to recognize this highly infectious disease in adults and to improve national diagnostic and preventive strategies. PMID- 12549899 TI - Accelerated measles control--Cambodia, 1999-2002. AB - Cambodia is recovering from approximately 30 years of civil war that resulted in the breakdown of the country's public health infrastructure. In 1999, the Ministry of Health initiated a measles-control program with the goal of reducing the annual incidence of measles to <10,000 cases in 2005 by strengthening measles surveillance, improving routine vaccination coverage, implementing supplementary measles immunization activities (SIAs), and providing vitamin A during outbreak investigations and SIAs. This report summarizes measles-vaccination activities and their impact in reducing reported measles cases from 13,827 in 1999 to 1,234 in 2002 and suggests options for future measles-control efforts in postconflict situations. PMID- 12549900 TI - Tobacco use among middle and high school students--New Hampshire, 1995-2001. AB - Tobacco use is the leading cause of preventable death in the United States. Because 80% of adult smokers began smoking as minors, efforts to prevent smoking initiation have focused on adolescents. To examine trends in smoking prevalence among adolescents, the New Hampshire Department of Health and Human Services analyzed data from the New Hampshire Youth Risk Behavior Survey (NHYRBS) and the New Hampshire Youth Tobacco Survey (NHYTS) during 1995-2001. This report summarizes the results of that analysis, which indicate that smoking prevalence in New Hampshire has declined among both middle and high school students. When fully operational, New Hampshire's comprehensive tobacco-prevention and -control program should lead to further reductions in smoking among adolescents and begin to decrease smoking among adults. PMID- 12549901 TI - MAP kinases and CDKs: kinetic basis for catalytic activation. AB - Protein kinases constitute one of the largest enzyme families encoded by the human genome. Owing to their critical role in virtually all aspects of signal transduction, protein kinases have evolved stringent mechanisms for their regulation, which classically falls into two categories: regulation by pseudosubstrate autoinhibitory domains, and remodeling of the catalytic core in response to phosphorylation and/or protein/protein interactions. While the action of pseudosubstrate domains can be explained by simple competitive autoinhibition kinetics, it is less well understood how active site phosphorylation and/or protein/protein interactions alter rates of catalysis. Here, the kinetic basis for kinase activation is discussed in relation to the MAP kinase, ERK2, and the cyclin-dependent kinase, CDK2/cyclin A, two enzymes of central importance to mammalian cell growth and division, and which serve as prototypic models of nonautoinhibitory regulation. PMID- 12549902 TI - The immunosuppressive agent mizoribine monophosphate forms a transition state analogue complex with inosine monophosphate dehydrogenase. AB - Mizoribine monophosphate (MZP) is the active metabolite of the immunosuppressive agent mizoribine and a potent inhibitor of IMP dehydrogenase (IMPDH). This enzyme catalyzes the oxidation of IMP to XMP with the concomitant reduction of NAD via a covalent intermediate at Cys319 (E-XMP). Surprisingly, mutational analysis indicates that MZP is a transition state analogue although its structure does not resemble that of the expected transition state. Here we report the X-ray crystal structure of the E.MZP complex at 2.0 A resolution that reveals a transition state-like structure and solves the mechanistic puzzle of the IMPDH reaction. The protein assumes a new conformation where a flap folds into the NAD site and MZP, Cys319, and a water molecule are arranged in a geometry resembling the transition state. The water appears to be activated by interactions with a conserved Arg418 Tyr419 dyad. Mutagenesis experiments confirm that this new closed conformation is required for the hydrolysis of E-XMP, but not for the reduction of NAD. The closed conformation provides a structural explanation for the differences in drug selectivity and catalytic efficiency of IMPDH isozymes. PMID- 12549903 TI - Tautomeric rearrangement of a dihydroflavin bound to monomeric sarcosine oxidase or N-methyltryptophan oxidase. AB - Monomeric sarcosine oxidase (MSOX) and N-methyltryptophan oxidase (MTOX) are homologous bacterial flavoenzymes that contain covalently bound flavin [8alpha-(S cysteinyl)FAD]. Reaction of MSOX or MTOX with a small excess of sodium borohydride results in immediate flavin reduction to a species that exhibits spectral properties (lambda(max) = 405 nm with a second broad peak at 332 nm) similar to those of 3,4-dihydroflavin. The borohydride-reduced enzymes retain full catalytic activity. Substrate reduction converts the 405 nm species to an air-sensitive tetrahydroflavin that reacts with oxygen to yield unmodified oxidized enzyme. Unexpectedly, the putative 3,4-dihydroflavin bound to MSOX or MTOX is unstable in the absence of substrate. An isosbestic conversion of the 405 nm species to yield unmodified, oxidized flavin is observed when the reaction is conducted under aerobic conditions (k(obs) = 4.9 x 10(-2) min(-1)). Under anaerobic conditions, an oxygen-sensitive species resembling 1,5-dihydroflavin is formed in an isosbestic reaction that occurs at a rate similar to that of the aerobic reaction (k(obs) = 5.3 x 10(-2) min(-1)). Possible reaction of the 3,4 dihydroflavin with a second molecule of borohydride to yield an air-sensitive tetrahydroflavin is unlikely since prior scavenging of residual borohydride with excess formaldehyde had no effect on the aerobic conversion to unmodified oxidized flavin. The observed instability is attributed to a tautomeric rearrangement of the 3,4-dihydroflavin to generate 1,5-dihydroflavin, a species that is also air-sensitive. Evidence in favor of an active site facilitated tautomerization reaction is provided by the fact that the stability of the 405 nm species formed with MSOX is enhanced 200-fold upon denaturation with urea or heat. The observed tautomeric rearrangement of 3,4-dihydroflavin may provide insight regarding a related flavin tautomerization reaction that has been proposed as a key step in the biosynthesis of covalent flavin linkages. PMID- 12549904 TI - Mutational analysis of roles for extracellular cysteine residues in the assembly and function of human alpha 7-nicotinic acetylcholine receptors. AB - Nicotinic acetylcholine receptors (nAChR) containing alpha7 subunits self assemble into simple, homopentameric complexes. However, successful heterologous expression of functional alpha7-nAChR has only been achieved in a few host cell types, such as the SH-EP1 human epithelial cell line. All ionotropic glycine receptor, GABA(A) receptor, 5-HT(3) receptor, and nAChR subunits contain a pair of highly conserved cysteine residues (C150 and C164 for alpha7 subunits) in their N-terminal extracellular domain. These residues are thought to be involved in the formation of a conserved cystine loop that is critical to the proper folding and assembly of subunits. However, nAChR alpha7 (and alpha8) subunits also contain a third cysteine residue, C138, N-terminal to the conserved cysteine pair. Using SH-EP1 cells as a host for heterologous expression, we evaluated the roles of C138, C150, and C164 in subunit folding, assembly, and cell surface expression and function of alpha7-nAChR. Results indicate that mutation of C138, but not of C150 or C164, yields an nAChR that can assemble to form (125)I-labeled alpha-bungarotoxin binding sites expressed on the cell surface. Further, whole cell patch clamp recordings demonstrate that mutation of C138 to alanine does not alter the function of the fully assembled alpha7-nAChR. These results indicate that C150 and C164 are required for surface expression, but that C138 is neither necessary for nor inhibitory toward the surface expression and function of human alpha7-nAChR. These results suggest that disulfide bond formation between C138 and either C150 or C164, if it occurs, has no significant effect on alpha7-nAChR assembly or function. PMID- 12549905 TI - Monitoring expression and clustering of the ionotropic 5HT3 receptor in plasma membranes of live biological cells. AB - The ionotropic 5HT(3) receptor was expressed in transiently transfected mammalian cells, yielding an unprecedented high concentration of up to 12 million receptors per cell. Receptor traffic in the plasma membrane of live cells was observed continuously over 24 h by fluorescence scanning confocal microscopy. This was possible by using 5HT(3) receptor-specific fluorescent ligands with high binding affinity and low off-rate to pulse label receptors at any time after appearance on the cell surface, and label subsequently those receptors expressed later by another, spectrally distinguishable, high-affinity fluorescent ligand. Having reached a critical cell surface concentration of approximately 3000 receptors/microm(2), the receptors started to aggregate in patches with a 4-fold increased surface concentration. The clusters were constantly delivered from a pool of freshly expressed receptors isotropically distributed within the basolateral region of the cell membrane. From there, they migrated to and accumulated on the apical cell surface approximately 9 h after transfection. Individual clusters grew until they reached a critical size of 1-2 microm when they merged to form with 3-5 microm large macroclusters. Clustered receptors were immobile on the minute time scale but always coexisted with monomeric receptors in the regions surrounding the clusters as revealed by fluorescence correlation spectroscopy. Because the receptor density of 12 000 receptors/microm(2) in the patches is as high as that found in two-dimensional crystals of certain membrane proteins, such patches might be a proper source for direct crystallization of membrane proteins without prior purification. PMID- 12549906 TI - Structural and biochemical studies of inhibitor binding to human cytomegalovirus protease. AB - Herpesvirus protease is required for the life cycle of the virus and is an attractive target for the design and development of new anti-herpes agents. The protease belongs to a new class of serine proteases, with a novel backbone fold and a unique Ser-His-His catalytic triad. Here we report the crystal structures of human cytomegalovirus protease in complex with two peptidomimetic inhibitors. The structures reveal a new hydrogen-bonding interaction between the main chain carbonyl of the P(5) residue and the main chain amide of amino acid 137 of the protease, which is important for the binding affinity of the inhibitor. Conformational flexibility was observed in the S(3) pocket of the enzyme, and this is supported by our characterization of several mutants in this pocket. One of the structures is at 2.5 A resolution, allowing us for the first time to locate ordered solvent molecules in the inhibitor complex. The presence of two solvent molecules in the active site may have implications for the design of new inhibitors against this enzyme. Favorable and stereospecific interactions have been established in the S(1)' pocket for one of these inhibitors. PMID- 12549907 TI - Potent and selective inhibition of membrane-type serine protease 1 by human single-chain antibodies. AB - Specific human antibodies targeting proteases expressed on cancer cells can be valuable reagents for diagnosis, prognosis, and therapy of cancer. To this end, a phage-displayed antibody library was screened against a cancer-associated serine protease, MT-SP1. A protein inhibitor of serine proteases that binds to a defined surface of MT-SP1 was used in an affinity-based washing procedure. Six antibodies were selected on the basis of their ELISA profiles and ability to serve as useful immunological reagents. The apparent K(i), indicative of the potency of the antibodies at inhibiting human MT-SP1 activity, ranged from 50 pM to 129 nM. Two of the antibodies had approximately 800-fold and 1500-fold selectivity when tested against the most homologous serine protease family member, mouse MT-SP1, that exhibits 86.6% sequence identity. Surface plasmon resonance was used as an independent means of determining the binding constants of the six antibodies. Association rates were as high as 1.15 x 10(7) s(-)(1) M(-)(1), and dissociation rates were as low as 3.8 x 10(-)(4) s(-)(1). One antibody was shown to detect denatured MT-SP1 with no cross reactivity to other family members in HeLa or PC3 cells. Another antibody recognized the enzyme in human prostate tissue samples for immunohistochemistry analysis. The mode of binding among the six antibodies and the protease was analyzed by competition ELISA using three distinctly different inhibitors that mapped the enzyme surface. These antibodies constitute a new class of highly selective protease inhibitors that can be used to dissect the biological roles of proteolytic enzymes as well as to develop diagnostic and therapeutic reagents. PMID- 12549908 TI - Differential helix stabilities and sites pre-organized for tertiary interactions revealed by monitoring local nucleotide flexibility in the bI5 group I intron RNA. AB - The local environment at adenosine residues in the bI5 group I intron RNA was monitored as a function of Mg(2+) using both the traditional method of dimethyl sulfate (DMS) N1 methylation and a new approach, selective acylation of 2'-amine substituted nucleotides. These probes yield complementary structural information because N1 methylation reports accessibility at the base pairing face, whereas 2' amine acylation scores overall residue flexibility. 2'-Amine acylation robustly detects RNA secondary structure and is sensitive to higher order interactions not monitored by DMS. Disruption of RNA structure due to the 2'-amine substitution is rare and can be compensated by stabilizing folding conditions. Peripheral helices that do not interact with other parts of the RNA are more stable than both base paired helices and tertiary interactions in the conserved catalytic core. The equilibrium state of the bI5 intron RNA, prior to assembly with its protein cofactor, thus features a relatively loosely packed core anchored by more stable external stem-loop structures. Adenosine residues in J4/5 and P9.0 form structures in which the nucleotide is constrained but the N1 position is accessible, consistent with pre-organization to form long-range interactions with the 5' and 3' splice sites. PMID- 12549909 TI - Molecular recognition of the human coactivator CBP by the HIV-1 transcriptional activator Tat. AB - HIV-1 Tat is required for the expression of the viral genome. Tat binds to an RNA stem-loop and mediates the recruitment of human coactivators to facilitate HIV-1 transcription. The coactivator and acetyltransferase CREB binding protein (CBP), and the paralog p300, are recruited to the HIV-1 promoter by Tat. Here we identify the interacting domains of Tat and CBP. Circular dichroism and pulldown assays show that full-length Tat binds to the KIX domain of CBP, but not to the C/H1 or CR2 domains. Circular dichroism and NMR studies of Tat deletion mutants localize the KIX-binding domain of Tat to the N-terminal 24 residues of Tat. Transient cotransfections demonstrate that exogenous KIX behaves as a dominant negative to Tat-mediated transcription in human T-cells, suggesting that Tat and KIX interact in vivo. These findings indicate that Tat targets the KIX domain of CBP and provide insight into the molecular interactions involved in regulating HIV-1 gene expression. PMID- 12549910 TI - Residues 207, 216, and 221 and the catalytic activity of mGSTA1-1 and mGSTA2-2 toward benzo[a]pyrene-(7R,8S)-diol-(9S,10R)-epoxide. AB - Murine class alpha glutathione S-transferase subunit types A2 (mGSTA2-2) and A1 (mGSTA1-1) have high catalytic efficiency for glutathione (GSH) conjugation of the ultimate carcinogenic metabolite of benzo[a]pyrene, (+)-anti-7,8-dihydroxy 9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene, [(+)-anti-BPDE]. Only 10 residues differ between the sequences of mGSTA1-1 and 2-2. However, the catalytic efficiency of mGSTA1-1 for GSH conjugation of (+)-anti-BPDE is >3-fold higher as compared with mGSTA2-2. The crystal structure of mGSTA1-1 in complex with the GSH conjugate of (+)-anti-7,8-dihydroxy-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene (GSBpd) reveals that R216 and I221 in the last helix play important roles in catalysis [Gu, Y., Singh, S. V., and Ji, X. (2000) Biochemistry 39, 12552-12557]. The crystal structure of mGSTA2-2 in complex with GSBpd has been determined, which reveals a different binding mode of GSBpd. Comparison of the two structures suggests that residues 207 and 221 are responsible for the different binding mode of GSBpd and therefore contribute to the distinct catalytic efficiency of the two isozymes. PMID- 12549911 TI - The fusion core complex of the peste des petits ruminants virus is a six-helix bundle assembly. AB - We describe the properties of the two heptad repeats (HR1 and HR2) of the Peste des petits ruminants virus (PPRV) fusion protein (F) to obtain insights into the mechanism by which these repeats influence PPRV-mediated cell fusion. Both HR1 and HR2 inhibit PPRV-mediated syncytia formation in Vero cells in vitro. Of these, HR2 was found to be more effective than HR1. We studied the mechanism of fusion inhibition by these two repeats by using various biophysical and biochemical methods either separately or together. CD spectral analysis of these repeats revealed that the alpha-helical content of HR1 and HR2 when used together is higher than that of their simulated spectrum in the mixture, suggesting the formation of a highly structured complex by these repeats. Protease protection assays confirmed that such a complex is highly stable. Electrospray mass spectrometry of protease-digested products of the HR1-HR2 complex showed protection of fragments corresponding to both HR1 and HR2 sequences involved in complex formation. By employing size-exclusion chromatography and chemical cross linking experiments, we show that three units each of HR1 and HR2 form a complex in which HR1 is a trimer and HR2 is a monomer. Homology-based three-dimensional modeling of this complex showed that HR1 and HR2 together form a six-helix and trimeric coiled-coil bundle. In this model, the HR1 trimer forms the core whereas HR2, while interacting with HR1 in an antiparallel orientation, forms a two stranded coiled-coil structure and lies at the periphery of the structure. These results are discussed in the context of a common fusion mechanism among paramyxoviruses. PMID- 12549912 TI - Mitochondrial methionyl-tRNAfMet formyltransferase from Saccharomyces cerevisiae: gene disruption and tRNA substrate specificity. AB - Initiation of protein synthesis in bacteria, mitochondria, and chloroplasts involves a formylated methionyl-tRNA species. Formylation of this tRNA is catalyzed by a methionyl-tRNA(f)(Met) formyltransferase (formylase). Upon inactivation of the gene encoding formylase, the growth rate of Escherichia coli is severely decreased. This behavior underlines the importance of formylation to give tRNA(Met) an initiator identity. Surprisingly, however, recent data [Li, Y., Holmes, W. B., Appling, D. R., and RajBhandary, U. L. (2000) J. Bacteriol. 182, 2886-2892] showed that the respiratory growth of Saccharomyces cerevisiaewas not sensitive to deprivation of the mitochondrial formylase. In the present study, we report conditions of temperature or of growth medium composition in which inactivation of the formylase gene indeed impairs the growth of a S. cerevisiae haploid strain. Therefore, some selective advantage can eventually be associated to the existence of a formylating activity in the fungal mitochondrion under severe growth conditions. Finally, the specificity toward tRNA of S. cerevisiae mitochondrial formylase was studied using E. coli initiator tRNA and mutants derived from it. Like its bacterial counterpart, this formylase recognizes nucleotidic features in the acceptor stem of mitochondrial initiator tRNA. This behavior markedly distinguishes the mitochondrial formylase of yeast from that of animals. Indeed, it was shown that bovine mitochondrial formylase mainly recognizes the side chain of the esterified methionine plus a purine-pyrimidine base pair in the D-stem of tRNA [Takeuchi, N., Vial, L., Panvert, M., Schmitt, E., Watanabe, K., Mechulam, Y., and Blanquet, S. (2001) J. Biol. Chem. 276, 20064 20068]. Distinct tRNA recognition mechanisms adopted by the formylases of prokaryotic, fungal, or mammalian origins are likely to reflect coevolution of these enzymes with their tRNA substrate. Each mechanism appears well suited to an efficient selection of the substrate within the pool of all tRNAs. PMID- 12549913 TI - Protein and DNA residue orientations in the filamentous virus Pf1 determined by polarized Raman and polarized FTIR spectroscopy. AB - The Pseudomonas bacteriophage Pf1 is a long ( approximately 2000 nm) and thin ( approximately 6.5 nm) filament consisting of a covalently closed, single-stranded DNA genome of 7349 nucleotides coated by 7350 copies of a 46-residue alpha helical subunit. The coat subunits are arranged as a superhelix of C(1)()S(5.4)() symmetry (class II). Polarized Raman and polarized FTIR spectroscopy of oriented Pf1 fibers show that the packaged single-stranded DNA genome is ordered specifically with respect to the capsid superhelix. Bases are nonrandomly arranged along the capsid interior, deoxynucleosides are uniformly in the C2' endo/anti conformation, and the average DNA phosphodioxy group (PO(2)(-)) is oriented so that the line connecting the oxygen atoms (O.O) forms an angle of 71 degrees +/- 5 degrees with the virion axis. Raman and infrared amide band polarizations show that the subunit alpha-helix axis is inclined at an average angle of 16 degrees +/- 4 degrees with respect to the virion axis. The alpha helical symmetry of the capsid subunit is remarkably rigorous, resulting in splitting of Raman-active helix vibrational modes at 351, 445 and 1026 cm(-)(1) into apparent A-type and E(2)()-type symmetry pairs. The subunit tyrosines (Tyr 25 and Tyr 40) are oriented with phenoxyl rings packed relatively close to parallel to the virion axis. The Tyr 25 and Tyr 40 orientations of Pf1 are surprisingly close to those observed for Tyr 21 and Tyr 24 of the Ff virion (C(5)()S(2)() symmetry, class I), suggesting a preferred tyrosyl side chain conformation in packed alpha-helical subunits, irrespective of capsid symmetry. The polarized Raman spectra also provide information on the orientations of subunit alanine, valine, leucine and isoleucine side chains of the Pf1 virion. PMID- 12549914 TI - SRp55 is a regulator of calcitonin/CGRP alternative RNA splicing. AB - Alternative splicing is an important mechanism for the regulation of gene expression. The mammalian calcitonin/calcitonin gene-related peptide (CGRP) pre mRNA is alternatively spliced in a tissue-specific manner, leading to the production of calcitonin mRNA containing exons 1-4 in thyroid C cells and CGRP mRNA containing exons 1-3, 5, and 6 in neurons. The calcitonin-specific fourth exon contains an exonic splice enhancer (ESE) that binds SRp55. We define the RNA binding site of SRp55 in the ESE and demonstrate that base changes that decrease the level of SRp55 binding decrease the level of calcitonin splicing in vitro and calcitonin mRNA production in vivo. Base changes that increase the affinity of SRp55 for the ESE increase the level of calcitonin splicing in vitro and calcitonin mRNA levels in 293 cells. We also observe that SRp55 levels in different cell types correlate with the levels of calcitonin mRNA produced in these cells. Finally, we show that increasing the level of cellular expression of SRp55 stimulates calcitonin mRNA production in vivo. These observations suggest that SRp55 binding to a suboptimal RNA binding site in the calcitonin/CGRP pre mRNA ESE is required for calcitonin mRNA production. Differential amounts of SRp55 present in different cell types would then control calcitonin/CGRP alternative splicing. PMID- 12549915 TI - The pathogenic A3243G mutation in human mitochondrial tRNALeu(UUR) decreases the efficiency of aminoacylation. AB - Mutations of mtDNA, particularly those in mtDNA-encoded tRNA genes, are emerging as a significant cause of human disease. We examined the effects of the pathogenic A3243G and T3271C mutations in the mitochondrial tRNA(Leu(UUR)) gene on the aminoacylation of tRNA(Leu(UUR)). Transmitochondrial cells carrying these mutations have decreased steady-state levels of mitochondrial tRNA(Leu(UUR)). The A3243G mutation also results in a decrease in the fraction of aminoacylated tRNA(Leu(UUR)). To determine if the decreased fraction of aminoacylated tRNA(Leu(UUR)) in A3243G mutant cells was due to a defect in the ability of mutant tRNA to be aminoacylated by the human mitochondrial leucyl-tRNA synthetase, we examined the aminoacylation kinetics of wild-type and mutant tRNA(Leu(UUR)), using both native and in vitro transcribed tRNA(Leu(UUR)). Native A3243G mutant tRNA(Leu(UUR)) was 25-fold less efficiently aminoacylated in vitro, compared to native wild-type tRNA(Leu(UUR)). The T3271C mutation in tRNA(Leu(UUR)) did not affect the efficiency of aminoacylation of the native tRNA. There were no differences in aminoacylation efficiencies among wild-type and mutant tRNA(Leu(UUR)) transcripts. The combined effects of the reductions in the steady-state levels and the aminoacylated fraction of tRNA(Leu(UUR)) are likely to contribute to the decreases in the rates of mitochondrial translation observed in mutant cells. These results also suggest that the A3243G and T3271C mutations may have distinct mechanisms of pathogenesis. PMID- 12549916 TI - Heterologous production of Halorhodospira halophila holo-photoactive yellow protein through tandem expression of the postulated biosynthetic genes. AB - The photoactive yellow protein (PYP) is a bacterial photoreceptor which is the structural prototype for the PAS domain superfamily of regulators and receptors. PYP is known to have a unique p-hydroxycinnamic acid chromophore, covalently attached to a cysteine. To date, it has not been shown how holo-PYP is formed in vivo. Two genes, nearby pyp, were postulated to encode the biosynthetic enzymes, but only one was previously isolated and shown to have the requisite activity. By using a dual plasmid system, one expressing the PYP from Halorhodospira halophila and the other expressing a two-gene operon, consisting of tyrosine ammonia lyase and p-hydroxycinnamic acid ligase, we are able to present evidence that a functionally active holo-PYP can be synthesized in Escherichia coli. Plasmids containing only one of the two enzymes failed to produce holoprotein. Thus, the two genes have been shown to be both necessary and sufficient for production of holoprotein, although the activating group remains unknown. This expression system not only holds great potential for mutagenesis studies but also opens new possibilities in the search for (a) signaling partner(s) of the PYP. PMID- 12549917 TI - Effects of retinoid ligands on RIP140: molecular interaction with retinoid receptors and biological activity. AB - Receptor interacting protein 140 (RIP140) interacts with retinoic acid receptor (RAR) and retinoid X receptor (RXR) constitutively, but hormone binding enhances this interaction. The ligand-independent interaction is mediated by the amino and central regions of RIP140 which contain a total of nine copies of the LXXLL motif, whereas the agonist-induced interaction is mediated by its carboxyl terminus which contains a novel motif (1063-1076, LTKTNPILYYMLQK). The ligand independent interaction could be enhanced slightly by agonists, whereas the ligand-dependent interaction was strictly agonist dependent for both RAR and RXR. In the context of heterodimers, ligand occupancy of RXR played a more dominant role for both molecular interaction and biological activity of RIP140. Competition and mutation studies demonstrated an essential role for (1067)Asn and (1073)Met for a ligand-dependent interaction. A model was proposed to address the constitutive and agonist-dependent interaction of RIP140 with RAR/RXR. PMID- 12549918 TI - Effects of zinc occupancy on human O6-alkylguanine-DNA alkyltransferase. AB - A recent crystallographic study of recombinant human O(6)-alkylguanine-DNA alkyltransferase (hAGT) revealed a previously unknown zinc atom [Daniels et al., (2000) EMBO J. 19, 1719-1730]. The effects of zinc on the properties of hAGT are reported here. In bacterial expression systems, recombinant hAGT was produced in increasingly larger quantities when growth media are supplemented with up to 0.1 mM ZnCl(2). Metal-enriched hAGT samples had a 5-fold increase in repair rate constant over conventionally purified protein samples and a 60-fold increase over metal-stripped hAGT. In addition, mutants of the zinc-binding residues had decreases in zinc occupancy that correlated with reductions in repair rate. Zinc modulation did not abolish the repair capacity of a fraction of the hAGT population, as evidenced by the stoichiometric reaction with an oligodeoxyribonucleotide substrate. Zinc occupancy had a similar effect on the rate of reaction with O(6)-benzylguanine, a free base substrate, as on the repair of methylated DNA. Differentially zinc-treated hAGTs showed the same affinity for binding to native DNA and substrate oligodeoxyribonucleotides. Metal content manipulations had little effect upon the CD spectrum of hAGT, but fluorescence studies revealed a small conformational change based upon metal binding, and zinc occupancy correlated with enhanced hAGT stability as evidenced by resistance to the denaturing effects of urea. These results indicate that the presence of zinc confers a mechanistic enhancement to repair activity that does not result from an increase in substrate binding affinity. Zinc also provides conformational stability to hAGT that may influence its regulation. PMID- 12549919 TI - Measurement of the electronic properties of the flavoprotein old yellow enzyme (OYE) and the OYE:p-Cl phenol charge-transfer complex using Stark spectroscopy. AB - Low-temperature absorption and Stark spectroscopy have been used to study the electronic properties of oxidized flavin mononucleotide (FMN) in old yellow enzyme (OYE) and OYE complexed with p-chlorophenol (p-Cl phenol). The low temperature absorbance spectrum of OYE showed splittings of the blue and near-UV vibronic bands, which appears to be due to hydrogen bonding between the isoalloxazine moiety and the protein. A Stark spectroscopic analysis showed that the electronic structure of the FMN cofactor in OYE is not significantly perturbed relative to flavins in simple solvents. However, the charge-transfer band in the OYE:p-Cl phenol complex showed a large Stark effect indicative of substantial charge displacement. The magnitude and direction of this charge displacement are consistent with significant charge transfer along the charge transfer transition dipole moment direction. In addition, the Stark spectrum of the CT band showed unexpected fine structure that could correlate with vibrational progressions in either the p-Cl phenol donor or the flavin acceptor. PMID- 12549920 TI - Structural analysis of the PsbQ protein of photosystem II by Fourier transform infrared and circular dichroic spectroscopy and by bioinformatic methods. AB - The structure of PsbQ, one of the three main extrinsic proteins associated with the oxygen-evolving complex (OEC) of higher plants and green algae, is examined by Fourier transform infrared (FTIR) and circular dichroic (CD) spectroscopy and by computational structural prediction methods. This protein, together with two other lumenally bound extrinsic proteins, PsbO and PsbP, is essential for the stability and full activity of the OEC in plants. The FTIR spectra obtained in both H(2)O and D(2)O suggest a mainly alpha-helix structure on the basis of the relative areas of the constituents of the amide I and I' bands. The FTIR quantitative analyses indicate that PsbQ contains about 53% alpha-helix, 7% turns, 14% nonordered structure, and 24% beta-strand plus other beta-type extended structures. CD analyses indicate that PsbQ is a mainly alpha-helix protein (about 64%), presenting a small percentage assigned to beta-strand ( approximately 7%) and a larger amount assigned to turns and nonregular structures ( approximately 29%). Independent of the spectroscopic analyses, computational methods for protein structure prediction of PsbQ were utilized. First, a multiple alignment of 12 sequences of PsbQ was obtained after an extensive search in the public databases for protein and EST sequences. Based on this alignment, computational prediction of the secondary structure and the solvent accessibility suggest the presence of two different structural domains in PsbQ: a major C terminal domain containing four alpha-helices and a minor N-terminal domain with a poorly defined secondary structure enriched in proline and glycine residues. The search for PsbQ analogues by fold recognition methods, not based on the secondary structure, also indicates that PsbQ is a four alpha-helix protein, most probably folding as an up-down bundle. The results obtained by both the spectroscopic and computational methods are in agreement, all indicating that PsbQ is mainly an alpha protein, and show the value of using both methodologies for protein structure investigation. PMID- 12549921 TI - Oxidation of the two beta-carotene molecules in the photosystem II reaction center. AB - We present a spectroscopic characterization of the two nonequivalent beta carotene molecules in the photosystem II reaction center. Their electronic and vibrational properties exhibit significant differences, reflecting a somewhat different configuration for these two cofactors. Both carotenoid molecules are redox-active and can be oxidized by illumination of the reaction centers in the presence of an electron acceptor. The radical cation species show similar differences in their spectroscopic properties. The results are discussed in terms of the structure and unusual function of these carotenoids. In addition, the attribution of resonance Raman spectra of photosystem II preparations excited in the range 800-900 nm is discussed. Although contributions of chlorophyll cations cannot be formally ruled out, our results demonstrate that these spectra mainly arise from the cation radical species of the two carotenoids present in photosystem II reaction centers. PMID- 12549923 TI - Binding of Zn-chlorin to a synthetic four-helix bundle peptide through histidine ligation. AB - We have used a two histidine-containing synthetic peptide (Sharp et al. (1998) Proc. Natl. Acad. Sci. U.S.A. 95, 10465-10470) as a scaffold to bind Zn(II) chlorin e6 (ZnCe6) through histidine ligation. Protocols for the preparation and purification of the peptide using an Escherichia coli expression system are presented. Size-exclusion chromatography and circular dichroism measurements indicate that the peptide self-assembles into a four-helix bundle protein. Two variants of the peptide lacking either one or both of the histidine residues were used to demonstrate the stoichiometry of ZnCe6 binding. Comparison of the titration profiles determined by UV-vis spectroscopy for the purified one- and two-histidine peptides suggests that the two-histidine peptide can bind two ZnCe6. The binding stoichiometry of ZnCe6 was verified by gel chromatography and native gel electrophoresis using the peptide variant lacking histidine residues as the control. Like many other chlorophyll analogue molecules, ZnCe6 can be photooxidized. The light-induced electron transfer between the ZnCe6-peptide complex and the added phenyl-p-benzoquinone was measured using time-resolved EPR spectroscopy and shown to be faster and have a higher yield than the electron transfer between unbound ZnCe6 and quinone. The implications of constructing a ZnCe6-peptide complex in terms of artificial photosynthesis are discussed. PMID- 12549922 TI - Nitric oxide-induced formation of the S-2 state in the oxygen-evolving complex of photosystem II from Synechococcus elongatus. AB - In spinach photosystem II (PSII) membranes, the tetranuclear manganese cluster of the oxygen-evolving complex (OEC) can be reduced by incubation with nitric oxide at -30 degrees C to a state which is characterized by an Mn(2)(II, III) EPR multiline signal [Sarrou, J., Ioannidis, N., Deligiannakis, Y., and Petrouleas, V. (1998) Biochemistry 37, 3581-3587]. This state was recently assigned to the S( )(2) state of the OEC [Schansker, G., Goussias, C., Petrouleas, V., and Rutherford, A. W. (2002) Biochemistry 41, 3057-3064]. On the basis of EPR spectroscopy and flash-induced oxygen evolution patterns, we show that a similar reduction process takes place in PSII samples of the thermophilic cyanobacterium Synechococcus elongatus at both -30 and 0 degrees C. An EPR multiline signal, very similar but not identical to that of the S(-)(2) state in spinach, was obtained with monomeric and dimeric PSII core complexes from S. elongatus only after incubation at -30 degrees C. The assignment of this EPR multiline signal to the S(-)(2) state is corroborated by measurements of flash-induced oxygen evolution patterns and detailed fits using extended Kok models. The small reproducible shifts of several low-field peak positions of the S(-)(2) EPR multiline signal in S. elongatus compared to spinach suggest that slight differences in the coordination geometry and/or the ligands of the manganese cluster exist between thermophilic cyanobacteria and higher plants. PMID- 12549924 TI - Site-directed photochemical coupling of cytochrome b6f-associated chlorophyll. AB - Cytochrome b(6)f complexes contain a molecule of chlorophyll a (Chla), which, in Chlamydomonas reinhardtii, can be exchanged for extraneous chlorophyll during protracted incubation of the purified complex in detergent solution. The specificity of the site and its location in the complex have been studied by photochemical coupling and circular dichroism spectroscopy. Following substitution of the original chlorophyll with [(3)H]Chla, the complex was irradiated in the Soret absorption band of Chla to complete bleaching and the amount of radioactivity covalently bound to each b(6)f subunit determined. Strong labeling was found to be associated with cytochrome f. The labeling originates from [(3)H]Chla molecules bound to a slowly exchanging site and showing the properties of the endogenous Chl, not from molecules dissolved in the detergent belt surrounding the complex. Chlorophyll b (Chlb) can compete with Chla, albeit with a lower affinity. Irradiation of [(3)H]Chlb introduced into the slowly exchanging site yielded the same labeling pattern that was observed with [(3)H]Chla. Proteolytic cleavage showed [(3)H]Chla labeling to be strictly restricted to the C-terminal region of cytochrome f. Circular dichroism spectra of the native complex revealed a bilobed signal characteristic of excitonic interaction between chlorophylls. The structural and evolutionary implications of these findings are discussed. PMID- 12549926 TI - Two of the five zinc fingers in the Zap1 transcription factor DNA binding domain dominate site-specific DNA binding. AB - The Zap1 transcriptional activator from Saccharomyces cerevisiae induces expression of a series of genes containing an 11 base pair conserved promoter element (ZRE) under conditions of zinc deficiency. This work shows that Zap1 uses four of its seven zinc finger domains to contact the ZRE and that two of these dominate the interaction by contacting the essential ACC-GGT ends. Two Zn finger domains (ZF1 and ZF2) do not contact DNA, and a third ZF3 may be more important for interfinger protein-protein interactions. Zn finger domains important for ZRE contact were identified from triple mutations in Zap1, changing three residues in the alpha helix in each finger known to be important for DNA contacts in Zn finger proteins. Replacement of -1, 3, and 6 helix residues in ZF4 and ZF7 reduced the affinity of Zap1 for the wild-type ZRE. In contrast, triple mutations within the intervening ZF5 and ZF6 domains had minimal effect. The data argue that fingers 4 and 7 contact the ACC-GGT ends while fingers 5 and 6 contact the 5 bp central ZRE sequence. This conclusion is corroborated by decreased Zap1 affinity for a ZRE DNA duplex containing mutations of the AC-GT ends of the ZRE, whereas transversion mutations within the central 5 bp of the ZRE had minimal effect on Zap1 binding affinity. PMID- 12549925 TI - Distinct mechanisms by mutant presenilin 1 and 2 leading to increased intracellular levels of amyloid beta-protein 42 in Chinese hamster ovary cells. AB - To characterize the properties of presenilin (PS) 1- and PS2-associated gamma secretases, we established stable transfectants overexpressing amyloid precursor protein and wild-type (wt) or a number of mutant (mt) PS1 or PS2. Quantification of the intracellular amyloid beta-protein (Abeta) levels in mtPS1 and mtPS2 cell lines revealed the presence of two subtypes. One group consists of N141I, M239V, and T122P mutations of the PS2 gene and homologous mutations of PS1, N135D and M233T. These mutations led to an increase in the intracellular Abeta42 levels and a concomitant decrease in the intracellular Abeta40 levels. A cell-free assay for Abeta production using the membranes prepared from these transfectants exhibited predominant cleavage at position Abeta42 with marginal production of Abeta40. The other group consists of M146L, H163R, and G384A mutations of PS1, leading only to an increase in the intracellular Abeta42 levels. While the intracellular Abeta levels in M146L cells were consistent with the results from cell-free Abeta production, H163R and G384A cells showed significant discrepancies between the intracellular Abeta levels and cell-free Abeta production. Thus, all the mtPS1/2 examined here result in increases in the intracellular Abeta42 levels. This suggests that the underlying mechanisms for this shared phenotype may be diverse. PMID- 12549927 TI - Genotoxicity and mutagenicity of chromium(VI)/ascorbate-generated DNA adducts in human and bacterial cells. AB - Reduction of carcinogenic Cr(VI) by vitamin C generates ascorbate-Cr(III)-DNA cross-links, binary Cr(III)-DNA adducts, and can potentially cause oxidative DNA damage by intermediate reaction products. Here, we examined the mutational spectrum and the importance of different forms of DNA damage in genotoxicity and mutagenicity of Cr(VI) activated by physiological concentrations of ascorbate. Reduction of Cr(VI) led to a dose-dependent formation of both mutagenic and replication-blocking DNA lesions as detected by propagation of the pSP189 plasmids in human fibroblasts. Disruption of Cr-DNA binding abolished mutagenic responses and normalized the yield of replicated plasmids, indicating that Cr-DNA adducts were responsible for both mutagenicity and genotoxicity of Cr(VI). The absence of DNA breaks and abasic sites confirmed the lack of a significant production of hydroxyl radicals and Cr(V)-peroxo complexes in Cr(VI)-ascorbate reactions. Ascorbate-Cr(III)-DNA cross-links were much more mutagenic than smaller Cr(III)-DNA adducts and accounted for more than 90% of Cr(VI) mutagenicity. Ternary adducts were also several times more potent in the inhibition of replication than binary complexes. The Cr(VI)-induced mutational spectrum consisted of an approximately equal number of deletions and G/C-targeted point mutations (51% G/C --> T/A and 30% G/C --> A/T). In Escherichia coli cells, Cr(VI)-induced DNA adducts were only highly genotoxic but not mutagenic under either normal or SOS-induced conditions. Lower toxicity and high mutagenicity of ascorbate-Cr(III)-DNA adducts in human cells may result from the recruitment of an error-prone bypass DNA polymerase(s) to the stalled replication forks. Our results suggest that phosphotriester-type DNA adducts could play a more important role in human than bacterial mutagenesis. PMID- 12549929 TI - Structural/functional characterization of the alpha 2-plasmin inhibitor C terminal peptide. AB - The alpha(2)-plasmin inhibitor (A2PI) is a main physiological regulator of the trypsin-like serine proteinase plasmin. It is composed of an N-terminal 15 amino acid fibrin cross-linking polypeptide, a 382-residue serpin domain, and a flexible C-terminal segment. The latter, peptide Asn(398)-Lys(452), and its Lys452Ala mutant were expressed as recombinant proteins in Escherichia coli (r A2PIC and r-A2PICmut, respectively). CD and NMR analyses indicate that r-A2PIC is flexible, loosely folded, and with low content of regular secondary structure. Functional characterization via intrinsic fluorescence ligand titrations shows that r-A2PIC interacts with the isolated plasminogen kringle 1 (r-K1) (K(a) approximately 69.9 mM(-)(1)), K4 (K(a) approximately 45.7 mM(-)(1)), K5 (K(a) approximately 4.3 mM(-)(1)), and r-K2 (K(a) approximately 3.2 mM(-)(1)), all of which are known to exhibit lysine-binding capability. The affinities of these kringles for r-A2PIC are consistently larger than those reported for the ligand N(alpha)-acetyllysine, a mimic of a C-terminal Lys residue. The r-A2PICmut, with a C-terminal Ala residue, also interacts with r-K1 and K4, although with approximately 5-fold lesser affinities relative to r-A2PIC, demonstrating that while Lys(452) plays a major role in the binding, internal residues in r-A2PIC tether the kringles. (1)H NMR spectroscopy shows that key aromatic residues within the K4 lysine-binding site (LBS), namely, Trp(25), Trp(62), Phe(64), Trp(72), and Tyr(74), selectively respond to the addition of r-A2PIC and r A2PICmut, indicating that these interactions proceed via the kringles' canonical LBS. We conclude that r-A2PIC docks to kringles primarily through lysine side chains and that Lys(452) most definitely enhances the binding. This suggests that multiple Lys residues within A2PI could contribute, perhaps in a zipper-like fashion, to its binding to the in-tandem, multikringle array that configures the plasmin heavy chain. PMID- 12549928 TI - Cell surface heparan sulfate participates in CXCL1-induced signaling. AB - The CXC subfamily of chemokines plays an important role in diverse processes, including inflammation, wound healing, growth regulation, angiogenesis, and tumorigenesis. The ELR-CXC chemokine, CXCL1 or MGSA/GROalpha, is traditionally considered to attract neutrophils to sites of inflammation. The non-ELR-CXC chemokine, CXCL10 or IP-10, is chemotactic for monocytes, B cells, and activated T lymphocytes. In addition to its role in leukocyte migration, CXCL10 inhibits the angiogenic functions of the ELR-CXC chemokines as well as bFGF and VEGF. Heparan sulfate proteoglycans (HSPGs) are required for the interaction of bFGF and vEGF ligands and their receptors. However, the role of HSPGs in regulating the ELR-chemokines signaling and biological functions is poorly understood. We show here that the CXCL1 maximal binding to CXCR2 expressed on HEK293 and CHO-K1 cells is dependent on the presence of cell surface HSPGs. The cell surface HSPGs on cells are required for CXCL1-induced PAK1 activation. Moreover, CXCL10 can inhibit CXCL1-induced PAK1 and ERK activation as well as the CXCL1-induced chemotaxis through decreasing CXCL1 binding to cell surface heparan sulfate. These data indicate that HSPGs are involved in modulating CXCL1-induced PAK1 activation and chemotaxis through regulating CXCL1 binding activity to CXCR2 receptor. CXCL10 inhibits CXCL1-induced PAK1 activation and chemotaxis by interfering with appropriate binding of CXCL1 to CXCR2 receptor. PMID- 12549930 TI - Evidence for involvement of the putative first extracellular loop in differential volume sensitivity of the Na+/H+ exchangers NHE1 and NHE2. AB - We studied hyperosmolarity-induced changes in cell volume and cytoplasmic pH in PS120 cells expressing Na(+)/H(+) exchanger (NHE) isoforms and their mutants. Change in cell volume was estimated by measuring change in cell height by means of confocal microscopy. Regulatory volume increase (RVI) and cytoplasmic alkalinization were observed in cells expressing NHE1 but not in cells expressing NHE2 or NHE3. Studies using chimeric exchangers revealed that the membrane domain of the exchanger is responsible for the difference in volume sensitivity between NHE1 and NHE2. Although deletion or point mutation within the first extracellular loop of NHE1 did not affect RVI and alkalinization, point mutations within the corresponding region of NHE2, particularly a region containing aa 41-53, as well as replacement of the N-terminus of NHE2 with the corresponding region of NHE1, rendered NHE2 responsive to the activating effect of cell shrinkage. Thus, the membrane domain plays an important role in the response of the exchanger to cell shrinkage. The data suggest that the putative first extracellular loop of NHE2, but not that of NHE1, may exert an inhibitory influence on hyperosmolarity induced activation of the exchanger and thereby block RVI. PMID- 12549931 TI - Role of glutamate-126 and arginine-144 in the lactose permease of Escherichia coli. AB - Several previous studies have suggested that glutamate-126 and arginine-144 in the lactose permease of Escherichia coli form an ion pair that is essential for sugar binding. To further investigate the role of these residues, E126Q, R144Q, and R144S mutants were made. The R144Q and R144S strains, which had negligible levels of transport, were used as parental strains to isolate suppressor mutations that partially restored sugar transport. The R144Q parent only yielded first-site revertants, but the R144S strain produced three types of second-site replacements: E126Q, V229A, and L330R. In downhill transport assays, the E126Q strain was able to transport lactose at low levels, with an apparent K(m) 3-fold higher than the wild-type strain but a severely depressed apparent V(max). A triple mutant, E126Q/R144S/V229A, showed a relatively robust V(max) value for downhill transport and could actively accumulate lactose against a concentration gradient. Taken together, these results indicate that Glu-126 and Arg-144 are not essential for sugar binding. An alternative explanation for their role in maintaining secondary structure is discussed. PMID- 12549932 TI - Lipopolysaccharides in bacterial membranes act like cholesterol in eukaryotic plasma membranes in providing protection against melittin-induced bilayer lysis. AB - Melittin is a small, cationic peptide that, like many other antimicrobial peptides, lyses cell membranes by acting on their lipid bilayers. However, the sensitivity to antimicrobial peptides varies among cell types. We have performed direct binding and vesicle leakage experiments to determine the sensitivity to melittin of bilayers composed of various physiologically relevant lipids, in particular, key components of eukaryotic membranes (cholesterol) and bacterial outer membranes (lipopolysaccharide or LPS). Melittin binds to bilayers composed of both zwitterionic and negatively charged phospholipids, as well as to the highly charged LPS bilayers. The magnitude of the free energy of binding (deltaG degrees ) increases with increasing bilayer charge density; deltaG degrees = -7.6 kcal/mol for phosphatidylcholine (PC) bilayers and -8.9 to -11.0 kcal/mol for negatively charged bilayers containing phosphatidylserine (PS), phospholipids with covalently attached polyethylene glycol (PEG-lipids), or LPS. Comparisons of these data show that binding is not markedly affected by the steric barrier produced by the PEG in PEG-lipids or by the polysaccharide core of LPS. The addition of equimolar cholesterol to PC bilayers reduces the level of binding (deltaG degrees = -6.4 kcal/mol) and reduces the extent of melittin-induced leakage by 20-fold. LPS and 1:1 PC/cholesterol bilayers have similar high resistance to melittin-induced leakage, indicating that cholesterol in eukaryotic plasma membranes and LPS in Gram-negative bacteria provide strong protection against the lytic effects of melittin. We argue that this resistance is due at least in part to the similar tight packing of the lipid acyl chains in PC/cholesterol and LPS bilayers. The addition of bacterial phospholipids to LPS bilayers increases their sensitivity to melittin, helping to explain the higher sensitivity of deep rough bacteria compared to smooth phenotypes. PMID- 12549933 TI - MnmA and IscS are required for in vitro 2-thiouridine biosynthesis in Escherichia coli. AB - Thionucleosides are uniquely present in tRNA. In many organisms, tRNA specific for Lys, Glu, and Gln contain hypermodified 2-thiouridine (s(2)U) derivatives at wobble position 34. The s(2) group of s(2)U34 stabilizes anticodon structure, confers ribosome binding ability to tRNA and improves reading frame maintenance. Earlier studies have mapped and later identified the mnmA gene (formerly asuE or trmU) as required for the s(2)U modification in Escherichia coli. We have prepared a nonpolar deletion of the mnmA gene and show that it is not required for viability in E. coli. We also cloned mnmA from E. coli, and overproduced and purified the protein. Using a gel mobility shift assay, we show that MnmA binds to unmodified E. coli tRNA(Lys) with affinity in the low micromolar range. MnmA does not bind observably to the nonsubstrate E. coli tRNA(Phe). Corroborating this, tRNA(Glu) protected MnmA from tryptic digestion. ATP also protected MnmA from trypsinolysis, suggesting the presence of an ATP binding site that is consistent with analysis of the amino acid sequence. We have reconstituted the in vitro biosynthesis of s(2)U using unmodified E. coli tRNA(Glu) as a substrate. The activity requires MnmA, Mg-ATP, l-cysteine, and the cysteine desulfurase IscS. HPLC analysis of thiolated tRNA digests using [(35)S]cysteine confirms that the product of the in vitro reaction is s(2)U. As in the case of 4-thiouridine synthesis, purified IscS-persulfide is able to provide sulfur for in vitro s(2)U synthesis in the absence of cysteine. Small RNAs that represent the anticodon stem loops for tRNA(Glu) and tRNA(Lys) are substrates of comparable activity to the full length tRNAs, indicating that the major determinants for substrate recognition are contained within this region. PMID- 12549934 TI - Changes in stability upon charge reversal and neutralization substitution in staphylococcal nuclease are dominated by favorable electrostatic effects. AB - Single site mutations that reverse or neutralize a surface charge were made at 22 ionizable residues in staphylococcal nuclease. Unfolding free energies were obtained by guanidine hydrochloride denaturation. These data, in conjunction with previously obtained stabilities of the corresponding alanine mutants, unequivocally show that the dominant contribution to stability for virtually all of the wild-type side chains examined is the electrostatic effect associated with each residue's charged group. With only a few exceptions, these charges stabilize the native state, with an average loss of 0.5 kcal/mol of stability upon neutralization of a charge. When the charge is reversed, the average destabilization is doubled. Structure-based calculations of electrostatic free energy with the continuum method based on the finite difference solution to the linearized Poisson-Boltzmann equation reproduce the observed energetics when the polarizability in the protein interior is represented with a dielectric constant of 20. However, in some cases, large differences are found, giving insight into possible areas for improvement of the calculations. In particular, it appears that the assumptions made in the calculations about the absence of electrostatic interactions in the denatured state and the energetic consequences of dynamic fluctuations in the native state will have to be further explored. PMID- 12549935 TI - Evaluation of the catalytic mechanism of the p21-activated protein kinase PAK2. AB - The p21-activated kinases (PAKs) play important roles in diverse cellular processes. In the present study, we provide an in-depth kinetic analysis of one of the PAK family members, PAK2, in phosphorylation of a protein substrate, myelin basic protein (MBP), and a synthetic peptide substrate derived from LIM kinase, LIMKtide. Steady-state kinetic analysis of the initial reaction velocity of PAK2 phosphorylation of MBP is consistent with both randomly and compulsorily ordered mechanisms. Further kinetic studies carried out in various concentrations of sucrose revealed that solvent viscosities had no effect on k(cat)/K(m) for either ATP or MBP while k(cat) was highly sensitive to solvent viscosity, indicating that the enzymatic phosphorylation by PAK2 can be best interpreted by a rapid-equilibrium random bi-bi reaction model, and k(cat) is partially limited by both phosphoryl group transfer (31 s(-)(1)) and the product release (19 s( )(1)). In the phosphorylation of LIMKtide, both k(cat) and k(cat)/K(m) were insensitive to solvent viscosity, and the product release (86 s(-)(1)) was much faster than the phosphoryl group transfer step (19 s(-)(1)). These studies suggest that the release of phospho-MBP product is likely partially rate determining for the PAK2-catalyzed reaction since the dissociation rate of products from the PAK2 active site for LIMKtide phosphorylation differs from that of MBP significantly. Such a mechanism is in contrast to the previously established kinetics for the phosphorylation of peptide substrates by cAMP dependent kinase, in which this process is limited by the release of ADP but not the phospho-peptide product. These results complement previous structure-function studies of PAKs and provide important insight for mechanistic interpretation of the kinase functions. PMID- 12549936 TI - Rhodobacter capsulatus 1-deoxy-D-xylulose 5-phosphate synthase: steady-state kinetics and substrate binding. AB - 1-Deoxy-d-xylulose 5-phosphate synthase (DXP synthase) catalyzes the thiamine diphosphate (TPP)-dependent condensation of pyruvate and d-glyceraldehyde 3 phosphate (GAP) to yield DXP in the first step of the methylerythritol phosphate pathway for isoprenoid biosynthesis. Steady-state kinetic constants for DXP synthase calculated from the initial velocities measured at varying concentrations of substrates were as follows: k(cat) = 1.9 +/- 0.1 s(-1), K(m)(GAP) = 0.068 +/- 0.001 mM, and K(m)(pyruvate) = 0.44 +/- 0.05 mM for pyruvate and GAP; k(cat) = 1.7 +/- 0.1 s(-1), K(m)(d-glyceraldehyde) = 33 +/- 3 mM, and K(m)(pyruvate) = 1.9 +/- 0.5 mM for d-glyceraldehyde and pyruvate. beta Fluoropyruvate was investigated as a dead-end inhibitor for pyruvate. Double reciprocal plots showed a competitive inhibition pattern with respect to pyruvate and noncompetitive inhibition with respect to GAP/d-glyceraldehyde. (14)CO(2) trapping experiments demonstrated that the binding of both substrates (pyruvate and GAP/d-glyceraldehyde) is required for the formation of a catalytically competent enzyme-substrate complex. These results are consistent with an ordered mechanism for DXP synthase where pyruvate binds before GAP/d-glyceraldehyde. PMID- 12549937 TI - Characterization of the magnitude and kinetics of xanthine oxidase-catalyzed nitrate reduction: evaluation of its role in nitrite and nitric oxide generation in anoxic tissues. AB - In addition to nitric oxide (NO) generation from specific NO synthases, NO is also formed during anoxia from nitrite reduction, and xanthine oxidase (XO) catalyzes this process. While in tissues and blood high nitrate levels are present, questions remain regarding whether nitrate is also a source of NO and if XO-mediated nitrate reduction can be an important source of NO in biological systems. To characterize the kinetics, magnitude, and mechanism of XO-mediated nitrate reduction under anaerobic conditions, EPR, chemiluminescence NO-analyzer, and NO-electrode studies were performed. Typical XO reducing substrates, xanthine, NADH, and 2,3-dihydroxybenz-aldehyde, triggered nitrate reduction to nitrite and NO. The rate of nitrite production followed Michaelis-Menten kinetics, while NO generation rates increased linearly following the accumulation of nitrite, suggesting stepwise-reduction of nitrate to nitrite then to NO. The molybdenum-binding XO inhibitor, oxypurinol, inhibited both nitrite and NO production, indicating that nitrate reduction occurs at the molybdenum site. At higher xanthine concentrations, partial inhibition was seen, suggesting formation of a substrate-bound reduced enzyme complex with xanthine blocking the molybdenum site. The pH dependence of nitrite and NO formation indicate that XO-mediated nitrate reduction occurs via an acid-catalyzed mechanism. With conditions occurring during ischemia, myocardial xanthine oxidoreductase and nitrate levels were determined to generate up to 20 microM nitrite within 10-20 min that can be further reduced to NO with rates comparable to those of maximally activated NOS. Thus, XOR catalyzed nitrate reduction to nitrite and NO occurs and can be an important source of NO production in ischemic tissues. PMID- 12549938 TI - The initiating steps of a type II fatty acid synthase in Plasmodium falciparum are catalyzed by pfACP, pfMCAT, and pfKASIII. AB - Malaria, a disease caused by protozoan parasites of the genus Plasmodium, is one of the most dangerous infectious diseases, claiming millions of lives and infecting hundreds of millions of people annually. The pressing need for new antimalarials has been answered by the discovery of new drug targets from the malaria genome project. One of the early findings was the discovery of two genes encoding Type II fatty acid biosynthesis proteins: ACP (acyl carrier protein) and KASIII (beta-ketoacyl-ACP synthase III). The initiating steps of a Type II system require a third protein: malonyl-coenzyme A:ACP transacylase (MCAT). Here we report the identification of a single gene from P. falciparum encoding pfMCAT and the functional characterization of this enzyme. Pure recombinant pfMCAT catalyzes malonyl transfer from malonyl-coenzyme A (malonyl-CoA) to pfACP. In contrast, pfACP(trans), a construct of pfACP containing an amino-terminal apicoplast transit peptide, was not a substrate for pfMCAT. The product of the pfMCAT reaction, malonyl-pfACP, is a substrate for pfKASIII, which catalyzes the decarboxylative condensation of malonyl-pfACP and various acyl-CoAs. Consistent with a role in de novo fatty acid biosynthesis, pfKASIII exhibited typical KAS (beta-ketoacyl ACP synthase) activity using acetyl-CoA as substrate (k(cat) 230 min(-1), K(M) 17.9 +/- 3.4 microM). The pfKASIII can also catalyze the condensation of malonyl-pfACP and butyryl-CoA (k(cat) 200 min(-1), K(M) 35.7 +/- 4.4 microM) with similar efficiency, whereas isobutyryl-CoA is a poor substrate and displayed 13-fold less activity than that observed for acetyl-CoA. The pfKASIII has little preference for malonyl-pfACP (k(cat)/K(M) 64.9 min(-1)microM( 1)) over E. coli malonyl-ACP (k(cat)/K(M) 44.8 min(-1)microM(-1)). The pfKASIII also catalyzes the acyl-CoA:ACP transacylase (ACAT) reaction typically exhibited by KASIII enzymes, but does so almost 700-fold slower than the KAS reaction. Thiolactomycin did not inhbit pfKASIII (IC(50) > 330 microM), but three structurally similar substituted 1,2-dithiole-3-one compounds did inhibit pfKASIII with IC(50) values between 0.53 microM and 10.4 microM. These compounds also inhibited the growth of P. falciparum in culture. PMID- 12549939 TI - Stoichiometry and affinity of nucleotide binding to P-glycoprotein during the catalytic cycle. AB - Drug transport mediated by P-glycoprotein (Pgp) is driven by hydrolysis of ATP at the two cytosolic nucleotide binding domains. However, little is currently known concerning the stoichiometry of nucleotide binding and how both stoichiometry and binding affinity change during the catalytic cycle of the transporter. To address this issue, we used fluorescence techniques to measure both the number of nucleotides bound to P-glycoprotein during various stages of the catalytic cycle and the affinity of nucleotide binding. Results showed that resting state P glycoprotein bound two molecules of the fluorescent nucleotide derivative, 2'(3') O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate (TNP-ATP), whereas the vanadate trapped transition state bound only one nucleotide molecule. Both resting and transition state P-glycoprotein showed similar affinity for TNP-ATP/TNP-ADP and unlabeled ATP/ADP. Following binding of various drugs, resting state P glycoprotein displayed a higher affinity for nucleotides, up to 4-fold depending on the compound used. In contrast, the transition state showed substantially lower (up to 3-fold) nucleotide binding affinity when the drug binding site(s) is/are occupied. These results indicate that both nucleotide binding domains of P glycoprotein are likely to be occupied with either ATP (or ADP) in the resting state and the transition state in the absence of transport substrates. Drugs alter the binding affinity to favor association of ATP with P-glycoprotein at the start of the catalytic cycle and release of ADP from the transition state following nucleotide hydrolysis. PMID- 12549941 TI - Failure of cetirizine and fexofenadine to prevent motion sickness. AB - OBJECTIVE: To determine the effectiveness of 2 second-generation antihistamines in modulating motion sickness induced by Coriolis vestibular cross-coupling stimulation. METHODS: This prospective, randomized, double-blind, crossover, placebo-controlled study was conducted in 18 healthy adults. Subjects were exposed to Coriolis vestibular cross-coupling in the laboratory using the Staircase Profile Test for baseline susceptibility and when under the influence of cetizirine, fexofenadine, and placebo. Subjective evaluation of sickness symptoms was based on the Graybiel diagnostic criteria of acute motion sickness, Golding's scale, and the Coriolis Sickness Susceptibility Index. RESULTS: Repeated measures ANOVA and Friedman nonparametric ANOVA of rank tests revealed that there were significant differences in symptom assessments based on Graybiel's diagnostic criteria (p < or = 0.001), subjective symptoms of motion sickness (p < or = 0.001), and state-anxiety (p < or = 0.001) before and after motion exposure. However, there are no significant differences between the baseline susceptibility to motion sickness and treatment with placebo, cetirizine, or fexofenadine. CONCLUSIONS: The failure of the second-generation antihistamines cetirizine and fexofenadine to prevent motion sickness suggests that the therapeutic actions of this class of antihistamines against motion sickness may be mediated through central versus peripheral receptors. The sedative effect of other antihistamines, such as hydroxyzine, may play a more significant role in alleviating motion sickness than previously thought. PMID- 12549942 TI - Effect of clarithromycin on steady-state digoxin concentrations. AB - OBJECTIVE: To evaluate the magnitude and dose-relatedness of the effect of clarithromycin on the pharmacokinetics of digoxin, and to compare the effects of clarithromycin with those of P-glycoprotein inhibitors. METHODS: Eight Japanese inpatients with congestive heart failure participated in this study. Each patient received oral digoxin therapy for at least 7 days and were coadministered oral clarithromycin to prevent or treat pneumonia. To evaluate the effects of clarithromycin on the pharmacokinetics of digoxin, digoxin concentrations were compared before and after coadministration of clarithromycin. RESULTS: Digoxin concentrations were higher after coadministration of clarithromycin in all patients (before, 0.838 +/- 0.329 ng/mL; after, 1.36 +/- 0.619 ng/mL); (p < 0.005). A significant correlation was observed between the dose of clarithromycin and the percentage of increase in the digoxin concentration. CONCLUSIONS: Digoxin concentrations increased during concomitant administration of clarithromycin, and this effect was dose-dependent on clarithromycin. The percentage increase in digoxin concentrations after the usual oral dose of clarithromycin (400 mg/d) is approximately 70%. Therefore, digoxin concentrations must be monitored carefully after coadministration of clarithromycin, and the doses of digoxin may need readjustment in patients who are concomitantly receiving clarithromycin. PMID- 12549943 TI - Aminoglycoside-associated nephrotoxicity in the elderly. AB - OBJECTIVE: To determine whether there was an increased incidence of nephrotoxicity in elderly patients (> or =65 y) prescribed single-dose (SD) versus multiple-dose (MD) aminoglycosides and whether aminoglycoside-induced nephrotoxicity was associated with length of therapy and other risk factors. METHODS: A prospective, observational audit at a university teaching hospital was conducted. Physician prescribing was used to stratify subjects according to dosing regimen: MD (n = 60) or SD (n = 26). Nephrotoxicity was defined as an increase in the serum creatinine level of 0.5 mg/dL sustained over 2 days. RESULTS: Eighty-six patients were included; 9.3% developed nephrotoxicity, of whom 62.5% received SD therapy. The incidence of nephrotoxicity did not differ between regimens (p = 0.051). There was an increased length of therapy in those who developed nephrotoxicity (mean +/- SD 6.1 +/- 6.2 vs. 3.7 +/- 2.8 d; p = 0.044). Additionally, patients who developed nephrotoxicity had an increased length of hospitalization (20.3 +/- 16.1 vs. 8.4 +/- 5.4 d; p < 0.001). Nephrotoxicity correlated with a diagnosis of diabetes mellitus (OR 15.1; 95% CI 1.11 to 205), concomitant angiotensin-converting enzyme (ACE) inhibitor therapy (OR 28.0; 95% CI 2.15 to 365), and SD therapy (OR 20.7; 95% CI 1.45 to 297). CONCLUSIONS: Our overall incidence of nephrotoxicity is consistent with that reported in the literature. A diagnosis of diabetes mellitus, concomitant use of ACE inhibitors, and SD regimens were risk factors for the development of nephrotoxicity. An adequately powered, randomized trial is needed to assess whether a difference in the incidence of nephrotoxicity exists between SD and MD therapy in the elderly. PMID- 12549944 TI - Pharmacy-based intervention to reduce antibiotic use for acute bronchitis. AB - BACKGROUND: Intervention programs can reduce inappropriate antibiotic use for the treatment of acute bronchitis in a closed health maintenance organization model. OBJECTIVE: To evaluate the impact of a pharmacy-based intervention program intended to reduce antibiotic use in the treatment of acute bronchitis in a community-based physician group model. SUBJECTS: Adult and pediatric patients with an office or urgent care visit for acute bronchitis during the baseline and study periods were included in the study. The clinicians were primary care physicians, nurse practitioners, and physician assistants in a suburban community based physician group setting. METHODS: All patients treated for acute bronchitis from January 1 through June 30, 1998, were evaluated for initial receipt of antibiotics and use of clinic resources (office visits, additional antibiotics). From September through December of 1998, physicians were provided literature from the Centers for Disease Control and Prevention (CDC), cough and cold package inserts, and newsletters intended to educate the providers regarding the inappropriateness of antibiotics in the treatment of acute bronchitis. Patient directed literature from the CDC was placed in the examination rooms and clinic waiting areas beginning September 1998. From January 1 through June 30, 1999, all patients treated for acute bronchitis were assessed for receipt of antibiotics and use of clinic resources. A separate geographic clinic site served as a control during both study periods. RESULTS: During 1998, 888 of 1840 patients (48.3%) received antibiotics for treatment of acute bronchitis; this total decreased to 924 of 2392 (38.6%; p < or = 0.001) in 1999, a reduction of 20%. The rate of antibiotic prescribing in control patients was unchanged during the concomitant time periods (142/446, 31.8% vs. 102/321, 31.8%). The rate of subsequent physician visits was similar (8% vs. 9%) between patients receiving antibiotics and those who did not. However, significantly more patients initially receiving antibiotics required a subsequent antibiotic prescription (45/1812, 2.5% vs. 24/2420, 1.0%; p < or = 0.001). CONCLUSIONS: A pharmacy-based intervention program reduces the incidence of inappropriate antibiotic use in the treatment of acute bronchitis. Reduced antibiotic prescribing does not increase consumption of healthcare resources; patients who receive antibiotics for acute bronchitis are more likely to subsequently require additional antibiotic prescriptions. While a significant decrease in antibiotic use was realized, other interventions are required to further reduce the prevalence of antibiotic use in acute bronchitis. PMID- 12549945 TI - Antifungal susceptibility testing in teaching hospitals. AB - BACKGROUND: An assessment of antifungal susceptibility testing (AST) has not been conducted since the introduction of the National Committee for Clinical Laboratory Standards (NCCLS) M27-A document. OBJECTIVE: To determine AST practices in teaching hospitals. METHODS: A questionnaire was mailed to the heads of 386 randomly assigned microbiology departments from teaching hospitals identified through the 2000 American Hospital Association Guide. Identifiers were used to delineate responders from nonresponders. A reminder letter was mailed 3 weeks after the initial mailing to all nonresponders. The hospital bed-size and number of inpatient days for respondents were obtained through the American Hospital Directory. RESULTS: The questionnaire was returned by 171 (44.3%) institutions. The total and median (range) number of candida isolates were 137,088 and 8.5 (1-145)/1000 inpatient days for the year 2000, respectively. Approximately 1% (1300) of candida isolates, from predominantly blood specimens, underwent AST. AST was reported by 115 (67.2%) hospitals, with testing on site at 27 hospitals and off site for 88 hospitals. NCCLS methodology (80% broth microdilution) was used by 75% of the hospitals performing on-site AST. The median time to obtain AST results was significantly lower when testing was performed on site (3 d) compared with off site (7-10 d). SUMMARY: A large number of candida bloodstream isolates undergoes AST annually. AST results are obtained sooner when performed on site compared with off site. PMID- 12549946 TI - Faculty turnover within academic pharmacy departments. AB - BACKGROUND: Pharmacy faculty manpower has been debated within the academic pharmacy community over the last several decades. Previous investigations studied job satisfaction among faculty members, but have not evaluated faculty retention and turnover among academic pharmacy departments. OBJECTIVE: To evaluate retention and turnover rates in the departments of Pharmacy Practice and Basic Science (Pharmacology/Toxicology, Pharmaceutics, Medicinal Chemistry) over the last 5 years. METHODS: Individual instructors and assistant, associate, and full professors across 80 colleges of pharmacy in the US were tracked between the years 1996 and 2001 using the American Association of Colleges of Pharmacy published rosters. Differences between departments were analyzed by year stratified cross-tabulation table analysis. RESULTS: A greater percentage of Pharmacy Practice faculty resigned (10.6%) compared with Basic Science faculty (6.0%; percent ratio 1.76; 95% CI 1.58 to 1.95; p < 0.001), which remained constant across each academic year. Approximately 2.7 faculty members left their academic institutions per year in Pharmacy Practice compared with 1.1 faculty members in the aggregate of Basic Science departments. A higher percentage of women resigned in Pharmacy Practice (13.2%) than did men (8.7%; percent ratio 1.5; 95% CI 1.34 to 1.68; p < 0.001), despite a 1.3-fold male to female ratio. Likewise, regardless of a 4.1-fold male to female ratio in the Basic Science group, a higher percentage of women resigned (8.0%) than men (5.5%; percent ratio 1.45; 95% CI 1.18 to 1.78; p < 0.001). CONCLUSIONS: Over a 5-year period, Pharmacy Practice exhibited a higher turnover compared with Basic Science. Women displayed significantly higher turnover than men across all pharmacy academic departments. New retention approaches, especially for female faculty members, should be explored. PMID- 12549947 TI - Pseudo-Parkinson disease secondary to ritonavir-buspirone interaction. AB - OBJECTIVE: To report a case of Parkinson-like symptoms appearing in a patient after introduction of ritonavir to buspirone therapy. CASE SUMMARY: A 54-year-old HIV-positive white man presented to the clinic with a 2-week history of ataxia, shuffling gait, cogwheel rigidity, resting tremor, and sad affect with masked features. This patient had been receiving high-dose buspirone (40 mg every morning and 30 mg every evening) for 2 years prior to the introduction of ritonavir/indinavir combination therapy (400 mg/400 mg twice daily) 6 weeks prior to initiation of the above symptoms. Buspirone was decreased to 15 mg 3 times daily, ritonavir/indinavir was discontinued, and amprenavir 1200 mg twice daily was added. The patient's symptoms began to subside after 1 week, with complete resolution after about 2 weeks. The patient continued to receive buspirone for an additional 12 months without recurrence of symptoms. DISCUSSION: This is the first reported interaction of buspirone and antiretrovirals. Buspirone, extensively metabolized by CYP3A4, was likely at supratherapeutic levels due to the inhibitory effect of ritonavir and, secondarily, indinavir. The Parkinson like symptoms developed rapidly and severely, impacted this patient's quality of life, and necessitated significant clinic expenditures to identify this drug-drug interaction. CONCLUSIONS: This case demonstrates a severe drug-drug interaction between buspirone and ritonavir and further demonstrates the need for awareness of the metabolic profile for all agents an HIV-infected patient is receiving. PMID- 12549948 TI - Resolution of risperidone-induced hyperprolactinemia with substitution of quetiapine. AB - OBJECTIVE: To report a case of risperidone-induced hyperprolactinemia that was successfully managed with quetiapine. CASE SUMMARY: A 30-year-old white woman with schizoaffective disorder, depressive type, and comorbid alcohol and cocaine abuse was treated successfully for her psychotic symptoms with risperidone until she developed adverse effects consistent with hyperprolactinemia. This was confirmed by laboratory blood tests, as her prolactin level was 186.9 ng/mL (normal for nonpregnant women 2.8-29.2). The woman had experienced similar effects in the past, which had led to noncompliance and subsequent psychotic relapse. Normalization of prolactin levels and associated adverse effects were achieved upon switching to quetiapine. No psychotic symptoms reoccurred. DISCUSSION: Dopamine type 2 (D(2)) receptor blockade in the mesolimbic tract is thought to mediate the therapeutic effects of antipsychotics. This action in the tuberoinfundibular system produces prolactin level elevation. Risperidone has a relatively higher affinity for the D(2) receptor in comparison with other atypical antipsychotics, which may explain why it is associated with a higher incidence of hyperprolactinemia. Quetiapine, which has one of the lowest D(2) receptor affinities, is not known to increase prolactin levels to any significant degree. This pharmacologic property allows quetiapine to be a reasonable treatment option for patients who develop risperidone-induced hyperprolactinemia. CONCLUSIONS: Quetiapine may be a suitable substitute when a patient taking risperidone develops hyperprolactinemia. PMID- 12549949 TI - Serotonin syndrome induced by low-dose venlafaxine. AB - OBJECTIVE: To report the case of a patient with serotonin syndrome induced by low dose venlafaxine. CASE SUMMARY: A 29-year-old Taiwanese woman with major depressive disorder abruptly developed serotonin syndrome during low-dose (37.5 mg/d) venlafaxine monotherapy, with symptoms of restlessness, tremor, shivering, diarrhea, vomiting, ataxia, tachycardia, and myoclonus. The patient recovered in 2 hours after receiving prochlorperazine and lorazepam in the emergency department. Venlafaxine was discontinued, and she was discharged home. Two weeks later, the patient started to receive fluoxetine 20 mg/d and reported no adverse adverse effects during follow-up clinic visits. DISCUSSION: The clinical manifestations of this case meet Sternbach's criteria of serotonin syndrome. Its possible etiologic factors include panic attack, adverse drug reaction, pharmacodynamic interaction, and congenital absence of CYP2D6 enzyme activity. The Naranjo probability scale suggested a probable causality of venlafaxine treatment and serotonin syndrome. CONCLUSIONS: Clinicians should be aware of the risk of serotonin syndrome when the patient receives not only a combination of 2 antidepressants, but also the single potent serotonergic agent venlafaxine. PMID- 12549950 TI - Fenofibrate potentiates warfarin effects. AB - OBJECTIVE: To describe 2 patients in whom the initiation of fenofibrate potentiated warfarin's anticoagulant effects. CASE SUMMARY: A 71-year-old white woman and an 80-year-old white woman with multiple medical conditions were both stabilized on long-term warfarin therapy. During the course of anticoagulation, both patients were prescribed fenofibrate and experienced threefold and twofold increases in international normalized ratio (INR), respectively, requiring total weekly warfarin dosage reductions of 30-40%. Before starting fenofibrate therapy, both patients' coagulation values were within the therapeutic range. When interviewed, patients and caregivers denied bleeding, bruising, changes in diet, alcohol ingestion, nonadherence with therapy, or changes in drug regimen except for the addition of fenofibrate. Upon chart review, evaluation of potentially contributory parameters, such as other changes in drug therapy, thyroid function, liver function, and drug-disease interactions, showed that these parameters remained stable and were ruled noncontributory. DISCUSSION: The addition of fenofibrate in 2 patients on stable and therapeutic doses of warfarin increased the anticoagulant response to warfarin. A clear temporal relationship with the addition of fenofibrate and the appearance of the interaction was seen. Fenofibrate is highly protein bound, with the potential to displace warfarin from its binding protein, leading to an enhanced hypoprothrombinemic effect. Fenofibrate is also a mild to moderate inhibitor of CYP2C9, the enzyme responsible for warfarin metabolism. The combination of these effects- displacement of warfarin by fenofibrate coupled with decreased metabolism of warfarin--may increase the anticoagulant response to warfarin. Using the Naranjo probability scale, these interactions were designated as probable. CONCLUSIONS: We suggest serial monitoring of INR and consider an empiric 20% reduction in warfarin dosage when fenofibrate is initiated, with the possibility for a greater warfarin dosage reduction based on INR results. PMID- 12549951 TI - Clopidogrel-associated leukopenia. AB - OBJECTIVE: To report a case of leukopenia in a patient receiving clopidogrel following intracoronary stent placement. CASE SUMMARY: A 58-year-old white man presented to the emergency department (ED) with fever and chills. Seven days prior to presentation, he underwent intracoronary stent placement and clopidogrel 75 mg once daily was initiated. In the ED, laboratory evaluation revealed leukopenia with negative blood and urine cultures. The patient was released but returned the following day with fever and sweating. Infectious complications were ruled out, and clopidogrel was discontinued. Leukopenia resolved within 7 days, and the patient has experienced no further hematologic complications. Based on the Naranjo probability scale, the leukopenia was rated as a probable reaction to clopidogrel. DISCUSSION: Ticlopidine and clopidogrel are the 2 commercially available thienopyridine antiplatelet agents. Risk of myelotoxicity with ticlopidine is well known and requires frequent hematologic monitoring. Clopidogrel is better tolerated, less toxic, and has more extensive indications, including use in acute coronary syndrome. These factors have resulted in the replacement of ticlopidine by clopidogrel. The 4 previously published reports of clopidogrel-associated white blood cell (WBC) toxicity are described. The mechanism of myelotoxicity with clopidogrel is not known. CONCLUSIONS: Better tolerability, reduced toxicity (particularly hematologic), and the convenience of once-daily dosing have resulted in the replacement of ticlopidine with clopidogrel. WBC toxicity associated with clopidogrel therapy is very infrequent and has occurred weeks to months following therapy initiation. Clinicians should be aware of the potential for this adverse effect. PMID- 12549952 TI - Acute liver failure with concurrent bupropion and carbimazole therapy. AB - OBJECTIVE: To report a case of fatal liver failure possibly associated with concurrent use of bupropion and carbimazole. CASE SUMMARY: A 41-year-old Chinese man with a history of hyperthyroidism had been treated with carbimazole and propranolol for the past 5 years. He received a 10-day course of bupropion as an aid for smoking cessation 10 weeks prior to presentation. He developed acute liver failure with rapid deterioration of renal function. Liver biopsy showed evidence of nonspecific drug-induced acute liver injury. His condition was further complicated by sepsis and coagulopathy. Death resulted 19 days after the onset of symptoms. The likelihood that bupropion induced hepatotoxicity in our patient was possible, based on the Naranjo probability scale. DISCUSSION: Although there is increasing evidence of hepatotoxicity induced by bupropion, this is the first case of fatality that could have resulted from acute liver failure in a patient receiving bupropion while on concomitant treatment with carbimazole. CONCLUSIONS: Clinicians should be aware of the possibility of acute liver insult induced by bupropion given concurrently with other hepatotoxic drugs. PMID- 12549953 TI - Fulminant hepatic failure associated with propylthiouracil. AB - OBJECTIVE: To report 2 fatal cases of fulminant hepatic failure associated with propylthiouracil treatment against hyperthyroidism. CASE SUMMARY: Two women, 30 and 32 years old with no previous liver disease, were treated with propylthiouracil against Graves' disease. Both patients developed jaundice after a 4- and 5-month treatment period, respectively. The disease was similar to viral hepatitis, with a progressive course to severe liver dysfunction and death, along with multisystem organ failure despite extensive therapeutic measures. One of the patients was pregnant and subsequently miscarried. Neither patient had a history of alcoholism, drug abuse, blood transfusion, or exposure to hepatitis A, B, or C. Extrahepatic obstruction was ruled out with an abdominal ultrasonogram. Serologic studies and immunologic tests were negative. A submassive necrosis was shown in a postmortem histologic study. DISCUSSION: Naranjo probability scale criteria applied to both cases confirm the adverse reactions as probable. These cases fit the requirements of drug hepatotoxicity proposed by Hanson and the Council of the International Organization of Medical Sciences. Eight deaths associated to propylthiouracil were found in our review of the medical literature up to December 2000. CONCLUSIONS: Despite the widespread use of propylthiouracil, fulminant hepatitis with death is exceptionally rare; these 2 cases could be added to the fatal outcomes published to date. PMID- 12549954 TI - Lepirudin use in a neonate with heparin-induced thrombocytopenia. AB - OBJECTIVE: To describe a case of heparin-induced thrombocytopenia (HIT) in a premature infant and the doses of danaparoid and lepirudin needed to achieve appropriate therapeutic endpoints. CASE SUMMARY: A 30-week gestational age infant was diagnosed with HIT with heparin antibodies. Danaparoid 2.0-2.4 units/kg/h achieved anti-Xa levels of 0.2-0.4 U/mL, but thrombocytopenia failed to resolve. Lepirudin was started in place of danaparoid. Lepirudin doses of 0.03-0.05 mg/kg/h achieved target activated partial thromboplastin time values of 1.5-2.0 times baseline. DISCUSSION: Dosing information for danaparoid in neonates is limited, and information for lepirudin appears only in German literature at this time. HIT is well documented in newborns, and lepirudin use in these situations is likely to increase. This report provides some guidance for optimal dosing. It also provides some guidance for HIT evaluation in preterm infants, in whom blood volume for laboratory tests is a major issue. CONCLUSIONS: HIT is an important and potentially fatal problem in neonates. Lepirudin may be the drug of choice, especially since danaparoid is now unavailable. Initial lepirudin dosing should not exceed 0.05 mg/kg/h. PMID- 12549955 TI - Macular exanthema due to fumaric acid esters. AB - OBJECTIVE: To report a case of an unusual skin reaction to fumaric acid esters (FAE). CASE SUMMARY: A 68-year-old white woman who was treated with FAE for 4 days for lichen planus developed generalized pruritic exanthema. This was suspected to be an allergic drug reaction to FAE, and the treatment was discontinued. After 48-72 hours, the exanthema resolved completely. An objective causality assessment revealed that the adverse drug event was probable. As skin testing for diagnostic purposes is not feasible with FAE, the drug-related origin of the exanthema was confirmed by oral rechallenge with FAE. DISCUSSION: The effectiveness of FAE in the systemic treatment of psoriasis vulgaris has been proven by controlled clinical trials. The compound has been shown to be tolerable and safe even during prolonged treatment. The most frequent adverse effects are gastrointestinal symptoms and flushing, which typically occur 4-6 hours after administration of the drug. Allergic reactions to FAE have not yet been reported. Since the patient was rechallenged with the suspected drug, we could confirm the allergic origin of the exanthema. CONCLUSIONS: The occurrence of allergic skin reaction should be considered in patients receiving treatment with FAE. PMID- 12549956 TI - Desloratadine: a nonsedating antihistamine. AB - OBJECTIVE: To review information on desloratadine, a nonsedating antihistamine. DATA SOURCES: An English-language MEDLINE search was conducted (1966-July 2002). References of identified articles were subsequently reviewed for additional data. Schering Corporation provided unpublished information. STUDY SELECTION/DATA EXTRACTION: Articles and abstracts pertaining to desloratadine were considered for inclusion, with emphasis on randomized, placebo-controlled, double-blind trials. DATA SYNTHESIS: Desloratadine is approved for the treatment of symptoms associated with seasonal allergic rhinitis (SAR), perennial allergic rhinitis (PAR), and chronic idiopathic urticaria (CIU) in patients aged > or =12 years. In placebo-controlled trials, desloratadine demonstrated superior efficacy as a once daily treatment of SAR, PAR, and CIU. Data suggest that desloratadine has antiinflammatory and decongestant activity. CONCLUSIONS: Desloratadine appears to be a "me-too" agent, with no major differences compared with other second generation antihistamines. PMID- 12549957 TI - Nesiritide: a novel approach for acute heart failure. AB - OBJECTIVE: To review preclinical and clinical information related to nesiritide, a recombinant form of B-type natriuretic peptide approved for treatment of acutely decompensated heart failure. DATA SOURCES: Primary and review articles were identified by MEDLINE search (1966-May 2002) using the key words natriuretic peptide and heart failure, and through secondary sources. Natrecor's document submitted for the Food and Drug Administration (FDA) New Drug Application were obtained from the FDA Web site. STUDY SELECTION/DATA EXTRACTION: Peer-reviewed articles and abstracts of randomized clinical trials in humans were included in this review. DATA SYNTHESIS: Nesiritide has beneficial actions for treatment of heart failure, including arterial and venous dilatation, enhanced sodium and urinary excretion, and suppression of the renin-angiotensin-aldosterone and sympathetic nervous systems. It has been shown to improve hemodynamic parameters, primarily pulmonary capillary wedge pressure, as well as clinical symptoms in patients with acutely decompensated heart failure. Nesiritide produced more rapid hemodynamic improvement and caused significantly fewer adverse effects than intravenous nitroglycerin. The incidence of hypotension, the most common adverse effect, was comparable between nesiritide and nitroglycerin. Additionally, nesiritide is associated with a lower incidence of arrhythmias than dobutamine and has a neutral effect on mortality. CONCLUSIONS: Nesiritide offers an alternative for management of acutely decompensated heart failure. It is considered an option for patients who do not respond to other vasodilators, inotropes, or diuretics and for those at high risk of arrhythmias. Further pharmacoeconomic investigations for nesiritide are warranted. PMID- 12549958 TI - Factors influencing consumer use of written drug information. AB - OBJECTIVE: To provide an overview of the use and impact of written drug information (WDI) on consumers, and to review the literature on the factors influencing the use of WDI by consumers. DATA SOURCES: Relevant articles published in English since the late 1970s were identified based on searches of on line databases, texts, and cited references in published articles. STUDY SELECTION: Articles reporting findings on the origin, use, and impact of WDI were included. Due to limited literature, articles reporting findings on factors influencing the use of written drug as well as disease information were included. DATA EXTRACTION: Due to the lack of design consistency between studies and the comparatively small volume of work, subjective assessment rather than a criteria based objective review was deemed more appropriate. DATA SYNTHESIS: To date, research on WDI has focused on its use and impact. WDI has the potential to increase patients' knowledge, compliance, and satisfaction. However, there is also the potential for anxiety or premature cessation of therapy due to fear of possible adverse effects. Multiple factors may potentially influence the use of WDI by consumers including those associated with the written information document (readability, presentation), the patient (health literacy, role of caregiver, demographic factors, health locus of control, coping style, health belief model), and the environment (timing of provision, experience). CONCLUSIONS: WDI has the potential to impact consumers positively and negatively. Although not widely investigated, a number of factors can potentially influence the use of WDI by consumers. The findings of this review can form the basis for much needed further research. PMID- 12549959 TI - Tamoxifen for prevention of breast cancer. AB - OBJECTIVE: To review data on the use of tamoxifen for primary prevention of breast cancer. DATA SOURCES: A literature search was performed through MEDLINE (1992-May 2002) using the key words tamoxifen, breast cancer, and prevention. DATA SYNTHESIS: Breast cancer is the most common malignancy detected in American women. Attempts to reduce morbidity and mortality include early detection programs and chemoprevention. Clinical trials of tamoxifen for reduction of breast cancer risk are reviewed. CONCLUSIONS: Tamoxifen may reduce the risk of primary breast cancer in women at increased risk. The benefit of tamoxifen in women who are not at risk is uncertain. The risks of developing thromboembolic disorders or endometrial cancer must be considered before tamoxifen is prescribed. Women should be given all of the information about the benefits and risks of tamoxifen use so that they can make an informed decision based on the best data available. PMID- 12549960 TI - Association of HMG-CoA reductase inhibitors with neuropathy. AB - OBJECTIVE: To review the possible association between statins and peripheral neuropathy. DATA SOURCES: Literature was obtained from MEDLINE (1984-September 2002) and International Pharmaceutical Abstracts (1970-June 2002). Key search terms included statin, neuropathy, and HMG-CoA reductase inhibitor. DATA SYNTHESIS: Epidemiologic studies and case reports suggest an increased risk of peripheral neuropathy with statin drugs. Most patients were receiving long-term therapy, although the onset was highly variable. The majority of cases were at least partially reversible with drug cessation. Specific risk factors or mechanisms have not been identified. CONCLUSIONS: Observational data suggest a link between chronic statin use and increased risk of peripheral neuropathy. However, the risk appears to be small relative to the significant cardioprotective benefits. PMID- 12549961 TI - Ivermectin for the treatment of resistant scabies. AB - OBJECTIVE: To assess the clinical literature describing the use of ivermectin in cases of treatment-resistant scabies infestations. DATA SOURCES: Clinical literature accessed through a MEDLINE search (1966-July 2001) and the Cochrane Database. DATA SYNTHESIS: A limited number of case reports and case series describes successful intervention with ivermectin use in patients who have failed prior therapy. Ivermectin may serve as a viable alternative therapy as the incidence of resistant scabitic infestations increases. CONCLUSIONS: Ivermectin appears to be a safe and effective alternative for patients with treatment resistant scabies. Larger, controlled clinical trials are required before this therapy can be recommended in the general population or as primary therapy. PMID- 12549962 TI - Gabapentin for the treatment of tremor. AB - OBJECTIVE: To review the use of the antiepileptic drug gabapentin for the treatment of various types of tremor. DATA SOURCES: A search of biomedical literature was completed through MEDLINE and EMBASE (1993-May 2002) to identify all clinical trials pertaining to the use of gabapentin for the treatment of tremor in humans. DATA SYNTHESIS: Outcome data from the few published studies have varied widely. Patient groups have been small, and conclusions have been based largely on subjective patient and investigator ratings. CONCLUSIONS: A trial of gabapentin is warranted in patients who fail therapy with traditional agents. Improvement should be measured by a patient's perceived functional ability. PMID- 12549963 TI - Personal digital assistant applications for the healthcare provider. AB - OBJECTIVE: To review some common medical applications available for personal digital assistants (PDAs), with brief discussion of the different PDA operating systems and memory requirements. Key search terms included handheld, PDA, personal digital assistants, and medical applications. DATA SOURCES: The literature was accessed through MEDLINE (1999-August 2002). Other information was obtained through secondary sources such as Web sites describing common PDAs. DATA SYNTHESIS: Medical applications available on PDAs are numerous and include general drug references, specialized drug references (e.g., pediatrics, geriatrics, cardiology, infectious disease), diagnostic guides, medical calculators, herbal medication references, nursing references, toxicology references, and patient tracking databases. Costs and memory requirements for these programs can vary; consequently, the healthcare provider must limit the medication applications that are placed on the handheld computer. CONCLUSIONS: This article attempts to systematically describe the common medical applications available for the handheld computer along with cost, memory and download requirements, and Web site information. This review found many excellent PDA drug information applications offering many features which will aid the healthcare provider. Very likely, after using these PDA applications, the healthcare provider will find them indispensable, as their multifunctional capabilities can save time, improve accuracy, and allow for general business procedures as well as being a quick reference tool. To avoid the benefits of this technology might be a step backward. PMID- 12549964 TI - Figs--the earliest known ancient drug for cutaneous anthrax. AB - BACKGROUND: Anthrax is an often fatal bacterial infection, occurring in cutaneous, inhalational, gastrointestinal, and meningeal forms. Evaluation of anthrax treatment from ancient history may help healthcare providers to handle this serious disease more efficiently. OBJECTIVE: To evaluate the biblical descriptions of anthrax, focusing on its therapy in ancient times. STUDY SELECTION: All biblical texts associated with anthrax were examined and passages relating to this disease were studied closely. DATA SYNTHESIS: Biblical passages such as: "Take a cluster of figs. And they took and laid it on the boil [anthrax], and he recovered," and "Let them take a cluster of figs, and lay it upon the boil [anthrax], and he shall recover," convincingly indicate that figs have healing properties in cutaneous anthrax lesions. CONCLUSIONS: On the basis of this study, the National Institutes of Health together with the pharmaceutical industry may consider to work on isolation of compounds from fig plants to develop products against cutaneous and possibly other forms of anthrax once screening tests are completed. PMID- 12549965 TI - Short-term oral cobalamin therapy for food-related cobalamin malabsorption. PMID- 12549966 TI - Commonly overlooked sources of vitamin K. PMID- 12549967 TI - Ballism associated with bupropion use. PMID- 12549968 TI - Slowing the progression of renal disease in patients with diabetes mellitus. PMID- 12549969 TI - Comment: hospital admissions resulting from preventable adverse drug reactions. PMID- 12549975 TI - Role of nitric oxide and oxidative stress in baroreceptor dysfunction in patients with chronic heart failure. AB - Abnormalities of autonomic control of the cardiovascular system are seen in chronic heart failure (CHF) and confer a poor prognosis. Nitric oxide appears to be important in the regulation of baroreflex control in health and in disease states. The antioxidant vitamin C increases nitric oxide bioavailability in CHF. We evaluated the effects of vitamin C on baroreceptor sensitivity (BRS) by sequence analysis in 100 CHF patients and 44 control subjects. Groups of 55 CHF patients and 22 controls were randomly allocated to receive a single intravenous injection of vitamin C (2 g) or placebo. In addition, 45 CHF patients were randomly allocated to receive a 4-week course of oral vitamin C (4 g/day) or placebo. An age-related reference range for BRS was developed in 22 healthy controls matched for age and gender to the CHF group. BRS was significantly impaired in the CHF group compared with age-matched older controls and young controls (6.9 +/- 3.1, 12.5 +/- 4.9 and 21.7 +/- 9.1 mmHg/ms respectively; P < 0.001 between groups). Intravenous vitamin C acutely improved BRS in CHF patients by 24% (by 1.8 +/- 4.1 mmHg/ms; P < 0.05), but not in controls. There was no improvement in BRS in CHF patients given chronic oral vitamin C. Thus acute intravenous, but not chronic oral, vitamin C improved BRS in CHF patients. There was no effect of intravenous vitamin C in healthy subjects, suggesting that the mechanism was either by free radical scavenging or due to central effects. PMID- 12549976 TI - Modulation of neutrophil complement receptor 3 expression by pneumococci. AB - Complement receptor 3 (CR3; CD18/CD11b) plays an important role in the recognition and clearance of Streptococcus pneumoniae (pneumococci) by neutrophils. The purpose of the present study was to characterize the modulation of CR3 surface expression on neutrophils exposed to pneumococci and to assess its functional significance. CR3 was detected with fluorescent phytoerythrin-labelled anti-CR3 (CD11b) antibodies, quantified with a fluorescence cell counter (FACS) and localized by confocal fluorescence microscopy. Uptake of fluorescent FITC labelled pneumococci was quantified by FACS. Whole blood from healthy volunteers was exposed at 37 degrees C to killed whole type III Streptococcus pneumoniae (KSP; 10(8)/ml) or to a positive control ( Escherichia coli lipopolysaccharide) that enhanced CR3 surface expression on neutrophils to a comparable extent. Varying the concentration of KSP between 10(5) and 10(8) organisms/ml progressively augmented CR3 surface expression measured at 1 h, whereas the response declined at 10(9)/ml. The diminished response to 10(9) KSP/ml proved to be time-dependent, with surface CR3 up-regulated maximally within 5 min, and down regulated thereafter. Labelling of CR3 during exposure demonstrated accelerated receptor sequestration, and confocal fluorescence microscopy demonstrated internalized CR3. Cooling to 16 degrees C, to inhibit the up-regulation of CR3 surface expression, also inhibited the uptake of FITC-labelled KSP and morphological changes. Accelerated down-regulation of surface CR3 expression by exposure to 10(9)/ml unlabelled KSP diminished the uptake of labelled KSP added subsequently. In contrast, lipopolysaccharide-induced up-regulation of CR3 expression increased the uptake of labelled KSP. Together, these experiments reveal dynamic modulation of CR3 expression on the surface of neutrophils exposed to pneumococci and a functional correlate of this modulation. Thus neutrophil expression of CR3 changes dynamically in response to exposure of neutrophils to progressively higher concentrations of pneumococci, conditions that mimic early neutrophil recruitment to densely infected lung tissue in acute pneumococcal pneumonia. PMID- 12549977 TI - Chaperone properties of Escherichia coli thioredoxin and thioredoxin reductase. AB - Thioredoxin, thioredoxin reductase and NADPH form the thioredoxin system and are the major cellular protein disulphide reductase. We report here that Escherichia coli thioredoxin and thioredoxin reductase interact with unfolded and denatured proteins, in a manner similar to that of molecular chaperones that are involved in protein folding and protein renaturation after stress. Thioredoxin and/or thioredoxin reductase promote the functional folding of citrate synthase and alpha-glucosidase after urea denaturation. They also promote the functional folding of the bacterial galactose receptor, a protein without any cysteines. Furthermore, redox cycling of thioredoxin/thioredoxin reductase in the presence of NADPH and cystine stimulates the renaturation of the galactose receptor, suggesting that the thioredoxin system functions like a redox-powered chaperone machine. Thioredoxin reductase prevents the aggregation of citrate synthase under heat-shock conditions. It forms complexes that are more stable than those formed by thioredoxin with several unfolded proteins such as reduced carboxymethyl alpha lactalbumin and unfolded bovine pancreatic trypsin inhibitor. These results suggest that the thioredoxin system, in addition to its protein disulphide isomerase activity possesses chaperone-like properties, and that its thioredoxin reductase component plays a major role in this function. PMID- 12549978 TI - Differential effects of hyperphosphorylation on splicing factor SRp55. AB - Members of the serine/arginine-rich (SR) protein family play an important role in both constitutive and regulated splicing of precursor mRNAs. Phosphorylation of the arginine/serine dipeptide-rich domain (RS domain) can modulate the activity and the subcellular localization of SR proteins. However, whether the SR protein family members are individually regulated and how this is achieved remain unclear. In this report we show that 5,6-dichloro-1 beta-D-ribofuranosyl benzimidazole (DRB), an inhibitor of RNA polymerase II-dependent transcription, specifically induced hyperphosphorylation of SRp55 but not that of any other SR proteins tested. Hyperphosphorylation of SRp55 occurs at the RS domain and appears to require the RNA-binding activity. Upon DRB treatment, hyperphosphorylated SRp55 relocates to enlarged nuclear speckles. Intriguingly, SRp55 is specifically targeted for degradation by the proteasome upon overexpression of the SR protein kinase Clk/Sty. Although a destabilization signal is mapped within the C-terminal 43-amino acid segment of SRp55, its adjacent lysine/serine-rich RS domain is nevertheless critical for the Clk/Sty mediated degradation. We report for the first time that SRp55 can be hyperphosphorylated under different circumstances whereby its fate is differentially influenced. PMID- 12549979 TI - The identification and characterization of human Sister-of-Mammalian Grainyhead (SOM) expands the grainyhead-like family of developmental transcription factors. AB - The Drosophila gene grainyhead is the founding member of a large family of genes encoding developmental transcription factors that are highly conserved from fly to human. The family consists of two main branches, with grainyhead as the ancestral gene for one branch and the recently cloned Drosophila CP2 as the ancestral gene for the other. We now extend this family with the identification of another novel mammalian member, Sister-of-Mammalian Grainyhead (SOM), which is phylogenetically aligned with grainyhead. SOM is closely related to the other mammalian homologues of grainyhead, including Mammalian Grainyhead (MGR) and Brother-of-MGR, sharing a high degree of sequence identity with these factors in the functional DNA-binding, protein dimerization and activation domains. Protein interaction studies demonstrate that SOM can heterodimerize with MGR and Brother of-MGR, but not with the more distant members of the family. Like grainyhead, the SOM gene too produces several distinct isoforms with differing functional properties through alternative splicing. The tissue distributions of these isoforms differ and all display highly restricted expression patterns. These findings indicate that SOM, like its family members, may play important roles in mammalian development. PMID- 12549980 TI - A rapid and sensitive fluorometric assay method for the determination of nitrilase activity. AB - A rapid, simple and sensitive fluorometric assay method for the determination of nitrilase activity is described. 3-Cyanopyridine was hydrolysed to nicotinic acid by Rhodococcus rhodochrous and the liberated NH(3) was allowed to react with buffered o -phthaldialdehyde-2-mercaptoethanol solution (pH 7.4) to form a fluorochrome. The fluorescence intensity was found to be stable after 20 min incubation at room temperature, and the optimum pH for the reaction was found to be 7.4. The fluorescence intensity was linearly related to enzyme activity with the substrate concentration ranging from 100 to 1000 mM. The activity determined by the proposed method correlates ( r =0.9625) well with the established Berthelot method. The proposed method is more sensitive than the existing methods for the determination of nitrilase activity. PMID- 12549981 TI - Computer aided analysis of additional chromosome aberrations in Philadelphia chromosome positive acute lymphoblastic leukaemia using a simplified computer readable cytogenetic notation. AB - BACKGROUND: The analysis of complex cytogenetic databases of distinct leukaemia entities may help to detect rare recurring chromosome aberrations, minimal common regions of gains and losses, and also hot spots of genomic rearrangements. The patterns of the karyotype alterations may provide insights into the genetic pathways of disease progression. RESULTS: We developed a simplified computer readable cytogenetic notation (SCCN) by which chromosome findings are normalised at a resolution of 400 bands. Lost or gained chromosomes or chromosome segments are specified in detail, and ranges of chromosome breakpoint assignments are recorded. Software modules were written to summarise the recorded chromosome changes with regard to the respective chromosome involvement. To assess the degree of karyotype alterations the ploidy levels and numbers of numerical and structural changes were recorded separately, and summarised in a complex karyotype aberration score (CKAS). The SCCN and CKAS were used to analyse the extend and the spectrum of additional chromosome aberrations in 94 patients with Philadelphia chromosome positive (Ph-positive) acute lymphoblastic leukemia (ALL) and secondary chromosome anomalies. Dosage changes of chromosomal material represented 92.1% of all additional events. Recurring regions of chromosome losses were identified. Structural rearrangements affecting (peri)centromeric chromosome regions were recorded in 24.6% of the cases. CONCLUSIONS: SCCN and CKAS provide unifying elements between karyotypes and computer processable data formats. They proved to be useful in the investigation of additional chromosome aberrations in Ph-positive ALL, and may represent a step towards full automation of the analysis of large and complex karyotype databases. PMID- 12549983 TI - hsCRP and HDL effects of statins trial (CHEST): rapid effect of statin therapy on C-reactive protein and high-density lipoprotein levels A clinical investigation. AB - Inflammation contributes to the pathogenesis of coronary heart disease and elevated serum levels of C-reactive protein (CRP) are independently associated with increased coronary risk. This study assessed whether there were differences in the effects on CRP and high-density lipoprotein (HDL) cholesterol levels among patients treated with three common statins. In a prospective, observational study, 80 dyslipidemic adults without evidence of cardiovascular disease were treated with 10 mg atorvastatin (A), 20 mg simvastatin (S), or 40 mg pravastatin (P) daily. CRP and lipid profiles were assayed before and after 12 weeks of therapy; in 21 patients, CRP levels were also measured after 1 and 4 weeks. The three treatment groups experienced comparable reductions in CRP (A: 33%, S: 42%, and P: 30%) and statistically insignificant changes in HDL cholesterol levels. CRP began to decrease after 1 week of treatment, and decreased further at 4 and 12 weeks of therapy. The change in the log-transformed CRP concentration correlated with the change in the log-transformed LDL cholesterol concentration. Subjects had similar baseline CRP levels, lipid profiles, and coronary risk factors. The authors conclude that at doses achieving similar reductions in LDL cholesterol, the three statins were associated with comparable decreases in CRP without significant changes in HDL cholesterol levels. The correlation between the reductions in CRP and LDL cholesterol differs from the findings of other published studies, and should prompt further investigation of the mechanism by which statins reduce CRP. PMID- 12549984 TI - Racial variations in cardiology care among hospitalized older heart failure patients. AB - In this retrospective follow-up study, the authors examined the association between race and the receipt of cardiology care in 1062 Medicare beneficiaries 65 years of age and older who were hospitalized with heart failure. The primary outcome measure was receipt of care from a cardiologist (via admission or consultation). Using logistic regression analyses, crude and adjusted odds ratios (OR) and 95% confidence intervals (95%CI) of receipt of cardiology care were estimated for nonwhite versus white patients. Two hundred (19%) patients were nonwhites and 483 (46%) patients received care from cardiologists. Proportion of patients receiving cardiology care was lower among nonwhite patients (35% versus 48% among whites; P = 0.001), and nonwhite race was associated with a lower odds of receiving cardiology care (crude OR = 0.57; 95%CI = 0.42-0.79). After adjustment for various patient characteristics and process-of-care variables, the magnitude and precision of the association between nonwhite race and a lower odds of receiving care from a cardiologist remained unchanged (adjusted OR = 0.43; 95% CI = 0.30-0.62). These findings suggest that nonwhite elderly hospitalized heart failure patients are less likely to be cared for by cardiologists. PMID- 12549985 TI - Cerebral hyperperfusion after angioplasty and stenting of a totally occluded left subclavian artery: a case report. AB - Cerebral hyperperfusion syndrome is documented after angioplasty of carotid and vertebral artery lesions. The authors report the first instance of cerebral hyperperfusion syndrome in the posterior cerebral circulation after angioplasty and stenting of a totally occluded left subclavian artery. PMID- 12549986 TI - Pharmacotherapy of diabetes mellitus: implications for the prevention and treatment of cardiovascular disease. AB - Diabetes mellitus in adults is associated with an increased risk of premature vascular disease and a higher mortality rate. The presence of other risk factors, often seen in diabetic patients, such as systemic hypertension, augments the rate of vascular diseases. Evidence is growing that tight control of hyperglycemia using insulin and/or oral hypoglycemic agents will modify this risk. More aggressive control of concomitant hypertension and/or hyperlipidemia is also required. Diabetic patients who have myocardial infarctions do worse than nondiabetic patients. Various strategies to improve outcomes include the use of tight blood glucose control, and various coronary interventions are currently under clinical study. PMID- 12549987 TI - Pharmacotherapeutic considerations in anesthesia. AB - This article focuses on new findings leading to improved understanding of the pathophysiology and mechanisms of potential drug interactions between anesthetic drugs or techniques and cardiovascular medications in patients scheduled for surgery. Only the most frequently used drugs are reviewed. Elective surgery provides the luxury to consider these risks and alter therapy accordingly. Under urgent circumstances, however, the increased risks associated with these agents should be anticipated with the goal to minimize adverse effects while maintaining optimal cardiovascular function in the perioperative period. PMID- 12549988 TI - Drug treatment of orthostatic hypotension and vasovagal syncope. AB - Orthostatic hypotension is a common problem, estimated to occur in 5 out of every 1000 individuals and in as many as 7% to 17% of patients in an acute care setting. Moreover, orthostatic hypotension may be more prominent in elderly patients due to the increased intake of vasoactive medications and concomitant decrease in physiologic function, such as baroreceptor sensitivity, often seen with aging. Orthostatic hypotension is a fall in blood pressure on assuming an upright position. Absolute cutoffs for the drop in blood pressure are often difficult to determine because different patients exhibit varying degrees of tolerance to falls in blood pressure. Therefore, strict numerical criteria may lead to underdiagnosis and improper intervention. A thorough review of patient symptomatology combined with appropriate clinical tests should be employed to narrow the vast differential diagnosis and pinpoint the etiology. The fall in blood pressure seen in orthostatic hypotension results from the inability of the autonomic nervous system to adequately compensate for the 500 mL blood that is estimated to pool in the lower extremities on assuming an upright posture. The decrease in venous return results in a concomitant decrease in cardiac output and thus hypoperfusion of the cerebral circulation, possibly resulting in syncope or various other symptoms. A complete investigation should consider hypovolemia, removal of offending medications, primary autonomic disorders, secondary autonomic disorders and, of course, vasovagal syncope, the most common cause of syncope. Although further research is still necessary to rectify the disease process responsible for orthostatic hypotension, patients suffering from this disorder can effectively be treated through a combination of nonpharmacologic treatment, pharmacologic treatment and patient education. Agents such as fludrocortisone, midodrine and erythropoietin show promising results as therapeutic adjuncts. Treatment for recurrent vasovagal syncope includes increased salt intake, and various drug treatments, most of which are still under investigation. PMID- 12549989 TI - Anemia and hypocholesterolemia. AB - The authors reviewed the available data on the relationship between hypocholesterolemia and anemia with various etiologies. The data were collected from sources identified by a MEDLINE search of English-language literature published between 1966 and 2001 and bibliographies of relevant articles. The study selection included primary research articles, metaanalyses, and abstracts concerning the relation between hypocholesterolemia and anemia. Textbooks, meeting proceedings, and reference lists were also searched. Despite differences in study design, a relation was found between hematocrit and serum cholesterol levels and between hemoglobin and cholesterol levels. Further studies are required to determine the exact etiology and clinical significance of these findings. PMID- 12549990 TI - Rosuvastatin: a new HMG-CoA reductase inhibitor for the treatment of hypercholesterolemia. AB - Because of their excellent tolerability and their positive impact on lipid parameters, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) have become the drugs of first choice for many patients with dyslipidemia. Rosuvastatin is an investigational statin in the U.S. with a number of favorable characteristics, which include low lipophilicity, high hepatocyte selectivity, minimal metabolism, and a low propensity for cytochrome P450 drug interactions. Rosuvastatin has been studied at doses ranging from 1 to 80 mg. In comparative clinical trials, rosuvastatin given at 5 to 10 mg/day reduced low density lipoprotein cholesterol to a significantly greater extent than atorvastatin 10 mg/day, pravastatin 20 mg/day, and simvastatin 20 mg/day. In addition, rosuvastatin exhibited beneficial effects on other lipid parameters such as high-density lipoprotein cholesterol and triglycerides. Rosuvastatin's safety profile was demonstrated to be similar to those of other statins. Given its favorable pharmacokinetic and pharmacodynamic characteristics, rosuvastatin is likely to become a valuable addition to the statin drug class. The author reviews the pharmacologic and pharmacokinetic properties of this new statin. PMID- 12549992 TI - [The current state of hormonal prevention of coronary heart disease in menopausal women]. AB - The increased incidence and prevalence of coronary heart disease among older women, coupled with the less favorable prognosis for women who sustain coronary events than for men, has resulted in the medical community's attention to the potential beneficial effects of hormone therapy in menopausal women. Much biological evidence supports a protective mechanism of estrogen; nevertheless, some aspects are contradictory. Although observational studies have shown a clear cardiovascular benefit associated with hormone therapy, the significant skew inherent in these data has resulted in overestimation of benefits and underestimation of risks. Recent reanalysis of these observational data controlling for confounding variables failed to show cardiovascular benefit. Several randomized, double-blind, placebo-controlled studies have failed to show improvement in clinical cardiovascular outcomes with menopausal hormone therapy both in healthy women and in women with established coronary heart disease. Current research has also focused on pharmacologic agents that selectively modulate estrogen receptors, such as raloxifene, which are useful for the prevention and treatment of osteoporosis without increasing the risk of breast cancer. A clinical trial is now underway to evaluate the effects of raloxifene on coronary events and on the incidence of invasive breast cancer in menopausal women both with established coronary heart disease and at increased risk for coronary events. Current recommendations do not advocate the initiation of menopausal hormone therapy for the primary or secondary prevention of coronary events. The proven lifestyle interventions of smoking cessation, heart healthy diet, weight control, and physical activity should be undertaken, with statin use for control of elevated LDL cholesterol levels and pharmacologic blood pressure control when appropriate. PMID- 12549993 TI - [Role of anti-inflammatory drugs in the treatment of acute coronary syndromes. From athero-inflammation to athero-thrombosis]. AB - Coronary thrombosis is the most important cause of morbidity and mortality and the most severe manifestation of atherosclerosis. Knowledge of the pathophysiology of atheroma formation and the causes of atheroma accidents have allowed the development of new therapeutic measures for reducing thrombotic events after a coronary episode. Treating the thrombosis after plaque rupture is useful, but a late measure once coronary flow is disturbed. Therefore, treatment at an earlier stage, which we call athero-inflammation, a central event in atheroma progression leading to atherothrombosis, seems wise. There is evidence of an inflammatory component in the pathogenesis of atheroma rupture in acute coronary events. Earlier studies of anti-inflammatory medication have not demonstrated a reduction in thrombotic complications after an acute coronary episode. However, there are pathophysiological arguments and clinical findings that suggest that it would be advisable to include anti-inflammatory medications, especially those that inhibit preferentially COX-2, in the therapeutic arsenal for this pathology. We postulated that blocking athero-inflammation could prevent thrombosis. A pilot study was carried out in 120 patients with acute coronary syndrome without ST-segment elevation in which 60 patients were treated with meloxicam, a preferential COX-2 inhibitor. All patients received heparin and aspirin. During the stay in the coronary care unit, as well as after 90 days, meloxicam lowered composite outcomes (myocardial infarction, death and revascularization procedures) compared with the control group. These results and available pathophysiological and clinical evidence support the hypothesis of potential benefits of non-steroidal anti-inflammatory drugs with preferential inhibitory activity on COX-2 in patients with acute coronary syndromes. More trials are needed to confirm their preventive effect. PMID- 12549994 TI - [Troponin and other markers of cardiac damage. Myths and realities]. PMID- 12549995 TI - [Heart failure: old questions, insufficient answers]. PMID- 12549996 TI - [Applicability of a new definition of myocardial infarction and the opinion of Spanish cardiologists]. AB - INTRODUCTION AND OBJECTIVES: The Working Group on Ischemic Heart Disease and Coronary Care Units of the Spanish Society of Cardiology evaluated the applicability of a new definition of infarction in Spanish hospitals, its current use, and the opinion of Spanish cardiologists. METHODS: A telephone survey was made (from late 2001 to early 2002) in Spanish hospitals to evaluate the availability of troponin or creatine kinase MB mass determinations. A questionnaire was sent to all members of the Spanish Society of Cardiology to query about the availability of determinations of cardiac necrosis markers at their respective hospitals, use of the new definition, and whether they agreed with the new definition. RESULTS: An important proportion of Spanish hospitals cannot determine myocardial necrosis markers (troponin or creatine kinase MB mass), mainly due to low-volume activity (fewer than 200 beds). The new definition of myocardial infarction was used by Spanish cardiologists always (24%), frequently (31%), sometimes (17%), seldom (14%), and never (11%). Agreement with the definition was complete in 21%, almost complete in 33%, half and half in 26%, rare in 10%, and absent in 7% of Spanish cardiologists. CONCLUSIONS: A large percentage of Spanish hospitals cannot use the new definition of myocardial infarction because they cannot determine specific cardiac necrosis markers. Spanish cardiologists are not generally using the new definition and many do not agree with it. PMID- 12549997 TI - [Long-term prognostic value of troponin I in patients admitted to a coronary unit for unstable angina]. AB - INTRODUCTION AND OBJECTIVES: Troponin I (TnI) is a useful marker of myocardial damage for the diagnosis and prognosis of acute coronary syndrome. The purpose of this study was to analyze the long-term prognostic value of the peak TnI concentration obtained within 48 h of admission to the coronary unit for unstable angina. METHODS: The study included 149 consecutive patients. Serial determinations were made of the MB fraction of creatine kinase (CK-MB) and TnI. Patients without CK-MB elevation were classified into two groups depending on the presence of high (n = 58) or normal (n = 91) troponin I values. We prospectively analyzed the clinical and evolutive factors related to the probability of death, new acute coronary event, or coronary revascularization at one-year of follow-up. RESULTS: There were no differences in the clinical characteristics between groups, except that patients in the group with high TnI values were older (69 vs. 64 years, p = 0.01). At one year of follow-up there were no differences in the incidence of new acute coronary events or coronary revascularization procedures; however there was a higher mortality in the group with high TnI (13 vs. 4%; p = 0.01). The independent predictors of mortality were prior myocardial infarction (RR = 3), elevated troponin I (RR = 3.2), left ventricular ejection fraction < 35% (RR = 10), and age > 70 years (RR = 15). CONCLUSIONS: In patients with unstable angina a high troponin I value in the first 48 h of admission was associated with a higher mortality rate at one-year of follow-up. PMID- 12549998 TI - [Prognostic value of troponin T in hospitalized patients with angina or non-ST segment elevation myocardial infarction]. AB - INTRODUCTION AND OBJECTIVES: Cardiac troponins are highly specific and sensitive for detecting minimal myocardial damage. The aim of our study was to determine the prognostic value of troponin T levels in patients hospitalized for suspected angina or myocardial infarction without ST-segment elevation. PATIENTS AND METHOD: We recorded the frequency of death, acute myocardial infarction, heart failure, or need for coronary revascularization in the three months after the onset of symptoms in 346 consecutive patients admitted for suspected acute coronary syndrome, excluding those who developed myocardial infarction with persistent ST-segment elevation. RESULT: . Serum troponin T levels were > or = 0.1 ng/ml in 133 patients (troponin T positive group) and lower in 213 patients (troponin T negative group). The relative risk (RR) and 95 percent confidence intervals (95% CI) of individual and grouped events for the troponin T positive group were 3.2 (95% CI, 1.4-7.3; p = 0.006) for death; 2.8 (95% CI, 1.43-5.51; p = 0.003) for death or myocardial infarction; and 2.8 (95% CI, 1.6-5.0; p < 0.001) for death, myocardial infarction or heart failure. Diabetes mellitus and troponin T levels > or = 0.1 ng/ml had independent prognostic value after adjusting for age, sex, and electrocardiographic changes; with RR 2.5 (95% CI, 1.01-5.9) for death, myocardial infarction or heart failure. CONCLUSIONS: The prognosis of patients hospitalized for chest pain who do not immediately develop transmural necrosis depends on serum troponin T levels at hospital admission. Troponin T levels > or = 0.1 ng/ml almost triple the risk of major events in the three months after the acute episode. The prognostic value of troponin T is independent of age, sex, presence of diabetes mellitus, and electrocardiographic changes. PMID- 12549999 TI - [Chest pain in clinical practice: impact of routine troponin determination on clinical manifestations and care]. AB - INTRODUCTION AND OBJECTIVES. To study the significance of chest pain in the clinical practice of a Spanish hospital and to evaluate the impact of routine troponin determination. METHODS: In our institution, routine serial measurements of troponins I and T were made in the evaluation of chest pain in 2000. We compared the results obtained in 1999 for all patients who visited the emergency room for chest pain and the patients who were hospitalized. We recorded the diagnosis at discharge, duration of the hospital stay, and associated costs. RESULTS: In 2000, 1,820 patients with chest pain visited the emergency department, which was equivalent to 1.9% of visits and 7.5 cases per 1,000 people and year: 43% of these patients were hospitalized for suspected acute coronary syndrome as compared to 49% in 1999 (-12%; p > 0.001). Among the patients admitted, 28% were discharged with a diagnosis of non-ischemic chest pain. Troponin determinations were associated with a lower probability of admission due to unstable angina (11.5 vs 16.0%; -28%; p < 0.001) and non-ischemic chest pain (12.1 vs 14.5%; -16%; p < 0.05), and an increase in diagnoses of non-Q wave acute myocardial infarction (3.4% vs 1.8%; +89%; p < 0.01). Non-ST elevation acute coronary syndrome ACS required 3,751 days of hospitalization and 1,003,420 euros of cost, and troponin determinations were associated with a reduction in hospital stays of 832 days (-18.2%) and 185,100 euros (-15.6%). CONCLUSION: Chest pain had a high incidence, 7.5, and generates high costs in hospital admissions. The routine use of serial troponin determinations was associated with a reduction in hospital admissions due to unstable angina and non-ischemic chest pain, and costs. PMID- 12550000 TI - [Use of diagnostic and therapeutic resources in patients hospitalized for heart failure: influence of admission ward type (INCARGAL Study)]. AB - BACKGROUND: Heart failure (HF) is the most rapidly growing cardiac pathology in industrialized countries, and already the primary cause of hospital admissions of elderly people. Outside the field of clinical trials, there have not been many studies in Spain of the influence of the admission department on diagnostic and therapeutic management, whether this affects short-term and long-term prognosis, and the factors that determine the department the patient is admitted to. OBJECTIVE: . To analyze whether management and prognosis of patients admitted with heart failure differ depending on the admission ward (cardiology versus internal medicine-geriatrics). PATIENTS AND METHODS: Cross-sectional study of 951 patients (505 men and 446 women) consecutively hospitalized for HF in the cardiology (n = 363) and internal medicine-geriatrics (n = 588) wards of 12 hospitals of Galicia and recruited over a maximum period of 6 months. The main epidemiological and clinical variables were recorded at admission, and the complications, treatments, and clinical status were recorded at release.Results. HF patients had a mean age of 75.5 12 years (women 78.5 years and men 72.6 years). The average hospitalization time was 11 8 days and 50.8% were first admissions. Total hospital mortality was 6.8%. Fifty-nine percent (58.9%) of patients had arterial hypertension, 31.9% ischemic heart disease, 27.6% cardiac valve disease, 28.5% diabetes mellitus, and 32.5% chronic obstructive pulmonary disease (COPD). The patients admitted to cardiology ward were younger (72.5 13 vs 77.4 11 years; p < 0.005), more frequently men (51.9 vs 43.7%; p < 0.005), more often first hospitalizations (54.8 vs 48.4%; p < 0.005), and acute pulmonary edema was more common (22.8 vs 9.2%; p < 0.005). The odds ratio (and 95% CI) for therapeutic and diagnostic procedures in relation to admission ward (reference group internal medicine-geriatrics), adjusted for age, sex, systolic function, number of hospitalizations, and history of dementia, hypertension, COPD, AMI, valve disease and ischemic heart disease, are: echocardiogram, 3.49 (2.58-4.73); catheterization, 6.42 (3.29-12.55), admission to intensive care, 3.94 (2.15 7.25), revascularization, 2.15 (0.57-8.08), and beta-blocker treatment, 3.39 (1.93-5.97). No differences in hospital mortality (6.6% in cardiology vs 7% in internal medicine-geriatrics) or average hospitalization time were found between departments. CONCLUSIONS: The admission ward was related with a clear difference in HF management, with better adherence to guidelines and more use of resources by cardiologists. This was unrelated with differences in hospital mortality so a longer follow-up of these patients is required to evaluate the impact of these therapeutic measures on the prognosis and evolution of HF, as well as the cost benefit relation in an elderly patient population. PMID- 12550001 TI - [Exercise echocardiography to differentiate dilated cardiomyopathy from ischemic left ventricular dysfunction]. AB - OBJECTIVES: Previous studies have shown the usefulness of dobutamine echocardiography to differentiate dilated cardiomyopathy (DC) from ischemic left ventricular dysfunction (ILVD), but no studies have been made using exercise echocardiography (EE). We hypothesized that most patients with DC have some contractile reserve and experience an increase in left ventricular ejection fraction (LVEF) during exercise, as opposed to patients with ILVD. Differences in response to EE may be useful to clinically differentiate between these two entities. PATIENTS AND METHOD: Between 1 March 1995 and 1 March 2001, we performed 4,133 EE studies on 3,830 patients. Of 289 patients (8%) with moderate or severe LV dysfunction (biplane LVEF < 41% and left ventricular end-diastolic diameter > 5.2 cm), 207 were excluded: 111 for a history of myocardial infarction; 28 for scarring on echocardiography (regional akinesia/dyskinesia with thinning and/or increased brightness); 13 for previous revascularization procedures; 9 for aortic valve disease; 11 for a known cause of cardiomyopathy; and 35 for not undergoing angiography. The study group was therefore composed of 82 patients who were encouraged to perform maximal treadmill EE. EE criteria for ILVD were either impaired regional wall motion (RWM) or a decrease/no change in LVEF from baseline to peak exercise, while criteria for DC were RWM improvement/no change and LVEF increase. The ILVD group was formed by 39 patients with stenosis >/= 70% diameter stenosis of a major epicardial coronary artery or major branch vessel. The remaining 43 patients constituted the DC group. RESULTS: The number of coronary risk factors (ILVD 2.0 1.1; DC 1.9 1.1), baseline LVEF (ILVD 30 7; DC 30 8), and exercise-induced angina (ILVD 23%; DC 14%) did not differ between groups (p = NS). ILVD patients achieved less Mets (6.6 3.1 vs 8.3 2.8; p < 0.05), had a lower heart rate x systolic blood pressure product (22 5 vs 27 7; p < 0.001), and developed regional and/or global LV dysfunction more frequently (79 vs 28%; p < 0.001). Sensitivity, specificity, positive and negative predictive values and global accuracy for ILVD detection were 79% (95% CI: 70-88), 72% (95% CI: 63-81), 72% (95% CI: 63-81), 79% (95% CI: 67-85), and 76% (95% CI: 69-83), respectively. CONCLUSION: Global and/or regional LV function impairment with exercise is accurate in identifying patients with ILVD. This method could reduce the need for invasive procedures. PMID- 12550002 TI - [Identification of factors responsible for oral over-anticoagulation in outpatients with heart disease]. AB - BACKGROUND: Few studies have attempted to investigate the clinical course or identify factors responsible for excessive anticoagulation in patients with heart disease. OBJECTIVES: To determine the incidence of excessive anticoagulation in outpatients with heart disease treated with acenocoumarol, analyze the factors related with over-anticoagulation, and identify bleeding complications. PATIENTS AND METHOD: This 7-month prospective observational study included consecutive outpatients anticoagulated with acenocoumarol. They were seen in an anticoagulation unit. The high INR group of 55 over-anticoagulated patients had at least one test with INR > 5. The control group of 49 patients had INR results strictly within therapeutic range. RESULTS: A total of 3,683 INR determinations were made in 512 patients. Seventy-seven tests had an INR > 5 (a 2% overall incidence of high-INR). In the group of 55 INR < 5 patients, 31% had more than one INR determination > 5 during follow-up. Multivariate analysis identified four variables as independent predictors of over-anticoagulation: artificial heart valve, poor treatment compliance, addition of potentially interactive new drugs, and illness in the last month. The high-INR group patients had more bleeding episodes (21.8 vs 4.08%; p = 0.008), one of which was major. CONCLUSION: The incidence of excessive oral anticoagulation in our outpatient population was similar to that reported in other studies. Patients with INR > 5 had more total bleeding complications, mostly minor. It is recommended to proceed carefully with oral anticoagulant therapy in patients with an artificial heart valve, suspected poor treatment compliance, addition of potentially interactive new drugs, and illness in the last month. PMID- 12550003 TI - [Congenital heart disease in adults: residua, sequelae, and complications of cardiac defects repaired at an early age]. AB - Nowadays, it is estimated that 85% of the infants born with congenital heart disease (CHD) will survive to adulthood, thanks mainly to surgical or therapeutic procedures performed during infancy or childhood. The clinical profile and disease pattern of adults with CHD is changing. The prevalence of certain adult CHDs, such as tetralogy of Fallot, transposition of the great arteries or univentricular heart, is rising, but these conditions have practically become new diseases as a result of therapy. Most surviving patients present residua, sequelae, or complications, which can progress during adult life. These disorders can present electrophysiological disturbances, valvular disease, persistent shunts, myocardial dysfunction, pulmonary or systemic vascular disease, problems caused by prosthetic materials, infectious complications, thromboembolic events, or extravascular disorders involving multiple organs or systems. In tetralogy of Fallot, the most striking problems that affect long-term prognosis are pulmonary valve regurgitation, right ventricle dysfunction, and atrial or ventricular arrhythmias. The main problems appearing after physiological atrial repair of transposition of the great arteries are related to right ventricular function, since it is structurally unprepared for systemic circulation, and atrial arrhythmias. Surgical repair of univentricular heart using Fontan techniques should be considered a palliative procedure that does not modify the underlying structural disorder and exposes the postoperative patient to severe complications and problems. The increase in the number of patients with CHD who will reach adulthood in the coming decades makes it necessary to carefully consider the new healthcare demands that are being generated, who should be responsible for them, and how and where solutions can be found. PMID- 12550005 TI - [Subtotal stenosis of a single right coronary ostium]. PMID- 12550006 TI - [Assessment of three activity questionnaires in patients with heart failure]. AB - The activity questionnaire most frequently used in heart failure, the New York Heart Association Classification, does not correlate well with peak oxygen uptake. The correlation of this variable with other activity questionnaires was analyzed in 83 patients (69 with heart failure and 14 control patients), 61.5 11 years old, who were interviewed and classified according to the Canadian Cardiovascular Society Classification. The Dyspnea-Fatigue Index and functional capacity according to the Specific Activity Questionnaire were determined for each patient. Subsequently, the treadmill cardiopulmonary exercise test (Naughton) was performed and the following correlations with peak oxygen uptake were found: Canadian Cardiovascular Society, r = 0.39; Dyspnea-Fatigue Index, r = 0.44; Specific Activity Questionnaire, r = 0.38. p < 0.001 for all three. The Dyspnea-Fatigue Index yielded the best correlation, although it was only slightly better than the Canadian Cardiovascular Society Classification, which is easier to obtain. PMID- 12550007 TI - [Transhepatic access for radiofrequency ablation in a child]. AB - Radiofrequency catheter ablation is safe and effective in children with refractory supraventricular tachycardia and poor response to pharmacological treatment. This procedure may be difficult or impossible to perform in patients with bilateral obstruction of the femoral venous access. In recent years, the transhepatic approach has been found to be safe in children. We report the case of a 19-month-old boy with permanent junctional reciprocating tachycardia who underwent transhepatic catheterization for radiofrequency ablation. PMID- 12550008 TI - [Normal conventional electrocardiogram with negative pharmacological stress test does not rule out Brugada syndrome]. AB - The diagnosis of Brugada syndrome, or right bundle-branch block with an elevated ST segment and negative T waves in V1-3, is obscured by the transitory normalization of the electrocardiogram, which can be unmasked by administering sodium-channel blockers. It has been recently reported that the condition can be underdiagnosed if only conventional precordial leads are used. We present the cases of two asymptomatic patients, a mother and son, with a family history of sudden cardiac death in first-degree relatives. Baseline ECGs obtained in conventional leads and one and two intercostal spaces above conventional electrode sites were similar, normal in the mother and saddle-like in the son. A flecainide stress test elicited the characteristic pattern of Brugada syndrome in both patients, but only in the high leads. Pharmacological stress testing with conventional precordial lead recordings does not rule out Brugada syndrome. We recommend that ECG recordings should include leads in the second and third intercostal spaces. PMID- 12550009 TI - [Training in radiological protection for interventional cardiology]. PMID- 12550011 TI - [Clinical practice guidelines and the practice of medicine]. PMID- 12550012 TI - [Immigration and tuberculosis]. PMID- 12550013 TI - [Screening program for alpha-1 antitrypsin deficiency in patients with chronic obstructive pulmonary disease, using dried blood spots on filter paper]. AB - Alpha-1 antitrypsin (AAT) deficiency is an under-diagnosed disease and screening programs have therefore been recommended for patients with chronic obstructive pulmonary disease (COPD). We present the results of the pilot phase of a screening program for AAT deficiency in order to evaluate the technique used, the procedures for transporting samples and the results obtained. Over a period of one month, five centers collected samples from all COPD patients for whom plasma concentrations of AAT or Pi phenotype had not yet been determined. Capillary blood spots were dried on filter paper and then sent by surface mail to a central laboratory for study. An immunonephelometric assay was used to determine AAT and DNA phenotyping was done by use of a Light Cycler. Samples were analyzed from 86 COPD patients (76 men, 10 women) with a mean age of 68.2 years. AAT deficiency was ruled out for 74 patients (86%) who had concentrations above the cutoff established, although one of them was MZ heterozygote by genotype. Among the 12 remaining patients (13.9%), only two also had a Z allele. The rest were individuals with concentrations below the established threshold and no evidence of a Z allele (10 patients, 11.6%). The Z allele frequency observed (3/172; 1.74%) was very similar to that found in the general population. The results of this pilot study allowed us to confirm that the method used to collect samples worked well. The sampling method is applicable, easy and well-accepted by participating physicians. It allowed AAT concentrations and Z allele deficiency to be determined. The method correlates well with standard techniques used for samples in whole blood. PMID- 12550014 TI - [Non-invasive ventilation in kyphoscoliosis. A comparison of a volumetric ventilator and a BIPAP support pressure device]. AB - Non-invasive intermittent positive pressure ventilation (NIPPV) at home is the treatment of choice for patients with chronic respiratory insufficiency secondary to severe kyphoscoliosis. Our aim was to compare clinical course, blood gases and lung function after one month of domiciliary NIPPV with two types of ventilator and to assess sleep pattern changes in patients enrolled in a prospective, randomized crossover study. Ten patients with chronic respiratory insufficiency due to kyphoscoliosis were enrolled and randomly assigned to the first device. After one month of use, the patients underwent clinical and functional examinations and polysomnographic studies while using the ventilator. The same protocol was applied with the second device after a ten-day washout period. Baseline polysomnographs showed fragmented sleep with low percentages of deep non REM sleep and of REM sleep, as well as respiratory patterns characterized by very high frequencies coinciding with significant desaturations. In all cases symptoms and arterial blood gas improvements were significant, with no differences between the two treatment periods. The percentages of time spent with SaO2 below 90% of reference in sleep studies were significantly lower than baseline with both ventilators. All but one patient had better tolerance of the bilevel positive airway pressure (BIPAP) support mode than of the volumetric ventilator. Our study shows that NIPPV is equally effective for patients with kyphoscoliosis whether administered with a volumetric ventilator or a BIPAP device. Subjective response and tolerance seem to be slightly better with BIPAP. PMID- 12550015 TI - [Sympathetic dermatomes corresponding to T2 and T3 ganglia. A prospective study of 100 superior thoracic sympathicolytic procedures]. AB - The autonomic nervous system adjusts sweating on the body's skin surface. The skin areas corresponding to the various sympathetic ganglia are not yet clearly defined. The purpose of this study was to define the skin areas corresponding to the T2 and T3 sympathetic ganglia. METHODS AND MATERIALS: We performed bilateral thoracic sympathicolysis endoscopically on the T2 and T3 ganglia of 50 patients with primary hyperhidrosis. All answered a questionnaire before surgery and at least two months later. The questionnaire asked about sweating at different points, but in order to estimate the cutaneous projection of T2 and T3 we only analyzed anhidrotic regions. RESULTS: Denervation resulted in anhidrotic hands (98.11%); axillas (73.58%); arms (39.62%); forehead (20.75%); chin, cheeks, neck and the nape of the neck (16.98%); sides of the chest (13.2%); front of the chest (7.54%); and back (1.88%). CONCLUSIONS: The territory influenced by the T2 and T3 sympathetic ganglia is more extensive than has been described by classical anatomical studies. The cutaneous area innervated by the sympathetic T2 and T3 ganglia extends to zones influenced by T1 in up to 20.75% of patients with primary hyperhidrosis. PMID- 12550016 TI - [Evaluation of clinical diagnostic guidelines for idiopathic pulmonary fibrosis]. AB - OBJECTIVES: To analyze the international consensus statement on diagnostic criteria for idiopathic pulmonary fibrosis. METHODS: All patients diagnosed of any interstitial lung disease by means of open lung biopsy since 1980 were included. The patients' clinical records were examined to determine whether they fulfilled the diagnostic criteria, and their biopsies were reviewed to find those with the usual interstitial pneumonia pattern. We calculated sensitivity, specificity, positive and negative predictive values and likelihood ratios for the diagnostic criteria in the consensus statement. Afterwards, we performed the analyses again using only one of the two conditions for fulfilling the function criterion. RESULTS: Of 39 patients enrolled in the study, 17 had idiopathic pulmonary fibrosis. Specificity and positive predictive value were both 100%, but sensitivity was 41.2% and negative predictive value was 68.7%. The likelihood ratio for a negative result was 0.59. In the second analysis, sensitivity was 64.7% and negative predictive value was 78.5%, while specificity and positive predictive value remained unchanged. The likelihood ratio for a negative result was 0.35. CONCLUSIONS: The criteria are sufficiently specific to allow us to diagnose confidently when the criteria are fulfilled. However, a slight change in the function criterion should be considered in order to increase diagnostic yield. PMID- 12550017 TI - [Mediastinoscopy in the diagnosis of mediastinal disease. An analysis of 181 explorations]. AB - To validate our experience with standard cervical mediastinoscopy (SCM) and extended cervical mediastinoscopy (ECM) to diagnose mediastinal nodes and masses, we studied 181 patients between January 1992 and February 2001. SCM and ECM were indicated for diagnostic staging of nodes related to bronchogenic carcinoma (Group I) or of mediastinal masses (Group II). An SCM was performed in all cases to explore the paratracheal region (2R, 2L, 4R, 4L, 7, 10R and 10L); in 21 additional cases, an ECM was performed to explore the aortopulmonary window or the subaortic region (area 5) and the para-aortic region (area 6). In Group I, the sensitivity of SCM was 93.6% and specificity was 100%; the positive predictive value (PPV) was 100%, the negative predictive value (NPV) was 82.8%, and the diagnostic yield was 95.1%. The sensitivity of ECM was 91% and specificity was 100%; PPV was 100%, NPV 93.3% and yield was 96%. In Group II, the sensitivity was 93.3%, specificity 100%, PPV 100%, NPV 81.2% and diagnostic yield 94.8%. The sensitivity of ECM in this group was 80%, specificity was 100%, PPV 100%, NPV 66.7% and yield 85.7%. A 2.7% complication rate was observed, with one case of bleeding after injury to the superior vena cava, one tracheal lesion, one recurring paralysis and two cases of surgical wound infection. The mean postoperative stay was 36 hours and mortality was zero. We conclude that SCM is highly specific for the evaluation of mediastinal node involvement in bronchogenic carcinoma and it is the approach of choice when a diagnosis of lesions located in the mid-mediastinal region has not been reached. ECM is a valid, safe alternative to anterior mediastinotomy for staging nodes and masses occupying para-aortic zones or the aortopulmonary window, with good diagnostic yield, low morbidity and absence of mortality. PMID- 12550018 TI - [Guidelines for a diagnostic and therapeutic approach to smoking addiction. A consensus report]. PMID- 12550019 TI - [Accreditation requirements for educational programs in respiratory medicine]. PMID- 12550020 TI - [Reference figures for pulmonary artery pressures after effective treatment of Graves' disease]. AB - We report the case of a 48-year-old woman with a diagnosis of pulmonary hypertension and hyperthyroidism (Graves' disease) in whom pulmonary artery pressures became normal after treatment of thyroid disease. The possible pathogenic mechanisms involved in this association include the presence of hyperdynamic heart failure and/or the presence of immune alterations underlying both conditions. PMID- 12550021 TI - [Is physiology destroyed?]. PMID- 12550023 TI - [Acute rhabdomyolysis in non-bacteremic pneumococcal pneumonia]. PMID- 12550024 TI - [Lag times between acceptance and publication of scientific articles. An analysis of editorial quality]. PMID- 12550025 TI - [Bone scintigraphy in oncology]. PMID- 12550026 TI - [Attenuation correction in studies of myocardial perfusion. When is it necessary?]. PMID- 12550027 TI - [Occult breast cancer. Detection and radioguided surgery with 99mTc-MIBI]. AB - We include those patients who present with an isolated metastasis of axillary adenopathy in the occult primary breast cancer group. Presumably, the primary tumor is a primitive breast carcinoma, unsuspected until this moment and not clinically demonstrable by mammography or ultrasonography. When no method succeeds in confirming the primary breast lesion, the patients are usually treated assuming the existing of breast cancer. Several diagnostic methods have been used to find the primary breast lesion. Magnetic Resonance imaging (MRI), Positron Emission (PET) and Doppler sonography have been used in this way and several papers present the results reached with them. Our group incorporates detection and radioguided surgery with 99mTechnetium (99mTc) methoxyisobutil isonitrile into the study of these patients. We perform a planar scintimammography and SPECT (Single Photon Emission Computed Tomography) with 99mTc-MIBI. If the radioisotopic method shows a functional image compatible with a carcinoma, a gamma detecting probe is then used to locate the lesion and guide its surgical removal. In this paper, we present the application of the technique in 5 cases and describe the technique and its possibilities. Its advantages are explained in comparison with other methods. The dosimetric values found in the performance of the technique are reported. We consider that detection and radioguided surgery with 99mTc-MIBI in the diagnosis and treatment of occult breast cancer adds an effective tool and means progress in the approach to this disease. PMID- 12550028 TI - [Diagnostic value of gated-SPECT for each coronary artery in patients clinically diagnosed of coronary heart disease]. AB - This study aimed to evaluate the diagnostic value of gated-SPECT for each coronary artery in patients with clinical diagnosis or suspicion of coronary heart disease. PATIENTS AND METHODS: The study population is made up of 43 patients (64 9 years, 88% male gender) with prior clinical diagnosis or suspicion of coronary heart disease who had undergone gated-SPECT (99mTc-tetrofosmin) and cardiac catheterization. Scintigraphic study after exercise treadmill test and rest study were performed on the same day. RESULTS: Gated-SPECT showed perfusion defects in 86% of patients, the mean number of territories with perfusion defects being 1.58 +/- 0.79. A total of 39 (91%) of the 43 patients had significant coronary heart disease. Single, two- and three-vessel disease was demonstrated in 12 (28%), 15 (35%) and 12 (28%) patients, respectively. Sensitivity and specificity were 81% and 91%, respectively, for left anterior descending artery, 88% and 65% for right coronary artery, and 55% and 81% for circumflex one. CONCLUSIONS: Myocardial scintigraphy with gated-SPECT offers high sensitivity and specificity for the diagnosis of left anterior descending artery disease. However, sensitivity for circumflex artery and specificity for right coronary artery were low in our series. PMID- 12550029 TI - [Clinical predictors of the presence of perfusion defects in more than than one territory in myocardial scintigraphy with GATED-SPECT]. AB - BACKGROUND AND OBJECTIVE: Extension of perfusion defects is associated with outcome in patients undergoing myocardial scintigraphy. The study aimed to identify the clinical characteristics that can predict the existence of perfusion defects in more than one territory in patients referred for myocardial scintigraphy with GATED-SPECT. PATIENTS AND METHODS: A total of 193 patients undergoing myocardial scintigraphy with GATED-SPECT (99mTc-tetrofosmine) were studied. Clinical variables and scintigraphy results were studied to determine what clinical variables are associated with perfusion defects in more than one territory. RESULTS: The number of territories with perfusion defects per patient was 1.1 0.8 and 29% had perfusion defects in > 1 territory. Patients with greater probability of having perfusion defects in > 1 territory were those with previous myocardial infarction (44% vs 21%, p = 0.030) and males (33% vs 10%, p = 0.006). In addition, patients with > 2 coronary risk factors had a statistical tendency to have defects in > 1 territory (47% vs 20%, p = 0.057). Only 15% of the patients with one of these three characteristics had perfusion defects in > 1 territory in comparison with 45% and 83% in those with 2 or 3 factors, respectively. CONCLUSION: Considering 3 simple clinical characteristics (male gender, previous infarction and existence of > 2 coronary risk factors), it is possible to predict which patients are more likely to show perfusion defects in > 1 territory during GATED-SPECT myocardial scintigraphy. PMID- 12550031 TI - [Value of 99mTc-Sulphur colloid scintigraphy in the diagnosis of intermittent digestive bleeding secondary to a case of jejunal angiodysplasia]. AB - We present the case of a 74 year old female patient, with clinical criteria of liver cirrhosis caused by hepatic C virus who required several admissions in our hospital over a five month period. She was given several blood transfusions because of repeated clinical manifestation of gastrointestinal bleeding without a clear origin. Her last admission was due to intermittent melenas, secondary anemia and hemodynamic angina. The patient's study included clinical analysis, fibrogastroscopy, colonoscopy, opaque enema, spiral CT and supraortic vessels arteriography. Because all the results to diagnose and locate the patient's disease were negative, she was referred to our service for a scintigraphy study. As it was an emergency case because of the patient's serious hemodynamic condition, a 99mTc-sulphur colloid scintigraphy was chosen, the results of which showed and located active gastrointestinal bleeding requiring urgent surgical intervention. The laparotomy with intrasurgical enterotomy and fibrogastroscopy undertaken ratified gastrointestinal bleeding, and the result of the biopsy of the jejunum removed in the intervention confirmed bowel angiodysplasia. Given the low incidence of gastrointestinal bleeding secondary to bowel angiodysplasia and absence of bibliographic references regarding the diagnosis of this disease in the above mentioned site by means of 99mTc-sulphur colloid scintigraphy in particular, we consider it interesting to highlight this case, in which the capability of this nuclear medicine technique for quick and non-invasive detection and location of gastrointestinal bleeding has been highly proven. PMID- 12550030 TI - [Evaluation of ventricular synchronization by fourier phase analysis in a radionuclide ventriculography]. AB - INTRODUCTION: Radionuclide ventriculography (RNV) evaluates segmental and global ventricular contractility and also detects conduction abnormalities. OBJECTIVE: To assess the temporal parameters of ventricular synchronization in the normal heart by a third harmonic (3H) Fourier phase analysis in a RNV and introduce this technique in our center. MATERIAL AND METHODS: Thirty normal subjects (19 men and 11 women) were included. An equilibrium RNV was performed in 35 degree left anterior oblique projection with 10 degree caudal tilt. The onset (T0); mean time (T(m)); total contraction time (T(t)); final time (T(f)) and propagation time (T(p)) for right (RV) and left ventricle (LV); as well as total propagation time (T(TP)); interventricular time (T(RV-LV)) and septum-lateral wall conduction time (T(S-LW)) were measured on the 3H Fourier histogram of the time-activity curve. RESULTS: Right ventricle contraction started 5 ms before that of the left ventricle (T(0RV) = 66 +/- 38 ms; T(OLV) = 71 +/- 30 ms), with a longer total contraction time (T(tVD) = 67 +/- 28 ms vs T(tVI) = 64 +/- 38 ms). Total propagation time (T(TP)) was 69 +/- 37 ms and the interventricular time (T(RV LV)) was 2 +/- 25 ms. Contraction progressed from septum to lateral wall, with a septum-lateral wall conduction time (T(S-LW)) of 4 +/- 22 ms. CONCLUSION: Simultaneous contraction of right and left ventricles can be quantified by RNV phase analysis, providing a useful tool for ventricular resynchronization assessment in multisite pacing. PMID- 12550033 TI - [Study of epidermal growth factor receptor concentration (EGFR) in lung adenocarcinomas. Association with high erbB2 oncogenic protein levels]. PMID- 12550032 TI - [Benign myocardial uptake of 99mTc-HMDP in prostate carcinoma: based on three cases]. AB - We present three cases of patients with prostate carcinoma who underwent total body bone scintigraphy with 99mTc-Hydroxymethylene diphosphonate (99mTc-HMDP), showing myocardial uptake. Complementary diagnostic examinations were performed in these patients: cardiac (electrocardiogram and echocardiography), analysis of plasma electrolytes, including calcium, review of personal background of each patient to rule out associated disease which could have produced this uptake. No evidence of associated disease was found, processes such as heart disease, amyloidosis, hyperparathyroidism, previous chemotherapy or radiotherapy being rule out. Thus, we conclude that the myocardial uptake was secondary to the prostate carcinoma. PMID- 12550034 TI - [Statistical parametric mapping (SPM) in nuclear medicine]. AB - In the scope of medical research, functional neuroimaging analysis permits the study of pathological or cognitive cerebral processes by using statistical quantification techniques. A tool of increasing use is the SPM (Statistical Parametric Mapping) software due to its wide availability and the variety of statistical studies that can be made. Nevertheless, being unaware of the theoretical background on which it is based may easily lead to inaccurate results and even to the reaching of erroneous conclusions. The present article summarizes these theoretical principles and discusses the main key points of the method without requiring advanced mathematical knowledge. PMID- 12550035 TI - [Economic studies in imaging diagnosis]. PMID- 12550036 TI - [Report from the editorial board]. PMID- 12550037 TI - [Betalactam resistant Streptococcus pneumoniae and management of pneumococcal meningitis]. PMID- 12550038 TI - [Epidemiologic study of infection by resistant gram-positive bacteria (G + R Study)]. AB - INTRODUCTION: This epidemiologic study was performed to obtain the following information: incidence of the main multiresistant gram-positive microorganisms (coagulase-negative staphylococci, Staphylococcus aureus, Streptococcus pneumoniae and enterococci) in Spanish hospitals, the type of infections they cause and the most highly affected hospital areas, as well as some antimicrobial resistance patterns, specifically S. aureus to methicillin and S. pneumoniae to penicillin. METHODS. Among the various Spanish autonomous regions, 104 hospitals and 446 doctors belonging to several medical fields participated in the study. Patients admitted to the following departments were included: Intensive Care, Infectious Diseases, Internal Medicine, Hematology, Surgery, Pneumology, Oncology and Pediatrics. The study was performed between December 2000 and March 2001, using two one-week time periods. RESULTS: Staphylococci were the most frequently isolated microorganisms, comprising more than 70% of the etiologic agents found. Enterococci were responsible for 17 to 20% of the infections evaluated. Among the total, 35.3% of S. aureus were methicillin-resistant and 28.6% of pneumococci were penicillin-resistant. The most frequent infections were bacteremias, followed by skin and soft tissue infections, and lower respiratory tract infections. CONCLUSIONS: This study confirms the high percentage of methicillin resistant S. aureus and penicillin-resistant S. pneumoniae present in our country. It also shows that among gram-positive microorganisms, there is a clear predominance of staphylococci as pathogenic agents. PMID- 12550039 TI - [Antibiotic resistance in 622 Streptococcus pneumoniae isolated from blood and cerebrospinal fluid in 33 Spanish hospitals participating in the European Antimicrobial Resistance Surveillance System (2000)]. AB - INTRODUCTION: In 1998 the European Union founded the European Antimicrobial Resistance Surveillance System (EARSS) for the control of antimicrobial resistance in invasive pathogens. This report present the results obtained for S. pneumoniae in the year 2000 in Spain. METHODS: Participants included 33 hospitals covering approximately 25% of the Spanish population. The laboratories carried out isolation, identification and sensitivity studies with their usual methods. Quality control was performed by the UK National External Quality Assurance Scheme (NEQAS). A form was filled out with clinical information on the patient and data on the hospital, department and specimen isolated. This information was entered in a central database for analysis and validation by the Whonet.5 program. RESULTS: Invasive S. pneumoniae was isolated in 622 patients. Overall incidence was 6.4 cases/100,000 inhabitants. Among the strains isolated, 33.1% (CI 95%: 29.6-37) showed resistance to penicillin (22.2% intermediate; 10.9% resistant); 11% showed resistance to cefotaxime (10.1% intermediate; 0.9% resistant); and 21.1% were resistant to erythromycin. Erythromycin resistance was 7% in penicillin-sensitive strains and 46.4% in penicillin-resistant strains (p < 0.001). A total of 51.7% of strains were resistant to penicillin in children < or = 4 years old, as compared to 29% in patients > 4 years old (p < 0.001). A ciprofloxacin MIC > 2 mg/L was found for 2.4% of isolates. CONCLUSIONS: Penicillin resistance in invasive S. pneumoniae in Spain is among the highest in the European Union. The decrease in sensitivity to penicillin was significantly associated with cefotaxime and erythromycin resistance. Children < or = 4 years old have a significantly higher risk than adults of acquiring infection with strains resistant to penicillin, cefotaxime and erythromycin. PMID- 12550040 TI - [Q fever in Gran Canaria: 40 new cases]. AB - INTRODUCTION: The aim of this study was to describe the clinical and epidemiologic features of Q fever in the southern area of the island of Gran Canaria (Spain). METHODS: We conducted a retrospective analysis of the clinical and epidemiological data of the cases of Q fever diagnosed by the Microbiology Laboratory of the Hospital Universitario Insular in Gran Canaria between 1998 and 2000. Antibodies against phase II Coxiella burnetii antigens were detected using an indirect immunofluorescence test. The diagnosis of acute Q fever was established by IgG titers > or = 1:320 and IgM titers > or = 1:80, or by seroconversion. RESULTS: During the period of study 59 cases of acute Q fever were diagnosed, making an incidence of 5 cases/100,000 inhabitants/year. The seroprevalence (IgG > or = 1:80) in the patients for whom Q fever serology was requested during that period was 23.9%. Clinical and epidemiologic data were available for 40 patients. All were sporadic cases and 57% were hospitalized. The mean age of the patients was 40.6 6 13.3 years (range 20-74 years), 85% were males and 67.5% came from a rural background. The majority of cases (65%) clustered from April to July. The most frequent clinical presentation was an acute febrile process with elevated liver enzymes (87.5%). Pneumonia was infrequent (only three cases). CONCLUSIONS: In our area Q fever is mainly manifested as an acute febrile illness with subclinical hepatic involvement. This fact and the small number of cases with pneumonia and chronic forms suggest the etiological involvement of C. burnetii strains different from those in other geographic areas. PMID- 12550041 TI - [Incidence of bacteremia among histologic groups of hematologic malignancies]. AB - INTRODUCTION: Subgroups having dissimilar prognoses are being identified among cancer patients with infection. Previous studies have suggested that these differences may be related to the histologic diagnosis, but this issue has not as yet been demonstrated. METHODS: We reviewed the medical records of all patients admitted with acute leukemia (AL) or lymphoma (ML) from 1988 to 1998. Incidence of bacteremia was calculated for the following subgroups: acute lymphocytic leukemia (ALL), acute myelocytic leukemia (AML), AML following refractory anaemia with excess blasts (AML-RAEB), high-grade ML (HGML), intermediate-grade ML (IGML), low-grade ML (LGML) and indeterminate ML (IML). Kaplan-Meier curves of time to the first positive blood culture were constructed and compared by means of log-rank test. RESULTS: In the period covered there were 244 new diagnoses of AL or ML: 62 AML, 32 ALL, 20 AML-RAEB, 78 HGML, 7 IGML, 37 LGML and 6 IML. At the end of the study period, 44 patients were alive, 147 were known to have died at a certain date and 53 had been formally lost to follow-up (most of them, transferred for hospice care). Among 684 blood cultures, there were 51 contaminations and 155 significant isolates. Among the latter, gram-positive bacteria were isolated in 74 and gram-negative bacteria in 47; in 27 cases more than 1 bacterial species were recovered. Fungi were isolated in 7 cases. The incidence of bacteremia expressed as cases per 1000 patient-days was 5.80 for AML, 5.03 for AML-RAEB, 1.56 for ALL, 0.21 for HGML and 0.40 for the remaining ML. Time to the first positive blood culture was significantly shorter for AML than for any other group, and was shorted for ALL and AML-RAEB than for ML. CONCLUSION: Differences in the incidence of bacteremia were observed among histologically-defined groups of unselected patients with hematologic malignancies. PMID- 12550042 TI - [Role of linezolid in antimicrobial therapy]. AB - The progressive emergence of multi-resistant gram-positive strains has prompted the search for new molecules (quinolones, streptogramins, oxazolidinones, ketolides, glycopeptides, daptomycin) to add to the current therapeutic arsenal. Linezolid, the first commercially available member of the oxazolidinone family, has evidenced activity against multi-resistant gram-positive strains (methicillin resistant Staphylococcus aureus, S. aureus with decreased glycopeptide sensitivity, vancomycin-resistant Enterococcus spp., Streptococcus pneumoniae with decreased sensitivity to penicillin and cephalosporins), thereby providing a new option for treating infections by these microorganisms. This work reviews the microbiologic and pharmacologic aspects of this agent in order to establish its position among the available options for antimicrobial chemotherapy. PMID- 12550043 TI - [beta-Lactam antibiotics]. AB - Betalactams, which act by inhibiting the last phase of bacterial cell wall synthesis, constitute the largest family of antimicrobial agents and the most extensively used in current clinical practice. These drug have a slow bactericidal action that is relatively independent of plasma concentrations, little toxicity and a broad therapeutic margin. Their spectrum has increased over the years with the incorporation of new molecules having greater activity against gram-negative bacilli. Nevertheless, the progressive emergence of acquired resistance has limited the empirical use of betalactams and their efficacy in certain situations. Despite this problem, penicillin is still the treatment of choice for a large number of classic infections, cephalosporins are widely used in surgical prophylaxis and severe community-acquired infections, carbapenems are the choice for mixed nosocomial and multiresistant bacterial infections and betalactamase inhibitors permit the effective use of amino- and ureido penicillins in highly significant infections. PMID- 12550044 TI - [Male with fever and left hypochondrial pain of several weeks duration]. PMID- 12550045 TI - [Unusual finding in peripheral blood]. PMID- 12550046 TI - [SEIMC protocol on osteoarticular and soft tissue infection, and asymptomatic bacteriuria: value of extended prophylaxis]. PMID- 12550048 TI - [Role of demand in the prescription of antibiotics]. PMID- 12550049 TI - [Joint prosthesis infections: several comments]. PMID- 12550051 TI - [Pseudozyma sp.: colonizer of the respiratory tract in critical patients]. PMID- 12550052 TI - [Tetanus: a historical curiosity?]. PMID- 12550054 TI - Oxidized products of cholesterol: dietary and metabolic origin, and proatherosclerotic effects (review). AB - Cholesterol oxidation products, termed oxysterols, are increasingly considered of potential interest in the pathogenesis of atherosclerotic lesions. Of dietary or endogenous origin, oxysterols may occur in significant amounts in low density lipoprotein (LDL) particles, especially in hypercholesterolemic subjects. They likely contribute to the uptake of modified LDL by scavenger receptors and some of them finally accumulate in the subintimal space of major arteries; here cholesterol oxides may favor the perpetuation of a chronic inflammatory state, through their ability to trigger irreversible damage of vascular cells with consequent activation of phagocytes. Furthermore, practically all oxysterols of major pathophysiologic interest have been shown to markedly up-regulate expression and synthesis of adhesion molecules, inflammatory cytokines and chemokines. Cholesterol oxidation thus appears to be an important biochemical pathway through which it exerts toxic, inflammatory and finally atherogenic effects. PMID- 12550055 TI - Fatty acid modulation of MCF-7 human breast cancer cell proliferation, apoptosis and differentiation. AB - Epidemiological studies suggest that dietary polyunsaturated fatty acids (PUFA) may influence breast cancer progression and prognosis. In order to study potential mechanisms of action of fatty acid modulation of tumor growth, we studied, in vitro, the influence of n-3 and n-6 fatty acids on proliferation, cell cycle, differentiation and apoptosis of MCF-7 human breast cancer cells. Both eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) inhibited the MCF 7 cell growth by 30% and 54%, respectively, while linoleic acid (LA) had no effect and arachidonic acid (AA) inhibited the cell growth by 30% (p < 0.05). The addition of vitamin E (10uM) to cancer cells slightly restored cell growth. The incubation of MCF-7 cells with PUFAs did not alter the cell cycle parameters or induce cell apoptosis. However, the growth inhibitory effects of EPA, DHA and AA were associated with cell differentiation as indicated by positive Oil-Red-O staining of the cells. Lipid droplet accumulation was increased by 65%, 30% and 15% in the presence of DHA, EPA and AA, respectively; (p < 0.05). These observations suggest that fatty acids may influence cellular processes at a molecular level, capable of modulating breast cancer cell growth. PMID- 12550056 TI - Chlorogenic acid modifies plasma and liver concentrations of: cholesterol, triacylglycerol, and minerals in (fa/fa) Zucker rats. AB - Chlorogenic acid, a phenolic compound found ubiquitously in plants, is an in vitro antioxidant and metal chelator. Some derivatives of chlorogenic acid are hypoglycemic agents and may affect lipid metabolism. Concentrations of cholesterol and triacylglycerols are of interest due to their association with diseases such as non-insulin-dependent-diabetes- mellitus and obese insulin resistance. As little is known about the effects of chlorogenic acid in vivo, studies using obese, hyperlipidemic, and insulin resistant (fa/fa) Zucker rats were conducted to test the effect of chlorogenic acid on fasting plasma glucose, plasma and liver triacylglycerols and cholesterol concentrations. Aditionally, the effects of chlorogenic acid on selected mineral concentrations in plasma, spleen, and liver were determined. Rats were implanted with jugular vein catheters. Chlorogenic acid was infused (5 mg/Kg body weight/day) for 3 weeks via intravenous infusion. Chlorogenic acid did not promote sustained hypoglycemia and significantly lowered the postprandial peak response to a glucose challenge when compared to the same group of rats before Chlorogenic acid treatment. In Chlorogenic acid-treated rats, fasting plasma cholesterol and triacylglycerols concentrations significantly decreased by 44% and 58% respectively, as did in liver triacylglycerols concentrations (24%). We did not find differences (p > 0.05) in adipose triacylglycerols concentration. Significant differences (p < 0.05) in the plasma, liver, and spleen concentration of selected minerals were found in chlorogenic acid-treated rats. In vivo, chlorogenic acid was found to improve glucose tolerance, decreased some plasma and liver lipids, and improve mineral pool distribution under the conditions of this study. PMID- 12550057 TI - Effect of cola intake on insulin resistance in moderate fat-fed weaning male rats. AB - In recent years, the prevalence of type 2 diabetes mellitus has dramatically increased in Korea as the diet has rapidly become westernized. We determined the effect of a long-term cola intake for insulin resistance in weaning male Sprague Dawley rats consuming a moderate fat diet. Thirty male pubs born from 6 female rats were randomized into cola or water drinking groups. The rats of the cola group were freely provided with 33 energy percent fat diets and cola for 28 weeks, while the rats of the control group had the same diet with water instead of cola. The daily caloric intake did not differ between groups, while the rats in the cola group consumed more carbohydrates. However, the mean body weight of the cola group was lower than that of the control group from the second week of the study. Whole body glucose disposal rates measured by euglycemic hyperinsulinemic clamp were higher in the cola group. Compared to the control group, glycogen contents and fraction velocity of glycogen synthase of the quadriceps muscle in the cola group were higher by 39.4% and 40.3%, respectively. Uncoupling protein (UCP)-2 and GLUT 4 contents of soleus and quadriceps muscles were higher in the cola group than the control group. In conclusion, insulin action improved with increased peripheral glucose utilization in weaning male rats drinking cola, which was partly due to lower body weight. This latter was possibly as a result of increased thermogenesis in muscles. PMID- 12550058 TI - The effect of dietary carbohydrate on the rise in plasma glutamate concentrations following oral glutamate ingestion in rats. AB - Plasma glutamate concentrations were examined in male rats following oral intubation of monosodium L-glutamic acid (MSG, 250 mg/kg) soon after ingesting one of several meals differing in carbohydrate content. Intubation of MSG alone produced a 4-fold rise in plasma glutamate that peaked at 15 min, and returned to baseline by 60 min. Red blood cell glutamate concentrations were unchanged. The ingestion of a meal lacking carbohydrate produced a modest attenuation of the post-MSG intubation rise in plasma glutamate concentrations. This attenuating effect increased progressively with the carbohydrate content of the meal (and as the protein content declined, to maintain isocaloric meals), though as little as 5% carbohydrate marked attenuated the plasma glutamate rise. This effect diminished as the time interval between the meal and MSG intubation increased from 1 to 4 hrs. Similar, but not identical effects were noted when meals substituted fat (instead of protein) for carbohydrate. The intubation of MSG alone produced a slight increase in plasma alanine concentrations over the 60-min post-intubation period examined. The ingestion of any of the meals just prior to intubation did not influence this effect. Overall, the results indicate that although the ingestion of carbohydrate can markedly attenuate the rise in plasma glutamate that follows MSG consumption in rats, this effect is also influenced by the other macronutrients present. The absence of notable, meal related changes in plasma alanine suggests that this parameter does not provide a useful indication of gut glutamate transamination. PMID- 12550059 TI - Exercise improves plasma lipid profiles and modifies lipoprotein composition in guinea pigs. AB - These studies were conducted to determine the effects of exercise training on plasma lipoprotein levels and metabolism in the guinea pig to evaluate potential utilization of this model for studies of exercise-mediated effects on the regulation of sterol and lipoprotein metabolism and atherosclerosis regression. Male guinea pigs (n = 5 per group) were randomly assigned to either a control or an exercise group. The exercise protocol consisted of a 7-week training program, 5 days/wk on a rodent treadmill. Final speed and duration were 33 meters/min for 30-40 min per session. Guinea pigs in the exercise group had 33% lower plasma triacylglycerol concentrations (P < 0.01), 66% higher HDL cholesterol levels (P < 0.05) and 31% lower plasma free fatty acids (P < 0.05) than guinea pigs from the non-exercised group. In addition, lipoprotein lipase activity in the heart was 50% higher (P < 0.025) in guinea pigs allocated to the exercise protocol. Exercise training resulted in modifications in composition and size of lipoproteins. The concentrations of free cholesterol in LDL and HDL were higher in the exercised guinea pigs. The LDL peak density values were lower in guinea pigs from the exercise group compared to controls suggesting that exercise training resulted in larger LDL particles. In contrast, no significant effects due to exercise were observed in hepatic cholesterol concentrations, hepatic HMG CoA reductase activity or LDL binding to guinea pig hepatic membranes. These data indicate that exercise had a more pronounced effect on the intravascular processing of lipoproteins than on hepatic cholesterol metabolism. In addition, the pattern of changes in guinea pig lipoprotein metabolism, in response to exercise training, was similar to reported effects in humans. PMID- 12550060 TI - Major phenolic compounds in olive oil: metabolism and health effects. AB - It has been postulated that the components in olive oil in the Mediterranean diet, a diet which is largely vegetarian in nature, can contribute to the lower incidence of coronary heart disease and prostate and colon cancers. The Mediterranean diet includes the consumption of large amounts of olive oil. Olive oil is a source of at least 30 phenolic compounds. The major phenolic compounds in olive oil are oleuropein, hydroxytyrosol and tyrosol. Recently there has been a surge in the number of publications that has investigated their biological properties. The phenolic compounds present in olive oil are strong antioxidants and radical scavengers. Olive "waste water" also possesses compounds which are strong antioxidant and radical scavengers. Typically, hydroxytyrosol is a superior antioxidant and radical scavenger to oleuropein and tyrosol. Hydroxytyrosol and oleuropein have antimicrobial activity against ATTC bacterial strains and clinical bacterial strains. Recent syntheses of labeled and unlabelled hydroxytyrosol coupled with superior analytical techniques have enabled its absorption and metabolism to be studied. It has recently been found that hydroxytyosol is renally excreted unchanged and as the following metabolites as its glucuronide conjugate, sulfate conjugate, homovanillic acid, homovanillic alcohol, 3,4-dihydroxyphenylacetic acid and 3,4-dihydroxyphenylacetaldehyde. Studies with tyrosol have shown that it is excreted unchanged and as its conjugates. This review summarizes the antioxidant abilities; the scavenging abilities and the biological fates of hydroxytyrosol, oleuropein and tyrosol which have been published in recent years. PMID- 12550061 TI - Cocoa procyanidin chain length does not determine ability to protect LDL from oxidation when monomer units are controlled. AB - Cocoa flavan-3-ols (catechin, epicatechin and oligomeric procyanidins) were tested for their ability to decrease LDL oxidative susceptibility and spare alpha tocopherol (alpha-toc) in vitro. Physiologic concentration (0.10-0.50 &mgr;M) of flavanols were used. The flavanols increased LDL conjugated diene lag times dose dependently from 23-207% and 15-143% in response to copper and AAPH oxidation, respectively, and delayed alpha-toc consumption. Sparing of LDL alpha-toc represents a possible mechanism for flavanols to enhance the resistance of plasma and LDL to oxidative stress. Procyanidins decreased LDL oxidative susceptibility with increasing chain length. However, when based on equivalent amounts of monomeric units, they inhibited LDL oxidation to a similar extent. This suggests that antioxidant activity of procyanidins with biologic substrates is not attributable to chain length or charge delocalization through polymeric linkages, but primarily to ring structures and catechol groups. Additionally, human plasma was analyzed for the presence of oligomeric procyanidins following consumption of a flavanol-rich cocoa product. Procyanidin dimers were detected in plasma concordant with the appearance of monomeric flavanols, with a peak of 0.08 +/- 0.01 &mgr;mol/L (n = 6) at two hours after consumption. Thus, this paper confirms the occurrence of procyanidins in human plasma, and extends previous structure function observations regarding flavanoid protection of LDL. PMID- 12550062 TI - Vitamin C and E supplements improve the impaired antioxidant status and decrease plasma lipid peroxides in hemodialysis patients small star, filled. AB - This study investigated the supplementation with vitamin C or/and E on the antioxidant system in hemodialysis patients. Thirty-eight hemodialysis patients (27 males and 11 females) with the average of 60 years old were divided into four groups: placebo (400 mg starch/time), vitamin C (400 mg/time)-, vitamin E (400 mg d,l- alpha-tocopheryl acetate/time)-, and vitamin C (400 mg/time) + E (400 mg d,l alpha-tocopheryl acetate/time)-supplemented groups for 6-week supplementation. The patients orally received three capsules of placebo or antioxidant(s) three times a week after finishing hemodialysis. Thirty-six healthy subjects (22 males and 14 females) with the average of 58 years old were recruited as the control group. Hemodialysis patients significantly decreased plasma vitamin C by 32%, erythrocyte glutathione by 26%, and plasma total antioxidant status by 9%, but increased plasma lipid peroxide levels by 102% compared with the control group at the baseline. The levels of plasma vitamin C and total antioxidant status significantly decreased by 24% and 18%, respectively, from the post-dialysate compared with those from the pre-dialysate. At week 6, vitamin C + E-supplemented group significantly increased plasma vitamin C and E, erythrocyte glutathione, and plasma antioxidant status, and inhibited plasma lipid peroxides compared with placebo group. Additionally, vitamin C + E-supplemented group had higher plasma vitamin C, vitamin E, and total antioxidant status, and lower plasma lipid peroxides than placebo group even at least 2 weeks after the termination of the supplements. Therefore, antioxidant vitamin supplements could improve antioxidant status and decrease lipid peroxides of hemodialysis patients. PMID- 12550063 TI - Hepatic reduction of insulin-like growth factor (IGF)-I and IGF binding protein-3 that results from fasting is not attenuated in genetically obese rats. AB - Fasting or caloric restriction causes substantial reductions in serum IGF-I in normal weight humans and animals, and reductions of liver IGF-I and IGFBP-3 mRNAs in animals. Obese humans, however, have attenuated and delayed decrements in IGF I in serum when subjected to caloric restriction. Obese Zucker rats show a clear tendency to preserve body protein during fasting. To determine whether obesity opposes the effects of fasting on IGF-I and IGFBP-3, and thereby contributes to preservation of lean tissue, we have examined the effect of 72 h of fasting on IGF-I and IGFBP-3 in lean and obese Zucker rats. We observe that between lean and obese animals, fasting for 72 h produces similar decrements in body weight, serum IGF-I, liver IGF-I mRNA, serum IGFBP-3 and liver IGFBP-3 mRNA. Our finding that the reduction of IGF-I and IGFBP-3 in liver that results from 72 h of fasting is not attenuated in obese Zucker rats raises the possibility that conservation of lean tissue in these animals during fasting is not related to the hepatic production of IGF-I and IGFBP-3. PMID- 12550064 TI - Role of soy isoflavones in the hypotriglyceridemic effect of soy protein in the rat. AB - The present study was undertaken to determine whether isoflavones present in soy protein isolate contribute to the triglyceride-lowering effect of the protein relative to casein. Plasma triglyceride concentrations, their secretion rate into blood circulation, and post-heparin plasma lipoprotein lipase activity (a major determinant of intravascular catabolism of triglycerides) were measured in the fasted state in male Sprague-Dawley rats fed for 21 days one of three experimental diets varying in protein source (20% weight/weight): soy protein isolate, casein or casein to which 1.82 mg/g isoflavones (genistein and daidzein) were added to match the isoflavone content of soy protein isolate. Body weight gain was slightly lower in soy protein fed rats than in casein fed rats, but this effect was not statistically significant (P = 0.22). Casein plus isoflavones diet induced intermediary weight gain. A decrease in plasma total triglycerides was observed in rats fed soy protein and casein plus isoflavones compared with casein (P < 0.05), and there was a tendency to a positive correlation between weight gain and plasma triglyceride concentrations (r = 0.35, P = 0.06). However, no significant effect was observed on hepatic triglyceride concentrations, triglyceride secretion rate by the liver and post-heparin plasma lipoprotein lipase activity. These results show that soy protein isolate, in comparison with casein, has a hypotriglyceridemic effect in the rat and suggest that isoflavones may be responsible, at least in part, for this effect. The lowering effect of soy protein isolate and isoflavones on plasma triglyceride concentrations may be mediated by an alteration in energy balance, and possibly by the hepatic production of lipoproteins more susceptible to intravascular hydrolysis. Subtle but sustained changes in triglyceride secretion and post-heparin plasma lipoprotein lipase activity may also be implicated. PMID- 12550065 TI - Dietary diacylglycerol suppresses high fat diet-induced hepatic fat accumulation and microsomal triacylglycerol transfer protein activity in rats. AB - We have recently shown that the long-term ingestion of dietary diacylglycerol (DAG) mainly containing 1,3-isoform reduces body fat accumulation in humans as compared to triacylglycerol (TAG) with the same fatty acid composition. The fat reduction in this human experiment was most pronounced in visceral fat and hepatic fat. Recent animal studies have also indicated that dietary DAG induces alteration of lipid metabolism in the rat liver. In the present study, the dietary effects of DAG on high fat diet-induced hepatic fat accumulation and hepatic microsomal triglyceride transfer protein (MTP) activity were examined in comparison with those of TAG diet in rats. When the TAG oil content was increased from 10 to 30 g/100 g diet, hepatic TAG concentration, hepatic MTP activity and MTP large subunit mRNA levels were significantly increased after 21 days. However, when the dietary TAG oil (30 g/100 g diet) was replaced with the same concentration of DAG oil with the same fatty acid composition, the increase of the TAG concentration and the MTP activity in the liver were significantly less and the mRNA levels remained unchanged. The MTP activity levels correlated significantly with hepatic TAG concentration.These results showed that dietary DAG may suppress high fat diet-induced MTP activity in the liver, and indicated the possibility that hepatic TAG concentration may regulate hepatic MTP activity. PMID- 12550066 TI - Differential effects of dietary flaxseed protein and soy protein on plasma triglyceride and uric acid levels in animal models. AB - The effect of dietary soy protein and flaxseed meal on metabolic parameters was studied in two animal models, F344 rats with normal lipid levels and obese SHR/N cp rats with elevated levels of cholesterol and triglyceride. The rats were fed AIN 93 diet differing only in the source of protein. The rats were fed either 20% casein, 20% soy protein or 20% flaxseed meal. Plasma was analyzed for cholesterol, triglyceride, uric acid, blood urea nitrogen (BUN), creatinine and total protein. In both strains of rats, flaxseed meal significantly decreased plasma cholesterol and triglyceride concentrations. The effect of soy protein on lipids was not as striking as that of flaxseed meal. Flaxseed meal also lowered uric acid in F344 rats and BUN in SHR/N-cp rats. Since cholesterol, triglyceride and uric acid are independent risk factors for cardiovascular disorders, our data show that both flaxseed meal and soy protein may have beneficial effects. Which chemical constituent(s) of flaxseed meal or soybean is (are) responsible for the beneficial effects need to be identified. PMID- 12550067 TI - Role of chromium supplementation in Indians with type 2 diabetes mellitus. AB - Type 2 diabetes mellitus is a complex metabolic disorder with adverse cardiovascular risk. The role of micronutrients has not yet been well clarified in this condition, especially in India.THE OBJECTIVES OF THIS STUDY WERE TO: (1) evaluate chromium status in Indian subjects with type 2 diabetes mellitus, (2) assess the effect of chromium picolinate (200 &mgr;g trivalent chromium twice daily) administration on glycaemic control and lipid profile in these subjects and (3) comment on the possible mechanism of any beneficial effect noted above.Fifty subjects were studied in a double blind, placebo-controlled, crossover fashion, with each treatment arm (chromium/placebo) lasting 12 weeks and 4 weeks' wash-off period in between. 50 healthy age- and sex-matched volunteers served as controls. Serum chromium level appeared to be higher in the general population in our country compared to western countries (36.5-59.5 nmol/L as compared to 2.3-40.3 nmol/L) However, the local diabetics were found to have a lower serum chromium level than the healthy controls (32.3 nmol/L against 44.7 nmol/L; p < 0.0001) and a mean increase of 3.5 nmol/L was noted after 12 weeks of chromium supplementation that was, expectedly, not seen in the placebo phase (p < 0.0001).Significant improvement in glycaemic control was noted in the chromium treated group (DeltaFasting serum glucose = 0.44 mmol/L, p < 0.001; DeltaPost prandial serum glucose = 1.97 mmol/L, p < 0.001; Deltaglycated hemoglobin = 0.01; p = 0.04, in comparison to placebo) This was accompanied by a significant greater fall in fasting serum insulin in the chromium-treated group, p < 0.05.The change in lipid parameters (total serum cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol and triglycerides) did not show significant difference between the chromium and placebo groups.Clinically significant hematological, renal or hepatic toxicity were excluded by routine hemogram, serum urea, creatinine, alanine amino transferase (ALT) and alkaline phosphatase estimations.In conclusion, chromium supplementation seems to improve glycaemic control in type 2 diabetic patients, which appears to be due to an increase in insulin action rather than stimulation of insulin secretion. PMID- 12550068 TI - Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships. AB - Flavonoids are a class of secondary plant phenolics with significant antioxidant and chelating properties. In the human diet, they are most concentrated in fruits, vegetables, wines, teas and cocoa. Their cardioprotective effects stem from the ability to inhibit lipid peroxidation, chelate redox-active metals, and attenuate other processes involving reactive oxygen species. Flavonoids occur in foods primarily as glycosides and polymers that are degraded to variable extents in the digestive tract. Although metabolism of these compounds remains elusive, enteric absorption occurs sufficiently to reduce plasma indices of oxidant status. The propensity of a flavonoid to inhibit free-radical mediated events is governed by its chemical structure. Since these compounds are based on the flavan nucleus, the number, positions, and types of substitutions influence radical scavenging and chelating activity. The diversity and multiple mechanisms of flavonoid action, together with the numerous methods of initiation, detection and measurement of oxidative processes in vitro and in vivo offer plausible explanations for existing discrepancies in structure-activity relationships. Despite some inconsistent lines of evidence, several structure-activity relationships are well established in vitro. Multiple hydroxyl groups confer upon the molecule substantial antioxidant, chelating and prooxidant activity. Methoxy groups introduce unfavorable steric effects and increase lipophilicity and membrane partitioning. A double bond and carbonyl function in the heterocycle or polymerization of the nuclear structure increases activity by affording a more stable flavonoid radical through conjugation and electron delocalization. Further investigation of the metabolism of these phytochemicals is justified to extend structure-activity relationships (SAR) to preventive and therapeutic nutritional strategies. PMID- 12550069 TI - Bioavailability of the isomer mixture of phytoene and phytofluene-rich alga Dunaliella bardawil in rat plasma and tissues. AB - Dunaliella bardawil, a beta-carotene-accumulating alga was treated by the bleaching herbicide norflurazon to select sub-species rich with a mixture of 9 cis and all-trans stereoisomers of phytoene and phytofluene. The present study determines the bioavailability of phytoene and phytofluene with their stereoisomers in rats fed on a diet supplemented with Dunaliella phytoene-rich spray dried powder. Three groups of female weanling rats, eight animals each, were fed AIN diets for two weeks. The control consumed the diet as is. The experimental group was supplemented with 50 g Dunaliella powder to give phytoene/phytofluene at a level of 1 g/kg diet, and the placebo was provided with the oxidized algae free of carotenoids at the same amount. Weight gain and tissues weight of rats fed on the control diet, or on the experimental diets were statistically same. Tissue analyses were carried out by liquid chromatography at the end of two weeks feeding for vitamin A, carotenoids, phytoene and phytofluene and theirs stereoisomers. Liver analyses revealed high hepatic storage of phytoene in the experimental group. Analysis of the other tissues, adrenal, brain, heart, kidney, lung, and spleen detected small amounts of phytoene in the adrenal, kidney and spleen and in the plasma. High-pressure liquid chromatography for stereoisomeric composition was performed to all phytoene-containing tissues. The original algal diet content of 9-cis-to-all-trans ratio of 1:1 was maintained in the plasma and adrenal while in the liver, spleen and kidney the ratio was reduced to 1:3. The preferential accumulation of all-trans phytoene over 9-cis phytoene in the liver, spleen and kidney may be interpreted as indicating stronger antioxidative effect of 9-cis phytoene over the all-trans isomer or alternatively, in vivo streoisomerization of 9-cis phytoene to the all-trans structure. PMID- 12550070 TI - Protective effect of polysaccharide from the loach on the in vitro and in vivo peroxidative damage of hepatocyte. AB - Misgurnus anguillicaudatus polysaccharide (MAP), a type of natural neutral polysaccharide occurring in the mucus of Misgurnus anguillicaudatus C., was studied for antioxidant bioactivity. The preliminary research showed that MAP was able to remove O(2)*, HO., H(2)O(2) and other active compounds of oxygen and significantly protected DNA chains from being damaged by hydroxyl radicals. This antioxidant bioactivity was further evaluated using the model of peroxide-induced oxidative injury in rat primary hepatocytes and hepatotoxity in mice. The results demonstrated that MAP, at the concentrations of 0.10 approximately 0.40 mg/ml, significantly increased the activity of SOD and GSH-px, but decreased the consumption of reduced glutathione and the formation of malondialdehyde induced by a 30-min treatment of H(2)O(2) (2.0 mmol/L). The in vivo investigation showed that oral pretreatment of MAP (100, 200 and 300 mg/kg) for 7 days before a single dose of t-BHP (0.2 mmol/kg, ip) significantly lowered the serum levels of hepatic enzyme markers (alanine and aspartate aminotransferase) and reduced oxidative liver injury. The histopathological evaluation of the liver revealed that MAP reduced the incidence of liver lesions including inflammatory, leukocyte infiltration, and necrosis induced by t-BHP in mice. Based on the results described above, a conclusion can be drown that MAP may play an important role in the prevention of oxidative damage in living systems. PMID- 12550071 TI - Assessment of the relationship between the antimutagenic action of riboflavin and glutathione and the levels of antioxidant enzymes. AB - In this study the role of antioxidant enzymes on the antimutagenic actions of riboflavin and reduced glutathione against mutagenic potentials of 4 nitroquinoline 1-oxide and mitomycin C have been investigated. For this purpose the activities of catalase and superoxide dismutase enzymes have been determined in Salmonella typhimurium TA102 and TA100 strains preincubated with different combinations of 4-nitroquinoline 1-oxide, mitomycin C, riboflavin and reduced glutathione for thirty minutes. Also in part of the same samples, the mutagenicity has been determined for each combination of chemicals by using Salmonella preincubation test. The correlation between the levels of antioxidant enzymes and mutagenicity and antimutagenicity has been investigated.While riboflavin displayed a weakly antimutagenic effect on 4-nitroquinoline 1-oxide mutagenicity in TA102 and TA100 (0.25, 0.35 inhibition respectively), it did not have any effect on the strong mutagenicity of mitomycin C in both strains. Reduced glutathione, a well known antioxidant, had no antimutagenic effect against the mutagenicity of both compounds in TA102 and TA100 strains. The antioxidant enzymes, catalase and superoxide dismutase, seemed to have no direct effect on the antimutagenic action of riboflavin and mutagenic action of 4 nitroquinoline 1-oxide and mitomycin C because no change in the activities of catalase and superoxide dismutase was detected in relation to antimutagenicity of riboflavin and mutagenicity of 4-nitroquinoline 1-oxide and mitomycin C in both strains. It should be noted that many antimutagens have more than one mechanism of action and their effect depends on the mutagens being tested. PMID- 12550072 TI - Comparative content of some bioactive compounds in apples, peaches and pears and their influence on lipids and antioxidant capacity in rats. AB - The aim of this study was to compare some bioactive compounds in apples, peaches and pears and their influence on lipids and antioxidant capacity in rats. The content of total polyphenols (g/100g) was 0.23 +/- 0.03; 0.22 +/- 0.03 and 0.68 +/- 0.1 in peeled fruits and 0.48 +/- 0.04, 0.47 +/- 0.04 and 1.2 +/- 0.12 in peels of peaches, pears and apples, respectively. Caffeic, p-coumaric and ferulic acids and the total radical-trapping antioxidative potential (TRAP) values in peeled apples and their peels were significantly higher than in peaches and pears, respectively. Contrarary, no significant differences in the content of dietary fiber among the studied fruits were found. The content of all studied indices in peels was significantly higher than peeled fruits (p < 0.05 ). A good correlation between the total polyphenols and the TRAP values was found in all fruits. Diets supplemented with apples and to a less extent with peaches and pears have improved lipid metabolism and increased the plasma antioxidant potential especially in rats fed with added cholesterol. The highest content of biologically active compounds and the best results in the experiment on rats makes apple preferable for dietary prevention of atherosclerosis and other diseases. PMID- 12550073 TI - Muscle and liver protein metabolism in rats fed raw or heat-treated pea seeds. AB - Raw or extruded pea (Pisum sativum, cv. Ballet) diets with or without supplementary amino acids were fed for 15 days to young growing rats and the effects on tissue weights, liver and muscle protein metabolism and hormone levels monitored. Body weight gain, liver and gastrocnemius muscle weights and protein contents were reduced and some key hormones altered when rats were fed unsupplemented raw pea diets. This appeared to be a result of amino acid deficiencies in the diet, the action of antinutritional factors and the refractory nature of the reserve proteins and other seed components. However, this did not in itself improve the nutritional performance of the rats due to the overriding effects of the amino acid deficiencies in the pea diets. After supplementation, extruded peas supported much higher rates of growth and skeletal muscle deposition than did supplemented raw peas. Despite this, the weight gains remained less than achieved on a high quality control diet. Protein synthesis and degradation rates in skeletal muscles and total protein contents were similar to control values. The lower growth rate did not appear to be due to impaired deposition of skeletal muscle. Deposition of other body components, possibly lipids, may have been lowered by supplemented extruded pea diets. Liver protein levels were reduced in rats fed supplemented raw peas and blood corticosterone was elevated. In conclusion, extrusion treatment of peas in combination with amino acid supplementation appeared to abolish the negative effects of peas on skeletal muscle deposition. PMID- 12550074 TI - Effect of DL-alpha-lipoic acid on the status of lipid peroxidation and protein oxidation in various brain regions of aged rats. AB - Free radicals have been implicated in the development of many acute and chronic diseases and in conditions involving brain or neurological tissue. The primary genetic material is subjected to damage by endogenous and exogenous agents, which may lead to instability and transcriptional infidelity. In the present study, we evaluated the protective effect of DL-alpha-lipoic acid, a metabolic antioxidant on lipid peroxidation, protein carbonyl content in various brain regions of aged rats when compared to brain regions of young rats. DL-alpha-lipoic acid was administered intraperitoneally (100mg/kg body weight/day) to experimental rats. Nucleic acid and protein content were low whereas thiobarbituric acid reactive substances and protein carbonyl content (markers of free radical damage) were high in cortex, striatum, hippocampus and hypothalamus followed by cerebellum of aged rat brain. Lipoate administration for 14 days in aged rats increased the levels of nucleic acid and protein and reduced lipid peroxidation and protein oxidation. These results demonstrate that lipoic acid is a potent antioxidant for neuronal cells against age associated oxidative damage. PMID- 12550075 TI - Dietary supplementation with cysteine prodrugs selectively restores tissue glutathione levels and redox status in protein-malnourished mice(1). AB - Protein malnutrition (PM) is a major health problem in the world. PM compromises antioxidant defense in the body. In particular, PM decreases tissue glutathione (GSH) levels. A high protein diet was found to restore tissue GSH levels in animal studies, however it is not recommended for the early phase of PM rehabilitation. Therefore, using dietary supplementation to restore tissue GSH without giving a high protein diet may be an adjunct therapy that helps improve antioxidant status during the early rehabilitation of PM. In this study, we systematically compared the efficacy of dietary supplementation of four cysteine prodrugs: N-acetylcysteine, L-2-oxo-4-thiazolidine-carboxylate, methionine, and GSH, on tissue GSH in mice fed a protein-deficient (0.5%) diet. Results showed that dietary supplementation of cysteine prodrugs to PM mice restored GSH levels in liver, lung, heart and spleen, but not in colon. GSH and GSSG levels in brain and kidney were not affected by cysteine prodrug or PM. Supplementation also restored the redox status in liver and heart (based on GSH/GSSG), and in liver and spleen (based on GSSG/2GSH reduction potential). This suggests that the restoration of GSH levels and redox status by cysteine prodrugs are tissue specific, and that the two indicators of redox status are not always interchangeable. However, all four prodrugs exhibited similar GSH-enhancing capacities, showing no prodrug-specificity as seen in cell culture studies. In conclusion, this study provided information that may be useful in a clinical setting where a short-term oral supplementation of cysteine prodrugs is necessary for the early rehabilitation of PM patients. PMID- 12550076 TI - The effect of treatment with the oral growth hormone (GH) secretagogue MK-677 on GH isoforms. AB - Growth hormone (GH) consists of several isoforms. We have studied the proportion, expressed as percentage of total GH concentration, of non-22kDa (non-22K) GH isoforms and 20K GH during 8-week oral treatment with MK-677 25mg daily in 12 obese males. The proportion of non-22K GH isoforms in peak total GH samples after the initial MK-677 administration was higher than that after 2 and 8 weeks (p<0.01 and p<0.05, respectively). In selected non-peak total GH samples after the initial MK-677 administration, however, the proportion of non-22K GH isoforms was similar to that in the peak total GH samples after 2 and 8 weeks. The proportion of 20K GH in 2-h samples after the initial MK-677 administration was lower than that after 2 and 8 weeks (p<0.01 and p<0.05, respectively). We concluded that the proportion of non-22K GH isoforms was higher in peak, but not in non-peak, total GH samples after the initial MK-677 administration than that observed after multiple doses. The proportion of 20K GH in 2-h samples after the initial MK-677 administration was lower than that after 2 and 8 weeks. These moderate changes in the proportion non-22K GH isoforms are likely of small importance for the clinical response to MK-677 treatment. PMID- 12550077 TI - Muscle development, insulin-like growth factor-I and myostatin mRNA levels in chickens selected for increased breast muscle yield. AB - Insulin-like growth factor-I (IGF-I) and myostatin (MSTN) are paracrine regulators of muscle growth. The present study was conducted to relate their expression with muscle fibre development in chickens selected for high breast meat yield and their controls. Both mRNA levels were measured by real-time RT-PCR in the Pectoralis major (PM) muscle between 14 days in ovo and 6 weeks post-hatch and in the Sartorius (SART) muscle between 2 and 6 weeks. The data show that PM growth was slow during in ovo development and rapid in the early post-hatch period. Chickens from the selected genotype exhibited significantly higher breast muscle yields from 2 to 6 weeks of age, and muscle fibre hypertrophy. In the PM, IGF-I and MSTN mRNA levels decreased markedly around hatch, while the IGF-I/MSTN ratio increased, suggesting that it could contribute to the explosive growth observed in the early post-hatch period. Between 4 and 6 weeks of age in selected chickens, IGF-I mRNA levels were significantly higher (p=0.04) with a similar trend in MSTN mRNA levels (p=0.07) in the PM muscle but not in the SART muscle. Our results support the hypothesis that the relative levels of IGF-I and MSTN mRNA may participate to set muscle growth rate along development, while other factors are required to explain differences between genotypes. PMID- 12550078 TI - Short- and long-term swimming exercise training increases myocardial insulin-like growth factor-I gene expression. AB - OBJECTIVES. We investigated the effect of short- and long-term swimming exercise, with or without insulin-like growth factor (IGF)-I administration, on the expression of myocardial IGFs and contractile proteins. METHODS: Sprague-Dawley male rats (n=36) were subjected to swimming exercise for 2 or 6 weeks. IGF-I (0.5mg/rat) was administered continuously for 1 week, using alzet osmotic pumps. Control groups remained sedentary. IGF-I, IGF-I receptor (IGF-IR), IGF-II, skeletal alpha-actin (sk-actin), and beta myosin heavy chain (beta MHC) mRNAs were measured using Northern blot analysis and RT-PCR. RESULTS: A significant 2 fold increase in myocardial IGF-I mRNA was found after 2 and 6 weeks of swimming in both IGF-I treated and untreated rats (p<0.001). IGF-IR mRNA was significantly (p<0.05) increased after 6 weeks of training only in the IGF-I treated animals. IGF-II mRNA remained unchanged at all time points. While beta MHC mRNA was significantly decreased (p=0.003) at 2 and 6 weeks, sk-actin mRNA remained unchanged. CONCLUSIONS: Short- and long-term swimming exercise training increase myocardial expression of IGF-I mRNA. Exogenous administration of IGF-I, during the first week of the exercise session, did not produce any effect on myocardial IGF-I but was associated with increased IGF-IR signal after the long-term exercise training. These data suggest a relationship between IGF-I expression and cardiac adaptation to exercise training. PMID- 12550079 TI - Growth impairment in IL-6-overexpressing transgenic mice is associated with induction of SOCS3 mRNA. AB - MUP/hIL-6 transgenic mice overexpressing human interleukin-6 (IL-6) are growth retarded. As documented here, the major transcriptional factor constitutively activated by IL-6 in the MUP/hIL6 transgenic mice was signal transducer and transactivator 3 (STAT3). Since STAT3 has been implicated in the expression of negative regulators of GH signaling, the suppressors of cytokine signaling (SOCS) genes, we have in this study examined the expression of SOCS1, SOCS2, SOCS3, and CIS genes. We found a large, 5-fold increase in SOCS3 mRNA in the liver, brain, skeletal muscle, and the lung of the MUP/hIL-6 transgenic mice. SOCS genes are thought to inhibit activation of transcriptional factor STAT5 by GH. Despite the induction of SOCS3 mRNA, STAT5 was activated in growth-retarded transgenic mice in response to elevated endogenous GH serum levels. The significance of activation of STAT3 and STAT5 transcription factors for cell proliferation and growth impairment in this mouse model is therefore discussed. PMID- 12550080 TI - Endoplasmic reticulum calcium storage and release in cells expressing misfolded growth hormone. AB - AIM: Deletion of amino acids 32-71 in human growth hormone (Delta 32-71-GH) causes severe autosomal dominant GH deficiency. These experiments test whether retention of Delta 32-71-GH in the endoplasmic reticulum (ER) lumen leads to aberrant Ca(2+) regulation. DESIGN: COS cells were transfected with Delta 32-71 GH, wild-type-GH or empty plasmid, and the ability of the cells to release Ca(2+) from the ER and transmit a Ca(2+) signal to the cytoplasm was investigated using cytoplasmic Ca(2+) dyes and ER-targeted cameleon Ca(2+) reporters. RESULTS: Resting free Ca(2+), the rate of Ca(2+) release from the ER and the size of the ionophore-releasable ER Ca(2+) pool were not altered by Delta 32-71-GH. Stimulation of endogenous Ca(2+)-mobilizing receptors for histamine and thrombin resulted in similar changes in cytoplasmic and ER Ca(2+) in cells expressing wild type and Delta 32-71-GH. CONCLUSION: Ca(2+) regulation is preserved despite retention of misfolded GH in the ER. PMID- 12550081 TI - The role of insulin-like growth factor-II in cancer growth and progression evidenced by the use of ribozymes and prostate cancer progression models. AB - Towards understanding the IGF system during cancer growth and progression, progressive prostate cancer models, such as SV40 large T antigen immortalized human prostate epithelial cells (P69, M2182, M2205, and M12) and LNCaP sublines (C4, C4-2, and C4-2B4), were used. IGF-II mRNA levels progressively increase as prostate cancer cells become more tumorigenic and metastatic, suggesting that IGF II contributes in part to prostate cancer progression. The role of IGF-II in cancer cell growth was evaluated in LNCaP, PC3, and M12 prostate cancer cell lines and MCF-7 breast cancer cell line by ribozyme/antisense strategies which were previously shown to suppress endogenous IGF-II expression and cell growth in PC-3 cells [Xu et al., Endocrinol 140 (1999) 2134]. Retroviral mediated transient expression of IGF-II-specific ribozyme (RZ) caused extensive cell death. In stably cloned cell lines, both RZ and mutant ribozyme (MRZ) inhibited cancer cell growth, suggesting that antisense effects of MRZ may be sufficient for cell growth inhibition. These results confirm an important role of IGF-II in cancer cell growth and progression, and support further development of gene therapy targeting IGF-II. PMID- 12550082 TI - Changes in summer temperature and heat-related mortality since 1971 in North Carolina, South Finland, and Southeast England. AB - Three climatically diverse regions were studied to determine the impact of temperature change on heat-related mortality from 1971 to 1997. Median regressions showed that May-August temperatures in North Carolina rose by 1.0 degrees C (95% CL 0.0-2.0 degrees C) from 23.5 degrees C (74.3 degrees F), were unchanged in South Finland at 13.5 degrees C (56.3 degrees F), and rose in Southeast England 2.1 degrees C (0.3-4.0 degrees C) from 14.9 degrees C (58.8 degrees F). After determining for each region the daily temperature (as a 3 degrees C band) at which the mortality was the lowest, annual heat-related mortality was obtained as excess mortality per million at temperatures above this. Annual heat-related mortality per million (among the population at risk, aged 55+) fell in North Carolina by 212 (59-365) from 228 (140-317) to only 16 (not significant, NS); fell in South Finland by 282 (66-500) from 382 (257-507) to 99 (NS); and fell in Southeast England by 2.4 (NS) from 111 (41-180) to 108 (41-176). The falls in North Carolina and South Finland remained significant after allowances were made for changes in age, sex, and baseline mortality. Increased air conditioning probably explains the virtual disappearance of heat related mortality in the hottest region, North Carolina, despite warmer summers. Other lifestyle changes associated with increasing prosperity probably explain the favorable trends in the cooler regions. PMID- 12550083 TI - Associations between ambient air pollution and daily mortality among persons with congestive heart failure. AB - We conducted a mortality time series study to investigate the association between daily mortality for congestive heart failure (CHF), and daily concentrations of particles and gaseous pollutants in the ambient air of Montreal, Quebec, during the period 1984-1993. In addition, using data from the universal Quebec Health Insurance Plan, we identified individuals >/=65 years of age who, one year before death, had a diagnosis of CHF. Fixed-site air pollution monitors in Montreal provided daily mean levels of pollutants. We regressed the logarithm of daily counts of mortality on the daily mean levels of each pollutant, after accounting for seasonal and subseasonal fluctuations in the mortality time series, non Poisson dispersion, weather variables, and other gaseous and particle pollutants. Using cause of death information, we did not find any associations between daily mortality for CHF and any air pollutants. The analyses of CHF defined from the medical record showed positive associations with coefficient of haze, the extinction coefficient, SO(2), and NO(2). For example, the mean percent increase in daily mortality for an increase in the coefficient of haze across the interquartile range was 4.32% (95% CI: 0.95-7.80%) and for NO(2) it was 4.08% (95% CI: 0.59-7.68%). These effects were generally higher in the warm season. PMID- 12550084 TI - Air pollution and emergency department visits for asthma among Ohio Medicaid recipients, 1991-1996. AB - We examined the effects of nitrogen dioxide (NO(2)), ozone (O(3)), particulate matter of <10 microm aerodynamic diameter (PM(10)), and sulfur dioxide (SO(2)) on asthmatics ages 5-34 years enrolled in Medicaid in Cincinnati, Cleveland, and Columbus, OH (N=5416). Our study period was for the summer months, June-August, from July 1, 1991 to June 30, 1996. We preformed Poisson regression analyses for the number of daily emergency department (ED) visits for asthma in each city and on the aggregate data controlling for time trends and minimum temperature. We found a 12% increased likelihood of an asthma ED visit per 50 microg/m(3) increase in PM(10) in Cleveland [95% confidence interval (CI)=0-27%] and a 35% increase per 50 microg/m(3) increase in SO(2) in Cincinnati (95% CI=9-21%). When data were analyzed for all three cities combined, the risk of an ED visit increased for all pollutant increases and specifically by 12% (95% CI=1-23%) per 50 microg/m(3) increase in SO(2). Attributable risk estimates show a five times greater impact on Cleveland over Cincinnati or Columbus. Between 1991 and 1996, air pollutants in Cincinnati, Cleveland, and Columbus increased ED visits for asthmatics enrolled in Medicaid. PMID- 12550085 TI - Arsenic in drinking water and adverse pregnancy outcome in an arseniasis-endemic area in northeastern Taiwan. AB - The well water in Lanyang Basin, which is located in the northeastern portion of Taiwan island, was found to have high levels of arsenic ranging from undetectable levels (<0.15 ppb) to 3.59 ppm. We performed a study to compare the risk of adverse pregnancy outcomes (preterm delivery and birthweight) between an area with historic high well water arsenic levels (arsenic-exposed area (AE)) and a comparison area with no historic evidence of arsenic water contamination (non arsenic-exposed area (NAE)). The mean birth weight in the AEs and NAEs were 3132.6 and 3162.6 g, respectively. Babies born in AEs were on average 30 g lighter than those born in NAEs. AEs had a higher rate of preterm delivery than NAEs (3.74% vs 3.43%). The results of this study suggest that, after adjustment for potential confounders, arsenic exposure from drinking well water was associated, although not significantly, with the risk of preterm delivery, with an odds ratio of 1.10 (0.91-1.33). The estimated reduction in birth weight was 29.05 g (95% CI=13.55-44.55). The findings from this investigation provide evidence for a potential role for arsenic exposure through drinking water in increasing the risk of low birthweight. PMID- 12550086 TI - Reactions of macrophages exposed to particles <10 microm. AB - This study describes experiments on cytotoxic effects and the production of oxidative radicals and the proinflammatory cytokine tumor growth factor alpha (TNFalpha) in a cell line of rat lung macrophages exposed to aqueous extracts from ambient air particles <10 microm (PM(10)) collected on Teflon filters. The particles were collected during the four seasons at two urban sites, one rural site, and one alpine site in Switzerland. Cytotoxic effects, determined as a reduction in the metabolic activity, were found in particle extracts from all sites and seasons. Taking together the data from all sites and seasons, a dose response function was observed between the particle mass on the filter and toxicity (r(2)=0.633, linear regression). The release of the pro-inflammatory cytokine TNFalpha as well as of oxidative radicals was most pronounced in particles collected in spring-summer and autumn. While at Montana (alpine), the stimulation of the cells was positively correlated with the particle mass on the filters, this correlation was negative at the urban sites Zurich and Lugano. It is interpreted that at high PM(10) levels, as in these cities, macrophages are inhibited by increasing air pollution due to toxic effects. Cytotoxic effects and the release of oxidative radicals could be inhibited when the extracts were treated with an endotoxin-neutralizing protein. This suggests that endotoxin, a cell-wall constituent of gram-negative bacteria, is one of the factors which modulates macrophage activity. All together, the experiments indicate that in the PM(10) fraction, water-soluble macrophage-toxic and macrophage-stimulating compounds are present. The data offer an explanation for at least some of the known harmful effects of PM(10), and confirm endotoxin as a possible reactant. PMID- 12550087 TI - Release of Streptomyces albus propagules from contaminated surfaces. AB - The release of Streptomyces albus propagules from contaminated agar and ceiling tile surfaces was studied under controlled environmental conditions in a newly developed aerosolization chamber. The experiments revealed that both spores and cell fragments can be simultaneously released from the colonized surface by relatively gentle air currents of 0.3 ms(-1). A 100 x increase of the air velocity can result in a 50-fold increase in the number of released propagules. The aerosolization rate depends strongly on the type and roughness of the contaminated surface. Up to 90% of available actinomycete propagules can become airborne during the first 10 min of the release process. Application of vibration to the surface did not reveal any influence on the aerosolization process of S. albus propagules under the tested conditions. This study has shown that propagules in the fine particle size range can be released in large amounts from contaminated surfaces. Measurement of the number of S. albus fragments in the vicinity of a contaminated area, as an alternative to conventional air or surface sampling, appears to be a promising approach for quantitative exposure assessment. PMID- 12550088 TI - The Comet assay for the detection of DNA damage in Mus spretus from Donana National Park. AB - Donana Park (Spain), a protected area in Europe, was affected by an environmental disaster in April 1998 that caused the spreading of acidic water and mud full of toxic metals from the Aznalcollar pyrite mine. In order to assess the contamination in the area and to monitor the possible biological effects of the toxic spill, a series of coordinated studies was performed utilizing several animal species living in that area. We performed genotoxicity monitoring using the Comet assay on peripheral blood leukocytes of the Algerian mouse (Mus spretus), a nonprotected rodent suitable as a bioindicator. The mice were sampled in different areas 6 months after the ecological disaster and again 1 year later. Our results showed that in 1998 all the areas examined were contaminated, as determined by an increase in the Comet assay parameters in the analyzed animals, whereas a significant decrease in the values of these parameters was observed in the 1999 samples, which were collected in a riverside area subject to tide flows. Thus, the Comet assay has proven to be an interesting and sensitive tool in studies of environmental genotoxicity. PMID- 12550090 TI - Lethal and subchronic effects of 2,4,6-trinitrotoluene (TNT) on Enchytraeus albidus in spiked artificial soil. AB - The effects of 2,4,6-trinitrotoluene (TNT) exposure in spiked artificial soil on the survival and reproduction rate of the white potworm Enchytraeus albidus were studied. Based on the initial concentrations, TNT in freshly spiked soil decreased enchytraeid survival (21-day LC(50)=422+/-63 (SD)mg/kg, N=3) and fecundity (42-day EC(50)=111+/-34, N=4). Data also indicated that TNT was 5-10 times more lethal to juveniles than adults, and lethality was less pronounced in TNT-spiked soils aged for 21 days. A time-dependent decrease in the TNT concentrations, as well as a concomitant increase in the levels of 2- and 4 aminodinitrotoluene, was observed during the 42-day toxicity test. Taken together, TNT (or one of its metabolites) is more lethal to juvenile than adult enchytraeids. This effect may explain, at least in part, the ability of TNT to decrease fecundity as determined using the enchytraeid mortality-reproduction test. PMID- 12550089 TI - Histopathology of the fish Corydoras paleatus contaminated with sublethal levels of organophosphorus in water and food. AB - The effects of contamination, through water or food, of a sublethal dose of the organophosphate methyl parathion were analyzed in tissues that are responsible for absorption (gills, intestine) and metabolism (liver), in the freshwater fish Corydoras paleatus. In gill respiratory lamellae, epithelial hyperplasia, edema, and detachment occurred, diminishing sooner after contamination by food than after contamination through water. In the intestine, lipoid vacuolization of enterocytes, apical cytoplasm, and an increase in goblet cell activity occurred mainly after ingestion of contaminated food. The liver exhibited cloudy swelling, bile stagnation, focal necrosis, atrophy, and vacuolization after contamination through both absorption routes, the highest degeneration being between T(8) and T(24). Metabolic processes that depend on liver function were equally impaired by the two routes of contamination, but secondary effects vary with gill and intestine pathologies as a consequence of water and food contamination, respectively. Therefore, a "safe" sublethal dose of methyl parathion causes serious health problems in C. paleatus. PMID- 12550091 TI - Water quality objectives for mixtures of toxic chemicals: problems and perspectives. AB - The need to develop water quality objectives not only for single substances but also for mixtures of chemicals seems evident. For that purpose, the conceptual basis could be the use of the two existing biometric models: concentration addition (CA) and independent action (IA), which is also called response addition. Both may allow calculation of the toxicity of mixtures of chemicals with similar modes of action (CA) or dissimilar modes of action (IA), respectively. The joint research project Prediction and Assessment of the Aquatic Toxicity of Mixtures of Chemicals (PREDICT) within the framework of the IVth Environment and Climate Programme of the European Commission, provided the opportunity to address (a) chemometric and QSAR criteria to classify substances as supposedly similarly or dissimilarly acting; (b) the predictive values of both models for the toxicity of mixtures at low, statistically nonsignificant effect concentrations of the individual components; and (c) the predictability of mixture toxicity at higher levels of biological complexity. In this article, the general outline, methodological approach, and some preliminary findings of PREDICT are presented. A procedure for classifying chemicals in relation to their structural and toxicological similarities has been developed. The predictive capabilities of CA and IA models have been demonstrated for single species and, to some extent, for multispecies testing. The role of very low effect concentrations in multiple mixtures has been evaluated. Problems and perspectives concerning the development of water quality objectives for mixtures are discussed. PMID- 12550092 TI - Dissipation of the herbicide oxyfluorfen in subtropical soils and its potential to contaminate groundwater. AB - The dissipation and mobility of the herbicide oxyfluorfen (2-chloro alpha,alpha,alpha-trifluoro-p-tolyl 3-ethoxy-4-nitrophenyl ether) in field soil of Taiwan were investigated in the laboratory with six tea garden soils. The dissipation coefficients of oxyfluorfen in soils of different moisture content (30%, 60%, and 90% of soil field capacity) and soil temperature (10 degrees C, 25 degrees C, and 40 degrees C) were studied. Results indicate that the half-life of oxyfluorfen ranged from 72 to 160 days for six tea garden soils. It was found that if the temperature is high, the dissipation rate is rapid, and there is almost no dissipation at 10 degrees C. Possible contamination of groundwater by the herbicide oxyfluorfen was assessed using the behavior assessment model and the groundwater pollution-potential (GWP) model. The results obtained after evaluating the residue and travel time using the GWP model illustrated that oxyfluorfen is not very mobile in soil and may not contaminate groundwater under normal conditions. But in the case of soil of extremely low organic carbon content and coarse texture, oxyfluorfen has the potential to contaminate groundwater less than 3m deep. PMID- 12550093 TI - 19F NMR spectroscopic investigation into the absorption and metabolism of 3 trifluoromethylaniline in Eisenia veneta. AB - 19F nuclear magnetic resonance (NMR) spectroscopy was used as a specific tool to investigate the metabolism of 3-trifluoromethylaniline (3-TFMA) in the earthworm species Eisenia veneta. Exposure was via a filter-paper contact toxicity test using five exposure levels (1000, 100, 10, 1, and 0.1 microg/cm(2)). Instant lethality was observed at the two highest levels. Worms exposed at the lower levels appeared to tolerate the compound. The 19F label of 3-TFMA allowed the uptake and metabolism of the earthworms to be monitored by 19F NMR spectroscopy. Metabolism of 3-TFMA was observed at 10 microg/cm(2) and, to a lesser extent, at 1 microg/cm(2). The possibility of 3-TFMA accumulation in specific organs was also investigated. As a simplified model, worms were cut into distinct anatomical regions (head, testes, crop, clitellum, and gut). At the two highest exposure levels, "uniform distribution" was observed. However, accumulation appeared to be proportional to the "size" of the extracted segments at the lower levels. PMID- 12550094 TI - Chronic toxicity of arsenic to the Great Salt Lake brine shrimp, Artemia franciscana. AB - We determined the chronic toxicity of arsenic (sodium arsenate) to the Great Salt Lake brine shrimp, Artemia franciscana. Chronic toxicity was determined by measuring the adverse effects of arsenic on brine shrimp growth, survival, and reproduction under intermittent flow-through conditions. The study commenced with <24-h-old nauplii, continued through reproduction of the parental generation, and ended after 28 days of exposure. The concentrations tested were 4, 8, 15, 31, and 56mg/L dissolved arsenic. The test was conducted using water from the Great Salt Lake, Utah as the dilution water. Adult survival was the most sensitive biological endpoint, with growth and reproduction somewhat less sensitive than survival. The no observed effect concentration (NOEC) for survival was 8mg/L, and the lowest observed effect concentration (LOEC) was 15mg/L dissolved arsenic. The LOEC for growth and reproduction was greater than the highest concentration tested, 56mg/L. Based on survival, the final chronic value (geometric mean of the NOEC and LOEC) was 11mg/L dissolved arsenic. The F(1) generation appeared to acclimate to the prior arsenic exposure of the parental generation and was significantly less sensitive than the parental generation. For example, survival for the F(1) generation through day 12 was 100% in 56mg/L dissolved arsenic, compared to 26% for the parental generation. Growth of the F(1) generation was significantly less than that of the parental generation across all concentrations including the control, indicating a generational difference in brine shrimp growth rather than an arsenic effect. This study represents one of the few full life cycle toxicity tests conducted with brine shrimp. PMID- 12550095 TI - A comparative analysis of the heavy metal loading of small mammals in different regions of Bulgaria I: monitoring points and bioaccumulation features. AB - Data on liver and body copper, zinc, lead, and cadmium content of small mammals (rodents and insectivorous) were collected and analyzed. Data comparisons were performed in two aspects: (1) points and years of monitoring; (2) monitor species bioaccumulations. Specific bioaccumulation features were observed in some of the monitor species. A method for comparative evaluation of heavy metal loads in the different species is proposed using data for liver and body contamination. The loads of Clethrionomys glareolus and Apodemus flavicollis were compared, and the data are in agreement with data from other authors in Central Europe. A correlation between heavy metal content in the food and liver of snow vole was established. The data demonstrate that two of the regions investigated in Rila Mountain National Park could be assumed to be background locations. Some possible reasons for the heavy metal contamination of the low-altitude region in Rila are discussed. Not very significant pollution was observed around industrial facilities. Correlations between heavy metal levels in zoomonitors and meteorological factors were established. PMID- 12550096 TI - A comparative analysis of the heavy metal loading of small mammals in different regions of Bulgaria II: chromosomal aberrations and blood pathology. AB - Heavy metal content was monitored in small mammals inhabiting mountain ecosystems and two industrial polluted regions in Bulgaria. Rodents (Microtus arvalis, M. rossiaemeridionalis, Clethrionomys glareolus, Pitymys subterraneus, Chionomys nivalis, Apodemus flavicollis, A. sylvaticus, and Mus macedonicus) were used as zoomonitors. Pathological changes in chromosome status, hematological indices, and blood cell morphology were analyzed in the context of heavy metal bioaccumulation. Significant correlations were obtained between the heavy metal load of zoomonitors and the frequency of chromosomal aberrations and pathological changes in erythrocytes (mainly micronuclei and basophilic granulations). It is suggested that mercury is a strong damaging factor for chromosomes and red blood cell apparatus. PMID- 12550097 TI - An iron-rich diet protects the liver and kidneys against cadmium-induced injury in the bank vole (Clethrionomys glareolus). AB - The objective of this study was to determine whether supplemental dietary iron (Fe) would protect against cadmium (Cd)-induced injury in the liver and kidneys of bank voles. The rodents were provided, for 6 weeks, Fe-adequate (60-80 microg/g) and Fe-enriched (250-270 microg/g) diets containing 0.05 (control), 40, and 80 microg Cd/g. Histological examinations and analyses of Cd, Cd bound and not bound to metallothionein (MT), Fe, and lipid peroxidation in liver and kidneys were carried out. The Fe-enriched diet prevented Cd-induced histopathological changes as well as deprivation of tissue Fe and lipid peroxidation. Also, supplemental Fe significantly decreased hepatic and renal Cd burden. However, in the Cd-80 bank voles fed the Fe-enriched diet, the non-MT bound Cd, considered a toxic species, reached 4.7 microg/g liver and 13.7 microg/g kidney, these values being similar to those at which histopathological changes occurred in the voles fed Cd diets not supplemented with Fe. The data indicate that the protective effect of supplemental Fe in the bank vole may be due to the prevention of Cd-induced deprivation of tissue Fe and Fe-dependent oxidative processes rather than to reduction of cadmium accumulation. PMID- 12550098 TI - Treated municipal sewage discharge affects multiple levels of biological organization in fish. AB - The objectives of this study were to examine cellular-, organ-, and organism level responses in longear sunfish (Lepomis megalotis) and fish community structure in a stream in which treated municipal sewage effluent is discharged and in a nearby reference stream with little surrounding land use. A modified version of the U.S.E.P.A. Rapid Bioassessment Protocol V, which combines a habitat assessment with Karr's index of biotic integrity, was used on 400-m reaches of each stream. The study site had a higher proportion of tolerant species and omnivores and a lower proportion of top predators, suggesting alterations in the fish community and a slight level of water quality impairment. Significant increases in condition factor, hepatosomatic index, serum testosterone, and plasma vitellogenin concentrations were observed in male sunfish collected from the study stream in comparison to fish collected from the reference stream. There were no differences between sites in hepatic expression of the 70-kDa stress protein (HSP70). In conclusion, effects were observed at cellular, organ, organism, and community levels of biological organization in fishes exposed to treated municipal sewage effluent. PMID- 12550099 TI - Exposure to 17 alpha-ethinylestradiol and bisphenol A--effects on larval moulting and mouthpart structure of Chironomus riparius. AB - The effects of the endocrine-disrupting chemicals 17alpha-ethinylestradiol (EE) and bisphenol A (BPA) on the development of the aquatic life-cycle stages (eggs to pupa) of Chironomus riparius were investigated. The test species was exposed to sublethal concentrations of EE and BPA (10 ng/L-1mg/L) and effects on larval weight and moulting success were recorded. In addition, three mouthpart structures (mentum, mandibles, and pecten epipharyngis) present on the head capsules of fourth-instar larvae were examined for structural deformities. Moulting was delayed and larval wet weight significantly reduced (P<0.05) at the highest treatment concentration (1mg/L) of both chemicals. No significant effect on either of these parameters was noted in the remaining treatments. However, deformities in the mouthparts of C. riparius were observed at very low exposure concentrations (10 ng/L), although the incidence of deformities was greater in the chironomids exposed to EE than BPA. Effects were mainly associated with the mentum, with statistically significant differences in median deformity score (Kruskal-Wallis P<0.001) recorded for both chemicals. At similar effect concentrations, an increased percentage of exposed animals had deformities of the pecten epipharyngis; however, little evidence of deformity was noted for the mandibles. At high concentrations, where moulting and wet weight were affected, no incidence of mouthpart deformity was noted. The relevance of these results in relation to the identification of an "endocrine-sensitive" endpoint for invertebrates is discussed. PMID- 12550100 TI - Effects of 17 alpha-ethinylestradiol on sexual development of the amphipod Hyalella azteca. AB - The effects of the synthetic estrogen 17alpha-ethinylestradiol (EE) on sexual development of the freshwater amphipod Hyalella azteca was investigated. Organisms were exposed in a multigeneration experiment to EE concentrations ranging from 0.1 to 10 microg/L and the development of both external and internal sexual characteristics were studied. Second-generation male H. azteca exposed from gametogenesis until adulthood to 0.1 and 0.32 microg EE/L developed significantly smaller second gnathopods. The sex ratio of the populations exposed to EE for more than two generations tended, although not statistically significantly, to be in favor of females. Histological aberrations of the reproductive tract, i.e., indications of hermaphroditism, disturbed maturation of the germ cells, and disturbed spermatogenesis, of post-F1-generation males were observed in all EE exposures. These findings provide evidence that sexual development of H. azteca is affected by exposure to sublethal concentrations of EE. PMID- 12550101 TI - Determination of heavy metals in crayfish by ICP-MS with a microwave-assisted digestion treatment. AB - Contamination of the American red crayfish, from the Guadiamar riverside, is due to the disastrous toxic spill that occurred on 25 April 1998 in the mining area of Aznalcollar-Los Frailes, Spain. A high concentration of heavy metals in the waters from the mine pool and their spill to the river Guadiamar was the cause of the destruction of a great number of animal and vegetable organisms. An ICP-MS method for the total determination of heavy metals (Cu, Zn, Pb, and Cd) in whole bodies of American red crayfish (Procambarus clarkii) was used. Metals were extracted from the matrix in a closed-vessel microwave digestion system with nitric acid and hydrogen peroxide. A study of the uncertainty of the method for the determination of metals was carried out; at a concentration of 5 microg/L, the uncertainty was below 34%. PMID- 12550102 TI - Growth responses of an estuarine fish exposed to mixed trace elements in sediments over a full life cycle. AB - Hatchling Cyprinodon variegatus were raised in the presence or absence of sediments contaminated with mixed trace elements to examine lethal and sublethal bioenergetic effects (metabolic rate, lipid storage, growth, reproduction) over a full life cycle (>1 year). Contaminated sediments were derived from a site receiving coal combustion residues (CCR) and were elevated in numerous trace elements including Al, As, Ba, Cd, Cu, Se, and V. Exposures were conducted at two levels of salinity (5 and 36 ppt) to examine the potential interaction of this variable with contaminants. Salinity had no effect on responses measured. Over the course of the study, fish exposed to contaminated sediment accumulated several CCR-related trace elements, including As, Cd, Se, and V. There were no differences in fish survival for contaminated sediment treatments and uncontaminated sediment treatments, nor were there differences in metabolic expenditures. However, growth, male condition factor, and storage lipid content in females were reduced due to contaminant exposure. No significant effects on fecundity or the proportion of females that were gravid at the end of the study were observed, yet females raised under control conditions produced 12% larger eggs than did females raised on contaminated sediments. During the presumably most-sensitive early life stages, individuals were not noticeably affected by contaminants, but rather the effects of exposure became apparent later in life. Because many species inhabit contaminated areas for long periods of time, often encompassing the entire life cycle, exposures focused only on specific life stages may substantially underestimate the overall responses elicited by individuals. PMID- 12550103 TI - Thrombin enhancement of interleukin-1 expression in mononuclear cells: involvement of proteinase-activated receptor-1. AB - In addition to its central role in blood coagulation and hemostasis, human alpha thrombin is considered a pro-inflammatory molecule. We have previously demonstrated that differentiated monocytes express the proteolytically activated receptor for thrombin (PAR-1) and that thrombin enhances the release of interleukin (IL)-6 in human monocytes. In the present study we show that thrombin upregulates the production of both IL-1alpha and IL-1beta in phytohemagglutin (PHA)-activated human peripheral blood mononuclear cells (PBMC). Treating PHA activated PBMC with the PAR-1 activation peptide, SFLLRN, mimics the effects of thrombin on IL-1alpha and IL-1beta production. Thus, it appears that these pro inflammatory effects induced by thrombin may be mediated through activation of PAR-1. ELISA and RNase protection assays indicate that thrombin and SFLLRN peptide upregulates IL-1 expression at both protein and mRNA levels. Thrombin directly affects monocyte IL-1 expression, since treatment of differentiated U937 cells with thrombin and SFLLRN enhances IL-1 production. These results may help explain how thrombin can enhance IL-1 expression in normal tissue to initiate tissue repair and why thrombin and thrombin-like enzymes may contribute to inflammatory responses observed in several pathophysiological conditions. PMID- 12550104 TI - IL-6 enhances IgE-dependent histamine release from human peripheral blood-derived cultured mast cells. AB - We examined whether interleukin (IL)-6 exerts the stimulatory effects on the secretion of histamine from human mast cells triggered by crosslinking of the high affinity IgE receptor (FcepsilonRI) with IgE and anti-IgE. As target cells, we used peripheral blood-derived cultured mast cells grown with SCF, because they were superior in FcepsilonRIalpha expression to cord blood-derived mast cells. Incubation with SCF+IL-6 for 1 week increased the IgE-dependent release as well as intracellular content of histamine in the cultured mast cells, as compared with the values obtained by incubation with SCF alone. The magnitude of these increases was higher than that for priming with SCF+IL-4. A striking difference was also found in the expression of FcepsilonRIalpha between the two-factor combinations. The addition of IL-6 during FcepsilonRI crosslinking with IgE/anti IgE in the presence of SCF did not influence histamine secretion. When SCF, IL-6 and IL-4 were used together, a further increase was observed in the anti-IgE dependent liberation of histamine from the cultured mast cells, compared with the two-factor combinations. These results suggest that IL-6 functions as a secretagogue for the inflammatory mediator of human mast cells in the presence of SCF. PMID- 12550105 TI - Peripheral mononuclear cell IL-10 and IL-12 production is not impaired in patients with advanced cancer and severe infection. AB - We assessed whether interleukin (IL)-10 and IL-12 production of blood mononuclear cells (PBMC) after lipopolysaccharide (LPS 0.1 microgram/ml, 1.0 microgram/ml) or phytohemagglutinin (PHA 5 microgram/ml) stimulation in advanced cancer patients with severe infections (IA, n=10) differs from those without infection (AC,n =10) or healthy controls HC (n=10). PBMCs in IA produced statistically higher median IL-12 levels than those in HC; LPS 0.1 (P=0.05), LPS 1.0 (P=0.019) and PHA (P=0.024). IL-10 production was always higher in AC than in HC. Both the IL-10 (LPS 1.0; P=0.018) and IL-12 (LPS 0.1;P =0.046) production of PBMCs were significantly higher in IA than one month afterwards. In conclusion, the PBMC production of IL-10 and IL-12 in advanced cancer patients with or without infection was not impaired but rather enhanced compared to that of HC. One month after infection, however, PBMC production clearly decreased, suggesting a kind of hyporeactivity. PMID- 12550106 TI - In vitro effects of eicosanoids derived from different 20-carbon Fatty acids on production of monocyte-derived cytokines in human whole blood cultures. AB - INTRODUCTION: Prostaglandins (PG) and leukotrienes (LT) are usually formed from arachidonic acid (e.g. PGE(2), LTB(4), LTC(4)). The anti-inflammatory effects of fish oil may be mediated through the production of alternative PG and/or LT formed from eicosapentaenoic acid (e.g. PGE(3), LTC(5)). This study examines the effects of PG and LT derived from different fatty acid precursors on lipopolysaccharide-induced cytokine production by cultured human whole blood. Methods Human whole blood was diluted 1:5 and incubated for 48h with lipopolysaccharide. PGE and LT were added and the concentrations of inflammatory cytokines in the cell culture supernatants determined. RESULTS: Tumour necrosis factor (TNF)-alpha concentrations were significantly decreased by the addition of PGE. At the maximum concentration used (10(-6)M) TNF-alpha concentration was reduced to 100%, 90% and 70% by PGE(1), PGE(2) and PGE(3) respectively. Likewise, interleukin (IL)-1beta concentration was decreased to 60%, 30% and 40% by 10(-6)M PGE(1), PGE(2) and PGE(3), respectively. IL-6 and IL-10 concentrations were not altered by PG. LTB(4), LTC(4) or LTC(5) did not significantly affect cytokine concentrations. CONCLUSIONS: PGE inhibit lipopolysaccharide-stimulated TNF-alpha and IL-1beta production in human whole blood cultures. PGE(1), PGE(2) and PGE(3) show a similar pattern and magnitude of effect. This suggests that the anti inflammatory effects of dietary fish oil may not be mediated through a simple substitution of one family of eicosanoids for another. PMID- 12550107 TI - Leukaemia inhibitory factor (lif) is expressed in hypertrophic chondrocytes and vascular sprouts during osteogenesis. AB - Avascular cartilage is replaced by highly vascularized bone tissue during endochondral ossification, a process involving capillary invasion of calcified hypertrophic cartilage in association with apoptosis of hypertrophic chondrocytes, degradation of cartilage matrix and deposition of bone matrix. All of these events are closely controlled, especially by cytokines and growth factors. Leukaemia inhibitory factor (LIF), a member of the gp130 cytokine family, is involved in osteoarticular tissue metabolism and might participate in osteogenesis. Immunohistochemical staining showed that LIF is expressed in hypertrophic chondrocytes and vascular sprouts of cartilage and bone during rat and human osteogenesis. LIF is also present in osteoblasts but not in osteoclasts. Observations in a rat endochondral ossification model were confirmed by studies of human cartilage biopsies from foetuses with osteogenesis imperfecta. LIF was never detected in adult articular chondrocytes and bone marrow mesenchymal cells. These results and other data in the literature suggest that LIF is involved in the delicate balance between the rate of formation of calcified cartilage and its vascularization for bone development. PMID- 12550108 TI - Different chemokines are expressed in human arthritic bone biopsies: IFN-gamma and IL-6 differently modulate IL-8, MCP-1 and rantes production by arthritic osteoblasts. AB - In the present study we analyse chemokine expression in the remodelling of subchondral bone in arthritis patients. Trabecular bone biopsies were tested by immunohistochemistry to identify interleukin (IL)-8, GRO-alpha, MCP-1, RANTES, MIP-1alpha and MIP-1beta expression. Subsequently, we evaluated by immunoassay the effect of interferon (IFN)-gamma and IL-6 on chemokine production by osteoarthritis (OA), rheumatoid arthritis (RA) and post-traumatic (PT) patients' isolated osteoblasts (OB). OB constitutively produced in situ IL-8, GRO-alpha, MCP-1, RANTES and MIP-1alpha. MIP-1beta was positive only in mononuclear cells. In RA many of these chemokines were also produced by mononuclear cells. IFN-gamma significantly down-regulated IL-8 and up-regulated MCP-1 produced by OB from all patients tested, whereas it did not affect the other chemokines analysed. Moreover, IFN-gamma reduced IL-1beta-stimulated IL-8 production but significantly increased both MCP-1 and RANTES. Interestingly, IL-6 significantly downregulated IFN-gamma-induced MCP-1 production, that was significantly lower in OA compared to RA patients. OB expressed chemokines both in vivo and in vitro suggesting that these cells are primary effectors in the bone capable of regulating autocrine/paracrine circuits that affect bone remodelling in these diseases. PMID- 12550109 TI - Transforming growth factor-Beta 1 (tgf-Beta 1) in patients with amyotrophic lateral sclerosis. AB - Previous investigations have shown that the transforming growth factor-beta 1 (TGF-beta 1) may protect neurons against excitotoxic and oxidative damage and may inhibit apoptosis. The aim of this study was to investigate the role of TGF-beta 1 in patients with amyotrophic lateral sclerosis (ALS). The study involved 24 ALS patients and 15 control group people. The ALS patients were divided into groups according to their clinical status, and duration of ALS. The TGF-beta 1 in the serum and cerebrospinal fluid (CSF) was measured by the enzyme-linked immunosorbent assay (ELISA). Results showed that TGF-beta 1 concentrations in the serum, and CSF in the whole group of ALS patients did not differ from those of the controls, but the serum TGF-beta 1 concentration was significantly higher in ALS patients with a terminal clinical status than in controls. The TGF-beta 1 concentration was significantly higher in the CSF of the patients, with a long duration of ALS, than in the patients with a short duration of ALS, and there was a significant positive correlation between the CSF TGF-beta 1 and the duration of ALS. TGF-beta 1 may play a role in neurodegeneration of ALS, and may be an indicator of the duration of the disease. PMID- 12550110 TI - TNFalpha and IL-1beta exert no direct vasoactivity in human isolated resistance arteries. AB - The proinflammatory cytokines TNFalpha and IL-1beta are thought to play a role in the mechanism of septic vasodilatory shock. Most evidence indicate that the vascular effects these cytokines are mediated through increased expression of inducible nitric oxide synthase, whereas several animal studies report a more rapid vasorelaxant effect. We studied the direct vascular effects of TNFalpha and IL-1beta in human isolated resistance arteries (n=9) using a microvascular myograph. After precontraction with noradrenaline or a depolarising high potassium solution, TNFalpha and IL-1beta (up to 10(-8) M, alone or in combination) did not exert any relevant vasoactive effect. We conclude that, in contrast with some animal experiments, these cytokines exert no direct vasoactivity in human resistance arteries. PMID- 12550111 TI - Vascularisation of tissue-engineered grafts: the regulation of angiogenesis in reconstructive surgery and in disease states. AB - Angiogenesis (the formation of new blood vessels) is essential for the growth of new tissue, tissue repair and wound healing. Tissue engineering, the construction of new tissue and organs for reparative purposes, relies on angiogenesis for the vascularisation of these new grafts. In tissue engineering, the emphasis to date has been on vascularisation of newly constructed tissue grafts by an extrinsic blood supply, and relatively little attention has been given to the possibility of building these grafts around an intrinsic blood supply. However, there are many disease processes, notably tumour growth, where excess angiogenesis can be a major problem. The purposes of this review are, first, to examine various methods of vascularising tissue-engineered grafts, and, second, to compare the role of angiogenesis in tissue engineering, where stimulation of angiogenesis is paramount, with pathological states, such as tumour growth, where angiogenesis needs to be inhibited. PMID- 12550112 TI - A follow-up study to investigate the efficacy of initial treatment of lentigo maligna with surgical excision. AB - Recurrence rates following conventional surgery for lentigo maligna (LM) are reported to be between 7% and 15%. However, the studies are few, contain small numbers of patients and have relatively short follow-up. The principle aim of this study was to determine the efficacy of conventional surgery for LM in a large unbiased sample of all LM presenting in a defined geographical area. All LM cases occurring in Leicestershire between 1987 and 1996 were identified. Data were gathered from case notes, general practitioners and the Office for National Statistics (for any mortality data) and patients were invited to attend for examination. There were 89 evaluable patients treated with primary excision, representing the largest reported series to date. There was a false positive rate for diagnostic biopsy for LM (when the correct diagnosis was lentigo maligna melanoma, LMM) of 5% (95% confidence interval, CI: 1% to 14%). The initial excision was histologically incomplete in 9% (4% to 17%) of cases. In completely excised lesions (n=81) the observed recurrence rate was 20% (CI: 12% to 30%) at a mean follow-up of 42 months, which is similar to previous reports. However, Kaplan-Meier analysis yielded an estimated probability of recurrence of 31% (CI: 19% to 50%); time to relapse was up to 66 months. The age and sex of the patient and the site of the lesion were not associated with outcome, but, surprisingly, smaller lesions were associated with incomplete excision and recurrence. Fifteen current lesions were excised with similar success rates: the incomplete re excision rate was 7% (CI: 0.2% to 32%) and the recurrence rate was 31% (CI: 12% to 83%) at 28 months. The estimated rate of transformation to LMM after initial surgical treatment was 1.5% (CI: 0.3% to 8%), and LMM was not the cause of death in any patient. These recurrence rates following complete primary excision and re excision following recurrence were high compared with general expectations, which is largely the result of the method of analysis and the long follow-up. The high rate calls into question the accuracy of the routine histological assessment of clearance. The development of LMM was rare following surgery. PMID- 12550113 TI - Incomplete excision of basal cell carcinoma: a prospective multicentre audit. AB - The audit of incomplete excision of basal cell carcinoma can be used as a parameter for clinical governance in plastic surgery units. However, there are no national standards, and all the previous reports from the UK have been retrospective and from regional units only. This 1 year prospective audit was undertaken simultaneously in the plastic surgery units of three different categories of hospital: a regional plastic surgery unit (University Hospital of South Manchester), a supraregional cancer hospital (Christie Hospital) and a district general hospital (Royal Bolton Hospital). A total of 757 lesions were excised from 600 patients, with 34 lesions (4.5%) being incompletely excised. The incidences of incomplete excision were similar in the regional unit (3.2%) and the district general hospital (3.1%), but higher in the supraregional cancer hospital (7.5%). The commonest site for incomplete excision was the eyebrow, followed by the postauricular area, the nose and the temple. There were no statistically significant differences in the distributions of the age and sex of the patients, the site and size of the lesions or the methods of repair between the three hospitals. However, there were significant differences in the distribution of syndromal, multiple and recurrent lesions, the grade of surgeon, and the clinical and histological subtypes. When the various confounding factors were adjusted by logistic regression, the variables most likely to have affected the incidence of incomplete excision were found to be grade of surgeon, minimal excision margin and histological subtype. PMID- 12550114 TI - The c-myc oncogene: use of a biological prognostic marker as a potential target for gene therapy in melanoma. AB - The c-myc oncogene has been shown to be overexpressed in a number of malignancies, and may play an important role in the pathogenesis of malignant melanoma. Previous prognostic studies have demonstrated c-myc overexpression in a range of cutaneous melanomas, and levels of c-myc oncoprotein expression have been shown to correlate with clinical outcome in both primary and secondary disease. The purpose of this study was to investigate the in vitro manipulation of c-myc expression using antisense oligonucleotides. The human melanoma cell lines A375M, Be11 and WM115 were treated with c-myc antisense oligonucleotides, and the cellular growth was compared with controls. Antisense oligonucleotides reduced the growth rate of all three cell lines, and produced a reduction in c myc gene expression as measured by flow cytometry. The growth inhibitions in the A375M, Be11 and WM115 cell lines at 72 h were 36.6%, 35.8% and 29.3%, respectively. Each of these was significantly different from control cultures (P<0.01). The c-myc antisense produced a mean 75% reduction in c-myc oncoprotein expression when compared with controls in the A375M cells (P<0.001), a 49% reduction in the Be11 cells (P<0.001) and a 28% reduction in the WM115 cells (P=0.005). This study demonstrates the importance of the c-myc oncogene in controlling melanoma growth. It suggests that blocking the expression of this gene, using an antisense approach, reduces melanoma cell growth, and may potentially provide a novel gene-therapy strategy for the treatment of advanced melanoma. PMID- 12550115 TI - The influence of timing on the functional and morphological result after nerve grafting: an experimental study in rabbits. AB - Clinical experience and experimental work in sheep have shown that a two-stage approach to restoring muscle function with a long nerve graft and free muscle grafting seems to be more beneficial than a one-stage approach. Based on a standardised experimental protocol, one-stage and two-stage nerve grafting approaches in rabbits were compared, and the actual differences in muscle force, together with morphological data, were calculated. In 20 rabbits the saphenous nerve was used as a 7 cm nerve graft. Animals were separated into two groups. In group 1, 10 rabbits underwent a one-stage approach to reinnervate the rectus femoris muscle. In the left hindlimb, the proximal end of the graft was coapted to the cut motor nerve branch of the vastus medialis muscle, and the distal end was coapted to the nerve branch of the rectus femoris muscle. In group 2, 10 rabbits underwent a two-stage approach, leaving the distal end of the nerve graft unconnected to the rectus femoris muscle in the first stage. In the second stage, this end was coapted to the freshly cut motor nerve branch of the rectus femoris muscle. After 15 months, the maximum tetanic tensions in the reinnervated rectus femoris muscle and the contralateral unoperated muscle were determined. The graft and the motor branch distal to the graft were biopsied in order to count the number of regenerated myelinated nerve fibres. The mean+/-s.d. maximum tetanic tensions in the reinnervated rectus femoris muscles were 10.6+/-4.9 N in group 1 and 21.4+/-1.1 N in group 2. Compared with the unoperated side, the muscle force following denervation and reinnervation was 38.3% in group 1 and 58.9% in group 2 (P=0.01). The mean+/-s.d. numbers of regenerated myelinated nerve fibres distal to the graft in the rectus femoris muscle branch were 737+/-340 in group 1 and 1487+/-1004 in group 2 (P=0.05). These results show that the neurotrophic effect of an immediately connected target organ is far outweighed by the adverse effect of the longer period of muscle denervation. Therefore, nerve grafting and muscle transplantation should not be performed in the same operation. PMID- 12550116 TI - Intraoperative evaluation of skin-flap viability using laser-induced fluorescence of indocyanine green. AB - Laser-induced fluorescence of indocyanine green (ICG) is a new method for evaluating skin perfusion, which is superior to conventional fluorescein angiography. In a prospective clinical study ICG fluorescence video-angiography was used for the intraoperative evaluation of skin-flap perfusion. The results of ICG imaging were compared with clinical outcome 1 week postoperatively. Intraoperative ICG filling defects were always associated with delayed wound healing. In 50% of the patients, the regions of sloughing and epitheliolysis corresponded accurately to the regions of dye-filling deficits. All of the flaps without ICG filling defects healed primarily. These results suggest that ICG fluorescence is a sensitive tool for assessing nutritive blood flow in pedicled skin flaps with and without an axial vessel. Future clinical studies are required to establish critical threshold fluorescence indices that correlate with skin viability in the postoperative course. PMID- 12550117 TI - Reconstruction of the auricle. AB - Between 1976 and 2002, 272 ear reconstructions were performed in our unit using the methods of Tanzer (n=38), Brent (n=156) and Nagata (n=78). We present this experience and our modifications of these methods, and describe our current practice. PMID- 12550118 TI - Long-term results of facial rejuvenation by carbon dioxide laser resurfacing using a quantitative method of assessment. AB - Carbon dioxide laser resurfacing has been widely reported as an effective method for treating facial rhytides. Few published series, however, have assessed the long-term results of this procedure. We prospectively reviewed our long-term results of perioral CO(2) laser resurfacing using an accurate, reproducible and quantitative method of evaluation. Wrinkle depth was measured using a silicone elastomer to provide an exact negative-replica mask of the perioral region. Simple light microscopy was then used to measure the depth of rhytides. This was carried out preoperatively, 6 weeks postoperatively and 2 years postoperatively, and the results were compared for 31 patients. CO(2) laser resurfacing achieved a highly significant mean reduction in wrinkle depth at 6 weeks of 91% (paired t test:P <0.00001), and this was well maintained at 2 years, when the mean reduction in wrinkle depth was 87% (paired t-test: P<0.00001). During the long follow-up, complications were few, with transient erythema being the most common; there were no cases of scarring. As in other series that have reported longer follow-up, we encountered three cases of minor hypopigmentation. We have used light microscopy on silicone moulds as an accurate method of assessing outcome after laser resurfacing of perioral rhytides. The early highly significant reduction in wrinkle depth was maintained at 2 years with minimal associated short-term or long-term morbidity. PMID- 12550119 TI - Effect of hypothermia on blood-flow responses in pedicled groin flaps in rats. AB - Cooling is widely used for preserving tissues such as kidneys before transplantation and for preserving extremities before replantation. Hypothermia has also been shown to be effective in the temporary storage of free flaps. However, in the intact living body, cooling can be damaging to tissue and the body system. We used a custom-designed clamping method (after flap elevation, occlusion and release of the flap-feeding artery) and continuous laser Doppler flowmetry to investigate the effects of hypothermia on blood flow and postocclusive reactive hyperaemia in the flaps. The animal model used was the partially elevated epigastric flap of adult Sprague-Dawley rats. In the hypothermia group (n=12), the core temperature and the flap temperature were allowed to fall during anaesthesia. At core temperatures of 34.58 degrees C and 338 degrees C and after rewarming of the rat, the feeding artery was occluded for 30 s and 120 s to observe the changes in blood flow and postocclusive reactive hyperaemia in the flap. In the control group (n=12), the core temperature was maintained at more than 378 degrees C throughout the experiment. To compare the flap blood-flow responses to occlusion of the femoral artery in the hypothermia group and the control group, the postocclusive reactive hyperaemia periods (i.e. blood flow above baseline after clamp release) were analysed. Statistical analysis of the responses showed that the magnitude (P=0.024), duration (P<0.001) and amplitude (i.e. peak flow) (P=0.037) of postocclusive reactive hyperaemia were significantly decreased in the hypothermia group. Our results suggest that hypothermia significantly decreases blood flow and postocclusive reactive hyperaemia in the rat epigastric flap. This may increase the risk of ischaemic flap complications unless rewarming is performed. PMID- 12550120 TI - Barium sulphate and soft-tissue radiology: allying the old and the new for the investigation of animal cutaneous microcirculation. AB - The study of microcirculation using angiography is essential to the advancement of flap and angiogenesis research in plastic surgery. Until the mid-1980s, barium sulphate was the most commonly used contrast material, although it did not provide optimal visualisation of the vascular tree. In 1986, a new technique using lead oxide was proposed, which permitted very high-quality imaging and rapidly became the technique of choice, despite its high toxicity. We reconsider the former technique of barium-sulphate injection and combine it with soft-tissue radiology using mammographic film to achieve a radiological definition similar to that obtained with lead oxide, and discuss the advantages and disadvantages of the two methods. We conclude that barium sulphate and the use of mammographic film is an accurate, simple and non-toxic method of analysing the cutaneous circulation in small animals. PMID- 12550121 TI - Closed-field titanium magnets for the retention of complex craniofacial prostheses. AB - Where reconstruction with autogenous tissue is contraindicated in craniofacial reconstruction, or declined by the patient, a prosthesis may be used. Originally made from latex, cellulose or acrylic resins, prostheses are now largely made of silicone. These prostheses were traditionally retained using adhesives, natural undercuts at retention sites or spectacles. However, these methods were unsatisfactory because of frequent loss of retention, skin reactions to adhesives or unnatural movements of the prosthesis. Osseointegration became a popular method of prosthesis retention, initially using a bar-and-clip device. The use of a bar attached to the implant has certain shortcomings, however, including rigidity of the prosthesis and the difficulty in maintaining hygiene at the implant site. We present our experience with the use of magnets to hold the prosthesis in place, which allows ease of use, improved hygiene and a more natural mobility of the prosthesis. PMID- 12550122 TI - Reconstruction of the natal cleft with a perforator-based flap. AB - Recurrent chronic conditions of the natal cleft, such as pilonidal sinuses, are difficult problems to treat. The deep natal cleft and the rolling effect of one buttock surface over the other contribute to the high recurrence rate. Wide excision of the affected area is the treatment of choice; to cover the defect many techniques have been described that flatten the natal cleft and shift the suture line away from the midline to try to reduce recurrence. These techniques include Z-plasties, rhomboid flaps, V-Y flaps, gluteus maximus myocutaneous flaps and others. Five patients with recurrent natal-cleft problems were operated on over a 1 year period. After wide excision of the natal cleft, the defect was reconstructed with a parasacral perforator-based flap. Preoperatively, a perforator situated superolaterally to the defect is identified with a Doppler probe; the flap is then designed horizontally around the perforator and, after elevation, is rotated 908 over the defect. The mean hospital stay was 5 days. Follow-up ranged from 3 months to 15 months, with no signs of recurrence. We believe this to be a simple and reliable technique, with the advantages of placing the scars away from the midline and flattening the natal cleft - factors that help to prevent recurrence. PMID- 12550123 TI - Ipsilateral free fibula transfer for reconstruction of a segmental femoral-shaft defect. AB - We describe an ipsilateral free fibula transfer for segmental femur reconstruction with venous interpositional grafts. Preparation and use of vessels at the recipient site for microvascular anastomosis is avoided. This represents a valuable modification of previously reported techniques, and should be considered as an alternative to conventional techniques, which regularly require more dissection at the reconstructive site. PMID- 12550124 TI - Groin dissections in epidermolysis bullosa: a report of groin dissection for the control of metastatic squamous carcinoma in patients with epidermolysis bullosa. AB - Epidermolysis bullosa predisposes to the development of squamous cell carcinoma. Despite regular surveillance, the majority of severely affected epidermolysis bullosa patients die from metastatic spread. There are no reports on regional control of metastatic spread. We describe our experience with two cases of groin dissection in epidermolysis bullosa. By anticipating the pitfalls in the perioperative care of patients with epidermolysis bullosa, we feel confident that they should make a full recovery. PMID- 12550125 TI - Streptococcal myositis: a lesson. AB - Streptococcal myositis is a rare, often fatal, acute infection of the muscle, caused by an invasive group A beta-haemolytic streptococcus. It is characterised by muscle necrosis without abscess formation, and, in contrast to necrotising fasciitis, does not primarily affect the subcutaneous tissue or skin. A young adult male presented with streptococcal myositis initially affecting the rectus femoris muscle of his left thigh. The symptoms, signs and management are discussed. Particular emphasis is given to the benefits of emergency CT scans to diagnose and delineate the extent of the disease. These scans may need to be repeated if the disease progresses. The four cornerstones of management are: early diagnosis using emergency CT scans; high-dose intravenous antibiotics; early aggressive surgical debridement; and intensive fluid and nutritional support. Published by Elsevier Science Ltd. All rights reserved. PMID- 12550126 TI - Local recurrence of breast cancer around a prosthesis dome. AB - We report a patient who, 2 years after mastectomy and breast reconstruction using a permanent expander, developed metastatic carcinoma around the filling port of the prosthesis. We believe this is the first description of such a condition, the differential diagnosis of which includes a silicone granuloma. PMID- 12550127 TI - Breast asymmetry as a result of congenital amniotic folds in the thoracoabdominal area. AB - Breast asymmetry is a relatively common condition. Mild breast asymmetry usually does not require correction. A rare case of breast asymmetry associated with congenital amniotic bands in the thoracoabdominal area was surgically corrected by transposition of the ectopic right breast and nipple-areola complex with Z plasty and thick split-skin grafting. The Ribeiro technique was used to treat the tuberous left breast. PMID- 12550128 TI - A rare case of a facial-nerve neurofibroma in the parotid gland. AB - The incidence of solitary neurofibroma of the facial nerve originating in the parotid region is extremely low. We report a case of a solitary neurofibroma in a 30-year-old male, who initially presented with a parotid mass without facial paresis or paralysis. A chain of small nodules had been palpable in the right parotid region for the previous 2-3 years. MRI and CT scans revealed several small ovoid lesions extending from the frontal margin of the parotid gland to the retromandibular region. The lesions were surgically removed. The main trunk of the facial nerve was adherent to the dorsal side of the largest nodule; however, this mass was resected atraumatically. Histopathological examination indicated neurofibroma. The incidence, presentation, diagnosis and surgical treatment of intraparotid neurofibroma are discussed and compared with those of Schwannoma. PMID- 12550129 TI - Anomalous anterior midline cervical tube. AB - Anterior midline cervical anomalies, excepting thyroglossal tract abnormalities, are rare. Two unusual and almost identical cases of an anomalous congenital tube connecting the chin to the upper sternum are described; only one previous similar report exists in the literature. In all three cases, the tubes occurred as isolated anomalies in otherwise healthy children. PMID- 12550130 TI - Inflammatory expansion of a breast augmented by a Trilucent implant. AB - We present a case in which a Trilucent soya-oil-filled breast prosthesis was removed because of rupture and subsequent massive breast enlargement. PMID- 12550131 TI - Spontaneous disappearance of refractory viral warts at distant sites following carbon-dioxide laser treatment. AB - We report the spontaneous disappearance of distant viral skin lesions following carbon-dioxide laser treatment of refractory viral warts. PMID- 12550132 TI - Watch out for the K-wire: painful experiences in two cases. AB - According to reviews in the literature, only a few case reports have mentioned the unusual migration of K-wires during orthopaedic surgery since 1991. This report emphasises the potential migration of the K-wire during plastic surgery in order to avoid the possible catastrophic complications. PMID- 12550133 TI - Videomicroscopy as a tool for counting hairs. PMID- 12550134 TI - The versatile cross-stitch. PMID- 12550135 TI - Polypropylene: a biocompatible alternative to metal foil for use as a prosthetic nail plate. PMID- 12550136 TI - Surgery by satellite: a new generation of self-harmers. PMID- 12550137 TI - Lasagne: a simple model to assess the practical skills of split-skin graft harvesting and meshing. PMID- 12550138 TI - Versatility of the semi-rigid ruler in plastic surgery. PMID- 12550139 TI - An effective self-retaining retractor for limb surgery. PMID- 12550140 TI - Capacitance coupling and diathermy burns. PMID- 12550141 TI - UK guidelines for the management of cutaneous melanoma. PMID- 12550142 TI - Management of aplasia cutis congenita of the scalp: a continuing enigma. PMID- 12550146 TI - Barriers to accessing health care: the perspective of elderly people within a village community. AB - Many initiatives which emphasise the consumerist stance of patients make the apparent assumption that patients have the knowledge and desire to exercise their consumer rights. This study explores the extent to which there is a consumerist ethos among elderly people in a village community, and the factors which influence the accessing of health care in the community. Using a mini ethnographic approach, nine key informants were observed and interviewed. Cultural factors were found to influence coping in health and illness, and in legitimising access to primary health care. No informant saw the need to exercise their rights as consumers of health care, suggesting that despite initiatives to involve patients as partners in health care, the hierarchical position of the elderly people in the village is unchanged from the days of the medical model in health care, and is a significant barrier to their use of health services. PMID- 12550145 TI - Acupressure and quality of sleep in patients with end-stage renal disease--a randomized controlled trial. AB - The purpose of the study is to test the effectiveness of acupressure on sleep quality of end-stage renal disease patients. The study was a randomized controlled trial; qualified patients in the dialysis centers of four major hospitals were randomly assigned into an acupressure group, a sham acupressure group, and a control group. A total of 98 participants were included in the study. The main outcomes measured were the Pittsburgh sleep quality index (PSQI) and the sleep log. Data were collected at pretreatment and following treatment. Primary statistical analysis was by means of Analysis of Covariance, the Kruskal Wallis Test and repeated measure ANOVA. The results indicated that PSQI scores of the acupressure group have a significantly greater improvement (p < 0.01) than the control group. However, there were no differences between the acupressure group and the sham group or the sham group and the control group (p > 0.05). Subscales of PSQI were further analyzed. Results demonstrated significant differences between the acupressure group and the control group in subjective sleep quality (p = 0.009), sleep duration (p = 0.004), habitual sleep efficiency (p = 0.001), and sleep sufficiency (p = 0.004). Significant differences in the subscale of subjective sleep quality (p = 0.003) between the sham acupressure group and the control group were also observed. Sleep log data showed that the acupressure group significantly decreased awake time and improved quality of sleep over time more than the control group (p < 0.01). The improvement could be seen as soon as the acupoints massage was implemented, and it was maintained through the post intervention. PMID- 12550147 TI - What factors influence Hong Kong school students in their choice of a career in nursing? AB - Chronic and recurrent shortages of nurses, coupled with the increasing demand for high quality and motivated nurses and competition for secondary school leavers from other professions, are issues which prompted this investigation of senior school students' career choice. Using a descriptive survey design, and a questionnaire developed for the study, 1246 Form 6 students in Hong Kong were surveyed, of these 28% respondents reported that they were interested in studying nursing. The findings indicated that students' decision to choose or not choose nursing was significantly influenced by the demographic factors: gender (chi(2) = 42.72, p < 0.0005), biology subject pursued (chi(2) = 18.94, p < 0.0005), previous academic achievement (chi(2) = 34.33, p < 0.0005) and mother's occupation (chi(2) = 21.08, p = 0.012); and their perception of nursing as a career score (t = 14.21, df = 1237, p < 0.0005). Students' intention to study nursing was also significantly affected by social influence: parents (chi(2) = 11.53, p = 0.001), school career masters (chi(2) = 5.52, p = 0.019) and friends (chi(2) = 4.83, p = 0.028); past experience with career activities (chi(2) = 84.479, p < 0.005); and working in hospital (chi(2) = 31.742, p < 0.005). Backward elimination logistic regression analysis concluded that being female (p < 0.0005), biology study at Form 5 and A-level (p < 0.0005), total perception scores (p < 0.0005), and participation in career activities (p < 0.0005) were found to be predictors of the students' intention to study nursing. Results of this study can be used by nurse leaders and recruiters to develop strategies and help school leavers perceive nursing in a more positive way. Expanding the informational sources about nursing can facilitate the recruitment process. PMID- 12550148 TI - Hospitalized children's descriptions of their experiences with postsurgical pain relieving methods. AB - The purpose of this study was to describe children's (aged 8-12 yr) experiences with postsurgical pain relieving methods, and their suggestions to nurses and parents concerning the implementation of pain relief measures in the hospital. The data were collected by interviewing children (N = 52) who were inpatients on a pediatric surgical ward in the university hospital of Finland. Content analysis was used to analyze the data. The children rated the intensity of pain on a visual analogue scale. The results indicated that all of the children used at least one self-initiated pain relieving method (e.g. distraction, resting/sleeping), in addition to receiving assistance in pain relief from nurses (e.g. giving pain killers, helping with daily activities) and parents (e.g. distraction, presence). The children also provided suggestions, especially as it relates to nurses (e.g. creating a more comfortable environment), regarding the implementation of effective surgical pain relief. However, some cognitive behavioral and physical methods were identified that should be implemented more frequently in clinical practice. Furthermore, most children reported their worst pain to be severe or moderate, which indicates that pain management in hospitalized children should be more aggressive. PMID- 12550149 TI - A comparison of effectiveness of bladder training and pelvic muscle exercise on female urinary incontinence. AB - This study compared the effectiveness of bladder training versus pelvic muscle exercises in the treatment of urinary incontinence in women. It was revealed that the two intervention groups showed improvement compared with the control group, but differed in their effects on outcome measures. The pelvic muscle exercise group was more effective in increasing the peak and the average pressures of pelvic muscle contraction. The bladder-training group was more effective in reducing urinary frequency and in increasing voided volume. Further research is needed to explore the relationship among the various outcome measures of urinary incontinence. PMID- 12550150 TI - Clinical teachers as caring mothers from the perspectives of Jordanian nursing students. AB - The purpose of this transcultural qualitative study was to discover, describe and explain the meaning of a caring student-teacher encounter within the context of clinical education. Clinical teachers are registered nurses who have completed a university undergraduate nursing degree program and at least 1 year of post registration clinical experience. They are employed as full-time staff of the faculty of nursing. Among the 19 Jordanian undergraduate nursing students interviewed, "clinical nurse teachers as caring mothers," emerged as an important theme. This paper describes the clinical teachers in their mothering roles, such as supporting, negotiating, reinforcing, transforming and releasing nursing students throughout their clinical practice. Understanding students' cultural beliefs and values provides possible predictors that could facilitate positive student-teacher relationships that could be used to plan the clinical education for nursing students. There is also a need to develop workshops in clinical teaching that would incorporate cultural awareness, especially in a multicultural student-teacher groups. PMID- 12550151 TI - Nurse-led intermediate care: patients' perceptions. AB - Intermediate care currently forms one of the UK Government's main initiatives for improving the quality of post-acute care. This paper examines patients' and carers' experiences of a nurse-led unit, which aims to provide intermediate care for people no longer acutely ill. Drawing on findings from qualitative interview data, we demonstrate that patients viewed this model of care as acceptable but that they had markedly inconsistent experiences of care and reported considerable variation in their perceptions of the Unit's purpose. Some possible reasons for this are explored. Implications for the development of good quality nurse-led intermediate care are outlined. PMID- 12550152 TI - The Yorkshire BARRIERS project: diagnostic analysis of barriers to research utilisation. AB - The study identified barriers to research implementation experienced by nurses, midwives and health visitors in five trusts and one health authority in Yorkshire, UK. Funk et al. (Appl. Nurs. Res. 4(1) (1991a) 39, Appl. Nurs. Res. 4(2) (1999b) 90) developed the BARRIERS to research utilisation questionnaire over 10 years ago, but no replication, in size (n = 1989) and extent, of that study appeared to have occurred. The staff population (n = 4501) were sent the BARRIERS questionnaire. 44.6% (n = 2009) were returned. Findings suggested nurses need time to read and apply research; authority to change practice; critical appraisal skills, an understanding of statistics and support of managers and peers (particularly doctors) to achieve successful practice change. PMID- 12550153 TI - Health needs assessment in the health visiting service and the impact on the ethnic community. AB - The purpose of this article is to share the experience of using a health needs assessment tool in health visiting practice in Great Britain, with clients who do not speak English. This is an important issue in developing equity of practice across the growing multi-cultural and diverse populations of the United Kingdom. The paper outlines the findings relevant to these issues drawn from a wider study that used qualitative methods to observe and interview both health visitors and clients regarding the use of the tool. It focuses on one vignette to demonstrate the practical and difficult issues when a formal system is used to assess needs in clients who do not speak English as a first language. PMID- 12550735 TI - Treatment options for status epilepticus. AB - Status epilepticus is a neurological emergency requiring prompt pharmacological intervention. Recent advances in the treatment of this condition include the introduction of treatment algorithms that are tailored more specifically to clinical situations, a trend towards more aggressive therapies if initial treatment with front-line agents fail, and a better understanding of the role of treatment for patients in status epilepticus in the out-of-hospital setting. PMID- 12550736 TI - GABA(A) receptor function and pharmacology in epilepsy and status epilepticus. AB - GABA(A) (gamma-n-aminobutyric acid) receptor dysfunction has long been implicated in the development of epilepsy and status epilepticus. Recent advances have been made in understanding the cellular, pharmacological and genetic involvement of GABA(A) receptors in seizure disorders. In particular, genetic mutations found in GABA(A) receptor subunits strongly implicate the GABA(A) receptor in idiopathic generalised epilepsies. PMID- 12550737 TI - Recent developments from genetic mouse models of seizures. AB - The use of genetically altered mice has revolutionised biomedical research into the genetics and neurobiological mechanisms contributing to complex human disorders such as epilepsy. Recent studies using mutant mice have expanded our knowledge of the key roles that abnormalities in synaptic function and formation play in epileptogenesis, and further illustrate just how closely some mouse models resemble human epilepsy. Broader utilisation of epileptic mouse mutants should provide new molecular targets for developing novel anti-epileptic drugs, and also improved means for predicting their efficacy in currently refractory forms of epilepsy. PMID- 12550738 TI - Pharmacology of traumatic brain injury. AB - The intensity of experimental and clinical research to identify a neuroprotective drug for the treatment of traumatic brain injury is motivated by the devastating morbidity and mortality of this condition. Encouraging experimental work has led so far to disappointing clinical trials and the identification of new potential therapeutic targets is critically dependent on a better understanding of the chronic pathophysiology triggered by the initial insult. Future advances in the pharmacological treatment of traumatic brain injury are likely to include the evaluation of sequentially timed therapies combining multiple and targeted agents, and manipulation of the newly discovered neurogenic potential of the adult brain together with the refinement of traditional interventions to block specific cytotoxic cascades. PMID- 12550739 TI - Brain plasticity and pathology in psychiatric disease: sites of action for potential therapy. AB - Recent advances have been made in understanding the changes in neuronal plasticity in psychiatric disease at the molecular level (changes in neurotransmission, long-term potentiation, long-term depression, glutamate receptors, synaptic strength and neurotrophic support) and the systems level (changes in hippocampal, frontal and amygdala function in health and disease and the impact of stress on the hippocampal/frontal axis), as well as in the impact of drugs and sites of action for therapeutic intervention. Drugs acting on plasticity could affect the abnormal set point in psychiatric disease, resulting in treatment of the disease rather than just the symptoms. PMID- 12550740 TI - GDNF and somatostatin in sensory neurones. AB - Somatostatin (SOM) is a regulatory peptide produced throughout the central nervous system and in most major peripheral organs. In humans, the SOM stable analogue octreotide (OCT) is used to treat opioid-resistant pain. In animal models, SOM and OCT are analgesics and SOM released from the peripheral endings of sensory neurones exerts anti-inflammatory actions. The expression of SOM in sensory neurones can be modulated by the trophic factor glial-cell-line-derived neurotrophic factor (GDNF). Recent data show that GDNF modulates activity-induced release of endogenous SOM from both central and peripheral terminals of adult sensory neurones. This novel mechanism deserves exploration as a potential new therapeutic strategy to control two major features of inflammation: pain and leukocyte recruitment. PMID- 12550741 TI - Functional interactions between cannabinoid and metabotropic glutamate receptors in the central nervous system. AB - The recent appreciation that two G-protein-coupled receptors, metabotropic glutamate and cannabinoid, are trans-synaptically linked by a small lipid messenger has profound implications, both for control of synaptic transmission and for novel therapeutic strategies. There is much evidence for this assertion and on the significance of this dual receptor cooperation for modulation of synaptic transmission in the central nervous system. PMID- 12550742 TI - CB1 cannabinoid receptor signalling in Parkinson's disease. AB - Signalling at CB(1) cannabinoid receptors plays a key role in the control of movement in health and disease. In recent years, an increased understanding of the physiological role of transmission at CB(1) receptors throughout the basal ganglia circuitry has led to the identification of novel therapeutic approaches to both the symptoms of Parkinson's disease and the side effects of current anti parkinsonian therapies, especially L(3,4) dihydroxyphenylalamine (levodopa) induced dyskinesia. Thus, because activation of basal ganglia CB(1) receptors can modulate neurotransmission and contribute to synaptic plasticity in a manner similar to that described in other brain regions, it also appears that endocannabinoids might modulate cell-cell signalling via effects on neurotransmitter re-uptake and postsynaptic actions mediating cross talk between multiple receptor types. Recent studies in animal models and in the clinic suggest that CB(1) receptor antagonists could prove useful in the treatment of parkinsonian symptoms and levodopa-induced dyskinesia, whereas CB(1) receptor agonists could have value in reducing levodopa-induced dyskinesia. PMID- 12550743 TI - CB2 cannabinoid receptor agonists: pain relief without psychoactive effects? AB - Cannabinoid receptor agonists significantly diminish pain responses in animal models; however, they exhibit only modest analgesic effects in humans. The relative lack of efficacy in man may be because of the dose limitations imposed by psychoactive side effects. Cannabinoid agonists that selectively target CB(2) (peripheral) cannabinoid receptors should be free of psychoactive effects, perhaps allowing for more effective dosing. CB(2) receptor activation inhibits acute, inflammatory and neuropathic pain responses in animal models. In preclinical studies, CB(2) receptor agonists do not produce central nervous system effects. Therefore, they show promise for the treatment of acute and chronic pain without psychoactive effects. PMID- 12550744 TI - Gene therapy in psychiatric disorders: too early, too complex? AB - Gene transfer strategies are being tested in a variety of animal models for psychiatric disorders. These promise to translate viral-mediated, non-viral and combinatorial techniques for delivery of transgenes into neuroanatomical and cell type-specific therapeutic tools. However, these disorders involve complex functional neurocircuits and developmental aetiologies that may present even greater challenges than other neurological conditions. PMID- 12550745 TI - The neuronal versus vascular hypothesis of migraine and cortical spreading depression. AB - Our understanding of the pathophysiological mechanisms of migraine remains poor despite the availability of clinically effective drugs and many years of research. Historically, two independent theories regarding the aetiology of headache were suggested: vascular and neuronal. However, recent data demonstrate that neuronal excitation modulates both the pial and meningeal circulation through a critical interaction with the trigeminal nerve, supporting the concept that the integration of neuronal and vascular information in the trigeminovascular network represents a key event in the aetiology of migraine. PMID- 12550746 TI - Cytokines: regulation of the hypothalamo-pituitary-adrenocortical axis. AB - Many of the pro-inflammatory cytokines that are released in response to immune/inflammatory insults exert marked stimulatory influences on the hypothalamo-pituitary-adrenocortical axis. Thus, they provoke the release of glucocorticoids that, in turn, temper the ensuing immune/inflammatory response, and thereby complete a homeostatic neuroendocrine loop. The mechanisms by which cytokines cause glucocorticoid release are complex and can be affected by repeated or sustained cytokine exposure, gender and age, or counter-regulatory mechanisms. PMID- 12550747 TI - The MCH receptor family: feeding brain disorders? AB - The importance of melanin concentrating hormone (MCH) in the control of energy balance has been confirmed by findings of lean phenotypes of mice with targeted deletion of the melanin concentrating hormone receptor 1 (MCH1-R). The recent publications of anorectic and antiobesity effects of the first two selective MCH1 R antagonists have confirmed the notion that pharmacological blockade of MCH1-R is a viable therapeutic approach for obesity. In addition, MCH1-R antagonists have been found to have antidepressant and anxiolytic properties. PMID- 12550748 TI - Trace amine receptors as targets for novel therapeutics: legend, myth and fact. AB - Trace amines are attracting attention as neurotransmitters because they are believed to play a role in human disorders such as schizophrenia, depression, attention deficit disorder and Parkinson's disease. Research to date is promising and confirms the need for continuing work to forge the way for new drug discovery. PMID- 12550749 TI - Active drug metabolites in drug development. AB - Active drug metabolites discovered during the course of drug development constitute a subset of the larger issue of metabolic drug interactions, but still demand unique consideration from both an efficacy and a safety point of view. Improved technology has allowed better identification and quantification of metabolites, raising new issues to be addressed during the course of drug development. Several new molecular identities recently entering the marketplace have active metabolites, and many more are (or have been) in development. The MIST (Metabolites in Safety Testing) committee of PhRMA (Pharmaceutical Research and Manufacturers of America) has prepared a position paper (in press) on the subject, which has been widely discussed (with and by regulatory authorities) over the past three years. PMID- 12550750 TI - Excitotoxicity, apoptosis and neuropsychiatric disorders. AB - In recent decades there has been an upsurge of interest in mechanisms of cell death in the central nervous system. Much attention has been focused on two cell death processes - excitotoxicity and apoptosis - and much confusion has arisen regarding how one relates to the other, and how either relates to neurological disorders. Recent findings shed new light on this issue, especially with respect to in vivo apoptosis, and views that are contrary to prevailing interpretations are expressed, especially in the burgeoning apoptosis field of inquiry. PMID- 12550751 TI - Updates on the cytogenetics and molecular genetics of bone and soft tissue tumors. Dermatofibrosarcoma protuberans and giant cell fibroblastoma. PMID- 12550752 TI - Different mechanisms lead to a karyotypically identical t(20;21) in myelodysplastic syndrome and in acute myelocytic leukemia. AB - A new t(20;21)(q11;q11), associated with a deletion on the long arm of chromosome 20, was found in one patient with an acute myelocytic leukemia (AML) and in one with myelodysplastic syndrome (MDS). In both cases deletion was interstitial, extending from band q11 to band q13, as shown by comparative genomic hybridization (CGH) and fluorescence in situ hybridization (FISH). FISH analysis with whole arm paints, subtelomeric probes, and locus-specific probes for the long arms of chromosomes 20 and 21 revealed in patient 1 a reciprocal translocation between the deleted 20q and the long arm of chromosome 21, that is, del(20)(q11q13)t(20;21)(q11;q11), and in patient 2, material from 21q was inserted into the deleted 20q, that is, del(20)(q11q13)ins(20;21)(q11;q11q22). This is the first identification of a complex 20;21 rearrangement in MDS/AML. Deletion at 20q and juxtaposition between 20q11 and 21q11 appear to be the critical genomic events. PMID- 12550753 TI - Chromosome 6 abnormalities are recurrent in synovial chondromatosis. AB - Synovial chondromatosis, a lesion composed of multiple nodules of cartilage involving articular or tendon sheath synovial membranes, has traditionally been considered a metaplastic condition. A specific or characteristic chromosomal anomaly has not yet been identified in synovial chondromatosis. Cytogenetic and molecular cytogenetic analyses of three cases of synovial chondromatosis revealed clonal karyotypic abnormalities in all three cases including structural abnormalities of chromosome 6 in two. Anomalies of chromosome 6 have been observed in three of five previously reported synovial chondromatosis cases. These findings support a neoplastic origin for synovial chondromatosis and suggest that chromosome 6 aberrations are recurrent in this lesion. PMID- 12550754 TI - Polymorphisms and mutations found in the regions flanking exons 5 to 8 of the TP53 gene in a population at high risk for esophageal cancer in South Africa. AB - A previous study in esophageal cancer (EC) patients from South Africa showed that 17% of tumors contained somatic mutations, including small deletions, insertions, and point mutations, resulting in frameshifts or amino acid changes in exons 5-8 of the TP53 gene. In the present study, polymerase chain reaction single-strand conformation polymorphism and DNA sequence analysis were used to search for sequence variation in the regions flanking exons 5-8 in a series of 74 primary human esophageal squamous cell carcinomas (ESCC). DNA from blood from 37 of the same EC patients and 118 blood samples from the same ethnic group, originating from the Transkei region of South Africa, a high-risk area for EC, served as controls. Mutations were rarely found in the regions flanking exons 5-8, but polymorphisms were frequent. Two mutations (G-->A, codon 331; G-->T, donor splice site) were found in the exon 9 region, while four polymorphisms occurred in intron 3 (16 bp duplication) and exon 4 (C-->A, codon 34; G-->C, codon 36; G-->C, codon 72) regions. Loss of heterozygosity occurred for the 16 bp polymorphism in the EC patients, but not in the controls. Certain genotypes were common in the EC group while others were common in the control group. Graphic representation illustrates the various mutations/polymorphisms found in the TP53 gene in samples from EC patients from South Africa. The results indicate that various small deletions and insertions occur at direct repeat sequences and can be explained by slipped mispairing. The point mutations include the polymorphism in codon 72 (Arg ->Pro), which has recently been associated with an increased risk of developing human papilloma virus-associated cancers. PMID- 12550755 TI - Interphase fluorescence in situ hybridization analysis of del(11)(q23) and del(17)(p13) in chronic lymphocytic leukemia. a study of 40 early-onset patients. AB - Although B-cell chronic lymphocytic leukemia (B-CLL) is the most common form of leukemia in Western countries, little is known about its underlying molecular abnormalities and their prognostic significance, particularly for use in early therapeutic interventions in young patients. As TP53 tumor suppressor gene abnormalities and 11q23 deletions are reported to be prognostically adverse in hematologic malignancies, we used interphase fluorescence in situ hybridization to analyze their incidence and prognostic significance in young B-CLL patients. Bone marrow samples from 40 untreated B-CLL patients at diagnosis were studied using five yeast artificial chromosome clones from the 11q23.1 approximately q23.3 chromosomal region and a probe specific for the 17p13.1 locus. Twenty-three patients (58%) carried 11q deletions. Interestingly, 16 of 17 patients (94%) who showed early disease progression exhibited this chromosomal abnormality, suggesting that 11q deletions may help to identify more aggressive disease in early stage patients. In contrast, monoallelic TP53 deletions were found in all of the patients. The TP53 and 11q deletions were only present in a proportion of the clonal B-cells, which suggests that they are secondary events in B-CLL. PMID- 12550756 TI - Genotypic analysis of primary and metastatic cutaneous melanoma. AB - Microdissection genotyping was performed on 16 cases of melanoma, including two cutaneous and one lymph node metastases. Three benign nevi were used as controls. Where possible, tumor was microdissected at several sites. Genotyping involved assessment of loss of heterozygosity [LOH]), which was accomplished using a panel of nine polymorphic tetranucleotide microsatellites. Polymerase chain reaction was performed on the normal tissue sample to establish microsatellite heterozygous status. Informative markers were then tested on microdissected lesional tissue and scored for the presence and extent of allelic imbalance (AI). Microsatellite informativeness varied from 33% to 66%. Benign nevi were without AI. All invasive melanomas manifested acquired allelic loss, which involved 75% or 100% of the markers shown to be informative for each subject. Eleven of 13 (84%) primary melanomas demonstrated intratumoral heterogeneity of AI consistent with development of tumor subclones with differing genotypic profiles within thin as well as thick melanomas. Although a consistent pattern did not emerge among the markers, LOH of 9p21 (D9S254) occurred in 60% (9/15) of the cases followed by 40% of cases displaying LOH of 1p34, p53, 10q (MXI1), and 10q23 (D10S520) and 25% with 5q21 (D5S 592) abnormalities. A third of the cases including the metastatic foci demonstrated two different patterns of AI affecting alternative alleles of the same genomic marker within different parts of the melanoma. Two melanomas in situ did not display LOH of any markers in the informative cases although the in situ component in the invasive tumors had allelic losses that were in part similar to the invasive areas. The results of this study support the expanded use of microdissection genotyping and explore other markers to define the unique mutational profile for malignant melanoma that may complement other histologic characteristics of melanoma. PMID- 12550757 TI - Evidence for another tumor suppressor gene at 17p13.3 distal to TP53 in hepatocellular carcinoma. AB - Loss of 17p is one of the most frequent chromosomal alterations in primary hepatocellular carcinoma (HCC). In the present study, the association between loss of 17p and TP53 mutation was analyzed in 94 primary HCC of Chinese patients. Loss of one allele at 17p13.3 distal to the TP53 gene was observed in 48 of 94 HCC (51%), whereas loss of heterozygosity (LOH) at 17p13.1 near the TP53 gene was detected in 30 of 94 HCC (32%) and TP53 mutation was detected in only 22 of 94 HCC (23%). High frequency of LOH at 17p13.3 and relatively low frequency of TP53 mutation in the present study indicate that loss of function of a putative tumor suppressor gene at 17p13.3 may play a more important role than TP53 in HCC development. PMID- 12550758 TI - Detection of t(11;18)(q21;q21) in marginal zone lymphoma of mucosa-associated lymphocytic tissue type on paraffin-embedded tissue sections by using fluorescence in situ hybridization. AB - A chromosomal t(11;18)(q21;q21) recurrently found in marginal zone lymphomas of mucosa-associated lymphocytic tissue (MALT) type is one of the key determinants of their treatment and outcome because t(11;18)-positive cases are resistant to Helicobacter pylori eradication therapy. To detect t(11;18) on paraffin-embedded tissue sections, we established a method of dual-color fluorescence in situ hybridization (D-FISH) using DNA probes directly labeled with SpectrumGreen and SpectrumOrange (tissue-FISH [T-FISH]). T-FISH detected the t(11;18) as colocalized signals between the band-specific yeast artificial chromosome (YAC) clones y966e4 (11q21) and y943b8 (18q21) on the der(11)t(11;18). The t(11;18) was detected in four of 22 patients with marginal zone lymphoma of MALT type in 72% 78% of the cells. In both patients studied, the percentage of t(11;18)-positive cells was much higher in tissue-FISH analysis than in interphase-FISH: 74% versus 30% in patient 1 and 75% versus 31% in patient 4. None of the 12 patients with diffuse large B-cell lymphoma of the stomach showed the t(11;18). Tissue-FISH is a powerful tool for the diagnosis of marginal zone lymphoma of MALT type because it can be applied not only to small specimens obtained from endoscopic biopsy samples but also to archival materials, hence facilitating the delineation of subtypes in marginal zone lymphoma of MALT type. PMID- 12550759 TI - Detection of a novel reciprocal t(16;22)(q11.2;q12) in a Ewing sarcoma. AB - Several structural and numerical chromosomal abnormalities have been identified as primary and secondary chromosomal aberrations in Ewing sarcoma (ES). The majority of these are t(11;22) and trisomies, especially of chromosome 8. Specific chromosomal abnormalities often correlate with particular morphologic or phenotypic subtypes of tumor and play an important role in prognosis. The objective of this report is the cytogenetic evaluation of a case of ES using G banding, fluorescence in situ hybridization, and spectral karyotyping techniques. Multiple chromosomal aberrations were identified including a novel reciprocal t(16;22)(q11.2;q12). PMID- 12550760 TI - AML1 amplification in a child with acute lymphoblastic leukemia. AB - We report a case of de novo acute lymphoblastic leukemia with tandem amplification of the AML1 gene located in a chromosome marker that originated from chromosome 21 and a long event-free survival. PMID- 12550761 TI - Spindle-cell rhabdomyosarcoma with 2q36 approximately q37 involvement. AB - The cytogenetic analysis of a spindle-cell variant of embryonal rhabdomyosarcoma (RMS), presenting as a cheek mass in an 18-year-old girl, is reported. The tumor cells showed an abnormal karyotype 46,XX,der(2)t(2;7)(q36 approximately q37;q3?),del(14)(q24),der(16)t(1;16)(q21;q13), with a tetraploid range of chromosome number in a subpopulation of cells. By fluorescence in situ hybridization analysis, the tumor cells were negative for FKHR-disrupting translocations specific for alveolar type of RMS and for NMYC gene amplification. PMID- 12550762 TI - Trisomy 8 as the sole chromosomal aberration in myelocytic malignancies: a multicolor and locus-specific fluorescence in situ hybridization study. AB - Trisomy 8 is the most common chromosomal aberration in myelocytic malignancies, occurring both as a sole change as well as in addition to other abnormalities. In spite of this, next to nothing is known about its pathogenetic importance or its molecular genetic consequences. Possible mechanisms involved in the transformation process include dosage effects of genes mapping to chromosome 8 and presence of specific mutations or cryptic fusion genes on the duplicated chromosome. In the latter case, +8 would be secondary to a cryptic primary rearrangement and not involved in leukemogenesis as such, but rather in tumor evolution. Although hidden genetic changes have been found in some trisomies, for example, mutations in KIT in acute myelocytic leukemia (AML) with +4 and in MET in hereditary papillary kidney carcinoma with trisomy 7, none associated with +8 have so far been discovered. To address this issue, we have investigated a total of 13 cases of AML, myelodysplastic syndromes, and chronic myeloproliferative disorders with trisomy 8 as the sole chromosomal anomaly. All cases were studied by combined binary ratio multicolor fluorescence in situ hybridization (FISH) and with FISH using locus-specific probes for both arms of chromosome 8, the subtelomeric regions of 8p and 8q, and the leukemia-associated genes FGFR1, MOZ, ETO, and MYC. No cryptic changes were detected, thus excluding the possibility of gross genetic rearrangements or aberrations involving these loci on chromosome 8. PMID- 12550763 TI - Genetic susceptibility to oral cancer and the expression of common fragile sites. a study of 100 patients. AB - The expression of bleomycin-induced fragile sites (FS) in the blood lymphocytes of 150 individuals (100 oral cancer patients and 50 age and sex matched controls) is described. FS expression frequencies in oral cancer patients were significantly higher when compared with controls. FS expression was site specific in oral cancer patients. Chromosome 5 was the most affected, with four of its FS expressing in high frequencies. Enhanced expression of FS at the centromeric region was observed in the patient group. This study emphasizes the role of FS in the genetic susceptibility to oral cancer. PMID- 12550764 TI - Chromosomal imbalances in pleomorphic rhabdomyosarcomas and identification of the alveolar rhabdomyosarcoma-associated PAX3-FOXO1A fusion gene in one case. AB - Rhabdomyosarcomas (RMS) are soft tissue sarcomas resembling developing skeletal muscle, and pleomorphic rhabdomyosarcomas (PRMS) are a rare nonpediatric entity. Little molecular cytogenetic information exists for PRMS, and their relationship to other subtypes of rhabdomyosarcoma and other sarcomas is unclear. Chromosomal imbalances were determined in seven well-characterized cases of PRMS using comparative genomic hybridization. The smallest overlapping regions of gain were 1p22 approximately p33 (71%), 7p (43%), 18/18q (43%), and 20/20p (43%), and the regions of loss were 10q23 (71%), 15q21 approximately q22 (57%), 3p, 5q32 approximately qter, and 13 (all 43%). Four of the seven cases had amplicons involving the regions 1p21 approximately p31, 1q21 approximately q25, 3p12, 3q26 approximately qtel, 4q28 approximately q31, 8q21 approximately q23/8q, and 22q. These regions are distinct from those frequently associated with the alveolar subtype, whereas the embryonal subtype without anaplasia is rarely associated with amplification events other than gain/amplification of 8q material. The regions of imbalance appeared more similar to those reported for malignant fibrous histiocytomas (MFH) and osteosarcomas, consistent with the suggestion that PRMS can be considered part of the spectrum of MFH. In addition, one of the cases classified as PRMS showed evidence for the presence of a PAX3-FOXO1A fusion gene, which is characteristic of the alveolar subtype of RMS. PMID- 12550765 TI - Derivative (7)t(7;8)(q34;q21). a new additional cytogenetic abnormality in acute promyelocytic leukemia. AB - Cytogenetic abnormalities in acute myelocytic leukemia (AML) have been identified as one of the most important prognostic factors. The t(15;17) is associated with high rates of complete remission and event-free survival. Secondary chromosomal changes are also present in approximately one third of patients with newly diagnosed acute promyelocytic leukemia (APL). Indeed, the gain of whole chromosome 8 may be involved in the course of APL under C-MYC gene dosage effect theory. Complete or partial loss of the long arm of chromosome 7 region has been recognized in preleukemic myelodysplasia or unfavorable AML. We report here two original APL cases in which a new additional chromosomal abnormality, der(7)t(7;8)(q34;q21), is associated with the t(15;17)(q22;q21). This recurrent abnormality results in a partial loss of 7q associated with a partial 8q trisomy. As the 7q and 8q breakpoints were similar in both cases, the involvement of these critical regions in the pathogenesis and outcome of APL disease has to be determined. PMID- 12550766 TI - Cytogenetic findings and clinical course in a consecutive series of Wilms tumors. AB - Wilms tumor (WT) is characterized by a nonrandom pattern of chromosome aberrations, but the clinical significance of different cytogenetic patterns is unknown. The present study describes the cytogenetic findings and the clinical course in a cohort of 39 children with WT. Samples for short-term culturing and cytogenetic analysis were obtained during a 15-year period. Clonal chromosome aberrations were detected in 23 samples from 19 patients. Tumors that relapsed more often showed clonal aberrations than did tumors that did not. However, this association my have been due to sampling bias. Among the cases with karyotypically abnormal samples, the modal chromosome number was in the near diploid range in 10, hyperdiploid/hypotriploid in 8, and hypodiploid in 1. The most common changes were trisomy 12 and gain of 1q material (8 cases each), trisomy/tetrasomy 8 (7 cases), and trisomy 13 (5 cases). None of these frequently occurring abnormalities, or the ploidy level, showed any association with clinical outcome, using tumor relapse as an end-point. Nor could any relationship between cytogenetic features and histopathologic subtype be discerned. Although the number of informative cases was too small for proper evaluation, the present study did not contradict the previous notion that loss of material from the long arm of chromosome 16 is associated with poor clinical outcome. All three patients with deletion of 16q developed metastases. PMID- 12550767 TI - Amplification of 17p11.2 approximately p12, including PMP22, TOP3A, and MAPK7, in high-grade osteosarcoma. AB - Amplification of region 17p11.2 approximately p12 has been found in 13%-29% of high-grade osteosarcomas, suggesting the presence of an oncogene or oncogenes that may contribute to their development. To determine the location of these putative oncogenes, we established 17p11.2 approximately p12 amplification profiles by semiquantitative PCR, using 15 microsatellite markers and seven candidate genes in 19 high-grade osteosarcomas. Most of the tumors displayed complex amplification profiles, with frequent involvement of marker D17S2041 in 17p12 and TOP3A in 17p11.2 and, in some cases, very high-level amplification of PMP22 and MAPK7 in 17p11.2. Our findings suggest that multiple amplification targets, including PMP22, TOP3A, and MAPK7 or genes close to these candidate oncogenes, may be present in 17p11.2 approximately p12 and thus contribute to osteosarcoma tumorigenesis. PMID- 12550768 TI - Allele-specific replication associated with aneuploidy in blood cells of patients with hematologic malignancies. AB - We hypothesize that coordination between the two DNA parental sets in somatic cells is essential for the stability of the diploid genome, and that its disruption is associated with the many alterations observed in the various cancerous phenotypes. As coordination between two allelic counterparts is well exemplified by synchrony in replication timing, we examined, in blood cells of patients suffering from various hematologic malignancies, replication patterns of five loci. These loci were three cancer-implicated genes (TP53, AML1, and RB1) and two nontranscribed sequences engaged in chromosome segregation. All five loci normally display synchrony in allelic replication timing. In addition, in order to exemplify an asynchronous mode of allelic replication, we followed the replication of allelic counterparts of an imprinted gene (SNRPN), which is distinguished by its asynchronous mode of allelic replication (allele-specific replication). Allelic replication patterns were studied by fluorescence in situ hybridization (FISH), which has been shown to distinguish between nonreplicated and replicated regions of the genome in interphase cells, based on the structure of the specific hybridization signals that are being detected. Using the FISH replication assay we observed, for all loci which normally exhibit synchrony in allelic replication, loss of synchrony when present in blood cells of patients with hematologic malignancies. The loss of synchrony in allelic replication in patients' cells was accompanied by aneuploidy (chromosome losses and gains), the hallmark of cancer. We were able to reinstate the normal pattern of replication in the patients' cells by introducing an inhibitor of DNA methylation. It thus appears loss of allelic coordination is an epigenetic alteration characterizing cancer, which is easily identified by simple cytogenetic means and has a potential use in both cancer investigation and detection. PMID- 12550769 TI - RARA fluorescence in situ hybridization overcomes the drawback of PML/RARA fluorescence in situ hybridization in follow-up of acute promyelocytic leukemia. AB - Determination of the remission of acute promyelocytic leukemia (APL) after chemotherapy can be difficult because many cases of APL show reverse transcription polymerase chain reaction positivity after consolidation treatment. Moreover, the discrimination of leukemic promyelocytes and regenerating promyelocytes by morphology is sometimes difficult. Although PML/RARA fluorescence in situ hybridization (FISH) can help, the major drawback of FISH is its high false positive rate, which reaches up to 5-10%. We used RARA FISH at the initial diagnosis (16 cases) and follow-up of APL patients (21 cases) with t(15;17), though RARA FISH was originally designed to detect translocations involving the RARA gene rather than t(15;17), and compared the results with those of PML/RARA FISH. A reference range for PML/RARA and RARA FISH was set using 50 normal control specimens. Using a RARA split probe, we were able to lower the reference range for RARA rearrangement down to 1.5%, which is significantly lower than that of PML/RARA (8%). Actually 74.2% (46/62 cases) of cases with positive signals of the PML/RARA rearrangement by the PML/RARA probe, showed absolutely negative results with the RARA split probe. By conducting RARA FISH, we were able to significantly resolve the difficulty in interpreting results around cut-off value in PML/RARA FISH. In conclusion, we believe that once the PML/RARA rearrangement is confirmed either by G-banding or FISH, RARA FISH is more effective than PML/RARA during the follow-up of APL after treatment. PMID- 12550770 TI - Chromosome 8 genetic analysis and phenotypic characterization of 21 ovarian cancer cell lines. AB - The short arm of chromosome 8 undergoes frequent loss of heterozygosity (LOH) in ovarian adenocarcinomas. Fine mapping has identified several distinct critical regions within 8p which undergo rates of LOH of 50% or greater, suggesting that there may be more than one tumor suppressor gene located on this chromosome arm. In an effort to refine the location of these putative tumor suppressor genes by homozygosity-mapping-of-deletion analysis, we have analyzed 21 ovarian cancer cell lines with 19 polymorphic microsatellite markers from 8p. Eleven of the cell lines (55%) were homozygous at every marker, indicating loss of an entire 8p arm. No smaller extended regions of hemizygosity were identified. Refinement of these 8p target regions was therefore not possible, but this analysis did identify the ovarian cancer cell lines that would be most appropriate for microcell-mediated chromosome transfer to complement the hypothesized mutation in the target tumor suppressor gene(s) on 8p. The 11 cell lines that had undergone 8p LOH were therefore characterized for colony formation in soft agar and tumor formation in nude mice. We identified four cell lines (JAM, OVCA4, OVCA5, and OVCA8) that were hemizygous for 8p and that formed colonies in soft agar and tumors in nude mice, making them ideal cell lines for chromosome 8 or candidate gene transfer. PMID- 12550771 TI - Profiling of differential expression of messenger RNA in normal, benign, and metastatic prostate cell lines. AB - To understand the phenotypic changes associated with prostate cancer development and metastasis, we investigated differential gene expression in primary and established prostate cell lines used as models. We have used a differential display of messenger RNA (DDRT-PCR) technique using 168 primer combinations and total RNA from BPH-1, LNCaP, and PC3 cells to identify filter-based cDNA microarrays containing 18,376 nonredundant clones of genes and expressed sequence tags (EST) using mRNA from PrEC and MDAPCa2a cells to identify genes that are differentially expressed in normal, benign, and cancerous prostate cell lines. Twenty-five cDNA with a significant difference in expression of 76 candidate cDNA, as identified by DDRT-PCR and confirmed by slot-blot analysis, were selected for sequence analysis. Of these, 14 cDNA were further confirmed by Northern blot analysis. Analysis of the cDNA microarray data showed that a variety of genes/EST were up- or down-regulated in the metastatic prostate tumor cells and a majority of these genes encode cytoskeletal proteins and proteins with regulatory function. Expression profile of two EST was confirmed by reverse transcription polymerase chain reaction. We also have identified a number of genes exhibiting differential expression in prostate cancer cells, which were not known earlier to be involved in prostate cancer. This report provides a comparative analysis of differential gene expression between normal prostatic epithelial cells and prostate cancer cells, and a foundation to facilitate in depth studies on the mechanism of prostate cancer development and metastasis. PMID- 12550772 TI - Application of multicolor fluorescence in situ hybridization analysis for detection of cross-contamination and in vitro progression in commonly used murine tumor cell lines. AB - Murine tumor models are potent tools for cancer studies, most of which make use of a limited number of murine tumor cell lines that are exchanged by many research groups around the world. Although cross-contamination and in vitro karyotypic progression are well-known risks with respect to the identity of tumor cell lines, these parameters are rarely evaluated. Notably, routine karyotyping of murine cell lines is laborious and technically demanding because mouse chromosomes are morphologically similar. We therefore used a 21-color fluorescence in situ hybridization (FISH) approach (COBRA) for screening two groups of frequently used murine tumor cell lines, each of which shares known immunologic determinants. Multicolor analysis revealed that the sharing of immunologic determinants among three murine lymphoma cell lines (EL-4, MBL-2, and RBL-5) is directly related to their common origin. In several of the cell lines, the chromosomal derivatives had rearranged further, suggesting that the cross contamination events were not recent. In contrast, karyotypic analysis of three murine colon cancer cell lines (C26, CC36, and C51) showed that these constituted independent tumor clones despite the sharing of immunologic determinants. Our data point out that cross-contamination and in vitro evolution of murine tumor cell lines are a common phenomenon, and that multicolor FISH analysis is an efficient tool for verifying the origin and tracking the evolution of murine cell lines. PMID- 12550773 TI - Simple technique of codenaturation and fluorescence in situ hybridization with probes in a microcamera. AB - We describe a rapid, precise and economical method of performing fluorescence in situ hybridization with probes in a microcamera, without chemical risk for the operator caused by the use of formamide. The application has been tried on metaphase spreads and interphase nuclei from peripheral blood or bone marrow cultures and on formalin-fixed, paraffin-embedded tissue. PMID- 12550774 TI - Clonal rearrangement of 15p11.2, 16p11.2, and 16p13.3 in a case of nodular fasciitis: additional evidence favoring nodular fasciitis as a benign neoplasm and not a reactive tumefaction. AB - This article describes a case of nodular fasciitis with the karyotype 47,XY,+4/46,XY,add(15)(p11.2), t(16;16)(p13.3;p11.2). The presence of clonal chromosomal abnormalities in this case, as well as in three previously reported cases, indicates that nodular fasciitis is a benign neoplasm and not a reactive lesion. PMID- 12550775 TI - Chromosomes, genes, and cancer breakpoints. PMID- 12550776 TI - Periodontal delivery of ipriflavone: new chitosan/PLGA film delivery system for a lipophilic drug. AB - The aim of the present work was to design a film dosage form for sustained delivery of ipriflavone into the periodontal pocket. For this purpose, monolayer composite systems made of ipriflavone loaded poly(D,L-lactide-co-glycolide) (PLGA) micromatrices in a chitosan film form, were obtained by emulsification/casting/evaporation technique. Multilayer films, made of three layers of polymers (chitosan/PLGA/chitosan), were also prepared and compared to monolayer films for their "in vitro" characteristics. Morphology and physico chemical properties of the different systems were evaluated. The influence of pH, ionic strength and enzymatic activity on film degradation, was also investigated. Significant differences in swelling, degradation and drug release were highlighted, depending on film structure and composition. In vitro experiments demonstrated that the composite micromatricial films represent a suitable dosage form to prolong ipriflavone release for 20 days. PMID- 12550777 TI - Power consumption profile analysis and tensile strength measurements during moist agglomeration. AB - This study was performed to elucidate the influences of process and formulation design on the granulation process using power consumption and tensile strength measurements. In order to record and analyze the power consumption profile "in process" a computer program was developed to be used for optimal end-point control in reproducible granule production. The program analyzes and calculates a characteristical point, the turning point of the S-shaped ascent of the profile. The tensile strength expresses the cohesiveness between the powder particles, which is dependent on saturation and capillary pressure. In order to investigate the influence of the amount of liquid present in the granular material on tensile strength a device was developed. The maxima of tensile strength occurred at 90% saturation, whereas the maxima of power consumption were determined at 100% saturation. The measured tensile strength sigma (N/m(2)) equals to the volume specific cohesion (J/m(3)). The present work proved that the power consumption measurement is an alternative, simple and inexpensive method to determine the cohesion of powder particles. The turning point is introduced as a signature of the starting material and furthermore as a parameter for the cohesiveness of the starting material and therefore for optimal end-point detection at an early stage. PMID- 12550778 TI - Evaluating dissolution profiles of an anti-HIV agent using ANOVA and non-linear regression models in JMP software. AB - A powerful statistical method was designed using JMP software to detect factors contributing to differences in the dissolution process of an antiviral drug delivered in an oral dosage form. Due to the large number of dissolution media available for solid dosage forms, a statistical method to choose the appropriate medium is critical for testing solid dosage forms. We have developed an analysis of variance model to analyze the overall dissolution profile obtained from the various media. In vitro tests were performed using a standard USP basket apparatus (Vankel Inc., Cary, NC), and the analysis used the restricted/residual maximum likelihood method (JMP software) to partition the variance due to media (pH 1.2 and 6.8, +SDS, water alone and at pH 1.2 with pepsin), time (repeated measure) and capsule (random effect). This allowed correct standard error estimates to be used to compare dissolution in different media using planned linear contrasts. The model provided us with statistically powerful criteria to identify significant differences in capsule dissolution across time and to quantify capsule-to-capsule population variance estimate. The time specific linear contrasts showed the largest sum of square values (SS) occurred at 180 min (SS=0.268) for the simulated SIF (pH 6.8) versus SGF (pH 1.2) comparison (DF=166, MSE=3.92 x 10(-3)). The dissolution processes were further characterized using a non-linear regression fit of a power law function to the data for each capsule. This resulted in a method to statistically differentiate between the dissolution processes of the capsules in different media. PMID- 12550779 TI - Spray-dried chitinosans. Part I: preparation and characterization. AB - PURPOSE: Physicochemical and micromeritic characterization of chitinosans. METHODS: Chitinosans subjected to N-deacetylation and depolymerization were characterized for degree of N-deacetylation (DD), molecular weight (MW), pK(a), particle size determination and morphology, tap/bulk density measurements, surface area determinations, and determination of flow properties. RESULTS: The chitinosan DDs and MWs were dependent on the processing conditions and ranged from 66 to 89% and 2-522 kDa, respectively. Chitinosan particle sizes and shapes were dependent on drying conditions (range 8-465 microm). Spray-dried chitinosans were spherical and had smaller particle sizes than the non-spray-dried materials which were irregularly shaped particles. Higher density values were obtained for processed materials than those for the raw material. Lower specific surface areas were observed for non-spray-dried chitinosans (0.28-1.59 m(2)/g) than for spray dried chitinosans (0.74-3.01 m(2)/g). Weight variation of chitinosan tablets indicated that spray-dried chitinosans possessed improved flow characteristics as compared with tray-dried chitinosans. CONCLUSIONS: The effect of drying method employed in chitinosan manufacture, i.e. spray versus tray drying, on the physicochemical and micromeritic properties of the resultant chitinosans were evaluated. Although the drying methods did not significantly influence the physicochemical properties, they affected the micromeritic properties of the resultant chitinosans. PMID- 12550780 TI - Spray-dried chitinosans. Part II: in vitro drug release from tablets made from spray-dried chitinosans. AB - PURPOSE: Application of spray-dried chitinosans as excipients for use in drug delivery systems was explored. METHODS: Spray- and tray-dried chitinosans previously N-deacetylated and depolymerized were used. Directly compressed tablets (200mg) containing tetracycline, chitinosan, and magnesium stearate were prepared. The tablets were characterized for dimensions, weight, friability, crushing strengths, disintegration, and dissolution. RESULTS: The tablet weights, thickness, and diameters were not affected by the chitinosan selected (P>0.05). Friability of tablets containing tray-dried chitinosans was generally higher (and crushing strengths were lower) than tablets containing spray-dried chitinosans. Chitinosan molecular weight, degree of N-deacetylation, and drying method used, significantly affected crushing strengths (P<0.0001). Disintegration times were affected only by the type of chitinosan (P<0.0001) but not by the drying method used (P>0.9). Dissolution from tablets was significantly affected by the chitinosan type (P<0.025), but not affected by the drying method (P>0.5). CONCLUSIONS: Spray drying improved binding functionality of chitinosans, thereby enhancing the tablet crushing strength; however, friability, disintegration, and dissolution profiles were not significantly affected. The data obtained from this study support the usefulness of spray-dried chitinosans as excipients for use in drug delivery systems. PMID- 12550781 TI - The usefulness of sugar surfactants as solubilizing agents in parenteral formulations. AB - The usefulness of sugar surfactants as solubilizing agents was assessed and compared to commercial polyoxyethylene-based surfactants. The sugar surfactants examined comprised of monosaccharides or disaccharides with alkyl chains ranging from C(8) to C(12). Each surfactant was investigated with respect to solubilization capacity for felodipine and haemolytic activity. The haemolytic activity was determined using a static method in which surfactant solutions were added to fresh dog blood. The polyoxyethylene-based surfactants were found to be more suitable as solubilizing agents than the sugar surfactants due to better solubilization capacities combined with lower haemolytic activities. The sugar surfactants caused severe haemolysis below or at the critical micelle concentration, in contrast to the polyoxyethylene-based surfactants that are nonhaemolytic in this concentration range. The structure-related variations in haemolytic activity are probably due to variations in the surfactants partition coefficients for the distribution equilibrium between the aqueous phase and the cell membrane. Longer alkyl chains cause higher haemolytic activity, while larger saccharide groups lower the activity. The clear difference between sugar and polyoxyethylene surfactants, which are considerably less haemolytic, is due to a combination of low critical micelle concentrations and presumably low degrees of partitioning of the latter surfactants into the cell membranes. PMID- 12550783 TI - Design of pH-independent controlled release matrix tablets for acidic drugs. AB - The rate and extent of drug release from most controlled release systems are influenced by the pH of the dissolution medium for drugs with pH-dependent solubility. This dependency of drug release on pH may lead to additional inter- and intra-subject variability in drug absorption. In the present study, a pH independent controlled release matrix system for acidic drugs was designed by incorporating release-modifiers in the formulation. Controlled release matrix tablets were prepared by compression of divalproex sodium, Methocel K4M and Eudragit E 100 or Fujicalin as the release-modifier. For formulations without any release-modifier, the extent and rate of drug release at pH 6.8 was much higher than that at pH 1.0. Formulations containing Eudragit E 100 provided drug release that was essentially independent of pH. This was achieved because Eudragit E 100 significantly increased the drug release in acidic medium and slightly decreased the release rate at higher pH. The increased release in the acidic medium can be attributed to the elevation of the micro-environmental pH in the swollen polymer gel layer. Formulations containing Fujicalin were less effective than those containing Eudragit E 100. This was attributed to the relative inability to elevate the pH and shorter residence time of Fujicalin in the matrix relative to Eudragit E 100. PMID- 12550782 TI - Preparation and pharmacokinetics of pirarubicin loaded dehydration-rehydration vesicles. AB - Liposomes entrapped pirarubicin (THP, L-THP) were prepared by the modified dehydration-rehydration vesicle (DRV) method, and their pharmacokinetics and antitumor effects were evaluated in mice bearing M5076 liver metastasis tumor. After small unilamellar vesicles (SUVs) composed of egg lecithin, cholesterol (Ch), beta-sitosterol beta-D-glucoside (Sit-G) and oleic acid (OA) were freeze dried with THP and sugars, rehydration of the lyophilized powders led to form the larger vesicles entrapping drugs, but the proper amounts of sugars and OA to lipids (sucrose/lipid=8 (w/w)) maintained the small particle size (about 340nm) with high entrapment (80.7%) of THP. After intravenous injection of L-THP, the accumulations of THP in the liver and heart were approximately 4-fold higher and half lower, respectively, than those of free THP (F-THP). L-THP had superior antitumor effect in 10mg/kg intravenous administration without significant body weight loss. L-THP is a potential drug dosage form of liver cancer treatment since the liposomes carry THP to the liver. PMID- 12550784 TI - Influence of physico-chemical carrier properties on the in vitro aerosol deposition from interactive mixtures. AB - Interactive mixtures were prepared containing 5% (w/w) salbutamol sulfate using various lactose carrier systems, including sieved fractions and blended mixtures of coarse and fine particles. The solid state and powder properties of the lactose carriers were examined by laser diffraction, differential scanning calorimetry, thermogravimetric analysis, powder X-ray diffraction, vapor sorption gravimetry, rotating drum and atomic force microscopy. The in vitro aerosol deposition was determined using a twin-stage impinger with a Rotahaler at an airflow rate of 60l/min. The fine particle fraction (FPF) of salbutamol sulfate was determined using a validated HPLC assay. All samples were highly crystalline with minimal moisture sorption and the major phase in all samples was alpha lactose monohydrate. Significant differences in FPF were observed using the various carrier systems. FPF increased with decreasing carrier d(50%) (r(2)=0.919) and increasing proportion of fine carrier particles (below 5 microm) (r(2)=0.841). Carriers consisting of very large proportions of fine particles showed low FPF and did not fit the correlation. The presence of coarse carrier particle fractions was essential to achieve maximum FPF, which occurred when about 10% of fine carrier particles were present in the mixture. Dispersion characteristics may be related to the degree of drug aggregation on the carrier surface. PMID- 12550785 TI - The effect of the drug/polymer ratio on the properties of the verapamil HCl loaded microspheres. AB - In this study, the microspheres containing verapamil hydrochloride (VRP) were prepared with Eudragit RS 100 by solvent evaporation method. In the solvent evaporation method one of the parameters which affect to the formation and properties of the microspheres is the variations of drug/polymer ratios. The aim of our study is to examine the effects of this parameter on the VRP loaded microspheres. To achieve this purpose, only drug/polymer ratio was altered while the other formulation parameters were kept constant and percentage yield value, incorporation efficiency, particle size and distribution of the microspheres were analyzed and micrographs of the microspheres were taken to determine the effects of the increase in the polymer amount of formulations. All the dispersed phase viscosities were evaluated by comparing them with the variations in particle size and distribution of the microspheres. In vitro dissolution tests were done by using dissolution media with three different pH in sequence as half-change method with flow through cell and the effect of the variation in polymer ratio on drug dissolution was evaluated according to dissolution test results. As a result of our study, it is thought that the variation in drug/polymer ratios might have an influence on the physical characteristics of the microspheres and the increasing amount of polymer might be result in decreased drug dissolve. PMID- 12550786 TI - Stabilization of recombinant interferon-alpha by pegylation for encapsulation in PLGA microspheres. AB - Interferon-alpha (IFN) was pegylated and encapsulated in biodegradable microspheres to achieve a long-acting formulation. IFN was pegylated with methoxy polyethylene glycol (mPEG, MW 2000 or 5000). The conjugation procedures were optimized in terms of concentration of the reactants and the pH condition of the medium. The conjugates (IFN-mPEG(2000) and IFN-mPEG(5000)) were characterized using SDS-PAGE, size-exclusion-HPLC (SE-HPLC) and matrix-assisted laser desorption and ionization time-of-flight (MALDI-TOF) mass spectroscopy. The optimized IFN-mPEG conjugates consisted of mono- and multi-pegylated derivatives along with a small amount of native IFN (soybean oil emulsion>linseed oil emulsion>DOTAP liposome. The squalene emulsion showed the least cytotoxicity with or without serum. For in vivo gene transfer, the squalene emulsion also had the most potent transfection activity in the mouse after intravenous administration. Squalene as the oil component can enhance the stability of cationic emulsion more effectively that could be useful for the transfer of genes in vitro and in vivo. PMID- 12550801 TI - Determining the compression behaviour of pharmaceutical powders from the force distance compression profile. AB - The force-displacement curves obtained from an eccentric tablet machine were examined in a new way. The tendency of the material for plastic deformation, fragmentation and elasticity is expressed as numerical values, which are comparable between different materials. The dependence of these numerical values on the compression pressure was modelled. The accuracy of the displacement measurement was improved by filtering out noise from the measurement data by a novel method. The plastic deformation of the material near the force maximum of the compression cycle could be seen accurately from this precise displacement data. The elastic deformation of the tablet machine was also defined very precisely from the running machine. Tablets were made with an eccentric tablet machine using fixed lower and upper punch adjustments. This ensured that the speed of the upper punch and the theoretical height of the tablets were the same for all compactions. Therefore, only the properties of the materials determined the differences in the shape of the compression curves. The test materials used were alpha-lactose monohydrate, two grades of microcrystalline cellulose, maize starch and dicalcium phosphate dihydrate. The results showed that the use of accurate displacement measurement is essential in order, to see the small variations in the shape of the force-displacement curve near the force maximum of the compression cycle, and it made it possible to dynamically calibrate the elastic deformation of the eccentric tablet machine during compression. It turned out that the numerical values obtained with the new method described the plastic, brittle and elastic properties of the tested materials satisfactorily in a wide compression pressure range. PMID- 12550802 TI - The influence of surface modification on the cytotoxicity of PAMAM dendrimers. AB - The influence of surface modification on the cytotoxicity of PAMAM dendrimers was examined using Caco-2 cells. Dendrimers were modified by conjugating either lauroyl chains or polyethylene glycol (PEG) 2000 onto the surface of cationic PAMAM dendrimers (G2, G3, G4). The cytotoxicity of unmodified dendrimers towards Caco-2 cells was appreciably higher for cationic (whole generation) compared with anionic (half generation) dendrimers and for both types increased with increasing size (generation) and concentration. A marked decrease in the cytotoxicity of cationic PAMAM dendrimers was noted when the surface was modified, with the addition of six lauroyl or four PEG chains being particularly effective in decreasing cytotoxicity. This decrease in cytotoxicity is thought to be due to a reduction/shielding of the positive charge on the dendrimer surface by the attached chains. The cytotoxicity of dendrimer-based delivery systems is likely to be very different from the parent dendrimer. PMID- 12550803 TI - Light scattering investigations on freeze-dried glucocorticoids in aqueous solution. AB - Aqueous solutions of three glucocorticoid drug substances were investigated using dynamic light scattering. Evidence of micelle formation was found. The identified diffusion coefficients, micellar radii and the dependence of these parameters on the concentration of the solution as well as on the molecular structure of the drugs are discussed. It is postulated that the micellar character of the solutions is one reason for the ease of lyophilization of such molecules without additional substances. PMID- 12550804 TI - Hypoglycemic effect of surfactant-coated insulin solubilized in a novel solid-in oil-in-water (S/O/W) emulsion. AB - A novel solid-in-oil-in-water (S/O/W) emulsion for oral administration of insulin has been developed using surfactant-coated insulin. The S/O/W emulsion prepared by a shirasu porous glass (SPG) membrane provided a sharp size distribution and was stable. Leakage of insulin from the S/O/W emulsions was not observed for several days. The S/O/W emulsion showed the hypoglycemic activity for a long period after oral administration to rats. PMID- 12550805 TI - In Check Dial: accuracy for Diskus and Turbuhaler. AB - OBJECTIVE: The In Check Dial was developed to evaluate whether a patient is able to generate an adequate peak inspiratory flow (PIF) through a certain inhalation device. The inhalation profile recorder (IPR) is a calibrated instrument that measures flows through Diskus and Turbuhaler in our research setting. The aim of this study was to compare the PIFs of patients when inhaling through a Diskus or Turbuhaler connected with the IPR (PIF_diskus and PIF_TH) to the flows through the corresponding orifices of the In Check Dial (Diskus_In Check and TH_In Check). METHODS: Twenty-four stable asthma and twenty-one chronic obstructive pulmonary disease (COPD) patients inhaled, on two separate occasions, in randomised order, via the Diskus or Turbuhaler connected with the IPR. Subsequently, patients inhaled through the In Check Dial using the orifices of Diskus or Turbuhaler. RESULTS: The difference between Diskus_In Check and PIF_diskus was 3.9 (11.9)l/min (P=0.038). The difference between TH_In Check and PIF_TH was 3.5 (10.6) l/min (NS). All Diskus- and Turbuhaler-inhalations were performed with the minimum required flow of 30 l/min. However, four COPD patients inhaled with the non-optimal flow (<60 l/min) through the Turbuhaler. The In Check Dial did not indicate two of them. CONCLUSION: Measuring PIF through Diskus and Turbuhaler using the IPR and the In Check Dial, respectively shows a disagreement of 3.9l/min. A disagreement of 3.5l/min was found for the Turbuhaler. The In Check Dial did not identify two of four patients as 'non optimal' users. PMID- 12550806 TI - Defining the Neutral Zone of sheep intervertebral joints during dynamic motions: an in vitro study. AB - OBJECTIVE: To make an experimental assessment of the Neutral Zone of intervertebral joints during dynamic spinal motion in flexion/extension, lateral bending and axial rotation and to develop a criterion for its definition. DESIGN: Dynamic mechanical testing of sheep intervertebral joints with a six-degree of freedom robotic facility under position control. BACKGROUND: The Neutral Zone is defined as a region of no or little resistance to motion in the middle of an intervertebral joint's range of movement. Previous studies have used quasi-static loading regimes that do not model physiological activity. This study simulated physiological movements using a robotic testing facility to address this issue. METHODS: Five spines from mature sheep were used and three motion segments were tested from each spine. The robotic facility enabled the testing regime to be defined for each individual joint based on its geometry. The joints were tested by cycling through the full range of physiological movement in flexion/extension, lateral bending and axial rotation. RESULTS: A Neutral Zone was found to exist during dynamic movements only in flexion/extension. The results suggested that a Neutral Zone does not exist in lateral bending or axial rotation. The zygapophysial joints were shown to be significant in determining the mechanics of the intervertebral joints as their removal increased the Neutral Zone in all cases. A new criterion for defining the size of the Neutral Zone during dynamic motion was proposed and its implications for spinal movements in life discussed. CONCLUSIONS: A Neutral Zone exists in flexion/extension during dynamic movements of intervertebral joints and is a feature of the natural range of joint motion. This has important implications for the muscular control of the spine consisting of several intrinsically lax joints stacked on one another. RELEVANCE: The existence of a Neutral Zone is a feature of the natural range of joint motion and requires complex control of intervertebral joints by the spinal muscles. Defining the biomechanical response throughout the physiological range of motion (RoM) is important in understanding possible injury and rehabilitation mechanisms. PMID- 12550807 TI - The iliolumbar ligament: its influence on stability of the sacroiliac joint. AB - STUDY DESIGN: In human specimens the influence of the iliolumbar ligament on sacroiliac joint stability was tested during incremental moments applied to the sacroiliac joints. OBJECTIVES: To assess whether the iliolumbar ligament is able to restrict sacroiliac joint mobility in embalmed cadavers. BACKGROUND: Firstly, the sacroiliac joint can play an important role in non-specific low back pain; hence, its mobility and stability are of special interest. Secondly, the iliolumbar ligament is considered to be an important source of chronic low back pain. Data on a functional relation between the iliolumbar ligament and sacroiliac joint mobility are lacking. METHODS: In 12 human specimens an incremental moment was applied to the sacroiliac joint to induce rotation in the sagittal plane. After the assessment of the relationship between rotation angle and moment in the intact situation, specific parts of the iliolumbar ligaments were transected. After each partial transection the measurements were repeated. RESULTS: Sacroiliac joint mobility in the sagittal plane was significantly increased after a total cut of both iliolumbar ligaments. This increase was in particular due to the transection of a specific part of the iliolumbar ligament, the ventral band. CONCLUSIONS: The main conclusions are: (a) the iliolumbar ligaments restrict sacroiliac joint sagittal mobility; (b) the ventral band of the iliolumbar ligament contributes most to this restriction. RELEVANCE: In embalmed human cadavers, the mobility of the sacroiliac joint increases after sequential cutting of specific parts of the iliolumbar ligaments. It can be expected that severance of this ligament during surgery will lead to increase of mobility and hence loss of stability of the sacroiliac joint. As a consequence adjacent structures will be affected. This may well be a cause of pain in patients with failed back surgery. PMID- 12550808 TI - Mechanical energy and power flow of the upper extremity in manual wheelchair propulsion. AB - OBJECTIVE: To investigate the characteristics of mechanical energy and power flow of the upper limb during wheelchair propulsion. DESIGN: Mechanical energy and power flow of segments were calculated. BACKGROUND: Very few studies have taken into account the mechanical energy and power flow of the musculoskeletal system during wheelchair propulsion. Mechanical energy and power flow have proven to be useful tools for investigating locomotion disorders during human gait. METHODS: Twelve healthy male adults (mean age, 23.5 years) were recruited for this study. Three-dimensional kinematic and kinetic data of the upper extremity were collected during wheelchair propulsion using a Hi-Res Expert Vision system and an instrumented wheel, respectively. RESULTS: During the initiation of the propulsion phase, joint power is generated in the upper arm or is transferred from the trunk downward to the forearm and hand to propel the wheel forward. During terminal propulsion, joint power is transferred upward to the trunk from the forearm and upper arm. The rate of change of mechanical energy and power flow for the forearm and hand have similar patterns, but the upper arm values differ. CONCLUSIONS: Joint power plays an important role in energy transfer as well as the energy generated and absorbed by muscles spanning the joints during wheelchair propulsion. RELEVANCE: Energy and power flow information during wheelchair propulsion allows us to gain a better understanding of the coordination of the movement by the musculoskeletal system. PMID- 12550809 TI - The effect of metallic radial head arthroplasty on radiocapitellar joint contact area. AB - OBJECTIVE: This study determined the effect of metallic radial head arthroplasty on radiocapitellar joint contact area. DESIGN: The contact area of eight intact radiocapitellar joints was assessed during simulated compressive loading. Subsequently, the same experiments were repeated after reconstruction with three different radial head implant sizes. BACKGROUND: There has been an increasing use of metal radial head implants relative to silicone implants. However, the contact characteristics with the metal reconstruction have not been determined. METHODS: A 100 N compressive load was applied to the radiocapitellar joint at three different flexion angles for the native joint and three different sizes of a metal head implant. An impression material was employed to quantify joint contact area. RESULTS: Following metallic radial head arthroplasty, the contact area decreased by approximately two-thirds relative to the native radiocapitellar joint (P<0.001). Smaller implant sizes resulted in slightly larger contact areas (P<0.05). In addition, contact area decreased as joint flexion angle increased (P<0.04). CONCLUSIONS: Selection of radial head implant size is likely not a significant factor with regard to contact area mechanics of the radiocapitellar articulation. RELEVANCE: The effect of radial head implant size on the contact area produced with the native capitellum is not known. This study shows that the design or selection of a metallic radial head implant from the viewpoint of size is likely not an important factor with regard to minimizing joint contact stresses. PMID- 12550810 TI - Intrinsic stability of an unconstrained metacarpophalangeal joint implant. AB - OBJECTIVE: To compare the intrinsic stability of an unconstrained resurfacing metacarpophalangeal arthroplasty to that of a normal human cadaveric joint. DESIGN: Cadaveric joints and metacarpophalangeal prostheses were studied in a mechanical testing machine at different angles and axial loads to determine the stability ratio in eight directions of movement. BACKGROUND: An unconstrained resurfacing arthroplasty was designed to replicate the normal anatomy with the exception of the proximal component having a greater arc of curvature on its dorsal aspect. METHODS: Eight fresh-frozen cadaveric joints and five different sizes of the AVANTA metacarpophalangeal prosthesis were studied at 0 degrees, 45 degrees and 90 degrees angles of flexion and at eight different directions of motion with three different axial loads (0, 20, 40 N). A 6-component load cell measured the force needed to sublux the joint. The stability ratio was the measured outcome and is defined as ratio of the force of subluxation to the axial force. RESULTS AND CONCLUSIONS: The unconstrained resurfacing arthroplasty has more intrinsic stability than the cadaveric metacarpophalangeal joint in all eight directions tested. RELEVANCE: A major complication of metacarpophalangeal implants is ulnopalmar subluxation. The AVANTA implant is designed to decrease the risk of ulnopalmar subluxation by having a greater arc of curvature on the dorsal aspect of the proximal component. This study shows that the designed implant has greater stability due to the geometry of the implant compared to that of the anatomical joint. PMID- 12550811 TI - The accuracy of estimating proximal tibial translation during natural cadence walking: bone vs. skin mounted targets. AB - OBJECTIVE: To assess the efficacy of estimating proximal tibial translation using video-based motion capture and an array of surface-mounted targets ideal for tracking motion of the tibia. DESIGN: Superficial and bone-anchored tracking targets were used to create two independent sets of data locating the proximal tibia in a global coordinate system. BACKGROUND: Knowledge of the effect that soft tissue movement has on estimates of proximal tibial translation has not been reported to date. This basic information is necessary to establish construct validity for any study proposing to document tibial translation using standard video-based motion capture methods. METHODS: A six camera motion capture system was used to collect surface-mounted and bone-anchored data for seven healthy adult subjects walking at a self-selected speed. The subjects walked along the positive Y-axis of the global reference system, with the positive Z-axis oriented vertically. RESULTS: Average peak differences in the location of the proximal tibia calculated from the bone and surface-mounted targets were 7.1, 3.7 and 2.1 mm along the X, Y and Z axes of the global coordinate system respectively. Individual subject peak differences were as large as 14.1, 11.8 and 8.3 mm along the X, Y and Z axes. CONCLUSIONS: Estimates of tibial translation with a measurement resolution better than 3 mm are not likely using standard motion capture methods and tracking targets attached superficially to the lower leg. RELEVANCE: The results of this study clearly depict the considerable effect that soft tissue motion of the lower leg has on estimates of proximal tibial translation. Without consideration for the difficulties in measuring femoral, or patellar motion, we believe it is not feasible to routinely obtain sufficiently accurate estimates of detailed knee joint translations using superficial tracking target attachment methods. PMID- 12550813 TI - A clinically applicable EMG-force model to quantify active stabilization of the knee after a lesion of the anterior cruciate ligament. AB - OBJECTIVES: To investigate whether a simple electromyography-force (EMG-force) model can be used to measure different levels of co-contraction about the knee for healthy subjects and patients with an anterior cruciate ligament deficiency. DESIGN: To evaluate an EMG-to-force processing model, two groups of subjects, with and without deficiency of the anterior cruciate ligament, participated in experiments in which surface EMG, kinematics and kinetics about the knee were recorded during isokinetic and functional movements. BACKGROUND: Clinical and biomechanical evidence supports the hypothesis that higher level of co contraction of quadriceps and hamstrings provide an active stabilization of the knee to compensate for the lost anterior cruciate ligament. To quantify the level of co-contraction, the contribution of both agonist and antagonist muscles to the net joint moment must be known. METHODS: Surface EMG levels were calibrated to moment by means of a limited number of isokinetic contractions about the knee. With these calibration values, an estimate of the muscle moments during a vertical jump were obtained and compared with the net joint moment, calculated with inverse dynamics. Also co-contraction indices were determined. RESULTS: The EMG-force model provided a fair estimate of the net joint moment. The co contraction index in anterior cruciate ligament deficient subjects was significantly higher (mean 0.54 (SD, 0.04)) compared to healthy subjects (mean 0.25 (SD, 0.07)). CONCLUSIONS: Although the EMG-to-force processing model is not perfectly accurate, it is appropriate within a clinical context. RELEVANCE: Previous research supports the hypothesis that subjects with an anterior cruciate ligament deficiency compensate the loss of passive stability by developing higher co-activation levels of the knee muscles, i.e. active stabilization. Quantifying co-contraction may serve as a valuable parameter to evaluate clinical interventions and rehabilitation processes. The EMG-force model presented in this study appears to be a useful instrument for this purpose. PMID- 12550812 TI - Gait perturbation response in chronic anterior cruciate ligament deficiency and repair. AB - OBJECTIVE: To determine how chronic anterior cruciate ligament deficient and surgically repaired subjects react to unexpected forward perturbations during gait as compared to healthy controls. DESIGN: Gait testing of 10 chronic anterior cruciate ligament deficient subjects prior to and three months following reconstructive surgery, and 10 uninjured controls. BACKGROUND: The ability of an anterior cruciate ligament injured individual to react and maintain equilibrium during gait perturbations is critical for the prevention of reinjury. No studies have investigated how these individuals respond to unexpected perturbations during normal gait. METHODS: An unexpected forward perturbation was induced upon heel strike using a force plate capable of translational movement. RESULTS: Prior to surgery, the anterior cruciate ligament subjects exhibited a greater knee extensor moment in response to the perturbation compared to healthy controls. Following surgery, the anterior cruciate ligament injured subjects exhibited a static knee position and a sustained knee extensor moment throughout stance in response to the perturbation as compared to controls. CONCLUSIONS: These data suggest that chronic anterior cruciate ligament deficient subjects rely heavily on knee extensor musculature to prevent collapse in response to an unexpected perturbation. This same reactive response was more pronounced 3 months following surgery. RELEVANCE: The results suggest that, prior to and following surgery, chronic anterior cruciate ligament injured subjects respond differently than healthy controls to an unexpected perturbation during gait. Anterior cruciate ligament injured or repaired subjects do not reduce or avoid vigorous contraction of the quadriceps muscles when responding to gait perturbations. PMID- 12550814 TI - Radiofrequency treatment weakens the fatigue characteristics of rabbit anterior cruciate ligament. AB - OBJECTIVE: To evaluate the radiofrequency treatment applied to an intact anterior cruciate ligament, and to quantify the mechanical effects of controlled cyclic loading (simulating activities of daily living) post-treatment. DESIGN: An in vitro radiofrequency energy application to the rabbit anterior cruciate ligament and cyclic loading of the treated ligament. BACKGROUND: Effect of cyclic loading on the radiofrequency treated ligament in a controlled environment is not known. METHODS: In the treatment group radiofrequency treatment was applied to an intact ligament and then the ligament was cyclically loaded for 3000 cycles, and stretched-to-failure. Control group was similar to treatment, except that the radiofrequency treatment was not applied. The cyclic loading (approximately 6% tensile strain) was kept below the injury threshold of the rabbit anterior cruciate ligament and was designed to simulate loading post-treatment. A relaxation test (5% constant strain for 180 s) was used to compare the results at different steps in the experiment; within each group and between the two groups. RESULTS: Relaxation force increased in the treatment group (average 108% of intact, P<0.05), but decreased after the cyclic loading (average 71% of intact, P<0.05). There were significant differences in the force-deformation-failure curves between the treatment and control groups: average treated ligament failed at 59% and 57% of the control values respectively for the deformation and force (P<0.05). CONCLUSIONS: Using radiofrequency treatment makes rabbit anterior cruciate ligament weaker following cyclic loading. RELEVANCE: The radiofrequency treatment weakens the anterior cruciate ligament, and therefore, it perhaps should be protected until healing and adaptation restore its full strength. PMID- 12550815 TI - Velocity dependent passive plantarflexor resistive torque in patients with acquired brain injury. AB - OBJECTIVES: This study sought to determine whether factors other than stretch reflex excitability contribute to velocity dependent passive plantarflexor resistive torque following brain injury. BACKGROUND: In patients with acquired brain injury increased resistance to passive muscle lengthening commonly results from abnormal muscle contraction, secondary to disinhibition of descending motor pathways, in addition to rheologic changes within the musculo-tendinous unit. Hyper-excitable tonic stretch reflex responses (spasticity) have traditionally been considered to be the main factor influencing resistance that is velocity dependent. METHODS: Ten adults with brain injury and eighteen age matched controls were studied. A computer controlled torque measurement system was utilised to evaluate resistance to dorsiflexion stretches at two velocities (5 degrees and 25 degrees s(-1)). Only stretches which did not evoke muscle contraction were included in the data analysis. The mean difference and 95% confidence limits in passive plantarflexor resistive torque at two stretch velocities, measured over a defined portion of the test movement, were compared between subject groups. RESULTS: A velocity dependent increase in passive plantarflexor resistive torque was evident when the ankle was dorsiflexed past the neutral position in both subjects with brain injury and controls. However, the mean difference was approximately 10 times greater in neurologically impaired limbs compared with control values. CONCLUSIONS: These data indicate that an important component of velocity dependent resistance to passive muscle lengthening in adults with brain injury can be mechanical, and unrelated to stretch induced reflex muscle contraction. RELEVANCE: Increased resistive torque during rapid muscle lengthening may represent a compensatory adaptation for reduced distal motor control following brain injury. A velocity dependent increase in passive plantarflexor resistive torque has the potential to improve stability during gait and provide mechanical resistance to sudden external perturbations. PMID- 12550816 TI - Stiffness characteristics and inter-fragmentary displacements with different hybrid external fixators. AB - OBJECTIVES: To determine how the different approaches of constructing a hybrid external skeletal fixator affect the mechanical environment at the fracture site. DESIGN: A comparative biomechanical analysis of external fixators that represent a spectrum of designs from unilateral to circular fixators. BACKGROUND: The most pertinent parameter that affects fracture healing is the inter-fragmentary displacement. Most studies on the mechanical performance of external fixators have concentrated on the overall stiffness of the fixators. METHODS: Mechanical testing was performed on four types of hybrid fixators in which the overall stiffness of the fixator as well as the axial, transverse shear and angular displacements at the fracture site were measured. RESULTS: The Ilizarov hybrid fixator with one wire and one screw on each ring behaved more like a unilateral fixator than a circular fixator. The bar-ring hybrid fixator that simply connected a unilateral fixator body to a wire/ring assembly was too flexible. Reinforcing the bar-ring hybrid system with diagonally placed struts eliminated the deformation associated with the unilateral body and the resulting construct performed considerably better. The two-ring hybrid fixator most closely resembled the Ilizarov fixator. CONCLUSIONS: Depending on the construction characteristics hybrid fixators possess a spectrum of mechanical characteristics of both unilateral and circular fixators. The constructions using two rings in which at least one bone segment is supported entirely by wires appears to have the best mechanical characteristics. RELEVANCE: Understanding how the different approaches of constructing a hybrid fixator affect the mechanical environment at the fracture site will enable surgeons to chose or build the most appropriate fixator for each clinical situation. PMID- 12550818 TI - Adolescent resilience: a concept analysis. AB - There is need for greater clarity around the concept of resilience as it relates to the period of adolescence. Literature on resilience published between 1990 and 2000 and relevant to adolescents aged between 12- and 18-years of age was reviewed with the aim of examining the various uses of the term, and commenting on how specific ways of conceptualizing of resilience may help develop new research agendas in the field. By bringing together ideas on resilience from a variety of research and clinical perspectives, the purpose of the review is to explicate core elements of resilience in more precise ways, in the hope that greater conceptual clarity will lead to a range of tailored interventions that benefit young people. PMID- 12550819 TI - Prosocial behaviours of young adolescents: a focus group study. AB - This study investigated young adolescents' perceptions of their peers' prosocial behaviours. In eight focus groups, 53 11- to 13-year olds described specific prosocial acts of their peers. Results suggest that traditional research has not addressed the diversity of prosocial behaviours that youth enact, nor emphasized behaviours that are salient to young adolescents. Such behaviours included standing up for others, encouraging others, helping others develop skills, including others who are left out, and being humorous. Facilitating emotional regulation of others emerged as an important component of prosocial behaviour. These data can help guide future research on prosocial development to include a broader array of authentic behaviours of young adolescents. PMID- 12550820 TI - The mental health of young people attending a Youth Offending Team: a descriptive study. AB - This study documents the health needs of young offenders aged 10-17 attending a community Youth Offending Team (YOT). All young people known to a YOT on a selected date were identified. Health information was collected via semi structured interviews and standardized questionnaires. Potential mental health problems that required further specialist assessment were identified in 56% of those assessed. Alcohol was consumed more than twice per week by 68%, with 47% having recently smoked cannabis, and, 11% recently using heroin, methadone or crack cocaine. Use of secondary health-care services was common although contact with primary-care services was less frequent with almost half having no contact with a GP in the past year. The process of meeting the physical and mental health needs of young offenders in the community is discussed. PMID- 12550821 TI - Female offenders referred for community-based mental health service as compared to other service-referred youth: correlates of conviction. AB - Data from a large federally supported national evaluation of system-of-care community mental health services were analysed to identify correlates of conviction. Female adolescents with a reported history of criminal conviction (n=88) were compared to three other service-referred youth groups: females without conviction histories (n=664), males with conviction histories (n=199), and males without conviction histories (n=1230) for possible differences in number and type of family, individual, and school-related life challenges. Multinomial regression analyses were first used to compare the quantity of child and family correlates in each conviction group, and then to test specific correlates in the individual, family, and school domains. The conditional odds of reporting a high vs. low number of child correlates was found to be significantly greater for females with a history of conviction compared to all other groups, over and above the number of family risk factors. Further, service-referred females with a conviction history, when compared to other service-referred youth groups, were much more likely to report having experienced a living instability (e.g. history of running away, multiple living arrangements) and personal adverse life events (e.g. history of drug and alcohol use, sexual abuse). Implications for community-based interventions and treatment are discussed. PMID- 12550822 TI - The role of father involvement in children's later mental health. AB - Data on 8441 cohort members of the National Child Development Study were used to explore links between father involvement at age 7 and emotional and behavioural problems at age 16, and between father involvement at age 16 and psychological distress at age 33, controlling for mother involvement and known confounds. Father involvement at age 7 protected against psychological maladjustment in adolescents from non-intact families, and father involvement at age 16 protected against adult psychological distress in women. There was no evidence suggesting that the impact of father involvement in adolescence on children's later mental health in adult life varies with the level of mother involvement. PMID- 12550823 TI - The effectiveness of individual and group-based parenting programmes in improving outcomes for teenage mothers and their children: a systematic review. AB - There is evidence from a range of studies showing adverse child outcomes for the children of teenage parents. Parenting programmes are increasingly being used to promote the well-being of parents and children, and this review aims to establish whether they can improve outcomes for teenage parents and their children. The findings of the review are based on 14 studies that used varying study designs, and are therefore limited. The results suggest, however, that parenting programmes can be effective in improving a range of psychosocial and developmental outcomes for teenage mothers and their children. Further research is needed. PMID- 12550824 TI - Escaping or connecting? Characteristics of youth who form close online relationships. AB - We used data from a US national sample of Internet users, ages 10-17 (N=1501), to explore the characteristics of youth who had formed close relationships with people they met on the Internet (n=210). Girls who had high levels of conflict with parents or were highly troubled were more likely than other girls to have close online relationships, as were boys who had low levels of communication with parents or were highly troubled, compared to other boys. Age, race and aspects of Internet use were also related. We know little about the nature or quality of the close online relationships, but youth with these sorts of problems may be more vulnerable to online exploitation and to other possible ill effects of online relationships. At the same time, these relationships may have helpful aspects. PMID- 12550825 TI - Illicit drug use, cigarette smoking and alcohol drinking behaviour among a sample of high school adolescents in the Pietersburg area of the Northern Province, South Africa. AB - AIM: The aim of this study was to investigate the prevalence of illicit drug use, cigarette smoking and alcohol drinking behaviour among a sample of high-school adolescents in the Pietersburg area (Central Region) of the Northern Province, South Africa. PARTICIPANTS: Participants in this study were 435 secondary-school adolescent students around the Pietersburg area (Central Region), in the Northern Province. Two hundred and forty-three (56%) participants were female and 192 (44%) were male. They were aged between 15 and 19 years, with the mean age of 17.25 years (S.D.=1.34). Their educational level ranged between standards 7 and 10 (Grades 9-12). INSTRUMENTS: Participants' demographic variables were determined by use of a questionnaire which covered participants personal characteristics such as age and gender, as well as educational level. Illicit drug use, cigarette smoking, and alcohol drinking behaviour were also measured using the questionnaire. RESULTS: Findings of this study indicate the prevalence rate of 19.8% for illicit drug use, 10.6% for cigarette smoking and 39.1% for alcohol consumption among the participants. Both drug use, cigarette smoking and alcohol consumption are associated more with males than with females. The majority of the drug users and cigarette smokers indicated that they do so when they are bored, tired or stressed up, or at parties; and most of those who drink alcohol indicated that they do so at parties, weekends, or any other time. The mean age for first drug use is 14.9 years (S.D.=1.77); 14.54 years (S.D.=1.80) for first cigarette smoking and 15.33 years (S.D.=1.91) for first alcohol consumption. Among those who drink alcohol, 24.1% and 14.1% experience unplanned drunkenness and irritability, respectively. Unplanned drunkenness is associated with males, while unplanned irritability (after drinking alcohol) is not associated with any gender. CONCLUSION: Mental health workers and the law enforcement agencies should take note of the above findings while planning preventive and therapeutic strategies for the reduction of drug use, cigarette smoking and alcohol use and abuse among adolescents in the area. PMID- 12551642 TI - Intraocular lens power accuracy: how are we doing? PMID- 12550826 TI - The preliminary development of a new self-report measure for OCD in young people. AB - The aim of this study was to develop a reliable self-report instrument to assess obsessive-compulsive disorder (OCD) in young people. The children's Obsessional Compulsive Inventory (CHOCI) had good internal consistency, criterion validity and was significantly correlated with the Children's Yale-Brown Obsessive Compulsive Scale (CY-BOCS). This preliminary new measure could serve to provide a more efficient and accessible way of assessing OCD in young people. PMID- 12551643 TI - Bipseudophakia. PMID- 12551645 TI - Deep lamellar keratoplasty with trypan blue intrastromal staining. PMID- 12551647 TI - Obtaining lenticules for lamellar transplantation. PMID- 12551648 TI - Defining mesopic and scotopic. PMID- 12551650 TI - Postoperative infectious endophthalmitis. PMID- 12551652 TI - Suture fixation of a foldable acrylic IOL. PMID- 12551654 TI - Capsule contraction in polypseudophakia. PMID- 12551655 TI - Consultation section. Refractive surgical problem: left eye LASIK in 25-year old man. PMID- 12551656 TI - Consultation section. Refractive surgical problem: flap folds and DLK. PMID- 12551657 TI - Consultation section. Refractive surgical problem: low corneal thickness to have LASIK. PMID- 12551658 TI - Consultation section. Refractive surgical problem: 3 issues to be discussed: the clinical indications, the surgical procedure, the postoperative findings. PMID- 12551659 TI - Consultation section. Refractive surgical problem: a number of interesting questions. PMID- 12551660 TI - Consultation section. Refractive surgical problem: irregular astigmatism in the left eye. PMID- 12551661 TI - Management of posterior polar cataract. AB - In this technique for managing posterior polar cataract, extreme care is taken not to overpressurize the anterior chamber or capsular bag to prevent posterior capsule rupture. Minimal hydrodissection and hydrodelineation are performed. The nucleus is extracted using minimal ultrasound energy. Viscodissection is used as a primary technique to mobilize the epinucleus and cortex. A protective layer is preserved over the posterior polar region until the conclusion of the extraction procedure to minimize the risk of loss of lens material into the vitreous cavity in the case of a capsule defect. PMID- 12551662 TI - Bacterial endophthalmitis after small-incision cataract surgery. effect of incision placement and intraocular lens type. AB - PURPOSE: To determine whether endophthalmitis after small-incision cataract surgery is affected by the incision site (superior sclerocorneal versus temporal cornea) or the foldable intraocular lens (IOL) material (silicone versus acrylic). SETTING: Multicenter study. METHODS: Patients who had small-incision cataract surgery at Toyama Medical and Pharmaceutical University Hospital and affiliated hospitals from March 1998 to March 2001 were examined prospectively. The patients were randomized into 3 groups. In Group A, an acrylic IOL (MA60BM, Alcon) was implanted through a temporal corneal incision. In Group B, an acrylic IOL (MA60BM) was implanted via a superior sclerocorneal incision. In Group C, a silicone poly(methyl methacrylate) IOL (SI-40NB, Allergan) was implanted via a superior sclerocorneal incision. Each patient was followed for more than 6 months. RESULTS: Initially, 7622 patients (12 317 eyes) agreed to participate in the study. The final study included 3831 eyes in Group A, 3901 eyes in Group B, and 3863 eyes in Group C. Postoperative endophthalmitis was diagnosed clinically in 11 eyes (0.29%) in Group A, 2 (0.05%) in Group B, and 2 (0.05%) in Group C. Endophthalmitis proven by culture was found in 9 eyes in Group A, 2 in Group B, and 2 in Group C. The incidence of endophthalmitis in Group A was higher than in the other groups. The incidence of endophthalmitis in Group B was similar to that in Group C. The relative risk of postoperative endophthalmitis proven by culture in Groups B and C was 4.6 times (P =.037) lower than in Group A. CONCLUSION: The findings suggest that a temporal corneal incision may lead to an increased risk of postoperative endophthalmitis and that the IOL material does not affect the incidence of this complication. PMID- 12551663 TI - Intravitreal triamcinolone acetonide for refractory chronic pseudophakic cystoid macular edema. AB - PURPOSE: To determine the safety and efficacy of intravitreal triamcinolone acetonide (TAAC) injections in patients with refractory cystoid macular edema (CME) after cataract extraction. SETTING: LSU Eye Center, Louisiana State University Health Sciences Center, New Orleans, Louisiana, USA. METHODS: In this nonrandomized retrospective case review, 8 eyes of 8 patients with a history of pseudophakic CME recalcitrant to current standard treatment modalities were enrolled. The mean duration of the CME was 20 months. The patients received intravitreal injections of 1 mg of TAAC and were followed for a mean of 8 months. The main outcome measures included visual acuity, the presence of CME on biomicroscopic examination, angiographic evidence of perifoveal leakage, intraocular pressure (IOP), and complications related to treatment. RESULTS: The visual acuity increased in all patients. The magnitude of improvement was mainly restricted by underlying macular pathology and correlated well with the level of visual acuity at entry into the study. Angiographic improvement occurred in all patients. Temporary increases in IOP were easily controlled with topical medications. No other adverse effects could be attributed to this technique. Repeated injections were required. CONCLUSIONS: Intravitreal administration of TAAC was safe and effective in recalcitrant cases of pseudophakic CME with a beneficial effect on the macular edema and visual acuity. A prospective randomized study is needed to determine with accuracy the efficacy, safety, and exact timing of this technique and possibly to recognize subtypes with a more favorable response. Repeated injections were required in all eyes. The development of a sustained-release intravitreal drug-delivery system would be beneficial. PMID- 12551664 TI - Phacoemulsification of the crystalline lens and implantation of an intraocular lens for the correction of moderate and high myopia: four-year follow-up. AB - PURPOSE: To assess the safety of lens extraction and intraocular lens (IOL) implantation in patients with high myopia treated for initial lens opacity and/or refractive indications. SETTING: Instituto de Microcirugia Ocular de Barcelona, Barcelona, Spain. METHODS: This retrospective nonrandomized case series study comprised 44 eyes of 30 consecutive myopic patients who had surgery because of initial lens opacity and/or refractive indications during a 2-year period. In each case, phacoemulsification was performed using an ultrasonic technique and an IOL was implanted in the capsular bag. The patients were seen preoperatively to evaluate retinal pathology. They also had a complete ophthalmologic evaluation that included detailed indirect ophthalmoscopy. All patients were followed at regular intervals. The main outcome measures were preoperative and postoperative spherical equivalent (SE), the incidence of posterior capsule opacification (PCO) and the need for capsulotomy, and the incidence of retinal complications. RESULTS: In all eyes, the surgery was uneventful. The mean patient age at surgery was 42.83 years; the mean preoperative SE was -15.77 diopters (D) and the mean postoperative SE, -1.05 D. No eye required preoperative peripheral retinal photocoagulation. Twenty-five eyes (56.8%) had PCO and had a neodymium:YAG laser capsulotomy. One eye had a retinal tear 14 months after surgery and was treated with focal photocoagulation. The mean endothelial cell loss was 2.1% during the first postoperative year. Two eyes had an immediate postoperative intraocular pressure (IOP) rise, 1 with an inflammatory membrane and the other with corneal edema; both resolved with topical treatment. One eye with elevated IOP and a bad response to medical treatment had argon laser trabeculoplasty. No eye had a retinal detachment during the follow-up. CONCLUSION: With a thorough preoperative ophthalmologic evaluation and uneventful surgery, patients who have phacoemulsification and IOL implantation for the correction of myopia have a satisfactory chance of obtaining good visual results with few complications. PMID- 12551665 TI - Retinal detachment after cataract extraction and refractive lens exchange in highly myopic patients. AB - PURPOSE: To assess the refractive effects and incidence of postoperative complications in patients with high myopia after cataract extraction. SETTING: University Eye Clinic, Trieste, Italy. METHODS: This noncomparative retrospective study comprised all eyes (N = 388) having cataract extraction and implantation of an intraocular lens with a power less than 11.0 diopters (D) between 1986 and 1999. The preoperative and postoperative refractions; preoperative, postoperative, and final best corrected visual acuity (BCVA); and postoperative complications were analyzed. The mean follow-up was 47.16 months +/- 32.83 (SD) (range 8 to 146 months). RESULTS: Phacoemulsification was performed in 237 eyes, extracapsular cataract extraction in 147 eyes, and intracapsular cataract extraction in 4 eyes. The mean preoperative BCVA was 0.20 +/- 0.21 and the mean postoperative BCVA, 0.58 +/- 0.32. At the final evaluation, the mean BCVA was 0.50 +/- 0.33. The mean refraction was -15.95 +/- 5.86 D preoperatively and -2.00 +/- 1.62 D postoperatively. One eye (0.26%) developed a retinal detachment postoperatively. Of the 127 eyes (32.73%) having a neodymium:YAG laser capsulotomy for posterior capsule opacification, 1 had a macular retinal hole. Posterior detachment of the vitreous occurred in 78.6% of cases. Peripheral retinal photocoagulation was required in 8 cases postoperatively. CONCLUSION: Refractive lens exchange led to good functional results and a low incidence of postoperative complications and can thus be regarded as a viable technique to correct high myopia associated with cataract. However, further study is required before it can be routinely recommended for the correction of high myopia in eyes with a transparent crystalline lens. PMID- 12551666 TI - Outcomes of surgery for posterior polar cataract. AB - PURPOSE: To report the preferred technique and the outcomes of surgery for posterior polar cataract. SETTING: Hayashi Eye Hospital, Fukuoka, Japan. METHODS: Medical records of 28 eyes of 20 consecutive patients with posterior polar cataract who had cataract surgery were reviewed. The surgical techniques used, intraoperative complications, preoperative and postoperative visual acuities, and the causes of impaired visual acuity after surgery were examined. RESULTS: Of the 28 eyes, 25 (89.3%) with a small to medium posterior polar opacity had standard phacoemulsification or aspiration surgery. Two eyes (7.1%) with a large opacity and soft lens nucleus had pars plana lensectomy, and 1 eye (3.6%) with a large opacity and hard nucleus had intracapsular cataract extraction. Among the eyes having phacoemulsification or aspiration surgery, posterior capsule rupture occurred in 2 (7.1%). The mean visual acuity improved significantly after surgery (P <.0001); however, the postoperative visual acuity was worse than 20/20 in 7 eyes (25.0%). The cause of the low acuity was amblyopia in 4 eyes (14.3%), impaired foveal function after retinal detachment in 2 eyes (7.1%), and macular degeneration in 1 eye (3.6%). CONCLUSION: Posterior polar cataracts can be safely extracted if the appropriate surgical technique is selected. Visual acuity improved significantly in most cases, although some patients with a unilateral cataract had previously developed amblyopia. PMID- 12551667 TI - Previous cataract surgery in a defined Swedish population. AB - PURPOSE: To study the prevalence of previous cataract surgery (PCS) in different age groups in a defined Swedish population. SETTING: Department of Ophthalmology, Blekinge Hospital, Karlskrona, Sweden. METHODS: This investigation of the population of Blekinge County, Sweden, was performed in June 2001. All previous cataract extractions in the district were performed at a single surgical unit, and the register of this unit contains data on all consecutive operations since 1980. The population register in the district was studied to ascertain the number of female and male individuals in different age groups for each year from 1980 though 2000. Included were data about deceased patients and those who moved from the area. RESULTS: During 21 calendar years, 10,538 patients had cataract surgery; 6,697 of them (63.6%) were alive in June 2001. The survival rate for patients having cataract surgery during the period shows a linear curve for patients operated on during the past 13 years. Among those in the district aged 70 or older, 23.9% of women and 14.0% of men had cataract extraction in 1 or both eyes. In the population aged 40 years and older, 6.43% had a history of PCS. The annual surgery volume is increasing because of a backlog of cataract cases and because of the yearly incidence. With an increase in the annual surgery volume, it is estimated that given the present indications for surgery, it will take 5 to 10 years to reach a level that means further surgery only in patients having new cataract. CONCLUSION: Even though there is already a much higher prevalence of PCS than is known from the literature, the annual surgery volume continues to increase, even in older age groups. PMID- 12551668 TI - Small-incision manual extracapsular cataract surgery in Ghana, West Africa. AB - PURPOSE: To evaluate the results of small-incision manual extracapsular cataract extraction surgery (ECCE) in a district hospital in West Africa. SETTING: Margret Marquart Catholic Hospital, Ghana, West Africa. METHODS: This prospective study consisted of 200 eyes of 193 patients who had small-incision manual ECCE between January 1999 and May 2000. For comparison, the charts of 32 patients (32 eyes) operated on between July and December 1998 using a limbal incision (control group) were retrospectively analyzed. Outcome measures included intraoperative and postoperative complications, postoperative visual acuity, and refractive astigmatism. RESULTS: In the small-incision ECCE group, self-sealing wounds were achieved in 129 eyes (64.5%). Vitreous loss occurred in approximately 3% of eyes in both the small-incision and control groups. The final visual acuities were similar between the 2 groups, with more than 90% of eyes in both groups achieving a final best corrected visual acuity of at least 20/60. Eyes in the small incision group had faster visual recovery (P <.001), a lower incidence of fibrinous iritis (P =.02), and were more likely to have round pupils (P <.001) than eyes in the control group. The main complication of small-incision surgery was moderate corneal edema, which persisted until at least the 1-week visit in 14 eyes (7%). At the most recent visit, 1 eye in the small-incision group (0.5%) had bullous keratopathy. CONCLUSION: In a district hospital in West Africa, small incision manual ECCE surgery yielded faster visual rehabilitation and had a lower incidence of fibrinous iritis than standard ECCE surgery. PMID- 12551669 TI - Analysis of intraocular lens power calculation in post-radial keratotomy eyes. AB - PURPOSE: To determine whether refractive complications can be prevented by applying the currently most accurate method of intraocular lens (IOL) power calculation in the post-radial keratotomy (RK) patient. SETTING: Department of Ophthalmology, University of California, Davis, Sacramento, and American Eye Institute, Cedars Sinai Medical Center, Los Angeles, California, USA. METHODS: Twenty-four eyes having cataract surgery after RK were studied retrospectively for the final postcataract refraction and for the target refraction used in selecting the IOL. Nine of the eyes were further studied for the keratometry (K) values obtained with different methods and for the theoretical postoperative refraction with an IOL aiming for plano or -1.50 diopters (D) based on the known flatter calculated K, axial length, power of the implanted IOL, and refraction after cataract surgery. RESULTS: Implantation of an IOL aiming for plano in the 24 post-RK eyes would have resulted in a hyperopic refraction in 83.4% cases. The choice of an IOL targeted at myopia reduced the frequency of hyperopia to 42.0% (24 cases). Selection of an IOL calculated with a flatter calculated K and aiming for plano decreased the frequency of hyperopia to 44.4%; however, aiming for 1.50 D still resulted in hyperopia in 44.4% of eyes (9 cases). CONCLUSIONS: Unintentional hyperopia can be significantly decreased but not eliminated as a complication of post-RK cataract surgery. The accuracy of the IOL power determination can be improved if myopia is targeted as the postcataract surgery refractive error and the flatter calculated K is used in the IOL determination. PMID- 12551670 TI - Sex hormones and their receptors in patients with age-related cataract. AB - PURPOSE: To determine the sex hormone levels in the serum and aqueous humor in patients with age-related cataract and the corresponding hormone receptors in cataract lens epithelial cells (LECs). SETTING: Research Laboratory, International Intraocular Implant Training Center, Tianjin Medical University, Tianjin, China. METHODS: Serum and aqueous humor were drawn from patients with cataract and a control group. Enzyme-linked immunosorbent assay was used to determine the levels of estradiol, progesterone, and testosterone in the serum and the aqueous. The anterior lens capsules with attached LECs were obtained during cataract surgery, and the expressions of estrogen receptor, progesterone receptor, or androgen receptor were examined by immunohistochemistry. RESULTS: The testosterone level in the serum was significantly higher in the control group than in the cataract group. In the cataract group, there was no difference between sexes in the serum levels of estradiol or progesterone; however, the testosterone level in men was significantly higher than in women. The aqueous level of each hormone was lower than in serum; however, there was no difference between sexes and no association with corresponding serum levels. No estrogen, progesterone, or androgen receptor was detected in the LECs of patients with age related cataract. CONCLUSIONS: There was no statistical difference between men and women or between the cataract and control groups in the levels of estradiol or progesterone in serum. There was no between-sex difference in the aqueous levels. It appears that sex hormone levels can be regarded only as a risk factor for cataractogenesis, not as a key factor. PMID- 12551671 TI - Thyroid disease and vertical rectus muscle overaction after retrobulbar anesthesia. AB - PURPOSE: To determine the associated factors of persistent diplopia after retrobulbar anesthesia. SETTING: Strabismus Section, Department of Ophthalmology, Seoul Municipal Boramae Hospital, Seoul, Korea. METHODS: A prism and cover test in the diagnostic positions of gaze, force-generation test, and forced-duction test were performed in 11 patients with vertical rectus overaction after retrobulbar anesthesia. A Tensilon (edrophonium chloride) test, thyroid function test, and orbit computed tomography were performed in 8 patients. RESULTS: None of the 11 patients reported diplopia before receiving local anesthesia. Ten patients showed ipsilateral hypertropia with superior rectus overaction and 1 patient, ipsilateral hypotropia with inferior rectus overaction. One patient had partial fibrosis of the medial half of the superior rectus, presumably caused by a bridle suture. Four (50%) of 8 patients in whom a thyroid function test was performed had abnormal findings; 1 had a history of systemic dysthyroidism. CONCLUSIONS: In this study, permanent vertical strabismus after local anesthesia was entirely the result of overactive vertical rectus muscles, mostly the superior rectus muscle. Half the patients who had a thyroid function study had abnormal findings. PMID- 12551672 TI - Optical and ultrasound measurement of axial length and anterior chamber depth for intraocular lens power calculation. AB - PURPOSE: To evaluate the precision, reproducibility, and applicability of an optical method based on partial coherence interferometry for intraocular lens (IOL) power calculation. SETTING: Ultrasound laboratory of a university eye hospital. METHODS: A prospective comparison of measurements made by the IOLMaster optical instrument (Carl Zeiss) and Ultrascan Digital 2000 contact ultrasound A scan (Alcon) for IOL calculations was performed. Examined were 255 eyes of 134 persons (204 phakic, 47 pseudophakic, and 4 aphakic). The mean age of the patients was 67.9 years (range 7 to 94 years). RESULTS: The IOLMaster measurements were successful in more than 80% of cases: in 82%, 99%, and 99% for axial length (AL), anterior chamber depth (ACD), and keratometry measurements, respectively. The reproducibility of the AL and ACD measurements was very high (coefficient of variation 0.13% and 2.20%, respectively). The AL and ACD values were significantly larger with the IOLMaster (P <.001) than with the Ultrascan Digital 2000. The correlation between ultrasound and optical AL measurements was high (r = 0.985; P <.001); however, there was no correlation between ACD measurements (r = 0.079; P =.397). The corneal refractive power measurements of a Javal-type keratometer and the IOLMaster were highly correlated (r = 0.955; P <.001), with a mean difference of 0.2 diopter. CONCLUSIONS: The results show that measurements for IOL calculation are easy and precise with the optical method. It is a noncontact method, so no anesthesia is needed and there is no risk of infection. PMID- 12551673 TI - Comparison of anterior chamber depth measurement methods in phakic and pseudophakic eyes. AB - PURPOSE: To compare anterior chamber depth (ACD) measurements in phakic and pseudophakic eyes using a slit-beam photographic technique (IOLMaster, Carl Zeiss Meditec AG) with those obtained with the laboratory prototype version of partial coherence interferometry (PCI) and with conventional applanation ultrasound in phakic eyes. SETTING: Department of Ophthalmology, Vienna General Hospital, Vienna University, Vienna, Austria. METHODS: Thirty-three ACDs of 28 patients with age-related cataract were measured preoperatively with a slit-beam photographic technique (IOLMaster) and the prototype version of PCI. In 24 eyes, the ACD was also assessed with applanation ultrasound. In addition, 34 ACDs of 18 pseudophakic patients in a different study population were examined postoperatively with the IOLMaster and PCI. RESULTS: The median ACD in the phakic eyes was 3.06 mm (range 1.93 to 3.90 mm) with the IOLMaster, 3.09 mm with PCI (range 1.49 to 4.06 mm), and 2.87 mm (range 2.18 to 3.33 mm) with applanation ultrasound. The precision was 0.005 mm for PCI and 0.015 mm for IOLMaster measurement. The median difference between the IOLMaster and PCI ACD biometry was 0.01 mm +/- 0.14 (SD) (range -0.44 to 0.17 mm) (P =.71). In pseudophakic eyes, the 2 methods showed a median difference of -0.22 mm (range -0.45 to 1.99 mm) (P >.1) and did not correlate (r = 0.21; P >.2). CONCLUSIONS: In phakic eyes, the difference between IOLMaster and PCI measurements was small and not statistically significant. In pseudophakic eyes, the difference was larger and the methods did not correlate. PMID- 12551674 TI - Comparison of 2 A-scans. AB - PURPOSE: To compare 2 A-scan instruments with regard to differences in measured results for the same patient sample. SETTING: St. Erik's Eye Hospital, Stockholm, Sweden. METHODS: In a study to evaluate the lens-haptic plane concept of intraocular lens (IOL) power calculation, 148 patients eligible for cataract surgery were measured with 2 different A-scan instruments (BVI Axis and Sonomed 1500). The axial length (AL), anterior chamber depth (ACD), and lens thickness (LT) results were analyzed for systematic differences and random errors. RESULTS: The Sonomed 1500 measured systematically longer than the BVI Axis for AL (0.41 mm) and ACD (0.28 mm), although the correlation was good (r = 0.99 and r = 0.87, respectively). The LT correlated poorly (r = 0.18) and showed no systematic trend. The relative random errors (standard deviations) in ACD (7.2%) and LT (18.6%) were larger than that of the AL (0.8%). The systematic difference in the AL corresponds to a 1.0 diopter difference in the A-constant. CONCLUSIONS: The large random errors in the ACD and LT reduce their value as predictors of postoperative IOL position in formulas that use them. Systematic differences in AL can be large enough to require separate formula constants for different pieces of equipment. If this is the situation in 1 setting, there is a risk of mistakes. This confusion could be avoided if there were an agreed standard and a universal calibration procedure for instruments intended for AL measurement. PMID- 12551675 TI - Reduction of trend errors in power calculation by linear transformation of measured axial lengths. AB - PURPOSE: To find a method to improve the refractive outcome in short eyes and long eyes without sacrificing the outcome in normal eyes. SETTING: St. Erik's Eye Hospital, Stockholm, Sweden. METHODS: In a prospective study, 148 patients eligible for cataract surgery were measured with 2 different A-scans (BVI Axis, B.V. International; Sonomed 1500, Sonomed Inc.). Refraction was determined 6 weeks postoperatively. The postoperative refraction was compared with the refraction predicted by the Holladay 1, Hoffer Q, and SRK/T formulas; formula constants were optimized to give a zero mean error. The mean absolute error (MAE) was used as an outcome measure. RESULTS: The BVI Axis measured consistently shorter than Sonomed 1500. The mean axial lengths (ALs) were 23.033 mm and 23.435 mm, respectively. With the BVI Axis, an MAE of 0.44 diopter (D), 0.44 D, and 0.47 D was obtained, with the Holladay 1, Hoffer Q, and SRK/T formulas, respectively, with a trend toward undercorrecting short eyes and overcorrecting long eyes. The MAE with the Sonomed 1500 was 0.38 D, 0.39 D, and 0.40 D, respectively. By adding 0.402 mm to each measured value in the BVI Axis data set, the mean AL was transformed to 23.435 mm. With the transformed data, the MAE improved to 0.42 D, 0.43 D, and 0.44 D, respectively, with a reduced trend toward undercorrection and overcorrection. The 0.04 D difference between the instruments, although not statistically significant, may depend on measurement precision. Extending the concept of transformation, a minimum MAE of 0.41 D was obtained with the Holladay 1 at a mean AL of 24.0 mm, 0.43 D with Hoffer Q at 23.9 mm, and 0.40 D with SRK/T at 24.4 mm. The trend toward undercorrection and overcorrection was eliminated at the optimum for each formula. CONCLUSIONS: There were systematic differences in measured AL depending on equipment. Thus, the calculated powers differed and caused error in the degree of compliance between the labeled formula constant of an intraocular lens and the equipment used. Although personalization of formula constants reduces the mean error, in general a trend toward undercorrection of short eyes and overcorrection of long eyes will persist. Transforming the AL scale can eliminate the trend error and improve the overall refractive outcome. Transformation to a population mean AL of about 24.0 mm was close to optimum for the 3 formulas. PMID- 12551677 TI - Comparison of a digital and a handheld infrared pupillometer for determining scotopic pupil diameter. AB - To compare a digital infrared pupillometer with a handheld infrared pupillometer for determining scotopic pupil size. Department of Ophthalmology, Johann Wolfgang Goethe-University, Frankfurt am Main, Germany. In 100 eyes of 50 healthy individuals (mean age 38.8 years +/- 10.7 [SD]), the scotopic pupil size was measured with a digital (Procyon) and a handheld (Colvard) infrared pupillometer. After dark adaptation of 1 minute, measurements were performed with both devices by 2 examiners (E1 and E2). Agreement and repeatability were analyzed using a comparison method described by Bland and Altman. The mean scotopic pupil diameter was 5.90 +/- 0.97 mm (range 3.24 to 7.91 mm) with the Procyon and 5.78 +/- 0.98 mm (range 3.00 to 7.30 mm) with the Colvard pupillometer. The mean difference between the 2 devices was -0.01 mm (E1) and -0.24 mm (E2). The limits of agreement ranged from 2.84 (E1) to 3.24 (E2). The coefficients of repeatability were 0.64 (Procyon) and 1.16 (Colvard). The mean difference between E1 and E2 was -0.10 for the Procyon and 0.13 for the Colvard pupillometer. The limits of agreement ranged from 1.28 (Procyon) to 2.32 (Colvard). The digital infrared pupillometer (Procyon) demonstrated better repeatability and agreement in measuring scotopic pupil size than a handheld device (Colvard). PMID- 12551676 TI - Comparison of 4 methods for quantifying posterior capsule opacification. AB - PURPOSE: To compare the results of posterior capsule opacification (PCO) quantification and the repeatability of a fully automated analysis system (Automated Quantification of After-Cataract [AQUA]) with that of 2 other quantification methods and subjective grading of PCO. A test set of digital retroillumination images of 100 eyes with PCO of varying degrees was used. SETTING: Department of Ophthalmology, University of Vienna, Vienna, Austria. METHODS: One hundred digital retroillumination images of eyes (100 patients) with PCO were selected to attain an even distribution from mild to severe cases. The images were evaluated by 4 methods: subjective grading by 4 experienced and 4 inexperienced examiners, the subjective Evaluation of Posterior Capsular Opacification (EPCO) system, posterior capsule opacification (POCO) software, and the AQUA system. Ten images were presented twice to assess the reproducibility of the analysis systems. RESULTS: Subjective grading correlated best with the subjective EPCO system and the objective AQUA system (r = 0.94 and r = 0.93, respectively). The POCO system showed very early saturation and therefore a much weaker correlation (r = 0.73). The POCO scores reached the maximum of 100% in several minimal to mild PCO cases. The reproducibility of the AQUA software was perfect and that of the other analysis systems, comparably satisfactory. CONCLUSION: The objective AQUA score correlated well with subjective methods including the EPCO system. The POCO system, which assesses PCO area, did not adequately describe PCO intensity and includes a subjective step in the analysis process. The AQUA system could become an important tool for randomized masked trials of PCO inhibition. PMID- 12551678 TI - Objective method to measure corneal clarity before and after laser in situ keratomileusis. AB - PURPOSE: To develop, evaluate, and use an objective method to determine the effect of laser in situ keratomileusis (LASIK) on corneal clarity. SETTING: Centre for Myopia Research, The Hong Kong Polytechnic University, and the Hong Kong Laser Eye Center, Hong Kong, China. METHODS: Color photographs of corneal sections were taken using a digital camera and converted to 8-bit gray-scale images. The desired area of the photograph was isolated using a preset mask, and a gray-scale or corneal clarity index of the desired area was obtained by averaging the "intensity" indices of individual pixels within the area. The reliability of the clarity index measures was determined by comparing test and retest measures. The sensitivity of the method was quantified by its ability to identify a small (clinically undetectable) decrease in corneal clarity produced by tight-fitting soft contact lenses worn for 30 minutes. Finally, corneal clarity was measured and compared in 24 patients before and 1 day, 1 week, and 1 month after LASIK. RESULTS: The reliability value was 4.11 corneal clarity units, and the change in corneal clarity due to soft contact lens use was 16.24 units. In the LASIK patients, there were statistically significant decreases in corneal clarity from preoperatively to 1 day and 1 week but not to 1 month. CONCLUSIONS: The method measured changes in corneal clarity that were undetectable clinically and were 4 times greater than 95% of the differences between test and retest measures. The method is therefore reliable and sensitive. Corneal clarity decreased after LASIK and recovered within approximately 1 month. PMID- 12551679 TI - Central and peripheral corneal thickness measurement with Orbscan II and topographical ultrasound pachymetry. AB - PURPOSE: To compare thickness measurements of the central 6.0 mm of the cornea obtained with the Orbscan(R) II topography system and topographical ultrasound pachymetry. SETTING: School of Optometry, University of Santiago de Compostela, Galicia, Spain. METHODS: In 24 right eyes, pachymetric measurements were taken at the center and 1.2 mm and 3.0 mm on the superior and inferior hemimeridians. A 1 sample t test was applied to assess the significance of the relationship between Orbscan II and ultrasound methods. The relationship between the 2 was assessed by analyzing regression and plotting the differences against the mean corneal thickness. Orbscan II data were analyzed in 3 ways: (1) without the application of an acoustic equivalent correction factor; (2) with a correction factor of 0.92, as recommended by the manufacturer; (3) with correction using the equations derived in this study. The data were systematically compared with those of ultrasound pachymetry. RESULTS: Before the correction factor was applied, the Orbscan II overestimated the corneal thickness at all locations, with the mean difference (48.15 microm +/- 33.74 [SD]) significantly different from zero (P <.001). Differences increased toward the periphery, and the reliability of Orbscan II readings seemed to decrease with thicker corneas. After the acoustic equivalent was applied, the differences were significantly less; however, this effect did not seem clinically significant as large differences remained. When specific corrective equations were applied for each corneal location, the level of agreement between Orbscan II and ultrasound pachymetry improved substantially; the mean (-0.11 +/- 15.22 microm) was not statistically different from zero (P >.05). CONCLUSIONS: The acoustic equivalent correction factor proposed by the manufacturer to obtain corneal thickness measurements with the Orbscan II compared to those from ultrasound pachymetry was not valid for all corneal topography positions. Orbscan II measurements agreed better with those of ultrasound pachymetry when equations for the central and each peripheral location across the topography were applied. PMID- 12551680 TI - Autorefractometry after laser in situ keratomileusis. AB - PURPOSE: To correlate cycloplegic subjective refraction with cycloplegic autorefractometry in eyes that have had laser in situ keratomileusis (LASIK). SETTING: Vlemma Eye Institute, Athens, Greece. METHODS: Subjective refraction and autorefractometry under cycloplegia were performed in 73 eyes of 46 patients 1, 6, and 12 months after LASIK to correct myopia or myopic astigmatism. The preoperative subjective refraction and autorefractometry under cycloplegia in the same eyes served as controls. RESULTS: A statistically significant difference between subjective refraction and autorefraction was found in the sphere and cylinder at all postoperative times. No statistically significant difference was found in the axis. There was no statistically significant difference in the control eyes. CONCLUSIONS: Automated refractometry in eyes that had had LASIK was reliable in the axis only. Retreatments after LASIK should always be based on subjective refraction. PMID- 12551681 TI - Clinical application of the lens opacities classification system III in the performance of phacoemulsification. AB - PURPOSE: To report the correlation of features of cataracts graded by the Lens Opacities Classification System, version III (LOCS III), with phacoemulsification energy expenditure and the balanced salt solution (BSS) volume used during cataract surgery. SETTING: Wolfe Clinic, Marshalltown, Iowa, USA. METHODS: This was a retrospective review of 2364 cases operated on by a single surgeon from January 1998 to July 2000 in which the cataract had been graded at the slitlamp using the 4 grading scales of the LOCS III: nuclear opalescence (NO), nuclear color (NC), cortical cataract (C), and posterior subcapsular cataract (P). Polynomial best-fit lines were derived using regression analysis correlating the 4 preoperative LOCS III characteristics with 3 intraoperative observations: machine-measured phacoemulsification time, mean power expenditure, and BSS volume. RESULTS: As determined by best-fit lines and their coefficient of determination (R(2)), there were exponential relationships between machine measured phacoemulsification time and the degree of NC (R(2) = 0.48) and NO (R(2) = 0.40). Trends existed between NC and NO and the amount of BSS used (R(2) = 0.08 and R(2) = 0.07, respectively). No relationships were observed between the LOCS III classes of cataract, C and P, at any intraoperative observation. CONCLUSIONS: Exponentially, greater phacoemulsification energy was required as NC and NO increased. The LOCS III cataract grading system enhanced the ability to estimate ultrasonic energy expenditure and BSS volume use during phacoemulsification. Preoperative LOCS III cataract classification can help to create a more formally organized, integrated, customized operative plan. PMID- 12551682 TI - Adhesion of soluble fibronectin, vitronectin, and collagen type IV to intraocular lens materials. AB - PURPOSE: To evaluate soluble fibronectin, vitronectin, and collagen type IV adhesion to poly(methyl methacrylate) (PMMA), fluorine-surface-modified PMMA, silicone, hydrophobic and hydrophilic acrylate, and hydrogel intraocular lenses (IOLs) and determine whether hydrophobic and hydrophilic acrylate materials have different fibronectin-adhesion properties. SETTING: Department of Medical Biochemistry, University of Oulu, Oulu, Finland. METHODS: One hundred fifty IOLs were incubated for 1 week at 37 degrees C with radioactive-iodine-labeled soluble fibronectin, vitronectin, or collagen type IV. Fifty IOLs were analyzed for each protein, 5 from each of 10 different IOL models (PMMA, Alcon MC60BM; fluorine surface-modified PMMA, Chiron Fluorilens Centra-55F; silicone, Allergan Medical Optics SI-40NB and Pharmacia and Upjohn CeeOn 911A; hydrophobic soft acrylate, Alcon AcrySof MA60BM and SA30AL and AMO Sensar; hydrophilic soft acrylate, Ioltech Stabibag and Bausch and Lomb BL27; and hydrogel, Bausch and Lomb Hydroview. The amount of adherent protein was measured with a gamma counter at 1 and 7 days and expressed as counts per minute. RESULTS: At 1 week, significantly more fibronectin was bound to the hydrophobic acrylate IOLs than to the 2 hydroxyethyl methacrylate (HEMA) containing hydrophilic acrylate IOLs (P <.05 to.0001). Significantly more vitronectin was bound to the 2 silicone IOLs than to any other IOL (P <.01 to.0001) at 7 days. Collagen type IV adhered best to the hydrophilic acrylate IOLs, which were significantly different (P <.01 to.0001) than the other IOLs at 1 and 7 days. CONCLUSIONS: Each IOL material had a different affinity to each protein. Significant binding to 1 protein does not indicate that the IOL will bind significantly to all proteins; instead, each protein should be studied separately. Fibronectin bound significantly better to hydrophobic acrylate IOLs than to hydrophilic acrylate IOLs, suggesting that the HEMA-containing IOLs should be classified with the hydrogel IOL group. PMID- 12551683 TI - Wound healing in rabbit corneas after photorefractive keratectomy and laser in situ keratomileusis. AB - PURPOSE: To compare the wound-healing process in the rabbit cornea after photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK) with the same refractive correction. SETTING: Department of Ophthalmology, Wakayama Medical University, Wakayama, Japan. METHODS: Adult albino rabbits (N = 24) were used. One eye of each animal had PRK or LASIK with the same refractive correction. Each animal was killed after an interval of up to 6 months. The expression pattern of corneal stromal injury-related molecules with the 2 treatments were compared. Paraffin sections of the cornea were processed immunohistochemically for alpha-smooth muscle actin (alpha-SMA), collagen type IV [alpha1(IV)](2),alpha2(IV), and heat shock protein (HSP) 47 as well as other HSPs. Sections were also examined after hematoxylin and eosin or periodic acid Schiff staining. RESULTS: Hematoxylin and eosin staining showed the central epithelium to be thick in PRK-treated corneas. The thick epithelium was restricted to the area around the corneal flap edge adhesion in LASIK-treated corneas at 3 months. Periodic acid-Schiff staining showed an absence of or interruption in the epithelial basement membrane in PRK-treated corneas for up to 6 months. Heat shock protein 47 was detected in keratocytes on day 3 but not after that in PRK-treated corneas. There was no difference in the expression of other HSPs. Alpha-smooth muscle actin was expressed in keratocytes repopulated in the central anterior cornea of PRK-treated corneas at 28 days. Keratocytes with immunoreactivity for these 2 proteins were not seen in LASIK-treated corneas. Collagen IV [alpha1(IV)](2),alpha2(IV) was not detected in either group of corneas. The central epithelium became transiently thicker in PRK-treated corneas. CONCLUSION: Keratocyte responses to laser stromal ablation were more marked in corneas treated with PRK than in those treated with LASIK. PMID- 12551684 TI - Effect of 3-piece AcrySof and downsized heparin-surface-modified poly(methyl methacrylate) intraocular lenses in infant rabbit eyes. AB - PURPOSE: To evaluate after-cataract formation, ocular growth, and intraocular lens (IOL) behavior in lensectomized infant rabbit eyes implanted with a downsized heparin-surface-modified poly(methyl methacrylate) (HSM PMMA) IOL with long haptics or a 3-piece AcrySof(R) IOL (Alcon Laboratories, Inc.). SETTING: St. Erik's Eye Hospital, Stockholm, Sweden. METHODS: Clear lens extraction was performed in both eyes of 3-week-old rabbits. In Group 1 (n = 9), a downsized HSM PMMA IOL with long haptics was implanted in 1 eye and in Group 2 (n = 9), a 3 piece AcrySof IOL was implanted in 1 eye. The fellow eyes remained aphakic. RESULTS: The amount of after-cataract was significantly less in the eyes with the IOLs than in the aphakic eyes. The eyes with the HSM PMMA IOL had significantly less after-cataract than those with the AcrySof IOL. No significant difference in axial length was noted between the IOL eyes and the aphakic eyes in either group. In the eyes with the 3-piece AcrySof IOL, there was anterior movement of the optic with occlusion of the pupil. CONCLUSIONS: Eyes with the HSM PMMA IOL and the AcrySof IOL had less after-cataract than the aphakic eyes. The HSM PMMA IOL, however, inhibited after-cataract production significantly better than the AcrySof IOL. The 3-piece AcrySof IOL was seriously deformed in the infant rabbit eyes and caused occlusion of the pupil. Surgeons should be cautious in implanting this AcrySof IOL in the eyes of newborns or in eyes with severe microphthalmia. PMID- 12551685 TI - Scanning electron microscopic and histologic evaluation of the AcrySof SA30AL acrylic intraocular lens. Manufacturing quality and morphology in the capsular bag. AB - PURPOSE: To evaluate the properties of the AcrySof(R) SA30AL (Alcon Laboratories, Inc.) single-piece foldable posterior chamber intraocular lens (IOL). SETTING: Center for Research on Ocular Therapeutics and Biodevices, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina, USA. METHODS: Two nonimplanted clinical-quality AcrySof IOLs were examined by gross, light, and scanning electron microscopy (SEM). In addition, 2 eyes implanted with this IOL obtained post-mortem, the first such eyes accessioned in our laboratory and the first described to date, were examined using the Miyake-Apple posterior photographic technique and by histologic sections. RESULTS: Scanning electron microscopy of the SA30AL IOL showed excellent surface finish. The edge of the optic was square (truncated) and had a matte (velvet or ground-glass) appearance, a feature that may minimize edge glare and other visual phenomena. A well fabricated square or truncated optic edge was demonstrated. Miyake-Apple analysis revealed that the SA30AL IOL showed appropriate fit and configuration within the capsular bag. Histologic correlation of the IOL's square edge and its relation to the capsular bag and adjacent Soemmering's ring were noted. CONCLUSIONS: The AcrySof SA30AL IOL is a well-fabricated lens that situates well in the capsular bag. The truncated optic and its relationship to adjacent structures show a morphological profile that has been shown to be highly efficacious in reducing the rate of posterior capsule opacification. PMID- 12551686 TI - Possible adverse effects of drugs used in refractive surgery. AB - Adverse drug reaction reports were compiled from the National Registry of Drug Induced Ocular Side Effects, Casey Eye Institute, Portland, Oregon, USA; the United States Food and Drug Administration, Rockville, Maryland; the World Health Organization, Uppsala, Sweden; and a Medline literature review of the past 50 years. The reported side effects of mydriatics, cycloplegics, benzodiazepines, tetracyclines, iodine, topical anesthetics, medications used for hemostasis, nonsteroidal antiinflammatory drugs, steroids, antibiotics, and artificial tears are described. Physicians should be aware of the side-effect profile and current industry standards for medications used in corneal refractive surgery, including off-label uses. Guidelines are provided for some classes of medications. PMID- 12551687 TI - International intraocular lens and implant registry 2003. AB - This is the fifth annual printing of the International Intraocular Lens and Implant Registry. A total of 1498 intraocular lenses (IOLs) and nonoptical implants from 33 manufacturers are available to surgeons in 2003. PMID- 12551688 TI - Recurrence of keratitis after excimer laser keratectomy. AB - We report 3 patients who experienced a recurrence of nonherpetic keratitis after excimer laser photorefractive surgery. Two patients had a history of culture positive adenoviral keratoconjunctivitis, and 1 had a clinical diagnosis of Thygeson's superficial punctate keratitis (SPK) prior to excimer laser surgery. Patients should be informed that excimer laser surgery may contribute to a recurrence of keratitis and Thygeson's SPK. Recognition and appropriate treatment can result in resolution and maintenance of a good refractive outcome. PMID- 12551689 TI - Conductive keratoplasty to correct residual hyperopia after corneal surgery. AB - Conductive keratoplasty (CK) is an electrical-current-based technique for steepening the central cornea to reduce low to moderate hyperopia. We report 4 patients who had CK to correct hyperopia after laser in situ keratomileusis (LASIK) and were followed for at least 6 months. An overcorrection was noted after the CK procedure in all patients, but no sight-threatening complications arose. Conductive keratoplasty appears to be safe and well tolerated after LASIK. However, the algorithms should be modified to increase the predictability of the CK procedure in previously treated eyes. PMID- 12551690 TI - Localized liquefied after-cataract. AB - We report 2 cases with a new type of after-cataract. In both, a milky white substance accumulated in a semicircular, closed chamber formed by the intraocular lens optic and the posterior lens capsule. Unlike in previous reports, the closed chamber did not cover the entire capsule but was based on residual lens remnants in the equatorial region of the lens capsule. PMID- 12551691 TI - Sudden decrease in vision caused by liquefied after-cataract. AB - We report a case of sudden decrease to unilateral visual acuity in a 39-year-old patient who had extracapsular cataract surgery 5 years earlier. The decrease was caused by involvement of the visual axis because of a rare complication known as liquefied after-cataract. Treatment with a neodymium:YAG (Nd:YAG) laser was successfully performed. Although there was no rupture in the posterior capsule, the opacity disappeared and the visual acuity improved to 20/20. This new form of after-cataract can cause a sudden decrease in vision, and Nd:YAG laser treatment, even without a posterior capsule rupture, can be performed successfully. PMID- 12551692 TI - Recurrence of secondary opacification and development of a dense posterior vitreous membrane in patients with myotonic dystrophy. AB - We present 3 patients diagnosed with myotonic dystrophy who developed recurrences of secondary opacification. One developed a dense posterior vitreous membrane. Patients with myotonic dystrophy may have an increased risk for recurrent secondary opacification and for developing intraocular fibrosis after cataract extraction. PMID- 12551693 TI - Corneal degeneration after silicone oil tamponade in a photorefractive keratectomy treated eye. AB - Three years after uneventful excimer laser photorefractive keratectomy in both eyes, a 34-year-old man sustained a perforating injury in the left eye. The severe injury, followed by endophthalmitis and retinal detachment, necessitated vitrectomy, lensectomy, and an internal tamponade with silicone oil. One month after the accident and operations, disk-shaped corneal edema corresponding to the laser treatment zone appeared. The edematous region showed a circular, ring-like epithelial thickening at the borders and central corneal epithelial erosion. The anterior stroma developed a central opacity that was separated by a clear rim from the developing silicone-oil keratopathy with the typical band shape. PMID- 12551694 TI - Corneal ectasia after LASIK. PMID- 12551695 TI - Traumatic laceration of a posterior capsule. PMID- 12551696 TI - Herpes simplex keratitis after photorefractive keratectomy. PMID- 12551698 TI - Pharmaceutical biotechnology products approved within the European Union. AB - The manufacture of therapeutic proteins represented the first true industrial application of recombinant DNA technology. Thus far some 88 recombinant proteins/monoclonal antibody-based products have gained marketing approval within the European Union (EU). This represents 36% of all new drug approvals since the introduction of the new centralized European drug approval system in 1995. More recently, an increasing proportion of approved proteins are engineered, tailored to display altered pharmacokinetic profiles or reduced immunogenicity in man. Currently no nucleic acid-based products are approved in the EU. Technical innovations/milestones likely characterizing the biopharmaceutical industry within the next decade include approval of some products produced in transgenic systems, approval of some products administered by non-parenteral means, approval of at least some nucleic acid-based products and the identification of novel biopharmaceuticals/biopharmaceutical targets through discoveries in functional genomics and proteomics. PMID- 12551699 TI - The effect of co-administration of adjuvants with a nanoparticle-based genetic vaccine delivery system on the resulting immune responses. AB - Previously, we reported on a novel cationic nanoparticle-based DNA vaccine delivery system. In the present studies, the effects of co-administration of two well-known adjuvants, cholera toxin (CT) and lipid A (LA), with plasmid DNA (pDNA)-coated nanoparticles were investigated. Balb/C mice (n=6) were immunized with either pDNA alone (cytomegalovirus-beta-galactosidase, 5 microg) or pDNA coated nanoparticles with either 0 or 50 microg of LA on days 0, 7, and 14 subcutaneously (s.c.), or topically on shaved skin with either pDNA (5 microg) alone or pDNA-coated nanoparticles with 0, 10, or 100 microg of CT on days 0, 6, 21, and 35. Mice were sacrificed on day 28 or day 45. Serum IgG titer, in vitro cytokine release and cell proliferation of the isolated splenocytes were determined. By the topical route, immunization of mice with 'naked' pDNA together with 10 and 100 microg of CT significantly enhanced the antigen-specific serum IgG titer by four- and 20-fold, respectively, compared to immunization with pDNA alone. Moreover, co-administration of 100 microg CT with the pDNA-nanoparticles enhanced the IgG titer by more than 300-fold over immunization with 'naked' pDNA alone with no CT. In vitro interferon-gamma (IFN)-gamma release from splenocytes isolated from mice immunized with pDNA-coated nanoparticles with CT (100 microg) was increased by three-fold over immunization with pDNA-nanoparticles without CT. Similarly, in vitro IFN-gamma release from splenocytes isolated from mice immunized with 'naked' pDNA with CT (100 microg) was increased by two-fold over immunization with 'naked' pDNA without CT. Finally, pDNA-coated nanoparticles adjuvanted with 10 microg CT resulted in the strongest splenocyte proliferation. By the s.c. route, co-administration of LA (50 microg) with pDNA resulted in more than 16-fold enhancement in IgG titer over immunization with 'naked' pDNA alone. Immunization with pDNA-coated nanoparticles with LA (50 microg) led to 16-fold enhancement in specific serum IgG titer over immunization with pDNA-coated nanoparticles with no LA, and more than 250-fold enhancement over immunization with 'naked' pDNA alone with no LA. Moreover, in vitro IFN-gamma release and proliferation by splenocytes isolated from LA co-immunized mice was also significantly enhanced. In conclusion, CT (topical) and LA (s.c.) are potential adjuvants to further enhance immune responses using a novel cationic nanoparticle based DNA vaccine delivery system. PMID- 12551700 TI - Stability of polycationic complexes of an antisense oligonucleotide in rat small intestine homogenates. AB - Presystemic degradation in the gastrointestinal tract is one of the major problems contributing to the poor oral absorption of antisense oligonucleotides. Complexes between the antisense phosphorothioate oligodeoxynucleotide ISIS 2302 and the polycationic carriers protamine sulfate grade X, protamine chloride grade V, protamine phosphate grade X, poly-L-lysine hydrobromide (PLL), spermidine phosphate salt, spermine diphosphate salt, and Protasan G113 and CL113 were formulated in order to increase stability against intestinal nucleolytic degradation. Specific conductivity measurements were carried out to determine the charge ratio of the complex systems. Nuclease stability assays were performed in a rat small intestine homogenate model, which displayed significant exo- and endonuclease activity. Full-length oligonucleotide and metabolites were analyzed by capillary gel electrophoresis with UV detection at 260 nm. Most of the complexes of ISIS 2302 and the polycationic materials, except PLL-based systems, showed a better protection against enzymatic metabolism than free oligonucleotide. Protamine sulfate and protamine chloride considerably enhanced the nuclease stability of the phosphorothioate antisense oligonucleotide. The association of oligonucleotides with several polycationic substances proved to be an alternative to chemical modification in order to stabilize oligonucleotides in the gastrointestinal tract against nucleolytic degradation. PMID- 12551701 TI - Acylation of peptides by lactic acid solutions. AB - To simplify the search for effective mechanisms to suppress peptide acylation inside drug delivery devices made of poly(D,L-lactic acid) (PLA) and poly(lactic co-glycolic acid), we were looking for a suitable model system that would allow screening of strategies for peptide stabilization. With their low pH and the presence of lactic acid oligomers, diluted lactic acid solutions promised to be a suitable test system that mimics the microclimate in degrading PLA devices. We created solutions of 1-50% (w/w) lactic acid by dilution of concentrated lactic acid. Using high performance liquid chromatography (HPLC) and high performance liquid chromatography coupled with mass spectrometry (HPLC-MS) analysis, oligomer hydrolysis was monitored during the equilibration process of the diluted solutions. Their final oligomer content was determined by titration and by calculations based on HPLC data. HPLC-MS analysis of human atrial natriuretic peptide (ANP) stability in different lactic acid solutions at 37 degrees C for 4 weeks demonstrated that ANP underwent acylation even in diluted solutions containing only 0.05% (w/w) lactic acid oligomers. Purity analysis of lactic acid solutions allowed us to compare the conditions in the solution test-system to the microclimate that prevails inside degrading PLA microspheres. PMID- 12551702 TI - Sustained buccal delivery of the hydrophobic drug denbufylline using physically cross-linked palmitoyl glycol chitosan hydrogels. AB - A physically cross-linked palmitoyl glycol chitosan hydrogel has been evaluated as a controlled release system for the delivery of hydrophobic drugs via the buccal route. Samples of palmitoyl glycol chitosan (GCP) with diminishing hydrophobicity (GCP12>GCP11>GCP21) were synthesized, characterized by (1)H nuclear magnetic resonance and hydrogels prepared by freeze-drying an aqueous dispersion of the polymer in the presence of a model hydrophobic drug denbufylline and in some cases the soluble detergent sodium glycodeoxycholate (GDC). GDC was employed as a penetration enhancer. Gels were analysed for hydration, erosion, mucoadhesion and imaged by scanning electron microscopy. The buccal absorption of denbufylline from GCP12, denbufylline, GDC (20:12:1.5) formulations was also investigated in the rabbit model with Carbopol 974NF (CP), denbufylline, GDC (60:36:4) tablets used as controls. Denbufylline reduced the porosity, erosion and hydration of the gels while GDC increased the hydration and erosion. All gels were mucoadhesive but less so than the control CP tablets. Denbufylline was detected 0.5 h after dosing with the GCP12 formulation and delivery was sustained for at least 5 h after dosing. In comparison delivery from the CP tablets was not sustained and was first detected 1 h after dosing. PMID- 12551703 TI - Oral gingival delivery systems from chitosan blends with hydrophilic polymers. AB - Chitosan blends with hydrophilic polymers including polyvinylalcohol (PVA), polyethyleneoxide (PEO) and polyvinylpyrrolidone (PVP), were investigated as candidates for oral gingival delivery systems. The bioavailabilty conferred by the chitosan blend delivery systems, as concluded from dog studies, was shown to be comparable to that based on chitosan alone, especially for those blends involving high molecular weight hydrophilic polymers. Results from differential scanning calorimetry and dynamic mechanical thermal analysis, Fourier transform infrared spectroscopy and tensile testing, indicated that the chitosan/PEO and chitosan/PVP blends showed evidence of miscibility in all blend ratios studied, while the chitosan/PVA blend only showed evidence of interaction for the (50:50) and (80:20) blends, but not for the (20:80) blend. However, even a phase separated system may show interesting and exploitable properties, as evidenced by the tensile testing data for the high molecular weight PVA blend (20:80). The study also indicated that chitosan blends were superior in other properties compared to chitosan alone. These included improved comfort and reduced irritation, ease of processing, improved film quality, improved flexibility, and enhanced dissolution. Blends of chitosan with different hydrophilic polymers could thus be promising candidates for formulation in oral mucosal delivery systems. PMID- 12551704 TI - pH profiles in human skin: influence of two in vitro test systems for drug delivery testing. AB - Investigations to determine pH profiles across human stratum corneum (SC), in vivo as well as in vitro, were carried out using the tape stripping technique and a flat surface pH electrode. This method was extended to the deeper skin layers (=viable epidermis+dermis; DSL) in vitro. Statistically significant changes in the pH values were detected in the SC between in vivo and in vitro investigations and also between male and female skin in vivo. For the DSL, no gender-dependent differences in pH were observed. While the results achieved for the SC are in accordance with data already published in the literature, the values for the DSL were surprising: An alkaline pH, with a steep increase of about two pH units in the first 100 microm of the DSL and a plateau of this level was thereafter detected. Research was also done to examine the influence of different in vitro test systems on the results of pH measurements across the skin. A permeation model (Franz diffusion cell; FD-C) and a penetration model (Saarbruecken penetration model; SB-M) were compared. Experiments were carried out concerning the incubation time as well as the pH of the acceptor solution in the FD-C. Independent of the test system used, no change in the pH profiles could be observed for the SC, but a strong effect of the acceptor medium and its pH on the pH profiles across the DSL could be demonstrated using the FD-C, which showed itself partly after 30 min in statistically significant differences between incubated and formerly frozen skin. The results after the use of buffer solutions with different pH values, the pH across the DSL seemed to come into line with the one of the buffer solution, which was investigated for acidic as well as alkaline pH values. The results obtained with the flat surface pH electrode were confirmed using two different dyes: the pH-dependent fluorescent dye carboxy-SNARF-1 and the pH indicator bromthymolblue. PMID- 12551705 TI - Comparison of two cyclosporine formulations in healthy Middle Eastern volunteers: bioequivalence of the new Sigmasporin Microoral and Sandimmun Neoral. AB - A study was conducted to establish bioequivalence between two oral cyclosporine 100 mg capsules, Sigmasporin Microoral (Gulf Pharmaceutical Industries - Julphar, United Arab Emirates, under technical co-operation with Sigma Pharma, Brazil) and Sandimmun Neoral (Novartis, Switzerland), in a Middle Eastern population, even though both formulations were found to be bioequivalent earlier in a Brazilian population (data on file). It was designed as a randomized, open label, two-way crossover study in which 30 fasting, healthy male volunteers received a single 100 mg cyclosporine dose with 240 ml of water on two treatment days separated by a 1 week washout period. After dosing, serial blood samples were collected for a period of 24 h. Plasma was analyzed for cyclosporine A by a sensitive, reproducible and accurate HPLC method with MS detection capable of detecting cyclosporine A in the range of 5-400 ng/ml with a limit of quantitation of 5 ng/ml. Various pharmacokinetic parameters including AUC(0-t), AUC(0- proportional, variant ), C(max), T(max), T(1/2), and lambda(Z) were determined from plasma concentrations of both formulations. AUC(0-t), AUC(0- proportional, variant ), and C(max) were tested for bioequivalence after log-transformation of data. No significant difference was found based on ANOVA; 90% confidence intervals (82.98-110.57% for AUC(0-t), 81.57-124.71% for AUC(0- proportional, variant ), 80.15-98.91% for C(max)) for these parameters were found to be within the bioequivalence acceptance range of 80-125%. Based on these statistical inferences, it was concluded that Sigmasporin Microoral is bioequivalent to Sandimmun Neoral. PMID- 12551706 TI - Effects of food on the pharmacokinetics of levodopa in a dual-release formulation. AB - The objective was to assess the effect of food on the pharmacokinetics of levodopa and 3-O-methyldopa after administration of a new levodopa/benserazide formulation with a dual-release drug delivery profile (Madopar DR). In an open label, two-way cross-over study, 19 healthy volunteers who had fasted overnight were randomized to receive a single oral dose of levodopa/benserazide (200/50 mg) in the absence or presence of a standardized, high-fat breakfast, administered 30 min before drug administration. The treatment periods (fasting, non-fasting) were preceded by a baseline regimen of levodopa/benserazide (100/25 mg t.i.d. for 6 or 7 days). Blood samples were taken at specific times over a 12-hour period. Plasma concentrations of levodopa and 3-O-methyldopa were determined by high-performance liquid chromatography for pharmacokinetic evaluation. The parameter C(max) of levodopa was significantly lower and t(max) longer under postprandial conditions than under fasting conditions (mean C(max) 1.41 vs. 2.09 mg l(-1); mean t(max) 3.1 vs. 1.0 h). With food, the area under the curve (AUC) of levodopa was equivalent to that following an overnight fast. Compared with volunteers who had fasted, food did not alter t(1/2). Estimates of C(max), t(max) and AUC of 3-O methyldopa under non-fasting conditions were not significantly different from those under fasting conditions. In conclusion, food decreases the rate of levodopa absorption, but had no effect on the systemic exposure to levodopa and the degree of 3-O-methyldopa formation. Standardization of levodopa/benserazide administration with respect to meal times is recommended. PMID- 12551707 TI - Handling of computational in vitro/in vivo correlation problems by Microsoft Excel II. Distribution functions and moments. AB - MS Excel is a useful tool to handle in vitro/in vivo correlation (IVIVC) distribution functions, with emphasis on the Weibull and the biexponential distribution, which are most useful for the presentation of cumulative profiles, e.g. release in vitro or urinary excretion in vivo, and differential profiles such as the plasma response in vivo. The discussion includes moments (AUC and mean) as summarizing statistics, and data-fitting algorithms for parameter estimation. PMID- 12551708 TI - Influence of beta-cyclodextrin complexation on carbamazepine release from hydroxypropyl methylcellulose matrix tablets. AB - The in vitro release profiles of carbamazepine and beta-cyclodextrin either complexed or simply mixed and subsequently incorporated in hydrophilic matrix tablets containing 15 or 30% hydroxypropyl methylcellulose were evaluated. Solubility studies revealed a linear relationship between the increase in carbamazepine solubility and the increase in beta-cyclodextrin concentration. Drying methods (spray-drying and freeze-drying) were used to obtain carbamazepine/beta-cyclodextrin solid complexes in order to prepare tablets. The results demonstrated that matrix tablets containing carbamazepine/beta cyclodextrin solid complexes displayed faster carbamazepine and beta-cyclodextrin release compared to that containing simple physical mixture. Gelling and matrix formation was impaired in formulation containing 15% hydroxypropyl methylcellulose and spray-dried complex. The comparison of spray-drying and freeze-drying revealed no significant influence of both drying methods on carbamazepine and beta-cyclodextrin dissolution rate when carbamazepine/beta cyclodextrin complexes were incorporated in 30% hydroxypropyl methylcellulose matrix tablets. The results point to the possibility of modulating carbamazepine release using a hydroxypropyl methylcellulose matrix associated to the drug complexed with beta-cyclodextrin. PMID- 12551709 TI - Measurement of pharmaceutical particles using a time-of-flight particle sizer. AB - The Aerosizer is an instrument for time-of-flight measurement, which is widely used in particle size determinations. The results of various studies indicate that there are still some problems related to the optimization of the analysis conditions. In this study, the behaviour of a set of different kinds of pharmaceutical particles during Aerosizer measurements was studied. An Aerosizer LD equipment with an Aero-Disperser was validated with particle size standards. Volume particle size distributions of particles with different size and shape characteristics were determined (PVP, Celphere, lactose, a drug substance, PHB microparticles). The aim was to investigate the effects of the shear force and deagglomeration levels during the dispersion of the particles on the particle size distributions that were obtained. The results of this study indicate that the ability of the instrument to disperse particles is highly dependent on the properties of the materials. According to the validation measurements, the instrument gives accurate results for spherical, uncohesive particles. The capability of the dispersing unit to separate particles aerodynamically was well observed with PVP. Time-of-flight measurements were uncomplicated for relatively large particles, such as Celphere, which have little interaction with each other and with the instrument housing. For lactose, increasing shear force rates resulted in size distributions with larger particle sizes. In the case of the PHB microparticles the results indicated that the aggregates became smaller and particles were partly separated to primary particles with all shear force levels. PMID- 12551710 TI - Photostability of extended-release matrix formulations. AB - The photostability of drugs has been widely studied while less attention is devoted to the possible modifications that UV light may induce on the excipients of a dosage form, in particular, on the functional polymers used to modulate drug delivery. In this work we have evaluated the effect of UV light on the release characteristics of extended-release matrix tablets containing hydroxypropylmethylcellulose (HPMC) or polyethylene oxide (PEO) as retarding polymers. Two different model drugs have been used: nifedipine (insoluble and photolabile drug) and diltiazem (soluble and photostable drug). Photodecomposition of nifedipine was evaluated and the formation of the photoproducts was followed during the dissolution process. Regarding the dissolution stability, the matrix tablets containing HPMC exposed to UV light have not shown significant differences in drug release profiles compared to the same non-irradiated formulation, while the matrix tablets containing PEO and exposed to the same conditions of UV light have shown a remarkable increase of drug release rate within the first minutes of the dissolution test (burst effect) which is particularly critical because it can cause the loss of the desired therapeutic control. PMID- 12551711 TI - AFM imaging of calixarene based solid lipid nanoparticles in gel matrices. AB - Contact mode atomic force microscopy has been carried out on gels of four current polymers, carbopol 980, carbopol 2020, hyaluronic acid and xanthan containing dispersions of solid lipid nanoparticles (SLNs) of amphiphilic calixarenes. Imaging shows that the SLNs are dispersed within the gels as discrete particles of 150nm in diameter and show little or no aggregation. The simultaneous use of lateral force, topographic and force modulation mode imaging allows a clear interpretation of the observed images, showing the presence of nanoparticles in the sub-surface region and that the SLNs affect the local mechanical properties of the gels. Photon correlation spectroscopy similarly demonstrates a lack of interaction in suspensions. PMID- 12551712 TI - Preparation and characterization of sterile sub-200 nm meso-tetra(4 hydroxylphenyl)porphyrin-loaded nanoparticles for photodynamic therapy. AB - A photosensitizer, meso-tetra(4-hydroxyphenyl)porphyrin, was incorporated into sub-150 nm nanoparticles using the emulsification-diffusion technique in order to perform sterilization by filtration using 0.22 microm membranes. The three selected polyesters (poly(D,L-lactide-co-glycolide), (50:50 PLGA, 75:25 PLGA) and poly(D,L-lactide (PLA)) for the nanoparticle production were all amorphous in nature and have similar molecular weights but different copolymer molar ratios. The influence of the copolymer molar ratio and the theoretical drug loading was investigated in terms of particle size, drug loading, entrapment efficiency and surface characteristics. With all the polymers used, sub-150 nm nanoparticles were produced with good reproducibility and narrow size distributions irrespective of both the polymer nature and the theoretical drug loading. After purification by cross-flow filtration, the nanoparticle suspensions were sterilized by membrane filtration and freeze-dried in the presence of a lyoprotectant (trehalose). For all types of nanoparticles, complete redispersion in various media could be obtained. All final freeze-dried products were refiltrable on a 0.22 microm membrane and were stable in terms of mean particle size and drug loading over a period up to 6 months. The effective drug loading increased at higher theoretical drug loading, the entrapment efficiency was however decreased. The same trend was observed with the three polyesters. The sterility of the final freeze-dried nanoparticles was confirmed by the results of the sterility testing which showed no bacterial contamination. PMID- 12551713 TI - Solvent injection as a new approach for manufacturing lipid nanoparticles- evaluation of the method and process parameters. AB - Lipid nanoparticles (LNP) can be prepared by rapidly injecting a solution of solid lipids in water-miscible solvents or a water-miscible solvent mixture into water. The aim of the present study was to evaluate the potential of this method for the preparation of LNP and the physicochemical characterization of the particles produced by this method. The results show that solvent injection is a potent and versatile approach for LNP preparation. Acetone, ethanol, isopropanol and methanol are suitable solvents in contrast to ethylacetate with which no LNP could be prepared. The obtained particle sizes (z-average) were between 80 and 300 nm depending on the preparation conditions. Up to 96.5% of the employed lipid was directly transformed into LNP. The LNP formation process seems to be diffusion controlled. Physicochemical characterization of the particles by differential scanning calorimetry (DSC), transmission electron microscopy and X ray diffraction analysis reveals a distinct decrease in crystallinity of the colloidal lipid in comparison to the bulk lipid. Furthermore, DSC analysis of LNP hints at a delayed recrystallization of the colloidal lipid and the presence of two modifications. Therefore, a certain physical instability of the LNP has to be considered. PMID- 12551714 TI - Monolithic glyceryl trimyristate matrices for parenteral drug release applications. AB - Monolithic lipid matrices were developed that allow parenteral drug release for days, weeks or even months. The cylindrical matrices consist of triglycerides or triglyceride/cholesterol mixtures and allow, due to their small dimensions, an application via injection. Pure triglyceride matrices showed less than 3%, triglyceride matrices containing 70% and more cholesterol less than 10% water uptake over 30 weeks. This swelling behavior would allow the use of such matrices even for sophisticated applications such as interstitial drug delivery to the brain where excessive swelling is highly undesirable. The drug release kinetics were found to depend strongly on the fatty acid chain length of the triglyceride and the cholesterol content of the matrices. Increasing the chain length from C(12) to C(18) allowed an increase in the release of pyranine, a low molecular weight model compound, from approx. 60 days to more than 120 days. Adding cholesterol to glyceryl trimyristate matrices made it possible to adjust the release within a time span varying from days to weeks. While matrices containing 50% cholesterol released pyranine within 8 days, cholesterol contents of 90% allowed a release of the dye for more than 3 weeks. PMID- 12551719 TI - Biphasic changes in the Ser-9 phosphorylation of glycogen synthase kinase-3beta after electroconvulsive shock in the rat brain. AB - BACKGROUND: Glycogen synthase kinase-3beta (GSK-3beta) plays important roles in intracellular signaling pathways. Phosphorylation at Ser-9 reduces the activity of GSK-3beta, while phosphorylation at Tyr-216 enhances its activity. Mood stabilizing agents increase the phosphorylation of GSK-3beta at Ser-9, and hence inhibit its activity. This property has been considered to be related to the therapeutic action of these drugs. The effect of electroconvulsive shock (ECS), an effective mood stabilizing treatment, on the phosphorylation of GSK-3beta is not known yet. METHODS: In this study, the effect of ECS on the phosphorylation of GSK-3beta was examined in the rat frontal cortex, hippocampus, and cerebellum by Western blot analysis using antibodies specific for Ser-9 or Tyr-216 phosphorylated GSK-3beta. RESULTS: In all regions, the phosphorylation of GSK 3beta at Ser-9 was decreased immediately after ECS, but then increased above basal level within 10 min and maintained at an increased level for more than 30 min. Meanwhile, the phosphorylation at Tyr-216 of GSK-3beta did not show any significant changes after ECS. CONCLUSIONS: These results showed that ECS could induce biphasic changes in the Ser-9 phosphorylation of GSK-3beta in the rat brain, suggesting some similarities, as well as differences, in the actions of mood stabilizers and ECS in the signal transduction mechanisms of the brain. PMID- 12551720 TI - Strain-dependent modification of behavior following antidepressant treatment. AB - The effects of repeated antidepressant drug treatment on behavioral outcome in Wistar Kyoto (WKY) rats, a putative animal model of depressive behavior, were compared to Wistar and Sprague-Dawley (SD) rats. Rats were treated with desipramine (norepinephrine [NE] uptake blocker), nomifensine (NE and dopamine [DA] uptake blocker), paroxetine (serotonin [5-HT] uptake blocker) or saline, for 12 days. On Day 11, rats were tested in the Porsolt forced swim test (FST). On Day 12, rats were tested in the open field test (OFT). Stress reactivity was assessed on Day 13 when all rats were exposed to water-restraint ulcerogenic stress. Significant strain differences in behavioral responses to the drug treatments were observed. Control WKY rats showed the typical freezing behavior in the OFT and excessive floating behavior in the FST as compared to Wistar and SD rats. Desipramine and nomifensine decreased immobility and increased swim time in the FST in WKY rats. Nomifensine reduced response latency in the OFT in WKY rats and increased activity in the OFT in WKY and SD rats. None of the drugs altered the FST in SD rats. Following ulcerogenic stress, desipramine was the only antidepressant that decreased ulcer incidence in all rat strains compared to saline controls. These results suggest that the "depressive behavior" in WKY rats may be modified by antidepressants that alter synaptic levels of NE and/or DA, but not 5-HT. PMID- 12551721 TI - Effects of electroconvulsive therapy on regional cerebral blood flow measured by 99mtechnetium HMPAO SPECT. AB - Decreased cerebral blood flow is documented in depression. In this study, 15 patients were studied with 99mTc-HMPAO SPECT before and after a course of electroconvulsive therapy (ECT). Improvement in frontal and temporal hypoperfusion was seen only in those patients who responded to ECT. Cerebral hypoperfusion may be a state-dependent marker in depressive illness. PMID- 12551722 TI - Gender-specific effects of social housing in rats after chronic mild stress exposure. AB - Stress plays an important role in the development of affective disorders. Women show a higher prevalence for these disorders than men. The course of a depression is thought to be positively influenced by social support. The authors have used a chronic mild stress model in which rats received footshocks daily for 3 weeks. Since rats are social animals we hypothesized that "social support" might reduce the adverse effects of chronic stress. To test this hypothesis, male and female rats were housed individually or socially in unisex groups of four rats. An open field test was repeated four times during the 3 weeks of treatment. Neuronal activation in the paraventricular nucleus of the hypothalamus (PVN) and dorsal raphe nucleus (DRN) in response to stress was measured the last day with c-fos. Chronic stress exposure increased locomotor activity in the open field, especially during the first minute. This was most pronounced in the individually housed females. In females, social housing prevented the stress-induced increase of locomotor activity, while in males social housing had no effect. Fos immunoreactive (FOS-ir) in the PVN was increased in all stress-exposed groups, except for the socially housed females due to a higher FOS-ir in controls. Individually housed males and socially housed females showed increased FOS-ir in the DRN and the increase was almost significant in socially housed males. IN CONCLUSION: These results show that social housing can enhance coping with stress in female rats, whereas in male rats group housing did not have a positive influence on stress-sensitivity. PMID- 12551723 TI - Reduced basal and phencyclidine-induced expression of heat shock protein-70 in rat prefrontal cortex by the atypical antipsychotic abaperidone. AB - The effect of antipsychotic treatment on basal and phencyclidine (PCP)-induced heat shock protein-70 (hsp70) mRNA expression was studied in the rat striatum and in the prefrontal cortex. Abaperidone, a novel drug with an atypical antipsychotic profile, was compared, at pharmacologically equivalent doses, with the atypical antipsychotics clozapine and risperidone and also with haloperidol, a classical antipsychotic. Abaperidone and clozapine reduced basal hsp70 mRNA expression in the rat striatum and in the prefrontal cortex. No change in either region was found after haloperidol, whereas risperidone reduced hsp70 mRNA in the striatum but not in the prefrontal cortex. The N-methyl-D-aspartate (NMDA) receptor antagonist PCP significantly elevated hsp70 mRNA levels in the prefrontal cortex, an elevation that was potentiated by haloperidol and prevented by all of the atypical antipsychotics tested. Since hsp70 has been associated to some schizophrenia symptoms, we suggest that reduced hsp70 in the prefrontal cortex, a cortical area that plays a critical role in the etiology of many schizophrenia symptoms, may be linked to an atypical profile of antipsychotics, such as clozapine, and possibly also abaperidone. PMID- 12551724 TI - Audioverbal cognitive dysfunction in depression. Factors involved. AB - There is increasing evidence suggesting that depression is associated with a certain degree of cognitive dysfunction. However, there is still debate on whether this dysfunction is only substantially associated with the most severe forms of depression, on whether or not it decreases in parallel with clinical response, and on the role played in these changes by psychotropic medications. In order to clarify these questions, we analyzed the performance in several cognitive tasks that involved attention and working memory of 40 untreated subjects with a diagnosis of dysthymia or major depressive disorder without melancholia. The protocol used included three audioverbal tasks: vocal reaction time (VRT), inverse spelling (IS) and text repetition (TR). The protocol was also administered to 20 healthy volunteers that were used as a comparison group. The same battery of assessments was administered 2 months later to all 60 subjects. At the time of the second assessment, patients (but not healthy volunteers) were on antidepressant medication, in accordance with common clinical practice. The authors found a longer VRT in patients versus healthy volunteers at baseline. VRT did not decrease in patients that responded to treatment. However, there was an improvement in VRT in patients that took sertraline (n=16) compared to subjects taking imipramine (n=11). This fact was not attributable to differences in antidepressant response. Performance in the two other tasks was globally worse in the patient group than in the comparison group, and there was also an absence of improvement in the scores of patients who responded to treatment. However, when the sample was stratified by illness duration, individuals with less than 10 years from the first episode of depression showed a decrease in IS errors compared to the healthy volunteers. It is concluded that patients with nonmelancholic depression suffer from cognitive dysfunction, that this dysfunction persists after clinical improvement and that at least attention is influenced by the type of medication taken. Time from onset of the disorder also seems to influence changes in cognitive performance. PMID- 12551725 TI - Treatment with the atypical antipsychotic, olanzapine, prevents the expression of amphetamine-induced place conditioning in the rat. AB - Place conditioning (PC) experiments were conducted as a means to further elaborate the treatment potential of the atypical antipsychotic, olanzapine (OLZ), for stimulant abuse. The resulting preference/aversion provides an indirect measure of the incentive salience (i.e., euphoria/dysphoria) produced by a drug. Male Sprague-Dawley rats (n=48) were conditioned in two unique environments (i.e., vertical vs. horizontal stripped walls, large vs. small grid flooring) using injections (1.0 mg/kg ip) of either amphetamine (AMPH) or saline (SAL). On average, animals displayed a significant preference for the AMPH-paired location after 2.5 weeks of conditioning (five pairings each of AMPH and SAL). Once the preference was established, animals were pretreated (60 min) with a single dose of OLZ (0.0, 0.56, 1.0 or 1.5 mg/kg sc) given on the test (AMPH-free) day. For the following week's test, animals were injected with SAL (1.0 mg/kg ip) in an attempt to recapture the side preference exhibited before OLZ treatment. OLZ treatment prevented the expression of the AMPH-conditioned preference and reduced locomotor activity. Inhibition of preference resulted from the highest dose of OLZ (1.5 mg/kg), while the inhibition of locomotor activity occurred across all three doses. Additionally, while the effects on preference were no longer apparent by the SAL test the following week (reversible), the activity was still depressed during the SAL tests in animals that had experienced the highest dose of OLZ (1.5 mg/kg). Control experiments, in which OLZ was used as the conditioning drug, suggest that OLZ itself possesses no aversive effects in the PC paradigm, and may even produce a preference for the drug-paired chamber. Because the AMPH preference is dependent on dopamine (DA) release in the nucleus accumbens (NAcc), these experiments suggest that OLZ pretreatment interferes with the rewarding, as well as the subjective effects of AMPH. PMID- 12551726 TI - Regional changes in rat brain inositol monophosphatase 1 (IMPase 1) activity with chronic lithium treatment. AB - Myo-inositol monophosphatase 1 (IMPase 1) is one of the targets for the mood stabilizing action of lithium. Inhibition of IMPase is the basis for the "inositol depletion hypothesis" for the molecular action of lithium. To better understand the precise action of chronic (up to 4 weeks) lithium treatment on IMPase 1 activity, we measured IMPase 1 activity using both a colorimetric and a radiometric assay in rats (53-58 days old) fed a diet containing 0.2% lithium carbonate. Our results show that IMPase 1 activity increases substantially in the various brain regions analyzed, even doubling in some regions in the following order, after chronic treatment: hippocampus>cerebellum>striatum>cerebral cortex>brain stem. Both the qualitative and quantitative increases of IMPase 1 activity by chronic lithium treatment were substantiated by Western blot analysis of hippocampal and cerebral cortex regions. We conclude that the increased IMPase 1 activity is an adaptational response to chronic lithium treatment, and may involve direct or indirect stimulation of IMPA1 (which encodes IMPase 1) and/or turnover of the enzyme. The increased enzyme activity may alter critical neurochemical processes involving either free myo-inositol, the precursor of inositol based signaling system or other metabolic pathways, since IMPase 1 also utilizes selective sugar phosphates, such as galactose-1-phosphate, as substrates. One or more of these signal and metabolic pathways may be associated with lithium's psychotherapeutic mood-stabilizing action. PMID- 12551727 TI - Acetylcholine reduces cyanide-induced superoxide anion generation and lipid peroxidation in rat brain homogenates. AB - Alzheimer's disease (AD) is known to be characterised by a decrease in several brain neurotransmitters. However, the neurotransmitter, which decreases most markedly in this disorder, is acetylcholine (Ach). In addition, the biosynthetic enzyme for Ach, viz. choline acetyltransferase, is also decreased significantly. The ultimate loss of cholinergic neurons and subsequent neurodegeneration in this disorder is thought to be due to free radicals. Presently, there are no reports on the neuroprotective effects of Ach. If Ach has neuroprotective effects, its decline could leave the brain exposed to insults such as free radical damage. Thus, in the present study, the authors determined the ability of Ach to scavenge free radicals using the nitroblue tetrazolium (NBT) assay and also examined its ability to inhibit KCN-induced lipid peroxidation in rat brain homogenates. The results show that Ach inhibits the KCN-induced rise in free radicals and lipid peroxidation. Thus, Ach appears to have neuroprotective properties and this could thus be one reason for the acceleration in neurodegeneration once the levels of this neurotransmitter decline in AD. PMID- 12551728 TI - Mechanisms of cellular distribution of psychotropic drugs. Significance for drug action and interactions. AB - Distribution of a drug in the body is dependent on its permeation properties, the blood flow rates in various tissues, and on plasma and tissue uptake. The distribution of drugs in vivo is largely determined by uptake competitions between blood and tissues, as well as competitions among individual tissues. Basic lipophilic drugs are characterized by extensive accumulation in tissues, which leads to a high volume of distribution. Nonspecific binding to cellular membranes and uptake by acidic compartments (mainly lysosomes) are responsible for such a distribution pattern. Lysosomal trapping is an important mechanism of distribution of basic psychotropic drugs; however, the tissue distribution of the aliphatic-type phenothiazine neuroleptic promazine, tricyclic antidepressants (TADs) and selective serotonin reuptake inhibitors (SSRIs) depends more on phospholipid binding than on lysosomal trapping, whereas in the case thioridazine and perazine, lysosomal trapping is as important for the tissue uptake as is phospholipid binding. Neuroleptics and antidepressants mutually inhibit their lysosomal uptake. A decrease in the intralysosomal drug concentrations in vivo leads to a shift of the drug from organs abundant in lysosomes (lungs, liver and kidneys) to those poor in these organella, e.g., the heart, which may be of clinical importance (cardiotoxicity). The brain is not a homogenous organ, i.e., the phospholipid pattern and density of lysosomes vary in its different regions. Therefore, the contribution of the two mechanisms (lysosomal trapping and tissue binding) to total drug uptake is different in areas of the brain. Both lysosomal trapping and binding to cellular elements for psychotropics are higher in the grey matter and neurons than in the white matter and astrocytes, respectively. Lysosomal trapping and distribution interactions of psychotropics take place mainly in neurons. A decrease (via a distributive interaction) in the concentration of psychotropics in lysosomes (depot) may lead to an increase in their level in membranes and tissue fluids (i.e., in concentrations and compartments relevant to their pharmacological action) and, in consequence, to enhancement of the drug binding to neurotransmitter receptors and/or transporters. PMID- 12551729 TI - Recurrent brief depression--past and future. AB - Recurrent brief depressive disorder (RBD) is a well-defined and significantly prevalent affective disorder with an increased risk of suicidal behavior and significant clinical impairment in the community and general practice. RBD is characterized by depressive episodes occurring at least once a month and lasting for only a few days. The lifetime co-occurrence of both RBD and major depressive disorder (MDD), called combined depression (CD), increases substantially the risk for suicide attempts, even more than is known for "pure" MDD. Diagnostic criteria for RBD can be found in the ICD-10 and DSM-IV and are helpful in both, research and clinical routine. Furthermore, several methodological issues are covered in this paper, which make clinical diagnostic and drug response evaluation of RBD very different from MDD. However, clinical procedures rather bear a resemblance to those used in the treatment of migraine or epilepsy. Formal differences in the course of RBD and MDD create different needs concerning the design of drug treatment studies. Absence of special methodological requirements and highly selected patient samples has probably been responsible for false negative results in double-blind, placebo-controlled treatment studies. Although several authors reported successful treatment of RBD with different compounds in about 60 patients, it is still not possible to deduce a treatment algorithm for RBD to date. Obviously, future treatment studies without the limitations of previous studies are clearly required for RBD. PMID- 12551730 TI - Role of selective serotonin reuptake inhibitors in psychiatric disorders: a comprehensive review. AB - The selective serotonin reuptake inhibitors (SSRIs) have emerged as a major therapeutic advance in psychopharmacology. As a result, the discovery of these agents marks a milestone in neuropsychopharmacology and rational drug design, and has launched a new era in psychotropic drug development. Prior to the SSRIs, all psychotropic medications were the result of chance observation. In an attempt to develop a SSRI, researchers discovered a number of nontricyclic agents with amine uptake inhibitory properties, acting on both noradrenergic and serotonergic neurons with considerable differences in potency. A given drug may affect one or more sites over its clinically relevant dosing range and may produce multiple and different clinical effects. The enhanced safety profile includes a reduced likelihood of pharmacodynamically mediated adverse drug-drug interactions by avoiding affects on sites that are not essential to the intended outcome. SSRIs were developed for inhibition of the neuronal uptake pump for serotonin (5-HT), a property shared with the TCAs, but without affecting the other various neuroreceptors or fast sodium channels. The therapeutic mechanism of action of SSRIs involves alteration in the 5-HT system. The plethora of biological substrates, receptors and pathways for 5-HT are candidates to mediate not only the therapeutic actions of SSRIs, but also their side effects. A hypothesis to explain these immediate side effects is that 5-HT is increased at specific 5-HT receptor subtypes in discrete regions of the body where the relevant physiologic processes are regulated. Marked differences exist between the SSRIs with regard to effects on specific cytochrome P450 (CYP) enzymes, and thus the likelihood of clinically important pharmacokinetic drug-drug interactions. Although no clear relationship exists between the clinical efficacy, plasma concentration of SSRIs, nor any threshold that defines toxic concentrations, but therapeutic drug monitoring (TDM) may be useful in special populations, such as in elderly patients, poor metabolizers (PM) of sparteine (CYP2D6) or mephenytoin (CYP2C19), and patients with liver and kidney impairment. Several meta-analyses have reviewed the comparative efficacy of TCAs and SSRIs, and concluded that both TCAs and SSRIs have similar efficacy in the treatment of depression. SSRIs have demonstrated better efficacy and tolerability in the treatment of obsessive compulsive disorder (OCD). They have also been found to be effective in the treatment for social anxiety disorder both in reducing total levels of social anxiety and in improving overall clinical condition. The benefit of SSRIs in anorexia nervosa (AN) is apparently short-term unless medication is given in the context of nutritional or behavioral therapy. No single antidepressant can ever be recommended for every patient, but in a vast majority of patients, SSRIs should be considered as one of the first-line drugs in the treatment of depression. PMID- 12551731 TI - Alpha-synuclein, Abeta and Alzheimer's disease. AB - Alpha-synuclein is a presynaptic protein that is implicated in the pathogenesis of various neurodegenerative diseases. Missense mutations in the alpha-synuclein gene are linked to familial cases of Parkinson's disease (PD), and it has further been shown that alpha-synuclein is a major constituent of the Lewy bodies in sporadic PD and dementia with Lewy body (DLB). The contribution of alpha synuclein to the pathological changes in Alzheimer's disease (AD) has been currently a matter of scientific debate. Some reports hypothesized that alpha synuclein may play a role in amyloid beta/A4 protein (Abeta) aggregation in senile plaques, whereas recent reports challenged this finding by showing a lack of alpha-synuclein-immunoreactivity in Abeta plaques. In this review, we report on recent findings on the physiological and pathological role of alpha-synuclein and try to elucidate its possible contribution to AD pathology. PMID- 12551732 TI - Vitamin E attenuates reserpine-induced oral dyskinesia and striatal oxidized glutathione/reduced glutathione ratio (GSSG/GSH) enhancement in rats. AB - The effects of a previous and concomitant treatment with vitamin E (VE) were studied on an animal model of tardive dyskinesia, i.e., the frequency of spontaneous tongue protrusions in rats treated with reserpine (RE). VE (5, 10, 20 or 40 mg/kg administered intraperitoneally, daily, for 19 days) attenuated the increase in tongue protrusion frequency induced by RE (1 mg/kg administered subcutaneously, on Days 16 and 18, 1 h after VE), which was quantified on Day 19. In a second experiment, a similar treatment with 20 mg/kg VE attenuated RE induced increase in the striatal ratio of oxidized/reduced glutathione (GSSG/GSH), an index of the oxidative stress process. These results support the free radical hypothesis of tardive dyskinesia. PMID- 12551733 TI - Valproate-associated reproductive and metabolic abnormalities: are epileptic women at greater risk than bipolar women? AB - OBJECTIVE: Evidence indicates that valproate (VPA) may have an adverse impact on reproductive endocrine and metabolic functions in women with epilepsy. This study explores whether the association of VPA with reproductive endocrine abnormalities is applicable to women with bipolar disorder (BD) or is unique to women with epilepsy. METHODS: Thirty female patients aged 18-40 years with a DSM-IV diagnosis of BD (15 on lithium monotherapy and 15 on VPA monotherapy or VPA in combination with lithium therapy) and 15 with idiopathic generalized epilepsy (IGE) on VPA monotherapy were evaluated for reproductive endocrine functioning and metabolic parameters. RESULTS: The menarche age, mean length of menstrual cycle and mean length of menses were not significantly different between groups. None of the bipolar patients on lithium, three (20%) of the bipolar patients on VPA and seven (47%) of the epileptic patients on VPA reported menstrual disturbances. Hirsutism scores of the epilepsy group were significantly higher than those bipolar women, regardless of treatment. Serum total testosterone levels were significantly higher in patients (both with BD and with IGE) treated with VPA than in those treated with lithium. Serum FSH levels were significantly lower and LH-to-FSH ratio was significantly higher in patients with epilepsy than in patients with BD, regardless of treatment. The weight parameters and lipid values investigated did not differ significantly between the groups. CONCLUSION: The study supports the conclusion that VPA may be associated with menstrual abnormalities and increased total testosterone levels in both bipolar and epileptic patients although women with BD did not show clinical features of hyperandrogenism (menstrual abnormalities, hirsutism and truncal obesity) as did frequently as women with epilepsy. PMID- 12551734 TI - Comparison of Lavandula angustifolia Mill. tincture and imipramine in the treatment of mild to moderate depression: a double-blind, randomized trial. AB - Depression is a major health problem and is not only underrecognized and undertreated but is associated with significant morbidity and mortality. Lavandula angustifolia Mill. (Lamiacae) is used to treat depression. Many medicinal plant textbooks refer to this indication, whereas there is no evidence based document. Our objective was to compare the efficacy of tincture of L. angustifolia with imipramine in the treatment of mild to moderate depression and to evaluate the possible adjuvant effect of this tincture in a 4 week double blind, randomized trial. Forty-five adult outpatients who met the Diagnostic and Statistical Manual of Mental Disorders, 4th edition, for major depression based on the structured clinical interview for DSM IV participated in the trial. Patients have a baseline Hamilton Rating Scale for Depression score of at least 18. In this double-blind, single-center trial, patients were randomly assigned to receive lavandula tincture (1:5 in 50% alcohol ) 60 drops/day plus placebo tablet (Group A), tablet imipramine 100 mg/day plus placebo drop (Group B) and tablet imipramine 100 mg/day plus lavandula tincture 60 drops/day (Group C) for a 4-week study. In this small preliminary double-blind and randomized trial, lavandula tincture at this concentration was found to be less effective than imipramine in the treatment of mild to moderate depression (F=13.16, df=1, P=.001). In the imipramine group, anticholinergic effects such as dry mouth and urinary retention were observed more often that was predictable, whereas headache was observed more in the lavandula tincture group. A combination of imipramine and lavandula tincture was more effective than imipramine alone (F=20.83, df=1, P<.0001). As this study indicates, one of the advantages of this combination is a better and earlier improvement. The main overall finding from this study is that lavandula tincture may be of therapeutic benefit in the management of mild to moderate depression as adjuvant therapy. A large-scale trial is justified. PMID- 12551735 TI - Depressive mixed state: dimensional versus categorical definitions. AB - BACKGROUND: Recently, there has been a rebirth of studies on depressive mixed state (DMX), defined as a major depressive episode (MDE) plus few concurrent hypomanic symptoms. It is still unclear how to best define DMX. The study's aim was to test a categorical versus a dimensional definition of DMX. METHODS: Consecutive 260 bipolar II disorder and 173 unipolar MDE outpatients were interviewed with the Structured Clinical Interview for DSM-IV, when presenting for MDE treatment (drug-free). Hypomanic symptoms during index MDE were systematically assessed and graded by a hypomania rating scale (Hypomania Interview Guide, HIG). Different cutoffs of the HIG to define DMX were tested versus a categorical definition of DMX (requiring more than two concurrent hypomanic symptoms, DMX3). Sensitivity and specificity for predicting bipolar II diagnosis were compared. The best definition of DMX based on the HIG was also compared to DMX3 versus typical bipolar variables (early onset, many recurrences, atypical features, bipolar family history). RESULTS: An HIG cutoff of 8 had a specificity for predicting bipolar II diagnosis similar to that of DMX3. HIG>8 was strongly associated with bipolar family history (an important external diagnostic validator), and DMX3 was significantly associated with more bipolar variables (including bipolar family history). LIMITATION: The interview was done by a single interviewer. CONCLUSIONS: Similar specificity for predicting bipolar II disorder diagnosis and a similar strong association with bipolar family history suggest that a categorical and a dimensional definition of DMX could have similar validity. However, the dimensional definition (based on the scoring of a hypomania rating scale) could lead to a better assessment of hypomanic symptoms, resulting in more correct diagnoses of DMX. PMID- 12551736 TI - Ebselen attenuates reserpine-induced orofacial dyskinesia and oxidative stress in rat striatum. AB - Reserpine-induced orofacial dyskinesia is an alleged animal model of tardive dyskinesia whose pathophysiology has been related to striatal oxidative stress. In the present investigation, the authors examined whether ebselen, an antioxidant organochalcogen with glutathione peroxidase-like activity, changes the behavioral and neurochemical effect of acute reserpine administration. Reserpine injection for 3 days every other day caused a significant increase on the tongue protrusion frequency and ebselen (30 mg/kg ip for 4 days, starting 1 day before reserpine) reversed partially the effect of reserpine (P<.05). Reserpine- and reserpine+ebselen-treated groups displayed an increase in vacuous chewing frequency when compared to control and ebselen-treated groups (P<.05) Reserpine increased the duration of facial twitching and ebselen reversed partially the effect of reserpine (P<.01). Reserpine increased significantly the thiobarbituric acid-reactive species (TBARS) levels, and ebselen reversed the effect of reserpine on TBARS production in rat striatum. The results of the present study clearly indicated that ebselen has a protective role against reserpine-induced orofacial dyskinesia and reversed the increase in TBARS production caused by reserpine administration. Consequently, the use of ebselen as a therapeutic agent for the treatment of tardive dyskinesia should be considered. PMID- 12551737 TI - Alpha2-adrenoceptor antagonist SL 84.0418 reduces the expression of neocortical spike-and-wave spindling episodes in DBA/2J mice. AB - The present work was undertaken to study the effects of alpha(2)-adrenoceptor antagonist 2-(4,5-dihydro-1H-imidazol-2-yl)-1,2,4,5-tetrahydro-2-propyl pyrrolo[3,2,1-hi]-indole hydrochloride (SL 84.0418) on the neocortical spike-and wave spindling episodes (S and W) in the cortical electroencephalogram (ECoG) of DBA/2J mice. Our data indicate that SL 84.0418 (0.1-1.0-10 mg/kg ip) dose dependently reduces the S and W of DBA/2J mice. This effect appears 30 min after drug administration and lasts for the duration of the recording period (240 min). The reducing effect of the SL 84.0418 on the S and W of mice was comparable to that induced by tolazoline (10-20-40 mg/kg ip), a well-known alpha(2) adrenoceptor antagonist, whereas the alpha(2)-adrenoceptor agonist clonidine (0.02-0.1-0.5 mg/kg ip) significantly increased it. The present results indicate that SL 84.0418 administration induces significant reduction on S and W of DBA/2J mice suggesting a possible involvement of noradrenergic system in the development of S and W of DBA/2J mice. PMID- 12551738 TI - Intraventricular 5,7-dihydroxytryptamine lesions disrupt acquisition of working memory task rules but not performance once learned. AB - The serotonergic system is implicated in learning and memory and its disorder, e.g. after 3,4-methylenedioxymethamphetamine (MDMA, "ecstasy") abuse. This study examined the effects of widespread depletion of serotonin (5-HT) using intraventricular injection of 5,7-dihydroxytryptamine (5,7-DHT) on the learning of a working memory task in the dark agouti (DA) rat. The lesion impaired acquisition but not later performance of a nonspatial working memory rule, as measured using nonmatch to sample object recognition in the Y-maze. The lesion had a marginal effect on choice completion times over the course of testing. However, nonspecific effects did not provide a good account of the reduction in choice accuracy as this persisted when completion times were taken into account statistically. Similarly, in a second experiment, the same lesioned rats were slowed in the acquisition of spatial alternation in the T-maze. However, in the open field, there were no comparably long-lasting effects of the serotonergic depletion on line crossings and defecation, only a transient reduction in activity on the first day.Together, these data suggest that the serotonergic system is important in the acquisition of working memory tasks in the rat and that this outcome was unlikely to be the result of nonspecific effects of the lesion. PMID- 12551739 TI - Substance abuse-related P300 differences in response to an implicit memory task. AB - The current study examined electrophysiological measures of cognitive efficiency in alcoholics and controls using a negative priming paradigm derived from DeSchepper and Treisman (J. Exp. Psychol. Learn. Mem. Cog. 22 (1996) 27). In this task, sets of novel shapes were presented: Two overlapping green and red shapes on the left and a single white shape on the right. Participants were instructed to ignore the red shape, but to determine whether the green shape was the same as or different from the white shape. On primed trials, previously red (to be ignored; i.e., irrelevant) shapes became green (relevant) shapes in a second component of the task. Participants who were capable of ignoring irrelevant stimuli were expected to experience more difficulty in the primed condition. Therefore, we predicted that, if alcohol-related impairment in cognitive efficiency is due to inability to respond "appropriately" (i.e., ignore irrelevant stimuli), alcoholics would experience less negative priming than normal controls. Both amplitude and latency of the P300 component of the event related potential were measured in response to each trial condition. Using a 2 x 2 (group x condition) ANOVA with repeated conditions, a significant pattern of group x condition interactions was observed at right frontal, F4 (p=.05) and central parietal, Pz (p=.03) electrode sites on measures of P300 amplitude. A significant group x condition latency interaction was observed at the central parietal electrode, Pz (p=.006). Overall, controls exhibited increased P300 amplitude and latency in response to negatively primed trials. As predicted, alcoholics did not demonstrate this pattern, a finding consistent with alcohol related cognitive inefficiency. PMID- 12551740 TI - A pilot study of plasma caffeine concentrations in a US sample of smoker and nonsmoker volunteers. AB - Even though 85% of adults drink caffeinated beverages daily, very limited studies on plasma caffeine concentration in the US population have been published. Smoking induces cytochrome P450 1A2 (CYP1A2), which is the main enzyme involved in caffeine metabolism. The current naturalistic pilot study explores plasma caffeine concentrations in a US sample, and presents a mathematical model of the relationship between caffeine intake and plasma concentrations for smokers and nonsmokers. Caffeine intake and average plasma caffeine concentrations from morning (7:30-9:30 a.m.) and afternoon (2:00-4:00 p.m.) samples were studied in 69 volunteers (21 smokers and 48 nonsmokers). The mean caffeine intake obtained from caffeinated beverages was 3.02 mg/kg/day, which is similar to the intake in the US population. Almost all subjects in the present sample (99%; 95% confidence interval [CI]: 96-100) had detectable plasma caffeine concentrations. Smokers had significantly higher caffeine intake than nonsmokers. The ratio of concentration/dose of caffeine from caffeinated beverages was approximately four fold higher in nonsmokers (1.33 kgxday/l) than in smokers (0.29 kgxday/l). According to the model, the median plasma caffeine concentration was two- to three-fold higher in nonsmokers for each level of caffeine intake. Our model improves our understanding of the interactions between caffeine and smoking. Additional studies are needed to replicate the model. This model may help epidemiologists to correct for the effects of smoking on caffeine intake and pharmacologists to screen for the activity of CYP1A2. PMID- 12551741 TI - Effects of olanzapine on auditory P300 in schizophrenia. AB - Reduced auditory P300 amplitude generally has been considered to be a trait marker of schizophrenia, independent of antipsychotic treatment and clinical symptoms. However, several seemingly well-conducted studies have found P300 amplitude to be a state marker correlated with clinical symptoms. Recent research on atypical antipsychotics indicate that these medications may alter P300 amplitude as well as having beneficial clinical effects. The objective of the present study was to further elucidate the effects of schizophrenia, symptom severity, and medication status on the P300. The baseline auditory P300 was assessed in unmedicated schizophrenic patients who then were treated with olanzapine for 6 weeks and reassessed. Healthy control subjects were assessed at baseline and again at 6 weeks. Compared to healthy controls, the unmedicated patients' P300s were attenuated and delayed prior to treatment. Subsequent antipsychotic treatment increased the patients' P300 amplitudes without affecting latency. Frontal P300 amplitude was normalized, but parietal P300 amplitude remained below that of healthy controls. Although olanzapine was effective in reducing the patients' symptoms, there were no correlations between symptoms and P300 amplitude or latency either before or after treatment. The results of the present study lend support to the view that P300 amplitude behaves as a trait marker. No evidence is found of a P300 clinical state marker. PMID- 12551742 TI - Effects of acute cholecystokinin infusion on hemispheric EEG asymmetry and coherence in healthy volunteers. AB - This study investigated the effects of continuous slow infusion of cholecystokinin tetrapeptide (CCK-4), a neuropeptide with panicogenic properties, on functional hemispheric differences, as indexed by quantitative electroencephalographic (EEG) asymmetry and coherence measures. Twenty-four adult volunteers (15 females and 9 males) were assigned to infusion with either placebo or CCK-4 in a randomized, double-blind, parallel-group design, with EEG being recorded before and during (10 and 40 min) a 60-min infusion period. No significant treatment differences were observed for absolute EEG power but, compared to placebo, CCK-4 infusion increased asymmetry and reduced coherence of slow-wave activity at midtemporal recording sites. These findings support the contention that functional imbalance of the temporal cortex, perhaps mediated by CCK-4, is involved in panic disorder (PD). PMID- 12551743 TI - Combined treatment with galanthaminium bromide, a new cholinesterase inhibitor, and RS 67333, a partial agonist of 5-HT4 receptors, enhances place and object recognition in young adult and old rats. AB - The present study was designed to investigate whether a combination of a new acetylcholinesterase inhibitor we have synthesized, galanthaminium bromide, and an agonist of 5-hydroxytryptamine(4) receptors, RS 67333, at doses ineffective alone, improves performance in tasks involving place and object recognition memory. Dose responses of each compound were determined in order to select doses without effect alone. Accordingly, young adult rats were injected intraperitoneally with galanthaminium bromide (0.3 mg/kg)+RS 67333 (0.01 mg/kg), and old rats with galanthaminium bromide (0.1 mg/kg for place and 0.3 mg/kg for object recognition)+RS 67333 (1 mg/kg). Drugs were injected before the acquisition phase, immediately after it, or before the retrieval phase to determine the stage of information processing affected by treatments. Doses of galanthaminium bromide and RS 67333, without effect on their own, jointly improved both place and object recognition in young adult rats via an enhancement of acquisition and consolidation information processing. In old rats, the combined treatment enhanced performance by acting on the acquisition processes of place recognition and on the acquisition and consolidation processes of object recognition. These results indicate that combining agents that act on different neuronal targets may be more powerful than either treatment alone, enabling use of lower doses of each compound, thereby attenuating the adverse effects of the individual drugs. A bitherapeutic strategy of this kind might thus be of interest in the treatment of the cognitive deficits related to "normal" or pathological aging. PMID- 12551744 TI - Molecular mechanisms of corticosteroid actions in chronic inflammatory airway diseases. AB - Although corticosteroids have been used for a long time as a very effective therapy of airway inflammatory diseases such as asthma, only recently the molecular basis of their mechanism of action has begun to be elucidated. These hormones exert their biological and pharmacological actions by binding to cytoplasmic receptors that, upon activation, translocate to the nucleus where they interact with specific genomic sequences thus modulating gene expression. However, many glucocorticoid effects responsible for their anti-inflammatory and anti-asthmatic activity take place irrespectively of receptor binding to DNA. In particular, ligand-bound glucocorticoid receptors can repress several different pro-inflammatory genes by physically associating, via protein-protein interactions, with various transcription factors and with the macromolecular complexes implicated in regulation of chromatin structure and function. In this regard, an important role is played by the influences of corticosteroids on the intrinsic histone acetyltransferase and deacetylase functions of coactivators and corepressors, respectively. Furthermore, the signal transduction pathways mediated by mitogen-activated protein kinases are newly recognized, key targets of glucocorticoids. Indeed, these enzymatic cascades are crucially involved in the regulation of gene expression in that they are essential for the activity of a high number of transcription factors. Therefore, the recent advances made in such a rapidly growing research field are providing new insights into the mode of action of corticosteroids, thereby also unveiling novel promising therapeutic strategies directly targeted to the molecular events underlying the inflammatory, immune, and apoptotic processes implicated in the pathogenesis of asthma and other airway diseases. PMID- 12551745 TI - The collagenolytic effects of the traditional Chinese medicine preparation, Han Dan-Gan-Le, contribute to reversal of chemical-induced liver fibrosis in rats. AB - Han-Dan-Gan-Le (HDGL), a Chinese herb preparation composed of Stephaniat tetrandra, Salvia miltorrhiza, Radix paeoniae, Astragalus membranaceus, and Ginkgo biloba, has been used to treat human liver fibrosis. This study was designed to examine the therapeutic effect of HDGL on chemical-induced liver fibrosis in adult Wistar rats. Liver fibrosis was produced in rats by carbon tetrachloride (1.2 ml CCl(4)/kg, 2 times/week, after an initial dose of 5.0 ml CCl(4)/kg, sc), plus a diet of 20% fat, 0.05% cholesterol (continuous) and 30% alcohol in the drinking water ad libitum (every other day) for 8 weeks. HDGL (0.5 and 1.0 g/kg, ig, daily for 6 weeks) was administered to rats 72 hrs after the last dose of CCl(4) to examine its therapeutic effects on chemical-induced liver fibrosis. Upon pathological examination, the HDGL treatment had significantly reversed chemical-induced liver fibrosis and other hepatic lesions. Hepatic collagen accumulation induced by CCl(4) was markedly reduced by HDGL treatment, as evidenced by hepatic collagen content and by immunohistochemical analysis of type-I collagen in liver. HDGL appeared to stimulate the collagenolytic process in the liver, as a 30-50% increase in urinary excretion of hydroxyproline was observed with HDGL treatment as compared to rats only given CCl(4). In conclusion, HDGL can effectively reverse chemically induced liver fibrosis, and this appears to be due, at least in part, to the stimulation of hepatic collagenolysis, resulting in a resolution of hepatic fibrosis. PMID- 12551746 TI - Expression and function of bradykinin receptors in microglia. AB - Expression of bradykinin (BK) receptors and their cellular function were investigated in microglia. Microglial cells were isolated from mixed cultures of cerebrocortical cells from postnatal day 3 Wistar rats. Reverse transcription-PCR (RT-PCR) showed that rat primary microglia express mRNAs for the type 2 bradykinin (B(2)) receptor subtype but not the type 1 (B(1)) receptor subtype under our experimental condition. However, the expression of B(1) receptor was greatly up-regulated after the treatment of microglia with BK for 24 hours. The expression of B(2) receptor in microglia was further confirmed by immunocytochemistry. Membrane currents were measured using whole-cell recording under voltage-clamp conditions. In 14% of patched cells (12/85 cells), BK (100 200 nM) induced an outward current at the holding potential of -20 mV, with oscillations in 2 cases. The BK-induced outward current was transient and desensitized rapidly. TEA inhibited the BK-induced outward current in a dose dependent manner. These results suggest that microglia express B(2) receptors and presumably increase the intracellular Ca(2+) concentration via inositol trisphosphate with the subsequent activation of Ca(2+)-dependent K(+) channels. Our data provide the first evidence that microglia express functional BK receptors and support the idea that microglia play an important role in CNS inflammatory responses. PMID- 12551747 TI - Antioxidant properties of aged garlic extract: an in vitro study incorporating human low density lipoprotein. AB - Oxidation of low-density lipoprotein (LDL) has been recognized as playing an important role in the development and progression of atherosclerotic heart disease. Human LDL was isolated and challenged with a range of oxidants either in the presence or absence of AGE or its diethyl ether extract. Oxidative modification of the LDL fraction using CuSO(4), 5-lipoxygenase and xanthine/xanthine oxidase was monitored by both the appearance of thiobarbituric acid substances (TBA-RS) and an increase in electrophoretic mobility. This study indicates that AGE is an effective antioxidant as it scavenged superoxide ions and reduced lipid peroxide formation in cell free assays. Superoxide production was completely inhibited in the presence of a 10% (v/v) aqueous preparation of AGE and reduced by 34% in the presence of a 10% (v/v) diethyl ether extract of AGE. The presence of 10% (v/v) diethyl ether extract of AGE significantly reduced Cu(2+) and 15-lipoxygenase-mediated lipid peroxidation of isolated LDL by 81% and 37%, respectively. In addition, it was found that AGE also had the capacity to chelate copper ions. In contrast, the diethyl ether extract of AGE displayed no copper binding capacity, but demonstrated distinct antioxidant properties. These results support the view that AGE inhibits the in vitro oxidation of isolated LDL by scavenging superoxide and inhibiting the formation of lipid peroxides. AGE was also shown to reduce LDL oxidation by the chelation of Cu(2+). Thus, AGE may have a role to play in preventing the development and progression of atherosclerotic disease. PMID- 12551748 TI - Protein kinase C-epsilon protects PC12 cells against methamphetamine-induced death: possible involvement of suppression of glutamate receptor. AB - The involvement of PKC isoform in the methamphetamine (MA)-induced death of neuron-like PC12 cell was studied. The death and the enhanced terminal dUTP nick end labeling (TUNEL) staining were inhibited by a caspase inhibitor, z-Val-Ala Asp- (OMe)-CH(2)F (z-VAD-fmk). However, the cell death shows neither morphological nor biochemical features of apoptosis or necrosis. The cell death was suppressed by a protein kinase C (PKC) activator, 12,13-phorbol myristate acetate, but was enhanced by PKC specific inhibitor calphostin C or bisindolylmaleimide, not by PKC inhibitor relatively specific for PKC-alpha (safingol) or PKC-delta (rottlerin). Western blotting demonstrated the expression of PKC-alpha, gamma, delta, epsilon and zeta, of which PKC-epsilon translocated from the soluble to the particulate fraction after MA-treatment. Antisense to PKC epsilon enhanced MA-induced death. A glutamate receptor antagonist MK801 abrogated the cell death, which is reversed by PKC inhibition. These data suggest that PKC-epsilon promotes PC12 cell survival through glutamate receptor suppression. PMID- 12551749 TI - Lemon flavonoid, eriocitrin, suppresses exercise-induced oxidative damage in rat liver. AB - To examine the preventive effect of the lemon flavonoid, eriocitrin (eriodictyol 7-O-rutinoside), on oxidative stress during acute exercise in vivo, levels of N( epsilon )- (hexanoyl)lysine, HEL; o,o-dityrosine, DT; and nitrotyrosine, NT, as oxidative stress markers, were determined by ELISA in livers of trained rats in addition to thiobarbituric acid-reactive substance (TBARS). Eriocitrin administration prior to exercise significantly suppressed the increases in TBARS caused by lipid peroxidation during acute exercise. The contents of HEL, DT, and NT in rat liver increased dramatically by exercise without eriocitrin administration. However, these increases were significantly suppressed by eriocitrin administration before exercise. Moreover, in this study, to clarify whether eriocitrin influences glutathione metabolite system that is considered to be important for a defense against the damage by oxidative stress, the levels of glutathione in rat liver were determined during exercise. The level of reduced glutathione after exercise was maintained by administration of eriocitrin. The increase in the concentration of oxidized glutathione caused by exercise was significantly suppressed by eriocitrin. This result suggested that eriocitrin might play an important role in the control of the change in glutathione redox status in rat liver during exercise. These findings showed that eriocitrin was effective in the prevention of oxidative damages caused by acute exercise-induced oxidative stress. PMID- 12551750 TI - Induction of nitric oxide synthase expression by Withania somnifera in macrophages. AB - Withania somnifera (ashwagandha, Indian ginseng) is an immunostimulant herbal medicine used to improve overall health and prevent diseases, particularly in the elderly. However, the mechanisms underlying its immunostimulant effect is poorly understood. To elucidate the mechanism of Withania somnifera, we investigated the effect of a methanolic extract from the root of Withania somnifera (WS) on nitric oxide (NO) production in J774 macrophages. We found that WS (1-256 microg/ml) produced a significant and concentration-dependent increase in NO production, an effect which was abolished by N(G)nitro-L-arginine methyl ester (L-NAME, 3-300 microM), a non-selective inhibitor of NO synthase (NOS), dexamethasone (10 microM), an inhibitor of protein synthesis and N(alpha-p)-tosyl-L-lysine chloromethyl ketone (TLCK, 0.01-10 microM), an inhibitor of nuclear factor-kappaB (NF-kappaB) activation. Dexamethasone did not have any effect on NO production once NOS had been induced (i.e. 12 h after WS). Moreover, western blot analysis showed that WS increased, in a concentration-dependent fashion, inducible NOS protein expression. These results demonstrate that WS may induce the synthesis of inducible NOS expression likely by acting at transcriptional level. The increased NO production by macrophages could account, at least in part, for the immunostimulant properties of Withania somnifera. PMID- 12551751 TI - Marathon running alters the DNA base excision repair in human skeletal muscle. AB - Reactive oxygen and nitrogen species generated either as products of aerobic metabolism or as a consequence of environmental mutagens, oxidatively modify DNA. Formamidopyrimidine-DNA glycosylase (Fpg) and endonuclease III (endo III) or their functional mammalian homologues repair 7,8-dihydro-8-oxoguanine (8-oxoG) and damaged pyrimidines, respectively, to curb the deleterious effects of oxidative DNA alterations. A single bout of physical exercise can induce oxidative DNA damage. However, its effect on the activity of repair enzymes is not known. Here we report that the activity of a functional homolog of Fpg, human 8-oxoG DNA glycosylase (hOGG1), is increased significantly, as measured by the excision of 32P labeled damaged oligonucleotide, in human skeletal muscle after a marathon race. The AP site repair enzyme did not change significantly. Despite the large individual differences among the six subjects measured, data suggest that a single-bout of aerobic exercise increases the activity of hOGG1 which is responsible for the excision of 8-oxoG. The up-regulation of DNA repair enzymes might be an important part of the regular exercise induced adaptation process. PMID- 12551752 TI - Minocycline blocks 6-hydroxydopamine-induced neurotoxicity and free radical production in rat cerebellar granule neurons. AB - Neurotoxicity induced by 6-hydroxydopamine (6-OHDA) is believed to be due, in part, to the production of reactive oxygen species (ROS). Anti-oxidants by inhibiting free radical generation, protect neurons against 6-OHDA-induced neurotoxicity. In this study, we investigated whether or not minocycline, a neuroprotective compound, could directly protect neurons against 6-OHDA-induced neurotoxicity and inhibit 6-OHDA-induced free radical production in cultured rat cerebellar granule neurons (CGN). We now report that exposure of CGN to 6-OHDA (100 microM) resulted in a significant increase in free radical production with death of 86% of CGN. Pretreatment with minocycline (10 microM) for 2 h prevented 6-OHDA-induced free radical generation and neurotoxicity. Furthermore, minocycline also attenuated H(2)O(2)-induced neurotoxicity. Our results suggest that minocycline blocks 6-OHDA-induced neuronal death possibly by inhibiting 6 OHDA-induced free radical generation in CGN. Both the antioxidative and neuroprotective effects of minocycline may be beneficial in the therapy of Parkinson's disease and other neurodegenerative diseases. PMID- 12551753 TI - Distribution of fatty acids from dietary oils into phospholipid classes of triacylglycerol-rich lipoproteins in healthy subjects. AB - Several studies have suggested that lipoprotein metabolism can be affected by lipoprotein phospholipid composition. We investigated the effect of virgin olive oil (VOO) and high-oleic sunflower oil (HOSO) intake on the distribution of fatty acids in triacylglycerols (TG), cholesteryl esters (CE) and phospholipid (PL) classes of triacylglycerol-rich lipoproteins (TRL) from normolipidemic males throughout a 7 h postprandial metabolism. Particularly, changes in oleic acid (18:1n-9) concentration of PL were used as a marker of in vivo hydrolysis of TRL external monolayer. Both oils equally promoted the incorporation of oleic acid into the TG and CE of postprandial TRL. However, PL was enriched in oleic acid (18:1n-9) and n-3 polyunsaturated fatty acids (PUFA) after VOO meal, whereas in stearic (18:0) and linoleic (18:2n-6) acids after HOSO meal. We also found that VOO produced TRL which PL 18:1n-9 content was dramatically reduced along the postprandial period. We conclude that the fatty acid composition of PL can be a crucial determinant for the clearance of TRL during the postprandial metabolism of fats. PMID- 12551754 TI - PPAR-gamma, TNF-alpha messenger RNA levels and lipase activity in the pregnant and lactating rat. AB - Dramatic alternations in maternal metabolism occur during gestation and lactation, especially glucose and fat metabolism. For example, in rats, the amount of body fat mass increases during gestation, then decreases just prior to delivery, and remains low after parturition. To investigate the factors involved in such changes in maternal fat mass, messenger ribonucleic acid (mRNA) levels of adipocytokines, peroxisome proliferator-activated receptor-gamma (PPAR-gamma) and tumor necrosis factor-alpha (TNF-alpha), were examined in the intraabdominal adipose tissue of non-pregnant rats, pregnant rats and postpartum rats. We also examined the issue of whether apoptosis, which could be promoted by PPAR-gamma and TNF-alpha, is involved in any of the changes in maternal fat mass The activity of lipoprotein lipase (LPL) and hormone sensitive lipase (HSL) in adipose tissue was also measured. PPAR-gamma and TNF-alpha mRNA levels remained constant during the gestational and postpartum periods. Apoptosis was not detected at any time as evidenced by DNA laddering and in situ staining. LPL activity was significantly increased at day 5 and remained elevated until day 14 of gestation. HSL activity was significantly increased at day 10 of gestation and then decreased after delivery, at day 10 of lactation. In conclusion, during the gestational and postpartum period of rats, changes in maternal fat mass did not directly correlate with the levels of expression of PPAR-gamma and TNF-alpha mRNA. Apoptosis also does not appear to influence on fat mass change. The changes in LPL and HSL activities during gestation suggest that these enzymes might be regulators of maternal adipose tissue level. PMID- 12551755 TI - Alcohol, injury, and cellular immunity. AB - It is widely accepted that alcohol exposure is a causative factor in the occurrence of burn or other traumatic injury. It is less well known that individuals who have consumed alcohol before sustaining an injury suffer from increased morbidity and mortality compared with the morbidity and mortality of non-alcohol-consuming subjects with similar injuries. Complications due to bacterial infection are the most common burn sequelae in injured patients and are frequently associated with depressed immunity. Independently, alcohol exposure and injury have been shown to influence cellular immunity negatively. These changes in immunity are closely linked to injury- or alcohol-induced alterations in the cytokine milieu in both clinical studies and animal models. Not surprisingly, the combination of insult of alcohol exposure and burn injury results in immune suppression that is greater in magnitude and duration compared with either insult alone. The combined effects of alcohol and injury on immunity have been examined in a limited number of studies. However, results of these studies support the suggestion that altered cytokine production is an integral part of the immune dysregulation and increased mortality that is observed. In particular, the increased presence of macrophage-derived mediators observed after burn or alcohol exposure alone seems to be synergistically increased in a combined injury model. Although more research is needed, it is likely that therapeutic modalities that include manipulation of cytokine networks to boost cellular immunity may improve outcome for patients who sustain injuries subsequent to consuming alcohol. PMID- 12551756 TI - Effects of maternal ethanol consumption on hematopoietic cells in the rat fetal liver. AB - During development, there are many factors to be considered in studying the efficacy of the hematopoietic system to provide the immune system with adequate numbers of functional cells within the immune repertoire. Hematopoietic cells must be able to develop, proliferate, and emigrate from the hematopoietic fetal liver to other tissues of the body, such as the thymus and bone marrow. There is evidence that ethanol consumption causes immune deficiencies in adults and that maternal ethanol consumption causes immune deficiencies in children, both with correlative effects on cells and cytokinetic regulators. Therefore, the ability of the hematopoietic system to seed the immune system may be jeopardized by maternal ethanol consumption. In this study, test groups included female rats (1). fed a Lieber-DeCarli ethanol liquid diet, (2). pair-fed a control liquid diet, or (3). fed standard laboratory chow. Livers were removed from fetuses 18 and 21 days postconception (dpc) and analyzed by immunophenotyping and flow cytometry. There was a 22% and 38% decrease in fetal body weight and a 25% and 30% decrease in fetal liver weights between ethanol-fed and pair-fed groups at 18 and 21 dpc, respectively. At 18 dpc, fetal body and liver weights of the pair-fed animals were also significantly reduced to those of the chow-fed group. However, by 21 dpc, both body and liver weights of the two control groups were not statistically different. The effects of maternal ethanol consumption on the distribution of hematopoietic cells were characterized by using monoclonal antibodies (mAbs) anti-CD43 (a progenitor hematopoietic cell marker) versus labeling with anti-Vbeta8.2 (pre-T cells), anti-B220 (pre-B cells), and anti-NKR P1A (pre-natural killer cells). With the use of flow cytometric analysis, at 18 dpc ethanol-exposed fetuses showed 40% and 62% decreases in B220(+) and Vbeta8.2(+) cells, respectively, versus findings for pair-fed controls, with no significant change in NKR-P1A(+) cells. At 21 dpc, ethanol-exposed fetuses showed a 46% decrease in B220(+) cells among the CD43(+) cells, with a 50% increase in Vbeta8.2(+) cells. Observed alterations in gross fetal body and liver weights, together with modifications in the percentage of hematopoietic cells in the fetal liver after maternal ethanol consumption, strongly support the suggestion that the ability of the hematopoietic system to impart a repertoire of cells that are functional in the regulatory and effector mechanisms of the immune process may be compromised. PMID- 12551757 TI - Role of different nicotinic acetylcholine receptors in mediating behavioral and neurochemical effects of ethanol in mice. AB - Ethanol and nicotine are the most abused drugs, and it is well known that co abuse of ethanol and nicotine is frequent in human beings. We have previously obtained results indicating that the ethanol-induced stimulation of both the mesolimbic dopamine system and locomotor activity may involve activation of central nicotinic acetylcholine receptors (nAChRs), especially those located in the ventral tegmental area. Different subpopulations of nAChRs have been identified, and, in the present series of experiments, we have studied the effects of various nAChR antagonists on the stimulation of dopamine overflow in the nucleus accumbens and on locomotor activity induced by ethanol in male mice. Ethanol (2.0 g/kg, i.p.) enhanced dopamine overflow in the nucleus accumbens by approximately 40%, measured by means of in vivo microdialysis in awake, freely moving mice. Mecamylamine (negative allosteric modulator of nAChR; 2.0 mg/kg, i.p.) blocked the ethanol-induced stimulation of both locomotor activity and accumbal dopamine overflow. Methyllycaconitine citrate (alpha(7) antagonist; 2.0 mg/kg, i.p.) and dihydro-beta-erythroidine (competitive and selective alpha(4)beta(2) antagonist; 0.5 mg/kg, s.c.), in doses that had no marked effects per se, did not significantly reduce the behavioral and neurochemical stimulation caused by ethanol. The present results support the suggestion that the stimulatory effects of ethanol on locomotor activity and dopamine release do not involve the alpha(4)beta(2) or alpha(7) subunit compositions of the nAChR and that the effects of mecamylamine are mediated through a site not directly associated with the alpha(4)beta(2) or alpha(7) nAChR subunits. PMID- 12551758 TI - Effect of naltrexone on oral consumption of concurrently available ethanol and cocaine in the rat. AB - Comorbid abuse of and dependency on multiple drugs is a common occurrence clinically. We have developed an animal model that provides rats with the opportunity to choose, through oral consumption, between concurrently available ethanol and cocaine with water also available. This provides the ability to screen for the effectiveness of potential pharmacotherapeutic agents on the baseline consumption of both drugs. We used this animal model to evaluate the effects of naltrexone, at doses of 0, 1.0, 3.0, and 10.0 mg/kg, on concurrent oral consumption of ethanol and cocaine solutions. Naltrexone at all doses significantly reduced both consumption of and preference for ethanol. Consumption of both cocaine and water was unaffected by naltrexone, supporting the suggestion that the effects of naltrexone were selective for ethanol. These findings support the suggestion that ethanol and cocaine act on different central reward pathways. The implications of these findings for the clinical use of naltrexone in populations with comorbid ethanol and cocaine abuse are discussed. PMID- 12551759 TI - Extremely long recovery time for the sedative effect of clonidine in male type 1 alcohol-dependent subjects in full sustained remission. AB - The possible relation between alpha-2-adrenoceptor function-as assessed by changes in systolic and diastolic blood pressure and heart rate, as well as level of sedation, after administration of clonidine (2.0 microg/kg, i.v.)-and length of time of alcohol dependence or duration of remission was investigated in 17 male subjects with alcohol dependence in full sustained remission. Six healthy males were used as control subjects. The clonidine-induced scores for level of sedation were found to correlate with duration of time in remission (r = 0.60; P <.02). Median split of duration of remission revealed that subjects with short term (2 +/- 1 years) duration of remission had significantly lower scores for clonidine-induced level of sedation than the scores for both subjects with long term (12 +/- 5 years) duration of remission (P <.004) and control subjects (P <.02). There was also a significant correlation between duration of remission and values for clonidine-induced reduction of systolic blood pressure (r = 0.51; P <.05). Results indicate an extremely long recovery period in some aspects of alpha-2-adrenoceptor function, especially for clonidine-induced increase in level of sedation, with a normalization time of 4 to 5 years. PMID- 12551760 TI - Maintaining steady state arterial alcohol levels in rats by using a physiologically based pharmacokinetic model. AB - An intravenous method of alcohol administration that maintains arterial alcohol concentrations (AACs) in rats at a prescribed level for a prolonged period was previously described. This method produced steady state AACs between 30 and 180 min after the start of the infusion, but it resulted in substantial overshoots in AAC initially. The present study was designed to achieve target AACs close to steady state more quickly while minimizing any overshoot. A physiologically based pharmacokinetic (PBPK) model of alcohol distribution and elimination was developed for male Wistar rats. Body weight was used to compute individualized infusion rate profiles that would achieve steady state AACs of 75, 150, and 250 mg%. Rats were chronically implanted with cannulae in the jugular vein (for alcohol infusion) and carotid artery (for blood sampling). Alcohol was administered according to the individualized infusion rate profiles. Blood was collected at intervals for AAC determination. The PBPK model-based infusion profiles achieved target AACs 5 min after the start of the infusion and maintained the AACs for 2 h. The AACs deviated an average of 4.9%, 5.1%, and 5.9% from target at the 75, 150, and 250 mg% levels, respectively. Through the application of a PBPK model, it is possible to achieve target AACs close to steady state more quickly in male Wistar rats and to minimize any overshoot in AAC. The PBPK model-based method seems to be improved over the earlier method. Maintaining steady state AACs in rats is useful for studies in which fluctuating alcohol levels may confound experimental results. PMID- 12551762 TI - Industrial applications of sonochemistry in the syntheses of vitamin-building blocks. AB - In this article, the application of ultrasound in syntheses of vitamins and fine chemicals is presented. Based on selected examples, the use of ultrasound in dehydration, isomerisation, and decarboxylation reactions is described. PMID- 12551761 TI - Morphologic and neurotoxic effects of ethanol vary with timing of exposure in vitro. AB - Results of investigations with animal models of fetal alcohol syndrome (FAS) seem to indicate that neuronal vulnerability to ethanol-induced cell death may be correlated with specific developmental events. In the present study, we sought to test this observation in a cell culture model of neuronal development in which morphogenesis as well as survival could be assessed. Using embryonic rat hippocampal pyramidal neurons in primary cultures, we compared the sensitivity of neurons to ethanol added, at 400 mg/dl, to the medium at different times relative to the development of axons and dendrites. Quantitative morphometric analysis was performed by using phase contrast at 12 h (0.5 day) and 24 h (1 day), or fluorescence microscopy after microtubule-associated protein-2 (MAP2) immunostaining at 6 and 14 days. Survival was assessed by counting the number of neurons per unit area of the substrate at 14 days. Addition of ethanol 1 day after plating, when most neurons had developed an axon, had no effect on survival up to 14 days in vitro, but resulted in significantly shorter, less branched dendrites than observed when ethanol was added 2 h after plating. Despite the shorter duration of ethanol exposure, the addition of ethanol on day 6, after rapid growth of dendrites and synapses had begun, resulted in loss of all but about one third of the neurons by 14 days. This supports the suggestion that increased neuronal vulnerability to the morphoregulatory effects of ethanol is correlated with the establishment of polarity, but that the sensitivity of neurons to the cytotoxic effects of ethanol occurs later, when dendrites and synapses are rapidly forming. PMID- 12551763 TI - Preparation of highly dispersed CuO catalysts on oxide supports for de-NO(x) reactions. AB - CuO based catalysts dispersed on silica-alumina supports at low (0.56 wt.%) and high (13 wt.%) Al(2)O(3) content were prepared by adsorption method with or without ultrasound treatment. The catalysts obtained were studied in their bulk (atomic absorption, X-ray diffraction, temperature programmed reduction) and surface (N(2) adsorption and X-ray photoelectron spectroscopy) properties. Significant differences between the series of catalysts prepared over the two supports in terms of size of the CuO aggregates and of their redox properties were evidenced. All the catalysts were tested in the selective catalytic reduction of NO(x) using C(2)H(4) as reducing species (HC-SCR process) in highly oxidant atmosphere. The CuO-catalysts prepared using ultrasounds were the most active. Moreover, they displayed a peculiar activity being able to activate NO both by reducing it to N(2), in larger extent, and by oxidizing it to NO(2). PMID- 12551764 TI - Electrochemical detection of bubble oscillation. AB - The interaction of a tethered bubble with sound is demonstrated using novel electrochemical characterisation technology. A 25 microm diameter microelectrode, positioned close to the gas/liquid interface is used to monitor the motion of the bubble wall as a function of time in the presence and absence of sonic irradiation. Evidence for 'breathing' mode oscillation of the bubble and its effect on mass transfer to the microelectrode is presented. PMID- 12551765 TI - Ultrasonic-induced nucleation of ice in water containing air bubbles. AB - Cavitation induced by ultrasonic vibrations can cause nucleation of ice in supercooled water. In this study, the time required for ultrasonic-induced nucleation of ice was measured for water containing two different size distributions of air bubbles. When the water was supersaturated with air bubbles, there was a time lag of about 0.5 s between the onset of ultrasonic irradiation and the onset of ice nucleation, and the probability of ice nucleation was unusually high within 0.5-1.1 s after the onset of ultrasonic irradiation. These results cannot be explained by conventional models alone, in which the collapse of a cavitation bubble triggers the nucleation of ice. Secondary effects appear to also influence ice nucleation. PMID- 12551766 TI - Preparation of conducting polyaniline colloids under ultrasonication. AB - The effects of ultrasonication on the chemical polymerization of aniline leading to the formation of conducting polyaniline colloids were examined. The formation rate of the colloids was significantly increased under ultrasonication. Furthermore, it was also observed that the morphological structure of the colloids thus prepared was greatly affected by the sonication. The polyaniline colloids were further characterized by a range of techniques including electric resistance meter, gel permeation chromatography, FT-IR and cyclic voltammetry. It is noteworthy that the application of ultrasound to the polymerization resulted in a marked increase in the doping level, which reflected to the high electroconductivity of polyaniline colloids. PMID- 12551767 TI - Dose-dependent inhibition of ultrasound-induced cell killing and free radical production by carbon dioxide. AB - Previous studies have shown that if a solution for cell suspension is saturated with CO(2), ultrasound-induced in vitro cell killing and free-radical production are inhibited. However, the dose dependency of this observation has not been explored. Here, we used NaHCO(3) and HCl to produce a predictable concentration of CO(2) within the culture medium. Using 1 MHz continuous wave 4 W/cm(2) ultrasound, we sonicated U937 cells suspension for 1 min at 37 degrees C with CO(2) at different concentrations. At 2 mM, reduced cell killing was observed that further decreased with increasing CO(2) concentration until 100% protection was attained at 20 mM. Ultrasound-induced free-radical production was significantly decreased at 1 mM and became undetectable at 2 mM CO(2). This finding shows that CO(2)-mediated inhibition is concentration dependent and that the threshold for free-radical production is one order of magnitude higher than the threshold for cell killing induced by ultrasound. In addition, it also cautions researchers when adding acids and acid-based agents to a culture medium, which almost always contains NaHCO(3), in experiments related to the bioeffects of ultrasound. PMID- 12551768 TI - Studies on the use of power ultrasound in leather dyeing. AB - Uses of power ultrasound for acceleration/performing the chemical as well as physical processes are gaining importance. In conventional leather processing, the diffusion of chemicals through the pores of the skin/hide is achieved by the mechanical agitation caused by the paddle or drumming action. In this work, the use of power ultrasound in the dyeing of leather has been studied with the aim to improve the exhaustion of dye for a given processing time, to reduce the dyeing time and to improve the quality of dyed leather. The effect of power ultrasound in the dyeing of full chrome cow crust leather in a stationary condition is compared with dyeing in the absence of ultrasound as a control experiment both in a stationary as well as conventional drumming condition. An ultrasonic cleaner (150 W and 33 kHz) was used for the experiments. Actual power dissipated into the system was calculated from the calorimetric measurement. Experiments were carried out with variation in type of dye, amount of dye offer, temperature and time. The results show that there is a significant improvement in the percentage exhaustion of dye due to the presence of ultrasound, when compared to dyeing in absence of ultrasound. Experiments on equilibrium dye uptake carried out with or without ultrasound suggest that ultrasound help to improve the kinetics of leather dyeing. The results indicate that leathers dyed in presence of ultrasound have higher colour values, better dye penetration and fastness properties compared to control leathers. The physical testing results show that strength properties of the dyed leathers are not affected due to the application of ultrasound under the given process conditions. Apparent diffusion coefficient during the initial stage of dyeing process, both in presence and in absence of ultrasound was calculated. The values show that ultrasound helps in improving the apparent diffusion coefficient more for the difficult dyeing conditions such as in the case of metal complex dyes having bigger aggregate size compared to less difficult dyeing conditions. PMID- 12551769 TI - Sonolysis of metal beta-diketonates in alkanes. AB - The kinetics of metal beta-diketonates sonolysis was studied in hexadecane solutions using a UV/VIS spectrophotometric technique. The following complexes were prepared and studied: Cu(HFAA)(2), Cu(DPM)(2), Fe(ACAC)(3), Ni(DPM)(2), Er(DPM)(3), Nd(DPM)(3), Th(DPM)(4), UO(2)(BTFA)(2).TOPO, and Np(HFAA)(4), where HHFAA is hexafluoroacetylacetone, HDPM is dipivaloylmethane, HACAC is acetylacetone, HBTFA is benzoyltrifluoroacetone, and TOPO is trioctylphosphine oxide. Sonolysis was performed under the following conditions: ultrasonic frequency 22 kHz, intensity of ultrasound 3-5 Wcm(-2), temperature 70-92 degrees C, Ar atmosphere. The kinetic behavior of the studied complexes are interpreted using a two-site model of the sonochemical processes. In the case of metal beta diketonates with high vapor pressure the sonochemical reactions tend to occur in the gaseous phase of the cavitating bubbles. The sonolysis of less volatile complexes first occur in the liquid reaction zone surrounding the bubbles. Sonication of the studied complexes results in the formation of X-ray amorphous products consisted of a mixture of metal beta-diketonates partial degradation products. Heating of as-prepared sonication products in air yields nanocrystalline oxides of corresponding metals. PMID- 12551770 TI - Sonochemical degradation of phenol in water: a comparison of classical equipment with a new cylindrical reactor. AB - Cavitation due to ultrasonic waves produces highly reactive oxidising species in water. As a result, it can be used to oxidise organic pollutants such as aromatic compounds in dilute aqueous solutions. Recent studies have demonstrated that reactors operating in the high frequency range (e.g. 500 kHz) are more efficient than reactors working at lower frequency (20 kHz) for the destruction of these kinds of contaminants. Our study describes the degradation of phenol with the help of a cylindrical ultrasonic apparatus that operates at 35 kHz (Sonitube SODEVA). To date, the use of this type of reactor has not been reported. The reaction rates thus obtained were compared to those obtained at the same ultrasonic power (50 W) with more classical devices operating at 20 and 500 kHz. The general result is that in aqueous solution, the rate of phenol destruction is higher at 500 kHz than at 35 or 20 kHz. Addition of hydrogen peroxide and copper sulphate to the medium provides a different oxidative system that proceeds more efficiently at 35 kHz; the time of destruction was about one-third of the time needed at 500 kHz. It was also observed that the intermediate organic compounds are eliminated much faster at 35 kHz in comparison with the two frequencies. The observation of such different behaviour is not necessarily a pure frequency effect, but can be due to a response to other parameters such as the acoustic field and intensity. PMID- 12551771 TI - Effects of ultrasound on adsorption-desorption of p-chlorophenol on granular activated carbon. AB - The aim of this work is the evaluation of the effects of ultrasound on p chlorophenol adsorption-desorption on granular activated carbon. Adsorption equilibrium experiments and batch kinetics studies were carried out in the presence and the absence of ultrasound at 21 kHz. Results indicate that the adsorption of p-chlorophenol determined in the presence of ultrasound is lower than the adsorption observed in the absence of ultrasound. Desorption of p chlorophenol from activated carbon with and without the application of ultrasound was studied. The desorption rates were favoured by increased ultrasound intensity. This rise is more noticeable as temperature increases. The addition of ethanol or NaOH to the system causes an enhancement of the amount of p chlorophenol desorbed, especially in the presence of ultrasound. A synergetic enhancement of the desorption rate was observed when ultrasonic irradiation was coupled with ethanol chemical regeneration. PMID- 12551772 TI - The Michael reaction catalyzed by KF/basic alumina under ultrasound irradiation. AB - The Michael reaction of chalcones as acceptors with various active methylene compounds such as diethyl malonate, nitromethane, cyclohexanone, ethyl acetoacetate and acetylacetone as donors catalyzed by KF/basic alumina results in adducts in high yield within a shorter time under ultrasound irradiation. PMID- 12551774 TI - Nutrient-gene interactions in early pregnancy: a vascular hypothesis. AB - It is hypothesized that the following periconceptional and early pregnancy nutrient-gene interactions link vascular-related reproductive complications and cardiovascular diseases in adulthood: (1) Maternal and paternal genetically controlled nutrient status affects the quality of gametes and fertilization capacity; (2) The embryonic genetic constitution, derived from both parents, and the maternal genetically controlled nutrient environment determine embryogenesis and fetal growth; (3) Trophoblast invasion of decidua and spiral arteries is driven by genes derived from both parents as well as by maternal nutritional factors; (4) Angiogenesis, vasculogenesis and vascular function are dependent on the genetic constitution of the embryo, derived from both parents, and the maternal genetically controlled nutritional environment.Early intra-uterine programming of vessels may concern the same (in)dependent determinants of vascular-related complications during pregnancy and cardiovascular diseases in later life. PMID- 12551775 TI - The role of leptin in fertility. AB - The relationship between metabolism and reproduction remains a mystery in female endocrinology. Such substances as insulin, amino acids and IGFBP-I have been proposed as signals of body mass fat on the genital axis. Today this role is claimed by leptin, a protein hormone decoded from the obesity gene and is secreted exclusively from adipose tissue. This hormone acts on the central nervous system (CNS) to result in the suppression of food intake and increase in energy consumption. What is more, it also influences the capacity for reproduction. This paper reports findings with regard to the factors influencing the secretion of leptin and identification of the leptin's hormonal receptors. Particular emphasis was placed on the relationship between secretion of leptin and disturbances in menstruation, the anticipated role of this hormone in the pathogenesis of the polycystic ovarian syndrome (PCOS) and its effects on the reproductive capacity. PMID- 12551776 TI - Comparison of neonatal outcome including cerebral palsy between abruptio placentae and placenta previa. AB - OBJECTIVE: Our purpose was to evaluate the neonatal prognosis after abruptio placentae and placenta previa during pre-term gestation. STUDY DESIGN: A case control study was performed using a logistic regression model. A poor outcome was defined as neonatal death occurring before hospital discharge or a diagnosis of cerebral palsy. RESULTS: A poor outcome was more frequent in cases of abruptio placentae (11/42, 26.2%) than in placenta previa (2/72, 2.8%) and pre-term labor (1/120, 0.8%). The difference was mainly due to the incidence of cerebral palsy. A significant association of abruptio placentae (odds ratio (OR) 61.0, 95% confidence interval (CI 3.4-1084), delivery at <31 weeks of gestation (OR 19.0, CI 2.8-128.8), and low Apgar score (<7) at 5min (OR 70.8, CI 16.5-304.9) with increased risk of poor outcome was found in the logistic regression model that controlled for confounding effects. In abruptio placentae, a low Apgar score (<7) at 5min (OR 19.8, CI 2.0-197.8) was associated with increased risk of poor outcome in the logistic regression model. CONCLUSION: From the standpoint of poor perinatal outcome including cerebral palsy, abruptio placentae was the most significant clinical entity in pre-term gestation. PMID- 12551777 TI - Forceps delivery and the use of synthetic opioid analgesia during epidural anaesthesia. AB - OBJECTIVE: To verify whether the use of sufentanil during obstetric epidural anaesthesia (EA) was accompanied by an increase in the rate of instrumental extraction. STUDY DESIGN: We made a retrospective comparison of deliveries that occurred during two 3-year periods (1993-1995, and 1997-1999), before and after the introduction of sufentanil into our protocol for epidural anaesthesia. RESULTS: The first period covered 4694 deliveries compared with 5310 in the second; for these periods, the EA rate rose from 48.55 to 63.36% (P=0.0001). The rates of uncomplicated vaginal delivery, forceps delivery and caesarian section over the two periods were 70.24, 12.76 and 17% versus 67.08, 14.41 and 18.51%, respectively: the differences were statistically significant. The indications for the use of forceps did not differ during the two periods. During vaginal delivery, the rate of forceps use as a function of EA administration/non administration was remarkably constant over the two periods: 25.8% with EA versus 6.2% without EA in the first period compared with 24.9 and 6.2% in the second period. The forceps delivery rate was unaffected by the fact that EA was carried out by an anaesthetist specialised in obstetrics. CONCLUSION: The addition of sufentanil to our protocol for EA has not altered the practice of instrumental extraction in our department. With EA, the rate of forceps delivery is multiplied by 4, irrespective of the protocol used. PMID- 12551778 TI - Evaluation of a training program for healthcare professionals about breast feeding. AB - OBJECTIVE: To assess the impact of a continuing medical education program based on the WHO's 10 steps to successful breast-feeding. STUDY DESIGN: An observational before-and-after study at a teaching hospital. Data for two random samples of 50 women before and 50 after the intervention were collected from medical records and completed by a mail questionnaire. RESULTS: Seventy-six percent of mothers initiated breast-feeding in the two samples. The median duration of breast-feeding was 12 weeks and did not differ between the before and after groups. The percentage of newborns separated from their mother more than 4h a night decreased substantially after the intervention (13% versus 52% before the intervention, P<0.01). In-hospital formula provision also decreased after the intervention (63% of newborns versus 82% before intervention, P=0.07). CONCLUSION: This pilot study has enabled authors to document the feasibility of evaluating the impact of continuing education of maternity ward staff. PMID- 12551779 TI - Nifedipine-induced changes in body composition in hypertensive patients at term. AB - BACKGROUND: There are few studies on the effect of nifedipine on fluid homeostasis during pregnancy. OBJECTIVE: To assess the variation of body composition at term in patients with normal pregnancy and patients with gestational hypertension (GH) treated with nifedipine. METHODS: Seventeen healthy patients with singleton pregnancies and 12 non-proteinuric patients with gestational hypertension treated with nifedipine were enrolled consecutively from our University Hospital from January 2000 to January 2001. Two evaluations were performed (anthropometric measurements, bioelectric impedance analysis (BIA) at 5, 50 and 100kHz and hematocrit), the first at 36.4+/-3.2 weeks' gestation, the second at 2.1+/-1.6 days of puerperium. The BIA measurements included: fat mass (FM, kg), fat-free mass (FFM, kg), extra-cellular water (ECW, Lt), intra-cellular water (ICW, Lt), total body water (TBW, Lt). Values were analyzed as mean+/-S.D. and statistical analysis was performed by using the Student's t-test for unpaired data. RESULTS: Birthweights were significantly lower in the gestational hypertensive patients (3472.7+/-266.4g versus 2735.8+/-575.9g, P<0.001). At the first study time, systolic and diastolic blood pressure were significantly higher in the hypertensive patients than in control group, notwithstanding the therapy. Interestingly we found pre-delivery and post-delivery TBW higher in gestational hypertensive patients versus the control patients (44.48+/-9.12Lt versus 36.60+/ 5.07Lt, P<0.001 and 40.69+/-4.31Lt versus 37.30+/-4.35Lt, P=0.016, respectively). TBW loss after delivery was greater among hypertensive patients (3.79+/-4.81Lt versus -0.70+/-0.72Lt, P<0.001). CONCLUSIONS: Bioelectrical impedance analysis is a practical tool in the follow-up of anti-hypertensive therapy at term gestation. Moreover, nifedipine increases TBW in gestational hypertensive patients, reverting the pathophisiological pattern, yet described elsewhere. PMID- 12551780 TI - Amniotic fluid glucose at the time of genetic amniocentesis: correlation with duration of pregnancy and birthweight. AB - OBJECTIVE: To test whether amniotic fluid (AF) glucose levels in 15-18 weeks of pregnancy can predict birthweight and duration of pregnancy. STUDY DESIGN: In this prospective study, 66 non-diabetic pregnant women underwent amniocentesis between 15 and 18 weeks of pregnancy and glucose levels were measured immediately by using the enzymatic method of hexocinase. They were all followed up and delivered in our department and neognological parameters were recorded. RESULTS: AF glucose values ranged from 12 to 69mg/dl (mean 44.2mg/dl, S.D. 11.6mg/dl). No linear relationship was found between AF glucose and either birthweight or duration of pregnancy. CONCLUSION: AF glucose values between 15 and 18 weeks of pregnancy did not correlate with either birthweight or duration of pregnancy. A single measurement of this value cannot be used for the prediction of the factors studied. PMID- 12551781 TI - Decreased levels of fibrinogen in very high parity women. AB - OBJECTIVES: To evaluate coagulation parameters in healthy pregnant grand grand multiparous women. STUDY DESIGN: Prothrombin time (PT), activated partial thromboplastin time (PTT), fibrinogen, D-dimer, albumin and blood lipids were prospectively determined in 50 pregnant grand grand multiparas (> or =9 deliveries) and 50 age-matched pregnant women with low parity (< or =3 deliveries). RESULTS: The average concentrations of fibrinogen in the grand grand multiparas were 4.0+/-0.6 as compared to 4.7+/-0.7 g/l in the control group (P<0.001). In both groups D-dimer levels were >1000 ng/ml and the concentrations of albumin, triglycerides, cholesterol, high-density lipoprotein (HDL) were not significantly different. CONCLUSIONS: In healthy pregnant grand grand multiparous women fibrinogen levels are lower than in age-matched women with low parity. These changes are not directly associated with lipids metabolism during pregnancy. PMID- 12551782 TI - Imidazoline receptors in the human umbilical cord. AB - Imidazoline binding sites (IBS) are now accepted as being receptors, however, their physiological functions are not yet clearly understood. Previously, the authors demonstrated that the density of IBS in the human placenta significantly increased throughout gestation. The present study was performed for the identification of imidazoline receptors in the human umbilical cord. PMID- 12551783 TI - Postterm with favorable cervix: is induction necessary? AB - OBJECTIVE: To study the cesarean rate between expectant management and immediate induction in the otherwise uncomplicated postterm pregnancy with favorable cervix. STUDY DESIGN: A total of 249 women with uncomplicated pregnancies at 41 weeks plus 3 days (290 days) with favorable cervix (Bishop score > or =6) were randomized to either expectant management (n=125) or immediate induction of labor (n=124). The women in the induction group were sent to labor ward for induction by artificial rupture of membranes (ARM) and/or oxytocin infusion. The women with expectant management had nonstress test (NST) and amniotic fluid index (AFI) performed once a week and twice a week after 43 weeks of gestation until spontaneous labor. RESULTS: The cesarean rate was not different between expectant management and immediate induction (21.6% versus 26.6%; P=0.36). Ninety-five percent of the expectant group delivered within 1 week after enrollment, and all of them delivered within 9 days after randomization. Maternal and fetal complications in both groups were not different. There was also no difference in the mean birth weight (P=0.24) and the frequency of macrosomia (birth weight > or = 4000 g) between the two groups (P=0.23). CONCLUSION: Cesarean section rate between expectant management and immediate induction in the otherwise uncomplicated postterm pregnancy with favorable cervix was not different. Due to the very low adverse perinatal outcome, both expectant management and immediate induction are acceptable. PMID- 12551784 TI - Placental influences on the rate of labour progression: a pilot study. AB - Regional variations in intrapartum myometrial contractility were assessed using ultrasound. In 10 women with normal labours, the myometrial thickness increased in all areas during contractions, whilst in 10 women with slowly progressing labour, the retro-placental myometrium thinned during contractions. Locally acting placental tocolytics may be a cause of dysfunctional labour. PMID- 12551785 TI - Mid-trimester amniotic fluid angiogenin, lactate dehydrogenase and fibronectin in the prediction of preterm delivery. AB - OBJECTIVE: Our purpose was to compare angiogenin, lactate dehydrogenase (LDH) and fibronectin levels in mid-trimester amniotic fluid of patients with preterm and term deliveries and to find out their predictive values for preterm birth. STUDY DESIGN: A prospective cohort study was conducted in 55 pregnancies with singleton gestations that underwent amniocentesis at 15-20 weeks for standard genetic indications. Amniotic fluid angiogenin, lactate dehydrogenase and fibronectin levels were measured by enzyme-linked immunosorbent assay (ELISA), radial immundiffusion technic and automated analyzer, respectively. RESULTS: Five patients delivered preterm, five developed signs or symptoms of threatened preterm labor and 45 had term delivery after an uneventful pregnancy. Demographic data were not significantly different. Amniotic fluid angiogenin and lactate dehydrogenase levels were significantly higher in patients with preterm than term deliveries (P<0.001 and 0.02, respectively). Receiver-operator characteristic curve analysis showed that the amniotic fluid angiogenin had the best screening efficiency in predicting preterm delivery. An angiogenin level of 35ng/ml was the optimal cut-off value for the prediction of preterm delivery, with a sensitivity of 100% and specificity of 91%. CONCLUSION: Second-trimester angiogenin is found to be quite effective in the prediction of preterm delivery. Preexisting intrauterine ischemia may be an important risk factor for preterm delivery and already be present in the early mid-trimester. PMID- 12551787 TI - Soluble intercellular adhesion molecule 1 in the peritoneal fluid of patients with endometriosis correlates with the extension of peritoneal implants. AB - OBJECTIVE: To investigate the correlation of the concentrations of soluble intercellular adhesion molecule 1 (sICAM-1) in the peritoneal fluid (PF) with the extent of peritoneal endometriotic lesions and with the presence of red lesions (RLs). STUDY DESIGN: Sixty-seven patients with endometriosis and 19 controls with normal pelvis had PF samples collected during laparoscopy. RESULTS: The concentrations of sICAM-1 in the PF of patients and controls were not different. Patients with peritoneal implant scores 4 or more had higher concentrations of sICAM-1 in the PF than those with implant scores less than 4 (P=0.018) and controls (P=0.031). No significant difference was found in sICAM-1 levels in patients with and without RLs. No correlation was detected between sICAM-1 and interleukin 1beta (IL-1beta) in either patients or controls. CONCLUSIONS: The increased concentrations of sICAM-1 in the PF of patients with greater implant scores may indicate an active shedding of the molecule from the endometriotic peritoneal tissue. No significant association was found with the presence of RLs. The levels of IL-1beta in the PF were not a relevant factor influencing the concentrations of sICAM-1. PMID- 12551786 TI - Human chorionic gonadotropin free beta-subunit in the human seminal plasma: a new marker for spermatogenesis? AB - In the past 20 years, several factors were detected in the human seminal plasma and proposed as markers for spermatogenesis. Human chorionic gonadotropin (hCG) and its beta-subunit were therefore investigated, and their seminal levels were found to be higher than those detected in the serum and to correlate with sperm parameters. OBJECTIVE: We designed a retrospective study to determine the suitability of hCG free beta-subunit concentration in the seminal plasma of fertile and infertile male patients as marker of spermatogenesis. STUDY DESIGN: A total of 79 infertile male patients were divided into four groups by their semen analysis results (group 1 [n=8]: azoospermia; group 2 [n=21]: severe oligozoospermia; group 3 [n=40]: oligoasthenospermia (OAS); group 4 [n=10]: asthenospermia) and 10 healthy volunteers of proven fertility were evaluated. RESULTS: The hCG free beta-subunit levels in the seminal plasma were found to be significantly higher (P<0.0001) in the control group in respect to those assayed in the infertile patients and showed a correlation with sperm count (r=0.5) and total motile sperm density (r=0.5). Twenty-five patients were on treatment with oral Mesterolone (100mg daily) plus Tamoxifen (20mg daily) for 3-6 months. Apart from a significant improvement (P<0.05) in sperm morphology, no significant changes in sperm count and motility were observed after the treatment in all the patients. In the seminal plasma of 10 patients who showed a significant increase in sperm count, hCG free beta-subunit levels were found to be significantly higher compared to those detected in the remaining patients (P<0.01). In all patients, these levels remained unchanged after the treatment. CONCLUSIONS: The evidence regarding the positive correlation between hCG free beta-subunit levels in the seminal plasma and sperm concentration is consistent with the previous results regarding hCG levels. A previous study demonstrated that testosterone levels in seminal plasma correlated with sperm concentrations; from the same evidence regarding hCG we hypothesize that seminal plasma testosterone and hCG levels are correlated. Thus, hCG may play a paracrine role in the intratesticular regulation of testosterone secretion. PMID- 12551788 TI - Cat-scratch disease presenting as a solitary tumour in the breast: report of three cases. AB - Cat-scratch disease (CSD) may appear as a solitary mass in the breast and give the impression of a breast carcinoma. In this case, further clinical and laboratory investigation is required to rule out malignancy. We present three cases of CSD of the breast in women of 64, 31 and 61 years old. Each presented with enlarged lymph-nodes in the breast, which were clinically mistaken for solitary tumours. In the first two patients, the mammography was negative. The third patient had a mammogram which indicated a well-defined solid mass in the parenchyma without calcifications. We discuss the clinical, histological and histochemical findings, analyze the differential diagnosis and review the literature. PMID- 12551789 TI - Analysis of retropubic colpourethrosuspension results by suburethral sling with REMEEX prosthesis. AB - OBJECTIVE: To describe the results after colpourethropexy with a REMEEX prosthesis in the treatment of stress urinary incontinence with associated genital prolapse. MATERIALS AND METHODS: A prospective analysis of all patients treated in our Urogynaecology Unit between November 2000 and March 2001. RESULTS: There were 29 patients with mean age of 62 and mean body mass index of 28. All but one patients had stress incontinence (genuine or mixed) associated with a genital prolapse. The other patient was a failed past technique (Marshall-Burch). Cystometry was performed in 80% of the patients. Of these 93% had genuine stress incontinence and 7% mixed incontinence. The average operating time was 119min (CI: 60-310), and the mean hospital stay 5 days (CI: 3-13). There was one case of severe haemorrhage during surgery, four cases of wall seroma and one postoperative urinary infection. Concerning patient satisfaction, 90% of our patients felt they were cured, 8% were much better and 2% had a slight improvement. No patient had a worsening of symptoms after surgery. CONCLUSION: We propose colpourethropexy by suburethral sling with a REMEEX prosthesis as suitable treatment for stress incontinence with associated genital prolapse. PMID- 12551790 TI - Validation of an Italian version of the prolapse quality of life questionnaire. AB - OBJECTIVE: To validate the italian translated version of the prolapse quality of life questionnaire (P-QOL). STUDY DESIGN: The P-QOL questionnaire was translated into Italian and administered to women recruited from a gynaecology outpatient clinic. All women completed a P-QOL questionnaire at the time of the visit, and were examined in left lateral position using the ICS prolapse score. A second P QOL questionnaire was posted and completed by women 1 week later. The validity was assessed by measuring levels of missing data, comparing symptom scores with objective prolapse stages and between affected and asymptomatic women. The reliability was assessed by testing internal consistency and stability using 1 week test retest analysis. RESULTS: 132 symptomatic and 61 asymptomatic women were studied. The number of missing items was 2%. P-QOL domain scores were significantly different between symptomatic and asymptomatic women (P<0.001). Severity according to P-QOL strongly correlated with the prolapse size (P<0.01, rho>0.5). All items achieved a Cronbach alpha greater than 0.8. The test retest reliability confirmed a highly significant correlation between the total scores for each domain. CONCLUSION: The Italian version of the P-QOL questionnaire has been validated, providing a reliable, easily comprehensible and valid instrument for the symptomatic assessment of Italian-speaking women with uterovaginal prolapse. PMID- 12551791 TI - Fluconazole 150 mg single dose versus itraconazole 200 mg per day for 3 days in the treatment of acute vaginal candidiasis: a double-blind randomized study. AB - OBJECTIVE: To compare the safety and efficacy of fluconazole 150mg single dose and itraconazole 200mg per day for 3 days in the treatment of the acute episode of vulvovaginal candidiasis (VVC). METHODS: Double-blind randomized study conducted in three University centers. Patients with acute clinically and mycologically confirmed VVC were enrolled. RESULTS: A total of 86 patients were enrolled; of them, 38 fluconazole and 32 itraconazole patients were evaluable. At the Day 7 visit, all but one fluconazole patients were cured or improved with eradication of the baseline pathogen obtained in all but two itraconazole patients. At the Day 21 visit, a 13% relapse rate was observed in both groups with all other patients cured or improved; eradication rates were 76% for fluconazole and 66% for itraconazole. Global symptom scores (GSS) were significantly more severe at baseline in fluconazole patients (P=0.003). Nevertheless, the slope of the GSS decrease between baseline and Day 7 was similar for both groups whilst GSS were identical at the last visit. Nineteen fluconazole patients reported 31 adverse events and 15 itraconazole patients reported 30 adverse events. CONCLUSIONS: Both oral antifungal treatments showed good clinical and mycological efficacy on the acute episode of VVC with a dramatic decrease of signs and symptoms 7 days after treatment initiation. Fluconazole in single dose warrants optimal compliance in patients who frequently experience more than one episode of VVC. PMID- 12551792 TI - Renal impairment in patients with ovarian cancer. AB - OBJECTIVE: To investigate the prevalence of renal impairment in patients with ovarian cancer at the time of the diagnosis. STUDY DESIGN: Creatinine clearance was estimated according to Cockcroft and Gault (C&G Ccr), glomerular filtration rate (GFR) was determined as renal clearance of 99mTc-DTPA, and renal ultrasound was performed in 60 consecutive patients with newly diagnosed ovarian cancer. RESULTS: A 28% of the total population studied had a GFR <60 ml/min/1.73 m(2). A moderate/severe dilation of the upper urinary tract was found in 12% of patients. The length of kidneys ranged between 9.0 and 13.5 cm, and a statistically significant correlation was found between kidney length and values of GFR. CONCLUSION: A reduction in renal function and a moderate to severe dilation of upper urinary tract frequently occur in patients with ovarian cancer at the time of the diagnosis. PMID- 12551793 TI - Assessment of uterine circulation in ectopic pregnancy by transvaginal color Doppler. AB - OBJECTIVE: The aim of our study was to determine the effect of abnormal implantation on uterine circulation and to evaluate whether the assessment of uterinal blood flow can provide additional information for the diagnosis of tubal pregnancies. METHODS: Forty-nine patients with ectopic pregnancy were examined by transvaginal color Doppler immediately before surgery. Resistance and pulsatility indices of blood flow in the uterine and tubal arteries were measured. RESULTS: The blood flow parameters of the uterine and tubal arteries did not change with gestational age. There was a significant increase in blood flow on the side with the tubal gestation. Differences between sides were higher in the tubal arteries than in the main uterine arteries and showed no dependence on gestational age. CONCLUSION: The abnormal implantation and tubal trophoblast invasion in ectopic pregnancy (EP) can cause more marked blood flow changes in the adjacent supplying vessels than in the main uterine arteries. PMID- 12551794 TI - The effects of pregnancy and lactation on hormonal status and biochemical markers of bone turnover. AB - INTRODUCTION: Biochemical markers of bone turnover are reliable indices for measuring changes in bone formation and bone resorption. Due to limitations in the use of bone densitometry during pregnancy biochemical markers of bone turnover provide an excellent alternative to examine the state of the skeleton during this physiologic state. STUDY DESIGN: We performed a prospective study in 20 women, during their first full term pregnancy until 12 months postpartum, intending to breast feed for 12 (mean, 9.1; range, 7-12) months postpartum. Morning blood and urine samples were obtained for laboratory tests: within 3 months before conception (baseline); between 22 and 24 gestational weeks; after delivery, and 6 and 12 months postpartum. Serum 25-hydroxyvitamin D (25-OH-D), parathyroid hormone (PTH), bone specific alkaline phosphatase, osteocalcin (OC), procollagen I carboxypeptides, calcium, phosphate and creatinine in addition to urine deoxypyridinoline crosslinks and calcium were measured. RESULTS: There was no significant difference in the values of urinary calcium/creatinine and serum calcium, phosphate and 25-OH-D between the different visits during the study. In our patients there was a significant increase in PTH levels at 12 months postpartum as compared to baseline, although the mean values remained in the PTH reference range. All bone turnover markers increased during pregnancy and failed to reach baseline level even 12 months postpartum. CONCLUSION: The high maternal bone turnover may suggest that the calcium needed for infant growth during pregnancy and lactation may be drawn at least in part from the maternal skeleton. PMID- 12551795 TI - Long-term outcome of a randomized study comparing three techniques of conization: cold knife, laser, and LEEP. AB - OBJECTIVE: To evaluate the long-term recurrence rates and complication of different techniques of cervical ablation. METHODS: A randomized trial of three techniques of conization (cold knife, laser, and loop electrosurgical excisional procedure (LEEP)) for cervical intraepithelial neoplasia (CIN) in which 110 patients had been recruited. RESULTS: Eighty-six patients were followed-up for more than 3 years. Of these 28 had been treated with the cold knife, 29 with LEEP and 29 by laser. Five recurrences were observed, one in the cold knife group, two in the LEEP group and two in the laser group (P=NS). The only observed complication was cervical stenosis: zero cases in the laser group, one case in the LEEP group and four cases in the cold knife group (laser versus cold knife: P=0.03; LEEP versus cold knife: P=0.06). Fifty pregnancies were observed in 39 patients. First and second trimester outcomes of pregnancy were without complications. One patient treated with the LEEP presented with a premature rupture of membranes and premature labor at 36 weeks. A total of nine cesarean sections were performed with two cases for cervical dystocia. CONCLUSION: There is no major difference in obstetrical outcome between the three techniques. PMID- 12551796 TI - Recurrence and rapid metastasis formation of a granular cell tumor of the vulva. AB - Histologically confirmed local recurrence of the rare entity of a granular cell tumor of the vulva was diagnosed in a 55-year-old patient with no signs of distant metastasis. Intraoperatively (local excision of mons pubis and inguinal lymphnodes), widespread regional metastasis with retroperitoneal lymphnode metastases were found. Postoperative restaging detected pulmonary, hepatic and skeletal metastases and the patient died of her disseminated disease within 4 months. Early diagnosis of the malignant phenotype of granular cell tumor using expression of S-100-protein, MIB-1, vimentin and p53 is suggested. Once a malignant granular cell tumor is diagnosed histologically, thorough staging procedure should be performed to exclude disseminated disease. If this is the case, radical surgery should be tried due to the lack of efficiency of radiotherapy and of systemic treatments. PMID- 12551797 TI - Pregnancy after hydrothermal endometrial ablation and laparoscopic sterilisation. AB - Hydrothermal ablation (HTA) is a newly introduced technique of endometrial ablation. This report describes the first case of pregnancy and miscarriage following successful HTA performed 4 years after laparoscopic tubal sterilisation. The diagnosis of such pregnancies can be difficult and the risk of pregnancy complications is potentially increased. PMID- 12551798 TI - Fetal arterial redistribution indicating true umbilical cord knot. AB - True umbilical cord knot often remains undiscovered prenatally due to a lack of characteristic clinical or ultrasound signs. We present a pregnancy with favorable outcome in which a non-stress test (NST) found non-reassuring fetal status. Abnormal Doppler blood flow patterns in the descending aorta and in the middle cerebral artery revealed fetal arterial redistribution with normal circulation in the umbilical artery despite a true cord knot. The benefit of fetal Doppler assessment is discussed. Increased aortic-cerebral ratio may reflect acute hypoxic compromise caused by the transitory constriction of the true umbilical cord knot with unrecognized morphologic and circulatory signs. PMID- 12551799 TI - The laparoscopic appearance of Schistosomiasis may be mistaken for "non pigmented" endometriosis. AB - The authors report the case history of a patient with symptoms and signs suggestive of endometriosis, who was found to have Schistosomiasis. The laparoscopic appearance was of gelatinous deposits throughout the pelvis which were thought to be "non-pigmented" endometriosis. However, histological examination of the biopsy specimens revealed Schistosomiasis. PMID- 12551800 TI - A severe case of pubic symphysis diastasis in pregnancy. AB - Pubic symphysis diastasis in pregnancy is a rare occurrence. We report a case of severe pubic symphysis diastasis in pregnancy in a primigravida. PMID- 12551801 TI - Leiomyosarcoma of the breast. A clinicopathologic and immunohistochemical study. AB - Leiomyosarcoma is one of the rarest sarcomas of the breast. We present the clinical, histological and immunohistochemical findings of two new cases of leiomyosarcomas of the breast in 42 and 65 years old women. We analyze the differential diagnostic problems and we review the literature. PMID- 12551802 TI - Second trimester concentration of relaxin and pregnancy related incontinence. PMID- 12551804 TI - Combining anchor and distribution-based methods to derive minimal clinically important differences on the Functional Assessment of Cancer Therapy (FACT) anemia and fatigue scales. AB - Magnitude differences in scores on a measure of quality of life that correspond to differences in function or clinical course are called clinically important differences (CIDs). Anchor-based and distribution-based methods were used to provide ranges of CIDs for five targeted scale scores of the Functional Assessment of Cancer Therapy-Anemia (FACT-An) questionnaire. Three samples of cancer patients were used: Sample 1 included 50 patients participating in a validation study of the FACT-An; Sample 2 included 131 patients participating in a longitudinal study of chemotherapy-induced fatigue; sample 3 included 2,402 patients enrolled in a community-based clinical trial evaluating the effectiveness and safety of a treatment for anemia. Three clinical indicators (hemoglobin level; performance status; response to treatment) were used to determine anchor-based differences. One-half of the standard deviation and 1 standard error of measurement were used as distribution-based criteria. Analyses supported the following whole number estimates of a minimal CID for these five targeted scores: Fatigue Scale = 3.0; FACT-G total score = 4.0; FACT-An total score = 7.0; Trial Outcome Index-Fatigue = 5.0; and Trial Outcome Index-Anemia = 6.0. These estimates provide a basis for sample size estimation when planning for a clinical trial or other longitudinal study, when the purpose is to ensure detection of meaningful change over time. They can also be used in conjunction with more traditional clinical markers to assist investigators in determining treatment efficacy. PMID- 12551805 TI - Relatives' assessment of pain in cognitively impaired nursing home residents. AB - To assess the utility of relatives' assessments of pain in cognitively impaired nursing home residents, the internal consistency of these assessments, and their relationship to other assessments of pain, was examined in a correlational study of 79 residents of a large suburban nursing home. The cohort was 85% female and had an average age of 87 years. The sample included moderately and severely cognitively impaired residents, some of whom were taking pain medication. The results demonstrated that relatives were less likely to rate the pain of the resident when the resident's cognitive level was more impaired and when they had a longer stay in the nursing home. The internal consistency of the relatives' ratings were good and, when only relatives who visited at least once a week were included, correlated significantly with most Minimum Data Set (MDS), resident, physician, and nursing staff ratings. They also correlated significantly with frequency of visits, higher cognitive function, type of relationship with the resident, and with a shorter stay in the nursing home. Relatives' ratings of pain and of past sources of pain may prove useful in the detection of pain in cognitively impaired persons. However, relative's pain ratings are only useful when relatives visit regularly. Like other informants, relatives have more difficulty rating pain when the resident is severely cognitively impaired. PMID- 12551806 TI - Malignant wounds: caregiver-determined clinical problems. AB - To determine the clinical problems common to patients with malignant wounds from the caregivers' perspective, knowledgeable health care providers were asked to list clinical problems from one patient with a malignant wound during malignant wound management workshops. The themes were analyzed using descriptive and exploratory analysis. Data were collected from 136 health care providers. A total of 814 concerns were reported. Patients experienced a mean of 6.00 (95% CI 5.6 6.37) clinical problems each. The problem themes were: physical problems (pain, odor, exudate, bleeding, and edema), emotional stress, functional compromise, social concerns, and complications (e.g., fistulas and nutritional deterioration). This survey of caregivers identified common clinical problems among patients with malignant wounds. It provides the theoretical basis for future quantitative research in populations with malignant wounds. The limitations of this study include observer and recall bias, and it would be beneficial to confirm the findings with patient perspectives by examining patients with variable severity of clinical problems and with multiple tumor types. PMID- 12551807 TI - Phantom pain and risk factors: a multivariate analysis. AB - Phantom pain has been given considerable attention in literature. Phantom pain reduces quality of life, and patients suffering from phantom pain make heavy use of the medical system. Many risk factors have been identified for phantom pain in univariate analyses, including phantom sensations, stump pain, pain prior to the amputation, cause of amputation, prosthesis use, and years elapsed since amputation. Multivariate analyses are lacking in the literature and, therefore, no estimation of an overall risk for phantom pain can be made. The aim of this study was to analyze risk factors in a multivariate analysis in 536 subjects (19% upper limb amputees and 81% lower limb amputees). These subjects filled out a questionnaire in which the following items were assessed; side, date, level, and reason of amputation, pre-amputation pain, presence or absence of phantom pain, phantom sensations and or stump pain, and prosthesis use. The prevalence of phantom pain was 72% (95% CI: 68 to 76%) for the total group, 41% (95% CI: 31 to 51%) in upper limb amputees and 80% (95% CI: 76 to 83%) in lower limb amputees. The most important risk factors for phantom pain were "bilateral amputation" and "lower limb amputation." The risk for phantom pain ranged from 0.33 for a 10-year old patient with a distal upper limb amputation to 0.99 for a subject of 80 years with a bilateral lower limb amputation of which one side is an above knee amputation. PMID- 12551808 TI - Pain prevalence in a French teaching hospital. AB - To measure the prevalence and intensity of pain in hospitalized patients and to assess the quality of pain management, an exhaustive cross-sectional study was conducted in every department in a university hospital. Patients hospitalized for 24 hours or more completed an anonymous self-report questionnaire. Among the 1,475 inpatients, 998 completed the questionnaire. During the 24-hour period prior to our survey, 55% experienced pain. On 100 mm pain intensity measures, the median maximum pain experienced in the 24 preceding hours was 60 mm and the median pain intensity at the time of the survey was 30 mm. Although pain measured at the time of survey disappeared in only 16% of patients, 79% were satisfied with pain management. Despite a high satisfaction level, the prevalence and intensity of pain were very high. This study provided baseline data on pain in a French hospital and led to the implementation of a program for improving pain management. PMID- 12551809 TI - The stability of depression scores in patients who are receiving palliative care. AB - Depression occurs in approximately 25% of palliative care patients. It often is not recognized and is, therefore, not treated. This can lead to difficulties in the management of physical symptoms, such as pain, and also cause much distress to patients and their families. Many professionals working in palliative care are concerned that screening for depression may not be appropriate in a population of patients whose illness is changing rapidly. To explore the value of screening, all patients attending a palliative care day unit were invited to participate in a 12-week study in which they were requested to complete at weekly intervals a previously validated depression-screening tool for palliative care. Fifty patients participated. The scores of all patients who scored below the cut-off threshold at initial assessment showed a mean change of +/-0.56 (range -6 to +7) on the screening tool. This observation suggests that patients' scores on a self completed scale that has been validated for use in the palliative care population remained largely stable during the last few months of life. Screening palliative care patients for depression at referral or first assessment may be useful in assessing depression within the palliative care setting. PMID- 12551810 TI - Pilot study of nasal morphine-chitosan for the relief of breakthrough pain in patients with cancer. AB - Breakthrough pain in patients with cancer is common, often unpredictable, and can rapidly become severe. Treatment using the oral administration of opioids is not optimal due to the slow onset of pain relief. Nasal administration of analgesics potentially offers more rapid pain relief. This study investigates the tolerability and efficacy of a novel morphine-chitosan formulation. Twenty episodes of breakthrough pain were observed in 14 patients with cancer who received 5-80 mg of nasal morphine-chitosan. Nasal symptoms, sedation, giddiness, nausea, and other volunteered symptoms, along with pain scores (pain intensity and pain relief), were recorded at baseline and at regular intervals up to 4 hours after administration, together with an overall satisfaction rating. The formulation was acceptable to patients, generally well tolerated, and had an onset of pain relief 5 minutes after dosing. This formulation warrants further study. PMID- 12551811 TI - Depomedroxyprogesterone acetate for hot flashes. AB - To evaluate the efficacy of a long-acting preparation of medroxyprogesterone acetate for hot flash management, 3 men receiving androgen ablation therapy for prostate cancer and 15 women with a history of breast cancer were treated as part of clinical practice with three biweekly intramuscular injections of 500 mg depomedroxyprogesterone. A review of hot flash diaries and patient charts were completed to evaluate the effectiveness and tolerability of these injections for managing hot flashes. Treatment was associated with an approximate 90% decrease in hot flashes (95% CI 82-97%). Daily hot flash frequency decreased from a mean of 10.9 on the first day of treatment (95% CI 8.0-13.8 hot flashes per day) to a mean of 1.1 hot flashes 6 weeks later (95% CI 0.5-1.8 hot flashes) and to a mean of 0.7 hot flashes 12 weeks following therapy initiation (95% CI 0.1-1.2). Improvement in the hot flashes remained for months after discontinuing the injections in many patients. Reported side effects were minimal. This experience suggests that treatment with depomedroxyprogesterone may be an effective and well tolerated option for the treatment of hot flashes. PMID- 12551812 TI - Symptomatic intravenous antipyretic therapy: efficacy of metamizol, diclofenac, and propacetamol. AB - Fever is a common symptom in cancer patients. The most frequent causes of fever are infections, malignancy itself, various medications, transfusions, and allergy. Although it is necessary to treat the cause of fever, if possible, symptomatic fever management is also important. Surprisingly, little attention is paid to this topic in the medical literature, despite the fact that it is a very frequent problem. In order to support symptomatic fever therapy, we wanted to study the patients' discomfort accompanying fever and the beneficial effects of the symptomatic fever management. To the best of our knowledge, there is an absence of studies in this area, despite the fever discomfort can be an important reason for the antipyretic treatment, mainly in cancer patients. In this non randomized open label pilot study, three intravenous antipyretics were tested in five groups of patients: diclofenac (75 mg, brief intravenous [IV] infusion) vs. metamizol (2500 mg or 1000 mg, brief IV infusion) vs. propacetamol (2000 mg or 1000 mg, slow IV injection or brief IV infusion). The study included 254 febrile episodes mainly in hemato-oncological patients with axillary temperature at least 38 degrees C. The main study endpoints were: changes in axillary temperature, improvement in patient comfort, and number and nature of adverse events. To support justification for symptomatic fever management in febrile patients, we asked the first 45 study subjects to fill in a questionnaire concerning their opinions about fever, fever-associated discomfort, and relief upon antipyretic therapy. All study medications had a significant antipyretic effect. However, metamizol at the dose 2500 mg was considered as the most effective, while propacetamol at the dose 1000 mg showed the lowest antipyretic efficacy. Concerning tolerability and adverse events, there were significant differences among the treatment groups. Diclofenac and metamizol (both 2500 mg and 1000 mg) were tolerated at best. All tested antipyretics significantly improved comfort in febrile patients. Overall, 87% of patients declared improvement in their comfort after administration of antipyretics. Based on the results of the present study, the choice of the antipyretic drug should depend on the clinical status of patient, contraindications, and potential adverse events and risks of the selected agent. It is advisable to use proparacetamol at the higher dosage and to administer it as a brief IV infusion in order to avoid injection-related adverse events. The symptomatic antipyretic treatment in febrile cancer patients is supported by patients themselves and has a significant role in the complex supportive care. Discomfort of patients during fever episodes may be greater than previously thought. PMID- 12551817 TI - Polycythemia vera and the emperor's new clothes. PMID- 12551818 TI - FLT3 inhibition as tailored therapy for acute myeloid leukemia. PMID- 12551819 TI - Genetic abnormalities in marginal zone B-cell lymphoma. PMID- 12551821 TI - Polycythemia vera in young patients: a study on the long-term risk of thrombosis, myelofibrosis and leukemia. AB - BACKGROUND AND OBJECTIVES: Polycythemia vera (PV) is unusual in young patients, so that little information is available on long-term clinical evolution in this particular group. The aim of this study was to define the long-term risk of thrombosis, acute leukemia (AL) and myelofibrosis with myeloid metaplasia (MMM) in young PV patients. DESIGN AND METHODS: From 1975 to 2000, 70 PV patients aged less than 50 years were followed for a median time of 14 years (range 2-26). About three quarters were treated with pipobroman. The Kaplan-Meier method and Cox regression were used for survival analysis. The standardized mortality ratio (SMR) was calculated using Italian age/sex specific mortality rates. RESULTS: The risk of thrombosis increased during the observation period, reaching a plateau of 14% at 10 years and was markedly higher in individuals with a previous history of thrombosis (p=0.0023). No patient had progression into AL or MMM before the 9th year of follow-up. Subsequently, five patients (7%) developed AL and five (7%) MMM, with a 20-year cumulative risk of 15% and 10%, respectively. Overall survival at 20 years was 62%, with nine patients dying of progression into AL or MMM, four of vascular events, one of lung cancer, and four of non-PV-related causes. The SMR was 5.3, indicating a mortality significantly higher than that of the general population (p<0.000). INTERPRETATION AND CONCLUSIONS: This long-term retrospective cohort study shows that although the median survival of young patients with PV exceeds 23 years, their life expectancy is markedly lower than that of the general population because of disease evolution into AL or MMM. PMID- 12551822 TI - Incidence and prognostic value of FLT3 internal tandem duplication and D835 mutations in acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: Cytogenetics is the most important prognostic factor in acute myeloid leukemia (AML). However, a high proportion of patients show normal or intermediate-risk karyotypes. In these patients, other determinants could help to identify those with a higher risk of relapse. Recently, internal tandem duplications (ITD) and D835 mutations in FLT3 tyrosine kinase receptor have been shown to confer a bad prognosis in AML. DESIGN AND METHODS: We analyzed the incidence of these mutations in a total of 208 patients of different AML subsets and their prognostic relevance in non-promyelocytic de novo AML. RESULTS: FLT3 mutations were detected in 24% of de novo AML, 42% of acute promyelocytic leukemia (APL) and 17% of secondary AML. Four patients showed both ITD and D835 mutations. Ninety-four per cent of the patients with FLT3 alterations were classified into the intermediate-risk group. There was no association between the presence of FLT3 alterations and response to induction while the alterations were associated with a worse disease-free survival and event-free survival in both the overall and intermediate-risk patients. INTERPRETATION AND CONCLUSIONS: Our data confirm that any of the mutations in FLT3 confer a bad prognosis in AML. Because of the high prevalence of these mutations within the intermediate-risk group, their detection could be useful to identify patients with a poor prognosis. PMID- 12551823 TI - Interphase fluorescence in situ hybridization assay for the detection of rearrangements of the EVI-1 locus in chromosome band 3q26 in myeloid malignancies. AB - BACKGROUND AND OBJECTIVES: Rearrangements of the EVI-1 locus in chromosome band 3q26 are associated with a poor prognosis in myeloid malignancies. To aid the diagnosis of such aberrations, and possibly disease monitoring, we established an interphase fluorescence in situ hybridization (FISH) assay for the affected breakpoint region. DESIGN AND METHODS: Several overlapping PAC (P1-derived artificial chromosome) clones centromeric to the EVI-1 gene were labeled with a red fluorescent dye, and PAC clones telomeric to EVI-1 with a green fluorochrome. This dual-color probe was hybridized to cytogenetic preparations of cell lines and patients' samples, which were also investigated for the presence of 3q26 rearrangements by chromosome banding analysis. RESULTS: In nuclei without 3q26 rearrangements, two pairs of co-localized red and green signals were observed, while separation of one red/green signal pair or splitting of one red or one green signal was found when 3q26 aberrations were present. The threshold value for true positivity, as determined on 20 samples from patients with myeloid malignancies without 3q26 rearrangements, was 10.2% for separation of one red/green signal pair, and 1% and 1.3% for splitting of one red or one green signal, respectively. In 17 samples from patients with a 3q26 aberration, the percentage of aberrant cells was significantly above these threshold levels. INTERPRETATION AND CONCLUSIONS: We established an interphase FISH assay that efficiently identifies chromosome breakpoints affecting the EVI-1 locus in 3q26, and represents a useful complement to chromosome banding analysis for the detection of such aberrations. PMID- 12551824 TI - Cytogenetic and molecular delineation of a region of chromosome 3q commonly gained in marginal zone B-cell lymphoma. AB - BACKGROUND AND OBJECTIVES: Whole or partial trisomy 3 represents the most recurrent chromosomal abnormality occurring in marginal zone B-cell lymphoma (MZBCL), a distinct subtype of B-cell non-Hodgkin's lymphoma (NHL). By conventional cytogenetic analysis, unbalanced translocations involving chromosome 3 and leading to a partial trisomy 3q were identified in a series of 14 MZBCL patients. Fluorescent in situ hybridization (FISH) experiments were then performed to characterize the breakpoints further and to delineate the extent of the 3q gained region more accurately. DESIGN AND METHODS: We studied 14 cases of MZBCL combining cytogenetics and FISH techniques using specific probes for the long arm of chromosome 3, including the chromosome 3 a satellite probe, a representative panel of yeast artificial chromosome (YAC) clones mapping the chromosomal 3q region (3q11.2 to 3q23) and the chromosome 3 subtelomeric (3q29) probe. RESULTS: In the 14 cases, additional chromosome 3q material was found to be involved in different unbalanced translocations with chromosomes 1, 6, 7, 8, 11, 13, 14, 15, 17, 19 and 21, leading to a derivative chromosome. None of the chromosomal abnormality juxtaposed the 3q regions with the heavy and/or light k and l immunoglobulin gene loci. Eight different breakpoints distributed between the 3q11.2 and the 3q13.32 regions were identified and a common 3q13.32 3q29 overrepresented region was delineated. INTERPRETATION AND CONCLUSIONS: These results suggest that this critical region may be of importance in the pathogenesis of MZBCL and support the hypothesis that a gene dosage effect rather than a specific gene disruption may be involved in the development of this disease. PMID- 12551825 TI - Von Willebrand factor in Italian centenarians. AB - BACKGROUND AND OBJECTIVES: Subjects with blood type O have lower concentration of von Willebrand factor (VWF) than those with type A, B or AB. Since we recently observed that laboratory signs of marked hypercoagulability are compatible with health and longevity in Italian centenarians, we determined VWF and blood groups in healthy centenarians to see whether levels of this marker of endothelial perturbation were altered and whether its correlation with blood groups was similar to that among the general population. DESIGN AND METHODS: In 74 centenarians and in 110 controls (55<45 years old; 55>45 years old), we studied VWF antigen (VWF:Ag), ristocetin co-factor activity (VWF:Rco), multimeric pattern of VWF and cleaving protease (VWF:CP), and plasmin-antiplasmin complexes (PAP). RESULTS: The levels of VWF:Ag and VWF:Rco in centenarians were significantly higher than in controls without significant difference between blood group O or non-O. Fifty-one percent of centenarians have a reduction of the relative proportion of high molecular weight multimers (HMV); furthermore VWF:CP was lower and PAP significantly higher than in young controls. INTERPRETATION AND CONCLUSIONS: The loss of large VWF multimers in 51% of centenarians could depend on degradation protease(s) in the circulation. VWF, a well-known independent predictor of atherothrombotic disease, was increased in centenarians, independently of the blood group, confirming the previous results of a state of hypercoagulability. The finding that the VWF:CP levels are low when VWF levels are high in centenarians could be a corollary of the previous described paradox of successful aging, adding another marker of increased risk of atherothrombosis to the scenario. PMID- 12551826 TI - A method of analyzing procoagulant activity in monocytes at single cell level. AB - BACKGROUND AND OBJECTIVES: Procoagulant activity (PCA) of monocytes is known to play a pivotal role in a variety of physiologic and pathophysiologic processes, such as disseminated intravascular coagulation, atherosclerosis, arterial and venous thromboembolism, cancer-related hypercoagulability and immunopathologies. Until now, PCA has been studied by clotting assays of a whole cell population or at single cell level by analyzing tissue factor antigen, the protein that initiates PCA but does not always correlate with it. Here, we describe a new simple flow cytometric method that allows the PCA of monocytes to be studied at a single cell level by quantifying the fibrin formed around the cells in suspension. DESIGN AND METHODS: Purified fibrinogen was tagged with FITC and added to a recalcified developer plasma containing suitable amounts of heparin in order to inhibit the expansion of clotting, thus limiting the formation of fibrin to the surface of cells with PCA. With appropiate amounts of heparin, in 10 min, large sea urchin-like cells with fibrin needles around some monocytes were formed and, after fixation, cytofluorimetrically analyzed. RESULTS: Blood mononuclear cells isolated and immediately analyzed showed less than 0.1% sea urchin cells. Adherence alone, lipopolysaccharides or ionomycin stimulated expression of PCA in a dose- and time-dependent relationship: after 30 min, 1-3% of the MNC showed PCA, and after 20 h this reached 5-10%. Density separation of monocytes showed that different stimulators act on different maturation stages. Subjects with diabetes express more monocytes with PCA than normal subjects after 30 min stimulation. INTERPRETATION AND CONCLUSIONS: This method allows PCA analysis of monocytes at single cell level and requires only a low number of cells. The signal produced by the fluorescent fibrin is strong and easily analyzed by flow cytometry. The method is suitable for analyzing blood from patients with different pathologies and many conditions under different stimuli. PMID- 12551827 TI - Interleukin-1 gene cluster polymorphisms and risk of coronary artery disease. AB - BACKGROUND AND OBJECTIVES: The pro-inflammatory cytokine interleukin (IL)-1 has been suggested to play a role in atherosclerosis. Several genetic polymorphisms have been described in the genes of the IL-1 cluster and associations with coronary artery disease (CAD) have been reported, although with contrasting results. DESIGN AND METHODS: The associations of a variable number tandem repeat (86 bp) polymorphism in intron 2 of interleukin-1 receptor antagonist (IL1-RA) and of the 511 C/T polymorphism of IL-1b with the risk of CAD were studied. Three hundred and thirty-five case (CAD+) patients with angiographically documented CAD (stenosis >50% in at least one major coronary artery) were compared with 205 unrelated individuals free of CAD signs at angiogram (CAD- controls). One hundred and two (30.5%) CAD+ patients had single-vessel disease (SVD) and 233 (69.5%) multiple-vessel disease (MVD). RESULTS: There was no statistically significant difference in either genotype distribution or allele frequency of both IL-1 RA and IL-1b 511 C/T polymorphisms between CAD+ cases and CAD- controls. Moreover in multivariate analysis, adjusting for multiple comparisons and confounding factors, no difference was found in IL-1 RA genotype distribution between patients with SVD or MVD. INTERPRETATION AND CONCLUSIONS: Our study does not support the association between IL-1 RA intron 2 VNTR and IL-1b 511 C/T polymorphisms and the risk of CAD in individuals undergoing coronary angiography. PMID- 12551828 TI - Different circumstances of the first venous thromboembolism among younger or older heterozygous carriers of the G20210A polymorphism in the prothrombin gene. AB - BACKGROUND AND OBJECTIVES: The G20210A polymorphism in the prothrombin gene is a common cause of inherited thrombophilia. Scarce information is available about the circumstances of the heralding thrombotic manifestation at different ages. The aim of this study was to determine the risk of spontaneous or secondary venous thromboembolism (VTE) among younger and older carriers of the G20210A prothrombin polymorphism. DESIGN AND METHODS: We performed a case-control study, investigating 650 patients with a first objectively documented deep venous thrombosis of the legs or pulmonary embolism and 703 individuals with no history of vascular disease. In all of them we carried out laboratory screening for antithrombin III, protein C and protein S deficiencies, and for the presence of the factor V Leiden and the G20210A prothrombin polymorphisms. RESULTS: After adjustment for other inherited causes of thrombophilia (deficiency of antithrombin III, protein C or S, factor V Leiden) the overall risk for VTE associated with the prothrombin polymorphism was 3.4 times higher than in the controls (95% CI, 2.0 to 5.8). Stratification according to the age and to the circumstances of the first event revealed an increased risk of spontaneous VTE only among the patients older than 45 years in comparison with age-matched controls (odds ratio 4.4, 95% CI 1.8 to 10.6); among the younger individuals the risk was increased for secondary VTE (odds ratio 4.8, 95% CI, 2.3 to 9.8) but not for spontaneous VTE. INTERPRETATION AND CONCLUSIONS: The clinical penetrance of the thrombotic tendency associated with the G20210A prothrombin polymorphism is more expressed in the presence of a circumstantial risk factor (oral contraceptives, pregnancy, surgery, trauma) and in the presence of older age, which acts as an additional circumstantial risk factor. Accordingly, such situations should not discourage from carrying out laboratory screening. PMID- 12551829 TI - Low-molecular-weight heparin (nadroparin) and very low doses of warfarin in the prevention of upper extremity thrombosis in cancer patients with indwelling long term central venous catheters: a pilot randomized trial. AB - BACKGROUND AND OBJECTIVES: Upper extremity thrombosis is a major complication of central venous catheters implanted for chemotherapy in cancer patients. Vitamin K antagonists and low-molecular-weight heparins have been recommended in this setting, but their relative benefit-to-risk ratios have never been compared. DESIGN AND METHODS: A prospective, randomized, open, parallel-group, multicenter trial was performed comparing the antithrombotic efficacy and safety of warfarin and the low-molecular-weight heparin, nadroparin, in cancer patients who had undergone central venous catheter implantation. Warfarin was given orally at a fixed daily dose of 1 mg and nadroparin was injected subcutaneously at a fixed daily dose of 2,850 IU for 90 days, or until venographically-confirmed thrombosis occurred. The primary efficacy outcome was the occurrence of upper extremity thrombosis confirmed by venography performed 90 days after insertion of the catheter, or earlier if symptoms of thrombosis had appeared. Safety end-points were bleeding and thrombocytopenia. RESULTS: Fifty-nine patients were included in the study. A total of 21 and 24 patients in the nadroparin and warfarin groups, respectively, were evaluable for primary efficacy. Six out of the 21 patients in the nadroparin group (28.6%) and 4 out of the 24 patients in the warfarin group (16.7%) had venographically-documented upper extremity thrombosis at day 90 (p=0.48). Safety was satisfactory and similar with both treatments. INTERPRETATION AND CONCLUSIONS: Warfarin at a fixed, very low dose and nadroparin at a fixed, prophylactic dose had comparable benefit-to-risk ratios in the prevention of thrombosis associated with central venous catheters in cancer patients. PMID- 12551830 TI - Recovery of cord blood hematopoietic progenitors after successive freezing and thawing procedures. AB - BACKGROUND AND OBJECTIVES: Cord blood (CB) is a valuable source of stem cells. Most CB units are still cryopreserved in single bags in the world's CB banks. Thawing a single CB unit, dividing it into two parts, expanding the smaller one and refreezing the other would optimize ex vivo expansion of CB progenitors prior to transplantation: expanded and unexpanded cells could be infused together to accelerate early engraftment. DESIGN AND METHODS: The feasibility of refreezing CB samples was investigated by evaluating the effect of 3 successive cryopreservation procedures in 9 CB units. The number and viability of WBC, BFU E, CFU-GM, CFU-MIX, LTC-IC, and the absolute CD34+ cell count were assessed at time 0 and after each thawing. The percentage of CD34 cells expressing CD38, L selectin, VLA-4, VLA-5, H-CAM, LFA-1 and CXCR4 was also evaluated. RESULTS: After three freezing and thawing procedures, WBC counts decreased, while lymphocytes were unchanged. Viability was 90% of basal values after the first thawing and did not change. BFU-E decreased significantly only after the third thawing. CFU-GM and CFU-MIX did not change significantly, nor did LTC-IC, CD34+ cell counts and CAM and CXCR4 expression on CD34+/ CD38-- cells. INTERPRETATION AND CONCLUSIONS: These data show that two successive freeze-thaw procedures do not significantly affect the clonogenic potential and CAM expression of cord blood progenitors. This information could be exploited to devise new options in ex vivo expansion procedures and quality controls prior to transplantation. PMID- 12551831 TI - Marginal zone-related neoplasms of splenic and nodal origin. AB - BACKGROUND: The marginal zone is an anatomically distinct B-cell compartment of lymphoid tissue with an abundant antigenic influx. Among marginal zone-derived lymphomas the WHO classification listed, in addition to extranodal marginal zone B-cell lymphoma of MALT type, two other marginal zone B-cell neoplasms: splenic marginal zone B-cell lymphoma (+/- villous lymphocytes) and nodal marginal zone B cell lymphoma (+/- monocytoid B cells). These two entities are well characterized histologically, but specific biological markers are lacking. Treatment options are heterogeneous, including a watch-and-wait policy, surgery with or without chemotherapy, purine analogs, and interferon. No prospective studies have been conducted so far. INFORMATION SOURCES: Clinical and pathologic data were reviewed by searches of the published medical literature, including searches in PubMed , important printed publications, and abstracts presented at recent hematology and pathology meetings. STATE OF THE ART: Splenic and nodal marginal zone lymphomas are typical low-grade lymphomas with an indolent course. A subset of patients, however, presents with more aggressive disease and have a shorter survival. Clinical and biological prognostic factors identified in reported series are heterogeneous. The role played by hepatitis C virus (HCV) in marginal zone lymphomas is not fully elucidated, but there is demonstration that eradication of HCV infection in splenic lymphoma with villous lymphocytes causes regression of the lymphoma. The optimal treatment has not yet been identified. Retrospective series, however, show that splenectomy is a good option if symptoms from the presence of spleen enlargement or cytopenias need to be treated. The utility of purine analogs and of anti-CD20 immunotherapy needs to be clarified in prospective trials. PERSPECTIVES: Clinicians and pathologists should co-operate to define stringent diagnostic criteria for these indolent disorders. The optimal therapeutic approach and the role of new treatments need to be assessed in prospective clinical trials. PMID- 12551832 TI - Von Willebrand's disease in the year 2003: towards the complete identification of gene defects for correct diagnosis and treatment. AB - BACKGROUND: Von Willebrand's disease (VWD) is an autosomally inherited bleeding disorder caused by a deficiency or abnormality of von Willebrand factor (VWF). VWF is a multimeric adhesive protein which plays an important role in primary hemostasis by promoting platelet adhesion to the subendothelium at sites of vascular injury and platelet-platelet interactions in high shear-rate conditions. It is also the carrier of factor VIII (FVIII), thus indirectly contributing to the coagulation process. VWD has a prevalence of about 1% in the general population, but the figure for clinically relevant cases is lower (about 100/million inhabitants). Bleeding manifestations are heterogeneous: mucosal bleeding is typical of all VWD cases but hemarthrosis and hematomas may also be present when FVIII levels are low. INFORMATION SOURCES: Most cases appear to have a partial quantitative deficiency of VWF (type 1 VWD) with variable bleeding tendency, whereas qualitative variants (type 2 VWD), due to a dysfunctional VWF, are clinically more homogeneous. Type 3 VWD is rare and the patients have a moderate to severe bleeding diathesis because of the virtual absence of VWF, and a recessive pattern of inheritance. The diagnosis of VWD, especially type I, may be difficult, because the laboratory phenotype is highly heterogeneous and is confounded by the fact that factors outside the VWF gene (e.g., blood group) influence VWF levels. An array of tests is usually required to characterize the VWD types of the disorder and establish the best treatment modality. CONCLUSIONS: The aim of treatment is to correct the dual defect of hemostasis, i.e. abnormal coagulation expressed by low levels of FVIII and abnormal platelet adhesion expressed by the prolonged bleeding time (BT). Desmopressin (DDAVP) is the treatment of choice for type 1 VWD because it corrects the FVIII/VWF levels and the prolonged BT in the majority of cases. In type 3 and in severe forms of type 1 and 2 VWD, DDAVP is not effective and for these patients plasma virally inactivated concentrates containing FVIII and VWF are the mainstay of treatment. These concentrates are clinically effective and safe, although they do not always correct the BT. PMID- 12551833 TI - The prevalence and clinical relevance of p27 and cyclin E immunoreactivity in myelodysplastic syndromes. PMID- 12551834 TI - Incidence of trisomy 8 and 9, deletion of D13S319 and D20S108 loci and BCR/ABL translocation in non-treated essential thrombocythemia patients: an analysis of bone marrow cells using interphase fluorescence in situ hybridization. PMID- 12551835 TI - Contribution of the induction of heme oxygenase-1 to etoposide-induced apoptosis in acute myeloblastic leukemia. PMID- 12551836 TI - Expressions of vascular endothelial growth factor and angiopoietin-2 in myeloma cells. PMID- 12551837 TI - The feasibility of reduced-intensity allogeneic hematopoietic stem cell transplantation from a related donor with HLA one-antigen with or without one allele mismatch. PMID- 12551838 TI - Chimerism-directed adoptive immunotherapy in prevention and treatment of post transplant relapse of leukemia in childhood. PMID- 12551839 TI - Inversion of intron 1 of the factor VIII gene for direct molecular diagnosis of hemophilia A. PMID- 12551840 TI - Visualization of gaseous monoxide reception by soluble guanylate cyclase in the rat retina. AB - Immunohistochemistry using novel monoclonal antibodies (mAbs) allowed us to uncover tissue activities of soluble guanylate cyclase (sGC) fine tuned by NO and CO. Upon NO and CO applications in vitro, purified sGC increased the affinity to mAb3221 by 100- and 10-fold, respectively, but not to mAb28131. Immunohistochemistry for gas-generating enzymes revealed that NO occurred in amacrine, bipolar, and Muller's glia cells (MGCs), whereas CO was derived mostly from heme oxygenase (HO)-2 in MGCs. Basal sGC immunoreactivities in vivo to mAb3221 but not to mAb28131 were enhanced by injecting L-arginine and attenuated by blocking NO synthases, suggesting the ability of the former mAb to sense NO. Comparison of mAb-assisted immunohistochemistry suggested that sGC activities were enhanced by zinc protoporphyrin-IX, an HO inhibitor, and repressed completely by blocking NO. However, suggested roles of CO played in situ varied among different retinal layers. In inner plexiform and inner nuclear layers located in the proximity of the cellular NO sources, CO serves as a simple inhibitor of local sGC, while playing roles in housekeeping sGC activation in external limiting membrane standing far from them. These results suggest that CO generated in MGCs is a diffusible gas mediator regulating sGC in both autocrine and paracrine manners. PMID- 12551841 TI - UV induces VEGF through a TNF-alpha independent pathway. AB - Vascular endothelial growth factor (VEGF) is a potent keratinocyte-derived angiogenic factor. Prior reports suggest that following UV irradiation VEGF in keratinocytes is induced primarily by tumor necrosis factor (TNF)- alpha, a cytokine synthesized and secreted by keratinocytes after UV irradiation. We investigated whether blocking TNF-alpha binding to its receptors would inhibit UV induced VEGF expression and secretion in the keratinocyte-derived line SCC-12F. Irradiation with physiologic UV doses (30 mJ/cm2) substantially induced VEGF mRNA in this cell line, as expected, and mRNA induction was followed by increased VEGF in medium conditioned by UV-irradiated cells. Also as expected, TNF-alpha induced VEGF expression and secretion in a dose-dependent manner. Addition of a hexapeptide (Ac-KWIIVW-NH2), known to block TNF-alpha binding to its receptors, abrogated this TNF-alpha effect on VEGF mRNA induction. However, addition of the peptide to cells immediately after UV irradiation did not substantially affect VEGF mRNA induction or secretion into the medium. Our results suggest that VEGF induction after UV irradiation is mediated by multiple mechanisms and that blocking a single pathway does not affect the response. PMID- 12551842 TI - Impaired insulin secretory capacity in mice lacking a functional vitamin D receptor. AB - It was the aim of this study to further explore the functional role of vitamin D in the endocrine pancreas. By gene targeting, we have recently generated mice in which a lacZ reporter gene is driven by the endogenous vitamin D receptor (VDR) promoter. These mice express a functionally inactive mutant VDR. Pancreatic islets but not exocrine pancreas cells showed strong lacZ reporter gene expression in mutant mice. To rule out possible influences of hypocalcemia on pancreatic endocrine function, a rescue diet enriched with calcium, phosphorus, and lactose was fed to wild-type (WT) and VDR mutant mice. The rescue diet normalized body weight and mineral homeostasis in VDR mutants. In glucose tolerance tests, baseline blood glucose levels were unchanged in fasting VDR mutants. However, blood glucose was elevated after oral or subcutaneous glucose loading, and maximum serum insulin levels were reduced by approximately 60% in VDR mutants vs. WT mice on either diet. In addition, insulin mRNA levels were decreased in VDR mutant mice on both diets, whereas pancreatic beta cell mass, islet architecture, and islet neogenesis were normal. These findings clearly establish a molecular role of the vitamin D-responsive elements in pancreatic insulin synthesis and secretion in vivo. PMID- 12551843 TI - Homocysteine-betaine interactions in a murine model of 5,10 methylenetetrahydrofolate reductase deficiency. AB - Hyperhomocysteinemia, a proposed risk factor for cardiovascular disease, is also observed in other common disorders. The most frequent genetic cause of hyperhomocysteinemia is a mutated methylenetetrahydrofolate reductase (MTHFR), predominantly when folate status is impaired. MTHFR synthesizes a major methyl donor for homocysteine remethylation to methionine. We administered the alternate choline-derived methyl donor, betaine, to wild-type mice and to littermates with mild or severe hyperhomocysteinemia due to hetero- or homozygosity for a disruption of the Mthfr gene. On control diets, plasma homocysteine and liver choline metabolite levels were strongly dependent on the Mthfr genotype. Betaine supplementation decreased homocysteine in all three genotypes, restored liver betaine and phosphocholine pools, and prevented severe steatosis in Mthfr deficient mice. Increasing betaine intake did not further decrease homocysteine. In humans with cardiovascular disease, we found a significant negative correlation between plasma betaine and homocysteine concentrations. Our results emphasize the strong interrelationship between homocysteine, folate, and choline metabolism. Hyperhomocysteinemic Mthfr-compromised mice appear to be much more sensitive to changes of choline/betaine intake than do wild-type animals. Hyperhomocysteinemia, in the range of that associated with folate deficiency or with homozygosity for the 677T MTHFR variant, may be associated with disturbed choline metabolism. PMID- 12551844 TI - Physiological electric fields control the G1/S phase cell cycle checkpoint to inhibit endothelial cell proliferation. AB - Vascular endothelial cell (VEC) proliferation is a key event in angiogenesis and is tightly regulated. Electric potential differences exist around the vascular endothelium and give rise to endogenous electric fields (EFs), whether these EFs influence VEC proliferation is unclear. We exposed cultured VECs to applied EFs of physiological strengths for up to 72 h. EF at 50 or 100 mV/mm did not influence cell proliferation, but at 200 mV/mm, cell density, cell growth rate, and mitosis index decreased significantly. EF-induced reduction in VEC proliferation was not due to increased apoptosis, because caspase apoptosis inhibitor Z-VAD-FMK (20 microM), had no effect on this response. Rather, EF responses were mediated via decreased entry of cells into S phase from G1 phase, as shown by flow cytometry. Western blot showed that EFs decreased G1-specific cyclin E expression and increased cyclin/cyclin-dependent kinase complex inhibitor p27kipl expression. Thus EFs controlled VEC proliferation through induction of cell cycle arrest at G1 by down-regulation of cyclin E expression and up-regulation of p27kipl expression, rather than by promoting apoptosis. If control of the cell cycle by endogenous EFs extends beyond VECs, this would be of widespread biological significance in vivo. PMID- 12551845 TI - Human CD38 interferes with HIV-1 fusion through a sequence homologous to the V3 loop of the viral envelope glycoprotein gp120. AB - CD38 is a progression marker in HIV-1 infection, it displays lateral association with CD4, and down-modulates gp120/CD4 binding. The aim of this study was to elucidate the mechanism behind the interplay between CD4, CD38, and HIV-1. We used mouse cell transfectants expressing human CD4 and either CD38 or other CD4 associated molecules to show that CD38 specifically inhibits gp120/CD4 binding. Human cell transfectants expressing truncated forms of CD38 and bioinformatic analysis were used to map the anti-HIV activity and show that it is concentrated in the membrane-proximal region. This region displayed significant sequence similarity with the V3 loop of the HIV-1 gp120 glycoprotein. In line with this similarity, synthetic soluble peptides derived from this region reproduced the anti-HIV effects of full-length CD38 and inhibited HIV-1 and HIV-2 primary isolates from different subtypes and with different coreceptor use. A multiple branched peptide construct presenting part of the sequence of the V3-like region potently and selectively inhibited HIV-1 replication in the nanomolar range. Conversely, a deletion in the V3-like region abrogated the anti-HIV-1 activity of CD38 and its lateral association with CD4. These findings may provide new insights into the early events of HIV-1 fusion and strategies to intervene. PMID- 12551846 TI - Developmental expression profiles and distinct regional estrogen responsiveness suggest a novel role for the steroid receptor coactivator SRC-1 as discriminative amplifier of estrogen signaling in the rat brain. AB - The regional distribution, developmental profiles, and gonadectomy- and estrogen induced changes in the density of transcripts encoding the steroid receptor coactivator-1 (SRC-1) were examined in the female rat brain by semiquantitative in situ hybridization. The results demonstrate striking differences in the abundance of SRC-1 mRNA in discrete brain regions throughout ontogeny. Whereas transcript densities gradually decreased with age in the cerebral cortex, they peaked prominently during the peripubertal period in the hypothalamic medial preoptic area (MPOA) and ventromedial nucleus (VMN). Gonadectomy and estrogen substitution influenced SRC-1 mRNA levels in sexually mature animals in a region specific fashion. Ovariectomy resulted in a down-regulation of SRC-1 mRNA levels in the VMN, a brain region richly endowed with estrogen receptors and playing a major role in neuroendocrine control of reproductive functions. In contrast, SRC 1 transcript levels were significantly up-regulated after estradiol treatment. Interestingly, SRC-1 expression in the cortex was refractory to alterations of the estrogen milieu. The obtained SRC-1 mRNA expression profiles during development clearly demonstrate brain region specificity and regulation by estrogen, thus it is proposed that SRC-1 amplifies estrogen receptor-dependent transcription in a temporally and spatially coordinated manner and therefore contributes to the functional specialization of brain areas involved in the regulation of reproduction. PMID- 12551847 TI - Cloning of shark RAG2 and characterization of the RAG1/RAG2 gene locus. AB - The recombination-activating genes (RAG) encode a site-specific recombinase that is centrally responsible for the rearrangement of genomic V(D)J exons necessary to form functional immunoglobulin and T-cell receptor genes. To help elucidate the origins of the RAG genes, we have cloned the RAG2 gene from the sandbar shark (Carcharhinus plumbeus) and characterized the entire RAG1/RAG2 gene locus. The shark RAG2 protein consists of 520 amino acids, is approximately 50% identical with RAG2 proteins from other vertebrates, and contains the same three domains identified in mammalian RAG2. Residues critical for RAG2 function are conserved in the shark sequence. In common with other vertebrate species, the shark RAG2 coding region lacks introns and is closely linked in opposite orientation to the RAG1 gene. The intergenic region is 9.4 kb, which is considerably larger than of teleosts (2-3 kb) and is comparable to that of tetrapods. This length is partially explained by the presence of several SINE and LINE fragments. The ancestors of the sharks were apparently the first vertebrates in phylogeny to have RAG genes, and our results confirm that the RAG genes have been highly conserved during evolution both in terms of sequence and gene organization. PMID- 12551848 TI - Intracellular calcium signaling through the cADPR pathway is agonist specific in porcine airway smooth muscle. AB - Cyclic ADP-ribose (cADPR) induces intracellular Ca2+ ([Ca2+]i) release in airway smooth muscle, and the cADPR antagonist, 8-amino-cADPR, abolishes [Ca2+]i oscillations elicited by acetylcholine (ACh), suggesting that cADPR is involved during muscarinic receptor activation. Whether the cADPR signaling pathway is common to agonists acting through different G protein-coupled receptors is not known. Using digital video imaging of Fura2-AM loaded porcine airway smooth muscle cells, we examined the effects of the membrane-permeant cADPR antagonist, 8-bromo-cADPR (8Br-cADPR), on the [Ca2+]i responses to ACh, histamine and endothelin-1 (ET-1). In cells preincubated with 100 microM 8Br-cADPR, the [Ca2+]i responses to ACh and ET-1 were significantly attenuated, whereas responses to histamine were not, suggesting agonist specificity of cADPR signaling. The effects of 8Br-cADPR were concentration dependent. We further examined whether muscarinic receptor subtypes specifically couple to this pathway, because in porcine airway smooth muscle cells, ACh activates both M2 and M3 muscarinic receptors coupled to Gai and Gaq, respectively. Methoctramine, an M2-selective antagonist, attenuated the [Ca2+]i responses to Ach, and there was no further attenuation by 8Br-cADPR. In airway smooth muscle, the CD38/cADPR signaling pathway is involved in [Ca2+]i responses to contractile agonists in an agonist specific manner. PMID- 12551849 TI - Sequence and structure-activity relationship of a scorpion venom toxin with nitrergic activity in rabbit corpus cavernosum. AB - An alpha-toxin responsible for nitric oxide (NO) release in rabbit corpus cavernosum (RbCC) was isolated from Tityus serrulatus venom (TSV). The isolated peptide (molecular mass of 7427.66+/-0.15 Da) was identified as Ts3 after determination of Cys residues, N-terminal amino acid analysis, and proteolytic peptide mapping. Ts3 (30 nM) markedly relaxed the RbCC; this response was blocked by the NO synthesis inhibitor N(omega)-nitro-L-arginine methyl ester (100 microM) and the Na+ channel blocker tetrodotoxin (100 nM). Synthetic peptides based on either Ts3 (P1-16, P17-32, P33-48, P49-64, P9-24, P25-40, P41-56, YGLPDKVPTKT) or Bukatoxin (isolated from Buthus martensi Karsch scorpion venom) sequence (Buka11, Buka11-B, PDKVP, PDSEP) were assayed. These peptides slightly relaxed the RbCC, and such an effect was independent of Na+ channel activation or NO release. Our results indicate that Ts3 exerts nitrergic actions and contributes to the relaxing activity of TSV in RbCC, thus providing a valuable tool to investigate the mechanisms underlying nerve activation in erectile tissues, because NO released from nitrergic fibers plays a key role in the erectile process. Our findings revealed the key importance of the Ts3 structure three-dimensional conformation maintenance for biological activity, because linear peptide sequences neither presented substantial relaxations nor was this effect related to nitrergic activity. PMID- 12551850 TI - Two caspase-mediated apoptotic pathways induced by rotenone toxicity in cortical neuronal cells. AB - Our results presented here suggest that cortical neurons degenerate via two caspase-mediated apoptotic pathways when challenged with 0.5 microM rotenone. Although these two pathways can be attributed to the loss of mitochondrial integrity, the triggers for these pathways are likely due to two separate subsequent events (the release of cytochrome c and the emergence of mitochondrial permeability transition [MPT]). Despite some reports suggesting that the release of cytochrome c is a consequence of MPT, the results of our time course experiments suggest otherwise. In fact, we observed that the release of cytochrome c occurred much earlier than MPT; hence, the former is unlikely to be a consequence of the latter. In addition, we observed that the presence of an MPT inhibitor did not attenuate the activation of caspase-3, and that caspase-1 mediated cell death did not exhibit nuclear condensation and DNA fragmentation. Taken together, these results suggest that there are two routes by which cortical neurons degenerate during ischemic injury or in neurodegenerative diseases. PMID- 12551851 TI - IFN-alpha antagonistic activity of HCV core protein involves induction of suppressor of cytokine signaling-3. AB - Eighty percent of patients newly infected with the hepatitis C virus (HCV) develop chronic infection, suggesting that HCV can develop effective strategies to escape the unspecific and specific immune response of the host. Because SOCS molecules have been recognized to be powerful inhibitors of cytokine signaling via the Jak/STAT pathway, virus-induced expression of these molecules should be an efficient instrument to counteract the cellular response toward interferons (IFNs), an essential part of first line antiviral immune response. This study shows that overexpression of HCV core protein inhibits IFN-alpha-induced tyrosine phosphorylation and activation of STAT1 in hepatic cells. With the use of a STAT1 YFP fusion protein, further evidence is given that HCV core is capable to inhibit nuclear translocation of STAT1. Inhibition of STAT1-tyrosine phosphorylation was accompanied by the induction of SOCS3-mRNA expression, suggesting that the HCV core protein impairs IFN-alpha-induced signal transduction via induction of SOCS3 expression. HCV core protein was competent to partially rescue growth of a genetically engineered influenza A virus lacking its own IFN antagonist. These IFN-antagonistic properties of the HCV core protein may be part of the molecular basis of IFN-alpha unresponsiveness in about one-half of chronically infected HCV patients. PMID- 12551852 TI - The challenge of balancing scientific discovery and translation. PMID- 12551853 TI - Clinical application of C-reactive protein for cardiovascular disease detection and prevention. PMID- 12551854 TI - Coming of age of C-reactive protein: using inflammation markers in cardiology. PMID- 12551855 TI - The fire that burns within: C-reactive protein. PMID- 12551856 TI - Coronary revascularization in women in 2003: sex revisited. PMID- 12551857 TI - Treatment of heart failure through stabilization of the cardiac ryanodine receptor. PMID- 12551858 TI - Two-year angiographic and intravascular ultrasound follow-up after implantation of sirolimus-eluting stents in human coronary arteries. AB - BACKGROUND: The safety and efficacy of sirolimus-eluting stenting have been demonstrated, but the outcome of patients treated with this novel technology beyond the first year remains unknown. We sought to evaluate the angiographic, intravascular ultrasound (IVUS), and clinical outcomes of patients treated with sirolimus-eluting stents 2 years after implantation. METHODS AND RESULTS: This study included 30 patients treated with sirolimus-eluting Bx Velocity stenting (slow release [SR], n=15, and fast release [FR], n=15) in Sao Paulo, Brazil. Twenty-eight patients underwent 2-year angiographic and IVUS follow-up. No deaths occurred during the study period. In-stent late loss was slightly greater in the FR group (0.28+/-0.4 mm) than in the SR group (-0.09+/-0.23 mm, P=0.007). No patient had in-stent restenosis. At 2-year follow-up, only 1 patient (FR group) had a 52% diameter stenosis within the lesion segment, which required repeat revascularization. The target-vessel revascularization rate for the entire cohort was 10% (3/30) at 2 years. All other patients had < or =35% diameter stenosis. Angiographic lumen loss at the stent edges was also minimal (in-lesion late loss was 0.33+/-0.42 mm [FR] and 0.13+/-0.29 mm [SR]). In-stent neointimal hyperplasia volume, as detected by IVUS, remained minimal after 2 years (FR= 9.90+/-9 mm3 and SR=10.35+/-9.3 mm3). CONCLUSIONS: This study demonstrates the safety and efficacy of sirolimus-eluting Bx Velocity stents 2 years after implantation in humans. In stent lumen dimensions remained essentially unchanged at 2-year follow-up in the 2 groups, although angiographic lumen loss was slightly higher in the FR group. Restenosis "catch-up" was not found in our patient population. PMID- 12551859 TI - Should volume standards for cardiovascular surgery focus only on high-risk patients? AB - BACKGROUND: Payers and policy makers are attempting to concentrate selected cardiovascular procedures in high-volume centers. A recent analysis of coronary artery bypass grafting (CABG), however, suggests that volume-based referral initiatives should focus only on high-risk patients. METHODS AND RESULTS: Using the national Medicare database (1994 to 1999), we studied the operative mortality in patients undergoing 4 cardiovascular procedures (CABG, aortic valve replacement, mitral valve replacement, and elective abdominal aortic aneurysm repair). We defined 2 categories of patient risk: high-risk (patients in the highest 25th percentile of predicted risk on the basis of a logistic regression model) and low-risk (patients in the lowest 75th percentile). We then compared operative mortality in patients undergoing surgery at very-high volume hospitals (VHVH, highest 20th percentile of procedure volume) and very-low volume hospitals (VLVH, lowest 20th percentile of procedure volume). Absolute differences in operative mortality between VLVH and VHVH were somewhat larger in high-risk patients. However, volume-related differences in mortality were also significant for low-risk patients undergoing one of the 4 procedures. In relative terms, the effect of hospital volume was similar in both high- and low-risk patients. For high- and low-risk patients, the relative risk (RR) of mortality between VHVH and VLVH were nearly equal for CABG (RR=0.78 for low-risk patients, RR=0.77 for high risk patients), aortic valve replacement (0.73 versus 0.76), mitral valve replacement (0.73 versus 0.74), and abdominal aortic aneurysm repair (0.51 versus 0.54). CONCLUSIONS: Although the merits of volume-based referral initiatives can be debated on many grounds, there seems to be little rationale for restricting these initiatives to high-risk patients. PMID- 12551860 TI - Protective role of uncoupling protein 2 in atherosclerosis. AB - BACKGROUND: Uncoupling protein 2 (UCP2) regulates the production of reactive oxygen species in macrophages. However, its role in atherosclerosis is unknown. METHODS AND RESULTS: Irradiated low-density lipoprotein receptor deficient mice (LDLR-/-) were transplanted with bone marrow from either UCP2 deficient mice (Ucp2-/-) or wild type mice (Ucp2+/+). Mice were fed an atherogenic diet for 7 weeks. Engraftment of bone marrow cells was confirmed by the presence of UCP2 protein expression in spleen cell mitochondria of Ucp2+/+ transplanted mice and its absence in Ucp2-/- transplanted mice. Leukocyte counts and plasma cholesterol levels were comparable in both groups. We found a marked increase in atherosclerotic lesion size in the thoracic aorta of Ucp2-/- transplanted mice compared with control Ucp2+/+ transplanted mice (8.3+/-0.9% versus 4.3+/-0.4%, respectively; P<0.005), as well as in the aortic sinus (150 066+/-12 388 microm2 versus 105 689+/-9 727 microm2, respectively; P<0.05). This was associated with increased nitrotyrosine staining, which suggests enhanced oxidative stress. Analysis of plaque composition revealed a significant increase in macrophage accumulation (P<0.05) and apoptosis (P<0.05), along with a decrease in collagen content (P<0.05), suggesting a potentially more vulnerable phenotype. CONCLUSION: These results suggest a protective role for UCP2 against atherosclerosis. PMID- 12551861 TI - C-reactive protein, the metabolic syndrome, and risk of incident cardiovascular events: an 8-year follow-up of 14 719 initially healthy American women. AB - BACKGROUND: The metabolic syndrome describes a high-risk population having 3 or more of the following clinical characteristics: upper-body obesity, hypertriglyceridemia, low HDL, hypertension, and abnormal glucose. All of these attributes, however, are associated with increased levels of C-reactive protein (CRP). METHODS AND RESULTS: We evaluated interrelationships between CRP, the metabolic syndrome, and incident cardiovascular events among 14 719 apparently healthy women who were followed up for an 8-year period for myocardial infarction, stroke, coronary revascularization, or cardiovascular death; 24% of the cohort had the metabolic syndrome at study entry. At baseline, median CRP levels for those with 0, 1, 2, 3, 4, or 5 characteristics of the metabolic syndrome were 0.68, 1.09, 1.93, 3.01, 3.88, and 5.75 mg/L, respectively (P(trend) <0.0001). Over the 8-year follow-up, cardiovascular event-free survival rates based on CRP levels above or below 3.0 mg/L were similar to survival rates based on having 3 or more characteristics of the metabolic syndrome. At all levels of severity of the metabolic syndrome, however, CRP added prognostic information on subsequent risk. For example, among those with the metabolic syndrome at study entry, age-adjusted incidence rates of future cardiovascular events were 3.4 and 5.9 per 1000 person-years of exposure for those with baseline CRP levels less than or greater than 3.0 mg/L, respectively. Additive effects for CRP were also observed for those with 4 or 5 characteristics of the metabolic syndrome. The use of different definitions of the metabolic syndrome had minimal impact on these findings. CONCLUSIONS: These prospective data suggest that measurement of CRP adds clinically important prognostic information to the metabolic syndrome. PMID- 12551862 TI - C-reactive protein increases plasminogen activator inhibitor-1 expression and activity in human aortic endothelial cells: implications for the metabolic syndrome and atherothrombosis. AB - BACKGROUND: Inflammation plays a pivotal role in atherosclerosis. In addition to being a risk marker for cardiovascular disease, much recent data suggest that C reactive protein (CRP) promotes atherogenesis via effects on monocytes and endothelial cells. The metabolic syndrome is associated with significantly elevated levels of CRP. Plasminogen activator inhibitor-1 (PAI-1), a marker of atherothrombosis, is also elevated in the metabolic syndrome and in diabetes, and endothelial cells are the major source of PAI-1. However, there are no studies examining the effect of CRP on PAI-1 in human aortic endothelial cells (HAECs). METHODS AND RESULTS: Incubation of HAECs with CRP results in a time- and dose dependent increase in secreted PAI-1 antigen, PAI-1 activity, intracellular PAI-1 protein, and PAI-1 mRNA. CRP stabilizes PAI-1 mRNA. Inhibitors of endothelial NO synthase, blocking antibodies to interleukin-6 and an endothelin-1 receptor blocker, fail to attenuate the effect of CRP on PAI-1. CRP additionally increased PAI-1 under hyperglycemic conditions. CONCLUSIONS: This study makes the novel observation that CRP induces PAI-1 expression and activity in HAECs and thus has implications for both the metabolic syndrome and atherothrombosis. PMID- 12551863 TI - Selective COX-2 inhibition improves endothelial function in coronary artery disease. AB - BACKGROUND: There is an ongoing debate as to whether the gastrointestinal safety of COX-2 inhibition compared with nonsteroidal antiinflammatory drugs (NSAIDs) may come at the cost of increased cardiovascular events. In view of the large number of patients at cardiovascular risk requiring chronic analgesic therapy with COX-2 inhibitors for arthritic and other inflammatory conditions, the effects of selective COX-2 inhibition on clinically useful surrogates for cardiovascular disease, particularly endothelial function, need to be determined. METHODS AND RESULTS: Fourteen male patients (mean age, 66+/-3 years) with severe coronary artery disease (average of 2.6 vessels with stenosis >75%) undergoing stable background therapy with aspirin and statins were included. The patients received celecoxib (200 mg BID) or placebo for a duration of 2 weeks in a double blind, placebo-controlled, crossover fashion. After each treatment period, flow mediated dilation of the brachial artery, high-sensitivity C-reactive protein, oxidized LDL, and prostaglandins were measured. Celecoxib significantly improved endothelium-dependent vasodilation compared with placebo (3.3+/-0.4% versus 2.0+/ 0.5%, P=0.026), whereas endothelium-independent vasodilation, as assessed by nitroglycerin, remained unchanged (9.0+/-1.6% versus 9.5+/-1.3%, P=0.75). High sensitivity C-reactive protein was significantly lower after celecoxib (1.3+/-0.4 mg/L) than after placebo (1.8+/-0.5 mg/L, P=0.019), as was oxidized LDL (43.6+/ 2.4 versus 47.6+/-2.6 U/L, P=0.028), whereas prostaglandins did not change. CONCLUSIONS: This is the first study to demonstrate that selective COX-2 inhibition improves endothelium-dependent vasodilation and reduces low-grade chronic inflammation and oxidative stress in coronary artery disease. Thus, selective COX-2 inhibition holds the potential to beneficially impact outcome in patients with cardiovascular disease. PMID- 12551864 TI - Taurine and vitamin C modify monocyte and endothelial dysfunction in young smokers. AB - BACKGROUND: Endothelial dysfunction initiated by monocyte-endothelial interactions has previously been observed in many vasculopathies, including chronic cigarette smoking. Taurine, a semiessential amino acid, and vitamin C, a naturally occurring antioxidant, have previously been shown to have endothelial protective effects when exposed to proinflammatory insults. Therefore, we hypothesized that taurine and vitamin C would restore endothelial function in young smokers by modifying monocyte-endothelial interactions. METHODS AND RESULTS: Endothelial-dependent vasodilatation was assessed in vivo using duplex ultrasonography, and monocyte-endothelial interactions were assessed in vitro using endothelial cell culture (human umbilical vein endothelial cells [HUVECs]) with monocyte-conditioned medium (MCM). Endothelial-dependent vasodilatation was significantly impaired in young smokers compared with nonsmokers. Pretreatment of young smokers for 5 days with 2 g/d vitamin C and, more significantly, with 1.5 g/d taurine attenuated this response. MCM taken from smokers impaired the release of nitric oxide and increased the levels of endothelin-1 release from HUVECs. When HUVECs were cultured with MCM from smokers who had been treated with taurine, the levels of nitric oxide and endothelin-1 returned toward control levels. This was attributed to an upregulation in endothelial nitric oxide synthase expression. CONCLUSIONS: These observations suggest that taurine supplementation has a beneficial impact on macrovascular endothelial function, and an investigation of its effect on altered endothelial function in dyslipidemic states is warranted. PMID- 12551865 TI - Xanthine oxidase inhibition reverses endothelial dysfunction in heavy smokers. AB - BACKGROUND: Cigarette smoking causes endothelial dysfunction, possibly through increased oxidant stress. The enzyme xanthine oxidase produces oxidative free radicals. We tested the hypothesis that xanthine oxidase contributes to endothelial dysfunction in cigarette smokers by administering the inhibitor allopurinol. METHODS AND RESULTS: Fourteen cigarette smokers (31+/-4 pack years) and 14 age- and sex-matched healthy non-smoking control subjects participated in a single-blinded, randomized, 2-phase crossover study. All subjects had no other risk factors for atherosclerosis. Inhibition of xanthine oxidase was achieved by a single oral dose of 600 mg of allopurinol on the day of the study. Stimulated nitric oxide endothelial responses were assessed by forearm blood flow responses to intraarterial administration of acetylcholine and bradykinin 4 to 7 hours later; basal nitric oxide was assessed using the nitric oxide synthase inhibitor NG-monomethyl-L-arginine (L-NMMA); and nitroprusside was used to assess sensitivity to nitric oxide. Dilatation produced by acetylcholine was significantly less in smokers (254+/-57%) than healthy controls (390+/-55%) (P=0.009). Allopurinol reversed endothelial dysfunction in smokers (acetylcholine, 463+/-78%, P=0.001) without affecting responses in non-smokers (401+/-80%). Bradykinin responses were also impaired in smokers (P=0.003), and improved with allopurinol, though not significantly (P=0.06). Responses to nitroprusside and L-NMMA were not significantly different between smokers and controls and were not altered by allopurinol. CONCLUSIONS: Smoking-induced endothelial dysfunction of resistance vessels is rapidly reversed with oral allopurinol. These data suggest that xanthine oxidase contributes importantly to endothelial dysfunction caused by cigarette smoking. PMID- 12551866 TI - Effect of nifedipine and cerivastatin on coronary endothelial function in patients with coronary artery disease: the ENCORE I Study (Evaluation of Nifedipine and Cerivastatin On Recovery of coronary Endothelial function). AB - BACKGROUND: Endothelial dysfunction is an important feature of atherosclerosis. Inhibition of the HMG-CoA pathway and of calcium channels improves endothelial function experimentally and in the forearm circulation. Thus, we investigated the effects of a statin and/or a calcium antagonist on coronary endothelial function in patients with coronary artery disease (CAD). METHODS AND RESULTS: In 343 patients undergoing percutaneous coronary intervention in 29 centers, acetylcholine (10(-6) to 10(-4) mol/L) was infused in a coronary segment without angiographically significant CAD. Changes in coronary diameter were measured by quantitative angiography. Endothelium-independent responses were assessed by intracoronary adenosine (1.2 mg/mL) and nitroglycerin (250 microg). Thereafter, patients were randomized in a double-blind manner to placebo, cerivastatin 0.4 mg/d, nifedipine 30 to 60 mg/d, or their combination. Studies were repeated at 6 months. In the most constricted segment, nifedipine but not cerivastatin reduced vasoconstriction to acetylcholine (18.8% versus placebo 10.0%; P<0.05). Patients not taking ACE inhibitors showed a smaller improvement in the placebo group (6.0%), but nifedipine still had an effect (17.0%; P<0.05 versus placebo). Analysis of all evaluable coronary segments revealed an 11% reduction of acetylcholine-induced vasoconstriction in patients receiving nifedipine and cerivastatin (P<0.05 versus placebo). Cerivastatin lowered LDL cholesterol by 35% (P<0.001). CONCLUSIONS: The ENCORE I trial demonstrates that multicenter studies on coronary endothelial function are feasible. After 6 months' treatment, nifedipine improved coronary endothelial function in the most constricted segment. The combination of nifedipine and cerivastatin tended to improve endothelial function; however, this only reached significance in an analysis of all coronary segments. PMID- 12551867 TI - Acute intravenous L-arginine infusion decreases endothelin-1 levels and improves endothelial function in patients with angina pectoris and normal coronary arteriograms: correlation with asymmetric dimethylarginine levels. AB - BACKGROUND: We tested the hypothesis that asymmetric dimethylarginine (ADMA) levels could be elevated and influence endothelin-1 and nitric oxide release and action in patients with cardiac syndrome X (CSX). In addition, we evaluated whether an intravenous infusion of L-arginine would improve endothelial function in these subjects. METHODS AND RESULTS: Nine patients with CSX and 14 control subjects underwent a continuous infusion of L-arginine (0.125 g/min) or saline for 120 minutes. Sixty minutes after L-arginine or saline infusions, an intravenous insulin bolus (0.1 U/kg) combined with a euglycemic clamp was performed. Basal ADMA and endothelin-1 levels were higher in patients with CSX than in controls. At the end of the first hour of infusion, compared with saline, L-arginine infusion increased basal forearm blood flow, nitrite and nitrate (NOx), and forearm cGMP release and decreased endothelin-1. After insulin bolus, during saline, insulin-induced NOx, endothelin-1, and forearm cGMP release was almost abolished. Conversely, L-arginine restored a physiological profile of all endothelial variables compared with control subjects. In control subjects, compared with saline infusion, L-arginine infusion did not modify any parameter. ADMA levels were positively correlated with basal endothelin-1 levels and negatively correlated with insulin-induced incremental levels of NOx and forearm cGMP release. CONCLUSIONS: Plasma ADMA levels are increased in patients with CSX, and they are correlated with increases in endothelin-1 and reductions in insulin induced increments in plasma NOx and cGMP, effects that are reversed by intravenous L-arginine. These data suggest that increased ADMA levels play a role in the abnormal vascular reactivity that is observed in patients with CSX. PMID- 12551868 TI - Long-term results after the glycoprotein IIb/IIIa inhibitor abciximab in unstable angina: one-year survival in the GUSTO IV-ACS (Global Use of Strategies To Open Occluded Coronary Arteries IV--Acute Coronary Syndrome) Trial. AB - BACKGROUND: This study was designed to investigate long-term effects of the glycoprotein IIb/IIIa inhibitor abciximab in patients with acute coronary syndrome without ST elevation who were not scheduled for coronary intervention. METHODS AND RESULTS: A total of 7800 patients were included with an acute coronary syndrome without ST elevation, documented by either elevated cardiac troponin or transient or persistent ST-segment depression. They were randomized to abciximab bolus and 24-hour infusion, abciximab bolus and 48-hour infusion, or matching placebo. The overall 1-year mortality rate was 8.3% (649 patients). One year mortality was 7.8% in the placebo group and 8.2% in the 24-hour and 9.0% in the 48-hour abciximab infusion group. Compared with placebo, the hazard ratio for the 24-hour infusion of abciximab was 1.1 (95% CI 0.86 to 1.29), and for the 48 hour infusion, it was 1.2 (95% CI 0.95 to 1.41). The lack of benefit of abciximab was observed in every subgroup studied. Patients with negative troponin or elevated C-reactive protein had a higher mortality rate after treatment with abciximab for 48 hours than with placebo: 8.5% versus 5.8% in those with negative troponin (P=0.02), 16.3% versus 12.1% in those with elevated C-reactive protein (P=0.04). CONCLUSIONS: Compared with placebo, abciximab did not provide any survival benefit at 1 year in patients admitted with an acute coronary syndrome with ST depression and/or elevated troponin who were not scheduled to undergo early coronary revascularization. In subgroups of patients, in particular those with low cardiac troponin or elevated C-reactive protein, abciximab was associated with excess mortality. PMID- 12551869 TI - Alcohol consumption and plasma concentration of C-reactive protein. AB - BACKGROUND: Moderate alcohol intake has been associated with lower cardiovascular mortality. However, data evaluating the relationship between C-reactive protein (CRP), a predictor of cardiovascular risk, and alcohol consumption are sparse. METHODS AND RESULTS: The relationship between alcohol consumption and CRP was evaluated in a cross-sectional survey and over time among 1732 men and 1101 women participating in the Pravastatin Inflammation/CRP Evaluation Study. CRP levels were lower in those with moderate alcohol intake versus light or occasional intake: in 5 categories of alcohol intake (no alcohol or <1 drink monthly, 1 to 3 drinks monthly, 1 to 4 drinks weekly, 5 to 7 drinks weekly, and > or =2 drinks daily), median CRP levels were 2.60 mg/L (interquartile range (IQR), 1.20 to 5.30 mg/L), 2.20 mg/L (IQR, 1.00 to 4.40 mg/L), 1.70 mg/L (IQR, 0.80 to 3.80 mg/L), 1.60 mg/L (IQR, 0.80 to 3.30 mg/L), and 1.80 mg/L (IQR, 0.80 to 2.90 mg/L), respectively. This relationship was present among men, women not taking hormone replacement therapy, nonsmokers, and those individuals with and without a history of cardiovascular disease (all P<0.001). In multivariate analysis, the relationship between alcohol consumption and CRP remained significant after controlling for multiple traditional cardiovascular risk factors. Alcohol consumption did not significantly affect the change in CRP or lipid levels associated with statin use. CONCLUSION: Moderate alcohol consumption was associated with lower CRP concentrations than no or occasional alcohol intake, an effect that was independent of alcohol-related effects on lipids. Alcohol may attenuate cardiovascular mortality in part through an anti-inflammatory mechanism. PMID- 12551870 TI - Impact of glucose intolerance and insulin resistance on cardiac structure and function: sex-related differences in the Framingham Heart Study. AB - BACKGROUND: Although insulin resistance has been implicated in the pathogenesis of left ventricular (LV) hypertrophy, previous studies have yielded inconsistent results and are limited by referral bias. METHODS AND RESULTS: We examined the relations between echocardiographic LV measurements and glucose tolerance status in 2623 Framingham Study subjects (1514 women, mean age 53 years) free of myocardial infarction and heart failure. We also evaluated the relations of insulin resistance (homeostasis model, HOMA-IR) and LV and left atrial (LA) measures within the normal and abnormal glucose tolerance categories (the latter included impaired glucose tolerance, impaired fasting glucose, and newly diagnosed diabetes). LV mass (adjusted for age, height, heart rate, and systolic blood pressure) increased across categories of worsening glucose tolerance; the trend was more striking in women (P<0.001) compared with men (P=0.054). In subjects with normal (n=2022) and abnormal glucose tolerance (n=327), covariate adjusted LV mass and LV wall thickness increased across HOMA-IR quartiles in women (P<0.001) but not men. In contrast, covariate-adjusted LA size increased with worsening glucose tolerance and across HOMA-IR quartiles in the normal and abnormal glucose tolerance groups in both sexes. Adjustment for body mass index considerably attenuated the relations of LV/LA measures and HOMA-IR, rendering them statistically nonsignificant in the normal glucose tolerance group. CONCLUSIONS: In our large community-based sample, LV mass and wall thickness increased with worsening glucose intolerance, an effect that was more striking in women compared with men. Insulin resistance was associated with increased LV mass in women alone, but this relation was largely accounted for by obesity. PMID- 12551871 TI - Noninvasive assessment of angiogenesis by ultrasound and microbubbles targeted to alpha(v)-integrins. AB - BACKGROUND: Noninvasive methods for characterizing neovessel formation during angiogenesis are currently lacking. We hypothesized that angiogenesis could be imaged with the use of contrast-enhanced ultrasound (CEU) with microbubbles targeted to alpha(v)-integrins. METHODS AND RESULTS: Microbubbles targeted to alpha(v)-integrins were prepared by conjugating echistatin (MB(E)) or monoclonal antibody against murine alpha(v) (MB(alpha)) to their surface. Control microbubbles (MB(c)) were also prepared. The microvascular behavior of these microbubbles was assessed by intravital microscopy of the cremaster muscle in mice treated for 4 days with sustained-release FGF-2. Microvascular retention was much greater (P<0.01) for MB(E) (11+/-6 mm(-3)) and MB(alpha) (10+/-7 mm(-3)) than that for MB(c) (1+/-1 mm(-3)). Retained MB(E) and MB(alpha) attached directly to the microvascular endothelial surface. Microbubble retention in 4 control mice was minimal. Subcutaneous matrigel plugs enriched with FGF-2 were created in 12 mice and studied 10 days later. Neovessels within the matrigel stained positive for alpha(v)-integrins. CEU demonstrated greater (P<0.01) acoustic intensity for MB(E) (16.0+/-5.9 U) and MB(alpha) (17.0+/-5.5 U) compared with MB(c) (5.8+/-2.6 U). The signal from targeted microbubbles (MB(E) and MB(alpha)) correlated well (r=0.90) with the matrigel blood volume determined by CEU perfusion imaging. CONCLUSIONS: CEU with microbubbles targeted for alpha(v) integrins may provide a noninvasive method for assessing therapeutic angiogenesis. PMID- 12551872 TI - Intramyocardial transplantation of autologous endothelial progenitor cells for therapeutic neovascularization of myocardial ischemia. AB - BACKGROUND: We investigated whether catheter-based, intramyocardial transplantation of autologous endothelial progenitor cells can enhance neovascularization in myocardial ischemia. METHODS AND RESULTS: Myocardial ischemia was induced by placement of an ameroid constrictor around swine left circumflex artery. Four weeks after constrictor placement, CD31+ mononuclear cells (MNCs) were freshly isolated from the peripheral blood of each animal. After overnight incubation of CD31+ MNCs in noncoated plates, nonadhesive cells (NA/CD31+ MNCs) were harvested as the endothelial progenitor cell-enriched fraction. Nonadhesive CD31- cells (NA/CD31- MNCs) were also prepared. Autologous transplantation of 10(7) NA/CD31+ MNCs, 10(7) NA/CD31- MNCs, or PBS was performed with a NOGA mapping injection catheter to target ischemic myocardium. In a parallel study, 10(5) human CD34+ MNCs, 10(5) human CD34- MNCs, or PBS was transplanted into ischemic myocardium of nude rats 10 minutes after ligation of the left anterior descending coronary artery. In the swine study, ischemic area by NOGA mapping, Rentrop grade angiographic collateral development, and echocardiographic left ventricular ejection fraction improved significantly 4 weeks after transplantation of NA/CD31+ MNCs but not after injection of NA/CD31- MNCs or PBS. Capillary density in ischemic myocardium 4 weeks after transplantation was significantly greater in the NA/CD31+ MNC group than the control groups. In the rat study, echocardiographic left ventricular systolic function and capillary density were significantly better preserved in the CD34+ MNC group than in the control groups 4 weeks after myocardial ischemia. CONCLUSIONS: These favorable outcomes encourage future clinical trials of catheter-based, intramyocardial transplantation of autologous CD34+ MNCs in the setting of chronic myocardial ischemia. PMID- 12551873 TI - Localization, macromolecular associations, and function of the small heat shock related protein HSP20 in rat heart. AB - BACKGROUND: The small heat shock proteins HSP20, HSP25, alphaB-crystallin, and myotonic dystrophy kinase binding protein (MKBP) may regulate dynamic changes in the cytoskeleton. For example, the phosphorylation of HSP20 has been associated with relaxation of vascular smooth muscle. This study examined the function of HSP20 in heart muscle. METHODS AND RESULTS: Western blotting identified immunoreactive HSP20, alphaB-crystallin, and MKBP in rat heart homogenates. Subcellular fractionation demonstrated that HSP20, alphaB-crystallin, and MKBP were predominantly in cytosolic fractions. Chromatography with molecular sieving columns revealed that HSP20 and alphaB-crystallin were associated in an aggregate of approximately 200 kDa, and alphaB-crystallin coimmunoprecipitated with HSP20. Immunofluorescence microscopy demonstrated that the pattern of HSP20, alphaB crystallin, and actin staining was predominantly in transverse bands. Treatment with sodium nitroprusside led to increases in the phosphorylation of HSP20, as determined with 2-dimensional immunoblots. Incubation of transiently permeabilized myocytes with phosphopeptide analogues of HSP20 led to an increase in the rate of shortening. The increased shortening rate was associated with an increase in the rate of lengthening and a more rapid decay of the calcium transient. CONCLUSIONS: HSP20 is associated with alphaB-crystallin, possibly at the level of the actin sarcomere. Phosphorylated HSP20 increases myocyte shortening rate through increases in calcium uptake and more rapid lengthening. PMID- 12551874 TI - FKBP12.6-mediated stabilization of calcium-release channel (ryanodine receptor) as a novel therapeutic strategy against heart failure. AB - BACKGROUND: The development of heart failure is tightly correlated with a decrease in the stoichiometric ratio for FKBP12.6 binding to the ryanodine receptor (RyR) in the sarcoplasmic reticulum (SR). We report that a new drug, the 1,4-benzothiazepine derivative JTV519, reverses this pathogenic process. JTV519 is known to have a protective effect against Ca2+ overload-induced myocardial injury. METHODS AND RESULTS: Heart failure was produced by 4 weeks of rapid right ventricular pacing, with or without JTV519; SR were then isolated from dog left ventricular (LV) muscles. First, in JTV519-treated dogs, no signs of heart failure were observed after 4 weeks of chronic right ventricular pacing, LV systolic and diastolic functions were largely preserved, and LV remodeling was prevented. Second, JTV519 acutely inhibited both the FK506-induced Ca2+ leak from RyR in normal SR and the spontaneous Ca2+ leak in failing SR. Third, there was no abnormal Ca2+ leak in SR vesicles isolated from JTV519-treated hearts. Fourth, in JTV519-treated hearts, both the stoichiometry of FKBP12.6 binding to RyR and the amount of RyR-bound FKBP12.6 were restored toward the values seen in normal SR. Fifth, in JTV519-untreated hearts, RyR was PKA-hyperphosphorylated, whereas it was reversed in JTV519-treated hearts, returning the channel phosphorylation toward the levels seen in normal hearts. CONCLUSIONS: During the development of experimental heart failure, JTV519 prevented the amount of RyR-bound FKBP12.6 from decreasing. This in turn reduced the abnormal Ca2+ leak through the RyR, prevented LV remodeling, and led to less severe heart failure. PMID- 12551876 TI - Arterial and cardiac aging: major shareholders in cardiovascular disease enterprises: Part III: cellular and molecular clues to heart and arterial aging. PMID- 12551875 TI - Dominant-negative suppression of HCN channels markedly reduces the native pacemaker current I(f) and undermines spontaneous beating of neonatal cardiomyocytes. AB - BACKGROUND: The pacemaker current I(f) contributes to spontaneous diastolic depolarization of cardiac autonomic cells. In heterologous expression, HCN channels exhibit a hyperpolarization-activated inward current similar to I(f). However, the links between HCN genes and native I(f) are largely inferential, and it remains unknown whether I(f) is essential for cardiac pacing. METHODS AND RESULTS: To clarify this situation, we generated a GYG(402-404)AYA pore mutation of HCN2, which rendered the channel nonfunctional and suppressed wild-type HCN2 in a dominant-negative manner in Chinese hamster ovary cells. In addition, HCN2 AYA suppressed I(HCN4) in a dominant-negative manner when coexpressed with wild type HCN4, indicating that the 2 isoforms HCN2 and HCN4 are able to coassemble to form heteromultimeric complexes. Given that HCN2 and HCN4 are the dominant HCN mRNA transcripts in neonatal rat ventricle, we expressed HCN2-AYA in neonatal cardiocytes using adenoviral gene transfer to test the effect of HCN suppression on native I(f). I(f) density was indeed reduced markedly, from 7.8+/-1.6 pA/pF (n=13) in control cells to 0.3+/-0.2 pA/pF (n=11) in HCN2-AYA-infected cells when measured at -130 mV (P<0.001). To probe the effect of HCN on cardiac pacing, we infected spontaneously beating neonatal monolayers with adenoviral vectors expressing wild-type and mutant HCN channels. Infection with HCN2 and HCN4 accelerated the beating rate significantly, to 230.5+/-8.6 bpm (n=12) and 223.5+/ 12.3 bpm (n=10), respectively, compared with control cultures (83.4+/-4.5 bpm, n=13, P<0.001). Conversely, HCN2-AYA completely undermined spontaneous pacing of neonatal cardiocytes. CONCLUSIONS: HCN channels are the major molecular component of native I(f) and are critical for spontaneous beating of neonatal cardiomyocytes. PMID- 12551877 TI - Images in cardiovascular medicine. Himalayan P-waves in a patient with tricuspid atresia. PMID- 12551878 TI - Markers of inflammation and cardiovascular disease: application to clinical and public health practice: A statement for healthcare professionals from the Centers for Disease Control and Prevention and the American Heart Association. PMID- 12551880 TI - Images in cardiovascular medicine. Thrombus in the internal carotid artery complicating an "unstable" atheromatous plaque. PMID- 12551881 TI - Cardiology patient pages. Coronary artery bypass surgery. PMID- 12551882 TI - Doppler strain rate echocardiography versus magnetic resonance imaging. PMID- 12551883 TI - Intravascular beta-radiation may acutely increase coronary collateral blood flow in patients with coronary artery disease. PMID- 12551884 TI - Simvastatin reduces neointimal thickening after experimental angioplasty. PMID- 12551885 TI - New candidates for promoting coronary revascularization: the elderly. PMID- 12551886 TI - Withdrawal of statins in patients with acute coronary syndromes. PMID- 12551887 TI - The SPARK that burns me up. PMID- 12551888 TI - RYR2 and CASQ2 mutations in patients suffering from catecholaminergic polymorphic ventricular tachycardia. PMID- 12551890 TI - Human airway ecto-adenylate kinase. A mechanism to propagate ATP signaling on airway surfaces. AB - Mechanically induced ATP release from human airway epithelial cells regulates mucociliary clearance through cell surface nucleotide receptors. Ectoenzymes detected on these cells were recently shown to terminate ATP-mediated responses by sequential dephosphorylation of extracellular ATP into ADP, AMP, and adenosine. We now demonstrate that an ecto-adenylate kinase (ecto-AK) contributes to the metabolism of adenine nucleotides on human airway epithelial surfaces by the reversible reaction: ATP + AMP 2ADP. This phosphotransferase exhibited a bilateral distribution on polarized primary cultures of human bronchial epithelial cells with a 4-fold higher activity on the mucosal surface. Ecto-AK presented an absolute requirement for magnesium and adenine-based nucleotides. UMP, GMP, and CMP could not substitute for AMP as gamma-phosphate acceptor, and UDP could not replace ADP. Apparent K(m) and V(max) values were 23 +/- 5 microM and 1.1 +/- 0.1 nmol x min(-1) x cm(-2) for ATP and 43 +/- 6 microM and 0.5 +/- 0.1 nmol x min(-1) x cm(-2) for ADP. Ecto-AK accounted for 20% of [gamma (32)P]ATP dephosphorylation, and the impermeant AK inhibitor, diadenosine pentaphosphate, reduced ADPase activity by more than 70% on both epithelial surfaces. Time course experiments on ATP metabolism demonstrated that ecto-AK significantly prolongs effective ATP and ADP concentrations on airway epithelial surfaces for P2 receptor signaling and reduces by 6-fold adenosine production. Our data suggest a role for this nucleotide entrapment cycle in the propagation of purine-mediated mucociliary clearance on human airway epithelial surfaces. PMID- 12551891 TI - Reversal of senescence in mouse fibroblasts through lentiviral suppression of p53. AB - Senescence is generally defined as an irreversible state of G(1) cell cycle arrest in which cells are refractory to growth factor stimulation. In mouse embryo fibroblasts (MEFs), induction of senescence requires the presence of p19(ARF) and p53, as genetic ablation of either of these genes allows escape from senescence and leads to immortalization. We have developed a lentiviral vector that directs the synthesis of a p53-specific short hairpin transcript, which mediates stable suppression of p53 expression through RNA interference. We show that suppression of p53 expression in senescent MEFs leads to rapid cell cycle re entry, is associated with loss of expression of senescence-associated genes, and leads to immortalization. These data indicate that senescence in MEFs is reversible and demonstrate that both initiation and maintenance of senescence is p53-dependent. PMID- 12551892 TI - Characterization of an archaeal multidrug transporter with a unique amino acid composition. AB - The Smr family of multidrug transporters consists of small membrane proteins that extrude various drugs in exchange with protons rendering cells resistant to these drugs. Smr proteins identified to date have been found only in Eubacteria. In this work we present the cloning and characterization of an Smr protein from the archaeon Halobacterium salinarum, the first Smr in the archaeal kingdom. The protein, named Hsmr, was identified through sequence similarity to the Smr family, and the DNA sequence was cloned into an Escherichia coli expression system. Hsmr is heterologously expressed in a functional form despite the difference in lipid composition of the membrane and the lower salt in the cell and its environment. Cells harboring the Hsmr plasmid transport ethidium bromide in an uncoupler-sensitive process and gain resistance to ethidium bromide and acriflavine. Hsmr binds tetraphenylphosphonium (TPP(+)) with a relatively low affinity (K(D) approximately 200 nm) at low salt concentration that increases (K(D) approximately 40 nm) upon the addition of 2 m of either NaCl or KCl. The Hsmr protein contains many of the signature sequence elements of the Smr family and also a high content of negative residues in the loops, characteristic of extreme halophiles. Strikingly, Hsmr is composed of over 40% valine and alanine residues. These residues are clustered at certain regions of the protein in domains that are not important for activity, as judged from lack of conservation and from previous studies with other Smr proteins. We suggest that this high content of alanine and valine residues is a reflection of a "natural" alanine and valine scanning necessitated by the high GC content of the gene. This phenomenon reveals significant sequence elements in small multidrug transporters. PMID- 12551894 TI - Secretory vesicular transport from the Golgi is altered during ATP-binding cassette protein A1 (ABCA1)-mediated cholesterol efflux. AB - Apolipoprotein AI (apoAI)-mediated cholesterol efflux is a process by which cells export excess cellular cholesterol to apoAI to form high density lipoprotein. ATP binding cassette protein A1 (ABCA1) has recently been identified as the key regulator of this process. The pathways of intracellular cholesterol transport during efflux are largely unknown nor is the molecular mechanism by which ABCA1 governs cholesterol efflux well understood. Here, we report that, in both macrophages and fibroblasts, the secretory vesicular transport changes in response to apoAI-mediated cholesterol efflux. Vesicular transport from the Golgi to the plasma membrane increased 2-fold during efflux. This increase in vesicular transport during efflux was observed in both raft-poor and raft-rich vesicle populations originated from the Golgi. Importantly, enhanced vesicular transport in response to apoAI is absent in Tangier fibroblasts, a cell type with deficient cholesterol efflux due to functional ABCA1 mutations. These findings are consistent with an efflux model whereby cholesterol is transported from the storage site to the plasma membrane via the Golgi. ABCA1 may influence cholesterol efflux in part by enhancing vesicular trafficking from the Golgi to the plasma membrane. PMID- 12551893 TI - Genomic organization and evolution of the CX3CR1/CCR8 chemokine receptor locus. AB - The chemokine receptors CCR8 and CX3CR1 are key players in adaptive immunity and are co-receptors for human immunodeficiency virus. We describe here the genomic organization and evolutionary history of both of these genes. CX3CR1 has three promoters that transcribe three separate exons that are spliced with a fourth exon containing the coding region. CCR8 has two promoters. One promoter produces a transcript of two spliced exons, and the other promoter transcribes an exon containing the coding region and lacks introns. We analyzed these promoters in the context of a luciferase reporter and identified several positive and negative regulatory elements. Identification of the genomic organization of these genes in mouse demonstrates a similar organization for CCR8, but mouse CX3CR1 lacks two of the human promoters and has an additional mouse-specific promoter that transcribes only the exon containing the coding region and therefore resembles the organization of the human and mouse CCR8 genes. We also identify two nontranscribed regions that are highly conserved between human and mouse CX3CR1 containing possible regulatory elements. Examination of the CX3CR1 and CCR8 genes and surrounding genomic regions indicates that these genes are the result of the duplication of an ancestral gene prior to the divergence of teleost fish. We characterize single nucleotide polymorphisms in the promoters of human CCR8 and CX3CR1 and establish linkage relationships between CX3CR1 promoter polymorphisms and two previously described CX3CR1 coding polymorphisms associated with human immunodeficiency virus disease progression and arteriosclerosis susceptibility. PMID- 12551895 TI - Crystal structure of the catalytic domain of the PknB serine/threonine kinase from Mycobacterium tuberculosis. AB - With the advent of the sequencing programs of prokaryotic genomes, many examples of the presence of serine/threonine protein kinases in these organisms have been identified. Moreover, these kinases could be classified as homologues of those belonging to the well characterized superfamily of the eukaryotic serine/threonine and tyrosine kinases. Eleven such kinases were recognized in the genome of Mycobacterium tuberculosis. Here we report the crystal structure of an active form of PknB, one of the four M. tuberculosis kinases that are conserved in the downsized genome of Mycobacterium leprae and are therefore presumed to play an important role in the processes that regulate the complex life cycle of mycobacteria. Our structure confirms again the extraordinary conservation of the protein kinase fold and constitutes a landmark that extends this conservation across the evolutionary distance between high eukaryotes and eubacteria. The structure of PknB, in complex with a nucleotide triphosphate analog, reveals an enzyme in the active state with an unprecedented arrangement of the Gly-rich loop associated with a new conformation of the nucleotide gamma-phosphoryl group. It presents as well a partially disordered activation loop, suggesting an induced fit mode of binding for the so far unknown substrates of this kinase or for some modulating factor(s). PMID- 12551896 TI - Structural basis for dimerization of the Grb10 Src homology 2 domain. Implications for ligand specificity. AB - Grb7, Grb10, and Grb14 are members of a distinct family of adapter proteins that interact with various receptor tyrosine kinases upon receptor activation. Proteins in this family contain several modular signaling domains including a pleckstrin homology (PH) domain, a BPS (between PH and SH2) domain, and a C terminal Src homology 2 (SH2) domain. Although SH2 domains are typically monomeric, we show that the Grb10 SH2 domain and also full-length Grb10 gamma are dimeric in solution under physiologic conditions. The crystal structure of the Grb10 SH2 domain at 1.65-A resolution reveals a non-covalent dimer whose interface comprises residues within and flanking the C-terminal alpha helix, which are conserved in the Grb7/Grb10/Grb14 family but not in other SH2 domains. Val-522 in the BG loop (BG3) and Asp-500 in the EF loop (EF1) are positioned to interfere with the binding of the P+3 residue of a phosphopeptide ligand. These structural features of the Grb10 SH2 domain will favor binding of dimeric, turn containing phosphotyrosine sequences, such as the phosphorylated activation loops in the two beta subunits of the insulin and insulin-like growth factor-1 receptors. Moreover, the structure suggests the mechanism by which the Grb7 SH2 domain binds selectively to pTyr-1139 (pYVNQ) in Her2, which along with Grb7 is co-amplified in human breast cancers. PMID- 12551897 TI - The role of helix 1 aspartates and salt bridges in the stability and conversion of prion protein. AB - A key event in the pathogenesis of transmissible spongiform encephalopathies is the conversion of PrP-sen to PrP-res. Morrissey and Shakhnovich (Morrissey, M. P., and Shakhnovich, E. I. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 11293 11298) proposed that the conversion mechanism involves critical interactions at helix 1 (residues 144-153) and that the helix is stabilized on PrP-sen by intra helix salt bridges between two aspartic acid-arginine ion pairs at positions 144 and 148 and at 147 and 151, respectively. Mutants of the hamster prion protein were constructed by replacing the aspartic acids with either asparagines or alanines to destabilize the proposed helix 1 salt bridges. Thermal and chemical denaturation experiments using circular dichroism spectroscopy indicated the overall structures of the mutants are not substantially destabilized but appear to unfold differently. Cell-free conversion reactions performed using ionic denaturants, detergents, and salts (conditions unfavorable to salt bridge formation) showed no significant differences between conversion efficiencies of mutant and wild type proteins. Using conditions more favorable to salt bridge formation, the mutant proteins converted with up to 4-fold higher efficiency than the wild type protein. Thus, although spectroscopic data indicate the salt bridges do not substantially stabilize PrP-sen, the cell-free conversion data suggest that Asp-144 and Asp-147 and their respective salt bridges stabilize PrP sen from converting to PrP-res. PMID- 12551898 TI - Identification of residues important for ligand binding of thromboxane A2 receptor in the second extracellular loop using the NMR experiment-guided mutagenesis approach. AB - The second extracellular loop (eLP2) of the thromboxane A(2) receptor (TP) had been proposed to be involved in ligand binding. Through two-dimensional (1)H NMR experiments, the overall three-dimensional structure of a constrained synthetic peptide mimicking the eLP2 had been determined by our group (Ruan, K.-H., So, S. P., Wu, J., Li, D., Huang, A., and Kung, J. (2001) Biochemistry 40, 275-280). To further identify the residues involved in ligand binding, a TP receptor antagonist, SQ29,548 was used to interact with the synthetic peptide. High resolution two-dimensional (1)H NMR experiments, NOESY, and TOCSY were performed for the peptide, SQ29,548, and peptide with SQ29,548, respectively. Through completed (1)H NMR assignment and by comparing the different spectra, extra peaks were observed on the NOESY spectrum of the peptide with SQ29,548, which implied the contacts between residues of eLP2 at Val(176), Leu(185), Thr(186), and Leu(187) with SQ29,548 at position H2, H7, and H8. Site-directed mutagenesis was used to confirm the possible ligand-binding sites on native human TP receptor. Each of the four residues was mutated to the residues either in the same group, with different structure or different charged. The mutated receptors were then tested for their ligand binding activity. The receptor with V176L mutant retained binding activity to SQ29,548. All other mutations resulted in decreased or lost binding activity to SQ29,548. These mutagenesis results supported the prediction from NMR experiments in which Val(176), Leu(185), Thr(186), and Leu(187) are the possible residues involved in ligand binding. This information facilitates the understanding of the molecular mechanism of thromboxane A(2) binding to the important receptor and its signal transduction. PMID- 12551899 TI - Palmitoylated peptides from the cysteine-rich domain of SNAP-23 cause membrane fusion depending on peptide length, position of cysteines, and extent of palmitoylation. AB - Synaptosome-associated proteins SNAP-23/25, members of a family of proteins essential for exocytosis, have a highly conserved central cysteine-rich domain that plays an important role in membrane targeting. More than one cysteine in this domain is modified by palmitic acid through a thioester linkage. In an effort to address the biological significance of acylation of this domain, we have generated synthetic peptides corresponding to the cysteine-rich region of SNAP-23 and covalently modified the cysteines with palmitic acid. The interaction of acylated and nonacylated peptides with lipid vesicles and natural membranes has been investigated. Our results indicate that palmitoylation is essential for membrane association. The palmitoylated peptides were able to fuse both model and natural membranes. The extent of fusion depended on the length of the peptides and the number and positions of covalently linked palmitic acids. Peptide mediated fusion was suppressed by lysolipid and involved both outer and inner leaflets of the lipid bilayer, which is characteristic of natural membrane fusion. Our results suggest an important role for the cysteine-rich palmitoylated domain of SNAP-23 in promoting membrane fusion in cells. PMID- 12551900 TI - Sarcomere thin filament regulatory isoforms. Evidence of a dominant effect of slow skeletal troponin I on cardiac contraction. AB - Thin filament proteins tropomyosin (Tm), troponin T (TnT), and troponin I (TnI) form an allosteric regulatory complex that is required for normal cardiac contraction. Multiple isoforms of TnT, Tm, and TnI are differentially expressed in both cardiac development and disease, but concurrent TnI, Tm, and TnT isoform switching has hindered assignment of cellular function to these transitions. We systematically incorporated into the adult sarcomere the embryonic/fetal isoforms of Tm, TnT, and TnI by using gene transfer. In separate experiments, greater than 90% of native TnI and 40-50% of native Tm or TnT were specifically replaced. The Ca(2+) sensitivity of tension development was markedly enhanced by TnI replacement but not by TnT or Tm isoform replacement. Titration of TnI replacement from >90% to <30% revealed a dominant functional effect of slow skeletal TnI to modulate regulation. Over this range of isoform replacement, TnI, but not Tm or TnT embryonic isoforms, influenced calcium regulation of contraction, and this identifies TnI as a potential target to modify contractile performance in normal and diseased myocardium. PMID- 12551901 TI - Ca2+-independent protein kinase Cs mediate heterologous desensitization of leukocyte chemokine receptors by opioid receptors. AB - Heterologous desensitization of chemokine receptors by opioids has been considered to contribute to their immunosuppressive effects. Previous studies show that Met-enkephalin, an endogenous opioid, down-regulates chemotaxis of selected chemokine receptors via phosphorylation. In the present study, we further investigated the molecular mechanism of such cross-regulation. Our data showed that preincubation with Met-enkephalin inhibited both MIP-1 alpha-mediated chemotaxis and Ca(2+) flux of monocytes in a dose-dependent manner. The inhibitory effects were maximal using nanomolar concentrations of activating chemokines, a concentration found in physiological conditions. A decrease both in chemokine receptor affinity and in coupling efficiency between receptors and G protein were observed, which directly contributed to the desensitization effects. However, comparing with chemokines such as MIP-1 alpha and MCP-1, opioids did not elicit a calcium flux, failed to induce MIP-1 alpha receptors internalization, and mediated a less potent heterologous desensitization. We hypothesized that these differences might originate from the involvement of different protein kinase C (PKC) isotypes. In our studies, opioid-mediated down-regulation of MIP-1 alpha receptors could be blocked by the general PKC inhibitor calphostin C, but not by the calcium-dependent classic PKC inhibitor Go6976. Western blotting analysis and immunofluorescent staining further showed that only calcium independent PKCs were activated upon opioid stimulation. Thus, opioids achieve desensitization of chemokine receptors via a unique pathway, involving only calcium-independent PKC isotypes. PMID- 12551902 TI - Signaling between focal adhesion kinase and trio. AB - The Trio guanine nucleotide exchange factor functions in neural development in Caenorhabditis elegans and Drosophila and in the development of neural tissues and skeletal muscle in mouse. The association of Trio with the Lar tyrosine phosphatase led us to study the role of tyrosine phosphorylation in Trio function using focal adhesion kinase (FAK). The Lar-interacting domain of Trio is constitutively tyrosine-phosphorylated when expressed in COS-7 cells and was highly phosphorylated when it was co-transfected with FAK. Co-precipitation studies indicated that Trio binds to the FAK amino-terminal domain and to the FAK kinase domain via its SH3 and kinase domains, respectively. Tyrosine phosphorylated FAK and Trio were present mainly in the detergent-insoluble fraction of cell lysates, and co-expression of Trio and FAK resulted in increased amounts of Trio present in the detergent-insoluble fraction. Immunofluorescence of cells co-transfected with FAK and Trio revealed significant co-localization of the proteins at the cell periphery, indicating that they form a stable complex in vivo. A FAK phosphorylation site, tyrosine residue 2737, was identified in subdomain I of the Trio kinase domain. Additionally, in vitro phosphorylation assays and in vivo co-expression studies indicated that Trio enhances FAK kinase activity. These results suggest Trio may be involved in the regulation of focal adhesion dynamics in addition to effecting changes in the actin cytoskeleton through the activation of Rho family GTPases. PMID- 12551903 TI - A novel nontruncating APOB gene mutation, R463W, causes familial hypobetalipoproteinemia. AB - Familial hypobetalipoproteinemia (FHBL), an autosomal co-dominant disorder, is associated with reduced plasma concentrations (<5th percentile for age and sex) of apolipoprotein (apo) B and beta-migrating lipoproteins. To date, only mutations in APOB encoding prematurely truncated apoB have been found in FHBL. We discovered a novel APOB gene mutation, namely R463W, in an extended Christian Lebanese FHBL kindred. Heterozygotes for R463W had the typical FHBL phenotype, whereas homozygotes had barely detectable apoB-100. The effect of the R463W mutation on apoB secretion was examined using transfected McA-RH7777 cells that expressed one of two recombinant human apoBs, namely B48 and B17. In both cases, the mutant proteins (B48RW and B17RW) were retained within the endoplasmic reticulum and were secreted poorly compared with their wild-type counterparts. Pulse-chase analysis showed that secretion efficiencies of B48RW and B17RW were, respectively, 45 and 40% lower than those of the wild-types. Substitution of Arg(463) with Ala in apoB-17 (B17RA) decreased secretion efficiency by approximately 50%, but substitution with Lys (B17RK) had no effect on secretion, indicating that the positive charge was important. Molecular modeling of apoB predicted that Arg(463) was in close proximity to Glu(756) and Asp(456). Substitution of Glu(756) with Gln (B17EQ) had no effect on secretion, but substitution of Asp(456) with Asn (B17DN) decreased secretion to the same extent as B17RW. In co-transfection experiments, the mutant B17RW showed increased binding to microsomal triglyceride transfer protein as compared with wild-type B17. Thus, the naturally occurring R463W mutant reveals a key local domain governing assembly and secretion of apoB-containing lipoproteins. PMID- 12551904 TI - Identification of a novel set of genes regulated by a unique liver X receptor alpha -mediated transcription mechanism. AB - We have reported previously that liver X receptor-alpha (LXRalpha) can mediate a novel cAMP-dependent increase in renin and c-myc gene transcription by binding as a monomer to a unique regulatory element termed the cAMP-negative response element (CNRE). To determine whether this novel action of LXRalpha has global implications on gene regulation, we employed expression profiling to identify other genes regulated by this unique mechanism. Here we report the existence of a set of known and unknown transcripts regulated in parallel with renin. Querying the Celera Mouse Genome Assembly revealed that a majority of these genes contained the consensus CNRE. We have confirmed the functionality of these CNREs by competition for LXRalpha binding via electrophoretic mobility shift assays (EMSA) and by the use of CNRE decoy molecules documenting the abolishment of the cAMP-mediated gene induction. Taken together, these results demonstrate that the interaction between cAMP-activated LXRalpha and the CNRE enhancer element is responsible for widespread changes in gene expression and identify a set of LXRalpha/cAMP-regulated genes that may have important biological implications. PMID- 12551905 TI - The role of the invariant His-1069 in folding and function of the Wilson's disease protein, the human copper-transporting ATPase ATP7B. AB - The copper-transporting ATPase ATP7B is essential for normal distribution of copper in human cells. Mutations in ATP7B lead to Wilson's disease, a severe disorder with neurological and hepatic manifestations. One of the most common disease mutations, a H1069Q substitution, causes intracellular mislocalization of ATP7B (the Wilson's disease protein, WNDP). His-1069 is located in the nucleotide binding domain of WNDP and is conserved in all copper-transporting ATPases from bacteria to mammals; however, the specific role of this His in the structure and function of WNDP remains unclear. We demonstrate that substitution of His-1069 for Gln, Ala, or Cys does not significantly alter the folding of the WNDP nucleotide-binding domain or the proteolytic resistance of the full-length WNDP. In contrast, the function of WNDP is markedly affected by the mutations. The ability to form an acylphosphate intermediate in the presence of ATP is entirely lost in all three mutants, suggesting that His-1069 is important for ATP dependent phosphorylation. Other steps of the WNDP enzymatic cycle are less dependent on His-1069. The H1069C mutant shows normal phosphorylation in the presence of inorganic phosphate; it binds an ATP analogue, beta,gamma imidoadenosine 5'-triphosphate (AMP-PNP), and copper and undergoes nucleotide dependent conformational transitions similar to those of the wild-type WNDP. Although binding of AMP-PNP is not disrupted by the mutation, the apparent affinity for the nucleotide is decreased by 4-fold. We conclude that His-1069 is responsible for proper orientation of ATP in the catalytic site of WNDP prior to ATP hydrolysis. PMID- 12551906 TI - Members of the evolutionarily conserved PMT family of protein O mannosyltransferases form distinct protein complexes among themselves. AB - Protein O-mannosyltransferases (PMTs) initiate the assembly of O-mannosyl glycans, an essential protein modification. Since PMTs are evolutionarily conserved in fungi but are absent in green plants, the PMT family is a putative target for new antifungal drugs, particularly in fighting the threat of phytopathogenic fungi. The PMT family is phylogenetically classified into PMT1, PMT2, and PMT4 subfamilies, which differ in protein substrate specificity. In the model organism Saccharomyces cerevisiae as well as in many other fungi the PMT family is highly redundant, and only the simultaneous deletion of PMT1/PMT2 and PMT4 subfamily members is lethal. In this study we analyzed the molecular organization of PMT family members in S. cerevisiae. We show that members of the PMT1 subfamily (Pmt1p and Pmt5p) interact in pairs with members of the PMT2 subfamily (Pmt2p and Pmt3p) and that Pmt1p-Pmt2p and Pmt5p-Pmt3p complexes represent the predominant forms. Under certain physiological conditions, however, Pmt1p interacts also with Pmt3p, and Pmt5p with Pmt2p, suggesting a compensatory cooperation that guarantees the maintenance of O-mannosylation. Unlike the PMT1/PMT2 subfamily members, the single member of the PMT4 subfamily (Pmt4p) acts as a homomeric complex. Using mutational analyses we demonstrate that the same conserved protein domains underlie both heteromeric and homomeric interactions, and we identify an invariant arginine residue of transmembrane domain two as essential for the formation and/or stability of PMT complexes in general. Our data suggest that protein-protein interactions between the PMT family members offer a point of attack to shut down overall protein O-mannosylation in fungi. PMID- 12551907 TI - The NudA protein in the gastric pathogen Helicobacter pylori is an ubiquitous and constitutively expressed dinucleoside polyphosphate hydrolase. AB - The gastric pathogen Helicobacter pylori harbors one Nudix hydrolase, NudA, that belongs to the nucleoside polyphosphate hydrolase subgroup. In this work, the enzymatic activity of purified recombinant NudA protein was analyzed on a number of nucleoside polyphosphates. This predicted 18.6-kDa protein preferably hydrolyzes diadenosine tetraphosphate, Ap(4)A at a k(cat) of 0.15 s(-1) and a K(m) of 80 microm, resulting in an asymmetrical cleavage of the molecule into ATP and AMP. To study the biological role of this enzyme in H. pylori, an insertion mutant was constructed. There was a 2-7-fold decrease in survival of the mutant as compared with the wild type after hydrogen peroxide exposure but no difference in survival after heat shock or in spontaneous mutation frequency. Western blot analyses revealed that NudA is constitutively expressed in H. pylori at different growth stages and during stress, which would indicate that this protein has a housekeeping function. Given that H. pylori is a diverse species and that all the H. pylori strains tested in this study harbor the nudA gene and show protein expression, we consider NudA to be an important enzyme in this bacterium. PMID- 12551908 TI - Three-dimensional reconstruction of the Saccharomyces cerevisiae multidrug resistance protein Pdr5p. AB - Pdr5p, the major multidrug exporter in Saccharomyces cerevisiae, is a member of the ATP-binding cassette (ABC) superfamily. Pdr5p shares similar mechanisms of substrate recognition and transport with the human MDR1-Pgp, despite an inverted topology of transmembrane and ATP-binding domains. The hexahistidine-tagged Pdr5p multidrug transporter was highly overexpressed in yeast strains where other ABC genes have been deleted. After solubilization and purification, the 160-kDa recombinant Pdr5p has been reconstituted into a lipid bilayer. Controlled detergent removal from Pdr5p-lipid-detergent micelles allowed the production of peculiar square-shaped particles coexisting with liposomes and proteoliposomes. These particles having 11 nm in side were well suited for single particle analysis by electron microscopy. From such analysis, a computed volume has been determined at 25-A resolution, giving insight into the structural organization of Pdr5p. Comparison with the reported structures of different bacterial ABC transporters was consistent with a dimeric organization of Pdr5p in the square particles. Each monomer was composed of three subregions corresponding to a membrane region of about 50 A in height that joins two well separated protruding stalks of about 40 A in height, ending each one with a cytoplasmic nucleotide binding domain (NBD) lobe of about 50-60 A in diameter. The three-dimensional reconstruction of Pdr5p revealed a close arrangement and a structural asymmetric organization of the two NBDs that appeared oriented perpendicularly within a monomer. The existence of different angular positions of the NBDs, with respect to the stalks, suggest rotational movements during the catalytic cycle. PMID- 12551909 TI - Mapping of synergistic components of weakly interacting protein-protein motifs using arrays of paired peptides. AB - Protein-protein recognition usually involves multiple interactions among different motifs that are scattered over protein surfaces. To identify such weak interactions, we have developed a novel double peptide synthesis (DS) method. This method allows us to map protein-protein interactions that involve two linear dis- continuous components from a polypeptide by the use of spatially addressable synergistic pairs of synthetic peptides. The DS procedure is based on the "SPOT" membrane-bound peptide synthesis technique, but to synthesize a mixture of two peptides, it uses both Fmoc (N-(9-fluorenyl)methoxycarbonyl))-alanine and Alloc alanine at the first cycle. This allows their selective deprotection by either piperidine or tributyltin/palladium treatment, respectively. Using SPOT DS, we confirmed as a proof of principle that Elk-1 Ser(383) phosphorylation by ERK-2 kinase is stimulated by the presence of the Elk-1-docking domain. SPOT DS can also be used to dissect protein-protein motifs that define phosphatase substrate affinity. Using this technique, we identified three new regions in the insulin receptor that stimulate the dephosphorylation of the receptor by protein-tyrosine phosphatase (PTP) 1B and presumably increase the selectivity of PTP for this substrate. These data demonstrate that the SPOT DS technique allows the identification of non-linear weakly interacting protein motifs, which are an important determinant of protein kinase and phosphatase substrate specificity and of protein-protein interactions in general. PMID- 12551910 TI - Combined action of ERK and NF kappa B mediates the protective effect of phorbol ester on Fas-induced apoptosis in Jurkat cells. AB - The mechanisms whereby phorbol esters antagonize Fas-induced apoptosis in Jurkat T cells are poorly defined. In the present study, we report that protection from Fas-induced apoptosis by 12-O-tetradecanoylphorbol 13-acetate (TPA) is dependent on both ERK and NF kappa B activation. First, we showed that two specific mitogen activated protein kinase/ERK kinase-inhibitors, PD98059 and U0126, both counteracted TPA-mediated suppression of Fas-induced apoptosis. Moreover, the dose-dependence of U0126-mediated inhibition of ERK phosphorylation correlated with that of reversion of the anti-apoptotic effect of TPA. Second, we observed an excellent correlation between repression of TPA-induced NF kappa B activation by an irreversible inhibitor of I kappa B alpha phosphorylation, BAY11-7082, and its ability to abrogate TPA-induced suppression of Fas-mediated apoptosis. Furthermore, we located the anti-apoptotic effect of both ERK and NF kappa B to lie upstream of the mitochondrial membrane potential depolarization event. Finally, although each inhibitor at optimal, non-toxic concentration by itself only partly reversed TPA-mediated repression of apoptosis, the combination of U0126 and BAY11-7082 completely abolished the anti-apoptotic effect of TPA. Together these findings suggest that TPA-induced activation of ERK and NF kappa B are parallel events that are both required for maximal inhibition of Fas-induced apoptosis in Jurkat T cells. PMID- 12551911 TI - Novel mechanism of the co-regulation of nuclear transport of SmgGDS and Rac1. AB - The armadillo protein SmgGDS promotes guanine nucleotide exchange by small GTPases containing a C-terminal polybasic region (PBR), such as Rac1 and RhoA. Because the PBR resembles a nuclear localization signal (NLS) sequence, we investigated the nuclear transport of SmgGDS with Rac1 or RhoA. We show that the Rac1 PBR has significant NLS activity when it is fused to green fluorescent protein (GFP) or in the context of full-length Rac1. In contrast, the RhoA PBR has very poor NLS activity when it is fused to GFP or in the context of full length RhoA. The nuclear accumulation of both Rac1 and SmgGDS is enhanced by Rac1 activation and diminished by mutation of the Rac1 PBR. Conversely, SmgGDS nuclear accumulation is diminished by interactions with RhoA. An SmgGDS nuclear export signal sequence that we identified promotes SmgGDS nuclear export. These results suggest that SmgGDS. Rac1 complexes accumulate in the nucleus because the Rac1 PBR has NLS activity and because Rac1 supplies the appropriate GTP-dependent signal. In contrast, SmgGDS.RhoA complexes accumulate in the cytoplasm because the RhoA PBR does not have NLS activity. This model may be applicable to other armadillo proteins in addition to SmgGDS, because we demonstrate that activated Rac1 and RhoA also provide stimulatory and inhibitory signals, respectively, for the nuclear accumulation of p120 catenin. These results indicate that small GTPases with a PBR can regulate the nuclear transport of armadillo proteins. PMID- 12551912 TI - Gene structure and M20T polymorphism of the Schistosoma mansoni Sm14 fatty acid binding protein. Molecular, functioanl, and immunoprotection analysis. AB - The Schistosoma mansoni Sm14 antigen belongs to the fatty acid-binding protein family and is considered a vaccine candidate against at least two parasite worms, Fasciola hepatica and S. mansoni. Here the genomic sequence and the polymorphism of Sm14 have been characterized for the first time. We found that the conserved methionine at position 20 is polymorphic, being exchangeable with threonine (M20T). To evaluate the function of the amino acid residue at this position, we have also constructed the mutant Sm14-A20 besides the two native isoforms (Sm14 M20 and Sm14-T20). The three purified recombinant His(6)-tagged Sm14 proteins (rSm14-M20, rSm14-T20, and rSm14-A20) present a predominant beta-barrel structure as shown by CD spectroscopy. Thermal and urea unfolding studies evidenced a higher structural stability of rSm14-M20 over the other forms (rSm14-M20>rSm14 T20>rSm14-A20). All of the Sm14 proteins were able to bind 11 (dansylamino)undecanoic acid (DAUDA) without substantial difference in the binding affinity. However, rSm14-M20 exhibited a higher affinity for natural fatty acids than the rSm14-T20 and rSm14-A20 proteins as judged by competitive experiments against DAUDA (rSm14-M20>rSm14-T20>rSm14-A20). The rSm14-M20 or rSm14 T20 isoforms but not the rSm14-A20 mutant was able to induce significant protection against S. mansoni cercariae challenge in immunized mice. The level of protection efficacy correlates with the extent of structure stability of the recombinant Sm14 isoforms and mutant. PMID- 12551913 TI - Splicing error in E1alpha pyruvate dehydrogenase mRNA caused by novel intronic mutation responsible for lactic acidosis and mental retardation. AB - An intronic point mutation was identified in the E1alpha PDH gene from a boy with delayed development and lactic acidosis, an X-linked disorder associated with a partial defect in pyruvate dehydrogenase (PDH) activity. Protein analysis demonstrated a corresponding decrease in immunoreactivity of the alpha and beta subunits of the PDH complex. In addition to the normal spliced mRNA product of the E1alpha PDH gene, patient samples contained significant levels of an aberrantly spliced mRNA with the first 45 nucleotides of intron 7 inserted in frame between exons 7 and 8. The genomic DNA analysis found no mutation in the coding regions but revealed a hemizygous intronic G to A substitution 26 nucleotides downstream from the normal exon 7 5'-splice site. Splicing experiments in COS-7 cells demonstrated that this point mutation at intron 7 position 26 is responsible for the aberrant splicing phenotype, which involves a switch from the use of the normal 5'-splice site (intron 7 position 1) to the cryptic 5'-splice site downstream of the mutation (intron 7 position 45). The intronic mutation is unusual in that it generates a consensus binding motif for the splicing factor, SC35, which normally binds to exonic enhancer elements resulting in increased exon inclusion. Thus, the aberrant splicing phenotype is most likely explained by the generation of a de novo splicing enhancer motif, which activates the downstream cryptic 5'-splice site. The mutation documented here is a novel case of intron retention responsible for a human genetic disease. PMID- 12551914 TI - A set of loop-1 and -3 structures in the novel vascular endothelial growth factor (VEGF) family member, VEGF-ENZ-7, is essential for the activation of VEGFR-2 signaling. AB - The vascular endothelial growth factor (VEGF) family plays important roles in angiogenesis and vascular permeability. Novel members of the VEGF family encoded in the Orf virus genome, VEGF-E, function as potent angiogenic factors by specifically binding and activating VEGFR-2 (KDR). VEGF-E is about 45% homologous to VEGF-A at amino acid levels, however, the amino acid residues in VEGF-A crucial for the VEGFR-2-binding are not conserved in VEGF-E. To understand the molecular basis of the biological activity of VEGF-E, we have functionally mapped residues important for interaction of VEGF-E with VEGFR-2 by exchanging the domains between VEGF-E(NZ-7) and PlGF, which binds only to VEGFR-1 (Flt-1). Exchange on the amino- and carboxyl-terminal regions had no suppressive effect on biological activity. However, exchange on either the loop-1 or -3 region of VEGF E(NZ-7) significantly reduced activities. On the other hand, introduction of the loop-1 and -3 of VEGF-E(NZ-7) to placenta growth factor rescued the biological activities. The chimera between VEGF-A and VEGF-E(NZ-7) gave essentially the same results. These findings strongly suggest that a common rule exists for VEGFR-2 ligands (VEGF-E(NZ-7) and VEGF-A) that they build up the binding structure for VEGFR-2 through the appropriate interaction between loop-1 and -3 regions. PMID- 12551915 TI - STAM and Hrs are subunits of a multivalent ubiquitin-binding complex on early endosomes. AB - STAM1 and STAM2, which have been identified as regulators of receptor signaling and trafficking, interact directly with Hrs, which mediates the endocytic sorting of ubiquitinated membrane proteins. The STAM proteins interact with the same coiled-coil domain that is involved in the targeting of Hrs to endosomes. In this work, we show that STAM1 and STAM2, as well as an endocytic regulator protein, Eps15, can be co-immunoprecipitated with Hrs both from membrane and cytosolic fractions and that recombinant Hrs, STAM1/STAM2, and Eps15 form a ternary complex. We find that overexpression of Hrs causes a strong recruitment of STAM2 to endosome membranes. Moreover, STAM2, like Hrs and Eps15, binds ubiquitin, and Hrs, STAM2, and Eps15 colocalize with ubiquitinated proteins in clathrin containing endosomal microdomains. The localization of Hrs, STAM2, Eps15, and clathrin to endosome membranes is controlled by the AAA ATPase mVps4, which has been implicated in multivesicular body formation. Depletion of cellular Hrs by small interfering RNA results in a strongly reduced recruitment of STAM2 to endosome membranes and an impaired degradation of endocytosed epidermal growth factor receptors. We propose that Hrs, Eps15, and STAM proteins function in a multivalent complex that sorts ubiquitinated proteins into the multivesicular body pathway. PMID- 12551916 TI - The islet beta cell-enriched RIPE3b1/Maf transcription factor regulates pdx-1 expression. AB - Pancreatic duodenal homeobox factor-1, PDX-1, is required for pancreas development, islet cell differentiation, and the maintenance of beta cell function. Selective expression in the pancreas appears to be principally regulated by Area II, one of four conserved regulatory sequence domains found within the 5'-flanking region of the pdx-1 gene. Detailed mutagenesis studies have identified potential sites of interaction for both positive- and negative acting factors within the conserved sequence blocks of Area II. The islet beta cell-enriched RIPE3b1 transcription factor, the activator of insulin C1 element driven expression, was shown here to also stimulate Area II by binding to sequence blocks 4 and 5 (termed B4/5). Accordingly, B4/5 DNA-binding protein's molecular mass (i.e. 46 kDa), binding specificity, and islet beta cell-enriched distribution were identical to RIPE3b1. Area II-mediated activation was also unaffected upon replacing B4/5 with the insulin C1/RIPE3b1 binding site. In addition, the chromatin immunoprecipitation assay showed that the Area II region of the endogenous pdx-1 gene was precipitated by an antiserum that recognizes the large Maf protein that comprises the RIPE3b1 transcription factor. These results strongly suggest that RIPE3b1/Maf has an important role in generating and maintaining physiologically functional beta cells. PMID- 12551917 TI - YXXL motifs in SH2-Bbeta are phosphorylated by JAK2, JAK1, and platelet-derived growth factor receptor and are required for membrane ruffling. AB - SH2-Bbeta binds to the activated form of JAK2 and various receptor tyrosine kinases. It is a potent stimulator of JAK2, is required for growth hormone (GH) induced membrane ruffling, and increases mitogenesis stimulated by platelet derived growth factor (PDGF) and insulin-like growth factor I. Its domain structure suggests that SH2-Bbeta may act as an adapter protein to recruit downstream signaling proteins to kinase.SH2-Bbeta complexes. SH2-Bbeta is tyrosyl phosphorylated in response to GH and interferon-gamma, stimulators of JAK2, as well as in response to PDGF and nerve growth factor. To begin to elucidate the role of tyrosyl phosphorylation in the function of SH2-Bbeta, we used phosphopeptide mapping, mutagenesis, and a phosphotyrosine-specific antibody to identify Tyr-439 and Tyr-494 in SH2-Bbeta as targets of JAK2 both in vitro and in intact cells. SH2-Bbeta lacking Tyr-439 and Tyr-494 inhibits GH-induced membrane ruffling but still activates JAK2. We provide evidence that JAK1, like JAK2, phosphorylates Tyr-439 and Tyr-494 in SH2-Bbeta and that PDGF receptor phosphorylates SH2-Bbeta on Tyr-439. Therefore, phosphorylated Tyr-439 and/or Tyr 494 in SH2-Bbeta may provide a binding site for one or more proteins linking cytokine receptor.JAK2 complexes and/or receptor tyrosine kinases to the actin cytoskeleton. PMID- 12551918 TI - P2X7 mediates superoxide production in primary microglia and is up-regulated in a transgenic mouse model of Alzheimer's disease. AB - Primary rat microglia stimulated with either ATP or 2'- and 3'-O-(4 benzoylbenzoyl)-ATP (BzATP) release copious amounts of superoxide (O(2)(-)*). ATP and BzATP stimulate O(2)(-)* production through purinergic receptors, primarily the P2X(7) receptor. O(2)(-)* is produced through the activation of the NADPH oxidase. Although both p42/44 MAPK and p38 MAPK were activated rapidly in cells stimulated with BzATP, only pharmacological inhibition of p38 MAPK attenuated O(2)(-)* production. Furthermore, an inhibitor of phosphatidylinositol 3-kinase attenuated O(2)(-)* production to a greater extent than an inhibitor of p38 MAPK. Both ATP and BzATP stimulated microglia-induced cortical cell death indicating this pathway may contribute to neurodegeneration. Consistent with this hypothesis, P2X(7) receptor was specifically up-regulated around beta-amyloid plaques in a mouse model of Alzheimer's disease (Tg2576). PMID- 12551919 TI - Differential effects of superoxide dismutase isoform expression on hydroperoxide induced apoptosis in PC-12 cells. AB - The current study examines the contribution of mitochondria-derived reactive oxygen species (ROS) in tert-butyl-hydroperoxide (TBH)-induced apoptotic signaling using clones of undifferentiated pheochromocytoma (PC-12) cells that stably overexpress the human mitochondrial or cytoplasmic forms of superoxide dismutase (SOD) (viz. Mn-SOD or CuZn-SOD, respectively). Exposure of wild type cells to TBH caused an early generation of ROS (30 min) that resulted in cell apoptosis at 24 h. These responses were attenuated with N-acetylcysteine pretreatment; however, N-acetylcysteine was ineffective in cytoprotection when added after TBH-induced ROS formation. Stable overexpression of SOD isoforms caused a 2- and 3.5-fold elevation in CuZn-SOD and Mn-SOD activities in the cytoplasm and mitochondria, respectively, and 3-fold increases in cellular GSH content. Accordingly, the stable overexpression of Mn-SOD attenuated TBH-induced mitochondrial ROS generation and cell apoptosis. Whereas transient Mn-SOD expression similarly prevented PC-12 apoptosis, this was associated with increases in SOD activity but not GSH, indicating that cytoprotection by Mn-SOD overexpression is related to mitochondrial ROS elimination and not due to increases in cellular GSH content per se. Stable or transient CuZn-SOD overexpression exacerbated cell apoptosis in conjunction with accelerated caspase 3 activation, regardless of cell GSH levels. Collectively, our results support a role for mitochondrial ROS in TBH-induced PC-12 apoptosis that is attenuated by Mn-SOD overexpression and is independent of cellular GSH levels per se. PMID- 12551920 TI - Endoproteolysis of beta-secretase (beta-site amyloid precursor protein-cleaving enzyme) within its catalytic domain. A potential mechanism for regulation. AB - Sequential proteolysis of the amyloid precursor protein (APP) by beta- and gamma secretase activities yields the amyloid beta peptide that is widely deposited in the brains of individuals with Alzheimer's disease. The membrane-anchored aspartyl protease beta-site APP-cleaving enzyme (BACE) exhibits all of the characteristics of a beta-secretase and has been shown to cleave APP at its beta site in vitro and in vivo. We found that BACE undergoes cleavage on a surface exposed alpha-helix between amino acid residues Leu-228 and Ala-229, generating stable N- and C-terminal fragments that remain covalently associated via a disulfide bond. The efficiency of BACE endoproteolysis was observed to depend heavily on cell and tissue type. In contrast to brain where holoprotein was predominant, BACE was found primarily as endoproteolyzed fragments in pancreas, liver, and muscle. In addition, we observed a marked up-regulation of BACE endoproteolysis in C2 myoblasts upon differentiation into multinucleated myotubes, a well established model system of muscle tissue specification. As in liver, BACE exists as endoproteolyzed fragments in the hepatic cell line, HepG2. We found that HepG2 cells are capable of generating amyloid beta peptide, suggesting that endoproteolyzed BACE retains measurable beta-secretase activity. We also found that BACE endoproteolysis occurs only after export from the endoplasmic reticulum, is enhanced in the trans-Golgi network, and is sensitive to inhibitors of vesicular acidification. The membrane-bound proteases tumor necrosis factor alpha-converting enzyme and furin were not found to be responsible for this cleavage nor was BACE observed to mediate its own endoproteolysis by an autocatalytic mechanism. Thus, we characterize a specific processing event that may serve to regulate the enzymatic activity of BACE on a post-translational level. PMID- 12551921 TI - Phosphorylation or glutamic acid substitution at protein kinase C sites on cardiac troponin I differentially depress myofilament tension and shortening velocity. AB - There is evidence that multi-site phosphorylation of cardiac troponin I (cTnI) by protein kinase C is important in both long- and short-term regulation of cardiac function. To determine the specific functional effects of these phosphorylation sites (Ser-43, Ser-45, and Thr-144), we measured tension and sliding speed of thin filaments in reconstituted preparations in which endogenous cTnI was replaced with cTnI phosphorylated by protein kinase C-epsilon or mutated to cTnI S43E/S45E/T144E, cTnI-S43E/S45E, or cTnI-T144E. We used detergent-skinned mouse cardiac fiber bundles to measure changes in Ca(2+)-dependence of force. Compared with controls, fibers reconstituted with phosphorylated cTnI, cTnI S43E/S45E/T144E, or cTnI-S43E/S45E were desensitized to Ca(2+), and maximum tension was as much as 27% lower, whereas fibers reconstituted with cTnI-T144E showed no change. In the in vitro motility assay actin filaments regulated by troponin complexes containing phosphorylated cTnI or cTnI-S43E/S45E/T144E showed both a decrease in Ca(2+) sensitivity and maximum sliding speed compared with controls, whereas filaments regulated by cTnI-S43E/S45E showed only decreased maximum sliding speed and filaments regulated by cTnI-T144E demonstrated only desensitization to Ca(2+). Our results demonstrate novel site specificity of effects of PKC phosphorylation on cTnI function and emphasize the complexity of modulation of the actin-myosin interaction by specific changes in the thin filament. PMID- 12551922 TI - Tip60 is a co-repressor for STAT3. AB - Tip60 (Tat-interactive protein, 60 kDa), a cellular protein with intrinsic histone acetyltransferase activity, is involved in DNA damage repair and apoptosis. Recent studies have suggested that Tip60 acts either as a co-activator or a co-repressor to modulate transcription. In this study, we demonstrate that Tip60 represses reporter gene expression when it is fused to the Gal4 DNA binding domain. We also show that Tip60 associates with histone deacetylase 7 (HDAC7) through its N-terminal zinc finger-containing region and that HDAC7 activity is required for the repressive effect of Tip60. Because endogenous Tip60 interacts with STAT3, we hypothesized that Tip60 might complex with STAT3 and HDAC7 and modulate STAT3-mediated trans-activation. Consistent with this hypothesis, the overexpression of Tip60 represses STAT3-driven reporter gene expression, which can be further potentiated by the co-transfection of HDAC7. Furthermore, interleukin-9-induced c-myc expression, which depends on STAT3 activity, is abrogated by exogenous expression of Tip60. This is the first demonstration of which Tip60 represses STAT3 activity in part through the recruitment of HDAC7. PMID- 12551923 TI - Phosphorylation of Raf-1 by p21-activated kinase 1 and Src regulates Raf-1 autoinhibition. AB - Exposure of cells to mitogens or growth factors stimulates Raf-1 activity through a complex mechanism that involves binding to active Ras, phosphorylation on multiple residues, and protein-protein interactions. Recently it was shown that the amino terminus of Raf-1 contains an autoregulatory domain that can inhibit its activity in Xenopus oocytes. In the present work we show that expression of the Raf-1 autoinhibitory domain blocks extracellular signal-regulated kinase 2 activation by the Raf-1 catalytic domain in mammalian cells. We also show that phosphorylation of Raf-1 on serine 338 by PAK1 and tyrosines 340 and 341 by Src relieves autoinhibition and that this occurs through a specific decrease in the binding of the Raf-1 regulatory domain to its catalytic domain. In addition, we demonstrate that phosphorylation of threonine 491 and serine 494, two phosphorylation sites in the catalytic domain that are required for Raf-1 activation, is unlikely to regulate autoinhibition. These results demonstrate that the autoinhibitory domain of Raf-1 is functional in mammalian cells and that its interaction with the Raf-1 catalytic domain is regulated by phosphorylation of serine 338 and tyrosines 340 and 341. PMID- 12551924 TI - Functional roles of loops 3 and 4 in the cyclic nucleotide binding domain of cyclic AMP receptor protein from Escherichia coli. AB - Cyclic AMP is a ubiquitous secondary message that regulates a large variety of functions. The protein structural motif that binds cAMP is highly conserved with the exception of loops 3 and 4, whose structure and length are variable. The cAMP receptor protein of Escherichia coli, CRP, was employed as a model system to elucidate the functional roles of these loops. Based on the sequence differences between CRP and cyclic nucleotide gated channel, three mutants of CRP were constructed: deletion (residues 54-56 in loop 3 were deleted), insertion (loop 4 was lengthened by 5 residues between Glu-78 and Gly-79) and double mutants. The effects of these mutations on the structure and function of CRP were monitored. Results show that the deletion and insertion mutations do not significantly change the secondary structure of CRP, although the tertiary and quaternary structures are perturbed. The functional data indicate that loop 3 modulates the binding affinities of cAMP and DNA. Although the lengthened loop 4 may have some fine-tuning functions, the specific function of the original loop 4 of CRP remains uncertain. The function consequences of mutation in loop 3 of CRP are similar to that of site A and site B in the regulatory subunits of cyclic AMP dependent protein kinases. Thus, the roles played by loop 3 in CRP may represent a more common mechanism employed by cyclic nucleotide binding domain in modulating ligand binding affinity and intramolecular communication. PMID- 12551925 TI - Activation of Raf-1 signaling by protein kinase C through a mechanism involving Raf kinase inhibitory protein. AB - Protein kinase C (PKC) regulates activation of the Raf-1 signaling cascade by growth factors, but the mechanism by which this occurs has not been elucidated. Here we report that one mechanism involves dissociation of Raf kinase inhibitory protein (RKIP) from Raf-1. Classic and atypical but not novel PKC isoforms phosphorylate RKIP at serine 153 (Ser-153). RKIP Ser-153 phosphorylation by PKC either in vitro or in response to 12-O-tetradecanoylphorbol-13-acetate or epidermal growth factor causes release of RKIP from Raf-1, whereas mutant RKIP (S153V or S153E) remains bound. Increased expression of PKC can rescue inhibition of the mitogen-activated protein (MAP) kinase signaling cascade by wild-type but not mutant S153V RKIP. Taken together, these results constitute the first model showing how phosphorylation by PKC relieves a key inhibitor of the Raf/MAP kinase signaling cascade and may represent a general mechanism for the regulation of MAP kinase pathways. PMID- 12551926 TI - Human rhinovirus selectively modulates membranous and soluble forms of its intercellular adhesion molecule-1 (ICAM-1) receptor to promote epithelial cell infectivity. AB - Human rhinoviruses are responsible for many upper respiratory tract infections. 90% of rhinoviruses utilize intercellular adhesion molecule-1 (ICAM-1) as their cellular receptor, which also plays a critical role in recruitment of immune effector cells. Two forms of this receptor exist; membrane-bound (mICAM-1) and soluble ICAM-1 (sICAM-1). The soluble receptor may be produced independently from the membrane-bound form or it may be the product of proteolytic cleavage of mICAM 1. The ratio of airway epithelial cell expression of mICAM-1 to the sICAM-1 form may influence cell infectivity and outcome of rhinovirus infection. We therefore investigated the effect of rhinovirus on expression of both ICAM-1 receptors in normal human bronchial epithelial cells. We observed separate distinct messenger RNA transcripts coding for mICAM-1 and sICAM-1 in these cells, which were modulated by virus. Rhinovirus induced mICAM-1 expression on epithelial cells while simultaneously down-regulating sICAM-1 release, with consequent increase in target cell infectivity. The role of protein tyrosine kinases was investigated as a potential mechanistic pathway. Rhinovirus infection induced rapid phosphorylation of intracellular tyrosine kinase, which may be critical in up regulation of mICAM-1. Elucidation of the underlying molecular mechanisms involved in differential modulation of both ICAM-1 receptors may lead to novel therapeutic strategies. PMID- 12551927 TI - Epoxycyclohexenone inhibits Fas-mediated apoptosis by blocking activation of pro caspase-8 in the death-inducing signaling complex. AB - Death receptors belong to the tumor necrosis factor receptor family. They can induce apoptosis following engagement with specific ligands and are known to play an important role in the regulation of the immune system. Here we report that epoxycyclohexenone (ECH) inhibits apoptosis induced by anti-Fas antibody, Fas ligand (FasL), or tumor necrosis factor-alpha but not by staurosporine, MG-132, C2-ceramide, or UV irradiation. These results suggest that ECH specifically blocks death receptor-mediated apoptosis. Neither the surface expression of Fas nor the Fas-FasL interaction was influenced by ECH. However, ECH did block the activation of pro-caspase-8 in the death-inducing signaling complex, although recruitment of Fas-associating death domain (FADD) and pro-caspase-8 was not affected. ECH inhibited the enzymatic activity of recombinant active caspase-8 at slightly lower concentrations than it did for active caspase-3 and active caspase 9 in vitro. However, in FasL-treated cells, ECH was only able to inhibit the activation of pro-caspase-8, and it had no effect on the already activated caspase-8 at a concentration that is effective at inhibiting Fas-induced apoptosis. ECH directly bound the large subunit of active caspase-8 that contains the active center cysteine and had a relatively higher affinity to pro-caspase-8. Moreover, compared with pro-caspase-3 and pro-caspase-9, pro-caspase-8 was predominantly depleted by biotinylated ECH with avidin beads in the cell lysates, suggesting that ECH preferentially affects pro-caspase-8. Thus, our results suggest that ECH blocks the self-activation of pro-caspase-8 in the death inducing signaling complex and thus selectively inhibits death receptor-mediated apoptosis. PMID- 12551928 TI - Aggregated and monomeric alpha-synuclein bind to the S6' proteasomal protein and inhibit proteasomal function. AB - The accumulation of aggregated alpha-synuclein is thought to contribute to the pathophysiology of Parkinson's disease, but the mechanism of toxicity is poorly understood. Recent studies suggest that aggregated proteins cause toxicity by inhibiting the ubiquitin-dependent proteasomal system. In the present study, we explore how alpha-synuclein interacts with the proteasome. The proteasome exists as a 26 S and a 20 S species. The 26 S proteasome is composed of the 19 S cap and the 20 S core. Aggregated alpha-synuclein strongly inhibited the function of the 26 S proteasome. The IC(50) of aggregated alpha-synuclein for ubiquitin independent 26 S proteasomal activity was 1 nm. Aggregated alpha-synuclein also inhibited 26 S ubiquitin-dependent proteasomal activity at a dose of 500 nm. In contrast, the IC(50) of aggregated alpha-synuclein for 20 S proteasomal activity was > 1 microm. This suggests that aggregated alpha-synuclein selectively interacts with the 19 S cap. Monomeric alpha-synuclein also inhibited proteasomal activity but with lower affinity and less potency. Recombinant monomeric alpha synuclein inhibited the activity of the 20 S proteasomal core with an IC(50) > 10 microm, exhibited no inhibition of 26 S ubiquitin-dependent proteasomal activity at doses up to 5 microm, and exhibited only partial inhibition (50%) of the 26 S ubiquitin-independent proteasomal activity at doses up to 10 mm. Binding studies demonstrate that both aggregated and monomeric alpha-synuclein selectively bind to the proteasomal protein S6', a subunit of the 19 S cap. These studies suggest that proteasomal inhibition by aggregated alpha-synuclein could be mediated by interaction with S6'. PMID- 12551929 TI - An aldose reductase with 20 alpha-hydroxysteroid dehydrogenase activity is most likely the enzyme responsible for the production of prostaglandin f2 alpha in the bovine endometrium. AB - Prostaglandins are important regulators of reproductive function. In particular, prostaglandin F2 alpha (PGF(2 alpha)) is involved in labor and is the functional mediator of luteolysis to initiate a new estrous cycle in many species. These actions have been extensively studied in ruminants, but the enzymes involved are not clearly identified. Our objective was to identify which prostaglandin F synthase is involved and to study its regulation in the endometrium and in endometrial primary cell cultures. The expression of all previously known prostaglandin F synthases (PGFSs), two newly discovered PGFS-like genes, and a 20 alpha-hydroxysteroid dehydrogenase was studied by Northern blot and reverse transcription PCR. These analyses revealed that none of the known PGFS or the PGFS-like genes were significantly expressed in the endometrium. On the other hand, the 20 alpha-hydroxysteroid dehydrogenase gene was strongly expressed in the endometrium at the time of luteolysis. The corresponding recombinant enzyme has a K(m) of 7 microM for PGH(2) and a PGFS activity higher than the lung PGFS. This enzyme has two different activities with the ability to terminate the estrous cycle; it metabolizes progesterone and synthesizes PGF(2 alpha). Taken together, these data point to this newly identified enzyme as the functional endometrial PGFS. PMID- 12551930 TI - Ggamma subunit-selective G protein beta 5 mutant defines regulators of G protein signaling protein binding requirement for nuclear localization. AB - The signal transducing function of Gbeta(5) in brain is unknown. When studied in vitro Gbeta(5) is the only heterotrimeric Gbeta subunit known to interact with both Ggamma subunits and regulators of G protein signaling (RGS) proteins. When tested with Ggamma, Gbeta(5) interacts with other classical components of heterotrimeric G protein signaling pathways such as Galpha and phospholipase C beta. We recently demonstrated nuclear expression of Gbeta(5) in neurons and brain (Zhang, J. H., Barr, V. A., Mo, Y., Rojkova, A. M., Liu, S., and Simonds, W. F. (2001) J. Biol. Chem. 276, 10284-10289). To gain further insight into the mechanism of Gbeta(5) nuclear localization, we generated a Gbeta(5) mutant deficient in its ability to interact with RGS7 while retaining its ability to bind Ggamma, and we compared its properties to the wild-type Gbeta(5). In HEK-293 cells co-transfection of RGS7 but not Ggamma(2) supported expression in the nuclear fraction of transfected wild-type Gbeta(5). In contrast the Ggamma preferring Gbeta(5) mutant was not expressed in the HEK-293 cell nuclear fraction with either co-transfectant. The Ggamma-selective Gbeta(5) mutant was also excluded from the cell nucleus of transfected PC12 cells analyzed by laser confocal microscopy. These results define a requirement for RGS protein binding for Gbeta(5) nuclear expression. PMID- 12551931 TI - The Notch ligands, Delta1 and Jagged2, are substrates for presenilin-dependent "gamma-secretase" cleavage. AB - The evolutionary conserved Notch signaling pathway is involved in cell fate specification and mediated by molecular interactions between the Notch receptors and the Notch ligands, Delta, Serrate, and Jagged. In this report, we demonstrate that like Notch, Delta1 and Jagged2 are subject to presenilin (PS)-dependent, intramembranous "gamma-secretase" processing, resulting in the production of soluble intracellular derivatives. Moreover, and paralleling the observation that expression of familial Alzheimer's disease-linked mutant PS1 compromises production of Notch S3/NICD, we show that the PS-dependent production of Delta1 cytoplasmic derivatives are also reduced in cells expressing mutant PS1. These studies led us to conclude that a similar molecular apparatus is responsible for intramembranous processing of Notch and it's ligands. To assess the potential role of the cytoplasmic derivative on nuclear transcriptional events, we expressed a Delta1-Gal4VP16 chimera and demonstrated marked transcriptional stimulation of a luciferase-based reporter. Our findings offer the proposal that Delta1 and Jagged2 play dual roles as activators of Notch receptor signaling and as receptors that mediate nuclear signaling events via gamma-secretase-generated cytoplasmic domains. PMID- 12551932 TI - Intracellular human immunodeficiency virus Tat expression in astrocytes promotes astrocyte survival but induces potent neurotoxicity at distant sites via axonal transport. AB - The human immunodeficiency virus (HIV)-Tat protein has been implicated in the neuropathogenesis of HIV infection. However, its role in modulating astroglial neuronal relationships is poorly understood. Astrocyte infection with HIV has been associated with rapid progression of dementia. We thus initially transfected astrocytes with HIV proviral DNA and confirmed Tat production in these cells. Subsequently, using stably Tat-producing asytocyte cell lines, we observed that Tat promoted astrocyte survival by causing a prominent antioxidant effect and resistance to cell injury in these cells. Tat was released extracellularly where it could be taken up by other cells. Tat remained functionally active following uptake and caused long terminal repeat (LTR) transactivation in lymphocytic and astrocytic cell lines. Tat released from astrocytes caused mitochondrial dysfunction, trimming of neurites, and cell death in neurons. Tat neurotoxicity was attenuated by anti-Tat antibodies, kynurenate or heparan sulfate. The neurotoxic effects of Tat were caused at concentrations lower than that needed to cause LTR transactivation. When Tat-expressing cells were injected into the rat dentate gyrus, Tat was taken up by granule cells and transported along neuronal pathways to the CA3 region where it caused glial cell activation and neurotoxicity. The arginine-rich domain of Tat was essential for both the LTR transactivation and the neurotoxic properties of Tat. Thus HIV-Tat is a potent neurotoxin that may act at distant sites while at the same time it assures its production by preventing cell death in astrocytes where it is produced. PMID- 12551933 TI - Synthesis and functional analyses of nuclear clusterin, a cell death protein. AB - Nuclear clusterin (nCLU) is an ionizing radiation (IR)-inducible protein that binds Ku70, and triggers apoptosis when overexpressed in MCF-7 cells. We demonstrate that endogenous nCLU synthesis is a product of alternative splicing. Reverse transcriptase-PCR analyses revealed that exon II, containing the first AUG and encoding the endoplasmic reticulum-targeting peptide, was omitted. Exons I and III are spliced together placing a downstream AUG in exon III as the first available translation start site. This shorter mRNA produces the 49-kDa precursor nCLU protein. Ku70 binding activity was localized to the C-terminal coiled-coil domain of nCLU. Leucine residues 357, 358, and 361 of nCLU were necessary for Ku70-nCLU interaction. The N- and C-terminal coiled-coil domains of nCLU interacted with each other, suggesting that the protein could dimerize or fold. Mutation analyses indicate that the C-terminal NLS was functional in nCLU with the same contribution from N-terminal NLS. The C-terminal coiled-coil domain of nCLU was the minimal region required for Ku binding and apoptosis. MCF-7 cells show nuclear as well as cytoplasmic expression of GFP-nCLU in apoptotic cells. Cytosolic aggregation of GFP-nCLU was found in viable cells. These results indicate that an inactive precursor of nCLU exists in the cytoplasm of non irradiated MCF-7 cells, translocates into the nucleus following IR, and induces apoptosis. PMID- 12551934 TI - NFBD1, like 53BP1, is an early and redundant transducer mediating Chk2 phosphorylation in response to DNA damage. AB - Signaling pathways in response to DNA double strand breaks involve molecular cascades consisting of sensors, transducers, and effector proteins that activate cell cycle checkpoints and recruit repair machinery proteins. NFBD1 (a nuclear factor with BRCT domains protein 1) contains FHA (forkhead-associated), BRCT (breast cancer susceptibility gene 1 carboxyl terminus) domains, and internal repeats and is an early participant in nuclear foci in response to IR. To elucidate its role in the response pathways, small interfering RNA (siRNA) directed against NFDB1 in human cells demonstrated that its absence is associated with increased radio-sensitivity and delayed G(2)/M transition, but not G(1) to S. NFBD1 associates with nuclear foci within minutes following IR, a property similar to histone H2AX, 53BP1, and Chk2, which are all early participants in the DNA damage signaling cascade. Temporal studies show that H2AX is required for the foci positive for NFBD1, but NFBD1 is not needed for 53BP1- and H2AX-positive foci. NFBD1, together with 53BP1, plays a partially redundant role in regulating phosphorylation of the downstream effector protein, Chk2, since abrogation of both diminishes phosphorylated Chk2 in IR-induced foci. These results place NFBD1 parallel to 53BP1 in regulating Chk2 and downstream of H2AX in the recruitment of repair and signaling proteins to sites of DNA damage. PMID- 12551935 TI - Aggregate formation in Cu,Zn superoxide dismutase-related proteins. AB - Aggregation of Cu,Zn superoxide dismutase (SOD1) protein is a pathologic hallmark of familial amyotrophic lateral sclerosis linked to mutations in the SOD1 gene, although the structural motifs within mutant SOD1 that are responsible for its aggregation are unknown. Copper chaperone for SOD1 (CCS) and extracellular Cu,Zn superoxide dismutase (SOD3) have some sequence identity with SOD1, particularly in the regions of metal binding, but play no significant role in mutant SOD1 induced disease. We hypothesized that it would be possible to form CCS- or SOD3 positive aggregates by making these molecules resemble mutant SOD1 via the introduction of point mutations in codons homologous to a disease causing G85R SOD1 mutation. Using an in vitro assay system, we found that expression of wild type human CCS or a modified intracellular wild type SOD3 does not result in significant aggregate formation. In contrast, expression of G168R CCS or G146R SOD3 produced aggregates as evidenced by the presence of high molecular weight protein complexes on Western gels or inclusion bodies on immunofluorescence. CCS- and SOD3-positive inclusions appear to be ubiquitinated and localized to aggresomes. These results suggest that proteins sharing structural similarities to mutant SOD1 are also at risk for aggregate formation. PMID- 12551936 TI - Adipocyte low density lipoprotein receptor-related protein gene expression and function is regulated by peroxisome proliferator-activated receptor gamma. AB - The alpha(2)-macroglobulin receptor/low density lipoprotein receptor-related protein (LRP) is a large multifunctional receptor that interacts with a variety of molecules. It is implicated in biologically important processes such as lipoprotein metabolism, neurological function, tissue remodeling, protease complex clearance, and cell signal transduction. However, the regulation of LRP gene expression remains largely unknown. In this study, we have analyzed 2 kb of the 5'-flanking region of the LRP gene and identified a predicted peroxisome proliferator response element (PPRE) from -1185 to -1173. Peroxisome proliferator activated receptor gamma (PPARgamma) ligands such as fatty acids and rosiglitazone increased functional cell surface LRP by 1.5-2.0-fold in primary human adipocytes and in the SW872 human liposarcoma cell line as assessed by activated alpha(2)-macroglobulin binding and degradation. These agents were found to increase LRP transcription. Gel shift analysis of the putative PPRE demonstrated direct binding of PPARgamma/retinoid X receptor alpha heterodimers to the PPRE in the LRP gene. Furthermore, these heterodimers could no longer interact with a mutated PPRE probe. The isolated promoter was functional in SW872 cells, and its activity was increased by 1.5-fold with the addition of rosiglitazone. Furthermore, the isolated response element was similarly responsive to rosiglitazone when placed upstream of an ideal promoter. Mutagenesis of the predicted PPRE abolished the ability of this construct to respond to rosiglitazone. These data demonstrate that fatty acids and rosiglitazone directly stimulate transcription of the LRP gene through activation of PPARgamma and increase functional LRP expression. PMID- 12551937 TI - Regiospecific nitrosation of N-terminal-blocked tryptophan derivatives by N2O3 at physiological pH. AB - N(2)O(3) formed from nitric oxide in the presence of oxygen attacks thiols in proteins to yield S-nitrosothiols, which are believed to play a central role in NO signaling. In the present study we examined the N-nitrosation of N-terminal blocked (N-blocked) tryptophan derivatives in the presence of N(2)O(3) generating systems, such as preformed nitric oxide and nitric oxide donor compounds in the presence of oxygen at pH 7.4. Under these conditions N-nitrosation of N acetyltryptophan and lysine-tryptophan-lysine, respectively, was proven unequivocally by UV-visible spectroscopy as well as (15)N NMR spectrometry. Competition experiments performed with the known N(2)O(3) scavenger morpholine demonstrated that the selected tryptophan derivatives were nitrosated by N(2)O(3) with similar rate constants. It is further shown that the addition of ascorbate (vitamin C) induced the release of nitric oxide from N-acetyl-N-nitrosotryptophan as monitored polarographically with a NO electrode. Theoretical considerations strongly suggested that the reactivity of protein-bound tryptophan would be high enough to compete effectively with protein-bound cysteine for N(2)O(3). Our data demonstrate conclusively that N(2)O(3) nitrosates the secondary amine function (N(indole)) at the indole ring of N-blocked tryptophan with high reactivity at physiological pH values. PMID- 12551938 TI - ATP binding to Rho transcription termination factor. Mutant F355W ATP-induced fluorescence quenching reveals dynamic ATP binding. AB - Rho transcription termination factor mutant, F355W, showed tryptophan fluorescence intensity approximately twice that of wild-type Rho at equivalent protein concentrations and underwent a decrease in relative fluorescence intensity at 350 nm when 100 microm ATP was added in the presence or absence of RNA. Titration of this fluorescence quenching with varying concentrations of ATP (0-600 microm), where Rho is shown to exist as a hexamer (400 nm Rho), revealed tight and loose ATP-binding sites. Bicyclomycin, a specific inhibitor of Rho, increased the tight ATP binding and was used to calibrate ATP-induced fluorescence quenching by using [gamma-(32)P]ATP filter binding. For the Rho mutant F355W, three tight (K(d)(1) = 3 +/- 0.3 microm) and three loose (K(d)(2) = 58 +/- 3 microm) ATP-binding sites per hexamer were seen on Scatchard analysis in the absence of bicyclomycin and poly(C). In the presence of bicyclomycin, the K(d)(1) changed from 3.0 to 1.4 microm, but K(d)(2) underwent a lesser change. The non-hydrolyzable ATP analogue, gamma-S-ATP, gave a similar profile with three tight (K(d)(1) = 0.2 microm) and three loose (K(d)(2) = 70 microm) ATP-binding sites per hexamer. Adding poly(C) to F355W did not alter the K(d)(1) or K(d)(2) for ATP or for gamma-S-ATP. ADP-induced quenching produced 5.5 loose (K(d) = 92 microm) binding sites in the absence of poly(C), and the binding became weaker (K(d) = 175 microm) in the presence of poly(C). The data suggest that in the presence of ADP Rho has six equivalent nucleotide-binding sites. When ATP was added these sites converted to three tight and three loose binding loci. We propose an alternating ATP site mechanism where ATP binding creates heterogeneity in the ATP binding in adjacent subunits, and we suggest that ATP binding to a neighboring loose site stimulates hydrolysis at a neighboring tight binding site such that all six subunits can be potential "active" sites for ATP hydrolysis. The dynamic nature of the ATP binding to Rho is discussed in the terms of the mechanism of RNA tracking driven by ATP hydrolysis. PMID- 12551939 TI - Activation of orphan nuclear constitutive androstane receptor requires subnuclear targeting by peroxisome proliferator-activated receptor gamma coactivator-1 alpha. A possible link between xenobiotic response and nutritional state. AB - In contrast to the classical nuclear receptors, the constitutive androstane receptor (CAR) is transcriptionally active in the absence of ligand. In the course of searching for the mediator of CAR activation, we found that ligand independent activation of CAR was achieved in cooperation with the peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1 alpha). PGC-1 beta, a PGC-1 alpha homologue, also activated CAR to less of an extent than PGC-1 alpha. Coexpression of the ligand-binding domain of a heterodimerization partner, retinoid X receptor alpha, enhanced the PGC-1 alpha-mediated activation of CAR, although it had a weak effect on the basal activity of CAR in the absence of PGC 1 alpha. Both the N-terminal region, with the LXXLL motif, and the C-terminal region, with a serine/arginine-rich domain (RS domain), in PGC-1 alpha were required for full activation of CAR. Pull-down experiments using recombinant proteins revealed that CAR directly interacted with both the LXXLL motif and the RS domain. Furthermore, we demonstrated that the RS domain of PGC-1 alpha was required for CAR localization at nuclear speckles. These results indicate that PGC-1 alpha mediates the ligand-independent activation of CAR by means of subnuclear targeting through the RS domain of PGC-1 alpha. PMID- 12551940 TI - Hepatocyte-derived ApoE is more effective than non-hepatocyte-derived ApoE in remnant lipoprotein clearance. AB - The importance of hepatocyte-derived apolipoprotein (apo) E in the clearance of remnant lipoproteins in the liver is controversial. To address this controversy, we compared remnant clearance in two mouse models in which apoE is primarily derived either from hepatocytes or from an extrahepatic source. Hypomorphic apoE mice universally express reduced levels of apoE in all tissues, with the liver remaining the primary source of apoE. This mouse model of hepatocyte-derived apoE was compared with Apoe(-/-) mice transplanted with mouse bone marrow as a model of primarily non-hepatocyte-derived apoE. Immunohistochemical analysis of liver sections revealed that only the hepatocyte-derived apoE model had detectable levels of apoE on hepatic sinusoidal surfaces. The non-hepatocyte-derived apoE model with plasma apoE levels similar to those in the hepatocyte-derived model had 2-fold more total plasma cholesterol, 4-fold more total plasma triglycerides, and 8-fold higher levels of apoB48, similar to Apoe(-/-) mice. Both the hepatocyte-derived and the non-hepatocyte-derived apoE models had delayed clearance of an infused bolus of (125)I-labeled remnants compared with wild-type mice. However, after 3 h, plasma remnants reached wild-type levels only in the hepatocyte-derived apoE model, which had accumulated 70 +/- 5% of wild-type levels of remnants in the liver while the non-hepatocyte-derived apoE model had accumulated only 38 +/- 4%. These results demonstrate the existence of a role for both hepatically derived and localized apoE in remnant clearance. This role likely represents the enrichment of remnants sequestered on hepatocyte, with hepatocyte-derived apoE, facilitating their receptor-mediated internalization. PMID- 12551941 TI - Location of the actual signal in the negatively charged leader sequence involved in the import into the mitochondrial matrix space. AB - Proteins destined for the mitochondrial matrix space have leader sequences that are typically present at the most N-terminal end of the nuclear-encoded precursor protein. The leaders are rich in positive charges and usually deficient of negative charges. This observation led to the acid-chain hypothesis to explain how the leader sequences interact with negatively charged receptor proteins. Here we show using both chimeric leaders and one from isopropyl malate synthase that possesses a negative charge that the leader need not be at the very N terminus of the precursor. Experiments were performed with modified non-functioning leader sequences fused to either the native or a non-functioning leader of aldehyde dehydrogenase so that an internal leader sequence could exist. The internal leader is sufficient for the import of the modified precursor protein. It appears that this leader still needs to form an amphipathic helix just like the normal N terminal leaders do. This internal leader could function even if the most N terminal portion contained negative charges in the first 7-11 residues. If the first 11 residues were deleted from isopropyl malate synthase, the resulting protein was imported more successfully than the native protein. It appears that precursors that carry negatively charged leaders use an internal signal sequence to compensate for the non-functional segment at the most N-terminal portion of the protein. PMID- 12551942 TI - Self-association of the spindle pole body-related intermediate filament protein Fin1p and its phosphorylation-dependent interaction with 14-3-3 proteins in yeast. AB - The Fin1 protein of the yeast Saccharomyces cerevisiae forms filaments between the spindle pole bodies of dividing cells. In the two-hybrid system it binds to 14-3-3 proteins, which are highly conserved proteins involved in many cellular processes and which are capable of binding to more than 120 different proteins. Here, we describe the interaction of the Fin1 protein with the 14-3-3 proteins Bmh1p and Bmh2p in more detail. Purified Fin1p interacts with recombinant yeast 14-3-3 proteins. This interaction is strongly reduced after dephosphorylation of Fin1p. Surface plasmon resonance analysis showed that Fin1p has a higher affinity for Bmh2p than for Bmh1p (K(D) 289 versus 585 nm). Sequences in both the central and C-terminal part of Fin1p are required for the interaction with Bmh2p in the two-hybrid system. In yeast strains lacking 14-3-3 proteins Fin1 filament formation was observed, indicating that the 14-3-3 proteins are not required for this process. Fin1 also interacts with itself in the two-hybrid system. For this interaction sequences at the C terminus, containing one of two putative coiled coil regions, are sufficient. Fin1p-Fin1p interactions were demonstrated in vivo by fluorescent resonance energy transfer between cyan fluorescent protein-labeled Fin1p and yellow fluorescent protein-labeled Fin1p. PMID- 12551943 TI - The distribution of lipoprotein lipase in rat adipose tissue. Changes with nutritional state engage the extracellular enzyme. AB - Lipoprotein lipase (LPL) acts at the vascular endothelium. Earlier studies have shown that down-regulation of adipose tissue LPL during fasting is post translational and involves a shift from active to inactive forms of the lipase. Studies in cell systems had indicated that during fasting LPL might be retained in the endoplasmic reticulum. We have now explored the relation between active/inactive and intra/extracellular forms of the lipase. Within adipocytes, neither LPL mass nor the distribution of LPL between active and inactive forms changed on fasting. Extracellular LPL mass also did not change significantly, but shifted from predominantly active to predominantly inactive. To explore if changes in secretion were compensated by changes in turnover, synthesis of new protein was blocked by cycloheximide. The rates at which intra- and extracellular LPL mass and activity decreased did not change on fasting. To further explore how LPL is distributed in the tissue, heparin (which detaches the enzyme from the endothelial surface) was injected. Tissue LPL activity decreased by about 10% in 2 min and by 50% in 1 h. Heparin released mainly the active form of the lipase. There was no change of LPL activity or mass within adipocytes. The fraction of extracellular LPL that heparin released and the time course were the same in fed and fasted rats, indicating that active, extracellular LPL was distributed in a similar way in the two nutritional states. This study suggests that the nutritional regulation of LPL in adipose tissue determines the activity state of extracellular LPL. PMID- 12551944 TI - C termini of the Escherichia coli mechanosensitive ion channel (MscS) move apart upon the channel opening. AB - Heptameric YggB is a mechanosensitive ion channel (MscS) from the inner membrane of Escherichia coli. We demonstrate, using the patch clamp technique, that cross linking of the YggB C termini led to irreversible inhibition of the channel activities. Application of Ni(2+) to the YggB-His(6) channels with the hexahistidine tags added to the ends of their C termini also resulted in a marked but reversible decrease of activities. Western blot revealed that YggB-His(6) oligomers are more stable in the presence of Ni(2+), providing evidence that Ni(2+) is coordinated between C termini from different subunits of the channel. Intersubunit coordination of Ni(2+) affecting channel activities occurred in the channel closed conformation and not in the open state. This may suggest that the C termini move apart upon channel opening and are involved in the channel activation. We propose that the as yet undefined C-terminal region may form a cytoplasmic gate of the channel. The results are discussed and interpreted based on the recently released quaternary structure of the channel. PMID- 12551945 TI - ABCA1 is essential for efficient basolateral cholesterol efflux during the absorption of dietary cholesterol in chickens. AB - The ATP-binding cassette transporter A1 (ABCA1) participates in the efflux of cholesterol from cells. It remains unclear whether ABCA1 functions to efflux cholesterol across the basolateral or apical membrane of the intestine. We used a chicken model of ABCA1 dysfunction, the Wisconsin hypoalpha mutant (WHAM) chicken, to address this issue. After an oral gavage of radioactive cholesterol, the percentage appearing in the bloodstream was reduced by 79% in the WHAM chicken along with a 97% reduction in the amount of tracer in high density lipoprotein. In contrast, the percentage of radioactive cholesterol absorbed from the lumen into the intestine was not affected by the ABCA1 mutation. Liver X receptor (LXR) agonists have been inferred to decrease cholesterol absorption through activation of ABCA1 expression. However, the LXR agonist T0901317 decreased cholesterol absorption equally in both wild type and WHAM chickens, indicating that the effect of LXR activation on cholesterol absorption is independent of ABCA1. The ABCA1 mutation resulted in accumulation of radioactive cholesterol ester in the intestine and the liver of the WHAM chicken (5.0- and 4.4-fold, respectively), whereas biliary lipid concentrations were unaltered by the WHAM mutation. In summary, ABCA1 regulates the efflux of cholesterol from the basolateral but not apical membrane in the intestine and the liver. PMID- 12551946 TI - Targeted disruption of intracellular type I platelet activating factor acetylhydrolase catalytic subunits causes severe impairment in spermatogenesis. AB - Intracellular type I platelet activating factor-acetylhydrolase is a phospholipase that consists of a dimer of two homologous catalytic subunits alpha1 and alpha2 as well as LIS1, a product of the causative gene for type I lissencephaly. LIS1 plays an important role in neuronal migration during brain development, but the in vivo function of the catalytic subunits remains unclear. In this study, we generated alpha1- and a2-deficient mice by targeted disruption. alpha1(-/-) mice are indistinguishable from wild-type mice, whereas alpha2(-/-) male mice show a significant reduction in testis size. Double-mutant male mice are sterile because of severe impairment of spermatogenesis. Histological examination revealed marked degeneration at the spermatocyte stage and an increase of apoptotic cells in the seminiferous tubules. The catalytic subunits are expressed at high levels in testis as well as brain in mice. In wild-type mice, alpha2 is expressed in all seminiferous tubule cell types, whereas alpha1 is expressed only in the spermatogonia. This expression pattern parallels the finding that deletion of both subunits induces a marked loss of germ cells at an early spermatogenic stage. We also found that the LIS1 protein levels, but not the mRNA levels, were significantly reduced in alpha2(-/-) and double-mutant mice, suggesting that the catalytic subunits, especially alpha2, are a determinant of LIS1 expression level. PMID- 12551947 TI - Both Max and TFE3 cooperate with Smad proteins to bind the plasminogen activator inhibitor-1 promoter, but they have opposite effects on transcriptional activity. AB - Transforming growth factor (TGF)-beta regulates gene expression in large part through combinatorial interactions between members of the Smad family and other transcription factors. The basic helix-loop-helix leucine zipper (bHLHZIP) protein TFE3 and Smad3 synergistically activate transcription of the plasminogen activator inhibitor-1 (PAI-1) as well as other genes. We investigated interactions among different bHLHZIP and Smad family proteins. TFE3, TFEB, and Max associated with Smad3 and Smad4 in the absence of DNA and at the PE2.1 element of the PAI-1 promoter. These interactions were mediated by the leucine zipper and MH1 regions of the respective proteins. No interactions were observed with the E47 bHLH family protein. Chimeric proteins, in which leucine zippers from bHLHZIP or bZIP proteins were fused to heterologous bHLH domains, associated with Smad proteins both in the absence of DNA and at the PE2.1 element. The kinetics of bHLHZIP and Smad protein binding at the PE2.1 element were examined using surface plasmon resonance analysis. TFE3 exhibited cooperative DNA binding with Smad proteins, whereas no cooperativity was observed between E47 and Smads. Max inhibited transcription activation by Smad3 and TGF-beta at the PAI-1 promoter, whereas TFE3 and TFEB stimulated transcription activation. These results suggest that Smad family proteins can interact with several bHLHZIP proteins, resulting in different transcriptional outcomes. PMID- 12551948 TI - 14-3-3 connects glycogen synthase kinase-3 beta to tau within a brain microtubule associated tau phosphorylation complex. AB - In a recent study, we reported that in bovine brain extract, glycogen synthase kinase-3beta and tau are parts of an approximately 400-500 kDa microtubule associated tau phosphorylation complex (Sun, W., Qureshi, H. Y., Cafferty, P. W., Sobue, K., Agarwal-Mawal, A., Neufield, K. D., and Paudel, H. K. (2002) J. Biol. Chem. 277, 11933-11940). In this study, we find that when purified brain microtubules are subjected to Superose 12 gel filtration column chromatography, the dimeric scaffold protein 14-3-3 zeta co-elutes with the tau phosphorylation complex components tau and GSK3 beta. From gel filtration fractions containing the tau phosphorylation complex, 14-3-3 zeta, GSK3 beta, and tau co immunoprecipitate with each other. From extracts of bovine brain, COS-7 cells, and HEK-293 cells transfected with GSK3 beta, 14-3-3 zeta co-precipitates with GSK3 beta, indicating that GSK3 beta binds to 14-3-3 zeta. From HEK-293 cells transfected with tau, GSK3 beta, and 14-3-3 zeta in different combinations, tau co-immunoprecipitates with GSK3 beta only in the presence of 14-3-3 zeta. In vitro, approximately 10-fold more tau binds to GSK3 beta in the presence of than in the absence of 14-3-3 zeta. In transfected HEK-293 cells, 14-3-3 zeta stimulates GSK3 beta-catalyzed tau phosphorylation in a dose-dependent manner. These data indicate that in brain, the 14-3-3 zeta dimer simultaneously binds and bridges tau and GSK3 beta and stimulates GSK3 beta-catalyzed tau phosphorylation. PMID- 12551949 TI - Lymphoid enhancer factor-1 and beta-catenin inhibit Runx2-dependent transcriptional activation of the osteocalcin promoter. AB - Functional control of the transcription factor Runx2 is crucial for normal bone formation. Runx2 is detectable throughout osteoblast development and maturation and temporally regulates several bone-specific genes. In this study, we identified a novel post-translational mechanism regulating Runx2-dependent activation of the osteocalcin promoter. A functional binding site for the high mobility group protein lymphoid enhancer-binding factor 1 (LEF1) was found adjacent to the proximal Runx2-binding site in the osteocalcin promoter. In transcription assays, LEF1 repressed Runx2-induced activation of the mouse osteocalcin 2 promoter in several osteoblast lineage cell lines. Mutations in the LEF1-binding site increased the basal activity of the osteocalcin promoter; however, the LEF1 recognition site in the osteocalcin promoter was surprisingly not required for LEF1 repression. A novel interaction between the DNA-binding domains of Runx2 and LEF1 was identified and found crucial for LEF1-mediated repression of Runx2. LEF1 is a nuclear effector of the Wnt/LRP5/beta-catenin signaling pathway, which is also essential for osteoblast proliferation and normal skeletal development. A constitutively active beta-catenin enhanced LEF1 dependent repression of Runx2. These data identify a novel mechanism of regulating Runx2 activity in osteoblasts and link Runx2 transcriptional activity to beta-catenin signaling. PMID- 12551950 TI - Candida albicans phospholipomannan promotes survival of phagocytosed yeasts through modulation of bad phosphorylation and macrophage apoptosis. AB - The surface of the pathogenic yeast Candida albicans is coated with phospholipomannan (PLM), a phylogenetically unique glycolipid composed of beta 1,2-oligomannosides and phytoceramide. This study compared the specific contribution of PLM to the modulation of signaling pathways linked to the survival of C. albicans in macrophages in contrast to Saccharomyces cerevisiae. C. albicans endocytosis by J774 and disregulation of the ERK1/2 signal transduction pathway was associated downstream with a reduction in Bad Ser-112 phosphorylation and disappearance of free Bcl-2. This suggested an apoptotic effect, which was confirmed by staining of phosphatidylserine in the macrophage outer membrane. The addition of PLM to macrophages incubated with S. cerevisiae mimicked each of the disregulation steps observed with C. albicans and promoted the survival of S. cerevisiae. Externalization of membranous phosphatidylserine, loss of mitochondrial integrity, and DNA fragmentation induced by PLM showed that this molecule promoted yeast survival by inducing host cell death. These findings suggest strongly that PLM is a virulence attribute of C. albicans and that elucidation of the relationship between structure and apoptotic activity is an innovative field of research. PMID- 12551951 TI - Drosophila p120catenin plays a supporting role in cell adhesion but is not an essential adherens junction component. AB - Cadherin-catenin complexes, localized to adherens junctions, are essential for cell-cell adhesion. One means of regulating adhesion is through the juxtamembrane domain of the cadherin cytoplasmic tail. This region is the binding site for p120, leading to the hypothesis that p120 is a key regulator of cell adhesion. p120 has also been suggested to regulate the GTPase Rho and to regulate transcription via its binding partner Kaiso. To test these hypothesized functions, we turned to Drosophila, which has only a single p120 family member. It localizes to adherens junctions and binds the juxtamembrane region of DE cadherin (DE-cad). We generated null alleles of p120 and found that mutants are viable and fertile and have no substantial changes in junction structure or function. However, p120 mutations strongly enhance mutations in the genes encoding DE-cadherin or Armadillo, the beta-catenin homologue. Finally, we examined the localization of p120 during embryogenesis. p120 localizes to adherens junctions, but its localization there is less universal than that of core adherens junction proteins. Together, these data suggest that p120 is an important positive modulator of adhesion but that it is not an essential core component of adherens junctions. PMID- 12551952 TI - Rae1 is an essential mitotic checkpoint regulator that cooperates with Bub3 to prevent chromosome missegregation. AB - The WD-repeat proteins Rae1 and Bub3 show extensive sequence homology, indicative of functional similarity. However, previous studies have suggested that Rae1 is involved in the mRNA export pathway and Bub3 in the mitotic checkpoint. To determine the in vivo roles of Rae1 and Bub3 in mammals, we generated knockout mice that have these genes deleted individually or in combination. Here we show that haplo-insufficiency of either Rae1 or Bub3 results in a similar phenotype involving mitotic checkpoint defects and chromosome missegregation. We also show that overexpression of Rae1 can correct for Rae1 haplo-insufficiency and, surprisingly, Bub3 haplo-insufficiency. Rae1-null and Bub3-null mice are embryonic lethal, although cells from these mice did not have a detectable defect in nuclear export of mRNA. Unlike null mice, compound haplo-insufficient Rae1/Bub3 mice are viable. However, cells from these mice exhibit much greater rates of premature sister chromatid separation and chromosome missegregation than single haplo-insufficient cells. Finally, we show that mice with mitotic checkpoint defects are more susceptible to dimethylbenzanthrene-induced tumorigenesis than wild-type mice. Thus, our data demonstrate a novel function for Rae1 and characterize Rae1 and Bub3 as related proteins with essential, overlapping, and cooperating roles in the mitotic checkpoint. PMID- 12551953 TI - Anthrax toxin triggers endocytosis of its receptor via a lipid raft-mediated clathrin-dependent process. AB - The protective antigen (PA) of the anthrax toxin binds to a cell surface receptor and thereby allows lethal factor (LF) to be taken up and exert its toxic effect in the cytoplasm. Here, we report that clustering of the anthrax toxin receptor (ATR) with heptameric PA or with an antibody sandwich causes its association to specialized cholesterol and glycosphingolipid-rich microdomains of the plasma membrane (lipid rafts). We find that although endocytosis of ATR is slow, clustering it into rafts either via PA heptamerization or using an antibody sandwich is necessary and sufficient to trigger efficient internalization and allow delivery of LF to the cytoplasm. Importantly, altering raft integrity using drugs prevented LF delivery and cleavage of cytosolic MAPK kinases, suggesting that lipid rafts could be therapeutic targets for drugs against anthrax. Moreover, we show that internalization of PA is dynamin and Eps15 dependent, indicating that the clathrin-dependent pathway is the major route of anthrax toxin entry into the cell. The present work illustrates that although the physiological role of the ATR is unknown, its trafficking properties, i.e., slow endocytosis as a monomer and rapid clathrin-mediated uptake on clustering, make it an ideal anthrax toxin receptor. PMID- 12551954 TI - Dynactin is required for bidirectional organelle transport. AB - Kinesin II is a heterotrimeric plus end-directed microtubule motor responsible for the anterograde movement of organelles in various cell types. Despite substantial literature concerning the types of organelles that kinesin II transports, the question of how this motor associates with cargo organelles remains unanswered. To address this question, we have used Xenopus laevis melanophores as a model system. Through analysis of kinesin II-mediated melanosome motility, we have determined that the dynactin complex, known as an anchor for cytoplasmic dynein, also links kinesin II to organelles. Biochemical data demonstrates that the putative cargo-binding subunit of Xenopus kinesin II, Xenopus kinesin II-associated protein (XKAP), binds directly to the p150Glued subunit of dynactin. This interaction occurs through aa 530-793 of XKAP and aa 600-811 of p150Glued. These results reveal that dynactin is required for transport activity of microtubule motors of opposite polarity, cytoplasmic dynein and kinesin II, and may provide a new mechanism to coordinate their activities. PMID- 12551955 TI - Rac and Cdc42 play distinct roles in regulating PI(3,4,5)P3 and polarity during neutrophil chemotaxis. AB - Neutrophils exposed to chemoattractants polarize and accumulate polymerized actin at the leading edge. In neutrophil-like HL-60 cells, this asymmetry depends on a positive feedback loop in which accumulation of a membrane lipid, phosphatidylinositol (PI) 3,4,5-trisphosphate (PI[3,4,5]P3), leads to activation of Rac and/or Cdc42, and vice versa. We now report that Rac and Cdc42 play distinct roles in regulating this asymmetry. In the absence of chemoattractant, expression of constitutively active Rac stimulates accumulation at the plasma membrane of actin polymers and of GFP-tagged fluorescent probes for PI(3,4,5)P3 (the PH domain of Akt) and activated Rac (the p21-binding domain of p21-activated kinase). Dominant negative Rac inhibits chemoattractant-stimulated accumulation of actin polymers and membrane translocation of both fluorescent probes and attainment of morphologic polarity. Expression of constitutively active Cdc42 or of two different protein inhibitors of Cdc42 fails to mimic effects of the Rac mutants on actin or PI(3,4,5)P3. Instead, Cdc42 inhibitors prevent cells from maintaining a persistent leading edge and frequently induce formation of multiple, short lived leading edges containing actin polymers, PI(3,4,5)P3, and activated Rac. We conclude that Rac plays a dominant role in the PI(3,4,5)P3 dependent positive feedback loop required for forming a leading edge, whereas location and stability of the leading edge are regulated by Cdc42. PMID- 12551956 TI - Binding site for p120/delta-catenin is not required for Drosophila E-cadherin function in vivo. AB - Homophilic cell adhesion mediated by classical cadherins is important for many developmental processes. Proteins that interact with the cytoplasmic domain of cadherin, in particular the catenins, are thought to regulate the strength and possibly the dynamics of adhesion. beta-catenin links cadherin to the actin cytoskeleton via alpha-catenin. The role of p120/delta-catenin proteins in regulating cadherin function is less clear. Both beta-catenin and p120/delta catenin are conserved in Drosophila. Here, we address the importance of cadherin catenin interactions in vivo, using mutant variants of Drosophila epithelial cadherin (DE-cadherin) that are selectively defective in p120ctn (DE-cadherin AAA) or beta-catenin-armadillo (DE-cadherin-Delta beta) interactions. We have analyzed the ability of these proteins to substitute for endogenous DE-cadherin activity in multiple cadherin-dependent processes during Drosophila development and oogenesis; epithelial integrity, follicle cell sorting, oocyte positioning, as well as the dynamic adhesion required for border cell migration. As expected, DE-cadherin-Delta beta did not substitute for DE-cadherin in these processes, although it retained some residual activity. Surprisingly, DE-cadherin-AAA was able to substitute for the wild-type protein in all contexts with no detectable perturbations. Thus, interaction with p120/delta-catenin does not appear to be required for DE-cadherin function in vivo. PMID- 12551957 TI - A biomimetic motility assay provides insight into the mechanism of actin-based motility. AB - Abiomimetic motility assay is used to analyze the mechanism of force production by site-directed polymerization of actin. Polystyrene microspheres, functionalized in a controlled fashion by the N-WASP protein, the ubiquitous activator of Arp2/3 complex, undergo actin-based propulsion in a medium that consists of five pure proteins. We have analyzed the dependence of velocity on N WASP surface density, on the concentration of capping protein, and on external force. Movement was not slowed down by increasing the diameter of the beads (0.2 to 3 microm) nor by increasing the viscosity of the medium by 10(5)-fold. This important result shows that forces due to actin polymerization are balanced by internal forces due to transient attachment of filament ends at the surface. These forces are greater than the viscous drag. Using Alexa488-labeled Arp2/3, we show that Arp2/3 is incorporated in the actin tail like G-actin by barbed end branching of filaments at the bead surface, not by side branching, and that filaments are more densely branched upon increasing gelsolin concentration. These data support models in which the rates of filament branching and capping control velocity, and autocatalytic branching of filament ends, rather than filament nucleation, occurs at the particle surface. PMID- 12551958 TI - Molecular biology of early olfactory memory. PMID- 12551959 TI - Mitral cell beta1 and 5-HT2A receptor colocalization and cAMP coregulation: a new model of norepinephrine-induced learning in the olfactory bulb. AB - In the present study we assess a new model for classical conditioning of odor preference learning in rat pups. In preference learning beta(1)-adrenoceptors activated by the locus coeruleus mediate the unconditioned stimulus, whereas olfactory nerve input mediates the conditioned stimulus, odor. Serotonin (5-HT) depletion prevents odor learning, with 5-HT(2A/2C) agonists correcting the deficit. Our new model proposes that the interaction of noradrenergic and serotonergic input with odor occurs in the mitral cells of the olfactory bulb through activation of cyclic adenosine monophosphate (cAMP). Here, using selective antibodies and immunofluorescence examined with confocal microscopy, we demonstrate that beta(1)-adrenoceptors and 5-HT(2A) receptors colocalize primarily on mitral cells. Using a cAMP assay and cAMP immunocytochemistry, we find that beta-adrenoceptor activation by isoproterenol, at learning-effective and higher doses, significantly increases bulbar cAMP, as does stroking. As predicted by our model, the cAMP increases are localized to mitral cells. 5-HT depletion of the olfactory bulb does not affect basal levels of cAMP but prevents isoproterenol-induced cAMP elevation. These results support the model. We suggest the mitral-cell cAMP cascade converges with a Ca(2+) pathway activated by odor to recruit CREB phosphorylation and memory-associated changes in the olfactory bulb. The dose-related increase in cAMP with isoproterenol implies a critical cAMP window because the highest dose of isoproterenol does not produce learning. PMID- 12551960 TI - Conflicting processes in the extinction of conditioned taste aversion: behavioral and molecular aspects of latency, apparent stagnation, and spontaneous recovery. AB - The study of experimental extinction and of the spontaneous recovery of the extinguished memory could cast light on neurobiological mechanisms by which internal representations compete to control behavior. In this work, we use a combination of behavioral and molecular methods to dissect subprocesses of experimental extinction of conditioned taste aversion (CTA). Extinction of CTA becomes apparent only 90 min after the extinction trial. This latency is insensitive to muscarinic and beta-adrenergic modulation and to protein synthesis inhibition in the insular cortex (IC). Immediately afterwards, however, the extinguishing trace becomes sensitive to beta-adrenergic blockade and protein synthesis inhibition. The subsequent kinetics and magnitude of extinction depend on whether a spaced or massed extinction protocol is used. A massed protocol is highly effective in the short run, but results in apparent stagnation of extinction in the long-run, which conceals fast spontaneous recovery of the preextinguished trace. This recovery can be truncated by a beta-adrenergic agonist or a cAMP analog in the insular cortex, suggesting that spontaneous overtaking of the behavioral control by the original association is regulated at least in part by beta-adrenergic input, probably operating via the cAMP cascade, long after the offset of the conditioned stimulus. Hence, the performance of the subject in experimental extinction is the sum total of multiple, sometimes conflicting, time-dependent processes. PMID- 12551961 TI - A role for ERK MAP kinase in physiologic temporal integration in hippocampal area CA1. AB - Recent studies demonstrate a requirement for the Extracellular signal Regulated Kinase (ERK) mitogen-activated protein kinase (MAPK) cascade in both the induction of long-lasting forms of hippocampal synaptic plasticity and in hippocampus-dependent associative and spatial learning. In the present studies, we investigated mechanisms by which ERK might contribute to synaptic plasticity at Schaffer collateral synapses in hippocampal slices. We found that long-term potentiation (LTP) induced with a pair of 100-Hz tetani does not require ERK activation in mice whereas it does in rats. However, in mice, inhibition of ERK activation blocked LTP induced by two LTP induction paradigms that mimicked the endogenous theta rhythm. In an additional series of studies, we found that mice specifically deficient in the ERK1 isoform of MAPK showed no impairments in tests of hippocampal physiology. To investigate ERK-dependent mechanisms operating during LTP-inducing stimulation paradigms, we monitored spike production in the cell body layer of the hippocampus during the period of theta-like LTP-inducing stimulation. Theta-burst stimulation (TBS) produced a significant amount of postsynaptic spiking, and the likelihood of spike production increased progressively over the course of the three trains of TBS independent of any apparent increase in Excitatory Post-Synaptic Potential (EPSP) magnitude. Inhibition of ERK activation dampened this TBS-associated increase in spiking. These data indicate that, for specific patterns of stimulation, ERK may function in the regulation of neuronal excitability in hippocampal area CA1. Overall, our data indicate that the progressive increase in spiking observed during TBS represents a form of physiologic temporal integration that is dependent on ERK MAPK activity. PMID- 12551962 TI - Using an aplysia two-hybrid system to examine the interactions between transcription factors involved in long-term facilitation in the nervous system of aplysia. AB - Interactions between ApCREB1a, ApCREB2, and ApC/EBP have been studied using conventional methods such as the yeast two-hybrid system. However, it is unclear whether these memory-related transcription factors actually interact in the native environment in neurons. To clarify this question, we have developed an Aplysia two-hybrid system and found, consistent with previous studies that ApCREB2 interacts with ApCREB1a and ApC/EBP, and that ApC/EBP forms homodimers. We have also found that ApCREB1a and ApC/EBP do not interact. Therefore, our study shows that formerly described interactions between the proteins actually occur in the Aplysia neurons and that interactions between these transcription factors are specific. PMID- 12551963 TI - Long-term memory survives nerve injury and the subsequent regeneration process. AB - A three-neuron network (a central pattern generator [CPG]) is both sufficient and necessary to generate aerial respiratory behavior in the pond snail, Lymnaea stagnalis. Aerial respiratory behavior is abolished following a specific nerve crush that results in axotomy to one of the three CPG neurons, RPeD1. Functional regeneration of the crushed neurite occurs within 10 days, allowing aerial respiratory behavior to be restored. Functional regeneration does not occur if the connective is cut rather than crushed. In unaxotomized snails, aerial respiratory behavior can be operantly conditioned, and following memory consolidation, long-term memory (LTM) persists for at least 2 weeks. We used the Lymnaea model system to determine (1) If in naive animals axotomy and the subsequent regeneration result in a nervous system that is competent to mediate associative learning and LTM, and (2) if LTM survives RPeD1 axotomy and the subsequent regenerative process. We show here that (1) A regenerated nervous system is competent to mediate associative memory and LTM, and (2) LTM survives axotomy and the subsequent regenerative process. PMID- 12551964 TI - Visuomotor adaptation in normal aging. AB - Visuomotor adaptation to a gradual or sudden screen cursor rotation was investigated in healthy young and elderly subjects. Both age groups were equally divided into two subgroups; one subgroup was exposed to 11.25 degrees step increments of visual feedback rotation, every 45 trials (up to a total of 90 degrees), whereas a second subgroup was subjected to 90 degrees rotation from the onset of exposure. Participants performed discrete, horizontal hand movements to virtual targets in four randomized directions. Targets appeared on a computer screen in front of them, and a board prevented vision of the hand at all times. Differential effects of aging on visuomotor adaptation were found, depending on the time course of the visual distortion. In both age groups, early exposure to the sudden visual feedback distortion resulted in typical spiral-like trajectories, which became straighter by late exposure. However, the final adaptation level was reduced in the aged group, although the aftereffects were similar. When subjects were exposed to the gradual distortion, no statistically significant differences in measures of adaptation with advancing age were found. In this case, both age groups appeared to adapt equally. However, after removal of the distortion, elderly subjects showed reduced aftereffects as compared with the young group. These findings suggest differential effects of aging on adaptation to gradual versus sudden visual feedback distortions, and may help to explain the conflicting results obtained in previous visuomotor adaptation studies. PMID- 12551965 TI - Size and reversal learning in the beagle dog as a measure of executive function and inhibitory control in aging. AB - Several studies converge on the idea that executive processes age earlier than other cognitive processes. As part of a larger effort to investigate age-related changes in executive processes in the dog, inhibitory control was measured in young, middle-aged, old, and senior dogs using size discrimination learning and reversal procedures. Compared to young and middle-aged dogs, old and senior dogs were impaired on both the initial learning of the size task and the reversal of original reward contingencies. Impaired performance in the two aged groups was characterized as a delay in learning the correct stimulus-reward contingencies and, among the senior dogs in particular, an increase in perseverative responding. These separate patterns of reversal impairments in the old and senior dogs may reflect different rates of aging in subregions of the frontal cortex. PMID- 12551966 TI - Audiospatial and visuospatial working memory in 6-13 year old school children. AB - The neural processes subserving working memory, and brain structures underlying this system, continue to develop during childhood. We investigated the effects of age and gender on audiospatial and visuospatial working memory in a nonclinical sample of school-aged children using n-back tasks. The results showed that auditory and visual working memory performance improves with age, suggesting functional maturation of underlying cognitive processes and brain areas. The gender differences found in the performance of working memory tasks suggest a larger degree of immaturity in boys than girls at the age period of 6-10 yr. The differences observed between the mastering of auditory and visual working memory tasks may indicate that visual working memory reaches functional maturity earlier than the corresponding auditory system. PMID- 12551967 TI - Modification of the 5' terminus of Sindbis virus genomic RNA allows nsP4 RNA polymerases with nonaromatic amino acids at the N terminus to function in RNA replication. AB - We have previously shown that Sindbis virus RNA polymerase requires an N-terminal aromatic amino acid or histidine for wild-type or pseudo-wild-type function; mutant viruses with a nonaromatic amino acid at the N terminus of the polymerase, but which are otherwise wild type, are unable to produce progeny viruses and will not form a plaque at any temperature tested. We now show that such mutant polymerases can function to produce progeny virus sufficient to form plaques at both 30 and 40 degrees C upon addition of AU, AUA, or AUU to the 5' terminus of the genomic RNA or upon substitution of A for U as the third nucleotide of the genome. These results are consistent with the hypothesis that (i) 3'-UA-5' is required at the 3' terminus of the minus-strand RNA for initiation of plus-strand genomic RNA synthesis; (ii) in the wild-type virus this sequence is present in a secondary structure that can be opened by the wild-type polymerase but not by the mutant polymerase; (iii) the addition of AU, AUA, or AUU to the 5' end of the genomic RNA provides unpaired 3'-UA-5' at the 3' end of the minus strand that can be utilized by the mutant polymerase, and similarly, the effect of the U3A mutation is to destabilize the secondary structure, freeing 3'-terminal UA; and (iv) the N terminus of nsP4 may directly interact with the 3' terminus of the minus-strand RNA for the initiation of the plus-strand genomic RNA synthesis. This hypothesis is discussed in light of our present results as well as of previous studies of alphavirus RNAs, including defective interfering RNAs. PMID- 12551968 TI - Changes in the immunogenic properties of soluble gp140 human immunodeficiency virus envelope constructs upon partial deletion of the second hypervariable region. AB - Immunization of macaques with the soluble oligomeric gp140 form of the SF162 envelope (SF162gp140) or with an SF162gp140-derived construct lacking the central region of the V2 loop (DeltaV2gp140) results in the generation of high titers of antibodies capable of neutralizing the homologous human immunodeficiency virus type 1 (HIV-1), SF162 virus (Barnett et al. J. Virol. 75:5526-5540, 2001). However, the DeltaV2gp140 immunogen is more effective than the SF162gp140 immunogen in eliciting the generation of antibodies capable of neutralizing heterologous HIV-1 isolates. This indicates that deletion of the V2 loop alters the immunogenicity of the SF162gp140 protein. The present studies were aimed at identifying the envelope regions whose immunogenicity is altered following V2 loop deletion. We report that the antibodies elicited by the SF162gp140 immunogen recognize elements of the V1, V2, and V3 loops, the CD4-binding site, and the C1 and C2 regions on the homologous SF162 gp120. With the exception of the V1 and V2 loops, the same regions are recognized on heterologous gp120 proteins. Surprisingly, although a minority of the SF162gp140-elicited antibodies target the V3 loop on the homologous gp120, the majority of the antibodies elicited by this immunogen that are capable of binding to the heterologous gp120s tested recognize their V3 loops. Deletion of the V2 loop has two effects. First, it alters the immunogenicity of the V3 and V1 loops, and second, it renders the C5 region immunogenic. Although deletion of the V2 loop does not result in an increase in the immunogenicity of the CD4-binding site per se, the relative ratio of anti-CD4-binding site to anti-V3 loop antibodies that bind to the heterologous gp120s tested is higher in sera collected from the DeltaV2gp140-immunized animals than in the SF162gp140-immunized animals. Overall, our studies indicate that it is possible to alter the immunogenic structure of the HIV envelope by introducing specific modifications. PMID- 12551969 TI - The amino-terminal extensions of the longer Sendai virus C proteins modulate pY701-Stat1 and bulk Stat1 levels independently of interferon signaling. AB - The Sendai virus (SeV) C proteins are known to interact with Stat1 to prevent interferon (IFN)-induced pY701-Stat1 formation and IFN signaling. Nevertheless, pY701-Stat1 levels paradoxically increase during SeV infection. The C proteins also induce bulk Stat1 instability in some cells, similar to rubulavirus V proteins. We have found that SeV infection increases pY701-Stat1 levels even in cells in which bulk Stat1 levels strongly decrease. Remarkably, both the decrease in bulk Stat1 levels and the increase in pY701-Stat1 levels were found to be independent of the IFN signaling system, i.e., these events occur in mutant cells in which various components of the IFN signaling system have been disabled. Consistent with this, the C-induced decrease in Stat1 levels does not require Y701 of Stat1. We present evidence that C interacts with Stat1 in two different ways, one that prevents IFN-induced pY701-Stat1 formation and IFN signaling that has already been documented, and another that induces pY701-Stat1 formation (while decreasing bulk Stat1 levels) in a manner that does not require IFN signaling. These two types of Stat1 interaction are also distinguishable by C gene mutations. In particular, the IFN signaling-independent Stat1 interactions specifically require the amino-terminal extensions of the longer C proteins. The actions of the SeV C proteins in counteracting the cellular antiviral response are clearly more extensive than previously appreciated. PMID- 12551971 TI - Foamy virus envelope glycoprotein is sufficient for particle budding and release. AB - Foamy viruses (FVs) are classified in the family Retroviridae, but recent data have shown that they are not conventional retroviruses. Notably, several characteristics of their particle replication strategies are more similar to those of hepatitis B virus (HBV) than those of typical retroviruses. Compared to conventional retroviruses, which require only Gag proteins for budding and release of virus-like particles (VLPs), both FV and HBV require Env proteins. In the case of HBV, Env (S protein) alone is sufficient to form subviral particles (SVPs). Because FVs also depend on Env for budding, we tested whether FV Env alone could produce SVPs. The Env proteins of FV and murine leukemia virus (MuLV) were both released into cell culture supernatants and migrated into isopycnic gradients; however, unlike MuLV Env, FV Env displayed characteristics of SVPs. FV Env particles were of greater density than those of MuLV (1.11 versus 1.07 g/ml, respectively), which strongly suggested that the released proteins of FV Env were particulate. When we examined FV SVPs by immunoelectron microscopy, we found particles that were consistent in morphology, size, and staining with gold beads, similar to FV VLPs and unlike the particle-like structures of MuLV Env, which were more consistent with vesicles produced from nonspecific membrane "blebbing." Taken together, our results demonstrated that FV Env alone is sufficient for particle budding. This finding is unique among retroviruses and further demonstrated the similarities between FV and HBV. PMID- 12551970 TI - Nuclear entry of high-risk human papillomavirus type 16 E6 oncoprotein occurs via several pathways. AB - The E6 oncoprotein of high-risk human papillomavirus type 16 (HPV16) interacts with several nuclear transcription factors and coactivators in addition to cytoplasmic proteins, suggesting that nuclear import of HPV16 E6 plays a role in the cellular transformation process. In this study we have investigated the nuclear import pathways of HPV16 E6 in digitonin-permeabilized HeLa cells. We found that HPV16 E6 interacted with the karyopherin (Kap) alpha2 adapter and could enter the nucleus via a classical Kap alpha2beta1-mediated pathway. Interestingly, HPV16 E6 also interacted, via its basic nuclear localization signal (NLS) located at the C terminus, with both Kap beta1 and Kap beta2 import receptors. Binding of RanGTP to these Kap betas inhibited their interaction with HPV16 E6 NLS. In agreement with these binding data, nuclear import of the HPV16 E6 oncoprotein in digitonin-permeabilized HeLa cells could be mediated by either Kap beta1 or Kap beta2. Nuclear import via these pathways required RanGDP and was independent of GTP hydrolysis by Ran. Significantly, an E6(R124G) mutant had reduced nuclear import activity, and the E6 deletion mutant lacking (121)KKQR(124) was not imported into the nucleus. The data reveal that the HPV16 E6 oncoprotein interacts via its C-terminal NLS with several karyopherins and exploits these interactions to enter the nucleus of host cells via multiple pathways. PMID- 12551972 TI - Identification of small molecule compounds that selectively inhibit varicella zoster virus replication. AB - A series of nonnucleoside, N-alpha-methylbenzyl-N'-arylthiourea analogs were identified which demonstrated selective activity against varicella-zoster virus (VZV) but were inactive against other human herpesviruses, including herpes simplex virus. Representative compounds had potent activity against VZV early passage clinical isolates and an acyclovir-resistant isolate. Resistant viruses generated against one inhibitor were also resistant to other compounds in the series, suggesting that this group of related small molecules was acting on the same virus-specific target. Sequencing of the VZV ORF54 gene from two independently derived resistant viruses revealed mutations in ORF54 compared to the parental VZV strain Ellen sequence. Recombinant VZV in which the wild-type ORF54 sequence was replaced with the ORF54 gene from either of the resistant viruses became resistant to the series of inhibitor compounds. Treatment of VZV infected cells with the inhibitor impaired morphogenesis of capsids. Inhibitor treated cells lacked DNA-containing dense-core capsids in the nucleus, and only incomplete virions were present on the cell surface. These data suggest that the VZV-specific thiourea inhibitor series block virus replication by interfering with the function of the ORF54 protein and/or other proteins that interact with the ORF54 protein. PMID- 12551973 TI - Conserved protein kinases encoded by herpesviruses and cellular protein kinase cdc2 target the same phosphorylation site in eukaryotic elongation factor 1delta. AB - Earlier studies have shown that translation elongation factor 1delta (EF-1delta) is hyperphosphorylated in various mammalian cells infected with representative alpha-, beta-, and gammaherpesviruses and that the modification is mediated by conserved viral protein kinases encoded by herpesviruses, including UL13 of herpes simplex virus type 1 (HSV-1), UL97 of human cytomegalovirus, and BGLF4 of Epstein-Barr virus (EBV). In the present study, we attempted to identify the site in EF-1delta associated with the hyperphosphorylation by the herpesvirus protein kinases. Our results are as follows: (i) not only in infected cells but also in uninfected cells, replacement of the serine residue at position 133 (Ser-133) of EF-1delta by alanine precluded the posttranslational processing of EF-1delta, which corresponds to the hyperphosphorylation. (ii) A purified chimeric protein consisting of maltose binding protein (MBP) fused to a domain of EF-1delta containing Ser-133 (MBP-EFWt) is specifically phosphorylated in in vitro kinase assays by purified recombinant UL13 fused to glutathione S-transferase (GST) expressed in the baculovirus system. In contrast, the level of phosphorylation by the recombinant UL13 of MBP-EFWt carrying an alanine replacement of Ser-133 (MBP EFS133A) was greatly impaired. (iii) MBP-EFWt is also specifically phosphorylated in vitro by purified recombinant BGLF4 fused to GST expressed in the baculovirus system, and the level of phosphorylation of MBP-EFS133A by the recombinant BGLF4 was greatly reduced. (iv) The sequence flanking Ser-133 of EF-1delta completely matches the consensus phosphorylation site for a cellular protein kinase, cdc2, and in vitro kinase assays revealed that purified cdc2 phosphorylates Ser-133 of EF-1delta. (v) As observed with EF-1delta, the casein kinase II beta subunit (CKIIbeta) was specifically phosphorylated by UL13 in vitro, while the level of phosphorylation of CKIIbeta by UL13 was greatly diminished when a serine residue at position 209, which has been reported to be phosphorylated by cdc2, was replaced with alanine. These results indicate that the conserved protein kinases encoded by herpesviruses and a cellular protein kinase, cdc2, have the ability to target the same amino acid residues for phosphorylation. Our results raise the possibility that the viral protein kinases mimic cdc2 in infected cells. PMID- 12551974 TI - Human cytomegalovirus infection leads to accumulation of geminin and inhibition of the licensing of cellular DNA replication. AB - Previous studies have shown that infection of G(0)-synchronized human fibroblasts by human cytomegalovirus (HCMV) results in a block to cellular DNA synthesis. In this study, we have examined the effect of viral infection on the formation of the host cell DNA prereplication complex (pre-RC). We found that the Cdc6 protein level was significantly upregulated in the virus-infected cells and that there was a delay in the expression of the Mcm family of proteins. The loading of the Mcm proteins onto the DNA pre-RC complex also appeared to be defective in the virus-infected cells. This inhibition of DNA replication licensing was associated with the accumulation of geminin, a replication inhibitor. Cdt1, which participates in the loading of the Mcm proteins, was also downregulated and modified differentially in the infected cells. Early viral gene expression was sufficient for the virus-induced alteration of the pre-RC, and the immediate early protein IE1 was not required. These studies show that the inhibition of replication licensing in HCMV-infected cells is one of the multiple pathways by which the virus dysregulates the host cell cycle. PMID- 12551975 TI - The 64-kilodalton capsid protein homolog of Beet yellows virus is required for assembly of virion tails. AB - The filamentous virion of the closterovirus Beet yellows virus (BYV) consists of a long body formed by the major capsid protein (CP) and a short tail composed of the minor capsid protein (CPm) and the virus-encoded Hsp70 homolog. By using nano liquid chromatography-tandem mass spectrometry and biochemical analyses, we show here that the BYV 64-kDa protein (p64) is the fourth integral component of BYV virions. The N-terminal domain of p64 is exposed at the virion surface and is accessible to antibodies and mild trypsin digestion. In contrast, the C-terminal domain is embedded in the virion and is inaccessible to antibodies or trypsin. The C-terminal domain of p64 is shown to be homologous to CP and CPm. Mutation of the signature motifs of capsid proteins of filamentous RNA viruses in p64 results in the formation of tailless virions, which are unable to move from cell to cell. These results reveal the dual function of p64 in tail assembly and BYV motility and support the concept of the virion tail as a specialized device for BYV cell to-cell movement. PMID- 12551976 TI - Characterization of a cytolytic strain of equine infectious anemia virus. AB - A novel strain of equine infectious anemia virus (EIAV) called vMA-1c that rapidly and specifically killed infected equine fibroblasts (ED cells) but not other infectible cell lines was established. This strain was generated from an avirulent, noncytopathic strain of EIAV, MA-1. Studies with this new cytolytic strain of virus have permitted us to define viral parameters associated with EIAV induced cell killing and begin to explore the mechanism. vMA-1c infection resulted in induction of rapid cell death, enhanced fusogenic activity, and increased rates of spread in equine fibroblasts compared to other strains of EIAV. The highly cytolytic nature of vMA-1c suggested that this strain might be superinfecting equine fibroblasts. Receptor interference studies demonstrated that prior infection of equine fibroblasts with EIAV did not alter the ability of vMA-1c to infect and kill these cells. In similar studies in a canine fibroblast cell line, receptor interference did occur. vMA-1c infection of equine fibroblasts was also associated with large quantities of unintegrated viral DNA, a well-established hallmark of retroviral superinfection. Cloning of the vMA-1c genome identified nucleotide changes that would result in at least one amino acid change in all viral proteins. A chimeric infectious molecular clone containing the vMA-1c tat, S2, and env open reading frames recapitulated most of the characteristics of vMA-1c, including superinfection, fibroblast killing, and fusogenic activity. In summary, in vitro selection for a strain of EIAV that rapidly killed cells resulted in the generation of a virus that was able to superinfect these cells, presumably by the use of a novel mechanism of cell entry. This phenotype mapped to the 3' half of the genome. PMID- 12551977 TI - Mucosal priming of simian immunodeficiency virus-specific cytotoxic T-lymphocyte responses in rhesus macaques by the Salmonella type III secretion antigen delivery system. AB - Nearly all human immunodeficiency virus (HIV) infections are acquired mucosally, and the gut-associated lymphoid tissues are important sites for early virus replication. Thus, vaccine strategies designed to prime virus-specific cytotoxic T lymphocyte (CTL) responses that home to mucosal compartments may be particularly effective at preventing or containing HIV infection. The Salmonella type III secretion system has been shown to be an effective approach for stimulating mucosal CTL responses in mice. We therefore tested DeltaphoP-phoQ attenuated strains of Salmonella enterica serovar Typhimurium and S. enterica serovar Typhi expressing fragments of the simian immunodeficiency virus (SIV) Gag protein fused to the type III-secreted SopE protein for the ability to prime virus-specific CTL responses in rhesus macaques. Mamu-A*01(+) macaques were inoculated with three oral doses of recombinant Salmonella, followed by a peripheral boost with modified vaccinia virus Ankara expressing SIV Gag (MVA Gag). Transient low-level CTL responses to the Mamu-A*01 Gag(181-189) epitope were detected following each dose of Salmonella. After boosting with MVA Gag, strong Gag-specific CTL responses were consistently detected, and tetramer staining revealed the expansion of Gag(181-189)-specific CD8(+) T-cell responses in peripheral blood. A significant percentage of the Gag(181-189)-specific T-cell population in each animal also expressed the intestinal homing receptor alpha4beta7. Additionally, Gag(181-189)-specific CD8(+) T cells were detected in lymphocytes isolated from the colon. Yet, despite these responses, Salmonella primed/MVA-boosted animals did not exhibit improved control of virus replication following a rectal challenge with SIVmac239. Nevertheless, this study demonstrates the potential of mucosal priming by the Salmonella type III secretion system to direct SIV-specific cellular immune responses to the gastrointestinal mucosa in a primate model. PMID- 12551978 TI - Murine gammaherpesvirus 68 lacking thymidine kinase shows severe attenuation of lytic cycle replication in vivo but still establishes latency. AB - The lytic cycle functions of gammaherpesviruses have received relatively little attention to date, at least in part due to the lack of a convenient experimental model. The murine gammaherpesvirus 68 (MHV-68) now provides such a model and allows the roles of individual lytic cycle gammaherpesvirus proteins to be evaluated in vivo. We have used MHV-68 to determine the contribution of a gammaherpesvirus thymidine kinase (TK) to viral lytic replication and latency establishment. MHV-68 mutants with a disrupted TK gene grew normally in vitro but showed a severe attenuation of replication in the lungs after intranasal inoculation, with lytic titers at least 1,000-fold lower than those of wild-type and revertant viruses. Nevertheless, the establishment of latency by the TK deficient mutants, while delayed, was not prevented by their lytic replication deficit. The viral TK clearly plays a crucial role in the capacity of MHV-68 to replicate efficiently in its natural host but does not seem to be essential to establish a persistent infection. The potential of TK-deficient mutants as gammaherpesvirus vaccines is discussed. PMID- 12551979 TI - TT virus in the nasal secretions of children with acute respiratory diseases: relations to viremia and disease severity. AB - The natural history and pathogenic potential of the recently identified TT virus (TTV) are currently a matter of intensive investigation. In an attempt to shed some light on these issues, nasal and blood specimens of 1- to 24-month-old children hospitalized with a clinical diagnosis of acute respiratory disease (ARD) were examined for the presence, load, and genetic characteristics of TTV. The results have indicated that at least in young children, the respiratory tract not only represents a route by which abundant TTV can be shed into the environment but also may be a site of primary infection and continual replication. Although we found no compelling evidence that TTV was the direct cause of ARD in some of the children studied, the average loads of TTV were considerably higher in patients with bronchopneumonia (BP) than in those with milder ARD, raising interesting questions about the pathophysiological significance of TTV at this site. Furthermore, group 4 TTV was detected almost exclusively in children with BP. PMID- 12551980 TI - Mechanism of reduced T-cell effector functions and class-switched antibody responses to herpes simplex virus type 2 in the absence of B7 costimulation. AB - T-cell costimulation molecules B7-1 and B7-2 play an important role in activation of T cells to cytolytic effector function and production of cytokines. Interaction with B7 also causes T cells to upregulate surface molecules, such as CD40L, that effectively stimulate antibody responses in conjunction with cytokines. We have shown that mice lacking both B7-1 and B7-2 (B7KO mice), when infected intravaginally with virulent herpes simplex virus type 2 (HSV-2), developed more severe disease and higher mortality than their wild-type counterparts. We have now investigated the effects of B7 costimulation deficiency on induction of immune responses to HSV-2 infection of the genital tract. Fewer gamma interferon (IFN-gamma)-producing T cells were present in the genital lymph nodes of B7KO mice compared to wild-type mice, either acutely after primary infection or in recall responses. Less IFN-gamma and especially interleukin-10 were produced by B7KO mice, and cytolytic T-lymphocyte activity was also attenuated. Reduced expression of CD25 on CD4(+) T cells after infection of B7KO mice was consistent with deficits in T-cell activation to effector functions. Although HSV-specific immunoglobulin M (IgM) titers were comparable for both B7KO mice and wild-type mice, B7KO mice had significant deficits in HSV-specific serum IgG responses, with markedly reduced levels of IgG2a and IgG1. In addition, significantly less IgG was detected in the vaginal secretions of B7KO mice than in those from wild-type mice. CD4(+) T-cell expression of CD40L was depressed in B7KO mice in vivo and in vitro. Together with reduced cytokine production, these results suggest a mechanism for decreased IgG class switching or production. Thus, in the absence of B7 costimulation, naive T cells fail to undergo proper activation in response to HSV-2, which limits T-cell cytokine production, cytotoxic T lymphocyte activity, and provision of help for class-switched antibody responses. PMID- 12551981 TI - Baculovirus alkaline nuclease possesses a 5'-->3' exonuclease activity and associates with the DNA-binding protein LEF-3. AB - Alkaline nuclease (AN) of the Autographa californica multiple-capsid nucleopolyhedrovirus (AcMNPV) (open reading frame 133) was expressed in recombinant baculovirus as a His(6)-tagged fusion and purified by sequential chromatography on Ni-NTA-agarose, DEAE-Toyopearl, and heparin-Sepharose. At all stages of purification, AcMNPV AN was found to copurify with a 44-kDa polypeptide which was identified as the baculovirus single-stranded DNA (ssDNA)-binding (SSB) protein, LEF-3. Sedimentation analysis in glycerol gradients of highly purified samples suggested that AN and LEF-3 are associated in a complex (designated *AN/L3), predominantly as heterodimers, although oligomeric forms containing both proteins were evident. In reactions with single- or double-stranded 62-mer oligonucleotides that were labeled with (32)P at the 5' or 3' ends, *AN/L3 carried out exonucleolytic hydrolysis of both substrates exclusively in a 5'-->3' direction. Saturation of ssDNA with an excess of LEF-3 prior to the addition of *AN/L3 resulted in a marked decrease in the rate of ssDNA hydrolysis. This suggests that excess LEF-3 may protect ssDNA from digestion by a AN-LEF-3 complex, thus regulating its activity in infected cells. The association of baculovirus AN with the viral SSB LEF-3 and the 5'-->3' exonuclease activity of this complex suggests that AN and LEF-3 may participate in homologous recombination of the baculovirus genome in a manner similar to that of exonuclease (Redalpha) and DNA-binding protein (Redbeta) of the Red-mediated homologous recombination system of bacteriophage lambda. PMID- 12551983 TI - The human herpesvirus 6 U100 gene product is the third component of the gH-gL glycoprotein complex on the viral envelope. AB - The human herpesvirus 6 (HHV-6) variant A U100 gene encodes the third component of the glycoprotein H (gH)-glycoprotein L (gL)-containing complex. Glycosidase digestion analysis showed that the U100 gene products are glycoproteins consisting of an 80-kDa protein with complex N-linked oligosaccharides and a 74 kDa protein with immature, high-mannose N-linked oligosaccharides. Based on these characteristics, we designated the U100 gene products glycoprotein Q (gQ). Only the 80-kDa form of gQ was coimmunoprecipitated with an anti-gH antibody, suggesting that the 80-kDa protein associates with the gH-gL complex in HHV-6 infected cells. Furthermore, the complex was detected in purified virions, suggesting that it may play an important role in viral entry. PMID- 12551982 TI - The early expression of glycoprotein B from herpes simplex virus can be detected by antigen-specific CD8+ T cells. AB - The immune response to cutaneous herpes simplex virus type 1 (HSV-1) infection begins with remarkable rapidity. Activation of specific cytotoxic T lymphocytes (CTL) begins within hours of infection, even though the response within the draining lymph nodes peaks nearly 5 days later. HSV gene products are classified into three main groups, alpha, beta, and gamma, based on their kinetics and requirements for expression. In C57BL/6 mice, the immunodominant epitope from HSV is derived from glycoprotein B (gB(498-505)). While gB is considered a gamma or "late" gene product, previous reports have indicated that some level of gene expression may occur soon after infection. Using brefeldin A as a specific inhibitor of viral antigen presentation to major histocompatibility complex class I-restricted CTL, we have formally addressed the timing of gB peptide expression in an immunologically relevant manner following infection. Presentation of gB peptide detected by T-cell activation was first observed within 2 h of infection. Comparison with another viral epitope expressed early during infection, HSV-1 ribonucleotide reductase, demonstrated that gB is presented with the same kinetics as this classical early-gene product. Moreover, this rapidity of gB expression was further illustrated via rapid priming of naive transgenic CD8(+) T cells in vivo after HSV-1 infection of mice. These results establish that gB is expressed rapidly following HSV-1 infection, at levels capable of effectively stimulating CD8(+) T cells. PMID- 12551984 TI - Identification and molecular characterization of the RNA polymerase-binding motif of infectious bursal disease virus inner capsid protein VP3. AB - Infectious bursal disease virus (IBDV), a member of the Birnaviridae family, is the causative agent of one of the most important infectious poultry diseases. Major aspects of the molecular biology of IBDV, such as assembly and replication, are as yet poorly understood. We have previously shown that encapsidation of the putative virus-encoded RNA-dependent RNA polymerase VP1 is mediated by its interaction with the inner capsid protein VP3. Here, we report the characterization of the VP1-VP3 interaction. RNase A treatment of VP1- and VP3 containing extracts does not affect the formation of VP1-VP3 complexes, indicating that formation of the complex requires the establishment of protein protein interactions. The use of a set of VP3 deletion mutants allowed the mapping of the VP1 binding motif of VP3 within a highly charged 16-amino-acid stretch on the C terminus of VP3. This region of VP3 is sufficient to confer VP1 binding activity when fused to an unrelated protein. Furthermore, a peptide corresponding to the VP1 binding region of VP3 specifically inhibits the formation of VP1-VP3 complexes. The presence of Trojan peptides containing the VP1 binding motif in IBDV-infected cells specifically reduces infective virus production, thus showing that formation of VP1-VP3 complexes plays a critical role in IBDV replication. PMID- 12551985 TI - Lack of tumor necrosis factor alpha induces impaired proliferation of hepatitis B virus-specific cytotoxic T lymphocytes. AB - Recent studies have shown that tumor necrosis factor alpha (TNF-alpha) plays critical roles in not only viral clearance but also lymphoid tissue development and stem cell differentiation. In this study, we attempted to induce hepatitis B virus (HBV)-specific cytotoxic T lymphocytes (CTLs) by immunization of TNF-alpha knockout (TNF-alpha(-/-)) mice with HBsAg-encoding plasmid DNA. An immunization with the HBV plasmid failed to induce CTL responses in TNF-alpha(-/-) mice, although CTLs were readily induced in wild-type mice by the same protocol. Weak CTL responses were produced in TNF-alpha(-/-) mice after two sessions of immunization with the HBV plasmid; however, TNF-alpha was required to maintain the responses of these CTL lines to in vitro stimulation and, even then, the responses were lost after 3 weeks. Interestingly, a limiting dilution of a CTL line showed that HBV-specific CTL clones with high specific cytotoxicity were present in TNF-alpha(-/-) mice, but these clones again failed to proliferate for more than 3 weeks. Furthermore, since exogenously added TNF-alpha enhanced the proliferation of a TNF-alpha(-/-) clone but suppressed that of a TNF-alpha(+/+) clone in vitro, TNF-alpha also has a direct effect on the proliferation of CTLs. In conclusion, TNF-alpha is essential rather than important for the proliferation of HBV-specific CTLs both in vivo and in vitro and this effect is not only due to the activation of dendritic cells but is also induced by the direct effect on CTLs. PMID- 12551986 TI - Helicase motif Ia is involved in single-strand DNA-binding and helicase activities of the herpes simplex virus type 1 origin-binding protein, UL9. AB - UL9 is a multifunctional protein essential for herpes simplex virus type 1 (HSV 1) replication in vivo. UL9 is a member of the superfamily II helicases and exhibits helicase and origin-binding activities. It is thought that UL9 binds the origin of replication and unwinds it in the presence of ATP and the HSV-1 single stranded DNA (ssDNA)-binding protein. We have previously characterized the biochemical properties of mutants in all helicase motifs except for motif Ia (B. Marintcheva and S. Weller, J. Biol. Chem. 276:6605-6615, 2001). Structural information for other superfamily I and II helicases indicates that motif Ia is involved in ssDNA binding. By analogy, we hypothesized that UL9 motif Ia is important for the ssDNA-binding function of the protein. On the basis of sequence conservation between several UL9 homologs within the Herpesviridae family and distant homology with helicases whose structures have been solved, we designed specific mutations in motif Ia and analyzed them genetically and biochemically. Mutant proteins with residues predicted to be involved in ssDNA binding (R112A and R113A/F115A) exhibited wild-type levels of intrinsic ATPase activity and moderate to severe defects in ssDNA-stimulated ATPase activity and ssDNA binding. The S110T mutation targets a residue not predicted to contact ssDNA directly. The mutant protein with this mutation exhibited wild-type levels of intrinsic ATPase activity and near wild-type levels of ssDNA-stimulated ATPase activity and ssDNA binding. All mutant proteins lack helicase activity but were able to dimerize and bind the HSV-1 origin of replication as well as wild-type UL9. Our results indicate that residues from motif Ia contribute to the ssDNA-binding and helicase activities of UL9 and are essential for viral growth. This work represents the successful application of an approach based on a combination of bioinformatics and structural information from related proteins to deduce valuable information about a protein of interest. PMID- 12551987 TI - Distinct modes of regulation of transcription of hepatitis B virus by the nuclear receptors HNF4alpha and COUP-TF1. AB - To study the effects of the nuclear receptors (NRs) HNF4alpha and COUP-TF1 on the life cycle of hepatitis B virus (HBV), the human hepatoma cell line Huh7 was transiently cotransfected with plasmids containing the HBV genome and encoding these two NRs. Overexpression of HNF4alpha and COUP-TF1 led to a 9-fold increase and a 7- to 10-fold decrease, respectively, in viral DNA synthesis. These two NRs also exhibited distinct modes of regulation of viral transcription. Overexpression of HNF4alpha led to a more-than-10-fold increase in synthesis of the pregenomic RNA but to only a 2- to 3-fold increase in synthesis of the pre-C and S RNAs. Moreover, the NR response element within the pre-C promoter, NRRE(preC,) played the major role in activation of pregenomic RNA synthesis by HNF4alpha. On the other hand, overexpression of COUP-TF1 led to an over-10-fold repression of synthesis of both pre-C and pregenomic RNAs mediated through either NRRE(preC) or NRRE(enhI). HNF4alpha and COUP-TF1 antagonized each other's effects on synthesis of pregenomic RNA and viral DNA when they were co-overexpressed. A naturally occurring HBV variant which allows for binding by HNF4alpha but not COUP-TF1 in its NRRE(preC) exhibited significantly higher levels of synthesis of pregenomic RNA and viral DNA than wild-type HBV in coexpression experiments. Last, deletion analysis revealed that non-NRRE sequences located within both the C and pre-S1 regions are also essential for maximum activation of the pregenomic promoter by HNF4alpha but not for repression by COUP-TF1. Thus, HNF4alpha and COUP-TF1 function through different mechanisms to regulate expression of the HBV genes. PMID- 12551988 TI - Foot-and-mouth disease virus receptors: comparison of bovine alpha(V) integrin utilization by type A and O viruses. AB - Three members of the alpha(V) integrin family of cellular receptors, alpha(V)beta(1), alpha(V)beta(3), and alpha(V)beta(6), have been identified as receptors for foot-and-mouth disease virus (FMDV) in vitro. The virus interacts with these receptors via a highly conserved arginine-glycine-aspartic acid (RGD) amino acid sequence motif located within the betaG-betaH (G-H) loop of VP1. Other alpha(V) integrins, as well as several other integrins, recognize and bind to RGD motifs on their natural ligands and also may be candidate receptors for FMDV. To analyze the roles of the alpha(V) integrins from a susceptible species as viral receptors, we molecularly cloned the bovine beta(1), beta(5), and beta(6) integrin subunits. Using these subunits, along with previously cloned bovine alpha(V) and beta(3) subunits, in a transient expression assay system, we compared the efficiencies of infection mediated by alpha(V)beta(1), alpha(V)beta(3), alpha(V)beta(5), and alpha(V)beta(6) among three strains of FMDV serotype A and two strains of serotype O. While all the viruses could infect cells expressing these integrins, they exhibited different efficiencies of integrin utilization. All the type A viruses used alpha(V)beta(3) and alpha(V)beta(6) with relatively high efficiency, while only one virus utilized alpha(V)beta(1) with moderate efficiency. In contrast, both type O viruses utilized alpha(V)beta(6) and alpha(V)beta(1) with higher efficiency than alpha(V)beta(3). Only low levels of viral replication were detected in alpha(V)beta(5)-expressing cells infected with either serotype. Experiments in which the ligand-binding domains among the beta subunits were exchanged indicated that this region of the integrin subunit appears to contribute to the differences in integrin utilizations among strains. In contrast, the G-H loops of the different viruses do not appear to be involved in this phenomenon. Thus, the ability of the virus to utilize multiple integrins in vitro may be a reflection of the use of multiple receptors during the course of infection within the susceptible host. PMID- 12551989 TI - Reduction of natural adenovirus tropism to the liver by both ablation of fiber coxsackievirus and adenovirus receptor interaction and use of replaceable short fiber. AB - The initial recognition and binding of adenovirus vector to the host cell surface is mediated by interaction between the adenovirus fiber knob protein and its receptor, the coxsackievirus and adenovirus receptor (CAR). This natural tropism of adenovirus vector needs to be ablated in order to achieve targeted gene transfer. To this end, we noted that adenovirus serotype 40 (Ad40) contains two distinct long and short fibers; the short fiber is unable to recognize CAR, while the long fiber binds CAR. We generated adenovirus serotype 5-based mutants with chimeric Ad40-derived fibers, which were composed of either long or short shafts together with CAR binding or nonbinding knobs. The capacity of these adenovirus mutants for in vitro and in vivo gene transfer to liver cells was examined. In the case of primary human hepatocytes displaying a high expression level of CAR and alphav integrin, both CAR binding ability and fiber shaft length played important roles in efficient transduction. Most significantly, the high transduction efficiency observed in the liver and spleen following intravenous administration of adenovirus vector was dramatically reduced by both ablation of fiber-CAR interaction and the use of replaceable short fiber. In other tissues displaying a low level of transduction, no significant differences in transduction efficiency were observed among adenovirus vector mutants. Furthermore, incorporation of a 7-lysine-residue motif at the C-terminal end of CAR-nonbinding short fiber efficiently achieved transduction of target cells via the heparan-containing receptor. Our results demonstrated that the natural tropism of adenovirus in vivo is influenced not only by fiber-CAR interaction but also by fiber shaft length. Furthermore, our strategy may be useful for retargeting adenovirus to particular tumors and tissue types with specific receptors. PMID- 12551990 TI - Effective vaccination against long-term gammaherpesvirus latency. AB - The fundamental question of whether a primed immune system is capable of preventing latent gammaherpesvirus infection remains unanswered. Recent studies showing that vaccination can reduce acute replication and short-term latency but cannot alter long-term latency further call into question the possibility of achieving sterilizing immunity against gammaherpesviruses. Using the murine gammaherpesvirus 68 (gammaHV68) system, we demonstrate that it is possible to effectively vaccinate against long-term latency. By immunizing mice with a gammaHV68 mutant virus that is deficient in its ability to reactivate from latency, we reduced latent infection of wild-type challenge virus to a level below the limit of detection. Establishment of latency was inhibited by vaccination regardless of whether mice were challenged intraperitoneally or intranasally. Passive transfer of antibody from vaccinated mice could partially reconstitute the effect, demonstrating that antibody is an important component of vaccination. These results demonstrate the potential of a memory immune response against gammaherpesviruses to alter long-term latency and suggest that limiting long-term latent infection in a clinically relevant situation is an attainable goal. PMID- 12551991 TI - Identification of a receptor-binding domain of the spike glycoprotein of human coronavirus HCoV-229E. AB - Human coronavirus HCoV-229E uses human aminopeptidase N (hAPN) as its receptor (C. L. Yeager et al., Nature 357:420-422, 1992). To identify the receptor-binding domain of the viral spike glycoprotein (S), we expressed soluble truncated histidine-tagged S glycoproteins by using baculovirus expression vectors. Truncated S proteins purified by nickel affinity chromatography were shown to be glycosylated and to react with polyclonal anti-HCoV-229E antibodies and monoclonal antibodies to the viral S protein. A truncated protein (S(547)) that contains the N-terminal 547 amino acids bound to 3T3 mouse cells that express hAPN but not to mouse 3T3 cells transfected with empty vector. Binding of S(547) to hAPN was blocked by an anti-hAPN monoclonal antibody that inhibits binding of virus to hAPN and blocks virus infection of human cells and was also blocked by polyclonal anti-HCoV-229E antibody. S proteins that contain the N-terminal 268 or 417 amino acids did not bind to hAPN-3T3 cells. Antibody to the region from amino acid 417 to the C terminus of S blocked binding of S(547) to hAPN-3T3 cells. Thus, the data suggest that the domain of the spike protein between amino acids 417 and 547 is required for the binding of HCoV-229E to its hAPN receptor. PMID- 12551992 TI - Phosphatidylinositol 3-kinase regulates human immunodeficiency virus type 1 replication following viral entry in primary CD4+ T lymphocytes and macrophages. AB - Human immunodeficiency virus (HIV) gp120 induces multiple cellular signaling pathways, including the phosphatidylinositol 3-kinase (PI3-kinase) pathway. The role of the PI3-kinase pathway in HIV-1 replication is not understood. Here we examined whether HIV-1 gp120 upregulates the PI3-kinase pathway and whether PI3 kinase activity plays a role in virus replication in primary human CD4(+) T cells and macrophages. Soluble and virion-associated HIV-1 gp120 induced calcium mobilization and phosphorylation of the PI3-kinase downstream effectors PKB/Akt and p70 S6 kinase. gp120-induced PI3-kinase activity and calcium mobilization were inhibited by pertussis toxin and blocking antibodies directed against CCR5 and CXCR4, suggesting that the signaling is mediated through the chemokine receptor. The PI3-kinase inhibitor LY294002 inhibited infection of CD4(+) T cells and macrophages with X4 and R5 HIV-1-pseudotyped viruses at concentrations that did not induce cell toxicity or downregulate HIV-1 coreceptor expression. When gp120-induced signaling was bypassed with the vesicular stomatitis virus G envelope protein, infection was still sensitive to PI3-kinase inhibition, suggesting that basal PI3-kinase activity is required for infection. LY294002 inhibited HIV-1 infection when added after viral entry and did not affect formation of the HIV-1 reverse transcriptase products R/U5 and long terminal repeat/Gag in the presence of the inhibitor. However, when the inhibitor was added after viral integration had occurred, no inhibition of HIV infection was observed. Our studies show that inhibition of the PI3-kinase signaling pathway suppresses virus infection post-viral entry and post-reverse transcription but prior to HIV gene expression. This type of host-virus interaction has implications for anti-HIV therapeutics that target cellular signaling machinery. PMID- 12551993 TI - G protein-dependent CCR5 signaling is not required for efficient infection of primary T lymphocytes and macrophages by R5 human immunodeficiency virus type 1 isolates. AB - The requirement of human immunodeficiency virus (HIV)-induced CCR5 activation for infection by R5 HIV type 1 (HIV-1) strains remains controversial. Ectopic CCR5 expression in CD4(+)-transformed cells or pharmacological inhibition of G(alpha)i proteins coupled to CCR5 left unsolved whether CCR5-dependent cell activation is necessary for the HIV life cycle. In this study, we investigated the role played by HIV-induced CCR5-dependent cell signaling during infection of primary CD4 expressing leukocytes. Using lentiviral vectors, we restored CCR5 expression in T lymphocytes and macrophages from individuals carrying the homozygous 32-bp deletion of the CCR5 gene (ccr5 Delta32/Delta32). Expression of wild-type (wt) CCR5 in ccr5 Delta32/Delta32 cells permitted infection by R5 HIV isolates. We assessed the capacity of a CCR5 derivative carrying a mutated DRY motif (CCR5 R126N) in the second intracellular loop to work as an HIV-1 coreceptor. The R126N mutation is known to disable G protein coupling and agonist-induced signal transduction through CCR5 and other G protein-coupled receptors. Despite its inability to promote either intracellular calcium mobilization or cell chemotaxis, the inactive CCR5-R126N mutant provided full coreceptor function to several R5 HIV-1 isolates in primary cells as efficiently as wt CCR5. We conclude that in a primary, CCR5-reconstituted CD4(+) cell environment, G protein signaling is dispensable for R5 HIV-1 isolates to actively infect primary CD4(+) T lymphocytes or macrophages. PMID- 12551994 TI - The coxsackie B virus and adenovirus receptor resides in a distinct membrane microdomain. AB - The coxsackie B virus and adenovirus receptor (CAR) is a member of the immunoglobulin superfamily. In addition to activity as a viral receptor, it may play a role in cellular adhesion. We asked what determines the cell membrane microdomain of CAR. We found that CAR is localized to a novel lipid-rich microdomain similar to that of the low-density lipoprotein receptor (LDLR) but distinct from that of a CAR variant that exhibited traditional lipid raft localization via fusion to a glycosylphosphatidylinositol (GPI) tail. The cytoplasmic tail determines its membrane localization, since deletion of this domain resulted in mislocalization. Results indicate that CAR, CAR-LDLR, and LDLR reside in a novel lipid raft that is distinct from caveolin-1-containing caveolae and GPI-linked proteins. Residence in a lipid-rich domain provides a mechanism that allows CAR to interact with other cell adhesion proteins and yet function as an adenovirus receptor. PMID- 12551995 TI - An alternate pathway for recruiting template RNA to the brome mosaic virus RNA replication complex. AB - The multidomain RNA replication protein 1a of brome mosaic virus (BMV), a positive-strand RNA virus in the alphavirus-like superfamily, plays key roles in assembly and function of the viral RNA replication complex. 1a, which encodes RNA capping and helicase-like domains, localizes to endoplasmic reticulum membranes, recruits BMV 2a polymerase and viral RNA templates, and forms membrane-bound, capsid-like spherules in which RNA replication occurs. cis-acting signals necessary and sufficient for RNA recruitment by 1a have been mapped in BMV genomic RNA2 and RNA3. Both signals comprise an extended stem-loop whose apex matches the conserved sequence and structure of the TPsiC stem-loop in tRNAs (box B). Mutations show that this box B motif is crucial to 1a responsiveness of wild type RNA2 and RNA3. We report here that, unexpectedly, some chimeric mRNAs expressing the 2a polymerase open reading frame from RNA2 were recruited by 1a to the replication complex and served as templates for negative-strand RNA synthesis, despite lacking the normally essential, box B-containing 5' signal. Further studies showed that this template recruitment required high-efficiency translation of the RNA templates. Moreover, multiple small frameshifting insertion or deletion mutations throughout the N-terminal region of the open reading frame inhibited this template recruitment, while an in-frame insertion did not. Providing 2a in trans did not restore template recruitment of RNAs with frameshift mutations. Only those deletions in the N-terminal region of 2a that abolished 2a interaction with 1a abolished template recruitment of the RNA. These and other results indicate that this alternate pathway for 1a-dependent RNA recruitment involves 1a interaction with the translating mRNA via the 1a interactive N-terminal region of the nascent 2a polypeptide. Interaction with nascent 2a also may be involved in 1a recruitment of 2a polymerase to membranes. PMID- 12551996 TI - B cells and antibody play critical roles in the immediate defense of disseminated infection by West Nile encephalitis virus. AB - West Nile virus (WNV) causes severe central nervous system (CNS) infection primarily in humans who are immunocompromised or elderly. In this study, we addressed the mechanism by which the immune system limits dissemination of WNV infection by infecting wild-type and immunodeficient inbred C57BL/6J mice with a low-passage WNV isolate from the recent epidemic in New York state. Wild-type mice replicated virus extraneuronally in the draining lymph nodes and spleen during the first 4 days of infection. Subsequently, virus spread to the spinal cord and the brain at virtually the same time. Congenic mice that were genetically deficient in B cells and antibody (microMT mice) developed increased CNS viral burdens and were vulnerable to lethal infection at low doses of virus. Notably, an approximately 500-fold difference in serum viral load was detected in micro MT mice as early as 4 days after infection, a point in the infection when low levels of neutralizing immunoglobulin M antibody were detected in wild-type mice. Passive transfer of heat-inactivated serum from infected and immune wild type mice protected micro MT mice against morbidity and mortality. We conclude that antibodies and B cells play a critical early role in the defense against disseminated infection by WNV. PMID- 12551998 TI - A functional epitope determinant on domain III of the Japanese encephalitis virus envelope protein interacted with neutralizing-antibody combining sites. AB - The envelope (E) protein of Japanese encephalitis virus (JEV) is associated with viral binding to cellular receptors, membrane fusion, and the induction of protective neutralizing-antibody responses in hosts. Most previous studies have not provided detailed molecular information about the spatial configuration of the functional epitopes on domain III of the E protein. Here site-directed mutagenesis was performed to demonstrate that the functional epitope determinants at Ser331 and Asp332 on domain III of the JEV E protein interacted with neutralizing monoclonal antibody (MAb) E3.3. Bacterial expression of the recombinant Fab E3.3 confirmed the molecular interactions of Arg94 in complementary determining region H3 with Ser331 and Asp332 on domain III. This study elucidates the detailed molecular structures of the neutralizing epitope determinants on JEV domain III, which can provide useful information for designing new vaccines. PMID- 12551997 TI - Molecular characteristics of human immunodeficiency virus type 1 subtype C viruses from KwaZulu-Natal, South Africa: implications for vaccine and antiretroviral control strategies. AB - The KwaZulu-Natal region of South Africa is experiencing an explosive outbreak of human immunodeficiency virus type 1 (HIV-1) subtype C infections. Understanding the genetic diversity of C viruses and the biological consequences of this diversity is important for the design of effective control strategies. We analyzed the protease gene, the first 935 nucleotides of reverse transcriptase, and the C2V5 envelope region of a representative set of 72 treatment-naive patients from KwaZulu-Natal and correlated the results with amino acid signature and resistance patterns. Phylogenetic analysis revealed multiple clusters or "lineages" of HIV-1 subtype C that segregated with other C viruses from southern Africa. The same pattern was observed for both black and Indian subgroups and for retrospective specimens collected prior to 1990, indicating that multiple sublineages of HIV-1 C have been present in KwaZulu-Natal since the early stages of the epidemic. With the exception of three nonnucleoside reverse transcriptase inhibitor mutations, no primary resistance mutations were identified. Numerous accessory polymorphisms were present in the protease, but none were located at drug-binding or active sites of the enzyme. One frequent polymorphism, I93L, was located near the protease/reverse transcriptase cleavage site. In the envelope, disruption of the glycosylation motif at the beginning of V3 was associated with the presence of an extra protein kinase C phosphorylation site at codon 11. Many polymorphisms were embedded within cytotoxic T lymphocyte or overlapping cytotoxic T-lymphocyte/T-helper epitopes, as defined for subtype B. This work forms a baseline for future studies aimed at understanding the impact of genetic diversity on vaccine efficacy and on natural susceptibility to antiretroviral drugs. PMID- 12551999 TI - Pseudotyped lentivirus vectors derived from simian immunodeficiency virus SIVagm with envelope glycoproteins from paramyxovirus. AB - We describe the development of novel lentivirus vectors based on simian immunodeficiency virus from African green monkey (SIVagm) pseudotyped with Sendai virus (SeV) envelope glycoproteins. SeV fusion (F) and hemagglutinin neuraminidase (HN) proteins were successfully incorporated into the SIVagm-based vector by truncation of the cytoplasmic tail of the F protein and by addition of the cytoplasmic tail of SIVagm transmembrane envelope protein to the N terminus of the HN protein. As with the vesicular stomatitis virus G glycoprotein pseudotyped vector, the mutant SeV F- and HN-pseudotyped SIVagm vector was able to transduce various types of animal and human cell lines. Furthermore, the vector was able to transduce an enhanced green fluorescent protein reporter gene into polarized epithelial cells of rat trachea from the apical and basolateral sides. Therefore, SeV F- and HN-pseudotyped SIVagm vectors have considerable potential for effective use in gene therapy for various therapies, including respiratory diseases. PMID- 12552000 TI - Cell penetration and trafficking of polyomavirus. AB - The murine polyomavirus (Py) enters mouse fibroblasts and kidney epithelial cells via an endocytic pathway that is caveola-independent (as well as clathrin independent). In contrast, uptake of simian virus 40 into the same cells is dependent on caveola. Following the initial uptake of Py, both microtubules and microfilaments play roles in trafficking of the virus to the nucleus. Colcemid, which disrupts microtubules, inhibits the ability of Py to reach the nucleus and replicate. Paclitaxel, which stabilizes microtubules and prevents microtubule turnover, has no effect, indicating that intact but not dynamic microtubules are required for Py infectivity. Compounds that disrupt actin filaments enhance Py uptake while stabilization of actin filaments impedes Py infection. Virus particles are seen in association with actin in cells treated with microfilament disrupting or filament-stabilizing agents at levels comparable to those in untreated cells, suggesting that a dynamic state of the microfilament system is important for Py infectivity. PMID- 12552001 TI - Molluscum contagiosum virus interleukin-18 (IL-18) binding protein is secreted as a full-length form that binds cell surface glycosaminoglycans through the C terminal tail and a furin-cleaved form with only the IL-18 binding domain. AB - Some poxviruses and their mammalian hosts encode homologous proteins that bind interleukin-18 (IL-18) with high affinity and inhibit IL-18-mediated immune responses. MC54L, the IL-18 binding protein of the human poxvirus that causes molluscum contagiosum, is unique in having a C-terminal tail of nearly 100 amino acids that is dispensable for IL-18 binding. When recombinant MC54L was expressed and purified via a C-terminal six-histidine tag, a shorter fragment was detected in addition to the full-length protein. This C-terminal fragment resulted from the cleavage of MC54L by cellular furin, as it was greatly diminished when furin was specifically inhibited or when a furin-deficient cell line was used for expression. Furthermore, the N- and C-terminal fragments of MC54L were generated by cleavage of the recombinant protein with furin in vitro. The furin cleavage site was mapped within a 32-amino-acid segment that is C terminal to the IL-18 binding domain. Full-length MC54L, but not the N-terminal IL-18 binding fragment, bound to cells and to purified heparin and other glycosaminoglycans that are commonly found on the cell surface and in the extracellular matrix. MC54L bound to heparin with a nanomolar K(d) and could simultaneously bind to IL-18. Their different glycosaminoglycan and cell binding properties may allow the long and short forms of MC54L to inactivate IL-18 near the site of infection and at more distal locations, respectively. PMID- 12552002 TI - Kaposi's sarcoma-like tumors in a human herpesvirus 8 ORF74 transgenic mouse. AB - The product of human herpesvirus 8 (HHV-8) open reading frame 74 (ORF74) is related structurally and functionally to cellular chemokine receptors. ORF74 activates several cellular signaling pathways in the absence of added ligands, and NIH 3T3 cells expressing ORF74 are tumorigenic in nude mice. We have generated a line of transgenic (Tg) mice with ORF74 driven by the simian virus 40 early promoter. A minority (approximately 30%) of the Tg mice, including the founder, developed tumors resembling Kaposi's sarcoma (KS) lesions, which occurred most typically on the tail or legs. The tumors were highly vascularized, had a spindle cell component, expressed VEGF-C mRNA, and contained a majority of CD31(+) cells. CD31 and VEGF-C are typically expressed in KS. Tumors generally (but not always) occurred at single sites and most were relatively indolent, although several mice developed large visceral tumors. ORF74 was expressed in a minority of cells in the Tg tumors and in a few other tissues of mice with tumors; mice without tumors did not express detectable ORF74 in any tissues tested. Cell lines established from tumors expressed ORF74 in a majority of cells, expressed VEGF-C mRNA, and were tumorigenic in nude mice. The resultant tumors grew rapidly, metastasized, and continued to express ORF74. Cell lines established from these secondary tumors also expressed ORF74 and were tumorigenic. These data strongly suggest that ORF74 plays a role in the pathology of KS and confirm and extend previous findings on the tumorigenic potential of ORF74. PMID- 12552003 TI - Developing novel oncolytic adenoviruses through bioselection. AB - Mutants of human adenovirus 5 (Ad5) with enhanced oncolytic activity were isolated by using a procedure termed bioselection. Two mutants, ONYX-201 and ONYX 203, were plaque purified from a pool of randomly mutagenized Ad5 that was repeatedly passaged in the human colorectal cancer cell line HT29, and they were subsequently characterized. ONYX-201 and ONYX-203 replicated more rapidly in HT29 cells than wild-type Ad5, and they lysed HT29 cells up to 1,000-fold more efficiently. The difference was most profound when cells were infected at a relatively low multiplicity of infection, presumably due to the compounding effects of multiple rounds of infection. This enhanced cytolytic activity was observed not only in HT29 cells but also in many other human cancer cell lines tested. In contrast, the cytotoxicity of the bioselected mutants in a number of normal primary human cells was similar to that of wild-type Ad5, thus enhancing the therapeutic index (cytotoxicity in tumor cells versus that in normal cells) of these oncolytic agents. Both ONYX-201 and -203 contain seven single-base-pair mutations when compared with Ad5, four of which were common between ONYX-201 and 203. The mutation at nucleotide 8350, shared by both mutant viruses, was shown to be essential for the observed phenotypes. This mutation was mapped to the i leader region of the major late transcription unit, resulting in the truncation of 21 amino acids from the C terminus of the i-leader protein. This work demonstrates that bioselection is a powerful tool for developing novel tumor selective oncolytic viruses. Other potential applications of this technology are discussed. PMID- 12552004 TI - E1A sensitizes cells to tumor necrosis factor alpha by downregulating c-FLIP S. AB - Tumor necrosis factor alpha (TNF-alpha) activates both apoptosis and NF-kappaB dependent survival pathways, the former of which requires inhibition of gene expression to be manifested. c-FLIP is a TNF-alpha-induced gene that inhibits caspase-8 activation during TNF-alpha signaling. Adenovirus infection and E1A expression sensitize cells to TNF-alpha by allowing apoptosis in the absence of inhibitors of gene expression, suggesting that it may be disabling a survival signaling pathway. E1A promoted TNF-alpha-mediated activation of caspase-8, suggesting that sensitivity was occurring at the level of the death-inducing signaling complex. Furthermore, E1A expression downregulated c-FLIP(S) expression and prevented its induction by TNF-alpha. c-FLIP(S) and viral FLIP expression rescued E1A-mediated sensitization to TNF-alpha by restoring the resistance of caspase-8 to activation, thereby preventing cell death. E1A inhibited TNF-alpha dependent induction of c-FLIP(S) mRNA and stimulated ubiquitination- and proteasome-dependent degradation of c-FLIP(S) protein. Since elevated c-FLIP levels confer resistance to apoptosis and promote tumorigenicity, interference with its induction by NF-kappaB and stimulation of its destruction in the proteasome may provide novel therapeutic approaches for facilitating the elimination of apoptosis-refractory tumor cells. PMID- 12552005 TI - Early induction and maintenance of Env-specific T-helper cells following human immunodeficiency virus type 1 infection. AB - Mounting evidence points to a role for CD4+ T-helper (Th) cell activities in controlling human immunodeficiency virus type 1 (HIV-1) infection. To determine the induction and evolution of Th responses following acute infection, we prospectively analyzed Env- and Gag-specific Th responses longitudinally for 92 patients with acute (n = 28) or early (n = 64) HIV-1 infection (median, 55 days postinfection [DPI]). The probability of detecting HIV-1-specific lymphoproliferative responses was remarkably low, and when present, the responses were more likely to be Gag specific than Env specific (16 versus 5%). Env specific responses were significantly more common in patients presenting at <30 DPI than in those presenting at 30 to 365 DPI (21 versus 0.5%, P = 0.001). By contrast, Gag-specific responses occurred with similar frequencies among subjects presenting at <30 DPI and 30 to 365 DPI (13 versus 17%, P = 0.6). After treatment, and regardless of the duration of infection before therapy, Gag specific Th responses predominated. Furthermore, some acutely infected subjects lost detectable Env-specific Th proliferative responses, which failed to reemerge upon treatment. Detailed analysis for one such subject revealed Env-specific lymphoproliferation at 11 DPI but no detectable Env-specific lymphoproliferation or ex vivo gamma interferon (IFN-gamma) secretion at multiple subsequent time points. Env-specific CD4+ T-cell clones from 11 DPI recognized six epitopes in both conserved and variable regions within gp120 and gp41, exhibited major histocompatibility complex-restricted cytotoxicity, and secreted high levels of antiviral cytokines. T-cell receptor clonal transcript analyses and autologous virus sequencing revealed that Th cells induced during acute infection were maintained and there were no Th escape mutations. Subsequent analysis for this subject and six of seven others revealed detectable IFN-gamma-secreting cells, but only following in vitro gp160 stimulation. In summary, we conclude that Env specific Th responses are elicited very early in acute infection and may precede Gag-specific responses. The inability to detect Env-specific Th responses over time and despite antiretroviral therapy may reflect low frequencies and impaired proliferative capacity, and viral escape is not necessary for this to occur. PMID- 12552007 TI - Interaction of the equine herpesvirus 1 EICP0 protein with the immediate-early (IE) protein, TFIIB, and TBP may mediate the antagonism between the IE and EICP0 proteins. AB - The equine herpesvirus 1 (EHV-1) immediate-early (IE) and EICP0 proteins are potent trans-activators of EHV-1 promoters; however, in transient-transfection assays, the IE protein inhibits the trans-activation function of the EICP0 protein. Assays with IE mutant proteins revealed that its DNA-binding domain, TFIIB-binding domain, and nuclear localization signal may be important for the antagonism between the IE and EICP0 proteins. In vitro interaction assays with the purified IE and EICP0 proteins indicated that these proteins interact directly. At late times postinfection, the IE and EICP0 proteins colocalized in the nuclei of infected equine cells. Transient-transfection assays showed that the EICP0 protein trans-activated EHV-1 promoters harboring only a minimal promoter region (TATA box and cap site), suggesting that the EICP0 protein trans activates EHV-1 promoters by interactions with general transcription factor(s). In vitro interaction assays revealed that the EICP0 protein interacted directly with the basal transcription factors TFIIB and TBP and that the EICP0 protein (amino acids [aa] 143 to 278) mediated the interaction with aa 125 to 174 of TFIIB. Our unpublished data showed that the IE protein interacts with the same domain (aa 125 to 174) of TFIIB and with TBP. Taken together, these results suggested that interaction of the EICP0 protein with the IE protein, TFIIB, and TBP may mediate the antagonism between the IE and EICP0 proteins. PMID- 12552008 TI - Herpes simplex virus gE/gI expressed in epithelial cells interferes with cell-to cell spread. AB - The herpes simplex virus (HSV) glycoprotein heterodimer gE/gI plays an important role in virus cell-to-cell spread in epithelial and neuronal tissues. In an analogous fashion, gE/gI promotes virus spread between certain cell types in culture, e.g., keratinocytes and epithelial cells, cells that are polarized or that form extensive cell junctions. One mechanism by which gE/gI facilitates cell to-cell spread involves selective sorting of nascent virions to cell junctions, a process that requires the cytoplasmic domain of gE. However, the large extracellular domains of gE/gI also appear to be involved in cell-to-cell spread. Here, we show that coexpression of a truncated form of gE and gI in a human keratinocyte line, HaCaT cells, decreased the spread of HSV between cells. This truncated gE/gI was found extensively at cell junctions. Expression of wild-type gE/gI that accumulates at intracellular sites, in the trans-Golgi network, did not reduce cell-to-cell spread. There was no obvious reduction in production of infectious HSV in cells expressing gE/gI, and virus particles accumulated at cell junctions, not at intracellular sites. Expression of HSV gD, which is known to bind virus receptors, also blocked cell-to-cell spread. Therefore, like gD, gE/gI appears to be able to interact with cellular components of cell junctions, gE/gI receptors which can promote HSV cell-to-cell spread. PMID- 12552009 TI - PrP polymorphisms tightly control sheep prion replication in cultured cells. AB - Prion diseases are fatal neurodegenerative disorders of animals and humans that are characterized by the conversion of the host-encoded prion protein (PrP) to an abnormal isoform. In several species, including humans, polymorphisms in the gene encoding the PrP protein tightly control susceptibility of individuals toward this disease. In the present study we show that Rov cells expressing an ovine PrP allele ((VRQ)PrP) associated with high susceptibility of sheep to scrapie were very sensitive to sheep prion transmission and replicated the agent to high titers. In contrast, we did not find any evidence of infection when Rov cells expressed similar levels of a PrP variant ((ARR)PrP) linked to resistance. Our data provide the first direct evidence that natural PrP polymorphisms may affect prion susceptibility by controlling prion replication at the cell level. The study of how PrP polymorphisms influence the genetic control of prion propagation in cultured Rov cells may help elucidate basic mechanisms of prion replication. PMID- 12552011 TI - Amino acid substitutions in VP2 residues contacting sialic acid in low neurovirulence BeAn virus dramatically reduce viral binding and spread of infection. AB - Theiler's murine encephalomyelitis viruses (TMEV) consist of two groups, the high and low-neurovirulence groups, based on lethality in intracerebrally inoculated mice. Low-neurovirulence TMEV result in a persistent central nervous system infection in mice, leading to an inflammatory demyelinating pathology and disease. Low- but not high-neurovirulence strains use sialic acid as an attachment factor. The recent resolution of the crystal structure of the low neurovirulence DA virus in complex with the sialic acid mimic sialyllactose demonstrated that four capsid residues make contact with sialic acid through noncovalent hydrogen bonds. To systematically test the importance of these sialic acid-binding residues in viral entry and infection, we mutated three VP2 puff B amino acids proposed to make contact with sialic acid and analyzed the consequences of each amino acid substitution on viral entry and spread. The fourth residue is in the VP3-VP1 cleavage dipeptide and could not be mutated. Our data suggest that residues Q2161 and G2174 are directly involved in BeAn virus attachment to sialic acid and that substitutions of these two residues result in the loss of or reduced viral binding and hemagglutination and in the inability to spread among BHK-21 cells. In addition, a gain of function-revertant virus was recovered with the Q2161A mutation after prolonged passage in cells. PMID- 12552010 TI - High mutant frequency in populations of a DNA virus allows evasion from antibody therapy in an immunodeficient host. AB - The degree of genetic heterogeneity of DNA virus populations in nature and its consequences for disease control are virtually unknown. The parvovirus minute virus of mice (MVMi) was used here to investigate (i) the frequency of antibody escape mutants in populations of a DNA virus and (ii) the ability of a DNA virus to evade in the long-term a passive monoclonal antibody (MAb) therapy in an immunodeficient natural host. Independent clonal populations of MVMi harbored a high proportion of mutants resistant to neutralizing MAb (mutant frequency = [2.8 +/- 0.5] x 10(-5)) that rapidly evolved under antibody pressure in culture to become mixtures dominated by genotypically diverse escape mutants. Immunodeficient mice naturally infected with clonal populations of MVMi and subsequently treated by intravenous injections of MAb were initially protected from the characteristic viral induced lethal leukopenia. However, some treated animals developed a delayed severe leukopenic syndrome associated with the emergence of genetically heterogeneous populations of MAb-resistant mutants in the MVMi main target organs. The 11 plaque-purified viruses analyzed from an antibody-resistant population obtained from one animal corresponded to four different mutant genotypes, although their consensus sequence remained wild type. All cloned escape mutants harbored single radical amino acid changes within a stretch of seven residues in a surface-exposed loop at the threefold axes of the capsid. This antigenic site, which can tolerate radical changes preserving MVMi pathogenic potential, may thereby allow the virus to evade the immune control. These findings indicate a high genetic heterogeneity and rapid adaptation of populations of a mammal DNA virus in vivo and provide a genetic basis for the failure of passive immunotherapy in the natural host. PMID- 12552012 TI - Structure and mechanism of a coreceptor for infection by a pathogenic feline retrovirus. AB - Infection of T lymphocytes by the cytopathic retrovirus feline leukemia virus subgroup T (FeLV-T) requires FeLIX, a cellular coreceptor that is encoded by an endogenous provirus and closely resembles the receptor-binding domain (RBD) of feline leukemia virus subgroup B (FeLV-B). We determined the structure of FeLV-B RBD, which has FeLIX activity, to a 2.5-A resolution by X-ray crystallography. The structure of the receptor-specific subdomain of this glycoprotein differs dramatically from that of Friend murine leukemia virus (Fr-MLV), which binds a different cell surface receptor. Remarkably, we find that Fr-MLV RBD also activates FeLV-T infection of cells expressing the Fr-MLV receptor and that FeLV B RBD is a competitive inhibitor of infection under these conditions. These studies suggest that FeLV-T infection relies on the following property of mammalian leukemia virus RBDs: the ability to couple interaction with one of a variety of receptors to the activation of a conserved membrane fusion mechanism. A comparison of the FeLV-B and Fr-MLV RBD structures illustrates how receptor specific regions are linked to conserved elements critical for postbinding events in virus entry. PMID- 12552013 TI - The interaction between the major capsid protein and the smallest capsid protein of human cytomegalovirus is dependent on two linear sequences in the smallest capsid protein. AB - The assembly of human cytomegalovirus (HCMV) with recombinant systems has not been accomplished. An understanding of specific interactions between individual capsid proteins could point to unique characteristics of the assembly process of HCMV capsids. Similar to its herpes simplex virus counterpart, VP26 (UL35), the HCMV smallest capsid protein (SCP; UL48/49) decorates hexons in the mature capsid. In contrast to VP26, the HCMV SCP is essential for virus assembly. In this study we have shown that the major capsid protein (MCP) and the SCP interact in the cytoplasm of transfected cells and can be coprecipitated from insect cells expressing the MCP and the SCP. Using a two-hybrid reporter assay, we demonstrated that two linear sequences within the SCP are sufficient for SCP and MCP interactions. PMID- 12552014 TI - Expression of the major histocompatibility complex class I molecule Mamu-A*01 is associated with control of simian immunodeficiency virus SIVmac239 replication. AB - Several HLA alleles are associated with attenuated human immunodeficiency virus disease progression. We explored the relationship between the expression of particular major histocompatibility complex (MHC) class I alleles and viremia in simian immunodeficiency virus SIV(mac)239-infected macaques. Of the common MHC class I alleles, animals that expressed Mamu-A*01 exhibited the best control of viral replication. PMID- 12552015 TI - Adeno-associated virus type 2-mediated gene transfer: role of cellular T-cell protein tyrosine phosphatase in transgene expression in established cell lines in vitro and transgenic mice in vivo. AB - The use of adeno-associated virus type 2 (AAV) vectors has gained attention as a potentially useful alternative to the more commonly used retrovirus and adenovirus vectors for human gene therapy. However, the transduction efficiency of AAV vectors varies greatly in different cells and tissues in vitro and in vivo. We have documented that a cellular protein that binds the immunosuppressant drug FK506, termed the FK506-binding protein (FKBP52), interacts with the single stranded D sequence within the AAV inverted terminal repeats, inhibits viral second-strand DNA synthesis, and consequently limits high-efficiency transgene expression (K. Qing, J. Hansen, K. A. Weigel-Kelley, M. Tan, S. Zhou, and A. Srivastava, J. Virol., 75: 8968-8976, 2001). FKBP52 can be phosphorylated at both tyrosine and serine/threonine residues, but only the phosphorylated forms of FKBP52 interact with the D sequence. Furthermore, the tyrosine-phosphorylated FKBP52 inhibits AAV second-strand DNA synthesis by greater than 90%, and the serine/threonine-phosphorylated FKBP52 causes approximately 40% inhibition, whereas the dephosphorylated FKBP52 has no effect on AAV second-strand DNA synthesis. In the present study, we have identified that the tyrosine phosphorylated form of FKBP52 is a substrate for the cellular T-cell protein tyrosine phosphatase (TC-PTP). Deliberate overexpression of the murine wild-type (wt) TC-PTP gene, but not that of a cysteine-to-serine (C-S) mutant, caused tyrosine dephosphorylation of FKBP52, leading to efficient viral second-strand DNA synthesis and resulting in a significant increase in AAV-mediated transduction efficiency in HeLa cells in vitro. Both wt and C-S mutant TC-PTP expression cassettes were also used to generate transgenic mice. Primitive hematopoietic stem/progenitor cells from wt TC-PTP-transgenic mice, but not from C-S mutant TC-PTP-transgenic mice, could be successfully transduced by recombinant AAV vectors. These studies corroborate the fact that tyrosine phosphorylation of the cellular FKBP52 protein strongly influences AAV transduction efficiency, which may have important implications in the optimal use of AAV vectors in human gene therapy. PMID- 12552016 TI - Novel gene product of Thogoto virus segment 6 codes for an interferon antagonist. AB - Thogoto virus (THOV) is a tick-transmitted orthomyxovirus with a genome of six negative-stranded RNA segments. The sixth segment encodes two different transcripts: a spliced transcript that is translated into the matrix protein (M) and an unspliced transcript. Here, we report that the unspliced transcript encodes an elongated form of M named ML. A THOV isolate deficient in ML expression was an efficient interferon inducer, whereas ML-expressing wild-type strains were poor interferon inducers. These results were confirmed with recombinant THOVs rescued from cDNAs. Expression of ML efficiently suppressed activation of the beta interferon promoter by double-stranded RNA. These results indicate that ML is an accessory protein that functions as a potent interferon antagonist by blocking transcriptional activation of alpha/beta interferons. PMID- 12552017 TI - Targeted retroviral infection of tumor cells by receptor cooperation. AB - Retroviruses expressing two different receptor-binding domains linked by proline rich spacers infect only cells expressing both retroviral receptors (Valsesia Wittman et al., EMBO J. 6:1214-1223, 1997). Here we apply this receptor cooperation strategy to target human tumor cells by linking single-chain antibodies recognizing tumor antigens via proline-rich spacers to the 4070A murine leukemia virus surface protein. PMID- 12552018 TI - Identification of a TAP-independent, immunoproteasome-dependent CD8+ T-cell epitope in Epstein-Barr virus latent membrane protein 2. AB - We have identified an HLA-A2-restricted CD8(+) T-cell epitope, FLYALALLL, in the Epstein-Barr virus (EBV) latent membrane protein 2 (LMP2), an important target antigen in the context of EBV-associated malignancies. This epitope is TAP independent, like other hydrophobic LMP2-derived epitopes, but uniquely is dependent upon the immunoproteasome for its generation. PMID- 12552019 TI - Human immunodeficiency virus type 1 attachment, coreceptor, and fusion inhibitors are active against both direct and trans infection of primary cells. AB - Inhibitors of human immunodeficiency virus type 1 attachment (CD4-immunoglobulin G subclass 2), CCR5 usage (PRO 140), and fusion (T-20) were tested on diverse primary cell types that represent the major targets both for infection in vivo and for the inhibition of trans infection of target cells by virus bound to dendritic cells. Although minor cell-type-dependent differences in potency were observed, each inhibitor was active on each cell type and trans infection was similarly vulnerable to inhibition at each stage of the fusion cascade. PMID- 12552020 TI - Characterization of tissue tropism determinants of adeno-associated virus type 1. AB - Muscle is an attractive target for gene delivery because of its mass and because vectors can be delivered in a noninvasive fashion. Adeno-associated virus (AAV) has been shown to be effective for muscle-targeted gene transfer. Recent progress in characterization of AAV serotype 1 (AAV1) and AAV6 demonstrated that these two AAV serotypes are far more efficient in transducing muscle than is the traditionally used AAV2. Since all cis elements are identical in these vectors, the potential determinants for their differences in transducing muscle appear to be located within the AAV capsid proteins. In the present study, a series of AAV capsid mutants were generated to identify the major regions affecting AAV transduction efficiency in muscle. Replacement of amino acids 350 to 736 of AAV2 VP1 with the corresponding amino acids from VP1 of AAV1 resulted in a hybrid vector that behaved very similarly to AAV1 in vitro and in vivo in muscle. Characterization of additional mutants carrying smaller regions of the AAV1 VP1 amino acid sequence in the AAV2 capsid protein suggested that amino acids 350 to 430 of VP1 function as a major tissue tropism determinant. Further analysis showed that the heparin binding domain and the major antigenic determinants in the AAV capsid region were not necessary for the efficiency of AAV1 transduction of muscle. PMID- 12552021 TI - Kinetics of virus-specific CD8+ -T-cell expansion and trafficking following central nervous system infection. AB - CD8+ T cells control acute infection of the central nervous system (CNS) by neurotropic mouse hepatitis virus but do not suffice to achieve sterile immunity. To determine the lag between T-cell priming and optimal activity within the CNS, the accumulation of virus-specific CD8+ T cells in the CNS relative to that in peripheral lymphoid organs was assessed by using gamma interferon-specific ELISPOT assays and class I tetramer staining. Virus-specific CD8+ T cells were first detected in the cervical lymph nodes. Expansion in the spleen was delayed and less pronounced but also preceded accumulation in the CNS. The data further suggest peripheral acquisition of cytolytic function, thus enhancing CD8+ -T-cell effector function upon cognate antigen recognition in the CNS. PMID- 12552022 TI - The latency-associated nuclear antigen of Kaposi's sarcoma-associated herpesvirus permits replication of terminal repeat-containing plasmids. AB - The latency-associated nuclear antigen (LANA) of Kaposi's sarcoma-associated herpesvirus can associate with mitotic chromosomes and promote latent episome maintenance and segregation. Here we report that LANA also mediates the replication of plasmid DNAs bearing viral terminal repeats. The predicted secondary structure of LANA's C terminus reveals striking similarity to the known structure of the DNA-binding domain of Epstein-Barr virus EBNA1, despite the absence of primary sequence homology between these proteins, suggesting conservation of the key mechanistic features of latent gammaherpesvirus DNA replication. PMID- 12552023 TI - Molecular chaperone GRP78/BiP interacts with the large surface protein of hepatitis B virus in vitro and in vivo. AB - The proper folding and assembly of viral envelope proteins are mediated by host chaperones. In this study, we demonstrated that an endoplasmic reticulum luminal chaperone GRP78/BiP bound specifically to the pre-S1 domain of the L protein in vitro and in vivo where complete viral particles were secreted, suggesting that GRP78/BiP plays an essential role in the proper folding of the L protein and/or assembly of viral envelope proteins. PMID- 12552025 TI - Success or failure with antiepileptic drug therapy: Beyond empiricism? PMID- 12552024 TI - Molecular characterization of bovine enteric caliciviruses: a distinct third genogroup of noroviruses (Norwalk-like viruses) unlikely to be of risk to humans. AB - Bovine enteric caliciviruses (BoCVs) have been classified in the Norovirus (Norwalk-like virus) genus of the Caliciviridae, raising questions about zoonotic transmission and an animal reservoir for the human Norwalk-like viruses (NLVs), an important cause of nonbacterial gastroenteritis in humans. We examined the genetic relationship of human NLVs to BoCVs that were identified by using reverse transcription-PCR with primer pairs originally designed to detect human NLVs. Polymerase, capsid, and open reading frame 3 (ORF3) gene sequence analyses of BoCVs that were identified from 1976 to 2000 from throughout the United Kingdom showed that BoCVs formed a distinct third genogroup of closely related viruses distinct from the human genogroup I and II NLVs. Evidence was not obtained to support the concept that BoCVs are circulating in humans and pose a threat to human health. PMID- 12552026 TI - Another double trouble. Silent carriers of doublecortin mutations. PMID- 12552027 TI - Practice parameter: treatment of the child with a first unprovoked seizure: Report of the Quality Standards Subcommittee of the American Academy of Neurology and the Practice Committee of the Child Neurology Society. AB - The Quality Standards Subcommittee of the American Academy of Neurology and the Practice Committee of the Child Neurology Society develop practice parameters as strategies for patient management based on analysis of evidence regarding risks and benefits. This parameter reviews published literature relevant to the decision to begin treatment after a child or adolescent experiences a first unprovoked seizure and presents evidence-based practice recommendations. Reasons why treatment may be considered are discussed. Evidence is reviewed concerning risk of recurrence as well as effect of treatment on prevention of recurrence and development of chronic epilepsy. Studies of side effects of anticonvulsants commonly used to treat seizures in children are also reviewed. Relevant articles are classified according to the Quality Standards Subcommittee classification scheme. Treatment after a first unprovoked seizure appears to decrease the risk of a second seizure, but there are few data from studies involving only children. There appears to be no benefit of treatment with regard to the prognosis for long term seizure remission. Antiepileptic drugs (AED) carry risks of side effects that are particularly important in children. The decision as to whether or not to treat children and adolescents who have experienced a first unprovoked seizure must be based on a risk-benefit assessment that weighs the risk of having another seizure against the risk of chronic AED therapy. The decision should be individualized and take into account both medical issues and patient and family preference. PMID- 12552028 TI - How long does it take for partial epilepsy to become intractable? AB - BACKGROUND: Much remains unknown about the natural history of intractable localization-related epilepsy, including how long it typically takes before intractability becomes evident. This information could guide the design of future studies, resolve certain discrepancies in the literature, and provide more accurate information about long-term prognosis. METHODS: Individuals evaluated for resective surgery for refractory localization-related epilepsy were prospectively identified at the time of initial surgical evaluation at seven surgical centers (between 1996 and 2001). The latency time between onset of epilepsy and failure of second medication and history of remission (>/=1 year seizure-free) before surgical evaluation were examined with respect to age at onset, hippocampal atrophy, febrile seizures, and surgical site. RESULTS: In the 333 patients included in the analysis, latency time was 9.1 years (range 0 to 48) and 26% reported a prior remission before surgery. A prior remission of >/=5 years was reported by 8.5% of study participants. Younger age at onset was strongly associated with longer latency time (p < 0.0001) and higher probability of past remission (p < 0.0001). In multivariable analyses, age at onset remained as the most important explanatory variable of both latency time and prior remission. CONCLUSIONS: A substantial proportion of localization-related epilepsy may not become clearly intractable for many years after onset. This is especially true of epilepsy of childhood and early adolescent onset. If prospective studies confirm these findings and the underlying mechanisms behind these associations become understood, this raises the possibility of considering interventions that might interrupt such a process and some day prevent some forms of epilepsy from becoming intractable. PMID- 12552029 TI - The pathological basis of temporal lobe epilepsy in childhood. AB - OBJECTIVE: To characterize the pathologic findings of temporal lobe epilepsy (TLE) in children undergoing temporal lobectomy for refractory seizures and to correlate these findings with clinical presentation. METHODS: The authors reviewed the charts of all children who underwent anterior temporal lobectomy for refractory TLE from 1979 through 1999. A new neuropathologic analysis was performed blinded to clinical features and outcome. RESULTS: Twenty-two children met inclusion criteria. Mean age at onset of epilepsy was 3 years, 7 months (range 1 month to 10 years). Mean age at surgery was 10 years, 11 months (range 1 to 18 years). All patients had complex partial seizures, 48% with secondary generalization. Most had daily seizures. Auras were reported in 45% of patients. Post-resection follow-up averaged 5 years, 2 months (range 2 to 19 years). Seizure-free status was achieved in 41% of patients, and 14% had residual auras only. The most frequent neuropathologic abnormalities were cortical dysplasia (CD) of the temporal neocortex (14 of 22) and mesial temporal sclerosis (MTS) (12 of the 15 children with available hippocampal tissue). These two findings coexisted in seven children. MTS was associated with extra-hippocampal pathology in 8 of 12 (67%) of the cases. CONCLUSIONS: MTS occurs frequently in association with CD in this population of children. The high incidence of dual pathology could explain the early age of seizure onset and high seizure frequency rate observed. TLE in childhood may constitute a different entity than in adults, from both the clinical and neuropathologic perspectives. PMID- 12552030 TI - A dose-comparison trial of topiramate as monotherapy in recently diagnosed partial epilepsy. AB - OBJECTIVE: To evaluate topiramate as monotherapy in adults and children with recently diagnosed, localization-related epilepsy, comparing two dosages of topiramate in a multicenter, randomized, double-blind study. METHODS: Adults and children (>/=3 years of age) were eligible if the maximum interval since epilepsy diagnosis was 3 years and patients had one to six partial-onset seizures during a 3-month retrospective baseline. At study entry, patients (N = 252) were untreated or receiving one antiepileptic drug for less than 1 month. After randomization to 50 or 500 mg/d topiramate (25 or 200 mg/d if weight /= 3) on the mRS (OR 10.3, 95% CI 2.5 to 42.0), increased instrumental disability on the Lawton Instrumental Activities of Daily Living scale (OR 4.9; 95% CI 1.1 to 22.2), reduced quality of life on the Sickness Impact Profile (OR 4.5; 95% CI 1.1 to 18.0), and increased state anxiety on the Spielberger Anxiety Inventory (OR 4.8; 95% CI 1.1 to 20.4). Epilepsy was not associated with cognitive impairment, depression, or subjective life satisfaction. CONCLUSION: Epilepsy occurred in 7% of patients with SAH, was predicted by subdural hematoma and cerebral infarction, and was associated with poor functional recovery and quality of life. Our findings indicate that focal pathology, rather than diffuse injury from hemorrhage, is the principal cause of epilepsy after SAH. PMID- 12552033 TI - Febrile seizures and mesial temporal sclerosis: No association in a long-term follow-up study. AB - OBJECTIVE: To determine whether febrile seizures cause mesial temporal sclerosis (MTS), the occurrence of MTS was evaluated in an unselected series of patients with febrile seizures. METHODS: Twenty-four patients with a prolonged first febrile seizure, 8 with an unprovoked seizure after the first febrile seizure, and 32 age-, sex-, and handedness-matched control subjects with a single simple febrile seizure without later unprovoked seizures were selected from 329 febrile seizure patients followed up prospectively. The occurrence of MTS was evaluated after a mean follow-up time of 12.3 years by MR volumetry of amygdala and hippocampal formation and qualitative analysis of mesial temporal structures. RESULTS: None of the patients had MTS. The mean total volumes of the right and left hippocampal formations and amygdala did not differ significantly between any of the three groups. The qualitative analysis revealed no sclerotic changes in the mesial temporal area. The patients with a prolonged initial febrile seizure had a lower mean right-left volume difference in hippocampal formations than the control subjects, but this had no effect on the outcome. CONCLUSION: The occurrence of MTS following even prolonged febrile seizures is an uncommon event, confirming the good clinical outcome of febrile seizures. PMID- 12552034 TI - Multiple sclerosis: Neurofilament light chain antibodies are correlated to cerebral atrophy. AB - OBJECTIVE: To evaluate markers of axonal damage in CSF and serum of patients with different subtypes of MS in relation to measures of disease progression on MRI. METHODS: In 51 patients with MS (21 relapsing-remitting, 20 secondary progressive, 10 primary progressive), levels of heavy and light neurofilaments (NfH and NfL) and antibodies to neurofilaments (anti-NfL and -NfH) as well as the total immunoglobulin G (IgG) were analyzed. MRI analysis included T2 hyperintense, T1 hypointense, and gadolinium enhancing lesions and markers of cerebral atrophy (ventricular and parenchymal fractions). RESULTS: For the total group, correlations were found between the anti-NfL index and the parenchymal fraction (PF) (r = -0.51, p < 0.001), T2 lesion load (r = 0.41, p < 0.05), ventricular fraction (r = 0.37, p < 0.05), and T1 lesion load (r = 0.37, p < 0.05). For the anti-NfH index, a correlation was found with the PF (r = -0.39, p < 0.05). No correlations were found between the IgG index and MRI measures. CONCLUSIONS: Intrathecal production of anti-NfL antibodies may serve as a marker of tissue damage, particularly axonal loss, in MS. PMID- 12552035 TI - The pentapeptide QYNAD does not block voltage-gated sodium channels. AB - BACKGROUND: An endogenous pentapeptide (Gln-Tyr-Asn-Ala-Asp; QYNAD) that is present at elevated levels in human CSF from patients with demyelinating diseases has been reported to block voltage-gated sodium channels at low (10 micro M) concentrations. Objective : Because of the potential importance of sodium channel blocking activity in demyelinating disorders, this study attempted to determine the sensitivity to QYNAD of different sodium channel subtypes, including Na(v)1.6, the major sodium channel at nodes of Ranvier, and Na(v)1.2, which is expressed in axons with abnormal myelin. METHODS: Sodium channel function was assayed using patch-clamp recordings, both in heterologous expression systems and in intact neurons. RESULTS: QYNAD synthesized in 10 different batches by four different facilities failed to block sodium currents, even at concentrations as high as 500 micro M (50-fold higher than the blocking concentration originally reported). QYNAD had no effect on the currents produced by recombinant Na(v)1.2, Na(v)1.4, Na(v)1.6, and Na(v)1.7 sodium channels or on the sodium currents that are produced by native channels in adult hippocampal or dorsal root ganglion neurons. QYNAD did not interfere with conduction in the optic nerve, a myelinated fiber tract that is often affected in MS. CONCLUSIONS: These experiments do not show any sodium channel blocking effect of QYNAD. The conclusion that QYNAD contributes to the pathophysiology of inflammatory neurologic disorders by blocking voltage-gated sodium channels should therefore be viewed with caution. PMID- 12552036 TI - Anti-Tr antibodies as markers of paraneoplastic cerebellar degeneration and Hodgkin's disease. AB - BACKGROUND: Preliminary studies suggested that anti-Tr antibodies identify patients with paraneoplastic cerebellar degeneration (PCD) and Hodgkin disease (HD). OBJECTIVE: To extend the clinical-immunologic analysis to 28 patients with anti-Tr antibodies. METHODS: Anti-Tr antibodies were detected by immunohistochemistry. A competitive inhibition assay was used to ascertain if anti-Tr antibodies of different sera identify common epitopes. Anti-Tr immunoglobulin G (IgG) subclass distribution was determined by immunohistochemistry using monoclonal antibodies against human IgG isotypes. Tr immunoreactivity was analyzed in tumor sections using biotinylated anti-Tr IgG. RESULTS: Median age of the 28 patients was 61 years (range 14 to 75 years) and 22 were male. A cerebellar syndrome was present in 27 patients and a possible limbic encephalitis in one. HD was diagnosed in 25 patients. No tumor was found in three patients; the autopsy of one of them disclosed severe loss of Purkinje cells without inflammatory infiltrates. Anti-Tr antibodies spontaneously disappeared in all patients without tumor and in 10/10 patients after successful HD treatment. Anti-Tr antibodies were absent in the serum but positive in the CSF of two patients. All positive anti-Tr sera inhibited the immunoreactivity of biotinylated anti-Tr IgG. The predominant isotypes of anti-Tr were IgG1 and IgG3. Only 1 out of the 15 HD samples studied presented anti-Tr positivity that was localized in some Reed-Sternberg cells. CONCLUSIONS: This study confirms the strong association between anti-Tr antibodies and PCD associated with HD. Anti-Tr antibodies from different patients recognize similar epitopes. Unlike other antineuronal antibodies, anti-Tr antibodies can be detected in the CSF but not in the serum and may spontaneously disappear during the follow-up, and Tr immunoreactivity is usually lacking in the tumor. PMID- 12552037 TI - Early onset familial Alzheimer's disease: Mutation frequency in 31 families. AB - BACKGROUND: Three causative genes have been identified for autosomal dominant AD. OBJECTIVE: To determine the proportion of patients with early onset AD with a positive family history accounted for by mutations in these genes. METHODS: A mutational analysis of the amyloid precursor protein (APP), presenilin 1 (PSEN1), and presenilin 2 (PSEN2) genes was performed in 31 probands with probable or definite AD from UK families with an age at onset (AAO) <61 years. RESULTS: The mean AAO was 46.9 years (median 45 years; range 33 to 60 years). The majority of patients (23 of 31; 74%) fulfilled recognized criteria for autosomal dominant inheritance. In 17 (55%) probands the authors identified eight novel PSEN1 sequence variants and eight recognized pathogenic mutations. In 4 (13%) probands the authors identified one novel APP sequence variant (H677R) and two recognized mutations. Thus in this series 21 of 31 (68%) probands were associated with a sequence variant in APP or PSEN1. Nine of the 11 (82%) probands with neuropathologically confirmed AD who additionally fulfilled recognized criteria for autosomal dominant inheritance were associated with a sequence variant in APP or PSEN1. The 10 patients in whom the authors were unable to identify a mutation in APP, PSEN1, or PSEN2 were older than the probands with sequence variants (55.4 vs 44.7 years: p = 0.001). CONCLUSIONS: Sequence variants in APP and PSEN1 accounted for the majority of neuropathologically confirmed autosomal dominant early onset AD; no mutations in PSEN2 were detected. There may be a further genetic factor involved in the etiology of autosomal dominant early onset AD. PMID- 12552038 TI - APOE E4 is a determinant for Alzheimer type pathology in progressive supranuclear palsy. AB - OBJECTIVE: To assess demographic and genetic determinants of Alzheimer type pathology in progressive supranuclear palsy (PSP). METHODS: From a total of 173 pathologically proven cases of PSP in the Society for PSP Brain Bank, 143 patients (mean age = 74.4 years, ranging from 42 to 98 years) were suitable for genetic and pathologic study. Senile plaques (SPs) and neurofibrillary tangles (NFTs) were counted in five cortical areas, Braak stage was given, and APOE genotype was determined from DNA isolated from frozen brain tissue. The APOE allele frequency in PSP with varying degrees of concomitant Alzheimer type pathology was compared with pure PSP and autopsy controls. The relationship of APOE epsilon4 to quantitative pathologic measures was also assessed. RESULTS: Most patients with PSP (103 cases) had either minimal or no Alzheimer type pathology (Braak stage III or less), but 14 patients had many SPs (>20 per low power field) and a Braak stage of IV or higher consistent with pathologic criteria for AD, and 26 patients had many diffuse plaques with minimal neurofibrillary degeneration (Braak stage III or less) consistent with pathologic aging. Alzheimer type pathology was more frequent in women and older individuals with PSP, and 19 of the 40 patients with PSP with Alzheimer type pathology carried at least one APOE epsilon4 allele. The epsilon4 allele frequency was significantly higher in PSP with AD or pathologic aging than in pure PSP, and APOE epsilon4 correlated with the average maximal density of both neocortical SPs and NFTs. CONCLUSION: APOE epsilon4 is a risk factor for Alzheimer type pathology in PSP. Alzheimer type pathology is an independent process unrelated to PSP in cases with both types of pathologies. PMID- 12552039 TI - Apolipoprotein E epsilon4 allele, AD pathology, and the clinical expression of Alzheimer's disease. AB - OBJECTIVE: To test the hypothesis that the APOE epsilon4 allele is associated with the clinical manifestations of AD through an association with the pathologic hallmarks of disease. METHODS: Participants were older Catholic nuns, priests, and brothers who agreed to annual neurologic and neuropsychological evaluation for AD and other common neurologic conditions and brain autopsy at the time of death. There were 77 persons without dementia and 51 with probable AD; 38 participants had one or more epsilon4 alleles. RESULTS: In logistic regression analyses, controlling for age, sex, and education, the epsilon4 allele was strongly associated with the likelihood of clinical AD (odds = 3.46, 95% CI = 1.44 to 8.33). However, controlling for the effect of AD pathology, the association of the epsilon allele with clinical AD was reduced by >50% and was no longer significant (odds = 1.58, 95% CI = 0.56 to 4.43). Similarly, in linear regression analyses, controlling for age, sex, and education, the epsilon4 allele was strongly associated with level of cognitive function proximate to death (regression coefficient = -0.477, p = 0.005). However, after controlling for the effect of AD pathology, the association of the epsilon4 allele with level of cognition was reduced by >80% and was no longer significant (regression coefficient = -0.093). Similar results were found in analyses using separate measures of neuritic plaques, diffuse plaques, and neurofibrillary tangles, and in analyses of five different cognitive systems (episodic memory, semantic memory, working memory, perceptual speed, and visuospatial ability). CONCLUSIONS: The APOE epsilon4 allele appears to be associated with the clinical manifestations of AD through an association with the pathologic hallmarks of AD rather than another mechanism. PMID- 12552040 TI - MRI as a biomarker of disease progression in a therapeutic trial of milameline for AD. AB - OBJECTIVE: To assess the feasibility of using MRI measurements as a surrogate endpoint for disease progression in a therapeutic trial for AD. METHODS: A total of 362 patients with probable AD from 38 different centers participated in the MRI portion of a 52-week randomized placebo-controlled trial of milameline, a muscarinic receptor agonist. The therapeutic trial itself was not completed due to projected lack of efficacy on interim analysis; however, the MRI arm of the study was continued. Of the 362 subjects who underwent a baseline MRI study, 192 subjects underwent a second MRI 1 year later. Hippocampal volume and temporal horn volume were measured from the MRI scans. RESULTS: The annualized percent changes in hippocampal volume (-4.9%) and temporal horn volume (16.1%) in the study patients were consistent with data from prior single-site studies. Correlations between the rate of MRI volumetric change and change in behavioral/cognitive measures were greater for the temporal horn than for the hippocampus. Decline over time was more consistently seen with imaging measures, 99% of the time for the hippocampus, than behavioral/cognitive measures (p < 0.001). Greater consistency in MRI than behavioral/clinical measures resulted in markedly lower estimated sample size requirements for clinical trials. The estimated number of subjects per arm required to detect a 50% reduction in the rate of decline over 1 year are: AD Assessment Scale-cognitive subscale 320; Mini Mental Status Examination 241; hippocampal volume 21; temporal horn volume 54. CONCLUSION: The consistency of MRI measurements obtained across sites, and the consistency between the multisite milameline data and that obtained in prior single-site studies, demonstrate the technical feasibility of using structural MRI measures as a surrogate endpoint of disease progression in therapeutic trials. However, validation of imaging as a biomarker of therapeutic efficacy in AD awaits a positive trial. PMID- 12552041 TI - Telephone screening for amnestic mild cognitive impairment. AB - OBJECTIVES: To evaluate the utility of telephone screening for identifying subjects with amnestic mild cognitive impairment (aMCI) for enrollment in a clinical trial and to identify which elements of the modified Telephone Interview for Cognitive Status (TICS-m) best predicted the in-clinic determination of aMCI. METHODS: Subjects aged >/=65 years with memory complaints responded to an advertisement for a clinical trial by calling a central telephone recruiting agency. To determine eligibility, subjects went through a stepwise selection procedure involving a review of major protocol inclusion and exclusion criteria, followed by administration of the Category Fluency Test (CFT) and then the TICS m. Subjects meeting entry criteria, who obtained a score of /=24 on the Mini-Mental State Examination and a score of 60 years (odds ratio 1.9; 95% CI 1.2 to 2.9; p = 0.003), history of multiple TIAs (odds ratio 2.9; 2.1 to 4.0; p < 0.001), duration of spell 10 minutes, diabetes, weakness, and speech impairment. Among the 30 patients with isolated sensory symptoms lasting /= 14). Twelve of the original 28 items in the SIS version 3.0 composite physical domain were eliminated to produce the SIS-16, with a minimal loss of reliability. As compared to the BI, the SIS-16 contains more difficult items that can differentiate patients with less severe limitations, and therefore has less pronounced ceiling effects. SIS-16 scores were significantly different across Rankin levels 0 to 1, 2, 3, 4, and 5, whereas BI was significantly different only across Rankin levels 0 to 2, 3, 4, and 5. CONCLUSION: Compared to the BI, the SIS-16 is an excellent collection of items suitable for assessing a wide range of physical function limitations of patients with stroke at 1 to 3 months poststroke. Because of a less pronounced ceiling effect, the SIS-16 can differentiate lower levels of disability as compared to the BI. PMID- 12552048 TI - Role of C-nociceptive fibers in the nerve axon reflex-related vasodilation in diabetes. AB - OBJECTIVE: To evaluate the role of the C-nociceptive nerve fibers in nerve axon reflex-related vasodilation. METHODS: Skin vascular reactivity, in response to iontophoresis of acetylcholine and sodium nitroprusside, was evaluated at both the forearm and the foot levels in 13 diabetic neuropathic (DN),11 nonneuropathic (D), and 9 healthy control (C) subjects. The total and nerve axon reflex-related vasodilation were measured by two single-point laser probes. A topical anesthetic was applied on the contralateral forearm and foot, and all the measurements were repeated. RESULTS: Dermal anesthesia resulted in a reduction of the nerve axon reflex-related vasodilation at the forearm in all three groups (C 70.7 +/- 12%, D 59.7 +/- 7%, and DN 73.5 +/- 14%; percentage of reduction over preanesthesia response, mean +/- SEM; p < 0.001) and at the foot in the two nonneuropathic groups (C 74 +/- 10% and D 68.9 +/- 9%; p < 0.001 versus before anesthesia). This reduction was absent at the foot of the neuropathic patients (DN -4 +/- 21%; p = NS versus before anesthesia). A correlation was found between the nerve axon reflex-related response and measurements of nerve function (neuropathy disability score, r = -0.425, p < 0.017; vibration perception threshold, r = -0.527, p < 0.002; Semmes-Weinstein monofilament perception, r = -0.619, p < 0.001). CONCLUSION: The nerve axon reflex-related vasodilation is directly related to the function of the C-nociceptive fibers and is significantly associated with other nerve function measurements. As this is an objective measurement, it has the potential to be used as an alternative to currently employed techniques to evaluate small-fiber function. PMID- 12552049 TI - Sera from Guillain-Barre patients enhance leakage in blood-nerve barrier model. AB - BACKGROUND: In Guillain-Barre syndrome (GBS), the destruction or malfunction of blood-nerve barrier (BNB) has been considered to be the beginning of the disease process. It is unclear whether sera from patients with GBS can open the BNB, and which component of patient sera is most important in the dysregulation of the BNB. METHODS: The authors evaluated the effect of sera from patients with GBS on permeability of an in vitro BNB model using bovine endoneurial microvascular endothelial cells (PnMEC) cultured on the luminal side of a collagen-coated culture insert (pore size: 0.4 micro m). RESULTS: PnMEC monolayers challenged by GBS sera showed significantly lower transendothelial electrical resistance and higher clearance of [carboxyl-(14)C]-inulin with or without complement. Sera with anti-GM1 antibody showed greater loosening of the barrier than others. This effect decreased significantly after incubation with pure GM1 antigen, suggesting the importance of anti-GM1 antibody in BNB dysregulation. Serial analyses of [carboxyl-(14)C]-inulin clearance in four patients disclosed a favorable effect of plasmapheresis in restoring BNB function in some cases. CONCLUSIONS: The authors found an unfavorable effect of sera from patients with GBS on BNB function, supporting involvement of humoral factors causing BNB derangement in the acute stage. Serial evaluation of permeability change using the authors' in vitro system might be useful for the clinical assessment of BNB derangement in individual patients. PMID- 12552050 TI - Oxidative stress in HIV demented patients and protection ex vivo with novel antioxidants. AB - OBJECTIVE: To determine the role of oxidative stress in mediating HIV dementia and to identify novel therapeutic compounds that may block this oxidative stress. METHODS: Brain tissue from patients with HIV encephalitis and macaques with simian immune deficiency virus encephalitis was immunostained for lipid peroxidation. Oxidized proteins in CSF of patients with various stages of HIV dementia were quantitated and we determined whether CSF from these patients could alter mitochondrial function. Several novel compounds with antioxidant effects were screened to determine their relative efficacy in protecting against CSF induced neurotoxicity. RESULTS: Evidence for oxidative stress was present both in brain and in CSF. The presence of oxidized proteins in the CSF and CSF-induced progressive decrease in mitochondrial activity correlated with the severity of cognitive impairment, but only the group of patients with moderate to severe dementia reached statistical significance. L-deprenyl, didox, imidate, diosgenin, and ebselen blocked the CSF-induced toxicity. No effect of trimidox, ruthenium red, or Quercetin was seen. CONCLUSIONS: Increased oxidative stress is present in brain and CSF of HIV-infected patients. There is also an accumulation of toxic substances in the CSF that are capable of inducing oxidative stress. The authors have identified several novel compounds that are capable of blocking the CSF induced toxicity, the therapeutic potential of which is worthy of further exploration. PMID- 12552052 TI - Parinaud syndrome heralding MS. PMID- 12552051 TI - Acute migraine treatment with droperidol: A randomized, double-blind, placebo controlled trial. AB - BACKGROUND: The treatment of a migraine attack can be difficult when first-line medication is unsuccessful and options for parenteral "rescue" therapy are limited. METHODS: A randomized, double-blind, placebo-controlled, dose-ranging, multicenter study was conducted to assess the efficacy and tolerability of droperidol 0.1 mg, 2.75 mg, 5.5 mg, and 8.25 mg for the acute treatment of moderate to severe migraine headache in adults. RESULTS: A total of 331 patients were enrolled; 305 were treated. Headache response at 2 hours was better (p < 0.002) in the treatment groups receiving droperidol IM at doses of 2.75 mg (87%), 5.5 mg (81%), and 8.25 mg (85%) compared with placebo (57%). The percent of patients achieving a pain-free response at 2 hours after treatment was significantly greater than placebo for the droperidol 2.75-mg, 5.5-mg, and 8.25 mg dose groups. The frequency of headache recurrence (within 24 hours) for patients initially responding by 2 hours was lower in patients treated with droperidol than placebo, but differences failed to reach significance. A significantly greater percentage of patients receiving droperidol 2.75 mg reported the elimination of migraine-associated symptoms (nausea, vomiting, photophobia, and phonophobia) than those who received placebo. Although most adverse events were of mild or moderate intensity, anxiety, akathisia, and somnolence were rated as severe in 30% of patients who experienced those symptoms. Hypotension was uncommon. No patient had QT prolongation. PMID- 12552053 TI - Intercaudate nucleus ratio as a linear measure of brain atrophy in multiple sclerosis. AB - Intercaudate nucleus ratio (ICR) is a linear measure of brain atrophy that does not require software application and is independent of image acquisition techniques. The authors examined the relationship between ICR and disability in 190 patients with MS. The results show that ICR correlates with Expanded Disability Status Scale score (r = 0.67; p = 0.0001) and disease duration (r = 0.32; p < 0.01). Intercaudate ratio appears to be a reliable and reproducible linear measure of brain atrophy and correlates with disability and disease duration in MS. PMID- 12552054 TI - Posterior leukoencephalopathy following intrathecal chemotherapy with MRA documented vasospasm. AB - Posterior leukoencephalopathy syndromes have been reported with hypertension, and immunosuppressive and chemotherapy agents. Cerebral vasospasm on MR angiography (MRA) has been noted in cases due to eclampsia. The authors report a case of Balint syndrome with irreversible posterior leukoencephalopathy on MRI following intrathecal methotrexate and cytarabine. Hypertension was not present. Diffuse, reversible arterial irregularities consistent with vasospasm were present on MRA during the acute illness. PMID- 12552055 TI - Somatic mosaicism and variable penetrance in doublecortin-associated migration disorders. AB - X-linked isolated lissencephaly sequence (XLIS) and subcortical band heterotopia (SBH) are allelic disorders caused by mutations in the doublecortin (DCX) gene. This genetic analysis of seven families revealed four novel mutations in the DCX gene. The authors detected a high rate of somatic mosaicism in male and female patients with variable penetrance of bilateral SBH including nonpenetrance in a heterozygous woman. In addition, the authors implemented prenatal diagnosis in a family with SBH/XLIS. PMID- 12552056 TI - The use of topiramate in refractory status epilepticus. AB - In cases of refractory status epilepticus (RSE) unresponsive to sequential trials of multiple agents, a suspension of topiramate administered via nasogastric tube was effective in aborting RSE, including one patient in a prolonged pentobarbital coma. Effective dosages ranged from 300 to 1,600 mg/d. Except for lethargy, no adverse events were reported. PMID- 12552057 TI - Neonatal dopa-responsive extrapyramidal syndrome in twins with recessive GTPCH deficiency. AB - The authors report two twin sisters, age 15 years, with recessive GTP cyclohydrolase deficiency, who presented with neonatal onset of rigidity, tremor, and dystonia but with no other symptoms suggestive of a diffuse CNS involvement. The plasma phenylalanine levels were normal. Treatment with L-dopa/carbidopa, started at age 1 year, was associated with sustained recovery from all neurologic signs. The patients were homozygous for a new recessive mutation in the GHI gene. PMID- 12552058 TI - Toxic neuropathy in patients with pre-existing neuropathy. AB - The authors report significant worsening of a pre-existing neuropathy in six patients who received "non-toxic" dosages of known neurotoxic agents. Before treatment, baseline total neuropathy score (TNS) averaged 9.5 (range 0 to 19). After chemotherapy (Taxol [125 to 175 mg/m(2) x 4]; vincristine [2 to 5 mg]; cisplatin [40 mg/m(2) x 8]; and thalidomide [60 g]), the TNS averaged 22 (range 13 to 29). The authors conclude that functionally disabling toxic neuropathy can occur in patients with pre-existing neuropathy at standard doses. PMID- 12552059 TI - Paramyotonia congenita with an SCN4A mutation affecting cardiac repolarization. AB - Paramyotonia congenita (PC) is linked to mutations of the skeletal muscle voltage gated sodium channel alpha-subunit gene SCN4A. The authors report a family where the proband and three of her four children have PC (mutation R1448C) and present repolarization abnormalities at electrocardiogram. They demonstrate that the SCN4A alpha-subunit gene is expressed in normal human heart. Cardiac consequences of mutations of the SCN4A gene may be insignificant in standard conditions, but critical if patients with PC are treated with drugs inducing QT prolongation. PMID- 12552060 TI - Is hypertension associated with hemifacial spasm? PMID- 12552061 TI - Lithium-induced periodic alternating nystagmus. PMID- 12552062 TI - Pneumocephalus by Valsalva's maneuver. PMID- 12552063 TI - Transient ophthalmoplegia following envenomation by the scorpion Mesobuthus eupeus. PMID- 12552064 TI - Sonographic demonstration of temporal arteritis. PMID- 12552065 TI - Multiphasic demyelinating disease involving central and peripheral nervous system in a child. PMID- 12552066 TI - Spinal meningioma causing diffuse leptomeningeal enhancement. PMID- 12552068 TI - Tau and Abeta42 protein in CSF of patients with frontotemporal degeneration. PMID- 12552069 TI - Symptoms of B(12) deficiency can occur in women of childbearing age supplemented with folate. PMID- 12552070 TI - Is it time to replace the Wada test? PMID- 12552071 TI - Seizures in children with fever: Generally good outcome. PMID- 12552073 TI - Effects of patents and licenses on the provision of clinical genetic testing services. AB - The growth of patents that include genetic sequences has been accompanied by concern about their impact on the ability of physicians to provide clinical genetic testing services and to perform research. Therefore, we conducted a survey of clinical laboratory directors that perform DNA-based genetic tests to examine potential effects. We performed a telephone survey between July and September in 2001 of all laboratory directors in the United States who were members of the Association for Molecular Pathology or who were listed on the GENETESTS:org website. One hundred thirty-two of 211 (63%) laboratory directors were interviewed. Ten of these were excluded because they did not conduct DNA based genetic tests. Almost all performed genetic tests for clinical purposes. Half performed tests for research purposes as well. Twenty-five percent of respondents reported that they had stopped performing a clinical genetic test because of a patent or license. Fifty-three percent of respondents reported deciding not to develop a new clinical genetic test because of a patent or license. In total, respondents were prevented from performing 12 genetic tests, and all of these tests were among those performed by a large number of laboratories. We found 22 patents that were relevant to the performance of these 12 tests. Fifteen of the 22 patents (68%) are held by universities or research institutes, and 13 of the 22 patents (59%) were based on research funded by the United States Government. Overall, respondents reported that their perceptions of the effects of patents on the cost, access, and development of genetic tests, or data sharing among researchers, were negative. In contrast, most respondents felt that patents did not have an effect on the quality of testing. We conclude that patents and licenses have had a significant effect on the ability of clinical laboratories to develop and provide genetic tests. Furthermore, our findings suggest that clinical geneticists feel that their research is inhibited by patents. The effects of patents and licenses on patients' access to tests, and the costs and quality thereof, remains to be determined. PMID- 12552074 TI - Accurate and reproducible gene expression profiles from laser capture microdissection, transcript amplification, and high density oligonucleotide microarray analysis. AB - Gene expression profiling using high density oligonucleotide arrays is a powerful method to generate an unbiased survey of a cell's transcriptional landscape. Increasingly complex biological questions require that this approach be applicable to the small numbers of cells that are obtained from sources such as laser capture microdissection (LCM) of solid tissues. In this report, we demonstrate that two rounds of transcript amplification can generate accurate and reproducible gene expression profiles using high density oligonucleotide microarrays, starting with as little as 10 ng of total RNA. Biased amplification of the 3' end of transcripts does not have a major impact on the overall transcript profile due to the 3' bias of probe sets incorporated in the array design. Furthermore, greater than 95% of all genes detected demonstrate less than a twofold difference in expression when independent tissue dissections of identical cell populations are compared. The accuracy and technical reproducibility of the method suggests that expression profiling using transcript amplification and high density oligonucleotide microarrays can be used on a routine basis. PMID- 12552075 TI - Development of real-time PCR assays for the quantitative detection of Epstein Barr virus and cytomegalovirus, comparison of TaqMan probes, and molecular beacons. AB - Human Epstein-Barr virus (EBV) and cytomegalovirus (CMV) can cause serious complications in immunocompromised patients. Rapid diagnosis of EBV and CMV infection is critical in the management of the disease so that anti-viral therapy can be started early. Here we describe the development of real-time PCR assays using TaqMan probes and molecular beacons and compare the performance of both assays with a well-established, validated, gel-based PCR method for the quantification of EBV and CMV in patients' samples. The TaqMan and molecular beacon assays were linear between 10 to 10(7) viral genomes/reaction. Both assays generated calibration curves with strong correlation and low intra-assay and interassay variation. Results of EBV and CMV viral load determination inpatient samples obtained by the gel-based and real-time PCR were very similar. The real time PCR assays showed increases in viral load before clinical measures of viral disease and decreases in viral load during anti-viral therapy in two of six pediatric patients. The data indicate that these TaqMan and molecular beacon approaches are accurate, rapid, and reliable assays for the diagnosis and monitoring of EBV and CMV infections in patients. PMID- 12552076 TI - Method for optimizing pulsed-field gel electrophoresis banding pattern data. AB - The genomic DNA of 47 strains of TSST-1 toxin-producing Staphylococcus aureus were cleaved with SmaI restriction endonuclease and resolved in an agarose gel by pulsed-field gel electrophoresis (PFGE). An algorithm was designed to standardize the band weights or brightness (trace quantity) produced to a bounded region between 0 and 1 regardless of DNA fragment size while simultaneously reducing gel to-gel variability. The algorithm allows for classification of isolates by band intensity as well as DNA mobility without a numerical hierarchy of band intensity that is caused by ranging DNA fragment lengths. On analysis many isolates were classified as separate entities on the basis of DNA co-migration only. Isolates differing by only DNA co-migration were subjected to a second digestion with restriction enzyme SacII. These isolates were characterized similarly to the standardized trace quantity analysis of SmaI PFGE patterns. The standardization method proposed in this article permits characterization of isolates on the basis of band differences, regardless of DNA co-migration, thus increasing the discriminatory power (0.79 to 0.89) of PFGE by increasing band-associated information. An established unbiased approach to the partitioning of data were also explored. PMID- 12552077 TI - Development of a universal probe for electronic microarray and its application in characterization of the Staphylococcus aureus polC gene. AB - Electronic microarray technology is an exceptionally accurate and effective technique for detecting and defining single nucleotide polymorphisms (SNPs) in DNA sequences. Target oligonucleotides are electronically addressed to a gel matrix containing streptavidin to which biotinylated polymerase chain reaction (PCR) amplicons are bound. Typically, a fluorescent-labeled reporter oligonucleotide specific for each locus of interest is hybridized and reported. We detail the development of a universal reporter system to replace the standard method that is used to detect many different sequences accurately. The universal reporter eliminates the need to synthesize specific labeled reporters for each SNP sequence thereby dramatically reducing the cost and time required for assay development. The feasibility of this approach was demonstrated by successfully analyzing eight SNPs distributed within a highly variable 1-kb region of the polC gene from six isolates of Staphylococcus aureus. PMID- 12552078 TI - Towards quantitative mRNA analysis in paraffin-embedded tissues using real-time reverse transcriptase-polymerase chain reaction: a methodological study on lymph nodes from melanoma patients. AB - Improved extraction techniques combined with sensitive real-time reverse transcriptase-polymerase chain reaction may allow detection of mRNA in formalin fixed, paraffin-embedded (FFPE) materials, but the factors affecting mRNA quantification in clinical material using these methods have not been systematically analyzed. We designed analyses using real-time reverse transcriptase-polymerase chain reaction for quantification of MART-1, beta-actin, and beta(2)-microglobulin mRNAs. The analytical intra- and interassay imprecision (coefficient of variation) was in the range 10 to 20% for all three genes studied. Using these protocols, we studied the influence of tissue autolysis and length of formalin-fixation on mRNA detection in metastatic melanoma. Delay in freezing reduced detectable mRNA, although this was less than predicted and mostly occurred early in autolysis. MART-1, beta-actin, and beta(2)-microglobulin mRNAs were consistently detected in FFPE metastatic melanoma even after fixation for up to 3 weeks, although the total mRNA detected was markedly reduced in fixed compared with fresh tissues (up to 99%). Quantification of MART-1 was, however, possible if this was expressed relative to a housekeeping gene. The polymerase chain reaction product from FFPE tissues could be increased up to 100-fold amplifying short (<136 bp) compared with long amplicons. Variations in time before tissue processing and in fixation length seem to be less important sources of imprecision than previously assumed. Our findings suggest that quantitative analysis of mRNA in archive and routine diagnostic tissues may be possible. PMID- 12552079 TI - Analysis of common mutations in the galactose-1-phosphate uridyl transferase gene: new assays to increase the sensitivity and specificity of newborn screening for galactosemia. AB - Classical galactosemia is a genetic disease caused by mutations in the galactose 1-phosphate uridyl transferase (GALT) gene. Prospective newborn screening for galactosemia is routine and utilizes the universally collected newborn dried blood specimen on filter paper. Screening for galactosemia is achieved through analysis of total galactose (galactose and galactose-1-phosphate) and/or determining the activity of the GALT enzyme. While this approach is effective, environmental factors and the high frequency of the Duarte D2 mutation (N314D) does lead to false positive results. Using DNA derived from the original newborn dried blood specimen and Light Cycler technology a panel of five assays able to detect the four most frequently encountered classical galactosemia alleles (Q188R, S135L, K285N, and L195P) and the N314D Duarte variant mutation are described. The five assays are performed simultaneously using common conditions. Including DNA preparation, set-up, amplification, and analysis the genotype data for all five loci is obtained in less than 2 hours. The assays are easily interpreted and amenable to high-throughput newborn screening. Mutational analysis is useful to reduce false positive results, differentiate D/G mixed heterozygotes from classical galactosemia, and to clearly identify a very high percentage of those affected by classical galactosemia. PMID- 12552080 TI - Amplification and overexpression of the EMS 1 oncogene, a possible prognostic marker, in human hepatocellular carcinoma. AB - DNA amplification in cancer cells frequently involves oncogenes whose increased expression confers a selective advantage on tumor cell growth. In an attempt to identify novel oncogenes involved in hepatocarcinogenesis, representational difference analysis (RDA) was performed using DNA from a primary human hepatocellular carcinoma (HCC) that showed high-level DNA amplifications on chromosomes 1p32 and 11q13 by comparative genomic hybridization. Ten amplification fragments were isolated by RDA, and when used to probe Southern blots of tumor DNA, there was a 5- to 50-fold increase in hybridization intensity relative to normal DNA. The sequence of one amplification product matched that of the EMS1 oncogene, which is located on chromosome 11q13 and is amplified in other cancers. We detected EMS1 amplification in 3 of 17 primary HCC. Overexpression of EMS1 mRNA was observed in 12 of 14 HCC cell lines in the absence of gene amplification or an increased copy-number of the gene. The EMS1 gene encodes cortactin, a cortical actin-associated protein that is a substrate for Src kinase and is involved in cytoskeleton organization. Alterations of the EMS1 gene that lead to overexpression of cortactin may be associated with tumor development in HCC. EMS1 amplification and overexpresion is indicative of unfavorable prognosis in several cancers and may have similar prognostic implications in liver cancer. PMID- 12552081 TI - Quantitative intra-individual monitoring of BCR-ABL transcript levels in archival bone marrow trephines of patients with chronic myeloid leukemia. AB - We established a quantitative real-time RT-PCR assay for the detection of chimeric BCR-ABL transcripts in archival formalin-fixed bone marrow trephines, both acrylate-embedded and paraffin-embedded. This new methodology enables determination of transcript levels in direct comparison to histopathological findings and therapeutic interventions during the time course of the disease in a retrospective and a prospective manner. We found an excellent correlation between the quantitative molecular data and the morphological evaluation as well as the clinical outcome for a cohort of chronic myeloid leukemia patients (n = 10). To the best of our knowledge, this is the first study demonstrating the feasibility of large-scale quantitative expression analysis in archival bone marrow trephines for monitoring molecular markers over several years or even decades. PMID- 12552084 TI - The missing link between thermodynamics and structure in F1-ATPase. AB - F(1)F(o)-ATP synthase is the enzyme responsible for most of the ATP synthesis in living systems. The catalytic domain F(1) of the F(1)F(o) complex, F(1)-ATPase, has the ability to hydrolyze ATP. A fundamental problem in the development of a detailed mechanism for this enzyme is that it has not been possible to determine experimentally the relation between the ligand binding affinities measured in solution and the different conformations of the catalytic beta subunits (beta(TP), beta(DP), beta(E)) observed in the crystal structures of the mitochondrial enzyme, MF(1). Using free energy difference simulations for the hydrolysis reaction ATP+H(2)O --> ADP+P(i) in the beta(TP) and beta(DP) sites and unisite hydrolysis data, we are able to identify beta(TP) as the "tight" (K(D) = 10(-12) M, MF(1)) binding site for ATP and beta(DP) as the "loose" site. An energy decomposition analysis demonstrates how certain residues, some of which have been shown to be important in catalysis, modulate the free energy of the hydrolysis reaction in the beta(TP) and beta(DP) sites, even though their structures are very similar. Combined with the recently published simulations of the rotation cycle of F(1)-ATPase, the present results make possible a consistent description of the binding change mechanism of F(1)-ATPase at an atomic level of detail. PMID- 12552083 TI - Small ubiquitin-related modifier-1 modification mediates resolution of CREB dependent responses to hypoxia. AB - Phosphorylation-dependent ubiquitination combined with proteasomal degradation of transcriptional regulators is a recently appreciated mechanism for control of a number of inflammatory genes. Far less is known about the counterregulatory mechanisms that repress transcriptional activity in these pathways during resolution. Here, we investigated the transient nature of hypoxia-induced tumor necrosis factor (TNF)alpha in T84 cells, a process we have previously shown to involve phosphorylation-dependent degradation of the cAMP-response element binding protein (CREB). Initial studies indicate hypoxia-induced TNFalpha to be a transient event, the resolution of which is associated with the appearance of a higher molecular weight modified form of CREB. Gene array analysis of mRNA derived from hypoxic cells identified a time-dependent induction of small ubiquitin-related modifier (SUMO)-1 mRNA. In prolonged hypoxia, CREB is posttranslationally modified by SUMO-1. Furthermore, SUMO-1 overexpression stabilizes CREB in hypoxia and enhances CREB-dependent reporter gene activity. Site-directed mutagenesis of lysine residues K285 and K304 identifies them as SUMO acceptors in vivo and in vitro. Mutation of K304 also results in loss of CREB nuclear localization, implying a role for SUMO-1 modification at this site in the subcellular localization of CREB. Thus, in prolonged hypoxia, CREB is modified by association with SUMO-1. Furthermore, we hypothesize that such an event stabilizes and promotes nuclear localization of CREB and thus complements an endogenous resolution phase for hypoxia-induced inflammatory processes. PMID- 12552085 TI - Uptake and antifungal activity of oligonucleotides in Candida albicans. AB - Candida albicans is a significant cause of disease in immunocompromised humans. Because the number of people infected by fungal pathogens is increasing, strategies are being developed to target RNAs in fungi. This work shows that oligonucleotides can serve as therapeutics against C. albicans. In particular, oligonucleotides are taken up from cell culture medium in an energy-dependent process. After uptake, oligonucleotides, including RNA, remain mostly intact after 12 h in culture. For culture conditions designed for mammalian cells, intracellular concentrations of oligonucleotides in C. albicans exceed those in COS-7 mammalian cells, suggesting that uptake can provide selective targeting of fungi over human cells. A 19-mer 2'OMe (oligonucleotide with a 2'-O-methyl backbone) hairpin is described that inhibits growth of a C. albicans strain at pH < 4.0. This pH is easily tolerated in some parts of the body subject to C. albicans infections. In vivo dimethyl sulfate modification of ribosomal RNA and the decreased rate of protein synthesis suggest that this hairpin's activity may be due to targeting the ribosome in a way that does not depend on base pairing. Addition of anti-C. albicans oligonucleotides to COS-7 mammalian cells has no effect on cell growth. Evidently, oligonucleotides can selectively serve as therapeutics toward C. albicans and, presumably, other pathogens. Information from genome sequencing and functional genomics studies on C. albicans and other pathogens should allow rapid design and testing of other approaches for oligonucleotide therapies. PMID- 12552086 TI - Determination of the transcriptome of Vibrio cholerae during intraintestinal growth and midexponential phase in vitro. AB - Vibrio cholerae is the etiologic bacterial agent of cholera, a severe diarrheal disease endemic in much of the developing world. The V. cholerae genome contains 3,890 genes distributed between a large and a small chromosome. Although the large chromosome encodes the majority of recognizable gene products and virulence determinants, the small chromosome carries a disproportionate number of hypothetical genes. Thus, little is known about the role of the small chromosome in the biology of this organism or other Vibrio species. We have used the rabbit ileal loop model of V. cholerae infection to obtain in vivo-grown cells under near midexponential conditions in the small-intestinal environment. We compared the global transcriptional pattern of these in vivo-grown cells to those grown to midexponential phase in rich medium under aerobic conditions. Under both conditions, the genes showing the highest levels of expression reside primarily on the large chromosome. However, a shift occurs in vivo that results in many more small chromosomal genes being expressed during growth in the intestine. Our analysis further suggests that nutrient limitation (particularly iron) and anaerobiosis are major stresses experienced by V. cholerae during growth in the rabbit upper intestine. Finally, relative to in vitro growth, the intestinal environment significantly enhanced expression of several virulence genes, including those involved in phenotypes such as motility, chemotaxis, intestinal colonization, and toxin production. PMID- 12552087 TI - HLA-G gene repression is reversed by demethylation. AB - The HLA-G molecule plays an important role in immune tolerance, protecting the fetus from maternal immune attack, and probably contributes to graft tolerance and tumor escape from the host immune system. HLA-G expression is tightly regulated and involves mechanisms acting in part at the transcriptional level. Nevertheless, almost all regulatory sequences that govern constitutive and inducible HLA class I gene transcription are disrupted in the HLA-G gene promoter, suggesting an unusual regulatory process. In further investigating the molecular mechanisms of HLA-G gene activation, we evaluated the influence of epigenetic mechanisms on seven HLA-G-negative cell lines that exhibit various phenotypes. Exposure of cells to histone deacetylase inhibitors, or to the demethylating agent 5-aza-2'-deoxycytidine, revealed that HLA-G gene transcription is inhibited by DNA methylation. Reversal of methylation-mediated repression may directly induce HLA-G cell-surface expression, supporting the idea that HLA-G might be activated by such a mechanism during malignancy, inflammation, and allogenic reactions. PMID- 12552088 TI - Comparison of mouse and human genomes followed by experimental verification yields an estimated 1,019 additional genes. AB - A primary motivation for sequencing the mouse genome was to accelerate the discovery of mammalian genes by using sequence conservation between mouse and human to identify coding exons. Achieving this goal proved challenging because of the large proportion of the mouse and human genomes that is apparently conserved but apparently does not code for protein. We developed a two-stage procedure that exploits the mouse and human genome sequences to produce a set of genes with a much higher rate of experimental verification than previously reported prediction methods. RT-PCR amplification and direct sequencing applied to an initial sample of mouse predictions that do not overlap previously known genes verified the regions flanking one intron in 139 predictions, with verification rates reaching 76%. On average, the confirmed predictions show more restricted expression patterns than the mouse orthologs of known human genes, and two-thirds lack homologs in fish genomes, demonstrating the sensitivity of this dual-genome approach to hard-to-find genes. We verified 112 previously unknown homologs of known proteins, including two homeobox proteins relevant to developmental biology, an aquaporin, and a homolog of dystrophin. We estimate that transcription and splicing can be verified for >1,000 gene predictions identified by this method that do not overlap known genes. This is likely to constitute a significant fraction of the previously unknown, multiexon mammalian genes. PMID- 12552089 TI - Direct detection of antibody-antigen binding using an on-chip artificial pore. AB - We demonstrate a rapid and highly sensitive all-electronic technique based on the resistive pulse method of particle sizing with a pore to detect the binding of unlabeled antibodies to the surface of latex colloids. Here, we use an on-chip pore to sense colloids derivatized with streptavidin and measure accurately their diameter increase on specific binding to several different types of antibodies. We show the sensitivity of this technique to the concentration of free antibody and that it can be used to perform immunoassays in both inhibition and sandwich configurations. Overall, our technique does not require labeling of the reactants and is performed rapidly by using very little solution, and the pore itself is fabricated quickly and inexpensively by using soft lithography. Finally, because this method relies only on the volume of bound ligand, it can be generally applied to detecting a wide range of ligand-receptor binding reactions. PMID- 12552090 TI - Reactivity-based one-pot total synthesis of fucose GM1 oligosaccharide: a sialylated antigenic epitope of small-cell lung cancer. AB - The total synthesis of the sialic acid-containing antigenic epitope fucose GM(1) (Fuc-GM(1)) by an improved reactivity-based one-pot synthetic strategy is reported. Based on a thioglycoside reactivity database, three saccharide building blocks, 3, 4, and 5, were designed and prepared to incorporate a descending order of reactivity toward thiophilic activation. Using the reactivity-based one-pot synthetic method, the fully protected Fuc-GM(1) glycoside 2 was furnished in a facile manner, which was globally deprotected to give the Fuc-GM(1) glycoside 1. In addition, using the promoter system 1 (benzensulfinyl)piperidinetrifluoromethanesulfonic anhydride, the product yield was improved and the reaction time was reduced in comparison with the N iodosuccinimidetrifluoromethanesulfonic acid- and dimethyl (thiomethyl) sulfonium trifluoromethanesulfonate-promoted systems. PMID- 12552091 TI - Estrogen inhibits GH signaling by suppressing GH-induced JAK2 phosphorylation, an effect mediated by SOCS-2. AB - Oral estrogen administration attenuates the metabolic action of growth hormone (GH) in humans. To investigate the mechanism involved, we studied the effects of estrogen on GH signaling through Janus kinase (JAK)2 and the signal transducers and activators of transcription (STATs) in HEK293 cells stably expressing the GH receptor (293GHR), HuH7 (hepatoma) and T-47D (breast cancer) cells. 293GHR cells were transiently transfected with an estrogen receptor-alpha expression plasmid and luciferase reporters with binding elements for STAT3 and STAT5 or the beta casein promoter. GH stimulated the reporter activities by four- to sixfold. Cotreatment with 17beta-estradiol (E(2)) resulted in a dose-dependent reduction in the response of all three reporters to GH to a maximum of 49-66% of control at 100 nM (P < 0.05). No reduction was seen when E(2) was added 1-2 h after GH treatment. Similar inhibitory effects were observed in HuH7 and T-47D cells. E(2) suppressed GH-induced JAK2 phosphorylation, an effect attenuated by actinomycin D, suggesting a requirement for gene expression. Next, we investigated the role of the suppressors of cytokine signaling (SOCS) in E(2) inhibition. E(2) increased the mRNA abundance of SOCS-2 but not SOCS-1 and SOCS-3 in HEK293 cells. The inhibitory effect of E(2) was absent in cells lacking SOCS-2 but not in those lacking SOCS-1 and SOCS-3. In conclusion, estrogen inhibits GH signaling, an action mediated by SOCS-2. This paper provides evidence for regulatory interaction between a sex steroid and the GHJAKSTAT pathway, in which SOCS-2 plays a central mechanistic role. PMID- 12552092 TI - Coordinate inhibition of expression of several genes for protein subunits of human nuclear RNase P. AB - The deliberate inhibition of expression of one of the protein subunits (Rpp38) of human nuclear RNase P is achievable by using external guide sequence (EGS) technology. Both the protein product and the mRNA are greatly reduced 24 h after transient transfection with a gene coding for an appropriate EGS. Control experiments indicated that four other protein subunits of RNase P and their RNAs are also inhibited with no external manipulation. The remaining RNase P proteins, their mRNAs, and the RNA subunit of RNase P all are unchanged. Several short nucleotide sequences adjacent to the ORFs for the inhibited genes are similar and could be targets for transcriptional repression. The explanation of coordinate inhibition of the expression of the product of one particular gene by the transfection of an EGS (or RNA interference) requires some care in terms of interpreting phenotypic effects because, in our case, several gene products that are not targeted are also inhibited. PMID- 12552093 TI - Understanding neural system dynamics through task modulation and measurement of functional MRI amplitude, latency, and width. AB - Estimates of hemodynamic amplitude, delay, and width were combined to investigate system dynamics involved in lexical decision making. Subjects performed a lexical decision task using word and nonword stimuli rotated 0 degrees, 60 degrees, or 120 degrees. Averaged hemodynamic responses to repeated stimulation were fit to a Gamma-variate function convolved with a heavyside function of varying onset and duration to estimate each voxel's activation delay and width. Consistent with prolonged reaction times for the rotated stimuli and nonwords, the motor cortex showed delayed hemodynamic onset for both conditions. Language areas such as the lingual gyrus, middle temporal gyrus, fusiform gyrus, and precuneus all showed delayed hemodynamic onsets to rotated stimuli but not to nonword stimuli. The inferior frontal gyrus showed both increased onset latency for rotated stimuli and a wider hemodynamic response to nonwords, consistent with prolonged processing in this area during the lexical decision task. Phonological processing areas such as superior temporal and angular gyrus showed no delay or width difference for rotated stimuli. These results suggest that phonological routes but not semantic routes to the lexicon can proceed regardless of stimulus orientation. This study demonstrates the utility of estimating hemodynamic delay and width in addition to amplitude allowing for more quantitative measures of brain function such as mental chronometry. PMID- 12552094 TI - Flow, NO, and atherogenesis. PMID- 12552095 TI - Hysteresis meets the cell cycle. PMID- 12552096 TI - Gene expression and viral prodution in latently infected, resting CD4+ T cells in viremic versus aviremic HIV-infected individuals. AB - The presence of HIV-1 in latently infected, resting CD4(+) T cells has been clearly demonstrated in infected individuals; however, the extent of viral expression and the underlying mechanisms of the persistence of HIV-1 in this viral reservoir have not been fully delineated. Here, we show that resting CD4(+) T cells from the majority of viremic patients are capable of producing cell-free HIV-1 spontaneously ex vivo. The levels of HIV-1 released by resting CD4(+) T cells were not significantly reduced in the presence of inhibitors of cellular proliferation and viral replication. However, resting CD4(+) T cells from the majority of aviremic patients failed to produce virions, despite levels of HIV-1 proviral DNA and cell-associated HIV-1 RNA comparable to viremic patients. The DNA microarray analysis demonstrated that a number of genes involving transcription regulation, RNA processing and modification, and protein trafficking and vesicle transport were significantly upregulated in resting CD4(+) T cells of viremic patients compared to those of aviremic patients. These results suggest that active viral replication has a significant impact on the physiologic state of resting CD4(+) T cells in infected viremic patients and, in turn, allows release of HIV-1 without exogenous activation stimuli. In addition, given that no quantifiable virions were produced by the latent viral reservoir in the majority of aviremic patients despite the presence of cell-associated HIV-1 RNA, evidence for transcription of HIV-1 RNA in resting CD4(+) T cells of aviremic patients should not necessarily be taken as direct evidence for ongoing viral replication during effective therapy. PMID- 12552097 TI - beta-Arrestin-mediated PDE4 cAMP phosphodiesterase recruitment regulates beta adrenoceptor switching from Gs to Gi. AB - Phosphorylation of the beta(2) adrenoreceptor (beta(2)AR) by cAMP-activated protein kinase A (PKA) switches its predominant coupling from stimulatory guanine nucleotide regulatory protein (G(s)) to inhibitory guanine nucleotide regulatory protein (G(i)). beta-Arrestins recruit the cAMP-degrading PDE4 phosphodiesterases to the beta(2)AR, thus controlling PKA activity at the membrane. Here we investigate a role for PDE4 recruitment in regulating G protein switching by the beta(2)AR. In human embryonic kidney 293 cells overexpressing a recombinant beta(2)AR, stimulation with isoprenaline recruits beta-arrestins 1 and 2 as well as both PDE4D3 and PDE4D5 to the receptor and stimulates receptor phosphorylation by PKA. The PKA phosphorylation status of the beta(2)AR is enhanced markedly when cells are treated with the selective PDE4-inhibitor rolipram or when they are transfected with a catalytically inactive PDE4D mutant (PDE4D5-D556A) that competitively inhibits isoprenaline-stimulated recruitment of native PDE4 to the beta(2)AR. Rolipram and PDE4D5-D556A also enhance beta(2)AR-mediated activation of extracellular signal-regulated kinases ERK12. This is consistent with a switch in coupling of the receptor from G(s) to G(i), because the ERK12 activation is sensitive to both inhibitors of PKA (H89) and G(i) (pertussis toxin). In cardiac myocytes, the beta(2)AR also switches from G(s) to G(i) coupling. Treating primary cardiac myocytes with isoprenaline induces recruitment of PDE4D3 and PDE4D5 to membranes and activates ERK12. Rolipram robustly enhances this activation in a manner sensitive to both pertussis toxin and H89. Adenovirus mediated expression of PDE4D5-D556A also potentiates ERK12 activation. Thus, receptor-stimulated beta-arrestin-mediated recruitment of PDE4 plays a central role in the regulation of G protein switching by the beta(2)AR in a physiological system, the cardiac myocyte. PMID- 12552098 TI - Transcriptional and posttranscriptional control of hepatitis B virus gene expression. AB - Hepatitis B virus (HBV) infects humans and certain nonhuman primates. Viral clearance and acute disease are associated with a strong, polyclonal, multispecific cytotoxic T lymphocyte response. Infiltrating T cells, as well as other activated inflammatory cells, produce cytokines that can regulate hepatocellular gene expression. Using an HBV transgenic mouse model, our laboratory has previously demonstrated that adoptive transfer of HBV-specific cytotoxic T lymphocytes or injection of IL-2 can noncytopathically inhibit HBV gene expression by a posttranscriptional IFN-gamma- and/or tumor necrosis factor alpha-dependent mechanism. Here, we report that HBV gene expression can also be controlled at the posttranscriptional level during persistent lymphocytic choriomeningitis virus infection. In contrast, it is controlled at the transcriptional level during acute murine cytomegalovirus infection or after repetitive polyinosinic-polycytidylic acid injection. Finally, we show that transcriptional inhibition of HBV is associated with changes in liver-specific gene expression. These results elucidate pathways that regulate the viral life cycle and suggest additional approaches for the treatment of chronic HBV infection. PMID- 12552100 TI - Impacts of anthropogenic and natural NO(x) sources over the U.S. on tropospheric chemistry. AB - We evaluate the impact of anthropogenic and natural NO(x) sources over the contiguous United States on tropospheric NO(x) and O(3) levels by using a global 3D chemical transport model. The effects of major U.S. surface NO(x) emission sources (including anthropogenic, biomass burning, and soil emissions) are compared with that of lightning-produced NO(x). Summer lightning is shown to play a dominant role in controlling NO(x) and O(3) concentrations in the middle and upper troposphere, despite the fact that fossil-fuel burning represents the largest source of NO(x) over the U.S. Furthermore, the effect of regional U.S. lightning is propagated through large areas of the Northern Hemisphere by atmospheric circulation. The results reveal that a thorough assessment of atmospheric NO(x) emission sources and their impact is required to devise control strategies for regional and global air pollution. PMID- 12552099 TI - Two Hsp70 family members expressed in atherosclerotic lesions. AB - Gene expression profiling was carried out comparing Con A elicited peritoneal macrophages from C57BL6 and FVBN wild-type and apolipoprotein (apo)E knockout mice. An EST, was expressed at higher levels in C57BL6 compared with FVBN mice. mapped to an atherosclerosis susceptibility locus on chromosome 19 revealed in an intercross between atherosclerosis-susceptible C57BL6 and atherosclerosis resistant FVBN apoE knockout mice. A combination of database search and Northern analysis confirmed that corresponded to 3'-UTR of a hitherto predicted gene, named HspA12A. Blasting the National Center for Biotechnology Information database revealed a closely related homologue, HspA12B. HspA12A and -B have very close human homologues. TaqMan analysis confirmed the increased HspA12A expression (2.6-fold) in elicited peritoneal macrophages from C57BL6 compared with FVBN mice. TaqMan analysis also revealed increased HspA12A and HspA12B expression (87- and 6-fold, respectively) in lesional versus nonlesional portions of the thoracic aorta from C57BL6 apoE knockout mice on a chow diet. In situ hybridization confirmed that both genes were expressed within lesions but not within nonlesional aortic tissue. Blasting of HspA12A and HspA12B against the National Center for Biotechnology Information database (NR) revealed a hit with the Conserved Domain database for Hsp70 (pfam00012.5, Hsp70). Both genes appear to contain an atypical Hsp70 ATPase domain. The BLAST search also revealed that both genes were more similar to primitive eukaryote and prokaryote than mammalian Hsp70s, making these two genes distant members of the mammalian Hsp70 family. In summary, we describe two genes that code for a subfamily of Hsp70 proteins that may be involved in atherosclerosis susceptibility. PMID- 12552101 TI - Crowding-dependent production of colony-stimulating factors by cultured syngeneic or allogeneic hematopoietic cells. AB - Mitogenic stimulation in vitro of mouse T lymphocytes induces the production of the hematopoietic cytokines granulocyte-macrophage colony-stimulating factor and IL-3. The present experiments showed that simple crowding of murine spleen or lymph node cells was a sufficient inducing stimulus. Crowding did not have this consequence for thymus or marrow cells or spleen cells from nu/nu or Rag-1-/- mice lacking T lymphocytes. Crowding for as short a period as 24 h was sufficient to allow subsequent cytokine production in sparse cultures. Purified T lymphocytes also exhibited low levels of crowding induction of cytokine production and cytokine production was enhanced by IL-2 and IFN-gamma. However, IFN-gamma-/- spleen cells exhibited similar crowding-induced colony-stimulating factor production to that of control spleen cells. Excess cell crowding inhibited cytokine production. This inhibition was not caused by receptor internalization of cytokines but may contribute to the failure to observe IL-3 production in lymphoid organs in vivo. Coculture of allogeneic spleen or peritoneal cells was a strong inducing signal for colony-stimulating factor production but this parameter was unable to detect autoreactivity of T lymphocytes in mice that lack suppressor of cytokine signaling 1 and exhibit T lymphocyte activation. PMID- 12552102 TI - Depletion of a single nucleoporin, Nup107, prevents the assembly of a subset of nucleoporins into the nuclear pore complex. AB - The nuclear pore complex (NPC) is a protein assembly that contains several distinct subcomplexes. The mammalian nucleoporin (Nup)-107 is part of a hetero oligomeric complex, that also contains Nup160, Nup133, Nup96, and the mammalian homolog of yeast Sec13p. We used transfection of HeLa cells with small interfering RNAs to specifically deplete mRNA for Nup107. In a domino effect, Nup107 depletion caused codepletion of a subset of other Nups on their protein but not on their mRNA level. Among the affected Nups was a member of the Nup107 subcomplex, Nup133, whereas two other tested members of this complex, Nup96 and Sec13, were unaffected and assembled into Nup107Nup133-deficient NPCs. We also tested several phenylalanine-glycine repeat-containing Nups that serve as docking sites for karyopherins. Some of these, such as Nup358, Nup214 on the cytoplasmic, and Nup153 on the nucleoplasmic side of the NPC, failed to assemble into Nup107Nup133-depleted NPCs, whereas p62, a Nup at the center of the NPC, was unaffected. Interestingly, the filamentous, NPC-associated protein Tpr also failed to assemble into the NPCs of Nup107-depleted cells. These data indicate that Nup107 functions as a keystone Nup that is required for the assembly of a subset of Nups into the NPC. Despite the depletion of Nup107 and the accompanying effects on other Nups, there was no significant effect on the growth rate of these cells and only a partial inhibition of mRNA export. These data indicate redundancy of Nups in the function of the mammalian NPC. PMID- 12552103 TI - Microbe forensics: oxygen and hydrogen stable isotope ratios in Bacillus subtilis cells and spores. AB - Bacillus subtilis, a Gram-positive, endospore-forming soil bacterium, was grown in media made with water of varying oxygen (delta(18)O) and hydrogen (deltaD) stable isotope ratios. Logarithmically growing cells and spores were each harvested from the cultures and their delta(18)O and deltaD values determined. Oxygen and hydrogen stable isotope ratios of organic matter were linearly related with those of the media water. We used the relationships determined in these experiments to calculate the effective whole-cell fractionation factors between water and organic matter for B. subtilis. We then predicted the delta(18)O and deltaD values of spores produced in nutritionally identical media and local water sources for five different locations around the United States. Each of the measured delta(18)O and deltaD values of the spores matched the predicted values within a 95% confidence interval, indicating that stable isotope ratio analyses may be a powerful tool for tracing the geographic point-of-origin for microbial products. PMID- 12552105 TI - Coarse-grained stochastic processes for microscopic lattice systems. AB - Diverse scientific disciplines ranging from materials science to catalysis to biomolecular dynamics to climate modeling involve nonlinear interactions across a large range of physically significant length scales. Here a class of coarse grained stochastic processes and corresponding Monte Carlo simulation methods, describing computationally feasible mesoscopic length scales, are derived directly from microscopic lattice systems. It is demonstrated below that the coarse-grained stochastic models can capture large-scale structures while retaining significant microscopic information. The requirement of detailed balance is used as a systematic design principle to guarantee correct noise fluctuations for the coarse-grained model. The coarse-grained stochastic algorithms provide large computational savings without increasing programming complexity or computer time per executive event compared to microscopic Monte Carlo simulations. PMID- 12552104 TI - Distinct gene expression profiles characterize the histopathological stages of disease in Helicobacter-induced mucosa-associated lymphoid tissue lymphoma. AB - Long-term colonization of humans with Helicobacter pylori can cause the development of gastric B cell mucosa-associated lymphoid tissue lymphoma, yet little is known about the sequence of molecular steps that accompany disease progression. We used microarray analysis and laser microdissection to identify gene expression profiles characteristic and predictive of the various histopathological stages in a mouse model of the disease. The initial step in lymphoma development is marked by infiltration of reactive lymphocytes into the stomach and the launching of a mucosal immune response. Our analysis uncovered molecular markers of both of these processes, including genes coding for the immunoglobulins and the small proline-rich protein Sprr 2A. The subsequent step is characterized histologically by the antigen-driven proliferation and aggregation of B cells and the gradual appearance of lymphoepithelial lesions. In tissues of this stage, we observed increased expression of genes previously associated with malignancy, including the laminin receptor-1 and the multidrug resistance channel MDR-1. Finally, we found that the transition to destructive lymphoepithelial lesions and malignant lymphoma is marked by an increase in transcription of a single gene encoding calgranulin AMrp-8. PMID- 12552106 TI - Molecular features of adult mouse small intestinal epithelial progenitors. AB - The adult mouse small intestinal epithelium undergoes perpetual regeneration, fueled by a population of multipotential stem cells and oligopotential daughters located at the base of crypts of Lieberkuhn. Although the morphologic features of small intestinal epithelial progenitors (SiEPs) are known, their molecular features are poorly defined. Previous impediments to purification and molecular characterization of SiEPs include lack of ex vivo clonigenic assays and the difficulty of physically retrieving them from their niche where they are interspersed between their numerous differentiated Paneth cell daughters. To overcome these obstacles, we used germ-free transgenic mice lacking Paneth cells to obtain a consolidated population of SiEPs with normal proliferative activity. These cells were harvested by laser capture microdissection. Functional genomics analysis identified 163 transcripts enriched in SiEPs compared with Paneth cell dominated normal crypt base epithelium. The dataset was validated by (i) correlation with the organellar composition of SiEPs versus Paneth cells, (ii) similarities to databases generated from recent mouse hematopoietic and neural stem cell genome anatomy projects, and (iii) laser capture microdissectionreal time quantitative RT-PCR studies of progenitor cell-containing populations retrieved from the small intestines, colons, and stomachs of conventionally raised mice. The SiEP profile has prominent representation of genes involved in c myc signaling and in the processing, localization, and translation of mRNAs. This dataset, together with our recent analysis of gene expression in the gastric stem cell niche, discloses a set of molecular features shared by adult mouse gut epithelial progenitors. PMID- 12552107 TI - One scorpion, two venoms: prevenom of Parabuthus transvaalicus acts as an alternative type of venom with distinct mechanism of action. AB - Scorpion venom is a complex mixture of salts, small molecules, peptides, and proteins. Scorpions employ this valuable tool in several sophisticated ways for subduing prey, deterring predators, and possibly during mating. Here, a subtle but clever strategy of venom utilization by scorpions is reported. Scorpions secrete a small quantity of transparent venom when initially stimulated that we propose to name prevenom. If secretion continues, a cloudy and dense venom that is white in color is subsequently released. The prevenom contains a combination of high K(+) salt and several peptides including some that block rectifying K(+) channels and elicit significant pain and toxicity because of a massive local depolarization. The presence of high extracellular K(+) in the prevenom can depolarize cells and also decrease the local electrochemical gradient making it more difficult to reestablish the resting potential. When this positive change to the K(+) equilibrium potential is combined with the blockage of rectifying K(+) channels, this further delays the recovery of the resting potential, causing a prolonged effect. We propose that the prevenom of scorpions is used as a highly efficacious predator deterrent and for immobilizing small prey while conserving metabolically expensive venom until a certain level of stimuli is reached, after which the venom is secreted. PMID- 12552109 TI - A general method for gene knockdown in mice by using lentiviral vectors expressing small interfering RNA. AB - We describe the use of lentiviral vectors expressing small interfering RNAs (siRNAs) to knock down the expression of specific genes in vitro and in vivo. A lentiviral vector capable of generating siRNA specific for GFP after transduction of 293T-GFP cell lines showed no GFP fluorescence. Furthermore, no GFP-specific RNA could be detected. When eggs from GFP-positive transgenic mice were transduced with lentivirus-expressing siGFP virus, reduced fluorescence could be seen in blastocysts. More interestingly, pups from F(1) progeny, which expressed siGFP, showed considerably diminished fluorescence and decreased GFP. We propose that an approach of combining transgenesis by lentiviral vectors expressing siRNAs can be used successfully to generate a large number of mice in which the expression of a specific gene(s) can be down-regulated substantially. We believe that this approach of generating "knockdown" mice will aid in functional genomics. PMID- 12552108 TI - Expanding expression of the 5-lipoxygenase pathway within the arterial wall during human atherogenesis. AB - Oxidation products of low-density lipoproteins have been suggested to promote inflammation during atherogenesis, and reticulocyte-type 15-lipoxygenase has been implicated to mediate this oxidation. In addition, the 5-lipoxygenase cascade leads to formation of leukotrienes, which exhibit strong proinflammatory activities in cardiovascular tissues. Here, we studied both lipoxygenase pathways in human atherosclerosis. The 5-lipoxygenase pathway was abundantly expressed in arterial walls of patients afflicted with various lesion stages of atherosclerosis of the aorta and of coronary and carotid arteries. 5-lipoxygenase localized to macrophages, dendritic cells, foam cells, mast cells, and neutrophilic granulocytes, and the number of 5-lipoxygenase expressing cells markedly increased in advanced lesions. By contrast, reticulocyte-type 15 lipoxygenase was expressed at levels that were several orders of magnitude lower than 5-lipoxygenase in both normal and diseased arteries, and its expression could not be related to lesion pathology. Our data support a model of atherogenesis in which 5-lipoxygenase cascade-dependent inflammatory circuits consisting of several leukocyte lineages and arterial wall cells evolve within the blood vessel wall during critical stages of lesion development. They raise the possibility that antileukotriene drugs may be an effective treatment regimen in late-stage disease. PMID- 12552110 TI - Secondary loss of chloroplasts in trypanosomes. PMID- 12552111 TI - Conformational dynamics of the Na+/K+-ATPase probed by voltage clamp fluorometry. AB - The method of voltage clamp fluorometry combined with site-directed fluorescence labeling was used to detect local protein motions of the fully active Na(+)K(+) ATPase in real time under physiological conditions. Because helix M5 extends from the cytoplasmic site of ATP hydrolysis into the cation binding region, we chose the extracellular M5-M6 loop of the sheep alpha(1)-subunit for the insertion of cysteine residues to identify reporter positions for conformational rearrangements during the catalytic cycle. After expression of the single cysteine mutants in Xenopus oocytes and covalent attachment of tetramethylrhodamine-6-maleimide, only mutant N790C reported molecular rearrangements of the M5-M6 loop by showing large, ouabain-sensitive fluorescence changes ( approximately 5%) on addition of extracellular K(+). When the enzyme was subjected to voltage jumps under Na(+)Na(+)-exchange conditions, we observed fluorescence changes that directly correlated to transient charge movements originating from the E(1)P-E(2)P transition of the transport cycle. The voltage jump-induced fluorescence changes and transient currents were abolished after replacement of Na(+) by tetraethylammonium or on addition of ouabain, showing that conformational flexibility is impaired under these conditions. Voltage dependent fluorescence changes could also be observed in the presence of subsaturating K(+) concentrations. This allowed to monitor the time course of voltage-dependent relaxations into a new stationary distribution of states under turnover conditions, showing the acceleration of relaxation kinetics with increasing K(+) concentrations. As a result, the stationary distribution between E(1) and E(2) states and voltage-dependent relaxation times can be determined at any time and membrane potential under Na(+)Na(+) exchange as well as Na(+)K(+) turnover conditions. PMID- 12552112 TI - Structural evidence for substrate strain in antibody catalysis. AB - The crystal structure of the Michaelis complex between the Fab fragment of ferrochelatase antibody 7G12 and its substrate mesoporphyrin has been solved to 2.6-A resolution. The antibody-bound mesoporphyrin clearly adopts a nonplanar conformation and reveals that the antibody catalyzes the porphyrin metallation reaction by straining/distorting the bound substrate toward the transition-state configuration. The crystal structures of the Fab fragment of the germ-line precursor antibody to 7G12 and its complex with the hapten N-methylmesoporphyrin have also been solved. A comparison of these structures with the corresponding structures of the affinity-matured antibody 7G12 reveals the molecular mechanism by which the immune system evolves binding energy to catalyze this reaction. PMID- 12552113 TI - Lower blood glucose, hyperglucagonemia, and pancreatic alpha cell hyperplasia in glucagon receptor knockout mice. AB - Glucagon, the counter-regulatory hormone to insulin, is secreted from pancreatic alpha cells in response to low blood glucose. To examine the role of glucagon in glucose homeostasis, mice were generated with a null mutation of the glucagon receptor (Gcgr(-/-)). These mice display lower blood glucose levels throughout the day and improved glucose tolerance but similar insulin levels compared with control animals. Gcgr(-/-) mice displayed supraphysiological glucagon levels associated with postnatal enlargement of the pancreas and hyperplasia of islets due predominantly to alpha cell, and to a lesser extent, delta cell proliferation. In addition, increased proglucagon expression and processing resulted in increased pancreatic glucogen-like peptide 1 (GLP-1) (1-37) and GLP-1 amide (1-36 amide) content and a 3- to 10-fold increase in circulating GLP-1 amide. Gcgr(-/-) mice also displayed reduced adiposity and leptin levels but normal body weight, food intake, and energy expenditure. These data indicate that glucagon is essential for maintenance of normal glycemia and postnatal regulation of islet and alpha and delta cell numbers. Furthermore, the lean phenotype of Gcgr(-/-) mice suggests glucagon action may be involved in the regulation of whole body composition. PMID- 12552114 TI - Origin-specific unwinding of herpes simplex virus 1 DNA by the viral UL9 and ICP8 proteins: visualization of a specific preunwinding complex. AB - Herpes simplex virus 1 contains three origins of replication; two copies of oriS and one of a similar sequence, oriL. Here, the combined action of multiple factors known or thought to influence the opening of oriS are examined. These include the viral origin-binding protein, UL9, and single-strand binding protein ICP8, host cell topoisomerase I, and superhelicity of the DNA template. By using electron microscopy, it was observed that when ICP8 and UL9 proteins were added together to oriS-containing supertwisted DNA, a discrete preunwinding complex was formed at oriS on 40% of the molecules, which was shown by double immunolabeling electron microscopy to contain both proteins. This complex was relatively stable to extreme dilution. Addition of ATP led to the efficient unwinding of approximately 50% of the DNA templates. Unwinding proceeded until the acquisition of a high level of positive supertwists in the remaining duplex DNA inhibited further unwinding. Addition of topoisomerase I allowed further unwinding, opening >1 kb of DNA around oriS. PMID- 12552115 TI - Diffusion-limited kinetics of the solution-solid phase transition of molecular substances. AB - For critical tests of whether diffusion-limited kinetics is an option for the solution-solid phase transition of molecular substances or whether they are determined exclusively by a transition state, we performed crystallization experiments with ferritin and apoferritin, a unique pair of proteins with identical shells but different molecular masses. We find that the kinetic coefficient for crystallization is identical (accuracy 200 kbp and consists of several hundred linear chromosomes that share elaborate terminal-specific sequence patterns. The highly peculiar organization of the ichthyosporean mtDNA raises questions about the mechanism of mitochondrial genome replication and chromosome segregation during cell division in this organism. Considering that the closest unicellular relatives of animals possess large, spacious, gene-rich mtDNAs, we posit that the distinct compaction characteristic of metazoan mitochondrial genomes occurred simultaneously with the emergence of a multicellular body plan in the animal lineage. PMID- 12552118 TI - Course of illness, hippocampal function, and hippocampal volume in major depression. AB - Studies have examined hippocampal function and volume in depressed subjects, but none have systematically compared never-treated first-episode patients with those who have had multiple episodes. We sought to compare hippocampal function, as assessed by performance on hippocampal-dependent recollection memory tests, and hippocampal volumes, as measured in a 1.5-T magnetic resonance imager, in depressed subjects experiencing a postpubertal onset of depression. Twenty never treated depressed subjects in a first episode of depression were compared with matched healthy control subjects. Seventeen depressed subjects with multiple past episodes of depression were also compared with matched healthy controls and to the first-episode patients. Both first- and multiple-episode depressed groups had hippocampal dysfunction apparent on several tests of recollection memory; only depressed subjects with multiple depressive episodes had hippocampal volume reductions. Curve-fitting analysis revealed a significant logarithmic association between illness duration and hippocampal volume. Reductions in hippocampal volume may not antedate illness onset, but volume may decrease at the greatest rate in the early years after illness onset. PMID- 12552119 TI - Chemogenomic identification of Ref-1/AP-1 as a therapeutic target for asthma. AB - Asthma is characterized by an oxidantantioxidant imbalance in the lungs leading to activation of redox-sensitive transcription factors, nuclear factor kappaB (NF kappaB), and activator protein-1 (AP-1). To develop therapeutic strategies for asthma, we used a chemogenomics approach to screen for small molecule inhibitor(s) of AP-1 transcription. We developed a beta-strand mimetic template that acts as a reversible inhibitor (pseudosubstrate) of redox proteins. This template incorporates an enedione moiety to trap reactive cysteine nucleophiles in the active sites of redox proteins. Specificity for individual redox factors was achieved through variations in X and Y functionality by using a combinatorial library approach. A limited array (2 x 6) was constructed where X was either NHCH(3) or NHCH(2) Ph and Y was methyl, phenyl, m-cyanophenyl, m-nitrophenyl, m acetylaniline, or m-methylbenzoate. These analogs were evaluated for their ability to inhibit transcription in transiently transfected human lung epithelial A549 cells from either an AP-1 or NF-kappaB reporter. A small-molecule inhibitor, PNRI-299, was identified that selectively inhibited AP-1 transcription (IC(50) of 20 microM) without affecting NF-kappaB transcription (up to 200 microM) or thioredoxin (up to 200 microM). The molecular target of PNRI-299 was determined to be the oxidoreductase, redox effector factor-1 by an affinity chromatography approach. The selective redox effector factor-1 inhibitor, PNRI-299, significantly reduced airway eosinophil infiltration, mucus hypersecretion, edema, and IL-4 levels in a mouse asthma model. These data validate AP-1 as an important therapeutic target in allergic airway inflammation. PMID- 12552120 TI - Dentate gyrus volume is reduced before onset of plaque formation in PDAPP mice: a magnetic resonance microscopy and stereologic analysis. AB - High-resolution magnetic resonance microscopy (MRM) was used to determine regional brain volumetric changes in a mouse model of Alzheimer's disease. These transgenic (Tg) mice overexpress human mutant amyloid precursor protein (APP) V717F under control of platelet-derived growth factor promoter (PDAPP mice), and cortical and hippocampal beta-amyloid (Abeta) deposits accumulate in heterozygotes after 8-10 mos. We used MRM to obtain 3D volumetric data on mouse brains imaged in their skulls to define genotype- and age-related changes. Hippocampal, cerebellar, and brain volumes and corpus callosum length were quantified in 40-, 100-, 365-, and 630-day-old mice. Measurements taken at age 100 days, before Abeta deposition, revealed a 12.3% reduction of hippocampus volume in Tg mice compared with WT controls. This reduction persisted without progression to age 21 mos. A significant 18% increase in hippocampal volume occurred between 40 and 630 days in WT mice, and no corresponding significant increase occurred in Tg mice. Cavalieri volume estimates of hippocampal subfields from 100-day-old Tg mice further localized a 28% volume deficit in the dentate gyrus. In addition, corpus callosum length was reduced by approximately 25% in Tg mice at all ages analyzed. In summary, reduced hippocampal volume and corpus callosum length can be detected by MRM before Abeta deposition. We conclude that overexpression of APP and amyloid may initiate pathologic changes before the appearance of plaques, suggesting novel targets for the treatment of Alzheimer's disease and further reinforcing the need for early diagnosis and treatment. PMID- 12552121 TI - Determination of the energetics governing the regulatory step in growth hormone induced receptor homodimerization. AB - Signaling in the human growth hormone (hGH)-human GH receptor system is initiated by a controlled sequential two-step hormone-induced dimerization of two hGH receptors via their extracellular domains (ECDs). Little is currently known about the energetics governing the important regulatory step in receptor signaling (step 2) because of previously existing experimental barriers in characterizing the binding of the second receptor (ECD2). A further complication is that ECD2 binds through contacts from two spatially distinct sites: through its N-terminal domain to hGH, and to ECD1 through its C-terminal domain, which forms a pseudo-2 fold symmetrical interaction between the stems of the two receptors. We report here a detailed evaluation of the energetics of step 2 binding using a modified surface plasmon resonance method that is able to measure accurately the kinetics of the trimolecular binding process and separate the effects of the two binding sites. The binding kinetics of 23 single and 126 ECD1-ECD2 pair-wise alanine mutations was measured. Although both of the ECD2 binding interfaces were found to be important, the ECD1-ECD2 stem-stem contact is the stronger of the two. It was determined that most residues in the binding interfaces act in additive fashion, and that the six residues common in both ECDs contribute very differently to homodimerization depending on which ECD they reside in. This interface is characterized by a binding "hot-spot" consisting of a core of three residues in ECD1 and two in ECD2. There is no similar hot-spot in the N-terminal domain of ECD2 binding to Site2 of hGH. This study suggests ways to engineer ECD molecules that will bind specifically to either Site1 or Site2 of hGH, providing novel reagents for biophysical and biological studies. PMID- 12552122 TI - Cells lying on a bed of microneedles: an approach to isolate mechanical force. AB - We describe an approach to manipulate and measure mechanical interactions between cells and their underlying substrates by using microfabricated arrays of elastomeric, microneedle-like posts. By controlling the geometry of the posts, we varied the compliance of the substrate while holding other surface properties constant. Cells attached to, spread across, and deflected multiple posts. The deflections of the posts occurred independently of neighboring posts and, therefore, directly reported the subcellular distribution of traction forces. We report two classes of force-supporting adhesions that exhibit distinct force-size relationships. Force increased with size of adhesions for adhesions larger than 1 microm(2), whereas no such correlation existed for smaller adhesions. By controlling cell adhesion on these micromechanical sensors, we showed that cell morphology regulates the magnitude of traction force generated by cells. Cells that were prevented from spreading and flattening against the substrate did not contract in response to stimulation by serum or lysophosphatidic acid, whereas spread cells did. Contractility in the unspread cells was rescued by expression of constitutively active RhoA. Together, these findings demonstrate a coordination of biochemical and mechanical signals to regulate cell adhesion and mechanics, and they introduce the use of arrays of mechanically isolated sensors to manipulate and measure the mechanical interactions of cells. PMID- 12552123 TI - Global variation in terrestrial conservation costs, conservation benefits, and unmet conservation needs. AB - Our ability to identify cost-efficient priorities for conserving biological diversity is limited by the scarcity of data on conservation costs, particularly at fine scales. Here we address this issue using data for 139 terrestrial programs worldwide. We find that the annual costs of effective field-based conservation vary enormously, across seven orders of magnitude, from <$0.1 to >$1,000,000 per km(2). This variation can be closely predicted from positive associations between costs per unit area and an array of indices of local development. Corresponding measures of conservation benefit are limited but show opposing global trends, being higher in less developed parts of the world. The benefit-to-cost ratio of conservation is thus far greater in less developed regions, yet these are where the shortfall in current conservation spending is most marked. Substantially increased investment in tropical conservation is therefore urgently required if opportunities for cost-effective action are not to be missed. PMID- 12552124 TI - Involvement of bone morphogenetic protein 4 (BMP-4) in pituitary prolactinoma pathogenesis through a Smad/estrogen receptor crosstalk. AB - Pituitary tumor development involves clonal expansion stimulated by hormones and growth factorscytokines. Using mRNA differential display, we found that the bone morphogenetic protein (BMP) inhibitor noggin is down-regulated in prolactinomas from dopamine D2-receptor-deficient mice. BMP-4 is overexpressed in prolactinomas taken from dopamine D2-receptor-deficient female mice, but expression of the highly homologous BMP-2 does not differ in normal pituitary tissue and prolactinomas. BMP-4 is overexpressed in other prolactinoma models, including estradiol-induced rat prolactinomas and human prolactinomas, compared with normal tissue and other pituitary adenoma types (Western blot analysis of 48 tumors). BMP-4 stimulates, and noggin blocks, cell proliferation and the expression of c Myc in human prolactinomas, whereas BMP-4 has no action in other human pituitary tumors. GH3 cells stably transfected with a dominant negative of Smad4 (Smad4dn; a BMP signal cotransducer) or noggin have reduced tumorigenicity in nude mice. Tumor growth recovered in vivo when the Smad4dn expression was lost, proving that BMP-4Smad4 are involved in tumor development in vivo. BMP-4 and estrogens act through overlapping intracellular signaling mechanisms on GH3 cell proliferation and c-myc expression: they had additive effects at low concentrations but not at saturating doses, and their action was inhibited by blocking either pathway with the reciprocal antagonist (i.e., BMP-4 with ICI 182780 or 17beta-estradiol with Smad4dn). Furthermore, coimmunoprecipitation studies demonstrate that under BMP-4 stimulation Smad4 and Smad1 physically interact with the estrogen receptor. This previously undescribed prolactinoma pathogenesis mechanism may participate in tumorigenicity in other cells where estrogens and the type beta transforming growth factor family have important roles. PMID- 12552125 TI - The critical regulator of embryonic hematopoiesis, SCL, is vital in the adult for megakaryopoiesis, erythropoiesis, and lineage choice in CFU-S12. AB - Gene targeting studies have shown that the transcription factor SCL is critically important for embryonic hematopoiesis, but the early lethality of SCL null mice has precluded the genetic analysis of its function in the adult. We have now generated a conditional knockout of SCL by using CreLox technology and an IFN inducible Cre transgenic mouse. Deletion of SCL in adult mice perturbed megakaryopoiesis and erythropoiesis with the loss of early progenitor cells in both lineages. This led to a blunted response to the hematopoietic stress induced by polyinosinic-polycytidylic acid, with a persistently low platelet count and hematocrit compared with controls. In contrast, progenitors of granulocyte and macrophage lineages were not affected, even in the setting of stress. Immature progenitor cells (day 12 colony-forming unit spleen) with multilineage capacity were still present in the SCL null bone marrow, but these progenitors had lost the capacity to generate erythroid and megakaryocyte cells, and colonies were composed of only myeloid cells. These results suggest that SCL is critical for megakaryopoiesis and erythropoiesis, but is dispensable for production of myeloid cells during adult hematopoiesis. PMID- 12552126 TI - A critical role for PPARalpha-mediated lipotoxicity in the pathogenesis of diabetic cardiomyopathy: modulation by dietary fat content. AB - To explore the role of peroxisome proliferator-activated receptor alpha (PPARalpha)-mediated derangements in myocardial metabolism in the pathogenesis of diabetic cardiomyopathy, insulinopenic mice with PPARalpha deficiency (PPARalpha( /-)) or cardiac-restricted overexpression [myosin heavy chain (MHC)-PPAR] were characterized. Whereas PPARalpha(-/-) mice were protected from the development of diabetes-induced cardiac hypertrophy, the combination of diabetes and the MHC PPAR genotype resulted in a more severe cardiomyopathic phenotype than either did alone. Cardiomyopathy in diabetic MHC-PPAR mice was accompanied by myocardial long-chain triglyceride accumulation. The cardiomyopathic phenotype was exacerbated in MHC-PPAR mice fed a diet enriched in triglyceride containing long chain fatty acid, an effect that was reversed by discontinuing the high-fat diet and absent in mice given a medium-chain triglyceride-enriched diet. Reactive oxygen intermediates were identified as candidate mediators of cardiomyopathic effects in MHC-PPAR mice. These results link dysregulation of the PPARalpha gene regulatory pathway to cardiac dysfunction in the diabetic and provide a rationale for serum lipid-lowering strategies in the treatment of diabetic cardiomyopathy. PMID- 12552127 TI - Release of C-type natriuretic peptide accounts for the biological activity of endothelium-derived hyperpolarizing factor. AB - Endothelial cells in most vascular beds release a factor that hyperpolarizes the underlying smooth muscle, produces vasodilatation, and plays a fundamental role in the regulation of local blood flow and systemic blood pressure. The identity of this endothelium-derived hyperpolarizing factor (EDHF), which is neither NO nor prostacyclin, remains obscure. Herein, we demonstrate that in mesenteric resistance arteries, release of C-type natriuretic peptide (CNP) accounts for the biological activity of EDHF. Both produce identical smooth muscle hyperpolarizations that are attenuated in the presence of high [K(+)], the G(i) G protein (G(i)) inhibitor pertussis toxin, the G protein-gated inwardly rectifying K(+) channel inhibitor tertiapin, and a combination of Ba(2+) (inwardly rectifying K(+) channel blocker) plus ouabain (Na(+)K(+)-ATPase inhibitor). Responses to EDHF and CNP are unaffected by the natriuretic peptide receptor (NPR)-AB antagonist HS-142-1, but mimicked by the selective NPR-C agonist, cANF(4 23). EDHF-dependent relaxation is concomitant with liberation of endothelial CNP; in the presence of the myoendothelial gap-junction inhibitor 18alpha glycyrrhetinic acid or after endothelial denudation, CNP release and EDHF responses are profoundly suppressed. These data demonstrate that acetylcholine evoked release of endothelial CNP activates NPR-C on vascular smooth muscle that via a G(i) coupling promotes Ba(2+)ouabain-sensitive hyperpolarization. Thus, we have revealed the identity of EDHF and established a pivotal role for endothelial derived CNP in the regulation of vascular tone and blood flow. PMID- 12552128 TI - Hg(II) binding to a weakly associated coiled coil nucleates an encoded metalloprotein fold: a kinetic analysis. AB - A detailed kinetic analysis of metal encapsulation by a de novo-designed protein is described. The kinetic mechanism of Hg(II) encapsulation in the three-stranded coiled coil formed by the peptide CH(3)CO-G LKALEEK CKALEEK LKALEEK G-NH(2) (Baby L9C) is derived by global analysis. The mechanism involves rapid initial collapse of two peptides by Hg(II) forming Hg(Baby L9C(-H))(2) with a linear thiolato Hg(II) bound to the cysteine sulfur atoms. Here, Baby L9C(-H) denotes Baby L9C with the cysteine thiol deprotonated. Addition of the third peptide, forming the three-stranded coiled coil, is the rate-determining step and results in an intermediate state involving two separate species. One of the species, termed the properly folded intermediate, undergoes rapid deprotonation of the third cysteine thiol, yielding the desired three-stranded coiled coil with an encapsulated trigonal thiolato Hg(II). The other species, termed the misfolded intermediate, rearranges in an experimentally distinguishable step to the properly folded intermediate. The order of the reaction involving the addition of the third peptide with respect to the concentration of Baby L9C indicates that addition of the third helix only occurs through reaction of Hg(Baby L9C(-H))(2) and Baby L9C that is unassociated with a coiled coil. Temperature dependence of the reaction afforded activation parameters for both the addition of the third helix (deltaH = 20(2) kcalmol; deltaS= 40(5) calmol K) and the rearrangement of the misfolded intermediate steps (deltaH = 23(2) kcalmol; deltaS= 27(5) calmol K). The mechanism is discussed with regard to metalloprotein folding and metalloprotein design. PMID- 12552129 TI - The genome sequence of Clostridium tetani, the causative agent of tetanus disease. AB - Tetanus disease is one of the most dramatic and globally prevalent diseases of humans and vertebrate animals, and has been reported for over 24 centuries. The manifestation of the disease, spastic paralysis, is caused by the second most poisonous substance known, the tetanus toxin, with a human lethal dose of approximately 1 ng/kg. Fortunately, this disease is successfully controlled through immunization with tetanus toxoid; nevertheless, according to the World Health Organization, an estimated 400,000 cases still occur each year, mainly of neonatal tetanus. The causative agent of tetanus disease is Clostridium tetani, an anaerobic spore-forming bacterium, whose natural habitat is soil, dust, and intestinal tracts of various animals. Here we report the complete genome sequence of toxigenic C. tetani E88, a variant of strain Massachusetts. The genome consists of a 2,799,250-bp chromosome encoding 2,372 ORFs. The tetanus toxin and a collagenase are encoded on a 74,082-bp plasmid, containing 61 ORFs. Additional virulence-related factors could be identified, such as an array of surface-layer and adhesion proteins (35 ORFs), some of them unique to C. tetani. Comparative genomics with the genomes of Clostridium perfringens, the causative agent of gas gangrene, and Clostridium acetobutylicum, a nonpathogenic solvent producer, revealed a remarkable capacity of C. tetani: The organism can rely on an extensive sodium ion bioenergetics. Additional candidate genes involved in the establishment and maintenance of a pathogenic lifestyle of C. tetani are presented. PMID- 12552130 TI - Developmental loss of miniature N-methyl-D-aspartate receptor currents in NR2A knockout mice. AB - The N-methyl-d-aspartate (NMDA) glutamate receptor (NMDAR), long implicated in developmental plasticity, shows decay time kinetics that shorten postnatally as NR2A subunits are added to the receptor. Neither the mechanism nor immediate effect of this change is known. We studied developing NMDAR currents by using visual neurons in slices from NR2A knockout (NR2AKO) and WT mice. Both strains show increased dendritic levels of synaptic density scaffolding protein PSD-95 with age. Dendritic levels of NR2A increased at the same time in WT and immunoprecipitated with PSD-95. PSD-95NMDAR binding was significantly decreased in the NR2AKO. Moreover, NMDAR miniature currents (minis) were lost and rise times of NMDAR evoked currents increased in mutant mice. Age-matched WT cells showed NR2A-rich receptors predominating in minis, yet slow NR2B mediated currents persisted in evoked currents. Disrupting photoreceptor activation of retinal ganglion cells eliminated increases in PSD-95 and NR2A in superior collicular dendrites of WT mice and slowed the loss of miniature NMDAR currents in NR2AKOs. These data demonstrate that NMDARs that respond to single quantal events mature faster during development by expressing the NR2A subunit earlier than NMDARs that respond to evoked release. We hypothesize that NR2A-rich NMDARs may be localized to the center of developing synapses by an activity-dependent process that involves the targeting of PSD-95 to the postsynaptic density. Neonatal receptors become restricted to perisynpatic or extrasynaptic sites, where they participate primarily in evoked currents. PMID- 12552131 TI - Eye opening induces a rapid dendritic localization of PSD-95 in central visual neurons. AB - The membrane-associated guanylate kinase PSD-95 scaffolds N-methyl-d-aspartate receptors to cytoplasmic signaling molecules, and associates with other glutamate receptors at central synapses. However, regulation of PSD-95 in vivo is poorly understood. We provide evidence of an activity-dependent redistribution of PSD-95 to dendrites in central visual neurons that is tied to eye opening. Six hours after eye opening, increased dendritic PSD-95 coimmunoprecipitates with the same proportions of stargazin, increased proportions of the N-methyl-d-aspartate receptor subunit NR2A, and decreased proportions of NR2B. Sustained high levels of PSD-95 in dendrites are dependent on continued pattern vision in juvenile but not mature animals, suggesting that the stabilization of PSD-95 at synapses may be involved in the control of developmental plasticity. PMID- 12552132 TI - Plant-like traits associated with metabolism of Trypanosoma parasites. AB - Trypanosomatid parasites cause serious diseases among humans, livestock, and plants. They belong to the order of the Kinetoplastida and form, together with the Euglenida, the phylum Euglenozoa. Euglenoid algae possess plastids capable of photosynthesis, but plastids are unknown in trypanosomatids. Here we present molecular evidence that trypanosomatids possessed a plastid at some point in their evolutionary history. Extant trypanosomatid parasites, such as Trypanosoma and Leishmania, contain several "plant-like" genes encoding homologs of proteins found in either chloroplasts or the cytosol of plants and algae. The data suggest that kinetoplastids and euglenoids acquired plastids by endosymbiosis before their divergence and that the former lineage subsequently lost the organelle but retained numerous genes. Several of the proteins encoded by these genes are now, in the parasites, found inside highly specialized peroxisomes, called glycosomes, absent from all other eukaryotes, including euglenoids. PMID- 12552134 TI - Multiple mutations and cancer. AB - Most human tumors are highly heterogenous. We have hypothesized that this heterogeneity results from a mutator phenotype. Our premise is that normal mutation rates are insufficient to account for the multiple mutations found in human cancers, and, instead, that cancers must exhibit a mutator phenotype early during their evolution. Here, we examine the current status and implications of the mutator phenotype hypothesis for the prognosis, treatment, and prevention of human cancers. PMID- 12552133 TI - The secretory proprotein convertase neural apoptosis-regulated convertase 1 (NARC 1): liver regeneration and neuronal differentiation. AB - Seven secretory mammalian kexin-like subtilases have been identified that cleave a variety of precursor proteins at monobasic and dibasic residues. The recently characterized pyrolysin-like subtilase SKI-1 cleaves proproteins at nonbasic residues. In this work we describe the properties of a proteinase K-like subtilase, neural apoptosis-regulated convertase 1 (NARC-1), representing the ninth member of the secretory subtilase family. Biosynthetic and microsequencing analyses of WT and mutant enzyme revealed that human and mouse pro-NARC-1 are autocatalytically and intramolecularly processed into NARC-1 at the (Y,I)VV(V,L)(L,M) downward arrow motif, a site that is representative of its enzymic specificity. In vitro peptide processing studies andor Ala substitutions of the P1-P5 sites suggested that hydrophobicaliphatic residues are more critical at P1, P3, and P5 than at P2 or P4. NARC-1 expression is highest in neuroepithelioma SK-N-MCIXC, hepatic BRL-3A, and in colon carcinoma LoVo-C5 cell lines. In situ hybridization and Northern blot analyses of NARC-1 expression during development in the adult and after partial hepatectomy revealed that it is expressed in cells that have the capacity to proliferate and differentiate. These include hepatocytes, kidney mesenchymal cells, intestinal ileum, and colon epithelia as well as embryonic brain telencephalon neurons. Accordingly, transfection of NARC-1 in primary cultures of embryonic day 13.5 telencephalon cells led to enhanced recruitment of undifferentiated neural progenitor cells into the neuronal lineage, suggesting that NARC-1 is implicated in the differentiation of cortical neurons. PMID- 12552135 TI - Flexible lid to the p53-binding domain of human Mdm2: implications for p53 regulation. AB - The stabilization of p53 against Mdm2-mediated degradation is an important event in DNA damage response. Initial models of p53 stabilization focused on posttranslational modification of p53 that would disrupt the p53-Mdm2 interaction. The N-terminal regions of both p53 and Mdm2 are modified in vivo in response to cellular stress, suggesting that modifications to Mdm2 also may affect the p53-Mdm2 interaction. Our NMR studies of apo-Mdm2 have found that, in addition to Mdm2 residues 25-109 that form the well ordered p53-binding domain that was observed in the p52-Mdm2 complex, Mdm2 residues 16-24 form a lid that closes over the p53-binding site. The Mdm2 lid, which is strictly conserved in mammals, may help to stabilize apo-Mdm2. It also competes weakly with peptidic and nonpeptidic antagonists. Modifications to the Mdm2 lid may disrupt p53-Mdm2 binding leading to p53 stabilization. Mdm2 and Mdm4 possess nearly identical p53 binding domains but different lids suggesting that lid modifications may select for p53 binding. PMID- 12552136 TI - Placental mammal diversification and the Cretaceous-Tertiary boundary. AB - Competing hypotheses for the timing of the placental mammal radiation focus on whether extant placental orders originated and diversified before or after the Cretaceous-Tertiary (KT) boundary. Molecular studies that have addressed this issue suffer from single calibration points, unwarranted assumptions about the molecular clock, andor taxon sampling that lacks representatives of all placental orders. We investigated this problem using the largest available molecular data set for placental mammals, which includes segments of 19 nuclear and three mitochondrial genes for representatives of all extant placental orders. We used the ThorneKishino method, which permits simultaneous constraints from the fossil record and allows rates of molecular evolution to vary on different branches of a phylogenetic tree. Analyses that used different sets of fossil constraints, different priors for the base of Placentalia, and different data partitions all support interordinal divergences in the Cretaceous followed by intraordinal diversification mostly after the KT boundary. Four placental orders show intraordinal diversification that predates the KT boundary, but only by an average of 10 million years. In contrast to some molecular studies that date the rat-mouse split as old as 46 million years, our results show improved agreement with the fossil record and place this split at 16-23 million years. To test the hypothesis that molecular estimates of Cretaceous divergence times are an artifact of increased body size subsequent to the KT boundary, we also performed analyses with a "KT body size" taxon set. In these analyses, interordinal splits remained in the Cretaceous. PMID- 12552137 TI - Working memory and fear conditioning. AB - Previous studies of associative learning implicate higher-level cognitive processes in some forms of classical conditioning. An ongoing debate is concerned with the extent to which attention and awareness are necessary for trace but not delay eye-blink conditioning [Clark, R. E. & Squire, L. R. (1998) Science 280, 77 81; Lovibond, P. F. & Shanks, D. (2002) J. Exp. Psychol. Anim. Behav. Processes 28, 38-42]. In trace conditioning, a short interval is interposed between the termination of the conditioned stimulus (CS) and the onset of the unconditioned stimulus (US). In delay conditioning, the CS and US overlap. We here investigate the extent to which human classical fear conditioning depends on working memory. Subjects had to carry out an n-back task, requiring tracking an item 1 or 2 back in a sequentially presented list of numbers, while simultaneously being tested for their ability to associate auditory cues with shocks under a variety of conditions (single-cue versus differential; delay versus trace; no task versus 0 , 1-, and 2-back). Differential delay conditioning proved to be more resilient than differential trace conditioning but does show a reduction due to task interference similar in slope to that found in trace conditioning. Explicit knowledge of the stimulus contingency facilitates but does not guarantee trace conditioning. Only the single-cue delay protocol shows conditioning during the more difficult working memory task. Our findings suggest that the larger the cognitive demands on the system, the less likely conditioning occurs. A postexperimental questionnaire showed a positive correlation between conditioning and awareness for differential trace conditioning extinction. PMID- 12552138 TI - The dichotomy of complex I: a sodium ion pump or a proton pump. PMID- 12552139 TI - Identification of nucleocytoplasmic cycling as a remote sensor in cellular signaling by databased modeling. AB - Considerable progress has been made in identifying the molecular composition of complex signaling networks controlling cell proliferation, differentiation, and survival. However, to discover general building principles and predict the dynamic behavior of signaling networks, it is necessary to develop quantitative models based on experimental observations. Here we report a mathematical model of the core module of the Janus family of kinases (JAK)-signal transducer and activator of transcription (STAT) signaling pathway based on time-resolved measurements of receptor and STAT5 phosphorylation. Applying the fitted model, we can determine the quantitative behavior of STAT5 populations not accessible to experimental measurement. By in silico investigations, we identify the parameters of nuclear shuttling as the most sensitive to perturbations and verify experimentally the model prediction that inhibition of nuclear export results in a reduced transcriptional yield. The model reveals that STAT5 undergoes rapid nucleocytoplasmic cycles, continuously coupling receptor activation and target gene transcription, thereby forming a remote sensor between nucleus and receptor. Thus, dynamic modeling of signaling pathways can promote functional understanding at the systems level. PMID- 12552140 TI - PCP in 2003: rapid reviewing and rapid publication. PMID- 12552141 TI - Plant RelA/SpoT homolog confers salt tolerance in Escherichia coli and Saccharomyces cerevisiae. AB - To analyze the mechanisms of salt tolerance in the halophyte Suaeda japonica, Escherichia coli was used as a host organism to undertake functional screening of cDNAs encoding proteins that may play an important role for the salt-tolerance mechanisms. A transformant expressing RelA/SpoT homolog (Sj-RSH) was found to have enhanced salt tolerance. In E. coli, RelA/SpoT controlled the amount of guanosine tetraphosphate (ppGpp) and guanosine pentaphosphate (pppGpp), which are the effectors of the bacterial stringent response. Complementation analysis using the relA mutant of E. coli showed that Sj-RSH conferred the phenotype associated with (p)ppGpp synthesis. Furthermore, expression of Sj-RSH driven by the GAL1 promoter also gave rise to enhanced salt tolerance in yeast. Northern blot analyses of the yeast transformant revealed that the transcriptional levels of stress responsive genes including GPD1, VMA6, BMH1, HYP1 and HOG1 were clearly enhanced in the Sj-RSH transformant when compared with an empty vector transformant under stress-free and 1.5 M NaCl stress conditions. These results suggest that (p)ppGpp synthesis mediated by plant RelA/SpoT homologs plays a critical role for the transcriptional induction of several stress responsive genes, directly or indirectly in yeast, and that the conserved stress-resistance system may exist in higher plants. PMID- 12552142 TI - CIP353 encodes an AP2/ERF-domain protein in potato (Solanum tuberosum L.) and responds slowly to cold stress. AB - We isolated the CIP353 cDNA, which encodes a novel cold-inducible protein, from cold-stored tubers of potato (Solanum tuberosum L.). The level of CIP353 transcripts began to increase in tubers 2 weeks after storage at 3 degrees C and continued increasing for at least 3 months during storage. This increase was not observed in tubers stored at >or=9 degrees C. The increased level of transcripts in tubers stored at 3 or 6 degrees C decreased when the tubers were shifted to 20 degrees C. These data suggest that CIP353 is a temperature-dependent and slowly responsive cold-inducible gene of potato. The middle of the deduced amino acid sequence of CIP353 cDNA showed high similarity to the AP2/ERF domain, which occurs in some plant regulatory factors. The deduced protein contained a putative basic nuclear-localization signal and potential acidic activation domains. These data suggest that CIP353 protein is a transcription factor of genes expressed in tubers under long-term storage at low temperatures. PMID- 12552143 TI - Two azuki bean XTH genes, VaXTH1 and VaXTH2, with similar tissue-specific expression profiles, are differently regulated by auxin. AB - To gain insight into the functional diversity of the XTH (xyloglucan endotransglucosylase/hydrolase) gene family, we analyzed the expression profiles of two azuki bean genes, VaXTH1 and VaXTH2, which share a striking resemblance in their amino acid sequences. The two XTH genes exhibit essentially similar tissue specific expression profiles, in that both mRNAs are found predominantly in the phloem fibers of growing internodes. However, their expression profiles are not identical. Whereas VaXTH1 is expressed in xylem cells in the basal part of the internode, little or no expression of VaXTH2 is found in the xylem. Furthermore, they exhibit spatially divergent RNA distribution profiles along the internode, VaXTH1 being expressed nearer to the top of the internode than VaXTH2. This indicates their temporally divergent expression profiles during development of the phloem fiber. Indole-3-acetic acid (IAA) up-regulates both of the mRNA levels. However, this effect of IAA on the VaXTH1 gene is nullified in 0.25 M mannitol, which prevents cell expansion without affecting auxin action per se. In contrast, the IAA-induced up-regulation of the VaXTH2 gene is not affected by mannitol. Furthermore, fusicoccin, which promotes acidification and growth, up regulates VaXTH1 expression, but not VaXTH2 expression. Thus, the two XTH genes are committed to different steps of the cell wall dynamics in the same cell type at different stages of phloem fiber development, and are regulated by IAA in different ways. PMID- 12552144 TI - Photoperiodic regulation of a 24-kD dehydrin-like protein in red-osier dogwood (Cornus sericea L.) in relation to freeze-tolerance. AB - A predominant 24-kD dehydrin-like protein was previously found to fluctuate seasonally within red-osier dogwood (Cornus sericea L.) stems. The current study attempted to determine what environmental cues triggered the accumulation of the 24-kD protein and to assess its potential role in winter survival. Controlled photoperiod and field studies confirmed that photoperiod regulates a reduction of stem water content (SWC), freeze-tolerance enhancement and accumulation of the 24 kD protein. Diverse climatic ecotypes, which are known to respond to different critical photoperiods, displayed differential reduction of SWC and accumulation of the 24-kD protein. A time-course study confirmed that prolonged exposure to short days is essential for SWC reduction, 24-kD protein accumulation, and freeze tolerance enhancement. Water deficit induced 24-kD protein accumulation and enhanced freeze-tolerance under long-day conditions. In all instances, freeze tolerance enhancement and 24-kD protein accumulation was preceded by a reduction of SWC. These results are consistent with the hypothesis that C. sericea responds to decreasing photoperiod, which triggers a reduction in SWC. Reduced SWC in turn may trigger the accumulation of the 24-kD protein and a parallel increase in freeze-tolerance. PMID- 12552145 TI - Endogenous alpha-ketol linolenic acid levels in short day-induced cotyledons are closely related to flower induction in Pharbitis nil. AB - Alpha-ketol linolenic acid [KODA, 9,10-ketol-octadecadienoic acid, that is 9 hydroxy-10-oxo-12(Z),15(Z)-octadecadienoic acid] is a signal compound found in Lemna paucicostata after exposure to stress, such as drought, heat or osmotic stress. KODA reacts with catecholamines to generate products that strongly induce flowering, although KODA itself is inactive [Yokoyama et al. (2000) Plant Cell Physiol. 41: 110; Yamaguchi et al. (2001) Plant Cell Physiol. 42: 1201]. We examined the role of KODA in the flower-induction process of Pharbitis nil (violet). KODA was identified for the first time in seedlings of P. nil grown under a flower-inductive condition (16-h dark exposure), by means of LC-SIM and LC-MS/MS. In addition, the changes in endogenous KODA levels (evaluated after esterification of KODA with 9-anthryldiazomethane) during the flower-inductive phase in short day-induced cotyledons were closely related to flower induction. The KODA concentration sharply increased in seedlings during the last 2 h of a 16 h dark period, while the KODA level showed no significant elevation under continuous light. The increase of KODA level occurred in cotyledonal blades, but not in other parts (petiole, hypocotyls and shoot tip). When the 16-h dark period was interrupted with a 10-min light exposure at the 8th h, flower induction was blocked and KODA level also failed to increase. The degree of elevation of KODA concentration in response to 16-h dark exposure was the highest when the cotyledons had just unfolded, and gradually decreased in seedlings grown under continuous light for longer periods, reaching the basal level at the 3rd day after unfolding. Flower-inducing ability also decreased in a similar manner. These results suggest that KODA may be involved in flower induction in P. nil. PMID- 12552146 TI - Plants impaired in state transitions can to a large degree compensate for their defect. AB - Arabidopsis thaliana plants lacking the PSI-H or PSI-L subunit of photosystem I have been shown to be severely affected in their ability to perform state transitions, but no visual phenotype was observed when these plants were grown under different light quantities and qualities. However, the chloroplasts in the PSI-H- and PSI-L-less plants contained fewer and more extended grana stacks. The plants lacking PSI-H or PSI-L were characterised with respect to their photosynthetic performance. Wild-type plants adjusted the non-photochemical fluorescence quenching to maintain constant levels of PSII quantum yield and reduction of the plastoquinone pool. In contrast, the plants deficient in state transitions had a more reduced plastoquinone pool and consequently, a less efficient PSII-photochemistry under growth-light conditions and in state 2. The maximal photosynthetic capacity and the quantum efficiency of oxygen evolution were diminished by 8-14% in the PSI-H-less plants. Under growth-light conditions, the stroma was similarly reduced in the PSI-H-less plants and the rate of cyclic electron transport was unchanged. Pigment analysis showed that the xanthophyll cycle was not upregulated in order to compensate for the lack of state transitions. In general, the plants lacking PSI-H and PSI-L showed a decreased ability to optimise photosynthesis according to the light conditions. PMID- 12552147 TI - Evidence for nucleotide-dependent passive H+ transport protein in the plasma membrane of barley roots. AB - Plasma membranes were isolated from barley roots by two-phase partitioning, and octylglucoside-soluble and -insoluble fractions were obtained. The insoluble fractions were reconstituted into liposomes, and the plasma membrane H(+)-ATPase was shown to participate in MgATP-dependent H(+) transport activity. The H(+) transport was decreased when the octylglucoside-soluble fraction was reconstituted together with the insoluble fraction. The decrease was not due to inhibition of the H(+)-ATPase, but rather was likely due to the increased H(+) leakage from the proteoliposome. The octylglucoside-soluble fraction was, therefore, reconstituted in the liposomes and the passive H(+) transport was determined using the pH jump method. A pH gradient across the membranes was generated by the pH jump, and the gradient was found to be dissipated by passive H(+) transport. The H(+) transport required ATP, K(+), and valinomycin. The H(+) transport also occurred when ADP, AMP, GTP, or ATP-gamma-S was present instead of ATP, and did not occur when the octylglucoside-soluble fraction was boiled before the reconstitution. These findings suggest that nucleotide-dependent H(+ )transport protein is present in the plasma membrane of root cells. PMID- 12552148 TI - Isolation and characterization of the ACC synthase genes from lettuce (Lactuca sativa L.), and the involvement in low pH-induced root hair initiation. AB - Root hair formation is induced when lettuce seedlings are transferred from pH 6.0 to pH 4.0. Ethylene, auxin and light are essential to this process. To investigate the role of ethylene in root hair initiation, we isolated two 1 aminocyclopropane-1-carboxylic acid (ACC) synthase genes (Ls-ACS1 and Ls-ACS2). Seven motifs of known ACS proteins were highly conserved in Ls-ACS1 and Ls-ACS2. The Ls-ACS1 and Ls-ACS2 mRNA levels were constant at pH 6.0, which were lower than that in seedlings at pH 4.0. Ls-ACS1 and Ls-ACS2 transcripts accumulated at pH 4.0 and reached peak levels at 1 h and 30 min after acidification, respectively. Indole-3-acetic acid (IAA) induced the accumulation of both Ls-ACS1 and Ls-ACS2 transcripts, whereas ACC induced only Ls-ACS1 mRNA. These results suggest that acidification-induced auxin accumulations increase the Ls-ACS2 levels, which together with Ls-ACS2-induced ethylene raise the levels of Ls-ACS1. Furthermore, blue and white light gave the highest levels of both Ls-ACS1 mRNA and ethylene production. Darkness was less effective, and red light had an intermediate effect. The different light conditions had no effect on the levels of Ls-ACS2 mRNA. These observations support the involvement of Ls-ACS1 in the production of ethylene, which is crucial for root hair initiation. PMID- 12552149 TI - Linear dominance relationship among four class-II S haplotypes in pollen is determined by the expression of SP11 in Brassica self-incompatibility. AB - Self-incompatibility (SI) prevents self-fertilization by rejecting pollen from plants with the same S phenotype. The Brassica SI system is controlled sporophytically by multiple alleles at the single locus, S, and dominance relationships among S haplotypes are observed in both stigma and pollen. We have identified previously five different class-II S haplotypes in Brassica campestris. Here, we performed test-crosses between S heterozygotes and their respective parental S homozygotes for four of these class-II S haplotypes, and observed a linear dominance relationship on the pollen side. To determine how this relationship is controlled, we performed RNA gel blot analyses for six S heterozygotes and their respective parental S homozygotes using the corresponding SP11 clone as a probe. In all six S heterozygotes, SP11 derived from a dominant haplotype was predominantly expressed, and SP11 derived from a recessive haplotype was repressed. Thus, the linear dominance relationship of the SI phenotype on the pollen side is regulated by the expression of SP11. PMID- 12552150 TI - Binding and functional properties of the extrinsic proteins in oxygen-evolving photosystem II particle from a green alga, Chlamydomonas reinhardtii having his tagged CP47. AB - Oxygen-evolving photosystem II (PSII) particles were purified from Chlamydomonas reinhardtii having His-tag extension at the C terminus of the CP47 protein, by a single-step Ni(2+)-affinity column chromatography after solubilization of thylakoid membranes with sucrose monolaurate. The PSII particles consisted of, in addition to intrinsic proteins, three extrinsic proteins of 33, 23 and 17 kDa. The preparation showed a high oxygen-evolving activity of 2,300-2,500 micro mol O(2) (mg Chl)(-1) h(-1) in the presence of Ca(2+) using ferricyanide as the electron acceptor, while its activity was 680-720 micro mol O(2) (mg Chl)(-1) h( 1) in the absence of Ca(2+) and Cl(-) ions. The activity was 710-820 micro mol O(2) (mg Chl)(-1) h(-1) independent of the presence or absence of Ca(2+) and Cl( ) when 2,6-dichloro-p-benzoquinone was used as the acceptor. These activities were scarcely inhibited by DCMU. The kinetics of flash-induced fluorescence decay revealed that the electron transfer from Q(A)(-) to Q(B) was significantly inhibited, and the electron transfer from Q(A)(-) to ferricyanide was largely stimulated in the presence of Ca(2+). These results indicate that the acceptor side, Q(B) site, was altered in the PSII particles but its donor side remained intact. Release-reconstitution experiments revealed that the extrinsic 23 and 17 kDa proteins were released only partially by NaCl-wash, while most of the three extrinsic proteins were removed when treated with urea/NaCl, alkaline Tris or CaCl(2). The 23 and 17 kDa proteins directly bound to PSII independent of the other extrinsic proteins, and the 33 kDa protein functionally re-bound to CaCl(2) treated PSII which had been reconstituted with the 23 and 17 kDa proteins. These binding properties were largely different from those of the extrinsic proteins in higher plant PSII, and suggest that each of the three extrinsic proteins has their own binding sites independent of the others in the green algal PSII. PMID- 12552151 TI - CvADH1, a member of short-chain alcohol dehydrogenase family, is inducible by gibberellin and sucrose in developing watermelon seeds. AB - To understand the molecular mechanisms that control seed formation, we selected a seed-preferential gene (CvADH1) from the ESTs of developing watermelon seeds. RNA blot analysis and in situ localization showed that CvADH1 was preferentially expressed in the nucellar tissue. The CvADH1 protein shared about 50% homology with short-chain alcohol dehydrogenase including ABA2 in Arabidopsis thaliana, stem secoisolariciresinol dehydrogenase in Forsythia intermedia, and 3beta hydroxysterol dehydrogenase in Digitalis lanata. We investigated gene-expression levels in seeds from both normally pollinated fruits and those made parthenocarpic via N-(2-chloro-4-pyridyl)-N'-phenylurea treatment, the latter of which lack zygotic tissues. Whereas the transcripts of CvADH1 rapidly started to accumulate from about the pre-heart stage in normal seeds, they were not detectable in the parthenocarpic seeds. Treating the parthenogenic fruit with GA(3) strongly induced gene expression, up to the level accumulated in pollinated seeds. These results suggest that the CvADH1 gene is induced in maternal tissues by signals made in the zygotic tissues, and that gibberellin might be one of those signals. We also observed that CvADH1 expression was induced by sucrose in the parthenocarpic seeds. Therefore, we propose that the CvADH1 gene is inducible by gibberellin, and that sucrose plays an important role in the maternal tissues of watermelon during early seed development. PMID- 12552152 TI - The plastid clpP gene may not be essential for plant cell viability. AB - The plastid gene clpP is widely regarded as essential for chloroplast function and general plant cell survival. In this note we provide evidence that certain lines of non-photosynthetic maize (Zea mays) Black Mexican Sweet (BMS) suspension cells do not carry clpP in their plastid genomes. We also discuss several incidences in the literature where clpP is either missing or not expressed in other non-green cell lines and plants. We conclude that clpP is not required for general plant cell survival but instead may only be essential for the development and/or function of plastids with active gene expression. PMID- 12552153 TI - Chlorophyllase as a serine hydrolase: identification of a putative catalytic triad. AB - Chlorophyllases (Chlases), cloned so far, contain a lipase motif with the active serine residue of the catalytic triad of triglyceride lipases. Inhibitors specific for the catalytic serine residue in serine hydrolases, which include lipases effectively inhibited the activity of the recombinant Chenopodium album Chlase (CaCLH). From this evidence we assumed that the catalytic mechanism of hydrolysis by Chlase might be similar to those of serine hydrolases that have a catalytic triad composed of serine, histidine and aspartic acid in their active site. Thus, we introduced mutations into the putative catalytic residue (Ser162) and conserved amino acid residues (histidine, aspartic acid and cysteine) to generate recombinant CaCLH mutants. The three amino acid residues (Ser162, Asp191 and His262) essential for Chlase activity were identified. These results indicate that Chlase is a serine hydrolase and, by analogy with a plausible catalytic mechanism of serine hydrolases, we proposed a mechanism for hydrolysis catalyzed by Chlase. PMID- 12552154 TI - [Present-day approach to pharmacological and clinical aspects of novel antidepressants]. AB - Depression is the most common illness that affects a large number of individuals in all countries. Recent evidence suggest that depressive episodes if left untreated may heighten severity of subsequent episodes and may increase need for more health care resources. The first antidepressants, tricyclics and monoamine oxidase inhibitors, became available in the late 1950s. A progressive tightening of requirements by drug licensing authorities has ensured that efficacy evidence is good for most antidepressants that are in use. Contemporary antidepressant classification system is based on the mechanism of action, which is presumed to be responsible for their antidepressant effects. A pharmacodynamic system of classification has advantages because it incorporates the current theories of disease pathophysiology. Understanding the basic aspects of mechanism of action of antidepressants is important for treatment of depressive episode, for development of augmenting strategies and combining antidepressants with other antidepressants or antipsychotics. Antidepressants as a class of psychotropic medication have the broad range of indications. The choice of initial antidepressant legitimately varies considerably among clinicians and countries. Referring to some differences of recommendations for the first line treatment of depressive episode we suppose that the choice of antidepressant medication must be individualized for a particular patient. Novel antidepressants (SSRI, SNRI, NaSSA, NARI, NDRI and other) are safe and better tolerated. Metabolism of novel antidepressants is much improved compared with MAOIs and TCAs. The combination of antidepressants is an important clinical issue. There are the following principles of combining antidepressants: 1. to combine mechanisms of action not just drugs, 2. to combine antidepressants and to promote pharmacological synergy and tolerability, 3. to use important synergies within the serotonin, noradrenaline and even dopamine monoaminergic systems. Adequate treatment of depression including modern treatment approaches has the potential to reduce suffering and disability substantially and minimise the risk of suicide. PMID- 12552155 TI - [Clinical characteristics and possibilities of early diagnosis of Chlamydia trachomatis infection in adolescent girls]. AB - Genital Chlamydia trachomatis infection is the most frequently found sexually transmitted disease among sexually active adolescent girls aged 15-19 years. In general, C. trachomatis infection tends to be less abrupt in onset, clinical course is usually asymptomatic or with mild symptoms. Infection results in changes of inner genitalia and can be the reason of serious late complications (pelvic inflammatory disease and infertility). Purpose of the study was to evaluate complaints, clinical peculiarities, results of gynecologic examination and possibilities of early diagnosis of C. trachomatis infection among adolescent girls. METHODS: Anonymous questionnaire was used in the study, in which 487 girls aged 15-19 years participated. All 240 sexually active girls were advised to perform tests for STD in Kaunas University of Medicine Hospital. Gynecologic examination, CT/NG DNA hybridisation test from vagina and PCR from cervix for detection C. trachomatis infection were performed. RESULTS: Eighteen percent of examined girls were infected with C. trachomatis. Complaints of fool odor discharge from vagina, non-specific changes of vagina and cervix let us suspect for C. trachomatis infection with statistical significance. CONCLUSIONS: The high prevalence of C. trachomatis infection among adolescent girls was found; the C. trachomatis infection could be suspected with non-specific changes of vagina and cervix and complaints of fool odor discharge from vagina, which helps us to prescribe early and adequate treatment. PMID- 12552156 TI - [Analysis of burns' causes of children admitted to the National Burns Center in 1995-2000]. AB - A retrospective multifactorial epidemiological study of pediatric patients admitted to the National Burns Center was undertaken, since January 1995 to December 2000. Nine hundred sixty-nine patients were admitted. The objective of this study was to describe the burns characteristics of different pediatric age groups. Scalds were the main cause of burns in the youngest group - less than 7 year age, vs. flame, which was one of the mains causes of burn in the elder group. The home was the most common site of injury - 93.4%. The mean patient age was 3.43+/-0.12 (range from 6 months to 16 years). The mean percentage body surface area burned was 8.87+/-0.28 percent. PMID- 12552157 TI - [Diagnostic value of pleural fluid cytologic examination]. AB - The study included 200 patients with pleural effusion. Pleural effusions were transudative in 48 (24%) and exudative in 152 (76%) of cases. Congestive cardiac failure (14.5%), nephrotic syndrome (5.5%), and liver cirrhosis (2.5%) were the most common etiological diagnoses of transudate cases. Malignant effusion (16.5%), pneumonia (13%), pleural empyema (9%), tuberculosis (6%), and pulmonary embolism (5.5%) were the most common etiological diagnoses of exudative cases. Thirty-two (16%) cases of exudative pleural effusions were of undertermined etiology. Polymorphonuclear leucocytes predominated in 48 patients with exudative pleural effusions. The most common etiological diagnoses were pneumonia (41.67%), pleural empyema (39.59%) and pulmonary embolism (10.42%). Lymphocytes in pleural fluid were predominant in 63 patients, with malignant (6.34%), tuberculous pleurisy (19.02%), pulmonary embolism (6.34%), trauma (6.34%), and (46.11%) cases in patients with pleural exudate undertermined etiology. Eosinophyls were predominant in 16 (8%) patients with exudative pleural effusions. The most common etiology of eosinophilic pleural fluid were pneumonia (37.5%), malignant pleural effusion (25%), pulmonary embolism (12.5%), pyopneumothorax (6.25%) and trauma 6.25%. From 16 patients with eosinophilic pleural exudate, in 31% cases air, in 12.5% blood in pleural fluid were determined and in 12.5% cases previous pleural puncture was performed. Pleural fluid eosinophilia is most commonly associated with the presence of air or blood in the pleural fluid (correalation index 0.82). Malignant pleural effusions were determined in 33 patients. Malignant cells in pleural fluid were identified in 25 cases. The diagnostic sensitivity of pleural fluid cytology for malignant pleural effusions were 76%. Hemoragic pleuritis was determined in 18 and hemothorax in 4 patients. Etiology of hemothorax were trauma (75%) and coaguliopathia (25%). Most common etiological diagnoses of hemoragic pleuritis were neoplasia (33.3%), pulmonary embolism (16.65%), trauma (16.65%), pneumonia (11.11%), and congestive cardiac failure (11.11%). Diagnostic sensitivity and specifity of hemoragic pleuritis is low, 58% and 45% respectively. PMID- 12552158 TI - [Clinical analysis of the patients with mumps]. AB - Mump morbidity in Lithuania and the City of Siauliai has rapidly increased in 1999; 5946 and 879, respectively. During the past decade, the number of cases was much lower: in Lithuania from 383 to 632, and in the City of Siauliai from 6 to 31. There were 87 cases of mumps diagnosed (M-72, F-15). The age of patients was 15-42 years. Most patients were 15-20 years old. The majority of patients was admitted to the department in spring and summer. The most common acute symptoms of mumps were: fever (97.70%) and painful enlargement of the salivary glands (71.26%). Rare symptoms were: painful swallowing, weakness, nausea and vomiting. Orchitis was observed in 43 patients (59.72% of male) and increasing of amylases in 52 patients (59.77%). Meningitis developed in 15 patients while 1 of them had cochleoneuritis, which led to deafness. The duration of treatment in hospital was 3-39 days (avg. 8.29). Two pregnant women were treated with mumps in our department. The condition of their fetus was not affected. PMID- 12552159 TI - [Importance of voice quality evaluation in the assessment of treatment outcome after endolaryngeal microsurgery]. AB - The aim of the study was to compare subjective and quantitative voice characteristics changes after endolaryngeal microsurgery with respect to different postoperative follow-up term. We evaluated 80 surgically-treated patients aged 14-81 years (mean 42.6+/-13.8 years) with benign vocal cord (VC) lesions (polyps, n=37; cysts, n=16; Reinke's edema, n=18; papilomata, n=9). Patients were evaluated three times - before operation, less than two weeks (mean 1.0+/-0.4) following the operation and more than two weeks (mean 6.5+/-3.9) following the operation. Control group consisted of 122 healthy voice patients. Subjective voice evaluation composed of experts assessment (GRBAS hoarseness scale, visual analogue scale (VAS) of voice quality) and patients self-evaluation (VAS, voice handicap index, and the impression of what repercussions the voice problem has on professional life and emotions). Seven quantitative voice parameters obtained from voice range profile, laryngostroboscopy and registering of maximum phonation time were analyzed as well as overall vocal dysfunction degree (VDD). The data of second postoperative examination were statistically significantly better compared to the first postoperative follow-up (p<0.001). During the first postoperative examination the subjectively healthy voice (G=0) was found for 13.8% (11), quantitatively (VDD=0) for 22.5% (18) of patients, during the second - 63.8% (51) and 53.8% (43). The best results were for patients with vocal cords polyps. The complex voice assessment may be useful for evaluation of voice function recovery and efficacy of surgical treatment. PMID- 12552160 TI - [The European Registry of Cataract Surgery Outcomes: Clinic of Ophthalmology, Kaunas University of Medicine]. AB - PURPOSES: Evaluate the outcome of cataract surgery in Clinic of Ophthalmology, Kaunas University of Medicine (COKUM) and to compare it with the outcome of the European Cataract Outcome Study Group (ECOSG) data. METHODS: The study was started on the 1st of October, 2000 and ended on the 30th April, 2001 on the basis of the protocol of the European Cataract Outcome Study Group. Every patient at each participating unit having surgery during the first study month was evaluated. The study was closed 6 months after surgery. RESULTS: The study enroled 3944 patients, out of them 361 was from COKUM. The mean induced astigmatism was 0.86+/-0.21 D in COKUM and 0.63+/-0.23 D in ECOSG. The visual acuity of the operated eye was 0.3 or lower in 28.1 percent of patients, 0.4-0.7 in 31.7 percent and 0.8 or higher in 40.2 percent of COKUM patients in the whole study group percentages were 11.8, 27.6 and 60.6, respectively. 44.3 percent of COKUM patients; underwent phacoemulsiphication, while among ECOSG this procedure was the most common (91.8 percent). The number of complications during surgery was 5.5 percent of all cases in COKUM while in European countries it was 3.7 percent. CONCLUSION: Cataract surgery data collected from 39 units in 18 European countries allowed participants to compare their performance with that of colleagues in an anonymous manner. This study is also an indicator of cataract surgery development in COKUM and in Lithuania. PMID- 12552161 TI - [Hiatal hernia and gastroesophageal reflux: possibilities and results of surgical treatment]. AB - In this article we analyze our experience of surgical treatment of hiatal hernia, complicated with gastroesophageal reflux. We operated 134 patients with hernia hiatus esophagi, complicated with gastroesophageal reflux, from 03.1998 till 10.2001. One hundred twenty-six Nissen and 8 Toupet laparoscopic gastrofundoplications were performed. We evaluated clinical signs of gastroesophageal reflux, performed endoscopy and esophageal biopsy with histological examination and stomach X-ray examination with barium meal before the operation. Esophagus and stomach X-ray examination with water contrast on the first day after operation were performed in order to evaluate the position and function of created wrap. We also analyzed intraoperative and postoperative complications. Long-term follow-up (12 months) was obtained by using a structured questionnaire. We evaluated heartburn, dysphagia, regurgitation and patient's satisfaction of surgery. RESULTS. Postoperative complications rate was 8.96%. Eighty-two percent of our patients completed our questionnaire. Ninety-one percent of patients had no heartburn signs, 95.5% any signs of regurgitation. Eighty-three percent of our patients were satisfied with our performed laparoscopic gastrofundoplication. We performed 6 refundoplications, when gastroesophageal reflux clinical signs renewed shortly after operation. CONCLUSIONS. Laparoscopic gastrofundoplication is a safe and effective treatment of hernia hiatus esophagi, complicated with gastroesophageal reflux. Operation success was about 90% in our study. Recurrences are more frequent in elderly patients or those with long disease anamnesis. Refundoplications can be successfully done laparoscopicaly as well. PMID- 12552162 TI - [Variability and diagnosis of pulmonary tuberculosis]. AB - Five hundred-ninety new cases of tuberculosis were analyzed in Republic Santariskiu Tuberculosis and Lung Disease Hospital during 06 1997-06 2001. Mean age of patients - 47+/-30 years. All cases were classified based on international tuberculosis classification: pulmonary tuberculosis (A15, A16) - 409 cases; tuberculosis pleurisy (A15.6, A16.5) - 169; miliary tuberculosis (A19) - 11; generalized tuberculous lymphadenitis (A18.5) - 1. Pulmonary tuberculosis confirmed by sputum smear (A15.0) was prevalent in the structure of new tuberculosis cases (26.4%); pulmonary tuberculosis was confirmed by culture (A15.1) in 14.1%; by histology (A15.2) - in 10.3%. Unconfirmed pulmonary tuberculosis (A16) and tuberculous pleurisy (A16.5) amounted to 28.5%. Liver biopsy and positive sputum cultures for mycobacterias mostly confirmed tuberculosis diagnosis in miliary tuberculosis cases. From all tuberculous pleurisy cases in 65% the diagnosis was confirmed by bacteriology and/or histology. PMID- 12552163 TI - [Pharmacological cardioversion of atrial fibrillation with intravenous and oral propafenone]. AB - The aim of the study was to evaluate the efficacy of intravenous and oral propafenone for termination of recent onset (<48 hrs) atrial fibrillation. Forty seven nonvalvular patients with atrial fibrillation were included in the study. Sinus rhythm was restored in 19/31 (61.3%) patients after intravenous treatment: in 15/31 (48.4%) atrial fibrillation was stopped after bolus of propafenone in dose of 2 mg/kg body weight, in 4/16 (25.0%) - by followed infusion of medicine in dose of 0.008 mg/kg/min during 2 hrs. A single 600 mg oral dose of propafenone restored sinus rhythm in 9/16 (56.2%) patients during 4 hrs. Transformation of atrial fibrillation into regular atrial flutter was observed in 4 cases; in 1 of them right bundle branch block developed. PMID- 12552164 TI - [Case of herpes gestationis in 5th pregnancy]. AB - We analyze the case of herpes gestationis to 36 year-old woman, which started on 24th week of 5th gestation. During the past gestations there were no fillings. Treatment with prednisolon 40 mg per day only reduced the signs. After the birth treatment with dapson 100 mg per day for the 5-days period gave a visible effect quickly. The treatment was completed 3 months ago and now there are no signs of illness. PMID- 12552165 TI - [Technology and analysis of Ginkgo tincture]. AB - The article deals with the production of the tincture of Ginkgo leaves while selecting extractant, the size of particles in a raw material, and the method of extraction. The optimal concentration of the extract was established by experiment and was the following: ethanol of 70 percent (V/V), the particles size 2-3 mm, the production method percolation, and the flow speed of tincture 0.5 ml/min. The tinctura was analyzed at determination of the sum of flavonoids in terms of quercetin, dry residue, the concentration of ethanol, density, refraction index, heavy metals and microbe pollution. The stability of the tincture and its expiry date were fixed. PMID- 12552166 TI - [Influence of lifestyle, biological and other factors on subjective evaluation of physical functioning]. AB - OBJECTIVE: To assess the strength of relationship between various lifestyle, biological, psychosocial factors, chronic conditions, pain and subjective physical performance, evaluated by "Physical functioning" scale from "Rand-36 Health Survey" questionnaire. MATERIAL AND METHODS: We used a standardized generic health related quality of life questionnaire. The study population was based on the original sample, formerly explored during the study, carried out by the Department of Propedeutics to Internal Medicine. It enroled 575 primary care patients from seven small towns located in Kaunas region. The effect exerted by various factors on ratings of subjective physical performance was evaluated by means of multiple logistic regression. Ratings of the "Pain" scale, as well as subjective rating of change in health during preceding year were included in the analysis. RESULTS: The biggest influence on rating of physical functioning was exerted by reported deterioration in health during preceding year and severity of pain. Among biological factors, age, diastolic blood pressure (>80 mmHg), body mass index (> or =35) and FEV1 (%) were adversely related to subjective ratings of physical health. Smoking status, chronic respiratory conditions, disablement group, perceived lack of social support, reported shortage of food expenditures and being pensioned off, all remained significant predictors of worsened physical functioning. CONCLUSIONS: Perceived physical health is affected by both objective biological and subjective factors related to the history of disease as well as psychosocial factors. These should be taken into account when planning clinical trials where health-related quality of life is intended to be analyzed among outcomes of interest. PMID- 12552167 TI - [Professional associations of pharmacists in Lithuania: history and directions of activity]. AB - The aim of this study was to describe the historical development of pharmaceutical organisations, their tasks and ability to fulfill them. In the XIXth century a rapid development of the science of pharmacy influenced a need for scientific collaboration. Thus, in 1819 Department of Pharmacy of Vilnius Medical Society was founded by Vilnius chemists. The main directions of the activity of the members of this department were the following: scientific investigations, solution of practical tasks, spreading of pharmaceutical information, popularization of the knowledge of pharmaceutical science. However, a progressive dynamic development of pharmacy was hindered by unfavorable political and economic factors. In 1917, Lithuanians working in Russia united in a "Society of Lithuanian Pharmacists". The operation of the society was concerned with nationality, which was the main and the only direction of activity. There were four pharmaceutical organizations acting in the Independent Republic of Lithuania (1918-1940). In 1940, after Lithuania's annexation to the USSR, all pharmaceutical professional associations were eliminated. In 1948, Lithuanian Scientific Society of Pharmacists was founded. Despite its professional tasks it had also to proclaim ideological aims. CONCLUSION: Professional activity was influenced by unfavorable political conditions in the country. Independence of social professional associations was limited by czarist Russia, as well as by Soviet authorities. Department of Pharmacy of Vilnius Medical Society had a great influence upon the progress of pharmacy of that time. Associations acting in the Independent Republic of Lithuania were able to defend their business interests, participate in international activity, deal with the affairs of science, the heritage of the pharmacy, and representation of their profession. PMID- 12552168 TI - [Fat embolism]. AB - Fat embolism syndrome remains a rare, but potentially life-threatening complication of long bone fracture and other surgical conditions. The true incidence is difficult to assess as many cases remain undiagnosed. This review aims to discuss the incidence, etiology, mechanical and biochemical theories of pathogenesis, clinical features, diagnosis, treatment, outcome and prophylactic strategies of fat embolism. PMID- 12552169 TI - Facilitating meta-analysis in nursing. PMID- 12552170 TI - Anger in adolescents: sex, ethnicity, age differences, and psychometric properties. AB - BACKGROUND: The State-Trait Anger Expression Inventory (STAXI), a self-report questionnaire, is designed to measure the experience and expression of anger. Reliability and validity of the STAXI have been well established among African and European Americans aged 13 years and older. However, little is known of the use of this instrument among adolescents younger than 13 years and Hispanic American adolescents. OBJECTIVES: Objectives were (a) to test ethnic, sex, and age group differences in STAXI scores in a sample of 11-to-16-year-old African, Hispanic, and European American adolescents; and (b) to assess the psychometric properties of the STAXI among these same adolescents with special emphasis on Hispanic youths, for whom no data are available. METHODS: A cross-sectional design was used with stratified quota sampling techniques. Participants (N = 394) were African, Hispanic, and European Americans aged 11-16 years and were drawn from one public middle school and two public high schools in Houston, Texas. RESULTS: Internal consistency reliability for the anger scales (STAXI) ranged from 0.61 (anger-in) to 0.91 (state-anger) for the younger group (aged 11-13 years), and 0.68 (anger-in) to 0.88 (state-anger) for the older Hispanic Americans (aged 14-16). No notable differences were seen among the three ethnic groups in regards to internal consistency. Results of factor analyses of the five anger scales were similar to those reported originally by the scale author. Ethnicity and age had statistically significant main effects on the anger scales, and there was only one interaction. DISCUSSION: The use of the STAXI among a tri ethnic adolescent population is warranted. The anger-in scale may be less reliable, especially among younger adolescents. PMID- 12552171 TI - Nursing homes as complex adaptive systems: relationship between management practice and resident outcomes. AB - BACKGROUND: Despite numerous clinical and regulatory efforts, problems of poor quality of care in nursing homes continue, suggesting a need for effective management practices. OBJECTIVE: To test complexity hypotheses about the relationship between management practices (communication openness, decision making, relationship-oriented leadership, and formalization) and resident outcomes (aggressive behavior, restraint use, immobility of complications, and fractures), while controlling for case mix, size, ownership, and director's tenure and experience. METHOD: A cross-sectional correlational field study design was used. Primary data were obtained from directors of nursing and registered nurses employed in 164 Texas nursing homes. Investigators administered self report surveys onsite. Secondary data were obtained from 1995 Medicaid Cost Reports and the Texas nursing home Minimum Data Set (MDS) and were linked to primary data using a unique identifier. RESULTS: Hypotheses were supported in that each management practice explained one or more of the resident outcomes. Larger size and longer director of nursing tenure and experience also explained better resident outcomes. Predictors explained 11% n 21% of the variance. DISCUSSION: Complexity science was used to explain the results. The findings open the door to rethinking nursing home management practice. Practices that increase communication and interaction among people are needed for better resident outcomes. PMID- 12552172 TI - Predicting pressure ulcer risk in pediatric patients: the Braden Q Scale. AB - BACKGROUND: While there are valid and reliable pressure ulcer risk assessment tools available for adult patients, none exist for infants and children. To remedy this, the Braden Scale was adapted for use in pediatrics, calling it the Braden Q Scale. OBJECTIVE: The purpose of this study was to: (a) establish the predictive validity of the Braden Q Scale in an acutely ill pediatric population; (b) determine the critical cutoff point for classifying patient risk; and (c) determine the best time to assess patient risk. METHODS: A multisite prospective cohort descriptive study with a convenience sample of 322 patients on bedrest for at least 24 hours without pre-existing pressure ulcers or congenital heart disease were enrolled from three pediatric intensive care units (PICU). The Braden Q score and skin assessment were independently rated and data collectors were blind to the other measures. Patients were observed up to 3 times per week for 2 weeks and then once a week until PICU discharge for a median of 2 observations reflecting 887 skin assessments. RESULTS: Eighty-six patients (27%) developed 199 pressure ulcers; 139 (70%) were Stage I pressure ulcers, 54 (27%) were Stage II pressure ulcers, and 6 (3%) were Stage III pressure ulcers. Most pressure ulcers (57%) were present at the first observation. Using Stage II+ pressure ulcer data obtained during the first observation, a Receiver Operator Characteristic (ROC) curve for each possible score of the Braden Q Scale was constructed. The area under the curve (AUC) was 0.83. At a cutoff score of 16, the sensitivity was 0.88 and the specificity was 0.58. The Braden Q Scale was then modified to eliminate 4 subscales with an AUC <0.7. With 3 subscales (mobility, sensory perception, tissue perfusion/oxygenation) the AUC of this Modified Braden Q Scale was maintained at 0.84. At a cutoff score of 7, the sensitivity was 0.92 and the specificity was 0.59. CONCLUSIONS: The performance of the Braden Q Scale in a pediatric population is similar to that consistently reported for the Braden Scale in adult patients. The Modified Braden Q Scale, with 3 subscales, provides a shorter yet comparable tool. PMID- 12552173 TI - Comparison of two measures of parent-child interaction. AB - BACKGROUND: The numerous parent-child interaction measures available include few that are appropriate to brief clinical encounters or to research settings where time, cost, space, and subject burden are critical factors. OBJECTIVES: This study compares the newer Mother-Infant Communication Screening (MICS) with the established Nursing Child Assessment Satellite Training (NCAST) Teaching Scale (NCAT). METHODS: Theoretical foundations, development, administration, support, and published psychometrics for the two scales are contrasted. Videotapes of 171 caregiver-child interactions in an urban, Hispanic, high-risk sample (children aged 5 months to 36 months) were scored by two trained coders who had established inter-rater reliabilities of.90 (NCAT) and.85 (MICS). RESULTS: Validity correlation coefficients were r =.504, p <.001 for total scores and r =.492, p <.001 for the two most comparable subscales. Distress was defined more narrowly by the MICS. Internal consistencies were.89 to.94 (MICS total scores) (depending on choice of subscales used) and.79 (NCAT). CONCLUSIONS: The NCAT has established strengths for use in clinical practice and research and provides dyadic and separate mother and child scores. The MICS is more feasible for brief clinical screening and performed in this study with promising correlate validity and internal consistency across ages (infancy through 3 years). Interpretation would be strengthened if more clinical studies verifying scores with risk categories were performed and if normative data were established. PMID- 12552174 TI - Black and white men at risk for memory impairment. AB - BACKGROUND: Blacks are under-represented in government-funded research. Epidemiological data indicate that morbidity and mortality in the United States are higher in Blacks than in Whites. OBJECTIVE: To examine memory performance in a sample of Black and White healthy men 70 years of age and older. METHOD: Sixty nine (24 Black, 45 White) men participated in a study of memory performance. All individuals lived either in private homes or subsidized high-rise apartments. RESULTS: Black men had higher anxiety about their memories, used fewer internal and external memory strategies, and had less confidence and less understanding of their everyday memory than White men. Based on their depression scores, more Black men (35%) were at risk for depression than White men (10%). Even though there were no differences in the numbers of men who lived alone, White men were five times more likely to be married and live with a spouse (27 vs. 5). CONCLUSION: In this sample of Black and White men, there were no differences in actual memory performance, but significant differences were found in the subjective aspects of memory evaluation. PMID- 12552175 TI - The relationship between the parturient's positions and perceptions of labor pain intensity. AB - BACKGROUND: While the effect of the maternal position on reducing labor pain has been studied, the data presented to date have not been conclusive. OBJECTIVES: To determine if maternal position reduced the intensity of labor pain during cervical dilatation from 6 to 8 centimeters. METHOD: Pain intensity was measured using the visual analogue scale (VAS) on 39 primiparous and 19 multiparous women (N = 58) who alternately assumed the sitting and supine positions for 15 minutes during cervical dilatation from 6 to 8 centimeters. RESULTS: The pain scores for the sitting position were significantly lower than those for the supine position. The Wilcoxon signed-ranks test showed the VAS scores for the (a) total labor pain ("total" being defined as both abdominal and lumbar pain) during contraction (p =.011), (b) continuous total labor pain (p =.001), (c) lumbar pain during contraction (p <.001), and (d) continuous lumbar pain (p <.001) in the sitting position (significantly lower than in supine position). The diminished pain scores were greater than 13 millimeters, which is the minimum clinically significant change in patient pain severity as measured with the 100 millimeter VAS. The largest decrease occurred in lower back pain. No significant differences were found for abdominal pain scores in either the sitting or supine positions. CONCLUSION: The sitting position offers an effective method to relieve lower back labor pain during cervical dilatation from 6 to 8 centimeters. Similar relief was experienced for women who reported pain only on contraction as well as those with continuous pain. PMID- 12552176 TI - Measuring the frequency of contact between nursing home residents and their family and friends. AB - BACKGROUND: The involvement of family and friends in nursing home care represents an important resource for an overburdened long-term care system. However, little guidance exists for researchers interested in measuring family involvement. OBJECTIVES: This methodological report provides an overview of approaches to measuring family involvement in nursing home care and examines agreement between family and staff on the frequency of visits and telephone calls to a resident by family and friends. Agreement is also assessed for subgroups of the sample based on characteristics of the family, staff, facility, and resident. METHODS: From a large and representative sample of nursing home residents, 823 pairs of significant others and staff were interviewed. Primary variables were reports of visitation and telephone contact received by the resident in the preceding 2 weeks according to the significant other and staff person. RESULTS: Significant other reports of visitation and telephone contact were significantly higher than staff reports (p <.001 and p <.01). Agreement (via intraclass correlation) between significant others and staff was moderate for reports of visit and telephone call frequency. With one exception, no significant differences in agreement were found between subgroups defined by characteristics of the family, staff, facility, or resident. For visits, agreement between nurse's aides and significant others was lower than between other staff persons (e.g., LPNs and RNs) and significant others (p <.05). DISCUSSION: Due to the complexity of nursing home settings as well as of the social support system of residents, researchers need to carefully consider their approach to the measurement of the involvement of family and friends in the nursing home. PMID- 12552177 TI - Assessing publication bias in meta-analysis. AB - BACKGROUND: Although publication bias is a concern when using meta-analysis, techniques for assessing for its potential appear to be underused in nursing. OBJECTIVES: To demonstrate several methods, both graphical and statistical, of assessing for publication bias. METHODS: Four methods are described and demonstrated, ranging from the familiar fail-safe N to the trim-and-fill method, using the same contrived data set. RESULTS: Methods vary in their conclusion about the presence of potential publication bias in the example presented. Some limitations of the methods are noted to assist meta-analysts in determining which to consider using. CONCLUSION: Given that publication bias can affect the validity of meta-analytic studies, researchers are encouraged to use more than one method to assess for bias so that clinicians and decision-makers can have the best possible information available. PMID- 12552178 TI - Using multilevel analysis in patient and organizational outcomes research. AB - BACKGROUND: Outcomes research often compares patient and organizational outcomes across institutions, dealing with variables measured at different hierarchical levels. A traditional approach to analyzing multilevel data has been to aggregate individual-level variables at the institutional level. OBJECTIVES: To introduce the conceptual and statistical background of multilevel analysis and provide an example of multilevel analysis that was used to examine the relationship between nurse staffing and patient outcome. METHODS: A two-level model was presented employing multilevel logistic regression analysis. RESULTS: Outputs from multilevel analysis were interpreted. Other statistics were presented for model specification and testing. CONCLUSION: Researchers should consider multilevel modeling at the study design stage to select theoretically and statistically sound research methods. PMID- 12552179 TI - Smoking cessation. PMID- 12552180 TI - Peer review gives voices to novice researchers. PMID- 12552182 TI - Droperidol: many questions, few answers. PMID- 12552183 TI - Do we need another animal pain model? PMID- 12552184 TI - Antiemetic prophylaxis for office-based surgery: are the 5-HT3 receptor antagonists beneficial? AB - BACKGROUND: Office-based surgery has become increasingly popular because of its cost-saving potential. However, the occurrence of postoperative nausea and vomiting (PONV) can delay patient discharge. Prophylaxis using a combination of antiemetic drugs has been suggested as an effective strategy for minimizing PONV. The authors designed this randomized, double-blinded, placebo-controlled study to assess the efficacy of ondansetron and dolasetron when administered in combination with droperidol and dexamethasone for routine antiemetic prophylaxis against PONV in the office-based surgery setting. METHODS: Following institutional review board approval, 135 consenting outpatients with American Society of Anesthesiologists physical status I-III who were undergoing superficial surgical procedures lasting 20-40 min were randomly assigned to one of three antiemetic treatment groups. Propofol was administered for induction of anesthesia, followed by 2-4% desflurane with 67% nitrous oxide in oxygen. Desflurane was subsequently adjusted to maintain a clinically adequate depth of anesthesia with an electroencephalographic Bispectral Index value between 50 and 60. All patients received 0.625 mg intravenous droperidol and 4 mg intravenous dexamethasone after induction of anesthesia. The study medication, containing normal saline (control), 12.5 mg intravenous dolasetron, or 4 mg intravenous ondansetron, was administered prior to the end of surgery. All patients received local anesthetics at the incisional site and 30 mg intravenous ketolorac to minimize postoperative pain. Recovery profiles, incidence of PONV, requirement for rescue antiemetic drugs, complete response rates, and patient satisfaction were assessed. RESULTS: The recovery times to patient orientation, oral intake, ambulation, and actual discharge did not differ among the three groups. The incidence of PONV, nausea scores, and requirement for rescue antiemetics were also similar in all three groups during the 24-h study period. In addition, the complete response rates to the prophylactic antiemetics (96-98%) and percentages of very satisfied patients (93-98%) were equally high in all three groups. However, the antiemetic drug acquisition costs were US $2.50, $15.50, and $18.50 in the control, dolasetron, and ondansetron groups, respectively. CONCLUSION: The addition of dolasetron (12.5 mg) or ondansetron (4 mg) failed to improve the antiemetic efficacy of droperidol (0.625 mg intravenous) and dexamethasone (4 mg intravenous) when they were used for routine prophylaxis in the office-based surgery setting. PMID- 12552185 TI - Comparison of predicted induction dose with predetermined physiologic characteristics of patients and with pharmacokinetic models incorporating those characteristics as covariates. AB - BACKGROUND: The relationship between patient characteristics and anesthesia induction dose at a high administration rate is unclear. This study was designed to investigate the relation between induction dose and patient characteristics and to compare it to the predicted induction dose using the previously reported pharmacokinetic model. METHODS: Diluted propofol (0.5 mg/ml) dose required to reach loss of consciousness, when infused at an infusion rate per lean body mass (LBM) of 150 mg x kg(-1) x h(-1) (high rate), was determined in 82 patients, ages 10-85 yr. Cardiac output, blood volume, central blood volume (CBV), and hepatic blood flow were measured with indocyanine green pulse spectrophotometry. Stepwise multiple linear regression models were used to investigate the relations between the patient characteristics and induction dose. These were compared with our previously reported parameters at the rate of 40 mg x kg(-1) x h(-1) (low rate) and with predicted induction doses with two previously reported pharmacokinetic models. RESULTS: Significant factors for predicting the induction dose at a high rate were age, LBM, and CBV. Induction dose with one pharmacokinetic model was 1.5 times that of the measured one and the other was half that of the measured one at a high rate. At a low rate, one pharmacokinetic model provided an accurate induction dose. CONCLUSIONS: The prediction of induction dose from physiologic characteristics of patients provides reasonable accuracy at both high and low administration rates of propofol. A previously reported pharmacokinetic model that incorporated patient characteristics provides the same accurate induction dose at a low rate. PMID- 12552186 TI - Effects of isoflurane on gamma-aminobutyric acid type A receptors activated by full and partial agonists. AB - BACKGROUND: Volatile anesthetics prolong inhibitory postsynaptic potentials in central neurons an allosteric action on the gamma-aminobutyric acid type A (GABA(A)) receptor, an effect that may underlie the hypnotic actions of these agents. Inhaled anesthetics such as isoflurane act to enhance responses to submaximal concentrations of GABA, but it is not clear whether their effect is mediated by an increase in the binding of the agonist or by changes in receptor gating behavior. To address this question, the authors studied the effects of isoflurane on a mutant GABA(A) receptor with a gating defect that decreases receptor sensitivity by lowering agonist efficacy. They then compared the effects of clinically relevant concentrations of isoflurane on the actions of GABA and piperidine-4-sulfonic acid (P4S), a partial agonist at the GABA(A) receptor. METHODS: The authors created a mutant of the GABA receptor alpha subunit (L277A) by site-directed mutagenesis. The mutant subunit was coexpressed with beta(2) and gamma(2S) subunits in HEK293 cells, and responses to GABA and P4S were recorded using the whole-cell patch clamp technique. EC values were determined for the full agonist GABA and the partial agonist P4S. The authors also determined the relative efficacy (epsilon) of P4S. These measurements were then repeated in the presence of isoflurane. RESULTS: The concentration-response curve for GABA was shifted to the right (EC(50) = 278 microm) in the alpha(1)(L277A)beta(2)gamma(2S) mutant receptor, compared with the corresponding wild-type alpha(1)beta(2)gamma(2S) GABA(A) receptor (EC(50) = 16 microm). P4S is a partial agonist at both receptors, with a dramatically decreased relative efficacy at the mutant receptor (epsilon = 0.24). When the mutant receptor was studied in the presence of isoflurane, the concentration-response curves for both GABA and P4S were shifted to the left (EC(50) for GABA = 78 microm); the efficacy of P4S also increased significantly (epsilon = 0.40). CONCLUSION: By studying a mutant GABA receptor with impaired gating, the authors were able to demonstrate clearly that isoflurane can increase the efficacy of a partial agonist, as well as increase agonist potency. These data suggest that the volatile anesthetic isoflurane exerts at least some of its effects on the GABA(A) receptor via alterations in gating rather than simply changing binding or unbinding of the agonist. PMID- 12552187 TI - Response surface modeling of remifentanil-propofol interaction on cardiorespiratory control and bispectral index. AB - BACKGROUND: Since propofol and remifentanil are frequently combined for monitored anesthesia care, we examined the influence of the separate and combined administration of these agents on cardiorespiratory control and bispectral index in humans. METHODS: The effect of steady-state concentrations of remifentanil and propofol was assessed in 22 healthy male volunteer subjects. For each subject, measurements were obtained from experiments using remifentanil alone, propofol alone, and remifentanil plus propofol (measured arterial blood concentration range: propofol studies, 0-2.6 microg/ml; remifentanil studies, 0-2.0 ng/ml). Respiratory experiments consisted of ventilatory responses to three to eight increases in end-tidal Pco2 (Petco2). Invasive blood pressure, heart rate, and bispectral index were monitored concurrently. The nature of interaction was assessed by response surface modeling using a population approach with NONMEM. Values are population estimate plus or minus standard error. RESULTS: A total of 94 responses were obtained at various drug combinations. When given separately, remifentanil and propofol depressed cardiorespiratory variables in a dose dependent fashion (resting V(i) : 12.6 +/- 3.3% and 27.7 +/- 3.5% depression at 1 microg/ml propofol and 1 ng/ml remifentanil, respectively; V(i) at fixed Petco of 55 mmHg: 44.3 +/- 3.9% and 57.7 +/- 3.5% depression at 1 microg/ml propofol and 1 ng/ml remifentanil, respectively; blood pressure: 9.9 +/- 1.8% and 3.7 +/- 1.1% depression at 1 microg/ml propofol and 1 ng/ml remifentanil, respectively). When given in combination, their effect on respiration was synergistic (greatest synergy observed for resting V(i)). The effects of both drugs on heart rate and blood pressure were modest, with additive interactions when combined. Over the dose range studied, remifentanil had no effect on bispectral index even when combined with propofol (inert interaction). CONCLUSIONS: These data show dose dependent effects on respiration at relatively low concentrations of propofol and remifentanil. When combined, their effect on respiration is strikingly synergistic, resulting in severe respiratory depression. PMID- 12552188 TI - Effects of EDTA- and sulfite-containing formulations of propofol on respiratory system resistance after tracheal intubation in smokers. AB - BACKGROUND: The formulation of sulfite-containing propofol (SCP) has not been thoroughly investigated in patients with the extensive smoking history for the effects on the total respiratory system resistance after tracheal intubation. However adverse effects, including acute asthma and bronchospasm, have been reported with several other parenteral formulations of drugs containing sulfite as preservative. Therefore, the aim of this prospective randomized and double blind study was to investigate the effects of EDTA-containing propofol (ECP) and SCP on total respiratory system resistance (Rrs) in patients with the prolonged smoking history and undergoing propofol-based total intravenous anesthesia with tracheal intubation. METHODS: 40 patients scheduled for general anesthesia were enrolled into the study. Anesthesia was induced with either 2 mg/kg ECP, or 2 mg/kg SCP followed by vecuronium (0.1 mg/kg) to ensure complete neuromuscular relaxation for the time of the study. Maintenance anesthesia was continued with propofol infusion at 0.15 mg/kg/min for the first 15 min after intubation. Total respiratory system resistance (Rrs), was measured continuously for 10 min postintubation. RESULTS: The analysis of repeated Rrs measurements taken every minute for 10 min postintubation revealed trend consisting of higher Rrs in the SCP group when compared to the ECP group. The statistical analysis of the data performed using repeated measures analysis of covariance demonstrated statistically significant effect (P < 0.05) of the treatment group factor (SCP vs. ECP) and the time factor (time after intubation) on the postintubation Rrs. CONCLUSION: The total respiratory system resistance measured repeatedly for 10 min after tracheal intubation in patients with smoking history is significantly elevated after induction with SCP than after induction with ECP. The preservative used for propofol formulation may alter the effects of propofol on the total respiratory system resistance in smokers. PMID- 12552189 TI - Intracranial pressure and cerebral hemodynamic in patients with cerebral tumors: a randomized prospective study of patients subjected to craniotomy in propofol fentanyl, isoflurane-fentanyl, or sevoflurane-fentanyl anesthesia. AB - BACKGROUND: A critical point during craniotomy is opening of dura, where a high intracranial pressure (ICP) results in swelling of cerebral tissue. Controlled studies concerning ICP, degree of dural tension, and degree of cerebral swelling are therefore warranted. METHODS: In an open-label study, 117 patients with supratentorial cerebral tumors were randomized to propofol-fentanyl (group 1), isoflurane-fentanyl (group 2), or sevoflurane-fentanyl anesthesia (group 3). Normo- to moderate hypocapnia was applied, with a target level of arterial carbon dioxid tension of 30-40 mmHg. Mean arterial blood pressure was stabilized with intravenous ephedrine (2.5-5 mg) if necessary. Subdural ICP, mean arterial blood pressure, cerebral perfusion pressure (CPP), arteriovenous oxygen difference (AVDo2), internal jugular vein oxygen saturation were monitored before and after a 10-min period of hyperventilation, and the carbon dioxide reactivity was calculated. Furthermore, the tension of dura before and during hyperventilation and the degree of cerebral swelling during hyperventilation and after opening of the dura were estimated by the neurosurgeon. RESULTS: No differences were found between groups with regard to demographics, neuroradiologic examination, positioning of the head, and time to ICP measurement. Before and during hyperventilation, ICP was significantly lower and mean arterial blood pressure and CPP significantly higher in group 1 compared with groups 2 and 3 (P < 0.05). The tension of dura before and during hyperventilation was significantly lower in group 1 compared with group2 (P < 0.05), but not significantly different from group 3. In group 1, cerebral swelling after opening of dura was significantly lower compared with groups 2 and 3 (P < 0.05). Furthermore, AVDo was significantly higher and jugular vein oxygen saturation and carbon dioxide reactivity were significantly lower in group 1 compared with groups 2 and 3 (P < 0.05). No significant differences with regard to ICP, CPP, AVDo, carbon dioxide reactivity, and jugular vein oxygen saturation were found between patients anesthetized with isoflurane and sevoflurane. CONCLUSIONS: The study indicates that before as well as during hyperventilation, subdural ICP and AVDo2 are lower and CPP higher in propofol-anesthetized patients compared with patients anesthetized with isoflurane or sevoflurane. These findings were associated with less tendency for cerebral swelling after opening of dura in the propofol group. The carbon dioxide reactivity in patients anesthetized with isoflurane and sevoflurane was significantly higher than in the propofol group. The differences in subdural ICP between the groups are presumed to be caused by differences in the degree of vasoconstriction elicited by the anesthetic agents, but autoregulatory mechanisms caused by differences in CPP cannot be excluded. PMID- 12552190 TI - Antifibrinolytic therapy and perioperative blood loss in cancer patients undergoing major orthopedic surgery. AB - BACKGROUND: Aprotinin has been reported to reduce blood loss and transfusion requirements in patients having major orthopedic operations. Data on whether epsilon amino-caproic acid (EACA) is effective in this population are sparse. METHODS: Sixty-nine adults with malignancy scheduled for either pelvic, extremity or spine surgery during general anesthesia entered this randomized, double-blind, placebo-controlled trial, and received either intravenous aprotinin (n = 23), bolus of 2 x 10(6) kallikrein inactivator units (KIU), followed by an infusion of 5 x 10(5) KIU/h, or EACA (n = 22), bolus of 150 mg/kg, followed by a 15 mg/kg/h infusion or saline placebo (n = 24) during surgery. Our goal was to determine whether prophylactic EACA or aprotinin therapy would reduce perioperative blood loss (intraoperative + first 48h) >30% when compared to placebo. RESULTS: The mean age of the study population was 52 +/- 17 yr. The groups did not differ in age, duration of surgery, perioperative blood loss or number of packed erythrocyte units transfused. When compared to the placebo group, the two treated groups had a significantly lower D-Dimer level immediately after surgery, P < 0.01. CONCLUSIONS: Under the conditions of this study, we were unable to find a clinical benefit to using aprotinin or EACA to reduce perioperative blood loss or transfusion requirements during major orthopedic surgery in cancer patients. PMID- 12552191 TI - Cardiopulmonary bypass decreases G protein-coupled receptor kinase activity and expression in human peripheral blood mononuclear cells. AB - BACKGROUND: Cardiopulmonary bypass (CPB) has been implicated in the development of organ injury associated with cardiac surgery. At the molecular level, CPB is accompanied by a pronounced proinflammatory response including an increase in plasma interleukin (IL)-6. The IL-6 has been shown to be increased in rheumatoid arthritis, a chronic inflammatory disease, where it has been implicated in decreasing G protein-coupled receptor kinases (GRKs) in peripheral blood mononuclear cells. Since IL-6 is substantially increased after CPB, the study tested whether the increase of IL-6 during CPB leads to a decrease of GRKs in mononuclear cells. This is important because GRKs regulate the function of G protein-coupled receptors involved in inflammation. METHODS: Fifteen patients had blood withdrawn before CPB, 2 h after CPB, and on postoperative day one (POD1). Plasma IL-6 concentrations were determined by enzyme-linked immunosorbent assay. The GRK protein expression and activity were determined by Western blot and phosphorylation of rhodopsin using [gamma-(32)P] adenosine triphosphate, respectively. RESULTS: Plasma IL-6 increased over 20-fold after CPB and remained increased on POD1. Cytosolic GRK activity in mononuclear cells decreased by 39 +/ 29%; cytosolic GRK2 and membrane-bound GRK6 decreased by 90 +/- 15 and 65 +/- 43%, respectively. The GRK activity and expression of GRK2/GRK6 on POD1 returned to basal levels in many but not all patients. CONCLUSIONS: The CPB causes a profound decrease in mononuclear cell GRKs, and the recovery of these kinases on POD1 is quite variable. The significance of the variable recovery of GRKs after CPB and their potential role as a marker of clinical outcome deserves further investigation. PMID- 12552192 TI - What is the minimum training required for successful cricothyroidotomy?: a study in mannequins. AB - BACKGROUND: A correctly performed cricothyroidotomy may be lifesaving in a cannot ventilate, cannot-intubate situation. However, many practicing anesthesiologists do not have experience with cricothyroidotomy. The purpose of this study was to determine the minimum training required to perform cricothyroidotomy in 40 s or less in mannequins. METHODS: After informed consent, participants were shown a demonstration video and asked to perform 10 consecutive cricothyroidotomy procedures on a mannequin using a preassembled percutaneous dilational cricothyroidotomy set. Each attempt was timed from skin palpation to lung insufflation. Cricothyroidotomy was considered successful if it was performed in 40 s or less, and the cricothyroidotomy time was considered to have plateaued when there were no significant reductions in cricothyroidotomy times in three consecutive attempts. RESULTS: One hundred two anesthesiologists participated in the study. There was a significant reduction of cricothyroidotomy times over the 10 attempts (P < 0.0001) and between three consecutive attempts until the fourth attempt (P < 0.03). The cricothyroidotomy times plateaued by the fourth attempt, while the success rate plateaued at the fifth attempt (94, 96, 96, and 96% at the fourth, fifth, sixth, and seventh attempts, respectively). CONCLUSION: Practice on mannequins leads to reductions in cricothyroidotomy times and improvement in success rates. By the fifth attempt, 96% of participants were able to successfully perform the cricothyroidotomy in 40 s or less. While clinical correlates are not known, the authors recommend that providers of emergency airway management be trained on mannequins for at least five attempts or until their cricothyroidotomy time is 40 s or less. The most appropriate retraining intervals have yet to be determined for optimal cricothyroidotomy skill retention. PMID- 12552193 TI - The Parker Flex-Tip tube versus a standard tube for fiberoptic orotracheal intubation: a randomized double-blind study. AB - BACKGROUND: During fiberoptic tracheal intubation, passage of the fiberscope itself to the trachea is often fairly easy, but passage of the tube into the trachea may be difficult or even impossible. A new type of disposable endotracheal tube, the Parker Flex-Tip tube, has a tip that reduces the gap between the fiberscope and the inside of the tube. Thus, theoretically, a smaller risk of impinging on laryngeal structures during insertion in trachea is expected. METHODS: Eighty patients scheduled for elective anesthesia using orotracheal intubation were randomized to either a Parker Flex-Tip tube or a standard (Portex) 7.5-mm-ID endotracheal tube. Blinding was obtained by having the tube premounted on the fiberscope (Olympus LF-1; diameter of fiberscope = 4 mm) and thereafter covered with a black opaque plastic bag. Difficulty in placing the tube was scored using an objective standardized grading system. RESULTS: Seventy-six patients completed the study. The use of the Parker Flex-Tip tube reduced the incidence of need for repositioning of the tube during insertion into trachea from 89% to 29% (P < 0.0001) when compared to the standard tube. The median time for passage of the tube into the trachea was reduced from 20 s to 7.5 s (P < 0.0001). CONCLUSIONS: During oral fiberoptic intubation, the use of the Parker Flex-Tip tube is associated with greater incidence of initial success of passage of the tube into trachea when compared to a standard endotracheal tube. PMID- 12552194 TI - Phonomyography as a novel method to determine neuromuscular blockade at the laryngeal adductor muscles: comparison with the cuff pressure method. AB - BACKGROUND: Neuromuscular blockade at the laryngeal adductor muscles may be measured using the cuff of a endotracheal tube placed between the vocal cords. Phonomyography is an alternative method of neuromuscular monitoring. In this study, phonomyography is applied to determine blockade at the larynx and compared with the cuff pressure method. METHODS: After the authors obtained approval from the ethics committee and informed consent, 28 patients were entered in the study. After induction of anesthesia, a endotracheal tube was inserted. Its cuff was placed in the trachea in routine fashion (n = 14) or between the vocal cords (n = 14). In all patients, a small condenser microphone was placed in the vestibular fold, just lateral to the tube, next to the laryngeal adductor muscles. The recurrent laryngeal nerve was stimulated supramaximally with single twitch stimulation (0.1 Hz) for onset, and train-of-four stimulation every 12 s during offset of neuromuscular blockade was produced by 0.1 mg/kg mivacurium. Onset and recovery of neuromuscular blockade measured by the two methods were compared using the test (P < 0.05), and a Bland-Altman test was performed to define agreement between the two methods. Onset and recovery of neuromuscular blockade measured by phonomyography with the cuff placed between the vocal cords or in the trachea were compared using the test (P < 0.05). RESULTS: Mean onset, maximum effect, and time to reach 25% and 75% of control twitch response for phonomyography cuff pressure method were 145 s (SD, 25) 156 s (SD, 33), 89% (SD, 4) 91% (SD, 4), 9 min (SD, 4) 10 min (SD, 3), and 27 min (SD, 4) 29 min (SD, 4), respectively, without being significantly different. Mean bias was -2%, with limits of agreement of -20 and +18% for all signals (cuff method minus phonomyography). There was no significant difference in onset and offset of neuromuscular blockade measured using phonomyography with the cuff placed between the vocal cords or in the trachea. CONCLUSIONS: Both methods can be used interchangeably to determine neuromuscular blockade of the laryngeal adductor muscles. Phonomyography allows measurement of laryngeal blockade with the endotracheal tube in the normal position. PMID- 12552195 TI - Effects of isoflurane and propofol on glutamate and GABA transporters in isolated cortical nerve terminals. AB - BACKGROUND: Depression of glutamate-mediated excitatory transmission and potentiation of gamma-aminobutyric acid (GABA)-mediated inhibitory transmission appear to be primary mechanisms by which general anesthetics produce anesthesia. Since effects on transmitter transport have been implicated in anesthetic actions, the authors examined the sensitivity of presynaptic glutamate and GABA transporters to the effects of a representative volatile (isoflurane) and a representative intravenous (propofol) anesthetic. METHODS: A dual-isotope (l [3H]glutamate and [14C]GABA) approach allowed simultaneous comparisons of anesthetic effects on three independent assays of glutamate and GABA transporters in adult rat cerebral cortex: transmitter uptake into isolated nerve terminals (synaptosomes), transmitter binding to lysed and washed synaptosomes (synaptic membranes), and carrier-mediated release (reverse transport) of transmitter from preloaded synaptosomes using a modified superfusion system. RESULTS: Isoflurane produced small but statistically significant inhibition of l-[3H]glutamate and [14C]GABA uptake, while propofol had no effect. Inhibition of uptake by isoflurane was noncompetitive, an outcome that was mimicked by indirectly affecting transporter function through synaptosomal depolarization. Neither isoflurane nor propofol affected l-[3H]glutamate or [14C]GABA binding to synaptic membranes or Ca(2+)-independent carrier-mediated l-[3H]glutamate or [14C]GABA release (reverse transport). CONCLUSIONS: These findings suggest that isoflurane and propofol at clinical concentrations do not affect excitatory glutamatergic transmission or inhibitory GABAergic transmission directly effects on their presynaptic neuronal transporters. PMID- 12552196 TI - Hepatic ischemia is associated with an increase in liver parenchyma nitric oxide that is in part enzyme-independent. AB - BACKGROUND: Nitric oxide (NO) might be involved in liver response to local ischemia-reperfusion injury. METHODS: A specific NO-sensitive electrode was inserted into liver parenchyma of anesthetized rabbits. After a 45-min period of stable NO signal, the vascular pedicle of the caudal lobe of the liver was clamped for 45 min, then the clamp was removed. Perfusion of the right upper lobe was left unchanged. The same procedure was applied in other animals after administration of a long-acting nonspecific NO synthase inhibitor NAPNA. RESULTS: Occlusion of the caudal pedicle was associated with a mean threefold increase in NO signal measured in the caudal lobe. After unclamping, this signal returned within 8 min to baseline value and remained stable for the next 6 h. In the right upper lobe, NO signal was unaffected by caudal lobe ischemia. By the end of the 6 h reperfusion period, administration of the NO inhibitor l-NAME led to a suppression of the NO signal, thus demonstrating the specificity of the measurement. Plasma nitrate and nitrite concentrations remained almost unchanged during the study period in all groups. In animals whose NO synthases had been previously inhibited by NAPNA, clamping the caudal pedicle for 45 min was still associated with a significant increase in caudal lobe NO signal. CONCLUSION: Nitric oxide is present in liver parenchyma, and its generation is dramatically affected by an ischemia injury. The increased NO generation during local ischemia is, at least in part, independent of NO synthases. PMID- 12552197 TI - Isoflurane alters energy substrate metabolism to preserve mechanical function in isolated rat hearts following prolonged no-flow hypothermic storage. AB - BACKGROUND: Isoflurane enhances mechanical function in hearts subject to normothermic global or regional ischemia. The authors examined the effectiveness of isoflurane in preserving mechanical function in hearts subjected to cardioplegic arrest and prolonged hypothermic no-flow storage. The role of isoflurane in altering myocardial glucose metabolism during storage and reperfusion during these conditions and the contribution of adenosine triphosphate-sensitive potassium (K(atp)) channel activation in mediating the functional and metabolic effects of isoflurane preconditioning was determined. METHODS: Isolated working rat hearts were subjected to cardioplegic arrest with St. Thomas' II solution, hypothermic no-flow storage for 8 h, and subsequent aerobic reperfusion. The consequences of isoflurane treatment were assessed during the following conditions: (1) isoflurane exposure before and during storage; (2) brief isoflurane exposure during early nonworking poststorage reperfusion; and (3) isoflurane preconditioning before storage. The selective mitochondrial and sarcolemmal K(atp) channel antagonists, 5-hydroxydecanoate and HMR 1098, respectively, were used to assess the role of K(atp) channel activation on glycogen consumption during storage in isoflurane-preconditioned hearts. RESULTS: Isoflurane enhanced recovery of mechanical function if present before and during storage. Isoflurane preconditioning was also protective. Isoflurane reduced glycogen consumption during storage under the aforementioned circumstances. Storage of isoflurane-preconditioned hearts in the presence of 5 hydroxydecanoate prevented the reduction in glycogen consumption during storage and abolished the beneficial effect of isoflurane preconditioning on recovery of mechanical function. CONCLUSIONS: Isoflurane provides additive protection of hearts subject to cardioplegic arrest and prolonged hypothermic no-flow storage and favorably alters energy substrate metabolism by modulating glycolysis during ischemia. The effects of isoflurane preconditioning on glycolysis during hypothermic no-flow storage appears to be associated with activation of mitochondrial K(atp) channels. PMID- 12552198 TI - Preconditioning with sevoflurane reduces changes in nicotinamide adenine dinucleotide during ischemia-reperfusion in isolated hearts: reversal by 5 hydroxydecanoic acid. AB - BACKGROUND: Ischemia causes an imbalance in mitochondrial metabolism and accumulation of nicotinamide adenine dinucleotide (NADH). We showed that anesthetic preconditioning (APC), like ischemic preconditioning, improved mitochondrial NADH energy balance during ischemia and improved function and reduced infarct size on reperfusion. Opening adenosine triphosphate-sensitive potassium (K(atp)) channels may be involved in triggering APC. The authors tested if effects of APC on NADH concentrations before, during, and after ischemia are reversible by 5-hydroxydecanoate (5-HD), a putative mitochondrial K channel blocker. METHODS: Nicotinamide adenine dinucleotide fluorescence was measured in 60 guinea pig Langendorff-prepared hearts assigned into five groups: (1) no treatment before ischemia; (2) APC by exposure to 1.3 mm sevoflurane for 15 min; (3) 200 microm 5-HD from 5 min before to 15 min after sevoflurane exposure; (4) 35 min 5-HD alone; and (5) no treatment and no ischemia. Sevoflurane was washed out for 30 min, and 5-HD for 15 min, before 30-min ischemia and 120-min reperfusion. RESULTS: Nicotinamide adenine dinucleotide was reversibly increased during sevoflurane exposure before ischemia, and the increase and rate of decline in NADH during ischemia were reduced after APC. 5-HD abolished these changes in NADH. On reperfusion, function was improved and infarct size reduced after APC compared with other groups. CONCLUSION: Anesthetic preconditioning was evidenced by improved mitochondrial bioenergetics as assessed from NADH concentrations during ischemia and by attenuated reperfusion injury. Reversal of APC by bracketing sevoflurane exposure with 5-HD suggests that APC is triggered by mitochondrial K channel opening or, alternatively, by attenuated mitochondrial respiration without direct involvement of mitochondrial K channel opening. PMID- 12552199 TI - Isoflurane decreases ATP sensitivity of guinea pig cardiac sarcolemmal KATP channel at reduced intracellular pH. AB - BACKGROUND: Volatile anesthetics can protect the myocardium against ischemic injury by opening the adenosine triphosphate (ATP)-sensitive potassium (K(atp)) channels. However, direct evidence for anesthetic-channel interaction is still limited, and little is known about the role K(atp) channel modulators play in this effect. Because pH is one of the regulators of K(atp) channels, the authors tested the hypothesis that intracellular pH (pHi) modulates the direct interaction of isoflurane with the cardiac K(atp) channel. METHODS: The effects of isoflurane on sarcolemmal K(atp) channels were investigated at pHi 7.4 and pHi 6.8 in excised inside-out membrane patches from ventricular myocytes of guinea pig hearts. RESULTS: At pHi 7.4, intracellular ATP (1-1,000 microm) inhibited K(atp) channels and decreased channel open probability (Po) in a concentration dependent manner with an IC(50) of 8 +/- 1.5 microm, and isoflurane (0.5 mm) either had no effect or decreased channel activity. Lowering pHi from 7.4 to 6.8 enhanced channel opening by increasing Po and reduced channel sensitivity to ATP, with IC shifting from 8 +/- 1.2 to 45 +/- 5.6 microm. When applied to the channels activated at pHi 6.8, isoflurane (0.5 mm) increased Po and further reduced ATP sensitivity, shifting IC(50) to 110 +/- 10.0 microm. CONCLUSIONS: Changes in pHi appear to modulate isoflurane interaction with the cardiac K(atp) channel. At pHi 6.8, which itself facilitates channel opening, isoflurane enhances channel activity by increasing Po and reduces sensitivity to inhibition by ATP without changing the unitary amplitude of single channel current or the conductance. These results support the hypothesis of direct isoflurane-K(atp) channel interaction that may play a role in cardioprotection by volatile anesthetics. PMID- 12552200 TI - Interaction of isoflurane with the dopamine transporter. AB - BACKGROUND: Isoflurane administration is known to increase extracellular dopamine (DA) concentration. Because the dopamine transporter (DAT) is a key regulator of DA, it is likely affected by isoflurane. This study investigates the hypothesis that isoflurane inhibits DA reuptake by causing DAT to be trafficked into the cell. METHODS: Rhesus monkeys were scanned with positron emission tomography (PET) using [18F]FECNT (a highly specific DAT ligand) while anesthetized with 1% isoflurane. The isoflurane was increased to 2%, and the animals were rescanned. Uptake was analyzed with the tissue reference method using the cerebellum as the reference tissue to determine the binding potential in the putamen. Immunohistochemistry and Western blot analyses were performed in rats to determine if isoflurane administration would change the total amount of DAT. Rats breathed air plus 2% isoflurane for 30 min, and then striatal DAT assays were rapidly performed. immunocytochemistry experiments were performed using human embryonic kidney (HEK) cells stably transfected with human DAT. The cells were exposed to 4% isoflurane for 1 h while the location of DAT was observed with fluorescent confocal microscopy. RESULTS: The [18F]FECNT binding potential in rhesus monkeys decreased by 63 +/- 6% (SEM, n = 5) when isoflurane was increased from 1 to 2% as compared with no significant change (0.7 +/- 2.5%; SEM, n = 5) when the isoflurane concentration was not changed (P < 0.001). No difference in DAT staining between isoflurane-treated and control rats was apparent from visual inspection, and quantitative Western blot analyses showed no significant change in total DAT protein. After isoflurane treatment, focal puncta of intense fluorescence was visible inside the HEK cells. CONCLUSIONS: The experiments indicate that DAT is trafficked into the cell by isoflurane without changing the total amount of DAT in the striatum. The PET data are consistent with this finding, provided that intracellular DAT acquires a conformation that has low affinity for [18F]FECNT. Thus, [18F]FECNT appears to be an excellent agent for measuring plasma membrane-expressed DAT and evaluating DAT trafficking. PMID- 12552201 TI - Despite in vitro increase in cyclic guanosine monophosphate concentrations, intracarotid nitroprusside fails to augment cerebral blood flow of healthy baboons. AB - BACKGROUND: During cerebral angiography, intracarotid infusion of sodium nitroprusside (SNP), an endothelium-independent nitric oxide donor, fails to increase cerebral blood flow (CBF) of human subjects. A confounding effect of intracranial pathology or that of radiocontrast could not be ruled out in these experiments. The authors hypothesized that, if nitric oxide was a significant regulator of CBF of primates, then intracarotid SNP will augment CBF of baboons. METHODS: In studies, CBF (intraarterial (133)Xe technique) was measured in healthy baboons during isoflurane anesthesia at (1) baseline and during (2) induced hypertension with intravenous phenylephrine, (3) concurrent infusions of intravenous phenylephrine and intracarotid SNP, and (4) intracarotid verapamil (positive control drug). In studies, the authors measured tissue cyclic guanosine monophosphate (cGMP) by radioimmunoassay after incubating vascular rings obtained from freshly killed baboons (1) with increasing concentrations of SNP and (2) after SNP exposure following preincubation with the radiocontrast agent, iohexhol. RESULTS: In the studies, coinfusion of intravenous phenylephrine and intracarotid SNP did not increase CBF. However, intracarotid verapamil significantly increased CBF (from 26 +/- 7 to 43 +/- 11 ml x 100 g(-1) x min(-1); P < 0.0001) without a change in mean arterial pressure. In the studies, incubation of intracranial arterial rings in SNP resulted in dose-dependent increases in cGMP concentrations. A similar increase in cGMP content was evident despite iohexhol preincubation. CONCLUSIONS: Collectively, these results suggest that, in healthy baboons, intracarotid SNP does not decrease arteriolar resistance, although SNP could affect proximal arterial tone, as demonstrated by the increase in cGMP content of these vessels. PMID- 12552202 TI - Respiratory depression by tramadol in the cat: involvement of opioid receptors. AB - BACKGROUND: Tramadol hydrochloride (tramadol) is a synthetic opioid analgesic with a relatively weak affinity at opioid receptors. At analgesic doses, tramadol seems to cause little or no respiratory depression in humans, although there are some conflicting data. The aim of this study was to examine whether tramadol causes dose-dependent inhibitory effects on the ventilatory carbon dioxide response curve and whether these are reversible or can be prevented by naloxone. METHODS: Experiments were performed in cats under alpha-chloralose-urethane anesthesia. The effects of tramadol and naloxone were studied by applying square wave changes in end-tidal pressure of carbon dioxide (Petco2; 7.5-11 mmHg) and by analyzing the dynamic ventilatory responses using a two-compartment model with a fast peripheral and a slow central component, characterized by a time constant, carbon dioxide sensitivity, time delay, and a single offset (apneic threshold). RESULTS: In five animals 1, 2, and 4 mg/kg tramadol (intravenous) increased the apneic threshold (control: 28.3 +/- 4.8 mmHg [mean +/- SD]; after 4 mg/kg: 36.7 +/- 7.1 mmHg; P < 0.05) and decreased the total carbon dioxide sensitivity (control: 109.3 +/- 41.3 ml x min(-1) x mmHg(-1) ) by 31, 59, and 68%, respectively, caused by proportional equal reductions in sensitivities of the peripheral and central chemoreflex loops. Naloxone (0.1 mg/kg, intravenous) completely reversed these effects. In five other cats, 4 mg/kg tramadol caused an approximately 70% ventilatory depression at a fixed Pet co2 of 45 mmHg that was already achieved after 15 min. A third group of five animals received the same dose of tramadol after pretreatment with naloxone. At a fixed Petco of 45 mmHg, naloxone prevented more than 50% of the expected ventilatory depression in these animals. CONCLUSIONS: Because naloxone completely reversed the inhibiting effects of tramadol on ventilatory control and it prevented more than 50% of the respiratory depression after a single dose of tramadol, the authors conclude that this analgesic causes respiratory depression that is mainly mediated by opioid receptors. PMID- 12552203 TI - The alpha2-adrenoceptor agonist dexmedetomidine converges on an endogenous sleep promoting pathway to exert its sedative effects. AB - BACKGROUND: The authors investigated whether the sedative, or hypnotic, action of the general anesthetic dexmedetomidine (a selective alpha -adrenoceptor agonist) activates endogenous nonrapid eye movement (NREM) sleep-promoting pathways. METHODS: c-Fos expression in sleep-promoting brain nuclei was assessed in rats using immunohistochemistry and hybridization. Next, the authors perturbed these pathways using (1) discrete lesions induced by ibotenic acid, (2) local and systemic administration of gamma-aminobutyric acid receptor type A (GABA ) receptor antagonist gabazine, or (3) alpha2-adrenoceptor antagonist atipamezole in rats, and (4) genetic mutation of the alpha -adrenoceptor in mice. RESULTS: Dexmedetomidine induced a qualitatively similar pattern of c-Fos expression in rats as seen during normal NREM sleep, a decrease in the locus ceruleus (LC) and tuberomammillary nucleus (TMN) and an increase in the ventrolateral preoptic nucleus (VLPO). These changes were attenuated by atipamezole and were not seen in mice lacking functional alpha2a-adrenoceptors, which do not show a sedative response to dexmedetomidine. Bilateral VLPO lesions attenuated the sedative response to dexmedetomidine, and the dose-response curve to dexmedetomidine was shifted right by gabazine administered systemically or directly into the TMN. VLPO lesions and gabazine pretreatment altered c-Fos expression in the TMN but in not the LC after dexmedetomidine administration, indicating a hierarchical sequence of changes. CONCLUSIONS: The authors propose that endogenous sleep pathways are causally involved in dexmedetomidine-induced sedation; dexmedetomidine's sedative mechanism involves inhibition of the LC, which disinhibits VLPO firing. The increased release of GABA at the terminals of the VLPO inhibits TMN firing, which is required for the sedative response. PMID- 12552204 TI - Thermogenesis inhibition in brown adipocytes is a specific property of volatile anesthetics. AB - BACKGROUND: This investigation examined the possibility that the inhibitory effect of halothane on nonshivering thermogenesis (heat production) in brown adipocytes is not a universal effect of all anesthetic agents but related to the type of anesthetic. METHODS: Brown adipocytes from hamster were isolated with a collagenase digestion method and incubated with anesthetic agents. The rate of oxygen consumption was measured with an oxygen electrode. The effect of clinically relevant (and higher) doses of anesthetics of different classes on basal and norepinephrine-induced thermogenesis (oxygen consumption) was tested. RESULTS: Two distinct groups of anesthetics could be distinguished: thermogenesis inhibitors and noninhibitors. Thermogenesis inhibitors include volatile anesthetics such as halothane (IC(50), 1.1 mm), ether (IC(50), 20 mm), and chloroform (IC(50), 2.2 mm) (nominal concentrations), but also tribromoethanol (IC(50), 0.6 mm), all inducing inhibition of norepinephrine-induced thermogenesis without affecting the EC for norepinephrine. Thermogenesis noninhibitors include the nonvolatile anesthetics pentobarbital, propofol, ketamine, and urethane, the inhalation anesthetic nitrous oxide, and, notably, also the volatile nonanesthetics (nonimmobilizers) 1,2-dichlorohexafluorocyclobutane and 2,3 dichlorooctafluorobutane; none of these compounds had any effect on norepinephrine-induced thermogenesis at any concentration tested. CONCLUSIONS: There are two distinct classes of anesthetics with regard to effects on thermogenesis, thermogenesis inhibitors and thermogenesis noninhibitors. The results are important for the interpretation of studies in thermal biology in general; specifically, they indicate that conclusions concerning regulation of nonshivering thermogenesis during anesthesia depend on the type of anesthetic used. Of clinical importance is that the volatile anesthetics are inhibitory for nonshivering thermogenesis and thus for an alternative heat production when myorelaxants prevent shivering. As the distinction between thermogenesis inhibitors and thermogenesis noninhibitors corresponds to the distinction between volatile and nonvolatile anesthetics, it may be related to the mode of action of the volatile anesthetics. PMID- 12552205 TI - Ketamine and midazolam differentially inhibit nonadrenergic noncholinergic lower esophageal sphincter relaxation in rabbits: role of superoxide anion and nitric oxide synthase. AB - BACKGROUND: The authors previously reported that ketamine and midazolam inhibited nitric oxide-mediated nonadrenergic noncholinergic (NANC) lower esophageal sphincter (LES) relaxation nitric oxide-3',5'-cyclic guanosine monophosphate pathway modulation. The mechanisms inhibiting the NANC relaxation by ketamine and midazolam were investigated. METHODS: The isometric tension of circular distal esophageal muscle strips from Japanese White rabbits was examined. NANC relaxation was induced by KCl (30 mm) in the presence of atropine (3 x 10(-6) m) and guanethidine (3 x 10(-6) m). Nitric oxide synthase activity in the absence and presence of ketamine and midazolam was analyzed using the biochemical conversion of L-[3H]arginine to L-[3H]citrulline. RESULTS: The ketamine-induced inhibition of the NANC relaxation was partly reversed by superoxide dismutase (200, 400 U/ml) but not by catalase (100 U/ml). Ketamine concentration dependently inhibited the relaxation induced by N-ethylethanamine:1,1-diethyl-2 hydroxy-2-nitrosohydrazine (diethylamine NONOate) and S-nitrosoglutathione. The NANC relaxation itself was not affected by superoxide dismutase. The midazolam induced inhibition of the NANC relaxation was reversed neither by superoxide dismutase nor by catalase, and midazolam did not affect the relaxations induced by nitric oxide donors. The nitric oxide synthase activity was concentration dependently suppressed by midazolam, but there was no marked effect of ketamine. Pyrogallol, a superoxide generator, inhibited the NANC and the diethylamine NONOate-induced relaxations. The pyrogallol-induced inhibition of the NANC relaxation was reversed by superoxide dismutase. CONCLUSION: These findings suggest that ketamine inhibits NANC LES relaxation by the extracellular production of superoxide anion, and that midazolam inhibits it by the inhibition of nitric oxide synthase activity. PMID- 12552206 TI - Supraspinal antinociceptive effects of mu and delta agonists involve modulation of adenosine uptake. AB - BACKGROUND: The modulation of extracellular adenosine concentration by opioids provides evidence that the antinociceptive effects of these compounds involve endogenous adenosine. The aim of this study was to determine whether there is a relation between the inhibition of brain synaptosome adenosine uptake by opioid agonists and the analgesic effects of these compounds. METHODS: The authors used the hot plate and tail-pinch tests to evaluate in mice (C57BL/6 females; weight, 25-30 g) the effects of caffeine, a nonspecific adenosine receptor antagonist, on the antinociceptive effect induced by the intracerebroventricular administration of oxymorphone as a mu agonist, SNC80 as a delta agonist, or U69593 as a kappa agonist. They also investigated the effect of these opioid receptor agonists on the uptake of adenosine by whole brain synaptosomes. RESULTS: Caffeine decreased the analgesic effects induced by oxymorphone or SNC80 but not those induced by U69593. Oxymorphone and SNC80 inhibited adenosine uptake by brain cells, but U69593 did not. CONCLUSION: The antinociceptive effects obtained with mu or delta (but not kappa) agonists administered supraspinally were indicative of the involvement of modulation of adenosine uptake. PMID- 12552207 TI - Neuroprotective effect of urinary trypsin inhibitor against focal cerebral ischemia-reperfusion injury in rats. AB - BACKGROUND: Acute inflammatory reactions cause neuronal damage in cerebral ischemia-reperfusion. Urinary trypsin inhibitor (UTI), a serine protease inhibitor, is cytoprotective against ischemia-reperfusion injury in the liver, intestine, kidney, heart, and lung through its antiinflammatory activity. Neuroprotective action of UTI on transient global cerebral ischemia has been documented. This is the first study to determine whether UTI is neuroprotective against transient focal cerebral ischemia. METHODS: Adult male Wistar rats were randomly assigned to the following treatment groups: 0.9% saline (control, n = 9); 100,000 U/kg UTI (n = 9); and 300,000 U/kg UTI (n = 9). Treatments were performed intravenously 10 min before right middle cerebral artery occlusion for 2 h and subsequent reperfusion. Ninety-six hours after the onset of reperfusion, the motor neurologic deficit and the cerebral infarct size were evaluated. Furthermore, immunohistochemical staining for myeloperoxidase and nitrotyrosine to count infiltrating neutrophils and nitrated cells, respectively, was performed on the brain sections. RESULTS: Infarct volume in the 300,000 U/kg UTI group was smaller than in the 100,000 U/kg UTI and saline control groups (P < 0.05). Treatment with 300,000 U/kg UTI showed a trend to improve neurologic outcome but did not reach statistical significance (P = 0.0693). The significant decrease in neutrophil infiltration was observed in the ischemic hemisphere treated with 300,000 U/kg UTI compared with saline control (P < 0.05). Nitrotyrosine deposition in the ischemic hemisphere was significantly reduced in the 300,000 U/kg UTI group compared with saline control and 100,000 U/kg UTI groups (P < 0.05). CONCLUSIONS: Intravenous pretreatment with 300,000 U/kg UTI reduces focal ischemia-reperfusion injury in the rat brain, potentially opening a novel therapeutic avenue for the treatment of cerebral ischemia. PMID- 12552208 TI - Nociception in persistent pancreatitis in rats: effects of morphine and neuropeptide alterations. AB - BACKGROUND: Most animal models of pancreatitis are short-lived or very invasive. A noninvasive animal model of pancreatitis developed in highly inbred rats by Merkord with symptoms persisting for 3 weeks was adopted in the current study to test its validity as a model of visceral pain in commercially available rats. METHODS: The persistent pancreatitis model was established by tail vein injection of dibutyltin dichloride. Animals were given 10% alcohol in their drinking water to enhance the pancreatitis attack. Blood serum pancreatic enzymes and nociceptive state were monitored for 3 weeks after dibutyltin dichloride or vehicle. Behavioral testing included reflexive withdrawal to mechanical and thermal stimulation of the abdominal area. The effect of morphine on nociceptive behaviors was tested. Histologic analysis of the pancreas and immunohistochemical analysis of substance P and calcitonin gene-related peptide in the spinal cord are included in the study. RESULTS: Compared with naive and vehicle-only injected control groups, rats receiving dibutyltin dichloride demonstrated an increase in withdrawal events after von Frey stimulation and decreased withdrawal latency after thermal stimulation, signaling a sensitized nociceptive state through 7 days. These pain-related measures were abrogated by morphine. Blood serum concentrations of amylase and lipase as well as tissue inflammatory changes and substance P were also significantly elevated during this same time period. CONCLUSIONS: These results indicate that animals with the dibutyltin dichloride induced experimental pancreatitis expressed serum, histologic, and behavioral characteristics similar in duration to those present during acute attacks experienced by patients with chronic pancreatitis. These findings and responsivity to morphine suggest the utility of this model developed in a commercially available strain of rats for study of persistent visceral pain. PMID- 12552210 TI - Antinociceptive effect of low-dose intrathecal neostigmine combined with intrathecal morphine following gynecologic surgery. AB - BACKGROUND: The purpose of this study was to determine whether combination of 1-5 microg intrathecal neostigmine would enhance analgesia from a fixed intrathecal dose of morphine. METHODS: A total of 60 patients undergoing gynecologic surgery were randomized to one of five groups. Patients received 15 mg bupivacaine plus 2 ml of the test drug intrathecally (saline, 100 microg morphine, or 1-5 microg neostigmine). The control group received spinal saline as the test drug. The morphine group received spinal morphine as test drug. The morphine + 1 microg neostigmine group received spinal morphine and 1 microg neostigmine. The morphine + 2.5 microg neostigmine group received spinal morphine and 2.5 microg neostigmine. Finally, the morphine + 5 microg neostigmine group received spinal morphine and 5 microg neostigmine. RESULTS: The groups were demographically similar. The time to first rescue analgesic (minutes) was longer for all patients who received intrathecal morphine combined with 1-5 microg neostigmine (median, 6 h) compared with the control group (median, 3 h) (P < 0.02). The morphine group (P < 0.05) and the groups that received the combination of 100 microg intrathecal morphine combined with neostigmine (P < 0.005) required less rescue analgesics in 24 h compared with the control group. The incidence of perioperative adverse effects was similar among groups (P > 0.05). CONCLUSIONS: The addition of 1-5 microg spinal neostigmine to 100 microg morphine doubled the duration to first rescue analgesic in the population studied and decreased the analgesic consumption in 24 h, without increasing the incidence of adverse effects. The data suggest that low-dose spinal neostigmine may improve morphine analgesia. PMID- 12552209 TI - Prospective study on incidence and functional impact of transient neurologic symptoms associated with 1% versus 5% hyperbaric lidocaine in short urologic procedures. AB - BACKGROUND: The objectives of this study were to compare the incidence, onset, duration and pain scores of transient neurologic symptoms (TNS) with 1% versus 5% hyperbaric lidocaine in spinal anesthesia for short urological procedures in a large prospective study. This study would also evaluate patient satisfaction, and impact of TNS on functional recovery to assess the clinical significance of TNS. METHODS: This was a multicenter, double-blind, randomized controlled trial. Four hundred fifty-three patients undergoing short transurethral procedures were randomized to receive 1% or 5% hyperbaric lidocaine. Eighty milligrams of 1% or 5% hyperbaric lidocaine was administered. During the first 3 days after surgery, the presence of TNS, its intensity and duration, and patient functional level were recorded. An intention-to-treat analysis was used. RESULTS: There was no difference in the incidence of TNS (21% vs. 18%) between 1% versus 5% lidocaine. Patients with TNS had significantly higher pain scores (5.3 +/- 3 vs. 2.3 +/- 3) than patients without TNS during the first 24 h. This difference in pain scores persisted until 72 h postoperatively. There was a significant difference in the daily activities functional scores (2.2 +/- 1 vs. 1.4 +/- 0.8) of TNS non-TNS patients during the first 24 h postoperatively. CONCLUSIONS: There was no difference in the incidence of TNS between the 1% versus 5% spinal lidocaine groups. Pain scores were higher in patients with TNS than those who did not have TNS. During the first 48 h postop, a small proportion of patients who had TNS experienced functional impairment of walking, sitting, and sleeping. PMID- 12552211 TI - High spinal anesthesia for cardiac surgery: effects on beta-adrenergic receptor function, stress response, and hemodynamics. AB - BACKGROUND: This double-blind, randomized, controlled trial examined the effect of high-dose intrathecal bupivacaine in combination with general anesthesia on atrial beta-adrenergic receptor function, the stress response, and hemodynamics during coronary artery bypass graft surgery. METHODS: Thirty-eight patients were randomized to either control (n = 19) or intrathecal bupivacaine (ITB) groups (n = 19). Patients in the ITB group received 37.5 mg intrathecal hyperbaric bupivacaine before induction of general anesthesia. Control patients received an injection of local anesthetic into the skin and subcutaneous tissues (sham spinal). Comparisons were made between groups with respect to atrial receptor desensitization and down-regulation, in addition to circulating catecholamines and hemodynamics. RESULTS: In patients with cardiopulmonary bypass (CPB) times in excess of 1 h, the ITB group had significantly less atrial beta-receptor dysfunction, as measured by maximal isproteronol, 50% maximal isoproterenol, sodium fluoride-stimulated activity, and zinterol stimulation assays of adenylyl cyclase activity (P < or = 0.02) and beta-adrenergic receptor density (P = 0.02). Serum epinephrine, norepinephrine, and cortisol concentrations were significantly lower in the ITB group, independent of CPB times (P < 0.0001, P < 0.001, and P < 0.05, respectively). ITB patients had a higher cardiac index and a lower pulmonary vascular resistance index in the post-CPB time period (P < 0.01 and P < 0.05, respectively). In the pre-CPB period, mean arterial pressure and systemic vascular resistance index were significantly lower in the ITB group. CONCLUSIONS: High-dose intrathecal bupivacaine, when combined with general anesthesia, resulted in less beta-receptor dysfunction and a lower stress response during coronary artery bypass graft surgery. PMID- 12552212 TI - Spinal anesthesia: functional balance is impaired after clinical recovery. AB - BACKGROUND: The ability of patients to walk without assistance after spinal anesthesia is a determining factor in the time to discharge following ambulatory surgery. The authors compared clinical markers of gross motor recovery with objective data of functional balance after spinal anesthesia. METHODS: Twenty-two male patients with American Society of Anesthesiology physical status I or II who were scheduled for perineal surgery were studied during recovery from spinal anesthesia to compare the predictive accuracy of clinical markers of ambulatory readiness (e.g., full knee flexion and extension) with that of an objective method of measurement focused on functional balance. Lumbar puncture was performed at the L2-L3 or L3-L4 interspace using a 25-gauge Whitacre needle, with patients in the sitting position. A 3-ml mixture of 5 mg bupivacaine (heavy) and 10 microg fentanyl was injected. Block regression and restoration of motor function were assessed and recorded. Functional balance was measured using a computerized force platform method. RESULTS: The majority of patients maintained motor function and proprioception sensation at the onset of surgical anesthesia, as indicated by performance on clinical tests of function: 96% were able to perform the straight leg increase; 82, 77, and 91%, respectively, were able to perform full knee flexion and extension, perform heel-to-shin maneuvers, and identify joint position in the supine position. Postoperatively, clinical return of motor function occurred much earlier than recovery of functional balance. At 60 min after onset of spinal anesthesia, 22 patients (100%) had recovered sensory and gross motor function, but only 36% could stand, and 8% could walk without assistance (P < 0.01). At 150-180 min after onset, 96-100% of patients achieved the levels of functional balance that permitted adequate ambulation. CONCLUSIONS: The results suggest that the recovery time to unassisted ambulation is longer than has been assumed, and that the standard clinical markers of gross motor function are poor predictors of functional balance following ambulatory surgery. PMID- 12552213 TI - Indocyanine green: evidence of neurotoxicity in spinal root axons. AB - BACKGROUND: The inadvertent intravascular injection of a local anesthetic during epidural anesthesia is an uncommon but potentially serious complication. Epinephrine, the most commonly used marker, does not provide sufficient sensitivity to exclude intravascular injection in all patient populations. The dye indocyanine green (ICG) has been proposed as an alternative marker. It has been demonstrated that ICG could be used to detect intravascular injections with a simple transcutaneous spectrophotometric technique. Although the safety of intravenous ICG is well documented, its neurotoxic potential requires careful study given the probability of inadvertent intrathecal injection during test injections used to verify epidural catheter placement. METHODS: In this study, the authors investigated the neurophysiologic effects of clinically relevant concentrations of ICG (range, 28.6-286 microm) on single myelinated and unmyelinated dorsal root axons in rats by measuring effects on impulse generation and conduction. RESULTS: In contrast to the apparent absence of toxicity when injected intravenously, ICG applied to intact dorsal roots at concentrations likely to be encountered with an epidural test dose produced long-lasting conduction block (21 of 26 axons) or spontaneous bursting activity (7 of 26 axons) in myelinated and unmyelinated dorsal root axons. CONCLUSION: Given this apparent neurotoxicity, ICG should not be used when intrathecal or nerve root injection is possible. PMID- 12552214 TI - Activation of peripheral excitatory amino acid receptors decreases the duration of local anesthesia. AB - BACKGROUND: Postsurgical wound infiltration with the -methyl-d-aspartate receptor antagonist ketamine and bupivacaine can significantly prolong the duration of local anesthesia. One possible mechanism for this effect is that increased glutamate concentrations, caused by tissue damage, sensitize nociceptive primary afferent fibers through activation of peripheral excitatory amino acid receptors. METHODS: The effect of intramuscular injection of hyper-tonic glutamate (1,000 mm), dextrose (1,400 mm), glutamate (1,000 mm) with the broad spectrum excitatory amino acid receptor antagonist kynurenate (100 mm), or isotonic saline (155 mm) on the duration of masseter muscle afferent fiber blockade after lidocaine (37 mm [1%], 10 microl) infiltration, on muscle edema formation and on muscle blood flow was examined. RESULTS: Injection of either glutamate or dextrose significantly shortened the duration of lidocaine blocks compared with isotonic saline; however, block duration was significantly shorter after glutamate than after dextrose. Injection of glutamate, but not isotonic saline, dextrose, or glutamate with kynurenate, significantly decreased the mechanical threshold of muscle afferent fibers. Injection of glutamate, dextrose, or glutamate with kynurenate produced equivalent large, long-lasting (> 60 min) edemas with high initial peak extracellular water content. Peak extracellular water decreased more rapidly when kynurenate was coinjected with glutamate. Both glutamate and dextrose significantly increased muscle blood flow for 30 min after injection. Glutamate induced increases in blood flow were attenuated by kynurenate. CONCLUSIONS: These results suggest that shortened lidocaine block durations observed after glutamate injection into the masseter muscle result from sensitization of afferent fibers as well as increases of peak extracellular water content and blood flow in masseter muscle. These effects of glutamate are mediated in part through activation of peripheral excitatory amino acid receptors. PMID- 12552215 TI - Postoperative nausea and vomiting in regional anesthesia: a review. PMID- 12552216 TI - Computational aspects of anesthetic action in simple neural models. PMID- 12552217 TI - Perioperative considerations in the patient with a left ventricular assist device. PMID- 12552218 TI - Opioid "holiday" following antagonist supported detoxification during general anesthesia improves opioid agonist response in a cancer patient with opioid addiction. PMID- 12552219 TI - Suxamethonium and donepezil: a cause of prolonged paralysis. PMID- 12552220 TI - Dexmedetomidine to facilitate drug withdrawal. PMID- 12552221 TI - A combustive destruction of expiration valve in an anesthetic circuit. PMID- 12552222 TI - Transient cross-resistance to neuromuscular blocking agents in a patient with tetanus. PMID- 12552223 TI - Extensive retroperitoneal hematoma without neurologic deficit in two patients who underwent lumbar plexus block and were later anticoagulated. PMID- 12552224 TI - Paresthesia but no motor response: what's going on? PMID- 12552225 TI - Motor response following paresthesia during interscalene block: methodological problems may lead to inappropriate conclusions. PMID- 12552226 TI - What happened to the paresthesia? PMID- 12552227 TI - 2-octyl cyanoacrylate glue for the fixation of continuous peripheral nerve catheters. PMID- 12552228 TI - A simple arm positioning aid for fracture table cases. PMID- 12552230 TI - Incubation of porcine iris-ciliary bodies to study the mechanisms by which nitric oxide donors lower intraocular pressure. AB - BACKGROUND: We previously reported that several nitric oxide (NO) donors, guanylate cyclase activators, and cyclic GMP lower intraocular pressure (IOP) in rabbits. MATERIAL/METHODS: This study evaluated a novel method for studying cGMP production in the iris-ciliary body after the administration of different NO donors and guanylate cyclase activators. Tissue samples of porcine iris-ciliary body were incubated for 30 or 60 minutes with the test compounds and with or without the phosphodiesterase inhibitor zaprinast. The concentration of cGMP in the iris-ciliary body as an indicator of soluble guanylate cyclase activation was measured by radioimmunoassay. RESULTS: The tested NO donors - SNOG, NONOate, NOR 3, and SNAP - were shown to release NO in incubation medium, and clearly increase cGMP concentration in the iris-ciliary body. Cyclic GMP production was 2-5 times higher with nitrosocaptopril and about 10 times higher with SNP than in the unstimulated control tissue incubation. Captopril, the reference for nitrosocaptopril, did not induce cGMP production in the porcine iris-ciliary body. ODQ, a guanylate cyclase inhibitor, shut down the production of cGMP after the administration of nitrosocaptopril and SNP. The guanylate cyclase activators YC-1 and atriopeptin III increased cGMP dose-dependently. CONCLUSIONS: In this novel tissue incubation method, several NO donors and guanylate cyclase activators increased cGMP production in the porcine iris-ciliary body. This method can be used to screen new molecules in terms of cGMP production, since the ciliary body is important in lowering intraocular pressure. PMID- 12552231 TI - The influence of acute hypercapnia on the quantity and oxidative metabolism of bronchoalveolar lavage-derived leukocytes in the mechanically ventilated rabbit. AB - BACKGROUND: Permissive hypercapnia exerts a protective influence in tissue injury, possibly by attenuating free radical activity. Increased CO2 concentrations may decrease the production of phagocyte reactive oxygen intermediates (ROI). In this study, we used bronchoalveolar lavage (BAL) to determine whether isooxic permissive hypercapnia due to hypoventilation would influence neutrophil recruitment into alveolar spaces and ROI production of BAL phagocytes, as measured by luminol dependent chemiluminescence. MATERIAL/METHODS: 27 Chinchilla rabbits randomized into three groups: control group (C; n=7), normocapnia (NC; n=10), and hypercapnia (HC; n=10) were anaesthetized with pentobarbitone and intubated. Bronchoalveolar lavage was obtained either after intubation (controls) or after mechanical ventilation lasting 240 minutes under normocapnia or permissive hypercapnia conditions. BAL cells were counted and luminol-dependent chemiluminescence estimations were performed. RESULTS: The BAL neutrophil count was significantly higher in the hypercapnia group than in normocapnia animals. The chemiluminescence of opsonized zymosan-stimulated cells was significantly higher in the normocapnia group than in the controls; the differences between the HC and NC animals, and between the HC and control animals, did not reach statistical significance. CONCLUSIONS: In the mechanically ventilated rabbit under pentobarbitone anaesthesia, acute isooxic hypercapnia due to hypoventilation increases neutrophil BAL count, without direct impact on mixed BAL-cell chemiluminescence; moreover, conventional ventilation increases the chemiluminescence of opsonized zymosan-stimulated BAL cells PMID- 12552232 TI - Age-dependent increase of 8-oxoguanine-, hypoxanthine-, and uracil- DNA glycosylase activities in liver extracts from OXYS rats with inherited overgeneration of free radicals and Wistar rats. AB - BACKGROUND: Oxygen free radicals have been hypothesized to play an important role in the process of aging. MATERIAL/METHODS: To investigate the correlation between oxidative stress and accumulation of DNA damage we determined age-dependent levels of activities eliminating 8-oxoguanine, hypoxanthine and uracil from DNA in liver cells from OXYS rats, which are characterized by inherited overgeneration of free radicals, in comparison with those of control Wistar rats. RESULTS: A pronounced difference in the specificity of mitochondrial and nuclear 8-oxoguanine DNA glycosylase/AP lyase activities were revealed in both cases. Our results suggest the induction of an 8-oxoG-, uracil- and hypoxanthine-specific repair pathway with age in both types of rats. The levels of 8-oxoguanine DNA glycosylase/AP lyase activities in nuclear extracts from both strains of rats are comparable and approximately tenfold higher than in mitochondrial extracts. On the contrary, 8-oxoguanine DNA glycosylase/AP lyase activity in OXYS mitochondrial extracts was remarkably higher than that from old Wistar rats, and a significant increase of this activity occurs earlier in OXYS than in Wistar rats. CONCLUSIONS: Our results are consistent with the shorter life-span of OXYS rats, and with the mitochondrial theory of aging, which postulates that the accumulation of DNA damage in mitochondrial genomes leads to mitochondrial dysfunction and accelerates the process of aging. PMID- 12552233 TI - Analysis of estrogen receptor (ERalpha and ERbeta) and progesterone receptor (PR) polymorphisms in uterine leiomyomas. AB - BACKGROUND: Leiomyomas are the most common tumors in women, found in up to 30% of women in active reproductive life. These tumors are estrogen- and progestin responsive. In fact, they do not occur before menarche, undergoing rapid increase in size during pregnancy with consequent fetal wastage. Conversely, they can regress or even calcify after menopause or castration. Given the central role of sex steroids in uterus trophism, estrogen and progesterone receptors are important mediators of hormonal bioeffects, and the genes encoding these receptors become easy candidates for uterine proliferative disorders. MATERIAL/METHODS: We examined the distribution of common polymorphisms of ERalpha, ERbeta and PR genes in 413 Caucasian females: 225 post-menopausal healthy controls and 188 premenopausal women affected by uterine leiomyomas. The polymorphisms were evaluated both in constitutive and tissue DNA by PCR amplification, specific endonuclease digestion, and then detection with agarose or polyacrylamide gel electrophoresis. RESULTS: The observed allele frequencies did not deviate from the expected Hardy-Weinberg distribution in our population. In case-control analysis, no variant frequency of the studied genes differed significantly between control subjects and patients. CONCLUSIONS: Polymorphisms in the genes encoding for ERalpha, ERbeta and PR did not correlate with the occurrence of uterine leiomyomas in our Caucasian population. PMID- 12552235 TI - Pharmacokinetics studies and toxicity profile of raltitrexed used by intraperitoneal route in normothermia in a pig model. AB - BACKGROUND: To define the pharmacokinetic and toxicity profile of raltitrexed administered by intraperitoneal route in a normothermic pig model. MATERIAL/METHODS: Twenty-one female pigs were divided in three groups. Under general anesthesia, pigs in groups 1 and 2 underwent laparotomy with resection of a small hepatic wedge, bowel anastomosis and portal and systemic vein catheterization. This was followed by an intraperitioneal delivery of raltitrexed: a dose of 1 mg was given to group 1 pigs, whereas group 2 received 2 mg. Serial sampling of portal blood, systemic blood and peritoneal fluid was then undertaken in both groups for pharmacokinetics studies. Pigs were followed daily for a period of 10 days after surgery with recordings of signs of toxicity. On day 10, an exploratory laparotomy was undertaken on each pig in order to assess peritoneal toxicity after which they were euthanized. Pigs in group 3 were used for bioavailability evaluation. RESULTS: Three deaths were recorded within 24 hours of surgery; a technical problem was identified in all cases and no deaths were the results of raltitrexed toxicity. A peritoneo-plasma gradient of 100:1 was obtained. Cmax in plasma were of 28 ng/ml and 54 ng/ml for group 1 and 2, respectively. Tmax were 180 min for both groups. AUCplasmatic was double in the 2 mg group compared to the 1 mg group. CONCLUSIONS: Raltitrexed administered by IP route in pigs is non-toxic. Pharmacologically, there are few interindividual variations and the small first-past effect did not significantly alter the high peritoneo-plasmatic gradient PMID- 12552234 TI - Aminopeptidase activity in renovascular hypertension. AB - BACKGROUND: Renovascular hypertension is accompanied by increased renin angiotensin system activity. Angiotensin II (Ang II) is degraded by aminopeptidases into various metabolites. Increased Ang II production and decreased Ang II degradation may have pathological consequences in maintaining high tissue/plasma Ang II levels. MATERIAL/METHODS: We report the effects of renovascular hypertension on alanyl- (AlaAP), arginyl- (ArgAP), cystinyl- (CysAP), aspartyl- (AspAP), glutamyl- (GluAP) and pyroglutamyl- (pGluAP) aminopeptidases, using arylamides as substrates. The enzymatic activities were analyzed in plasma, right atrium, lung, left ventricle and aortic ring of rats, normotensive (sham-operated) and hypertensive (Goldblatt two-kidney one-clip, G2K1C), treated or not with the AT1 receptor antagonist valsartan. All determinations were performed six weeks after surgery. RESULTS: Whereas the atrium exhibited an increase, the lung, ventricle and aorta showed a decrease of aminopeptidases in G2K1C rats. Except in the aorta of normotensive rats, valsartan did not affect aminopeptidases in the groups studied. CONCLUSIONS: The present study may imply reduced metabolism of angiotensin II in the lung and aorta of G2K1C rats. This down-regulation could prolong the half-life of Ang II and contribute to the maintenance of hypertension. Changes in AP activities did not appear to be part of the action mechanism of AT1 receptor blockade in hypertensive rats PMID- 12552236 TI - Experimental studies of serum cytokine concentration following pancreatic electrolytic ablation. AB - BACKGROUND: Pancreatic cancer has a poor prognosis, with symptomatic palliation being the priority in the majority of cases. Alternative palliative techniques, such as local ablation, are under investigation. Palliative per-ductal electrolytic ablation of the pancreas has been reported, in an experimental series. It is important to establish that this technique is associated with low morbidity and mortality if it is to be acceptable. The 'systemic inflammatory response syndrome' (SIRS) is a recognised and serious complication of both acute pancreatitis, and certain locally ablative techniques. This study aimed to determine whether pancreatic electrolytic ablation is associated with an increase in serum IL-1b and TNF-a concentrations, these cytokines playing integral roles in the inflammatory pathway of SIRS. MATERIAL/METHODS: Eight pigs underwent per ductal electrolytic ablation of the pancreas. Serum samples, taken pre operatively and post-operatively for two weeks, were analysed for IL-1b and TNF-a concentrations. Variations in cytokine levels were statistically analysed. RESULTS: Post-operative serum IL-1b and TNF-a concentrations did not significantly increase on pre-operative figures. There were no other clinical, biochemical or histological indicators of a SIRS-like phenomenon. CONCLUSIONS: The results from this study suggest that electrolytic ablation of normal pancreas is not associated with either an increase in serum concentrations of IL-1b and TNF-a or a SIRS phenomenon. Although this study has only investigated the serum levels of two cytokines, IL-1b and TNF-a have both been shown to have a central role in the inflammatory cascade that leads to SIRS. PMID- 12552237 TI - The relationship between eruption and length of mandibular incisors in young rats. AB - BACKGROUND: Rat incisor teeth exhibit continuous growth, hence they have frequently been used to study the eruption process. The aim of our study was to investigate changes in the eruption rate of impeded and unimpeded rat incisors and to determine if any relationship existed between eruption rate and length of the clinical crown. MATERIAL/METHODS: An image analysis technique was used to measure the length of the mandibular incisors of 23 young female Sprague-Dawley rats, aged 7 to 8 weeks. Over a four-week period, the mandibular right incisors of the 11 rats in the experimental group were shortened every two or three days. Another group of 12 rats served as the control. RESULTS: The mean unimpeded eruption rate was 1.0+/-0.1 mm/day, while the impeded eruption rate was 0.6+/-0.1 mm/day. On day 2, the eruption rate of the unimpeded mandibular incisors accelerated to 163.9% when compared to the controls. The impeded mandibular incisors increased in length by 8.8% and 12.1% for the experimental and control groups respectively. An inverse relationship was identified between the eruption rate and the length of clinical crown of the impeded mandibular incisors, while the unimpeded eruption rate was unrelated to the length difference between the left and right incisors if the length difference exceeded 0.5 mm after accelerated eruption. CONCLUSIONS: It was found that the mandibular incisors of young rats need to be shortened every two days in order to study unimpeded tooth eruption, and the shorter the tooth is cut the higher the rate of eruption. PMID- 12552238 TI - Biophysical characterization of an integrin-targeted non-viral vector. AB - BACKGROUND: The formulation of polycationic complexes containing plasmid DNA for optimal transfection in vitro and in vivo for DNA vaccination, gene therapy and other applications continues to be a major research goal. Here we present new data on the biophysical properties of an integrin-targeted plasmid DNA (LID) formulation. MATERIAL/METHODS: Two plasmids (D), pEGFP (4.7 kb) and pCI-luc (5.7 kb), were mixed with a synthetic a5b1 integrin-targeted peptide (I), [K]16 GACRRETAWACG, in the presence of a cationic liposome (L), Lipofectin, composed of DOTMA and DOPE to form LID complexes. The physical properties of the complexes were measured using a variety of techniques including dynamic light scattering and fluorescence methods. The in vitro gene delivery to neuroblastoma cells with LID complexes was also assessed. RESULTS: We demonstrate the effects of complex size and charge ratio on in vitro transfection of mouse (Neuro-2A) and human (IMR 32) neuroblastoma cells. We report a significant increase in the level of luciferase and green fluorescent protein expression when transfection is performed in buffers of physiological ionic strength and hypothesise that the enhancement in transfection is caused by an increase in the size of the complexes observed during mixing and maturation. CONCLUSIONS: Cell transfection is also shown to be dependent on complex size and charge ratio, with large complexes prepared at charge ratios above 4.0 demonstrating efficient transfection. PMID- 12552239 TI - Hsp70 is present in human saliva. AB - BACKGROUND: There is increasing evidence that chaperones are also present outside the cell, exerting cytokine-like effects and influencing immune recognition. Hsp70 has been found to be present in human blood sera. Chaperonins Cpn10 and Cpn60 are present in pancreatic juice, but Hsp70 is not. These observations raise the possibility that molecular chaperones may be present in other secretory fluids, such as human saliva. MATERIAL/METHODS: Human whole saliva was collected from six participants under resting conditions and secretory stimulation. The samples were precleared by centrifugation and sterile filtered. Salivary volume, protein concentration and amylase activity were determined. For detection of Hsp70 saliva proteins were separated on a 12.5% SDS-PAGE. Semi-dry Western blot analysis was used with a primary antibody against the inducible form of Hsp70. Hsp70 bands were detected with a horseradish peroxidase-linked secondary antibody and ECL-Western blotting analysis. RESULTS: A single band was recognized around 70 kDa in the saliva of all the participants. There was a significant decrease of Hsp70, and a non-significant decrease of total protein concentration during stimulation, whereas the activity of salivary amylase increased significantly. Stimulation significantly increased the Hsp70, total protein and amylase outputs as well as the amylase/protein ratio, and decreased the Hsp70/amylase and Hsp70/protein ratios. CONCLUSIONS: Hsp70 is secreted to saliva, but unlike amylase is not transported by the exocytotic secretory mechanisms of acinar cells. Passive transport mechanisms of Hsp70 from blood serum or from salivary gland cells may be major routes of salivary Hsp70 secretion. PMID- 12552240 TI - The influence of etoposide on the distribution of tubulin in human leukemia cell line HL-60. AB - BACKGROUND: In the last decade there has been increasing interest in cytoskeletal research in relation to important process of cell biology. However, after decades of debate many issues remain unsettled. The aim of our study was to estimate the distribution of tubulin in etoposide-induced HL-60 cells and address the question of whether the reorganization of tubulin may be involved in apoptosis. MATERIAL/METHODS: HL-60 cells were incubated with etoposide (0.02-200 mM/L) for 72 hours. Tubulin was estimated by the indirect immunofluorescence method. TRITC labeled goat anti rabbit IgG was used to label tubulin. RESULTS: Significant changes occurred in cell morphology of and tubulin organization at 200 mM/L etoposide: large cells, brightly labeled for tubulin, in cytoplasm. At 200 uM/L etoposide, we also observed an increase in apoptotic cells with bright staining for tubulin at the site of apoptotic body formations. Cells with features of apoptosis were frequently seen also at 20 mM/L etoposide. The exposure of cells to 0.02, 0.2, or 2 mM/L etoposide did not result in any visible changes in morphology or tubulin organization. Cell size, the number of viable cells, and the percentage of apoptotic cells correlated significantly with the dose of etoposide (p<0.001). CONCLUSIONS: These results indicate that etoposide promotes changes in the distribution of tubulin in HL-60 cells, depending on the dose, and that the reorganization of tubulin seems to be involved in apoptosis PMID- 12552241 TI - Serum neutrophil elastase levels predict initial clinical condition but do not correlate with the progression of interstitial lung disease. AB - BACKGROUND: Recent publications have suggested that neutrophil elastase (NE) may have a role in evaluating the clinical condition of patients with interstitial lung diseases (ILD). This study aims to evaluate the role of serum NE levels in the follow-up of patients with ILD. MATERIAL/METHODS: A group of 100 consecutive patients diagnosed with various ILDs were prospectively studied on two successive visits. On the first visit, the clinical condition of each patient was assessed, and blood count, pulmonary function tests, chest x-ray and serum NE levels (by latex agglutination assay) were performed on all patients. On the second visit, 8 months later, the patients were classified in two groups: those with unfavorable progression and those who were either in the same clinical status or showed good progression. RESULTS: There was a weak correlation between NE and age (r= -0.383; p < 0.0005). Sex, age, NE and the treatment received were found to be independent predictors of the initial clinical condition. Multivariate analysis including these variables demonstrated that higher levels of serum NE predicted the worst clinical presentation (odds ratio: 4.392; 95% CI: 1.665 - 11.586; p = 0.003). However, none of the variables were found to be significantly different when the progression of the disease was assessed. CONCLUSIONS: Although NE seems to be a good marker for the initial clinical condition in this group of diseases, its role as a prognostic factor could not be proven PMID- 12552242 TI - Impaired hepatic mitochondrial oxidation using the 13C-methionine breath test in patients with macrovesicular steatosis and patients with cirrhosis. AB - BACKGROUND: In cirrhosis, hepatic mitochondriae exhibit morphological and functional abnormalities. In human steatosis, ultrastructural changes are reported in the absence of ethanol. Routine evaluation of mitochondrial function is difficult. We used a breath test to explore hepatic mitochondrial oxidation in vivo. MATERIAL/METHODS: The 13C-methionine breath test was performed in healthy subjects (n=15), patients with cirrhosis (n=25), and patients with biopsy-proven severe (> 40% involved hepatocytes), non-alcoholic macrovesicular steatosis (n=18). After oral administration of 13C-methionine (200 mg), hepatic mitochondrial decarboxylation was measured by breath [13C]-C02 enrichment, expressed as dose/h and delta over base (DOB), for 1 hour, by isotope ratio infrared spectroscopy. Results were normalized against BMI. RESULTS: At 60 minutes, patients with steatosis had reduced exhalation of 13C02 as compared to healthy subjects (dose/h: -47%, 18.8+/-12 vs 36+/-6.1; DOB: -52%, 40.4+/-32 vs 85+/-20, p<0.05). Cirrhotics had even lower values as compared to patients with steatosis (dose/h: -60%, 7.5+/-3.4 vs 18.8+/-12, p<0.05). In cirrhosis, dose/h correlated (r=0.68) to aminopyrine breath test values, a microsomal function test, and was inversely correlated (r=-0.48) to the Child-Pugh score. CONCLUSIONS: Hepatic mitochondrial oxidation as reflected by the 13C-methionine breath test is impaired both in patients with pure non alcohol-related severe macrovesicular steatosis and in patients with cirrhosis of mixed etiologies. This non- invasive test can be used to monitor hepatic mitochondrial function in vivo. PMID- 12552243 TI - Helicobacter pylori aeropositivity and the incidence of hyperemesis gravidarum. AB - BACKGROUND: We studied the incidence of Helicobacter pylori infection in patients with hyperemesis gravidarum (HG) in comparison with non-HG pregnant women, and investigated whether there was a correlation between positive serology for H. pylori IgG, the time of onset, and the duration of HG symptoms. MATERIAL/METHODS: 54 pregnant women with HG were enrolled in a matched case - control study. For each case in the group, a pregnant woman without HG matched for age, parity and gestational age was selected as a control. The patients in the experimental group were asked the exact time of symptom onset, and both groups were monitored during pregnancy for symptoms and outcome. Serum H. pylori IgG antibody was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Positive serum H. pylori IgG antibody was detected in 88.9% of the patients in the experimental group vs. 40.7% of the controls (P<0.001). Three of the four patients with abortions in each group were seropositive. Age, parity, level of education, symptom onset, and duration and outcome of pregnancy were comparable in both seropositive and seronegative patients with HG. CONCLUSIONS: Although more patients with HG were seropositive for H. pylori infection than controls, we were not able to demonstrate correlation between seropositivity for H. pylori and the time of onset or duration of HG symptoms. Although H. pylori infection may be an important factor in exacerbating HG, it may not represent the sole cause of the disease. PMID- 12552244 TI - Could incarceration of inguinal hernia in children be prevented? AB - BACKGROUND: The aim of this study was to determine the incidence of incarceration in children with congenital inguinal hernias and to ascertain if incarceration could have been prevented. MATERIAL/METHODS: Records of 1582 children who underwent inguinal herniorrhaphy between 1993 and 2000 were analysed regarding the time and circumstances of hernia incarceration. RESULTS: Incarcerated hernia developed in 153 cases (9.7%) developed, of whom 81 (52.9%) were known to have had prior hernia incarceration and 81 (52.9%) were under one year of age. Manual reduction of hernia was successful in 130 cases (85%), and 99 of them (76.2%) underwent hernia repair during the same stay. Emergency surgery was necessary in a total of 36 children (23.5%), 23 with the first and 13 with the second incarceration episode. Complications occurred in 16 children with incarcerated hernia (10.4%), including intestinal necrosis, omentum and appendix infarction, infarction of the testis, torsion of the ovary, recurrent hernia and wound infection. CONCLUSIONS: The first incarceration of known inguinal hernia could have been avoided in more than half of our patients and the second in one-fourth after successful manual reduction. PMID- 12552245 TI - Procollagen I carboxyterminal propeptide (PICP) as a bone formation marker and carboxyterminal telopeptide of type I collagen (ICTP) as a bone degradation marker in children with chronic renal failure under conservative therapy. AB - BACKGROUND: Biochemical and hormonal disturbances in calcium-phosphate storage accompanying chronic renal failure (CRF) lead to the loss of bone mass and the destruction of bone microarchitecture. The aim of this study was to evaluate the serum levels of PICP (procollagen I carboxyterminal propeptide) and ICTP (carboxyterminal telopeptide of type I collagen) as markers of bone growth in CRF children treated conservatively. MATERIAL/METHODS: 34 children (16 female and 18 males) with predialytic CRF, aged 6 to 18 years (average age 11.3+/-3.8 years) were included in the experimental group. The controls were 20 healthy age-matched children. The experimental subjects were divided into two subgroups, based on the blood level of intact PTH (iPTH). Subgroup Ia (n=7) consisted of children with normal serum concentrations of iPTH. Subgroup Ib (n=27) consisted of children with elevated serum concentrations of iPTH. RESULTS: In group Ia characteristic correlations were found between OST (osteocalcin) and ICTP, and between creatinine and ICTP. In group Ib, positive correlations were found between iPTH and PICP, iPTH and ICTP, OST and ICTP, and ICTP and PICP. In this group no correlation was found between marker turnover and creatinine. CONCLUSIONS: The results of our study of PICP and ICTP as markers of bone metabolism in children with CRF in the predialysis period indicate that their levels should be routinely monitored as specific biochemical parameters of bone structure. PMID- 12552246 TI - Mucosal adenosine deaminase activity and gastritis histology: a comparative study of partially resected and intact stomachs. AB - BACKGROUND: Adenosine deaminase is an enzyme which is postulated to have a role in the generation of gastric mucosal inflammation. The aim of our study was to determine and compare adenosine deaminase activity in the gastric mucosa of patients with chronic gastritis developed in partially resected and intact stomachs. MATERIAL/METHODS: 182 patients were studied, 102 non-operated and 80 after distal gastric resection. Biopsy specimens were taken endoscopically from the gastric mucosa 2 cm proximal to the stoma or corresponding upper third of the intact stomach. Gastritis was classified according to the Sydney system. The activity of adenosine deaminase in the mucosal homogenates was measured by determination of ammonia liberated from the substrate and expressed in nmol NH3/mg protein/min. RESULTS: Adenosine deaminase activity was lower in partially resected than in intact stomachs, regardless of Helicobacter pylori infection. While no difference was found in adenosine deaminase activity between Billroth I and Billroth II procedures in subjects without H. pylori infection, the activity was lower in those with Billroth II procedure in the presence of H. pylori infection. As the severity of gastritis increased, enzyme activity decreased in the mucosa of the intact stomach, but was not significantly altered in the mucosa of the gastric remnant. CONCLUSIONS: Adenosine deaminase activity differs in intact and partially resected stomachs, but it does not appear to be a factor promoting chronic gastritis. PMID- 12552247 TI - Detection of TT virus (TTV) in children and youth with chronic viral hepatitis B and C. AB - BACKGROUND: TTV is a newly discovered virus and little is known about the frequency of TTV infection in children in Poland. The aim of our study was to investigate the frequency of TTV infection in children with chronic viral hepatitis B and C. MATERIAL/METHODS: Two patient groups were tested: 74 patients with chronic hepatitis B aged 4 to 20 years, and 13 patients with chronic hepatitis C aged 10 to 18 years. Nucleic acids were extracted from serum using the spin column technique (QIAGEN(r), Hilden, Germany), and TTV DNA sequences were amplified by nested PCR. RESULTS: TTV DNA was found in 47.3% of patients with chronic hepatitis B and in 53.8% of patients with hepatitis C. Among those patients with chronic B hepatitis there was no statistical difference between the frequency of coinfection with TTV and clinical-histopathological diagnosis, activity of aminotransaminases, frequency of seroconversion of HBeAg to antiHBe, or interferon alpha therapy. CONCLUSIONS: In Poland, TTV viremia is frequent in patients with chronic viral hepatitis B and C. TTV coinfection did not modify the course and activity of chronic hepatitis B or influence the outcome of interferon therapy. PMID- 12552248 TI - Supine body position is an important factor influencing postprandial ambulatory blood pressure. AB - BACKGROUND: Although body position is known to be an important factor influencing the results of ambulatory blood pressure monitoring (ABPM), even very recent studies assessing postprandial blood pressure (BP) have not taken into account the possibility that the examined subjects were lying down after a meal. We addressed this issue by analysing diurnal BP profiles together with subject's reports on supine rest periods and meal consumption during ABPM. MATERIAL/METHODS: The ABPM results of 109 non-medicated subjects were analysed (age 40+/-12 years, daytime BP 132/84+/-15/11 mmHg). BP and heart rate (HR) changes were assessed on the basis of 2-hour means. RESULTS: Subjects who lay down within 2 hours after a meal showed significantly greater decreases in systolic and diastolic BP and HR than individuals who did not: -10+/-10 vs. 1+/ 10 mmHg, -11+/-10 vs. -3+/-7 mmHg, and -7+/-9 vs. 0+/-11 beats/min, respectively. The BP decrease was greater when the duration of supine rest was longer. There were no significant differences between BP/HR changes during those periods when the subjects lay down within 2 hours after the meal and BP/HR changes during those periods when supine rest was not preceded by a meal. The mean changes for the latter group were -9+/-10 mmHg, -11+/-8 mmHg, and -9+/-9 beats/min. for SBP, DBP, and HR, respectively. CONCLUSIONS: Our results indicate that supine body position is an important factor, which may influence postprandial BP investigated by ABPM. Body position should always be taken into consideration when ABPM results are analysed. PMID- 12552249 TI - Cryotherapy in the treatment of angiolymphoid hyperplasia with eosinophilia. AB - BACKGROUND: Angiolymphoid hyperplasia with eosinophilia (ALHE) is a benign and very rare skin disorder of uncertain etiology. It occurs in both dermal and subcutaneous forms, which are regarded as variants of the same condition. Clinically, it is characterised by solitary or multiple, red to brown cutaneous papules or nodules, usually located on the head and neck. ALHE occurs mainly in young to middle-aged adults, and more frequently affects women. CASE REPORT: A 36 year-old Caucasian woman with a one-year history of ALHE is presented. The diagnosis of this rare entity was based on the clinical and histological examination. CONCLUSIONS: Although there are many methods of treatment, the final result is not always satisfactory for patient. We present our case as a reminder that cryotherapy can be regarded as an easy, safe and successful method of treatment in some patients. PMID- 12552250 TI - The in vivo binding site for oncoprotein c-Myc in the promoter for Epstein-Barr virus (EBV) encoding RNA (EBER) 1 suggests a specific role for EBV in lymphomagenesis. AB - BACKGROUND: Epstein-Barr virus (EBV) was isolated in the 1960s from the African childhood tumor, Burkitt's Lymphoma (BL), characterized by the translocation of the c-myc gene into one of the immunoglobulin loci. Due to the extreme discrepancy between the widespread dissemination of EBV infection and the overall rarity of EBV-related tumors, it remains an open question whether EBV is really a human tumor virus, and if so, what specific contribution EBV may have to tumorigenesis. MATERIAL/METHODS: Protein binding at the EBER locus of EBV was analyzed by genomic footprinting electrophoretic mobility shift, reporter gene assay, and chromatin immunoprecipitation in a panel of six B-cell lines. RESULTS: Several novel in vivo protein binding sites were found in the EBER locus. Among those, a prominent binding site, 130 base pairs upstream of the EBER1 gene, contains two E-boxes providing a consensus sequence for binding of the transcription factor and oncoprotein c-Myc to the EBV genome. CONCLUSIONS: Based on the discovery of a binding site for c-Myc in the EBV genome, a new molecular model for the specific role of EBV as a causal factor in the origin of endemic Burkitt's Lymphoma is proposed. Translocated and deregulated c-myc directly activates and maintains the antiapoptotic functions of the EBER locus in a single EBV-infected B cell undergoing the germinal center (GC) reaction. With the balance shifted towards cell survival, the oncogenic potential of the pro apoptotic c-Myc protein is unmasked in the translocated GC cell. This single translocated and surviving cell is the founder cell of an endemic BL. The new model reinstitutes EBV as a real human tumor virus. PMID- 12552251 TI - The evaluation of tonometry and self-tonometry with Ocuton tonometers. AB - BACKGROUND: The purpose of the present study was to evaluate the clinical applicability of the Ocuton-A and Ocuton-S applanation tonometers, and to compare their use and accuracy parameters to those of Goldmann applanation tonometry (GAT). MATERIAL/METHODS: In the first study, intraocular pressure (IOP) was measured with an Ocuton-A tonometer, followed by another measurement using GAT in 15 subjects (30 eyes). Ocuton-tonometry was performed on three occasions separated by three-minute intervals. In Study 2 the impact of increased patient familiarity on measurement accuracy using the self-tonometer was investigated in 5 subjects (10 eyes) by comparing two consecutive series of five Ocuton-S measurements. In Study 3 nine trained volunteers measured their own IOP with an Ocuton-S self-tonometer after GAT and Ocuton-A measurements by an experienced investigator. RESULTS: Study 1 showed that the measured IOP value was significantly higher using the Ocuton-A tonometer, compared to values obtained using GAT. In Study 2 we found that increased practice in self-tonometry did not alter measurement accuracy. In Study 3 the difference between IOP readings obtained with the Ocuton-S and Ocuton-A instruments was not significant. CONCLUSIONS: The portable Ocuton-A tonometer may become a useful instrument for IOP measurement by professional workers outside the clinic. Qualified patients are able to use the Ocuton-S self-tonometer reliably even after limited training, and its measurement accuracy is acceptable for IOP monitoring. Both the Ocuton A and S devices consistently overestimate the IOP by approximately 2 mm Hg compared to the corresponding Goldmann readings. PMID- 12552252 TI - Biological rhythms, endothelial health and cardiovascular disease. AB - The activity of several components of the vascular system appears to be diurnally regulated. Endothelial cell activation, leukocyte and platelet interactions and lipoprotein metabolism have all been shown to vary with time of day, but whether these variations are due to the endogenous circadian clock, exogenous factors, such as the light-dark cycle, or an interaction between the two remains to be determined. Endothelium-dependent vasodilation also varies diurnally. This rhythmicity is lost in individuals with established coronary disease has been shown to occur in the early stages of atherosclerosis. The incidence of coronary events appears to be higher in the early hours of the morning, this may be related to heightened activity of the autonomic nervous system at this time. Higher circulating levels of catecholamines in the morning are associated with increased vascular tone, affecting circulating blood volume and blood pressure. Time dependent variations may be of particular significance for individuals with disrupted circadian rhythms, including rotating shift workers, transmeridian travellers and blind individuals with no light perception. Variations in endothelial function are observed during the menstrual cycle, varying with circulating oestrogen levels. Oestrogen deficiency in postmenopausal women may contribute to endothelial dysfunction, together with other modifiable risk factors. The absolute risk of coronary disease is greater for men than for pre menopausal women. Following the menopause gender differences in coronary risk are thought to diminish, although this remains controversial. This review focuses on the influence of both endogenous biological rhythms and environmental factors on the function and health of the human vascular system. PMID- 12552254 TI - Pathogenetic aspects of immune deficiency associated with beta-thalassemia. AB - Infectious complications constitute the second most common cause of mortality and a main cause of morbidity in beta-thalassemia. Besides the high risk of blood borne infections associated with multiple transfusions, the increased susceptibility of these patients to infectious diseases has been attributed to a coexistent immune deficiency. Immune abnormalities have also been held responsible for the frequent occurrence of malignancies in beta-thalassemia, especially leukemia and lymphomas. Recent studies on immune competence in beta thalassemia have revealed numerous quantitative and functional defects, involving T and B lymphocytes, immunoglobulin production, neutrophils and macrophages, chemotaxis, and phagocytosis, as well as the complement system. Regarding pathogenesis, iron overload, a primary complication of both thalassemia itself and transfusion therapy, is thought to be the main precipitating mechanism, due to the important immunoregulatory properties of iron and its binding proteins; iron excess may derange the immune balance in favor of the growth of infectious organisms. Other factors include multiple transfusions, associated with constant allo-antigenic stimulation, as well as with transmission of immunosuppressive viruses; splenectomy, resulting in increased susceptibility to infections by encapsulated bacteria and to immune system modifications; low levels of zinc, another immune regulator; iron chelation therapy, which predisposes to serious infections by yersinia species; and the circulation of abnormal native thalassemic erythrocytes, forming another permanent immune stimulus. Thus surveillance for infections in patients with beta-thalassemia is crucial, while further studies are warranted on immune function abnormalities and the implicated mechanisms. PMID- 12552253 TI - The controversy surrounding selenium and cardiovascular disease: a review of the evidence. AB - Selenium is an essential trace element that is an integral part of many proteins, with catalytic and structural functions. The antioxidant properties of some selenoproteins, such as glutathione peroxidase, may be particularly important in carcinogenesis and heart disease. The content of selenium in food depends on the selenium content of the soil where the plants are grown or the animals are raised. Moreover, the metabolism of selenium is determined by its dietary form: some forms are better utilized than others. Therefore, wide variations have been found in selenium status in different parts of the world. In animal studies, selenium deficiency is associated with cardiomyopathy and sudden death, as well as reduced T-cell counts and impaired lymphocyte proliferation and responsiveness. Abnormalities in liver function, brain, heart, striated muscle, pancreas and genital tract have also been reported. In humans, selenium deficiency has been implicated in the etiology of cardiovascular disease and other conditions in which oxidative stress and inflammation are prominent features, but there is still only limited evidence from epidemiological and ecological studies for this, and the therapeutic benefit of selenium administration in the prevention and treatment of cardiovascular diseases remains insufficiently documented. Interventions studies are currently in progress to assess the benefits of selenium supplements in primary and secondary prevention of atherosclerosis. The results to date are inconclusive and further controlled trials are needed. PMID- 12552255 TI - Retarded outer segment development in TrkB knockout mouse retina organ culture. AB - PURPOSE: To determine the effects of trkB deficiency in the mouse retina on photoreceptor development and retinal organization, in the absence of confounding systemic effects. METHODS: Newborn mice that carried two null trkB alleles (trkB /-) and their wild type (WT) littermates were used for retinal organ cultures. On Day 21, rod development was assessed histologically in plastic sections (outer segment length) and retinal organization was analyzed using retinal cell-type specific antibodies. Anatomical data obtained from the organ cultures were compared to previously published histological results from in vivo data. RESULTS: (1) Rod outer segment length was significantly shorter in retinas from trkB-/- mice in the presence of normal numbers of rods. (2) No dopaminergic amacrine cells were observed in the knockout retina. (3) Unlike in the in vivo condition, recoverin-positive OFF-cone bipolar cells were present in trkB-/- retinas grown in culture. CONCLUSIONS: (1) These results demonstrate that rod outer segment development is compromised in the absence of trkB in the retina. (2) This study further supports our previous conclusion that the elimination of trkB expression alters rod development, because the presence of trkB receptors within the retina is essential for normal rod maturation and not because of confounding systemic effects. (3) More generally, this study stresses the importance of investigating complex phenotypes in gene knockout mice under conditions that isolate the organ under investigation from unrelated systemic variations. PMID- 12552256 TI - Mutation screening of patients with Leber Congenital Amaurosis or the enhanced S Cone Syndrome reveals a lack of sequence variations in the NRL gene. AB - PURPOSE: To determine if mutations in the retinal transcription factor gene NRL are associated with retinopathies other than autosomal dominant retinitis pigmentosa (adRP). METHODS: Genomic DNA was isolated from blood samples obtained from 50 patients with Leber Congenital Amaurosis (LCA), 17 patients with the Enhanced S-Cone Syndrome (ESCS), and a patient with an atypical retinal degeneration that causes photoreceptor rosettes with blue cone opsin. The 5' upstream region (putative promoter), untranslated exon 1, coding exons 2 and 3, and exon-intron boundaries of the NRL gene were analyzed by direct sequencing of the PCR-amplified products. RESULTS: Complete sequencing of the NRL gene in DNA samples from this cohort of patients revealed only one nucleotide change. The C >G transversion at nucleotide 711 of NRL exon 3 was detected in one LCA patient; however, this change did not alter the amino acid (L237L). CONCLUSIONS: No potential disease causing mutation was identified in the NRL gene in patients with LCA, ESCS, or the atypical retinal degeneration. Together with previous studies, our results demonstrate that mutations in the NRL gene are not a major cause of retinopathy. To date, only missense changes have been reported in adRP patients, and sequence variations are rare. It is possible that the loss of NRL function in humans is associated with a more complex clinical phenotype due to its expression in pineal gland in addition to rod photoreceptors. PMID- 12552257 TI - How safe are our hospitals? PMID- 12552258 TI - Clinical trials in NZ: does anybody know what's going on? PMID- 12552259 TI - Research dishonesty and conflict of interest. PMID- 12552260 TI - Adverse events in New Zealand public hospitals I: occurrence and impact. AB - AIM: To assess the occurrence and impact of adverse events in New Zealand public hospitals. METHODS: Two-stage retrospective review of 6579 medical records, selected by systematic list sample from admissions for 1998 in 13 generalist hospitals providing acute care. After initial screening, medical records were reviewed by trained medical practitioners using a standardised protocol. RESULTS: Except for hospital stay, the sample appeared to be closely representative of New Zealand public hospital admissions for 1998 on key demographic and clinical criteria. The proportion of hospital admissions associated with an adverse event was 12.9% (incidence rate, 11.2%), of which nearly one fifth had occurred outside a public hospital (mainly doctor s rooms, patient s home, rest home, or private hospital). Most adverse events had minor patient impact, with less than 15% associated with permanent disability or death. Hospital workload was strongly affected, however, with adverse events adding an average of over nine days (median 4 days) to the expected hospital stay. There was limited evidence of patterning by diagnostic category. The elderly were disproportionately affected. CONCLUSIONS: The study provides representative base parameters that can contribute to the wider understanding, and potential improvement, of patient safety and the quality of care in New Zealand public hospitals. PMID- 12552261 TI - Tauiwi general practitioners talk about Maori health: interpretative repertoires. AB - AIM: This paper reports findings from a qualitative research project investigating how Tauiwi general practitioners talk about Maori health. Tauiwi is a Maori term for non-Maori New Zealanders. METHODS: The transcripts of interviews with 25 general practitioners from urban Auckland on the topic of Maori health, were subjected to detailed discourse analysis. Through these readings, interpretative repertoires (patterns of language use on particular topics) that participants drew upon in their interviews, were described. RESULTS: We outline the main interpretative repertoires utilised by participants in their talk about Maori health. These include key explanatory forms relating to prevalence and causality of Maori health problems, and rationales for specialised practices when working with Maori. CONCLUSIONS: The repertoires, which are considered to be generalised discursive resources for the construction and elaboration of specific arguments, are oriented to constructions of Maori health that either blame Maori for their plight or justify existing service provision. As such, they are antithetical to arguments for changes to policy and practice that might bring about population-level health gains for Maori. PMID- 12552262 TI - Major inequities between district health boards in referred services expenditure: a critical challenge facing the primary health care strategy. AB - AIMS: This study examines variation between district health boards (DHBs) in expenditure on referred services (ie, pharmaceutical and laboratory services), and compares the gap between budgets as determined by the primary care funding formula and actual expenditure on these services. It also analyses the relationship between population need factors and variation from equity. METHODS: Actual DHB referred services expenditure related to GPs for the period July to November 2001 was obtained from the Ministry of Health and compared with expected expenditure calculated from the funding formula. Percentage difference between actual and expected expenditure was calculated for each DHB. Data were also obtained relating to DHB populations (numbers with community services cards (CSCs), ethnicity, and scores by NZDep96 quintiles) and number of GPs. The data were used to calculate rates for these variables, which were then correlated with percentage variation from equity in DHB referred services expenditure. RESULTS: The analysis showed wide percentage variation from equity between DHBs, with Capital and Coast being 17.5% above and Tairawhiti 23.9% below equity. The analysis also showed a high and significant correlation between this inequity and four measures of disadvantage--population per GP, percentage with CSC, percentage of Maori, and NZDep96 scores--which, together, explained more than 50% of the total variation between DHBs. Population per GP was found to be the most significant predictor of variation. CONCLUSIONS: The inverse care law, ie, that those populations in greatest need are those least likely to receive the services they need, remains a dominant feature of New Zealand s primary care system. This is linked to the gross historical underfunding of access to primary care services. A major redistribution of primary care resources, including GPs, supported by much greater investment in better information and research and development, will be critical to the implementation of the government s Primary Health Care Strategy. PMID- 12552263 TI - Pretibial injury in patients aged 50 years and over. AB - AIMS: To establish the rate of, and identify circumstances surrounding, pretibial injury in patients aged 50 years and over. METHODS: The rate of pretibial injury was estimated from national hospital discharge data from the New Zealand Health Information Service (NZHIS) National Minimum Dataset (1986-1999), and estimates of New Zealand s resident population. Cases accepted by the Accident Rehabilitation and Compensation Insurance Corporation (ACC) (1999) were used to estimate a difference in the number of accepted cases (ie, those that were and were not treated in hospital), and the number of cases resulting in hospital admission. Events surrounding pretibial injuries were identified by surveying 75 patients treated at the Wellington Regional Plastic, Maxillofacial and Burns Unit at Hutt Hospital (January 1999 - November 2000). RESULTS: Estimates of pretibial injury in New Zealand were: annual incidence between 420 and 30 500; standardised rate 0.4-0.7 per 1000 population per year (females 0.6-1.0, males 0.07-0.3); crude rate 33 per 1000 population. About 60% of patients remained hospitalised for 2-14 days. Most of the injured were females (85-90% of NZHIS, 74% of ACC), were aged 70-89 years (68% females/males of NZHIS), and of New Zealand European/Pakeha ethnic origin (90%). Over half of the injury events occurred in the home (eg, caused by household items and furniture). CONCLUSIONS: Pretibial injury is a potential problem for older women in New Zealand. As the result of an ageing population, the annual number of cases is expected to increase. PMID- 12552264 TI - Bilateral pneumothorax: an unusual complication of normal delivery. PMID- 12552265 TI - Continuity through change, a threat or a promise? PMID- 12552266 TI - Electronic publications a myth? PMID- 12552267 TI - Asthma, deprivation and smoking: a small area study in Te Tairawhiti. PMID- 12552269 TI - Adams in wonderland. PMID- 12552270 TI - Ashburton: stay a while, stay forever! PMID- 12552268 TI - Type 1 diabetes: glycaemic control during adolescence. PMID- 12552271 TI - Licking prostate and other cancers and coronary heart disease with an Asian style diet. PMID- 12552272 TI - The emergence of amphetamine use in New Zealand: findings from the 1998 and 2001 National Drug Surveys. AB - AIMS: To examine changes in the use of amphetamines, measure current conditions of supply, and investigate harms related to amphetamine use, in New Zealand. METHODS: National Drug Surveys were conducted in 1998 and 2001. In each survey, a sample of approximately 5500 people aged 15-45 years were asked about their alcohol, tobacco, marijuana, and other drug use, using a Computer Assisted Telephone Interview (CATI) system. In 2001, those who had used stimulants (uppers, speed, amphetamine, methamphetamine) within the previous year were asked a number of new questions about harms experienced from use and present conditions of supply. RESULTS: Last-year use of stimulants increased from 2.9% in 1998, to 5.0% in 2001; use of ice (crystal methamphetamine) increased from 0.1% to 0.9%, while use of marijuana (19.9% and 20.3%) and LSD (3.8% and 3.2%) remained static. Forty one per cent of users thought stimulants were easier to obtain in 2001 compared to a year earlier, and 20% thought the price was lower. One fifth of stimulant users reported harmful effects on 'energy and vitality', while one in ten reported harms to 'financial position', 'health', and 'work opportunities'. CONCLUSIONS: Stimulants moved from being the third most popular illicit drug type in 1998, to the second most popular by 2001. Additional research is required to adequately assess the extent of harms experienced by amphetamine users in New Zealand. PMID- 12552273 TI - The Health Practitioners Competence Assurance Bill. PMID- 12552274 TI - Endoscopic injection therapy for non-variceal upper gastrointestinal bleeding at Auckland Hospital. AB - AIM: The aim of this study was to audit the efficacy of endoscopic injection therapy for non-variceal upper gastrointestinal (GI) bleeding at Auckland Hospital. METHODS: The medical records of 183 patients who had undergone endoscopic injection of adrenaline for non-variceal upper GI bleeding at Auckland Hospital between November 1996 and May 1999 were reviewed. RESULTS: Primary haemostasis was achieved in 177 (96.7%) patients and six (3.3%) patients had early surgery due to failure of injection therapy. The overall rate of rebleeding was 18.6%. The rate of rebleeding according to the stigmata seen at the time of endoscopy was 36.8% for patients showing spurting vessel; 24% for active ooze; 18.2% for visible vessel; and 3.2% for adherent clot. Active spurting vessels seen on endoscopy were associated with higher risk of repeat injection (p <0.01) and death (p <0.001). Sixteen (9%) patients had repeat injection. Haemostasis was achieved in all, and no deaths or surgical interventions were observed in this subgroup. CONCLUSIONS: Endoscopic injection therapy is an effective, simple and cheap first-line therapy for non-variceal upper GI bleeding. The repeat injection also appears to be an effective treatment for patients rebleeding after the initial injection therapy. PMID- 12552275 TI - Delay in seeking medical care for self-detected breast symptoms in New Zealand women. AB - AIMS: To investigate the extent of patient delay in new patients with self detected breast symptoms referred to a specialist breast clinic. To examine the association between delay and sociodemographic factors, the practice of breast self-examination (BSE), how the breast symptom was discovered, and personal experience of breast cancer in family and friends. METHODS: Eighty five women referred to the South Auckland Health Breast Clinic completed a questionnaire and interview after arrival at the clinic and prior to their consultation with the medical specialist. Delay was measured as the time interval between initial self detection of a breast symptom and first contact with a general practitioner (GP) for evaluation of the symptom. Analysis of patient characteristics and delay time was conducted using Pearson product moment correlations, t-test for independent samples and one way analysis of variance (ANOVA) where appropriate. RESULTS: The median delay time between initial breast symptom detection and seeing a GP was 14 days. Of the total sample of 85 women, 40% had seen their doctor within 7 days, 52% within 14 days, 69% within 30 days, and 14% had waited over 90 days. Delay time was not significantly associated with any sociodemographic factors. No difference in delay time was found between the ethnic groups of European, Maori or Pacific women. No difference in delay was shown between those women who performed regular BSE and those who did not. However, women who discovered their breast symptom by chance or through BSE had a shorter delay time than women experiencing breast pain. Although experience of a friend or family member with breast cancer was not significantly associated with delay, there was a trend for women who had a family member with breast cancer to have a longer delay time before seeing their GP. CONCLUSIONS: This study is the first in New Zealand to investigate the extent of patient delay for women with self-detected breast symptoms and the factors influencing this delay. The majority of women saw their doctor within one month of self-detection of a breast symptom, however 14% delayed over three months before seeing their GP. Women who had a family member with breast cancer tended to respond to their breast symptom by delaying seeking medical attention, suggesting that risk perceptions of developing breast cancer may influence delay times in seeking medical help. Results from this study are taken from women referred by their GP to a specialist breast clinic. Certainly, further clarification of the factors that influence delay involving broader patient groups is vital to the development of public education initiatives aiming to encourage women to seek prompt medical evaluation of breast symptoms. PMID- 12552277 TI - Folic acid fortification of foodstuffs. PMID- 12552276 TI - The management and outcome of occluded haemodialysis access: a retrospective audit. AB - AIM: To identify best clinical practice for the management of occluded haemodialysis access. Surgery or percutaneous thrombolysis with or without angioplasty, has been used for the management of clotted haemodialysis access, with variable reported success rates. Concerns over high morbidity rates and delays in achieving satisfactory patent arterio-venous (AV) access, led to a retrospective audit of all patients with occluded haemodialysis vascular access between 1 June 1995 and 30 June 2001. METHODS: Data recorded included type of access, procedure used, outcome, complications and hospital stay. RESULTS: There were 45 episodes occurring in 17 patients. 33 of the 45 episodes occurred in synthetic grafts. Eleven of the 17 patients had multiple episodes (range 2 to 11), nine of whom had synthetic grafts. Forty three of the 45 episodes initially underwent DSA on presentation. There was a low success rate with thrombolysis, with only 20 cases effective in re-establishing dialysis. Surgery revision was required to re-establish effective dialysis in 25 of the 45 episodes. Six of 43 thrombolysis procedures experienced a major complication related to excessive bleeding. Primary patency was slightly better for surgery compared with thrombolysis (4.9 months versus 3.8 months). Temporary catheters were inserted for dialysis in 19 of 45 episodes and remained for a mean of 5.8 days. Four patients had a major episode of catheter-related sepsis. Two patients required admission to the Intensive Care Unit (ICU) for management of their sepsis. Patients who failed thrombolysis and required surgery had a prolonged stay, averaging 8.2 days. This was associated with a marked increase in hospital costs. The average cost for successful thrombolysis was $1976, compared with $5348 where surgery was subsequently required. Costing of surgical intervention alone was similar to that of thrombolysis. CONCLUSION: Surgery with dedicated vascular surgeons remains the safest, most rapid and most effective approach to treating occluded dialysis AV fistulae and grafts. PMID- 12552278 TI - The Otago Medical School pre-clinical curriculum. PMID- 12552280 TI - Selecting New Zealand medical students for the New Zealand medical workforce. PMID- 12552281 TI - Selection of medical students at Otago--time for a change? PMID- 12552282 TI - Picking the winners. PMID- 12552283 TI - Why all workplaces should be smoke free. PMID- 12552284 TI - Parental backgrounds of Otago medical students. AB - AIMS: To investigate the background of medical students admitted to the Otago Medical School (OMS) in terms of their parents' occupations, socioeconomic status and education, and to assess how this has been affected by the introduction of substantial tuition fees. METHODS: A questionnaire was used to collect data on the occupations and education of the parents of all successful New Zealand applicants to the OMS between 1987 and 2000. Trends in the data across the years were examined, and the data compared to the wider New Zealand population. Occupational data were also coded against the 1997 New Zealand Socioeconomic Index of Occupational Status. RESULTS: The distribution of data changed very little from 1987 to 2000. Of 2391 students surveyed, 2353 (98.4%) responded to the questionnaire. Of the respondents, 55.2% had at least one parent with a professional occupation at the time they entered the Medical School, 63.2% had at least one university-educated parent, and 13.1% of students had at least one parent who was a doctor. CONCLUSIONS: In comparison to the wider population, the students surveyed were much more likely to have highly educated, professional parents from higher socioeconomic groups. Although only about one in eight medical students came from medical families, this proportion was much higher than in the wider population. These characteristics changed very little across the 14 years of the study, despite substantial increases in university tuition fees. PMID- 12552285 TI - Audit of a collaborative care model suggests patients with acute myocardial infarction are not disadvantaged by treatment in a rural hospital. AB - AIM: Recent data suggest that improvements in survival post myocardial infarction in urban hospitals have not been matched by rural hospitals. We performed an audit of a collaborative model of care between a rural hospital and its base hospital to see if this was also the case in a rural area of New Zealand, and attempted to identify reasons underlying any discrepancy, should it exist. METHODS: The medical records of all patients with a discharge diagnosis of acute myocardial infarction (AMI) over a five-year period were reviewed and data on management, medication and outcomes collected. RESULTS: 196 patients with confirmed AMI were treated from 1995-1999. There was a documented consultation with a cardiologist in 54% of cases. Seventy one per cent of patients were managed and discharged from the rural hospital, while 23% were subsequently transferred to base hospital. The in-hospital, 30-day and one-year cardiovascular mortality rates were 5.6%, 7.7% and 13.5% respectively. CONCLUSIONS: These figures compare favourably with those previously published in NZ and internationally, suggesting that rural hospitals can achieve similar outcomes to larger centres when working in close collaboration with base hospital specialists. PMID- 12552286 TI - Still dying from second-hand smoke at work: a brief review of the evidence for smoke-free workplaces in New Zealand. AB - AIM: To briefly review the evidence on the hazard from, and control of, second hand smoke (SHS) in New Zealand workplace settings. METHODS: Medline searches and searches of New Zealand health databases for unpublished reports. RESULTS: The New Zealand evidence suggests that over 30% of workers continue to be exposed to SHS in workplace settings. The best available estimate is that SHS exposure in these settings causes around 100 avoidable deaths per year from lung cancer, coronary heart disease and stroke collectively. National survey data and studies in hospitality industry settings (measuring nicotine in hair and cotinine in saliva), strongly indicate that smoke-free workplaces result in reduced exposure to SHS. The data indicate that there is widespread concern regarding SHS and clear support for smoke-free workplaces. CONCLUSIONS: Available New Zealand data are fully consistent with the extensive international data indicating that smoke free policies in workplaces can improve health protection for workers. New legislation is likely to enhance the control of SHS in New Zealand, but additional actions such as a mass media campaign on SHS are also desirable. PMID- 12552287 TI - The Otago University Medical School pre-clinical curriculum: a postgraduate student perspective. PMID- 12552288 TI - Responding to in-flight requests for medical assistance. PMID- 12552289 TI - The monitoring of adverse events following immunization. PMID- 12552290 TI - Regulation of health food stores. PMID- 12552292 TI - Failure of the vaccine cold chain following modification of a domestic refrigerator. PMID- 12552293 TI - Remember to communicate and communicate to remember. PMID- 12552294 TI - Informed consent: patients listen and read, but what information do they retain? AB - AIM: To determine the percentage of knowledge retained immediately following an outpatient consultation for total hip and knee joint arthroplasty, and whether any improvement in that knowledge occurred after reading an information leaflet about the operation. METHODS: Patients on the waiting list for joint replacement surgery were given verbal information during the consultation about basic operative details, post-operative programme, and potential complications. A questionnaire was completed asking them to recall these details. They were then given information leaflets to read. Six weeks later, they were contacted again and asked the same questions. RESULTS: Immediately following a consultation, patients recall only a small percentage of information. In particular, retention of post-operative recovery time frames, and possible operative complications is poor. Despite an information booklet, patients' level of knowledge deteriorates from the initial consultation. CONCLUSIONS: Verbal and written information supplied to a patient may be understood, but it is easily and quickly forgotten. In an increasingly medicolegal environment, it is essential to gain informed consent from a patient when performing interventions. The provision of an information booklet may provide nothing more than proof for the surgeon of information provision to the patient. PMID- 12552295 TI - Diabetes care by general practitioners in South Auckland: changes from 1990 to 1999. AB - AIM: To compare self-reported practices and preferences for diabetes care by general practitioners (GPs) in South Auckland between 1990 and 1999. METHODS: Mail questionnaires were sent to all GPs in South Auckland in 1990, and again in 1999. RESULTS: The response rate was 88% (163/185) in 1990 and 76% (186/245) in 1999 (p = 0.3). In 1999, compared with 1990, GPs had more diabetic patients (median 33 vs 20, p <0.001), more GPs screened for diabetes using a fasting laboratory glucose (33.6% vs 22.9%, p = 0.04), more screened with capillary whole blood testing meters (19.5% vs 1.3%, p <0.001), more felt confident to detect complications (95.1% vs 84.3%, p 0.001) and probably more felt confident to initiate insulin in Type 2 diabetes. Women in both years, compared with male GPs, were more likely to refer newly diagnosed diabetic patients to secondary services (68.0% vs 42.8%, p <0.001) and more likely to prefer 'shared care' for ongoing care (74.7% vs 58.7%, p = 0.007). CONCLUSIONS: There have been large changes in GP diabetes care in South Auckland from 1990 to 1999. GPs in 1999 seem more confident to care for larger numbers of diabetes patients. Significant differences in practice style exist between male and female GPs. PMID- 12552296 TI - Prevalence and correlates of irritable bowel symptoms in a New Zealand birth cohort. AB - AIM: To determine the prevalence and correlates of bowel symptoms and the irritable bowel syndrome (IBS) in a birth cohort of young New Zealanders. METHODS: Participants in the Dunedin Multidisciplinary Health and Development Study at age 26 completed a validated Bowel Disease Questionnaire expressing their experience of clearly defined symptoms over the previous 12 months. RESULTS: 980 participants (499 male, 481 female, comprising 96% of the birth cohort) completed the questionnaire. Sixty four per cent had at least one of the measured symptoms; abdominal pain was reported in 46.5%, chronic constipation in 9.1%, and chronic diarrhoea in 17.1%. A diagnosis of IBS could be made by using two or more of Manning's diagnostic criteria in 18.8%, three or more criteria in 10.3%, and more than three in 3.3%. Symptoms were more than twice as frequent and severe in females than males. CONCLUSIONS: Bowel-related abdominal symptoms, including those required for a diagnosis of IBS, are very common in 26-year-old New Zealanders; the prevalence of these symptoms is very similar to that recorded previously in Europe and the USA. PMID- 12552297 TI - Prescribing for patients aged 65 years and over in New Zealand general practice. AB - AIM: To describe patterns of prescribing in general practice for New Zealanders aged 65 years and over. METHODS: The computerised records of 139 359 consulting patients from 31 general practices from around New Zealand were examined. A subset of 17 497 consulting patients aged 65 years and over was selected and their prescribed medications examined. Utilisation was described in terms of demographic characteristics and health card eligibility. RESULTS: 84.6% of all consulting patients received one or more prescriptions during the study period. Patients were prescribed a mean of 19.7 medication items per annum. Females were prescribed to significantly more times than males for patients aged 79 and under. Community services card (CSC) holders were prescribed to more frequently than patients without a CSC in all age groups. 29.5% of all medication items were from the cardiovascular system Anatomical Therapeutic Chemical (ATC) grouping. CONCLUSIONS: There is a high level of exposure to medication in populations of older people. This is a reflection of older persons' morbidity and also indicates an urgent need to examine the data further for potential drug interactions and side effects. PMID- 12552298 TI - Privacy pitfalls in doctors' disclosures following an operation. PMID- 12552300 TI - Informed consent. PMID- 12552299 TI - Assessment of snorers in primary care. PMID- 12552301 TI - Caring for children who are dying. PMID- 12552302 TI - Inflammation, atherosclerosis and predicting cardiovascular risk. PMID- 12552303 TI - Hyperhomocysteinaemia: time to screen and treat? PMID- 12552304 TI - The need for paediatric palliative care in New Zealand. AB - AIM: To estimate the incidence of life-limiting conditions requiring palliative care in New Zealand children. METHODS: Mortality data was obtained for all children who died aged 0-17 years during the period 1996-1998. Based on the cause of death and hospitalisation data, cases were classified as either 'palliative' or 'not palliative'. A separate analysis was performed to allow comparison with overseas estimates of paediatric palliative care need. RESULTS: Of a total of 2,122 deaths during the study period, 348 cases (16%) were assessed as potentially having required palliative care, giving a rate of 1.14 per 10 000 children per year. 37% of these deaths were due to cancer, 11% cardiac conditions, 24% congenital anomalies, and 28% other conditions. 29% of these children died in hospital. CONCLUSIONS: The findings of this study are consistent with estimates from overseas studies and will be valuable in planning paediatric palliative care services in New Zealand. PMID- 12552305 TI - Burns treatment for children and adults: a study of initial burns first aid and hospital care. AB - AIM: To assess the adequacy of initial burns first aid treatment in the community and its subsequent impact on treatment outcome. METHODS: Four-month prospective study of consecutive burn patients presenting to Middlemore Hospital. Patients were interviewed to determine initial burns first aid treatment (BFAT) and assessed as "adequate" or "inadequate", then compared with subsequent treatment. Inpatient care was wound debridement with/without dressings (DO/DB) or split skin grafting (SSG). RESULTS: 40.5% of total 121 patients received adequate BFAT, 59.5% did not, p = <0.001. 50% Caucasians received adequate BFAT, compared with 25% Maori and 33% Pacific Island people, p = 0.084. 15.7% of adequate BFAT patients received DO/DB and 6.6% had SSG, compared with 23.4% and 19.3% respectively for inadequate BFAT, p = 0.03. Scald burns occurred most frequently, 4% adequate BFAT scald patients required SSG compared with 20% of inadequate BFAT scald patients, p = 0.003. Maori and Pacific Island people were over-represented as inpatients (collectively 34.8%) when compared to Caucasian (24.8%) or other ethnic groups, p = 0.25. 38% of all patients were children under 10 with inadequate BFAT tendency compared with adults, p = 0.067. Hospital stay decreased among adequate BFAT paediatric patients, p = 0.016. CONCLUSIONS: A public education strategy is required to improve BFAT, targeting at-risk communities. Following this, the study should be repeated to determine the effectiveness of the campaign and any resultant change in community behaviour. PMID- 12552306 TI - A general practice minimum data set for New Zealand. AB - AIM: To assess the completeness of primary care data collected by the Royal New Zealand College of General Practitioners' (RNZCGP) Dunedin Research Unit, and assess the feasibility of creating a New Zealand national minimum data set for primary care. METHODS: Patients from 42 practices contributing data to the Dunedin Research Unit made up the study population. A six-month sample of data was evaluated for completeness, and compliance to a minimum data set structure. Rates of recording patient identifiers, sex, ethnicity, community services card status, consultation identifier and date, prescriptions and Read codes were calculated for each practice and registered patient. RESULTS: Patient demography, NHI and community services card status were all well recorded (date of birth 99.3%, sex 98.9%, NHI 94.8%, CSC 100.0%). Read codes and ethnicity were still poorly recorded, although there was wide variation between practices. CONCLUSIONS: The completeness of data collected by the Dunedin Research Unit appears to be improving, although there is wide variation between contributing practices. The capability to create a primary care national minimum data set exists, but this will not become a reality until suitable education programmes and support are supplied for general practitioners and other staff members who record patient information. PMID- 12552307 TI - Childhood gonorrhoea in Auckland. AB - AIM: To review the Auckland experience of childhood gonorrhoea, with particular regard to inter-agency management and medicolegal outcome. METHODS: Retrospective review of medical records at a specialist child abuse assessment unit, and follow up with other agencies involved. RESULTS: Twelve cases of genital gonorrhoea in pre-pubertal children in five years, in contrast to two cases in the preceding six years. Major problems in inter-agency coordination, and an inability to identify the source of the infection in most cases. CONCLUSIONS: Gonorrhoea in the pre-pubertal child is an increasing problem, and is often poorly handled. Proposals for an inter-agency guideline are discussed. PMID- 12552308 TI - A puzzling elevation of troponin T. PMID- 12552309 TI - More about cardiovascular disease and lipid management in New Zealand. PMID- 12552310 TI - Cancer genomics: the chips are on the table. PMID- 12552311 TI - Medical records. PMID- 12552312 TI - The value of Nurse Practitioners. PMID- 12552313 TI - The future of the New Zealand Medical Association. PMID- 12552314 TI - The significance of asymptomatic hyperuricaemia. PMID- 12552315 TI - Amikacin as a replacement aminoglycoside for netilmicin for coagulase-negative staphylococcal sepsis in neonates. PMID- 12552316 TI - Regulation of mitochondrial biogenesis by thyroid hormone. AB - Thyroid hormone (T3) has a profound effect on mitochondrial biogenesis. T3 regulated gene expression is mediated by thyroid hormone receptor (TR) binding to thyroid hormone response elements (TREs). In concert with the action of various coactivators and corepressors this interaction leads to a modulation of the chromatin structure and subsequently to a modulation of gene expression of adjacent target genes. However, as numerous genes are endogenously regulated by T3, and a TRE appears to be absent in their regulatory elements, a TR-independent pathway of T3-mediated gene regulation is likely. In this review, we discuss the direct mechanisms of TR-dependent regulation of gene expression on the nuclear and mitochondrial genome by T3. We also summarise recent observations on an indirect mechanism of T3 action via intermediate factor(s). We discuss the regulation of nuclear respiratory factor 1 (NRF-1) and peroxisome proliferator activated receptor gamma coactivator 1 alpha (PGC-1alpha) by T3, suggesting NRF-1 and PGC-1alpha as attractive candidates for an intermediate factor of T3 action in vivo. PMID- 12552317 TI - Nucleotide and haplotypic diversity of the NOS2A promoter region and its relationship to cerebral malaria. AB - To assess the hypothesis that nitric oxide is critical in the pathogenesis of cerebral malaria, we analysed genetic variation in the proximal promoter region of NOS2A, the gene encoding inducible nitric oxide synthase. Sequencing 72 Gambian chromosomes revealed 11 single nucleotide polymorphisms in 2.5 kB (theta=8.6 x 10(-4)). Genotyping 104 nuclear families identified six common haplotypes. A single haplotype, uniquely defined by the NOS2A-1659T allele, was associated with cerebral malaria by a transmission disequilibrium test of 334 affected children and their parents (P=0.02). An independent case-control study of 505 different children from the same population replicated the allelic association with cerebral malaria (odds ratio: 1.31, P=0.04). Taken together these data indicate a weak but significant association of the NOS2A locus with susceptibility to cerebral malaria. Despite high linkage disequilibrium across the region studied, this association would not have been detected without the initial construction of a dense marker set for haplotype tagging. PMID- 12552318 TI - The novel imprinted carboxypeptidase A4 gene ( CPA4) in the 7q32 imprinting domain. AB - By a search for novel human imprinted genes in the vicinity of the imprinted gene MEST, at chromosome 7q32, we identified the carboxypeptidase A4 gene ( CPA4) in a gene cluster of the carboxypeptidase family, 200 kb centromeric to MEST. Because CPA4 was originally identified as a protein induced in a prostate cancer cell line (PC-3) by histone deacetylase inhibitors, and was located at the putative prostate cancer-aggressiveness locus at 7q32, we investigated its imprinting status in fetal tissues and in adult benign hypertrophic prostate (BPH). RT-PCR using four intragenic polymorphisms as markers showed that CPA4 was expressed preferentially from the maternal allele in the fetal heart, lung, liver, intestine, kidney, adrenal gland, and spleen, but not in the fetal brain. It was also preferentially expressed in the BPH. These findings support that CPA4 is imprinted and may become a strong candidate gene for prostate cancer aggressiveness. As a Silver-Russell syndrome (SRS) locus has been proposed to be located to a region near MEST and to be involved in imprinting, CPA4 would have been a candidate gene for SRS. However, analysis of ten SRS patients revealed no mutations in CPA4. PMID- 12552319 TI - Oxytocin infusion in labor: the effect different indications and the use of different diluents on neonatal bilirubin levels. AB - OBJECTIVE: To investigate the relationship of neonatal bilirubin levels to oxytocin infusion and the diluent used for oxytocin infusion. MATERIALS AND METHODS: The study was carried out as a prospective, randomized study in Istanbul University Cerrahpasa School of Medicine, Department of Obstetrics and Gynecology between January to December in 1995. A total of 80 patients managed with oxytocin during labor, enrolled to the study. These patients randomly divided into isotonic % 0.9 saline (Group 1) and 5% glucose solutions (Group 2) by a consecutive order using a balanced block randomization scheme. Forty multiparous patients delivering without oxytocin infusion formed the control group (Group 3). The details of maternal age, gestational age, labor duration, mode of delivery, birth weight of the babies, total volume of fluid administered until delivery and total oxytocin dose were noted in each case. Sodium and initial bilirubin levels were measured in the cord blood. Later on, capillary blood bilirubin and hematocrit concentrations were measured on day 1 and 2 in the newborn nursery. The groups were compared according to these parameters. RESULTS: The data of 29 patients in Group 1, 36 patients in Group 2 and 40 patients in Group 3 were suitable for analysis. The difference between study and control groups regarding the rate of hyponatremia, neonatal hyperbilirubinemia and neonatal jaundice was not statistically significant. Cord plasma sodium levels, cord plasma bilirubin levels and day 1 and 2 hematocrit and plasma bilirubin levels were not statistically different between the groups. irrespective of the diluent used, the cord plasma bilirubin levels and day 2 plasma bilirubin levels were significantly higher in the accelerated group. CONCLUSION: No significant effect of oxytocin infusion was revealed on neonatal hyperbilirubinemia unless oxytocin was for the augmentation of labor. PMID- 12552320 TI - The prognostic implications of anaemia in the outcome of patients with early stages of uterine cervix carcinoma. AB - We studied the prognostic value of anaemia in the evolution of patients with early stages of uterine cervix cancer and treated with radical surgery. An observational study of 114 patients treated for cervical cancer at the "La Fe" Maternity Hospital in Valencia (Spain) during the period 1971 to 1989. Survival analyses were carried out whereby both recurrence and mortality rates were considered. The level of haemoglobin influences the prognosis of the patients in the study presented, and explains a variation in the disease-free interval in correlation with that of tumour size. However, its influence on the survival interval proved to be somewhat less. Its predictive value is not diminished when associated with other important factors regarding the influence on patient evolution and is seen to be a protector variable against recurrence. Patients with haemoglobin levels of less than 13 gr/dl have a less favourable prognosis and this prognosis worsens still further when levels are lower than 12 gr/dl, which is more frequently the case in patients under 40 years of age and with a greater stromal invasion depth. The influence of haemoglobin levels is equally as important in its influence on prognosis and patient evolution as the volume of the tumour itself. The effect of this variable depends on both the clinical characteristics of the patients and the pathological characteristics of the tumour. PMID- 12552321 TI - Anxiety scores before and after prenatal testing for congenital anomalies. AB - OBJECTIVE: Prenatal diagnosis has psychological effects on the pregnant woman. We studied the state of anxiety levels in 332 pregnant women presenting between the 12th and 20th week of gestation before and after prenatal examination with negative and positive prenatal findings. RESULTS: Prenatal examination produced a positive finding in 37 cases (11.1%). Both before and after prenatal examination pregnant women with a positive finding showed significantly higher anxiety-scales when compared to women with a negative result. After the examination state anxiety levels were significantly reduced with a negative prenatal scan, whereas anxiety levels remained constant with a positive result. CONCLUSIONS: Prenatal testing for fetal anomalies presents an anxiety-inducing situation for the parents-to-be. If the results are negative, anxiety is reduced. Pregnant women who are confronted with a positive malformation scan show unchanged levels of anxiety after the examination. PMID- 12552322 TI - The effect of epidural analgesia on obstetric lacerations and neonatal outcome during spontaneous vaginal delivery. AB - The aim of this study was to determine if epidural analgesia is associated with increased risk of obstetric lacerations during spontaneous vaginal delivery. We also assessed the effect of epidural analgesia on maternal and neonatal parameters. This multicenter study consisted of an analysis of data from the delivery databases of the University Hospital of Vienna and the Semmelweis Women's Hospital Vienna. This study was restricted to a sample that included all women with uncomplicated pregnancy, a gestational age >37 weeks and a pregnancy with cephalic presentation. Epidural analgesia was started during the first stage of labour. Techniques and management styles of epidural analgesia were the same in both hospitals. We found that women undergoing epidural analgesia had a prolonged second stage of labour, a higher rate of episiotomy and an increased use of oxytocin. Some of these adverse effects might be caused by the higher rate of primipara in the epidural group. However, epidural analgesia showed no evidence of a detrimental effect on the integrity of the birth-canal and on neonatal outcome during spontaneous vaginal delivery. PMID- 12552323 TI - The effect of an acute decrease in serum estrogen concentration on vessel walls: determination with color and pulsed Doppler ultrasound. AB - The aim of this study was to determine the effects of an acute decrease in serum estrogen concentration on endothelial function in women with surgically induced menopause through the use of color Doppler ultrasonography. There were 40 women scheduled to undergo total abdominal hysterectomy and bilateral salpingo oopherectomy who participated in the study; 15 women not undergoing surgery also participated as a control group. Color Doppler ultrasonographic examinations of each surgical patient were obtained 3 days prior to and 7 days after surgery. Baseline measurements of the brachial arteries, including peak systolic velocity, end-diastolic velocity, true mean velocity, arterial diameter, and volume flow, were obtained for each patient. After baseline measurements were established, hyperemia was induced by inflating a blood pressure cuff on each patient's upper arm to suprasystolic pressures for 5 min. To evaluate endothelium-dependent vasodilation, the ultrasonographic appearance of the brachial artery was evaluated after the cuff was deflated and removed from the arm. Measurements of peak systolic velocity, end-diastolic velocity, true mean velocity, arterial diameter, and volume flow were obtained, and were repeated at 1, 3, 5, 10, and 20 min subsequent to removal of the blood pressure cuff. The differences between baseline and maximum values of each Doppler parameter after the cuff deflation were calculated. No significant differences were identified in terms of laboratory findings or systolic and diastolic pressures in pre- and postoperative status of surgical patients, or between surgical patients and the control group. A significant difference in serum estradiol levels during pre- and postoperative periods ( P<0.001) was detected. No significant difference in serum estradiol levels was detected among preoperative surgical patients and members of the control group ( P=0.72). All net changes detected within each group during reactive hyperemia were statistically significant. No significant difference in values was detected among pre-, postoperative, and control subjects. Our study reveals that acute decrease in serum estrogen level does not appear to affect endothelial function; thus, we assume that this is mainly due to the result of postoperative surgical stress. PMID- 12552324 TI - Effects of tibolone on human breast cancer cells and human vascular coronary cells. AB - Tibolone is a synthetic progestin with estrogenic and progestogenic properties, used to alleviate menopausal syndromes and for osteoporosis prophylaxis in postmenopausal women. However, only little data are available on tibolone and breast cancer risk and on the effects of tibolone on the cardiovascular system. Therefore, in the present in vitro study, we investigated the effect of tibolone on the growth of the human breast cancer cell line, MCF-7, and on the direct effects of tibolone on the vasculature, i.e. human female coronary endothelial and smooth muscle cells. In the breast cancer cell experiments, tibolone was examined alone and in the presence of 0.1 nM estradiol in the concentration range from 0.001 microM to 1 microM. Tibolone lead to significant cell growth in the concentration range of 0.01 micro M to 1 microM and was not able to inhibit estradiol-induced proliferation at the concentrations of 0.01 microM and 0.1 microM. In the vascular endothelial cell culture experiments, tibolone was tested at the concentrations 0.1 microM, 1 microM and 10 microM. Tibolone reduced the synthesis of endothelin as well as the concentrations of E-selectin, PAI-1 and pro-MMP-1. The magnitude of the effects on these markers varied and was in the range of 11%-42%. Concerning smooth muscle cells, tibolone elicited no changes in the proliferation compared to control values. These data suggest that tibolone does have tumour cell-growth promoting effects in vitro. However, tibolone can positively influence the synthesis of markers in cell cultures of human female coronary artery, which modulate vascular tone and which play a decisive role in the various stages of atherosclerosis. Drawing a clinical consequence from our experiments would result in not recommending the use of tibolone in postmenopausal women at high risk for breast cancer development until long-term controlled clinical studies have been performed on the effect of tibolone administration and breast cancer risk. Experimental studies, such as the present one, are useful to explore mechanisms, but clearly cannot replace clinical studies. PMID- 12552325 TI - Factors affecting the survival of 121 patients treated for endometrial carcinoma at a Polish hospital. AB - A retrospective analysis was made of 121 patients treated for endometrial carcinoma. Patients who were married and were professionally active at the time of the diagnosis of endometrial cancer had a significantly longer predictive 5 year survival in comparison to other groups of patients. A significantly longer expected 5-year survival was observed in married patients compared to widowed and single patients (P=0.01). Patients who were professionally active at the time of diagnosis had a longer expected 5-year survival than retired patients and housewives (P=0.05). Statistically better prognoses were confirmed in patients with clinically less advanced disease (P=0.003) and a lesser degree of histologic differentiation (P=0.04). However, obese patients had a longer 5-year survival compared to non-obese patients and the difference was statistically significant (P=0.05). Body-mass index (BMI) in the range of 25-29 had a favourable influence on predictive 5-year survival (P=0.05). PMID- 12552326 TI - Incidence of gestational hypertension in gestational diabetes mellitus. AB - The purpose of this study has been to establish the incidence of gestational hypertension (GH) in women with gestational diabetes mellitus (GDM) and to examine the frequency of complications in women with co-existent GDM and GH. Furthermore, we wished to evaluate the significance of urine albumin excretion determined by the urine albumin creatinine ratio (ACR). A total of 215 successive pregnancies with risk factors for gestational diabetes, as defined by the Danish National Board of Health screened for gestational diabetes by the WHO criteria, were reviewed. Women who had a normal OGTT during the screening served as controls. Gestational hypertension was defined as a mean arterial pressure (MAP) >105 mmHg (systolic BP > or = 140 mmHg and/or diastolic BP > or = 90 mmHg). The two groups were comparable with regard to initial body mass index (BMI) and MAP. GH appeared with a higher frequency in women with GDM (28%) than in women with normal OGTT (10%) P=0.003 chi(2) test. Serious complications (perinatal mortality, malformations, acute caesarean section) also appeared with a higher frequency in women with GH and GDM (10%) than in women with GH but with normal OGTT (2%) P=0.0083 chi(2) test. We observed a significant increase in ACR in the group with complications (GDM and GH) during gestation regardless of intensive antihypertensive treatment. We also observed that ACR was significantly higher in women with GDM and GH when compared to women with GDM and a normal blood pressure. The BMI was consistently higher in women with GH, regardless of whether they had GDM or not as compared to the normotensive group. GH appears with a higher frequency in women with GDM and the co-existence seems correlated with a higher frequency of complications. The correlation between urine albumin excretion and complications might suggest that regulating GH should strive to normalise ACR in women with GDM. PMID- 12552327 TI - Giant mucinous cystadenocarcinoma: a case report. AB - We present a mucinous cystadenocarcinoma weighing 10 kg in a postmenopausal woman and the way how we managed the case. PMID- 12552328 TI - An unusual case of postmenopausal vaginal bleeding: retention of fetal bone. AB - Retention of intrauterine fetal bones is a rare finding in patient suffering from abnormal uterine bleeding or secondary infertility. Detailed patient history, pelvic ultrasonography and hysteroscopy are diagnostic tools. Here, we describe a case of postmenopausal persistent uterine bleeding and pelvic pain caused by prolonged retention of fetal bones after a midtrimester abortion 17 years ago. PMID- 12552329 TI - Fatal maternal outcome of a parturient with Eisenmenger's syndrome and severe preeclampsia. AB - Eisenmenger's syndrome in pregnancy is associated with a high maternal and fetal morbidity and mortality. When it occurs with severe preeclampsia, the morbidity and mortality are higher. We report the case of a 30 weeks' pregnant woman with Eisenmenger's syndrome and severe preeclampsia. Cesarean section was performed due to severe preeclampsia and an unfavorable cervix under general anesthesia. The intraoperative period was uneventful and a healthy 1300 g male infant was delivered, but the patient died on the second postoperative day due to a pulmonary embolism. This case confirms the frequently fatal maternal outcome of Eisenmenger's syndrome in pregnancy. Early termination of pregnancy is the treatment of choice. PMID- 12552330 TI - Ossifying luteinized thecoma of the ovary. AB - A 51-year-old woman presented with lumbar backache leading to the preoperative diagnosis of a right solid adnexal mass with calcification on computed tomographic scan. Histological examination revealed a right ovarian luteinized thecoma characterized by extensive calcification and metaplastic ossification. Osteoblasts and osteoclasts surrounded the surface of the heterotopic bone. Haversian canals were occasionally identified in the bony trabeculae. Ossifying ovarian neoplasms are extremely rare and this case is the first to demonstrate the osseous metaplasia in ovarian luteinized thecoma. PMID- 12552331 TI - Ruptured interstitial pregnancy Two case reports. AB - The authors described two cases of the ruptured unrecognized interstitial pregnancy. 32-year-old woman, was admitted for abdominal colics in 17th week of uncontrolled pregnancy. Obstetric finding on admission was normal, while ultrasonography pointed to 17th week of gestation. During the first night of hospitalization, severe abdominal pain, with collapse, tachycardia and hypotension developed. Laparotomy was performed in general endotracheal anesthesia with resuscitation measures, and revealed hematoperitoneum and ruptured right-sided interstitial pregnancy with a dead fetus among the bowels. Abdominal hysterectomy without adnexa was performed. S. J., a 35-year-old tertigravida, six years before, left-sided adnexectomy was performed for perforated tubal pregnancy. 5 weeks before artificial abortion now, ultrasonography revealed a vital fetus biometrically corresponding to the 11th week of gestation. The condition was recognized as an unsuccessful artificial abortion and intact pregnancy. During preoperative preparation the patient lost consciousness and required immediate resuscitation. The abdominal cavity was opened by Pfannenstiel's relaparotomy, and revealed hematoperitoneum with numerous adhesions between the small intestine, sigma, uterus and urinary bladder, and ruptured left-sided interstitial pregnancy. Adhesiolysis was completed, a lesion in the sigma was sutured, and hysterectomy was performed. Exploration of the abdominal cavity revealed a dead fetus among small intestine loops, and an indurated, sharply delineated neoplasia of the head of the pancreas. PMID- 12552332 TI - Gonadal dysgenesis and the Mayer-Rokitansky-Kuster-Hauser syndrome in a girl with 46,X,del(X)(pter-->q22:). AB - The association of mullerien duct with gonadal dysgenesis is extremely rare. We report such a case in a 19 year-old white woman with a 46,X,del(X)(pter-->q22:) karyotype. PMID- 12552333 TI - Ruptured ovarian granulosa cell tumors as a cause of the acute abdomen. AB - A rare case of acute abdomen syndrome due to a ruptured ovarian tumor and a penetrated fallopian tube is described. Based on the intraoperative finding and patient's age, a right-sided salpingo-oophorectomy, appendectomy and peritoneal washings were performed. After a histological diagnosis of malignant granulosa cell tumors and FIGO IIA clinical classification, the patient returned 1 month after the procedure. A relaparotomy with a hysterectomy, left-sided salpingo oophorectomy and omentectomy were then performed. No spread was found during the laparotomy, and the histologic diagnosis pointed to adenomyosis and chronic cervicitis. The patient regularly presents for control examination, and has now been free for 11 years since the surgery from clinical, biochemical or ultrasonographic signs of a relapse of the disease. PMID- 12552334 TI - Quantitative cerebral H2(15)O perfusion PET without arterial blood sampling, a method based on washout rate. AB - The quantitative determination of regional cerebral blood flow (rCBF) is important in certain clinical and research applications. The disadvantage of most quantitative methods using H(2)(15)O positron emission tomography (PET) is the need for arterial blood sampling. In this study a new non-invasive method for rCBF quantification was evaluated. The method is based on the washout rate of H(2)(15)O following intravenous injection. All results were obtained with Alpert's method, which yields maps of the washin parameter K(1) (rCBF(K1)) and the washout parameter k(2) (rCBF(k2)). Maps of rCBF(K1) were computed with measured arterial input curves. Maps of rCBF(k2*) were calculated with a standard input curve which was the mean of eight individual input curves. The mean of grey matter rCBF(k2*) (CBF(k2*)) was then compared with the mean of rCBF(K1) (CBF(K1)) in ten healthy volunteer smokers who underwent two PET sessions on day 1 and day 3. Each session consisted of three serial H(2)(15)O scans. Reproducibility was analysed using the rCBF difference scan 3-scan 2 in each session. The perfusion reserve (PR = rCBF(acetazolamide)-rCBF(baseline)) following acetazolamide challenge was calculated with rCBF(k2*) (PR(k2*)) and rCBF(K1) (PR(K1)) in ten patients with cerebrovascular disease. The difference CBF(k2*)-CBF(K1) was 5.90+/ 8.12 ml/min/100 ml (mean+/-SD, n=55). The SD of the scan 3-scan 1 difference was 6.1% for rCBF(k2*) and rCBF(K1), demonstrating a high reproducibility. Perfusion reserve values determined with rCBF(K1) and rCBF(k2*) were in high agreement (difference PR(k2*)-PR(K1)=-6.5+/-10.4%, PR expressed in percentage increase from baseline). In conclusion, a new non-invasive method for the quantitative determination of rCBF is presented. The method is in good agreement with Alpert's original method and the reproducibility is high. It does not require arterial blood sampling, yields quantitative voxel-by-voxel maps of rCBF, and is computationally efficient and easy to implement. PMID- 12552335 TI - 99mTc-tetrofosmin or 99mTc-sestamibi for double-phase parathyroid scintigraphy? AB - Several years ago technetium-99m tetrofosmin was reported to localise parathyroid adenomas. The aim of this study was to compare the sensitivity of this radiopharmaceutical with that of (99m)Tc-sestamibi using a double-phase parathyroid scintigraphy protocol. Scans of 12 patients were evaluated visually and lesion to thyroid ratios were calculated. Nine of the patients were subsequently operated on; a total of eight parathyroid adenomas or hyperplastic glands were histologically confirmed in seven of the patients, while in one patient a parathyroid carcinoma was histologically proven. All of these patients had positive (99m)Tc-sestamibi scintigrams, whereas only two (99m)Tc-tetrofosmin scintigrams were positive. With (99m)Tc-sestamibi there was a significant increase in the lesion to thyroid ratio from 10 min to 90 min and 150 min p.i. which was not seen on scintigraphy with (99m)Tc-tetrofosmin. This makes (99m)Tc tetrofosmin less suitable for double-phase parathyroid scintigraphy. PMID- 12552336 TI - Diagnostic and prognostic value of 99mTc-tetrofosmin scintigraphy in maxillofacial flaps. AB - In oro-maxillofacial malignancies, new therapeutic approaches are placing changing demands on the diverse diagnostic modalities. In contradistinction to mandibular reconstruction of former years, the transplants (microvascular anastomosed pedicled flaps, "flaps") now consist of one or more arteries feeding a soft tissue component attached to a piece of bone suitably fitted to fill the defect. We addressed the diagnostic value of technetium-99m tetrofosmin scintigraphy in differentiating between viability and non-viability of the soft tissue portion of flaps in the immediate postoperative assessment. A total of 60 patients who had received flaps for reconstruction of the mandible after partial resection were investigated with (99m)Tc-tetrofosmin 3-5 days after surgery. Scintigraphy consisted of (a) radionuclide angiography, (b) static planar imaging in four projections starting at 10 min post injection, and (c) single-photon emission tomography (SPET) performed immediately after the planar imaging. Normal perfusion associated with no defects throughout the soft tissue portion of the transplant was observed in 46/60 patients. This scintigraphic pattern was identical to viability and normal postoperative follow-up. Hypoperfusion and small defects on planar and SPET images indicated viability and uncomplicated postoperative healing in 6/60 patients, but non-viability/inadequate healing of the flap in 4/60 patients. Absence of perfusion combined with a large defect on static planar and SPET images definitively showed the non-viability of the flap (4/60 patients). It is concluded that (99m)Tc-tetrofosmin scintigraphy is a sensitive diagnostic tool for the immediate postoperative assessment of the viability and the adequacy of implantation of the soft tissue portion of flaps. Therefore tetrofosmin scintigraphy is an important modality in order (a) to define the optimal therapeutic regimen in the immediate postoperative period and (b) to provide better prognosis. PMID- 12552337 TI - Dynamic lymph flow imaging in patients with oedema of the lower limb for evaluation of the functional outcome after autologous lymph vessel transplantation: an 8-year follow-up study. AB - The purpose of this study was to monitor the functional outcome of microsurgical intervention on lymph drainage by means of non-invasive, readily available lymphoscintigraphy. Eight patients with primary or secondary lymphoedema of the lower limb were investigated before and for 8 years after autologous lymph vessel transplantation. For scintigraphy, technetium-99m labelled nanocolloid was subcutaneously injected into the first interdigital space of the affected limb. Sequential images were acquired up to 6 h p.i.; for semiquantitative evaluation a numerical transport index was established by assigning scores of up to 9 on each of five criteria: lymphatic transport kinetics, distribution pattern of the radiopharmaceutical, time to appearance of lymph nodes, visualisation of lymph nodes and visualisation of lymph vessels/grafts. Ti values <10 were considered normal. In all eight patients, lymphatic function significantly (P/=99.5%; however, since a total patient dose can be as high as 800 mCi, the amount of free (177)Lu(3+) (= non-DOTA-incorporated) can be substantial. Free (177)Lu(3+) accumulates in bone with unwanted irradiation of bone marrow as a consequence. (177)Lu-DTPA is reported to be stable in serum in vitro, and in vivo it has rapid renal excretion. Transforming free Lu(3+) to Lu-DTPA might reroute this fraction from accumulation in bone to renal clearance. We therefore investigated: (a) the biodistribution in rats of (177)LuCl(3), [(177)Lu-DOTA(0),Tyr(3)]octreotate and (177)Lu-DTPA; (b) the possibilities of complexing the free (177)Lu(3+) in [(177)Lu-DOTA(0),Tyr(3)]octreotate to (177)Lu-DTPA prior to intravenous injection; and (c) the effects of free (177)Lu(3+) in [(177)Lu DOTA(0),Tyr(3)]octreotate, in the presence and absence of DTPA, on the biodistribution in rats. (177)LuCl(3) had high skeletal uptake (i.e. 5% ID per gram femur, with localization mainly in the epiphyseal plates) and a 24-h total body retention of 80% injected dose (ID). [(177)Lu-DOTA(0),Tyr(3)]octreotate had high and specific uptake in somatostatin receptor-positive tissues, and 24-h total body retention of 19% ID. (177)Lu-DTPA had rapid renal clearance, and 24-h total body retention of 4% ID. Free (177)Lu(3+) in [(177)Lu DOTA(0),Tyr(3)]octreotate could be complexed to (177)Lu-DTPA. Accumulation of (177)Lu in femur, blood, liver and spleen showed a dose relation to the amount of free (177)Lu(3+), while these accumulations could be normalized by the addition of DTPA. After labelling [DOTA(0),Tyr(3)]octreotate with (177)Lu the addition of DTPA prior to intravenous administration of [(177)Lu-DOTA(0),Tyr(3)]octreotate is strongly recommended. PMID- 12552353 TI - Improved detection of splenosis in patients with haematological disorders: the role of combined transmission-emission tomography. AB - The purpose of this study was to evaluate the use of combined transmission and emission tomography (TET) for correct localisation of heterotopic splenic tissue and differentiation of splenosis from other masses. The TET technique comprises the fusion of SPET and CT data obtained using the same imaging device to allow perfect overlap of anatomical and functional images. TET was performed in seven patients who either had haematological disorders and relapsing anaemia or thrombocytopenia after splenectomy or were under immunosuppression for different reasons. These patients presented 20 equivocal lesions on CT or MRI. Presence of splenic tissue was investigated using technetium-99m labelled colloids or heat damaged red blood cells. Findings of spleen scintigraphy, TET and CT or MRI were compared with respect to localisation of splenosis and correct classification of lesions by CT or MRI. Histological validation was achieved by surgery or biopsy in all cases. All 20 lesions demonstrated by CT or MRI were correctly classified by TET as splenosis. Three additional lesions initially overlooked by CT or MRI could be detected. Diagnostic relevance was highest for intrahepatic, intrapulmonary or pleural splenic implants. It is concluded that TET allows exact localisation of heterotopic splenic tissue in patients with suspected splenosis. PMID- 12552354 TI - Activity and practice of nuclear cardiology in the Czech Republic, 2001. AB - Radionuclide myocardial perfusion imaging (MPI) has been on the rise in Europe and the USA. Details on nuclear cardiology in the Czech Republic are not available as yet, as it is impossible to obtain comprehensive data from official registers owing to different methods of reporting and data evaluation. A questionnaire concerning nuclear cardiology activity and practice in 2001 was sent to all nuclear medicine departments in the Czech Republic. All 48 departments completed the questionnaire. In 2001, 50 planar and 54 tomographic (SPET) scintillation cameras were used. The average age of the SPET cameras was 5 years (13% of SPET cameras were >8 years old). Out of the 48 centres, 39 (81%) provided a nuclear cardiology service; the total number of cardiological studies was 15,740 in 2001 (1.5 studies/1,000 population/year). The most frequently employed method was MPI (81.7%), the frequency of which had increased by 10% compared with 2000; 26 of the 39 (67%) departments reported that MPI activity was increasing. Nevertheless, the Czech Republic nuclear cardiology activity remained below the European average (2.2/1,000 population in 1994) and, particularly, below activity in the USA (15/1,000 in 1997). The activity was rather unevenly spread. Whereas two centres with >1,000 studies/year accounted for 20% of the total MPI studies, 16 of 39 (41%) departments exhibited low activity (<200 studies/year) and accounted for only 15% of the total MPI studies. The use of SPET increased from 91% in 2000 to 94% in 2001 (only three institutes performed planar examinations). The most widely used tracer was (99m)Tc-MIBI (60% of total MPI), followed by (201)Tl (21%) and (99m)Tc-tetrofosmin (19%). ECG-gated SPET was employed by 20/39 (51%) centres, of which 11 (28%) performed it as a standard examination; 39% of the total MPI studies included this technique. Thirteen percent (5/39) of the departments used attenuation correction, and 69% (27/39) of the departments used a prone projection. Equilibrium radionuclide ventriculography, with 2,317 examinations (14.7%), ranked second among all nuclear cardiology methods, followed by first-pass angiocardiography (406 studies, 2.6%) and (18)F-FDG (163 studies, 1%). PMID- 12552355 TI - Is central benzodiazepine receptor imaging useful for the identification of epileptogenic foci in localization-related epilepsies? AB - In the presurgical evaluation of patients with partial epilepsies, the most extensively studied functional neuro-imaging modality to define the origin of seizure onset is fluorine-18 fluorodeoxyglucose positron emission tomography (FDG PET). Generally, this technique reveals a widespread zone of interictal glucose hypometabolism in the region of the epileptogenic focus. However, the technique may miss the epileptogenic region and FDG PET abnormalities may extend beyond the seizure onset zone. Consequently, for the precise identification of epileptogenic regions more specific imaging probes than FDG are warranted. This review considers the clinical utility of iomazenil (IMZ) SPET and flumazenil (FMZ) PET for the precise localization of epileptogenic foci in partial epilepsy syndromes. PMID- 12552356 TI - Enhancement of uptake of therapeutic radiopharmaceuticals in bone lesions. PMID- 12552357 TI - Hypertrophic osteoarthropathy on bone scintigraphy related to a mediastinal tumour of unknown origin imaged by FDG-PET. PMID- 12552358 TI - The 5-HT1A agonist 8-OH-DPAT dose-dependently interferes with the establishment and the expression of lithium-induced conditioned rejection reactions in rats. AB - RATIONALE: The present experiments evaluated the potential of the 5-HT(1A )agonist, 8-OH-DPAT (DPAT), which reduces serotonin availability, to interfere with both the establishment and with the expression of lithium-induced conditioned rejection reactions (experiment 1) and lithium-induced taste avoidance (experiment 2). OBJECTIVES. To determine the effect of reduced serotonin availability on conditioned rejection reactions, a rat model of nausea. METHODS: Rats were injected with 8-OH-DPAT [at doses of 0.0 (saline), 0.01 or 0.1 mg/kg, SC] 30 min prior to exposure to 0.1% saccharin solution by intra-oral infusion (experiment 1) or by bottle presentation (experiment 2). Immediately following saccharin exposure, rats were injected with 20 ml/kg lithium chloride (0.15 M) or 20 ml/kg saline solution. On each of three test trials, rats were injected with DPAT [0.0 (saline), 0.01 and 0.1 mg/kg, SC; counterbalanced order], 30 min prior to exposure to saccharin solution by intra-oral infusion (experiment 1) or by a two-bottle test (experiment 2: saccharin and water). RESULTS: DPAT interfered with both the establishment (at a dose of 0.1 mg/kg, SC) and with the expression (at doses of 0.01 and 0.1 mg/kg, SC) of lithium-induced conditioned rejection reactions; however, DPAT did not modulate taste avoidance in a consumption test. CONCLUSIONS: These results indicate that conditioned rejection reactions, but not taste avoidance, can be attenuated by the anti-emetic agent, 8 OH-DPAT. PMID- 12552359 TI - Distinctive effects of modafinil and d-amphetamine on the homeostatic and circadian modulation of the human waking EEG. AB - RATIONALE: Modafinil is a wake-promoting agent that affects hypothalamic structures involved in the homeostatic and circadian regulation of vigilance. Administered during sleep deprivation, it reduces the need for prolonged recovery sleep and decreases the rebound in EEG slow-wave activity. These diachronic effects suggest an action of modafinil on a homeostatic sleep regulatory process. OBJECTIVES: The aim of this study was to determine whether modafinil, in comparison to the d-amphetamine reference psychostimulant and to placebo, interferes with the vigilance regulatory processes reflected in the EEG during waking. METHODS: Thirty-three healthy subjects were investigated during 60 h of sustained wakefulness in a double-blind placebo-controlled parallel-design study. A 4-min maintenance-of-wakefulness test administered hourly allowed the concomitant assessment of alertness and waking EEG activity. The effects of equipotent psychostimulant dosages (modafinil 300 mg and d-amphetamine 20 mg) were evaluated at the beginning of the first sleep deprivation night, at the end of the second sleep deprivation night and in the afternoon preceding the first recovery night. RESULTS: One hour following ingestion, both psychostimulants increased alertness during 10-12 h, independently of the time of administration. At the level of the waking EEG, d-amphetamine attenuated the natural circadian rhythm of the different frequency bands and suppressed the sleep deprivation related increase in low frequency (0.5-7 Hz) powers. In contrast, modafinil, which exhibited a transient amphetamine-like effect, had slight effect on circadian rhythms. Its selective action was characterized by maintenance of the alpha(1) (8.5-11.5 Hz) EEG power, which under placebo exhibited a homeostatic decrease paralleling that of alertness with a circadian trough at night. CONCLUSIONS: These findings demonstrate that the alertness-promoting effects of modafinil and d-amphetamine involve distinct EEG activities and do not reside on the same vigilance regulatory processes. While d-amphetamine inhibits the expression of a sleep-related process, probably through a direct cortical activation masking EEG circadian rhythms, modafinil, through a synchronic effect, preferentially disrupts the homeostatic down-regulation of a waking drive. PMID- 12552360 TI - Buspirone-induced antinociception is mediated by L-type calcium channels and calcium/caffeine-sensitive pools in mice. AB - RATIONALE: Previous studies have shown that buspirone, a partial 5-HT(1A) receptor agonist, produces antinociceptive effects in rats and mice; Ca(2+) plays a critical role as a second messenger in mediating nociceptive transmission. 5 HT(1A) receptors have been proven to be coupled functionally with various types of Ca(2+) channels in neurons, including N-, P/Q-, T-, or L-type. It was of interest to investigate the involvement of extracellular/intracellular Ca(2+) in buspirone-induced antinociception. OBJECTIVES: To determine whether central serotonergic pathways participate in the antinociceptive processes of buspirone, and investigate the involvement of Ca(2+) mechanisms, particularly L-voltage gated Ca(2+) channels and Ca(2+)/caffeine-sensitive pools, in buspirone-induced antinociception. METHODS: Antinociception was assessed using the hot-plate test (55 degrees C, hind-paw licking latency) in mice treated with either buspirone (1.25-20 mg/kg i.p.) alone or the combination of buspirone and fluoxetine (2.5-10 mg/kg i.p.), 5-HTP (25 mg/kg i.p.), nimodipine (2.5-10 mg/kg i.p.), nifedipine (2.5-10 mg/kg i.p.), CaCl(2) (25-200 nmol per mouse i.c.v.), EGTA (5-30 nmol per mouse i.c.v.), or ryanodine (0.25-2 nmol per mouse i.c.v.). RESULTS: Buspirone dose dependently increased the licking latency in the hot-plate test in mice. This effect of buspirone was enhanced by fluoxetine, 5-HTP, nimodipine, and nifedipine. Interestingly, central administration of Ca(2+) reversed the antinociceptive effects of buspirone. In contrast to these, ryanodine or EGTA administered centrally potentiated buspirone-induced antinociception. CONCLUSIONS: Decreasing neuronal Ca(2+) levels potentiated buspirone-induced antinociception; conversely, increasing intracellular Ca(2+) abolished the antinociceptive effects of buspirone. These results suggest that Ca(2+) influx from extracellular fluid and release of Ca(2+) from Ca(2+)/caffeine-sensitive microsomal pools may be involved in buspirone-induced antinociception. PMID- 12552361 TI - Increased extracellular serotonin level in rat hippocampus induced by chronic citalopram is augmented by subchronic lithium: neurochemical and behavioural studies in the rat. AB - RATIONALE: A substantial number of patients do not respond sufficiently to antidepressant drugs and are therefore often co-medicated with lithium as an augmentation strategy. However, the neurochemical rationale behind this strategy needs to be further clarified. OBJECTIVES: We examined the effect of chronic citalopram and subchronic lithium, alone or in combination, on (a) serum levels of citalopram and lithium, (b) animal behaviour and (c) hippocampal serotonin (5 HT) and 5-hydroxyindoleacetic acid (5-HIAA) levels. Furthermore, we examined the serum level of citalopram and hippocampal 5-HT following one acute citalopram injection. METHODS: Microdialysis in the freely moving animals was used to determine hippocampal 5-HT and 5-HIAA. The animal behaviour was examined in the open field and forced swim test. RESULTS. We found that chronic administration of citalopram (20 mg/kg/24 h s.c.) significantly increased the 5-HT baseline relative to vehicle-treated rats. Addition of subchronic lithium (60 mmol/kg chow pellet p.o.) to chronic citalopram therapy further elevated the 5-HT levels. Moreover, we found acute citalopram (5 mg/kg s.c.) to increase the 5-HT level. The immobility time in the FST and the locomotion in the OF were unaffected by any treatments. CONCLUSIONS: The present results support the assumption that increases in hippocampal 5-HT neurotransmission may be important in the augmentatory effect of lithium. PMID- 12552362 TI - Attenuation of hypothalamic-pituitary-adrenocortical hyperactivity in depressed patients by mirtazapine. AB - RATIONALE: It has been suggested that hypothalamic-pituitary-adrenocortical (HPA) system dysregulation plays an important role in the pathophysiology of depression and that normalization of HPA axis hyperactivity precedes successful treatment with antidepressants. Mirtazapine acts as an antagonist at presynaptic alpha(2) receptors and at postsynaptic 5-hydroxytryptamine (5-HT)(2), 5-HT(3) and histamine H(1) receptors. It has been shown acutely to inhibit cortisol secretion in healthy subjects. OBJECTIVE: In this study, we investigated whether mirtazapine may downtune HPA axis hyperactivity in depressed patients and whether this is related to treatment outcome. METHODS: Forty patients suffering from a major depressive episode (DSM-IV criteria) were treated with mirtazapine for 5 weeks. The combined dexamethasone suppression/CRH stimulation test (DEX/CRH test) was performed before and after 1 week of mirtazapine treatment (45 mg daily). RESULTS: Mirtazapine effectively reduced the overshoot of cortisol and ACTH during the DEX/CRH test both in treatment responders and non-responders within 1 week. CONCLUSIONS: Apparently, mirtazapine rapidly attenuates HPA axis hyperactivity in depressed patients via direct pharmacoendocrinological effects. However, this amelioration of HPA system dysregulation is not necessarily related to clinical improvement. PMID- 12552363 TI - Cognitive and motor function after administration of hydrocodone bitartrate plus ibuprofen, ibuprofen alone, or placebo in healthy subjects with exercise-induced muscle damage: a randomized, repeated-dose, placebo-controlled study. AB - RATIONALE: Medications combining hydrocodone bitartrate and non-steroidal anti inflammatory agents appear more beneficial than anti-inflammatory medications alone in treating pain and inflammation from acute soft tissue trauma, but opiate side effects may include sedation and impaired cognitive and motor performance. OBJECTIVE: Performance on complex cognitive and motor tasks was evaluated in healthy subjects with exercise-induced muscle damage who were treated with a hydrocodone-ibuprofen combination, ibuprofen alone, or placebo. METHODS: This double-blind, randomized, placebo-controlled, repeated-dose clinical trial compared the effects of hydrocodone bitartrate (7.5 mg) plus ibuprofen (200 mg), ibuprofen alone, and placebo on cognitive and motor function in 72 healthy college men. Muscle damage in the quadriceps of each subject's dominant leg was induced by an eccentric exercise protocol. Subjects took the study medication four times daily (every 4-6 h) for 5 days. Forty minutes after medication ingestion at the same time each day, subjects underwent tests of attention/concentration, motor performance, and reaction time. Four trained assessors rotated among subjects so that none tested the same participant on more than three occasions. RESULTS: Repeated measures analyses of covariance revealed no between-group differences on a complex memory and cognition task or complex reaction time. Subjects using hydrocodone bitartrate plus ibuprofen performed significantly less well on a simple tracking task and made significantly more errors on a simple reaction-time task than the other two groups. These deficits were found to be highly transitory and not related to confusion or fatigue. CONCLUSION: Hydrocodone plus ibuprofen was not associated with deterioration in complex cognition but was related to very transitory decrements in tasks involving simple hand-eye coordination. PMID- 12552365 TI - Multiple sclerosis and Type I diabetes. PMID- 12552364 TI - High glucose impairs superoxide production from isolated blood neutrophils. AB - OBJECTIVE: Superoxide (O(2)(-)), a key antimicrobial agent in phagocytes, is produced by the activity of NADPH oxidase. High glucose concentrations may, however, impair the production of O(2)(-) through inhibition of glucose-6 phosphate dehydrogenase (G6PD), which catalyzes the formation of NADPH. This study measured the acute effects of high glucose or the G6PD inhibitor dehydroepiandrosterone (DHEA) on the production of O(2)(-) from isolated human neutrophils. DESIGN: Laboratory studies of short-term cultures of neutrophil granulocytes. PARTICIPANTS: Healthy subjects. INTERVENTIONS: Neutrophils were isolated from peripheral blood and incubated for 1 h in Krebs-Ringer buffer containing 5, 10, or 25 mM glucose, 5 mM glucose with 0, 5, or 20 mM mannitol, or 5 mM glucose with 0, 1, 10, or 100 micro M DHEA. O(2)(-) production was induced by N-formyl-methionyl-leucyl-phenylalanine and measured by the cytochrome c reduction assay. Potential scavenging of O(2)(-) by glucose, mannitol, or DHEA was assessed in a cell free system using the pyrogallol assay. MEASUREMENTS AND RESULTS: Incubation of neutrophils with glucose dose-dependently reduced O(2)(-) production, which was 50% decreased at 25 mM glucose. Also DHEA reduced the production of O(2)(-) dose-dependently, whereas production rates were unaffected by mannitol. Neither glucose, mannitol, nor DHEA scavenged O(2)(-). CONCLUSIONS: High extracellular glucose concentrations acutely reduce O(2)(-) production from activated neutrophils possibly through inhibition of G6PD. If this occurs in vivo, microbial killing by neutrophils may be impaired during acute hyperglycemia, as observed after major surgery, trauma, or severe infection. PMID- 12552366 TI - Elevated post-prandial free fatty acids are associated with cardiac sympathetic overactivity in Type II diabetic patients. PMID- 12552367 TI - Plasma concentrations of macrophage migration inhibitory factor are elevated in Pima Indians compared to Caucasians and are associated with insulin resistance. PMID- 12552368 TI - Antegrade intravenous catheterization for metabolic studies in man. PMID- 12552370 TI - [Barium examinations of the small intestine and the colon in inflammatory bowel disease]. AB - PURPOSE: This article gives an overview of the possibilities of conventional radiography in the diagnosis of inflammatory bowel disease of the small intestine and colon. MATERIAL AND METHODS: For more than 25 years we examine the small bowel employing enteroclysis with barium and methylcellulose and the colon with the usual double-contrast method. In the last 152 months 1560 small bowel enemas were performed. In the last 40 months 410 examinations of the colon were performed. RESULTS: There is a thirty percent decrease in enteroclysis examinations within the past 5 years,however, the rate of examinations with positive results increased from 46 to 57%. The proportion of the inflammatory small intestinal diseases (not only Crohn's disease) remained constant with 18%.Concerning the examinations of the colon for inflammatory disease we confirmed the diagnosis in seven cases. The radiation exposure for the enteroclysis in inflammatory diseases was 7 mSv, for colon examinations 14 mSv. CONCLUSION: Barium examinations, especially of the stomach and colon are decreasing in frequency. Therefore the art of performance and interpretation might get lost.Enteroclysis, however, is still the method of reference for the other imaging methods. The advantages compared to the other imaging methods are the excellent presentation of the details of the mucosal surface and the observation of functional disorders. PMID- 12552369 TI - [Inflammatory bowel diseases (IBD) -- critical discussion of etiology, pathogenesis, diagnostics, and therapy]. AB - AIMS: Crohn's disease and ulcerative colitis are the most frequent inflammatory bowel diseases (IBD) with a prevalence of approximately one out of 500. Cytokine research opened new and potent treatment options and thus stimulated clinical and basic research.However, the IBD still remain a challenge for patients and physicians,demanding close cooperation between gastroenterologists,radiologists and surgeons. The basic understanding of IBD,which is necessary for efficient diagnostic and therapeutic concepts is reviewed. METHODS: Based upon recent publications and our clinical experience we discuss aspects of etiology,pathogenesis,diagnostics,and therapy of Crohn's disease and ulcerative colitis. RESULTS: A genetically influenced, exaggerated and sustained immune response against the own gut flora seems to be one of the most important factors in the pathogenesis of IBD. Not less important are environmental influences. For instance, cigarette smoking had been judged to have some negative influence on the natural course of Crohn's disease.Now,however, recent studies show that smoking is even a significant independent risk factor in the pathogenesis of IBD. Since IBD and especially Crohn's disease can effect the whole body, detailed analysis of inflammatory organ involvement is necessary before therapy. For instance, the MRI enteroclysis technique adds a necessary diagnostic tool for the exploration of those parts of the small bowel that cannot been reached by routine endoscopy like the upper ileum and the lower jejunum. In terms of therapy, a change of paradigms can be observed: patients will no longer be treated only when symptoms arise, but will early be integrated into a therapeutic concept, which is determined by site and extent of the disease and adapted to the abilities and needs of the patient.Furthermore,immunosuppressive agents like azathioprine and 6 mercaptopurine will establish as central concept in the medical treatment of IBD. DISCUSSION: IBD-therapy should rather be adapted to the patient's individual inflammatory pattern than be oriented to schematic treatment rules. New endoscopic and radiologic techniques provide the necessary diagnostic tools. PMID- 12552371 TI - [Computed tomography in the diagnosis of inflammatory bowel disease -- methodology of MSCT and clinical results]. AB - PURPOSE: This paper discusses the diagnostic yield of multislice computed tomography (MSCT) in inflammatory bowel disease. METHODS: Contrast media are administered intraluminally (colon, small intestine) and intravenously (triple contrast CT). Filling of small bowel is achieved by means of jejunal tube ("Sellink CT") or via the oral route. Pharmacological relaxation of the intestine decreases motion artifact. Intraluminal contrast media consist of either hyperdense, "positive" or hypodense, "negative" liquids. RESULTS: Thin-slice MSCT of the entire abdomen allows high-quality post processing (MPR, thin-slice MIP). Due to superior distension, Sellink CT improves estimation of stenosis or changes in thickness and contrast of bowel wall. Positive contrast is superior in the detection and preoperative localization of abscess, fistula or conglomerate tumour, because it accurately differentiates between intra- and extraluminal structures.However, negative contrast facilitates quantitative evaluation of bowel wall thickening or enhancement and demonstrates gastrointestinal bleeding. CONCLUSION: MSCT of the small intestine is superior to conventional enteroclysis, especially in the diagnosis of mesenterial or other extraintestinal disease. As a side effect, the colon is assessed in the same examination. Radiation dose is less in MSCT (7.8-13.3 mSv) than in conventional fluoroscopy (13.99+/-7.57 mSv). MSCT can be performed as an alternative or adjunct to colonoscopy, if endoscopic access is restricted. It is already the imaging modality of choice in acute diverticulitis. PMID- 12552372 TI - [Follow up of Crohn's disease under therapy with hydro-MRI]. AB - PURPOSE: Evaluation of typical MRI-findings in patients with Crohn's disease receiving therapy. Correlation with the course of disease. PATIENTS AND METHODS: 81 follow-up MRI-studies in 25 patients conducted within a period of 3 weeks to 4 years were evaluated retrospectively. Therapy consisted in various combinations of antibiotics and immunosuppressive agents and if necessary operation. The findings of the MRI-studies were correlated with clinical data (e.g.operation of Crohn's complications) and the subjective perception during therapy. RESULTS: The morphological substrate of Crohn's disease in the Hydro-MRI images is reliably detected. Especially in a delineation of extraluminal changes MRI is superior to endoscopy and enteroclysis. Independent from clinical symptoms short- and mid term follow-up showed inflammatory changes of the intestinal wall in all 25 patients. In 24/81 studies there was persistence or even progression of Crohn's disease in the MRI-studies, although patients were free of symptoms by the time of image acquisition. CONCLUSION: Hydro-MRI is a modality for the evaluation of inflammatory changes in patients with Crohn's disease. Independent from clinical symptoms persistence of Crohn's disease is detectable. PMID- 12552373 TI - [MRI with oral filling in patients with chronic inflammatory bowel diseases]. AB - AIM: Evaluation of mannitol-MRI in patients with suspected or established Crohn's disease (CD). METHODS. 75 patients with suspected or established Crohn's disease were included. 1.5 l of mannitol-solution were administered orally within 1 h before imaging. A rectal filling was also employed. Butylscopolamin was applied i.v. Native-sequences were acquired. T1w sequences (axial, coronal) were acquired before and after (fs-T1-w-BH) i.v.Gd-DTPA. Additionally a dynamic CM-study was performed. RESULTS: In 45% of the examinations good image quality was achieved. In 28% opacification of the terminal ileum was insufficient. However, diagnostic assessment was possible. Motion artifacts due to breathing were rare, artifacts due to peristalsis were noted in 16% of the examinations. Alterations indicative to CD were found in 69% of the patients. The SI-increase of the thickened bowel wall was significantly higher than the increase of not thickened wall (117 vs.75%; p = 0,001 in t-test). We detected stenoses in 56%, fistulas in 23% and an abscess in one patient. CONCLUSION: Mannitol-MRI is a valuable method in the diagnostic work-up of inflammatory bowel disease. Improvement of distal distension should be attempted, because of the good acceptance of the patients and high diagnostical value. PMID- 12552374 TI - [MRI in Crohn's disease after transduodenal contrast administration using negative oral MRI contrast media]. AB - PURPOSE: To compare the efficacy and quality of conventional and MR enteroclysis with different filling methods regarding the assessment of extension and extraluminal manifestations in Crohn's disease. MATERIAL AND METHODS: 190 patients with known Crohn's disease were studied following small bowel enteroclysis, after oral administration or direct transduodenal filling in the MRI-department.T1- and T2-weighted breathhold MRI-scans w/o spectral fat suppression w/o i.v. Gd-DTPA were applied using negative oral superparamagnetic contrast media. RESULTS: Typical findings were marked bowel wall thickening with laminated wall contrast enhancement. In 135 patients 98,2% of affected bowel segments, 97,5% of stenoses and all 16 fistulas were detected, when conventional enteroclysis was employed as standard of reference. Additional important extraluminal findings such as ileoileal (n = 18), ileosigmoidal adhesions (n = 12), extraluminal abscesses (n = 35) and pseudotumors (n = 8) were visualized in 73/135 patients. Concerning the distension of jejunum and ileum, oral filling was rated significantly inferior to transduodenal filling in all small bowel segments,whereas filling in the MRI-unit was rated superior to fluoroscopic, mostly due to a mean transport time of 20 min to the MRI-unit. CONCLUSION: No clinically important findings of enteroclysis were missed when using MRI. Therefore, in patients with Crohn's disease, conventional enteroclysis can be replaced by MRI. For optimal bowel distension oral contrast administration is inferior to transduodenal filling, if a larger time delay between filling and the MRI-scan can be avoided. PMID- 12552375 TI - [The value of CT for the diagnosis of acute diverticulitis]. AB - PURPOSE: In acute diverticulitis accurate diagnosis and staging are mandatory to decide on the treatment of the patient. The impact of computed tomography (CT) on the treatment of acute diverticulitis will be discussed. MATERIAL AND METHOD: CT is performed after distension of the distal colon by means of positive,water soluble contrast media to depict intestinal perforation or penetration. Then intravenous contrast material is administered and spiral scanning is repeated to judge enhancement patterns of the abdominal structures/organs especially of the intestinal wall and to diagnose abscess formation. CT-morphologies of different stages of acute diverticulitis will be described. RESULTS: CT imaging is the only diagnostic method that in case of an acute diverticulitis combines safety with accuracy. On the one hand, it is fast and therefore safe with respect to patient control and on the other hand, it allows accurate staging of the inflammatory process reaching a sensitivity and specificity of up to 100%, each. CT is an appropriate tool to diagnose acute diverticulitis complicated by abscess formation, intestinal penetration or perforation and therefore has direct impact on the treatment of the patient. DISCUSSION: If acute diverticulitis is suspected CT is the method of choice for imaging because of its high impact on the choice of therapy and on the management of complications. PMID- 12552376 TI - [Radiological interventions in inflammatory bowel disease]. AB - INTRODUCTION: Abscesses, fistulas,hemorrhages and stenoses are common complications of inflammatory bowel diseases. This study provides an overview on various methods of radiological intervention and the clinical usefulness of these methods is analyzed. METHODS: The success rate of percutaneous abscess drainage (PAD), embolisation of hemorrhages and dilatation of bowel stenoses is reviewed and current literature is addressed. Success rate is defined in terms of cure rate and need for subsequent surgery. RESULTS: After PAD, surgery can be avoided during the observation period in about 50% of patients with abscesses due to Crohn's disease and diverticulitis. Preoperative PAD reduces the degree of invasiveness and thus the risk of surgery. Abscess recurrence is found with the same frequency following surgery or PAD. Bowel dilatation can be performed both with radiological and with endoscopic guidance.Embolisation of GI-hemorrhage is technically feasible, but the indication should be limited to strictly selected cases. CONCLUSIONS: In treating abscesses and fistulas associated with Crohn's disease and diverticulitis, PAD is a valuable treatment option. Embolisation or dilatation are restricted to rare cares. PMID- 12552377 TI - [Evaluation of radiological teaching programs in the Internet]. AB - PURPOSE: Evaluation of web based training programs, which can be contacted from the homepages of radiological departments of German universities. MATERIAL AND METHOD: From June 2000 to January 2002 the 75 web based training programs of 57 providers,which can be contacted from the web pages of the radiological departments of German universities were evaluated in a prospective study. A medical student experienced in using the world wide web examined each training program three times in an interval of six months using the following criteria: availability of the web sites, target group, kind of training program, contents and structure and the technical solution. RESULTS: 51 of the 57 the homepages were fully available at each visit. 64 of the 75 web based training programs which could be connected from these sites were available at all three visits.One program was only partially available at one spot check. 8 of the 75 programs were designed for physicians and medically trained personal, 23 were made for medical students and 44 addressed both target groups (partially more than once mentioned). The number of the presented cases ranged between one single and 3700. In 31 of 75 training programs links to other teaching files were found. A complete presentation of cases was presented by 48 of the 75 web sites.5 of the 75 web sites offered physiological images for comparison. In 20 training programs the pathological changes were optically marked in the x-ray images. A logical and didactical structure was found in 24 teaching files, 14 gave the possibility to check the learning results. No provider made use of the possibility to pass credits to the students or physicians account with regard to official training programs. Multimedia techniques were used in 15 training programs.43 sites used data reduced preview images (thumbnails). The latest update of the site is mentioned in 55 of the 75 web sites. 19 of 57 providers had either no possibility of contact or did not answer to an e-mail. CONCLUSION: From the homepages of the departments of diagnostic radiology of German universities 75 teaching files can be contacted. There is a great variety in quantity and quality. Most sites offer collections of cases. A web based training program which is comparable in quality to a CD-ROM teaching program could not be found. No program used the possibility to give credits to the users with regard to official training programs. PMID- 12552378 TI - [In memory of P.E. Peters]. PMID- 12552387 TI - [Basic principles of radiosurgical systems and their applications in arthroscopy]. AB - Electrosurgical devices are being used in each medical subdiscipline,and, only recently, they have found widespread distribution in the field of arthroscopic surgery, as well. This development is mainly due to the construction of small and relatively cheap devices that work in a bipolar mode at high frequency. A local plasma field is being built up between the electrodes at the tip of the probe which has enough energy to allow the disintegration of molecules within the treated tissue. The local current flow helps to minimize the numerous disadvantages of monopolar or Laser-based systems. Nevertheless, there is considerable discrepancy between the wide spectrum of clinical use on the one hand and the knowledge of mid- or even long-term biological effects on the treated tissue on the other. So far, clinical and experimental studies exist for the frequent indications of chondroplasty and capsular or ligamental shrinkage. But, still, there is no agreement upon these indications. Studies have shown a considerable number of dead chondrocytes or fibroblasts within a depth of several millimeters within the tissue of the joint capsule or the articular cartilage after electrosurgical treatment. But especially these cells are thought to be essential for the maintenance of the tissue. While the results of other studies seem to be less critical, some authors propose not to use electrosurgery in arthroscopy at all, until further studies prove the safety of these devises. Unfortunately, there is no reliable feedback system for the temperature increase within the treated tissue or other important parameters. The use of these instruments affords a high degree of experience from a skilled arthroscopic surgeon. PMID- 12552388 TI - [Standardisation and validation of the German version 2.0 of the Disability of Arm, Shoulder,Hand (DASH) questionnaire]. AB - The Disability of Arm, Shoulder, Hand (DASH) Version 2.0 questionnaire captures the subjective experience of patients regarding their own health. This 78-item instrument was developed by the American Academy of Orthopedic Surgeons (AAOS), the Council of Musculoskeletal Specialty Societies and the Institute for Work and Health,Toronto and measures components of health-status relevant to upper extremity conditions. We translated and validated DASH with the aim of producing a tool which could also be used in German-speaking countries. The paper reports the method of translation and cross-cultural adaptation which was carried out according to the AAOS guidelines. The result was a standardised German version of DASH. Psychometric testing of the translated questionnaire was carried out with 6 samples (n=342) who had undergone hand surgery for a variety of conditions. The study yielded good results across all groups with regard to the construct, criterion and content validity. We confirm that the German version is suitable for use in future studies. PMID- 12552389 TI - [Thoracolumbar spine fractures after conservative and surgical treatment. Dependence of correction loss on fracture level]. AB - QUESTION: This retrospective study presents results after conservative and operative treatment of thoracolumbar fractures as function of its localization. METHODOLOGY: In 2 years 70 patients with A1/A2 fracture were conservatively treated, 38 patients with A3/B/C injury were treated by internal fixtor. For evaluation 3 vertebral sections(Th5-10,Th11-L2,L3-5)were defined. Follow-up took place 1 year after implant removal or end of conservative treatment. RESULTS: The correction-loss was highest in thoracic, lowest in lumbar region. After conservative therapy,correction-loss was located to 3/4 in vertebra itself, after operative treatment especially in adjacent disc spaces. There was no general correlation to complaints. CONCLUSION: In consequence of these results A1/A2 fractures in the upper thoracic spine (15 degrees will be stabilized anteriorly, in other regions functional treated. A3-fractures of thoracic spine and thoracolumbar junction will be operated from anterior, in lower lumbar spine (>L3) from dorsal. B- and C-injuries should be instrumented with a combined dorsoventral procedure. PMID- 12552390 TI - [Possibility of sonographic early diagnosis of heterotopic ossifications after total hip-replacement]. AB - AIM: The sonographic early diagnosis of heterotopic ossifications after total hip replacement. MATERIAL AND METHOD: 53 patients were sonographically and radiologically examined after a total hip-replacement. RESULTS. In the sonographic exam,positive findings were shown in 49.1% (n=26) one week, 66% (n=35) three weeks,73.6% (n=39) six weeks and 77.3% (n=41) twelve weeks after surgery. 88.5% of patients showing positive results in the sonographic exam carried out one week after surgery, later on showed positive x-ray findings (p<0.001). Of the three-week positive x-ray findings (n=19), 78.9% had condensations in the sonographic exam (p<0.001). CONCLUSIONS: Sonographic imaging after hip-replacement seems to be a sensitive and specific method which allows an early diagnosis of ossifications one week after surgery in 2/3 of patients. In this context a therapy concept which helps avoiding the development ossifications would be welcome. PMID- 12552391 TI - [Prediction of mortality, mobility and admission to long-term care after hip fractures]. AB - INTRODUCTION: Surgical treatment, discharge planning and rehabilitation procedures are rarely based upon defined assessment procedures. It might therefore be useful to develop simple and reliable screening tools to identify patients for early discharge, intensified rehabilitation and limited treatment. PATIENTS AND METHODS: 234 patients were initially contacted. From these 217 gave informed consent. The reported study included the 134 home dwelling elderly. All patients could be contacted or died after six month. Thus,data from more than 90% could be used for the analysis. The six month mortality was 10%. A population based cohort of elderly patients referred to five local hospitals was tested. Mortality, institutionalisation and mobility were defined as major outcome criteria. Only information that was available during the first week of treatment was used in the model. Predictors expressed as odd ratios (OR) were calculated using logistic regression with variable selection. RESULTS: The most important predictors for institutionalisation were age, inability to eat without assistance, postoperative night time confusion, stroke history, cognitive deficit and fear of falling. Outdoor mobility was strongly associated to the preoperative ADL performance measured as the Barthel-Index, history of malignancy and fear of falling. Six month mortality was associated with male sex, the Barthel-Index and fear of falling. DISCUSSION: It seems feasible to improve postoperative resource allocation by predictor led stratification. This need to be tested in intervention trials under the specific condition of the German health care system. PMID- 12552392 TI - [Stabilisation of unstable trochanteric femoral fractures. Dynamic hip screw (DHS) with trochanteric stabilisation plate vs. proximal femur nail (PFN)]. AB - The dynamic hip screw (DHS) with trochanteric stabilisation plate (TSP) as the extramedullary power transmission system and the proximal femur nail (PFN) as the means of intramedullary stabilisation are both standard in the treatment of unstable trochanteric femoral fractures in the case of old people. A total of 129 patients (average age: 81,5 years) with 31 A2.2 and A2.3 as well as per /subtrochanteric femoral fractures were treated by means of osteosynthesis with DHS and TSP (n=64) or with PFN (n=65),and the results plotted in a retro /prospective study. At low complication rates, the radiological operation results are equally good. 6 revisions were necessary in the case of the DHS with TSP and 4 in the case of PFN. A significantly shorter operation time (44.3 vs. 57.3 min) and a considerably shorter in-patient stay (18.6 vs. 21.3 days) were common with PFN. The application of full-weightbearing immediately after the operation was possible for 97% of the PFN patients and 88% of the DHS patients. In a follow-up 6 months after the operation, the PFN patients displayed a significantly lower pain intensity in the operated leg at the same score for ambulation and the same subjective degree of satisfaction. Unstable pertrochanteric and per /subtrochanteric femoral comminuted fractures can be treated just as well with PFN as with DHS and TSP. Our study results,however, lead us to recommend treatment with PFN. PMID- 12552393 TI - [Results of femoral shaft fractures in childhood in relation to different treatment modalities]. AB - The clinical and radiological results of femoral shaft fractures in childhood were evaluated and compared in relation to different treatment modalities. One hundred and one children (mean age 5+/-0,4 years) were treated between 1990 to 1999. 38% of the patients were treated conservatively (mean age 2,2+/-0,5 years), 32% of the patients (mean age 6+/-0,5 years) were treated by external fixation, 17% were treated with elastic stable intramedullary nailing (ESIN, mean age 5,6+/ 0,8 years) and 12% underwent other internal fixation procedures. The duration of hospital stay was significantly longer in the conservative treatment group (18+/ 1,6 days) than in the external fixator (12+/-1,2 days) as well as in the ESIN group (8+/-0,9 days). Radiological controls revealed a significantly better reduction of the fracture by operative procedures (external fixator,ESIN) as compared to conservative management. Complications, i.e. secondary dislocation or infection, occurred in 25% of patients in the external fixator group,6% of the ESIN patients, and in 10% of the conservatively treated patients. Late complications,i.e.weight bearing dependent pain or hypertrophic scarring,were developed in both the conservative treatment group (4%) and in the external fixator group (19%); however,no late complications were seen in the ESIN group. While each method examined is known to be suitable for treatment of femoral shaft fractures in childhood,each has defined indications. Also there is a limited possibility of fracture reduction in conservative treatment this method is indicated in younger children (<4 years) where spontaneous bone remodelling is likely. For older children the ESIN method showed a low rate of complications and demonstrates the best long term results. When ESIN is not possible because of local soft tissue damage,additional injuries,or in complex fractures, the external fixator proved to be an alternative treatment for femoral shaft fractures. PMID- 12552394 TI - [Injuries to the thoracic and lumbar spine]. PMID- 12552395 TI - [Candida infection in hip alloarthroplasty]. AB - A 46-year-old polytoxicomanic patient underwent a total hip replacement for necrosis of the femoral head. The patient suffered additionally from chronic pancreatitis with insulin-dependent diabetes and polyneuropathy. Three weeks later he developed a deep wound infection followed by surgical revision. The infection persisted despite further revision operations and systemically and locally applied antibiotics. After removal of the prosthesis,microbiology revealed Staphylococcus aureus, enterococcus,and Candida parapsilosis. Five additional revision operations, application of suction-irrigation drainage, and systemically administered antibiotics could not stop the infectious process. The patient was transferred to our institution 3 months after primary surgery. The joint defect was filled with a PMMA Palacos spacer. This time biopsies were only positive for Candida albicans. After 12 days of antifungal therapy with fluconazole,microbiological biopsies were sterile. The spacer was removed and femoral extension applied. After 4 weeks of further antifungal therapy, a revision prosthesis was implanted. Until now no signs of infection have appeared. PMID- 12552396 TI - [Distinctive features of the os naviculare pseudarthrosis vs the os naviculare accessorius]. AB - We report on two cases in which the differential diagnosis between an os tibiale externum (os naviculare accessorius) and a post-traumatic pseudarthrosis of the os naviculare was subject to discussion.Both patients had suffered an acute trauma in the region of the middle foot and showed identical clinical symptoms. Conventional X-ray did not allow a definite classification of the diagnosis either.However, this was possible in both cases by virtue of typical signs evidenced by computed tomography and magnetic resonance imaging. PMID- 12552397 TI - [Heparin induced thrombocytopenia type II after the use of low-molecular weight heparin. Case report and review of the literature]. AB - INTRODUCTION: It has been suggested, that HIT-type-II can occur both after the use of low-molecular-weight and unfractionated heparin. The present study investigates the incidence of HIT-type-II after the use of low molecular weight heparin by reviewing the literature and describes one own case. CASE REPORT: We observed in a 72 year old female patient a HIT-type-II, who received low molecular weight heparin (Clexane 40 o.a.d) after the implantation of an endoprothesis as thrombosis prophylaxis. This occurred on the 9th postoperative day. One year later we performed a second endoprosthesis operation of the contralateral site without complications. With the known HIT-type-II, diagnosed the year before,we used hirudin (Refludan) for thrombosis prophylaxis. RESULT: A critical view on the international literature revealed only a few cases, where a HIT-type-II was caused by the use of low molecular weight heparin. The incidence seems to be much lower than correlated to the use of unfractionated heparin. CONCLUSION: It is possible to suffer from a HIT-type-II after low molecular weight heparin. This study confirms the suspicion that the incidence of a HIT type-II after low molecular weight is lower than after unfractionated heparin. Therefore the further use of unfractioned heparin for thrombosis prophylaxis has to be questioned and low molecular-weight heparin should be preferred. PMID- 12552398 TI - [SCIWORA-syndrome. Case report and review of the literature]. AB - The SCIWORA-syndrome was firstly described by Pang and Wilberger in 1982 [11]. It is characterized by a neurological injury without radiological appearance. Since the standardized use of MRI in spinal cord diagnosis, the number of the "real" SCIWORA-syndromes decreases. By the case report of a 14 years old boy falling down from a wall of 2 meters height with a complete paraplegia from Th12 without radiographic abnormality (even in MRI) we will give a short review of the literature. PMID- 12552399 TI - [The importance of diagnostics in faecal incontinence. Endosonography]. AB - Endoanal sonography is a well established method for the morphological diagnosis of anal sphincter damage. The best images are obtained using a 7-10 MHz rotating rigid endoprobe. The internal anal sphincter and the external anal sphincter, as well as the other pelvic floor structures, can be clearly visualised with this technique. Endosonography has shown physiological differences in sphincter anatomy and brought new insights into the pathogenesis of anorectal disorders. Apart of anal fistulas, faecal incontinence represents the main indication for the use of this method. In addition, rectal evacuation disorders are an indication for which endosonography allows a first step towards a diagnosis. Anal ultrasound is a technique friendly to both the physician and the patient, and belongs in every coloproctological unit for the assessment of faecal incontinence. Accuracy, specificity and sensitivity for the detection of anal sphincter defects range between 83 and 100% in almost all studies. Additional methods are vaginal endosonography, three dimensional endosonography and perineal sonography. PMID- 12552400 TI - [Sphincter replacement grafts]. AB - An anal sphincter replacement graft can be carried out when sphincter lesions occur after unsuccessful conservative or other treatment. Today, two different techniques are used to take over the function of the sphincter. The dynamic gracilis graft can be carried out if a non-atrophied,well innervated m. gracilis is present. This technique is carried out on patients whose incontinence is the result of a trauma,pudendopathy or imperforate anus. It can be extended to the construction of a neo-anus after abdominal resection. The artificial anal sphincter is used whenever the previous method fails or can not be used due to a non-vital, denervated or the lack of the m. gracilis. Older methods such as non stimulated gracilis, glureus or Thiersch grafts are not commonly used. PMID- 12552401 TI - [Endoanal MR imaging: diagnostic assessment]. AB - Endoanal MR imaging is an alternative to anal endosonography for the acquisition of high-resolution images of the external and internal anal sphincter. A dedicated anal receiver coil is placed in the anus so that it spans the sphincter complex. Highly detailed images of the sphincters can be obtained in any plane and the morphological abnormalities found in various types of anal incontinence can be demonstrated. Whilst MR demonstrates external sphincter disruption with an efficacy similar to that of endosonography, it is better able to demonstrate external sphincter atrophy that is presumed secondary to neuropathy. The finding of coexisting muscular atrophy on MR may prejudice the effects of anal sphincter repair for obstetric disruption. PMID- 12552402 TI - [Sacral nerve stimulation in the treatment of faecal incontinence]. AB - BACKGROUND: Sacral nerve stimulation is a recent development in the treatment of faecal incontinence. This paper reports the experiences of a single center on the use of this technique since its first application. PATIENTS AND METHOD: Based on the functional results of a temporary test stimulation phase, 16 patients underwent operative implantation of a permanent neurostimulation device. Due to a functional deficit of the morphologically intact anal sphincter, all patients suffered from incontinence for liquid and solid stool. The aetiology varied among the patients. The median duration of symptoms was 8.5 years (2-30). Clinical function was evaluated by standardized questionnaires, the Cleveland Clinic incontinence score and the disease specific quality of life instrument (FIQL ASCRS) before and during stimulation. Anorectal physiology was tested by anorectal manometry. RESULTS: Functional improvement was achieved in 94% of the patients. At a median follow-up of 32.5 months (3-99) treatment was successful in 81%. The percentage of incontinent bowel movements decreased from a median of 40% (5-100) before stimulation to 0% (0-20) with stimulation ( P=0.001). The Cleveland Clinic incontinence score improved from a median of 17 (11-20) to 5 (0 15) ( P=0.003). The quality of life index was improved in all categories.Mean squeeze pressure was increased form a median of 44 to 75 mmHg with stimulation ( P=0.003) and maximal squeeze pressure from a median of 69 to 97 mmHg (P=0.009). Resting pressure, perception, urge threshold and maximum tolerable volume were not significantly changed. CONCLUSION: Sacral nerve stimulation is an effective treatment for faecal incontinence. Morbidity is low. Sacral nerve stimulation is indicated, if conservative treatment fails and more conventional surgical approaches are of limited success. PMID- 12552403 TI - [Biofeedback in faecal incontinence]. AB - Biofeedback training is a well established method for the treatment of faecal incontinence. Prior to any biofeedback training program, a definitive diagnostic study is essential. Idiopathic faecal incontinence is the main indication for biofeedback training. Additional indications are a menacing faecal incontinence after deep anterior rectal excision with restoration of the rectal reservoir by an ileoanal pouch, anal sphincter reconstruction, rectopexy and rectocele repair. Only four studies provide evidence-based medical criteria. These, as well as numerous uncontrolled studies, show the effectiveness of biofeedback training for the treatment of faecal incontinence. Electrical stimulation of the anal sphincter is only shown to be effective in one controlled study in which it was combined with biofeedback training. PMID- 12552405 TI - [Long-term results after Spitzy's umbilical hernia repair]. AB - A total of 140 patients presenting with umbilical hernia underwent Spitzy's operation and were included in a retrospective study to analyse the recurrence rate. The patients received a questionnaire and were invited for a physical examination including ultrasound. Participation included 108 patients (follow-up 77%). A total of seven patients (6.5%) developed postoperative wound infections. Prolonged postoperative impairment was mentioned by nine patients (8.3%). After a mean period of 16 days, the patients were able to continue their occupation, and after 27 days, they could manage their usual physical activity. The recurrence rate was 13.0%. The risk for a recurrence correlated with the relative bodyweight. In the group with a body-mass-index (BMI) of less than 30, the recurrence rate was 8.1 %, whereas 31.8% of the patients with a BMI of more than 30 developed a recurrent hernia. The size of the hernial orifice also had an influence on the postoperative result. In patients with a hernial gap smaller than 1 cm the recurrence rate was 6.3%, with hernia orifices from 1 to 2 cm the rate was 4.1%, from 2 to 3 cm 14.3%, from 3 to 4 cm 25.0% and in patients with hernial gaps greater than 4 cm the rate of recurrences was 54.5%. Considering these results,we recommend the use of alloplastic material for umbilical hernia repair for patients with a BMI greater than 30.0 and hernia orifice larger than 3 cm. The decision for use of a mesh in hernial gaps from 2 to 3 cm should depend on individual factors. PMID- 12552404 TI - [Neoadjuvant therapy in non-small cell lung cancer. Prognostic impact of "mediastinal downstaging"]. AB - In the course of a prospective multicenter study, 40 (26 squamous cell and 14 adenocarcinomas) patients with stage IIIA and IIIB non-small cell lung cancer (NSCLC) were submitted to surgery after neoadjuvant radiochemotherapy. Pretherapeutic clinical lymph node status was compared to the lymph node involvement established in the resection specimens. Therapy-induced tumor regression was classified according to a three-step tumor regression grading system. In 29 patients (72.5%) a downward shift in lymph node involvement could be established,whereas in 27.5% ( n=11) pretherapeutic lymph node status was maintained. Of 26 patients with post-therapeutic N0 or N1 status, 21 revealed less than 10% vital tumor tissue in the resection specimens (regression grades IIb or III). Patients with post-therapeutic N0 or N1 lymph node status were found to have a survival benefit compared to patients with N2 lymph node involvement, though this difference was not statistically significant (p=0.27). On the other hand, tumor regression showed a significant correlation to the overall survival period (p=0.02). Thus, therapy-induced tumor regression grading seems to be a more precise method to predict the outcome of the disease. PMID- 12552406 TI - [Prognosis of malignant melanoma following dissection regional lymph node metastases]. AB - BACKGROUND: Malignant melanomas usually metastasize in the regional lymph nodes. This generally leads to a worsening of prognosis. Survival probability after extirpation of the tumorous regions is conditioned by various factors. Until now, the basis for differentiated therapy has been the precise understanding of the TNM classification. Clinical parameters are also relevant to therapy and of decisive importance to the further course of disease. QUESTION: Which clinical parameters are important to the prognosis of patients with regional melanoma and lymph node disease? PATIENTS AND METHODS: This study is a retrospective analysis of patients at the Clinic and Polyclinic of General Surgery of the University Hospital in Munster,Germany, together with a review of the literature. Included were 137 patients who received curative resection of lymph node metastases from malignant melanomas between 1974 and 1996. From the literature, 6,694 cases were found which allowed the establishment of 13 different prognosis parameters following lymph node metastasis resection These parameters were compared with our own results concerning established, relevant parameters. RESULTS: The cumulative 5-year survival rate for patients studied was 50.7%. The 5-year survival rates relating to characteristics varied between 12.9% and 80.0%. Of the 13 variables from the literature relevant to prognosis that were included in this analysis, four were found to be significant: Breslow penetration depth, standardized age and age at primary tumor diagnosis, recurrence, and location of the primary tumor. Considering these significant characteristics, one can arrive at a detailed prognostic classification of patient and history. This is indispensable for correctly tailoring therapy to disease stage. PMID- 12552407 TI - [Receptors and signal proteins of the innate immune system as new targets in sepsis therapy. Reprogramming of the immune response]. AB - During polymicrobial sepsis,microbial pathogens and their products activate the innate immune system through signaling receptors of the Toll-like receptor (TLR) family, resulting in hyperinflammation and organ injury. The analysis of preclinical mouse models has shown that inactivation of the common TLR signaling adaptor protein MyD88 prevents the hyperinflammatory response and improves survival.Importantly, MyD88 deficiency does not impair antibacterial defense mechanisms.Thus,TLRs and proteins involved in TLR signaling may represent interesting targets for the development of new drugs for reprogramming pathophysiological immune responses during sepsis. PMID- 12552408 TI - [Glomus tumour of the stomach. A case report]. AB - Glomus tumours are benign neoplasms that usually arise in the skin of the extremities but have infrequently been found to occur in other sites including the stomach. We report on a 71-year-old female with non-specific epigastric pain who was diagnosed as having a small, intramural gastric tumour in addition to a cholecystolithiasis. Intraoperatively, the tumour was investigated by frozen section, but the diagnosis remained inconclusive. The ultimate histological examination showed clusters of uniform epithelioid cells surrounding wide vascular spaces. This led to the diagnosis of a glomus tumour. In a review of the recent literature,we discuss the methods and limitations of preoperative diagnostic measures. PMID- 12552409 TI - [Squamous cell carcinoma of the liver. A rare diagnosis in the course of intrahepatic cholangiolithiasis]. AB - We report the rare case of a squamous cell carcinoma of the liver in a 69-year old female Asian suffering from cholangitis and a liver abscess due to segmental intrahepatic cholangiolithiasis. The patient was an immigrant who had been living for several years in Austria. After pigtail drainage of an abscess in segment II and III, a bisegmentectomy was performed. Histopathological examination of the specimen showed structures of an invasive squamous cell carcinoma close to the resection margin.For that reason we performed a left lobectomy. Seven years thereafter the patient is well and free of local and distant recurrences. The case is discussed based on a review of the literature. PMID- 12552410 TI - [The new medical curriculum: a chance for the psychosocial disciplines]. PMID- 12552411 TI - [Teaching of psychosomatic medicine and psychotherapy as an element of the Dresden DIPOL-Curriculum -- the PBL-course "Nervous system" and psyche]. AB - A federal law, the "Approbationsordnung fur Arzte", regulates Medical education in Germany. In 2002 a revised version of this law demanded of the medical faculties to implement interdisciplinary and problem-based (PBL) courses in their curriculum. Already in 1998 the faculty of medicine of the TU Dresden decided to reform the traditional curriculum. The reform-curriculum is being created in close co-operation with the Harvard Medical School. It is a hybrid curriculum retaining traditional elements as lectures and bedside teaching with PBL tutorials and a special doctor-patient-communication-training, which is based on role-play exercises. Psychosomatic Medicine is taught together with neurology and psychiatry as a major topic of the 7,5-week PBL-course "Nervous System and Psyche" but there are also lectures and special skills training exercises in other PBL-courses like "Oncology" or "Emergency medicine". Data of the external evaluation showed, that the students accepted psychosomatic topics of the curriculum very well. This paper describes concept, implementation and evaluation data of the PBL-course "Nervous System and Psyche". PMID- 12552412 TI - [Integration of a doctor/patient-communication-training into clinical teaching. Examples from the reform-curricula of Heidelberg and Dreden Universities]. AB - It is an important demand of educational reforms to improve the skills of the communication with patients. To reach this goal neither interdisciplinary courses nor tutorials with paper-cases are sufficient. This is the reason why teaching modules including a training to improve doctor-patient-communication were integrated into the reform-curricula at the universities of Heidelberg (Heidelberger Curriculum Medicale, HEICUMED) and Dresden (Dresdner Integrated Patient-Oriented Learning, DIPOL). These modules were originated in cooperation of psychosomatic/psychotherapeutic clinics with representatives of other departments, which provides a chance to integrate psychosomatic issues into the teaching of other disciplines and to promote basic psychosomatic competences. The concept, the realization, and the results of the evaluation of these teaching modules will be described. The evaluation showed that the students rated them as mainly positive, as more effective than traditional learning and that the training improved their self-efficacy related to communicative competences. PMID- 12552413 TI - [Integrative psychosomatics: contributions to a reform of medical training]. AB - Medical training has so far mainly emphasised teaching of knowledge and neglected fostering of theoretical and practical competence in doctors' occupation. The consequences of these deficits are often experienced as "practice shock" when young doctors start working. New concepts of teaching moduls in psychosocial subjects may enhance medical competence. We present some of the contents and didactic concepts which we have implemented in the medical curriculum of Cologne University. PMID- 12552414 TI - [Comprehensive evaluation of medical teaching -- a task for the psychosocial disciplines?]. AB - The paper describes the experiences with a comprehensive evaluation of medical teaching at the medical faculty of the Friedrich-Schiller-University Jena. Based upon evaluations of courses initiated by the students, the evaluation of teaching is organized scientifically and extended as a comprehensive evaluation of several relevant issues (e. g. surveys of teachers and alumni) by the staff of the Department of Medical Psychology since 1998. Originally supported by a grant of the state of Thuringia, the evaluation is now financed by the medical faculty. The paper describes the requested resources for staff and material to establish a systematic evaluation of medical teaching. In addition, a detailed concept of the allocation of financial means considering evaluations, examinations, and other indicators of teaching quality is demonstrated. Based upon the results of studies related to the evaluation data base, the question of the establishment of an external evaluation in view of the coming changes of the medical curriculum in Germany is critically discussed. PMID- 12552415 TI - [Improving medical education in psychosocial disciplines by multimedia tools]. AB - The use of multimedia tools is gaining increasing importance in medical education. These tools provide adequate techniques and possibilities for education within all medical including the psychosocial disciplines. Until now, a lack of multimedia learning programs and websites providing material related to psychosocial issues has to be considered in Germany. On the other hand, a new Medical Licensure Act (Approbationsordnung fuer Aerzte) will have a dramatic impact on medical education and teaching. On this basis there is a broad need for multimedialearning-environments, covering parts of the changes demanded by the new educational standards. Considering the lack of available multimedia learning programs and the need for those tools due to the medical education reform in Germany, the rapid development, validation and curricular implementation of those programs is required. This should be a goal of high priority, especially for the teaching of issues related to medical psychology, medical sociology, and psychosomatic medicine and psychotherapy. PMID- 12552416 TI - [Chronic diseases, psychological distress and coping -- challenges for psychosocial care in medicine]. AB - Due to the increase of chronic diseases within the last decades the need and demand for psychosocial treatment in medicine has been realized. This review focuses on the psychosocial aspects of chronic diseases and discusses selected topics of medical and rehabilitation psychology. Recent developments in quantitative and qualitative methods have allowed the systematic analysis of psychosocial distress and coping with chronic disease as well as the consequences on social relationships. The need for psychosocial treatment in acute care and rehabilitation can be diagnosed by differential assessment tools for coping and psychiatric morbidity. Specific approaches of psychology and psychotherapy for patients with somatic diseases have been developed and may be regarded as an integrative part of medical treatment in acute care and rehabilitation. In rehabilitation, the traditional individualistic view of psychotherapy has been broadened towards vocational integration and participation in social activities as outcome criteria. Evaluation research as well as the rehabilitation sciences have provided empirical data on psychosocial treatment of chronically ill patients. Under increasing financial restrictions and problems of the health care systems there is a need for quality assurance and the proof of scientific evidence to guarantee psychosocial treatment as an integrated part of medical care in the future. PMID- 12552417 TI - Increased risk of mother-to-infant transmission of hepatitis C virus by intrapartum infantile exposure to maternal blood. AB - Virological and clinical data from 73 hepatitis C virus (HCV)-infected pregnant women who gave birth to 75 children were merged retrospectively, by logistic regression analysis, to investigate risk factors for vertical transmission of HCV. Eighty-two percent of the HCV-infected mothers were HCV-RNA-positive during pregnancy, and 10% were coinfected with human immunodeficiency virus (HIV). Nine children were HCV infected, 1 was HIV infected, but none was HIV-HCV coinfected. Among vaginal deliveries, the mean HCV load of mothers who transmitted HCV to their infants was higher than that of those who did not (8.1 x 10(5) vs. 1.4 x 10(4) copies/mL; P=.056). A reduction in umbilical cord-blood pH (relative risk, 3.9; P=.04) or the occurrence of perineal or vaginal laceration (relative risk, 6.4; P=.028) during vaginal delivery significantly increased the risk of vertical HCV transmission. In conclusion, high maternal viremia, infantile hypoxia, and intrapartum exposure to virus-contaminated maternal blood increased the risk of HCV transmission during vaginal deliveries. Consequently, cesarean section may reduce the risk of vertical HCV transmission in selected cases. PMID- 12552418 TI - Molecular epidemiology and forensic genetics: application to a hepatitis C virus transmission event at a hemodialysis unit. AB - Molecular phylogenetic analyses are frequently used in epidemiologic testing, although only occasionally in forensics. Their acceptability is hampered by a lack of statistical confidence in the conclusions. However, maximum likelihood testing provides a sound statistical framework for the testing of phylogenetic hypotheses relevant for forensic analysis. We present the results of applying this method to a small hepatitis C outbreak produced in a hospital hemodialysis unit that involved 6 patients. Polymerase chain reaction products from a 472-nt fragment of the E1-E2 region, including the hypervariable region, HVR-1, of the hepatitis C virus genome were cloned, and an average of 10 clones/patient and from 11 additional control patients were sequenced. The method allows a statistical evaluation that the likelihood of each sample belonging or not to a given group, a question of relevance in many forensic and epidemiological analyses of molecular sequences. PMID- 12552419 TI - Human herpesvirus 8: seroprevalence and correlates in prostitutes in Mombasa, Kenya. AB - Human herpesvirus 8 (HHV-8) infection is very prevalent in sub-Saharan Africa, but the role of sexual transmission has not been well characterized. HHV-8 seroprevalence and correlates were evaluated in a cohort of female prostitutes in Mombasa, Kenya. Between February 1993 and January 2000, stored plasma samples taken from 736 women were tested, by whole-virus ELISA assay, for the presence of HHV-8 antibodies; of these 736 women, 633 were included in the analysis of correlates of HHV-8 infection; and, of these 633, 44.1% were seropositive for HHV 8 antibodies. In univariate analysis, age, years of education, years of prostitution, workplace, hormonal contraception, intrauterine-device use, alcohol consumption, syphilis, and gonorrhea were all significantly associated with the presence of HHV-8 antibodies. In a multivariate model, older age, fewer years of education, and 2 markers of high-risk sexual behavior-namely, alcohol consumption and gonorrhea-were each independently associated with HHV-8 seropositivity. These results suggest that heterosexual transmission may contribute to acquisition of HHV-8 infections in this African population of prostitutes. PMID- 12552420 TI - Antigen burden is major determinant of human immunodeficiency virus-specific CD8+ T cell maturation state: potential implications for therapeutic immunization. AB - The majority of untreated human immunodeficiency virus (HIV) type 1-infected individuals ultimately develop uncontrolled viremia and progressive disease. Cytotoxic T lymphocytes (CTLs) are known to play an important role in controlling HIV-1 replication, which has led to an increasing interest in augmenting conventional antiretroviral therapy with therapeutic vaccination. The successful development of a therapeutic vaccine will rely on the ability to correlate an aspect of the immune response with clinical outcome. In this study, the CD8(+) T cell maturation status of antigen-specific cells in models of well and poorly controlled virus infections were compared, to show that a memory phenotype predominates when antigen loads are absent or low. In HIV-1 infection, the emergence of memory CD8(+) T cells was found to occur only in individuals with highly suppressed viral replication for an extended duration. Such assessments of the immune response may provide a refined measure of virus control. PMID- 12552421 TI - Cell-associated genital tract virus and vertical transmission of human immunodeficiency virus type 1 in antiretroviral-experienced women. AB - To determine the association between genital tract shedding of human immunodeficiency virus (HIV) type 1 and vertical transmission, a case-control substudy was conducted within the Women and Infants Transmission Study. Antenatal cervicovaginal lavage specimens were assessed for HIV-1 RNA in the supernatant and HIV-1 RNA and DNA in cell pellets. Multivariate analyses compared 26 women who transmitted HIV to their infants with 52 women who did not; 33% received combination antiretroviral therapy, and 65% received monotherapy. Adjusted odds ratios (ORs) for the presence (OR, 3.42; 95% confidence interval [CI], 0.76-15.4; P=.11) and titer (OR, 1.66; 95% CI, 0.93-2.99; P=.09) of HIV-1 DNA suggested that there is an independent association with vertical transmission. When analyses were restricted to vaginal and nonelective cesarean deliveries, each one-log increase in mean titer of HIV-1 DNA was associated with a significantly higher risk of transmission (OR, 2.28; 95% CI, 1.09-4.78; P=.03). The cell-associated genital tract compartment is important in the pathophysiology and prevention of vertical HIV-1 transmission. PMID- 12552422 TI - Combination antiretroviral therapy results in a rapid increase in T cell receptor variable region beta repertoire diversity within CD45RA CD8 T cells in human immunodeficiency virus-infected children. AB - Human immunodeficiency virus (HIV) type 1 disrupts the T cell receptor (TCR) variable region (V) beta repertoire in CD8 T cells by impairing thymic capacity and skewing postthymic cellular maturation. The TCR repertoire was examined using spectratyping of CDR3 length diversity within CD45RA and CD45RO CD8 T cells in HIV-infected and healthy children. In healthy children, CDR3 lengths displayed Gaussian distribution in both CD45RA and CD45RO subsets. Vbeta families in HIV infected children displayed a large proportion of perturbations in both subsets. High virus load and advanced immunosuppression correlated with increased perturbations within CD45RA but not CD45RO CD8 T cells. After therapy and virus suppression, there was rapid reestablishment of Gaussian distributions in CD45RA cells. HIV-1-induced disruption of TCR diversity within CD45RA CD8 T cells correlates with disease progression. Suppression of viral replication by treatment results in the rapid correction of TCR diversity in this CD8 subset because of emergence of new T cells from the thymus. PMID- 12552423 TI - Selective modulation of superantigen-induced responses by streptococcal cysteine protease. AB - Streptococcal pyrogenic exotoxin (Spe) B, a streptococcal cysteine protease, is believed to be important in group A streptococcal (GAS) pathogenesis. The present study examined the effect of SpeB on the activity of superantigenic exotoxins secreted by M1T1 GAS isolates. The proliferative response of human lymphocytes to culture supernatant (SUP) from an SpeB(+) isolate increased significantly (P<.05) when the isolate was grown with N-[N-(L-3-trans-carboxyoxirane-2-carbonyl)-L leucyl]-agmatine, a cysteine protease inhibitor. The lymphocyte-stimulating activity of SUP from a spontaneous SpeB(-) variant or SpeB(-) knockout (DeltaSpeB) mutant was also significantly higher than that of SUP from the SpeB(+) parent isolate (P<.001). The addition of recombinant SpeB to the DeltaSpeB mutant reduced the lymphocyte response to a level comparable to that with the SpeB(+) isolate. SpeB affected superantigens that stimulate cells expressing T cell receptor Vbeta (TCRBV)-4, TCRBV7, and TCRBV8 but not those that stimulate TCRBV2. SpeB has a selective proteolytic effect on GAS superantigens. PMID- 12552424 TI - Activation of platelet gpIIbIIIa by phospholipase C from Clostridium perfringens involves store-operated calcium entry. AB - Clostridium perfringens gas gangrene is characterized by rapid tissue destruction, and amputation remains the single best treatment. Previous studies have demonstrated that tissue destruction follows C. perfringens phospholipase C (PLC)-induced, platelet gpIIbIIIa-mediated formation of occlusive intravascular platelet/leukocyte aggregates. In this study, the intracellular signaling events leading to activation of gpIIbIIIa by PLC were investigated. PLC activated surface expressed gpIIbIIIa and mobilized gpIIbIIIa from internal stores. Chelation of intracellular calcium or inhibition of store-operated calcium entry each blocked PLC-induced activation of gpIIbIIIa, whereas inhibition of protein kinase C was without effect. Thus, PLC initiates an "inside-out" signaling cascade that begins with depletion of internal calcium stores, is sustained by an influx of calcium through store-sensitive channels, and culminates in the functional activation of gpIIbIIIa. These findings suggest that calcium-channel blockade and strategies targeting gpIIbIIIa may prevent vascular occlusion, maintain tissue viability, and provide an alternative to radical amputation for patients with gas gangrene. PMID- 12552425 TI - Inheritance of susceptibility to induced Escherichia coli bladder and kidney infections in female C3H/HeJ mice. AB - In the present study, the inheritance of resistance and susceptibility to bladder and kidney infections in BALB/c, C3H/HeJ, F(1), and backcross mice was investigated, and the number of genes contributing to the phenotypes was estimated. Infections were induced in female mice by intravesical inoculation with Escherichia coli, and the number of bacteria in bladder and kidneys was quantified at 10 days. The (BALB/c x C3H/HeJ) F(1) mice had bladder and kidney infection intensities equivalent to those observed in the resistant BALB/c parents. Twelve percent of the (F(1) x C3H/HeJ) backcross mice had severe bladder infections, similar to the susceptible C3H/HeJ parents. Kidney infections ranging in intensity between those observed in BALB/c and C3H/HeJ parents were present in one-half of the backcross mice. Statistical analyses indicated that >/=1 gene is responsible for the increased susceptibility of C3H/HeJ mice and that the trait appears to be recessive. PMID- 12552426 TI - Chlamydia trachomatis infection alters host cell transcription in diverse cellular pathways. AB - To study the responses of the host cell to chlamydial infection, differentially transcribed genes of the host cells were examined. Complementary DNA (cDNA) probes were made from messenger RNAs of HeLa cells infected with Chlamydia trachomatis and were hybridized to a high-density human DNA microarray of 15,000 genes and expressed sequence tags. C. trachomatis alters host cell transcription at both the early and middle phases of its developmental cycle. At 2 h after infection, 13 host genes showed mean expression ratios >/=2-fold. At 16 h after infection, 130 genes were differentially transcribed. These genes encoded factors inhibiting apoptosis and factors regulating cell differentiation, components of the cytoskeleton, transcription factors, and proinflammatory cytokines. This indicates that chlamydial infection, despite its intravacuolar location, alters the transcription of a broad range of host genes in diverse cellular pathways and provides a framework for future studies. PMID- 12552427 TI - Cell-to-cell contact of human monocytes with infected arterial smooth-muscle cells enhances growth of Chlamydia pneumoniae. AB - Chlamydia pneumoniae can infect arterial cells. It has been shown that coculture of human monocytes (U937) and endothelial cells promotes infection of C. pneumoniae in endothelial cells and that the enhancement was mediated by a soluble factor (insulin-like growth factor 2) secreted by monocytes. In this study, it is shown that coculture of monocytes with C. pneumoniae enhances infection of C. pneumoniae in arterial smooth-muscle cells 5.3-fold at a monocyte to-smooth-muscle cell ratio of 5. However, unlike endothelial cells, no enhancement was observed if monocytes were placed in cell culture inserts or if conditioned medium from monocyte cultures was used, which suggests that cell-to cell contact is critical. The addition of mannose 6-phosphate or octyl glucoside, a nonionic detergent containing a sugar group, to cocultures inhibited the enhancement. These findings suggest that the monocyte-smooth-muscle cell interaction may be mediated by mannose 6-phosphate receptors present on monocytes. PMID- 12552428 TI - Mortality due to gastroenteritis of unknown etiology in the United States. AB - Gastroenteritis of unknown etiology (GUE) is a significant cause of mortality in the United States. In the present study, the demographic and medical characteristics of people who died of GUE were examined, using the 1995-1997 Multiple Cause of Death files to calculate GUE death rates and proportionate mortality ratios. There were 13,153 GUE deaths during the period, or approximately 4400 deaths per year. Death rates were highest among infants and elderly persons, especially nursing home residents, and increased during the winter months. Compared with all decedents, GUE decedents were more likely to have certain other medical conditions, including bacteremia, volume depletion, renal failure, and human immunodeficiency virus/acquired immunodeficiency syndrome. Fatal GUE often appeared to be infectious in origin, but death certificates provide insufficient information to determine whether the causative agents were unknown or foodborne. The accuracy of GUE reporting on death certificates and the etiology of fatal GUE merit further investigation. PMID- 12552429 TI - A murine model of coccidioidal meningitis. AB - Coccidioidal meningitis is lethal in humans. A reproducible murine model was established by lumbar intrathecal injection of Coccidioides immitis arthroconidia. Cerebrospinal fluid (CSF) samples were obtained by cisternal puncture. Lethal infection developed in all mice given 10-60 colony-forming units (cfu). Lethargy, ataxia, or paralysis preceded death. Temporal studies after challenge with 27 cfu revealed positive brain (4/5 mice) and spinal cord (2/5 mice) cultures on day 3; CSF samples contained 688 leukocytes/mm(3) and 33 cfu/mL. The results of histopathologic analysis were unremarkable. By day 8, all mice were culture positive (5.0 log(10) cfu in brain tissue and 4.1 log(10) cfu in spinal cord tissue); CSF samples contained 4833 leukocytes/mm(3) and 3425 cfu/mL. Histopathologic examinations showed acute meningitis of the brain and spinal cord, some parenchymal invasion and abscesses, and meningeal arteritis. Groups of mice given ketoconazole had prolonged survival and suppressed lung disease; histopathologic examination demonstrated granulomatous meningitis, possibly a more chronic form. With the development of these models, studies of pathogenesis, host response, and therapy are possible. PMID- 12552430 TI - Platelet accumulation in brain microvessels in fatal pediatric cerebral malaria. AB - The pathogenesis of fatal cerebral malaria (CM) is not well understood, in part because data from patients in whom a clinical diagnosis was established prior to death are rare. In a murine CM model, platelets accumulate in brain microvasculature, and antiplatelet therapy can improve outcome. We determined whether platelets are also found in cerebral vessels in human CM, and we performed immunohistopathology for platelet-specific glycoprotein, GPIIb-IIIa, on tissue from multiple brain sites in Malawian children whose fatal illness was severe malarial anemia, CM, or nonmalarial encephalopathy. Platelets were observed in 3 locations within microvessels: between malaria pigment and leukocytes, associated with malaria pigment, or alone. The mean surface area of platelet staining and the proportion of vessels showing platelet accumulation were significantly higher in patients with CM than in those without it. Platelet accumulation occurs in the microvasculature of patients with CM and may play a role in the pathogenesis of the disease. PMID- 12552431 TI - Treatment history and treatment dose are important determinants of sulfadoxine pyrimethamine efficacy in children with uncomplicated malaria in Western Kenya. AB - This study retrospectively studied amendable determinants of sulfadoxine pyrimethamine (SP) efficacy involving 2869 treatments among 1072 Kenyan children <5 years old who had uncomplicated malaria. The dose was based on age: one quarter tablet was given to infants <1 year old, one-half tablet was given to 1-3 year-old children, and a full tablet was given to 4-year-old children. Only 23.5% received the internationally recommended target dose of 25/1.25 mg of SP per kg of body weight. SP intake in the previous 15-35 days (adjusted relative risk, 1.67; 95% confidence interval, 1.35-2.07) and low SP dose (<27.5/1.375 mg/kg) (adjusted relative risk, 1.58; 95% confidence interval, 1.17-2.13) explained 38% of parasitological treatment failures by day 7. Patients with recent SP intake are likely to have recrudescent infections and may need close follow-up if treated with SP or alternative treatment. Applying our weight-for-age data to 31 existing age-based SP dose recommendations predicted that 22 of them would result in underdosing of >25% of children <5 years. Many age-based dose recommendations need urgent revision, because SP is increasingly used as first-line treatment in sub-Saharan Africa. PMID- 12552432 TI - Common surface-antigen var genes of limited diversity expressed by Plasmodium falciparum placental isolates separated by time and space. AB - Plasmodium falciparum placental parasites from Cameroon have been shown to express surface variant var genes encoding Duffy binding-like (DBL)-gamma domains that bind chondroitin sulfate A. All 5 domains exhibited sequences with 39%-55% amino acid (aa) identities and appear sufficiently conserved to function in receptor binding. Transcripts of 2 samples showed complete conservation over 4 kb, demonstrating for the first time distinct conserved placental var genes. Four placental isolates from Gabon collected 4 years later expressed DBL-gamma sequences with 85%-99% aa identities to those from Cameroon, confirming the conserved nature of placental variants separated by time and location. Five peripheral parasites from children also displayed DBL-gamma sequences with 75% 97% homologies. From this, it can be concluded that P. falciparum parasites expressing unique var DBL-gamma genes can cause placental malaria, referred to as varPAM genes. This demonstration of structurally/functionally constrained DBL gamma chondroitin sulfate A-binding domains is relevant to understanding pregnancy-associated malaria pathogenesis and to vaccine development. PMID- 12552433 TI - Hookworm aspartic protease, Na-APR-2, cleaves human hemoglobin and serum proteins in a host-specific fashion. AB - Hookworms are voracious blood-feeders. The cloning and functional expression of an aspartic protease, Na-APR-2, from the human hookworm Necator americanus are described here. Na-APR-2 is more similar to a family of nematode-specific, aspartic proteases than it is to cathepsin D or pepsin, and the term "nemepsins" for members of this family of nematode-specific hydrolases is proposed. Na-apr-2 mRNA was detected in blood-feeding, developmental stages only of N. americanus, and the protease was expressed in the intestinal lumen, amphids, and excretory glands. Recombinant Na-APR-2 cleaved human hemoglobin (Hb) and serum proteins almost twice as efficiently as the orthologous substrates from the nonpermissive dog host. Moreover, only 25% of the Na-APR-2 cleavage sites within human Hb were shared with those generated by the related N. americanus cathepsin D, Na-APR-1. Antiserum against Na-APR-2 inhibited migration of 50% of third-stage N. americanus larvae through skin, which suggests that aspartic proteases might be effective vaccines against human hookworm disease. PMID- 12552434 TI - Does antigen-specific cytokine response correlate with the experience of oculorespiratory syndrome after influenza vaccine? AB - During the 2000-2001 season in Canada, a newly identified oculorespiratory syndrome (ORS) was observed in patients after immunization with inactivated influenza vaccine. ORS was associated with a high proportion of microaggregates of unsplit virions in the implicated vaccine and had clinical features suggesting delayed-onset hypersensitivity. We explore the association between in vitro cytokine balance (type 1 vs. type 2) and clinical ORS after influenza vaccination. We report the balance of interferon (IFN)-gamma, interleukin (IL) 10, IL-5, and IL-13 expression by peripheral blood mononuclear cells (PBMC) among unvaccinated, vaccinated ORS-affected, and vaccinated ORS-unaffected persons after in vitro challenge with implicated and nonimplicated vaccines. Antigen stimulated PBMC from vaccinated persons produced significantly more IFN-gamma than did those from unvaccinated persons. There was a statistically significant type 2 polarization among unvaccinated compared with vaccinated persons. Although vaccinated ORS-affected individuals had less of a type 1 basis than did vaccinated unaffected individuals, this difference was not statistically significant. PMID- 12552435 TI - A passive immunotherapy, (PE)HRG214, in patients infected with human immunodeficiency virus: a phase I study. AB - (PE)HRG214 (HRG) is a polyclonal antibody preparation produced by immunization of goats with purified human immunodeficiency virus (HIV) antigens. In this phase I study, HRG was administered intravenously as a single dose (1, 2, 4, 8, or 16 mg/kg) to 18 HIV-1-infected patients with CD4 cell counts >/=50 cells/microL and virus loads >/=500 copies/mL. The most frequent adverse event was a transient rash, which appeared to be both dose- and CD4 cell count-dependent. At the 16 mg/kg level, median half-life was 68.4 h, and median C(max) was 392 microg/mL, a level well above that which inhibits HIV in vitro. At that dose level, median and maximum decreases in HIV-1 RNA levels at day 8 were 0.24 log(10) and 0.58 log(10), respectively, and, at day 29, were 0.24 log(10 ) and 2.2 log(10), respectively. HRG, administered as a single dose, is reasonably well tolerated and achieves adequate plasma concentrations. PMID- 12552436 TI - Effect of GB virus C coinfection on response to antiretroviral treatment in human immunodeficiency virus-infected patients. AB - To study how GB virus C (GBV-C) coinfection affects the response to highly active antiretroviral therapy (HAART), 146 human immunodeficiency virus (HIV)-infected patients were tested for GBV-C RNA and antibodies against GBV-C E2 protein, and responses to HAART were evaluated. GBV-C-infected patients exhibited a complete virological response to HAART more often than patients without [correction] GBV-C and had a greater increase in median CD4 cell count and a marginally greater median HIV RNA level decrease. This association was found to be independent of baseline CD4 cell count and plasma HIV RNA, which indicates that an association exists between GBV-C infection and response to HAART. PMID- 12552437 TI - A potential virulence gene, hylEfm, predominates in Enterococcus faecium of clinical origin. AB - An open reading frame (hyl(Efm)) with homologies to previously described hyaluronidase genes has been identified in nonstool isolates of Enterococcus faecium. E. faecium isolates (n=577) from diverse sources were screened for the presence of hyl(Efm) and esp(Efm), a putative virulence gene associated with epidemic E. faecium strains. The presence of esp(Efm) was roughly twice that of hyl(Efm), but both were found primarily in vancomycin-resistant E. faecium isolates in nonstool cultures obtained from patients hospitalized in the United States. These data suggest that specific E. faecium strains may be enriched in determinants that make them more likely to cause clinical infections. Differences in the prevalence of these strains may help explain variations in the clinical importance of multiresistant E. faecium across different continents. PMID- 12552438 TI - Lipoarabinomannan-reactive human secretory immunoglobulin A responses induced by mucosal bacille Calmette-Guerin vaccination. AB - The ability of 17 recombinant mycobacterial proteins, native antigen 85 complex, lipoarabinomannan (LAM), and Mycobacterium tuberculosis lysate to detect antibody responses induced by bacille Calmette-Guerin (BCG) vaccination and active tuberculosis infection were studied in enzyme-linked immunosorbent assays. Only LAM-reactive serum immunoglobulin G responses were significantly increased in both BCG-vaccinated patients and patients with active tuberculosis (P<.05), and oral BCG vaccination also induced significant increases in LAM-reactive secretory immunoglobulin A (P<.05). LAM-reactive antibody assays can serve as markers of humoral and mucosal immunity in future trials of BCG and newer attenuated mycobacterial vaccines. PMID- 12552439 TI - Detection of Ureaplasma urealyticum in second-trimester amniotic fluid by polymerase chain reaction correlates with subsequent preterm labor and delivery. AB - Ureaplasma urealyticum is the microorganism most frequently isolated from the amniotic fluid of women in preterm labor. The relationship between intra-amniotic U. urealyticum in healthy second-trimester pregnant women and subsequent pregnancy outcome was investigated. Transabdominal amniotic fluid obtained from 254 asymptomatic women at 15-17 weeks' gestation were tested by polymerase chain reaction (PCR). U. urealyticum was identified in 29 subjects (11.4%). A subsequent preterm labor occurred in 17 U. urealyticum-positive women (58.6%), compared with 10 (4.4%) U. urealyticum-negative women (P<.0001). Preterm birth was documented in 7 (24.1%) U. urealyticum-positive women compared with only 1 U. urealyticum-negative woman (0.4%) (P<.0001). U. urealyticum-positive women also had a higher prevalence of preterm labor in a prior pregnancy (20.7%) than did the negative women (2.7%; P=.0008). PCR testing of second-trimester amniotic fluid for U. urealyticum can identify women at risk for subsequent preterm labor and delivery. PMID- 12552440 TI - Loss of red blood cell-complement regulatory proteins and increased levels of circulating immune complexes are associated with severe malarial anemia. AB - Severe anemia is one of the most lethal complications of Plasmodium falciparum malaria. Red blood cells (RBCs) from children with severe malarial anemia are deficient in complement regulatory proteins (CR1 and CD55). A case-control, age- and sex-matched study was carried out to determine whether these deficiencies are acquired or inherited and the relative contribution of these complement regulatory protein deficiencies, the immune complex level, and the parasite density to the development of severe malarial anemia. RBC CR1 and CD55 deficiencies resolved after treatment, suggesting that these changes were acquired. Using conditional logistic regression, a decline in CD55 (or CR1) (odds ratio [OR], 4.2; 95% confidence interval [CI], 2.1-8.1; P<.001) and an increase in immune complex level (OR, 2.6; 95% CI, 1.5-4.8; P=.001) were significantly associated with severe malarial anemia. PMID- 12552441 TI - Herpes simplex virus infection of dendritic cells: balance among activation, inhibition, and immunity. AB - Several lines of evidence suggest that dendritic cells (DCs), the most potent antigen-presenting cells known, play a role in the immunological control of herpes simplex virus (HSV) infections. HSV infection of DCs induced submaximal maturation, but DCs failed to mature further in response to lipopolysaccharide (LPS). LPS induced interleukin (IL)-12 secretion, and the induction of primary and secondary T cell responses were impaired by infection. Ultimately, DC infection resulted in delayed, asynchronous apoptotic cell death. However, infected DCs induced HSV recall responses in some individuals. Furthermore, soluble factors secreted by DCs after infection induced DC maturation and primed for IL-12 secretion after LPS stimulation. These data support a pathogenetic model of HSV infection, in which initial delay in the generation of immune responses to HSV at peripheral sites is mediated by disruption of DC function but is overcome by bystander DC maturation and cross-presentation of HSV antigens. PMID- 12552442 TI - Human herpesvirus (HHV)-6 and HHV-7: two closely related viruses with different infection profiles in stem cell transplantation recipients. AB - Human herpesvirus (HHV)-6 and HHV-7 loads were evaluated retrospectively in peripheral blood mononuclear cells (PBMC) from 78 recipients of stem cell transplantation (SCT) by real-time polymerase chain reaction. The median HHV-6 load in patients was 1357 genome equivalent copies (EqCop)/10(6) PBMC but was below the quantitation threshold in 31 immunocompetent individuals, which strongly suggests that HHV-6 reactivation occurred after SCT. The HHV-6 load was higher in patients with delayed neutrophil engraftment (P=.002) or severe graft versus-host disease (P=.009). Moreover, the occurrence of at least 1 HHV-6 related manifestation (fever, cutaneous rash, pneumonitis, or partial myelosuppression) was statistically associated with a concomitant virus load >10(3) EqCop/10(6) PBMC (P=.007). Conversely, HHV-7 reactivation was not favored, because median HHV-7 loads were similar in patients and healthy control subjects (1053 vs. 1216 EqCop/10(6) PBMC). The kinetics of Roseolovirus loads during the posttransplantation period suggested that HHV-7 may act as a cofactor of HHV-6 reactivation. PMID- 12552443 TI - Homologous versus heterologous immune responses to Norwalk-like viruses among crew members after acute gastroenteritis outbreaks on 2 US Navy vessels. AB - Host immune responses to human caliciviruses are difficult to study because of the lack of a clear definition of antigenic or serological types. This report describes antibody responses to several Norwalk-like viruses in large outbreaks of acute gastroenteritis on 2 US Navy ships. Enzyme immunoassays (EIAs) were used to measure antibody responses. To understand the antibody response to a homologous strain causing the outbreaks, the viral capsid gene of one isolate (C59) was expressed in baculovirus and included in the EIAs. Significantly greater seroresponses were detected in patients against the homologous strain than against the heterologous strains. Strains within genogroups reacted more strongly than did strains between genogroups. Significantly higher antibody titers against the outbreak strain were detected in acute serum samples from control subjects than in those from case patients. These results indicate that recombinant EIAs are useful for outbreak investigation and that the homologous antibody might be protective against reinfection. PMID- 12552444 TI - Serum immunoglobulin G response to human papillomavirus type 16 virus-like particles in human immunodeficiency virus (HIV)-positive and risk-matched HIV negative women. AB - Baseline serum samples from 2815 human immunodeficiency virus (HIV)-positive and 963 HIV-negative women enrolled in 2 cohort studies were tested for immunoglobulin G antibodies to human papillomavirus type 16 (HPV-16) capsids. HPV 16 seropositivity was associated with lifetime number of sex partners (P<.001) among both HIV-positive and HIV-negative women. Approximately 50%-60% of HPV-16 DNA-positive women were HPV-16 positive. HPV-16 seropositivity was associated with HIV infection; however, after adjustment for baseline cervical HPV infection and disease, the association disappeared. Thus, the high seroprevalence of HPV-16 among HIV-positive women may be explained by a high prevalence of HPV of all types. Approximately 50% of HIV-positive women had serological evidence of prior HPV-16 infection, but only approximately 5% had an HPV-16 cervical infection at baseline. Despite the higher prevalence of HPV infection in this group, most HIV positive women are able to control HPV-16 replication at the cervix, and reactivation, if it occurs, is not very common. PMID- 12552445 TI - Cellular human immunodeficiency virus (HIV)-protective factors: a comparison of HIV-exposed seronegative female sex workers and female blood donors in Abidjan, Cote d'Ivoire. AB - Cellular factors that may protect against human immunodeficiency virus (HIV) infection were investigated in 27 HIV-exposed seronegative (ESN) female sex workers (FSWs) and 27 HIV-seronegative female blood donors. Compared with blood donors, ESN FSWs had significantly decreased expression levels of C-X-C chemokine receptor 4 (CXCR4), but not of C-C chemokine receptor 5, on both memory (P<.001) and naive (P=.041) CD4(+) T cells. CXCR4 down-regulation was associated with prolonged duration of commercial sex work by ESN FSWs. CD38 expression on CD8(+) T cells was significantly increased among ESN FSWs, compared with that among blood donors (P=.017). There were no differences in HLA-DR and CD62L expression between blood donors and ESN FSWs. Proportions of T cells producing the beta chemokines RANTES (regulated on activation, normally T cell-expressed and secreted), macrophage inflammatory protein (MIP)-1alpha, and MIP-1beta or the cytokines interleukin (IL)-2, IL-4, interferon-gamma, and tumor necrosis factor alpha, were similar in the 2 groups. These data indicate that ESN FSWs differ from HIV-seronegative female blood donors with respect to immunological factors that have no clear protective potential against HIV transmission. PMID- 12552446 TI - Limited protective effect of the CCR5Delta32/CCR5Delta32 genotype on human immunodeficiency virus infection incidence in a cohort of patients with hemophilia and selection for genotypic X4 virus. AB - The relationship among CCR5 genotype, cytomegalovirus infection, and disease progression and death was studied among 159 human immunodeficiency virus (HIV) infected patients with hemophilia. One patient (0.6%) had the CCR5Delta32/CCR5Delta32 genotype (which occurs in approximately 2% of the Scandinavian population) and a rapid disease course. His HIV V3 region contained genotypic features attributable to X4 virus and resembled functionally verified X4 virus and virus from patients treated with a CD4 cell-stimulating drug, tucaresol. Age-related differences in disease progression rate and survival time were seen for CCR5/CCR5 patients. Surprisingly, no protective effect of the CCR5/CCR5Delta32 genotype on disease progression or survival was seen for children but was evident for adults. Age group-related immunologic differences might explain this variation, and transmission route and/or viral phenotype variation within donor virus may be related to the limited protection of the CCR5Delta32/CCR5Delta32 genotype. Sequence comparisons indicate that X4 virus can be selected in vivo due to either absence of CCR5 receptors or relative increase of CXCR4 receptors. PMID- 12552447 TI - Moving to human immunodeficiency virus type 1 vaccine efficacy trials: defining T cell responses as potential correlates of immunity. AB - There is evidence in both simian immunodeficiency virus and human immunodeficiency virus (HIV) type 1 infection that class I major histocompatibility complex-restricted CD8(+) cytotoxic T lymphocytes play a pivotal role in controlling infection and, potentially, in protecting by immunization. Progress has been made in designing strategies to elicit these responses with HIV-1 vaccines, but methods to reproducibly quantify them have posed difficulties. An interferon-gamma enzyme-linked immunospot assay, using peptide pools spanning the HIV-1 genes, was developed and standardized. This method is rapid (2 days), sensitive (threshold of detection, > or =0.005%), quantitative, feasible using cryopreserved cells, and able to define epitope specificities. When this assay was applied to 36 HIV-1-seropositive and 10 HIV-1 seronegative subjects, it proved to be robust (specificity, 100%). When responses in natural infection were compared with vaccine-induced responses, vaccine recipient responses were > or =1 log lower, which confirms the importance of using this sensitive assay as an initial screen in vaccine protocols. PMID- 12552448 TI - Purified protein derivative-activated type 1 cytokine-producing CD4+ T lymphocytes in the lung: a characteristic feature of active pulmonary and nonpulmonary tuberculosis. AB - Because tuberculosis (TB) is primarily a pulmonary disease, we examined the cytokine responses of CD4(+) T lymphocytes in bronchoalveolar lavage (BAL) fluid after incubation with purified protein derivative (PPD) in human immunodeficiency virus-negative patients with TB and control subjects with nontuberculous respiratory disease. Parallel blood and BAL fluid samples from each subject were incubated with or without PPD, and the proportions of CD4(+) T lymphocytes producing interferon (IFN)-gamma or tumor necrosis factor (TNF)-alpha were measured by flow cytometry. The proportions of PPD-activated IFN-gamma- and TNF alpha-producing CD4(+) cells were low among control subjects (median, 0.33% and 0.78%, respectively). By contrast, among patients with TB, strong IFN-gamma and TNF-alpha responses were demonstrated (median, 24.0% and 32.4%, respectively), regardless of whether the TB was pulmonary or nonpulmonary. Measurement of type 1 cytokine production by CD4(+) T lymphocytes in response to PPD in BAL fluid is a promising new diagnostic test for active TB in immunocompetent individuals. PMID- 12552449 TI - Multidrug resistance among Enterobacteriaceae is strongly associated with the presence of integrons and is independent of species or isolate origin. AB - This study investigated the extent to which multidrug resistance (MDR) among Enterobacteriaceae is related to DNA elements called "integrons," whether the relationship is species dependent or origin dependent, and which resistance patterns are associated with integrons. Analysis of 867 nonrepeat isolates comprising 8 species and originating from the community and 23 European hospitals showed a significant relation between MDR and integrons, independent of species or origin. Although resistance to each tested antimicrobial agent was significantly associated with integrons, only resistance to sulfamethoxazole, cotrimoxazole, gentamicin, tobramycin, ampicillin, piperacillin, and cefuroxime predicted the presence of integrons. Combined resistance to both ampicillin and sulfamethoxazole-trimethoprim was the starting point for the development of resistance to additional beta-lactams, aminoglycosides, cephalosporins, and ciprofloxacin, a development paralleled by an increasing prevalence of integrons. The acquisition of resistance genes is not random, and the transfer of integron carrying elements plays a dominant role in the development of MDR by Enterobacteriaceae. PMID- 12552450 TI - Campylobacter transmission in a Peruvian shantytown: a longitudinal study using strain typing of campylobacter isolates from chickens and humans in household clusters. AB - Campylobacter jejuni is a major cause of pediatric diarrhea in developing countries-free-ranging chickens are presumed to be a common source. Campylobacter strains from monthly surveillance and diarrhea cases were compared by means of restriction-fragment length polymorphism (RFLP), rapid amplified polymorphic DNA, and Lior serotyping. RFLP analysis of 156 human and 682 avian strains demonstrated identical strains in chickens and humans in 29 (70.7%) of 41 families, and 35%-39% of human isolates from diarrhea and nondiarrhea cases were identical to a household chicken isolate. Isolation of the same RFLP type from a household chicken and a human within 1 month was highly protective against diarrhea (odds ratio, 0.07; P<.005). Campylobacter strains from symptomatic humans were unlikely to be identical to strains recently carried by household chickens, limiting the potential benefits from household-based control measures. PMID- 12552451 TI - Whole blood bactericidal activity during treatment of pulmonary tuberculosis. AB - The timely evaluation of new drugs that can be used to shorten tuberculosis (TB) treatment will require surrogate markers for relapse. This study examined bactericidal activity against intracellular Mycobacterium tuberculosis in whole blood culture (whole blood bactericidal activity; WBA) during TB treatment. In the absence of chemotherapy, immune mechanisms in patient blood resulted in bacteriostasis, whereas administration of oral chemotherapy resulted in bacillary killing. Total WBA per dose was greater during the intensive phase of treatment than during the continuation phase (mean, -2.32 vs. -1.67 log(10) cfu-days, respectively; P<.001). Cumulative WBA throughout treatment was greater in subjects whose sputum cultures converted to negative by the eighth week of treatment than in those for whom conversion was delayed (mean, -365 vs. -250 log(10) cfu-days; P=.04) and correlated with the rate of decrease of sputum colony-forming unit counts during the first 4 weeks of treatment (P=.018), both of which are indicative of prognosis. These findings indicate that measurement of WBA may have a role in assessing the sterilizing activity of new anti-TB drugs. PMID- 12552452 TI - Chlamydia trachomatis omp1 genotypic diversity and concordance with sexual network data. AB - Sexual and social network analysis have been proposed as novel sexually transmitted disease control and research tools. Here, the concordance between chlamydia genotype data and a large sexual network constructed from routinely collected contact tracing data was examined. A sexual network was constructed for Manitoba, Canada, from province-wide contact tracing data. Positive chlamydia specimens from the same time period were collected and genotyped by omp1 DNA sequencing. A high degree of concordance was found between transmission events, on the basis of molecular data, and proposed transmission events, on the basis of sexual network data. Discordant results appeared to occur when a portion of the network contained potential core group members or in areas where contact tracing is difficult to carry out. The agreement between the molecular and epidemiologic data suggests that the use of routine contact tracing data is a valid approach for the construction of sexual networks. PMID- 12552453 TI - Lipopolysaccharide-binding protein serum levels in patients with severe sepsis due to gram-positive and fungal infections. AB - Lipopolysaccharide-binding protein (LBP) is increased in patients with severe gram-negative infections, but LBP serum levels have not been reported for in patients with gram-positive and fungal infections. LBP serum levels were determined in patients with severe sepsis secondary to gram-positive or fungal infections and were compared with LBP serum levels obtained from patients with gram-negative mixed infections and from healthy volunteers. Thirty-seven episodes of severe sepsis were analyzed among 24 patients. LBP serum levels were significantly increased in patients with severe sepsis (46.4+/-28.3 microg/mL), compared with that of healthy volunteers (5.7+/-1.9 microg/mL; P<.0001). On the other hand, LBP serum levels obtained from patients with gram-negative infections (40.80+/-34.79 microg/mL) did not differ from those obtained from patients with gram-positive (35.55+/-23.95 microg/mL) or fungal (39.90+/-22.19 microg/mL) infections. These data suggest that LBP is an aspecific marker of sepsis, and the response was not clearly correlated with severity. Furthermore, in patients with multiple episodes of sepsis, LBP response seems to be of lesser magnitude after each subsequent episode of severe sepsis. PMID- 12552454 TI - Modeling of Plasmodium falciparum-infected erythrocyte cytoadhesion in microvascular conditions: chondroitin-4-sulfate binding, A competitive phenotype. AB - Although chondroitin-4-sulfate (CSA) is expressed throughout the microvasculature and CSA-binding infected erythrocytes (IE(CSA)) cytoadhere to lung and brain endothelial cells and sequester in male Saimiri sciureus, this phenotype seems to be dependent on the presence of a placenta to develop. This contradiction was investigated by modeling the interactions and cytoadhesion parameters in the microvasculature. Mixtures of IEs interacting with CSA, CD36, or intercellular adhesion molecule 1 were incubated with endothelial cells expressing the corresponding receptors, at physiological pH, under flow conditions. By use of suspensions composed of equal proportions of the phenotypes, cytoadhesion of approximately 10 times as many IE(CSA) as of any other IE tested was observed. Adherent IE(CSA) resisted microvascular wall shear stresses 3-15 times more effectively than did the others. These results, which require confirmation with field isolates, demonstrate that the CSA phenotype is competitive and are consistent with this phenotype initiating microvessel occlusion and with CSA mediated sequestration in microvessel conditions. PMID- 12552455 TI - A waterborne outbreak of Norwalk-like virus among snowmobilers-Wyoming, 2001. AB - In February 2001, episodes of acute gastroenteritis were reported to the Wyoming Department of Health from persons who had recently vacationed at a snowmobile lodge in Wyoming. A retrospective cohort study found a significant association between water consumption and illness, and testing identified Norwalk-like virus (NLV) in 8 of 13 stool samples and 1 well. Nucleotide sequences from the positive well-water specimen and 6 of the positive stool samples were identical. This multistrain NLV outbreak investigation illustrates the importance of NLV as a cause of waterborne illness and should encourage monitoring for NLVs in drinking water. PMID- 12552456 TI - SEN virus infection in patients with chronic hepatitis C: preferential coinfection with hepatitis C genotype 2a and no effect on response to therapy with interferon plus ribavirin. AB - To clarify the influence that a recently identified SEN virus (SENV) has on hepatitis C virus (HCV) response to therapy with interferon plus ribavirin, 2 SENV variants, SENV-D and SENV-H, were studied in 100 patients with chronic hepatitis C; 57 of these patients were positive for SENV-D/H DNA, and there were no differences, in clinicopathological features, between patients with and without SENV coinfection. However, patients with SENV coinfection had a higher prevalence of HCV genotype 2a than did those without it. The sustained HCV response rate after combination therapy was comparable between patients with and without SENV coinfection. Of the 57 patients with SENV coinfection, 18 (32%) had a sustained SENV response to combination therapy, and SENV-D had a higher sustained response rate than did SENV-H. These results suggest that SENV has a specific link to HCV genotype 2a and that SENV infection has no apparent effect on coexisting chronic hepatitis C. PMID- 12552457 TI - Adenovirus DNA in serum of children hospitalized due to an acute respiratory adenovirus infection. AB - Serum samples from 68 immunocompetent infants (mean age, 12.6 months) with an acute adenovirus infection of the respiratory tract (39 experiencing their first adenovirus infection) were tested for the presence of adenovirus DNA, to investigate whether viral dissemination via the blood is usually present in the immunocompetent patient. Using a nested polymerase chain reaction assay, adenovirus DNA could be detected in acute-phase serum samples from 28 (41%) children. Adenovirus DNA was never found in follow-up serum samples, indicating a short period ( approximately 1 week) of viral dissemination. In children experiencing their first adenovirus infection, viral DNA could be detected in 72% of the acute-phase serum samples collected within the first week after onset of symptoms. Adenovirus DNA could also be detected in 25% of the acute-phase serum samples from patients with reinfection. PMID- 12552458 TI - Priming of human immunodeficiency virus type 1 (HIV-1)-specific CD8+ T cell responses by dendritic cells loaded with HIV-1 proteins. AB - Proteins may serve as ideal CD8(+) T cell immunogens for human immunodeficiency virus type 1 (HIV-1) if they can be delivered to and processed through the human leukocyte antigen class I pathway. This study shows that human blood monocyte derived dendritic cells loaded with liposome-complexed HIV-1 proteins and matured with CD40 ligand can prime CD8(+) T cells to HIV-1 in vitro. Whole HIV-1 protein in liposome may be an effective immunogen for HIV-1 vaccine protocols. PMID- 12552459 TI - Interleukin-2 Increases CD4+ lymphocyte numbers but does not enhance responses to immunization: results of A5046s. AB - To ascertain whether CD4(+) lymphocyte increases induced by interleukin (IL)-2 enhanced in vivo immune responses, 38 human immunodeficiency virus (HIV)-infected patients who had received highly active antiretroviral therapy (HAART) or HAART and IL-2 for at least 60 weeks were immunized with tetanus toxoid, inactivated glycoprotein 120-depleted HIV-1, and hepatitis A and B vaccines. Despite dramatic increases in CD4(+) lymphocyte counts, IL-2 did not enhance immunization responses. PMID- 12552460 TI - Polymorphism of SLC11A1 (formerly NRAMP1) gene confers susceptibility to Kawasaki disease. AB - Since its first description in Japan >30 years ago, Kawasaki disease (KD) has been reported worldwide. Although an infectious etiology is suspected based on the epidemiology and clinical features, a causative agent has not been identified. The disease is more frequent in children of Japanese ancestry, and siblings of children with KD have a significantly greater risk of developing KD than do children of the same age in the general population. This suggests a possible genetic susceptibility to KD. Results of this study showed that allele 1 of the 5' promoter (GT)n repeat in the SLC11A1 (formerly NRAMP1) gene, which endows the gene with a weak promoter activity, was highly represented in patients with KD. This suggests possible explanations for both the infectious etiology of this disease and the genetic risk in the Japanese population. PMID- 12552461 TI - Immune response to capsular polysaccharide and surface proteins of Streptococcus pneumoniae in patients with invasive pneumococcal disease. AB - The immune response to pneumococcal capsular polysaccharides (CPSs) and to the pneumococcal surface proteins cell wall-associated serine proteinase A (PrtA), pneumococcal surface protein A (PspA), and Streptococcus pneumoniae pullulanase A was evaluated in 45 patients with invasive pneumococcal disease compared with healthy adults. In serum from patients with meningitis and pneumonia, CPS antibody levels were low, compared with healthy adults; antibody levels did not differ between groups and did not change between phases. Levels of immunoglobulin G directed against the investigated pneumococcal surface proteins in patients with invasive pneumococcal disease were in the same range as in healthy adults. However, median PrtA and PspA antibody levels tended to increase during early convalescent phase. Low levels of CPS antibody, rather than of antibodies directed against the pneumococcal surface proteins, may predispose to invasive pneumococcal infection. PMID- 12552462 TI - The CagA protein of Helicobacter pylori is translocated into epithelial cells and binds to SHP-2 in human gastric mucosa. AB - Recent experiments have indicated that CagA of Helicobacter pylori is injected into epithelial cells via the type IV secretion system and undergoes tyrosine phosphorylation in cells and that translocated CagA binds the SRC homology 2 domain-containing tyrosine phosphatase (SHP-2). We investigated these phenomena in in vivo human gastric mucosa. Tyrosine-phosphorylated CagA and CagA coimmunoprecipitated SHP-2 were detected in gastric mucosa from H. pylori positive patients with atrophic gastritis and in noncancerous tissues from H. pylori-positive patients with early gastric cancer. In contrast, CagA was not detected in gastric mucosa with either intestinal metaplasia or cancer. Our results provide the first evidence that CagA is translocated into the gastric epithelial cells, receives tyrosine phosphorylation, and binds SHP-2 in in vivo human gastric mucosa. Deregulation of SHP-2 by CagA may play a role in the acquisition of a cellular-transformed phenotype at a relatively early stage of multistep gastric carcinogenesis. PMID- 12552463 TI - Transforming growth factor-beta, interleukin (IL)-18, and IL-12: effect on the clinical course and complications of Plasmodium falciparum malaria. PMID- 12552465 TI - Clinical value of adjunctive interleukin-2: clarifications and corrections. PMID- 12552467 TI - The role that the functional Asp299Gly polymorphism in the toll-like receptor-4 gene plays in susceptibility to Chlamydia trachomatis-associated tubal infertility. PMID- 12552469 TI - The abdominal compartment syndrome following aortic surgery. AB - BACKGROUND: multi-organ failure is a leading cause of death following aneurysm surgery, especially in the emergency setting. Intra-abdominal hypertension is an important factor in the development of multi-organ failure. Prevention, early recognition and prompt treatment of abdominal hypertension and the abdominal compartment syndrome may reduce mortality following aneurysm surgery. METHODS: a descriptive review of the literature from a Medline search. RESULTS AND CONCLUSIONS: the abdominal compartment syndrome is the result of diverse physiological effects caused by increased intra-abdominal pressure. The syndrome has been most widely described in trauma victims, but occurs in patients following aortic surgery, particularly following ruptured aneurysm repair. Preventative therapy should be instituted to minimise its development in patients at risk, and monitoring of intra-abdominal pressure may allow prompt treatment of this condition. PMID- 12552470 TI - Do vascular surgery patients need a cardiology work-up? A review of pre-operative cardiac clearance guidelines in vascular surgery. AB - OBJECTIVES: to outline the appropriate pre-operative cardiac work-up for patients who are scheduled for major peripheral vascular surgery. DESIGN: review of the literature. MATERIALS AND METHODS: a review of the literature focusing on studies that have correlated the pre-operative cardiac work-up patients receive to the cardiac morbidity and mortality following vascular surgery. Only studies with level A evidence were included. RESULTS: peri-operative beta blockade has been shown to decrease cardiac complications after vascular surgery in all risk groups. Non-invasive cardiac testing is only necessary for patients in the intermediate/high risk group. Coronary revascularization should only be considered after a positive non-invasive cardiac test. CONCLUSIONS: patients must be risk stratified pre-operatively based on history and physical examination. Low risk patients should receive peri-operative beta blockade only with no further non-invasive testing. On the other hand, intermediate and high risk patients should undergo non-invasive cardiac testing before going to the operating room. PMID- 12552471 TI - Risk factors of early and late complications in patients undergoing endovascular aneurysm repair. AB - OBJECTIVES: to identify pre-operative factors that could predict complications following from transluminal repair of abdominal aortic aneurysms (AAA). METHODS: during a 5-year period, 96 consecutive patients underwent elective endovascular treatment of a AAA. In all patients, helical CT and/or Magnetic Resonance Imaging (MRI), and plain abdominal roentgenogram were performed at 1, 3, 6, 12, 18, and 24 months and yearly thereafter. Angiography was performed systematically 1 year after the stent-graft implantation, or earlier if helical CT or MRI diagnosed an increase in the maximal transverse diameter or a high flow endoleak. RESULTS: early (<30 days) morbidity (12%) was significantly increased by pre-operative renal insufficiency (p < 0.01). Early mortality (2%) correlated with ASA score (p = 0.01). Median follow-up was 27 months (range 3-66). Mortality (12%) during follow-up was correlated to the pre-operative coronary status (p = 0.01). A type I endoleak was diagnosed in 18 patients (19%). Common iliac artery diameter was correlated with the presence of type I endoleak (p < 0.001). A type II endoleak was diagnosed in 47 (49%) patients. The diagnostic of type II endoleak was significantly increased (p = 0.001) in patients with pre-operative patent IMA associated with more than four patent lumbar arteries. The anatomic characteristics of the aneurysm were correlated to the additional endovascular procedures during stentgraft implantation (p = 0.01), and to the implantation of a complementary iliac limb extension during follow-up (p = 0.01). CONCLUSIONS: the risk factors determined by this statistical analysis could help surgeons to select more accurately patients suitable for endovascular treatment. PMID- 12552472 TI - Techniques for the endovascular management of complications following lower limb percutaneous transluminal angioplasty. AB - OBJECTIVES: to determine the incidence of early complications following percutaneous transluminal angioplasty and to describe their management and outcome. MATERIALS: five hundred and fifty consecutive patients undergoing angioplasty of 648 limbs, containing 1053 anatomical segments during a two year period were reviewed retrospectively. RESULTS: early complications affected 109 segments (10%) in 92 limbs (14%) of 84 patients (15%). Of the 109 segments affected by early complications, 106 (97%) were managed by endovascular techniques with surgery being required on only three (3%) occasions. There were no deaths attributable to angioplasty. CONCLUSIONS: although early complications occur in 14% of limbs undergoing percutaneous transfemoral angioplasty, the majority (97%) can be managed by endovascular techniques. PMID- 12552473 TI - A 1 year prospective study of management and outcome of patients presenting with critical lower limb ischaemia. AB - OBJECTIVES: to determine management of patients with critical lower limb ischaemia (CLI) from first presentation to investigation and treatment. DESIGN: prospective study of critical ischaemia patients. METHODS: one-year prospective survey (May 2000-May 2001). Follow-up 3-15 months. RESULTS: some 873 arterial cases presented, 134 patients had CLI. Of the latter 49% were men, 30% diabetic, the median age was 81 years. Only 15 (24%) of 62 cases were referred to outpatients as urgent. Patients waited a median of 25 days (range 1-100) to be seen in outpatients, and had symptoms for a median of 8 weeks. Treatment was conservative for 70 patients, and 11 primary amputations, six secondary amputations, and 62 revascularisation procedures (34 operative, 28 percutaneous transluminal angioplasty) were performed. At follow-up (3-15 months, median 9 months), rates of major amputation and death were 12 and 27% respectively. Significantly more diabetics underwent major amputation (p < 0.02) than non diabetics. Patients presenting with ulceration or gangrene were at greater risk of death than those with rest pain alone (p < 0.01). CONCLUSION: patients with CLI often have symptoms for many weeks before being seen by a specialist, and 76% are referred as non-urgent cases. This compares with patients with suspected malignant disease in the U.K. who are required to be seen within 2 weeks. PMID- 12552474 TI - Vascular training in the U.K.: femorodistal bypass, an index procedure? AB - OBJECTIVES: femorodistal bypass operation is one of three index procedures for vascular training in the U.K. Our aim is to determine the suitability of femorodistal bypass to be considered as an index procedure in the era of increasing utilisation of percutaneous transluminal angioplasty (PTA). DESIGN: a retrospective analysis of prospectively collected data. PATIENTS AND METHODS: a total of 526 patients with 608 chronic critically ischaemic limbs admitted to the vascular unit, at the Royal United Hospital, Bath, between January 1994 and December 1999 was included in the study. RESULTS: revascularisation either by PTA, bypass surgery or a combination of both was attempted in 524 limbs (86%). Crural procedures were carried out on 71 limbs (14% of revascularised limbs). Primary crural procedures included 34 PTAs as a sole treatment (48%), and 37 femorodistal bypass operations (52%). CONCLUSIONS: during a 6-year period only 37 primary femorodistal bypass operations were performed in a unit which aggressively treats CLI. PTA is the initial step and increasingly the sole treatment for critical limb ischaemia (CLI), including distal lesions. We question the rationale of including an uncommon operation as a vascular training index procedure. PMID- 12552475 TI - Comparison of Magnetic Resonance Angiography (MRA) and Duplex Ultrasound Arterial Mapping (DUAM) prior to infrainguinal arterial reconstruction. AB - OBJECTIVE: the purpose of this study is to compare Magnetic Resonance Angiography (MRA) to Duplex Ultrasound Arterial Mapping (DUAM) and intraoperative findings to determine the clinical, accuracy of MRA for planning lower extremity revascularization procedures. METHODS: forty-two patients who underwent lower extremity revascularization procedures had both MRA and DUAM evaluations. These data were analyzed retrospectively and two distinct endpoints were considered. First, we compared the MRA and DUAM findings for aorto-iliac, femoral-popliteal and infra-popliteal segments to intraoperative findings and evaluated the degree to which they agreed. Second, if there was a disagreement between imaging modalities, it was established whether a change in operative procedure would have resulted. RESULTS: MRA and DUAM findings agreed in 26 of 31 cases (83%) of aorto iliac segments, in 25 of 31 cases (81%) of femoral-popliteal segments, and in 16 of 21 cases (76%) of infra-popliteal segments. In total, DUAM agreed with intraoperative findings in 98% of cases while MRA agreed in 82% (p<.001). Disagreement between intraoperative findings and DUAM lead to an alternate surgical procedure in only one case (2%) while disagreement with MRA lead to a different procedure in 38% of cases (p<.001). CONCLUSIONS: these data show that MRA is not yet adequate to replace conventional angiography and is less accurate that DUAM. Further improvements are necessary before MRA can be used as the sole modality for formulation of a pre-operative plan for lower extremity revascularization. PMID- 12552476 TI - The acquisition of methicillin-resistant Staphylococcus aureus (MRSA) in vascular patients. AB - OBJECTIVE: the aim of this study was to establish at which point during a hospital admission MRSA acquisition occurs in vascular patients. METHOD: a consecutive series of 100 patients undergoing arterial surgery were screened for MRSA carriage on admission to hospital, on exit from theatre, on discharge from ITU, weekly whilst an inpatient and on hospital discharge. Screening was with moistened swabs from nose, throat, perineum and open wounds that were pooled for microbiological culture. RESULTS: four patients (4%) screened positive for MRSA on admission to hospital. Of the remaining 96, 16 (17%) acquired MRSA during their hospital stay as follows: exit from theatre, one; exit from ITU, six; on the ward postoperatively, nine. Comparing MRSA acquisition (n=16) with non acquisition (n = 80) the following characteristics were noted, length of stay 16 (4-66) vs 7 (2-50) days (Mann-Whitney p < 0.001); admission to ITU 13/16 vs 46/80 (Fishers chi-squared p = 0.10); length of ITU stay 3 (1-20) vs 3 (1-14) days (Mann-Whitney p = 0.41). Frequent hospital attendance, age, emergency admission, diabetes or renal failure were not commoner in those with MRSA acquisition. CONCLUSIONS: these data indicate that 4% of patients undergoing arterial surgery are pre-existing carriers of MRSA. Length of hospital stay is the single most important determinant of MRSA acquisition. PMID- 12552477 TI - Peripheral and central arterial haemodynamic interactions are early abnormalities in young male cigarette smokers. AB - BACKGROUND: smoking, result in endothelial dysfunction and this affects systemic and local haemodynamics. The aim was to assess interactions of the left ventricle and arterial system of smokers at baseline and after a physiological stimulus- the cold pressor test (CPT) which causes a sympathetically driven vasoconstriction that counteracts the normal endothelial dependent vasodilatation. MATERIALS AND METHODS: male smokers and controls were compared using applanation tonometry. Parameters included systolic and diastolic blood pressure, ejection duration, heart rate, aortic augmentation index (AAI), and sub endocardial viability ratio (SEVR). The CPT was performed at 1 and 3 min following immersion of the hand in ice. RESULTS: smokers have abnormal baseline cardiac timing (heart rate and ejection duration), systolic and diastolic blood pressures which are due to increased peripheral wave reflection (AAI) and thus affect the SEVR. Following CPT, the pressure wave differential, dP/dt, was significantly increased in smokers compared to non-smokers who had a decrease at 1 min in ice. Mean systolic and diastolic pressure was significantly increased in both groups at 1 and 3 min as was end systolic pressure in non-smokers. CONCLUSIONS: baseline ventriculo-vascular dynamics, are abnormal as was the evoked response to CPT. The blunted blood pressure increase of smokers compared to controls following CPT, may represent altered nitric oxide production in the macro and microcirculation through differential upregulation of endothelial nitric oxide synthase (eNOS) and inducible NOS respectively. The potential for therapeutic intervention and prevention of ongoing endothelial injury, requires further investigation. PMID- 12552478 TI - Durability of reflux-elimination by a minimal invasive CHIVA procedure on patients with varicose veins. A 3-year prospective case study. AB - OBJECTIVES: to assess the outcome of a conservative and haemodynamic method for insufficient veins on an ambulatory basis (French acronym, "CHIVA") with preservation of the greater saphenous vein (GSV) for treatment of primary varicose veins. METHODS: duplex incompetence of the sapheno-femoral junction (SFJ) and the GSV trunk, with the re-entry perforating point located on a GSV tributary was demonstrated in 58 patients with varices (58 limbs). The re-entry point was defined as the perforator, whose compression of the superficial vein above its opening eliminates reflux in the GSV. Duplex scanning was performed preoperatively and at 7 days, and patients were followed prospectively at 1, 3, 6, 12, 24, and 36 months after CHIVA. Operation consisted in flush ligation and division from the GSV of the tributary containing the re-entry perforating vein (no additional high ligation is included). If reflux returned, SFJ interruption was performed in a second surgical procedure. RESULTS: the GSV diameter showed an average reduction from 6.6 to 3.9 mm 36 months after surgery. Reflux in the GSV system was demonstrated in all but five (8%) patients. Of the 53 patients with recurrent reflux, 46 underwent SFJ interruption. CONCLUSIONS: elimination of reflux in the GSV after the interruption of insufficient collaterals is only temporary. PMID- 12552479 TI - Stump pressure and transcranial Doppler for predicting shunting in carotid endarterectomy. AB - OBJECTIVES: to compare stump pressure (SP) and transcranial Doppler (TCD) with neurologic monitoring during carotid endarterectomy (CEA). MATERIALS: one hundred and forty-seven CEAs performed under local anaesthesia. METHODS: neurologic monitoring and SP were performed in all cases, while mean velocity of the middle cerebral artery (mvMCA) by TCD was done in 140/147 (95%) cases. Shunts were applied in all cases on the basis of neurologic monitoring. The following haemodynamic criteria have been compared to neurologic monitoring: (a) <25 mmHg SP; (b) <50 mmHg SP; (c) < or =10 cm/s mvMCA after carotid occlusion; (d) > or =70 decrease of mvMCA after carotid occlusion. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were calculated for each haemodynamic criterion. RESULTS: shunt was used in 18/147 (12.2%) cases. With regards to <25 mmHg SP, < or =50 mmHg SP, < or =10 cm/s mvMCA after carotid occlusion, and > or =70 decrease of mvMCA after carotid occlusion, sensitivity resulted 33, 89, 80 and 80%, respectively. Specificity resulted 96, 82, 97 and 96%, respectively. Positive predictive value resulted 55, 41, 75 and 71%, respectively. Negative predictive value 91, 98, 98 and 98%, respectively. Accuracy resulted 88, 76, 89 and 94%, respectively. CONCLUSIONS: none of the haemodynamic criteria by SP and TCD resulted absolutely reliable in predicting the need for carotid shunt. PMID- 12552480 TI - Improvement of carotid blood flow after carotid endarterectomy--evaluation using intraoperative ultrasound flow measurement. AB - OBJECTIVES: to examine the relationship between the degree of extracranial internal carotid artery (ICA) stenosis and changes in the ipsilateral ICA blood flow after carotid endarterectomy (CEA). MATERIALS AND METHODS: in a prospective study we studied 51 patients with unilateral 60-99% ICA stenosis (median degree 84%, asymptomatic stenosis n = 13, symptomatic stenosis n = 38). The degree of ICA diameter stenosis was determined by ex-vivo plastination of the surgically removed atherosclerotic specimen and video-assessed planimetry. Intraoperative transit time ultrasound flow measurements of the carotid arteries were performed before and after CEA. Blood flow changes were assessed by mathematical approximations. Statistics were done by use of the Wilcoxon signed Rank test. RESULTS: common carotid artery (CCA) and ICA median blood flow increased after CEA from 370 and 130 ml/min to 450 and 282 ml/min, respectively (p <.001). The relative increase of ICA blood flow was 5% and 18% for 60-69% and 70-79% ICA stenosis (n.s.) but 70% and 247% for 80-89% and 90-99% stenosis (p <.001 each). Mathematical evaluation (fourth-polynomal function) determined a significant increase of carotid blood flow after CEA in ICA stenosis of > or =82.3%. CONCLUSIONS: in the absence of severe contralateral ICA occlusive disease a significant increase of ipsilateral ICA blood flow by CEA can be expected in patients with an ICA stenosis of > or =82.3% (linear degree of stenosis, ECST criteria). PMID- 12552481 TI - Endoluminal repair of a complex tortuous descending thoracic aortic aneurysm. PMID- 12552482 TI - Endovascular stent grafting of a penetrating ulcer in the descending thoracic aorta. PMID- 12552483 TI - Spontaneous isolated dissection of the superior mesenteric artery. PMID- 12552484 TI - Stent-grafting for a thoracic aortic aneurysm ruptured into the right pleural cavity. PMID- 12552485 TI - Endovascular treatment of a saccular aortic arch aneurysm in a patient with a patent LIMA [correction of Lima] graft. PMID- 12552486 TI - Multiple thoracic paraspinal meningeal cysts in autosomal dominant polycystic kidney disease. AB - Spinal meningeal cysts have been reported in 3 patients as an extrarenal manifestation of autosomal dominant polycystic kidney disease (ADPKD). The authors report on a fourth patient with ADPKD who was found to harbor 7 thoracic meningeal cysts, appearing as paraspinal masses on plain films. The authors provide a comprehensive radiologic description of this abnormality. PMID- 12552487 TI - Severe renal tubular acidosis in a renal transplant recipient with repeated acute rejections and chronic allograft nephropathy. AB - Renal tubular acidosis in renal transplant recipients usually is asymptomatic and subclinical. The authors report a case of severe renal tubular acidosis manifested as muscle weakness in a renal transplant recipient. The patient received a renal transplant 30 months ago and had a history of successive episodes of acute rejection during the past 2 months. On admission, arterial blood (arterial blood pH, 7.11; pco(2), 12.8 mm Hg; and bicarbonate, 4 mEq/L [4 mmol/L]) and urine gas analysis were compatible with distal renal tubular acidosis. The graft biopsy findings showed superimposed acute rejection on chronic allograft nephropathy, and immunohistochemical staining and electron microscopic findings showed the reduced immunoactivity of H(+)ATPase pump and anion exchanger 1. The patient was treated successfully with intravenous bicarbonate and oral steroid pulse therapy. This finding suggests that rejection related renal tubular acidosis should be considered a cause of severely affected metabolic acidosis in renal transplant recipients. PMID- 12552488 TI - Assessing renal function in cirrhotic patients: problems and pitfalls. AB - Assessment of glomerular filtration rate (GFR) by common creatinine-based methods potentially is very inaccurate in patients with cirrhosis. Cirrhotic patients have several underlying conditions that contribute to falsely low serum creatinine concentrations, even in the presence of moderate to severe renal impairment, and often cause creatinine-based methods to overestimate true GFR. Such underlying conditions include decreased creatinine production secondary to decreased hepatic creatine synthesis, increased tubular creatinine secretion, and decreased skeletal muscle mass. These factors all contribute to serum creatinine concentrations that often do not accurately reflect renal function. Serum creatinine level, measured creatinine clearance, and calculated creatinine clearance may all significantly overestimate GFR; the degree of GFR overestimation was a median of 95% in published studies. Until more accurate methods of estimating GFR in cirrhotic patients are adequately validated, care should be exercised in the management of these patients because of the potential for severely impaired renal function, even in the face of normal serum creatinine concentrations. PMID- 12552489 TI - Special issues in the management of patients with ESRD and HIV infection. AB - During 1985 to 1999, the percentage of hemodialysis centers providing care to patients with human immunodeficiency virus (HIV) in the United States increased from 11% to 39%, and the percentage of dialysis patients with HIV infection increased from 0.3% to 1.4%. Therefore, nephrologists are going to be confronted not only with providing dialysis care, but also with primary care issues of these patients. In this review, we discuss issues related to HIV infection and end stage renal disease, such as choice of renal replacement therapy, vascular access, anemia, vaccination, prevention of opportunistic infections, and antiretroviral therapy, that will help nephrologists not only provide optimal care, but also improve outcomes in these patients. PMID- 12552490 TI - Drug-induced Fanconi's syndrome. AB - Fanconi's syndrome (FS) was first described by Lignac in 1924. Associated with numerous varieties of inherited and acquired conditions, FS is characterized by a generalized transport defect in the proximal tubules, leading to renal losses of glucose, phosphate, calcium, uric acid, amino acids, bicarbonates, and other organic compounds. Cardinal symptoms of the syndrome are hyperaminoaciduria, glucosuria in the face of a normal serum glucose level, and phosphate wasting. Other symptoms may be associated, such as defects in bicarbonate reabsorption; renal acidification; urate reabsorption; urinary concentration; potassium conservation; reabsorption of sodium, calcium, and low-molecular-weight proteins; and secretion of p-aminohippuric acid. Acquired renal tubular defects resulting in FS also have been described in association with many exogenous agents, whether administered or accidentally ingested. This review concentrates on drug-induced FS. PMID- 12552491 TI - Impact of timing of nephrology referral and pre-ESRD care on mortality risk among new ESRD patients in the United States. AB - BACKGROUND: Recent studies have suggested that early predialysis nephrological care is associated with lower mortality; however, this hypothesis has not been tested in a population-based study. We evaluated the impact of early nephrology referral and pre-end-stage renal disease (ESRD) care on mortality risk in a national cohort of new patients starting dialysis therapy in 1996 and 1997. METHODS: Data were obtained on a subset of patients (n = 2,264; 56%) from the Dialysis Morbidity and Mortality Study Wave 2 who then were followed up for up to 2 years. Survival comparisons were made using log-rank test, then by Cox regression adjusting for demographics, comorbid medical conditions, and surrogate markers of pre-ESRD care. RESULTS: Adjusted mortality risks (relative risks [RRs]) were higher for late- (within 4 months of dialysis initiation) compared with early-referred patients at the end of 1 and 2 years of follow-up (RR, 1.68; confidence interval [CI], 1.31 to 2.15; RR, 1.23; CI, 1.02 to 1.47, respectively). Mortality risks were similarly high for the late-referred nondiabetic (RR, 2.10; CI, 1.49 to 2.94) and hemodialysis subgroups (RR, 1.72; CI, 1.25 to 2.38). Conversely, mortality risks were lower for patients who saw a nephrologist at least twice in the year before dialysis therapy initiation (RR, 0.80; CI, 0.62 to 1.03; P = 0.08] compared with those who did not. CONCLUSION: Late nephrology referral is associated with greater death risk in new patients with ESRD, and more frequent pre-ESRD care confers increased survival benefit. These findings stress the need for earlier referral of patients to nephrologists and improved pre-ESRD care for all patients approaching ESRD in the United States to improve survival. Am J Kidney Dis 41:310-318. PMID- 12552492 TI - Beneficial effects of weight loss in overweight patients with chronic proteinuric nephropathies. AB - BACKGROUND: Proteinuria is a recognized complication of obesity, but no study has investigated the effect of weight loss in overweight patients with chronic proteinuric nephropathies of different causes. METHODS: We randomly assigned (in a 2:1 ratio) 30 overweight patients (body mass index [BMI] > 27 kg/m(2)) with diabetic and nondiabetic proteinuric nephropathies to either follow a low-calorie normoproteinic diet or maintain their usual dietary intake for 5 months. The primary outcome was change in urinary protein excretion. Secondary outcomes were changes in renal function (serum creatinine level and Cockcroft-Gault creatinine clearance) and lipid profile. RESULTS: Patients in the diet group showed a significant decrease in body weight and BMI, whereas patients in the control group showed a significant increase in body weight and BMI (between-group comparison, P < 0.05). Mean weight loss in the diet group was 4.1% +/- 3%, and 14 of 20 patients in this group lost more than 3%. Proteinuria decreased by 31.2% +/ 37% in the diet group (from protein of 2.8 +/- 1.4 to 1.9 +/- 1.4 g/24 h; P < 0.005), whereas it tended to increase in the control group (between-group comparison, P < 0.05). Changes in renal function did not differ significantly between groups, although renal function remained stable in the diet group and showed significant worsening in the control group. Serum triglyceride levels remained stable in the diet group and tended to increase in the control group (between-group comparison, P < 0.05). CONCLUSION: Moderate weight loss in overweight patients with chronic proteinuric nephropathies induces a significant decrease in proteinuria. Am J Kidney Dis 41:319-327. PMID- 12552493 TI - Fractional excretion of IgG predicts renal outcome and response to therapy in primary focal segmental glomerulosclerosis: a pilot study. AB - BACKGROUND: Prolonged treatment with steroids and/or cyclophosphamide improves the prognosis of primary focal segmental glomerulosclerosis (FSGS). In nephrotic patients, no clinical or histological feature predicts responsiveness to therapy. METHODS: In 50 patients with FSGS, fractional excretion (FE) of immunoglobulin G (IgG), albumin, transferrin, and alpha(1)-microglobulin (alpha(1)m) was calculated. The aim of the study is to assess whether FE IgG and FE alpha(1)m: (1) correlate with histological lesions, (2) predict outcome, and (3) may be useful to guide therapy. RESULTS: The association of FE IgG with percentage of glomeruli with segmental sclerosis was at the limit of significance (P = 0.01). FE alpha(1)m was associated with extent of tubulointerstitial damage (P = 0.008). By multiple regression analysis, FE alpha(1)m was dependent on FE IgG (R(2) = 0.76; P = 0.000). The predictive value of proteinuric variables on outcome was evaluated in 29 patients with nephrotic syndrome and baseline normal renal function (serum creatinine level, 1.04 +/- 0.22 mg/dL [92 +/- 19 micromol/L]; follow-up, 50 +/- 33 months); remission rates were 91% and 0% in patients with FE IgG less than versus greater than 0.140 (P = 0.0009). By multiple logistic regression analysis, only FE IgG was associated with remission (P = 0.043). Proteinuria less than versus greater than 7.5 g/d of protein predicted end-stage renal failure (0% versus 36%; P = 0.004); the predictive value of FE IgG less than versus greater than 0.140 was higher (0% versus 71%; P = 0.0000). Patients with FE IgG less than 0.025 were responsive to steroids alone (70%) or steroids and cyclophosphamide (20%); patients with FE IgG greater than 0.025 and less than 0.140 were responsive to steroids alone (20%) or steroids and cyclophosphamide (80%); and 100% of patients with FE IgG greater than 0.140 were unresponsive to therapy (P = 0.000). CONCLUSION: In FSGS, FE IgG is at the limit of statistically significant association with segmental sclerosis, and FE alpha(1)m is associated with extent of tubulointerstitial damage. FE IgG shows the best predictive value for remission, progression, and response to therapy and may be useful to guide treatment. Am J Kidney Dis 41:328-335. PMID- 12552494 TI - High prevalence of immuno-unreactive intact albumin in urine of diabetic patients. AB - BACKGROUND: Intact albumin in urine may exist in two forms, immunoreactive and immuno-unreactive. Previous estimates of albuminuria in diabetic urine have only detected immunoreactive forms. METHODS: High performance liquid chromatography (HPLC) was used in this study to measure both forms of intact albumin (termed total intact albumin) to provide a more accurate measurement of albuminuria compared with radioimmunoassay (RIA) on 97 fresh urine samples from patients with diabetes. Eighty-six control urine samples from volunteers without diabetes were also tested. RESULTS: There was no significant difference between the two methods for nondiabetic controls. For diabetic urine samples, 91.6% of samples showed a greater concentration of albumin measured by HPLC than RIA. For normoalbuminuric diabetic samples, HPLC gave a mean albumin excretion rate of 12.5 +/- 4.4 microgram/min (SD), whereas RIA gave a rate of 8.0 +/- 6.7 microgram/min (P = 0.004; N = 28). For microalbuminuric samples, there also was a statistically significant difference: HPLC albumin excretion rate, 82.0 +/- 49.9 microgram/min, and RIA, 49.0 +/- 34.6 microgram/min (P = 0.004; N = 30). Thirty-two urine samples were normoalbuminuric by RIA (albumin, 11.4 +/- 3.9 microgram/min), but in the microalbuminuric range as determined by HPLC (albumin, 38.5 +/- 14.4 microgram/min). For urine samples in the macroalbuminuric range, there was no statistically significant difference between HPLC and RIA. Immuno-unreactive albumin was confirmed as albumin, analyzed by two-dimensional electrophoresis and matrix-assisted laser desorption ionization mass spectrometry. CONCLUSION: These studies show that to determine microalbuminuria accurately, there is a need to assess urinary total intact albumin, rather than simply immunoreactive albumin. Am J Kidney Dis 41:336-342. PMID- 12552495 TI - IgM nephropathy: clinical picture and long-term prognosis. AB - BACKGROUND: Immunoglobulin M (IgM) nephropathy is an idiopathic glomerulonephritis with mesangial hypercellularity and diffuse IgM deposits. METHODS: We studied clinical presentation, morphological findings, and prognostic factors in 110 patients with IgM nephropathy without systemic diseases. The series included both pediatric and adult patients with nephrotic syndrome (NS) or minor urinary abnormalities. RESULTS: Mean postbiopsy follow-up was 8 years. During 15 years of follow-up, 36% of patients developed renal insufficiency and 23% reached end-stage renal failure. In multivariate analysis, hypertension at the time of renal biopsy was the only significant risk factor for renal insufficiency. Of histological parameters, interstitial fibrosis had the strongest prognostic value. Hypertension was diagnosed in 50% of patients with a postbiopsy follow-up of 15 years. Twenty-nine percent of nephrotic patients had disease resistant to corticosteroids, whereas 80% of patients with steroid sensitive disease were steroid dependent. Eleven patients, 8 patients with NS and 3 patients with asymptomatic proteinuria, underwent repeated renal biopsy. In five samples, typical morphological characteristics of focal and segmental glomerulosclerosis were seen. CONCLUSION: We propose that IgM nephropathy can be divided into two subgroups with similar renal biopsy findings, but differences in sex distribution and initial presentation. PMID- 12552496 TI - Renal artery stenosis by three-dimensional magnetic resonance angiography in type 2 diabetics with uncontrolled hypertension and chronic renal insufficiency: prevalence and effect on renal function. AB - BACKGROUND: The variable course of renal disease in type 2 diabetes mellitus in part may reflect associated atherosclerotic nephropathy. METHODS: To determine the influence of subcritical (<65%) renal artery stenosis (RAS) on the progression of chronic kidney disease, 45 patients with type 2 diabetes with uncontrolled hypertension and serum creatinine levels of 1.8 mg/dL or greater (>/=159.1 micromol/L) were screened by three-dimensional magnetic resonance angiography (MRA). Mean monthly decrease in reciprocal serum creatinine x 100 and time to initiation of dialysis therapy, adjusting for baseline serum creatinine level, were compared in those with and without RAS. Follow-up was censored at the time of death or angioplasty. RESULTS: At baseline, RAS-negative (RAS(-); n = 27) and RAS-positive (RAS(+); n = 18) groups were similar in duration of diabetes and hypertension, hyperlipidemia, blood pressure, diabetic management, and renal function. RAS(+) subjects were older (P = 0.04) and more likely to have claudication (P = 0.006), smoke (P = 0.02), and have heart disease (P = 0.06). During a median follow-up of 9.4 months, 3 patients underwent stent placement, 2 patients died, and 12 patients progressed to dialysis therapy. The RAS(+) group had a more rapid monthly decline in reciprocal serum creatinine x 100 (mean, 1.63 +/- 0.9 versus 0.69 +/- 1.0 [SD]; P = 0.04). The relative risk for progression to end-stage renal disease was 2.4 in the RAS(+) versus RAS(-) group. Multivariate analysis showed that this effect was not independent of several established atherosclerotic risk factors. CONCLUSION: MRA-detected RAS is common (40%) in patients with type 2 diabetes with uncontrolled hypertension and renal insufficiency. Subcritical (<65%) RAS is a significant risk factor for progressive renal failure. PMID- 12552497 TI - Platelet endothelial cell adhesion molecule-1 and angiogenic factor expression in idiopathic membranous nephropathy. AB - BACKGROUND: Idiopathic membranous nephropathy (IMN), a principal disease of glomerular capillaries, was investigated for some aspects of glomerular capillary injury and repair (angiogenesis). METHODS: Fifteen cases of IMN were studied immunohistochemically for expression of the endothelial cell antigen platelet endothelial cell adhesion molecule-1 (PECAM-1[CD31]) and the angiogenesis stimulating factors vascular endothelial growth factor (VEGF) and thymidine phosphorylase (TP). An equal number of normal control kidneys of fetal and mature origin were tested for the same antigens. RESULTS: Normal tissues expressed PECAM 1 in both glomerular and interstitial endothelial cells, whereas VEGF and TP were expressed in the tubular epithelium. IMN was characterized by complete or partial loss of PECAM-1 expression from glomerular capillaries and a parallel gain/expression of this antigen by the tubular epithelium. In addition, VEGF and TP expression was lost or considerably reduced from tubular cells of IMN. CONCLUSION: We hypothesize that PECAM-1 expression by tubular epithelial cells represents uptake of CD31(+) cell-surface fragments released by glomerular endothelial cells after glomerular damage. The damage is confounded by the failure of angiogenic mechanisms to promote glomerular angiogenesis (repair) because both VEGF and TP stimulation by the tubular epithelium is eliminated. It is suggested that immunohistochemical detection of VEGF or TP in the tubular epithelium may be useful in understanding the pathogenesis of IMN. PMID- 12552498 TI - Group A streptococcal antigen in the glomeruli of children with Henoch-Schonlein nephritis. AB - BACKGROUND: Although the pathogenesis of Henoch-Schonlein nephritis (HSN) remains unclear, there is substantial evidence that it is an immune complex-mediated disease. HSN is preceded by upper-respiratory tract infection in 30% to 50% of patients, but there is no evidence that group A streptococcal (GAS) infection has a pathogenetic role in this disease. Recently, nephritis-associated plasmin receptor (NAPlr), a GAS antigen, was found primarily in the glomerular mesangium of patients with early-stage acute poststreptococcal glomerulonephritis. METHODS: To determine the possible role of NAPlr in HSN, expression of the receptor was determined in glomeruli using fluorescein isothiocyanate-labeled rabbit polyclonal anti-NAPIr antibody, and serum antistreptolysin O (ASO) titers were measured in children with HSN. RESULTS: Ten of 33 patients (30%) with HSN showed segmental or global mesangial staining with NAPlr antibody, whereas only 4 of 120 patients (3%) with other renal diseases were positive (P < 0.001, Fisher's exact test). Patients with HSN also showed significantly greater ASO titers than patients with other renal diseases (P = 0.03, Mann-Whitney U test). Serum ASO titers were significantly greater in patients with HSN with than without glomerular NAPlr antigen (P = 0.03, Mann-Whitney U test). CONCLUSION: These findings suggest that the deposition of NAPlr in the mesangium, induced by GAS infection, may have a role in the pathogenesis of HSN in some patients. Am J Kidney Dis 41:366-370. PMID- 12552499 TI - Impact of interferon-gamma and interleukin-4 gene polymorphisms on development and progression of IgA nephropathy in Japanese patients. AB - BACKGROUND: Cytokines have an important role in the pathogenesis and disease progression of immunoglobulin A (IgA) nephropathy. The aim of this study is to investigate the impact of gene polymorphisms of T helper cell subtype 1 (T(H)1)/T(H)2 cytokines, interferon-gamma (IFN-gamma), and interleukin-4 (IL-4) on IgA nephropathy in Japanese patients. METHODS: We investigated IFN-gamma gene (IFNG) and IL-4 gene (IL4) polymorphisms in 96 patients with biopsy-confirmed IgA nephropathy who were followed-up for more than 3 years in our outpatient clinic and 61 healthy controls by polymerase chain reaction and direct sequencing methods. IFNG polymorphism was characterized as a microsatellite of intron 1. Four alleles were identified and designated IFNG 112, 114, 116, and 118, corresponding to 12, 13, 14, and 15 repeats, respectively. A variable number of tandem repeat (VNTR) polymorphisms of IL4 also were studied, and alleles were designated IL4 B1 and B2, corresponding to 2 and 3 repeats, respectively. RESULTS: In patients with IgA nephropathy, IFNG 114 allele and IFNG 114(+/+) genotype frequencies were significantly greater than in the healthy control group (60% versus 45%; P < 0.01 and 43% versus 23%; P < 0.05, respectively), but there was no difference between the IgA nephropathy and healthy control groups in frequencies of both IL4 VNTR allele and genotype. However, frequencies of IL4 B1 allele and B1/B1 genotype in patients with progressive IgA nephropathy (end-stage renal disease or doubling of serum creatinine level; n = 34) were significantly greater than corresponding values in the nonprogression group (n = 62; 79% versus 61%; P < 0.01 and 59% versus 34%; P < 0.05, respectively). We could not confirm an association between IgA nephropathy and polymorphisms of genes involved in the renin-angiotensin system. CONCLUSION: Our results suggest that IFN-gamma and IL-4 gene polymorphisms could influence disease susceptibility and disease progression in IgA nephropathy in Japanese patients. Am J Kidney Dis 41:371-379. PMID- 12552500 TI - Why patients with ESRD do not select self-care dialysis as a treatment option. AB - BACKGROUND: In-center hemodialysis is the most prevalent (and resource-intense) form of dialysis in North America despite many patients being capable of performing dialysis themselves. The purpose of this study is to describe reasons in-center hemodialysis patients choose not to perform self-care dialysis and identify variables associated with a negative attitude toward self-care dialysis. METHODS: We conducted a cross-sectional survey (return rate, 83%) of prevalent in center hemodialysis patients and combined this with demographic and comorbidity data obtained from our prospectively maintained database. We also performed multiple logistic regression to determine factors associated with the attitude, "patients should not perform dialysis without being supervised by a nurse." RESULTS: The most prevalent knowledge barrier was lack of a satisfactory explanation of the various techniques. The most prevalent attitude barriers were that patients should not dialyze without direct supervision, fear of failure to perform self-care dialysis adequately, and fear of social isolation. The most prevalent skill barriers were needle phobia and lack of space at home. Variables significantly associated with a negative attitude toward self-care dialysis were age/fear of substandard care (interaction), needle phobia, fear of change, fear of social isolation, and unwillingness to remain awake during dialysis. CONCLUSION: This study identified a variety of barriers to self-care dialysis, and these results are being used to direct changes to our program aimed at increasing the uptake of self-care hemodialysis, which we believe will benefit both patients and health care providers and may offer a solution to nursing and resource shortages. PMID- 12552501 TI - Factors associated with medication-related problems in ambulatory hemodialysis patients. AB - BACKGROUND: Hemodialysis (HD) patients are at risk for medication-related problems. Patient characteristics associated with the number of medication related problems in HD patients have not been investigated. METHODS: Patient records were reviewed to identify medical problems, prescribed medications, medication indication(s), and medication-related problems. Medication classes and medication-related problems were compared between patients with and without diabetes mellitus (DM). Correlations were performed to determine whether associations exist between medication-related problems, number of medications, number of medication doses per day, number of comorbid conditions, patient age, and duration of end-stage renal disease while controlling for DM status. RESULTS: Medical records of 133 patients were evaluated. Patients were 60.5 +/- 15.2 years old, prescribed 11.0 +/- 4.2 medications, and had 6.0 +/- 2.3 comorbidities. Medication-related problems were identified in 97.7% of patients. Four hundred seventy-five medication-related problems were identified, averaging 3.6 +/- 1.8 medication-related problems per patient. Patients with DM had more medication related problems identified than those without DM (303 versus 172 medication related problems, respectively; P < 0.05). Medication-related problems correlated positively with number of patient comorbidities (P < 0.001). CONCLUSION: Medication-related problems are prevalent in virtually all HD patients. The number of medication-related problems in an individual patient increases as the number of comorbid conditions increases. The most frequent medication-related problems were drug without indication (30.9%), laboratory (27.6%), indication without drug use (17.5%), and dosing errors (15.4%). Patients with DM are at increased risk for medication-related problems. Health care providers taking care of HD patients should be aware of this problem, and efforts to avoid or resolve medication-related problems should be undertaken at all HD clinics. PMID- 12552502 TI - Daytime sleepiness in stable hemodialysis patients. AB - BACKGROUND: Patients frequently sleep during hemodialysis (HD), a behavior often attributed to treatment-related fatigue and/or simple boredom. The possibility that this behavior reflects a more pervasive underlying increase in daytime sleepiness has never been systematically examined. Thus, we studied a sample of HD patients on an off-dialysis day to establish the presence or absence of daytime sleepiness independent of effects of treatment, quantify its severity, and identify associated demographic, metabolic, and sleep-related variables. METHODS: Forty-six stable HD patients underwent polysomnography, followed the next day (a nondialysis day) by the Multiple Sleep Latency Test (MSLT; low score = greater sleepiness), a measure of physiological daytime sleepiness. Subjects also completed the Epworth Sleepiness Scale (ESS; high score = greater sleepiness), a measure of subjective daytime sleepiness. RESULTS: One third (n = 15) of subjects had MSLT scores suggesting abnormal levels of physiological daytime sleepiness, and six subjects had scores consistent with severe, pathological sleepiness. Thirty percent (n = 14) had significant subjective daytime sleepiness as measured by the ESS. However, MSLT and ESS scores were unrelated. Higher indices of sleep apnea (r = -0.324; P = 0.028) and brief arousals (r = -0.370; P = 0.009) correlated significantly with increased physiological, but not subjective, sleepiness. Longer nocturnal sleep latencies (r = 0.350; P = 0.017) and greater percentage of rapid-eye-movement sleep (r = 0.302; P = 0.042) were associated with decreased physiological sleepiness. Other major demographic, metabolic, and sleep-related variables did not correlate with MSLT scores, and none of the variables examined were related to ESS scores. CONCLUSION: Daytime sleepiness is common in HD patients and may be severe despite the absence of obvious clinical risk factors for the condition. Thus, research designed to identify cost-effective indicators of daytime sleepiness and evaluate the detrimental effects of sleepiness on clinical outcomes in HD patients is warranted. PMID- 12552503 TI - Daytime sleepiness in patients with CRF: impact of nocturnal hemodialysis. AB - BACKGROUND: Patients with end-stage renal disease (ESRD) have a high prevalence of sleep disorders, which are not improved by conventional hemodialysis (CHD). Although sleep disorders are commonly associated with complaints of excessive daytime sleepiness, the severity and pathogenesis of daytime sleepiness has not been evaluated objectively in patients with ESRD. Nocturnal hemodialysis (NHD) is a new technique that provides better clearance of uremic toxins than CHD and, consequently, may improve sleep quality and daytime sleepiness. The authors wished to determine the severity and pathogenesis of daytime sleepiness in patients with ESRD and evaluate the impact of NHD. METHODS: Sleep quality was monitored by overnight polysomnography, and daytime sleepiness was assessed by the multiple sleep latency test (MSLT). These measurements were performed in 24 patients (15 men and 9 women, 44 +/- 10 years) while on treatment with CHD and were repeated in 15 patients after conversion to NHD. RESULTS: The majority (54%) of patients on CHD were pathologically sleepy (somnolent group, mean sleep latency <5 minutes), and, in comparison with the remaining patients (alert group, mean sleep latency >5 minutes), their blood urea nitrogen (BUN; 77.9 +/- 9.8 v 60.2 +/- 12.0 mg/dL, P < 0.001; 27.8 +/- 3.5 v 21.5 +/- 4.3 mmol/L; P < 0.001), and periodic limb movement (PLM) index (57 +/- 47 v 6 +/- 10/hr; P = 0.002) were significantly higher. Furthermore, sleep latency was correlated with BUN (R = 0.58, P = 0.008). After conversion to NHD, there was a significant fall in BUN and the severity of sleep apnea, but the overall frequency of PLM and sleep fragmentation remained elevated. Nevertheless, there was a trend for the Somnolent group to become less sleepy on NHD, and this was associated with a modest reduction in the frequency of PLM. CONCLUSION: Excessive daytime sleepiness occurs in approximately 50% of patients with ESRD. The etiology appears to be related both to uremia and sleep fragmentation associated with PLM. PMID- 12552504 TI - The relationship between serum calcium and phosphate levels and cardiac valvular procedures in the hemodialysis population. AB - BACKGROUND: Cardiac valvular disease is a common complication in hemodialysis patients, with a prevalence of up to 9%. In these patients, calcium-phosphate imbalance is associated with systemic and cardiac calcification. We investigated the relationship between abnormal calcium and phosphate levels and valvular disease sufficiently severe to require an invasive procedure. METHODS: We analyzed data from 12,509 hemodialysis patients from the US Renal Data System database, 204 of whom underwent a valvular procedure. All were prevalent in center dialysis patients as of 1993, when cross-sectional data were collected. RESULTS: In a Cox multivariate model, a serum phosphate level of 5.0 mg/dL or greater (>/=1.62 mmol/L) was associated with increased risk for a valvular procedure compared with a phosphate level less than 5 mg/dL (<1.62 mmol/L; hazard ratio, 1.47; P = 0.033). A calcium level less than 8.8 mg/dL (<2.2 mmol/L) was associated with fewer valvular procedures compared with a normal calcium level (hazard ratio, 0.61; P = 0.018). However, a high calcium level (>10.5 mg/dL [>2.63 mmol/L]) had no significant relationship with the outcome (hazard ratio, 0.89; P = 0.65) compared with a normal level. Calcium-phosphate product was not significant as an interaction term and therefore was not included in the final analysis. The relationship of parathyroid hormone (PTH) level to outcome was not significant. CONCLUSION: A serum phosphate level of 5.0 mg/dL or greater (>/=1.62 mmol/L) is associated with increased risk for a valvular procedure, and a low calcium level is associated with fewer valvular procedures. There is no compelling evidence that elevated calcium or PTH levels have a significant relationship to valvular disease that results in an invasive procedure. PMID- 12552505 TI - Preexisting intimal hyperplasia of radial artery is associated with early failure of radiocephalic arteriovenous fistula in hemodialysis patients. AB - BACKGROUND: The radiocephalic arteriovenous fistula (AVF), which provides the best vascular access for hemodialysis, continues to have a high incidence of early failure. Intimal hyperplasia (IH) of the radial artery is observed commonly in uremic patients before hemodialysis, but the impact of this preexisting IH on the early failure of radiocephalic AVFs has not been reported yet. Therefore, we designed this study to: (1) investigate clinical risk factors for IH, and (2) determine whether preexisting IH of the radial artery is associated with early failure of a radiocephalic AVF. METHODS: Specimens from the radial artery were obtained during the radiocephalic AVF operation. IH was measured with trichrome staining, and AVF patency was prospectively followed up for 12 months after the operation. RESULTS: Of the 59 patients, 45 patients had evidence of IH in their radial artery (76.2%). Patients with IH (n = 45) were older than those without IH (n = 14; 58 +/- 12 versus 44 +/- 17 years; P = 0.003). The incidence of diabetes mellitus in patients with IH was greater than that in patients without IH (60.0% versus 28.6%; P = 0.004). Of the 57 patients, except for 2 patients who died before the end point of the study with patent AVFs, fistula failure was observed only in patients with IH (22 of 44 patients; 50% versus 0%; P < 0.001). The intima was thicker in the failed-AVF group than the patent-AVF group (93.1 +/- 37.5 versus 45.6 +/- 17.4 micrometer P < 0.001). CONCLUSION: This study suggests that early failure of radiocephalic AVFs in hemodialysis patients is closely associated with preexisting IH of the radial artery. PMID- 12552506 TI - Venous access: women are equal. AB - BACKGROUND: Arteriovenous fistulae (AVFs) are the preferred method of vascular access for hemodialysis patients; however, the current rate of AVF placement is only 25% to 30% in the United States. This prevalence is even smaller among women and attributed to their perceived inadequate vasculature. This commonly held view that women have less adequate vasculature for AVF placement than men has not been shown objectively in the literature. METHODS: To determine a difference in vasculature between the sexes, we retrospectively analyzed data on preoperative vascular mapping in 192 patients. During a 2-year period, vascular mapping of the upper extremities was routinely performed using duplex ultrasound in all patients requiring vascular access in our institution. RESULTS: One hundred six of these 192 patients were women, and 86 were men. There was no significant difference in vein size between men and women at any of the sites measured. A total of 87 fistulae were placed in 140 patients, 49% in women and 51% in men (P = 0.16). Prevalences of AVFs were 57% and 68% (P = 0.2) in women and men, respectively. Percentages of fistulae used at the initiation of hemodialysis therapy were 72% in women and 77% in men (P = 0.57). CONCLUSION: These data support the view that women have adequate vasculature for the placement of AVFs compared with men. We cannot explain the different outcomes found in other studies; however, we suggest that the differences are caused by physician practice patterns, not anatomic differences between men and women. PMID- 12552507 TI - Comparison of urea reduction ratio and hematocrit data reported in different data systems: results from the Centers for Medicare & Medicaid Services and the Renal Network Inc. AB - BACKGROUND: Urea reduction ratio (URR) and hematocrit values reported on the Centers for Medicare & Medicaid Services (CMS) claims were compared with data from two different databases. METHODS: URRs and hematocrits from two different CMS databases (National Claims History and End-Stage Renal Disease Clinical Performance Measures [CPM] Project) and one Network database (The Renal Network Data System [TRNDS]) were compared for October through December 1998 and December 1998, respectively. A sample of records from the regional database was validated by independent chart review. RESULTS: Nationally, the percentage of agreement for patients with URRs of 65% or greater and less than 65% was 94% (kappa, 0.81; 95% confidence interval [CI], 0.80 to 0.83); regionally, the percentage of agreement was 95% (kappa, 0.85; 95% CI, 0.84 to 0.86). Nationally, linear regression of hematocrit values from both data sources yielded r(2) congruent with 0.61 each month and r(2) = 0.70 for average values during the 3-month study period. Nationally, the percentage of agreement for patients with hematocrits of 33% or greater and less than 33% was 84% (kappa approximately 0.66) each month. Regionally, linear regression of monthly hematocrit values from both data sources yielded r(2) = 0.66, and percentage of agreement for patients with hematocrits of 33% or greater and less than 33% was 87% (kappa, 0.71; 95% CI, 0.70 to 0.73). Validation of a sample of records in the TRNDS database resulted in 98% agreement for patients with URRs of 65% or greater and less than 65% and 96% agreement for patients with hematocrits of 33% or greater and less than 33%. CONCLUSION: Although there is general agreement between clinical variables submitted on the claims and in the CPM Project, some variation exists. Data from either source yield the same information when classifying patients as above or below threshold values. PMID- 12552508 TI - The effect of N-acetylcysteine on plasma total homocysteine levels in hemodialysis: a randomized, controlled study. AB - BACKGROUND: The chronic hemodialysis population has an accelerated rate of cardiovascular morbidity and death. Furthermore, elevated levels of the putative atherothrombotic risk factor homocysteine are almost ubiquitous in this population. Attempts to normalize elevated plasma total homocysteine (tHcy) levels in dialysis patients using pharmacological-dose vitamin therapy or other strategies generally have been unsuccessful. Preliminary uncontrolled evidence suggests that N-acetylcysteine (NAC) may be an effective tHcy-lowering agent. We designed a randomized placebo-controlled study to determine the effect of prolonged oral NAC therapy on lowering tHcy levels in vitamin-replete chronic hemodialysis patients. METHODS: Thirty-eight subjects were treated before intervention with a standard dialysis vitamin supplement to ensure a uniform vitamin-replete state. They were then block randomized to treatment with NAC, 1.2 g twice a day, for 4 weeks or matched placebo. RESULTS: There were no significant baseline differences between the two groups, although differences in pyridoxal 5' phosphate (active form of vitamin B(6)) levels approached significance (P = 0.06). In a paired analysis, there was no statistically significant difference between the NAC and placebo groups. NAC was very well tolerated in hemodialysis patients. CONCLUSION: This randomized placebo-controlled trial found that chronic oral NAC therapy did not significantly reduce tHcy levels in hemodialysis patients. Although a larger sample size theoretically could have increased the statistical significance between groups, implications of the potentially very modest reduction in tHcy levels are not yet known. Finally, based on this limited study, NAC appears to be a safe and well-tolerated therapy in the hemodialysis population. PMID- 12552509 TI - Decreased survival among sedentary patients undergoing dialysis: results from the dialysis morbidity and mortality study wave 2. AB - BACKGROUND: Sedentary behavior is associated with an increased risk for death in the general population. However, the association between inactivity and mortality has not been studied in a large cohort of dialysis patients despite the high prevalence of sedentary behavior in this group. METHODS: We used the Dialysis Morbidity and Mortality Study Wave 2, a prospective study of a national sample of 4,024 incident peritoneal dialysis and hemodialysis patients from 1996 to 1997, to determine whether sedentary behavior is associated with increased mortality during a 1-year period in this group after adjusting for confounding variables. RESULTS: The study population consisted of the 2,837 patients with accurate survival data who were able to ambulate and transfer. Eleven percent of the sedentary patients died during the study period compared with 5% of nonsedentary patients. In a survival analysis, sedentary behavior (hazard ratio, 1.62; 95% confidence interval, 1.16 to 2.27) was associated with an increased risk for death at 1 year after adjusting for all variables that we postulated might be associated with survival and for differences between sedentary and nonsedentary patients. CONCLUSION: Sedentary behavior is associated with an increased risk for mortality among dialysis patients similar in magnitude to that of other well established risk factors, such as a one-point reduction in serum albumin concentration. More attention should be given to exercise behavior in dialysis patients, and controlled clinical trials are needed to further define the association of sedentary behavior with mortality. PMID- 12552510 TI - Renal transplantation in children with lupus nephritis. AB - BACKGROUND: This study was conducted to determine renal allograft and patient survival for children with lupus nephritis compared with pediatric allograft recipients with other causes of end-stage renal disease. METHODS: Data from the United Network for Organ Sharing registry for children aged 1 to 21 years who received an initial renal transplant between 1987 and 1997 were included. Subjects with kidney failure caused by other vasculitides were excluded, as were those with multiple organ transplants. RESULTS: A total of 7,926 children (n = 254 children with lupus) were available for evaluation. Patients with lupus were older (median age, 19 versus 15 years; P < 0.01) and more commonly female (80% versus 40%; P < 0.01) and black (34% versus 20%; P < 0.01) than patients without lupus. Median follow-up was 4.2 years (range, 0.03 to 13 years). Renal allograft survival was not different between groups when adjusting for sex, race, age, and cadaveric (CAD) transplant (P = 0.98). Among patients with lupus, those with CAD allografts were 1.9 times (95% confidence interval [CI], 1.1 to 3.3; P = 0.02) more likely to lose their graft than those with living donor grafts. This increased risk for graft failure with CAD transplants was observed to a lesser degree in subjects without lupus (hazard ratio, 1.3; 95% CI, 1.2 to 1.5). Patients with lupus were 1.8 times more likely to die compared with patients without lupus in multivariate analysis (95% CI, 1.14 to 2.74; P = 0.01). CONCLUSION: Although allograft survival is not different, patient survival after renal transplantation is less for pediatric patients with lupus compared with other pediatric renal allograft recipients. PMID- 12552511 TI - Long-term survival following simultaneous kidney-pancreas transplantation versus kidney transplantation alone in patients with type 1 diabetes mellitus and renal failure. AB - BACKGROUND: Pancreas transplantation improves quality of life and prevents the progression of secondary complications of diabetes. Whether these benefits translate into a long-term survival advantage is not entirely clear. METHODS: Using the United Network for Organ Sharing database, we analyzed long-term survival in 18,549 patients with type 1 diabetes and renal failure who received a kidney transplant between 1987 and 1996. Patient survival was calculated using the Kaplan-Meier method. Proportional hazards models were used to adjust for effects of differences in recipient and donor variables between simultaneous kidney-pancreas transplants (SKPTs) and kidney-alone transplants. RESULTS: SKPT and living donor kidney recipients had a significant crude survival distribution advantage over cadaver kidney transplant recipients (8-year survival rates: 72% for SKPT recipients, 72% for living donor kidney recipients, and 55% for cadaver kidney recipients). The survival advantage for SKPT recipients over cadaver kidney recipients diminished, but persisted after adjusting for donor and recipient variables and kidney graft function as time-varying covariates. SKPT recipients had a high mortality risk relative to living donor kidney recipients through 18 months posttransplantation (hazards ratio, 2.2; P < 0.001), but had a lower relative risk (hazard ratio, 0.86; P < 0.02) thereafter. In SKPT recipients, maintenance of a functioning pancreas graft was associated with a survival benefit. CONCLUSION: The long-term survival of SKPT recipients is superior to that of cadaver kidney transplant recipients with type 1 diabetes. There is no difference in survival of SKPT recipients and living donor kidney recipients with type 1 diabetes at up to 8 years' follow-up; the former have a greater early mortality risk and the latter have a greater late mortality risk. Results of this study suggest that successful simultaneous kidney-pancreas transplantation is not only life enhancing, but life saving. PMID- 12552512 TI - De novo thrombotic microangiopathy in renal transplant recipients: a comparison of hemolytic uremic syndrome with localized renal thrombotic microangiopathy. AB - BACKGROUND: Thrombotic microangiopathy (TMA) is a well-recognized and serious complication of renal transplantation, affecting 3% to 14% of patients administered calcineurin-inhibitor-based immunosuppression. METHODS: We reviewed 1,219 biopsy reports of 742 kidney and kidney-pancreas transplants performed during 15 years at our center and found 21 biopsy-confirmed cases of TMA. RESULTS: On presentation, the majority (62%) had systemic TMA with manifest hemolysis and thrombocytopenia, whereas a subset had TMA localized only to the graft (38%). There were no statistically significant differences in sex, type of transplant, age, race, or type of immunosuppression. Patients with systemic TMA were more likely to be treated with plasma exchange (38% versus 13%; P < 0.05), more often required dialysis therapy (54% versus 0%; P = 0.01), and had a greater rate of graft loss (38% versus 0%; P < 0.05). No patient with the localized variant had TMA-related graft loss. Patients with localized TMA often responded to reduction, conversion, or temporary discontinuation of calcineurin-inhibitor based immunosuppression therapy and did not routinely require plasma exchange for graft salvage. We compare our findings with the literature regarding the prognosis of TMA. CONCLUSION: Classifying patients with post-renal transplantation TMA into those with localized and systemic disease is clinically useful because each group has distinct characteristics and clinical courses. PMID- 12552513 TI - Demographics and trends in overweight and obesity in patients at time of kidney transplantation. AB - BACKGROUND: The epidemic of overweight and obesity is one of the most serious public health problems in the United States, although information regarding its effect on chronic kidney transplant patients is sparse. The authors describe the prevalence, demographics, and trends in overweight and obesity at the time of kidney transplantation. METHODS: Data were obtained from a national transplant database that included all kidney transplants since 1987. Body mass indices (BMI) at time of transplantation were stratified by demographic categories and year. Trends of overweight and obesity between the general population and subjects undergoing kidney transplantation were compared. Logistic regression analyses were performed to identify variables associated with increased likelihood of being obese. RESULTS: The majority (60%) of subjects at time of transplantation currently are overweight or obese. Between 1987 and 2001, the proportion of obese transplant recipients rose by 116%. The rate of increase was grossly similar to that in the general population. The likelihood of being obese increased with age, female sex, noninsulin-dependent diabetes mellitus, black race, and the more recent the transplant year. Conversely, the proportion of recipients with lower BMI fell by approximately 50%. CONCLUSIONS: Overweight and obesity are very prevalent at the time of kidney transplantation and are eclipsing protein-energy malnutrition as the more common nutritional illness. This may have profound negative effects on patient and allograft outcomes. Nephrologists may need to reexamine traditional notions of nutritional disease and therapeutic strategies to more effectively deal with this formidable challenge. PMID- 12552514 TI - Pseudoaneurysm of the iliac artery secondary to Aspergillus infection in two recipients of kidney transplants from the same donor. AB - The authors report 2 cases of patients who underwent cadaveric renal transplantation from the same donor in a multiorgan extraction procedure. Both cases showed, during the first 6 months posttransplantation, a worsening in renal graft function and signs of ischemia in the homolateral lower limb. One of the cases was preceded by pain in the sciatic region. In imaging tests, a pseudoaneurysm was detected in the iliac artery in both patients. Grafts had to be removed, and the iliac arteries were ligated with posterior isolation of Aspergillus spp from the arterial vessels but not from the renal tissue. Besides surgery, medical treatment with liposomal amphotericin B was initiated with a different outcome in each patient: patient A died, whereas patient B recovered. The absence of Aspergillus spp infection in liver and heart recipients ruled out a donor-transmitted infection. The graft placements were carried out in different operating rooms, which rules out contamination during the transplantation process. All of this leads us to conclude that the infection must have occurred during the preservation phase of the kidney. PMID- 12552515 TI - Drug interaction in a renal transplant patient: cyclosporin-Neoral and orlistat. AB - An overweight 56-year-old type II diabetic on peritoneal dialysis (body mass index 35 kg/m(2)) was taking Orlistat for some months up until live-unrelated renal transplantation. Despite oral cyclosporin A (CyA) for 48 hours pretransplantation, it was very difficult to achieve adequate CyA blood levels for the first week postengraftment despite the use of much larger oral CyA doses. After opening his bowels on day 7, and the use of 3 days intravenous CyA, good CyA blood levels were achieved then maintained with conventional oral doses. The authors believe that this case shows important interactions between CyA and Orlistat. PMID- 12552516 TI - A monoclonal V kappa l light chain responsible for incomplete proximal tubulopathy. AB - Calcium and phosphate metabolism abnormalities are frequent in myeloma patients and the role of renal lesions in such ionic perturbations may have been overlooked. The authors herein report the complete primary structure of a Bence Jones Vkappal light chain responsible for myeloma-associated proximal tubulopathy with increased phosphaturia. Plasma and serum biochemical evaluations indicated a proximal tubular dysfunction mainly manifested as tubular acidosis and phosphate loss. The study of a kidney biopsy showed interstitial and tubular lesions with numerous myeloma casts and peculiar features of the proximal tubular cells, which carried numerous phagolysosomal inclusions with occasional crystalline periodic striation. The nephrotoxic light chain primary structure was deduced from the bone marrow monoclonal plasma cells RNA. The kappal sequence was highly homologous to kappa chains previously characterized in patients with Fanconi syndrome. It was related to the Vkappal subgroup and was composed of a variable segment encoded by the O8/O18 germline gene rearranged to Jkappa4. The primary sequence presented unusual features restricted to the variable region, including substitutions of residues 28 and 31 in the complementary determining region 1 (CDR1) by amino acids of different charge. An unusual conformation of the kappal domain, likely resulting from somatic hypermutation, could alter the catabolism of the protein after its internalization and result in the tubular cell dysfunction. Comparison with Fanconi syndrome studies suggests that Vkappal Bence Jones proteins may damage proximal tubular cells to an extent varying according to light chain (LC) sequence and structure, either leading to crystal formation and Fanconi syndrome or inducing partial inhibition of proximal tubule function. PMID- 12552517 TI - Early referral in chronic kidney disease: an enormous opportunity for prevention. PMID- 12552518 TI - A 66-year-old woman with progressive renal insufficiency, nephrotic syndrome, and monoclonal protein. PMID- 12552520 TI - Ace inhibitors and survival in dialysis patients: effects on serum potassium? PMID- 12552521 TI - Is the evidence for high hematocrit targets valid? PMID- 12552523 TI - Levofloxacin-induced granulomatous interstitial nephritis. AB - The authors report the case of a 47-year-old woman in whom a systemic illness developed characterized by fever, malaise, abnormal liver function results, and acute renal failure after treatment for presumed urinary tract infection with levofloxacin. Because of suspicion of an allergic drug reaction, all medications were discontinued, but the patient remained febrile with renal failure for 18 days. Complete workup for presumed vasculitis, autoimmune illness, or infectious etiologies was negative, and the patient underwent both renal and liver biopsy. Liver biopsy results showed nonspecific changes. Renal biopsy disclosed extensive granulomatous interstitial nephritis with associated granulomatous vasculitis. The patient was begun on oral steroids with rapid defervescence of fever and progressive normalization of renal function. The authors discuss the association of granulomatous nephritis with drugs and review the known nephrotoxicity of fluoroquinolones. PMID- 12552526 TI - Activation and functionalization of molecular nitrogen by metal complexes. AB - Molecular nitrogen is intrinsically unreactive, so much so that it has confounded chemists for decades in attempts to functionalize this abundant diatomic molecule. While biological systems and industrial processes can fix nitrogen to form ammonia, the challenge is to discover a process that involves a homogeneous catalyst that can utilize N(2) as a feedstock to generate higher value organonitrogen materials. In this review, the activation of molecular nitrogen by transition metal complexes is reviewed with the view to present new kinds of transformations for coordinated dinitrogen. Moreover, some reaction types that are as yet unknown are outlined to try and stimulate further research in this area. PMID- 12552527 TI - Indanoyl amino acid conjugates: tunable elicitors of plant secondary metabolism. AB - The unassuming nature of plants belies their viciously effective defensive strategies in the face of herbivore attack. Under the direction of, among others, octadecanoid hormones, plants respond by producing phytoalexins, bitter and toxic alkaloids, protease inhibitors, and even volatile compounds that call predatory insects to the herbivores. A rational design of 4-oxoindanoyl amino acid conjugates based on the phytotoxin, coronatine, as a structural guide resulted in a series of highly active compounds which turn on defensive systems in much the same way as octadecanoid hormones. The developments in the syntheses of indanoyl amino acid conjugates have created easy access to substantial amounts of a variety of such compounds. When these compounds were tested in biological systems, they showed abilities to induce defensive responses that surpassed octadecanoid hormones. In addition, small changes in the structures of these compounds resulted in large differences in the particular defensive systems that were activated. Indanoyl amino acid conjugates are promising tools in photoaffinity approaches towards the macromolecular targets of octadecanoids and their subcellular localization. Owing to the strong activation of plant defense or their efficient induction of fruit abscission which facilitates mechanical harvest, the compounds are promising candidates for future application in agriculture. PMID- 12552528 TI - Seeing what was unseen: new analytical methods for molecular imaging. AB - For nondestructive analysis of chemical processes in living cells, we developed novel intracellular fluorescent indicators for second messengers, protein phosphorylation, and protein/protein interactions that work in single living cells. Key molecules and steps of cellular signaling pathways were visualized under a confocal laser microscope in target live cells using developed fluorescent indicators. A second new approach to molecular imaging is also described. When chemically modified tips were used for STM measurements, contrast enhancements at specific regions in the STM images occurred on the basis of hydrogen bond and metal-coordination interactions. This enabled us to detect not only the distribution of specific chemical species and functional groups but also the orientation of functional groups. The contrast enhancements reflect the increase in a tunneling current due to the overlap of electronic wave functions induced by the chemical interactions between tip and sample. PMID- 12552529 TI - Alkyl chains acting as entropy reservoir in liquid crystalline materials. AB - The roles played by the conformational disordering of alkyl chains in determining the aggregation states of matter are reviewed for liquid crystalline materials from a thermodynamic perspective. Entropy, which is one of the most macroscopic concepts but which has a clear microscopic meaning, provides crucial microscopic information for complex systems for which a microscopic description is hard to establish. Starting from structural implication by absolute (third-law) entropy for crystalline solids, the existence of successive phase transitions caused by the successive conformational melting of alkyl chains in discotic mesogens is explained. An experimental basis is given for the "quasi-binary picture" of thermotropic liquid crystals, i.e., the highly disordered alkyl chains behave like a second component (solvent). A novel entropy transfer between the "components" of a molecule and the resulting "alkyl chains as entropy reservoir" mechanism are explained for cubic mesogens. PMID- 12552530 TI - Substrate effect on the formation of hydrogels with heterogeneous network structure. AB - It was found that when an aqueous solution of vinyl monomers is polymerized on a hydrophobic substrate, obvious heterogeneity occurs in the region of the interface. This substrate effect was observed on polytetrafluroethylene (Teflon), polypropylene (PP), polyethylene (PE), polystyrene (PS), and polyvinylchloride (PVC), but not on hydrophilic substrates. Compared with synthesis on hydrophilic surfaces, the surfaces of hydrogels synthesized on a hydrophobic substrate exhibit a larger degree of swelling, a lower surface coefficient of friction and elastic modulus, weaker interfacial adhesion, and reduced interaction with biological cells. This substrate effect has been observed for many types of aqueous monomer solutions. It was found that the above properties are related to the loosely cross-linked architecture, containing some graft-like polymer chains, that is formed on the gel surface when the gel is prepared on a hydrophobic substrate. To understand the mechanism of the substrate effect, two novel optical methods, electric speckle pattern interferometry (ESPI) and real-time laser sheet refraction (RT-LSR), were developed. It was found that oxygen trapped in the composite interface between the monomer solution and rough hydrophobic substrates played an important role in the substrate effect. PMID- 12552531 TI - Functional tuning of nucleic acids by chemical modifications: tailored oligonucleotides as drugs, devices, and diagnostics. AB - Chemical modifications of nucleic acids present vast opportunities for extending the functions and properties of these biomolecules. In general, efforts invested in this direction pertain to the introduction of reactive functional groups for further derivatizations of oligonucleotides with numerous reporter groups and for equipping nucleic acids with catalytic chemical moieties. This review deals with representative chemical modifications in the nucleobases, sugars, and the phosphate ester backbone and their application from novel catalytic RNA selection to nucleic acid-based biosensors. PMID- 12552532 TI - Nonaqueous magnesium electrochemistry and its application in secondary batteries. AB - A revolution in modern electronics has led to the miniaturization and evolution of many portable devices, such as cellular telephones and laptop computers, since the 1980s. This has led to an increasing demand for new and compatible energy storage technologies. Furthermore, a growing awareness of pollution issues has provided a strong impetus for the science and technology community to develop alternatives with ever-higher energy densities, with the ultimate goal of being able to propel electric vehicles. Magnesium's thermodynamic properties make this metal a natural candidate for utilization as an anode in high-energy-density, rechargeable battery systems. We report herein on the results of extensive studies on magnesium anodes and magnesium insertion electrodes in nonaqueous electrolyte solutions. Novel, rechargeable nonaqueous magnesium battery systems were developed based on the research. This work had two major challenges: one was to develop electrolyte solutions with especially high anodic stability in which magnesium anodes can function at a high level of cycling efficiency; the other was to develop a cathode that can reversibly intercalate Mg ions in these electrolyte systems. The new magnesium batteries consist of Mg metal anodes, an electrolyte with a general structure of Mg(AlX(3-n)R(n)R')(2) (R',R = alkyl groups, X = halide) in ethereal solutions (e.g., tetrahydrofuran, polyethers of the "glyme" family), and Chevrel phases of MgMo(3)S(4) stoichiometry as highly reversible cathodes. With their practical energy density expected to be >60 Wh/Kg, the battery systems can be cycled thousands of times with almost no capacity fading. The batteries are an environmentally friendly alternative to lead-acid and nickel-cadmium batteries and are composed of abundant, inexpensive, and nonpoisonous materials. The batteries are expected to provide superior results in large devices that require high-energy density, high cycle life, a high degree of safety, and low-cost components. Further developments in this field are in active progress. PMID- 12552534 TI - Simultaneous bilateral carotid stenting for postendarterectomy restenosis. AB - We report the successful treatment of bilateral postendarterectomy restenosis using simultaneously placed, bilateral carotid SMART stents and balloon angioplasty. Technical aspects and the results of 29-month follow-up are presented. The benefits derived from single-setting bilateral carotid stenting versus staged bilateral stenting are discussed. PMID- 12552533 TI - Initial multicenter experience with a novel distal protection filter during carotid artery stent implantation. AB - Atheroembolization resulting in transient or permanent neurologic impairment is the most common complication of catheter-based percutaneous carotid artery intervention. Protection of the distal cerebral vasculature during carotid stent implantation may enhance procedural safety. Carotid stent implantation with distal cerebral protection using the FilterWire EX was performed in 35 consecutive patients undergoing 36 procedures at six centers. The FilterWire was delivered and deployed successfully in all 36 cases, and embolic material was retrieved from 74% of procedures. The 30-day rate of major adverse events (death, major or minor stroke) was 0%. Transient ipsilateral periprocedural neurologic ischemia developed in two patients (5.7%), both resolving within 30 min. Distal cerebral protection with the FilterWire during carotid stenting is feasible and safe, results in capture and extraction of atheroembolic debris in the majority of patients while affording uninterrupted cerebral perfusion, and in this initial multicenter experience was associated with a high rate of procedural success without major complications. PMID- 12552535 TI - Brachytherapy for renal artery in-stent restenosis. AB - We describe a 66-year-old female who presented with recurrent acute pulmonary edema and uncontrolled hypertension. She was diagnosed with left renal artery stenosis and treated with angioplasty and stent placement. Her clinical status improved initially but symptoms recurred within 4 months. Further evaluation documented renal artery in-stent restenosis, which was successfully treated with cutting balloon angioplasty followed by brachytherapy. The feasibility of renal artery brachytherapy and short-term follow-up is presented. PMID- 12552536 TI - Directional coronary atherectomy vs. rotational atherectomy for the treatment of in-stent restenosis of native coronary arteries. AB - Management of in-stent restenosis has become a significant challenge in interventional cardiology. Since the mechanism of in-stent restenosis is predominantly intimal hyperplasia, debulking techniques have been used to treat this condition. This study is a nonrandomized comparison of the immediate and long-term results of directional coronary atherectomy (DCA; n = 58) vs. high speed rotational atherectomy (ROTA; n = 61) for the treatment of in-stent restenosis of native coronary arteries. There were no in-hospital deaths, Q-wave myocardial infarctions, or emergency coronary artery bypass surgery in either group. DCA resulted in a larger postprocedural minimal luminal diameter of (2.57 +/- 0.51 vs. 2.14 +/- 0.37 mm; P < 0.0001) and a larger acute gain (1.83 +/- 0.52 vs. 1.42 +/- 0.48 mm; P < 0.0001). Furthermore, 12-month clinically indicated target lesion revascularization (39% vs. 21%; P = 0.02) and long-term follow-up MACE (44% vs. 28%; P = 0.03) was greater in the ROTA group. The present study suggests that DCA appears to be superior to ROTA for the treatment of in-stent restenosis of native coronary arteries. Compared to ROTA, the debulking effect of DCA leads to a larger postprocedure minimal luminal diameter, and a lower incidence of subsequent target lesion revascularization and MACE. PMID- 12552537 TI - Relationship between neointimal regrowth and mechanism of acute lumen gain during the treatment of in-stent restenosis with or without supplementary intravascular radiation. AB - We investigated whether neointimal regrowth is related to the mechanism of acute lumen gain during the treatment of in-stent restenosis (ISR) lesions both with and without adjunct intravascular brachytherapy. From the WRIST (Washington Radiation for In-Stent Restenosis Trial) cohort, 54 ISR patients ((192)Ir, 29; placebo, 25) were treated with nonrepeat stenting percutaneous interventions (excimer laser, rotational atherectomy, and/or balloon angioplasty) prior to (192)Ir or placebo therapy. Using Simpson's method, serial volumetric intravascular ultrasound (IVUS) analyses (pre- and posttreatment and 6-month follow-up) were analyzed to obtain stent, lumen, and intimal hyperplasia (IH) volumes that were then adjusted for stent length to create stent, lumen, and IH volume indexes. In the placebo group, the acute reduction of neointima (1.6 +/- 1.4 mm(3)/mm) was counteracted by intimal regrowth (2.1 +/- 1.7 mm(3)/mm). The amount of intimal regrowth correlated directly with the intimal reduction due to the intervention (r = 0.76; P < 0.001), but not with the amount of additional stent expansion. In the (192)Ir-treated group, intimal regrowth was significantly less than in the placebo group (-0.3 +/- 0.1 vs. 2.1 +/- 1.7 mm(3)/mm; P < 0.001) despite a similar initial intimal reduction (1.3 +/- 0.9 vs. 1.6 +/- 1.4 mm(3)/mm; P = NS). No correlation was found between intimal reduction at the time of the procedure and intimal regrowth in the (192)Ir group. In this study, neointimal regrowth following treatment of ISR lesions correlates directly with the extent of acute intimal volume reduction, but not with the extent of additional stent expansion. This relation is not seen in ISR segments treated with radiation, where intimal regrowth is substantially inhibited. PMID- 12552538 TI - Lesion length assessment using a new vessel caliper. AB - Assessment of coronary lesion length using visual estimation or quantitative coronary angiography can be unreliable. We tested the accuracy of a new handheld caliper device to measure lesion length in a bench-top model and in eight patients undergoing percutaneous coronary intervention (PCI). Caliper-derived length measurements were compared to the known reference distance in the bench top model and visual or intravascular ultrasound (IVUS)-derived measurements in vivo. In the coronary model, caliper-derived measurements were accurate and correlated well with known reference distances regardless of the angiographic projection. During PCI, there was a poor correlation between the best visual estimate of length and IVUS-derived measurements. In contrast, caliper-derived measurements correlated closely with IVUS-derived measurements. This handheld caliper provides a simple and accurate method of assessing intracoronary lesion length and may be particularly useful during coronary stenting and when adjunctive brachytherapy is performed. PMID- 12552539 TI - Impact of intracoronary radiation on in-stent restenosis involving ostial lesions. AB - The aim of this study was to compare 6-month clinical outcomes of patients with in-stent restenosis (ISR) involving the ostium treated with intracoronary radiation therapy (IRT) compared to placebo therapy, and also to nonostial lesions treated with IRT. Coronary interventions in ostial lesions have a high rate of recurrence of restenosis. The impact of IRT on ostial ISR has been inadequately characterized. We assessed patients enrolled in gamma (192-iridium) and beta (90-yttrium, 32-phosphorus) radiation trials for ISR at the Washington Hospital Center. Of patients receiving IRT, 105 (8%) patients had ostial ISR and 1,289 (92%) patients had nonostial ISR. Twenty-seven patients had ostial ISR and received placebo therapy. Baseline demographic and angiographic and procedural details were similar, except ostial IRT patients had a trend toward shorter lesions (15.4 +/- 10.8 vs. 24.1 +/- 12.2 mm; P < 0.001) and had a higher rate of saphenous vein graft disease (46% vs. 19%; P < 0.001) compared to nonostial IRT patients. At 6 months, ostial lesions treated with IRT for ISR had a reduced rate of target lesion revascularization (TLR) compared to ostial lesions treated with placebo (15% vs. 43%; P = 0.004). Outcomes at 6 months were similar for the ostial and nonostial IRT groups including TLR (15% vs. 14%; P = 0.80) and composite major adverse cardiac events (18% vs. 15%; P = 0.46). Intracoronary radiation therapy is effective for ostial in-stent restenotic lesions and should be comfortably used for this challenging anatomic location. PMID- 12552540 TI - Restick following initial Angioseal use. AB - The Angioseal hemostatic device is currently in widespread use for arterial closure after both diagnostic and interventional procedures. Resticking of the artery in which an Angioseal device has been placed has been discouraged for up to 90 days after the initial device placement because of theoretical concerns of disruption or dislodgment of the hemostatic plug. However, no data are available to address this concern. We evaluated the incidence of vascular complications following restick of the artery in which an Angioseal device had been deployed C in TSC1and 1462-28del42bp, 1774del4bp, and N1643K (4947C>G) in TSC2, which were present in only 6.5-17% of the patients' alleles. We have also applied this technique to successfully resolve seven somatic APC mutations in colorectal tumor samples that were previously undetectable by direct PCR product sequencing. This method may simplify many of the currently challenging goals in mutation detection. PMID- 12552558 TI - High-throughput single strand conformation polymorphism mutation detection by automated capillary array electrophoresis: validation of the method. AB - Capillary array electrophoresis (CAE) is a novel technique, which allows for high throughput analysis of DNA fragments. When screening for mutations in whole populations or large patient groups it is necessary to have robust and well characterized setups for high throughput analysis. For large-scale mutation screening, we have developed procedures for single strand conformation polymorphism (SSCP) assays using CAE (CAE-SSCP) whereby we may increase both the sensitivity and the throughput compared to conventional SSCP analysis. In this study we have validated CAE-SSCP by 1) comparing detection by slab-gel based SSCP with CAE-SSCP of mutations in the MYH7, MYL2, and MYL3 genes encoding sarcomere proteins from patients suffering from hypertrophic cardiomyopathy; and 2) by constructing a series of 185 mutants having substitution mutations, as well as insertion/deletion mutations, or some combinations of these, in different sequence contexts in four exons and different positions relative to the end of the amplicon (three from the KCNQ1 gene, encoding a cardiac potassium channel, and one from the TNNI3 gene encoding cardiac troponin I). The method identified 181 out of 185 mutations (98%), and the data suggest that the position of mutation in the fragment had no effect on the sensitivity. Analysis of the specificity of the method showed that only very few mutants could not be distinguished from each other and there were no false positives. PMID- 12552559 TI - Comprehensive scanning of the ATM gene with DOVAM-S. AB - Mutation detection at the ATM locus has been difficult because of the large size of the gene (66 exons), the fact that mutations are located throughout the entire gene with no hotspots, and the difficulty of distinguishing mutations from polymorphisms. In this study, the entire coding region (exons 4-65) was scanned, as well as the adjacent intronic regions, using DOVAM-S (Detection Of Virtually All Mutations-SSCP), a robotically-enhanced, multiplexed scanning method that is a highly sensitive modification of SSCP. Forty-three unrelated patients and four obligate carriers were studied. Of the 90 expected mutant alleles, 71 were identified (79%). The mutations included 17 nonsense (24%), 20 frameshift (28%), 20 splice (28%), 10 missense (14%), one in-frame deletion (1%), and three that alter the initiation codon (4%). Among the ataxia-telangiectasia patients, two potentially causative mutations were identified in 30 individuals: 22 had two truncating mutations, four had one truncating and one missense mutation, three had two missense mutations, and one had a truncating mutation and an in-frame deletion of three amino acids. For seven A-T patients and all four obligate carriers, only one truncating mutation was detected. Six of the 43 A-T patients had no detected mutations (14%). Twelve novel mutations and six novel polymorphisms were detected. The results of this complete scan of the ATM coding region showed that 86% of causative ATM mutations were truncating and 14% were missense. DOVAM-S is a rapid, efficient method of performing A-T diagnosis and carrier testing on a clinical time scale. PMID- 12552560 TI - Rapid detection of exon 1 NRAS gene mutations using universal heteroduplex generator technology. AB - Specific NRAS oncogene missense mutations have been frequently found in some tumors and several hematological diseases, especially in those of myeloid origin. There is a wide range of PCR-based methods for screening and detection of NRAS exon 1 single-base substitutions. However, there are disadvantages and ambiguities associated with these techniques because all of them require either separate probes, separate PCR amplifications, or complicated post-PCR manipulations. This report describes a new approach for detection of NRAS gene mutations at codon 12 and 13 based on the DNA heteroduplex analysis method. The strategy relies upon differential electrophoretic behavior of induced heteroduplex molecules formed by cross-hybridization of two PCR-amplified species, the sample under analysis and the synthetic universal heteroduplex generator (UHG). The screening of a panel of all codon 12 and 13 NRAS mutant DNA variants indicated that this approach discriminates all 12 relevant mutations. The sensitivity of the method was estimated by a competitive assay where mutant alleles could be detected at a dilution level of 1 to 16 wild-type alleles. This UHG technology was tested on some clinical samples previously studied by PCR-ASO. This methodology is highly specific, sensitive, and achieves an appreciable reduction in workload and time because it requires one PCR amplification followed by polyacrylamide gel electrophoresis in standard conditions. We propose that this new approach may be applied as an alternative strategy for codon 12-13 NRAS mutations and it could be easily incorporated into the range of routine assays performed in oncology laboratories. PMID- 12552561 TI - Missense mutations in the DNA-binding region and termination codon in PAX6. AB - We have identified nine novel intragenic mutations of the PAX6 gene in 30 patients with aniridia. One patient with Wilms' tumor, aniridia, genitourinary anomalies, and mental retardation (WAGR syndrome) had deletion of 11p and had lost the paternal PAX6 allele. Two patients had small deletions: a frameshift that should result in early termination of the PAX6 protein, and a frameshift that leads to a termination-site change and run-on into the 3' untranslated region (UTR). The other 27 patients had single base-pair mutations. Four had splicing defects; three had IVS6+1G>A, which was at a mutation hotspot in the PAX6 gene; 10 had premature termination (four 1024C>T [R203X], also at a mutation hotspot); and six had missense mutations. Missense mutation A321T (1378G>A) was a polymorphic change; the other five missense mutations were L46R, C52R, I56T, G73D, and I87K. These five codons are in the PAX6 paired domain and are highly conserved throughout the entire paired family. Seven patients had a mutation in the normal stop codon (TAA). This change leads to run-on into the 3' UTR and is also at a mutation hotspot. All 30 mutations should result in PAX6 haploinsufficiency. No correlation was observed between mutation sites and phenotypes. PMID- 12552562 TI - The Iranian Human Mutation Gene Bank: a data and sample resource for worldwide collaborative genetics research. AB - As Human Genome Project exploration continues, the necessity of having a broader spectrum of genomic DNA material from different nationalities to study various aspects of hereditary disease becomes more obvious. The existence of high genetic polymorphism within and between different communities in the world makes it necessary for the gene hunters to investigate many different populations. Iran, a large country with close to 66 million people, is a land of different nationalities, tribes, and religions that offers a highly heterogeneous gene pool to the genetics researcher. The purity of many different races in this country has been highly conserved by geographical borders and by an ancient culture that has always encouraged intrafamilial marriages. All these have created a population that is remarkably heterogeneous yet high in consanguinity rate. During the last five years of investigation we have established a DNA bank, the Iranian Human Mutation Gene Bank (www.IHMGB.com), which contains all genetic diseases studied in Iran that have the Mendelian mode of inheritance. Some of the samples are assigned to common or novel mutations and others belong to patients with clinical profiles associated with particular genetic diseases but undefined mutation. This bank stores samples of DNA from the patient and his/her first degree relatives together with a comprehensive pedigree and clinical profile for each sample. To facilitate collaboration with other scientists around the world with the same interests, we decided to present our experimental projects online. This DNA bank provides opportunities for us to collaborate with scientists outside Iran. It offers a sample resource to research scientists around the world, at no charge, for the purpose of investigating the various aspects of genetic disorders from prenatal diagnosis to gene structure and function. It is strongly stressed that no commercial benefit is involved in the establishment of this DNA bank and the DNA samples are free of charge. However, to meet our goals and to respect ethical values, DNA samples can only be used under certain conditions stated in the User Consent Form. PMID- 12552563 TI - Novel mutations in the TCIRG1 gene encoding the a3 subunit of the vacuolar proton pump in patients affected by infantile malignant osteopetrosis. AB - Fifty percent of the infantile malignant osteopetrosis (IMO) cases reported in the literature present mutations in the TCIRG1 gene encoding the 116-kDa osteoclast specific subunit of the vacuolar proton ATPase (ATP6I). In this study, we identified four novel mutations in a series of six IMO patients. All of these mutations correspond to single nucleotide changes and affect splice acceptor or donor sites, resulting in aberrant transcription products. We report also a missense mutation, G405R, previously described in several Costa Rican patients. This independent finding suggests that the highly conserved residue at amino acid 405 plays a critical role in the a3 subunit function. Finally, the results of this study were used to provide a prenatal diagnosis to one of the families. PMID- 12552564 TI - Spectrum of sequence variation in the FANCG gene: an International Fanconi Anemia Registry (IFAR) study. AB - Fanconi anemia (FA) is a genetically heterogeneous autosomal recessive syndrome associated with chromosomal instability, hypersensitivity to DNA cross-linking agents, and predisposition to malignancy. The gene for FA complementation group G (FANCG) was the third FA gene to be cloned, and was found to be identical with human XRCC9, which maps to 9p13. The cDNA is predicted to encode a polypeptide of 622 amino acids, with no sequence similarities to any other known protein or motifs that could point to a molecular function for FANCG/XRCC9. We used single strand conformational polymorphism analysis (SSCP) to screen genomic DNA from a panel of 307 racially and ethnically diverse unrelated FA patients from the International Fanconi Anemia Registry (IFAR) for variants in FANCG. Twenty-seven abnormal SSCP patterns were found; 18 of these variants appear to be pathogenic mutations while nine are likely to be nonpathogenic polymorphisms. Direct sequencing of genomic DNA from seven FA-G probands with one mutant allele not detected in the SSCP study and three additional probands assigned to the FA-G complementation group by retroviral correction with FANCG resulted in the detection of nine additional pathogenic mutations and two common SNPs. Conditions for rapid screening for these mutations by DHPLC for use in a clinical laboratory setting were established. The most common FANCG mutations in the IFAR population were: IVS8-2A>G (seven Portuguese-Brazilian probands), IVS11+1G>C (seven French Acadian probands), 1794_1803del10 (seven European probands), and IVS3+1G>C (five Korean or Japanese probands). Our data suggest that the Portuguese-Brazilian, French-Acadian, and Korean/Japanese mutations were likely to have been present in a founding member of each of these populations. PMID- 12552565 TI - Mutations in the CACNA1F and NYX genes in British CSNBX families. AB - X-linked congenital stationary night blindness (CSNBX) is a genetically and phenotypically heterogeneous non-progressive disorder, characterised by impaired night vision but grossly normal retinal appearance. Other more variable features include reduction in visual acuity, myopia, nystagmus and strabismus. Genetic mapping studies by other groups, and our own studies of British patients, identified key recombination events indicating the presence of at least 2 disease genes on Xp11. Two causative genes (CACNA1F and NYX) for CSNBX have now been identified through positional cloning strategies. In this report, we present the results of comprehensive mutation screening in 14 CSNBX families, three with mutations in the CACNA1F gene and 10 with mutations in the NYX gene. In one family we failed to identify the mutation after testing RP2, RPGR, NYX and CACNA1F. NYX gene mutations are a more frequent cause of CSNBX, although there is evidence for founder mutations. Our report of patient population mutation screening for both CSNBX genes, and our exclusion of RP2 and RGPR, indicates that mutations in CACNA1F and NYX are likely to account for all CSNBX. PMID- 12552566 TI - Phenotypic cellular characterization of an ataxia telangiectasia patient carrying a causal homozygous missense mutation. AB - Most disease-causing mutations in Ataxia telangiectasia (AT) patients correspond to truncating mutations in the ATM gene with very few cases of AT patients carrying two missense sequence alterations being reported. The cellular phenotype of a lymphoblastoid cell line established from an AT patient (AT173) who showed classical clinical AT features, and carried two homozygous missense alterations, the 378T>A variant and 9022C>T located within the ATM kinase domain, has been characterized. ATM mRNA was detectable and the ATM protein level was approximately 50% of that seen in normal cell lines. Functional analysis of this protein revealed a total absence of ATM kinase activity measured either in vitro or in vivo, before and after exposure to ionizing radiation. The AT173 cell line was hypersensitive to ionizing radiation and exhibited a G1 cell cycle arrest defect and an accumulation of cells in G2 phase of the cell cycle after irradiation, a response that is identical to that seen in AT cell lines carrying truncating mutations. These phenotypic features strongly suggest that the 9022C>T (R3008C) missense mutation is the disease-causing mutation and that the presence of ATM protein is not always predictive of a normal cellular phenotype. PMID- 12552567 TI - Identification of GUCY2D gene mutations in CORD5 families and evidence of incomplete penetrance. AB - Cone rod dystrophy 5 (CORD5) is an autosomal dominant retinal disease that primarily affects cone function. The locus has previously been mapped to human chromosome 17p12-p13 between the markers D17S926/D17S849 and D17S945/D17S804. One of our "unaffected" recombinant individual from family 1175 was subsequently found to cross through this interval. Reexamination revealed that he was in fact mildly affected. This expanded the minimum candidate region. Direct sequencing of the GUCY2D and other candidate genes within this interval was carried out on 2 American families affected with CORD5. There was an R838C missense mutation within the GUCY2D gene in one and a R838H missense mutation in another families. The previously reported mutations for CORD6 are clustered at the same position within the gene. These results indicate that both CORD5 (MIM# 600977) and CORD6 (MIM# 601777) are actually the same disease. We conclude that significant variability in expression and incomplete penetrance exists even within one family. PMID- 12552568 TI - Screening of patients with hereditary spastic paraplegia reveals seven novel mutations in the SPG4 (Spastin) gene. AB - Hereditary spastic paraplegia (HSP) is a heterogeneous condition characterised in its pure form by progressive lower limb spasticity. Mutations in SPG4 (encoding spastin) may be responsible for up to 40% of autosomal dominant (AD) cases. A cohort of 41 mostly pure HSP patients from Britain and Austria, 30 of whom displayed AD inheritance, was screened for mutations in SPG4 by single strand conformation polymorphism (SSCP) analysis followed by sequencing of samples with mobility shifts. We identified eight SPG4 mutations in pure AD HSP patients, seven of which were novel: one missense mutation within the AAA cassette (1633G>T), two splice site mutations (1130-1G>T, 1853+2T>A) and four frameshift mutations (190_208dup19, 1259_1260delGT, 1702_1705delGAAG, 1845delG). A novel duplication in intron 11 (1538+42_45dupTATA) was also detected. We report the HUGO-approved nomenclature of these mutations as well. Furthermore, we detected a silent change (1004G>A; P293P), previously reported as a mutation, which was also present in controls. The frequency of SPG4 mutations detected in pure AD HSP was 33.3%, suggesting that screening of such patients for SPG4 mutations is worthwhile. Most patients will have unique mutations. Screening of SPG4 in apparently isolated cases of HSP may be of less value. PMID- 12552569 TI - NF1 gene analysis based on DHPLC. AB - The high mutation rate at the NF1 locus results in a wide range of molecular abnormalities. The majority of these mutations are private and rare, generating elevated allelic diversity with a restricted number of recurrent mutations. In this study, we have assessed the efficacy of denaturing high-performance liquid chromatography (DHPLC), for detecting mutation in the NF1 gene. DHPLC is a fast and highly sensitive technique based on the detection of heteroduplexes in PCR products by ion pair reverse-phase HPLC under partially denaturing conditions. We established theoretical conditions for DHPLC analysis of all coding exons and splice junctions of the NF1 gene using the WAVEmaker software version 4.1.40 and screened for mutations a panel of 40 unrelated NF1 patients (25 sporadic and 15 familial), genetically uncharacterized. Disruptive mutations were identified in 29 individuals with an overall mutation detection rate of 72.5%. The mutations included eight deletions (exons 4b, 7, 10a, 14, 26, and 31), one insertion (exon 8), nine nonsense mutation (exons 10a, 13, 23.1, 27a, 29, 31, and 36), six missense mutations (exons 15, 16, 17, 24, and 31), four splice errors (exons 11, 14, 36, and 40) and a complex rearrangement within exon 16. Eighteen (62%) of the identified disruptive mutations are novel. Seven unclassified and three previously reported polymorphisms were also detected. None of the missense mutations identified in this study were found after screening of 150 controls. Our results suggest that DHPLC provides an accurate method for the rapid identification of NF1 mutations. PMID- 12552570 TI - BRCA2 germline mutations in Cypriot patients with familial breast/ovarian cancer. AB - Germline mutations in the BRCA2 gene have been shown to be associated with familial female and male breast cancer. Mutations occur throughout the entire coding region of the gene, and there is considerable ethnic and geographical diversity in the deleterious mutations detected in different populations. No data exist on the role of the BRCA2 gene in the Cypriot population. In this study we present the results of characterizing mutations in the BRCA2 gene, in 26 Cypriot families with multiple cases of breast/ovarian cancer. The entire coding region, including splice sites, of BRCA2 were sequenced using cycle sequencing. In total 29 BRCA2 variants were detected which include 3 truncating mutations, 8 missense mutations, 6 polymorphisms and 12 intronic variants. The 3 truncating mutations are frameshift mutation 8984delG (exon 22), and two nonsense mutations, namely C1913X (exon 11) which is a novel mutation, and K3326X (exon 27). It is of interest that frameshift mutation 8984delG was the most frequent, since it was detected in 5 patients from three different families. Among the 6 polymorphisms detected, polymorphism T77T is novel and similarly 4 of the 12 intronic variants were also novel, namely IVS1+8G>A, IVS1-96insA, IVS4+36A>G and IVS11-51G>T. These results show that deleterious BRCA2 mutations, occur at the same frequency, about 20%, in Cypriot families, as that recorded in other European populations. We conclude that the BRCA2 gene plays a significant role in the familial breast cancer phenotype in the Cypriot population. PMID- 12552571 TI - Functional analysis of LKB1/STK11 mutants and two aberrant isoforms found in Peutz-Jeghers Syndrome patients. AB - Peutz-Jeghers Syndrome (PJS) is thought to be caused by mutations occurring in the widely expressed serine/threonine protein kinase named LKB1/STK11. Recent work has led to the identification of four mutants (R304W, I177N, K175-D176del, L263fsX286) and two novel aberrant LKB1/STK11 cDNA isoforms (r291-464del, r485 1283del) in a group of PJS Italian patients. Three of the four mutations only change 1 or 2 amino acids in the LKB1/STK11 catalytic domain. Here we demonstrate that all six LKB1/STK11 variants analysed are completely inactive in vitro as they were unable to autophosphorylate at Thr336, the major LKB1/STK11 autophosphorylation site, and to phosphorylate the p53 tumour suppressor protein. We also show that 5 out of the 6 variants are entirely localised in the nucleus in contrast to the wild type LKB1/STK11, which is detected in both the nucleus and cytoplasm. Finally we demonstrate that all 6 LKB1/STK11 variants, in contrast to wild type LKB1/STK11, are unable to suppress the growth of melanoma G361 cells. Taken together, these results demonstrate that the LKB1 mutations investigated in this study lead to the loss of serine/threonine kinase activity and are therefore likely to be the primary cause of PJS development in the patients that they were isolated from. PMID- 12552572 TI - Effect of husbands' education on fatness of wives. AB - The aim of the study was to examine the relationship between social position of females achieved by marriage and level of fatness and relative fat distribution. The data of 588 healthy, occupationally active, married women, age 21-62 years, with 12 years of education (completed secondary school) were used. The body mass index (BMI, kg/m(2) ), triceps and subscapular skinfold thicknesses, and summed skinfold thicknesses were used as indicators of fatness. The waist-hip ratio, the waist-thigh ratio, and waist, hip, and thigh circumferences were used as indicators of fat distribution. According to the educational level of husbands, women were grouped as 1). moving up the social scale (spouse with complete university education), 2). stable (equal level of education), and 3). moving down the social scale (spouse who never passed beyond the level of basic vocational school, i.e., skilled and unskilled manual workers). The two opposite groups were analyzed, i.e., moving up and moving down. Women with secondary schooling who married up were consistently leaner than women who married down. A similar pattern was observed for fat distribution. Women marrying down had more abdominal body fat compared to women marrying up. PMID- 12552573 TI - Potential association between endogenous leptin and sympatho-vagal activities in young obese Japanese women. AB - Leptin is an adipocyte-derived hormone that decreases food intake and increases energy expenditure through the activation of the sympathetic nervous system (SNS). Notwithstanding recent intensive research, the underlying physiological mechanism of leptin as well as the etiology of obesity in humans remains elusive. The present study attempted to investigate the potential association between endogenous circulating leptin and sympatho-vagal activities in age- and height matched obese and nonobese healthy young women. Plasma leptin concentrations were measured by radioimmunoassay. The autonomic nervous system activity was assessed during the resting condition by means of a recently devised power spectral analysis of heart rate variability, which serves to identify three separate frequency components, very low (VLO), low (LO), and high (HI). Plasma leptin concentrations were greater in the obese than in the control group (45.7 +/- 5.89 vs. 11.2 +/- 1.10 ng. ml(-1), P < 0.01). As to the contribution of endogenous leptin to SNS activity, both the ratios of the VLO frequency component reflecting thermoregulatory sympathetic function and the global SNS index [(VLO + LO)/HI] to plasma leptin concentration were markedly reduced in the obese compared to the control group (VLO per leptin: 5.9 +/- 1.39 vs. 37.8 +/- 8.1 ms(2). ml. ng(-1), P < 0.01; SNS index per leptin: 0.04 +/- 0.008 vs. 0.33 +/- 0.01 ml c. ng(-1), P < 0.01). Additionally, a nonlinear regression analysis revealed that these ratios exponentially decreased as a function of body fat content (VLO per leptin r(2) = 0.57, P < 0.01; SNS index per leptin r(2) = 0.53, P < 0.01). Our data suggest that reduced sympathetic responsiveness to endogenous leptin production, implying peripheral leptin resistance, might be a pathophysiological feature of obesity in otherwise healthy young women. The findings regarding the association of leptin, body fat content, and SNS activity further indicate that the 30% of total body fat, which has been used as a criterion of obesity, might be a critical point at which leptin resistance is induced. PMID- 12552574 TI - Heritability and anthropometric influences on human fertility. AB - This study researched the impact of anthropometrics and size-of-family of orientation on women's fertility by using path analysis. The data were collected as part of the anthropological study conducted in Ireland by Harvard University personnel before the Second World War. The women included in this analysis were all over age 49 and were either married or widowed at the time of the survey. Our results indicate that the heritability of fertility is moderate in this sample and that there is a tendency for heavy women to have a higher fertility. However, when anthropometrics and size-of-family of orientation were entered as independent variables in a path diagram, an insignificant portion of the variation of fertility was explained. In this Irish population, the main cause of differential fertility was cultural rather than biological. A large portion of women never married and no unmarried woman reported producing a child. PMID- 12552575 TI - Genetic structure of seven Mexican indigenous populations based on five polymarker loci. AB - This descriptive study investigates the genetic structure of seven Mexican indigenous populations (Mixteca Alta, Mixteca Baja, Otomies, Purepecha, Nahuas Guerrero, Nahuas-Xochimilco, and Tzeltales) on the basis of five PCR-based polymorphic DNA loci: LDLR, GYPA, HBGG, D7S8, and GC. Genetic distance and diversity analyses indicate that these Mexican indigenous are similar and that more than 96% of the total gene diversity (H(T)) can be attributed to individual variation within populations. Mixteca-Alta, Mixteca-Baja, and Nahuas-Xochimilco show indications of higher admixture with European-derived persons. The demonstration of a relative genetic homogeneity of Mexican Indians for the markers studied suggests that this population is suitable for studying disease marker associations in the search for candidate genes of complex diseases. PMID- 12552576 TI - Beta-globin gene cluster haplotypes as evidence of African gene flow to the northeastern coast of Venezuela. AB - In order to study the origin of mutation HBB*S in Sucre and Anzoategui states and the genetic affinities of these Venezuelan populations with other human groups, the beta-globin gene cluster haplotypes were determined for 28 sickle cell and/or S-beta thalassemia patients and for 37 individuals with normal hematological parameters. Bantu, Benin, Senegal, and atypical haplotypes were identified in 50%, 36%, 2%, and 12% of the HBB*S chromosomes, respectively. Similar results have been published for Venezuelan patients from the central states, but a different trend is shown in a publication based on a group of patients from different regions of the country. For HBB*A, haplotype 2 (+ - - - -), characteristic of non-African groups, was the most common (39%), followed by haplotype 3 (- - - - +) of African origin, and haplotype 6 (- + + - +), also typical of non-Africans. The results reveal a high level of admixture of the Sucre-Anzoategui population. The importance of specific conditions which have acted differently in the Venezuelan populations, such as founder effect, genetic drift, isolation, and endogamy are discussed. Genetic distances between the Sucre Anzoategui sample and several other human populations calculated on the basis of the HBB*S and HBB*A haplotypes revealed similar results, the closest genetic relationships being observed in relation to Bantu-speaking groups. These results confirm the utility of the beta-globin haplotypes for population studies and contribute to knowledge of the Venezuelan gene pool. PMID- 12552577 TI - Risk factors for hysterectomy among Mexican-American women in the US southwest. AB - The purpose of this study was to assess risk factors associated with a history of hysterectomy among Mexican-American women living in the United States Southwest. Mexican-American women ages 20-74 at time of interview were defined as a subpopulation among adults in the Hispanic Health and Nutrition Examination Survey (HHANES), 1982-1984. Language preference, reproductive history, level of education, poverty status, generation of immigration, marital status, and insurance coverage were examined in relation to risk of hysterectomy using weighted tabulation and logistic regression for data resulting from complex survey designs. Heretofore, language preference has not been a variable considered in relation to risk of hysterectomy. In the HHANES, over 60% of women who spoke English most often rather than Spanish reported a history of hysterectomy. Women who had previously been pregnant were almost four times as likely (odds ratio 3.972) to have had a hysterectomy compared to women who had never been pregnant. Women who expressed any preference for English were twice as likely (odds ratio 2.050) to have had a hysterectomy than were those who responded that they exclusively preferred Spanish. Age, higher levels of education, and higher economic status also increased the risk of hysterectomy. In contrast, reproductive history, marital status, prior tubal ligation, generation of immigration, and health insurance did not have substantial effects on the risk of hysterectomy. This study suggests that, in the future, the effect of language preference should not be overlooked when considering risk factors for hysterectomy. PMID- 12552578 TI - Allometric scaling of strength in an independently living population age 55-86 years. AB - Most physiological functions vary allometrically with body size; however, few investigators have examined the relationship between strength and body size with allometric scaling. Thus, we hypothesized that allometric analysis would reveal that both the amount and quality of muscle are significant determinants of strength in the elderly. Allometric analyses were used to determine the influence of limb cross-sectional area (CSA), physical activity, demispan (distance between index-middle finger web and the sternal notch), leg length, and sex on grip and plantar flexor strength in men (n = 188) and women (n = 205) age 55-86 years. Physical activity was measured using a self-reporting questionnaire (Taylor et al. [1978] J Chron Dis 31:741-755). Forearm and leg CSA was estimated from anthropometry. There was an age-related decline in grip strength, independent of forearm CSA, demispan, and sex, equal to approximately 12% per decade, whereas plantar flexor strength adjusted for leg CSA, physical activity, and sex was reduced at a rate of approximately 15% per decade. The allometric models explained 71.4% (r = 0.845) and 38.8% (r = 0.623) of the variance in grip and plantar flexor strength, respectively. Model parameters were identified using multiple linear regression (P < 0.05). Thus, grip strength = forearm CSA(0.435). demispan(0.161). exp(3.905 - 0.012 age + 0.413 sex) and plantar flexor strength = leg CSA(0.223). physical activity (0.115). exp(5.867 - 0.015 age + 0.366 sex). These findings indicate that age-related reductions in muscle CSA do not fully account for strength declines with age. Physical activity is also important and partially explains these reductions. PMID- 12552579 TI - Minor physical anomalies in mentally healthy subjects: Internal consistency of the Waldrop Physical Anomaly Scale. AB - The aim of the study was to investigate the prevalence of minor physical anomalies in mentally healthy subjects by using the Waldrop Physical Anomaly Scale and to assess the reliability (internal consistency) of the scale. The subjects were 82 mentally healthy individuals (42 men, 40 women) of Bulgarian origin who were examined for minor physical anomalies. Mentally healthy individuals show a low mean score of minor physical anomalies. The anomalies prevail in the craniofacial region. The correlations between the anomalies are low, which implies poor internal consistency of the scale, probably due to the heterogeneity of the anomalies in terms of location, character, and time of prenatal development and adversity. Providing a base for comparative studies of developmental disorders, the findings infer the necessity of a more reliable scale for examination of informative morphogenetic variants which can distinguish between minor malformations and phenogenetic variants and suggest the period of prenatal adversity. PMID- 12552580 TI - Behavior of loci D1S1656 and D12S391 in a sample from Maracaibo, Venezuela. AB - Two recently reported short tandem repeat polymorphisms characterized by PCR, D1S1656 and D12S391, were investigated in a sample from Maracaibo, an admixed population of Venezuela, in order to evaluate their application in forensic and population genetics studies. The unbiased heterozygosities were 0.9011 and 0.8444 for locus D1S1656 and D12S391, respectively. The joint discrimination power and joint probability of exclusion were 0.99972 and 0.93287. When allele frequencies of locus D1S1656 from Maracaibo were compared with eight other populations, our group clustered with the European or European-derived samples, mainly from Spain. In the comparison of locus D12S391 with 16 populations, Maracaibo clustered with 3 Asian samples. The high heterozygosity and discrimination power make these two loci important candidates to be considered for STR packages for forensic and population genetic purposes. PMID- 12552581 TI - Age differences in growth and physical abilities in trained and untrained girls 10-17 years of age. AB - The purpose of the present study was to assess the age differences in growth characteristics and physical abilities of physically active adolescent girls and to compare them to corresponding data for girls nonactive in sport. The cross sectional study was carried out on 643 Estonian girls, 10-17 years of age, who were regularly training track and field, and 902 nonactive girls. The organized physical activity of the studied girls consisted of lessons in the school curriculum (2 x 45 minutes per week) for both groups and special track and field training for the first group. Height, weight, and the body mass index (BMI) were used to characterize growth status. Physical ability was assessed with the 30-m dash, standing long jump, medicine ball throw (2 kg), standing quintuplet jump, isometric strength of back extensor muscles, and 1-minute ergocycling at the highest possible rate. Girls active in track and field were taller in all the age groups (P < 0.05-0.001) and lighter except at 17 years, when they were heavier (P > 0.05), but the differences at 12 and 13 years were not significant. BMI was also significantly lower in active girls (P < 0.05-0.001) in all age groups, except at 17 (P > 0.05). The actively training girls had higher physical abilities at all ages from 10-17. The annual differences in performance scores were significant (P < 0.05-0.001) up to 15 years except for the standing long jump. Differences in mean scores of most motor abilities were minimal or reduced significantly at 13-14 years in nonactive girls, but were significant in active girls. PMID- 12552582 TI - Obesity correlates with increased blood pressures in urban Native American youth. AB - Although obesity is a growing problem with Native American youth living on reservations, little research has been conducted examining the prevalence of obesity and correlations between age, body composition, dietary intake, and blood pressures (BP) for urban Native youth. The purpose of this study was to investigate the relationship of these variables in urban Native American youth. Height and weight were measured for 155 Native American youth, age 5-18 years, and the body mass index (BMI) was calculated and classified into percentile categories. Skinfold thicknesses at the biceps, triceps, suprailiac, and subscapular sites, arm and waist circumferences, and systolic (SBP) and diastolic (DBP) blood pressures were also measured. There was a high proportion of obesity (>95 percentile) for youth in all age groups. The prevalence was 38% for the 5-10 year-olds and 45% for the 11-18-year-olds youth. There were no significant correlations between SBP and DBP and dietary variables. Mean SBP and DBP increased with increasing BMI percentiles. Stepwise regression analyses showed that waist circumference, age, and BMI were strong predictors for SBP, while waist circumference and age were predictors for DBP in the total sample. The findings suggest that overweight/obesity is very prevalent among urban Native American youth and the increased adiposity is associated with increased SBP and DBP. PMID- 12552583 TI - Physical activity, body fat, and serum C-reactive protein in postmenopausal women with and without hormone replacement. AB - The objective was to determine whether higher physical activity is associated with lower serum C-reactive protein (CRP), independent of oral hormone replacement therapy (HRT) status and body fatness, in 133 postmenopausal women using a cross-sectional exploratory design at a university research laboratory. The subjects were 133 postmenopausal women, age 50-73 years, with no evidence of coronary artery disease or diabetes. The main outcome measures were: serum CRP, physical activity as measured by Stanford 7-day activity recall, body fat (both total and regional) as measured by dual energy X-ray absorptiometry (DXA), and anthropometry (waist and hip circumference). Secondary outcome measures included fasting plasma glucose and insulin as well as fasting serum triglycerides, total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol. Higher physical activity energy expenditures were significantly associated with lower serum CRP levels (r = -0.18, P = 0.041), independent of oral HRT use, age, smoking behavior, alcohol consumption, aspirin use, and statin use. However, in the complete multivariate model, which included body fat, older ages (P = 0.047), greater trunk fat masses (P < 0.001), any oral HRT use (P < 0.001), and unopposed oral estrogen use (P = 0.012) were the sole independent predictors of higher serum CRP levels. The complete multivariate model accounted for 58% of the variance in serum CRP. We conclude that the association between higher physical activity and lower serum CRP levels is dependent on the lower body fat of the more active women, yet independent of oral HRT use. Future intervention trials should determine whether diet- and exercise-related reductions in body fat may be effective ways to diminish the proinflammatory effects of oral HRT in postmenopausal women. PMID- 12552584 TI - Effect of the bubonic plague epidemic on inbreeding in 14th century Britain. AB - The purpose of the study reported here was to further investigate a new method of estimating inbreeding in large, relatively isolated populations over historic times, as reported by Pattison (1999, 2001). The method is based on modeling the genealogical "paradox" and produces values of Pearl's coefficients of inbreeding as a function of time for any given population curve. In particular, this study demonstrates that the new method of estimating inbreeding may be used to examine the effect of unusual features in population curves on levels of inbreeding in that population. As an example, the medieval population "bump" that spans the 13th and 14th centuries in the population of Britain, which terminated abruptly with the outbreak of bubonic plague in 1348 AD, is examined. It is first assumed that the whole population was the adult (breeding) population, corresponding to minimum inbreeding. For this case it is found that, for distant generations, about 90% of the whole population occurs in any present-day British pedigree. This value compares favorably with the values of 85% and 80% reported by Wachter (1980) and Derrida et al. (1999, 2000), respectively. The population bump causes the curves for Pearl's coefficient, for minimum inbreeding, to have a plateau of about 96% over the period from about 1220 AD to 1380 AD. The effect of introducing a single known estimate of inbreeding into the model is to produce a more realistic situation where the adult (breeding) population is considerably less than the whole population. It is found that the maximum number of different ancestors in any generation, in the average British pedigree, is about 22900 individuals and occurs about 1220 AD. More importantly, and possibly contrary to expectation, it is now found that the population bump had virtually no effect on the level of inbreeding. PMID- 12552585 TI - How sexually dimorphic are we? Review and synthesis. PMID- 12552586 TI - Treatment and sampling protocols for transgenic mutation assays. AB - The standard protocol for testing chemicals with the transgenic mutation assays in vivo includes a period of time between treatment and sampling to permit the mutation frequency to reach a maximum. Recent evidence has shown, however, that for some chemicals the mutant frequency can decline substantially during this period, which would reduce the sensitivity of the assay. Here we discuss alternate protocols to maintain the sensitivity of the assay for both types of mutagens and, in particular, propose that treatments should continue until the time of sampling. PMID- 12552587 TI - Development and characterization of CHO repair-proficient cell lines for comparative mutagenicity and metabolism of heterocyclic amines from cooked food. AB - In order to understand the role of repair and metabolism in the mutagenicity of heterocyclic amines from cooked foods, we previously developed the nucleotide excision repair-deficient CHO 5P3NAT2 cell line engineered to coexpress the mouse CYP1A2 and human N-acetyltransferase genes. In the present study, we have made a matched repair-competent cell line by mutagenizing 5P3NAT2 cells with ethyl methanesulfonate and selecting for resistance to cytotoxicity by 2-amino-3 methylimidazo[4,5-f]quinoline (IQ). In the differential cytotoxicity (DC) assay, 4 out of 15 clones showed no cytotoxic effect with IQ at the highest dose (30 microg/ml) tested, in contrast to repair-deficient 5P3NAT2 cells, which showed approximately 100% cytotoxicity at 0.3 microg/ml. Subsequently, these IQ resistant clones were examined for resistance to killing by UV irradiation. All four IQ-resistant clones, which show resistance to UV similar to that of repair proficient AA8 cells, still express both the CYP1A2 and N-acetyltransferase genes. Sequence analysis of CXPD cDNA from the 5P3NAT2R9 clone revealed an A:T- >G:C reversion event at the site of the UV5 mutation. This base change results in reversion of the codon 116 tyrosine in UV5 cells back to the original cysteine in AA8 cells, thereby restoring wild-type CXPD activity and repair function. In contrast to 5P3NAT2 cells, the repair-proficient 5P3NAT2R9 revertant cell line shows little IQ-induced cell killing, and dramatically lower levels of induced mutation at the adenine phosphoribosyltransferase (Aprt) gene locus over the range of 2-40 microg/ml IQ. This matched pair of repair-proficient/deficient cell lines can provide insight not only into the genotoxicity of heterocyclic amine dietary carcinogens such as IQ and PhIP, but also into the effects of nucleotide excision repair on the ultimate mutagenicity of these compounds. PMID- 12552588 TI - Using CTLL-2 and CTLL-2 bcl2 cells to avoid interference by apoptosis in the in vitro micronucleus test. AB - In vitro assays for chromosome aberrations (i.e., in vitro micronucleus and in vitro metaphase analysis tests) frequently produce false-positive or exaggerated positive results. Our previous work suggested that apoptosis interferes with these tests, producing misleading results. These previous studies were conducted by performing the in vitro micronucleus test in CTLL-2 cells and a CTLL-2 cell derivative stably transfected with the apoptosis inhibitor gene bcl2. In the present study, these previous observations were extended by examining micronucleus induction with a larger number of compounds in both CTLL-2 and CTLL 2 bcl2 cells and measuring apoptosis with annexin V-FITC. Both cell lines were treated with different classes of compounds that were anticipated to be exclusively apoptosis inducers, or compounds known to be clastogens or aneugens, some of which were anticipated to be both genotoxic and apoptotic. We were able to confirm that compounds that are only apoptogenic induced micronuclei in CTLL-2 but not CTLL-2 bcl2 cells, indicating that the positive responses are due to apoptosis in CTLL-2 cells. Some genotoxins (clastogens and aneugens) did not produce apoptosis by the annexin V assay and gave similar responses in CTLL-2 and CTLL-2 bcl2 cells. Finally, higher responses were induced in CTLL-2 cells compared to CTLL-2 bcl2 cells that were treated with aneugens or clastogens that were also apoptosis inducers, suggesting that the greater response in CTLL-2 cells is a consequence of both genotoxicity and apoptosis. Finally, it was demonstrated that just eliminating CTLL-2 cells having three or more micronuclei from scoring was not adequate for correctly evaluating agents that only produce apoptosis. The results indicate that coupling the in vitro micronucleus test in both CTLL-2 and CTLL-2 bcl2 cells with the measurement of apoptosis is able to distinguish the genotoxic effects of a test compound from its apoptotic potential and is able to avoid interference from apoptosis in the in vitro micronucleus test. These observations may provide the basis for a useful genotoxicity assay. PMID- 12552589 TI - Molecular and cellular influences of butylated hydroxyanisole on Chinese hamster V79 cells treated with N-methyl-N'-nitro-N-nitrosoguanidine: antimutagenicity of butylated hydroxyanisole. AB - The antioxidant butylated hydroxyanisole (BHA) is a rodent carcinogen that also reduces the mutagenicity and carcinogenicity of other agents. In this study, we have evaluated possible mechanisms for the antimutagenicity of BHA by investigating its effects on N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-treated Chinese hamster V79 cells. Mutant frequency was determined using the hprt/V79 assay, while plating efficiency was used to measure cytotoxicity, and apoptosis was measured by flow immunofluorocytometry. In addition, DNA strand breaks and the kinetics of strand-break rejoining were investigated by the alkaline elution of DNA and by single-cell gel electrophoresis (SCGE). Although the higher concentration of BHA (0.5 mM) increased the cytotoxicity of MNNG and the lower concentration of BHA (0.25 mM) did not change it, both concentrations were antimutagenic in MNNG-treated cells, with the greater effect occurring at the lower BHA concentration. Neither BHA nor MNNG nor BHA + MNNG increased the level of apoptotic nuclei, and BHA did not change the level of MNNG-induced DNA strand breaks, though it did inhibit their rejoining. Determination of O(6) methylguanine-DNA-methyltransferase (MGMT) activity confirmed that V79 cells do not synthesize active MGMT protein; MGMT activity was also undetectable after MNNG and BHA + MNNG treatment. The ability of BHA to reduce the level of MNNG induced mutations did not correlate with cytotoxicity, induction of apoptosis, the level of DNA strand break induction, or MGMT activity. A modified SCGE assay showed that BHA significantly reduced the level of formamidopyrimidine-DNA glycosylase + endonucleaseIII-sensitive sites, which at least partially are caused by oxidative DNA lesions. The results suggest that the protective effect of BHA on MNNG-induced mutagenicity is best explained by the antioxidative activity of BHA, which may scavenge free radicals that participate in MNNG induced mutagenicity. PMID- 12552590 TI - Influence of common XPD and XRCC1 variant alleles on p53 mutations in lung tumors. AB - The DNA repair proteins XPD and XRCC1 are involved in the nucleotide and base excision repair of DNA lesions induced by many tobacco and environmental carcinogens. Common variant alleles at the XPD (312Asn, 751Gln) and XRCC1 (399Gln) loci have been identified and associated with increased risk for lung cancer. We therefore investigated a possible effect of these variant alleles on the frequency and spectrum of p53 mutations in the tumors of 97 Swedish lung cancer patients (56 never-smokers and 41 age-, gender-, and hospital-matched ever smokers). The p53 gene was mutated in 4 never-smokers (7%) and 11 ever-smokers (27%). Smoking-related transversion-type mutations predominated over transitions among smokers (8:3), but not among never-smokers (1:3). None of the variant alleles altered the overall frequency of p53 mutation. Transversions, however, were marginally increased among patients with at least one XPD variant allele compared with patients who were wild-type homozygotes (73% vs. 25% for the Asp312Asn polymorphism, P = 0.095; 78% vs. 33% for Lys751Gln, P = 0.085). Five of six women or six of seven smokers who carried at least one XPD 751Gln allele had p53 transversion. The XRCC1 variant allele did not show any effect on the p53 mutation. We conclude that the XPD variant alleles may be associated with an increased frequency of smoking-related p53 mutations in lung tumors, presumably due to reduced DNA repair proficiency. PMID- 12552591 TI - Genotoxicity and toxicity of the potential cancer-preventive agent polyphenon E. AB - The potential health benefits of green tea continue to attract public and scientific interests and are attributed in part to polyphenolic catechin constituents. Polyphenon E (Poly E) is a decaffeinated green tea catechin mixture containing about 50% epigallocatechin gallate and 30% other catechins. We evaluated the toxicity and genotoxicity of Poly E by using two in vitro assays: bacterial mutagenesis in a Salmonella typhimurium-E. coli assay and the L5178Y mouse lymphoma cell thymidine kinase (Tk) gene mutation assay. In addition, we used two in vivo genotoxicity assays: the mouse micronucleus assay and the Big Blue cII transgenic mouse mutation assay. Repeat-dose toxicity evaluations were performed in mice in parallel with the Big Blue transgenic mutation assays. No significant increases in the revertant colonies were found in the bacterial mutagenesis assay, but a significant increase in the mutant frequency (MF) at the Tk locus was observed in the mouse lymphoma test system. We observed toxicity in mice when Poly E was administered at doses of 2,000 mg/kg/day. Lower doses produced no significant increases in micronucleated erythrocytes in the bone marrow of Swiss-Webster mice and no significant increases in cII transgene MF in the liver, lung, or spleen compared with controls. These results indicate that Poly E, although toxic at high doses (2,000 mg/kg/day), poses minimal genotoxic concern. In addition, these studies highlight the importance of using both in vitro and in vivo systems in genetic toxicity screening of pharmaceuticals before they are administered to humans. PMID- 12552592 TI - Mutagenic activation of arylamines by subcellular fractions of Chamaelea gallina clams exposed to environmental pollutants. AB - Biochemical measurements in the sentinel clam Chamaelea gallina have been used as biomarkers of marine pollution. In this study, S9, cytosolic fractions (CF), and microsomal fractions (MF) prepared from unexposed clams and clams exposed to model pollutants were used to activate 2-aminoanthracene (2-AA) and 2 acetylaminofluorene (AAF) to mutagens in Salmonella typhimurium strain BA149, which overexpresses O-acetyltransferase. Arylamine activation was similar with subcellular fractions from unexposed and Aroclor 1254-exposed clams, but decreased with fractions from As(III)- and Cu(II)-exposed clams. Bioactivation of arylamines by CF was higher than by MF, and higher with NADH than with NADPH as the reducing agent. alpha-Naphthoflavone inhibited AAF activation by CF and MF, but increased 2-AA activation nearly twofold. In contrast to the results with arylamine activation, benzo[a]pyrene hydroxylase (BPH) activity increased twofold in fractions from Aroclor 1254- and Cu(II)-exposed clams. Activation of 2-AA was also evaluated using S9 fractions from clams sampled at littoral sites with different pollutant levels. Compared to a reference site, lower 2-AA bioactivation and higher BPH activity were found in clams containing high levels of copper and organic contaminants, although the differences were not statistically significant. While these findings agree with the results of the model Cu(II) exposure, the effects of other pollutants cannot be ruled out. The results of the study demonstrate that arylamine activation by clams is not preferentially catalyzed by microsomal monooxygenases but by unknown cytosolic system(s), and that bioactivation of 2-AA and AAF appears to occur by different pathways. PMID- 12552593 TI - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced lipid peroxidation and DNA damage in mouse bone marrow and blood. AB - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a neurotoxin that induces Parkinsonism in humans, monkeys, and mice and oxidative stress in mammalian cells and tissues. In the present study, the relationship between the generation of lipid peroxidation products and DNA damage was studied in mice treated with MPTP. The frequency of micronucleated polychromatic erythrocytes (MN-PCE) and the concentrations of malonaldehyde and 4-hydroxyalkenals were determined in the bone marrow and peripheral blood of mice 0, 24, 48, 72, and 96 hr after treatment with MPTP, cyclophosphamide as a positive control, or diluent. Both MN-PCE and the lipid peroxidation products increased in MPTP-treated mice, with significant levels being detected in bone marrow starting at 24 hr after treatment and in blood starting at 48 hr after treatment. These results suggest that the generation of oxidative products is related to the DNA damage produced by MPTP in mice. PMID- 12552594 TI - Polymorphisms for chemical metabolizing genes and risk for cervical neoplasia. AB - Infection with high-risk human papillomavirus (HPV) plays a major role in the etiology of cervical cancer (CC). However, most infected women do not develop cancer. Therefore, exposure to other carcinogenic agents may be a contributing risk factor for CC. We investigated the hypothesis that environmental exposure to cigarette smoke and inheritance of polymorphic chemical metabolizing genes (CYP2E1, GSTM1, and mEH) significantly increase the risk for neoplasia. We selected 76 cases with high-grade cervical neoplasia or with invasive CC and 75 matched healthy controls. The collected data support the well-established observation that infection with high-risk HPV is the major risk factor for CC (OR = 75; 95% CI = 26-220). In addition, our data show that women who smoked more than 15 "pack-year" had a significant 6.9-fold increase in risk (95% CI = 1.2 40.3) after adjustment for HPV infection. The CYP2E1 variant genotype did not significantly increase the risk for neoplasia. A significant increase in risk for neoplasia was observed for the low-activity mEH 113 His allele after adjustment for smoking (OR = 3.0; 95% CI = 1.4-6.3). The GSTM1 null genotype was associated with a significant 3.3-fold increased risk for neoplasia (95% CI = 1.0-11.8) compared to women who were GSTM1-positive after adjustment for smoking and HPV infection. Our study suggests that genetic differences in the metabolism of cigarette smoke, particularly GSTM1, may confer susceptibility to CC. Further studies using larger populations will be needed to confirm our observations and to validate data for disease prevention. PMID- 12552596 TI - Radionuclide imaging in the post-genomic era. AB - The assessment of gene function, which follows the completion of human genome sequencing, may be performed using the tools of the genome program. These tools represent high-throughput methods evaluating changes in the expression of many or all genes of an organism at the same time in order to investigate genetic pathways for normal development and disease. They describe proteins on a proteome wide scale, thereby, creating a new way of doing cell research which results in the determination of three dimensional protein structures and the description of protein networks. These descriptions may then be used for the design of new hypotheses and experiments in the traditional physiological, biochemical, and pharmacological sense. The evaluation of genetically manipulated animals or new designed biomolecules will require a thorough understanding of physiology, biochemistry, and pharmacology and the experimental approaches will involve many new technologies including in vivo imaging with SPECT and positron emission tomography (PET). Nuclear medicine procedures may be applied for the determination of gene function and regulation using established and new tracers or using in vivo reporter genes such as genes encoding enzymes, receptors, antigens, or transporters. Pharmacogenomics will identify new surrogate markers for therapy monitoring which may represent potential new tracers for imaging. Also drug distribution studies for new therapeutic biomolecules are needed at least during preclinical stages of drug development. Finally, new biomolecules will be developed by bioengineering methods, which may be used for isotope-based diagnosis and treatment of disease. PMID- 12552597 TI - Functional and molecular MR imaging of angiogenesis: seeing the target, seeing it work. AB - Intensive research over the last years led to the discovery of multiple molecular pathways and intricate regulatory network controlling the growth and regression of blood vessels in general and angiogenesis in particular. The difficulties in elucidation of the regulation of angiogenesis, stems from the inherent complexity due to participation of many cell types, under a dominant impact of physiological and environmental effects of flow, perfusion, and oxygenation. Major advances were achieved with the use of sophisticated transgenic mice models engineered so as to provide spatially and temporally controlled expression of specific factors alone or in combination. In vivo analysis of these models frequently requires the use of non-invasive imaging modalities for measurement of functional parameters of the vasculature along with dynamic molecular information. Optical methods are extensively applied for the study of angiogenesis [Brown et al., 2001] but provide very limited tissue penetration. MRI offers the advantage of being non invasive with uniform and relatively high spatial resolution for deep tissues. Multiple MRI approaches for monitoring angiogenesis were developed over the last years, each looking at a particular step in the process. The aim of this paper is to analyze the clinical, pharmaceutical, and biological needs for imaging of angiogenesis, and to critically evaluate the strengths and weaknesses of functional and molecular imaging for monitoring angiogenesis. The inherent problem of validation of different measures of angiogenesis, and the advantages and limitations associated with application of MRI based methods, as surrogates for other measurements of angiogenesis will be discussed. The terms molecular imaging and functional imaging are frequently loosely defined with a significant overlap between the two. For the sake of this paper we will apply a narrower definition of both terms, where molecular imaging will apply to methods directed towards detection of specific biological molecules that participate directly in (regulation of) a physiological process; while functional imaging will be used to describe those methods that aim to detect the physiological response to a defined (molecular) stimulus. PMID- 12552598 TI - Observations on the role of nuclear medicine in molecular imaging. AB - The phrase "molecular imaging" is unquestionably current and is receiving ever increasing use. For example, two organizations, the Institute for Molecular Imaging and the Academy of Molecular Imaging have recently been established with molecular imaging as their focus, with journal entitled "Molecular Imaging" and "Molecular Imaging and Biology," respectively. Furthermore, the two leading journals in the field of nuclear medicine have recently added this phrase to their covers-becoming the "European Journal of Nuclear Medicine and Molecular Imaging" and "The Journal of Nuclear Medicine-advancing molecular imaging." The National Institute of Biomedical Imaging and Bioengineering is the newest institute of the NIH. With this degree of attention, it may be surprising that there is as yet no universally accepted definition of molecular imaging. Numerous diverse definitions, some quite complex, have been proposed. With some exceptions, they all refer to imaging in the living animal of function at the cellular or molecular level. Thus molecular imaging may be defined as the observation of biological function at the molecular level in health and disease through some process involving non-invasive imaging of the living mammals. PMID- 12552599 TI - Molecular imaging of regional brain tumor biology. AB - Energy metabolism measurements in gliomas in vivo are now performed widely with positron emission tomography (PET). This capability has developed from a large number of basic and clinical science investigations that have cross fertilized one another. This article presents several areas that exemplify questions that have been explored over the last two decades. While the application of PET with [(18)F]-2-fluoro-2-deoxyglucose (FDG-PET) has proven useful for grading and prognosis assessments, this approach is less clinically suitable for assessing response to therapy, even though results to date raise very intriguing biological questions. Integration of metabolic imaging results into glioma therapy protocols is a recent and only preliminarily tapped method that may prove useful in additional trials that target DNA or membrane biosynthesis, or resistance mechanisms such as hypoxia. There are exciting future directions for molecular imaging that will undoubtedly be fruitful to explore, especially apoptosis, angiogenesis and expression of mutations of genes, e.g., epidermal growth factor receptor, that promote or suppress cellular malignant behavior. PMID- 12552600 TI - Non-invasive imaging of reporter genes. AB - Non-invasive, quantitative and repetitive imaging of biological processes in living animals is rapidly changing the way in which many experiments in models of human disease and normal physiological processes are conducted. This review summarizes the newest molecular imaging approaches to analyzing reporter gene expression, with particular emphasis on pre-clinical cancer research. Alternative modes of imaging are summarized, followed by descriptions of the major reporter gene systems now used for radionuclide imaging in vivo of gene expression. Several somatic delivery paradigms for co-ordinate expression of therapeutic and imaging genes are presented, and our own emphasis on the dopamine D2 receptor and Herpes Simplex Virus Type I thymidine kinase reporter genes are detailed. PMID- 12552601 TI - Quantitative assessment of tumor oxygen dynamics: molecular imaging for prognostic radiology. AB - One of the fundamental molecules governing the survival of mammalian cells is oxygen. Oxygen has gained particular significance in tumor developmental biology and oncology. An increasingly diverse array of methods is now available to characterize tumor oxygenation. This Prospect will consider a new method, Fluorocarbon Relaxometry using Echo planar imaging for Dynamic Oxygen Mapping (FREDOM), which we have recently developed for oximetry, examine application to a specific therapeutic example and place this technique in the context of other approaches. PMID- 12552602 TI - Fluorescence emission-based detection and diagnosis of malignancy. AB - Over the past decades, laser use in medicine has expanded from its initial application as a light-based scalpel to a plethora of clinical uses, ranging from surgical treatment through composite polymerization, dental ablation, vision correction, and skin resurfacing to diverse diagnostic modalities. Recently, the concept of light-based diagnostics and therapy has come under investigation. Low light intensities are used to excite endogenous or exogenous fluorophores, some of which have characteristic fluorescence emissions in pathological tissues. Thus, premalignancy and malignancy potentially can be detected and diagnosed. Photosensitized superficial lesions can subsequently be destroyed selectively by using higher intensities of laser light. The application of fluorescence emission based detection and diagnosis of precancer and cancer is reviewed, based on its application to the oral cavity-the author's primary anatomical area of expertise. This approach is justified as the same principles apply throughout the human body; to any area accessible to the clinician either directly or by some sort of fiber-optic probe. PMID- 12552603 TI - Evolution of radiopharmaceuticals for diagnosis and therapy. AB - Diagnostic radiopharmaceuticals should reflect in vivo biochemistry. Therapeutic radiopharmaceuticals should kill tumor cells while sparing healthy cells. As the understanding of a biological process increases, synthetic chemists must be ever poised to improve the design of new radiotracers to reflect the in vivo scenario in an accurate and precise fashion. However, it is somewhat ironic that, due to genetic variability, there is an inverse relationship between the specificity of the radiotracer and the number of individuals who will eventually be helped by these discoveries. Regardless of the beauty of the science that underpins radiopharmaceutical development, the availability of financial resources will direct the overall progress in the field. PMID- 12552604 TI - Combined vascular targeted imaging and therapy: a paradigm for personalized treatment. AB - In order to be successful in personalizing treatment, methods for selecting patients as well as good surrogate biomarkers for monitoring the effects of treatment are required in addition to development of an efficacious targeted therapy. We have developed a polymerized nanoparticle platform technology that will allow us to put different targeting moieties on the surface of the particles in addition to loading the particles with different contrast and therapeutic agents. We have proven that these nanoparticles can be targeted to endothelial receptors and different payloads of contrast and therapeutic agents have been delivered to target cells with high target to background ratios. Using this combined vascular targeted imaging and therapy approach, we are optimistic that personalized treatment regimens can be developed for different disease processes such as cancer, inflammation, and ischemia. PMID- 12552605 TI - Molecular imaging of tumor angiogenesis. AB - The emergence of angiogenesis as an important target for cancer therapy has led to increased research aimed at understanding the mechanisms underlying the development, maintenance, and destruction of tumor vasculature. Concurrently, molecular imaging technologies have been developed and are being incorporated as integral components of biomedical research due to their ability to noninvasively monitor in vivo molecular events. With the evaluation of numerous anti-angiogenic agents in clinical trials, the adaptation and validation of molecular imaging modalities for monitoring angiogenesis is actively being pursued. The importance of selecting appropriate molecular targets in the study of angiogenesis has become increasingly complex due to the pleiotropy of vascular phenotypes. Furthermore, due to both the relatively low abundance of endothelial cells in tumor tissue and the inherent difficulties of detecting molecular events, molecular imaging of vasculature necessitates continued improvements in achieving higher sensitivity. While several studies have been published that set the groundwork for imaging angiogenesis, much has yet to be accomplished. Various tumor models and transgenic mice provide an excellent resource for developing molecular imaging technologies for the understanding of angiogenesis. This research may play a particularly crucial role in evaluating mechanism and efficacy during pre-clinical testing of anti-angiogenic drugs. Due to practical limitations, however, the implementation of angiogenesis-directed molecular imaging may not extend beyond highly specialized clinical trials. That is, imaging modalities that evaluate angiogenesis at a functional level may prove more appropriate. Despite future technical challenges, molecular imaging will become an important research and clinical tool in evaluating tumor angiogenesis. PMID- 12552606 TI - Chemical microarrays, fragment diversity, label-free imaging by plasmon resonance -a chemical genomics approach. AB - Chemical genomics aim to create synergy between synthetic small molecule chemistry and biosciences employing genomic tools and information. Central to chemical genomics is the discovery of bioactive compounds from novel targets for pharmaceutical lead development. The field is challenged both by the multitude and novelty of protein and other biomacromolecular targets to be studied. Affinity fingerprints, data sets of binding interactions between collections of chemicals and their macromolecular receptors, hold promise to guide drug design and study protein function for groups of related compounds and families of biomacromolecules. Despite their fundamental relevance, neither experimental protocols nor databases of quantitative and comprehensive description of binding interactions for small molecule ligands and biomacromolecular receptors are available. Chemical microarrays in combination with label-free imaging provide a novel route towards the systematic and standardized acquisition and application of such affinity fingerprint information. PMID- 12552607 TI - Molecular imaging: an old and new field connecting basic science and clinical medicine. AB - Combination of recent progress in imaging technology and molecular biology/gene technology has evolved a new field named "molecular imaging". It includes wide range of imaging technique from basic research to clinical practice. For basic researchers, we focused on the part of in vivo imaging in human, and introduce the recent progress of modern biomedical imaging, as a crossing point of basic science and clinical medicine. PMID- 12552608 TI - Molecular imaging, targeted therapeutics, and nanoscience. AB - The recent emergence of "molecular imaging" as an academic discipline has set the stage for an evolutionary leap in diagnostic imaging. Recent advances in nuclear, ultrasound, optical, and magnetic resonance imaging have generated interest in molecular imaging across all modalities and across various academic, industrial, and governmental agencies. In this perspective, examples of the progress and the prospects for the future of molecular imaging and linked targeted therapeutics are reviewed. PMID- 12552609 TI - Prospects of molecular imaging in neurology. AB - Molecular imaging aims towards the non-invasive kinetic and quantitative assessment and localization of biological processes of normal and diseased cells in vivo in animal models and humans. Due to technological advances during the past years, imaging of molecular processes is a rapidly growing field, which has the potential of broad applications in the study of cell biology, biochemistry, gene/protein function and regulation, signal transduction, characterization of transgenic animals, development of new treatment strategies (gene or cell-based) and their successful implementation into clinical application. Most importantly, the possibility to study these parameters in the same subject repeatedly over time makes molecular imaging an attractive technology to obtain reliable data and to safe recourse; for example, molecular imaging enables the assessment of an exogenously introduced therapeutic gene and the related alterations of endogenously regulated gene functions directly in the same subject. Therefore, molecular imaging will have great implications especially when molecular diagnostic and treatment modalities have to be translated from experimental into clinical application. Here, we review the three main imaging technologies, which have been developed for studying molecular processes in vivo, the disease models, which have been studied so far, and the potential future applications. PMID- 12552610 TI - Molecular imaging: the application of small animal positron emission tomography. AB - The extraordinary advances in genomic technologies over the last decade have led to the establishment of new animal models of disease. The use of molecular imaging techniques to examine these models, preferably with non-destructive imaging procedures, such as those offered by positron emission tomography (PET), are especially valuable for the timely advancement of research. With the use of small animal PET imaging it is possible to follow individual subjects of a sample population over an extended time period by using highly specific molecular probes and radiopharmaceuticals. In this Prospect small animal PET imaging will be described, specifically focusing on the current technologies, its applications in molecular imaging and the logistics of performing small animal PET. PMID- 12552611 TI - Molecular imaging in small animals--roles for micro-CT. AB - X-ray micro-CT is currently used primarily to generate 3D images of micro architecture (and the function that can be deduced from it) and the regional distribution of administered radiopaque indicators, within intact rodent organs or biopsies from large animals and humans. Current use of X-ray micro-CT can be extended in three ways to increase the quantitative imaging of molecular transport and accumulation within such specimens. (1) By use of heavy elements, other than the usual iodine, attached to molecules of interest or to surrogates for those molecules. The accumulation of the indicator in the physiological compartments, and the transport to and from such compartments, can be quantitated from the imaged spatial distribution of these contrast agents. (2) The high spatial resolution of conventional X-ray attenuation-based CT images can be used to improve the quantitative nature of radionuclide-based tomographic images (SPECT & PET) by providing correction for attenuation of the emitted gamma rays and the accurate delineation of physiological spaces known to selectively accumulate those indicators. Similarly, other imaging modalities which also localize functions in 2D images (such as histological sections subsequently obtained from the same specimen), can provide a synergistic combination with CT based 3D microstructure. (3) By increasing the sensitivity and specificity of X ray CT image contrast by use of methods such as: K-edge subtraction imaging, X ray fluorescence imaging, imaging of the various types of scattered X-ray and the consequences of the change in the speed of X-rays through different tissues, such as refraction and phase shift. These other methods of X-ray imaging can increase contrast by more than an order of magnitude over that due to conventionally-used attenuation of X-ray. To fully exploit their potentials, much development of radiopaque indicators, scanner hardware and image reconstruction and analysis software will be needed. PMID- 12552612 TI - Model-based biological Raman spectral imaging. AB - Raman spectral imaging is a powerful tool for determining chemical information in a biological specimen. The challenge is to condense the large amount of spectral information into an easily visualized form with high information content. Researchers have applied a range of techniques, from peak-height ratios to sophisticated models, to produce interpretable Raman images. The purpose of this article is to review some of the more common imaging approaches, in particular principal components analysis, multivariate curve resolution, and Euclidean distance, as well as to present a new technique, morphological modeling. How to best extract meaningful chemical information using each imaging approach will be discussed and examples of images produced with each will be shown. PMID- 12552614 TI - Magnetic resonance microscopy approaches to molecular imaging: sensitivity vs. specificity. AB - Magnetic resonance imaging (MRI) has become a staple of diagnostic radiology. Despite its diagnostic utility the resolving power of typical clinical MRI instruments is only on the order of 1 mm. This has led to the development of magnetic resonance microscopy (MRM), which employs the same physical imaging principals used in MRI, but with instrumentation designed to resolve structural details down to the level of 10-100 microns in samples ranging from less than 1mm to several centimeters in size. Until recently, major advancements in MRM have focused on hardware and software developments allowing the detection of radio frequency signals originating from very small volume elements within the sample. Such high-resolution images have facilitated the early detection of diseased tissue by focusing on sub-millimeter structural changes induced in the tissue. To sensitize the MRM technique to pathologic tissue changes, investigators have developed techniques, such as chemical shift imaging to detect pre-cancerous changes in tissue metabolism and MR relaxometry to detect changes in tissue composition during the earliest stages of degeneration for diseases such as osteoarthritis or multiple sclerosis. However, such non-specific measurements can only serve as surrogate measures of disease progression and potential measures of treatment efficacy. As disease diagnosis moves from the anatomic to the molecular stage, scientists will require imaging techniques that can detect molecular events deep inside the human body. To meet this goal, MR scientists are working to improve imaging resolutions in vivo and they are developing molecular probes that can dramatically amplify the MR signal in response to specific and highly localized molecular events. This article will identify current trends in the MRM field aimed at meeting the challenges imposed by molecular imaging and areas for future development in this highly promising imaging field. PMID- 12552613 TI - Non-invasive imaging of vulnerable plaques by molecular targeting of oxidized LDL with tagged oxidation-specific antibodies. AB - The concept of the "vulnerable" plaque has recently emerged to explain how quiescent atherosclerotic lesions evolve to cause clinical events. Vulnerable plaques are generally non-obstructive, asymptomatic lesions that may abruptly rupture and induce thrombotic occlusion leading to tissue ischemia and its attendant sequelae. They are responsible for over 50% of cases of sudden death and acute myocardial infarction. The lipid component of vulnerable plaques, which is abundant and highly enriched in oxidized low-density lipoprotein (OxLDL), strongly contributes to their propensity to rupture through physical, inflammatory, and thrombogenic properties. We hypothesized that OxLDL would serve as an ideal target to detect vulnerable plaques. In a series of experimental studies, we have shown that oxidation-specific antibodies (Ox-AB) specifically accumulate in vivo within lipid-rich atherosclerotic lesions but not normal arteries, provide a quantitative measure of the content of OxLDL, allow detection of atherosclerosis progression and regression in the context of enhanced or reduced OxLDL content and non-invasively image atherosclerotic lesions. Ox-AB may be tagged with appropriate labels for use in nuclear scintigraphy, magnetic resonance, or ultrasound imaging. Potential research and clinical applications include studying the natural history of atherosclerosis in animal models and humans, evaluating novel drug or genetic therapies on progression and regression of atherosclerosis, evaluating plaque stability, screening and serial follow-up of high-risk individuals, non-invasive imaging of vulnerable plaques, and assessing the clinical efficacy of new treatments of atherosclerosis. PMID- 12552615 TI - Nanobiosensors: probing the sanctuary of individual living cells. AB - Recently, nanotechnology has been revolutionizing important areas in molecular biology, especially diagnostics and therapy at the molecular and cellular level. The combination of nanotechnology, biology, and photonics opens the possibility of detecting and manipulating atoms and molecules using nanodevices, which have the potential for a wide variety of medical uses at the cellular level. The nanoprobes were fabricated with optical fibers pulled down to tips with distal ends having sizes of approximately 30-50 nm. The nanoscale size of this new class of sensors, allows for measurements in the smallest of environments. One such environment that has evoked a great deal of interest is that of individual cells. Using these nanobiosensors, it has become possible to probe individual chemical species in specific locations throughout a cell. This article provides an overview of the principle, development, and applications of optical nanosensor systems for in vivo bioanalysis at the single-cell level. The fiberoptics nanoprobes were covalently bound with antibodies that are selective to target analyte molecules. Excitation light is launched into the fiber and the resulting evanescent field at the tip of the fiber is used to excite target molecules bound to the antibody molecules. The fluorescence emission from the analyte molecules is then collected via a microscope. The usefulness and potential of this nanotechnology-based biosensor systems in biological research and applications in single-cell analysis are discussed. PMID- 12552616 TI - Molecular imaging: new applications for biochemistry. AB - Molecular imaging can reveal in vivo analysis and quantification of biochemical reactions. To enable cell-surface imaging of receptors, novel ligands have been developed which can be radiolabeled or imaged by bioluminescence. Specific examples include somatostatin receptors, estrogen and progesterone receptors, receptors involved in adhesion and externalization of phosphatidyl serine as an indicator of apoptosis. Central nervous system imaging can be carried out using ligands for receptors including dopamine, serotonin and Gamma amino butyric acid (GABA). In addition, tumor and metabolic imaging can be carried out with the Na-K ATPase pump using the tracer thallium-201 for SPECT or F-18 FDG for PET imaging. Finally, novel receptors or endogenous metabolic pathways can be analyzed combining cell-gene therapy to create specific tracer targets in cells that can be studied by molecular imaging. The challenge of molecular imaging is to first identify key pathways that are unique for a specific disease processes, such as atherosclerosis, cancer, CNS disorders, immunologic and arthritis disorders and next to devise a high-affinity specific small molecular ligand that can be adapted to be a radiolabeled tracer to study this pathway. Advances in genomics and proteomics combine with new peptide-chemistry approaches should provide a large number of targets and tracers in the near future to achieve these imaging objectives. PMID- 12552617 TI - In vivo molecular-genetic imaging. AB - Noninvasive in vivo molecular-genetic imaging has developed over the past decade and involves nuclear (PET, gamma camera), magnetic resonance, and in vivo optical imaging systems. Although three different imaging strategies - "direct," "indirect" and "surrogate" - are being used, most current in vivo molecular imaging strategies are "indirect" and involve the coupling of a "reporter gene" with a complimentary "reporter probe." Imaging the level of probe accumulation provides indirect information related to the level of reporter gene expression. Reporter gene constructs are driven by upstream promoter/enhancer elements; reporter gene expression can be "constitutive" leading to continuous transcription and used to identify the site of transduction and to monitor the level and duration of gene (vector) activity. Alternatively, reporter gene expression can be "inducible" leading to controlled gene expression. Controlled gene expression can be tissue-specific and/or responsive to the level of endogenous promoters and transcription factors. Several examples of imaging endogenous biological processes in animals using reporter constructs, radiolabeled probes and PET imaging are reviewed, including: 1) imaging transcriptional regulation (e.g., p53-dependent gene expression), 2) imaging weak promoters (cis- vs. trans-reporter configurations), 3) imaging post transcriptional regulation of gene expression, 4) imaging protein-protein interactions. The development of versatile and sensitive assays that do not require tissue sampling will be of considerable value for monitoring molecular genetic and cellular processes in animal models of human disease, as well as for studies in human subjects in the future. Non-invasive imaging of molecular genetic and cellular processes will compliment established ex vivo molecular biological assays that require tissue sampling, and will provide a spatial as well as a temporal dimension to our understanding of various diseases. PMID- 12552618 TI - Grappling with metastatic risk: bringing molecular imaging of Met expression toward clinical use. AB - The availability of a test to assess the likelihood that a given tumor will invade or metastasize would be a useful development in clinical oncology. We propose that multimodality imaging of tumor expression of Met could serve as a prototype for metastatic risk stratification (MRS). Met, a receptor protein tyrosine kinase, is expressed by most solid tumors, and aberrant expression of Met is associated with poor clinical prognosis. We summarize the current status and predict the future direction of research in four areas of molecular imaging and cancer therapy that exploit Met. PMID- 12552619 TI - Advances in molecular labeling, high throughput imaging and machine intelligence portend powerful functional cellular biochemistry tools. AB - Cellular behavior is complex. Successfully understanding systems at ever increasing complexity is fundamental to advances in modern science and unraveling the functional details of cellular behavior is no exception. We present a collection of prospectives to provide a glimpse of the techniques that will aid in collecting, managing and utilizing information on complex cellular processes via molecular imaging tools. These include: 1) visualizing intracellular protein activity with fluorescent markers, 2) high throughput (and automated) imaging of multilabeled cells in statistically significant numbers, and 3) machine intelligence to analyze subcellular image localization and pattern. Although not addressed here, the importance of combining cell-image-based information with detailed molecular structure and ligand-receptor binding models cannot be overlooked. Advanced molecular imaging techniques have the potential to impact cellular diagnostics for cancer screening, clinical correlations of tissue molecular patterns for cancer biology, and cellular molecular interactions for accelerating drug discovery. The goal of finally understanding all cellular components and behaviors will be achieved by advances in both instrumentation engineering (software and hardware) and molecular biochemistry. PMID- 12552620 TI - Can small animal imaging accelerate drug development? AB - Better mechanistic understanding of disease through mapping of the human and mouse genomes enables rethinking of human infirmity. In the case of cancer, for example, we may begin to associate disease states with their underlying genetic defects rather than with the organ system involved. That will enable more selective, nontoxic therapies in patients who are genetically predisposed to respond to them. Because one of the major goals of molecular imaging research is to interrogate gene expression noninvasively, it can impact greatly on that process. Most of molecular imaging research is undertaken in small animals, which provide a conduit between in vitro studies and human clinical imaging. We are fortunate to be able to manipulate small animals genetically, and to have increasingly better models of human disease. The ability to study those animals noninvasively and quantitatively with new, high-resolution imaging devices provides the most relevant milieu in which to find and examine new therapies. PMID- 12552621 TI - Introduction to the physics of molecular imaging with radioactive tracers in small animals. AB - Recent advances have greatly enhanced the three-dimensional (3D) imaging of radioactive tracers in living animals. this article introduces the physics of imaging behind the imaging methods. The article first discusses the selection of the radiation emitted from the tracer and then the process of tomographic reconstruction or how 3D images are made from imaging around the outside of the animal. The technique of single photon emission computed tomography (SPECT) in which the detection of one X-ray or gamma ray at a time is employed for image formation is then described. Finally, positron emission tomography (PET) which relies on the simultaneous detection of the pair of gamma-rays formed when the positron annihilates is presented. PMID- 12552622 TI - In vivo molecular imaging. AB - The relatively young field of molecular imaging is focused on the visualization of molecular phenotypes in whole organisms. This is achieved using imaging systems based on radionuclides, nuclear magnetic resonance, ultrasound, or the visible-IR region of the optical spectrum. Molecularly defined contrast in these modalities is generated by exogenous probes of the endogenous proteome, or through transgenes. Examples of exogenous probes include those that are transported and trapped (glucose, nucleoside analogs), those directed against extracellular receptors (somatostatin, opioid, melanotropin), and those activated by extracellular proteases. Transgenes that have been used in molecular imaging include the above receptors, non-mammalian enzymes that trap pro-drugs (HSV-tk, yeast CD), and optical reporter proteins (luciferase, fluorescent proteins). Cutting edge technologies in this field include in vivo assays for protein protein interactions, and in vivo assays for mRNA expression patterns. The number of degrees of freedom in designing new agents is daunting, and advancements in this field will require a significant participation from molecular and cellular biochemists. PMID- 12552623 TI - Understanding immune cell trafficking patterns via in vivo bioluminescence imaging. AB - Cell migration is a key aspect of the development of the immune system and mediating an immune response. There is extensive and continual redistribution of cells to different anatomic sites throughout the body. These trafficking patterns control immune function, tissue regeneration, and host responses to insult. The ability to monitor the fate and function of cells, therefore, is imperative to both understanding the role of specific cells in disease processes and to devising rational therapeutic strategies. Determining the fate of immune cells and understanding the functional changes associated with migration and proliferation require effective means of obtaining in vivo measurements in the context of intact organ systems. A variety of imaging methods are available to provide structural information, such as X-ray CT and MRI, but only recently new tools have been developed that reveal cellular and molecular changes as they occur within living animals. We have pioneered one of these techniques that is based on the observations that light passes through mammalian tissues, and that luciferases can serve as internal biological sources of light in the living body. This method, called in vivo bioluminescence imaging, is a rapid and noninvasive functional imaging method that employs light-emitting reporters and external photon detection to follow biological processes in living animals in real time. This imaging strategy enables the studies of trafficking patterns for a variety of cell types in live animal models of human biology and disease. Using this approach we have elucidated the spatiotemporal trafficking patterns of lymphocytes within the body. In models of autoimmune disease we have used the migration of "pathogenic" immune cells to diseased tissues as a means to locally deliver and express therapeutic proteins. Similarly, we have determined the tempo of NK-T cell migration to neoplastic lesions and measured their life span in vivo. Using bioluminescence imaging individual groups of animals can be followed over time significantly reducing the number of animals per experiment, and improving the statistical significance of a study since changes in a given population can be studied over time. Such rapid assays that reveal cell fates in vivo will increase our basic understanding of the molecular signals that control these migratory pathways and will substantially speed up the development and evaluation of therapies. PMID- 12552626 TI - Integrating self-help books into psychotherapy. AB - This article describes a systematic and integral method of incorporating self help books into psychotherapy as a collaborative function. We address the distinctions between self-help and bibliotherapy, consider bibliotherapy as adjunctive or integrative to psychotherapy, and outline the multiple uses of bibliotherapy for clinical purposes. How to apply self-help books in psychotherapy and ways to select books are illustrated by a case example. Indications and contraindications for bibliotherapy in therapy are outlined. PMID- 12552627 TI - Bibliotherapy as an adjunct to psychotherapy for depression in older adults. AB - Bibliotherapy, reading a self-help book for the treatment of psychological problems, has been shown to be effective as a "stand-alone" treatment for depression. Many practitioners recommend self-help books as an adjunct to treatment. This article offers some guidelines for the use of bibliotherapy as an adjunct to individual psychotherapy with depressed older adults. Two clinical cases demonstrate how bibliotherapy can be used effectively in conjunction with individual psychotherapy. PMID- 12552628 TI - The use of autobiography in psychotherapy. AB - First-person narratives may have advantages as adjuncts in psychotherapy. They provide an inside view of mental disorders expressed in the person's own words, emphasize issues that the person deems important, are interesting to read with strong story lines, are less didactic than self-help books, and offer identification with a protagonist. Recent trends in published autobiographies are described. There has been an increase in the number of published autobiographies describing mood disorder relative to schizophrenia, of psychotherapists going public with personal experiences of mental disorder, and of books expressing a positive view of treatment. The article includes case vignettes and a practitioner-recommended list of autobiographies on addictive disorders, death and grieving, and mood disorders. PMID- 12552629 TI - The clinical use of films in psychotherapy. AB - This article discusses the ways in which client attitudes about mental illness, psychotherapy, and therapists are shaped by contemporary films. Five common myths about mental illness that are promulgated by films are discussed, and the potential applications of films in psychotherapy are reviewed. Numerous examples of films relevant to psychotherapy are presented, and a clinical vignette is used to demonstrate how films can enrich and expand psychotherapy. PMID- 12552630 TI - Finding, evaluating, and incorporating Internet self-help resources into psychotherapy practice. AB - The Internet can be a wondrously rich source of information and support for clients. This article offers advice on how to make a practitioner's Web site valuable to potential clients, how to find Web sites of use to clients for self help, how to use Web sites in support of therapeutic efforts, how to evaluate the quality of Web sites, and finally, a case example of using the Internet with a multiproblem client. PMID- 12552631 TI - Focused expressive writing as self-help for stress and trauma. AB - In the therapy process, the process of disclosing about stressful or traumatic events is often considered essential. One such manner is through focused expressive writing (FEW) about stressful or traumatic experiences. FEW is related to improvements in health and well-being, across a wide array of outcomes and participant characteristics. As FEW requires limited involvement of other individuals, is relatively low cost, and portable, it has tremendous potential as self-help. In particular, FEW may be an effective means to reach populations unwilling or unable to engage in psychotherapy. A case illustration of FEW is presented. Evidence and future directions for FEW as self-help are reviewed. PMID- 12552632 TI - The effectiveness of self-administered treatments: a practice-friendly review of the research. AB - Self-administered treatments are a cost-effective way to treat a broad spectrum of people. This article focuses on the existing research of self-administered treatments and their effectiveness when integrated with ongoing practice or when implemented alone. Evidence for their effectiveness is mixed; self-help has been proven successful in the treatment of depression, mild alcohol abuse, and anxiety disorders. It has proven less successful for smoking cessation and moderate to severe alcohol abuse. When determining whether self-administered treatment is appropriate, individual characteristics and attitude as well as the nature and severity of the problem should be taken into consideration. In addition, because many self-help treatments have not been evaluated, caution should be exercised when implementing self-administered treatment, and progress should be carefully monitored. PMID- 12552633 TI - Unique expression patterns of the retinoblastoma (Rb) gene in intact and lens regeneration-undergoing newt eyes. AB - Based on the role of retinoblastoma (Rb) in lens development and in the cell cycle reentry of muscle cells during limb regeneration, we have analyzed expression or Rb patterns in intact and lens regeneration-undergoing newt eyes. We find that in intact newt eye Rb is expressed in the retina as a gradient with higher levels in the photoreceptor layer and virtually no expression in the ganglion layer. In addition, a second gradient was detected within the photoreceptor layer with expression diminishing at the dorsal and ventral regions. In the intact lens, Rb is expressed in the lens epithelium and in the differentiating lens fibers at the bow region. During lens regeneration, Rb is expressed very strongly in the differentiating lens fibers, but not in the lens epithelium. Using an antibody specific to the hyperphosphorylated form of Rb, we detected the inactive protein only in the pigment epithelial cells of the iris. These distinct patterns might be related to the regenerative potential of the lens in the newt. PMID- 12552635 TI - Embryonic origin of the caudal mesenteric artery in the mouse. AB - It is commonly held that the caudal mesenteric artery (CaMA, or inferior mesenteric artery in humans) arises in the same manner as the celiac and cranial mesenteric artery (CrMA, or superior mesenteric artery in humans), i.e., from the remodeling of the vitelline system of arteries that surrounds and supports the yolk sac. Conflicting evidence about the precise manner in which the CaMA arises was presented in studies of the luxate syndrome (Carter: J. Genet. 1954;52:1-35) and sirenomelia (Schreiner and Hoornbeek: J. Morphol. 1973;141:345-358) in the mouse. These studies suggested that the CaMA arises from the remodeling of the medial umbilical arterial roots. Later studies of blood vessel development in the hindlimb of the Dominant hemimelic mouse (Gest: Anat. Rec. 1984;208:296; Anat. Rec. 1987;218:49A; Gest and Roden: Anat. Rec. 1988;220:37-38A) also supported the results of the previous studies. The present investigation tests the hypothesis that the CaMA arises as a result of the regression and remodeling of the medial umbilical arterial roots. Vascular corrosion casts of 9.5-13.5-day-old mouse embryos were observed by scanning electron microscopy (SEM). The results of the present investigation agree with the aforementioned studies. The medial umbilical roots initially conduct the blood to the placenta. On days 10-12 the medial umbilical roots regress and remodel into the CaMA, while the lateral umbilical roots take over the blood supply to the placenta. On the basis of our results, we conclude that the CaMA arises from the medial umbilical roots and not from the remodeling of the vitelline system of arteries, as previously assumed. PMID- 12552634 TI - Slug expression during organogenesis in mice. AB - The vertebrate Slug gene encodes a zinc finger-containing transcriptional repressor. Here we report expression of the mouse Slug gene during organogenesis and late fetal development using histochemical detection of beta-galactosidase expressed from a targeted Slug(lacZ) knock-in allele. The Slug gene is highly expressed in the mesenchymal or stromal component of numerous organs. It is also highly expressed in craniofacial mesenchyme, in bone of both mesodermal and neural crest origin, and in the outflow tract and the endocardial cushions of the heart. PMID- 12552636 TI - VEGF modulates early heart valve formation. AB - Although hypoxic and/or nutritional insults during gestation are believed to contribute to congenital heart defects, the mechanisms responsible for these anomalies are not understood. Given the role vascular endothelial growth factor (VEGF) plays in response to hypoxia, it is a likely candidate for mediating deleterious effects of embryonic hypoxia. The ectopic or overproduction of endogenous factors such as VEGF may contribute to specific heart defects. Here we compared hypoxia-induced precocious production of VEGF during early heart valve development to normal VEGF production. Mouse prevalvular cardiac endocardial cushions were explanted onto hydrated type I collagen gels under normoxic or hypoxic conditions. The extent of transformation of cardiac endothelium into mesenchyme was inversely correlated with the levels of VEGF during the various culture conditions. A soluble VEGF antagonist confirmed that endogenous production of VEGF was specific for blocking normal cushion mesenchyme formation. We further demonstrated that E10.5 endocardium retains the ability to transform into cardiac mesenchyme in the absence of endogenous VEGF. PMID- 12552637 TI - Morphology and distribution of nitric oxide synthase-, neurokinin-1 receptor-, calretinin-, calbindin-, and neurofilament-M-immunoreactive neurons in the myenteric and submucosal plexuses of the rat small intestine. AB - Characterization of the enteric neurons is vital for understanding their physiological role. We have used single and dual label fluorescence and peroxidase-based immunohistochemistry in myenteric and submucosal whole mounts from the rat small intestine to evaluate the morphology and distribution of enteric neurons immunoreactive for the following phenotypic antigens: neuronal nitric oxide synthase (NOS), neurokinin-1 receptor (NK-1R), calretinin (Calr), calbindin (Cal), and neurofilament-M (NF-M). NOS-immunoreactive neurons had Dogiel type I morphology, were abundant in the myenteric plexus compared to the submucosal plexus, and never coexpressed NK-1R immunoreactivity. NK-1R- and Calr immunoreactive neurons had Dogiel type II morphology and were distributed comparably in both plexuses. NK-1R and Calr-immunoreactivity were coexpressed in many of the same neurons. Calbindin-immunoreactive neurons exhibited four distinct morphologies: small and large Dogiel type II neurons, Dogiel type I neurons, and small elongated neurons. These neurons were significantly fewer in number in the myenteric plexus compared to the submucosal plexus. Neurofilament-M immunoreactive neurons had three morphologies, Dogiel type II neurons, small Dogiel type II neurons, and a less common subpopulation of small, elongated, multipolar neurons. These neurons were also fewer in number in the myenteric plexus compared to the submucosal plexus. The distribution of these phenotypic markers may assist future work that elucidates the functional activities of these enteric neurons such as control of intestinal motility and adaptation to the entry of gastric contents. PMID- 12552638 TI - Bovine circumvallate taste buds: taste cell structure and immunoreactivity to alpha-gustducin. AB - The taste buds of bovine circumvallate papillae were investigated under light and electron microscopy both by histological and immunohistochemical methods. Taste buds existed in the inner epithelium of the trench of the papillae. Under electron microscopy, two types of taste cells, type I and type II, could be classified according to the existence of dense-cored vesicles and cytoplasmic density. Type I had electron-lucent cytoplasm and possessed many electron-dense cored vesicles in the apical cytoplasm. It was considered that the electron-dense materials of the vesicles were released and constituted the pore substance. This type of cell possessed long and thick apical processes in the taste pore. Type II had denser electron cytoplasm compared with that of type I and possessed many electron-lucent vesicles in the apical cytoplasm. This type of cell possessed microvilli in the taste pore. To know the immunoreactivity to alpha-gustducin in bovine circumvallate taste buds, we used the immunoblotting method and the immunohistochemical method. The alpha-gustducin reaction band at 40 kDa was displayed in the specimen of Western blots. The immunohistochemical property of the antiserum to alpha-gustducin was investigated by using the avidin-biotin complex (ABC) method and the 1.4-nm gold and silver enhancement methods. A subset of taste cells showed the immunoreactivity under light microscopy. The electron microscopic specimens with the 1.4-nm gold and silver enhancement method revealed that only type II cells exhibited the alpha-gustducin immunoreactivity. PMID- 12552639 TI - Friedrich Sigmund Merkel and his "Merkel cell", morphology, development, and physiology: review and new results. AB - Merkel nerve endings are mechanoreceptors in the mammalian skin. They consist of large, pale cells with lobulated nuclei forming synapse-like contacts with enlarged terminal endings of myelinated nerve fibers. They were first described by F.S. Merkel in 1875. They are found in the skin and in those parts of the mucosa derived from the ectoderm. In mammals (apart from man), the largest accumulation of Merkel nerve endings is found in whiskers. In all vertebrates, Merkel nerve endings are located in the basal layer of the epidermis, apart from birds, where they are located in the dermis. Cytoskeletal filaments consisting of cytokeratins and osmiophilic granules containing a variety of neuropeptides are found in Merkel cells. In anseriform birds, groups of cells resembling Merkel cells, with discoid nerve terminals between cells, form Grandry corpuscles. There has been controversy over the origin of Merkel cells. Results from chick/quail chimeras show that, in birds, Merkel cells are a subpopulation of cells derived from the neural crest, which thus excludes their development from the epidermis. Most recently, also in mammals, conclusive evidence for a neural crest origin of Merkel cells has been obtained. Merkel cells and nerve terminals form mechanoreceptors. Calcium ions enter Merkel cells in response to mechanical stimuli, a process which triggers the release of calcium from intracellular stores resulting in exocytosis of neurotransmitter or neuromodulator. Recent results suggest that there may be glutamatergic transmission between Merkel cell and nerve terminal, which appears to be essential for the characteristic slowly adapting response of these receptors during maintained mechanical stimuli. Thus, we are convinced that Merkel cells with associated nerve terminals function as mechanoreceptor cells. Cells in the skin with a similar appearance as Merkel cells, but without contact to nerve terminals, are probably part of a diffuse neuroendocrine system and do not function as mechanoreceptors. Probably these cells, rather than those acting as mechanoreceptors, are the origin of a highly malignant skin cancer called Merkel cell carcinoma. PMID- 12552640 TI - Distribution of vitamin A-storing lipid droplets in hepatic stellate cells in liver lobules--a comparative study. AB - To investigate the storage mechanisms of vitamin A, we examined the liver of adult polar bears and arctic foxes, which physiologically store a large amount of vitamin A, by high-performance liquid chromatography (HPLC), transmission electron microscopy (TEM) morphometry, gold chloride staining, fluorescence microscopy for the detection of autofluorescence of vitamin A, staining with hematoxylin-eosin (H&E), Masson's trichrome, and Ishii and Ishii's silver impregnation. HPLC revealed that the polar bears and arctic foxes contained 1.8 1.9 x 10(4) nmol total retinol (retinol plus retinyl esters) per gram liver. In the arctic foxes, the composition of the retinyl esters was found to be 51.1% palmitate, 26.6% oleate, 15.4% stearate, and 7% linoleate. The hepatic stellate cells of the arctic animals were demonstrated by TEM to contain the bulk of the vitamin A-lipid droplets in their cytoplasm. The liver lobules of the arctic animals showed a zonal gradient in the storage of vitamin A. The gradient was expressed as a symmetric crescendo-decrescendo profile starting at the periportal zone, peaking at the middle zone, and sloping down toward the central zone in the liver lobule. The density (i.e., cell number per area) of hepatic stellate cells was essentially the same among the zones. The gradient and the composition of the retinyl esters in storing vitamin A were not changed by differences in the vitamin A amount in the livers. These results indicate that the heterogeneity of vitamin A-storage capacity in hepatic stellate cells of arctic foxes and polar bears is genetically determined. PMID- 12552642 TI - Anatomical comparison of the cephalic musculature of some members of the superfamily Myliobatoidea (chondrichthyes): implications for evolutionary understanding. AB - This article describes the anatomy of the dorsal and ventral cephalic musculature of Gymnura marmorata, G. micrura, Aetobatus narinari, Myliobatis californica, M. longirostris, Rhinoptera steindachneri, Mobula munkiana, and M. thurstoni. It was observed that muscles of the dorsal cephalic region showed little variation among species, with the exception of the dorsal longitudinal bundles and the cucullaris muscle. The ventral cephalic musculature showed wider differences, mainly in the depressor hyomandibulae, coracomandibularis, and mandibular adductor muscles. M. munkiana and M. thurstoni revealed a significant muscle reduction, while M. californica, M. longirostris, A. narinari, and R. steindachneri showed a significant development of the ventral cephalic musculature. The species in this comparative study were clearly grouped based on their feeding habits. Data gathered on the muscle arrangements correspond to other taxonomy studies conducted on these groups. However, the results of this study agree only partially with those from previously described phylogenetic models. Therefore, further phylogenetic research is recommended. PMID- 12552643 TI - Annual household survey finds millions of Americans in denial about drug abuse. PMID- 12552641 TI - Porcine intrinsic cardiac ganglia. AB - The gross, light, and electron microscopic anatomies of the porcine intrinsic cardiac nervous system were investigated in 26 pigs to facilitate functional studies in this model. Gross anatomy: Numerous ganglia and interconnecting nerves (ganglionated plexuses) were found to be concentrated in epicardial fat in five atrial and six ventricular regions. The five atrial ganglionated plexuses identified were (1) the ventral right atrial, (2) the right vena cava-right atrial, (3) the dorsal atrial, (4) the interatrial septal, and (5) the left superior vena cava-left atrial ones. Six ventricular ganglionated plexuses were identified in close proximity to the (1) roots of the aorta and pulmonary artery (craniomedial), extending along the left main coronary artery to the (2) ventral interventricular and (3) circumflex coronary arteries. (4) A ganglionated plexus was identified around the origin of the dorsal interventricular coronary artery, as well as the (5) right main and (6) right marginal coronary arteries. Isolated neurons were identified scattered throughout the cranial interventricular septum. Microscopic anatomy: Approximately 3,000 neuronal somata were estimated to compose this intrinsic cardiac nervous system. Some ganglia contained more than 100 neurons. Neuronal somata had dimensions of roughly 33.1 (short axis) by 46.3 (long axis) microm. Most were multipolar, a small population of unipolar neurons being identified in atrial and ventricular tissues. At the electron microscopic level, asymmetrical axodendritic synapses with small clear, round vesicles were identified, some containing large dense-cored vesicles. In summary, porcine intrinsic cardiac neurons are concentrated in 11 distinct atrial and ventricular ganglionated plexuses. These extensive plexuses, along with fewer scattered neurons, display varied neuronal morphology and synaptology that represent the anatomical substrate for complex information processing within the intrinsic cardiac component of the porcine cardiac neuronal hierarchy. These anatomical data provide a framework for physiological analyses of the porcine intrinsic cardiac nervous system. PMID- 12552644 TI - Food safety: suggestions for improvement. PMID- 12552645 TI - Confronting chronic neglect. PMID- 12552646 TI - [Surface modification of low temperature isotropic pyrolytic carbon artificial heart valve material]. AB - In this study, nitrogen ion implantation and titanium oxide film synthesized by ion beam enhanced deposition were adopted to treat low temperature isotropic pyrolytic carbon (LTIC). The chemical composition and valence state of the modified layer were determined by X-ray photoelectron spectroscopy (XPS) and Auger eletron spectroscopy (AES). The electric resisitance and the surface energy were measured by the four-probe system and the contact angle method respectively. The blood compatibility of the modified LTIC was evaluated by clotting time measurement and platelet adhesion test. The results showed that carbon-nitride was formed on the LTIC surface after N+ ion implantation, which had some effect on the improvement of blood compatibility, and a significant improvement of blood compatibility of LTIC coated by TiO2-x film was verified. PMID- 12552647 TI - [The biocompatibility of Chinese nitinol alloy intravascular stent]. AB - The aim of this study was to evaluate the biocompatibility of Chinese nitinol alloy intravascular stent. Sixteen Chinese nitinol alloy intravascular stents with 4 mm ID were implanted in the left and right iliac arteries of 8 normal dogs and were observed for one to six months. The results showed that the stents implanted were completely covered by a thin layer of the neointima after one month and the surface of neointima was covered by a monolayer of endothelium-like cells. These suggest that the Chinese nitinol alloy has very good biocompatibility. PMID- 12552648 TI - [Biocompatibility of self-designed absorbable hydroxyapatite/poly (DL-lactide) composites]. AB - The biocompatibility of self-designed hydroxyapatite/poly (DL-lactide) rods was evaluated both in vitro and in vivo including Ames test, micronucleus test, acute and subacute systemic toxicity test, hemolysis test, hemopexis test and long-term muscle and bone implant test. The results indicate that the material has no toxicity, no stimulation and mutation, and it does not cause hemolysis and hemopexis. Consequently, the biocompatibility of the composite was good. PMID- 12552649 TI - [Design of HA/Ti biomedical implants with the use of ion-beam-assisted deposition]. AB - Hydroxy-apatite (Ca10(PO4)6(OH)2,HA) is a potential material for coating on metal implants owing to its bioaffinity and osteo-integration to body. At present, in addition to bulk HA directly used in restoration and reconstruction surgery, hydroxy-apatite is also coated on metal implants by plasma-spraying method, adhesion and sintering. However, these kinds of biomaterials are restricted in application due to their insufficient strength and weak bonding. It is of great advantage to modify the surface of titanium and its alloy by ion-beam-assisted deposition (IBAD) combining ion implantation with physical vapor deposition(PVD). This paper is focused on the design of HA/Ti implants by ion-beam-assisted deposition technique and the computation of fabricating parameter using Trim96 program. The present investigation lays a foundation of further researches and application. PMID- 12552650 TI - [Study on the biologic function of extracorporeal bioartificial liver support system]. AB - In this study, a total of 2 x 10(8) hepatocytes and nonparenchymal liver cells cultured as aggregate spheroids and a hollow fiber bioreactor were used to set up the extracorporeal bioartificial liver support system (EBLSS). Its function was determined throungh the analysis of some cells production. After 6 hours circulating of EBLSS, the total protein, albumin, alpha-fetoprotein and urea were markedly increased in the circulating media, and the DNA synthesis of rat hepatocytes cultured in the media was significantly increased (compared with control group, P < 0.01). This suggests that the EBLSS which consists of cultured human liver cells and hollow fiber bioreactor has the biologic function of liver and has a stimulative action on the proliferation of hepatocytes. PMID- 12552651 TI - [Preparation of collagen-based materials for wound dressing]. AB - This article addressed the methods to develop collagen-based materials for wound dressing. Fresh frozen bovine tendon was treated with 0.05 M acetic acid at pH 3.2 for 48-72 h, then it was homogenized, filtered, and after the process of adding chondroitin sulphate(8%), it was dbubblized; thus 1.5%-2.5% collagen solution was finally prepared. The solution was lyophilized in prefrozen or un prefrozen mold. The collagen sponge was crosslinked with 0.25% glutaraldehyde for 24 h. Three other wound dressings were developed by similar method, namely collagen membrane with polyurethane membrane onlay, polyurethane-coated collagen membrane and collagen membrane on gauze. It was demonstrated that the property of frozen bovine tendon was stable, and the prepared collagen sponge contained pores of 50-400 microns in diameter. Some factors influencing pore structure and size in the collagen sponge were discussed. The results of this preliminary study suggested that the collagen could be used as wound dressing. PMID- 12552652 TI - [Formation and characteristics of an artificial dermis--collagen-chondroitin-6 sulfate membrane]. AB - In this study, we extracted type-I collagen from rat tail tendon with acetic acid and then mixed it with chondroitin-6-sulfate to form a collagen-chondroitin-6 sulfate membrane by freeze drying and crosslinking. The result showed that the collagen-chondroitin-6-sulfate membrane was semitransparent, with a network structure, and the pore size was about 100-150 microns. It was also demonstrated that the "dermis" achieved a good adherence to wound and the degradation rate in vitro was markedly decreased by glutaraldehyde crosslinkage. PMID- 12552653 TI - [A study on adhesive properties of transformed human embryonic tendon cells to PLA and PLGA]. AB - To study the adhesive properties of transformed human embryonic tendon cells (THETCs) to polylactic acid(PLA) and polylactic-co-glycolic acid 85/15 (PLGA 85/15), the adhesive mechanical properties of single THETCs to PLA films (average thickness 18.6 microns) and PLGA 85/15 films (average thickness 17.3 microns) were measured by means of micropipet aspirating technique. The rsults showed that the attachment rate and adhesive force for two polymer groups(PLA precoated with bovine serum albumin (BSA) and PLGA 85/15 precoated with BSA) were significantly lower than those for two control groups(PLA and PLGA 85/15), respectively, while the attachment rate and adhesive force for other two polymer groups (PLA precoated with poly-D-lysine (PDL) and PLGA 85/15 precoated with PDL) were significantly higher than those for two control groups, respectively, and that the adhesive properties of THETCs to PLGA 85/15 films were better than those of THETCs to PLA films. These demonstrate that BSA can inhibit THETCs attachment to polymer, and PDL can promote THETCs attachment to polymer, and suggest that PLGA 85/15 films were better than those of THETCs to PLA films. These demonstrate that BSA can inhibit THETCs attachment to polymer, and PDL can promote THETCs attachment to polymer, and suggest that PLGA 85/15 coated with PDL may be used as a new type attachment substrate for THETCs. PMID- 12552654 TI - [An experimental method for measuing flexor pulley's mechanical effects by imitating hand in vivo]. AB - Flexor pulley system has an important mechanical effect on digital flexion function. An experimental method was designed to measure the pulleys' mechanical effects by using hand in vitro to imitate hands in vivo. The pulleys' function was evaluated by pulley incision and pulley preservation, and percentage loss of work was used as an index. The results demonstrate that this experiment is reasonable in design and methodology. PMID- 12552655 TI - [Changes in rat erythrocyte hemorheology caused by morphine dependence]. AB - This study was aimed at the effect of morphine on the biomechanics properties of rat erythrocyte membrane in vivo and in vitro. The shear elastic modulus (E) and the surface viscotsity viscosity (micron) of rat erythrocyte membrane was measured by using the method of the new ektacytometry. The changes in hemorheology caused by morphine dependence were investigated and the mechanisms of the changes are suggested. PMID- 12552656 TI - [Effects of microwave acute irradiation on biomechanic properties of rabbit tissues]. AB - The rabbit body was irradiated acutely by 2.45 GHz continuous microwave. After irradiation, the values of blood viscosity, blood viscoelasticity, platelet aggregation, erythrocyte fragility, erythrocyte deformation, erythrocyte aggregation, the compressive mechanic property and flexible property of bones were tested. The data of irradiated group and unirradiated group were compared. The results showed: after microwave irradiation, the rabbit blood viscosity and viscoelasticity decreased; the rabbit platelet aggregation rate decreased and the disaggregation rate increased; the erythrocyte fragility increased; the biomechanic properties of bones did not change considerably. PMID- 12552657 TI - [A wavelet neural network algorithm of EEG signals data compression and spikes recognition]. AB - A novel method of EEG signals compression representation and epileptiform spikes recognition based on wavelet neural network and its algorithm is presented. The wavelet network not only can compress data effectively but also can recover original signal. In addition, the characters of the spikes and the spike-slow rhythm are auto-detected from the time-frequency isoline of EEG signal. This method is well worth using in the field of the electrophysiological signal processing and time-frequency analyzing. PMID- 12552658 TI - [Wavelet application to reduction of stimulus artifact in transient evoked otoacoustic emissions testing]. AB - Otoacoustic emissions(OAEs) are presently considered as an objective clinical test for assessing the integraity of peripheral hearing. Owing to the adding of the reflex waves of the stimuli, recordings of emissions evoked in response to transient stimuli are contaminated by an initial artifact, which inhibits the examination of high frequency cochlear responses that have short latencies. So OAEs testing is of serious limitation in most clinical environments. We propose an artifact reduction technique based on discrete wavelet transform pre processing method and demonstrate empirically that the method not only improves artifact reduction but also enhances signal-to-noise ratio in the response region. PMID- 12552659 TI - [The alternation current conductive property of blood]. AB - The blood electro-frequency properties of normal human body were investigated in the range of 1 Hz-10 MHz. The method of testing and the equipment were examined to meet the need for the measurement of blood property. In the experiment, it was discovered that the electric conductivity of blood increased obviously with the increase of testing frequency in the rage of 100 Hz-10 KHz, but almost no change in phase was noted. This result suggests it is necessary to modify the previous work and build up a reasonable equivalent electric model of blood. PMID- 12552660 TI - [Methodological study of measuring flow field uniformity downstream of mitral valve using Doppler echocardiography]. AB - The objective of this study was to establish a quantitative measurement of flow field uniformity of mitral valve in vivo. The flow field uniformity downstream of mitral valve in normal persons and patients with mitral stenosis were studied using advanced color Doppler flow imaging technique. The results showed that a significant difference was found in the quantitative items of flow field uniformity between normal persons and patients with mitral stenosis (P < 0.05). There was significant correlativity between the items of flow field uniformity and other routine items of Doppler echocardiography. The authors conclude that the method is safe, non-invasive, accurate and repeatable, and it can really reflect the flow field uniformity of normal persons and patients with mitral stenosis. PMID- 12552661 TI - [A primary clinical study of HRV analysis of nonlinear dynamics]. AB - We observed the changes of nonlinear dynamic parameters in the patients with diabetes and old myocardial infarction with the methods of nonlinear dynamics. The results indicate that the correlation dimension of the patients with old myocardial infarction is higher than that of the normal, their Lyapunov exponent is lower than the normal's; that the correlation dimension of the patients with diabetes is also higher than that of the normal, their Lyapunov exponent is lower than the normal's; and that the values of the entropy of the two patient groups are basically identical to the entropy of the normal. Therefore the correlation dimension can indicate the changes in the function of autonomic nerve of the patients with diabetes and old myocardial infarction. PMID- 12552662 TI - [The basic study of noninvasive detecting the ischemic/hypoxic cerebral injury]. AB - The cerebrovacular diseases are important causes of mortality and disability in the world, and with the development of science and technology, many mission oriented basic researches are aimed at the ischemic/hypoxic brain injury. This paper introduces some progress made in our research in this field, which includes: the animal experiment model of brain injury; the technique to efficiently extract evoked potentials from electroencephalogram; and the identification of the characteristic information of brain injury. PMID- 12552663 TI - [A computer-aided analysis system for hypertensive retinal image and its clinical application]. AB - With the use of computer image processing technology and microcirculational network analysis technology, we have developed a new system to analyze the outlook of retinal vessles quantitatively. A multimedia structure has been designed for patient's case history file, which includes the basic information of patients, the operating information during sampling and processing patient's retinal image and the result. The system includes image sampling, image displaying, image processing, parameters measuring and analyzing, reports printing and atlas management. Clinical application indicates that the system is useful to hypertension diagnosis. PMID- 12552664 TI - [The prospects of research on VR rehabilitation engineering]. AB - In this paper, we present the recent development of Vitual Reality (VR) technology and rehabilitation engineering and discuss the rationality and feasibility of the application of VR to rehabilitation. With VR technology, psychological induction can be better introduced in rehabilitating exercises and patients can be tremendously released from fret during traditional physical training for rehabilitation. The VR is expected to bring about a technological revolution in rehabilitation engineering. PMID- 12552665 TI - [Study and application on the device of the drop volume of amniotic fluid in estimating fetal lung maturity]. AB - A novel device of drop volume of amniotic fluid has been made to estimate fetal lung maturity. The design of the device is based on the principle that the surface tension of amniotic fluid is inversely proportional to the quantity of surfactant in the solution. The device measures the falling time of fluid drops from drop-tube and estimates the fetal lung maturity by using the method of comparison and electric-optic timing-counting. 148 samples of amniotic fluid were measured by the device and the data were compared with those obtained by biochemical method. The results demonstrate that this device has high stability and accuracy, and the rate of agreement between the two methods is as high as 93%. PMID- 12552666 TI - [A study of time-dependent blood flow distribution in ischemic myocardium and its dependence on perfusion pressure with modeling and emulation]. AB - A mathematical model was proposed for studying the time-dependent blood flow distribution in ischemic myocardium with different developments of collateral circulation. The model includes the blood perfusion of normal myocardium and ischemic myocardium. There are three classical myocardial vascular nets in each part and each net is represented by a branch of time-dependent nonlinear resistance and compliance. Experimentally measured aortic pressure and left ventricular pressure from six male dogs were taken as input data for emulation. The results demonstrate that raising perfusion pressure with external counterpulsation can improve the blood perfusion condition in ischemic myocardium. PMID- 12552667 TI - [Analysis of Parkinson's EEG based on the complexity measure]. AB - EEG represents the electricy activity of neurons in human brain; it is of course repeatedly used for studying and analyzing the brain activity and the status of brain function. We analyzed the Parkinson's and normal persons' EEG, with the aid of Kc complexity and C1 complexity defined by Kolmogorov and Xujinhua respectively. The results of the analysis on the data from 200 patients and 200 normal persons indicate that the complexity of EEG can be regarded as a feature classifying the Parkinson's EEG and normal persons' EEG. PMID- 12552668 TI - [Autodetection of cell adhesion by light transmission]. AB - To set up a new effective method for measuring the adhesion between red blood cells and vascular endothelial cells, we studied the relationship between adhesive red blood cells and light transmission by using Microscope-Photometric Monolayer-Technique Elias-c Analyzer in our department. The results showed that the method of light transmission measurement on the adhesion between red blood cells and vascular endothelial cells is simple, effective and reliable. PMID- 12552669 TI - [Inverse problem identification of parameters in heat transfer processes of human body]. AB - In order that the distortion of the relative skin temperatures which is accompanied with the physiological destruction of an organ in the abdominal cavity and its physical-physiological mechanism may be investigated, we adopt in this paper the mathematical model for heat transfer problems in human layered tissues and a perfect parametric identification approach-inverse problem method. By utilizing the extremum method and integrating with the experimental data of an artificial thermo-focus, this difficult biophysical problem is solved. PMID- 12552670 TI - [Comprehensive evaluation of medical process by constructed polynory function]. AB - This paper describes the concept of "medical process evaluation" and presents a model which is based on constructed polynory function by using the Fuzzy Set theory. This model frame has been used for the prognostic evaluation of patients recovering from acute myocardial infarction and for the comprehensive evaluation of efficacy in treatments of pneumoconiosis. The results are satisfactory. This model frame can be generalized to other medical process evaluation. PMID- 12552671 TI - [Study of methods for measuring spontaneous activation of neutrophils in vitro]. AB - From the viewpoints of cellular morphology and enzymic histochemistry, we adopted the pseudopod method and NBT test for the measurement of spontaneous activation of neutrophils in vitro The time course curves of spontaneous activation of neutrophils from healthy donor were drawn. After comparing the pseudopod method with NBT test, we conclude that pseudopod method is more sensitive and reliable than NBT test and it can be used for real time observation; and that the curves of time course ascend from lower level to the highest level within 3 hours after the samples are collected, and then they descend gradually. The methodology of this study is available for use in future studies on the relationship between activation of neutrophils and many diseases. PMID- 12552672 TI - [A method of preparing the section of eye tissue for transmission electron microscopy]. AB - A method of preparing the section of eye tissue for transmission electron microscopy (TEM) is recommended. The tissue was prefixed with a mixed solution of 4% paraformaldehyde and 2.5% glutaraldehyde and followed by softening in 3% EDTA solution for 20 minutes, then the tissue was fixed in 1% osmium tetroxide, dehydrated in series acetone, infiltrated in Epox 812 for a longer, and embeded. Ultrathin sections were cut with glass knives or diamond knife, stained with uranyl acetate and lead citrate and examined with H600-IV. The advantage of this method is that the ultrastructures of tissues are well preserved without any damage and distortion. The main points of the procedure of preparation have been discussed. PMID- 12552673 TI - [Investigating the phenomenon of total erythrocyte destruction during an in-vitro hemolysis test by roller pump]. AB - This study sought to address the question, "will the phenomenon of 'total erythrocyte destruction' come about during a long time of in vitro hemolysis test by roller pump". One-day-old boving blood was employed in the test by Polystan pump(group A) and Cobe pump (group B). TMB method was used to measure the level of plasma-free hemoglobin. The samples of plasma-free hemoglobin were taken at the time before pumping and pumping for 4,6,8,10, 12,14 and 16 hours. The index of hemolysis was calculated according to Koller's formula. The results showed that the levels of plasma-free hemoglobin in the two groups both increased linearly during the puming of blood. The index of hemolysis was 0.2960 mg/L group A, and 0.3993 mg/L in group B; no statistically significant difference was noted between the two groups. These data indicated that no phenomenon of "total erythrocyte destruction" was observed in this hemolysis test by roller pump. PMID- 12552674 TI - [Progress in studies on cavitation of mechanical heart valve]. AB - This paper reviews the researches on cavitation of mechanical heart valve in recent years and presents a discussion on the research methods. The achievements are summarized and the prospects of further research on cavitation in mechanical heart valve are brought forth. PMID- 12552675 TI - [Differentiation of cultural endothelial cells]. AB - The disparity of function and ultrastructural characteristics between cells in vivo and cultural cells in vitro has been attributed to the differentiation of cells, so differentiation is the key to detecting the differences between endothelial cells (ECs). The ECs are widely distributed over the entire inner surface of blood vessels in the body and exhibit noticeable heterogeneity. The cultural ECs also show heterogeneity. This is a literature review focusing on the recent advance in researches on the embryonic origin of ECs, the heterogeneity of ECs and the factors that influence differentiation of ECs. Also described is the prospect of studies aimed at elucidating the molecular mechanisms involved in the differentiation of ECs. It is hoped that future studies will enhance the realization of oriented differentiation of ECs. PMID- 12552676 TI - [Registration and fusion of multimodality medical images]. AB - This paper introduces the present situation of research and methods of multimodality medical images fusion. Some algorithms and relative techniques are given in detail. A summary of the clinical application of multimodality medical images fusion is also presented. PMID- 12552677 TI - [An improvement in oxygen-drained device for medical hyperbaric oxygen chamber]. AB - This paper introduces a method for improving the oxygen-drained device of hyperbaric oxygen chamber. With the use of the improved device, the density of oxygen in the chamber can be lowered. PMID- 12552678 TI - [Study on the separation of optical chromatography]. AB - The relationship between the particle radius with the intensity of the applied laser radiation force is theoretically studied using a ray-optics model by analyzing the forced-status of the separated particles in the mobile phase. The laser radiation force is proportional to the square of the particle radius when the particle radius is much smaller than the laser beam radius under the condition of using a single-transverse-mode laser beam with a Gaussian intensity distribution and the particle refractive index being larger than that of the mobile phase. The retention distance is calculated and the dynamic range is discussed. Theoretically, the dynamic range of the optical chromatography can be extended by increasing laser power, decreasing the flow rate of mobile phase and focusing the laser beam. PMID- 12552679 TI - [Deductive inference on the plate height equation of electrochromatography]. AB - In this paper, the dynamic process of electrochromatography has been studied. The differences between electrochromatography and high performance liquid chromatography have been compared. The author considers that because of the effects of electroendosmosis flow at the surface and interior of stationary phase particles, the plate height increment due to axial dispersion of the eluite in the interstitial space and the intraparticular diffusion resistance to mass transfer will reduce, and the "film" resistance at the particle boundery will disappear. But when a current passes along the electrochromatographic column Ohmic heat releases and the tube will be heated up. The temperature difference between the center of tube and the tube wall will affect the plate height. The plate height contribution from this is quite significant and can seriously reduce the efficiency of column. Thus, a general plate height equation has been derived to express the effect of axial dispersion in the electrochromatographic process, mass transfer resistances at the mobile phase, kinetic resistances associated with the reversible binding of eluite by the stationary phase and the temperature distribution effect. According to these theories, the plate height equation of electrochromatography has been obtained as following: [formula: see text] It is suggested that there exist a lot of factors which influence the column efficiency of electrochromatography, such as axial dispersion in the interstitial space, mass transfer resistances at the mobile phase, kinetic resistances with the reversible binding of eluite by the stationary phase, and the temperature field in the column inside. The influence of temperature field is closely related with internal diameter of column. PMID- 12552680 TI - [Studies on the simultaneous measurement of several cephalosporins by RP-HPLC (I)]. AB - This paper reported the research on the simultaneous separation and determination of six cephalosporins by RP-HPLC. Six cephalosporins are cefalcor, cefalexin, cefradine, cefadroxil, cefominox and cefoxitin. The analytical conditions for this method were as follows: a Hypersil ODS C18(200 mm x 4.6 mm i.d., 5 microns), detection wavelength: 254 nm; a mobile phase solution of 50 mmol/L monopotassium phosphate (pH 3.4)-acetonitrile (87.5:12.5) and DAD detector. The flow rate was 1.0 mL/min. The calibration curves of the six compounds were linear, the correlation coefficients were 0.9951 for cefominox, 0.9999 for the others, the range were 164 ng-16.4 micrograms for cefominox, 99 ng-9.934 micrograms for cefadroxil, 104 ng-10.358 micrograms for cefalcor, 122 ng-12.224 micrograms for cefalexin, 107 ng-10.702 micrograms for cefradine and 115 ng-11.506 micrograms for cefoxitin. The recovery rates were 103.5% for cefominox, 99.3% for cefadroxil, 101.4% for cefalcor, 101.5% for cefalexin, 98.7% for cefradine and 97.6% for cefoxitin. Six cephalosporins were all stable in 50 mmol/L monopotassium phosphate (pH 3.4-4.6). When preparations of these cephalosporins were determined, it is indicated there were no difference between the results by using this method and the pharmacopoeia methods. The total separation time of these cephalosporins was within fifteen minutes. This method is simple, sensitive, rapid and accurate. PMID- 12552681 TI - [Study on adsorption of atrazine by soil leaching column chromatography]. AB - A recently developed soil leaching column chromatography was applied to study the adsorption of pesticide in water-soil system. Soil was filled directly into a liquid chromatographic column and pesticide solution was applied as an eluant, and the adsorption equilibrium was monitored by a UV detector. The herbicide atrazine, i. e. 2-chloro-4-ethylamino-6-isopropylamino-s-triazine, was chosen as an example. Its mass concentration ranged from 19.73 mg/L to 29.60 g/L. After the soil phase was rinsed with methanol, the amount of atrazine adsorbed was calculated from the difference between the atrazine totally rinsed and that in solution in the column. The low content atrazine sample was determined by enrichment on an ODS column and step-gradient method. It shows that the adsorption of atrazine on a sandy loam soil is in accordance with Freundlich isotherm. The adsorption coefficient Kf is 842.6 mL/kg and the normalized organic carbon adsorption coefficient log Koc is 1.832. PMID- 12552682 TI - [Key factors in the control of electroosmosis with external radial electric field in CE]. AB - Direct control of electroosmosis flow (EOF) by external radial electric field was performed at room temperature using a home-made field-modulated capillary electrophoresis (CE) system. The EOF was monitored at 206 nm by using DMSO as a probe. To apply a radial electric field across the CE capillary wall, the capillary was cased with a wide column. Both of the concentric space and the capillary bore were then filled with an identical running buffer and applied with an axial electric field of 150 V/cm but starting from different levels. All of the tubes used were made of fused silica with polyimide over-coating (from the Yongnian Optical Fiber Work, Hebei, P. R. China). The size of the CE capillaries adopted was 25-100 microns i.d. (375 microns o.d.) x 28.5/45 cm (effective/total length), and that of the casing column 400 microns i.d. x 32 cm. To investigate the fundamentals of the external EOF control when using the flexible fused silica capillaries, various parameters have been inspected such as pH, buffer composition, additives and capillary wall feature etc.. As expected, to well control both of the magnitude and direction of the electroosmosis, the buffer pH should be kept below 4 and the buffer concentration below 50 mmol/L. However, buffers below 1 mmol/L should be avoided because such a diluted running buffer may result in poor CE separation. Weak electrolytes like citric acid, tartaric acid and acetic acid were found to be capable of generating better EOF control than the strong electrolytes such as phosphate and chlorides. This is possibly due to the formation of looser electric double layer with the weak rather than the strong electrolytes. Some wall coatings like calix arene and its derivatives can evidently improve the EOF control even at pH 5. This reveals an exciting way to expend the controllable pH range. In addition, narrow-bore capillaries were demonstrated to be better than wide-bore tubes. Other conditions such as buffer additives and capillary rinse procedure were shown to have only negligible influence on the control. PMID- 12552683 TI - [Preparation and chromatographic characteristics of linear [60] fullerene polysiloxane stationary phase for capillary gas chromatography]. AB - Hydrosilylation of polymethylhydrosiloxane (active hydrogen content was 0.05%) and reactions with allylbromide (or undecenyl bromide), sodium azide and [60] fullerene in their given orders is described. Two kinds of fullerene-containing polysiloxanes were obtained [polymer(3)]. Their [60] fullerene contents were 30 mg (C60-PSO-1) and 60 mg (C60-PSO-2) respectively. These polymers (3) were characterized by FT-IR, 1H-NMR, UV-V, 13C-NMR and used as stationary phase for capillary chromatography. Two fused-silica capillary columns (0.25 mm i.d.) were coated with a 5 g/L solution of polymer (3) in dichloromethane by the static method. The column efficiencies were 3,450 plates/m (C60-PSO-1) and 3,020 plates/m (C60-PSO-2), for naphthalene at 120 degrees C. The average polarities of these columns, represented by the McReynold's Constant, were 79 and 93 respectively. The polarity of C60-PSO-2 was a little higher than that of C60-PSO 1, which was in turn a little higher than that of SE-30(43). These columns showed wide operation temperature range (100-360 degrees C) and outstanding thermostability. It could be used at 360 degrees C with a baseline drift of 30-40 fA. Grob test mixtures eluted on these columns in the ordre of 1,3-butanediol, n decane, 1-octanol, n-undecane, 2,6-dimethylphenol, n-dodecane, 2,4 dimethylaniline, naphthalene, indicating that the strong pi-pi interaction of [60] fullerene with aromatic moieties played an important role in separation mechanism. The columns exhibited unique selectivity for many organic compounds, such as alkanes, alcohols, ketones and anilines. It can well separate benzene homologues, naphthalene homologues and polyaromatic hydrocarbons(PAHs) due to the pi-pi interaction of three-dimensional pi system of C60 and aromatic hydrocabons. And the columns are especially good for the separation of positional isomers(nonanol, butanediol, nitrotoluene, nitrochlorobenzene, nitroethylbenzene, dinitrobenzene, chlorophenol). The columns also have excellent thermostability and are particularly suitable for the separation of high boiling compounds. For example, polycyclic aromatic hydrocarbons and fatty acid methyl esters are well separated on them. PMID- 12552684 TI - [beta-Cyclodextrin bonded stationary phases for chiral separation in HPLC]. AB - A review of beta-cyclodextrin bonded stationary phases in chiral liquid chromatography is given, focusing on design and synthesis of beta-cyclodextrin bonded chiral stationary phases. Fifty-three references are presented in this paper. PMID- 12552686 TI - [Simulation of gas chromatographic peak motion process]. AB - Solute motion process under different temperature, pressure and/or flow program conditions is simulated with the aid of numerical calculation of the differential equation. It is demonstrated that there exist some differences between the normalized average linear velocity and the normalized local linear velocity. As time develops, they intercept, and may not reach the column end at the same time, thus may result in errors in retention time simulation. In constant pressure process, where the compressibility factor (j) does not change, simulation results in a model index of 1, and no model difference can be found when the integrate equation is used. In pressure varying process, where j changes by time, simulation results in an model index which deviates to 1, and positive (for a j decrease process) or negative (for a j increase process) error can be found when the integrate equation is used. The numerical solution of the differential equation is singular under given programs, which corresponds to a specific space line in z, t, u coordinates. PMID- 12552685 TI - [Eluents of non-suppressed ion exchange chromatography with conductometric detection]. AB - A review with 64 references is presented on eluents for non-suppressed ion exchange chromatography with conductometric detection. The kinds of characteristics of eluents in non-suppressed anion exchange chromatography and non-suppressed cation exchange chromatography are described separately. PMID- 12552687 TI - [The compositional analysis of phospholipids and their fatty acids in rabbit sarcoplasmic reticulum]. AB - Phospholipids were extracted from the sarcoplasmic reticulum of rabbit skeletal muscle, and separated by high performance thin layer chromatography. The mole fraction of individual phospholipids were determined by assaying phosphorus of each band. Main phospholipids were scraped and extracted, and subjected to esterification by alkaline methanol, and then the composition and content of fatty acids were determined by gas chromatography. The results show that the main phospholipid components were PC (phosphatidylcholine) and PE (phosphatidylethanolamine). They account for 66.5% and 15.5%, respectively. The main fatty acid in PC is palmitic acid and those in PE are stearic acid and arachidic acid. PMID- 12552688 TI - [Determination of carboxylic acid on a new capillary column prepared by ourselves]. AB - In this paper, use of tri-n-butylammonium dodecylsulfonate as liquid phase to prepare a capillary column suitable to separate carboxylic acids in a direct injection mode, by the method of dynamic coating with mercury plug, is reported. The results showed that the column has good gas chromatographic properties and high selectivity, especially to strong polar compounds, such as carboxylic acids. The preparation of the capillary column, separation and quantitative analysis of carboxylic acids were investigated. Several examples are given. PMID- 12552689 TI - [Quantitative determination of mass ratio of additives in special super-high pressure hydraulic oil]. AB - Two additives, phenol, 2,6-bis(1,1-dimethylethyl)-4-methyl- and triphenyl phosphate with mass ratio of less than one thousandth in special super-high pressure hydraulic oil were determined by GC-MS. Tributyl phosphate ester was used as internal standard. The correction factor of each additive was determined before analysis. Each sample was analysed for 5 times to get good precision. It is satisfactory to use SIM as the detecting mode, and the CVs of correction factors and mass ratio were about 5%. The problems of how to select the monitoring ion in SIM mode and the sample size required are discussed in this paper. This method is satisfactory in analyzing low mass ratio constituents in a mixture. PMID- 12552690 TI - [Gas chromatographic and gas chromatography-mass spectrometric analyses of aromatic hydrocarbons in oils and rock extracts simultaneously by using bench-top gas chromatograph-mass spectrometer]. AB - This article reports a successful example that an injected sample is simultaneously introduced into two fused silica capillary columns and detected separately by a flame ionization detector (FID) and an HP 5973 mass selective detector (MSD) both installed on an HP 6890 Gas Chromatograph. This GC and GC/MS analyses of aromatic hydrocarbons in oils and rock extracts were carried out by using this method and good results were achieved. This method is of great value in organic geochemistry research because it is efficient and economical. PMID- 12552691 TI - [Determination of phospholipids in bear bile by isocratic high performance liquid chromatography]. AB - A high performance liquid chromatographic method for the determination of phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol (PI) in the bear bile was established. The air-dried bile of bear was ground into powder and extracted continuously with V(chloroform):V(methanol) = 1:1 to a final dilution 25-fold the volume of the sample. The extract was washed by addition to it of 0.5 its volume of 0.58% aqueous NaCl. The resulting mixture separated into two phases. The lower phase was the total pure lipid extract and was diluted to the desired volume by addition of V(chloroform):V(methanol) = 1:1 mixture and then injected into the HPLC system. The conditions of HPLC were P-E silica column (150 mm x 4.6 mm i.d., 10 microns); V(acetonitrile):V(methanol) = 76:24 mobile phase with a flow rate of 1.5 mL/min and UV detector at 205 nm. The linear regression equation was Y = 5,917X + 1,709(r = 0.9970) for PI, Y = 7,362.1X 6,608.2(r = 0.9949) for PE and Y = 2,457X-2,460.5(r = 0.9972) for PC. The linear range were 1.0-12.0 micrograms, 2.0-24.0 micrograms and 9.5-114.0 micrograms respectively, The relative standard deviation of peaks were 1.6%, 0.89% and 2.6% respectively. The average recovery of PC was 89.30% (RSD = 2.0%). The method enables a simple, rapid and reproducible quantification of PI, PE and PC. PMID- 12552692 TI - [Immunoaffinity purification of specific immunoglobulin from egg yolk]. AB - Immunoaffinity column (Sepharose-4B) was made with bovine serum albumin (BSA) and used to isolate anti-BSA antibody from egg laid by hens which were immunized with BSA. Polyclonal antibody was eluted under different conditions (pH 5.0-2.8) because of its different affinity against antigen. SDS-PAGE and double immunodiffusion analysis confirmed that antibody isolated from egg yolk was electrophoretically pure and specific. According to the separation aim in this paper, the final elution buffer was 0.1 mol/L glycine-HCl buffer, pH 2.8. The final antibody yield was higher than 90%. As a new development in chromatographic media, POROS has its maximum pressure limit of 21 MPa. It has been widely used because of its high performance, high flow and large capacity. The sugar residue of the antibody was then oxidized and coupled to the hydrazide activated POROS HY. Pure targeted protein (BSA) was obtained through the POROS HY column. The tendency of specific antibody production was investigated during the immunization period. The amount of specific antibody has increased obviously after boost immunization. PMID- 12552693 TI - [A novel chiral selector in capillary electrophoresis--beta-cyclodextrin polymer]. AB - Three chiral selectors CM-beta-CD, EP-beta-CD, beta-CD were studied for the enatio-separation of three drugs under optimum conditions respectively. The results demonstrate that the resolving power for the drugs is as follows CM-beta CD > EP-beta-CD > beta-CD, with the exception of lobeline. This is due to the- CH2COOCH3 group of CM-beta-CD, which will change the combination and improve the recognition on guest molecules. Although EP-beta-CD is inferior to CM-beta-CD for the separation of chlorpheniramine and verapamil, it has excellent recognition on lobeline and it has not been reported previously. In most cases EP-beta-CD is superior to beta-CD. The explanations are: (1) EP-beta-CD has good solubility in water, which enables high concentrations to be used and consequently achieves excellant separation of racemic compounds, (2) the polymerization of beta-CD changes the properties of CD units and the process produces a more rigid and different conformation from CD, (3) we must attribute much merits to the cooperation or synergism of two, three or even more CD moieties of two polymers for inclusion. Complexation with analytes possesses more than one guest part in their structure. PMID- 12552694 TI - [Purity analysis of cephalosporins with capillary zone electrophoresis]. AB - A capillary zone electrophoresis method is proposed for the purity determination of nine cephalosporin drugs. A background electrolyte comprising of either 20 mmol/L pH 9.20 borate buffer or 20 mmol/L pH 6.86 phosphate buffer was used for most drugs studied except for cefaloridine which formed neutral molecules at these pH values. For it 50 mmol/L pH 2.05 phosphate buffer was used instead. Internal normalization method was employed for quantitation. The method is simple, rapid and versatile. Analysis was completed within 8 min. The merits and limitations of the method were also discussed. PMID- 12552695 TI - [Determination of baicalin and chlorogenic acid in yinhuang granule by capillary electrophoresis]. AB - Yinhuang Granule is a kind of complex formulae made up of extract of Flos lonicerae and that of Radix scutellariae, of the fuctions of dispelling wind-heat to reduce fever, detoxicating, relieving inflammation, promoting urination and tranquilizimg the mind and can be applied to upper respiratory tract infection and tonsillitis. Baicalin and chlorogenic acid are referred to main effective components of it. In this paper, the separation and determination of baicalin and chlorogenic acid in Yinhuang Granule by capillary electrophoresis is described. The experimental conditions are stated as follow: separation column: bare fused sillica capillary (50 microns i.d., 370 microns o.d., total length 47 cm, 40 cm from the inlet to UV detector); electrolyte: 25 mmol/L borax, pH 8.5; detection wavelength: 310 nm; sampling: 17 kPa.s; running voltage: 25 kV, polarity from anode (inlet) to cathode (outlelt); temperature: 25 degrees C; internal standard: p-nitrobenzoic acid. The linear ranges of determination for baicalin and chlorogenic acid were 160-960 mg/L(r = 0.9993, RSD = 1.76%-2.33%) and 80-960 mg/L(r = 0.9989, RSD = 1.07%-2.51%), respectively. The recoveries of baicalin and chlorogenic acid were (102.09 + 1.74)% (RSD = 1.71%, n = 6) and (99.81 + 3.11)% (RSD = 3.12%, n = 6). The method is simple, fast and accurate. It can be used for the quality control of Yinhuang Granule and its related Chinese medicines. PMID- 12552696 TI - [Analysis of terpene/maleic anhydride adduct and its derivatives by GC]. AB - The retention behaviors of terpene/maleic anhydride adduct(TMA, 1) and its esterification derivatres(2) on OV-17 and SE-30 columns has been studied. The relationships between adjusted retention time and column temperature for the two compounds were linear. The shape of both peaks was more symmetrical on OV-17 column than on SE-30 column. It was found that the peak area was unaffected when the temperature of injector was higher than 260 degrees C. PMID- 12552697 TI - [Determination of alkaloids in pericarpium papaveris by gas chromatography with wide bore capillary column]. AB - A method for the simultaneous determination of codeine, morphine, thebaine, papaverine and narcotine in pericarpium papaveris by gas chromatography with wide bore capillary column is described. The major components in pericarpium papaveris were extracted with methanol and chloroform by supersonic extraction and evaporation. The residue was dissolved in methanol. The resulting solution was used for analysis. The conditions for determination were: FID detector, HP-1 capillary column, 5 m x 0.53 mm x 2.65 microns film, column temperature 260 degrees C, The method requires 10 minutes for the whole analysis. The average recoveries of codeine, morphine, thebaine, papaverine and narcotine in the sample were 94.0%, 96.5%, 93.8%, 91.0% and 91.4% respectively. The relative standard deviations were from 0.92% to 2.75%. The advantages of this method are simple, rapid, accurate and sensitive. PMID- 12552698 TI - [Single-pump on-line addition of modifier for supercritical fluid chromatography]. AB - Modified CO2 as mobile phase is usually necessary for packed-column supercritical fluid chromatography. Single syringe pump was applied in this work to add modifier through a parallel connection device, which can maintain constant and stable volume fraction of modifier without any contamination of the pump. Viscosity of the modifiers (here are methanol and acetone) and the length of the CO2 pipe line/modifier pipe line can affect the volume fraction. Volume fractions of methanol and acetone were determined by gas chromatography and ultraviolet spectrometry respectively and their rules of variation were also examined. It shows that the volume fraction of methanol is lower than that of acetone under similar condition. With fixed modifierr line, the longer the CO2 line is, the higher the volume fraction will be. The new device can conveniently alter the nature and ratio of the modifier with quite stable volume fraction. The online addition device can tentatively replace the dual-pump system. PMID- 12552699 TI - [Determination of total sulfite in wine by high performance liquid chromatography(HPLC) with fluorescence detection]. AB - A highly sensitive fluorometric method for the determination of total sulfite in wine is reported. The strong fluorescent derivatives formed by the quantitative reaction of sulfite in wine and N-(9-acridinyl) maleimide(NAM) in aqueous solution under mild conditions was injected into the HPLC. Separation was performed on an ODS column(200 mm x 5 mm) with a mobile phase of V(CH3CN):V(0.1 mo/L NH4AC) = 25:75 at a flow rate of 1.0 mL/min and detected by fluorometry at 455 nm(excitation at 365 nm). There was a good linear calibration curve in the range from 0.25-4 mg/L(r = 0.9998), The limit of detection was 1 ng. Sulfite mass concentration in wine has been determined by this method with good results and recoveries. This method is simple, rapid and reliable, and a sample size of only 1 mL is required to complete an analysis. PMID- 12552700 TI - [Determination of acemetacin and indometacin in human serum by high performance liquid chromatography]. AB - A method of HPLC for the quantitative determination of acemetacin and indometacin in human serum is described. After being extracted with diethyl ether, acemetacin and indometacin were analyzed by reversed-phase HPLC(Spherisorb-C8) and UV detector(254 nm), with tolbutamide as internal standard. The mobile phase was a mixture of V(acetate buffer solution, pH 4.6):V(methyl alcohol):V(acetonitrile) = 55:5:40 and at a rate of 1.0 mL/min. Over the mass concentration range of 12.5 micrograms/L-1.6 mg/L, both the calibration curves were linear, r = 0.9996, n = 8. The average recoveries of acemetacin and indometacin were 77.2% and 86.7% respectively. The within-day and between-day RSD of acemetacin and indometacin were less than 5% and 10% respectively. PMID- 12552701 TI - [Detection of residual antibiotics in honey by capillary high performance liquid chromatography]. AB - The quantitative analysis of the three antibiotics-tetracycline(TC), oxytetracycline(OTC) and chlortetracyclinum(CTC) was carried out with a self packed Hypersil C18 150 mm x 0.5 mm i.d. column. As little as 0.5 nanogram of antibiotics can be well detected, the capillary high performance liquid chromatographic method is 100 times more sensitive than that with ordinary HPLC method. The coefficients of linear correlation of mass concentration from 0.5 nanogram to 20 nanogram were rOTC = 0.99695, rTC = 0.99778 and rOTC = 0.98836. It showed that capillary HPLC is a good method used for sensitive analysis of antibiotics in honey. PMID- 12552702 TI - [High performance thin layer chromatographic (HPTLC) determination of PAF in cerebrospinal fluid of patients with subarachnoid hemorrhage(SAH)]. AB - The Platelet Activating Factor (PAF) is believed to be the major function in human after cerebral vascular spasm and cerebral ischemia. PAF has been found to participate in cerebral vascular spasm and cerebral ischemia by the basic and clinical study. The symptom of cerebral vascular spasm and cerebral ischemia has appeared with SAH. It has not been reported that the rule and change of PAF with SAH. In the present work, the concentration of PAF in human cerebral spinal fluid (CSF) with SAH were determined by high performance layer thin chromatography. The TLC plate was coated with high performance silica gel G using V(chloroform):V(methanol):V(water) = 65:35:6 as developing solvent. The PAF was determined by TLC scanning method and detected at 630 nm. The method was applied to determine the concentration of PAF in 16 CSF samples with SAH. The samples were collected in 1-3 d, 7-10 d and 14-21 d. CSF samples were deproteinized with methanol and chloroform. After centrifugation, the chloroform layer separated was dried at room temperature with nitrogen and stored under 20 degrees C in the refrigerator. The linear range of the method was 0.5-2.5 micrograms/L with regression coefficient of 0.9990. The lower limit of detection were 50 ng/L. The recovery of the method was 98.6%. The method enables a simple, rapid and reproducible quantification of PAF with SAH. PMID- 12552703 TI - [Determination of serum proteins by high performance capillary zone electrophoresis]. AB - The separation method of serum proteins was established with an untreared 50 microns i.d. x 47 cm (40 cm to detector) capillary and detection of absorbance at 200 nm. Analysis was performed by pressure injectction 17.23 kPa.s and by applying 23 kV in the constant voltage mode. Serum samples were diluted 40-folds with assay buffer (12.5 mmol/L sodium borate, 1 mmol/L calcium lactate, 0.7 mmol/L magnesium sulfate, 1 mmol/L EDTA were mixed). A normal control serum protein was separated into 6 fractions. In pregnant serum, the alpha 0 was an additionally unknown fraction. Comparison of capillary electrophoresis with conventional cellulose acetate electrophoresis for analysis of serum proteins from normal control, pregnant women multiple myeloma and tonic rachitis patients indicates that capillary clectrophoresis is a new technique for the analysis of serum proteins because of its high efficiency, on-line data processing and automation. Capillary electrophoresis is the reliable technique for clinical diagnosis of serum protein abnormalities. PMID- 12552704 TI - [Determination of 2,5-ditertiary butylhydroquinone by gas chromatography]. AB - Antioxidant 2,5-ditertiary butylhydroquinone was prepared from hydroquinone and tertiary butyl alcohol. A normalized gas chromatographic method for 2,5 ditertiary butylhydroquinone by using an OV-101 fused silica capillary column and flame ionization detector is reported. The quantities of hydroquinone, tertiary butylbenzoquinone, tertiary butylhydroquinone and 2,5-ditertiary butylbenzoquinone mixed with 2,5-ditertiary butylhydroquinone were also determined. PMID- 12552705 TI - [A gas chromatographic method to decrease the detection limit of trace benzene in CS2 extract]. AB - A method to decrease the detection limit of trace benzene in CS2 extract from coastal water by gas chromatography has been studied. A direct injection port (Shimadzu WBI-17) and a 2 m x 2 mm i.d. column packed with Chromosorb W(AW-DMCS) coated with 10% SE-30 was used. It is simpler and has low detection limit, small sample amount and high repeatability. The experiment showed that the trace water in the organic phase and the too small purge flow of the direct injection port could cause serious tailing of CS2 peak. There was an optimum value of the purge flow (purge flow/total flow = 5%). The minimum detectable limit of benzene was 4 micrograms/L. The repeatability (RSD) was better than 6% and the average recovery was 96.7%. PMID- 12552706 TI - [Determination of bentazon and 2,4-D butyl in mixture formulation by HPLC]. AB - Bentazon and 2,4-D butyl(the active ingredients of Cao Di) were determined by reversed phase HPLC aimultancously using methanol/acetonitrile/citrate buffer(pH 3.1) as a mobile phase on Shim-pack clc-CN column. Results showed that the linear correlation and recovery of 2,4-D butyl were 1.000 and (100.2 +/- 0.8)% respectively, and those of bentazon were 1.000 and (98.6 +/- 0.9)% respectively. The coefficients of variation for bentazon and 2,4-D butyl were all was 0.37%. PMID- 12552707 TI - [Analysis of organic additives in copper-plating brightener by high performance liquid chromatography]. AB - This paper describes a high performance liquid chromatographic method for analysis of organic additives in copper-plating brightener. The copper-plating brightener was filtered through 0.45 microgram micropore membrane and determined on a Spherisorb-C6H5 column, with V(methanol):V(water) = 30:70 mobile phase at the flow rate of 1.5 mL/min, and detected at UV 210 nm. 2-benzimidazolethiol, ethylene thiourea, 1-propanesulfonic acid 3,3'-dithio-bis-disodium salt were separated and determined. The recoveries were more than 99% with CV less than 1%. The method is simple and rapid. PMID- 12552708 TI - [Separation and determination of 1,5-benzothiazepines-alpha-chloro-beta-lactam with thin-layer chromatography]. AB - A method for separation and determination of 1,5-benzothiazepines-alpha-chloro beta-lactam in reaction mixture by thin-layer chromatography has been established. The operating conditions were: lambda 1 = 270 nm, lambda 2 = 310 nm, beam size: 1.2 mm x 1.2 mm, SX = 3. The calibration curve was linear in the range of 0.285-5.7 micrograms (r = 0.9952, n = 8) and the detectable limit was 0.114 microgram. The coefficient of variation of this method was less than 5% and the average recovery for beta-lactam was 96.75%. The method is simple and rapid. PMID- 12552709 TI - [Determination of beta-carotene in tobacco by HPTLC]. AB - This paper reports the high performance thin-layer chromatography (HPTLC) determination of beta-carotene in tobacco. The developer used was V(benzene):V(acetone) = 4:1. The detection wavelength was 450 nm. The linear range and the detectable limit of the method were 50 ng-0.5 microgram and 20 ng per spot of beta-carotene respectively. The method has been proved to be simple, sensitive, accurate and reproducible. PMID- 12552710 TI - [Changes of F-actin in neutrophils under fluid shear stress]. AB - Neutrophils are the main defender in the body, and their physiological characters are associated with functions. Because the environments of neutrophils maturation and storage are different from their function environment, they have to adapt thess changes. Filamentous actin (F-actin), as one of the important cytoskeleton components of neutrophils, has different quantity and distribution under different physiological conditions. Previous researches are all focused on static activation of neutrophils using various stimulants such as formyl-methionyl leucyl-phenylalanine (f-MLP) and tumor necrosis factor(TNF). In our study, Low Shear 30 and NXE-1 rheometers were used to provide steady or sinusoidaloscillatory fluid shear stress on separated neutrophils. Fluorescent agent Tritc-Phalloidin was added to label F-actin and the mean fluorescent intensity tested by flow cytometry was used as the indication of F-actin quantity and confocal laser scan microscope was used to detect the distribution of F-actin in neutrophils. We found that F-actin polymerization was significantly decreased under both steady and sinusoidal-oscillatory shear stress when the shear stress was below certain level. The density of the cortex F-actin near cell membrane in cells sheared became thinner than in that without shear stress. f-MLP and TNF both could increase actin polymerization in neutophils due to activation. When neutorphils were activated with f-MLP or TNF under shear stress, the quantity of F-actin in the cells was also decreased, but it was still higher than that in the cells absent of f-MLP or TNF. We concluded that neutrophils had different mechanical properties in different fluid environments. F-actin content and distribution would change according to the shear stress values to play their determinant role in neutrophils mechanical adaptation. The mechanism of actin polymerization in neutrophils for mechanical action is probably different from that for chemoattractants and cytokine activation. PMID- 12552711 TI - [Study of bacterial adhesion to prosthetic valve materials in vitro]. AB - This article reports a study of the bacterial adhesion to prosthetic valve materials in vitro. The method for assessing the degree of bacterial adhesion to prosthetic valve materials was established primarily. The capacities of staphylococcus aureus(SA), staphylococcus epidermidis(SE), Escherichia Coli(EC) and Pseudomonas aeruginosa(PA) for adhesion to Dacron, Pyrolytic carbon and polytetrofluoroethylene (PTFE) were quantitatively determined by the plate counting and Gamma-ray counting of 125I radiolabeled bacteria in vitro. The results showed that the capacities of four types of bacteria for adhesion to Dacron, Pyrolytic carbon and PTFE coincided with the bacterial-growth curve. The capacities of four bacteria for adhering to Dacron were stronger. The adhesion of SE to Pyrolytic carbon was the strongest. The adhesion of PA Kept up a high level. The capacities of EC and PA for adhering to PTFE were the strongest. The results indicate that the capacities of different types of bacteria for adhesion to the same prosthetic valve material are different, and the capacities of one type of bacteria for adhesion to different materials are also different. PMID- 12552712 TI - [Modification of the surface of prosthetic valve material by LGDP for protection against microbial adhesion]. AB - This study aimed at the surface modification of prosthetic valve materials by means of cold glow discharge plasma. Orthogonal design was adopted to select the optimal scheme by the experiments of the singular factor, such as output power, gas flow rate, treatment time etc. The PEG-200 was introducted to the surface. Grafting a layer of heparin onto the surface; coating a layer of PEG-like onto the surface first, and then the grafting heparin. Comparing the adhesive amounts of staphylococcus epidermidis (SE), the adhesive curves for Dacron, which was treated by the above methods; the last one is the best. PMID- 12552713 TI - [Blood-purification characteristics of immobilized L-asparaginase by mathematical model]. AB - Plasma was purified by immoblized L-asparaginase column. The predicted results were in good agreement with the experimental data, which indicated that the mathematical model was suitable for the mass transfer of blood purification. PMID- 12552714 TI - [An experimental study on the change of adhesion between leukocyte and endothelium in acute cerebral ischemia]. AB - The objective of this experiment was to investigate the change of leukocyte adhesion in the focus of cerebral ischemia-reperfusion injury. Immune fluorescent microscope equipped with image tube camera and high speed recording TV sets was used to study the leukocyte adhesion in the focus of cerebral ischemia reperfusion injury. The result showed that the density index of adherent leukocyte was significantly elevated, the fracture stress between leukocyte and endothelium dropped, and the adhesion increased in the focus of cerebral ischmic reperfusion injury. These findings indicated that leukocyte adhesion was increased significantly and leukocytes played an important role in the cerebral ischemia-reperfusion injury. PMID- 12552715 TI - [Studies on degradation of poly-(propyl,3-hydroxypropyl)-DL-asparamide and release of conjugate linking aspirin in vivo and in vitro]. AB - Two polyasparamide derivants having different hydrophilicities were selected to be degraded in vivo and in vitro. The results demonstrate that the degradation process of polyasparamide is enzymatic hydrolysis. Materials with different hydrophobicities are different in degradation rate and the same material has different metabolic rates in the planting position, kidney and liver. The release results of two polymeric drugs indicate that protease is of advantage to release of drug, and hydrophobicity of material is also an advantage to the release. PMID- 12552716 TI - [A water-soluble synthetic polymer, alpha,beta-poly(hydroxyalkyl)-DL-asparamide, and conjugating drug]. AB - A water-soluble polymer alpha,beta-poly (hydroxyalkyl)-DL-asparamide was synthesized by polysuccinimide(PSI) and different lengths of hydroxyalkyls, including alpha,beta-poly(hydroxyethyl)-DL-asparamide(PHEA), alpha,beta poly(hydroxypropyl)-DL-asparamide(PHPA), and alpha,beta-poly(hydroxybutyl)-DL asparamide(PHBA). These polymers were characterized by differential scanning calorimetry(DSC) and infrared spectrophotometry(IR). Stability and acutetoxicity of these polymers were studied. The experiment indicated that these materials were of low-toxicity and high stability. Acetylsalicylic acid, as a model drug, was conjugated into polymers; the drug loadings were 38.63%, 37.68% and 38.70% respectively. Polymer drugs were made into cylinder, and in-vivo release in rabbits was set out. It showed that the longer the spacer was linked into the polymer, the faster the drug was released. PMID- 12552717 TI - [The effect of NaOH on erythrocyte membrane protein band 3]. AB - We used the C1 (Ala 893-Val 911) and KS-2(Ser 731-Lys 743) peptides of band 3 protein as indicators to study the effect of NaOH on the band 3. Five peptides were released from the band 3 molecule when 100 mM NaOH-stripped membrane was digested with trypsin. All of the five peptides were sequenced. PMID- 12552718 TI - [A comparative study of three-dimensional movements of lumbar spine in the old and the young people]. AB - This article is aimed at the differences in lumbar vertebral three-dimensional movement and in stability between the old and the young people, and at the reasons which lead to the differences. Fifteen fresh adult male cadavers were divided into two groups: the young group, 20-30 years old, comprising 7 cases; the old group, 60-70 years old, 8 caes. Lumbar spine(L1-S1) was cut down. Three dimensional movements under a loading system were measured by a compute system. The result showed that the range of the motion of the old group was lower than that of the young group. It suggests that the lumbar spine of the old people is more stable than that of the young people. This may be associated with the degeneration of the lumbar vertebra, which arouses biocompensatory changes, so the lumbar spine becomes re-stable. PMID- 12552719 TI - [Simulation on the hydrodynamics of a flow chamber system]. AB - The distributions of the velocity and the shear stress of flow chamber are achieved by using the nondimensional technique and the method of Bessel function of imaginary argument according to Navier-Stokes' equations. When reduced to the representation of steady flows, the formulae obtained in this paper are consistent with Cao's theoretical formulae and their experimental results. PMID- 12552720 TI - [Computer-aided legal medical examination of body surface]. AB - This paper provides a package of automatic and semi-automatic methods for computing the area of different kinds of body surface injuries. Compared with traditional methods, these processes of examination are faster and the conclusions are more precise and objective. Also presented are the authors classify the items into many types by standards which are necessary to let computer draw conclusions automatically. This software is conducive to improvement in work efficiency and convenience for judicial supervision. PMID- 12552721 TI - [Human motion analysis system based on common video-camera]. AB - A new human motion analysis system based on common video-camera is introduced in this paper. The image processing technique is used to recognize and track anatomic landmarks, which makes data processing more quickly and accurately. The system is cost-effective and can be easily operated because the common video cameras instead of special instruments are used. Furthermore, the system can also be used for detection and analysis of other low-speed moving objects. PMID- 12552722 TI - [Some basic techniques of virtual reality body healthiness vehicles for rehabilitation]. AB - Some know-how techniques of virtual reality rehabilitation vehicles are discussed in this paper after a brief introduction of the application background and the structure and functions of the vehicles. This kind of vehicles can be used for psychological treatment during rehabilitation. Pateints can select different modes of physical exercises according to their individual situations, and therefore they can get rehabilitation much faster and more easily than those under the traditional treatments. PMID- 12552723 TI - [Best minimization to locate the brain sources of magnetic waves]. AB - Locating the sources of brain magnetic fields is a basic question in magnetoencephalography (MEG). This question belongs to the indeterminate problem, that is, infinite solutions can be obtained from the equations of the measured small magnetic fields on the detector. Therefore a model of mathematics is needed to give more information. The major models of mathematics are the search method with many dipoles and the Lp minimum norm method. For the first model, if a global search is used, too much calculation time will be spent; if the gradient search is used, the searches usually converge to a local minimun. So other methods are introduced to find out the initial values, and then the gradient search can be used. For the second model, the depth of the brain sources will dependend on the choice of the weight functions which have several different types, and we do not know in advance which type is correct, so that it is difficult to have a good estimation of the depth of the brain sources. To overcome the difficulties, we use the method with three object functions which are the minimum Lp norm, the deviation between the calculated magnetic fields and the measured magnetic fields, and the number of the sources. This method is similar to the minimum norm method, a weight function is also introduced. But this weight function contains many reference variables which have to be determined through the minimization with three object functions. By choosing suitable weight function and its reference variables, the correct estimation of the depth of the brain sources can be achieved. PMID- 12552724 TI - [A study on distribution modeling of the electrical stimulation under surface nerve stimulation electrodes for somatosensory evoked potential]. AB - The detection of somatosensory evoked potential (SEP) has been an important approach to determining the function of spinal cord and peripheral nerve. Transepidermal Electrical Neuromuscular Stimulation (TENS) on the posterior tibial nerve for SEP is one of the applications and can improve the effectiveness. This paper theoretically studies the distribution of potential field and current density field for human tissue under TENS. A mirror method is used and the analytical solution of the potential distribution is obtained. PMID- 12552725 TI - [Electroporation and its mechanism due to low amplitude transient electromagnetic pulsess on cell membranes]. AB - Low amplitude transient electromagnetic pulses (TEMP) were used to radiate some cells in the broad band transverse EM-wave cell (BTEM Cell). It has been shown that electroporations have been formed on the cell membranes under weak electromagnetic irradiation. The mechanism of electroporation has been analysed preliminarily. PMID- 12552726 TI - [Effects of human thorax tissues on conduction of electrocardiogram and body surface potential]. AB - In this paper, the forward problem of electrocardiogram(ECG) has been calculated by means of numerical simulating techniques. Firstly, It is focused on the effects of organs and tissues within the thorax on the body surface potentials under a 2-D cross section of thorax in terms of finite element method (FEM). To check the effect of skeletal muscle layers on ECG, the simulating computation of ECG, by a method combining FEM with boundary element method (BEM), has been carried out in terms of a 3-D thorax model. PMID- 12552727 TI - [High order multi-layer neural networks and EEG diagnosing]. AB - In this experiment, high-order multi-layer neural networks was introduced into the identification of EEG signal. The result of testing demonstrated that the identification rate reached an a satisfying level. PMID- 12552728 TI - [Neural network approach to medical grading of rheumatoid arthritis]. AB - A grading system based on neural network has been developed and used int he medical grading diagnosis of a case group of Rheumatoid Arthritis presenting Cold syndrome intermingled with Heat syndrome. The possibility of using neural network in TCM diagnosis is preliminarily explored in this paper. PMID- 12552729 TI - [Experimental study of ablation effect on canine esophagus irradiated by pulsed Ho:YAG laser]. AB - To evaluate the influence of power, irradiation time, energy of per pulse and frequency on ablation effect of pulsed Ho:YAG laser, five canine esophagi were studied. We irradiated the mucosa in different power and different irradiation time. For the same power, the samples were divided into two groups: the high energy per pulse low frequency (H)group and the low energy per pulse high frequency (L)group. The two groups were paired. We compared the effects of ablation effect in different laser settings. The results reveal that the ablation effect is positively correlated to the power and irradiation time. The ablation in H group is more obviouse than that in L group. When Ho:YAG is used in high power, ablation is the main biological effect. PMID- 12552730 TI - [The use of inexact reasoning method for differential diagnosis of pulmonary function]. AB - This article expounds the main idea of the evidence theory and hence presents a mathematical model. Based on the above work, we have developped a computer-aided medical differential diagnosis system with Visual Basic. The system can identify restrictive ventilatory impairment, obstructive ventilatory impairment, mixed ventilatory impairment and normal. according to the valuse of patients' related indexed. In a retrospective test of 290 cases we collected, the rate of agreement between the diagnosis made by this system and the physicians was 93.1%. PMID- 12552731 TI - [A comparative study of finger plethysmography and aortic pressure for monitoring the effect of external counterpulsation]. AB - Invasive pressure detecting and monitoring of aortic root was conducted in 20 volunteers so as to study the effect of Enhanced External Counterpulsation (EECP) on aortic pressure and finger plethysmography. The results demonstrated that the changes of aortic diastolic pressure linearly related to the counterpusation's bladder pressure, and the aortic augmentation increased to the top of 168.56 mmHg with a D/S ratio of 2.75(amplitude ratio of diastolic waveform and systolic waveform, D/S ratio) following the bladder pressure of 0.45 Kg/cm2. With the increase of bladder pressure, the dp/dt of aortic augmentation elevated gradually, but a down-regulating trend was seen in systolic waveform. During EECP, the starting time of plethysmographic augmentation had a mean delay of 108.20 +/- 25.70 ms comparing with that of aortic augmentation. The duration between the starting time of aortic systolic waveform and that of finger plethysmogram was arround 96.69 +/- 38.70 ms. When the bladder pressure changed, no significant difference was observed among them. The D/S ratio of finger plethysmographic waveform changed in concord with that of aortic pressure with the promotion of bladder pressure. Therefore, a relationship between aortic D/S ratio (Y) and plethysmographic D/S ratio (x) was constructed by the linear regression analysis: Y = 0.376 + 0.457X. It can be concluded that EECP has significant effects on aortic D/S ratio, aortic peak augmentation and its dp/dt value. Finger plethysmography, to date, still plays important roles in monitoring and controlling EECP work safely and effectively. PMID- 12552732 TI - [A review on deformable model in medical image analysis]. AB - This paper reviews the deformable model in medical image analysis and its applications in registration, motion tracking, segmenting and shape modeling. PMID- 12552733 TI - [Stress-adaptation of bone]. AB - Loading mechanotransduction sensors on the membrane of bone cells or osteoblasts can improve bone mechanisms and influence second messages, growth factors, bone matrix and oncogenes. The response of bone to stree is preportion to the magmititude of the stress. Stress-adaptation of bone is related with the magnititude, perporties, frequency time of stress. PMID- 12552734 TI - [Perspectives of effect on new electrode technology with implantable motor prostheses for stimulating peripheral nerves]. AB - This paper explores approaches to electrode technology and stimulating techniques that selectively activate specific portions of a peripheral nerve trunk with much less implanted hardware than is required with present motor prosthesis systems. The first section reviews the vulnerability of nerves to both mechanical and electrical factors and suggests ideas on how these findings might be utilized in future electrodes. The second section presents results on longitudinally oriented intrafascicular electrodes for nerve stimulation recording. The third section reviews results on longitudinal intrafascicular electrodes for nerve stimulation and recording. PMID- 12552735 TI - [Applications of porous polymeric materials and its biocompatibility]. AB - With the barrier materials, guided tissue regeneration materials and hybrid artificial organs as the representafive materials, the applications of polymer skeletal materials in biomedical fields are introduced in this paper. Based on the results of the induced carcinoma, collagenous encapsulation and chronic inflammation around implanted polymers, the effects of polymers topography on biocompatibility are discussed, and the importance of topographical compatibility of materials is emphasized. PMID- 12552736 TI - [Prospects for antisense techniques]. AB - Antisense technique, developed on the principle of base complementation, is a method using short strand oligonucleotide fragments complementary to target DNAs or target RANs for inhibition of gene expression. As a theoretically perfect and specific method to block the expression of gene, it has been rapidly developed over the last twenty years. This review expatiates on its techniques, its problem and possible way of problem-solving, and its recent application in medicine. Also this review discusses the trends of its further development. PMID- 12552737 TI - [Experimental study on hemoperfusion to remove CIC in blood of rabbit with an affinity adsorbent II]. AB - In this study a rabbitmodel of high concentration cycle immune complex (CIC) was established, and then an experimental hemoperfusion using AA3 Adsor-bent was performed the effect of hemoperfusion to remove CIC in blood and the influence of AA3 on hemocompatibility and proteins were observed. PMID- 12552738 TI - [Technical ensurement of fire-proof in hyperbaric oxyger chambers]. AB - This artical mainly introduces the choice of the fire-proof material, the causes of burning and the technical ensurement of fire prevention in the hyperbaric oxygen chambers. PMID- 12552739 TI - [Treatment of chronic prostatitis with computerized herbal drug penetrator]. AB - A computerized herbal drug penetrator for chronic prostatitis was designed and developed on the basis of a combination of the authors' clinical practice with modern Traditional Chinese Medicinal theories. The device has integrated many hi techs of current engineering science. It works by applying special herbal medicine, electro-magnetic effects, thermal moxibustion, perfusion and pulsated massage with water-capsule. The result of using this technique in the treatment of 100 cases has demonstrated its satisfying effectiveness. PMID- 12552740 TI - [Study on correlation between in vivo and in vitro tests of biomaterials- correlation between leukocyte chemotaxis test and muscular implant test in rabbits]. AB - The purpose of this study was to investigate the correlation between the results of in vivo and in vitro tests of biomaterials. With muscular implant test and leukocyte chemotaxis test, the levels of tissue toxicity of seven biomaterials were evaluated. The samples with high-level toxicity showed weak leukocyte chemotaxis, which would influence the anti-inflammatory reaction to foreign bodies. By the method of Spearman rank correlation, the coefficient was between 0.89286-0.98198 indicating that the correlation of the two tests is excellent. PMID- 12552741 TI - [Effects of chitin and chitosan on degradation of DL-polylactide in vitro]. AB - The authors assessed the variations of pH-values of the physiological saline used for in vitro degradation tests of DL-Polylactide (DL-PLA)/chitin and DL PLA/chitosan composites and DL-PLA. The weight-loss rations, the appearance and morphology of the tested specimens, and the molecular weights of DL-PLA in composites and pure DL-PLA were alse investigated. The results showed that both chitin and chitosan had significant inhibitory effects on the degradation of DL PLA, and the effect of chitosan was stronger than that of chitin. A discussion on the mechanism of such inhibition is presented. PMID- 12552742 TI - [Study of cytotoxicity of bioprosthetic heart valve material and its store solution]. AB - This study compared the cytotoxicities of bioprosthetic heart valve materials crosslinked by glutaraldehyde, stored in 4% formaldehyde or Hank's solution. Human embryonic pulmonary fibroblasts or L-929 cell culture in vitro were used. Cell proliferative inhibition index(CP II) was calculated for bioprosthetic heart valve materials using different store methods in different rinse periods(before, 10 days, 20 days, 30 days). The results demonstrate: (1) bioprosthetic heart valve materials stored in 4% formaldehyde or in Hank's solution both have significant cytotoxicity, and the longer the rinse time continues, the lower the cytotoxicity declines; (2) HEL cell is more sensitive than L-929 cell in detecting the cytotoxicity of toxic biomaterials but for weak toxic biomaterials the two cell lines are not significanth different; (3) formatldehyde solution increases the cytotoxicity of biomaterials stored in it, but the enhanced cytotoxicity can be easily relieved by rinse. The authors conclude that bioprosthetic heart valve materials have long term significant cytotoxicity and the biomaterial cytotoxicity test using human fibroblasts is more sensitive and precise than other tests. PMID- 12552743 TI - [Effects on the amount of total hemolytic complement levels(CH50) and immunoglobulin in serum induced by the implantation of biomaterials into rats]. AB - Three kinds of biomaterials, PES, YP and SR, were implanted into the subcutaneous tissue of rats and stayed there for 7, 14, 30, 60 and 90 days. Then using the immunity assaying system (Beckman, ICS) and the standard method of measuring the value of total hemolytic complement levels (CH50), we determined the values of immunoglobulin and CH50 in serum of rats. The results showed the amounts of IgG and IgA of rats with the implanted YP and SR materials were significantly higher than those of the control groups, and their value of CH50 was significantly lower than that of the control group. There was no significant difference between the PES group and the control group. PMID- 12552744 TI - [The properties of new affinity adsorbent for adsorbing the cycle immune complex in blood]. AB - This study was designed to select an affinity adsorbent from 5 kinds of adsorbents available. The adsorbent AA3 was found being specific to the adsorption of pathologic cycle immune complex (CIC). The physical properties such as specific surface area, proe volume and average pore diameter of adsorbents were determined. A discussion on the factors that affect the adsorption of CIC is also presented in this paper. PMID- 12552745 TI - [Study on heparin-like materials--sulfonization and blood compatibility of PVA, PET and PEU]. AB - In this paper, physical mechanical properties and changes of chemical structures of some heparin-like biomaterials were studied before and after sulfonization of PVA, PET and PEU. The results showed that these materials were easy to be sulfonized, but it was necessary to keep actual sulfonization degree for their mechanical properties. Platelets adhesion experiments demonstrated that the materials adsorbed very few and showed heparin-like anti-coagulant property after sulfonization. PMID- 12552746 TI - [Effects of fluid shear stress on neutrophil surface expression of adhesion molecules]. AB - Fluid shear is a physiological and functional environment for circulating neutrophils where various chemoattractants and cytokines have different effects on neutrophils, activating them or evoking inflammation. So far there have been abundant in vitro researches demonstrating that while neutrophils are activated, some adhesion molecules, which are expressed on the surface of neutrophils, are necessary to cell-to-cell adhesion, and the quantity or quality of these molecules will be changed. The above results were all obtained under static fluid which is quite different from physiological condition. Recently, we used Low Shear 30 to provide steady or sinusoidal-oscillated fluid shear stress on separated neutrophils with or without f-MLP/TNF stimulation. FITC-anti CD18 monoclonal antibody and PE-anti CD62L monoclonal antibody were added to label CD18 and CD62L on neutrophils suface respectively. Flow cytometry was used to quantify surface expression and positive cell percentage of these two adhesion molecules. The results demonstrat that fluid shear stress increases the expression of CD18 significantly and decreases the CD62L positive cell percentage, but it does not influence the CD62L expression on the CD62L positive cells. The effects of the different patterns of fluid shear stress are not much different, f-MLP or TNF can slightly decrease the expression of CD18 and make CD62L shedding respectively. When one of the two activators and the fluid shear stress act together, the expression of the two adherent molecules can be added. This indicates that the mechanical and biochemical stiumlative action can produce different effects through different receptors and different signal transduction pathways. So the mechanical environment should be considered in the research of activation and adhesion of neutrophils. PMID- 12552747 TI - [Changes of biomechanical properties in goats at different times after ovariectomy]. AB - This paper presents the changes of biomechanical properties in goats at different times after ovariectomy. 14 goats were randomly divided into three groups. These goats were ovariectomized and sacrificed at 180 days (n = 4), 360 days (n = 5) and 540 days (n = 5) after the operation. The right femur, tibia and metatarsus were taken out. The structure mechanical properties of femora were measured with the three-point bending method and those of metatarsus were measured with the compressing method. The material mechanical properties of flexure, compress and tensile strength in the compact bone specimens of femora and tibiae taken from broken ends were also measured. The results showed that the one-year group, compared with the half-year group, had all the bone geometric parameters decreased, but not statistically significant (P > 0.05); the elastic modulus of three-point bending of femora decreased by 23.27% and the others had no significant difference (P > 0.05); all the material parameters of flexurel, compress and tensile strength in compact bone specimens decreased, but only the ultimate strength of tibiae of flexure, compress and tensile strength decreased obviously (P < 0.05). The one and a half year group, compared with the one year group, had the bone density of femora and tibiae decreased, but not statistically significant (P > 0.05). All the bone geometric parameters of tibiae and the thickness of bone of femora increased (P < 0.05). The break load of three-point bending of femora increased (P < 0.05). Probably, it was related to the increase of the body weight in the one and a half year group. The elastic modulus and the ultimate strength decreased, but not statistically significant (P > 0.05). When compact bone specimens were used, the ultimate strength and elastic modulus of femora and tibiae in compress and the elastic module of tibiae in flexure decreased obviously (P < 0.05); and the others had no significant changes (P > 0.05). This study indicated that the degree of osteoporosis in ovariectomized goats increased at half a year, one year, and one and a half year postoperation, and hence suggested that the ovariectomized goat be a pretty useful big animal model of osteoporosis. PMID- 12552748 TI - [Effects of immobilization on the mechanical properties of rabbit patellar tendon]. AB - The aim of this study was to evaluate the effect of immobilization on the mechanical properties of rabbit patellar tendon(PT). IN this study, the rabbit knee was immobilized in extension for 6 weeks, then the mechanical properteis of PT were investigated. The results showed that the stress-strain relationship of PT could be expressed with exponential functions, and there were significant differeces between the normal group and the immobilized group in their theoretical curves of stress-strain relationship and two material constants of PT. Also, significant differences were found in tensile strength and tangent modulus between the two groups, but the maximum load of PT in the immobilized group did not decrease significantly. On the basis of the findings listed above, it is believed that PT would remodel and change its mechanical properties in response to the changes of mechanical stress exerted on it when the knee is immobilized. PMID- 12552749 TI - [A measurement and analysis on the dynamic changes of wall shear stress in altered arterial flow]. AB - This study was designed to estimate the dynamic changes of wall shear stress in reduced arterial flow and hence to get a better understanding of its effect on the endothelial adaptive remodeling. Left common carotid arteries of experimental rabbits were ligated distal to the origin of the thyroid artery to reduce flow by about 90% in the carotid upstream of this branch, and the vessels were examined respectively at different time intervals. The changes of mean shear stress showed three different stages. The shear stress decreased significantly during postoperative 1 hour to 7 days, increased markedly after 7 days and returned to control levels after 30 days. These changes were closely related to the decreased arterial diameters. The results indicate that the shear stress may paly an important role in the arterial remodeling after alterations in blood flow. PMID- 12552750 TI - [On the biomechanical reasons why children are liable to have fracture in supracondyle of humerus]. AB - With the viewpoint of biomechanics in theory, this paper analyses why children are liable to have fracture in supracondyle of humerus as a result of falling to the ground with the upper limbs propping up the body while adults usually have fracture in the middle or upper part of humerus under the same condition. The focus of discussion is the change of front-inclination in the lower end of humerus in the process of bone maturity from childhood to adulthood. PMID- 12552751 TI - [Effects of temperature on the vasoconstrictive force of norepinephrine in rat aorta]. AB - This study aimed at the influence of temperature on the vasoconstrictive effect of norepinephrine on rings of rat aorta and at the cause of the change observed. At the temperatures of 39.5 degrees C, 36.5 degrees C and 33.0 degrees C respectively, the vasoconstrictive force-time curve induced by norepinephrine and the equation of function were fitted. Statistical evaluation of the data was performed by F test. The difference was statistically significant (P < 0.01) among the three groups for relation constant X1 of function equation, but no significant difference (P > 0.05) was noted among the three groups for relation constant X2. Our experiments demonstrated that norepinephrine-induced vasoconstrictive force, in rat aorta, was obviously modulated by temperature. As the temperature of rat aorta was raised, the vasoconstrictive force and vasoconstrictive speed increased, and the effects of norepinephrine on vasculature were raised. The results suggest that the constant X1 for vasoconstrictive force is a factor sensitive to temperature. PMID- 12552752 TI - [Design of CXLY-II low frequency magnetic fields pulse generator]. AB - An experimental device, low frequency magnetic fields pulse generator, has been designed and accomplished in our department. It can provide low frequency magnetic fields pulse with the intensity from 150mT to 800mT and the frequency from 0 to 100Hz. This device is easy to operate and performs reliably. It can stably work for a long time and has been successful used in the experiment of magnetic biological effect. PMID- 12552753 TI - [Computer-assisted identification of fundus lesions and a study of its reliability]. AB - This study aimed at the use of computer for identifying fundus lesions quickly and reliably. We adopted the 'area filling' method to program in Visula C++, used the program to mesure the retina cotton-wool patches in diabetics (15 eyes), and then calculated the coefficient of variation to evaluate its reproducibility. The results showed that the coefficient of variation of the measurements ranged from 3.22% to 9.46%. In conclusion, this method is convient, quick and reliable. PMID- 12552754 TI - [Detection of P wave through wavelet transform and neural network]. AB - The ECG signal is decomposed throught wavelet transform, and then P of the ECG signal wave is detected by neural network. This method has been used as a supplement with satisfactory result. It is of importance to HRV analysis. PMID- 12552755 TI - [A new algorithm for direct volume rendering of medic image series]. AB - In this paper, a new algorithm based on Voxel Model is proposed. This algorithm combines the advantages of surface rendering and direct volume rendering. In the process of 3D reconstruction, OpenGL graphic standard and hardware accelerator can be used. By doing so, better reconstruction, result and fast speed can be achieved. Also, it is simple to turn this algorithm into programs. PMID- 12552756 TI - [Texture analysis of B-scan image using fractal dimension]. AB - Based on the texture analysis of B-Scan image fractal model, this paper presents the liver clinical B-Scan images and their fractal characteristics. It describes the semi-fractal characteristics of B-Scan images and calculates the fractal dimension(D) of the liver B-scan images of 10 normal persons, 3 patients with cancer of the liver, and 3 patients with cirrhosis of the liver. The results indicate that Fractal Dimension (D) is an important parameter in differentiating some diseases. On the basis of the influence of Fractal Dimension (D) and scale (epsilon), this paper suggests the texture classified parameter S. The experiment results indicate that the parameter S can be used to differentiate diseases efficiently. PMID- 12552757 TI - [Implementation of gateway between WWW and DICOM]. AB - This paper discusses the necessity of gateway between WWW and DICOM, shows the method of the implementation and the result. It brings forward a new way in which doctors interact with PACS and medical image device such as CT, MRI, ultrasonic, etc. It points out that the gateway between WWW and DICOM is a kind of transition between appropriative standard and general standard. This kind of transition will bring convenience to telemedicine information system and integrated medical information system. PMID- 12552758 TI - [Application of power band graph method to the modeling and simulation of cardiovascular system]. AB - This paper presents a computer simulation model of the cardiovascular circulation system, which describes the blood flow dynamic law in the cardiovascular system by the state equation. The model can be used in physiological study and computer aided medical education. In this paper, the Power Band Graph (PBG) modeling method is used to realize modeling of the human circulation system and conduct a simulation study on a simplified physiological system model. The results demonstrate that the PBG method, as an understandable and unity modeling method, is quite effective and practicable and can be used widely in the field of physiological system simulation. PMID- 12552759 TI - [Multiresolution analysis based denoising algorithm for dynamic esophageal pH monitoring signal]. AB - A denoising algorithm for dynamic esophageal pH monitoring signal is first introduced in this paper. The algorithm was developed on the basis of multiresolution analysis. The pH monitoring signal was investigated in experiments, then the algorithm was formulated by multiresolution analysis based on discrete dyadic wavelet transform combining with the results of the investigation mentioned above. Application of this algorithm to clinical data processing has proved its satisfactory denoising effect. PMID- 12552760 TI - [Effects of anticoagulants, temperature and sex on the spontaneous activation of neutrophils in vitro]. AB - From the view of cellular morphology and enzymic histochemistry, we used the pseudopod method and NBT test to measure spontaneous activation of neutrophils in vitro. The effects of anticoagulant, temperature, and sex on the percentage of spontaneously activated neutrophils were studied. The results suggest that heparin is a better anticoagulant than EDTA because EDTA because EDTA inhibits neutrophil activation; that a higher percentage of activated neutrophils can be obtained when incubated at 37 degrees C, compared with the percentage at 25 degrees C, and at 37 degrees C the highest percentage appears an hour earlier; and that females show a higher percentage of spontaneously activated neutrophils than males, and females' highest percentage appears an hour earlier than males'. These are of referential value for further studies. PMID- 12552761 TI - [A study on ovariectomized goat osteoblast culture in vitro]. AB - Osteoblasts were isolated from the frontal parietal bone of ovariectomized goat and were confirmed by testing for multiple biological features. The results showed that the goat osteoblasts cultured had the typical morphological feature of osteoblasts, the activity of alkaline phosphatase, and the ability of calcification in vitro. Also, the results proved that these goat osteoblasts in vitro had the same function and features as those in vivo. The experimental techniques in this study have provided a useful system for researches in bone metabolism. PMID- 12552762 TI - [The influence of pulsed magnetic fields on SOD's activity and MDA value in metabolism of mice]. AB - This experiment aimed to investigate the influence of pulsed extremely-low frequency-magnetic fields on free radical metabolism of mice. Thirty-two mice were randomly divided into four groups and were exposed to 0 T, 0.25 T, 0.34 T and 0.64 T intensity pulsed magnetic fields of 20 Hz for 40 min. The 0 T exposed group was the control group. The free radical metabolism, SOD's activity and MDA, of mice were measured respectively. The result showed the SOD's activity and MDA of the 0.34 T exposed group were both significantly lower (P < 0.01, P < 0.05) than that of the control group, while the 0.25 T and 0.64 T exposed groups were not significantly different from the control group. This demonstrates that the biological effect of pulsed extremely-low-frequency magnetic fields on free radical metabolism of mice varies and depends on the intensity of the fields. The "window" effect may exist. PMID- 12552763 TI - [Expression and significance of neuropeptide and neurotensin in the extract of platelet of chronic renal failure patients during hemodialysis]. AB - The aim of this study was to investigate the change of neuropeptide Y(NPY) content and neurotensin (NT) content of the platelet from chronic renal failure (CRF) patients during hemodialysis (HD) and explore its mechanism of action. Platelet was separated from the patients' plasma and the NPY and and NT contents of the platelet and plasma were dynamically observed pre-HD and post-HD. 30 healthy people were chosen as the controls. The results showed the NPY and NT contents of the platelet extract were (58.18 +/- 21.29) ng/10(9) and (25.38 +/- 13.43) ng/10(9) respectively. The NPY content of the platelet extract of the CRF patients was obviously decreased and the NT content of the plasma was obviously increased however, the NT contents of the platelet extract and the plasma were both higher than those in the control group. The NPY content of the platelet extract and NT content of the plasma at the post-HD obviously increased as compared with pre-HD, but the NT content of the platelet extract and the NPY content of the plasma obviously decreased. There was an obvious correlation between NPY and NT. The authors conclude that during the period of CRF, the immunoreactions which mainly involve bio-active substances such as NPY, NT and 5 hydroxytryptamine released by the platelet to interfer the vasoconstrictive effect are the main pathologic factors to induce renal hypertensin and renal vasospasm. PMID- 12552764 TI - [Advances of research on speech signal processing strategy in cochlear implant]. AB - Cochlear implant is a new technology to restore hearing ability to totally deaf people. First of all, this paper presents its principle and system design. Then, the advances of research on its speech signal processing strategy is reviewed in detail. Finally, on the basis of preliminary results, further promising development in cochlear implant is proposed. PMID- 12552765 TI - [Vroman effect of plasma protein adsorption to biomaterials surfaces]. AB - Fibrinogen adsorption from plasma to biomaterials surfaces passes through a maximum when studied as a function of adsorption time, plasma concentration, or column height in narrow spaces and these are called Vroman effect. Studies have demonstrated that Vroman effect is a general phenomenon of plasma proteins and reflects the competitive adsorption of plasma proteins for a limited number of surface sites. In this paper, the factors affecting Vroman effect, the relationship between contact activation and fibrinogen displacement, and the significance of the Vroman effect with respect to blood-material interactions are reviewed. PMID- 12552766 TI - [Progress in technique of computer for three dimensional reconstruction from biological tissue series slices images]. AB - The development of the technique of computer processing images and biotechnology from late 80's to early 90's has rovided the possibility of displaying three dimensional images of biological tissues truly. Now the technique of computer for three dimensional reconstruction is more and more widely used in the biological territory. This paper introduces the progress in the technique of computer for three dimensional reconstruction from biological tissue series slices images. Emphasis is laid on registration, reconstruction & display, image data compressing and image automatic segmentation. PMID- 12552767 TI - [Recognition and classification of heart sounds]. AB - This paper reviews the methods and research advances of recognition and classification of heart sounds. The methods include the statistical techniques and artificial neural network. It also discusses the main problems in the methods. PMID- 12552768 TI - [Computer-aided teaching system of heart sound]. AB - This paper details a full set of plan of the computer-aided teaching of heart sound. By means of digital sound technology one is capable of playing realistically heart sound of various kinds based on database. Each heart sound is demonstrated by a proper explanation of its characteristics and a heart sound graph. This method gets over the shortages of traditional teaching methods. Besides, this system can be used to test users' level of auscultation. PMID- 12552769 TI - [Development of continuous rat infusion device]. AB - In order to perform better scientific researches on fluid resuscitation, total parenteral nutrition and continuous drug administration in rats, we developed a set of rat infusion device. The device, the characteristics of reasonable design, simple operation, low cost, good practicability, and making it by oneself or buying a whole set, can not only assure rats to run around, drink water, eat food freely, but also insure continuous infusion and drug administration during the experimental study of rats. PMID- 12552770 TI - [Computer-assisted measurement of mean transit time and partial mean transit time]. AB - A pulmonary function measurement system was developed. This system was made up by Fleisch flow transducer, the pressure transducer and 486 computer. It can calculate and print out the mean transmitted time (MTT) the partial mean transmitted time (MTTp) and the routine pulmonary function automatically. PMID- 12552771 TI - [Preparation of chitosan-gelatin mixed film and study on it's biodegradability]. AB - A chitosan-gelatin film is prepared and the biodegradability and biocompatibility of the film are investigated by animal experiments. Results suggest that chitosan gelatin film may be biodegraded faster and it has good biocompatibility. Lysozyme may promote biodegradation of the mixed film. PMID- 12552772 TI - [Tetracycline tracing of artificial bone composite with high biological induction in repairing bone defect]. AB - The tetracycline tracing method was adopted to study the new bone formation at the early stage of two different composite artificial bones (HA-bBMP-Co and HA Co)in repairing segmental bone defect in 8 rabbits. The results indicated that from the early stage even just one week after implantation, HA-bBMP-Co acted much faster than HA-Co did in inducing new bone formation, suggesting that HA-bBMP-Co be of good quality in ferms of histo-compatibility and osteoinduction. PMID- 12552773 TI - [Design and study of carbon fiber tracheal prosthesis]. AB - 32 healty adult dogs were selected for this experiment. 10 of them were subjected to the tracheal biomechanics test using indices including the relation between stretcher ratio (lambda) and stress (T), the squeeze stress (delta jy) of medical silk thread on trachea, the side stress (Ts) inducing the tracheal collapse, the functional maximum angle (psi max) of tracheal, and the sever area torsion angle (theta max) of tracheal functional maximum curved. According to the indices measured, two types of tracheal prosthesis were designed, and were made of carbon fiber and silicon. They were the straight tube type tracheal prosthesis and the bifurcate type tracheal prosthesis. The straight tube type tracheal prosthesis was studied with a design of two groups comprising a total of 11 dogs. In the experiment group (n = 6), the outer surface of the tube was not coated with silicon, the average survival period was 379.8 days. In the control group (n = 5), the outer surface of the tube was coated with silicon, the average survival period was 90.4 days. The bifurcate type tracheal prosthesis was studied in 11 dogs, the average survival period was 4.32 days. The main causes of death in the experiment were infection and anastomotic dehiscent. PMID- 12552774 TI - [The curvature design on artificial femoral stem in Chinese population]. AB - Based on A-P view geometory and lateral view geometory X-ray Photograph, the upper geometric curves of intermedulla were measured from each bone in 84 femur specimens in Chinese population. All curves were parameterized by lest mean square method. A statistical analysis on parameterized data was made for finding out the growth law of femur in Chinese population. The shape of the femur is classified into 7 types. This results of the research is for the the design of Chinese-Type artificial femur. PMID- 12552775 TI - [Design and experimental study of an opening type implantable artificial pancreas equipped with a pulse pump and a micropore slow delivery pump]. AB - According to physiologic character of insulin secretion of normal body, one new design has been advanced. It can provide insulin with both pulsed and slow released ways, and imitate separately two completely different forms of insulin secretion. Through extracorpored experiment, one parallel model has been set up. One function is similar to post-cibum largely and quickly pulsed insulin secretion, other one is continuous and stable micro-secretion in 24 hours. The theoretical base for researching furtherly the open implanted artificial pancreas (pump) depends on the result of this article. PMID- 12552776 TI - [A study of affinity adsorbent AA-3 in cleaning the cycle immune complex of rabbits]. AB - Conducted a hemoperfusion (HP) experiment on AA-3 for the treatment of type III allergy in rabbits and observed the change of Cycle Immune Complex (CLC) and the blood mutual adaptation (BMA). The results showed that the CIC level was 195.52 +/- 12.79 ug/ml before adsorption and 119.01 +/- 10.4 micrograms/ml after ansorption, the adsorption rate being (39.13 +/- 2.19)%. The experiment also demonstrated satisfactory BMA during AA-3 HP. PMID- 12552777 TI - [The effect of pressure on the morphology of superior mesenteric vein in rabbits]. AB - The effect of pressure on the morphology of the superior mesenteric vein of rabbits (SMV) was researched with the method of constant pressure fixation. The results indicate the morphology of SMV is similar to human umbilical cord vein. It looks like the structure of femoral artery of vein in many ways. Under constant pressure fixation (4 kPa), the internal elastic membrane of SMV is straightened, and the wall of SMV becomes thin, with its media thinning from 51.680 microns to 32.3260 microns. The long diameter of nucleus of the smooth muscles decreases from 1.9146 microns to 3.4980 microns, the short diameter of nucleus of the smooth muscles becomes from 0.7884 micron to 0.5228 micron. The direction of the tension on the smooth muscles is mainly longitudinal. PMID- 12552778 TI - [Physical simulation of radial artery displacement]. AB - A previous developed physical cardiovascular simulation is used to investigate the displacement of radial artery during a cardiac cycle. Simulation result shows that the displacement is much greater than the change in the radius of the radial artery. The result is similar to the clinical observation reported before. Based on the simulation result, it is suggested that the displacement of radial artery plays an important part in the Maixiang (pulse condition), which is one of the most important contents of diagnostics in Traditional Chinese Medicine. PMID- 12552779 TI - [Changes in blood constitutive equation parameters during shock in rabbits]. AB - Based on the method which can more precisely obtain the constitutive parameters, we studied the changes of hemorphelogical characteristics of rabbits during shock and the effect of MN9200, a king of calcium channel anatogist injected into the veins of the rabbits. The results showed that shock induced a significant change in hemorheological index and NM9202 improved the condition. PMID- 12552780 TI - [A human knee articulate mathematical model on femur-tibia-patella 3-segmetents]. AB - In this paper, based on the previous work of S. Turgut Tumer et al, a human knee articulate mathematical model on femur-tibia-patella 3-segments is established by introducing patella-femur joint. This model includes both rolling and slipping motions of femur-tibia joint and femur-patella joint. It can be employed in simulating the biodynamic response of human lower extremity. PMID- 12552781 TI - [A physical model for cuff blood pressure measuring technique and its use in the systolic blood pressure measurement]. AB - The traditional blood pressure measurement by oscillometric method is based on statistics theory, so individual error always takes place in measurements. In this paper, we devise not only a technique that can acquire cuff pulse waves undistorted, but also a physical model of cuff measuring process. Using undistorted data and based on this model, we present a new technique for systolic blood pressure measurement. It has been proved to be more accurate than the traditional method. PMID- 12552782 TI - [Determination algorithms in oscillometric blood pressure measurement]. AB - This paper discusses the method of oscillometric blood pressure determination. A new criterion for systolic pressure and diastolic pressure determination has been proposed, which is based on ratio changes of the oscillation amplitude under a wide normalized range. Primary experiment results show the new criterion is effective. PMID- 12552783 TI - [Function of modified YSI membrane in glucose sensor]. AB - This study was designed to improve the stability, accuracy and interference resistance of the glucose sensor system and to fomodify the correlation between electrode current and glucose concentration. Catalase was immobilized in YSI membrane. Experiments in vitro were conducted in different concentrations of glucose or ascorbic acid or paracetomol. The results showed the background current, 0.37 +/- 0.06 nA; the drift values in 0 mM G.S. and 10 mM G.S. were (0.20 +/- 0.14)%/30 hr and (3.22 +/- 0.40)%/30 hr respectively. The mean value of drift in 10 mM G.S. was 0.103% mM-1hr-1. The correlation between electrode current and glucose concentration gave satisfaction (r = 0.986); it ranged from 0 mM G.S. to 20 mM G.S. Lower concentration of ascorbic cid could not interfere n the glucose sensor system; higher concentration of ascorbic acid and paracetomol only produced very low interference current. These suggest that the modified YSI membrane contributes greatly to a stable, accurate and reliable glucose sensor system. PMID- 12552784 TI - [A methodological study on the detection of myocardial ultrasonic integrated backscatter]. AB - The cyclic fluctuation of ultrasonic integrated backscatter(IB) of myocardium has been found to be a potential indicator of clinical significance in the diagnosis of myocardial ischemia. To investigate the feasibility of such a technique, a clinical IB testing system is established in our laboratory. Six male subjects with no history of heart diseases were tested by using this system. Based on the clinical data obtained during the tests, the effects of echo position, bandwidth, and different calculation algorithms on IB value were analyzed and compared. The results are available to provide a guideline for the design of instrumentation for IB detection. PMID- 12552785 TI - [An optical model of axial reflection and its clinical application]. AB - A developed optical model of axial reflection on retinal blood vessels has been presented in this paper. The loss of the transparency of retinal arterial vessel wall is a frequent expression of retinal vascular pathology. With the change of parameters, the model can simulate the progress of the axial reflection exactly during the onset and progression of hypertension. This non-invasive and quantitative method is useful for study in the relationship between hypertension and retinal vessel axial reflection. It can help doctors approach a diagnosis of hypertension. PMID- 12552786 TI - [Establishment and discretization of human torso computer images for ECG simulation]. AB - The establishment and discretization of human torso computer image is an important aspect in studying of ECG simulation. A rapid and effective method is presented in this paper, which establishes and discretes image of human torso exactly by curve fitting and by introducing concept of "nihility point". It also has some value for solving problem of discreting field region in numerical calculation of electromagnetic field. PMID- 12552787 TI - [An automatic method segmenting lung regions from digital chest radiograph]. AB - It is difficult to process digital chest image with ordinary method because of the wide dynamic range of chest radiograph. Anatomically adaptive processing is a better method for chest image, but it needs automatic segmentation of the image. In this paper, an effective method for segmenting lung regions from digital chest radiograph is presented. This method automatically calculates the lung/mediastinum-subdiaphragmatic threshold according to the histogram of chest image processed. PMID- 12552788 TI - [Self-organization neural network based ultrasonic heart image segmentation]. AB - Segmentation is one of the most difficult problems in multidimensional reconstruction of ultrasonic heart image. In the present paper, a segmentation method of ultrasonic heart image using the self-organization neural network has been studied. This is an unsupervised segmentation method, which can segment images through clustering automatically. The results show that the present method has significant benefits over the traditional K-means algorithm. PMID- 12552789 TI - [Crackle detection and classification based on matched wavelet transform]. AB - In this paper, we present a method for crackle detection which is based on 'matched' wavelet transform. We first modeled crackles as a mathematical function. Then we designed a matched mother wavelet based on this model. Applying a soft-threshold to the results of the continuous wavelet transform to suppress noise further, we obtained the optimal scale Crackles were detected based on the envelope of the signal at optimal scale, and could be classified based on energy distribution with scale. The theory, methods and experimental results are given in detail. PMID- 12552790 TI - [Development of a long-term dynamic ECG monitoring system (Holter system)]. AB - The development of a Holter system is introduced here with the emphasis on some key problems such as low power consumption hardware design, high efficient real time algorithm, high speed reliable playback interface and full function interactive software. PMID- 12552791 TI - [Surface phenotype on dendritic cells and their immunological significance]. AB - Dendritic cells (DC) are very important antigen presenting cells(APC) in immune response. DC gradually gets mature while it migrates in vivo. Compared with mature DC, immature DC plays a different role in immune response. In this review, the recent advances in researches on specific cell surface marker and costimulatory molecules on DC as well as their immunological significance and methodology of propagation in vitro are discussed. It may be beneficial for further study of DC. PMID- 12552792 TI - [Surface treatments for dental titanium castings]. AB - Because pure titanium is highly reactive at high temperature, its casting surface is easily coated with a thick contaminated layer that affects its physical and chemical properties, and consequently a stable superficial oxide membrane develops in the presence of air or water even at room temperature, which makes the surface discolored. Therefore, surface treatments for pure titanium castings are more important than those for other dental alloy castings. This article reviews the methods of surface treatments for dental titanium castings in four aspects: physical finishing, chemical finishing, electro-chemical finishing and surface modification technology. PMID- 12552793 TI - [Progress in the controlled release systems of hirudin]. AB - Hirudin is very effective in treating and preventing cardiovascular disorders. In this review several polymeric controlled release systems of hirudin are introduced. The advantages of the local delivery for restenosis and the feasibility of endovascular drug-containing pharmaceutic stents for local therapy of cardiovascular thrombosis are discussed. PMID- 12552794 TI - [Measurement of forces between biomolecules by atomic force microscopy]. AB - The intermolecular forces of the biological samples may be measured by atomic force microscopy, which has provided a powerful means for us to understand the principles of living phenomena. Some systems in which the interaction forces between biomolecules have been measured hitherto include: 1. ligand-receptor; 2. actin-myosin; 3. proteoglycans; 4. DNA complementary strands and nucleotides; 5. special organic chemical groups. A literature review on these objects is presented in this article. PMID- 12552795 TI - [The development and application of telemedicine]. AB - Telemedicine, which began in the late 1950s, is the combination of modern technologies such as medicine, digital communications, computer and multimedia. The study of telemedicine in some developed countries has made great progress in 1990s. However it began somewhat late in our country. This article reviews the development of telemedicine and its application. The authors also point out some problems in telemedicine that need to be solved. PMID- 12552796 TI - [The research fields and advances of biomedical engineering]. AB - The research fields and advances of biomedical engineering have been reviewed in four aspects, biomedical material, biomedical engineering appliance, tele diagnostic system and biomedical recovery engineering with 14 references. PMID- 12552797 TI - [The different functions of glnB and glnZ from Azospirillum brasilense YU62 in the control of nitrogen fixation]. AB - The glnB and glnZ genes of A. brasilense have 70% homology at nucleotide sequence. glnB is located in a 3.7 kb Eco RI+ PstI fragment and glnZ is located in a 3.7 kb SalI fragment. Both glnB and glnZ genes were mutagenized by Kmr cassette insertions and glnB- and glnZ- mutants were obtained. glnB- mutant did not have any nitrogenase activity, while glnZ- mutant still has nitrogenase activity. The coding regions of glnB and glnZ were cloned into pVK100 vectors and recombinant plasmids pVK-II and pVK-Z were obtained, respectively. The recombinant plasmids pVK-II and pVK-Z were introduced into glnB- and glnZ- to produce C-glnB and C-glnZ, respectively. C-glnB can restore nitrogenase activity and C-glnZ does not have effect on nitrogenase activity. When pVK-II and pVK-Z were introduced into A. brasilense Yu62 and draT-, respectively, the Yu62-II (containing pVK-II) and draT-II (containing pVK-II) have higher nitrogenase activity than that of wild type Yu62. In contrast, Yu62-Z (containing pVK-Z) and draT-Z (containing pVK-Z) has no effect on nitrogenase activity. The nifA(-)-II (containg pVK-II) and nifA(-)-Z (containing pVK-Z) still have no nitrogenase activity. PMID- 12552798 TI - [Studies on the genome size and structure of Gluconobacter oxydans SCB329]. AB - In the "Two-step fermentation" of Vitamin C synthesis, Gluconobacter oxydans SCB329 is responsible for the production of 2-keto-L-gulonic acid (2-KLG), which is an important precuror of vitamin C synthesis. The intact chromosome was prepared from logarithmic phase cells by agaraseembedded method and was analysized by restriction endonucleases and contour-clamped homogeneous electric field pulsed-field gel electrophoresis (PFGE). Spe I (5-ACTAGT) produced 24 fragments, ranging in size from 10 to 320 kilobases (kb). Xba I (5-TCTAGA) yielded 40 fragments (4 to 200 kb). A total genome size of approximately 2,700 kb was determined by summing the fragment length. Analysis of the entire genome of SCB329 by PFGE revealed that the genome of SCB329 consist of a chromosome which is 2,500 Kb in length and a large plasmid which is 245 kb. After linearization of the DNA by DNase I and S1 nuclease, in contrast with the band which can not be viewed, the band of chromosome and plasmid were appeared, this suggest that structure of the chromosome and the plasmid were circular. PMID- 12552799 TI - [Isolation and characterization of a plasmid pTS50, which encodes nisin resistance determinant in Lactococcus lactis TS1640]. AB - Three nisin-resistant Lactococcus lactis strains were screened from 197 samples of fresh milk on a selective medium (M17) supplemented with nisin, lactose and bromocresol purple, and were confirmed to have the nisin resistance determinant (nsr) by PCR amplification. Physiological and biochemical tests as well as Lactococcus lactis specific 16S rDNA sequence analysis revealed that the three strains all belong to Lactococcus lactis subsp. lactis. A large plasmid, pTS50, was identified in L. lactis subsp. lactis TS1640 which encodes resistance to nisin following electroporation of the total plasmids DNA into L. lactis MG1363. The molecular weight of plasmid pTS50 was estimated to be 47 kb by restriction analysis of BamHI, EcoRI, HindIII, NcoI, PstI and nsr was localized on a 1.9 kb EcoRI fragment by Southern hybridization. PMID- 12552800 TI - [The characterization of pJW566 from L. Lactis subsp. cremoris W56]. AB - The plasmid pJW566 was isolated from L. lactis subsp. cremoris W56, one strain for Danish chadder mixed starter cultures. The strain containing plasmid pJW566 showed resistance against three common phages species 936, c2 and P335 worldwide. It was found that pJW566 encoded for an restriction and modification system, and showed strong resistance to phage CHCP412 when it was introduced into the industrial strain L. lactis CHCC2281 in milk medium. The endonuclease activity analysis indicated that the endonuclease required Mg2+, ATP, and was stimulated by AdoMet. PMID- 12552801 TI - [Introduction of Vitreoscilla hemoglobin gene in a recombinant E. coli for PHB production]. AB - Vitreoscilla hemoglobin gene (vgb) was introduced into the recombinant Escherichia coli VG1(pTU14) for production of poly-beta-hydroxybutyrate (PHB), in order to enhance the oxygen uptake of strain from molecular level and to solve the problem of oxygen limitation in process of fermentation. The carbon monoxide difference spectra analysis of Vitreoscilla hemoglobin (VHb) showed that vgb could be successfully expressed in VG1 (pTU14), and this expression was under the regulation of dissolved oxygen (DO). The higher amounts of VHb could be got correspond to the lower level of DO. Because of the introduction of vgb, cell growth and PHB accumulation of VG1 (pTU14) were all strongly promoted during the process of batch culture. PMID- 12552802 TI - [Transfer and expression of tyrosinase gene of Pseudomonas maltophilia in Pseudomonas pseudoalcaligenes]. AB - Pseudomonas pseudoalcaligenes is a notably killing maggots bacterium, which was isolated from natural dead maggots in the manure pits in the countryside of Yancheng. It easily die from effect of ultra-violet ray when it is exposed in the sun. Antiradiation effect of melanin is quite strong. Mel gene of P. maltophilia AT18 has been introduced into P. pseudoalcaligenes, and enabled it the ability of producing melanin steadily. Southern hybridization studies confirmed that the small fragment cloned in the P. pseudoalcaligenes comes from P. maltophilia DNA. SDS-PAGE analysis also revealed that an additional protein of 18 kD, which was equal to the size of the putative tyrosinase according to mel fragment, was exressed in the P. pseudoalcaligenes recombinant carrying the mel gene, The results of assay show that antiradiation effect of recombinant is quites strong, the effect time of killing maggats is longer than the recipient, the recombinant can't infect animals and fowls. PMID- 12552803 TI - [Homologous recombination in Streptomyces lincolnensis B48]. AB - To study frequency and mechanism of homologous recombination in Streptomyces, an E. coli plasmid which cannot replicate in Streptomyces was transformed into Streptomyces lincolnensis B48. After homologous recombination between delta lincomycin biosynthetic genes inactivated by thiostrepton resistant gene (tsr) carried on pYYE04al and homologous sequences on the chromosome, S. lincolnensis YY1 and S. lincolnensis, YY2 were obtained on SMA with low thiostrepton concentration. Hybridization of chromosomal DNA samples of S. lincolnensis YY1, S. lincolnensis YY2, standard S. lincolnensis and S. lincolnensis YYc digested with SmaI with the probe of tsr gene gave signal corresponding to a fragment of 1.5 kb in the former two; Nevertheless, hybridization of chromosomal DNA digested with Hind III and Sma I using the probe of delta lacZ' gene resulted in positive fragment of 4.4 kb only in S. lincolnensis YY2. Southern hybridizations indicate that S. lincolnensis YY1 is the result of homologous exchange while S. lincolnensis YY2 comes from bomologous recombination. To prove the existence of E. coli replicon and ampicillin resistant gene on the chromosome of S. lincolnensis YY2, its DNA digested with SphI was ligated and then transformed into E. coli JM83 competent cell. Two transformants named pSLE1 grew on the plate containing ampicillin. It's confirmed that pSLE1 is a part of pYYE04a1 from its digestion with different enzymes. PMID- 12552804 TI - [Studies on in vitro expression for gI gene of Marek's disease in E. coli by pGEX vector]. AB - Glycoprotein I Gene(gI) was amplified from genomic DNA of Marek's disease virus (MDV) 648 strain by polymerase chain reaction(PCR). PCR product was cloned into pGEX-6p-1 according to the right open reading frame(ORF). The expression of GST gI fusion protein was studied in detail on many factors including temperature, timing and IPTG. The curve for OD600 and the growing time of the recombinant bacteria is also established., which is helpful to find the optimal inducing time. GST-gI fusion protein was identified by SDS-PAGE and Western-blotting., and the result shows that the best concentration of IPTG is 0.2-0.5 mmol/L and inducing time have great effects on expression while temperature has little. The fusion protein was injected into mouse five times to identify its antigenicity and the result is positive in indirect fluorescent assay IFA. PMID- 12552805 TI - [Genomic structure and proteins sequence analysis of full-length of segment A of three infections bursal disease viruses]. AB - The full-length of segment A of three infectious bursal disease viruses (IBDV), including an attenuated strain HZ2, an attenuated vaccine strain JD1 and a virulent field isolate ZJ2000, were cloned by long RT-PCR and sequenced respectively. All the three results revealed the identity of IB-DV with two overlapping open reading frames (ORF) flanked by 5'- and 3'-noncoding regions(NCR) in 3259 bp long. The strains shared high identity with each other at nucleotide or deduced amino acid level, and also had four unique sites H253, N279, T284, R330 which are common in other attenuated and some classic or highly virulent strains. The virulent strain ZJ2000 had several key amino acid mutations located in hypervariant region of VP2 and near the VP2-VP4 cleavage site of polypeptide, which is probably related to the virulence. Sequence comparison supported that VP2 is not the sole determinant of the virulence. The highly conservation in 5'- and 3'-NCR of different strains indicated the NCR may be not responsible for the virulence. But the same conservation appeared in VP5 revealed another complex relationship between VP5 and the virulence. PMID- 12552806 TI - [Expression of human leukemia inhibitory factor gene in Pichia pastoris]. AB - The 575 bp artificial synthesized human leukemia inhibitory factor (hLIF) gene was cloned into vector pPICZ alpha A and the constructed expression vector pPICZ alpha A-hLIF was linearized by SacI and transformed into Pichia pastoris X-33. After the screening for Mut phenotype and the PCR analysis of the transformants, hLIF gene expressed in Pichia pastoris. SDS-PAGE and Western blot analysis of the culture supernatants showed that hLIF gene could express as 58.5 kD protein with the desired immunogenicity. Density scanning of the SDS-PAGE gel revealed that the targeted protein accounted for 32.8% of the total protein in the supernatants. The expressed products can inhibit the clone formation of murine's teratoma cells. PMID- 12552807 TI - [Effect of 3'-UTR of EG I from Trichoderma reesei on its gene expression in Saccharomyces cerevisiae]. AB - Several industrial yeast are developed as ideal expression hosts for the production of the commercially useful proteins. The expression levels in yeast cells of the heterologous proteins are affected by the regulation factors of the genes themselves. The full-length cDNA coding for EG I from Trichoderma reesei, the cellulose-degrading filamentous fungus, was expressed in Saccharomyces cerevisiae H158. EG I produced by the recombinant S. cerevisiae exhibits maximal activity at 50 degrees C-60 degrees C, pH 5.0. It was observed that removal of the 3'-untranslated region (3'-UTR) from EG I cDNA resulted in no active EG I produced by recombinant yeast. RT-PCR analysis indicated that unlike the yeast cells harboring full-length EG I cDNA, there was no detectable EG I mRNA in the yeast cells harboring EG I cDNA without 3'-UTR. The data suggested that 3'-UTR is important for the expression of EG I in Saccharomyces cerevisiae. PMID- 12552808 TI - [Mapping the interaction site of Rpb2 and Rpb3 subunit of fission yeast RNA polymerase II]. AB - To map the interacting site of subunit Rpb2 to subunit Rpb3 of RNA polymerase II in fission yeast Schizosaccharomyces pombe, the yeast two-hybrid system was employed in this paper to screen the interacting clones between Rpb2 and Rpb3.4 fragments of Rpb2 cDNA were cloned into the Ga14 BD vector pAS2. The 4 clones were named as pAS2 Rpb2-1, 2-2, 2-3 and 2-4, respectively. The complete cDNA of Rpb3 was cloned into the Gal 4 AD vector pGADGH. The clone was named as pGADGH Rpb3. The two-hybrid plasmids pGADGH Rpb3 and pAS2Rpb2-1, 2-2, 2-3 or 2-4 respectively were cotransformed into host cell yeast Y190. The interaction positive cotransformants were identified by beta-gal activity assay. The beta-gal positive cotransformants were selected from pGADGH Rpb3 and pAS2Rpb2-4 two-hybrid system. DNA sequencing and alignment results showed that the interacting site of Rpb2 to Rpb3 located within the fragment from base 2701 to 2966 of Rpb2 cDNA, or within the C-termini polypeptide from amino acid 902 to 989 of Rpb2 protein. PMID- 12552809 TI - [Studies on the ribosomal RNA gene(rDNA) of a microsporidium isolated from Pieris rapae L]. AB - Nuclotide sequence (1205 bp) of small subunit ribosomal RNA (SSUrDNA) of a microsporidium isolated from Pieris rapae L. (abbr:MPr) was specifically amplified by polymerase chain reaction (PCR). Another fragment of 657 bp downstream of MPr SSUrDNA 3'end was amplified with two other primers. Within this 657 bp fragment, the putative 3'terminus of MPr SSUrDNA and the extreme 5' of large subunit ribosomal RNA gene. (LSUrDNA) were identified, which situated at base 145, 146-186 and 187, respectively. Then the full sequence of MPr ssurDNA is 1245 bp. Its GC content was also nearly 34%. The ITS region (internal transcribed spacer), positioned between the ssu and LSUrRNA genes, was found to be 41 bp in length. The LSUrDNA 580r region of MPr is 470 bp, longer than 437 bp of Nosema apis, 447 bp of Nosema algerae. The secondary structure of MPr SSUrRNA was constructed. These analyses of MPr rRNA gene contributed to the somewhat limited microsporidian taxonomic classification based on morphology. PMID- 12552810 TI - [Purification and some properties of D-hydantoinase produced by Pseudomonas 2262]. AB - A D-hydantoinase produced by Pseudomonas 2262 was purified to electrophoretic homogeneity by the steps of thermal treatment, (NH4)2SO4 fractionation and column chromatography with Q-Sepharose fast flow, phenyl-Sepharose fast flow and Superose 12. Purification of about 60 fold was achieved with an overall yield of 16%. The relative molecular mass of the native enzyme is 109 kD and that of subunit is 53.7 kD by the analysis of Native and SDS-PAGE as well as gel filtration respectively. Some properties of the enzyme such as the sensitivity to thiol reagent and the effects of metal ions, for instance inhibited by Zn2+ and activited by Mn2+, Mg2+ are identical to dihydropyrimidinase. The optimum temperature and pH for enzymatic catalysis are 70 degrees C and 8.0 respectively. The enzyme activity is stable under 60 degrees C and in the pH range of 6-10. The N-terminal sequence for 10 amino acid residues is MDKLIKNGTI. PMID- 12552811 TI - [Fermentation conditions for production of alkaline elastase by alkaliphilic Bacillus XE22-4-1]. AB - A extracellular alkaline elastase producing bacterial strain Bacillus XE22-4-1, with the optimum pH of 10.0 for its growth, was isolated and screened from alkali lake of Tibet. 2% glucose and 0.25% yeast extract are respectively the appropriate carbon and nitrogen source on elastase production. Soymeal can promote elastase production. The fermentation conditions in a 2 L stirred tank were investigated. The results have revealed that the dissolved oxygen is the most effective factor on elastase production. The maximum elastolytic activity reached 266 u/mL among 48 hours of fermentation by combination of enhancing aeration and changing the mixing speed. PMID- 12552812 TI - [Studies on characteristics of kinetics and metabolic shift of genetically engineered yeast Pichia pastoris in high-density chemostat cultivation]. AB - Kinetics and metabolic shift of DNA recombinant yeast Pichia pastoris was studied in high-density chemostat cultivation in which the glycerol was used as a limited substrate. The experimental results showed that (1) cell optical density (OD600) of Pichia pastoris was linear with its dry cell weight (DCW) and wet cell weight (WCW), and reached 100 OD600 = 128.3 g WCW/L or 100 OD600 = 22.9 g DCW/L; (2) The relationship of specific growth rate (mu) and residual concentration of glycerol is accorded with Monod Equation, to obtain mu = mumax S/(Ks + S), where mumax = 0.366 h-1, Ks = 0.1823 g/L. The maximum cell yield of glycerol substrate YG = 0.54 g/g, growth maintaining coefficient m = 0.0069 g/(g.h), The cell yield of oxygen YX/O2 = 30.96 g/moL, and the optimum theoretical dilution rate Dm = 0.341 h-1 by parameter inference; (3) With specific growth rate (mu) increasing, glycerol metablic flux shifts linearly from glycogenesis and hexosephosphate pathways to glycolysis and tricarboxylic acid cycle pathways on basis of variations of consumption rate of NH3.H2O and respiratory quotient value. PMID- 12552813 TI - [Construction of mucosal vaccine derived from HBV surface antigen epitope A and the assay of its immunogenicity]. AB - The fusion gene of HBV(adr) surface antigen epitope and B subunit of Cholera toxin was constructed and expressed successfully in E. coli at high yield. After denaturation-renaturation process, SDS-PAGE analysis showed that most of the renatured product reassociated in a pentameric form which was the same as natural CTB. Western blot analysis indicated that the immunogenicity of HBsAg antigen epitope was conserved. Moreover, ELISA analysis of the sera of orally, intranasally and intraperitoneally immunized mice showed that the circulating IgG antibodies to HBsAg were developed. The results may be helpful for constructing the novel mucosal vaccine with high efficacy. PMID- 12552814 TI - [Analys of outer membrane protein and drug-resistant of Edwardsiella tarda]. AB - The SDS-PAGE profiles of out membrane protein(OMP) of 27 Et strains showed diverse patterns and they could be divided into six types. Between 21 pathogenic strains and 6 non-pathogenic strains there were obviously difference. The pathogenic strains isolated from China belong to E type and were simary to ATCC 15947. The OMP bands of pathogenic strains were more and denser than those of the non-pathogenic strains, and the strains from same souce had the identical patters. Interestingly, the non-pathogenic strains from different sources also had the almost same OMP profiles. What's more, they were also high resistant to some antibiotics such as Smx, Gem Tet and so on, while the 18 pathogenic strains were sensitivity to these. PMID- 12552815 TI - [Isolation and fermentation conditions of strains producing 1-phenyl-2-amino ethanol alcohol dehydrogenase]. AB - A Arachnia sp. P163 producing alcohol dehydrogenase which is able to reduce aminoacetophenone to R-1-phenyl-2-aminoethanol was obtained from soil and cultures. The maximum activity of enzyme was produced by the LB medium containing 1% sodium citrate and peptone, 0.1% phenylaminoethanol as inducer at 30 degrees C for 48 hs. PMID- 12552816 TI - [N-terminal analysis and antibody preparation of insecticidal protein form Pseudomonas pseudoalcaligenes]. AB - The insecticidal protein from Pseudomonas pseudoaligenes was and exotoxin which had toxicity on locusts. In order to elucidate its molecular properties an amino acid sequence, the insecticidal protein was purified from the culture supernatant by ultrafiltration, ion-exchange chromatography and gel filtration, and showed a single band on SDS-PAGE. Analysis of the purified insecticidal protein dentified N-terminal sequence of ten amino acid residues. Its polyclonal antibody was also obtained by immunizing rabbit with the insecticidal protein recovered form SDS PAGE gel. The antibody titer determined by ELISA method was 1:12,800, indicating that it has high reactivity. Western blot analysis revealed that the antibody was spectific to 26 kD insecticidal protein, and did not cross-react with other proteins produced by the bacterium, suggesting that a specific antibody with high titer was obtained and could be used for further investigations of the gene cloning and expression of insecticidal protein. PMID- 12552817 TI - [Vacuolation effect of Helicobacter pylori on gastric epithelial cells]. AB - This study was designed to observe vacuolation effect of Helicobacter pylori on gastric epithelial cells. The H. pylori isolates derived from patients with peptic ulcer, chronic gastritis and gastric cancer were plated on common selective medium and their vacuolation effects on SGC-7901 cells were compared. The percentage of H. pylori(Tox+) strain in clinical isolates from different gastroduodenal diseases had no significant difference. The toxicity of the H. pylori (Tox+) strain from peptic ulcer was significantly higher than that from chronic gasritis. PMID- 12552818 TI - [Progress of biodesulfurization]. PMID- 12552819 TI - [Cloning sequencing and expression pattern, functional analysis of nifA gene in Azospirillum brasilense Yu62]. AB - The nifA gene of Azospirillum brasilense Yu62 was cloned and sequenced. The expression of nifA gene was investigated in wild type strain Azospirillum brasilense Yu62. The results show that expression of nifA gene is not repressed by ammounium and oxygen completely. But the expression of Yu62 nifA gene is different from that of strain Sp7 nifA gene. Expression of Yu62 nifA seems more sensitive to oxygen than that of Sp7 nifA which shows the highest expression in condition of aerobic, while the Yu62 nifA gene shows the highest expression in the condition of microaerobic. The regulation of NifA protein activity by ammonia and oxygen was investigated. Results showed that the NifA protein is repressed by ammonia, 1 mmol/L NH4Cl can inhibit activity of NifA protein completely. Oxygen concentration affects activity of NifA protein. NifA protein is highly active in 0.4%-0.5% O2. PMID- 12552820 TI - [Use transposon and parDE through in vivo cloning to promote the genetic stability of plasmid pCPP430]. AB - Plasmid pCPP430 carrying the hrp gene cluster after transformated into 308R (Pantoea agglomerans) can cause the hypersensitive response, and simultaneously induce the plant resistance to disease. It was genetically unstable in P. agglomerans. 0.8 kb parDE region of the broad-host range plasmid RK2 is responsible for plasmid partition. It can mediate plasmid maitenance of many kinds in Rhizobium meliloti, and also can promote the genetic stability of recombinant plasmid in the biocontrol bacteria P. agglomerans. In this paper, we cloned parDE into pCPP430 in vivo through transposition to promote its genetic stability. parDE was amplified by PCR, inserted into pGEM-T vector and cut out and religated to NotI-cut transposon vector pUT/mini-Tn5 Km to get a parDE containing mini-Tn5, pTnp. After conjugation between S17-1/lambda pir (pTnp) and 308R (pCPP430), parDE was cloned in vivo into plasmid pCPP430 to obtain pRTnp. It was demonstrated that the insertion of parDE in pCPP430 increased significantly the plasmid's stability in P. agglomerans. PMID- 12552821 TI - [Gene cloning and expression of human bactericidal/permeability-increasing protein]. AB - The gene of human bactericidal/permeability-increasing protein (BPI) was cloned from peripheral blood lymphocytes of a normal Chinese individual. The result of sequencing showed the gene is 1452 bp encoding a 27-residue signal peptide and a 456-residue matured protein, and it has six nucleotide variations compared with the sequence reported which results in 4 different amino acids. In order to get recombinant BPI, the gene was cloned into an expressing plasmid and expressed in CHO cells. The recombinant protein was purified using cation-exchange chromatography and its bioactivity was proved with bactericidal assays. PMID- 12552822 TI - [Identification and cloning of partial mbh2 gene cluster of hyperthermophile Aquifex pyrophilus]. AB - A 0.8 kb fragment of mbhS2 gene of Aquifex pyrophilus was obtained by PCR with designed primers basing mbhS2 gene of A. aeolicus. It showed 85% homology with the corresponding region of A. aeolicus. Using it as probe, a 5.0 kb Nco I fragment was fished out from the partial genomic library of A. pyrophilus. Then this fragment was cloned, subcloned and sequenced. The result revealed that the fragment contains the full length gene for the mbhS2, the gene orf1 and the first 366 bp of orf2. Comparison with mbhS2 and orf963 of A. aeolicus shows 81% and 60% homologies in amino acid sequence, respectively. PMID- 12552823 TI - [Expression of the artificial cecropin CMIV-like gene of Chinese silkworm Bombyx mori in Laphygma exigua larvae]. AB - The synthesized CMIV-like Gene was linked with the signal peptide gene of nature silkworm antibacterial peptide and was inserted into baculovirus expression vector pFastBac 1, construcing a recombinant transposing vector. The vector was transformed into DH10Bac competent E. coli cells. The recombinant Bacmid was obtained. The recombinant Bacmid was transfected into sf21 cells to get the recombinant virus. Laphygma exigua larvae were infected with the recombinant virus to express the antibacterial peptide. The hemolymph were tested to have antibacterial activity. The active antibacterial peptide was purified by acid polyacrylamide electrophoresis. The specific expression of mRNA of CMIV-like Gene was tested using Northern blotting. PMID- 12552824 TI - [Expression of the chiA gene from Serratia marcescens in both strain E26 and strain NG13, nitrogen-fixing bacteria associated with rice root]. AB - A 1.8 kb HinfI fragment carrying the chitinase gene (chiA) from Serratia marcescens was cloned into the expression vector pKK223-3 and the plasmid pMC71A, yielded the plasmid pKChiA and the plasmid pMChiA respectively. Both plasmid pKChiA and plasmid pMChiA were used to transform to the Enterobacter cloacae strain E26 and the Klebsiella oxytoca strain NG13, two nitrogen-fixing bacteria associated with rice root. The chiA gene could be highly expressed in the ChiA+ transformants of the strain E26 or the strain NG13. Cell location determination of the expressed chitinase showed that the enzyme existed not only in cell periplasm and cytoplasm, but also in extracellular broth. When the cultures were in the aftor logarithmic growthe phase, the distribution of the enzyme activity in extracellular broth, periplasm and cytoplasm were 23%-28%, 45%-51% and 21% 32%, respectively. The molecular weight of chitinase expressed in the ChiA+ transformants was 58 kD by SDS-PAGE analysis. The stability of the plasmid pMChiA in the transformants was better than that of the plasmid pKChiA. PMID- 12552825 TI - [Cloning and sequencing of 16S rRNA gene of Phytoplasma CWB1 strain associated with cactus witches' broom]. AB - A 1.5 kb DNA fragment was amplified in DNA samples extracted from Opuntia salmiana porm showed witches'-broom symptom. The result indicates the existence of phytoplasma associated with this disease and this phytoplasma was designated as CWB1. The amplified fragment was ligated to pGEM-T easy vector and then transformed into JM109 strain of E. coli. Cloned DNA fragments were verified by PCR, restriction endonuclease (EcoRI) digestion and sequence analysis. The result revealed that the 16S rRNA gene of CWB1 consists of 1489 bp and shared 99.7% homology with Faba bean phyllody which belongs to phytoplasma 16S rII-C subgroup. So we can classify this strain into phytoplasma 16S rII-C subgroup. PMID- 12552826 TI - [Melanin properties studies of the recombinant of Pseudomonas pseudoalcaligenes containing tyrosinase gene]. AB - Pseudomonas pseudoalcaligenes is a notably killing maggots bacterium, which was isolated from natural dead maggots. Tyrosinase gene of P. maltophilia AT18 has been introduced into P. pseudoalcaligenes, and enabled it the ability of producing melanin steadily. Antiradiation effect of the melanin is quite strong. The melanin of the recombinant is nonfixiform material. It's solubility is very little in many sorts of organic or inorganic solvent. It's solubility is big in alkaline solution, but little in neutral or weak acid solution. It is deposited when pH < 4. It is oxidized by H2O2 or NaC10. It is also reduced by Ag+ or H2S. It has free radical and it can absorbs free radical generated by ultra-violent ray. It can absorbs ray of all sorts of wave length. The absorption rate of ultra violet ray is the biggest in all sorts of wave length. It can effectively protects protein, DNA and other biomacromolecule matter against the damages by ultra-violet ray. PMID- 12552827 TI - [Study on the organic acids metabolism in Candida glycerolgenesis under different fermentation conditions]. AB - The organic acids in Candida glycerolgenesis fermentation medium affected both the glycerol yield and quality. When other fermentation condition was constant, the total acidity in medium increased when the volumetric ratio of medium to flask and corn steep liquid concentration were increased. When the volumetric ratio of medium to flask was 0.20 and the corn steep liquid concentration was 8 g/L, there have high yield of pyruvate and lactate produced at growth phase, 4.1 g/L and 1.0 g/L, respectively, which are two times than control sample at usual fermentation condition, hence, the glycerol yield was lower. However, the volumetric ratio of medium to flask was 0.08 and the corn steep liquid concentration was 4 g/L, the yield of pyruvate and lactate were lower and the glycerol yield was higher, but the acetate could be accumulated to 2.6 g/L. The organic acids also could be metabolized in final fermentation process, such as the initial glucose concentration was 100 g/L, after the glucose was completely utilized, both the glycerol and organic acids concentration decreased, and pyruvate or lactate could be converted to acetate in following fermentation process. In addition, the additive also can change the production of organic acids, with the addition of 1% oleic acid or VB1 the acetate concentration could be decreased and increased the production of pyruvate, hence, improved the yield of glycerol. While the fluoro-pyruvate or sulphite were added in medium, the acetate was increased largely and ketoglutarate was decreased accordingly, also increase the glycerol yields about 20%. PMID- 12552828 TI - [Production of vitamin C precursor--2-keto-L-gulonic acid from D-sorbitol by mixed culture of microorganisms]. AB - Gluconobacter oxydans SCB329 only produce a little amount of 2-Keto-L-gulonic acid(2-KLG) from D-Sorbitol when growing alone; while Gluconobacter sp. SCB110 can transform D-Sorbitol to L-Sorbose and can not produce 2-KLG. 2-Keto-L-gulonic acid, the precursor of L-Ascorbic acid (Vitamin C) synthesis, was prepared directly with a high efficiency from D-Sorbitol by mixed culture of microorganism, which comprised Gluconobacter sp. SCB110 and Gluconobacter oxydans SCB329. The fermentation product from the mixed culture broth in the D-Sorbotol containing medium was identified as 2-Keto-L-gulonic acid by HPLC, elementary analysis and infra-red adsorption spectrum. PMID- 12552829 TI - [Diversity of metabolites and their bio-activities in myxobacterium Sorangium cellulosum]. AB - Different Sorangium cellulosum strains not only showed diversity in their cell and fruiting body morphologies, but also differences of bio-activities and components of the metabolites. All the Sorangial strains studied in this paper had no activity on Gram-negative bacteria, some were able to repress Gram positive bacteria. However, all the strains were able to repress growth of fungi and tumor cells strongly and widely. Thin layer chromatography assay of metabolites showed multi-components in the metabolites, and most of them have abilities of repressing fungi and tumor cells in different degrees. Four strains were found to be able to produce compounds with activity of promoting polymerization of microtubule. Based on Rf value on TLC, the bio-active component produced by So33-1 strain was similar to Epothilone A, while that of So81 was much different. The results of this paper suggested that Sorangium cellulosum is a much beneficial resource for screening nature compounds with bio-activities against eukaryotes. PMID- 12552831 TI - [Continuous production of nisin by calcium alginate-immobilized Lactococcus lactis subsp. lacits SM526]. AB - The attempts were made to produce nisin by immobilized cells of Lactococcus lactis subsp. lactis SM526 entrapped in calcium alginate beads. The results showed that the immobilized cell beads were intact for 90 hr under 2% of calcium alginate in 10 mmol/L CaCl2 solution. The physical stability of beads was improved by removing phosphate, citrate and acetate salts supplied in SYS3 medium. Repeated-batch fermentation utilizing immobilized cells were performed in mSYS3 medium without agitation at 32 degrees C, the concentration of nisin produced during the third cycles increased to 850 IU/mL, and the beads were still intact with little cell leakage. The immobilized beads were loaded into a sterilized glass column to continuous fermentation, nisin concentration increased to 1150 IU/mL, as high as that obtained in free-cell fermentation, and the high productivity was stable for up to 70 h under a constant supplied of the fresh mSYS3 medium. PMID- 12552830 TI - [Preliminary study on citral impaires the Aspergillus flavus membrane]. AB - Compared with the normally growing A. flauas, the content as below was determined: the utilization ratio to protein and reducing sugar of hyphostroma poisened by citral, the activity of [Na+, K+]-ATPase capable of decomposition ATP, and the seepagevity ratio of electrolyte. In addition, the shape change in spore was observed via the scanning electron microscope (SEM) and the fast multi channel micro-spectrophotomer (FMCM). The result above all suggested facts as following after it's poised by the citral in MIC. The surface of hyphostroma and spore turned into be porous and rough. The pass trace on spore shriveled and closed. The rate of conduct electricity increased by 52.8%. The utilization ratio to protein and reducing sugar respectively decreased 61.5% and 44.3%. The rate of spore's sprout dropped to 61.4%. The molecular structure of membrane was so distinctly changed that it lost the selective permeability. There was inhibition on hyphostroma growth and spore sprout. PMID- 12552832 TI - [Detection of pathogenic Edwarsiella tarda]. AB - 25 Edwardsiella tarda(Et) strains had been detected both on their viruslent factor Excellular Product (ECP), including the hemolysin and extracellular protease (ECPase), and on their pathogenicity to mice and Xiphophorus helleri. ECP was detected by Dot-ELISA with rabbit antiserum against ECP of reference strain JEL4. The results showed that the animal pathogenicity of Et had good correlation with its hemolysin other than with ECPase. The agreement between Dot ELISA of JEL4 ECP and pathogenicity to animal was up to 100%. It was desirable to establish a detecting method, which only need detect the ECP with plate assay (PA) and Dot-ELISA, but needn't have animal experiment. Furthermore it is possible to develop a diagnosis kit of application to simplify the detecting procedure of pathogenic Et. PMID- 12552833 TI - [Study on the Apriona germari(Hope) larvae's intestinal bacterial flora]. AB - Intestinal flora of 47 Apriona germari(Hope) larvae, collected from fields, had been isolated and identified. The results showed that the predominant bacteria were Staphylococcus. Its viable count was 7.63 +/- 0.21, and the detection rate was 100%. Meanwhile, a strain of cellulose-utilizing bacterium was isolated from the fore-midgut fluid of A. germari larvae with the cellulose-congo red agar medium. The bacterium was tentatively identified as Cellulomonas. The detection rate of the cellulolytic bacterium was 23.40%, and the count was 3.84 +/- 0.54 approximately. Its contribution to the borer's cellulose digestion needs further investigations. PMID- 12552834 TI - [Microbial reduction ability of various iron oxides in pure culture experiment]. AB - The microbial reduction of ferrihydrite, lepidocrocite, hematite, goethite and aluminum-substituted iron oxides were examined by iron-reducer GS-15 under anaerobic pure culture condition. The results indicated that the ferrihydrite and lepidocrocite can be rapidly reduced by iron-reducer, and the percentage of microbial reduction are respectively 95.4% and 95.8% after 4 days incubation at 25 degrees C. The other iron oxides like hematite, Al-hematite, goethite and Al goethite are very difficult to reduce during short-term incubation. The amounts of Fe(III)-reducing, Fe(II)-producing and Fe-reducing calculated by acetate consumption are identical under the conditions of lower concentration iron oxides addition. PMID- 12552835 TI - [Selection of beta-glucanase-producing Trichoderma koningii T199 and its fermentation conditions]. AB - A mutant strain T199 producing about 8 times as much beta-glucanase as its parent strain trichoderma koningii T3 was obtained by treatment with ultraviolet light and N-methyl-N-nitro-N-nitrosoguanidine. Fermentation was conducted in 250 mL flask, each containing 30 mL of medium consisted of 5% corn cob powder, 3% wheat bran and 1.4% nitrogen source No. 10 ((NH4)2SO4 10%, Peptone 20%, Yeast extract 15%). The optima culture conditions were as below: initial pH 5.0, 30 degrees C, shaking speed 280 r/min, and cultivation time 5 d. Enzyme activity toward CMC-Na, lichenin, laminarin and barley beta-glucan at pH 5.0 and 60 degrees C for 10 min were 300, 1100, 12 and 1600 IU/mL, respectively. The optima pH and temperature for enzyme action toward barley beta-glucan were pH 5.0 and 70 degrees C, respectively. The enzyme was stable under below 50 degrees C and at pH 4.5-6.5. PMID- 12552836 TI - [Metabolites produced by bacteria of Xenorhabdus and Photorhabdus]. PMID- 12552837 TI - [A survey of microbial genomics projects supported by the US Department of Energy]. PMID- 12552838 TI - [Properties and mechanisms of marine polysaccharidases]. PMID- 12552839 TI - [Dual window factor analysis for data of hyphenated chromatography]. AB - A new method, dual window factor analysis (DWFA), used for the qualitative and quantitative analysis of target components hidden in chromatographic overlapping peaks of hyphenated methods is proposed in this paper. It's based on the characteristic information of the spectrum of the component. This method can directly be used for the qualitative analysis and the resolution of the spectrum and chromatogram of target components. Therefore, it is appropriate for the qualitative and quantitative analysis of the desired components in multicomponent complex system with hyphenated methods. DWFA is successfully applied to the analysis of the 4 components overlapping peak, and the results show that excellent correlations between resolved chromatographic peak area and mass concentration are kept in each of the 4 components. PMID- 12552840 TI - [Beaded molecule imprinted polymer for stereo isomer separation]. AB - Beaded molecule imprinted polymer (MIP) was made by suspension polymerization. Particles with the size of 50-70 microns in diameter were collected and evaluated in HPLC mode to separate stereo isomers. Stereo isomers cinchonine and cinchonidine were successfully discriminated with selectivity factor of 2.89 and resolution factor of 0.76. Stereo selectivity of the MIP was found to come from both the interaction between the analyte and carboxyl group on the MIP and the similarity between the stereo structure of imprinted molecule and the MIP. The thermal analysis results showed that the MIP had high thermal stability with initial thermal decomposition temperature of 320 degrees C. The pore volume of the MIP was 0.1849 mL/g, the specific surface area was 126.84 sqm/g and the average pore diameter was 5.8 nanometer. Scanning electron microscopy showed that MIP had perfect spherical morphology. PMID- 12552841 TI - [Studies on the matching relation of homologues between experimental points and parameters in retention equation in reversed-phase liquid chromatography]. AB - The matching relation between the number of experimental points and that of parameters of retention equations employing two, three, and four parameters of stoichiometric displacement model for the retention (SDM-R) of homologue in liquid chromatography has been investigated. When two experimental points were used, the retention of the homologue of N-alkylphthalimides were predicted by equations with two, three, and four parameters. The predicted values were found to fit the experimental values, although the predicted values by equation of two parameters were found to be better than that by others. Comparison was also made among the retention models proposed by Snyder, Lu Peichang, and Schoenmakers, and it is found the foregoing points have been proved. With the increase in the number of experimental points, the deviation of the predicted results of equation with multi-parameters decreased, while that by equation of two parameters did not significantly decrease. The linear correlation between the parameters of all of those equations employed in this study and the carbon number of the homologue was also discussed. The parameters from equation of two parameters was found to follow the carbon rule of a homologue. On the contrary, the parameters from the equation with multi parameters were found not to follow. PMID- 12552842 TI - [Migration behavior of low-mass anions in capillary electrophoresis]. AB - The dependence of effective mobility on ionic equivalent conductivity for 33 anions was studied using 5 mmol/L sodium chromate and 0.5 mmol/L tetradecyltrimethylammonium bromide (TTAB) as background electrolyte. The relationship was developed between effective mobility mueff and carbon number of mono-carboxylic acid and dicarboxylic acid homologues. The results showed that the effective mobility of anions increased with increasing ionic equivalent conductivity. The 1/mueff was linearly related to the carbon number of mono carboxylic acid (C2-C7) and dicarboxylic acid(C2-C10). The correlation coefficient was more than 0.997. PMID- 12552843 TI - [Hydrophobic membrane chromatography for fast purification of biological macromolecules]. AB - Cellulose membrane bonded with four commonly used hydrophobic groups, octyl, butyl, phenyl and polyethylene glycol was first investigated for their binding and purification characteristics of protein and enzyme with octyl- and phenyl Sepharose CL-4 B as controls. Hydrophobic membranes bound BSA effectively by hydrophobic interaction in high salt solution. Their binding capacities were not notably affected by significantly increasing the flow mass rate or decreasing the mass concentration of protein solution, but were much lower than those of octyl- and phenyl-Sepharose CL-4B. 11.8 fold of purification with an approximately 100% recovery of bovine liver catalase was achieved by step gradient elution on the phenyl cellulose membrane cartridge in a single step in only ten mins or a little more. Increase of the flow mass rate had no effect on the purification of catalase, however, the processing time was shortened greatly. Hydrophobic membrane chromatography here reported exibits a potential of fast processing of the protein solution in large volume with low mass concentration of the target protein, such as genetic engineering culture solution. PMID- 12552844 TI - [Chromatographic properties of tetradecylamine bonded stationary phase for reversed-phase liquid chromatography]. AB - A novel bonded stationary phase, tetradecylamine bonded stationary phase (TABP), for reversed-phase HPLC was prepared by bonding 1-tetradecylamine to YWG-80 silica gel through 3-glycidoxypropyltrimethoxysilane. Hydrophobicity, selectivity and silanophilic activity of TABP were evaluated by using aromatic compounds as analytes and methanol-water as binary mobile phase. The orgainc components including acidic, basic and neutral aromatic analytes could be separated satisfactorily with excellent selectivity and peak shape. The relative retention alpha for ethylbenzene-toluene was found to be 1.66 and the asymmetry factors of basic aniline, p-toluidine and N,N-dimethylaniline were found to be 1.26, 1.21 and 1.10, respectively, with the V(methanol):V(water) = 55:45 mobile phase. Aniline was eluted before phenol due to the suppress of the ion exchange activity of residual silanols by the internal masking interaction. PMID- 12552846 TI - [Capillary electrophoresis of polysaccharides and its application]. AB - This paper deals with the progress in capillary electrophoresis of migration, detection and its applications on polysaccharides. The papers of capillary electrophoresis of polysaccharides in the last 20 years are reviewed. PMID- 12552845 TI - [Theory and application of affinity capillary electrophoresis]. AB - Affinity capillary electrophoresis (ACE) is a technique for studying receptor ligand interactions. ACE experiments are based on the different values of electrophoretic mobility of free and bound receptor. ACE can be introduced to many capillary electrophoresis models, such as CZE, MECC, CIEF, CGE etc. It's applications include immunoassays, determining the binding constants of formed complexes, screening libraries for lead compounds, sequence-specific and base composition-specific recognition of oligodeoxy-nucleotide etc. PMID- 12552847 TI - [The application quantitative structure-retention relationship of GC to aid MS qualitative analysis]. AB - In this paper, a method, based on the prediction of the Kovats retention indices of related polysubstituted alkylbenzenes from the indices of benzene, monsubstituted alkylbenzenes and disubstituted alkylbenzenes, was used to aid qualitative analysis with mass spectrometry alone. A disproportinated aromatic product and the basic compounds of alkylbenzene were analyzed with gas chromatography isothermally at 100 degrees C separately. In order to cut down the analysis time and increase the resolution, the disproportionated aromatic product was analyzed using temperature programming with gas chromatographymass spectrometry (GC-MS). The Kovats retention indices of the basic compounds were determined and those of the polysubstituted alkylbenzenes were predicted with the method which was proposed by us previously. The isomers of polysubstituted alkylbenzenes have similar mass spectra, so they can not be identified unambiguously by MS alone. For the differences of the Kovats retention index between isomers of polysubstituted alkylbenzenes are larger, it can be identified by matching the experimental retention indices with the predicted ones. The positional isomers of polysubstituted alkylbenzenes which are very difficult to be interpreted by MS alone have been identified explicitly. PMID- 12552848 TI - [Gas chromatographic method for the analysis of polychlorinated biphenyls in pine needles]. AB - Polychlorinated biphenyls (PCBs) are a class of 209 chemical compounds, in which 1-10 chlorine atoms are attached to a biphenyl molecule. PCBs are members of halogenated aromatic group of environmental pollutants that have been identified worldwide in diverse environmental matrices. PCBs in air, soils, sediment, water, transformer oils and other environmental matrices have been determined in the past years. In this work a method for routine analysis of PCBs in pine needles has been developed. First, extractions were carried out in Soxhlet apparatus with n-hexane as solvent. Then, a silica gel chromatographic column was applied to pretreat the pine needle samples. The reference standard used was clophen 50. The recovery was about 90%, so the feasibility and reliability were assured. After the analysis of PCBs in pine needle samples from four different regions, the distribution of PCBs in environment can be discussed and monitored. In this method, a Shimadzu GC-7A gas chromatograph equipped with a 63Ni electron capture detector was used for the analysis of PCBs. From the results, we can see the PCB pollution in different regions. So evaluation of air pollution level through foliage data is feasible. The sample preparation and analytical method mentioned in this paper is reliable and simple. PMID- 12552850 TI - [Study on the analytical method of methyl-parathion and phoxim in methyl parathion, phoxim and cypermethrin mixed formulation]. AB - This paper describes an HPLC method to simultaneously determine methyl-parathion and phoxim in mixed formulation of methyl-parathion, phoxim and cypermethrin. A C8 column was used. The mobile phase was V(MeOH):V(H2O) = 70:30, Methyl-parathion and phoxim were monitored at 254 nm or 280 nm by UV detector. The average recoveries were 99.8% and 99.9% for methyl-parathion and phoxim respectively. The linearity of detector response vs concentration was calibrated in the concentration ranges from 0.1746 to 1.091 g/L for methyl-parathion and from 0.1927 to 1.205 g/L for phoxim. PMID- 12552849 TI - [The resolution of racemic sec-phenethyl alcohol on cellulose tribenzoate-based CSP: influence of different alcohols in the mobile phase]. AB - Several primary and secondary alcohols (ethanol, 1-propanol, 2-propanol, 1 butanol) were used as the mobile phase components separately, to investigate their effects on the capacity factor and stereoselectivity of sec-phenethyl alcohol enantiomers on cellulose tribenzoate-based CSP. The chiral recognition mechanism for the enantiomeric aromatic alcohols studied may involve: (1) the aromatic portion of the solute may insert into a chiral cavity of the CSP through a hydrogen bonding interaction between the solute's alcoholic hydrogen and the ester carbonyl group on the CSP; (2) the mobile phase modifiers (various alcohols) compete with the solutes for chiral, as well as achiral, binding sites on the CSP; (3) the structure of the modifier has some effect on stereoselectivity through an alteration of the steric environment of the chiral cavity. PMID- 12552851 TI - [Chiral separation of thioglycidyl ether and glycidylselenide on cyclodextrin bonded stationary phase via high performance liquid chromatography]. AB - beta- Cyclodextrin bonded chiral stationary phases (CD-CSP) were prepared after the method of Armstrong (CD-CSP I) and of Huang Tianbao (CD-CSP II). With the separation of o-, m- p-nitroaniline, it was found that the chromatographic properties of CD-CSP II was superior to that of CD-CSP I. Three racemes of thioglycidyl ethers and two racemes of glycidylselenides were resolved on CD-CSP II via high performance liquid chromatography. The mobile phase consisted of different volume ratios of methanol/0.3% triethylammonium acetate buffer solution(pH 5), and the eluate was detected with a UV-visible detector at a wavelength of 254 nm. With the increase of methanol volume ratio in the mobile phase, the retention time decreased rapidly, but within the range of methanol valume ratio 40/100 to 50/100, the separation factor only decreased slightly. The racemes examined contained aromatic groups and hydroxy groups which can form inclusion complexes and hydrogen bonding interactions with beta-cyclodextrin. The chiral separation of the racemes is successful with the mobile phase of V(methanol):V(buffer) = 40:60. PMID- 12552852 TI - [Determination of resveratrol in grape wine by high performance liquid chromatography]. AB - Resveratrol is a microactive factor for preventing human cancer. A reversed phase high performance liquid chromatographic method for the determination of resveratrol in grape wine was developed. Samples were extrated by ethyl acetate and washed with 3% NaHCO3; after dehydrated with Na2SO4 and dried in vacunm at 40 degrees C, dissovled in acetonitrile. The conditions of HPLC analysis were as follows: the maximum absorptions of trans-resveratrol and cis-resveratrol were at 306 nm and 280 nm respectively. Separation was achieved by the use of mu-Bondapak C18 column (3.9 mm x 300 mm). The mobile phase was V(acetonitrile):V(water) = 40:60. The retention times of trans- and cis-resveratrol were 7.7 min and 8.9 min respectively. The recovery rates were 90.0%-97.8% and the CV was 3.3%. Detection limit was 10 micrograms/L. Eleven kinds of red and white grape wine made in China were detected. All of them contained resveratrol, but the quantity, from 10.0 to 920.0 micrograms/L, were quite different. PMID- 12552853 TI - [Separation of enantiomeric labetalol by reversed-phase high performance liquid chromatography]. AB - A reversed-phase high performance liquid chromatographic method for the separation of labetalol enantiomers was developed. In the method, 2, 3, 4, 6 tetra-O-acetyl-beta-D-glucopyranosyl isothiocyanate (GITC) was chosen as the reagent for pre-column chiral derivatization of labetalol to give four diastereomeric thiourea derivatives. These derivatives were efficiently separated on a Nova-Pak C18 column using V(MeOH):V(0.01 mol.L-1(pH 7.00) H2PO4(-)-HPO(4)2- buffer) = 51:49 as the mobile phase and detected by UV detector at a wavelength of 250 nm or fluorescence detector at lambda ex = 340 nm and lambda em = 440 nm. The effects of the pH of mobile phase on the retention and fluorescence absorbance are also discussed. PMID- 12552854 TI - [Instrumental analysis and systematic investigation on heavy oils from coal. I. Analysis of hydrocarbon class composition]. AB - A study on the analysis of hydrocarbon class composition was carried out by thin layer chromatography with flame ionization detection(TLC/FID). The optimum analytical conditions, experimental reproduction, standard deviation, quantitative correction factors of hydrocarbon class and so on were investigated. The instrument used were Mark III, LATROSCAN TH-10 analyzer, thin layer rod was a CHROMAROD silica rod(150 mm x 0.9 mm i.d). A satisfactory TLC/FID analytical method was established after a series of experiments and investigation. Typical heavy oils from coal were investigated based on this method. The optimum conditions were that the flame ionization detector was operated at a hydrogen flow rate of 100 mL/min corresponding to 90 kPa and air flow rate of 1.9 L/min. The determinations in groups of saturated hydrocarbons, aromatics, resins and asphaltenes were performed by using n-heptane, toluene and V(chloroform):V(methanol) = 95:5 as developing solvents. The developing distances were controlled at 10 cm, 5.5 cm and 2.5 cm respectively. In the present paper qualitative and quantitative analysis have been studied. And the method established was sufficiently applied to determine heavy oils from coal of coking plants of Shanxi. PMID- 12552855 TI - [Determination of anions in recycle sodium formate from sodium hydrosulfite industry with single column ion chromatography]. AB - The analysis of recycle materials from industrial waste water of sodium hydrosulfite production was made by single-column anion exchange chromatography. The recycle material was sodium formate (HCOONa) in which various anions, such as SO(3)2-, Cl-, thiosulfate, hydroxyethyl sulfonate (HOC2H4SO3-), hydroxyethyl thiosulfate (HOC2H4S2O3-), SO(4)2- and NO3- as impurities were existed simultaneously. Except hydroxyethyl sulfonate anion, all other ions can be separated by using anion exchange column Shim-pack IC-Al (4.6 mm i.d. x 100 mm) with the mixture of 1.8 mmol/L of phthalic acid and 1.35 mmol/L of Tri(hydroxymethyl) aminomethane as eluent. The flow rate of mobile phase is 1.0 mL/min. The injection volume was 20 microL. The column oven temperature was controlled at 40 degrees C. Hydroxyethyl sulfonate anion and SO(3)2- could not be separated at the above conditions, but they did not interfere the determination of HCOO-. The detection limits for HCOO-, SO(3)2-, NO3-, S2O(3)2-, HOC2H4S2O3-, Cl- and SO(4)2- were 1.0, 0.7, 1.4, 5.0, 0.7, 0.2 and 2.0 mg/L, respectively. The recoveries were between 96%-102%, and the relative standard deviations (RSD) were lower than 6.0% (n = 5) for all seven ions. This method is characterized by rapidity, sensitivity and simultaneous determination of several kinds of ions. PMID- 12552856 TI - [Influence of sample dilution on separation and detection of DNA fragments by capillary electrophoresis]. AB - Capillary electrophoresis has become an important and useful method to separate and determine DNA fragments. In molecular biochemistry, the volume of DNA sample is very small (microL level) and DNA sample is liable to be contaminated and degraded. According to theoretical inference and experiments, we propose that dilution of DNA sample solution can increase separation efficiency and resolution without evidently reducing height of peaks. By this method, the usage efficiency of DNA sample can be improved. It is also demonstrated the separation and detection of DNA fragments by capillary electrophoresis with hydroxyethyl cellulose non-gel sieving matrix and with laser-induced fluorescence charge coupled device as detector. By using lower concentration non-gel matrix (0.4%), all 8 larger size fragments of lambda DNA/Hind III (125 bp-23 130 bp) can be completely separated. Twenty smaller size fragments of pBR322-Hae III DNA (18 bp 587 bp) can be separated by higher concentration (1.6%) non-gel matrix. As ratio of sample dilution is 10, two adjacent fragment (123 bp and 124 bp) of pBR322-Hae III DNA can be separated. PMID- 12552857 TI - [Separation and analysis of amino acids in secretion of psylla chinesis yang et li]. AB - We used ion-exchange resin column chromatography to separate amino acids in secretion of Psylla Chinese Yang et Li and used capillary gas chromatography to analysis n-butyl-N-trifluoracetyl derivatives of the amino acids. Qualitative analysis was done by comparing with the standard amino acids and quantitative analysis by internal standard method. We determined thirteen amino acids in the secretion. PMID- 12552858 TI - [Determination of pentachlorophenol residues in textiles and leather and leather products by gas chromatography/mass spectrometry]. AB - A method for determination of pentachlorophenol residues in textiles and leather and leather products by gas chromatography/mass spectrometry was developed. The pentachlorophenol residues in samples was acidified with sulphuric acid solution (6 mol/L) and extracted with n-hexane. After the n-hexane layer was washed with sodium sulphate solution (20 g/L)until it was clear, determination was made by means of a gas chromatograph equipped with MSD, using external standard method. A DB-17 fused silica capillary column (30 m x 0.25 mm i.d.; 0.25 micron film thickness) was employed in the analysis, The column temperature program included a 2 min isothermal period at 50 degrees C, temperature increased at a rate of 30 degrees C/min to 220 degrees C, and hold for 1 min; temperature increased of at a rate 6 degrees C/min to 260 degrees C, and hold for 1 min. Mass spectra were obtained by electron impact at 70 eV and the determination was made by means of SIM mode, selected monitoring ion (m/z) was 264 amu. The detection limit was 0.02 mg/kg. The average recovery was from 86.7% to 93.1% and CV (n = 10) is from 4.1% to 5.9%. This method was already used to determine the mass concentration of pentachlorophenol residues in raw cotton, towel, shirt, leather gloves, leather watchguard. The method is simple, rapid and accurate. PMID- 12552859 TI - [Determination of C1-C5 composition of liquefied petroleum gas by capillary gas chromatography]. AB - In this paper, thirteen C1-C5 hydrocarbons in liquefied petroleum gas were separated and determined by using capillary gas chromatography. The cloumn used was 50 m x 0.53 mm i.d. HP PLOT-Al2O3. A standard mixture of known composition was used for the determination of correction factots and the corrected area mormalization was used for quantification. The relative standard deviations (n = 5) were 0.39%-5.4%. Ethane, ethylene and butene were separated to baseline. PMID- 12552860 TI - [Determination of itaconic acid in culture broth by high performance liquid chromatography]. AB - An HPLC method for the determination of itaconic acid and other components in the culture broth of Aspergillus terreus was developed. The operating conditions were Aminex HPX-87H column (300 mm x 7.8 mm) at 30 degrees C, mobile phase of sulfuric acid (4 mmol/L) and refractive index detector at 30 degrees C. The linear relationship of calibration curve was good in the range of 0-50 g/L for itaconic acid (r = 0.9997). The detection limit was 5.7 micrograms, and the relative standard deviation was 1.37% (n = 7). Other components in the culture broth were SO(4)2-, sucrose, glucose and fructose. The method is simple, sensitive, rapid and accurate. PMID- 12552861 TI - [Determination of sarafloxacin by high performance liquid chromatography (HPLC)]. AB - A method for the separation and determination of Sarafloxacin by HPLC was developed. The sample was dissolved in a mixture of V(acetonitrile):V(water) = 1:1. Operating conditions were as follows: mu-Bondapak C18 column (3.9 mm x 300 mm), V(acetonitrile):V(methanol):V(2 mmol/L H3PO4, adjust pH 3.5 with triethylamine) = 30:5:65 as mobile phase with a flow rate of 1 mL/min, UV detection at 278 nm, and column temperature was 15 degrees C. Under the above conditions, Sarafloxacin and other impurities were separated from each other. The method was simple, rapid, sensitive and accurate. PMID- 12552862 TI - [Determination of risperidone and 9-hydroxyrisperidone in plasma by high performance liquid chromatography]. AB - A reversed-phase HPLC method for the determination of risperidone and 9 hydroxyrisperidone in plasma was established. Risperidone and 9 hydroxyrisperidone can be separated on a Zorbax ODS C18 column with a mobile phase of MeOH-water-ammonium acetate (mol/L)-ammonia water(3 mol/L) (300:50:3:1, volume ratios) and detected at 280 nm. The flow rate was 0.8 mL/min. The linear range was 2-600 micrograms/L, r = 0.996 for risperidone, and 2-800 micrograms/L, r = 0.998 for 9-hydroxyrisperidone. The average recovery was (98.2 +/- 3.5)% for risperidone, and (97.8 +/- 3.8)% for 9-hydorxyrisperidone. The intra-day and inter-day coefficients of variation were 4.12% and 4.83% for risperidone respectively, 4.28% and 4.81% for 9-hydroxyrisperidone respectively. The method has been used to determine risperidone and 9-hydroxyrisperidone mass concentration in patient plasma. The data obtained showed that the sensitivity and selectivity of this method were adequate for drug monitoring in clinical research. PMID- 12552863 TI - [Analysis of HPLC of isomers of nitrotoluene and nitrobenzoic acid]. AB - Nitrotoluene and nitrobenzoic acid isomers were separated completely in less than 10 minutes by Kromasil C18 (200 mm x 4.6 mm i.d.), when V(CH3OH):V(H2O):V(THF) = 55:44:1 with beta-CD of 0.02 mol/L at flow rate of 2.0 mL/min (former 4 min) and 2.6 mL/min (later 6 min). Detector of UV-254 at 0.04 AUFS was used. PMID- 12552864 TI - [Separation and determination of morphine, pethidine and diazepam by high performance liquid chromatography]. AB - Morphine, penthidine and diazepam in blood were determined by high performance liquid chromatography (HPLC). The HPLC system was consisted of 10 microns particle size C18 column and UV detector set at 254 nm. The mobile phase was V(CH3OH):V(25 mmol/L KH2PO4) = 90:10 with a flow rate of 1.0 mL/min. The average recoveries of morphine, penthidine and diazepam were 93.27%, 90.23% and 93.29% respectively. The linear range was between 0.05-50 mg/L. The detection limit was 0.05 mg/L and CV was less than 6%. PMID- 12552865 TI - [Determination of pericarpium papaveris(yingsuqiao) in bottom material of chafing dish by high performance liquid chromatography(HPLC)]. AB - Morphine, codeine and papaverine in bottom material of chafing dish were determined by HPLC. The conditions were as follows: C18 column, mobile phase V(CH3OH):V(25 mmol/L KH2PO4) = 90:10, the flow rate 1 mL/min and the wavelength 254 nm. These three compounds were entirely separated. The blank and positive comtrasts were being done simultaneously. The detection limit was 0.1 mg/L. The experiment showed that this method is sensiuve, accurate and reliable. PMID- 12552866 TI - [Determination of enantiomeric purity for epinephrine by high performance liquid chromatography]. AB - A procedure for the resolution of D,L-epinephrine and the determination of D isomer ratio in a pharmaceutical formulation is described. The optical isomers of epinephrine were resolved by RP-HPLC with heptakis(2,6-di-O-methyl-beta cyclodextrin) as a chiral mobile phase additives. The results showed that a correction factor must be introduced into the equation used for the calculation of the percentage of D-epinephrine, because the UV absorbance may not be the same when DM-beta-CD is present. Quantitation was achieved with external and internal standard method, HVA as an internal standard. This method can be used for the determination of the D-isomer in the L-epinephrine pharmaceutical formulation. The results showed that the mass fraction of D-isomer in the formulation increased during the period of storage. PMID- 12552867 TI - [Determination of conversion of asymmetric hydrosilylation of acetophenone by gas chromatography]. AB - A gas chromatographic method to determine the conversion of asymmetric hydrosilylation of acetophenone has been established. The gas chromatographic conditions were as follows: column, 10% PEG20M on white 102 support (DMCS), 1 m x 2 mm i.d. stainless-steel column; column temp., 160 degrees C; injector temp., 200 degrees C; thermal conductivity detector temp., 190 degrees C; carrier gas H2 with flow rate of 50 mL/min. The results show that the method is simple and rapid, and has good reproducibility. The coefficient of variation of the method was less than 1% (n = 6). The results were also confirmed with 1HNMR. PMID- 12552868 TI - [Rapid determination of saccharin sodium in ice cream by high performance liquid chromatography (HPLC)]. AB - This is a method for the rapid determination of saccharin sodium in ice cream by HPLC. A radial compression C18 cohumn, 100 mm x 8 mm i.d., and ultraviolet detector at 220 nm were applied to the determination without extraction. The solution of samples would be determined after they were treated with the solution of copper sulfate and sodium hydroxide in water. The detectable amount was 0.38 mg/kg, the coefficient of variation CV was 1.72%, and the recovery was 96.25% 101.75%. PMID- 12552869 TI - [Determination of the azetidine-2-carboxylic acid from lily of the valley]. AB - This paper reports the separation and determination of the azetidine-2-carboxylic acid in lily of the valley by high performance liquid chromatography (HPLC) with post-column derivatization and fluorescence detection. The operating conditions were cation exchange resin column (30 cm x 0.4 cm i.d.) with buffer solution A [1.96% (mass fraction) sodium citrate solution pH 3.10 +/- 0.02] and B [2.1% (mass fraction) sodium nitrate solution pH 9.60 +/- 0.02] as mobile phase at flow rate of 0.4 mL/min, gradient elution, column temperature 65 degrees C, detection wavelength lambda ex = 338 nm and lambda em = 425 nm. The retention time of azetidine-2-carboxylic acid was 5.86 minutes. The recovery was 96.4% and coefficient of variation was 0.94%. This method is simple, rapid and sensitive. PMID- 12552870 TI - [Separation of dicarboxylic acid by thin-layer chromatography]. AB - Dicarboxylic acids (C2-C6) were detected on thin-layer chromatography of silica gel G after developing with V(benzene):V(acetic acid):V(water) = 50:40:10 for an hour by dipping the plate in a very short time. The chromogenic reagent was 0.1% bromocresol green alcohol solution adjusted to pH 7. The acid appeared as yellow spots on a blue background. PMID- 12552871 TI - [Analysis of mathematical model parameters for continuous rotary annular chromatography]. AB - The curves of analysis solution of mathematical model for continuous rotary annular chromatography (CAC) coincide with experimental results well, but some points are derived from theoretical values due to the errors from experimental method. The width and the angles of maximum mass concentration solution at outlet (retention number), except for the other experimental conditions, depend on phase equilibrium coefficient and transfer functions. Because the precision of model parameters is strongly affected by the correction of experimental method, the sensitivities of phase equilibrium coefficients and transfer functions have been studied, in which the phase equilibrium coefficient is the key parameters to improve the separation effect of CAC process. PMID- 12552872 TI - [Analysis of products of thymine irradiated by heavy ion beam with capillary gas chromatography]. AB - In this study some chromatographic methods with capillary column, such as GC, GC MS, GC-FTIR, were used to analyze the products of thymine irradiated by 18O8+ ion beam in N2O saturated aqueous solution. Because of the complexity and small amounts of the products after irradiation by heavy ion beams, it is very difficult to separate them, while with the methods of capillary gas chromatography this problem can be overcome. From the results of GC-MS analysis we can determine the molecular weight of the product, and from the results of GC FTIR analysis we can get structure information of the product. By this way the products such as 5,6-dihydrothymine, 5-hydroxyl-5-methylhydantoin, 5-hydroxyl-6 hydrothymine, 5-hydro-6-hydroxylthymine, 5-hydroxymethyluracil, trans-thymine glycol, cis-thymine glycol and dimers were determined. Though these products are the same as those from thymine irradiated by gamma ray in N2O saturated aqueous solution, the mechanism of heavy ion beam interact ion with thymine aqueous solution is different from that of gamma ray. The main products from thymine irradiated by 18O8+ ion beam in N2O saturated aqueous solution are formed by hydroxyl additions between 5-6 band of thymine, while those by gamma ray are dimers of thymine. PMID- 12552873 TI - [Estimation of the octanol-water partition coefficients of PAHs by solid-phase microextraction]. AB - Eleven PAHs were analyzed by solid-phase microextraction-gas chromatography/ion trap detector and their partition coefficients were obtained at equilibrium. When the polymer coating of the fiber may be viewed as one kind of organic solvent, the linear free energy relationship between the polydimethylsiloxane-water partition coefficient (Ksw) and octanol-water partition coefficient (Kow) was established by solid-phase microextraction, which was log Ksw = 0.9318 log Kow 0.2056 with good correlation coefficient 0.9504. The linear equation may be used to estimate the octanol-water partition coefficient of other PAHs and similar H acceptors compounds. Being compared with Leo's method which was based on the additive-constitutive nature of the partition coefficient, the solid-phase microextraction method may distinguish efficiently Kow of the isomer. PMID- 12552874 TI - [Cibacron blue F3GA-attached 2 microns non-porous monodisperse silicas for affinity chromatography]. AB - Non-porous monodisperse silica (NPS), 2 microns in diameter, was modified with 3 aminopropyltriethoxysilane for immobilization of Cibacron Blue F3GA (CB), a packing of NPS-ACB for affinity chromatography was obtained. Up to 2 mg of CB could be attached to 1 mL of NPS beads. There was no obvious leakage of dye from NPS-ACB. Oval was not retained by the column, while Lys was specifically adsorbed. The adsorption of Lys varied with pH values and ionic strengths. In addition, alpha-globulin could not be retained by the packing, while beta- and gamma-globulin could be adsorbed on the column. gamma-Globulin was able to be eluted by 20% 1,6-hexanediol and 1 mol/L KCl, while beta-globulin was not able to be eluted by the same eluent. The difference in affinity interaction could be used to separate the three globulins. Furthermore, the column could be used for separation and preparation of Lys from hen egg white. The chromatograms of Lys on non-porous silica diethylamine column (NPS-DEA) showed that retention time of one peak of the crude Lys prepared was in accordance with Lys's, so it could be said that NPS-ACB column can be used for preparation in a small scale. PMID- 12552875 TI - [Preparation and evaluation of new ion-exchange chromatographic stationary phase for the use in high performance liquid chromatography]. AB - A new method for the bonding of diethylamine(DEA) on the surface of silica to prepare novel hydrophilic packings for HPLC has been studied. After allyl glycidyl ether being synthesized, the Si-DEA anion-exchange bonded phase was prepared by the reaction of the double bond in allyl group with Si-H silica. The bonded phases obtained were characterized by elemental analysis, diffuse reflectance infrared Fourier transform(DRIFT) spectroscopy and HPLC evaluation. The methods were used for both porous silica and monodisperse non-porous silica. The contents of carbon, hydrogen and nitrogen of porous Si-DEA packing (MPS-DEA) were 3.31%, 0.95% and 1.34% respectively and those of monodisperse non-porous Si DEA packing (NPS-DEA) were 2.55%, 0.97% and 0.96% respectively. The diethylamine absorption peak can be observed at 2970 cm-1 from the Si-DEA silica DRIFT spectrum. These data revealed that the diethylamine had been bonded on MPS-DEA and NPS-DEA packings. In HPLC tests, nucleotides and nucleosides such as cytosine, uracil, cytidine-5'-monophosphate, adenosine-5'-monophosphate, inosine 5'-monophosphate and guanosine-5'-monophosphate were satisfactorily separated on the porous anion-exchange packing (MPS-DEA), and a group of proteins (lysozyme, ribonuclease, ovalbumin, bovine serum albumin, insulin and gamma-globulin) were separated within 15 minutes successfuly. All test results indicated that the new method for preparing better anion-exchange silica packings is effective for both porous silica and monodiperse non-porous silica. PMID- 12552876 TI - [Determination of epinephrine, chlorpromazine and promethazine by capillary electrophoresis with scanning voltammetric detector]. AB - With a self-constructed electrochemical detection system, the determination of epinephrine, chlorpromazine and promethazine has been studied by using capillary electrophoresis with scanning voltammetric detector. Scanning potential generation and data acquisition were carried out with D/A and A/D converter board that mounted on a 486DX66 computer. A complex software, which was written by authors in BORLAND C++, was serve to data acquisition, abstraction of three dimension data, display and print of the electrophorograms. Using a 25 microns x 70 cm fused-silica capillary, the optimum conditions for separation and detection of titled drugs are 10 mmol/L K2HPO4 + H3PO4 (pH 3.0) running electrolyte, 20 kV separation voltage, hydrodynamic injection 10 s at 20 cm height, scanning potential from +0.4 to +1.0 V vs. SCE. With the optimizing condition, the separation of titled drugs is satisfactory, and good reproducibility and low limit are obtained. In this study, all experiments are carried out under air conditioning (18 degrees C) and constant humidity (relative humidity 50%). In comparison to amperometric detection, scanning voltammetric detection has the advantages of minimizing fouling of the working electrode surface, and direct identification of unknown chemicals by voltammetric curves. PMID- 12552877 TI - [Separation and characterization of oligodeoxynucleotide by fast protein liquid chromatography]. AB - The antisense drug synthesized by DNA synthesizer is an oligodeoxynucleotide (ODN). The purity of the drug should be determined effectively. Because the ODNs are molecules with negative charges at pH 12, they can be separated according to the quantity of charges with them on the ion-exchange column. On anion exchange column, MONO-Q, the ODNs ranged from 19 bases to 21 bases can be separated successfully by gradient elution of NaCl solution from 0 mol/L to 1.8 mol/L. The experiment proved that the fast protein liquid chromatography is a very useful tool for the determination of the antisense drug. PMID- 12552878 TI - [The advances of the study on the optical chromatography]. AB - This review is mainly concerned with the recent developments in (1) the differences between the optical tweezer and optical chromatography; (2) the principle of optical chromatography separation; (3) the theory studies on the optical chromatography, especially the relationship between the particle radius with the intensity of the applied laser radiation force and the relationship between the particle retention distance with the particle radius; (4) the application of the optical chromatography technique to the separation of polymers, biological macromolecules and biological cells. The application of optical chromatography to the immunoassy and molecular biology is highlighted. PMID- 12552879 TI - [Liquid chromatography with amperometric detection]. AB - Amperometric detection in liquid chromatography is introduced in this article. Detection mechanism and conditions involved in the application in HPLC and HPIC with d.c. amperometry, pulsed amperometry and integrated amperometry detection are discussed. PMID- 12552880 TI - [Prediction and estimation on molar response values of alkane using molecular subgraph]. AB - A new method based on topographical subgraph of alkanes is proposed and used for molar response values of alkanes. A novel coding, which derived from the numbers of molecular subgraph, is used to characterize molecular structure of alkanes. It showed that there is a very good correlation between the coding and molar response values. The results of multiple linear regression are as follows. Molar response values (SM) on FID: SM = 89.579 - 56.579x1 - 40.579x2 - 18.191x 3 - 2.677x4 + 11.818x5 + 25.690x6 + 40.421x7 + 54.421x8 + 13.926x9 + 15.524x10 + 13.923x11 + 13.857x12 + 30.079x13 + 30.403x14 + 32.677x15 + 29.030x16, N = 50, r = 0.9983, S = 1.9342; Molar response values (SM) on TCD: SM = 139.799 - 62.251x2 49.214x3 - 31.244x4 - 15.992x5 + 0x6 + 13.201x7 + 27.201x8 + 11.152x9 + 9.225x10 + 8.427x11 + 10.041x12 + 22.452x13 + 14.977x14 + 22.318x16, N = 32, r = 0.9963, S = 1.7668. Where r, S and N are regression coefficient, residual standard deviation and data number, respectively. Having tested the modeling by using back propagation neural network (BPNN) and leave-one-out method, we got the correlation coefficient of cross validation, 0.989. PMID- 12552881 TI - [The study of carbonaceous adsorbent for solid-phase microextraction and the comparison with commercial device]. AB - The parameters governing solid-phase microextraction have been investigated, and the development of carbonaceous adsorbent for SPME is presented because it is durable and inexpensive. The time profile curves were determined for several organic pesticides having a wide range of water solubilities and the effect of salt was also investigated. The method of SPME-GC-ECD to analyse trace level of the organic pesticide in water is established by the determination of optimum parameters. The comparison with commercial SPME device shows that the carbonaceous adsorbent having been treated with physical or chemical method for SPME can get lower limits of detection than polyacrylate and polydimethylsiloxane, although there are still some work to be done to improve its precision. The results shows that this new SPME device has great application potential. PMID- 12552882 TI - [Preparation technique of S2OF10 gas standard sample and determination method of the trace S2OF10 in SF6]. AB - In this paper a series of methods and techniques for the S2OF10 standard sample preparation and quantitative determination are presented. They are, the preparation of S2OF10 by preparative chromatography with adsorption/thermal desorption, the standard sample of S2OF10 gas prepared by exponential dilution and the gas chromatography/flame photometric detector(GC/FPD) determination of trace S2OF10 from an SF6 sample with quantitative calibration factor. Especially, the S2OF10 gas from a used SF6 sample was directly separated and concentrated through a U-adsorbent-tube packed with 300 mg of Porasil A in a cold trap (-63 degrees C) with liquid-nitrogen and chloroform. Then it was purified by preparative-GC and to be injected into a preparative system of standard gas sample. In the meantime, the S2OF10 gas obtained was confirmed by the methods of GC/FPD, infrared spectrophotometer(IR) and gas chromatography/mass spectrometer(GC/MS) separately. The sub-ppm(by volume) level of the S2OF10 and SF6 mixture samples were prepared by use of the exponential dilution system. The GC/FPD experimental results showed that the detection linear range of S2OF10 gas concentration was 0.80 x 10(-6)-2.60 x 10(-4) (volume fraction) and the quantitative calibration factor of the S2OF10 was 0.197 based on SF6. The determination errors of quantitative calibration factor were 1.8%-20% and S2OF10 recovery of the adsorption/thermal desorption was 98.2% (n = 9) and its relative standard deviation was 6.2%. In addition, the results also showed that it is a simple and rapid method with good linearity and reproducibility. PMID- 12552883 TI - [Direct determination of purine bases in tea by reversed-phase high performance liquid chromatography]. AB - A reversed-phase high performance liquid chromatographic(RP-HPLC) method for the direct determination of three purine bases(theobromin, theophyllin and caffeine) in tea was developed. An ODS column with Zorbax SB-C18(4.6 mm i.d. x 250 mm, 5 microns) was employed. The aqueous solution of methanol containing 0.05% of acetic acid and 0.25% of N,N-dimethylformamide(DMF) was used as eluent with a flow rate of 0.8 mL/min. In this method, the aqueous extract of tea can be injected into HPLC directly, but in current HPLC methods for purine bases the coexisted tea polyphenols must be pre-separated. The three purine bases in tea were separated without any interference from the coexisted tea polyphenols. This method is simple (without any special sample pretreatment) and sensitive with detection limits (S/N = 3) of 0.7, 0.9 and 1.8 mg/L for theobromin, theophyllin and caffeine respectively. The linear range of the calibration curve of peak area for the three purine bases were from 6 mg/L to 1,000 mg/L with a correlation coefficient (r) of 0.998-0.999. PMID- 12552884 TI - [Correlation of conformational change with enzyme activity of glucose isomerase in denaturants monitored with high performance liquid chromatography]. AB - Glucose isomerase (GI) can catalyze in vitro the isomerization of D-glucose to D fructose. So it is an extremely important industrial enzyme in the commercial conversion of starch to high fructose syrups. In the previous papers, we have purified and characterized the enzyme from streptomyces diastaticus M1033 of China and obtained the crystal structures by X-ray. In this paper, a method for measurement of the dynamic conformational change procedure of glucose isomerase in various concentrations of denaturants by HPLC has been established. At first the relative molecular mass of GI in solution is measured by HPLC on PROTEIN PAK 300SW (7.5 mm i.d. x 30 cm) column. The relative molecular mass of GI is about 150,000. So GI exists as tetramer in the solution without denaturants. In 0-5 mol/L guanidine hydrochloride, incubated at 30 degrees C for 30 min, GI is gradually dissociated into monomer, and at the same time its activity gradually disappears. In various concentrations of urea and incubation at 30 degrees C for 30 min (or 60 degrees C for 1 h), the results are different from that in guanidine, because the monomers peaks of GI is not found. Only in certain concentrations of urea, the small dimer peaks of GI is found, but the activity of GI significantly disappears. Moreover as the increase of the urea concentration, the retention time of tetramer peak is gradually decreases. From the fluorescence spectra, we found the conformation of GI changed in the solution of urea. So perhaps in urea, the conformation of GI become a little unfolded, and the active region is partly damaged, which makes GI partly inactive. Dissociation into inactive monomers and conformation partly unfolding are all the reason of GI inactivation in denaturants. PMID- 12552885 TI - [The study of ethanol-water used as reversed-phase high performance liquid chromatographic mobile phase]. AB - The viscosity of ethanol-water at various temperatures was determined. The physicochemical properties and characters of the solvents used as the mobile phase of reversed-phase high performance liquid chromatography(RP-HPLC) were compared between methanol and ethanol. The components of several Chinese medicines have been determined by RP-HPLC with ethanol-water used as mobile phase, and the results were compared with those obtained with methanol-water or acetonitrile-water. It showed that ethanol can be used as RP-HPLC mobile phase by increasing the column temperature moderately. PMID- 12552886 TI - [Simultaneous determination of gibberellins A1 and A3 in foxtail millet seedlings by high performance liquid chromatography]. AB - Gibberellin A1(GA1) and Gibberellin A3(GA3) in millet seedlings were simultaneously determined by HPLC on a Zorbax-ODS-C18 (4.6 mm x 250 mm, 10 microns) column, using a mobile phase consisting of V (0.02 mol/L phosphate buffer solution, pH 4.0):V(methanol) = 2.2:1, a flow rate of 0.8 mL/min and a UV detection at 210 nm. GA1 and GA3 were separated completely in 10 min. Correlation coefficients for linear equations were 0.9986 and 0.9978, recoveries were 102.54% and 99.59%, respectively. Two normal cultivars and their dwarf-mutants were determined by this method. The results showed that both GA1 and GA3 were discovered in foxtail millet seedling, and endogenous mass ratio of GA1 and GA3 in foxtail millet dwarf-mutants were significantly lower than those in normal cultivars, and the concentrations of endogenous GA1 and GA3 were important factors for plant-height. PMID- 12552887 TI - [Instrumental analysis and systematic investigation on heavy oils from coal. II. Analysis of aromatic ring distribution by high performance liquid chromatography]. AB - In this paper, the relationship of chromatographic retention index (I) and aromatic ring number (ARN) of 49 pure reagents in Zorbax-NH2 column/hepatane chromatographic system using high performance liquid chromatography(HPLC) have been studied. The results obtained were that log I values can be divided into several regions according to ARN. They were 0.672-1.089, 1.652-2.588, 2.629 3.241, 3.447-4.160 and 4.482-5.000 for ARN 1 through 5. Those were clear-cut and could be applied to the division of aromatic ring distribution (ARD) of heavy oil products. Hence the ARD method, which was based on the results from pure synthetic compounds, has been established. On the study of quantitation, the peak area normalization method was used to quantify the aromatics with different ARN. The method has the features of high rapidity, high resolution and long life of column. In this paper heavy oil and pitch products from seven plants have been systematically analyzed for their ARD. PMID- 12552888 TI - [Separation and determination of the ingredients of cold medicines by capillary electrophoresis with amperometric detection]. AB - Determination and separation of three ingredients--chlorphenamine maleate, acetaminophen and vitamin C in cold medicines by capillary electrophoresis with amperometric detection were carried out. The effects of detection potential and organic modifier were extensively investigated. Separation efficiency could be improved by adding 1, 2-propanediol into buffer. On optimizing condition, chlorphenamine maleate, acetaminophen and vitamin C were separated satisfactorily and eluted within 12 min. The detection limits were 0.02, 0.08 and 0.01 mg/L respectively. The ingredients in several cold medicines were detected successfully. The recoveries of the samples were 94.2%-102.8%. The method is simple, rapid and highly reliable for routine analysis. PMID- 12552889 TI - [Ion chromatography of L-ascorbic acid, sulfite and thiosulfate using their postcolumn reactions with cerium (IV) and fluorescence detection of cerium (III)]. AB - An ion chromatographic method was used to separate the species of L-ascorbic acid, sulfite and thiosulfate in their mixtures. This method is based on the separation of each anion in their mixtures by using a separation column, and then on the fluorimetric measurement of cerium (III) formed by a postcolumn reaction of cerium (IV) with the species of L-ascorbic acid, sulfite and thiosulfate in the effluent. The optimal conditions for separating and determining the above three species have been established. By using a 3 mmol/L carbonate eluent, the species of L-ascorbic acid, sulfite and thiosulfate could be eluted at the proper retention times of 1.7, 2.6 and 5.0 min, respectively, and these three anions could be separated completely. The effects of the concentrations of cerium (IV) and sulfuric acid in the postcolumn reaction solution on the chromatographic peak height were tested in order to obtain the optimal peak-height. It was found that the peak-height at first increases rapidly with an increase in the concentration of cerium (IV) and sulfuric acid respectively up to a certain concertation, then increases slowly. These critical concentrations of cerium (IV) and sulfuric acid also depend on the amount of the analyte injected. Meanwhile the baseline signals of the sepectra increase with an increase in the concentration of cerium (IV). Some concentrations above the critical concentration of sulfuric acid could be selected as the optimal concentration of sulfuric acid, but the concentration of cerium (IV) should be optimized by establishing a compromise between the higher peak-height and the lower baseline signal. The detection limit of this method was found to be 1 mumol/L for thiosulfate when an amount of 100 microL analyte was injected. PMID- 12552890 TI - [Analysis of organic acids in human dental plaque by means of gas chromatography/mass spectrometry]. AB - The purpose of this study was to investigate the composition of plaque fatty acids in the healthy population. The study was made on 10 volunteers over the age of 18 who were divided into three sub-groups (3-4 individuals). Neither subject exhibited clinical evidence of salivary gland disorder and any medication affecting salivary functions were not used. A sensitive GC/MS method with VG7070E mass spectrometer was developed in our study. The sample separation was carried out on a fused silica capillary column with OV-1. The column size was 23 m x 0.23 mm. The temperature program was as follows: from 40 degrees C to 120 degrees C fast, then from 120 degrees C to 240 degrees C at 6 degrees C/min. The results showed that there were 14 organic acids and isomers present in plaque. They were C12:0, C14:0, C15:0, C16:0, C16:1, C18:0, C18:1, C18:2, C20:0 and phenylacetic acid, phenylpropionic acid. The higher content of fatty acids in the sample were C16:0, C18:0, and C18:1. The aromatic acids were detected only in some samples. The odd-numbers fatty acids and aromatic acids were for the first time detected. The origin of organic acids in plaque is an ongoing area of research. Our data clearly identify the bacterial contribution to the organic acids of plaque matrix, which may have a special relationship with bacteria metabolism. The research will help us to understand fatty acids metabolism of dental plaque and to determine their role in the microbial homeostasis of dental plaque. PMID- 12552891 TI - [Determination of dyes in cosmetic by high performance liquid chromatography]. AB - A reversed-phase high performance liquid chromatographic method was used to detect 5 dyes including p-phenylenediamine in cosmetrics. A Zorbax C8 column was used and mobile phase was V (triethanolamine):V(water):V(acetonitrile) = 0.95:94.05:5 adjusted to pH 7.7 by phosphoric acid. The wavelength 280 nm was selected. Samples were extracted with 95% ethanol by ultrasonic method. The recoveries were 87%-107% and the relative standard deviations were 2.3%-6.4%. PMID- 12552892 TI - [A method of following analysis for glutaraldehyde preparation]. AB - A GC method for the products in cyclopentene oxidation to form glutaraldehyde in a liquid-liquid phase transfer reactive extraction process, which was a new process, is presented. A model GC-103 gas chromatograph equipped with FID was used. The column is a SS tube(phi 3 mm x 4 m) packed with 13% SE-30 coated on 101 white silanized support and a SS tube(phi 3 mm x 2 m) packed with 9.1% PEG-20M coated on the same support. Operating parameters: column temperature programming, 70 degrees C(4 min), then 10 degrees C/min to 140 degrees C(6 min); injector temperature, 185 degrees C; detector temperature, 185 degrees C; N2, 45 mL/min; H2, 65 mL/min; air, 300 mL/min. By using this method, cyclopentene, intermediates, solvents, glutaraldehyde, and byproducts were separated properly. The yield of glutaraldehyde determined by this method is more reliable than that determined by the chemical method. PMID- 12552893 TI - [High performance liquid chromatography for analysis of reverse transcription polymerase chain reaction (RT-PCR) products]. AB - The polymerase chain reaction is a powerful method for amplifying specific DNA sequences in vitro. Reverse transcribing mRNA into cDNA expands the use of PCR to monitor mRNA expression in biological system. A method for the analysis of RT-PCR products by HPLC was developed. The separation was performed on a nonporous ion exchange resin column with gradient elution of sodium chloride in 20 mmol/L Tris HCl buffer (pH 9.0) at a flow rate of 1.0 mL/min and the detection wavelength was 260 nm. lambda-DNA-Hind III digest and a series of RT-PCR products were analyzed for studying the mRNA expression of secreted phospholipase A2 after being injured. PMID- 12552894 TI - [Determination of glycyrrhetic acid in toothpaste by reversed-phase high performance liquid chromatography (RP-HPLC)]. AB - A method for determining glycyrrhetic acid in toothpaste by RP-HPLC is described. The operating conditions were YWG-C18 column (4.0 mm i.d. x 250 mm, 10 microns) at room temperature, V(methanol):V(0.01 mol/L KH2PO4) = 85:15 (pH 3.0) as mobile phase with flow rate of 1.0 mL/min and UV detection at 254 nm. Glycyrrhetic acid in toothpaste samples were extracted with methanol. The average recoveries of glycyrrhetic acid were 99.61%-101.67%. The relative standard deviations were 1.85%-3.16%. The method is simple, rapid and accurate. PMID- 12552895 TI - [Determination of eleven preservatives including bronopol in cosmetic by reversed phase ion-pair chromatography]. AB - Reversed-phase ion-pair chromatography was used to detect 11 preservatives including bronopol, esters of p-hydroxybenzoic acid (PHBA) in cosmetics. Zorbax C8 column was used and mobile phase was V (0.05 mol/L NaH2PO4):V(methyl alcohol):V(acetonitrile) = 50:35:15 with addition of hexadecyltrimethylammonium chloride (2 mmol/L) and the pH adjusted to 3.5 by using phosphoric acid. The wavelength 254 nm was selected for detection. Samples were extracted with methanol by ultrasonic method. The recoveries were 85%-107% and the relative standard deviations were 2.2%-7.5%. PMID- 12552896 TI - [Determination of diosgenin in Rhizoma Paridis by high performance liquid chromatography]. AB - A method for the separation and determination of diosgenin in Rhizoma Paridis by reversed-phase high performance liquid chromatography was developed. The Rhizoma Paridis powder samples were extracted with methanol in a Soxhlet extractor. After extraction the solvent was evaporated and the extract was hydrolysed with 2 mol/L hydrochloric acid for 2 h in a boiling water bath. Then the diosgenin was extracted with petroleum ether (b.p. 60-90 degrees C). The operating conditions were Symmetry C8 column (5 microns, 3.9 mm x 150 mm) at 30 degrees C, mobile phase of V(acetonitrile):V(water) = 75:25 and UV detector at 203 nm. The linearity of the calibration curve was good in the range of 2.1-10.5 micrograms for diosgenin(r = 0.9994). The average recovery and RSD of diosgenin were 99.1% and 1.7% (n = 5) respectively. The method is accurate and reproducible and has been applied to the analysis of Rhizoma Paridis from different sources. PMID- 12552897 TI - [Study on proteins of allele by using the microisoelectric focusing]. AB - Clear protein electrophoresis pattern of single pollen grain from Cucurbita pepo L. was obtained with isoelectric focusing and supersensitive stain technique. The results showed that there were two special distributing protein bands near acidity pole. In species I, the two protein bands were a little near to acidity pole and in species II, that was a little far from acidity pole. In species III, the two protein bands distributed either like species I or like species II and the distributed ratio was 1:1. Moreover, we discuss how to identify relative purity of gene with the sorts of the special proteins. PMID- 12552898 TI - [Analysis of acidic and cationic dyes by thin layer chromatography]. AB - This paper describes the selection of solvent system of thin layer chromatography(TLC) by method of the correlation coefficient for the separation of acidic and cationic dyes. The acidic and cationic dye mixture was extracted from the same coloured wool fiber and acrylic fiber samples. The two solvent systems with smallest correlation coefficients could offer more informations for the identification of the dyes. The method is simple, accurate, effective and can be applied to examine dyes on the textile by selection of TLC solvent system in the forensic science laboratories. PMID- 12552899 TI - [Analysis of physicochemical property and composition of fatty acid of almond oil]. AB - Almond oil was extracted from almond and saponified with saturated KOH-methanol. The mixed fatty acids methylated by methanol-H2SO4(4:1, V/V). Methyl esters were extracted by ether and detected by gas chromatography. The major fatty acids in Taiyuan almond oil were found to be about 68% oleic acid (C18:1), 25% linoleic acid(C18:2), 4.6%-4.8% palmitic acid(C16:0) and a little of palmitoleic acid(C16:1), stearic acid(C18:0). A trace of arachidic acid(C20:0) was also found. PMID- 12552900 TI - [Analysis of 2-acrylamido-2-methyl propane sulfonic acid by thin layer chromatography]. AB - A method for the analysis of 2-acrylamido-2-methyl propane sulfonic acid(AMPS) by TLC is described. The plate was Silica GF254. We selected benzene/methanol as developing agent from several developing systems and fixed the best ratio: V(benzene):V(methanol) = 2:1. The experiment indicated that the Rf value of AMPS was 0.36 while that of AM(acrylamide), the main impurity in crude AMPS sample, was 0.64. The minimum detectable limit of AMPS was 5 micrograms and the developing time was about 25 minutes. Compared with ether/methanol and chloroform/methanol, benzene/methanol system has the advantages of more easily controlling its ratio, better repeatability and much rapid developing rate. This method may be helpful in the control of AMPS in quality. PMID- 12552901 TI - [Cloning and expression of Trichoderma reesei endoglucanase III (EG III) gene in Saccharomyces cerevasiae]. AB - Using Congo red-staining method, one positive clone with CMCase activity was isolated from the Trichoderma ressei cDNA gene bank constructed in Saccharomyces cerevasiae. Sequencing result showed that the 1.5 kb-length DNA fragment inserted in the recombinant plasmid encoded EG III gene from T. reesei. Enzymatic characterization of the EG III produced by recombinant S. cerevasiae was analyzed. The experimental results indicated that the optimum pH and temperature for EG III are 5.0 and 60 degrees C, respectively. The effects of secretory system components SSO 2 and SEB1 of S. cerevisiae on EG III secretion were examined. The results indicated that the amount of EG III secreted by the strain with SSO 2-overexpression was highest among the different recombinant S. cerevisiae strains, showed that SSO 2 is a rate-limiting component of the secretory machinery in the process of EG III secretion. Furthermore, the EG III expression level was increased 5.3 times by deletion. Furthermore, the EG III expression level was increased 5.3 times by deletion of the 98 bp in 5' untranslated region of eg3 mRNA sequence. This result suggested that the regulation region could exist in the 5' untranslated region of EG III mRNA, which is recognized by the gene expression related factors of S. cerevasiae. PMID- 12552902 TI - [Identification of Mortieralla isabellina M6-22 delta 6-fatty acid desaturase by heterologous expression in Saccharomyces cerevisiae]. AB - Using plasmid pTMICL6 containing delta 6-fatty-acid desaturase gene from Mortieralla isabellina M6-22 as a template, 1.38 kb DNA fragment was amplified by PCR. The fragment was subcloned into the yeast-E. coli shuttle vector pYES2.0, then an expression recombinant plamid pYMID6 containing target gene was constructed. The pYMID6 was transformed into Saccharomyces cerevisiae for expression by LiAc method. It was found to exhibit delta 6-fatty acid desaturase activity in the recombinant S. cerevisiae YMID6 in the presence of exogenous fatty acid substrate linoleic acid under introduction of GAL. Expression of the delta 6-fatty acid desaturase gene under appropriate media and temperature conditions led to the production of gamma-linolenic acid reached 8.69% of the total yeast fatty acid by GC-MS detection. It is the first report about expression of M. isabellina D6D gene in S. cerevisiae. PMID- 12552903 TI - [Cloning and sequencing of the gene encoding esterase estA from Ralstonia eutropha CH34]. AB - An esterase-positive clone was isolated by screening a genomic library of Ralstonia eutropha CH34 constructed in Escherichia coli S17-1 with top agar containing alpha-naphtyl acetate and Fast Blue RR. A gne encoding esterase activity, estA was subcloned from this clone. Nucleotide sequencing of estA showed that it was a 825 bp open reading frame, encoding an esterase EstA, composed of 275 amino acids with a predicted molecular mass of 30,785 D. Homology analysis revealed that EstA exhibited significant amino acid similarity with the enzymes involved in the meta-cleavage pathway in the metaboleism of aromatic acid compounds. PMID- 12552904 TI - [Cloning and phylogenetic analysis of hepatitis-associated TT virus subgenome]. AB - Polymerase chain reaction (PCR) was utilized for the DNA amplification from transfusion transmitted virus (TTV) positive serum samples. Five TTV DNA fragments, overlapped about 90% of the genome, were amplified by long template PCR for the generation of TTV subgenome. Recombinant plasmids were obtained by directly inserting PCR products into pT-Adv vector, and DNA sequence analyses showed they were TTV DNA fragments. By using specific restriction enzymes, five TTV DNA fragments were ligated into a TTV DNA subgenome clone and named as TTV021. TTV021 has been deposited in GenBank database with the accession number AF254410. The results of computer analyses showed that TTV021, 3472 nt long, contains two open reading frames (ORF1, 785 aa; ORF2, 146 aa). Identity alignments between TTV021 and other TTV isolates indicated several high conserved regions existed. Phylogenetic analysis of 356 nt from TTV021 suggested that the isolate has close evolutionary relationship with CHN1 (type 1a), but has far relation with other TTV isolates. PMID- 12552905 TI - [Cloning and expression of two garlic virus coat protein genes]. AB - The coat protein(CP) genes of garlic mosaic virus(GMVc) and garlic latent virus(GLVc) isolated from garlic(Allium) plants in Tianjin, China, were amplified from an established cDNA library by PCR method and subsequently expressed in E. coli. using the pET-30a expression system. The determined sequences of GMVc and GLVc CP genes show that the complete GMVc CP gene has 867 nucleotides encoding 289 amino acids. It has 88.5% and 97.2% homology, at the levels of nucleotide and amino acid, respectively, to a reported GMV, indicating that it belongs to Potyvirus. The complete GLVc CP gene has 885 nucleotides coding for 294 amino acids. It has 73.6% and 90.9% homologous percents, in nucleotide and amino acid, respectively, compared to a previously reported GLV, suggesting that it is a member of Carlavirus. The expressed products presented in inclusion body and were analyzed by SDS-PAGE. The molecular weights of GMVc and GLVc CPs appear in 32 kD and 34 kD size, respectively, which are consistent with the deduced sizes of these two CPs. These data will be virtually significant to the further investigation of viruses infecting parlic plant, the control of garlic virus diseases and the production of virus-freed garlic plants. PMID- 12552906 TI - [Studies on transference of hydrogenase genes of Rhizobium arachis]. AB - The hydrogen-uptake genes were transferred into wild Rhizobium arachis Ra strains (Hup-, Nif+, Apr) by triparental mating using pRK2013 as help plasmid. A transconjugant R. arachis Rz34-2(Hup+, Nif+, Apr, Tcr) which expressed high activities of hydrogenase and nitrogenase under free-living and symbiotic state was screened. Peanut inoculation test with recipient R. arachis Ra34, transcojugant Rz34-2 and control strain R. arachis L8-3 (Hup+, Nif+) was carried out respectively. The results showed that, compare to treatment without inoculation, inoculation with R. arachis Ra34 and R. arachis L8-3, the dry weight of leaf inoculated with transconjuant Rz34-2 increased 6.2%, 7.6% and 6.3% respectively; the N-content of seed increased 8.8%, 10.0% and 6.0%; the output increased 18.8%, 10.5% and 10.7%. This suggested that legume plants inoculated with Rhizobium strains (Hup+) were more efficient to accumulate N and to increase its output. PMID- 12552907 TI - [Expression of otsA gene in tobacco and improvement stress tolerance]. AB - The Escherichia coli trehalose-6-phosphate synthase gene(otsA) was engineered under the control of cauliflower mosaic virus regulatory sequences (CaMV35S) for expression in plants. OtsA gene was incorporated into the chromosome DNA by Agrobacterium-mediated transfer and expressed in Nicotiana tabacum. The otsA gene transgenic plant exhibited multiple phenotypic alteration: improved stunted growth and drought tolerance. The detached leaves from the transgenic plant, their water loss slower than that of the controls. Trehalose accumulated in transgenic plant was determined by HPLC suggest that synthesis of the sugar improving plant to stress tolerance. PMID- 12552908 TI - [Incompatibility among indigenous plasmids of Sinorhizobium fredii strains and its application for plasmid curing]. AB - The indigenous plasmids of S. frediii HN01 and WWG18 were labeled with Tn5 MobsacB. Those plasmids were cured on TY medium containing 10% sucrose to identify its function. The pSymHN01b of HN01 was transferred into WWG18SR and C361SR. The results of plasmid detection, curing and pot plant experiment revealed that the incompatibility was existed between pSym-HN01b and symbiotic plasmid of WWG18SR or the third plasmid of C361SR. But it was compatible with the nonsymbiotic plasmid of WWG18SR and symbiotic plasmid of C361SR. Using plasmid incompatibility, The symbiotic plasmid of WWG18SR and one nonsymbiotic plasmid of C361SR were eliminated successfully. PMID- 12552909 TI - [Effect of gene disruption of aveD on avermectins production in Streptomyces avermitilis]. AB - Recombinant plasmid pCZ2(pKC1139::475 bp aveD) was used for aveD gene disruption in Streptomyces avermitilis 76-9. The plasmid was inserted into the chromosome by homogenous recombination between partial aveD gene in the plasmid and aveD in the chromosome. Disruptants were confirmed by Southern blotting. Shaking flask experiments and HPLC analysis showed that the disruptant produced only four components, which were C5-oxo-avermectin B1a, B1b, B2a, B2b as identified by UV, IR, NMR, and MS. This revealed that both aveD and aveF were not expressed in the disruptant. This is consistent with that aveD and aveF are in a transcription unit. This paper also provided a new genetic method to obtain C5-oxo-avermectin B producing strain. PMID- 12552910 TI - [Production and detection of monoclonal anti-idiotype antibodies against Vibrio anguillarum]. AB - Vibrio anguillarum is the pathogenic bacteria of Vibriosis, which is an infectious disease found in various fish species. Seven monoclonal anti-idiotype antibodies(mAb2) were raised against mAb1 4A6. Identification of subgroup showed that 1H5, 1D1, 2B12 and 2F12 belonged to IgG2b, 2H12 and 1H12 to IgG2a and 1E10 to IgG3. The titers of these mAb2 ascites were 1 x 10(-4)-1 x 10(-6). The capacity of the mAb2 to inhibit the binding between mAb1 and antigen was investigated with the competitive inhibition ELISA. The results showed that 1D1, 1E10 1H5 and 2H12 mAb2 were able to inhibit this binding. Another experiment demonstrated that four mAb2(1D1, 1E10, 1H5 and 2H12) might induce Balb/c mice to produce Ab3 and these Ab3 competed the same antigen epitopes with Ab1. These results indicate that mAb2(1D1, 1E10, 1H5 and 2H12) are likely to represent internal image of antigen and belong to Ab2 beta. They might be employed to induce antibodies against pathogenic epitopes of V. anguillarum in vivo so as to gave the safe and effective vaccine. PMID- 12552911 TI - [Construction of the full-length cDNA clone of Chinese virulent strain--F114]. AB - Seven subclones covered the complete genome of classical swine fever virus Chinese virulent strain F114 were obtained by reverse transcription PCR. The complete nucleotide sequence of the genome of strain F114 was determined by sequencing. The cDNA fragments were then assembled and inserted downstream of a T7 promoter in pBluescript II sk+ plasmid vector to obtain the full-length cD-NA clone sk-12297. Homology comparison of the nucleotide and amino acid sequence of strain F114 with the known sequences of HCLV, Brescia and Alfort showed 95.70%, 96.80%, 86.03% identity in nucleotide and 97.41%, 98.54%, 93.33% identity in amino acid respectively. PMID- 12552912 TI - [Production and some properties of a beta-glucanase from Trichoderma sp. GXC]. AB - The factors affecting Trichoderma sp. GXC for beta-glucanase (1,3-1,4-beta-D glucan 4-glucanohydrolase, EC 3.2.1.73) production have been investigated, the optimal conditions were that carbon and nitrogen sources were bran and ammonium sulfate, respectively, initial pH 4.0-5.0, shaken at 30 degrees C for 44 h. beta glucanase was purified to electrophoretic homogeneity by using ammonium sulfate precipitation, Sephadex G-100 and DEAE-Sephadex A-50 chromatography. It's molecular weight was 35 kD by SDS-PAGE, and the enzyme functioned optimally at pH6.0 and 50 degrees C, respectively. In addition, the enzyme was stable at pH 4.0-5.0 and below 40 degrees C. The beta-glucanase activity was significantly inhibited by Fe3+ and stimulated by Co2+, respectively. PMID- 12552913 TI - [Purification and characterization of 2-carbonyl reductase from marine bacteria Bacillus sp]. AB - ANADPH-dependent 2-Oxoaldehyde reductase was isolated and purified from a marine bacteria Bacillus sp. The purification procedure involved ammonium sulfate fractionation and Q Sepharose FF, Hydroxyapatite, Sephadex G-100 column chromatographies. The specific activity of the purified enzyme was increased by 141.1 folds over crude extract and the recovery yield was 11.4%. 2-Oxoaldehyde compounds were found to be speciall good substrates. The optimum pH of the enzyme activity was 6.2-6.6. The Km coefficient for 3-deoxyglucosone was 2.5 mmol/L. The molecular weight of the enzyme was estimated to be 33 kD The enzyme activity is stable below 30 degrees C and pH 5.0-8.0. EDTA, beta-mercaptoethanol and dithiothreitol enhanced the enzyme activity. On the other hand, the enzyme activity was partially lost by idoacetic acid or N-ethylmaleimide. PMID- 12552914 TI - [Purification and properties of endoglucanases from Aspergillus aculeatus SM L22]. AB - The five endoglucanases(CMCase) components from Aspergillus aculeatus SM-L22 were separated and purified by exclusion chromatography and ion-exchange chromatography. Five components(EG II-1, EG II-2, EG III-1, EG III-2 and EG IV) had molecular weights of 38.7, 34.4, 31.4, 36.9 and 23.7 kD by SDS-PAGE, respectively, and IEF showed their pI were pH < 3.5, < 3.5, 4.9, 4.4 and 5.0, respectively. All of them have maximum reactive activities at pH 3.5-4.0; and the optimum temperatures were 55 degrees C, 60 degrees C, 60 degrees C-70 degrees C, 60 degrees C-70 degrees C and 60 degrees C, respectively. EG II-1 and EG II-2 can only act such morphological substrates as CMC or phosphated cellulose, but EG III 1, EG III-2 and EG IV can active xylan also. The activities of all components were stimulated by Fe2+ except EG IV, EG III-2 was activited mostly by Fe2+. The kinetics' showed that there were no relativies between the affectivity of cellulases and its Km. PMID- 12552915 TI - [Immobilization of penicilin G acylase on polyacrylonitrile fiber]. AB - The immobilization of Penicillin G Acylase from Bacillus megaterium by glutaraldehyde crosslinking on the partially acid-hydrolyed polyacrylonitrile fiber was studied. When the amount of--NH2 on fiber were 690 mumol/g and the moisture in the fiber was 64%, and the content of enzyme protein immobilized on fiber was more than 100 mg/g. The activity of 2300 IU/g was obtained with 30% of overall yield and 56% of binding efficiency. The immobilization yield was markedly influenced by ratio of the amount of free enzyme used to the weight of the fiber. The half-life of storage stability of immobilized PGA at room temperature was 130 days. The immobilized PGA kept 80% of the initial activity after 20 cycles of operation in 10% of PGK(W/V) in 0.05 mol/L phosphate buffer, pH 8.0, at 37 degrees C and an enzyme load of 150 IU/g(PGK) and 10 g(PGK) for per cycle of operation. The hydrolysis conversion of PGK in the range of 2.5%-12.5% (W/V) were over 98% for the immobilized PGA. The operation stability of immobilized PGA treated with DTT was better than that of immobilized PGA untreated. PMID- 12552916 TI - [Studies on regulation of glutamine synthetase activity from Streptomyces lincolnensis]. AB - Glutamine synthetase in crude extracts from Streptomyces lincolnensis growing under different nitrogen sources were studied. The results showed that NH4+ in high concentration repressed the biosynthesis of the enzyme. To determine whether Streptomyces lincolnensis has undergone covalent modification, a comparison of the glutamine synthetase isolated from cells grown on different nitrogen sources was made. No significant difference was observed in specific activity, pH optima, divalent cation response, and ultraviolet absorption spectra. Glutamine synthetase activity was not influenced by ammonia shock or snake venom phosphodiesterase treatment. Under these conditions, the activity of glutamine synthetase from K. aerogenes was markedly changed. There was therefore no evidence for enzymatic adenylylation of glutamine synthetase from Streptomyces lincolnensis. Glutamine synthetase was subject to feedback inhibition by end products of glutamine metabolism. Cumulative feedback inhibition of the Mn(2+) dependent glutamine synthetase activity was demonstrated. These results suggest that glutamine synthetase from Streptomyces lincolnensis is an allosteric enzyme. PMID- 12552917 TI - [Effect of microgravity simulation with clinostat on different strains in Anabaena oryza]. AB - The Biological responses of Anabana oryza (different strains) to microgravity simulation with clinostat were analyzed. The simulation of growth rate was observed under clinoration all the strains, especially in flight strains (AoR16 and AoSR16-17). Anabana oryza HB23 showed higher activity than flight strains in photosynthesis and respiration. The nitrogen which flight strains with high nitrogrnase activity fixed under culturing in ground was used to accumulate phycobiliprotein in cell and excrete ammonium out of cell after meeting the normal metabolism, however, it was mainly used to simulate growth rate under culturing with clinostat. menwhile, it was different from space flight that microgravity simulation with clinostat was not able to induced the phenotype segregation in Anabana oryzae. PMID- 12552918 TI - [Isolation and identification of the broad spectrum antimicrobial peptide AP311 produced by Lactobacillus acidophilus]. AB - The heat stable antimicrobial peptide (AP311) produced by Lactobacillus acidophilus was isolated and identified. The AP311 has broad spectrum of inhibition including many Gram-positive and Gram-negative bacteria. The inhibition activity of AP311 was lost upon treatment with trypsin, subtilisin, proteanase K, chymotrypsin and pepsin. Inhibition activity of AP311 was decreased with pH increasing. The AP311 is very stable at acidic condition (pH2-4) even if heating at 100 degrees C for 30 min. But it was inactivated at basic pH (pH12) and the activity was restored completely upon reversion to acidic. The AP311 was not precipitated and inactivated by various organic solvents, except n-butanol. When the precipitate was redissolved in 0.02 mol/L HC1, its activity was restored. Based on its proteinaceous nature, broad spectrum of inhibition, we propose that AP311 should be considered the broad spectrum antimicrobial peptide. PMID- 12552919 TI - [Studies on antimicrobial activity of extracts from thyme]. AB - The extracts from thyme by water and ethanol, thyme essential oil, thymol and carvacrol were used as antimicrobial agents in this paper. The results show that all antimicrobial agents used have strong inhibition activity against Staphalococcus aureus, Bacillus subtilis, Escherichia coli. PMID- 12552920 TI - [Effect of drying and temperature on the sporulation of Pandora delphacis mycelia gelatinized]. AB - The entomophthoraceous fungus, Pandora delphacis, is a microbial agent highly potential for control of sucking-type insects. In this study, effort was made of gelatinizing the mycelia of the isolate F95129 from submerged culture using polyacrylamide-starch gel pwder and sodium alginate. The resulting film-like gel of the mycelia sporulated very well, indicating that the materials used for gelatinization of the mycelia was biologically compatible with P. delphacis. Then, the gelatinized mycelia were slowly dried in silica chamber at 4, 14, and 24 degrees C, respectively. After 35-day drying and storage in the chamber, however, a 22.7%-92.8% decrease of sporulation capacity occurred in the film-like gels, which then had different water content. The nutrient-supplemented polyacrylamide-starch gel stored at 14 degrees C with water content of 6.1%-7.4% was best for largely decreasing the loss of sporulation capacity among the combinations of gel, nutrition, water content and temperature. The results indicate that it could be possible to choose a desirable gel for longer biological activity of gelatinized P. delphacis mycelia. PMID- 12552921 TI - [Vibrio cholerae VPI phi/CTX phi/TCP: interactions of phage-phage-bacterium]. PMID- 12552922 TI - [Taxonomy of the nocardioform actinomycetes]. PMID- 12552924 TI - Core public health functions: 15-year update. PMID- 12552923 TI - [Recent researches on fungal hydrophobins]. PMID- 12552925 TI - Reinvigorating public health core functions: restructuring Los Angeles county's public health system. AB - In 1997, the Los Angeles County Department of Health Services launched an effort to revitalize its Public Health Services division. County investments resulted in the creation of 241 new positions, facilitating the establishment of new offices addressing neglected chronic disease prevention, staff development, training, and communication. Service effectiveness and community responsiveness were enhanced by decentralizing leadership through eight Area Health Offices and strengthening partnerships with private health care providers, community-based organizations, and managed care organizations. Infrastructure enhancements included the development of program performance measures, countywide health indicators, a central health assessment and epidemiology office, and a new countywide biennial survey. PMID- 12552926 TI - The reinvigoration of public health nursing: methods and innovations. AB - Los Angeles County (LAC) restructured and reinvigorated public health in response to nationwide concern over the adequacy of all public health infrastructures and functions. LAC's reorganization into geographically defined service planning areas (SPAs) has facilitated the integration of core public health functions into local practice. Public health nurses practicing as generalists within their SPA identified three initial objectives to address in population-based care: (1) expanding practice beyond disease control to a more holistic approach, (2) providing consultation using the Ask-the-Nurse innovation, and (3) developing a community assessment database for interdisciplinary SPA health planning. Additional innovative objectives are planned for the future. PMID- 12552927 TI - Local health department performance: compliance with state statutes and rules. AB - Compliance with statutes and rules is a measure of local health department (LHD) performance. This study analyzed compliance rates in one state. High overall compliance rates were found, although gaps existed between measured and expected compliance. Factors associated with lower compliance included lower LHD certification level (characteristic of smaller agencies) and human services organizational structure in lower-level LHDs. Improvements in compliance rates will require increased attention to development and documentation of policies, procedures, and local ordinances. Compliance is necessary for LHD effectiveness but may not be a sufficient measure of overall LHD performance. PMID- 12552929 TI - Systems analysis of a clinical error. AB - Systems analysis is an approach used in situations requiring high reliability. In the past, the investigation and prevention of clinical error focused on the actions of individuals. A systems analysis approach to medical errors can identify and correct multiple contributing factors resulting in safer patient care systems. Recent analysis of a clinical error that occurred in public health practice identified multiple factors contributing to its occurrence. The use of the systems analysis approach resulted in high staff satisfaction and creative suggestions for operating a safer clinic and providing better patient care. PMID- 12552928 TI - Assuring community emergency care capacity with collaborative Internet tools: the Milwaukee experience. AB - Hospital overcrowding and diversion of ambulances from emergency departments are being recognized as increasing problems in the health care system. This article, a descriptive narrative, examines the various factors contributing to the problem and describes how collaborative approaches to public health issues can be applied. It describes Milwaukee's experience with a collaborative approach. The use of a technological tool to assist with tracking and reporting on ambulance diversion and emergency department overload is explained, and data are provided to show the impact of various methods to blunt the impact of the flu season on diversion frequency. The article encourages use of similar collaborative approaches and Internet-based technology to address other public health problems. PMID- 12552930 TI - Developing performance measures for immunization registries. AB - Quantitative performance indicators were developed to monitor immunization registry progress of the 16 All Kids Count (AKC) grantees. Results were reviewed to determine if (1) the indicators measured immunization registry progress, (2) the results could be compared across AKC projects, and (3) other immunization registries could use the indicators. The study found that the AKC performance indicators provide a useful template for registries to measure their progress toward developing fully mature registries. Public health leaders should join with private health sector representatives to build on the immunization registry experience and develop and test performance indicators for integrated information systems in order to develop common goals and monitor changes in a comprehensive way. PMID- 12552931 TI - Implementing program evaluation and accountability for population health: progress of a national diabetes control effort. AB - Diabetes affects some 16 million Americans at a cost estimated at $100 billion. The Centers for Disease Control and Prevention funds a diabetes control program (DCP) in every state as part of the National Diabetes Control Program (NDCP). In 1999, a new policy added emphasis on evaluation and made NDCP and its DCPs accountable for achieving impacts related to the health of populations with diabetes. The article reports on the experiences of the NDCP in implementing a performance-based program evaluation paradigm. It also discusses potential future directions for national diabetes-control efforts. PMID- 12552932 TI - Establishing a statewide mammography database in Arkansas: overcoming the barriers. AB - To determine patterns of mammography utilization in Arkansas, the Arkansas Mammography Data Collection Project (MDCP) was established. The project's objective was to compile into one database statewide information about mammograms performed. All mammography centers were invited to participate in the project. Many barriers were encountered that were center related, data related, or personnel related; different interventions were implemented for each barrier. At the conclusion of the project, 92 out of 112 centers (82%) participated in the project, creating a database of 157,976 mammography data sets. Identifying and overcoming many of the barriers were crucial steps in the project's success. PMID- 12552933 TI - Media campaign influences parents' opinions about their children and tobacco. AB - The study assessed the effectiveness of a media campaign in convincing parents that children are targeted by the tobacco industry and that children should be banned from tobacco use. An 800-parent sample survey was conducted prior to a statewide anti-tobacco campaign, and another survey sample of 790 parents was conducted afterward. THE RESULTS: Though parents who smoked in high school and those who did not agreed that children are targeted and should be banned from tobacco use, parents who did not smoke in high school believed it more strongly. Parents who had a history of smoking changed more on the issue of banning, perhaps because they had more room to change. The study concluded that media campaigns can change parents' attitudes. PMID- 12552934 TI - The public health workforce in the year 2000. AB - It has been 20 years since the last estimate of the public health workforce's size and composition. This study provides a best estimate through secondary analysis of existing workforce reports, summaries, and information gathered from chief health officials in 50 states and 6 territories, public health organizations, and federal agencies. Results indicate that the workforce consists of some 448,254 persons in salaried positions as follows: 44.6 percent professionals, 4 percent officials/administrators, 14 percent technicians, and 13 percent clerical/support. Workforce size may have decreased by as much as 10 percent over the last 20 years, despite a 25 percent increase in population and rising health hazards. PMID- 12552935 TI - Efficacy of combined radiation, paclitaxel and carboplatin for locally advanced non-small cell lung carcinoma. AB - Locally advanced non-small cell lung carcinoma (NSCLC) has a poor prognosis when treated with conventional chemotherapy and radiation. New chemotherapy agents like paclitaxel may increase the sensitivity of tumors cells to radiation and potentially improve the outcome. The optimal combination of taxane-based chemotherapy agents and radiation is still unclear. We investigated the feasibility of induction chemotherapy followed by concurrent near systemic dose of chemotherapy with radiation. A prospective survey of 29 previously untreated patients with unresectable stage III (15 IIIA, 14 IIIB) NSCLC treated with paclitaxel and carboplatin in combination with radiation was reviewed. The patients received 2 cycles of paclitaxel 225 mg/m2 intravenously (i.v.) over 3 hours, days 1, 22; carboplatin at area under the curve (AUC) 6 based on Calvert formula days 1, 22 following completion of the paclitaxel infusion. Following induction chemotherapy, radiation therapy started on day 43 until completion to a tumor dose of at least 5960 cGy. Cycles 3 and 4 of chemotherapy were begun on days 43 and 63, respectively, and consisted of paclitaxel 175 mg/m2 i.v. over 3 hours, and carboplatin at AUC 6 following paclitaxel infusion. The response rate, acute toxicity, long-term complications, pattern of failure and survival were evaluated and compared to previous studies in the literature. Two patients were lost to follow-up. The response rate to induction carboplatin/paclitaxel was 52%. An overall response rate (complete and partial responders) of 85% was obtained following chemotherapy and radiation. Grade 3-4 acute side-effects were recorded in 9 patients (31%) and consisted of esophagitis (8 patients) and anemia (1 patient). One patient died from cachexia 3 months following treatment (3.7%). The median survival and 3-year survival were 15 months and 30%, respectively, for the remaining 27 patients at a median follow-up of 11 months. There was no difference in survival between stages IIIA and IIIB at 2 years (IIIA: 22%, IIIB: 31%). Local or regional recurrences and distant metastases developed in 9 patients (33%) and 13 patients (46%), respectively. The combination of paclitaxel, carboplatin and radiation for locally advanced non-small cell carcinoma is feasible with acceptable toxicity. The response rate compares favorably with previously reported studies. The decrease of tumor volume following induction chemotherapy allows sparing of the lungs from the toxicity of radiation. However, grades 3-4 esophagitis remain significant. The addition of amifostine may be beneficial in this setting. PMID- 12552936 TI - Age- and site-dependent cell cycle composition of the normal human colonic mucosa. AB - BACKGROUND AND AIM: As a reference to studies of DNA-ploidy and S- and G2/M-phase fractions in patients with inflammatory bowel diseases, we describe the mucosa of normal individuals with respect to age and localization in the colon. MATERIALS AND METHODS: One hundred and sixty-five biopsies from the right, transverse and left colon from 44 subjects (20 men, 24 females, median age 55 years (range 21 80)) who were referred for colonoscopy due to rectal bleeding, diarrhoea or suspicion of neoplasia, but with normal macroscopic and microscopic findings, were analysed by DNA-flow cytometry for ploidy and cell cycle composition. The biopsies were immediately fixed in buffered formalin and then analysed by a method for high quality preparations of cell nuclei without any centrifugation steps, resulting in minimal cell damage and low frequencies of aggregates, making the background levels low in the DNA-histograms. RESULTS: The median S-phase fraction of the biopsies, all diploid, was 2.35% (0.1-8.3). The S-phase fraction increased linearly with age (p = 0.001) and decreased from the right colon (median 2.75% (0.5-8.3)) over the transverse colon (median 2.3% (0.1-6.2)) to the left colon (median 1.9% (0.8-6.5), p < 0.02). The fraction of G2-cells (median 1.1%, range 0.2-5.1) increased significantly with increased S-phase fraction (p < 0.0001). CONCLUSION: DNA-FCM analyses of normal colonic tissue demonstrate an age and site-dependent variation with regard to cell proliferation. This variation has to be taken into consideration when biopsy specimens from chronic colitis mucosa are evaluated. PMID- 12552937 TI - Molecular analyses, morphology and immunohistochemistry together differentiate pleural synovial sarcomas from mesotheliomas: clinical implications. AB - BACKGROUND: Only recently pleural synovial sarcomas (SS) have been definitively identified because of the presence of a characteristic X;18 translocation. SS are sarcomatoid or biphasic malignancies morphologically almost indistinguishable from sarcomatoid or biphasic malignant mesotheliomas (MM). PATIENT AND METHODS: We demonstrated a primary pleural biphasic SS in a patient referred to us has having a biphasic MM. RESULTS: Histology showed a spindle cell tumor with focal epithelioid differentiation, microcalcifications, and a hemangiopericytomatous vascular pattern. Ultrastructurally, the epithelioid tumor cells had few blunt microvilli and occasional intercellular junctions. Immunohistochemically the tumor cells were positive for BCL-2, CD99, CD56, and focally for BerEp4, Pancytokeratin and cytokeratin 5/6. These findings suggested the possibility of SS rather than MM. Detection of the t(X:18) translocation using RT-PCR, Southern blot, and DNA sequencing definitively confirmed the diagnosis of SS. CONCLUSION: The differential between pleural SS and MM requires a high degree of suspicion and molecular analyses because morphology (histology, immunohistochemistry and electron microscopy) is insufficient to definitively distinguish between these two malignancies. This differential is critical because patients with pleural SS can be susceptible to chemotherapy, and accordingly are treated, while patients with sarcomatoid MM are resistant to chemotherapy and accordingly are not treated. PMID- 12552938 TI - Relationship between pharmacokinetics of 5-FU in plasma and in saliva, and toxicity of 5-fluorouracil/folinic acid. AB - BACKGROUND: Dose adaptation based on pharmacokinetic parameters has been shown to decrease toxicity of some 5-fluorouracil(5-FU)-based continuous infusion regimens. PATIENTS AND METHODS: In the present study the relationship between 5 FU pharmacokinetics in plasma and in saliva, and toxicity was investigated in 40 patients receiving the combination of 5-FU 425 mg/m2 and folinic acid 20 mg/m2 daily during 5 consecutive days according to the Mayo-regimen. RESULTS: The overall non-hematological and hematological toxicity, as well as mucositis only, were not statistically significant related to the area-under-the-curve in plasma (AUCp) or in saliva (AUCs), nor to the maximum concentration measured in plasma (Cmaxp) or in saliva (Cmaxs). Although statistically significant, the correlation between the AUCp and AUCs was relatively low, implying that salivary pharmacokinetics are not accurately predictive of plasma pharmacokinetics. CONCLUSION: The conclusion of this study is that the application of pharmacokinetic parameters is not appropriate for identification of patients at risk for developing toxicity from treatment with 5-FU according to the Mayo regimen. PMID- 12552940 TI - High-dose chemotherapy in small-cell lung cancer. AB - Small cell lung cancer (SCLC) is highly sensitive both to radiotherapy and chemotherapy. Given its high chemo sensitivity, even two decades ago, SCLC was one of the first malignancies deemed suitable for maximising the dose and dose intensity with the support of autologous bone marrow (ABMT). On the whole, results were disappointing and the procedure was practically abandoned. Nowadays some interest is again emerging due to improvements in supportive care, such as the availability of hematopoietic growth factors and the peripheral blood progenitor cells (PBPC). Data of 505 patients included in 26 studies were reviewed. About two thirds of these patients had LD (limited disease). Late intensification protocols were used in 311 patients who, however, represented only the 30% of the population initially given conventional chemotherapy. Of the patients not achieving complete remission (CR) after induction, high-dose induced a CR in 39% of the cases. The use of early intensification was reported in 8 studies including 194 patients. The CR rate was 51.5%. Overall, the probability of achieving the CR was 2-3 times higher in LD than in ED (extensive disease). Relapses occurred at the site of the primary in more than half of the cases, showing that the course of the disease was not modified by the use of high-dose chemotherapy. Toxic deaths occurred in 7% of the treated patients, without difference in the two treatment methods. Though the schedules were too variable to draw firm conclusions, the ICE (ifosfamide, carboplatin, etoposide) and the CBP (cyclophosphamide, cisplatin, carmustine) regimens apparently provided better results, with a 2-year survival rate of 30-50% in the LD subset. An european multicenter randomized trial is ongoing. At the present time high-dose chemotherapy is still to be considered experimental treatment, since major problems such as the selection of the patients, doses and timing of chemotherapy and radiotherapy remain unsolved. PMID- 12552939 TI - Update of the role of surgery in the multimodal treatment of MALT gastric lymphomas. AB - BACKGROUND: Primary gastric non-Hodgkin's lymphoma is a well-defined clinicopathological entity with a distinct histological spectrum and indolent course. Its optimal management still remains controversial. In this study we present our experience with surgery being part of the multimodal treatment and we compare our results with those referred to in the world literature. MATERIALS AND METHODS: Sixty-five patients (37 males, 28 females) with gastric MALT lymphoma were retrospectively analyzed. Forty-seven patients underwent curative resection, eleven received chemotherapy as primary treatment, two patients received radiotherapy before any other treatment and five patients received combined chemo radiation therapy. RESULTS: For the early stages of the disease (I-III), radical resection offered a two-year survival rate of 100% and 93%, respectively, and a five-year survival of 85% and 67%, respectively. For those stage I patients, who were managed surgically followed by adjuvant therapy, the two- and five-year survival was 90% and 78%, respectively. For stage II patients for whom chemo radiation therapy followed surgery, the two- and five-year survival was 88% and 65%, respectively. CONCLUSION: In view of the results of our study, surgery is an adequate treatment modality for the early stages of the disease. The addition of radiotherapy or chemotherapy does not improve the overall survival. Conversely, for the advanced stages (II2-IV), primary chemotherapy is the best treatment option with surgery being reserved for cytoreduction or chemo-radiotherapy induced complications. PMID- 12552941 TI - A phase I study of continuous administration of 5-fluorouracil/cisplatin in advanced uterine cervical cancer. AB - BACKGROUND: The aim of this study was to assess the toxicity of a neoadjuvant chemotherapy (NAC) regimen consisting of cisplatin (CDDP) and 5-fluorouracil (5 FU) through 24-hour intravenous continuous infusion on days 1-4 in a Phase I study. PATIENTS AND METHODS: Thirteen patients were recruited for this study. All patients were treated with a regimen consisting of CDDP and 5-FU through a 24 hour intravenous continuous infusion on days 1-4, followed by radical hysterectomy and/or radiation. Each initial dose of CDDP and 5-FU was 20 mg/m2/day and 750 mg/m2/day, respectively, for 4 days. RESULTS AND CONCLUSION: In the third step, seven patients were treated at 25 mg/m2 day CDDP and 1000 mg/m2/day 5-FU, respectively, for 4 days. One of seven patients showed Grade 4 thrombopenia. However, in this dose step, all the patients showed an objective response. Although maximum tolerated doses (MTDs) were not reached, we decided to stop the escalation and to recommend this level for the Phase II study. PMID- 12552942 TI - Sarcomatoid adenocarcinoma of the lung: clinicopathological, immunohistochemical and molecular analyses. AB - BACKGROUND: Biphasic lung cancers with admixtures of adenocarcinoma and sarcomatoid components but lacking true mesenchymal differentiation (sarcomatoid adenocarcinomas) are rare, and extensive studies of their clinicopathological characteristics and histogenesis have not been performed. MATERIALS AND METHODS: Six surgically resected sarcomatoid adenocarcinomas were compared clinicopathologically with 317 standard adenocarcinomas, and studied immunohistochemically and by loss of heterozygosity (LOH) analysis focusing on each component. RESULTS: In comparison with standard adenocarcinomas, the sarcomatoid adenocarcinomas occurred in older patients, were more likely to be associated with smoking and were more acinar than papillary, with a worse five year prognosis. Immunohistochemically, sarcomatoid components were positive for epithelial markers in three cases. In one case the carcinoma showed retention of heterozygosity and the sarcomatoid components allelic loss, while all other cases showed retention or LOH in both components. CONCLUSION: The sarcomatoid adenocarcinoma exhibits different clinicopathological characteristics from standard adenocarcinomas. The present immunohistochemical and LOH analyses provided support for the idea that both components are derived from one precursor cell, and that progression may be by way of adenocarcinoma to sarcomatoid differentiation. PMID- 12552943 TI - A case of pulmonary adenocarcinoma associated with hypertrophic osteoarthropathy due to vascular endothelial growth factor. AB - A 62-year-old male patient was admitted, because a chest X-ray examination revealed an abnormal shadow (4 cm in diameter) in the left upper lobe. A transbronchial lung biopsy showed the features of adenocarcinoma. Chest and abdominal computed tomography (CT) scan showed the features of lymphangitis carcinomatosa, and bilateral metastatic adrenal tumors. After effective chemotherapy, he complained of arthralgia and a bone scintigram showed abnormal uptakes in his bilateral shoulder, knee and wrist. He was diagnosed with pulmonary hypertrophic osteoarthropathy (PHOA). The serum level of vascular endothelial growth factor (VEGF) was elevated to 424 pg/ml, while serum growth hormone (GH) and GH-releasing hormone were normal. Immunohistochemical analysis showed positive features of VEGF in the pulmonary tumor cells. He died of cachexia on May 2nd, 2001, more than 2 years after the diagnosis of pulmonary adenocarcinoma at stage IV. PMID- 12552944 TI - Prognostic value of glucose transporter 1 expression in patients with hypopharyngeal carcinoma. AB - BACKGROUND: The increase of glucose transport is a distinguishing marker of tumor cells. We hypothesized that a hypopharyngeal carcinoma, characterized by high frequency of lymph node spread or distant metastasis, was associated with glucose transporter 1 (Glut 1) overexpression. MATERIALS AND METHODS: We investigated the incidence of Glut1 expression by immunostaining in 99 patients with hypopharyngeal carcinoma, and revealed the correlation with the clinical characteristics. RESULTS: The analysis showed 46 patients (46%) of Glut1 overexpression (> 70% tumor cell stained). Glut1 expression did not correlate with gender, subsite, histological grade, tumor size, lymph node status or stage grouping. The 5-year relapse-free survival (RFS) rate of patients with Glut1 overexpression (> 70%) (35%) was significantly lower than that of patients with low Glut1 expression (< or = 70%) (57%) (p = 0.047). Multivariate analysis showed that advanced tumor stage (stages III and IV) and Glut1 overexpression were predictors of reduced RFS (p = 0.031 and 0.021, respectively). CONCLUSION: Glut1 expression is an indicator for predicting the prognosis of hypopharyngeal carcinoma. PMID- 12552945 TI - SV40 in human tumors: new documents shed light on the apparent controversy. AB - BACKGROUND: Presently there are over 61 reports from 49 different laboratories that have detected SV40 in human mesothelioma, lymphoma, brain and bone tumors, versus three reports (two from Dr. Shah's laboratory who performed his study under contract from Dr. Strickler at the Viral Epidemiology Branch (VEB) National Cancer Institute (USA) that have failed to detect SV40 in some of these same tumor types. To address whether the negative reports were caused by lack of sensitivity of the technique used in Shah's laboratory, or whether the positive reports were caused by contamination within the greater number of laboratories reporting SV40 detection, two multi-center studies were conducted. The first study, Testa et al., 1998, confirmed the presence of SV40 in mesothelioma. The second study, Strickler et al., produced irregular results indicating that: (a) though never reporting SV40 detection to date, Dr. Shah's laboratory reported the most sensitive technique of all participating laboratories; (b) all participating laboratories essentially agreed the DNA extracts provided under contract to the VEB were negative; (c) all participating laboratories agreed one-half of the negative controls prepared by the VEB contract laboratory were positive due to contamination by the contract laboratory. In addition, (d) the authors concluded the laboratories previously detecting SV40 in human tissue specimens were not reporting contamination. Scientists in the field have since debated how these seemingly contradictory results were produced. MATERIALS: During the course of litigation representing patients with SV40-positive tumors, the author obtained correspondence among members of the VEB multi-center study and sworn testimony by Dr. Shah that address some of the incongruities of the study. RESULTS: Dr. Shah's laboratory technique used in 1996 was apparently not sufficiently sensitive to detect SV40 in human tumors. When this became apparent, during unilateral pre trial testing of positive controls by Dr. Shah, the study coordinator of the VEB, Dr. Strickler, apparently compromised the blinded nature of the study and allowed Dr. Shah to modify and improve his technique. When one of the participating laboratories questioned irregularities in the data from Dr. Shah's laboratory and directly questioned Dr. Strickler, the study organizer, about the potential irregularity, Dr. Strickler and Dr. Shah offered letters stating that such irregularities had not occurred and re-confirmed that they had not deviated from the standard protocol. CONCLUSION: The facts indicating that Dr. Shah's laboratory technique was not sufficiently sensitive to detect SV40 were not made available to the other laboratories participating in the study and were not published. Instead, according to Dr. Shah's testimony, Dr. Strickler, the VEB multi-center study coordinator, compromised the masked positive controls and knowingly permitted Dr. Shah to re-test and adjust his technique during pre-trial testing. The actual negative pre-trial test results were never published alongside the published trial results indicating Dr. Shah's laboratory had the most sensitive technique to detect SV40 among the nine participating laboratories. PMID- 12552946 TI - Irinotecan and vinorelbine in patients with non-small cell lung cancer previously treated with platinum-based chemotherapy. A phase II study of the Hellenic Cooperative Oncology Group. AB - PURPOSE: To evaluate the efficacy and tolerability of irinotecan plus vinorelbine every 2 weeks in patients with advanced non-small cell lung cancer (NSCLC), previously treated with platinum-based chemotherapy. PATIENTS AND METHODS: Forty one patients with advanced NSCLC, refractory or resistant to platinum derivatives, were treated on an out-patient basis with irinotecan 150 mg/m2 intravenous (i.v.) and vinorelbine 25 mg/m2 on days 1 and 15. Chemotherapy was repeated every 4 weeks. The response was evaluated every two cycles. RESULTS: On an intent-to-treat analysis, 6 patients (14.6%) [95% confidence interval (CI) 5.57% to 29.17%] achieved partial response (PR), 15 (36.6%) stable disease (SD) and 20 (48.8%) progressive disease (PD). The median time to tumor progression (TTP) was 4.9 months (range 0.17-15.5 months), the median survival time was 7.8 months (range 0.9 to 19.6 months) and the 1-year survival rate was 37%. Symptomatic benefit response including improvement of performance status (PS), dyspnea, anorexia and fatigue, cessation of hemoptysis, fever and reduction of cough and pain was seen in 10 to 42% of patients. No patient experienced grade 3/4 anemia. Grade 3/4 thrombocytopenia occurred in 2 (5%) patients. Five patients (12%) developed grade 3/4 neutropenia and 5 (12%) had neutropenic fever that required hospitalization, but was successfully treated with antibiotics and G-CSF support. One patient (2%) developed grade 4 fatigue and was withdrawn. Other grade 3/4 adverse events included diarrhea (n = 3; 2 required hospitalization), alopecia (n = 5) and neurotoxicity (n = 1). Six patients required a dose reduction. CONCLUSION: The combination of irinotecan plus vinorelbine administered every 2 weeks demonstrated rather low activity in advanced NSCLC patients who had previously failed platinum-based chemotherapy, but it was well tolerated and was associated with increased 1-year survival rate and improvement in cancer related symptoms. PMID- 12552947 TI - Association of HPC2/ELAC2 polymorphism with prostate cancer risk in a Japanese population. AB - BACKGROUND: To investigate the relationship between two common variants (Ser217 and Ala541) of the HPC2/ELAC2 gene and prostate cancer risk in a Japanese population, we performed a case-control study. MATERIALS AND METHODS: Cases and controls consisted of 81 prostate cancer patients with a family history and 106 controls. Ser217 and Ala541 polymorphisms were analyzed by the restriction fragment length polymorphism method. RESULTS: In controls, 94.5% and 100.0% had wild-type Ser217Ser and Ala541Ala genotypes, respectively. 5.7% of the controls had the Leu217 genotype. No Thr541 genotype was observed in the controls. 92.6% and 97.5% of the cases had the Ser217Ser and Ala541Ala genotypes. No significant differences were observed in the genotypic frequencies between controls and cases. We stratified prostate cancer cases according to the pathological grade (low- or high-grade) or the clinical stage (localized or metastatic). There was no statistical difference between the genotypic frequencies between the groups. CONCLUSION: The present study suggested that the common variants in the HPC2/ELAC2 gene play a limited role in the risk of prostate cancer in the Japanese population. PMID- 12552948 TI - Which patients with advanced, proximal gastric cancer benefit from complete clearance of spleno-pancreatic lymph nodes? AB - BACKGROUND: The prognostic significance of dissecting spleno-pancreatic nodes remains unclear in patients with advanced proximal gastric cancer. MATERIALS AND METHODS: Data from a total of 104 patients (74 males and 30 females; age range, 21 to 76 years; mean, 56.0 years), who had undergone curative total gastrectomy combined with spleno-pancreatectomy for advanced proximal gastric cancer, were analyzed with respect to clinicopathological features and patient survival. RESULTS: Metastases to spleno-pancreatic nodes were found in 24 patients (23.1%). Tumor size > 40 mm (p = 0.0218), histologically-undifferentiated type (p = 0.0346) and both Japanese and TNM node-stages (p < 0.0001) were associated with metastases to these nodes. The 5-year survival rate of patients with a T2 tumor was 65.4% in patients with no metastases to the spleno-pancreatic nodes and 45.5% in patients with metastases to these nodes (p = 0.0699). No patients with a T3 tumor and metastases to the spleno-pancreatic nodes survived more than 4 years. CONCLUSION: Complete clearance of SP-nodes for patients with advanced proximal gastric cancer is beneficial for patients with a T2 tumor but not for patients with a T3 tumor. Metastases to these nodes appear to be rare in tumors less than 40 mm. Thus, this treatment should not be routinely performed in patients with proximal advanced gastric cancer. It should not be considered in patients with T3 tumors or with tumors < or = 40 mm. PMID- 12552949 TI - Phase II study of neoadjuvant chemotherapy in patients with surgically-proven, unresectable stage III non-small cell lung cancer. AB - BACKGROUND: The aim of this study was to assess whether patients with truly unresectable (bulky extracapsular N2, T4 for tracheobronchial angle or mediastinal organ invasion) stage III non-small cell lung cancer (NSCLC), as proven by cervical mediastinoscopy supplemented or not by left anterior mediastinoscopy and fiberoptic bronchoscopy or thoracotomy, could become resectable after induction cisplatin-containing chemotherapy. In addition, we studied the value of preoperatory magnetic resonance imaging (MRI), in evaluating the probability of achieving a radical resection after neoadjuvant chemotherapy. PATIENTS AND METHODS: Sixteen consecutive untreated stage III NSCLC patients were enrolled in the study. All the patients received two cycles of combination chemotherapy including cisplatin 100-120 mg/m2 intravenously (i.v.) days 1 and 22 and vinorelbine 30 mg/m2 i.v. days 1, 8, 15, 22 and 28 or vinblastine 5 mg/m2 i.v. days 1, 8, 15, 22 and 28. Thoracotomy was planned, after chemotherapy, for all non-progressive patients. No other treatment after surgery was devised following radical resection and patients with residual disease after surgery received standard post-operative radiotherapy. Response to treatment was evaluated by thorax CT and MRI two weeks after the last administration of chemotherapy. RESULTS: The overall complete resection rate was 38% (6 out of 16 patients). MRI was predictive of complete resectability in 80% of cases. In fact, 6 patients judged resectable were completely resected, 3 patients judged unresectable underwent only explorative thoracotomy or incomplete resection while MRI was unpredictive only in one case. The most important chemotherapy-related toxicity was hematological: eight patients (50%) had grades III-IV leukopenia. CONCLUSION: These results indicate that preoperative second generation cisplatin based chemotherapy can make resectable truly unresectable stage III NSCLC patients in only 38% of cases and that MRI is a reliable tool for assessment of radical resection probability after neoadjuvant chemotherapy. PMID- 12552950 TI - Thyroiditis and oncocytic carcinoma: incidental association? A case report. AB - BACKGROUND: Several studies have reported an association between Hashimoto's disease and thyroid carcinoma although the cause/effect relationship is still controversial. CASE REPORT: In this paper we report the case of a 38-year-old female who first presented with a clinical history of Hashimoto's thyroiditis. Ultrasound examination showed a diffuse thyroid irregularity more pronounced in the right lobe. FNAC (Fine Needle Aspiration Cytology) was performed in this area, and the cytological diagnosis was "Hashimoto's thyroiditis". The patient underwent clinical follow-up. Two years later, an ultrasound examination showed a nodular lump in the area previously aspirated. A new FNAC evidenced a Hurthle cell neoplasia. Therefore, the patient underwent surgery. The histological diagnosis was "Hurthle cell carcinoma". CONCLUSION: In the present case, the clinical and ultrasound history suggest the development of a malignant lesion strictly related to thyroiditis, as previously reported by other studies on papillary carcinoma. A cause-effect relationship between chronic lymphocytic thyroiditis and oncocytic neoplasia is still a matter of controversy. Therefore, patients suffering from chronic thyroiditis require a careful follow-up and, in case of nodules development, FNAC is recommended. PMID- 12552951 TI - Correlation of the genotypes for N-acetyltransferases 1 and 2 with double bladder and prostate cancers in a case-comparison study. AB - BACKGROUND: The arylamine N-acetyltransferases play a major role in the metabolic activation of carcinogenic amines that are present in cigarette smoke and a variety of other exogenous sources. The objective of this study was to determine the association of rapid or slow arylamine N-acetyltransferase (NAT) genotypes with smoking history and the risk for developing both bladder and prostate cancer. PATIENTS AND METHODS: The subjects analyzed were a case group of 17 double-cancer patients and 34 age-matched controls who had benign prostatic hypertrophy, but were asymptomatic for prostate or bladder cancers. Genotyping of NAT1 and NAT2 alleles was done by restriction fragment length polymorphism and/or sequencing of NAT genes amplified from genomic DNAs by the polymerase chain reaction (PCR). RESULTS: No significant correlation was found between NAT1 genotypes, double cancer, and smoking history. While NAT2-smoking interaction gave an odds ratio of only 1.70 (p = 0.117), a disproportionate number of cancer cases were genotypically rapid: 12 of 17 cancer cases vs. 13 of 34 controls (odds ratio 3.88; p = 0.040). CONCLUSION: Rapid NAT2 genotype correlated significantly with the development of double prostate-bladder cancer. PMID- 12552952 TI - Different pathways of 5-fluorouracil metabolism after continuous venous or bolus injection in patients with colon carcinoma: possible predictive value of thymidylate synthetase mRNA and ribonucleotide reductase for 5-fluorouracil sensitivity. AB - BACKGROUND: The mode of action of 5-fluorouracil (5-FU) was evaluated in patients with advanced colon cancer treated preoperatively with 5-FU. PATIENTS AND METHODS: After obtaining informed consent, 30 patients were randomized into three groups: untreated controls (n = 16), continuous intravenous infusion of 5-FU for 5 days at a dose of 320 mg/m2 (CIV group, n = 6), or bolus intravenous injection of 5-FU for 5 days at the same dose (bolus group, n = 8). Surgically-resected samples were analyzed by 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to detect sensitivity to 5-FU. Samples were also assayed for thymidylate synthetase (TS) activity, levels of 5-FU RNA (F-RNA), ribonucleotide reductase (RNR) activity, and TS mRNA levels expressed as the ratio of TS mRNA to beta-actin mRNA. RESULTS: While similar levels of TS inhibition were observed in the civ and bolus groups, F-RNA levels were significantly increased in the bolus group. The TS mRNA ratio was significantly higher in the 5-FU-resistant group than 5-FU-sensitive group. In the control group, the 5-FU-sensitive group showed higher RNR enzymatic activity compared to the 5-FU-resistant group. CONCLUSION: Our results suggested that the method of administration resulted in the induction of different 5-FU metabolic pathways. High TS mRNA and low RNR activity may be concerned with 5-FU-resistance in patients with advanced colon carcinoma. PMID- 12552953 TI - Prognostic value of laminin-5 in serous adenocarcinomas of the ovary. AB - BACKGROUND: The basement membrane is the first structural barrier that carcinoma cells must penetrate to pass from one compartment to another. In serous papillary cystadenocarcinoma of the ovary, Laminin alpha 1, beta 2, beta 3 and gamma 2 chains are present in about 50% of the neoplastic tissue. The aim of the present study was to evaluate the prognostic significance of the expression of Laminin in patients suffering from malignant serous surface epithelial-stromal tumors of the ovary. PATIENTS AND METHODS: Seventy tumor tissue specimens of consecutive patients treated by surgery between 1985 and 1995 for ovarian cancer, FIGO stages I-IV, were investigated by immunohistochemical methods. RESULTS: Sixteen tumors (22.9%) showed immunohistochemical staining of the cytoplasm and 55 (78.6%) staining of the basement membrane in neoplastic tissue. In multivariate analysis, basement membrane staining of Laminin showed a statistically significant influence with reduced overall survival (p = 0.029). CONCLUSION: The search for additional prognostic factors in ovarian cancer patients is still ongoing. The basement membrane staining of Laminin in malignant serous epithelial tumors of the ovary has been found to be valuable for further investigation. PMID- 12552954 TI - Altered expression of the receptor and ligand in the TGF beta signaling pathway in diffusely infiltrating colon carcinoma. AB - A diffusely infiltrating colorectal carcinoma (DICC) is a distinct clinicopathological entity demonstrating invasive growth with extensive matrix production and associated with an extremely poor prognosis. The factors producing these peculiar features, however, remain to be determined. Here we investigated the receptor and ligand of the transforming growth factor (TGF) beta signaling pathway in 32 DICCs and 60 common type colorectal carcinomas (CCCs). Immunohistochemical analysis revealed that the expression of TGF beta 1 was markedly increased in DICC, especially of the schirrous type, compared to CCC, whereas the expression of the TGF beta type II receptor (RII) was significantly decreased in DICC compared to CCC. The molecular basis for the reduced expression of RII was further investigated in 15 cases of DICC with a diminished RII expression. Two somatic mutations were found: one was a missense mutation in the kinase domain (Cys533Tyr), and the other was a G to C transition at the putative Sp1 binding site in the core promoter region. The latter mutation reduced the promoter activity, which may be due to a reduced affinity of the mutant sequence to the putative transcriptional regulator, which was neither Sp1 nor Sp3. No methylated cytosine was found at any CpG sites in the promoter region. These results suggested the presence of unidentified mechanisms for the diminished expression of RII in DICC cases. PMID- 12552955 TI - Neoadjuvant chemotherapy for intermediate/high-grade soft tissue sarcomas: five year results with epirubicin and ifosfamide. AB - BACKGROUND: Neoadjuvant chemotherapy for intermediate/high-grade soft tissue sarcomas (STS) may provide some advantages for facilitating the surgical resection of the tumor and for disease control. However its role as induction therapy before surgery should still be proved. PATIENTS AND METHODS: Twenty-one patients with intermediate/high-grade STS and tumor size > or = 5 cm were consecutively treated from 1997 to 2001 with neoadjuvant chemotherapy based on epirubicin 60 mg/m2/day on days 1 and 2 and ifosfamide 1.8 gr/m2/day on days 1 through 5 every three weeks. Evaluation of objective tumor response and toxicity were carried out according to WHO criteria. RESULTS: Nine partial responses were documented; stable disease in 11 patients, progressive disease in one patient. Apart from nine cases of grade 4 neutropenia, the treatment was generally well tolerated. Twelve patients underwent conservative and limb salvage surgery. CONCLUSION: This therapeutic approach seems to be effective in facilitating surgery. Neutropenia was the most significant toxicity but it was preventable or medically treatable with G-CSF support. PMID- 12552956 TI - Scrape and fine-needle aspiration cytology of elastofibroma. AB - BACKGROUND: Cytological findings of elastofibroma have rarely been reported and, to the best of our knowledge, have been illustrated in only 4 patients in the English language literature. MATERIALS AND METHODS: We retrospectively reviewed fine-needle aspiration biopsy (FNAB) and scrape smears in 5 cases of elastofibroma. RESULTS: All the lesions were typically located between the rib cage and the scapula. The cytological smears consisted of mature adipocytes, fibroblasts, collagen fibers, globular bodies and characteristic "braid-like" or "fern-like" structures, revealing degenerative elastic fibers. All the smears were specifically diagnosed as elastofibroma in combination with clinical and radiological findings. CONCLUSION: FNAB smears of elastofibromas may be hypocellular due to their densely fibrous nature and therefore diagnostic material may often be overlooked. Careful evaluation of the background of the smears coupled with full knowledge of the clinical and radiological findings, including magnetic resonance imaging, is required to establish the correct diagnosis. PMID- 12552958 TI - Outcome of patients with hepatitis C virus-related hepatocellular carcinoma occurring after interferon therapy. AB - To determine whether previous IFN therapy for chronic hepatitis C (HCV) infection influences the outcome of patients with hepatocellular carcinoma (HCC), 143 patients were enrolled in this study. Of 143 patients, 48 had received previous IFN therapy (IFN group) and the remaining 95 had not (untreated group). We estimated distant intrahepatic recurrence-free intervals and disease-specific survivals of the two groups by the Kaplan-Meier method and analyzed the difference by the log-rank test. Factors determining distant intrahepatic recurrence-free interval and disease-specific survival were studied by univariate and multivariate analysis using Cox proportional hazards regression model. The proportion of patients with single tumors was significantly higher in the IFN group (p = 0.026). The IFN group showed a significantly higher distant intrahepatic recurrence-free interval (p = 0.001) and disease-specific survival (p = 0.003). Moreover, multivariate analysis indicated that previous IFN therapy for chronic HCV infection was a significant independent factor for distant intrahepatic recurrence-free interval and disease-specific survival. These results indicate that previous IFN therapy reduces multicentric hepatocarcinogenesis of HCV-related HCC and improves the patients' survival. PMID- 12552957 TI - Simultaneous radiochemotherapy in pediatric patients with high-grade glioma: a phase I study. AB - BACKGROUND: Sandwich chemotherapy as first postoperative treatment might cause resistance to ensuing irradiation. Simultaneous radiochemotherapy might prevent this. PATIENTS AND METHODS: Twenty-nine children with high-grade glioma were treated from 1997-1999. The median age was 11.1 years. Tumor locations included: cerebral cortex 12, deep cerebral locations 11, cerebellum 3, spinal cord 3. Pons was excluded. Total (n = 9), subtotal (n = 3) and partial (n = 9) tumor resection or biopsy (n = 7) were followed by radiochemotherapy: fractionated radiation (1.8 Gyld up to 59.4 Gy) was given simultaneously with two cycles of chemotherapy (cisplatin 20 mg/m2/d x 5d, etoposide 100 mg/m2/d x 3d, and cisplatin, etoposide, ifosfamide: 1.5 g/m2/d). The data of a previous protocol (Cancer 89: 2131-7, 2000) in the same population were used as controls. RESULTS: Early progressive disease (PD 2 out of 25 patients) was significantly less frequent as compared to the control group (p = 0.031). The toxicity appeared tolerable. PMID- 12552959 TI - Hurthle cell tumors of the thyroid gland. Personal experience and review of literature. AB - BACKGROUND: Oncocytic cell neoplasm of the thyroid is currently recognized as a histological entity, but doubts still exist about its clinical and evolutionary categorization. Controversies concern occurrence and frequency of malignant forms, natural history and therapeutic strategies. MATERIALS AND METHODS: The authors report six cases of Hurthle cell tumor. Five cases were adenoma, one was carcinoma. Morpho-functional pre-operative evaluation and inter-operative histopathological test were performed in all patients. One patient underwent lobectomy (absence of unusual characteristics of the adenoma Hurthle cell) and five underwent total thyroidectomies (1 carcinoma). All patients were treated with suppressive hormonal therapy. RESULTS: No mortality and morbidity was recorded. All patients are undergoing follow-up (adenomas: average 64.2 months; carcinoma: 132 months) and none of them show recurrent symptoms. DISCUSSION: Hurthle cell tumors can be diversified in adenoma and carcinoma. Almost all reports classify oncocytic nodules as malignant when capsular and/or vascular invasion is present or when there is peri-thyroid tissue infiltration or lymphatic or hematic metastases. A clear differentiation between adenoma and carcinoma is determined by a histological test. Also an intra-operative histopathological analysis is sometimes unable to show minimal signs of invasion. Conflicting observations about the biological behaviour of Hurthle cell neoplasm lead to different therapeutic strategies. The authors believe lobectomy is the treatment of choice when a clear histological diagnosis of adenoma has been made. When carcinoma is diagnosed or when doubts exist after intraoperative histological test, the authors recommend total thyroidectomy followed by scintigraphic test and preventive radio-active therapy. All patients should be treated with suppressive hormonal therapy and undergo periodic check-ups. PMID- 12552960 TI - A phase-II study of liposomal doxorubicin and docetaxel in patients with advanced pancreatic cancer. AB - BACKGROUND: Both liposomal doxorubicin (LD) and docetaxel (D) have a broad range of activity against solid tumors, including advanced pancreatic cancer (APC), as single agents, while their combination has produced encouraging response rates in the treatment of several malignancies. We have conducted a Phase-II study in order to evaluate the tolerance and efficacy of their combination as front-line treatment in patients with APC. PATIENTS AND METHODS: Twenty-one chemotherapy naive patients with unresectable, locally-advanced or metastatic pancreatic cancer were enrolled. These included 16 males and 5 females with median age 66 years (range 57-80). Performance status (PS) was 0 (n = 10 pts), 1 (n = 7 pts) and 2 (n = 4 pts). D (80 mg/m2), and LD (30 mg/m2) were administered on day 1, every 3 weeks. RhG-CSF s.c. was given to all patients. At the time of analysis, all included patients were evaluated for toxicity and for response. RESULTS: A total of 92 cycles were administered (4.38 cycles/patient). Partial response was achieved in 6 patients, with a median duration of response of 3 months. Stable disease was observed in 7 patients and progressive disease in 8 patients. The median duration of survival was 10 months (95% CI, 6-14 months) and the actuarial 1-year survival rate was 33.33%. With regard to toxicity, grades 3,4 neutropenia occurred in 8 (38%) patients and grades 3,4 thrombocytopenia in 4 (19%) patients. Non-hematological toxicity was recorded in 15 (71%) patients: grades 3,4 diarrhea (3 pts, 14%), hypersensitivity reactions (3 pt, 14%), grade 2 neurotoxicity (6 pts, 29%) and palmar-plantar erythrodysesthesia (9 pts, 43%). CONCLUSION: The doxorubicin and docetaxel combination was well-tolerated by these poor prognosis patients. Although both drugs have a marginal activity in pancreatic cancer, most patients experienced significant clinical improvement, with acceptable toxicity. PMID- 12552961 TI - The rational development of capecitabine from the laboratory to the clinic. AB - Although the anti-neoplastic activity of 5-Fluorouracil (5-FU) is improved by continuous infusion or biochemical modulation, the need for in-dwelling central venous catheters and toxicity have proved to be major impediments. Capecitabine (Xeloda, N4-pentyloxycarbonyl-5'-deoxy-5-fluoro-cytidine), an oral fluoropyrimidine, has been synthesized in the laboratory as an inactive precursor that passes intact through the intestinal mucosa and is sequentially converted to 5'-deoxy-5-fluorocytidine (5'-DFCR), 5'-deoxy-5-fluorouridine (5'-DFUR) and finally 5-FU in the liver and tumour tissues selectively. Preclinical studies provided evidence of preferential conversion of inactive 5'-DFUR to active 5-FU in solid tumours due to the relative overexpression of the final anabolising enzyme, thymidine phosphorylase (TP), in neoplastic tissues more than in normal counterparts. Phase I/II studies exploring toxicity and the appropriate dosing resulted in the evaluation of the intermittent schedule (2510 mg/m2/day for 14 days every 3 weeks) in subsequent trials. Two large phase III studies in patients with metastatic colon cancer established at least equivalent efficacy and improved tolerability for capecitabine compared to the Mayo Clinic bolus 5 FU/folinic acid regimen. Phase II studies established remarkable activity in women with heavily pretreated metastatic breast cancer. Moreover the combination with a taxane yielded a unique survival benefit compared to the previous gold standard of taxane monotherapy in a phase III trial of women with anthracycline resistant breast cancer. The commonest side-effects are hand and foot syndrome, diarrhoea and stomatitis with serious adverse events occurring in a minority of patients. Myelosuppression was minimal or absent. Toxic manifestations are easily managed with a significant reduction in the frequency of hospitalizations and medical resource use, as shown in appropriate studies. Capecitabine is expected to find a role in the treatment of other tumour types as well as adjuvant administration. It represents an advance in modern drug development, stressing the current shift towards rational development of new agents and home-based outpatient regimens. PMID- 12552962 TI - Tumors in organ transplant recipients may give clues to their control by immunity. AB - Tumors in solid organ transplant recipients are an important complication of surgery. They can be due to the recurrence of malignancy existing in the recipient prior to transplantation, tumors of donor origin transmitted inadvertently and de novo malignancies. These patients constitute a sort of experimental group in whom the normal immune control of the host has been weakened and can provide valuable information. The reported distribution of the tumors developed in these patients may indicate those whose development is controlled by the immune system. Some of the reported data has been unexpected. For example, patients grafted to treat a primary cancer or in whom an asymptomatic tumor was discovered at the time of transplantation rarely have a recurrence. Many of these were skin tumors, but why the SCC/BCC incidence ratio is far from 1 is unclear. Melanomas are not more frequent among immunosuppressed grafted patients, in spite of the fact that they have specific antigens which could be targets for immune and tumor growth control. Some tumors regress and disappear when the immunosuppression regimen is withdrawn. Tumor types rarely observed in grafted patients are thus immune-insensitive and would not normally regress due to immunotherapy. PMID- 12552963 TI - Independent antitumor spectrum of UCN-01 (7-hydroxystaurosporine) against gastric and colorectal cancers as detected by MTT assay. AB - To evaluate the clinical usefulness of 7-hydroxystaurosporine (UCN-01), we compared the antitumor spectrum of UCN-01 with those of conventional antitumor agents against 40 fresh gastric and 40 fresh colorectal cancer specimens using the MTT assay. At a cut-off concentration of 30 micrograms/ml, UCN-01 showed a higher efficacy rate than mitomycin C (MMC), cisplatin, and 5-fluorouracil (5-FU) against both gastric and colorectal cancers. With respect to the gastric cancer specimens, the antitumor spectrum of UCN-01 was independent from those of the other agents, while the patterns of antitumor effects of the conventional agents all correlated significantly with each other. For the colorectal cancer specimens, the pattern of UCN-01-sensitivity did not correlate with the patterns for 5-FU or MMC. In conclusion, UCN-01 may be useful for clinical application against gastric and colorectal cancer due to its different antitumor spectrum from conventionally available agents. PMID- 12552964 TI - Axillary metastasis as the first manifestation of occult breast cancer in a man: a case report. AB - Breast cancer is extremely rare in men, accounting for less than 1% of all breast cancers. We describe a male patient with occult breast cancer that was confirmed immunohistochemically. To our best knowledge, this is the first case presenting axillary and supraclavicular metastases as the first manifestation of occult cancer of the male breast. PMID- 12552965 TI - Correlation between BRCA1 expression and apoptosis-related biological parameters in sporadic breast carcinomas. AB - BRCA1 is a tumor suppressor gene that is responsible for hereditary breast and ovarian cancer syndrome. Increased evidence suggests that BRCA1 protein is involved in mammary carcinogenesis in sporadic and hereditary forms. Recent experimental results suggest that BRCA1 plays a role in the regulation of apoptosis. In order to test whether the analysis of human tumors would provide data supporting this hypothesis in sporadic breast carcinomas, we have investigated the relationship between BRCA1 and apoptosis-related genes. Immunohistochemical analysis was performed to determine BRCA1 and the apoptosis related proteins bcl-2, Bax and p53 in paraffin-embedded tissues of 156 sporadic invasive ductal carcinomas. BRCA1 expression was positively-correlated with Bcl-2 expression (p = 0.0008), but no relationship between BRCA1 expression and Bax or p53 expression could be established. In addition, loss of BRCA1 expression was also related to poor tumor differentiation and lymph node metastasis. Our study shows that bcl-2 might be one of the target genes involved in the oncogenesis related to BRCA1. Loss of BRCA1 may contribute to tumor development in breast carcinomas, which may be independent of the p53 tumor suppressor. PMID- 12552966 TI - A phase II study of weekly methotrexate, cisplatin, and 24-hour infusion of high dose 5-fluorouracil and leucovorin (MP-HDFL) in patients with metastatic and recurrent esophageal cancer-improving toxicity profile by infusional schedule and double biochemical modulation of 5-fluorouracil. AB - BACKGROUND: Patients with advanced esophageal cancer are frequently not indicated for intensive chemotherapy due to aero-digestive tract complications. In order to improve the therapeutic index of systemic chemotherapy for these patients, we designed a weekly MP-HDFL protocol, which took advantage of the low toxicity of the infusional dosing schedule for cisplatin and 5-fluorouracil (5-FU), and double biochemical modulation of 5-FU by methotrexate and leucovorin. PATIENTS AND METHODS: Between April 1997 and April 2001, a total of 26 patients (M:F = 22:4) with a median age of 59.5 (range: 43-81) years were enrolled into this phase II clinical trial prospectively. There were 12 patients with de novo metastatic disease, 13 with recurrent disease (including 7 relapsed from previous concurrent chemoradiotherapy incorporating cisplatin and 5-FU), and one with locally advanced disease. The weekly MP-HDFL protocol was composed of methotrexate 40 mg/m2, i.v., on days 1, 8, 15; cisplatin 35 mg/m2, 24-hour IVF, on days 2, 9, 16; and 5-FU 2,600 mg/m2 and leucovorin 300 mg/m2, 24-hour IVF, on days 2, 9, 16; repeated every 28 days. RESULTS: A total of 97 courses were given with an average of 3.7 courses per patient (range: 1-19). All of the patients were assessable for evaluation of toxicity. Grades 3/4 leukopenia and thrombocytopenia were noted in 13 (14%) and 17 (18%) of the treatment courses, respectively. There were 3 episodes of febrile neutropenia in 2 patients, and lethal sepsis in one patient. The Grades 3/4 non-hematological toxicities were infrequent. Among 25 patients with recurrent or metastatic disease, 7 had partial response and 12 had stable disease. The response rate for the intent- to treat group was 28% (95% C.I.:12%-49%). The median survival was 5 months (range: 1-28 months). CONCLUSION: Weekly MP-HDFL combination chemotherapy is associated with mild and acceptable toxicity, and has modest palliative activity in a portion of advanced esophageal cancer patients. However, a more active and better tolerated anticancer drug combination is needed to improve the outcome of these patients. PMID- 12552967 TI - Chemotherapy for small cell lung cancer in patients over 80 years old. AB - BACKGROUND: Elderly cancer patients sometimes are not considered candidates for standard chemotherapy. We evaluated the efficacy of chemotherapy for small cell lung cancer (SCLC) in patients aged over 80 years old. PATIENTS AND METHODS: We retrospectively analyzed the records of 10 SCLC patients over 80 years old. Six of them had received carboplatin and etoposide intravenously, two had etoposide orally, and two had only supportive care. RESULTS: Seven of the 8 patients treated with chemotherapy responded partially and the median survival time was 281 days. Despite the toxicity of the chemotherapy, the performance status was improved in 5 out of 8 treated patients and the cancer-related symptom was relieved in all treated patients for at least one month during and/or after chemotherapy. CONCLUSION: Chemotherapy is indicated for SCLC patients even aged over 80 years. The quality of life is improved by chemotherapy, although the survival time may not be prolonged significantly. PMID- 12552968 TI - Phase II study of mitomycin C, cisplatin and 5-fluorouracil for advanced and recurrent gastric cancer. AB - In this study, we assessed the efficacy and feasibility of a 5-FU, MMC and cisplatin combination in patients with advanced or recurrent gastric cancer. 5-FU was administered at a dose of 360 mg/m2 on days 1 through 5 and days 8 through 12. CDDP was administered at a dose of 7 mg/m2 on days 1 through 5 and days 8 through 12. MMC was given at a dose of 13 mg/m2 on day 1. Twenty-seven patients with non-resectable or recurrent gastric cancer were entered. The most common toxicity was leukopenia. Nausea/vomiting was generally mild and no patient suffered severe diarrhea, mucositis or renal insufficiency. While a complete response was not observed, 13 patients showed a PR giving an overall response rate of 48.1% (95%CI, 28.0 to 68.3%). Our regimen may have advantages in terms of reduced toxicity with moderate efficacy that is comparable with results using the ECF regimen. PMID- 12552969 TI - Increasing incidence of cholangiocarcinoma in Crete 1992-2000. AB - BACKGROUND: There have been recent reports of an increased incidence of cholangiocarcinoma. PATIENTS AND METHODS: Patients with cholangiocarcinoma, resident on the island of Crete, diagnosed between 1992 and 2000, were studied. Case ascertainment was enhanced by collaboration of the pathology departments and all gastroenterologists of Crete in providing relevant data. A specially designed patient protocol was used. Crude incidence rates are presented. RESULTS: During the 9-year period 7 patients were diagnosed with cholangiocarcinoma of the intrahepatic ducts, 17 with Klatskin and 16 patients with extrahepatic ducts carcinoma. A steady incidence increase of cholangiocarcinoma, irrespective of location, was shown. The estimated incidence rate per three-year period progressively increased from 0.998 to 2.329 and 3.327 per 100,000 for the periods 92-94, 95-97 and 98-2000, respectively. Median survival time was 8.5 months. No significant difference was seen between survival according to the location of cholangiocarcinoma. CONCLUSION: A steady incidence increase of cholangiocarcinoma cases in Crete during the time-period 1992-2000 was shown. PMID- 12552970 TI - Solitary splenic recurrence of epithelial ovarian cancer: a case report and review. AB - The metastatic involvement of the spleen in epithelial ovarian cancer is rare and usually reflects late disseminated disease. Isolated parenchymal metastasis in the spleen is an extremely unusual event, in fact in most cases in medical literature, the spleen was involved as part of diffuse peritoneal carcinomatosis. We report a rare case of epithelial ovarian carcinoma, which recurred in the splenic parenchyma 15 years after initial treatment. The patient underwent a splenectomy followed by six cycles of platinum-based poly-chemotherapy. Currently, 8 years after splenectomy, the patient has no evidence of relapsed disease. In the solitary splenic recurrence of epithelial ovarian cancer our case suggests that accurate surgical resection of the spleen, followed by platinum based chemotherapy, is correlated with a prolonged survival. PMID- 12552971 TI - Glutathione S-transferase (GST) polymorphisms as risk factors for cancer in a highly homogeneous population from southern Italy. AB - Glutathione S-Transferase (GST) is a phase II enzyme and catalyses reactions between glutathione and a variety of electrophilic compounds, including some environmental carcinogens. In man, at least 20 isoenzymatic forms of GST have been identified and many of them show genetically-based individual variability of enzyme activity. The GSTM1 and GSTT1 isoenzymes display several polymorphisms, including a homozygotic deletion, which have been associated with an increased risk for developing neoplastic diseases. There is geographical and ethnic variation in genotype frequencies for both genes. The available data suggest that cancer incidence varies amongst Italian regions, being higher in Northern that in Southern areas, though it is unknown whether this phenomenon is to be attributed to genetic and/or environmental factors. We performed a case-control study to evaluate the GSTM1 and GSTT1 polymorphisms in a series of cancer patients in Basilicata, a Southern Italian region, and in corresponding controls. The results obtained demonstrate that the occurrence of GST polymorphisms in the Basilicata population is not different from other Italian regions and suggest that the population attributable risk associated with these genotypes may be quite high. GSTM1 homozygous null genotype was associated with an increasing risk of cancer, especially in females. The strongest association was with colon and breast cancers. For the GSTT1 gene, the results obtained were suggestive of a decreased risk of cancer associated with the null genotype. Thus, similar studies on these and other susceptibility genes are warranted since they can help to identify susceptible subgroups of people who can be targeted for cancer prevention. PMID- 12552972 TI - Thymidylate synthase and p53 expression in primary tumor do not predict chemotherapy outcome in metastatic colorectal carcinoma. AB - INTRODUCTION: Thymidylate synthase (TS) and p53 expression have been reported to predict the results of palliative chemotherapy in advanced colorectal carcinoma (ACRC), but the knowledge is still limited and partly conflicting. PATIENTS AND METHODS: One hundred and twenty-two patients with ACRC were treated with 5 fluorouracil (5-FU)-based therapy at the University Hospital in Uppsala, Sweden, in four different randomised phase III studies between 1989 and 1997. Fifty-nine (48%) of the patients were initially diagnosed with advanced disease. There were 32 (26%) complete or partial radiological responders. The paraffin-embedded tumours at primary diagnosis were retrospectively analysed with immunohistochemical techniques (IHC) for TS using the specific monoclonal antibody, TS 106, and for p53 with the mouse monoclonal antibody DO-7. RESULTS: All analyses were independently reviewed. High TS values were seen in 78% and p53 positivity in 60% of the tumours. None of the markers predicted the outcome of the later palliative treatment, either in terms of an objective response or survival. However, for the subgroup who initially had curative resection (Dukes' A-C), TS expression had prognostic information and significantly predicted time to recurrence (median for low TS tumours 30 months and for high TS tumours 11 months, p = 0.001). CONCLUSION: Immunohistochemical investigation of TS and p53 of the primary tumour is not useful to predict the outcome of palliative chemotherapy in ACRC. Instead, TS can be regarded as a marker for prediction of time to recurrence. PMID- 12552973 TI - The relevance of weight loss for survival and quality of life in patients with advanced gastrointestinal cancer treated with palliative chemotherapy. AB - BACKGROUND: Chemotherapy is frequently used for palliation in patients with advanced gastrointestinal (GI) cancer and improvements in quality of life (QoL) and survival are documented. Several pre-treatment factors, particularly poor performance status, influence the outcomes. The relative importance of nutrition associated factors is less well known. PATIENTS AND METHODS: Data from 152 patients treated within four randomised studies were analysed to explore the relations between weight loss, nutrition-associated parameters, performance status, response to chemotherapy, global QoL and survival. RESULTS: Patients with weight loss, poor performance, higher frequency of nutritional problems and low haemoglobin levels had a lower probability of responding to treatment. Global QoL was decreased in patients having weight loss as well as problems with pain or fatigue. Diagnosis had the strongest association to survival. In a multivariate analysis poor performance and weight loss were also independently related to survival. CONCLUSION: Weight loss and several nutrition-associated problems were associated with lower QoL, less likelihood to respond to chemotherapy and poor survival. PMID- 12552974 TI - Tissue polypeptide specific antigen (TPS) and carbohydrate antigen 125 (CA-125) in the early prediction of recurrent ovarian cancer. AB - It is very important to establish a tumor marker combination with strong lead time effects in early detection of recurrent ovarian cancer. This retrospective study included 32 patients with recurrent epithelial ovarian cancer after primary therapy. The serum levels of tissue polypeptide specific antigen (TPS) and carbohydrate antigen 125 (CA-125) were followed-up. Normal upper limits of serum levels of 78.5 U/l for TPS and 35 U/ml for CA-125 were selected according to the 95th percentile of serum concentrations measured in healthy control patients. When compared with other follow-up modalities, TPS and CA-125 appeared lead-time effective with early diagnosis of recurrent ovarian cancer in 18 and 16 patients, respectively. This difference was not statistically significant. The combination of TPS and CA-125 provided lead-time effects in 24 patients. Our data indicate that the combination of TPS and CA-125 is a potential tool in the early prediction of recurrent ovarian cancer. PMID- 12552975 TI - Is tumor size a prognostic indicator for gastric carcinoma? AB - BACKGROUND: Retrospective studies using multivariate analysis of clinical and pathological features in gastric cancer have identified a number of high-risk, independent prognostic factors. In these clinical features, tumor size can be measured easily before or during the operation without the requirement of any special equipment, but its prognostic value in patients with gastric cancer is unclear. The aim of this study was to review the experience at our institution of gastric adenocarcinoma to determine the influence of tumor size on outcome. PATIENTS AND METHODS: Between January 1985 and December 1995, 697 patients with gastric adenocarcinoma underwent resection of the stomach at the Department of Surgery, Sendai National Hospital, Japan. Data on age, gender, tumor location, structure and size, evidence of local invasion, and type of operation performed for each patient were obtained. The sizes of tumors were derived from measurements made in fresh resected specimens. The patients were divided into three groups: 102 patients with tumors of less than 2 cm in diameter, 392 patients with tumors of 2-7 cm in diameter, and 203 patients with tumors of more than 7 cm in diameter. RESULTS: In these three groups, there were statistical differences in tumor location, macroscopic type, depth of invasion, lymph node metastasis, histological type, lymphatic invasion, vascular invasion and cancer stromal relationship. Patients with larger tumors had more invasion into the gastric wall in terms of depth of invasion and more frequent lymph node metastasis than did patients with smaller tumors. Histologically, diffuse, scirrhous-type was more common in the larger tumor group. The frequency of lymphatic and vascular permeation in the larger tumor group was higher than that in the other groups. The 5-year survival rates according to tumor size were 94.3% in cases of tumors of less than 2 cm, 75.1% in cases of tumors of 2-7 cm, and 26.3% in cases of tumors of more than 7 cm. Multivariate analysis revealed that the prognosis of gastric cancer patients was affected most by depth of invasion, followed by lymph node metastasis and tumor location. Tumor size is not an independent prognostic factor. CONCLUSION: In conclusion, according to the results of univariate analysis, tumor size is clinically a predictor of survival of patients with gastric cancer. In multivariate analysis, however, it is not an independent factor, and the presence of lymph node metastasis, depth of invasion and tumor location are more important than tumor size. PMID- 12552976 TI - Predictive value of CD8highCD57+ lymphocyte subset in interferon therapy of patients with renal cell carcinoma. AB - BACKGROUND: The aim of this retrospective study was to evaluate the significance of pre-treatment levels of peripheral blood lymphocyte subsets for survival of renal cell carcinoma (RCC) patients treated with interferon-a2b (IFN). PATIENTS AND METHODS: CD3+, CD19+, CD16&56+, CD4+, CD8+, CD4+CD45ROhigh and CD8highCD57+ lymphocyte subsets in peripheral blood of 85 advanced RCC patients were determined using flow cytometry. The survival of IFN-treated and non-treated patients was analyzed by gradually testing different cut-off levels of each lymphocyte subset. RESULTS: Advanced RCC patients with > or = 30% CD8highCD57+ lymphocytes in the CD8+ subset had shorter survival compared to patients with < 30% CD8highCD57+ lymphocytes in the CD8+ subset (p = 0.01). Treatment with IFN increased overall survival only in the former subgroup of RCC patients (p = 0.02). CONCLUSION: The present study suggests that the percentage of CD8highCD57+ lymphocytes in the CD8+ subset may have a prognostic significance for advanced RCC patients and may have a predictive value in patient selection for survival benefit due to treatment with IFN. PMID- 12552977 TI - Reimplantation of autoclaved or irradiated cortical bones invaded by soft tissue sarcomas. AB - BACKGROUND: Bone defects after excision of malignant bone and soft tissue tumors are usually reconstructed by autogenous bone grafting or allografting. PATIENTS AND METHODS: We treated 2 patients who had soft tissue sarcomas with bone involvement, the first by excision and reimplantation of the hemicortical autogenous cortical bone graft after autoclaving and the second with intraoperative irradiation. RESULTS: In both patients, postoperative follow-up radiographic studies showed incorporation of the implanted bones into the host bones within 12 months. Biopsy of the implanted bone performed 3 years after surgery showed many viable osteocytes, and blood vessel invasion into Harversian canals. No complications regarding the reconstructive techniques were observed during the postoperative period. CONCLUSION: The reimplantation of autogenous cortical bones after autoclaving or irradiation is a useful reconstructive method for partial bone defects in soft tissue tumor surgery. PMID- 12552978 TI - Characteristics of clear cells and Toker cells in the epidermis of underlying nipple duct adenoma. AB - Clinically, it is difficult to differentiate between nipple duct adenomas (NDAs) and Paget's disease of the nipple. These lesions share similar morphological and histological characteristics. Clear cell types present in NDA, epidermal clear cells (ECC) and Toker cells (TC), share immunoreactive similarities to Paget cells which can lead to confusion in classification. The aim of this study was to obtain information on the characteristics and histogenesis of ECC and TC, to distinguish these cells from Paget cells. Ten nipple epidermal with NDA were compared to 25 histologically normal nipples. Samples were analyzed for cytokeratins (CKs) 7, 8 and 18, carcinoembryonic antigen (CEA), c-erbB-2/HER2 expression and human papillomavirus (HPV-) DNA. In 13 out of 25 normal nipples the staining sequence demonstrated that ECC and TC cell types are immunoreactive with CKs 7, 8 and 18 in the basal region of the epidermis. In contrast, aggregated CKs 7, 8 and 18-immunoreactive ECC and TC were identified in the epidermal of 8 of the 10 NDA cases. In 2 cases, TC were continuous with the underlying NDA, suggesting that TC might be of ductal origin and migrate through the galactophorous ostia. In NDAs and 25 histologically normal nipples, ECC and TC were negative for CEA, c-erbB-2/HER2 and HPV-DNA. ECC and TC, normally present in the nipple epidermis, may proliferate and form aggregates in the presence of an underlying NDA. These cells show immunoreactivity for CKs 7, 8 and 18 but are negative for c-erbB-2/HER2, CEA and HPV-DNA and should not lead to the mistaken diagnosis of Paget's disease. PMID- 12552979 TI - Evaluation of infectious complications and immune recovery following high-dose chemotherapy (HDC) and autologous peripheral blood progenitor cell transplantation (PBPC-T) in 148 breast cancer patients. AB - BACKGROUND AND OBJECTIVES: High-dose chemotherapy (HDC) with autologous PBPC-T has been reported to be effective in hematological and in selected solid malignancies. In this setting, infectious complications represent a relevant cause of morbidity. PATIENTS AND METHODS: To ascertain the incidence, types and factors influencing the development of early and late infections, we retrospectively analyzed 148 consecutive breast cancer (BC) patients receiving HDC and PBPC-T, both for primary high-risk BC (pBC) and metastatic disease (mBC). RESULTS: Early infection strongly associated with the occurrence of grade 4 mucositis (p < 0.001), was documented in 28 patients (19%). Late re-activation of varicella zoster virus (VZV) occurred in 14 patients (9%); an inverse correlation between the VZV re-activation and the total amount of T-cells transferred with the graft was observed. Evaluation of immune reconstitution, carried out in 10 out of 148 patients, showed a long-lasting CD+ T-cells depression (> 2 year), mainly involving the naive CD4+ T-cell subset. Conversely, the analysis of the frequency of proliferating T-lymphocyte precursors, specific for antigens expressed by 3 different widespread pathogens, demonstrated that, notwithstanding the delayed recovery of CD4+ cells, many T-lymphocyte functions were within normal range 1 year after PBPC-T. CONCLUSION: Altogether these results show that severe mucositis is associated with early bacterial infections and the infusion of large numbers of T-cells plays a role in controlling late VZV reactivation. PMID- 12552980 TI - Surgical treatment of hepatocellular carcinoma in elderly cirrhotic patients: personal experience. AB - BACKGROUND: Due to the longer life expectancy of the world's population, the number of elderly cirrhotic patients undergoing surgery for hepatocellular carcinoma (HCC) is increasing. Our study evaluates the benefits of hepatic resections for HCC in cirrhotic patients aged over 65, analysing the early and long-term surgical results. PATIENTS AND METHODS: We retrospectively considered a series of 46 patients receiving hepatic resection for HCC. The clinicopathological data and surgical outcome of 14 (30.4%) patients aged 65 or older (group I) were evaluated and compared to the 32 (69.6%) younger than 65 (group II). RESULTS: No operative mortality was recorded in either group. The hospital mortality rate was 7.1% (1 out of 14) in group I and 9.4% (3 out of 32) in group II (p = 1.00). Hospital morbidity did not differ significantly in the two groups (21.4% vs 34.4%; p = 0.50). At follow-up (median 34 months, interquartile range: 12-63) 3 patients from group I (21.4%) and 16 patients from group II (50%) experienced tumor recurrence (p = 0.14). The five-year disease free survival rate for group I and group II was 71.4% vs 28.2%, respectively (p = 0.05). The overall 5-year survival rate for group I and group II was, respectively, 77.4% vs 41.8%, (p = 0.3). CONCLUSION: Elderly cirrhotic patients with HCC can benefit from hepatic resection as well as younger patients; age by itself should not be considered a contraindication to surgery. PMID- 12552981 TI - Prognostic value of tissue-polypeptide specific antigen (TPS) in bladder cancer. AB - BACKGROUND: To evaluate, by means of a prospective study, the usefulness of tissue polypeptide specific antigen (TPS) as a tumor marker in follow-up and prognosis of bladder cancer. PATIENTS AND METHODS: A total of 438 subjects were included in the study and divided into three groups. The first group (n = 216) had active bladder cancer disease, the second group (n = 168) known bladder tumor disease but with no recurrence at the time of study, while the third group (n = 54) consisted of healthy volunteers. TPS in serum was measured using an EIA-kit, a solid phase two-site immunosorbent assay with polyclonal antibodies against cytokeratin 18. RESULTS: Serum TPS was significantly higher in patients with bladder tumors (p < 0.05). There was a significant correlation between TPS and stage (p < 0.05), presence of metastatic lymph nodes (p < 0.01), metastasis (p < 0.01), urinary cytology (p < 0.05), tumor shape (p < 0.01) and tumor size (p < 0.05). Grade, number of tumors, relapses and recurrences were not significantly correlated with serum TPS. With a TPS concentration of 100 U/L as the cut-off point, the sensitivity was 37% and the specificity 77%. With a 95% specificity, the sensitivity was 9%. CONCLUSION: Serum TPS is a marker for bladder carcinoma correlated with stage, urine cytology, tumor shape and size. Its clinical usefulness is not, however, established and it does not appear to be of use in the follow-up of patients with bladder tumor disease. PMID- 12552982 TI - Kinetics of postoperative serum vascular endothelial growth factor (VEGF)--can it be used to predict curative resections in colorectal cancer? AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is considered to be probably the most important and final mediator of neovascularisation. We have shown that VEGF can predict stage in colorectal cancer (Clin Cancer Res 1998, 17). This study was conducted to study the kinetics of serum VEGF after colorectal resections over ten postoperative days. PATIENTS AND METHODS: The study comprised 154 healthy controls and 108 colorectal resections (79 curative, 15 palliative and 14 benign). Samples were collected at 4-6 hours, days 1,3,5,7 and 10 post surgery. RESULTS: Six-hour levels were significantly lower in the curative group (p < 0.0005) but not in the benign and palliative groups (p = 0.27 and 0.3, respectively). Sensitivity and specificity at 20% cut-off fall in VEGF gives 83.5% sensitivity with 80% specificity in predicting curative resection. CONCLUSION: Early postoperative serum VEGF levels show significant fall and may help to identify the oncological status of colorectal cancer resections. PMID- 12552983 TI - Sequential high-dose chemotherapy with autologous stem cell support in patients with limited-stage small cell lung cancer. AB - To determine the feasibility and safety of multiple sequential courses of high dose chemotherapy with peripheral blood progenitor cells (PBSCs) followed by thoracic irradiation (54 Gy) and prophylactic cranial irradiation (PCI 30 Gy) in patients with limited disease (LD) small cell lung cancer (SCLC), six patients received one cycle of conventional-dose chemotherapy followed by four cycles of high-dose chemotherapy with stem cell support. Neither significant adverse side effects nor treatment mortality were observed. CR and almost CR (> 90%) was obtained in five out of six patients while one patient had a partial response (PR). The median survival was 21.3 months (range 8-34). However, the benefit of the high CR rate and prolonged survival did not result in cure even in these favorable circumstances, because all the patients died of recurrence in the cerebral nervous system. According to our data we stopped this chemotherapy regime because it was not able to prevent cerebral disease. PMID- 12552984 TI - Efficacy of prophylactic anti-diarrhoeal treatment in patients receiving Campto for advanced colorectal cancer. AB - This study assessed the efficacy of combined prophylactic and curative anti diarrhoeal medication in advanced colorectal patients treated by irinotecan. Thirty-four pre-treated eligible patients were evaluated. There were 44% women, the median age was 65 and 38% of the patients had a 0 performance status. The patients received sucralfate(4g/d) and nifuroxazide(600 mg/d) prophylactic treatment on days 0-7. In the case of severe diarrhoea, preventive treatment was replaced by loperamide(12 mg/d) and diosmectite (9 g/d). Grade 3 delayed diarrhoea occurred in 18% of patients (90% CI: [9.5-28.9]) and 4.6% of cycles. No grade 4 delayed diarrhoea was observed. Twenty-nine patients (85%) received the preventive treatment at cycle 1, while 14% (90% CI: [6.2-25.7]) experienced grade 3 delayed diarrhoea in 3.7% of cycles for a median 4.5 days. The objective response rate was 8% (90% CI [1.4-23.1]) among the 25 assessable patients. Preventive combined treatment is effective in reducing the incidence of severe delayed diarrhoea, and it should be proposed to patients treated with mono therapy Campto(r) and evaluated in poly-chemotherapy protocols. PMID- 12552985 TI - Is therapeutic conization sufficient to eliminate a high-risk HPV infection of the uterine cervix? A clinicopathological analysis. AB - BACKGROUND: High-risk human papillomavirus infection plays a predominant role in the pathogenesis of preinvasive and invasive cervical cancer. One of the recognized treatments of cervical intraepithelial neoplasia is conization. The aim of this study was to evaluate if cold-knife conization is sufficient to eliminate cervical intraepithelial neoplasia and the associated high-risk HPV infection. PATIENTS AND METHODS: Thirty-seven high-risk HPV-positive women who underwent cold-knife conization entered this study. The cervical sampling for HPV DNA was performed using the Digene cervical sampler. Smears were taken immediately before and 3 months after conization and the patients were followed up for 2 years. RESULTS: High-risk HPV was identified in all 37 patients before conization. In 4 out of 37 patients a coincidence of low/intermediate and high risk HPV types was present. A CIN II was detected in 5 out of 37, a CIN III in 25 out of 37 and a carcinoma in situ in 7 out of 37 cases. Follow-up at three months revealed that HPV was eradicated by conization in 73%. Patients with persistent HPV infection tended to be older compared to patients with eliminated HPV infection (mean: 34 vs. 36 years; p = 0.25) and showed a higher rate of severe dysplasia (p = 0.07). A high HPV prevalence among patients with positive resection margins and/or recurrence disease was detected (83% and 100%, respectively). A statistically significant higher rate of positive margins and recurrences was observed in patients with persistent compared to patients with eliminated HPV infection (50% vs. 4%. p = 0.001 and 30% vs. 0%, p = 0.003). CONCLUSION: The data of the present study demonstrated that a high-risk HPV infection is successfully eliminated by conization in most cases. A high HPV prevalence in patients who had positive cone margins and/or disease recurrence was observed. Patients with persisting HPV infection after conization show statistically significant higher rates of positive resection margins and are at increased risk of disease recurrence. HPV testing seems to be, therefore, a valuable tool to monitor the therapeutic results of conization and to discriminate patients who have a higher risk of disease recurrence. PMID- 12552986 TI - Prognostic value of DNA ploidy status in patients with rectal cancer. AB - BACKGROUND: A retrospective study was performed to measure the prognostic value of DNA ploidy status and proliferative index (PI) for survival in patients with rectal cancer. PATIENTS AND METHODS: Fifty-two patients underwent curative surgery for rectal carcinoma. Ten tumors were in Stage I, 25 cancers were in Stage II, and 17 of them were in Stage III. Using flow cytometry the nuclear DNA content of the tumor cells was measured. RESULTS: There were 25 DNA diploid and 27 DNA aneuploid carcinomas. Aneuploid DNA content did not show higher occurrence in advanced tumors. The mean survival was 59 months in the case of DNA diploid carcinoma, while it was 47 months in the case of DNA aneuploid cancer. The mean PI of the DNA diploid cancers was 8%. The PI of DNA aneuploid tumors was 22%. High PI (PI > 10%) was observed in 32 carcinomas while low PI (PI < 10%) occurred in 20 cases. Patients with aneuploid DNA content and high PI had significantly worse survival compared to patients with diploid DNA content while low PI. Locoregional and distant metastases occurred more frequently in patients with aneuploid tumor. By univariate analysis, tumor size, lymph node involvement, DNA ploidy status and PI all correlated with prognosis. However, multivariate analysis showed that TNM stage and PI were the only significant prognostic factors for survival. CONCLUSION: The survival and disease-free survival of patients with diploid DNA content was better compared to aneuploid cases. The results suggest that DNA ploidy status is important in determining the biological behaviour of rectal carcinomas, although the multivariate analysis did not prove its significant influence. The PI were independent negative prognostic factors for survival. PMID- 12552987 TI - A multicenter phase II study of docetaxel and carboplatin combination as front line treatment in advanced non-small cell lung cancer. AB - BACKGROUND: To evaluate the efficacy and safety of docetaxel in combination with carboplatin as first-line treatment of patients with inoperable, locally advanced or metastatic non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Chemotherapy-naive patients with stage IIIB and IV NSCLC, age < 75 years, performance status (WHO) 0-2, were enrolled onto the study. Docetaxel was given at a dose of 100 mg/m2 over an 1-hour i.v. infusion. Carboplatin dosed to an area under the time-concentration curve (AUC) of 6 mg/ml.minute, using the Calvert's formula, was administered over a 30-minute i.v. infusion. The regimen was repeated every 3 weeks. RESULTS: Thirty-eight patients received a total of 155 chemotherapy cycles (median 4 cycles/patient). All patients were assessable for toxicity and 34 for response. There was one (2.6%) complete and nine (23.7%) partial responses; in an intention-to-treat analysis the overall response rate was 26.6% (95% CI: 12.3%-40.3%). The median duration of response was 7 months (range: 3-29), the median time to tumor progression 7 months (range: 3.5-31), and the median overall survival 9 months (range: 0.5-31.5). The probability for 1 year survival was 44%. Grade 3-4 neutropenia was the main hematological toxicity of the regimen occurring in 19 (50%) patients. Four (10.5%) neutropenic episodes were complicated with fever but there was no septic death. Non-hematological toxicity was generally mild. CONCLUSION: These results indicate that the docetaxel-carboplatin combination is a relatively active and well-tolerated front line regimen for the treatment of patients with advanced or metastatic NSCLC. PMID- 12552988 TI - Prognostic factors in Greek patients with small cell lung cancer (SCLC). A Hellenic Cooperative Oncology Group study. AB - BACKGROUND: Prognostic factors among Greek patients with SCLC were evaluated. PATIENTS AND METHODS: Data from 516 patients with SCLC treated by the Hellenic Cooperative Oncology Group were analyzed. Multivariate analysis was performed. RESULTS: Complete response (CR) was achieved in 26.8% and partial response in 40.1% of patients. The median duration of response was 7.4 months, the median survival 10.5 months and the 2-year survival rate 12%. The stage of the disease was a dominant prognostic factor for survival and response. PS 0-1 was a major prognostic factor for survival, duration of response and CR. Female gender was a favorable predictor for CR. Superior vena cava syndrome was a poor prognostic factor for survival and duration of response. Weight loss and age > or = 60 were poor predictors for response. The sites of metastases affected survival, duration of response and response. Normal alkaline phosphatase was a favorable prognostic factor for survival, duration of response and response. Normal lactate dehydrogenase and thoracic irradiation were favorable prognostic factors for survival and duration of response. PMID- 12552989 TI - Prognostic significance of p53, bcl-2 and Ki-67 in high risk superficial bladder cancer. AB - BACKGROUND: Although tumor grade and stage are the most accurate prognostic factors in the evaluation of transitional cell bladder cancer, they cannot always predict the true tumor biological potential since superficial tumors of the same stage and grade may have completely different clinical courses. This study was performed in order to examine whether p53, bcl-2 and Ki-67 have any validity in predicting the course of superficial bladder tumors, with high risk for recurrence or progression, over the traditional prognostic factors that are currently used. Furthermore, we investigated whether any one of these markers maintains its prognostic capability after one course of intravesical instillations of IFN gamma. MATERIALS AND METHODS: The immunohistochemical evaluation of bladder tumor specimens, that were obtained transurethrally for the expression of p53, bcl-2 and Ki-67, was performed in 58 patients. After meticulous selection of cut-off values for the expression of the aforementioned markers, twenty-eight patients were treated only with transurethral resection (TURBT only group) while 30 patients received adjuvantly intravesical instillations of interferon gamma. The times to first recurrence and progression were recorded during the follow-up period which ranged from 3 to 36 months (mean 11.7 months). The prognostic significance of tumor stage, grade, presence of CIS, p53, bcl-2 and Ki-67 in determining the risk for recurrence, was studied with both univariate (log-rank test) and multivariate (Cox regression) methods of analysis, separately for the TURBT only group of patients and for those who received instillations. The same analysis was employed for the risk of progression in the overall number of progressed patients. RESULTS: According to both uni- and multivariate analysis of the prognostic significance for tumor stage (T), grade (G), presence of CIS, p53, Ki-67 and bcl-2 in each group of patients, the Ki-67 index was the only independent prognostic factor for recurrence in patients treated with TURBT only (p = 0.0044 univariate, p = 0.031 multivariate). None of the factors which were studied proved to have prognostic significance for recurrence in the group of patients who received adjuvant intravesical instillation with the immunomodulating agent. Although in the univariate analysis all the studied parameters except tumor stage seem to be associated in a statistically significant manner with higher risk for progression, the multivariate analysis did not yield any independent significant prognosticator. The same evaluation was performed only for the patients with grade 2 disease (28) and yielded statistically significantly higher risk for recurrence, both in uni- (p = 0.0081) and in multivariate analysis (p = 0.044) only in the patients with overexpression of Ki-67 who were treated with TURBT alone. CONCLUSION: The Ki-67 proliferative index has an independent validity in predicting those patients with high risk superficial bladder tumors who may recur in a short follow-up period. A similar relationship of Ki-67 overexpression to progression was not detected. The expression of p53 and bcl-2 does not seem to offer any prognostic information in predicting either recurrence or progression over the prognostic factors that are currently used in clinical practice. PMID- 12552990 TI - Dimethylsilane polyamines: cytostatic compounds with potentials as anticancer drugs. II. Uptake and potential cytotoxic mechanisms. AB - Dimethylsilane tetramines are structural analogues of spermine with a (CH3)2 Si group incorporated into the central carbon chain. They have potential as anticancer drugs. Their cytotoxic effect was considered to rely mainly on their polyamine antagonist property. In order to obtain new ideas about cellular mechanisms, which are potential targets of the dimethylsilane polyamines, the effects of these compounds on some basic cell functions, such as protein and DNA synthesis, and calmodulin antagonism were studied. In addition, their mode of accumulation in cells was investigated. It became evident that the intracellular accumulation of dimethylsilane polyamines is almost exclusively achieved via the polyamine transport system. However, the exchange of a part of the intracellular natural polyamines against dimethylsilane polyamines has only a small effect on polyamine uptake. Binding to the endoplasmic reticulum and inhibition of protein synthesis are presumably important for the cytotoxic action of bis(11-amino-4,8 diazaundecyl)dimethylsilane, a hexamine, but seem of no importance for the tetramines. Calmodulin antagonism, however, is likely to contribute to their cytotoxic effect. PMID- 12552991 TI - Prevalence of breast cancer in women with breast complaints. Retrospective analysis in a population of symptomatic patients. AB - BACKGROUND: The aim of the present study was to analyze whether any correlation exists between breast complaints and the risk of having or developing breast cancer (BC) in a population of self-selected women. PATIENTS AND METHODS: A series of 2,561 patients was reviewed. Three age-groups were sorted out: Group A (40 years old or younger, 45.9%), Group B (41-55 years, 27.8%), and Group C (> 55 years, 26.3%). Breast pain was most common (61.0%) in Group A, and breast lump in Group C (87.8%). RESULTS: BC was found in 271 (10.6%) patients. Breast pain was more frequent in patients without cancer with respect to patients with BC, both in Group B (45.8% vs. 27.9%; p = 0.039) and in Group C (9.2% vs. 1.7%; p = 0.003), but no correlation (p = NS) between BC and breast lump or nipple discharge was found in any Group. Over a median follow-up of 74 months (range 23 146 months). BC arose in 10 (0.44%) patients. CONCLUSION: The majority of the women with breast complaints did not have BC, and their risk of cancer onset was not dissimilar to that reported for the general population. PMID- 12552992 TI - Intrapleural perfusion hyperthermo-chemotherapy under video-assisted thoracoscopic surgery for malignant pleural dissemination: a case report. AB - We report the case of a 45-year-old man treated with intrathoracic hyperthermo chemotherapy (IPHC) under video-assisted thoracoscopic surgery (VATS) for malignant pleural dissemination originating from invasive thymoma. The clinical course of invasive thymoma with malignant pleural dissemination is characterized by early and frequent metastasis and poor survival. In this case, we performed IPHC under VATS with good results. IPHC under VATS is considered a less invasive and more effective approach for malignant pleural dissemination. PMID- 12552993 TI - Radiotherapy with concurrent docetaxel and carboplatin for head and neck cancer. AB - BACKGROUND: This study was designed to evaluate the concomitant use of docetaxel and carboplatin for radiosensitization in head and neck cancer. MATERIALS AND METHODS: One dose of docetaxel at 10 mg/m2 and five doses of carboplatin at AUC of 0.4 per week were administered to patients during the first two weeks of radiotherapy. Sixteen patients were treated with this regimen. Radiotherapy was given to a total dose of 64.8 to 82.0 Gy. Altered fractionation radiotherapy was performed in 12 patients with untreated advanced tumors. RESULTS: The complete response (CR) rate was 81%, with a partial response (PR) rate of 19%. Toxicities included grade 3 mucositis in 69% of patients and grade 2 dermatitis in 56% of patients. CONCLUSION: This schedule of docetaxel and carboplatin combined with radiotherapy may become a useful approach for the management of head and neck cancer with proper management of mucositis. PMID- 12552994 TI - Role of dihydropyrimidine dehydrogenase activity in patients with esophageal cancer. AB - Dihydropyrimidine dehydrogenase (DPD) is the initial and rate-limiting enzyme involved in the degradation of 5-FU. DPD activity in peripheral blood mononuclear cells of 30 esophageal cancer patients treated with 5-FU and low-dose CDDP with irradiation was determined at the beginning of each cytostatic cycle, the objective being to determine if DPD activity is related to the occurrence of side effects and responses to therapy. The DPD activity showed interpatient variability (mean: 325.5 pmol/min/mg protein). 5-FU-related side-effects tended to be registered more frequently in patients with low DPD activity. In particular, nausea occurred in 30.8% of patients in the high DPD activity group but, in 70.6% in those with low DPD activity (p < 0.05). The relationship between the histological response to therapy and DPD activity was nil. We propose that determination of DPD activity prior to initiation of 5-FU-based chemotherapy for patients with esophageal cancer could aid in identifying those at risk for toxicity. PMID- 12552995 TI - Comparison of CYFRA 21-1 and tissue polypeptide specific antigen (TPS) for detecting nasopharyngeal carcinoma. AB - CYFRA 21-1 is a tumor marker that is useful for detecting squamous cell carcinoma (SCC) of the lung. Tissue polypeptide specific antigen (TPS) is a tumor marker that indicates tumor proliferative rate rather than tumor burden. Our aim in this study was to compare the clinical value of CYFRA 21-1 and TPS in the detection of nasopharyngeal carcinoma (NPC). Serum levels of CYFRA 21-1 and TPS were measured in 60 patients with untreated NPC (including 36 differentiated SCC and 24 undifferentiated carcinomas) for comparison with each other. The cut-off values of CYFRA 21-1 and TPS, determined at the 95th percentile of the 43 healthy controls, were 2.50 ng/ml and 115.2 U/l, respectively. The results revealed that the mean serum values of CYFRA 21-1 (6.20 +/- 5.21 ng/ml) and TPS (153.9 +/- 17.3 U/l) in all 60 NPC patients were significantly higher than that in the 43 healthy controls (CYFRA 21-1 = 1.32 + 0.50 ng/ml, TPS = 85.0 +/- 16.0 U/l) (p value < 0.05). In addition, the detecting sensitivities of CYFRA 21-1 (60.0%) and TPS (58.3%) for all 60 NPC were not significantly different (p value > 0.05). In conclusion, our results suggest that both CYFRA 21-1 and TPS are potential tumor markers for the detection of NPC. PMID- 12552996 TI - Risk of primary non-hepatocellular malignancies in hereditary hemochromatosis. AB - OBJECTIVE: The relationship between hemochromatosis, a genetic iron disorder, and the risk of hepatocellular carcinoma is well-documented. However, the true incidence of non-hepatocellular malignancies remains ill defined. We performed a retrospective cohort study to identify a possible association between hereditary hemochromatosis and the development of non-hepatocellular carcinomas. MATERIALS AND METHODS: After appropriate institutional review board approval, an extensive chart review was completed on all patients with hereditary hemochromatosis diagnosed between 1986 and 2001. The diagnosis was based on either a genetic study to identify the Cys282Tyr and the His63Asp mutations on the HLA-H gene and/or hepatic tissue diagnosis, in conjunction with a treatment history of therapeutic phlebotomy. Data collection also included documentation of associated malignancies, the presence of co-morbid medical illnesses, and risk factors for cancer. Our database was age-standardized to the United States population using the National Cancer Institute's Surveillance Epidemiology and End Results (SEER) program data to obtain a standardized incidence ratio (SIR) of non-hepatocellular malignancies in our study population. RESULTS: Fifty-nine patients (49 males, 10 females) with a mean age of 46 years (range: 10-68 years) met the criteria for the diagnosis of hereditary hemochromatosis. Of these, 13 patients (22%) had a biopsy-proven diagnosis of a primary non-hepatocellular malignancy giving an age adjusted SIR of 1.40 (p < 0.04). Histologically, the types of malignancy included: 3 patients with colorectal cancer; 2 patients with gastric carcinoma, 3 patients with prostate cancer; 2 patients with breast cancer; 1 patient with acute myelogenous leukemia, one patient with Hodgkin's lymphoma and one patient with non-Hodgkin's lymphoma. CONCLUSION: While an association between hemochromatosis and hepatocellular carcinoma is well-described, there also appears to be a relationship between hemochromatosis and the development of non hepatocellular malignancies. Since increased iron stores alone have not been proven to be premalignant in recent studies, interplay between genetic factors in hemochromatosis may play a role as a risk factor in the development of an associated malignancy. Larger cohorts of subjects with hereditary iron overload disorders are needed to confirm our findings. Furthermore, if a causal association between hemochromatosis and common carcinomas does indeed exist, such patients may be candidates for individualized, rigorous cancer screening programs. PMID- 12552997 TI - The role of N-acetyltransferase-2 and glutathione S-transferase on the risk and aggressiveness of bladder cancer. AB - N-acetyltransferase-2 (NAT-2) and Glutathione-S-transferase M1 and T1 (GSTM1 and GSTT1) polymorphism have been implicated in the detoxification of urothelial carcinogens, such as arylamines and polycyclic aromatic hydrocarbons. The results of epidemiological studies examining the role of NAT-2, GSTM1 and GSTT1 genotypes on the risk factors for bladder cancer were controversial, although suggesting that there may be an increased risk of the disease associated with these genotypes. The aim of the present study was to examine the independent effect and a possible interaction of NAT-2, GSTM1 and GSTT1 genotypes on the risk of bladder carcinogenesis, in the frame of a case-control study. We also investigated the possible association of specific genotype combinations with more aggressive disease in terms of tumor grading and local staging at the time of initial diagnosis. Between August 1996 and May 1998, 89 newly-diagnosed bladder cancer patients (transitional cell type) and 147 controls were included in the study. All patients were selected at the time of first diagnosis, done in the Department of Urology at the University Hospital of Ioannina, in north-western Greece. GSTM1 and NAT-2 deficient genotypes were found to be independently associated with the risk of bladder cancer (odds ratios 2.87 and 2.64, respectively). The GSTT1 genotype did not present any significant association with bladder cancer risk. We did not find a significant interaction between genotypes. These results could be explained by the independent activity of the two enzymes. Studies that will simultaneously examine the role of several genetic and environmental factors involved in bladder carcinogenesis are needed to give a global picture for the risk factors of bladder cancer and their potential interaction. PMID- 12552998 TI - Lactacystin, a proteasome inhibitor, potentiates the apoptotic effect of parthenolide, an inhibitor of NFkappaB activation, on drug-resistant mouse leukemia L1210 cells. AB - An L1210 cell line (Y8) selected for resistance to deoxyadenosine does not express p53 mRNA or protein but expresses WAF1/p21 even under basal conditions. The Y8 cell line had been previously shown to have an increased apoptotic response to a variety of agents that included DNA damaging agents, kinase inhibitors and drugs directed at NFkappa B activation. In this study we show that lactacystin (LC, an inhibitor of proteasome activity) in combination with parthenolide (PA) caused a synergistic increase in the apoptotic fraction of the Y8 cells. LC (2.5 microM) alone and PA (5.0 microM) caused less than 20% of the Y8 cells to undergo apoptosis. However, the combination of LC (2.5 microM) plus PA (5.0 microM) caused 60% of the Y8 cells to undergo apoptosis. The combination of drugs had no effects on the parental wild-type L1210 cells. Pretreatment of the intact Y8 cells with the caspase-3 inhibitor, Ac-DEVD-CHO, resulted in a marked decrease in the apoptosis caused by the LC plus PA combination. Cell-free extracts prepared from the LC plus PA combination-treated cells had activated caspase activities in the caspase cascade: caspase-3 >> caspase-8 > caspase-6 and caspase-10. These results suggest that there are interacting pathways involving aspects of NFkappa B activation and proteasome activity that could be exploited in therapy directed at p53-deficient tumor cells that would lead to caspase-3 activation and apoptosis bypassing the p53-dependent chemotherapy insensitivity. PMID- 12552999 TI - Different mechanisms of soy isoflavones in cell cycle regulation and inhibition of invasion. AB - BACKGROUND: Soy isoflavones, genistein, daidzein and glycitein, are thought to have beneficial effects on cancer prevention. MATERIALS AND METHODS: We used cell cycle analysis, invasion assay and immunoblotting to determine the effects of genistein and glycitein on Jurkat T cells. RESULTS: Glycitein inhibited Jurkat cell invasion at a level comparable to the inhibition by genistein. Both genistein and glycitein down-regulated MMP-13 proteolytic activity by 60-70% and MMP-8 expression. Caffeine could block G2/M arrest by genistein, but was unable to block the inhibition of invasion by genistein and glycitein. We also demonstrated that glycitein inhibited proteintyrosine phosphorylation in Jurkat cells. CONCLUSION: We determined, for the first time, that glycitein inhibited Jurkat cell invasion, in part through the down-regulation of MMP-13 activity and MMP-8 expression. Our findings also suggest that soy isoflavones may utilize different mechanisms to exert their effects on cell cycle progression and invasiveness of Jurkat cells. PMID- 12553000 TI - p63 expression profile in normal and malignant prostate epithelial cells. AB - p63 expression was evaluated in a tissue microarray of prostate tissues including benign glands, prostatic intraepithelial neoplasia and prostatic carcinoma. Immunostaining with anti-p63 antibody was restricted to basal cell nuclei. Detectable p63 was found in immortalized and early passage cell cultures, but not in senescent cultures or metastatic tumor-derived cell lines. The selective presence of p63 in basal cells of the prostate suggests that p63 can be used as a marker of basal cells and in vitro typing cell cultures of prostate cancer (CaP). The basal cell association underscores its critical functions in the biology of basal cells. PMID- 12553001 TI - DNA topoisomerase II inhibitor, etoposide, induces p21WAF1/CIP1 through down regulation of c-Myc in K562 cells. AB - BACKGROUND: Anticancer agents modulate gene expression and these changes are essential for tumor cell killing. To investigate the mechanism by which etoposide acts as an anticancer agent, the relationship between p21WAF1/CIP1 (p21) and c Myc was studied. MATERIALS AND METHODS: K562 cells with and without ectopic c-Myc expression were studied. Apoptosis was detected using propidium iodide and Hoechst 33342 double staining. The c-Myc and p21 levels were studied by RT-PCR and immunoblot. The p21 promoter (from -205 to +67) was investigated by the luciferase reporter gene assay. RESULTS: Ectopic c-Myc-expressing K562 (K562/c Myc) cells showed more extensive apoptosis than K562 cells after continuous exposure to 200 microM etoposide for 24 hours. During this treatment, p21 expression was not observed in K562/c-Myc cells, and the expression of c-Myc and p21 was mutually exclusive. Etoposide activated the p21 promoter in a concentration-dependent manner, and etoposide-induced luciferase activity was suppressed by co-transfection of c-Myc. CONCLUSION: p21 promoter activity was repressed by c-Myc in proliferating K562 cells, and detoposide-induced down regulation of c-Myc released this suppression, resulting in the induction of p21. PMID- 12553002 TI - Eosinophil-tumor cell interaction in advanced gastric carcinoma: an electron microscopic approach. AB - BACKGROUND: Tumor-associated tissue eosinophils have been observed in human tumors and experimental tumor models, but their function is poorly understood. MATERIALS AND METHODS: One case of intestinal-type adenocarcinoma of the stomach, mainly infiltrated by eosinophils, is studied by light and electron microscopy, focusing on the relationships between eosinophils and tumor cells and on the nature of tumor cell death. RESULTS: Using light microscopy, eosinophils, single or in clusters, were present both in the stroma and within neoplastic glands. With electron microscopy, tumor cells in intimate contact with eosinophils revealed changes consistent with autophagic cell death such as chromatin condensation in small masses into the nucleus, dilation of the nuclear envelope, and numerous cytoplasmic vacuoles. The adenocarcinoma cells, not contacted by neutrophils, remained morphologically well preserved. CONCLUSIONS: Our ultrastructural study suggests the hypothesis of a direct relationship between eosinophil infiltration and induction of autophagic cell death in gastric adenocarcinoma cells. PMID- 12553003 TI - P2P-R deficiency modifies nocodazole-induced mitotic arrest and UV-induced apoptosis. AB - BACKGROUND: Cell cycle progression from G1 through S to mitosis can be influenced by microtubule-dependent mechanisms that involve AP1 factors and c-Jun N-terminal kinase (JNK) activity. UV irradiation-induced apoptosis also involves AP1 factors and JNK activity. The current studies evaluated the outcome of P2P-R deficiency on these mechanisms because P2P-R expression is repressed in association with a decrease in AP1 inducibility and cell cycle progression during differentiation, and P2P-R overexpression promotes apoptosis. MATERIALS AND METHODS: The ability of the microtubule disruption drug nocodazole to induce mitotic arrest and the ability of UV irradiation to induce apoptosis was evaluated in native versus cells made P2P-R deficient by P2P-R antisense treatment. RESULTS: P2P-R deficiency restricts cell cycle progression from G1 through S to mitosis in a microtubule-dependent manner and P2P-R deficiency represses UV irradiation induced apoptosis. CONCLUSION: P2P-R may influence AP1 and/or JNK signaling pathways. PMID- 12553004 TI - Antiproliferative activity of Chinese medicinal herbs on breast cancer cells in vitro. AB - Chinese medicinal herbs are traditionally used to prevent and treat a variety of diseases, including cancer. These herbal preparations are purported to have many biological effects including direct antiproliferative effects on cancer cells, anti-mutagenic activity, and stimulatory or suppressive effects on immune responses. The present study investigates the effects of aqueous extracts from seventy-one Chinese medicinal herbs on the growth of five breast cancer cell lines (SK-BR-3, MCF7, MDA-MB-231, BT-474 and MCNeuA). Twenty-one percent (15 out of 71) of the extracts demonstrated greater than 50% growth inhibition on at least 4 of the 5 cell lines. Dose-response curves were obtained for several of the most potent crude extracts and demonstrated IC50 values ranging from < 10 micrograms/ml to > 1 mg/ml. Six of seven herbs tested induced high molecular weight DNA fragmentation, an early marker of apoptosis, while one of these also induced low molecular weight DNA fragmentation. Flow cytometric analysis of breast cancer cells exposed to one of these herbs (Rheum palmatum) suggested that it arrests cells in the G2/M phase of the cell cycle. These results indicate that many of the herbs used in traditional Chinese medicine for the treatment of cancer have significant growth inhibitory effects on breast cancer cells in vitro. PMID- 12553005 TI - Angiogenesis in benign, pre-malignant and malignant vulvar lesions. AB - Squamous cell carcinoma of the vulva (SCC) accounts for about 4% of all gynaecological malignancies. It has an incidence of about 1.8 per 100,000. However, after the age of 75 the incidence of vulvar carcinoma peaks at approximately 20 per 100,000, making it as common as cervical carcinoma. Benign and pre-malignant vulvar lesions can be broadly divided into non-neoplastic epithelial disorders of the vulva (NNEDV), vulvar intraepithelial neoplasia (VIN) and Paget's disease of the vulva (PDV). NNEDV lesions include lichen sclerosus (LS) and squamous hyperplasia (SH). To date no solid prognostic tools are available to predict which pre-malignant vulvar lesions will progress to SCC. About 4.5% of patients develop SCC following a history of LS and ca. 4% of treated VIN lesions go on to become vulvar SCC. In PDV, 28% of patients have an underlying cancer. Angiogenesis, the development of new blood vessels from existing vasculature, is an essential component of solid tumour growth and metastasis. Several angiogenic factors are expressed by many tumours, suggesting that tumours promote their own vascularisation by activating the host endothelium. This review follows the progress of research in angiogenesis in vulvar disease as assessed by a variety of methods, such as in situ hybridisation (ISH), microvessel density measurements (MVD), immunohistochemically-detected angiogenic growth factor expression, including vascular endothelial growth factor (VEGF) and platelet-derived endothelial cell growth factor/thymidine phosphorylase (PD-ECGF/TP), and serum concentrations of VEGF. Thus, the potential of angiogenesis as a prognostic and predictive tool for identifying which pre malignant lesions could progress to SCC is discussed. A relatively high MVD and strong VEGF expression may serve as prognostic indicators of survival in invasive SCC. MVD and VEGF expression are also predictive factors in identifying which VIN lesions are more likely to become invasive. However VEGF is not a predictive marker in cases of LS likely to progress to carcinoma. The expression of PD ECGF/TP in all types of lesions tested prevents its use as a prognostic tool, unlike VEGF. However, PD-ECGF/TP is involved in PDV pathogenesis, but is not a marker of the malignant progression of PDV. In PDV, VEGF was not expressed even in those cases associated with invasive disease. Serum concentrations of VEGF play a functional role in vulvar carcinogenesis. Although associated with impaired disease-free and overall survival, pre-treatment serum concentrations of VEGF are not an independent predictor of outcome in patients with vulvar cancer. These studies suggest that further angiogenic research will be important in both the therapy and prognosis of malignant and pre-malignant vulvar disease. PMID- 12553006 TI - TGF beta 1 kills lymphoma cells using mitochondrial apoptotic pathway with the help of caspase-8. AB - It is a known paradox that many TGF beta 1-producing tumor cells are resistant to this, otherwise, inhibitory cytokine. In a lymphoma of B-cell origin exogenous TGF beta 1 was able to induce apoptosis, suggesting that the apoptosis program can be switched on. The apoptosis induction was independent of the death receptors but dependent on mitochondrial pathway and caspase-3. Probably due to the weak starting signal, caspase-3 further activated caspase-8 which, through the Bid cleavage and Bax translocation into the mitochondria, provided an autocatalytic support for the apoptotic program. There is a time-gat between the early activation of Smad-dependent TIEG and the accumulation of ROS, therefore other participants that start the increase in mitochondrial membrane permeability should be identified. PMID- 12553007 TI - Molecular analysis of the chemoprotective effects of topical sunscreen and vitamin C in preventing UV-induced and reactive oxygen species-induced DNA damage, respectively, using the PCR inhibition methodology. AB - BACKGROUND: We have developed a PCR-based technique which has previously been shown to be capable of detecting DNA damage (Adducts, strand breaks) induced by carcinogens. We wanted to explore the possibility of detecting anti-carcinogens with this methodology through a corresponding reduction in DNA damage induction. MATERIALS AND METHODS: We have used a PCR-based system which relies on the fact that Taq polymerase cannot amplify damaged DNA efficiently. We immobilised DNA in microtitre plates and exposed them to UV-B light and hydrogen peroxide. In addition, we tested the capacity of sunscreens and ascorbate (vitamin C) to ameliorate the DNA damage caused by UV-B and hydrogen peroxide, respectively. RESULTS: We have shown that in the case of UV-B, sunscreens can effectively reduce the amount of DNA damage induced. This appears to be dependent upon the Sun Protection Factor of the sunscreen, with SPF 35 sunscreens protecting DNA particularly well. We found that both hydrogen peroxide and ascorbate were capable, at high doses, of inducing DNA damage. When mixed, ascorbate effectively increased the DNA damaging effect of hydrogen peroxide. CONCLUSION: We have shown that the PCR inhibition method may be suitable for untangling the complex carcinogenic/anticarcinogenic aspects of factors in the diet and environment. However, the DNA damaging effect of ascorbate must be viewed in the context of the high concentrations used here. PMID- 12553008 TI - Cis 9, trans 11- and trans 10, cis 12-conjugated linoleic acid isomers induce apoptosis in cultured SW480 cells. AB - Dietary conjugated linoleic acid (CLA) has been shown to reduce colon tumor incidence in rodents by mechanisms probably involving apoptosis. The aim of this study was to evaluate the effects of three commercial CLA preparations (pure c9, t11-CLA, pure t10, c12-CLA and a CLA mixture, containing 29.5% c9, t11 and 29% t10, c12-CLA) on caspase-dependent apoptosis in colon SW480 tumor cells. After 4 days incubation, all CLA-treated cells displayed an increase in caspase 3 (27 34%) and caspase 9 activities (37-47%), cleavage of pro-caspase 3 (32 kDa) to 17 and 12 kDa subunits, increased membrane annexin V levels and reduced expression of bcl-2 compared with untreated controls. Cytosolic cytochrome c was increased (p < 0.05) by all CLA preparations, with the t10, c12-CLA isomer being the most potent. The data indicate that t10, c12-CLA may be the more biologically active isomer for inhibition of colon tumor cell proliferation in vitro. PMID- 12553009 TI - Computerized quantitative color analysis for histological study of pulmonary fibrosis. AB - BACKGROUND: Color segmentation is a process through which particular structures can be identified and isolated by the same image according to their color. This may prove particularly useful for the quantification of human pulmonary fibrosis through computerized color segmentation methods applied to trichromic stainings. The aim of this study was to develop and verify a reliable method in order to obtain reproducible parameters of fibrosis using trichromic stainings. MATERIALS AND METHODS: The system used is composed of a PC, hardware and software (Image Pro Plus 4.1-Media Cybernetics, U.S.A.) for image acquisition and analysis, TV camera and optical microscope. The method requires preliminary software procedures of spatial calibration (micron scale) and setting of color segmentation for quantitative color analysis. Massonstained histological specimens of pulmonary parenchyma, with various degree of fibrosis, were analyzed from 10 cases of different lung diseases. RESULTS: Statistical data of the morphological components identified (space, tissue and fibrosis) support the hypothesis that fibrosis distribution depends on relative amount in the tissue and at the same time attest to the precision of the method. CONCLUSION: This procedure could represent a standard method for histopathological evaluation of tissular damage, with fibrosis outcome, especially in connection with the diagnosis and the seriousness of the damage. This would be extremely useful in both the anatomo-pathological and pathological-forensic fields, to make more objective and reproducible quantitative evaluations of pulmonary damage on autopsy or of bioptic material. PMID- 12553010 TI - Adenovirus-mediated anti insulin-like growth factor I gene transfer for the treatment of hepatocellular carcinoma. AB - We have established a hepatocarcinoma cell line (LFCl2A) that produces voluminous tumors when injected into syngeneic Commentary rats. We have previously shown that when these cells were transfected with an episomal vector expressing the antisense IGFI cDNA the transduced cells partly lost their tumorigenic properties and were able to induce the regression of established hepatocarcinoma in syngeneic animals. In this paper, our aim was to determine if one could substitute the use of episomal expression vector by constructing a recombinant adenoviral vector that should be, in theory, easier to supply to humans. We have shown that, in vitro, the cells transfected as well as those infected have lost their tumorigenic properties, but in vivo the infected cells (which are no more tumorigenics) are not able to prevent tumor development. PMID- 12553012 TI - Differential effect of Bcl-xl over-expression on cell death of the monocytic leukemia cell line U937. AB - We studied the effect of over-expression of Bcl-xl on cell death of the monocytic cell line U937. Over-expression of Bcl-xl inhibits apoptotic changes induced by Etoposide including cytochrome-c release, caspase-3 activation and DNA fragmentation. However, Etoposide treatment resulted in cell death in U937 cells over-expressing Bcl-xl, which had a necrotic-like phenotype with no evidence of caspase-3 activation. On the other hand, Bcl-xl over-expression did not prevent U937 cell apoptotic cell death in response to the specific proteasome inhibitor Lactacystin. There was no significant change in the level of Bcl-xl or evidence of its cleavage. These results suggest that Bcl-xl over-expression does not confer protection against cell death in U937 cells and that Lactacystin utilizes an apoptotic pathway not susceptible to Bcl-xl inhibition. PMID- 12553011 TI - The essential 26S proteasome subunit Rpn11 confers multidrug resistance to mammalian cells. AB - BACKGROUND: Identification of multidrug resistance (MDR) factors is crucial for designing chemotherapeutic strategies. Aberrant expression and dysfunction of proteasome subunits have been involved in malignant transformation and in cell resistance to various cytotoxic drugs. MATERIALS AND METHODS: We analyzed the expression levels of the proteasome subunit Rpn11 in a panel of cancer cell lines, and studied the effect of Rpn11 overexpression on the resistance of mammalian cells to cytotoxic drugs in clonogenic cytotoxicity assay. RESULTS: Rpn11 levels are highly variable in cancer cells; mammalian cells stably overexpressing Rpn11 display moderate resistance to vinblastine, cisplatin and doxorubicin, and also exhibit a slower proliferation rate when compared to the control cells. CONCLUSION: Rpn11-overexpression in mammalian cells affects cell proliferation and the response to cytotoxic drugs, both of which may promote tumor cell escape from chemotherapeutic agents, and may serve as a marker for MDR cells. PMID- 12553013 TI - Estrogenic down-regulation of protein tyrosine phosphatase gamma (PTP gamma) in human breast is associated with estrogen receptor alpha. AB - We have reported PTP gamma expression was downregulated by 17 beta-estradiol (E2) and Zeranol (Z) and that PTP gamma may function as an estrogen-regulated cancer suppressor in human breast. We utilized RT-PCR to examine expression of estrogen receptor alpha (ER alpha) and beta (ER beta) mRNA in MCF-7 and MDA-MB-231 cells and to investigate the regulation of PTP gamma expression by E2 and Z in the absence or presence of ICI 182,780 (ICI) in both cells, and immunohistochemistry to examine ER alpha and ER beta protein in normal and cancerous human breast. Results show that MCF-7 express both ER alpha and ER beta, and MDA-MB-231 express only ER beta. Both E2 and Z (30 nM; 24 h) suppressed PTP gamma by approximately 56% in MCF-7 cells and these effects were completely blocked by 1 mM of ICI. In contrast, E2, Z and ICI had no effects on PTP gamma expression in MDA-MB-231 cells. Interestingly, both E2 and Z suppressed PTP gamma by approximately 45% in MDA-MB-231 cells transfected with ER alpha, and these effects were completely blocked by 100 nM of ICI. Both RT-PCR and immunohistochemical staining showed that ER alpha expression was significantly higher in cancerous human breast than in normal breast, while ER beta was higher in normal human breast than in cancerous breast. In combination with our previous findings of greater PTP gamma expression levels in normal human breast than cancerous breast, current results show that lower PTP gamma was associated with higher ER alpha in cancerous human breast tissues. In conclusion, results indicate that Z induces estrogenic effects in human breast relative of PTP gamma expression and the estrogenic down regulation of PTP gamma expression in human breast is associated with ER alpha. PMID- 12553014 TI - A new phenylacetate-bisphosphonate inhibits breast cancer cell growth by proapoptotic and antiangiogenic effects. AB - Sodium phenylacetate (NaPa) and some bisphosphonates demonstrated antiproliferative and proapoptotic properties against cancer. We have previously shown that NaPa inhibited cell proliferation of MCF7-ras tumor breast cells both in vitro and in vivo. On the other hand, bisphosphonate activities have only been demonstrated in vitro. Here we evaluated the antitumor effects of a new bisphosphonate, the phenylacetate-bisphosphonate (PaBp), on human breast cancer MCF7 and MCF7-ras cell lines, both in vitro and in vivo. To our knowledge, this is the first report indicating the use of a bisphosphonate derivative as a powerful cytostatic and cytotoxic agent, with proapoptotic and antiangiogenic properties on human breast cancer cells lines, with no animal toxicity. PMID- 12553015 TI - Anti-tumor and anti-vascular effects of the novel tubulin-binding agent combretastatin A-1 phosphate. AB - The combretastatins are derived from an African medicinal plant Combretum caffrum (Combretaceae). They have previously been shown to be potent inhibitors of microtubule assembly that cause marked haemorrhagic necrosis in murine subcutaneous tumors. Promising clinical trial results with combretastatin A-4 phosphate led to this investigation of the anti-tumor and anti-vascular effects of a close structural analog, combretastatin A-1 phosphate. This compound caused identical disruption of the tubulin cytoskeleton in HUVECs in vitro at similar concentrations and duration of exposure as combretastatin A-4 phosphate. Treatment of a well-vascularised murine colon adenocarcinoma (MAC 29) with an effective dose (150 mg/kg) of combretastatin A-1 phosphate resulted in a dramatic decrease in functional vascular volume 2 hours after administration. Vascular shutdown was complete within 4 hours after treatment apart from in small areas of the tumor periphery. Morphological examination of hepatic deposits of HT29 and DLD-1 human colon tumors in nude mice demonstrated that combretastatin A-1 phosphate displays greater anti-tumor effects than the A-4 analog at the same dose and this order of activity (A-1 > A-4) is mirrored in the subcutaneous site with the same tumor type. In summary, combretastatin A-1 phosphate can exert its anti-tumor action via an anti-vascular mechanism. The results indicate that, despite having similar in vitro anti-tubulin properties, combretastatin A-1 phosphate seems to have greater in vivo anti-tumor activity than combretastatin A 4 phosphate at the same doses and may therefore be more successful in the clinic. PMID- 12553016 TI - Expression and role of thrombospondin-1 in colorectal cancer. AB - Thrombospondin-1 (TSP1) inhibits angiogenesis and activates transforming growth factor beta-1 (TGF beta-1). The expression and role of TSP1 remain controversial. On 132 colorectal cancer specimens, we performed immunohistochemical staining of TSP1, TGF beta-1, latency-associated peptide (LAP) and CD34, besides performing in situ hybridization (ISH) of TSP1. TSP1 was mainly localised in fibroblasts of the tumor stroma on ISH. The result revealed that 73 cases (55.3%) were evaluated as high-TSP1 while 84 cases (63.6%) were evaluated as high-TGF beta-1. The expression of TSP1 correlated significantly with vessel counts (p = 0.016) and TGF beta-1 expression (p = 0.043). Cox proportional hazard analysis showed that TSP1 expression was significantly correlated with independent prognostic factors. The present study furnishes evidence indicating that TSP1 is expressed in tumor stroma, inhibits tumor angiogenesis and suppresses tumor growth by activating TGF beta-1. PMID- 12553017 TI - Does a low frequency of P53 and Pgp expression in familial glioma compared to sporadic controls indicate biological differences? AB - BACKGROUND: A three-fold increased risk for glioma among first degree relatives (FDR) to glioma patients has previously been shown. This study compared familial cases with sporadic controls of glioma to see if phenotypic differences could be detected. Different pathways to tumour growth and progression were investigated including cell cycle regulating genes (p53) and growth factors (epidermal growth factor receptor, EGFR), angiogenesis (vascular endothelial growth factor, VEGF and microvessel density, MVD), pathways of detoxification (glutathione-S transferase, GST pi) and multidrug resistance (P-glycoprotein, Pgp). MATERIALS AND METHODS: Thirty-seven cases of familial gliomas, identified in a population based study, were compared to 58 sporadic glioma controls chosen with a negative family history of glioma. The immunohistochemistry was performed with standard procedures using the LABSA kit (Zymed lab). RESULTS: Familial cases had significantly more frequent p53- and Pgp-negative tumours, also when correcting for age, sex and histopathology. However, Pgp was no longer significant after correcting for p53 status indicating a correlation between Pgp and p53. A significant difference between VEGF-negative to VEGF-positive tumours (low- or high-grade) was shown, but it was no longer significant when correcting for p53 status. CONCLUSION: Our study investigated phenotypic differences of familial glioma compared to sporadic control. Our finding of a distinct pattern of increased p53- and Pgp-negativity among cases warrants further investigation. PMID- 12553018 TI - The more an ulcerative colitis is repeated, the more the risk of colorectal carcinogenesis is increased in mice. AB - The incidence of colorectal carcinogenesis is increased in patients with ulcerative colitis. We investigated the effects of repetitive diarrhea on colorectal carcinogenesis in mice singly pretreated by a low-dose of chemical carcinogen. Mucosal changes were investigated in mice pretreated with a single intraperitoneal injection of 8.0 mg/kg body weight of azoxymethane prior to a repetitive oral administration of 3% dextran sulphate sodium to induce chronic diarrhea. Repetitive treatment with dextran sulphate sodium induced a cycle of chronic diarrhea-remission in the colorectum. Five submucosal-invasive adenocarcinomas and 65 high-grade dysplasias were found in 15 mice that underwent azoxymethane-pretreatment with 3 cycles of 3% dextran sulphate sodium-exposure, i.e. colorectal carcinogenesis was observed mainly in the left side of the large intestine within 11 weeks after the initial treatment with carcinogen. Consequently, thymidine kinase was expressed and activated, and an increase in bromodeoxyuridine-immuno-reactive (S-phase) cells was observed in the regenerating mucosa and the colorectum with tumorous lesions. Colorectal carcinogenesis developed with the increasing duration of diarrhea induced by 3% dextran sulphate sodium in mice pretreated by a single injection of a small dosage of azoxymethane during the comparatively short period of this experimental colitis system. PMID- 12553019 TI - Overexpression of early growth response-1 as a metastasis-regulatory factor in gastric cancer. AB - BACKGROUND: To investigate the potential role of a nuclear transcription factor, early growth response-1 (Egr-1), in formation and progression of gastric cancer, we compared its expression in gastric cancers with that in non-cancerous tissues. MATERIALS AND METHODS: Egr-1 mRNA expression was measured using TaqMan RT-PCR. The corresponding protein expression was examined immunohistochemically. RESULTS: Egr-1 mRNA expression was significantly higher in gastric cancer tissues than in normal mucosa (p < 0.0005). These differences were also reflected by protein product expression. Moreover, Egr-1 mRNA expression was higher in cases with metastasis to lymph nodes or remote organs. In cultured gastric cancer cells known to have a high metastatic potential, expression of this mRNA was higher than that of parental cells. CONCLUSION: It was suggested that Egr-1 has a significant role in carcinogenesis and in cancer progression, especially metastasis. Measurement of this mRNA should be useful for evaluation of the metastatic potential of gastric cancer. PMID- 12553020 TI - The correlation between CpG methylation on promoter and protein expression of E cadherin in oral squamous cell carcinoma. AB - BACKGROUND: Reduction of E-cadherin in most common epithelial tumors relates to metastasis, which results from the silence of E-cadherin by CpG methylation. MATERIALS AND METHODS: We examined the E-cadherin expression by immunohistochemical staining and detected methylation by methylation-specific polymerase chain reaction (MSP) in 48 primary oral SCC tissues. RESULTS: The results showed that 41 out of 48 (85.4%) cancerous tissues and 16 out of 48 (33.3%) nearby non-cancerous tissues had CpG methylation on the promoter region of E-cadherin. In these non-cancerous tissues, 2 out of 16 (12.5%) had no methylation change in their paired cancerous part. Immunohistochemical study showed that a decreased expression pattern was found in the tissue which had CpG methylation on the promoter region, but an over expression island or aberrant expression was also frequently found in these cases. CONCLUSION: The methylation of E-cadherin in oral SCC may occur in the precancerous stage and the process is dynamic, which has no relationship with the aberrant expression of E-cadherin protein. PMID- 12553021 TI - Expression of vascular endothelial growth factor (VEGF) and its receptors (Flt-1 and Flk-1) in esophageal squamous cell carcinoma. AB - BACKGROUND: To clarify the clinical significance of vascular endothelial growth factor (VEGF) and its receptors, VEGFR-1 (Flt-1) and VEGFR-2 (Flk-1), in esophageal cancers, we evaluated the relationships between the expression of VEGF and its receptors, tumor microvessel density (MVD), clinicopathological factors and prognosis. MATERIALS AND METHODS: Thoracic esophageal squamous cell carcinoma (SCC) specimens were obtained during surgery from 64 previously untreated patients. Expression levels of VEGF and its receptors were determined immunohistochemically, and tumor MVD was assessed. RESULTS: Twenty-four (37.5%) of the tumors showed diffuse VEGF immunoreactivity. VEGF expression was significantly correlated with tumor status (p < 0.05) and poor prognosis (log rank; p < 0.05). VEGFR-1 immunoreactivity was detected in the cytoplasm of the cancer cells in 27 patients (42.2%). VEGFR-1-positive cancers tended to be associated with poorer nodal status (p = 0.1513), but VEGFR-1 expression did not correlate with prognosis in the univariate survival analysis (p = 0.2964). VEGFR 2 immunoreactivity was detected in the cancer cell cytoplasm in 26 patients (40.6%), but VEGFR-2 expression did not correlate with clinicopathological factors or prognosis. Comparison of the MVD and clinicopathological characteristics revealed significant associations between high MVD and poorer tumor status (p < 0.05), blood vessel invasion (p < 0.05) and poor prognosis (p < 0.01). In the multivariate analysis, MVD was identified as an independent prognostic factor as well as depth of tumor invasion and lymph node metastasis. Although VEGF expression correlated significantly with the MVD (p < 0.05), the correlations between VEGFR-1 and VEGFR-2 expression and the MVD were not significant. There were, however, significant intercorrelations in expression between VEGF, VEGFR-1 and VEGFR-2. CONCLUSION: Vascular endothelial growth factor (VEGF) may play a role in the control of angiogenesis in esophageal squamous cell carcinoma. However, although VEGF expression correlates significantly with coexpression of its receptors, VEGFR-1 and VEGFR-2, these receptors do not appear to contribute directly to tumor progression. Nevertheless, VEGF and its receptors represent a logical target for antiangiogenic therapy for esophageal SCC. PMID- 12553022 TI - A matrix metalloproteinase inhibitor, ONO-4817, retards the development of mammary tumor and the progression of uterine adenomyosis in mice. AB - The inhibitory effects of a novel matrix metalloproteinase inhibitor, ONO-4817, on the development of mammary tumors and the progression of uterine adenomyosis were examined in SHN mice. First, multiparous mice which developed mammary tumors spontaneously were used. The first palpable tumor was removed, and the mice were thereafter fed chow containing ONO-4817. Any second mammary tumor developing in the other mammary fat pad was also removed, and the mice were continuously fed the chow containing ONO-4817. The mice were killed when a third tumor was detected in the other fat pad. The periods between the occurrence of the first and second tumors, and the second and third ones were significantly increased in the mice treated with ONO-4817 compared to the mice not given ONO-4817 treatment. Second, to test ONO-4817 suppression of the progression of the invasion of uterine adenomyotic tissue, mice with experimentally-induced adenomyosis were treated with ONO-4817 for 4 weeks. The degree of pathological progression of adenomyosis was less in the uteri exposed to ONO-4817 than in the uteri not exposed to the inhibitor. PMID- 12553023 TI - Boron-lipiodol: a potential new drug for the treatment of liver tumors. AB - BACKGROUND: Boron neutron capture therapy (BNCT) is a form of radiation therapy and has been proposed for the treatment of some malignancies with encouraging results. However, none of them has ever been applied to liver malignancy. The purpose of this study was to evaluate the potential of boron-lipiodol (B lipiodol) for the treatment of VX2 liver tumor via BNCT. MATERIALS AND METHODS: Twelve New Zealand rabbits were randomly separated into two groups: lipiodol and boron-lipiodol groups. The rabbits were anesthetized, a midline incision was made and the left lobe of the liver was injected with 0.1 ml of VX2 tumor cells. After the tumor reached 2-3 cm in diameter, the rabbits were anesthetized and 0.5 ml of boron-lipiodol was injected into the hepatic artery via an angiocatheter. Liver function tests and renal function tests were performed before, at 12 hours, 24 hours, 48 hours and 7 days after injection of drugs in both groups. The concentration of boron in various tissues was determined on the 7th day after injection. RESULTS: Liver function was abnormal at 12 hours after injection, and then gradually returned to normal at 7 days, indicative of acute temporary hepatic damage. As for the renal function, no significant change was noted in either group. The boron level was 49.7 ppm in tumor and 6.31 ppm in the healthy liver 7 days after injection of B-lipiodol. The ratio of boron concentrations between the tumor and the normal liver tissue was 7.87. As for blood and other organs including spleen, heart and kidney, the concentration of boron was low. In the lipiodol group, the boron concentrations in tumor and various organs were low. CONCLUSION: The high concentration of boron after intra-arterial injection of B-lipiodol can be used for neutron capture therapy. B-lipiodol has potential for the treatment of liver malignancy. PMID- 12553024 TI - Inhibition of endometrial carcinoma cell growth using antisense estrogen receptor oligodeoxyribonucleotides. AB - BACKGROUND: The sex hormone 17 beta-estradiol acts as a mitogen in a number of tissues, including the endometrium, through direct interaction with the estrogen receptor (ER alpha). In the protection of the female breast and endometrium from cancer progression it would be advantageous to inhibit estrogenic action, therefore many estrogen receptor antagonists have been made. However, the most clinically relevant anti-estrogens for breast cancer have a detrimental effect on the endometrium and induce or exacerbate existing endometrial oncogenesis. Specific anti-estrogenic potential that is directed against the endometrial ER alpha theoretically could be achieved using antisense oligodeoxyribonucleotide transfection techniques. MATERIALS AND METHODS: To discover the most effective antisense oligodeoxyribonucleotides against the human ER alpha, a series of oligodeoxyribonucleotides were synthesised and tested in a human endometrial cancer cell line that expresses a functional ER alpha. RESULTS: Transfection with antisense oligodeoxyribonucleotides, directed against different regions of the human ER alpha, significantly inhibited ER alpha protein production without affecting ER beta protein levels. Antisense oligodeoxyribonucleotides directed against the translational start site demonstrated reduced binding of radiolabeled 17 beta-estradiol and a complete inhibition of estrogen-dependent endometrial cancer cell proliferation. The inhibitory effect of the antisense oligodeoxyribonucleotides on ER alpha production and ligand binding was enhanced in the presence of exogenous 17 beta-estradiol. CONCLUSION: The data suggest that antisense oligodeoxyribonucleotides against the human ER alpha have the potential of acting as anti-proliferative agents for estrogen-dependent endometrial cancers and may help in elucidating the relative roles of the two estrogen receptors, ER alpha and ER beta, in cell processes other than proliferation. PMID- 12553025 TI - Chemoprevention of oral cancer by black raspberries. AB - Oral cavity cancers represent 2.5% of the cancers that occur in the United States and are ranked sixth worldwide. Since current therapeutic protocols are relatively ineffective, alternative strategies for prevention need to be developed and tested in appropriate animal models. In the study reported herein, the hamster cheek pouch (HCP) was used to evaluate the ability of black raspberries to inhibit oral cavity tumors. Male Syrian Golden hamsters, 3-4 weeks of age, were fed 5% and 10% lyophilized black raspberries (LBR) in the diet for two weeks prior to treatment with 0.2% 7,12-dimethylbenz(a) anthracene in dimethylsulfoxide and for 10 weeks thereafter. HCPs were painted 3X/week for eight weeks. The animals were sacrificed 12-13 weeks from the beginning of DMBA treatment and the number and volume of tumors (mm3) determined. There was a significant difference (p = 0.02) in the number of tumors between the 5% LBR and control groups (27 tumors/14 animals and 48 tumors/15 animals, respectively) and an intermediate number of tumors in the 10% berry-treated animals (39 tumors/15 animals). These experiments support previous studies from our laboratories showing the chemopreventive activity of black raspberries and show, for the first time, that dietary black raspberries will inhibit tumor formation in the oral cavity. PMID- 12553027 TI - Comparative antitumor activity of 5-fluorouracil and 5'-deoxy-5-fluorouridine in combination with interferon gamma in renal cell carcinoma cell lines. AB - The treatment of advanced renal cell carcinoma (RCC) remains unsatisfactory, and conventional therapy using interferon (IFN) has shown limited antitumor action. In this study, we examined the combined effects of IFN gamma with 5-fluorouracil (5-FU) as well as 5'-deoxy-5-fluorouridine (5'-DFUR), a prodrug of 5-FU, in vitro. The sensitivity of 4 RCC cell lines to IFN gamma and 5-FU or 5'-DFUR was evaluated using the MTT assay. The IC50 value for the cytostatics was expressed as the concentration at which growth was inhibited by 50% as compared with the control value. Thymidine phosphorylase (TP), the enzyme that converts 5'-DFUR into 5-FU and 5-FU into 5-fluoro-2'-deoxyuridine-5'-monophosphate (FdUMP), was estimated by ELISA. As a result, a significant correlation between TP expression and sensitivity to 5-FU and 5'-DFUR was found in all 4 RCC cell lines. By adding 12.5 U/ml of IFN gamma, which is a concentration that does not affect cell proliferation, the TP expression significantly increased by 1.5 to 3.4 times in the 4 RCC cell lines. Susceptibility to 5-FU and 5'-DFUR was also significantly induced by 1.42 to 2.36 times and 2.9 to 5.71 times, respectively, in the 4 cell lines. These results suggested that TP may be useful as a predictive factor in combination therapy with IFN gamma and 5-FU or 5'-DFUF, which may be a promising treatment for advanced RCC. PMID- 12553026 TI - Interaction between dental metals and antioxidants, assessed by cytotoxicity assay and ESR spectroscopy. AB - Among dental metals, copper showed the highest cytotoxicity against human oral squamous cell carcinoma and human submandibular gland carcinoma cells, followed by palladium-alloy, gold and silver. Normal human cells (gingival fibroblast, pulp cells, periodontal ligament fibroblast) were relatively resistant to these metals. The palladium-alloy failed to induce internucleosomal DNA fragmentation, a biochemical hallmark of apoptosis, in human promyelocytic leukemic HL-60 cells. The cytotoxic activity of the palladium-alloy was significantly reduced by a non cytotoxic concentration of N-acetyl-L-cysteine, or more efficiently by sodium ascorbate. However, higher concentrations of sodium ascorbate enhanced the cytotoxic activity of palladium-alloy. ESR spectroscopy showed that the palladium alloy enhanced the intensity of ascorbate radical, suggesting the possible interaction between metals and antioxidants. All metals, except copper, did not significantly affect the generation of superoxide anion (by hypoxanthine-xanthine oxidase reaction), hydroxyl radical (by Fenton reaction) and nitric oxide (from NOC-7 in the presence of C-PTIO). These data demonstrate for the first time that antioxidants modify the biological activity of dental metals. PMID- 12553028 TI - Topoisomerase inhibitor-induced apoptosis accompanied by down-regulation of Bcl-2 in human lung cancer cells. AB - BACKGROUND: Anticancer drug-induced apoptosis is one of the important mechanisms in the effectiveness of chemotherapy. In this study, we investigated apoptosis induced by topoisomerase inhibitors and its relationship with chemosensitivity, the expression of Bcl-2 family proteins and p53 status in human lung cancer cells. MATERIALS AND METHODS: A total of six human lung cancer cell lines, i.e., two small cell lung cancer (SCLC) and four non-small cell lung cancer (NSCLC) cell lines, were used. For induction of apoptosis, these cell lines were treated with SN-38 (DNA topoisomerase I inhibitor) or etoposide (DNA topoisomerase II inhibitor). Drug sensitivity was determined using a microculture tetrazolium assay. The rates of apoptosis and alterations of Bcl-2 and Bax expression were analyzed by flow cytometry. RESULTS: Apoptotic cells increased in a time dependent manner after exposure to topoisomerase inhibitors. Induction of apoptosis was accompanied by the down-regulation of Bcl-2 expression, but there was little alteration of Bax expression. These events were significantly more extensive in SCLC cell lines, which are more sensitive to topoisomerase inhibitors, than in NSCLC cell lines, which are more resistant to these inhibitors. However, neither induction of apoptosis nor chemosensitivity correlated with p53 status in the lung cancer cell lines studied. CONCLUSION: The more extensive induction of apoptosis with Bcl-2 down-regulation in SCLC than in NSCLC might explain, at least in part, the higher clinical sensitivity to topoisomerase inhibitors in the former disease. PMID- 12553029 TI - Expression of surface protein receptors in lung cancer. AB - Cell adhesion is a basic count in inter- and intracellular communication and plays an important role in tumor progression. In this study, the expression of E selectin, ICAM-1 and VCAM-1 were evaluated by means of immunohistochemistry in a group of 153 lung cancer specimens. E-selectin immunoreactivity was localized mostly on endothelial cell venules and capillaries with an average staining intensity of 75% of cells in the NSCLC, while in SCLC the intensity of the staining was 69%. The staining pattern for ICAM-1 reached an average intensity of 57%, in both NSCLC and SCLC. Finally, VCAM-1 immunoreactivity was detected only in NSCLC with an average intensity of 12% on endothelial cell venules and capillaries. This study provides a contribution towards the understanding of the basic mechanisms of cell adhesion in lung cancer progression. PMID- 12553030 TI - Vitamin D3 up-regulated protein 1 (VDUP1) expression in gastrointestinal cancer and its relation to stage of disease. AB - Decreased expression of VDUP1, which is an interesting stress response gene, has been shown in rat mammary tumors and has been discussed in relation to the development of the tumor. However, VDUP1 expression in clinical specimens of human cancer remains unclear. We employed TaqMan RT-PCR assay to investigate VDUP1 expression in surgical specimens of primary tumors and their adjacent normal tissues from gastrointestinal cancer patients, 40 with colorectal and 12 with gastric cancers. TaqMan RT-PCR assay showed that VDUP1 expression in colorectal and gastric cancers was significantly lower than that in their adjacent normal tissues (p < 0.0001 and p < 0.001, respectively). In addition, we found that VDUP1 expression was associated with clinical stage in colorectal cancer (p < 0.01). VDUP1 expression in stage II patients was significantly higher than that in stage III (p < 0.05) and in stage IV patients (p < 0.01). These results suggest a possible role of VDUP1 in the pathogenesis of gastrointestinal cancer, as well as its clinical significance. PMID- 12553031 TI - Cationic liposomes with plasmid DNA influence cancer metastatic capability. AB - BACKGROUND: Cancer gene therapy is now being developing to provide new strategies for the treatment of human tumors. Cationic lipids represent one of the powerful mediators for DNA delivery. The liposome-plasmid DNA complex itself is known to inhibit tumor cell growth, but other effects on cancer cell behaviors have not been reported so far. MATERIALS AND METHODS: Six commercially available cationic liposomes complexed with plasmid DNA were applied to cancer cells and their metastatic potentials were measured. RESULTS: The liposome-plasmid DNA complexes affected metastatic capability in three different ways: TM-TPS:DOPE and DOTAP:DOPE had no effect on metastatic capability; a suppressive effect was observed in DOSPA:DOPE and DMRIE:cholesterol; while an augmentative effect was observed in DOTMA:DOPE and Effectene. These effects are likely to be DNA sequence independent, because different plasmids have the same effects. CONCLUSION: Liposome-plasmid DNA complexes influence cancer metastasis capability, dependent upon the cationic liposome formulations. PMID- 12553032 TI - Inhibitory effect of glycyrrhizin on experimental pulmonary metastasis in mice inoculated with B16 melanoma. AB - The pulmonary metastasis of B16 melanoma cells has been shown to be influenced by tumor-associated T helper type 2 cells (Th2 cells). Glycyrrhizin (GR), an active component of licorice roots, has been described as an inducer of CD4+ CD28+ TCR alpha/beta+ T cells, which are capable of counteracting Th2 cytokine production by Th2 cells. In the present study, the anti-metastatic effect of GR in mice implanted with highly metastatic B16F10 melanoma cells was investigated. When mice inoculated with B16F10 melanoma cells (1 x 10(5) cells/mouse, i.v.) were treated with GR (10 mg/kg) 1, 3, 5 and 7 days after the tumor inoculation, the incidence of metastases decreased on average; 48 colonies were found in the GR treated group and 208 in the saline-treated group. Similarly, 84% of metastases were inhibited in these mice after they were inoculated with splenic CD4+ T cells from mice treated with GR (GR-CD4+ T cells). These results suggest that GR inhibits the pulmonary metastases of B16 melanoma through the regulation of tumor associated Th2 cells. PMID- 12553033 TI - Endogenous tumor necrosis factor promotes resistance to cellular stresses by inducing the metallothionein-1A gene. AB - BACKGROUND: Endogenous tumor necrosis factor (enTNF) acts as a resistance factor against anticancer drugs, heat and exogenous TNF via induction of manganous superoxide dismutase (MnSOD) and heat shock protein 72 (HSP72), while the details of interaction with other molecules are not fully understood. We compared mRNA expression of various genes between MIAPaCa-2 human pancreatic cancer cells and M5 cells transduced with nonsecretory-type TNF gene expression vector. MATERIALS AND METHODS: The expression of certain mRNAs between human pancreatic cancer MIAPaCa-2 cells and M5 cells was compared using fluorescent differential display. RESULTS: Of 140 bands obtained by gel electrophoresis, 53 bands showed patterns differing between MIAPaCa-2 and M5 cells. Among these bands, sequence analysis and RT-PCR identified strong mRNA expression for metallothionein-1A, a scavenger of reactive oxygen species, in transduced M5 cells. CONCLUSION: Metallothionein 1A could be induced by enTNF, resulting in resistance against various cellular stresses. PMID- 12553034 TI - Induction of Epstein-Barr virus (EBV) reactivation in Raji cells by doxorubicin and cisplatin. AB - BACKGROUND: Epstein-Barr virus (EBV) can be activated in B-lymphoid cells to enter the lytic cycle by various kinds of stimuli, including 12-O tetradecanoylphorbol-1 3-acetate (TPA), butyric acid, calcium ionophore A23187, transforming growth factor-beta and anti-immunoglobulin crosslinking. EBV reactivation has been clinically observed in patients receiving systemic chemotherapy. This study sought in vitro evidence to suggest whether anticancer drugs may directly contribute to the EBV reactivation. MATERIALS AND METHODS: Raji cells, an EBV-containing Burkitt's lymphoma cell line, were used as the experimental model. TPA served as a positive control for chemical induction of EBV reactivation. Expression of the BZLF1 transcript of EBV and its encoded protein, ZEBRA, were examined by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and flow cytometry, respectively. Transactivation activity of ZEBRA was further assessed by a luciferase reporter assay of EBV DR-promoter activity and a flow cytometry assay assessing the endogenous expression of EA-D (BMRFl). RESULTS: Doxorubicin and cisplatin, two commonly used anticancer agents, induced a dose-dependent up-regulation of BZLF1 mRNA and ZEBRA protein. The luciferase reporter activity and the expression of endogenous EA-D protein, also increased by doxorubicin and cisplatin, indicated an up-regulation of the transactivating activity of ZEBRA. CONCLUSION: These data indicate that cytotoxic anticancer drugs may up-regulate the expression and the transactivating activity of BZLF1, and suggest that systemic chemotherapy may be a risk factor for EBV reactivation in patients with EBV-associated malignancies. PMID- 12553035 TI - Ex vivo chemosensitivity to mitomycin C in bladder cancer and its relationship with P-glycoprotein and apoptotic factors. AB - BACKGROUND: Intravesical treatment with mitomycin C (MMC) leads to a complete response rate of around 40% in superficial bladder cancer (TCC). In order to determine in advance which patients will fail to respond, we describe a study assessing the feasibility of applying the ATP assay to test the chemosensitivity of samples from patients with this disease. MATERIALS AND METHODS: TURBT or biopsy samples were received from 27 patients, 23 of which were suitable for the ATP assay (16 primary tumours and 7 recurrences). RESULTS: The success rate of the assay was 91%. There was a marked variation in the effect of MMC between patients with a > 50-fold range in LC50 values (drug concentration required to kill 50% of cells) from 2.99- > 150 microM with a median value of 22.4 microM. We were unable to determine any overall correlation between chemosensitivity and tumour stage or grade or the treatment status of the patient in this small data set. P-glycoprotein status and caspase-3 levels were assessed on these samples using immunohistochemistry but there did not appear to be any relationship between either of these parameters and MMC resistance. Apoptotic counts and mitotic counts were also measured but, whilst these appeared to correlate with grade (p < 0.01), there was no overall significant relationship established with MMC chemosensitivity. CONCLUSION: This study suggests that it is possible to use the ATP assay for chemosensitivity testing in TCCs. Despite a lack of overall correlation between ex vivo MMC resistance and the conventional prognostic factors tested, further studies are warranted in a larger data set to test the ability of this technique to predict clinical outcome in this disease. PMID- 12553036 TI - Nuclear-factor kappa B (NF-kappa B)-inducible nitric oxide synthase (iNOS/NOS II) pathway damages the stria vascularis in cisplatin-treated mice. AB - BACKGROUND: Cisplatin is reported to damage the stria vascularis of the cochlea. Free radicals, especially large amounts of nitric oxide catalyzed by inducible nitric oxide synthase, are considered to have an important role in this toxicity. The induction of inducible nitric oxide synthase is regulated by nuclear-factor kappa B (NF-kappa B). We examined the damage of the stria vascularis by immunohistochemical techniques. MATERIALS AND METHODS: Cisplatin (15 mg/kg b.w.) was injected intraperitoneally into the mice. Three days after the injection, the cochleas were immunohistochemically-stained using specific antibodies for nuclear factor kappa B (NF-kappa B), inducible nitric oxide synthase (iNOS) or single stranded DNA. RESULTS: NF-kappa B was expressed in the cisplatin-treated cochlea, especially in the stria vascularis and the spiral ligament. iNOS was also expressed in the stria vascularis and the spiral ligament. Fragments of DNA were observed only in the stria vascularis. CONCLUSION: The large amounts of NO catalyzed by iNOS led to inner ear dysfunction. Our results indicate that apoptosis is triggered by iNOS and that it mediates the ototoxicity induced by cisplatin. PMID- 12553037 TI - Enhanced expression of the UROC28 gene in human breast cancer: relationship to ERBB2 gene expression. AB - BACKGROUND: ERBB2, a highly important oncogene in invasive breast cancer, is not only a prognostic factor but also a predictive marker for response to therapeutic agents. Recently, An et al. (10) identified a novel gene, UROC28, that is also overexpressed in breast cancer. To examine possible interrelationships, we quantitated UROC28 and ERBB2 mRNA in breast cancers. MATERIALS AND METHODS: The expression of UROC28 and ERBB2 mRNA in breast cancer tissues were determined using RT-PCR. The expression was also examined in T-47D breast cancer cells treated with estrogen. RESULTS: UROC28 mRNA expression was greater in cancers than in noncancerous tissues (p < 0.0001), as was ERBB2 mRNA. ERBB2 and UROC28 gene expression was dose-dependently down-regulated in T-47D breast cancer cells treated with estradiol. However UROC28 mRNA and ERBB2 mRNA expression did not correlate with one another. CONCLUSION: These results indicate that UROC28 may be a useful target molecule in breast cancer diagnosis and treatment, complementing ERBB2. PMID- 12553038 TI - Expression of IL-6 receptor in pancreatic cancer: involvement in VEGF induction. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is one of the most potent angiogenic growth factors and its expression is correlated with MVD (microvascular density) in malignant tumors, including pancreatic adenocarcinoma. On the other hand, serum interleukin-6 (IL-6) is elevated in many patients with pancreatic cancer in accordance with their disease progression. In this study, we examined whether IL-6 and its receptors have any involvement in the induction of VEGF in pancreatic cancer. MATERIALS AND METHODS: Two human pancreatic cancer cell lines were examined for the induction of VEGF after treatment with IL-6. Thirty-two pancreatic cancer surgical specimens were stained immunohistochemically with VEGF, IL-6 and IL-6 receptor antibodies. RESULTS: CFPAC-1 cells expressed IL-6 receptor whereas AsPC-1 cells rarely expressed it. IL-6 treatment induced VEGF expression significantly and dose-dependently in CFPAC-1 cells, while it did not change in AsPC-1 cells. The intensity of VEGF expression in CFPAC-1 also increased time-dependently with IL-6 treatment. In 32 surgical pancreatic cancer tissues, 19 (59%) stained positive for VEGF and 26 (87%) positive for IL-6 receptor beta subunit. The correlation between IL-6 receptors and VEGF was significant (p = 0.0002 and p = 0.0019) while less correlation was seen between IL-6 and VEGF (p = 0.1937). CONCLUSION: Our results suggest that IL-6 is likely to take part in VEGF expression in both paracrine and autocrine fashion in pancreatic cancer. Induction of VEGF seems to be regulated by the extent of the IL-6 receptor expression on cancer cells. PMID- 12553039 TI - Expression and distribution of human telomerase catalytic component, hTERT, in human breast tissues. AB - BACKGROUND: Activation of telomerase is essential in a variety of malignant tumors. However, the regulation of its activation and the expression of human telomerase reverse transcriptase (hTERT) are not fully understood in breast tissues. MATERIALS AND METHODS: In order to clarify this point, the telomerase activity was examined by telomeric repeat amplification protocol (TRAP) assay and quantified by hybridization protection assay (HPA) and, moreover, the expressions of hTERT mRNA and protein were examined by RT-PCR and immunohistochemistry, respectively. RESULTS: Telomerase activity and the expression of hTERT mRNA were detected in 41(75%) and 37(67%) cases among 55 breast carcinomas, respectively. However, the telomerase ZV activity was detected in only 14(40%) among 35 benign breast tumors. Strong correlation was observed between telomerase activity and hTERT mRNA expression, as described previously. Moreover, the strong expression of hTERT protein in the nucleus of the cell was detected in 30 (57%) cases among 53 breast carcinomas and weak expression of the protein was also detected in 11 (35%) cases among 31 benign breast tumors by immunohistochemistry. There were 12 breast carcinomas which had no telomerase activity. Three of them had hTERT mRNA and protein and in 3 other cases, only hTERT protein was detected. Five cases had the expression of mRNA but not the protein and it was only one case which had no activity and expression of hTERT mRNA and protein. However, in benign tumors, 14 (87.5%) cases among 16 TRAP-negative tumors had no mRNA and protein expression. CONCLUSION: These results indicate that complicated regulation might be involved in the regulation of telomerase activity and also the activation of the gene may be important in the carcinogenesis of human breast carcinomas. PMID- 12553040 TI - Mapping the methylation pattern by bisulfite genomic sequencing of the E-cadherin promoter CpG island in nasopharyngeal carcinoma. AB - BACKGROUND: It is hypothesised that hypermethylation of the CpG island in the promoter region in several tumour suppressor genes contributes one of the mechanisms for silencing or down-regulating the expression of the tumour suppressor genes. One of the putative tumour suppressor genes, the E-cadherin gene, has shown down-regulation in nasopharyngeal carcinoma (NPC). The aim of this study was to explore the methylation profiles of the E-cadherin promoter region in NPC patients. MATERIALS AND METHODS: Fifteen NPC paraffin-embedded tissue sections were included in the study accompanied by two cell lines, MCF7 and MDA435, as controls. The technique of bisulfite genomic sequencing was used to determine the methylation patterns of the 29 CpG dinucleotides along the E cadherin promoter region. DNAs were extracted from the samples, and then were subject to bisulfite conversion. Nested polymerase chain reactions were performed on the modified DNAs, the sequences of which were determined by direct sequencing. RESULTS: A pilot study ascertained the correlation between reduced E cadherin expression and hypermethylation of the E-cadherin promoter region from the E-cadherin (+) MCF7 and the E-cadherin (-) MDA435 cell lines. The experimental study revealed that, among 10 NPC cases in which DNA could be successfully extracted, 6 (60%) displayed methylation patterns at two or more CpG sites (range: 2-11, 6.8-37.9%). Five of them (83.3%) were comprised of at least 2 transcription enhancing factor sites. Two of the 4 cases (50%) with methylated 4th CpG site (HpaII site) and 3 of the 4 cases (75%) with methylated 11th CpG site (GC region) had grades 3 or 4 level of methylation. CONCLUSION: The data indicated that hypermethylation of the E-cadherin promoter region occurred in a large proportion of NPC patients, and frequent methylation of specific CpG sites associated with transcription enhancing elements suggested that this phenomenon may contribute part of the mechanisms for silencing or down-regulating E-cadherin gene expression in NPC. PMID- 12553041 TI - Induction of p57 is required for cell survival when exposed to green tea polyphenols. AB - Green tea polyphenols (catechins) are known to induce cell death in many types of tumor cells, but how normal epithelial cells survive in the presence of polyphenols is unknown. We recently reported that green tea polyphenols potently induced a cyclin-dependent kinase inhibitor, p57/(KIP2), only in normal human epithelial cells. In this study, we investigated the correlation between p57 expression and survival/apoptosis by Western blot analysis, caspase 3 assays and morphological analysis. It was demonstrated that, in the cells that lack p57 induction, green tea polyphenols induced Apaf-1 expression along with caspase 3 activation, leading to apoptosis. In contrast, cells with polyphenol-inducible p57 maintained constant levels of Apaf-1 and proliferating cell nuclear antigen (PCNA), with basal caspase 3 activity. Retroviral-transfected, p57-expressing oral carcinoma cells showed significant resistance to green tea polyphenol induced apoptosis. Our results suggest that p57/KIP2 is a determinant pro survival factor for cell protection from green tea polyphenol-induced apoptosis. PMID- 12553042 TI - The role of myoepithelial cells in the short-term radiogenic impairment of salivary glands. An immunohistochemical, ultrastructural and scintigraphic study. AB - BACKGROUND AND PURPOSE: The aim of this study was to evaluate the early dose related functional impairment of salivary glands after radiation, using sialoscintigraphy, immunohistochemistry and electron microscopy in an established rabbit experimental model. MATERIALS AND METHODS: Twelve rabbits were used for the study. Eight were scintigraphically examined prior to and 24 hours after 15/30 Gy (4 rabbits each). The irradiated glands were examined immunohistochemically using monoclonal antibodies to alpha-smooth muscle actin (ASMA), vimentin and Ki-67 proliferation antigen. Ultrastructural investigation was also performed. Four control rabbits were sham-treated and provided normal salivary gland tissue. RESULTS: There was a significant increase of the 99mTc pertechnetate uptake in the irradiated parotid glands (p < 0.05) and a highly significant one in the superficial mandibular glands (p < 0.001). Immunohistochemically a significant loss of ASMA and vimentin-stained myoepthelial cells and a decrease of the proliferating rate in the acinar cells could be assessed in both irradiated glands. Ultrastructurally, rarefaction and focal condensation of the myofilaments of the myoepithelial cells in both irradiated glands was observed. No dose-related effect could be assessed. CONCLUSION: Early functional impairment of salivary glands after radiation could be revealed as early as 24 hours after radiation. The considerable myoepthelial cell impairment may explain the secretory retention assessed scintigraphically and provide -apart from acinar cell damage--a new aspect in the pathogenesis of radiogenic impairment of salivary glands. PMID- 12553043 TI - Anti-metastatic activities of all-trans retinoic acid, indole-3-carbinol and (+) catechin in Dunning rat invasive prostate adenocarcinoma cells. AB - Dunning rat invasive prostate adenocarcinoma cells were employed to investigate the anti-metastatic potential and probable mechanisms of action of all-trans retinoic acid (ATRA), indole-3-carbinol (13C) and (+)-catechin (CAT). The invasive parameters studied include: matrigel membrane invasion; zymography and Northern analysis for matrix metalloproteinases (MMPs) activity and gene expression; and Western analysis for the membrane-associated proteins alpha-, beta- and gamma-catenins. ATRA significantly and dose-dependently inhibited matrigel membrane invasion of the cells by 53%, inhibited MMP-2 activity by 71%, MMP-9(80%), alpha-(59%) and beta-(65%) catenin expression at 10 microM (p < 0.01). gamma-Catenin expression was completely inhibited by ATRA even at 2 microM. Catechin at 25 microM decreased matrigel membrane invasion by 24% and also inhibited gamma-catenin protein levels by 58% (p < 0.01). Loss of E-cadherin was implicated in the exacerbation of the anti-metastatic effects of ATRA and CAT by the use of E-cadherin-positive, non-invasive cells. In conclusion, ATRA and CAT show anti-metastatic potential in the invasive rat prostate adenocarcinoma model and gamma-catenin appears to play a mechanistic role. PMID- 12553044 TI - Hypoxia inducible factor 1 alpha and 2 alpha expression is independent of anemia in patients with stage I endometrial cancer. AB - OBJECTIVE: To investigate whether anemia, a putative factor counteracting the efficacy of radiotherapy, up-regulates the endogenous markers of intratumoral hypoxia, the hypoxia inducible factors HIF1 alpha and HIF2 alpha. MATERIALS AND METHODS: Forty-two hysterectomy specimens harboring endometrial adenocarcinomas of the endometrioid cell type, stage I were investigated immunohistochemically for the expression of HIF1 alpha, HIF2 alpha and the down-stream inducible expression of vascular endothelial growth factor VEGF. The results were correlated with hemoglobin (Hb) levels. RESULTS: There was no significant association between Hb levels and the expression of HIF1 alpha, HIF2 alpha or VEGF in our material. CONCLUSION: Activation of hypoxia pathways is an intrinsic self-regulated process, independent of Hb levels in endometrial adenocarcinomas. Other factors such as microvessel density (MVD), vessel/tumor cell distance or genetic events may be important. PMID- 12553045 TI - The cytotoxicity of a conjugate composed of human epidermal growth factor and eosinophil cationic protein. AB - BACKGROUND: Conventional targeted therapy with foreign proteins is highly immunogenic. In this study, we developed targeted therapy composed of human endogenous proteins and evaluated its efficacy in vitro. MATERIALS AND METHODS: Human epidermal growth factor (EGF) was chemically linked to human eosinophil cationic protein (ECP). The cytotoxicity of the EGF-ECP conjugate was evaluated by MTT assay. RESULTS: The conjugate showed dose-dependent cytotoxicity on EGF receptor (EGFR)-overexpressing BT-20 cells with an IC50 of 1.5 x 10(-7) M, whereas the IC50 of ECP alone was almost 10(-4) M. The conjugate had no detectable cytotoxicity against EGF receptor-deficient H69 cells. Excess EGF protected BT-20 cells from the cytotoxicity of the conjugate. Comparing the cytotoxicity and the level of EGFR expression, the cytotoxicity of the conjugate was positively correlated with the level of EGFR expression of each cell line. CONCLUSION: Conjugates composed solely of human proteins might be useful with less immunogenicity and less toxicity than the conventional immunotoxins for targeted therapy. PMID- 12553046 TI - The suitability of paraffin-embedded material to predict metastatic potential of oral squamous cell carcinoma. AB - BACKGROUND: The presence of regional metastasis in oral squamous cell carcinoma (OSCC) is an important prognostic factor. This study was undertaken to identify histological features and biological markers from paraffin-embedded primary OSCC that may predict the presence of regional metastases. MATERIALS AND METHODS: Fifty-three en-bloc primary OSCC resections were divided into two groups, 26 with lymph node metastases and 27 without metastases. The pattern of infiltration, presence of vascular or perineural infiltration and tumour necrosis were evaluated while expression of p53, p21 and Rb were assessed in the two groups. DNA ploidy status was also determined with a flow cytometer. RESULTS: The presence of DNA aneuploidy was found to be the only statistically significant predictor of regional metastases. Seventy-seven per cent of the primary OSCC with lymph node metastases showed DNA aneurploidy. CONCLUSION: DNA flow cytometry obtained from archival material could be used as a parameter to predict regional metastases. PMID- 12553047 TI - Inhibition of murine osteosarcoma cell proliferation by glucocorticoid. AB - The effects of glucocorticoid (GC) on the proliferation of Dunn Osteosarcoma (OS) cells were examined under in vitro culture conditions. Dexamethasone (Dex) inhibited the proliferation of Dunn OS cells in a dose-dependent manner, while the addition of anti-GC, RU486, to the culture medium in part recovered Dex induced growth inhibition. The number of maximum binding sites (Bmax) and the dissociation constant (Kd) value of glucocorticoid receptor (GR) in Dunn OS cells were 19,560 sites/cell and 5.2 +/- 0.8 nM, respectively. RU486 competed with labeled Dex against GR at a concentration of 10(-6) M. Western blot analysis of [3H]Dex-mesylate-labeled cell homogenate and immunohistochemical staining against GR further confirmed the presence of GR. Dex treatment of Dunn OS cells resulted in apoptosis with the characteristic internucleosomal DNA cleavage shown by the DNA ladder pattern in agarose gel electrophoresis. These data demonstrate that GC inhibits the proliferation of Dunn OS cells via GR, for which one possible mechanism in vitro is induction of apoptosis. PMID- 12553048 TI - Synthesis and expression of mRNA encoding for different versican splice variants is related to the aggregation of human epithelial mesothelioma cells. AB - Malignant mesothelioma often has a biphasic growth pattern of epithelial and/or sarcomatous morphology. In culture, epithelial cells form aggregates, whereas fibroblast-like cells do not. Two human mesothelioma cell sub-lines, one with epithelial differentiation and the other with fibroblast-like phenotype were studied. We have previously shown (Dobra et al, 2000) that distinct types of the cell-associated syndecans are involved in the regulation of mesothelioma cell differentiation, whereas the role of matrix proteoglycans (PGs) remains unknown. This study was undertaken to examine whether cell aggregation of the epithelial mesothelioma cells correlates to the differential expression of the matrix PGs versican and perlecan at different degrees of confluence. PGs were isolated from the culture medium using ion-exchange chromatography and identified by high performance liquid chromatography, capillary electrophoresis and Western blotting. Fibroblast-like cells express substantially more versican than epithelial cells. RT-PCR showed that both cell lines express mRNA coding for versican splice variants V0 and V1, but not for V2. The dominating splice variant in both cell lines is the V0. Screening of versican splice variants in various degrees of culture confluence showed that the expression of mRNA conding for the versican splice variants V0 and V1 is different only in confluent cultures. No significant differences in the expression of perlecan between the two cell lines were recorded. These results suggest that the aggregation of epithelial cells is related to a significant decrease (p < or = 0.001) of the splice variant V1. This variant seems to be a biologically active constituent that affects tumor biology. PMID- 12553049 TI - A proapoptotic caspase recruitment domain protein gene, TMS1, is hypermethylated in human breast and gastric cancers. AB - BACKGROUND: Conway et al. demonstrated that methylation of the proapoptotic gene, TMS1, was observed in breast cancer cell lines and tissues, resulting in decreased TMS1 gene transcription. However, whether the TMS1 gene is hypermethylated in other cancers is uncertain. MATERIALS AND METHODS: The expression of TMS1 mRNA was determined by quantitative RT-PCR. Methylation of the TMS1 gene was detected using methylation-specific PCR followed by bisulfite modification of DNA. RESULTS: Methylation of the TMS1 gene was observed in breast, gastric and colorectal cancer cells. Down-regulation of TMS1 gene transcription in colorectal cancer cells was restored by treatment with a demethylating agent. Methylation of the TMS1 gene was observed in 2 out of 19 breast cancer specimens and 1 out of 9 gastric cancers, but in none of 13 colorectal cancers. CONCLUSION: These results suggest a direct role for aberrant methylation of the TMS1 gene in the progression of breast and gastric cancer involving down-regulation of the proapoptotic TMS1 gene. PMID- 12553050 TI - Differential expression of aggrecan mRNA isoforms by chondrosarcoma cells. AB - BACKGROUND: Aggrecan, the large aggregating proteoglycan, is a major component of the cartilage extracellular matrix. The C-terminal globular domain (G3) of the aggrecan molecule exhibits a pattern of alternative splicing that involves three exons. Each corresponds to a subdomain, epidermal growth factor 1(EGF1), EGF2, and a short complement repeat (SCR). MATERIALS AND METHODS: In the present study, we compared aggrecan mRNA splicing variants in osteochondrogenic tumor cells and normal cartilage using RT-PCR. RESULTS: Normal cartilage expressed two mRNA isoforms, both containing the SCR exon, but with or without the EGF1 exon. Benign cartilaginous tumors showed the same splicing pattern as normal cartilage. However, malignant chondrosarcomas expressed additional splicing isoforms, one of which lacked all three alternative exons and another with EGF2 and SCR exons. The human chondrosarcoma cell line (Ch-1 cells) expressed further isoforms, one containing either EGF1 or EGF2 exon only, and another with all the variant exons. CONCLUSION: These results imply that a change in the pattern of alternative aggrecan mRNA splicing is associated with malignant transformation of chondrocytes. PMID- 12553051 TI - Selection of an internal control gene for quantitation of mRNA in colonic tissues. AB - BACKGROUND: GAPDH, beta-actin and 18S rRNA are widely employed as internal control genes, with the assumption that they are expressed constitutively to similar degrees in different cells and tissues and under different experimental conditions. In this study, we tested this assumption by assessment of the transcription of these three genes in human colonic tissues using a quantitative RT-PCR. RESULTS: GAPDH transcription was significantly greater in both colonic adenomas and cancers than in normal mucosa. In addition, transcription of beta actin was significantly increased in cancers. The expression of 18S rRNA was essentially constant among these various tissues. Stable expression of 18S rRNA was observed during the growth of colonic cancer cells stimulated with serum, but both GAPDH and beta-actin transcription were up-regulated, coinciding with cell proliferation. CONCLUSION: These results indicate that 18S rRNA is more reliable than GAPDH and beta-actin as an internal control gene for quantitative comparison of mRNA in colonic cancers. PMID- 12553052 TI - Turmeric (Curcuma longa) and curcumin inhibit the growth of Helicobacter pylori, a group 1 carcinogen. AB - BACKGROUND: Curcumin, a polyphenolic chemical constituent derived from turmeric (Curcuma longa), has been shown to prevent gastric and colon cancers in rodents. Many mechanisms have been proposed for the chemopreventative effects, although the effect of curcumin on the growth of Helicobacter pylori has not been reported. H. pylori is a Group 1 carcinogen and is associated with the development of gastric and colon cancer. MATERIALS AND METHODS: A methanol extract of the dried powdered turmeric rhizome and curcumin were tested against 19 strains of H. pylori, including 5 cagA+ strains. RESULTS: Both the methanol extract and curcumin inhibited the growth of all strains of H. pylori in vitro with a minimum inhibitory concentration range of 6.25-50 micrograms/ml. CONCLUSION: These data demonstrate that curcumin inhibits the growth of H. pylori cagA+ strains in vitro, and this may be one of the mechanisms by which curcumin exerts its chemopreventative effects. PMID- 12553053 TI - A novel gene containing PDZ and LIM domains, PCD1, is overexpressed in human colorectal cancer. AB - BACKGROUND: The process of colorectal cancer development involves accumulated genetic alterations affecting APC, K-ras and p53. A recently identified gene, PCD1, was reported to be up-regulated in human malignancies including colorectal cancers, but relationships between PCD1 gene expression and clinicopathological findings, as well as the timing of genetic alteration of PCD1 in colorectal cancer development, are not clear. To determine whether PCD1 contributes to colorectal cancer progression, we investigated the expression of PCD1 mRNA in human colorectal tissues. MATERIALS AND METHODS: The expression of PCD1 mRNA was determined by quantitative RT-PCR. The mutation of p53 was detected by a PCR-SSCP method. RESULTS: Up-regulation of PCD1 gene transcription was observed not in adenomas but in cancers compared to normal mucosa (p < 0.0001). Primary tumors with a mutation of p53 showed significantly greater PCD1 gene expression than tumors without such a mutation (p = 0.0134). CONCLUSION: The PCD1 gene may play a role in colorectal cancer development from adenomas. PMID- 12553054 TI - Standardized mistletoe extract augments immune response and down-regulates local and metastatic tumor growth in murine models. AB - The immunomodulatory and antimetastatic activity of standardized aqueous mistletoe extract (sME) was evaluated in BALB/c-mice. Regular subcutaneous (s.c.) applications (three times per week for 14 consecutive days; 2, 20, 100 and 500 micrograms per injection and mouse) up-regulated thymocyte and peripheral blood leukocyte counts in tumor-bearing mice. Tumor weight and tumor volume were significantly down-regulated after application of sME doses greater than 20 micrograms per injection. To check the influence of sME treatment on growth of experimental metastases, RAW 117 H 10 lymphosarcoma cells and L-1 sarcoma cells were intravenously inoculated into BALB/c-mice to establish liver and lung colonization, respectively. sME was regularly administered starting 24 hours after tumor cell challenge. Organ colonization was investigated on day 14 after tumor cell inoculation and demonstrated statistically significant (p < 0.05) reductions of experimental liver and lung metastases for sME-treated mice. PMID- 12553055 TI - Cytotoxic activity of a new lipid formulation of doxorubicin in cell lines and primary tumor cells. AB - BACKGROUND: Liposomal formulations of the anthracyclines are being developed to circumvent toxicity and prolong effect. The current study investigates the in vitro activity of a novel doxorubicin micelle formulation, containing a vehicle designed to release pharmacologically active subcomponents. MATERIALS AND METHODS: The cytotoxicity of doxorubicin formulated in a vehicle containing C4 (N docosahexaenoyl-O-phospho-2-aminoethanol) and C11 (N-all trans-retinoyl-O-phospho L-tyrosine) was measured in a panel of human tumor cell lines, 19 primary cultures of human tumor cells and 5 lymphocyte preparations. RESULTS AND CONCLUSION: At the tested ratio between doxorubicin and C4/C11 (1:50), C4/C11 contributed significantly to the in vitro toxicity. However, the molar EC50 values were lower for doxorubicin than for C4/C11. Synergistic interactions between doxorubicin and C4/C11 were evident in a majority of the cell types studied. C4/C11 increased the cellular load of the fluorescent Pgp substrate calcein. To further investigate the possible benefits of the new formulation, in vivo studies are ongoing. PMID- 12553056 TI - Circulating nerve growth factor level changes during oxaliplatin treatment induced neurotoxicity in the rat. AB - BACKGROUND: Oxaliplatin neurotoxicity represents a clinically-relevant problem and its etio-pathogenesis is still unknown. We explored the possible role of some neuronal growth factors ("neurotrophins") during the course of oxaliplatin sensory neuronopathy. MATERIALS AND METHODS: In our rat model two different doses of oxaliplatin were used (2 and 3 mg/kg i.v. twice weekly for 9 times). The neurotoxicity of the treatment was assessed with neurophysiological and pathological methods and serum neurotrophin levels were measured by ELISA. RESULTS: Both oxaliplatin-treated groups showed the neurophysiological and neuropathological changes which mimic the chronic effects of oxaliplatin administration in humans, e.g. reversible sensory impairment due to dorsal root ganglia neuron damage. These changes were associated with a significant and dose dependent reduction only in the circulating level of nerve growth factor (NGF), which returned to normal values after neurophysiological and pathological recovery. CONCLUSION: This specific association between neurological impairment and NGF modulation indicates that NGF impairment has a role in the neurotoxicity of oxaliplatin. PMID- 12553057 TI - Induction of N-myc in neuroblastoma by autocrine IGF-II depends on farnesylated Ras. Application of farnesyltransferase inhibitors. AB - Genetic aberrations are the primary events leading to carcinogenesis in various tissues and are characteristic for certain tumor types. Amplification of N-myc and deletion of 1p significantly correlate with poor prognosis of neuroblastoma patients. Very little informations is available on the regulation of N-myc expression by external factors. Insulin-like growth factor-II (IGF-II) has been identified as an autocrine growth factor in neuroblastoma. Four neuroblastoma cell lines were examined for their expression of IGF-II and IGF-receptor. Stimulation of neuroblastoma cells with IGF-II leads to an increased activity of the MAP-kinase Erk1, an induction of N-myc expression and an enhanced proliferation rate. In order to disrupt the signal transduction of the IGF receptor, we inactivated the Ras-proteins in neuroblastoma cells by inhibition of the farnesyl-protein transferase by FTI-277. This inactivation prevented activation of MAP-kinase Erk1 and induction of N-myc expression by IGF-II. Inactivation of Ras by farnesyltransferase inhibitors might become a promising new approach in future treatments of neuroblastoma tumors. PMID- 12553059 TI - Stimulation of the maturation of dendritic cells in vitro by a fermented mistletoe extract. AB - BACKGROUND: Dendritic cells (DC) play a key role during the initiation of specific immune responses. In cancer patients, however, an alteration of their function was observed. In our investigation we analysed the influence of a fermented mistletoe extract often used for adjuvant treatment of cancer patients on the generation and maturation of DC. MATERIALS AND METHODS: Monocytes from healthy individuals were incubated with a fermented mistletoe extract in the presence or absence of GM-CSF/IL-4. Surface marker expression was measured by flow cytometry. RESULTS: While there was no relevant effect on the generation of DC in the absence or presence of GM-CSF/IL-4 in 5-day cultures, the mistletoe extract significantly stimulated the maturation of pre-generated immature DC, as evidenced by a heightened expression of CD83. Like the positive control TNF alpha, the mistletoe extract significantly activated CD80 and CD86 as well as HLA class I and II molecules on these cells. CONCLUSION: Our data clearly demonstrate an influence of the mistletoe extract on the maturation of DC, but it remains to be elucidated whether the function of DC is also activated and, especially, whether this effect can be observed in tumour patients as well. PMID- 12553058 TI - Notch signal transduction is not regulated by SEL1L in leukaemia and lymphoma cells in culture. AB - The transduction of Notch signal plays an intricate role in cell differentiation and pathogenesis of haematological malignancies as well as in certain congenital conditions. The functionality of Notch signalling was tested using HES1 gene activation. SEL1 gene product has been postulated to be a negative regulator of Notch signalling. We investigated the relationship between Notch signalling and the expression of SEL1L gene in a number of leukaemia and lymphoma cells in culture. The cell lines could be separated into two groups. Group 1 contained lymphoma cell lines in which Notch signalling was intact; of these 4 cell lines were SEL1L+/HES1- and 3 SEL1L-/HES1-. Notch signalling was not subverted by EBNA2 expression in these lymphoma cells. In Group 2 cell lines Notch signalling was constitutively active but 6 out of 7 cell lines expressed SEL1L at high levels. In summary, a majority of cell lines of both groups express SEL1L and no inverse relationship is evident between SEL1L expression and the status of Notch signalling. The present investigation therefore suggests that SEL1L may not exert a negative regulatory influence on Notch signalling. No genomic alterations affecting SEL1L were detected either in the lymphoma or T-ALL cell lines tested. Taken together the present findings do not support the postulated negative regulatory role for SEL1L in Notch signalling. PMID- 12553060 TI - Activity of ascaridol from the anthelmintic herb Chenopodium anthelminticum L. against sensitive and multidrug-resistant tumor cells. AB - Ascaridol is the active principle of the American wormseed Chenopodium anthelminticum L. We isolated ascaridol from a commercial preparation of Chenopodium oil and analyzed its activity against different tumor cell lines in vitro (CCRF-CEM, HL60, MDA-MB-231). Multidrug-resistant (MDR) counterparts of these cell lines express differentially the MDR-conferring ATP-binding cassette transporter genes MDR1, MRP1 and BCRP, respectively. We found that ascaridol exerts antineoplastic activity. The findings of the present investigation are the first hint that ascaridol may be an interesting novel candidate drug for cancer treatment. PMID- 12553061 TI - In vitro antiangiogenic effect of sandostatin (octreotide) on the proliferation of the placental vessels. AB - BACKGROUND: The somatostatin analog octreotide (Sandostatin) exerts its antineoplastic effect through different mechanisms. There are several in vitro and in vivo studies available demonstrating an antiangiogenic activity, however, some other observations failed to reinforce these results. We investigated the potential angio-inhibiting activity of this drug in an in vitro system using human placental fragments. MATERIALS AND METHODS: In an in vitro angiogenesis assay, small pieces of human placenta were embedded in a fibrin gel and the effect of 1, 10 and 100 micrograms/ml Sandostatin was assessed on the microvessel formation. The results were expressed as microvessel counts per mm perimeter. RESULTS: At a dose of 1 microgram/mL octreotide did not influence the microvessel outgrowth, however, a 10 micrograms/ml concentration resulted in a 61% inhibition, while in the samples treated with 100 micrograms/ml Sandostatin only occasional capillary tubes formed. CONCLUSION: Sandostatin effectively inhibited the outgrowth of the placental vessels in a dose-dependent manner. In the mechanism of action of this drug the antiangiogenic effect should also be taken into account. PMID- 12553062 TI - Induction of drug resistance and protein kinase C genes in A2780 ovarian cancer cells after incubation with antineoplastic agents at sublethal concentrations. AB - We examined the inducibility of drug resistance (MDR1, MRP1, LRP) and protein kinase C (PKC) isozyme (alpha, epsilon, eta, theta, tau, zeta) corresponding genes in A2780 ovarian cancer cells after a 24-hour treatment with adriamycin (ADR), camptothecin (CAM), etoposide (ETO) or vincristine (VCR). Sublethal concentrations of drugs were used to exclude short-term effects caused by selection. Cell cycle analysis was performed to identify possible correlation between resistance factors, PKC isozymes and proliferation. We found a mostly combined induction of MDR1, LRP, PKC tau and PKC zeta by CAM, ETO and VCR. PKC alpha, epsilon, eta and theta gene expression altered variably. Cell cycle analysis showed that A2780 cells responded with a marked G2/M arrest after a 24 hour treatment with CAM, ETO and VCR but an association between the induction of PKC isozymes corresponding genes and proliferation was not seen. Our analysis points to a possible link between atypical PKC tau/PKC zeta and MDR1/LRP in cytostatic stress response of cancer cells. PMID- 12553063 TI - In vitro immunomodulatory effects of cuphiin D1 on human mononuclear cells. AB - Cuphiin D1 (CD1), a macrocyclic hydrolyzable tannin isolated from Cuphea hyssopifolia, has been shown to exert antitumor activity both in vitro and in vivo. Moreover, the antitumor effects of CD1 are not only related to its cytotoxicity to carcinoma cell lines, but also depend on host-mediated mechanisms. In the present study, CD1 was investigated for its effects on the proliferation and cytokine secretion of human peripheral blood mononuclear cells (PBMCs). At concentrations of from 6.25 to 50 micrograms/ml, it enhanced the 3H thymidine incorporation of concanavalin A (Con A)-stimulated PBMCs in a dose dependent manner. Excretion of IL-1 beta, IL-2 and TNF-alpha by CD1-stimulated PBMCs was markedly increased in a dose-dependent manner. The results show that CD1 could stimulate PBMCs release of IL-1 beta, IL-2 and TNF-alpha and then activate T cells. Therefore, CD1-activated T cells via IL-1 beta in vitro might account for the host-mediated CD1 mechanism of action. PMID- 12553064 TI - The complement inhibitor CD59 and the lymphocyte function-associated antigen-3 (LFA-3, CD58) genes possess functional binding sites for the p53 tumor suppressor protein. AB - p53 is an oncosuppressor protein, which acts via transcriptional and non transcriptional mechanisms. The transcriptional function of p53 is mediated by specific responsive elements. In the present study we found active responsive elements, specific for the p53 within the 5'flanking region and within the first intron of the gene encoding for the CD59 membrane inhibitor of reactive lysis, and within the first intron of the gene encoding for the CD58 membrane protein (LFA-3). The results suggest that p53 may enhance the transcription of both CD59 and CD58 and imply a novel role for p53 as a direct regulator of the immune response. PMID- 12553065 TI - Mechanisms of hyperthermia- and 4-hydroperoxy-ifosfamide-induced cytotoxicity in T cell leukemia. AB - The prognosis of patients with early ALL (acute lymphoblastic leukaemia) relapse is poor with conventional chemotherapy alone. Thus, intensified chemotherapy strategies are required. The application of hyperthermia enhances the efficacy of certain antineoplastic drugs such as ifosfamide. In this study, the effects and molecular mechanisms of ifosfamide (4hydroperoxy-ifosfamide = 4OOH-IFA)- and/or hyperthermia-induced cell death are investigated in CEM cells. Hyperthermia enhanced the efficacy of 4OOH-IFA in a subaddictive manner. Analysis of caspase activation revealed an early hyperthermia-induced stimulation of caspase-3 and -6 directly after the heating pulse, while maximum activation following stimulation with 4OOH-IFA was obtained after 24 hours of culture. The combination of 4OOH-IFA and hyperthermia mediated an overaddictive caspase stimulation directly following the heating phase. At this time also an overaddictive cytotoxic effect was noticed, being mainly responsible for the enhancing effects of hyperthermia on 4OOH-IFA cytotoxicity. In conclusion, hyperthermia enhanced the cytotoxic effect of 4OOH-IFA on CEM cells by stimulation of an early 4OOH-IFA effect. Thus, thermochemotherapy might be considered as an intensifying treatment option in relapsed T cell leukemias. PMID- 12553066 TI - Significance of tumor-associated antigens in the diagnosis and therapy of cancer: an overview. AB - An enormous effort using a wide variety of approaches has been undertaken over the last three decades to transform both basic and clinical research into improved diagnoses and therapies of cancer. This brief overview summarizes the significance of tumor-associated antigens (TAAs) in the diagnosis and therapy of cancer. Current data suggest that immunotherapy and gene therapy using antibody recognized TAAs as their targets are promising, whereas those using T cell recognized peptide epitopes of TAAs as their targets remain controversial regarding their efficacy, mainly due to general losses of HLA molecules in tumor cells. PMID- 12553067 TI - The role of proteolysis in tumor invasiveness in glioblastoma and metastatic brain tumors. AB - Local invasive infiltration and growth are key features in glioblastoma, which are accompanied by remodeling of the vasculature and the destruction of the surrounding normal brain tissues. The local and regional spread of glioblastoma is often associated with poor prognosis. The invasive character of glioblastoma appears to depend partly on the proteolytic destruction of the extracellular matrix components. In this article, we review the role of proteolytic enzymes, urokinase-type plasminogen activator and matrix-metalloproteases, for diffuse infiltrative growth of glioblastoma. An understanding of the mechanisms of increased expression of these molecules in glioblastoma might provide an insight into the regulatory pathway of cellular invasion and for a new therapeutic strategy for glioblastoma. PMID- 12553068 TI - Role of tumor-associated macrophage in malignant tumors: should the location of the infiltrated macrophages be taken into account during evaluation? AB - Tumor-associated macrophages (TAMs) have a complex relationship with the tumor cells. Because TAMs have various functions according to the microenvironment, the results of many analyses have not led to a general consensus regarding the role of TAMs in solid tumor tissue and the prognostic significance for patients. In this review, we discuss whether the recently published data can help to explain the apparent paradoxical effects of TAMs in cancers. We address this question on the basis of the histological location of TAMs in solid malignant tumors. PMID- 12553069 TI - Perspectives for antitumor therapy based on the recognition mechanism by which macrophages identify undesirable foreign and native cells and molecules. AB - Though immunotherapy appears to be a promising therapy for tumors with antigenicity, it may have only a limited usefulness, because tumor cells do not reveal tumor-specific antigens. In contrast, macrophages may be able to distinguish and remove tumor cells because they can recognize and remove both undesirable foreign and native substances, and can do this independently of MHC or antigenicity. For the development of a new antitumor therapy based on the molecular mechanism to enhance the elimination of tumor cells, it is considered important to clarify the recognition mechanism of undesirable foreign and native substances by macrophages. Advanced data in this field of research have been rapidly accumulated recently. The macrophage recognition system for undesirable foreign and native substances is based on multiple receptors which bind to various kinds of foreign substances, suggesting a complicated mechanism. Much of this system is still undescribed and further study is needed. Even in the most advanced research areas (the recognition system of bacteria and their products by macrophages), the data still suggest the existence of unknown receptors. We therefore believe that research into the nature of these unknown receptors can provide information which may be used to develop novel antitumor therapies in the future. PMID- 12553070 TI - Clinical effects of tumor-associated macrophages and dendritic cells on renal cell carcinoma. AB - Several reports have documented infiltration of many monocytes in renal cell carcinoma (RCC). Since few studies have examined the relationship between monocyte infiltration and clinical prognosis in RCC, we clinically investigated this relationship by semi-quantitative analysis of monocyte infiltration and tumor angiogenesis. The following six parameters were measured immunohistologically in 98 RCC patients who underwent nephrectomy between 1987 and 1997: tumor-associated macrophage (TAM), microvessel density (MVD), S-100 protein-positive cells (S-100(+) cells), HLA-DR-positive cells, apoptosis index and proliferative index (PI). We then assessed intercorrelations among parameters and correlations to prognosis. Significant positive correlations were identified for TAM, MVD and PI, with a tendency for higher parameter values to reflect poorer prognosis. The prognosis of patients without metastasis was poor for the high TAM group even when levels of MVD were low. These findings suggest that TAM facilitates the growth of RCC via angiogenesis and other mechanisms. Prognosis was significantly better in metastatic RCC patients who underwent interferon alpha (IFN-alpha) therapy when the levels of S-100(+) cells were high. Nonetheless, the levels of S-100(+) cells among these IFN-treated patients did not correlate with other parameters, and none of the other parameters correlated with prognosis. One of the antitumor effects of IFN-alpha for RCC could therefore be mediated by dendritic cells. PMID- 12553072 TI - Relationships between tumor-associated macrophages and clinicopathological factors in patients with colorectal cancer. AB - BACKGROUND: As macrophages have both tumor promoting and inhibiting effects, tumor-associated macrophages (TAM) play a complex role in the regulation of cancer progression and tumor growth. The clinicopathological significance of TAMs in human colorectal cancer remains unclear. MATERIALS AND METHODS: We investigated the relationships between TAM IF (invasive front), TAM TS (tumor stroma), prognosis and clinicopathological factors in 30 patients with colorectal cancer. Moreover, MVD (microvessel density) was examined for correlation with these factors. TAMs were detected by immunohistochemistry using the anti-CD68 antigen, KP1 mouse monoclonal antibody. MVD was determined by counting the number of CD34-positive vessels. RESULTS: Low TAM IF and TAM TS tended to occur with more advanced colorectal cancer, although this finding was not statistically significant. High TAM IF tended to be found in patients with a good prognosis or those with less recurrence or worsening of their condition. MVD was significantly higher in patients with lymph node metastasis, lymphatic invasion or more advanced tumors than stage III (TNM). Although the correlation between TAM IF, TAM TS and MVD was not statistically significant, the correlation between TAM IF and TAM TS was statistically significant. CONCLUSION: These results suggest that TAMs may act as one line of defence against malignant cells in patients with colorectal cancer. PMID- 12553071 TI - Targeting of T cells to CEA-expressing tumor cells by chimeric immune receptors with a highly specific single-chain anti-CEA activity. AB - BACKGROUND: Lymphokine-activated killer (LAK) cell therapy or cytokine gene therapy is a potential new modality in the treatment of cancer. However, one limitation in the use of adoptive cellular immunotherapy for cancer lies in the difficulty in obtaining tumor-specific effector cells which can efficiently reach their target cells. MATERIALS AND METHODS: To effectively target T cells to carcinoembryonic antigen (CEA)-positive tumor cells, we constructed a chimeric immune receptor (CIR) gene which encoded a highly specific anti-CEA scFv antibody, the human CD8 alpha hinge region, the CD28 transmembrane and cytoplasmic domains, and the human CD3 zeta-chain. The resulting CIR gene, termed F39scFv/CIR-2, was transfected into human T cells. RESULTS: The cell surface expression of the F39scFv/CIR-2 receptors in T cells was demonstrated by flow cytometry. When incubated with CEA-positive tumor cells, the transfected T cells formed rosette-like aggregates around the tumor cells, thus indicating the cell specific targeting of T cells. CONCLUSION: This strategy may allow the development of new approaches for the adoptive immunotherapy of CEA-positive tumors in humans, especially in combination with LAK cell therapy or cytokine gene therapy. PMID- 12553073 TI - Preparation of Gc protein-derived macrophage activating factor (GcMAF) and its structural characterization and biological activities. AB - BACKGROUND: Gc protein has been reported to be a precursor of Gc protein-derived macrophage activation factor (GcMAF) in the inflammation-primed macrophage activation cascade. An inducible beta-galactosidase of B cells and neuraminidase of T cells convert Gc protein to GcMAF. MATERIALS AND METHODS: Gc protein from human serum was purified using 25(OH)D3 affinity column chromatography and modified to GcMAF using immobilized glycosidases (beta-galactosidase and neuraminidase) The sugar moiety structure of GcMAF was characterized by lectin blotting by Helix pomatia agglutinin. The biological activities of GcMAF were evaluated by a superoxide generation assay and a phagocytosis assay. RESULTS: We successfully purified Gc protein from human serum. GcMAF was detected by lectin blotting and showed a high biological activity. CONCLUSION: Our results support the importance of the terminal N-acetylgalactosamine moiety in the GcMAF-mediated macrophage activation cascade, and the existence of constitutive GcMAF in human serum. These preliminary data are important for designing small molecular GcMAF mimics. PMID- 12553074 TI - Bone marrow-derived factors support growth of N-type, but not of melanocytic neuroblastoma cells. AB - BACKGROUND: Neuroblastoma and melanoma cells have a common embryonal origin. In contrast to melanoma, most neuroblastoma tumours preferentially metastasize into bone marrow. Previously, we described that bone marrow-conditioned medium (BM-CM) supports the proliferation of catecholamine-producing (N-type) neuroblastoma (SK N-SH, IMR-32, Kelly)-, but not of melanoma cells. Both neuroblastoma and melanoma produce DOPA (3,4 dihydroxyphenylalanine); while melanoma cells use tyrosinase for DOPA synthesis, neuroblastoma cells usually utilize tyrosine hydroxylase. RESULTS: Certain neuroblastoma cells (in our study: SK-N-LO, LS, SH-EP) express tyrosinase instead of tyrosine hydroxylase for synthesis of DOPA, and do not synthesize catecholamines, as shown by HPLC and RT-PCR analysis. Strikingly and in contrast to catecholamine-producing N-type cells, the proliferation of these melanocytic neuroblastoma cells is not supported by BM-CM. CONCLUSION: With respect to proliferation in the presence of BM-CM, melanocytic neuroblastoma cells behave more like melanoma cells and may represent the subfraction of neuroblastoma cells with a minor tendency to metastasize into bone marrow. PMID- 12553075 TI - Annual recertification program for audit standards used in the EPA PM2.5 Performance Evaluation Program. AB - This paper describes procedures used to perform 152 annual recertifications of temperature, pressure, and flow rate audit standards. It discusses the metrology laboratories and the uncertainty of their recertifications. It describes the data base for the standards that tracks their recertifications and shipments. Finally, it presents some illustrative recertification results and describes what these results reveal about the audit standards and the recertifications. PMID- 12553076 TI - Building the airplane in flight: an auditing approach to quality management system development. AB - In June of 2000, Maine DEP (in company with the other five New England states) found itself under EPA pressure to develop and document a quality management system by the end of the calendar year. In the frenzy that followed, the Department decided to use a private sector model for system development that called for a prospectively-focused QM plan that would be implemented through formal auditing. That is, instead of trying in advance to identify all the deficits in departmental quality management, and then assigning individuals and task groups to create structures to fill the gaps, Maine's QMP specifies the future desired system in broad terms. ME-DEP then uses its own cadre of trained auditors to assess current practice against the "condition expected" in the QMP, knowing that in many cases basic quality management practices will need to be developed. This approach assists program managers, particularly (but not exclusively) in areas sensitive to environmental data standards, in creating systems and practices that are rooted in reality, and that are perceived to add value to the Department's core work instead of just additional paperwork. Audit reports create a continuous feedback loop assuring that written procedures document actual operations. Finally, the results of auditing identify areas in which the QM system (and its plan) should be elaborated or refined, which leads to an iterative process by which quality approaches are infused in all areas of DEP operations. In the twelve months following EPA-Region I's initial approval of Maine's QMP, a total of seven audits were completed at various program levels, including two focused on critical QMP elements: Documents and Records; and Computer Hardware/Software. This paper will briefly describe the background and implementation of this approach; identify some of the factors which led to success; and describe, using selected examples, some of the early outcomes of the program. PMID- 12553077 TI - Adventures in environmental data reporting: high tech, low tech, and everything in between or Wisconsin DNR's reporting systems move toward the future. AB - Electronic data transmittal and data warehouses seem like obvious solutions for streamlining reporting systems and managing large bodies of data; however, regulatory agencies like Wisconsin Department of Natural Resources (DNR) face significant barriers in implementation. In addition to the development costs to the Agency, regulators may be limited by the capabilities of the regulated community and the perceived burden for small businesses and communities. Electronic systems can be implemented incrementally if supported by state regulations and processes for insuring data integrity. PMID- 12553078 TI - Data and metadata reporting standards for the U.S. Environmental Protection Agency's PM Supersites Research Program. AB - The EPA Supersites Research Program needs consistency of metadata and data structures to facilitate information sharing among investigators, analysts, and ultimately secondary data users. Under the auspices of NARSTO a successful mechanism was created to develop and implement reporting standards. The development effort included working closely with Supersites data coordinators, investigators, and technical experts, and also leveraging from existing data standards and practices. Overall, the standards are getting good acceptance from the atmospheric research community. PMID- 12553079 TI - Comparison of EPA's QMS to SEI's CMMI. AB - EPA and other government organizations make decisions based on environmental measurements. How good are the data? How well are the data generators performing? What measurements apply to them? How can the data life cycle processes be improved so data generators can continually provide the best data? EPA's Quality Management System requirements go beyond evaluation of environmental data quality itself to examine systems associated with production, collection, processing (validation/verification), transfer, reduction, storage, and retrieval of data throughout a life cycle. This QMS specifies minimum quality requirements for particular environmental programs. But how can you measure and compare programs that go well beyond the minimum, towards optimal quality? This paper compares EPA's requirements for Quality Management Systems (R2) and Project Plans (R5) to the Software Engineering Institute Capability Maturity Model (CMMISM). The CMMISM model provides for growth (staged or continuous) and a comprehensive assessment that is not yet provided in EPA's R2 or R5. Properly implemented, the CMMISM model serves as a quality framework for integrating and aligning organizational processes and implementing a program of continual process improvements. It identifies process areas ("things to do"), and provides measures of performance ("how well things are done") against specific goals and practices. CMMISM uses a Systems Engineering Management approach, built on process models, that helps identify "how good" the system is. Goodness is defined as stages in a complete model for optimal operation. CMMISM provides two methods for evaluating the goodness of the project. The Staged model in CMMISM provides a Maturity Level that is a well-defined evolutionary plateau describing the manner in which a specified set of processes are performed. As the organization advances in maturity, these levels become more defined and processes are tailored for specific project needs. The other method is called the Continuous Model in CMMISM, and it allows you to achieve Capability Levels. These are used to describe how well each project is doing in relationship to the different process areas. There are six Capability Levels from 0-5 that apply to individual process areas. Organizations using the Capability Level approach can select individual process areas that are important to specific projects and work to improve the processes. Improving capability in individual process areas raises the organization's overall quality of products delivered. The Continuous Model, unlike the Staged Model, lets you pick higher maturity level process areas before completing all of the ones below. Environmental measurement programs need to focus on the quality of the systems where data are collected, processed, transferred, and so forth. DynCorp built on the quality foundation from our experience with R2 to successfully implement CMMISM practices in the development of Forms II Lite and other applications. DynCorp is now migrating to the CMMISM model that has evolved from the existing CMM model. The CMMISM model focuses on the full cycle of Requirements Management from identification, development, collection, refinement, analysis, and validation throughout a project life cycle. It also has a more refined focus on the identification, development, collection, analysis, and evaluation of meaningful measurements, so the results can be used to improve a process or product. PMID- 12553080 TI - The importance of a successful Quality Assurance (QA) program from a research manager's perspective. AB - One responsibility of an EPA research manager is to ensure that data from research projects are acquired, processed, and reported in accordance with Quality Assurance (QA) requirements established by the Agency. To meet this responsibility, the research manager should understand Agency QA requirements, require an appropriate, effective Quality Assurance program to ensure that data are of known and acceptable quality for the intended use of the data, and provide support, guidance, and oversight to principal investigators in meeting QA requirements. In addition, the effectiveness of the QA effort can be enhanced if the research manager 1) ensures that principal investigators and other managers are aware that QA is viewed as an essential, integrated component of the research programs; 2) provides adequate resources (people and money) to support an effective Quality Assurance program; 3) encourages cooperative, productive interactions between researchers and Quality Assurance professionals; and 4) maintains oversight so that issues that have the potential for adversely affecting research and Quality Assurance objectives can be negotiated and corrected quickly. This presentation will discuss the Air Pollution Prevention and Control Division's Quality Assurance program and the approaches used to meet Quality Assurance requirements in the Division. The presentation will be a technical manager's perspective of the Division's requirements for and approach to Quality Assurance in its research programs. The presentation will include the design of the QA Team, the roles of members of the QA Team, training and technical aids provided by the QA Team to promote understanding of and adherence to Agency QA requirements, the interactions of the QA Team members with principal investigators, and examples of effective conflict resolution. PMID- 12553081 TI - In search of representativeness: evolving the environmental data quality model. AB - Environmental regulatory policy states a goal of "sound science." The practice of good science is founded on the systematic identification and management of uncertainties; i.e., knowledge gaps that compromise our ability to make accurate predictions. Predicting the consequences of decisions about risk and risk reduction at contaminated sites requires an accurate model of the nature and extent of site contamination, which in turn requires measuring contaminant concentrations in complex environmental matrices. Perfecting analytical tests to perform those measurements has consumed tremendous regulatory attention for the past 20-30 years. Yet, despite great improvements in environmental analytical capability, complaints about inadequate data quality still abound. This paper argues that the first generation data quality model that equated environmental data quality with analytical quality was a useful starting point, but it is insufficient because it is blind to the repercussions of multifaceted issues collectively termed "representativeness." To achieve policy goals of "sound science" in environmental restoration projects, the environmental data quality model must be updated to recognize and manage the uncertainties involved in generating representative data from heterogeneous environmental matrices. PMID- 12553082 TI - Establishing sensitivity requirements for environmental analyses from project data quality objectives. AB - This article proposes a simple strategy for establishing sensitivity requirements (quantitation limits) for environmental chemical analyses when the primary data quality objective is to determine if a contaminant of concern is greater or less than an action level (e.g., an environmental "cleanup goal," regulatory limit, or risk-based decision limit). The approach assumes that the contaminant concentrations are normally distributed with constant variance (i.e., the variance is not significantly dependent upon concentration near the action level). When the total or "field" portion of the measurement uncertainty can be estimated, the relative uncertainty at the laboratory's quantitation limit can be used to determine requirements for analytical sensitivity. If only the laboratory component of the total uncertainty is known, the approach can be used to identify analytical methods or laboratories that will not satisfy objectives for sensitivity (e.g., when selecting methodology during project planning). PMID- 12553083 TI - Transforming an EPA QA/R-2 quality management plan into an ISO 9002 quality management system. AB - The Environmental Protection Agency's (EPA) Office of Emergency and Remedial Response (OERR) requires environmental data of known quality to support Superfund hazardous waste site projects. The Quality Assurance Technical Support (QATS) Program is operated by Shaw Environmental and Infrastructure, Inc. to provide EPA's Analytical Operations Center (AOC) with performance evaluation samples, reference materials, on-site laboratory auditing capabilities, data audits (including electronic media data audits), methods development, and other support services. The new QATS contract awarded in November 2000 required that the QATS Program become ISO 9000 certified. In a first for an EPA contractor, the QATS staff and management successfully transformed EPA's QA/R-2 type Quality Management Plan into a Quality Management System (QMS) that complies with the requirements of the internationally recognized ISO 9002 standard and achieved certification in the United States, Canada, and throughout Europe. The presentation describes how quality system elements of ISO 9002 were implemented on an already existing quality system. The psychological and organizational challenges of the culture change in QATS' day-to-day operations will be discussed for the benefit of other ISO 9000 aspirants. PMID- 12553084 TI - One fish, two fish, we QC fish: controlling data quality among more than 50 organizations over a four-year period. AB - EPA is conducting a National Study of Chemical Residues in Lake Fish Tissue. The study involves five analytical laboratories, multiple sampling teams from each of the 47 participating states, several tribes, all 10 EPA Regions and several EPA program offices, with input from other federal agencies. To fulfill study objectives, state and tribal sampling teams are voluntarily collecting predator and bottom-dwelling fish from approximately 500 randomly selected lakes over a 4 year period. The fish will be analyzed for more than 300 pollutants. The long term nature of the study, combined with the large number of participants, created several QA challenges: (1) controlling variability among sampling activities performed by different sampling teams from more than 50 organizations over a 4 year period; (2) controlling variability in lab processes over a 4-year period; (3) generating results that will meet the primary study objectives for use by OW statisticians; (4) generating results that will meet the undefined needs of more than 50 participating organizations; and (5) devising a system for evaluating and defining data quality and for reporting data quality assessments concurrently with the data to ensure that assessment efforts are streamlined and that assessments are consistent among organizations. This paper describes the QA program employed for the study and presents an interim assessment of the program's effectiveness. PMID- 12553085 TI - Required steps for the validation of a Laboratory Information Management System. AB - The task of managing laboratory data is not a new one. Over the past two decades, the use of Laboratory Information Management Systems (LIMS) has revolutionized how laboratories manage their data. A LIMS is more than software; it has become the workhorse of the laboratory, encompassing laboratory work-flow combined with user input, data collection, instrument integration, data analysis, user notification, and delivery of information and reporting. Types of organizations that utilize LIMS vary greatly from research laboratories to manufacturing laboratories to environmental testing laboratories. Commercially-available LIMS have been around since the 1980s. In addition, many laboratories have designed, implemented, and maintained in-house LIMS. The heart of any LIMS is the software. Like other laboratory systems, the LIMS software is subject to quality control and quality assurance checks. In regulatory environments this associated QA/QC is referred to as "system validation." The primary purpose of system validation is to ensure that the software is performing in a manner for which it was designed. For example, the system acceptance criteria should be established and tested against quantifiable tasks to determine if the desired outcome has been achieved. LIMS features, such as autoreporting, reproducibility, throughput, and accuracy must be quantifiable and verifiable. System validation ensures that the entire system has been properly tested, incorporates required controls, and maintains and will continue to maintain data integrity. Laboratories must establish protocols and standards for the validation process and associated documentation. Although vendors of commercial LIMS perform initial internal system validations, the system must be revalidated whenever the end user, vendor or third party adds modifications or customizations to the LIMS. Currently, detailed guidance regarding system validation of LIMS is not available to the user. The issue is addressed in Good Automated Laboratory Practices (GALP) and National Environmental Laboratory Accreditation Conference (NELAC) documents which indicate specific requirements or recommendations for operational checks and periodic testing; however, it is up to the laboratory to determine suitable methods to accomplish these tasks. Proper validation of a LIMS will allow a laboratory to comply with regulations and also provide comprehensive documentation on the system that is necessary to troubleshoot future problems. PMID- 12553086 TI - Improved quality data systems through the use of standard electronic data deliverables (EDDs) and environmental data assessment software. AB - One of the challenges facing professionals in the environmental arena today is the collection and assessment of large amounts of environmental analytical data. The assessment of the quality of that data is essential as multi-million dollar decisions for environmental site cleanups and/or long term monitoring efforts are made based on the analytical results. Also critical to environmental programs is the sharing and access of data across multiple data users. The ability to share data allows for better use of the limited resources available to clean up and monitor contaminated environmental sites. Standardization of electronic deliverables allows for collection of data from multiple data collectors into a single database for use by numerous data users and stakeholders on a project. This paper discusses the benefits of using a standard EDD deliverable format and use of environmental data assessment software tools to do project planning and data assessment throughout the duration of the environmental project. PMID- 12553087 TI - Quantifying uncertainty: calculating interval estimates using quality control results. AB - EPA's Great Lakes National Program Office (GLNPO) is leading one of the most extensive studies of a lake ecosystem ever undertaken. The Lake Michigan Mass Balance Study (LMMB Study) is a coordinated effort among state, federal, and academic scientists to monitor tributary and atmospheric pollutant loads, develop source inventories of toxic substances, and evaluate the fate and effects of these pollutants in Lake Michigan. A key objective of the LMMB Study is to construct a mass balance model for several important contaminants in the environment: PCBs, atrazine, mercury, and trans-nonachlor. The mathematical mass balance models will provide a state-of-the-art tool for evaluating management scenarios and options for control of toxics in Lake Michigan. At the outset of the LMMB Study, managers recognized that the data gathered and the model developed from the study would be used extensively by data users responsible for making environmental, economic, and policy decisions. Environmental measurements are never true values and always contain some level of uncertainty. Decision makers, therefore, must recognize and be sufficiently comfortable with the uncertainty associated with data on which their decisions are based. The quality of data gathered in the LMMB was defined, controlled, and assessed through a variety of quality assurance (QA) activities, including QA program planning, development of QA project plans, implementation of a QA workgroup, training, data verification, and implementation of a standardized data reporting format. As part of this QA program, GLNPO has been developing quantitative assessments that define data quality at the data set level. GLNPO also is developing approaches to derive estimated concentration ranges (interval estimates) for specific field sample results (single study results) based on uncertainty. The interval estimates must be used with consideration to their derivation and the types of variability that are and are not included in the interval. PMID- 12553088 TI - [Current microbiologic strategy in endocarditis]. PMID- 12553089 TI - [Complications after cardiovascular surgery in patients with diabetes]. PMID- 12553090 TI - [Percutaneous coronary intervention in patients with diabetes mellitus]. AB - The incidence of restenosis after percutaneous coronary intervention (PCI) in patients with ischemic heart disease and diabetes is increased compared to non diabetics. The use of stents has reduced the incidence of restenosis significantly but the effect is not convincing, as restenosis in diabetics predominantly consists of intimal hyperplasia. Even with the use of stents, diabetics have increased morbidity and mortality compared to non-diabetics. The precise mechanism is unknown but may include the metabolic abnormalities characteristic of diabetes. No randomised prospective studies in diabetics compare coronary artery by-pass grafting (CABG) with optimal angioplastic therapy. We await such studies to clarify the conditions for diabetics. Retrospective subgroup analyses indicate a more favourable outcome in diabetics revascularised by CABG. Thus CABG is presumably the optimal revascularisation treatment for diabetics with three-vessel disease. PMID- 12553091 TI - [Molecular modifications of troponin I and T detected in serum from patients with acute myocardial infarction]. AB - INTRODUCTION: Cardiac troponin I and T (cTnI and cTnT) are specific biochemical serum markers for acute myocardial infarction (AMI). However, cTnI diagnostic assays are plagued by difficulties, resulting in > 20-fold differences in measured values. These discrepancies may result from the release of the numerous cTnI modification products that are present in ischaemic myocardium. The resolution of these discrepancies requires an investigation of the exact forms of the troponins present in the bloodstream of patients after myocardial injury. MATERIAL AND METHODS: A Westernblot direct serum analysis protocol was developed that allowed us to detect intact cTnI and a spectrum of up to 11 modified products in the serum from patients with AMI. RESULTS: We document both a cTnI degradation pattern and the existence of phosphorylated cTnI in serum. The number and extent of these modifications reflect patterns similar to the time profiles of the routine clinical serum markers of total creatine kinase, creatine kinase MB, and cTnI (determined by ELISA). Data from in vitro experiments, which were undertaken to study the degradation of human recombinant cTnI and cTnT when spiked in serum, indicate that some modification products present in patient serum existed in the myocardium. DISCUSSION: This pilot study defines, for the first time, what forms of cTnI and cTnT appear in the bloodstream of AMI patients, and it clarifies the lack of standardization between different cTnI diagnostic assays. PMID- 12553092 TI - [Increased use of lipid-lowering agents after implementation of clinical guidelines in general practice]. AB - INTRODUCTION: The preventive efforts with respect to ischaemic heart disease are inadequate, and particularly an under-use of lipid-lowering drugs (LLDs) has been demonstrated. The aim of this study was to examine if the prescribing of LLDs increased after an intervention. The intervention consisted of clinical guidelines on prevention of ischaemic heart disease sent to all general practitioners nationwide, accompanied by a change of reimbursement rules for LLDs. MATERIAL AND METHODS: The clinical guidelines were made by the Danish College of General Practitioners and sent out simultaneously with the introduction of less restrictive reimbursement rules for LLDs in December 1998. Prescriptions for lipid-lowering drugs were analysed based on prescription data in the Odense University Pharmacoepidemiological Database. RESULTS: The consumption of LLDs increased significantly after the intervention. The incidence rate ratio (before/after) was 1.43 (1.35-1.52). Prior to the intervention there was an average annual increase in the prevalence of LLD users of 0.2%. After the intervention the prevalence increased by 0.4%. For patients with diabetes, the figures were 1.2% and 2.3% and for patients with ischaemic heart disease 5.0% and 8.1%, respectively. DISCUSSION: Clinical guidelines on prevention of ischaemic heart disease accompanied by a change of drug reimbursement rules had a significant influence on the prescribing of LLDs. The number of persons treated with LLDs is, however, still too low. PMID- 12553093 TI - [Potential number of rehabilitated cancer patients in Denmark--an estimate]. AB - INTRODUCTION: Over the last 50 years there has been an increase in the number of cancer patients. The aim of this study was to estimate the potential requirements of these patients with regard to rehabilitation. MATERIAL AND METHODS: Patients with non-melanoma skin tumours were excluded from the number of cancer cases reported to the cancer register in 1995. This left a total of 23,124 cases. The patients were grouped according to the following parameters; age, diagnosis, spread, and the degree of radicality of the primary treatment. They were followed up for a period of up to two years. The time span was divided into six month intervals, which were used to calculate the survival rate. Those with a survival rate of less than six months were assumed primarily to need palliation. RESULT: The maximum number of cancer patients needing rehabilitation was estimated to be 70 per cent of the newly diagnosed cases, equivalent to 16,297 patients requiring education, and 5,500 were younger adults with a probable need for work-related rehabilitation. The largest diagnosis groups among the rehabilitation patients are expected to be colorectal cancer, prostate cancer, and lung cancer in males, and breast cancer, coloractal cancer, and gynaecological cancer in females. DISCUSSION: Examination of the literature has shown a substantial need to define and provide evidence-based programmes for cancer rehabilitation. Comprehensive research and development efforts are needed to describe the actual need and the possibility of achieving an effect with these programmes. PMID- 12553094 TI - [Treatment of breast abscess. An analysis of patient material and implementation of recommendations]. AB - INTRODUCTION: Several options for treatment of abscesses in the breast have appeared during the last two decades. No randomised studies comparing these options have been performed. MATERIAL AND METHODS: A retrospective study of the treatment of breast abscesses over a four-year-period. Incision or puncture was employed at the discretion of the responsible doctor. RESULTS: A total of 87 women with various types of breast abscesses were treated by means of incision (42 women equal to 48%) or aspiration of pus by puncture (45 women equal to 52%). After incision which was done in general anaesthesia in 38 women (90%) the patients were discharged after 2.9 days (mean), median 2.0 days, and 39 of 45 women treated by means of puncture were discharged after 2.1 days (mean), median 1.0 day, p = 0.001. In six women, puncture treatment was possible as outpatient procedure. Concerning puncture the majority of patients were treated in local analgesia or without any medication. Recurrent abscess formation was observed in eight women (19%) after incision and in nine women (20%) after puncture. DISCUSSION: The two treatment schedules are effective, but abscess treatment by means of puncture is preferred as the most convenient method for the patient and as a cost-effective treatment. Finally, recommendations for the future schedule of breast absess treatment are specified. PMID- 12553095 TI - [Thyroidectomy. A prospective assessment of hemithyroidectomy]. AB - AIM: The results of hemithyroidectomy and total thyroidectomy were assessed in a prospective study. Total thyroidectomy was compared to hemithyroidectomy and contralateral resection. The rate of vocal cord palsy, hypocalcaemia, changes in PTH, and the frequency of sequelae were used to monitor the results. MATERIAL AND METHODS: One hundred and twenty patients underwent hemithyroidectomy for unilateral thyroid lesions and 80 hemithyroidectomy contralateral resection (35 patients) or total thyroidectomy (45 patients) for bilateral disease. Plasma calcium and PTH were measured pre- and post-operatively, and the need for calcium and vitamin D supplementation was registered. RESULTS: No permanent vocal cord palsy was encountered. Calcium substitution was not needed after hemithyroidectomy but for more than one year in 9% after hemithyroidectomy and contralateral resection, and in 7% after total thyroidectomy (NS). One needed substitution 2 years after total thyroidectomy. Pre- and postoperative plasma-PTH was found unchanged after hemithyroidectomy and after total thyroidectomy. DISCUSSION: Hemithyroidectomy and total thyroidectomy are safe procedures with few side effects, if a meticulous dissection is performed. Hypocalcaemia following bilateral operations is usually transient and the need for calcium and vitamin D supplementation is low and usually related to the underlying disease rather than to the operation. Total thyroidectomy will remove the target organ for the immune response in patients with hyperthyroidism and the risk of toxic ophthalmopathy is minimised. Lastly, the risk of recurrent disease is eliminated. Six per cent had minor and probably transient local complaints at control three months after the operation. PMID- 12553096 TI - [Symptoms resembling acute myocardial infarction caused by tracheal compression because of enlarged thyroid gland containing metastasis from pancreatic cancer]. AB - This case describes a 78-year-old man, suffering from cancer of the prostate. The patient had had a thyroid resection 22 years earlier. He was admitted with symptoms of myocardial infarction caused by tracheal compression by an enlarged thyroid. After tracheostomy the patient recovered promptly. Resection of the thyroid revealed a little area of adenocarcinoma cells representing a metastasis from the prostate together with a Hurthle cell tumour. PMID- 12553097 TI - [Pericardial cyst--a rare cause of thoracic pain]. AB - A case of a 10 cm in diameter and 450 ml large pericardial coelom cyst causing chest pain in a previously healthy 38-year-old man is reported. The cyst was diagnosed by transthoracic echocardiography and computerized tomography. The patient recovered without complications following surgical removal of the cyst. PMID- 12553098 TI - [Radical prostatectomy versus observation for localized prostatic cancer]. PMID- 12553099 TI - [Surgery crisis--seen from a fundamental point of view]. PMID- 12553100 TI - [The shortage of surgeons is self-inflicted!]. PMID- 12553101 TI - [It smells!]. PMID- 12553102 TI - [Adverse effects of vitamin D3]. PMID- 12553103 TI - [The search for yet unrecognized infectious pathogens]. PMID- 12553104 TI - [Prion diseases]. PMID- 12553105 TI - [Epstein-Barr virus and disseminated sclerosis: causative or secondary associations?]. PMID- 12553106 TI - [Guillain-Barre syndrome and Campylobacter]. PMID- 12553107 TI - [Epstein-Barr virus and nasopharyngeal carcinoma]. PMID- 12553108 TI - [Autoimmune thyroiditis--an infectious disease?]. AB - The aim was to review existing evidence of a possible role of infectious agents in the pathogenesis of autoimmune thyroid disease. Autoimmune thyroid disease is a polygenic, multifactorial disease in which genetically susceptible individuals are exposed to an environmental insult resulting in immune system activation. Different viruses (influenza B, rubella, retrovirus) have been associated with thyroiditis, but no single agent appears to be causative. There is no firm evidence of infection being an important trigger of autoimmune thyroid disease, and it has not been possible to isolate a microorganism neither by culture nor by molecular identification. Infection may be a precipitating factor in the development of autoimmune thyroid disease. PMID- 12553109 TI - [Reduction of the expansion rate of small abdominal aortic aneurysms with roxithromycin. Results from a randomized controlled trial]. AB - INTRODUCTION: Macrolide treatment has been reported to reduce the risk of recurrent ischaemic heart disease. The influence of a macrolide on the expansion rate of small abdominal aortic aneurysms (AAA) is unknown at present. The aim of this study was to investigate the effect of roxithromycin on the expansion rate of small AAA. MATERIALS AND METHODS: A total of 92 patients with a small AAA were recruited from two populations. One population consisted of 6.339 men aged 65-73 years who were offered participation in a mass screening programme for AAA at the local hospital. From this population 66 subjects were recruited. The remaining 26 were recruited from among 49 subjects diagnosed at interval screening for an initial aortic diameter between 25 mm and 29 mm. The patients were randomized to receive either oral roxithromycin 300 mg once daily for 28 days or matching placebo, and followed for a mean of 1.5 years. RESULTS: During the first year the mean annual expansion rate of AAA was reduced by 44% in the macrolid group (1.56 mm/year) compared to 2.80 mm/year after placebo (p = 0.02). During the second year the difference was only 5%. Multiple linear regression analysis showed that roxithromycin treatment and initial AAA size were significantly related to AAA expansion when adjusted for smoking, diastolic blood pressure, and IgA level > or = 20. The logistic regression analysis confirmed a significant difference in expansion rates above 2 mm annually between the intervention and placebo groups, OR = 0.09 (95% CI: 0.01-0.83). DISCUSSION: In comparison to placebo, roxithromycin 300 mg daily for four weeks reduced the expansion rate of AAA. PMID- 12553110 TI - [Atherosclerosis and Chlamydia pneumoniae]. AB - Chlamydia pneumoniae could be associated with the risk of developing atherosclerosis and an increased risk of thromboembolic complications. However, the evidence of an association seems to be declining and there is no evidence of causality. The effect of antibiotic treatment in cardiovascular disease has been explored in epidemiologic studies and in randomised controlled trials. Data suggest a protective but short-lasting effect of macrolide antibiotics on cardiovascular disease. The effect could be the result of anti-bacterial as well as anti-inflammatory properties. Ongoing larger and longer lasting treatment trials could provide better measures of the effects of antibiotic treatment, although they will not clarify the role of C. pneumoniae. Currently, there is no indication for treating cardiovascular disease with antibiotics. PMID- 12553111 TI - [Epstein-Barr virus and Hodgkin's lymphoma]. AB - The suspicion that Hodgkin's lymphoma, previously known as Hodgkin's disease, may have an infectious etiology has existed for many years. The assumption rests on epidemiological characteristics, in particular a strong correlation between socio economic status and risk of Hodgkin's lymphoma in young adults, as well as both serological and molecular biological evidence that the Epstein-Barr virus is involved in the development of up to 50% of all cases of Hodgkin's lymphoma. We present the current epidemiological, serological and molecular biological evidence for an infectious etiology to Hodgkin's lymphoma with emphasis on the association with Epstein-Barr virus. PMID- 12553112 TI - [Infectious causes of non-Hodgkin lymphomas]. AB - The etiology to non-Hodgkin's lymphoma remains incompletely understood. Chronic infection with certain viruses and bacteria has attracted interest in recent years because of the association with lymphoma development. In this article we present an overview of the current evidence of infectious causes to non-Hodgkin's lymphomas. PMID- 12553113 TI - [Herpesvirus 8, Kaposi sarcoma and other lymphoproliferative diseases]. PMID- 12553114 TI - [Kawasaki syndrome--an infectious disease?]. PMID- 12553115 TI - [Type 1 diabetes mellitus--an infectious disease?]. AB - Rubella infection in utero is associated with a high risk of later development of type 1 diabetes. The mechanism is still unclear although the balance between the so-called Th1 and Th2 immune response seems to be important. Whether viral infections are involved in the pathogenesis of the common type of immune mediated type 1 diabetes is still unclear. Prospective serological investigations suggest a higher frequency of enteroviral infections in utero or in early childhood in children later developing type 1 diabetes. There is a need for confirmatory investigations of these observations especially with regard to demonstration of viral genome. PMID- 12553116 TI - [Hepatitis B--a viral oncogene?]. AB - Infection with hepatitis B virus may develop into a chronic carrier state, which may result in chronic hepatitis, cirrhosis and hepatocellular carcinoma, but the exact mechanism of the development of hepatocellular carcinoma is still not known. The question is whether it is hepatitis B virus itself or the cellular changes due to persistent hepatitis B virus infection that cause malignant changes? The purpose of this article was to describe the natural history of hepatitis B virus infection together with epidemiological, serological, and molecular data that show a connection between chronic hepatitis B virus infection and the development of hepatocellular carcinoma. PMID- 12553117 TI - [Peptic ulcer and stomach cancer--infectious diseases caused by Helicobacter pylori?]. PMID- 12553118 TI - [Inflammatory bowel disease--do microorganisms play a role?]. AB - This review focuses on the potential pathogenic role of microorganisms in relation to inflammatory bowel diseases, i.e. Crohn's disease and ulcerative colitis. Pathogenic microorganism such as Mycobacterium paratuberculosis, measles and mumps viruses, Epstein-Barr virus, and Listeria monocytogenes are discussed, as well as involvement of the normal intestinal flora. Furthermore, the influence of microorganisms in experimental animal colitis models is discussed. The available results are inconclusive, but there seems to be basis for proposing the hypothesis that the inflammation in inflammatory bowel disease reflects an immune imbalance with loss of tolerance for normally harmless antigens in the mucosal microflora. PMID- 12553119 TI - [Human papillomavirus and the risk of anogenital cancer]. AB - Strong evidence suggests that virtually all cases of cervical cancer are caused by infection with oncognenic types of human papillomavirus (HPV). HPV is also found frequently in neighbouring anogenital epithelia and their neoplastic lesions. This review describes the general aspects of the natural history of anogenital HPV infections and summarises the evidence for an association between HPV and cancers of the uterine cervix, vulva, anus and penis. PMID- 12553120 TI - [Pseudomembranous upper respiratory tract infection caused by herpes simplex virus in an adult]. AB - We report a case of viral laryngitis due to HSV in an adult. The patient was a 61 year-old female alcoholic. She presented with fever and stridor and developed sepsis and multiorgan failure. She was treated with antibiotics to no effect and died 14 days after hospitalisation. Post mortem histology of the larynx showed an acute pseudomembranous inflammation with the classical morphology of HSV infection. An immunohistochemical reaction for HSV was positive. PMID- 12553121 TI - [A case of pericarditis in a patient with brucellosis]. AB - A case of brucellosis pericarditis is described in a patient presenting with undulant fever, arthralgias, myalgias, weight loss, and anemia. Two cultures of blood showed growth of Brucella abortus. The patient had no retrosternal pain, but stethoscopy of the heart revealed a murmur and pericarditis was echocardiographically diagnosed. PMID- 12553122 TI - [Treatment with testosterone]. PMID- 12553123 TI - [Placebo effect: arthroscopic lavage and debridement of knee osteoarthritis]. PMID- 12553124 TI - [Cauliflower ears--a comment]. PMID- 12553125 TI - The psychiatrist and the clinical practice of psychiatry in an uncertain environment: looking ahead. PMID- 12553126 TI - Taking aim at posttraumatic stress disorder: understanding its nature and shooting down myths. PMID- 12553127 TI - Epidemiologic studies of trauma, posttraumatic stress disorder, and other psychiatric disorders. AB - This paper reviews recent epidemiologic studies of posttraumatic stress disorder (PTSD) in the general population. Estimates of the prevalence of exposure to traumatic events vary with the method used to ascertain trauma exposure and the definition of the stressor criterion. Changes in the DSM-IV definition of "stressor" have increased the number of traumatic events experienced in the community that can be used to diagnose PTSD and thus, the number of PTSD cases. Risk factors for PTSD in adults vary across studies. The 3 factors identified as having relatively uniform effects are 1) preexisting psychiatric disorders, 2) a family history of disorders, and 3) childhood trauma. In civilian populations, women are at a higher risk for PTSD than are men, following exposure to traumatic events. Most community residents have experienced 1 or more PTSD-level traumas in their lifetime, but only a few succumb to PTSD. Trauma victims who do not succumb to PTSD are not at an elevated risk for the subsequent onset of major depression or substance use disorders, compared with unexposed persons. PMID- 12553128 TI - PTSD and the experience of pain: research and clinical implications of shared vulnerability and mutual maintenance models. AB - It is common for individuals with symptoms of posttraumatic stress disorder (PTSD) to present with co-occurring pain problems, and vice versa. However, the relation between these conditions often goes unrecognized in clinical settings. In this paper, we describe potential relations between PTSD and chronic pain and their implications for assessment and treatment. To accomplish this, we discuss phenomenological similarities of these conditions, the prevalence of chronic pain in patients with PTSD, and the prevalence of PTSD in patients with chronic pain. We also present several possible explanations for the co-occurrence of these disorders, based primarily on the notions of shared vulnerability and mutual maintenance. The paper concludes with an overview of future research directions, as well as practical recommendations for assessing and treating patients who present with co-occurring PTSD or chronic pain symptoms. PMID- 12553129 TI - Electroconvulsive therapy training in Canada: a call for greater regulation. AB - OBJECTIVE: A national survey of senior psychiatric residents' experience of electroconvulsive therapy (ECT) training was undertaken in Canada in 1989. Since then, guidelines have been published in Canada and the US with respect to ECT training in psychiatry residency programs. This study examines whether there has been any change in the Canadian resident ECT training experience through a second national survey. METHODS: Confidential questionnaires assessing the training in and the attitudes toward ECT were sent to all 133 psychiatric residents-in their final year of training in Canadian medical schools. RESULTS: We attained a response rate of 68.3%. Of respondents, 88% felt that theoretical and practical ECT training should be a mandatory component of psychiatric residency programs. No marked improvements have occurred with respect to didactic and bedside training in ECT, compared with the original 1989 study. Only 18% of respondents reported that they feel completely competent administering ECT. Despite this, 59.3% anticipate working in an inpatient setting. CONCLUSIONS: The publication of training guidelines has made little impact on training in and attitudes toward ECT in senior psychiatric residents in Canada. Despite senior residents' acknowledgement of ECT's value, few feel competent to administer ECT after completing their training. PMID- 12553130 TI - Interrater reliability of the Fitness Interview Test across 4 professional groups. AB - OBJECTIVE: This study investigated the interrater reliability of the Fitness Interview Test (FIT), revised edition, a semistructured interview that assesses fitness to stand trial. METHOD: Physicians, forensic psychologists, nurses, and graduate students in psychology were trained in the FIT, and they subsequently viewed 2 videotaped interviews of actual fitness assessments. Using the FIT, they rated the fitness of each defendant portrayed in the videotapes. RESULTS: For overall judgment of fitness, the average intraclass correlation based on the full samples of raters was found to be 0.98, and for most items on the FIT, intraclass correlations fell within the 0.80 s and 0.90 s. Reliability estimates were high across professional groups. CONCLUSIONS: Overall, this study provides further support for the psychometric properties of the FIT, as well as for the ability of various professionals to conduct reliable fitness assessments using the FIT. PMID- 12553131 TI - Posttraumatic symptoms and disability in paramedics. AB - OBJECTIVE: The concern that secondary gain may result in an overreporting of trauma symptoms in those seeking compensation or taking stress leave from work has raised questions about the relation between posttraumatic stress and disability. This study attempts to examines the relation between traumatic stress symptoms and the use of work leave in an anonymous sample of emergency-service workers who are not currently seeking compensation. METHOD: A total of 86 paramedics completed questionnaires that addressed exposure to traumatic events, use of mental health stress leave, social support, current level of distress, and personality patterns. Comparisons were made between groups who had used mental health stress (MHS) leave and those who had not. Logistic regression was used to determine the best predictors of using leaves. RESULTS: Current levels of social support were associated with previous use of mental health stress leave. In addition, significantly more individuals who had taken MHS leave in the past reported posttraumatic stress symptoms in the high or severe range. People with personality patterns characterized by suspiciousness, hostility, and isolation and having a tendency toward demanding, controlling, and manipulative behaviour in relationships were also more likely to have taken an MHS leave. CONCLUSION: Although social support and trauma symptoms were associated with the use of MHS leave, in this study, personality style was the strongest factor differentiating those individuals who took MHS leave from those who did not. PMID- 12553132 TI - Antipsychotic medication during pregnancy and lactation in women with schizophrenia: evaluating the risk. AB - OBJECTIVE: To review studies investigating the following: whether exposing developing infants to antipsychotic medication during pregnancy and lactation is associated with increased risks of teratogenic, neonatal, and long-term neurobehavioural sequelae; whether schizophrenia itself affects pregnancy outcome; and whether the course of schizophrenia symptoms is altered by pregnancy and lactation. METHOD: We summarize the results from articles identified via a MedLine search for the period January 1, 1966, to December 1, 2001. RESULTS: Women with schizophrenia are at increased risk for poor obstetrical outcomes, including preterm delivery, low birth weight, and neonates who are small for their gestational age. A lack of information in the literature makes it difficult to comment on the relative risk of exposing developing infants to atypical antipsychotics. However, typical antipsychotics appear to carry an increased risk of congenital malformations when the fetus is exposed to phenothiazines during weeks 4 to 10 of gestation. Lack of information also precludes an understanding of whether changes associated with pregnancy and lactation significantly alter the course of schizophrenia symptoms. CONCLUSION: Research is needed so that physicians may more accurately inform women about the relative risks of using antipsychotic medications during pregnancy and lactation. Increased knowledge about the risks of medication exposure will allow clinicians to limit treatment to situations in which the risk of untreated maternal illness outweighs the risk of exposing a developing infant to medications. PMID- 12553133 TI - Antidepressants and the risk of breast cancer. AB - BACKGROUND: A recent national newspaper article highlighted 2 published research papers that suggest an association between antidepressants and an increased risk of breast cancer. The authors of the 2 papers recommend switching or avoiding the antidepressants implicated in their studies. METHOD: We critically review these papers and, based on our review, consider what clinical practices, if any, should be modified as a result of their findings. RESULTS: Both articles are based upon case-control studies. In the first paper, the authors examine the relation between tricyclic antidepressants (TCAs) and an increased risk of breast cancer. The study upon which the paper is based has several design strengths, and the paper presents findings that have biological plausibility. However, the conclusions are weakened by the lack of accounting for potential confounding factors and multiple statistical comparisons. In the second paper, the authors combined survey and administrative data to examine the association between antidepressant use and breast cancer risk. The press article notwithstanding, the second paper does not find a significant association between specific antidepressants and an increased risk of breast cancer, after adjusting for potential confounders. There are significant methodological limitations to the research upon which the paper is based. CONCLUSIONS: The finding of an association between TCA use and increased risk of breast cancer merits further testing using stronger research designs. However, because of the methodological concerns described, the 2 papers we review provide insufficient evidence to guide practitioners to change clinical practice. PMID- 12553134 TI - An analysis of religion and mental illness. PMID- 12553135 TI - Re: Canadian psychiatric inpatient religious commitment: an association with mental health. PMID- 12553136 TI - Oxcarbazepine treatment of posttraumatic stress disorder. PMID- 12553137 TI - Voice mail as a transitional object in the treatment of borderline personality disorder. PMID- 12553138 TI - Critical appraisal of extended treatment studies in attention-deficit hyperactivity disorder. PMID- 12553139 TI - Gabapentin-induced paradoxical exacerbation of psychosis in a patient with schizophrenia. PMID- 12553140 TI - Probable dementia with Lewy bodies and risperidone-induced delirium. PMID- 12553141 TI - Re: Schizophrenia, suicide, and blood count during treatment with clozapine. PMID- 12553142 TI - Re: Bilsbury and others. More on the phenomenology of perfectionism- incompleteness. PMID- 12553143 TI - [Ritual circumcision]. PMID- 12553144 TI - [Surgery performed on healthy genitals of children--for how long? Debate about the conflict between parental rights and basic human rights of children]. PMID- 12553145 TI - [The bear is sleeping.... Adaptation connected to hibernation]. PMID- 12553146 TI - [The time of pigs. Genes of pigs are to be surveyed in an extensive Danish Chinese cooperation]. PMID- 12553147 TI - [Physicians in Australia. Sexual life is suffering or experience from a private hospital system]. PMID- 12553149 TI - [I will see it because I believe it!]. PMID- 12553148 TI - [I think I that I see]. PMID- 12553150 TI - [Tobacco and murder--the first case of nicotine poisoning proved in a homicide]. PMID- 12553151 TI - [Is there an indication for the treatment of biochemically euthyroid, but clinically hypothyroid individuals with thyroid hormone?]. PMID- 12553152 TI - [Doctors' club 1]. PMID- 12553154 TI - [Unnecessary words in dictated epicrises]. PMID- 12553153 TI - [Literature search on nocturia]. PMID- 12553155 TI - [Tick-off-formularies concerning single subsidy of certain drugs--with implemented criteria]. PMID- 12553156 TI - [Orthotopic liver transplant in rats. Surgical technique, complications and treatment]. AB - The orthotopic rat liver transplant model is a widely used technique in transplantation research. It has many advantages over other animal transplant models because of its availability and low cost. However, it must be emphasized that success with the rat model requires thorough training. The aim of this paper is to describe the microsurgical technique involved in 60 rat liver transplants and to discuss the complications and their treatments. Forty-nine liver transplants were performed at the Experimental Laboratory of the University Hospital, Ontario, Canada (ELUH) and 11 were performed at the Laboratorio de Trasplante de Organos de la Facultad de Ciencias Medicas de La Plata, Buenos Aires. Argentina (LTO). Among the transplants performed at the ELUH, the observed complications were haemorrhage (n = 4), pneumothorax (n = 1), anastomotic failure (n = 15), bile leak (n = 3), and bile duct necrosis (n = 9). The remaining 17 rats at the ELUH were healthy at day 7 after surgery. Animal survival immediately postop, at 24 hours postop and at 7 days postop was achieved with the 9th, 20th and 21st transplants respectively. At the LTO, 3 rats died as a result of anaesthetic complications. Seven-day animal survival was achieved with the 11th transplant. We beleive that the description of the orthotopic rat liver transplantation technique, as well as the discussion regarding complications and their management, can be useful for researchers interested in performing liver transplantation in rats. PMID- 12553157 TI - [Acute experimental pancreatitis in the opossum. The role of early splenectomy in the inflammatory response]]. AB - BACKGROUND: The role of the Autonomous Nervous System in the immunologic and inflammatory response is still an issue of discussion. Furthermore, the physiopathologic mechanisms involved are still unknown. Acute pancreatitis (AP) does not escape this disconcert. In fact, like in every severe acute inflammatory process, its discontrol could be responsible of the high morbidity and mortality rates. OBJECTIVE: To assess to which degree bilateral splanchnicectomy changes the course of acute inflammatory response in AP. METHOD: Prospective research. RESULTS: The following parameters were evaluated: red blood cell count, white blood cell count, calcium, glucemia, urea, aminase, lypase and liver enzymes. Macroscopy and microscopy views of the pancreas were also obtained. The leucocitary response was abolished, and the calcium levels dropped to a lesser degree. CONCLUSIONS: Bilateral splanchnicectomy prior to unchaining AP had a beneficial effect, Its mechanism of action could have been through the disconnection of the respective reflex arches. PMID- 12553158 TI - [Paracetamol overdose and experimental cholestasis induced gene expression regulation that codified P-Glycoprotein type 1 in rats]. AB - Excretion of lipophilic cationic toxic compounds from the interior of the hepatocyte to the bile is mediated by P-Glycoprotein. It is an integral protein located in the bile canaliculi. The present work study the hepatic expression of P-Glycoprotein in different models of experimental liver disease: Acute paracetamol intoxication, extrahepatic cholestasis and cholestasis followed by acute paracetamol intoxication. mRNA was isolated from liver tissue, and the expression of Pg-p was assessed by Northern blot. It is concluded that the hepatic expression of mdr1b is increased in the experimental liver diseases when compared to controls. On the other hand, mdr2 expression was similar between the different groups. PMID- 12553159 TI - [A new method for detection and erradication of Helicobacter pylori infection by stool antigens test]. AB - Nowadays technics for Helicobacter pylori detection in stools like culture, and PCR, are expensive and difficult to perform. The aim of this study was to evaluate ELISA test efficacy for detection of H. Pylori antigens in stools comparing this results with standarized technics like histology (Giemsa), ureasa test and UBT C 14. 26 patients were evaluated in this study, ages between 15-75 with upper gastrointestinal symptoms; all of them required gastroduodenal endoscopy, status H. Pylori was determined with methods upon mentioned. 24 hours after endoscopy H. Pylori antigens in stools with the technique Premier Platinum Htsa, Elisa were determined. The detection of H. Pylori antigens in stools accurately identified active H. Pylori infection. The performance characteristics of this non-invasive method was similar in sensibility and specificity to conventional tests. PMID- 12553160 TI - [Sexual transmission of Hepatitis C virus]. AB - Fifty heterosexual partners out of 50 patients (index cases) without confirmed or probable risk factors of parenteral transmission by HCV were studied from 1/1997 to 1/2001. The index cases were HCV Ab (+) (EIA 3rd Abbott and PCR + by RT-nested PCR). The couples with probable or confirmed risk factors of parenteral transmission were strictly excluded. No case of HCV infection was found, suggesting that sexual via wouldn't be a direct transmission of HCV. PMID- 12553161 TI - [Liver leiomyosarcoma]. AB - Hepatic tumors are infrequent. Sarcomas represent less than 2% of them, and Leiomyosarcoma (LMS) is a certain rarity. The frequency grows from primitive tumors to metastatic tumors and in the last years, in relation with different immunodeficiencies. A case of a patient diagnosed in the postoperative and assisted in two relapses until her death, is reported. The possible aetiologies are analyzed in the hepatic gland that does not have muscle of its own, as well as in diagnostic and therapeutic modalities, insisting in the lack of rules, because of the low number of publications. Morphologic variations between the primitive tumor and its relapses are also shown. PMID- 12553162 TI - Up to date. Gastroesophageal reflux disease (GERD) influence of obesity. AB - In this paper the author reviews the epidemiology of gastroesophageal reflux disease, as well as the etiology, methods of study, esophageal pathology association, and medical and surgical treatment of obesity as related to GERD. PMID- 12553163 TI - [Hepatitis A in childhood. The tip of an infectious disease iceberg]. AB - The objectives of this study were to analyze the seroepidemiologic prevalence of Hepatitis A Virus (HAV) in children of the city of Resistencia by means of specific antibody detection, relate these data with the socio-sanitary conditions, and discuss vaccine strategies. Two hundred and eighty eight children between 2 and 14 years of age, with a mean of 6.6 years, of both sexes and with no patent liver disease were studied. Blood samples were taken, and the presence of total anti-HAV antibodies was determined. A prevalence of 83.3% was found with no significant differences between sexes. When age groups were compared, antibodies were found in 57.3% of children between 2 and 4 years of age, 90.8% in the 5 to 9 group, and 96.6% in the 10 to 14 group. It was seen that the precarious system of excreta elimination, the lack of potable water in the dwellings, and the absence of sanitary devices, were statistically associated with the high prevalence of HAV infection. In view of the high endemicity found in the first years of life, and considering this disease as a marker of other pathologies with a similar pattern of dissemination, these data may represent the tip of an iceberg holding a broad base of accompanying infections with a high impact in the health of the population. A simultaneous approach towards anti HAV vaccination in young children, and the political decision of improving socio sanitary conditions and decreasing poverty indexes, should be promptly implemented. PMID- 12553164 TI - [Iron, deficiency anemia, an approach from the gastroenterology]. PMID- 12553165 TI - Brain iron homeostasis. AB - Iron is essential for virtually all types of cells and organisms. The significance of the iron for brain function is reflected by the presence of receptors for transferrin on brain capillary endothelial cells. The transport of iron into the brain from the circulation is regulated so that the extraction of iron by brain capillary endothelial cells is low in iron-replete conditions and the reverse when the iron need of the brain is high as in conditions with iron deficiency and during development of the brain. Whereas there is good agreement that iron is taken up by means of receptor-mediated uptake of iron-transferrin at the brain barriers, there are contradictory views on how iron is transported further on from the brain barriers and into the brain extracellular space. The prevailing hypothesis for transport of iron across the BBB suggests a mechanism that involves detachment of iron from transferrin within barrier cells followed by recycling of apo-transferrin to blood plasma and release of iron as non transferrin-bound iron into the brain interstitium from where the iron is taken up by neurons and glial cells. Another hypothesis claims that iron-transferrin is transported into the brain by means of transcytosis through the BBB. This thesis deals with the topic "brain iron homeostasis" defined as the attempts to maintain constant concentrations of iron in the brain internal environment via regulation of iron transport through brain barriers, cellular iron uptake by neurons and glia, and export of iron from brain to blood. The first part deals with transport of iron-transferrin complexes from blood to brain either by transport across the brain barriers or by uptake and retrograde axonal transport in motor neurons projecting beyond the blood-brain barrier. The transport of iron and transport into the brain was examined using radiolabeled iron-transferrin. Intravenous injection of [59Fe-125]transferrin led to an almost two-fold higher accumulation of 59Fe than of [125I]transferrin in the brain. Some of the 59Fe was detected in CSF in a fraction less than 30 kDa (III). It was estimated that the iron-binding capacity of transferrin in CSF was exceeded, suggesting that iron is transported into the brain in a quantity that exceeds that of transferrin. Accordingly, it was concluded that the paramount iron transport across the BBB is the result of receptor-mediated endocytosis of iron-containing transferrin by capillary endothelial cells, followed by recycling of transferrin to the blood and transport of non-transferrin-bound iron into the brain. It was found that retrograde axonal transport in a cranial motor nerve is age-dependent, varying from almost negligible in the neonatal brain to high in the adult brain. The principle sources of extracellular transferrin in the brain are hepatocytes, oligodendrocytes, and the choroid plexus. As the passage of liver-derived transferrin into the brain is restricted due to the BBB, other candidates for binding iron in the interstitium should be considered. In vitro studies have revealed secretion of transferrin from the choroid plexus and oligodendrocytes. The second part of the thesis encompasses the circulation of iron in the extracellular fluids of the brain, i.e. the brain interstitial fluid and the CSF. As the latter receives drainage from the interstitial fluid, the CSF of the ventricles can be considered a mixture of these fluids, which may allow for analysis of CSF in matters that relate to the brain interstitial fluid. As the choroid plexus is known to synthesize transferrin, a key question is whether transferrin of the CSF might play a role for iron homeostasis by diffusing from the ventricles and subarachnoid space to the brain interstitium. Intracerebroventricular injection of [59Fe125I]transferrin led to a higher accumulation of 59Fe than of [125I]transferrin in the brain. Except for uptake and axonal transport by certain neurons with access to the ventricular CSF, both iron and transferrin were, however, restricted to areas situated in close proximity to the ventricular and pial surfaces. In particular, transferrin injected into the ventricles was never observed in regions distant from the CSF. It was concluded that choroid plexus-derived transferrin is not likely to play a significant role for binding and transporting iron in the brain interstitium. Transferrin secretion from oligodendrocytes probably plays the key role in this process. In the third part of the thesis, the uptake of iron by neurons devoid of projections beyond the blood-brain barrier and glia is addressed. Given the fact that the demonstration of plasma proteins in brain sections can be hampered by several methodological factors, a mapping of the cellular distribution of transferrin in the brain was performed employing extensive use of tissue processing and staining protocols. In order to aid in the understanding of cellular iron uptake in the intact brain, attempts were made to identify iron, transferrin, and transferrin receptors at the light microscopic level. Consistent with the widespread distribution of transferrin receptors in neurons, the ligand transferrin was also found in neurons throughout the CNS. When examined at high resolution, transferrin was found to be distributed to the cytoplasm of neurons, exhibiting a dotted appearance, which is probably consistent with a distribution in the endosomallysosomal system. In contrast to the consistent presence of transferrin receptors on neurons, it was not possible to detect transferrin receptors on glial cells. Related to these observations, the presence of non transferrin-bound iron in the brain suggests that glial cells may take it up by a mechanism that does not involve the transferrin receptor. The widespread distribution of ferritin in glial cells clearly indicates that the glial cells acquire iron. Dietary iron-overload did not change the distribution of transferrin receptors or ferritin in the brain. By contrast, iron deficiency altered the cellular content of these proteins so that transferrin receptors were higher and ferritin lower. The transport of iron from brain to blood was addressed in the last part of the thesis. It was found that in the case of iron and transferrin, there is no evidence showing other significant routes of transport from the brain extracellular fluid into the blood than drainage to the ventricular system followed by export to the blood via the arachnoid villi. The turnover of transferrin in the CSF was found to be very high. For reasons mentioned above, transferrin of the CSF is of little significance for transport and cellular delivery of iron to transferrin receptor-expressing neurons. Instead, transferrin of the CSF probably plays a significant role for neutralization and export to the blood of metals, including iron. Once appearing in blood, transferrin of the CSF was degraded at the same rate as intravenously injected transferrin, which indicates that the transferrin of CSF is not altered to an extent that changes its catabolism during the passage from CSF to blood plasma. The metabolism of iron in the developing brain was found to differ markedly when compared to that of the adult brain. A developing regulated transfer of iron to the brain was reflected morphologically by a higher content of transferrin receptors and non-heme iron in endothelial cells of the developing rat brain than in the adult. Neurons had a very low level of transferrin receptors. After about 20 days of age, iron transport into the brain decreased rapidly, and transferrin receptors appeared on neurons. Iron and transferrin injected into the ventricular system of the developing brain were much more widely distributed in the brain parenchyma than in the adult brain. This high accumulation of substances injected into the ventricles in young animals is probably due to the lower rate of production and turnover of CSF, which will increase the time available for diffusion of proteins into the brain parenchyma, thus giving neurons of the developing brain the opportunity to take up transferrin originating from the CSF. PMID- 12553166 TI - The effect of surgical stress, endotoxin induced sepsis and extracorporeal circulation on the cell mediated immunity. PMID- 12553167 TI - The molecular genetic basis and diagnosis of familial hypercholesterolemia in Denmark. AB - Normal function of the hepatic low-density lipoprotein (LDL) receptor is obligate for normal levels of plasma LDL cholesterol. The LDL receptor regulates the concentration of plasma LDL cholesterol by internalizing apolipoprotein B-100- and apolipoprotein E-containing lipoproteins by receptor-mediated endocytosis. Mutations in the gene encoding the LDL receptor protein give rise to one of the most common classical autosomal dominant inherited disorders in man, familial hypercholesterolemia (FH). The estimated prevalence of heterozygous FH is 0.2% (1:500) in most populations of the world including the Danish. Worldwide, an estimated ten million people are afflicted with FH and in Denmark there are approximately 10,000 subjects with heterozygous FH. Persons with heterozygous FH are characterized by a severely elevated concentration of LDL cholesterol in plasma starting in early childhood, tendon xanthomas and a markedly increased risk of premature coronary heart disease (CHD). Adequate control of plasma LDL cholesterol levels can be achieved in most patients with heterozygous FH, and to a lesser extent in the very rare cases with homozygous FH, using combinations of diet, drug therapy and selective LDL-apheresis. So, it is very important that physicians be aware of this relatively common disorder since there is good evidence that early diagnosis and cholesterol-lowering therapy will delay or even prevent CHD in persons with FH. A large majority of these persons, however, are still not diagnosed or adequately treated. It is believed that the diagnostic abilities molecular biology has to offer will provide the impetus for correcting this situation. The aims of the studies behind the present thesis, therefore, were to obtain important knowledge about current mutation detection technology, prevalence and spectrum of LDL receptor gene mutations in Denmark, methods to evaluate pathogenicity of LDL receptor gene mutations, relationship between FH genotype-phenotype, and clinical versus DNA diagnosis in the Danish FH population. Among different relative laborious and expensive scanning methods for unknown gene mutations we have shown that the polymerase chain reaction (PCR) single-strand conformation polymorphism (SSCP) analysis is a highly efficient and sensitive technique for detection of mutations in the 18 exons including intronic splice-site sequences and the promoter region of the LDL receptor gene, reserving DNA sequencing to the exons revealing variant SSCP patterns. Southern blot analysis or long distance PCR analysis are necessary to identify large gene re arrangements in the LDL receptor gene in FH patients in whom SSCP analysis did not reveal any smaller sequence alterations. Worldwide, about 700 different mutations in the LDL receptor gene have been reported and in the Danish FH population we have so far identified 60 different mutations localized throughout the LDL receptor gene. In certain populations a small number of mutations predominate due to founder effects. The spectrum of LDL receptor mutations in Danish FH patients is intermediate between such specific founder populations with 5 predominant mutations (W23X, W66G, W556S, 313 + 1G-->A, 1846-1G-->A) accounting for about 40-50% of FH. These frequent mutations can easily and inexpensively be tested for by specific PCR based assays using restriction enzyme cleavage. Future analysis of LDL receptor mutations in heterozygous FH subjects, therefore, should be based on the mutational spectrum present in each relevant specific subset. Most mutations in the LDL receptor gene cause the classical heterozygous form of FH, but a small proportion seem to result in mild or moderate forms of autosomal, dominantly inherited hypercholesterolemia. Differentiation between harmless sequence variations and disease-causing mutations is not always easy without additional work. We have experienced that large re-arrangements, frame-shift and nonsense mutations obviously are pathogenic, but full pathogenicity should not be ascribed to missense mutations and small in-frame deletions, e.g. the N543H and 2393del9 mutations, unless in vitro gene expression in eukaryotic cells have been studied, or to splice-site mutations, e.g. the 1592 + 5G-->A mutation, before mRNA studies in patient cells have been performed. The cumulated LDL cholesterol exposure, mainly determined by the defect LDL receptor, plays a crucial role for the clinical manifestation of FH. The phenotypic expression of homozygous FH appears to be dominated by the consequences of the LDL receptor gene mutations. In heterozygous FH, however, the underlying mutational LDL receptor type determines only to a much lesser extent, if any, the variable phenotypic expression as seen in Danish patients. Extreme low fat dietary habits or major gene interactions may influence the lipid profile and the excess cardiovascular mortality observed in heterozygous FH, whereas minor gene determinants do not seem to play any significant role. The clinical diagnosis of heterozygous FH should be based on an elevated plasma LDL cholesterol concentration above the 95th percentiles for the general population together with either the presence of tendon xanthomas or an autosomal dominant transmission of hypercholesterolemia in the family or a child with hypercholesterolemia. Our studies illustrate clearly that molecular genetics can strengthen an equivocal clinical diagnosis and assist decision-making in diagnosis and tracing family members. If demonstration of a pathogenic mutation in the LDL receptor gene fails, other causes of autosomal dominant inherited hypercholesterolemia should be sought. Familial defective apolipoprotein B (FDB) caused by the R3500Q apolipoprotein B gene mutation may mimic FH but the clinical course, however, is often milder than that seen in patients with LDL receptor gene mutations. A newly discovered third major locus at chromosome 1 may also be of future diagnostic importance although the exact gene remains to be identified. The overall molecular genetic knowledge obtained about FH in Denmark forms the basis for the implementation and use of molecular genetic diagnostics of FH in daily clinical practice. PMID- 12553168 TI - Palliation of non-resectable carcinoma of the cardia and oesophagus by argon beam coagulation. AB - INTRODUCTION: The aim of this study was to describe the argon beam coagulation (ABC) technique in the palliative treatment of patients with nonresectable carcinoma of the cardia and oesophagus and this treatments needs, complications and tolerance. METHODOLOGY: A retrospective evaluation of 31 patients referred for palliation by ABC over a period of nearly four years (1.1.98-31.8.01). RESULTS: Twenty-two men and nine women, median age 72 years (range 49-91 years), underwent a total of 163 treatments and a median of five treatments per patient (range 1-18). Recanalisation enabling passage of the scope was achieved in 89% of treatments, and most of these patients had dysphagia grade = 2. The median range between reinterventions was 25 days (range 1-175 days). Perforation was seen in three patients and in 1.8% of treatments; procedure related mortality was 1.2%. The median hospital stay for every treatment was two days (range 1-27 days) and the median inpatient stay as a proportion of survival time was 8%. The median survival was 190 days (range 7-612 days) with the 1-year survival 19%. DISCUSSION: ABC is a well-tolerated, safe and effective treatment in patients with non-resectable cancer of the oesophagus or cardia and offers an acceptable complication rate and number of reinterventions compared with laser and stent placement. The technique is easy and cheap, and requires no further restrictions than conventional monopolar electro coagulation does. Prospective randomised trials comparing the efficacy of the different palliative treatments for patients with oesophagogastric cancer are needed. PMID- 12553169 TI - Sexual side effects induced by psychotropic drugs. AB - The majority of psychotropic drugs entail sexual side effects. The sexual side effects may reduce quality of life and may give rise to non-compliance. For example, 30-60 per cent of patients treated with antidepressants are known to develop a sexual dysfunction. However, some psychotropic drugs with no or very few sexual side effects have begun to emerge. The treatment of sexual side effects induced by psychotropic drugs may consist of: modified sexual habits, reduction in dosage, switching to another medication, possibly in combination with different psychotropic agents, other varieties of pharmacologically active substances or specific products for the treatment of sexual dysfunction such as sildenafil. Above all, it should be acknowledged that relatively few data are available in this field and in particular that there is a lack of controlled studies. PMID- 12553170 TI - Hysterectomy in Denmark. An analysis of postoperative hospitalisation, morbidity and readmission. AB - INTRODUCTION: The aim of this study was to describe the incidence of "standard" hysterectomy in Denmark, including surgical procedure, postoperative hospitalisation, morbidity, mortality and readmission rate within 30 days. MATERIAL AND METHODS: Analysis of data from the Danish National Patient Register for a two-year period (1998-2000) concerning hysterectomies for benign indications, carcinoma in situ cervicis uteri and endometrial carcinoma stage I. A stratified sample of 821 discharge resumes was reviewed for detection of complications. RESULTS: During the two years 10,171 women underwent "standard" hysterectomy followed by a median postoperative hospitalisation of 4 days. In departments performing more than 100 operations per year, the median hospital stay varied from 3 to 5.5 days. Eighty per cent were performed by laparotomy, 6% laparoscopically assisted and 14% by the vaginal route with marked regional variation in the choice of surgical approach. The number of vaginal hysterectomies varied from 0-67% in departments with a surgical activity of more than 100 per year. The mortality rate was 0.06% and 8% were readmitted within 30 days after the operation. If allowing extrapolation from the random sample to the entire population the complication rate was estimated to a minimum of 18%. CONCLUSION: "Standard" hysterectomy in Denmark is associated with considerable morbidity and marked regional variation in choice of surgical approach. The present study emphasizes the need for a national hysterectomy database with the possibility of evaluating surgical activity, reducing morbidity rates and developing homogeneous guidelines. PMID- 12553171 TI - Effects of occupational exposure to organophosphate pesticides on nerve and neuromuscular function--Peiris-John, et al. PMID- 12553172 TI - Effects of occupational exposure to organophosphate pesticides on nerve and neuromuscular function--Peiris-John, et al. PMID- 12553173 TI - Workers as research subjects: a vulnerable population--by Susan L. Rose, PhD and Charles E. Pietri, BA. PMID- 12553174 TI - The health and productivity cost burden of the "top 10" physical and mental health conditions affecting six large U.S. employers in 1999. AB - A multi-employer database that links medical, prescription drug, absence, and short term disability data at the patient level was analyzed to uncover the most costly physical and mental health conditions affecting American businesses. A unique methodology was developed involving the creation of patient episodes of care that incorporated employee productivity measures of absence and disability. Data for 374,799 employees from six large employers were analyzed. Absence and disability losses constituted 29% of the total health and productivity related expenditures for physical health conditions, and 47% for all of the mental health conditions examined. The top-10 most costly physical health conditions were: angina pectoris; essential hypertension; diabetes mellitus; mechanical low back pain; acute myocardial infarction; chronic obstructive pulmonary disease; back disorders not specified as low back; trauma to spine and spinal cord; sinusitis; and diseases of the ear, nose and throat or mastoid process. The most costly mental health disorders were: bipolar disorder, chronic maintenance; depression; depressive episode in bipolar disease; neurotic, personality and non-psychotic disorders; alcoholism;, anxiety disorders; schizophrenia, acute phase; bipolar disorders, severe mania; nonspecific neurotic, personality and non-psychotic disorders; and psychoses. Implications for employers and health plans in examining the health and productivity consequences of common health conditions are discussed. PMID- 12553175 TI - Ototoxic effects of occupational exposure to styrene and co-exposure to styrene and noise. AB - Ototoxicity of styrene and the synergistic action of styrene and noise have been shown in rats. The respective data in humans are scarce and equivocal. This study evaluated the effects of occupational exposure to styrene and combined exposures to styrene and noise on hearing. The study group, comprised of 290-yacht yard and plastic factory workers, was exposed to a mixture of organic solvents, having styrene as its main compound. The reference group, totaling 223 subjects, included (1) white-collar workers, exposed neither to solvents nor noise and (2) metal factory workers, exposed exclusively to noise. All subjects were assessed by means of a detailed questionnaire and underwent otorhinolaryngological and audiometric examinations. Multiple logistic regression analysis revealed almost a 4-fold (or 3.9; 95% CI = 2.4-6.2) increase in the odds of developing hearing loss related to styrene exposure. The factors adjusted for were: age, gender, current occupational exposure to noise, and exposure to noise in the past. In cases of the combined exposures to styrene and noise, the odds ratios were two to three times higher than the respective values for styrene-only and noise-only exposed subjects. The mean hearing thresholds--adjusted for age, gender, and exposure to noise--were significantly higher in the solvent-exposed group than in the unexposed reference group at all frequencies tested. A positive linear relationship existed between an averaged working life exposure to styrene concentration and a hearing threshold at the frequencies of 6 and 8 kHz. This study provides the epidemiological evidence that occupational exposure to styrene is related to an increased risk of hearing loss. Combined exposures to noise and styrene seem to be more ototoxic than exposure to noise alone. PMID- 12553176 TI - Influence of occupational exposure to organic solvent mixtures on contrast sensitivity in printers. AB - Contrast sensitivity measurement accurately evaluates the sensitivity of all human visual channels, even in a more detailed manner than simple acuity measures can. This is a study of visual contrast sensitivity in a series of subjects occupationally exposed to organic solvents. Contrast sensitivity was measured for 42 printers and 28 age-stratified controls using the Vistech VCTS 6500 chart. The exposed workers distinguished between following three groups of printers: "low exposed," "middle exposed," and "high exposed." Abnormal contrast sensitivity results were recorded for the right eye in 38% of the printers and 11% of the controls and for the left eye in 38% of the printers and 7% of the controls. Especially, we found a reduced contrast sensitivity in all three groups exposed to solvents for both eyes. We conclude that contrast sensitivity is abnormal in workers exposed to organic solvents. The reduced contrast sensitivity in printers seems to be an indicator of visual defects induced in response to organic solvents exposure. PMID- 12553177 TI - Higher birth weight and lower prevalence of low birth weight in children of agricultural workers than in those of workers in other occupations. AB - Agricultural occupation has been associated with different reproductive outcomes. This study examined the relation between mean birth weight (MBW) and the prevalence of low birth weight (LBW) and paternal occupation in 1995-1998. Births and late fetal deaths were analyzed (n = 1,097,817). Paternal occupation was grouped into four categories: professionals and managerial; clerical, sales and service; manual; and agricultural workers. We estimated the difference between the MBW and the prevalence ratio of LBW in each occupational group as compared to the group of professionals and managerial workers. The adjusted difference in the MBW of the children of each occupational category was -8.9 g, -17.2 g, and 16.9 g, respectively. The adjusted prevalence ratio for LBW was 1.04, 1.08, and 0.91. Our results suggest a higher birth weight and lower prevalence of LBW in children of fathers who are agricultural workers. PMID- 12553178 TI - Pesticide take-home pathway among children of agricultural workers: study design, methods, and baseline findings. AB - Farmworkers are exposed to pesticides and may take home pesticide residues to their families. In this paper, self-reported pesticide exposure and home practices to reduce the amount of pesticide residues taken home were examined among 571 farmworkers. Urine samples from a subsample of farmworkers and children and dust samples from households and vehicles also assessed pesticide exposure. Overall, 96% of respondents reported exposure to pesticides at work. Many employers did not provide resources for hand washing. Farmworkers' protective practices to keep pesticide residues out of the home were at a low level. In a subset of respondents, pesticide levels above the limit of quantitation were seen in the urine of children and adults and in house and vehicle dust. The results support the take-home pathway of pesticide exposure. Ways must be found to reduce this pesticide exposure among children of farmworkers. PMID- 12553179 TI - Effects of low-toxicity particulate matter on human nasal function. AB - Chronic nasal and paranasal sinus problems affect approximately 15% of the population in industrialized countries. Recent studies suggest that particulate matter might contribute to this condition. The effects of acute exposure to low toxicity particulate matter on human nasal airflow, mucociliary transport, and nasal discomfort should be assessed. Thirty-two healthy volunteers were exposed to 0 (control), 500, 1000, and 5000 micrograms/m3 calcium carbonate dust for 3 hours and nasal saccharin transport time (STT), rhinomanometry, and visual analog scales (VAS) on nasal discomfort were obtained. A dose dependent decrease of STT (P = 0.02) and nasal patency (P = 0.04), and increased sensation of nasal obstruction (P = 0.002) and dryness (P = 0.03) was observed. The results indicate that acute exposure to low-toxicity particulate matter in concentrations frequently encountered in western agglomeration areas may affect nasal functions and cause nasal symptoms. PMID- 12553180 TI - The changing organization of work and the safety and health of working people: a commentary. AB - Recent trends in the organization of work may affect worker health through a variety of pathways--by increasing the risk of stress-related illnesses, such as cardiovascular disease, musculoskeletal disorders, and psychological disorders, by increasing exposure to hazardous substances and violence on the job, or by affecting occupational health services and training programs. Much remains to be learned about the nature of changes in work organization, and how they affect worker health and safety. While available evidence is limited, such evidence suggests that recent trends in work organization may be increasing the risk of occupational illnesses. In a groundbreaking publication, the National Institute for Occupational Safety and Health has provided a concise summary of available knowledge and a detailed agenda for research and development. PMID- 12553181 TI - Elevated triglyceride and decreased high density lipoprotein level in carbon disulfide workers in Taiwan. AB - Carbon disulfide (CS2) is a man-made product utilized primarily in the manufacture of viscose rayon. Overexposure to CS2 has been associated with an increase in coronary heart disease. The aims of this study were to examine the dose-response relationship of CS2 exposure and elevated lipid profile tests among CS2-exposed workers in Taiwan. A total of 132 workers were recruited from two viscose rayon plants. Air sampling was performed to determine the CS2 exposure of workers. Demographic data and work history were gathered by a standard self administered questionnaire. Lipid profile tests were also performed by routine methods. The average CS2 exposure concentration was 50.6 +/- 25.6 ppm (range: 24 127 ppm) in the high-exposure group, 12.9 +/- 5 ppm (range: 5.2-22.3 ppm) in the mid-exposure group, and 3.5 +/- 1.2 ppm (range 0.97-5.2 ppm) in the low-exposure group. There were 21 out of 33 (63.7%) elevated triglyceride levels among high CS2-exposure workers, 27 out of 64 (42.2%) among the middle-CS2-exposure, and 14 out of 35 (40%) among low-CS2-exposure workers, respectively. Compared to the low CS2-exposure workers, the age- and weight-adjusted odds ratios (and 95% confidence intervals) of the prevalence of elevated triglyceride value were 1.12 (0.5, 2.7) for middle-CS2-exposure workers, and 2.81 (1.02, 7.8) for high-CS2 exposure workers. There was a significant linear trend between CS2 exposure and the prevalence of elevated triglyceride value (P = 0.046) after adjusting for other factors. There was also a lower prevalence of elevated HDL level in high CS2-exposure workers than low-CS2-exposure workers (15.2% versus 31.4%). Compared to the low-CS2-exposure workers, the age- and weight-adjusted odds ratio (and 95% confidence intervals) of elevated HDL level were 0.34 (0.1, 1.18) for high-CS2 exposure workers, which was borderline significant. In conclusion, this study suggests that elevated triglyceride level and decreased HDL level are associated with CS2 exposure. The study also suggests that exposure to CS2 (concentration > 23.2 ppm, which was above the current TLV-TWA of 10 ppm) are significantly associated with increased risk and decreased protective effect for coronary heart disease. PMID- 12553183 TI - Neurophysiological anomalies in brainstem responses of mercury-exposed children of Andean gold miners. AB - The health hazards of occupational exposure to Mercury (Hg) in adult gold miners are well known, but little attention has been given to the effects of Hg exposure in the children of gold miners. Children who assist their parents in gold mining operations or live in mining enclaves may be exposed to elemental Hg vapors or methylmercury-contaminated food, both of which may induce neurodevelopmental disabilities. Brainstem auditory-evoked responses (BAER) were measured as biomarkers of subtle mercury-induced neurological impairment in Andean children of gold miners living in the Ecuadorian gold mining settlement of Nambija, where Hg exposure is prevalent. Thirty-one children (19 boys and 12 girls, aged 4-14 years, mean age: 10 years) in the study group were found to have a mean blood mercury (HgB) level of 23.0 micrograms/L (SD: 19, range: 2.0-89.0 micrograms/L; median: 20 micrograms/L), which was significantly higher than the mean HgB level of a reference group of 21 Ecuadorian children (4.5 micrograms/L, SD: 2.3; t = 4.39, P = 0.0001), and in excess of the health-based biological limits for the U.S. (10 micrograms/L). The BAER measures indicated statistically significant differences in interpeak III-V (P = 0.03) and I-V (P = 0.008) neural conduction times for children with HgB levels above the median. BAERs at the conventional click stimulus rate of 10/second showed statistically significant positive correlations between HgB level and the absolute latency of wave V (P = 0.03), and the neural conduction times of the eighth nerve to midbrain I-V interval (P = 0.02). BAER at 50/second revealed statistically significant relationships between HgB and the latency of wave VI (P = 0.03), and the I-VI interpeak interval (P = 0.02). Brainstem neural conduction times suggested that some of the Hg intoxicated children in the study group have subtle neurophysiological anomalies that may be more manifest at higher BAER stimulus rates, and that the Hg-exposed children of gold miners are at risk for neurodevelopmental disabilities. PMID- 12553182 TI - The effects of jet fuel on immune cells of fuel system maintenance workers. AB - Jet fuel is a common occupational exposure among commercial and military maintenance workers. JP-8 jet fuel, a military formulation, has shown immunotoxic effects in mice, but little data exist for humans. The aim of this cross sectional study was to determine whether immune cell counts in the peripheral blood were altered among tank entry workers at three Air Force bases. After adjusting for covariates, fuel system maintenance personnel (n = 45) were found to have significantly higher counts of white blood cells (P = 0.01), neutrophils (P = 0.05), and monocytes (P = 0.02) when compared with a low-exposure group (n = 78), but no differences were noted in the numbers of total lymphocytes, T-cells, T-helper cells, T-suppressor cells, natural killer cells, and B-cells. Investigations are needed to evaluate the functional ability of these cells to produce lymphokines and cytokines and modulate the immune system. PMID- 12553184 TI - An exploratory investigation of suicide and occupational exposure. AB - This exploratory study evaluated the association between suicide and occupational exposure to electromagnetic fields, pesticides, and hydrocarbon solvents. The study population comprised 11,707 suicide deaths and 132,771 eligible controls identified from United States death certificate files for the years 1991 and 1992. Exposure assignment was based on job title reported on the death certificates. Exposure to electromagnetic fields and pesticides was weakly associated with suicide risk, while little evidence for an increased risk was seen for hydrocarbon solvents. The association for electromagnetic field exposure was highest for suicide between the ages 20 and 35 (odds ratio; OR = 1.5), while the highest risk of suicide for pesticide exposure was seen between the ages of 35 and 49 years (OR = 1.5). Further investigation to replicate these findings seems warranted, using higher quality occupational data. PMID- 12553185 TI - Qualifications of Medical Review Officers (MROs) in regulated and nonregulated drug testing. PMID- 12553186 TI - Pregnancy termination among South African adolescents. AB - Although African adolescents' risk of undergoing abortion and of related health complications is well-documented, little is known about the procedure's prominence in their lives and the pathways that lead to their reliance upon it. This study investigates abortion dynamics among male and female Zulu adolescents in KwaZulu/Natal, South Africa. It explores the role of abortion in young people's sexual and reproductive experience, its acceptability, the reasons and likelihood of young people's choosing abortion, and the commonly used methods of pregnancy termination. The study, a rural-urban comparison using focus-group discussions, narrative workshops, and role playing, involved surveys and in-depth interviews. Factors contributing to the commonplace nature of backstreet procedures among adolescents include: social stigma, inadequate knowledge of the legal status of abortion, and a complex group decisionmaking process. Young people invoke "relative moralities" concerning adolescent abortion, recognizing and condoning it on a context-specific basis. Age, gender, and geographic differences are examined. The methodological triangulation used offers the opportunity for alternative theoretical and methodological approaches to research on abortion-related issues. PMID- 12553187 TI - Convergence and divergence in spouses' perspectives on women's autonomy in rural India. AB - This study explores similarities and differences in the perceptions of rural Indian women and their husbands with regard to various dimensions of women's autonomy and investigates the extent to which various reproductive outcomes- contraception, unmet need, recent fertility, and spousal communication--are influenced by individual partners' views of women's autonomy. Data are drawn from a 1993-94 community-based study of women's autonomy in Uttar Pradesh and Tamil Nadu, states that are, respectively, more and less patriarchal. Matched data were obtained from 1,660 women and their husbands. Results indicate no more than a loose agreement between women and their husbands concerning the dimensions of women's autonomy within the home. Where disagreement is expressed, husbands are more likely to project a comparatively liberal picture of their wives' autonomy than do their wives, and the inference can be made that in surveys men tended to provide more "acceptable" responses than when they were questioned in greater depth. Findings also suggest that cultural context affects the influences that wives' and their husbands' perceptions of women's autonomy have on reproductive outcomes. A clear regional divide is seen, net of individual and household characteristics, in the influence of almost every aspect of women's autonomy. PMID- 12553188 TI - Son preference in a rural village in North Vietnam. AB - This article explores the continuing preference for sons in the context of low fertility in Vietnam. Although the total fertility rate for Vietnam declined from 6.0 children per woman of reproductive age in 1979 to 2.2 children in 1998, demographic evidence shows that son preference remains strong and influences contraceptive and fertility behavior. This study examines the underlying factors for son preference in a rural village in North Vietnam. The methodology includes focus-group discussions, an in-depth study of 25 families, and ethnographic observation. Results indicate that sons are highly desired for their social, symbolic, and economic value. In spite of four decades of socialist policies aimed at reducing gender-based inequalities and at weakening the patriarchal kinship system, the desire for sons continues to drive the family-building process. The article also indicates a gap between discourse and social practice with respect to roles assigned to children on the basis of their sex. PMID- 12553189 TI - Determinants of condom use among young people in urban Cameroon. AB - To reduce the prevalence of reproductive health problems among adolescents, governmental and nongovernmental organizations in Cameroon are implementing youth oriented reproductive health programs. To facilitate the design of effective programs to increase condom use, this study examines survey data on the determinants of having ever used condoms and on current condom use with regular and casual partners among unmarried young people in urban Cameroon. While most adolescents have tried condoms at least once, use remains inconsistent. Parental support, personal risk perception, and self-efficacy are found to be associated with higher levels of condom use. Youth-oriented programs seeking to increase the number of new condom users among the young should promote parental support for condom use and enhance young people's perceptions of personal risk. Programs that work to convince the young that their sexual history can put them at risk of HIV infection and that dispel the myth that HIV risk with regular partners is low may serve to increase personal risk perception. Finally, communications programs should aim to increase adolescents' self-efficacy, particularly in terms of their perceived ability to convince partners to use condoms and to use them correctly. PMID- 12553190 TI - Bangladesh 1999-2000: results from the Demographic and Health Survey. PMID- 12553191 TI - Ethiopia 2000: results from the Demographic and Health Survey. PMID- 12553192 TI - Top 10 health-care trends for 2003: rising demand, increasing revenues and innovative technologies. PMID- 12553193 TI - [Treatment of post-traumatic nail bed deformities with split-thickness nail bed grafts]. AB - INTRODUCTION: Split-thickness nail bed graft have been proposed for treatment of post-traumatic nail bed dystrophies. Only three papers reported good results in 60 to 90% of cases. We report our experience. MATERIAL AND METHODS: We reviewed 18 cases of split-thickness nail bed grafts from operative and consultation files and photographs. Evaluation was made using criteria as the nail plate shape, its adhesion on the nail bed, patient satisfaction and possible sequelae on the donor finger. RESULTANTS: Thirteen patients have been reviewed with a sufficient follow up to assess the results. In 13 cases, the graft was taken from the hallux. The nail-bed graft was used for hallux reconstruction two times, the thumb six times, index six times, middle and ring finger one each, and the little finger two times. In every patient the graft was molded using either the patient nail or the nail plate from the donor finger. Clinical results were excellent in five cases, good in three and poor in five cases. Five failures were due to wrong indications in three cases in which matrix problem has been underestimated, and infectious failures in two cases. Fifteen per cent of our patients had some sequelae on the donor finger. PMID- 12553194 TI - [Hand injuries resulting from high-pressure injection: lesions specific to industrial oil]. AB - Nineteen cases of high pressure injection injuries in the hand were treated between 1973 and 1998. Same surgical treatment plan was followed in all cases: excision of the penetration point, irrigation, debridement and synovectomy if opening of the flexor sheath was noted, and skin closure to allow early mobilisation. All cases concerned men, work injuries, and volar aspect of the hand. The elapsed time between injection and initial surgery ranged from 1 hour to 1 month with a mean of 6.5 days. Eighteen patients out of 19 were reviewed with a mean follow up of 12 years. In 11/19 cases, (58%) oil was injected. The results of oil injection cases have been analysed: the quantity of oil and the preoperative delay (if more ten hours) are associated with poor functional results or complications. Two amputations, two cases of skin necrosis at the injection point, and one case of infection are reported. One case of oleoma of the thumb is described. The specificity of injuries by industrial oil under pressure must be known: paint or white spirit are more toxic than oil which was in all cases injected in the dominant hand (no high pressure injection tool but a defect in the pipe). An important inflammatory reaction with functional sequelae is caused by foreign bodies in oil. Extraction of oil off the injured tissues is difficult because oil is not visible. A specific information is necessary for farmers and truck driver, very exposed population. PMID- 12553195 TI - Percutaneous Herbert screw fixation for fractures of the scaphoid: review of 30 cases. AB - INTRODUCTION: The purpose of this study was to evaluate the results of percutaneous fixation of undisplaced or minimally displaced fractures of the scaphoid using the first generation Herbert screw in terms of union, functional results and scaphoid mobility. METHODS: 30 of the 50 patients operated on in our department between 1995 and 2000 were available for evaluation by an independent observer. Wrist mobility, grip strength and key pinch were measured. Scaphoid mobility was evaluated by measuring radioscaphoid angles in flexed and extended positions on dynamic X-rays. RESULTS: The union rate was comparable to that achieved by non-operative management (90%). Resumption of professional activities was possible long before bony union because immobilization was short. Grip strength, wrist and scaphoid mobilities were comparable to the controlateral sides except for scaphoid flexion. Persistent symptoms were found in 30% of the patients despite union of their fracture. DISCUSSION: Our results demonstrate that percutaneous stabilization of undisplaced or minimally displaced fractures of the scaphoid preserves the mobility of the wrist and minimally alters the normal dynamics of the carpus. The duration of work inability is short. PMID- 12553196 TI - [Hand replantation in an 88-year-old man: replanted senior?]. AB - Indications of hand replantation need to take into account the patient overall condition and assess the magnitude of the injury. Although elderly people are usually considered as a limiting factor for microsurgery, indications of replantation may sometimes be extended. The author reports the case of a complete right hand amputation by a circular saw in an active right-handed 88 year man. The trans-carpal section was clean and general condition of the patient was satisfying. It was decided with the patient consent and the anaesthetist and the surgeon to perform the replantation. The procedure was carried on during 6 hours and revascularization took place 5 h 1/2 after injury. Post-operative period has been easy, but superficial nosocomial wound infection appeared which recovered with antibiotic therapy. It was not necessary to reoperate and the patient did not stay in intensive care department. After almost a 2 years follow-up, peripheral pulses were present, and the hand was warm and well coloured. Poor sensibility was found in median nerve area. Intrinsic motion was missing and extrinsic range of motion was weak. From a functional point of view, the patient was able to write, to dig his garden, and to drive his car. Analysis of this supposed hand replanted senior case report demonstrates that, when general and local conditions are safe, a hand replantation, even with a poor result, may be better than a prosthesis or an allograft, whatever the age of the patient. PMID- 12553197 TI - [Radial nerve paralysis after lateral dislocation of the elbow]. AB - Nerve injuries complicating elbow dislocation reported in literature involve ulnar and median nerve. A case of radial nerve paralysis after lateral dislocation of the elbow is reported. PMID- 12553198 TI - [Anuerysm of the ulnar artery in a mountain biker. A case report and review of the literature]. AB - A case of ulnar artery aneurysm following repeated hand injury in a mountain biker, is reported. The patient underwent surgical aneurysm resection with venous graft end-to-end anastomosis. Complete relief of symptoms is observed at 3 years follow-up. A literature review is presented. PMID- 12553199 TI - Adhesive strips: a simple and inexpensive trick for finger holding in hand surgery. AB - Many finger holding devices have been developed to retract digits and provide exposure during hand surgery. We describe a simple, and cheap trick to keep fingers out of the way using adhesive strips that has proven efficient and helpful. PMID- 12553200 TI - Evidence-based practice in action: getting started. PMID- 12553201 TI - To bridge a quality chasm: connect with the guidelines. AB - ISSUES AND PURPOSE: A wide chasm exists between the care we have and the care we could have. This article will inform nurses about the pediatric quality "chasm" by identifying practice variations from evidence-based guidelines for common childhood conditions. CONCLUSIONS: Nurses are accountable for providing state-of the-art, evidence-based care to children. The pediatric quality chasm calls for nurses to stand up and speak out for children, to use and develop tools that activate the best scientific knowledge, and to empower parents to make it happen. PRACTICE IMPLICATIONS: Advocacy and patient-centered care are nurses' means to partner with parents and collaborate with colleagues to connect with the guidelines and bridge the quality chasm. PMID- 12553202 TI - Ebb and flow when navigating adolescence: predictors of daily hassles among African-American adolescent girls. AB - ISSUES AND PURPOSE: To examine the nature of daily hassles as perceived by African-American adolescent females. DESIGN AND METHODS: As part of a larger, cross-sectional study, nonrandom network sampling technique was used to survey 178 adolescent girls between the ages of 11 and 19. RESULTS: This study found that the most common hassles were school and academic, followed by family and economic hassles, peer and social hassles, and personal safety hassles. Socioeconomic factors were strongly associated with the level of hassles reported. PRACTICE IMPLICATIONS: Assess African-American girls' perception of daily hassles, specifically school- and family-related hassles, and also examine the interrelationship between the type of hassles and health problems. PMID- 12553203 TI - Lucky girls: unintentional avoidance of adolescent pregnancy among low-income African-American females. AB - PURPOSE: To describe lucky adolescents who unintentionally avoid pregnancy. DESIGN AND METHODS: The second phase of a descriptive qualitative study in which 17 low-income African-American females ages 19 to 26 participated in open-ended interviews on how they avoided pregnancy as adolescents. RESULTS: Constant comparative analysis revealed that five of the girls avoided pregnancy because they were "lucky" that others insisted they use contraceptives. These lucky girls were unaware of sexual risks, but used contraceptives because they complied with decisions made by their parents, grandparents, and partners. PRACTICE IMPLICATIONS: Lucky girls are at risk for adolescent pregnancy because they abdicate decision making to others and are likely to be overlooked in practice because they are using contraceptives. Promoting self-protection includes assessment, knowledge, skills-building strategies, and health-promoting contracts between the nurse and adolescent. PMID- 12553204 TI - Cancer pain in children: the selection of a model to guide research. PMID- 12553205 TI - Obtaining accurate growth measurements in children. PMID- 12553206 TI - Children and the courts: advocacy roles for pediatric nurses. PMID- 12553207 TI - Leadership in IT: an unnatural but essential characteristic. PMID- 12553208 TI - New foundations for future success. PMID- 12553209 TI - The CIO as strategic thinker. PMID- 12553210 TI - The essential skills for the chief medical information officer. PMID- 12553212 TI - The Web-enabled call center: adding voice capabilities to your Web site. PMID- 12553211 TI - 2003: the year of medical paperwork simplification. PMID- 12553213 TI - What happened to the medical smart card? PMID- 12553214 TI - Key healthcare issues facing the 108th Congress. PMID- 12553215 TI - Developing a winning IT negotiations strategy. PMID- 12553216 TI - Life after go-live. Part 1: paper in the paperless practice. PMID- 12553217 TI - An interview with Stephen L. Ummel, principal, Cap Gemini Ernst & Young health consulting. Interview by Richard D. Lang. PMID- 12553218 TI - Business value of IT: the strategic planning process. AB - Too often, the vocabulary of IT justification is disconnected from the dominant business issues of the enterprise. The described framework for IT strategic planning can be used not only to align IT investments with business objectives, but also to provide clarity to enterprise business IT strategy. PMID- 12553219 TI - The digital hospital: opportunities and challenges. AB - Organizations building a new hospital have a one-time opportunity to completely rethink how care is delivered and dramatically enhance quality while concurrently reducing costs. Research clearly indicates technology is a key enabler of patient safety programs. Emerging technologies present the opportunity to automate the entire clinical continuum. PMID- 12553220 TI - Information security issues that healthcare management must understand. AB - Securing information assets is less about technology and more about business policies and procedures that managers must be involved with. Information security must be managed from a business perspective, and executive involvement is absolutely essential to ensure successful information security initiatives. PMID- 12553221 TI - Is your organization project management savvy? AB - As corporations struggle to gain better control and use of organizational resources in a rapidly changing business environment, they're finding that project management is an efficacious discipline for supporting strategic initiatives and achieving business goals. Successful project management requires an understanding of the business drivers, the organizational structure, the benefits of centralizing project management, and the factors that could impact performance. PMID- 12553222 TI - Implementing a Web-based content management system. AB - The Children's Hospital of Philadelphia transformed its web site to enhance patient satisfaction and attract new patients, as well as meet the needs of clinicians and the hospital's business plans. They accomplished this by implementing a content management system that would allow content creation and updating to be delegated to appropriate department staffers, thereby eliminating bottlenecks and unnecessary steps and ensuring that the web site receives fresh content much more quickly. PMID- 12553223 TI - Mobile physician order entry. AB - Because both computerized physician order entry (CPOE) systems and mobile technologies such as handheld devices have the potential to greatly impact the industry's future, IT vendors, hospitals, and clinicians are simply merging them into a logical convergence--"CPOE on a handheld"--with an expectation of full functionality on all platforms: computer workstations, rolling laptops, tablet PCs, and handheld devices. For these trends to succeed together, however, this expectation must be revised to establish a distinct category--mobile physician order entry (MPOE)--that is different from CPOE in form, function, and implementation. PMID- 12553224 TI - Advantages of a multi-facility information technology consortium. AB - The formation of an information technology consortium has proven to be a way for several rural hospitals in Northern Minnesota to purchase and successfully install the right technology solutions for their facilities. The concept described is very similar to that used by the Independent Grocer Alliance (IGA), which allows local independent grocers to buy products at prices comparable to these obtained by larger chain stores. PMID- 12553226 TI - Leadership--"good to great" and "built to last" by Jim Collins. PMID- 12553225 TI - IVR and administrative operations in healthcare and hospitals. AB - IVR (Interactive Voice Response) systems are effectively used in healthcare and hospitals for drug and health library information, prescription refills, history records, medical transcripts, patient information services, automated pre admissions, patient account records, appointment scheduling and reminder, lab results notification, food services, and many other applications. Among its benefits, IVR relieves administrative staff and physicians of routine customer interactions and helps healthcare organizations increase their efficiency, productivity, and profitability. PMID- 12553227 TI - OIG 2003 work plan outlines areas for compliance review. PMID- 12553228 TI - Health care: whose issue is it, anyway? PMID- 12553229 TI - Labor shortages spark innovation. An interview with Gerald F. O'Neill. AB - Labor shortages in both clinical and nonclinical areas are plaguing hospitals and health systems around the country. In Methuen, Massachusetts, Holy Family Hospital and Medical Center is pioneering new ways to fill nursing and other staff vacancies with permanent, experienced personnel who already are loyal to the hospital. Holy Family's senior vice president and CFO, Gerald F. O'Neill, discusses his organization's creative efforts to battle the healthcare labor shortage and other challenges facing healthcare organizations. PMID- 12553230 TI - Evaluating group purchasing organizations. AB - A formal evaluation process can help healthcare organizations assess the current and/or potential value of a group purchasing organization (GPO). Healthcare organizations should approach a GPO evaluation as if they were entering into a new relationship. The evaluation should include purchasing and financial services, value-added services, and corporate relations/business practices. Healthcare organizations should consider the potential economies of scale and other services offered by a GPO. Healthcare organizations should consider using acceptable substitutes for products currently used or seeking products through alternative sources if doing so achieves greater value. PMID- 12553231 TI - Exploring HIPAA revenue-cycle opportunities. PMID- 12553232 TI - Assembling the HIPAA privacy puzzle. AB - With the exception of small health plans, covered entities have less than four months to come into compliance with the final HIPAA privacy rule. Instead of obtaining written consent, providers must make a good-faith effort to obtain a patient's signed acknowledgment of receipt of the provider's notice of privacy practices. A covered entity may use patient information to market its own healthcare services without violating the privacy rule. A research institution may continue to use protected health information obtained with a valid authorization to maintain the integrity of a study even if authorization is revoked. PMID- 12553233 TI - Do you know your business associates? AB - HIPAA's business associate rules require covered entities to identify their business associates and enter into contracts with them to safeguard the privacy of individually identifiable health information. Covered entities need to determine who their business associates are and whether exceptions apply. Covered entities need to examine their business associate contracts to ensure that the contracts contain provisions required by HIPAA. Covered entities should not enter into business associate contracts unnecessarily. Existing business associate contracts are subject to a transition period for compliance with HIPAA. PMID- 12553234 TI - Case study: an Internet answer. AB - Michigan-based Munson Healthcare provided patients with access to their billing information on the Internet. Focus groups helped the provider select a system that allows patients to view their account information, check on the status of insurance or a claim, and access educational materials. Munson is receiving payment up to 26 days sooner than with paper methods, and billing inquiries from consumers have declined from 12,000 phone calls a month to 6,000. Providers should research their markets needs and weigh resource constraints when considering development of a similar program. PMID- 12553235 TI - Revenue-cycle redesign: honing the details. AB - To minimize claim denials and ensure optimum payment for delivered services, many hospitals must fundamentally change their approach to managing the revenue cycle. The revenue cycle should be conceptualized as a continuum rather than as a set of isolated events, such as submitting bills or collecting payment. Cross-functional teams composed of representatives from clinical and financial areas should be created to systematically address recurrent breakdowns detected in the revenue cycle. PFS staff should be better compensated based on their value to revenue cycle performance and receive adequate training on data collection and billing requirements to ensure the submission of clean claims. Medical-necessity screening software tools should be used at initial patient-access points to help identify scheduled services not covered by Medicare. PMID- 12553236 TI - Consultant relationships may pose liability risk. AB - Providers participating in Medicare, Medicaid, and other federal healthcare programs can be held liable for False Claims Act (FCA) violations committed by third-party consultants. No proof of intent to defraud is required to establish FCA liability. Providers can be fined triple the government's payment if they are found in violation of the FCA. Providers should examine prospective consultants' qualifications and ethical standards before entering into a contractual relationship. Providers that contract with attorneys, accountants, payment experts, and/or other third-party consultants need to monitor these relationships on an ongoing basis. PMID- 12553237 TI - HIPAA claims compliance options as easy as 1-2-3. PMID- 12553238 TI - [An in-vitro evaluation of the Sichuan Union Univ-I membrane lung and mathematical modeling calculation]. AB - A new ELF hollow fibre lung, Sichuan Union Univ-I(SUU-I), constructed in our laboratory was tested in vitro to evaluate its oxygen transfer performance before it undergoes expensive animal trial. The test rig, conditions, procedure and the modelling method are introduced in the present paper. The results demonstrate that the Sichuan Union Univ-I membrane lung possesses the gas exchange ability claimed in the AAMI standards, and approaches the levels of the widely accepted membrane lungs, MAXIMA (Medtronic, US), SARNS(3M, US) and SAFE-II (Polystan, Denmark). The in vitro technique described in this paper proved practical, economical, and simple in the design and manufacturing of a new type membrane lung, as well as in research and development of membrane lungs. PMID- 12553239 TI - [The influence of basic fibroblast growth factor on the adherence of endothelial cell to polyurethane]. AB - This study addressed the problem of how to promote the adherence of endothelial cells (EC) to polyurethane (PU). The influence of basic fibroblast growth factor (bFGF) on the adherence of EC to PU was observed by using the fluorescent method. The results showed that the attachment rates for two bFGF groups(PU precoated with fibronectin and PU precoated with collagen IV) were 71% and 68.5%, which were significantly higher than those (62.5% and 55.25%) for two control groups, respectively. These demonstrate that bFGF can promote EC attachment to PU, and suggest that bFGF can be used as a supplement factor in media for achievement of endothelialization on artificial heart and blood vessels. PMID- 12553240 TI - [In vitro testing of the intravenacaval respiratory assist device (I)--Pressure drop characteristics and oxygen transfer performance]. AB - The intravenacaval respiratory assist device(IVOX) of three different sizes developed by the present authors were evaluated experimentally in vitro. The tests using in turn two different test chamber materials(rigid and elastic) showed that a pressure drop occured. The pressure drop was higher in rigid test chamber than that in the elastic one, and the pressure drop difference between these two kinds of chamber increased with the diameter of the bundle of IVOX. Oxygen transfer rate, with water as the medium in the tests, was augmented with increasing water flow rate and gas flow rate, and with decreasing inlet oxygen concentration in water. It was concluded that, to avoid bubbles produced by IVOX, the gas flow rate should be controlled with fixed outlet water pressure, and vacancy be used in the effluent gas system. PMID- 12553241 TI - [The animal experiments and clinical trial of impeller pump used as left ventricular or biventricular assist device]. AB - The authors' impeller pump as a left ventricular or biventricular assist device was evaluated in animal experiments and clinical trial. Three calves survived over 6 weeks in left ventricular assist experiments and neither blood damage nor organ dysfunction of the animals was detected. In clinical trial, the pump supported both the greater and lesser circulation of a 6-year-old child with trilogy of Fallot and low output syndrome after open heart surgery. In 43 hours the pump worked properly. The pump weighing 240 g can perform either in pulsatile or in nonpulsatile mode. In pulsatile pumping, the impeller changes its rotating speed periodically by introducing a square wave form voltage into the motor coil. PMID- 12553242 TI - [Clinical application of hydrogel membrane of silicone rubber for preventing adhesion in orthopedics]. AB - Grafting hydrogels onto silicone rubber membranes were prepared by radiation technique for medical application. This material is characterized by high purity, hydrophilia, formation of stable hydrogel after water absorption, good biocompatibility, etc. Clinical application was initiated on the basis of animal experiments. The material was used in 47 cases of joint and tendon injuries, in 9 cases of rheumatoid arthritis, and in 4 other cases; totaling 60 cases. All patients were followed up for three and a half years after surgical operation. A general effectiveness of above 86% was noted. PMID- 12553243 TI - [The effect of activity of smooth muscles on the relationship of circumferential stress-strain of superior mesenteric veins in rabbits]. AB - The superior mesenteric veins (SMV) of 14 rabbits were divided into two groups for experiment of inflation pressure, namely the normal group and the inactivated smooth muscles group. The results showed that the relationship of circumferential stress-strain of SMV could be expressed with the exponential equation. Constant X2 is effected by the active condition of smooth muscles. The difference of the material constant X2 between the two groups was statistically significant (P < 0.05), but the difference of the material constant X1 between the two groups was not statistically significant (P > 0.05). These suggest that X2 is the parameter concerning the activity of smooth muscles. PMID- 12553244 TI - [The measurement of baroreflex sensitivity in stress-induced hypertensive rats by spectral analysis]. AB - Simultaneous spectral analysis of short-term systolic blood pressure variability (SBPV) and heart rate variability (HRV) was applied to determine the change of baroreflex sensitivity (BRS) in Sprague-Dawley (SD) rats. The modulus of the transfer function between fluctuations in systolic pressure and beat-to-beat interval would be an appropriate quantification of BRS. The animal experiment was performed on two groups of rats: normotensive rats and stress-induced hypertensive rats (SIHR). V1-receptor antagonist d(CH2)5Tye(Me) AVP of arginine vasopressin (AVP) was microinjected into intracerebroventricle. The results showed that, in base-line condition before administration, although the BRS in very low frequency band (0-0.035 Cycle/Beat, VLF), low frequency band (0.035-0.12 Cycle/Beat, LF), high frequency band (0.12-0.32 Cycle/Beat, HF) and total BRS(the sum of the three bands) were all decreased, the BRS in VLF(P < 0.05) and LF(P < 0.01) decreased significantly by statistics; and that, to the SIHR, the BRS in VLF was significantly lower after AVP-V1 administration than in base-line condition, while to normotensive rats, the BRS did not change. It indicates that the facilitating effect of AVP on the beroreflex in SIHR is mainly due to V1 receptor in central nerve system. In summary, the transfer function between SBPV and beat-to-beat interval fluctuation could be an index of BRS. This method can be developed for future clinical application. PMID- 12553245 TI - [Influence of dextran 40 and dextran 70 on the adhesion of erythrocytes to endothelial cells]. AB - Adopting the flow chamber realtime microrecordation system, we studied the dynamic effects of dextran 40(DX40) and dextran 70(DX70) on the adhesion of erythrocytes to monolayer of cultured human umbilical vein endothelial cells (HUVEC). The results demonstrated that erytherocytes pretreated with DX40 adhered to HUVEC less than the erythrocytes of the control group did, but DX70 increased the number of erythrocytes adhering to HUVEC. The higher the concentration of DX70 was, the more the erythrocytes pretreated with DX70 adhered to HUVEC. When shear stress increased, the decrease rate of the number of erythrocytes (pretreated with DX70) adhering to HUVEC declined with the increase of DX70 concentration. These indicate that DX40 as an expander of blood plasma is better than DX70. PMID- 12553246 TI - [A study of non-steady flow properties in flow chamber experiment]. AB - A time-dependent velocity equation has been achieved by means of eigenfunction method in this paper, which can satisfy any impetus and describe the changing process from the initial time to any time. The theoretical basis is provided to study the nature of blood cells in non-steady flow. PMID- 12553247 TI - [Correlation noise detection in encephaloblood-fluidity signal]. AB - The aim of this study was to remove random white noise which mixes with encephaloblood fluidity signal. Based on the principles of correlation detection and a priori knowledge of encephaloblood fluidity, the authors adopted the coherent averaging method and obtained pure encephaloblood fluidity signal. The result of practical use has demonstrated that the more times the averaging progresses, the more distinctly the wave shows up. PMID- 12553248 TI - [Application of constrained simulated annealing to 3D stereotactic radiotherapy treatment planning]. AB - In this paper, linear programming and constrained simulated annealing are combined to optimize X-knife stereotactic radiotherapy treatment planning. In the first phase, linear programming reduces the resolution space, which is favourable to the adoption of little granularity in the second phase. Our experiment demonstrates that after the two-phase optimization, we attain the expected result. PMID- 12553250 TI - [The use of isoparametric element of arbitrary hexahedron for solving 3-D thoracic potential distribution]. AB - Isoparametric element of arbitrary hexahedron plays an important role in finite element analysis. This article devotes itself to establishment of mathematical model and combination of derivative of shape function of such element type. In addition, the article also discusses the usage of isoparametric element for solving 3-D thoracic potential distribution in electrical field. PMID- 12553249 TI - [A methodological study of frequency domain analysis on heart rate variability and RT interval variability]. AB - Recently, heart rate variability (HRV) as a non-invasive measure for assessing the balance of autonomic nervous system has drawn more attentions and HRV analysis has become a focus of the ECG signal processing. QT interval represents the ventricular repolarization duration. QTV might reflect the balance of autonomic nervous system in other way. QTV analysis might complement HRV analysis. Because of the difficulty in detecting QT interval, RT interval is often used instead of QT interval. HRV and QTV detecting methods have been presented in this paper. After recording 24 h ECG signals, R fiducial marks are detected correctly and ectopic beats are removed. T peaks are detected on the basis of templet matching and correlation. The analysis in frequency domain consists of 24 h 3-dimensional frequency chart, as well as trends of HF/LF and HF, with fast Fourier transform and autoregressive techniques. We acquired ECG signals from subjects under controlled respiration in different frequencies. The results of HRV and RTV spectral analysis have shown that HF peak is modulated at the frequency of controlled respiration, which proves that HF peak is dependent on the vagal activity and our method is correct. The establishment of these methods has provided a basis for further research, and the development of HRV and QTV analysis techniques will be more helpful for early diagnosis, monitoring and prognostic assessment of some diseases. PMID- 12553251 TI - [Clustering analysis of phonocardiagrams]. AB - Through the development of the ART-2 neural network, we had constructed the 2 dimensional ART neural network appropriate for clustering the time-varying spectrums. After that, according to the character of phonocardiagram(PCG), we chose the scalogram of PCG to be the input feature of the network. By adjusting parameters the network was enabled to cluster the input PCGs to two clusters. The result demonstrates that the system we designed can pick out, from among 32 continuous PCGs, two PCGs which have the tendency toward abnormality. This result may be applicable to basic physiological studies and clinical diagnosis. PMID- 12553252 TI - [The spectral broadening correction in peak blood flow velocity estimation]. AB - Numerous effects contribute to the ultimate shape of output power spectrum in Doppler ultrasound blood flow measurement. The output Doppler spectrum is broadened by physiological sources related to blood flow velocity as well as some unexpected broaden components due to the nonideal acoustical factors of the ultrasound unit. Therefore, by using the classical Doppler equation, it will lead to overestimation of the maximum velocity in conventional Doppler blood flow measurement. In this paper, we present a modified Doppler equation which takes the intrinsic spectral broadening into account. The foundation of this method is introduced and the results of the in vivo experiment using this corrected equation are reported. PMID- 12553253 TI - [Studies on target localization of Parkinson's disease treated with gamma knife]. AB - Ninety cases of Parkinson's disease were treated in our center from Dec. 1993 to Aug. 1997. Based on our work, we have developed a method for the MR localization in the treatment of Parkinson's disease with Gamma knife, including the fixing of stereotactic frame, the selection of target, and the correction of the target's coordinates. In this paper, the effects of fixing of coordinate frame on the localization of target have been analyzed. Also presented are the errors of the coordinates of the selected targets, compared with those corrected. The method reflects a new idea for target localization. PMID- 12553254 TI - [Assessment of diagnostic result for medical image by receiver operating characteristic and forced choice]. AB - Two methods for diagnostic image evaluation, the receiver operating characteristic (ROC) and the forced choice, are introduced in this paper. The concepts of ROC and forced choice are described firstly, then the experiment design consideration and data analysis are discussed. PMID- 12553255 TI - [A robust method for automated retinal vascular images mosaic based on phase correlation method and mathematical morphology]. AB - Precise mosaic techniques are essential for quantitative evaluation of retinal images to make early detection of fundus anomalies feasible. Opening of a gray scale image by a gray-scale structuring element(SE) can generate a background image. Image mosaic was achieved by subtraction this background image from the original image and then applying a phase correlation method to find translation difference. Because of the accuracy characteristics of the phase correlation method and the speed of the FFT hardware, this new algorithm can work very fast and accurately, compared to conventional techniques. The method was also characterized by an outstanding robustness against correlated noise and disturbances, such as those encountered with nonuniform illumination. The results confirm the robustness of the chosen approach. PMID- 12553256 TI - [Hyaluronic acid and laminin in patients with ischemic necrosis of femoral head]. AB - To study the course and clinical significance of hyaluronic acid (HA) and laminin(LN), the main components of extracellular matrix in ischemic necrosis of femoral head(INFH), the quantities of HA and LN were measured by radioimmunoassay in 45 patients suffering from INFH with different pathogenesis, and in 30 normal subjects(NS). The results showed that the level of HA in patients with INFH was markedly higher than that of NS group(t = 3.29, P < 0.01). The patients with hormone type INFH had the highest level of HA(t = 3.62, P < 0.01). The level of LN in patient with INFH was significantly higher than that of NS group(t = 2.84, P < 0.01). The increased levels of HA and LN were positively correlated with the development of the disease. These suggest that quantity analysis of HA and LN may be used as a good indicator in early diagnosis and prognosis of INFH. PMID- 12553257 TI - [Mechanism of specific effects of transient electromagnetic pulses on cell model]. AB - In this paper, a facilitated cell model has been built to analyze the electromagnetic pulses (EMP) induced transmembrane potentials. The analysis shows that the athermal biological effects are caused by micropores on the membrance, not directly by transmembrane potentials. These micropores are enlarged by weak transient EMP and finally lead to membrane openings to make ions cross. The electroporation effects are used to explain the specific response of biological cells to transient EMP. PMID- 12553258 TI - [Pulsatile microdialysis method with glucose sensor system]. AB - To develop a miniature subcutaneous glucose sensor system, to simplify calibration procedure before or after glucose monitor, and to improve its temperature characteristics, a pulsatile micropump and the microdialysis technique were used. The experiments of glucose recovery in 5.5 mM and 10 mM G.S. and the temperature experiment from 20 degrees C to 40 degrees C were conducted. The results showed that glucose recovery was (97 +/- 2)%, the current values of glucose sensor varied with temperature, but the temperature parameter was only 3.4%. These indicate that the glucose sensor system is miniature and the calibration procedure is simple. Because of its modified temperature characterisic, it is useful to monitor the blood glucose of feverish diabetics. PMID- 12553259 TI - [Dental welding titanium and its clinical usage]. AB - Due to its excellent biocompatibility, desirable chemical and mechanical properties, Titanium has been used for implant denture, RPD and FPD, where welding techniques were indispensable. This paper introduces 5 useful modern ways to weld Titanium and their clinical usage. They are: laser, plasma welding, TIG, infraned brazing and Hruska electrowelding. PMID- 12553260 TI - [A novel high quality specific antithrombotic drug--hirudin]. AB - Hirudin is the most potent specific inhibitor of thrombin, which is currently undergoing clinical trials as a novel antithrombotic drug. In this paper, the composition and structure, the pharmacological and pharmacokinetic characteristics, the anticoagulant and antithrombotic activity and function, the methods of isolation, recombination and syntheses were reviewed. PMID- 12553261 TI - [Development condition of three dimensional ultrasonic imaging and analysis of some key technologies]. AB - This paper introduces the value of three-dimensional ultrasonic imaging and it's different realizing approaches. It deliberately analyzes the key techniques used in three-dimensional ultrasonic imaging. These techniques include registration of two-dimensional images, accurate reconstruction of three-dimensional volume, projection of irregularly sampled plane and segmentation of three-dimensional image. The development status and future trend are also given in this paper. PMID- 12553262 TI - [The real-time classification of motor unit potentials]. AB - Motor unit potentials(MUP) are real-time classified with the method of correlation in this paper. The correlation coefficients are sorted to reserve the better waves of MUP. Not only the store waves and analytical time of MUP can be reduced, but also the measurement correction and precision of MUP parameters can be improved. PMID- 12553263 TI - [A study of cytomorphology by means of computer-aided quantitative analysis]. PMID- 12553264 TI - [Preparation of latices with functional group containing N and their application in rapid diagnosis of some diseases]. AB - This article reports the preparation and diagnostic application of a variety of latices. These latices, including temary copolymer diazo latex, azidocarbonyl latex and cell labeling latex are stable, monodisperse, soap-free, and covalently combined with protein in the absence of condensing agent. We have improved diazo polystyrene latex and made ternary copolymer diazo latex for early and rapid diagnosis of some diseases. Immunoassay reagent for the dectection of dengue fever has been prepared by our lab using ternary copolymeric diazo latex and azidocarbonyl latex. The experiment showed virus antigen could be detected in 5 mins by using diazo diagnostic agent. The sensitivity of detecting patient serum sample was 82.25%; no-across reaction was observed in other inserted spread virus antigens and in serum of other feverish patient and normal control. 40 serum samples were dectected by latex agglutination assay using ternary copolymeric diazo latex immunoassay reagent. The results were compared with those of PCR and virus separation methods. The sensitivities of the three methods were 82.25%, 75% and 55% respectively; the combined sensitivity was 92.5%; higher sensitivities could be obtained by the former two methods within 1-13 days after the onset of disease. PMID- 12553266 TI - [Analyses on structural and physicochemical properties of collagen hemostatic sponge]. AB - The ultraviolet spectrum and aminoacid content of collagen hemostatic sponge prepared from bovine tendon were analyzed, and the struture and composition of collagen spoge were confirmed. Physicochemical properties measurements showed that collagen hemostatic sponge had excellently stable quality. It was concluded that the technology for preparing collagen hemostatic sponge was reasonable and reliable. PMID- 12553265 TI - [A comparative study on cytocompatibility of medical PVA and intelligent PVA-g NIPAAm hydrogels]. AB - "Intelligent" Polymers which resonse with large property changes to small physical or chemical stimuli are attracting more and more attention in the field of medical science. One trend in the area is to modify some conventional medical polymers to form intelligent biomaterials. This investigation was designed to evaluate and compare in vitro cytocompatibility of PVA hydrogel, which could be used as a kind of solid support with good biomcompatibility, and PVA-g-NIPAAm, which was produced by irradiation grafting of NIPAAm onto the said PVA hydrogel support. Cytotoxicity test was carried out with a modified Cell Relative Growth Rate test (RGR test). On the day 2, day 4 and day 7, the RGRs of the PVA hydrogel were 82%, 90% and 102% respectively while the RGRs of the PVA-g-NIPAAm were 77%, 86% and 100%. The grades of the cytotoxicity of the PVA and PVA-g-NIPAAm hydrogels were 0, so they had no cytotoxicity as judged by our tests. After 24 hours, cells in direct contact with the two kinds of samples were observed morphologically through the Olympus inverted microscope and had a normal development. The conclusion suggests that irradiation grafting is a good way to modify conventional medical polymers to produce intelligent biomaterials. PMID- 12553267 TI - [The development of a kind of artificial articular cartilage-PVA-hydrogel]. AB - In this study, the process included aqueous Poly (vinyl alcohol) solutions frozen at -20 degrees C for 6-12 h, and then thawed for 1-2 h. The same process was repeated 1-3 times. After the specimen was dehydrated in vacuum, a kind of artificial articular cartilage--PVA-hydrogel was made up. Subsequently, the micromorphology of PVA-hydrogel was observed by means of optical microscopy and SEM. DSC and mechanical tests were employed in ivestigating the influence of freesing, thawing, dehydrating and irradiating upon the crystallity and the mechanical properties of PVA-hydrogel. PMID- 12553268 TI - [Evaluation on cytotoxicity of gallium alloy]. AB - Cytotoxicity is an important indicator to the biocompatibility of biomaterials. In this study, the cytotoxicity of Gallium alloy was evaluated by the method of cell relative growth rate in vitrousing L-929 mouse fibroblast. The cell relative growth rate of Gallium alloy was the absorbency ratio between the solution of Gallium alloy and negative control. The relative growth rates of Gallium aloy were converted into the scores of cytotoxicity. It was found from the results that Gallium alloy had no cytotoxicity. PMID- 12553269 TI - [Three-dimensional finite element study on periodontal membrane of central maxillary incisor under physiological load]. AB - This paper presents the development of a three-dimensional finite element model of central maxillary incisor. Stress distribution of the periodontal membrane was obtained by calculation under physiological load exerted to the natural tooth and tooth restored with post respectively. And it was compared with the stress distribution of the implate (contact with bone and without periodontal membrane). The result indicated that, under physiological load, when the natural tooth was compared with tooth restored with post, there were no apparent diferences in stress distribution of periodontal membrane and in stress values on inner surface and outer surface of periodontal membrane. It suggests that periodontal membrane has apparent cushioning action against stress. PMID- 12553270 TI - [A biomechanical study on human lataral capsular-ligament complex of temporomandibular joint]. AB - Tensile mechanical experiments were made on 31 lateral capsular-ligament complex spencimens from 8 fresh temporomandibular joints of human cadavers aged 8-15 years. The tests included stress relaxation at different strain rates and tension test at constant strain rate. Some biomechanical material properties of the tissue were determined. The result showed that the complex had lower tensile strength and tensile stiffness, and the stress relaxatioin was the most important mechanism of spreading stress and absorbing shock. The stress relaxation function and the constitutive equation of stress and strain were obtained with Fung's Quasi-Linear Viscoelastic Theory fitting the experimental results. PMID- 12553271 TI - [Mechanism study of muscle injury from high tension (I)--The biomechanical effect of training on muscle injury from downhill running in rats]. AB - In this experiment, we tested the biomechanical characteristics of rat's fast and slow skeletal injury from downhill running at different times. The result indicates that the mechanical properties of fast and slow skeletal muscle, including maximum tensile strength, maximum strain, elasticity and energy absorption, gradually stregthen with extended time (not more than 48 hours) after eccentric exercise-induced injury. On the other hand, the maximum strain of slow muscle is higher than that of fast muscle. PMID- 12553272 TI - [Primary study on HRV analysis of nonlinear dynamics in animal experiment]. AB - The purpose of this study was to address the relationship between the parameters of nonlinear dynamics and functions of heart. We adopted the methods of nonlinear dynamics in observing the changes of the parameters of nonlinear dynamics under the conditions that sympathetic or parasympathetic nerve of heart exists singly as well as the coronary artery is narrowed. The results showed that in case the sympathetic or parasympathetic nerve was cut off the correlation dimension and entropy were lower than the normals, and in case the coronary artery was seriously narrowed the correlation dimension, entropy and laypunav exponent were also lower than the normals. The conclusion is that the parameters of nolinear dynamics can indicate the changes of the functions of autonomic nerve under the above conditions. PMID- 12553273 TI - [Using complexity measurement to study the EEG signal of focal ischemic cerebral injury]. AB - In this paper, a model for SD rat focal ischemic cerebral injury is presented. Based on this experiment model, we recorded the EEG of ischemic region and normal region from before ischemia to after ischemia 30 minutes. We analyzed the complexity of these EEG signals, and found that the EEG complexity was very sensitive to focal ischemic cerebral injury. The EEG complexity on ischemic region was less than that on normal region. The difference in EEG complexity between two regions increased with the decreasing velocity. When insult took place, the EEG complexity was less than that before ischemai. These resluts agree to those we obtained by Discrete Gabor Spectrogram analysis and Heat Shock Protein (HSP) expression. The ischemic and normal regions can be clearly distinguished by analyzing their EEG complexity, and the extent can also be detected in this way. PMID- 12553274 TI - [Blood pressure examination using oscillometric method]. AB - The effect of pressure sensor, filter and blood pressure algorithm upon oscillometric method is discussed in this paper. The design principle and parameters configuration used in portable blood pressure HOLTER are presented. PMID- 12553275 TI - [The measurement of QT dispersion using wavelet transformation]. AB - This paper addressed the application of wavelet to the measurement of QT dispersion and presented a new method using multi-thresholds to determinate the end point of T-wave. The results geined by computer and human were compared. We also suggested that QT dispersion be defined as the SD of 12 QT intervals. PMID- 12553276 TI - [The characteristic wave intervals of exercise ECG influenced by changing of heart rate]. AB - This paper analyzes the characteristic wave intervals of exercise ECG influenced by heart rate changing under exercise test. The most influenced parameter is TP interval, the next is QT interval, and the width of P wave and QRS waves are nearly unchanging. According to the changing characteristic of these waves width mentioned above, we suggest an adaptive adjusting method for ST segment measuring position after the J point, which is of significance to recognizing the characteristic waves and determining the result of exercise test for myocardial ischemia. PMID- 12553277 TI - [Analysis of Lyapunov exponents of heart rate variability in diabetics]. AB - This paper presents an analysis of Lyapunov exponents of ten normal persons and ten patients with diabetes mellitus by the definition of the Lyapunov exponents, the length evolution and the area evolution. The results show that the Lyapunov exponent of diabetic is clearly lower than that of normal. It can indicate the diabetic with less number of electrocardiographic data. PMID- 12553278 TI - [Determination of AR parameters for multi-channel upper limb EMG]. AB - In this study, a segment LS method was used to determine the AR parameters for multi-channel upper limb EMG. Different sampling frequencies, time-windows and attenuation factors were tested and an optimum selection was obtained. According to the experiment result, the segment LS algorithm should be the best method compared with others. PMID- 12553279 TI - [Electrically evoked otoacoustic emissions and their distortion products in guinea pigs]. AB - The objective of this study was to investigate micromechanical properties and electromotility of cochlear outer hair cells in vivo. Extracochlear electrically stimulation with sinusoid alternating current was delivered to cochlea of guinea pigs. Sound pressure level was recorded from ear canal by microphone and the electrically evoked otoacoustic emissions (EEOE) and their distortion products (DPEEOE) were analyzed with FFT spectrum analyzer. The EEOE in 3 kHz to 33 kHz were recorded from 15 guinea-pigs in 18 guinea-pigs, the transfer function was more smooth in 8 kHz to 31 kHz. The DP EEOE were very clear when F1 = 6 kHz, F2 = 7.2 kHz, and the current intensity higher than 100 microA. While the intensity increased to 300 microA, two distortion products, F2-F1 and 2(2F1-F2)-F1, could be seen in addition to 2F1-F2. The input-output function showed that EEOE and DP EEOE I/O function were linear when lower electric intensity stimulation was given, but they displayed compress non-linear features when higher intensity current was delivered. The authors conclude that EEOE and DP EEOE are expression of electrophonic hearing characteriged by wide frequency bandpass appearance, wide dynamic range and non-linear features, so they are good tools for studying the mechanical properties of outer hair cells and the integrate function of Corti's organ. PMID- 12553280 TI - [The effect of age on the regional blood flow of neck of femur in normal Chinese]. AB - This study aimed to assess the effect of age on the regional blood flow of neck of femur in normal Chinese. 133Xe washout method was adopted. The result showed that the femur regional blood flow of normal Chinese decreased slowly with the increase of their age. The relation between age and femur blood flow might be expressed as F = -0.042Y + 9.01(F refers to blood flow, Y refers to age). The average femur regional blood flow of normal Chinese was 7.37 +/- 0.62 ml.(100 g.min)-1 for 21-40 years old. The authors suggest this result can be considered to stand for regional femur blood flow of young healthy adults. PMID- 12553281 TI - [A new viscoelastic model for granulocytes]. AB - In this paper, a new viscoelastic model for granulocytes was introduced. The stress was placed on the theory and its experimental results of three-parameter viscous model with micropipette aspirating technique. The small deformation of single rat granulocyte into the micropipette was analyzed by the model. Within the small deformation, the fit of the experiment data using the three-parameter viscous model was more satisfactory than that using the standard solid model, and the former suggested that cytoplasm of granulocyte was treated as a liquid rather than a solid. PMID- 12553282 TI - [A robust method for automated detection and grading of retinal venous beading in diabetes mellitus]. AB - Diabetes mellitus is a high prevalence disease in the world and retinal vessel changes such as venous beading are common findings in patients suffering from long-standing diabetes. Three methods (1. Combined use of the SED, ROSE, Modified LoG algorithm; 2. Gaussian fitting-based method; 3. Gradient-based method) for detection venous caliber were used in the first processing stage, then the magnitude of fast Fourier transform of variation function of venous caliber was used as a detective index of venous beading. The reliability of the results, justified by their low variability, makes feasible the standardization of quantitative parameters to be used both in the diagnosis and in the prognosis of diabetes, and also allows prospective studies based upon them. PMID- 12553283 TI - [Analysis of the relationship between the facial skin temperature and blood flow]. AB - The relationship between the blood flow of temperal or facial artery and facial skin temperature was studied in 30 patients with facial nerve paralysis. The facial skin temperature was recorded by infrared thermography, and the blood flow was measured by ultrasonic Doppler flowetry. The results showed, in the same facial region, the greater the volume of blood flow was, the higher the skin temperature stood. After acupuncture on the facial points, the facial blood flow increased and the facial skin temperature rose. The ratios between the increase in facial skin temperature and the in increase in blood flow were 0.9538 1.2835(degrees:ml/second) in four facial regions. These indicate the presence of certain quantitative relationship between the facial blood flow and the skin temperature. PMID- 12553284 TI - [The research of evaluation the compatibility of biotic material in cell culturing method]. AB - It is quick convienent good-repeating and cheap that examining the biotic material's compatibility through cell-culturing method, and it is more and more important in evaluating the compatibility of biotic material. The new material appearing continously complicating of the part and aim material be planted in the intensity of material's toxic effect the reaction's complication of material and biotic body, all of these decide the variety of experiment method and cells in cell toxicity experiment. It is very important that choices the right experiment method and cells according to the material's character the part and aim the material be planted in. The evaluation of biotic material's compatibility stressed on the changing of cell's form and quantity before. In recent years, more and more reports appear about material influences the growth. Adhesion proliferation and metabolizing of cell, and presents the point that the evaluation standard of biotic material's compatibility should be set according to the active cell's quantity and their growth. Combining many subject's technological development, such as immunology, chemistry, radiation and shadowgraphy, thoroughly inquires the changing relation of cell's structure and function, furtherly clarifes the material's effect on cell. It is the developing direction in the future that evaluates the biotic material's compatibility in cell-culturing method. PMID- 12553285 TI - [The advances of research for problems of developing flow in blood vessel]. AB - The development and the tendency of the problems of developing flow in the vessel are reviewed in this paper. It is pointed out that the problems of developing flow of viscoelastic blood vessel are front subjects, and that consideration shouldbe be give to three aspects, namely the blood flow, the blood vessel's wall motion and the coupling between the blood and the wall. PMID- 12553286 TI - [Quantitative backscatter imaging]. AB - Some methods for producing quantitative backscatter images are presented. As a basis, the data reduction method calculates absolute coefficient. The reference phantom method is easier to implement for clinical application. An application method is also presented which separates the contributions of scatter size and scattering strength to backscatter signal and displays Size image and Strength image separately. This method is generally accepted by the medical community. At last, the recent development of ultrasonic backscatter imaging is reviewed. PMID- 12553287 TI - [The present research situation of cause of formation of impedance cardiograph waveform]. AB - This paper summarized the present research situation of cause of formation of impedance cardiograph waveform in two aspects of early and current research on it by the means of consulting a great deal of domestic and foreign data. The paper pointed out that a practicable way for correcting Kubicek theory and its physical model, making SV formula more accurate is to apply finite element technology to the research field under the guide of impedance cardiograph electric field theory. At the same time, the paper made preliminary introduction on the author's research work in the area. PMID- 12553288 TI - [Research highlight of image reconstruction today]. AB - In today's image processing researches, image reconstruction is a highlight. It includes the following several aspects: (1) reducing noise before reconstruction or after reconstruction under time-frequency constraints; (2) researches for reconstruction of motion organ; (3) image reconstruction from incomplete projection data; (4) direct volume reconstruction and 3-D image rendering; (5) researches for non-X-ray CT; (6) researches for parallel processing system of CT. In this paper, an introduction to these aspects is given. PMID- 12553289 TI - [Visible human data set and it's application]. AB - The Visible Human Project is an outgrowth of the NLM's 1986 Long-Range Pian. It is creating a complete, anatomically detailed, three-dimensional representations of the male and female human body. This paper makes a brief description about VHP and it's significance, how to get VH data set and how to use it. PMID- 12553290 TI - [Limited diffraction beams and its application]. AB - Limited diffraction beams have a large depth of field and thus have many applications such as medical imaging, tissue characterization, material flaw detection and blood flow detection by Doppler method. Nondiffraction beams colud be produced only by transducer with an infinity large aperture in theory. The experimental results show that limited diffraction beams produced by a limited aperture have desirable large depth of field also. The theory of this kind of beams and their applications in field of medical imaging and blood flow estimation are introduced. PMID- 12553291 TI - [Preliminary study of a new concept for left ventricular assistance dynamic aortic valve]. AB - Dynamic aortic valve, a novel design for left ventricular assistance is proposed. An impeller similar to the airscrew is placed in the position of aortic valve. The impeller is held by a pair of bearings in the center of a cylindrical supporting cage which is located in the lumen of the aorta ascendens. The supporting cage is attached to the valve annulus by a sewing ring of textile cuff around the outside of the cage. A rotor is fitted into the cage and connected to the impeller with a common shaft. The cage supports the aortic wall, thus prevents the rotor from founching the wall and allows the rotor to rotate freely. A stator, functioning as that of the electric motor, is located outside the aortic wall. When actuated, the magnetic field generated by the stator would drive the rotor to turning, and the axial flow of blood from ventricular to the aorta would be produced, resulting in the reduction of the ventricular load. We studied this concept in a simulating model. The result suggests that this design be reasonable. Further exploration of this concept is justified. PMID- 12553292 TI - [The design of an ultrasonics Doppler spectrum analysis system for fetal umbilical blood flow monitoring]. AB - This paper explains the design of an Ultrasonics Doppler Spectrum Analysis System for fetal umbilical blood flow monitoring. The gist of discussion includes the sampling method of the Doppler signals and the algorithm of the blood flow parameters. Because the DSP chip is not used in the AD cardboard, the hardware design is simplified and the system works steadily. In the process of computing the blood flow paramenters, an autochange method of the amplitude threshold and a timeout control technology are used, so the algorithm has good adaptability and works very stable. PMID- 12553293 TI - [The establishment of prescription dose system for radiotherapy on accelerator's X-ray]. AB - This study aimed at how to calculate the prescription dose quickly and accurately. According to the conditions of radiotherapy and the dose of target point. RT-100 Radiotherapy Dosemetre, which was calibrated at the Industrial Health Institute of National Ministry of Health, was used to detect the dose distribution of special points in the phantom. Functions of dose distribution were fitted on the basis of the detected points. The dose ratio was introduced, and the calculation protocol indexed with Chinese for prescription dose was programmed. The doses of assumed points at the phantom which imitated various conditions of radiotherapy were detected. The detected doses were very close to the calculated doses, with the relative error less than 1.2%. It is concluded that dose of target point can be transformed to the prescription dose quickly and accurately with the method of dose ratio. PMID- 12553294 TI - Hip revisions with impacted morselized allograft bone and cement. Patient outcome, prosthetic fixation and risks. PMID- 12553295 TI - Role of the oocyte nucleus in determination of the dorsoventral polarity of Drosophila as revealed by molecular analysis of the K10 gene. AB - In Drosophila, the establishment of dorsoventral polarity of the developing embryo depends on the expression of at least 11 maternally acting genes. Mutant females that lack any of these gene activities produce normally shaped eggs that develop into dorsalized embryos. The female sterile K10 mutation differs from these mutants, because in addition to the dorsalized development of the embryo, it causes a dorsalization of the egg shape. During oogenesis, the K10 gene is specifically expressed in the oocyte. Antibodies raised against a beta galactosidase-K10 fusion protein were used to visualize the K10 product in ovaries by indirect immunofluorescence. The protein, which contains a putative DNA recognition helix, accumulates in the nucleus of the oocyte, where it is assumed to have a regulatory function. Our results thus indicate that the controlled expression of some of the genes of the oocyte nucleus is essential for the determination of the dorsoventral polarity of the oocyte and possibility of the developing embryo. PMID- 12553296 TI - Medicaid managed care and working-age beneficiaries with disabilities and chronic illnesses. AB - Even after a decade of study, there are still many questions about how working age adults with disabilities or chronic illnesses are faring in Medicaid managed care. This article draws from recent research to examine two critical issues: the need for better outcome measures specific to this population and the importance of evaluations that accurately estimate program effects, despite continued policy change, considerable differences among States, and extreme population diversity. The authors outline a research agenda that will help policymakers develop a more precise sense of how managed care practices affect different subgroups of working age adults with disabilities and chronic illnesses and how practices could be altered to improve Medicaid's ability to meet their needs. PMID- 12553297 TI - What does voluntary disenrollment from Medicare+Choice plans mean to beneficiaries? AB - The Balanced Budget Act (BBA) of 1997 required CMS to report publicly Medicare managed care (MMC) plan voluntary disenrollment rates. To ensure disenrollment rates would be meaningful to beneficiaries in health plan choice, CMS funded the development of surveys and reporting formats to identify and present the reasons that beneficiaries voluntarily leave plans. Public reporting of reasons on the Medicare Web site began in 2002. We discuss results from extensive audience testing of disenrollment rates and reasons materials. Medicare beneficiaries do not easily understand disenrollment. We also discuss challenges in presenting useful disenrollment information and policy implications for public reporting. PMID- 12553298 TI - Whole-body cooling after perinatal asphyxia: a pilot study in term neonates. AB - In order to test the practicability and safety of whole-body cooling in term neonates with moderate-to-severe hypoxic-ischaemic encephalopathy (HIE) and to report outcomes, a prospective pilot study was carried out in 25 term infants (median postmenstrual age 38 weeks, range 36 to 41 weeks; 20 males, five females). Whole-body cooling, to a target core temperature of 33 to 34 degrees C, started within 6 hours of birth and was maintained for 72 hours. Of the 25 newborn infants (19 Sarnat II and six Sarnat III, 18 outborn), 18 survived, including 13 (72%) with normal cerebral signal by MRI. Temperature instability occurred during cooling in 15 infants, but neither severe haemodynamic instability nor renal failure was seen. Thrombocytopenia developed in 12 infants, including seven with biological disseminated intravascular coagulation. One patient had hypoxaemia with right-to-left shunting through the ductus arteriosus, and seven had limited meningeal or subdural bleeding. Whole-body cooling is feasible in term neonates, with no life-threatening adverse events. Improvements are needed to obtain stable hypothermia for 72 hours. PMID- 12553299 TI - EuSEM core curriculum for emergency medicine. PMID- 12553300 TI - Influenza-like symptoms and thrombocytopenia in a teenager. PMID- 12553301 TI - Congenital cytomegalovirus infection in San Luis Potosi, Mexico. AB - The incidence of congenital cytomegalovirus infection in Mexico is unknown. We evaluated the presence of cytomegalovirus infection in 560 newborn infants at a public general hospital. There were five (0.89%) infected newborns. Infants with congenital infection were more likely to be born to primigravid mothers (P = 0.01) and were more often from rural areas (P = 0.058) than were noninfected newborns. PMID- 12553302 TI - Simultaneous infection with Borrelia burgdorferi and human granulocytic ehrlichiosis. AB - A 5-year-old child with simultaneous early Lyme disease and human granulocytic ehrlichiosis (HGE) is described. Because of the shared vector, HGE and Lyme disease are increasingly identified as coinfections in tick-exposed patients. Early recognition of concurrent Lyme disease and HGE is important because amoxicillin, an antibiotic of choice for young children with early Lyme disease, is ineffective for HGE. PMID- 12553303 TI - Presence of the new human metapneumovirus in French children with bronchiolitis. AB - We developed a polymerase chain reaction (PCR) to detect human metapneumovirus (hMPV) in French children hospitalized for acute respiratory tract disease during the winter of 2001 to 2002. This assay showed that 19 (6.6%) of 337 respiratory specimens that were negative for other respiratory viruses were positive for hMPV. Seven hMPV infections were also detected in the previous winter. The patients had the following clinical syndromes: bronchiolitis, 16; acute exacerbation of asthma, 4; bronchitis, 1; laryngitis, 1; high fever with diarrhea, 2; high fever with rhinopharyngitis, 1; rhinopharyngitis with conjunctivitis and otitis, 1. The sequences of the seven amplified products were similar for all cases and had 99% homology with the previously described N gene sequence. PMID- 12553304 TI - Case report of Staphylococcus aureus endocarditis after navel piercing. AB - A 13-year-old girl with surgically corrected congenital heart disease presented with a 3-day history of fever 1 month after piercing her navel. An echocardiogram demonstrated a vegetation within her right ventricle to pulmonary artery conduit, and several blood cultures were repeatedly positive for Staphylococcus aureus. Surgical replacement of the conduit in conjunction with intravenous antibiotic therapy was curative. PMID- 12553305 TI - Varicella-zoster virus: an overlooked cause of aseptic meningitis. AB - Varicella-zoster virus causes varicella (chickenpox) and zoster (shingles). Neurologic manifestations occur in both illnesses. We describe a previously healthy child who had aseptic meningitis without exanthem caused by reactivation of varicella-zoster virus. This has not been previously reported in the pediatric literature. PMID- 12553306 TI - Human immunodeficiency virus type 1 RNA polymerase chain reaction reasonably excludes infection in exposed infants. AB - A nucleic acid sequence-based amplification (NASBA) assay that detects HIV RNA may be helpful in excluding perinatal HIV infection. We reviewed the records of 190 infants born to HIV-infected mothers. The sensitivity and specificity of the NASBA assay were 100% when measured at two time periods in the first 9 months of life. The HIV RNA NASBA assay should be considered a valid diagnostic test in this population. PMID- 12553307 TI - Ankylosis of the temporomandibular joint as a sequela of septic arthritis and neonatal sepsis. AB - Ankylosis of the temporomandibular joint (TMJ) can be a result of trauma, degenerative changes and infection or of a space-occupying lesion and, when occurring during early childhood, can result in severe functional disability and facial distortion. Septic arthritis of the TMJ is extremely rare in young infants. We report two children with TMJ ankylosis that is believed to have been a consequence of undiagnosed septic arthritis of the TMJ in the neonatal period. PMID- 12553308 TI - Abstracts from the XIVth Meeting on ADP-Ribosylation Processes. Urbino, Italy, September 24-25, 2001. PMID- 12553310 TI - Krieg Cortical Kudos 2002. The Cortical Explorer Awards. PMID- 12553312 TI - Krieg Cortical Kudos 2002. The Cortical Scholar Award. PMID- 12553314 TI - Cardio-Nephrology. Cardio Renal Anemia Syndrome. Proceedings of an International Symposium. Berlin, Germany, October 27, 2001. PMID- 12553313 TI - The Public's Health and the Law in the 21st Century: A Partnership Conference on Public Health Law. June 17-19, 2002. PMID- 12553315 TI - Abstracts of the 8th World Congress on Intensive Care and Critical Care Medicine. October 28-November 1, 2001. Sydney, Australia. PMID- 12553316 TI - Pressure ulcers: prevention should make prediction a thing of the past. PMID- 12553317 TI - Proceedings of the Yale University Workshop on Irritable Bowel Syndrome. September 26, 2001. PMID- 12553318 TI - [Will AIDS spread to the entire population of Africa south of the Sahara?]. PMID- 12553319 TI - Cover legend: Vincent T. De Vita Jr.; a member of the Editorial Academy of The International Journal of Oncology. PMID- 12553320 TI - The intergenerational transmission of fertility in contemporary Denmark: the effects of number of siblings (full and half), birth order, and whether male or female. AB - Using the Danish Fertility Database, we investigate intergenerational fertility transmission, including the relationship between the number of children born to those aged 25 and 26 years in 1994 and the number of their full sibs and half sibs. We find that the fertility behaviour of parents and their children is positively correlated, and that half-sibs and full sibs have broadly similar effects. We do not find, in this complete national population, the strong birth order effects reported in some earlier studies. Nor do we find evidence of a weakening of intergenerational fertility transmission over time, perhaps because the greater flexibility of lifestyles in this post-transitional phase provides the extended social space within which intergenerational continuities can manifest themselves. We show that members of large families are over-represented in subsequent generations - that they have far more kin than those from smaller families - and that intergenerational continuities in fertility behaviour play a substantial role in keeping fertility higher than it would be in the absence of such transmission. PMID- 12553321 TI - [Vaccinations of Adults: whom to vaccinate, and how. Proceedings of an international congress. Gibraltar, April 13-16, 2002]. PMID- 12553322 TI - Study validates quick blood test for heart failure. PMID- 12553323 TI - Landmark hormone replacement therapy study stopped after finding more harms than benefits. PMID- 12553324 TI - Bladder-sparing procedure as good as cystectomy for treating invasive bladder cancer. PMID- 12553325 TI - Heart attack and stroke prevention guidelines emphasize lifestyle factors. PMID- 12553326 TI - A new estimate of Ukrainian population losses during the crises of the 1930s and 1940s. AB - Ukraine experienced two very acute demographic crises during the Soviet era: the 1933 famine and the Second World War. While different estimates of total losses have been produced previously, we have tried here to distinguish the specific impact of the crises on mortality from their impact on fertility and migration. Taking into account all existing sources of registered data and estimates, a painstaking reconstruction of annual demographic changes has been produced and complete annual life tables have been computed for the years 1926-59. Life expectancy at birth fell to a level as low as 10 years for females and 7 for males in 1933 and plateaued around 25 for females and 15 for males in the period 1941-44. PMID- 12553327 TI - Changes in functional limitation and survival among older Taiwanese, 1993, 1996, and 1999. AB - Using data from the Survey of Health and Living Status of the Elderly in Taiwan, we investigate changes in difficulties in walking and climbing stairs, tasks that represent basic lower-body movements less likely to be influenced by changes in environment and social roles than are activities and instrumental activities of daily living. Results are shown for unadjusted prevalence rates and rates adjusted for changes in population composition. The findings indicate that Taiwan does not appear to be experiencing the improvements in functioning witnessed recently in the United States. Prevalence of functional limitation increased between 1993 and 1996 and between 1996 and 1999. One possible reason is the change in old-age survival, which appears to have benefited those who have functional limitations, especially in a severe form. The Universal Health Insurance programme, established in 1995, may have increased access to care and thus the survival of those in poorest health. PMID- 12553328 TI - Religious differentials in infant and child mortality in Holland, 1855-1912. AB - At least three kinds of hypothesis may be invoked to interpret religious differentials in mortality. They are (i) hypotheses that refer to characteristics, (ii) those that refer to lifestyle, and (iii) those that refer to the social isolation of minorities. This paper tests all three kinds of hypothesis using data on urban child mortality from the Hague just before and during the demographic transition. A hazard analysis suggests that economic and demographic characteristics do not account for much of the variation by religion. An analysis of seasonal mortality suggests that some of the variation may be explained by differences in lifestyle. The third kind of hypothesis is presented here for the first time. We suggest that the social isolation of small religious groups lowered their exposure to certain kinds of infectious disease. We use a simulation study to show that this hypothesis could account for part of the variation. PMID- 12553329 TI - Sexual risk-taking in the slums of Nairobi, Kenya, 1993-8. AB - Relatively less attention has been paid to reproductive health problems facing deprived urban residents than to those facing rural residents in the sub-Saharan Africa. This is probably because the majority of Africans live in rural areas, where they are presumed to have poorer medical, educational, and other social services. Yet, the unprecedented rate of urbanization and the accompanying disproportionate growth in the proportion of poor city residents pose new challenges for health care in the region. This study examines differences in sexual behaviour between slum residents and non-slum residents in Nairobi city. The results show that slum residents start sexual intercourse at earlier ages, have more sexual partners, and are less likely than other city residents to know of or adopt preventive measures against contracting HIV/AIDS. The findings highlight the need to treat slum residents as a sub-population uniquely vulnerable to reproductive health problems and to expend more resources in slum settings. PMID- 12553330 TI - Applying Lee-Carter under conditions of variable mortality decline. AB - The Lee-Carter method of mortality forecasting assumes an invariant age component and most applications have adopted a linear time component. The use of the method with Australian data is compromised by significant departures from linearity in the time component and changes over time in the age component. We modify the method to adjust the time component to reproduce the age distribution of deaths, rather than total deaths, and to determine the optimal fitting period in order to address non-linearity in the time component. In the Australian case the modification has the added advantage that the assumption of invariance is better met. For Australian data, the modifications result in higher forecast life expectancy than the original Lee-Carter method and official projections, and a 50 per cent reduction in forecast error. The model is also expanded to take account of age-time interactions by incorporating additional terms, but these are not readily incorporated into forecasts. PMID- 12553331 TI - Work-related acute leukemia and mucor mycosis in a boat-builder. PMID- 12553332 TI - Combination of nonmyeloablative stem cell transplantation and Imatinib in accelerated phase CML. PMID- 12553333 TI - Basiliximab for the treatment of graft rejection in haploidentical peripheral blood stem cell transplantation. PMID- 12553334 TI - Life-threatening complication of Rituximab in a child. PMID- 12553335 TI - American natural. [Review of: Chaplin, J. Subject matter: technology, the body, and science on the Anglo-American frontier, 1500-1676. Cambridge, Mass: Harvard University Press, 2001]. PMID- 12553336 TI - Tourism and the market revolution. [Review of: Lewis, CM. Ladies and gentlemen on display: planter society at the Virginia Springs, 1790-1860. Charlottesville: University Press of Virginia, 2001; Corbett, T. The Making of American resorts: Saratoga Springs, Ballston Spa, Lake George; Sterngass, J. First resorts: pursuing pleasure at Saratoga Springs, Newport, and Coney Island. Baltiremore: Johns Hopkins University Press, 2001]. PMID- 12553337 TI - Our corpses, our selves: anatomy and identity in American society. [Review of: Sappol, M. A traffic of dead bodies: anatomy and embodied social identity in nineteenth-century America. Princeton: Princeton University Press, 2002]. PMID- 12553338 TI - Do good fences make good neighbors? [Review of: LaDow, B. The medicine line: life and death on a North American borderland. New York: Routledge Press, 2001]. PMID- 12553339 TI - Teaching the illusion of facilitated communication. PMID- 12553340 TI - CT portography by multidetector helical CT: comparison of three rendering models. AB - PURPOSE: To assess the value of multidetector CT portography in visualizing varices and portosystemic collaterals in comparison with conventional portography, and to compare the visualizations obtained by three rendering models (volume rendering, VR; minimum intensity projection, MIP; and shaded surface display, SSD). METHODS: A total of 46 patients with portal hypertension were examined by CT and conventional portography for evaluation of portosystemic collaterals. CT portography was performed by multidetector CT (MD-CT) scanner with a slice thickness of 2.5 mm and table feed of 7.5 mm. Three types of CT portographic models were generated and compared with transarterial portography. RESULTS: Among the 46 patients, 48 collaterals were identified on CT transverse images, while 38 collaterals were detected on transarterial portography. Forty four of 48 collaterals identified on CT transverse images were visualized with the MIP model, while 34 and 29 collaterals were visualized by the VR and SSD methods, respectively. The average CT value for the portal vein and varices was 198 HU with data acquisition of 50 sec after contrast material injection. CONCLUSIONS: CT portography by multidetector CT provides excellent images in the visualization of portosystemic collaterals. The images of collaterals produced by MD-CT are superior to those of transarterial portography. Among the three rendering techniques, MIP provides the best visualization of portosystemic collaterals. PMID- 12553341 TI - Interobserver variation in diagnosis of dementia by brain perfusion SPECT. AB - Brain perfusion SPECT (BP-SPECT) has characteristic patterns of abnormality, enabling the differential diagnosis of dementia. The purpose of this study was to measure interobserver variations in the diagnosis of dementia using BP-SPECT. BP SPECT images of 57 cases, 19 of Alzheimer's disease (AD), eight of multi-infarct dementia (MID), three of Pick's disease, five of other dementias, and 22 normal controls, were interpreted by ten nuclear medicine physicians with varying levels of experience. Brain MR images of the cases were then interpreted apart from SPECT. The physicians independently rated all of the diagnoses listed beforehand according to a five-point scale, with clinical information provided. Receiver operating characteristic (ROC) curves and the area under the ROC curve (Az) were calculated. Az varied from 0.48 to 0.87. Mean Az's were significantly larger (p<0.05) in the diagnosis by SPECT than in that by MRI (0.715 and 0.629 for dementia vs. normal, 0.670 and 0.560 for AD or MID vs. normal, 0.610 and 0.416 for AD vs. normal, and 0.672 and 0.412 for AD vs. MID, respectively). Considerable interobserver variation was present in BP-SPECT interpretation. BP SPECT may be more effective for the evaluation of dementia than MRI when the same nuclear medicine physicians interpret both images. PMID- 12553342 TI - Endovascular treatment of anterior cerebral artery aneurysms using Guglielmi detachable coils: mid-term clinical evaluation. AB - PURPOSE: Clinical and angiographic evaluations were performed in 19 consecutive patients with anterior communicating artery (ACoA) aneurysms treated by Guglielmi detachable coil (GDC) embolization between March 1997 and April 1999. METHODS: The angiographic criteria for endovascular treatment were 1) small (< 15 mm) aneurysm with a neck smaller than the fundus and 2) distinguishable aneurysm neck and relationship to the parent artery. Initially, we also included patients with anticipated surgical difficulties. These criteria were later expanded to include patients with conditions amenable to surgical clipping. Eleven patients presented with subarachnoid hemorrhage (SAH), and eight patients were asymptomatic. The most recent clinical follow-up was obtained by telephone interview with the patients and with their close relatives, to examine post-treatment personality changes. RESULTS: Complete obliteration (68%) or near-complete obliteration (neck remnant) (32%) of the ACoA aneurysm was achieved in all patients. Six-month follow-up angiography performed in 15 patients demonstrated no recurrence of aneurysms. The clinical outcome was good recovery in 15 (80%), moderately disabled in one (5%), vegetative survival in one (5%), and death in two (10%). The patients who had good recovery did not demonstarate significant personality or behavioral change. No late rebleeding had occurred at the time of writing, a minimum of 33 months postoperatively. CONCLUSION: The outcome of endovascular treatment of ACoA aneurysms was satisfactory, especially in post-treatment psychosocial sequelae. PMID- 12553343 TI - CyberKnife stereotactic irradiation for metastatic brain tumors. AB - BACKGROUND: The CyberKnife provides a new technique for performing frameless stereotactic irradiation. So far, few reports have been published on clinical outcomes obtained with the CyberKnife. This report summarizes our clinical experience with CyberKnife irradiation for metastatic brain tumors. MATERIALS AND METHODS: Seventy-seven lesions (48 patients) were evaluated and analyzed, and 66 lesions in 41 patients were treated with stereotactic radiosurgery (SRS). The prescribed dose was 9 to 30 Gy. RESULTS: Freedom from progression of the tumors was more likely with a prescribed dose of at least 24 Gy than with one of less than 20 Gy (p=0.0244; log-rank test). The CR (complete response) rate was significantly higher when D99 was at least 24 Gy (p=0.0045). There were no severe side effects. CONCLUSION: Stereotactic irradiation with the CyberKnife for metastatic brain tumors is effective and safe. D99 should be at least 24 Gy for CyberKnife SRS treatment. PMID- 12553344 TI - Monte Carlo calculation of depth doses for small field of CyberKnife. AB - PURPOSE: A Monte Carlo (MC) model of CyberKnife was developed as a quality assurance tool. The percentage depth dose (%dd) was verified by using this model. MATERIALS AND METHODS: An MC model was developed with Electron Gamma Shower version 4 (EGS4) in two steps: (1) a model of the CyberKnife treatment head and (2) a model of the collimator and phantom. The bremsstrahlung spectrum was calculated using the first model, and this spectrum was then used to calculate %dds with the second model. The calculated %dds for a large field (60 mm diameter) and three small fields (30, 15, and 5 mm diameter) were compared with those measured with a diamond detector. RESULTS AND DISCUSSION: The MC-calculated and measured %dd-curves for the 60 mm diameter field were in excellent agreement (<1.85%), thus confirming the validity of the model. Discrepancies between the calculated and measured %dd-curves increased with decreasing field size, with considerable discrepancy (11.62%) for the 5 mm diameter field due to lateral electron disequilibrium. Accurate dose can be determined with MC even in small fields. CONCLUSION: The MC technique can provide reliable standard data for accurate dose delivery with high-technology radiotherapies using small beams. PMID- 12553345 TI - Role of radiation therapy in the management of primary mediastinal large B-cell lymphoma (PMLBL). AB - PURPOSE: PMLBL is a rare disease in Japan, and its optimal management and prognosis remain to be examined. A retrospective analysis of combined modality treatment in PMLBL was performed. MATERIALS AND METHODS: Three women and four men (mean age, 36 years) were found to have PMLBL. Six patients had stage II disease, and one patient stage III disease. Each patient had a mediastinal tumor over 9 cm in diameter. The numbers of risk factors according to the international prognostic index (IPI) and modified tumor score (mTS) were 2 and 2 or 3, respectively. All patients were treated by doxorubicin-based chemotherapy. Two patients underwent tumor resection. Radiation therapy of 27.3 to 40 Gy (mean, 32 Gy) was delivered, after chemotherapy in six patients, and before chemotherapy in one. RESULTS: Only one stage IIE patient recurred in the bilateral kidneys and had a fatal outcome. The remaining six patients remain disease-free at follow-up ranging from seven to 126 months. Positive accumulation of gallium scintigraphy after chemotherapy was converted to negative by radiation therapy. CONCLUSIONS: The favorable prognosis of PMLBL was confirmed in this study. Radiation therapy should preferably be delivered to all patients with PMLBL after chemotherapy. PMID- 12553346 TI - Referral to radiologists: the value of expertise and specialty training in the practice of radiology (more than a doc in a dark box). PMID- 12553347 TI - Availability, requesting practices, and barriers to referral for high-resolution CT of the lungs: results of a survey of U.S. pulmonologists. AB - RATIONALE AND OBJECTIVES: The authors performed this study to assess the availability of high-resolution computed tomography (CT), current referral practices, and potential barriers to the clinical use of high-resolution CT in patients with diffuse lung disease. MATERIALS AND METHODS: The authors sent a survey to 450 pulmonologists who were members of the American College of Chest Physicians. They sought information about the availability of high-resolution CT services, explored the physicians' current requesting practices and the monthly number of referrals, and assessed the influence of six factors on the decision to refer. RESULTS: The authors received completed surveys from 230 pulmonologists (52.6%). High-resolution CT services were available to all respondents and had been used by 226 (98.3%) of them during the preceding 12 months. Two-thirds of respondents referred one to four patients per month, and one-third referred five or more. Monthly referral volume did not significantly vary among different practice types. The pulmonologists expressed a preference for hospital-based imaging facilities and routinely reviewed the images from the examinations they ordered. The most frequent reason for high-resolution CT was classification of a known interstitial process. High-resolution CT was infrequently requested to evaluate symptomatic immunocompromised patients. Confidence in the radiologist's high-resolution CT interpretation was the most important factor influencing the referral decision, and radiation dose to the patient was the least important. CONCLUSION: High-resolution CT services are widely available to and frequently requested by U.S. pulmonologists in a variety of practice settings. High resolution CT may be underutilized in symptomatic immunocompromised patients. Radiologists should be aware that the perception of their skill in high resolution CT interpretation is an important determinant in the pulmonologist's decision to refer. PMID- 12553348 TI - Comparison of full-field digital mammography to screen-film mammography with respect to diagnostic accuracy of lesion characterization in breast tissue biopsy specimens. AB - RATIONALE AND OBJECTIVES: This study was performed to investigate whether full field digital mammography (FFDM) is at least as accurate as screen-film mammography with respect to breast lesion characterization. MATERIALS AND METHODS: Seventy-nine breast surgical specimens were obtained by means of preoperative needle localization with surgical excision from 79 patients. The specimens were imaged with both screen-film mammography and FFDM. Six radiologists specialized in breast imaging analyzed both sets of images and characterized the visualized lesions on a five-point scale: 1, definitely not malignant; 2, probably not malignant; 3, possibly malignant; 4, probably malignant; and 5, definitely malignant. Receiver operating characteristic curve analysis of the data was then performed to assess for differences between modalities in the radiologists' ability to predict breast malignancy. RESULTS: The areas under the receiver operating characteristic curves for the prediction of breast malignancy in surgical biopsy specimens were not statistically significantly different for FFDM and screen-film mammography. CONCLUSION: The results demonstrate that with breast surgical specimens, FFDM is similar in diagnostic accuracy to screen-film mammography. PMID- 12553349 TI - MR relative cerebral blood flow mapping of Alzheimer disease: correlation with Tc 99m HMPAO SPECT. AB - RATIONALE AND OBJECTIVES: To evaluate the value of magnetic resonance (MR) perfusion imaging for diagnosis of Alzheimer disease (AD), the authors compared relative cerebral blood flow (CBF) maps obtained with MR perfusion imaging and technetium-99m hexamethyl-propyleneamine oxime (HMPAO) single photon emission computed tomography (SPECT) in patients with AD. MATERIALS AND METHODS: Eight patients with AD were studied with MR perfusion imaging and HMPAO SPECT. The relative CBF maps from the two techniques were spatially coregistered, and relative CBF values in 13 cerebral gray matter regions (total, 26 regions of interest) were compared with regression analysis. To evaluate the degree of deviation of each brain region from the regression line, a P value for the residual was calculated for each region. RESULTS: A significant overall correlation was seen between the relative CBF values produced by the two techniques (r = .68, P < .0001). Smaller P values for the residuals were obtained in the anterior cingulate cortex (P = .05) and posterior cingulate cortex (P < .001), indicating larger deviations in these regions. When data from these two regions were eliminated, the correlation coefficient rose to 0.80 (P < .0001). CONCLUSION: Despite fairly large discrepancies in the anterior and posterior cingulate cortices, the relative CBF map obtained with MR imaging is generally in close agreement with the HMPAO SPECT map, suggesting that MR perfusion imaging can provide clinically useful information regarding CBF abnormalities in patients with AD. PMID- 12553350 TI - Proteoglycan depletion-induced changes in transverse relaxation maps of cartilage: comparison of T2 and T1rho. AB - RATIONALE AND OBJECTIVES: The authors performed this study to (a) measure changes in T2 relaxation rates, signal-to-noise ratio (SNR), and contrast with sequential depletion of proteoglycan in cartilage; (b) determine whether there is a relationship between the T2 relaxation rate and proteoglycan in cartilage; and (c) compare the T2 mapping method with the spin-lattice relaxation time in the rotating frame (T1rho) mapping method in the quantification of proteoglycan induced changes. MATERIALS AND METHODS: T2- and T1rho-weighted magnetic resonance (MR) images were obtained in five bovine patellae. All images were obtained with a 4-T whole-body MR unit and a 10-cm-diameter transmit-receive quadrature birdcage coil tuned to 170 MHz. T2 and T1rho maps were computed. RESULTS: The SNR and contrast on the T2-weighted images were, on average, about 43% lower than those on the corresponding T1rho-weighted images. The T2 relaxation rates varied randomly without any particular trend, which yielded a poor correlation with sequential depletion of proteoglycan (R2 = 0.008, P < .70). There was excellent linear correlation between the percentage of proteoglycan in the tissue and the T1rho relaxation rate (R2 = 0.85, P < .0001). CONCLUSION: T2-weighted imaging neither yields quantitative information about the changes in proteoglycan distribution in cartilage nor can be used for longitudinal studies to quantify proteoglycan-induced changes. T1rho-weighted imaging, however, is sensitive to sequential depletion of proteoglycan in bovine cartilage and can be used to quantify proteoglycan-induced changes. PMID- 12553351 TI - Local differences in the trabecular bone structure of the proximal femur depicted with high-spatial-resolution MR imaging and multisection CT. AB - RATIONALE AND OBJECTIVES: The authors performed this study to investigate structural variations in the trabecular bone of the proximal femur at high resolution magnetic resonance (MR) imaging and high-resolution multisection computed tomography (CT). MATERIALS AND METHODS: Bone mineral density (BMD) was measured in 36 proximal human femur specimens by using dual x-ray absorptiometry. High-resolution MR imaging was performed at 1.5 T with an in-plane spatial resolution of 0.195 x 0.195 mm and a section thickness of 0.3 and 0.9 mm. Multisection CT was performed with an ultra-high-resolution protocol; images were obtained with an in-plane spatial resolution of 0.25 mm and a section thickness of 1 mm. In a subset of these specimens, micro CT was performed with an isotropic spatial resolution of 30 microm. Identical regions of interest (ROIs) were used to analyze images obtained with MR imaging, multisection CT, and micro CT. Trabecular bone structural parameters were obtained, and the parameters from the individual imaging modalities and BMD were correlated. RESULTS: Significant differences concerning the trabecular microarchitecture between the individual ROIs were demonstrated with multisection CT and MR imaging. A number of the correlations between structural parameters derived with multisection CT, MR imaging, micro CT, and BMD measurements were significant. For MR imaging, threshold technique and section thickness had an effect on structural parameters. CONCLUSION: Structural parameters obtained in the proximal femur with multisection CT and high-resolution MR imaging show regional differences. These techniques may be useful for depicting the trabecular architecture in the diagnosis of osteoporosis. PMID- 12553352 TI - Time-resolved three-dimensional pulmonary MR angiography and perfusion imaging with ultrashort repetition time. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to implement ultrafast, multiphase three-dimensional (3D) magnetic resonance (MR) angiography and perfusion imaging after bolus injection of contrast medium to generate preliminary validation of parameters in a pig model and to illustrate potential applications in patients with lung abnormalities. MATERIALS AND METHODS: Five healthy volunteers, five patients, and three pigs underwent rapid, time-resolved pulmonary MR angiography and perfusion imaging on a 1.5-T MR imager. All patients had undergone correlative computed tomographic or conventional angiography. The pulse sequence was a 3D spin-warp, gradient-echo acquisition with a repetition time of 1.6 msec and an echo time of 0.6 msec. Each 3D acquisition lasted 2-3 seconds, and 8-16 sequential measurements were made in each study. Artificial pulmonary emboli were generated in pigs with gelatin sponge. All patients had diseases of the pulmonary circulation (as confirmed with other studies). RESULTS: Multiphasic, time-resolved pulmonary parenchymal enhancement was demonstrated in all healthy subjects and animals. All segmental (n = 100) and subsegmental (n = 200) branches were identified in the healthy subjects. Perfusion deficits were clearly demonstrated in all pigs after gelatin embolization. Perfusion defects were identified in two patients with lung disease. Abnormalities of the pulmonary vasculature were clearly identified in the patient group. CONCLUSION: Dynamic time-resolved 3D pulmonary MR angiography and perfusion imaging is feasible in humans as well as in animals. Induced perfusion deficits are identifiable after artificial embolization in pigs. Combined pulmonary MR angiography and parenchymal (perfusion) imaging may improve evaluation of the pulmonary circulation in a variety of conditions. PMID- 12553353 TI - The art of asking: teaching through questioning. PMID- 12553354 TI - Key component of resident selection: the semistructured conversation. PMID- 12553355 TI - Volume and impact of second-opinion consultations by radiologists at a tertiary care cancer center: data. AB - RATIONALE AND OBJECTIVES: Patients with cancer who are referred to a dedicated oncology center usually have undergone previous imaging studies that the oncologists typically desire to have reviewed by radiologists. Such reinterpretations can be complex and time-consuming, yet many institutions do not systematically account for them as part of the total workload. The purpose of this study was to ascertain the numbers and types of second-opinion consultations performed by radiologists at a tertiary care cancer center, and to assess their effect on work volume. MATERIALS AND METHODS: A survey of referring clinicians was undertaken to evaluate the numbers and types of second-opinion consultations requested of radiologists at the Dana Farber Cancer Institute during a 12-month period. Consultations included review of studies from outside institutions, and cases from Dana Farber in which further comparison was needed. The number of consultations requiring additional tumor size measurements was tallied. The mean daily number of new studies interpreted by radiologists was used as a benchmark of work volume. RESULTS: Radiologists performed 4,664 consultations during 254 workdays, interpreting a mean of 18 additional studies (range, 4-42) per day as a result of referrals for second opinion. These included 3,638 (78%) cross sectional studies (ie, computed tomographic [CT], magnetic resonance [MR], and ultrasound [US] studies), 674 (14%) mammograms, 220 (5%) plain radiographs, 132 (3%) nuclear medicine scans, and one galactogram. Of the 4,664 consultations, 1,306 (28%) were performed to obtain tumor measurements, many of these involving five to 10 bidimensional calculations per study. A mean of 101 new examinations per day was performed by radiologists during the same 12-month period, including cross-sectional studies (CT and US scans) (56%), plain radiographs (34%), and mammograms (11%). MR imaging was not performed. CONCLUSION: Second-opinion consultations increased the average daily work volume by 18%. This has implications for workforce, as well as for compensation in terms of relative value units and finances for this previously unquantified service. PMID- 12553357 TI - Volunteers. PMID- 12553356 TI - Learning to report. PMID- 12553358 TI - Our family in radiology. PMID- 12553359 TI - Educational reform: problem-based learning at Chang Gung University, an overview. AB - Problem-based learning is an educational method characterized by utilization of problems involving patients as a context for students to learn problem-solving skills and acquire knowledge about basic and clinical science. It is a core curriculum in the new pathway of educational reform. Chang Gung University and Chang Gung Memorial Hospital began a program of educational reform in 1998. However, many difficulties were experienced at the very beginning including the capabilities of students, manpower resources, and a transition period of implementation. After a series of communication, education, practice, and system reforms, the problem-based learning curriculum is now performing quite smoothly. This paper reviews the history of and our experiences with educational reform including preparation, implementation, the content and range of reform, accompanying package, committee establishment, tutor training, assessment of tutors and students, etc. The fact that our education reform began from clinical aspects and then progressed to basic medicine is unique, and a hybrid curriculum with a model of parallel progress in traditional and new curricula was created using internal medicine as a reference for other departments. Finally, we hope that the effect of well-designed reforms in facilitating learning should not be underestimated. PMID- 12553360 TI - Overexpression of damaged-DNA-binding protein 2 (DDB2) potentiates UV resistance in hamster V79 cells. AB - BACKGROUND: Ultraviolet radiation (UV) damaged-DNA binding (DDB) activity comprises two major components: damaged-DNA binding protein 1 (DDB1) and 2 (DDB2). Although the function of DDB is unclear, mutation on DDB2 is associated with cellular sensitivity to a variety of genotoxic agents including UV. It has been suggested that DDB2 may play a role in UV-induced DNA repair. However, evidence that DDB2 involves in DNA repair and UV sensitivity is lacking. METHODS: To examine the role of DDB2, we established DDB2-overexpressing hamster V79 cell lines, V79ddb2, by stable transfection with full-length open reading frame of human ddb2 cDNA. Cells were irradiated with UV and determined its DNA repair activity by testing the remaining photoproducts on the chromatin and measuring the plasmid reactivation, respectively. UV induced cytotoxicity was determined by the colorimetric assay (MTT assay), and apoptotic cells exhibiting morphological features of chromatin condensation and nuclear fragmentation were counted after 4 diamidino-2-phenylindole (DAPI) staining. RESULTS: DDB activity was increased in DDB2-overexpressing cell lines. Analysis on DNA repair indicated that UV photoproducts were removed in a time-dependent manner and there was greater than 50% of damage removed within 12 h in DDB2-overexpressing cells. In contrast, nearly all the damage remained unrepaired in V79 cells. However, using bacterial CAT gene as a reporter, both V79 and V79ddb2 cells demonstrated no difference in the reactivation of plasmid DNA carrying UV damage. These results suggest that DDB2 may involve in repair of bulky genomic DNA damage. Although a maximum of only 30% of apoptosis was induced, UV irradiation caused a dose-dependent apoptosis and cytotoxicity in these cell lines. V79ddb2 cells displayed resistance to UV-induced apoptosis and cytotoxicity. CONCLUSION: Our findings indicate that overexpression of DDB2 in V79 cell potentiates DNA repair and protects cells from UV-induced cytotoxicity. These results also suggest that DDB2 may be involved in the development of UV resistance. PMID- 12553361 TI - Quantitative analysis of chimerism after allogeneic peripheral blood stem cell transplantation. AB - BACKGROUND: For peripheral blood stem cell transplantation (PSCT), several engraftment analysis methods have been performed including detection of restriction fragment length polymorphisms and amplification of polymorphic genetic loci. To facilitate monitoring of the engraftment, a quantitative, non isotopic method using a short tandem repeat (STR) marker has been set up in our laboratory. METHODS: DNAs from pretransplant recipients and donors were amplified with the AmpFISTR Profiler Plus kit that contains 9 STR markers. The fluorescent polymerase chain reaction products were then fractionated on polyacrylamide gels in an ABI PRISM 377 DNA sequencer. Results were analyzed using GeneScan 2.1 software. We selected the best markers as informative alleles which can distinguish donor from recipient. For quantitative analysis of the engraftment, we prepared a mixed chimeric sample by mixing pretransplant recipient and donor DNAs in different ratios to produce a standard curve. After amplifying the posttransplant recipient DNA, we were able to detect the extent of engraftment by interpolating the percent peak area of the informative alleles from this standard curve. RESULTS: We retrospectively analyzed 10 patients who had received allogeneic PSCT. Two of them showed some degree of mixed chimerism indicating leukemic relapse. In case one, 38.7% of the recipient DNA was first detected in the third month after PSCT. In case two, 6.5% of the recipient DNA was first detected in the tenth month after PSCT. CONCLUSION: In summary, this method provides an accurate, quantitative, and early assessment of mixed chimerism in posttransplant patients. Such information may be useful to guide implementation of additional treatment to circumvent graft failure or relapse in the future. PMID- 12553362 TI - Clinical analysis of 1048 children with developmental delay. AB - BACKGROUND: Children with developmental delay (DD) have a variety of problems in developmental functions. The purposes of this study were to analyze the underlying diseases and risk factors in children with different functional delays. METHODS: We collected data on 1048 children who underwent assessments of developmental function, related diseases, and risk factors. All children were classified into 6 functional delay groups: cognitive, speech, motor, pervasive, global, and non-specific DDs. Differences in related diseases and risk factors of the 6 functional delay groups were determined. RESULTS: Most children had global (51.2%), speech (21.9%), and motor (13.9%) delays. Approximately 62.8% of children were associated with biological factors (19% with genetic defects or congenital anomalies, 16.5% with central nervous system lesions, 13.9% with prematurity/low birth body weight, and 13.4% with neonatal insult). We could not identify the risk factors in 36.6% of the children. Most children with motor delay had brain/neuromuscular diseases and were associated with risks of prematurity or low birth body weight; while most children with global delay had brain neuromuscular diseases or psychological/mental disorders and were associated with risks of genetic defects or congenital anomalies. CONCLUSION: Our findings suggest that there are heterogeneous risk factors and related diseases in children with different kinds of functional delay. PMID- 12553363 TI - Ultrasound-guided transvaginal cyst aspiration for the management of pelvic pseudocyst: a preliminary experience. AB - BACKGROUND: While some research has been reported on the use of transvaginal ultrasound-guided aspiration of ovarian cysts, none has been reported for pseudocysts. Sometimes laparotomy patients experience recurrent pelvic psuedocysts for which they may need to undergo another laparotomy. The use of transvaginal ultrasound-guided aspiration of pelvic pseudocysts may make repeated laparotmies unnecessary. METHODS: Between April 1993 and October 2001, 14 outpatients with postoperative pelvic pseudocysts underwent transvaginal ultrasound-guided aspirations with or without surgical starch irrigation. All patients had previously undergone pelvic surgeries for adenomyosis and leiomyoma, except one was stage lb cervical cancer. A total of 25 treatment cycles by transvaginal ultrasound-guided aspiration were included: 15 cycles with starch and 10 cycles without. RESULTS: The 13 patients with more than 6 months follow-up had a total of 25 aspiration cycles, 22 of which (88%) had no recurrent cyst by the end of their 6-month follow-up period. Eighteen out of 25 cycles (72%) had no evidence of recurrence after 12 months of follow-up. Five patients needed only one aspiration to become disease free and six patients needed two aspirations. Only 2 patients needed 3 or more aspirations procedures. After 12 months of follow up, 33% of those irrigated with starch had a recurrent pelvic pseudocyst while 30% without starch experienced recurrence. CONCLUSION: Because it eliminates necessity of repeat laparotomy, transvaginal ultrasound-guided aspiration of pelvic pseudocysts may become a preferred treatment for recurrent pelvic pseudocysts. PMID- 12553364 TI - Evaluation of problem-based learning education after clerkship at the Chang Gung University School of Medicine. AB - BACKGROUND: The Chang Gung University School of Medicine adopted problem-based learning (PBL) education 3 years ago. A questionnaire was designed to evaluate the effectiveness of this teaching method, and the results were analyzed to determine statistical significance. METHODS: In June 2001, all the interns in the Medical and Surgical departments of the Chang Gung Memorial Hospital were compulsorily assessed using a newly developed questionnaire, which was provided to the residents, chief resident, and attending doctors. The questions involved the interns' ability to perform 10 essential skills, namely (1) problem searching, (2) problem solving, (3) initiative learning, (4) thinking process, (5) establishing the patient-doctor relationship, (6) establishing the doctor nurse relationship, (7) interaction with peers, (8) professional knowledge, (9) clinical techniques, and (10) medical notes writing. Forty-three completed questionnaires, evaluating 25 interns, were returned. Of these 25 individuals, 14 had participated in PBL education and 11 had been taught using the conventional variant. RESULTS: No statistically significant differences were demonstrated for gender or average school records between the interns who had been taught using the PBL and conventional methods. Statistically significant superiority was demonstrated for interns educated using PBL in three of 10 areas including, thinking process, professional knowledge, and clinical techniques. CONCLUSION: Analysis of the questionnaire results clearly demonstrated that the introduction of the PBL method of teaching at the university was efficacious in terms of the competence demonstrated by the interns when entering clinical practice. PMID- 12553365 TI - Primary malignant melanoma of the tongue. AB - The oral cavity is a rare location for the development of primary malignant melanoma. The most common primary lesion sites are the palate and gingiva. Melanoma of the tongue is specifically uncommon. A 66-year-old woman was referred to our clinic with a complaint of a huge, painless, black, discolored mass on the right side of the oral tongue for 7 years. There were no cutaneous lesions suggestive of malignant melanoma over the rest of her body. The biopsy of the tongue lesion revealed a histopathology consistent with primary malignant melanoma. Computed tomography of the neck showed no significant cervical lymphadenopathy. Chest radiograph, whole body bone scanning, and abdominal sonography revealed no definite distal metastatic lesions. She received composite resection of the tumor on the right side of the tongue and right functional neck dissection. The patient had an uneventful recovery and received regular follow-up examinations. She was free of disease for more than 2 years. The treatment principle for primary tongue melanoma is wide surgical excision. Early diagnosis will be promoted by careful oral examination and early biopsy of pigmented and non-pigmented masses. We reviewed the published reports in the English literature since 1970 and fewer than 30 cases of primary tongue melanoma were presented. We present a case report and a review of the relevant literature. PMID- 12553366 TI - Nasal septal perforation caused by invasive fungal sinusitis. AB - Nasal septal perforation presents a distinct challenge to otorhinolaryngologists, and is a problem for patients. Although it has a variety of causes, previous septal surgery is the most common reason. We present a 57-year-old woman who had recurrent chronic sinusitis. A left nasal mass was noted and excised via endoscopic sinus surgery. Invasive aspergillosis sinusitis was proven both grossly and histopathologically, and a nasal septal perforation was also noted during the operation. Although there has been only a single other case presented by Siberry in 1997, we postulate that perforation of the nasal septum as with the case described herein is a rare complication of invasive fungal sinusitis. PMID- 12553367 TI - Skin necrosis following a recombinant interferon-beta-1b injection. AB - Recombinant interferon beta-1b (INF-beta-1b) has been proven to be an effective means of treating relapsing-remitting multiple sclerosis (MS). Adverse reactions to interferon therapy have been well documented. The most common side effects are transient influenza-like symptoms, including fever, fatigue, nausea, and myalgia. Cutaneous necrosis has occasionally been reported, mostly involving small and limited lesions. This article describes an MS patient who developed multiple large, deep cutaneous ulcers on INF-beta-1b injection sites, which subsequently required surgical treatment. Vessel thrombosis in the subcutaneous fatty layer and the clinical appearance of livedoid erythema beside the ulcers indicated that INF-3-1b may have caused skin necrosis through its vascular effects. PMID- 12553368 TI - Endogenous Candida endophthalmitis after two consecutive procedures of suction dilatation and curettage. AB - Endogenous Candida endophthalmitis (ECE) is a rare disease. We present a patient with Candida endophthalmitis after two consecutive procedures of suction dilatation and curettage for elective abortion. A 24-year-old single woman who received a suction dilatation and curettage one week ago developed pain and blurred vision in the right eye. Endogenous Candida endophthalmitis was diagnosed and treated with oral fluconazole and pars plana vitrectomy with adjunction of intravitreal amphotericin B injection. The vitreous culture revealed Candida albicans. The vitreous inflammation subsided greatly after the initial treatment but flared up after the second dilatation and curettage for incomplete abortion 5 days after the vitrectomy. The oral fluconazole was replaced by intravenous amphotericin B, and a second vitrectomy with injection of intravitreal amphotericin B was performed. Postoperatively, the intraocular inflammation resolved gradually. Six months after the second vitrectomy, the best-corrected visual acuity in the right eye was 20/25. The excellent visual acuity of this patient was attributed to the early diagnosis and aggressive treatment. For patients with mild disease, less toxic oral fluconazole as the systemic antifungal agent instead of more toxic intravenous amphotericin B has been recommended. For those with advanced disease, intravitreal amphotericin B in conjunction with vitrectomy has been advocated by many eye surgeons. PMID- 12553369 TI - Hypoglycemia probably due to accidental intake of repaglinide. AB - This report focuses on a 16-year-old girl afflicted with hypoglycemia after administration of medications for gastrointestinal symptoms. Repaglinide-induced hypoglycemia was suspected when a tablet of repaglinide was noted in the drug package that she had been given. As the use of various types of oral hypoglycemic agents has increased, a definitive diagnosis of drug-induced hypoglycemia has become difficult. It is dangerous for a patient to take oral hypoglycemic agents without the knowledge of hypoglycemic symptoms and initial management. We present this case and review the characteristics of repaglinide to remind physicians and pharmacists to pay more attention to this situation. PMID- 12553370 TI - A critical review of vascular endothelial growth factor (VEGF) analysis in peripheral blood: is the current literature meaningful? AB - Vascular endothelial growth factor (VEGF) is a potent angiogenic growth factor with a key role in many physiological and pathological processes. Investigation into the implications of circulating levels of this cytokine is progressing at an exponential rate. However, there are important inconsistencies between reports ranging from method of sample collection, processing, software manipulation and data interpretation and controversy as to whether plasma, serum or whole blood will provide the best prognostic information. Different techniques of centrifugation and temperature on sample handling and the impact of in vitro collection of blood on subsequent VEGF results have not been fully appreciated. We provide a critical review of the literature, report the results of our further investigations, suggest a uniform protocol for handling blood samples and highlight previously unsuspected problems in data interpretation. PMID- 12553371 TI - Inhibition of tumor progression and neoangiogenesis using cyclic RGD-peptides in a chemically induced colon carcinoma in rats. AB - Integrins are cell surface molecules that mediate cell adhesion, but are also important regulators of tumor cell interactions with their microenvironment, tumor cell survival and growth. In addition, the alpha(v)beta3-integrins appear to be critical for microvessel formation in tumor-induced neoangiogenesis. The present study is the first to investigate the effects of therapeutic alpha(v)beta3-integrin inhibition in a chemically induced tumor model that largely resembles human colon carcinomas. Tumor induction was performed in 47 male Sprague-Dawley rats using 1,2 dimethylhydrazin (21 mg/kg) twice a week. After 20 weeks of tumor induction, 100% of the animals developed adenocarcinomas with a median of 13.5 macroscopic tumor nodules (range 12-17), but no distant metastases. During further tumor induction for an additional 10 weeks, rats were treated three times/week with (a) 15 mg/kg RGDfV-peptide that can block vitronectin and fibronectin receptors; (b) an equimolar amount of an ineffective cyclic control peptide; or (c) with equimolar amounts of a linear RGDS-peptide. At the end of this treatment period, rats were sacrificed, and tumor load was quantified macroscopically and confirmed by histological examination. For investigation of the involvement of tumor-induced neoangiogenesis microvessel, density was determined using CD31-immunostaining. After 30 weeks, control animals (group B) had 5-18 tumors (median 14.5). If rats were treated with RGDfV-peptide (group A), the number of tumor nodules was significantly reduced (P < 0.005) to a median of seven macroscopic tumors (range 2-10 tumors), which also represented a significant reduction (P < 0.005) compared with prior to treatment. Application of noncylic RGDS-peptides (group C) did not affect the number of tumor nodules (median 18; range 10-30 tumors). The diameters of tumor nodules were comparable (3.2-6.1 mm) in animals of all groups. In addition, microvessel density was significantly (P < 0.05) reduced in tumors in group A compared to control rats. The major side effect in the treatment group was increased susceptibility to respiratory infections. Our results demonstrate that alpha(v)beta3-integrin receptor inhibition appears to be a therapeutic strategy for colorectal cancer. In our therapeutic model, late onset of treatment with integrin-blocking peptides resulted in an inhibition of tumor growth and a reduced tumor load which appeared to be mediated, at least in part, by inhibition of neoangiogenesis. PMID- 12553372 TI - Differential assessment of angiogenic activity and of vascular survival ability (VSA) in breast cancer. AB - Recent reports provide evidence that some growth factors behave as inhibitors of the apoptosis of the endothelial cells, bringing forward the concept of vascular survival as a post-angiogenesis process. At least two different vasculature development processes occur within a tumor: the angiogenic (formation of new vessels) and the vascular survival pathway, which is devoted to the preservation of the newly-formed vessels in layers that lose contact with the adjacent normal tissue. We developed a method to assess these processes in tissue samples. We noted that differences among tumors may exist not only in the tumor angiogenic activity (TAA) but also in the vascular survival ability (VSA). One third of the highly angiogenic breast cancer cases examined had a poor ability to maintain high vessel density in inner tumor areas. Both parameters are independently related to prognosis, while VSA was directly related to tumor dimensions and node involvement. Patients with high TAA and VSA had a particularly poor prognosis. It is suggested that although cancer angiogenic activity is important for the local invasion and dissemination into vessels and lymphatics, the VSA may be important for the effective formation of viable tumor foci in lymph nodes or distant organs. Recognition and quantification of the vascular survival ability in human tumors may significantly improve the prognostic value of the assessment of tumor vasculature, and may help to stratify patients for clinical trials with novel anti-angiogenic or angiotoxic drugs. Elucidation of the pathways may provide additional targets for antiangiogenic therapy. PMID- 12553373 TI - Effects of Celebrex and Zyflo on liver metastasis and lipidperoxidation in pancreatic cancer in Syrian hamsters. AB - Selective inhibition of eicosanoid synthesis is thought to have effects on carcinogenesis in lung and colon cancer. However, it is still unknown whether pancreatic cancer might also be influenced. Therefore we evaluated the impact of selective cyclooxygenase-2 inhibitor Celebrex and selective 5-lipoxygenase inhibitor Zyflo on liver metastasis in a solid model of pancreatic adenocarcinoma in Syrian hamster. In week 33, the animals were sacrificed and incidence of pancreatic carcinomas and number and size of liver metastases were determined. Activities of antioxidative enzymes (GSHPX/SOD) and concentrations of products of lipidperoxidation were measured in liver metastases and non-metastatic hepatic tissue. The incidence (54.5 vs. 100%), number (3.17 +/- 0.98 vs. 6.75 +/- 0.71) and size (2.67 +/- 1.97 vs. 11.75 +/- 1.98 mm2) of liver metastases were decreased by combined therapy of Zyflo and Celebrex (P < 0.05). Furthermore, activities of GSHPX ([73.77 +/- 5.67]*10(5) vs. [15.49 +/- 4.02]*10(5) U/mg prot.; P < 0.05) and SOD (474.92 +/- 108.8 vs. 127.89 +/- 38.75 U/mg prot.; P < 0.05) were increased, while lipidperoxidation (0.31 +/- 0.08 nmol/mg prot. vs. 1.54 +/- 0.55 nmol/mg prot.; P < 0.05) was decreased by combination therapy, in non-metastatic hepatic tissue. Moreover, combined therapy increased lipidperoxidation in liver metastases (0.47 +/- 0.09 vs. 1.95 +/- 0.12 nmol/mg prot.; P < 0.05). Thus, a combination of Celebrex and Zyflo might be a new concept to decrease tumour growth in liver metastases in advanced pancreatic cancer. PMID- 12553374 TI - Vaccination of mice with MUC1 cDNA suppresses the development of lung metastases. AB - C57BL/6 mice were immunized intradermally with various doses of purified pCEP4 plasmid DNA containing full-length MUC1 cDNA (22 tandem repeats). Mice immunized with MUC1 DNA three times at weekly intervals had serum antibodies to a synthetic peptide corresponding to the tandem repeats of MUC1. The antibody titer correlated with the plasmid DNA dose. After the third immunization mice were injected intravenously with 5 x 10(5) 16-F10 melanoma cells that had been stably transfected with MUC1 cDNA (F10-MUC1-C8 clone cells). The number of lung metastatic nodules three weeks after inoculation of F10-MUC1-C8 cells was significantly lower in mice immunized with MUC1 plasmid DNA than in mice immunized with the vector DNA alone. Thus, the suppression of lung metastasis was antigen-specific. In vivo depletion of lymphocyte subpopulations by specific antibodies revealed that natural killer cells are the major effector cells responsible for the suppression of lung metastasis. CD4+ cells and CD8+ cells apparently played some roles too. PMID- 12553375 TI - EMMPRIN-mediated MMP regulation in tumor and endothelial cells. AB - Tumor invasion and metastasis are multistep processes which require extracellular matrix remodeling by proteolytic enzymes such as matrix metalloproteinases (MMPs). The production of these enzymes is stimulated by many soluble or cell bound factors. Among these factors, extracellular matrix metalloproteinase inducer (EMMPRIN) is known to increase in vitro stromal cell production of MMP-1, MMP-2 and MMP-3. In this study, we demonstrated that EMMPRIN-transfected MDA-MB 436 tumor cells displayed a more invasive capacity than vector-transfected cells in a modified Boyden chamber invasion assay. Using gelatin zymography and protein analyses, we showed that EMMPRIN-transfected cancer cells produced significantly more latent and active MMP-2 and MMP-3 than vector-transfected cancer cells. We found that EMMPRIN did not regulate MMP-1, MMP-9, membrane type-1 MMP (MT1-MMP) expression and had also no effect on the production of the specific tissue inhibitors of MMPs (TIMPs), TIMP-1 and TIMP-2. We also demonstrated that tumor derived EMMPRIN stimulated MMP-1, -2, and -3 without modification of MMP-9, MT1 MMP, TIMP-1 and TIMP-2 production in human umbilical vein endothelial cells (HUVEC). These data provide support for the role of EMMPRIN in tumor invasion, metastasis, and neoangiogenesis by stimulating extracellular matrix remodeling around tumor cell clusters, stroma, and blood vessels. PMID- 12553376 TI - Effect of piperine on the inhibition of lung metastasis induced B16F-10 melanoma cells in mice. AB - The effect of piperine on the inhibition of lung metastasis induced by B16F-10 melanoma cells was studied in C57BL/6 mice. Simultaneous administration of the compound with tumor induction produced a significant reduction (95.2%) in tumor nodule formation. Increased lung collagen hydroxyproline (22.37 microg/mg protein) in the metastasized lungs of the control animals compared to normal animals (0.95 microg/mg protein) was significantly reduced (2.59 microg/mg protein) in the piperine-treated animals. The high amount of uronic acid (355.83 microg/100 mg tissue) in the metastasized control animals was significantly reduced (65 microg/100 mg tissue) in the animals treated with piperine. Lung hexosamine content was also significantly reduced in the piperine-treated animals (0.98 mg/100 mg lyophilized tissue) compared to the untreated tumor-bearing animals (4.2 mg/100 mg lyophilized tissue). The elevated levels of serum sialic acid and serum gamma glutamyl transpeptidase activity in the untreated control animals was significantly reduced in the animals treated with piperine. The piperine-treated animals even survived the experiment (90 days). Histopathology of the lung tissue also correlated with the lifespan of the drug-treated animals. Our results demonstrate the antimetastatic activity of piperine, an alkaloid present in plants such as Piper nigrum and Piper longum. PMID- 12553377 TI - Modulation of proteolytic potential and differentiation by CNTF and BDNF in two mouse neuroblastoma clones: relation to invasion. AB - The effect of CNTF and BDNF on a proteolytic complement instrumental to invasion and on differentiation was studied in two murine neuroblastoma clones, N1 and N7. At the membrane level, gelatinase MMP-2--mainly the activated form--was restrained by CNTF and BDNF to a residual 34% with both factors; membrane-type 1 MMP was down-regulated to 50% (10 h) and 34% (24 h) with both factors; and urokinase-type plasminogen activator was restrained mainly by BDNF to 70%. In the medium, the two gelatinases MMP-2 and MMP-9 were mainly in zymogen form: only MMP 2 was restrained in N1 cells, while only MMP-9 was restrained in N7 cells by both factors, single or in combination. These effects were paralleled by the induction of neurite outgrowth, which was more stimulated in the less differentiated clone. These dose-dependent and transient effects make CNTF and BDNF ideal candidates for constraining the potentially invasive behavior of nervous system tumors. PMID- 12553378 TI - Role of integrin alpha(v)beta3 in the early phase of liver metastasis: PET and IVM analyses. AB - To clarify the function of integrin alpha(v)beta3 in the early stage of liver metastasis, we investigated the interactions of metastatic cells with their target organ under the actual blood flow by using positron emission tomography (PET). The cells used were CHO-K1 cells and their transfectants bearing human integrin alpha(v)beta3 cDNA (alpha(v)beta3-CHO-K1 cells). The liver accumulation of alpha(v)beta3-CHO-K1 cells was significantly higher than that of CHO-K1 cells after injection via the portal vein, whereas no significant difference was observed in the lung accumulation after tail vein injection, suggesting a specific interaction of alpha(v)beta3-CHO-K1 cells with the hepatic sinusoids. Furthermore, to clarify the precise location of each cell in the liver, i.e., to determine whether individual cells were intravascularly localized or had extravasated, we performed intravital fluorescence microscopy (IVM) on the liver by using stable transfectants bearing the green fluorescent protein (GFP) gene, namely, GFP-CHO-K1 and GFP-alpha(v)beta3-CHO-K1 cells. Both types of cells remained in the hepatic blood vessels 1 h after injection via the portal vein. On the other hand, expression of integrin alpha(v)beta3 promoted the cells to reach the extravascular region after 24 h. These results suggest the possibility that the specific accumulation of alpha(v)beta3-CHO-K1 cells in the liver is followed by migration of the cells into the extravascular region. Interestingly, the adhesion of the two types of cells to hepatic sinusoidal endothelial cells in vitro did not correspond to in vivo accumulation of these cells. Therefore, integrin alpha(v)beta3 may function to promote extravasation of integrin alpha(v)beta3-expressing tumor cells in liver through a process possibly mediated by vitronectin produced by this organ. PMID- 12553379 TI - Paracrine communication between malignant and non-malignant prostate epithelial cells in culture alters growth rate, matrix protease secretion and in vitro invasion. AB - Epithelial cancer cell invasion is facilitated by stromal cells, immune cells, endothelial cells and other epithelial cells. We have used two human papilloma immortalized prostate cell lines, CA-HPV-10 from a carcinoma and PZ-HPV-7 cells from normal prostatic epithelium to study cell-cell influences on growth, gelatinase secretion, invasion and responses to TGFbeta1. We found that co culture with CA-10 carcinoma cells stimulates proliferation of the PZ-7 epithelial line. TGFbeta1 inhibited growth of both lines, but while inhibitory effects on the CA-10 cells diminished after removal of the peptide, inhibition of PZ-7 was lasting. Interestingly, the TGFbeta-induced growth inhibition in PZ-7 cells could be partially reversed by co-culture with CA-10 cells. Co-culture with CA cells in a 3-chamber invasion assay also promoted invasion of PZ cells. CA-10 invasion was enhanced by co-culture with TGFbeta1-treated-PZ-7 cultures and this enhancement was associated with TGFbeta1-induced secretion of matrix metalloproteinase-9. Our observations suggest that interaction between prostate cancer cells and prostate epithelial cells may promote proliferation of the epithelial cell population and produce a paracrine source of MMP-9 which may facilitate early cancer cell invasion. PMID- 12553380 TI - Plasma basic fibroblast growth factor levels in colorectal cancer: a clinically useful assay? AB - Angiogenic cytokines in the plasma and serum of cancer patients may serve as 'surrogate' markers of tumour neoangiogenesis. Serum VEGF correlates with disease stage in colorectal cancer (CRC), but the role of bFGF in CRC is uncertain. This study aimed to assess plasma bFGF levels in CRC patients before treatment, during chemoradiotherapy and at one-year follow-up. Plasma samples were taken from 124 CRC patients, 26 polyp patients and 55 controls, and bFGF levels were measured by ELISA. 19 patients underwent pre-operative chemoradiotherapy. One-year follow-up samples were available from 48 disease-free patients and 18 patients with progressive disease. There were no detectable differences between plasma bFGF levels in polyp, Dukes' A or B patients (4.55, 5.77, 4.25 pg/ml, respectively), but there was a significant increase in metastatic CRC patients [Dukes' C and D (7.42 and 6.6 pg/ml; P = 0.004 and 0.048, respectively)], relative to median control levels of 4.14 pg/ml. At follow-up, there was a significant fall in plasma bFGF levels in disease-free patients (pre-op 6.09 and follow-up 3.45 pg/ml, P = 0.0004), but a non-significant rise in 18 patients with progressive disease (pre-treatment 5.90 and follow-up 9.99 pg/ml, P = 0.33). Pre-treatment plasma bFGF in patients receiving chemo-radiotherapy was similar in those with responsive and non-responsive tumours. There were no detectable changes in plasma bFGF through the adenoma-carcinoma sequence or patient groups with non-metastatic cancers. Elevated plasma bFGF was, however, associated with metastatic spread. The significant fall in bFGF in disease-free patients following therapy suggests that bFGF may be useful in clinical practice. PMID- 12553381 TI - Anaesthetic agents in adult day case surgery. AB - This study reports a review of all comparative published studies of adult day case anaesthesia in the English language up to December 2000. Ten databases were searched using appropriate keywords and data were extracted in a standardized fashion. One hundred-and-one published studies were examined. Recovery measurements were grouped as early, intermediate, late, psychomotor and adverse effects. With respect to induction of anaesthesia, propofol was superior to methohexital, etomidate and thiopental, but equal to sevoflurane and desflurane. Desflurane and sevoflurane were both superior to thiopental. There was no detectable difference between sevoflurane and isoflurane. With respect to the maintenance of anaesthesia, isoflurane and halothane were the worst. There were no significant differences between propofol, desflurane, sevoflurane and enflurane. Propofol is the induction agent of choice in day case patients. The use of a propofol infusion and avoidance of nitrous oxide may help to reduce postoperative nausea and vomiting. PMID- 12553382 TI - Breathing pattern and workload during automatic tube compensation, pressure support and T-piece trials in weaning patients. AB - BACKGROUND AND OBJECTIVE: Automatic tube compensation has been designed as a new ventilatory mode to compensate for the non-linear resistance of the endotracheal tube. The study investigated the effects of automatic tube compensation compared with breathing through a T-piece or pressure support during a trial of spontaneous breathing used for weaning patients from mechanical ventilation of the lungs. METHODS: Twelve patients were studied who were ready for weaning after prolonged mechanical ventilation (10.2 +/- 8.4 days) due to acute respiratory failure. Patients with chronic obstructive pulmonary disease were excluded. Thirty minutes of automatic tube compensation were compared with 30 min periods of 7 cmH2O pressure support and T-piece breathing. Breathing patterns and workload indices were measured at the end of each study period. RESULTS: During T piece breathing, the peak inspiratory flow rate (0.65 +/- 0.20 L s(-1)) and minute ventilation (8.9 +/- 2.7L min(-1)) were lower than during either pressure support (peak inspiratory flow rate 0.81 +/- 0.25 L s(-1) minute ventilation 10.2 +/- 2.3 L min(-1), respectively) or automatic tube compensation (peak inspiratory flow rate 0.75 +/- 0.26L s(-1); minute ventilation 10.8 +/- 2.7 L min(-1)). The pressure-time product as well as patients' work of breathing were comparable during automatic tube compensation (pressure-time product 214.5 +/- 104.6 cmH2O s(-1) min(-1), patient work of breathing 1.1 +/- 0.4 J L(-1)) and T-piece breathing (pressure-time product 208.3 +/- 121.6 cmH2O s(-1) min(-1), patient work of breathing 1.1 +/- 0.4 J L(-1)), whereas pressure support resulted in a significant decrease in workload indices (pressure-time product 121.2 +/- 64.1 cmH2O s(-1) min(-1), patient work of breathing 0.7 +/- 0.4 J L(-1)). CONCLUSIONS: In weaning from mechanical lung ventilation, patients' work of breathing during spontaneous breathing trials is clearly reduced by the application of pressure support 7 cmH2O, whereas the workload during automatic tube compensation corresponded closely to the values during trials of breathing through a T-piece. PMID- 12553383 TI - Trendelenburg positioning after cardiac surgery: effects on intrathoracic blood volume index and cardiac performance. AB - BACKGROUND AND OBJECTIVE: The efficacy of the Trendelenburg position, a common first step to treat suspected hypovolaemia, remains controversial. We evaluated its haemodynamic effects on cardiac preload and performance in patients after cardiac surgery. METHODS: Twelve patients undergoing mechanical ventilation of the lungs who demonstrated left ventricular 'kissing papillary muscles' by transoesophageal echocardiography, thus suggesting hypovolaemia, were positioned 30 degrees head down for 15 min immediately after cardiac surgery. Cardiac output by thermodilution, central venous pressure, pulmonary artery occlusion pressure, left ventricular end-diastolic area by transoesophageal echocardiography and intrathoracic blood volume by thermo- and dye dilution were determined before, during and after this Trendelenburg manoeuvre. RESULTS: Trendelenburg's manoeuvre was associated with increases in central venous pressure (9 +/- 2 to 12 +/- 3 mmHg) and pulmonary artery occlusion pressure (8 +/- 2 to 11 +/- 3 mmHg). The intrathoracic blood volume index increased slightly (dye dilution from 836 +/- 129 to 872 +/- 112 mL m(-2); thermodilution from 823 +/- 129 to 850 +/- 131 mL m( 2)) as did the left ventricular end-diastolic area index (7.5 +/- 2.1 to 8.1 +/- 1.7 cm2 m(-2)), whereas mean arterial pressure and the cardiac index did not change significantly. After supine repositioning, the cardiac index decreased significantly below baseline (3.0 +/- 0.6 versus 3.5 +/- 0.8 L min(-1) m(-2)) as did mean arterial pressure (76 +/- 12 versus 85 +/- 11 mmHg), central venous pressure (8 +/- 2 mmHg) and pulmonary artery occlusion pressure (6 +/- 4 mmHg). The intrathoracic blood volume index and left ventricular end-diastolic area index did not differ significantly from baseline. CONCLUSIONS: Trendelenburg's manoeuvre caused only a slight increase of preload volume, despite marked increases in cardiac-filling pressures, without significantly improving cardiac performance. PMID- 12553384 TI - Increasing the injection volume by dilution improves the onset of motor blockade, but not sensory blockade of ropivacaine for brachial plexus block. AB - BACKGROUND AND OBJECTIVE: Ropivacaine used for axillary plexus block provides effective motor and sensory blockade. Varying clinical dosage recommendations exist. Increasing the dosage by increasing the concentration showed no improvement in onset. We compared the behaviour of a constant dose of ropivacaine 150 mg diluted in a 30, 40 or 60 mL injection volume for axillary (brachial) plexus block. METHODS: A prospective, randomized, observer-blinded study on patients undergoing elective hand surgery was conducted in a community hospital. Three groups of patients with a constant dose of ropivacaine 150 mg, diluted in 30,40 or 60 mL NaCl 0.9%, for axillary plexus blockade were compared for onset times of motor and sensory block onset by assessing muscle strength, two-point discrimination and constant-touch sensation. RESULTS: Increasing the injection volume of ropivacaine 150 mg to 60 mL led to a faster onset of motor block, but not of sensory block, in axillary plexus block, compared with 30 or 40 mL volumes of injection. CONCLUSIONS: The data show that the onset of motor, but not of sensory block, is accelerated by increasing the injection volume to 60 mL using ropivacaine 150 mg for axillary plexus block. This may be useful for a more rapid determination of whether the brachial plexus block is effective. However, when performing surgery in the area of the block, sensory block onset seems more important. PMID- 12553385 TI - Anaesthetic and haemodynamic effects of continuous spinal versus continuous epidural anaesthesia with prilocaine. AB - BACKGROUND AND OBJECTIVE: To compare, using prilocaine, the effects of continuous spinal anaesthesia (CSA) and continuous epidural anaesthesia (CEA) on haemodynamic stability as well as the quality of anaesthesia and recovery in patients undergoing transurethral resection of the prostate gland. METHODS: Thirty patients (>60 yr) were randomized into two groups. Prilocaine, 2% 40 mg, was given to patients in the CSA group, and prilocaine 1% 150mg was given to patients in the CEA group. Incremental doses were given if the level of sensory block was lower than T10 or if needed during surgery. RESULTS: There was a significant decrease in mean arterial pressure in Group CEA compared with Group CSA (P < 0.01). The decrease in heart rate in Group CSA occurred 10 min after the first local anaesthetic administration and continued through the operation (P < 0.05). The level of sensory anaesthesia was similar in both groups. The times to reach the level of T10 and the upper level of sensory blockade (Tmax) were 18.0 +/- 4.7 and 25.3 +/- 7.0 min in Groups CSA and CEA, respectively, and were significantly longer in Group CEA. The duration of anaesthesia was 76.8 +/- 4min and was shorter in Group CSA (P < 0.01). CONCLUSIONS: Spinal or epidural anaesthesia administered continuously was reliable in elderly patients undergoing transurethral resection of the prostate. Continuous spinal anaesthesia had a more rapid onset of action, produced more effective sensory and motor blockade and had a shorter recovery period. Prilocaine appeared to be a safe local anaesthetic for use with either continuous spinal anaesthesia or continuous epidural anaesthesia. PMID- 12553386 TI - Prediction of difficult tracheal intubation. AB - BACKGROUND AND OBJECTIVE: Preoperative bedside screening tests for difficult tracheal intubation may be neither sensitive nor specific enough for clinical use. The aim was to investigate if a combination of the Mallampati classification of the oropharyngeal view with either the thyromental or sternomental distance measurement improved the predictive value. METHODS: A total of 212 (109 male, 103 female) non-obstetric surgical patients, aged >18 yr, undergoing elective surgical procedures requiring tracheal intubation were assessed preoperatively with respect to the oropharyngeal (modified Mallampati) classification, thyromental and sternomental distances. An experienced anaesthetist, blinded to the preoperative airway assessment, performed laryngoscopy and graded the view according to Cormack and Lehane's classification. RESULTS: Twenty tracheal intubations (9%) were difficult as defined by a Cormack and Lehane Grade 3 or 4, or the requirement for a bougie in patients with Cormack and Lehane Grade 2. Used alone, the Mallampati oropharyngeal view, and thyromental and sternomental distances were associated with poor sensitivity, specificity and positive predictive values. Combining the Mallampati Class III or IV with either a thyromental distance <6.5cm or a sternomental distance <12.5cm decreased the sensitivity (from 40 to 25 and 20%, respectively), but maintained a negative predictive value of 93%. The specificity and positive predictive values increased from 89 and 27% respectively for Mallampati alone to 100%. CONCLUSIONS: The findings suggest that the Mallampati classification, in conjunction with measurement of the thyromental and sternomental distances, may be a useful routine screening test for preoperative prediction of difficult tracheal intubation. PMID- 12553387 TI - Cortical activity assessed by Narcotrend in relation to haemodynamic responses to tracheal intubation at different stages of cortical suppression and reflex control. AB - BACKGROUND AND OBJECTIVE: Many anaesthesiologists still interpret haemodynamic responses as signs of insufficient cortical suppression. The aim was to illustrate how haemodynamics may only poorly reflect the level of cortical suppression and that electroencephalographic monitoring could indicate different relationships between cortical effects and haemodynamics. METHODS: Anaesthesia was induced with thiopental (7 mg kg(-1)), and fentanyl (2 microg kg(-1)) with succinylcholine (1.5 mg kg(-1)) for neuromuscular blockade in the 11 patients of Group 1. In Group 2 (n = 15), thiopental (7 mg kg(-1)) and succinylcholine (1.5 mg kg(-1)) were given. In Group 3, the patients (n = 13) received thiopental (7 mg kg(-1)), fentanyl (2 microg kg(-1)) and cisatracurium (0.1 mg kg(-1)), and they were intubated 3 min later than the patients in Groups 1 and 2. We determined conventional electroencephalographic (EEG) variables and classified 14 EEG stages in real-time ranging from A (= 1), indicating full wakefulness, to F1 (= 14), at profound cortical suppression. RESULTS: All groups had profound cortical suppression 45 s after thiopental administration, which rapidly decreased (EEG stage, 11 (6-13) versus 7 (2-13) at 4 min, P < 0.0001). Decreasing EEG stages were associated with increasing SEF 95, relative alpha and beta power and decreasing relative delta power. During tracheal intubation, profound cortical suppression remained unchanged in Groups 1 and 2. In Group 3, cortical suppression had decreased before laryngoscopy (P < 0.005). In Group 2, 11 patients had heart rate responses to tracheal intubation, whereas only two responded in Group 1 (P = 0.015) and three in Group 3 (P = 0.02). Thirteen patients in Group 2 had arterial pressure responses, and five in Group 1 (P = 0.038). Circulatory responses did not differ between Groups 1 and 3. CONCLUSIONS: Electroencephalographic monitoring was suitable to indicate in real-time that haemodynamics only poorly reflect rapidly changing levels of cortical suppression, and how haemodynamics and cortical activity depend on the applied combination of hypnotic and analgesic drugs during anaesthesia induction with thiopental. PMID- 12553388 TI - Cricoid yoke: the effect of surface area and applied force on discomfort experienced by conscious volunteers. AB - BACKGROUND AND OBJECTIVE: The application of cricoid force is central to techniques that reduce the risk of gastric regurgitation and the subsequent pulmonary aspiration associated with obstetric and emergency anaesthesia. The discomfort associated with cricoid force in awake preoperative patients increases the incidence of coughing, struggling and pain during induction of anaesthesia. This study determined if increasing the surface area of a cricoid yoke reduced the associated discomfort in volunteers. METHODS: Fifty volunteers participated in a randomized single-blinded study. The cricoid yoke was positioned using standard anatomical landmarks and forces of 10, 20, 30 and 40 N were applied in a random order for 20s, using two different yoke attachments with surface areas of 3 and 10 cm2. A rest of 30s was allowed between the application of forces. Discomfort was graded by volunteers on a scale from 0 to 10 (0: no discomfort; 10: worse discomfort imaginable). A score of 10 was allocated if the volunteers could not tolerate the applied force for 20s. RESULTS: Median scores for the small yoke were always higher than those for the large yoke at each force. There were significant differences between the scores for the small and large yokes at 10 and 20 N (P < 0.001) and 30 N (P = 0.0233), but there was no significant difference at 40 N. CONCLUSIONS: The larger yoke was tolerated better by volunteers when clinically relevant cricoid forces were applied. PMID- 12553389 TI - Effects of different doses of oral ketamine for premedication of children. AB - BACKGROUND AND OBJECTIVE: A need exists for a safe and effective oral preanaesthetic medication for use in children undergoing elective surgery. The study sought to define the dose of oral ketamine that would facilitate induction of anaesthesia without causing significant side-effects. METHODS: We studied 80 children undergoing elective surgery under general anaesthesia who received oral ketamine 4, 6 or 8 mg kg(-1) in a prospective, randomized, double-blind placebo controlled study. We compared the reaction to separation from parents, transport to the operating room, the response to intravenous cannula insertion and application of an anaesthetic facemask, the induction of anaesthesia and recovery from anaesthesia. RESULTS: In the group receiving ketamine 8 mg kg(-1), the children were significantly calmer than those of the other groups, and anaesthesia induction was more comfortable. Recovery from anaesthesia was longer in the group receiving ketamine 8 mg kg(-1) compared with the other groups, but no differences between the groups were observed after 2 h in the recovery room. CONCLUSIONS: It is concluded that oral ketamine 8 mg kg(-1) is an effective oral premedication in inpatient children undergoing elective surgery. PMID- 12553390 TI - Remifentanil versus alfentanil in total intravenous anaesthesia for day case surgery. AB - BACKGROUND AND OBJECTIVE: We assessed the intraoperative haemodynamic responses and recovery profiles of total intravenous anaesthesia with remifentanil and alfentanil for outpatient surgery. METHODS: Patients in Group 1 (n = 20) received alfentanil 20 microg kg(-1) followed by 2 microg kg(-1) min(-1) intravenously; patients in Group 2 (n = 20) received remifentanil 1 microg kg(-1) followed by 0.5 microg kg(-1) min(-1) intravenously. Both groups then received propofol 2 mg kg(-1) followed by 9 mg kg(-1) h(-1) intravenously. Five minutes after skin incision, infusion rates were decreased, and at the end of surgery, all infusions were discontinued. Early recovery was assessed by the Aldrete score, whereas intermediate recovery was assessed with the postanaesthetic discharge scoring system (PADS). RESULTS: Perioperative arterial pressure was similar in both groups; heart rate was lower in Group 2 (P < 0.05). The times to spontaneous and adequate respiration, response to verbal commands, extubation and times for Aldrete score > or = 9 were shorter in Group 2 patients (P < 0.05). Pain scores were higher in Group 2 patients (P < 0.05). Overall times for postanaesthetic discharge scores > or = 9 were similar. CONCLUSIONS: Early recovery of patients after day surgery is significantly shorter after total intravenous anaesthesia with remifentanil compared with that with alfentanil but postoperative pain management must be planned ahead. PMID- 12553391 TI - Stability of the LMA-ProSeal and standard laryngeal mask airway in different head and neck positions: a randomized crossover study. AB - BACKGROUND AND OBJECTIVE: The LMA-ProSeal laryngeal mask airway is a new laryngeal mask airway with a modified cuff and drainage tube. We compared oropharyngeal leak pressure, intracuff pressure and anatomical position (assessed fibreoptically) for the Size 5 LMA-ProSeal laryngeal mask airway and the classic laryngeal mask airway in different head-neck positions and using different intracuff inflation volumes. METHODS: Thirty paralysed anaesthetized adult male patients were studied. The LMA-ProSeal laryngeal mask airway and the classic laryngeal mask airway were inserted into each patient in random order. The oropharyngeal leak pressure, intracuff pressure, and anatomical position of the airway tube and drainage tube (LMA-ProSeal laryngeal mask airway only) were documented in four head and neck positions (neutral first, then flexion, extension and rotation in random order), and at 0-40 mL cuff volumes in the neutral position in 10 mL increments. RESULTS: Compared with the neutral position, the oropharyngeal leak pressure for both the LMA-ProSeal laryngeal mask airway and the classic laryngeal mask airway was higher in flexion and rotation (all P < or = 0.02), but lower in extension (all P < or = 0.01). Changes in head neck position did not alter the anatomical position of the airway tube or the drainage tube. The oropharyngeal leak pressure was always higher for the LMA ProSeal laryngeal mask airway (all P < or = 0.005) and anatomical position better for the classic laryngeal mask airway (all P < or = 0.04). CONCLUSIONS: The anatomical position of the LMA-ProSeal and the classic laryngeal mask airway is stable in different head-neck positions, but head-neck flexion and rotation are associated with an increase, and head-neck extension a decrease, in oropharyngeal leak pressure and intracuff pressure. The Size 5 LMA-ProSeal laryngeal mask airway is capable of forming a more effective seal than the Size 5 classic laryngeal mask airway in males. PMID- 12553392 TI - Blood pressure control with glyceryl trinitrate during electroconvulsive therapy in a patient with cerebral aneurysm. PMID- 12553393 TI - Failure of interscalene brachial plexus blockade to produce pre-emptive analgesia after shoulder surgery. PMID- 12553394 TI - Laryngeal mask airway severed by biting. PMID- 12553395 TI - Sickle cell disease in pregnancy. PMID- 12553396 TI - General anaesthesia with remifentanil and cisatracurium for a superobese patient. PMID- 12553397 TI - Antimicrobial activity of garlic, tea tree oil, and chlorhexidine against oral microorganisms. AB - OBJECTIVE: To compare the antimicrobial activity of tea tree oil, garlic, and chlorhexidine solutions against oral microorganisms. METHOD: The five-week study consisted of thirty subjects. The first week was considered baseline. All subjects used a control solution (second week), and were randomly divided into the three groups (third week): G1-0.12% chlorhexidine; G2 - 2.5% garlic (Allium sativum, L.); and G3 - 0.2% tea tree oil (Melaleuca alternifolia). Dishes containing blood agar and Mitis Salivarius Bacitracin agar (MSB) were inoculated with the subjects' saliva (collected twice a week). Total microorganisms and mutans streptococci were counted in blood agar and MSB, respectively. RESULTS: Chlorhexidine and garlic groups showed antimicrobial activity against mutans streptococci, but not against other oral microorganisms. The tea tree oil group showed antimicrobial activity against mutans streptococci and other oral microorganisms. Maintenance of reduced levels of microorganisms was observed only for garlic and tea tree oil during the two consecutive weeks (fourth and fifth). Unpleasant taste (chlorhexidine 40%, tea tree oil 30%, garlic 100%), burning sensation (chlorhexidine 40%, tea tree oil 60%, garlic 100%), bad breath (chlorhexidine 40%, tea tree oil 20%, garlic 90%), and nausea (chlorhexidine 0%, tea tree oil 10%, garlic 30%) were reported. CONCLUSION: Garlic and tea tree oil might be an alternative to chlorhexidine. PMID- 12553398 TI - Prevalence of dental caries in Sri Lankan aboriginal Veddha children. AB - OBJECTIVE: To evaluate the cares status of Sri-Lankan Veddha children in relation to the traditional risk factors. Prevalence of dental caries is still high in certain populations, studies of which may shed additional light on the complex aetiology of caries. METHODS: Using the NIDR criteria for caries diagnosis, the Strip Mutans technique (Dentocult SM), and a questionnaire administered by a native speaker, 39 children between the ages of 2-17 years were examined in a cross-sectional study. RESULTS: Of the 2-9-year-olds, 33% were caries free in the primary dentition and 72% of 5-17-year-olds were caries free in the permanent dentition (mean dft = 2.7; SE = 0.61 and mean DMFT = 0.9; SE = 0.41). There was no correlation between caries and mutans streptococci levels. Excessive consumption of sugar or honey was significantly correlated with the mean dft levels (P = 0.007). The mother was the primary caregiver for all but two children and the children were breast-fed for an average of 2.8 years (SD = 0.82). The duration of breast-feeding was not correlated with the caries experience. CONCLUSION: Lack of professional and home care and unfavourable dietary practices may be related to the high prevalence of caries observed in this population. PMID- 12553399 TI - Restoration of teeth with more-viscous glass ionomer cements following radiation induced caries. AB - OBJECTIVE: To assess the suitability of more-viscous conventional restorative glass ionomer cements (GICs) in a high-caries risk group of patients. METHODS: Fifteen adult patients with radiation-induced caries were treated at a dental hospital by one dentist. Two encapsulated aesthetic GICs were used in each patient to restore 146 carious lesions in the exposed dentine and cementum of 93 teeth. The restorations were assessed directly over two years for their retention, secondary caries, anatomic form, marginal integrity, marginal discolouration, and surface texture. RESULTS: Both GICs were placed in similar sized cavities (P = 0.63). After two years, although 30.0% of Ketac-Molar Aplicap and 12.5% of Fuji IX GP restorations had been lost (P = 0.01), there were no instances of secondary caries. The remaining GICs showed ongoing marginal deterioration, but there were very few instances where this required the repair or replacement of the restorations. No restorations failed from surface erosion. CONCLUSIONS: In these high-caries risk patients the placement of more-viscous GICs appeared to prevent secondary caries, even when the restorations were subsequently lost. PMID- 12553400 TI - Disparities in self reported oral health problems among a young Syrian adult population. AB - OBJECTIVE: To describe the prevalence of dental pain and other oral health problems among a young Syrian adult population and to identify socio-demographic factors associated with these experiences. SUBJECTS: An industrial sample of 400 men and women aged 18-34 years from Damascus, Syria. METHODS: Study participants were interviewed about their experience of oral health problems in the previous year (1998). Socio-demographic information was collected. RESULTS: 93% (369) of the interviews were completed. The prevalence of oral health problems was high, with 96% (353) of respondents claiming that they experienced one or more problems in the previous year. Two thirds of participants (65%, 239) claimed they had dental pain in the previous year. Analysis revealed that dental pain experience was significantly associated with age and gender. Analysis considering all factors revealed that the odds of experiencing dental pain were higher amongst the younger age group (18-24 compared to 25-34 year-olds). CONCLUSION: The prevalence of oral health problems was high among the population studied. Socio demographic variations in experience of dental pain were apparent, with young men of lower education having the greatest odds of dental pain experience in the previous year. PMID- 12553401 TI - Caries experience of 5-6-year-old and 12-13-year-old schoolchildren in central and western Nepal. AB - OBJECTIVE: To describe and analyse the caries experience and caries prevalence in the deciduous dentition of 5-6-year-old schoolchildren and in the permanent dentition of 12-13-year-old schoolchildren in western and central Nepal. DESIGN: Non-randomised cross-sectional surveys conducted by trained and calibrated examiners. SETTING: Surveys were conducted in private and government rural and urban schools at 10 sites along the Terai and the foothills in eight districts of western and central Nepal. SUBJECTS: A total of 2,177, 5-6-year-old and 3,323, 12 13-year-old schoolchildren from urban and rural areas were examined under WHO Pathfinder methodology. OUTCOME MEASURES: Prevalence of caries and dental caries experience (dmft/DMFT). RESULTS: The caries prevalence and mean dmft score of 5-6 year-olds was 67% and 3.3 (urban 64% and 2.9; rural 78% and 4.0). The caries prevalence and mean DMFT score of 12-13-year-olds was 41% and 1.1 (urban 35% and 0.9; rural 54% and 1.5). The d/D-component constituted almost the entire dmft/DMFT index. CONCLUSION: The recorded prevalence of untreated dental caries in schoolchildren requires an appropriate oral health response based primarily on prevention and health promotion. Foremost in this regime would be the promotion and use of accessible and affordable fluoridated toothpaste. PMID- 12553402 TI - Composite resin restorations: a cross-sectional survey of placement and replacement in Jordan. AB - AIMS: To record the reasons for first time placement and replacement of composite resin restorations in Jordan, to determine the use of composite resin restorations in common cavity types and to collect data on the age of the replaced composite resin restorations. SUBJECTS AND SETTING: Dentists in Jordan (n = 241). METHOD: Cross- sectional study using postal survey backed up with personal contact. Data were recorded for all restorations placed or replaced over a period of one month. RESULTS: Information was collected on 2,239 restorations from patients aged 12-65 years. Of all restorations, 61.6% were first time placements while 38.4% were replacement of old restorations. The major reason for the first time placement of restorations was primary caries while that for replacement was secondary caries 36.2%, followed by root canal therapy 22.2%, discolouration 14.4%, lost restorations 13.4%, composite fracture 11.3%, pain or sensitivity 2.4%. CONCLUSIONS: The main reason for first time placement of composite restorations is primary caries; the main reason for replacement is secondary caries. PMID- 12553403 TI - Sources of oral health information and teaching materials for primary schoolteachers in Rungwe District, Tanzania. AB - OBJECTIVE: To determine the various major sources of Oral Health Education (OHE) for primary school teachers in Rungwe District, Mbeya Region, Tanzania. DESIGN: Cross-sectional study using a self-administered close-ended questionnaire. PARTICIPANTS AND SETTING: 232 rural and urban in-service primary school teachers and 195 final-year-teacher trainees at college. MAIN OUTCOME MEASURES: Socio demographic factors, common teachers' OHE sources, common sources of OHE teaching aides, and teachers' previous OHE training opportunity. RESULTS: Textbooks and a dental clinic were found to be the most common sources of OHE for a large proportion of in-service and trainee teachers, while neighbours and a teachers' college were reported by a smaller proportion. The District Dental Officer was the only significant source of oral health education teaching materials in this study. CONCLUSION AND RECOMMENDATION: While primary schoolteachers' sources for oral health education information was mainly textbooks and from a dental clinic, the source of teaching materials was largely a District Dental Officer. Details on the particular sources were not sought and further studies are therefore required. However, the findings highlight a need for standardised and equally distributed oral health material sources both for valid information and for teaching in schools. PMID- 12553404 TI - A study on the adhesion of restorative materials to irreversible hydrocolloid impression materials. AB - AIM: The purpose of this study was to evaluate the adhesion of restorative materials to irreversible hydrocolloid impression materials. MATERIALS AND METHODS: Three irreversible hydrocolloid impression materials were tested with four restorative materials: a base metal alloy, an amalgam, an autopolymerising resin composite and a heat polymerising resin. Samples were prepared with and without a collar. Half of the samples were cleaned with methanol by wiping. The other half were not methanol-wiped. All the samples were submerged into the impression materials and were subsequently placed in an impression- holding device in two different positions. A pullout test using a Hounsfield machine was used after the impression materials were set. RESULTS: Variance analysis was used in comparing the results obtained. Statistically, the four restorative materials showed different values regarding the adherence of the impressions materials to them (composite resin 52.47N, metal 26.36N). Samples with adherence to a collar were found to be more retentive than those without a collar. Samples with impression materials with a full collar were more retentive than those without a collar, and the samples cleaned with methanol were again less retentive than those which were not methanol wiped. CONCLUSION: The results of this study show that adhesion may be seen between restorative materials and irreversible hydrocolloid impression materials, and this adhesion may affect the accuracy of the impression. PMID- 12553405 TI - A comparison of the psychosocial environment of two types of residences for persons with severe mental illness: small congregate community residences and psychiatric inpatient settings. AB - BACKGROUND: Legislation was passed in Sweden to stimulate the development of different housing solutions for persons suffering from severe mental illness. Among these solutions are small congregate residences built in the community to provide suitable housing and support for those not able or willing to live independently. AIMS: The general aim of the present study was to compare the psychosocial environment of two types of residences for the persons with severe mental illness - congregate community residences and psychiatric inpatient settings. A specific aim was to see if the former provided higher levels of autonomy than the latter. METHODS: In this cross-sectional study the real version of the Community Oriented Program Environmental Scale (COPES) was administered to residents, patients and staff members. RESULTS: The study showed that the psychosocial environment differs between the two types of settings. Residents and staff in small congregate residences rated higher levels of Autonomy and lower levels of Practical Orientation, Anger and Aggression and Order and Organisation than patients and staff in inpatient settings. CONCLUSIONS: The psychosocial environment profiles for psychiatric settings in different phases of the care process may vary in terms of the aforementioned subscales. PMID- 12553406 TI - A mutual-aid group for psychiatric rehabilitation of mental ex-patients in Hong Kong. AB - BACKGROUND: Mutual-aid group and self-directed group are recognized as important interventions for anti-oppression of disadvantaged groups in our society. METHOD: In this paper, the writer describes a mutual-aid group implemented in a psychiatric half-way house in Hong Kong. Members in this group were free to set their own agenda for discussion and to invite guest speakers. In terms of various stages of this group, the writer discusses how different group activities facilitated the development of mutual-aid and self-directed natures of mental ex patients. RESULTS: The findings show that this group can have an impact on the wider community and aid in improving the self-image of mental ex-patients and the perception of them as contributing positively to that community, hence lessening negative stereotypes. PMID- 12553407 TI - A comparison of Russian and British attitudes towards mental health problems in the community. AB - AIMS & RESULTS: This study examines the differences in attitudes towards mental health problems of Russian and British communities. A specially developed questionnaire containing four vignettes conforming to specific DSM-IV diagnoses of common mental disorders (obsessive compulsive disorder, schizophrenia, depression and dementia) was given to 134 subjects. Additional questions aimed to identify different attitudes towards the mentally ill, public awareness of the possible causes of these disorders and the availability of potential help. The comparisons were made in terms of national differences and demographic characteristics such as sex, age, education, marital status, children and familiarity with mental illness. CONCLUSIONS: The study revealed that the British sample was significantly more tolerant than the Russian. The Russian participants were far less likely to identify dementia as a mental disorder. However, the British respondents chose medically related help significantly more than the Russians. Significant associations were observed between factors such as education and familiarity with mental illness and tolerance within the British group. Various arguments are presented for the observed differences in terms of historic, political and cultural perspectives. PMID- 12553408 TI - Social disablement among residents of hostels for the homeless in Rio de Janeiro, Brazil. AB - BACKGROUND: Knowledge of the extent of social disablement among homeless people is very useful for policy-makers and mental health professionals who are involved in the delivery of community-based care. As there are few studies of this issue in Brazil or other developing countries, a cross-sectional study was carried out among homeless people from the Rio de Janeiro (RJ) metropolitan area. METHOD: Data were collected on a random sample of 330 residents in the five public hostels using the 21-items Social Behaviour Scale (SBS) and the Composite International Development Interview (CIDI). RESULTS: The mean age of the 330 people in the sample was 44.8, 75.8% of them were men, 78.9% were single, 79.2% unemployed. Ten percent of the 330 residents had more than five social behaviour problems, but 39.4% of them had no problem. Some 26.7% were scored as under active, 23.0% with poor self-care, 13.3% with slowness, 12.7% with restlessness and 11.8% inappropriate social mixing. Those with schizophrenia were reported to have many more disabilities in almost all of the SBS areas. On the other hand, depression and hostility were more frequent in those with diagnoses of depression and substance abuse disorders. Women were more likely to be reported as having disablement than men, explained by their greater prevalence rate of schizophrenia. CONCLUSIONS: In this population, social disablement was largely explained by those with schizophrenia. Active treatment strategies are needed for this group. Other residents had few behaviour problems, their most pressing problems come from their poor social circumstances. PMID- 12553409 TI - Suicide amongst Irish migrants in Britain: a review of the identity and integration hypothesis. AB - BACKGROUND: Studies have consistently reported higher rates of suicide amongst Irish migrants in Britain than in the population as a whole. Leavey offers a hypothetical model to explain such rates that incorporates lack of social cohesion and integration meshed with the inability to establish an authentic identity and other contributory factors. MATERIAL: Systematic review methodologies are used to examine the central tenets of this explanatory framework. Some of the macro-level ecological associations in the model are critically evaluated in the context of findings from the 1991 Census and government social and household panel surveys. DISCUSSION: The evidence base suggests that statements on social isolation and reluctance to use health care services are questionable and Irish migration is shown to be much more heterogeneous than the model suggests. Only small positive, and as yet unreplicated, associations have been established between identity and health behaviour in a non-representative sample and evidence is lacking of Irish stoicism and anti-Irish racism as putative risk factors. Epidemiological studies show that adjusting suicide rates for social class explains virtually none of the excess in Irish migrants, although higher risks for unmarried persons are reported. Explanations in the literature for higher rates of migrant suicide are discussed. CONCLUSIONS: Studies based on individual-level analysis and record linkage are urgently needed to explain the high rates. PMID- 12553410 TI - Perception of and attitude towards mental illness in Oman. AB - BACKGROUND: As conceptions of mental illness are often dictated by prevailing socio-cultural factors and the philosophy of the time, there is little research to substantiate how mental illness is perceived in the Arab world in the light of both traditional and more recent modernization and acculturation processes. AIMS: To examine whether social factors exert an influence on a person's attitude towards people with mental illness (PWMI) in the rapidly changing country of Oman. METHODS: This study compares the response elicited from medical students, relatives of psychiatric patients and the general Omani public on the causes of mental illness, attitudes toward PWMI and the care and management of people with mental illness. RESULTS: This study found no relationship between attitudes towards PWMI, and demographic variables such as age, educational level, marital status, sex and personal exposure to people with mental illness. Both medical students and the public rejected a genetic factor as the cause of mental illness; instead they favoured the role of spirits as the aetiological factor for mental illness. There were favourable responses on statements regarding value of life, family life, decision-making ability, and the management and care of mental illness. However, both medical students and the public thought that PWMI tend to have peculiar and stereotypical appearances and the majority preferred that facilities for psychiatric care should be located away from the community. Although the relatives of psychiatric patients were concerned about the welfare of mental patients, their responses varied and were often contingent upon their expectations. CONCLUSIONS: The data suggest that neither socio-demographic factors nor previous exposure to PWMI was related to attitudes towards PWMI. Although the attitudes of Omanis toward PWMI appear to fluctuate in complex ways, traditional beliefs on mental illness have yet to be eroded by exposure to a biomedical model of mental illness. This study largely supports the view that the extent of stigma varies according to the cultural and sociological backgrounds of each society. PMID- 12553411 TI - Personality differences in childhood and adolescence: measurement, development, and consequences. AB - Child psychologists and psychiatrists are interested in assessing children's personalities. This interest is fueled by the practical desire to identify differences between children that have predictive utility, and by recognition that future advances in developmental theory, especially in relation to gene environment interplay, can only be as good as the measures on which they rely. The aim of this article is to facilitate these practical and theoretical advances. First, we delineate a taxonomy of measurable individual differences in temperament and personality in childhood, and point the reader to proven and/or promising measuring instruments. Second, we describe the processes through which early temperament differences may become elaborated into adult personality structure and lifelong adaptation, and identify gaps in the empirical research that need to be filled. Third, we explore the various connections between temperament/ personality traits and psychopathology, and direct attention to promising questions and strategies. PMID- 12553412 TI - Experience effects on brain development: possible contributions to psychopathology. AB - Researchers and clinicians are increasingly recognizing that psychological and psychiatric disorders are often developmentally progressive, and that diagnosis often represents a point along that progression that is defined largely by our abilities to detect symptoms. As a result, strategies that guide our searches for the root causes and etiologies of these disorders are beginning to change. This review describes interactions between genetics and experience that influence the development of psychopathologies. Following a discussion of normal brain development that highlights how specific cellular processes may be targeted by genetic or environmental factors, we focus on four disorders whose origins range from genetic (fragile X syndrome) to environmental (fetal alcohol syndrome) or a mixture of both factors (depression and schizophrenia). C.H. Waddington's canalization model (slightly modified) is used as a tool to conceptualize the interactive influences of genetics and experience in the development of these psychopathologies. Although this model was originally proposed to describe the 'canalizing' role of genetics in promoting normative development, it serves here to help visualize, for example, the effects of adverse (stressful) experience in the kindling model of depression, and the multiple etiologies that may underlie the development of schizophrenia. Waddington's model is also useful in understanding the canalizing influence of experience-based therapeutic approaches, which also likely bring about 'organic' changes in the brain. Finally, in light of increased evidence for the role of experience in the development and treatment of psychopathologies, we suggest that future strategies for identifying the underlying causes of these disorders be based less on the mechanisms of action of effective pharmacological treatments, and more on increased knowledge of the brain's cellular mechanisms of plastic change. PMID- 12553413 TI - Non-human primate models of childhood psychopathology: the promise and the limitations. AB - Although non-human primate models have been used previously to investigate the neurobiology of several sensory and cognitive developmental pathologies, they have been employed only sparingly to study the etiology of childhood psychopathologies for which deficits in social behavior and emotion regulation are major symptoms. Previous investigations of both adult human and non-human primates have indicated that primate social behavior and emotion are regulated by a complex neural network, in which the amygdala and orbital frontal cortex play major roles. Therefore, this review will provide information generated from the study of macaque monkeys regarding the timing of normal social and emotional behavior development, the normal pattern of anatomical and functional maturation of the amygdala and orbital frontal cortex, as well as information regarding the neural and behavioral effects of early perturbations of these two neural structures. We will also highlight 'critical periods' of macaque development, during which major refinements in the behavioral repertoire appear to coincide with significant neural maturation of the amygdala and/or orbital frontal cortex. The identification of these 'critical periods' may allow one to better predict the specific behavioral impairments likely to appear after neonatal damage to one or both of these neural areas at different time points during development. This experimental approach may provide a new and important way to inform and stimulate research on childhood psychopathologies, such as autism, schizophrenia and Williams syndrome, in which the development of normal social skills and emotional regulation is severely perturbed. Finally, the promise and limitations inherent to the use of non-human primate models of childhood psychopathology will be discussed. PMID- 12553414 TI - Developmental course of ADHD symptomatology during the transition from childhood to adolescence: a review with recommendations. AB - Although historically conceptualized as a disorder that was limited to males during middle childhood, ADHD is currently conceptualized as a chronic disorder that persists into adolescence and adulthood for both sexes. Nonetheless, the veracity of adult ADHD continues to be the source of debate. In order to frame this debate, research leading to the conceptualization of ADHD as a chronic disorder is reviewed. A distinction is made between the developmental outcomes versus the developmental course of ADHD. It is concluded that although childhood ADHD is associated with negative developmental outcomes in adolescence and adulthood, questions about the developmental course of ADHD remain. Although it appears that ADHD diminishes with advancing age, a number of methodological limitations prohibit firm conclusions. Recommendations for future studies are made with an emphasis on 1) overcoming extant methodological limitations in the literature and 2) the need for theoretically derived hypotheses regarding continuity and change in ADHD symptomatology over time. PMID- 12553415 TI - Stress and psychopathology in children and adolescents: is there evidence of specificity? AB - Research on the relations between specific stressors and specific psychological outcomes among children and adolescents is reviewed. Specificity, the notion that particular risk factors are uniquely related to particular outcomes is discussed from a theoretical perspective, and models of specificity are described. Several domains of stressors are examined from a specificity framework (e.g., exposure to violence, abuse, and divorce/marital conflict) in relation to broad-band outcomes of internalizing and externalizing symptoms. Studies that tested for specificity conducted within the past 15 years are examined, and definitional problems are highlighted. Little evidence for specificity was found. Methodological problems in the literature and the lack of theory-driven specificity research are discussed, and directions for future research are identified. PMID- 12553416 TI - Parenting and childhood anxiety: theory, empirical findings, and future directions. AB - Theories of anxiety development suggest that parental acceptance, control, and modeling of anxious behaviors are associated with children's manifestations of anxiety. This paper reviews research published in the past decade on the relation between parenting and childhood anxiety. Observed parental control during parent child interactions was consistently linked with shyness and child anxiety disorders across studies. Mixed support for the role of parental acceptance and modeling of anxious behaviors was found in observational studies. However, there was little evidence supporting the contention that self-reported parenting style was related to children's trait anxiety. Because of limitations associated with past research, inferences about the direction of effects linking parenting and child anxiety cannot be made. A conceptual framework based on recent models of anxiety development (e.g., Vasey & Dadds, 2001) is presented to aid in the interpretation of extant research findings and to provide suggestions for future research and theory development. Improved methodological designs are proposed, including the use of repeated-measure and experimental designs for examining the direction of effects. PMID- 12553417 TI - Historical aspects and perspectives in transthyretin research. PMID- 12553418 TI - Transthyretin from discovery to now. AB - As introduction to the First International Congress on Transthyretin in Health and Disease, this lecture traces the origin of the subjectfrom the discovery in the 1950s that a serum protein migrating ahead of albumin in an electrical field binds the thyroid hormone, thyroxine. Early work defined the molecular and biological properties of thyroxine-binding prealbumin (TBPA). Its tetrameric structure, first recognized from a polymorphism in monkeys, was later elaborated by crystallographic studies, and the very different affinity of its two identical thyroxine-binding sites was explained by an allosteric effect upon occupation of the first site. The far higher concentration of TBPA in cerebrospinal fluid compared to blood was explained by the discovery, 30 years later, that TBPA is synthesized by cells of the choroid plexus, and its rapid turnover in the body made TBPA a convenient marker of malnutrition and chronic disease. Late in the 1960s it was learned that TBPA also carries vitamin A in the circulation by interacting with retinol-binding protein (RBP). TBPA then was renamed transthyretin (TTR), in recognition of its dual transport function, and it was shown that retinol-RBP-TTR interactions are mutually enhancing. Investigation of the molecular genetics of TTR began in 1980 and a large number of inherited variants were discovered in the ensuing years. Some affect thyroxine and/or RBP binding but the majority are associated with familial amyloidotic polyneuropathy. Seizing on this discovery, structural biologists are now investigating why mutated TTR changes from a compact, soluble molecule into a fibrillar, insoluble polymer, and how this pathological transformation might be prevented. PMID- 12553419 TI - The evolution of transthyretin synthesis in vertebrate liver, in primitive eukaryotes and in bacteria. AB - Thyroid hormones are evolutionarily old signal molecules, which can partition between compartments by partitioning into lipid membranes. The role of thyroid hormone distributor proteins is to ensure that sufficient thyroid hormone remains in the bloodstream. Of particular interest is the role of transthyretin, synthesised by the liver and secreted into the blood. In this review, three hypotheses are presented, suggesting the selection pressures leading to the onset of transthyretin synthesis in the liver during evolution. A thyroid hormone distribution network would be a selection advantage over a single protein performing this function. Similarly to the situation in eutherians, hepatic transthyretin synthesis in marsupials is under negative acute phase regulation. The overall three-dimensional structure of transthyretin did not change appreciably during vertebrate evolution. The region of the primary sequence which evolved most was the N-terminal region of the subunit. The N-termini of transthyretin changed from longer and more hydrophobic in reptiles/birds, to shorter and more hydrophilic in eutherians. These changes are correlated with a change in preference from binding of triiodothyronine, to binding thyroxine. As the rest of the molecule had not changed significantly during vertebrate evolution, the gene coding for transthyretin must have evolved prior to the divergence of the vertebrates from the non-vertebrates. Five open reading frames in the genomes of C. elegans (2), S. dublin, S. pombe and E. coli were identified. The protein products are predicted to form tetramers similar to transthyretins. Two possible functions of these proteins are suggested. PMID- 12553420 TI - The evolution of transthyretin synthesis in the choroid plexus. AB - Choroid plexus has the highest concentration of transthyretin (TTR) mRNA in the body, 4.4 microg TTR mRNA/g wet weight tissue, compared with 0.39 microg in the liver. The proportion of TTR to total protein synthesis in choroid plexus is 12%. All newly synthesized TTR is secreted towards the ventricles. Net transfer of T4 occurs only towards the ventricle and depends on ongoing protein synthesis. Thyroxine-binding globulin (TBG), TTR and albumin form a "buffering" system for plasma [T4] because of their overlapping affinities and on/off rates for L thyroxine (T4)-binding. The individual components of this network determining T4 distribution are functionally highly redundant. Absence of TBG (humans), or TTR (mice), or albumin (humans, rats) is not associated with hypothyroidism. Natural selection is based on small, inheritable alterations improving function. The study of these alterations can identify function. TTR genes were cloned and sequenced for a large number of vertebrate species. Systematic, stepwise changes during evolution occurred only in the N-terminal region, which became shorter and more hydrophilic. Simultaneously, a change in function occurred: TTR affinities for T4 are higher in mammals than in reptiles and birds. L-triiodothyronine (T3) affinities show the opposite trend. This favors site-specific regulation of thyroid hormones by tissue-specific deiodinases in the brain. PMID- 12553421 TI - Hepatic synthesis, maturation and complex formation between retinol-binding protein and transthyretin. AB - The retinol/retinol-binding protein/transthyretin complex, that carries and delivers hydrophobic retinol molecules to target cells, is assembled in the hepatocyte endoplasmic reticulum. In this paper, we review data related to events that lead to the formation of this complex, including transthyretin oligomerization and retinol-binding protein secretion. Our studies on transthyretin oligomerization have demonstrated that cleavage of signal peptide and the environment of endoplasmic reticulum influence transthyretin oligomerization. In vitro, mutated transthyretin without signal sequence fails to form dimers, while wild-type transthyretin is translocated into the microsomes where it forms dimers and small amounts of tetramers. In vivo, tetramers were detected in HepG2 cells but not in transfected Cos cells, suggesting that tissue specific factors affect tetramer stability. In vitamin A deficiency, retinol binding protein secretion is blocked and the protein accumulates in the endoplasmic reticulum, from where it is promptly released after retinol repletion. We use MMH cells to identify factors involved in complex formation, retention and secretion, the crucial steps to understand the molecular mechanisms underlying vitamin A homeostasis. In parallel, studies on vitamin A transport in fish are in progress; retinol-binding protein and transthyretin have already been characterized in different fish species. PMID- 12553422 TI - Synthesis and structural analysis of the N-terminal domain of the thyroid hormone binding protein transthyretin. AB - Transthyretin (TTR) is a 55 kDa protein responsible for the transport of thyroid hormones and retinol in human serum. Misfolded forms of the protein are implicated in the amyloid diseases familial amyloidotic polyneuropathy and senile systemic amyloidosis. Its folding properties and stabilization by ligands are of current interest due to their importance in understanding and combating these diseases. To assist in such studies we developed a method for the solid phase synthesis of the monomeric unit of a TTR analogue and its folding to form a functional 55 kDa tetramer. The monomeric unit of the protein was chemically synthesized in three parts, comprising amino acid residues 1-51, 54-99 and 102 127, and ligated using chemoselective thioether ligation chemistry. The synthetic protein was folded and assembled to a tetrameric structure in the presence of the TTR's native ligand, thyroxine, as shown by gel filtration chromatography, native gel electrophoresis, TTR antibody recognition and thyroid hormone binding. In the current study the solution structure of the first of these fragment peptides, TTR(1-51) is examined to determine its intrinsic propensity to form beta-sheet structure, potentially involved in amyloid fibril formation by TTR. Despite the presence of extensive beta-structure in the native form of the protein, the N terminal fragment adopts an essentially random coil conformation in solution. PMID- 12553423 TI - Three-dimensional structure of the transthyretin-retinol-binding protein complex. AB - Transthyretin (TTR), formerly called prealbumin, one of the transporters of the hormone thyroxine and retinol-binding protein (RBP), the specific carrier of vitamin A, forms, under physiological conditions, a macromolecular complex that prevents glomerular filtration of the low-molecular-weight RBP in the kidneys. This paper describes briefly the three-dimensional structure of the two proteins as determined by X-ray diffraction analysis of single crystals and that of the complex that the two molecules form in plasma and discusses the non crystallographic evidence that supports the model of the macromolecular complex. PMID- 12553424 TI - Mechanisms of molecular recognition: crystal structure analysis of human and rat transthyretin inhibitor complexes. AB - Structure-activity data show that many pharmacological agents are strong competitive inhibitors for thyroxine (T4) binding to transthyretin (TTR) and that this competition can interfere with their normal pharmacological actions. TTR is a tetrameric serum protein responsible for the transport of 20% of the circulating T4 in man, while in lower vertebrates such as rats it is the only carrier. The sequence of rat TTR is 85% homologous to the human protein. Crystallographic analyses of ligand co-crystal complexes of human and rat TTR have been studied to understand the molecular basis for binding selectivity of competitor binding to TTR. Analysis of TTR crystal complexes with several classes of competitors (hormone metabolites, flavonoids, fluorescent probes, analgesics and cardiac agents) revealed multiple modes of binding with both forward and reverse ligand binding orientations. These ligands also have different binding positions along the length of the channel with the smallest ligands located deeper within the hormone domain. Data for the human TTR complex with the bromoflavone EMD21388 incubated at different times revealed variable binding positions and occupancies dependent upon incubation time. Comparison of the structures of T4 thyroacetic acid in complex with both human and rat TTR revealed forward and reverse binding, but also showed different modes of binding in the rat compared to the human complex. These data highlight the importance of hydrogen bonding with Lys-15 and Ser-117 and provide insight into ligand binding affinity and negative cooperativity. PMID- 12553425 TI - Transthyretin in fish: state of the art. AB - Relatively little is known about thyroid hormone-binding proteins in fish and, until recently, the thyroid hormones (THs), thyroxine (T4) and triiodothyronine (T3), had only been found in fish plasma bound to albumin and lipoproteins. Recently, transthyretin (TTR) was cloned in a teleost fish, the sea bream (sb); it is composed of 130 amino acids and shares 47-54% sequence similarity with other vertebrate TTR and binds preferentially T3. Homology modelling of sbTTR based upon the crystallographic structure of TTR in human, rat and chicken reveals similar monomer-monomer and dimer-dimer interfaces and a conserved tetrameric structure. In sbTTR, a single amino acid substitution in the thyroid hormone binding site (Ser 117 in human by Thr in sea bream) may explain the higher affinity of this tetramer for T3 rather than T4. The principal site of production of TTR in the sea bream is the liver but transcripts are also present in the intestine, brain, skin, heart, skeletal muscle, kidney, testis, gills and pituitary (in descending order of abundance). The function of TTR in fish remains to be studied but we have recently carried out studies which suggest it may be involved in TH balance during food shortage. PMID- 12553426 TI - The effects of endocrine-disrupting chemicals on thyroid hormone binding to Xenopus laevis transthyretin and thyroid hormone receptor. AB - We investigated the effects of medical, industrial and agricultural chemicals on 3,3',5-L-[125I]triiodothyronine ([125I]T3) binding to purified recombinant Xenopus laevis (X. laevis) transthyretin (xTTR), a plasma thyroid hormone-binding protein, and to the ligand-binding domain of thyroid hormone receptor-beta (xTR LBD). xTTR derived from X. laevis serum had about 80 times higher affinity for T3 than for L-thyroxine. The xTTR's relative affinities for diethylstilbestrol, pentachlorophenol and ioxynil were 10(-1)- to 10(-2)-fold less than that for T3. However, all chemicals investigated had either a weak or no influence on [125I]T3 binding to xTR LBD. The concentration of diethylstilbestrol, the most potent chemical, required for 50% inhibition of [125I]T3 binding to xTR LBD was 10(4) times greater than that of unlabeled T3. These results indicate the existence of several chemicals that interact with xTTR but not with xTR LBD. PMID- 12553427 TI - Clinical picture and outcome of transthyretin-related familial amyloid polyneuropathy (FAP) in Japanese patients. AB - Familial amyloid polyneuropathy (FAP) was once considered to be a disease peculiar to endemic areas, but it is now recognized that FAP kindreds exist in worldwide places. The amyloid precursor of FAP is a variant form of transthyretin (TTR) with one amino acid substitution, which is ascribed to a mutation of TTR gene. Corresponding to the variety of the clinical phenotypes of FAP, more than 80 mutations have been identified as causative gene abnormality in this disease. Since the vast majority of TTR in serum is produced in the liver, liver transplantation has become widely accepted as a valuable treatment for FAP. PMID- 12553428 TI - Laboratory assessment of transthyretin amyloidosis. AB - Mutations in transthyretin (TTR) are the most common cause of autosomal dominant systemic amyloidosis. To date, more than 80 TTR mutations have been associated with amyloidosis in humans. A high prevalence of some mutations like Val122Ile which is identified in 3% of African Americans indicates the necessity of thorough investigation of patients suspected of having, or to be at risk of developing, TTR amyloidosis. Laboratory tests available for evaluation of TTR amyloidosis include both DNA and protein assays. In the case of a known mutation DNA analysis is realized by restriction fragment length polymorphism (RFLP), polymerase chain reaction-induced mutation restriction analysis (PCR-IMRA), single strand confirmation polymorpism (SSCP) or nucleotide sequencing. SSCP, PCR non-isotopic RNAse cleavage assay (NIRCA) or nucleotide sequencing are used to identify an unknown mutation. At the protein level, two techniques are used, isoelectric focusing and mass spectrometry, in both cases (known or unknown mutation). The identification of a previously unknown mutation requires a combination of clinical, pathological and molecular studies. PMID- 12553429 TI - Amyloid: morphology and toxicity. AB - We have expressed transthyretin (TTR) mutants which have significantly destabilised tetramers that aggregate into amyloid fibrils via a series of intermediates. We used atomic force microscopy to follow the morphology of aggregates during fibril formation. Initially, amorphous aggregates are formed that subsequently mature into fibrillar structures. This observation is interpreted as an optimisation of beta-strand registers. The rate of aggregation and maturation is highly temperature-dependent suggesting that entropic forces significantly contribute to stability. In addition, we identified a correlation between the presence of early formed aggregates of TTR and cytotoxicity. The toxic response was mediated via an apoptotic mechanism. The fact that early formed amorphous aggregates, but not more mature fibrils, exert a toxic response suggests that the rate of fibril formation may be a critical parameter. We propose that a slow rate of aggregation facilitates an increased concentration of a toxic intermediate. PMID- 12553430 TI - Historical overview of analytical methods for the measurement of transthyretin. AB - The history of prealbumin dates back to the early forties and may be divided into three parts, based on a chronological and functional approach. The first part- the discovery and the identification of prealbumin--was essentially based on classical protein chemistry methods. The second--the demonstration of prealbumin as a thyroid hormone-binding protein (thyroxine-binding prealbumin)--has greatly benefited from isotopic techniques. The third one--establishing prealbumin as a nutritional marker--was a result of field studies on nutrition. The discovery of the role of prealbumin in retinol binding led to a change in its name, prealbumin becoming transthyretin. Finally, structural studies and mutation analysis of transthyretin in patients with amyloid neuropathy have opened a new area of research. PMID- 12553431 TI - Nutritional regulation of visceral markers in rat liver and cultured hepatoma cells. AB - Protein malnutrition in humans and other animals is consistently associated with a decreased concentration of circulating serum albumin, transthyretin (TTR), and insulin-like growth factor-I (IGF-I). The molecular mechanisms for regulation of the three polypeptides by dietary protein remain to be completely elucidated. The abundance of albumin, TTR and IGF-I mRNA is decreased in liver of juvenile rats consuming insufficient amounts of protein. Moreover, protein restriction specifically decreases the abundance of albumin and TTR nuclear transcripts, indicating that the reduction in mRNA levels for these two genes is caused at least partly by a decrease in gene transcription. Expression of several other genes transcribed at a high level in the liver is also decreased under conditions of dietary protein restriction, suggesting that the level/functional activity of liver-enriched transcription factor(s) might be decreased under these conditions. Limitation of cultured hepatoma cells for a single amino acid also selectively decreases the mRNA levels of several genes with liver-enriched expression, including albumin and TTR. The decrease in albumin mRNA is caused partly by decreased albumin gene transcription and partly by destabilization of albumin mRNA. PMID- 12553432 TI - Significance of transthyretin in protein metabolism. AB - Total body nitrogen (TBN) is mainly sequestered within the metabolically active lean body mass, in close relationship with total body potassium (TBK). TBN and TBK of growing children manifest superimposed accretion rates, display a sexual difference at the onset of adolescence and during adulthood, thereafter decreasing in elderly subjects. Plasma transthyretin (TTR) follows a comparable profile from birth to death in healthy individuals. Uncomplicated protein-energy malnutrition primarily affects the activity of nitrogen metabolic pool, reducing protein syntheses to levels compatible with survival. This adaptive response is well identified by declining TTR concentrations. In various stressful conditions, in vivo responses are characterized by upregulation in injured regions and with muscle proteolysis exceeding protein synthesis, resulting in a net body negative nitrogen balance. Again, this evolutionary pattern mirrors that of plasma TTR. Attenuation of stress and/or introduction of nutritional rehabilitation allows restoration to normal of both TBN and TTR values that follow parallel slopes. Despite distinct etiopathogenic mechanisms, TTR concentrations appear to reflect the loss or gain of TBN in body pools and they predict later outcome in malnutrition and in conditions of acute and/or chronic inflammation. PMID- 12553433 TI - Transthyretin as a thyroid hormone carrier: function revisited. AB - Thyroid hormones are essential for normal mammalian development and for normal metabolism. Thyroxine (T4) is the principal product synthesized by the thyroid follicles, and triiodothyronine (T3), the biologically active hormone, derives mainly from tissue T4 deiodination. More than 99% of the circulating hormone is bound to plasma proteins, mainly to thyroxine-binding globulin, transthyretin and albumin in man, and to transthyretin and albumin in rodents. The role of plasma proteins in the transport of hormones to target tissues has, for a long time, been controversial. The liver and the choroid plexus are the major sites of transthyretin synthesis, tissues from which transthyretin is secreted into the blood and the cerebrospinal fluid, respectively. Transthyretin has been proposed to mediate thyroid hormone transfer into the tissues, particularly into the brain across the choroid-plexus-cerebrospinal fluid barrier. Studies in a transthyretin null mice strain have shown conclusively that transthyretin is not indespensable for thyroid hormones' entry into the brain and other tissues, nor for the maintenance of an euthyroid status. An euthyroid status is also observed in man totally deprived of thyroxine-binding globulin and in rats without albumin. Taken together, these results exclude dependence of thyroid hormone homeostasis on any major plasma carrier per se. This evidence agrees with the free hormone hypothesis which states that the biologically significant fraction, that is taken up by the tissues, is the free circulating hormone. PMID- 12553434 TI - The molar ratio of retinol-binding protein to transthyretin in the assessment of vitamin A status in adults. Proposal of a cut-off point. AB - The molar ratio of retinol-binding protein to transthyretin (RBP:TTR) has been proposed as an indirect method to assess vitamin A status in children with inflammation. Neither reference values nor appropriate cut-off point are available for adults. RBP, TTR and retinol were determined in plasma from 100 healthy adults and 31 low-risk surgical patients with no inflammatory response. RBP:TTR percentile distribution from 99 healthy adults with plasma retinol > or = 0.7 micromol/l was: 2.5th = 0.24; 5th = 0.31; 10th = 0.32; 25th = 0.41; 50th = 0.47; 75th = 0.54; 90th = 0.67; 95th = 0.78 and 97.5th = 0.81. In order to define a cut-off point, receiver operating characteristic (ROC) curve was constructed, using plasma retinol as gold standard. ROC curve was based on data from the 131 studied subjects, 11 of whom (8.4%) were classified as deficient on the basis of plasma retinol < 0.7 pmol/l. According to ROC curve criteria, RBP:TTR ratio was considered a good test, the area under the curve being 0.822, p < 0.001. A cut off-point of < or = 0.37 is proposed to detect vitamin A deficiency in adults, since it allows reaching high sensitivity (81.8%), specificity (79.2%) and predictive value (79.4%). The proposed cut-off point falls between 13th and 14th percentiles. PMID- 12553435 TI - Observations with regard to the National Kidney Foundation K/DOQI clinical practice guidelines concerning serum transthyretin in chronic renal failure. AB - The National Kidney Foundation K/DOQI Guidelines state that, "Serum prealbumin is a valid and clinically useful measure of protein-energy nutritional status in maintenance dialysis (MD) patients." Prealbumin, also known as serum transthyretin (TTR), was not recommended as a nutritional parameter of the same usefulness as the serum albumin. This decision was made, in part, because published research at that time suggested that serum TTR was not a more sensitive index of nutritional status than serum albumin and there was much more clinical and research experience with serum albumin as a nutritional and inflammatory marker. Evidence, including more recently published research data, which is reviewed in this paper has led to the following conclusions by the current authors: 1) In MD patients either protein-energy malnutrition or inflammation can lead to a reduction in serum TTR concentrations. 2) Hence, in MD patients, serum TTR concentrations can be used as a measure of both nutritional and inflammatory status. 3) Serum TTR concentrations are typically increased in MD patients. 4) In maintenance hemodialysis (MHD) patients, serum TTR is a risk factor for mortality that is somewhat independent of serum albumin. 5) Current epidemiological evidence suggests that a serum TTR value of 25 or 30 mg/dl or greater is associated with increased survival and, hence, is desirable in MHD patients. 6) MHD patients with serum TTR levels less than 25-30 mg/dl should be evaluated for protein-energy malnutrition and inflammation. PMID- 12553436 TI - Metabolism and clinical interest of serum transthyretin (prealbumin) in dialysis patients. AB - Chronic renal failure is responsible for an increase in serum concentrations of transthyretin. Elevated serum transthyretin during renal insufficiency is secondary to the lack of retinol-binding protein degradation in renal tubules and to the subsequent increase in the fraction of transthyretin bound to retinol binding protein. In both hemodialysis and peritoneal dialysis patients, serum transthyretin was demonstrated to be a reliable marker of nutritional status, exhibiting significant relationships with energy and protein intakes as well as with fat stores and lean body mass. Serum transthyretin levels less than 300 mg/l were shown to be associated with an increased risk of morbidity and mortality in dialysis patients. The predictive value of transthyretin was shown to be independent of serum albumin. Regular measurements of both serum albumin and transthyretin make it possible to detect patients whose prognosis is compromised by malnutrition and in whom an active nutritional therapy must be undertaken. Simultaneous measurements of inflammatory markers such as serum C-reactive protein are required to evaluate the role of inflammation in serum albumin and transthyretin variations. These low-cost protein parameters should be incorporated in the regular assessment of dialysis patients and measured every 1 to 3 months. PMID- 12553437 TI - Protein status in pancreatitis--transthyretin is a sensitive biomarker of malnutrition in acute and chronic pancreatitis. AB - Malnutrition may develop in acute pancreatitis (AP), accompanied by hypermetabolism and high nutritional requirements, and in chronic pancreatitis (CP). We measured the incidence of protein malnutrition in AP and CP by comparing different serum biomarkers of protein metabolism and inflammation. Thirty-five patients with acute (27 moderate, 8 severe), and 35 with chronic, pancreatitis were enrolled in the study. Serum transthyretin, albumin, transferrin and C reactive protein (CRP) concentrations were measured in AP at admission, after 1 and 2 weeks of jejunal feeding, and in patients with CP at follow-up. In AP, at admission the transthyretin level was low in 74%, transferrin in 48%, and albumin in 29% of patients. In severe pancreatitis, transthyretin levels were significantly lower than in moderate forms (7.5 +/- 2.43 vs. 14.39 +/- 6.8 mg/dl, p < 0.005). Transthyretin levels increased significantly after 2 weeks of jejunal feeding (p < 0.05). In CP, transthyretin levels were decreased in 37%, transferrin in 27%, and albumin in 12% of patients. We found significantly lower transthyretin levels in alcohol-related CPthan in other forms (18.5 +/- 8.3 vs. 30.2 +/- 5.7, p < 0.01). Transthyretin correlated positively with albumin and transferrin and negatively with CRP Transthyretin seems to be a sensitive biomarker of protein status and metabolic stress. Monitoring nutritional status through measurement of serum proteins is important for optimal treatment of AP and CP. PMID- 12553438 TI - Assessment of nutritional status in organ transplant: is transthyretin a reliable indicator? AB - Transthyretin has been proposed as a nutritional index to screen for malnutrition and monitor the metabolic response to dietary intervention. In the presence of inflammation, circulating transthyretin levels drop regardless of optimal caloric intake. In this case, due to its rapid turnover, the pattern of transthyretin, monitored by means of repeated measures, could indicate the metabolic status (catabolism vs. anabolism). The aim of this review is to investigate the possible role of transthyretin as a nutritional parameter in organ transplantation. The literature on nutritional assessment in transplantation was reviewed and all the data regarding circulating transthyretin levels were analyzed. It appears that, on the one hand, the transthyretin level reflects closely dietary manipulations; on the other hand, it is affected by the inflammatory status. Consequently, interpretation could be difficult during the acute phase immediately after the transplant. Moreover, the role of transthyretin in monitoring the hepatic synthetic function in liver transplant is discussed. In conclusion, transthyretin is a reliable indicator of nutritional status in transplant candidates and potentially useful in the post-transplant phase if the inflammatory status is taken into account. PMID- 12553439 TI - Body composition and nutritional parameters in HIV and AIDS patients. AB - Undernutrition is a frequent complication of evolutive and chronic HIV (human immunodeficiency virus) infection characterized by bodyweight loss and changes in body composition. The Centers for Disease Control and Prevention define AIDS wasting as involuntary loss of more than 10% of body weight, plus more than 30 days of either diarrhea, or weakness and fever. Wasting syndrome has been considered as a case definition of the AIDS disease since 1987. Wasting syndrome is clearly linked to disease progression and death. Despite the progress under the era of highly active antiretroviral therapy (HAART), wasting is still a problem for people with AIDS. A small part of the weight lost is fat. More important is the loss of "lean body mass", which is mostly muscle. Body composition changes during HIV infection are different from those observed in food deprivation. Under the era of HAART, a HIV-associated adipose redistribution syndrome (HARS) was described that associates subcutaneous lipoatrophy and abdominal obesity linked to various metabolic disorders. Several factors contribute to wasting syndrome. Not only low food intake and poor nutrient absorption, but mainly altered metabolism (increased resting energy expenditure) and specific disturbances in protein turnover, which is also increased. Nutritional evaluation of HIV-infected patients should include the measurement of body composition and analysis of nutritional parameters, including albumin, transthyretin and C-reactive protein. Transthyretin seems to be particularly useful to follow the recovery period of malnutrition. PMID- 12553440 TI - C-reactive protein to transthyretin ratio for the early diagnosis and follow-up of postoperative infection. AB - The clinical usefulness of C-reactive protein (CRP) and of transthyretin (TTR) for the early diagnosis and follow-up of infection after an open fracture was prospectively investigated (cohort A). It was complemented by a retrospective study of trauma patients admitted to an intensive care unit (cohort B). Serial determinations of serum CRP and TTR concentrations were first performed in uninfected patients from cohort A to define a reference profile during the early postoperative period. It showed a concomitant increase in CRP and decrease in TTR concentrations, followed by progressive return to initial values in patients free from bacterial infection. Variations of the CRP/TTR ratio were analyzed. Recovery phase was defined by an exponential evolution of the two plasma proteins and of their ratio value. The CRP and TTR concentrations were independent of sex and severity of the trauma. In the case of postoperative infection, patients of cohort A revealed amplified CRP and TTR responses usually preceding the occurrence of clinical signs. During successful antibiotic therapy, their recovery response became superimposable to that of the reference group. The same profiles were recorded in cohort B patients admitted with lower limb fractures or various types of trauma. This suggests that observations made on cohort A can be extrapolated to othertrauma patients. We recommend that serial measurements of CRP and TTR and of their ratio should be performed every 2 days to appropriately follow-up these patients. PMID- 12553441 TI - The prognostic value of nutritional and inflammatory indices in critically ill patients with acute respiratory failure. AB - In critically ill patients suffering from acute respiratory failure, weaning from ventilatory assistance is a key survival factor in intensive care units (ICU). The aim of this study was to provide deeper insight into laboratory methods allowing improved monitoring of that critical period. Eighty-three ICU patients (mean age 63.9 years), classified according to the Second Acute Physiology and Chronic Health Evaluation criteria, were submitted to mechanical ventilation, antibiotherapy and nutritional support. Weaning attempts required degressive pressure support ventilation. The biological status of the patients was assessed by the serial measurement of inflammatory (C-reactive protein and alpha1-acid glycoprotein) and of nutritional (albumin and transthyretin) indicators whose aggregation yields a prognostic inflammatory and nutritional index (PINI). Statistical analyses compared ventilatory and biological data recorded on admission and at the time of extubation. Results showed that vital capacity and plasma concentrations of albumin and transthyretin rose, whereas rapid shallow breathing index, C-reactive protein and PINI values declined during the tested period. Persistent low transthyretin concentrations were predictive of lethality while increased values were associated with improved ventilatory performances. The PINI scoring formula worked as an independent predictor of the weaning trial outcome. The study underlined the value of the PINI system for the successful management of the weaning procedure. PMID- 12553442 TI - Transthyretin: its response to malnutrition and stress injury. clinical usefulness and economic implications. AB - Serum transthyretin is an ideal marker for monitoring patients who are malnourished or have metabolic consequences of acute stress injury because it has a short half-life, it measures the level of metabolic deficit, the response to nutritional metabolic support, and because it is a prognostic indicator. Mounting clinical evidence indicates that the use of transthyretin to assess and monitor a patient's nutritional status results in improved treatment outcomes and lower overall healthcare costs. PMID- 12553443 TI - Outcomes of continuous process improvement of a nutritional care program incorporating TTR measurement. AB - Early assessment of protein calorie malnutrition (PCM) can improve the outcome for hospitalized patients by allowing the initiation of nutrition support if required. In addition, monitoring nutritional status during the hospital stay can identify a decline in or improvement of PCM so that alterations to treatment regimens can be made if needed. The visceral protein albumin is the traditional laboratory indicator of PCM. In the past decade another protein has been lauded as a superior marker that can be used in conjunction. We undertook several studies to test the effectiveness of TTR as an aid in nutritional assessment. We found TTR to be a sensitive measure of nutritional status, allowing for earlier assessment and intervention, thus reducing length of stay and other hospital associated costs. Based on these findings, our hospital generated and implemented a multidisciplinary nutrition care program. Transthyretin is an integral portion of this program; levels are determined on admission and repeated twice weekly until discharge. PMID- 12553444 TI - The role of visceral protein markers in protein calorie malnutrition. AB - Despite substantial evidence of the crucial role protein calorie malnutrition (PCM) plays in the occurrence of complications, increased length of stay, and cost of care in hospitalized populations, no standard approach for screening and monitoring the nutritional status of patients initially and throughout admission currently exists. Recognizing that there is a growing public and professional recognition of the importance of malnutrition, a large patient population (30 55%) at risk for PCM, and an even larger population experiencing declining nutritional status during hospitalization, this study examined the feasibility of a full-scale study to assess the value of two biochemical markers, transthyretin and albumin, for detecting and monitoring PCM in hospitalized patients. It was demonstrated that these two markers do provide important information predictive of outcomes for those they identify at risk for PCM. The patients who entered the study with or developed low transthyretin and albumin experienced poorer health outcomes and higher costs of care. Their discharge occurred in an early phase of recovery, with significant implications for after-discharge care. The full-scale study must consider severity of illness and other confounders during randomization and, preferably, be conducted in institutions that currently do not use transthyretin for nutrition assessment. PMID- 12553445 TI - Influence of storage temperature on infectious hematopoietic necrosis virus detection by cell culture isolation and RT-PCR methods. AB - The detection of infectious hematopoietic necrosis virus (IHNV) in infected rainbow trout Oncorhynchus mykiss and in cell culture supernatants stored under different conditions was studied. IHNV-positive fish visceral organ homogenates and cell culture supernatants were incubated at 4 and 25 degrees C. Virus titre was measured by virus isolation on epithelioma papulosum cyprini (EPC) cells and the IHNV RNA was detected by RT-PCR and semi-nested RT-PCR. The influence of repeated freezing and thawing on the virus isolation from organ homogenates and from cell culture supernatants was studied as well. It was possible to isolate the virus from IHNV-positive organ material during the 3 d of incubation at 4 degrees C but, only on the first day of incubation at 25 degrees C. Viral RNA could be amplified during the incubation period of 35 d at 4 degrees C but only during 8 d of incubation at 25 degrees C. In IHNV-infected cell culture supernatant stored at 4 degrees C, it was possible to detect virus for 36 and 16 d in supernatant stored at 25 degrees C. Viral RNA could be followed by using molecular methods during the entire experimental period of 123 d. Each cycle of freezing and thawing of samples resulted in a reduction of IHNV titre in the suspension of visceral organs, while the virus titre in cell culture supernatant remained almost the same following 33 freezing-thawing cycles. The present results show that rapid laboratory processing and storage of potentially virus containing tissue samples as well as the use of different detection methods are very important for efficient IHNV diagnosis. PMID- 12553446 TI - Epizootic cutaneous papillomatosis in roach Rutilus rutilus: sex and size dependence, seasonal occurrence and between-population differences. AB - Epidemiology of epidermal papillomatosis in roach Rutilus rutilus was studied in 11 roach populations in Finland in 1999 and 2000 during the spawning period. In addition, the seasonal pattern of the disease was described in 1 population. Papilloma tumours were observed on the skin and fins of roach in 8 populations out of 11. Prevalence of papillomatosis varied from 0 to 62%, being higher in populations subject to industrial or sewage effluents. Males and large fish had higher prevalence of papilloma tumours. The disease outbreak peaked during the spawning period and signs of recovered skin on fish were observed later during the year. The gender dependence in the disease prevalence found in the study may indicate differences in stress and immunocompetence between roach males and females during the spawning period. PMID- 12553447 TI - Exocrine pancreatic carcinogenesis in the guppy Poecilia reticulata. AB - Exocrine pancreatic neoplasms developed in the guppy Poecilia reticulata following exposure to the direct-acting carcinogen methylazoxymethanol acetate (MAM-Ac). Fish 6 to 10 d old were exposed to nominal, non-toxic concentrations of 4 and 10 mg MAM-Ac l(-1) for 2 h and then transferred to carcinogen-free water for grow-out. Whole specimens were sampled monthly up to 9 mo post-exposure to follow the histologic progression of the lesions. No neoplasms occurred in 119 control specimens examined. Pancreatic acinar cell adenomas and carcinomas occurred in 42 of 243 (17%) of the specimens exposed to MAM-Ac. As in earlier studies, specimens exposed to the low MAM-Ac concentration exhibited a higher pancreatic neoplasm incidence (27.8%) than those exposed to the high concentration (7.8%). Acinar cell adenomas accounted for 27 of the 42 neoplasms. Adenomas exhibited a high degree of acinar cell differentiation and some contained foci of atypical acinar cells that were less differentiated and more basophilic than were surrounding adenoma cells. Carcinomas occurred in 15 specimens and exhibited a range of cellular patterns. Although no distant metastases were found, carcinomas tended to invade neighboring tissues and organs. The occurrence of carcinogen-induced pancreatic neoplasms in guppies strengthens the usefulness of small fish species in carcinogen testing and provides an additional model for studying pancreatic neoplasia. PMID- 12553448 TI - Infection of barramundi Lates calcarifer with Streptococcus iniae: effects of different routes of exposure. AB - The use of various challenge techniques has allowed the formation of a hypothesis for the mode of infection of Streptococcus iniae in barramundi. A bacterial dose of 1 x 10(3) colony forming units (cfu), corresponding to the LD50, delivered orally to barramundi could initiate the sub-acute form of the disease observed at the farms. The acute form of the disease could be initiated through bath exposure to the pathogen. S. iniae was equally as infective in freshwater, saltwater or when fish were subject to skin trauma prior to exposure, with LD50 values of 3.2 x 10(4), 2.0 x 10(4), 3.2 x 10(4) cfu, respectively, when observed over a 10 d period. It is suggested that sub-acute infection occurs orally, with mass mortalities occurring through the increased presence of the bacterium in the environment. PMID- 12553449 TI - Host site of activity and cytological effects of histone-like proteins on the parasitic dinoflagellate Amyloodinium ocellatum. AB - Histone-like proteins (HLPs) are broad-spectrum, endogenously produced antibiotics which we have isolated from tissues of rainbow trout Oncorhynchus mykiss and hybrid striped bass (Morone saxatilis male x M. chrysops female). Here, we show that HLP-1, which has high sequence homology to histone H2B, equally inhibited both young and mature trophonts of the important ectoparasite Amyloodinium ocellatum. In addition to direct killing of Amyloodinium trophonts, there was evidence that HLP-1 from both rainbow trout and hybrid striped bass caused severe developmental abnormalities, including delayed development, in both the parasitic trophont stage as well as the reproductive tomont stage. The deleterious effects of HLP-1 also were manifested in what appeared to be 'delayed mortality', where parasites of normal appearance would die later in development. Similar serious damage was also seen with calf histone H2B and the unrelated peptide antibiotic magainin 2. A comparison of the antibiotic activity in mucus versus epidermis compartments of the skin of hybrid striped bass suggested that the majority of antibiotic (including HLP-1) activity resided in the epidermis, although some activity was present in the mucus. These data suggest that normal, nonimmune fish skin contains potent defenses against protozoan ectoparasites and that the effects of these defenses may extend beyond their transient interactions with the parasites, which has important implications for this host-parasite relationship. PMID- 12553450 TI - Continuous in vitro culture of the carpet shell clam Tapes decussatus protozoan parasite Perkinsus atlanticus. AB - Continuous in vitro cultures of the clam Tapes decussatus parasite Perkinsus atlanticus were established from infected gill fragments, infected haemolymph and parasite hypnospores isolated from infected gill fragments following incubation in Ray's fluid thioglycollate medium (RFTM). No continuous cultures could be initiated from P. atlanticus zoospores. Cultures initiated from hypnospores yielded the highest percentage of continuous cultures (100%, 6/6), followed by cultures initiated from gill fragments (93%, 43/46) and from haemolymph (30%, 3/10). Failures to establish continuous cultures were due to microbial contamination. The source of parasite influenced the success rate, the time taken to establish cultures and the size of cultured cells. In vitro proliferation of parasite cells was mainly by vegetative multiplication. Zoosporulation, yielding motile biflagellated zoospores, was observed at a low frequency (< 1% of dividing cells) in every culture. Morphology of cultured cells examined with light and transmission electron microscopy corresponded to that of P. atlanticus found in clam tissues. Cultured cells enlarged in RFTM and stained blue-black with Lugol's solution, which are characteristics of the Perkinsus species cells. DNA sequences of the internal transcribed spacer (ITS) region of the ribosomal RNA gene complex matched those of P. atlanticus. All cultures were established in a medium designated JL-ODRP-2A that was similar in composition to the culture medium JL ODRP-1 originally used to propagate Perkinsus marinus in vitro. Proliferation of P. atlanticus in vitro could be supported by the commercial culture medium (1:2 v/v) DME:Ham's F-12 with fetuin. PMID- 12553451 TI - Molecular diagnostics, field validation, and phylogenetic analysis of Quahog Parasite Unknown (QPX), a pathogen of the hard clam Mercenaria mercenaria. AB - Quahog Parasite Unknown (QPX) is a protistan parasite that causes disease and mortality in the hard clam Mercenaria mercenaria. PCR primers and DNA oligonucleotide probes were designed and evaluated for sensitivity and specificity for the QPX organism specifically and for the phylum Labyrinthulomycota in general. The best performing QPX-specific primer pair amplified a 665 bp region of the QPX small-subunit ribosomal DNA (SSU rDNA) and detected as little as 1 fg cloned QPX SSU rDNA and 20 fg QPX genomic DNA. The primers did not amplify DNA of uninfected hard clams M. mercenaria or of the thraustochytrids Schizochytrium aggregatum, Thraustochytrium aureum, and T. striatum. The general labyrinthulomycete primers, which were designed to offer broader specificity than the QPX primers, amplified a 435 bp region of SSU rDNA from QPX, and a 436 to 437 bp region of SSU rDNA from S. aggregatum, T. aureum, and T. striatum, but did not amplify that of the clam M. mercenaria. Field validation of the QPX-specific primer pair, through comparative sampling of 224 clams collected over a 16 mo period from a QPX endemic site in Virginia, USA, indicated that the PCR assay is equivalent to histological diagnosis if initially negative PCR products are reamplified. Oligonucleotide DNA probes specific for QPX and the phylum Labyrinthulomycota were evaluated for in situ hybridization assays of cell smears or paraffin-embedded tissues. Two DNA probes for QPX offered limited sensitivity when used independently; however, when used together as a probe cocktail, sensitivity was greatly enhanced. The probe cocktail hybridized to putative QPX organisms in tissues of hard clams collected from Virginia, New Jersey, Massachusetts and Canada, suggesting that the QPX organisms in these areas are either very closely related or the same species. The QPX probe cocktail did not hybridize with clam tissue or with the thraustochytrids S. aggregatum, T. aureum, and T. striatum. The labyrinthulomycete DNA probe hybridized with QPX and the 3 thraustochytrids, with no background hybridization to clam tissue. SSU rDNA sequences were obtained for the putative QPX organisms from geographically distinct sites. Phylogenetic analyses based on the QPX and Labyrinthulomycota sequences confirmed earlier reports that QPX is a member of this phylum, but could not definitively demonstrate that all of the QPX organisms were the same species. PMID- 12553452 TI - White spot syndrome virus (WSSV) infects specific hemocytes of the shrimp Penaeus merguiensis. AB - White spot syndrome virus (WSSV) was specifically detected by PCR in Penaeus merguiensis hemocytes, hemolymph and plasma. This suggested a close association between the shrimp hemolymph and the virus. Three types of hemocyte from shrimp were isolated using flow cytometry. Dynamic changes of the hemocyte subpopulations in P. merguiensis at different times after infection were observed, indicating that the WSSV infection selectively affected specific subpopulations. Immunofluorescence assay (IFA) and a Wright-Giemsa double staining study of hemocyte types further confirmed the cellular localization of the virus in the infected hemocytes. Electron microscopy revealed virus particles in both vacuoles and the nucleus of the semigranular cells (SGC), as well as in the vacuoles of the granular cells (GC). However, no virus could be detected in the hyaline cells (HC). Our results suggest that the virus infects 2 types of shrimp hemocytes--GCs and SGCs. The SGC type contains higher virus loads and exhibits faster infection rates, and is apparently more susceptible to WSSV infection. PMID- 12553453 TI - Spring viremia of carp (SVC). AB - Spring viremia of carp (SVC) is an important disease affecting cyprinids, mainly common carp Cyprinus carpio. The disease is widespread in European carp culture, where it causes significant morbidity and mortality. Designated a notifiable disease by the Office International des Epizooties, SVC is caused by a rhabdovirus, spring viremia of carp virus (SVCV). Affected fish show destruction of tissues in the kidney, spleen and liver, leading to hemorrhage, loss of water salt balance and impairment of immune response. High mortality occurs at water temperatures of 10 to 17 degrees C, typically in spring. At higher temperatures, infected carp develop humoral antibodies that can neutralize the spread of virus and such carp are protected against re-infection by solid immunity. The virus is shed mostly with the feces and urine of clinically infected fish and by carriers. Waterborne transmission is believed to be the primary route of infection, but bloodsucking parasites like leeches and the carp louse may serve as mechanical vectors of SVCV. The genome of SVCV is composed of a single molecule of linear, negative-sense, single-stranded RNA containing 5 genes in the order 3'-NPMGL-5' coding for the viral nucleoprotein, phosphoprotein, matrix protein, glycoprotein, and polymerase, respectively. Polyacrylamide gel electrophoresis of the viral proteins, and sequence homologies between the genes and gene junctions of SVCV and vesicular stomatitis viruses, have led to the placement of the virus as a tentative member of the genus Vesiculovirus in the family Rhabdoviridae. These methods also revealed that SVCV is not related to fish rhabdoviruses of the genus Novirhabdovirus. In vitro replication of SVCV takes place in the cytoplasm of cultured cells of fish, bird and mammalian origin at temperatures of 4 to 31 degrees C, with an optimum of about 20 degrees C. Spring viremia of carp can be diagnosed by clinical signs, isolation of virus in cell culture and molecular methods. Antibodies directed against SVCV react with the homologous virus in serum neutralization, immunofluorescence, immunoperoxidase, or enzyme-linked immunosorbent assays, but they cross-react to various degrees with the pike fry rhabdovirus (PFR), suggesting the 2 viruses are closely related. However, SVCV and PFR can be distinguished by certain serological tests and molecular methods such as the ribonuclease protection assay. PMID- 12553454 TI - Lateral thinking: a different way of benefiting from reading equine veterinary journal and other scientific publications. PMID- 12553455 TI - The neutrophil: understanding ancient and powerful responses in the inflammatory balance. PMID- 12553456 TI - Osteochondrosis and more ideas on an enzymic pathogenesis. PMID- 12553457 TI - Needle electromyography in the horse compared with its principles in man: a review. PMID- 12553458 TI - Magnetic resonance imaging of the equine foot: 15 horses. AB - REASONS FOR PERFORMING STUDY: Foot pain is a common cause of equine lameness and there have been significant limitations of the methods available for the diagnosis of the causes of foot pain (radiography, nuclear scintigraphy and ultrasonography). Until recently, magnetic resonance imaging (MRI) in the horse has been limited to examination of cadaver limbs. OBJECTIVES: Our purpose was to 1) describe MRI of the foot in live horses, 2) describe MRI findings in horses with foot pain in which a definitive diagnosis could not be established by alternative means and 3) correlate MRI findings with other methods of clinical investigation. METHODS: The feet of 15 horses with unilateral (12) or bilateral (3), forelimb (14) or hindlimb (1) lameness associated with foot pain of previously ill-defined origin were examined using MRI. The horses were examined in right lateral recumbency under general anaesthesia, with the feet positioned in the isocentre of a flared end 1.5 Tesla GE Signa Echospeed magnet. Images were obtained in sagittal, transverse and dorsal planes using 3-dimensional (3D) T2* gradient echo (GRE), spoiled gradient echo, fat-saturated 3D T2* GRE and short inversion recovery sequences. Image acquisition took approximately 1 h. RESULTS: Abnormalities of the distal interphalangeal joint (DIP) cartilage and/or subchondral bone, periarticular osteophyte formation, distension of the DIP joint capsule with or without synovial proliferation, distension of the navicular bursa with or without evidence of chronic inflammation, surface and core lesions in the deep digital flexor tendon, abnormal signal within the navicular bone, evidence of mineralised fragments in the distal sesamoidean impar ligament, irregular outline of and signal in the medial cortex of the distal phalanx, and an abnormal signal on the dorsal aspect of the distal phalanx consistent with laminitis were identified. CONCLUSIONS: MRI permits the diagnosis of a variety of lesions involving different structures within the foot that cannot be diagnosed using other means, thus enhancing our knowledge of the causes of foot pain. POTENTIAL RELEVANCE: With further experience it is likely that lesions involving other structures will also be identified. Long-term follow-up data is required to determine the prognosis for the injuries described. PMID- 12553459 TI - Validation of the 13C-octanoic acid breath test for measurement of equine gastric emptying rate of solids using radioscintigraphy. AB - REASONS FOR PERFORMING STUDY: Disordered gastric motility may be a significant factor in the pathogenesis of many equine conditions. Although tests for liquid phase emptying rate have been validated in the horse, there are no effective tests for solid phase emptying measurement that can be performed routinely in the field. OBJECTIVES: The objective of this study was the assessment of a novel stable isotope technique, the 13C-octane acid breath test (13C-OABT), for the measurement of gastric emptying of solid ingesta, by direct comparison with the optimum method of gastric scintigraphy. METHODS: To facilitate dual measurement of gastric emptying, a test meal was used containing baked egg yolk labelled with both 13C-octanoic acid and 99mtechnetium sulphur colloid. Simultaneous, serial lateral gastric scintigraphs and expiratory breath samples were obtained in 12 healthy horses after voluntary ingestion of the test meal. Analysis of breath 13CO2:12CO2 ratio was performed by continuous flow isotope ratio mass spectrometry. Power regression was used to determine the gastric emptying coefficient, the gastric half-emptying time (t 1/2) and duration of the lag phase (tlag). RESULTS: Significant correlations (P < 0.001) were found between the 2 techniques for measurement of both t 1/2 and tlag. In addition, scintigraphic left t 1/2 was correlated significantly to breath test gastric emptying coefficient (P < 0.001). CONCLUSIONS: It was concluded that the 13C-octanoic acid breath test is a reliable diagnostic procedure to measure gastric emptying rate of solids in the horse. POTENTIAL RELEVANCE: Being safe, noninvasive and easy to perform, this test has potential value as both sensitive diagnostic modality and humane research tool for motility studies. PMID- 12553460 TI - Potentiation of the extracellular release of equine neutrophil elastase and alpha 1-proteinase inhibitor by a combination of two bacterial cell wall components: fMLP and LPS. AB - REASONS FOR PERFORMING STUDY: Lipopolysaccharide (LPS) and N-formyl-methionyl leucyl-phenylalanine (fMLP)-like peptides are Gram-negative bacterial cell wall components which, when released into the peripheral circulation in endotoxaemia, have the potential to activate leucocytes. In vitro, equine neutrophils require priming with LPS in order to generate reactive oxygen intermediates (ROI) in response to fMLP. OBJECTIVES: The aim of this study was to examine whether the release of other neutrophil products is similarly dependent on prior priming with LPS. In particular, neutrophil elastase (NE), a potent proteolytic enzyme, and its major inhibitor, alpha-1 proteinase inhibitor, were investigated. METHODS: Neutrophils were isolated from equine peripheral blood (n = 5) by discontinuous Percoll gradient preparative centrifugation and primed with LPS prior to stimulation with fMLP. ROI were measured by lucigenin dependent chemiluminescence (LDCL). Concentrations of NE and API were determined by ELISA on cell free supernatants taken at 0, 2, 10, 30, 60 and 90 mins post stimulus. Data was analysed by Kruskal-Wallis and Mann-Whitney Tests. RESULTS: Sequential exposure of Percoll purified equine blood neutrophils in vitro to LPS followed by fMLP resulted in the greatest release of NE from equine neutrophils and was required for ROI generation. However, LPS or fMLP stimulation alone resulted in an increase in NE release compared to unstimulated control cells. In contrast, significant API release was only induced by LPS stimulation or fMLP stimulation only after LPS priming, not fMLP on its own. CONCLUSIONS: These results suggest that different stimuli (fMLP or LPS) are capable of invoking similar responses from equine neutrophils with respect to NE release yet different ones with respect to API release. POTENTIAL RELEVANCE: In addition, demonstration of elastase release induced by LPS and/or fMLP suggests that monitoring serum elastase levels is a potential diagnostic tool for detecting the early onset of endotoxaemia in the horse. PMID- 12553461 TI - Differential distribution of cathepsins B and L in articular cartilage during skeletal development in the horse. AB - REASONS FOR PERFORMING STUDY: This study was designed to examine a new role for cysteine proteinases in the process of endochondral ossification. OBJECTIVES: The aim of the present study was to investigate the presence and distribution of cathepsin B and cathepsin L in equine articular cartilage during development. METHODS: Full-depth cartilage samples from a total of 40 horses (age range: 4 month fetuses to 2 years) were examined and enzymes detected by immunocytochemical localisation. RESULTS: Observations on the presence of cathepsins B and L revealed significant age-related differences, resulting in clear division of the animals into 2 age groups: i) fetuses and neonates; ii) young growing horses (age 4 weeks to 2 years). Cathepsin B was not detected in cartilage from the majority of fetuses and neonates but was located characteristically in chondrocytes at the articular surface and hypertrophic zone in all growing horses. In contrast, cathepsin L was predominantly present in fetal and neonatal cartilage, located primarily in proliferating chondrocytes. CONCLUSIONS: This study is the first to demonstrate differential and site specific roles for cathepsin B and cathepsin L in skeletal development in the horse. POTENTIAL RELEVANCE: The demonstrated involvement of cathepsins B and L in endochondral ossification is of relevance to developmental orthopaedic diseases such as osteochondrosis in which there is a focal failure of bone formation. PMID- 12553462 TI - Ultrasonographic abnormalities detected in the sacroiliac area in twenty cases of upper hindlimb lameness. AB - REASONS FOR PERFORMING STUDY: The pelvis is covered with extensive musculature and often presents a challenge in diagnostic imaging. Ultrasonography provides diagnostic information about soft tissue, articular cartilage and bone surfaces, although little information exists about the normal ultrasonographic appearance of the equine sacroiliac region. OBJECTIVES: To determine the clinical applicability of ultrasonographic examination in horses with sacroiliac pain. METHODS: Horses presented to the University of Minnesota Veterinary Teaching Hospital for hindlimb lameness were evaluated and lower limb lameness was ruled out with examination and local anaesthesia. Twenty cases were diagnosed with sacroiliac pain, characterised by response to palpation over the tuber sacrale. Seven of the 20 had visible asymmetry of the tuber sacrale. Ultrasonography was performed percutaneously and per rectum. Structures imaged were measured and compared to normal reference measurements. Other methods used to confirm diagnoses included thermography, scintigraphy and radiology. RESULTS: All 20 cases with sacroiliac pain showed ultrasonographic abnormalities of the sacroiliac area. Abnormalities were detected in the dorsal sacroiliac ligaments. Ligament size was significantly different from the normal ranges (P < 0.01). Malalignment of the sacroiliac area and sacral fractures were also imaged. CONCLUSIONS: Ultrasonography of the sacroiliac region aided diagnosis of ligament damage and displacement of bone surfaces; and may aid in monitoring the progression of healing. POTENTIAL RELEVANCE: Diagnostic ultrasound is readily available to most practitioners. Therefore the findings of this study show that diagnosis of sacroiliac injuries is possible in the field. PMID- 12553463 TI - Estimates of heritability for ossification of the cartilages of the front feet in the Finnhorse. AB - REASONS FOR PERFORMING STUDY: Ossification of the cartilages in the foot is common in coldblooded horses, but prevalence as well as ossification pattern varies between breeds; research on estimation of heritability for the condition has been limited. OBJECTIVES: Our purpose was to calculate estimates of 1) heritability for ossification parameters of the cartilages in the front feet of the Finnhorse and 2) genetic correlations between the different ossification parameters. METHODS: Estimates of heritability for different ossification parameters of the cartilages in the front feet and of genetic correlations between different parameters were evaluated in data consisting of dorsopalmar radiographs of the front feet and pedigree up to the fourth generation of 964 Finnhorses (age > or = 2 years; 345 females and 619 males). Ossification at the base of the cartilages and total ossification, including separate centres of ossification, were graded 0-5 according to the most proximal point of ossification and the presence of separate centres of ossification was recorded separately. The data were analysed using a model including a fixed sex-age subclass effect and a random animal effect. Variance and covariance components were estimated with the restricted maximum likelihood (REML) method assuming an animal model. RESULTS: Females had more ossification than males and this difference increased with age. The ossification in the medial cartilages progressed significantly with age in females but not in males. In the lateral cartilages, the increase in ossification with age was evident in both sexes. Separate centres of ossification were an incidental finding. Estimates of heritability for the ossification parameters varied slightly between the cartilages as well as between the front feet. For total ossification, the estimates of heritability varied between 0.31 (right lateral) and 0.50 (left medial) and were slightly higher for ossification at the base of the cartilages. High genetic correlations were found for total ossification between the medial and lateral cartilages in the foot and for parallel ossification parameters between the right and left foot. CONCLUSIONS: The estimates of heritability were relatively high, and both cartilages and front feet have a similar genetic tendency to ossify. POTENTIAL RELEVANCE: This information is useful in further studies assessing other factors contributing to the ossification process and also in the planning and modifying breeding programmes. PMID- 12553464 TI - Changes in nasal mucosal innervation in horses with grass sickness. AB - REASONS FOR PERFORMING STUDY: Equine grass sickness is a dysautonomia characterised by widespread destruction of autonomic ganglia, resulting in the clinical signs of dysphagia, constipation, profuse sweating, tachycardia, rhinitis sicca and high mortality rate. Rhinitis sicca is a common finding in horses with the chronic form and we have postulated that alterations in autonomic innervation of the nasal mucosa might underlie this clinical presentation. OBJECTIVES: In this study, the expression and distribution of nerve fibres immunoreactive for calcitonin gene-related peptide (CGRP), substance P (SP), the general neuronal marker protein gene-product 9.5 (PGP 9.5; ubiquitin) and the intermediate neurofilaments (PAN-N; neurorfilaments L, M and H) in the nasal mucosa of normal horses (n = 10) and horses with EGS (n = 18; acute n = 8, subacute n = 3, chronic n = 7) was assessed. METHODS: Innervation density and distribution was investigated in the different groups using standard immunohistochemical techniques. RESULTS: Significant differences were noted when comparing the density and distribution of nerve fibres immunoreactive for PGP 95 and PAN-N, with PGP 95 consistently giving better staining in all groups and at all sites in the nasal mucosa. An apparent increase in the density of innervation was noted for acute vs. normal cases. A significant reduction in the density of innervation was noted only with PAN-N when comparing normal horses and acute cases with the chronic group (P < 0.05). CGRP and SP immunoreactive nerve fibres were typically most abundant in the epithelial and subepithelial layers, but the quality of staining and nerve fibre density was greater for SP, achieving significant difference in several comparisons. The density of innervation for SP was significantly reduced in the chronic group compared to the normal and acute groups (P < 0.01). A significant decrease was noted for CGRP only for the acute and chronic groups (P<0.05). CONCLUSIONS: These results demonstrate a reduction in the expression of the sensory neuropeptides in nasal mucosal innervation as a consequence of equine dysautonomia, and may underlie the clinical presentation of rhinitis sicca noted with this disease. POTENTIAL RELEVANCE: Nasal biopsy may be of use in antemortem diagnosis of grass sickness and identification of mucosal denervation; and might also be useful in the treatment of rhinitis in EGS cases. PMID- 12553465 TI - Influence of liver copper status of mare and newborn foal on the development of osteochondrotic lesions. AB - REASON FOR PERFORMING STUDY: To elucidate the highly contentious role of copper in the pathogenesis of osteochondrosis. HYPOTHESIS: There would be no relationship between liver copper concentration of mares and foals and incidence of radiographically detectable osteochondrotic lesions in foals and yearlings was tested. METHODS: Liver copper concentration was assessed in biopsies taken within 4 days after birth from both mares and foals and from the same foals at age 5 months. Biopsies were taken in the standing, sedated animal under ultrasonographic guidance. Radiographs were taken of both hocks (lateromedial, dorsoplantar and dorsomedial-plantarolateral oblique views) and stifles (lateromedial and caudolateral-craniomedial oblique views) at ages 5 and 11 months and scored for the presence and severity of osteochondrotic lesions. RESULTS: Copper concentrations in newborn foals were high with a large variation (351 +/- 201 mg/kg DM). They declined until reaching values comparable to those in mature animals at 5 months (20 +/- 8 mg/kg DM; mares: 19 +/- 20 mg/kg DM). Radiographic osteochondrotic lesions decreased in number and severity from 5 to 11 months. This pattern was more predominant in the stifle than in the hock, as has been described previously. CONCLUSIONS: There was no relationship between foal or mare liver copper concentration and osteochondrosis status at either 5 or 11 months. However, osteochondrotic lesions in foals with low-level copper status at birth decreased significantly less in number and severity than those in foals with high-level copper status at birth. POTENTIAL RELEVANCE: It is concluded that copper is not likely to be an important factor in the aetiopathogenesis of osteochondrosis, but this study indicates that there may be a significant effect of high copper status on the natural process of repair of early lesions. PMID- 12553466 TI - Occurrence of infectious upper respiratory tract disease and response to vaccination in horses on six sentinel premises in northern Colorado. AB - REASONS FOR PERFORMING STUDY: Horses vaccinated against common agents of infectious upper respiratory disease (IURD) may not have detectable serum antibody and may not be protected from clinical disease. OBJECTIVES: The objectives of this study were to 1) investigate the serological response of horses to vaccination against influenza virus (H3N8 and H7N7) and equine herpesviruses (EHV) in a field setting and 2) evaluate associations among vaccination status, serum antibody concentrations, and occurrences of IURD in monitored horses. METHODS: In this study, horses on 6 Colorado premises were vaccinated parenterally against influenza virus and EHV, and serological response evaluated. Horses were monitored, and biological samples collected from individuals with clinical IURD and control horses. RESULTS: Of 173 horses, 61 (35.3%), 21 (12.1%) and 4 (2.3%) seroconverted in response to vaccination against EHV, influenza virus H7N7 and influenza virus H3N8, respectively. CONCLUSIONS: Outbreaks of IURD in study horses were associated with influenza virus H3N8 and Streptococcus equi infection, and serological response to vaccination with conventional products was poor. POTENTIAL RELEVANCE: These results confirm that horses may not respond with detectable serological responses to conventional vaccination against common respiratory viruses and, therefore, suggest that alternate methods of protecting horses against common respiratory viruses should be sought. PMID- 12553467 TI - Thoracic trauma in foals: post mortem findings. AB - REASONS FOR PERFORMING STUDY: Thoracic trauma occurs in newborn foals and may cause associated clinical signs; this condition remains poorly documented. OBJECTIVES: The purpose of this study was to describe the pathological features of thoracic trauma in newborn foals presented for necropsy examination between 1990 and 2000. METHODS: Necropsy reports of foals with thoracic trauma from 1990 2000 were reviewed. Subject details, clinical signs, thoracic and abdominal lesions were noted and analysed statistically. RESULTS: Sixty-seven (9%) of 760 necropsied foals had thoracic trauma. In 19 foals, fractured ribs were considered to be the cause of death (Group A). The remaining foals had fractured ribs (Group B, n = 20) or rib contusions (Group C, n = 28) that were incidental findings. Ribs 3 to 8 accounted for 86% of the traumatised bones. The most common site of injury was the costochondral junction and an area immediately above it (94%). In Group A, all but 2 foals died within the first 8 days postpartum. Haemothorax and subsequent pulmonary collapse was cited most commonly as the cause of death (53%). Diaphragmatic rupture and hernia (n = 2) also occurred. CONCLUSIONS: The focal site, consistent location and presence of lesions during the first week post partum, all suggest that thoracic trauma in newborn foals probably occurs during parturition. POTENTIAL RELEVANCE: The description of lesions and site of occurrence of thoracic trauma in foals will increase awareness and improve the diagnosis and treatment of this life threatening condition. PMID- 12553468 TI - Dystocia in a referral hospital setting: approach and results. AB - REASONS FOR PERFORMING STUDY: Dystocia in the mare is an emergency in which duration has a profound effect on survival of the foal. Specific examination of the effects of dystocia duration on foal survival provides information to enable horse care personnel and veterinarians to manage these cases more effectively and maximise the chances of obtaining a live foal. HYPOTHESIS: Dystocia duration would have a negative impact on foal survival while method of dystocia resolution would not have an effect on foal survival. Additionally, we were interested in determining the effects of dystocia on subsequent fertility. METHODS AND RESULTS: In the years 1986-1999, 247 dystocias were admitted. Of these, 91 % resulted in survival and discharge of the mare, 42% in delivery of a live foal, and 29% of foals survived to discharge. Period from hospital arrival to delivery for foals alive at discharge (23.0 +/- 14.1 mins) was not significantly different than for foals not surviving (24.8 +/- 10.6 mins) (P > 0.05); and from chorioallantoic rupture to delivery for foals alive at discharge (71.7 +/- 343 mins) was significantly less than for foals not surviving (853 +/- 37.4 mins) (P < 0.05). Average predystocia live foaling rates for all mares with available records was 84%. Overall post dystocia live foaling rates over the entire period of this study were 67%. Of mares bred in the year of the dystocia, 59% had a live foal in the year following. CONCLUSIONS: Based on these results, dystocia duration has a significant effect on foal survival and resolution methods should be chosen to minimise this time, as the difference between mean dystocia duration for foals that lived and those that did not in this study was 13.6 mins. Post dystocia foaling rates reported here are higher than previously reported for both same season and overall breedings, indicating same-season breeding may be rewarding for select dystocia cases. POTENTIAL RELEVANCE: Dystocia resolution methods that minimise delivery time may maximise foal survival. Post dystoicia breeding may be rewarding in select cases. PMID- 12553469 TI - Clostridial myonecrosis in horses (37 cases 1985-2000). AB - REASONS FOR PERFORMING STUDY: Previous reports of clostridial myonecrosis have either focused on individual case reports or have been small retrospective studies reporting very high mortality rates. OBJECTIVES: The objective of this study was to describe the outcome of cases of clostridial myonecrosis submitted to 2 referral equine hospitals in the United States over a 15 year period. METHODS: A retrospective study of case material selected on the basis of positive Clostridium spp. culture or the identification of Clostridium spp. by specific fluorescent antibody testing from soft tissue wounds was performed at Cornell and Wisconsin. RESULTS: 37 cases of clostridial myonecrosis were documented. Twenty seven horses survived, 8 were subjected to euthanasia and 2 died during treatment for an overall survival rate of 73%. Twenty-five cases (68%) were associated with Clostridium perfringens alone, 6 cases (16%) with Cl. septicum alone, 4 cases with mixed clostridial infections (11%), 1 case with Cl. sporogenes and 1 with an unspeciated Clostridium spp. The highest survival rate of 81% was documented for those cases from which Cl. perfringens alone was isolated. The most common antecedent condition prior to referral was colic. The myonecrotic lesion occurred within 6-72 h of a soft tissue injection in 34 cases but was associated with a wound or laceration in the remaining 3 cases. Of the 34 cases associated with recent injections, 24 were associated with i.m. injections in the cervical region, 4 in the semimembranosus/semitendinosus region, 3 in the gluteal region, 2 with perivascular leakage of drugs administered into the jugular vein and 1 case developed simultaneously in the gluteal and neck region following injections at both sites. CONCLUSIONS: Clostridial myonecrosis can occur following the i.m. or inadvertent perivascular administration of a wide variety of commonly administered drugs. It is most common in the neck musculature. Aggressive treatment can be associated with survival rates of up to 81% for cases due to Cl. perfringens alone. Survival rates for other Clostridial spp. tend to be lower. POTENTIAL RELEVANCE: A combination of high dose i.v. antibiotic therapy and surgical fenestration/debridement is the best approach to cases of clostridial myonecrosis. With rapid diagnosis and therapeutic intervention, horses may have up to an 81% chance of survival. PMID- 12553470 TI - Effects of halothane anaesthesia on the cryopreservation of epididymal spermatozoa in pony stallions. PMID- 12553471 TI - Plasma angiotensin-converting enzyme (ACE) concentration in Thoroughbred racehorses. PMID- 12553472 TI - Isolation and implantation of autologous equine mesenchymal stem cells from bone marrow into the superficial digital flexor tendon as a potential novel treatment. PMID- 12553473 TI - Primary retroperitoneal tumour in a horse. PMID- 12553474 TI - Renal carcinoma with osseous and pulmonary metastases in an Arabian gelding. PMID- 12553475 TI - Rupture of the biceps tendon in a Thoroughbred steeplechase horse. PMID- 12553476 TI - Gene expression of genital human papillomaviruses and considerations on potential antiviral approaches. AB - Genital human papillomaviruses (HPVs) are carcinogenic to humans and are associated with most cases of cervical cancer, genital and laryngeal warts, and certain cutaneous neoplastic lesions. Five of the more than 50 known genital HPV types, HPV-6, -11, -16, -18 and -31, have become the models to study gene expression. The comparison of the studies of these five viruses and analyses of the genomic sequences of those genital HPV types that have not been transcriptionally studied make it likely that genital HPVs share most strategies for regulating their transcription. These strategies are quite different from those of unrelated human and animal papillomaviruses. Among these common properties are (i) a specific promoter structure allowing for fine-tuned negative feedback, (ii) a transcriptional enhancer that is specific for epithelial cells, (iii) regulation by progesterone and glucocorticoid hormones, (iv) silencers, whose principal function appears to be transcriptional repression in the basal layer of infected epithelia, (v) specifically positioned nucleosomes that mediate the functions of some enhancer and the silencer factors, (vi) nuclear matrix attachment regions that can, under different conditions, repress or stimulate transcription, and (vii) as yet poorly understood late promoters positioned very remote from the late genes. Most of these properties are controlled by cellular proteins that, due to their simultaneous importance for cellular processes, may not be useful as HPV-specific drug targets. It should be possible, however, to target complex cis-responsive elements unique to these HPV genomes by nucleotide sequence-specific molecules, such as antisense RNA, polyamides and artificial transcription factors. The application of small molecule-based drugs may be restricted to target proteins encoded by the HPV DNA, such as the replication factor E1 and the transcription/replication factor E2. PMID- 12553477 TI - Hyperlactataemia in HIV-infected patients: the role of NRTI-treatment. AB - BACKGROUND: Long-term treatment with nucleoside reverse transcriptase inhibitors (NRTIs) can induce mitochondrial dysfunction, most severely represented by lactic acidosis. Diagnostic tests for mitochondrial dysfunction are lacking, although persistently elevated serum lactate might be a surrogate marker. OBJECTIVES: To determine the occurrence of hyperlactataemia in HIV-infected patients on NRTI treatment and to evaluate the possible risk factors. METHODS: Cross-sectional analysis of lactic-acid levels in asymptomatic HIV-infected patients. Hyperlactactaemia was considered mild if between 2.0-5 mmol/l, serious if >5 mmol/l and lactic acidosis was defined as lactic acid levels >5 mmol/l with bicarbonate <20 mmol/l. Possible risk factors, such as current and preceding NRTI treatment as well as treatment with non-nucleoside reverse transcriptase inhibitors or protease inhibitors and concurrent liver disease, were analysed. RESULTS: Two hundred and twenty three asymptomatic HIV-infected patients were studied, including 174 patients (78%) on NRTI treatment, 12 patients (5%) treated without NRTIs and 37 patients (17%) not treated. Mild hyperlactataemia was found in 42 patients (19%), from whom 38/42 (90%) were NRTI-treated and the remaining patients (4/42, 10%) received no treatment (chi2, P<0.05). The significant risk factors for hyperlactataemia in the univariate analysis were NRTI-treatment as a group (P=0.03) and elevated ALT (P=0.008). In multivariate analysis NRTI use (P=0.05) and ALT level (P=0.03) remained a significant determinant of hyperlactataemia. Among the different individual NRTIs, a stavudine-containing (P=0.004) and a zalcitabine-containing (P=0.07) regimen were most notably associated with the development of hyperlactataemia, whereas for the combinations of NRTIs, such association was only found for stavudine/lamivudine (P=0.05). CONCLUSIONS: A correlation between hyperlactataemia and NRTI treatment was found, but the value of routine lactate measurement for individual treatment monitoring remains uncertain. PMID- 12553478 TI - Divergent distribution of HIV-1 drug-resistant variants on and off antiretroviral therapy. AB - OBJECTIVE: To investigate the distribution of drug-resistant HIV-1 variants in plasma RNA and peripheral blood monuclear cell (PBMC) DNA at treatment failure while on therapy and after stopping therapy. DESIGN: Fifty-eight patients failing their first highly active antiretroviral treatment while on therapy and 50 patients after a median of 18.6 weeks after treatment interruption following multiple treatment failures were analysed. METHODS: Paired plasma HIV-1 RNA and PBMC HIV-1 DNA were used for genotypic antiretroviral resistance testing. Drug resistance was computed by using the Stanford on-line genotype interpretation system. RESULTS: The extent of drug resistance was larger in plasma RNA than in PBMC DNA in the on-therapy group (P=0.004) and in PBMC DNA than in plasma RNA in the off-therapy group (P=0.04). Interpretation of genotype based on PBMC DNA and plasma RNA in the on-therapy and in the off-therapy group would have missed detection of resistance to one or more drugs in 21 and 22% of the patients, respectively, compared to interpretation based on the other blood compartment. In a subset of 27 patients for whom a sample before stopping therapy was available, there was a significant decrease in the number of RNA (mean 8.1 to 5.3, P=0.004), but not DNA (mean 6.8 to 5.7, P=0.143), resistance mutations following treatment interruption. CONCLUSION: While plasma RNA is the material of choice for early detection of drug resistance while on therapy, analysis of PBMC DNA may additionally support and possibly improve sensitivity of resistance testing in the absence of therapy. PMID- 12553479 TI - Transient rebounds of low-level viraemia among HIV-infected patients under HAART are not associated with virological or immunological failure. AB - OBJECTIVE: To analyse whether blips are associated with a higher risk of virological or immunological failure than persistent undetectable viraemia (UND) among HIV-infected patients receiving HAART. DESIGN: Retrospective cohort study. SUBJECTS: Patients with blips or UND were selected from a prospective cohort of 330 patients under HAART for over 48 weeks. Blips were defined as detectable viraemia up to a maximum of 1000 copies/ml preceded by two consecutive visits and followed by one visit showing undetectable viraemia. Patients were included according to the following criteria: i) Blip group: patients that showed transient relapses of viraemia; ii) UND group: patients who had achieved UND on HAART before 24 weeks of therapy and that sustained viral suppression for four consecutive visits. MAIN OUTCOME MEASURES: Virological and immunological failure. RESULTS: Thirty seven (11%) and 65 (20%) patients showed blips and persistent UND, respectively. Virological failure was observed in three (8.1%) patients in the blip group and 11 (16.9%) patients in the UND group (P=0.25). The time to virological failure was shorter in the UND group (P=0.12). The rates of virological failure and the time to virological failure were similar between both groups after excluding patients with compliance <95%. The time to immunological failure was also similar in both groups (P=0.5). In a Cox model, only the use of saquinavir hard gel-based regimens was independently associated with the time to virological and immunological failure. CONCLUSION: Patients under HAART with transient low-level viraemia are not at an increased risk of developing virological or immunological failure. PMID- 12553480 TI - Lifetime cost of HIV care in France during the era of highly active antiretroviral therapy. AB - OBJECTIVE: To estimate the treatment and health care costs of HIV infection or AIDS in France during the era of highly active antiretroviral therapy (HAART). DESIGN: We used a clinical database of HIV-infected patients to calculate the resource use and cost of care for different stages of HIV infection. Costs were incorporated into a computer-based, probabilistic simulation model of the natural history and treatment of HIV infection to estimate the lifetime cost of treating patients with HIV disease. SETTING: A northern France HIV clinical cohort. PARTICIPANTS: 1232 HIV-infected patients followed from January 1994 through July 1998. RESULTS: In the absence of an AIDS-defining event, the average total cost of care ranged from 670 euros (1 euro=US $1.19) per person-month in the highest CD4 stratum (>500/microl) to 1060 euros per person-month in the lowest CD4 stratum (< or = 50/microl). The mean cost of care was estimated at 3370 euros per person-month during the initial months around the occurrence of an AIDS-defining event; at 1750 euros per person-month during the period spanning from 2 months after the diagnosis of specific AIDS-defining event to 1 month prior to death; and at 13010 euros per person-month in the final month prior to death. If clinical management of HIV infection began at a CD4 cell count of 378/microl, as in this cohort, the discounted lifetime cost of treating an HIV-infected French patient was estimated at 214000 euros. The undiscounted costs were 309000 euros over a projected life expectancy of 16.4 years. CONCLUSION: The cost of HIV disease varies widely depending upon the stage of illness. These estimates of stage-specific and lifetime costs of HIV care will assist health policy planners in assessing the burden of disease in the era of HAART and projecting future resource requirements. PMID- 12553481 TI - Single-dose pharmacokinetics of enteric-coated didanosine in HIV-infected children. AB - Didanosine remains a cornerstone nucleoside analogue for the treatment of HIV infection. A potential problem with the buffered formulations of didanosine is the likelihood of interactions with other drugs that require an acidic pH for absorption or can be chelated by cations in the buffer. An encapsulated enteric coated (EC) bead formulation of didanosine has been approved and is routinely used as an alternative to the chewable/dispersible buffered tablet formulation. The objective of this study was to evaluate the single-dose pharmacokinetics of didanosine EC at 240 mg/m2 in 10 HIV-infected children. Didanosine EC was administered at time 0 on an empty stomach with no other concomitant medications. Blood samples were collected at pre-dose, 0.5, 1, 2, 4, 8 and 12 h post-dose. Didanosine was measured in plasma using radioimmunoassay. Ten subjects completed the intensive pharmacokinetic evaluation; data are available for eight participants. Plasma concentrations of didanosine following EC administration were analysed using non-compartmental methods. Median (range) AUCinfinity, Cmax, Tmax and CL/F for didanosine following EC administration were 2385 (1291, 3966) ng x h/ml, 854 (300, 1799) ng/ml, 3.0 (1.0, 8.1) h and 3.3 (2.7-6.4) l/h/kg, respectively. Results from this study indicate that the didanosine Cmax is decreased and Tmax is prolonged, but total exposure of didanosine in plasma following didanosine EC administration appears similar to previous data collected in HIV-infected children following buffered didanosine administration. PMID- 12553482 TI - Analysis of virological efficacy in trials of antiretroviral regimens: drawbacks of not including viral load measurements after premature discontinuation of therapy. AB - OBJECTIVES: To compare two analytic approaches to assess the virological effect of HAART according to the intention-to-treat (ITT) principle. MATERIAL: Data from 2318 patients enrolled in 10 randomised clinical trials (RCTs) and from 3091 patients followed in an observation cohort (EuroSIDA) starting their first HAART regimen. METHODS: Two classifications of defining virological response 48 weeks after starting the therapy to be evaluated were compared: 1) only patients remaining on the therapy and having a plasma viral load (pVL) below a given cut off level at week 48 were classified as responders (ITT/s=f); and 2) patients with a pVL below a given cut-off at week 48 whether they remained on initial assigned therapy or switched therapy were responders (ITT/s incl). In both analyses, patients with missing data at week 48 were classified as failures (i.e., non-responders). RESULTS: According to ITT/s=f, 22-70% of the patients starting a HAART regimen in a RCT experienced a virological response at week 48. Only two RCTs had complete follow-up data (n=424): between 29 and 62% achieved a virological response at week 48 in the six treatment arms evaluated in the studies according to ITT/s=f, and 41-72% according to ITT/s incl. Among those who discontinued the therapy to be evaluated in these two trials, 13-45% (cohort: 39 74%) subsequently experienced a virological response at week 48. The subsequent response rates were associated with the reason for discontinuation (toxicity versus confirmed virological failure: 63 vs 33%), varied largely across regimens and were not associated with the discontinuation rate. CONCLUSIONS: Discontinuation of follow-up at switch from the therapy to be evaluated remains common in antiretroviral treatment trials, but leads to an imprecise and incomplete assessment of the intrinsic effect of a given regimen. Complete follow up of all patients should be encouraged strongly as this will allow for several complementary analytic approaches and a focus on optimal treatment strategies rather than specific regimens. PMID- 12553483 TI - Genotypic and phenotypic resistance patterns in early-stage HIV-1-infected patients failing initial therapy with stavudine, didanosine and nevirapine. AB - The objectives of this study were to determine the genotypic and phenotypic patterns of resistance in a group of early-stage antiretroviral-naive patients failing initial therapy with didanosine, stavudine and nevirapine. These patterns of resistance were determined at baseline and at time of virological failure in 89 antiretroviral-naive patients with CD4 cells >500 cells/ml and viral load >5000 copies/ml who received initial antiretroviral therapy with didanosine plus stavudine and nevirapine as part of the SCAN study, and who failed after having reached undetectable plasma levels (<200 copies/ml). Of the 89 patients recruited in the SCAN study, 14 (16%) developed a virological failure after reaching a viral load below 200 copies/ml after a median of 20 months of follow-up. At baseline, none of these 14 patients had genotypic resistance. At time of failure, six out of 14 (43%) failing patients had wild-type genotype and no phenotypic resistance. Suboptimal compliance could be documented in four of these six patients. Seven patients (50%) had nevirapine resistance mutations (mainly K103N [4/7], Y181C/I [2/7], G190A/S [2/7] and V108I [1/7]) associated with phenotypic high-level resistance to nevirapine, delavirdine and efavirenz (nevirapine >47.4- to 58.1-fold, delavirdine >74.4- to 168.9-fold and efavirenz >56.0- to 347.2 fold). Four of these seven patients also had thymidine analogue-associated mutations (TAM) (T215Y/F [2/4], M41L [1/4], D67N [2/4] and K70R [1/4]). Finally, one patient (7%) had exclusively TAM mutations (M41L). None of the patients developed mutations associated with didanosine resistance or phenotypic resistance to didanosine or stavudine. Suboptimal compliance or selection of nevirapine resistance often with TAM mutations was frequently associated with virological failure in a cohort of early-stage chronic HIV-1-infected patients treated with a protease inhibitor-sparing regimen. PMID- 12553484 TI - No protective effect of acyclovir on HIV disease progression in a cohort of HSV-2 HIV-infected individuals. AB - The efficacy of anti-herpetic drugs in decreasing HIV disease progression has not been clarified. We studied a cohort of 126 HIV-positive individuals with known date of seroconversion who were HSV-2-seropositive to determine if progression to AIDS was influenced by treatment with acyclovir. In the multivariate analysis, being homosexual and low CD4 count were associated with a faster progression to AIDS, whereas treatment with acyclovir showed a 37.0% protective effect compared to those who did not receive it when antiretroviral treatment was not included in the model. When including antiretroviral therapy, the protective effect of acyclovir decreased to 9.0% and that of antiretroviral therapy was 43.0% for monotherapy and 36.0% for double therapy, suggesting that most of the protective effect of acyclovir in the previous model was due to antiretroviral therapy. In conclusion, treatment with acyclovir does not seem to prolong significantly survival to AIDS among HIV-positive individuals who are HSV-2-infected. PMID- 12553485 TI - Evolution of antiretroviral phenotypic and genotypic drug resistance in antiretroviral-naive HIV-1-infected children treated with abacavir/lamivudine, zidovudine/lamivudine or abacavir/zidovudine, with or without nelfinavir (the PENTA 5 trial). AB - PURPOSE AND METHODS: To describe the evolution of resistance to zidovudine (ZDV), lamivudine (3TC), abacavir (ABC) and nelfinavir (NFV), 113 previously untreated children in the PENTA 5 trial had resistance assayed at baseline, rebound and/or 24, 48, 72 weeks (VIRCO: phenotyping and genotyping with 'Virtual Phenotype' interpretation). RESULTS: At baseline, few reverse transcriptase mutations and no primary protease inhibitor mutations were observed. Time to detectable HIV-1 RNA with reduced phenotypic susceptibility to any drug was shortest in the ZDV+3TC arm (overall logrank P=0.02). Through a median follow-up of 55 weeks, at their last assessment 11 (28%), 16 (40%) and 13 (32%) children with detectable HIV-1 RNA and a resistance test available had mutations conferring resistance to none, one, or two or more trial drugs, respectively, according to the virtual phenotype. Reduced phenotypic susceptibility to ABC only occurred in the 3TC+ABC arm and required K65R and/or L74V in addition to M184V. NFV-resistant virus was selected slowly through D30N or L90M pathways, and selection of ZDV-resistant virus was rare. CONCLUSIONS: Selection of 3TC-resistant virus was most frequent, followed by NFV and/or ABC; selection of ZDV-resistant virus was rare. Importantly, although in vitro, ABC selects for M184V as the first mutation, ABC did not select for M184V when combined with ZDV without 3TC. The most sustained HIV-1 RNA response was in the 3TC+ABC arm, but mutations conferring reduced susceptibility to 3TC and/or ABC evolved more frequently if virological failure occurred with 3TC+ABC than with ZDV+ABC. PMID- 12553486 TI - Long-term virological outcome in patients infected with multi-nucleoside analogue resistant HIV-1. AB - The emergence of HIV strains that are resistant to antiretroviral drugs is a major cause of treatment failure. Two sets of mutations: the Q151 M complex and the 69 insert, cause resistance to multiple nucleoside analogues. We report the response to treatment in 12 patients with multiple NRTI-resistant HIV-1 strains. Seven of 12 patients (58%) were able to maintain a viral load below 200 copies/ml at week 48. The patients most likely to obtain therapeutic success were those having no or low-level resistance to non-nucleoside reverse transcriptase inhibitors and/or protease inhibitors. New and more effective drugs are needed for patients with HIV-1 that is resistant to more than one of the current three classes of HIV drugs. PMID- 12553487 TI - Venous and arterial blood lactate in HIV-infected patients. PMID- 12553488 TI - Microglia in diseases of the central nervous system. AB - Microglia (MG) are enigmatic cells of the central nervous system (CNS). MG are morphologically, antigenically and functionally flexible, and have the potential for mobility and proliferation. MG are professional antigen-presenting cells and constitute part of the local CNS innate immune system, communicating with other immune cells via chemokines, cytokines and growth factors. MG contain several antigenic and functional markers similar to macrophages and dendritic cells (DCs), but also present several differences from DCs. The exact role(s) played by MG in the normal human CNS is the topic of lively debate. MG participate in many reactive processes in the CNS and are therefore an integral part of lesions in a variety of pathologic conditions. It is thought that MG may exacerbate diverse neurological conditions, including viral encephalitis, AIDS, Multiple Sclerosis (MS) and Alzheimer's disease. A recurrent theme is the perpetuation by MG of pathological cycles of monocyte recruitment, activation and cytopathic secretions, and/or auto antigen presentation. PMID- 12553489 TI - Progress in producing knockout models for xenotransplantation by nuclear transfer. AB - The pig is thought to be the most suitable non-human organ source for xenotransplantation. However, one of the major constraints to using pig organs for xenotransplantation is human natural antibody-mediated hyperacute rejection (HAR). Elimination of alpha(1, 3) galactosyltransferase (GGTA1) from the pig is expected to be a solution to the problem of hyperacute and delayed vascular rejection. Based on the extensive efforts made in characterization of GGTA1 in structure and function, improvement in the technique of DNA transfection of somatic cells and optimization of the pig nuclear transfer (NT) procedures, a specific modification has been made to one copy of the GGTA1 gene thus rendering it non-functional. When homozygosity of the genetic modification is achieved, this will allow concentrated efforts to address the role of galalpha(1, 3) gal specific natural antibody in HAR, acute vascular rejection and to examine the efficacy of xenotransplantation in a non-human primate model without HAR confounding the experiments. Since swine are widely used as models of human disease, the ability to make specific genetic modifications to swine as described here is predicted to have a large impact not only on production agriculture but also on biomedicine. PMID- 12553490 TI - Angiotensin receptor antagonists in patients with nephropathy due to type 2 diabetes. AB - In the past few years diabetes has become the leading cause of end-stage renal disease in all Western countries. A correlation between blood pressure and rate of progression in diabetic nephropathy was noted very early, and increased local activity of the renin angiotensin system was identified as a major pathophysiological mechanism for proteinuria and nephrosclerosis in diabetic patients. Angiotensin converting enzyme (ACE) inhibitors have been shown to slow progression of nephropathy in type 1 diabetic patients. The majority of diabetic patients with nephropathy, however, are suffering from type 2 diabetes and until last year there was no convincing evidence of ACE inhibitors being able to slow progression in type 2 diabetic patients with nephropathy. Three new studies now fill this gap, showing that angiotensin receptor blockers (ARB) are nephroprotective in patients with type 2 diabetes, independently of blood pressure. This review provides an in-depth discussion of the results of these studies and provides recommendations for patient management. PMID- 12553491 TI - Optimal treatment and current situation in reperfusion after thrombolysis for acute myocardial infarction. AB - Acute myocardial infarction is the leading cause of death in the industrialized world and the paramount goal is establishing early, complete, and sustained reperfusion at the myocardial tissue level. For hospitals without the capacity to perform emergent percutaneous coronary intervention, fibrinolytic therapy plays a critical role although it is limited by a 67% success rate. Despite promising pilot studies, reduced-dose fibrinolytic therapy with glycoprotein IIb/IIIa therapy (GUSTO-V) and full-dose fibrinolytic therapy with enoxaparin (ASSENT-3) or bivalirudin (HERO-2) provide only marginally improved clinical outcomes. Adjunctive in-hospital and secondary preventive measures should include an aspirin, a beta-blocker, an ACE inhibitor, and a statin, based on the Heart Protection Study, unless contraindicated. Patients should be risk stratified, participate in a cardiac rehabilitation program, cease smoking tobacco, and have an intracardiac defibrillator (ICD) implanted if their LV systolic function is < or = 30% at one month based on the MADIT-2 trial. PMID- 12553492 TI - Mapping genetic influences on human brain structure. AB - Recent advances in brain imaging and genetics have empowered the mapping of genetic and environmental influences on the human brain. These techniques shed light on the 'nature/nurture' debate, revealing how genes determine individual differences in intelligence quotient (IQ) or risk for disease. They visualize which aspects of brain structure and function are heritable, and to what degree, linking these features with behavioral or cognitive traits or disease phenotypes. In genetically transmitted disorders such as schizophrenia, patterns of brain structure can be associated with increased disease liability, and sites can be mapped where non-genetic triggers may initiate disease. We recently developed a large-scale computational brain atlas, including data components from the Finnish Twin registry, to store information on individual variations in brain structure and their heritability. Algorithms from random field theory, anatomical modeling, and population genetics were combined to detect a genetic continuum in which brain structure is heavily genetically determined in some areas but not others. These algorithmic advances motivate studies of disease in which the normative atlas acts as a quantitative reference for the heritability of structural differences and deficits in patient populations. The resulting genetic brain maps isolate biological markers for inherited traits and disease susceptibility, which may serve as targets for genetic linkage and association studies. Computational methods from brain imaging and genetics can be fruitfully merged, to shed light on the inheritance of personality differences and behavioral traits, and the genetic transmission of diseases that affect the human brain. PMID- 12553493 TI - Stereochemistry and drug efficacy and development: relevance of chirality to antidepressant and antipsychotic drugs. AB - Many drugs contain a chiral center or a center of unsaturation, or such centers result during metabolism of these drugs. Often such drugs are marketed as a mixture of the resultant enantiomers (racemates) or of geometric isomers, respectively. These enantiomers (molecules that are not superimposible on their mirror image) or geometric isomers may differ markedly from each other with regard to their pharmacodynamic and/or pharmacokinetic properties. This review deals primarily with drugs with chiral centers, and possible complications arising from the use of racemates are discussed. Recent developments in resolution of enantiomers, increased knowledge of the molecular structure of specific drug targets and a heightened awareness of several possible advantages of using single enantiomers rather than racemic mixtures of drugs have led to an increased emphasis on understanding the role of chirality in drug development. This has resulted in increased investigation of individual enantiomers early on in the development of drugs and in 'chiral switching', i.e. the replacement of a racemate of a drug which has already been approved or marketed by a single enantiomer. Although stereochemistry is an important matter to consider in drugs of virtually all classes, this review focuses on the relevance of chirality to antidepressant and antipsychotic drugs. Examples of the effects of chiral centers on the properties of antidepressants (tricyclics, selective serotonin reuptake inhibitors, monoamine oxidase inhibitors, viloxazine, bupropion, mianserin, venlafaxine, mirtazapine and reboxetine), antipsychotics and/or some of their metabolites are discussed. PMID- 12553494 TI - Quantitation of erbB2 positivity for evaluation of high-risk patients. AB - BACKGROUND: Because trastuzumab therapy is expected to be effective in a large fraction of erbB2 (HER-2/neu) overexpressing breast cancers, it is important to find the optimal method for evaluation of erbB2 positivity, and the patient group at greatest risk of dying without this therapy. AIM: We evaluated erbB2 immunopositivity in breast cancer with the aim of finding a high-risk group for primary trastuzumab therapy. METHODS: Three hundred and seventeen samples were evaluated with an immunostaining index. Optimal cut point was systematically tested, and the effect of bcl-2 status on survival in the high-risk group was studied. RESULTS: Among N+ patients the index value 1.5 reflected the biggest difference in survival. There was a significant correlation between erbB2 positivity and bcl-2 negativity. ErbB2 was a prognosticator among postmenopausal, N+, and postmenopausal N+ patients. In multivariate analysis, erbB2 was the best prognosticator among postmenopausal N+ patients. Six out of seven N+ patients with erbB2 index 1.5 or above died including all postmenopausal patients. Bcl-2 positivity was associated with longer survival in the erbB2 positive patient group. CONCLUSIONS: The most obvious patients for primary trastuzumab therapy in breast cancer are N+ patients with high erbB2 immunostaining index (> 1.5) and bcl-2 negative immunostaining. In our material 2% of all breast cancer patients fell in this category. This patient group should be selected for testing trastuzumab in the primary treatment. PMID- 12553495 TI - Effects of thyroid hormone suppression therapy on adverse clinical outcomes in thyroid cancer. AB - BACKGROUND: Long-term thyroid hormone (TH) therapy aiming at the suppression of serum thyrotropin (TSH) has been traditionally used in the management of well differentiated thyroid cancer (ThyrCa). However, formal validation of the effects of thyroid hormone suppression therapy (THST) through randomized controlled trials is lacking. Additionally, the role - if any - of TSH effect at low ambient concentrations upon human thyroid tumorigenesis remains unclear. AIM: Evaluation of the effect of THST on the clinical outcomes of papillary and/or follicular ThyrCa. METHODS: By using a quantitative research synthesis approach in a cumulative ThyrCa cohort, we evaluated the effect of THST on the likelihood of major adverse clinical events (disease progression/recurrence and death). A total of 28 clinical trials published during the period 1934-2001 were identified; only 10 were amenable to meta-analysis. Causality was assessed by Hill criteria. RESULTS: Out of 4, 174 patients with ThyrCa, 2, 880 (69%) were reported as being on THST. Meta-analysis showed that the group of patients who received THST had a decreased risk of major adverse clinical events (RR = 0.73; Cl = 0.60-0.88; P < 0.05). Further, by applying a Likert scale, 15/17 interpretable studies showed either a 'likely' or 'questionable' beneficial effect of THST. Assessment of causality between TSHT and reduction of major adverse clinical events suggested a probable association. CONCLUSIONS: THST appears justified in ThyrCa patients following initial therapy. As most primary studies were imperfect, future research will better define the effect of THST upon ThyrCa clinical outcomes. PMID- 12553496 TI - Increased DNA adducts in Barrett's esophagus and reflux-related esophageal malignancies. AB - BACKGROUND: DNA adduct formation can initiate carcinogenic processes. AIM: To examine the pre-malignant condition of Barrett's esophagus by measuring the DNA adducts. METHODS: DNA adducts were measured in the proximal and distal esophagus of patients with Barrett's esophagus (n = 9), patients with adenocarcinoma in the distal esophagus/esophagogastric junction (n = 28), and in control group of patients (n = 8) using the 32-P-postlabeling method. The average levels of DNA adducts are expressed as mean adducts/10(9) nucleotides + standard error of the mean. RESULTS. The average DNA adduct levels in the distal esophagus were significantly higher in both the Barrett's esophagus (24.5 +/- 7.9) and the adenocarcinoma (12.0 + 3.0) than in the control patients (0.1 +/- 0.08), P < 0.001. In the proximal esophagus, the DNA adduct levels were approximately equal in the Barrett's esophagus (7.0 +/- 1.0) and in the adenocarcinoma group (6.4 +/- 0.65). However, the levels in the proximal esophagus in both groups were significantly higher than in the control group (2.1 +/- 0.67), P < 0.05. CONCLUSIONS: Patients with Barrett's esophagus and patients with esophageal/esophagogastric junction adenocarcinoma had significantly more DNA adducts than the control group. These results support the current concept of the carcinogenic potential of chronic gastroesophageal reflux, and the pre-malignant condition of Barrett's esophagus. PMID- 12553497 TI - Animal and human models for sepsis. AB - Several preclinical models for sepsis have been used in the last decades to successfully unravel the pathophysiologic processes during sepsis. Furthermore, these models for sepsis revealed promising immunomodulating agents for the treatment of sepsis. Nevertheless, several clinical trials evaluating the efficacy of these new anti-inflammatory agents in septic patients showed disappointing results. In this article the advantages and disadvantages of different models for sepsis are discussed. Most models for sepsis lack an infectious focus. Importantly, investigations studying the effects of several immunomodulating strategies have demonstrated strikingly opposite results when using models for sepsis lacking an infectious focus and when using models for sepsis with a more natural route of infection. These differences will be discussed in this article. In general, it is advised to use a combination of models to test a new therapeutic agent, before starting a clinical study evaluating this new therapy. PMID- 12553498 TI - The use of corticosteroids in severe sepsis and acute respiratory distress syndrome. AB - During sepsis or acute respiratory distress syndrome, the hypothalamic pituitary adrenal axis is rapidly activated through a systemic pathway, i.e. by circulating pro-inflammatory cytokines and through the vagus nerve. Subsequently, the adrenal glands release cortisol, a hormone which will likely counteract the inflammatory process and restore cardiovascular homeostasis. Both experimental models and studies in humans suggest that inadequate hypothalamic pituitary adrenal axis response to stress accounts, at least partly, for the genesis of shock and organ dysfunction in sepsis and acute respiratory distress syndrome. Relative adrenal insufficiency and peripheral glucocorticoid resistance syndrome are the two main features of the inappropriate hormonal response and provide the grounds for cortisol replacement in these diseases. In practice, a high dose of corticosteroids (i.e. one to four boluses of 30 mg/kg of methylprednisolone, or equivalent) had no effects on survival in severe sepsis or acute respiratory distress syndrome. There are at least seven randomised controlled trials reporting the benefits and risks of low dose corticosteroids (i.e. 200 to 300 mg daily of hydrocortisone or equivalent) given for a prolonged period in severe sepsis or in the late phase of acute respiratory distress syndrome. These trials showed consistently that, in these patients, the use of low dose of corticosteroids alleviated inflammation, restored cardiovascular homeostasis, reduced organ dysfunction, improved survival and was safe. Further studies are ongoing to better identify the target population. In the meantime, cortisol replacement (i.e. 200 to 300 mg daily of hydrocortisone or equivalent) should be considered as standard care for these patients. PMID- 12553499 TI - Treatment of impaired perfusion in septic shock. AB - Severe sepsis and septic shock are relatively common problems in intensive care. The mortality in septic shock is still high, and the main causes of death are multiple organ failure and refractory hypotension. Impaired tissue perfusion due to hypovolemia, disturbed vasoregulation and myocardial dysfunction contribute to the multiple organ dysfunction. Treatment of hemodynamics in septic shock consists of appropriate fluid therapy guided by invasive monitoring combined with vasoactive drugs aiming to correct hypotension and inappropriately low cardiac output. The drug of choice for low vascular resistance is norepinephrine, while insufficient myocardial contractility is commonly treated with dobutamine. The use of norepinephrine seems to be associated with better prognosis as compared to results from the use of dopamine or epinephrine. In septic shock, vasopressin levels are low, and therefore, vasopressin has been advocated as a vasopressor. Its effectiveness and safety have not yet been documented, and so far it is regarded as an experimental treatment Recent data support the use of corticosteroid, at least in some of the patients with septic shock. Also, activated protein C, a drug with anti-inflammatory and antithrombotic properties, decreases mortality in patients with septic shock. PMID- 12553500 TI - Protein C pathway in sepsis. AB - The goals of this chapter are to provide a brief review of the biology of the protein C pathway and some of the features of the pathway that make it uniquely positioned to control microvascular coagulation and control the acute inflammatory response. Activated protein C works as an antithrombotic agent by inactivating factors Va and VIIIa. It is particularly effective at preventing microvascular thrombosis. Platelets may provide a margin of safety for activated protein C as an antithrombotic. Approximately 25% of the factor V/Va in plasma is contained within the platelet and hence resistant to time dependent inactivation by activated protein C. In addition, factor Va bound to the platelet surface is relatively resistant to inactivation by activated protein C. Activated protein C also facilitates clot lysis by inhibiting plasminogen activator inhibitor 1, a process that is accelerated markedly by vitronectin. Inflammatory cytokines like tumor necrosis factor alpha (TNFalpha) and interleukin-1beta (IL-1beta) downregulate two key components of the protein C activation complex, thrombomodulin and the endothelial cell protein C receptor resulting in decreased protein C activation. Activated protein C in turn has been shown in several animal models and in vitro to inhibit TNF elaboration in response to endotoxin. This inhibition appears to be due to diminished nuclear factor kappaB (NF kappaB) expression and nuclear translocation. Activated protein C has been shown to reduce the rate of death due to severe sepsis. This reduction may be due to both the anticoagulant effects as demonstrated by a reduction in D-dimer and inflammatory effects as demonstrated by a reduction in interleukin 6. PMID- 12553501 TI - Management of septic shock. AB - Severe sepsis is a common disease process affecting some 2-11% of hospital admissions in the US. Severe sepsis and septic shock are associated with considerable morbidity and mortality, and account for a large part of intensive care unit costs. Until recently, the management of septic shock relied on the treatment of underlying infection with antimicrobial agents and surgical removal of any infectious source, and individual support of failing organs. However, in the last few years we have seen huge strides being made in our understanding of the pathophysiology of the sepsis response, and in our ability to manipulate that response. In the last couple of years these advances have come to fruition with the development of a drug, drotrecogin alfa, which specifically reduces mortality from this all too often fatal disease. While intensive early resuscitation remains the cornerstone of management, new approaches are beginning to form part of sepsis management protocols and will lead to improved outcomes for patients with this disease process. PMID- 12553502 TI - Markers of inflammation in sepsis. AB - Pathophysiology of sepsis is characterised by a whole body inflammatory reaction and concurrent activation of the host's anti-inflammatory mechanisms. The balance between pro- and anti-inflammatory reactions is critical for the outcome of the patient. Strongly activated phagocytes and high levels of proinflammatory cytokines occur in patients who are at risk of developing circulatory shock and multiple organ dysfunction. Extensive anti-inflammatory reaction, which is characterised by the presence of high levels of circulating anti-inflammatory cytokines and impaired innate and adaptive immune functions, renders critically ill patients prone to secondary infections. Evaluation of the immune-inflammatory status on admission to the hospital may be helpful in the early identification of patients who are bound to develop organ dysfunction. Such patients could possibly benefit from a mode of therapy aimed at modifying the course of inflammatory response. The use of inflammatory markers may also improve diagnosis of severe infection. The present review summarises the studies on markers of inflammation and immune suppression used, first, as predictors of organ dysfunction in patients with systemic inflammation, and, second, as indicators of infection in adults and neonates. PMID- 12553503 TI - Meningococcal disease: treatment and prevention. AB - Many countries have been experiencing a significant increase in meningococcal disease. With the strains currently circulating, septicaemia is now a more frequent manifestation than meningitis and early recognition of disease manifestations by patient, parent or physician as well as early recognition of disease severity are the most important factors in attempting to reduce mortality and morbidity. Ceftriaxone is the treatment of choice but must be accompanied by aggressive supportive therapy in those with severe disease. The role of steroids is unknown. The evidence to support their use in both meningitis and severe systemic sepsis is discussed. The purified polysaccharide vaccines that have been available for some years may play a limited role in disease prevention. The recently introduced conjugate vaccine for preventing serogroup C disease represents a major advance but no vaccine is currently available to prevent serogroup B disease, cases of which will continue to challenge clinical practice. PMID- 12553504 TI - Primary hemiarthroplasty in old patients with displaced femoral neck fracture: a 1-year follow-up of 103 patients aged 80 years or more. AB - 103 patients with displaced femoral neck fractures (Garden 3-4) treated with primary hemiarthroplasty in the Department of Orthopedics, Malmo University Hospital, Sweden 1998-1999 were followed in a prospective, consecutive study for 1 year. Inclusion criteria were age of at least 80 years, signs of mental changes and/or residence in an institution. The control group consisted of 69 patients with internal fixation (Hansson hook pins). The arthroplasty group required more blood transfusions, a longer operation and had more superficial infections. No differences were detected as regards other complications, length of hospital stay, in-hospital mortality or ability to return home. The 1-year mortality rates were similar in the arthroplasty (29/103) and control groups (28/69). Within 1 year, we found a lower failure rate in the arthroplasty group (6/103) than in the control group (18/69). In the arthroplasty group, 2/103 had dislocations. Of the surviving arthroplasty patients at 12 months, 31/74 could walk as well as before the fracture and 55/74 had no pain. We recommend primary hemiarthroplasty for demented and/or institutionalized patients over 80 years with displaced femoral neck fractures. PMID- 12553505 TI - Prophylaxis of heterotopic ossification after total hip arthroplasty: a prospective randomized study comparing indomethacin and meloxicam. AB - We performed a randomized, prospective study on the prophylaxis of heterotopic ossification (HO) after total hip arthroplasty (THR), comparing indomethacin and the selective COX-2 inhibitor meloxicam. From the day after surgery, 272 patients were treated with 7.5 mg meloxicam, 15 mg meloxicam, or 2 x 50 mg indomethacin a day, for 14 days. After 6 months, radiographs of patients treated with 7.5 mg meloxicam showed that HO had occurred in one third. This treatment was therefore stopped after 26 patients have been assigned to this group. According to the intention-to-treat principle, patients given 15 mg meloxicam developed HO in 25% (20% Brooker grade I, 4% grade II and 1% grade III) and those given indomethacin in 10% (7% Brooker grade I, 1% grade II and 2% grade III), a statistically significant difference. PMID- 12553506 TI - Acute total hip arthroplasty for acetabular fractures in the elderly: 11 patients followed for 2 years. AB - We describe the use of cable fixation and acute total hip replacement for acetabular fracture in the elderly. 12 patients with acetabular fractures, having a mean age of 79 (65-93) years, were treated with cable fixation and acute total hip arthroplasty. 8 were T-shaped fractures and 4 associated fractures of the posterior column and posterior wall. 1 patient died 5 months after surgery and the remaining 11 were followed for 2 years. All patients had a good clinical outcome. Radiographic assessment showed healing of the fracture and a satisfactory alignment of the cup without loosening. This surgical technique provides good primary fixation, stabilizes complex acetabular fractures in elderly patients with osteoporotic bone and permits early postoperative mobilization. PMID- 12553507 TI - Cable fixation in displaced fractures of the acetabulum: 21 patients followed for 2-8 years. AB - We evaluated the outcome in 21 patients having acetabular fractures who had been treated with a 2 mm-braided cable, alone (n = 6), or together with plate and screws. Reduction with a fracture gap of less than 2 mm was achieved in 20 cases. The operation lasted an average of 3 (1.5-4.8) hours. The average blood loss was 1.4 (1.3-2.8) L. No intraoperative complication occurred in relation to surgery. Postoperative complications included 2 cases each of posttraumatic arthritis and avascular necrosis of the femoral head. After a mean follow-up of 4 (2-8) years, the clinical outcome was excellent in 13, good in 3, fair in 3 and poor in 2, and the radiographic outcome was excellent in 10, good in 6, fair in 1 and poor in 4. We believe that cable fixation is a useful technique since the indirect reduction requires a limited exposure, avoids penetration of the screw into the joint, and improves fixation in patients with osteoporosis. PMID- 12553508 TI - No effect of drains on the postoperative hematoma volume in hip replacement surgery: a randomized study using scintigraphy. AB - In a prospective randomized study we used erythrocyte scintigraphy to evaluate whether drainage reduced the hematoma volume after total hip arthroplasty. 12 patients were left without drains and 10 patients had two drains inserted, one below the fascia and one subcutaneously. We used tomographic registration (SPECT) to calculate the volume of the hematoma (erythrocytes) about 22 hours after surgery and found that drainage did not reduce the volume, but increased the need for blood transfusion. PMID- 12553509 TI - Stemmed femoral knee prostheses: effects of prosthetic design and fixation on bone loss. AB - Although the revision rates for modern knee prostheses have decreased drastically, the total number of revisions a year is increasing because many more primary knee replacements are being done. At the time of revision, bone loss is common, which compromises prosthetic stability. To improve stability, intramedullary stems are often used. The aim of this study was to estimate the effects of a stem, its diameter and the interface bonding conditions on patterns of the bone remodeling in the distal femur. We created finite element models of the distal half of a femur in which 4 types of knee prostheses were placed. The bone remodeling process was simulated using a strain-adaptive bone remodeling theory. The amount of such remodeling was determined by calculating the changes in bone mineral density in 9 regions of interest from simulated DEXA scans. The computer simulation model showed that revision prostheses tend to cause more bone resorption than primary ones, especially in the most distal regions. Predicted long-term bone loss due to a revision prosthesis with a thin stem equalled that around a prosthesis with an intercondylar box. However, strong regional differences were found--the stemmed prostheses having more bone loss in the most distal areas and some bone gain in the more proximal ones. A prosthesis with a thick stem led to an increase in bone loss. When the prosthesis-cement interface was bonded, more bone loss was predicted than with an unbonded interface. These results suggest that a stem which increases stability initially may reduce stability in the long term. This is due to an increase in stress shielding and bone resorption. PMID- 12553510 TI - Tibiofemoral contact stress after total knee arthroplasty: comparison of fixed and mobile-bearing inlay designs. AB - We measured tibiofemoral contact stresses and the load-bearing contact area of fixed and mobile-bearing inlay knee prostheses under dynamic loading conditions. An electronic resistive pressure-measuring sensor was used to detect contact stresses and contact area in five cadaver knees. Stresses were measured with the tibial component aligned normally, as well as in internally- and externally rotated positions. The average peak contact stresses measured on the fixed inlay were greater (medial 21 MPa and lateral 21 MPa) than those on the mobile inlay (medial/lateral 7.7/5.3 MPa, p = 0.04). Although the average peak contact stresses of the fixed standard inlay greatly exceeded the contact stresses of the other two inlay designs in each malrotated position tested, no statistically significant differences were seen. The data suggest that the ability of the inlay to translate on the tibial baseplate permits the inlay to align itself on the femoral component so that the contact surface area is maximized and contact stresses are reduced. PMID- 12553511 TI - Kinetic and kinematic characteristics of gait in patients with medial knee arthrosis. AB - We compared the mechanics of gait in 13 patients with early medial arthrosis (OA) of the knee and 13 normal controls, by measuring gait events, kinematic and kinetic parameters. In the OA group, walking velocity, cadence and stride length were reduced and stride time and double support time accordingly increased on both sides, the overall stance phase was prolonged in the OA group, but the stance phase and swing phase peak flexion were reduced. The varus in the stance phase and the valgus in the swing phase were increased. The extensor moment in the loading response was increased and the flexor moment at late stance reduced in the OA group. Patients with OA had a greater valgus (abductor) and internal rotation moment during the stance phase. The times to second vertical force peak (VFP) were similar in the two groups. Values of VFP1 and VFP2 were lower in the OA group. Our findings indicate that computerized gait analysis can be used to reveal various mechanical abnormalities accompanying arthrosis of the knee joint at an early stage. Some of these abnormalities may have etiologic implications, but others may represent secondary changes developed in part as a compensatory mechanism in knee OA. PMID- 12553512 TI - Mechanoreceptors and length of the patellar ligament after Ho-YAG laser treatment: a long-term follow-up in rabbits. AB - We studied the shortening and the number of mechanoreceptors in the patellar ligament up to 18 months after Ho-YAG irradiation of the ligament's surface in 35 rabbits. The ligaments shortened an average 13% immediately after irradiation. After treatment, we divided the rabbits into a mobilized or immobilized group. At 2 weeks and 12 months after treatment both groups showed no shortening of the ligament, as compared to the intact ligament while that in the mobilized group had elongated at 12 months. Fewer Pacinian and Ruffini corpuscles were found in the irradiated ligaments than in the intact ones at 2 weeks after treatment, but we found no difference between irradiated and intact ligaments at 18 months after treatment. PMID- 12553513 TI - Increased expression of matrix metalloproteinase 1 (MMP1) in 11 patients with patellar tendinosis. AB - We studied the expression of procollagen type I, matrix metalloproteinase 1 (MMP1) and tissue inhibitor of metalloproteinase 1 (TIMP-1) by immunohistochemistry in human patellar tendinosis tissues and healthy patellar tendons. In situ gelatin zymography was used to detect collagenolytic activities. The productions of MMP1, TIMP1 and gelatinolytic activities were compared in cell cultures from tendinosis samples and controls. Tendinosis tissues and cultures showed an increase in the expression level of MMP1 and a decrease in that of TIMP1, a condition favoring collagen degradation. Gelatinolytic activities in tendinosis tissues and cultures were elevated. Collagenolysis is a striking feature in patellar tendinosis. PMID- 12553514 TI - Tumors of the head of the fibula: good function after resection without ligament reconstruction in 6 patients. AB - Tumors of the proximal fibula are rare. Wide resection includes adjacent knee stabilizing structures. Unlike treatment of traumatic disruption of the lateral ligamentous structures of the knee, wide resection of the fibular head for a tumor may not require reconstruction. After 14 years of follow-up, function was satisfactory in 6 consecutive cases where resection of the fibular head was done without ligamentous reconstruction. PMID- 12553515 TI - Clinical diagnosis of syndesmotic ankle instability: evaluation of stress tests behind the curtains. AB - We studied the feasibility of clinical tests in the diagnosis of syndesmotic injury of the ankle. 9 investigators examined 12 persons twice, including 2 patients with an arthroscopically-confirmed syndesmotic injury. They sat behind a curtain that exposed only the lower legs. We found a statistically significant relation between the final arthroscopic diagnosis and the squeeze, fibula translation, Cotton, and external rotation tests as well as for limited dorsal flexion. None of the syndesmotic tests was uniformly positive in chronic syndesmotic injury. The external rotation test had the fewest false-positive results, the fibula translation test the most. The external rotation test had the smallest inter-observer variance. The physical diagnosis was missed in one fifth of all examinations. When in accordance with medical history and physical examination, positive stress tests should raise a high index of suspicion of syndesmotic instability. The final diagnosis of such instability, however, should be made by additional diagnostic imaging and/or arthroscopy. PMID- 12553516 TI - Intra- and interobserver repeatability of radiographic measurements in hallux surgery: improvement and validation of a method. AB - To test the hypothesis that the reproducibility of radiographic measurements of the first metatarsophalangeal angle and the intermetatarsal angle I-II can be increased by exact guidelines, we calculated the intra- and interobserver reliability of both methods. 4 independent observers (2 senior residents and 2 orthopedic trainees) evaluated 50 pre- and 50 postoperative plain dorsoplantar radiographs with their method of preference and then with Mitchell et al.'s method (1958). The mean intraobserver coefficient of repeatability for the metatarsophalangeal angle improved from 5.9 degrees to 4.2 degrees and for the intermetatarsal angle I-II, from 4.4 degrees to 2.8 degrees. The interobserver coefficient of repeatability improved from 6.5 degrees to 5.0 degrees for the metatarsophalangeal angle, and from 4.9 degrees to 3.6 degrees for the intermetatarsal angle I-II. This improvement in measurement accuracy was more marked for postoperative measurements, due to deformation of the metatarsal after the osteotomy which made it more difficult to find the longitudinal axis of the metatarsal. The improvements in the accuracy of measurements were also greater in the two inexperienced observers, since their measurements differed more when they had no exact guidelines for their drawings. PMID- 12553517 TI - Abductor pollicis longus tendon interposition for arthrosis in the first carpometacarpal joint: 55 thumbs reviewed after 3 (1-5) years. AB - We have performed an interposition arthroplasty using the abductor pollicis longus (APL) tendon for arthrosis in the basal joint of the thumb in our department since 1995. Before this, we had been using an interposition with the flexor carpi radialis (FCR) tendon. We re-examined 47 patients (55 thumbs) after a median follow-up of 41 (16-60) months. The pain relief was excellent in 32 thumbs, and 25 patients improved their ability to perform daily tasks. Mobility was well preserved. Key pinch and grip strengths averaged 78% and 89%, respectively, of those in unaffected hands. An excellent general result was reported by 34 patients, while 4 patients would not have consented to the operation if they had known the outcome in advance. PMID- 12553518 TI - Bacterial contamination in postmortem bone donors. AB - We analyzed factors influencing the contamination rate of allografts and blood samples obtained from postmortem bone donors. 5,710 allografts were harvested, from 550 donors of which 3,164 (55%) were swab culture negative. Blood cultures were positive in 140 donors (26%). The risk of graft contamination increased with each extra team member (Odds Ratio 1.9). It was also higher with organisms of greater virulence in donors with a positive blood culture (OR 3.5). The risk of blood contamination increased per hour postmortem (OR 1.1) and the same increase was seen with organisms of high virulence. In donors with multiple trauma, the risk of blood contamination with organisms of high virulence was greater (OR 8.2), but smaller in donors with preceding organ procurement (OR 0.1). To minimize the bacterial load, donors should be obtained in operating rooms, using aseptic techniques with only a few personnel for procurement. The postmortem time should be kept to a minimum. The procurement cultures from donors with multiple trauma should be carefully interpreted. Blood cultures should be taken into account, since these can help to find contamination not detected by swab cultures. PMID- 12553519 TI - The dynamic volume changes of polymerising polymethyl methacrylate bone cement. AB - The Swedish hip register found an increased risk of early revision of vacuum mixed cemented total hip replacements. The influence of cement mixing technique on the dynamic volume change in polymerising PMMA is not well understood and may be relevant to this observation. Applying Archimedes' principle, we have investigated the dynamic volume changes in polymerising cement and determined the influence of mixing technique. All specimens showed an overall volume reduction: hand-mixed 3.4% and vacuum-mixed 6.0%. Regression analysis of sectional porosity and volume reduction showed a highly significant relationship. Hand-mixed porous cement showed a transient volume increase before solidification. However, vacuum mixed cement showed a progressive volume reduction throughout polymerisation. Transient expansion of porous cement occurs at the critical time of micro interlock formation, possibly improving fixation. Conversely, progressive volume reduction of vacuum-mixed cement throughout the formation of interlock may damage fixation. Stable fixation of vacuum-mixed cement may depend on additional techniques to offset the altered volumetric behaviour of vacuum-mixed cement. PMID- 12553520 TI - Mechanical characteristics of antibiotic-laden bone cement. AB - We studied the mechanical characteristics of cement-antibiotic combinations in vitro. Palacos R was tested without antibiotics, with gentamicin alone and with gentamicin plus vancomycin or flucloxacillin. Palacos LV was studied only with gentamicin added. CMW 1 was studied with gentamicin added, with gentamicin plus vancomycin, and with gentamicin plus flucloxacillin. We performed four-point bending tests on beams of cement to establish bending strength and modulus, and compared the values to ISO standards. Density was also assessed. Palacos R was the strongest of the cements (bending strength 80 MPa). Palacos formulations (apart from Palacos LV) had a higher density and bending modulus than CMW 1. Statistical comparison of various cements with plain Palacos R showed lower density in 4 of the mixtures, and lower bending strength and modulus in 6 of the mixtures. Palacos R/gentamicin plus vancomycin and CMW 1/gentamicin plus vancomycin had bending strength slightly above minimum ISO standards, suggesting that the addition of vancomycin during cement mixing may compromise the outcome in revision surgery for sepsis. PMID- 12553521 TI - Does exercise reduce the burden of fractures? A review. AB - The null hypothesis that exercise has no effect on fracture rates in old age cannot be rejected on the basis of any published, randomized, prospective data. The view that exercise reduces the number of fractures is based on prospective and retrospective, observational cohort studies and case-control studies, all hypothesis-generating, not hypothesis-testing. Consistently replicated sampling bias may confirm the finding when evaluating other than randomized prospective studies. Better health, better muscle function, more muscle mass, better coordination may lead to exercise. The causal relationship could be between better health and exercise and better health and fewer fractures, not exercise and fewer fractures. The hypothesis should be tested in prospective, randomized studies evaluating hip, spine and other fragility fractures separately. Blinded studies assessing the effects of exercise can obviously not be done, but open trials can and should be undertaken to increase the level of evidence within the evidence-based system. There are firm data supporting the view that exercise during growth builds a stronger skeleton. Exercise during growth seems to result in high peak BMD and high muscle strength. However, the Achilles heel of exercise is its cessation. Are the skeletal and muscular benefits attained during growth retained after the cessation of exercise and can any residual benefits be found in old age, the period when fragility fractures rise exponentially? Does exercise during adulthood produce any biologically important reduction in surrogate end points for fractures other than BMD, since BMD can be influenced only marginally by exercise after completion of growth? Recommendations for exercise should be based on evidence, not on opinion. Can continued recreational exercise maintain some of the benefits in BMD and muscle function achieved in youth? What level of recreational exercise is needed to retain these benefits, if not fully, then at least to some extent? Dose-response relationships should be quantified. Furthermore, the effect of exercise on independent, surrogate end-points for fractures, such as bone size, shape, architecture, muscle function, fall frequency and frequency of injurious falls during defined periods in the life cycle must be determined. Absence of evidence is not evidence of absence of effect, but if we recommend exercise then should this be to children, adults, elderly, men and women with fractures, all persons? What type of exercise? For how long? Lifelong? If exercise could be implemented for most persons in society, would this reduce the number of fractures? Would the increased costs associated with the efforts to increase the activity level be lower than the reduced costs associated with any reduction in fractures? Our inability to answer these questions must be acknowledged before recommendations are made at the community level. PMID- 12553522 TI - Retrograde intramedullary nailing of distal humeral nonunions: a new technique with good results in 5 patients. PMID- 12553523 TI - Rib bone graft for posterior spinal fusion in children. AB - Treatment of columnar instability in children with conventional fusion procedures may preclude normal growth and cause dysfunction and pain due to malalignment or reduced mobility. To achieve normal growth, we have treated spinal instability in 7 children with posterior fusion, using bilateral rib transplants secured by horizontal laminar cerclages. Solid fusion was obtained and no serious complications occurred. PMID- 12553524 TI - Simultaneous leg lengthening and knee arthrodesis using an external ring fixator- a case report. PMID- 12553525 TI - Fracture of the index sesamoid--a case report. PMID- 12553526 TI - Extending our understanding of burnout: test of an integrated model in nonservice occupations. AB - C. L. Cordes and T. M. Dougherty (1993) provided a conceptual framework of job burnout in nonservice organizations. This study sought to determine the "fit" of that theoretical model within nonservice occupations. LISREL VIII was used to test this model on 165 participants, and the overall model fit the data well. Supervisory support moderated the relationships between the role conflict, role ambiguity, and quantitative role overload stressors and emotional exhaustion and between emotional exhaustion and depersonalization. Unexpectedly, role conflict, role ambiguity, and quantitative role overload had a positive impact on emotional exhaustion when supervisory support was high. As expected, participation had a negative association with depersonalization. Employees experiencing emotional exhaustion were more likely to feel nervous or tense at work. PMID- 12553527 TI - Personality, work characteristics, and employee well-being: a longitudinal analysis of additive and moderating effects. AB - This study tested the longitudinal influence of personality (measured by the characteristics growth need strength, negative affectivity [NA], and upward striving) on 3 psychological outcomes (intrinsic work motivation, emotional exhaustion, and turnover intention), using a pattern of specific relationships between work characteristics and these outcomes as a framework. The study hypotheses were tested in a multioccupational sample consisting of bank employees and teachers, using a 2-wave panel design with a 1-year time interval and structural equation modeling. NA had a cross-lagged direct and additive relationship with emotional exhaustion and also moderated the relationship between Time 1 workload and Time 2 emotional exhaustion. The authors concluded that NA may have multiple effects on emotional exhaustion that persist over time. PMID- 12553528 TI - Predictors of overall and on-the-job substance use among young workers. AB - Predictors of overall and on-the-job substance (alcohol and marijuana) use were studied in a sample of young workers. The 18 predictors represent 6 general domains of risk factors: demographic, personality, substance use outcome expectancies, workplace substance availability, workplace social control, and work stressors. Data were obtained from a sample of 319 individuals ages 16 to 19. Hierarchical tobit regression analyses revealed that 5 of the 6 domains of risk factors were related to employee substance use. Similarities and differences were found in the predictors of overall and on-the-job substance use and in the predictors of alcohol and marijuana use. PMID- 12553529 TI - Emotion regulation in customer service roles: testing a model of emotional labor. AB - The study used a time-sampling method to test aspects of A. Grandey's (2000) emotion regulation model of emotional labor. Eighteen customer service employees from a call center recorded data on pocket computers every 2 hr at work for 2 weeks. Participants completed ratings of emotion regulation, events, expressed and felt emotions, well-being, and performance on 537 occasions and completed questionnaires containing individual and organizational measures. Multilevel analyses supported many aspects of the model but indicated that it has to be implemented precisely in terms of regulating emotion for organizational goals. Results also showed that deep and surface acting had different consequences for employees. Overall, the study found that emotion regulation is a viable platform for understanding emotional labor. PMID- 12553530 TI - Accidental outcomes: attitudinal consequences of workplace injuries. AB - A model of the attitudinal outcomes of the occurrence and severity of occupational injuries was developed and tested. The model postulates that workplace accidents result in a perceived lack of influence and a distrust of management, with the former also affecting the distrust of management. Both are hypothesized to predict job dissatisfaction. Exit (turnover intentions) and voice (perceptions of union instrumentality) are hypothesized as outcomes of job dissatisfaction. A sample of 9,908 employees was tested with the 1995 Australian Workplace Industrial Relations Survey database. Structural equation modeling provided strong support for the model with respect to accident occurrence, and the model was replicated across 8 different occupational groups. There was less support for the model with respect to accident severity. PMID- 12553531 TI - Advances in gene therapy for bladder cancer. AB - The efficacy of various currcntly available therapeutic strategies for bladder cancer is not always suffcient, especially for the advanced disease, recurrent superficial cancer, and treatment-resistant carcinoma in situ. Advances in genetic and molecular biology have led to novel approaches for cancer treatment. Gene therapy is currently one of the most promising strategies against various malignancies, and several clinical trials have been approved worldwide. Various strategies for modulating the genetic state have been applied in bladder cancer treatment, and encouraging results have been demonstrated both in vitro and in vivo. Although the therapeutic genes work dramatically when the transgenes are effectively expressed in the targeted cells, however, a sufficient rate of transduction cannot always be achieved. The most significant obstacle for clinical application of cancer gene therapy might be the method for sufficient delivery and expression of the therapeutic genes. Bladder is an easily accessible organ because of its anatomy; however, a glycosaminoglycan (GAG) layer on the bladder mucosa may protect integration of exo-delivered genetic vectors. Various strategies are applied for improving the transduction efficacy of the therapeutic genes into the bladder cancer cells. These strategies include the modification of adenoviral fibers, cotransduction of the materials for enhancing the viral infectivity, and disruption of the GAG layer. Recent advances in the field of gene therapy for bladder cancer are briefly summarized in this review. PMID- 12553532 TI - Suicide gene therapy mediated by the Herpes Simplex virus thymidine kinase gene/Ganciclovir system: fifteen years of application. AB - Gene-directed enzyme prodrug therapy (GDEPT) is a two step therapeutic approach for cancer gene therapy. In the first step, the transgene is delivered into the tumor and expressed. In the second step a prodrug is administered and is selectively activated by the expressed enzyme. The first GDEPT system described was the thymidine kinase gene of the Herpes Simplex virus (HSVtk) in combination with the prodrug Ganciclovir (GCV). A large number of experiments have been performed with this system, in different types of tumors and initial studies in animal models were very promising. This encouraged investigators to move into clinical trials although poor results have been obtained so far. A large effort has been made with numerous different strategies to enhance HSVtk/GCV efficacy in cellular and in vivo models and very strong cytotoxic effects have been obtained. The present review describes the current state of preclinical research and summarizes the results of the clinical trials undertaken. PMID- 12553533 TI - Development of improved factor VIII molecules and new gene transfer approaches for hemophilia A. AB - Hemophilia A, the most common inherited bleeding disorder, is caused by deficiency or functional defects in coagulation factor VIII (fVIII). Conventional treatment for this disease involves intravenous infusions of plasma-derived or recombinant fVIII products. Although replacement therapy effectively stops the bleeding episodes, it has a risk of transmission of viral blood-borne diseases and development of neutralizing antibodies that inactivate the administered fVIII protein. Hemophilia A is an attractive candidate for application of gene therapy approaches because the therapeutic window is wide and even modest elevation of fVIII levels will correct the hemophilic phenotype. Ongoing preclinical investigations utilize animal models of hemophilia A, including genetically fVIII deficient mice and naturally fVIII-deficient dogs, to optimize vectors, transgenes and target cell populations for Phase I clinical trials. In this review, we outline the progress in understanding the mechanisms of fVIII turnover, which provides a basis for development of improved fVIII molecules with prolonged half-life in the circulation. We discuss the possibility of incorporating these improved fVIII molecules as transgenes into self-inactivating lentiviral vectors carrying chromatin insulator sequences, representing a new generation of gene delivery vehicle, to target hematopoietic stem cells and endothelial cells. The use of hematopoietic stem cells as the target cell population may prevent inhibitor formation to transduced fVIII by induction of immune tolerance. Alternatively, endothelial cells may support optimal synthesis of fVIII and myeloablative conditioning of patients with radiation or chemotherapy may not be required for efficient engraftment of the engineered cells. Collectively, these proposed advances represent promising prophylactic strategies toward long-term correction of the coagulation defect in this progressively debilitating, life-threatening disease. PMID- 12553534 TI - Recent advances in adenovirus-mediated gene therapy for cerebral ischemia. AB - Cerebral ischemia induces many degenerative cellular reactions, including the release of excitatory amino acids, the formation of oxygen free radicals, Ca2+ overload, the activation of several cellular enzyme systems such as Ca2+ dependent proteases, and the initiation or genomic responses that can affect the tissue outside the area of reduced blood flow. Furthermore, increasing evidence indicates that apoptosis contributes to the death of brain cells following cerebral ischemia. Several studies have shown that cerebral ischemia alters the expression of genes, some of which may play protective or harmful roles. Although many genes have the potential to treat cerebral ischemia, target genes or their translated products are often difficult to express, if at all, in brain cells. However, adenovirus-mediated gene transfer can overcome this disadvantage. To date, many treatment strategies have been developed for cerebral ischemia using target genes such as neuronal apoptosis inhibitory protein (NAIP), glial cell line-derived neurotrophic factor (GDNF), sensitive to apoptosis gene (SAG), 150 kDa oxygen-regulated protein (ORP150), etc. Moreover, new vectors and gene delivery systems are constantly being invented although there is no perfect vector to date. Gene therapy could constitute a powerful strategy to treat cerebral ischemia in the near future. PMID- 12553535 TI - Gene therapy for ischemic brain diseases. AB - We discuss possible gene therapies for the treatment of ischemic diseases in the central nervous system (CNS). These therapies aim at the prevention of carotid artery restenosis, stimulation of angiogenesis for ischemic brain, protection of neurons against ischemia, and prevention of vasospasm due to subarachnoid hemorrhage (SAH). Carotid artery restenosis can perhaps be approached by preventing vascular smooth muscle cell proliferation via gene therapy in addition to surgical treatment. Cerebral angiogenesis therapy might be applicable to moyamoya disease. Gene therapies with VEGF and HGF to stimulate angiogenesis have been successful in muscle; however, efficacy in the CNS is unknown. Gene transfection efficiency of viral vectors has been poor in the CNS, and the safety of such vectors is questionable. Therefore, development of gene therapy is for neural protection and prevention of vasospasm due to SAH has been limited. Infusion of HVJ-AVE liposomes into monkey cerebrospinal fluid (CSF) space yielded wide-spread gene transfection. HVJ-AVE liposomes may be a promising vector for use in the human CNS. Few currently available gene therapies appear to be options for clinical treatment of cerebral ischemia despite many experimental designs. In addition to the inherent difficulties of treating the CNS, vectors and methods for introducing vectors into the CNS must be improved. PMID- 12553536 TI - Application of in vivo electroporation to cancer gene therapy. AB - Much intensive research has gone into the development of safe and efficient methods for the delivery of therapeutic genes. In vivo electroporation is a non viral delivery protocol in which plasmid DNA solutions are injected into targeted tissues, followed by electric pulses (typically 100 V, 50 ms). In general, in vivo electroporation enhances gene expression in targeted tissues by 2-3 orders of magnitude, as compared to the injection of plasmid DNA solutions without electric pulses, and the tissue damage appears to be minimal. Among the other advantages of this technique are that it can safely be administered repeatedly, and it is simpler and more economical to use than viral vectors, especially in clinical cases. Using this approach, highly efficient gene transfer has already been achieved in muscle and liver as well as in tumors. In fact, gene therapies for cancer utilizing in vivo electroporation have been proved effective in a number of experimental murine tumor models. The therapeutic genes delivered in those cases were diverse including, for example, cytokine genes (IL-12) and cytotoxic genes (TRAIL), making possible a wide range of therapeutic strategies. Moreover, systemic antitumor effects were also observed, suggesting that this approach may be effective for the treatment of metastatic as well as primary tumors. PMID- 12553537 TI - Diabetes gene therapy: potential and challenges. AB - Gene therapy, developing rapidly as a result of advances in molecular biology and the Human Genome Project, is now highlighted as a most hopeful technology of the 21st century. The major goal of gene therapy in diabetes mellitus (DM) is to maintain euglycemia in face of wide variations in dietary intake. Although some obstacles remain to be overcome, the risk-benefit ratio of gene therapy in DM is better than that of lifelong injections of insulin, and islet transplantation, which faces the problems of donor shortage and rejection. This review focuses on the recent advances in gene therapy of insulin-requiring diabetes, with particular emphasis on 1. the gene delivery systems by viral vectors, since most gene therapy approaches for DM involve the use of viral vectors, paying special attention to current efforts to overcome the disadvantages of adenovirus, adenovirus-associated virus and retrovirus vectors and targeting gene delivery for optimal efficiency of gene expression; 2. coupling the synthesis and release of the transgene insulin to serum glucose concentrations, especially with reference to the current promoters controlling at transcriptional level the ectopic insulin expression in autologous hepatocytes; 3. beta-cell replacement strategies: engineering of beta-cells, especially those derived from pluripotent stem cells, non beta-cells, and on a new comer, the K cells. Recent advances in the use of stem cells for potential application in diabetes gene therapy are also discussed. PMID- 12553538 TI - Aberrometry: the past, present, and future of optometry. PMID- 12553539 TI - Measurement of refractive errors in young myopes using the COAS Shack-Hartmann aberrometer. AB - PURPOSE: To evaluate the Complete Ophthalmic Analysis System (COAS; WaveFront Science) for accuracy, repeatability, and instrument myopia when measuring myopic refractive errors. METHODS: We measured the refractive errors of 20 myopic subjects (+0.25 to -10 D sphere; 0 to -1.75 D cylinder) with a COAS, a phoropter, and a Nidek ARK-2000 autorefractor. Measurements were made for right and left eyes, with and without cycloplegia, and data were analyzed for large and small pupils. We used the phoropter refraction as our estimate of the true refractive error, so accuracy was defined as the difference between phoropter refraction and that of the COAS and autorefractor. Differences and means were computed using power vectors, and accuracy was summarized in terms of mean vector and mean spherocylindrical power errors. To assess repeatability, we computed the mean vector deviation for each of five measurements from the mean power vector and computed a coefficient of repeatability. Instrument myopia was defined as the difference between cycloplegic and noncycloplegic refractions for the same eyes. RESULTS: Without cycloplegia, both the COAS and autorefractor had mean power vector errors of 0.3 to 0.4 D. Cycloplegia improved autorefractor accuracy by 0.1 D, but COAS accuracy remained the same. For large pupils, COAS accuracy was best when Zernike mode Z4(0) (primary spherical aberration) was included in the computation of sphere power. COAS repeatability was slightly better than autorefraction repeatability. Mean instrument myopia for the COAS was not significantly different from zero. CONCLUSIONS: When measuring myopes, COAS accuracy, repeatability, and instrument myopia were similar to those of the autorefractor. Error margins for both were better than the accuracy of subjective refraction. We conclude that in addition to its capability to measure higher order aberrations, the COAS can be used as a reliable, accurate autorefractor. PMID- 12553540 TI - Comparison of monochromatic ocular aberrations measured with an objective cross cylinder aberroscope and a Shack-Hartmann aberrometer. AB - Repeated measures of wavefront aberrations were taken along the line-of-sight of seven eyes using two instruments: an objective, cross-cylinder aberroscope (OA) and a Shack-Hartmann (SH) aberrometer. Both instruments were implemented on the same optical table to facilitate interleaved measurements on the same eyes under similar experimental conditions. Variability of repeated measures of individual coefficients tended to be much greater for OA data than for SH data. Although Zernike coefficients obtained from a single measurement were generally larger when measured with the OA than with the SH, the averages across five trials were often smaller for the OA. The Zernike coefficients obtained from the two instruments were not significantly correlated. Radial modulation-transfer functions and point-spread functions derived from the two sets of measurements were similar for some subjects, but not all. When average Zernike coefficients were used to determine optical quality, the OA indicated superior optics in some eyes, but the reverse trend was true if Zernike coefficients from individual trials were used. Possible reasons for discrepancies between the OA and SH measurements include difference in sampling density, quality of data images, alignment errors, and temporal fluctuations. Multivariate statistical analysis indicated that the SH aberrometer discriminated between subjects much better than did the objective aberroscope. PMID- 12553541 TI - Aberrations of the human eye in visible and near infrared illumination. AB - PURPOSE: In most current aberrometers, near infrared light is used to measure ocular aberrations, whereas in some applications, optical aberration data in the visible range are required. We compared optical aberration measurements using infrared (787 nm) and visible light (543 nm) in a heterogeneous group of subjects to assess whether aberrations are similar in both wavelengths and to estimate experimentally the ocular chromatic focus shift. METHODS: Ocular aberrations were measured in near infrared and visible light using two different laboratory developed systems: laser ray tracing (LRT) and Shack-Hartmann. Measurements were conducted on 36 eyes (25 and 11 eyes, respectively), within a wide range of ages (20 to 71 years), refractive errors (-6.00 to +16.50), and optical quality (root mean square wavefront error, excluding defocus, from 0.40 to 9.89 microm). In both systems, wave aberrations were computed from the ray aberrations by modal fitting to a Zernike polynomial base (up to seventh order in laser ray tracing and sixth order in Shack-Hartmann). We compared the Zernike coefficients and the root mean square wavefront error corresponding to different terms between infrared and green illumination. RESULTS: A Student's t-test performed on the Zernike coefficients indicates that defocus was significantly different in all of the subjects but one. Average focus shift found between 787 nm and 543 nm was 0.72 D. A very small percentage of the remaining coefficients was found to be significantly different: 4.7% of the 825 coefficients (25 eyes with 33 terms) for laser ray tracing and 18.2% of the 275 coefficients (11 eyes with 25 terms) for Shack-Hartmann. Astigmatism was statistically different in 8.3% of the eyes, root mean square wavefront error for third-order aberrations in 16.6%, and spherical aberration (Z4(0)) in 11.1%. CONCLUSIONS: Aerial images captured using infrared and green light showed noticeable differences. Apart from defocus, this did not affect centroid computations because within the variability of the techniques, estimates of aberrations with infrared were equivalent to those measured with green. In normal eyes, the Longitudinal Chromatic Aberration of the Indiana Chromatic Eye Model can predict the defocus term changes measured experimentally, although the intersubject variability could not be neglected. The largest deviations from the prediction were found on an aphakic eye and on the oldest subject. PMID- 12553542 TI - A method to predict refractive errors from wave aberration data. AB - We explored the impact of the eye's higher-order aberrations on subjective refraction comparing two classes of methods for estimating refractive state, one based directly on the wave aberration defined in the pupil plane and another based on the retinal image plane. The method defined in the pupil plane chose the sphere and cylinder that either minimized the wave aberration root mean square or minimized the sum of all the spherical and cylindrical components in the wave aberration. The method defined in the image plane chose the sphere and cylinder that optimized an image-quality metric such as the Strehl intensity ratio, the entropy and the intensity variance of the point-spread function, the volume under the modulation transfer function, or the volume under the contrast-sensitivity function. All these methods were compared in a population of six eyes for which we measured both the wave aberration with a Shack-Hartmann wavefront sensor and the subjective refraction under identical conditions. Pupil plane methods predicted subjective refraction poorly. The mean absolute error of the prediction, in spherical equivalent, was about 0.5 D (range, 0.1 to 0.8 D) and increased with increases in higher-order aberrations. However, for all the retinal image plane methods, the mean error between predicted and subjective refraction was about 0.1 D (range, 0 to 0.25 D). The reliability of the method based on the image-quality optimization was further confirmed in a large population of 146 eyes. In conclusion, higher-order aberrations influence the amount of sphere and cylinder required to correct vision. The results indicate that subjective refraction can be predicted from the eye's optics alone by optimizing computed retinal image quality. PMID- 12553543 TI - Relationship between refractive error and monochromatic aberrations of the eye. AB - PURPOSE: To examine the relationship between ametropia and optical aberrations in a population of 200 normal human eyes with refractive errors spanning the range from +5.00 to -10.00 D. METHODS: Using a reduced-eye model of ametropia, we tested the hypothesis that the optical system of the eye is uncorrelated with the degree of ametropia. These predictions were evaluated experimentally with a Shack Hartmann aberrometer that measured the monochromatic aberrations across the central 6 mm of the dilated pupil in well-corrected, cyclopleged eyes. RESULTS: Optical theory predicted, and control experiments on a model eye verified, that Shack-Hartmann measurements of spherical aberration will vary with axial elongation of the eye even if the dioptric components of the eye are fixed. Contrary to these predictions, spherical aberration was not significantly different from emmetropic eyes. Root mean square of third-order aberrations, fourth-order aberrations, and total higher aberrations (third to 10th) in myopic and hyperopic eyes were also uncorrelated with refractive error. Astigmatic eyes tended to have larger total higher-order aberrations than nonastigmatic eyes. CONCLUSIONS: We conclude that a reduced-eye model of myopia assuming fixed optical parameters and variable axial length is not tenable. PMID- 12553544 TI - Double-pass measurement of retinal image quality in the chicken eye. AB - PURPOSE: The chicken, Gallus gallus domesticus, is used as an animal model to study the development of refractive error. Although vision is important in determining the eye's refractive state, relatively little is known about the retinal image quality of the chicken eye. An objective double-pass technique was used to measure the optical quality of the eyes of White Leghorn chickens. METHODS: Measurements were made on 21 eyes of six untreated birds and eight experimental birds that were members of a study of refractive development. Ages ranged from 3 to 6 weeks, and refractions ranged from -1.29 to +0.58 D in the untreated eyes and -4.58 to +10.17 D in the experimental eyes. The measurements were made under general anesthesia combined with either cycloplegia or ciliary nerve section. Proper optical alignment of the eye was achieved with the aid of a TV monitor, CCD camera, and an infrared source. A 543-nm laser point source was focused on the retina, and the double-pass aerial image was collected by a high resolution CCD camera. Refractive errors were corrected with trial lenses, using a bracketing method to optimize the retinal images. Both the full width at half maximum of the double-pass aerial image and the single-pass modulation transfer function were used as objective estimates of the optical quality. RESULTS: The mean full width at half-maximum value in eyes of the untreated birds was 1.60 min arc for a 4.50-mm mean pupil diameter. Optical quality tended to be worse in the experimental myopic eyes. CONCLUSIONS: The optical quality of the chicken eye measured under monochromatic conditions meets or may even exceed the neural limits of spatial acuity based on anatomical estimates of ganglion cell spacing. The data also suggest that optical quality is worse in myopic eyes, which is consistent with studies of human eyes. PMID- 12553545 TI - Scatter and its implications for the measurement of optical image quality in human eyes. AB - PURPOSE: To investigate the effect of scatter on measurements of wavefront aberrations and point-spread functions in a model eye. METHODS: The wavefront aberrations of a model eye were measured using Hartmann-Shack wavefront sensing and crossed-cylinder aberroscope techniques and compared with its measured point spread function in the presence of scattering media of different concentrations. RESULTS: The point-spread functions became broader as the concentration increased. Forward light scatter on both the light path into the eye and the light path out of the eye contributed to this broadening of the point-spread function. Neither the crossed-cylinder aberroscope nor wavefront sensing, which, respectively, measure the ocular wavefront aberrations for light entering the eye and leaving the eye, were affected by the scatter. CONCLUSION: We predict that by minimizing the contribution of the forward light scatter from one or other of these light paths by manipulating the size of the entrance and exit pupils, it should be possible to objectively assess narrow-angle forward light scatter in the eye by measuring and removing any confounding effect from wavefront aberration. PMID- 12553546 TI - Use of retroillumination to visualize optical aberrations caused by tear film break-up. AB - PURPOSE: The aim of the current study was to develop quantitative methods to assess optical aberrations caused by tear film disruption. METHODS: We used standard fluorescein imaging (FL) and a novel retroillumination (RI) method to image tear film disruption in 12 eyes. Using a clinical slit lamp biomicroscope, we alternated between widefield blue and narrow-beam white light to obtain an interleaved series of FL and RI images of the time course and pattern of tear film break-up. We developed an optical analysis that indicates that the RI image should be proportional to the spatial derivative of the FL image. Intensity fluctuations in the RI images are due to thickness changes in the tear film, whereas intensity fluctuations in FL images are directly determined by tear film thickness. RESULTS: As predicted by optical analysis of RI, the spatial distribution of gaps in the tear film seen with fluorescein appeared as pairs of light and dark contours in the RI images, and a precise correspondence between the spatial derivative of the FL image (slope) and the RI image was found. Both methods showed a gradual spreading of the tear disruption during blink suppression that varied tremendously among eyes in both time and spatial pattern. Resumption of normal blinking did not produce an immediate reconstitution of the normal tear film, and areas of tear break-up created during blink suppression remained abnormal for up to several minutes of normal blinking. CONCLUSIONS: Our analysis indicates that both FL and RI have the potential to quantify optical changes occurring during tear break-up. These results support an interpretation of RI as an intensity-based method for mapping the highly irregular optical aberrations of the eye produced by tear film disruption. PMID- 12553547 TI - A new method for describing the aberrations of the eye using Zernike polynomials. AB - The standard Zernike polynomial functions are reformulated in a way so that the number of functions (or terms) needed to describe an arbitrary wavefront surface to a given Zernike radial order is reduced by a factor of approximately two, and the terms are described in a fashion quite similar to that used to describe common sphero-cylindrical errors of the eye. A wavefront is represented using these terms by assigning a pair of values, a magnitude and an axis, to all terms that are radially symmetric so that the individual aberrations are presented in a way similar to the way common astigmatism is currently given in terms of cylinder power and axis. The root mean square of these magnitudes gives the root mean square wavefront error just as does the root mean square of the standard Zernike coefficients. Formulas are given to convert standard Zernike coefficients to the magnitude and axis values. PMID- 12553548 TI - Autonomic dysfunction associates with prolongation of QT intervals and blunted night BP in obese women with visceral obesity. AB - Prolonged QT intervals and a reduced fall of nocturnal blood pressure (BP) both predict an increased risk of cardiovascular events in obese subjects. We evaluated circadian BP variations (24-h ambulatory BP monitoring), autonomic function (power spectral analysis of RR interval oscillations) and cardiac repolarization times (QTc-dispersion and QTc interval) in 70 obese women, aged 25 44 yr, grouped by WHR into group A (WHR > 0.85, no.=38) and group B (WHR < or = 0.85, no.=32). Compared with non-obese age-matched women (no.=25, BMI=23+/-1.8) and obese women of group B, obese women of group A had higher values of QTc-d (p<0.05) and QTc (p<0.05), an altered sympathovagal balance (ratio of low frequency/high-frequency power, p<0.01), and a blunted nocturnal drop in BP (p<0.01). In group A, QTc-d and the QTc interval correlated with diastolic night BP (p<0.01) and sympathovagal balance (p<0.01). WHR and plasma insulin levels correlated with QT intervals, reduced nocturnal fall in diastolic BP and sympathovagal balance (p<0.01). Prolongation of cardiac repolarization times and the reduction of nocturnal fall in BP coexist in obese women with visceral obesity, and might contribute to their raised cardiovascular risk. Autonomic dysfunction may be the common mechanism for this association. PMID- 12553549 TI - Euglycemic hyperinsulinemia, but not lipid infusion, decreases circulating ghrelin levels in humans. AB - The orexigenic and anabolic gastric hormone ghrelin is secreted in response to acute and chronic energy requirements. While pre-prandial increases and post prandial decreases of plasma ghrelin levels in rodents and humans seem to indicate a role for the novel peptide hormone as an afferent meal initiator or "hunger hormone", the precise mechanisms which are suppressing ghrelin secretion in response to caloric intake remain largely unknown. We show here that human ghrelin levels decrease by almost 50% under hyperinsulinemic euglycemic clamp conditions (no.=4, p=0.001), revealing physiologically relevant increases of insulin levels as an independent determinant of circulating ghrelin levels. In a second study, 3-4-fold increased plasma free fatty acid levels, as another metabolic candidate for the modulation of circulating ghrelin concentrations, were generated by constant lipid infusion, but failed to change plasma ghrelin. Simultaneous elevation of free fatty acids and insulin again markedly decreased ghrelin concentration (no.=4, p=0.01). Insulin induced suppression of circulating ghrelin levels (or the lack thereof) could be a mechanism with relevance for the understanding of the (patho-) physiology of meal initiation and termination, the pathogenesis of the metabolic syndrome and for the development of respective therapeutic perspectives. PMID- 12553550 TI - Acromegaly and thyroid. PMID- 12553551 TI - Hypothalamus-pituitary-adrenal axis in central diabetes insipidus: ACTH and cortisol responsiveness to CRH administration. AB - A strong relationship has been found between arginine-vasopressin (AVP) and hypothalamus-pituitary-adrenal axis in humans. The aim of the current study was to evaluate baseline and CRH-stimulated ACTH and F levels in patients with central diabetes insipidus (CDI), before and after replacement therapy with desamino-D-AVP (DDAVP). Twenty-five patients with CDI, and 25 sex- and age- and BMI-matched healthy subjects entered the study. A standard CRH test (measurement of plasma ACTH and serum F before and every 15 min for 2 h after the administration of 100 microg of human CRH) was performed in all subjects. In patients with CDI, CRH test were repeated after 1 week of DDAVP at standard doses. At study entry, ACTH and F levels were significantly higher in patients with CDI than in controls either at baseline (ACTH: 45.5+/-4.8 vs 18.5+/-3.3 ng/l, p<0.05; F: 375.1+/-55.7 vs 146.6+/-19.4 microg/l, p<0.05) or after CRH test considered as a peak (ACTH: 90.8+/-14.4 vs 42.5+/-7.4 ng/l, p<0.05; F: 501.6+/ 65.7 vs 226.3+/- 25.6 microg/l, p<0.05) and AUC (ACTH: 3997.0+/-571.7 vs 2136.0+/ 365.8 ng/l/120 min, p<0.05; F: 31,489.0+/-4299.4 vs 14,854.5+/-1541.5 microg/l/120 min, p<0.05). In patients with CDI, 1 week of replacement with DDAVP brought down ACTH (peak: 56.9+/-9.3 ng/l; AUC: 2390.7+/-480.7 ng/l/120 min) and F (peak: 310.3+/-39.5 microg/l; AUC: 17,555.5+/-2008.7 microg/l/120 min) responses to CRH to normal but did not significantly modify baseline hormone levels (ACTH: 29.6+/-3.6 ng/l; F: 239.0+/-32.3 microg/l). In conclusion, CDI is associated to increased baseline ACTH and F levels and increased responsiveness of ACTH and F to CRH administration. In addition, replacement treatment with DDAVP normalized CRH-induced but not baseline ACTH and F secretion. PMID- 12553552 TI - Expression of endocrine autoantibodies in chronic hepatitis C, before and after interferon-alpha therapy. AB - Interferon-alpha (IFN-alpha) treatment for chronic hepatitis C (CHC) has been associated with thyroid autoimmunity and/or dysfunction. Only a few data concerning the prevalence of islet-cell or adrenal cortex autoantibodies in IFN alpha-treated subjects are currently available. The aims of our study were to evaluate in CHC, 1) the prevalence and association of thyroid, islet-cell and adrenal autoantibodies, and 2) the appearance of endocrine dysfunction, before and after a 6 month IFN-alpha treatment. We analyzed serum samples from 203 adult patients at the time of clinical diagnosis of CHC and showed that the prevalence of thyroperoxidase (TPOAb), thyroglobulin (TGAb), TSH-receptor (TRAb), glutamic acid decarboxylase (GAD65Ab), IA-2/ICA512 (IA-2/ICA512Ab) and 21-hydroxylase (21OHAb) autoantibodies was similar to that observed among healthy control subjects of similar age and sex distribution. Among 99 patients with follow-up serum samples, 83 accepted and 16 refused IFN-alpha treatment. The IFN-alpha treatment was associated with increase of TPOAb levels in 3 subjects already positive at baseline, with progression to overt hypothyroidism in 2 of them. The de novo appearance of autoantibodies was observed in 5/80 (6%) cases for TPOAb, 1/81 (1.2%) for GAD65Ab and 2/81 (2.5%) for IA-2/ICA512Ab. Clinical or subclinical signs of either hyperthyroidism or hypothyroidism were demonstrated in 3/5 cases with de novo appearance of TPOAb. Four subjects, initially positive for either GAD65Ab or IA2/ICA512Ab, were all found negative after IFN-alpha treatment. No subjects showed positivity for 21OHAb either at baseline or after the follow-up period. Our study suggests that, in CHC untreated patients, the prevalence of endocrine autoantibodies is similar to that observed in the general population. Furthermore, we demonstrate that IFN-alpha treatment is associated with the induction or enhancement of thyroid, but not of islet-cell or adrenal cortex autoimmunity. PMID- 12553553 TI - Three cases of papillary carcinoma and three of adenoma in thyroglossal duct cysts: clinical-diagnostic comparison with benign thyroglossal duct cysts. AB - The clinical and diagnostic findings of 3 cases of papillary thyroid carcinoma in thyroglossal duct cyst (TDC) were compared to those of 3 cases of adenoma in TDC and 2 cases of benign TDC. The neck masses of the subjects with benign TDC grew slowly, whereas those of 2 patients with papillary carcinoma and 1 of the patients with adenoma grew rapidly (especially those with carcinoma). On the other hand, one case of carcinoma, and two cases of adenoma in TDC were diagnosed incidentally. Benign TDC had an anechoic pattern at US, whereas the cysts containing carcinoma and adenoma showed the presence of a mural nodule at US. Microcalcifications in the mural mass were present in one patient with carcinoma. The 3 patients with carcinoma in TDC underwent total thyroidectomy. The histology was negative in all 3 patients for thyroid cancer and thyroid nodules. However, in 2 of them it revealed the carcinoma invading the cyst wall and adjacent tissues, 1 of which also exhibited 2 metastatic lymph nodes in the central neck area. The cases reported illustrate the utility of enhancing one's clinical suspicion of carcinoma in patients bearing TDC, even when incidentally discovered. In particular, rapid growth of the cystic mass, and the presence of a mural nodule on US, especially with calcifications, must raise the physician's suspicion for a cancer arising in TDC. PMID- 12553554 TI - Pattern visual evoked potential (PVEP) evaluation in hypothyroidism. AB - Dysfunction of the central nervous system (CNS) is an important consequence of thyroid hormone deficiency. Evoked potentials like visual evoked potentials (VEP) provide a reliable and objective measure of function in related sensory system and tracts. In this study pattern-shift VEP (PVEP) recordings were performed on 48 newly diagnosed hypothyroid patients. Twenty-four had sub-clinical and 24 had overt hypothyroidism. None of the patients had clinical symptoms or signs referable to CNS dysfunction. Their mean age was 44+/-12 yr. The response to pattern stimulation on the normal control subjects was a triphasic response with a prominent positive wave (P100) with a peak latency of 84-105 (mean: 96+/-4) milliseconds (ms). In patients with hypothyroidism mean P100 latency was (mean: 97+/-6) ms and the difference between the 2 groups was not statistically significant. (p>0.05) Delays above the average latency +/-2.5 SD of the mean of the control subjects was defined as a criteria for an abnormality. According to defined criteria 6 (12.5%) patients demonstrated abnormal PVEP at least on one tested side. Previous studies conducted on small patient populations stated there is high percentage of VEP abnormalities in hypothyroid patients. However, this fact was not confirmed by our study. We believe abnormalities of PVEP will be more prominent in untreated patients in the advanced stage of the disease, or in patients who have a neurological involvement; such as apathy, impaired memory or cerebellar dysfunction. Consecutive studies, in a more clearly defined and selected patient population, are needed to confirm and settle this issue. PMID- 12553555 TI - Co-expression of interleukin-6 (IL-6) and interleukin-6 receptor (IL-6R) in thyroid nodules is associated with co-expression of CD30 ligand/CD30 receptor. AB - Data on the expression of interleukin 6 (IL-6)/interleukin 6 receptor (IL-6R) in thyroid nodules is scarce. Based on our recent data of CD30 ligand (CD30L)/CD30 receptor (CD30) in these nodules and on the knowledge that this signal stimulates IL-6 production in non-thyroid neoplasms, we wanted to evaluate the immunocytochemical expression of these 2 ligand/receptor systems in a large archival series of paraffin-embedded specimens. These specimens included 6 normal thyroids and 130 thyroid nodules. Co-expression of IL-6 and IL-6R in the epithelial (follicular) cells was observed solely in CD30L/CD30 positive nodules: 5/15 (33%) oncocytic adenomas; 6/30 (20%) follicular adenomas which belonged to 2 variants (4/4 microfollicular toxic and 2/2 hyalinizing trabecular); 9/30 (30%) papillary thyroid cancers (PTC), all belonging to the conventional variant. In PTC the proportion of tumor epithelial cells that were IL6 positive was inversely correlated with the pTNM staging (r=-0.549, p=0.01). All 15 follicular cancers (FTC), all 6 anaplastic cancers (ATC) were IL-6/lL-6R negative; 14/15 FTC and 5/6 ATC were CD30L/CD30 negative. In another oncocytic adenoma, another 4 conventional PTC and another 7 non-conventional PTC CD30L/CD30 expression was associated to expression of IL-6 only. IL-6 staining associated to absent expression of CD30L and CD30 was observed in 7 follicular adenomas (all belonging to variants different from toxic and hyalinizing trabecular), 2 oncocytic adenomas, 5 of the 30 colloid nodules and 2 normal thyroids. Of the 6 tumors arising from the parafollicular C cells (medullary thyroid cancer, MTC), all 3 that had metastasized were CD30L/CD30/IL-6 positive and IL-6R negative; only IL-6 expression was lost in both the local and distant metastases. This finding matched the loss of IL-6 expression in one PTC metastasis. All 3 non-metastasized MTC were IL-6/IL-6R negative, and 1/3 was CD30L positive/CD30 negative. We conclude that only in a subset of both benign and malignant thyroid nodules the IL-6/IL-6R signal could be induced by the CD30L/CD30. IL-6 expression is related with aggressiveness in both PTC and MTC. In the normal thyroid tissue, colloid nodules, and another subset of benign and malignant thyroid nodules, IL-6 expression is under control of signals other than CD30L/CD30. PMID- 12553556 TI - Reversible increase of serum activin A levels in women with Graves' disease. AB - The aim of this study was to analyze the serum levels of activin A in hyperthyroid patients with Graves' disease. Serum activin A and FSH levels were measured in a total of 93 females (64 regularly cycling and 29 post-menopausal). Of these, 20 were hyperthyroid patients with Graves disease, 33 were euthyroid goitrous patients (20 had autoimmune thyroiditis AT and 13 only had goiter) representing the internal control group and 40 were healthy subjects representing the external control group. Serum levels of activin A were higher in goitrous patients with AT than in control subjects (p=0.0388). Activin A levels were almost doubled in the cycling and in post-menopausal hyperthyroid women (0.91+/ 0.21 vs 0.43+/-0.07 microg/l; p<0.0001 and 0.92+/-0.22 vs 0.48+/-0.24 microg/l; p=0.0001, respectively). In 10 cycling hyperthyroid patients, studied even after methimazole treatment, that increase was substantially reversed, once euthyroidism was attained (p=0.002). These findings indicate that thyroid function and autoimmune processes significantly affect serum levels of activin A in patients with Graves' disease. PMID- 12553557 TI - Improvement of left ventricular hypertrophy and arrhythmias after lanreotide induced GH and IGF-I decrease in acromegaly. A prospective multi-center study. AB - We report the results of a prospective Italian multi-center study of the effects of lanreotide, a slow-release somatostatin analog, on left ventricular morphology and function and on the prevalence of ventricular arrhythmic events in 19 patients with active, newly diagnosed, uncomplicated acromegaly. Cardiac features were evaluated with Doppler-echocardiography and 24-h Holter ECG monitoring at baseline and after 6 months of lanreotide therapy. Fifteen patients (78.9%) had left ventricular hypertrophy. Lanreotide treatment significantly decreased the left ventricular mass (127.8+/-6.9 vs 140.7+/-7.1 g/m2, p<0.001) and left ventricular hypertrophy significantly disappeared in 6 of these patients. Treatment did not significantly affect systolic function, whereas it increased the Doppler-derived early-to-late mitral flow velocity, (E/A) ratio, of early-to late trans-mitral flow velocity (1.34+/-0.1 vs 1.09+/-0.06, p=0.001). Stroke volume was slightly but not significantly increased after treatment, whereas systolic BP was significantly higher (134+/-14 vs 129+/-13 mmHg, p<0.05). The 24 h mean heart rate was significantly reduced after treatment (66.5+/-11 vs 71.5+/ 20 beats/min, p<0.05). Supra-ventricular premature beats (>50/24 h) occurred in 16.6% of patients and were unaffected by treatment. Differently, ventricular premature beats (>50/24 h) occurred in 33.3% of patients before treatment vs 16.5%, after treatment. In conclusion, lanreotide reduced the left ventricular mass, and improved ventricular filling and ventricular arrhythmic profile. PMID- 12553558 TI - Heterogeneity of the thyroglobulin epitopes associated with circulating thyroid hormone autoantibodies in hashimoto's thyroiditis and non-autoimmune thyroid diseases. AB - We previously implicated TG leakage from fine-needle aspiration biopsy (FNAB) as responsible for circulating thyroid hormone autoantibodies (THAb). However, THAb were not always associated with TGAb. In the literature these negative findings have been interpreted against a role of TG as the antigen for THAb. To evaluate the TGAb status more fully and to gain information on TG epitopes involved in THAb development, we measured: 1) TGAb with an independent hemagglutination assay (HA), and 2) epitope specificity in a competitive ELISA using 2 monoclonal Abs (mAb) against TG: mAb 42C3 and mAb 134C2. MAb 42C3 recognizes a cross-reactive iodinated epitope, whereas 134C2 is specific for human TG. We tested 12 Hashimoto's thyroiditis (HT) and 35 non-HT patients sampled prior to, 1 and 3 months after FNAB. We found that, irrespective of thyroid disease or post-FNAB THAb status, certain patients previously classified as TGAb negative by IRMA tested TGAb positive by HA or by competition ELISA and vice versa. A post FNAB positive response to the 42C3 iodinated epitope in only one THAb IgM-T4+ve HT and a few THAb negative non-HT patients was observed. Furthermore, we observed that the 3 non-HT patients who expressed IgM-T3 THAb failed to bind either TG-mAb epitope. We conclude that a single TGAb assay is not sufficient to define the TGAb status, which can be achieved reliably only by using multiple TGAb assays. In addition, the TG-iodinated epitope recognized by 42C3 is not a major epitope in post-FNAB THAb, and the T3-epitope involved in THAb remains distinct from the mAb epitopes. In light of recent data in the literature, we further suggest that the responsible epitopes are more likely to be expressed in leaked TG fragments, rather than leaked intact TG. PMID- 12553559 TI - Glutathione peroxidase (GPX) activity in seminal plasma of healthy and infertile males. AB - Human spermatozoa are more dependent on glutathione peroxidase/glutathione reductase (GPX/GR) system, via reduced glutathione (GSH), to inactivate reactive oxygen metabolites (ROMs) such as hydrogen peroxide and organic hydroperoxides. To demonstrate whether there is a substantial difference in GPX activity between normal and pathological seminal plasma, we decided to evaluate the activity of this enzyme in healthy subjects and infertile males with normal hormonal patterns. Our results demonstrate that in healthy subjects the seminal plasma contains a GPX activity that is about 10 times greater than the GPX activity detected in the seminal plasma of infertile males. By using specific antibodies against plasmatic GPX (GPX3), we also demonstrate that enzymatic activity, detected in the seminal plasma of both healthy and infertile males is GPX3. Therefore, the evaluation of GPX activity in human seminal plasma could be a new useful marker of gonadal function in men. PMID- 12553560 TI - A mathematical model for pattern of change in beta-cell reserve and factors affecting residual reserve within the first 2 years of type 1 diabetes. AB - The aim of this study was to investigate the effects of age, duration of diabetes, sex and ICA (Islet cell cytoplasmic antibody) on beta-cell reserves and to develop a model within the first 2 years of Type 1 diabetes. Beta-cell reserve is evaluated as fasting (FCp) and 1 mg i.v. glucagon stimulated C-peptide (SCp) levels in 58 Type 1 diabetics and in 12 normoglycemic subjects. Patients were divided into 3 groups according to duration of diabetes: Group I (2.5+/-0.3 weeks), Group II (13.4+/-1.2 months) and Group III (24.2+/-1.8 months). FCp/SCp level in nmol/l (mean+/-SE) were as follows. Group I: 0.21+/-0.02/0.38+/-0.04, Group II: 0.15+/-0.01/0.27+/-0.02, Group III: 0.07+/-0.01/0.11+/-0.02, CONTROL GROUP: 0.42+/-0.09/1.29+/-0.13. The scatter plots of C-peptide levels vs time in all the diabetic patients fitted in to a 4th-order polynomial regression (R: 0.96 0.98). Age was strongly correlated with FCp (rs: 0.46, p<0.05) and ICA positivity affected Cp-levels negatively (p>0.05). In conclusion, as the duration of diabetes increases, response time to glucagon prolongs and amplitude of it shortens. Duration of diabetes of less than 2 weeks, feminity, puberty and ICA positivity affect beta-cell reserve negatively, conversely, masculinity, post puberty, older age and ICA negativity affect the reserve positively. The dynamics of C-peptide response to glucagon follow a mathematical model and Type 1 diabetes causes a decrease not only in the amplitude of the response but also in the duration of the response. PMID- 12553561 TI - Relationship between Y-chromosomal DNA haplotype and sperm count in Italy. AB - Forty Italian individuals with sperm counts in the range 20-130x10(6)/ml were typed with eleven Y-specific binary markers. Five Y haplogroups (1, 2, 3, 9 and 21) were present in the sample. In Italy, in contrast to Denmark, sperm counts were similar in the different haplogroups. PMID- 12553562 TI - Autoimmune thyroid disease accompanied by recurring episodes of painful thyroid ameliorated by thyroidectomy. AB - A 71-yr-old female initially presented with clinical and biochemical hyperthyroidism with high TSH binding inhibitory immunoglobulin (TBII) and anti thyroid peroxidase antibody (TPOAb) titers. Histological findings of the thyroid revealed hyperplasia with a focal germinal center, indicating Graves' disease and mild focal chronic thyroiditis. Four episodes of painful and tender thyroid occurred over the next 2 yr accompanied by acute inflammatory reactions. The first episode that developed while the patient was in a hyperthyroid state was soon followed by hypothyroidism associated with further increases in anti thyroglobulin antibody (TGAb) and TPOAb titers. The subsequent 3 episodes occurred during the hypothyroid state, when the TGAb titer progressively increased with each episode. We performed subtotal thyroidectomy to prevent further episodes. Specimens obtained at thyroidectomy showed that extreme fibrosis had replaced the thyroid parenchyma with collapsed follicles and moderate lymphocyte infiltration. No further episodes occurred after thyroidectomy, and during a 3-yr follow-up period, TBII and thyroid-stimulating antibody (TSAb) disappeared and TGAb and TPOAb titers decreased. This case report provides further evidence supporting the notion that thyroid epithelial destruction progresses during relatively short periods of recurrent painful thyroid and that thyroidectomy helps patients affected by this condition that are unresponsive to other treatment strategies. PMID- 12553563 TI - CTLA4 gene and autoimmune endocrinopathies: a new marker? PMID- 12553565 TI - Achondroplasic and GH-deficiency dwarfism. Diego Velasquez (1656). PMID- 12553566 TI - Outbreaks of Salmonella serotype enteritidis infection associated with eating shell eggs--United States, 1999-2001. AB - A Salmonella serotype Enteritidis (SE) epidemic emerged in the 1980s, when increasing numbers of infections were detected in the Northeastern and Mid Atlantic regions of the United States. In the early 1990s, while SE rates in the Northeast began to decline, the SE epidemic expanded to the Pacific region. Nationwide, the number of SE isolates reported to CDC peaked at 3.8 per 100,000 population in 1995. Although rates of culture-confirmed SE infection reported to CDC declined to 1.9 by 1999 (Figure 1), rates did not decline further through 2001, and outbreaks continue to occur. Investigations of outbreaks and sporadic cases have indicated repeatedly that, when a food vehicle is identified, the most common sources of SE infection are undercooked and raw shell eggs. This report describes two SE outbreaks associated with eating shell eggs and underscores the need to strengthen SE-control measures. PMID- 12553567 TI - Blood safety monitoring among persons with bleeding disorders--United States, May 1998-June 2002. AB - Since 1998, CDC has collaborated with approximately 140 federally funded hemophilia treatment centers (HTCs) in the United States and its territories through the Universal Data Collection (UDC) surveillance project to monitor blood product safety and detect new viral hepatitis and human immunodeficiency virus (HIV) infections. This report presents findings of investigations conducted during May 1998-June 2002 of 1,149 seroconversions for hepatitis viruses identified among persons with bleeding disorders who were enrolled voluntarily in UDC; 99% of hepatitis A virus (HAV) seroconversions and 90% of hepatitis B virus (HBV) seroconversions were attributed to vaccination programs against HAV or HBV. None of these cases was attributable to blood products received during this time, which indicates that the virally inactivated blood factor concentrates used to treat bleeding disorders are unlikely to transmit viral hepatitis. Regular monitoring of patients ensures the continued safety of blood and blood products. PMID- 12553564 TI - Disorders of ejaculation. AB - The physiology of ejaculation includes emission of sperm with the accessory gland fluid into the urethra, simultaneous closure of the urethral sphincters, and forceful ejaculation of semen through the urethra. Emission and closure of the bladder neck are primarily alpha-adrenergically mediated thoracolumbar sympathetic reflex events with supraspinal modulation. Ejaculation is a sacralspinal reflex mediated by the pudendal nerve. The most common ejaculation disorder is premature ejaculation, but there is little agreement regarding the definition of this disorder or its etiology, diagnosis, and treatment options. Premature ejaculation is in fact classically considered psychogenic in nature. However, recent data have demonstrated that prostatic inflammation/infection has been found with high frequency in premature ejaculation, suggesting a role of prostatic pathologies in the pathogenesis of some cases of failure of ejaculatory control. Rarer disorders are emission and ejaculation failure and urine contamination of semen. The new use of diagnostic procedures and the availability of pharmacological aids place this topic in the mainframe of medical sexology. PMID- 12553568 TI - Fusion to green fluorescent protein improves expression levels of Theileria parva sporozoite surface antigen p67 in insect cells. AB - East Coast fever (ECF) is a fatal disease of cattle caused by the protozoan parasite Theileria parva. The development of a subunit vaccine, based on the sporozoite-specific surface antigen p67, has been hampered by difficulties in achieving high-level expression of recombinant p67 in a near-authentic form. Therefore two sets of recombinant baculovirus vectors were constructed. The first set, encoding various regions of p67, produced low levels of the corresponding p67 domains in High Five cells, despite the presence of large amounts of p67 RNA. The second, consisting of p67 domains fused to the carboxy-terminus of GFP expressed significantly higher levels of p67 protein. The GFP:p67 fusion proteins were recognized by a sporozoite-neutralizing monoclonal antibody (TpM12) raised against native p67 whereas non-fused full length p67 expressed in insect cells was not recognized. GFP-tagging therefore, appeared to enhance the stability of p67 and to conserve its folding. The high-level expression of p67 domains in a more authentic form is an important step towards the development of an effective subunit vaccine against ECF. PMID- 12553569 TI - Concealing identity and mimicking hosts: a dual chemical strategy for a single social parasite? (Polistes atrimandibularis, Hymenoptera: Vespidae). AB - Polistes atrimandibularis is a species of obligate social wasp parasite, which introduces into nests of the host species P. biglumis, where it rears its own brood relying on the workers of the host species. Wasps of the host species discriminate between nestmates and non-nestmates by means of their epicuticular hydrocarbons. To overcome the host recognition systems, parasites have to modify their epicuticular recognition cues. Compared to individuals of the host species, parasites have 3-4 times less epicuticular hydrocarbons, although they share with their hosts the habitat and even the nests. We hypothesize that a dilution of the recognition cues in social parasites could have a facilitating role in entering host nests, in integrating within their host colony and in sneaking into secondary colonies to steal larvae and pupae. PMID- 12553570 TI - Carbohydrate inhibition of Biomphalaria glabrata embryonic (Bge) cell adhesion to primary sporocysts of Schistosoma mansoni. AB - Due to shared characteristics with snail haemocytes, the Biomphalaria glabrata embryonic (Bge) cell line has been used as a model in vitro system for the study of snail-trematode interactions. In this study, Bge cells were used to characterize the adherence of snail host cells to schistosome primary sporocysts, and to test the effect of carbohydrates as inhibitors of this behaviour. Bge cells bound to the surface of >90% of Schistosoma mansoni sporocysts and, based on a semi-quantitative cell adhesion scale of 1-4, exhibited a cell adhesion index (CAI) of 2.45. This cellular adhesion was significantly inhibited in the presence of selected carbohydrate-containing substances. Fucoidan was the most potent inhibitor, reducing Bge cell-binding prevalence to approximately 50% and the CAI to 1.6. Other inhibitory compounds included mannose-6-phosphate, heparin, dextran sulfate, and various forms of the polysaccharide carrageenan. Fluoresceinated-fucoidan was found to attach to Bge cells confirming their ability to associate with sugar moieties. These results were further supported by the specific binding of surface biotinylated Bge cell proteins to sporocyst tegumental glycoproteins ranging from 40 to 120 kDa. N-linked tegumental carbohydrates appeared to represent ligands for Bge cell proteins since N glycosidase treatment of blotted tegumental glycoproteins completely abolished biotinylated Bge protein binding. We hypothesize that the involvement of lectins as potential host cellular receptors in snail cell-sporocyst interactions, and suggest that negatively charged (mainly sulfated) carbohydrate moieties may represent the schistosome surface ligand(s). PMID- 12553571 TI - Functional visualization of the excretory system of adult Schistosoma mansoni by the fluorescent marker resorufin. AB - Excretion of metabolic wastes as well as xenobiotics is a major concern of all living organisms, and the Platyhelminthes including Schistosoma mansoni possess the protonephridial excretory system for their survival. Except for some ultra structural and biochemical information, little is known about the protonephridium of platyhelminths due to a lack of established techniques for exploring the excretory activity. This study describes a new technique to assess the excretory activity of S. mansoni by use of the fluorescent marker resorufin, which is a potential substrate of the drug efflux pump, P-glycoprotein. After simple diffusion into the schistosome body, fluorescent resorufin was concentrated in the excretory tubules by an energy-dependent mechanism and excreted via the nephridiopore. The present technique of labelling functionally the excretory system was applicable to adult worms, but not schistosomula or cercariae. A variety of modulators known to interfere with mammalian P-glycoprotein function perturbed resorufin excretion from male adult schistosomes, including cyclosporin A, Ro11-2933, verapamil, or nifedipine. This technique of labelling the excretory system with fluorescent resorufin has enabled us to study aspects of the physiological function, hitherto unknown, of the protonephridial system of S. mansoni. PMID- 12553572 TI - The influence of colostrum on early Schistosoma mattheei infections in calves. AB - The study investigated whether the susceptibility of calves to an early Schistosoma mattheei infection may be modified by intake of colostrum from infected cows. Twelve calves born to non-infected mothers were randomly divided into 2 groups of 6. The animals from group 1 were fed colostrum originating from a pool collected from non-infected cows, the calves from group 2 received colostrum from a pool collected from cows infected with S. mattheei. One month after birth all calves were infected by exposure to 1000 cercariae of a local strain of S. mattheei, and perfused 12 weeks later to determine the worm- and tissue egg counts. IgG(H+L), IgG1, IgG2 and IgA levels against soluble adult worm antigen preparation of S. bovis (SWAP bovis) were analysed in both colostrum pools and in the serum from the calves collected during the study before and after receiving colostrum, then on days 7, 30, 73 and 122. Faecal egg counts were determined from day 73 onwards. The IgG(H+L), IgG1 and IgA levels of the positive colostrum pool were higher than those of the negative pool. Calves of group 2 showed significantly higher levels of IgG(H+L) and IgG1 until day 73, to reach equal levels at necropsy. Calves of group 2 showed significant reductions of 42, 28 and 42% in total worm counts, female worm counts, and tissue egg counts, respectively, and a reduction of 25% in cumulative faecal egg counts. These findings indicate that there was a significant impact of colostrum on the parasitological and serological course of early S. mattheei infections. PMID- 12553573 TI - Detection of Onchocerca volvulus infection in low prevalence areas: a comparison of three diagnostic methods. AB - The standard assay for onchocerciasis diagnosis is microscopical detection of microfilariae in skin snips. Skin snipping is painful, requires appropriate sterilization of equipment, and may fail to diagnose light infections. Two alternatives are a polymerase chain reaction (PCR) test which detects parasite DNA in pieces or scrapings of skin and a test based on allergic reactions to topical application of diethylcarbamazine (DEC). We compared these 2 diagnostics with standard skin snip microscopy in 313 individuals from 2 villages in Guinea, with low prevalence after over 10 years of control by the Onchocerciasis Control Programme. Lower and upper bounds on sensitivities and specificities of these 3 tests were estimated. In addition, these parameters were estimated using 5 different statistical models. Where prevalence was low, PCR and the DEC patch test appeared to be more sensitive than skin snipping which has low sensitivity. As the DEC test is non-invasive, simple and cheap, it may provide a good alternative to skin snipping alone for surveillance in low prevalence areas. PMID- 12553574 TI - Vertical migration by the infective larvae of three species of parasitic nematodes: is the behaviour really a response to gravity? AB - Vertical migration by infective larvae (L3) of 3 species of nematodes was investigated. Upright truncated agarose cones were used to test upward migration, and comparable truncated cone-shaped agarose hollows were used to test downward migration. Flat agarose plates were control surfaces. When placed at the bases of upright cones, 74% of Ancylostoma caninum L3 migrated up, whereas only 16.5% migrated down the indented cones; this latter value was not significantly different from larval migration on flat plates. Strongyloides stercoralis L3 also migrated upward in significant numbers (80%). These larvae also failed to migrate downward under normal conditions. However, when the bottoms of the indented cones were 3-5 degrees C warmer than the tops, 75.5% of S. stercoralis L3 migrated down. In contrast, Haemonchus contortus L3 showed no tendency to crawl up or down cones, when compared with normal crawling behaviour on flat plates. Thus, L3 of A. caninum and S. stercoralis exhibited negative geotaxis, crawling against the pull of gravity, while H. contortus did not. The biology of these parasites may explain these behavioural differences. PMID- 12553575 TI - Neuronal uptake of pesticides disrupts chemosensory cells of nematodes. AB - Low doses of the acetylcholinesterase-inhibiting carbamate nematicides disrupt chemoreception in plant-parasitic nematodes. Fluorescein isothiocyanate (FITC)/dextran conjugates up to 12 kDa are taken up from the external medium by certain chemosensory neurons in Caenorhabditis elegans. Similar chemoreceptive neurons of the non-feeding infective stage of Heterodera glycines (soybean cyst nematode) fill with FITC and the nuclei of their cell bodies selectively stain with bisbenzimide. The widely used nematicide aldicarb disrupts the chemoreceptive response of H. glycines with 50% inhibition at very low concentrations (ca 1 pM), some 10(-6)-fold lower than required to affect locomotion. Similarly, the anthelmintic levamisole had this effect at 1 nM. Peptides selected as mimetics of aldicarb and levamisole also disrupt chemoreception in H. glycines and Globodera pallida at 10(-3)-fold or lower concentration than required to inhibit locomotion. We propose an uptake pathway for aldicarb, levamisole, peptide mimetics and other soluble molecules by retrograde transport along dendrites of chemoreceptive neurons to the cell bodies and synapses where they act. This may prove to be a general mechanism for the low dose effects of some nematicides and anthelmintics. PMID- 12553576 TI - A molecular phylogeny of nuclear and mitochondrial sequences in Hymenolepis nana (Cestoda) supports the existence of a cryptic species. AB - Since isolates of Hymenolepis nana infecting humans and rodents are morphologically indistinguishable, the only way they can be reliably identified is by comparing the parasite in each host using molecular tools. In the current study, isolates of H. nana from rodent and human hosts from a broad geographical range were sequenced at the ribosomal first internal transcribed spacer (ITS1), the mitochondrial cytochrome c oxidase subunit 1 (C01) gene and the nuclear paramyosin gene loci. Twenty-three isolates of H. nana were sequenced at the ITS1 locus and this confirmed the existence of spacers which, although similar in length (approximately 646 bp), differed in their primary sequences which led to the separation of the isolates into 2 clusters when analysed phylogenetically. This sequence variation was not, however, related to the host of origin of the isolate, thus was not a marker of genetic distinction between H. nana from rodents and humans. Sequencing of a 444 bp fragment of the mitochondrial cytochrome c oxidase 1 gene (C01) in 9 isolates of H. nana from rodents and 6 from humans identified a phylogenetically supported genetic divergence of approximately 5% between some mouse and human isolates. This suggests that H. nana is a species complex, or 'cryptic' species (=morphologically identical yet genetically distinct). A small segment of the nuclear gene, paramyosin, (625 bp or 840 bp) was sequenced in 4 mouse and 3 human isolates of H. nana. However, this gene did not provide the level of heterogeneity required to distinguish between isolates from rodent and human hosts. From the results obtained from faster evolving genes, and the epidemiological evidence, we believe that the life cycle of H. nana that exists in the north-west of Western Australia is likely to involve mainly 'human to human' transmission. PMID- 12553577 TI - On alert for bluetongue. PMID- 12553578 TI - Hip dysplasia in labrador retrievers: the effects of age at scoring. AB - Selective breeding policies for preventing or controlling hip dysplasia require accurate estimates of parameters in offspring/parental relationships and estimates of heritability. Recent literature includes some major studies of pedigree breeds of dog, using data derived from the hip dysplasia screening scheme set up by the British Veterinary Association. These publications have not taken into account the age of the animals when they were screened. This study analyses the data from 29,213 labrador retrievers whose ages were known when they screened. The mean hip score of the dogs was positively and significantly correlated with their age. If this relationship with age is ignored, various offspring/parental relationships and the estimates of heritability are likely to be distorted. PMID- 12553579 TI - Field study of the efficacy of three types of nose net for the treatment of headshaking in horses. AB - Thirty-six owners of seasonally headshaking horses took part in a trial to compare the effectiveness of three types of nose net, a traditional cylindrical net (full net) and two forms of larger mesh nets which cover only the nostrils and dorsorostral muzzle (half nets). Baseline data relating to the overall severity of the problem and 18 specific behaviours describing the nature of the problem were recorded on a check sheet by the owners. A within-subjects repeated measures design experiment, with each net used for a week before reassessment, was then used to assess the effect of the nets on the headshaking problem. Approximately 75 per cent of owners reported some overall improvement with each net; around 60 per cent recorded a 50 per cent or greater improvement and 30 per cent a 70 per cent or greater improvement. The nets significantly reduced the overall headshaking score and the following specific behaviours: up-and-down headshaking, nose flipping, acting as if a bee had flown up the nose, shaking at exercise, shaking when excited, shaking in bright sunlight or in windy conditions (P < 0.0001), striking at the face, shaking at night, rubbing the nose when moving, rubbing the nose on objects, sneezing, shaking in the rain and shaking indoors (P < 0.05). There was no evidence of a significant effect on side-to-side headshaking, shaking at rest or rubbing the nose when stationary, but the effect on snorting was uncertain. There were few significant differences between the nets, but the half nets were reported to be significantly better at controlling 'bee up the nose' behaviour. Horses more than 10 years old were reportedly less likely to show a 50 per cent or greater improvement in 'nose flipping' and 'headshaking at exercise. PMID- 12553580 TI - Sevoflurane anaesthesia in chickens during spontaneous and controlled ventilation. AB - A crossover study design was used to investigate the dose-related effects of sevoflurane at end-tidal concentrations of 2.2 to 4.4 per cent on the respiratory rate, blood gases, heart rate, arterial blood pressure and ocular signs of chickens during spontaneous and controlled ventilation. The mean (sd) carbon dioxide partial pressure (PaCO2) increased as the concentration of sevoflurane increased, and was 86 (29) mmHg at an end-tidal concentration of 4.4 per cent during spontaneous ventilation, but was maintained between 29 and 42 mmHg during controlled ventilation. The heart rate increased as the concentration of sevoflurane increased during spontaneous ventilation, but did not change during controlled ventilation. Sevoflurane decreased arterial blood pressure during both spontaneous and controlled ventilation, but a dose-dependent decrease in arterial blood pressure was observed only during controlled ventilation. The mean arterial blood pressure at an end-tidal concentration of 4.4 per cent was significantly higher during spontaneous ventilation than during controlled ventilation. Controlled ventilation prevented the increases in PaCO2 and heart rate that were observed during spontaneous ventilation. The decrease in arterial blood pressure during spontaneous ventilation was less than that during controlled ventilation, possibly owing to the effects of hypercapnia. PMID- 12553581 TI - Diagnosis of canine parvovirus by rapid immunomigration on a membrane. AB - Rapid immunomigration on a membrane was applied to the diagnosis of canine parvovirus (CPV) in 128 samples of faeces containing four strains of parvovirus (two CPV-2a strains, including one vaccine strain, and two CPV-2b strains). The results were compared with the results of haemagglutination and ELISA sandwich techniques. The new test was quick and easy to use, and made it possible to identify both the CPV-2a and CPV-2b strains. Its detection thresholds per gram of faeces corresponded to specific haemagglutination titres of between 320 and 640 and a virus titre of between 10(4) and 10(5) CCID50 (dose required to infect 50 per cent of cell cultures). PMID- 12553582 TI - Survey of sheep nematode parasite control methods in south-east Scotland. PMID- 12553583 TI - Efficacy of cyclosporine for treatment induction of canine pemphigus foliaceus. PMID- 12553584 TI - Mammary botryomycosis and mastectomy in an African elephant (Loxodonta africana). PMID- 12553585 TI - Meningitis associated with Salmonella Newport in a neonatal alpaca (Lama pacos) in the United Kingdom. PMID- 12553586 TI - Bovine TB: studies on the risk of herd breakdown. PMID- 12553587 TI - Eradication of scrapie in a Massese sheep flock by PrP allele selection. PMID- 12553589 TI - Contextualized behavioral support in early intervention for children with autism and their families. AB - Parent education programs have become an effective mode of treatment delivery for teaching families effective behavioral strategies to manage challenging behavior in young children with autism. Functional assessment and functional communication training (FCT) are empirically validated procedures that have recently been introduced into parent education programming to help resolve challenging behaviors. The success of these procedures, however, is contingent on family members' ability to integrate them into the specific contexts in which challenging behaviors occur. Consequently, the application of these procedures in home settings necessitates consideration of the family context in the assessment and treatment planning process. A study is presented that investigated the use of information on family context (i.e., caregiving demands, family support, patterns of social interaction) to direct the assessment and intervention planning process. More specifically, information on family context was used to individualize behavioral support plans designed to support family use of functional communication training within important family routines. Through parent-investigator collaboration we individualized the manner in which functional communication training procedures were taught and implemented so they were contextually relevant. Utilizing a multiple baseline design, the challenging behaviors and functional communication of three children with autism were monitored across baseline, intervention (i.e., FCT, and contextulized FCT), and follow-up phases. Multiple routines for each participant were selected and monitored across all phases to evaluate changes in the dependent measures within training and generalization routines. A self-report questionnaire was administered intermittently to parents to determine if consideration of family context improved the "goodness of fit" of the functional communication training treatment packages across FCT and contextualized FCT intervention phases. Results from the study indicate that consideration of family context in the assessment and intervention planning process does not jeopardize and may contribute to the stability and durability of reductions in challenging behavior achieved with functional assessment and functional communication training procedures. PMID- 12553588 TI - Eye movement abnormality suggestive of a spatial working memory deficit is present in parents of autistic probands. AB - Autistic probands exhibit impaired spatial accuracy and impaired response suppression errors during a delayed oculomotor response task. Family members of autistic probands, and thus the possible familial nature of these deficits, have not been assessed. Eleven parents of autistic probands and 17 adults from unaffected families, ages 25-50 years, completed oculomotor delayed-response tasks. Parents of autistic probands demonstrated poorer spatial accuracy than the comparison group (p = .002), with no significant differences between groups on percentage of premature saccades or latency of remembered saccades. Spatial working memory deficits, as measured by the delayed oculomotor response task, appear to be familial in families with an autistic proband. These deficits deserve further evaluation as a potential endophenotypic marker for genetic risk for autism. PMID- 12553590 TI - Decreasing disruptive behaviors of children with autism using social stories. AB - Few studies have been conducted in the area of social stories, and the limitations of these studies (i.e., improper story construction and methodological confounds) raise questions about effectiveness of social story interventions. This study examined the effectiveness of properly constructed social stories that have been introduced into the natural environment to target the disruptive behavior of three children with autism. A multiple baseline design across participants was employed, and a decrease in disruptive behavior was evidenced when the intervention was implemented for all participants. Based on the results of the present research, future areas of investigation outlining the limitations and potential benefits of social stories were discussed. PMID- 12553591 TI - Lack of benefit of intravenous synthetic human secretin in the treatment of autism. AB - The objective of this study was to determine if an intravenous infusion of synthetic human secretin improves language and behavioral symptoms in children with autism. Forty-two children with the diagnosis of autism were randomized to one of two groups in this double-blind cross-over trial. One group received 2 IU/kg of intravenous synthetic human secretin at the first visit, followed by an equal volume of intravenous saline placebo at week 6. The other group received treatments in the reverse order. All children were evaluated at weeks 1, 3, 6, 9, and 12 with standardized assessments of language, behavior, and autism symptomatology. There were no significant differences in the mean scores on any measure of language, behavior, or autism symptom severity after treatment with secretin compared to treatment with placebo. The results of this study do not support secretin as a treatment for autism. PMID- 12553592 TI - Sleep patterns of children with pervasive developmental disorders. AB - Data on sleep behavior were gathered on 100 children with pervasive developmental disorders (PDD), ages 2-11 years, using sleep diaries, the Children's Sleep Habits Questionnaire (CSHQ), and the Parenting Events Questionnaire. Two time periods were sampled to assess short-term stability of sleep-wake patterns. Before data collection, slightly more than half of the parents, when queried, reported a sleep problem in their child. Subsequent diary and CSHQ reports confirmed more fragmented sleep in those children who were described by their parents as having a sleep problem compared to those without a designated problem. Interestingly, regardless of parental perception of problematic sleep, all children with PDD exhibited longer sleep onset times and greater fragmentation of sleep than that reported for age-matched community norms. The results demonstrate that sleep problems identified by the parent, as well as fragmentation of sleep patterns obtained from sleep diary and CSHQ data, exist in a significant proportion of children with PDD. PMID- 12553593 TI - Semantic fields in low-functioning autism. AB - Restricted semantic fields and resultant stimulus overselectivity are often thought to be typical of low-functioning autism, as is a strong visual processing preference. However, these conclusions may in part be an artifact of testing methodology. A 12-year-old, low-functioning and nonverbal autistic boy was tested on an auditory word-to-picture selection task. The picture foils were chosen to have visual features, semantic features, both, or neither in common with the correct answer. Errors were made more often to semantically than to visually related items, and he showed generalization to items that had not been explicitly trained. This is taken as evidence that his semantic fields are broader than otherwise apparent, and that he was capable of expanding his semantic representations independently of specific training. PMID- 12553594 TI - Understanding embarrassment among those with autism: breaking down the complex emotion of embarrassment among those with autism. AB - Scenarios manipulating various factors within the emotion of embarrassment, such as whether or not an audience was present when an embarrassing act was committed, the type of audience present, empathic embarrassment, etc., were presented to high-functioning participants with autism and comparison groups of those with learning difficulties and typically developing participants matched for verbal and nonverbal mental age. Participants were required to rate the level of embarrassment of the protagonist and justify their responses. It was predicted that those with autism would differ significantly from the comparison groups in their ratings and also their ability to provide justifications. The results showed those with autism to have difficulty with such concepts as empathic embarrassment but showed a surprisingly good understanding of other variables manipulated such as the presence of an audience. PMID- 12553595 TI - Utility of the Gilliam Autism Rating Scale in research and clinical populations. AB - The Gilliam Autism Rating Scale (GARS) was developed as a relatively easy, inexpensive aid in the surveillance and diagnosis of autism. This study examined the validity of the GARS when used with a sample of 119 children with strict DSM IV diagnoses of autism, ascertained from both clinical and research settings. The GARS consistently underestimated the likelihood that autistic children in this sample would be classified as having autism. The sample mean for the Autism Quotient, a hypothesized index of the likelihood of having autism, was 90.10, significantly below the reference mean of 100. Diagnostic classification according to criteria specified by the GARS resulted in a sensitivity of only .48. Limitations of rating scales in general and of the GARS specifically are discussed. It is recommended that clinicians and researchers using or considering using the GARS for autism diagnosis or ratings of autism severity recognize the need for further research regarding its use. PMID- 12553596 TI - Assessing emotional and behavioral problems in children with intellectual disability: revisiting the factor structure of the developmental behavior checklist. AB - The objective of the reported study was to reassess the factor structure of the Developmental Behaviour Checklist (DBC) in a large cross-cultural sample representing all levels of intellectual disability. Parent and teacher DBC ratings on a combined sample of 1536 Dutch and Australian children and adolescents (ages 3-22) with mild to profound intellectual disability were used. Principal components analyses produced five subscales: Disruptive/Antisocial, Self-Absorbed. Communication Disturbance, Anxiety, and Social Relating, explaining 43.7% of the total variance. Internal consistencies of these subscales ranged from .66 to .91. The revised factor structure of the DBC appears to be an improved and useful tool for assessing emotional and behavioral problems in children with intellectual disabilities. PMID- 12553597 TI - Review of the Asperger Syndrome Diagnostic Scale. AB - The Asperger's Syndrome Diagnostic Scale (2001) was reviewed to determine it's reliability, validity, and clinical utility in the diagnostic process for pervasive developmental disorder, specifically Asperger's syndrome. Concerns were raised about validity of the instrument, population upon which the instrument was normed, capability of the instrument to provide accurate differential diagnoses, and properties of the scale. The measure was determined to hold promise as a research tool, but there appears to be little evidence that it can distinguish among the various types of pervasive developmental disorder or diagnosis Asperger syndrome specifically. PMID- 12553598 TI - Brain anatomy in autism. PMID- 12553600 TI - Effect of beam size on the expected benefit of customized laser refractive surgery. AB - PURPOSE: Customized laser surgery attempts to correct higher order aberrations, as well as defocus and astigmatism. The success of such a procedure depends on using a laser beam that is small enough to produce fine ablation profiles needed to correct higher order aberrations. METHODS: Wave aberrations were obtained from a population of 109 normal eyes and 4 keratoconic eyes using a Shack-Hartmann wavefront sensor. We considered a theoretical customized ablation in each eye, performed with beams of 0.5, 1.0, 1.5, and 2.0 mm in diameter. We then calculated the residual aberrations remaining in the eye for the different beam sizes. Retinal image quality was estimated by means of the modulation transfer function (MTF), computed from the residual aberrations. Fourier analysis was used to study spatial filtering of each beam size. RESULTS: The laser beam acts like a spatial filter, smoothing the finest features in the ablation profile. The quality of the correction declines steadily when the beam size increases. A beam of 2 mm is capable of correcting defocus and astigmatism. Beam diameters of 1 mm or less may effectively correct aberrations up to fifth order. CONCLUSION: Large diameter laser beams decrease the ability to correct higher order aberrations. A top-hat laser beam of 1 mm (Gaussian with FWHM of 0.76 mm) is small enough to produce a customized ablation for typical human eyes. PMID- 12553599 TI - Femtosecond laser flap creation for laser in situ keratomileusis: six-month follow-up of initial U.S. clinical series. AB - PURPOSE: To evaluate clinical performance of a femtosecond laser for flap creation in laser in situ keratomileusis (LASIK). METHODS: A prospective, consecutive series of 208 eyes (122 patients) undergoing LASIK between June 2000 and November 2000 using a femtosecond laser for creation of the corneal flap were evaluated for the incidence of complications. A subset of 114 patients who underwent myopic procedures and who were followed for 6 months was analyzed using standard outcome measures. RESULTS: In 4 of 208 eyes (1.9%), suction was lost during the procedure, causing the flap resection to be interrupted. However, in all four of these eyes the procedure was successfully performed 5 to 45 minutes after re-applanation of the eye. No postoperative complications or adverse events were observed in any treated eye. In the subgroup of 96 eyes undergoing plano correction and followed for 6 months (preoperative spherical equivalent refraction range -0.63 to -12.40 D), 98% (94 eyes) achieved uncorrected visual acuity of 20/40 or better; 94% (90 eyes) achieved 20/30 or better, 79% (76 eyes) achieved 20/25; and 55% (53 eyes) achieved 20/20 or better--all without benefit of retreatment. CONCLUSION: Femtosecond laser corneal flap creation is a safe and effective alternative to traditional mechanical microkeratomes. Standard LASIK nomograms appeared to apply equally well to the all-laser LASIK procedures. The potential for improved flap safety, reproducibility, flexibility, as well as for additional corneal applications are now being explored. PMID- 12553601 TI - Corneal topographic and pachymetric screening of keratorefractive patients. AB - PURPOSE: To review the incidence of corneal abnormalities detected in the preoperative examination, using videokeratography and pachymetry that excluded patients from laser in situ keratomileusis (LASIK) or photorefractive keratectomy (PRK). METHODS: We conducted a chart review of consecutive patients who had a preoperative examination and were excluded from refractive surgery. RESULTS: Eighteen patients from 1,392 refractive candidates (1.3%) were designated as poor candidates for LASIK or PRK based on corneal topography and/or pachymetry. Thirteen patients (0.9%) were identified as having keratoconus suspect, keratoconus, or pellucid marginal degeneration. Five patients (0.4%) were excluded due to thin corneas (<490 microm) despite normal corneal topography. CONCLUSIONS: Corneal topography and pachymetry are indispensable tools in the preoperative screening of refractive surgery candidates. PMID- 12553602 TI - Diffuse lamellar keratitis and flap margin epithelial healing after laser in situ keratomileusis. AB - PURPOSE: To evaluate whether wound healing of the corneal epithelium at the flap margin influences development of diffuse lamellar keratitis (DLK) after laser in situ keratomileusis (LASIK). METHODS: We examined prospectively 1928 eyes of 994 patients after LASIK. Condition of wound healing at the flap margin 1 day after LASIK was graded as gutter score from 0 to 3, according to the extent of fluorescein staining. Existence of DLK, gutter score, and the location of DLK and gutter were evaluated. RESULTS: Sixty-eight eyes (3.5%) developed DLK. The average gutter score was significantly higher in patients with DLK compared to those without DLK (P<.001). The location of DLK was identical to the location of gutter in 44 eyes (77.2%). CONCLUSION: Disruption of integrity of the epithelial barrier function at the flap margin was associcated with diffuse lamellar keratitis after laser in situ keratomileusis. PMID- 12553603 TI - Effect of hyperopic photorefractive keratectomy on corneal sensitivity: a longitudinal study. AB - PURPOSE: To investigate corneal sensitivity after photorefractive keratectomy (PRK) for low hyperopia, as measured with a non-invasive stimulus. METHODS: Two experimental groups were recruited: a control group of 17 patients (mean age 61.65 years) who underwent no treatment, and a PRK group of 11 patients (mean age 58.64 years) who underwent one of three attempted hyperopic corrections: +2.00 D (two patients), +3.00 D (four patients), +4.00 D (five patients). Corneal sensitivity was assessed centrally and peripherally, at temporal, medial, and inferior locations, approximately 1 mm from the limbus, using the Non-Contact Corneal Aesthesiometer (NCCA). Measurements were taken at each location for the control group and at preoperative, and postoperative weeks 1 and 2, 1, 3, and 6 months for the PRK group. RESULTS: Comparison of control and PRK groups (preoperative sensation threshold) (t-test): central P=.715, temporal P=.719, medial P=.943, inferior P= .920. Comparison of longitudinal changes in PRK group (one-way ANOVA): central P=.612, temporal P=.997, medial P=.981, inferior P=.993. CONCLUSIONS: Using the Non-Contact Corneal Aesthesiometer, no significant difference was found between the control and PRK groups for preoperative sensation thresholds, and no significant change in corneal sensitivity was found between any of the test time periods at any of the four corneal test locations for the PRK group. PMID- 12553604 TI - Phototherapeutic keratectomy with mitomycin C for corneal haze following photorefractive keratectomy for myopia. AB - PURPOSE: To evaluate the safety and efficacy of phototherapeutic keratectomy (PTK) with single application of mitomycin C for patients with severe corneal haze following photorefractive keratectomy (PRK) for high myopia. METHODS: Eight eyes of seven patients were treated with PTK and intraoperative topical application of mitomycin C (0.02%) for severe corneal haze (grade 3) following PRK for myopia. RESULTS: All patients' visual performance improved significantly. Mean preoperative visual acuity (20/200 for both UCVA and BSCVA) improved significantly to 20/33 (0.6) and 20/30 (0.7) for UCVA and BSCVA, respectively. Six eyes (85.7%) had improved UCVA to 20/40 or better and gained five or more lines of UCVA. The corneal haze score decreased from grade 3 initially (for all eyes prior to PTK and mitomycin C) to a final mean haze score of 0.3 (range 0 to 0.5). Mean final spherical equivalent refraction achieved was -1.30 +/- 1.60 D (range -3.75 to +1.25 D). One eye gained only three lines of visual acuity due to regression and residual haze. No adverse effects related to the use of mitomycin C were recorded. CONCLUSION: PTK with a single intraoperative application of mitomycin C was safe and effective in reducing corneal haze and improving visual acuity in patients with severe corneal haze following PRK. PMID- 12553605 TI - Results of laser in situ keratomileusis for myopia of -10 to -19 diopters with a Technolas 217 laser. AB - PURPOSE: To evaluate the effectiveness, predictability, and safety of laser in situ keratomileusis (LASIK) for correcting myopia greater than -10.00 D. METHODS: Sixty-five eyes of 37 patients with myopia greater than -10.00 D underwent LASIK. Patients were evaluated on day 1, 1 week, 1, 3, and 6 months after surgery. Parameters evaluated were uncorrected visual acuity (UCVA) and best spectacle corrected visual acuity (BSCVA), residual refractive error, regression of correction, and presence of any complication. RESULTS: Mean preoperative BSCVA was 0.745 +/- 0.234, which improved to 0.8070 +/- 0.237 postoperatively. The average preoperative UCVA was 0.022 +/- 0.02; postoperative UCVA at 6 months was 0.536 +/- 0.255. UCVA of 20/40 or better was achieved in 58% (38 eyes) and 20/20 or better in 26% (17 eyes). The average refractive error before LASIK was -12.64 +/- 2.16 D (range -10.00 to -19.00 D). Mean residual refractive error 1 week following LASIK was -0.63 +/- 1.36 D, which regressed to a mean -1.78 +/- 2.08 D at the end of 6 months. Nineteen eyes (29%) were within +/-0.50 D of intended refractive correction. CONCLUSION: LASIK was partially effective in the correction of high myopia. An initial overcorrection may be programmed to offset the effect of refractive regression. PMID- 12553606 TI - The ascent of Mount Everest following laser in situ keratomileusis. AB - PURPOSE: To report the visual experiences of climbers with prior laser in situ keratomileusis (LASIK) for myopia at extreme altitudes, including the summit of Mount Everest. METHODS: We measured the visual acuity of 12 LASIK eyes of 6 Mount Everest climbers at base camp (17,600 ft). Results are reported on their subjective visual experiences, as all climbers ascended above 26,000 feet and four reached the 29,035-foot summit. RESULTS: Five of the six climbers reported no visual changes up to 26,400 feet. Three climbers noted no problems and perfect vision with their LASIK eyes on the summit of Mount Everest. One reported mild blurring with ascent above altitudes of 16,000 feet that improved with descent, or a prolonged stay at altitude. Two climbers reported blurred vision at 27,000 and 28,500 feet, respectively, which improved with descent. CONCLUSION: Laser in situ keratomileusis may be a good choice for patients involved in high altitude activities. Patients achieving extreme altitudes of 26,000 feet and above should be aware of possible fluctuation of vision. PMID- 12553607 TI - Anterior lamellar keratoplasty with a microkeratome: a method for managing complications after refractive surgery. AB - PURPOSE: To demonstrate a technique of anterior lamellar keratoplasty with standardized and automated preparation of surface-parallel cuts in both donor and recipient appropriate for addressing several problems after laser in situ keratomileusis (LASIK) and photorefractive keratectomy (PRK). METHODS: We report a noncomparative series of ten eyes with complications after LASIK and PRK. Lamellar cuts were performed in donor and recipient eyes by means of an automated microkeratome. Lamellar grafts were fixed by only four single sutures. In two eyes, a re-lift LASIK was performed after 6 months. RESULTS: Surgery was uneventful and visual acuity was improved in all eyes. Residual irregular astigmatism and refractive error were corrected in two eyes by means of excimer laser computer-assisted ablation and resulted in a further improvement of uncorrected and best spectacle-corrected visual acuity. CONCLUSIONS: Anterior lamellar keratoplasty with a microkeratome can be used for the management of certain complications of PRK and LASIK. PMID- 12553609 TI - Equation for corneal asphericity after corneal refractive surgery. AB - PURPOSE: To determine the exact theoretical asphericity after refractive surgery when the parabolic approximation of the Munnerlyn formula is utilized. METHOD: From a least-square analysis and considering the conicoid model for anterior cornea, we calculated the final asphericity (p'-factor) from the preoperative asphericity and the radii before and after surgery. RESULTS: The final p'-factor is given by p'=p(R'3/R3); R' is the final radius of curvature; R is the preoperative radius; and p is the preoperative p'-factor. CONCLUSIONS: This equation for postoperative asphericity may be useful in studying the influence on asphericity of different surgical variables. It allows comparisons of real asphericity with the asphericity that should theoretically result. It also explains the increased spherical aberration after refractive surgery and the deterioration of retinal image quality that some authors have found experimentally. PMID- 12553608 TI - Photorefractive keratectomy with and without smoothing: a bilateral study. AB - PURPOSE: To assess whether a smoother stromal surface and a faster epithelium regeneration after excimer laser photorefractive keratectomy (PRK) may lead to better visual results. METHODS: Ten patients had unilateral PRK and contralateral PRK plus smoothing. The operative outcome was checked by means of digital standardized photographs taken at 0, 20, 40, and 60 hours after surgery. Complete clinical examinations were performed before and 1 and 3 months after surgery. RESULTS: Between 20 and 40 hours after surgery, the average velocity of radial epithelial migration in eyes in which smoothing was performed was 0.083 +/- 0.008 mm/h. This was faster than that (0.064 +/- 0.007 mm/h [P<.01]) observed in the eyes without smoothing. The corneal regularity index at 1 and 3 months was worse in the PRK alone group compared to the PRK plus smoothing group (P=.1). At 3 months, better visual outcome and less haze were observed in the PRK plus smoothing group. CONCLUSION: When using a flying spot laser, PRK plus smoothing improved the visual results and diminished regression and haze at 3 months, but gave rise to a slight hyperopic shift. PMID- 12553610 TI - Diffuse lamellar keratitis 6 months after uneventful laser in situ keratomileusis. AB - PURPOSE: Diffuse lamellar keratitis after laser in situ keratomileusis (LASIK) typically occurs between 1 and 7 days after the procedure, and its etiologic factor(s) remain unknown. METHODS: We describe a case of diffuse lamellar keratitis 6 months after uneventful LASIK in a 25-year-old woman. RESULTS: Slit lamp microscopy showed a diffuse infiltrate confined to the interface, extending to the visual axis, with no other relevant findings. Late on-set diffuse lamellar keratitis was our provisional diagnosis and treatment with topical corticosteroids was instituted, with rapid response and improvement of the clinical signs and symptoms. CONCLUSIONS: This case supports the theory that a previously inert inciting agent could cause a delayed toxic or inflammatory response of the cornea several months after surgery. PMID- 12553611 TI - Hansatome excursion in narrow palpebral apertures without lid speculum. PMID- 12553612 TI - Cataract following posterior chamber phakic intraocular lens. PMID- 12553613 TI - The "metal-free" myth. PMID- 12553614 TI - Guided tissue regeneration-based root coverage utilizing collagen membranes: technique and case reports. AB - Gingival recession defects have traditionally been treated with various grafting procedures. Recently, guided tissue regeneration with collagen membranes has shown promising results. This article reviews the rationale, indications, contraindications, and clinical methods for the use of bioabsorbable collagen membrane barriers. Several properties make collagen membranes attractive candidates for use as barriers in guided tissue regeneration-based root coverage procedures. These include the inhibition of epithelial migration and promotion of new connective tissue attachment; the ability to aggregate platelets, thereby facilitating wound stabilization and maturation; the promotion of cellular migration and wound closure; the elimination of the need for reentry surgery; and the ability to augment tissue thickness. Cases are presented to illustrate the surgical principles and techniques. PMID- 12553616 TI - Preliminary evaluation of salivary pellicle on nickel-chromium alloy in vivo. AB - OBJECTIVE: The composition of the salivary interface (pellicle) between dental restorations and oral mucosa may be critical to the biocompatibility of the restoration. The purpose of this study was to examine the molecular composition of the salivary pellicle on nickel-chromium alloy in vivo. METHOD AND MATERIALS: The molecular components of nickel-chromium pellicle was examined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analyses. RESULTS: Only limited numbers of salivary proteins were found to participate in the formation of nickel-chromium pellicle in vivo. Salivary amylase and secretory immunoglobulin A were among the proteins identified in the pellicle. CONCLUSION: In vivo, nickel-chromium pellicle consists of selectively adsorbed salivary proteins. Because both salivary amylase and secretory immunoglobulin A are antimicrobial proteins, it is possible that they play a role in modulating the microbial flora on the nickel-chromium prosthesis. PMID- 12553615 TI - Porcelain-veneered computer-generated partial crowns. AB - OBJECTIVE: It would be advantageous to be able to use computer-aided design and manufacturing to fabricate a restoration that can be layered with a conventional porcelain veneer in the occlusal region, thus optimizing esthetics, function, and strength. This case study reports the laboratory technique and the clinical performance of 38 partial crowns fabricated with computer technology and veneered with porcelain. METHOD AND MATERIALS: Twenty-one mandibular and 17 maxillary molars in 27 patients were prepared for partial crowns. The occlusal surfaces were lowered (1.5 to 2.0 mm), deep shoulders (1.5 mm) were prepared around the functional cusps, and 1.0-mm-deep shoulders were prepared in the proximal gingival regions. The nonfunctional cusps were prepared with an occlusal shoulder at approximately a right angle with the axial surfaces of the seat. In the computer-aided design procedure, the occlusal table was reduced to 1.4 mm above the preparation surface. The marginal ridge points, the marginal ridge line, the equator line, and the fissure line heights were adjusted accordingly. RESULTS: The lowest occlusal table thickness was 1.1 mm in six partial crowns, 1.2 mm in 26 partial crowns, and 1.3 mm in six partial crowns. The lowest occlusal table thickness of the porcelain veneers varied between 0.4 and 0.6 mm. The total occlusal table thickness thus was 1.5 mm or more. Clinically, no fractures occurred during an observation period varying between 1 and 4 years after placement. CONCLUSION: Computer-aided design and manufacturing technology is also convenient for partial crown preparation design with shoulder finish lines. PMID- 12553617 TI - A review of esthetic pontic design options. AB - Advances in the field of restorative materials allow a lost tooth to be replaced by artificial tooth structure that is virtually indiscernible from the original. However, in fixed partial dentures the standards for the pontic area and the adjacent soft tissue in the maxillary anterior region have increased in particular. The pontic design in this region is primarily influenced by esthetic and phonetic considerations. Local defects of the alveolar ridge often complicate restorative measures. Treatment methods proposed to solve this problem involve modification of the pontic design and pretreatment of the recipient site for the pontic. This article reviews the different clinical and technical options that are available for designing esthetic and functional pontics for the anterior region. PMID- 12553618 TI - An overview of esthetics with removable partial dentures. AB - A removable partial denture should restore function, phonetics, and esthetics. To assist the clinician in providing a restoration that is as esthetic as possible, despite the limitations of the conventional removable partial denture, this review collates the recommendations scattered among the literature. The review describes the steps involved in providing an esthetic restoration, including diagnosis and treatment planning; surveying; mouth preparation; framework and clasp alloys; clasps; rests; major connectors; minor connectors; prosthetic teeth; and denture base resin and flanges. The esthetic challenges of the Kennedy Class IV removable partial denture are also discussed. PMID- 12553619 TI - Holdaway measurement norms in Turkish adults. AB - OBJECTIVE: The aims of this study were to establish the cephalometric norms for Turkish adults by means of the Holdaway analysis and to investigate the cephalometric differences between Turkish women and men. METHOD AND MATERIALS: The study materials consisted of 100 cephalometric head films taken at the natural head position. The 50 men and 50 women who volunteered for the study had to be older than 18 years and had to have parents of Turkish origin, acceptable facial structure, ideal dental occlusion, no visual or hearing disorders, and all the teeth present in their mouths except the third molar teeth. The selected subjects had not undergone orthodontic treatment or orthognathic surgery, had no burns, injuries, or scar tissue in the head and neck regions, and had no breathing or swallowing disorder. The measurements of hard and soft tissues were carried out on lateral cephalometric radiographs taken at natural head position. The measurements were evaluated with a computer software program. The effects of sex on the measurements were investigated by means of Student's t test. RESULTS: The differences between men and women in nose prominence, basic upper lip thickness, upper lip thickness, inferior sulcus to the H line, and soft tissue chin thickness were statistically significant. CONCLUSION: The upper lip, nose, and chin are more protruded in men than they are in women. PMID- 12553620 TI - The effect of current-generation bonding systems on microleakage of resin composite restorations. AB - OBJECTIVE: The aim of this study was to determine the microleakage of current generation dentin bonding systems in Class II resin composite restorations. METHOD AND MATERIALS: Class II (occlusodistal or occlusomesial) cavity preparations with a gingival margin 2 mm apical to the cementoenamel junction were prepared on 70 noncarious, extracted human premolar teeth. Teeth were randomly divided into five groups and treated with different-generation bonding systems (Optibond FL, Gluma One Bond, Clearfil SE Bond, acid etching plus Clearfil SE Bond, and Prompt-L-Pop). All cavities were restored in a posterior resin composite and subjected to 200 thermocycles (at 5 degrees C to 55 degrees C; 30-second dwell time). After immersion in 0.5% basic fuchsin dye for 24 hours, the teeth were sectioned longitudinally and evaluated for dye penetration using a binocular stereomicroscope. RESULTS: There was no statistically significant differences among bonding systems in the degree of microleakage in the occlusal walls. For gingival walls, statistically significant differences were found only between the Clearfil SE Bond and Prompt-L-Pop groups and the Clearfil SE Bond with acid etching and Prompt-L-Pop groups. The greatest microleakage was observed in Prompt-L-Pop specimens. CONCLUSION: Most of the tested dentin bonding systems were able to eliminate microleakage completely in the occlusal walls, but some systems exhibited statistically significant differences in leakage in the gingival walls. PMID- 12553621 TI - Bond strength of a resin composite to a polyacid-modified resin composite under different conditions. AB - OBJECTIVE: The aim of this in vitro study was to evaluate the tensile bond strength values between polyacid-modified resin composite ("compomer") and resin composite materials under different conditions. METHOD AND MATERIALS: There were five experimental groups in the study. In group A, resin composite was placed directly on polyacid-modified resin composite surfaces. In group B, bonding agent was applied to polyacid-modified resin composite surfaces and then resin composite was placed on the compomer. In group C, compomer specimens were stored for 1 week and then resin composite was placed directly on these aged compomers. In group D, compomer specimens were again stored for 1 week, and then the bonding agent and resin composite were applied to the compomer surfaces. In group E, the surfaces of aged (1 week) compomers were roughened before the bonding agent was applied and the resin composite was placed on the prepared surfaces. A statistical analysis of the results was made with the Kruskal-Wallis test method. RESULTS: The mean tensile strength values of the groups were as follows: group A = 12.84 MPa; group B = 15.03 MPa; group C = 10.60 MPa; group D = 11.56 MPa; group E = 24.87 MPa. There were statistically significant differences between groups E and A; groups E and C; groups E and D; and groups C and B. CONCLUSION: Mechanical roughening of a polyacid-modified resin composite surface was found to be the most effective factor in increasing the tensile bond strength between an aged compomer and a resin composite. PMID- 12553622 TI - Unilateral masseter muscle hypertrophy: a case report. AB - Masseteric hypertrophy is a benign increase in the size of the masseter muscle, secondary to muscle hypertrophy. It produces facial asymmetry and is important in the differential diagnosis of other entities. The aim of this article is to report a case of unilateral masseteric muscle hypertrophy. The characteristics and diagnostic features of such alterations and options for treatment will be presented. PMID- 12553623 TI - Paradental cyst (inflammatory collateral, mandibular infected buccal cyst). PMID- 12553624 TI - Traction alopecia in children. AB - Traction alopecia is traumatic hair loss secondary to the application of tensile forces to scalp hair. This condition can be classified as marginal or nonmarginal. In either case, the induced trauma, often the result of cultural, social, and cosmetic practices, is unintentional. Initially, hair loss is reversible; however, permanent alopecia may result from chronic traction. PMID- 12553625 TI - What is your diagnosis? Dermatomyositis. PMID- 12553626 TI - What's eating you? Strongyloides stercoralis. PMID- 12553627 TI - Biologic therapy for psoriasis: the tumor necrosis factor inhibitors infliximab and etanercept. AB - During the past several years, one of the major focuses in psoriasis research has been the development of novel biologic therapies for this disease. The aim of these therapies is to provide selective, immunologically directed intervention, with the hope that such specificity will result in fewer side effects than traditional therapies. In this 2-part review, we present an update on the progress of the 4 biologic agents that most likely will be the first available for clinical use: infliximab, etanercept, efalizumab, and alefacept. The structure and mechanism of each drug will be reviewed, as well as the most recent clinical experience and safety data. The first article of this review will focus on the therapies that inhibit tumor necrosis factor alpha (TNF-alpha). PMID- 12553628 TI - Topical therapy with tretinoin and ammonium lactate for acanthosis nigricans associated with obesity. AB - Acanthosis nigricans (AN) is a cutaneous marker of various underlying systemic conditions. To date, no satisfactory topical therapy for this cutaneous disorder has been described. The following is a report of the successful use of a combination of 12% ammonium lactate cream and 0.05% tretinoin cream to treat AN associated with obesity. PMID- 12553629 TI - Biologic therapy for psoriasis: the T-cell-targeted therapies efalizumab and alefacept. AB - During the past several years, a new generation of therapies for psoriasis has been in development. These biologic therapies target the activity of T lymphocytes and cytokines responsible for the inflammatory nature of this disease. The first article of this 2-part update reviewed the tumor necrosis factor (TNF) inhibitors, infliximab and etanercept. In this article, we will review 2 therapies that target the T cell, efalizumab and alefacept. PMID- 12553630 TI - Chondroid syringoma. AB - We present the case of a 25-year-old man who presented with a slowly enlarging nodule on the upper lip. This nodule first appeared after minor trauma to the area. Our original differential diagnosis included foreign body granuloma, mucocele, inclusion cyst, and adnexal tumor. Histopathology results showed an encapsulated tumor consisting of 2 discrete lobules composed of numerous aggregates of cuboid epithelial cells with tubuloalveolar structures and keratinous cysts within a chondroid stroma-consistent with a chondroid syringoma. We compare and contrast the clinical and histologic characteristics of chondroid syringomas (arising from sweat glands) with those of pleomorphic adenomas (arising from salivary glands). PMID- 12553631 TI - Recalcitrant tinea corporis as the presenting manifestation of patch-stage mycosis fungoides. AB - Mycosis fungoides is a cutaneous T-cell lymphoma. Its presence, which denotes an altered immune system, may make treatment of otherwise simple cutaneous infections difficult. In the case presented here, a patient with widespread tinea corporis poorly responsive to several oral antifungals was noted as having a background poikilodermatous slightly scaly eruption. Results of a skin biopsy during therapy with oral antifungal medications showed evidence of tinea corporis; atrophy of the epidermis; a superficial, perivascular, and interstitial lymphocytic infiltrate with numerous atypical lymphocytes; and exocytosis of atypical lymphocytes into the epidermis with formation of microabscesses-findings consistent with the diagnosis of mycosis fungoides. Treatment with PUVA (oral psoralen and UVA light) and oral itraconazole led to long-term remission of the mycosis fungoides and the associated tinea corporis. Immune suppression may have contributed to the recalcitrant nature of our patient's dermatophyte infection. Underlying cutaneous, systemic, or iatrogenic disorders associated with immune dysfunction should be considered in patients with recalcitrant dermatophyte infections. PMID- 12553632 TI - Stress in a case of SAPHO syndrome. AB - In this article, we describe a case of seemingly stress-induced SAPHO (synovitis, acne, pustulosis, hyperostosis, osteitis) syndrome in a man who exhibited the central features of palmoplantar psoriasis and anterior chest involvement. We also review the etiology, pathogenesis, and treatment of SAPHO syndrome and emphasize the important differences between this syndrome and psoriatic arthritis. PMID- 12553633 TI - Treatment of compensatory hyperhidrosis with botulinum toxin type A. AB - Compensatory hyperhidrosis is a common and potentially severe side effect of bilateral endoscopic thoracic sympathectomy. We describe a patient with severe compensatory hyperhidrosis as a result of this procedure. We treated the patient with 2 separate series of subcutaneous botulinum toxin type A injections to the right upper abdomen and achieved anhidrosis in 5 weeks. Although botulinum toxin is already an established treatment modality for primary hyperhidrosis, there is little experience using it for the treatment of compensatory hyperhidrosis. Because of its efficacy and low side-effect profile, we believe botulinum toxin may play a significant role in treating compensatory hyperhidrosis as a first line agent. PMID- 12553634 TI - Sun protection education in the United States: what we know and what needs to be taught. AB - The incidence of skin cancers in the United States is rising. This has correlated with a life-time of sun exposure and cumulative damage of repetitive sun-related injuries such as tanning and sunburning. It is estimated that 80% of sun damage occurs before the age of 18 years. This relates to excessive tanning, blistering sunburns, and ineffective sun protection. It has been demonstrated that children can be taught to protect themselves from the sun. However, teaching of sun protection needs to begin at an early age. Such education requires that parents enforce protection in the household and through their actions in sunny situations. It has become apparent from the literature in the United States that we have made strides in sun education, but we have not yet instituted an effective nationwide education plan. This article reviews the data we have in the United States on sun protection, with a focus toward helping to design better education programs for the future. Based on the literature, it would seem that sun education should combine teaching mothers about sun protection in the nursery and teaching schools how to educate youngsters on the need for sun protection, beginning in nursery school. Improvements in education will have a latency of many years for reducing skin cancer incidence but will be an excellent investment in the future cutaneous health of today's children. PMID- 12553636 TI - Childhood herpes zoster. PMID- 12553635 TI - A double-blind clinical trial comparing the efficacy and safety of pure lanolin versus ammonium lactate 12% cream for the treatment of moderate to severe foot xerosis. AB - Xerotic skin is a pattern of reaction to a variety of disorders (eg, winter xerosis, hereditary ichthyosis) with abnormalities of desquamation in common. The trial described in this article was a double-blind randomized-comparison clinical trial investigating the effect of pure lanolin versus ammonium lactate 12% cream in treating moderate to severe foot xerosis. Xerosis was clinically assessed at baseline visit, and designated sites were reevaluated after 1, 2, and 4 weeks of therapy. Ninety-two patients were enrolled; 41 of these were excluded from analysis (25 were lost to follow-up, 8 were noncompliant, 1 was discharged from study because of an adverse event after visit 1, and 7 were noncompliant after visit 2). Although there was significant improvement in xerosis scores for both treatment groups after 2 and 4 weeks of treatment, no differences were statistically significant. Used twice daily for 4 weeks, pure lanolin and ammonium lactate cream were both effective in treating moderate to severe xerosis. PMID- 12553637 TI - Personal awareness of domestic violence: implications for health care providers. AB - OBJECTIVES: To estimate how many Californians know a victim of domestic violence, to examine their knowledge of certain characteristics of the violence, and to estimate and examine such knowledge among specific demographic groups. METHOD: A total of 3713 California adults (similar numbers of whites, blacks, Hispanics, Korean Americans, Vietnamese Americans, and other Asian Americans) completed a random-digit-dial interview. Respondents were asked whether a friend, relative, or coworker had been threatened or harmed by an intimate partner. Weights were applied to the cross-sectional sample to obtain estimates for the general population. Descriptive statistics and multivariate regressions were used with the full sample. RESULTS: Nearly half (45.5%) of the adult general population of California knows a victim of domestic violence (DV); 40.5% know a woman and 5.0% know a man. More than one-third of the population (35.7%) knew the victim while the abuse was happening. Although 86.5% of those who reported knowing a DV victim indicated that the victim incurred physical harm, only 18.3% of the injured victims were reported to have sought medical care. Gender was the most consistent respondent predictor: Men were less likely to know someone who was a victim of DV and to have specific information about the violence. Ethnic differences were fewer, but distinctions among groups were documented. CONCLUSIONS: Knowing a victim of DV is common among California adults. Implications for medical practice are discussed in terms of the ethnicity, gender, and work force status of the population served and geographic location of the medical practice. PMID- 12553638 TI - Health and professional characteristics of lesbian and heterosexual women physicians. AB - OBJECTIVE: To determine whether lesbian and heterosexual female physicians differ on health, professional, and demographic characteristics. METHODS: The Women Physicians' Health Study (WPHS), a mailed questionnaire sample survey, was completed by 4501 women physicians (59% response rate), of whom 115 were identified as lesbians and 4177 were identified as heterosexuals. Lesbian and heterosexual were defined by response to items about self-identification and sexual behavior. Dependent variables are self-reported health status and behavior and professional characteristics. RESULTS: Lesbians weighed more and were more likely to report histories of depression or sexual abuse, family histories of alcoholism, and orientation-related workplace harassment (all p = .01). On the other hand, they were more likely to comply with mammogram screening (p < .01). CONCLUSIONS: Many health status and behavior differences found in other studies of lesbian and heterosexual women were less pronounced here. This may be because the population of female physicians is more homogeneous, but perhaps also because the size of our lesbian sample was limited (n = 115). However, even with the buffer of high socioeconomic status and medical education, lesbian physicians seem to have a somewhat higher cumulative risk of disease compared to their heterosexual female colleagues. PMID- 12553639 TI - Female intimate partner homicide: a population-based study. AB - OBJECTIVES: To identify the patterns of female intimate partner homicide, to describe the nature of the partner relationships between victims and perpetrators, and to determine the extent of intimate partner violence (IPV) preceding the homicide. METHODS: We reviewed North Carolina medical examiner records and conducted telephone interviews with law enforcement officers about partner homicides of women age 15 and older occurring in North Carolina from 1991 to 1993. RESULTS: Most women were killed in homes (80%) and with guns (66%). One hundred thirty-five women (of 293 cases) were killed by marital partners, 47 of them former partners. Nonmarital partners killed 158 women, 46 of them former partners. Intimate partner violence was noted in two-thirds of the cases, 70% of those involving marital partners and 64% involving nonmarital partners. Homicides by former nonmarital partners were the most likely to have been preceded by IPV (78%). One hundred six victims with histories of IPV had had contact with law enforcement or judicial systems in the year before death. CONCLUSIONS: Legal and health care professionals should be aware that women are vulnerable to both IPV and homicide from both nonmarital and former partners as well as from current husbands. PMID- 12553640 TI - Planning your career in academic medicine. PMID- 12553641 TI - The hormone therapy dilemma: women respond. AB - OBJECTIVE: In the wake of the premature end of the Women's Health Initiative (WHI) study, we sought to assess women's knowledge of and attitudes about hormone therapy (HT) study findings and to appraise women's responses and intentions. METHODS: Between July 26 and August 6, 2002 a national random-digit-dialing telephone survey was conducted in a sample of households that included women 40 to 79 years old. RESULTS: Sixty-four percent of the 819 women interviewed had heard something about HT study results from the media or from talking with others, and 74% were confused about HT use. Another 57% were worried about how the findings might affect them, and 79% were interested in obtaining additional information about HT Only 24% of those who had heard something had actually sought additional information. Logistic regression findings suggested that women currently taking HT were most likely to be aware and informed. They also were more likely to be confused, worried, or to need or to seek additional information. Older women were less likely to be confused or worried or to need or seek additional information. More highly educated women were more likely to be aware and less likely to be confused or uninformed, but were more likely to have sought additional information. CONCLUSION: The Women's Health Initiative study provided a clear message about health risks and benefits of HT use. An important next step is to continue to convey accurate information to women, health providers, and the media so that women can make informed decisions about HT. PMID- 12553642 TI - Antecedents of euthanasia and suicide among older women. AB - OBJECTIVES: To identify the characteristics of older women who sought Jack Kevorkian's assistance in dying and to compare them with those of an age-matched sample who committed suicide. METHOD: This retrospective case-control study compared all 18 women age 55 and older who died with the assistance of Jack Kevorkian and whose deaths were investigated in Oakland County, Michigan from 1995 to 1997 with all 15 women age 55 and older who committed suicide in the same county during the same time period. We coded 203 variables in 7 domains from medical examiner files, including autopsy findings. RESULTS: Significantly more of Kevorkian's cases had amyotrophic lateral sclerosis or multiple sclerosis (p = .018), a recent decline in health (p = .031), or inadequately controlled pain (p = .041). Women who committed suicide had more prevalent chronic illnesses and were more likely to have been diagnosed with clinically significant depression or other psychiatric disorders (p = .023). Both groups were significantly less likely to be married (p < .001) and more likely to be divorced (p < .001) than US Census data would predict. CONCLUSIONS: The different vulnerabilities of older women who want to die and either commit suicide or seek assistance deserve continued careful research. Poorly controlled pain was a factor in seeking assistance in dying, and depression and psychiatric disorders characterized older women who committed suicide in our study. Not having a spouse may increase isolation and reinforce the hopelessness of women who are living with catastrophic illness. PMID- 12553643 TI - Considerations about TENORM: a study case on niobium facilities. AB - Two niobium mining facilities located at two different geological settings were selected as case studies for assessing the radiological impacts associated with mining and milling activities. The impacts were assessed both for operational and post-operational (future unrestricted use of the area) scenarios. The work methodology encompassed sampling and analysis of different materials along the operational process, including wastes, effluents and by-products, mass balance calculations, and dose assessment. The exposure scenario considered during the operational phase included the land use by hypothetical groups of members of the general public. In the case of post-operational phase the possible occupation of contaminated areas was considered. The results pointed out that during the operational phase of the investigated industries no relevant impacts could be attributed to them. However, in the case of the potential occupation of the contaminated areas where the wastes are being deposited, the unrestricted use of these areas cannot be accepted in terms of the relevant radiation protection requirements. PMID- 12553644 TI - Anomalous 235U/238U ratios and metal elements detected in the black rain from the Hiroshima A-bomb. AB - Contents of black rain from the A-bomb are important to clarify the health and environmental hazard caused by nuclear warfare. We analyzed the only existing physical remains of the black rain from Hiroshima A-bomb (Little Boy), a plastered wall with black streaks. Excess 137Cs and anomalous 235U/238U ratios detected in the black residues on the wall confirmed that the black streaks on the wall originated from Little Boy. The results also showed that excess 235U could be used as a fingerprint to clarify the distribution of materials discharged from Little Boy into the environment. High concentrations of lead and zinc, which may have caused a degree of heavy metal pollution in the area where the rain fell, were also detected in the black rain sample. PMID- 12553645 TI - Exposure to oral uranyl nitrate delays tooth eruption and development. AB - The risk of oral exposure to uranium potentially involves the population at large. Tooth eruption and development are ongoing processes that begin during fetal development and continue until the age of 18 y. Since one of the mechanisms involved in tooth eruption is bone formation and it is well documented that uranium inhibits bone formation, the aim of the present work was to study the effect of oral administration of uranyl nitrate (UN) on tooth eruption and development. Wistar rats aged 1 and 7 d were orally administered a single dose of 90 mg kg(-1) body weight of uranyl nitrate. Two age matched groups received an equal volume of saline and served as controls. The animals were killed at 7 and 14 d of age, respectively. Mandibles were resected and processed to obtain bucco lingual sections oriented at the level of the mesial root of the first mandibular molar, and histomorphometric studies were performed. Results showed that an acute high dose of uranyl nitrate delays both tooth eruption and development, probably due to its effect on target cells. PMID- 12553646 TI - Uptake of 137Cs and 90Sr in rice plants. AB - In order to study the transfer of water-borne 137Cs and 90Sr into rice plants, pot experiments were done by growing rice plants on soils taken from rice beds near two nuclear power plants: Qinshan Nuclear Power Plant and Daya Bay Nuclear Power Plant in China. The vertical migration of 137Cs and 90Sr in the soil samples was studied, and results show that almost all the contaminants were retained in the top 1 cm of soil with at most a few percent down to 3 cm of soil. The areal transfer factors of 137Cs and 90Sr were studied by adding various activity concentrations of the two radionuclides to the irrigation water and then measuring the activity concentration in the various rice tissues. The areal transfer factors were also analyzed for their dependence on soil characteristics, time and duration of the contamination. The areal transfer factors obtained in this study refer to the absorption of the fallout nuclides deposited in the irrigation water in a rice field as opposed to other studies that deal with uniform activity concentration in the soils. PMID- 12553647 TI - Measurements of accelerator-produced leakage neutron and photon transmission through concrete. AB - Optimum shielding of the radiation from particle accelerators requires knowledge of the attenuation characteristics of the shielding material. The most common material for shielding this radiation is concrete, which can be made using various materials of different densities as aggregates. These different concrete mixes can have very different attenuation characteristics. Information about the attenuation of leakage photons and neutrons in ordinary and heavy concrete is, however, very limited. To increase our knowledge and understanding of the radiation attenuation in concrete of various compositions, we have performed measurements of the transmission of leakage radiation, photons and neutrons, from a Varian Clinac 2100C medical linear accelerator operating at maximum electron energies of 6 and 18 MeV. We have also calculated, using Monte Carlo techniques, the leakage neutron spectra and its transmission through concrete. The results of these measurements and calculations extend the information currently available for designing shielding for medical electron accelerators. Photon transmission characteristics depend more on the manufacturer of the concrete than on the atomic composition. A possible cause for this effect is a non-uniform distribution of the high-density aggregate, typically iron, in the concrete matrix. Errors in estimated transmission of photons can exceed a factor of three, depending on barrier thickness, if attenuation in high-density concrete is simply scaled from that of normal density concrete. We found that neutron transmission through the high-density concretes can be estimated most reasonably and conservatively by using the linear tenth-value layer of normal concrete if specific values of the tenth-value layer of the high-density concrete are not known. The reason for this is that the neutron transmission depends primarily on the hydrogen content of the concrete, which does not significantly depend on concrete density. Errors of factors of two to more than ten, depending on barrier thickness, in the estimated transmission of neutrons through high-density concrete can be made if the attenuation is scaled by density from normal concrete. PMID- 12553648 TI - In situ calibration of a low-level, in-line, flow liquid scintillation analyzer. AB - A new method to calibrate a flow liquid scintillation detector in an in-line waste water monitor was developed and its operation was verified. The liquid scintillation detector for detecting low energy beta emitters was one of three detectors of the monitor that aimed to analyze radionuclide concentrations in the waste water before draining it into sewer. To calibrate the liquid scintillation detector, a 20-cm3 calibration vial containing a reference solution was connected to the water sampling channel with a three-way valve. Ten measurements needed to define a quench correction curve required only 200 cm3 solution. The counting efficiencies of the liquid scintillation detector for 3H and for 14C under the average quenching level of the waste waters were estimated at 27% and 58%, respectively. The calibration using the vial took into account the sampling and mixing system, which would not be the case with sealed quenched standards. Although active solution was used, the method was simple, generated little active waste, had low possibility of contamination, and reduced costs and labor for the calibration. PMID- 12553649 TI - An experimental determination of FIDLER scanning efficiency at specific speeds. AB - As part of mass remediation efforts across the country some radiation detection systems are now being used in conjunction with data logging and positioning system technology. These systems can be used in the scanning mode, simultaneously recording both count rate and position. Following data analysis, hot spots can be identified and remediation efforts for that particular area can commence. This technique has been used for nearly a decade and has had success in accelerating preliminary remediation work while also reducing potential clean up costs. However, little work has been completed on how the sensitivity of these detection systems are affected when used with this technology because while the intrinsic efficiency of the detector is constant, scanning efficiency can vary depending on data sampling time and scanning speed. To better understand scanning efficiency for a detector attached to such a system, a device was developed which moved soil at a constant speed underneath a Field Instrument for Detecting Low Energy Radiation (FIDLER). Count rate was measured every 2 s as a 241Am source passed under the detector at speeds ranging from approximately 10 cm s(-1) to 100 cm s( 1). A surface source and a buried source were both examined. Experimental detection efficiency was calculated and compared to Monte Carlo generated results. For the surface source, the efficiency dropped to a value of approximately 1% at 100 cm s(-1). At the same speed, the buried source had a detection efficiency of 0.1%, primarily due to attenuation of the low energy photon in the soil. It was also noted that the response time of the meter affected the scanning efficiency. With a response time set at 1 s, higher average efficiencies were recorded but with a large standard deviation from the mean. Higher response time setting had the effect of reducing the variability of the reading but also reducing efficiency. PMID- 12553650 TI - A comparison between gross and net count methods when searching for orphan radioactive sources. AB - Different numerical strategies in searching for orphan radioactive sources in the environment by means of a mobile detector system have been evaluated. A carborne 3- by 3-inch NaI(Tl) spectrometric system was used with an unshielded 2 GBq 137Cs source as a test source. In this paper, a previous method (MB method), based on a moving average algorithm applied on the gross count rate, was extended and compared with three moving average algorithm methods involving different natural background subtraction strategies. For each method the distance from the road that an orphan source can be detected with a probability of 50% (the critical distance, CD) when driving by is determined. The CD for a 2 GBq 137Cs source improved from 105 m to 130 m when the interference from 40K was subtracted from the 137Cs spectral window. However, when the contribution of other natural gamma emitting nuclides also was subtracted, the ability to find the 137Cs source was reduced. PMID- 12553651 TI - Measurements with a Ge detector and Monte Carlo computations of dose rate yields due to cosmic muons. AB - The present work shows how portable Ge detectors can be useful for measurements of the dose rate due to ionizing cosmic radiation. The methodology proposed converts the cosmic radiation induced background in a Ge crystal (energy range above 3 MeV) to the absorbed dose rate due to muons, which are responsible for 75% of the cosmic radiation dose rate at sea level. The key point is to observe in the high energy range (above 20 MeV) the broad muon peak resulting from the most probable energy loss of muons in the Ge detector. An energy shift of the muon peak was observed, as expected, for increasing dimensions of three Ge crystals (10%, 20%, and 70% efficiency). Taking into account the dimensions of the three detectors the location of the three muon peaks was reproduced by Monte Carlo computations using the GEANT code. The absorbed dose rate due to muons has been measured in 50 indoor and outdoor locations at Thessaloniki, the second largest town of Greece, with a portable Ge detector and converted to the absorbed dose rate due to muons in an ICRU sphere representing the human body by using a factor derived from Monte Carlo computations. The outdoor and indoor mean muon dose rate was 25 nGy h(-1) and 17.8 nGy h(-1), respectively. The shielding factor for the 40 indoor measurements ranges from 0.5 to 0.9 with a most probable value between 0.7-0.8. PMID- 12553652 TI - Comparison of sliced lungs with whole lung sets for a torso phantom measured with Ge detectors using Monte Carlo simulations (MCNP). AB - Lung counters are generally used to measure low energy photons (<100 keV). They are usually calibrated with lung sets that are manufactured from a lung tissue substitute material that contains homogeneously distributed activity; however, it is difficult to verify either the activity in the phantom or the homogeneity of the activity distribution without destructive testing. Lung sets can have activities that are as much as 25% different from the expected value. An alternative method to using whole lungs to calibrate a lung counter is to use a sliced lung with planar inserts. Experimental work has already indicated that this alternative method of calibration can be a satisfactory substitute. This work has extended the experimental study by the use of Monte Carlo simulation to validate that sliced and whole lungs are equivalent. It also has determined the optimum slice thicknesses that separate the planar sources in the sliced lung. Slice thicknesses have been investigated in the range of 0.5 cm to 9.0 cm and at photon energies from 17 keV to 1,000 keV. Results have shown that there is little difference between sliced and whole lungs at low energies providing that the slice thickness is 2.0 cm or less. As the photon energy rises the slice thickness can increase substantially with no degradation on equivalence. PMID- 12553653 TI - Should radon be reduced in homes? A cost-effect analysis. AB - BACKGROUND: Radon is a radioactive gas that may leak into buildings from the ground. Radon exposure is a risk factor for lung cancer. An intervention against radon exposure in homes may consist of locating homes with high radon exposure (above 200 Bq m(-3)) and improving these, and protecting future houses. The purpose of this paper is to calculate the costs and the effects of this intervention. METHODS: We performed a cost-effect analysis from the perspective of the society, followed by an uncertainty and sensitivity analysis. The distribution of radon levels in Norwegian homes is lognormal with mean = 74.5 Bq m(-3), and 7.6% above 200 Bq m(-3). RESULTS: The preventable attributable fraction of radon on lung cancer was 3.8% (95% uncertainty interval: 0.6%, 8.3%). In cumulative present values the intervention would cost $238 (145, 310) million and save 892 (133, 1981) lives; each life saved costs $0.27 (0.09, 0.9) million. The cost-effect ratio was sensitive to the radon risk, the radon exposure distribution, and the latency period of lung cancer. Together these three parameters explained 90% of the variation in the cost-effect ratio. CONCLUSIONS: The uncertainty in the estimated cost per life is large, mainly due to uncertainty in the risk of lung cancer from radon. Based on estimates from road construction, the Norwegian society has been willing to pay $1 million to save a life. This is above the upper uncertainty limit of the cost per life. The intervention against radon in homes, therefore, seems justifiable. PMID- 12553654 TI - Study of long term radon transport by measuring the difference of the 210Pb and 226Ra activity in soil as a function of the depth. AB - Long term radon transport has been studied by measuring the activity difference of 210Pb and 226Ra in soil as a function of the depth. The results are from test pits of 1-2 m depth made at a dam of a tailings pond and at the plateau of a waste rock pile. Soil samples of about 1 kg were taken at a successive distances of 5 cm and analyzed by means of gamma-ray spectroscopy using low background germanium n-type detectors. It is shown that by this method nearly model independent information about the radon transport over a period of several decades can be inferred from the data. PMID- 12553655 TI - Estimation of radiographic doses in a case-control study of acute myelogenous leukemia. AB - Radiation exposure from dental and medical radiography is inherently difficult to estimate. Because no single resource is available for this type of data, gathering the information needed to estimate exposure in epidemiologic studies is a labor-intensive, imprecise process. For a case-control study of adult-onset acute myelogenous leukemia in Los Angeles County and radiography, a database was created of estimates of dose to the red bone marrow for each radiographic procedure reported by subjects in interviews and recorded in subjects' medical records. Resources used included the medical literature as well as personal communications with radiology experts. Dose estimates for each procedure based on this database are reported. Methods and complications are contrasted with a past effort to estimate dose from dental radiography for a case-control study of parotid gland tumors. Among the more difficult aspects of medical radiography dose estimation are the wide variety of examinations performed, the continually changing environment of diagnostic imaging, and the number of variables that contribute to the delivered dose to an individual from a specific imaging procedure. PMID- 12553656 TI - Statistical validation of a commonly used method for personnel dosimetry issuance determinations. AB - In 1992, the federal requirements for personnel monitoring of occupational radiation doses were modified, making monitoring compulsory only for individuals "likely" to exceed 10% of the applicable annual dose limits. This regulatory change served as a catalyst for many radiation protection programs to re-evaluate their personnel monitoring issuance criteria to determine if some monitoring services might be reduced or eliminated. But in the absence of any clear definition of the term "likely," radiation protection programs relied on adjustments largely based on reviews of previous monitoring results and professional judgment. Although such semi-quantitative assessments may have been appropriate, the approach was not without inherent programmatic risks, as radiation dosimetry programs have been shown to be a consistent source of regulatory non-compliance upon inspection and serve as essential elements in defense against litigation efforts purporting workplace-related radiation injury. The objective of this study was to subject this commonly used method for post regulatory change dosimetry issuance determinations to statistical validation. Personnel monitoring data from The University of Texas Health Science Center at Houston Radiation Safety Program pre- and post-1992 regulatory changes were accessed and descriptively analyzed. The dynamic nature of the institutional environment made direct comparisons of individuals between years impractical, so 1990 mean dose levels for various independent variables were compared using the Kruskal-Wallis non-parametric ANOVA test at a significance level of alpha = 0.05. The data from 1998 were then analyzed to ascertain if the groupings identified as statistically different in 1990 were still being monitored. The analyses statistically validated the monitoring program reductions made by the University of Texas Health Science Center at Houston radiation protection program. In addition, statistical support for further monitoring program reductions were identified, if such reductions were deemed to be necessary in the future. PMID- 12553657 TI - Post-exposure medical surveillance of workers exposed to ionizing radiation. PMID- 12553658 TI - Optimisation of colon cleansing prior to computed tomographic colonography. AB - Colon preparation technique is a major determinant factor for patient compliance and polyp detection in computed tomographic colonography (CTC). The purpose of this study is to compare three different colon cleansing techniques in terms of patient discomfort, sensitivity and specificity. The following colon cleansing methods were compared in 20 patients each: 1. standard colonoscopy cleansing (ScCl) the day of the examination, based on polyethylene glycol (PEG), 2. a slightly reduced cleansing (RcCI) the day prior to the examination, based on a combination of diet, bisacodyl and a reduced intake of PEG, and 3. a cleansing with dietary fecal tagging (FT) the day prior to the examination, based on a combination of diet, bisacodyl, magnesium citrate and a dedicated barium suspension. ScCl resulted in a clean colon, but produces fluid levels hampering a complete CTC and possibly resulting in false negative diagnosis. RcCl reduced the problem of fluid levels, but was faced with the problem of fecal residues, resulting in false positive diagnosis. FT offered the possibility to obtain a dry colon, with labelled fecal residues, thus reducing false positive findings. Optimisation of the diet and replacement of PEG by magnesium citrate in FT reduced the preparation related discomfort and improved the final opinion. FT is the preferred colon cleansing technique because, compared to ScCl, fluid levels are reduced, and compared to RcCl, differentiation between faecal residues and polyps is improved. Moreover, FT reduces preparation related discomfort, compared to both RcCl and ScCl. PMID- 12553659 TI - Solid and papillary epithelial neoplasm of the pancreas: MR imaging findings. AB - We present the magnetic resonance imaging (MRI) findings in a young patient with histological proven solid and papillary epithelial neoplasm of the pancreas. Although the appearance of this tumor at MRI can be useful in making a correct preoperative diagnosis, only few cases describing its MRI features have been reported in the radiological literature. PMID- 12553660 TI - Bilateral supernumerary kidney: imaging findings. AB - Supernumerary kidney is a rare urogenital anomaly with less than 100 cases reported. Bilateral supernumerary kidney is even more rare and to our knowledge there are three cases reported in the literature. Herein we present the computed tomography (CT), excretory urography (IVP), and CT angiography (CTA) findings in a patient with bilateral supernumerary kidneys. PMID- 12553661 TI - Chronic pancreatitis: diagnostic role of computed tomography and magnetic resonance imaging. AB - The value of computed tomography (CT) and magnetic resonance imaging (MRI) in the diagnosis and detection of complications in patients with chronic pancreatitis are reviewed. CT and MRI diagnoses are based on changes in the pancreatic duct and parenchyma and on the detection of ductal calculi. Despite technical refinements of these imaging methods, the diagnosis of chronic pancreatitis remains often difficult because of the complexity of the morphologic changes and the false-negative results in the early stages of the disease. This article describes and illustrates the imaging features of chronic pancreatitis classified according to the underlying etiology. PMID- 12553662 TI - Spinal intramedullary arteriovenous malformation. PMID- 12553663 TI - The body packer syndrome. PMID- 12553664 TI - A novel phospholipid-binding protein from the yeast Saccharomyces cerevisiae with dual binding specificities for the transport GTPase Ypt7p and the Sec1-related Vps33p. AB - The gene product of the Saccharomyces cerevisiae open reading frame YDR229w (named IVY1 for: Interacting with Vps33p and Ypt7p) was found to interact with both the GTPase Ypt7p and the Sec1-related Vps33 protein. While deletion of IVY1 does not lead to any recognized change in phenotype, overexpression of Ivy1p leads to fragmentation of the vacuole, missorting of the vacuolar enzyme carboxypeptidase Y (CPY) to the exterior of the cell, and an accumulation of multivesicular bodies inside the cell. All effects caused by the overexpression of Ivy1p can be reset by simultaneously raising the amount of Vps33p. This suppression activity of Vps33p suggests that Ivy1p and Vps33p at least partially counteract the action of each other in the cell. The intracellular level of Ivy1p increases in cells approaching stationary growth phase at which part of the protein is located at the rim of the vacuole. In addition to its specific interactions with members of two regulatory protein families, Ivy1p in vitro shows a marked propensity for binding phospholipids with high affinity. PMID- 12553665 TI - Tyrosine-based endocytic motifs stimulate oligomerization of AP-2 adaptor complexes. AB - The clathrin adaptor complex AP-2 functions in the assembly of clathrin-coated vesicles at the plasma membrane where it serves to couple endocytic vesicle formation to the selection of membrane cargo proteins. Recent evidence suggests that binding of tyrosine-based endocytic sorting motifs may induce a conformational change within the AP-2 adaptor complex that could enhance its interaction with other cargo molecules and with the membrane. We report here that soluble tyrosine-based endocytic sorting motif peptides facilitate clathrin/AP-2 recruitment to liposomal membranes and induce adaptor oligomerization even in the absence of a lipid bilayer. These effects are specific for endocytic motifs of the type Yxxphi whereas peptides corresponding to NPxY- or di-leucine-containing sorting signals are ineffective. Our data may help to explain how the highly cooperative assembly of clathrin and adaptors could be linked to the selection of membrane cargo proteins. PMID- 12553666 TI - Importance of the propeptide in the biosynthetic maturation of rat cathepsin C. AB - Cathepsin C is a cysteine dipeptidyl-aminopeptidase. Active cathepsin C is found in lysosomes as a 200-kDa multimeric enzyme. Subunits constituting this assembly all arise from the proteolytic cleavage of a single precursor giving rise to three peptides: the propeptide, the alpha- and the beta-chains. Some features of the propeptide such as its length, its high level of glycosylation and its retention in the active lysosomal form of the enzyme suggest an important contribution of the proregion in the transport, maturation and expression of cathepsin C. In order to assess some aspects of this contribution, we transiently expressed mutant molecules of rat cathepsin C either lacking three of the four glycosylation sites, partially deleted in the proregion, or mutated at tryptophan 39 also located in the proregion, and studied their biosynthesis. Our results show that at least one of the three glycosylation sites in the propeptide must be glycosylated in order to obtain targeting and maturation of cathepsin C. We also show that a deletion of 14 amino acids and mutation W39S in the propeptide totally abolishes the biosynthetic processing of the enzyme. These results demonstrate that in addition to its role as a chaperone or in maintaining the latency of the enzymatic activity, the propeptide is required for proper transport and expression of newly synthesized cathepsin C. PMID- 12553667 TI - The secretory beta-amyloid precursor protein is a motogen for human epidermal keratinocytes. AB - Cell migration is known to be triggered by constituents of the extracellular matrix such as fibronectin and by soluble mediators commonly summarized as motogens. Many growth factors such as the epidermal growth factor (EGF) have been shown to act as motogens. Recently, the secretory N-terminal portion of the beta amyloid precursor protein (sAPP) has been identified as a keratinocyte growth factor. Hence, in this study we analysed whether sAPP stimulates also keratinocyte migration employing the stroboscopic cell motility assay. The migration velocity as well as the frequency of lamellipodia protrusion and ruffle formation were increased about two-fold thus corresponding to the effect of EGF. Using a newly developed beta1-integrin migration track assay we observed that sAPP increased the proportion of migrating keratinocytes and their directional persistence. sAPP appeared to operate synergistically with fibronectin with respect to its motogenic effect. Using a modified Boyden chamber assay we showed that sAPP besides its chemokinetic effect functions as a chemoattractant. Like EGF, sAPP exerted its motogenic effect through the activation of Rac kinase but the receptor for sAPP appears to be distinct. The results suggest that sAPP operates as a motogen in the human epidermis, where it may participate in the regulation of reepithelialization during wound healing. PMID- 12553668 TI - Partial cleavage of A-type lamins concurs with their total disintegration from the nuclear lamina during apoptosis. AB - Although activated caspase 6 is capable of cleaving both A- and B-type lamins during apoptosis, the higher-order structure of the nuclear lamina may cause a differential breakdown of these two types of lamins. In order to obtain a better understanding of the dynamics and the consequences of the rapid, coordinated breakdown of the lamina complex, we applied the green fluorescent protein (GFP) technology in living cells, in which the fate of individual caspase cleavage fragments of A- and B-type lamins was examined. CHO-K1 cells were stably transfected with cDNA constructs encoding N-terminally GFP-labelled hybrids of lamin A, lamin Adelta10, lamin C or lamin B1. The course of the apoptotic process, induced by the kinase inhibitor staurosporine or by the proteasome inhibitor MG132, was monitored by digital imaging microscopy or confocal microscopy. Time-lapse recordings showed that parallel to DNA condensation N terminally GFP-tagged A-type lamins became diffusely dispersed throughout the nucleoplasm and rapidly translocated to the cytoplasm. In contrast, the majority of GFP-lamin B1 signal remained localised at the nuclear periphery, even after extensive DNA condensation. Comparison of lamin B1-GFP signal with A-type lamin antibody staining in the same apoptotic cells confirmed the temporal differences between A- and B-type lamina dispersal. Immunoblotting revealed only a partial cleavage of A-type lamins and an almost complete cleavage of lamin B1 during apoptosis. In contrast to lamin B1 in normal cells, this cleaved lamin B1, which is apparently still associated with the nuclear membrane, can be completely extracted by methanol or ethanol. Fluorescence loss of intensity after photobleaching experiments showed that in apoptotic cells A-type lamin-GFP molecules diffuse almost freely in both nucleoplasm and cytoplasm, while the lamin B1-GFP fragments remain more stably associated with the nuclear membrane, which is confirmed by co-localisation immunofluorescence studies with a nucleoporin p62 antibody. Our results therefore clearly show a differential behaviour of A- and B-type lamins during apoptosis, suggesting not only distinct differences in the organisation of the lamina filaments, but also that caspase cleavage of only a small fraction of A-type lamins is needed for its complete disintegration. PMID- 12553669 TI - Vimentin intermediate filament reorganization by Cdc42: involvement of PAK and p70 S6 kinase. AB - Rho family GTPases play a major role in actin cytoskeleton reorganization. Recent studies have shown that the activation of Rho family GTPases also induces collapse of the vimentin intermediate filament (IF) network in fibroblasts. Here, we report that Cdc42V12 induces the reorganization of vimentin IFs in Hela cells, and such reorganization is independent of actin and microtubule status. We analyzed the involvement of three serine/threonine kinase effectors, MRCK, PAK and p70 S6K in the Cdc42-induced vimentin reorganization. Surprisingly, the ROK related MRCK is not involved in this IF reorganization. We detected phosphorylation of vimentin Ser72, a site phosphorylated by PAK, after Cdc42 activation. PAK inhibition partially blocked Cdc42-induced vimentin IF collapse suggesting the involvement of other effectors. We report that p70 S6 kinase (S6K)1 participates in this IF rearrangement since the inhibitor rapamycin or a dominant inhibitory S6K could reduce the Cdc42V12 or bradykinin-induced vimentin collapse. Further, inhibition of PAK and S6K in combination very effectively prevents Cdc42-induced vimentin IF collapse. Conversely, only in combination active PAK and S6K could induce a vimentin IF rearrangement that mimics the Cdc42 effect. Thus, Cdc42-induced vimentin reorganization involves PAK and, in a novel cytoskeletal role, p70 S6K. PMID- 12553670 TI - Sentinel lymph node--its imaging and detection in breast carcinoma. PMID- 12553671 TI - Usefulness of K-1 (CD-30) marker in Hodgkin's disease. AB - OBJECTIVE: To identify Hodgkin's and Reed Sternberg like cells by a single Immunomarker (Ki-1; CD-30) and to highlight diagnostic specificity of these cells in Hodgkin's disease, as these cells are frequently encountered in other lymphoproliferative disorders on H&E sections. METHODOLOGY: Seventy-nine histologically diagnosed cases of lymph node biopsies (59 cases of Hodgkin's disease, 15 cases of Non Hodgkin's lymphoma and 5 cases of non neoplastic lymphoid tissue) were subjected to Ki-1 (CD-30) immunostaining as all cases revealed Hodgkin's and/or Reed Sternberg like cells on H&E stained sections. RESULTS: Out of 59 cases of Hodgkin's disease, 49 (83%) showed Ki-1 (CD-30) immunoreactivity for Reed Sternberg and Hodgkin's cells. None of the 15 cases of non-Hodgkin's lymphoma (2 small lymphocyte type, 11 diffuse large cells type, 2 lymphoblastic type) showed positive Ki-1 (CD-30) immunostaining. Only one case of the remaining 5 cases of non-neoplastic lymphoid lesion (2 toxoplasmic lymphadenitis, one histiocytic necrotizing lymphadenitis, one reactive hyperplasia and one chronic non specific lymphadenitis) showed positive Ki-1 immunostaining. CONCLUSION: Ki-1 (CD-30) is a reliable marker for Hodgkin's Reed Sternberg like cells in histologically proven cases of Hodgkin's disease but not for similar cells encountered in other lymhoproliferative lesions. PMID- 12553672 TI - Impact of weather on presentation of kidney tumour: a thirteen years retrospective study. AB - OBJECTIVE: Presentation of kidney tumor patients in-group gave birth to the hypothesis that oncogen exposure is influenced by the weather. Frequency and spectrum of renal tumors was also studied. Incidence of the tumor cannot be evaluated as we are lacking in data collection of pediatric population, inoperable cases and patients with bilateral renal tumors, who did not under go nephrectomies. SUBJECTS AND METHODS: Case records of all patients who underwent nephrectomy at the Department of Urology, Pakistan Institute of Medical Sciences (PIMS), Islamabad, between 1988 and 2000 were retrospectively analyzed. Histologically proven cases of renal cancer seen over 13 years period under review, were included in the study. RESULTS: Between 1988 to 2000, 80 histologically proven cases of renal carcinoma were seen. The ages of the patients ranged from 10 to 80 years (median 55 years) with a male to female ratio of 2:1. The frequency of kidney tumor in different months was evaluated. Spring and end of summer season showed increase number of kidney tumor patients 27.50% and 30% respectively but it is not significant. This is a small study at one center and data from other centers may be conclusive. Histology showed renal cell carcinoma in 86.2%, squamous cell carcinoma in 3.75%, transitional cell carcinoma in 3.75%, Wilms' tumor and Non-Hodgkin's lymphoma in 1.25% each and undifferentiated in 2.5% cases. Invasiveness was found in 16.25% cases, 7.5% had renal vein involvement, 6.25% lymph node and 2.5% inferior vena cava. Side of involvement was right in 51.2% and left in 48.8% cases. Of the total urological malignancies that were seen during 13 years period under review, renal carcinoma was found in 12.4% cases. Of the total nephrectomies that were carried out during this period 24.5% were the cases of renal carcinoma. Hospital frequency of renal carcinoma was 2.28/100,000/year or 6.1 tumors per year. CONCLUSION: Impact of weather could not be proved a causative factor for kidney tumor presentation. This study also indicates that although the frequency of renal carcinoma may be different from western countries but the spectrum of the disease is quite similar. PMID- 12553673 TI - An evaluation of cefaclor in Pakistani children with pharyngotonsillitis. AB - OBJECTIVE: To assess the efficacy, safety of cefaclor for the treatment of pharyngotonsillitis in Pakistani children. METHOD: Pakistani children between the ages of two to twelve years and diagnosed with pharyngotonsillitis were eligible to participate in the study. Cefaclor was administered three times daily for a total dose of 20-40 mg/kg/day, depending on the severity of the infection, for 7 days. Patients were evaluated on the basis of symptomatic response and antibiotic susceptibility pattern as a result of bacteriological examination. RESULTS: Based on observed pre-therapy signs/symptoms of infection with pharyngotonsillitis, 99% of patients (95% CI=94.9% to 100.0%) were reported as having a symptomatic response of Cure or Improvement by end of study. Group A b-haemolytic Streptococci was the most prevalent pathogen isolated pre-therapy (occurring in 45% of patients). Of all patients for whom this pathogen was isolated, 64% had complete eradication by end of therapy, while there was persistence or superinfection in 36% of patients. Only 9 occurrences of non-serious adverse events were observed among 7 of the 107 patients, including diarrhea (5), nausea (1), vomiting (1), rash (1) and stomatitis (1). CONCLUSION: Based on the high symptomatic response rate of 99% and the positive safety data, cefaclor was found to be effective and safe in treating Pakistani children with pharyngotonsillitis. PMID- 12553674 TI - Clinical and demographic characteristics of psychiatric inpatients admitted via emergency and non-emergency routes at a university hospital in Pakistan. AB - OBJECTIVE: To examine the differences in the demographic and clinical characteristics of patients admitted through emergency versus non-emergency routes and see if these two groups of patients were significantly different from each other with respect to criteria mentioned in the title. METHODS: Retrospective data was analysed in all 2576 patient records were reviewed and these patients were divided in two groups with respect to their mode of admission (emergency vs non emergency). These groups were then compared with respect to sex, age, length of stay, discharge status and diagnostic categories. Statistical package for social sciences version 8.0 (SPSS 8.0) was used to analyze the data. SETTING: The study was conducted at the Aga Khan University Hospital, is a private tertiary care hospital with a 13 bed psychiatric facility. RESULTS: When these two groups were compared, significant differences were found, with ER patients having a shorter length of stay and youngest mean age, proportion of females admitted via ER was greater than those in non-ER group. There were significantly more women in each group who were married. The percentage of patients who left against medical advice was greater in the ER group. In both the groups mood disorders including Bipolar disorder and major depressive disorders was the most prevalent category with psychotic disorders to follow. The percentage of patients in both these categories was greater in the non-ER group where as percentage of patients with conversion disorder was higher in the ER group. CONCLUSION: Significant differences were found in the patient characteristics admitted via ER versus Non ER. Shorter length of stay in ER group might indicate an acute episode resolving quickly. Studies need to be done prospectively to determine the difference in the two groups thus ascertaining the level of care needed for each group. Lastly, the high proportion of patients coming through the ER also indicates that there exists a need for primary care involvement in mental health care thus reducing the need for emergency room usage. Clinical and Demographic Characteristics of inpatients admitted via emergency and non-emergency routes at a university hospital in Pakistan. PMID- 12553675 TI - Comparison of fentanyl and nalbuphine in total intravenous anaesthesia (TIVA). AB - OBJECTIVE: To compare Nalbuphine and fentanyl as total intravenous anaesthesia with propofol infusion in laproscopic cholecystectomy cases. STUDY DESIGN: Double blind randomised. METHODS: Changes in haemodynamic variables greater than twenty percent above or below the baseline and recovery profile were observed. RESULTS: Blood pressure remained within 20% of baseline in either group. Nine patients in fentanyl and fifteen in nalbuphine group required an additional bolus of propofol intraoperatively. Heart rate response after tracheal intubation was significantly higher in the nalbuphine group (25%). No difference was observed in the incidence of nausea and vomiting in the recovery room. Twenty-seven percent patients in the nalbuphine group required analgesia in the recovery in comparison to 87% in the fentanyl group. Patients in the fentanyl group required analgesia earlier (37 minutes vs. 62 minutes). CONCLUSION: Fentanyl provided better intraoperative haemodynamic stability in comparison to nalbuphine when used as the analgesic component in total intravenous anaesthesia with propofol. The recovery profile with both drugs was similar. Lesser number of patients required analgesia in the recovery in the nalbuphine group. PMID- 12553676 TI - Diabetes related knowledge, attitude and practices of family physicians in Pakistan. AB - OBJECTIVE: To assess the Knowledge, Attitude and Practices (KAP) towards diabetes of Family Physicians (FPs) working in urban and rural areas of Pakistan. METHODOLOGY: A cross-sectional survey was conducted on FPs through an interview by a trained person and filling up a questionnaire focused on diagnosis, treatment and complications of diabetes. The answers were scored by assigning marks and conversion to percentages. The computer package SPSS version 10 was used for statistical analysis. RESULTS: A total of 767 FPs (756 males, 11 females, average age 42.18 years) with a mean clinical practice duration of 13.41 years, with 681 practicing in urban areas and 86 in rural areas, participated in the study. Each FP saw on an average 58 patients daily, spending a mean of 8.5 minutes with each case. More than 90% FPs treated diabetics and 107 (14%) of the doctors were diabetic themselves. Overall 62% answers were correct, with the FPs from Sindh having the highest score of 66% and the Federal Capital Area with the lowest 54%. The questions answered correctly by less than 50% of FPs pertained to diagnostic blood values of glucose, treatment of children with diabetes, pregnant diabetics, monitoring of diabetics and technique of insulin injection. The questions answered correctly by more than 50% of FPs were related to diagnosis of Impaired Glucose Tolerance (IGT), insulin in pregnancy, importance of education in diabetics and diabetes complications, screening and management. CONCLUSION: This study has explored several aspects of diabetes related KAP of Family Physicians and identified the need for improvement in their practices for treating and educating diabetics. Awareness and education programmes are recommended to update the FPs on early detection and management of diabetes. PMID- 12553677 TI - Outcome of singleton breech deliveries beyond 28 weeks gestation: the experience at MCH Centre, PIMS. Mother and Child Health. Pakistan Institute of Medical Sciences. AB - OBJECTIVE: To conduct an audit of breech deliveries at or beyond 28 weeks gestation to determine the neonatal outcome at different gestations in relation to the mode of delivery. SETTING: MCH Centre, PIMS, Islamabad. SUBJECTS AND METHODS: Records of 171 consecutive singleton breech deliveries > or = 28 weeks gestation were retrieved and stratified into 3 groups by gestational age, 28-33 completed weeks, 34-36 completed weeks and beyond 37 weeks. The main outcome measures were the incidence of perinatal morbidity and mortality of the three gestation groups in relation to the mode of delivery. RESULTS: Of 171 cases, 20 women (11.7%) delivered at 28 to 33 weeks, 17 of whom were delivered vaginally. The main birth weight (MBW) was 1.4 kg. Perinatal deaths occurred in 15 cases, these included 4 intrauterine deaths (IUDs) and 6 infants with congenital anomalies. Thirty three women (19.3%) delivered at 34 to 36 weeks gestation, of whom 17 (51.5%) delivered vaginally. The MBW was 2 kg with 6 perinatal deaths (one with cephalhaematoma and 4 IUD and 2 congenital anomalies). Sixteen women (48.5%) had caesarean sections. The MBW was 2.2 kg with only 2 perinatal deaths due to IUD and congenital pneumonia. One hundred and eighteen (69%) women delivered at term. The MBW of the 45 (38.1%) vaginally delivered infants was 2.2 kg with 9 perinatal deaths (1 IUD and 7 congenital anomalies). Seventy three (61.9%) had caesarean sections, 41 emergency and 32 elective. The MBW was 2.9 kg with 4 perinatal deaths (1 IUD, 2 congenital anomalies and 1 sepsis). CONCLUSION: There was significant association of breech presentation with congenital anomalies. After the exclusion of congenital anomalies, greater recourse to caesarean section beyond 34 weeks seems to confer a survival advantage to the new born infant. PMID- 12553678 TI - Thalidomide prevents donor corneal graft neovascularization in an alkali burn model of corneal angiogenesis. AB - OBJECTIVE: Thalidomide is a potent inhibitor of angiogenesis. We evaluated the effects of Thalidomide on corneal angiogenesis and on tissue survival of grafts in rabbit eyes with pre-existing neovascularization secondary to alkali burn. METHODS: Sixteen rabbits received alkali burns to one cornea. One month post injury, assessments of corneal neovascularization were performed followed by corneal transplantation. Four rabbits received oral Thalidomide and ten got placebo (powdered sugar) for thirty days. Total corneal neovascularization (NV), clock hours (CH) involved in (NV), longest (NV) pedicle length (NVP) and the duration of time required for NV to develop were assessed. RESULTS: Thalidomide significantly decreased the total neovascularization (p<0.0072), the number of (CH) involved (p<0.0002) and the longest (NVP) length (p<0.0001). There was also a significant delay in the earliest development of NV in the test group (p<0.0064). The test group retained corneal clarity significantly longer than the control group (p<0.0008). CONCLUSION: Thalidomide is an effective inhibitor of corneal angiogenesis and prolongs graft survival as measured by graft clarity in donor corneas in eyes with previous neovascularization secondary to alkali injury. CLINICAL RELEVANCE: Thalidomide may be used as a modulator of corneal angiogenesis to prolong graft survival in eyes with pre-existing corneal neovascularization. PMID- 12553679 TI - Long-term outcomes of acute myeloid leukemia in adults in Pakistan. AB - OBJECTIVE: To describe the long-term outcomes of Acute Myeloid Leukemia (AML) and to study clinicopathological features at presentation, morphological subtypes and remission rates. METHODS: Demographic information, response to therapy and survival of patients (>14 years of age) admitted between January 1988 to August 1996 with acute myeloid leukaemia was retrieved and analysed. RESULTS: Seventy four patients were admitted with a diagnosis of AML during the study period. There were 43 males and 31 females. Age ranged between 15 and 70 years with a mean age of 38 years. The most common presenting feature was fever (67.5%) and the morphological subtype according to French-American-British Group (FAB) criteria was M4. Fifty-five patients received treatment and were evaluable for response and outcomes. Thirty-six (65.4%) patients had complete remission. Sixteen (29.1%) died during the first 28 days after starting induction chemotherapy. The median survival was 11 months. Six (11%) patients (4 females, 2 males) are surviving beyond 4 years (long-term survivors). CONCLUSION: Our study suggests that the long-term outcomes of adults with AML are comparable to what has been reported in the literature for patients who do not receive bone marrow transplants. PMID- 12553680 TI - Respiratory arrest in a child after flushing of pancuronium from the deadspace of intravenous cannula. PMID- 12553681 TI - Ameloblastoma--management and review of literature. PMID- 12553682 TI - View box case--10. Aspergilloma. PMID- 12553683 TI - Comparative analysis of knowledge, attitudes and perceptions about induced abortions among medical and non-medical students of Karachi. AB - OBJECTIVE: To determine the knowledge, attitudes and perceptions regarding induced abortions among university students of Karachi and compare these aspects among medical and non-medical students. STUDY DESIGN: Cross sectional opinion survey. METHODS: The study is a cross sectional opinion survey of medical and non medical students from four educational institutions of Karachi using a pretested questionnaire. RESULTS: Of the 381 students interviewed, 201 were medical and 180 were non-medical. More medical students were aware of the correct definition of induced abortion. An encouraging finding was the awareness of our study population about the only legally acceptable circumstance for induced abortion in Pakistan, that is, if conceptus is a risk to maternal health (p<0.001). More medical students cited complications that were technical like uterine perforation (p<0.001), cervical trauma (p<0.001), DIC (p<0.001), and sepsis (p<0.001). More medical students thought induced abortion to be a health issue (p=0.001) as compared to non-medical students. Most of the medical students felt the need to modify Pakistan's law regarding abortion (p=0.008) especially for fetal anomalies (13.9%) and rape (7%). CONCLUSION: Though medical students were more knowledgeable about induced abortions as compared to the non-medical population, there is a need to improve awareness regarding induced and unsafe abortions and their consequences. PMID- 12553684 TI - Immune responses against HIV: what we want and what we get. PMID- 12553685 TI - Mechanisms of loss of HIV-1-specific T-cell responses. AB - Virus-specific T-cell responses, important for the control of HIV-1 infection, are seen in HIV-1-infected subjects in the early stages of infection. A progressive variable decline in HIV-1-specific CD4 and CD8 T-cell responses during the course of the infection is not reversed by the administration of potent antiretroviral drugs. The mechanisms responsible for this HIV-1-inflicted loss are complex, involving processes ranging from T-cell ontogeny to the final stages of antigen presentation and T-cell differentiation. HIV-1-specific CD4 and CD8 T cells are present in most patients, but have been rendered anergic, either directly by HIV-1 or indirectly by clonal inactivation, exhaustion and/or suppression. The absence of functional HIV-1-specific T cells in chronic infection, even after the initiation of antiretroviral therapy, indicates that additional immunomodulatory therapy is required. To induce or maintain such cellular responses in an immunosuppressed environment of chronic infection is proving difficult to achieve. The induction of virus-specific CD4 T-cell and, subsequently, CD8 T-cell responses may require different novel approaches based on an appreciation of the complex mechanisms involved in the loss of these responses. PMID- 12553686 TI - HIV-1 and AIDS: what are protective immune responses? AB - HIV-1 is the leading cause of death in sub-Saharan Africa, responsible for one in five deaths in the region. Although potent antiretroviral therapy has had a huge impact on HIV-associated morbidity and mortality in economically advantaged countries, it is beyond the reach of most infected people in the world. The development of an effective HIV vaccine would be a huge step towards stopping the pandemic, but an important precondition for such a vaccine is that it must induce a host immune response that can protect the host from HIV acquisition or disease progression. This article reviews the evidence that protective host immune responses do exist, either in highly exposed, persistently seronegative (HEPS) subjects or in HIV-1-infected long-term non-progressors (LTNPs), as well as efforts to reproduce putative protective immunity in animal vaccine models. HIV-1 specific cellular responses are a key to viral control in infected subjects, but generally fail in the long term. This suggests that the goal of a preventive HIV 1 vaccine should be sterile immunity, rather than improved virus control after infection. Achieving this goal will at least require the induction of HIV-1 specific cellular immune responses at the site of initial viral contact (generally the genital tract), perhaps in combination with HIV-1-specific neutralising antibody. PMID- 12553687 TI - Improving HIV-specific immune responses in HIV-infected patients. AB - Deficiencies in potentially highly protective HIV-1-specific immune responses have led to interventions with immunotherapeutic strategies such as post-exposure vaccination. The application of exogenous antigen to the HIV-infected individual by therapeutic vaccination might be expected to induce renewed virus-specific effector T-cell and neutralising-antibody responses. However, the nature of HIV-1 immunopathogenicity precludes this approach: high levels of viral turnover, persistent expression and presentation of HIV-1 antigen to the immune system, T cell hyperactivation and exhaustion and clonal T-cell anergy all successfully counter any effect of exogenous antigen. Even with the use of highly active antiretroviral therapy (HAART), when HIV-1 activity is profoundly restricted, the induction of immune responses remains problematic. Different vaccine strategies are currently being tested, including a whole killed virus preparation (Remune), a yeast virus-like particle (p24 VLP), whole antigen preparations (VaxSyn), canarypox immunogens (ALVAC) and DNA plasmids. Therapeutic vaccine strategies currently in the earliest stages of development include adenovirus vectors and a topical DNA preparation, DermaVir. PMID- 12553688 TI - Immune restoration disease in HIV patients: aberrant immune responses after antiretroviral therapy. AB - Suppression of HIV replication by antiretroviral therapy has various effects on the immune system of HIV-infected patients. Whereas protective pathogen-specific immune responses are restored in some patients, others have persistent immunodeficiency or produce immunopathological responses against opportunistic pathogens that cause immune restoration disease (IRD). Significant morbidity and even mortality may result from IRDs associated with infections by mycobacteria, herpesviruses, hepatitis viruses or the JC virus. Preliminary evidence from our studies suggests that immune responses in patients with IRD may be dysregulated and affected by immunogenetic factors. Thus, herpesvirus IRDs were associated with increased plasma levels of bioavailable interleukin-6 and soluble CD30, while mycobacterial and herpesvirus IRDs were associated with a major histocompatibility complex gene haplotype and/or alleles of particular cytokine genes. A greater understanding of pathogenic mechanisms and the immunogenetics of IRDs may lead to improved preventive and management strategies. PMID- 12553689 TI - Rumen fermentation and nitrogen balance of lambs fed diets containing plant extracts rich in tannins and saponins, and associated emissions of nitrogen and methane. AB - Tannins were added to experimental diets at levels of 1 and 2 g/kg DM (hydrolysable tannins; Castanea sativa wood extract) and saponins at 2 and 30 mg/kg DM (sarsaponin; Yucca schidigera extract). These levels were far below thresholds expected to be adverse in ruminants. Effects were measured in lambs by comparison with unsupplemented control diets calculated to be either deficient (10%) or adequate in protein. The diets consisted of hay, concentrate (1:1) and extra wheat starch with increasing body weight. Ruminal pH, VFA concentration, protozoa count and apparent digestibilities of organic matter and fibre did not differ among treatments. The low tannin dose significantly decreased bacteria count compared to the high saponin dose. Saponin supplementation and the high tannin dose showed some potential to reduce ruminal ammonia concentration. This was associated with weak trends towards lower urine N excretion (only tannins) and ammonia emission from manure. Methane release was increased by the low tannin dose compared to the unsupplemented control. Diet effects on heat production were not systematic. In conclusion, the extracts rich in tannins or saponins gave only slight indications for either increased body nitrogen retention or reduced nitrogen emission. However, effects might have been larger with more pronounced dietary protein deficit. PMID- 12553690 TI - Influence of various feeding conditions, the migrating myoelectric complex and cholinergic drugs on antral slow waves in sheep. AB - The presented study was designed to elucidate whether the cholinergic mechanisms control ovine antral slow waves in various physiological conditions, including feeding and various phases of migrating myoelectric complex (MMC). The investigations were carried out on six adult sheep of Polish Merino breed with seven bipolar electrodes surgically implanted onto the antral and small intestinal wall. In the course of chronic experiments, the myoelectric activity was recorded from these regions using the multichannel electroencephalograph. Experiments were performed on 48 h fasted and non-fasted animals. During some of these experiments, sheep were fed with standard fodder. During control experiments 0.15 M NaCl was slowly administered i.v. through the indwelling catheter and during other experiment, hexamethonium bromide (2.0 and 5.0 mg/kg). atropine sulfate (0.02; 0.1; 0.5 and 1.5 mg/kg) and pirenzepine dihydrochloride (0.02; 0.5 and 2.0 mg/kg) were administered i.v. during phase 1-2a or 2b MMC. The drugs were also given in combinations. The recordings were analysed and the antral slow wave amplitudes and frequencies were calculated. Unlike the slow wave amplitude, either feeding or the anticholinergic drugs significantly increased slow wave frequency, especially when the given procedure was started during phase 2b MMC. The most pronounced effects were observed after hexamethonium given alone or in combinations. Thus, the cholinergic system modulates antral slow wave frequency in sheep. PMID- 12553691 TI - Amino acid profile of escaped feed protein after rumen incubation and their intestinal digestibility. AB - The crude protein content and amino acid profile of seven feedstuffs (linseed meal, maize gluten meal, rapeseed meal, rapeseed meal protected, soybean meal, fullfat soybean extruded and sunflower meal) were determined before and after ruminal incubation for 16 h in three bulls with large rumen cannulas. The intestinal disappearance of amino acids was measured using mobile bag technique. Ruminal incubation affected amino acid profile of all experimental feedstuffs. Crude protein degradation varied from 29.3% for maize gluten meal to 86.4% for rapeseed meal. A tendency towards increased disappearance was observed for glutamic acid, histidine, lysine and proline and decreased disappearance for branched-chain amino acids. The intestinal crude protein digestibility was higher than > 80%, except rapeseed meal (66.4%) and sunflower meal (77.8%). The least digestible individual amino acids were methionine and isoleucine in rapeseed meal, histidine and methionine in rapeseed meal protected and arginine in sunflower meal. In general, the lowest amino acid digestibilities were found in feedstuffs with the highest fibre content. The feedstuffs show that they have different potential for supplying of limiting amino acids. Of particular value are the feedstuffs with low crude protein degradability in the rumen and high intestinal digestibility of amino acids. PMID- 12553692 TI - Influence of a probiotic Enterococcus faecium strain on selected bacterial groups in the small intestine of growing turkey poults. AB - A feeding trial was carried out with turkey poults, which were fed a diet containing 10(10) viable probiotic E. faecium NCIB 10415 cells/kg feed. Samples of the intestinal tract were analyzed for lactate, colony forming units of total anaerobic bacteria, lactic acid bacteria, enterobacteria and enterococci. Furthermore, metabolic activity of total eubacterial, lactobacilli and enterococci was recorded in selected RNA-extracts with specific ribosomal RNA oligonucleotide probes. Animals fed the probiotic diet showed continously increasing lactate concentrations throughout the sampling period up to day 42 of life. No correlation was found for colony forming units (cfu) of lactic acid bacteria, but metabolic activity of lactobacilli showed very close relation to continously increasing lactate concentrations. Throughout the feeding trial, enterococci in the control group continously increased to a maximum of 10(4) cfu/g wet weight, but 10-fold higher enterococci cfu were generally found in the treated group. However, rRNA content as measure for metabolic activity showed a drastic decline in both groups after high metabolic activities on day 7. This study shows that E. faecium NCIB 10415 (E. faecium SF68) stimulates other lactic acid bacteria in the small intestine, especially lactobacilli. PMID- 12553693 TI - Effect of L-carnitine supplementation on utilisation of energy and protein in broiler chicken fed different dietary fat levels. AB - Effects of a supplementation of 80 mg L-carnitine per kg diet were studied in broiler chicken at two dietary levels of fat (4 and 8%) and different feeding levels (ad libitum in a growth trial, 95 and 85% of ad libitum in a balance trial). A low-carnitine basal diet adequate in amino acid concentration was used. In the growth trial, each diet was fed to 9 groups of 10 birds each for 16 days from day 5 of live onwards. Growth and feed intake were determined. At the end of the trial, birds were killed and homogenised for subsequent empty body analysis. Accretion of protein and energy was determined using a representative blank group killed at the beginning of the trial. In the balance trial, 8 individual birds were used per treatment. Birds were offered the feed at approximately 85 and 95% of ad libitum intake, which was determined with separate birds for both fat levels. Excreta were quantitatively collected three times daily for 8 consecutive days beginning on day 17 individually for each bird. Supplemented L-carnitine did not significantly affect any response criterion. However, growth and feed conversion tended to be improved by about 5% in the carnitine supplemented diets when fed ad libitum. An interaction between carnitine and fat level occurred with regard to feed conversion, indicating that carnitine had a positive effect at the high fat level, but not at the low fat level. L-carnitine did not positively affect the metabolisability of energy (ME/GE) and the efficiency of energy utilisation (RE/GE or RE/ME). Similarly, no significant carnitine effect was determined with regard to N accretion and the efficiency of utilisation of dietary protein in both trials. It is concluded that endogenous carnitine synthesis is not the limiting factor for energy utilisation in broiler chicken, even at high dietary fat concentration. Occasionally reported positive effects of supplemental carnitine were likewise caused by reasons other than improved energy or protein utilisation. Further studies on amino acid utilisation and catabolism should consider marginal amino acid supply. PMID- 12553694 TI - Influence of dietary protein level and the amino acids methionine and lysine on leather properties of blue fox (Alopex lagopus) pelts. AB - The influence of dietary protein, methionine, and lysine on leather quality in blue fox pelts was studied. The pelt material originated from animals in two consecutive feeding trials (Exp. 1 and Exp. 2) with three protein levels: conventional, slightly lowered, and very low. The two lowest protein diets were fed as such or as supplemented with methionine or with lysine (lysine only in Exp. 2). The following physical leather properties were measured: breaking load (BRL), tensile strength (TEN), relative elongation at break (PEB), straining of skins at pelting, and shrinkage at dressing. A decline in the dietary protein content reduced BRL and, hence, leather firmness, and increased straining and the corresponding shrinking in Exp. 1. The supplemented methionine tended to improve leather strength and elasticity by increasing TEN and PEB in Exp. 1, whereas lysine elicited no response. Methionine supplementation at the slightly lowered protein level increased BRL in both experiments by almost 10% as compared with the respective non-supplemented diet. We conclude that with high protein quality diets, a level of 200 g/kg DM (as digestible protein) appears to be adequate for producing pelts with firm, elastic leather, provided that an adequate amount of methionine is included in the diet. PMID- 12553695 TI - Histochemistry is a methodological step that brings morphology to a qualitatively different level. PMID- 12553696 TI - Collagen type IV in epithelial tumours of colon. AB - Collagen type IV in the lamina propria mucosae is one of the main components of the basement membrane of normal and transitional colon mucosa. The aim of the present study was to assess the use of anti-collagen type IV antibodies in the evaluation of biological activity of epithelial tumours of the colon. Formalin fixed and paraffin-embedded specimens of polyps and carcinomas of the colon from 14 patients were analyzed. In transitional mucosa around epithelial tumours, only minor deformities of the evenly thick collagen type IV-containing basement membranes were found. This pattern was different in polyps where collagen type IV positive basement membrane components extended between basolateral membranes of epithelial cells. Local changes of collagen type IV positivity in basement membranes of polyps were observed. Positivity of epithelial basement membranes disappeared in adenocarcinomas but there was an increased positivity in fibrillar components of stroma. Basement membranes of microvessels in lamina propria mucosae were also positive for collagen type IV. Similar observations were made in the stroma of polyps. Our results indicate that loss of collagen type IV in basement membranes of adenocarcinomas is related to loss of differentiation and the malignant potential of epithelial tumours of colon. PMID- 12553697 TI - Expression of proliferation and apoptotic markers in human placenta during pregnancy. AB - Trophoblast has unique properties in relation to its wide range of metabolic, endocrine and angiogenic functions. Trophoblastic cells invade endometrium and adjacent myometrium in a way that is imitated by malignant tumours. The aim of the present study was to analyse the expression of markers of proliferation and apoptosis in trophoblastic cells in normal human placenta during pregnancy. A total of 22 placentas, 12 of which were obtained from curettage and induced legal abortion and 10 placentas obtained from normal deliveries or caesarean sections were included in this study. Proliferation markers were strongly expressed in cytotrophoblast in early stages of gestation. In late term placentas, a distinct decrease in expression of these markers was observed. Syncytiotrophoblast was negative for proliferation markers in all placentas. Positive immunostaining for bcl-2, an anti-apoptotic marker, was observed only in syncytiotrophoblastic cells in first-trimester but also in third-trimester placentas. Cytotrophoblast and stromal mesenchymal cells of chorionic villi were negative for bcl-2. Expression of bcl-2 protein in syncytiotrophoblast may be one of the major factors preventing these structures from early cell death, which is indispensable for the maintenance of physiological pregnancy. PMID- 12553698 TI - Changes in sialic acid expression in the lung during intrauterine development of the human fetus. AB - Sialic acid is a component of glycoproteins that influences enzymatic and receptor functions of cells. During proliferation and differentiation of tissues, sialic acid can serve as a recognition determinant in intercellular communication and interactions of cells with the extracellular matrix. In the present study, sialic acid expression in relation to developmental maturity of the lung has been studied. We analyzed 12 necroptic lung specimens from foetuses of different gestational ages from the 15th week to the neonate. Sections were stained histochemically using 3 lectins specific for sialic acid: Tritrichomonas mobilensis lectin (TML), specific for sialic acid without linkage preference, Sambucus nigra agglutinin (SNA), specific for alpha2,6-linked sialic acid, and Maackia amurensis leucoagglutinin (MAL), specific for alpha2,3-linked sialic acid. MAL positivity dominated over SNA positivity showing prevalence of alpha2,3 linked sialic acids to be homogeneously distributed in the lung at the canalicular stage of development. In more mature lungs, well-differentiated bronchial epithelium showed strong sialic acid expression of both linkages. Sialic acid with alpha2,6 linkage dominated in vascular endothelium. Our results showed a slight decrease in sialic acid expression in lungs with gestational age to a relative minimum before birth. Lectin staining of mature lung tissue showed intense sialic acid expression in alveolar epithelial type II cells. Changes in expression of specific sialic acids during differentiation of the lungs may be useful as marker of the degree of maturity of the foetus. PMID- 12553699 TI - Sialic acid expression in autoimmune thyroiditis. AB - Autoimmune diseases of the thyroid gland are among the most frequent endocrine disorders. The present study analyzes expression patterns of sialic acids in these diseases. Three lectins specific for sialic acids were used for the histochemical analysis of surgical specimens of the thyroid gland: Tritrichomonas mobilensis lectin that stains all types of sialic acids, Maackia amurensis leukoagglutinin that stains sialic acids with alpha2,3 linkage and Sambucus nigra agglutinin that stains sialic acids with alpha2,6 linkage. In autoimmune thyroiditis, there was a significant increase in sialic acid expression in epithelial cells, especially on luminal membranes of follicular cells. The alpha2,3 linkage dominated over the alpha2,6 linkage. Lymphocytes of patients with Hashimoto thyroiditis, especially in germinal centers, showed strong expression of alpha2,6-linked sialic acids on their cell membrane. Vascular endothelium was positive in all specimens. It can be concluded, that there is a significant increase in sialic acid expression in autoimmune diseases of the thyroid gland, predominantly of sialic acids with alpha2,3 linkage, whereas the sialylation pattern of lymphocytes in Hashimoto thyroiditis was also different. PMID- 12553700 TI - Adrenergic and acetylcholinesterase-positive innervation of palatine tonsils in mammals. AB - Innervation of human, feline and rabbit palatine tonsils was investigated. Adrenergic nerve components were visualized by formaldehyde-induced fluorescence of catecholamines and 5-HT, or by glyoxylic acid fluorescence, whereas acetylcholinesterase (AChE)-positive nerve structures were demonstrated by the direct thiocholine method. The largest density of adrenergic and AChE-positive nerve profiles was found in the adventitia of arterial branches in the fibrous capsula and septa, mainly in the form of periarterial nerve plexuses of different density. Fine nerve fibres lined the wall of small arteries which penetrated into extrafollicular lymphoid tissue and marginal layers of follicles. It is concluded that there are significant species-specific differences related to density, nature and topographic relations of adrenergic and AChE-positive nerve fibres in the various structural parts of palatine tonsils. PMID- 12553701 TI - Fluorescence of hematoxylin and eosin-stained histological sections of the human spleen. AB - A major problem in the morphometric evaluation of human spleen is the simple but reliable determination of the border between T-cell and B-cell dependent areas, and other structures of the spleen. It was investigated whether cryostat sections of frozen surgical specimens of the human spleen and sections of paraffin embedded specimens could be used for this purpose after being stained with haematoxylin and eosin and mounted in autofluorescence-free medium for fluorescence microscopical evaluation. Comparison was made with sections that were immunohistochemically-stained for fibronectin and collagens type II and type IV. Both in cryostat sections and paraffin sections, fluorescence was found in circumferential reticulum of periarterial lymphatic sheets, arterial terminals, arterial walls and walls of red pulp sinuses in the spleen. Evaluation was hindered by fluorescence of erythrocytes in paraffin sections but not in cryostat sections. Results were similar as those obtained with immunohistochemical fibronectin staining and are sufficient for morphometric evaluation or orientation in the tissue in case of neoplasia. PMID- 12553702 TI - Mast cell infiltration in the wall of varicose veins. AB - Varicose veins of the lower extremities are abnormally dilated, tortuous and elongated. The exact cause of vein dilatation has still not been established. Mast cells produce, store and release various types of vasoactive compounds (histamine, tryptase, prostaglandins, leukotrienes, and cytokines). Histamine enhances local vasopermeability and smooth muscle cell proliferation, leading to thickening of the intima. Tryptase can contribute to local vascular injury and subsequent weakness of the vascular wall causing varix formation. The aim of the present study was the comparison of mast cell infiltration in the wall of varicose and non-varicose veins. The mean mast cell density in the wall of varicose veins was 0.86 mast cell per mm2 and in healthy non-dilated vein walls, density was 1.23 mast cell per mm2. This difference was not statistically significant, therefore we could not confirm our hypothesis. Nevertheless, we suggest that mast cells could play an important role in the development of varices and the factor released by the mast cells should be further examined. PMID- 12553703 TI - Histological and histochemical analysis of embryoid bodies. AB - We examined the histological structure of embryoid bodies arising from aggregation of mouse embryonic stem (ES D3) cells after 7, 12, 18 and 26 days of in vitro culture. Morphology of originally solid embryoid bodies was affected by the process of cavitation that resulted in formation of cystic embryoid bodies and by spontaneous differentiation of the ES D3 cells. We applied in situ immunophenotyping to characterise cell populations that spontaneously differentiated inside the embryoid bodies in the various stages. The most distinct cell populations that were found inside embryoid bodies were alpha fetoprotein-positive endodermal cells and myogenic cells that expressed desmin, myogenin or smooth muscle actin. ES D3-derived endothelial cells generated during vasculogenesis inside the embryoid bodies differed from mature endothelial cells because they did not stain for von Willebrand factor. These cells also differed from endothelial cell that were generated during angiogenesis since they did not stain for the intermediate filament nestin. Our results demonstrate the usefulness of this in vitro model for studying early embryogenesis. PMID- 12553704 TI - Comparison of the TUNEL, lamin B and annexin V methods for the detection of apoptosis by flow cytometry. AB - The present study was performed in order to compare the sensitivity of 3 apoptosis-detection methods using a model cell culture after induction of apoptosis. Evaluation was performed by means of flow cytometry and light microscopy. It appeared that both TUNEL and annexin V methods are sensitive and specific and produced similar data in all measurements. The immunocytochemical detection of lamin B was less reliable than the other methods. PMID- 12553705 TI - Computer-assisted quantitative analysis of immunofluorescence staining of the extracellular matrix in rat dorsal and ventral spinal roots. AB - The endoneurial extracellular matrix (ECM) is produced by Schwann cells and fibroblasts under the control of axons. Dorsal and ventral spinal roots contain different types of axons, but information is not available on differences in the composition of their ECM. A comparison was made of the intensity of immunofluorescence staining of chondroitin sulfate proteoglycan, fibronectin, tenascin and thrombospondin in the endoneurial ECM of rat dorsal and ventral spinal roots. Sections of dorsal and ventral roots were incubated simultaneously for indirect immunofluorescence detection of the epitopes studied. Brightness of immunofluorescence staining was assessed by computer-assisted image analysis using interactive segmentation of digitized images to select areas to be analyzed. Our results revealed quantitative differences in the composition of endoneurial ECM of spinal dorsal and ventral roots, probably due to the presence of different types of axons. The ECM composition of the endoneurium in dorsal and ventral roots may be related with the creation of extrinsic conditions that support differential regeneration of afferent and motor axons after injury. PMID- 12553706 TI - The reaction of the subependymal layer of lateral brain ventricles to striatal ibotenic acid lesions in a long-term study. AB - The proliferative activity in the subependymal layer of lateral brain ventricles in adulthood is known. We were interested in the reaction of this layer to ibotenic acid lesions, which simulate neurodegenerative processes in Huntington's disease. Animals with a unilateral ibotenic acid lesion were compared with sham lesioned animals and control animals with intact brains at 5 and 13 weeks after surgery. Five weeks after surgery, increased proliferation was found in most GFAP positive astrocytes and to a lesser extent in CNPase-positive oligodendrocytes in comparison with controls. Interestingly, a slight increase in proliferation was found as well in the contralateral non-lesioned hemispheres. Moderate elevation of cell proliferation was found after induction of sham-lesions as well. The intensity of the reaction in the subependymal layer decreased in the following 8 weeks. Only a few scattered cells that originated from the subependymal layer had migrated over a short distance to adjacent brain tissue. We conclude that the reaction of the subependymal layer is (a) non-specific, as it is a response to any type of lesion, and (b) slowly decreases in time. PMID- 12553707 TI - Localization of c-Fos protein in the rat spinal cord after carrageenan treatment. AB - We have characterized segmental and laminar distribution patterns of Fos immunopositive (Fos-IP) neurons in spinal cord segments L3-L6 after carrageenan treatment. A large number of Fos-IP neurons was found in the medial region of the ipsilateral dorsal horn laminae I-II at 4 and 6 h postinjection (pi). At one day pi, the number of Fos-IP neurons was decreased significantly, which correlated with suppression of inflammation in the affected hind paw. Bilaterally, Fos-IP neurons reappeared in the L3-L6 spinal cord segments at 3-4 days pi, mainly in the deep laminae LV-LVI. However, signs of inflammation had distinctly attenuated. Our data indicate a biphasic trend in Fos-IP in this experimental model of inflammation. PMID- 12553708 TI - Gap junction remodelling is involved in the susceptibility of diabetic rats to hypokalemia-induced ventricular fibrillation. AB - The objective of the present study was to examine the susceptibility of diabetic rats with cardiomyopathy to hypokalemia-induced ventricular fibrillation and to localize gap junction protein connexin-43 as well as subcellular changes that may be involved in the development of severe arrhythmia. Our results showed a significantly higher incidence of sustained ventricular fibrillation in diabetic hearts as compared with control hearts, 80% vs 20%, respectively. Diabetic cardiomyopathy itself was accompanied by a distinct decrease in connexin-43 immunopositive gap junctions. Moreover, interstitial fibrosis and subcellular alterations to various degrees were observed in diabetic hearts, and a further deterioration of the ultrastructure and impairment of intercellular junctions, and a stronger local decrease in connexin-43 levels due to hypokalemia were found. These changes were heterogeneously distributed throughout the myocardium and occurred earlier and were more pronounced in diabetic hearts than control hearts. In conclusion, our results indicate that diabetic cardiomyopathy is associated with down-regulation of gap junction proteins and may account for the higher vulnerability of diabetic rats to ventricular fibrillation in combination with impairment of intercellular communication due to hypokalemia. PMID- 12553709 TI - Histochemical and ultrastructural characterisation of an arrhythmogenic substrate in ischemic pig heart. AB - The aim of the present study was to reveal by enzyme histochemistry and ultrastructural examination the possible anatomic substrate that may be the cause of high susceptibility of the pig heart to ischemia and/or reperfusion-induced severe arrhythmias. The heart of landrace pigs was subjected to 90 min of left coronary occlusion followed by 30 min reperfusion, whereby both conditions elicited arrhythmias and often even ventricular fibrillation. We found for the first time, besides common contractile cardiomyocytes, Purkinje fibers, and "transitional cells" in mid-myocardium. Transitional cells likely correspond to the recently described M cells. Importantly, these cells and Purkinje fibers exhibited reversible ischemia-related subcellular alterations, whereas the majority of contractile cardiomyocytes were irreversibly injured in the area of infarction. In correlation with these findings, glycogen-dependent phosphorylase activity was abolished, whereas it was still persistent in Purkinje fibers and small islands of contractile cardiomyocytes. Moreover, a distinct heterogeneity in the activity of all enzymes selected and subcellular alterations within a border zone were observed. These results suggest that particularly the preserved viability of specialized conducting cells spanning the ventricular wall may account for electrical disturbances that consequently contribute to increased susceptibility of the pig heart to ischemia- and reperfusion-induced severe arrhythmias. PMID- 12553710 TI - Fibre type composition of soleus and extensor digitorum longus muscles in normal female inbred Lewis rats. AB - We have analysed the fibre type composition of soleus and extensor digitorum longus (EDL) muscles of normal female 4-6-month-old inbred Lewis rats. This rat strain is used in our ongoing study of the effects of thyroid hormone on myosin heavy chain (MyHC) isoform expression. On the basis of the mATPase reaction, soleus muscles contained 96.1 +/- 2.9% of type 1 fibres supplemented by 2A fibres. EDL muscles contained type 1 (5.5 +/- 1.0%), type 2A (18.8 +/- 1.7%) and type 2B (75.7 +/- 2.2%) fibres. Immunohistochemical analysis and SDS gel electrophoresis confirmed that most fibres in the soleus muscle expressed the type 1 (slow) MyHC isoform and that only a small proportion of fibres expressed the fast 2a MyHC isoform. Immunohistochemical analysis and SDS gel electrophoresis demonstrated that almost half of the 2B fibres of EDL muscles expressed the 2x/d MyHC isoform. In both muscle types, many fibres expressed more than one MyHC isoform. The content of slow fibres in the soleus muscle of female inbred Lewis rats was slightly higher than that reported for Wistar rats, but was considerably higher than that of Sprague-Dawley rats, whereas substantial differences were not found in the proportion of slow and fast fibre types in EDL muscles in these strains. PMID- 12553711 TI - Apical ouabain-sensitive and ouabain-insensitive ATPases in rat colonic epithelium. AB - Active resorption of potassium ions in the colon is mediated by ouabain-sensitive and ouabain-insensitive H,K-ATPases localized in the apical membrane of colonic enterocytes (colonocytes). The present study was performed to investigate distribution patterns of apical ATPases using catalytic histochemistry and immunohistochemistry. Activity of the ATPases was localized both at the apical and basolateral regions of these cells. The basolateral activity was almost completely inhibited by ouabain, but apical ATPase activity was only partially inhibited by ouabain. Ultracytochemically, activity was localized at the cytoplasmic side of the apical and basolateral membrane and thus we assume that the activity represents isoforms of apical H,K-ATPases and basolateral Na,K ATPase. With the use of a polyclonal antibody raised against H,K-ATPase alpha subunit, we demonstrated immunostaining only in the apical region of colonic enterocytes whereas positive staining was not observed at the basolateral membrane. On the other hand, when antibodies against alpha1-subunit Na,K-ATPase were applied, immunostaining was localized only in the basolateral membrane domain. Therefore, we conclude that ATPases as demonstrated histochemically in the present study were identified immunohistochemically as colonic alpha-subunit H,K-ATPase (in the apical cell membrane of colonocytes along the entire length of crypts) and as alpha1-subunit Na,K-ATPase (in the basolateral membrane). PMID- 12553712 TI - The pyridoindole antioxidant stobadine attenuates histochemical changes in kidney of streptozotocin-induced diabetic rats. AB - The aim of the present study was to investigate the effects of dietary supplementation with the pyridoindole antioxidant stobadine on histochemical parameters in kidney of streptozotocin-induced diabetic rats. Diabetic male Wistar rats were fed a standard diet or a diet supplemented with stobadine (0.05% w/w) for 24 weeks. The diabetic state was characterized by significantly elevated plasma levels of glucose and glycated hemoglobin, severe reduction of total body weight and relatively enlarged kidneys. Kidney alkaline phosphatase activity was not changed by diabetes. Activity of 5'-nucleotidase, K(+)-dependent p nitrophenylphosphatase, ATPase and mitochondrial succinic dehydrogenase were markedly decreased in kidneys of diabetic rats. In contrast, activity of beta hydroxybutyrate dehydrogenase was moderately increased in kidney of diabetic rats as compared to controls. Long-term treatment of diabetic animals with stobadine attenuated histochemical changes in kidney tissue. PMID- 12553713 TI - Histochemical demonstration of the heterogeneity of the epithelium of proximal tubules in the chick mesonephros. AB - Proximal tubules (PT) in 7-10-day old chick mesonephros were examined histochemically to evaluate their structural and functional properties related to the absorption capacity of the epithelium and its possible alterations leading to cystically dilated tubules (CDT). Alkaline phosphatase activity at the apical cell membrane was demonstrated in varying intensities in large PT. A similar heterogeneity was also detected in the expression of proteoglycans (Lewis(x) antigen) localized in the apical part of the epithelial cell membrane but not in the basolateral membrane parts (beta-catenin, Na(+),K(+)-ATPase). In analogy, the ability to accumulate trypan blue was found in the same cell population. We hypothesize that epithelial cells in proximal tubules of nephrons with a defective apical cell membrane cause reduced fluid absorption and subsequent overfilling and dilation of the tubules. PMID- 12553714 TI - Histochemical localization of NADPH-diaphorase in neurons of the pheasant ileum. AB - Localization patterns of NADPH-diaphorase-positive neurons in the pheasant ileum were investigated using an enzyme histochemical method. NADPH-diaphorase activity in the pheasant ileum was demonstrated in neuronal cells bodies and nerve fibres. The NADPH-diaphorase-positive nerve cells showed a polygonal shape and were present solitary or arranged in groups in the submucosal and muscular layers. Nerve fibres penetrated the wall of the ileum at its serosal surface, frequently in the vicinity of ileal arterial branches. They were abundantly present in muscular and submucosal layers of the ileum forming thicker nerves. Some nerve fibres traversed the submucosa into the lamina propria mucosae to form dense nerve plexuses. Fine nerve fibres were found to penetrate into intestinal villi encompassing the crypts underneath the epithelium. We conclude that the pheasant ileum is characterized by abundance of NADPH-diaphorase-positive nerve structures which may play a significant functional role in the small intestine of the pheasant. PMID- 12553715 TI - NADPH-diaphorase activity in the spinal cord after ischemic injury and the effects of pretreatment with Ginkgo biloba extract (EGb 761). AB - Histochemical analysis of NADPH-diaphorase (NADPH-d) activity was performed on segments of the lumbar spinal cord in rabbit after 7 days pretreatment with the Ginkgo biloba extract Tanakan, and 30 min of ischemia followed by 24 h of reperfusion. In sections of the L5 segment of the spinal cord of untreated controls, NADPH-d-positive neurons were identified in the dorsal horns, in the pericentral region and occasionally in the ventral horns. The rabbits were completely paraplegic after 30 min of ischemia and 24 h of reperfusion. High NADPH-d activity was found in the wall of blood vessels in sections of the L5 segment and the numbers of NADPH-d-positive neurons in all sites was moderately elevated. After 7 days of Tanakan pretreatment, 30 min of ischemia and 24 h of reperfusion, the animals did not show paraplegia. Only a light tremor of the hind limbs was observed. NADPH-d activity in blood vessels and neurons was similar to that in controls. In the dorsal horns, NADPH-d positivity in neurons and fibres was increased. Our results indicate that Tanakan can scavenge free radicals produced during ischemia/reperfusion and may reduce reperfusion damage. PMID- 12553716 TI - Demonstration of intermediate filaments in sheep ovary. AB - Distribution patterns of intermediate filaments, vimentin, smooth muscle actin, and desmin were studied in the ovine ovary using an immunohistochemical method. Vimentin was consistently expressed in follicular cells of primary, secondary, and antral follicles. Stromal cells of cortex and medulla and those surrounding the corpus luteum were immunostained with the anti-vimentin antibody. Endothelial cells lining blood vessels showed strong anti-vimentin positivity. Smooth muscle cells positive for smooth muscle actin were incorporated in the cortical region in the theca interna and externa and formed incomplete spheres around large antral follicles. In atretic follicles, the presence of positive smooth muscle cells inside follicular spaces were related with capillaries. Corpora lutea were surrounded by a layer of positive smooth muscle cells. In the hilus, smooth muscle actin was localized in the wall of blood vessels. In cortical regions, desmin-positive cells were randomly distributed and occasionally formed ill defined clusters around tertiary follicles, but not around secondary follicles, whereas desmin was expressed in the medullary region in blood vessels. The distribution pattern of intermediate filaments in sheep ovary suggests a possible role of smooth muscle cells in the mechanism of ovulation. PMID- 12553717 TI - T lymphocyte subpopulations and B lymphocyte cells in caecum and spleen of chicks infected with Salmonella enteritidis. AB - The effect of low and high doses of Salmonella enteritidis PT4 (SE) on immunocompetent cells in caecum and spleen of one-day-old chicks was investigated. Subsets of T lymphocytes positive for CD3, CD4, CD8 and B lymphocytes (Bu1b-positive cells) were counted in the caecum after immunohistochemical staining and the relative percentage of these cells in the spleen was analysed using a FACScan cytometer on days 7, 10, 14, 21, and 27 post inoculation (pi). In the low dose group, the number of CD3+ and CD4+ cells in the caecum had significantly increased at day 10 pi. Both CD8+ and Bu1b+ cells were significantly higher on day 14 pi in this group. In the high-dose group, the number of CD4+ cells had significantly increased at day 7 pi. CD3+, CD8+, and Bu1b+ cells showed prolonged proliferation at days 7 up to 21 pi. Splenic lymphocytes demonstrated significant changes only in the high dose group. The percentage of splenic CD4+ cells was decreased at day 7 pi. A decrease in CD3+ and CD8+ cells was found at day 14 pi in this group. PMID- 12553718 TI - Localization and measurement of extracellular plant galactosidases. AB - A simple, rapid and sensitive procedure for the identification and determination of plant extracellular alpha-galactosidase and beta-galactosidase is described using callus cultures and seedlings from tomato. Synthetic substrates (1-naphthyl and p-nitrophenyl-alpha-D- and beta-D-galactopyranosides) were used for the identification and determination of intracellular and extracellular activity of alpha-galactosidase and beta-galactosidase, respectively. Many iminosugars or azasugars are strong glycosidase inhibitors and some of them show promising chemotherapeutic effects against viral diseases, and are potentially antidiabetic agents, as well as antitumor agents. These facts initiated our interest in a rapid and sensitive assay to determine activity of alpha-galactosidase and beta galactosidase in plant tissues. The results presented here show the potential of the assay of the activity of intracellular and extracellular galactosidases of plant origin in inhibitory and/or biotechnological studies. PMID- 12553719 TI - Processing and activation of lysosomal proteinases. AB - Lysosomal proteinases are translated as preproenzymes, transferred through the Golgi apparatus as proenzymes, and localized in lysosomes as the mature enzymes. Pulse-chase analyses and the immunoisolation of proenzymes or recombinant proenzymes are useful tools for analyzing this process, but the processing proteinases that participate in this pathway are largely unknown. Recently, we developed a new method for analyzing processing proteinases using Bafilomycin A1 and proteinase inhibitors. Here we summarize the recent progress including our results obtained using this method. PMID- 12553720 TI - Evidence that E-box promoter elements and MyoD transcription factors play a role in the induction of cathepsin B gene expression during human myoblast differentiation. AB - HB13 human myoblasts express physiological and biochemical markers associated with myoblast differentiation in non-human cell culture model systems. During differentiation, HB13 myoblasts also demonstrate fusion-related increases in cathepsin B activity and protein levels. These increases are associated with an increase in levels of cathepsin B mRNA suggesting the involvement of transcriptional regulatory mechanisms. To examine these mechanisms human myoblasts were transfected with cathepsin B nested deletion promoter constructs within the 1.8 kb 5' promoter 1 region of the human catB gene. Transfected myoblasts that were maintained under differentiating conditions demonstrated higher promoter activity than those maintained in proliferating conditions. The highest activity was obtained with pSCB2-3 (-1279/+56 bp), a construct containing two putative upstream E-box elements. Co-transfection experiments demonstrated that MyoD and myogenin transactivate cathepsin B promoter activity. Electrophoretic mobility shift assays of nuclear extracts incubated with an oligonucleotide containing two upstream E-box elements found within the cathepsin B promoter demonstrated two band shifts. The band shifts were abolished using an oligonucleotide with mutations in both E-box elements. Moreover, the shifted bands were super-shifted and abolished when incubated with anti-myogenin and anti MyoD, respectively. Collectively, these data support myogenic transcription factor-mediated activation of cathepsin B expression during myogenesis. PMID- 12553721 TI - Transcription factor Egr-1 activates collagen expression in immortalized fibroblasts or fibrosarcoma cells. AB - Synovial fibroblasts from rheumatoid arthritis patients express elevated levels of the transcription factor Egr-1. The metabolic consequences of Egr-1 overexpression in fibroblasts are not known in detail. Therefore we searched for gene products that are differentially expressed in Egr-1(high) versus Egr-1(low) fibroblasts. Immortalized synovial fibroblasts were transfected with two different Egr-1 expression vectors. Expression of recombinant Egr-1 was confirmed by RT-PCR and immunoblots. Random arbitrarily primed PCR revealed that Egr-1 induces enhanced transcription levels of the alpha1 chain of type I collagen. Increased expression of the alpha2 (I) chain could also be observed. We found enhanced levels of type I collagen propeptide in supernatants and stronger signals of alpha2 (I) protein in extracts of the Egr-1(high) expressing clone versus controls. Additionally, Egr-1 was transiently expressed in fibrosarcoma cells. These cells showed a pronounced elevation of type I collagen (alpha1) transcripts as well. Moreover, we could demonstrate that Egr-1 induces transcription of other genes including type II collagen (alpha1) and plateled derived growth factor beta1. These data suggest that upregulation of Egr-1 might contribute tofibrosis observed in rheumatoid arthritis synovium by activation of genes encoding the alpha1 and alpha2 chains of type I collagen. PMID- 12553722 TI - The receptor-bound N-terminal ectodomain of the amyloid precursor protein is associated with membrane rafts. AB - The soluble N-terminal ectodomain of amyloid precursor protein (sAPP), resulting from alpha-secretase-mediated proteolytic processing, has been shown to function as a growth factor for epithelial cells, including keratinocytes and thyrocytes. Extracellularly applied sAPP binds to a cell surface receptor and exhibits a patchy binding pattern reminiscent of that observed for raft proteins. Here we show that (i) the receptor-bound sAPP resides in a detergent-insoluble membrane microdomain which cofractionates in density gradients with cholesterol-rich membrane rafts and caveolae; (ii) the sAPP-binding microdomains are different from caveolae; and (iii) sAPP is capable of binding to isolated rafts and inducing tyrosine phosphorylation of some raft proteins. These observations suggest that a novel type of membrane raft is involved in sAPP signaling. PMID- 12553723 TI - Evaluation of similarities in the cis/trans isomerase function of trigger factor and DnaK. AB - Two functionally redundant enzymes, trigger factor and the hsp70 chaperone DnaK, have been found to assist de novo protein folding in E coli. Trigger factor is a peripheral peptidyl prolyl cis/trans isomerase (PPIase) of the large subunit of the ribosome. In contrast, DnaK displays two catalytic features: the secondary amide peptide bond cis/trans isomerase (APIase) function supplemented by the ATPase site. APIases accelerate the cis/trans isomerization of nonprolyl peptide bonds. Both enzymes have affinity for an unfolded polypeptide chain. The diminished low temperature cell viability in the presence of trigger factor variants with impaired PPlase activity indicates that the enhancement of folding rates plays a crucial role in protein folding in vivo. For the DnaK-mediated increase in the folding yield in vitro, the minimal model for APlase catalysis involves the catalyzed partitioning of a rapidly formed folding intermediate as could be inferred from the DnaK/DnaJ/GrpE/ATP-assisted refolding of GdmCl denatured luciferase. Using three different peptide bond cis/trans isomerization assays in vitro, we could show that there is no overlapping substrate specificity of trigger factor and DnaK. We propose that only if trigger factor recruits supplementing molecules is it capable of exhibiting functional complementarity with DnaK in protein folding. PMID- 12553724 TI - The chloroplast import receptor Toc34 functions as preprotein-regulated GTPase. AB - Toc34 is a protein of the chloroplast outer envelope membrane that acts as receptor for preproteins containing a transit sequence. The recognition of preproteins by Toc34 is regulated by GTP binding and phosphorylation. The phosphorylation site of Toc34 is located at serine 113, close to the postulated triphosphate binding site. This can explain the down-regulation of Toc34 by phosphorylation, resulting in the loss of GTP binding. Vice versa, GTP but not GDP binding of Toc34 influences the phosphorylation. The nucleotide specificity of Toc34 is not only determined by the classical nucleotide binding domains but by a non-typical region at the N-terminus of the protein. As a result, the GTP binding properties are unusual, since the triphosphate moiety of GTP is bound with higher affinity than the purine base. Purified Toc34 hydrolyses GTP at a low rate, which could regulate the receptor function. The rate of hydrolysis is greatly stimulated by a precursor protein. PMID- 12553725 TI - Cis-4-methylsphingosine phosphate induces apoptosis in neuroblastoma cells by opposite effects on p38 and ERK mitogen-activated protein kinases. AB - Intracellular phosphorylation of cis-4-methylsphingosine was previously shown to result in a metabolically stable compound that accumulates in Swiss 3T3 fibroblasts and mimics the mitogenic effect induced by the short-lived sphingosine metabolite, sphingosine-1-phosphate. In the present study incubation of neuroblastoma B104 cells with cis-4-methylsphingosine (10 microM) also resulted in an intracellular accumulation of its phosphorylated derivative that was, however, associated with the concentration-dependent induction of apoptosis, not observed after treatment with 10 microM of sphingosine-1-phosphate or sphingosine, respectively. In B104 cells, cis-4-methylsphingosine stimulated p38 mitogen-activated protein kinase (p38 MAPK) and simultaneously inhibited extracellular signal-regulated kinase (ERK), whereas sphingosine and sphingosine 1-phosphate only stimulated p38 MAPK without suppression of ERK. Inhibition of cis-4-methylsphingosine phosphorylation reduced both, apoptosis and concurrent regulation of mitogen-activated protein kinases (MAPKs), suggesting that the unusual accumulation of the phosphorylated sphingoid base was responsible for the biological effects. Furthermore, inhibition of p38 MAPK prevented cis-4 methylsphingosine-induced apoptosis, while suppression of the ERK pathway in the presence of sphingosine or sphingosine-1-phosphate resulted in apoptosis, indicating that the simultaneous opposite regulation of the two MAPKs was required for the induction of apoptosis. PMID- 12553726 TI - Evidence for the involvement of the unique C-tail of S100A9 in the binding of arachidonic acid to the heterocomplex S100A8/A9. AB - Protein complexes formed by S100A8 and S100A9 represent the only AA-binding capacity in the human neutrophilic cytosol and are involved in the intracellular arachidonic acid metabolism. The formation of S100A8/A9 protein complexes and the binding of calcium to the complexes are prerequisites for the specific binding of polyunsaturated fatty acids. The present study was undertaken to characterize the fatty acid binding site within the protein complex. Deletions at both termini and point mutations of different basic amino acids especially within the extended C terminal tail of human S100A9 were introduced. The S100A9 mutant proteins were then analyzed with respect to protein-protein interaction (GST pull down-assay and yeast two-hybrid system) and functional properties (arachidonic acid and calcium binding). The data give strong evidence that the unique C-tail of S100A9 containing the three consecutive histidine residues (His103-His105) represents the region to which the fatty acid carboxy-group is bound to the protein complex. The localization of the AA-binding site within the unique C-tail of S100A9 correlates with the fact that fatty acid binding has not yet been reported for other S100 proteins. PMID- 12553727 TI - The primary structure of three hemoglobin chains from the indigo snake (Drymarchon corais erebennus, Serpentes): first evidence for alphaD chains and two beta chain types in snakes. AB - The hemoglobin of the indigo snake (Drymarchon corais erebennus, Colubrinae) consists of two components, HbA and HbD, in the ratio of 1:1. They differ in both their alpha and beta chains. The amino acid sequences of both a chains (alphaA and alphaD) and one beta chain (betaI) were determined. The presence of an alphaD chain in a snake hemoglobin is described for the first time. A comparison of all snake beta chain sequences revealed the existence of two paralogous beta chain types in snakes as well, which are designated as betaI and betaII type. For the discussion of the physiological properties of Drymarchon hemoglobin, the sequences were compared with those of the human alpha and beta chains and those of the closely related water snake Liophis milians where functional data are available. Among the heme contacts, the substitution alphaD58(E7)His-->Gln is unusual but most likely without any effect. The residues responsible for the main part of the Bohr effect are the same as in mammalian hemoglobins. In each of the three globin chains only two residues at positions involved in the alpha1/beta2 interface contacts, most important for the stability and the properties of the hemoglobin molecule, are substituted with regard to human hemoglobin. On the contrary, nine, eleven, and six alpha1/beta1 contact residues are replaced in the alphaA, alphaD, betaI chains, respectively. PMID- 12553728 TI - Expression of the tissue kallikrein-kinin cascade in granulosa cells of the ovary. AB - The serine protease, tissue kininogenase (kallikrein), belongs to a unique family of enzymes that cleaves the decapeptide, kallidin, from the endogenous substrate kininogen. By analysis of genealogy patterns rat KLK gene family members have been detected in ovarian luteinizing granulosa cells of both gonadotrophin treated and non-treated control rats. Recently, we demonstrated that tissue kininogenase showed intense immunolabeling in angiogenic endothelial cells isolated from bovine mature and regressing corpora lutea. Therefore, the question to answer was whether granulosa cells associated with ovarian vascularization possess the same capacity to express the kallikrein-kinin cascade as do microvascular endothelial cells. As a first step, experiments were designed to determine the expression and visualization of tissue kininogenase (both active and pro forms) as well as kininogen and kinin receptors in granulosa cells of different developmental stages and segments of the ovarian follicle by immunoperoxidase assay, confocal fluorescent microscopy and in situ hybridization. PMID- 12553729 TI - Chondroitin sulfate proteoglycan is a potent enhancer in the processing of procathepsin L. AB - The acceleration effect of chondroitin-4-sulfate(CS-) proteoglycan on the processing of procathepsin L in vitro was investigated using enzyme purified from the culture medium of MLC cells. Procathepsin L was slightly processed even when it was incubated without CS-proteoglycan for 60 min in 50 mm acetate buffer, pH 5.5, and trace amounts of the 31 kDa mature form and 35-38 kDa intermediates of cathepsin L were formed. On the other hand, in the presence of CS-proteoglycan, procathepsin L was completely converted to the mature form within the same 60 minute time period. Moreover, Z-Phe-Arg-MCA hydrolyzing activity was increased significantly by the incubation with CS-proteoglycan, while no considerable increase in the activity was observed during the incubation without CS proteoglycan. Since the specific cathepsin L inhibitor, CLIK-195, inhibited the processing of procathepsin L accelerated by CS-proteoglycan, the trace amount of cathepsin L activity may participate in the processing. These results suggest that CS-proteoglycan may play a role in accelerating the processing of procathepsin L as an endogenous enhancer in the extracellular environment in vivo. PMID- 12553730 TI - Tyrosinase-mediated oxidation of acetaminophen to 4-acetamido-o-benzoquinone. AB - Based on its monophenolic structure and given its pharmacological and toxicological importance, the ability of tyrosinase to oxidize acetaminophen was studied for the first time. Progress curves showed a transient phase characteristic of the monophenolase activity of tyrosinase prior to attaining the steady-state. The duration of this transient phase strongly increased with the drug concentration, which would partly explain why paracetamol oxidation by tyrosinase has not been studied hitherto. The pathway is enhanced by the presence of minute amounts of L-dopa, which shortens the length of the lag period. Acetaminophen oxidation was inhibited by tropolone, a selective inhibitor of tyrosinase. The presence of the corresponding o-diphenol as intermediate was demonstrated with ascorbic acid by chemical oxidation using NaIO4 and by HPLC analysis, indicating that acetaminophen is oxidized by the monophenolase activity of tyrosinase to its corresponding o-quinone. These results contribute to our knowledge of the oxidation mechanisms of acetaminophen. PMID- 12553731 TI - Properties of baculovirus particles displaying GFP analyzed by fluorescence correlation spectroscopy. AB - Recombinant baculovirus particles displaying green fluorescent protein (GFP) fused to the major envelope glycoprotein gp64 of the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) were characterized by fluorescence correlation spectroscopy (FCS). FCS detected Brownian motion of single, intact recombinant baculovirus display particles with a diffusion coefficient (D) of (2.89 +/- 0.74) x 10(-8) cm2s(-1) and an apparent hydrodynamic radius of 83.35 +/ 21.22 nm. In the presence of sodium dodecyl sulfate (SDS), Triton X-100, and octylglucoside, the diffusion time was reduced to the 0.2 ms range (D = 7.57 x 10(-7) cm2s(-1)), showing that the fusion proteins were anchored in the viral envelope. This allowed for a calculation of the number of single gp64 fusion proteins incorporated in the viral membrane. A mean value of 3.2 fluorescent proteins per virus particle was obtained. Our results show that FCS is the method of choice for studying enveloped viruses such as a display virus with one component being GFP. PMID- 12553732 TI - The promoter context determines mutual repression or synergism between NF-kappaB and the glucocorticoid receptor. AB - While the biochemical mechanisms mediating repression of NF-kappaB activity by glucocorticoids (GCs) are relatively well studied, the role of promoter architecture for the effects of GCs on NF-kappaB remains poorly characterized. Therefore we constructed a set of synthetic promoter reporter constructs containing various numbers of GC-responsive elements (GREs) in distinct distances to NF-kappaB binding sites. TNFalpha-induced activity of a synthetic promoter controlled by three NF-kappaB binding sites was repressed by dexamethasone. The presence of only one GRE in the vicinity of the KB sites abolished this repression and allowed synergistic transcriptional activation by NF-kappaB and the glucocorticoid receptor (GR). The synergism identified here was not affected by the number of GREs, but strictly depends on the spacing between GREs and KB sites. These experiments reveal that the functional interplay between NF-kappaB and the GR also involves--dependent on the promoter context--synergistic stimulation of transcription. PMID- 12553733 TI - Expression of hepatocyte growth factor activator inhibitor type 2 (HAI-2) in human testis: identification of a distinct transcription start site for the HAI-2 gene in testis. AB - Hepatocyte growth factor activator inhibitor type 1 (HAI-1) and type 2 (HAI-2) are recently identified integral membrane Kunitz-type proteinase inhibitors. They have important regulatory roles in pericellular activation of hepatocyte growth factor/scatter factor (HGF/SF) which is critically involved in the development and regeneration of various tissues. Recent reports suggest that HGF/SF is also involved in testicular development and spermatogenesis. In this study, we analyzed the expression of HAIs in the testis. In human testis, HAI-2 was strongly expressed whereas HAI-1 mRNA was hardly detectable. Of interest was the observation that the mRNA size of HAI-2 was shorter in the testis (1.2 kb) than those in the other tissues such as placenta (1.5 kb). Subsequent experiments revealed that there are two major transcription start sites of the HAI-2 gene, which are -30 bp and -360 bp upstream from the translation initiation ATG codon. Although the latter site appeared to be mainly used in the placenta and other non testicular organs, only the former site is used in testis, resulting in the -300 bp shorter mRNA. An immunohistochemical study using a specific monoclonal antibody raised against human HAI-2 protein indicated that HAI-2 is expressed exclusively in primary spermatocytes. These results suggest a distinct regulation of HAI-2 gene expression in testis and that HAI-2 may play a role in the process of spermatogenesis. PMID- 12553734 TI - Identification and characterization of an IgG binding protein in the secretion of the rat coagulating gland. AB - A merocrine released protein (named 115k protein) was highly enriched from the secretion of the rat coagulating gland. The protein has a molecular mass of 115 kDa as calculated by SDS-PAGE under reducing conditions. Furthermore, the 115 kDa protein is glycosylated, and carries Man, GlcNAc, Gal, Fuc and sialic acid residues. For identification, N-terminal amino acid and nucleotide sequence analyses were performed. The sequences obtained showed 86 to 100% identity with human and mouse IgGFc binding proteins. The functional capacity of IgG binding of the 115 kDa protein was shown by overlay experiments, indicating its membership in the IgG binding protein family. PMID- 12553735 TI - Characterization of different reactive lysines in bovine heart mitochondrial porin. AB - Incubation of mitochondrial outer membrane porin with citraconic anhydride prior to treatment with fluorescein isothiocyanate (FITC) resulted in the labeling of a set of lysines located at a boundary between the water phase and lipid phase. The elution pattern of porin from the cation exchanger has been considered as indicative for the location of lysines. Electrical measurements after reconstitution of the modified protein in lipid bilayer membranes revealed that certain specific lysine residues are more susceptible to alterations. The innermost positive residues were only slightly influenced, while the outermost lysines exhibited a substantial change in channel properties. These results suggest the presence of critical charged residues in mitochondrial outer membrane porin that may be responsible for both the channel's selectivity and its voltage dependence. PMID- 12553736 TI - Determination of the mRNA sequence of cathepsin Y, a cysteine endopeptidase from rat spleen, 1 and confirmation of its ubiquitous expression. AB - A cysteine endopeptidase from rat spleen was purified, characterized and its gene cloned. This enzyme was originally recognized by its action of producing kinin potentiating peptide from a plasma protein. We named it cathepsin Y due to its localization, acidic pH optimum and the presence of the same set of active site amino acids as in other thiol cathepsins. Here we show the total sequence of the mRNA obtained by means of TaKaRa 5' Full RACE Core Set and complete the previously reported sequence. This completion of the mRNA sequence resulted in the omission of the strangely attached C-terminal peptide from cathepsin Y. PMID- 12553737 TI - Vascular calcification: good, bad or ugly? PMID- 12553738 TI - The contribution of nitric oxide to exercise hyperemia in the human forearm. AB - The contribution of nitric oxide (NO) to exercise-induced hyperemia is debated. Previous conclusions that nitric oxide synthase (NOS) inhibition reduces endothelium-dependent vasodilation during exercise hyperemia may be confounded by inhibitor-mediated increases in resting vascular tone. In this study, nine healthy participants performed wrist flexion exercise before and during intra arterial administration of the NOS-inhibitor NG-monomethyl-L-arginine (L-NMMA, 2 mg x min(-1)). Nine additional subjects performed this procedure while nitroprusside (0.2 microg x min(-1)) was co-infused with L-NMMA to maintain basal flow. Forearm blood flow was assessed with venous occlusion strain-gauge plethysmography at baseline, immediately after cessation of exercise, and continuously for 5 minutes thereafter. L-NMMA alone reduced resting flow by 26%, peak flow immediately after exercise by 20%, and integrated post-exercise hyperemic volume by 50% (all p < 0.05). Stabilization of resting vasodilator tone by nitroprusside eliminated the effects of L-NMMA on peak flow after exercise, yet L-NMMA still attenuated total hyperemic volume. In a time-control study of 12 subjects, there was no change in peak blood flow or hyperemic volume. This study indicates that NO is not a major regulator of peak limb blood flow measured immediately after cessation of dynamic exercise. The contribution of NO to exercise hyperemia is limited to the recovery period after exercise. PMID- 12553739 TI - A novel ultrasound method for evaluation of collateral development in limb ischemia. AB - Although clinical studies are underway to evaluate the effectiveness of angiogenesis, there are no validated, non-invasive methods to assess peripheral collateral development. This study was performed to validate a novel ultrasound based method of assessing collateral formation in a pig model of hindlimb ischemia. Ultrasonography of predefined ultrasound planes was performed on 12 pigs immediately after ligation of the right common femoral artery, and 7, 14, 28 and 42 days thereafter. A custom software program was used to evaluate both color Doppler (CD) and power Doppler (PD) images to generate flow indices. Collateral development was observed with ultrasound as early as 7 days post-arteriectomy and increased dramatically by 28 days. Areas of persistent ischemia resulting from inadequate collateral formation were easily quantified in all images. Collaterals detected on ultrasound were confirmed by angiography and histology, and tissue perfusion by a fluorescent microsphere method. As demonstrated with color and power Doppler measurements, collateral formation is initiated early after ischemic injury in this large juvenile animal model of angiogenesis. This non invasive method is useful to quantify blood flow, visualize angiogenesis and determine areas of persistent lower limb ischemia, and may have an important role in evaluating new approaches to modulate angiogenesis. PMID- 12553740 TI - Isolated internal jugular vein thrombosis: risk factors and natural history. AB - Deep venous thrombosis (DVT) involving the upper extremities, chest, and neck has become appreciated as a significant cause of morbidity, especially in individuals with central venous access devices (CVAD). Like DVT involving the leg and pelvic veins, axillo-subclavian vein thrombosis can result in pulmonary embolism, post thrombotic syndrome, and venous limb gangrene. Data relating to the natural history of internal jugular vein thrombosis (IJVT) is lacking. Risk factors, treatment patterns, and clinically relevant outcomes were retrospectively assessed in 74 consecutive patients with isolated IJVT. All patients (median age 66 years; range 36-80) had CVAD-associated IJVT, 22 (29.7%) had a history of cancer, and 14 (18.9%) had a history of prior DVT. Thirty-two (43.2%) were treated with systemic anticoagulation, 2 (2.7%) received a superior vena cava filter, and 40 (54.1%) received no specific therapy. Of the patients who underwent serial imaging, 7/40 (17.5%) had thrombus propagation. Two (2.7%) patients were diagnosed with pulmonary embolism, 8 (10.8%) were diagnosed with malignancy during a median follow-up of 20 months (range 18-24), and 3/53 (5.7%) suffered a recurrent DVT. Outcomes similar to those seen in leg DVT were observed. The most effective treatment strategy remains to be determined, but anticoagulant treatment was associated with a trend towards reduced all-cause mortality. PMID- 12553741 TI - Massive pulmonary embolism: a remarkable case and review of treatment. AB - Although the clinical syndrome following pulmonary embolism (PE) may be subtle, in the case of massive PE the severity of the clinical presentation typically makes the picture more obvious. If more than two lobar pulmonary arteries become obstructed, the hemodynamic and respiratory consequences are severe, and may be life threatening. We present an unusual case where a patient experienced transient hemodynamic collapse during valsalva, and a massive saddle pulmonary embolus was discovered incidentally. Current treatment options for massive PE are then discussed. PMID- 12553742 TI - Treating to goal: new strategies for initiating and optimizing lipid-lowering therapy in patients with atherosclerosis. AB - Abstract: The National Cholesterol Education Program (NCEP) guidelines prepared by the Adult Treatment Panel (ATP) provide clinicians with recommendations for the clinical management of abnormal blood cholesterol to reduce the risk of cardiovascular events. The recently updated NCEP guidelines have included a number of key amendments such as the recognition of low high-density lipoprotein cholesterol (HDL-C) as a risk factor for cardiovascular disease, while maintaining the focus of treatment on lowering low-density lipoprotein cholesterol (LDL-C) levels. Several agents can be used to modify the lipid profile in-line with the NCEP ATP-III recommendations, but hydroxymethyl glutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) remain the most effective and best-tolerated drugs for lowering LDL-C. The optimal time to initiate drug therapy had been in question, although recent studies suggest in-hospital initiation following admission with cardiovascular disease reduces the risk of recurrent events and improves long-term patient compliance. Inpatient physicians and nurses therefore play a pivotal role in influencing not only short-term management needs, but also the long-term recovery of their patients. This role should not be underestimated, as recent surveys have highlighted a significant problem of undertreatment in patients with documented atherosclerosis with regards to lipid-lowering and other cardioprotective therapy. Although reversing patient undertreatment will require clinicians to address a variety of issues, systematic in-hospital initiation of currently available lipid-lowering therapies in patients with atherosclerotic vascular disease is likely to have major benefits, reducing the occurrence of cardiovascular events and saving lives. Several statins, cholesterol absorption inhibitors, novel metabolic inhibitors, and combinations of agents are currently in clinical development and it is hoped that these will also assist clinicians in the important task of getting patients to recommended LDL-C and HDL-C levels. PMID- 12553743 TI - Carbon monoxide and bile pigments: surprising mediators of vascular function. AB - Heme oxygenase (HO) catalyzes the degradation of heme to CO, iron, and biliverdin. Biliverdin is subsequently metabolized to bilirubin by the enzyme biliverdin reductase. Although long considered irrelevant byproducts of heme catabolism, recent studies indicate that CO and the bile pigments biliverdin and bilirubin may play an important physiological role in the circulation. The release of CO by vascular cells may modulate blood flow and blood fluidity by inhibiting vasomotor tone, smooth muscle cell proliferation, and platelet aggregation. CO may also maintain the integrity of the vessel wall by directly blocking vascular cell apoptosis and by inhibiting the release of pro-apoptotic inflammatory cytokines from the vessel wall. These effects of CO are mediated via multiple pathways, including activation of soluble guanylate cyclase, potassium channels, p38 mitogen-activated protein kinase, or inhibition of cytochrome P450. In addition, the release of bile pigments may serve to sustain vascular homeostasis by protecting vascular cells from oxidative stress and by inhibiting the adhesion and infiltration of leukocytes into the vessel wall. Induction of HO 1 gene expression and the subsequent release of CO and bile pigments are observed in numerous vascular disorders and may provide an important adaptive mechanism to preserve homeostasis at sites of vascular injury. Thus, the HO-catalyzed formation of CO and bile pigments by vascular cells may function as a critical endogenous vasoprotective system. Moreover, pharmacological or genetic approaches targeting HO-1 to the vessel wall may represent a novel therapeutic approach in treating vascular disease. PMID- 12553744 TI - L-arginine in cardiovascular disease: dream or reality? AB - L-arginine is the substrate for endothelial nitric oxide synthase (eNOS), and the precursor for the synthesis of nitric oxide (NO). This amino acid exerts a number of actions in the cardiovascular system, mainly through the production of NO. However, it also has a number of NO-independent properties, such as the ability to regulate blood and intracellular pH and the effect on the depolarization of endothelial cell membranes. It also has antihypertensive and antioxidant properties, it influences blood viscosity and the coagulation/fibrinolysis system, and it affects the metabolism of glucose, lipids and proteins. L-arginine influences a number of atherosclerosis risk factors such as hypercholesterolemia, hypertension and smoking, improving endothelial function in these patients. However, it does not affect endothelial function in patients with diabetes mellitus. The role of L-arginine in coronary artery disease is still controversial, but it seems that oral or parenteral administration of this amino acid restores endothelial function in the brachial artery and improves coronary microcirculation. The role of L-arginine in heart failure is currently under investigation, and the first results are rather hopeful. In conclusion, L arginine seems to provide a hopeful prospect for the treatment of cardiovascular diseases. However, more data derived from large-scale prospective studies evaluating the effects of long-term treatment with L-arginine are needed. PMID- 12553745 TI - The pathophysiology of erectile dysfunction related to endothelial dysfunction and mediators of vascular function. AB - The incidence of erectile dysfunction increases with diabetes, hypertension, hypercholesterolaemia, cardiovascular disease and renal failure. All these conditions are associated with endothelial dysfunction. This review addresses the pathophysiology of erectile dysfunction with a special focus on new insights into nitric oxide (NO)-mediated pathways, oxidative stress and parallels to endothelial dysfunction. NO appears to be the key mediator promoting endothelium derived vasodilation and penile erection. The possibility is discussed that elevated plasma concentrations of asymmetrical dimethylarginine (ADMA), an endogenous NO synthase inhibitor, may provide an additional pathomechanism for various forms of erectile dysfunction associated with cardiovascular risk factors and disease. Likewise, the role of endothelium-derived factors mediating NO independent pathways is evaluated. PMID- 12553746 TI - Endothelial dysfunction and atherothrombosis in mild hyperhomocysteinemia. AB - Mildly elevated plasma homocysteine levels are an independent risk factor for atherothrombotic vascular disease in the coronary, cerebrovascular, and peripheral arterial circulation. Endothelial dysfunction as manifested by impaired endothelium-dependent regulation of vascular tone and blood flow, by increased recruitment and adhesion of circulating inflammatory cells to the endothelium, and by a loss of endothelial cell antithrombotic function contributes to the vascular disorders linked to hyperhomocysteinemia. Increased vascular oxidant stress through imbalanced thiol redox status and inhibition of important antioxidant enzymes by homocysteine results in decreased bioavailability of the endothelium-derived signaling molecule nitric oxide via oxidative inactivation. This plays a central role in the molecular mechanisms underlying the effects of homocysteine on endothelial function. Supplementation of folic acid and vitamin B12 has been demonstrated to be efficient in lowering mildly elevated plasma homocysteine levels and in reversing homocysteine-induced impairment of endothelium-dependent vasoreactivity. Results from ongoing intervention trials will determine whether homocysteine-lowering therapies contribute to the prevention and reduction of atherothrombotic vascular disease and may thereby provide support for the causal relationship between hyperhomocysteinemia and atherothrombosis. PMID- 12553747 TI - Toward a new blood vessel. AB - Strategies to treat atherosclerotic coronary artery disease include coronary artery bypass grafting (CABG), in which grafts are used to bypass atherosclerotic vessels and restore blood flow to the ischemic myocardium. The grafts used include healthy arteries or veins harvested from a separate site. Results with arterial grafts have been superior to venous grafts; promoting the practice of total arterial revascularization using only arterial grafts. Suitable arterial grafts, however, are scarce and harvest procedures add to morbidity and cost. Tissue engineering combines the principles of engineering with life sciences for the development of biological substitutes and restore, maintain or improve tissue function. Advances in this field have included the development of tissue engineered blood vessels, with the potential to serve as arterial grafts, conduits or fistulae. This review describes the history of tissue engineering arteries, the techniques used, and progress to date. The source of cells and the future direction of this field are explored. PMID- 12553748 TI - Images in vascular medicine. Traumatic aortic transection. PMID- 12553749 TI - Characterization of soluble polymer supported organic compounds by LC/electrospray ionization MS toward a complete automation of the liquid-phase process in combinatorial chemistry. AB - In the case of a soluble polymer supported synthesis, the tuning of the support solubility depending on the solvent offers new opportunities for combinatorial chemistry for two reasons. First, organic syntheses in solution are generally easily translated on a soluble polymer supported substrate. Second, complete automation of the three successive steps of combinatorial chemistry (synthesis, analytical control, biological test) is approached without releasing the synthesized compound from the support. We report in this paper the preliminary results toward automation of both soluble polymer supported syntheses and their subsequent monitoring by ESI mass spectrometry. PMID- 12553750 TI - Patterned monolayer/polymer films for analysis of dilute or salt-contaminated protein samples by MALDI-MS. AB - This paper describes a surface science/mass spectrometry effort to develop and characterize a patterned gold surface that serves as a MALDI sample platform capable of concentrating and purifying proteins. Using microcontact printing, small (200-microm diameter) hydrophilic spots of bare gold or chemically anchored poly(acrylic acid) (PAA) are patterned at 5-mm intervals in a hydrophobic field consisting of a self-assembled monolayer of hexadecanethiol. Building on recent innovations by others, the small hydrophilic spots concentrate the sample to achieve good reproducibility and high sensitivity in the MALDI signal. One of the key features in this work is the combination of the high density of carboxylate groups in PAA with a small spot size to afford both concentration and purification of proteins via ionic interactions. This translates into detection limits for salt-contaminated proteins that are 20-100 times lower (low femtomole) than those reported for previous polymer- or monolayer-modified MALDI probes (using proteins in the 3-15-kDa range). Reflectance FT-IR spectroscopy and ellipsometry were used to determine the amount of protein adsorbed to a PAA modified sample plate as a function of pH and salt concentration. Amide absorbances in IR spectra correlate well with MALDI-MS signals measured after addition of 2,5-dihydroxybenzoic acid as a matrix. PMID- 12553751 TI - Uniformly sized molecularly imprinted polymer for (S)-nilvadipine. Comparison of chiral recognition ability with HPLC chiral stationary phases based on a protein. AB - Uniformly sized molecularly imprinted polymers (MIPs) for (S)-nilvadipine have been prepared by a multistep swelling and polymerization method using methacrylic acid, 2-(trifluoromethyl)acrylic acid, 2-vinylpyridine, or 4-vinylpyridine (4 VPY) as a functional monomer and ethylene glycol dimethacrylate (EDMA) as a cross linker. The chiral recognition abilities of the MIPs for nilvadipine and other dihydropyridine calcium antagonists were evaluated using a mixture of sodium phosphate buffer (or water) and acetonitrile or only acetonitrile as the mobile phase. The (S)-nilvadipine-imprinted 4-VPY-co-EDMA polymers gave the highest resolution for nilvadipine among the MIPs prepared. In addition, the enantioseparation of nilvadipine was attained using the (S)-nilvadipine-imprinted EDMA polymers, without use of a functional monomer. 1H NMR and molecular modeling studies suggested a one-to-one hydrogen-bonding-based complex formation of (S) nilvadipine with 4-VPY in chloroform. These results reveal that the (S) nilvadipine-imprinted EDMA polymers could recognize the template molecule by its molecular shape, and that in addition to this recognition, hydrophobic and hydrogen-bonding interactions seems to play important roles in the retention and chiral recognition of nilvadipine on the 4-VPY-co-EDMA polymers in hydroorganic mobile phases. By optimizing chromatographic conditions such as column temperature and flow rate, the baseline separation of nilvadipine enantiomers was attained with a short analysis time and with a column efficiency comparable to commercially available chiral stationary phases based on a protein, such as ovomucoid or alpha1-acid glycoprotein. PMID- 12553752 TI - HPLC of monolayer-protected gold nanoclusters. AB - Polydisperse samples of Au nanoparticles protected with monolayers of hexanethiolate ligands (C6 MPCs) and with mixed monolayers of hexanethiolate and mercaptoundecanoic acid (C6/MUA MPCs) have been chromatographically separated using C8 120-A columns and acetone/ toluene mobile phase. The spectral details of eluted peaks and of quantized double-layer charging features in the differential pulse voltammetry of collected fractions were used to show that the elution orders of C6 MPC mixtures and of C6/MUA MPC mixtures were different. For C6 MPCs, the smallest MPCs were eluted first, whereas the smallest C6/MUA MPCs were eluted last. The reversal of order of elution was rationalized in terms of intermolecular interactions with the stationary phase, dominant for the C6 MPC, being suppressed by the heightened polarity of the monolayer surface of the C6/MUA MPCs, making a size exclusion mechanism dominant. The range of apparent core diameters of the separated nanoparticles was 1.3-2 nm. PMID- 12553753 TI - Effect of dextran as a run buffer additive in drug-protein binding studies using capillary electrophoresis frontal analysis. AB - The study of drug-protein interactions by capillary electrophoresis frontal analysis requires establishment of a sufficient mobility difference between the mobility of the ligand and protein. The potential utility of dextran as a run buffer additive to manipulate the electrophoretic mobilities of low molecular weight ligands and protein in capillary electrophoresis frontal analysis binding studies was assessed. It was demonstrated that dextran was effective in improving the separation between the ligands warfarin and flurbiprofen and human serum albumin. Separation of ligand and protein increased with the concentration of added dextran (0-7.5% (w/w)), while molecular weight of the additive (70,000 2,000,000) only had a minor effect. The effect of dextran addition on viscosity and electrophoretic and electroosmotic mobilites was systematically studied. Optimal frontal analysis settings were a compromise between achieving satisfactory separation and acceptable analysis times without loss of plateau peak conditions. No effect of dextran upon the drug-human serum albumin interactions could be detected for the model ligands. Introduction of dextran into the electrophoresis buffer expands the applicability of capillary electrophoresis frontal analysis in drug research to binding interactions between proteins and low molecular weight ligands possessing similar electrophoretic mobilities. PMID- 12553754 TI - On-line hyphenation of capillary isoelectric focusing and capillary gel electrophoresis by a dialysis interface. AB - An on-line two-dimensional (2D) capillary electrophoresis (CE) system consisting of capillary isoelectric focusing (CIEF) and capillary gel electrophoresis (CGE) was introduced. To validate this 2D system, a dialysis interface was developed by mounting a hollow fiber on a methacrylate resin plate to hyphenate the two CE modes. The two dimensions of capillary shared a cathode fixated into a reservoir in the methacrylate plate; thus, with three electrodes and only one high-voltage source, a 2D CE framework was successfully established. A practical 2D CIEF-CGE experiment was carried out to deal with a target protein, hemoglobin (Hb). After the Hb variants with different isoelectric points (pIs) were focused in various bands in the first-dimension capillary, they were chemically mobilized one after another and fed to the second-dimension capillary for further separation in polyacrylamide gel. During this procedure, a single CIEF band was separated into several peaks due to different molecular weights. The resulting electrophoregram is quite different from that of either CIEF or CGE; therefore, more information about the studied Hb sample can be obtained. PMID- 12553755 TI - Quantification of cell and cellulase mass concentrations during anaerobic cellulose fermentation: development of an enzyme-linked immunosorbent assay-based method with application to Clostridium thermocellum batch cultures. AB - A methodology was developed to determine the mass concentrations of cellulase and cells applicable to studies of microbial cellulose utilization in systems for which a substantial fraction of cellulase is cell-associated. Antibodies raised against a 14-amino acid synthetic peptide with sequence taken from the cohesin domain of the scaffoldin protein of Clostridium thermocellum ATCC 27405 were used to develop an indirect ELISA protocol. Six cellulase calibration standards were prepared using affinity digestion (Morag, E.; Bayer, E. A.; Lamed, R. Enzyme Microb. Technol. 1992, 14, 289-292.). These included supernatant and pellet samples from an Avicelgrown culture with fractional cellulose conversion (X) = 0.98, as well as supernatant, pellet, cell-associated, and cellulose-associated samples from an Avicel-grown culture with X = 0.8. All six standards displayed a very similar absorbance versus concentration relationship when subjected to ELISA, essentially identical SDS-PAGE banding patterns, and similar cellulase specific activity in relation to both other purified cellulase preparations and crude samples. Coefficients of variation for cellulase concentration measurements were 5.2% for supernatant samples and 5.9% for pellet samples. The ELISA method was applied to batch cultures of C. thermocellum grown on Avicel. Cell concentration was calculated from the pellet protein concentration and the cell protein fraction of a cellobiose-grown control. Two alternative methods appeared to overpredict the cell concentration and were not capable of quantifying cells as distinct from cellulase. Cellulase protein production by Avicel-grown batch cultures represented approximately 20% of cell mass exclusive of cellulase. It is concluded that the reported protocols establish a reasonable methodological basis for quantitative determination of the mass concentration of cellulase protein produced by C. thermocellum and for calculation of cell mass concentration as distinct from cellulase concentration. PMID- 12553756 TI - Chemically assisted laser ablation ICP mass spectrometry. AB - A new laser ablation technique combined with a chemical evaporation reaction has been developed for elemental ratio analysis of solid samples using an inductively coupled plasma mass spectrometer (ICPMS). Using a chemically assisted laser ablation (CIA) technique developed in this study, analytical repeatability of the elemental ratio measurement was successively improved. To evaluate the reliability of the CLA-ICPMS technique, Pb/U isotopic ratios were determined for zircon samples that have previously been analyzed by other techniques. Conventional laser ablation for Pb/U shows a serious elemental fractionation during ablation mainly due to the large difference in elemental volatility between Pb and U. In the case of Pb/U ratio measurement, a Freon R-134a gas (1,1,1,2-tetrafluoroethane) was introduced into the laser cell as a fluorination reactant. The Freon gas introduced into the laser cell reacts with the ablated sample U, and refractory U compounds are converted to a volatile U fluoride compound (UF6) under the high-temperature condition at the ablation site. This avoids the redeposition of U around the ablation pits. Although not all the U is reacted with Freon, formation of volatile UF compounds improves the transmission efficiency of U. Typical precision of the 206Pb/238U ratio measurement is 3-5% (2sigma) for NIST SRM 610 and Nancy 91500 zircon standard, and the U-Pb age data obtained here show good agreement within analytical uncertainties with the previously reported values. Since the observed Pb/U ratio for solid samples is relatively insensitive to laser power and ablation time, optimization of ablation conditions or acquisition parameters no longer needs to be performed on a sample to-sample basis. PMID- 12553757 TI - Determination of polycyclic aromatic hydrocarbons with molecular weight 300 and 302 in environmental-matrix standard reference materials by gas chromatography/mass spectrometry. AB - An analytical approach based on gas chromatography/ mass spectrometry (GC/MS) is presented for the measurement of polycyclic aromatic hydrocarbons with molecular weight (MW) 300 and 302 in environmental samples. Three different GC stationary phases [5% and 50% phenyl methylpolysiloxane and dimethyl (50% liquid crystalline) polysiloxane] were compared, and retention indexes (RI) are given for 23 individual MW 302 isomers. Identification of MW 300 and 302 isomers in four environmental-matrix Standard Reference Materials (SRMs) (SRM 1597, coal tar extract; SRM 1648 and SRM 1649a, air particulate matter; and SRM 1941, marine sediment) was based on the comparison of RI data and mass spectra from authentic standards. Dibenzo[a,l]pyrene, which is of considerable interest because of its high carcinogenicity, was identified and quantified in the four environmental matrix SRMs. A total of 23 isomers of MW 302 and four isomers of MW 300 were quantified in four different environmental-matrix SRMs, and the results are compared to previously reported results based on liquid chromatography with fluorescence detection. PMID- 12553758 TI - Enhancement of immunocomplex detection and application to assays for DNA adduct of benzo[a]pyrene. AB - The stability of antibody and formation of immunocomplexes are essential to high sensitivity capillary electrophoresis immunoassays (CEIA). However, little attention has been paid to enhancing or maintaining immunocomplex formation and antibody stability to improve the performance of CEIA. We report here the use of nonspecific proteins, such as bovine serum albumin (BSA) and rabbit immunoglobulin (rIgG), to enhance immunocomplex formation and to stabilize antibodies and immunocomplexes for immunoassays. Complexes between DNA adducts of benzo[a]pyrenediol epoxide (BPDE) and their antibodies were examined using capillary electrophoresis with laser-induced fluorescence detection (CE-HF). A tetramethylrhodamine (TMR)-labeled single-stranded oligonucleotide (16-mer) containing a single BPDE adduct was used as a fluorescent probe to study its immunocomplexes with a monoclonal antibody (8E11). To examine the formation of larger complexes, a TMR-labeled secondary antibody (anti-mouse), a primary antibody (mouse monoclonal antibody 5D11), and BPDE adducts in cellular DNA were used. We demonstrate that the use of nonspecific proteins stabilized the antibody and greatly enhanced the formation and stability of the immunocomplexes, resulting in substantial improvements in the detection limit (10-fold) and the reproducibility of the analysis. Another advantageous consequence of the stabilization was a 150-fold reduction of the concentration of the antibody needed for the immunoassay, resulting in reduced background and cost. We successfully applied this technique to the determination of DNA adducts of BPDE using a competitive immunoassay. The results from both small complexes (between a primary antibody and an oligonucleotide) and larger complexes (among a secondary antibody, a primary antibody, and cellular DNA) indicate that the technique can be extended to other immunoassays. We suggest that nonspecific proteins may assist the formation and stabilization of antibody-antigen complexes by maintaining the correct conformation of the antibody and antigen for optimum binding. PMID- 12553759 TI - Fluorophoric assay for the high-throughput determination of amidase activity. AB - An assay has been developed for the high-throughput identification of amidase activity. Amines released from the enzyme-catalyzed hydrolysis of corresponding amides were detected by the formation of a fluorescent dye by coupling with 4 nitro-7-chloro-benzo-2-oxa-1,3-diazole (NBD-CI). Using this format, 22 lipases and esterases were tested for their ability to hydrolyze aromatic substituted N acylamines in a microtiter plate format. Identified active enzymes were further characterized toward a broad range of compounds to determine the influence of substrate structure on activity. For recombinantly produced esterases, it could be shown that the assay works with high reproducibility and sensitivity, even in the presence of amino acids and proteins present in culture media and cell debris. PMID- 12553760 TI - Quantification of o,o'-dityrosine, o-nitrotyrosine, and o-tyrosine in cat urine samples by LC/ electrospray ionization-MS/MS using isotope dilution. AB - Quantification of o-tyrosine, o-nitrotyrosine, and o,o'dityrosine from cat urine samples was achieved by LC/ electrospray ionization-MS/MS (LC/ESI-MS/MS) using an isotope dilution technique in multiple reaction monitoring mode before butylation of o,o'-dityrosine and after butylation of o-tyrosine and o-nitrotyrosine. This novel approach of amino acids butylation enhanced the MS response by a factor of 7-fold for o-tyrosine and 6-fold for o-nitrotyrosine and decreased the overall chemical background noise. Butylation of o,o'-dityrosine resulted in a lower MS response as a result of the formation of both mono- and doubly butylated species. The mean recovery for the oxidized amino acids was estimated at 73 +/- 2%. The limits of quantitation of NO2-Tyr butyl ester, o-Tyr butyl ester, and di-Tyr in cat urine samples were calculated at 14.5, 28.2 and 140.9 nM, respectively. The oxidized amino acids levels in cat urine extracts ranged from 157 to 250 ng/day for o-Tyr and from 3,289 to 11,803 ng/day for di-Tyr. NO2-Tyr was found in only two urine extracts at levels below 58 ng/day. A certain trend of correlation was observed between o,o'-dityrosine and o-tyrosine when comparing these values against their respective creatinine amounts. A comparison of the data gathered from the ThermoFinnigan TSQ 7000 and Micromass Q-TOF instruments revealed several advantages of using the Q-TOF regarding the exact mass measurement, a lower ion suppression effect and the possibility to perform analyses in full scan product ion mode. These results demonstrate that a Q-TOF instrument can be a good alternative to classical triple quadrupole for quantitative purposes on a relatively small linear dynamic range (4 orders of magnitude for the Q-TOF, as compared to 6 for the triple quadrupole). PMID- 12553761 TI - Solid-phase extraction NMR studies of chromatographic fractions of saponins from Quillaja saponaria. AB - The saponin mixture QH-B from the tree Quillaja saponaria var. Molina was fractionated by RP-HPLC in several steps. The fractions were analyzed by solid phase extraction NMR (SPE-NMR), a technique combining the workup by solid-phase extraction with on-line coupling to an NMR flow probe. Together with MALDI-TOF mass spectrometry and comparison with chemical shifts of similar saponins, the structures of both major and minor components in QH-B could be obtained. The procedure described is a simple method to determine the structure of components in a complex mixture. The two major fractions of the mixture were found to contain at least 28 saponins, differing in the carbohydrate substructures. Eight of these have not previously been determined. The 28 saponins formed 14 equilibrium pairs by the migration of an O-acyl group between two adjacent positions on a fucosyl residue. PMID- 12553762 TI - A miniature biochip system for detection of aerosolized Bacillus globigii spores. AB - The feasibility of using a novel detection scheme for the analysis of biological warfare agents is demonstrated using Bacillus globigii spores, a surrogate species for Bacillus anthracis. In this paper, a sensitive and selective enzyme linked immunosorbent assay using a novel fluorogenic alkaline phosphatase substrate (dimethylacridinone phosphate) is combined with a compact biochip detection system, which includes a miniature diode laser for excitation. Detection of aerosolized spores was achieved by coupling the miniature system to a portable bioaerosol sampler, and the performance of the antibody-based recognition and enzyme amplification method was evaluated. The bioassay performance was found to be compatible with the air sampling device, and the enzymatic amplification was found to be an attractive amplification method for detection of low spore concentrations. The combined portable bioaerosol sampler and miniature biochip system detected 100 B. globigii spores, corresponding to 17 aerosolized spores/L of air. Moreover, the incorporation of the miniature diode laser with the self-contained biochip design allows for a compact system that is readily adaptable to field use. In addition, these studies have included investigations into the tradeoff between assay time and sensitivity. PMID- 12553763 TI - Surfactant-induced electrophoretic mobility of intact cellulose polymer derivatives. AB - Chemically modified celluloses are mobilized in electric fields as complexes with charged surfactants. Depending on the nature of a participating amphiphile and the type of substitution for a given class of cellulose derivative, different kinds of interactions take place. The various migration mechanisms are described through determination of adsorption isotherms followed by nonlinear model fitting of simple semiempirical migration models. Various anionic surfactants differing mainly in the hydrocarbon skeleton were employed as mobility inducers along with an uncharged and carbohydrate-based detergent. A clear distinction between differently substituted celluloses regarding mobility and selectivity is observed, and capillary electrophoresis shows to be a technique capable of separating mixtures of intact cellulose derivatives independent of molecular weight or hydrodynamic radius. PMID- 12553764 TI - Microfabricated device for DNA and RNA amplification by continuous-flow polymerase chain reaction and reverse transcription-polymerase chain reaction with cycle number selection. AB - We have developed a high-throughput microfabricated, reusable glass chip for the functional integration of reverse transcription (RT) and polymerase chain reaction (PCR) in a continuous-flow mode. The chip allows for selection of the number of amplification cycles. A single microchannel network was etched that defines four distinct zones, one for RT and three for PCR (denaturation, annealing, extension). The zone temperatures were controlled by placing the chip over four heating blocks. Samples and reagents for RT and PCR were pumped continuously through appropriate access holes. Outlet channels were etched after cycles 20, 25, 30, 35, and 40 for product collection. The surface-to-volume ratio for the PCR channel is 57 mm(-1) and the channel depth is 55 microm, both of which allow very rapid heat transfer. As a result, we were able to collect PCR product after 30 amplification cycles in only 6 min. Products were collected in 0.2-mL tubes and analyzed by agarose gel electrophoresis and ethidium bromide staining. We studied DNA and RNA amplification as a function of cycle number. The effect of the number of the initial DNA and RNA input molecules was studied in the range of 2.5 x 10(6) - 1.6 x 10(8) and 6.2 x 10(6) - 2 x 10(8), respectively. Successful amplification of a single-copy gene (beta-globin) from human genomic DNA was carried out. Furthermore, PCR was performed on three samples of DNA of different lengths (each of 2-microL reaction volume) flowing simultaneously in the chip, and the products were collected after various numbers of cycles. Reverse transcription was also carried out on four RNA samples (0.7-microL reaction volume) flowing simultaneously in the chip, followed by PCR amplification. Finally, we have demonstrated the concept of manually pumped injection and transport of the reaction mixture in continuous-flow PCR for the rapid generation of amplification products with minimal instrumentation. To our knowledge, this is the first report of a monolithic microdevice that integrates continuous-flow RT and PCR with cycle number selection. PMID- 12553765 TI - Modified carbon surfaces as "organic electrodes" that exhibit conductance switching. AB - Glassy carbon (GC) surfaces modified with monolayers of biphenyl and nitrobiphenyl molecules were examined as voltammetric electrodes for ferrocene, benzoquinone, and tetracyanoquinodimethane electrochemistry in acetonitrile. The modified electrodes exhibited slower electron transfer than unmodified GC, by factors that varied with the monolayer and redox system. However, after a negative potential excursion to approximately -2.0 V versus Ag+/Ag, the modified electrodes exhibited much faster electron-transfer kinetics, approaching those observed on unmodified GC. The effect is attributed to an apparently irreversible structural change in the biphenyl or nitrobiphenyl monolayer, which increases the rate of electron tunneling. The transition to the "ON" state is associated with electron injection into the monolayer similar to that observed in previous spectroscopic investigations and causes a significant decrease in the calculated HOMO-LUMO gap for the monolayer molecule. Once the monolayer is switched ON, it supports rapid electron exchange with outer-sphere redox systems, but not with dopamine, which requires adsorption to the GC surface. The increase in electron transfer rate with electron injection is consistent with an increase in electron tunneling rate through the monolayer, caused by a significant decrease in tunneling barrier height. The ON electrode can reduce biphenyl- or nitrobiphenyldiazonium reagent in solution to permit formation of a second modification layer of biphenyl or nitrobiphenyl molecules. This "double derivatization" procedure was used to prepare tetraphenyl- and nitrotetraphenyl modified electrodes, which exhibit significantly slower electron transfer than their biphenyl and nitrobiphenyl counterparts. A "switching" electrode may have useful properties for electroanalytical applications and possibly in electrocatalysis. In addition, the ON state represents an "organic electrode" in which electron transfer occurs at an interface between an organic conductor and a solution rather than an interface between a solution and a metal or carbon electrode. PMID- 12553766 TI - On-chip contactless four-electrode conductivity detection for capillary electrophoresis devices. AB - In this contribution, a capillary electrophoresis microdevice with an integrated on-chip contactless four-electrode conductivity detector is presented. A 6-cm long, 70-microm-wide, and 20-microm-deep channel was etched in a glass substrate that was bonded to a second glass substrate in order to form a sealed channel. Four contactless electrodes (metal electrodes covered by 30-nm silicon carbide) were deposited and patterned on the second glass substrate for on-chip conductivity detection. Contactless conductivity detection was performed in either a two- or a four-electrode configuration. Experimental results confirmed the improved characteristics of the four-electrode configuration over the classical two-electrode detection setup. The four-electrode configuration allows for sensitive detection for varying carrier-electrolyte background conductivity without the need for adjustment of the measurement frequency. Reproducible electrophoretic separations of three inorganic cations (K+, Na+, Li+) and six organic acids are presented. Detection as low as 5 microM for potassium was demonstrated. PMID- 12553767 TI - A two-channel microfluidic sensor that uses anodic electrogenerated chemiluminescence as a photonic reporter of cathodic redox reactions. AB - This paper describes a new approach for sensing electrochemically active substrates in microfluidic systems. This two-electrode sensor relies on electrochemical detection at one electrode and electrogenerated chemiluminescent (ECL) reporting at the other. Each microfabricated indium tin oxide electrode is located in a separate microfluidic channel, but the channels are connected downstream of the electrodes to maintain a complete electrical circuit. Because of laminar flow, there is no bulk mixing of the fluids in the detecting and reporting channels. This approach allows the ECL reaction to be physically and chemically decoupled from the sensing channel of the device, which greatly expands the number of analytes that can be detected. However, because the cathode and anode are connected, electron-transfer processes occurring at the sensing electrode are electrically coupled to the ECL reaction. Charge balance permits the ECL light output to be quantitatively correlated to electrochemical reductions at the cathode. The system is used to detect Fe(CN)6(3-), Ru(NH3)6(3+), and benzyl viologen and report their presence via Ru(bpy)3(2+) (bpy = bipyridine) luminescence. Each different redox target initiates ECL at a unique potential bias related to its standard redox potential. The influence of the concentrations of Ru(bpy)3(2+) and the target analytes is discussed. PMID- 12553768 TI - Application of disorganized monolayer films on gold electrodes to the prevention of surfactant inhibition of the voltammetric detection of trace metals via anodic stripping of underpotential deposits: detection of copper. AB - Development of an approach to prevention of electrode surface fouling by surfactants in samples is demonstrated. Spontaneously adsorbed monolayer systems employing short alkyl chains and bulky end groups are used to form porous disorganized monolayers on gold electrodes. Detection of copper by stripping of underpotential deposits formed at electrodes modified with disorganized films of mercaptoethanesulfonate (MES), mercaptopropanesulfonate, mercaptoacetic acid, and mercaptopropanoic acid was possible, and to a much lesser extent at aminoethanethiol and L-cysteine films. Use of short deposition times in conjunction with linear sweep anodic stripping voltammetry allowed detection of Cu2+ ions down to 1 x 10(-6) M in sulfuric acid solution, using underpotential deposition as the deposition step of the procedure. Calibration graphs were linear in the concentration range (1-80) x 10(-6) M Cu2+ using 15-s deposition at 0.00 V versus Ag/AgCl. The surfactants Tween 20, Tween 80, and Triton X-100 were found to have no affect on detection of Cu2+ ions in the calibration curve concentration range using MES-modified gold electrodes, whereas at unmodified gold electrodes very severe attenuation of the detection capability was manifested. The average slope for all calibration curves at the MES-modified electrode in the absence and presence of the surfactants at two different concentration levels was 0.0710 +/- 0.0024 microA microM(-1); in contrast, the slope of the calibration line at uncoated gold electrodes in the presence of surfactant was 0.0268 microA microM(-1). These results indicate the excellent ability of a disorganized, porous monolayer for prevention of fouling of the electrode surface by the surfactants. PMID- 12553769 TI - Electrochemiluminescence of luminol in alkaline solution at a paraffin impregnated graphite electrode. AB - The behavior of luminol electrochemiluminescence (ECL) at a paraffin-impregnated graphite electrode (PIGE) at different applied potentials was studied. Five ECL peaks were observed at 0.31, 0.59, 1.09, 1.54, and -0.58 V versus SCE, respectively, being related to potential scan direction and ranges, N2, O2, pH of the solution, and KCl concentration. The emission spectra of various ECL peaks at different potentials showed that all ECL peaks were initiated by luminol reactions. X-ray diffraction demonstrated that a simple mixture was formed between graphite and paraffin. The fluorescence spectra on the surface of the PIGE suggested that certain groups on the graphite were oxidized when the positive potential was applied to the electrode. In the presence of O2, three main ECL peaks were obtained in 0.1 mol/L KCl at pH 12.2. The ECL peak at 0.59 V with a shoulder is likely due to the reaction of luminol radicals with O2 and further electrooxidation of luminol radicals. The ECL peak at 1.54 V was suggested to be due to the electrooxidation of OH- to HO2- at higher potential and then to O2-, which reacted with luminol to produce light emission. Moreover, the oxygen-containing functional groups formed by the oxidation of the surface of the graphite electrode might enhance the ECL. At -0.58 V, the dissolved oxygen in solution was reduced to HO2-, resulting in light emission. At a potential higher than 1.64 V, ClO- was formed, leading to a broad emission wave and enhancement of the ECL peak at -0.58 V upon the reversal scan. Under nitrogen atmosphere, an ECL peak appeared at 1.09 V. At this potential, OH- was oxidized to O2, followed by the reaction with luminol to generate light emission. At pH 13.2 or 0.5 mol/L KCl, the shoulder of the ECL peak at 0.59 V became an ECL peak at 0.31 V. The conversion of luminol radicals into excited 3-aminophthalate may undergo two routes. Under these conditions, two routes might proceed at a different rate to form another ECL peak. It is concluded that luminol ECL could be readily excited by various oxygen-containing species electrogenerated at different applied potentials. Three strong ECL peaks obtained at different potentials on the PIGE might be of a potential to improve analytical selectivity and sensitivity for the detection of some analytes. PMID- 12553770 TI - Optical sensor for amine vapors based on dimer-monomer equilibrium of indium(III) octaethylporphyrin in a polymeric film. AB - A novel transduction chemistry for the development of a polymer film-based optical sensor that responds reversibly to gas-phase amine species at sub-ppm levels is described. The sensor is based on the equilibrium of a indium(III) octaethylporphyrin hydroxide ion-bridged dimer species with corresponding monomeric porphyrins within a thin poly(vinyl chloride) film as a function of the level of volatile amine in the surrounding gas phase. The presence of amines causes the dimeric species to be converted to monomer via the ligation of the amine with the In(III) center of the porphyrin structure. This yields a significant change in the visible absorption spectrum of the film, with a decrease in the intensity of the Soret band corresponding to the dimer (lambdamax = 390 nm) and a concomitant increase in the Soret band for the monomer lambdamax = 406-408 nm). Response to different amines is based on their relative partition coefficient into the polymer film and their strength of axial ligation reactions, with a selectivity pattern of 1-butylamine > 1-propylamine > pyridine > triethylamine > ethylamine > methylamine > diethylamine > tert-butylamine > ammonia. It is further shown that a significant concentration of dimeric species within the polymer film can only be achieved if appropriate amounts of lipophilic anionic sites are also incorporated into the polymer in the form of a tetraphenylborate derivative and the resulting film is equilibrated briefly with water prior to gas-phase measurements. With optimized film compositions, 1 butylamine can be detected in the gas phase to levels approaching 0.1 ppm, while less lipophilic ammonia can be monitored down to 10 ppm, with fully reversible responses to each species. A simple mathematical model for the response of the amine sensor is presented and shown to predict the optical behavior observed. PMID- 12553771 TI - Nonaqueous electrophoresis microchip separations: conductivity detection in UV absorbing solvents. AB - The use of organic solvents in microfabricated capillary electrophoresis (CE) devices is demonstrated in connection with the separation of aliphatic amines in pure dimethylformamide, dimethylacetamide, dimethyl sulfoxide, or propylene carbonate media. Contactless conductivity detection is employed for monitoring the separated solutes in these UV-absorbing solvents. The effect of the physicochemical properties of the organic solvent upon the migration behavior is investigated. The apparent mobility increases nearly linearly with the reciprocal of the solvent viscosity, while the electroosmotic mobility increases in a linear fashion with the dielectric constant/ viscosity ratio. Some deviation from theoretical predictions is observed using propylene carbonate. The nonaqueous CE microchip offers high separation efficiency, reflected in plate numbers ranging from 93,680 to 127,680, using a separation voltage of +3,000 V, a dimethylformamide medium, and a contactless conductivity detection. Experimental parameters affecting the analytical performance of the nonaqueous CE/conductivity microchip are examined. Calibration and precision experiments indicate a linear and reproducible response. Such use of organic solvents can benefit microchip separations through extended scope (toward nonpolar solutes) and tunable selectivity. PMID- 12553772 TI - A sample introduction technique for obtaining a multiphoton ionization spectrum using picogram quantities of analyte in supersonic jet spectrometry. AB - A new sample introduction techniquefor ensuring the safety of a researcher during the measurement of hazardous compounds, such as dioxins, by supersonic jet spectrometry is described. The solution containing the analyte is introduced beneath the nozzle through a narrow capillary with a glass frit at the end, which is formed to vaporize only the solvent and to deposit the analyte. The first laser, that is, the second harmonic emission of a Nd: YAG laser, vaporizes the analyte and entrains it into a supersonic jet. The analyte molecule is ionized by the second laser, that is, an optical parametric oscillator. Ion measurement is achieved by a time-of-flight mass spectrometer. The analyte molecule is localized in the jet, thus permitting the amount of sample to be minimized. The minimum amount of phenol necessary for measurement of a multiphoton ionization spectrum by scanning the wavelength of the ionization laser in the 3-nm range was 180 pg, which is similar to or slightly less than the tolerable daily intake (TDI) of dioxin. PMID- 12553773 TI - Chemicofunctional membrane for integrated chemical processes on a microchip. AB - Here we report a design and synthesis of a chemically functional polymer membrane by an interfacial polycondensation reaction and multilayer flow inside a microchannel. Single and parallel dual-membrane structures are successfully prepared by using organic/aqueous two-layer flow and organic/aqueous/organic three-layer flow inside the microchannel followed by an interfacial polycondensation reaction. By using the inner-channel membrane, permeation of ammonia species through the inner-channel membrane is successfully achieved. Furthermore, horseradish peroxidase is immobilized on one side of the membrane surface to integrate the chemical transform function onto the inner-channel membrane. Here substrate permeation through the membrane and subsequent chemical transformation at the membrane surface are realized. The polymer membrane prepared inside the microchannel has an important role in ensuring stable contact of different phases such as gas/liquid or liquid/ liquid and the permeation of chemical species through the membrane. Furthermore, membrane surface modification chemistry allows chemical transformation of permeated chemical species. These methods are expected to lead to development of complicated and sophisticated chemical systems involving membrane permeation and chemical reactions. PMID- 12553774 TI - Micro X-ray fluorescence imaging without scans: toward an element-selective movie. AB - Micro X-ray fluorescence imaging is a promising method for obtaining positional distribution on specific elements in a nondestructive manner. So far, the technique has usually been performed by a 2D positional scan of a sample against a collimated beam. However, the total measuring time can become quite long, since a number of scanning points are needed in order to obtain a high-quality image. The present report discusses a completely different way of performing imaging of elements much more quickly. A combination of grazing-incidence geometry using a rather wide beam and parallel optics for detecting X-rays can produce an X-ray fluorescence image with approximately 1 M pixels and with approximately 20-microm resolution in 1-2 min or less. The technique has the potential to open up new frontiers in X-ray imaging, particularly in element-selective movie applications. PMID- 12553775 TI - Public preferences for an attribution to government or to medical research versus unattributed messages in cigarette warning labels in Israel. AB - The history of cigarette warning labels is fraught with dispute, and in many instances official anti-smoking warnings that appear on cigarette packets are the result of political compromise. Despite mixed findings on the effectiveness of these warning labels, they are viewed as a cost-effective anti-smoking measure by their mere presence and as an important part of larger anti-smoking efforts. Israel's Ministry of Health, in its recent initiative to introduce a new series of warning labels, has grappled with the attribution issue. A committee established by the Ministry to revise the warnings decided to survey public opinion to guide its decision regarding to whom warnings should be attributed and to counter tobacco lobby oppositions. Two surveys were conducted: a limited phone survey of the adult population (n = 1000) and a face-to-face survey of 200 adult smokers. Findings indicate there was little support for unattributed warnings and that smokers, when presented with actual warnings, tended to favor the attribution to "medical studies." Nonsmokers were somewhat more likely to prefer an attribution to the Ministry of Health, explaining that it is "responsible for the topic" or "has the authority." Attributional preferences were found to be associated to some extent with educational level. Discrepancies found between preferences of light smokers across the 2 surveys suggest that the method of preference elicitation may play an important role. The discussion of the attributional preferences draws from the persuasion literature and it is suggested that warning messages should be matched with selected sources. PMID- 12553776 TI - An interactional structure of medical activities during acute visits and its implications for patients' participation. AB - Within the context of primary-care, physician-patient visits, researchers have documented both patients' low levels of communicative participation (e.g., question asking) and the advantages of such participation to healthcare (e.g., improved physical health and satisfaction). Prior research has offered a variety of partial, non-exclusive explanations for patients' low levels of participation. This article investigates one underdeveloped source of explanation: the organization of interaction itself. This article argues that the establishment of new medical problems in acute visits makes relevant an organized structure of social action that is composed of an ordered series of medical activities: establishing the reason for the visit, physicians gathering additional information (i.e., history taking and physical examination), physicians delivering diagnoses, and physicians providing treatment recommendations. This "project" of medical activity shapes physicians' and patients' understanding and production of communicative behavior. Using the method of conversation analysis, and analyzing transcribed audio- and videotape data of actual acute visits, this article describes and grounds this project and discusses its implications for research, theory, and improvement on patient participation. PMID- 12553777 TI - Informatics tools and medical communication: patient perspectives of "knowledge coupling" in primary care. AB - The use of various "informatics tools" for routine patient care promises to radically alter the ways whereby medical knowledge and information are processed and applied. In so doing, it can also change the nature of the information exchange and relational communication that occurs between patients and caregivers. This investigation examines how patients view "knowledge coupling" (KC) tools as they are routinely used in a "coupler-centered" family practice (CCP). How do patients view these tools as influencing information exchange and their relationship with caregivers? Based upon close- and open-ended questions of a random sample of patients in the CCP, this study found that most respondents viewed KC informatics tools as elevating the information exchange that occurs between patients and caregivers-increasing the extent to which patients are "heard and understood" by caregivers and come to "know and understand" what is ailing them. Many respondents report being empowered by the use of the tools, and having greater confidence in the care and advice caregivers offer. A few others, however, suggest diminished confidence in care and a more "impersonal" environment, resulting specifically from the use of the computer in the office. The article concludes by suggesting ways whereby such deficiencies might be remedied. PMID- 12553778 TI - Examining influences of pharmacists' communication with consumers about antibiotics. AB - A national study was conducted of community pharmacists to examine influences on their communication with consumers about antibiotics and antibiotic resistance. The objective was twofold: to explore the nature of barriers that might be changed to allow pharmacists greater participation in educational campaigns, and to discover which variables would best predict communication using a modification of the Theory of Reasoned Action model. Whereas most pharmacists agreed that their role in educating patients is important, they also noted several barriers prevent them from engaging in campaigns, such as time constraints, lack of educational materials, and fear of harming relations with physicians. Pharmacists' discussion about antibiotics in general was predicted primarily by attitudes about their role and efficacy, by their autonomy, and by enabling measures-such as pamphlets-that would better aid them in participating in a judicious antibiotic use campaign. Discussion about resistance was predicted by attitudes and enabling measures. PMID- 12553779 TI - The linear interaction model of personality effects in health communication. AB - The recent growth of research in message tailoring has opened up new avenues for researchers to use personality variables for message delivery. This article builds on research on idiocentrism and self-monitoring to propose a framework for message appeal construction. Based on a scheme for appeal categorization borrowed from commercial marketing, the article suggests that low and high idiocentrics differ from each other in the way they respond to appeal types. Similarly, significant differences are demonstrated between low and high self-monitors in the realm of their response to message appeals. A linear interaction model is proposed to document the combined effects of self-monitoring and idiocentrism. PMID- 12553780 TI - Diastereo- and enantioselective catalytic carbometallative aldol cycloreduction: tandem conjugate addition-aldol cyclization. AB - A catalytic diastereo- and enantioselective method for tandem conjugate addition aldol cyclization is described. This methodology enables the formation of five- and six-membered ring products from aromatic and aliphatic mono-enone mono-ketone precursors. Notably, in a single manipulation, three contiguous stereogenic centers are created with high levels of relative and absolute stereocontrol. PMID- 12553781 TI - 3'-Ferrocene-labeled oligonucleotide chains end-tethered to gold electrode surfaces: novel model systems for exploring flexibility of short DNA using cyclic voltammetry. AB - The flexibility of DNA is of central importance in biology, medicine, materials science, and mechanical engineering. In this study, we report an unprecedented electrochemical approach for investigating the flexibility of a short (typically 20-base), surface end-tethered single-stranded synthetic DNA oligonucleotide and of its postformed DNA duplex, taking as an example the homopolymer (dT)20 sequence in the regime of very high ionic strength ( approximately 1 M). PMID- 12553782 TI - Differential receptors create patterns diagnostic for ATP and GTP. AB - Herein we report the combination of a library of resin-bound sensors along with a multicomponent sensor array. This novel combinatorial array sensor system shows selectivity for nucleotide phosphates in solution. The design of the anchored receptor includes a 1,3,5-trisubstituted-2,4,6-triethylbenzene scaffold coupled with peptide libraries. Each chemosensor is placed into a micromachined cavity within a silicon wafer, and the optical changes observed by a charged-coupled device result in near-real-time digital analysis of solutions. A colorimetric displacement assay was performed, and time-dependent imaging studies of the selected sensing ensembles result in a differential responses upon addition of adenosine 5'-triphosphate (ATP), adenosine 5'-monophosphate (AMP), or guanosine 5'-triphosphate (GTP). An advantage to this approach is that it creates an array of sensors that gives a fingerprint response for each analyte. Principal component analysis indicates that the library of chemosensors can differentiate between ATP, GTP, and AMP. On the basis of factor loading values, individual sensors from the library were sequenced to elucidate their chemical composition. PMID- 12553783 TI - A potent, water-soluble and photoinducible DNA cross-linking agent. AB - Water-soluble DNA cross-linking phenol and biphenol derivatives 3 and 6 have been synthesized by a Mannich reaction. Both of them can cross-link DNA by photoactivation using visible light (wavelength > 400 nm). Compound 6 can cross link DNA at pH 5.0 and 7.7, whereas no cross-link was observed at pH 10.0. Density functional theory (DFT) calculation indicated that 6 displays a twist structure. Therefore, it could bind to DNA naturally and has potent property to cross-link DNA after photoactivation. PMID- 12553784 TI - Photoinduced electron injection from Ru(dcbpy)2(NCS)2 to SnO2 and TiO2 nanocrystalline films. AB - Photoinduced electron injection from the sensitizer Ru(dcbpy)2(NCS)2 (RuN3) into SnO2 and TiO2 nanocrystalline films occurs by two distinct channels on the femto- and picosecond time scales. The faster electron injection into the conduction band of the different semiconductors originates from the initially excited singlet state of RuN3, and occurs in competition with intersystem crossing. The rate of singlet electron injection is faster to TiO2 (1/55 fs-1) than to SnO2 (1/145 fs-1), in agreement with higher density of conduction band acceptor states in the former semiconductor. As a result of competition between the ultrafast processes, for TiO2 singlet, whereas for SnO2 triplet electron injection is dominant. Electron injection from the triplet state is nonexponential and can be fitted with time constants ranging from approximately 1 ps (2.5 ps for SnO2) to approximately 50 ps for both semiconductors. The major part of triplet injection is independent of the semiconductor and is most likely controlled by intramolecular dynamics in RuN3. The overall time scale and the yield of electron injection to the two semiconductors are very similar, suggesting that processes other than electron injection are responsible for the difference in efficiencies of solar cells made of these materials. PMID- 12553785 TI - To fold or to assemble? AB - We introduce a new class of foldable oligomers consisting of alternating rigid and flexible regions. The rigid segments overlap to give pi-stacked folded conformers whose formation is driven mostly by pi-pi molecular orbital overlaps. As the oligomer concentration increases, the folded molecular structures further self-assemble into larger nanostructures. The dynamic processes of folding and self-organization are monitored with absorption, fluorescence, and NMR spectroscopies. Our results show that folding dominates at low concentrations (< approximately 1 mM) and precedes self-assembly, which occurs over the initial concentration range of approximately 1-100 mM. PMID- 12553786 TI - Novel self-assembled monolayers of dipolar ruthenium(III/II) pentaammine(4,4' bipyridinium) complexes on ultrathin platinum films as redox molecular switches. AB - We report the first example of a reversible redox-molecular switch based on dipolar ruthenium(III/II) pentaammine(4,4'-bipyridinium) complexes self-assembled on an optically transparent metal electrode, whose change in molecular properties can be simply detected by transmission spectroscopy with a conventional spectrophotometer. PMID- 12553787 TI - Nerve agents detection using a Cu2+/L-cysteine bilayer-coated microcantilever. AB - A Cu2+/l-cysteine bilayer-coated microcantilever undergoes bending upon exposure to dimethyl methyl phosphonate at concentration as low as 10-15 M due to the complexation of a phosphonyl group with the Cu2+/l-cysteine bilayer on the microcantilever surface. PMID- 12553788 TI - Selectivity of calix[4]arene-bis(benzocrown-6) in the complexation and transport of francium ion. AB - It is shown for the first time that a representative member of the "cesium selective" calix[4]arene-crown-6 family exhibits a high affinity for Fr+ ion. In the investigation, the transport of 221Fr+ and Cs+ ions by calix[4]arene bis(benzocrown-6) from an aqueous sodium nitrate solution into the water immiscible diluent 1,2-dichloroethane was measured and compared to address the question of selectivity of the calix-crown-6 cavity toward alkali metal ions of increasing size. Selective separation of 221Fr+ from its parent 225Ac and from the matrix Na+ ions was demonstrated. Higher distribution ratios were obtained than those for the Cs+ ion. The extraction equilibria were determined for the case of the Cs+ ion, and the same equilibria were shown to be applicable to the case of Fr+ with inclusion of additional competitive effects. PMID- 12553789 TI - A highly stable N-heterocyclic carbene complex of trichloro-oxo-vanadium(V) displaying novel Cl-Ccarbene bonding interactions. AB - Reaction of 1,3-dimesitylimidazol-2-ylidene and trichloro-oxo-vanadium(V) yields an air stable 1:1 adduct, which demonstrates the utility of N-heterocyclic carbenes to stabilize metal complexes in high oxidation states. The molecular structure of this compound reveals that the chloride ligands cis to the carbene are oriented toward the Ccarbene atom. Density functional theory calculations show that a bonding interaction occurs between lone pairs of these chlorides and the formally unoccupied p-orbital of the carbene. Previous studies indicated that this orbital was not involved in the bonding of N-heterocyclic carbenes to transition metals. The observed interaction therefore represents a new bonding mode for these widely used ligands. PMID- 12553790 TI - Combinatorial approach to organelle-targeted fluorescent library based on the styryl scaffold. AB - The first fluorescent styryl dye library with a broad color range was synthesized by combinatorial condensation of various aldehydes and methyl pyridinium compounds, and their applications as organelle specific staining probes were demonstrated. PMID- 12553791 TI - Solvent polarity at an aqueous/alkane interface: the effect of solute identity. AB - Resonance-enhanced second harmonic generation (SHG) has been used to probe the solvatochromic behavior of two small, aromatic chromophores adsorbed to the aqueous/cyclohexane, liquid/liquid interface. SHG spectra of p-nitrophenol (PNP) and 2,6-dimethyl-PNP (dmPNP) indicate that these two chromophores sample markedly different environments. PNP sees a polar, waterlike environment, whereas solvent polarity surrounding dmPNP is dominated by the nonpolar, organic phase. Results suggest that subtle changes in solute structure can change the distribution of solutes across an interface and thus change a solute's local solvation environment. PMID- 12553792 TI - Ionicity of intramolecular charge-transfer molecule Ddelta+-pi-Adelta- based on 1,3,3-trimethyl-2-methyleneindoline and 7,7,8,8-tetracyanoquinodimethane. AB - We report a simple and convenient method to evaluate the relative ionicity delta of the typical intramolecular charge-transfer (CT) molecules linked by pi-bond Ddelta+-pi-Adelta- (D, donor; A, acceptor; delta, degree of CT) derived from 1,3,3-trimethyl-2-methylene indoline and TCNQ derivatives on the basis of the solvatochromic effects of the CT band in various solvents and their redox properties. The functionality of the intramolecular CT molecules as dyes, photovoltaic materials, nonlinear optics, rectifiers, and conductors is discussed in terms of delta. PMID- 12553793 TI - Hydrometal analogues of aromatic hydrocarbons: a new class of cyclic hydrocoppers(I). AB - A new class of cyclic hydrocoppers(I) with the general formula CunHn (n = 3-6), resembling the cyclic hydrocarbon analogues, were predicted by means of DFT calculations to be stable molecules adopting a perfect planar configuration of high-symmetry characteristic of the aromatic systems. PMID- 12553794 TI - Nonaqueous synthesis and selective crystallization of gallium sulfide clusters into three-dimensional photoluminescent superlattices. AB - A low-temperature, nonaqueous synthesis approach is described that produces a series of gallium sulfide and polysulfide three-dimensional superlattices from binary Ga-S and ternary Zn-Ga-S supertetrahedral clusters. The diversity of superlattices is achieved by modifying the cluster size, the cluster composition, and the inter-cluster linkage mode. Both pure T3 (Ga10S186-) (denoted as UCR 7GaS) and pure T4 (Zn4Ga16S3310-) (denoted as UCR-5ZnGaS) superlattices with ring sizes of 18 and 24 tetrahedral atoms have been made. Of particular interest is the synthesis of the T3-T4 hybrid superlattice (denoted as UCR-19) with an odd ring size of 21 tetrahedral atoms. Another unprecedented feature is the occurrence of the -S-S-S- polysulfide linkage between supertetrahedral clusters in UCR-18. The fluorescent emission wavelength of these materials ranges from 440 to 500 nm and fills the previously observed gap between open-framework oxides and indium sulfides. A comparative study shows that open-framework gallium sulfides are more thermally stable than indium sulfides. They can also undergo ion exchange. It is suggested here that supertetrahedral clusters of different types coexist in a solution and can be selectively crystallized out with a proper choice of structure-directing agents. PMID- 12553795 TI - Asymmetric catalysis of Nozaki-Hiyama allylation and methallylation with a new tridentate bis(oxazolinyl)carbazole ligand. AB - This work describes the development of a new tridentate ligand effective for the asymmetric catalysis of Nozaki-Hiyama allylation and methallylation. Various aldehydes were allylated or methallylated with good enantioselectivity (86-96%), and a key intermediate of calcitriol lactone synthesis was also obtained with excellent diastereoselectivity (97% de, 91%). The enantioselective reaction catalyzed by this Cr-ligand complex is applicable to a broad range of aldehydes and has great potential for natural product synthesis. Another remarkable feature of this ligand is the stability of the Cr-ligand complex which was recovered after the enantioselective reaction and recycled twice without diminishing the enantioselectivity and yield. PMID- 12553796 TI - A proton-doped calix[4]arene-based conducting polymer. AB - Segmented conducting polymers based upon a calix[4]arene scaffold are reported. The cone conformation creates a zigzag orientation of the polymer segments. Their acid-dependent conductivities are similar to the strong pH conductivity dependence of polyaniline which is said to be acid dopable. On the other hand, they have a segmented structure that imposes greater localization of the carriers. The conductivity of such a system can be considered to result from rapid self-exchange between discrete units. Hence, electron exchange between radical cations and p-diquinone salts produces the high conductivity of these polymers. PMID- 12553797 TI - Novel PI analogues selectively block activation of the pro-survival serine/threonine kinase Akt. AB - The synthesis from l-quebrachitol of a series of 3-deoxygenated ether lipid-type phosphatidylinositol (PI) analogues is reported, that selectively block activation of Akt and downstream substrates without affecting activation of the upstream kinase, PDK-1, or other kinases downstream of ras such as MAPK in H157 and H1703 lung cancer cells that have high levels of constitutively active Akt. The 2-hydroxyl in these compounds was deleted or alkylated with the intent to preclude metabolic degradation of these compounds by PI-specific phospholipase C (PI-PLC). PI analogues with phosphate linkers are more effective than those with carbonate linkers. Specific inhibition of Akt by these compounds validates ligand design targeted to the PH domains of crucial signaling proteins, thus providing a unique class of possible cancer therapeutics. PMID- 12553798 TI - Spectroscopic evidence for site specific chemistry at a unique iron site of the [4Fe-4S] cluster in ferredoxin:thioredoxin reductase. AB - Ferredoxin:thioredoxin reductase (FTR) catalyzes the reduction of the disulfide in thioredoxin in two one-electron steps using an active site comprising a [4Fe 4S] in close proximity to a redox active disulfide. Mossbauer spectroscopy has been used to investigate the ligation and electronic properties of the [4Fe-4S] cluster in as-prepared FTR which has the active-site disulfide intact and in the N-ethylmaleimide (NEM)-modified form which provides a stable analogue of the one electron-reduced heterodisulfide intermediate and has one of the cysteines of the active-site disulfide alkylated with NEM. The results reveal novel site-specific cluster chemistry involving weak interaction of the active-site disulfide with a unique Fe site of the [4Fe-4S]2+ cluster in the resting enzyme and cleavage of the active-site disulfide with concomitant coordination of one of the cysteines to yield a [4Fe-4S]3+ cluster with a five-coordinate Fe site ligated by two cysteine residues in the NEM-modified enzyme. The results provide molecular-level insight into the catalytic mechanism of FTR and other Fe-S-cluster-containing disulfide reductases, and suggest a possible mechanism for the reductive cleavage of S-adenosylmethionine by the radical SAM family of Fe-S enzymes. PMID- 12553799 TI - Label-free chiral detection of melittin binding to a membrane. AB - The study presented here describes an innovative approach for the detection of surface-confined proteins using chiral second harmonic generation (C-SHG). A unique optical geometry has been employed which allows for the separation of the chiral and achiral nonlinear response. By utilizing this optical arrangement, the detection of chirality originating from melittin adsorbed to a planar supported lipid bilayer has been performed for the first time by C-SHG. Melittin binding to the membrane was monitored as a function of bulk concentration through detection of the C-SHG signal. Analysis of the C-SHG adsorption isotherms reveals Frumkin adsorption behavior with a positive interaction energy. The binding constant (Ka) obtained was determined to be (8.3 +/- 1.0) x 105 M-1. The results of these studies have far-reaching implication in the use of C-SHG for the label-free detection of protein association to surfaces and in the analysis of protein interfacial phenomena. PMID- 12553800 TI - Supramolecular H-bonded assemblies of redox-active metallodendrimers and positive and unusual dendritic effects on the recognition of H2PO4-. AB - The DSM polyamine dendrimers dend-DAB-(NH2)x of generations 1 (x = 4) to 4 (x = 32) form H-bonded dendritic assemblies with the phenol AB3 units p HOC6H4C(CH2CHCH2)3 and p-HOC6H4C{(CH2)3SiCH2NHCOFc}3 (Fc = ferrocenyl), as shown by the shifts of the NH2 and OH signals giving a concentration-dependent common signal between 2.4 and 4.1 ppm in CDCl3. The supramolecular dendrimers efficiently recognize H2PO4- anions with positive and unusual dendritic effects upon electrochemical titration involving half-stoichiometry for G1, a sudden cyclovoltammetry wave change at the equivalent point, and a dramatic intensity decrease of the new wave. PMID- 12553801 TI - Novel Cu(I) dinuclear complexes containing mu2-eta(2),eta(2)-type benzoquinone ligand. AB - Novel Cu(I) complexes containing a mu2-eta2,eta2-type benzoquinone ligand have been synthesized and crystallographycally characterized. These complexes are synthesized by the redox reaction of a copper-quinonoid pair and may indicate a significant intermediate in the Cu-BQ catalytic system. Moreover, this study is regarded as one of the good examples in the Cu(I)-olefin family of enhanced pi back-donation. PMID- 12553802 TI - A method for making inorganic and hybrid (organic/inorganic) fibers and vesicles with diameters in the submicrometer and micrometer range via sol-gel chemistry and electrically forced liquid jets. AB - Electrically driven liquid jets are combined with sol-gel methods to design vesicles and fibers made from inorganic oxides and hybrid materials with diameters in the micrometer and submicrometer range. The proposed materials synthesis method benefits greatly from the maturity of sol-gel chemistry and the generalities of a structure-directing phenomenon that is physical in nature. PMID- 12553803 TI - Solvent-free functionalization of carbon nanotubes. AB - A fundamentally new single-walled and multiwalled carbon nanotube sidewall functionalization technique has been developed in which solvent is not required and the reaction times are greatly shortened (1 h at 60 degrees C). Exploiting the long linear dimension of the nanotube ropes by macroscopic mechanical deformation, reactive sites are generated merely by mechanically deforming the tubes using a stir bar. This approach eliminates the need for large volumes of solvent ( approximately 2 L/g), which were formerly considered essential due to the insolubility of carbon nanotubes. Using a series of 4-substituted anilines and a nitrite, the aryl diazonium intermediates were generated in situ and permitted to react with the tubes. Raman, IR, and UV spectroscopies, coupled with thermogravimetric analyses and solubility studies, support the assignments. PMID- 12553804 TI - Ultratrace kinetic measurements of the reduction of methylene blue. AB - The kinetics of methylene blue reduction by ascorbic acid in acetonitrile was investigated by cavity ring-down spectroscopy. Because of our high sensitivity we were able to use very low concentrations (1-10 nM) of the dye. Under these conditions, we observed a second-order loss of dye as well as a competing back reaction with dissolved oxygen. The use of an inexpensive diode laser and a relatively simple setup should make ultratrace kinetic studies more accessible. PMID- 12553805 TI - Reaction of elemental sulfur with a copper(I) complex forming a trans-mu-1,2 end on disulfide complex: new directions in copper-sulfur chemistry. AB - Elemental sulfur (S8) was found to react with [(TMPA)CuI(CH3CN)]+ to form the trans-mu-1,2 end-on disulfide complex [(TMPA)Cu-S-S-Cu(TMPA)]2+. The X-ray structure of this centrosymmetric disulfide complex shows a Cu(1)-S(1) bond length of 2.280(2) A and a S(1)-S(1A) bond length of 2.044(4) A. PMID- 12553806 TI - Aerobic oxidations catalyzed by chromium corroles. AB - Oxochromium(V) complexes of 5,10,15-tris(pentafluorophenyl)corrole and brominated derivatives oxygenate substrates (triphenylphosphine and norbornene) with concomitant production of chromium(III). Regeneration of CrVO by reaction of dioxygen with CrIII completes an aerobic catalytic cycle, with very large solvent effects; in acetonitrile, rapid initial turnovers observed initially are shut down by formation of CrIVO, while in toluene, THF, and methanol, relatively slow reactions are further inhibited by product formation. PMID- 12553807 TI - Excited-state intramolecular proton transfer in o-hydroxybiaryls: a new route to dihydroaromatic compounds. AB - An excited-state intramolecular proton transfer (ESIPT) from the phenol OH to the 7'-carbon on the naphthyl ring in o-(1-naphthyl)phenol (3) and 1-(1'-naphthyl)-2 naphthol (4) leads to efficient (Phi = 0.1-0.2) formation of the corresponding dihydrobenzoxanthenes (5 and 7) via quinone methide intermediates. This new reaction represents a clean, efficient, and high-yielding route to benzoxanthenes and dihydrobenzoxanthenes. A related ESIPT of similar efficiency has been detected at the 2'-aromatic position in these systems, by deuterium labeling studies. PMID- 12553808 TI - Gelation of ionic liquid-based electrolytes with silica nanoparticles for quasi solid-state dye-sensitized solar cells. AB - For the first time silica nanoparticles were used to solidify ionic liquids. These ionic liquid-based quasi-solid-state electrolytes were successfully employed for regenerative photoelectrochemical cells that yielded 7% efficiency at AM 1.5 sunlight in combination with an amphiphilic ruthenium polypyridyl photosensitizer. PMID- 12553809 TI - Optical determination of the single-ion zero-field splitting in large spin clusters. AB - The dodecametallic Cr(III) cluster has an S = 6 ground state with an axial zero field splitting (ZFS) of DS=6 = +0.088 cm-1. Analysis of high-resolution optical data (MCD) allows us to determine the single-ion ZFS of the constituent Cr(III) ions directly (D = -1.035 cm-1). A vector coupling analysis demonstrates that the cluster ZFS is almost entirely single-ion in origin. Thus, the relative orientations of the local and cluster magnetic axes can lead to cluster ZFS of opposite sign to the single-ion even when this is the only significant contribution. PMID- 12553810 TI - Remarkable metal-rich ternary chalcogenides Sc14M3Te8 (M = Ru, Os). AB - In this novel motif, scandium atoms define infinite parallel chains of alternate trans-face-sharing cubes and pairs of square antiprisms in which each polyhedron is also centered by an M atom (M = Ru, Os). These chains are further linked into a three-dimensional structure by Sc(Te2Te4/2) octahedra. Physical property measurements show Sc14Ru3Te8 to be metallic and Pauli-paramagnetic, consistent with the results of extended Huckel band structure calculations. Matrix effects are evident in the dimensions within the chains. The major interactions are Sc-M and Sc-Te. PMID- 12553811 TI - 15N NMR study on site-selective binding of metal ions to guanine runs in DNA: a good correlation with HOMO distribution. AB - 15N NMR of DNA containing 15N-N7-enriched guanine (G) in the presence of paramagnetic ions (Mn(II) and Co(II)) was investigated. As the concentration of metal ion was increased, 15N NMR signals of the 5'G of GG and the middle G of GGG broadened site-selectively, indicating that electron-donating sites in G runs preferentially localize on the 5'G of GG and the middle G of GGG. The selectivity for G-metal ion interaction observed in this study was in good agreement with calculated HOMO distribution of G runs. PMID- 12553812 TI - Novel fluorophores for single-molecule imaging. AB - Nonlinear optical chromophores based on dicyanodihydrofuran acceptors paired with amine donors have been found to exhibit sufficiently large fluorescence quantum yields and stability to enable single-molecule detection in polymeric hosts. To illustrate the breadth of this class, six fluorophores are presented, spanning the emission range from 505 to 646 nm. In contrast to conventional single molecule fluorophores, the new molecules feature sensitivity to local rigidity, large ground-state dipole moments, and large polarizability anisotropies, properties that can be used to design new reporter experiments at the single molecule level. PMID- 12553813 TI - A new form of kinetic isotope effect. Dynamic effects on isotopic selectivity and regioselectivity. AB - The intramolecular H/D kinetic isotope effect in the ene reaction of singlet oxygen with tetramethylethylene is studied using quasiclassical direct dynamics calculations on a B3LYP/6-31G* potential energy surface. Starting from the area of the energy surface around a valley-ridge inflection point, random trajectories lead to predominantly H abstraction over D abstraction, despite the symmetry of the surface and the absence of a barrier to either reaction. This demonstrates a new form of kinetic isotope effect, unrelated to the usual effect of zero-point energies on barriers. Dynamics calculations on the reaction of cis-2-pentene predict the experimentally observed mixture of regioisomeric products, while the minimum-energy path leads to only one product. For energy surfaces containing two adjacent saddle points, dynamics effects are important for understanding both product and isotopic selectivity, and this should be considered in the interpretation of experimental results. PMID- 12553814 TI - p-Benzosemiquinone radical anion on silver nanoparticles in water. AB - This communication reports the SERS observation of p-benzosemiquinone radical anion, produced on reduction of p-benzoquinone by Ag nanoparticles at the metal water interface. The species is positively identified by comparison of the SERS spectrum with the resonance Raman spectra of the radical anion in aqueous solution. This is a rare SERS observation of a radical intermediate formed by surface reaction on nanosize silver particles in solution. PMID- 12553815 TI - Stereochemistry of palladium-mediated synthesis of PAMP-BH3: retention of configuration at P in formation of Pd-P and P-C bonds. AB - Treatment of Pd((S,S)-Chiraphos)(o-An)(I) (3, o-An = o-MeOC6H4) with either enantiomer of highly enantioenriched PH(Me)(Ph)(BH3) (1) gave the phosphido borane complex Pd((S,S)-Chiraphos)(o-An)(P(Me)(Ph)(BH3)) (4) with retention of configuration at phosphorus, as shown by X-ray crystal structure determinations for both diastereomers of 4. Heating either diastereomer of 4 with diphenylacetylene gave Pd((S,S)-Chiraphos)(PhCCPh) (5) and P(o-An)(Me)(Ph)(BH3) (2) with retention of configuration at phosphorus. PMID- 12553816 TI - Voltammetry and electron-transfer dynamics in a molecular melt of a 1.2 nm metal quantum dot. AB - New molecular melts of nanoparticles have been obtained by place exchanging thiolated poly(ethyleneglycol, MW = 350) ligands into the monolayer shells of the quantum dot nanoparticle Au38(phenylethylthiolate)24. These melts are nearly monodisperse in monolayer protected Au clusters with core diameters of approximately 1.2 nm. LiClO4 electrolyte can be dissolved in the melt via the PEG component of the protecting monolayer, producing an ionically conductive nanophase and enabling voltammetry of the undiluted, semisolid nanoparticle molecular melt. The optical and electrochemical charging properties of the small nanoparticles have molecule-like characteristics (as opposed to quantized double layer charging) both in dilute fluid-solvent solutions and as undiluted melts. Potential step chronoamperometry shows that electronic charge is transported through the melt by diffusion-like core-core electron hopping reactions with a rate constant of 2 x 104 s-1. PMID- 12553817 TI - Novel Z-selective head-to-head dimerization of terminal alkynes catalyzed by lanthanide half-metallocene complexes. AB - Various terminal alkynes have been cleanly dimerized into the corresponding head to-head (Z)-enynes by use of the half-metallocene lutetium alkyl complexes Me2Si(C5Me4)(NAr)Lu(CH2SiMe3)(THF) (Ar = Ph, C6H3Me2-2,6, C6H2Me3-2,4,6) as catalysts. Aromatic C-Cl, C-Br, and C-I bonds, which are known to be extremely susceptible to reductive cleavage by transition metals, survived in the present reactions. The corresponding dimeric alkynide species [Me2Si(C5Me4)(NAr)Lu(mu CCR)]2 are thought to be the true catalysts, some of which have been isolated and structurally characterized. These alkynide species were thermally stable and soluble at the reaction temperatures (80-110 degrees C), but they precipitated upon cooling to room temperature after completion of the reaction. Therefore, this catalyst system works homogeneously but can be separated and reused, thus constituting the first example of a recyclable catalyst system for the dimerization of terminal alkynes and also the first example of (Z)-selective head to-head dimerization of aromatic terminal alkynes. PMID- 12553818 TI - Highly negative homotropic allosteric binding of viologens in a double-cavity porphyrin. AB - The synthesis of a double-cavity porphyrin with interesting allosteric binding properties toward viologens (N,N'-disubstituted 4,4'-bipyridines) is described. The porphyrin host forms very strong 1:2 complexes with viologens, displaying a negative allosteric behavior. The first viologen guest binds exceptionally tight (K > 107 M-1), and the second guest binds much more weakly (DeltaDeltaG = 9-15 kJ mol-1). The allosteric effect, one of the highest reported so far, originates in structural changes upon binding the first ligand, closely following the sequential (or induced-fit) theory of allosteric interactions by Koshland, Nemethy, and Filmer (the KNF-model). PMID- 12553819 TI - Remarkably favorable hydration of carbonyl substituents in dicationic bis(arene) ruthenium complexes. AB - The addition of water and methanol to the carbonyl substituents in the dicationic bis(arene)ruthenium complexes [(p-cymene)Ru(C6H5CHO)][OTf]2 (1) and [(p cymene)Ru(C6H5C(O)CH3)][OTf]2 (2) is remarkably favorable-the equilibrium constants for these reactions are 1 million times greater than those for uncomplexed benzaldehyde and acetophenone. The addition of methanol to the carbonyl group in the o-anisaldehyde complex, [(p-cymene)Ru(C6H4{OCH3}CHO)][OTf]2 (6), occurs with a high degree of kinetic selectivity (94% de). PMID- 12553820 TI - Catalytic asymmetric silane alcoholysis: practical access to chiral silanes. AB - A new asymmetric synthesis of chiral silanes has been developed. Chiral phosphine modified copper complexes catalyze the alcoholysis of prochiral dihydrosilanes with good to excellent stereoselectivity at silicon. The application of this methodology to the tandem silylformylation-allylsilylation of alkynes has been demonstrated. PMID- 12553821 TI - The first enantioselective organocatalytic Mukaiyama-Michael reaction: a direct method for the synthesis of enantioenriched gamma-butenolide architecture. AB - The first enantioselective organocatalytic Mukaiyama-Michael reaction using alpha,beta-unsaturated aldehydes has been accomplished. The use of iminium catalysis has provided a new strategy for the enantioselective addition of 2 silyloxy furans to unsaturated aldehydes to generate a variety of butenolide systems, an important chiral synthon found among many natural isolates. The (2S,5S)-5-benzyl-2-tert-butyl-imidazolidinone amine catalyst has been found to mediate the conjugate addition of a wide variety of substituted and unsubstituted silyloxy furans to unsaturated aldehydes. A diverse range of aldehyde substrates can be accommodated in this new organocatalytic transformation. Application of this new asymmetric technology to the enantioselective total synthesis of spiculisporic acid and the corresponding 5-epi-spiculisporic acid analogue is also discussed. PMID- 12553822 TI - Sequence specific fluorescence detection of double strand DNA. AB - Methods for the fluorescent detection of specific sequences of double strand DNA in homogeneous solution may be useful in the field of human genetics. A series of hairpin polyamides with tetramethyl rhodamine (TMR) attached to an internal pyrrole ring were synthesized, and the fluorescence properties of the polyamide fluorophore conjugates in the presence and absence of duplex DNA were examined. We observe weak TMR fluorescence in the absence of DNA. Addition of >/=1:1 match DNA affords a significant fluorescence increase over equimolar mismatch DNA for each polyamide-TMR conjugate. Polyamide-fluorophore conjugates offer a new class of sensors for the detection of specific DNA sequences without the need for denaturation. The polyamide-dye fluorescence-based method can be used to screen in parallel the interactions between aromatic ring pairs and the minor groove of DNA even when the binding site contains a non-Watson-Crick DNA base pair. A ranking of the specificity of three polyamide ring pairs-Py/Py, Im/Py, and Im/Im was established for all 16 possible base pairs of A, T, G, and C in the minor groove. We find that Im/Im is an energetically favorable ring pair for minor groove recognition of the T.G base pair. PMID- 12553823 TI - Dual wavelength parametric test of two-state models for circular dichroism spectra of helical polypeptides: anomalous dichroic properties of alanine-rich peptides. AB - A two-state helix-coil model underlies all calculations of fractional helicities FH from CD spectra of helical polypeptides. The presence of an isodichroic point near 203 nm is widely assumed to validate this model, but is shown here to provide inadequate validation for alanine-rich peptides. A parametric correlation with constant slope B between CD ellipticities at a pair of wavelengths is introduced as a more rigorous two-state test. Correlations of temperature dependent [theta](222) vs [theta](208) values are reported for a variety of peptides. Constant slopes B are observed for literature CD data obtained from fragments of helical proteins and dimeric helical coiled coils, but parametric correlations of CD data for alanine-rich peptides consistently exhibit anomalous concave upward curvature, characterized by local slopes that are linearly temperature dependent. Low-temperature CD studies together with parametric correlations at a series of wavelengths demonstrate that the curvature anomaly is maximal at 222 nm and localized in the 215-230 nm wavelength region. Precedented structural variation of the phi, psi dihedral angles of the alpha-helix is suggested as a possible explanation. For the important case of alanine-rich peptides, experiments are proposed that may yield temperature corrections for [theta](222) and permit reliable calculations of FH from [theta](222) values. PMID- 12553824 TI - Impact of azaproline on amide cis-trans isomerism: conformational analyses and NMR studies of model peptides including TRH analogues. AB - The beta-turn is a well-studied motif in both proteins and peptides. Four residues, making almost a complete 180 degree-turn in the direction of the peptide chain, define the beta-turn. Several types of the beta-turn are defined according to Phi and Psi torsional angles of the backbone for residues i + 1 and i + 2. One special type of beta-turn, the type VI-turn, usually contains a proline with a cis-amide bond at residue i + 2. In an aza-amino acid, the alpha carbon of the amino acid is changed to nitrogen. Peptides containing azaproline (azPro) have been shown to prefer the type VI beta-turn both in crystals and in organic solvents by NMR studies. MC/MD simulations using the GB/SA solvation model for water explored the conformational preferences of azPro-containing peptides in aqueous systems. An increase in the conformational preference for the cis-amide conformer of azPro was clearly seen, but the increased stability was relatively minor with respect to the trans-conformer as compared to previous suggestions. To test the validity of the calculations in view of the experimental data from crystal structures and NMR in organic solvents, [azPro(3)]-TRH and [Phe(2), azPro(3)]-TRH were synthesized, and their conformational preferences were determined by NMR in polar solvents as well as the impact of the azPro substitution on their biological activities. PMID- 12553825 TI - A new approach to mineralization of biocompatible hydrogel scaffolds: an efficient process toward 3-dimensional bonelike composites. AB - As a first step toward the design and fabrication of biomimetic bonelike composite materials, we have developed a template-driven nucleation and mineral growth process for the high-affinity integration of hydroxyapatite with a poly(2 hydroxyethyl methacrylate) (pHEMA) hydrogel scaffold. A mineralization technique was developed that exposes carboxylate groups on the surface of cross-linked pHEMA, promoting high-affinity nucleation and growth of calcium phosphate on the surface, along with extensive calcification of the hydrogel interior. Robust surface mineral layers a few microns thick were obtained. The same mineralization technique, when applied to a hydrogel that is less prone to surface hydrolysis, led to distinctly different mineralization patterns, in terms of both the extent of mineralization and the crystallinity of the apatite grown on the hydrogel surface. This template-driven mineralization technique provides an efficient approach toward bonelike composites with high mineral-hydrogel interfacial adhesion strength. PMID- 12553827 TI - Origin of the red shifts in the optical absorption bands of nonplanar tetraalkylporphyrins. AB - The view that the large red shifts seen in the UV-visible absorption bands of peripherally crowded nonplanar porphyrins are the result of nonplanar deformations of the macrocycle has recently been challenged by the suggestion that the red shifts arise from substituent-induced changes in the macrocycle bond lengths and bond angles, termed in-plane nuclear reorganization (IPNR). We have analyzed the contributions to the UV-visible band shifts in a series of nickel or zinc meso-tetraalkylporphyrins to establish the origins of the red shifts in these ruffled porphyrins. Structures were obtained using a molecular mechanics force field optimized for porphyrins, and the nonplanar deformations were quantified by using normal-coordinate structural decomposition (NSD). Transition energies were calculated by the INDO/S semiempirical method. These computational studies demonstrate conclusively that the large Soret band red shifts ( approximately 40 nm) seen for very nonplanar meso-tetra(tert-butyl)porphyrin compared to meso-tetra(methyl)porphyrin are primarily the result of nonplanar deformations and not IPNR. Strikingly, nonplanar deformations along the high frequency 2B(1u) and 3B(1u) normal coordinates of the macrocycle are shown to contribute significantly to the observed red shifts, even though these deformations are an order of magnitude smaller than the observed ruffling (1B(1u)) deformation. Other structural and electronic influences on the UV visible band shifts are discussed and problems with the recent studies are examined (e.g., the systematic underestimation of the 2B(1u) and 3B(1u) modes in artificially constrained porphyrin structures that leads to a mistaken attribution of the red shift to IPNR). The effect of nonplanar deformations on the UV-visible absorption bands is then probed experimentally with a series of novel bridled nickel chiroporphyrins. In these compounds, the substituent effect is essentially invariant and the amount of nonplanar deformation decreases as the length of the straps connecting adjacent meso-cyclopropyl substituents decreases (the opposite of the effect observed for conventional strapped porphyrins). Several spectroscopic markers for nonplanarity (UV-visible bands, resonance Raman lines, and (1)H NMR resonances) are found to correlate with time-averaged deformations obtained from an NSD analysis of molecular dynamics snapshot structures. These results suggest that UV-visible band shifts of tetrapyrroles in proteins are potentially useful indicators of changes in nonplanarity provided other structural and electronic factors can be eliminated. PMID- 12553826 TI - Heavy atom isotope effects on the reaction catalyzed by the oxalate decarboxylase from Bacillus subtilis. AB - Oxalate decarboxylase (OxDC) catalyzes a remarkable transformation in which the C C bond in oxalate is cleaved to give carbon dioxide and formate. Like the native OxDC isolated from Aspergillus niger, the recombinant, bacterial OxDC from Bacillus subtilis contains Mn(II) in its resting state and requires catalytic dioxygen for activity. The most likely mechanism for OxDC-catalyzed C-C bond cleavage involves the participation of free radical intermediates, although this hypothesis remains to be unequivocally demonstrated. Efforts to delineate the catalytic mechanism have been placed on a firm foundation by the high-resolution crystal structure of recombinant, wild type B. subtilis OxDC (Anand et al., Biochemistry 2002, 41, 7659-7669). We now report the results of heavy-atom kinetic isotope effect measurements for the OxDC-catalyzed decarboxylation of oxalate, in what appear to be the first detailed studies of the mechanism employed by OxDC. At pH 4.2, the OxDC-catalyzed formation of formate and CO(2) have normal (13)C isotope effects of 1.5% +/- 0.1% and 0.5% +/- 0.1%, respectively, while the (18)O isotope effect on the formation of formate is 1.1% +/- 0.2% normal. Similarly at pH 5.7, the production of formate and CO(2) exhibits normal (13)C isotope effects of 1.9% +/- 0.1% and 0.8% +/- 0.1%, respectively, and the (18)O isotope effect on the formation of formate is 1.0% +/ 0.2% normal. The (18)O isotope effect on the formation of CO(2), however, 0.7% +/- 0.2%, is inverse at pH 5.7. These results are consistent with a multistep model in which a reversible, proton-coupled, electron transfer from bound oxalate to the Mn-enzyme gives an oxalate radical, which decarboxylates to yield a formate radical anion. Subsequent reduction and protonation of this intermediate then gives formate. PMID- 12553828 TI - Functional nanocomposites prepared by self-assembly and polymerization of diacetylene surfactants and silicic acid. AB - Conjugated polymer/silica nanocomposites with hexagonal, cubic, or lamellar mesoscopic order were synthesized by self-assembly using polymerizable amphiphilic diacetylene molecules as both structure-directing agents and monomers. The self-assembly procedure is rapid and incorporates the organic monomers uniformly within a highly ordered, inorganic environment. By tailoring the size of the oligo(ethylene glycol) headgroup of the diacetylene-containing surfactant, we varied the resulting self-assembled mesophases of the composite material. The nanostructured inorganic host altered the diacetylene polymerization behavior, and the resulting nanocomposites show unique thermo-, mechano-, and solvatochromic properties. Polymerization of the incorporated surfactants resulted in polydiacetylene (PDA)/silica nanocomposites that were optically transparent and mechanically robust. Molecular modeling and quantum calculations and (13)C spin-lattice relaxation times (T(1)) of the PDA/silica nanocomposites indicated that the surfactant monomers can be uniformly organized into precise spatial arrangements prior to polymerization. Nanoindentation and gas transport experiments showed that these nanocomposite films have increased hardness and reduced permeability as compared to pure PDA. Our work demonstrates polymerizable surfactant/silica self-assembly to be an efficient, general approach to the formation of nanostructured conjugated polymers. The nanostructured inorganic framework serves to protect, stabilize, and orient the polymer, mediate its performance, and provide sufficient mechanical and chemical stability to enable integration of conjugated polymers into devices and microsystems. PMID- 12553829 TI - Detection and amplification of a small enantiomeric imbalance in alpha-amino acids by a helical poly(phenylacetylene) with crown ether pendants. AB - We have designed a novel stereoregular poly(phenylacetylene) bearing the bulky crown ether as the pendant (poly-1) for the amino acid binding site. The polymer forms a one-handed helix upon complexation with l-amino acid perchlorates, and the complexes exhibit an induced circular dichroism (ICD) with the same Cotton effect signs in the polymer backbone region through a significant cooperative interaction. Poly-1 is highly sensitive to the amino acid chirality and can detect an extremely small enantiomeric imbalance in alpha-amino acids (less than 0.005% enantiomeric excess of alanine, for example). PMID- 12553830 TI - Atomic properties of selected biomolecules: quantum topological atom types of carbon occurring in natural amino acids and derived molecules. AB - We seek to recover rigorous atom types from amino acid wave functions. The atom types emerge from a cluster analysis operating on a set of seven atomic properties, including kinetic energy, volume, population, and dipole, quadrupole, octupole, and hexadecapole moments. These properties are acquired by partitioning the molecular electron density into quantum topological atoms. Wave functions are generated at the B3LYP/6-311+G(2d,p)//HF/6-31G(d) level for a sensible conformation of each of the 20 naturally occurring amino acids and smaller derived molecules, which together constitute a data set of 57 molecules. From this set 213 unique quantum topological carbons are obtained, which are linked according to the similarity of their properties. After introducing a statistical separation criterion, our cluster analysis proposes two representations: a cruder one with 5 atom types and a finer one with 21 atom types. The immediate coordination of the central carbon plays a major role in labeling the atom types. PMID- 12553831 TI - Noninnocence of the ligand glyoxal-bis(2-mercaptoanil). The electronic structures of [Fe(gma)]2, [Fe(gma)(py)] x py, [Fe(gma)(CN)]1-/0, [Fe(gma)I], and [Fe(gma)(PR3)(n)] (n = 1, 2). Experimental and theoretical evidence for "excited state" coordination. AB - The electronic structure of the known iron complexes [Fe(gma)](2) (S(t) = 0) (1)(6) and [Fe(gma)(py)].py (S(t) = 1) (2)(7) where H(2)(gma) represents glyoxal bis(2-mercaptoanil) has been shown by X-ray crystallography, Mossbauer spectroscopy, and density functional theory calculations to be best described as ferric (S(Fe) = 3/2) complexes containing a coordinated open-shell pi radical trianion (gma(*))(3)(-) and not as previously reported(6,7) as ferrous species with a coordinated closed-shell dianion (gma)(2)(-). Compound 1 (or 2) can be oxidized by I(2) yielding [Fe(III)(gma)I] (S(t) = 1/2) (3). With cyanide anions, complex 1 forms the adduct [(n-Bu)(4)N][Fe(III)(gma(*))(CN)] (S(t) = 1) (4), which can be one-electron oxidized with iodine yielding the neutral species [Fe(III)(gma)(CN)] (S(t) = 1/2) (5). With phosphines complex 1 also forms adducts(7) of which [Fe(III)(gma(*))(P(n-propyl)(3))] (S(t) = 1) (6) has been isolated and characterized by X-ray crystallography. [Fe(II)(gma)(P(n propyl)(3))(2)] (S(t) = 0) (7) represents the only genuine ferrous species of the series. Density functional theory (DFT) calculations at the BP86 and B3LYP levels were applied to calculate the structural as well as the EPR and Mossbauer spectroscopic parameters of the title compounds as well as of the known complexes [Zn(gma)](0/)(-) and [Ni(gma)](0/)(-). Overall, the calculations give excellent agreement with the available spectroscopic information, thus lending support to the following electronic structure descriptions: The gma ligand features an unusually low lying LUMO, which readily accepts an electron to give (gma(*))(3)( ). The one-electron reduction of [Zn(gma)] and [Ni(gma)] is strictly ligand centered and differences in the physical properties of [Zn(gma(*))](-) and [Ni(gma(*))](-) are readily accounted for in terms of a model that features enhanced back-bonding from the metal to the gma LUMO in the case of [Ni(gma(*))]( ). In the case of [Fe(gma)(PH(3))], [Fe(gma)(py)], and [Fe(gma)(CN)](-) an electron transfer from the iron to the gma LUMO takes place to give strong antiferromagnetic coupling between an intermediate spin Fe(III) (S(Fe) = 3/2) and (gma(*))(3)(-) (S(gma) = 1/2), yielding a total spin S(t) = 1. Broken symmetry DFT calculations take properly account of this experimentally calibrated electronic structure description. By contrast, the complexes [Fe(gma)(PH(3))(2)] and [Fe(PhBMA)] feature closed-shell ligands with a low-spin Fe(II) (S(Fe) = S(t) = 0) and an intermediate spin central Fe(II) (S(Fe) = S(t) = 1), respectively. The most interesting case is provided by the one-electron oxidized species [Fe(gma)(py)](+), [Fe(gma)I], and [Fe(gma)(CN)]. Here the combination of theory and experiment suggests the coupling of an intermediate spin Fe(III) (S(Fe) = 3/2) to the dianionic ligand (gma)(2)(-) formally in its first excited triplet state (S(gma) = 1) to give a resulting S(t) = 1/2. All physical properties are in accord with this interpretation. It is suggested that this unique "excited state" coordination is energetically driven by the strong antiferromagnetic exchange interaction between the metal and the ligand, which cannot occur for the closed shell form of the ligand. PMID- 12553832 TI - Hydrogen adsorption in nanoporous nickel(II) phosphates. AB - Hydrogen sorption in the nanoporous nickel phosphates VSB-1 and VSB-5 has been studied with a combination of BET, temperature programmed desorption (TPD), and inelastic neutron scattering (INS) measurements. H(2) BET isotherms for VSB-1 are similar to those seen in nanoporous zeolites, while VSB-5 adsorbs substantially more hydrogen due to a steep initial uptake at low partial pressures. TPD data show that hydrogen interacts strongly with VSB-5, with desorption peaks at 109 and 149 K in a nitrogen flow, whereas the absence of similar peaks for VSB-1 suggests a weaker interaction. INS spectra of the rotational tunnel transition of the adsorbed H(2) also reveal a strong interaction with the VSB-5 host. These data strongly suggest the existence of coordinatively unsaturated Ni(2+) sites accessible to H(2) molecules in the pores of VSB-5. PMID- 12553833 TI - Structure-function relations in self-assembled C18- and C20-sphingosines monolayers at gas/water interfaces. AB - Synchrotron X-ray studies and surface pressure versus molecular area (pi-A) isotherms of C18- and C20-sphingosines spread at air/water interfaces reveal unique interfacial properties with considerable differences between the two single hydrocarbon chain amino-alcohols. C20-sphingosine forms a crystalline monolayer with structural characteristics that are dominated by hydrogen bonding in the headgroup (common to its sphingolipid derivatives), whereas its natural counterpart C18-sphingosine forms a disordered liquidlike metastable monolayer and has to be spread in excess with a floating reservoir on the water surface to compensate for the high dissolution rate of molecules into the water subphase. The marginal affinity of C18-sphingosine to reside at the interface, the microcrystallization at very low densities, the corrugated monolayers it forms, and the strong interaction with the water surface are consistent with the roles that sphingolipids play in the life cycle of eukaryotic cells and as the building blocks of specialized membranes. PMID- 12553834 TI - Mechanism of ene reactions of singlet oxygen. A two-step no-intermediate mechanism. AB - The mechanism of the ene reaction of singlet ((1)delta(g)) oxygen with simple alkenes is investigated by a combination of experimental isotope effects and several levels of theoretical calculations. For the reaction of 2,4-dimethyl-3 isopropyl-2-pentene, the olefinic carbons exhibit small and nearly equal (13)C isotope effects of 1.005-1.007, while the reacting methyl groups exhibit (13)C isotope effects near unity. In a novel experiment, the (13)C composition of the product is analyzed to determine the intramolecular (13)C isotope effects in the ene reaction of tetramethylethylene. The new (13)C and literature (2)H isotope effects are then used to evaluate the accuracy of theoretical calculations. RHF, CASSCF(10e, 8o), and restricted and unrestricted B3LYP calculations are each applied to the ene reaction with tetramethylethylene. Each predicts a different mechanism, but none leads to reasonable predictions of the experimental isotope effects. It is concluded that none of these calculations accurately describe the reaction. A more successful approach was to use high-level, up to CCSD(T), single point energy calculations on a grid of B3LYP geometries. The resulting energy surface is supported by its accurate predictions of the intermolecular (13)C and (2)H isotope effects and a very good prediction of the reaction barrier. This CCSD(T)//B3LYP surface features two adjacent transition states without an intervening intermediate. This is the first experimentally supported example of such a surface and the first example of a valley-ridge inflection with significant chemical consequences. PMID- 12553835 TI - Shock-wave resistance of WS2 nanotubes. AB - The shock-wave resistance of WS(2) nanotubes has been studied and compared to that of carbon nanotubes. Detailed structural features of post-shock samples were investigated using HRTEM, XRD, and Raman spectroscopy. WS(2) nanotubes are capable of withstanding shear stress caused by shock waves of up to 21 GPa, although some nanotube tips and nanoparticles containing multiple structural defects in the bending regions are destroyed. Small WS(2) species, consisting of only a few layers, are extruded from the nanotubes. Well-crystallized tube bodies were found to exhibit significant stability under shock, indicating high tensile strength. XRD and Raman analyses have confirmed this structural stability. Under similar shock conditions, WS(2) tubes are more stable than carbon nanotubes, the latter being transformed into a diamond phase. WS(2) nanotubes containing small concentrations of defects possess significantly higher mechanical strength, and, as a consequence, hollow WS(2) nanoparticles are expected to act as excellent lubricants under much higher loading than was previously thought. PMID- 12553836 TI - Mechanistic studies on the interaction of reduced cobalamin (vitamin B12r) with nitroprusside. AB - The electron-transfer reaction between reduced cobalamin (Cbl(II)) and sodium pentacyanonitrosylferrate(II) (sodium nitroprusside, NP), as well as the subsequent processes following the electron-transfer step, were investigated by spectroscopic (UV-vis, (1)H NMR, EPR), electrochemical (CV, DPV) and kinetic (stopped-flow) techniques. In an effort to clarify the complex reaction pattern observed at physiological pH, systematic spectroscopic and kinetic studies were undertaken as a function of pH (1.8-9) and NP concentration (0.0001 - 0.09 M). The kinetics of the electron-transfer reaction was studied under pseudo-first order conditions with respect to NP. The reaction occurs in two parallel paths of different order, viz. pseudo-first and pseudo-second order with respect to the NP concentration, respectively. The contribution of each path depends on pH and the [NP]/[Cbl(II)] ratio. At low pH and total NP concentration (pH < 3, [NP]/[Cbl(II)] approximately 1), the cyano-bridged successor complex [Cbl(III) (mu-NC)-Fe(I)(CN)(3)(NO(+))](-) (1(s)()) is the final reaction product formed in an inner-sphere electron transfer reaction that is coupled to the release of cyanide from coordinated nitroprusside. At higher pH, subsequent reactions were observed which involve the attack of cyanide released in the electron transfer step on the initially formed cyano-bridged species, and lead to the formation of Cbl(III)CN and [Fe(I)(CN)(4)(NO(+))](2)(-). The strong dependence of the rate and mechanism of the subsequent reactions on pH is attributed to the large variation in the effective nucleophilicity of the cyanide ligand in the studied pH range. An alternative electron-transfer pathway observed in the presence of excess NP involves the reaction of the precursor complex [Cbl(II)-(mu-NC) Fe(II)(CN)(4)(NO(+))](2)(-) (1(p)()) with NP to give [Cbl(III)-(mu-NC) Fe(II)(CN)(4)(NO(+))](-) (2) and reduced nitroprusside, [Fe(CN)(5)NO](3)(-), as the initial reaction products. Analysis of the kinetic data allowed elucidation of the rate constants for the inner- and outer-sphere electron-transfer pathways. The main factors which influence the kinetics and thermodynamics of the observed electron-transfer steps are discussed on the basis of the spectroscopic, kinetic and electrochemical results. A general picture of the reaction pathways that occur on a short (s) and long (min to h) time scale as a function of pH and relative reactant concentrations is derived from the experimental data. In addition, the release of NO resulting from the one-electron reduction of NP by Cbl(II) was monitored with the use of a sensitive NO electrode. The results obtained in the present study are discussed in reference to the possible influence of cobalamin on the pharmacological action of nitroprusside. PMID- 12553837 TI - Electrodeposition of Au-Cd alloy nanostructures on Au(111). AB - This report concerns an in-situ scanning tunneling microscopy study of the initial stages in the formation of a Au-Cd alloy on the Au(111) herringbone reconstruction. Although Au-Cd nanoclusters of alloy have been observed in sulfate electrolyte by this group, alloy "nanowires" were observed to form preferentially in the hcp regions between the sets of "soliton" walls of the reconstruction only in the presence of chloride. The nanowires were formed at 0.55 V versus 3 M Ag/AgCl, corresponding to Cd underpotential deposition (upd). Upd is electrodeposition at a potential prior to that needed to deposit the bulk element. PMID- 12553838 TI - Entropic barriers in nanoscale adhesion studied by variable temperature chemical force microscopy. AB - Intermolecular interactions drive the vast majority of condensed phase phenomena from molecular recognition to protein folding to particle adhesion. Complex energy barriers encountered in these interactions include contributions from van der Waals forces, hydrogen bonding, and solvent medium. With the spectacular exception of hydrophobic interactions, contributions from the medium are usually considered secondary. We report a variable temperature force microscopy study of the interactions between several hydrogen bonds in different solvents that challenges this point of view. Surprisingly, we observed an increase in the strength of the interaction between carboxylic acid groups in ethanol as the temperature increased. Moreover, when we switched to a nonpolar solvent we observed the opposite behavior: The binding force decreased as the temperature increased. Kinetic model of bond dissociation provided quantitative interpretation of our measurements. We attributed the observed phenomena to a large entropic contribution from the ordered solvent layers that are forming on the probe and sample surfaces upon detachment. The observed reversal in the force vs temperature trend is a manifestation of a transition between thermodynamic and kinetic regimes of unbinding predicted by the model. Our results indicate that entropic barriers dominated by the interactions of solvent molecules with the surface exist in a much wider variety of systems than previously thought. PMID- 12553839 TI - Crown-annelated oligothiophenes as model compounds for molecular actuation. AB - Crown-annelated quater- (4T) and sexithiophenes (6T) with oligooxyethylene chains of various lengths attached at the 3-positions of the terminal thiophene rings have been synthesized. Analysis of the cation-binding properties of the macrocycles by (1)H NMR and UV-vis spectroscopy reveals the formation of a 1:1 complex with Ba(2+), Sr(2+), or Pb(2+) and shows that cation complexation results in a conformational transition in the pi-conjugated system. Theoretical analysis of this process by density functional methods predicts that this conformational transition results in a narrowing of the highest occupied-lowest unoccupied molecular orbital gap of the conjugated system with a decrease of the redox potentials (E(0)(1) and E(0)(2)) associated with the formation of the 4T cation radical and dication. Cyclic voltammetry shows that, depending on the binding constant, the presence of metal cation produces a negative or a positive shift of E(0)(1) while E(0)(2) always shifts negatively. This unusual behavior is discussed in terms of interplay between electrostatic interactions and conformational changes associated with cation binding. PMID- 12553840 TI - Ligand exchange dynamics in aluminum tris-(quinoline-8-olate): a solution state NMR study. AB - The exchange kinetics between the three symmetry-inequivalent ligands in the meridianal isomer of aluminum tris-(quinoline-8-olate) (Alq(3)), a widely used electron-transporting and light-emitting material in the field of organic light emitting diodes, have been studied using two-dimensional exchange NMR spectroscopy in solution. The three inequivalent ligands were found to exchange on a time scale of about 5 s(-)( 1) at room temperature. A simple first-order mechanism based on consecutive 180 degrees flips of the ligands is sufficient to quantitatively explain the experimental data. Activation enthalpies between 83 and 106 kJ mol(-)( 1) were found for the flips of the three inequivalent ligands. The activation entropies are positive, suggesting a highly disordered transition state. These findings elucidate the internally mobile nature of the Alq(3) complex, and may have important implications for the morphology of vapor deposited thin films of Alq(3) as well as for crystallization-assisted device failures. PMID- 12553841 TI - Chemical and photochemical electron transfer of new helianthrone derivatives: aspects of their photodynamic activity. AB - Helianthrones 2-4 are a new class of synthetic photosensitizers, which have a molecular skeleton related to that of hypericin. We established that irradiation of heliantrones with visible light leads to the formation of semiquinone radicals and reactive oxygen species. The structures of the paramagnetic anion species produced by electron transfer were calculated on the density functional level and investigated by cyclovoltammetry, UV/vis, and EPR/ENDOR spectroscopy. As with hypericin, the pi system of the helianthrones was found to be considerably deviated from planarity, and, upon electron transfer, deprotonation in the bay region occurs. The structure of the semiquinone radicals was found to be identical in THF, DMF, and aqueous buffered solutions regardless of the means by which reduction was achieved. Semiquinone radicals can be formed via self electron transfer between the excited state and the ground state or via electron transfer from an electron donor to the excited state of helianthrone. Therefore, the presence of an electron donor significantly enhanced the photogeneration of semiquinone and superoxide radical. The kinetic studies showed that no significant photochemical destruction of helianthrones occurred upon irradiation. Generation of superoxide and singlet oxygen upon irradiation of helianthrones was established by spin trapping techniques. This shows that both type I and type II mechanisms are of importance for the photodynamic action of these compounds. PMID- 12553842 TI - GFT NMR, a new approach to rapidly obtain precise high-dimensional NMR spectral information. AB - Widely used higher-dimensional Fourier transform (FT) NMR spectroscopy suffers from two major drawbacks: (i) The minimal measurement time of an N-dimensional FT NMR experiment, which is constrained by the need to sample N - 1 indirect dimensions, may exceed by far the measurement time required to achieve sufficient signal-to-noise ratios. (ii) The low resolution in the indirect dimensions severely limits the precision of the indirect chemical shift measurements. To relax on constraints arising from these drawbacks, we present here an acquisition scheme which is based on the phase-sensitive joint sampling of the indirect dimensions spanning a subspace of a conventional NMR experiment. This allows one to very rapidly obtain high-dimensional NMR spectral information. Because the phase-sensitive joint sampling yields subspectra containing "chemical shift multiplets", alternative data processing is required for editing the components of the multiplets. The subspectra are linearly combined using a so-called "G matrix" and subsequently Fourier-transformed. The chemical shifts are multiply encoded in the resonance lines constituting the shift multiplets. This corresponds to performing statistically independent multiple measurements, and the chemical shifts can thus be obtained with high precision. To indicate that a combined G-matrix and FT is employed, we named the new approach "GFT NMR spectroscopy". GFT NMR opens new avenues to establish high-throughput protein structure determination, to investigate systems with a higher degree of chemical shift degeneracy, and to study dynamic phenomena such as slow folding of biological macromolecules in greater detail. PMID- 12553843 TI - Structure of the aqueous solvated electron from resonance Raman spectroscopy: lessons from isotopic mixtures. AB - The structure and thermodynamics of the hydrated electron are probed with resonance Raman spectroscopy of isotopic mixtures of H(2)O and D(2)O. The strongly enhanced intramolecular bends of e(-)(H(2)O) and e(-)(D(2)O) produce single downshifted bands, whereas the e(-)(HOD) bend consists of two components: one slightly upshifted from the 1,446 cm(-1) bulk frequency to 1,457 cm(-1) and the other strongly downshifted to approximately 1,396 cm(-1). This 60 cm(-1) split and the 200 (120) cm(-1) downshifts of the OH (OD) stretch frequencies relative to bulk water reveal that the water molecules that are Franck-Condon coupled to the electron are in an asymmetric environment, with one proton forming a strong hydrogen bond to the electron. The downshifted bend and librational frequencies also indicate significantly weakened torsional restoring forces on the water molecules of e(-)(aq), which suggests that the outlying proton is a poor hydrogen bond donor to the surrounding solvent. A 1.6-fold thermodynamic preference of the electron for H(2)O is observed based on the relative intensities of the e(-)(H(2)O) and e(-)(D(2)O) bands in a 50:50 isotopic mixture. This equilibrium isotope effect is consistent with the downshifted vibrational frequencies and a relative reduction of the zero-point energy of H(2)O bound to the electron. Our results enhance the cavity model of the solvated electron and support only those models that contain water monomers as opposed to other molecular species. PMID- 12553844 TI - Normal and inverse primary kinetic deuterium isotope effects for C-H bond reductive elimination and oxidative addition reactions of molybdenocene and tungstenocene complexes: evidence for benzene sigma-complex intermediates. AB - The overall reductive elimination of RH from the ansa-molybdenocene and tungstenocene complexes [Me(2)Si(C(5)Me(4))(2)]Mo(Ph)H and [Me(2)Si(C(5)Me(4))(2)]W(R)H (R = Me, Ph) is characterized by an inverse primary kinetic isotope effect (KIE) for the tungsten system but a normal KIE for the molybdenum system. Oxidative addition of PhH to [[Me(2)Si(C(5)Me(4))(2)]M] also differs for the two systems, with the molybdenum system exhibiting a substantial intermolecular KIE, while no effect is observed for the tungsten system. These differences in KIEs indicate a significant difference in the reactivity of the hydrocarbon adducts [Me(2)Si(C(5)Me(4))(2)]M(RH) for the molybdenum and tungsten systems. Specifically, oxidative cleavage of [Me(2)Si(C(5)Me(4))(2)]M(RH) is favored over RH dissociation for the tungsten system, whereas RH dissociation is favored for the molybdenum system. A kinetics analysis of the interconversion of [Me(2)Si(C(5)Me(4))(2)]W(CH(3))D and [Me(2)Si(C(5)Me(4))(2)]W(CH(2)D)H, accompanied by elimination of methane, provides evidence that the reductive coupling step in this system is characterized by a normal KIE. This observation demonstrates that the inverse KIE for overall reductive elimination is a result of an inverse equilibrium isotope effect (EIE) and is not a result of an inverse KIE for a single step. A previous report of an inverse kinetic isotope effect of 0.76 for C-H reductive coupling in the [Tp]Pt(CH(3))H(2) system is shown to be erroneous. Finally, a computational study provides evidence that the reductive coupling of [Me(2)Si(C(5)Me(4))(2)]W(Ph)H proceeds via the initial formation of a benzene sigma-complex, rather than an eta(2)-pi-benzene complex. PMID- 12553845 TI - Protonation of gas-phase aromatic molecules: IR spectrum of the fluoronium isomer of protonated fluorobenzene. AB - The IR spectrum of the fluoronium isomer of protonated fluorobenzene (F C(6)H(6)F(+), phenylfluoronium) is recorded in the vicinity of the C-H and F-H stretch fundamentals to obtain the first structured spectrum of an isolated protonated aromatic molecule in the gas phase. Stable F-C(6)H(6)F(+) ions are produced via proton transfer from CH(5)(+) to fluorobenzene (C(6)H(5)F) in a supersonic plasma expansion. The F-C(6)H(6)F(+) spectrum recorded between 2,540 and 4,050 cm(-1) is consistent with a weakly bound ion-dipole complex composed of HF and the phenyl cation, HF-C(6)H(5)(+). The strongest transition occurs at 3,645 cm(-1) and is assigned to the F-H stretch (sigma(FH)). The antisymmetric C H stretch of the two ortho hydrogen atoms, sigma(CH) = 3,125 cm(-1), is nearly unshifted from bare C(6)H(5)(+), indicating that HF complexation has little influence on the C-H bond strength of C(6)H(5)(+). Despite the simultaneous production of the more stable ring protonated carbenium isomers of C(6)H(6)F(+) (fluorobenzenium) in the electron ionization source, F-C(6)H(6)F(+) can selectively be photodissociated into C(6)H(5)(+) and HF under the present experimental conditions, because it has a much lower dissociation energy than all carbenium isomers. Quantum chemical calculations at the B3LYP and MP2 levels of theory using the 6-311G(2df,2pd) basis support the interpretation of the experimental data and provide further details on structural, energetic, and vibrational properties of F-C(6)H(6)F(+), the carbenium isomers of C(6)H(6)F(+), and other weakly bound HF-C(6)H(5)(+) ion-dipole complexes. The dissociation energy of F-C(6)H(6)F(+) with respect to dehydrofluorination is calculated as D(0) = 4521 cm(-1) (approximately 54 kJ/mol). Analysis of the charge distribution in F-C(6)H(6)F(+) supports the notation of a HF-C(6)H(5)(+) ion-dipole complex, with nearly the whole positive charge of the added proton distributed over the C(6)H(5)(+) ring. As a result, protonation at the F atom strongly destabilizes the C-F bond in C(6)H(5)F. PMID- 12553848 TI - The rationale for using a topical retinoid for inflammatory acne. AB - Both comedogenesis and the development of inflammatory lesions in acne vulgaris appear to be related to genetic as well as immune processes. The key regulatory cytokine, interleukin-1alpha, has recently been documented as playing a major role in both the hypercornification and the orchestration of immune factors, ultimately resulting in noninflammatory and inflammatory lesions. Topical retinoids, such as tretinoin, and topical retinoid analogs, such as adapalene and tazarotene, help normalize the abnormal follicular keratinocyte desquamation - a key pathophysiologic factor in comedogenesis. This normalization also helps mitigate against the development of a propitious microenvironment for Propionibacterium acnes. Preclinical data suggest that topical retinoids and retinoid analogs may also have direct anti-inflammatory effects. A wealth of clinical data confirms that topical retinoids and retinoid analogs significantly reduce inflammatory lesions. Comparative clinical trials also demonstrate that adapalene has the best cutaneous tolerability profile of all these agents. Optimal therapy for inflammatory acne would involve the use of topical retinoids or retinoid analogs combined with oral or topical antibacterials. PMID- 12553846 TI - Toxicity screening by electrochemical detection of DNA damage by metabolites generated in situ in ultrathin DNA-enzyme films. AB - Rapid detection of DNA damage could serve as a basis for in vitro genotoxicity screening for new organic compounds. Ultrathin films (20-40 nm) containing myoglobin or cytochrome P450(cam) and DNA grown layer-by-layer on electrodes were activated by hydrogen peroxide, and the enzyme in the film generated metabolite styrene oxide from styrene. This styrene oxide reacted with double stranded (ds) DNA in the same film, mimicking metabolism and DNA damage in human liver. DNA damage was detected by square wave voltammetry (SWV) by using catalytic oxidation with Ru(bpy)(3)(2+) (bpy = 2,2'-bipyridine) and by monitoring the binding of Co(bpy)(3)(3+). Damaged DNA reacts more rapidly than intact ds-DNA with Ru(bpy)(3)(3+), giving SWV peaks at approximately 1 V versus SCE that grow larger with reaction time. Co(bpy)(3)(3+) binds more strongly to intact ds-DNA, and its SWV peaks at 0.04 V decreased as DNA was damaged. Little change in SWV signals was found for incubations of DNA/enzyme films with unreactive organic controls or hydrogen peroxide. Capillary electrophoresis and HPLC-MS suggested the formation of styrene oxide adducts of DNA bases under similar reaction conditions in thin films and in solution. The catalytic SWV method was more sensitive than the Co(bpy)(3)(3+) binding assay, providing multiple measurements over a 5 min reaction time. PMID- 12553850 TI - Darier's disease: epidemiology, pathophysiology, and management. AB - Darier's disease is a rare cutaneous disease with an autosomal dominant mode of inheritance. Greasy papules and plaques arise on the seborrheic areas and in the flexures and almost all patients have nail abnormalities. Acantholysis and dyskeratosis are the typical histological findings. The underlying defect is a result of mutations in the ATP2A2 gene on chromosome 12q23-24 that encodes for a sarco/endoplasmic reticulum calcium ATPase (SERCA 2). Acantholysis is thought to result from desmosome breakdown. Darier's disease is an example of a dominantly inherited disease caused by haplo-insufficiency. Oral retinoids are the most effective treatment but their adverse effects are troublesome. Topical retinoids, topical corticosteroids, surgery, and laser surgery have their advocates but evidence for efficacy is sparse. PMID- 12553849 TI - Ichthyosis: etiology, diagnosis, and management. AB - The ichthyoses are a heterogeneous group of disorders with both inherited and acquired forms. Clinical presentation, pattern of inheritance, and laboratory evaluation may establish a precise diagnosis, which can assist in prognosis and genetic counseling. Congenital autosomal recessive ichthyosis (CARI) usually presents at birth, often as a collodion baby. CARI can progress into any one of a spectrum of disorders. Lamellar ichthyosis is characterized by dark, plate (armor)-like scale. This disease is often caused by mutations in the gene encoding the enzyme transglutaminase 1. Congenital ichthyosiform erythroderma is another phenotype within CARI, marked by generalized redness and fine white scale. Epidermolytic hyperkeratosis is an autosomal dominant disorder characterized by hyperkeratosis and blistering, and at least six clinical phenotypes have been described. It may be due to mutations in the gene encoding the intermediate filament proteins keratin 1 and 10. Ichthyosis vulgaris is the most common ichthyosis, and is inherited in an autosomal dominant pattern. Involvement is generally mild and may vary greatly with climate and humidity. X linked ichthyosis, due to a defect in the enzyme steroid sulfatase, affects males with generalized scaling that usually begins soon after birth. There may be associated corneal opacities that do not affect vision. Sjogren-Larsson syndrome is an autosomal recessive ichthyosis associated with progressive spastic paralysis and mental retardation. This condition is caused by mutations in the gene for fatty aldehyde dehydrogenase. Refsum's disease, due to accumulation of phytanic acid, results in ichthyosis and progressive neurologic dysfunction. The erythrokeratodermas are characterized by hyperkeratosis and localized erythema. Erythrokeratodermia variabilis is autosomal dominant and characterized by generalized or localized hyperkeratosis and migratory red patches. Mutations in the genes encoding the gap junction proteins, connexins, underlie this disorder. Netherton's syndrome is an autosomal recessive disorder characterized by ichthyosis, a hair shaft abnormality and atopy. The ichthyosis may present at birth with erythroderma or in some cases a collodion presentation. However, a frequent characteristic skin manifestation is ichthyosis linearis circumflexa. Netherton's syndrome has been found to be due to an abnormality in a serum protease inhibitor. Acquired ichthyosis can have a variety of underlying causes including neoplastic, infectious, drugs, endocrine, metabolic, autoimmune, malabsorptive states, and hereditary. Topical, and in more severe cases, systemic, therapy are useful in managing this array of disorders of cornification. PMID- 12553851 TI - Ceramides and skin function. AB - Ceramides are the major lipid constituent of lamellar sheets present in the intercellular spaces of the stratum corneum. These lamellar sheets are thought to provide the barrier property of the epidermis. It is generally accepted that the intercellular lipid domain is composed of approximately equimolar concentrations of free fatty acids, cholesterol, and ceramides. Ceramides are a structurally heterogeneous and complex group of sphingolipids containing derivatives of sphingosine bases in amide linkage with a variety of fatty acids. Differences in chain length, type and extent of hydroxylation, saturation etc. are responsible for the heterogeneity of the epidermal sphingolipids. It is well known that ceramides play an essential role in structuring and maintaining the water permeability barrier function of the skin. In conjunction with the other stratum corneum lipids, they form ordered structures. An essential factor is the physical state of the lipid chains in the nonpolar regions of the bilayers. The stratum corneum intercellular lipid lamellae, the aliphatic chains in the ceramides and the fatty acids are mostly straight long-chain saturated compounds with a high melting point and a small polar head group. This means that at physiological temperatures, the lipid chains are mostly in a solid crystalline or gel state, which exhibits low lateral diffusional properties and is less permeable than the state of liquid crystalline membranes, which are present at higher temperatures. The link between skin disorders and changes in barrier lipid composition, especially in ceramides, is difficult to prove because of the many variables involved. However, most skin disorders that have a diminished barrier function present a decrease in total ceramide content with some differences in the ceramide pattern. Formulations containing lipids identical to those in skin and, in particular, some ceramide supplementation could improve disturbed skin conditions. Incomplete lipid mixtures yield abnormal lamellar body contents, and disorder intercellular lamellae, whereas complete lipid mixtures result in normal lamellar bodies and intercellular bilayers. The utilization of physiological lipids according to these parameters have potential as new forms of topical therapy for dermatoses. An alternative strategy to improving barrier function by topical application of the various mature lipid species is to enhance the natural lipid-synthetic capability of the epidermis through the topical delivery of lipid precursors. PMID- 12553852 TI - Effects of alefacept on health-related quality of life in patients with psoriasis: results from a randomized, placebo-controlled phase II trial. AB - INTRODUCTION: Chronic plaque psoriasis has a profound impact on patient quality of life (QOL), including adverse psychosocial effects, impaired daily activities, anxiety, and depression. OBJECTIVE: To assess health-related QOL in a randomized phase II trial of alefacept (human LFA-3/IgG(1) fusion protein), a selective immunomodulator for psoriasis. STUDY DESIGN: Multicenter, randomized, placebo controlled, double-blind trial. METHODS: 229 patients with moderate to severe psoriasis were randomized to alefacept (0.025, 0.075, or 0.150 mg/kg) or placebo by 30-second intravenous bolus once weekly for 12 weeks and followed for 12 additional weeks. Patients completed a general (SF-36) Health Survey) and dermatology-specific (Dermatology Life Quality Index [DLQI] and Dermatology Quality Of Life Scales [DQOLS]) QOL surveys at each study visit. RESULTS: Patients treated with alefacept had significantly greater improvements on dermatology-specific QOL scales compared with patients receiving placebo (p < 0.05). Patients who achieved a >or=50% or a >or=75% reduction in Psoriasis Area and Severity Index (PASI) reported similar improvement in QOL, which was significantly greater than that of other patients. CONCLUSIONS: The clinical effect of alefacept on psoriasis is associated with an improvement in patients' QOL. Among patients with moderate to severe psoriasis, an improvement in PASI of 50% or more is associated with better QOL scores. PMID- 12553854 TI - Characterization of rhodamine conjugated Agiotensin II peptide: synthesis, analysis and receptor binding and internalization. AB - The results in this study show that the rhodamine fluorophore can be specifically conjugated to Angiotensin II at Lys3 residue (substituted for a Val) without altering the biological activity of the parent compound. The conjugated peptide was characterized using HPLC, mass spectrometry, and N-terminal sequencing. The rhodamine-Angiotensin II binds effectively to AT1 receptor and gets internalized in clathrin coated vesicles by endocytosis. These results clearly suggest the usefulness of fluorophore-conjugated peptides in studies such as, ligand-receptor binding, and ligand-receptor complex internalization, for drug delivery using cell receptors and as an alternative to peptide hormone radioimmunoassays. PMID- 12553855 TI - Virus-cell fusion inhibitory activity of novel analogue peptides based on the HP (2-20) derived from N-terminus of Helicobacter pylori Ribosomal Protein L1. AB - HP (2-20) (AKKVFKRLEKLFSKIQNDK) is the antibacterial sequence derived from N terminus of Helicobacter pylori Ribosomal Protein L1 (RPL1). It has a broad spectrum microbicidal activity in vitro that is thought to be related to the membrane-disruptive properties of the peptide. Based on the putative membrane targeted mode of action, we postulated that HP (2-20) might be possessed virus cell fusion inhibitory activity. To develop the novel virus-cell fusion inhibitory peptides, several analogues with amino acid substitution were designed to increase or decrease only net hydrophobic region. In particular, substitution of Gln and Asp for hydrophobic amino acid, Trp at position 17 and 19 of HP (2-20) (Anal 3) caused a dramatic increase in virus-cell fusion inhibitory activity without hemolytic effect. PMID- 12553856 TI - Structure-antibacterial activity relationship of cecropin A derivatives. AB - To investigate the effect of net positive charge, alpha-helicity and hydrophobicity of the peptides on antibacterial activity, we designed three kinds of cecropin A (CA) derivatives. Compared to CA, F3 with the highest net positive charge exhibited similar or slightly weaker antibacterial activity (MIC: 3.13 approximately 6.25 microM). F1 showed lower antibacterial activity than that of F3, even though it has higher hydrophobicity and a-helicity than F3. This result indicates that the net positive charge of cecropin A-like peptides appears to be a more important factor in antibacterial activity than alpha-helicity and hydrophobicity. The two peptides F1 and F2 possessed almost similar antibacterial activity, but F2 showed very lower activity in the membrane disruption than F1, suggesting other factors are involved in the antibacterial activity of the peptides as well as peptide-lipid interaction. PMID- 12553857 TI - A model for the cytoplasmic domain of the influenza A virus M2 channel by analogy to the HIV-1 Vpu protein. AB - A model for the cytoplasmic domain of the M2 proton channel of influenza A virus was formulated based primarily on the cytoplasmic domain of the Vpu protein of HIV-1 using sequence similarity and structure prediction techniques. The model consists of a pair of antiparallel helices followed by a strand parallel to the first helix. Structural analogies with other proteins contribute support for features of the model and suggest ways in which the M2 cytoplasmic domain can interact with other viral and cellular factors. PMID- 12553858 TI - Selective precipitation of haptoglobin and alpha2-macroglobulin from human serum using Alocasia macrorhiza tuber protein. AB - Treatment of human serum with ammonium sulfate fraction (0-50%) of Alocasia macrorhiza tuber extract resulted in precipitation at neutral pH. The precipitate was dissolved at pH 10.5 and chromatographed on Sephadex G-100 column. Two protein peaks were resolved. While the first peak represented alpha2 macroglobulin and haptoglobin, the second peak accounted for specific Alocasia protein. Incidentally the Alocasia protein was shown to be responsible for selective and specific precipitation of alpha2-macroglobulin and haptoglobin from serum. Thus the plant protein in its pure form or in crude stage could be used for the rapid isolation of two of the prominent alpha2-globulins. PMID- 12553859 TI - Biochemical characterization of apoptotic cleavage of KH-type splicing regulatory protein (KSRP)/far upstream element-binding protein 2 (FBP2). AB - Caspases, Asp-specific cysteine protease, cleave proteins upon apoptosis. To identify and characterize new caspase substrate in the nucleus, the proteome of the rat liver extracts was analyzed after the treatment with caspases. One of the identified proteins was KSRP/FBP2 that is preferentially cleaved by caspase-3 and 7 at two sites after Asp102 and Asp183. The second site was cleaved only in the protein produced in cells, but not in in vitro translated protein. These results indicate that more than the primary sequence may be important for the recognition by caspases. PMID- 12553860 TI - Acyl-gelatins for cell-hybrid biomaterials: preparation of gelatins with high melting point and affinity for hydrophobic surfaces. AB - In the development of cell-hybrid biomaterials, the functional activity of cells depends on the selective binding of cells to artificial ligands on the biomaterials. The extracellular matrix (ECM) is the most important ligand for cell activity. ECM is known to contain collagen, one of whose constituents is gelatin. Although natural gelatin has good cell attachment properties, the melting point of gelatin hydrogel is lower than body temperature. Thus, non chemically cross-linked gelatin hydrogel is not a biomaterial that is used for prostheses. In the present study, we report the preparation of acyl-gelatin hydrogels with high melting point (>37 degrees C) and high affinity for hydrophobic surfaces for easy handling for transportation and adhesion activities on the hydrophobic surfaces. In addition, the doubling time of endothelial cells on the coated cell culture plate was faster than that of natural gelatin owing to the higher adhesion activity of acyl-gelatin. The results clearly demonstrated that the acyl-gelatin acted as an interface that enabled cell adhesion to artificial materials surfaces. PMID- 12553861 TI - Synthesis of alkyl and aryl esters of N-protected beta-homoamino acids from N protected alpha-aminodiazoketones. AB - A simple and concomitant esterification method for the synthesis of methyl, ethyl, t-butyl, benzyl, and 9-fluorenylmethyl esters of Fmoc-/Boc-/Z-beta homoamino acids employing Fmoc-/Boc-/Z-alpha-aminodiazoketones by Wolff rearrangement is described. The method offers good yield with purity. PMID- 12553862 TI - Antibiotic properties of novel synthetic temporin A analogs and a cecropin A temporin A hybrid peptide. AB - Temporin A, 18 analogs, and a cecropin A-temporin A hybrid peptide were tested with antibiotic sensitive and resistant bacteria, fungi, human erythrocytes, and in clotting assays. Several peptides were active in these assays, and some analogs (D-TA, W1-TA, and Con-L4,G10) may be useful lead compounds for further antibiotics development. The activity of temporin A was found to be dependent upon several of its structural features, including amino acid composition and sequence, chirality, helicity, and positive charge. PMID- 12553863 TI - A direct continuous pH-spectrophotometric assay for arginine kinase activity. AB - A direct and continuous spectrophotometric method was developed for determining arginine kinase (phosphoarginine synthesis) activity. Protons are produced during the phosphoarginine synthesis course, so adding the complex acid-base indicator to this solution and monitoring the decrease of absorbance of the solution at 575 nm will follow the arginine kinase activity. For this condition, one activity unit of arginine kinase was defined as 1 micromol H+ produced in 1 minute in the enzymatic reaction catalyzed by arginine kinase. PMID- 12553864 TI - Purification, crystallization and preliminary X-ray studies of human augmenter of liver regeneration. AB - Human augmenter of liver regeneration has been expressed in Escherichia coli, purified and crystallized. The crystals belong to space group C222, with unit cell parameters a=51.7 A, b=78.8 A, c=63.7 A. Diffraction data were collected to 2.80 A with a completeness of 99.9% (99.9% for the last shell), a R(sym) value of 0.092(0.236) and an I/sigma(I) value of 6.2(2.7). PMID- 12553865 TI - Fusion pore or porosome: structure and dynamics. AB - Electrophysiological measurements on live secretory cells almost a decade ago suggested the presence of fusion pores at the cell plasma membrane. Membrane bound secretory vesicles were hypothesized to dock and fuse at these sites, to release their contents. Our studies using atomic force microscopy on live exocrine and neuroendocrine cells demonstrate the presence of such plasma membrane pores, revealing their morphology and dynamics at near nm resolution and in real time. PMID- 12553866 TI - Effects of cortisol and oestradiol on hepatic 11beta-hydroxysteroid dehydrogenase type 1 and glucocorticoid receptor proteins in late-gestation sheep fetus. AB - In the late-gestation sheep, increased fetal plasma cortisol concentration and placental oestradiol (E(2)) output contribute to fetal organ maturation, in addition to the onset of parturition. Both cortisol and E(2) are believed to regulate the enzyme 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1), which interconverts bioactive 11-hydroxy glucocorticoids and their inactive 11 keto metabolites. 11beta-HSD1, abundantly expressed in fetal liver, operates primarily as a reductase enzyme to produce bioactive cortisol and thus regulates local hepatic glucocorticoid concentrations. Cortisol acts through the glucocorticoid receptor (GR) present in the liver. In this study, we examined the effects of cortisol and E(2) on hepatic 11beta-HSD1 and GR in the liver of chronically catheterized sheep fetuses treated with saline (n=5), cortisol (1.35 mg/h; n=5), saline+4-hydroxyandrostendione, a P450 aromatase inhibitor (4-OHA; 1.44 mg/h; n=5), or cortisol+4-OHA (n=5). Cortisol infusion resulted in increased plasma concentrations of fetal cortisol and E(2); concurrent administration of 4 OHA attenuated the increase in plasma E(2) concentrations. Using immunohistochemistry, we showed that fetal hepatocytes expressed both 11beta-HSD1 and GR proteins. Cortisol treatment increased GR in both cytosol and nuclei of hepatocytes; concurrent administration of 4-OHA was associated with distinct nuclear GR staining. Western blot revealed that cortisol, in the absence of increased E(2) concentrations, significantly increased concentrations of 11beta HSD1 (34 kDa) and GR (95 kDa) proteins. 11beta-HSD1 enzyme activity was measured in the liver microsomal fraction in the presence of [(3)H]cortisone (10(-)(6) M) or [(3)H]cortisol (10(-)(6) M) and NADPH (reductase activity) or NADP(+) (dehydrogenase activity) respectively. 11beta-HSD1 reductase activity was significantly greater in the presence of cortisol. In summary, we found that, in sheep during late gestation, cortisol increased both 11beta-HSD1 and GR in the fetal liver, and these effects were accentuated in the absence of increased E(2). PMID- 12553867 TI - Enhanced 11beta-hydroxysteroid dehydrogenase type 1 activity in stress adaptation in the guinea pig. AB - The 11beta-hydroxysteroid dehydrogenases (11beta-HSDs) convert cortisol to its inactive metabolite cortisone and vice versa. 11beta-HSD type 1 (11beta-HSD-1) functions as a reductase in vivo, regulating intracellular cortisol levels and its access to the glucocorticoid receptor. In contrast, 11beta-HSD-2 only mediates oxidation of natural glucocorticoids, and protects the mineralocorticoid receptor from high cortisol concentrations. We investigated the in vivo and in vitro effects of ACTH on the recently characterized 11beta-HSDs in guinea pig liver and kidney. Tissue slices of untreated guinea pigs were incubated with (3)H labelled cortisol or cortisone and ACTH(1-24) (10(-10) and 10(-9) mol/l). The 11beta-HSD activities in liver and kidney slices were not influenced by in vitro incubation with ACTH(1-24). In addition, guinea pigs were treated with ACTH(1-24) or saline injections s.c. for 3 days. Liver and kidney tissue slices of these animals were incubated with (3)H-labelled cortisol or cortisone. In vivo ACTH treatment significantly increased reductase and decreased oxidase activity in liver and kidney. Furthermore, 11beta-HSD-1 activity assessed by measurement of the urinary ratio of (tetrahydrocortisol (THF)+5alphaTHF)/(tetrahydrocortisone) was significantly increased after ACTH treatment compared with the control group. Plasma levels of cortisol, cortisone, progesterone, 17-hydroxyprogesterone and androstenedione increased significantly following in vivo ACTH treatment. The enhanced reductase activity of the hepatic and renal 11beta-HSD-1 is apparently caused by cortisol or other ACTH-dependent steroids rather than by ACTH itself. This may be an important fine regulation of the glucocorticoid tonus for stress adaptation in every organ, e.g. enhanced gluconeogenesis in liver. PMID- 12553868 TI - Regulation of serum leptin and its role in the hyperphagia of lactation in the rat. AB - The factors regulating serum leptin concentration and its relationship to the hyperphagia of lactation have been investigated in rats. Lactation results in hypoleptinaemia and loss, or at least marked attenuation, of the nocturnal rise in serum leptin. Litter removal resulted in a fall in food intake and restoration of the nocturnal rise in serum leptin. Returning the litter to the mother after a 48-h absence increased food intake and began to reinitiate milk production, but the nocturnal serum leptin levels were still increased at 48 h after litter restoration. Adjusting litter size to four, eight, ten or fourteen pups at parturition resulted in different rates of litter growth and food intake during the subsequent lactation, but had no effect on the degree of hypoleptinaemia. Reducing litter size from ten to four pups at mid-lactation resulted in a transient increase in both serum leptin and pup growth rate, while food intake fell to a level found in rats suckling four pups throughout lactation. Reducing milk production by injection of bromocriptine increased serum leptin, but did not restore the nocturnal rise in serum leptin; food intake decreased, but remained much higher than in non-lactating rats. Feeding a varied, high-energy diet resulted in a decrease in the weight of food ingested, but no change in calorie intake, and had no effect on the hypoleptinaemia. These studies suggested that the hypoleptinaemia of lactating rats is due to negative energy balance, but the loss of the nocturnal rise in serum leptin is due to the suckling stimulus. The negative energy balance of lactation does not appear to be caused by a physical constraint on food intake. While the hypoleptinaemia should facilitate the hyperphagia of lactation, other orexigenic signals must also be involved. PMID- 12553869 TI - Insulin restores GH responsiveness during lactation-induced negative energy balance in dairy cattle: effects on expression of IGF-I and GH receptor 1A. AB - Early lactation in dairy cattle is a period of severe negative energy balance (NEB) characterized by reduced blood glucose and insulin concentrations and elevated blood GH concentrations. The liver is refractory to GH during NEB and this uncoupling of the GH-IGF axis results in diminished plasma concentrations of IGF-I. Our objectives were to examine the effects of insulin administration during the immediate postpartum period on plasma IGF-I and GH concentrations and to examine the hepatic expression of total GH receptors (all GH receptor transcripts), GH receptor 1A (GHR 1A) and IGF-I. In addition, we examined adipose tissue for total GH receptor and IGF-I mRNA levels to establish the effects of chronic hyperinsulinemia on an insulin-responsive peripheral tissue. Holstein cows (n=14) were subjected to either a hyperinsulinemic-euglycemic clamp (insulin; INS) or saline infusion (control; CTL) for 96 h starting on day 10 postpartum. Insulin was infused i.v. (1 micro g/kg body weight per h), blood samples were collected hourly, and euglycemia was maintained by infusion of glucose. Insulin concentrations during the infusions were increased 8-fold in INS compared with CTL cows (2.33+/-0.14 vs 0.27+/-0.14 ng/ml (S.E.M.); P<0.001) while blood glucose concentrations were not different between treatments (45.3+/-2.2 vs 42.5+/-2.2 mg/dl; P>0.1). Plasma IGF-I increased continuously during the insulin infusion, and reached the highest concentrations at the end of the clamp, being almost 4-fold higher in INS compared with CTL cows (117+/-4 vs 30+/-4 ng/ml; P<0.001). Hepatic expression of GHR 1A and IGF-I mRNA was low in CTL cows, but was increased 3.6-fold (P<0.05) and 6.3-fold (P<0.001) respectively in INS cows. By contrast, in adipose tissue the changes in gene expression in response to insulin were reversed with decreases in both total GHR and IGF-I mRNA. The expressions of GHR 1A and IGF-I mRNA in liver tissue were correlated in INS (r=0.86; P<0.05), but not CTL cows (r=0.43; P>0.1). Insulin appears to be a key metabolic signal in coupling the GH-IGF axis, thus orchestrating a marked elevation in circulating IGF-I concentrations. PMID- 12553870 TI - The GH/IGF-I axis in the brushtail possum (Trichosurus vulpecula) pouch young. AB - Plasma and pituitary GH concentrations and liver GH receptor (GHR), IGF-I and IGF binding protein-3 (IGFBP-3) mRNA expression were determined in brushtail possum (Trichosurus vulpecula) pouch young aged 12-150 days post-partum and in adults. Mean plasma GH concentrations were highest, measuring around 150 ng/ml, from 12 to 100 days post-partum, and thereafter declined so that by 150 days post-partum levels were not significantly different from those in adults (10.8+/-1.8 ng/ml (S.E.M.)). In contrast to plasma levels, pituitary GH content increased markedly throughout pouch life, with an 87-fold increase between 12 and 150 days post partum. However, when expressed per gram body weight, pituitary content was relatively constant between 25 and 150 days post-partum, indicating that the decline in plasma GH after 100 days post-partum was not due to decreased synthesis and/or storage of GH in the pituitary gland. Expression of GHR, IGF-I and IGFBP-3 mRNAs was determined by semi-quantitative RT-PCR. Liver GHR and IGF-I mRNA expression were low at 12 and 25 days post-partum and did not show sustained and significant increases (P<0.05) until 125 and 150 days post-partum. IGFBP-3 expression was also low at 12 days post-partum but then increased rapidly to a maximum at 50 days post-partum and thereafter declined. For all three mRNAs, liver expression at day 150 was not significantly different from that in adults. These patterns of gene expression for GHR and IGF-I suggest that the possum liver is resistant to the high plasma GH concentrations during early pouch life and in this way is similar to the fetal liver of some eutherian mammals. PMID- 12553871 TI - Effect of recombinant porcine IGF-binding protein-3 on proliferation of embryonic porcine myogenic cell cultures in the presence and absence of IGF-I. AB - IGF-binding protein (IGFBP)-3 is produced by cultured porcine embryonic myogenic cell (PEMC) cultures and is secreted into the medium. Levels of secreted IGFBP-3 and IGFBP-3 mRNA are significantly reduced during differentiation and increase after differentiation is complete, suggesting that IGFBP-3 may play some role in myogenesis and/or in changes in myogenic cell proliferation that accompany differentiation. IGFBP-3 reportedly may either suppress or stimulate proliferation of cultured cells depending on cell type. Additionally, IGFBP-3 has been shown to affect proliferation via both IGF-dependent and IGF-independent mechanisms in some cell types but not all. Currently, the effect, if any, of IGFBP-3 on myogenic cell proliferation is not known. Consequently, the goal of this study was to assess the IGF-I-dependent and IGF-I-independent actions of recombinant porcine IGFBP-3 on proliferation of cultured porcine myogenic cells. To facilitate these investigations, we have expressed porcine IGFBP-3 in the baculovirus system, purified and characterized the expressed recombinant porcine IGFBP-3 (rpIGFBP-3), and produced and characterized an anti-porcine IGFBP-3 antibody that neutralizes the biological activity of porcine IGFBP-3. rpIGFBP-3 suppressed IGF-I-stimulated proliferation of PEMCs in a concentration-dependent manner with equimolar concentrations of IGF-I and rpIGFBP-3, resulting in complete suppression of IGF-I-stimulated proliferation. rpIGFBP-3 also suppressed Long-R3-IGF-I-stimulated proliferation of PEMC, indicating that rpIGFBP-3 possesses IGF-independent activity in this cell system. These data have established that IGFBP-3 has the potential to affect proliferation of PEMCs during critical periods of muscle development that may impact ultimate muscle mass achievable postnatally. PMID- 12553872 TI - Progressive development of insulin resistance phenotype in male mice with complete aromatase (CYP19) deficiency. AB - Aromatase (CYP19) is a cytochrome P450 enzyme that catalyzes the formation of aromatic C18 estrogens from C19 androgens. It is expressed in various tissues and contributes to sex-specific differences in cellular metabolism. We have generated aromatase-knockout (ArKO) mice in order to study the role of estrogen in the regulation of glucose metabolism. The mean body weights of male ArKO (-/-) mice (n=7) and wild-type littermates (+/+) (n=7) at 10 and 12 weeks of age were 26.7+/ 1.9 g vs 26.1+/-0.8 g and 28.8+/-1.4 g vs 26.9+/-1.0 g respectively. The body weights of the ArKO and wild-type mice diverged between 10 and 12 weeks of age with the ArKO males weighing significantly more than their wild-type littermates (P<0.05). The ArKO males showed significantly higher blood glucose levels during an intraperitoneal glucose tolerance test compared with wild-type littermates beginning at 18 weeks of age. By 24 weeks of age, they had higher fasting blood glucose levels compared with wild-type littermates (133.8+/-22.8 mg/dl vs 87.8+/ 20.3 mg/dl respectively; P<0.01). An intraperitoneal injection of insulin (0.75 mU insulin/g) caused a continuous decline in blood glucose levels in wild-type mice whereas ArKO males at 18 weeks and older exhibited a rebound increase in glucose levels 30 min after insulin injection. Thus, ArKO male mice appear to develop glucose intolerance and insulin resistance in an age-dependent manner. There was no difference in fasting serum triglyceride and total cholesterol levels between ArKO male mice and wild-type littermates at 13 and 25 weeks of age. However, serum triglyceride and cholesterol levels were significantly elevated following a meal in ArKO mice at 36 weeks of age. Serum testosterone levels in ArKO male mice were continuously higher compared with wild-type littermates. Treatment of ArKO males with 17beta-estradiol improved the glucose response as measured by intraperitoneal glucose and insulin tolerance tests. Treatment with fibrates and thiazolidinediones also led to an improvement in insulin resistance and reduced androgen levels. As complete aromatase deficiency in man is associated with insulin resistance, obesity and hyperlipidemia, the ArKO mouse may be a useful animal model for examining the role of estrogens in the control of glucose and lipid homeostasis. PMID- 12553873 TI - Maternal hypothyroidism in the rat influences placental and liver glycogen stores: fetal growth retardation near term is unrelated to maternal and placental glucose metabolic compromise. AB - Maternal hypothyroidism impairs fetal growth in the rat, but the mechanisms by which this occurs are unknown. Since the fetus derives its glucose supply from the mother, and maternal thyroidectomy may disturb maternal and placental glucose metabolism, we postulated that maternal and/or placental glucose metabolic compromise may contribute to fetal growth retardation in hypothyroid dams. Feto placental growth, tissue glycogen stores and glucose levels in sera and amniotic fluid were determined in rat dams partially thyroidectomized (TX) before pregnancy and in euthyroid controls. Fetal body weight at 16, 19 and 21 days gestation (d.g.) was related to pre-mating maternal serum total thyroxine (TT(4)) levels; permanent fetal growth retardation occurred in severely (TX(s); pre mating maternal serum TT(4) GnRH>>chicken (c) GnRH-II), and mediated stimulation of [(3)H]inositol phosphate (IP) accumulation. Increasing viral titre from 3 to 300 p.f.u./cell increased receptor number (2000 to 275 000 sites/cell respectively) and [(3)H]IP responses. GnRH also caused a biphasic increase in the cytoplasmic Ca(2+) concentration in Ad GnRH-R-infected cells but not in control cells. Mobilization of Ca(2+) from intracellular stores contributed to the spike phase of this response whereas the plateau phase was dependent upon Ca(2+) entry across the plasma membrane. This effect on Ca(2+) and stimulation of [(3)H]IP accumulation were both blocked by the GnRH-R antagonist, Cetrorelix. In addition, GnRH reduced cell number (as measured in MTT activity assays) and DNA synthesis (as measured by [(3)H]thymidine incorporation) in Ad GnRH-R-infected cells (but not in control cells). This effect was mimicked by agonist analogues and inhibited by two antagonists. Thus, when exogenous GnRH-Rs are expressed at a density comparable to that in gonadotrophs, they are functionally indistinguishable from the endogenous GnRH-Rs in gonadotrophs. Moreover, expression of high affinity GnRH-Rs can facilitate a direct anti-proliferative effect of GnRH agonists on prostate cancer cells. PMID- 12553877 TI - Expression and role of mannose receptor/terminal high-mannose type oligosaccharide on osteoclast precursors during osteoclast formation. AB - Osteoclasts are formed from hematopoietic precursors via cell-cell fusion. We have previously reported that mannose residues are expressed on the outer membranes of monocytes during osteoclast differentiation. In the present study, we have attempted to demonstrate the pattern of expression levels of terminal high-mannose type oligosaccharide and to show that the mannose receptor is expressed on osteoclast precursor cells. Osteoclasts were formed using three different systems, namely mouse bone marrow cell culture, co-culture of mouse spleen cells with stromal cells, and RAW264.7 cell cultures. During osteoclast differentiation, the expression of terminal high-mannose type oligosaccharide gradually increased and then peaked at the stage of fusion in all three systems. Expression of the mannose receptor gradually increased during osteoclast differentiation in bone marrow cells and the co-culture system. In contrast, that in RAW264.7 cells had already been detected in the absence of the soluble receptor activator of NF-kappaB ligand and did not change during osteoclast differentiation. To ascertain whether expression of high-mannose type oligosaccharide is involved in tartrate-resistant acid phosphatase (TRAP) positive multinucleated cell (MNC) formation, glycosidase inhibitors were used on RAW264.7 cell culture. Castanospermine, an inhibitor of glucosidase I, inhibited the TRAP-positive MNCs, and deoxymannojirimycin, an inhibitor of alpha mannosidase I, increased the TRAP-positive MNC formation. These results indicate that the binding of terminal high-mannose and mannose receptor is important for the process of cellular fusion in osteoclast formation. PMID- 12553878 TI - The heparin-binding 10 kDa fragment of connective tissue growth factor (CTGF) containing module 4 alone stimulates cell adhesion. AB - Connective tissue growth factor (CTGF) is a 349-residue mosaic protein that contains four structural modules implicated in protein-protein interactions. To address the functionality of residues 247-349 (containing module 4 alone), this region of CTGF was produced as a maltose binding protein (MBP) fusion protein in E. coli. After removal of MBP, recombinant CTGF commenced at Glu(247), was of M(r) 10 000, was immunoreactive with anti-CTGF[247-260], bound strongly to heparin, and promoted dose-dependent adhesion of fibroblasts, myofibroblasts, endothelial cells, and epithelial cells. An 8 kDa presumptive C-terminally truncated form of CTGF commencing at Glu(247) also promoted cell adhesion. CTGF mediated cell adhesion was abolished by heparin or EDTA. These data demonstrate the presence of heparin-binding and cell-adhesion motifs within the C-terminal 103 residues of CTGF and show that CTGF-mediated cell adhesion is heparin-and divalent cation-dependent. Thus, CTGF isoforms comprising essentially module 4 are intrinsically functional in the absence of the other constituent modules of CTGF. PMID- 12553879 TI - Iron-sulphur cluster assembly in plants: distinct NFU proteins in mitochondria and plastids from Arabidopsis thaliana. AB - Recent results are in favour of a role for NFU-like proteins in Fe-S cluster biogenesis. These polypeptides share a conserved CXXC motif in their NFU domain. In the present study, we have characterized Arabidopsis thaliana NFU1-5 genes. AtNFU proteins are separated into two classes. NFU4 and NFU5 are part of the mitochondrial type, presenting a structural organization similar to Saccharomyces cerevisiae Nfu1p. These proteins complement a Delta isu1 Delta nfu1 yeast mutant and NFU4 mitochondrial localization was confirmed by green fluorescent protein fusion analysis. AtNFU1-3 represent a new class of NFU proteins, unique to plants. These polypeptides are made of two NFU domains, the second having lost its CXXC motif. AtNFU1-3 proteins are more related to Synechocystis PCC6803 NFU like proteins and are localized to plastids when fused with the green fluorescent protein. NFU2 and/or NFU3 were detected in leaf chloroplasts by immunoblotting. NFU1 and NFU2 are functional NFU capable of restoring the growth of a Delta isu1 Delta nfu1 yeast mutant, when addressed to yeast mitochondria. Furthermore, NFU2 recombinant protein is capable of binding a labile 2Fe-2S cluster in vitro. These results demonstrate the presence of distinct NFU proteins in Arabidopsis mitochondria and plastids. Such results suggest the existence of two different Fe S assembly machineries in plant cells. PMID- 12553880 TI - Attenuation of glucocorticoid functions in an Anx-A1-/- cell line. AB - The Ca(2+)- and phospholipid-binding protein Anx-A1 (annexin 1; lipocortin 1) has been described both as an inhibitor of phospholipase A(2) (PLA(2)) activity and as a mediator of glucocorticoid-regulated cell growth and eicosanoid generation. Here we show that, when compared with Anx-A1(+/+) cells, lung fibroblast cell lines derived from the Anx-A1(-/-) mouse exhibit an altered morphology characterized by a spindle-shaped appearance and an accumulation of intracellular organelles. Unlike their wild-type counterparts, Anx-A1(-/-) cells also overexpress cyclo-oxygenase 2 (COX 2), cytosolic PLA(2) and secretory PLA(2) and in response to fetal calf serum, exhibit an exaggerated release of eicosanoids, which is insensitive to dexamethasone (10(-8)- 10(-6) M) inhibition. Proliferation and serum-induced progression of Anx-A1(+/+) cells from G(0)/G(1) into S phase, and the associated expression of extracellular signal-regulated kinase 2 (ERK2), cyclin-dependent kinase 4 (cdk4) and COX 2, is strongly inhibited by dexamethasone, whereas Anx-A1(-/-) cells are refractory to the drug. Loss of the response to dexamethasone in Anx-A1(-/-) cells occurs against a background of no apparent change in glucocorticoid receptor expression or sensitivity to non-steroidal anti-inflammatory drugs. Taken together, these observations suggest strongly that Anx-A1 functions as an inhibitor of signal transduction pathways that lead to cell proliferation and may help to explain how glucocorticoids regulate these processes. PMID- 12553881 TI - Fatty acids liberated from high-density lipoprotein phospholipids by endothelial derived lipase are incorporated into lipids in HepG2 cells. AB - We previously reported that endothelial-derived lipase (EDL) efficiently hydrolyses high-density-lipoprotein-derived phosphatidycholine (HDL-PC). In the present study, we assessed the ability of EDL to supply HepG2 cells with non esterified fatty acids (NEFA) liberated from HDL-phospholipids. For this purpose, HepG2 cells infected with adenovirus encoding human EDL (EDL-Ad), or with control beta-galactosidase-expressing adenovirus (LacZ-Ad), were incubated with (14)C-HDL PC. The analysis of the cellular lipids by TLC revealed that EDL overexpression led to an increase in the amount of cellular (14)C-lipids, whereby the label was mainly incorporated into phospholipids and triacylglycerols (TAG). Cells expressing mutant enzymically inactive EDL (MUT-EDL-Ad) contained similar amounts of (14)C-TAG but higher amounts of (14)C-phosphatidylcholine (PC) compared with LacZ-Ad-infected cells. The co-expression of CD36 augmented the EDL-mediated accumulation of (14)C-lipids in HEK-293 cells. The quadrupole MS analysis of the cellular lipids revealed an increased content of PC and TAG in EDL-expressing HepG2 cells compared with MUT-EDL-Ad-expressing and control cells. However, the MUT-EDL-Ad-expressing cells contained more PC than control cells. Additionally, EDL overexpression led to a 2-fold decrease in the amount of fatty acid synthase mRNA and, in turn, a slightly, but significantly, decreased rate of fatty acid (FA) synthesis in HepG2 cells. In the present study, we show for the first time that EDL efficiently supplies HepG2 cells with NEFA derived from HDL-PL, thus affecting cellular lipid composition and FA synthesis. PMID- 12553882 TI - Evolutionary connection between the catalytic subunits of DNA-dependent RNA polymerases and eukaryotic RNA-dependent RNA polymerases and the origin of RNA polymerases. AB - BACKGROUND: The eukaryotic RNA-dependent RNA polymerase (RDRP) is involved in the amplification of regulatory microRNAs during post-transcriptional gene silencing. This enzyme is highly conserved in most eukaryotes but is missing in archaea and bacteria. No evolutionary relationship between RDRP and other polymerases has been reported so far, hence the origin of this eukaryote-specific polymerase remains a mystery. RESULTS: Using extensive sequence profile searches, we identified bacteriophage homologs of the eukaryotic RDRP. The comparison of the eukaryotic RDRP and their homologs from bacteriophages led to the delineation of the conserved portion of these enzymes, which is predicted to harbor the catalytic site. Further, detailed sequence comparison, aided by examination of the crystal structure of the DNA-dependent RNA polymerase (DDRP), showed that the RDRP and the beta' subunit of DDRP (and its orthologs in archaea and eukaryotes) contain a conserved double-psi beta-barrel (DPBB) domain. This DPBB domain contains the signature motif DbDGD (b is a bulky residue), which is conserved in all RDRPs and DDRPs and contributes to catalysis via a coordinated divalent cation. Apart from the DPBB domain, no similarity was detected between RDRP and DDRP, which leaves open two scenarios for the origin of RDRP: i) RDRP evolved at the onset of the evolution of eukaryotes via a duplication of the DDRP beta' subunit followed by dramatic divergence that obliterated the sequence similarity outside the core catalytic domain and ii) the primordial RDRP, which consisted primarily of the DPBB domain, evolved from a common ancestor with the DDRP at a very early stage of evolution, during the RNA world era. The latter hypothesis implies that RDRP had been subsequently eliminated from cellular life forms and might have been reintroduced into the eukaryotic genomes through a bacteriophage. Sequence and structure analysis of the DDRP led to further insights into the evolution of RNA polymerases. In addition to the beta' subunit, beta subunit of DDRP also contains a DPBB domain, which is, however, distorted by large inserts and does not harbor a counterpart of the DbDGD motif. The DPBB domains of the two DDRP subunits together form the catalytic cleft, with the domain from the beta' subunit supplying the metal-coordinating DbDGD motif and the one from the beta subunit providing two lysine residues involved in catalysis. Given that the two DPBB domains of DDRP contribute completely different sets of active residues to the catalytic center, it is hypothesized that the ultimate ancestor of RNA polymerases functioned as a homodimer of a generic, RNA-binding DPBB domain. This ancestral protein probably did not have catalytic activity and served as a cofactor for a ribozyme RNA polymerase. Subsequent evolution of DDRP and RDRP involved accretion of distinct sets of additional domains. In the DDRPs, these included a RNA-binding Zn-ribbon, an AT-hook-like module and a sandwich-barrel hybrid motif (SBHM) domain. Further, lineage-specific accretion of SBHM domains and other, DDRP-specific domains is observed in bacterial DDRPs. In contrast, the orthologs of the beta' subunit in archaea and eukaryotes contains a four-stranded alpha + beta domain that is shared with the alpha-subunit of bacterial DDRP, eukaryotic DDRP subunit RBP11, translation factor eIF1 and type II topoisomerases. The additional domains of the RDRPs remain to be characterized. CONCLUSIONS: Eukaryotic RNA-dependent RNA polymerases share the catalytic double psi beta-barrel domain, containing a signature metal-coordinating motif, with the universally conserved beta' subunit of DNA-dependent RNA polymerases. Beyond this core catalytic domain, the two classes of RNA polymerases do not have common domains, suggesting early divergence from a common ancestor, with subsequent independent domain accretion. The beta-subunit of DDRP contains another, highly diverged DPBB domain. The presence of two distinct DPBB domains in two subunits of DDRP is compatible with the hypothesis that the ith the hypothesis that the ultimate ancestor of RNA polymerases was a RNA-binding DPBB domain that had no catalytic activity but rather functioned as a homodimeric cofactor for a ribozyme polymerase. PMID- 12553884 TI - Changes in the 5-HT2A receptor system in the pre-mammillary hypothalamus of the ewe are related to regulation of LH pulsatile secretion by an endogenous circannual rhythm. AB - BACKGROUND: We wanted to determine if changes in the expression of serotonin 2A receptor (5HT2A receptor) gene in the premammillary hypothalamus are associated with changes in reproductive neuroendocrine status. Thus, we compared 2 groups of ovariectomized-estradiol-treated ewes that expressed high vs low LH pulsatility in two different paradigms (2 groups per paradigm): (a) refractoriness (low LH secretion) or not (high LH secretion) to short days in pineal-intact Ile-de France ewes (RSD) and (b) endogenous circannual rhythm (ECR) in free-running pinealectomized Suffolk ewes in the active or inactive stage of their reproductive rhythm. RESULTS: In RSD ewes, density of 5HT2A receptor mRNA (by in situ hybridization) was significantly higher in the high LH group (25.3 +/- 1.4 vs 21.4 +/- 1.5 grains/neuron, P < 0.05) and 3H-Ketanserin binding (a specific radioligand) of the median part of the premammillary hypothalamus tended to be higher in the high group (29.1 +/- 4.0 vs 24.6 +/- 4.2 fmol/mg tissu-equivalent; P < 0.10). In ECR ewes, density of 5HT2A receptor mRNA and 3H-Ketanserin binding were both significantly higher in the high LH group (20.8 +/- 1.6 vs 17.0 +/- 1.5 grains/neuron, P < 0.01, and 19.7 +/- 5.0 vs 7.4 +/- 3.4 fmol/mg tissu equivalent; P < 0.05, respectively). CONCLUSIONS: We conclude that these higher 5HT2A receptor gene expression and binding activity of 5HT2A receptor in the premammillary hypothalamus are associated with stimulation of LH pulsatility expressed before the development of refractoriness to short days and prior to the decline of reproductive neuroendocrine activity during expression of the endogenous circannual rhythm. PMID- 12553885 TI - Characterization of the nodulation plasmid encoded chemoreceptor gene mcpG from Rhizobium leguminosarum. AB - BACKGROUND: In general, chemotaxis in Rhizobium has not been well characterized. Methyl accepting chemotaxis proteins are sensory proteins important in chemotaxis of numerous bacteria, but their involvement in Rhizobium chemotaxis is unclear and merits further investigation. RESULTS: A putative methyl accepting chemotaxis protein gene (mcpG) of Rhizobium leguminosarum VF39SM was isolated and characterized. The gene was found to reside on the nodulation plasmid, pRleVF39d. The predicted mcpG ORF displayed motifs common to known methyl-accepting chemotaxis proteins, such as two transmembrane domains and high homology to the conserved methylation and signaling domains of well-characterized MCPs. Phenotypic analysis of mcpG mutants using swarm plates did not identify ligands for this putative receptor. Additionally, gene knockouts of mcpG did not affect a mutant strain's ability to compete for nodulation with the wild type. Notably, mcpG was found to be plasmid-encoded in all strains of R. leguminosarum and R. etli examined, though it was found on the nodulation plasmid only in a minority of strains. CONCLUSIONS: Based on sequence homology R. leguminosarum mcpG gene codes for a methyl accepting chemotaxis protein. The gene is plasmid localized in numerous Rhizobium spp. Although localized to the sym plasmid of VF39SM mcpG does not appear to participate in early nodulation events. A ligand for McpG remains to be found. Apparent McpG orthologs appear in a diverse range of proteobacteria. Identification and characterization of mcpG adds to the family of mcp genes already identified in this organism. PMID- 12553886 TI - Genomic data sampling and its effect on classification performance assessment. AB - BACKGROUND: Supervised classification is fundamental in bioinformatics. Machine learning models, such as neural networks, have been applied to discover genes and expression patterns. This process is achieved by implementing training and test phases. In the training phase, a set of cases and their respective labels are used to build a classifier. During testing, the classifier is used to predict new cases. One approach to assessing its predictive quality is to estimate its accuracy during the test phase. Key limitations appear when dealing with small data samples. This paper investigates the effect of data sampling techniques on the assessment of neural network classifiers. RESULTS: Three data sampling techniques were studied: Cross-validation, leave-one-out, and bootstrap. These methods are designed to reduce the bias and variance of small-sample estimations. Two prediction problems based on small-sample sets were considered: Classification of microarray data originating from a leukemia study and from small, round blue-cell tumours. A third problem, the prediction of splice junctions, was analysed to perform comparisons. Different accuracy estimations were produced for each problem. The variations are accentuated in the small-data samples. The quality of the estimates depends on the number of train-test experiments and the amount of data used for training the networks. CONCLUSION: The predictive quality assessment of biomolecular data classifiers depends on the data size, sampling techniques and the number of train-test experiments. Conservative and optimistic accuracy estimations can be obtained by applying different methods. Guidelines are suggested to select a sampling technique according to the complexity of the prediction problem under consideration. PMID- 12553887 TI - Reporting degree of deacetylation values of chitosan: the influence of analytical methods. AB - PURPOSE: To investigate and compare the effect of three analytical methods, hydrogen bromide titrimetry (HBr titrimetry), infrared spectroscopy (IR spectroscopy), and first derivative UV-spectrophotometry (FDUV-spectrophotometry) in the determination of degree of deacetylation (DD) of chitosan. METHODS: Three different chitosan samples were selected for the DD quantification employing HBR titrimetry, IR spectroscopy with samples in the forms of KBr disc (at ratios of 1:2 and 1:3) and thin film (concentrations of 0.5 percent and 1 percent), and FDUV- spectrophotometry. RESULTS: The mean DD values of chitosan samples obtained by HBr titrimetry was significantly lower (p < 0.05) compared to IR spectroscopy using film (concentrations of 0.5 and 1 percent) as well as FDUV spectrophotometry (purified and unpurified samples), but were significantly higher (p< 0.005) than those obtained using IR spectroscopy using KBr disk except for Chit-S2 computed using baseline (a), which were closely comparable (p > 0.05). In the IR spectroscopic method, the DD values differed when computed using different baseline. It can be observed that the values calculated using baseline (b) was generally higher than those calculated using baseline (a). On the other hand, chitosan samples prepared in the form of film might not be significantly affected by the different baseline as no consistent trend was observed. For FDUV spectrophotometry, the DD values of purified Chit-S2 samples dried using oven and freeze dryer were comparable and not significantly different (p > 0.05). Similarly, no significant difference was observed for unpurified chitosan samples when dried using freeze dryer and oven (p > 0.05). Furthermore, the DD values of the purified and unpurified chitosan samples dried using freeze dryer were not significantly different except for Chit-S2. CONCLUSION: The DD values of chitosan were highly affected by the analytical methods employed. Hence, we proposed that the quantification method for DD should also be stated when reporting the DD value of chitosan sample. PMID- 12553888 TI - In-vitro studies of diclofenac sodium controlled-release from biopolymeric hydrophilic matrices. AB - PURPOSE: The objective of this study was to develop uncoated HPMC matrix tablets, evaluating the relationship and influence of different content levels of microcrystalline cellulose (MCC), starch, and lactose, in order to achieve a zero order release of Diclofenac Sodium. METHODS: HPMC matrix tablets of Diclofenac Sodium using microcrystalline cellulose (MCC), starch, and lactose were prepared by wet granulation process. The USP paddle method was selected to perform the dissolution profiles carried out in 900 mL 0.1 N HCl, and phosphate buffer. RESULTS: There was no significant difference in drug release between the hydrophilic matrices when the HPMC concentration was modified in low percentage. Release kinetics of Diclofenac Sodium from these swollen matrices was principally regulated by starch (17 percent) or lactose (17 percent), even on the presence of MCC. When starch (8.5 percent) and lactose (8.5 percent) were mixed at lower concentration in a ratio 1:1, MCC (5 percent or 7,5 percent) appeared to control the drug release. The release profile remained unchanged after three months storage of tablets. The best-fit release kinetics was achieved with the zero order plot, followed by the Higuchi and first-order equations. The data obtained proved that the formulations are useful for a sustained release of Diclofenac, due to the percentage released after 8 hours is nearly to 70 percent. CONCLUSIONS: The release of Diclofenac Sodium was influenced by the presence of MCC, and by the different concentrations of starch and lactose. Drug release kinetics from these formulations corresponded best to the zero-order kinetics. Compared to conventional tablets, release of the model drug from these HPMC matrix tablets was prolonged; as a result, an oral release dosage form to avoid the gastrointestinal adverse effects was achieved. PMID- 12553889 TI - Studies in topical application of niosomally entrapped Nimesulide. AB - PURPOSE: A niosome based transdermal drug delivery system of Nimesulide (NIM) was developed and extensively characterized and evaluated for in-vitro performance followed by in-vivo evaluation in rats by carrageenan induced rat paw edema method. METHOD: Niosomes were prepared by lipid film hydration technique using tweens and spans. Preparation of niosomes was optimized for highest percent drug entrapment (PDE). The prepared niosomes were incorporated into 1 percent carbopol gel base and the system was evaluated for drug diffusion across human cadaver skin (HCS) using modified validated diffusion cell. The drug retention studies in niosomes were performed at refrigerated temperature (2 degrees C - 8 degrees C) and at room temperature (25 degrees C+/-2 degrees C) for the period of 2 months. In-vivo performance of plain drug gel, niosomally-entrapped drug in carbopol gel base and marketed formulation were evaluated using acute rat paw edema method. RESULTS: Highest mean percentage edema inhibition (PEI) was observed for niosomal nimesulide gel after 24 hours i.e. 66.68 percent +/- 5.19 percent compared to plain drug gel i.e. 12.57 percent +/- 1.78 percent and marketed NIM formulation i.e. 20.49 percent +/- 0.91 percent. CONCLUSION: Findings of this investigation conclusively demonstrate prolongation of drug release and increase in amount of drug retention into the skin and permeation across the skin after niosomal encapsulation of NIM. Developed nimesulide niosomal gel formulation has also demonstrated enhanced anti-inflammatory activity compared to plain drug gel and marketed formulation. PMID- 12553890 TI - Inhibitory activity of Cinnamomum cassia bark-derived component against rat lens aldose reductase. AB - PURPOSE: To evaluate the inhibitory activity of active compounds isolated from Cinnamomum cassia bark against lens aldose reductase and compare to that of three commercially available compounds (cinnamyl alcohol, trans -cinnamic acid, and eugenol) and quercitrin as aldose reductase inhibitors. The IC (50) value of cinnamaldehyde was determined. METHODS: Active compound was purified on repeated silica gel column and HPLC (Waters Delta Prep 4000). Aldose reductase was prepared from lenses of Sprague-Dawley male rat eyes. The incubation mixture contained 135 mM Na, K-phosphate buffer (pH 7.0), 100 mM lithium sulfate, 0.03 mM NADPH, 0.04 mM DL-glyceraldehyde and 50 micro L of an enzyme preparation, with or without a plant extract. The reaction was initiated by adding NADPH at 37 degrees C and stopped by adding 0.5 N hydrochloric acid. Subsequently, 6 N NaOH containing 10 mM imidazole was added, and the mixture was incubated at 60 degrees C for 10 min to convert NADP into a fluorescent product. The fluorescence was measured with a spectrofluorophotometer. RESULTS: The biologically active constituents of C. cassia extract against lens aldose reductase were characterized as trans -cinnamaldehyde by spectral analysis. The IC (50) value of cinnamaldehyde is 0.003 mg/mL. However, cinnamyl alcohol, trans -cinnamic acid and eugenol exhibited only weak inhibition against aldose reductase. In comparison, quercitrin was 6 times more potent than cinnamaldehyde. CONCLUSIONS: These results suggest that cinnamaldehyde isolated from C. cassia barks may be useful as a lead compound and a medicinal foodstuff for aldose reductase inhibition. PMID- 12553891 TI - Fatty acid composition of Dracunculus vulgaris Schott (Araceae) seed oil from Turkey. AB - Dracunculus vulgaris Schott is only one taxon of the genus Dracunculus (Araceae) in Turkey. The tubers and the fruits with the seeds of D. vulgaris have long been in use for the treatment of rheumatism and hemorrhoids, respectively. The fatty acid composition of D. vulgaris seeds have been analyzed as their methyl esters by GC and GC-mass spectrometry. C16:0, C16:1n-7, C18:1n-9, C18:1n-7 (cis vaccenic acid), C18:2n-6 and 13-phenyl tridecanoic acids were found to be the main components in the seed oil. PMID- 12553892 TI - Formulation of enterosoluble microparticles for an acid labile protein. AB - PURPOSE: A microencapsulation method that preserves the activity of an acid labile protein was developed. METHODS: Solvent evaporation technique that employed ICH class 2 and 3 solvents methanol and acetone, respectively to dissolve pH-sensitive Eudragit polymers was investigated. Total protein released and lactase activities were measured using the USP method A for enteric cores and optimized with respect to process parameters. RESULTS: The percentage yields and entrapment efficiencies were directly proportional to solid content. The mean percentage yield and entrapment efficiency of selected sample was 84 +/- 0.9% and 88 +/- 0.7%, respectively. The residual specific activity of lactase in the selected sample was 89% +/- 0.8 with a net activity loss of 2 +/- 0.28% and 4 +/- 0.52% under ambient and stressed storage, respectively. Dibutyl sebacate levels, lower processing temperatures and lower processing speeds were influential in modulating enzyme activity. The most important formulation factor affecting lactase stability was Eudragit type, followed in decreasing order by processing temperature, processing speed, and solid percentage. CONCLUSIONS: Reliable control of lactase release was achieved by microencapsulating the enzyme with pH sensitive Eudragit L and S enteric polymers using either acetone- or methanol based solvent but lactase activity was preserved only in acetone-based formulations. PMID- 12553893 TI - Molecular modeling of potential new and selective PET radiotracers for the serotonin transporter. Positron Emission Tomography. AB - PURPOSE: Imaging the serotonin transporter (SERT) with Positron Emission Tomography (PET) provides a useful tool for understanding alterations of the serotonergic system. However, no optimal PET radiotracer for the SERT yet exists. The main purpose of this study was to design potential new and selective PET radiotracers for the SERT and to predict their binding affinity at both the SERT and the norepinephrine transporter. METHODS: Molecular Modeling was used for ligand design. Predictions of binding affinity were based on models generated by Comparative Molecular Field Analysis (CoMFA) and Comparative Molecular Similarity Indices Analysis (CoMSIA). RESULTS: A series of 100 compounds were suggested. As diphenyl sulfide derivatives like [(11)C] DASB have recently proven to be promising PET ligands, rational modification of the diphenyl sulfide scaffold has been performed. The novel compounds were predicted to be selective high affinity SERT ligands. Important new ideas are the introduction of a fluoroethyl oxycarbonyl group (ester) and a fluorethyl-carbonyl group (ketone), as well as a formyl group (aldehyde), and its corresponding oxime and imine. Another innovative suggestion is the replacement of the sulfur bridge with a cyanamide group and a fluoroethylamino group. CONCLUSIONS: The suggested compounds possess features providing new possibilities for carbon-11 or fluorine-18 labeling. Synthesis, biological testing, and screening for PET suitability are reasonable further steps. PMID- 12553894 TI - Molecular descriptor based on a molar refractivity partition using Randic-type graph-theoretical invariant. AB - PURPOSE: Development of a novel semi-empirical descriptor (MR(chi) for molecular modelling. METHOD: The index is based on a molar refractivity partition using Randictype graph-theoretical invariant. RESULTS: This hybrid index describes not only the London dispersive forces in a ligand fragment related to the molar refractivity but also structural features of the molecule It is also applicable in Quantitative Structure-Activity Relationship (QSAR) and Quantitative Structure Property Relationship (QSPR) studies. CONCLUSIONS: The method is convenient and can discriminate between isomers. PMID- 12553895 TI - Synthesis of isatin semicarbazones as novel anticonvulsants--role of hydrogen bonding. AB - PURPOSE: A series of substituted isatin semicarbazones and related bioisosteric hydrazones were designed and synthesised to meet the structural requirements essential for anticonvulsant properties. METHODS: The structures of all synthesised compounds were confirmed by means of infrared, proton magnetic resonance spectroscopy and by elemental analyses. All compounds were evaluated for their anticonvulsant activity by maximal electroshock (MES), subcutaneous metrazol (ScMet) and subcutaneous strychnine (ScSty) induced seizure methods and their neurotoxic effects were determined by rotorod test. RESULTS: A number of isatin semicarbazones exhibited significant protection after intraperitoneal administration at the dose of 100 and 300mg/kg. Some of them showed good anticonvulsant activity in MES test in rats after per oral administration at the dose of 30mg/kg. The bioisosteric hydrazone derivatives were inactive in all tests. Compound 6-chloroisatin-3- (4-bromophenyl)-semicarbazone has emerged as the most active analogue of the series showing good activity in all the three tests and was more active than phenytoin and valproic acid. CONCLUSIONS: The results evidenced the importance of hydrogen bonding and suggested a new pharmacophore model with four binding sites essential for anticonvulsant activity. PMID- 12553896 TI - Response surface optimization of the critical medium components for the production of alkaline protease by a newly isolated Bacillus sp. AB - PURPOSE: Optimization of the fermentation medium for maximum alkaline protease production was carried out using a new strain, Bacillus Sp. PE-11. METHODS: The carbon source (glucose), the nitrogen source (peptone) and salt solution were selected to optimize. A 2(3 )full factorial composite experimental design and response surface methodology were used in the design of experiments and in the analysis of results. This procedure limited the number of actual experiments performed while allowing for possible interactions between the three components. RESULTS AND DISCUSSION: The optimum values for the tested variables for the maximum alkaline protease production were; glucose 7.798 (g/L), peptone 9.548 (g/L) and salt solution 8.757%. The maximum alkaline protease production was 4,98,123 PU/L. This method was efficient; only 20 experiments were necessary to assess these conditions, and model adequacy was very satisfactory, as the coefficient of determination was 0.941. CONCLUSIONS: In the work, we have demonstrated the use of a central composite factorial design by determining the conditions leading to the high yield of enzyme production. Thus, smaller and less time consuming experimental designs could generally suffice for the optimization of many fermentation processes. PMID- 12553897 TI - Analysis of peptide and lipopeptide content in liposomes. AB - PURPOSE: To evaluate several methods for extraction of peptides from liposomal formulations as a first step in their quantification, and to determine the encapsulation efficiency for a panel of 8 peptides. METHODS: Eight peptides were chosen due to their importance in the field of vaccine development. Three different extraction media were examined: 25% ethanol, 98% ethanol, and 100% methanol. After extraction from liposomes, peptide content was measured using reverse phase HPLC. RESULTS: The effectiveness of the extraction media for peptide recovery varied considerably for the different peptides studied. In general, more hydrophilic peptides were recovered to a greater extent using 25% ethanol while more hydrophobic peptides were more thoroughly recovered using 98% ethanol. Encapsulation efficiencies (EE) ranged from 1% to 99% for the different peptides. No strong correlation was found between the average hydrophobicity values for the peptides and their EE. CONCLUSIONS: The most effective solvent for the extraction of a peptide from liposomes depends on the physicochemical properties of the peptide. Although the peptide sequence characteristics may provide guidance on the choice of the extraction media, only peptide recovery experiments will be able identify the optimal medium for extraction. PMID- 12553898 TI - Mathematical modelling of preparation of acyclovir liposomes: reverse phase evaporation method. AB - PURPOSE: The aim of this study was to derive simple reduced second order polynomial equation for constructing contour plots to obtain predetermined % drug entrapment (PDE) within liposomes of acyclovir (ACY) when prepared by reverse phase evaporation (REV) method using technique of three variables at three levels (3(3)) factorial design. METHOD: Three independent variables selected were volume of organic phase (x(1)), volume of aqueous phase (x(2)), and Drug/Phosphatidylcholine (PC) /Cholesterol (CHOL) in molar ratio (x(3)). Based on factorial design, twenty-seven batches of ACY liposomes were prepared by REV method. Prepared liposomal batches were evaluated for size, lamellarity, and PDE. The PDE (dependent variable) and the transformed values of independent variables were subjected to multiple regressions to establish a second order polynomial equation (full model). To simplify the equation, F-statistic was applied to reduce polynomial equation (reduced model) by neglecting nonsignificant (p>0.05) terms. The coefficient value for independent variable, Drug/PC/CHOL in molar ratio (x(3)) was found to be maximum (b(3) = 2.52) and hence the variable x(3) was considered to be a major contributing variable for PDE within liposomes by REV method. The reduced polynomial equation was used to plot three two dimensional contour plots at a fixed levels of -1, 0 and 1 of major contributing variable (x(3)) to obtain various combinations of values of two other independent variables (x(1) & x(2)) at predetermined PDE. The conformity of the established equation was checked by preparing three batches three times taking values of the independent variables from the contour plots for prefixed value of PDE. RESULTS: Prefixed PDE value taken for designing the experiment and results obtained experimentally were compared using student 't' test and difference between experimentally obtained and theoretically calculated values of PDE was found to be statistically nonsignificant (p>0.05). CONCLUSIONS: Findings of this study establishes the role of the derived equation and plotted contour plots in predicting the values of independent variables for preparation of ACY liposomes by REV method having predetermined PDE. PMID- 12553899 TI - Hydroxypropyl-beta-cyclodextrin-flutamide inclusion complex. II. Oral and intravenous pharmacokinetics of flutamide in the rat. AB - PURPOSE: The objective of this work was to determine the pharmacokinetics of flutamide (FLT) and its active metabolite, 2-hydroxy-flutamide (FLT-2-OH) in rats, following formulation in hydroxypropyl-Beta-cyclodextrin (FLT-HPBetaCyD). METHODS: The pharmacokinetics of FLT-HPBetaCyD, FLT-suspension (FLT-SUSP), and FLT-solution (FLT-COSOLV) were compared after oral (p.o.) and intravenous (i.v.) administration, respectively. In a non-crossover design, male Sprague-Dawley rats received each formulation as a single oral dose [15 mg (54 micro mol) FLT/kg] by oral gavage, or single i.v. dose [1.6 mg (5.8 micro mol) FLT/kg] via an indwelling jugular vein catheter. FLT and its metabolite, FLT-2-OH, were determined in plasma and urine aliquots by an HPLC method. RESULTS: In a preliminary in vitro experiment, using the dialysis bag dissolution method, 80% of a test dose of FLT was released from lyophilized FLT-HPBetaCyD into simulated gastric juice within 2 h, compared to less than 5% release from commercial FLT powder (FLT-SUSP). Following oral FLT-HPBetaCyD, the mean area under the plasma concentration curve (AUC(0- infinity)) for FLT, was 1580 +/- 228 ng x h/mL, with the maximum plasma concentration (Cmax; 1297 +/- 127 ng/mL) at 0.5 h (Tmax) after administration. The AUC(0- infinity) and C(max) were significantly higher than after FLT-SUSP (AUC(0- infinity) 748 +/- 206 ng x h/mL; C(max) 230 +/- 111 ng/mL and T(max) 2.33 +/- 0.29 h, respectively). After i.v. FLT-HPBetaCyD, the FLT AUC(0- infinity) was 1355 +/- 162 ng x h/mL, compared to 1421 +/- 283 ng x h/mL for FLT-COSOLV. FLT C(max) were 714 +/- 144 mL/h and 735 +/- 88 mL/h, respectively. The respective volumes of distribution (V(z)) were 369 +/- 191 mL and 242 +/- 25 mL. The plasma concentration-time profile and pharmacokinetic parameters of FLT after FLT-HPBetaCyD and FLT-COSOLV did not differ significantly. The pharmacokinetic parameters for FLT-2-OH were formulation independent after i.v. dosing, but AUC(0- infinity); C(max) and T(max), values were substantially greater with the FLT-HPBetaCyD in the oral study (40269 +/- 5875 ng x h/mL, 4062 +/- 502 ng/mL, and 3.50 +/- 0.41 h, respectively). CONCLUSIONS: FLT from FLT-HPBetaCyD was released rapidly into solution in vitro and in vivo. FLT-HPBetaCyD improved oral bioavailability relative to FLT-SUSP. Intravenous pharmacokinetic profiles for both FLT and FLT-2-OH were identical following either FLT-HPBetaCyD or FLT-COSOLV, indicating that the FLT-HPBetaCyD formulation behaved as a true solution. PMID- 12553900 TI - Endosome-specific localization and function of the ARF activator GNOM. AB - ARF GTPases act at multiple steps of the secretory and vacuolar/lysosomal trafficking pathways, but little is known about the spatial regulation of ARF activation. In this issue of Cell, Geldner et al. demonstrate that the Arabidopsis ARF activator GNOM localizes to endosomes where it controls the polarized trafficking of the auxin efflux carrier PIN1 to the basal plasma membrane. PMID- 12553901 TI - Ubiquitinating a phosphorylated Cdk inhibitor on the blades of the Cdc4 beta propeller. AB - Substrate binding by the SCFCdc4 ubiquitin ligase is regulated by phosphorylation. In this issue of Cell, Orlicky et al. describe the crystal structure of the Cdc4 subunit bound to a high-affinity substrate phosphopeptide. This structure provides insights into the binding interaction and how a precise mechanism involving multiple regulatory phosphorylations may be mediated by a single binding site. PMID- 12553902 TI - Visualizing genetic influences on human brain functions. AB - , in this issue of Cell, integrate genetics and functional brain imaging by showing that variation in the human brain-derived neurotrophic factor (BDNF) gene is associated with variation in episodic memory ability and in hippocampal neurochemistry and function. PMID- 12553903 TI - Sensing stretch is fundamental. AB - Stretch induces changes in cardiomyocyte biology that are implicated in heart failure, but the mechanism by which stretch is sensed and signals are transduced is unknown. New understanding of the Z disc elements of contractile units are beginning to elucidate the mechanism of stretch sensing and its relation to cardiac adaptation and disease. PMID- 12553904 TI - The ATRs, ATMs, and TORs are giant HEAT repeat proteins. PMID- 12553905 TI - From lineage to wiring specificity. POU domain transcription factors control precise connections of Drosophila olfactory projection neurons. AB - Axonal selection of synaptic partners is generally believed to determine wiring specificity in the nervous system. However, we have recently found evidence for specific dendritic targeting in the olfactory system of Drosophila: second order olfactory neurons (Projection Neurons) from the anterodorsal (adPN) and lateral (lPN) lineages send their dendrites to stereotypical, intercalating but non overlapping glomeruli. Here we show that POU domain transcription factors, Acj6 and Drifter, are expressed in adPNs and lPNs respectively, and are required for their dendritic targeting. Moreover, misexpression of Acj6 in lPNs, or Drifter in adPNs, results in dendritic targeting to glomeruli normally reserved for the other PN lineage. Thus, Acj6 and Drifter translate PN lineage information into distinct dendritic targeting specificity. Acj6 also controls stereotypical axon terminal arborization of PNs in a central target, suggesting that the connectivity of PN axons and dendrites in different brain centers is coordinately regulated. PMID- 12553906 TI - Twist regulates cytokine gene expression through a negative feedback loop that represses NF-kappaB activity. AB - During Drosophila embryogenesis, the dorsal transcription factor activates the expression of twist, a transcription factor required for mesoderm formation. We show here that the mammalian twist proteins, twist-1 and -2, are induced by a cytokine signaling pathway that requires the dorsal-related protein RelA, a member of the NF-kappaB family of transcription factors. Twist-1 and -2 repress cytokine gene expression through interaction with RelA. Mice homozygous for a twist-2 null allele or doubly heterozygous for twist-1 and -2 alleles show elevated expression of proinflammatory cytokines, resulting in perinatal death from cachexia. These findings reveal an evolutionarily conserved signaling circuit in which twist proteins regulate cytokine signaling by establishing a negative feedback loop that represses the NF-kappaB-dependent cytokine pathway. PMID- 12553907 TI - Liver tumor development. c-Jun antagonizes the proapoptotic activity of p53. AB - The transcription factor c-Jun mediates several cellular processes, including proliferation and survival, and is upregulated in many carcinomas. Liver-specific inactivation of c-Jun at different stages of tumor development was used to study its role in chemically induced hepatocellular carcinomas (HCCs) in mice. The requirement for c-jun was restricted to early stages of tumor development, and the number and size of hepatic tumors was dramatically reduced when c-jun was inactivated after the tumor had initiated. The impaired tumor development correlated with increased levels of p53 and its target gene noxa, resulting in the induction of apoptosis without affecting cell proliferation. Primary hepatocytes lacking c-Jun showed increased sensitivity to TNF-alpha-induced apoptosis, which was abrogated in the absence of p53. These data indicate that c Jun prevents apoptosis by antagonizing p53 activity, illustrating a mechanism that might contribute to the early stages of human HCC development. PMID- 12553908 TI - X-ray structures of Myc-Max and Mad-Max recognizing DNA. Molecular bases of regulation by proto-oncogenic transcription factors. AB - X-ray structures of the basic/helix-loop-helix/leucine zipper (bHLHZ) domains of Myc-Max and Mad-Max heterodimers bound to their common DNA target (Enhancer or E box hexanucleotide, 5'-CACGTG-3') have been determined at 1.9 A and 2.0 A resolution, respectively. E box recognition by these two structurally similar transcription factor pairs determines whether a cell will divide and proliferate (Myc-Max) or differentiate and become quiescent (Mad-Max). Deregulation of Myc has been implicated in the development of many human cancers, including Burkitt's lymphoma, neuroblastomas, and small cell lung cancers. Both quasisymmetric heterodimers resemble the symmetric Max homodimer, albeit with marked structural differences in the coiled-coil leucine zipper regions that explain preferential homo- and heteromeric dimerization of these three evolutionarily related DNA binding proteins. The Myc-Max heterodimer, but not its Mad-Max counterpart, dimerizes to form a bivalent heterotetramer, which explains how Myc can upregulate expression of genes with promoters bearing widely separated E boxes. PMID- 12553909 TI - Schizosaccharomyces pombe Int6 and Ras homologs regulate cell division and mitotic fidelity via the proteasome. AB - Yin6 is a yeast homolog of Int6, which is implicated in tumorigenesis. We show that Yin6 binds to and regulates proteasome activity. Overexpression of Yin6 strengthens proteasome function while inactivation weakens and causes the accumulation of polyubiquitinated proteins including securin/Cut2 and cyclin/Cdc13. Yin6 regulates the proteasome by preferentially interacting with Rpn5, a conserved proteasome subunit, and affecting its localization/assembly. We showed previously that Yin6 cooperates with Ras1 to mediate chromosome segregation; here, we demonstrate that Ras1 similarly regulates the proteasome via Rpn5. In yeast, human Int6 binds Rpn5 and regulates its localization. We propose that human Int6, either alone or cooperatively with Ras, influences proteasome activities via Rpn5. Inactivating Int6 can lead to accumulation of mitotic regulators affecting cell division and mitotic fidelity. PMID- 12553910 TI - The Arabidopsis GNOM ARF-GEF mediates endosomal recycling, auxin transport, and auxin-dependent plant growth. AB - Exchange factors for ARF GTPases (ARF-GEFs) regulate vesicle trafficking in a variety of organisms. The Arabidopsis protein GNOM is a brefeldin A (BFA) sensitive ARF-GEF that is required for the proper polar localization of PIN1, a candidate transporter of the plant hormone auxin. Mutations in GNOM lead to developmental defects that resemble those caused by interfering with auxin transport. Both PIN1 localization and auxin transport are also sensitive to BFA. In this paper, we show that GNOM localizes to endosomes and is required for their structural integrity. We engineered a BFA-resistant version of GNOM. In plants harboring this fully functional GNOM variant, PIN1 localization and auxin transport are no longer sensitive to BFA, while trafficking of other proteins is still affected by the drug. Our results demonstrate that GNOM is required for the recycling of auxin transport components and suggest that ARF-GEFs regulate specific endosomal trafficking pathways. PMID- 12553911 TI - Energetics of protein transport across biological membranes. a study of the thylakoid DeltapH-dependent/cpTat pathway. AB - Among the pathways for protein translocation across biological membranes, the DeltapH-dependent/Tat system is unusual in its sole reliance upon the transmembrane pH gradient to drive protein transport. The free energy cost of protein translocation via the chloro-plast DeltapH-dependent/Tat pathway was measured by conducting in vitro transport assays with isolated thylakoids while concurrently monitoring energetic parameters. These experiments revealed a substrate-specific energetic barrier to cpTat-mediated transport as well as direct utilization of protons from the gradient, consistent with a H+/protein antiporter mechanism. The magnitude of proton flux was assayed by four independent approaches and averaged 7.9 x 10(4) protons released from the gradient per transported protein. This corresponds to a DeltaG transport of 6.9 x 10(5) kJ.mol protein translocated(-1), representing the utilization of an energetic equivalent of 10(4) molecules of ATP. At this cost, we estimate that the DeltapH-dependent/cpTat pathway utilizes approximately 3% of the total energy output of the chloroplast. PMID- 12553912 TI - Structural basis for phosphodependent substrate selection and orientation by the SCFCdc4 ubiquitin ligase. AB - Cell cycle progression depends on precise elimination of cyclins and cyclin dependent kinase (CDK) inhibitors by the ubiquitin system. Elimination of the CDK inhibitor Sic1 by the SCFCdc4 ubiquitin ligase at the onset of S phase requires phosphorylation of Sic1 on at least six of its nine Cdc4-phosphodegron (CPD) sites. A 2.7 A X-ray crystal structure of a Skp1-Cdc4 complex bound to a high affinity CPD phosphopeptide from human cyclin E reveals a core CPD motif, Leu-Leu pThr-Pro, bound to an eight-bladed WD40 propeller domain in Cdc4. The low affinity of each CPD motif in Sic1 reflects structural discordance with one or more elements of the Cdc4 binding site. Reengineering of Cdc4 to reduce selection against Sic1 sequences allows ubiquitination of lower phosphorylated forms of Sic1. These features account for the observed phosphorylation threshold in Sic1 recognition and suggest an equilibrium binding mode between a single receptor site in Cdc4 and multiple low-affinity CPD sites in Sic1. PMID- 12553913 TI - The BDNF val66met polymorphism affects activity-dependent secretion of BDNF and human memory and hippocampal function. AB - Brain-derived neurotrophic factor (BDNF) modulates hippocampal plasticity and hippocampal-dependent memory in cell models and in animals. We examined the effects of a valine (val) to methionine (met) substitution in the 5' pro-region of the human BDNF protein. In human subjects, the met allele was associated with poorer episodic memory, abnormal hippocampal activation assayed with fMRI, and lower hippocampal n-acetyl aspartate (NAA), assayed with MRI spectroscopy. Neurons transfected with met-BDNF-GFP showed lower depolarization-induced secretion, while constitutive secretion was unchanged. Furthermore, met-BDNF-GFP failed to localize to secretory granules or synapses. These results demonstrate a role for BDNF and its val/met polymorphism in human memory and hippocampal function and suggest val/met exerts these effects by impacting intracellular trafficking and activity-dependent secretion of BDNF. PMID- 12553914 TI - Two-photon calcium imaging reveals an odor-evoked map of activity in the fly brain. AB - An understanding of the logic of odor perception requires a functional analysis of odor-evoked patterns of activity in neural assemblies in the brain. We have developed a sensitive imaging system in the Drosophila brain that couples two photon microscopy with the specific expression of the calcium-sensitive fluorescent protein, G-CaMP. At natural odor concentration, each odor elicits a distinct and sparse spatial pattern of activity in the antennal lobe that is conserved in different flies. Patterns of glomerular activity are similar upon imaging of sensory and projection neurons, suggesting the faithful transmission of sensory input to higher brain centers. Finally, we demonstrate that the response pattern of a given glomerulus is a function of the specificity of a single odorant receptor. The development of this imaging system affords an opportunity to monitor activity in defined neurons throughout the fly brain with high sensitivity and excellent spatial resolution. PMID- 12553915 TI - Infants' learning, memory, and generalization of learning for bimodal events. AB - Study 1 investigated whether infants 3 and 7 months of age show differential learning of and memory for sight-sound pairs depending on whether or not temporal synchrony was present; memory was assessed after a 10-min and 1-week interval. Study 2 examined whether 7-month-olds show generalization of learning when they encounter novel bimodal events that are similar (changes in size, orientation, or color, and spectral sound properties) to the sight-sound pairs learned 1 week earlier based on temporal synchrony. For Study 1, infants received a familiarization phase followed by a paired-comparison preference procedure to assess for learning of the sight-sound pairs. One week later a memory test was given. Results confirmed that 7-month-olds had no difficulty learning auditory visual pairings regardless of whether or not events were temporally synchronous, and they remembered these 10 min and 1 week later. In contrast, 3-month-olds showed poorer learning of sight-sound associations in the no-synchrony than synchrony conditions, and memory for sight-sound pairs 1 week later was shown only for the synchrony conditions. Results for Study 2 revealed generalization of learning of bimodal pairings under all stimulus conditions after a 1-week interval at 7 months of age. Implications of these findings for development of intersensory knowledge are discussed. PMID- 12553916 TI - Developmental changes in the processing of hierarchical shapes continue into adolescence. AB - The present study was designed to trace the normal development of local and global processing of hierarchical visual forms. We presented pairs of hierarchical shapes to children and adults and asked them to indicate whether the two shapes were the same or different at either the global or the local level. In Experiments 1 (6-year-olds, 10-year-olds, adults) and 2 (10-year-olds, 14-year olds, adults), we presented stimuli centrally. All age groups responded faster on global trials than local trials (global precedence effect), but the bias was stronger in children and diminished to the adult level between 10 and 14 years of age. In Experiment 3 (10-year-olds, 14-year-olds, adults), we presented stimuli in the left or right visual field so that they were transmitted first to the contralateral hemisphere. All age groups responded faster on local trials when stimuli were presented in the right visual field (left hemisphere); reaction times on global trials were independent of visual field. The results of Experiment 3 suggest that by 10 years of age the hemispheres have adult-like specialization for the processing of hierarchical shapes, at least when attention is directed to the global versus local level. Nevertheless, their greater bias in Experiments 1 and 2 suggests that 10-year-olds are less able than adults to modulate attention to the output from local versus global channels-perhaps because they are less able to ignore distractors and perhaps because the cerebral hemispheres are less able to engage in parallel processing. PMID- 12553917 TI - Differential memory-preserving effects of reminders at 6 months. AB - Although reactivation and reinstatement reminders differ procedurally, differences in their memory-preserving effects have been described as artifactual. In three experiments, we examined this conclusion. One hundred and twelve 6-month-olds learned an operant task, forgot it, received a reactivation or reinstatement reminder to recover the inactive memory, and were tested after increasing delays until they forgot it again. In Experiments 1a and 1b, a single reactivation reminder extended infants' memory of an operant mobile task for 2 weeks after reminding, but a single reinstatement extended it for 4 weeks, when testing was discontinued. In Experiment 2, a single reinstatement extended 6 month-olds' memory of an operant train task for 19 weeks after reminding, when infants were almost 1 year old. After reactivation, infants remember this task for only 2 weeks. The finding that the memory-preserving effect of reinstatement is greater by an order of magnitude suggests that procedural differences between the two reminders have functional significance. PMID- 12553918 TI - Time and number discrimination in a bisection task with a sequence of stimuli: a developmental approach. AB - Children, aged 5 and 8 years, and adults were tested in a bisection task with a sequence of stimuli in which time and number co-varied. In a counting and a non counting condition, they were instructed either to process the duration of this sequence while ignoring the number of stimuli (temporal bisection), or to process the number of stimuli while ignoring the duration (numerical bisection). In the temporal bisection task, number interfered with the 5-year-olds' temporal performance, indicating that young children did not process time and number independently in a sequence of stimuli when they had to attend to duration. However, number interference decreased both with age and counting strategy. In contrast, in the numerical bisection task, duration did not interfere with numerical discrimination for any age group. PMID- 12553920 TI - Scanning electron microscopic observations of 'fractured' biodegradable plates and screws. AB - BACKGROUND: We encountered two out of 100 cases in which implanted biodegradable plates and screws had fractured within 1 month postoperatively. MATERIAL: Failure of the material was confirmed through clinical symptoms, radiographs or CT findings. In addition, four specimens obtained from these two cases were examined with regard to their ultrastructure using scanning electron microscopy. RESULTS: Several principal patterns of the fractured surface were found: (1) gradual cracking, i.e. 'circular stair' and, (2) tortuous threads, i.e. a wavy line. It is conceivable that the material may not have been hit by major sudden forces but a disproportion between the thread configuration and the drilled hole may have led to screw loosening and torsion. Subsequently, the threads were deformed in a 'wavy' manner, finally leading to cracking and fracture of plates and screws. Fractures of plates and screws due to these instabilities are thought to be distinguishable from material resorption. CONCLUSION: In the application of biodegradable materials, more than two screws per single bone segment should be used as a principle of plate-fixation technique in order to avoid a stability compromising situation, particularly in the stress-bearing areas of the maxillofacial region. Moreover, three-dimensional fixation using more than two plates is recommended in the facial skeleton e.g. zygomatic tripod. Intermaxillary fixation should also be considered to reinforce initial stability in stress-bearing areas. PMID- 12553919 TI - Jaw malformations plus displacement and numerical aberrations of teeth in neurofibromatosis type 1: a descriptive analysis of 48 patients based on panoramic radiographs and oral findings. AB - AIM: The aim of this study was to analyse jaw malformations and tooth displacement in patients with neurofibromatosis type 1 (NF1). MATERIAL AND METHODS: Forty-eight patients were included in the study (male or female 24 each). All fulfilled the current NIH diagnostic criteria for NF1. The age range was 2.5-66 years. The type of neurofibroma was histologically proven in surgically treated patients. Patients with disseminated cutaneous neurofibromata and those with the plexiform type were distinguished. The analysis was based on physical investigation, photographs, panoramic radiographs and dental casts. RESULTS: With the emphasis on alterations of tooth position, deformities of the adjacent bones and malocclusion, the majority of these patients (26) were affected by plexiform neurofibromata. In the other 22 patients with disseminated neurofibromata, malformations of the alveolar ridge were absent and individual oral symptoms were rarely found and were mild, and in all cases were unimpaired. Numerical aberrations and retention of molars was exclusively associated with a trigeminal nerve affected by plexiform neurofibroma. Aplasia of a second lower molar was recognized in four of these plexiform-neurofibroma patients. CONCLUSION: It is widely accepted that malformations of the facial skeleton are often of genetic origin. However, in this study these malformations were strongly associated with plexiform neurofibromata originating from the trigeminal nerve. Thus, in addition to presently unknown genetic factors, the pattern of skeletal malformation can be caused by tumour invasion and local destruction, e.g. the neuromuscular unit or prenatal development of the plexiform neurofibroma in the inferior alveolar nerve. It is further concluded that epidemiologic studies on the incidence and severity of NF1 in the oral and maxillofacial region have to distinguish between patients with or without plexiform neurofibroma, especially when analysing alterations and deformities of the jaws, teeth and malocclusion. Aplasia of second inferior molars is an additional (dental) finding associated with plexiform neurofibromata in NF1. PMID- 12553921 TI - Intracranial migration of fixation wires following correction of craniosynostosis in an infant. AB - BACKGROUND: In the craniofacial skeleton, osseous fixation techniques for stabilization include stainless steel wire, miniplates or bioresorbable plates; in some cases, stainless steel wires are still indicated. Most recently, several case reports have demonstrated that microplates or stainless steel wires migrate intracranially when used in the craniofacial skeleton in neonates. PATIENT: We present the case of a 7-year-old male who underwent treatment of brachycephaly at the age of 5 months. In this patient, internal migration of wires was observed. CONCLUSION: In our opinion, there is a difference between migration of wires and of plates in the growing cranium. Factors affecting the incidence of material migration in cranioplasty are believed to include (1) age, (2) site and (3) the material itself. PMID- 12553922 TI - Differences in cephalometric and occlusal outcome of cleft palate patients regarding different surgical techniques. AB - INTRODUCTION: The purpose of this study was to assess differences of the long term results following surgical treatment in patients with cleft palate treated by two different surgical concepts. PATIENTS: Fifty-nine adult patients operated on for cleft palate were examined. Thirty palates were closed by a two stage (Widmaier and Veau) and 29 by a single-stage procedure (Veau's pedicled flap). METHODS: Lateral cephalometric and model analysis was performed. In the cephalometric analysis, the vertical and horizontal parameters of the position of maxilla and midface and transverse and sagittal dimensions of the models were compared between the two groups. RESULTS: Model analysis: According to the Bolton analysis the maxillary dental arch was too large in 22 patients in each group. The other patients had mandibular arches that were too large. In 18 patients with two-stage closure and in 9 patients with one-stage closure, a space deficit in the lateral part of the maxilla was observed. Persisting transverse deficits were seen in all patients with two-stage repairs and in 11 patients with one-stage repairs. The deficit was more severe in the molar area in the first group and almost equally severe in the premolar and the molar regions of the second group. A sagittal deficiency was found more often in patients with two-stage repairs while Angle's class I occlusion was seen more often in patients with one-stage surgery. Lateral cephalometry: Similar SNA-angles were seen in both groups whereas the ANB-angle was greater following two-stage repair. In both patient groups a low inclination of the midface was seen. The vertical dimension of the midface in comparison with the lower face was normal in the one-stage group; in the other group a deficiency of the anterior midface height was registered. CONCLUSION: There was a more severe growth impairment of the midface in patients with this type of two-stage palatal repair. The horizontal deficiency was similar in both groups. The long-term occlusal result revealed smaller sagittal and transverse deficiencies in patients with this type of single-stage closure. PMID- 12553923 TI - Bone regeneration following injection of mesenchymal stem cells and fibrin glue with a biodegradable scaffold. AB - AIM: The purpose of this study was to determine whether a combination of fibrin glue, beta-tricalcium phosphate as a biodegradable (beta-TCP) and mesenchymal stem cells would provide three-dimensional templates for bone growth resulting in new bone formation at heterotopic sites in the rat with plasticity. MATERIAL AND METHODS: Growing stem cells and developing matrices, explanted from the rat femur, were fragmented and mixed with fibrin glue in a syringe. The cells/beta TCP fibrin glue admixtures were injected into the subcutaneous space on the dorsum of the rat. RESULTS: Eight weeks after implantation, gross morphology revealed a pearly opalescence and firm consistency. Histological inspections showed newly formed bone structures in all admixtures, but none in the control groups when only fibrin glue and beta-TCP were injected. Osteopontin, a protein important in bone development, was identified by using antibodies in all cells/beta-TCP fibrin glue admixtures. CONCLUSION: Mesenchymal stem cells/beta TCP fibrin glue admixtures can result in successful bone formation. This technique holds the promise of a minimally invasive means of generating autogenous bone to correct or reconstruct bony defects. PMID- 12553924 TI - Making bone: implant insertion into tissue-engineered bone for maxillary sinus floor augmentation-a preliminary report. AB - Autologous, allogenic and alloplastic materials for bony reconstruction in the cranio-maxillofacial area have many drawbacks thus stimulating the on-going search for new (bio-)materials. Whereas cultivated skin and mucosa are already in clinical routine use in head and neck reconstruction, so far there has been no successful clinical application to the best of our knowledge of periosteum derived, tissue-engineered bone for augmentation of the edentulous posterior maxilla. In a pilot study, augmentation of the posterior maxilla was carried out using a bone matrix derived from mandibular periosteal cells on a polymer fleece. This paper demonstrates fabrication of the matrix, clinical application, and the histological results in two patients. The results suggest that periosteum-derived osteoblasts on a suitable matrix form lamellar bone within 4 months which allows reliable implant insertion. PMID- 12553925 TI - A sample method of measuring bone graft volume, technical note. AB - A simple method of measuring the volume of cortico-cancellous bone grafts is described. The method is quick and reliable. Only materials readily available in the operation theatre are used. PMID- 12553926 TI - Cost, operation and hospitalization times in distraction osteogenesis versus sagittal split osteotomy. AB - INTRODUCTION: Distraction osteogenesis in 'common' surgical orthodontics is mentioned as an alternative for conventional sagittal split osteotomy. After a 'learning curve' in the surgical skills of distraction, the two techniques can be compared concerning time and cost aspects. PATIENTS: Forty-seven patients (male n=28, female n=19, age 11.7-17.9 yr (mean 14.2) with an Angle's class II division 1malocclusion of skeletal mandibular origin were operated on using distraction osteogenesis and were compared with a second group of 21 patients (male n=4, female n=17, age 16-36yr (mean 22.8) who underwent bilateral sagittal split osteotomies in the same period in order to correct the same type of dysgnathia. METHOD: The first group of 47 patients was treated with intraorally placed bone born distraction devices to correct mandibular deficiency following a standard protocol, with removal of the third molar tooth germs if present. The second group of 21 patients underwent standard sagittal split osteotomies to correct the mandibular deficiency. The costs of hospitalization, distraction devices and operation time were compared. RESULTS: In this study, distraction osteogenesis took on average more operative time (mean 37%), but 1 day less hospitalization than the bilateral sagittal split osteotomies. The surgical cost of distraction osteogenesis was 36% higher than the conventional sagittal split osteotomy. CONCLUSION: In this study, correcting Angle's class II deficiencies by means of distraction osteogenesis was shown not to be a time-saving procedure when compared with sagittal split osteotomy. Surgical costs were significantly higher using distraction, mainly due to the price of the distractors. Changes in hospital policies concerning hospital admission of adults and children and European legislation concerning re-usability of surgical instruments may balance the cost of both procedures. PMID- 12553927 TI - Depression, anxiety and quality of life: outcome 9 months after facial cosmetic surgery. AB - OBJECTIVE: The request for cosmetic surgery is of a psychological nature. Very few studies have quantitatively assessed whether or not this psychological need was actually satisfied, and more precisely, which psychic components were satisfied. MATERIAL AND METHODS: This is a multicentric, prospective cohort study. One hundred and three patients scheduled for facial cosmetic surgery from three different hospitals were examined before and after surgery using four assessment scales validated using European populations. The Montgomery and Asberg depression rating scale (MADRS) measured the existence and intensity of depression, the self-assessment test of thoughts in social interaction (SISST) measured the positive or inhibitory thoughts in the context of social relationships, and the European quality of life 5 dimensions (EQ-5D) (generic test) measured the quality of life. In addition, a semi-directive interview was specially created by our team. For statistical analysis, ANOVA and Student's t test were applied. RESULTS: Twenty-four patients were lost to follow-up. Although the initial MADRS index was high (p<0.05), it did not change after surgery (p>0.1). SISST+ (positive thoughts): the social anxiety of the individual examined was significantly greater than that of the control group (p<0.005) and improved after surgery (p<0.01). The SISST- (inhibitory thoughts) did not change (p>0.1). The EQ-5D visual analogue scale (VAS) did not reveal any difference (p>0.1) while the descriptive EQ-5D demonstrated over-representation of anxiety/depression (p<0.01), and an improvement of this (p<0.05) postoperatively. The mean subjective satisfaction index was 8.1 (scale of 1-10) without sharing any influence of the complications suffered (65% of the patients made self assessments). CONCLUSION: The best indications for facial cosmetic surgery seem to be a lack of self-confidence associated with a desire for social interaction, and a request focused on a specific physical feature. The results presented add documentary confirmation to the impression shared by the majority of cosmetic surgeons. However, it was also confirmed that cosmetic surgery is not limited to its technical components, but remains a medical act which must consider the overall effect on the whole patient. PMID- 12553928 TI - Cranio-maxillofacial trauma: a 10 year review of 9,543 cases with 21,067 injuries. AB - INTRODUCTION: Cranio-maxillofacial trauma management requires pertinent documentation. Using a large computerized database, injury surveillance and research data describe the whole spectrum of injuries. The goal of this study was to assess the effect of the five main causes of accidents resulting in facial injury on the severity of cranio-maxillofacial trauma. PATIENTS AND METHODS: During a period of 10 years (1991-2000) 9,543 patients were admitted to the Department of Oral and Maxillofacial Surgery, University Hospital of Innsbruck with cranio-maxillofacial trauma. Data of patients were prospectively recorded including cause of injury, age and gender, type of injury, injury mechanisms, location and frequency of soft tissue injuries, dentoalveolar trauma, facial bone fractures and concomitant injuries. Statistical analyses performed included descriptive analysis, chi square test, Fisher's exact test, and Mann-Whitney's U test. This was followed by logistic regression analyses for the three injury types to determine the impact of the five main causes on the type of injury at different ages in facial trauma patients. RESULTS: Five major categories/mechanisms of injury existed: in 3,613 (38%) cases it was activity of daily life, in 2991 (31%) sports, 1170 (12%) violence, in 1,116 (12%) traffic accidents, in 504 (5%) work accidents and in 149 (2%) other causes. A total of 3,578 patients (37.5%) had 7,061 facial bone fractures, 4,763 patients (49.9%) suffered from 6,237 dentoalveolar, and 5,968 patients (62.5%) from 7,769 soft tissue injuries. Gender distribution showed an overall male-to-female ratio of 2.1 to 1 and the mean age was 25.8+/-19.9 years; but both varied greatly depending on the injury mechanism (facial bone fractures: 35.4+/-19.5 years, higher risk for males; soft tissue injuries: 28.7+/-20.5, no gender preference; dentoalveolar trauma: 18+/-15.6, elevated risk for females). For patients sustaining facial trauma, logistic regression analyses revealed increased risks for facial bone fractures (225%), soft tissue lesions (58%) in patients involved in traffic accidents, and dental trauma (49%) during activities of daily life and play accidents. When compared with other causes, the probability of suffering soft tissue injuries and dental trauma, but not facial bone fractures, is higher in sports-related accidents, 12 and 16%, respectively. CONCLUSION: This study differentiated between injury mechanisms in cranio-maxillofacial trauma. The specially trained surgeons treating cranio-maxillofacial trauma are the primary source of information for the public and legislators on implementing preventive measures for high-risk activities. In facial trauma, older persons are prone to bone fractures (increase of 4.4%/year of age) and soft tissue injuries (increase of 2%/year of age) while younger persons are more susceptible to dentoalveolar trauma (decrease of 4.5%/year of age). PMID- 12553929 TI - Synchronous tumours of the unilateral parotid gland: rare or undetected? AB - INTRODUCTION: Multiple tumours of the parotid gland with the same histological appearance may occur as synchronous unilateral tumours, but bilaterality has also been reported. Synchronous multiple unilateral parotid tumours with different histology remain rare. METHODS: Between January 1988 and May 2002, a total of 341 patients underwent parotidectomy in our department. Medical charts were reviewed retrospectively for synchronous multiple unilateral tumours. RESULTS: Fourteen patients had two or more tumours within the same specimen. The combinations encountered were two to four adenolymphomas (n=9), adenolymphoma plus pleomorphic adenoma (n=3), adenolymphoma plus MALT lymphoma (n=1), and pleomorphic adenoma plus acinic cell carcinoma (n=1). The outcome was clinical freedom from signs of tumour recurrence in any patient (mean follow-up = 51 months). CONCLUSION: Synchronous multiple unilateral parotid tumours usually include two or more adenolymphomas and might occur more often than previously realized. The possibility of a concomitant carcinoma, and the prevention of recurrent tumours, may warrant a more radical surgical excision of the parotid gland, accurate intraoperative examination of the resected specimen, and routine histological evaluation of the entire specimen. Preoperative radiological investigation may further increase the chances of finding multiple parotid tumours. PMID- 12553931 TI - Sequence length required for homologous recombination in Cryptococcus neoformans. AB - Cryptococcosis is a major threat to immunocompromised individuals. Isolates of Cryptococcus neoformans var. grubii and var. neoformans are responsible for most of the infections in the United States and Europe. In depth analysis of the virulence phenotype of this organism requires the generation of specific gene disruptions. The minimum sequence requirements for efficient homologous recombination has not been determined in Cryptococcus. To investigate the flanking DNA length requirements for efficient homologous recombination in variety grubii, the rates of homologous recombination of constructs with different lengths of flanking sequence at two loci, CAP59 and CNLAC1, were examined. Five gene disruption constructs were prepared for each locus with symmetric lengths of sequence homologous to the target gene with approximately 50, 100, 200, 300 or 400bp flanking the selectable marker for hygromycin resistance. In addition, two asymmetric constructs with 50bp on one side and 400bp on the other side were generated for each locus. Overall, symmetric constructs with 300bp or more of flanking sequence on each side and the asymmetric constructs were efficiently targeted for gene disruption by homologous recombination in C. neoformans var. grubii. With one exception, the rate of recovery of homologous recombinants using the longer or asymmetric constructs as targeting vectors was greater than five percent of total transformants. Symmetrical constructs with 100bp or less of homologous flanking sequence did not efficiently generate targeted gene disruptions because the rate of homologous recombinants was less than or equal to 1%. PMID- 12553932 TI - Tandem cytochrome P450 monooxygenase genes and splice variants in the white rot fungus Phanerochaete chrysosporium: cloning, sequence analysis, and regulation of differential expression. AB - Phanerochaete chrysosporium, the model white rot fungus, uses P450 monooxygenation for oxidation of xenobiotic chemicals. To characterize its P450 systems, we have isolated two tandemly linked full-length P450 monooxygenase genes, pc-1 (2510bp) and pc-2 (2503bp). cDNA sequence and splice site analysis showed multiple small putative introns (35-82bp), 17 in pc-1 and 16 in pc-2. The deduced P450 proteins PC-1 (531 aa, MW 60,070) and PC-2 (538 aa, MW 61,765) showed 55% aa identity with each other and low relatedness (25-36%) with P450 gene families CYP52, CYP86, CYP4, and CYP3 decreasing in that order. Alternate splicing was identified for pc-1 transcription. Real-time quantitative RT-PCR analysis on 4-day cultures showed an elevated level of expression ( approximately 8-fold) of pc-1 in nutrient-limited (ligninolytic) as compared to nutrient-rich (non-ligninolytic) conditions. Interestingly, the pc-1 gene is expressible in both primary and secondary phases of growth with peak expression occurring with secondary metabolism. PMID- 12553933 TI - Histone deacetylase Hda1 acts as repressor of the Ustilago maydis biotrophic marker gene mig1. AB - The Ustilago maydis mig1 gene is extensively up-regulated during growth within its host plant. A genetic approach was set up to identify mutants expressing mig1 during axenic growth. Five independent mutants were identified that not only displayed increased transcript levels of mig1 but also of egl1, an endoglucanase expressed in dikaryotic filaments. egl1 has recently been shown to be repressed by Hda1, a putative histone deacetylase [Reichmann et al., submitted]. The identified UV mutants shared other phenotypes with hda1 deletion mutants like enhanced pigmentation and the inability to produce teliospores in maize tumours. Complementation and sequence analysis demonstrated that all five UV mutants contained point mutations in the hda1 gene. Despite a common repression mechanism, expression levels of mig1 and egl1 were significantly different during axenic and biotrophic growth, providing evidence for additional regulatory inputs from the respective growth stage. Furthermore, while egl1 is subject to repression by the U. maydis regulator Rum1, this was not the case for mig1. U. maydis strains deleted in either hda1 or rum1 were not affected in mig1 expression in the tumour stage. Transcript levels conferred by mig1 promoters deleted in negatively cis-acting sequences exceeded those in hda1 mutants, suggesting additional negative factors governing mig1 expression. PMID- 12553934 TI - A genetic interval and physical contig spanning the Peronospora parasitica (At) avirulence gene locus ATR1Nd. AB - In Peronospora parasitica (At) (downy mildew), the genetic determinants of cultivar-specific recognition by Arabidopsis thaliana are the Arabidopsis thaliana-recognised (ATR) avirulence genes. We describe the identification of 10 amplified fragment length polymorphism (AFLP) markers that define a genetic mapping interval for the ATR1Nd avirulence allele, the presence of which is perceived by the RPP1Nd resistance gene. Furthermore, we have constructed a P. parasitica (At) bacterial artificial chromosome (BAC) library comprising over 630Mb of cloned DNA. We have isolated 16 overlapping clones from the BAC library that form a contig spanning the genetic interval. BAC sequence-derived markers and a total mapping population of 311 F(2) individuals were used to refine the ATR1Nd locus to a 1cM interval that is represented by four BAC clones and spans less than 250kb of DNA. This work demonstrates that map-based cloning techniques are feasible in this organism and provides the critical foundations for cloning ATR1Nd using such a strategy. PMID- 12553935 TI - Deletion of the SNP1 trypsin protease from Stagonospora nodorum reveals another major protease expressed during infection. AB - The wheat fungal pathogen Stagonospora nodorum produces an extracellular trypsin like protease, SNP1, during early stages of hyphal growth on the surface of host leaves and during penetration. Variation of SNP1 mRNA levels and enzyme activity during infection, were correlated with levels of aggressiveness of three wild type isolates. SNP1 was deleted in two wild-type isolates using a gene replacement strategy. SNP1-deleted mutants completely lacked trypsin activity in vitro and on inoculated wheat leaves, but were not reduced in pathogenicity. SNP1 deleted mutants still have 50% of the total alkaline protease activity of wild type. This residual activity comes from a previously undetected alkaline protease with subtilisin-like substrate and inhibitor specificities, which is produced in vitro and on host leaves. We hypothesize that this subtilisin protease may act in concert with SNP1 and may compensate for the loss of trypsin protease activity in the SNP1-deletion mutants. PMID- 12553936 TI - Sequence tag analysis of gene expression during pathogenic growth and microsclerotia development in the vascular wilt pathogen Verticillium dahliae. AB - Two cDNA libraries were constructed from cultures of the vascular wilt fungus Verticillium dahliae, grown either in simulated xylem fluid medium (SXM) or under conditions that induce near-synchronous development of microsclerotia. Expressed sequence tags (ESTs) were obtained for over 1000 clones from each library. Most sequences in the two EST collections were unique; nearly 55% of the translated ESTs had strong similarity to protein sequences in the NCBI nonredundant database. ESTs corresponding to melanin biosynthetic enzymes were exclusive to the developing microsclerotia (DMS) collection, and sequences corresponding to extracellular hydrolases (plant cell wall degrading enzymes) were more abundant in that collection. ESTs corresponding to proteins involved in transport and cell growth were more abundant in the SXM collection. The results of this preliminary analysis suggest that the in vitro growth conditions used here provide useful model systems that will facilitate studies of pathogenesis and microsclerotia development in V. dahliae. PMID- 12553937 TI - Sequence comparison of aflR from different Aspergillus species provides evidence for variability in regulation of aflatoxin production. AB - Aflatoxin contamination of foods and feeds is a world-wide agricultural problem. Aflatoxin production requires expression of the biosynthetic pathway regulatory gene, aflR, which encodes a Cys6Zn2-type DNA-binding protein. Homologs of aflR from Aspergillus nomius, bombycis, parasiticus, flavus, and pseudotamarii were compared to investigate the molecular basis for variation among aflatoxin producing taxa in the regulation of aflatoxin production. Variability was found in putative promoter consensus elements and coding region motifs, including motifs involved in developmental regulation (AbaA, BrlA), regulation of nitrogen source utilization (AreA), and pH regulation (PacC), and in coding region PEST domains. Some of these elements may affect expression of aflJ, a gene divergently transcribed from aflR, that also is required for aflatoxin accumulation. Comparisons of phylogenetic trees obtained with either aligned aflR intergenic region sequence or coding region sequence and the observed divergence in regulatory features among the taxa provide evidence that regulatory signals for aflatoxin production evolved to respond to a variety of environmental stimuli under differential selective pressures. Phylogenetic analyses also suggest that isolates currently assigned to the A. flavus morphotype SBG represent a distinct species and that A. nomius is a diverse paraphyletic assemblage likely to contain several species. PMID- 12553938 TI - Chromosomal heteromorphism and an apparent translocation detected using a BAC contig spanning the mating type locus of Phytophthora infestans. AB - Genetic and physical irregularities associated with the mating type locus of the oomycete, Phytophthora infestans, were revealed by analyzing a contig spanning the locus that was constructed using a bacterial artificial chromosome library. Contigs from both homologs of an A1 strain A/a genotype at mating type locus) had chromosome-specific differences, flanked by regions of similarity. Such heteromorphism was detected within multiple isolates. The mating type locus was narrowed to a 60-70kb interval by genetic mapping of candidate genes, identified using a cDNA library. During these analyses, an unusual isolate of P. infestans was identified in which the mating type determinant had apparently translocated from its location in typical strains. Comparative mapping of the cDNAs between P. infestans and P. parasitica revealed partial synteny between the species however; substantial rearrangements existed and no cDNA was tightly linked to mating type in P. parasitica. These findings add to previous observations of unusual genetic behavior involving mating type in Phytophthora. PMID- 12553939 TI - The interrelationships of actin and hyphal tip growth in the ascomycete Geotrichum candidum. AB - Geotrichum candidum is unusual among reported hyphal ascomycetes in that its hyphae readily stain with phalloidin to reveal actin concentrated in the Spitzenkorper (SPK) and plaques associated with the plasma membrane (PM). Loss of SPK actin, but not the PM plaques, following latrunculin B treatment produces tip swelling, consistent with actin restraining tip morphology or localizing vesicle exocytosis. Tip morphogenesis may also involve a spectrin-like protein which concentrates at the apical PM in plaques unassociated with the actin plaques. Branch formation occurs with growth rates initially about 20% those of leading tips, and does not involve a morphologically detectable SPK, nor SPK-like actin ensembles, indicating the dispensibility of this structure in tip growth. Surprisingly, new tubular tips can form in the continued presence of latrunculin, consistent with alternative cellular systems, such as the spectrin-like protein, substituting for actin's critical functions. PMID- 12553940 TI - Cell wall biogenesis in a double chitin synthase mutant (chsG-/chsE-) of Aspergillus fumigatus. AB - Previous studies (Aufauvre-Brown et al., 1997; Mellado et al., 1996a,b ) have shown that only two genes of the Aspergillus fumigatus chitin synthase family, chsG and chsE, play a role in the morphogenesis of this fungal species. An A. fumigatus strain lacking both chsG (class III CHS) and chsE (class V CHS) genes was constructed by gene replacement of the chsE gene with a copy that has its conserved coding region interrupted by the hph resistance cassette in an A. fumigatus chsG- genetic background. Unexpectedly the double disruption was not lethal. The double mutant AfchsG-/chsE- strain (i) has reduced chitin synthase activity with or without trypsin stimulation, (ii) has a reduced colony radial growth rate, (iii) produces highly branched hyphae, (iv) exhibits aberrant features, such as periodic swellings along the length of the hyphae and a block in conidiation that can be partially restored by an osmotic stabilizer (v) shows alterations in the shape and germination capacity of the conidia, and (vi) has a cell wall that contains half the chitin of the parental strain and is, unexpectedly, highly enriched in alpha-(1-3) glucan. PMID- 12553941 TI - Ras1 controls pheromone expression and response during mating in Cryptococcus neoformans. AB - The Cryptococcus neoformans Ras1 signal transduction pathway controls mating, hyphal differentiation, and the ability of this opportunistic human fungal pathogen to grow at elevated temperatures. To further elucidate how Ras1 signals in this organism, the RAS1 gene was disrupted in the congenic serotype D strain background. Genetic epistasis experiments indicated that Ras1 regulates the mating response through the MAP kinase/pheromone response pathway. In fact, Ras1 is required for the transcriptional induction of elements of the pheromone response pathway. However, the ability of C. neoformans Ras1 to allow growth at 37 degrees C is mediated by a separate signaling pathway. Therefore a single Ras protein may differentially activate distinct downstream targets in response to different signals within the same organism. This conserved signaling motif has been coopted in C. neoformans to regulate mating and morphogenesis in addition to being required for its pathogenic potential. PMID- 12553942 TI - cigA, a light-inducible gene involved in vegetative growth in Mucor circinelloides is regulated by the carotenogenic repressor crgA. AB - A strategy based on the differential-display technique has been applied to identify and to isolate potential target genes controlled by crgA, a negative regulator of the light-inducible carotenogenesis in the fungus Mucor circinelloides. This approach resulted in the identification and cloning of the gene cigA, whose expression is repressed by the crgA gene. After a light pulse, there is an immediate though transient accumulation of transcripts of cigA gene. A similar result was obtained when mycelia were exposed to continuous illumination, suggesting the existence of a photoadaptation mechanism in M. circinelloides. The protein sequence deduced from the nucleotide sequence of cigA shows homology to the mouse and human GDP-fucose protein O-fucosyltransferases. Null cigA mutants were generated by gene replacement. Lack of cigA function does not affect carotenogenesis but provokes a slight reduction in vegetative growth that is independent of light. Introduction of wild-type cigA alleles into a cigA- strain restored the wild-type phenotype for vegetative growth. These results suggest that cigA is involved in vegetative growth and, moreover, that crgA controls cellular processes other than carotenogenesis. PMID- 12553943 TI - The two isoforms of the cAMP-dependent protein kinase catalytic subunit are involved in the control of dimorphism in the human fungal pathogen Candida albicans. AB - We have cloned the Candida albicans TPK2 gene encoding a cAMP-dependent protein kinase (PKA) catalytic subunit and generated a tpk2 homozygous null mutant to assess its ability to germinate in liquid media. N-acetylglucosamine (GlcNAc) induced germ-tube formation was attenuated in the tpk2 strain and enhanced by compounds that are known to increase the PKA activity in situ. Germination was completely blocked in the presence of the myristoylated derivative of the heat stable PKA inhibitor (MyrPKI). These results indicate that TPK1 acts positively in regulating the morphogenetic transition in C. albicans in the absence of the TPK2 gene. We were able to identify an mRNA from this second form of PKA in both wild-type and tpk2 null mutant cells. We found that PKA activity measured in the mutant lacking the TPK2 gene was about 10% of that displayed by the wild-type. The finding that the germinative response of tpk2 null mutant to serum was severely diminished at low serum concentrations indicates that the level of PKA is an important determinant of filamentous growth at low serum concentrations. The extent of germination attained at higher serum concentrations (5%) was similar in the wild-type and in the tpk2 null mutant strains suggesting that under these conditions germination was triggered through a PKA-independent pathway. PMID- 12553944 TI - Insulin revisited. AB - Injury and critical illness are characterised by hyperglycaemia, high free fatty acids and high net protein catabolism, due partly to suppression of insulin secretion in the shock phase and insulin resistance in the flow phase of injury, accompanied by high levels of cytokines and the catabolic hormones cortisol, glucagon and catecholamines. Pre-operative carbohydrate loading reduces post operative insulin resistance and its consequences. Insulin has been shown to reduce the catabolic response as well as controlling hyperglycaemia. In contrast to its sodium retaining properties in normal, obese and diabetic subjects, insulin-glucose-potassium therapy may induce a sodium diuresis in catabolic patients with salt and water overload and in patients with congestive heart failure in whom haemodynamic improvement has also been observed. Diabetic patients with myocardial infarction and cardiac surgery also benefit from insulin treatment. Recent studies have described positive effects on clinical outcome in critical illness. Whether this is due simply to maintenance of euglycaemia or to the other effects of insulin remains to be determined. PMID- 12553945 TI - The liver is an important organ in the metabolism of asymmetrical dimethylarginine (ADMA). AB - BACKGROUND AND AIMS: Asymmetrical dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide (NO) synthase enzymes, whereas symmetrical dimethylarginine (SDMA) competes with arginine transport. Although both dimethylarginines may be important regulators of the arginine-NO pathway, their metabolism is largely unknown. Both dimethylarginines are removed from the body by urinary excretion. However, ADMA is also subject to enzymatic degradation by the enzyme dimethylarginine dimethylaminohydrolase (DDAH), which is highly expressed in the liver. To elucidate the role of the liver in the metabolism of ADMA, we aimed to investigate dimethylarginine handling of the liver in detail. METHODS: Ten male Wistar rats were used for this study. Blood flow was measured using radiolabeled microspheres according to the reference sample method. Concentrations of dimethylarginines were measured by HPLC. The combination of arteriovenous concentration difference and organ blood flow allowed calculation of net organ fluxes and fractional extraction rates. RESULTS: Both the liver (0.89+/-0.11) and the kidney (0.68+/-0.06) showed a high net uptake (nmol/100 g body weight (BW)/min) of ADMA, whereas a significant net uptake of SDMA was only observed in the kidney (0.34+/-0.04). For the liver, fractional extraction rates were 29.5% +/-3.0 for ADMA and 0.0%+/-3.7 for SDMA. Fractional extraction rates of ADMA and SDMA for the kidney were 36.0%+/-2.7 and 31.6%+/-3.8, respectively. CONCLUSIONS: The liver plays an important role in the metabolism of ADMA by taking up large amounts of ADMA from the systemic circulation. PMID- 12553946 TI - Asymmetrical dimethylarginine (ADMA) in critically ill patients: high plasma ADMA concentration is an independent risk factor of ICU mortality. AB - BACKGROUND AND AIMS: Accumulation of asymmetrical dimethylarginine (ADMA) has been linked to endothelial dysfunction, and is an important risk factor for cardiovascular disease. Its elimination from the body is dependent on urinary excretion and degradation by the enzyme dimethylarginine dimethylaminohydrolase. This enzyme is highly expressed in the liver, and in rat studies a high net hepatic uptake of asymmetrical dimethylarginine was found. In critically ill patients, we investigated the relation between indicators of renal and hepatic dysfunction and plasma ADMA concentration, and tested the association between ADMA concentration and outcome. METHODS: We prospectively collected blood samples from a cross-section of critically ill patients (n=52) with clinical evidence of dysfunction of more than two organs. We identified correlates of plasma ADMA concentration with laboratory values, organ failures score and outcome by univariate and multiple regression analyses. RESULTS: In critically ill patients, plasma ADMA concentration was independently related to the presence of hepatic failure (b=0.334, 95% CI: 0.207-0.461; P<0.001), and to lactic acid (b=0.395, 95% CI: 0.230-0.560; P<0.001) and bilirubin (b=0.121, 95% CI: 0.031-0.212; P=0.009) concentration as markers of hepatic function. Twenty-one (40%) patients deceased during their ICU stay. In a logistic regression model, plasma ADMA ranked as the first and strongest predictor for outcome, with a 17-fold (95% CI: 3-100) increased risk for ICU death in patients who were in the highest quartile for ADMA. CONCLUSIONS: In critically ill patients, plasma ADMA concentration is a strong and independent risk factor for ICU mortality, and hepatic dysfunction is the most prominent determinant of ADMA concentration in this population. PMID- 12553947 TI - Effects of 3 days of "postoperative" low caloric feeding with or without bed rest on insulin sensitivity in healthy subjects. AB - BACKGROUND AND AIMS: Insulin resistance after surgery is caused by the surgical trauma and presumably also by other factors, such as starvation and immobilization. The purpose of this study was to evaluate the effect of traditional postsurgical low caloric feeding and bed rest on insulin sensitivity and substrate utilization, in younger and older healthy subjects. METHODS: Twelve healthy subjects underwent hyperinsulinaemic, normoglucaemic clamps and indirect calorimetry before and after 3 days of bed rest and low caloric feeding. Six of the subjects underwent a second study with 3 days of low caloric feeding without bed rest. RESULTS: Insulin sensitivity decreased by 57+/-16 % after low caloric feeding combined with bed rest, with no difference between age groups, and by 56+/-9% after low caloric feeding only. Glucose oxidation decreased, while fat oxidation increased. No significant differences were seen between age groups or between the protocols. CONCLUSIONS: Low caloric feeding, a commonly used nutritional routine in clinical practice, induce marked alterations in insulin sensitivity and substrate utilization. Increasing age or bed rest did not seem to influence this development. These findings suggest that the routine low caloric feeding is capable of contributing to postoperative insulin resistance. PMID- 12553948 TI - The effect of unselected post-operative nutritional supplementation on nutritional status and clinical outcome of orthopaedic patients. AB - BACKGROUND AND AIMS: Nutritional supplements are widely administered in hospitals and can benefit clinical outcome. The aim here was to determine the effect of routine post-operative nutritional supplementation on the nutritional status and clinical outcome of adult orthopaedic patients. METHODS: A prospective controlled study was conducted on two adult orthopaedic wards. Patients in the study group were prescribed two nutritional supplements/day post-operatively. Nutritional and biochemical indices and incidence of clinical complications were observed. RESULTS: Of 181 patients studied, 14 in the supplemented group and 34 in the control (P=0.005) developed major complications. There were 22 occurrences of major complications in the supplemented group and 55 in the control (P=0.0002). There was no significant difference in the number of minor complications between the two groups (P=0.2). There was no statistical difference in changes in nutritional parameters or in albumin or CRP between the two groups. There were significantly greater reductions in transferrin (P=0.002) and in haemoglobin (P=0.002) in the control group at week 1. The median costs of hospital stay were 2068 UK pounds in the supplemented group and 2199 UK pounds in the control. The median cost of additional treatments was 30.16 UK pounds in the supplemented group and 46.23 UK pounds in the control. CONCLUSION: A significant reduction in major complications and in number and costs of additional treatments was seen in the supplemented group. PMID- 12553949 TI - Food production and wastage in relation to nutritional intake in a general district hospital--wastage is not reduced by training the staff. AB - BACKGROUND AND AIMS: To assess the amount of food produced in a hospital kitchen and the amount wasted. To assess the amount of food eaten by patients in relation to their energy needs. To assess whether the food production and wastage could be reduced by training members of the staff. METHODS: The study was carried out in a general district hospital in Denmark. The amount of food produced in the hospital kitchen and returned uneaten (wasted) was determined. In a representative sample of patients, the energy expenditure was calculated and in the same patients, the energy and protein intake was determined. Following training of the hospital staff the first part was repeated. RESULTS: On average, 11.1 MJ and 112 g of protein were ordered per patient per day. From these amounts on average 3.1 MJ and 33 g protein were wasted per patient per day. The total average energy expenditure was calculated to be 7.5 MJ per patient per day; however, on average, the daily energy intake was only 4.5 MJ and daily protein intake 46 g per patient. Sixty percent of the patients showed evidence of malnutrition as judged from the hospital notes. However, the staff only drew attention to this in 20% of the cases. Following training of the hospital staff, a new investigation showed no significant changes in the amount of food ordered and wasted. CONCLUSION: Despite a supply of food, which was much higher than the patients' needs, the patients have only approx. 60% of their energy need covered. We suggest a reorganization of nutrition in hospitals, so that this is made the responsibility of specific staff members. PMID- 12553950 TI - Folate status and a new repletion cut-off value in a group of healthy Majorcan women. AB - BACKGROUND AND AIMS: Folate status is complex, complicated to assess and we lack consensus for laboratory determination. Total plasma homocysteine (Hcy) is a sensitive marker of folate status. The aim of this study was choosing a plasma total homocysteine (tHcy) measurement method and folate repletion level; assess the mutation C677T frequency for methylenetetrahydrofolate reductase (MTHFR) and the prevalence of hyperhomocysteinemia in healthy Majorcan women. METHODS: The measurement methods were compared using 219 women. Folate status assessment was determined by plasma tHcy, serum and erythrocyte folate and C677T for MTHFR in 342 healthy women. The mutation frequency is established with 146 of them. RESULTS: The measurement method comparison is summarized by Y = 1.013 (IC 95% 0.959, 1.069) X +0.829 (IC 95% 0.485, 1.170). The folate repletion level was set at Hcy < or = 8.6 micromol/l. Women (67.1%) are heterozygotic and do not carry the mutation. Homozygotic frequency is 18.5%, significantly higher in women under 40 years (P = 0.033). Hyperhomocysteinemia prevalence is 19.3%, and 51.7% in the younger group. CONCLUSIONS: Hcy determination by polarized fluorescence immunoassay reflects folate status. Levels < or = 8.6 micromol/l suggest folate repletion, which is less frequent in homozygotes. Population frequency for homozygotic-TT alleles for C677T-MTHFR reductase and hyperhomocysteinemia is significantly higher in women under 40 years. PMID- 12553951 TI - Impact of omega-3 fatty acid enriched TPN on leukotriene synthesis by leukocytes after major surgery. AB - Major surgery leads to post-traumatic immune dysregulation which is driven by the activation of potent proinflammatory mediators including the leukotrienes (LTs). The LTs of the four-series derive from arachidonic acid (an omega-6 fatty acid). In contrast, LTs of the five-series are metabolic products of eicosapentaenoic acid (an omega-3 fatty acid) and exert less biological activities. Therapeutical strategies to attenuate proinflammatory signals include the provision of omega-3 fatty acids. Thirty patients with major elective abdominal surgery and an indication for total parenteral nutrition (TPN) were compared in a prospective, double blind, randomized study of two parallel groups. Group 1 (n=14) received an omega-3 fatty acid enriched 20% lipid emulsion (MCT:LCT:fish oil = 5:4:1, MLF541; Lipoplus) for 5 days postoperatively. Group 2 (n=16) received a standard 20% fat emulsion (LCT; Intralipid). The LT release from whole blood leukocytes stimulated with Ca-ionophore was analyzed preoperatively and on postoperative days 1, 6 and 8 by HPLC. There was a significant increase in the generation of LTB(5) (P=0.0035) and in the ratio of LTB(5)/LTB(4) (P=0.0017) the omega-3 group, but not in the reference group after 5 days infusion of the lipid emulsions. The omega-6/omega-3 fatty acid ratio 3:1 of the newly developed MLF541 lipid emulsion is appropriate to increase the synthesis of the biologically less active leukotrienes of the five-series. Nutritive enrichment with omega-3 fatty acids in a balanced ratio with omega-6 fatty acids is an important step to avoid hyperinflammatory situations in patients after major surgery. PMID- 12553952 TI - Carotenoids are decreased in biopsies from colorectal adenomas. AB - A lower intake of carotenoids is associated with an increased risk of colorectal cancer. In order to take advantage of the chemopreventive properties of carotenoids, it is necessary to determine carotenoid concentration at the target tissue. As early stages in the adenoma-carcinoma sequence of colorectal cancer might be susceptible to chemoprevention, we sought to determine carotenoid concentrations in biopsies from colorectal adenomas. METHODS: Biopsies from colorectal adenomas and non-involved mucosa were taken from seven patients. For controls, biopsies were obtained from the ascending and descending colon of patients without polyps (n = 5). Concentration of carotenoids (alpha-, beta carotene, lutein, lycopene, zeaxanthin, beta-cryptoxanthin) were determined by optimizing gradient HPLC-analysis. Results are expressed as pmol/microg DNA. RESULTS: Except for alpha-carotene, all carotenoids could reliably be detected in all specimens. In control patients carotenoid concentrations were highest in the ascending colon, being followed by the descending colon and non-involved mucosa from polyp-carriers. In colorectal adenomas all carotenoids were significantly reduced as compared to-non-involved mucosa (beta-carotene: 0.37 vs 0.19, P<0.03; lycopene: 0.34 vs 0.21, P<0.06, beta-cryptoxanthin: 0.14 vs 0.09, P<0.03, zeaxanthin: 0.18 vs 0.09, P<0.02; lutein: 0.18 vs 0.13,P <0.02). CONCLUSION: All carotenoids investigated are reduced in colorectal adenomas, suggesting that mucosal carotenoids could serve as biomarkers for predisposition to colorectal cancer. Moreover, anti-tumor activity exerted by carotenoids is limited due to mucosal depletion. We speculate that supplementation of a larger array of carotenoids might be beneficial for patients with colorectal adenoma. PMID- 12553953 TI - The effects of fish oil, olive oil, oleic acid and linoleic acid on colorectal neoplastic processes. AB - BACKGROUND AND AIMS: Several nutrients play a significant role in colorectal cancer development, and fats could be among the most determinant. While several studies have shown that the n-3 fatty acids eicosapentaenoic and docosahexaenoic and its main dietary source, fish oil could exert important antineoplastic effects, much less is known about the effects of olive oil and its main fatty acid, oleic acid, and linoleic acid. The aim of these studies is to assess the role of these nutrients in crucial processes involved in colorectal carcinogenesis. METHODS: Caco-2 and HT-29 colorectal cancer cells were supplemented with different fats and their role in apoptosis induction, cell proliferation, and differentiation was studied. COX-2 and Bcl-2 expressions were also assessed. RESULTS: Supplementation with fish oil or olive oil results in an induction of apoptosis and cell differentiation. The latest effect was also induced by oleic and linoleic acid. Fish oil diminishes significantly cell proliferation. Supplementation with fish oil and olive oil results in an early downregulation of COX-2 followed by a decrease in Bcl-2 expression. CONCLUSIONS: Fish oil and olive oil are capable of influencing crucial processes responsible for colorectal cancer development. COX-2 and Bcl-2 may be important mediators of some of these effects. PMID- 12553954 TI - Effects of metabolic control on vitamin E nutritional status in children with type 1 diabetes mellitus. AB - OBJECTIVE: To evaluate the effects of the metabolic control of the insulin dependent diabetes mellitus (IDDM) on the nutritional status of vitamin E. METHODS: A total of 47 children with IDDM and a mean age of 11.91+/-1.60 (mean+/ SD) years were studied, matched for age and sex with 16 healthy children (11.75+/ 1.83 years). Following the American and British Associations for IDDM, we used a classification of "good", "moderate" or "poor" control of the illness: (a) good control [glycosilated haemoglobin (HbAlc) < or =7%]; (b) moderate control (7%< HbAlc < or =8%); and (c) poor control (HbAlc>8%). Serum concentrations of total cholesterol (CHOL), triglycerides (TG), high-density lipoproteins-cholesterol (HDLc), very low- and low-density lipoproteins cholesterol (VLDLc and LDLc), plasma and erythrocyte vitamin E (Vit Ep and Vit Ee) and plasma vitamin A (Vit Ap) were measured in all children. RESULTS: The children with "poor" metabolic control of the illness presented significantly higher plasma concentrations of CHOL, LDLc, VLDLc, LDLc/HDLc, (VLDL+LDL)c/HDLc, TG and Vit Ep; higher indices Vit Ep/Vit Ee and Vit Ep/Vit Ap relative to those with a "good" control of the illness. Direct-linear correlations were found between Vit Ep and the percentage of HbAlc and with the markers of atherogenic risk in the IDDM children. CONCLUSION: There is a significant rise in Vit Ep concentrations as metabolic control of the illness worsens, whereas Vit Ee remained unchanged. These results suggest that vitamin E maintains its nutritional status despite a poor control of the IDDM during childhood. PMID- 12553955 TI - The impact of fluid therapy on nutrient delivery: a prospective evaluation of practice in respiratory intensive care. AB - BACKGROUND AND AIMS: Concurrent therapies are not accounted for nutritional intake. This prospective clinical audit in an intensive care unit aimed at assessing the influence of fluid therapy and drugs containing nutrients on total nutrient delivery. METHODS: Forty-four patients, 25M: 19F, age 63+/-12 (17 83)years, Acute Physiological and Chronic Health Evaluation II: 24+/-9, with length of stay >72h, were evaluated. Basal energy expenditure was estimated by the Harris-Benedict formula. All nutrients conveyed by oral, enteral, parenteral nutrition, IV solutions and drugs were daily registered and summed at the end of respiratory intensive care unit stay. RESULTS: Mean estimated basal energy expenditure (BEE), 1372+/-432 kcal/day, was lower than mean caloric intake 2034+/ 432 kcal/day (P<0.05), >BEE in 54% of patients. When dextrose/propofol were excluded, mean energy intake, 1375+/-430 kcal/day, was similar to BEE; 93% and 86% of patients had an adequate carbohydrate and lipid intake, respectively. Dextrose determined an excessive carbohydrate delivery, higher than 58% of calories, in 43% of patients, while 84% had an excessive lipid delivery due to propofol. Excessive sodium from normal saline was administered to 91% of patients, though 43% had overt oedema. CONCLUSIONS: Hypernutrition ensued from non-quantified fluid therapy/drugs. Sodium loads may aggravate lung and gut dysfunction. The provision of "occult" calories and minerals may be quite substantial and should be taken into consideration. PMID- 12553956 TI - Plasma urea-cycle-related amino acids, ammonium levels, and urinary orotic acid excretion in short-bowel patients managed with an oral diet. AB - BACKGROUND AND AIMS: The small intestine contains several enzymes involved in arginine synthesis and converts glutamine to citrulline, the major compound for endogenous arginine synthesis. This study was conducted to assess the plasma status of urea-cycle intermediates and orotic urinary excretion in short-bowel patients. METHODS: Thirteen stable short-bowel syndrome patients (7 men; 60.2+/ 15.2 years) were studied. Patients were divided into moderately resected (Group A; n=6) and severely resected (Group B; n=7) according to their remnant bowel length (Group A: 61-150 cm; Group B: < or =60 cm). All subjects were consuming an oral diet plus dietetic supplements. Plasma urea-cycle amino acids, ammonium and urinary orotic acid were determined. RESULTS: Plasma glutamine levels were significantly higher in both patient groups than in the control group (P<0.001). Regarding citrulline, Group B levels were significantly lower vs. controls (P<0.001). Comparisons between patient groups showed higher arginine in Group A (P<0.05) and non-statistically lower citrulline in Group B. Blood ammonium and orotic urinary excretion were normal. CONCLUSIONS: Although plasma citrulline and glutamine alterations were found, patients showed no hyperammonemia or orotic aciduria, which suggests a certain degree of adaptation in arginine and related amino acid metabolism, when an adequate dietary supply of arginine is provided. PMID- 12553957 TI - The asymmetrical dimethylarginine (ADMA)-multiple organ failure hypothesis. AB - Development of multiple organ failure is the most complex problem in critically ill patients, and is associated with a high mortality. Asymmetrical dimethylarginine (ADMA) is an endogenously produced inhibitor of nitric oxide synthase, of which the clinical importance is currently being revealed. In Nijveldt et al. (The liver is an important organ in the metabolism of asymmetrical dimethylargenine (ADMA) Clin Nutr 2003; 22: 17-22) we showed that plasma ADMA concentration is elevated in critically ill patients and significantly related to hepatic function. Moreover, plasma ADMA concentration emerged as a strong and independent risk factor for ICU mortality in these patients. Here, we hypothesize that accumulation of ADMA is a causative factor in the development of multiple organ failure by interfering with important physiological functions of nitric oxide production. PMID- 12553964 TI - Discrimination training: a comparison of two procedures for presenting multiple examples within a fading and non-fading paradigm. AB - When teaching discriminations, many researchers and practitioners recommend presenting multiple examples of both the correct and incorrect stimuli. To test this suggestion, we compared two procedures for presenting multiple examples. In one, multiple examples across trials (ME, Across), one correct (S+) and one incorrect (S-) stimulus were presented each trial; examples then changed across trials. In another procedure, multiple examples within trials (ME, Within), three stimuli (either 2 S+'s and 1 S-, or 1 S+ and 2 S-'s) were presented each trial; examples again changed across trials. Two experiments were conducted to test these procedures. The first procedure used a non-fading program to teach discrimination; the second used a fading procedure. In the first experiment, we taught 10 persons to identify words under these two procedures. The former procedure was superior in acquisition; the latter procedure, however, was better under generalization for most participants. In the second experiment, we presented the two procedures within a fading paradigm. The results replicated those in Experiment 1: ME, Across was better for acquisition, but ME, Within was better for generalization. Results were discussed and follow-up studies suggested. PMID- 12553965 TI - The MMPI-168(L) and ADD in assessing psychopathology in individuals with mental retardation: between and within instrument associations. AB - An abbreviated version of the Minnesota Multiphasic Personality Inventory, the MMPI-168(L), modified for use with clients who have moderate or mild mental retardation, was administered to 58 clients, most of whom had co-existing dual psychiatric diagnoses. Another recently developed instrument, the Assessment of Dual Diagnosis (ADD), was administered by interviewing a knowledgeable care giver. Correlations were examined among the raw scores on the 13 ADD scales and T scores of the 13 MMPI-168(L) scales. Contrary to expectations few correlations were found between the scales of the two instruments including scales purported to assess similar psychological constructs. The major exception was the Mania scale of the MMPI-168(L), which correlated moderately well with the Schizophrenia and Dementia scales of the ADD. Client age correlated strongly and negatively with scores on the Conduct Disorder and Sexual Disorder scales of the ADD. Finally, intra-instrument scale correlations were surprisingly large and, from a clinical and diagnostic perspective, meaningful. However, the large number of intra-instrument correlations showed that the scales of both instruments possess considerable overlap, which could make differential diagnosis problematic. It is suggested that it might be necessary to administer both instruments, and carefully consider behavioral history, to accurately diagnose psychiatric disturbances or personality characteristics of individuals with mental retardation. PMID- 12553966 TI - Social impairment in the "Care in the Community" cohort: the effect of deinstitutionalization and changes over time in the community. AB - This paper presents data from a longitudinal collaborative study of The Care in the Community Sample (Cambrdige, Hayes, Knapp, Gould, & Fenyo, 1994; Cambridge et al., 2001; Knapp et al., 1992). The aim of the study was to investigate how social impairment changes are related to the move from institutional to community care using some preliminary analysis of the above data. A measure of social impairment using the Skills and Behavior Interview from the most recent follow-up of this cohort was found to be consistent with Wing's definition of social impairment, when applied to the cohort 12 years after deinsitutionalization. This measure was then used to retrospectively identify social impairment in the same sample (of approximately 250 people) at baseline (in the institution), at 1 year and at 5 years in the community. Prevalence data pointed to high levels of social impairment in the sample (50.1%) in institutions but the decrease to 39.8% after 1 year in the community was not significant (although conversation and social mixing and initiation of conversation and social interaction did improve over time). Once in the community, social impairment in general did not change over time, although there was a significant decrease in conversation and social mixing, non-verbal communication and initiation of conversation and social interaction. These results are compared to other research findings and the implications and limitations of the study discussed. PMID- 12553967 TI - Training phonological awareness skills in children with Down syndrome. AB - Increasingly, children with Down syndrome receive literacy instruction based on a phonological awareness philosophy with the expectation of acquiring functional reading skills. Previous research demonstrates that a phonological awareness based reading programme delivers excellent results in terms of literacy acquisition and improvements in speech production for children with speech and language delays. Unfortunately, little research exists to support the effectiveness of this approach for children with Down syndrome. The current research study examined using a phonological awareness based intervention programme with three children with Down syndrome (aged 7;2, 8;4, and 8;10). A multiple baseline across behaviours design was selected. The intervention programme focused on the key skills of alliteration detection, phoneme isolation, spelling of orthographically regular words and rhyme detection. Two tasks (comprehension of passive structures and spatial structures) were selected as control behaviours. Phoneme segmentation and speech intelligibility were selected to investigate generalisation of intervention targets to other related skill areas. The results indicated that the participants improved the phonological awareness skills targeted in the intervention programme. Unfortunately, no generalisation to other areas of phonological awareness was noted. In summary, the results indicate that children with Down syndrome can benefit from a phonological awareness based approach to literacy. PMID- 12553968 TI - Generalized imitation and the mand: inducing first instances of speech in young children with autism. AB - It has been suggested that the use of a generalized motor imitation sequence before a vocal model may be an effective procedure for teaching nonvocal children with autism to speak. However, the tactic has rarely been empirically demonstrated. The purpose of this experiment was to test the effects of presenting a rapid generalized motor imitation sequence before an opportunity to imitate on the vocal speech of nonvocal children with autism. Participants emitted no vocal imitations during a mand training baseline. During the intervention, a rapid motor imitation sequence was presented before an opportunity to imitate a model's vocalizations. The teacher's presentation of the rapid motor imitation sequence was then faded by presenting an opportunity to vocally imitate without the sequence followed by an opportunity to independently mand. Results of the intervention phase indicated that all of the participants began to vocalize with the generalized motor imitation sequence and that mands were maintained during a follow-up phase and 3-month follow-up probes. PMID- 12553969 TI - A system of assessment for adaptive behavior, social skills, behavioral function, medication side-effects, and psychiatric disorders. AB - A reliable and valid assessment is necessary for the effective delivery of services to those with mental retardation (MR). With constraints placed on financial and human resources, assessment must be comprehensive, yet cost effective. We describe a method of assessment that operates within these constraints using informant-based measures that assess adaptive and maladaptive behavior, psychiatric disorders, behavior function, and medication side-effects. PMID- 12553970 TI - Age- and size-specific patterns of heavy metals in the organs of freshwater fish Abramis brama L. populating a low-contaminated site. AB - Concentrations of cadmium, copper, mercury, lead and zinc were determined by atomic absorption spectrophotometry in the muscle, gill and liver of bream Abramis brama L. to study the relationship between the heavy metal load of fish and their age and size, and the seasonal variation of pollutant loads. Fish were collected from the Western basin of Lake Balaton (Hungary) in October 1999 and May 2000. The average metal concentrations of different organs varied in the following ranges: Cd 0.42-2.10; Cu 1.77-56.2; Hg 0.01-0.19; Pb 0.44-3.24; Zn 10.9 82.5 microg g(-1) dry weight. The highest Cd, Cu, Pb and Zn concentrations were detected in the gill or liver of fish, whereas the highest Hg concentrations were measured in the muscle. In the liver of bream for cadmium, copper and mercury the Pearson correlation analysis revealed positive associations related to age and size (length, net weight), as well as for the mercury load of all three investigated organs. In the muscle and gill the copper, lead and zinc concentrations, similarly to the lead and zinc concentrations of the liver, the associations related to age and size were negative. The correlations between the heavy metal concentrations of organs and the individual condition factors of fish samples proved to have opposite trends compared to those related to the age and size of fish. The seasonal variations in the heavy metal load of bream could be attributed rather to the seasonal change in the condition factor of fish than to variations in the pollutant load of the site. PMID- 12553971 TI - Potential internal loading of phosphorus in a wetland constructed in agricultural land. AB - Wetland construction on agricultural or dairy lands could result in solubilization of phosphorus (P) stored in soils and release to the water column. To study the extent of P flux during the start-up period of a constructed wetland, intact soil-cores from areas used for dairy operations, in Okeechobee, Florida, USA were obtained and flooded with adjacent creek water. In the first 28 day hydraulic-retention period, P concentration in the water column increased several fold due to rapid P flux from impacted soils. A continuous decrease in P flux to the water column until the third hydraulic retention cycle (initial influent P concentration 0.2 mgL(-1)), and constant thereafter suggest that the effect of initial influent P upon long-term P flux from soils could be limited. The initial release maybe due to high concentration of labile P in impacted soils; however, slow dissolution of relatively stable P pools could maintain a steady flux, well above of that observed from non-impacted soils. Water soluble P along with double acid-extractable magnesium explained 76% of the variability in cumulative P flux to the water column. Apparently, co-occurrence of active adsorption-desorption phenomena due to independent maintenance of equilibrium by individual P compounds regulates P dynamics of the water column. The results indicated that equilibrium P concentration of the water column of the wetland would be above 1.3 mgL(-1), which is well above the targeted P level in the water column of the Lake Okeechobee, one of the main water bodies in the area (0.04 mg PL(-1)). This suggests construction of wetlands in agricultural lands could result to substantial internal P loading. However, preventative measures including chemical amendments, establishment of vegetative communities or flushing the initially released P may potentially stabilize the system, and maintain P removal efficiency. PMID- 12553972 TI - New approach to control the methanogenic reactor of a two-phase anaerobic digestion system. AB - A new control strategy for the methanogenic reactor of a two-phase anaerobic digestion system has been developed and successfully tested on the laboratory scale. The control strategy serves the purpose to detect inhibitory effects and to achieve good conversion. The concept is based on the idea that volatile fatty acids (VFA) can be measured in the influent of the methanogenic reactor by means of titration. Thus, information on the output (methane production) and input of the methanogenic reactor is available, and a (carbon) mass balance can be obtained. The control algorithm comprises a proportional/integral structure with the ratio of (a) the methane production rate measured online and (b) a maximum methane production rate expected (derived from the stoichiometry) as a control variable. The manipulated variable is the volumetric feed rate. Results are shown for an experiment with VFA (feed) concentration ramps and for experiments with sodium chloride as inhibitor. PMID- 12553973 TI - Degradation of 2,4,6-trinitrotoluene by immobilized horseradish peroxidase and electrogenerated peroxide. AB - This paper presents horseradish peroxidase (HRP)-catalyzed removal of 2,4,6 trinitrotoluene (TNT) by an electrochemical packed-bed flow reactor operated in a circulating batch mode with the help of in situ generated hydrogen peroxide. HRP immobilized on the reticulated vitreous carbon electrode was prepared for the cyclic voltammetry of 2,4,6-TNT. Effects of pH and temperature on the TNT electroreduction in 0.2M phosphate buffer saturated with oxygen were examined. HRP immobilized carbon electrode was capable of catalyzing the oxidation and detoxification of 44 microM TNT in aqueous solution under optimized conditions. The removal rate of TNT for the electroenzymatic method was much greater than for electrochemical and biochemical methods. Stoichiometric and kinetic studies indicated that the hydrogen peroxide was utilized more effectively in the electroenzymatic method. Denitrification as intermediate reaction was also investigated. PMID- 12553974 TI - Paracetamol oxidation from aqueous solutions by means of ozonation and H2O2/UV system. AB - Paracetamol oxidation from aqueous solutions is studied by means of ozonation and H(2)O(2) photolysis. Both oxidative systems are able to destroy the aromatic ring of the substrate with a partial conversion of the initial carbon content into carbon dioxide. For the adopted experimental conditions mineralization degrees up to 30% and 40% are observed with ozonation and H(2)O(2) photolysis, respectively. Main reaction intermediates and products are identified for both systems by HPLC and GC-MS analyses and a kinetic characterization is achieved. PMID- 12553975 TI - A central fission pathway in alkylphenol ethoxylate biodegradation. AB - A representative alkylphenol Triton X-100 (having 9.5 oxyethylene subunits) was treated over 40 days under the conditions of the continuous flow activated sludge simulation test in a plant with aeration and denitrification chambers. Treated sewage was separated by sequential extraction with ethyl acetate and chloroform. The extracts were analysed by high-performance liquid chromatography-electrospray ionisation-mass spectrometry (HPLC-ESI-MS). Single ion chromatograms of the chloroform extracts showed the presence of neutral, mono- and di-carboxylated poly(ethylene glycols). This is evidence of the central fission of alkylphenol ethoxylates (APE). Simultaneously, the APE having omega-carboxylated oxyethylene chains were identified. This is the evidence that apart from central fission, the omega-oxidation oxyethylene chain pathway also occurs. PMID- 12553976 TI - Heterogeneity of natural organic matter from the Chena River, Alaska. AB - Water samples were collected in July 2001 from the Chena River in central Alaska. The natural organic matter (NOM) was size fractionated into particulate (POM,>0.45 microm), colloidal (COM,1kDa-0.45 microm) and dissolved (DOM,<1k Da) organic matter fractions, using filtration and ultrafiltration. The size fractionated organic matter was then analyzed for organic carbon (OC) and nitrogen (N), isotopic (delta13C and delta15N) and molecular composition, using continuous flow isotope ratio mass spectrometry and pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS). Results of phase partitioning showed that, on average, about 6% of OC and 16% of N occurred in the form of POM while 66% of OC and 57% of N occurred in the form of COM, and 28% of the OC and 27% of the N were in the DOM form. Organic matter in the river water was found to be highly heterogeneous in terms of chemical composition and isotopic signatures. The C/N ratio was as low as 16+/-1 in the POM (n=2) to as high as 48+/-1 in the COM (n=3) and 38+/-4 in the DOM (n=3), suggesting a diagenetically younger POM. Values of delta13C increased with decreasing size, varying from -29.59+/-0.45% in the POM to -27.47+/-0.06% in the COM to -16.93+/-0.08% in the DOM. In contrast, values of delta15N decreased with decreasing size, from 2.64% in POM to 1.64% in COM to 1.33% in DOM. These results, together with radiocarbon measurements, suggest a preferential decomposition of lighter C isotope (12C) and heavier N isotopic (15N) from POM to COM to DOM. Results of py-GC/MS showed that the percentage of polysaccharides decreased with decreasing size, further supporting a degradation pathway of NOM from POM to COM and DOM in Chena River waters. More studies are needed to examine the seasonal and spatial variations of size fractionated organic matter. PMID- 12553977 TI - Ozonation of polycyclic aromatic hydrocarbons in oil/water-emulsions: mass transfer and reaction kinetics. AB - The ozonation of highly condensed polycyclic aromatic hydrocarbons (PAH) was studied in oil/water-emulsions, which are comparable to poorly water-soluble PAH in industrial wastewaters and at contaminated sites. As there was a lack of knowledge about the ozonation in oil/water-emulsions, first the ozone mass transfer was studied and optimized from the gas to the water phase and from the water to the oil phase. The ratio of mass transfer and oxidation reaction was determined by the Hatta-number and revealed a slow, quasi homogeneous reaction of ozone with PAH inside the oil droplets. Because the ozone gas concentration had no influence under the optimized conditions, the selective PAH-ozonation could be described microkinetically by a direct ozone reaction of pseudo-first order regarding PAH-concentrations. The determined PAH mean reaction rate constants of 1.02 min(-1) in oil/water-emulsions are in the upper range as found for PAH dissolved in water. These results give a new insight into the ozonation in the three-phase systems and into the treatment of highly condensed, hardly biodegradable PAH. PMID- 12553978 TI - Effect of specific gas loading rate on thermophilic (55 degrees C) acidifying (pH 6) and sulfate reducing granular sludge reactors. AB - The effect of the specific gas loading rate on the acidifying, sulfate reducing and sulfur removal capacity of thermophilic (55 degrees C; pH 6.0) granular sludge bed reactors treating partly acidified wastewater was investigated. A comparison was made between a regular UASB reactor and a UASB reactor continuously sparged with N(2) at a specific gas loading rate of 30 m(3)m(-2)d( 1). Both UASB reactors (upflow velocity 1.0 mh(-1), hydraulic retention time about 5h) were fed a synthetic wastewater containing starch, sucrose, lactate, propionate and acetate and a low sulfate concentration (COD/SO(4)(2-) ratio of 10) at volumetric organic loading rates (OLR) ranging from 4.0 to 49.8 gCODl(-1) reactord(-1). Immediately after imposing an OLR of 25 gCODl(-1) reactord(-1), the acidification and sulfate reduction efficiency dropped to 80% and 30%, respectively, in the UASB reactor. Both efficiencies recovered slowly to 100% during the course of the experiment. In the N(2) sparged reactor, both the acidification and sulfate reduction efficiency remained 100% following the OLR increase to 25 gCODl(-1) reactord(-1). However, the sulfate reduction efficiency gradually decreased to about 20% at the end of the experiment. The biogas (CO(2) and CH(4)) production rate in the UASB was very low, i.e. <3l biogasl(-1) reactorday(-1), resulting in negligible amounts (<20%) of H(2)S stripped from the reactor liquid. The total H(2)S concentration of the N(2) sparged UASB reactor effluent was always below 25 mgl(-1), but incomplete sulfate reduction kept the maximal H(2)S stripping efficiency below 70%. PMID- 12553979 TI - Harmful effects of un-ionised ammonia on the zooplankton community in a deep waste treatment pond. AB - The harmful effects of NH(3) on the zooplankton community in a deep waste treatment pond were evaluated under natural conditions. The pond, supplied with secondary effluent from a conventional urban wastewater treatment plant, was designed to improve water quality for agricultural reuse.The aim of the study was to verify the hypothesis suggested in Arauzo et al. (Water Environ. Res. 34(14), 3666) that during phytoplankton blooms in the stratification periods high un ionised ammonia content values, due to an intense photosynthetic activity and high related pH, lead to a decrease in zooplankton biomass and, thus, to a collapse of the treatment process efficiency. Empirical models were developed to determine relationships between phytoplankton biomass, pH and NH(3) levels. They provided an easy and quick method of detecting when the system was liable to collapse due to the NH(3) effect on the zooplankton community and offered the possibility of adopting measures to guarantee water quality at the effluent. A significant decrease in zooplankton community biomass was observed at un-ionised ammonia levels over 2.5 mgL(-1). PMID- 12553980 TI - Pesticides in Portuguese surface and ground waters. AB - Pesticides used in Portuguese agricultural areas have been found in surface and ground waters. In the surface water collected in three river basins from 1983 to 1999, insecticides and herbicides were detected from the monitored pesticides, particularly atrazine, chlorfenvinphos (Z+E), alpha- and beta-endosulfan, lindane, molinate and simazine, reaching the maximum values, respectively, of 0.63, 31.6, 0.18 microg/L (alpha-endosulfan), 0.18 microg/L (beta-endosulfan), 0.24, 48 and 0.3 microg/L. In the ground water collected from the wells of seven agricultural areas from 1991 to 1998, several monitored herbicides were detected: alachlor, atrazine, metolachlor, metribuzine and simazine, reaching the maximum concentration values of 13, 30, 56, 1.4 and 0.4 microg/L, respectively. The herbicides more frequently detected were atrazine (64%), simazine (45%) and alachlor (25%). Other than these, the monitored pesticides can be present in Portuguese surface and ground waters. Therefore, to improve the analytical conditions, the use of multiresidue methods and automated techniques are desirable in future work. PMID- 12553981 TI - Impact of UV disinfection on microbially available phosphorus, organic carbon, and microbial growth in drinking water. AB - UV irradiation at a wavelength of 253.7 nm (UV(254)) is commonly used for drinking water disinfection. UV radiation is known to convert organically combined phosphorus to orthophosphate and to degrade natural organic matter. We studied if UV disinfection increases the amount of microbially available forms of organic carbon and phosphorus in drinking waters with different characteristics, and if these changes in water chemical quality could enhance the microbial growth in drinking water. The UV(254) dose (15-50 mWs/cm(2)) used in waterworks reduced the concentration of assimilable organic carbon and the sum of the molecular size fractions. The release of microbially available phosphorus needed higher doses (204 mWs/cm(2)) of UV(254) radiation. Of bacteria in drinking water, 90% were inactivated with UV(254)-irradiation doses below 50 mWs/cm(2). A high dose (501 mWs/cm(2)) of UV(254) radiation inhibited the microbial growth in water. PMID- 12553982 TI - Sedimentation behaviour of sludge particles in a biogas tower reactor and the function of a hydrostatically pressurized sedimenter. AB - It was found that anaerobic sludge particles contain gas bubbles. Due to the compressibility of the bubbles, which are entrapped in the sludge agglomerates, a pressure-dependent sedimentation characteristic of the sludge particles was found and mathematically described. Enhanced hydrostatic pressure results in a considerable increase of the settling velocity of sludge particles. On this basis the concept of a hydrostatically pressurized sedimenter is presented and its performance analyzed. Due to the hydrostatical pressurization of the sedimenter the produced gas is kept in the liquid phase until the liquid phase is saturated. To avoid a degassing of the liquid which results in the undesired release of gas bubbles a maximum liquid dwell time in the sedimenter must not be exceeded. PMID- 12553983 TI - Some effects of aqueous silica on the corrosion of iron. AB - Silica is an important natural component of ground and surface waters, and is sometimes added as an inhibitor to control "red water" problems caused by corroding iron pipes. However, the effect of silicates on many aspects of iron corrosion has never been assessed. Experiments with water containing 0.5, 10, 25 or 50mg/L of SiO(2) demonstrated a significant interplay between aqueous silica and iron corrosion. During this 4-month experiment, higher levels of silica caused more iron release to the water and decreased the size of suspended iron particles. The process of iron corrosion also changed aqueous silica concentrations; silica was released into the water from the cast iron during corrosion and was removed from the water by incorporation into the scale layer. Silica also affected the type of scale that formed on the iron coupons. Scale at the lower silica concentrations was fairly uniform and easy to remove from the coupons, while the scale from the high silica reactors was more dense, and was more difficult to remove. Scale from the high concentration silica reactor also developed tall tubercles, and hydrogen gas-containing bubbles were channeled to solution through these tubercles. Iron corrosion occurring via the evolution was significant under all experimental conditions. PMID- 12553984 TI - Recovery of ammonium sulfate from fermentation waste by electrodialysis. AB - Electrodialysis experiments of the lysine fermentation waste were performed to generate demineralized feed and ammonium sulfate, which can be utilized as a fertilizer and an animal feed, respectively. The electrodialysis performances were compared for different ion exchange membranes in terms of ammonium sulfate removal rate, resistance and conductivity change. Analysis of fouling phenomena revealed that organics fouled ion exchange membranes reversibly in electrodialysis of the fermentation waste. In this study, mitigation of membrane fouling with the pulsed electric field was examined for the electrodialysis of the fermentation waste containing strong foulants. The half-wave power reduced membrane fouling significantly. For a quantitative measure of the membrane fouling tendency, a membrane fouling index for electrodialysis was used. This study showed the potential use of pulsed power as an effective fouling mitigation method for the electrodialysis of fermentation waste. PMID- 12553985 TI - The role of inorganic carbon limitation in biological nitrogen removal of extremely ammonia concentrated wastewater. AB - It is clear from the fundamental biochemical processes that nitrification of extremely concentrated ammonia loads requires-among others-(1) sufficient alkalinity to buffer acidification and (2) bicarbonate as the substrate for the autotrophic biomass. However, at low pH values the aeration process causes CO(2) stripping and consequently a decrease of the available inorganic carbon. In order to analyse such complex interactions, we suggest in this paper an enhanced version of the widely acknowledged IWA (formerly IAWQ) activated sludge models. These model enlargements comprise an ion-balance for the calculation of the pH value and of dissociation species, a balance of inorganic carbon and a more detailed description of the relevant N-elimination processes and their inhibitions. The model was successfully employed to optimise a treatment strategy for rejection-water and landfill leachate (500-2000 mg ammonia-Nl(-1), COD/N ratio of 0.25-4). Detailed data from two full-scale rejection-water treatment plants were used for systems identification, model calibration and validation. The results suggest that inhibition and limitation by nitrous acid (HNO(2)) and unionised ammonia (NH(3)) have often been overestimated. In this investigation the bicarbonate concentration proved to be crucial for the process. The optimisation of the bicarbonate concentration in the reactor could improve the nitrozation rate up to 100mg NH(4)(+)-Nl(-1)h(-1). PMID- 12553986 TI - Removal of Giardia and Cryptosporidium in drinking water treatment: a pilot-scale study. AB - Giardia and Cryptosporidium have emerged as waterborne pathogens of concern for public health. The aim of this study is to examine both parasites in the water samples taken from three pilot-scale plant processes located in southern Taiwan, to upgrade the current facilities. Three processes include: conventional process without prechlorination (Process 1), conventional process plus ozonation and pellet softening (Process 2), and integrated membrane process (MF plus NF) followed conventional process (Process 3). The detection methods of both parasites are modified from USEPA Methods 1622 and 1623. Results indicated that coagulation, sedimentation and filtration removed the most percentage of both protozoan parasites. The pre-ozonation step can destruct both parasites, especially for Giardia cysts. The microfiltration systems can intercept Giardia cysts and Cryptosporidium oocysts completely. A significant correlation between water turbidity and Cryptosporidium oocysts was found in this study. The similar results were also found between three kinds of particles (phi=3-5,5-8 and 8-10 microm) and Cryptosporidium oocysts. PMID- 12553987 TI - Enhanced settleability and dewaterability of fungal treated domestic wastewater sludge by liquid state bioconversion process. AB - A study was conducted to evaluate the settleability and dewaterability of fungal treated and untreated sludge using liquid state bioconversion process. The fungal mixed culture of Aspergillus niger and Penicillium corylophilum was used for fungal pretreatment of wastewater sludge. The fungal strains immobilized/entrapped on sludge particles with the formation of pellets and enhanced the separation process. The results presented in this study showed that the sludge particles (pellets) size of 2-5mm of diameter were formed with the microbial treatment of sludge after 2 days of fermentation that contained maximum 33.7% of total particles with 3-3.5mm of diameter. The settling rate (measured as total suspended solids (TSS) concentration, 130 mg/l) was faster in treated sludge than untreated sludge (TSS concentration, 440 mg/l) after 1 min of settling time. In 1 min of settling operation, 86.45% of TSS was settled in treated sludge while 4.35% of TSS settled in raw sludge. Lower turbidity was observed in treated sludge as compared to untreated sludge. The results to specific resistance to filtration (SRF) revealed that the fungal inoculum had significant potentiality to reduce SRF by 99.8% and 98.7% for 1% w/w and 4% w/w of TSS sludge, respectively. The optimum fermentation period recorded was 3 days for 1% w/w sludge and 6 days for 4% w/w sludge, respectively, for dewaterability test. PMID- 12553988 TI - Solar photocatalytic thin film cascade reactor for treatment of benzoic acid containing wastewater. AB - A solar photocatalytic cascade reactor was constructed to study the photocatalytic oxidation of benzoic acid in water under various experimental and weather conditions at HKUST. Nine stainless steel plates coated with TiO(2) catalyst were arranged in a cascade configuration in the reactor. Photolytic degradation and adsorption were confirmed to be insignificant total organic carbon (TOC) removal mechanisms. A turbulent flow pattern and, hence, improved mixing in the liquid film were achieved due to the unique cascade design of the reactor. The photoinduced consumption of oxygen during reactions was demonstrated in a sample experiment. The proposed rate equations provided good fits to 90 data points from 17 experiments. The regression results showed that the TOC removal rates averaged over 30 min intervals did not illustrate significant dependence on TOC(0) and that I(mean) was more important in affecting the photocatalytic process within the ranges of the data examined. The percentage removal of TOC in 7 l of 100 mg/l (or 100 ppm) benzoic acid solutions increased from 30% to 83% by adding 10 ml of hydrogen peroxide solution (30 wt%). Hydrogen peroxide was also shown to enhance the efficiency of the degradation process at elevated temperatures. Ortho-, meta- and para-hydroxybenzoic acids were identified by HPLC analysis as the intermediates of benzoic acid during reactions without the addition of hydrogen peroxide solutions. PMID- 12553989 TI - Seasonal change and fate of coliphages infected to Escherichia coli O157:H7 in a wastewater treatment plant. AB - Seasonal change of virulent phage infected to two E. coli O157:H7 strains (O:157 phage) in the influent of a domestic wastewater treatment plant in the central part of Japan and fate of O:157-phage in the plant were monitored almost monthly from March 2001 to February 2002. Coliphage infected to nonpathogenic E. coli O157:H7 ATCC43888 (43888-phage) was detected for 1 year. On the other hand, phage infected to pathogenic E. coli O157:H7 EDL933 (EDL-phage) was detected intermittently. Concentration of EDL-phage was almost one-tenth of that of 43888 phage. The progressive decrease in phage concentration with the treatment steps was observed. No phage was detected in the supernatant from the secondary settling tank and effluent. PCR amplification of the Stx 2 gene that encodes Shiga toxin (Stx) was observed when O:157-phage concentration in the influent was high x10(3) PFU/ml order. Concentration and percentage of suspended O:157-phage decreased with the progress of the wastewater treatment. 933W phage, which encodes Stx 2 gene, was more fragile and sensitive to chlorination than T4 phage. However, addition of 0.02 mg/l chlorine, in conformance with the required concentration of the plant, did not affect the viability of T4 and 933 W phages. On the other hand, 1mg/l chlorine inactivated the 933 W phage significantly. PMID- 12553990 TI - Ground water pollution by roof runoff infiltration evidenced with multi-tracer experiments. AB - The infiltration of urban roof runoff into well permeable subsurface material may have adverse effects on the ground water quality and endanger drinking water resources. Precipitation water from three different roofs of an industrial complex was channelled to a pit and infiltrated into a perialpine glaciofluvial gravel-and-sand aquifer. A shaft was constructed at the bottom of the pit and equipped with an array of TDR probes, lysimeters and suction cups that allowed measuring and sampling soil water at different depths. A fast infiltration flow was observed during natural rainfall events and during artificial infiltration experiments. For a better understanding of the behaviour of contaminants, experiments were conducted with cocktails of compounds of different reactivity (ammonium, strontium, atratone) and of non-reactive tracers (uranine, bromide, naphthionate), which represent different classes of pollutants. The experiment identified cation exchange reactions influencing the composition of the infiltrating water. These processes occurred under preferential flow conditions in macropores of the material. Measuring concentration changes under the controlled inflow of tracer experiments, the pollution potential was found to be high. Non-reactive tracers exhibited fast breakthrough and little sorption. PMID- 12553991 TI - Catalytic wet peroxide oxidation of phenol over Fe-exchanged pillared beidellite. AB - This study presents an evaluation of the catalytic performances of a Fe-exchanged Al-pillared synthetic beidellite for the wet hydrogen peroxide oxidation of phenolic aqueous wastes. The catalyst was prepared by a cation doping technique, its properties being determined by DRX, BET and chemical analysis techniques. All the tests were performed on a laboratory scale set-up. Important factors affecting catalyst activity and phenol removal efficiencies were studied, i.e. the effect of pH, temperature, catalyst concentration and the stability of the catalyst. The experimental results indicate that the use of this catalyst allows a total elimination of phenol and a significant removal of chemical oxygen demand, without significant leaching of Fe ions. Thus, considering the lowest Fe concentrations in solution after oxidation, at pH=5, 50 degrees C, and 180 min. COD removal efficiency of 87.9% was obtained. It was also observed that by using this catalyst, it is possible to extend the range of pH values for which Fenton type oxidations can occur. PMID- 12553992 TI - Formation of aldehydes upon ClO2 disinfection. AB - Carbonyl compounds are considered to be the most common ozonation by-products. Apart from well-known chlorites and chlorates, chlorine dioxide (ClO(2)) also generates organic by-products. The goal of this paper is an investigation into the influence of disinfection with ClO(2) on the aldehydes formation. Three types of waters were disinfected with different doses of ClO(2) at the various pH values. The results of size exclusion chromatography show that the reaction of ClO(2) with humic fraction of natural organic matter proceeds mainly on an aromatic part of the molecules. The results obtained indicate that the level of carbonyls concentration can significantly increase with the time of ClO(2) reaction with carbonyl precursors in treated water. There is no noticeable correlation between quantity of aldehydes and pH value of disinfected water. The range of "productivity" of aldehydes in water treated with chlorine dioxide is similar to the range estimated for the carbonyls formed upon ozonation. PMID- 12553993 TI - Pretreatment of Afyon alcaloide factory's wastewater by wet air oxidation (WAO). AB - In this study, pretreatment of Afyon (Turkey) alcaloide factory wastewater, a typical high strength industrial wastewater (chemical oxygen demand (COD)=26.65 kgm(-3), biological oxygen demand (BOD(5))=3.95 kgm(-3)), was carried out by wet air oxidation process. The process was performed in a 0.75 litre specially designed bubble reactor. Experiments were conducted to see the advantages of one stage and two-stage oxidation and the effects of pressure, pH, temperature, catalyst type, catalyst loading and air or oxygen as gas source on the oxidation of the wastewater. In addition, BOD(5)/COD ratios of the effluents, which are generally regarded as an important index of biodegradability of a high-strength industrial wastewater, were determined at the end of some runs. After a 2h oxidation (T=150 degrees C, P=0.65 MPa, airflowrate=1.57 x 10(-5)m(3)s(-1), pH=7.0), the BOD(5)/COD ratio was increased from 0.15 to above 0.5 by using the salts of metals such as Co(2+),Fe(2+),Fe(2+)+Ni(2+),Cu(2+)+Mn(2+) as catalyst. PMID- 12553994 TI - Pre-treatment optimisation studies for secondary effluent reclamation with reverse osmosis. AB - Physical-chemical pre-treatment was used for the reverse osmosis unit for reclamation of secondary effluents. The pilot plant was equipped with a variety of tertiary treatment units to prevent fouling and biofouling of the cellulose acetate reverse osmosis membranes used. The optimisation of pre-treatment involved application of various concentrations of lime to raise the pH to 10.3 12.1, and to stabilise the sludge generated, as well as different dosages of ferric chloride (15, 20, and 25 mg/L) for the coagulation and solid-liquid separation. Sodium hypochlorite (8 mg/L) and UV disinfection are used for microbiological control. The water quality obtained, under the optimum conditions (pH=10.5; FeCl(3): 25 mg/L; anionic flocculant: 0.5 mg/L; sodium hypochlorite: 8 mg/L) was high, showing an average conductivity of 66 microS/cm and low COD values 4 mg O(2)/L. The product water is suitable for injection into a groundwater aquifer to counteract seawater intrusion. PMID- 12553995 TI - Ozonation of naphthalene sulfonic acids in aqueous solutions: part II- relationships of their COD, TOC removal and the frontier orbital energies. AB - This paper evaluated the COD, TOC removal and molecular frontier energies in the ozonation of naphthalene sulfonic acids (NSAs). It was found that both COD and TOC degradation of the 11 compounds followed the pseudo-first-order kinetic and the reaction rate constants with ozone varied widely. A linear relationship between logarithm of global COD degradation rate constant and the energy of the highest occupied molecular orbit, E(homo), was obtained for these NSAs with a regression coefficient of 0.88 and a slope of 0.4672 eV(-1). TOC degradation kinetics also reached similar results with a regression coefficient of 0.90 and a slope of 0.9336 eV(-1). The results of correlation analysis suggested that TOC and COD removal in ozonation of NSAs follow the frontier orbital theory and can be predicted by E(homo), which indicated that electrophilic effect was the main factor in the ozonation of NSAs. Therefore, it is possible to improve the biodegradability of dyestuff wastewater bearing mainly NSAs by ozonation at an accepted cost and evaluate the economy of ozonation process. PMID- 12553996 TI - Characteristics of microfiltration membranes in a membrane coupled sequencing batch reactor system. AB - Factors affecting filtration performance were investigated in a sequencing batch reactor (SBR) coupled with a submerged microfiltration module. Special bioreactors for aerobic and anoxic phases were specifically designed in order to differentiate the effect of dissolved oxygen (DO) from that of mixing intensity, on membrane filterability. When the filterability of a submerged microfilter was examined at each SBR phase, DO concentration, as well as mixing intensity proved to have a major influence on the membrane performance regardless of the SBR phase. A higher DO concentration resulted in a slower rise in TMP, corresponding to less membrane fouling, which was investigated in depth through a series of analyses including resistance measurements and compressibility of the cake layer as well as particle sizes as a functions of DO for both aerobic and anoxic phases in SBR. PMID- 12553998 TI - The management of mentally impaired offenders within the South Australian criminal justice system. PMID- 12553999 TI - Seattle's mental health courts: early indicators of effectiveness. PMID- 12554001 TI - The South African court for sexual offences. PMID- 12554000 TI - The Broward Mental Health Court: process, outcomes, and service utilization. PMID- 12554002 TI - Treating adolescent substance abuse problems in the juvenile drug court. PMID- 12554003 TI - Are internists and cardiologists two of the same kind? PMID- 12554004 TI - Developments in internal medicine. PMID- 12554005 TI - Clinical and epidemiological aspects of cardiomyopathies: a critical review of current knowledge. AB - Five years after the second report of the WHO/ISFC Task Force on the definition and classification of cardiomyopathies (CM), a critical review of the matter appears well-timed. The need for a correct definition of myocardial diseases is emphasized by considering them the result of a 'direct' injury due to different known and unknown causes and not a consequence of ischemic disease or of pressure and/or volume overload. This is in order to eliminate terms like ischemic CM, valvular CM, and hypertensive CM, which are a source of confusion. The concept of myocardial injury is also reviewed. This should not only include the structural/organic macroscopic injury, but also the subcellular, ultrastructural, and molecular damage (mostly of genetic origin) of the contracting element proteins, of citosol, sarcolemma and cell membrane ion channels. As the myocardium is a complex structure, made of common fibers and of specific conduction tissue, injury may be clinically identified either by ventricular function impairment or by bioelectric function defects, i.e. tachyarrhythmias and/or bradyarrhythmias, which sometimes are the unique manifestation of the disease (arrhythmogenic CM, in the strict sense). On the basis of the morpho functional alterations, CMs may be classified as dilated CM (which could be better identified as hypokinetic CM, referring to the functional aspect, because the morphologic aspect is not always present), hypertrophic CM, restrictive CM, and arrhythmogenic CM (including not only arrhythmogenic right ventricular CM, but also other forms, like the so-called arrhythmias of the 'apparently' healthy heart, due to 'occult' myocardial injury). Moreover, these forms may present in association, like mixed CM (dilated-arrhythmogenic, dilated-hypertrophic, etc.). From an etiologic point of view, it is advisable to maintain the distinction between specific CM, due to a known cause, and primary or idiopathic CM, including, together with sporadic forms of an unknown origin, familial forms of a genetic origin, depending on alterations of contractile or regulating functional proteins, when myocardial injury is the sole manifestation (idiopathic) of clinical picture. The most modern etiopathogenetic, pathophysiological, and clinical features of each form of CM are briefly described in order to suggest a complete definition of the disease and to state a clinical-epidemiological setting that encompasses the current knowledge. PMID- 12554006 TI - Oral chemotherapy in colorectal cancer treatment: review of the literature. AB - Recent advances in anticancer treatment have focused on the development of oral anticancer agents with the intention of improving the patients' quality of life as well as providing therapeutic alternatives to intravenous chemotherapy. Until agents such as oxaliplatin and irinotecan became available, the treatment of colorectal cancer, one the most common cancers diagnosed in industralized countries, was mainly based on 5-fluorouracil modulation. The overwhelming majority of these new drugs are pyrimidine analogues intended to replace intravenous treatment or to make the therapy more acceptable to the patients. In this article, the use of oral chemotherapy, alone or in combination with radiotherapy, in colorectal cancer is reviewed and updated. The rationale for using oral compounds is discussed and newer agents, such as oral camptothecin analogues and antiangiogenic agents, are presented with the results of their clinical and preclinical developments. PMID- 12554007 TI - Is clinical diagnosis of heart failure reliable?-Clinical judgement of cardiologists versus internists. AB - BACKGROUND: Heart failure is a major cause of morbidity and mortality. Its diagnosis is mainly clinical. Most patients are seen by cardiologists and internists. However, it is not known whether clinical practices vary by specialty. We sought to evaluate the concordance in the clinical judgement of heart failure between cardiologists and internists. METHODS: Clinical judgement was investigated in 17 cardiologists and 25 internists from four district hospitals using the probability assessment of heart failure diagnosis in 30 case histories based on real patients. Use of clinical information in the diagnostic assessment was defined by the regression coefficients of multiple regression analysis, in which the diagnostic probability was the dependent variable and the clinical criteria the independent variables. The importance attributed to clinical variables, as reported by doctors in a questionnaire, and that of clinical practice, as expressed by the magnitude of the regression coefficients, were compared. RESULTS: We found no significant difference between cardiologists and internists. However, within each group of specialists, there was a wide inter observer variation in the probability assessment of heart failure in the same case histories. The probability ranged from 25.6 to 83%. The relative importance of clinical variables actually used in diagnostic assessment was different from that reported by doctors. CONCLUSIONS: Cardiologists and internists do not differ in their clinical judgement of heart failure. However, within each group there can be wide discrepancies in the evaluation of the same case histories. This may be related to the different use of clinical information, as indicated by the wide confidence intervals of regression coefficients for clinical criteria. The way doctors use clinical information in practice differs from how they think they use it. PMID- 12554009 TI - Low prevalence of HCV, HIV, and HTLV-I/II infection markers in northwestern Greece: results of a 3-year prospective donor study (1995-1997). AB - Background: The risk of infection with transfusion-transmitted viruses has been reduced remarkably. A zero-risk blood supply, however, remains a popular goal. A 3-year prospective donor study was conducted in the Epirus region of Greece to determine the prevalence of human immunodeficiency virus (HIV), human T lymphotropic virus (HTLV), hepatitis B virus, and hepatitis C virus (HCV). Herein, we report the prevalence of HIV, HTLV, and HCV infection markers in this area. Methodology: Between January 1, 1995 and December 31, 1997, 6696 donors were investigated for the presence of anti-HIV, anti-HTLV, and anti-HCV antibodies using standard enzyme immunoassays (EIA). Every sample with anti-HCV reactivity by third-generation EIA was further investigated using a third generation recombinant immunoblot assay (RIBA 3.0) and HCV-RNA by a combination of polymerase chain reaction (PCR) and DNA EIA. Results: None of the donors tested positive for anti-HIV or anti-HTLV antibodies. In contrast, anti-HCV was detected in 41 donors (0.61%). Using a RIBA 3.0 test, eight donors tested positive and eight had indeterminate results, while 25 tested negative. Seven of the eight donors with both EIA and RIBA 3.0 reactivity had increased levels of aminotransferases and detectable serum HCV-RNA. The remaining 34 donors had repeatedly normal aminotransferases and three times negative HCV-RNA. Liver biopsy was performed in anti-HCV/HCV-RNA-positive donors (7/41). The lesions were compatible with chronic hepatitis C in all of them. Conclusion: A zero prevalence of HIV and HTLV infection markers was found. Although the number of annual donations in this study was relatively low, the negative data for HIV and HTLV clearly indicate that rates of these infections are low in our region and that infected donors will be seen infrequently. HCV infection in blood donors remains very low in our region and is similar to the data reported in other industrialized countries. In fact, the prevalence of definite HCV infection seems to be very low (7/6696; 0.1%). However, a significant proportion of anti-HCV reactive donors by third-generation EIA (33/41) had indeterminate or negative results by the RIBA 3.0. The latter donors were repeatedly negative for HCV-RNA. This finding may indicate that some donors tested false-positive for anti-HCV, although the possibility of true HCV infection contracted in the distant past cannot be excluded. In our opinion, close attention to mandatory principles of transfusion medicine, along with the screening of plasma donors using nucleic acid amplification technology, are the only methods that can further ensure the safety of our blood supply. PMID- 12554008 TI - Serum levels of insulin-like growth factor (IGF)-I and IGF binding protein (IGFBP)-1 to -6 and their relationship to bone metabolism in osteoporosis patients. AB - BACKGROUND: Insulin-like growth factor (IGF) system components are important regulators of bone formation. Alterations of individual IGF system components have been described in osteoporosis (OP) patients; however, no study has addressed changes in free IGF-I and in all six IGF binding proteins (IGFBPs). METHODS: A cross-sectional study was performed in 45 OP patients and 100 healthy matched controls. Serum levels of free and total insulin-like growth factor I (IGF-I), IGFBP-1 through -6, intact parathyroid hormone (PTH), 25-OH-vitamin D(3) (25OHD(3)), 1,25-(OH)(2)-vitamin D(3) (1,25-(OH)(2)D(3)), osteocalcin (OSC), bone alkaline phosphatase (B-ALP), and carboxyterminal propeptide of type-I procollagen (PICP) were measured with specific assays. Bone mineral density (BMD) of the lumbar spine was determined by dual-energy X-ray absorptiometry (DEXA). RESULTS: Compared with age- and sex-matched control subjects, OP patients showed a 73% decrease in free IGF-I, a 29% decrease in total IGF-I, a 10% decrease in IGFBP-3, and a 52% decrease in IGFBP-5 levels; they had higher levels of IGFBP-1 (4.1-fold), IGFBP-2 (1.8-fold), IGFBP-4 (1.3-fold), and IGFBP-6 (2.1-fold). Alterations in IGF system components were most evident in 13 OP patients with vertebral fractures in the past 4 years compared to patients without fractures. In OP patients with fractures, the ratio between IGFBP-4 and IGFBP-5 was increased whereas levels of OSC were decreased. CONCLUSIONS: Our data provide strong indirect evidence for a functional connection between circulating IGF system components and bone metabolism and the susceptibility to fractures in OP patients. PMID- 12554010 TI - Halitosis in patients with Helicobacter pylori-positive non-ulcer dyspepsia: an indication for eradication therapy? AB - BACKGROUND: The aims of this study were to investigate the frequency of halitosis before and after eradication therapy and to determine whether halitosis is a valid indication for eradication therapy in patients with Helicobacter pylori (H. pylori)-positive non-ulcer dyspepsia. METHODS: Dyspepsia, related symptoms, and halitosis were investigated by way of a questionnaire. Only H. pylori-positive patients who showed no organic lesions on endoscopic examination and no atrophy histopathologically were included. A total of 148 patients fulfilled the above criteria and completed the study. Four weeks after the end of eradication treatment, the symptoms were re-evaluated and repeat endoscopy was done to check for H. pylori in the gastric mucosa. Results: H. pylori eradication was successful in 109 patients (73.6%). Prior to treatment, bloating was the most frequent symptom (74.3%), followed by diurnal pain (62.2%) and halitosis (61.5%). The most successfully resolved symptoms in the group as a whole, regardless of eradication status, were halitosis, diurnal pain, and hunger-like pain, respectively. In the patients with confirmed H. pylori eradication, the most successfully resolved symptoms were halitosis and hunger-like pain, respectively. CONCLUSION: Halitosis is a frequent, but treatable, symptom of H. pylori-positive non-ulcer dyspepsia and may be a valid indication for eradication therapy. PMID- 12554011 TI - B cell chronic lymphocytic leukemia with meningeal infiltration by T lymphocytes. AB - Chronic lymphocytic leukemia (CLL) is a generalized malignancy of the lymphoid tissue characterized by an accumulation of monoclonal lymphocytes, usually of the B cell type. Involvement of the central nervous system is a rare complication, usually seen in T cell leukemias. We report a case of a 78-year-old woman with B cell CLL and meningeal infiltration by both B and T lymphocytes, although predominantly T lymphocytes. Neurological symptoms were the first manifestation of this disease. Computed tomography of the brain was negative. The diagnosis of leukemic meningitis was made on the basis of the examination of the cerebrospinal fluid, and which included cytological and flow cytometry analysis. The patient was given systemic chemotherapy in the form of chlorambucil and intrathecal administration of methotrexate and dexamethasone. After recovery, she had regular follow-up. We assume that this rare case of CLL might have been biclonal, with both B and T cell types. PMID- 12554012 TI - Erdheim-Chester disease: a rare cause of paraplegia. AB - Erdheim-Chester disease (ECD) is a rare, non-Langerhans cell histiocytosis. It is characterized by osteosclerosis of the metaphyseal regions of long bones and several extraskeletal manifestations. Clinically, it ranges from an asymptomatic focal process to systemic disease with life-threatening visceral involvement. Until now, only two cases of Erdheim-Chester disease with paraparesis have been reported. Herein we report the first case of Erdheim-Chester disease with the clinical manifestation of paraplegia. Our patient also had diabetes insipidus, pleural and pericardial effusion, retro-orbital and cavernous sinus masses, fibrotic changes in the retroperitoneal, perirenal, and periaortic areas, and epidural space-occupying lesions. We want to emphasize that ECD may be a very rare cause of paraplegia. PMID- 12554013 TI - Severe thrombocytopenia secondary to cytomegalovirus infection in an immunocompetent adult. AB - A healthy 38-year-old man presented with a short history of a viral syndrome, accompanied by purpura and gingival bleeding. Examination of the blood showed a profound thrombocytopenia. Cytomegalovirus infection was diagnosed and a causal relationship between CMV infection and thrombocytopenia was assumed on clinical grounds. We successfully treated the patient with oral prednisone. In the discussion, we briefly mention the different hypothetical mechanisms leading to autoimmune thrombocytopenia. PMID- 12554014 TI - Recurrent pneumonia post-thymectomy as a manifestation of Good syndrome. AB - A 78-year-old woman presented with right lobar pneumonia. Six months before, the patient had undergone surgery due to a large mass that was found to be spindle cell thymoma. During the past 6 months, the patient had been admitted to the hospital several times with recurrent pneumonia in both lungs; this was treated successfully with antibiotics. Laboratory examinations revealed lymphopenia, eosinopenia, hypogammaglobulinemia, and a low count of CD4 T cells and an inverted CD4:CD8 T cell ratio, both on peripheral blood and bone marrow. History and laboratory findings were compatible with the diagnosis of the rare Good syndrome. This syndrome is discussed here. PMID- 12554015 TI - Hematogenous anaerobic vertebral osteomyelitis due to Bacteroides fragilis in a diabetic patient. AB - We report the case of a 70-year-old diabetic man with spontaneous vertebral osteomyelitis due to Bacteroides fragilis. Diagnosis was obtained on positive blood cultures. The port of entry remained unknown despite extensive investigation. A combination of metronidazole and clindamycin led to a clinical cure with no need for surgical debridement and no relapse after 9 months of follow-up. PMID- 12554016 TI - Hospitalists: the new model of inpatient medical care in the United States. PMID- 12554017 TI - Hospitalization for firearm-related injuries in the United States, 1997. AB - BACKGROUND: Firearm-related injuries are a serious public health problem in the United States. Despite the magnitude of this problem, prior national estimates of nonfatal, firearm-related morbidity have been limited to an emergency department based surveillance system. The objective of this study was to assess and report the information available on firearm-related injuries in an existing national database, derived from hospital discharge data. METHODS: Cross-sectional analysis of the 1997 Nationwide Inpatient Sample (NIS), a stratified probability sample of 1012 nonfederal community hospitals from 22 states. The database was queried using E codes to identify firearm-related injuries. The SUDAAN software program was used to convert raw counts into weighted counts that represent national estimates and 95% confidence intervals (CIs). RESULTS: An estimated 35,810 (95% CI, 32,615-38,947) cases nationwide were identified, of which 86% were male. Assault was the leading cause of firearm-related hospitalization, followed by unintentional injury. The mean length of stay (LOS) for patients with a firearm related assault was 6 days. Seven percent of all firearm cases died during the hospitalization. The total estimated hospital charges for firearm-related injuries in the United States in 1997 was over $802 million, and 29% of the patients admitted for this condition were uninsured. CONCLUSIONS: Firearm-related injuries rank highest among all conditions in the number of uninsured hospital stays, and the average LOS is much longer compared to other medical conditions. National estimates derived from the NIS are consistent with previous estimates, and NIS provides additional information not available from other data sources. PMID- 12554018 TI - Medical care utilization patterns in women with diagnosed domestic violence. AB - BACKGROUND: Information on women with domestic violence (DV) suggests increased healthcare utilization across all levels of care and many diagnoses. In the present ancillary study (1997-2002), derived from a large, group-randomized intervention trial we conducted in a staff-model health maintenance organization (HMO) (1995-1998), we examined total and incremental utilization rates, costs, and patterns for women aged >/=18 years with DV identified through the record reviews conducted for the trial. By the choice of comparison groups used, our present aim was to "bracket" any associated increase in utilization. METHODS: We compared visits and costs of medical-record confirmed cases of DV (n =62) to those for women without evidence of DV in the record (n =2287). These two groups were derived from women making visits for any one of four index reasons (injury, chronic pelvic pain, depression, or physical examination) associated with higher risk of DV or higher likelihood of its discussion. We constructed a second comparison group (n =6032) from the general population of enrolled women. We used the Chronic Disease Score to adjust for comorbidity. RESULTS: After adjusting for comorbidity, we found a 1.6-fold higher rate of all visits (95% confidence interval [CI]=1.4-1.9) and 1.6-fold higher estimated costs (95% CI=1.3-2.0) for abused women compared to non-DV women. The rates were 2.3-fold higher when compared to all enrolled women. CONCLUSIONS: Women with medical-record-documented DV demonstrate a pattern of increased utilization and costs across all levels of care and types of diagnoses. We conclude that being a DV case-patient is associated with between 1.6- and 2.3-fold increases in total utilization and costs. PMID- 12554019 TI - Do protection orders affect the likelihood of future partner violence and injury? AB - BACKGROUND: Approximately 20% of U.S. women who experience intimate partner violence (IPV) annually obtain a civil protection order (CPO). The effect of these orders on future abuse has been estimated in only a few studies, with mixed results. The objective of this study was to assess the effect of a CPO on the risk of future self-reported IPV and injury. METHODS: In this prospective cohort study of 448 adult female Seattle WA residents with IPV between October 1997 and December 1998, interviews were conducted at baseline, 5 months, and 9 months after the index incident. Odds ratios (ORs) estimated risks of contact; unwelcome calls or visits; threats; weapon threats; psychological, sexual, or physical abuse or injury; and abuse-related medical care among women who obtained a CPO after the index incident, compared with those who did not. RESULTS: Women who obtained a CPO following the index IPV incident had significantly decreased risk of contact by the abuser (OR=0.4); weapon threats (OR=0.03); injury (OR=0.3); and abuse-related medical care (OR=0.2) between the first and second follow-up interviews. Stronger decreases in risk were seen among women who had maintained the CPO throughout follow-up, which were significant for contact by the abuser (OR=0.2); weapon threats (OR=0.02); psychological abuse (OR=0.4); sexual abuse (OR=0.2); physical abuse (OR=0.3); injury (OR=0.1); and abuse-related medical care (OR=0.1) between first and second follow-up interviews. CONCLUSIONS: CPOs are associated with decreased likelihood of subsequent physical and nonphysical IPV. PMID- 12554020 TI - Adolescent participation in sports and adult physical activity. AB - BACKGROUND: Physical activity in adolescence has been reported to enhance physical activity in adulthood, but detailed information on the enhancing effect of different types of adolescent sports is lacking. We evaluated the association between participation in different types of adolescent sports and physical activity in adulthood. METHODS: The sample comprised 7794 males and females who responded to the mailed questions on physical activity status at age 14 years and at age 31 years in follow-up surveys of the Northern Finland 1966 birth cohort. The associations between adolescent participation in different sports and adult physical activity was examined by multinomial logistic regression. RESULTS: Frequent participation in sports after school hours in adolescence was associated with a high level of physical activity in adulthood. In males, adolescent participation in ball games, intensive endurance sports, track and field, and combat sports was associated with a high or very high level of adult activity. In females, the same applied to adolescent participation in running, orienteering, track and field, cycling, gymnastics, and riding. Adolescent participation in ball games increased participation in ball games in adulthood, especially in males, while participation in cross-country skiing, running, and orienteering provided the greatest stimulation to carry over of some endurance sport to adulthood. CONCLUSIONS: Participation in sports at least once a week among females and twice a week among males was associated with high level of physical activity in later life. Adolescent participation in the intensive endurance sports, and some sports that require and encourage diversified sports skills, appeared to be most beneficial with respect to the enhancement of adult physical activity. PMID- 12554021 TI - Trends in smoking before, during, and after pregnancy in ten states. AB - BACKGROUND: While trends in smoking prevalence during pregnancy are known, little is known about trends in quitting during pregnancy and resuming smoking after pregnancy. OBJECTIVES: This study examined the trends in and correlates of quitting during pregnancy and resuming smoking after pregnancy. METHODS: We used population-based random surveys of recent mothers in ten U.S. states (total of 115,000 women) conducted between 1993 and 1999. RESULTS: Although the prevalence of smoking 3 months before pregnancy was stable at around 26%, quitting during pregnancy rose from 37% to 46% between 1993 and 1999. Adjusted for maternal and state characteristics, the odds of quitting during pregnancy increased 51% between 1993 and 1999 (odds ratio [OR]=1.51; 95% confidence interval [CI]=1.08 2.12). Approximately half of the women who quit smoking during pregnancy resumed smoking within 6 months postpartum. Primiparous, privately insured, college educated women are more likely to quit and least likely to resume smoking after delivery, compared to multiparous, Medicaid-insured, and high school-educated women. Teenaged women are more likely to quit, but also more likely to resume smoking than older women. CONCLUSIONS: The increase in quit rates during pregnancy is encouraging, but the lack of any change in smoking before pregnancy or in postpartum relapse rates suggests that permanent changes in maternal smoking will require additional focus. PMID- 12554022 TI - Chlamydia trachomatis infection in asymptomatic men. AB - BACKGROUND: The epidemiology of Chlamydia trachomatis infection in men is not well defined, especially among those who are asymptomatic or show no signs of infection. Established C. trachomatis screening programs for women have demonstrated the benefit of routine screening in reducing prevalence over time, but the yield and benefit of screening asymptomatic men are unclear. METHODS: Cross-sectional study of C. trachomatis prevalence and associated risk factors among men tested at sexually transmitted disease (STD) clinics in Alaska, Idaho, Oregon, and Washington. We analyzed data from 43,094 men universally tested from 1997 to 1999 at 103 STD clinics, and assessed age-specific prevalence of infection, controlling for signs of infection (urethritis diagnosed by clinician) and report of sexual contact to a person with an STD (defined as "contact"). RESULTS: Overall prevalence of C. trachomatis was 10.3%. Age-specific prevalence was highest among men aged 18 to 19 years and lowest among those aged >29 years, regardless of signs of infection upon examination or contact to a person with an STD. If these factors and age <25 years had been used to direct C. trachomatis testing at STD clinics, 59% of men would have been tested and 91% of positives would have been detected. CONCLUSIONS: Using either the presence of clinical signs or report of a sex partner with an STD in combination with selective screening of all men aged <25 years detects the majority of infections and, in our population, would have considerably reduced the number of negative tests performed. PMID- 12554023 TI - Dietary supplement use and medical conditions: the VITAL study. AB - BACKGROUND: Over half of U.S. adults use vitamin or mineral supplements, and some are likely using supplements to treat chronic diseases or risk factors for disease. Information on the relationship between supplement use and medical conditions is useful to health professionals to understand the self-medication behavior of their patients, and important for researchers because medical conditions may be potential confounding factors in observational studies of supplement use and disease risk. METHODS: The cross-sectional data in this report are from 45,748 participants, aged 50 to 75 years, who completed a self administered, mailed questionnaire on current dietary supplement use (multivitamins plus 16 individual vitamins or minerals), medical history (cancer, cardiovascular-related diseases, and other self-reported medical conditions), and demographic characteristics. RESULTS: Supplement use (mean number used at least once a week) was higher among respondents who were older, female, highly educated, Caucasian, and of normal body mass index (all p<0.001). After controlling for these covariates, supplement use was higher among those with the condition for 13 of the 21 conditions examined (p<0.01); only having diabetes or high stress was associated with using fewer supplements. For specific supplements, the strongest associations were for cardiovascular disease and its risk factors with vitamin E, niacin, and folate, and for calcium with indigestion and acid reflux disease. For several conditions, the relative odds of using specific supplements were consistently higher for men than for women. CONCLUSIONS: Supplement use was associated with many medical conditions in this cohort. However, these cross-sectional data do not permit inferences about the temporal sequence. Some associations appeared to be based on evidence for efficacy (e.g., folate with coronary artery disease), and others could be based on misinformation (e.g., selenium with benign prostatic hyperplasia). PMID- 12554024 TI - Squire's Quest! Dietary outcome evaluation of a multimedia game. AB - BACKGROUND: Fruit, juice, and vegetable (FJV) consumption among children is low. Innovative programs are needed to enable children to increase FJV intake. Psychoeducational multimedia permits the delivery of interventions as designed and capitalizes on known behavior change principles. DESIGN: Elementary school was the unit of recruitment, assignment, and analysis. Twenty-six elementary schools were pair matched on size and percentage of free or reduced-price lunch, and randomly assigned to treatment or control groups. Data were collected just before and just after the program. SETTING/PARTICIPANTS: All fourth-grade students in participating elementary schools were invited to participate. Data were collected on 1578 students. MAIN OUTCOME Servings of fruit, 100% juice, and vegetables consumed. INTERVENTION: Squire's Quest! is a ten-session, psychoeducational, multimedia game delivered over 5 weeks, with each session lasting about 25 minutes. Based on social cognitive theory, educational activities attempted to increase preferences for FJV through multiple exposures and associating fun with their consumption, increase asking behaviors for FJV at home and while eating out, and increase skills in FJV preparation through making virtual recipes. MEASURES: Four days of dietary intake were assessed before and after the intervention. Assessment was made by the Food Intake Recording Software System (FIRSSt), which conducts a multiple pass, 24-hour dietary intake interview directly with the children. RESULTS: Children participating in Squire's Quest! increased their FJV consumption by 1.0 servings more than the children not receiving the program. CONCLUSIONS: Psychoeducational multimedia games have the potential to substantially change dietary behavior. More research is warranted. PMID- 12554025 TI - Health plan quality-of-care information is undermined by voluntary reporting. AB - BACKGROUND: Most health services in the United States are delivered by managed care organizations (MCOs). Publicly available, plan-specific performance information is required to adequately assess healthcare quality provided. Using women's health indicators, we compared performance results for MCOs and evaluated whether those MCOs that publicly report quality-of-care (QOC) results demonstrate better QOC than those plans that restrict public access to data. METHODS: Data from the Health Plan Employer Data and Information Set (HEDIS) for commercial MCOs in 1998 were analyzed for women's QOC indicators. Plan-specific, regional, and national performances were analyzed and results compared to established benchmarks. Public-reporting plans were compared to plans that restrict access to QOC information. Linear regression was used to identify determinants of health plan performance including public release of information. RESULTS: Commercial MCOs had wide variations in QOC indicators and, on average, failed to attain national health goals for most women's health indicators analyzed. Plans that restricted public access to QOC information had poorer performance than those that did not (p<0.05). Results suggest that whether a plan publicly releases its performance information is highly associated with health plan performance even after taking into account other factors. CONCLUSIONS: The voluntary aspect of reporting and the ability of health plans to restrict public access is allowing poorer performing health plans to escape public scrutiny. Variations in QOC have clinical significance and, if publicly available, would enable individuals to select high-quality healthcare products. The ability of health plans to restrict public information is not consistent with the 1973 Health Maintenance Organization Act requiring public information on health plan quality. A national strategy to ensure that QOC information is available on all healthcare systems is past due. PMID- 12554026 TI - Physician advice about smoking and drinking: are U.S. adults being informed? AB - BACKGROUND: Population-based estimates for the prevalence of smokers receiving advice from a health professional to quit smoking and the prevalence of binge drinkers being talked to about alcohol use are lacking for U.S. adults. This information is useful for clinicians and public health professionals. METHODS: Data are from the Behavioral Risk Factor Surveillance System, a continuous random digit-dial telephone survey of U.S. adults. In 1997, 10 states collected data on these health interventions for tobacco and alcohol use. The prevalence of professional advice to quit smoking and about alcohol use was calculated and examined by demographic characteristics. The number of at-risk adults who had a routine checkup in the last year and had not received these interventions was also estimated. RESULTS: By self-report, 70% of smokers were advised to quit, and 23% of binge drinkers were talked to about their alcohol use. Using multivariate logistic regression analyses, we found among smokers that women and older persons were more likely to receive advice; among binge drinkers, health intervention was more likely to occur for men and non-Hispanic blacks. Across the 10 states, approximately 2 million smokers and 2 million binge drinkers with a routine checkup in the past 12 months were not advised to quit smoking or talked to about their alcohol use. CONCLUSIONS: Many opportunities to intervene with smokers and binge drinkers are lost. Efforts to increase physician education and to identify and reduce other barriers may help. PMID- 12554027 TI - Counseling to promote a healthy diet in adults: a summary of the evidence for the U.S. Preventive Services Task Force. AB - OBJECTIVE: The purpose of this study was to examine the effectiveness of counseling to promote a healthy diet among patients in primary care settings. DESIGN AND DATA SOURCES: We conducted a MEDLINE search from 1966 to December 2001. STUDY SELECTION: We included randomized controlled trials of at least 3 months' duration with measures of dietary behavior that were conducted in patient populations similar to those found in primary care practices. We excluded studies that reported only biochemical or anthropomorphic endpoints, had dropout rates greater than 50%, or enrolled patients based on the presence of a chronic disease. DATA EXTRACTION: One author extracted relevant data from each included article into evidence tables. Using definitions developed by the research team, two authors independently rated each study in terms of its effect size, the intensity of its intervention, the patient risk level, and the use of well-proven counseling techniques. DATA SYNTHESIS: We identified 21 trials for use in this review. Dietary counseling produces modest changes in self-reported consumption of saturated fat, fruits and vegetables, and possibly dietary fiber. More intensive interventions were more likely to produce important changes than brief interventions, but they may be more difficult to apply to typical primary care patients. Interventions using interactive health communications, including computer-generated telephone or mail messages, can also produce moderate dietary changes. CONCLUSIONS: Moderate- or high-intensity counseling interventions, including use of interactive health communication tools, can reduce consumption of saturated fat and increase intake of fruit and vegetable. Brief counseling of unselected patients by primary care providers appears to produce small changes in dietary behavior, but its effect on health outcomes is unclear. PMID- 12554028 TI - Behavioral counseling in primary care to promote a healthy diet: recommendations and rationale. PMID- 12554029 TI - Education of preventive medicine residents: alcohol, tobacco, and other drug abuse. AB - BACKGROUND: Generalist physicians' addiction training is inadequate, but general preventive medicine residency (PMR) programs have not been studied. We determined PMR programs' alcohol, tobacco, and other drug abuse (ATOD) training from 1995 to 2000 and identified barriers to this education. METHODS: Interviewer-administered telephone survey of program directors (PDs) of accredited PMR programs in the United States. RESULTS: We interviewed all 41 PMR PDs. While 78% of PMR PDs reported interest in increasing ATOD education, for 68% it was not a high educational priority. Tobacco ranked in the top third of preventive medicine topics by 58%, while alcohol and other drugs ranked in the bottom third by 48% and 52%, respectively. Twenty-two percent of programs required a clinical ATOD rotation, most commonly smoking-cessation clinics. Only 29% of PMR PDs felt that residents were well prepared in clinical aspects of ATOD, while 60% felt that residents were prepared in ATOD research and public health issues. The most commonly reported barriers to ATOD training were lack of resident interest and defined competencies (64% each); limited faculty time (59%); limited teaching time (54%); lack of available teaching materials (53%); and lack of faculty expertise (51%). CONCLUSIONS: While the majority of PMR PDs recognize the importance of incorporating teaching about addictions into training, much of the ATOD education in PMRs focuses on tobacco alone. Setting educational standards, defining competencies, investing in faculty development, and creating ATOD curricular modules are important next steps toward preparing preventive medicine physicians to effectively reduce the public health toll of addictions. PMID- 12554031 TI - Science and the advancement of eHealth: a call to action. PMID- 12554032 TI - Prospective evaluation of criteria for obtaining thoracolumbar radiographs in trauma patients. AB - This study examined if use of clinical screening criteria for selective radiography of blunt trauma patients can identify all patients with thoracolumbar (TL) spine injuries. The study was a prospective cohort of patients undergoing TL spine radiographs following blunt trauma. Patients were considered at risk for TL spine injury if they had any of the following clinical criteria: 1) complaints of TL spine pain, 2) TL spine tenderness, 3) a decreased level of consciousness, 4) intoxication with ethanol or drugs, 5) a neurologic deficit, or 6) a painful distracting injury. Patients without any of these findings were considered at low risk for TL spine injury. Severity of mechanism of injury was also recorded. Data sheets were completed prior to TL radiography. Injury status was determined by the final faculty radiologist interpretation of all radiographic studies. A total of 2404 patients were enrolled. TL spine injuries were identified in 152 patients. Of these 152 patients with spine injuries, all 152 (100%, 95% confidence interval 98-100%) were considered high risk by having at least one of the high-risk criteria. These criteria have a specificity of 3.9%, a positive predictive value of 6.6%, and a negative predictive value of 100%. All of the high-risk criteria but intoxication with ethanol or drugs were important as sole predictors of TL spine injury. The use of high-risk clinical screening criteria identified virtually all blunt trauma patients with acute TL spine injuries. These criteria, however, have poor specificity and positive predictive value. PMID- 12554033 TI - Coronary angiographic findings in patients with cocaine-associated chest pain. AB - Patients who present to the Emergency Department (ED) with chest pain associated with cocaine use are a common problem. The incidence and predictors of underlying significant coronary disease in patients with and without myocardial infarction (MI) has not been well described. Patients who underwent coronary angiography within 5 weeks of an ED evaluation for cocaine-associated chest pain were studied. Significant disease was defined as > or = 50% stenosis of a coronary artery or major branches or bypass graft. A total of 90 patients underwent coronary angiography. Significant disease was present in 45 (50%), with 1-vessel disease in 32%, 2-vessel disease in 10%, 3-vessel disease in 6%, with significant graft stenosis in 3%. Significant disease was present in 77% of patients with MI or troponin I elevations, compared to only 35% of patients without myonecrosis. Predictors of significant coronary disease included MI or troponin I elevations, prior MI, known coronary disease (prior MI or revascularization), and elevated cholesterol. Only 7 of the 39 patients (18%) without myonecrosis or a history of coronary disease had significant disease on angiography. In conclusion, significant disease is found in the majority of patients with cocaine-associated MI or troponin elevations. In contrast, only a minority of those without myonecrosis have significant coronary disease. PMID- 12554034 TI - Odontoid hypoplasia presenting as torticollis: a discussion of its significance. AB - Odontoid dysplasias are considered rare but are becoming increasingly recognized. Patients may have no symptoms, localized neck pain, or neurologic symptoms. Because patients with odontoid anomalies have the potential for craniovertebral instability, recognition of the entity is essential. A discussion of the axis, its development and anomalies follows. PMID- 12554035 TI - Bilateral internuclear ophthalmoplegia following minor head trauma. AB - Internuclear ophthalmoplegia (INO) is characterized by pathognomonic findings on neurological examination. It results from a lesion in the medial longitudinal fasciculus (MLF) and is rarely caused by head trauma. The neuroanatomy of INO is complex and the mechanism by which trauma causes this syndrome is controversial. In the context of trauma, INO occurs frequently in association with other neurological findings and should prompt a thorough investigation and ICU admission. A case of an individual with acute post-traumatic INO is reported and discussed. PMID- 12554036 TI - Reexpansion pulmonary edema: a case report and review of the current literature. AB - This case report illustrates the presentation and course of reexpansion pulmonary edema (REPE) in a young man with spontaneous pneumothorax. REPE is considered relatively uncommon by most accounts, but in certain clinical circumstances the incidence is much higher. Although supportive therapy is the rule, the condition is far from benign and mortality estimates are as high as 20%. Risk factors, including young age, a large pneumothorax and longer duration of collapse, may help predict which patient will encounter this complication. In patients with these risk factors, the thoracostomy tube should be initially left off suction in an effort to prevent REPE primarily. When REPE is encountered, therapy is supportive. PMID- 12554037 TI - Ventricular tachycardia secondary to Port-a-Cath fracture and embolization. AB - Distal embolization of a fractured indwelling venous access catheter is a known, but rare, complication of these common devices. We report a case of possibly life threatening tachydysrhythmia caused by an embolized portion of catheter from a totally implanted venous access catheter. Our discussion focuses on review of the literature on the typical presentation and management of this entity, as well as a brief discussion on prevention and risk factors for developing this complication. PMID- 12554038 TI - Electrocardiographic manifestations: narrow QRS complex tachycardias. AB - Narrow QRS complex tachycardia is a common dysrhythmia in Emergency Medicine practice. Diagnosis and mechanism often can be made by 12-lead electrocardiographic (EKG) analysis but may subsequently require electrophysiologic testing. The clinical manifestations are varied and dependent upon heart rate, prior cardiac disease, and general physiologic status. Patient management is directed towards the etiology and mechanism of the dysrhythmia and includes vagal maneuvers, pharmacologic therapy, and cardioversion. Hemodynamically compromised patients must be promptly treated. Patients are often admitted to the hospital but selected patients can be safely discharged from the Emergency Department for outpatient evaluation and management. Pediatric and pregnant patients are, in general, treated the same as adults. Several case examples and EKGs are presented. PMID- 12554039 TI - Utility of platelet adp receptor antagonism in the emergency department: a review. AB - Emergency physicians are often the first clinicians to evaluate patients with acute atherothrombotic events. Platelet adenosine diphosphate (ADP) receptor antagonists, by specifically and irreversibly blocking ADP-induced platelet activation and aggregation, may reduce the injury associated with this process and can prevent recurrent ischemic events. Their role in the prevention of recurrent vascular events has been well documented. Recently, the CURE Trial showed that the combination of aspirin and clopidogrel improved outcomes in patients with non-ST-segment-elevation acute coronary syndrome (ACS). Familiarity with ADP receptor antagonists and knowledge about their appropriate use is important to the emergency physician in the management of ACS and potentially in that of transient ischemic attacks (TIAs), ischemic strokes, and acute peripheral arterial obstruction. This review addresses the pathophysiology of atherothrombosis and evaluates the potential use of ADP receptor antagonists in the Emergency Department setting. PMID- 12554040 TI - Abdominal pain in first trimester pregnancy. PMID- 12554041 TI - Great toe pain. PMID- 12554042 TI - Bilateral luxatio erecta. PMID- 12554043 TI - Iliac artery aneurysm presenting with lower extremity deep vein thrombosis. PMID- 12554044 TI - Original descriptions of the classic signs of aortic valve insufficiency. AB - The valves of the heart and their diseases have generated a great deal of interest since ancient times. The early observations regarding valvular deformities were confined to the morbid changes with no reference to the hemodynamic significance. The classic clinical signs of aortic valve insufficiency indicating hemodynamic disturbances were described later. The historical background and original descriptions of these classic signs of aortic valve insufficiency are presented. PMID- 12554045 TI - Personal and literary experiences in the development of an emergency physician. AB - The development of Emergency Medicine in the United States has evolved in diverse manners, depending particularly on the societal needs and academic support available in each city and institution. Significant personal and literary experiences are described with regard to the author's role in the development of Emergency Medicine at Bellevue Hospital Center and the New York City Poison Center in New York City. Critical quotations of literary and philosophic importance for the author are utilized to demonstrate the importance of literature in the development of a philosophy of life, a residency program in Emergency Medicine, a residency program in Medical Toxicology and an academic Department of Emergency Medicine. Critical societal events during the last quarter of the 20(th) century are related to this personal formative literature and the development of Emergency Medicine. PMID- 12554046 TI - A better imaging alternative. PMID- 12554047 TI - Futility in resuscitation from cardiac arrest: role of out-of-hospital healthcare professionals. PMID- 12554048 TI - A fatal case of toxic epidermal necrolysis associated with liver cirrhosis. PMID- 12554049 TI - The National Emergency Medical Extranet project. AB - The National Emergency Medical Extranet (NEME) project was a collaborative multi center effort to create a plan for a networked system to improve emergency clinical care through real-time information support, and simultaneously provide benefit through information support for public health initiatives. This article presents a review of the NEME project and its recommendations, which are particularly relevant given the desire for improved communication and surveillance systems in today's healthcare and public health environments. Participants in the NEME project performed an environmental assessment and a proposed conceptual architecture for NEME. A consensus conference was held to review the NEME concept to obtain feedback and delineate priorities for future development and testing. The NEME consensus conference used a modified version of the nominal group method. Recommendations for the following areas were established: Business/Organizational Issues [1) create a compelling provider driven NEME model, 2) provide a comprehensive policy framework, 3) address economics]; Clinical/Caregiver Issues [1) develop a NEME system that is integrated with Emergency Medicine workflow, 2) provide incentives to caregivers, 3) generate a critical mass of participation for maximum benefit]; Technical Issues [1) incorporate a robust security and confidentiality architecture, 2) utilize a master person index, where appropriate, 3) evaluate or adopt existing data standards]; Heart Attack Alert Functional Priorities for NEME [1) continuous quality improvement and research, 2) regional electrocardiogram server, 3) past medical history and medication server]; Next Generation Internet Functional Priorities for NEME [1) real time epidemiology/surveillance, 2) patient education, 3) real time clinical alerting]. In conclusion, issues and consensus recommendations in the planning of a NEME are documented. These recommendations should be considered in future efforts to design, develop and implement wide area information networks to support Emergency Medicine. A review of current activities evolving from NEME is presented, and further research and development is encouraged to create and implement NEME systems. PMID- 12554050 TI - Prehospital resuscitation practices: a survey of prehospital providers. AB - Numerous factors affect decision making in the prehospital provision of resuscitative care. This study was undertaken to determine current practices involved in the initiation, continuation and termination of resuscitative efforts, and the impact of advance directives, in the prehospital resuscitation setting. This cross-sectional mailed questionnaire surveyed 3807 members of the National Association of Emergency Medical Technicians. The study instrument included questions regarding the termination and withholding of resuscitative efforts in the prehospital setting, as well as survival rates, local protocols and compliance with advance directives. Of 1546 respondents (41% response rate), with a mean 9.0 years of experience, most (89%) indicated that they would withhold resuscitative efforts in the presence of an official state-approved advance directive. However, very few providers would withhold resuscitative efforts if only an unofficial document (4%) or verbal report of an advance directive (10%) were available. Providers with more than 10 years experience were more likely to withhold resuscitation attempts in the presence of only a verbal report of an advance directive (p = 0.02, Chi-square), and were more likely to withhold resuscitation attempts in situations they considered futile (p = 0.001, Chi-square). Most (77%) respondents have local EMS guidelines for termination of resuscitation in the prehospital setting, but 23% of those consider existing guidelines to be inadequate. The majority of prehospital providers stated that they honor official state-approved advance directives, but do not follow directives from unofficial documents or verbal reports of advance directives. More experienced providers stated that they withhold resuscitative efforts more often in futile situations, or in the presence of unofficial advance directives. Advance directives should be utilized more uniformly among patients who wish to forgo resuscitative efforts in the event of cardiac arrest. Because many local protocols are judged to be inadequate, we support the institution of improved clinical guidelines regarding the prehospital termination of resuscitative efforts. PMID- 12554051 TI - Late rebound digoxin toxicity after digoxin-specific antibody Fab fragments therapy in anuric patient. PMID- 12554053 TI - Towards complete and accurate reporting of studies of diagnostic accuracy: the STARD initiative. AB - OBJECTIVE: To improve the accuracy and completeness of reporting of studies of diagnostic accuracy in order to allow readers to assess the potential for bias in the study and to evaluate its generalisability. METHODS: The Standards for Reporting of Diagnostic Accuracy (STARD) steering committee searched the literature to identify publications on the appropriate conduct and reporting of diagnostic studies and extracted potential items into an extensive list. Researchers, editors, and members of professional organizations shortened this list during a two-day consensus meeting with the goal of developing a checklist and a generic flow diagram for studies of diagnostic accuracy. RESULTS: The search for published guidelines regarding diagnostic research yielded 33 previously published checklists, from which we extracted a list of 75 potential items. At the consensus meeting, participants shortened the list to 25 items, using evidence on bias whenever available. A prototypical flow diagram provides information about the method of patient recruitment, the order of test execution and the numbers of patients undergoing the test under evaluation, the reference standard or both. CONCLUSIONS: Evaluation of research depends on complete and accurate reporting. If medical journals adopt the checklist and the flow diagram, the quality of reporting of studies of diagnostic accuracy should improve to the advantage of the clinicians, researchers, reviewers, journals, and the public. PMID- 12554054 TI - Biochemical markers of alcohol use in pregnant women. AB - OBJECTIVES: To describe the serious health consequences of alcohol (ethanol) use, especially as they relate to pregnancy and the development of fetal alcohol syndrome (FAS) and fetal alcohol effects (FAE). The classic markers of alcohol exposure, including blood/breath alcohol, gamma-glutamyl transferase (gammaGT), mean corpuscular volume (MCV), hemoglobin-associated acetaldehyde (HAA) and carbohydrate deficient transferrin (CDT), are valuable and their methods of analysis are reviewed. CONCLUSIONS: Since both FAS and FAE represent two of the leading preventable causes of mental retardation and birth defects, identification of alcohol use early in pregnancy is important to avoid adverse fetal outcomes. Unfortunately, the diagnosis of FAS and FAE is usually made after birth, when alcohol damage has become irreversible and permanent. The clinical laboratory can help prevent this damage and make a valuable contribution in assessing prenatal alcohol use. The clinical utility of blood/breath alcohol, gammaGT, MCV, HAA and CDT in alcohol use identification, especially in pregnancy, is substantial. Although none of the markers singularly has adequate sensitivity and specificity for screening, their diagnostic utility increases when measured as a panel. This is especially true in detecting alcohol use in pregnancy where the presence of several positive markers was correlated with the presence of alcohol-related fetal effects. PMID- 12554056 TI - Evaluation of point-of-care test systems using the new definition of myocardial infarction. AB - OBJECTIVES: To examine the proportion of patients admitted to CCU because of chest pain with a negative quantitative troponin t-test (Cardiac reader; Roche), who despite the negative test, would fulfil the new myocardial infarction criteria. A second aim was to evaluate the clinical utility of troponin I. DESIGN AND METHODS: Troponin T (Cardiac reader; Roche) was measured 12 h after the last chest pain episode. If Troponin T was negative the subjects were included in the study. All included subjects were also examined with troponin I (Stratus CS; Dade Behring) and troponin T at the central laboratory (Elecsys 2010) at the same time. 187 patients were included. CKMB was also measured 6 h after the last chest pain (Stratus CS; Dade Behring). RESULTS: Fifteen patients (8.0%) fulfilled the criteria of myocardial infarction, despite a negative Troponin T (Cardiac reader; Roche). CKMB measures did not add useful diagnostic information. The sensitivity and specificity were 100% and 95.3%, respectively for troponin I, when 0.2 microg/L was used as cut off level for myocardial infarction. CONCLUSION: In this low risk group, eight percent of the patients with a negative Troponin T (Cardiac reader; Roche) fulfilled the new criteria of myocardial infarction. Troponin I (Stratus CS; Dade Behring) appeared to be a reliable method in this group. PMID- 12554055 TI - Pediatric reference ranges for zinc protoporphyrin. AB - OBJECTIVE: To determine pediatric reference ranges for zinc protoporphyrin (ZPP). DESIGN AND METHODS: The study was conducted in a large pediatric hospital on patient blood specimens (n = 6,478) (0-17 y) accrued from January 2001 to June 2002. The data was analyzed employing the Hoffman approach, which was computer adapted. RESULTS AND CONCLUSIONS: The 2.5(th) and 97.5(th) percentiles for children age 0 to 12 months were 9 to 40 microg/dL (16.6-73.6 micromol/mol heme) for female subjects and 8.5 to 34.5 microg/dL (15.6-63.5 micromol/mol heme) for males. The 97.5(th) percentiles decreased for the 13 to 24 months age group for females (32 microg/dL) (58.9 micromol/mol heme). There was a significant decrease in the 97.5(th) percentile for zinc protoporphyrin (ZPP) concentrations for the 5 to 9 yr age group, the 97.5(th) percentile being 30 microg/dL (55.2 micromol/mol heme) in both genders, which increased to 33.5 microg/dL (61.6 micromol/mol heme) in the 10 to 17 yr female age group but not for the males (31.5 microg/dL) (58.0 micromol/mol heme). The highest medians were 25.5 microg/dL (46.9 micromol/mol heme) for females, and 21.5 microg/dL (39.6 micromol/mol heme) for males in the 0 to 12 months age group. PMID- 12554057 TI - Lipase and pancreatic amylase versus total amylase as biomarkers of pancreatitis: an analytical investigation. AB - OBJECTIVE: To evaluate the biomarkers of pancreatitis Colorimetric Lipase, Total Amylase and Pancreatic Amylase (immunoinhibition) assays on the Roche COBAS INTEGRA 700. RESULTS: Pancreatic and Total Amylase assays and Colorimetric Lipase showed excellent imprecision of 1.6 to 2.3% and linearity (slope = 0.94-0.99, y intercepts-1 to +3 U/L, r = 0.999) over the range of 17 to 900, 35 to 880, and 21 to 150 U/L, respectively. There was an excellent correlation between Pancreatic and Total Amylase: Pancreatic Amylase = 0.99 (+/- 0.02) x Total Amylase-36(+/- 8) (n = 106, r = 0.97, p < 1 x 10(-5), y intercept p < 1 x 10(-5)). Colorimetric Lipase showed some correlation to Total and Pancreatic Amylase results: Colorimetric Lipase = 1.54 (+/- 0.16) x Total Amylase-81(+/- 37) (n = 100, r = 0.70, p < 1 x 10(-6), y intercept p = 0.03), and Colorimetric Lipase = 1.78 (+/- 0.15) x Pancreatic Amylase-50(+/- 29) (n = 99, r = 0.78, p < 1 x 10(-6), y intercept p = 0.09). CONCLUSION: We recommend running the more specific Pancreatic Amylase as biomarker of pancreatitis on the Roche COBAS INTEGRA. PMID- 12554058 TI - Microfluidic chip-based method for genotyping microsatellites, VNTRs and insertion/deletion polymorphisms. AB - We have developed a method to genotype variable number of tandem repeats (VNTRs) and insertion/deletion polymorphisms using an integrated microfluidic chip-based system. We used this method to analyze a) a highly polymorphic pentanucleotide repeat (CCTTT)(n) locus within the 5'-putative promoter region of the human inducible nitric oxide synthase gene (iNOS5) which is associated with diabetic complications and infectious diseases; b) a bi-allelic 27 bp VNTR region within intron 4 of endothelial nitric oxide gene (eNOS27) which is associated with hypertension in type 2 diabetes patients with coronary heart disease and excess risk of advanced diabetic nephropathy in type 1 diabetes patients and c) an insertion/deletion polymorphism within the gene encoding angiotensin-converting enzyme (ACE/ID) which is associated with cardiovascular pathology and nitric oxide activity, and is in strong linkage disequilibrium with functional variants. Following amplifications, samples were mixed with gel-dye and markers and loaded into commercially available microfluidic chips designed for DNA sizing applications. In the study (N = 230), 95 (41%) of the DNA samples were homozygous and 135 (59%) were heterozygous for the iNOS5 repeats. For eNOS27, 173 (75%) of the genotyped DNA samples were homozygous for the larger 4b allele and the remaining 57 samples (25%) were heterozygous (4b/4a). No DNA samples were homozygous for the shorter 4a allele with four 27 bp repeats. In case of ACE/ID, 47 (20%) of the DNA samples were homozygous for the insertion, 65 (28%) were homozygous for the deletion and the remaining 118 (51%) were heterozygous. The results obtained were verified by analyzing random amplicons using bi-directional sequencing and GeneScan 3.0 analyses with 100% concordance being observed. Using the microfluidic chip-based method, separation and DNA sizing and genotyping are rapidly accomplished. The DNA fragments are resolved clearly and the system allows quantitation. Finally, the microfluidic chip-based method may be used for both large- and small-scale genotyping studies. PMID- 12554059 TI - Antigenicity of a recombinant NS3 protein representative of ATPase/helicase domain from hepatitis C virus. AB - It has been shown that the Hepatitis C virus nonstructural NS3 protein possesses at least two enzymatic domains: a serine-protease domain and an adenosine triphosphatase (ATPase)/helicase domain. In this report, a truncated fragment of NS3 (26 kDa), representing main epitopes from the (ATPase)/helicase domain, has been expressed in Escherichia coli. The recombinant protein was purified by Ion Metal Affinity Chromatography (IMAC) with more than 90% purity. The recognition of B-cell linear epitopes in the NS3 protein was evaluated by immunoblot. The recombinant NS3 protein was reduced and carboxymethylated, and the recognition of either conformational and/or linear B-cell determinants was evaluated by ELISA. The inclusion of the recombinant NS3 protein in a third-generation diagnostic system UltraMicroELISA (UMELISA) allowed an increase in the sensitivity, due to the detection of a new variety of false-negative sera in blood donor test samples. PMID- 12554060 TI - Relation of aging with oxidative protein damage parameters in the rat skeletal muscle. AB - OBJECTIVES: An increase in oxidative stress may contribute to the development of oxidative protein damage in the aging rat skeletal muscle. Our aim was to reveal protein carbonyl (PCO), advanced oxidation protein products (AOPP), a novel marker of oxidative stress, and protein thiol (P-SH) levels as markers of protein oxidation, as well as lipid hydroperoxide (LHP) levels as a marker of lipid peroxidation, and relation of nitrotyrosine (NT) levels with these markers in skeletal muscle tissue of young, adult, and old male Wistar rats. DESIGN AND METHODS: In the present study, we investigated the relation between aging and oxidative protein damage parameters such as PCO, NT, AOPP, and P-SH, as well as oxidative stress parameters such as total thiol, nonprotein thiol, and LHP in the skeletal muscle tissue of young, adult, and old Wistar rats. RESULTS: PCO and NT levels of old rats were significantly increased compared with those of young and adult rats. Skeletal muscle AOPP levels were significantly increased in old rats compared with those of adult rats. P-SH levels were significantly decreased in old rats compared with those of young rats. CONCLUSIONS: The finding that the increase in PCO levels of young vs. old group was more significant than that of adult vs. old group may suggest that PCO formation is an early specific marker of aging process in skeletal muscle. In addition, increased levels of nitrotyrosine in the skeletal muscle of the old rat group may be a novel specific marker of oxidative protein damage in the aging muscle. The absence of correlation between oxidative protein damage markers mentioned above and LHP levels may indicate that protein oxidation and lipid peroxidation in the aging rat skeletal tissue are two distinct mechanisms. PMID- 12554061 TI - Peripheral-type benzodiazepine receptors in human mononuclear cells of patients affected by osteoarthritis, rheumatoid arthritis or psoriasic arthritis. AB - OBJECTIVES: The objective of this study was to evaluate the kinetic parameters at equilibrium of peripheral benzodiazepine receptors (PBR) in human mononuclear cells from patients affected by osteoarthritis (OA), rheumatoid arthritis (RA) and psoriasic arthritis (PA). DESIGN AND METHODS: Mononuclear cells were obtained from 10 patients with OA, 10 patients with RA and 10 patients with PA. Evaluation of kinetic parameters of PBR was performed using [(3)H]PK 11195, a specific radioligand for this receptor, and compared with 10 healthy controls. RESULTS: The results show a statistically significant decrease (37.5%, as an absolute percentage) in the maximal number of binding sites (B(max)) of patients with OA, compared with healthy controls; however, the values of the dissociation constant (K(d)) at equilibrium do not show any statistically significant variations. CONCLUSIONS: These data further confirm the presence of peripheral biochemical alterations in OA. As peripheral benzodiazepine receptors appear to be involved in the immune function, and in the protection of hematopoietic cells against oxygen radical damage, the observed decrease in B(max) might be related to cellular protection. PMID- 12554062 TI - Biochemical changes in tumor tissues of oral cancer patients. AB - OBJECTIVES: Our aim was to assess the oxidative stress in tumor tissues of oral cancer patients by measuring the levels of lipids, thiobarbituric acid reactive substances (TBARS) and antioxidants. DESIGN AND METHODS: This study has been conducted on oral tumor tissues of 48 adult male oral cancer patients with various clinical stages (stage II to stage IV; 16 of each) and normal tissues of an equal number of age and sex matched disease free healthy subjects. The levels of lipids, lipid peroxidation and antioxidants in buccal mucosa of healthy subjects and oral cancer patients were assayed using colorimetric methods, except for vitamin E (fluorometriy). RESULTS: Total cholesterol and esterified cholesterol were markedly increased whereas phospholipids and free fatty acids were significantly decreased in tumor tissues as compared to normal tissues. The decrease in TBARS levels and functional compromise of antioxidant defense mechanisms were observed in the tumor tissues as compared to normal subjects. CONCLUSIONS: The low availability of peroxidizable substrates and the enhanced antioxidant capacity in tumor tissues make them less susceptible to oxidative stress, conferring a selective growth advantage on tumor cells. PMID- 12554063 TI - The effects of anesthetic techniques on acute phase response at delivery (anesthesia and acute phase response). AB - OBJECTIVES: To investigate the effects of anesthetic techniques and delivery types on stress response during uncomplicated delivery. DESIGN AND METHODS: Forty pregnant women at term were divided into four groups. Group-I, cesarean section with general anesthesia; Group-II, section with spinal anesthesia; Group-III, section with epidural anesthesia; and Group-IV, vaginal delivery with epidural analgesia. C-reactive protein and albumin were measured on hospital admission, immediately after delivery and 24 h later. RESULTS: Albumin and CRP levels decreased significantly in Group-II and Group-III, just after delivery. CRP increased significantly in all groups at the 24(th) hour of delivery. The effect of hemodilution on CRP and Albumin, due to volume loading, during spinal and epidural anesthesia disappeared after 24 h. CONCLUSION: We found no influence of anesthetic techniques on acute phase response except hemodilution effect. In the view of delivery types, vaginal delivery caused less stress response than section. PMID- 12554064 TI - Oxidative stress in patients with non-complicated malaria. AB - AIM: To compare oxidative stress in adults with non-complicated malaria and healthy controls. METHODOLOGY: We measured malondialdehyde (MDA), total antioxidant status (TAS), catalase, superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX). Oxidative stress was calculated based on MDA/TAS, MDA/GSH-PX and SOD/catalase indexes. RESULTS: Mean MDA in patients was 3.9 micromol/L (controls = 1.3 micromol/L). Mean TAS was 0.9 mmol/L in patients and controls. Malaria patients had less catalase activity when compared to controls (209.4 vs. 320.4 k/gr), while SOD and GSH-PX activity was higher (79.4 U/mL, 11,884.2U/L vs. 54.3 U/mL, 9,672.6 U/L). MDA/TAS index was 3.5 fold more in patients than in controls, MDA/GSH-PX and SOD/catalase indexes were increased by 6 and 2.8 fold. MDA levels and MDA/TAS index showed no differences according to malarial history, parasitaemia, Plasmodium species, parasite's stage, place of residence and drinking or smoking habits. CONCLUSIONS: During acute non-complicated P. falciparum or P. vivax malaria, we observed high oxidative stress. This resulted from lipid peroxidation rather than from a reduced TAS. We propose MDA/TAS index as a useful marker of oxidative stress during malaria infection. PMID- 12554065 TI - Absorption of three wine-related polyphenols in three different matrices by healthy subjects. AB - BACKGROUND: Despite their powerful biologic activities conducive to protection against atherosclerosis, cancer and inflammatory diseases demonstrated in vitro, there is considerable doubt whether the polyphenolic constituents present in red wine and other dietary components are effective in vivo. OBJECTIVE: We have tested the absorptive efficiency of three of these constituents (trans resveratrol, [+]-catechin and quercetin) when given orally to healthy human subjects in three different media. DESIGN: Twelve healthy males aged 25 to 45 were randomly assigned to three different groups consuming orally one of the following polyphenols: trans-resveratrol, 25 mg/70 kg; [+]-catechin 25 mg/70 kg; quercetin 10 mg/70 kg. Each polyphenol was randomly administered at 4-week intervals in three different matrices: white wine (11.5% ethanol), grape juice, and vegetable juice/homogenate. Blood was collected at zero time and at four intervals over the first four hours after consumption; urine was collected at zero time and for the following 24-h. The sums of free and conjugated polyphenols were measured in blood serum and urine by a gas-chromatographic method. RESULTS: All three polyphenols were present in serum and urine predominantly as glucuronide and sulfate conjugates, reaching peak concentrations in the former around 30-min after consumption. The free polyphenols accounted for 1.7 to 1.9% (trans-resveratrol), 1.1 to 6.5% ([+]-catechin) and 17.2 to 26.9% (quercetin) of the peak serum concentrations. The absorption of trans-resveratrol was the most efficient as judged by peak serum concentration, area-under-the curve (4 h) and urinary 24-h excretion (16-17% of dose consumed). [+]-Catechin was the poorest by these criteria (urine 24-h excretion 1.2%-3.0% of dose consumed), with quercetin being intermediate (urine 24-h excretion 2.9%-7.0% of dose consumed). Some significant matrix effects were observed for the serum polyphenol concentrations, but in the case of urine no matrix promoted significantly higher excretion than the other two. CONCLUSIONS: The absorption of these three polyphenols is broadly equivalent in aqueous and alcoholic matrices but, at peak concentrations of 10 to 40 nmol/L, is inadequate to permit circulating concentrations of 5 to 100 micromol/L consistent with in vitro biologic activity. The voluminous literature reporting powerful in vitro anticancer and antiinflammatory effects of the free polyphenols is irrelevant, given that they are absorbed as conjugates. PMID- 12554067 TI - Microbially triggered drug delivery to the colon. AB - Increasing acceptance of protein- and peptide-based drugs necessitates an investigation into the suitability of various sites for their administration. Colon is being investigated for delivery of such molecules. Colon-specific drug delivery is designed to target drug molecules specifically to this area. Development of site-specific delivery systems may exploit a specific property of the target site for drug activation/release. The gastrointestinal tract is inhabited by over 400 bacterial species, each having a specific niche in the tract. Colon, the distal part of the intestine is inhabited by a large variety of gram negative microflora. This flora produces a vast number of enzymes which are being exploited for formulation of colon-specific drug delivery systems. A number of microbially activated systems for colon-specific drug delivery are being evaluated. These include prodrugs and synthetic or natural polymer-based delivery systems. This article aims at reviewing the various microbially activated drug delivery systems for colon-specific drug delivery with specific reference to the microflora of the various segments of the gastrointestinal tract and their role in targeting drug delivery to the colon. PMID- 12554068 TI - New developments in radionuclide imaging for assessing drug delivery in man. PMID- 12554069 TI - Survival of different cell lines in alginate-agarose microcapsules. AB - Cell microencapsulation has emerged as a promising therapeutic strategy to treat a wide range of diseases. The optimisation of this technology depends on several critical issues such as the careful selection of the cell line, the controlled manufacture of microcapsules and the suitable adaptation of the construct design to the selected cell line. In this work, we studied the behavior of hybridoma cells once enclosed in solid and liquefied core alginate-agarose beads. Results show that hybridoma cells presented a better growing pattern and improved their viability and antibody production within liquefied beads. However, when these beads were evaluated with a compression resistance study, they were found to be mechanically more fragile than solid ones. To address this problem, we entrapped non-autologous cells (BHK fibroblast and C2C12 myoblast) in solid alginate agarose beads and observed that they showed an improved growing profile and prolonged their viability up to 70 days in comparison to the 15 days seen for the hybridoma cells. PMID- 12554070 TI - In vitro inhibition of simvastatin metabolism, a HMG-CoA reductase inhibitor in human and rat liver by bergamottin, a component of grapefruit juice. AB - Grapefruit juice is responsible for many drug interactions but the exact components involved in this interaction are not precisely known. Flavonoids and furocoumarin derivatives such as naringenin and bergamottin, respectively, could be involved in the inhibition of drug metabolism. The objective of this paper is to investigate in vitro the possible metabolic hepatic interaction between simvastatin (SV) and bergamottin (BG) and thus to compare its effects to those of naringenin (NRG) the aglycone form of naringin (NR) (a flavonoid present in grapefruit juice). In human and rat microsomes and in rat hepatocytes, BG was found to be a mixed type inhibitor of SV metabolism. In rat liver microsomes the K(i) value of BG (K(i)=174+/-36 microM) is higher than the K(i) value of NRG (K(i)=29+/-11 microM). However, in human liver microsomes the K(i) values are similar in BG and NRG (K(i)=34+/-5 microM and 29+/-11 microM, respectively). Moreover, it seems that there is an interspecies difference between human and rat hepatic metabolism of SV involving different isoenzymes of CYP 450. In conclusion, our study shows that BG inhibits SV metabolism. BG and NRG could therefore be applied as markers in food-drug interaction studies in order to adjust posology. PMID- 12554071 TI - Floating matrix tablets based on low density foam powder: effects of formulation and processing parameters on drug release. AB - The aim of this study was to develop and physicochemically characterize single unit, floating controlled drug delivery systems consisting of (i). polypropylene foam powder, (ii). matrix-forming polymer(s), (iii). drug, and (iv). filler (optional). The highly porous foam powder provided low density and, thus, excellent in vitro floating behavior of the tablets. All foam powder-containing tablets remained floating for at least 8 h in 0.1 N HCl at 37 degrees C. Different types of matrix-forming polymers were studied: hydroxypropyl methylcellulose (HPMC), polyacrylates, sodium alginate, corn starch, carrageenan, gum guar and gum arabic. The tablets eroded upon contact with the release medium, and the relative importance of drug diffusion, polymer swelling and tablet erosion for the resulting release patterns varied significantly with the type of matrix former. The release rate could effectively be modified by varying the "matrix-forming polymer/foam powder" ratio, the initial drug loading, the tablet geometry (radius and height), the type of matrix-forming polymer, the use of polymer blends and the addition of water-soluble or water-insoluble fillers (such as lactose or microcrystalline cellulose). The floating behavior of the low density drug delivery systems could successfully be combined with accurate control of the drug release patterns. PMID- 12554072 TI - Permeability of porcine nasal mucosa correlated with human nasal absorption. AB - The Ussing chamber diffusion system was used as a model to study the apparent permeability across porcine nasal mucosa of eight drugs and molecules with different physicochemical characteristics, namely insulin, lidocaine, nicotine, PEG 4000, propranolol, sumatriptan, melagatran and an amino diether. A weak correlation was found between the apparent permeability coefficients and the corresponding literature data on the fraction absorbed after nasal administration in humans. In the case of passively transported drugs, a closer correlation was found than for the substances where other mechanisms such as carrier-mediated transport or possible efflux were involved. Factors influencing the correlation between in vitro and in vivo data are discussed and the importance of electrophysiological control of the viability status of the excised mucosa is emphasised. Although caution has to be exercised in view of the limitations of the in vitro system, it seems to be a useful tool when evaluating different factors influencing permeability of nasal mucosa. PMID- 12554073 TI - The influence of lipid nanocapsule composition on their size distribution. AB - A formulation process, based on the inversion phase of an emulsion, was used to prepare lipid nanocapsules. Triglycerides, lecithin, salted water and hydroxy stearate of poly(ethylene glycol) were used in the preparation. The amounts of each that allowed nanocapsules to be formed described a feasibility domain within a ternary diagram. The size distribution of various nanoparticulate carriers has already been shown to influence their applications. An experimental mixture design inside the feasibility domain has been used in order to approximate, through an empirical model, the influence of the quantitative composition of nanocapsules on their size distribution. Reduced cubic polynomial equations successfully modelled the evolution of responses in terms of particle average diameters and coefficients of variation. The results were presented using an analysis of response surface showing a scale of possible particle sizes between 20 and 95 nm and a coefficient of variation between 11 and 40%. Furthermore, this technique showed that the proportion of hydrophilic surfactant had a major influence on the average diameter and the size distribution of the particles decreasing when its proportion increased. On the contrary, the coefficient of variation and the average diameter slightly increase with the proportion of triglycerides. Such a tool offers major advantages to design the formulation of particles as a function of the required size distribution. PMID- 12554074 TI - Transdermal delivery of nalbuphine and its prodrugs by electroporation. AB - The aim of this study was to assess the effects of electroporation on transdermal permeation of nalbuphine (NA) and its prodrugs. The permeation characteristics were investigated under various electrical factors and skin barriers to elucidate the mechanisms involved in transdermal delivery of NA and its prodrugs by skin electroporation. The in vitro permeation studies were performed using side-by side diffusion cells. The various electrical factors investigated were pulse voltage, pulse duration and pulse number; the different skin barriers studied were intact hairless mouse skin, stratum corneum (SC)-stripped skin, delipid skin as well as furry Wistar rat skin. The prodrugs were fully converted to parent drug after skin permeation. Application of electroporation significantly enhanced transdermal permeation of NA and its prodrugs. The enhancement ratios were highest for NA and the four prodrugs showed the similar permeability after electroporation. The permeation amounts of NA and its prodrugs may be increased by application of higher pulse voltage, pulse duration as well as pulse number. Various kinetics and mechanisms were observed for the permeation of the hydrophilic NA and lipophilic nalbuphine enanthate through different skin barriers by applying electroporation. This study demonstrated that electroporation may enhance and control transdermal permeation of NA and its prodrugs. The results also indicated that the physicochemical properties of prodrug had significant effects on kinetics as well as mechanisms of transdermal permeation by electroporation. PMID- 12554075 TI - Transdermal delivery of zidovudine: effect of vehicles on permeation across rat skin and their mechanism of action. AB - The purpose of this study was to investigate the effects of various solvent systems containing water, ethanol, propylene glycol (PG), and their binary combinations on the ex vivo permeation of zidovudine (AZT) across Sprague-Dawley rat skin using Franz diffusion cells at 37 degrees C. Further, saturation solubility and epidermis/vehicle partition coefficient of AZT in the solvent systems, and their effect on percentage hydration of epidermis using thermogravimetric analysis were determined to understand the mechanisms by which these solvent systems change drug permeability properties. All binary combinations of PG, ethanol and water significantly increased saturation solubility of AZT. Maximum AZT flux was observed with 66.6% ethanol among ethanol water solvents, with 33.3% PG in PG-water solvents and with 100% ethanol among PG ethanol combinations. PG-water and PG-ethanol solvents neither reduced the lag time nor increased AZT flux across rat skin. In addition, high concentrations of PG in both water and ethanol reduced steady state flux of AZT. Further, thermogravimetric studies revealed that solvents containing high PG concentrations dehydrate epidermis. Among all the solvent combinations, highest flux and short lag time were achieved with ethanol at 66.6% in water and hence is a suitable vehicle for transdermal delivery of AZT. PMID- 12554076 TI - Effects of cholesterol on dye leakage induced by multidrug-resistance modulators from anionic liposomes. AB - Multidrug-resistance (MDR) in cancer cells is often associated with marked changes in the membrane cholesterol levels. To assess the cholesterol-dependence of MDR modulator efficiency in terms of the drug-membrane interactions, the ability of 5 MDR-modulators to induce the leakage of Sulphan blue through anionic liposomes was quantified at various mole fractions x(chol) of cholesterol (0 0.42). Depending on the electric charge of the drug, cholesterol modified to a large extent either the permeation dose inducing 50% dye leakage (PD(50)) or the co-operativity (h) of the permeation process. The PD(50) of Triton X-100 (non ionic) and that of diltiazem and verapamil (mono-basic amines) varied only slightly (0.3 mM) with the cholesterol level, whereas the co-operativity increased by 1.9-2.7. On the reverse, the PD(50) of a thioacridine derivative and mepacrine (di-basic amines) increased by 4.8-7.5 mM in the cholesterol range investigated, whereas the co-operativity (h) increased slightly (0.2-0.7). In the permeation process, the rate-limiting character of the electric charge (z) of the drug is likely to be strengthened by high cholesterol levels. The results provide evidence that in resistant tumours exhibiting high cholesterol levels, the MDR might be reversed by favourable drug-membrane interactions if the modulators are designed in the form of highly lipophilic mono-basic drugs that counteract the effects of cholesterol on the membrane dipolar potential and membrane fluidity. PMID- 12554077 TI - Mucoadhesive and cohesive properties of poly(acrylic acid)-cysteine conjugates with regard to their molecular mass. AB - The objective of this study was to evaluate the influence of the molecular mass and accordingly the polymer chain length on mucoadhesion and cohesion of thiolated polymers. Linear poly(acrylic acid)-cysteine (PAA-Cys) conjugates of 2 , 45-, 250- and 450 kDa (PAA(2)-Cys, PAA(45)-Cys, PAA(250)-Cys and PAA(450)-Cys) and polycarbophil-cysteine (PCP-Cys, 750-3000 kDa), all displaying on average 404.1+/-65.5 microMol thiol groups per gram polymer were compressed into tablets to perform disintegration tests, mucoadhesion studies and viscosity measurements. Moreover, the influence of free unbound cysteine on mucoadhesion was evaluated. Disintegration tests showed a stability of the tablets as following: PAA(2) Cys3Galpbeta1-->6(Galfbeta1-->4)GlcNAc, is expressed on the sialoglycoprotein of Trypanosoma cruzi Dm28c. PMID- 12554089 TI - Effect of N-terminal truncation of Plasmodium falciparum dihydrofolate reductase on dihydrofolate reductase and thymidylate synthase activity. PMID- 12554090 TI - Structural characterization of the N-linked glycans from Taenia solium metacestodes. PMID- 12554091 TI - The Plasmodium falciparum fatty acyl-CoA synthetase family (PfACS) and differential stage-specific expression in infected erythrocytes. PMID- 12554092 TI - Cloning and characterization of a gene encoding a putative protein associated with U3 small nucleolar ribonucleoprotein in Trypanosoma cruzi. PMID- 12554093 TI - Phylogenetic analysis of Blastocystis isolates from different hosts based on the comparison of small-subunit rRNA gene sequences. PMID- 12554094 TI - The lymphatics revisited: new perspectives from the hyaluronan receptor LYVE-1. AB - The lymphatic vasculature forms a second circulatory system that drains extracellular fluid from the tissues and provides an exclusive environment in which immune cells can encounter and respond to foreign antigen. Although equally important for the functioning of both the cardiovascular and immune systems, there is only a rudimentary knowledge of the molecular biology of lymphatics, owing to longstanding difficulties in identifying and isolating lymphatic endothelial cells. Recently, however, a number of interesting molecules have been identified that may be exploited as markers for lymphatic endothelium, including the hyaluronan receptor LYVE-1, the subject of this review. This article considers the biology of LYVE-1 and outlines how recent research into this molecule is providing new insights into the nature of lymphatics and their role in tumor metastasis. PMID- 12554095 TI - Exosite-dependent regulation of the protein C anticoagulant pathway. AB - Activated protein C (APC) is a vitamin K-dependent anticoagulant serine protease in plasma that downregulates the coagulation cascade by degrading cofactors Va and VIIIa by limited proteolysis. In addition to its anticoagulant function, APC also exhibits potent profibrinolytic and anti-inflammatory properties. The proteolytic activity of APC in plasma is slowly inhibited by three serpins: protein C inhibitor, plasminogen activator inhibitor-1, and alpha(1)-antitrypsin. Recent structural and mutagenesis data have indicated that basic residues of three exposed surface loops known as the 39-loop (Lys(37)-Lys(39)), 60-loop (Lys(62), Lys(63)), and 70-80-loop (Arg(74), Arg(75), and Lys(78)) (chymotrypsin numbering) constitute an anion-binding exosite in APC that interacts with these macromolecular substrates and inhibitors. Moreover, this exosite plays a critical role in the thrombomodulin-dependent activation of the zymogen protein C by thrombin. This article briefly reviews how the binding of physiological protein and polysaccharide cofactors on this exosite modulates the protein C anticoagulant pathway in plasma. PMID- 12554096 TI - The role of modulatory calcineurin-interacting proteins in calcineurin signaling. AB - Modulatory calcineurin-interacting proteins (MCIPs), also known as the Down syndrome critical region 1 (DSCR1) and DSCR1-like proteins, are a recently described family of small, structurally related proteins that are preferentially expressed in heart, skeletal muscle, and brain. MCIP proteins can bind to and inhibit calcineurin, a calcium/calmodulin-regulated serine/threonine protein phosphatase that is activated during cardiac hypertrophy and failure. Transcription of the mammalian MCIP1 gene is induced by calcineurin, suggesting that it functions as an endogenous feedback regulator of calcineurin signal transduction. Forced expression of human MCIP1 protein in the hearts of transgenic mice attenuates the hypertrophic response to a broad range of stimuli. This review summarizes work from a number of laboratories on the structure, regulation, and function of MCIP proteins. PMID- 12554097 TI - Critical role for conversion of glu-plasminogen to Lys-plasminogen for optimal stimulation of plasminogen activation on cell surfaces. AB - When Glu-plasminogen, the native circulating form of the zymogen, is bound to cell surfaces, its activation is markedly enhanced compared with the reaction in solution. This results in localization of the broad-spectrum proteolytic activity of plasmin on cell surfaces. The cell-associated plasmin plays a key role in fibrinolysis, cell migration, and prohormone processing. It is well established that the localization of plasminogen and plasminogen activators on cell surfaces promotes the enhanced plasminogen activation on the cell surface. The focus of this article is to review recent studies demonstrating that the conversion of Glu plasminogen to the more readily activated Lys-plasminogen derivative is necessary for optimal stimulation of plasminogen activation on the cell surface, and that the interaction of Glu-plasminogen with cells serves to increase processing of Glu-plasminogen to Lys-plasminogen, thereby enhancing plasminogen activation on the cell surface. PMID- 12554098 TI - Dilated cardiomyopathy: a disease of the intercalated disc? AB - The contractile tissue of the heart is composed of individual cells, making specific cell-cell contacts necessary to ensure mechanical and electrochemical coupling during beating. These contact sites, termed the intercalated discs, have gained increased attention recently due to their potential involvement in cardiac disease. This article discusses how the intercalated discs are assembled during heart development and how they are affected in cardiomyopathy, with particular emphasis on dilated cardiomyopathy. A model is proposed to relate the alterations that are seen at a molecular level with changes in function observed in that kind of cardiac disease. PMID- 12554099 TI - Structure-function relationships in factor IX and factor IXa. AB - Factor IX (FIX) consists of an N-terminal gamma-carboxyglutamic acid (Gla) domain followed by two epidermal growth factor (EGF)-like domains, and the C-terminal serine protease domain. During physiologic coagulation, one of the activators of FIX is the FVIIa/tissue factor (TF) complex. In this reaction, the Gla and EGF1 domains of FIX are thought to interact with TF. The FIXa that is generated then combines with FVIIIa on the platelet surface to activate FX in the coagulation cascade. In this assembly, the protease domain and possibly the EGF2 domain of FIXa are thought to provide the primary specificity in binding to FVIIIa. Disruption of the interaction of FIX/FX with TF and of the FIXa:FVIIIa interface may provide a pharmacologic target as an alternative strategy for the development of antithrombotic agents. PMID- 12554100 TI - Epidemiology of Norwalk-like virus infections in cattle in The Netherlands. AB - "Norwalk-like viruses" (NLVs) are the most common cause of acute non-bacterial gastroenteritis in humans. Cattle may be a reservoir of NLVs although never bovine NLVs have been found in humans. To gain more insight into the epidemiology of NLV, infections in cattle in The Netherlands were studied. Individual faecal samples from a large dairy herd and 243 pooled samples from veal calf farms were analysed for NLV by RT-PCR. Calves under 3 months of age in the dairy herd were sampled three to five times with 3-week intervals, whereas dairy cattle were sampled twice with a 2-month interval. In 31.6% (77/243) of the veal calf farm samples and in 4.2% (13/312) of the individual dairy cattle samples NLV was detected. The mean age of virus positive dairy cattle was 2.5 months. The highest numbers of NLV positive veal calf farms in The Netherlands were found in the regions with the highest number of veal calf farms. NLV infected veal calf farms were detected in every month throughout the study period. Cattle appeared to be hosts of NLVs, and virus shedding was weakly associated with diarrhoea. Complete ORF2 sequences were obtained from two calf NLVs and phylogenetic analyses suggested that these strains belong to a distinct cluster (GGIII/2) in between GGI and GGII NLVs of humans. Overall, genetic variation between strains as determined by sequence analysis of the P1/P2 capsid region was limited to 14.6%. Our data shows that NLV is endemic in the cattle population in The Netherlands and genetically distinct from NLVs in humans. PMID- 12554101 TI - Determination of the sequence of the complete open reading frame and the 5'NTR of the Paderborn isolate of classical swine fever virus. AB - The classical swine fever (CSF) epidemic in the Netherlands in 1997-1998 lasted 14 months, during which 429 infected and 1300 at risk herds were culled, at an estimated economical cost of 2 billion US dollars. Despite the overwhelming scale of the epizootic, the CSF virus (CSFV) strain causing the outbreak has remained largely uncharacterized. The Dutch epizootic is epidemiologically linked to a small CSF outbreak in 1997, in Paderborn in Germany. E2 and partial 5' NTR sequencing has shown that the index Paderborn isolate, and several Dutch isolates taken during the 1997-1998 epizootic, are virtually identical, confirming that the Paderborn isolate triggered the Dutch outbreak, and furthermore showing that this single isolate was stable throughout the whole Dutch outbreak (the above reviewed in [C. Terpstra, A. J. de Smit, Veterinary Microbiol. 77 (2000) 3-15]). We determined the nucleotide sequence of the 5' NTR (by 5' RACE) and the complete open reading frame of the Paderborn isolate (GenBank AY072924). Our sequence was identical to previously published partial 5'NTR and E2 sequences for the index Paderborn 1997 and Dutch 1997 (Venhorst) isolates, confirming the identity of the virus we sequenced. Phylogenetic analysis based on the complete open reading frame showed that Paderborn is genetically very different from common European laboratory reference strains. Neutralization studies showed that Paderborn is also antigenically very different from common laboratory strains such as Alfort 187. Paderborn is the only recent European CSFV field isolate for which a complete sequence is available, and given Paderborns genetic and antigenic uniqueness, the Paderborn sequence may have practical use for diagnostic and vaccine antigen development. PMID- 12554102 TI - Staphylococcus aureus isolates from dairy cows and humans on a farm differ in coagulase genotype. AB - Staphylococcus aureus is a frequent cause of animal and human infections. The aim of the present study was to test diversity of the populations of S. aureus colonising cattle and humans sharing an infected environment. Eighty-six S. aureus isolates obtained from dairy cows, from people coming into contact with dairy cows on the farm and the other farm personnel were characterized by restriction fragment length polymorphism of the coagulase gene. Molecular analyses identified ten polymorphism types with prevalent presentation of type II in isolates from cow's milk and type IV in isolates from people coming into contact with dairy cows on the farm (the cattlemen) and the other farm personnel. Seven further genotypes were identified among the isolates from the cattlemen. The results indicate that the strains dominating in human population did not equate to the causative agents of bovine mastitis. PMID- 12554103 TI - Serotypes and Shiga toxin genotypes among Escherichia coli isolated from animals and food in Argentina and Brazil. AB - Shiga toxin (Stx)-producing Escherichia coli (STEC) strains isolated from animals and food in Argentina (n=44) and Brazil (n=20) were examined and compared in regard to their phenotypic and genotypic characteristics to evaluate their pathogenic potential. The clonal relatedness of STEC O157 isolates (n=22) was established by phage typing (PT) and pulsed-field gel electrophoresis (PFGE). All O157 strains studied carried eae and enterohemorrhagic E. coli (EHEC)-hly sequences. In Argentina, these strains occurred both in cattle and meat, and 50% of them carried stx2/stx2vh-a genes, whereas in Brazil the O157 strains were isolated from animals, and most harbored the stx2vh-a sequence. At least 13 different O:H serotypes were identified among the non-O157 strains studied, with serotype O113:H21 being found in both countries. All but one non-O157 strains did not carry eae gene, but EHEC-hlyA gene was found in 85.7% of them, and the stx2 genotype was also more prevalent in Argentina than in Brazil (P<0.01), where stx1 alone or in association was most common (68.8%). One STEC strain isolated from a calf in Brazil harbored the new variant referred to as stx2-NV206. PFGE analysis showed that STEC O157 strains were grouped in four clusters. One Brazilian strain was considered possibly related (> or =80%) to Argentinean strains of cluster I. Differences in the pathogenic potential, especially in regard to serotypes and stx genotypes, were observed among the STEC strains recovered from animals and food in both countries. PMID- 12554104 TI - Discrimination among Listeria monocytogenes isolates using a mixed genome DNA microarray. AB - Listeria monocytogenes can cause serious illness in humans, usually following the ingestion of contaminated food. Epidemiologic investigation requires identification of specific isolates, usually done by a combination of serotyping and subtyping using pulsed-field gel electrophoresis (PFGE). DNA microarrays provide a new format to resolve genetic differences among isolates and, unlike PFGE, to identify specific genes associated with the infecting pathogen. A 585 probe, mixed genome microarray was constructed and 24 strains of L. monocytogenes were hybridized to the array. Microarray analysis allowed discrimination among L. monocytogenes isolates within a serotype and obtained from similar geographic and epidemiologic sources. Importantly, the microarray results preserved previously described phylogenetic relationships between major serogroups and, in a limited comparison, agreed with PFGE subtypes. The association of individual probes with isolates allowed identification of specific genes. Sequencing of 10 polymorphic probes identified nine matches with previously described bacterial genes including several suspected virulence factors. These results demonstrate that mixed genomic microarrays are useful for differentiating among closely related L. monocytogenes isolates and identifying genetic markers that can be used in epidemiologic and possibly pathogenesis studies. PMID- 12554105 TI - An RTX operon in hemolytic Moraxella bovis is absent from nonhemolytic strains. AB - Pathogenic isolates of Moraxella bovis express a calcium-dependent transmembrane pore forming cytotoxin that is an RTX toxin encoded by mbxA. The DNA flanking mbxA was cloned and sequenced to determine if M. bovis contained a classical RTX operon. Open reading frames (ORFs) with deduced amino acid sequence homology to putative activation (RTX C) and transport (RTX B and D) proteins were identified and have been designated MbxC, MbxB, and MbxD, respectively. Thus, hemolytic M. bovis contains a typical RTX operon comprised of four genes arranged (5'-3') mbxCABD. In addition, the deduced amino acid sequences of DNA flanking mbxCABD revealed ORFs with amino acid sequence similarity to transposases (5'). At the 3' end of the mbx gene cluster, an ORF with homology to bacterial tolC genes was identified. Thus, as with the cya RTX operon of Bordetella pertussis, M. bovis appears to have a secretion accessory protein linked to RTX genes. Analysis of genomic DNA isolated from 5 nonhemolytic M. bovis strains by PCR and Southern blotting revealed the absence of mbxCABD. These strains did, however, amplify with primers specific for the 5' region flanking mbxC. M. bovis harbors a classical RTX operon that is absent in nonhemolytic strains. PMID- 12554106 TI - Comparison of Salmonella enterica serovar Abortusequi isolates of equine origin by pulsed-field gel electrophoresis and fluorescent amplified-fragment length polymorphism fingerprinting. AB - Equine paratyphoid is caused by Salmonella enterica serovar Abortusequi, and manifests mainly as abortion in the mare. We compared S. Abortusequi strains isolated in Japan and other countries using pulsed-field gel electrophoresis (PFGE) and fluorescent amplified-fragment length polymorphism (FAFLP) analysis. PFGE analysis of S. Abortusequi strains gave 21-27 fragments ranging in size from 33 to 602kb. Although two PFGE profiles were observed among the 20 S. Abortusequi isolates in Japan, the restriction fragments originating from the chromosome were common between the two profiles. The similarity index of the two profiles was 90.9%, while those between Japanese and five other S. Abortusequi strains were 29.8-37.5%. On the other hand, FAFLP analysis of S. Abortusequi strains generated 64-67 amplified fragments ranging in size from 100 to 400bp. One polymorphic fragment was observed among the 20 S. Abortusequi isolates in Japan. These data indicate the close relation of this agent in Japan. S. Abortusequi strains sharing a common ancestry might have been conserved in Japan. PMID- 12554107 TI - The influence of functional forces on the biomechanics of implant-supported prostheses--a review. AB - OBJECTIVES: To evaluate published evidence related to the influence of functional forces on the biomechanics of implant-supported prostheses. DATA AND SOURCES: The literature was searched for original research articles relating control of loads on dental implants, effects of early and late occlusal loads, the influence of bone quality, prosthesis type, prosthesis material, number of supporting implants, and engineering techniques employed for evaluating mechanical and biomechanical behavior of implants using MEDLINE and manual tracing of references cited in key papers otherwise not elicited. STUDY SELECTION: Current literature on implant biomechanics as main focus and pertinent to key aspects of the review. CONCLUSIONS: The outcome of implant treatment is often maximized when implants are placed in dense bone, number of supporting implants are increased, implant placement configuration reduces the effects of bending moments, and when a fixed prosthesis is delivered to the patient. PMID- 12554108 TI - Stresses at the cervical lesion of maxillary premolar--a finite element investigation. AB - OBJECTIVES: The objective of this study was to use a three-dimensional (3-D) finite element model to investigate normal stress distribution to substantiate the tooth flexure mechanism. The study also compared the changes in the stresses by different occlusal loading sites and directions. METHODS: The 3-D finite element analysis was used. A maxillary premolar was selected to construct the simulation model. The model was constructed step-by-step for convergence and validity. Seven load conditions for various load sites and different directions were simulated to the model. RESULTS: The maximal principal stress and minimal principal stress distributions developed within the structures of seven load conditions were output and their stress distributions on z-plane at the vertical midline were shown. The peak tensile stress of the cervical area for various load conditions were compared and listed. CONCLUSIONS: This study has shown that the presence of tensile stresses in the cervical region of a maxillary premolar by various loading sites and different directions. The results coincided with the stress-induced theory, hence sustaining it. The relationship of the affected factors of leverage to the development of cervical abfraction lesions, was explored. PMID- 12554109 TI - Pulpal space pressure and temperature changes from Nd:YAG laser irradiation of dentin. AB - OBJECTIVES: To investigate pulpal space pressure and temperature after application of Nd:YAG laser, and high-speed diamond bur on dentin surface. METHODS: One and 3 W Nd:YAG laser and high-speed diamond bur were used to remove dentine from twenty extracted premolars. The pulp chambers were monitored for pressure and temperature changes with a pressure transducer and thermocouple, respectively. RESULTS: Regardless of the remaining dentin thickness (RDT), laser irradiation and high-speed diamond bur use generated an increase in pulpal space pressure and temperature (ANOVA and Fisher's LSD tests, P < 0.001). Pressure and temperature increased with an increase in laser power. Three-Watt laser irradiation caused greater changes than 1 W (1.75 kPa and 1.31 degrees C, 0.53 kPa and 0.34 degrees C, respectively). Both pulpal space pressure (P < 0.001) and temperature (P < 0.005) increased as the RDT decreased. CONCLUSIONS: Laser irradiation and the use of a high-speed diamond bur generated an increase in pulpal space pressure and temperature. Pulpal space pressure and temperature increased with an increase in energy density of laser and a decrease in RDT. PMID- 12554110 TI - Hierarchy of variables correlated to odontoblast-like cell numbers following pulp capping. AB - OBJECTIVES: Following tooth pulp exposure, pulpal repair is accomplished by dentine bridge secretion by odontoblast-like cells. However, little information is available about the hierarchy of variables, which influence odontoblast-like cell numbers. The purpose of this study was to examine correlations between pulp capping events and odontoblast-like cell numbers. METHODS: Two hundred and fifty standardised pulp exposed cavities were prepared in non-human primate teeth according to ISO usage guidelines. Exposed pulps were capped with Calcium hydroxide [Ca(OH)(2)], and multi-step and self-etching primer composite resins. Teeth were collected from 3 to 60-days to observe pulp reactions. Statistical analysis was evaluated using analysis of variance. RESULTS: The hierarchy of variables correlated to odontoblast-like cells were the dentine bridge area (P = 0.0001), time since pulp exposure (P = 0.0001), odontoblast numbers opposite the exposure site (P = 0.0002), and pulp capping materials (P = 0.0313). Other pulp capping variables were found to be less likely to be correlated with odontoblast like cell numbers. CONCLUSIONS: The area of dentine bridge formation is directly related to the numbers of odontoblast-like cells, cell activity is time dependent, and the cell numbers are much lower than original odontoblast cells. The time-lag between the appearance of odontoblast-like cells at the site of pulp exposure, and the limited numbers of these cells, explain why pulpal repair is difficult to achieve successfully following pulp exposure. PMID- 12554111 TI - In vitro assessment of incipient approximal carious lesions using computer assisted densitometric image analysis. AB - OBJECTIVES: To investigate the use of computer-assisted densitometric image analysis (CADIA) to detect incipient approximal carious lesions in vitro and to compare areas of demineralization and mechanically induced lesions by radiographic visual inspection. METHODS: Thirty-six extracted non-carious human molars were demineralized in a defined approximal area of interest (AOI) using a gel system (pH 4.8) applied over 15 weeks. Standardized digital radiographs were taken before and after creation of caries-like lesions and grayscale values were measured within the AOI. For another group of eight teeth, artificial defects were mechanically induced by a diamond bur and densitometric analysis was performed. Statistical significance was tested using the Wilcoxon test and set at p < or = 0.05. In addition, radiographs were interpreted by visual examination by six experienced dentists. The sensitivity and specificity for both study groups were determined in comparison to a baseline group of untreated teeth. Polarized light microscopy and SEM analysis were used to evaluate changes in the subsurface mineralization pattern and surface micromorphology. RESULTS: Densitometric comparison of images from identical regions in control teeth revealed no significant grayscale value differences within the AOI, whereas a significant decrease (p < or = 0.05) was observed in the demineralized group. Mechanically induced lesions also showed a significant densitometric decrease (p < or = 0.05). The sensitivity of visual inspection of radiographs of demineralized areas was 0%. In contrast, the sensitivity for the detection of a mechanical defect of 0.1mm, which was the same subsurface extension as the histologically determined demineralization, increased to 52%. Specificity was 100% for all groups. CONCLUSIONS: CADIA is able to detect very small changes in mineralization of incipient carious approximal lesions in vitro. The findings presented in this investigation may improve the subtraction method used for diagnosis and monitoring of approximal caries, as well as prophylactic measures. PMID- 12554112 TI - The pH and titratable acidity of a range of diluting drinks and their potential effect on dental erosion. AB - OBJECTIVES: To investigate, in vitro, the potential dental effects of diluting juices by measuring their acidity levels and to examine the effect dilution has on the potential erosive properties of the drinks. METHODS: Four commonly available diluting drinks were assessed for both pH and titratable acidity at a variety of dilutions, reflecting the range of concentrations over which such drinks may be consumed. Predetermined dilutions of citric acid and hydrochloric acid, with similar pH values to those of the drinks, were used as positive control acidic solutions. The pH measurements throughout the study were made using a pH electrode connected to an Orion EA940 IonAnalyser. RESULTS: All four drinks demonstrated a high degree of resistance to a rise in pH, indicating high intrinsic buffering capacity. The measured pH changed very little with increasing dilution ratio when compared to the citric and hydrochloric acid controls, even when considered extremely dilute. In contrast, the titratable acidity of each of the drinks reduced proportionally with increasing dilution, thereby reducing considerably its erosive potential. CONCLUSIONS: While dilution had very little effect on the measured pH values, the titratable acidity reduced as the drink became more dilute. Since there appears to be a direct relationship between dilution and titratable acidity, the erosive potential of diluting juices may be reduced substantially by the addition of water. It remains unlikely, however, that the drinks would ever be consumed at a 'safe' level of dilution due to diminished taste and colour. PMID- 12554113 TI - The effects of the combination of chlorhexidine/thymol- and fluoride-containing varnishes on the severity of root caries lesions in frail institutionalised elderly people. AB - OBJECTIVES: To compare the clinical effects of a fluoride-containing varnish (Fluor-Protector) in combination with a chlorhexidine-containing varnish (Cervitec) on existing root caries lesions in a group of frail elderly subjects. METHODS: A randomised double blind longitudinal study was utilised. Subjects (n = 102) were randomly allocated to a Test or Placebo group. All leathery and soft root caries lesions in all subjects were coated with Fluor-Protector while the lesions in the Test group were also coated with Cervitec and the lesions in the Placebo group were coated with a Placebo varnish. Treatments were repeated five times in a 12-month period. Clinical parameters associated with root caries, measurements of individual lesions and salivary levels of caries associated bacteria were made at intervals. RESULTS: The clinical severity of the lesions in the Test group did not change significantly during the 12-month study period. In the Placebo group the mean lesion width and lesion height and length of exposed root increased significantly and the lesions were significantly closer to the gingival margin. There were no significant changes in the salivary levels of caries-associated microorganisms after 12 months although, in both groups, there was initially a significant reduction in the salivary levels of mutans streptococci. CONCLUSIONS: The combination of Fluor-Protector and Cervitec is a useful, simple, quick and non-invasive method for the control and management of existing root caries lesions. The procedure could be performed by a dental hygienist and may be usefully applied in other high-risk groups including persons with Parkinson's disease, debilitating neuromuscular conditions and dry mouth from whatever cause. PMID- 12554114 TI - Effects of nitrite and nitrate on the growth and acidogenicity of Streptococcus mutans. AB - It is hypothesised that exogenous nitrite acidified by metabolic products of acidogenic bacteria in the mouth will be converted to products which inhibit growth of the bacteria in question which contribute to dental caries. OBJECTIVES: The aims of this study were (1) to test the activity of both sodium nitrate and sodium nitrite at differing concentrations on the ability of Streptococcus mutans to lower the pH of its surroundings and hence (2) to determine whether either nitrate or nitrite might be bactericidal or bacteriostatic against S. mutans. METHODS: S. mutans NCTC 10449(T) was cultured in a liquid medium to which either sodium nitrate or sodium nitrite was added to a final concentration of 0.0, 0.2, 2.0, 20 or 200 mM, of which the first acted as a test substance negative control. After 24 h, the cultures were streaked onto agar to test for growth and the remaining culture used for pH measurement. The Mann-Whitney U-Test was used for statistical comparison of pH values. RESULTS: Nitrite at concentrations of 20 and 200 mM had a highly significant inhibitory effect (p < 0.001) on the ability of S. mutans NCTC 10449(T) to lower pH. Moreover, bacteria that had been subjected to these levels of nitrite were unable to recover on solid medium. Nitrate had no such effect on either the growth of the bacteria or on their ability to lower pH. CONCLUSIONS: It is concluded that nitrite, at final concentrations of either 20 or 200 mM, is both bactericidal and anti-acidogenic with respect to S. mutans, while lower concentrations of nitrite and all concentrations of nitrate are ineffective. Nitrite might be worthy of consideration as a mouth-rinse constituent. PMID- 12554115 TI - Polymerization contraction stress in dual-cure cements and its effect on interfacial integrity of bonded inlays. AB - OBJECTIVES: To compare the polymerization contraction stress of resin cements in dual-cure and self-cure modes, and verify its effect on microleakage and gap formation in class I cavities restored with porcelain inlays. METHODS: Dual-cure cements (Calibra, Choice and RelyX ARC) were monitored for contraction stress during 30 min in the tensilometer, in the presence or absence of photo activation. Microleakage was evaluated in porcelain inlays bonded to cavities prepared in bovine incisors, combining the same adhesive (One-Step) with each of the cements in both activation modes. Epoxy replicas of the sectioned specimens were analyzed in the SEM to determine the percentage of discontinuous margins. RESULTS: The stress developed in dual-cure mode was higher than in self-cure mode for all cements. In self-cure mode, no significant difference in stress was found among materials. Choice and RelyX ARC showed higher microleakage in dual-cure mode than in self-cure mode. Calibra did not show a significant difference in microleakage between activation modes. Contraction gap results showed wide variations, and no significant differences were detected among the groups. Gaps occurred more often in dentin, with an overall average of 22 +/- 17.5%. CONCLUSIONS: The higher stress generated in dual-cure cements subjected to photo activation may lead to higher marginal leakage. In the confinement condition represented by the class I inlay, even the lower contraction stress of the cements in self-cure mode had enough magnitude to disrupt the bonding to dentin. PMID- 12554116 TI - Erosion and attrition of human enamel in vitro part I: interaction effects. AB - OBJECTIVE: The aim of the study in vitro was to measure the interplay of attrition and erosion of human enamel under several different pH conditions. MATERIALS AND METHODS: Cusp and smooth surface enamel specimens were prepared from unerupted human third molar teeth. Cusp and smooth surface specimens opposed each other in a wear machine under a load of 600 g. The machine simulated tooth grinding for 30 min (2280 wear cycles). Six groups of 10 specimens were worn in the following environmental conditions: saline, citric acid at pH values of 3.2, 5.5 or 7.0 and two cycling groups (pH 3.2/saline or saline/pH 3.2). Additionally, 10 smooth surface specimens were exposed to the same fluids without attrition. Tissue loss on smooth surface specimens was determined by profilometry. The worn specimen surfaces were studied by SEM. RESULTS: Attrition depth in saline and pH 7.0 citric acid was 25.5 microm. At pH 3.2 and 5.5 attrition depths were 6.0 and 4.3 microm, respectively. Cycling in saline/citric acid and vice versa produced attrition depths of 9.2 and 7.9 microm, respectively. In the erosion only specimens lesion depths were 3.0, 1.2 and 0 microm at pH 3.2, 5.5 and 7.0 and saline, respectively. Attrition specimens at pH 7.0 and in saline showed very rough surfaces. At low pH enamel surfaces appeared flat and smooth. CONCLUSIONS: Enamel wear in neutral conditions is significantly higher than in acidic conditions (p < 0.05), apparently due to a smoothing effect of erosion on contacting surfaces. PMID- 12554117 TI - Erosion and attrition of human enamel in vitro part II: influence of time and loading. AB - OBJECTIVES: To determine the influence of load and time on enamel wear by attrition under neutral and acid environments in vitro. METHODS: Groups of 10 enamel cusp and smooth surface specimens obtained from unerupted third molars were opposed in a wear machine under loads of 200, 400 and 600 g in citric acid at pH 3.2 and in physiological saline at 35 degrees C for 30 min (2280 wear cycles). The experiment was repeated with a load of 600 g for 10 and 20 min accompanied by 10 taped specimens to measure erosion only. Erosion and attrition of smooth surface specimens was determined by profilometry. RESULTS: At a load of 600 g attrition in saline increased with time from 7.9 microm (SD 3.7 microm) at 10 min, 10.2 microm (SD 4.8 microm) at 20-25.5 microm (SD 9.1 microm) after 30 min and combined attrition/erosion in citric acid from 1.5 microm (SD 0.4 microm) to 2.5 microm (SD 0.9 microm) to 6.0 microm (SD 3.7 microm). At a fixed time of 30 min, attrition in saline was 5.2 microm (SD 3.6 microm) at 200 g and 8.1 microm (SD 1.5 microm) at 400 g and combined attrition/erosion in citric acid 2.8 microm (SD 1.2 microm) and 3.6 microm (SD 0.7 microm). Comparison between both environments revealed significant differences at fixed load at all time points (p < 0.01) and at fixed time between 400 and 600 g (p < 0.01). CONCLUSIONS: Load and time significantly influence enamel wear both in acid and neutral conditions but the effects were not linear. PMID- 12554119 TI - A method for the detection and quantification of bacteria in human carious dentine using fluorescent in situ hybridisation. AB - OBJECTIVES: Previous studies have evaluated bacterial numbers in carious dentine using conventional culturing methods, capable of detecting only a proportion of the total bacteria present within lesions. The aim of this study was to detect and enumerate the total bacterial population present in carious human dentine by means of fluorescent in situ hybridisation. METHOD: Five freshly extracted carious molars were sequentially hand excavated under sterile conditions through four levels in the lesions. Replicate samples were probed with a rhodamine tagged, 16S rRNA-directed probe (EUB338), specific for the bacterial domain. Two of the five original samples were examined using fluorescence microscopy and by using a systematic visual counting strategy; direct enumeration of the bacterial population in carious dentine was performed. RESULTS: In the superficial, middle and deep/excavation front zones, a mean of 7.34 x 10(6) (standard error of the mean, SEM +/- 0.44), 5.23 x 10(6) (SEM +/- 0.18), and 1.69 x 10(6) (SEM +/- 0.15) total bacteria/mg dentine were found, respectively. In the advancing front zone (beyond the conventional clinical excavation boundary) a mean of 0.34 x 10(6) (SEM +/- 0.05) total bacteria/mg dentine was recorded. CONCLUSION: A bacterial enumeration strategy was developed and detected greater numbers of bacteria through the depth of carious lesions that had been reported previously. The technique could be further developed using species-specific probes to determine the distribution, abundance and viability of all bacteria in carious dentine. This new information in turn will lead to a better understanding of the pathological process of caries and ultimately, its clinical treatment. PMID- 12554118 TI - Toxicokinetic of HEMA in guinea pigs. AB - OBJECTIVE: Unconverted 2-hydroxyethylmethacrylate (HEMA) can be released from dental resin materials and can enter the body in humans. In the present study the uptake, distribution and excretion of 14C-HEMA applied via different routes were examined in vivo in guinea pigs. METHODS: HEMA (0.02 mmol/kg bw labelled with a tracer dose 14C-HEMA 0.3 Bq/g bw) was administered by gastric tube or by subcutaneous injection. Urine, feces, and exhaled carbon dioxide were collected for 24 h after administration. Guinea pigs were killed 24 h after the beginning of the experiment and various organs removed and 14C radioactivity measured. RESULTS: Low fecal 14C levels (about 2% of the dose) and urinary levels of about 15% after 24 h were noted with either route of administration. Direct measurement of exhaled CO(2) showed that about 70% of the dose left the body via the lungs. Two pathways for the metabolism of 14C-HEMA can be described. It is likely that 14C-pyruvate is formed in vivo resulting in the formation of toxic 14C-HEMA intermediates. 14C-HEMA was taken up rapidly from the stomach and small intestine after gastric administration and was widely distributed in the body following administration by each of the routes. CONCLUSIONS: Clearance from most tissues following gastric and intradermal administration was essentially complete within one day. The peak HEMA levels in all tissues examined after 24 h were at least onemillion-fold less than known toxic levels. PMID- 12554120 TI - The effect of unstimulated and stimulated whole saliva on extrinsic staining in vitro--a developmental method. AB - OBJECTIVES: To detect any differences in the propensity of unstimulated and stimulated, individual whole saliva to cause in vitro staining by chlorhexidine and tea. METHODS: Unstimulated and stimulated human saliva was collected on a daily basis and used to coat optically clear acrylic specimens. Specimens were subjected to an established chlorhexidine/tea staining model in vitro shown to correlate well with in vivo staining, and cycles repeated until an optical density of <2 was reached. RESULTS: Stain development increased incrementally with increasing cycles. Overall differences in chlorhexidine/tea staining were noted both between subjects and between unstimulated and stimulated saliva used. Mean staining for the subject group, at each cycle was always higher with unstimulated saliva compared to stimulated saliva and differences reached statistical significance at cycles 2-5. CONCLUSIONS: In vitro stain formation using unstimulated saliva from different individuals occurred at a faster rate and to a greater extent than when stimulated saliva from the same subjects was used, presumably reflecting differences in composition. Understanding the nature of these differences could provide fundamental information on the very poorly understood process of tooth staining. PMID- 12554121 TI - Single-step adhesives are permeable membranes. AB - OBJECTIVES: This study tested the hypotheses that micro-tensile bond strengths of all currently available single-step adhesives to dentine are adversely affected by delayed activation of a light-cured composite, and that such a phenomenon only occurs in the presence of water from the substrate side of the bonded interface. METHODS: In experiment I, a control three-step adhesive (All-Bond 2, Bisco) and six single-step adhesives (One-Up Bond F, Tokuyama; Etch&Prime 3.0, Degussa; Xeno CF Bond, Sankin; AQ Bond, Sun Medical; Reactmer Bond, Shofu and Prompt L-Pop, 3M ESPE) were bonded to sound, hydrated dentine. A microfilled composite was placed over the cured adhesive and was either light-activated immediately, or after leaving the composite in the dark for 20 min. In experiment II, three single-step adhesives (Etch&Prime 3.0, Xeno CF Bond and AQ Bond) were similarly bonded to completely dehydrated dentine using the same delayed light-activation protocol. In experiment III, a piece of processed composite was used as the bonding substrate for the same three single-step adhesives. The microfilled composite was applied to the cured adhesives using the same immediate and delayed light activation protocols. Bonded specimens were sectioned for micro-tensile bond strength evaluation. Fractographic analysis of the specimens was performed using SEM. Stained, undemineralised sections of unstressed, bonded specimens were also examined by TEM. RESULTS: When bonded to hydrated dentine, delayed light activation had no effect on the control three-step adhesive, but significantly lowered the bond strengths of all the single-step adhesives (p < 0.05). This adverse effect of delayed light-activation was not observed in the three single step adhesives that were bonded to either dehydrated dentine or processed composite. Morphological manifestations of delayed light-activation of composite in the hydrated dentine bonding substrate were exclusively located along the composite-adhesive interface, and were present as large voids, resin globules and honeycomb structures that formed partitions around a myriad of small blisters along the fractured interfaces. CONCLUSION: These features resembled the 'overwet phenomenon' that was previously reported along the dentine-adhesive interfaces of some acetone-based three-step adhesives. The cured adhesive layer in single-step adhesives may act as semi-permeable membranes that allow water diffusion from the bonded hydrated dentine to the intermixed zone between the adhesive and the uncured composite. Osmotic blistering of water droplets along the surface of the cured adhesive layer and emulsion polymerisation of immiscible resin components probably account for the compromised bond strength in single-step adhesives after delayed activation of light-cured composites. PMID- 12554122 TI - Letter to the editor: "The effect of acquired salivary pellicle on the surface free energy and wettability of different denture base materials" [C. Sipahi, N. Anil, E Bayramli Journal of Dentistry 29 (2001) 197-204]. PMID- 12554124 TI - Is there any rationale for prescribing hormone replacement therapy (HRT) to prevent or to treat osteoarthritis? AB - BACKGROUND: During the last two decades of the 20th century, hormone replacement therapy (HRT) has been considered as the sole pharmacological approach for counterbalancing or mitigating the effects of estrogens deprivation in post menopausal women. Subsequently, HRT has been widely recommended for the management of chronic diseases occurring, in women during the second half of their life. The overall risk/benefit ratio of estrogens has been recently reassessed in the light of long-term prospective studies failing to demonstrate the expected benefit of HRT on cardiovascular diseases incidence. Osteoarthritis (OA) is one of the chronic conditions for which HRT has been suggested to provide beneficial outcomes. RESULTS: The presence of estrogen receptors in human cartilage is no longer debated. However, cellular or animal models of OA do not provide an unequivocal pathway for the influence of gonadal steroids on cartilage. Similarly, studies attempting to correlate serum or urinary levels of sex steroids to the onset or progression of OA gave conflicting results. No randomized, prospective, controlled trial was designed to specifically assess the impact of hormone replacement therapy on symptomatic or structural progression of OA. Large-scale observational studies or trials designed to assess other potential benefits of estrogens suggest that HRT use does not provide symptomatic relief in OA but may interfere with its long-term structural progression, particularly in the lower limbs. CONCLUSION: Based on the recent results of the Women Health Initiative suggesting that HRT health risks may outweigh benefits, one can hardly recommend, with the current level of evidence, HRT as a first-line treatment against progression of OA. PMID- 12554125 TI - Molecular characterization and partial cDNA cloning of facilitative glucose transporters expressed in human articular chondrocytes; stimulation of 2 deoxyglucose uptake by IGF-I and elevated MMP-2 secretion by glucose deprivation. AB - OBJECTIVE: Recent evidence suggests that human chondrocytes express several facilitative glucose transporter (GLUT) isoforms and also that 2-deoxyglucose transport is accelerated by cytokine stimulation. The aim of the present investigation was to determine if human articular chondrocytes express any of the recently identified members of the GLUT/SLC2A gene family and to examine the effects of endocrine factors, such as insulin and IGF-I on the capacity of human chondrocytes for transporting 2-deoxyglucose. DESIGN/METHODS: PCR, cloning and immunohistochemistry were employed to study the expression of GLUT/SLC2A transporters in normal human articular cartilage. The uptake of 2-deoxyglucose was examined in monolayer cultured immortalized human chondrocytes following stimulation with TNF-alpha, insulin and IGF-I. Levels of MMP-2 were assessed by gelatin zymography following glucose deprivation of alginate cultures. RESULTS: Using PCR we detected transcripts for eight glucose transporter isoforms (GLUTs 1, 3, 6, 8, 9, 10, 11 and 12) and for a fructose transporter (GLUT5) in human articular cartilage. Expression of GLUT1, GLUT3 and GLUT9 proteins in normal human articular cartilage was confirmed by immunohistochemistry. The uptake of 2 deoxyglucose was dependent on time and temperature, inhibited by cytochalasin B and phloretin, and significantly accelerated in chondrocyte cultures stimulated with IGF-I. However, 2-deoxyglucose uptake was unaffected by short and long-term insulin treatment, which ruled out a functional role for insulin-sensitive GLUT4 mediated glucose transport. Furthermore, secretion of MMP-2 was increased in alginate cultures deprived of glucose. CONCLUSIONS: The data supports a critical role for glucose transport and metabolism in the synthesis and degradation of cartilage. PMID- 12554126 TI - New radiographic-based surrogate outcome measures for osteoarthritis of the knee. AB - OBJECTIVE: Minimum joint space width (mJSW) between the femoral condyle and tibial plateau is currently the principal radiographic outcome measure for knee osteoarthritis (OA). While mJSW measurement has been proven effective, there is a need for quantitative measures with greater sensitivity to OA so studies of disease modifying trials can be performed more quickly and with fewer subjects. Also, mJSW is a single outcome measure that does not reflect all disease changes. Here we present a study of new radiograph-based outcome measures to quantify OA progression. DESIGN: Our goal is made possible by a software algorithm that delineates both the edge of the tibial plateau and the femoral condyle on digitized knee radiographs. We also developed a new coordinate system based on anatomical landmarks to facilitate measurement of the outcome measures. We studied measurements of joint space width (JSW) and average JSW at fixed locations along the joint interface. The outcome measures were tested using duplicate acquisitions made from 16 OA and 18 normal knees. Reproducibility was quantified by the root-mean square standard deviation and the coefficient of variation. RESULTS: We found that the reproducibility for all outcome measures was better than or similar to that for mJSW. Despite a slightly higher reproducibility error for some new outcome measures compared to mJSW, they are potentially valuable since they reflect independent disease changes. CONCLUSION: We have demonstrated new reproducible radiographic outcome measures to quantify OA progression with the potential to be an improvement over standard minimum joint space width. PMID- 12554127 TI - Analysis of rabbit articular cartilage repair after chondrocyte implantation using optical coherence tomography. AB - OBJECTIVE: To evaluate the utility and limitations of optical coherence tomography (OCT) for immediate, high-resolution structural analysis of rabbit articular repair tissue following chondrocyte implantation without excising or sectioning the specimen. METHODS: Full thickness articular cartilage defects were created in the patellar grooves of 30 adult rabbit knee joints. Allogenic cultured chondrocytes embedded in collagen gels were implanted into the surgical defects. A periosteal patch was then sutured over the chondrocyte-collagen composites. Six animals per time point were sacrificed at 2, 4, 8, 12 and 24 weeks after surgery. The repair tissues were sequentially analysed by arthroscopic surface imaging, OCT, and histology. The resulting images were compared to determine qualitative and quantitative features of surface roughness, repair tissue integration, and micro-architecture. Statistical analysis was performed using Student's t -testing and linear regression. RESULTS: OCT was able to identify the bone and cartilage interface in normal rabbit articular cartilage and regenerated cartilage at 24 weeks post chondrocyte implantation. OCT was able to identify hypertrophy at 4 and 8 weeks, and subtle surface fibrillations at 24 weeks, comparable with histological analysis at low magnification (20x). More importantly, OCT was able to detect embedded gaps between the repair tissue and surrounding host cartilage. CONCLUSION: Close correlation was observed between OCT and histological analysis of morphological features important to the assessment of articular cartilage repair. These results demonstrate that OCT is capable of providing immediate 'optical biopsy' of the rabbit articular cartilage repair tissue without damaging the specimen, and suggest that this new technique, if integrated with an arthroscope, can potentially be used in longitudinal studies of articular cartilage repair in vivo. PMID- 12554128 TI - AG-041R stimulates cartilage matrix synthesis without promoting terminal differentiation in rat articular chondrocytes. AB - OBJECTIVE: AG-041R, a novel indolin-2-one derivative, has recently been demonstrated to induce systemic hyaline cartilage hyperplasia in rats. The aim of this study was to characterize its anabolic actions on chondrocytes. DESIGN: Chondrocytes were isolated from knee joints of 5-week-old SD rats. Effects of AG 041R on cartilage matrix synthesis were examined by measuring [(35)S]sulfate incorporation into proteoglycans, Alcian blue staining, and Northern blotting of cartilage matrix genes. ALP activity, mineral deposition and the expression of markers for hypertrophic chondrocytes, were assessed for terminal differentiation of chondrocytes. Roles of endogenous TGF-beta/BMPs and MEK1/Erk signaling in the action of AG-041R were investigated using the neutralizing soluble receptors and the MEK1 inhibitor. RESULTS: AG-041R accelerated proteoglycan synthesis assessed by both [(35)S]sulfate incorporation and Alcian blue stainable extracellular matrix accumulation. It also up-regulated the gene expression of type II collagen and aggrecan, as well as tenascin, a marker for articular cartilage. In contrast, AG-041R suppressed ALP activity, mineralization, and the gene expression of type X collagen and Cbfa1, indicating that AG-041R prevents chondrocyte terminal differentiation. AG-041R increased in BMP-2 mRNA, and the neutralizing soluble receptor for BMPs reversed the stimulatory effects of AG-041R on cartilage matrix synthesis. Moreover, AG-041R activated MEK1/Erk pathway, which was revealed to prevent chondrocyte terminal differentiation. CONCLUSION: AG-041R stimulates cartilage matrix synthesis without promoting terminal differentiation in rat articular chondrocytes, which is mediated at least in part by endogenous BMPs and Erk. The data demonstrates that AG-041R has a potential to be a useful therapeutic agent for articular cartilage disorders. PMID- 12554129 TI - Presence of pannus-like tissue on osteoarthritic cartilage and its histological character. AB - OBJECTIVE: To investigate and characterize pannus-like tissue which is often present on osteoarthritic articular cartilage. DESIGN: Cartilage specimens from 15 knee and five hip joints of patients with osteoarthritis (OA) undergoing arthroplasty were stained for HE and Safranin-O. They were also immunostained by antitype I collagen, type II collagen, CD68, IL-1beta and MMP3 antibodies. RESULTS: Ninety percent of joints have pannus-like tissue on the articular surface, preferentially in a marginal area. The articular cartilage was divided into three regions according to the location: the marginal zone, the intermediate zone and the paraeburnated zone. Pannuslike tissue in OA knee joint occurred 45.9%, 27.5% and 11.1% of the surface of each region respectively. Histologically, pannus-like tissue could be classified into the vascular type and the fibrous type. Extracellular matrix of both types of tissues were negative for Safranin-O and type II collagen, but positive for type I collagen. IL-1beta and MMP3 expressing cells are predominant in pannus-like tissue, whereas CD68 positive cells were infiltrated in only a few samples. Vascular type tissue kept continuity with bone marrow suggesting mesenchymal origin. CONCLUSION: Pannus like tissue exists in advanced OA cartilage, preferentially in the marginal zone. It expressed IL-1beta and MMP3, which strongly suggests that it contributes to cartilage degradation. PMID- 12554130 TI - Osteophyte development--molecular characterization of differentiation stages. AB - OBJECTIVE: Osteophytes are non-neoplastic osteo-cartilaginous protrusions growing at the margins of osteoarthritic joints. They can not only be considered as in situ repair tissue, but also represent an excellent in vivo model for induced cartilage repair processes. Our focus was to identify different steps of osteophyte development via analysis of expression patterns of marker genes of chondrocytic differentiation. DESIGN: We performed an extensive analysis of the presence and expression of matrix components using histochemical, immunohistochemical and in situ hybridization technology. RESULTS: Four different stages of osteophyte formation could be identified based on histomorphological and cell biological parameters: starting from mesenchymal condensates, chondrogenic differentiation is indicated by the onset of Col2A and aggrecan expression (stage I). Stage II shows fibrocartilage with an admixture of cartilaginous and fibrous matrix components such as Col2 and aggrecan on the one hand and Col1 on the other hand. The proliferating osteophyte (stage III) shows a zonal organization similar to the fetal growth plate cartilage with extensive chondrocyte hypertrophy in the zones next to ongoing endochondral bone formation. 'Mature' osteophytes (stage IV) resembled largely articular hyaline cartilage with a predominance of Col2 and aggrecan and Col6 found mainly pericellularily. CONCLUSIONS: The development of osteophytes is a good in vivo model to pursue chondrocyte differentiation from pluripotent mesenchymal cells to mature or hypertrophic chondrocytes in situ in the adult. The analysis of marker molecules of mesenchymal differentiation allows to identify different stages of repair tissue development and the transformation from fibrous tissue to neo-cartilage. Tissue architecture and matrix composition in mature osteophytes suggests that metaplastic neo-cartilagenous tissue might be one potential source of cartilage repair tissue in the adult joint. PMID- 12554131 TI - Kellgren & Lawrence grade 1 osteophytes in the knee--doubtful or definite? AB - The most universally accepted methods of classification of radiographic osteoarthritis (OA) is the Kellgren & Lawrence (K&L) grading system. However, there have been many criticisms of this including the grading of a doubtful osteophyte as a grade 1. We report the natural history of a given prevalent grade 1 osteophyte in a 10-year follow-up knee study. 62% of women graded at baseline with a doubtful osteophyte went on to develop true osteophytic knee OA compared with only 22% of controls with no sign of disease. So called 'doubtful' osteophytes appear to be 'real' and significantly related to OA knee and therefore cannot be ignored or classified as normal. PMID- 12554132 TI - Determination of the active metabolite of sibutramine by liquid chromatography electrospray ionization tandem mass spectrometry. AB - A sensitive and specific method for the determination of the active primary amine metabolite of sibutramine, N-di-desmethylsibutramine (BTS 54,505), in human plasma was developed, based on high-performance liquid chromatography (HPLC) electrospray ionization tandem mass spectrometry (MS-MS). The samples were extracted from plasma with methyl tert.-butyl ether, followed by separation and evaporation after addition of the internal standard, propranolol, and basification with sodium hydroxide. The residue was reconstituted in mobile phase and injected into the HPLC-MS-MS system. Chromatography was performed on an ODS MS column with a mobile phase consisting of acetonitrile (containing 0.1% trifluoroacetic acid, v/v)-0.1% trifluoroacetic acid (55:45, v/v) at a flow-rate of 0.3 ml/min. Multiple reaction monitoring using precursor-->product ion combinations at m/z 252.00-->125.00 and 260.00-->115.70 was applied to determine BTS 54,505 and propranolol, respectively. Linearity was confirmed in the concentration range 0.328-32.8 ng/ml in human plasma and the imprecision of this assay was less than 19.90% over the entire concentration range. The method is sufficiently sensitive and repeatable to be used in pharmacokinetic studies. PMID- 12554133 TI - Matrix-assisted laser-desorption time-of flight ionisation and high-performance liquid chromatography-electrospray ionisation mass spectral analyses of two glycosylated recombinant epoetins. AB - Mass spectrometric analyses of the recombinant proteins in Eprex and Aranesp were undertaken with the goal of producing reference mass spectra and evaluating strategies to improve its applicability as a method for equine and canine doping control of these substances. A simple, low chemical noise deglycosylation reaction removed microheterogeneity due to post-translational carbohydrate attachment and both proteins were detectable using MALDI-TOF-MS. Deglycosylated human erythropoietin (hEPO) was also detected using HPLC-ESI-MS. This is the first time that spectra of deglycosylated Eprex and Aranesp have been published. Eight synthetic reference standards, which match peptides produced by endoproteinase Glu-C enzymatic cleavage of Aranesp and/or Eprex, were analysed by ESI-MS and ESI-MS-MS. The E12 Glu-C peptide, common to both proteins, was detected at the low femtomole-level using gradient nano-HPLC-ESI-MS-MS in the positive ion mode. PMID- 12554134 TI - Newborn screening of homocystinuria: quantitative analysis of total homocyst(e)ine on dried blood spot by liquid chromatography with fluorimetric detection. AB - Identification of homocystinuric newborns is hindered by the pitfalls of neonatal screening programs. We propose a fluorimetric HPLC method with a rapid pre analytical step for homocysteine determination from neonatal dried blood spot cards. Homocysteine in blood spots sampled among 2000 healthy newborns on living day 4, averaged 2.92+/-2.07 microM (range 0.4-7.5). In eight homocystinuric control children, mean values were 61.71+/-52.84 microM (range 18.9-145.7). The method showed a good linearity (r=0.999), precision (RSD<7%) and recovery (95%). The correlation between blood spots and plasma samples was r=0.90. This method has all the essential features for a homocystinuria screening program: an easy and rapid pre-analytical step combined with method linearity and precision. PMID- 12554135 TI - Simultaneous determination of nimesulide and hydroxynimesulide in rat plasma, cerebrospinal fluid and brain by liquid chromatography using solid-phase extraction. AB - A liquid chromatographic method with UV detection for the quantification of nimesulide (N) and hydroxynimesulide (M1) in rat plasma, cerebrospinal fluid (CSF) and brain tissue is reported. Plasma samples (250 microl) and brain homogenates added with the right amount of the internal standard (I.S., 2' (cyclohexyloxy)-4'-nitrophenyl methanesulphonanilide, NS398) are extracted on C(18) disposable cartridges by solid-phase extraction (SPE), while CSF samples are analyzed without any extraction. The separation is performed at room temperature on a Waters Symmetry C(18) 3.5 microm (150x4.6 mm I.D.) column with acetonitrile-sodium citrate buffer pH 3.00 (53:47, v/v) as mobile phase, at a flow-rate of 1.1 ml/min and detection at 240 nm. The retention times are 3.3, 6.0 and 9.9 min for M1, N and I.S., respectively. The lower limits of quantitation for either nimesulide and M1 are 25 ng/ml for plasma, 20 ng/ml for CSF and 25 ng/g for brain tissue. The calibration curves are linear up to 10,000 ng/ml for plasma, 5000 ng/ml for CSF and 5000 ng/g for brain tissue. This new assay can be applied to the study of the role of nimesulide in the modulation of neuroinflammatory processes. PMID- 12554137 TI - Urinary concentrations of morphine after the administration of herbal teas containing Papaveris fructus in relation to doping analysis. AB - A quantitative method for the analysis of morphine in human urine in the concentration range between 0.25 and 2 microg/ml is described and validated. Morphine was determined after enzymatic hydrolysis of the urine. After liquid liquid extraction with dichloromethane-methanol (9:1) at pH 9.5, morphine was derivatized with N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) and analyzed with GC-MS (full scan). The limit of quantification of the method was 0.25 microg/ml. Two Papaveris fructus containing herbal teas were administered to five male volunteers and urine samples were taken quantitatively during the first 12 h after the administration. The morphine concentration in the tea was 10.4 and 31.5 microg/ml, respectively. Morphine was detected in the urine of all volunteers by 1 h after drinking the tea. Maximum morphine concentrations, 4.3 and 7.4 microg/ml, respectively, were obtained 4-6 h after administration. Doping positive urine samples were delivered for 1-9 h. PMID- 12554136 TI - Poly(ethyleneglycol) column for the determination of acetaminophen, phenylephrine and chlorpheniramine in pharmaceutical formulations. AB - New polar reversed-phase stationary phases in HPLC provide specific selectivities which can help to solve traditional chromatographic problems related to the development of chromatographic methods with widely different retention times for the sample components. One such case is the analysis of pharmaceutical formulations against the common cold. Acetaminophen, phenylephrine and chlorpheniramine, compounds with different polarities, are frequently associated in these drugs. An isocratic and rapid HPLC method for the simultaneous determination of the three compounds, acetaminophen, phenylephrine and chlorpheniramine, in capsules as pharmaceutical formulations, including the separation of impurities (4-aminophenol and 4-chloracetanilide) and excipients, has been developed and validated. The final chromatographic conditions employed a Supelco Discovery HS PEG column poly(ethyleneglycol) 15x0.46 cm, 5 microm. The mobile phase was 20 mM phosphate buffer, pH 7.0-acetonitrile (90:10, v/v) at a flow-rate of 1 ml/min. UV detection was performed at 215 nm for all the compounds except acetaminophen, which was measured at 310 nm. Validation parameters permit us to consider this method suitable. PMID- 12554138 TI - Measurement of paclitaxel in biological matrices: high-throughput liquid chromatographic-tandem mass spectrometric quantification of paclitaxel and metabolites in human and dog plasma. AB - A GLP-validated, sensitive and specific LC-MS-MS method for the quantification of paclitaxel and its 6-alpha- and 3'-p-hydroxy metabolites is presented. A 0.400 ml plasma aliquot is spiked with a (13)C(6)-labeled paclitaxel internal standard and extracted with 1 ml methyl-tert.-butyl ether. The ether is evaporated and the residue is reconstituted in 130 microl of 30% aqueous acetonitrile (ACN) containing 0.1% trifluoroacetic acid. Isocratic HPLC analysis is performed by injecting 50 microl of the reconstituted material onto a 50x2.1 mm C(18) column with an ACN-water-acetic acid (50:50:0.1) mobile phase at 200 microl/min flow. Detection is by positive ion electrospray followed by multiple reaction monitoring of the following transitions: paclitaxel (854>509 u), 6-alpha-hydroxy paclitaxel (870>525 u), 3'-p-hydroxy paclitaxel (870>509 u) and internal standard (860>509 u). Quantification is by peak area ratio against the 13C(6) internal standard. The method range is 0.117-117 nM (0.1-100 ng/ml) for paclitaxel and both metabolites using a 0.400 ml human or dog plasma sample. Analysis time per sample is less than 5 min. PMID- 12554139 TI - Optimization of protein precipitation based upon effectiveness of protein removal and ionization effect in liquid chromatography-tandem mass spectrometry. AB - Four categories of protein precipitation techniques (organic solvent, acid, salt and metal ion) were tested in plasma using spectrophotometry to assess protein removal efficiency across a range of volumes, species and lots. Acetonitrile, trichloroacetic acid (TCA) and zinc sulfate were found to be optimal at removing protein in their categories (>96, 92 and 91% protein precipitation efficiency at a 2:1 ratio of precipitant to plasma, respectively). A post-column infusion LC MS/MS system was used to assess ionization effect of a protein-bound drug caused by the endogenous components remaining after using various protein precipitants. The extent of ionization effect varied with mobile phase (-20 to 93%), protein precipitant (0.3-86%), but only slightly with species (86-93%). The optimal bioanalytical methodologies for removal of plasma proteins and minimal ionization effect for the probe molecule in positive ion turboionspray LC-MS/MS involve the use of TCA for precipitation with mobile phases consisting of either pure organic solvents (methanol:water or acetonitrile:water) or precipitation with all of the mass spectrometer compatible precipitants evaluated with a methanol:aqueous 0.1% formic acid mobile phase. PMID- 12554140 TI - Chiral separation and determination of R-(-)- and S-(+)-baclofen in human plasma by high-performance liquid chromatography. AB - The method presented here is a high-performance liquid chromatography (HPLC)-UV detection method for the determination of baclofen R-(-)- and S-(+)-enantiomers in human plasma using a chiral separation technique. Baclofen enantiomers were extracted from human plasma with a reversed-phase solid-phase extraction (SPE) cartridge. The extract was then injected onto a HPLC system with a UV detection system set at 220 nm. The separation was achieved by using a 150x4.6 mm, 5 microm Phenomenex chirex 3216 chiral column with a mobile phase consisting of 0.4 mM CuSO(4) in acetonitrile-20 mM sodium acetate (17:83). The calibration curves were linear for both R-(-)- and S-(+)-enantiomers of baclofen in the concentration range of 20-5000 ng/ml. The average regressions were 0.9980 and 0.9991 for R-(-)- and S-(+)-baclofen, respectively. Inter-day precision was 3.3-5.2% for R-(-) baclofen and 3.5-3.9% for S-(+)-baclofen at a concentration range of 60-4000 ng/ml. Intra-day precisions were 0.6-4.4 and 0.5-3.5% for R-(-)-baclofen and S (+)-baclofen, respectively. The average extraction recovery was 81.6% for R-(-) baclofen, 83.0% for S-(+)-baclofen and 94.0% for the internal standard (p aminobenzoic acid). The limit of quantitation for both R-(-)- and S-(+)-baclofen in human plasma was 20 ng/ml. The method is simple and easy to operate with accuracy and reproducibility and it is suitable for pharmacokinetic studies. PMID- 12554141 TI - Novel approach for the analysis of glycated hemoglobin using capillary isoelectric focusing with chemical mobilization. AB - In this work, a novel CIEF methodology for the analysis of the glycated hemoglobin, HbA(1c), in dimethylpolysiloxane coated fused-silica capillaries (DB 1, 50 microm I.D., 27 cm, 0.20 microm coating thickness), using a narrow pH ampholyte mixture (4% pH 6-8:pH 3-10, 10:1, v/v) in 0.30% methylcellulose, was developed. In the focusing procedure, a 0.100-mol l(-1) phosphoric acid solution was used as anolyte and a 0.040-mol l(-1) NaOH solution was used as catholyte. During method development, two types of mobilization of the focused hemoglobins were tested: pressure and chemical mobilization. Chemical mobilization performed better, allowing the complete baseline resolution of the hemoglobin of interest, HbA(1c), from its adjacent peak, HbA, in less than 8 min. In the chemical mobilization procedure, the catholyte was replaced by a 0.040-mol l(-1) NaOH solution containing 0.080 mol l(-1) NaCl. The proposed methodology was applied to the analysis of 31 hemolysate samples and validated with respect to the selectivity, inter-assay and intra-assay precision (both migration time and hemoglobin percentage concentration). In addition, HbA(1c) determinations were compared for the CIEF method and a chromatographic standardized procedure using cation-exchanger columns (Variant, Bio-Rad), adopted in a local clinical laboratory, showing excellent correlation (r(2)=0.872, n=31). The slope was found to be statistically equal to one but the intercept differed from zero. Also the Bland-Altman plot indicates bias, implying that the CIEF method yields HbA(1c) concentration higher than the reference method. The separation of the hemoglobins HbA, HbA(2), HbF and HbA(1c) and the variants HbS and HbC was also demonstrated (8 min run). The resolving power of the proposed CIEF method allowed baseline resolution of hemoglobins with a pI difference as small as ca. 0.03, as it is the case for the pairs HbC/HbA(2) and HbA/HbA(1c). PMID- 12554142 TI - Chromatographic system for the simultaneous measurement of plasma 18-hydroxy-11 deoxycorticosterone and 18-hydroxycorticosterone by radioimmunoassay: reference data for neonates and infants and its application in aldosterone-synthase deficiency. AB - A new chromatographic system for the steroid precursor separation and a sensitive radioimmunoassay system for the subsequent measurement of 18-hydroxy-11 deoxycorticosterone and 18-hydroxycorticosterone has been developed. 18-Hydroxy 11-deoxycorticosterone and 18-hydroxycorticosterone were extracted with methylene chloride and separated from cross-reacting steroids by Sephadex LH-20 column chromatography. Anti-18-hydroxy-11-deoxycorticosterone and anti-18 hydroxycorticosterone antibodies raised in rabbits were used. The lower detection limit of the assay is 0.03 nmol/l and 0.128 nmol/l for 18-hydroxy-11 deoxycorticosterone and 18-hydroxycorticosterone, respectively. Normal values for this assay in 128 healthy neonates and infants aged 0-5 months were established as a basis for the early hormonal diagnosis of aldosterone synthase deficiency types I and II. Its application for the diagnosis of aldosterone synthase deficiency is demonstrated in two patients with homozygous mutation/deletion in the encoding CYP11B2 gene. PMID- 12554143 TI - Determination of chromium in urine samples by complexation-supercritical fluid extraction and liquid or gas chromatography. AB - A method was developed for the determination of chromium as the Cr(acac)(3) complex in urine using SFE and chromatography. Quantitative extractions were achieved when the experiments were carried out under 3000 p.s.i. of pressure, at a temperature of 120 degrees C, with 2.0 ml of methanol, 30 min of static extraction and 5 min of dynamic extraction. The chromium was quantified by GC-FID detection. The calibration graph of Cr(acac)(3) solutions was linear between 0.50 and 43.0 microg ml(-1) of chromium (DL 0.18 microg ml(-1), R=0.9994). The same extracts were quantified by HPLC-340 nm detection. The calibration curve of the Cr(acac)(3) solutions was linear over a range of 0.013 to 60.0 microg ml(-1) (DL 0.02 microg ml(-1), R=0.9999). This method was applied to urine samples from 60 diabetic patients and 21 healthy volunteers. Chromium concentration ranges were 2.5-29.5 microg l(-1) for the diabetics and 5.9-12.3 microg l(-1) for the normal subjects. PMID- 12554144 TI - Determination of laquinimod in plasma by coupled-column liquid chromatography with ultraviolet absorbance detection. AB - Laquinimod is an immunomodulator that is currently in clinical trials. For pharmacokinetic and toxicokinetic studies in animals and humans a sensitive and accurate bioanalytical method was required. In this paper a bioanalytical method for the determination of laquinimod by liquid chromatography is described. After a protein precipitation step the plasma sample was injected onto a coupled-column HPLC system. After further purification from macromolecules on a short restricted access material C(18) column the analyte was transferred to a reversed-phase C(18) analytical column and separated from interfering substances. The analyte was detected by UV detection. The method was validated with respect to linearity, selectivity, precision, accuracy, limit of quantitation, limit of detection, recovery and stability. The limit of quantitation was 0.75 micromol/L, the intermediate precision was 1.8-3.6% (C.V.) and the accuracy was 97.7-114.7%. In conclusion, the method was found to perform well and is suitable for use in pharmacokinetic and toxicokinetic studies. PMID- 12554145 TI - Determination of corticosterone in mouse plasma by a sweeping technique using micellar electrokinetic chromatography. AB - The separation and on-line concentration of corticosterone in mouse blood was achieved by means of capillary electrophoresis/UV absorbance using sodium dodecyl sulfate (SDS) as a surfactant. The procedure involved the use of an on-line sample concentration method by sweeping-micellar electrokinetic chromatography (sweeping-MEKC). Optimal on-line concentration and separation conditions were determined. The detection limit for this method was 5 ng/ml (S/N=3) and photodiode array detection at 247 nm was used for identification. For the analysis of actual samples, corticosterones from blood samples of a non-stressed and stressed mouse were determined. The results show that only a minor amount of corticosterone was produced by a non-stressed mouse, whereas a significant amount was present in the blood sample from a stressed mouse. The method developed here can be used to examine corticosterone levels as a marker of stress in test animals and may also be used for estimating the effect of stress-release medications. PMID- 12554146 TI - Simultaneous determination of the monoamine neurotransmitters and glucose in rat brain by microdialysis sampling coupled with liquid chromatography-dual electrochemical detector. AB - A new type of chemically modified electrode based ring-disk electrode as the dual electrochemical detector (DECD) for high-performance liquid chromatography (HPLC) to simultaneously determine the monoamine neurotransmitters and glucose is described. The ring electrode was modified with an ion-exchange polymer overoxidized polypyrrole (OPPy) and the disk electrode was modified with nano Au colloid and glucose oxidase (GOD). The electrochemical behaviors of dopamine (DA) and ascorbic acid (AA) at the OPPy chemically modified electrode (CME) were investigated by differential pulse voltammetry (DPV). It was found that the CME could permeate dopamine cations and repelled the ascorbate anions, which could be used to determine the monoamine neurotransmitters and avoid the interference of AA. The electrochemical behavior of glucose at the Nafion/GOD-Au colloid/GC CME was investigated by amperometry and flow injection analysis (FIA). It was found that the sensitivity of the CME increased apparently in determination of glucose. In order to obtain better separation and current responses of the analytes in HPLC-DECD, several operational parameters have been investigated. Under the optimum conditions, the method showed good stability and reproducibility. The application of this method coupled with microdialysis sampling for in vivo simultaneous determination of monoamine neurotransmitters and glucose in rat brain was satisfactory. PMID- 12554147 TI - Improved method for plasma malondialdehyde measurement by high-performance liquid chromatography using methyl malondialdehyde as an internal standard. AB - Measurement of malondialdehyde (MDA) is an important contribution to the assessment of oxidative stress. We report a sensitive and reproducible high performance liquid chromatography (HPLC) method for measurement of plasma MDA in the assessment of lipid peroxidation. Using methyl malondialdehyde (Me-MDA) as an internal standard with reversed-phase HPLC and UV detection and derivatisation with 2,4 dinitrophenylhydrazine (DNPH), we obtained maximum MDA values with 60 min incubation of 10% plasma with 1 M NaOH at 60 degrees C. The dilution of the plasma and a longer incubation time in the alkaline hydrolysis step greatly improved recovery of MDA from its bound form. Ratios of peak height of MDA/Me-MDA were linear over a range of 0-100 microM with correlation coefficients >0.99. The recovery was 88.5%. Within and between run variations were <4 and <7%, respectively. The mean MDA value measured in 20 healthy volunteers was 13.8 microM (+/-1.32). PMID- 12554148 TI - Liquid chromatography for iothalamate in biological samples. AB - We have previously reported an iothalamate assay for the assessment of the glomerular filtration rate (GFR) that required a long column equilibration time and 22 min run time per sample. We now report a simpler assay that requires a run time of only 5.5 min and is more precise and accurate than the earlier technique. The mobile phase consisted of methanol-acetonitrile-50 mM sodium monobasic phosphate (10:5:85, v/v) at pH 4.4, pumped at a rate of 1.5 ml/min on a C(18) reversed-phase column. Samples of plasma and urine were deproteinized with 1 volume of 4% perchloric acid or 9 volumes of 2% perchloric acid, respectively. No internal standard was used. The diode array detection system collected absorbance at 240 nm and the peak height areas of iothalamate were determined. The iothalamate peak appeared at 3.5 min. Detector response was linear over the range tested (10-2000 microg/ml). Within-run precision was <3% for both plasma and urine and accuracy was 96-102%. Between-day precision for plasma and urine analyses were <7%. The recovery of iothalamate in urine and plasma were 102% and 91%, respectively. There was excellent thermal and pH stability of iothalamate. No interference was found with para-amino hippuric acid (PAH) or N-acetyl PAH, which can be simultaneously assayed, if desired. PMID- 12554149 TI - Determination of serum thyroxine enantiomers in patients by liquid chromatography with a chiral mobile phase. AB - A chromatographic method for the separation and determination of D- and L thyroxine enantiomers (D-, and L-T4) in human serum with a chiral ligand ion exchange system using a chiral mobile phase additive and a silica column was established. An aqueous eluent containing L-proline (L-pro) sufficiently complexed copper II ions and triethylamine (TEA) was used. It was monitored with a UV detector. The separation was completed in 12 min. The method has acceptable sensitivity, precision and accuracy for analysis. The limit of detection and the limit of quantitation for both D- and L-T4 were 0.1 microg/ml and 0.8 microg/ml, respectively. Calibration curves were linear within 1-100 microg/ml; the mean correlation coefficients were r(D-T4)=0.9986 for D-T4 and r(L-T4)=0.9978 for L T4. T4 enantiomers were separated on baseline under the optimum condition. L-T4 eluted before D-T4. The concentration of D-T4 and L-T4 in 45 thyroid patients serum (hyperthyroid, hypothyroid, thyroidectomy, goitre or thyroiditis) using HPLC was determined, those results showed that D,L-T4 concentration varied in different thyroid patient. Attention should be paid to this result in treating thyroid disease in the clinic. PMID- 12554150 TI - Membrane chromatographic method for the rapid purification of vitellogenin from fish plasma. AB - A membrane chromatographic method was developed for the rapid purification of vitellogenin (Vtg) from the plasma of 17beta-estradiol induced loach (Misgurnus angaillicaud atus) and carp (Cyprinus carpio). The time required for the proposed procedure is less then 10 min at a flow-rate of 5 ml/min of the mobile phase, and 0.5 ml of fish plasma could be separated in one cycle. Multistep gradient elution was more suitable for the separation than linear gradient elution. Under optimized conditions, a single Vtg peak can be obtained and its identity was confirmed by SDS-PAGE and gel-permeation chromatography assessment. This method is rapid and easy to operate compared to conventional HPLC and FPLC columns for Vtg separation. PMID- 12554151 TI - Determination of Simvastatin in human plasma by liquid chromatography-mass spectrometry. AB - A simple, sensitive and selective liquid chromatography coupled with electrospray ionization mass spectrometry (LC/ESI/MS) method for the determination of simvastatin (I) has been developed. After extraction by ethyl acetate, using lovastatin (II) as internal standard, solutes are separated on a C(18) column with a mobile phase consisting of methanol-water (9:1). Detection is performed on an atmospheric pressure ionization single quadruple mass spectrometer equipped with an ESI interface and operates in positive ionization mode. Simvastatin quantification was realized by computing peak area ratio (I/II) of the extracts analyzed in SIM mode (m/z: 441 and m/z: 427 for I and II, respectively) and comparing them with calibration curve (r=0.9997). Accuracy and precision for the assay were determined by calculating the intra-batch and inter-batch variation at three concentrations 0.1, 5.0, 10.0 ng/ml; the intra batch relative standard deviation (RSD) was less than 10% and ranged from 1.8 to 8.5%, respectively; the inter-batch RSD was less than 20% and ranged from 4.1 to 16.5%. The limit of detection was 0.05 ng/ml. PMID- 12554152 TI - Determination of tenofovir in human plasma by high-performance liquid chromatography with spectrofluorimetric detection. AB - A sensitive high-performance liquid chromatography method was developed with spectrofluorimetric detection for the determination of tenofovir, a new HIV reverse transcriptase inhibitor, in human plasma. After precipitation of 200 microl of plasma samples by methanol and evaporation of the supernatant, fluorescent derivatized compounds were obtained by a 40-min incubation at 80 degrees C with chloroacetaldehyde 0.34% at pH 4.5. The assay was performed isocratically using 5 mM Na(2)HPO(4) (pH 6), containing tetrabutylammonium (TBA) chloride 5 mM, and acetonitrile (85:15, v/v) as mobile phase, and a Cluzeau C(8) plus satisfaction column maintained at 35 degrees C. Detection was performed at excitation and emission wavelengths set at 236 and 420 nm, respectively. In these conditions, tenofovir can be separated from adefovir, the internal standard, and endogenous substances. The method was found to be linear and has been validated over a concentration range of 5-1000 microg/l. The average coefficient of the limit of quantification (5 microg/l) was 5.38% and at this concentration, a signal-to-noise ratio of 500 was measured PMID- 12554153 TI - A simple and sensitive HPLC method for determination of gliclazide in human serum. AB - A simple, rapid and specific method for analysis of gliclazide in serum by a sensitive high-performance liquid chromatographic method is described. Only 100 microl of serum and a little sample work-up is required. A simple procedure of extraction by toluene followed by evaporation to dryness under a gentle stream of air and dissolving the dried residue in mobile was used. The gliclazide peak was separated from endogenous peaks on a C(8) column by a mobile phase of acetonitrile-water (45:55, v/v), pH 3. Gliclazide and internal standard (phenytoin) were eluted at 6.8 and 3.8 min, respectively. The limit of quantitation (LOQ) for gliclazide in serum was 75 ng/ml at 230 nm. The method was linear over the range of 75-10,000 ng/ml with r(2) of 0.999. Mean recovery for gliclazide and internal standard was 84.5 and 87%, respectively. PMID- 12554240 TI - Surgical induction of ovulation in polycystic ovarian disease. AB - The current lines of treatment of an ovulatory infertility due to polycystic ovarian disease lie between weight loss, insulin sensitising agents, clomiphene citrate, gonadotrophin therapy, or finally ovarian cauterisation. This review is looking at some of the current evidence for surgical and medical lines of treatment. PMID- 12554241 TI - Scottish female obstetricians' views on elective caesarean section and personal choice for delivery. AB - We have been reminded many times in the medical and lay press about the views of London female obstetricians choosing elective caesarean section for non-medical reasons. This paper asked Scottish female obstetricians for their personal choice regarding delivery and showed that only 15.5% would choose elective caesarean section compared with 31% and 21% in two surveys of London female obstetricians. None of the women in this group who had had a vaginal delivery would choose elective section. PMID- 12554242 TI - Endothelin-1 activity in pregnancy. AB - Plasma endothelin-1 activity was measured by radioimmunoassay in 24 normotensive non-pregnant women and in 24 normotensive pregnant, 24-aproteinuric hypertensive and 24 pre-eclamptic women. Endothelin-1 activity was increased in the pre eclamptic group (2.7 +/- 06 pg/ml) compared to the normotensive non-pregnant (1.0 +/- 0.8 pg/ml; P < 0.0001), normotensive pregnant (1.2 +/- 0.4 pg/ml; P < 0.0001) group and the aproteinuric hypertensive group (1.4 +/- 0.7 pg/ml; P < 0.0001). There was no difference in endothelin-1 activity between the normotensive non pregnant and normotensive pregnant group (1.0 +/- 0.8 vs. 1.2 +/- 0.4 pg/ml; P = 0.3). However, there was a difference between the aproteinuric hypertensive group (1.4 +/- 0.7 pg/ml) and both the normotensive nonpregnant (1.0 +/- 0.8 pg/ml; P < 0.01) and the normotensive pregnant group (1.2 +/- 0.4 pg/ml; P < 0.06). The birth weight in the pre-eclamptic group (2.48 +/- 0.61 kg) was significantly lower than that of the normotensive pregnant group (2.85 +/- 0.33 kg; P < 0.001) and the aproteinuric hypertensive group (2.99 +/- 0.46 kg; P < 0.001). In addition, there was no difference in birth weight between the normotensive pregnant group and aproteinuric hypertensive group (2.85 +/- 0.33 vs. 2.99 +/- 0.46 kg; P = 0.3). A significant Pearson's correlation of plasma endothelin-l versus birth weight in the pre-eclamptic group was obtained (r = 0.64; P < 0.0001). Endothelin-1 activity is increased with pre-eclampsia in black African women with pre-eclampsia. The results of our study also suggests an ethnic difference in plasma endothelin-1 activity when compared to result of studies carried out in Caucasian women with hypertensive disorders of pregnancy. PMID- 12554243 TI - The relative risk of 'fetal distress' in pregnancy associated with meconium stained liquor at different gestation. AB - Pregnancies complicated with meconium-stained liquor are regarded as at risk of fetal distress. Evidence from animal and human studies indicate that passage of meconium may be related to fetal maturity. This retrospective study attempts to quantify the relative risk of non-reassuring cardiotocography or 'fetal distress' in pregnant women complicated by meconium-stained liquor for preterm, term and post-term pregnancies. A total of 9542 singleton pregnancies, delivered in a tertiary obstetric unit in Hong Kong between 1 July 1996 and 31 June 1999, were included in the study. Of these pregnancies, 1946 (20.4%) were identified as having meconium-stained liquor, ranging from thin to thick staining. There was a strong association between incidence of meconium-stained liquor (P < 0.0005) and moderate/thick meconium-stained liquor with advanced gestational age. The incidence of non-reassuring cardiotocography in women presenting with meconium stained liquor was significantly higher (9.8% vs. 6.4%). The relative risk of non reassuring cardiotocography in women with meconium-stained liquor increased with more advanced gestation. Close fetal surveillance during labour is required among these pregnancies. Premature labour is associated with higher incidence of fetal distress but the presence of meconium did not pose an additional risk. PMID- 12554244 TI - Barriers to the use of antenatal and obstetric care services in rural Kano, Nigeria. AB - The objective of this study was to identify the sociocultural and economic factors that act as barriers to women's use of antenatal care services and hospital delivery in a rural community in Kano State, Northern Nigeria. The study was based on an interview of 107 pregnant women conducted by a trained midwife in the native language of the area. Findings indicate that the majority of women (88%) (CI = 81.8-94.2%) in the study area did not attend for antenatal care, and 96.3% (CI = 93.0-99.8%) had delivered or plan to deliver at home without a skilled attendant. Major barriers identified were economic, cultural and those related to the women's perception of their condition. The study recommends that poverty reduction and economic empowerment of rural women are prerequisites for any tangible improvement in the utilisation of antenatal care and obstetric delivery services. PMID- 12554245 TI - Usefulness of risk scoring at booking for antenatal care in predicting adverse pregnancy outcome in a rural African setting. AB - Antenatal care (ANC) attempts to screen and provide surveillance and treatment to individuals according to the level of need. We assessed the value of antenatal risk allocation made at the first visit in identifying women who will experience pregnancy complications in a rural area in Zimbabwe. As part of an ANC trial women were allocated into low- and high-risk categories based on medical, demographic and obstetric history. All highrisk women were recommended hospital delivery. This evaluation is based on 5223 women who received traditional care from nurse-midwives in 12 rural health centres, of whom 2890 (55%) were classified as high risk by the traditional risk markers, including 1618 nulliparous women. Complications occurred in 924 (17.7%) women 577 (62.4%) of whom had risk markers identified at booking. Twenty per cent (577/2890) of women classified as high risk developed complications. There was a high recurrence of complications, such as hypertensive disorders, operative delivery and preterm delivery. Nulliparity was a risk for low birth weight, operative delivery and hypertensive disorders, whereas grandmultiparity (> or =6) was a risk for hypertension in pregnancy. Young age (< or =16 years) was)a risk factor for low birth weight and perinatal death. Age above 35 years was not an independent risk factor. The traditional risk allocation system, with a likelihood ratio of 1.16, was not effective in identifying women at risk of pregnancy complications and resulted in too large a risk group for referral. PMID- 12554246 TI - Prevalence and risk factors for anaemia in pregnancy in South Southern Nigeria. AB - The aim of this study was to ascertain the prevalence of anaemia in pregnancy in South Southern Nigeria, and to identify an 'at-risk' group for targeted intervention. Antenatal women were screened for anaemia for 1 year (October 2000 September 2001) at the booking clinics of our three government hospitals in Kwale zone. Univariate and multivariate logistic regressions of risk factors for anaemia ([Hb] < 11 g/dL) and severe anaemia ([Hb] < 7.0 g/dL) were carried out. The prevalence of overall anaemia and severe anaemia was found to be 56.1% and 6.7%, respectively. Independent risk factors for anaemia and severe anaemia were primigravidity, booking in late pregnancy and wet season. We conclude that anaemia remains a common problem in our population, and additional intervention for the 'at-risk' group is recommended. PMID- 12554247 TI - No benefit, but increased harm from high dose (100 microg) misoprostol for induction of labour: a randomised trial of high vs. low (50 microg) dose misoprostol. AB - Misoprostrol, a synthetic analogue of prostaglandin E(1), has been used for cervical preparation. Its ideal dose, route and frequency of administration are still under investigation. We conducted a randomised controlled trial, in a tertiary hospital in a developing country, to compare misoprostol 50 microg (low) and 100 microg (high) for effectiveness and safety in induction of labour at term. Women admitted for induction of labour with a singleton live fetus in cephalic presentation after 37 weeks' gestation were recruited. A misoprostol tablet was inserted in the posterior vaginal fornix at 8-hour intervals. Main outcomes were duration of induction, maternal and fetal complications. The mean duration of induction was 15.4 (SD 10.6) and 14.2 (SD 13.6) h in the low- and high-groups respectively (P = 0.095). There was no difference in need for augmentation with oxytocin (OR 0.82; 95% Cl 0.36-1.86) or operative delivery (OR 1.29; 95% CI 0.26-6.84). There were two uterine ruptures and four intrapartum stillbirths in the high misoprostol group. There was no difference in postpartum haemorrhage, 9.5% vs. 7.9% (P = 1.00) and admissions to the neonatal unit 18.8% vs. 17.0% (P = 0.980) in the 1ow- and high-groups) respectively. Misoprostol 50 microg was as effective as the 100 microg dose for induction of labour whereas the higher dose had an increased risk of serious complications. PMID- 12554248 TI - Predictors of obstetric intervention rates: case-mix, staffing levels and organisational factors of hospital of birth. AB - We performed a cross-sectional study of all Thames maternity units, 1994-96, including 540,834 live and stillbirths. In contrast to recent media speculation, no association of caesarean section rates with midwifery staffing levels was found after adjustment for confounders. The only association with staffing was with levels of junior obstetric staffing, which could be a reflection of less experienced management of labour. Caesarean section rates were also associated positively with the levels of delivery beds, which could be a reflection of the closer monitoring of labour that may result from increased bed availability. Both caesarean section and instrumental vaginal delivery rates were associated with epidural rates, which was expected from the literature. Variations in epidural rates were mainly associated with variations in demographic case-mix, due possibly to patient demand. Demographic case-mix was also associated with instrumental vaginal deliveries but not the caesarean section rate. PMID- 12554249 TI - Exploring the causes of and risk factors for maternal deaths in a rural Nigerian referral hospital. AB - A 10-year (1990-9) review of all maternal deaths at Mater Misericordiae Hospital is presented. A total of 104 deaths occurred out of 13,391 deliveries (MMR 777/100,000). Haemorrhage, sepsis and Hypertensive diseases in pregnancy were the leading causes of death. Age below 15, Nulliparity, Grandmultiparity, low socio economic class, lack of formal education and being unbooked were also significant risk factors. Delay in accessing health facility was also an identified contributing factor. Public enlightenment to encourage utilisation of antenatal facilities, prompt referral of complicated cases and adequate equipping of the referral hospital is suggested. PMID- 12554250 TI - Retained placenta continues to be fatal but frequency can be reduced. AB - Retained placenta is potentially lift-threatening not only because of retention per se, but because of associated haemorrhage and infection as well as complications related to its removal. These risks are increased in women in poor social circumstances due to pre-existing malnutrition, anaemia and unsupervised home deliveries. The present study was undertaken to assess the situation in this rural area. A previous study at the same place suggested preventive strategies an and individualised approach. Incidence of retained placenta has been 0.23% of all births over 15 years. Of the deliveries at Kasturba Hospital, the incidence of retained placenta was 0.008% (two of 23,838 vaginal deliveries). Sixty-five women were admitted with retained placenta after home deliveries and three after delivery at other hospitals The age of most of the patients was between 20 and 29 years and most were para 2 or 3. In twenty-three (32.4%) cases, women had delivered preterm. Twelve (16.9%) women had previous uterine surgery and 10 (14.1%) had had a retained placenta in the past. Twenty-six (36.61%) women had come in a state of severe shock. Thirty-six (50.7%) women required general anaesthesia for manual removal. One woman with an adherent placenta had to undergo hysterectomy (1.40%). The maternal mortality was 5.6%. Perinatal loss was 7.04%. It is unfortunate that of the four deaths, two women had actually delivered at a near by district hospital and were referred moribund and died. A properly conducted delivery can reduce the incidence of retained placenta and if retention occurs, timely appropriate treatment can save life. PMID- 12554251 TI - Outpatient cervical ripening prior to induction of labour. PMID- 12554252 TI - Early onset, severe pre-eclampsia and HELLP syndrome: sex ratio of infants. PMID- 12554253 TI - Prepregnancy counselling for women with medical disorders. PMID- 12554254 TI - Correlates of knowledge of antenatal patients about HIV transmission in Ibadan, Nigeria. PMID- 12554255 TI - Infertility in Shanghai: prevalence, treatment seeking and impact. AB - To assess the level of treatment seeking and impact on marriage of infertility among newly married couples of Shanghai, China, a total of 7872 newly married couples, enrolled between 1987 and 1988, were followed-up for 5 years. The prevalence of infertility (no fertile conception after 24 months of unprotected intercourse) was found to be 5.1%. Couples' age at marriage, education, prior induced abortion, miscarriage, use of IUD or hormonal contraceptives and medical problems of both partners were associated significantly with infertility. About 57% of infertile couples sought infertility treatment and subsequently had a higher probability of childbirth (42%) than non-seekers (28%). Husband's education, induced abortion and both partner's previous medical problems were associated significantly with infertility treatment seeking. Moreover, infertile couples were 2.2 times more likely to divorce than their counterparts (95% CI 1.52-3.18). We conclude that infertility in Shanghai is modest, but a substantial number of infertile couples would not like to seek infertility treatment. Further research is needed on this subject. PMID- 12554256 TI - Serum levels of inhibin B in men and their relationship with gonadal hormones, testicular volume, testicular biopsy results and sperm parameters. AB - Currently, serum inhibin B levels, indicating testicular function due directly to its testicular origin, has been used increasingly in assisted reproductive units. Our aim in this prospective study was to evaluate the biological significance of inhibin B in gonadal dysfunctional males and the usefulness of inhibin B for the detection of male reproductive dysfunction. We included 52 oligoazoospermic and 20 normospermic men in this study. In our study serum inhibin B levels had statistically significant negative correlation with serum FSH and LH levels (P < 0.001, r: 0.781) and statistically moderate positive correlation with oestradiol levels (P < 0.005, r: 0.292). Inhibin B levels had significantly positive correlation with sperm count (P < 1.005, r: 0.851) and with testicular volume (P < 0.001, r: 0.466). Consequently, serum inhibin B level determination is a useful and non-invasive method for the evaluation of male gonadal dysfunction, taking into account its correlation with history, clinical examination, hormonal parameters, testicular volume, spermiogram and testicular biopsy. PMID- 12554257 TI - Sexual dysfunction among infertile Nigerian women. AB - The sexual histories of 97 infertile women were collected using questionnaires. Sexual dysfunction was found to be common among them, particularly frigidity (78.4%), dyspaereunia (57.7%), difficulty with sexual arousal (20.6%) and difficulty in achieving orgasm (20.6%). These could affect sexual acceptance and subsequently coital frequency, thus complicating their infertility problem further. Hence, there is the need of addressing these issues when managing these patients. PMID- 12554258 TI - Vaginal sacrospinous fixation: experience in a district general hospital. AB - This is an observational retrospective study, which included 77 patients who underwent sacrospinous fixation (SSF) over a period of 3 years in a district general hospital. Casenotes were reviewed and all patients were invited to attend a further review appointment (14-49 months). The object was to determine short- and long-term success of SSF,particularly in elderly (28% were > or = 70 years), obese (24.7%) and medically compromised (64.9%) patients. Fifty-two patients (67.5%) had previous gynaecological procedures, while the remaining 25 (32.5%) had no previous surgery. Intra- and postoperative complications were 3.9% and 32.5%, respectively. There was considerable relief of patients' symptoms at both short- and long-term followup, while the rate of recurrence of vault prolapse was 10.3%. Our experience suggests that SSF appears to be a safe and effective procedure, especially for those who may constitute surgical or anaesthetic risks. It has a reasonable success rate, good postoperative recovery and acceptable long term results. PMID- 12554259 TI - Feasibility of performing TVT operation for stress urinary incontinence under general anaesthesia. AB - This study assesses the feasibility of performing the tension-free vaginal tape (TVT) operation under general anaesthesia (GA). Fifty patients underwent TVT operation under GA. The mean age was 51.3 years (range 32-80). The mean operating time was 21 minutes (range 15-24). Forty (80%) were primary and 10 (20%) were secondary procedures. Postoperatively 22 (44%) patients passed urine within 24 hours and an indwelling catheter was used in 28 (56%) patients. The mean stay was 40 hours (8 hours-5 days). Postoperative evaluation was performed at 2, 6 and 11 months (median follow up 6 months). One patient had an infected retropubic haematoma, necessitating removal of the sling. Of the 49 patients analysed, 72% were completely cured and another 20% were significantly improved. Our findings show that TVT operation can be performed under GA without intra-operative continence demonstration. The success rate of the operation is comparable with the published results. PMID- 12554260 TI - The effect of platinum-based combination chemotherapy on the lymph nodes in advanced-stage epithelial ovarian cancer: does it decrease the incidence of lymph node involvement? AB - This paper aims to determine the impact of platinum-based combination chemotherapy on the lymph nodes in advanced-stage epithelial ovarian carcinoma. From 1997 to 2000, the patients in whom we performed lymphadenectomy before (group A) or after chemotherapy (group B) in the Department of Obstetrics and Gynecology, Akdeniz University School of Medicine were enrolled in this study. A total of 47 cases were included in the study. Twenty five cases had lymphadenectomy during the initial laparatomy and 22 cases during second-look procedures. Lymph node metastasis was detected in 14 (56%) patients in group A and in 10 (45.4%) cases in group B (P > 0.05). Platinum-based combination chemotherapy does not decrease significantly the incidence of involved lymph nodes in advanced-stage epithelial ovarian carcinoma. PMID- 12554261 TI - Medical termination of pregnancy at 9-12 weeks of gestation. AB - Current RCOG guidelines advise that surgical termination should be offered to those within the 9-12 weeks gestation band. While auditing the quality of services offered for termination of pregnancy in our unit, it became apparent that many women presenting at this gestation were requesting a medical method. There has been little clinical research into medical method of abortion at this gestation. The aim of the study was to assess the efficacy of medical methods of termination at 9-12 weeks gestation. A retrospective analysis of 25 cases who underwent medical termination using a regime of mifepristone followed 48 hours later by a course of vaginal gemeprost was undertaken. Complete abortion was achieved in 96% of cases; 92% of women required no more than two pessaries to achieve complete abortion. All but one patient was suitable for discharge on the same day. One woman underwent surgical evacuation in view of heavy bleeding. We conclude that medical TOP is a safe alternative to surgical method at 9-12 weeks' gestation. PMID- 12554262 TI - Current practice of hysterectomy and oophorectomy in the United Kingdom and Republic of Ireland. AB - The purpose of this study was to estimate current surgical practice of hysterectomy and prophylactic oophorectomy of UK and Irish consultant gynaecologists. Individual and regional variation in surgical practice and factors influencing surgical practice were assessed. A postal questionnaire was sent to all 1536 consultants in obstetrics and gynaecology currently practising in the United Kingdom and Ireland, with a 52.7% response rate. Approximately 60% of hysterectomies were abdominal, 37% vaginal and 4% laparoscopic-assisted, with junior consultants performing a greater proportion of hysterectomies vaginally. There was a wide variation in method of hysterectomy, both individually and regionally. Only 21% of consultants would routinely consider performing prophylactic oophorectomy and of these the majority would perform this procedure in women aged between 46 and 50 years old. This large variation in surgical practice indicates a need to form a consensus on optimal surgical techniques and to ensure adequate surgical training for all gynaecologists. PMID- 12554263 TI - Macroamylasaemia in pregnancy. PMID- 12554264 TI - Late puerperal sagittal sinus thrombosis associated with primary antiphospholipid antibody syndrome. PMID- 12554265 TI - Zavanelli manoeuvre in a patient with a previous caesarean scar. PMID- 12554266 TI - Urinary bladder leiomyoma during pregnancy: report of one case with immunohistochemical studies. PMID- 12554267 TI - Simultaneous intrapartum uterine and bladder rupture. PMID- 12554268 TI - A case of Ogilvie's syndrome after caesarean section. PMID- 12554269 TI - Silent uterine rupture following second trimester medical termination of pregnancy in a woman with an artificial urinary sphincter and three previous caesarean sections. PMID- 12554270 TI - Undiagnosed placenta percreta. PMID- 12554271 TI - Placenta percreta revisited. PMID- 12554272 TI - Haemoperitoneum due to placenta percreta in the third trimester mimicking placental abruption. PMID- 12554273 TI - 'Classical' caesarean section at or near term in the morbidly obese obstetric patient. PMID- 12554274 TI - Abnormal Down's screening test associated with severe hyperemesis gravidarum. PMID- 12554275 TI - Rare cause of irregular bleeding in a 12-year-old girl. PMID- 12554276 TI - Transverse leukonychia secondary to paclitaxel-carboplatin chemotherapy in a patient with ovarian cancer. PMID- 12554277 TI - Endometrial polyp and the levonorgestrel intrauterine system--a case report and literature review. PMID- 12554278 TI - Cloacal defect in a 23-year-old with 47,XXX karyotype and clinical features of Cat Eye syndrome. PMID- 12554279 TI - Suprarenal vena caval thrombosis and pulmonary embolism associated with a benign ovarian cyst. PMID- 12554280 TI - Unusual Trousseau's syndrome in ovarian carcinosarcoma: multiple systemic thromboembolic events. PMID- 12554281 TI - Urethral diverticulum presenting as anterior vaginal wall cyst. PMID- 12554282 TI - Cervical stump carcinoma following subtotal hysterectomy. PMID- 12554283 TI - Abnormal glandular smear: first evidence of ovarian malignancy. PMID- 12554284 TI - Inadvertent bladder injury due to tumour infiltration during radical hysterectomy procedure for cervical cancer: complete healing with postoperative chemotherapy. PMID- 12554285 TI - Re: Bell-Thomas and Weiner evaluation of ventouse failures in a district general hospital. PMID- 12554286 TI - Surgical correction of genuine stress incontinence in an achondroplastic woman. PMID- 12554291 TI - Editorial. PMID- 12554292 TI - The benefits of 3D modelling and animation in medical teaching. AB - Three-dimensional models created using materials such as wax, bronze and ivory, have been used in the teaching of medicine for many centuries. Today, computer technology allows medical illustrators to create virtual three-dimensional medical models. This paper considers the benefits of using still and animated output from computer-generated models in the teaching of medicine, and examines how three-dimensional models are made. PMID- 12554293 TI - Image quality, compression and segmentation in medicine. AB - This review considers image quality in the context of the evolving technology of image compression, and the effects image compression has on perceived quality. The concepts of lossless, perceptually lossless, and diagnostically lossless but lossy compression are described, as well as the possibility of segmented images, combining lossy compression with perceptually lossless regions of interest. The different requirements for diagnostic and training images are also discussed. The lack of established methods for image quality evaluation is highlighted and available methods discussed in the light of the information that may be inferred from them. Confounding variables are also identified. Areas requiring further research are illustrated, including differences in perceptual quality requirements for different image modalities, image regions, diagnostic subtleties, and tasks. It is argued that existing tools for measuring image quality need to be refined and new methods developed. The ultimate aim should be the development of standards for image quality evaluation which take into consideration both the task requirements of the images and the acceptability of the images to the users. PMID- 12554294 TI - The facial reconstruction of an Ancient Egyptian Queen. AB - The National Museums of Scotland Mummy Project has provided important new information about a burial excavated in Egypt. This has resulted in the facial reconstruction of a woman who was probably a queen at Thebes ca. 1570-1520 BCE. There are strong suggestions from the grave goods and her diet that this woman may have been ethnically Nubian rather than Egyptian. However, it is not yet possible to establish her ethnic identity for sure, so a definitive reconstruction of her appearance in life remains elusive. PMID- 12554295 TI - Role blurring--does it have implications for staff structure and development? AB - Within recent times medical illustration departments have been changed dramatically by computers and new working methods. For photographers and illustrators alike this has resulted in an overlap or 'blurring' of roles. This paper discusses typical core skills that cause this overlap and looks at the necessity for everyone within the profession to acquire them. PMID- 12554296 TI - Using PowerPoint. AB - PowerPoint is a very flexible program and, for the most part, there is no right and wrong way to do things. Views expressed here are those of the authors and the methods used have been tried and tested. There are three articles in the series. The first article looks at the design issues that need to be considered when preparing a PowerPoint presentation. The second article will concentrate on the 'mechanics' of the program, and the third article will address issues around delivering the presentation. It is hoped that this series will stimulate others to contribute ideas, tips and experiences that can be built into a useful resource. Any contributions should be addressed to the Editor. PMID- 12554298 TI - Gallery. The Leeds Photovisual Awards to the Institute of Medical Illustrators 2002. PMID- 12554297 TI - Making and using visual and audio recordings of patients (May 2002). PMID- 12554301 TI - The Health Sciences and Communications Association (HeSCA) annual conference, Winnipeg, 19-22 June 2002. PMID- 12554299 TI - Joan Fairfax Whiteside ARRC FMAA (1904-2001)--illustrator, portrait painter & medical artist. PMID- 12554309 TI - Defining "early dementia" and monitoring intervention: what measures are useful in family caregiving? AB - Measures of cognition are often used to define and measure the progress of dementia and outcomes of intervention. This paper examines whether measures of psychosocial disability used with those of cognition are more useful than measures of cognition alone, particularly in early dementia. A measure of cognition and two instruments of caregiver burden, used as routine clinical outcome measures of three types of Old Age Psychiatry dementia services, were examined. All cases with dementia in a memory clinic (MC; n = 149), a community mental health service for older people (CMHT; n = 120) and a specialist dementia day hospital (DH; n = 118), in one NHS district were followed up at 12 months. Measures of cognition (MMSE), behaviour, caregiver coping (Problem Checklist; PC) and caregiver mood (Hospital Anxiety and Depression Scale; HAD) were taken at baseline (MC, n = 48; CMHT, n = 113; DH, n = 55) and at follow-up (MC, n = 35; CMHT, n = 34; DH, n = 23). At baseline, all three groups had an average MMSE score of "mild impairment" but measures of behaviour and caregiver burden showed subtle between-group differences. At the 12-month follow-up, cognition remained stable in all groups, but the frequency of day-to-day problems increased and caregiver mood deteriorated in families receiving DH support. The use of psychosocial measures of disability in conjunction with those of cognition, are important in the definition and longitudinal measurement of intervention and support in early dementia. PMID- 12554310 TI - Cognitive rehabilitation as a component of early intervention in Alzheimer's disease: a single case study. AB - Early intervention in Alzheimer's disease (AD) should focus on psychological and social needs as well as the provision of medication. One possible component of early intervention programmes for people with early-stage AD is cognitive rehabilitation aimed at fostering the development of strategies for coping with memory problems. The likely relevance of cognitive rehabilitation in early-stage AD is supported by neuropsychological and experimental learning studies, but further work is required to develop clinically relevant interventions, which can be applied in the real-life setting. This paper presents a single case intervention study in which a 66-year-old man with early-stage AD learned the names of 13 members of his support group using a mnemonic strategy coupled with either expanding rehearsal or repeated presentation, or both, within an errorless learning paradigm. Recall scores improved from a mean of 2.31% at initial baseline to 91.46% following intervention, and gains were largely maintained at follow-up. There was no evidence of any increase in depression, anxiety or caregiver strain during the intervention. The results support the view that cognitive rehabilitation interventions may form a valuable component of comprehensive early intervention programmes for people with AD. PMID- 12554311 TI - An electronic memory aid to support prospective memory in patients in the early stages of Alzheimer's disease: a pilot study. AB - The use of an electronic memory aid (EMA) for patients with mild-to-moderate probable Alzheimer disease is examined in five outpatients aged 58-79 years. The ability to remember to carry out seven tasks at a particular time was evaluated in three experimental conditions: recall without an external memory aid, recall with a written list and recall with support available from an EMA. The use of an EMA significantly improved patients' prospective memory, while the written list and free recall were not useful. Future research that examines the value of using an EMA to help with tasks that are associated with prospective memory with a larger sample of patients within their own home context is suggested. PMID- 12554312 TI - Decision support software for dementia diagnosis and management in primary care: relevance and potential. AB - Dementia, which affects a large and growing number of older people, presents particular challenges to primary care. There is an acknowledged need to develop interventions that address practitioners' needs for information and guidance regarding the diagnosis and management of dementia. This paper examines the potential usefulness and constraints of a Computer Decision Support System (CDSS) to assist practitioners in diagnosing and managing dementia. Questionnaire information was obtained from 97 primary care practitioners regarding their current practice and views on dementia care, priority given to training and familiarity with computer use. Implications of these findings for the relevance and value of CDSS are discussed. The paper is part of a larger ongoing study, the aim of which is the evaluation of three educational interventions for primary care practitioners. PMID- 12554313 TI - Information for dementia patients and their caregivers: what information does a memory clinic pass on, and to whom? AB - This study assessed the information provided in a memory clinic and the patient and caregiver factors that influenced the provision of information. The study was part of a larger cross-sectional study of the diagnosis of dementia and satisfaction with information given. The participants were 51 elderly patients suspected of having dementia and living at home together with their caregivers. The main outcome measures were the checklist of information communicated to the patients and caregivers, patients' behavioural problems, patients' instrumental activities of daily living (IADL) problems, and caregivers' sense of competence. The results indicate that basic information about the diagnosis was given to 86% of patients and 88% of caregivers. Specific information about patients' behaviour and professional care was more often given when patients had more cognitive, behavioural, or IADL problems and caregivers had a lower sense of competence. Providing the majority of patients with information is feasible. These results contribute to an explanation of previous findings indicating that caregivers of patients with cognitive problems not related to dementia were less satisfied with the information they received. Adequate information should not only include issues considered relevant by clinicians but should also be tailored to the information needs of patients and caregivers. PMID- 12554314 TI - The COPE index--a first stage assessment of negative impact, positive value and quality of support of caregiving in informal carers of older people. AB - Data was collected in five countries from informal carers of older people (n = 577) via a common protocol. Carers completed: (1) a 17-item version of the Carers of Older People in Europe (COPE) Index, an assessment of carers' perceptions of their role : (2) a questionnaire on demographic and caregiving circumstances : and (3) three instruments included for the criterion validation of the COPE Index (the General Health Questionnaire, the Hospital Anxiety and Depression Scale, and the World Health Organization Quality of Life-BREF). Principal Component Analysis of the COPE Index was followed by internal consistency analysis of emergent components. Scales derived by summing items loading on the components were analyzed for their association with the criterion measures. Two components, negative impact and positive value, emerged consistently across countries. A third component, quality of support was less consistent across countries. Scales derived from the negative impact and positive value components were internally consistent and significantly associated with the criterion validity measures. These two scales and four items drawn from the quality of support component were retained in the final COPE Index. While further testing is required, the COPE Index has current utility in increasing understanding of the role perceptions of carers of older people. PMID- 12554315 TI - Feasibility of a dyadic intervention for management of osteoarthritis: a pilot study with older patients and their spousal caregivers. AB - This study evaluated a novel intervention for older osteoarthritis (OA) patients and their spousal caregivers that consisted of standard patient education supplemented by information related to effectively managing arthritis as a couple. Twenty-four female OA patients and their husbands were randomly assigned to either an educational intervention that was targeted at both patient and spouse or to a patient education intervention that was targeted at only the patient. Findings revealed that both interventions were evaluated favorably but the couple intervention was better attended than the patient intervention. In addition, patients in the couple intervention experienced greater increased efficacy in managing arthritis pain and other symptoms. The findings of this pilot study point to the utility of a dyadic intervention approach to management of OA in late life. PMID- 12554316 TI - Is blood pressure at 68 an independent predictor of cognitive decline at 81? Results from follow-up study "Men born in 1914", Malmo, Sweden. AB - One hundred and sixty-eight men who were participants in the study "Men born in 1914" had their blood pressure and cognitive function examined when they were 68 years old. They were then re-examined at the age of 81. Decline on five standardized cognitive tests at the age of 81 years was studied in relation to four levels of baseline blood pressure at the age of 68. Results from multivariate analyses suggested that blood pressure, when studied in conjunction with other factors related to cognitive function, contribute to a rather modest extent and not unambiguously to the variability in cognitive decline. When decline on each separate test had been combined into a composite measure, where equal weight had been given to memory, crystallized and fluid abilities, it was shown that hypertension stage 2 (SBP 160-179 mmHg or DBP 100-109 mmHg) was associated with greater levels of overall decline. Hypertension stage 3 (SBP > or =180 mmHg or DBP > or =110 mmHg), on the other hand, was associated with lower levels of overall decline. Depressive symptoms at follow-up had a negative effect on cognitive levels and were discussed as potential confounders of hypertension related cerebral lesions. PMID- 12554324 TI - Intense pulsed light source for treatment of facial telangiectasias. AB - OBJECTIVE: The purpose of this clinical study was to evaluate the effectiveness of the first intense pulsed light source (IPL) with dual mode light filtering for treatment of facial telangiectasias, and to evaluate the incidence of adverse including purpura, pigmentation and scars. MATERIALS AND METHODS: Twenty-four patients with facial telangiectasias were treated between one and four times with a new IPL system. This system differs from previous IPLs by eliminating wavelengths longer than 950 nm, which would otherwise lead to non-specific heating of tissue water. The treatments were performed at one-month intervals. Two months after the last treatment, the clinical effect was evaluated from close up photographs. RESULTS: After one to four IPL treatments (mean: 2.54; SD: 0.96) for facial telangiectasias, 79.2% of the patients obtained a more than 50% reduction in number of vessels, and 37.5% obtained between a 75% and 100% reduction. Moderate erythema and oedema were the only adverse effects of the treatment. No purpura was registered and no long-term adverse effects such as scars or pigmentary disturbances occurred. CONCLUSIONS: An IPL with dual mode filtering is efficient and safe for treatment of facial telangiectasias. PMID- 12554325 TI - Epilation with a long-pulse 1064 nm Nd:YAG laser in facial hirsutism. AB - BACKGROUND: Facial hirsutism and hypertrichosis are common problems and a methods are available to clinicians for the unwanted hair in a large number of patients. Several depilatory laser systems are now available and one of these modalities is the long-pulse Nd:YAG laser. OBJECTIVE: To evaluate the efficacy of the long pulse Nd:YAG laser in removing unwanted facial hair. METHOD: In all, 29 patients completed their treatment course with a long-pulse (4 ms) Nd:YAG (1064 nm line) with fluences between 56 J/cm(2) and 70 J/cm(2). The average reduction in hair density was assessed using hair count on digital photographs at removal 3, 6 and 9 months postoperatively. The hair-free interval was variety of also evaluated. RESULTS: The average reduction of in the hair count (the same diameter as the baseline for the criterion) was 43% at 3 months, 36% at 6 months, and 46% at 9 months. The hair-free interval with laser was two- to sixfold longer than with self-applied methods. No significant complications were observed in the range of skin types treated, even in the darker skin types. CONCLUSION: The long-pulse Nd:YAG laser is an effective and safe method for long lasting hair removal in all skin types. The delay in hair regrowth most likely explains patient satisfaction. PMID- 12554326 TI - Intracutaneous botulinum toxin A versus ablative therapy of Hailey-Hailey disease -a case report. AB - BACKGROUND: Hailey-Hailey disease is an autosomal-dominant blistering disease affecting the intertriginous skin. Dermabrasion and ablative laser treatment are known to be curative. Sweating is a common aggravating factor. Botulinum toxin A (BTXA) has been shown to inhibit sudoriferic nerves. OBJECTIVE: To evaluate whether a treatment with BTXA induces remissions and can compete with ablative therapy. To compare dermabrasion with erbium:YAG laser therapy. METHOD: Case report with side-by-side comparison. We used intracutaneous BTXA on both sides of the submammary region. Four days later a limited area of 25 cm(2) on each side was treated with either dermabrasion or erbium:YAG laser. The follow-up was 12 months. RESULTS: Wound healing was complete within 7 days after erbium:YAG laser and two weeks after dermabrasion. Areas treated with BTXA alone also showed complete remission within two weeks. During a follow-up, no relapse occurred with either treatment. CONCLUSION: BTXA is capable of inducing remissions of Hailey Hailey disease without abrasion for at least 12 months. Among ablative treatments, erbium: YAG laser therapy leads to a more rapid wound closure than dermabrasion, with both causing complete remissions. PMID- 12554327 TI - Pearls from the 10th International Symposium on Cosmetic Laser Surgery, Las Vegas, April 2001. PMID- 12554332 TI - The soul of the knife. The essence, the being of the surgical profession. PMID- 12554333 TI - Origins of surgery in British Colonial America. PMID- 12554334 TI - Use of taurine as antioxidant in resuscitating livers from non-heart-beating donors by gaseous oxygen persufflation. AB - In this study, the importance of antioxidants during oxygenated liver preservation has been investigated. Livers were excised from rats after 60 min of cardiac arrest and stored for 24 h at 4 degrees C in University of Wisconsin solution (UW). Gaseous oxygen was applied to the livers during the storage period via the caval vein after superoxide dismutase (SOD, 600 U/mL), taurine (0.5 mg/mL), or no antioxidant was applied to the graft with the rinse solution. It was shown that oxygen persufflation significantly enhanced the viability of livers during cold preservation only in combination with SOD or taurine, which were both equally effective in reducing lipid peroxidation, enzyme release, and vascular resistance upon postischemic reperfusion. An increase in hepatic bile production was also observed. PMID- 12554335 TI - Systemic endotoxin and gastric mucosal pH are the best parameters to predict lethal outcome in a porcine model of abdominal sepsis according to multivariate analysis. AB - This study was devised to identify sepsis-relevant parameters that early and reliably predict a lethal outcome in intra-abdominal sepsis. In 18 Duroc pigs, peritonitis was induced through standardized gastrotomy. Twelve hours later the defect was oversewn and the abdominal cavity lavaged thoroughly. Sepsis relevant parameters were measured before initiating therapy, and 30 min later animals were extubated and observed for a period of 6 days under adequate analgesia with free access to water and food. All parameters were correlated with survival postoperatively. In the treatment group, 7 out of 18 pigs (39%) died within the observation period. Endotoxin level at 30 min after initiation of therapy [17.9 EU/mL (+/- 12.1) vs. 110.9 EU/mL (+/- 21); p <.001] and Delta pHi [0.015 (+/- 0.011) vs. -0.039 (+/- 0.013); p =.016] were identified as the two parameters with highest predictive power regarding mortality in a multivariate analysis. In conclusion measurement of endotoxin and gastric tonometry should gain wider clinical application in septic patients. PMID- 12554337 TI - Colosigmoid junction: a study of its functional activity with identification of a physiologic sphincter and involvement in reflex actions. AB - The colosigmoid junction (CSJ) marks the termination of the descending colon (DC) and the beginning of the sigmoid colon (SC); it is a fixed area in the retroperitoneum. At this site where two functionally different areas meet, we hypothesized the presence at the CSJ of a physiologic sphincter that regulates the passage of gut contents from the DC to the SC. This hypothesis was investigated for this communication. Eight subjects (mean age 36.6 +/- 4.7 SD years, 6 women) were studied during surgical repair of incisional hernia or laparotomy. The pressure responses of the CSJ to individual distension of the DC and SC were recorded. A balloon-ended tube was introduced per annum to lie in the DC or SC, and the pressure in the DC, CSJ, and SC was measured by saline-perfused catheters. To study whether the CSJ response to individual DC or SC distension was a direct or reflex action, the test was repeated in six of eight patients after separate anesthetization of the DC, CSJ, and SC. The CSJ had a higher pressure than that of the DC or SC; the high-pressure zone measured a mean of 2.1 +/- 0.9 cm. High-volume DC distension effected a significant DC pressure rise (p <.001) and a CSJ pressure decline (p <.05), which lasted a mean of 7.2 +/- 1.2 s. In contrast, the CSJ responded to big volume SC distension by significant pressure elevation (p <.001) which was also momentary. Small volume distension of the DC or SC effected no significant CSJ pressure response (p >.05). The CSJ pressure did not respond to distension of the anesthetized DC or SC. Likewise, the anesthetized CSJ did not react to DC or SC distension. When the test was repeated using saline instead of xylocaine, the CSJ pressure response was similar to that without saline injection. The CSJ is a high pressure zone with a measurable length. It reacts to DC or SC balloon distension by dilatation or narrowing, respectively. These findings presumably denote the existence of a "physiologic sphincter" at the CSJ, which appears to regulate the passage of colonic contents to the SC. We postulate that the CSJ pressure response to DC or SC distension is reflex and mediated through the "colosigmoid reflexes." The role of the colosigmoid sphincter and reflexes in colonic motility disorders remains to be investigated. PMID- 12554336 TI - Disruption of the small-intestine mucosal barrier after intestinal occlusion: a study with light and electron microscopy. AB - It is known that the gut may serve as a reservoir for various microorganisms, which under specific circumstances may intrude into the systemic circulation, causing systemic infections. The aim of the present study was to estimate the "critical time" of disruption of the small-intestine mucosal barrier in conditions of experimentally induced intestinal occlusion, based on the histopathological alterations observed under light and electron microscopy. Forty rabbits underwent small-intestine obstruction through ligation with a nonabsorbable suture. Blood cultures from portal vein and inferior vena cava, as well as cultures from the peritoneal fluid, a hepatic fragment, and a mesenteric lymph node, were obtained before the ligation (0 h). The same cultures were repeated at 4 and 8 h (group A, 20 rabbits) and at 6 and 12 h after the ligation (group B, 20 rabbits). Small-intestine specimens proximal to the occlusion were taken for examination under the optic and electronic microscope in the same time intervals. Five of 20 rabbits of group A died within 4 h and 6 of 20 rabbits of group B died within 6 h after the operation. All anaerobic cultures were negative. All aerobic cultures that became positive developed Escherichia coli colonies. Intestinal epithelium of dead animals was transformed to cuboid with destruction of goblet cells and alteration in secretion of acid polysaccharides. The mucosal appearance of all rabbits that survived 12 hours after ligation was the same. The disruption of the mucosal barrier begins 4 h after complete intestinal occlusion. At 12 h after complete intestinal occlusion, the disruption is total with different degrees of severity. PMID- 12554338 TI - A novel aortic coarctation model for studying hypertension in the pig. AB - We have developed a reproducible renovascular model of hypertension via a controllable, suprarenal aortic coarctation in the pig. This model has many potential applications, including investigation of the effects of acute hypertension in the conscious animal; identification of cardiac and vascular adaptations to chronic hypertension and their reversal; determining the effect of pharmacologic agents or other interventions on hypertension; and furthering our understanding of the implications of chronic hypertension on neurologic function. A totally implantable system was devised by attaching a reinforced silicone vascular occluder to a vascular access port. The occluder was placed around the suprarenal aorta proximal to the diaphragm. Ten pigs were made hypertensive by sequentially inflating the occluder. In six pigs, telemetric monitoring of blood pressure was used to determine when the pigs had reached target pressures (mean arterial blood pressure >150 mm Hg). Four pigs did not have telemetry units placed and blood pressure and heart rate were monitored for 4 weeks by periodically restraining the pigs in a sling. Two pigs reversed their occlusion due to presumed technical errors; the remaining pigs were studied for 4 (n = 5) or 8 (n = 3) weeks and then euthanized. Advantages of this model of aortic coarctation are that the occlusions are performed in awake animals and excessive occlusion of the aorta resulting in neurologic dysfunction or other distress to the animal can be easily corrected by simply withdrawing a small amount of the fluid used for inflation of the occluder. Additionally, removal of the constriction does not require a second surgical procedure. PMID- 12554339 TI - Mesh prosthesis versus triangular flap from the anterior sheath of the abdominal rectus muscle in primary inguinal hernia repair. AB - A study was carried out to compare the use of two different tension-free surgical techniques, for the repair of primary inguinal hernia in cases without complications. The study was carried out on 91 consecutive patients who presented with primary and uncomplicated inguinal hernia. The patients were randomly divided into two groups. Group I consisted of 42 patients whose inguinal hernia was repaired by means of the application of a triangular flap rotated from the anterior sheath of the abdominal rectus muscle and then sutured to the inguinal ligament with non-continuous and nonabsorbable suture 00. Group II consisted of 49 patients whose inguinal hernia was repaired by tension-free hernioplasty, placing a mesh umbrella plug in the deep inguinal orifice and, if necessary, mesh reinforcement to the floor of the inguinal canal (the Gilbert technique). There were no statistically significant differences with regard to age, sex, hernia evolution time, trans- or postoperative complications, or hernia recurrence during an average follow-up time of 3 years. Patients in the flap group required significantly more surgery time and postoperative analgesics. Cost analysis showed that patients receiving a muscle sheath flap spent less money on the actual surgical procedure, given the saving on the purchase of a prosthetic mesh. However, when taking into account the greater need for analgesia and the longer time in the operating theater, there was no difference in overall cost between the two procedures. Inguinal hernia repair using a triangular flap from the anterior sheath of the abdominal rectus muscle is an additional surgical procedure that can be utilized in cases where prosthetic mesh repair is not feasible. PMID- 12554354 TI - Differential regulation of aminopeptidase N (CD13) by transendothelial migration and cytokines on human eosinophils. AB - Aminopeptidase N (CD13) is a cell surface metalloprotease involved in growth regulation, tumor invasion, and down-regulation of regulatory peptides. CD13 expression on eosinophils in bronchoalveolar lavage (BAL) of asthmatics 10 minutes and 18 hours after segmental allergen provocation was significantly increased (+225% to +294%) compared to blood eosinophils. In vitro CD13 expression could be induced on blood eosinophils by transendothelial migration of the cells across interlenkin (IL) 1beta-activated human umbilical cord vein endothelial cells (HUVECs) as well as by the exposure to the cytokines IL-3, IL 5, and granulocyte-macrophage colony-stimulating factor (GM-CSF). The cytokines GM-CSF and IL-5 were significantly less effective in inducing CD13 compared to IL 3. The IL-3-induced expression of CD13 was decreased in the presence of the protein-synthesis inhibitor cycloheximide (-8.8%). Moreover, blocking of CD13 by the protease inhibitors actinonin and bestatin significantly enhanced migration (+40.0% to +80.0%) of eosinophils across HUVEC monolayers. In summary, the data suggest that CD13 is regulated both by the process of transmigration and by the cytokine IL-3. Further, CD13 itself seems to be involved in the process of eosinophil transmigration. aminopeptidase Nendothelial cellseosinophilsinterleukin-3interleukin-5transendothelial migration PMID- 12554356 TI - Methotrexate stimulates lung epithelial cells to release inflammatory cell chemotactic activities. AB - Methotrexate-induced pneumonitis has been reported as an infrequent but potentially serious complication of therapy in a variety of malignant and benign conditions. Because inflammatory cell infiltration is concerned with the development of methotrexate-induced pneumoinitis, and because airway epithelial cells participate in the orchestration of lung inflammation, the authors determined whether methotrexate might stimulate airway epithelial cells (A549 cells) to release neutrophil, monocyte, and eosinophil chemotactic activities (NCA, MCA, and ECA). A549 cells released NCA, MCA, and ECA in a dose- and time dependent manner in response to methotrexate. Partial characterization revealed the heterogeneity of NCA, MCA, and ECA. The release of chemotactic activity was blocked by lipoxygenase inhibitors and cycloheximide. NCA was inhibited by leukotriene (LT) B(4) receptor antagonist, and anti-interleukin (IL)-8 and granulocyte colony-stimulating factor (G-CSF) antibodies. MCA was attenuated by LTB(4) receptor antagonist, and anti-monocyte chemoattractant protein (MCP)-1 and granulocyte-macrophage CSF (GM-CSF) antibodies. ECA was attenuated by LTB(4) receptor antagonist, and anti-IL-8 and GM-CSF antibodies. The release of IL-8, G CSF, MCP-1, GM-CSF, and LTB(4) from A549 cells significantly increased in response to methotrexate. The mRNA expression of IL-8 and MCP-1 was augmented by methotrexate stimulation. These data suggest that type II epithelial cells may modulate inflammatory cell recruitment into the lung by releasing NCA, MCA, and ECA in response to methotrexate. PMID- 12554355 TI - Maternal nicotine exposure during pregnancy and lactation: I. Effect on glycolysis in the lungs of the offspring. AB - We investigated the effect of maternal nicotine exposure during pregnancy and lactation on carbohydrate metabolism in the neonatal lung. Female rats received nicotine (1 mg/kg body weight/day) subcutaneously from day 7 after mating. Control animals received saline. The suckling rats were killed 24 hours after the last dose of nicotine was administered to the mother on postnatal day 14. The lung tissue of 9 rat pups from 3 litters was surgically removed and the in vitro utilization of exogenous glucose (micromol/g wet lung tissue/h) was determined. Lactate production (micromol/g wet lung tissue/h) was also determined to assess glycolytic activity. Maternal nicotine exposure during pregnancy and lactation stimulated glucose turnover by 21.6% (P<.01), but suppressed glycolysis by 24.6% (P<.001) and glycogenolysis by 37.9% (P<.001). Maternal nicotine exposure during gestation and lactation had no effect on the activity of hexokinase (U/g wet lung tissue), but resulted in a lower phosphofructokinase activity (U/g wet lung tissue) in the lungs of the offspring. From the data, it appeared that the inhibition of the flux of glucose through the glycolytic pathway can be attributed to an inhibition of phosphofructokinase. PMID- 12554357 TI - A unique pattern of Tie1 expression in the developing murine lung. AB - The molecular mechanisms responsible for organ-specific differences in vascular development are not well established. Animals lacking the receptor tyrosine kinase Tie1 die of hemorrhage and pulmonary edema. Furthermore, cells lacking Tie1 are excluded from blood vessels of the mature lung. These findings suggest the importance of Tie1 in the pulmonary vasculature. We quantified the organ specific expression of Tie1 during embryonic and postnatal murine development using both quantitative real-time polymerase chain reaction (PCR) and chemiluminescence employing a tie1.lacZ reporter. In the lung, Tie1 expression increases markedly immediately prior to birth and rises further in the newborn animal, a pattern not found in other organs. Furthermore, expression of Tie1 in the lung is also unique by its persistent increase in the adult animal. This unique pattern of Tie1 gene expression in the embryonic and mature lung supports a distinct role for Tie1 in the development and function of the pulmonary vasculature. PMID- 12554358 TI - Microencapsulation of low molecular weight heparin into polymeric particles designed with biodegradable and nonbiodegradable polycationic polymers. AB - Owing to its lack of oral absorption, heparin has to be administered parenterally. However, parental administration has negative aspects such as multiple injections, possible infection, patient inconvenience, and high cost. Now, low molecular weight heparin (LMWH) is taking part in antithrombotic treatment and is proven to confer more advantages than unfractionated heparin. The aim of our present study was to formulate, by the w/o/w emulsification process, LMWH microparticles as potential oral carriers prepared with biodegradable (poly-epsilon-caprolactone and poly-lactic-co-glycolic acid) and nonbiodegradable polycationic polymers (Eudragit RS and RL), used alone or blended. The encapsulation efficiency ranged from 16 to 47% and was highly dependent on the presence of the positively charged polymers. In the same way, a low in vitro LMWH release was observed when Eudragit polymers composed totally or partially the polymeric matrix, compared with biodegradable polymers exhibiting higher LMWH release (40 and 60%). For each formulation, LMWH released from microparticles preserved its biological activity as shown by the antifactor Xa activity. Experiments performed with fluorescein-labeled LMWH showed the drug distribution in microparticles and may give information about the mechanisms controlling LMWH encapsulation and release. PMID- 12554359 TI - Carrier erythrocytes: an overview. AB - Application of erythrocytes, the most abundant cells of the human body with desirable physiologic and morphologic characteristics, in drug delivery has been exploited extensively. These cellular carriers, having remarkable biocompatibility, biodegradability, and life-span in circulation, can be loaded by a wide spectrum of compounds of therapeutic value using different chemically, as well as physically, based methods. Most of the characteristics of the erythrocytes, including shape, membrane fragility, deformability, and hematologic indices undergo some degree of irreversible changes during the loading procedure. The efflux pattern of the encapsulated compounds from the carrier erythrocytes covers a wide range between a relatively rapid release (complete release within a few hours) and no detectable release until the cell lysis. A series of methods have been tested successfully for improvement of in vitro storability of the carrier erythrocytes without any significant changes in cell biology as well as drug delivery efficacy. Carrier erythrocytes have been exploited for several potential applications, including intravenous slow release of therapeutic agents, enzyme therapy, drug targeting to reticuloendothelial system (RES), improvement of oxygen delivery to tissues, and preparation of fused cells. PMID- 12554360 TI - Sodium alginate microspheres of metoprolol tartrate for intranasal systemic delivery: development and evaluation. AB - Bioadhesive sodium alginate microspheres of Metoprolol tartrate (MT) for intranasal systemic delivery were prepared to avoid the first-pass effect, as an alternative therapy to injection, and to obtain improved therapeutic efficacy in the treatment of hypertension and angina pectoris. The microspheres (Ms) were prepared using emulsification--cross-linking method. The formulation variables were drug loading, polymer concentration, cross-linking agent concentration, and cross-linking time. The Ms were evaluated for characteristics, like particle size, incorporation efficiency, swelling ability, in vitro bioadhesion, in vitro drug release, and in vivo pharmacodynamic performance in rabbits against isoprenaline-induced tachycardia. Treatment of in vitro data to different kinetic equations indicated matrix-diffusion controlled drug delivery from sodium alginate Ms. Polymer concentration, cross-linking agent concentration, and cross linking time influenced the drug release profiles significantly. In vivo studies indicated significantly improved therapeutic efficacy of MT from Ms with sustained and controlled inhibition of isoprenaline-induced tachycardia as compared with oral and nasal administration of drug solution. PMID- 12554361 TI - Cell culture studies of a carborane cholesteryl ester with conventional and PEG liposomes. AB - A new cholesterol-carborane conjugate (BCH) has been synthesized as a potential targeting agent for boron neutron capture therapy (BNCT) of cancers. The compound is extremely water insoluble and was formulated in two liposomal formulations to determine if the compound could be adequately taken up by 9L rat glioma cells in cell culture. Several factors potentially affecting the cellular uptake were evaluated, such as concentration of BCH in the incubation medium, incubation time, cell confluence, and the addition of polyethylene glycol (PEG) phospholipids to the liposomal formulation. The studies indicated that the cellular uptakes of BCH in the conventional and PEG liposomal formulations were 49.1 and 45.9 microg boron/g cells, respectively. Therefore, this compound, formulated in both liposomal formulations, delivered sufficient levels of boron to cancer cells in vitro, indicating that BCH is a promising approach for use in BNCT. The uptake appeared to depend upon BCH concentration in the media as well as the confluence of the cells. The greater boron uptake by nonconfluent cells indicated that active growth of cells was a factor in the uptake of this compound. PMID- 12554362 TI - A kinetic model for predicting the release rate of sparingly-water-soluble drugs from a hydrogel-coated polymeric matrix. AB - Medical devices used for on-target drug delivery are often coated with a hydrogel coating for friction-reduction purpose. Thus, the delivery of a sparingly-water soluble drug by such a device must diffuse through a nonerodable hydrogel layer. An empirical rate equation has been derived for such a kinetic model and predicts that the rate of drug release from such a device is directly proportional to the loading of the drug in the polymeric matrix. The validity of this kinetic model was examined by measuring the rate of release of 2,4,4'-trichloro-2' hydroxydiphenyl ether from different hydrogel-coated (ethylene-vinyl acetate) copolymer stents containing a wide range of the drug. The experimentally determined release rates are in reasonably good agreement with those calculated from the empirical rate equation. Bioefficacy test results based on zone-of inhibition test against Escherichia coli are also in good agreement with the release rate and drug-loading data predicted according to the empirical rate equation. PMID- 12554363 TI - Stability of a transdermal salmon calcitonin formulation. AB - This study was designed to monitor the stability of salmon calcitonin during storage conditions, under the electric fields generated during iontophoresis and electroporation, in contact with transdermal glass diffusion cells, and during transport through skin. The formulation in a citrate buffer (pH 4.0) was stable in storage for short-term studies but degraded significantly on extended storage. Albumin was able to minimize adsorption in contact with glass surfaces, and aprotinin was able to minimize proteolytic degradation in contact with skin. The formulation was stable under electric field, but there was a loss due to adsorption if salt bridges were used. PMID- 12554364 TI - Delivery of didanosine from enteric-coated, sustained-release bioadhesive formulation. AB - The aim of our study is to assess the release characteristics, in vitro permeation, and stability of an enteric-coated, bioadhesive, sustained-release formulation of didanosine (ddI). Enteric-coated tablets of ddI, containing Polyox WSRN-303 and Methocel K4M, were prepared using hydroxypropylmethylcellulose phthalate (HPMCP 5.5). The enteric-coated formulation was resistant to dissolution in 0.1 N HCl solution but dissolved within 10 min (+/-2 min) in pH 7.4 phosphate buffered saline. The release profiles were linear with square root time. Stability studies indicate that the formulations were stable at 4 degrees C, room temperature, and 40 degrees C upon storage for 6 months. Polyox WSRN-303 tablets exhibited a higher ddI permeation ratio across live intestinal tissue compared with conventional tablets. Enteric-coated, sustained-release, bioadhesive tablets deliver ddI in small doses and at the same time prevent acid induced degradation and hence hold a potential to improve ddI's oral bioavailability. PMID- 12554370 TI - Microspheres of alginate-chitosan containing isoniazid. AB - Isoniazid was encapsulated into microspheres of alginate-chitosan by means of a complex coacervation method in an emulsion system. Since the encapsulation of isoniazid tends to be limited by its hydrophilic characteristics, this study proposes its microencapsulation by adsorption. The particles were prepared in three steps: (1) preparation of a W/O emulsion; (2) phase separation; and (3) adsorption of the drug. The isolated particles were placed in a solution of the drug under stirring to allow adsorption. The morphology and particle size were analysed by scanning electron microscopy (SEM). The isoniazid content was determined by extraction in 1 M phosphate buffer pH 7.5 under stirring for 4 h. Finally, the samples were filtered and analysed in an UV/VIS spectrophotometer at 260 nm. In vitro release tests were carried out in 0.05 M phosphate buffer pH 7.5. The results showed that microspheres of alginate-chitosan obtained were of spherical shape. The emulsion used for microparticle formation allows the preparation of particles with a narrow size distribution. The adsorption observed is probably of chemical nature, i.e. there is an ionic interaction between the drug and the surface of the particles. PMID- 12554371 TI - Microencapsulation of beta-galactosidase with Eudragit L-100. AB - Microcapsules containing beta-galactosidase (lactase) were prepared by solvent evaporation using the pH sensitive polymer, Eudragit L-100. Formulations were prepared using various polymer-enzyme ratios with total solids content of the internal phase using sucrose stearate as a droplet stabilizer. Particle size distributions were invariant to relative proportion of ingredients but were dependent on stirring conditions. Although sucrose stearate had no effect on particle size distribution, release rate or encapsulation efficiency, its presence at a minimum 2% level was necessary to ensure intact microcapsules. Encapsulation efficiencies were higher for formulations prepared with 15% compared to 10% total solid content. DSC results revealed an interaction between encapsulated Eudragit L-100-enzyme-sucrose stearate vs their physical mixtures. The enzyme activities of the freshly prepared product vs those stored under stressed condition (40 degrees C and 75% RH) were 68 and 40% of their pre processing activity, respectively. In vitro dissolution showed no enzyme release at 1 h in acidic media but 80% of the lactase was released from the microcapsules over 2.5 h in pH 6.8 media, thus establishing the feasibility of lactase microencapsulation to retard enzyme release in an acidic environment and ensuring release at intestinal pH. PMID- 12554372 TI - Comparison of chitosan and gelatin coated microparticles: prepared by hot-melt method. AB - The aim of this study was to compare the performance of microparticles and their release properties after coating by chitosan and gelatin, respectively. All of the poly(epsilon-caprolactone) (PCL) microparticles were prepared by the hot-melt encapsulation method and indomethacin was selected as a model drug to be encapsulated. All of the coated microparticles retained their spherical shape irrespective of the type of coating material, and the particle size of coated microparticles was similar to the uncoated ones. The indomethacin encapsulation efficiency was in the range of 8.65 +/- 0.08 % - 8.81 +/- 0.04% for uncoated microparticles and 8.22 +/- 0.04% - 8.68 +/- 0.08% for coated microparticles. The release of indomethacin from uncoated microparticles followed a two-exponential release profile, where indomethacin was rapidly released within 4 h during the first release phase, after that approximately 20% of the drug was continuously and slowly released for up to 24 h in the second phase. The similar release profile was observed from coated microparticles irrespective of the times of coating and the types of coating material. Both the natural coating materials, chitosan and gelatin, efficiently reduced the initial burst release and the first phase of drug release, but did not alter the second phase of drug release. In other words, chitosan and gelatin could be used to protect the drug on the surface of microparticles from immediately contacting with the release medium and both possessed the same feature in the delay of drug release. PMID- 12554373 TI - Preparation and release characteristics of polymer-coated and blended alginate microspheres. AB - To prevent a rapid drug release from alginate microspheres in simulated intestinal media, alginate microspheres were coated or blended with polymers. Three polymers were selected and evaluated such as HPMC, Eudragit RS 30D and chitosan, as both coating materials and additive polymers for controlling the drug release. This study focused on the release characteristics of polymer-coated and blended alginate microspheres, varying the type of polymer and its concentration. The alginate microspheres were prepared by dropping the mixture of drug and sodium alginate into CaCl(2) solution using a spray-gun. Polymer-coated microspheres were prepared by adding alginate microspheres into polymer solution with mild stirring. Polymer-blended microspheres were prepared by dropping the mixture of drug, sodium alginate and additive polymer with plasticizer into CaCl(2) solution. In vitro release test was carried out to investigate the release profiles in 500 ml of phosphate buffered saline (PBS, pH 7.4). As the amount of polymer in sodium alginate or coating solution increase, the drug release generally decreased. HPMC-blended microspheres swelled but withstood the disintegration, showing an ideal linear release profiles. Chitosan-coated microspheres showed smooth and round surface and extended the release of drug. In comparison with chitosan-coated microspheres, HPMC-blended alginate microspheres can be easily made and used for controlled drug delivery systems due to convenient process and controlled drug release. PMID- 12554374 TI - Compressed biodegradable matrices of spray-dried PLGA microspheres for the modified release of ketoprofen. AB - A spray-drying technique was used to prepare poly(lactide-co-glycolide) (PLGA) drug loaded microspheres. Ketoprofen was chosen as a model NSAID drug. The microspheres were characterized in terms of morphology, drug content and release behaviour. The spray-dried particles were subject to a direct compression process for the preparation of biodegradable matrix tablets. The spray-dried powders were found to have good compaction properties. Tablets were also prepared from a mixture of microspheres and microcrystalline cellulose, mannitol and hydroxypropylmethylcellulose or sodium alginate. The release of ketoprofen in phosphate buffer (pH 7.4) was significantly sustained, indicating the suitability of using tabletted spray-dried PLGA microspheres for controlled drug delivery. The results show that spray-dried PLGA particles have promising properties as direct compression and release controlling excipients in matrix tablets for oral administration. PMID- 12554375 TI - Study of complex coacervation of gelatin with sodium carboxymethyl guar gum: microencapsulation of clove oil and sulphamethoxazole. AB - Complex coacervation of gelatin with sodium carboxymethyl guar gum was studied. The coacervation was studied as a function of pH, colloid composition and concentration. The efficiency of coacervation was followed by measuring viscosity, coacervate yield and turbidity of test solutions. All the measurements suggest that, as the amount of sodium carboxymethyl guar gum (CMGG) in the colloids increases, the pH at which maximum coacervation happens decreases. Effective coacervation could be realized over the pH range of 2.5-4.0 using different compositions. The efficiency of the CMGG/gelatin system to encapsulate oils and solid particles is demonstrated by successful encapsulation of oil of cloves and sulphmethoxazole. PMID- 12554376 TI - Alginate-diltiazem hydrochloride beads: optimization of formulation factors, in vitro and in vivo availability. AB - Alginate beads containing diltiazem hydrochloride (DTZ) were prepared by the ionotropic gelation method. The effects of various factors (alginate concentration, additives type, calcium chloride concentration and curing time) on the efficiency of drug loading were investigated. The formulation containing a mixture of 0.8% methylcellulose (MC) and 4% alginate cured in 2% calcium chloride for 6 h was chosen as the best formula regarding the loading efficiency. The release rate of DTZ from various beads formulations was investigated. The release of drug from alginate beads followed two mechanisms; by diffusion and relaxation of the polymer at pH 1.2, whilst diffusion and erosion are at pH 6.8. The in vitro release of DTZ from MC-alginate beads showed an extended release pattern which was compared with that from commercially available sustained-release (Dilzem SR) and fast release tablets (Dilzem). Thermal analysis revealed that the drug was molecularly dispersed in the beads matrix. Although the release characteristics of DTZ from Dilzem SR and MC-alginate beads were completely different, the bioavailability of DTZ in dogs was comparable as measured by AUC, MRT and relative bioavailability. The absolute bioavailability of MC-alginate beads and Dilzem SR was 88 and 93%, respectively. PMID- 12554377 TI - Encapsulation of chlorothiazide in whey proteins: effects of wall-to-core ratio and cross-linking conditions on microcapsule properties and drug release. AB - A model drug with limited water-solubility, chlorothiazide, was successfully encapsulated in whey protein-based wall systems cross-linked by glutaraldehyde saturated toluene via an organic phase. The effects of drug content of the core in-wall suspension and of cross-linking conditions on core retention and on microcapsule size, structure and core release properties were investigated. Spherical, surface cracks-free microcapsules ranging in diameter from approximately 200-1300 microm were obtained. Particle size distribution of microcapsules was affected by core content and cross-linking conditions. Core retention in microcapsules prepared at different cross-linking conditions and different wall-to-core ratios ranged from 48.9-81%, from 42.2-76.1% and from 37.3 67.2% in large (L), medium-size (M) and small (S) microcapsules, respectively. In all cases, drug crystals were physically entrapped and embedded throughout the cross-linked protein matrix. Core release from the microcapsules into enzyme-free simulated gastric fluid was governed by a diffusion-controlled mechanism and did not involve erosion or softening of the wall matrix. Rate of core release was significantly affected by a combined influence of core content, microcapsule size and cross-linking density. Complete core release from L, M and S microcapsule prepared at different wall-to-core ratios and cross-linking conditions ranged from 28.6-81.2 h, from 16.8-28.6 h and from 7.2-15.9 h, respectively. Results suggested that whey protein-based wall matrix cross-linked by GAST may provide significant opportunities in modulating the release of an encapsulated core with a limited water solubility. PMID- 12554378 TI - Optimizing formulation factors in preparing chitosan microparticles by spray drying method. AB - The chitosan only, chitosan/Pluronic F68, chitosan/gelatin, chitosan/Pluronic F68/gelatin microparticles and betamethasone-loaded chitosan/Pluronic F68/gelatin microparticles were successfully prepared by a spray-drying method. Microparticle characteristics (yield rate, zeta potential, particle size and tap density), loading efficiencies, microparticle morphology and in-vitro release properties were investigated. By properly choosing excipient type, concentration and varying the spray-drying parameters, a high degree of control was achieved over the physical properties of the dry chitosan powders. SEM micrograph shows that the particle sizes of the varied chitosan composed microparticles ranged from 2.12 5.67 microm and the external surfaces appear smooth. Using betamethasone as model drug, the spray-drying is a promising way to produce good spherical and smooth surface microparticles with a narrow particle size range for controlled delivery of betamethasone. The positively charged betamethasone-loaded microparticles entrapped in the chitosan/Pluronic F68/gelatin microparticles with trapping efficiencies up to 94.5%, yield rate 42.5% and mean particle size 5.64 microm varied between 4.32-6.20 microm and tap densities 0.128 g/cm(3). The pH of particle was increased with increasing betamethasone-loaded amount, but both zeta potential and tap density of the particles decreased with increasing betamethasone-loaded amount. The betamethasone release rates from chitosan/Pluronic F68/gelatin microparticles were influenced by the drug/polymer ratio in the manner that an increase in the release% and burst release% was observed when the drug loading was decreased. The in vitro release of betamethasone showed a dose-dependent burst followed by a slower release phase that was proportional to the drug concentration in the concentration range between 14-44%w/w. PMID- 12554379 TI - Formulation and in vitro/in vivo evaluation of terbutaline sulphate incorporated in PLGA (25/75) and L-PLA microspheres. AB - Terbutaline sulphate (TBS) is widely used in the treatment of bronchial asthma, chronic bronchitis and emphysema. Because of its short biological half life and dosing schedule, a long acting TBS formulation is required to improve patient compliance. The objective of this study was to develop a TBS containing biodegradable microsphere formulation. Poly(D,L-lactide-co-glycolide) (PLGA) and poly(L-lactic acid) (L-PLA) were chosen as matrix materials. A solvent evaporation method was used for preparation of microspheres. Surface morphology, particle size distribution and encapsulation efficiency were investigated. In vitro release studies were performed in pH 7.4 phosphate buffer. In vitro distribution of microspheres were studied in the Swiss albino male mice. All microspheres were spherical in shape and had a porous surface with mean diameters of 9-21 microm. The encapsulation efficiency was influenced by the polymer type, but not the molecular weight. About 90% of the initial amount was trapped in PLGA microspheres, and the remainder was on the surface. In the case of L-PLA, 50% of the total drug was associated with the surface of microspheres. The In vitro release pattern was biphasic characterized by an initial burst phase followed by a slower phase. The L-PLA microspheres released approximately 92% of the initial payload in 72 h. On the other hand, TBS release was increased with an increase in the molecular weight of PLGA. Biodistribution of L-PLA microspheres was characterized by an initially high uptake (35%) by the lungs. All these results suggest that L-PLA and PLGA microspheres have the potential to be used for passive lung targeting. PMID- 12554380 TI - Accessibility, usability and universal design--positioning and definition of concepts describing person-environment relationships. AB - PURPOSE: The aim of this paper is to position, define and discuss three concepts crucial for research and practice concerning person-environment relationships, viz. accessibility, usability and universal design. METHODS: Literature review, synthesized with the authors' research and practice experiences. RESULTS: The authors suggest an instrumental, three-step definition to accessibility, highlighting that accessibility comprises a personal as well as a environmental component, and that accessibility must be analysed by an integration of both. Suggesting the introduction of an activity component, accessibility should partly be replaced by the more complex term usability. Universal design is highlighted as a more process-oriented but less stigmatizing concept. CONCLUSION: This paper contributes to the positioning and definition of concepts describing person environment relationships. The definitions suggested challenge current terminology, but can support in developing more efficient research and practice strategies. In order to develop theory for application to societal planning issues, the definition of concepts is a necessary step. PMID- 12554381 TI - Low back pain--a community-based study of care-seeking and therapeutic effectiveness. AB - PURPOSE: To describe care seeking characteristics among low back pain subjects (LBP); to describe the perceived effectiveness of interventions by recipients of care for LBP; and, to discuss the gap between LBP management guidelines and actual practice in primary care. METHODS: The population consisted of 555 subjects who were recruited from a cross sectional survey of all inhabitants of a small town in Israel. They reported 1 month LBP and were further probed regarding the characteristics of their pain and pattern of care. Data collection was performed by a self-administered questionnaire. RESULTS: Some form of care was sought by 78.7% of the participants. More than half (58%) visited a physician, 34.4% were prescribed drug therapy, 37.1% attended physical therapy and 28.6% turned to alternative medicine. Participants with severe pain were more likely to seek care. That was demonstrated by all pain measures (e.g. high Roland and Morris disability scores (8.2+/-5.6 vs 4.4+/-4.1), high symptoms frequency (8.9+/ 4.1 vs 7.5+/-3.6) and high pain severity scores (5.6+/-2.2 vs 4.5+/-1.7)). The intervention perceived to be most effective was alternative medicine (graded as 'effective' or 'very effective' by 62.8%), followed by therapeutic exercise programmes (56.9%), bed rest (51.9%) and physical therapy (46.8%). CONCLUSIONS: Individuals reporting severe LBP were more likely to seek care. The treatment utilities perceived to be most effective were alternative medicine and therapeutic exercises, whereas the least effective were physicians' office visits. A gap was found between clinical guidelines and physicians referrals for other therapeutic interventions. PMID- 12554382 TI - Treatment of patients with postpolio syndrome in a warm climate. AB - PURPOSE: Treatment in warm climate of various patient groups including patients with postpolio syndrome is controversial. METHOD: Eighty-eight patients with postpolio syndrome (61 women) were recruited, stratified according to sex, age (above/below 60 years old) and use/not use of electrical wheelchair, and randomized to three groups. Group 1 (n=30) underwent treatment in a rehabilitation centre in Tenerife for four weeks in November/December 1999. Group 2 (n=29) were treated in two similar centres in Norway for the same period of time, while Group 3 (n=29), the control group, followed their ordinary health care programme. All patients were tested at the start of study, and 3 and 6 months later, including physical tests and several questionnaire and qualitative interviews. Patients in Group 1 and 2 were also tested after the rehabilitation period. RESULTS: Group 1 and 2 improved significantly both in physical tests and subjective ratings. The positive effects in Group 1 tended to exceed the positive effects in Group 2, and the effects lasted longer. Six minutes walking distance in the two groups was 347 m and 316 m, respectively, before the treatment period, 429 m and 362 m immediately after, and 431 m and 356 m 3 months later. Subjective rating of pain (VAS-scale) was 42 and 43, respectively, before treatment, 17 and 31 immediately after, and 28 and 44 3-months later. In the control group, only minor changes were found. CONCLUSIONS: The study seems to document a positive effect of treatment of patients with postpolio syndrome in warm climate. PMID- 12554383 TI - Understanding the consequences of access barriers to health care: experiences of adults with disabilities. AB - PURPOSE: The study explores: (1) the scope and nature of the consequences that adults with disabilities perceive as the result of inappropriate access to health care services; (2) the variability of these consequences by demographic attributes such as disability type, gender, and health insurance type; and (3) the inter-relatedness and multidimensionality of these consequences. METHODS: Qualitative, semi-structured, in-depth interviews were administered over the telephone to 30 participants with spinal cord injury, cerebral palsy, or multiple sclerosis as part of a nation-wide study on access and utilisation in the USA. Interviews were transcribed and coded for analysis using the qualitative analysis program, NVivo. RESULTS: Consequences were grouped into one of five categories: social, psychological, physical, economic and independence issues. Responses differed slightly with regard to disability type, gender and health insurance type. There was substantial overlap among consequence categories. For most respondents, negative consequences were not limited to just one area--frequently, one consequence triggered others. CONCLUSIONS: Health insurers and providers need a better understanding of the multiple consequences of access barriers. Based on this knowledge, detrimental and costly effects of inappropriate service delivery could be more effectively prevented. Implications for health care services and policy are discussed. PMID- 12554384 TI - Variables related to job satisfaction among professional care providers working in brain injury rehabilitation. AB - PURPOSE: To examine variables that contribute to work satisfaction among rehabilitation professionals involved in brain injury rehabilitation. METHOD: One hundred and thirty-three respondents completed a job satisfaction questionnaire and qualitative questions regarding sources of stress and satisfaction in working with individuals with acquired brain injury (ABI). RESULTS: Factor analysis yielded four factors relating to intrinsic satisfaction, perceived importance to the organization, organizational support and training support. Although intrinsic satisfaction was significantly related to the three other factors, it was best predicted by training support, particularly training that addressed the management of intrusive and/or aggressive behaviours. Qualitative correlates of both work stress and satisfaction were also identified with respect to personal issues, client-related issues, and organizationally-related issues. CONCLUSIONS: Satisfaction with work in ABI rehabilitation is derived from multiple sources, but it appears to be largely dependent on feeling important to, and valued by, the organization and on the level of support and training provided in the workplace. PMID- 12554385 TI - Self-reported efficacy of an all in-the-ear-canal prosthetic device to inhibit stuttering during one hundred hours of university teaching: an autobiographical clinical commentary. AB - This manuscript outlines the lifelong battle with severe stuttering and describes a new modality of effective amelioration of the disorder from the standpoint of a university professor and researcher in the field of stuttering. Childhood reactions to stuttering are discussed, along with the educational and vocational impact of stuttering. Ongoing therapy was received throughout the formative years and into adulthood, emphasizing reduced rates of speech. The use of Delayed Auditory Feedback (DAF) was found to induce fluent speech, but was considered only as a tool for decreasing the speech rate to achieve fluency. When fluency under DAF was discovered to be possible at faster speech rates, the possibility that the use of DAF and other forms of altered auditory feedback could themselves have an inhibitory effect on stuttering, without concomitant rate reduction was investigated. An In-The-Canal (ITC) fluency-enhancing device was used that provided DAF and Frequency Altered Feedback (FAF) to produce more fluent speech. After 10 months of use, the author was relatively free from stuttering. Speech was natural sounding, relatively spontaneous and unlaboured with an absence of fear. However, further testing (that is currently underway at various centres) is necessary before generalizations can be made. PMID- 12554387 TI - Orthopaedic sequelae in neurologically recovered obstetrical brachial plexus injury. Case study and literature review. AB - PURPOSE: Description of shoulder sequelae in obstetrical brachial plexus injury (OBPI) patients who had spontaneous functional recovery, in the context of historical and current conservative methods of treatment. METHOD: Case study of a baby with serious complications, followed by a review of the literature from 1900 until 2001 about conservative treatment of OBPI with respect to the prevention of shoulder complications. RESULTS: Description of contractures and bony deformities did not show important discrepancies over time, other than more detailed images because of new technical possibilities. There is no agreement on the explanation of the development of these deformities. Secondary changes caused by muscular imbalance and longstanding contracture are recognized by all authors. A primary osteoarticular lesion was recognized as a possible cause in the beginning of the twentieth century, then forgotten for a long time and only in the 1980s had gained interest again. The main change in treatment concerns the use of arm braces. This was strongly recommended in the first half of the twentieth century, then advised against and is at this moment not anymore mentioned. CONCLUSIONS: There is no consensus on the cause of contractures and bony deformities in children with OBPI. Conservative methods of treatment have changed over the years, without research on the outcome of these treatment changes. PMID- 12554388 TI - Traumatic brain injury: classification of initial severity and determination of functional outcome. AB - PURPOSE: The aim of the present manuscript is to review current methods for classifying initial severity and final outcome in traumatic brain injury (TBI) and to suggest a direction and form of further research. METHOD: The literature on valid and reliable measurements used in TBI-research for classifying initial severity and final outcome was reviewed. RESULTS: Classifying initial severity in patients with head injury according to clinical condition or CT-parameters is valid. Classifying outcome according to measurement tools of disability showed adequate validity and reliability. CONCLUSIONS: Future research in TBI outcome, particularly in rehabilitation medicine, should focus on determinants of outcome, identifying those patients who will have the greatest chance of benefiting from intensive rehabilitation programmes. More research is needed to determine the long-term functional outcome in TBI, the long-term socio-economic costs, and the influence of behavioural problems on family cohesion. Finally, validation of outcome measures is required in the TBI-population; the relative value of various outcome measures needs to be determined, and the usefulness and applicability of measures for health related quality of life in TBI should be established. PMID- 12554389 TI - Mobility of wheelchair users: a proposed performance assessment framework. AB - PURPOSE: Few standardized outcome measures used in the clinical assessment of wheelchair user performance assess or document all the factors that influence mobility. This article presents a conceptual framework that suggests a more all encompassing view of wheelchair mobility performance assessment. METHOD: After determining, through a survey of the literature, which factors influence wheelchair mobility, a conceptual framework was proposed. Outcome measures of wheelchair mobility were then surveyed. An analysis of these assessment tools allowed factors to be determined that were considered in the clinical assessment of wheelchair mobility performance. A comparison of these led to observations being drawn, which could allow the development of a valid and reliable standardized outcome measure to be used in clinics. RESULTS AND CONCLUSION: Future improvements to the clinical assessment of wheelchair mobility performance should focus on developing an assessment tool considering all categories of factors influencing wheelchair mobility. These categories are the user's profile, the wheelchair, the environment, the daily activities and social roles and the assessment and training received. A controlled-environment outcome measure using a standardized obstacle course with fundamental situations would allow rehabilitation therapists to evaluate clients for the selection of a wheelchair and to document their progress during training. This tool should make it possible to extrapolate the results and thereby apply them to daily activities and social roles. PMID- 12554390 TI - Awareness of disabilities in stroke rehabilitation--a clinical trial. AB - PURPOSE: To investigate the frequency of unawareness of disabilities after stroke during the rehabilitation stage, the relationship of unawareness with neuroanatomical variables, and the impact of unawareness on functional outcomes. METHOD: Sixty consecutive patients (36 with right, 24 with left hemisphere damage) admitted to rehabilitation hospital with a first, single, unilateral stroke were evaluated at admission, discharge and at 1-year post onset of stroke. Unawareness of disabilities was operationally defined as the discrepancy between therapist and patient's rating on the motor scale of the functional independence measure (FIM). Functional outcomes included FIM, instrumental activities of daily living (IADL) scale, activity card sort (ACS) and safety rating scale. RESULTS: Unawareness of disabilities was found in 44/60 patients at admission and 24/57 at discharge. There was no significant difference between the hemisphere groups in the frequency of unawareness at both times. Discharge unawareness in the right hemisphere group was significantly associated with lesions in the frontal and temporal lobes, and with lesion size. Unawareness in the left hemisphere damaged group was not associated with any neuroanatomical variables. A negative impact of unawareness at admission on functional outcomes was not found, but it was found that unawareness at discharge was a negative predictor of activity level (ACS score) at follow up, after controlling for the severity of initial disability level. CONCLUSIONS: Unawareness of disabilities is a significant issue in stroke rehabilitation. Unawareness that persists to discharge from rehabilitation correlates with neuroanatomical variables in right hemisphere damaged patients, and is a negative predictor for some rehabilitation outcomes at follow-up. PMID- 12554391 TI - A comparison of four functional tests in discriminating fallers from non-fallers in older people. AB - PURPOSE: Which functional tests on mobility and balance can better screen older people at risk of falls is unclear. This study aims to compare the Berg Balance Scale (BBS), Tinetti Mobility Score (TMS), Elderly Mobility Scale (EMS) and Timed Up and Go test (TUG) in discriminating fallers from non-fallers in older people. METHOD: This was a case-control study involving one rater who conducted a mobility and balance assessment on subjects using the four functional tests in random sequence. Subjects recruited included 17 and 22 older people with a history of single and multiple falls respectively from a public Falls Clinic, and 39 community-dwellers without fall history and whose age, sex and BMI matched those of the fallers. All subjects underwent the mobility and balance assessment within one day. RESULTS: Single fallers performed better than multiple fallers in all four functional tests but were worse than non-fallers in the BBS, TMS and TUG. The BBS demonstrated the best discriminating ability, with high sensitivity and specificity. The BBS item 'pick up an object from the floor' was the best at screening fallers. CONCLUSION: BBS was the most powerful functional test of the four in discriminating fallers from non-faller. PMID- 12554392 TI - Support for children following traumatic brain injury: the views of educational psychologists. AB - PURPOSE: For many years, neuroscientific research appeared to support the view that the young brain was more resilient and better able to withstand damage than the mature brain. More recent neuroscience research has questioned this view. Nevertheless, the view that 'younger is better' continues to be accepted by many medical practitioners. The present study sought to establish whether the training and experience of educational psychologists has made them aware that young children are particularly vulnerable to long-term effects of brain injury. METHOD: Educational psychologists were asked to estimate the need for additional social/family and educational support, and how long such additional support would be needed, in four fictitious but clinically-based case studies which differed only in the reported age of the child/adolescent at the time of the injury. RESULTS: Educational psychologists consistently rated young children with brain injuries as needing more additional social/family support for longer than adolescents. CONCLUSIONS: These results suggest that there is an appreciation of the additional needs of young children who have sustained brain injuries within our educational system. PMID- 12554394 TI - Festschrift issue for Gavriel Salvendy. PMID- 12554395 TI - Do ergonomics improvements increase computer workers' productivity?: an intervention study in a call centre. AB - This paper examines whether improving physical ergonomics working conditions affects worker productivity in a call centre with computer-intensive work. A field study was conducted at a catalogue retail service organization to explore the impact of ergonomics improvements on worker production. There were three levels of ergonomics interventions, each adding incrementally to the previous one. The first level was ergonomics training for all computer users accompanied by workstation ergonomics analysis leading to specific customized adjustments to better fit each worker (Group C). The second level added specific workstation accessories to improve the worker fit if the ergonomics analysis indicated a need for them (Group B). The third level met Group B requirements plus an improved chair (Group A). Productivity data was gathered from 72 volunteer participants who received ergonomics improvements to their workstations and 370 control subjects working in the same departments. Daily company records of production outputs for each worker were taken before ergonomics intervention (baseline) and 12 months after ergonomics intervention. Productivity improvement from baseline to 12 months post-intervention was examined across all ergonomics conditions combined, and also compared to the control group. The findings showed that worker performance increased for 50% of the ergonomics improvement participants and decreased for 50%. Overall, there was a 4.87% output increase for the ergonomics improvement group as compared to a 3.46% output decrease for the control group. The level of productivity increase varied by the type of the ergonomics improvements with Group C showing the best improvement (9.43%). Even though the average production improved, caution must be used in interpreting the findings since the ergonomics interventions were not successful for one-half of the participants. PMID- 12554396 TI - Development of error-compensating UI for autonomous production cells. AB - This contribution deals with the impact of human error on the overall system reliability in flexible manufacturing systems (FMS). Autonomous production cells are used to illustrate an error-compensating system design on the basis of Sheridan's (1997) paradigm of supervisory control. In order to specify human errors and their effects in terms of system disturbances, a taxonomy of system disturbances is recommended. This taxonomic approach was derived by a value benefit analysis and is based on HEDOMS (Human Error and Disturbance Occurrence in Manufacturing Systems) with slight modifications and Reason's GEMS (Generic Error Modelling System). The taxonomy is used for data acquisition. Next, a risk priority equivalent to FMEA (Failure Mode and Effect Analysis) is introduced to structure the data according to their relevance. Then, Vicente's and Rasmussen's guidelines (1987) for an ecological interface design are related to the paradigm of supervisory control. On the basis of these guidelines four case studies are presented to show their successful applicability for interface design in FMS. PMID- 12554397 TI - Virtual prototyping and testing of in-vehicle interfaces. AB - Electronic innovations that are slowly but surely changing the very nature of driving need to be tested before being introduced to the market. To meet this need a system for integrated virtual prototyping and testing has been developed. Functional virtual prototypes of various traffic systems, such as driver assistance, driver information, and multimedia systems can now be easily tested in a driving simulator by a rapid prototyping approach. The system has been applied in recent R&D projects. PMID- 12554398 TI - Interactive decision support system to predict print quality. AB - Customers using printers occasionally experience problems such as fuzzy images, bands, or streaks. The customer may call or otherwise contact the manufacturer, who attempts to diagnose the problem based on the customer's description of the problem. This study evaluated Bayesian inference as a tool for identifying or diagnosing 16 different types of print defects from such descriptions. The Bayesian model was trained using 1701 narrative descriptions of print defects obtained from 60 subjects with varying technical backgrounds. The Bayesian model was then implemented as an interactive decision support system, which was used by eight 'agents' to diagnose print defects reported by 16 'customers' in a simulated call centre. The 'agents' and 'customers' in the simulated call centre were all students at Purdue University. Each customer made eight telephone calls, resulting in a total of 128 telephone calls in which the customer reported defects to the agents. The results showed that the Bayesian model closely fitted the data in the training set of narratives. Overall, the model correctly predicted the actual defect category with its top prediction 70% of the time. The actual defect was in the top five predictions 94% of the time. The model in the simulated call centre performed nearly as well for the test subjects. The top prediction was correct 50% of the time, and the defect was one of the top five predictions 80% of the time. Agent accuracy in diagnosing the problem improved when using the tool. These results demonstrated that the Bayesian system learned enough from the existing narratives to accurately classify print defect categories. PMID- 12554399 TI - Multimodal feedback: an assessment of performance and mental workload. AB - Multimodal interfaces offer great potential to humanize interactions with computers by employing a multitude of perceptual channels. This paper reports on a novel multimodal interface using auditory, haptic and visual feedback in a direct manipulation task to establish new recommendations for multimodal feedback, in particular uni-, bi- and trimodal feedback. A close examination of combinations of uni-, bi- and trimodal feedback is necessary to determine which enhances performance without increasing workload. Thirty-two participants were asked to complete a task consisting of a series of 'drag-and-drops' while the type of feedback was manipulated. Each participant was exposed to three unimodal feedback conditions, three bimodal feedback conditions and one trimodal feedback condition that used auditory, visual and haptic feedback alone, and in combination. Performance under the different conditions was assessed with measures of trial completion time, target highlight time and a self-reported workload assessment captured by the NASA Task Load Index (NASA-TLX). The findings suggest that certain types of bimodal feedback can enhance performance while lowering self-perceived mental demand. PMID- 12554400 TI - Multiple metaphor environments: designing for diversity. AB - This paper advances a proposition for the engineering of interactive computer based environments capable of exhibiting alternative interactive embodiments to cope with diversity in users, interaction platforms and usage contexts. Such systems are referred to as Multiple Metaphor Environments (MME). The theoretical underpinnings of an MME rely on a conception of HCI design as mapping functions in a machine-oriented language (target domain) to symbols in a user-oriented language (source domain), and vice versa. Such a conception, which is rooted in developments in communication theory and the philosophy of language, constitutes the baseline for formulating a proposal for the design of MME. The proposal comprises a set of engineering principles, process-oriented guidelines and design techniques intended to facilitate a detailed account of how interactive systems could be designed to cope with diversity. To aid the articulation of the various properties of MME, we refer to concrete case studies that provide exemplars of novel insights and promising design practices towards the specification of MME. PMID- 12554401 TI - Effects of training and experience on perception of hazard and risk. AB - This study was designed to show how those proficient at a machining task, where proficiency was gained through both training and past experience, influence the perceived hazard and risk when observing Computer Numerical Control (CNC) machining. The study was also designed to determine whether the impact of the visual and auditory cues can be isolated during diagnosis of the hazard. In a study that included 40 participants, results show that trained observers can perceive more correctly a hazardous condition which is different from what one would predict based on psychophysics. The results also show that trained participants relied strongly on the auditory cues to diagnose the potential hazard and risk correctly whereas the untrained participants who had access to both auditory and visual senses perceived the potential hazard more incorrectly than those trained and more similarly to a traditional Stevens' psychophysical curve. Untrained subjects who had only auditory input had the most difficulty in distinguishing differences in the perception of hazard. This is important considering recent dialogue about whether there is a need to train for tasks that will be automated. The idea of training, especially for automated tasks, is important in allowing better recognition of hazard and risk in unusual circumstances. Additional research may help to improve the diagnosis of hazard and risk and may enable generalization of the results to other training scenarios in the manufacturing and services industries. PMID- 12554402 TI - Perspectives on computer aiding in cognitive work domains: toward predictions of effectiveness and use. AB - A two-stage process is proposed for evaluating computer aiding in cognitive work domains. The first stage relies on a taxonomic framework that facilitates assessments regarding the relative strengths and weaknesses of humans and computers for different computer-aiding schemes. The second stage introduces the construct of tension, which addresses the potential implications of the interplay between computer aiding constraints and human tendencies. This approach is intended to provide designers, analysts and managers with greater insight into the nature of computer aiding and ultimately to enhance their ability to predict its effectiveness as well as its prospects for being used. PMID- 12554403 TI - Work design and analysis for space-based manufacturing: a case analysis of initial design issues. AB - The purpose of the present research was to investigate the nature of potential manufacturing tasks humans may execute in a space environment. The success of space-based manufacturing (SBM) is suggested to be a precursor to permanent human presence in space. A working hypothesis for this study was that human work in the SBM environment would be substantially different from terrestrial manufacturing work. To investigate this hypothesis, a case analysis approach was developed that employed a function allocation and task analysis of a representative SBM process: the production of tailored industrial crystals. This research approach was chosen as the current state of engineering design for SBM is in the conceptual and early flow sheeting phases of a system life cycle. Results of the task analysis and function allocation process suggest response to corrective maintenance functions and to abnormal system conditions should be allocated to humans as opposed to automation. These results are discussed in relation to human factors engineering challenges associated with long-duration human presence in an SBM environment. PMID- 12554404 TI - What users want in e-commerce design: effects of age, education and income. AB - Preferences for certain characteristics of an online shopping experience may be related to demographic data. This paper discusses the characteristics of that experience, demographic data and preferences by demographic group. The results of an online survey of 488 individuals in the United States indicate that respondents are generally satisfied with their online shopping experiences, with security, information quality and information quantity ranking first in importance overall. The sensory impact of a site ranked last overall of the seven characteristics measured. Preferences for these characteristics in e-commerce sites were differentiated by age, education and income. The sensory impact of sites became less important as respondents increased in age, income or education. As the income of respondents increased, the importance of the reputation of the vendor rose. Web site designers may incorporate these findings into the design of e-commerce sites in an attempt to increase the shopping satisfaction of their users. Results from the customer relationship management portion of the survey suggest that current push technologies and site personalization are not an effective means of achieving user satisfaction. PMID- 12554405 TI - Naive and experienced judgments of stimulus-response compatibility: implications for interface design. AB - Many design guidelines encourage maintaining stimulus-response compatibility whenever possible. Payne found that naive judgments for different stimulus response (S-R) mappings were not very accurate, and suggested that designers may not be able to predict whether a particular display-control configuration will lead to better performance than another. Three experiments were conducted to determine whether naive judgments for two-choice tasks in which stimuli and responses involve left-right spatial information are sensitive to (a) the influence of S-R mode relations and (b) pure versus mixed presentation of compatible and incompatible mappings. Initial performance judgments for these conditions were not very accurate, nor were those for four-choice tasks of the type studied by Payne, but subjects' estimates of performance improved with relatively little experience using the different S-R configurations. PMID- 12554406 TI - A data mining technique for discovering distinct patterns of hand signs: implications in user training and computer interface design. AB - Hand signs are considered as one of the important ways to enter information into computers for certain tasks. Computers receive sensor data of hand signs for recognition. When using hand signs as computer inputs, we need to (1) train computer users in the sign language so that their hand signs can be easily recognized by computers, and (2) design the computer interface to avoid the use of confusing signs for improving user input performance and user satisfaction. For user training and computer interface design, it is important to have a knowledge of which signs can be easily recognized by computers and which signs are not distinguishable by computers. This paper presents a data mining technique to discover distinct patterns of hand signs from sensor data. Based on these patterns, we derive a group of indistinguishable signs by computers. Such information can in turn assist in user training and computer interface design. PMID- 12554407 TI - Identification of metaphors for virtual environment training systems. AB - The objective of this effort was to develop potential metaphors for assisting wayfinding and navigation in current virtual environment (VE) training systems. Although VE purports a number of advantages over traditional, full-scale simulator training devices (deployability, footprint, cost, maintainability, scalability, networking), little design guidance exists beyond individual instantiations with specific platforms. A review of metaphors commonly incorporated into human-computer interactive systems indicated that existing metaphors have largely been used as orientation aids, mainly in the form of guided navigational assistance, with some position guidance. Advanced metaphor design concepts were identified that would not only provide trainees with a useful orienting framework but also enhance visual access and help differentiate an environment. The effectiveness of these concepts to aid navigation and wayfinding in VEs must be empirically validated. PMID- 12554408 TI - The development of a theoretical framework and design tool for process usability assessment. AB - The purpose of usability engineering is to facilitate the deployment of new products by decreasing development costs and improving the quality of systems. This paper will discuss the development and delivery of a unique, theoretically based software tool that provides engineers and designers with easy access to the most recent advances in human-machine interface design. This research combines several theoretical views of the interaction process into a hybrid model. Based on this model, a software tool was produced that allows engineers to model the human interaction process within their design. The system then provides feedback on the interaction process through items such as: the amount of mental effort required by a user, the degree to which the system conforms to human capabilities, the expected time to complete the interaction, where potential human error may occur, as well as potential misunderstandings or points of confusion to the users. The designer may then use this information to improve the design of the system. Validation of this technique indicates that the hybrid model produces accurate predictions of usability attributes and that the technique transfers from the laboratory to the real world. PMID- 12554409 TI - Culture differences in design of portal sites. AB - Two experiments were carried out to examine the effects of cultural differences between the Chinese and the US people on the perceived usability and search performance of World Wide Web (WWW) portal sites. Chinese users in Taiwan and US users in Chicago were recruited to perform searching tasks on two versions of Yahoo! portal site: the standard Yahoo! and Yahoo! Chinese. The layout of Yahoo! Chinese is the same as the layout of Yahoo!, and categories on Yahoo! Chinese have been translated from its US counterpart. A special browser was programmed to record all the keystroke data and participants were asked to fill out a satisfaction questionnaire after finishing the tasks. Significant differences of satisfaction and steps to perform some tasks were found between the two groups. The experiment results also provided more detailed insights into the cultural differences between the Chinese and the US users. PMID- 12554410 TI - Internationalization and localization: evaluating and testing a Website for Asian users. AB - The objective of this study was to combine internationalization and localization of Websites and improvement of Website usability with user-centred design methods. This study designed for internationalization and localization of Websites for Asian users, and implemented usability engineering into every phase of Website usability testing, based on the internationalization and localization perspectives of the honeywell.com/your home Website. The first step was to develop the usage scenarios. Three Asian usability specialists carried out one heuristic evaluation session for the current honeywell.com/your home Website. The usability problems were analysed and possible solutions to these problems were discussed. In the next phase, cluster analysis was utilized to test current information architecture. The results provided options for future information architecture development for this Website. Finally, a performance measurement test was conducted to investigate the performance for Asian users. Based on the results, suggestions for improving the Website usability from the localization perspective were provided. The results demonstrate the user-centred design (UCD) approach and stress international and local issues in Website development to Website designers. PMID- 12554411 TI - Design and evaluation of computerized operating procedures in nuclear power plants. AB - A small-scale virtual system has been developed in this study to enhance operators' understanding and operating performance. For this, a computerized graphical interface based on Dynamic Work Causality Equation (DWCE) has been designed to transform the operating procedure into a flowchart. Furthermore, the Programmable Logic Controller (PLC) was installed to connect the signboard (proposed system) with the computerized graphical interface. An experiment was conducted to verify the effect of computerized graphic interface, indicating that the computerized system significantly decreases learning time and improves operational performance. PMID- 12554412 TI - A neuro-fuzzy model for estimating electromyographical activity of trunk muscles due to manual lifting. AB - The main objective of this study was to develop a hybrid neuro-fuzzy system for estimating the magnitude of EMG responses of 10 trunk muscles based on two lifting task variables (trunk velocity and trunk moment) as model inputs. The input and output variables were represented using the fuzzy membership functions. The initial fuzzy rules were generated by the neural network using true EMG data. Two different laboratory-derived EMG data sets were used for model development and validation, respectively. The mean absolute error (MAE) between the actual and model-estimated normalized EMG values was calculated. Across all muscles, the average value of MAE was 8.43% (SD=2.87%) of the normalized EMG data. The larger absolute errors occurred in the left side of the trunk, which exhibited higher levels of muscular activity. Overall, the developed model was capable of estimating the normalized EMG values with average value of the mean absolute differences of 6.4%. It was hypothesized that model performance could be improved by increasing the number of inputs, including additional task variables as well as the subjects' characteristics. PMID- 12554413 TI - Ecological models of human performance based on affordance, emotion and intuition. AB - This paper proposes a complementary approach to Rasmussen's taxonomy of the human skill-, rule-, and knowledge-based performance models by combining the ecological concept of affordances with the neural concepts of human emotion and intuition. The classical cognitive engineering framework is extended through the neuro ecological approach, including personal human attributes important in exercising control over the work environment. The proposed affordance-, emotion-, and intuition-based models correspond to the three types of human performance, namely: learning, adaptive and tuning control, respectively. The new framework is not a predictive model of the operator behaviour, but rather it describes the processes of neuro-ecological control of the human environment. PMID- 12554422 TI - From the editor--enhancing family coping with depression. PMID- 12554423 TI - Formal and informal social networks of women with serious mental illness. AB - The case study method was used to describe formal and informal social networks of four women with serious mental illness and to capture what happens in network interactions. Supportive roles were identified, with significant overlap between roles performed by formal and informal network members. In contrast to many studies, this study's participants all showed reciprocity in relationships. Network relationships, while supportive, often were conflicted. Themes related to stability of relationships were identified. Practice recommendations include recognition of each person's "context" apart from his or her illness and the importance of capitalizing on positive factors within the person's life context. Further research is needed to identify confirming and contradictory findings. PMID- 12554424 TI - Recovery from depression associated with Guillain Barre syndrome. AB - Guillain Barre Syndrome is a rare disease, affecting primarily, but not solely, the peripheral nervous system. Because it is rare, many physicians, nurses, and health care practitioners see few cases in their careers. A case study, in which the patient/author was not diagnosed for approximately a year, is interesting because of associated depression. Recovery from depression is not simple. The steps achieved by the author took a great deal of effort. However, mental health nurses could play a more active role in raising questions for patients without a diagnosis or with evidence of mental confusion, and in assuring empathetic regard. PMID- 12554425 TI - Resilience in sheltered battered women. AB - While many battered women report physical and psychological distress, others are able to respond to adverse sequelae with less severe outcomes. The findings of this study indicate that resilience assessed by the Resilience Scale was significantly and inversely correlated with three global measures and five subscales of the Symptom Checklist-90R. Further study of battered women's resilience will allow for greater appreciation of their strengths as well as expand understanding of human responses to trauma. PMID- 12554426 TI - Psychosocial club members' characteristics and their readiness to participate in a support group. AB - Among persons with severe mental illness, more than half identify problems with loneliness and social isolation. This study examines the demographic characteristics of 34 psychosocial club members and their readiness to participate in a nurse-facilitated support group as a potential means of increasing affiliation. Readiness is represented by the presence of Stone's postulated deterrents to group participation by persons with severe mental illness--unmet survival needs, crises, disinterest in increasing affiliation, satisfaction with life as is, and unwillingness to share information about themselves and their problems. Relationships between these variables and willingness to join, and later to participate, in a group were examined. Despite impaired social/vocational functioning and high levels of potential deterrents, two-thirds of the participants indicated a willingness to join and one-third actually did participate when groups subsequently were offered. Level of social support was positively correlated with both willingness to join and actual participation. That those scoring low on perceived social support were less willing to join a support group suggests that nurses must actively encourage and facilitate participation. PMID- 12554427 TI - Families living with severe mental illness: a literature review. AB - Schizophrenia is a severe mental illness, which is stressful not only for patients, but also for family members. Numerous studies have demonstrated that family caregivers of persons with a severe mental illness suffer from significant stresses, experience moderately high levels of burden, and often receive inadequate assistance from mental health professionals. Effective family functioning in families with schizophrenia may be influenced by a variety of psychosocial factors. The purpose of this article was to present a review of the social science literature related to families living with schizophrenia that has been published during the last three decades. There is general agreement in the literature that a multitude of variables affect families with a severe mental illness, such as schizophrenia. Therefore, this literature review examined the most frequently investigated variables (coping, psychological distress and caregiver burden, social support, caregiver resiliency and depression, and client behavioral problems) as they are related to families and schizophrenia. PMID- 12554428 TI - Assessing patients' perception of self-care agency in psychiatric care. AB - Self-care agency is necessary for psychiatric patients to maintain a level of health that will enable them to have a satisfying personal life as well as to participate in their communities. This article discusses some theoretical and operational dimensions of self-care agency in patients treated for psychiatric illness, with an emphasis on self-care agency in mood disorder. Tools for measuring self-care agency based on Orem's power components are available, but tools specific to psychiatric patients' strengths and deficits, such as in the area of medication adherence and symptom management, need to be developed. PMID- 12554429 TI - Re-establishing psychodynamic theory as foundational knowledge for psychiatric/mental health nursing. AB - Psychiatric nursing care is based on an interpersonal process that relies on the therapeutic use of self in the service of patient care. Despite acknowledgment of the importance of the relationship, there has been little attention paid to the knowledge embedded in psychodynamic theory that can provide a basis for understanding the nurse-patient relationship. This paper argues that nurses need to understand and apply such theory if they are to pursue the assertion that the relationship with the patient is the foundation of their practice. The authors discuss the essential components of psychodynamic theory and their application to nursing as well as the possible reasons for the loss of interest in such theory. PMID- 12554432 TI - Basis for a proposed reference dose (RfD) for dioxin of 1-10 pg/kg-day: a weight of evidence evaluation of the human and animal studies. AB - The dioxins have been perhaps the most studied of all chemicals to which humans are routinely exposed. It has been reported that more than 5,000 scientific papers have been published that have evaluated the toxicology of 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD). Although the cancer hazard posed by this chemical has probably received the bulk of attention over the past 20 years, the U.S. Environmental Protection Agency (EPA) and the recent U.S. EPA Science Advisory Board (SAB) that reviewed the "Reassessment" have suggested that the noncancer hazard may well be more important than the cancer hazard at current background doses to the general public. The World Health Organization (WHO) and U.K. Food Standards Agency (FAO) committee (JECFA) on dioxins has reached similar conclusions. This article reviews the published studies involving laboratory animals and humans that address the noncancer effects. Based on our review, developmental toxicity is the most sensitive effect of TCDD consistently seen in mice and rats. Specifically, of the various studies, a no-observed-adverse effects level (NOAEL) of 13 ng/kg (maternal body burden) was identified as the most pertinent for deriving a reference dose (RfD) for humans. Although more than a dozen different adverse effects have been reported in various studies of humans over the past 25 years, the most consistent clinically important adverse effect of human exposure appears to be chloracne. Following a review of all published studies, we concluded that the best estimate of a LOAEL for production of chloracne is approximately 160 ng/kg (body burden). Based on our analysis, an RfD of between 1 and 10 pg/kg-d (TCDD TEQ) is consistent with the objectives of this risk criterion. Maintaining a lifetime average daily dose below this concentration, based on what is known today, should prevent noncancer effects in virtually all persons. This value is consistent with the JECFA recommendation of 70 pg/kg-mo. PMID- 12554433 TI - Genetic toxicity of naphthalene: a review. AB - Results of five previously unpublished studies of the genotoxicity of naphthalene are presented and extensively discussed in relation to the large database that exists in the published literature. According to the published literature, naphthalene has not induced gene mutations in bacterial assays or in a metabolically competent human cell line. However, naphthalene has caused cytotoxicity in some cell lines, and induced clastogenicity in Chinese hamster ovary (CHO) cells, in a human lymphoblastoid cell line, and in preimplantation mouse embryos. Some naphthalene metabolites were cytotoxic, but only naphthoquinones produced chromosomal damage in vitro. No chromosomal damage was observed in vivo in bone marrow erythrocytes from treated mice; however, a positive response was reported in a Drosophila assay for wing somatic mutation and recombination. The five unpublished studies of naphthalene genotoxicity include three studies in vitro (two Ames bacterial assays and an in vitro unscheduled DNA synthesis assay) and two in vivo (mouse micronucleus and in vivo unscheduled DNA synthesis). Naphthalene was inactive in all five studies, in agreement with reports in the published literature. Chronic inhalation of naphthalene over 2 yr induced an increased incidence of benign alveolar/bronchial adenomas in female mice, and nasal epithelial tumors in both sexes of rats. Inflammation, tissue damage, and subsequent regenerative hyperplasia at target organ sites occurred in both species. Results of standard genetic toxicity assays suggest that naphthalene is not likely to be genotoxic in vivo. Since the in vitro results come primarily from assays utilizing liver-mediated activation systems, and the in vivo results come from rodent organs that are not targets for tumors, tests using naphthalene-sensitive rodent tissues would determine the applicability of current data in addressing the mechanisms of these species and site-specific cancers. The standard assays reported here may be useful in predicting potential health hazard in other species, or in humans, in whom there are few reported instances of naphthalene-induced cancer, especially as more data on species-specific differences in naphthalene metabolism become available. Despite present data limitations, a threshold mechanism for tumorigenesis can be proposed. The absence of naphthalene-induced gene mutation and the presence of cytotoxicity and some chromosomal events in vitro are consistent with a threshold related mechanism of tumor induction, driven by cytotoxicity and cell regeneration, followed by genetic events, or by accumulation of naphthalene at specific target sites to allow in situ formation of a genotoxic metabolite to trigger or enhance spontaneous tumor development. PMID- 12554435 TI - Molecular phylogenetics of allodapine bees, with implications for the evolution of sociality and progressive rearing. AB - Allodapine bees have long been regarded as providing useful material for examining the origins of social behavior. Previous researchers have assumed that sociality arose within the Allodapini and have linked the evolution of sociality to a transition from mass provisioning to progressive provisioning of brood. Early phylogenetic studies of allodapines were based on morphological and life history data, but critical aspects of these studies relied on small character sets, where the polarity and coding of characters is problematic. We used nucleotide sequence data from one nuclear and two mitochondrial gene fragments to examine phylogenetic structure among nine allodapine genera. Our data set comprised 1506 nucleotide positions, of which 402 were parsimony informative. Maximum parsimony, log determinant, and maximum likelihood analyses produced highly similar phylogenetic topologies, and all analyses indicated that the tropical African genus Macrogalea was the sister group to all other allodapines. This finding conflicts with that of previous studies, in which Compsomelissa + Halterapis formed the most basal group. Changing the basal node of the Allodapini has major consequences for understanding evolution in this tribe. Our results cast doubt on the previous hypotheses that progressive provisioning and castelike social behavior evolved among lineages leading to the extant allodapine taxa. Instead, our results suggest that mass provisioning in Halterapis is a derived feature and that social behavior is an ancestral trait for all allodapine lineages. The forms of social behavior present in extant allodapines are likely to have resulted from a long evolutionary history, which may help explain the complexity of social traits found in many allodapine bees. PMID- 12554434 TI - Methyl parathion: a review of health effects. AB - Methyl parathion is an organophosphorus (OP) insecticide with insecticidal properties derived from acetylcholinesterase (AChE) inhibition; this same property is also the root of its toxicity in humans. Poisoning with methyl parathion leads to cholinergic overstimulation with signs of toxicity including sweating, dizziness, vomiting, diarrhea, convulsions, cardiac arrest, respiratory arrest, and, in extreme cases, death. Reports of methyl parathion intoxication, usually seen only in field pesticide applicators, have increased throughout the United States as a result of unauthorized application of methyl parathion inside homes. The health concerns of the use of methyl parathion have resulted in cancellation of its use in most food crops in the United States. This review examines the well-documented neurotoxicity of methyl parathion as well as effects on other organ systems. PMID- 12554436 TI - Partitioned likelihood support and the evaluation of data set conflict. AB - In simultaneous analyses of multiple data partitions, the trees relevant when measuring support for a clade are the optimal tree, and the best tree lacking the clade (i.e., the most reasonable alternative). The parsimony-based method of partitioned branch support (PBS) "forces" each data set to arbitrate between the two relevant trees. This value is the amount each data set contributes to clade support in the combined analysis, and can be very different to support apparent in separate analyses. The approach used in PBS can also be employed in likelihood: a simultaneous analysis of all data retrieves the maximum likelihood tree, and the best tree without the clade of interest is also found. Each data set is fitted to the two trees and the log-likelihood difference calculated, giving "partitioned likelihood support" (PLS) for each data set. These calculations can be performed regardless of the complexity of the ML model adopted. The significance of PLS can be evaluated using a variety of resampling methods, such as the Kishino-Hasegawa test, the Shimodiara-Hasegawa test, or likelihood weights, although the appropriateness and assumptions of these tests remains debated. PMID- 12554437 TI - Phylogeny of eusocial Lasioglossum reveals multiple losses of eusociality within a primitively eusocial clade of bees (Hymenoptera: Halictidae). AB - We performed a phylogenetic analysis of the species, species groups, and subgenera within the predominantly eusocial lineage of Lasioglossum (the Hemihalictus series) based on three protein coding genes: mitochondrial cytochrome oxidase I, nuclear elongation factor 1alpha and long-wavelength rhodopsin. The entire data set consisted of 3421 aligned nucleotide sites, 854 of which were parsimony informative. Analyses by equal weights parsimony, maximum likelihood, and Bayesian methods yielded good resolution among the 53 taxa/populations, with strong bootstrap support and high posterior probabilities for most nodes. There was no significant incongruence among genes, and parsimony, maximum likelihood, and Bayesian methods yielded congruent results. We mapped social behavior onto the resulting tree for 42 of the taxa/populations to infer the likely history of social evolution within Lasioglossum. Our results indicate that eusociality had a single origin within Lasioglossum. Within the predominantly eusocial clade, however, there have been multiple (six) reversals from eusociality to solitary nesting, social polymorphism, or social parasitism, suggesting that these reversals may be more common in primitively eusocial Hymenoptera than previously anticipated. Our results support the view that eusociality is hard to evolve but easily lost. This conclusion is potentially important for understanding the early evolution of the advanced eusocial insects, such as ants, termites, and corbiculate bees. PMID- 12554438 TI - When do parasites fail to speciate in response to host speciation? AB - Cospeciation generally increases the similarity between host and parasite phylogenies. Incongruence between host and parasite phylogenies has previously been explained in terms of host switching, sorting, and duplication events. Here, we describe an additional process, failure of the parasite to speciate in response to host speciation, that may be important in some host-parasite systems. Failure to speciate is likely to occur when gene flow among parasite populations is much higher than that of their hosts. We reconstructed trees from mitochondrial and nuclear DNA sequences for pigeons and doves (Aves: Columbiformes) and their feather lice in the genus Columbicola (Insecta: Phthiraptera). Although comparisons of the trees from each group revealed a significant amount of cospeciation, there was also a significant degree of incongruence. Cophylogenetic analyses generally indicated that host switching may be an important process in the history of this host-parasite association. Using terminal sister taxon comparisons, we also identified three apparent cases where the host has speciated but the associated parasite has not. In two of these cases of failure to speciate, these comparisons involve allopatric sister taxa of hosts whose lice also occur on hosts sympatric with both of the allopatric sisters. These additional hosts for generalist lice may promote gene flow with lice on the allopatric sister species. Relative rate comparisons for the mitochondrial cytochrome oxidase I gene indicate that molecular substitution occurs about 11 times faster in lice than in their avian hosts. PMID- 12554439 TI - Testing a molecular clock without an outgroup: derivations of induced priors on branch-length restrictions in a Bayesian framework. AB - We propose a Bayesian method for testing molecular clock hypotheses for use with aligned sequence data from multiple taxa. Our method utilizes a nonreversible nucleotide substitution model to avoid the necessity of specifying either a known tree relating the taxa or an outgroup for rooting the tree. We employ reversible jump Markov chain Monte Carlo to sample from the posterior distribution of the phylogenetic model parameters and conduct hypothesis testing using Bayes factors, the ratio of the posterior to prior odds of competing models. Here, the Bayes factors reflect the relative support of the sequence data for equal rates of evolutionary change between taxa versus unequal rates, averaged over all possible phylogenetic parameters, including the tree and root position. As the molecular clock model is a restriction of the more general unequal rates model, we use the Savage-Dickey ratio to estimate the Bayes factors. The Savage-Dickey ratio provides a convenient approach to calculating Bayes factors in favor of sharp hypotheses. Critical to calculating the Savage-Dickey ratio is a determination of the prior induced on the modeling restrictions. We demonstrate our method on a well-studied mtDNA sequence data set consisting of nine primates. We find strong support against a global molecular clock, but do find support for a local clock among the anthropoids. We provide mathematical derivations of the induced priors on branch length restrictions assuming equally likely trees. These derivations also have more general applicability to the examination of prior assumptions in Bayesian phylogenetics. PMID- 12554440 TI - Assessing progress in systematics with continuous jackknife function analysis. AB - Systematists expect their hypotheses to be asymptotically precise. As the number of phylogenetically informative characters for a set of taxa increases, the relationships implied should stabilize on some topology. If true, this increasing stability should clearly manifest itself if an index of congruence is plotted against the accumulating number of characters. Continuous jackknife function (CJF) analysis is a new graphical method that portrays the extent to which available data converge on a specified phylogenetic hypothesis, the reference tree. The method removes characters with increasing probability, analyzes the rarefied data matrices phylogenetically, and scores the clades shared between each of the resulting trees and the reference tree. As more characters are removed, the number of shared clades must decrease, but the rate of decrease will depend on how decisively the data support the reference tree. Curves for stable phylogenies are clearly asymptotic with nearly 100% congruence for a substantial part of the curve. Less stable phylogenies lose congruent nodes quickly as characters are excluded, resulting in a more linear or even a sigmoidal relationship. Curves can be interpreted as predictors of whether the addition of new data of the same type is likely to alter the hypothesis under test. Continuous jackknife function analysis makes statistical assumptions about the collection of character data. To the extent that CJF curves are sensitive to violations of unbiased character collection, they will be misleading as predictors. Convergence of data on a reference tree does not guarantee historical accuracy, but it does predict that the accumulation of further data under the sampling model will not lead to rapid changes in the hypothesis. PMID- 12554441 TI - Computing the uniform component of shape variation. AB - Any change in shape of a configuration of landmark points in two or three dimensions includes a uniform component, a component that is a wholly linear (affine) transformation. The formulas for estimating this component have been standardized for two-dimensional data but not for three-dimensional data. We suggest estimating the component by way of the complementarity between the uniform component and the space of partial warps. The component can be estimated by regression in either one space or the other: regression on the partial warps, followed by their removal, or regression on a basis for the uniform component itself. Either of the new methods can be used for both two- and three-dimensional landmark data and thus generalize Bookstein's (1996, pages 153-168 in Advances in morphometrics [L. F. Marcus et al., eds.], Plenum, New York) linearized Procrustes formula for estimating the uniform component in two dimensions. PMID- 12554442 TI - Speciation on a conveyor belt: sequential colonization of the hawaiian islands by Orsonwelles spiders (Araneae, Linyphiidae). AB - Spiders of the recently described linyphiid genus Orsonwelles (Araneae, Linyphiidae) are one of the most conspicuous groups of terrestrial arthropods of Hawaiian native forests. There are 13 known Orsonwelles species, and all are single- island endemics. This radiation provides an excellent example of insular gigantism. We reconstructed the cladistic relationships of Orsonwelles species using a combination of morphological and molecular characters (both mitochondrial and nuclear sequences) within a parsimony framework. We explored and quantified the contribution of different character partitions and their sensitivity to changes in the traditional parameters (gap, transition, and transversion costs). The character data show a strong phylogenetic signal, robust to parameter changes. The monophyly of the genus Orsonwelles is strongly supported. The parsimony analysis of all character evidence combined recovered a clade with of all the non-Kauai Orsonwelles species; the species from Kauai form a paraphyletic assemblage with respect to the latter former clade. The biogeographic pattern of the Hawaiian Orsonwelles species is consistent with colonization by island progression, but alternative explanations for our data exist. Although the geographic origin of the radiation remains unknown, it appears that the ancestral colonizing species arrived first on Kauai (or an older island). The ambiguity in the area cladogram (i.e., post-Oahu colonization) is not derived from conflicting or unresolved phylogenetic signal among Orsonwelles species but rather from the number of taxa on the youngest islands. Speciation in Orsonwelles occurred more often within islands (8 of the 12 cladogenic events) than between islands. A molecular clock was rejected for the sequence data. Divergence times were estimated by using the nonparametric rate smoothing method of Sanderson (1997, Mol. Biol. Evol. 14:1218-1231) and the available geological data for calibration. The results suggest that the oldest divergences of Orsonwelles spiders (on Kauai) go back about 4 million years. PMID- 12554443 TI - Molecular systematics and adaptive radiation of Hawaii's endemic Damselfly genus Megalagrion (Odonata: Coenagrionidae). AB - Damselflies of the endemic Hawaiian genus Megalagrion have radiated into a wide variety of habitats and are an excellent model group for the study of adaptive radiation. Past phylogenetic analysis based on morphological characters has been problematic. Here, we examine relationships among 56 individuals from 20 of the 23 described species using maximum likelihood (ML) and Bayesian phylogenetic analysis of mitochondrial (1287 bp) and nuclear (1039 bp) DNA sequence data. Models of evolution were chosen using the Akaike information criterion. Problems with distant outgroups were accommodated by constraining the best ML ingroup topology but allowing the outgroups to attach to any ingroup branch in a bootstrap analysis. No strong contradictions were obtained between either data partition and the combined data set. Areas of disagreement are mainly confined to clades that are strongly supported by the mitochondrial DNA and weakly supported by the elongation factor 1alpha data because of lack of changes. However, the combined analysis resulted in a unique tree. Correlation between Bayesian posterior probabilities and bootstrap percentages decreased in concert with decreasing information in the data partitions. In cases where nodes were supported by single characters bootstrap proportions were dramatically reduced compared with posterior probabilities. Two speciation patterns were evident from the phylogenetic analysis. First, most speciation is interisland and occurred as members of established ecological guilds colonized new volcanoes after they emerged from the sea. Second, there are several instances of rapid radiation into a variety of specialized habitats, in one case entirely within the island of Kauai. Application of a local clock procedure to the mitochondrial DNA topology suggests that two of these radiations correspond to the development of habitat on the islands of Kauai and Oahu. About 4.0 million years ago, species simultaneously moved into fast streams and plant leaf axils on Kauai, and about 1.5 million years later another group moved simultaneously to seeps and terrestrial habitats on Oahu. Results from the local clock analysis also strongly suggest that Megalagrion arrived in Hawaii about 10 million years ago, well before the emergence of Kauai. Date estimates were more sensitive to the particular node that was fixed in time than to the model of local branch evolution used. We propose a general model for the development of endemic damselfly species on Hawaiian Islands and document five potential cases of hybridization (M. xanthomelas x M. pacificum, M. eudytum x M. vagabundum, M. orobates x M. oresitrophum, M. nesiotes x M. oahuense, and M. mauka x M. paludicola). PMID- 12554444 TI - The combinatorics of tandem duplication trees. AB - We developed a recurrence relation that counts the number of tandem duplication trees (either rooted or unrooted) that are consistent with a set of n tandemly repeated sequences generated under the standard unequal recombination (or crossover) model of tandem duplications. The number of rooted duplication trees is exactly twice the number of unrooted trees, which means that on average only two positions for a root on a duplication tree are possible. Using the recurrence, we tabulated these numbers for small values of n. We also developed an asymptotic formula that for large n provides estimates for these numbers. These numbers give a priori probabilities for phylogenies of the repeated sequences to be duplication trees. This work extends earlier studies where exhaustive counts of the numbers for small n were obtained. One application showed the significance of finding that most maximum-parsimony trees constructed from repeat sequences from human immunoglobins and T-cell receptors were tandem duplication trees. Those findings provided strong support to the proposed mechanisms of tandem gene duplication. The recurrence relation also suggests efficient algorithms to recognize duplication trees and to generate random duplication trees for simulation. We present a linear-time recognition algorithm. PMID- 12554445 TI - Taxon sampling, bioinformatics, and phylogenomics. PMID- 12554446 TI - Is sparse taxon sampling a problem for phylogenetic inference? PMID- 12554447 TI - Branch lengths and support. PMID- 12554448 TI - Using tree shape. PMID- 12554449 TI - Is cladogenesis heritable? AB - The heritability of speciation rates and extinction risks is a crucial parameter in models of macroevolution, but little direct evidence has been available to assess the occurrence, strength, or generality of this heritability. We tested for heritability using correlations between ancestral and descendent branch lengths in phylogenetic trees, an approach first applied to a bird phylogeny by Harvey et al. (1991, pages 123-137 in Genes in ecology [R. J. Berry et al., eds.], Blackwell Scientific, Oxford). We applied Harvey et al.'s test to some of the largest DNA sequence-based phylogenetic analyses published to date for plants, insects, fungi, and bacteria. If one of two parent lineages splits first and if this is the case for any heritable reason, then on average we expect its daughter lineages to also split first. We also used a randomization procedure to assess significance of branch length heritability. Using maximum parsimony and maximum likelihood branch lengths and trees made ultrametric after nonparametric rate smoothing or by enforcing a molecular clock, we found a pattern for most clades consistent with heritable net cladogenesis. Heritability of cladogenesis may be a general phenomenon, detectable across a large number of lineages and a broad range of taxa. PMID- 12554450 TI - Phylogeny imbalance: taxonomic level matters. AB - Two lines of evidence indicate that the degree of symmetry in phylogenetic topologies differs at different hierarchical levels. First, in a set of 61 phylogenies with superspecific taxa as their terminals, trees were on average more unbalanced (asymmetric) when the species richness of terminals was considered than when it was not. Second, nodes with a given number of higher taxa descended from them were on average more unbalanced than were nodes with the same number of species as descendants. There are several possible reasons--some biological, some artifactual--for the differences. Whatever the reason, these results caution against treating species-level and higher level phylogenies as equivalent when considering tree shape. The imbalance measure adopted here permits the use of trees that contain polytomies, facilitating a larger sample than has been achieved previously. PMID- 12554451 TI - Whole-tree methods for detecting differential diversification rates. AB - Prolific cladogenesis, adaptive radiation, species selection, key innovations, and mass extinctions are a few examples of biological phenomena that lead to differential diversification among lineages. Central to the study of differential diversification rates is the ability to distinguish chance variation from that which requires deterministic explanation. To detect diversification rate variation among lineages, we propose a number of methods that incorporate information on the topological distribution of species diversity from all internal nodes of a phylogenetic tree. These whole-tree methods (M(Pi), M(Sigma), and M(R)) are explicitly connected to a null model of random diversification--the equal-rates Markov (ERM) random branching model--and an alternative model of differential diversification: M(Pi) is based on the product of individual nodal ERM probabilities; M(Sigma) is based on the sum of individual nodal ERM probabilities, and M(R) is based on a transformation of ERM probabilities that corresponds to a formalized system that orders trees by their relative symmetry. These methods have been implemented in a freely available computer program, SYMMETREE, to detect clades with variable diversification rates, thereby allowing the study of biological processes correlated with and possibly causal to shifts in diversification rate. Application of these methods to several published phylogenies demonstrates their ability to contend with relatively large, incompletely resolved trees. These topology-based methods do not require estimates of relative branch lengths, which should facilitate the analysis of phylogenies, such as supertrees, for which such data are unreliable or unavailable. PMID- 12554452 TI - Power of eight tree shape statistics to detect nonrandom diversification: a comparison by simulation of two models of cladogenesis. AB - We used simulations to compare the relative power of eight statistical tests to detect imbalance in phylogenies that is too great to be ascribed to an equal rates Markov null model. Three of these tests have never had their power assessed before. Our simulations are the first to assess performance under scenarios in which the speciation rates of various lineages can evolve independently. In one of the scenarios explored, rates depend upon the value of an evolving trait, whereas in the other the probability that a species will speciate declines with the time since it last did so. The results indicate that the relative performance of the methods depends upon how the imbalance is generated. Different types of processes lead to different imbalance signatures, i.e., different patterns of imbalance at different depths in the phylogeny, and the measures of tree shape differ in the depth of phylogeny at which they are most sensitive. Relative performance is also affected by tree size but does not appear to depend greatly upon the degree of speciation rate variation among lineages. Two of the indices (Colless's index I(c) and Shao and Sokal's Nmacr;) show reasonable performance throughout, but another (Shao and Sokal's B(2)) is never indicated to be a preferred method. Two tests that do not require completely resolved phylogenies, mean I' and mean I'(10), have reasonable power. PMID- 12554453 TI - Phylogeny shape and the phylogenetic comparative method. AB - We explored the impact of phylogeny shape on the results of interspecific statistical analyses incorporating phylogenetic information. In most phylogenetic comparative methods (PCMs), the phylogeny can be represented as a relationship matrix, and the hierarchical nature of interspecific phylogenies translates into a distinctive blocklike matrix that can be described by its eigenvectors (topology) and eigenvalues (branch lengths). Thus, differences in the eigenvectors and eigenvalues of different relationship matrices can be used to gauge the impact of possible phylogeny errors by comparing the actual phylogeny used in a PCM analysis with a second phylogenetic hypothesis that may be more accurate. For example, we can use the sum of inverse eigenvalues as a rough index to compare the impact of phylogenies with different branch lengths. Topological differences are better described by the eigenvectors. In general, phylogeny errors that involve deep splits in the phylogeny (e.g., moving a taxon across the base of the phylogeny) are likely to have much greater impact than will those involving small perturbations in the fine structure near the tips. Small perturbations, however, may have more of an impact if the phylogeny structure is highly dependent (with many recent splits near the tips of the tree). Unfortunately, the impact of any phylogeny difference on the results of a PCM depends on the details of the data being considered. Recommendations regarding the choice, design, and statistical power of interspecific analyses are also made. PMID- 12554454 TI - New inferences from tree shape: numbers of missing taxa and population growth rates. AB - The relative positions of branching events in a phylogeny contain information about evolutionary and population dynamic processes. We provide new summary statistics of branching event times and describe how these statistics can be used to infer rates of species diversification from interspecies trees or rates of population growth from intraspecies trees. We also introduce a phylogenetic method for estimating the level of taxon sampling in a clade. Different evolutionary models and different sampling regimes can produce similar patterns of branching events, so it is important to consider explicitly the model assumptions involved when making evolutionary inferences. Results of an analysis of the phylogeny of the mosquito-borne flaviviruses suggest that there could be several thousand currently unidentified viruses in this clade. PMID- 12554455 TI - Signatures of random and selective mass extinctions in phylogenetic tree balance. AB - Current models of diversification with evolving speciation rates have trouble mimicking the extreme imbalance seen in estimated phylogenies. However, these models have not incorporated extinction. Here, we report on a simple simulation model that includes heritable and evolving speciation rates coupled with mass extinctions, Random (but not selective) mass extinctions, coupled with evolving among-lineage variation in speciation rates, increase imbalance of postrecovery clades. Thus, random mass extinctions are plausible contributors to the imbalance of modern clades. Paleontological evidence suggests that mass extinctions are often random with respect to ecological and morphological traits, consistent with our simulations. In contrast, evidence that the current anthropogenic mass extinction is phylogenetically selective suggests that the current extinction episode may be qualitatively different from past ones in the way it reshapes future biotas. PMID- 12554456 TI - Phylogenetic balance and ecological evenness. AB - The frequency distribution of numbers of species in taxonomic groups, where many species belong to a few very diverse higher taxa, is mirrored by that of species in most communities, where many individuals belong to a few very abundant species. Various hypotheses mechanistically link a species' community abundance with the diversity of the higher level taxon (genus, family, order) to which it belongs, but empirical data are equivocal about general trends in the relation between rank-taxon diversity and mean abundance. One reason for this inconclusive result may be the effect of the semisubjective nature of rank-based classification. We assessed the relationship between clade diversity and mean species abundance for two diverse tropical tree communities, using both traditional rank-based analysis and two new phylogenetic analyses (based on the ratio of individuals to taxa at each node in the phylogeny). Both rank-based and phylogenetic analyses using taxonomic ranks above the species level as terminal taxa detected a trend associating common species with species-rich families. In contrast, phylogenetic analyses using species as terminal taxa could not distinguish the observed distribution of species abundances from a random distribution with respect to clade diversity. The difference between these results might be due to (1) the absence of a real phylogeny-wide relationship between clade abundance and diversity, (2) the influence of poor phylogenetic resolution within families in our phylogenies, or (3) insufficient sensitivity of our metrics to subtle tree-wide effects. Further development and application of phylogeny-based methods for testing abundance-diversity relationships is needed. PMID- 12554457 TI - Tree balance, time slices, and evolutionary turnover in cretaceous planktonic foraminifera. AB - Studies of phylogenetic tree shape often concentrate on the balance of phylogenies of extant taxa. Paleontological phylogenies (which include extinct taxa) can contain additional useful information and can directly document changes in tree shape through evolutionary time. Unfortunately, the inclusion of extinct taxa lowers the power of direct examinations of tree balance because it increases the range of tree shapes expected under null models of evolution (with equal rates of speciation and extinction across lineages). A promising approach for the analysis of tree shape in paleontological phylogenies is to break the phylogeny down into time slices, examining the shape of the phylogeny of taxa alive at each time slice and changes in that shape between successive time slices. This method was illustrated with 57 time slices through a stratophenetic phylogeny of the Cretaceous planktonic foraminiferal superfamily Globotruncanacea. At 3 of 56 intervals between time slices, 93-92.5 million years ago (MYA), 89-88.5 MYA, and 85.5-84 MYA, the group showed steep increases in imbalance. Although none of these increases were significant after Bonferroni correction, these points in the history of the Globotruncanacea were nevertheless identified as deserving of further macroevolutionary investigation. The 84 MYA time slice coincides with a peak in species turnover for the superfamily. Time slices through phylogenies may prove useful for identifying periods of time when evolution was proceeding in a nonstochastic manner. PMID- 12554458 TI - Contingent predictability in evolution: key traits and diversification. AB - Key innovations have often been invoked to explain the exceptional diversification of particular groups. However, there are few convincing examples of traits that are repeatedly and consistently associated with increased diversification. The paucity of such cases may reflect the contingent nature of the diversifying effect of key traits. These contingencies can be viewed as statistical interactions between the trait and at least three kinds of factors: (1) other taxa, (2) other traits of the group itself, and (3) the physical environment. I describe tentative examples in each of these categories: (1) a dampening of the diversification of clades with image-forming eyes by groups that earlier evolved such eyes, (2) an effect of growth form (woody or herbaceous) on the diversifying effect of biotic seed dispersal in angiosperms, and (3) an effect of atmospheric CO(2) level on the diversifying effect of C(4) photosynthesis in monocots. These examples suggest the need for more complex analyses of the relationship between possible key traits and diversification. They also suggest that radiations may be predictable given certain circumstances, thus supporting a view of evolution as both predictable and contingent. Ironically, a certain degree of predictability may be critical to arguments for evolutionary contingency. PMID- 12554459 TI - Molecular phylogeny of grouse: individual and combined performance of W-linked, autosomal, and mitochondrial loci. AB - The phylogeny of grouse (Aves: Tetraoninae) was reconstructed using four noncoding loci: two were W-linked, one was autosomal, and one was the mitochondrial control region (CR). The rapidly evolving CR provided resolution throughout the tree, whereas the slowly evolving nuclear loci failed to resolve deeper nodes. The tree based on all four loci combined was almost identical to the CR tree and did not improve resolution or bootstrap support. The stemminess and imbalance of the trees were good determinants of the quality of the phylogenetic signal. The skewness of the tree score distribution (g(1)) behaved contrary to prediction; loci that had a more symmetric tree score distribution produced trees that had greater stemminess and balance. The quality of the phylogenetic signal was related to the evolutionary rate. Four clades of grouse were discovered. Two of these clades corresponded to currently recognized genera Bonasa and Lagopus. Bonasa was the sister to other grouse and Lagopus was the sister to the other two non-Bonasa clades. The third clade included Falcipennis, Tetrao, and Lyrurus. The fourth clade included the genera Centrocercus, Dendragapus, and Tympanuchus. The data support recognition of Falcipennis canadensis franklinii and Dendragapus obscurus fuliginosus as species. PMID- 12554460 TI - Combined nuclear and mitochondrial DNA sequences resolve generic relationships within the Cracidae (Galliformes, Aves). AB - The Cracidae is one of the most endangered and distinctive bird families in the Neotropics, yet the higher relationships among taxa remain uncertain. The molecular phylogeny of its 11 genera was inferred using 10,678 analyzable sites (5,412 from seven different mitochondrial segments and 5,266 sites from four nuclear genes). We performed combinability tests to check conflicts in phylogenetic signals of separate genes and genomes. Phylogenetic analysis showed that the unrooted tree of ((curassows, horned guan) (guans, chachalacas)) was favored by most data partitions and that different data partitions provided support for different parts of the tree. In particular, the concatenated mitochondrial DNA (mtDNA) genes resolved shallower nodes, whereas the combined nuclear sequences resolved the basal connections among the major clades of curassows, horned guan, chachalacas, and guans. Therefore, we decided that for the Cracidae all data should be combined for phylogenetic analysis. Maximum parsimony (MP), maximum likelihood (ML), and Bayesian analyses of this large data set produced similar trees. The MP tree indicated that guans are the sister group to (horned guan, (curassows, chachalacas)), whereas the ML and Bayesian analysis recovered a tree where the horned guan is a sister clade to curassows, and these two clades had the chachalacas as a sister group. Parametric bootstrapping showed that alternative trees previously proposed for the cracid genera are significantly less likely than our estimate of their relationships. A likelihood ratio test of the hypothesis of a molecular clock for cracid mtDNA sequences using the optimal ML topology did not reject rate constancy of substitutions through time. We estimated cracids to have originated between 64 and 90 million years ago (MYA), with a mean estimate of 76 MYA. Diversification of the genera occurred approximately 41-3 MYA, corresponding with periods of global climate change and other Earth history events that likely promoted divergences of higher level taxa. PMID- 12554461 TI - A phylogeny of the tinamous (aves: palaeognathiformes) based on integumentary characters. AB - A cladistic analysis of the tinamous, including the 47 currently recognized species and some distinct subspecies, was conducted based on 80 integumentary characters from adult and natal plumage, ramphoteca (corneum sheath of bill), and podoteca (horny scales of legs). For the adult plumage (50 characters), we studied feather pigmentation patterns from different pterylae (feather tracts). A criterion of overlap of basic pigmentation elements was used to assign costs to the transformation between the states in most of these characters in such a way that transformations between more similar conditions were less costly. The consensus tree was almost fully resolved, and about 50% of its groups were relatively well supported. Because the only outgroup that could be used provided a poor root, two possible rootings of the ingroup subtree were considered; in both cases, only one of the two traditional subfamilies (the steppe tinamous) was recovered, and the other (the forest tinamous) appeared as paraphyletic. The results of the present analysis are compared with those from an osteological data set, using a strict supertree technique. The combined tree has a large number of nodes, indicating a high degree of congruence between the two data sets. PMID- 12554518 TI - Odile Schweisguth: pioneer of European pediatric oncology. PMID- 12554519 TI - A novel idea for an organized hospital/school program for children with malignancies: issues in implementation. AB - The aim of this pilot study was to provide children with cancer hospitalized at the Department of Pediatric Hematology-Oncology of the University Hospital of Heraklion, Crete, with an educational program based on the SIOP guidelines. This would help them to follow the school program during the days of absence due to treatment. The realization of the initial idea generated difficulties that had to be overcome to achieve the goals and caused several changes in the program. Children refused to attend a regular class in the appropriately designed playroom of the Department but were very keen on room-to-room teaching that was adjusted to their needs and wills. This method of teaching was very well accepted by the children. A workup was done with parents and health professionals to convince them of the necessity of such a program and persuade them to participate in the procedure by encouraging children to attend it. Several steps have been taken and others have been planned for the teachers to receive education concerning the special needs of these children after dismissal from the hospital as well as that of their siblings to help in the reentry procedure. An adjustment to the personal needs of the hospitalized children with cancer, especially in areas where the psychosocial resources are minor, has proved very helpful and effective for the reentry of these children to regular school and social life. PMID- 12554520 TI - Long-term sequelae in children treated for brain tumors: impairments, disability, and handicap. AB - Sixty-one long-term survivors, treated for brain tumors in childhood, were evaluated in term of neurological impairments, disability, and handicap. Thirty eight patients (pts) (62%) had at least one impairment. Visual impairment was detected in 14 pts (24%), associated with recurrence (p = .012). Thirty-four patients (56%) had motor impairment, associated with sex (female) in irradiated patients; 13 (21%) had epilepsy, associated with supratentorial tumor site (p = .001). The same number of patients had brain atrophy; risk factors were hydrocephalus at diagnosis and perioperative complications. Sixteen patients (30%) had IQ score < 80, associated with young age at first treatment (p = .006) and recurrence (p = .043). Twenty-seven out of 61 of our patients (44%) were disabled: 12 mildly, 14 moderately, and 1 severely. Epilepsy was the most important risk factor for disability. Cognitive impairment, motor impairment, and epilepsy were associated with employment (43%); cognitive impairment was also associated with education. PMID- 12554521 TI - High survival rate in childhood non-Hodgkin lymphoma without CNS involvement: results of BFM 95 study in Kuwait. AB - Non-Hodgkin lymphomas (NHL) in children are the second most common malignant tumors in Kuwait. Until 1995 the patients (pts) received institutional protocols. From October 1995 to September 2000 21 children with NHL were treated. Five children were treated by NHL BFM 90 protocol, 7 pts received NHL BFM 95 scheme, and 9 children underwent therapy abroad or according to different types of protocols. The results of a retrospective analysis of NHL BFM 95 protocol in Kuwait are reported. Seven patients diagnosed with NHL--group B: 3 children with Burkitt lymphoma (B-cell NHL) and group A: 4 children with lymphoblastic lymphoma (T-cell NHL)--were treated from October 1995 to September 2000 in the Kuwait Cancer Control Centre according to NHL BFM 95 protocol. Group B consisted of 2 girls and 1 boy; median age at diagnosis was 4 years 8 months, 2 pts classified as stage II and 1 pt as stage III. All patients were assigned to risk group R2. Median follow-up is 2 years 8 months. Group A included 1 girl and 3 boys; median age at diagnosis was 5 years 8 months, 1 pt classified as stage III and 3 pts as stage IV. All patients were assigned to IR group. Median follow-up is 3 years 6 months. In group B all 3 pts are in 1st CR; in group A 3 pts are in 1st CR and 1 pt having Li-Fraumani syndrome died after the 3rd relapse of disease during therapy. In both groups there was no toxic death, myelotoxicity WHO grade III-IV, hepatotoxicity WHO grade II-III. Treatment results of NHL BFM 95 study in our small group of patients are very optimistic. Six patients are in 1st CR and one died due to progression of disease. Despite the small group of patients, the results suggest that NHL BFM 95 protocol is highly effective and safe with regular supportive care. PMID- 12554522 TI - Palliative radiotherapy in children with neuroblastoma. AB - Limited information is available regarding the efficacy of external beam radiation therapy in the palliation of metastatic disease from neuroblastoma. From 1960 to 2000, 29 children with 53 metastatic sites received palliative radiotherapy. There were 26 soft tissue (group I), 19 bone (group II), 5 brain (group III), and 3 hepatic (group IV) treated sites. Median radiotherapy doses for groups I, II, III, and IV sites were 2000, 2000, 2400, and 450 cGy, respectively. For group I sites, complete response was complete disappearance of mass, partial response was > or =50% resolution of mass, no response was <50% resolution or < or =25% progression of mass, and progressive disease was >25% progression of mass. For group II sites, complete response was complete pain relief without medication, partial response was > or =50% pain relief with or without medication, no response was <50% change in pain with medication, and progressive disease was increase in pain and/or medication. Median survival was 2.5 months after palliative radiotherapy. For group I sites, complete response was seen in 1 (4%) while partial response was documented in 19 (73%). Duration of response was until death in 18 responders (90%); 2 patients relapsed with an increasing soft tissue mass at 5 months and 1 year after palliative radiotherapy. For group II sites, complete response was seen in 8 (42%) while partial response was documented in 7 (37%). Duration of response was until death in 14 responders (93%); 1 patient had relapse of pain 1 year after palliative radiotherapy. For the 5 group III children, the median survival was 2.5 months with a range of 2 days to 13 months. Four children had neurological improvement after cranial radiotherapy; one patient progressed and died secondary to neurological compromise 2 days after radiotherapy. For the 3 group IV sites, 2 had improvement in respiratory status after radiotherapy, whereas 1 progressed despite doxorubicin, cyclophosphamide, and radiation. The only patient who survived had a stage IV-S neuroblastoma with liver metastases and is alive 13 years after hepatic irradiation. Radiotherapy is an effective treatment for palliation of symptomatic metastatic disease in children with neuroblastoma. PMID- 12554523 TI - Stable transgenic expression of IL-2 and HSV1-tk by single and fusion tumor cell lines bearing EWS/FLI-1 chimeric genes. AB - In augmenting systemic anti-tumor immune response, the authors evaluated the genetic modification of Ewing family tumor (EFT) cell lines for use as allogeneic vaccines. EFT cell lines A673 and RD-ES were transfected with cDNAs for human interleukin (IL)-2 and/or HSV1 thymidine kinase (HSV1-tk), respectively. Clones with high and stable secretion of IL-2 alone or with coexpression of functional HSV1-tk were obtained and their features were analyzed. IL-2 expressing clones derived from the A673 cell line demonstrated decreased expression of HLA class I molecules compared with the parental cell line and corresponding clones derived from RD-ES. However, IFN-gamma could upregulate the expression of HLA class I antigens by IL-2 transfected A673 cells. Ganciclovir induced apoptosis in double transfected cell clones. IL-2/HSV1-tk cells continued to produce and release IL-2 after initial ganciclovir treatment. After gamma-irradiation, transfected clones released bioactive IL-2 in a quantity sufficient to activate T and natural killer cells in culture. A polyvalent allogeneic vaccine was also obtained using fusion of two different transgenic cell lines. The resulting hybrids inherited antigenic and transgenic characteristics of both parental cell lines. It is presumed that the cell lines generated here could be used as allogeneic vaccines for treatment of patients with EFTs. PMID- 12554524 TI - Malignant melanoma of the soft parts showing calciphylaxis. AB - Calciphylaxis is a poorly understood and highly morbid syndrome of vascular calcification and skin necrosis. Hypercalcemia, hyperphosphatemia, and secondary hyperparathyroidism are the factors implicated in the pathogenesis of calciphylaxis, which is generally identified in patients with hyperparathyroidism secondary to end-stage renal disease. It has also been observed in primary hyperparathyroidism, end-stage liver disease, and rheumatoid arthritis, in the absence of renal disease. There are few case reports of calciphylaxis occurring in hypercalcemia of malignancy. An unusual case is reported of calciphylaxis associated with malignant melanoma of the soft parts in the absence of renal or parathyroid disease. This is the first reported case of this soft tissue sarcoma showing calciphylactic changes. PMID- 12554525 TI - Acute perforative appendicitis during preoperative chemotherapy for Wilms tumor. AB - Infectious complications are not uncommon in children undergoing treatment for cancer. Abdominal pain, especially right lower quadrant pain secondary to appendiceal and cecal inflammation, is a major concern in immunocompromised hosts and a potential source of sepsis. The authors report the case of a child who developed acute perforative appendicitis requiring appendectomy while on preoperative chemotherapy for Wilms tumor, stage IV, favorable histology. Problems related to diagnosis and management of acute abdominal pain and infection in an immunocompromised child with an abdominal mass are discussed along with a review of the literature. PMID- 12554526 TI - Regression of refractory rhabdomyosarcoma after allogeneic stem-cell transplantation. AB - A 17-year-old girl developed refractory rhabdomyosarcoma. An allogeneic peripheral-blood stem-cell transplant was performed after a myeloablative regimen. Although rapid disease progression had resolved transiently, after the start of high-dose chemotherapy, re-progression was apparently observed from day 14. However, delayed tumor regression occurred on day 30, shortly after the reduction of immunosuppressants. She achieved a partial remission. The second tumor regression provides suggestive clinical evidence that graft-versus-tumor effect may occur against rhabdomyosarcoma. Although further investigation is required, allogeneic stem-cell transplantation could provide a new therapeutic option for refractory rhabdomyosarcoma. PMID- 12554527 TI - Benign intracranial hypertension in association with acute lymphoblastic leukemia. PMID- 12554528 TI - Recombinant interferon alfa 2a in hepatic hemangiomatosis with congestive heart failure: a case report. AB - A 45-day-old patient was admitted with dyspnea, hepatomegaly, tachycardia, holosystolic murmur in the precordial region, and continuous murmur at the right hypochondrium. Four cutaneous angiomas were noted. Instrumental examinations revealed congestive heart failure and multiple focal lesions in the liver with typical features of hemangiomas. The therapy with subcutaneous interferon-alfa-2a (IFN-alpha) was administered for 12 months with progressive regression of cutaneous hemangiomas, liver lesions, and cardiological alterations. IFN-alpha therapy was effective without any significant adverse effects. PMID- 12554529 TI - Evans syndrome in a patient with chromosome 22q11.2 deletion syndrome: a case report. AB - One patient with a chromosome 22q11.2 deletion and Evans syndrome is reported in this paper. Microdeletions of 22q11.2 are the main etiology for DiGeorge syndrome, a disorder characterized by heart defects, immune deficiencies due to aplasia or hypoplasia of the thymus, and hypocalcemia. Evans syndrome refers to a hematological autoimmune disorder with autoimmune hemolytic anemia accompanied by immune thrombocytopenia. A wide range of autoimmune disorders have been described in DiGeorge syndrome and velocardiofacial syndrome, including one prior report of autoimmune hemolytic anemia and immune thrombocytopenia. The patient reported herein strengthens the association between the 22q11.2 deletion spectrum and Evans syndrome. PMID- 12554530 TI - Uncompacted myelin lamellae in peripheral nerve biopsy. AB - Since 1979, the authors have studied 49 peripheral nerve biopsies presenting uncompacted myelin lamellae (UML). Based on the ultrastructural pattern of UML they propose a 3-category classification. The first category includes cases displaying regular UML, which was observed in 43 cases; it was more frequent in 9 cases with polyneuropathy organomegaly endocrinopathy m-protein skin changes (POEMS) syndrome as well as in 1 case of Charcot-Marie-Tooth 1B with a novel point mutation in the P0 gene. The second category consists of cases showing irregular UML, observed in 4 cases with IgM monoclonal gammopathy and anti-myelin associated glycoprotein (MAG) activity. This group included 1 benign case and 3 B cell malignant lymphomas. The third category is complex UML, which was present in 2 unrelated patients with an Arg 98 His missense mutation in the P0 protein gene. Irregular and complex UML are respectively related to MAG and P0, which play a crucial role in myelin lamellae compaction and adhesion. PMID- 12554531 TI - Myofibrosarcoma (low-grade myofibroblastic sarcoma) with intracytoplasmic hyaline (fibroma-like) inclusion bodies. AB - Myofibrosarcoma is a controversial neoplasm composed of cells with differentiation toward myofibroblasts. The authors report an unusual case of myofibrosarcoma in which, in addition to the characteristic features reported, tumor cells contained intracytoplasmic hyaline (fibroma-like) inclusion bodies. A 66-year-old man complained of a painless enlarged mass in his right shoulder over the previous 6 months. The tumor recurred 2 years after operation. On histologic examination, the tumor displayed diffusely infiltrative growth with isolation of individual skeletal muscle fibers. The tumor cells were most often arranged in an intersecting (herringbone) fascicular pattern but a vaguely storiform pattern was also observed. Cellularity varied from one area to another. The cells were spindle-shaped, with ill-define pale eosinophilic cytoplasm, and a wavy or tapering nucleus with fine chromatin and small nucleoli. Intracytoplasmic hyaline inclusion bodies of variable size, often located adjacent to the nucleus, were visible in numerous spindle and stellate cells. The mitotic index was 3/10 HPF. The tumor was scored as grade 1. Immunohistochemistry revealed positive staining to muscle-specific actin, desmin and vimentin in most tumor cells. Intracytoplasmic hyaline inclusion bodies did not react specifically to any one antibody, but showed a marked ring-like immune reaction, particularly to muscle specific actin. Electron microscopy showed tumor cells with indented nuclei and small nucleoli, abundant rough endoplasmic reticulum, micropinocytotic vesicles, and longitudinally arranged fine filaments with focal electron-dense patches and subplasmalemmal plaques. The most striking feature was the presence of large, globular or ball-like, non-membrane-bound, randomly scattered clusters of fine filament, usually adjacent to the nucleus. PMID- 12554532 TI - Ultrastructural changes and olfactory deficits during 3-methylindole-induced olfactory mucosal necrosis and repair in mice. AB - Olfactory mucosa from C57BL/6N mice was examined by transmission electron microscopy at 0.5, 1, 2, 4, 6, 24, 48, and 72 h after 400 mg 3-methylindole (3MI)/kg ip, and at 7, 14, and 21 days after 300 mg 3MI/kg or vehicle. Degeneration was evident in epithelial cells of Bowman's glands and olfactory sustentacular cells by 0.5 h, but not in neurons until 24 h, by which time necrosis was fully developed in sustentacular cells and epithelium of Bowman's glands. Sustentacular cells and neurons detached from the basal lamina by 48 h. Lamina proprial fibroblasts were hypertrophied by 72 h and, with collagen fibrils, formed the bulk of the mucosa at 7 days. By 21 days, fibroblasts were less conspicuous, but Bowman's glands were rarely observed and were lined by epithelial cells without secretory granules. Mucosal epithelium was reconstituted, but disorderly and lacked olfactory differentiation. Mice treated with 3MI were more likely than control mice to taste water treated with isoamyl acetate (odorant) and quinine monohydrochloride (aversive tastant). Though olfactory neurons were initially spared, their absence in the regenerated epithelium explains lingering olfactory deficits in murine 3MI toxicosis. PMID- 12554533 TI - The influence of different cultivating conditions on polymorphonuclear leukocyte apoptotic processes in vitro, I: the morphological characteristics of PMN spontaneous apoptosis. AB - Polymorphonuclear leukocyte (PMN) populations incubated in vitro with normal human serum are save-regulated systems of spontaneous apoptosis. Light microscopy (LM), transmission (TEM), and scanning (SEM) electron microscopes were used for the evalution of PMN apoptopic alteration. Twelve-hour PMN populations were represented by optimal number of normal and different apoptotic forms. Their ultrastructural analysis showed that on this background, 3 apoptotic cell lines (code named "first," "second," and "third") were predominated. The following characteristics were featured: "first"--vacuolization of same organelles, release of their content outside, increase of general cytoplasmic density, nuclear filling with condensed chromatin, and formation of PMNs mainly into small, round, dense forms; "second"--involvement of micronuclei or nuclei in apoptosis, their displacement to the cytoplasmic membrane and separation from the cells, and cytoplasm had numerous intact granules almost until the completion of apoptosis; "third"--synchronous apoptotic process of the nuclei and cytoplasm, moderate electronic density of cytoplasm, and granular translocation to the cell surface. Secondary necrosis was completed mainly in the apoptotic process of the "second" and "third" lines. SEM surfaces confirmed the results of TEM. This research showed that neutrophil spontaneous apoptosis is a complicated process. The 3 apoptotic cell lines reflect different pathways characteristic for the studied systems under certain conditions of cultivation. PMID- 12554534 TI - Role of the estrogen antagonist ICI 182,780 in vessel assembly and apoptosis of endothelial cells. AB - Angiogenesis is required during tumor progression. Emerging data, including the presence of estrogen receptors in endothelium, suggests that estrogens can mediate endothelial proliferation and differentiation. Therefore, it is likely that anti-estrogenic drugs can also exert their effects in endothelial cells. The purpose of this work was to evaluate the effect of one anti-estrogenic agent, ICI 182,780, in human umbilical vein endothelial cells (HUVECs). Treatment of HUVECs with 5 different concentrations of ICI 182,780 resulted in decreased cell viability and increase in apoptosis. Gene expression profile of these ICI-treated cells evaluated by cDNA array presented an upregulation of 68 newly expressed genes, whose expression was absent from both control and 17beta-estradiol-treated HUVECs. Most of these genes were implicated in both intrinsic and extrinsic apoptotic pathways. Furthermore, ICI 182,780 incubation prevented HUVECs from formity capillary-like tubules in a Matrigel assay. These findings suggest that besides blocking tumor cell proliferation in an estrogen receptor-dependent manner, ICI 182,780 impaired angiogenesis by preventing branching and capillary like tubule formation and by activating apoptotic pathways in endothelial cells. PMID- 12554535 TI - Vascular invasion in thyroid and gastric carcinomas. AB - The molecular, biological, and histopathologic features related to vascular invasiveness of gastric and thyroid carcinomas, with an emphasis on glandular (intestinal) carcinoma of the stomach and follicular thyroid carcinoma, are reviewed from a clinicopathologic standpoint. PMID- 12554537 TI - Pulmonary large cell carcinoma with rhabdoid phenotype. AB - A 70-year-old woman presented with a coin lesion in her left lung. The tumor was well circumscribed and had a large area of central necrosis with a thin rim of viable tumor cells. It showed a solid growth pattern of polygonal cells with eosinophilic intracytoplasmic inclusion bodies. Immunohistochemically, the tumor cells were positive for vimentin, neural cell adhesion molecule, neuron-specific enolase, and vascular endothelial growth factor. Electron microscopy revealed intracytoplasmic inclusion bodies consisting of whorled intermediate filaments. Based on histological and immunohistochemical findings, the patient was diagnosed as having pulmonary large cell carcinoma with rhabdoid phenotype (LCCRP). The patient was in stage IA, and the histological findings may be the prototype of pure LCCRP. The tumor recurred after 6 years, and the second tumor had more apparent intracytoplasmic inclusion bodies. It is worthwhile detecting and recognizing the significance of these intracytoplasmic inclusions because of the poor prognosis of this tumor. PMID- 12554536 TI - Curvilinear membranous formations in lymph node cells of an African AIDS patient with tuberculosis. AB - Tissue from an extirpated cervical lymph node of an African AIDS patient with tuberculosis was investigated with the electron microscope. A peculiar cytoplasmic structure was found in histiocytic cells of the lymph node tissue. This structure consisted of a loose aggregation of convoluted, folded curvilinear membranes, demonstrating C-shaped, U-shaped, and S-shaped profiles according to the plane of section. These curvilinear membranous formations (CMF) were formed by 2 parallel-running unit membranes separated by a narrow electron-lucent space. Continuities of CMF with rough-surfaced endoplasmic reticulum were sometimes observed. Occasionally, an electron-dense, blurred cytoplasmic area was seen between 2 neighboring CMF. Several CMF also formed a seastar-like configuration by mergence in such an area. It is unknown whether the HIV infection of the patient was the primary cause of the CMF. PMID- 12554538 TI - BK polyomavirus interstitial nephritis in a renal allograft recipient. AB - Human polyomavirus (PV) interstitial nephritis has recently been recognized as a cause of severe renal allograft dysfunction. It occurs in immunosuppressed patients after reactivation of the latent virus PV type BK (BK virus) in the renal epithelium. BK disease is defined as a morphologically manifest renal infection with cytopathic signs accompanied by varying degrees of interstitial inflammatory cell infiltrates and functional impairment. It is also identified by the presence of cells containing viral inclusion bodies (decoy cells) in the urine. The authors report a case of BK PV interstitial nephritis in a 36-year-old renal allograft recipient. Under light microscopy the chief diagnostic indicator was detection of intranuclear viral inclusions, which were found exclusively in tubular epithelial cells. Cells with viral changes were often enlarged with nuclear atypia and chromatin basophilia. Widespread interstitial plasma cell infiltrates associated with tubulitis were present. Intranuclear paracrystalline arrays of virus particles 35-38 nm in diameter were present as characteristic ultrastructural indicators. Urine samples revealed decoy cells with ground-glass type intranuclear inclusions positive for BK virus by electron microscopy. PMID- 12554539 TI - Ecotoxicological risks associated with land treatment of petrochemical wastes. I. Residual soil contamination and bioaccumulation by cotton rats (Sigmodon hispidus). AB - Petrochemical waste contains both organic and inorganic contaminants that can pollute soil and may pose significant ecological risks to wildlife. Petrochemical waste typically is disposed of in land treatment units, which are widespread throughout Oklahoma and the United States. Few studies have been conducted evaluating possible toxicity risks to terrestrial organisms residing on these units. In this study, the extent of soil contamination with fluoride (F), metals, and organic hydrocarbons, the bioaccumulation of F and metals in cotton rats (Sigmodon hispidus), the relationship between contaminants in soil and in tissues of cotton rats, and the level of potentially toxic polycyclic aromatic hydrocarbons (PAHs) in soil were determined on land treatment units. Over a 2-yr period, cotton rats and soils were collected and analyzed from 5 land treatment and matched reference units. The number of land treatment units with soil metal contamination (in parentheses) included: Cr, Cu, Pb (5). Al, As, Ni, Sr, Zn (4). Ba (3). and Cd, V (2). The number of land treatment units with soil PAH contamination (in parentheses) were naphthalene, phenanthrene, benzo[g,h,i]perylene (3). acenaphthene, anthracene, pyrene, benz[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[a]pyrene, indeno[1,2,3-c,d]pyrene, dibenz[a,h]anthracene (2). and acenaphthylene, fluorene, fluoranthene, benzo[k]fluoranthene (1). Total PAH and total petroleum hydrocarbons (TPH) were elevated at all five land treatment units. Mean sums of benzo[a]pyrene (BaP) equivalents (BaPequiv ) were not affected on PMID- 12554541 TI - Ecotoxicological risks associated with land treatment of petrochemical wastes. III. Immune function and hematology of cotton rats. AB - Landfarming is a widely used method of treating petrochemical waste through microbial bio-degradation. The effects of residual petrochemical contamination on wildlife, especially terrestrial mammals, are poorly understood. The effects of contaminants on the immune system and hematology of cotton rats (Sigmodon hispidus) living on five abandoned petrochemical landfarms (units 1-5) in Oklahoma were studied. Cotton rats were sampled seasonally (summer and winter) from each landfarm and from five ecologically matched reference sites for 2 yr (1998-2000) and returned to the laboratory for immunological and hematological assays. Overall analysis indicated that rats inhabiting landfarms exhibited decreased relative spleen size compared to rats collected from reference sites, with the landfarm at unit 1 showing the greatest reduction. Cotton rats collected from landfarms also had increased hemoglobin, hematocrit, and platelet levels and decreased blood leukocytes during summer. During winter, an increase in the number of popliteal node white blood cells was observed from rats collected on landfarms. No marked difference was detected for lymphocyte proliferation in response to concanavalin A, pokeweed, or interleukin-2. Lymphokine-activated killer cell lytic ability showed a seasonal pattern, but no treatment differences. No differences between landfarm and reference sites were detected in the hypersensitivity reaction of rats given an intradermal injection of phytohemagluttinin (PHA-P). Comparisons within individual sites indicated that two sites (units 1 and 3) had the greatest effects on immune function and hematology of cotton rats. The results of this study suggest that residual petrochemical waste affects the immune system and hematology of cotton rats living on abandoned landfarms during summer and is complicated by variation in the contaminants found on individual petroleum sites. PMID- 12554540 TI - Ecotoxicological risks associated with land treatment of petrochemical wastes. II. Effects on hepatic phase I and phase II detoxification enzymes in cotton rats. AB - The purpose of this study was to evaluate possible exposure and resultant hepatic effects of petrochemical waste on cotton rats (Sigmodon hispidus) living on landfarmed sites. Male and female cotton rats were collected in summer, fall, and winter from four landfarm sites and four ecologically similar reference sites. Hepatic methoxyresorufin O-deethylase (MROD) activity was significantly induced in male and female rats collected from landfarms compared to rats collected from reference sites. In contrast, changes in ethoxyresorufin O-deethylase (EROD) activity were inconsistent due to season, sex, and treatment variation. A significant decrease in EROD and MROD activity was found in cotton rats held for 48 h prior to sacrifice compared to rats euthanized on the day of capture. These results indicate that when using hepatic EROD and MROD activities as biochemical markers of exposure to aryl hydrocarbon receptor agonists, animals should be euthanized as quickly as possible after capture. The cotton rats collected from one landfarm unit exhibited a pattern of consistent elevation of EROD, MROD, and pent-oxyresorufin O-deethylase (PROD) activity. This unit also had a pattern of elevated CYP1A2 protein expression determined by Western blotting. There were no consistent alterations from contaminant exposure on hepatic glutathione S transferase (GST) activity, glutathione levels, or CYP1A1 protein. Hepatic EROD and MROD activities varied considerably between seasons and sex of rats. In conclusion, consistent induction of hepatic EROD and MROD activities in cotton rats was found in three out of four sampled landfarm sites compared to the rats collected from the reference sites, indicating exposure to contaminants-likely polyaromatic hydrocarbons. PMID- 12554542 TI - Magnetometric evaluation of cadmium oxide-induced toxicity to pulmonary alveolar macrophages of Syrian golden hamsters. AB - Since alveolar macrophages play an important role in the clearance of inhaled dust from air-ways, these cells have been used as a target for various toxic chemicals. Alveolar macrophages obtained from bronchoalveolar lavage of Syrian golden hamsters were concurrently exposed in vitro to Fe(3)O(4), as an indicator for magnetometry, and various concentrations of cadmium oxide (CdO) in this study. A rapid decrease of the remnant magnetic field, called relaxation, was observed after the cessation of an external magnetic field stimulus in macrophages concurrently exposed to phosphate-buffered saline or CdO at 0.1 microg/ml, while relaxation was delayed in those concurrently exposed to 1, 25, or 50 microg/ml CdO. Therefore, the concentration of CdO affecting relaxation in vitro was estimated at between 0.1 and 1 microg/ml. Release of LDH activity from CdO-exposed macrophages into the medium significantly increased at levels of 25 and 50 microg/ml CdO. Apoptosis was not detected in macrophages exposed to CdO by the DNA ladder detection method or morphological observations. Electron microscopic examination revealed severe membrane damage and vacuolar changes in macrophages exposed to CdO. Since delayed relaxation is thought to occur by (1). disrupted cytoskeleton-driven random rotation of phagosomes containing iron oxide particles, (2). significant lactate dehydrogenase (LDH) activity release, and (3). detachment of cell membranes, CdO is considered to affect macrophage functions. PMID- 12554543 TI - Modulation of lipopolysaccharide-induced proinflammatory cytokine production by satratoxins and other macrocyclic trichothecenes in the murine macrophage. AB - The satratoxins and other macrocyclic trichothecene mycotoxins are produced by Stachybotrys, a mold that is often found in water-damaged dwellings and office buildings. To test the potential immunomodulatory effects of these mycotoxins, RAW 264.7 murine macrophage cells were treated with various concentrations of satratoxin G (SG), isosatratoxin F (iSF), satratoxin H (SH), roridin A (RA), and verrucarin A (VA) for 48 h in the presence or absence of suboptimal concentra tion of lipopolysaccharide (LPS, 50 ng/ml), and tumor necrosis factor-alpha (TNF alpha ) and interleukin-6 (IL-6) production were assayed by enzyme-linked immunosorbent assay (ELISA). In LPS-stimulated cultures, TNF-alpha supernatant concentrations were significantly increased in the presence of 2.5, 2.5, and 1 ng/ml of SG, SH, and RA, respectively, whereas IL-6 concentrations were not affected by the same concentrations these macrocyclic trichothecenes. When cells that were treated with LPS and SG (2.5 ng/ml) were evaluated by real-time polymerase chain reaction (PCR),TNF-alpha mRNA was found to increase at 24, 36, and 48 h compared to control cells. At higher concentrations, cytokine production and cell viability were markedly impaired in LPS-stimulated cells. Without LPS stimulation, neither TNF-alpha, nor IL-6 was induced. These results indicate that low concentrations of macrocyclic trichothecenes superinduce expression of TNF alpha, whereas higher concentrations of these toxins are cytotoxic and concurrently reduce cytokine production. The capacity of satratoxins and other macrocyclic trichothecenes to alter cytokine production may play an etiologic role in outbreaks of Stachybotrys-associated human illnesses. PMID- 12554544 TI - Kinetics of an intratracheally administered chromium catalyst in rats. AB - Chromium-based catalysts are used for the synthesis of polyethylene, but little is known about the hazard and biomonitoring possibilities of this type of chromium for workers who may be occupationally exposed to such compounds. Therefore, the bioavailability and toxicokinetics of chromium were studied in male Wistar rats after a single intratracheal instillation (2 ml/kg body weight) of various doses (1, 5, or 25 mg/kg body weight) of the catalyst (approximately 1% chromium bound to an amorphous silica matrix), either before (CAT-Cr[III]) or after (CAT-Cr[VI]) heat treatment. The results were compared with those of equivalent amounts of two chromium salts (CrCl(3) and K(2) Cr(2) O (7). Each dose group was composed of three rats. The concentration of chromium was determined by atomic absorption spectrometry in urine (collected daily for 7 d) and in plasma, erythrocytes, lung, and liver tissue obtained 2 d (only highest concentration) and 7 d after dosing. On d 2, a significant increase in lung weight was found in the animals treated with the highest dose of the hexavalent Cr products. On d 7, on the basis of body weights, lung weights, and lung histology, there was no overt toxicity, except after the highest dose of CAT-Cr(VI). The elimination of all forms of chromium was apparently monoexponential, with calculated half-life elimination times in urine of 4-11 h for Cr(III) (CAT-Cr[III] and CrCl3 ) and 8 21 h for Cr(VI) (CAT-Cr[VI] and K(2) Cr(2) O(7). On d 2, the erythro-cytes Cr concentrations were significantly higher for the hexavalent Cr products than for the trivalent Cr products. After 7 d, the erythrocytes Cr concentrations were significantly increased above control values (3 microg/L) only in rats treated with the 2 highest doses of Cr( VI) compounds (12 and 64 microg/L for K(2) Cr(2) O(7), and 14 and 79 microg/L for CAT-Cr[VI]). The present study shows that intratracheally instilled Cr(VI) and Cr(III) have different toxicokinetic profiles and that the Cr(VI) catalyst has the same bioavailability and excretion kinetics as a water-soluble Cr(VI) salt. Exposure to chromium compounds could be monitored by measuring Cr concen-trations in urine (shortly after exposure) and in erythrocytes (also at later time points after high Cr[VI] exposure). PMID- 12554545 TI - Overcoming depression on the Internet (ODIN): a randomized controlled trial of an Internet depression skills intervention program. AB - BACKGROUND: Psychoeducational programs are increasingly being delivered over the Internet. We created an Internet-based, cognitive therapy, self-help program to be used as a stand-alone intervention for mild-to-moderate depression, or as an adjunct to traditional services for more severe depression. OBJECTIVE: To evaluate the effectiveness of a web-based intervention program to reduce depression in a randomized, controlled trial. METHODS: In a private, nonprofit health maintenance organization, we mailed recruitment brochures to two populations: depressed adults (n = 6994) who received traditional medical services for depression, and an age/gender matched sample of nondepressed adults (n = 6996). Participants consenting to the study were randomized to either the experimental Web site (n = 144), or a no-access control group (n = 155). Participants in both groups were free to obtain nonexperimental, usual care services for their depression. All participants completed an on-line version of the Center for Epidemiological Studies Depression Scale (CES-D) at enrollment and at 4-, 8-, 16- and 32-weeks after enrollment. Mean intake scores were in the severely depressed range. 74% of participants completed at least one follow-up assessment. Unfortunately, most intervention participants accessed the Internet site infrequently. RESULTS: We failed to find an effect for the Internet program across the entire sample. However, post-hoc, exploratory analyses revealed a modest effect among persons reporting low levels of depression at intake. CONCLUSIONS: The negative results might have resulted from infrequent patient use of the Internet site, or a more seriously depressed sample than the intervention was intended to help. Future studies should focus on recruiting persons with mild to moderate levels of depression, and on increasing participant use of the Internet site. PMID- 12554546 TI - eEurope 2002: Quality Criteria for Health Related Websites. AB - BACKGROUND: A number of organisations have begun to provide specific tools for searching, rating, and grading this information, while others have set up codes of conduct by which site providers can attest to their high quality services. The aim of such tools is to assist individuals to sift through the mountains of information available so as to be better able to discern valid and reliable messages from those which are misleading or inaccurate. OBJECTIVE: Recognising that European citizens are avid consumers of health related information on the internet and recognising that they are already using the types of rating system described above, the European Council at Feira on June 19-20 2000 supported an initiative within eEurope 2002 to develop a core set of Quality Criteria for Health Related Websites. The specific aim was to draw up a commonly agreed set of simple quality criteria on which Member States, as well as public and private bodies, may draw in the development of quality initiatives for health related websites. These criteria should be applied in addition to relevant Community law. METHODS: A meeting was held during 2001 which drew together key players from Government departments, International Organisations, non-governmental organisations and industry, to explore current practices and experiments in this field. Some sixty invited participants from all the Member States, Norway, Switzerland, and the United States of America took part in the meeting of June 7 8, 2001: they included delegates from industrial, medical, and patient interest groups, delegates from Member States' governments, and key invited speakers from the field of health information ethics. These individuals, and many others, also took part in the web-based consultation which was open from august to November 2001. RESULTS: The broad headings for quality criteria identified include Transparency and Honesty, Authority, Privacy and data protection, Updating of information, Accountability, Responsible partnering, Editorial policy, Accessibility, the latter includes attention to guidelines on physical accessibility as well as general findability, searchability, readability, usability, etc. A metadata labelling system may be used to make health data more findable. Such a system may also be used in conjunction with quality criteria to give higher ranking by search engines to those sites or pages labelled as complying with defined quality criteria. CONCLUSIONS: The set of quality criteria is based upon a broad consensus among specialists in this field, health authorities, and prospective users. It is now to be expected that national and regional health authorities, relevant professional associations, and private medical website owners will 1) implement the Quality Criteria for Health Related Websites in a manner appropriate to their website and consumers; 2) develop information campaigns to educate site developers and citizens about minimum quality standards for health related websites; 3) draw on the wide range of health information offered across the European Union and localise such information for the benefit of citizens (translation and cultural adaptation); 4) exchange information and experience at European level about how quality standards are being implemented. PMID- 12554547 TI - Evaluation of accessibility and use of new communication technologies in patients with type 1 diabetes mellitus. AB - BACKGROUND: The role of patients in the management and control of type 1 diabetes mellitus, a chronic disease, is well established. The advent of new communication technologies is expected to improve patients' access to health information. However, little is known about the extent to which patients with type 1 diabetes mellitus use the Internet to retrieve medical information and about the impact, if any, this retrieval has on their health status. OBJECTIVE: To evaluate the accessibility and use of new communication technologies in a population of patients with type 1 diabetes mellitus. METHODS: Patients with type 1 diabetes mellitus attending the Diabetes Clinic of the Hospital de Sabadell, Sabadell, Spain, in a 6-month period were asked to answer a structured questionnaire about education level, Internet accessibility, use of health-related Web sites, and mobile-phone ownership and use. RESULTS: Of 302 patients with type 1 diabetes mellitus attending the Diabetes Clinic on a regular basis, 244 (115 men, 129 women) were interviewed (response rate 80.8%). Personal computers were owned by 58.2% of patients. Fifty-nine percent had access to the Internet, 39.3% had access to the Internet at home; however, only 36.5% were regular Internet users. Internet users were younger, more frequently men, and of higher education level. Among Internet users only 49.4% had ever accessed a health-related Web site. Internet users who had ever accessed a health-related Web site had a higher level of education, presented severe hypoglycemia more frequently, and were more likely to have access to the Internet at home. No differences were found in metabolic control between Internet users and nonusers or between Internet users who had ever accessed a health-related Web site and Internet users who had never accessed a health-related Web site. Of the 76.6% of the patients that owned a mobile phone, 96% used it more than once a week. CONCLUSIONS: The impact of new communication technologies might be jeopardized by the low rate of access and utilization of the Internet for health-related purposes. Because of their high rate of ownership and use, mobile phones show promise as a tool in health care communication technologies. PMID- 12554548 TI - Issues in evaluating health websites in an Internet-based randomized controlled trial. PMID- 12554549 TI - Asphyxial death by ether inhalation and plastic-bag suffocation instructed by the press and the Internet. PMID- 12554550 TI - Unmet needs of primary care patients in using the Internet for health-related activities. AB - BACKGROUND: Millions of people use the Internet as a source for health information yet little is understood about the use of the Internet for other health-related activities. OBJECTIVE: We conducted the present study to understand, among primary care patients, the interest in and experience with using the Internet for a variety of health-related activities. METHODS: Cross sectional survey in the setting of 4 community-based primary care practices in Rhode Island. A single self-administered questionnaire included the following: 14 items measuring interest in using the Internet for a variety of health-related purposes, demographics, self-reported health status, and self-reported health care quality. RESULTS: The survey was completed by 300 patients, 109 without access to the Internet and 191 with access to the Internet. Experiences with and attitudes about each of the health-related activities on the Internet varied widely across each activity. Regardless of access, patients were most interested in using the Internet for finding information about diseases and medications. However, patients with Internet access were more interested, compared to those without access, in each of the health-related activities on the Internet. Among patients with access to the Internet, the largest gap between interest and experience (the opportunity gap) was in using the Internet to investigate the quality of their care (eg, "find out if your health care provider was giving you all of the tests and treatments that you are due to have?") and administrative functions (eg, "schedule an appointment with your doctor?"). CONCLUSIONS: Much opportunity remains for developing health-related Internet Web sites to address the unmet needs of primary care patients. PMID- 12554551 TI - Implementation and integration of regional health care data networks in the Hellenic National Health Service. AB - BACKGROUND: Modern health care is provided with close cooperation among many different institutions and professionals, using their specialized expertise in a common effort to deliver best-quality and, at the same time, cost-effective services. Within this context of the growing need for information exchange, the demand for realization of data networks interconnecting various health care institutions at a regional level, as well as a national level, has become a practical necessity. OBJECTIVES: To present the technical solution that is under consideration for implementing and interconnecting regional health care data networks in the Hellenic National Health System. METHODS: The most critical requirements for deploying such a regional health care data network were identified as: fast implementation, security, quality of service, availability, performance, and technical support. RESULTS: The solution proposed is the use of proper virtual private network technologies for implementing functionally interconnected regional health care data networks. CONCLUSIONS: The regional health care data network is considered to be a critical infrastructure for further development and penetration of information and communication technologies in the Hellenic National Health System. Therefore, a technical approach was planned, in order to have a fast cost-effective implementation, conforming to certain specifications. PMID- 12554552 TI - Internet infrastructures and health care systems: a qualitative comparative analysis on networks and markets in the British National Health Service and Kaiser Permanente. AB - BACKGROUND: The Internet and emergent telecommunications infrastructures are transforming the future of health care management. The costs of health care delivery systems, products, and services continue to rise everywhere, but performance of health care delivery is associated with institutional and ideological considerations as well as availability of financial and technological resources. OBJECTIVE: to identify the effects of ideological differences on health care market infrastructures including the Internet and telecommunications technologies by a comparative case analysis of two large health care organizations: the British National Health Service and the California-based Kaiser Permanente health maintenance organization. METHODS: A qualitative comparative analysis focusing on the British National Health Service and the Kaiser Permanente health maintenance organization to show how system infrastructures vary according to market dynamics dominated by health care institutions ("push") or by consumer demand ("pull"). System control mechanisms may be technologically embedded, institutional, or behavioral. RESULTS: The analysis suggests that telecommunications technologies and the Internet may contribute significantly to health care system performance in a context of ideological diversity. CONCLUSIONS: The study offers evidence to validate alternative models of health care governance: the national constitution model, and the enterprise business contract model. This evidence also suggests important questions for health care policy makers as well as researchers in telecommunications, organizational theory, and health care management. PMID- 12554553 TI - Internet use in disease management for home care patients: a call for papers. PMID- 12554554 TI - Users of Internet health information: differences by health status. AB - BACKGROUND: Millions of consumers have accessed health information online. However, little is known about their health status. OBJECTIVE: To explore use of Internet health information among those who were sicker (fair/poor general health status) compared with those reported being healthier. METHODS: A national, random digit telephone survey by the Pew Internet & American Life Project identified 521 Internet users who go online for health care information. Our primary independent variable was general health status rated as excellent, good, fair, or poor. Patterns of Internet use, and types of information searched were assessed. RESULTS: Among the 521 users, 64% were female, most (87%) were white, and median age was 42 years. Most individuals indicated that they learned something new online (81%) and indicated that they believe most information on the Internet (52%). Compared with those with excellent/good health, those with fair/poor health (N = 59) were relative newcomers to the Internet but tended to use the Internet more frequently, were more likely to use online chats, were less likely to search for someone other than themselves, and were more likely to talk about the new information with their physician (odds ratio 3.3 [95% confidence interval 1.8-6.3]), after adjustment for age, education and income. CONCLUSIONS: Health care professionals should be aware that their sicker patients are more likely to ask them about information they found online. Physicians, public health professionals, and eHealth developers should work together to educate patients about searching for health information online and to provide tools for them to navigate to the highest quality information. PMID- 12554555 TI - Smoking cessation and the Internet: a qualitative method examining online consumer behavior. AB - BACKGROUND: Smoking is a major preventable cause of disease and disability around the world. Smoking cessation support-including information, discussion groups, cognitive behavioral treatment, and self-help materials-can be delivered via the Internet. There is limited information about the reasons and methods consumers access smoking cessation information on the Internet. OBJECTIVES: This study aims to determine the feasibility of a method to examine the online behavior of consumers seeking smoking cessation resources. In particular, we sought to identify the reasons and methods consumers use to access and assess the quality of these resources. METHODS: Thirteen participants were recruited via the state based Quit smoking cessation campaign, operated by the Victorian Cancer Council, in December 2001. Online behavior was evaluated using semi-structured interviews and Internet simulations where participants sought smoking cessation information and addressed set-case scenarios. Online interaction was tracked through pervasive logging with specialist software. RESULTS: Thirteen semi-structured interviews and 4 Internet simulations were conducted in January 2002. Participants sought online smoking cessation resources for reasons of convenience, timeliness, and anonymity-and because their current information needs were unmet. They employed simple search strategies and could not always find information in an efficient manner. Participants employed several different strategies to assess the quality of online health resources. CONCLUSIONS: Consumer online behavior can be studied using a combination of survey, observation, and online surveillance. However, further qualitative and observational research is required to harness the full potential of the Internet to deliver public health resources. PMID- 12554556 TI - Use of the Internet by women with breast cancer. AB - BACKGROUND: Recently, many cancer patients have been using the Internet for information with which to make informed choices. We are not aware of any studies that investigate this Internet use among breast cancer patients or women. OBJECTIVE: We investigate the prevalence and predictors of Internet use for medical information among women with breast cancer. METHODS: We used a cross sectional design and approached 251 women with breast cancer being treated at a university-based hospital. We successfully interviewed 188 (74.9%), through mailed self-report questionnaires. Medical information was obtained from the hospital tumor registry. We used t tests and chi-square tests to assess differences in Internet use for breast health issues and binary logistic regression to estimate the odds ratio (OR) for predictors of Internet use for breast health issues. RESULTS: In our sample, 41.5% of patients used the Internet for medical information. Internet users differed from nonusers on income level, educational level, and by race/ethnicity. After controlling for the other predictors, Internet users had a higher income (OR = 3.10; 95% CI = 1.09-8.85) and tended to be more educated (OR = 2.59; 95% CI = 0.87-7.74) than nonusers. There was also a suggestion that those of nonwhite ethnicity were less likely to use the Internet (OR = 0.39; 95% CI = 0.14-1.11). Increasing age, length of time since diagnosis, and breast cancer stage had no effect. CONCLUSIONS: A substantial proportion of breast cancer patients used the Internet as a source of information. Patients with higher income or education, and patients of white race/ethnicity are more likely to use the Internet for breast health issues. PMID- 12554557 TI - Where are they now? A case study of health-related Web site attrition. AB - BACKGROUND: When considering health-related Web sites, issues of quality generally focus on Web content. Little concern has been given to attrition of Web sites or the "fleeting" nature of health information on the World Wide Web. Since Web sites may be available for an uncertain period of time, a Web page may not be a sound reference. OBJECTIVE: To address the issue of attrition, a defined set of health-related Web sites was examined at two separate time intervals. METHODS: To determine the degree of attrition, Web sites obtained and recorded from a previous study were revisited approximately three years later. From December 1998 to May 1999, 184 Web sites were collected from which health claims were identified. During May and June 2002, the previously recorded URL for each Web site was entered into the address field of the browser Netscape Navigator. It was documented whether the original Web site could not be found, moved to a different URL location, or the URL and site location was found unchanged from the original search. For a Web site whose URL remained unchanged, it was also noted whether the site had maintained currency, (i.e. updated) since the original posting. To ensure that inaccessibility may not be due to temporary server problems, another attempt was made to access the sites at different periods of time. RESULTS: When each URL address from the original set of 184 Web sites was re-entered into the address field of the browser, 108 (59%) of the sites could not be found, 31 (17%) had moved to a new URL address, and 45 (24%) of the sites could be found from the original URLs obtained in the previous study. Of the Web sites that moved to a new URL address, 7 sites provided a link from the original URL to redirect the viewer to the new location. Of the Web sites still in existence, 17 (38%) provided update information from the original posting. CONCLUSIONS. It can be difficult to locate information that was previously found on the Web, and if a reference to an item is provided, there is no guarantee that viewers will be able to find the site at a later time. Enhancements in Web technologies such as the Internet Archive may improve this situation. Future research that is directed toward making sure Web site viewers know the site will be accessible at a later time will enhance the Web as a valuable medical information resource. PMID- 12554558 TI - A review of features in Internet consumer health decision-support tools. AB - BACKGROUND: Over the past decade, health care consumers have begun to benefit from new Web-based communications tools to guide decision making on treatments and tests. Using today's online tools, consumers who have Internet connections can: watch and listen to videos of physicians; watch and hear the stories of other consumers who have faced the same decisions; join an online social support network; receive estimates of their own chances of experiencing various outcomes; and do it all at home. OBJECTIVE: To review currently-available Internet consumer health decision-support tools. METHODS: Five Web sites offering consumer health decision-support tools are analyzed for their use of 4 key Web-enabled features: the presentation of outcomes probability data tailored to the individual user; the use of videotaped patient interviews in the final product to convey the experiences of people who have faced similar diagnoses in the past; the ability to interact with others in a social support network; and the accessibility of the tool to any health care consumers with an Internet connection. RESULTS: None of the 5 Web sites delivers all 4 target features to all Web users. The reasons for these variations in the use of key Web functionality--features that make the Web distinctive--are not immediately clear. CONCLUSIONS: Consumers trying to make health care decisions may benefit from current Web-based decision-support tools. But, variations in Web developers' use of 4 key Web-enabled features leaves the online decision-support experience less than what it could be. Key research questions are identified that could help in the development of new hybrid patient decision-support tools. PMID- 12554559 TI - Security, privacy, and confidentiality issues on the Internet. AB - We introduce the issues around protecting information about patients and related data sent via the Internet. We begin by reviewing three concepts necessary to any discussion about data security in a healthcare environment: privacy, confidentiality, and consent. We are giving some advice on how to protect local data. Authentication and privacy of e-mail via encryption is offered by Pretty Good Privacy (PGP) and Secure Multipurpose Internet Mail Extensions (S/MIME). The de facto Internet standard for encrypting Web-based information interchanges is Secure Sockets Layer (SSL), more recently known as Transport Layer Security or TLS. There is a public key infrastructure process to 'sign' a message whereby the private key of an individual can be used to 'hash' the message. This can then be verified against the sender's public key. This ensures the data's authenticity and origin without conferring privacy, and is called a 'digital signature'. The best protection against viruses is not opening e-mails from unknown sources or those containing unusual message headers. PMID- 12554560 TI - Using the Internet for surveys and health research. AB - This paper concerns the use of the Internet in the research process, from identifying research issues through qualitative research, through using the Web for surveys and clinical trials, to pre-publishing and publishing research results. Material published on the Internet may be a valuable resource for researchers desiring to understand people and the social and cultural contexts within which they live outside of experimental settings, with due emphasis on the interpretations, experiences, and views of 'real world' people. Reviews of information posted by consumers on the Internet may help to identify health beliefs, common topics, motives, information, and emotional needs of patients, and point to areas where research is needed. The Internet can further be used for survey research. Internet-based surveys may be conducted by means of interactive interviews or by questionnaires designed for self-completion. Electronic one-to one interviews can be conducted via e-mail or using chat rooms. Questionnaires can be administered by e-mail (e.g. using mailing lists), by posting to newsgroups, and on the Web using fill-in forms. In "open" web-based surveys, selection bias occurs due to the non-representative nature of the Internet population, and (more importantly) through self-selection of participants, i.e. the non-representative nature of respondents, also called the 'volunteer effect'. A synopsis of important techniques and tips for implementing Web-based surveys is given. Ethical issues involved in any type of online research are discussed. Internet addresses for finding methods and protocols are provided. The Web is also being used to assist in the identification and conduction of clinical trials. For example, the web can be used by researchers doing a systematic review who are looking for unpublished trials. Finally, the web is used for two distinct types of electronic publication. Type 1 publication is unrefereed publication of protocols or work in progress (a 'post-publication' peer review process may take place), whereas Type 2 publication is peer-reviewed and will ordinarily take place in online journals. PMID- 12554562 TI - Spirituality and health. PMID- 12554564 TI - Avoidable mortality. PMID- 12554565 TI - Acupuncture and smoking cessation: pinning down the claims. PMID- 12554566 TI - Confronting the moral economy of US racial/ethnic health disparities. PMID- 12554567 TI - Investigating the role of racial/ethnic bias in health outcomes. PMID- 12554568 TI - A fresh approach to health care in the United States: improved and expanded Medicare for all. PMID- 12554569 TI - Does racism harm health? Did child abuse exist before 1962? On explicit questions, critical science, and current controversies: an ecosocial perspective. AB - Research on racism as a harmful determinant of population health is in its infancy. Explicitly naming a long-standing problem long recognized by those affected, this work has the potential to galvanize inquiry and action, much as the 1962 publication of the Kempe et al. scientific article on the "battered child syndrome" dramatically increased attention to-and prompted new research on the myriad consequences of child abuse, a known yet neglected social phenomenon. To further work on connections between racism and health, the author addresses 3 interrelated issues: (1) links between racism, biology, and health; (2) methodological controversies over how to study the impact of racism on health; and (3) debates over whether racism or class underlies racial/ethnic disparities in health. PMID- 12554570 TI - Racial/ethnic discrimination and health: findings from community studies. AB - The authors review the available empirical evidence from population-based studies of the association between perceptions of racial/ethnic discrimination and health. This research indicates that discrimination is associated with multiple indicators of poorer physical and, especially, mental health status. However, the extant research does not adequately address whether and how exposure to discrimination leads to increased risk of disease. Gaps in the literature include limitations linked to measurement of discrimination, research designs, and inattention to the way in which the association between discrimination and health unfolds over the life course. Research on stress points to important directions for the future assessment of discrimination and the testing of the underlying processes and mechanisms by which discrimination can lead to changes in health. PMID- 12554571 TI - Environmental equity and health: understanding complexity and moving forward. AB - The authors invoke a population health perspective to assess the distribution of environmental hazards according to race/ethnicity, social class, age, gender, and sexuality and the implications of these hazards for health. The unequal burden of environmental hazards borne by African American, Native American, Latino, and Asian American/Pacific Islander communities and their relationship to well documented racial/ethnic disparities in health have not been critically examined across all population groups, regions of the United States, and ages. The determinants of existing environmental inequities also require critical research attention. To ensure inclusiveness and fill important gaps, scientific evidence is needed on the health effects of the built environment as well as the natural environment, cities and suburbs as well as rural areas, and indoor as well as outdoor pollutants. PMID- 12554572 TI - Future directions in residential segregation and health research: a multilevel approach. AB - The authors examine the research evidence on the effect of residential segregation on health, identify research gaps, and propose new research directions. Four recommendations are made on the basis of a review of the sociological and social epidemiology literature on residential segregation: (1) develop multilevel research designs to examine the effects of individual, neighborhood, and metropolitan-area factors on health outcomes; (2) continue examining the health effects of residential segregation among African Americans but also initiate studies examining segregation among Hispanics and Asians; (3) consider racial/ethnic segregation along with income segregation and other metropolitan area factors such as poverty concentration and metropolitan governance fragmentation; and (4) develop better conceptual frameworks of the pathways that may link various segregation dimensions to specific health outcomes. PMID- 12554573 TI - Sick and tired of being sick and tired: scientific evidence, methods, and research implications for racial and ethnic disparities in occupational health. AB - The extent of racial/ethnic disparities in occupational health have not been well studied. The author reviews the evidence about workers of color and occupational injuries and disease. Patterns of employment in the U.S. workforce according to education, gender, and race/ethnicity are discussed, and how these patterns might cause disproportionate exposure leading to disproportionate disease and injury. Methodological issues are explored that have hampered research about occupational health disparities, and future research needs are identified. PMID- 12554574 TI - Employment discrimination, segregation, and health. AB - The author examines available evidence on the effects of exposure to joblessness on emotional well-being according to race and sex. The impact of racism on general health outcomes also is considered, particularly racism in the specific form of wage discrimination. Perceptions of racism and measured exposures to racism may be distinct triggers for adverse health outcomes. Whether the effects of racism are best evaluated on the basis of self-classification or social classification of racial identity is unclear. Some research sorts between the effects of race and socioeconomic status on health. The development of a new longitudinal database will facilitate more accurate identification of connections between racism and negative health effects. PMID- 12554575 TI - Perceived discrimination and depression: moderating effects of coping, acculturation, and ethnic support. AB - The authors evaluated the effects of cultural norms and social contexts on coping processes involved in dealing with perceived racial discrimination. Cross sectional data derived from personal interviews with Korean immigrants residing in Toronto were analyzed. Among the respondents, active, problem-focused coping styles were more effective in reducing the impacts on depression of perceived discrimination, while frequent use of passive, emotion-focused coping had debilitating mental health effects. The present findings lend greater support to a social contextual explanation than to a cultural maintenance explanation of coping processes. They also suggest that, when empowered with sufficient social resources, racial minority individuals of diverse cultural heritages are more likely to confront than to accept racial bias. PMID- 12554576 TI - Bias in mental health assessment and intervention: theory and evidence. AB - A recent surgeon general's report and various studies document racial and ethnic disparities in mental health care, including gaps in access, questionable diagnostic practices, and limited provision of optimum treatments. Bias is a little studied but viable explanation for these disparities. It is important to isolate bias from other barriers to high-quality mental health care and to understand bias at several levels (practitioner, practice network or program, and community). More research is needed that directly evaluates the contribution of particular forms of bias to disparities in the area of mental health care. PMID- 12554577 TI - Physiological responses to racism and discrimination: an assessment of the evidence. AB - A growing body of research explores the impact of encounters with racism or discrimination on physiological activity. Investigators have collected these data in laboratories and in controlled clinical settings. Several but not all of the studies suggest that higher blood pressure levels are associated with the tendency not to recall or report occurrences identified as racist and discriminatory. Investigators have reported that physiological arousal is associated with laboratory analogues of ethnic discrimination and mistreatment. Evidence from survey and laboratory studies suggests that personality variables and cultural orientation moderate the impact of racial discrimination. The neural pathways that mediate these physiological reactions are not known. The evidence supports the notion that direct encounters with discriminatory events contribute to negative health outcomes. PMID- 12554579 TI - Violence and health: the United States in a global perspective. AB - Violence is a public health problem that can be understood and changed. Research over the past 2 decades has demonstrated that violence can be prevented and that, in some cases, prevention programs are more cost-effective than other policy options such as incarceration. The United States has much to contribute to-and stands to gain much from-global efforts to prevent violence. A new World Health Organization initiative presents an opportunity for the United States to work with other nations to find cost-effective ways of preventing violence and reducing its enormous costs. PMID- 12554578 TI - Paved with good intentions: do public health and human service providers contribute to racial/ethnic disparities in health? AB - There is extensive evidence of racial/ethnic disparities in receipt of health care. The potential contribution of provider behavior to such disparities has remained largely unexplored. Do health and human service providers behave in ways that contribute to systematic inequities in care and outcomes? If so, why does this occur? The authors build on existing evidence to provide an integrated, coherent, and sound approach to research on providers' contributions to racial/ethnic disparities. They review the evidence regarding provider contributions to disparities in outcomes and describe a causal model representing an integrated set of hypothesized mechanisms through which health care providers' behaviors may contribute to these disparities. PMID- 12554580 TI - Prejudice as stress: conceptual and measurement problems. AB - In the field of social sciences, there has been a renewed interest in studying prejudice and discrimination as stressors and assessing their impact on various health outcomes. This raises a need for theoretically based and psychometrically sound measures of prejudice. As researchers approach this task, there are several conceptual issues that need to be addressed. The author describes 3 such issues related to (1) individual versus structural measures of the impact of prejudice, (2) objective versus subjective assessments of stress, and (3) measures of major events versus everyday discrimination. How researchers approach the problem of measurement depends on the specific study aims, but they must consider these conceptual issues and understand the advantages and limitations of various approaches to the study of prejudice as stress. PMID- 12554582 TI - A well baby clinic in Indianapolis. PMID- 12554581 TI - Smoking and ill health: does lay epidemiology explain the failure of smoking cessation programs among deprived populations? AB - The resistance of disadvantaged groups to anti-smoking advice is remarkable. In relation to the study of differing cultures, there is a long-standing academic tradition assuming that behavior that may otherwise be difficult to understand is indeed rational within particular cultural contexts. Persistent smoking among the most deprived members of society may represent a rational response to their life chances informed by a lay epidemiology. Health promotion initiatives designed to reduce smoking among members of these groups may continue to fail unless the general health and life chances of such individuals are first improved. PMID- 12554583 TI - The health and physique of the Negro American. 1906. PMID- 12554584 TI - William Edward Burghardt DuBois: historian, social critic, activist. PMID- 12554585 TI - The structuring of ethnic inequalities in health: economic position, racial discrimination, and racism. AB - Differences in health across ethnic groups have been documented in the United States and the United Kingdom. The extent to which socioeconomic inequalities underlie such differences remains contested, with many instead focusing on cultural or genetic explanations. In both the United States and the United Kingdom, data limitations have greatly hampered investigations of ethnic inequalities in health. Perhaps foremost of these is the inadequate measurement of ethnicity, but also important is the lack of good data on socioeconomic position, particularly data that address life-course issues. Other elements of social disadvantage, particularly experiences of racism, are also neglected. The author reviews existing evidence and presents new evidence to suggest that social and economic inequalities, underpinned by racism, are fundamental causes of ethnic inequalities in health. PMID- 12554587 TI - Happiness and children's health: an investigation of art, entertainment, and recreation. PMID- 12554586 TI - The health status of newly arrived refugee children in Miami-Dade County, Florida. PMID- 12554588 TI - Identifying risk factors for rubella susceptibility in a population at risk in the United States. PMID- 12554589 TI - Cancer-related health disparities in women. AB - OBJECTIVES: This article synthesizes information about cancer in 9 populations of minority women: Mexican American, Puerto Rican, Cuban American, African American, Asian American, Native Hawaiian, American Samoan, American Indian, and Alaska Native. METHODS: Cancer registry data, social indicators, government sources, and published articles were searched for information on the background and cancer experience of these 9 racial/ethnic groups. RESULTS: Approximately 35 million women in these racial/ethnic groups live in the United States, and their numbers are increasing rapidly. Since 1992, incidence rates for major cancer sites have slowed or decreased among these groups, but declines in mortality have not occurred or have been smaller than for Whites. Gaps in early detection have narrowed, but minority women still lag behind Whites. Smoking and obesity remain common in these populations. CONCLUSIONS: More culturally appropriate interventions and research are needed, and these efforts must involve the community and raise the quality of health services. PMID- 12554590 TI - Onset of natural menopause in African American women. AB - OBJECTIVES: This study assessed predictors of the onset of natural menopause in African American women. METHODS: We used mailed questionnaires to collect data at baseline in 1995 and during follow-up from Black Women's Health Study participants. Cox proportional hazards regression was used to assess potential predictors-including experiences of racism-of the onset of natural menopause among 17 070 women aged 35 to 55 years and premenopausal in 1995. RESULTS: The hazard ratio (HR) was 1.43 for current smokers (95% confidence interval [CI] = 1.24, 1.66) and 1.21 (95% CI = 1.06, 1.38) for ex-smokers and significantly less for obese women and oral contraceptive users. Hazard ratios for most questions about racism were elevated by 10% to 30% but were not statistically significant. CONCLUSIONS: Earlier onset of natural menopause among African American women is strongly associated with smoking and inversely associated with body mass index and oral contraceptive use. PMID- 12554591 TI - Hysterectomy prevalence by Hispanic ethnicity: evidence from a national survey. AB - OBJECTIVES: We investigated hysterectomy prevalence among Hispanic women. METHODS: We obtained data from 4684 Hispanic women and 20 604 non-Hispanic White women from the 1998-1999 National Health Interview Survey. We calculated nationally representative odds ratios of previous hysterectomy, controlling for confounders. RESULTS: Compared with non-Hispanic White women, the odds ratio for hysterectomy was 0.36 (95% confidence interval [CI] = 0.30, 0.44) for Hispanic women with no high school diploma, 0.57 (95% CI = 0.44, 0.74) for high school graduates, and 0.67 (95% CI = 0.42, 0.87) for college attenders. Country of origin had little influence on hysterectomy prevalence. Hysterectomy was positively associated with acculturation. CONCLUSIONS: Hispanic women undergo fewer hysterectomies than do non-Hispanic White women. The reasons for this, as well as information on ethnicity-specific appropriateness of hysterectomy, should be explored. PMID- 12554592 TI - A synthesis of perceptions about physical activity among older African American and American Indian women. AB - OBJECTIVES: In this review of qualitative data from the Cross-Cultural Activity Participation Study (CAPS), we synthesize the major findings of studies designed to identify minority women's perceptions of physical activity. METHODS: We interviewed 30 African American and 26 American Indian women with constant comparison techniques. We analyzed the data with a coding system developed from the data. RESULTS: The women led active, busy lives. Most perceived physical activity as being good for them, identified constraints to time and space for physical activity, and wanted social support for physical activity. Sociocultural issues also were related to physical activity. CONCLUSIONS: Both personal and cultural values influenced the women's physical activity behaviors. PMID- 12554593 TI - Relation of dietary quality, physical activity, and smoking habits to 10-year changes in health status in older Europeans in the SENECA study. AB - OBJECTIVES: This study investigated the effect of healthy lifestyle behaviors on self-rated health and self-care ability over a 10-year follow-up period in older persons in the SENECA study. METHODS: Health status and lifestyle behaviors were examined in 1988/1989, 1993, and 1999 in 216 men and 264 women, born between 1913 and 1918, from 7 European countries. RESULTS: Self-rated health and self-care ability declined in men and women with healthy and unhealthy lifestyle habits over the 10-year follow-up period. Inactive and smoking persons had an increased risk for a decline in health status as compared with active and nonsmoking people. No effect of a healthy, Mediterranean-like diet on the deterioration in health status was observed. CONCLUSIONS: Being physically active and nonsmoking delayed deterioration in health status in older participants aged 70 to 75 years in the SENECA study. PMID- 12554594 TI - Racial/ethnic differences in adult vaccination among individuals with diabetes. AB - OBJECTIVES: This study examined whether differences in access to health care, health coverage, and socioeconomic status (SES) explained racial differences in influenza and pneumococcal vaccination rates in individuals with diabetes. METHODS: We analyzed data on 1906 individuals from the 1998 National Health Interview Survey. We used multiple logistic regression to adjust for race/ethnicity, age, access to care, health insurance, and SES, and used SUDAAN for statistical analyses to yield national estimates. RESULTS: Whites had higher vaccination rates than did African Americans or Hispanics. After adjustment for covariates, race/ethnicity predicted receipt of both vaccines independent of age, access to care, health care coverage, and SES. CONCLUSIONS: Racial disparity in vaccination rates for adults with diabetes is independent of access to care, health care coverage, and SES. PMID- 12554595 TI - Does assistive technology substitute for personal assistance among the disabled elderly? AB - OBJECTIVES: This study examined whether use of equipment (technological assistance) to cope with disability was associated with use of fewer hours of help from another person (personal assistance). METHODS: In a cross-sectional study of 2368 community dwellers older than 65 years with 1 or more limitations in basic activities of daily living (ADLs) from the 1994 National Long Term Care Survey, the relation between technological assistance and personal assistance was examined. RESULTS: Among people with ADL limitations, multivariate models showed a strong and consistent relation between technological assistance and personal assistance, whereby use of equipment was associated with fewer hours of help. CONCLUSIONS: Among people with disability, use of assistive technology was associated with use of fewer hours of personal assistance. PMID- 12554596 TI - Greater replication and differentiation of preadipocytes in inherited corticosteroid-binding globulin deficiency. AB - Glucocorticoids are pivotal for adipose tissue development. Rodent studies suggest that corticosteroid-binding globulin (CBG) modulates glucocorticoid action in adipose tissue. In humans, both genetic CBG deficiency and suppressed CBG concentrations in hyperinsulinemic states are associated with obesity. We hypothesized that CBG deficiency in humans modulates the response of human preadipocytes to glucocorticoids, predisposing them to obesity. We compared normal preadipocytes with subcultured preadipocytes from an individual with the first ever described complete deficiency of CBG due to a homozygous null mutation. CBG-negative preadipocytes proliferated more rapidly and showed greater peroxisome proliferator-activated receptor-gamma-mediated differentiation than normal preadipocytes. CBG was not expressed in normal human preadipocytes. Glucocorticoid receptor number and binding characteristics and 11beta hydroxysteroid dehydrogenase activity were similar for CBG-negative and normal preadipocytes. We propose that the increased proliferation and enhanced differentiation of CBG-negative preadipocytes may promote adipose tissue deposition and explain the obesity seen in individuals with genetic CBG deficiency. Furthermore, these observations may be relevant to obesity occurring with suppressed CBG concentrations associated with hyperinsulinemia. PMID- 12554597 TI - Visceral fat and liver fat are independent predictors of metabolic risk factors in men. AB - We examined the independent associations among abdominal adipose tissue (AT), liver fat, cardiorespiratory fitness (CRF), and lipid variables in 161 Caucasian men who had a wide variation in adiposity. We measured AT and liver fat by computed tomography and CRF by a maximal exercise test on a treadmill. Visceral AT remained a significant (P or= 0.05) correlate of any lipid variable after control for visceral AT and CRF. Furthermore, subdivision of subcutaneous AT into deep and superficial depots did not alter these observations. Visceral AT was the strongest correlate of liver fat and remained so after control for abdominal subcutaneous AT, CRF, and alcohol consumption (r = -0.34, P < 0.01). In contrast, abdominal subcutaneous AT and CRF were not significant (P > 0.10) correlates of liver fat after control for visceral AT. Visceral AT remained a significant (P < 0.01) correlate of TG, HDL C, and TC/HDL-C independent of liver fat. However, liver fat was also a significant correlate (P 10-fold, respectively, in renal tubule epithelial cells subjected to ATP depletion and metabolic poisoning (an in vitro model of kidney IRI). Our results suggest that SSAT is likely a new marker of tubular cell injury that distinguishes acute prerenal from intrarenal failure. PMID- 12554637 TI - From Alzheimer to Huntington: why is a structural understanding so difficult? AB - An increasing family of neurodegenerative disorders such as Alzheimer's, Parkinson's and Huntington's diseases, prion encephalopathies and cystic fibrosis is associated with aggregation of misfolded polypeptide chains which are toxic to the cell. Knowledge of the three-dimensional structure of the proteins implicated is essential for understanding why and how endogenous proteins may adopt a non native fold. Yet, structural work has been hampered by the difficulty of handling proteins insoluble or prone to aggregation, and at the same time that is why it is interesting to study these molecules. In this review, we compare the structural knowledge accumulated for two paradigmatic misfolding disorders, Alzheimer's disease (AD) and the family of poly-glutamine diseases (poly-Q) and discuss some of the hypotheses suggested for explaining aggregate formation. While a common mechanism between these pathologies remains to be proven, a direct comparison may help in designing new strategies for approaching their study. PMID- 12554638 TI - Two distinct myosin light chain structures are induced by specific variations within the bound IQ motifs-functional implications. AB - IQ motifs are widespread in nature. Mlc1p is a calmodulin-like myosin light chain that binds to IQ motifs of a class V myosin, Myo2p, and an IQGAP-related protein, Iqg1p, playing a role in polarized growth and cytokinesis in Saccharomyces cerevisiae. The crystal structures of Mlc1p bound to IQ2 and IQ4 of Myo2p differ dramatically. When bound to IQ2, Mlc1p adopts a compact conformation in which both the N- and C-lobes interact with the IQ motif. However, in the complex with IQ4, the N-lobe no longer interacts with the IQ motif, resulting in an extended conformation of Mlc1p. The two light chain structures relate to two distinct subfamilies of IQ motifs, one of which does not interact with the N-lobes of calmodulin-like light chains. The correlation between light chain structure and IQ sequence is demonstrated further by sedimentation velocity analysis of complexes of Mlc1p with IQ motifs from Myo2p and Iqg1p. The resulting 'free' N lobes of myosin light chains in the extended conformation could mediate the formation of ternary complexes during protein localization and/or partner recruitment. PMID- 12554639 TI - SOCS3: an essential regulator of LIF receptor signaling in trophoblast giant cell differentiation. AB - Suppressor of cytokine signaling 3 (SOCS3) binds cytokine receptors and thereby suppresses cytokine signaling. Deletion of SOCS3 causes an embryonic lethality that is rescued by a tetraploid rescue approach, demonstrating an essential role in placental development and a non-essential role in embryo development. Rescued SOCS3-deficient mice show a perinatal lethality with cardiac hypertrophy. SOCS3 deficient placentas have reduced spongiotrophoblasts and increased trophoblast secondary giant cells. Enforced expression of SOCS3 in a trophoblast stem cell line (Rcho-1) suppresses giant cell differentiation. Conversely, SOCS3-deficient trophoblast stem cells differentiate more readily to giant cells in culture, demonstrating that SOCS3 negatively regulates trophoblast giant cell differentiation. Leukemia inhibitory factor (LIF) promotes giant cell differentiation in vitro, and LIF receptor (LIFR) deficiency results in loss of giant cell differentiation in vivo. Finally, LIFR deficiency rescues the SOCS3 deficient placental defect and embryonic lethality. The results establish SOCS3 as an essential regulator of LIFR signaling in trophoblast differentiation. PMID- 12554640 TI - Restriction of multiple divergent retroviruses by Lv1 and Ref1. AB - The mouse gene Fv1 encodes a saturable restriction factor that selectively blocks infection by N-tropic or B-tropic murine leukemia virus (MLV) strains. Despite the absence of an Fv1 gene, a similar activity is present in humans that blocks N MLV infection (Ref1). Moreover, some non-human primate cell lines express a potentially related inhibitor of HIV-1 and/or SIVmac infection (Lv1). Here, we examine the spectrum of retrovirus-restricting activities expressed by human and African green monkey cell lines. Human cells restrict N-MLV and equine infectious anemia virus (EIAV), but not HIV-1, HIV-2, SIVmac or SIVagm, whilst AGM cells restrict N-MLV, EIAV, HIV-1, HIV-2 and SIVmac. Remarkably, in each example examined, restriction of infection by a given retrovirus can be abrogated at least partially by saturation with another retrovirus, provided that it is also restricted but regardless of whether it is closely related. These data suggest that restriction factors in human and non-human primate cells are able to recognize and block infection by multiple, widely divergent retroviruses and that the factors themselves may be related. PMID- 12554641 TI - C-terminal domains implicated in the functional surface expression of potassium channels. AB - A short C-terminal domain is required for correct tetrameric assembly in some potassium channels. Here, we show that this domain forms a coiled coil that determines not only the stability but also the selectivity of the multimerization. Synthetic peptides comprising the sequence of this domain in Eag1 and other channels are able to form highly stable tetrameric coiled coils and display selective heteromultimeric interactions. We show that loss of function caused by disruption of this domain in Herg1 can be rescued by introducing the equivalent domain from Eag1, and that this chimeric protein can form heteromultimers with Eag1 while wild-type Erg1 cannot. Additionally, a short endoplasmic reticulum retention sequence closely preceding the coiled coil plays a crucial role for surface expression. Both domains appear to co-operate to form fully functional channels on the cell surface and are a frequent finding in ion channels. Many pathological phenotypes may be attributed to mutations affecting one or both domains. PMID- 12554642 TI - Scrapie-like prion protein accumulates in aggresomes of cyclosporin A-treated cells. AB - Prion diseases are infectious, sporadic and inherited fatal neurodegenerations that are propagated by an abnormal refolding of the cellular prion protein PrP(C). Which chaperones assist the normal folding of PrP(C) is unknown. The linkage of familial Gerstmann- Straussler-Scheinker (GSS) syndrome with proline substitutions in PrP raised the prospect that peptidylprolyl cis-trans isomerases (PPIases) may play a role in normal PrP metabolism. Here we used cyclo sporin A (CsA), an immunosuppressant, to inhibit the cyclophilin family of PPIases in cultured cells. CsA-treated cells accumulated proteasome-resistant, 'prion-like' PrP species, which deposited in long-lived aggresomes. PrP aggresomes also formed with disease-linked proline mutants when proteasomes were inhibited. These results suggest mechanisms whereby abnormally folded cytosolic PrP may in some cases participate in the development of spontaneous and inherited prion diseases. PMID- 12554643 TI - H+/ATP ratio of proton transport-coupled ATP synthesis and hydrolysis catalysed by CF0F1-liposomes. AB - The H(+)/ATP ratio and the standard Gibbs free energy of ATP synthesis were determined with a new method using a chemiosmotic model system. The purified H(+) translocating ATP synthase from chloroplasts was reconstituted into phosphatidylcholine/phosphatidic acid liposomes. During reconstitution, the internal phase was equilibrated with the reconstitution medium, and thereby the pH of the internal liposomal phase, pH(in), could be measured with a conventional glass electrode. The rates of ATP synthesis and hydrolysis were measured with the luciferin/luciferase assay after an acid-base transition at different [ATP]/([ADP][P(i)]) ratios as a function of deltapH, analysing the range from the ATP synthesis to the ATP hydrolysis direction and the deltapH at equilibrium, deltapH (eq) (zero net rate), was determined. The analysis of the [ATP]/([ADP][P(i)]) ratio as a function of deltapH (eq) and of the transmembrane electrochemical potential difference, delta micro approximately (H)(+) (eq), resulted in H(+)/ATP ratios of 3.9 +/- 0.2 at pH 8.45 and 4.0 +/- 0.3 at pH 8.05. The standard Gibbs free energies of ATP synthesis were determined to be 37 +/- 2 kJ/mol at pH 8.45 and 36 +/- 3 kJ/mol at pH 8.05. PMID- 12554644 TI - SufC: an unorthodox cytoplasmic ABC/ATPase required for [Fe-S] biogenesis under oxidative stress. AB - Proteins containing [Fe-S] clusters perform essential functions in all domains of life. Previously, we identified the sufABCDSE operon as being necessary for virulence of the plant pathogen Erwinia chrysanthemi. In addition, we collected preliminary evidence that the sufABCDSE operon might be involved in the assembly of [Fe-S] clusters. Of particular interest are the sufB, sufC and sufD genes, which are conserved among Eubacteria, Archaea, plants and parasites. The present study establishes SufC as an unorthodox ATPase of the ABC superfamily that is located in the cytosol, wherein it interacts with both SufB and SufD. Moreover, under oxidative stress conditions, SufC was found to be necessary for the activity of enzymes containing oxygen-labile [Fe-S] clusters, but dispensable for glutamate synthase, which contains an oxidatively stable [Fe-S] cluster. Lastly, we have shown SufBCD to be essential for iron acquisition via chrysobactin, a siderophore of major importance in virulence. We discuss a model wherein the SufBCD proteins contribute to bacterial pathogenicity via their role in the assembly of [Fe-S] clusters under oxidative stress and iron limitation. PMID- 12554645 TI - Yeast Cdk1 translocates to the plus end of cytoplasmic microtubules to regulate bud cortex interactions. AB - The budding yeast spindle aligns along the mother- bud axis through interactions between cytoplasmic microtubules (CMs) and the cell cortex. Kar9, in complex with the EB1-related protein Bim1, mediates contacts of CMs with the cortex of the daughter cell, the bud. Here we established a novel series of events that target Kar9 to the bud cortex. First, Kar9 binds to spindle pole bodies (SPBs) in G(1) of the cell cycle. Secondly, in G(1)/S the yeast Cdk1, Cdc28, associates with SPBs and phosphorylates Kar9. Thirdly, Kar9 and Cdc28 then move from the SPB to the plus end of CMs directed towards the bud. This movement is dependent upon the microtubule motor protein Kip2. Cdc28 activity is required to concentrate Kar9 at the plus end of CMs and hence to establish contacts with the bud cortex. The Cdc28-regulated localization of Kar9 is therefore an integral part of the program that aligns spindles. PMID- 12554646 TI - A conserved tyrosine in the neck of a fungal kinesin regulates the catalytic motor core. AB - The neck domain of fungal conventional kinesins displays characteristic properties which are reflected in a specific sequence pattern. The exchange of the strictly conserved Tyr 362, not present in animals, into Lys, Cys or Phe leads to a failure to dimerize. The destabilizing effect is confirmed by a lower coiled-coil propensity of mutant peptides. Whereas the Phe substitution has only a structural effect, the Lys and Cys replacements lead to dramatic kinetic changes. The steady state ATPase is 4- to 7-fold accelerated, which may be due to a faster microtubule-stimulated ADP release rate. These data suggest that an inhibitory effect of the fungal neck domain on the motor core is mediated by direct interaction of the aromatic ring of Tyr 362 with the head, whereas the OH group is essential for dimerization. This is the first demonstration of a direct influence of the kinesin neck region in regulation of the catalytic activity. PMID- 12554647 TI - Regulation of light-dependent Gqalpha translocation and morphological changes in fly photoreceptors. AB - Heterotrimeric G-proteins relay signals between membrane-bound receptors and downstream effectors. Little is known, however, about the regulation of Galpha subunit localization within the natural endogenous environment of a specialized signaling cell. Here we show, using live Drosophila flies, that light causes massive and reversible translocation of the visual Gqalpha to the cytosol, associated with marked architectural changes in the signaling compartment. Molecular genetic dissection together with detailed kinetic analysis enabled us to characterize the translocation cycle and to unravel how signaling molecules that interact with Gqalpha affect these processes. Epistatic analysis showed that Gqalpha is necessary but not sufficient to bring about the morphological changes in the signaling organelle. Furthermore, mutant analysis indicated that Gqbeta is essential for targeting of Gqalpha to the membrane and suggested that Gqbeta is also needed for efficient activation of Gqalpha by rhodopsin. Our results support the 'two-signal model' hypothesis for membrane targeting in a living organism and characterize the regulation of both the activity-dependent Gq localization and the cellular architectural changes in Drosophila photoreceptors. PMID- 12554648 TI - PDE5 is converted to an activated state upon cGMP binding to the GAF A domain. AB - cGMP-specific, cGMP-binding phosphodiesterase (PDE5) regulates such physiological processes as smooth muscle relaxation and neuronal survival. PDE5 contains two N terminal domains (GAF A and GAF B), but the functional roles of these domains have not been determined. Here we show that recombinant PDE5 is activated directly upon cGMP binding to the GAF A domain, and this effect does not require PDE5 phosphorylation. PDE5 exhibited time- and concentration-dependent reversible activation in response to cGMP, with the highest activation (9- to 11-fold) observed at low substrate concentrations (0.1 micro M cGMP). A monoclonal antibody directed against GAF A blocked cGMP binding, prevented PDE5 activation and decreased basal activity, revealing that PDE5 in its non-activated state has low intrinsic catalytic activity. Activated PDE5 showed higher sensitivity towards sildenafil than non-activated PDE5. The stimulatory effect of cGMP binding on the catalytic activity of PDE5 suggests that this mechanism of enzyme activation may be common among other GAF domain-containing proteins. The data also suggest that development of agonists and antagonists of PDE5 activity based on binding to this site might be possible. PMID- 12554649 TI - Identification of YB-1 as a regulator of PTP1B expression: implications for regulation of insulin and cytokine signaling. AB - Changes in expression of PTP1B, the prototypic protein tyrosine phosphatase, have been associated with various human diseases; however, the mechanisms by which PTP1B expression is regulated have not been defined. We have identified an enhancer sequence within the PTP1B promoter which serves as a binding site for the transcription factor Y box-binding protein-1 (YB-1). Overexpression of YB-1 resulted in increased levels of PTP1B. Furthermore, depletion of YB-1 protein, by expression of a specific antisense construct, led to an approximately 70% decrease in expression of PTP1B, but no change in the level of its closest relative, TC-PTP. Expression of antisense YB-1 resulted in increased sensitivity to insulin and enhanced signaling through the cytokine receptor gp130, which was suppressed by re-expression of PTP1B. Finally, we observed a correlation between the expression of PTP1B and that of YB-1 in cancer cell lines and an animal model of type II diabetes. Our data reveal an important role for YB-1 as a regulator of PTP1B expression, and further highlight PTP1B as a critical regulator of insulin- and cytokine-mediated signal transduction. PMID- 12554650 TI - Structural basis for recruitment of glycogen synthase kinase 3beta to the axin APC scaffold complex. AB - Glycogen synthase kinase 3beta (GSK3beta) is a serine/threonine kinase involved in insulin, growth factor and Wnt signalling. In Wnt signalling, GSK3beta is recruited to a multiprotein complex via interaction with axin, where it hyperphosphorylates beta-catenin, marking it for ubiquitylation and destruction. We have now determined the crystal structure of GSK3beta in complex with a minimal GSK3beta-binding segment of axin, at 2.4 A resolution. The structure confirms the co-localization of the binding sites for axin and FRAT in the C terminal domain of GSK3beta, but reveals significant differences in the interactions made by axin and FRAT, mediated by conformational plasticity of the 285-299 loop in GSK3beta. Detailed comparison of the axin and FRAT GSK3beta complexes allows the generation of highly specific mutations, which abrogate binding of one or the other. Quantitative analysis suggests that the interaction of GSK3beta with the axin scaffold enhances phosphorylation of beta-catenin by >20 000-fold. PMID- 12554651 TI - Structural basis of adhesion-molecule recognition by ERM proteins revealed by the crystal structure of the radixin-ICAM-2 complex. AB - ERM (ezrin/radixin/moesin) proteins recognize the cytoplasmic domains of adhesion molecules in the formation of the membrane-associated cytoskeleton. Here we report the crystal structure of the radixin FERM (4.1 and ERM) domain complexed with the ICAM-2 cytoplasmic peptide. The non-polar region of the ICAM-2 peptide contains the RxxTYxVxxA sequence motif to form a beta-strand followed by a short 3(10)-helix. It binds the groove of the phosphotyrosine-binding (PTB)-like subdomain C mediated by a beta-beta association and several side-chain interactions. The binding mode of the ICAM-2 peptide to the FERM domain is distinct from that of the NPxY motif-containing peptide binding to the canonical PTB domain. Mutation analyses based on the crystal structure reveal the determinant elements of recognition and provide the first insights into the physical link between adhesion molecules and ERM proteins. PMID- 12554652 TI - The Helicobacter pylori CagA protein induces cortactin dephosphorylation and actin rearrangement by c-Src inactivation. AB - The gastric pathogen Helicobacter pylori translocates the CagA protein into epithelial cells by a type IV secretion process. Translocated CagA is tyrosine phosphorylated (CagA(P-Tyr)) on specific EPIYA sequence repeats by Src family tyrosine kinases. Phos phorylation of CagA induces the dephosphorylation of as yet unidentified cellular proteins, rearrangements of the host cell actin cytoskeleton and cell scattering. We show here that CagA(P-Tyr) inhibits the catalytic activity of c-Src in vivo and in vitro. c-Src inactivation leads to tyrosine dephosphorylation of the actin binding protein cortactin. Concomitantly, cortactin is specifically redistributed to actin-rich cellular protrusions. c-Src inactivation and cortactin dephosphorylation are required for rearrangements of the actin cytoskeleton. Moreover, CagA(P-Tyr)-mediated c-Src inhibition downregulates further CagA phosphorylation through a negative feedback loop. This is the first report of a bacterial virulence factor that inhibits signalling of a eukaryotic tyrosine kinase and on a role of c-Src inactivation in host cell cytoskeletal rearrangements. PMID- 12554653 TI - Mapping of the laminin-binding site of the N-terminal agrin domain (NtA). AB - Agrin is a key organizer of acetylcholine receptor (AChR) clustering at the neuromuscular junction. The binding of agrin to laminin is required for its localization to synaptic basal lamina and other basement membranes. The high affinity interaction with the coiled-coil domain of laminin is mediated by the N terminal domain of agrin. We have adopted a structurally guided site-directed mutagenesis approach to map the laminin-binding site of NtA. Mutations of L117 and V124 in the C-terminal helix 3 showed that they are crucial for binding. Both residues are located in helix 3 and face the groove between the beta-barrel and the C-terminal helical segment of NtA. Remarkably, the distance between both residues matches a heptad repeat distance of two aliphatic residues which are solvent exposed in the coiled-coil domain of laminin. A lower but significant contribution originates from R43 and a charged cluster (E23, E24 and R40) at the open face of the beta-barrel structure. We propose that surface-exposed, conserved residues of the laminin gamma1 chain interact with NtA via hydrophobic and ionic interactions. PMID- 12554654 TI - The tyrosine kinase Tyk2 controls IFNAR1 cell surface expression. AB - The four mammalian Jak tyrosine kinases are non-covalently associated with cell surface receptors binding helical bundled cytokines. In the type I interferon receptor, Tyk2 associates with the IFNAR1 receptor subunit and positively influences ligand binding to the receptor complex. Here, we report that Tyk2 is essential for stable cell surface expression of IFNAR1. In the absence of Tyk2, mature IFNAR1 is weakly expressed on the cell surface. Rather, it is localized into a perinuclear endosomal compartment which overlaps with that of recycling transferrin receptors and with early endosomal antigen-1 (EEA1) positive vesicles. Conversely, co-expressed Tyk2 greatly enhances surface IFNAR1 expression. Importantly, we demonstrate that Tyk2 slows down IFNAR1 degradation and that this is due, at least in part, to inhibition of IFNAR1 endocytosis. In addition, Tyk2 induces plasma membrane relocalization of the R2 subunit of the interleukin-10 receptor. These results reveal a novel function of a Jak protein on internalization of a correctly processed cytokine receptor. This function is distinct from the previously reported effect of other Jak proteins on receptor exit from the endoplasmic reticulum. PMID- 12554655 TI - Retromer and the sorting nexins Snx4/41/42 mediate distinct retrieval pathways from yeast endosomes. AB - The endocytic pathway in yeast leads to the vacuole, but resident proteins of the late Golgi, and some endocytosed proteins such as the exocytic SNARE Snc1p, are retrieved specifically to the Golgi. Retrieval can occur from both a late pre vacuolar compartment and early or 'post-Golgi' endosomes. We show that the endosomal SNARE Pep12p, and a mutant version that reaches the cell surface and is endocytosed, are retrieved from pre-vacuolar endosomes. As with Golgi proteins, this requires the sorting nexin Grd19p and components of the retromer coat, supporting the view that endosomal and Golgi residents both cycle continuously between the exocytic and endocytic pathways. In contrast, retrieval of Snc1p from post-Golgi endosomes requires the sorting nexin Snx4p, to which Snc1p can be cross-linked. Snx4p binds to Snx41p/ydr425w and to Snx42p/ydl113c, both of which are also required for efficient Snc1p sorting. Our findings suggest a general role for yeast sorting nexins in protein retrieval, rather than degradation, and indicate that different sorting nexins operate in different classes of endosomes. PMID- 12554656 TI - Regulated delivery of AMPA receptor subunits to the presynaptic membrane. AB - In recent years, a role for AMPA receptors as modulators of presynaptic functions has emerged. We have investigated the presence of AMPA receptor subunits and the possible dynamic control of their surface exposure at the presynaptic membrane. We demonstrate that the AMPA receptor subunits GluR1 and GluR2 are expressed and organized in functional receptors in axonal growth cones of hippocampal neurons. AMPA receptors are actively internalized upon activation and recruited to the surface upon depolarization. Pretreatment of cultures with botulinum toxin E or tetanus toxin prevents the receptor insertion into the plasma membrane, whereas treatment with alpha-latrotoxin enhances the surface exposure of GluR2, both in growth cones of cultured neurons and in brain synaptosomes. Purification of small synaptic vesicles through controlled-pore glass chromatography, revealed that both GluR2 and GluR1, but not the GluR2 interacting protein GRIP, copurify with synaptic vesicles. These data indicate that, at steady state, a major pool of AMPA receptor subunits reside in synaptic vesicle membranes and can be recruited to the presynaptic membrane as functional receptors in response to depolarization. PMID- 12554657 TI - Loss of myosin VI reduces secretion and the size of the Golgi in fibroblasts from Snell's waltzer mice. AB - Golgi morphology and function are dependent on an intact microtubule and actin cytoskeleton. Myosin VI, an unusual actin-based motor protein moving towards the minus ends of actin filaments, has been localized to the Golgi complex at the light and electron microscopic level. Myosin VI is present in purified Golgi membranes as a peripheral membrane protein, targeted by its globular tail domain. To investigate the function of myosin VI at the Golgi complex, immortal fibroblastic cell lines of Snell's waltzer mice lacking myosin VI were established. In these cell lines, where myosin VI is absent, the Golgi complex is reduced in size by approximately 40% compared with wild-type cells. Furthermore, protein secretion of a reporter protein from Snell's waltzer cells is also reduced by 40% compared with wild-type cells. Rescue experiments showed that fully functional myosin VI was able to restore Golgi complex morphology and protein secretion in Snell's waltzer cells to the same level as that observed in wild-type cells. PMID- 12554658 TI - Nitric oxide-induced suspended animation promotes survival during hypoxia. AB - Oxygen plays a key role in energy metabolism. However, there are organisms that survive severe shortfalls in oxygen. Drosophila embryos rapidly arrest development upon severe hypoxia and recover upon restoration of oxygen, even days later. Stabilization of the normally unstable engrailed RNA and protein preserved the localized striped pattern of this embryonic patterning gene during 3 days in hypoxia. Severe hypoxia blocked expression of a heat-shock-inducible lacZ transgene. Cyanide, a metabolic poison, did not immediately block gene expression or turnover, arguing against a passive response to energy limitation. In contrast, nitric oxide, a putative hypoxia signal, induced a reversible arrest of development, gene expression and turnover. Reciprocally, a nitric oxide scavenger allowed continued gene expression and turnover during hypoxia, but it reduced hypoxia tolerance. We suggest that hypoxia-induced stasis preserves the status quo of embryonic processes and promotes survival. Our data implicate nitric oxide as a mediator of this response and provide a system in which to investigate its action. PMID- 12554659 TI - Histone H1 enhances synergistic activation of the MMTV promoter in chromatin. AB - Minichromosomes assembled on the mouse mammary tumor virus (MMTV) promoter in vitro exhibit positioned nucleosomes, one of which covers the binding sites for progesterone receptor (PR) and nuclear factor 1 (NF1). Incorporation of histone H1 into MMTV minichromosomes improves the stability of this nucleosome and decreases basal transcription from the MMTV promoter, as well as its response to either PR or NF1. However, histone H1-containing minichromosomes display better PR binding and support a more efficient synergism between PR and NF1, leading to enhanced transcription initiation. A mutant MMTV promoter lacking positioned nucleosomes does not display enhanced transcriptional synergism in the presence of H1. Binding of PR leads to phosphorylation of H1, which leaves the promoter upon transcription initiation. Thus, H1 assumes a complex and dynamic role in the regulation of the MMTV promoter. PMID- 12554660 TI - Chromatin exposes intrinsic differences in the transcriptional activities of estrogen receptors alpha and beta. AB - The biological actions of estrogens are mediated via two distinct intranuclear estrogen receptor (ER) proteins, ERalpha and ERbeta. We have used an in vitro chromatin assembly and transcription system to compare the transcriptional activities of the two ERs in the context of chromatin, the physiological template for transcription by RNA polymerase II. We find that under conditions where many biochemical activities of the receptors are similar (e.g. ligand binding, chromatin binding, chromatin remodeling and co-activator recruitment), liganded ERalpha is a much more potent transcriptional activator than ERbeta with chromatin templates, but not with naked DNA. This difference is attributable to the N-terminal A/B region of ERalpha, which contains a transferable activation function that facilitates transcription specifically with chromatin templates. Interestingly, chromatin selectively restricts ligand-dependent transcriptional activation by ERbeta under some conditions (e.g. with a closed chromatin architecture), while allowing it under other conditions (e.g. with an open chromatin architecture). Collectively, our results define an important role for chromatin in determining signaling outcomes mediated by distinct subtypes of signal-transducing transcriptional activator proteins. PMID- 12554661 TI - Molecular evidence for a positive role of Spt4 in transcription elongation. AB - We have previously shown that yeast mutants of the THO complex have a defect in gene expression, observed as an impairment of lacZ transcription. Here we analyze the ability of mutants of different transcription elongation factors to transcribe lacZ. We found that spt4Delta, like THO mutants, impaired transcription of lacZ and of long and GC-rich DNA sequences fused to the GAL1 promoter. Using a newly developed in vitro transcription elongation assay, we show that Spt4 is required in elongation. There is a functional interaction between Spt4 and THO, detected by the lethality or strong gene expression defect and hyper-recombination phenotypes of double mutants in the W303 genetic background. Our results indicate that Spt4-Spt5 has a positive role in transcription elongation and suggest that Spt4-Spt5 and THO act at different steps during mRNA biogenesis. PMID- 12554662 TI - Plant dicistronic tRNA-snoRNA genes: a new mode of expression of the small nucleolar RNAs processed by RNase Z. AB - Small nucleolar RNAs (snoRNAs) guiding modifications of ribosomal RNAs and other RNAs display diverse modes of gene organization and expression depending on the eukaryotic system: in animals most are intron encoded, in yeast many are monocistronic genes and in plants most are polycistronic (independent or intronic) genes. Here we report an unprecedented organization: plant dicistronic tRNA-snoRNA genes. In Arabidopsis thaliana we identified a gene family encoding 12 novel box C/D snoRNAs (snoR43) located just downstream from tRNA(Gly) genes. We confirmed that they are transcribed, probably from the tRNA gene promoter, producing dicistronic tRNA(Gly)-snoR43 precursors. Using transgenic lines expressing a tagged tRNA-snoR43.1 gene we show that the dicistronic precursor is accurately processed to both snoR43.1 and tRNA(Gly). In addition, we show that a recombinant RNase Z, the plant tRNA 3' processing enzyme, efficiently cleaves the dicistronic precursor in vitro releasing the snoR43.1 from the tRNA(Gly). Finally, we describe a similar case in rice implicating a tRNA(Met-e) expressed in fusion with a novel C/D snoRNA, showing that this mode of snoRNA expression is found in distant plant species. PMID- 12554663 TI - Low temperature inhibits RNA silencing-mediated defence by the control of siRNA generation. AB - Temperature dramatically affects plant-virus interactions. Outbreaks of virus diseases are frequently associated with low temperature, while at high temperature viral symptoms are often attenuated (heat masking) and plants rapidly recover from virus diseases. However, the underlying mechanisms of these well known observations are not yet understood. RNA silencing is a conserved defence system of eukaryotic cells, which operates against molecular parasites including viruses and transgenes. Here we show that at low temperature both virus and transgene triggered RNA silencing are inhibited. Therefore, in cold, plants become more susceptible to viruses, and RNA silencing-based phenotypes of transgenic plants are lost. Consistently, the levels of virus- and transgene derived small (21-26 nucleotide) interfering (si) RNAs-the central molecules of RNA silencing-mediated defence pathways-are dramatically reduced at low temperature. In contrast, RNA silencing was activated and the amount of siRNAs gradually increased with rising temperature. However, temperature does not influence the accumulation of micro (mi) RNAs, which play a role in developmental regulation, suggesting that the two classes of small (si and mi) RNAs are generated by different nuclease complexes. PMID- 12554664 TI - SMG-5, required for C.elegans nonsense-mediated mRNA decay, associates with SMG-2 and protein phosphatase 2A. AB - mRNAs that contain premature stop codons are degraded selectively and rapidly in eukaryotes, a phenomenon termed 'nonsense-mediated mRNA decay' (NMD). We report here molecular analysis of smg-5, which encodes a novel protein required for NMD in Caenorhabditis elegans. Using a combination of immunoprecipitation and yeast two-hybrid assays, we identified a series of protein-protein interactions involving SMG-5. SMG-5 interacts with at least four proteins: (i) SMG-7, a previously identified protein required for NMD; (ii) SMG-2, a phosphorylated protein required for NMD in worms, yeasts and mammals; (iii) PR65, the structural subunit of protein phosphatase 2A (PP2A); and (iv) PP2A(C), the catalytic subunit of PP2A. Previous work demonstrated that both SMG-5 and SMG-7 are required for efficient dephosphorylation of SMG-2. Our results suggest that PP2A is the SMG-2 phosphatase, and the role of SMG-5 is to direct PP2A to its SMG-2 substrate. We discuss cycles of SMG-2 phosphorylation and their roles in NMD. PMID- 12554665 TI - In vivo evidence for the prokaryotic model of extended codon-anticodon interaction in translation initiation. AB - Initiation codon context is an important determinant of translation initiation rates in both prokaryotes and eukaryotes. Such sequences include the Shine- Dalgarno ribosome-binding site, as well as other motifs surrounding the initiation codon. One proposed interaction is between the base immediately preceding the initiation codon (-1 position) and the nucleotide 3' to the tRNAf(Met) anticodon, at position 37. Adenine is conserved at position 37, and a uridine at -1 has been shown in vitro to favor initiation. We have tested this model in vivo, by manipulating the chloroplast of the green alga Chlamydomonas reinhardtii, where the translational machinery is prokaryotic in nature. We show that translational defects imparted by mutations at the petA -1 position can be suppressed by compensatory mutations at position 37 of an ectopically expressed tRNA(fMet). The mutant tRNAs are fully aminoacylated and do not interfere with the translation of other proteins. Although this extended base pairing is not an absolute requirement for initiation, it may convey added specificity to transcripts carrying non-standard initiation codons, and/or preserve translational fidelity under certain stress conditions. PMID- 12554666 TI - Wobble modification differences and subcellular localization of tRNAs in Leishmania tarentolae: implication for tRNA sorting mechanism. AB - In Leishmania tarentolae, all mitochondrial tRNAs are encoded in the nuclear genome and imported from the cytosol. It is known that tRNA(Glu)(UUC) and tRNA(Gln)(UUG) are localized in both cytosol and mitochondria. We investigated structural differences between affinity-isolated cytosolic (cy) and mitochondrial (mt) tRNAs for glutamate and glutamine by mass spectrometry. A unique modification difference in both tRNAs was identified at the anticodon wobble position: cy tRNAs have 5-methoxycarbonylmethyl-2- thiouridine (mcm(5)s(2)U), whereas mt tRNAs have 5- methoxycarbonylmethyl-2'-O-methyluridine (mcm(5)Um). In addition, a trace portion (4%) of cy tRNAs was found to have 5 methoxycarbonylmethyluridine (mcm(5)U) at its wobble position, which could represent a common modification intermediate for both modified uridines in cy and mt tRNAs. We also isolated a trace amount of mitochondria-specific tRNA(Lys)(UUU) from the cytosol and found mcm(5)U at its wobble position, while its mitochondrial counterpart has mcm(5)Um. Mt tRNA(Lys) and in vitro transcribed tRNA(Glu) were imported much more efficiently into isolated mitochondria than the native cy tRNA(Glu) in an in vitro importation experiment, indicating that cytosol-specific 2-thiolation could play an inhibitory role in tRNA import into mitochondria. PMID- 12554667 TI - Elucidation of tRNA-dependent editing by a class II tRNA synthetase and significance for cell viability. AB - Editing of misactivated amino acids by class I tRNA synthetases is encoded by a specialized internal domain specific to class I enzymes. In contrast, little is known about editing activities of the structurally distinct class II enzymes. Here we show that the class II alanyl-tRNA synthetase (AlaRS) has a specialized internal domain that appears weakly related to an appended domain of threonyl tRNA synthetase (ThrRS), but is unrelated to that found in class I enzymes. Editing of misactivated glycine or serine was shown to require a tRNA cofactor. Specific mutations in the aforementioned domain disrupt editing and lead to production of mischarged tRNA. This class-specific editing domain was found to be essential for cell growth, in the presence of elevated concentrations of glycine or serine. In contrast to ThrRS, where the editing domain is not found in all three kingdoms of living organisms, it was incorporated early into AlaRSs and is present throughout evolution. Thus, tRNA-dependent editing by AlaRS may have been critical for making the genetic code sufficiently accurate to generate the tree of life. PMID- 12554668 TI - ATP binding by glutamyl-tRNA synthetase is switched to the productive mode by tRNA binding. AB - Aminoacyl-tRNA synthetases catalyze the formation of an aminoacyl-AMP from an amino acid and ATP, prior to the aminoacyl transfer to tRNA. A subset of aminoacyl-tRNA synthetases, including glutamyl-tRNA synthetase (GluRS), have a regulation mechanism to avoid aminoacyl-AMP formation in the absence of tRNA. In this study, we determined the crystal structure of the 'non-productive' complex of Thermus thermophilus GluRS, ATP and L-glutamate, together with those of the GluRS.ATP, GluRS.tRNA.ATP and GluRS.tRNA.GoA (a glutamyl-AMP analog) complexes. In the absence of tRNA(Glu), ATP is accommodated in a 'non-productive' subsite within the ATP-binding site, so that the ATP alpha-phosphate and the glutamate alpha-carboxyl groups in GluRS. ATP.Glu are too far from each other (6.2 A) to react. In contrast, the ATP-binding mode in GluRS.tRNA. ATP is dramatically different from those in GluRS.ATP.Glu and GluRS.ATP, but corresponds to the AMP moiety binding mode in GluRS.tRNA.GoA (the 'productive' subsite). Therefore, tRNA binding to GluRS switches the ATP-binding mode. The interactions of the three tRNA(Glu) regions with GluRS cause conformational changes around the ATP-binding site, and allow ATP to bind to the 'productive' subsite. PMID- 12554669 TI - The proline-rich homeodomain protein, PRH, is a tissue-specific inhibitor of eIF4E-dependent cyclin D1 mRNA transport and growth. AB - The translation initiation factor eIF4E is involved in the modulation of cellular growth. In the nucleus, where eIF4E is associated with PML nuclear bodies, eIF4E mediates nucleocytoplasmic transport of specific transcripts, and this contributes to its transformation activity. Surprisingly, we found that a trans cription factor, the proline-rich homeodomain protein PRH, is a negative regulator of eIF4E in myeloid cells, interacting with eIF4E through a conserved binding site typically found in translational regulators. Through this interaction, PRH inhibits eIF4E-dependent mRNA transport and subsequent transformation. These activities of PRH are independent of its transcriptional functions. Further, we found that 199 homeodomain proteins contain potential eIF4E-binding sites. Thus, there could be many tissue-specific regulators of eIF4E. These findings provide a model for regulation of a general factor, eIF4E, in tissue- specific contexts, and suggest that its regulation is important in differentiation and development. PMID- 12554670 TI - Human replication protein Cdc6 prevents mitosis through a checkpoint mechanism that implicates Chk1. AB - In yeasts, the replication protein Cdc6/Cdc18 is required for the initiation of DNA replication and also for coupling S phase with the following mitosis. In metazoans a role for Cdc6 has only been shown in S phase entry. Here we provide evidence that human Cdc6 (HuCdc6) also regulates the onset of mitosis, as overexpression of HuCdc6 in G(2) phase cells prevents entry into mitosis. This block is abolished when HuCdc6 is expressed together with a constitutively active Cyclin B/CDK1 complex or with Cdc25B or Cdc25C. An inhibitor of Chk1 kinase activity, UCN-01, overcomes the HuCdc6 mediated G(2) arrest indicating that HuCdc6 blocks cells in G(2) phase via a checkpoint pathway involving Chk1. When HuCdc6 is overexpressed in G(2), we detected phosphorylation of Chk1. Thus, HuCdc6 can trigger a checkpoint response, which could ensure that all DNA is replicated before mitotic entry. We also present evidence that the ability of HuCdc6 to block mitosis may be regulated by its phosphorylation. PMID- 12554671 TI - Chk1-deficient tumour cells are viable but exhibit multiple checkpoint and survival defects. AB - The conserved protein kinase Chk1 is believed to play an important role in checkpoint responses to aberrant DNA structures; however, genetic analysis of Chk1 functions in metazoans is complicated by lethality of Chk1-deficient embryonic cells. We have used gene targeting to eliminate Chk1 function in somatic DT40 B-lymphoma cells. We find that Chk1-deficient DT40 cells are viable, but fail to arrest in G(2)/M in response to and are hypersensitive to killing by ionizing radiation. Chk1-deficient cells also fail to maintain viable replication forks or suppress futile origin firing when DNA polymerase is inhibited, leading to incomplete genome duplication and diminished cell survival after release from replication arrest. In contrast to embryonic cells, however, Chk1 is not required to delay mitosis when DNA synthesis is inhibited. Thus, Chk1 is dispensable for normal cell division in somatic DT40 cells but is essential for DNA damage induced G(2)/M arrest and a subset of replication checkpoint responses. Furthermore, Chk1-dependent processes promote tumour cell survival after perturbations of DNA structure or metabolism. PMID- 12554672 TI - A model for dsDNA translocation revealed by a structural motif common to RecG and Mfd proteins. AB - RecG protein differs from other helicases analysed to atomic resolution in that it mediates strand separation via translocation on double-stranded (ds) rather than single-stranded (ss) DNA. We describe a highly conserved helical hairpin motif in RecG and show it to be important for helicase activity. It places two arginines (R609 and R630) in opposing positions within the component helices where they are stabilized by a network of hydrogen bonds involving a glutamate from helicase motif VI. We suggest that disruption of this feature, triggered by ATP hydrolysis, moves an adjacent loop structure in the dsDNA-binding channel and that a swinging arm motion of this loop drives translocation. Substitutions that reverse the charge at R609 or R630 reduce DNA unwinding and ATPase activities, and increase dsDNA binding, but do not affect branched DNA binding. Sequences forming the helical hairpin and loop structures are highly conserved in Mfd protein, a transcription-coupled DNA repair factor that also translocates on dsDNA. The possibility of type I restriction enzymes and chromatin-remodelling factors using similar structures to drive translocation on dsDNA is discussed. PMID- 12554673 TI - The RdgC protein of Escherichia coli binds DNA and counters a toxic effect of RecFOR in strains lacking the replication restart protein PriA. AB - PriA protein provides a means to load the DnaB replicative helicase at DNA replication fork and D loop structures, and is therefore a key factor in the rescue of stalled or broken forks and subsequent replication restart. We show that the nucleoid-associated RdgC protein binds non-specifically to single stranded (ss) DNA and double-stranded DNA. It is also essential for growth of a strain lacking PriA, indicating that it might affect replication fork progression or fork rescue. dnaC suppressors of priA overcome this inviability, especially when RecF, RecO or RecR is inactivated, indicating that RdgC avoids or counters a toxic effect of these proteins. Mutations modifying ssDNA-binding (SSB) protein also negate this toxic effect, suggesting that the toxicity reflects inappropriate loading of RecA on SSB-coated ssDNA, leading to excessive or untimely RecA activity. We suggest that binding of RdgC to DNA limits RecA loading, avoiding problems at replication forks that would otherwise require PriA to promote replication restart. Mutations in RNA polymerase also reduce the toxic effect of RecFOR, providing a further link between DNA replication, transcription and repair. PMID- 12554675 TI - Synonymous mutations in the human dopamine receptor D2 (DRD2) affect mRNA stability and synthesis of the receptor. AB - Although changes in nucleotide sequence affecting the composition and the structure of proteins are well known, functional changes resulting from nucleotide substitutions cannot always be inferred from simple analysis of DNA sequence. Because a strong synonymous codon usage bias in the human DRD2 gene, suggesting selection on synonymous positions, was revealed by the relative independence of the G+C content of the third codon positions from the isochoric G+C frequencies, we chose to investigate functional effects of the six known naturally occurring synonymous changes (C132T, G423A, T765C, C939T, C957T, and G1101A) in the human DRD2. We report here that some synonymous mutations in the human DRD2 have functional effects and suggest a novel genetic mechanism. 957T, rather than being 'silent', altered the predicted mRNA folding, led to a decrease in mRNA stability and translation, and dramatically changed dopamine-induced up regulation of DRD2 expression. 1101A did not show an effect by itself but annulled the above effects of 957T in the compound clone 957T/1101A, demonstrating that combinations of synonymous mutations can have functional consequences drastically different from those of each isolated mutation. C957T was found to be in linkage disequilibrium in a European-American population with the -141C Ins/Del and TaqI 'A' variants, which have been reported to be associated with schizophrenia and alcoholism, respectively. These results call into question some assumptions made about synonymous variation in molecular population genetics and gene-mapping studies of diseases with complex inheritance, and indicate that synonymous variation can have effects of potential pathophysiological and pharmacogenetic importance. PMID- 12554674 TI - The alternating ATPase domains of MutS control DNA mismatch repair. AB - DNA mismatch repair is an essential safeguard of genomic integrity by removing base mispairings that may arise from DNA polymerase errors or from homologous recombination between DNA strands. In Escherichia coli, the MutS enzyme recognizes mismatches and initiates repair. MutS has an intrinsic ATPase activity crucial for its function, but which is poorly understood. We show here that within the MutS homodimer, the two chemically identical ATPase sites have different affinities for ADP, and the two sites alternate in ATP hydrolysis. A single residue, Arg697, located at the interface of the two ATPase domains, controls the asymmetry. When mutated, the asymmetry is lost and mismatch repair in vivo is impaired. We propose that asymmetry of the ATPase domains is an essential feature of mismatch repair that controls the timing of the different steps in the repair cascade. PMID- 12554676 TI - Tissue-specific RNA surveillance? Nonsense-mediated mRNA decay causes collagen X haploinsufficiency in Schmid metaphyseal chondrodysplasia cartilage. AB - Mutations resulting in a premature termination codon (PTC) are a major cause of inherited disorders, and the majority of these mutant RNA transcripts are subjected to nonsense-mediated mRNA decay (NMD). This RNA surveillance results in reduced mutant allele expression, the extent of which can impact on the clinical severity. The molecular mechanisms of NMD in mammalian cells, its relationship to splicing and translation, downstream sequence elements and binding factors remains only partially understood. Currently there is little information on whether the extent of NMD is gene- or tissue-specific, although nonsense mutation inhibition of RNA splicing has been shown to exhibit some tissue and gene specificity in vitro. Schmid metaphyseal chondrodysplasia results from heterozygous mutations in the gene for collagen X (COL10A1), expressed by the hypertrophic chondrocytes of growth plate cartilage. In one patient a PTC mutation has been shown to result in complete NMD and collagen X haploinsufficiency in cartilage. Here we show that, in this patient, and in another with a different collagen X PTC mutation also leading to complete NMD in cartilage, the mutant mRNAs were not subjected to NMD in non-cartilage cells (lymphoblasts and bone cells). These data suggest that novel RNA surveillance mechanisms may exist in cartilage and that tissue specificity of NMD could be of importance in understanding the molecular pathology of nonsense mutations. Furthermore, the demonstration of collagen X haploinsufficiency in the second patient to be studied at the level of tissue expression, confirms that nonsense mutations leading to complete mutant collagen X mRNA degradation in cartilage is an important molecular cause of SMCD. PMID- 12554677 TI - Role of oxidative stress in telomere shortening in cultured fibroblasts from normal individuals and patients with ataxia-telangiectasia. AB - Cells from patients with the autosomal recessive disorder ataxia-telangiectasia (A-T) display accelerated telomere shortening upon culture in vitro. It has been suggested that A-T cells are in a chronic state of oxidative stress, which could contribute to their enhanced telomere shortening. In order to examine this hypothesis, we monitored the changes in telomere length in A-T homozygous, heterozygous and control fibroblasts cultured in vitro under various conditions of oxidative stress using quantitative fluorescent in situ hybridization. Compared with normal cells, the rate of telomere shortening was 1.5-fold increased under 'normal' levels of oxidative stress in A-T heterozygous cells and 2.4-3.2-fold in A-T homozygous cells. Mild chronic oxidative stress induced by hydrogen peroxide increased the rate of telomere shortening in A-T cells but not in normal fibroblasts and the telomere shortening rate decreased in both normal and A-T fibroblasts if cultures were supplemented with the anti-oxidant phenyl butyl-nitrone. Increased telomere shortening upon oxidative stress in A-T cells was associated with a significant increase in the number of extra-chromosomal fragments of telomeric DNA and chromosome ends without detectable telomere repeats. We propose that the ATM (A-T mutated) protein has a role in the prevention or repair of oxidative damage to telomeric DNA and that enhanced sensitivity of telomeric DNA to oxidative damage in A-T cells results in accelerated telomere shortening and chromosomal instability. PMID- 12554679 TI - ETS2 overexpression in transgenic models and in Down syndrome predisposes to apoptosis via the p53 pathway. AB - ETS2 is a transcription factor encoded by a gene on human chromosome 21 and alterations in its expression have been implicated in the pathophysiological features of Down syndrome (DS). This study demonstrates that overexpression of ETS2 results in apoptosis. This is shown in a number of circumstances, including ETS2-overexpressing transgenic mice and cell lines and in cells from subjects with DS. Indeed we report for the first time that the ETS2 overexpression transgenic mouse develops a smaller thymus and lymphocyte abnormalities similar to that observed in DS. In all circumstances of ETS2 overexpression, the increased apoptosis correlated with increased p53 and alterations in downstream factors in the p53 pathway. In the human HeLa cancer cell line, transfection with functional p53 enables ETS2 overexpression to induce apoptosis. Furthermore, crossing the ETS2 transgenic mice with p53(-/-) mice genetically rescued the thymic apoptosis phenotype. Therefore, we conclude that overexpression of human chromosome 21-encoded ETS2 induces apoptosis that is dependent on p53. These results have important consequences for understanding DS and oncogenesis and may provide new insights into therapeutic interventions. PMID- 12554680 TI - Defective endocytic trafficking of NPC1 and NPC2 underlying infantile Niemann Pick type C disease. AB - Niemann-Pick type C (NPC) disease is a fatal recessively inherited lysosomal cholesterol-sphingolipidosis. Mutations in the NPC1 gene cause approximately 95% of the cases, the rest being caused by NPC2 mutations. Here the molecular basis of a severe infantile form of the disease was dissected. The level of NPC1 protein in the patient fibroblasts was similar to that in control cells. However, the protein was partially mislocalized from late endocytic organelles diffusely to the cell periphery. In contrast, NPC2 was upregulated and accumulated in cholesterol storing late endocytic organelles. Two point mutations and a four nucleotide deletion were identified in the NPC1 gene, leading to the amino acid substitutions C113R, P237S and deletion of 37 C-terminal amino acids (delC). Overexpression of individual NPC1 mutations revealed that delC produced an unstable protein, wild-type and NPC1-P237S colocalized with Rab7-positive late endosomes whereas NPC1-C113R localized to the ER, Rab7-negative endosomes and the cell surface. Expression of wild-type or NPC1-P237S cleared the lysosomal cholesterol accumulation in NPC1-deficient cells whereas C113R or delC did not. In the Finnish and Swedish population samples, alleles carrying C113R or delC were not identified, whereas approximately 5% of the alleles carried P237S. Our studies identify P237S as a prevalent NPC1 polymorphism and delC and C113R as deleterious NPC1 mutations. Moreover, they show that delC leads to rapid degradation of NPC1 and C113R to endocytic missorting of the protein. These changes are accompanied by lysosomal accumulation of NPC2, suggesting that NPC1 governs the endocytic transport of NPC2. PMID- 12554681 TI - Mismatch repair gene Msh2 modifies the timing of early disease in Hdh(Q111) striatum. AB - Somatic instability of expanded HD CAG repeats that encode the polyglutamine tract in mutant huntingtin has been implicated in the striatal selectivity of Huntington's disease (HD) pathology. Here in Hdh(Q111) mice, we have tested whether a genetic background deficient in Msh2, expected to eliminate the unstable behavior of the 109 CAG array inserted into the murine HD gene, would alter the timing or striatal specificity of a dominant disease phenotype that predicts late-onset neurodegeneration. Our analyses of Hdh(Q111/+):Msh2(+/+) and Hdh(Q111/+): Msh2(-/-) progeny revealed that, while inherited instability involved Msh2-dependent and -independent mechanisms, lack of Msh2 was sufficient to abrogate progressive HD CAG repeat expansion in striatum. The absence of Msh2 also eliminated striatal mutant huntingtin with somatically expanded glutamine tracts and caused an approximately 5 month delay in nuclear mutant protein accumulation, but did not alter the striatal specificity of this early phenotype. Thus, somatic HD CAG instability appears to be a consequence of a striatal selective disease process that accelerates the timing of an early disease phenotype, via expansion of the glutamine tract in mutant huntingtin. Therefore Msh2, as a striking modifier of early disease onset in a precise genetic HD mouse model, provides a novel target for the development of pharmacological agents that aim to slow pathogenesis in man. PMID- 12554682 TI - A differentially methylated region within the gene Kcnq1 functions as an imprinted promoter and silencer. AB - The imprinted gene cluster on mouse distal chromosome 7 contains a differentially methylated CpG island that maps within the Kcnq1 gene that has been shown to be required for the imprinting of multiple genes. To evaluate models for how this imprinting control region (ICR) regulates imprinting, we have characterized it structurally and functionally. We show that the region contains a promoter for a paternally expressed anti-sense transcript, Kcnq1ot1, and we define the extent of the minimal promoter. We describe three paternal-specific nuclease hypersensitive sites immediately upstream from the start site and show that they are required for full promoter activity. The expression of Kcnq1ot1 during pre- and postnatal development is compared to that of other imprinted genes in its vicinity, Cdnkn1c and Kcnq1. The lack of coordination in their expression tends to rule out an enhancer competition model for the action of the ICR in imprinting control. Using a stable transfection assay we show that the region contains a position independent and orientation-independent silencer. We propose, on the basis of these findings, that the Kcnq1 ICR functions as a silencer on the paternal chromosome to effect the repression of neighboring genes. PMID- 12554683 TI - Epigenetic modifications in an imprinting cluster are controlled by a hierarchy of DMRs suggesting long-range chromatin interactions. AB - Imprinted genes and their control elements occur in clusters in the mammalian genome and carry epigenetic modifications. Observations from imprinting disorders suggest that epigenetic modifications throughout the clusters could be under regional control. However, neither the elements that are responsible for regional control, nor its developmental timing, particularly whether it occurs in the germline or postzygotically, are known. Here we examine regional control of DNA methylation in the imprinted Igf2-H19 region in the mouse. Paternal germline specific methylation was reprogrammed after fertilization in two differentially methylated regions (DMRs) in Igf2, and was reestablished after implantation. Using a number of knockout strains in the region, we found that the DMRs themselves are involved in regional coordination in a hierarchical fashion. Thus the H19 DMR was needed on the maternal allele to protect the Igf2 DMRs 1 and 2 from methylation, and Igf2 DMR1 was needed to protect DMR2 from methylation. This regional coordination occurred exclusively after fertilization during somatic development, and did not involve linear spreading of DNA methylation, suggesting a model in which long-range chromatin interactions are involved in regional epigenetic coordination. These observations are likely to be relevant to other gene clusters in which epigenetic regulation plays a role, and in pathological situations in which epigenetic regulation is disrupted. PMID- 12554684 TI - Suppression of polyglutamine-induced protein aggregation in Caenorhabditis elegans by torsin proteins. AB - Torsion dystonia is an autosomal dominant movement disorder characterized by involuntary, repetitive muscle contractions and twisted postures. The most severe early-onset form of dystonia has been linked to mutations in the human DYT1 (TOR1A) gene encoding a protein termed torsinA. While causative genetic alterations have been identified, the function of torsin proteins and the molecular mechanism underlying dystonia remain unknown. Phylogenetic analysis of the torsin protein family indicates these proteins share distant sequence similarity with the large and diverse family of AAA+ proteins. We have established the nematode, Caenorhabditis elegans, as a model system for examining torsin activity. Using an in vivo assay for polyglutamine repeat-induced protein aggregation in living animals, we have determined that ectopic overexpression of both human and C. elegans torsin proteins results in a dramatic reduction of polyglutamine-dependent protein aggregation in a manner similar to that previously reported for molecular chaperones. The suppressive effects of torsin overexpression persisted as animals aged, whereas a mutant nematode torsin protein was incapable of ameliorating aggregate formation. Antibody staining of transgenic animals indicated that both the C. elegans torsin-related protein TOR 2 and ubiquitin were localized to sites of protein aggregation. These data represent the first functional evidence of a role for torsins in effectively managing protein folding and suggest that possible breakdown in a neuroprotective mechanism that is, in part, mediated by torsins may be responsible for the neuronal dysfunction associated with dystonia. PMID- 12554685 TI - An isoform of hPANK2, deficient in pantothenate kinase-associated neurodegeneration, localizes to mitochondria. AB - Mutations in the human PANK2 gene have been shown to occur in autosomal-recessive pantothenate kinase-associated neurodegeneration, a syndrome originally described by Hallervorden and Spatz. The kinase catalyses the first and rate-limiting step in the biosynthesis of coenzyme A, a key molecule in energy metabolism. We have determined the exon-intron structure of the hPANK2 gene and identified two alternatively used first exons. The resulting transcripts encode distinct isoforms of hPANK2, one of which carries an N-terminal extension with a predicted mitochondrial targeting signal. An in vitro import assay and in vivo immunolocalization experiments demonstrate a mitochondrial localization of this isoform. We conclude that the symptoms observed in pantothenate kinase-associated neurodegeneration are caused by a deficiency of the mitochondrial isoform and postulate the existence of a complete intramitochondrial pathway for de novo synthesis of coenzyme A. PMID- 12554678 TI - Methylation-sensitive binding of transcription factor YY1 to an insulator sequence within the paternally expressed imprinted gene, Peg3. AB - The 5'-ends of two paternally expressed mouse genes, Peg3 and Usp29, are jointly associated with a CpG island that exhibits allele-specific methylation. Sequence comparison of the regions derived from human, mouse and cow revealed the presence of two evolutionarily conserved sequence motifs including one that is repeated multiple times within the first intron of Peg3 in all three mammals. DNA mobility shift and chromatin immunoprecipitation (ChIP) assays clearly demonstrated that this motif is an in vivo binding site for the Gli-type transcription factor YY1. The YY1-binding site contains one CpG dinucleotide, and methylation of this CpG site abolishes the binding activity of YY1 in vitro. The Peg3 YY1-binding sites are methylated only on the maternal chromosome in vivo, and ChIP assays confirmed that YY1 binds specifically to the paternal allele of the gene. Promoter, enhancer and insulator assays with deletion constructs of sequence surrounding the YY1-binding sites indicate that the region functions as a methylation sensitive insulator that may influence the imprinted expression of Peg3 and neighboring genes. The current study is the first report demonstrating the involvement of YY1 in methylation-sensitive insulator activity and suggests a potential role of this highly conserved protein in mammalian genomic imprinting. PMID- 12554686 TI - An intragenic suppressor in the cytochrome c oxidase I gene of mouse mitochondrial DNA. AB - We report here the identification of a cell line containing single and double missense mutations in cytochrome c oxidase (COX) subunit I gene of mouse mitochondrial DNA. When present in homoplasmy, the single mutant displays a normal complex IV assembly but a significantly reduced COX activity, while the double mutant almost completely compensates the functional defect of the first mutation. We discuss the potential structural consequences of those mutations based on the modeled structure of mouse complex IV. Based on genetic, biochemical and molecular analyses of cultured mouse cells we infer that: (i) deleterious mutations can arise and become predominant; (ii) cultured cells can maintain several mtDNA haplotypes at stable frequencies; (iii) the respiratory chain has little spare COX capacity; and (iv) the size of a cavity in the vicinity of Val421 in CO I of animal COX may affect the function of the enzyme. PMID- 12554688 TI - Mutation of the SBF2 gene, encoding a novel member of the myotubularin family, in Charcot-Marie-Tooth neuropathy type 4B2/11p15. AB - Autosomal recessive hereditary motor and sensory neuropathy or Charcot-Marie Tooth disease (CMT) is a severe childhood-onset neuromuscular disorder. Autosomal recessive CMT is genetically heterogeneous with one locus mapped to chromosome 11p15 (CMT4B2). The histopathological hallmarks of CMT4B2 are focal outfoldings of myelin in nerve biopsies. Homozygosity mapping, in a Turkish inbred family with four children affected by CMT characterized by focally folded myelin, provided linkage to the CMT4B2 locus. We identified a large, novel gene, named SET binding factor 2 (SBF2), that lies within this interval and is expressed in various tissues, including spinal cord and peripheral nerve. SBF2 is a member of the pseudo-phosphatase branch of myotubularins and was an obvious candidate for CMT4B2 by virtue of its striking homology to myotubularin-related protein 2 (MTMR2), causing another form of autosomal recessive CMT with outfoldings of the myelin sheaths. Molecular study of the SBF2 gene in the CMT4B family demonstrated the presence of a homozygous inframe deletion of SBF2 exons 11 and 12 in all four affected individuals. On the protein level, this mutation is predicted to disrupt an N-terminal domain that is conserved in SBF2 and its orthologues across species. Myotubularin-related proteins have been suggested to work in phosphoinositide-mediated signalling events that may also convey control of myelination. Localization of SBF2 within the candidate interval, cosegregation with the disease, expression in the peripheral nervous system, and resemblance of the histopathological phenotype to that related to mutations in its paralogue MTMR2 indicate that this gene is the CMT4B2 gene. PMID- 12554687 TI - Duplications of the AZFa region of the human Y chromosome are mediated by homologous recombination between HERVs and are compatible with male fertility. AB - Deletions of the AZFa region on the long arm of the human Y chromosome cause male infertility. Previous work has shown that this is an example of a genomic disorder, since most deletions are caused by non-allelic homologous recombination between endogenous retroviral elements (HERVs) flanking the 780 kb region. The reciprocal products of these deletion events, AZFa duplications, have not been reported to date. Here we show that duplication chromosomes exist in population samples by detecting Y-chromosomal short tandem repeat (YSTR) allele duplications within the AZFa region, and by showing that two chromosomes carrying these duplicated alleles contain a third junction-specific HERV sequence. Sequence analysis of these cases, which most likely represent independent duplication events, shows that breakpoints lie in the same region of inter-HERV sequence identity as do deletion breakpoints, and thus that the mechanism of duplication is indeed the reciprocal of deletion. Consideration of the accumulated Y-STR allele diversity between duplicated copies of the AZFa region indicates that one of the duplication chromosomes has been in the population for at least 17 generations, and therefore must be compatible with male fertility. PMID- 12554689 TI - Mutations in a novel gene Dymeclin (FLJ20071) are responsible for Dyggve-Melchior Clausen syndrome. AB - Dyggve-Melchior-Clausen syndrome (DMC) is a rare autosomal-recessive disorder, the gene for which maps to chromosome 18q21.1. DMC is characterized by the association of a spondylo-epi-metaphyseal dysplasia and mental retardation. Electron microscopic study of cutaneous cells of an affected child showed dilated rough endoplasmic reticulum, enlarged and aberrant vacuoles and numerous vesicles. As the etiology of the disorder is unknown, we have used a positional cloning strategy to identify the DMC gene. We detected seven deleterious mutations within a gene predicted from a human transcript (FLJ20071) in 10 DMC families. The mutations were nonsense mutations (R194X, R204X, L219X, Q483X), splice site or frameshift mutations (K626N+92aa to stop). The DMC gene transcript is widely distributed but appears abundant in chondrocytes and fetal brain. The predicted protein product of the DMC gene yields little insight into its likely function, showing no significant homology to any known protein family. However, the carboxy terminal end comprises a cluster of dileucine motifs, highly conserved across species. We conclude that DMC syndrome is consequent upon loss of function of a gene that we propose to name Dymeclin, which may have a role in process of intracellular digestion of proteins. PMID- 12554690 TI - When east meets west: the relationship between yin-yang and antioxidation oxidation. AB - Ancient traditional Chinese medicine (TCM) has effectively relied on the theory of yin-yang balance in diagnoses and treatments of diseases and disorders for more than 2000 years. However, in eastern society, yin-yang is regarded as an incomprehensible ideology without definite physical meaning. Consequently, the yin-yang balance in medicine has not been studied by modern scientific means. In the western world, yin-yang balance is often misunderstood as a religious belief or a principle of lifestyle. Herein, we attempted to define the physical meaning of yin-yang in TCM by correlating it with biochemical processes. We propose that yin-yang balance is antioxidation-oxidation balance with yin representing antioxidation and yang as oxidation. Our proposal is partially supported by the fact that the yin-tonic traditional Chinese herbs have, on average, about six times more antioxidant activity and polyphenolic contents than the yang-tonic herbs. Our hypothesis opens an avenue to systematically study the yin-yang balance and its health implications with the use of modern biochemical tools. PMID- 12554691 TI - SNAREs and control of synaptic release probabilities. AB - Since quantal release was first described, it has been clear that release of neurotransmitters is a stochastic process. Modulation of neurotransmitter release probability by regulatory factors likely affects the transfer of information within the nervous system. Although many rules governing release probabilities at the synapse have been discovered, their molecular basis is still under investigation. Here we analyze stimulus-evoked probabilistic assembly of the SNARE fusion machinery and show that a simple SNARE-based mechanism can account quantitatively for the classical binomial behavior of stochastic neurotransmitter release. Our analysis highlights for the first time how the fusion machinery, which is directly responsible for neurotransmitter release, may also contribute to the rich variety of synaptic responses. PMID- 12554692 TI - Autoantibodies to the epidermal growth factor receptor in systemic sclerosis, lupus, and autoimmune mice. AB - Autoantibodies to the recombinant extracellular domain of epidermal growth factor receptor (exEGFR) were detected by ELISA in the serum of Fas-defective old MRL/MpJ/lpr and C3H/HeJ/gld mice, but not young mice from these strains, or nonautoimmune young and old BALB/c, MRL/MpJ/++, and C3H/HeJ/MMTV mice. Compared with control human subjects without autoimmune disease, the frequency of exEGFR binding autoantibodies was increased in scleroderma (systemic sclerosis) patients and to a lesser extent in lupus patients. Phage autoantibodies (Fv fragments) isolated from a lupus library by selection on a linear epitope of EGFR (residues 294-310) displayed the ability to bind exEGFR. Treatment of EGFR-expressing A431 cells with autoantibodies purified by affinity chromatography on immobilized exEGFR resulted in specific staining of the cells. Short-lived but strong inhibition of cellular DNA synthesis was observed in the presence of the autoantibodies. We concluded that autoantibody responses to EGFR hold the potential of fulfilling a pathogenic role in autoimmune disease. PMID- 12554693 TI - Novel regulation of cardiac force-frequency relation by CREM (cAMP response element modulator). AB - The cAMP response element modulator (CREM) plays pivotal roles in the hypothalamic-pituitary-gonadal axis. CREM mRNA is robustly expressed in human myocardium, and identified isoforms may suppress cAMP response element-mediated transcription. However, little is known about the physiological importance of CREM in intact hearts remains unknown. We studied CREM-null mice and age-matched control littermates by in vivo pressure-volume loops to analyze basal and reserve cardiac function. Basal systolic and diastolic function, echocardiographic morphology, and myocardial histology were normal in CREM-null animals. However functional reserve with increasing heart rate was markedly depressed, with less contractile augmentation (+22+/-9% CREM-/- vs.+62+/-11% controls, P<0.05) and relaxation shortening (5+/-5% CREM-/- vs. -18+/-3% controls; P<0.05) at faster rates. In contrast, isoproterenol dose-responses were similar, suggesting normal beta-adrenergic receptor-coupled signaling. Gene expression of calcium handling proteins (SERCA, phospholamban) and stress-response genes (e.g., alpha-skeletal actin, beta-myosin heavy chain, natriuretic peptides) were similar between groups. However, total and serine-phosphorylated phospholamban protein declined 38 and -64% respectively, and protein phosphatase-1 (PP1) activity increased 44% without increased protein levels (all P<0.01) in CREM-/- vs. controls. These results demonstrate novel involvement of CREM in regulation of PP1 activity and of PLB, likely resulting in a potent frequency-dependent influence on cardiac function. PMID- 12554694 TI - Induction of a p95/Nbs1-mediated S phase checkpoint by telomere 3' overhang specific DNA. AB - Telomere shortening induces a nonproliferative senescent phenotype, believed to reduce cancer risk, and telomeres are involved in a poorly understood manner in responses to DNA damage. Although telomere disruption induces p53 and triggers apoptosis or cell cycle arrest, the features of the disrupted telomere that trigger this response and the precise mechanism involved are poorly understood. Using human cells, we show that DNA oligonucleotides homologous to the telomere 3' overhang sequence specifically induce and activate p53 and activate an S phase checkpoint by modifying the Nijmegen breakage syndrome protein, known to mediate the S phase checkpoint after DNA damage. These responses are mediated, at least in part, by the ATM kinase and are not attributable to disruption of cellular telomeres. Based on these and earlier data, we propose that these oligonucleotides mimic a physiological signal, exposure of the telomere 3' overhang due to opening of the normal telomere loop structure, and hence evoke these protective antiproliferative responses in the absence of DNA damage or telomere disruption. PMID- 12554695 TI - Oncogenic transformation induces tumor angiogenesis: a role for PAR1 activation. AB - The formation of new blood vessels is a critical determinant of tumor progression. We find that Par1 gene expression plays a central role in blood vessel recruitment in animal models. By in vivo injection of either Matrigel plugs containing Par1-expressing cells or of rat prostatic carcinoma cells transfected with tetracycline-inducible Par1 expression vectors, we show that Par1 significantly enhances both angiogenesis and tumor growth. Several vascular endothelial growth factor (VEGF) splice forms are induced in cells expressing Par1. Activation of PAR1 markedly augments the expression of VEGF mRNAs and of functional VEGFs as determined by in vitro assays for endothelial tube alignment and bovine aortic endothelial cell proliferation. Because neutralizing anti-VEGF antibodies potently inhibited Par1-induced endothelial cell proliferation, we conclude that Par1-induced angiogenesis requires VEGF. Specific inhibitors of protein kinase C (PKC), Src, and phosphatidylinositol 3-kinase (PI3K) inhibit Par1-induced VEGF expression, suggesting the participation of these kinases in the process. We also show that oncogenic transformation by genes known to be part of PAR1 signaling machinery is sufficient to increase VEGF expression in NIH 3T3 cells. These data support the novel notion that initiation of cell signaling either by activating PAR1 or by the activated forms of oncogenes is sufficient to induce VEGF and hence angiogenesis. PMID- 12554696 TI - Heme oxygenase and nitric oxide synthase mediate cooling-associated protection against TNF-alpha-induced microcirculatory dysfunction and apoptotic cell death. AB - Local cooling protects against TNF-alpha-induced injury by attenuating inflammation-associated microcirculatory dysfunction and leukocytic response. Mechanisms of protection, however, are not fully understood. We studied whether the metabolites of the HO and NOS pathway, exerting potent vasodilatory, antioxidant, and anti-apoptotic properties, are involved in tissue cryoprotection. In animals pretreated with L-NAME or SnPP-IX, cooling-associated abrogation of TNF-alpha-induced microcirculatory dysfunction was abolished. Combined L-NAME/SnPP-IX pretreatment did not cause greater blunting than seen when each mediator system was inhibited separately. In SnPP-IX- but not L-NAME pretreated animals, transient hypothermia failed to reduce TNF-alpha-mediated leukocyte adherence. Vice versa, treatment of TNF-alpha-exposed animals with either the NO donor l-arginine or the HO-1 inductor hemin mimicked cooling associated tissue protection except for failure of l-arginine to abrogate the inflammatory leukocyte response. The efficiency of cooling to inhibit TNF-alpha induced apoptotic cell death was blunted in SnPP-IX-, L-NAME-, and SnPP-IX/L-NAME pretreated animals. Coadministration of Trolox in SnPP-IX-treated animals partly attenuated leukocyte adherence and cell apoptosis, implying that the HO pathway metabolite biliverdin contributes to the salutary effects of cooling. Thus, our study provides evidence that metabolites of the HO and the NOS pathway mediate the cooling-associated protection of inflamed tissue. Biliverdin rather than CO and NO mediates the anti-inflammatory action, whereas a coordinated function of the gaseous monoxides prevents microcirculatory dysfunction and apoptotic cell death. PMID- 12554697 TI - Vascular endothelial growth factor promotes proliferation of cortical neuron precursors by regulating E2F expression. AB - Neurogenesis, or the production of new neurons, is regulated by physiological and pathological processes including aging, stress, and brain injury. Many mitogenic and trophic factors that regulate proliferation of nonneuronal cells are also involved in neurogenesis. These include vascular endothelial cell growth factor (VEGF), which stimulates the incorporation of bromodeoxyuridine (BrdU) into neuronal precursor cells in vitro and in the adult rat brain in vivo. Using BrdU labeling as an index of cell proliferation, we found that the in vitro neuroproliferative effect of VEGF was associated with up-regulation of E2F family transcription factors, cyclin D1, cyclin E, and cdc25. VEGF also increased nuclear expression of E2F1, E2F2, and E2F3, consistent with regulation of the G1/S phase transition of the cell cycle. The proliferative effect of VEGF was inhibited by the extracellular signal-regulated kinase kinase (MEK) inhibitor PD98059, the phospholipase C (PLC) inhibitor U73122, the protein kinase C (PKC) inhibitor GF102390X, and the phosphatidylinositol 3-kinase (PI3K) inhibitor wortmannin, indicating involvement of multiple signaling pathways. These findings help to provide a molecular basis for some of the recently identified neuronal effects of VEGF. PMID- 12554698 TI - SARA and Hgs attenuate susceptibility to TGF-beta1-mediated T cell suppression. AB - Transforming growth factor-beta1 (TGF-beta1) is a pluripotent cytokine that controls peripheral T cell tolerance mainly in mucosal immunity. It is secreted by regulatory T cells (Tr /Th3) but also by other immununologically active cells. Smad anchor for receptor activation (SARA) and hepatic growth factor-regulated tyrosine kinase substrate (Hgs) are involved in TGF-beta1 signaling. Both molecules are known to present Smad2 and Smad3 to the TGF-beta receptor complex. The role of SARA and Hgs in TGF-beta1 susceptibility of human CD4+ T cells is unclear. We demonstrate here that TGF-beta1 up-regulates SARA mRNA expression in CD4+ T cells similar to that of Smad7. However, the increase in SARA expression was lower (6.1+/-0.3-fold vs. 25+/-4.1-fold) compared with Smad7 and delayed, with a maximum at 12 h compared with 2 h. Th1 and Th2 cell subsets expressed the same levels of SARA and Hgs. Compared with resting cells, significantly lower levels of the two molecules were found in antigen/allergen- or anti-CD3/CD28 stimulated cells. Down-regulation of SARA and Hgs mRNA in preactivated CD4+ T cells was accompanied by a twofold increase in a TGF-beta1 responsive reporter gene assay. Overexpression of SARA and Hgs in T cells yielded a dose-dependent decrease in cotransfected reporter gene expression, indicating an inhibitory function of both molecules. Thus, SARA and Hgs are regulators of TGF-beta1 susceptibility in T cells and integrate regulatory signals into the influence of TGF-beta1-mediated suppression of human T cells. PMID- 12554699 TI - The prodomain of interleukin 1alpha interacts with elements of the RNA processing apparatus and induces apoptosis in malignant cells. AB - Interleukin 1alpha (IL-1alpha), a 33 kDa precursor, is cleaved releasing the 17 kDa carboxyl-terminal cytokine IL-1alpha to which all of the biological properties of IL-1alpha have been attributed. We investigated the potential independent properties of the remaining 16 kDa IL-1alpha amino-terminal propiece by expression in human tumor and primary human cell lines. The IL-1alpha propiece produced apoptosis in malignant but not normal cell lines. A minimal fragment comprised of amino acids 55-108 was required for apoptosis. Deletion and mutation studies identified an extended nuclear localization sequence required for nuclear localization, induction of apoptosis and concentration of the IL-1alpha propiece in interchromatin granule clusters, concentrations of proteins in the RNA splicing and processing pathways. The IL-1alpha propiece interacted with five known components of the RNA splicing/processing pathway, suggesting that the mechanism of action may involve changes in RNA splicing or processing. Expression of the IL-1alpha propiece caused a shift in the ratio of Bcl-Xl/Bcl-Xs toward the apoptotic direction. Our findings indicate that the IL-1alpha propiece induces apoptosis in a range of tumor cells and likely operates through a mechanism involving the RNA processing apparatus and the alternate splicing of apoptosis regulatory proteins. PMID- 12554700 TI - The role of readthrough acetylcholinesterase in the pathophysiology of myasthenia gravis. AB - Alternative splicing induces, under abnormal cholinergic neurotransmission, overproduction of the rare "readthrough" acetylcholinesterase variant AChE-R. We explored the pathophysiological relevance of this phenomenon in patients with myasthenia gravis (MG) and rats with experimental autoimmune MG (EAMG), neuromuscular junction diseases with depleted acetylcholine receptors. In MG and EAMG, we detected serum AChE-R accumulation. In EAMG, we alleviated electromyographic abnormalities by nanomolar doses of EN101, an antisense oligonucleotide that selectively lowers AChE-R in blood and muscle yet leaves unaffected the synaptic variant AChE-S. Whereas animals treated with placebo or conventional anticholinesterases continued to deteriorate, a 4 wk daily oral administration of EN101 improved survival, neuromuscular strength and clinical status in moribund EAMG rats. The efficacy of targeting only one AChE splicing variant highlights potential advantages of mRNA-targeted therapeutics for chronic cholinergic malfunctioning. PMID- 12554701 TI - Rho kinase and matrix metalloproteinase inhibitors cooperate to inhibit angiogenesis and growth of human prostate cancer xenotransplants. AB - The purpose of this study was to determine the effects of inhibitors of Rho kinase (ROK) and matrix metalloproteinases (MMPs) on angiogenesis and tumor growth and to evaluate ROK activity in human prostate cancer PC3 cells and endothelial cells (HUVECs). Vacuolation by endothelial cells and lumen formation, the earliest detectable stages of angiogenesis, were inhibited by the ROK inhibitor Wf-536. Combining Wf-536 with the MMP inhibitor Marimastat greatly enhanced in vitro inhibition of endothelial vacuolation, lumen and cord formation, and VEGF- and HGF-stimulated endothelial sprout formation from aorta. Inhibition of sprout formation by the two inhibitors was synergistic. Both agents inhibited migration of HUVECs. The regulatory subunit (MYPT1) of the myosin phosphatase was phosphorylated in PC3 cells and HUVECs, and phosphorylation of MYPT1 and the myosin regulatory light chain was reduced by Wf-536, providing direct evidence of ROK activity. Early treatment of immuno-incompetent mice bearing xenotransplants of PC3 cells with a combination of Wf-536 plus Marimastat with or without Paclitaxel, significantly inhibited tumor growth, prevented tumor growth escape after discontinuation of Paclitaxel, and increased survival. PMID- 12554702 TI - p11 regulates extracellular plasmin production and invasiveness of HT1080 fibrosarcoma cells. AB - The defining characteristic of a tumor cell is its ability to escape the constraints imposed by neighboring cells, invade the surrounding tissue, and metastasize to distant sites. This invasive property of tumor cells is dependent on activation of proteases at the cell surface. Most cancer cells secrete the urokinase-type plasminogen activator, which converts cell-bound plasminogen to plasmin. Here we address the issue of whether the plasminogen binding protein, p11, plays a significant role in this process. Transfection of human HT1080 fibrosarcoma cells with the human p11 gene in the antisense orientation resulted in a loss of p11 protein from the cell surface and concomitant decreases in cellular plasmin production, ECM degradation, and cellular invasiveness. The transfected cells demonstrated reduced development of lung metastatic foci in SCID mice. In contrast, HT1080 cells transfected with the p11 gene in the sense orientation displayed increased cell surface p11 protein and concomitant increases in cellular plasmin production, as well as enhanced ECM degradation and enhanced cellular invasiveness. The p11 overexpressing cells showed enhanced development of lung metastatic foci. These data establish that changes in the extracellular expression of the plasminogen receptor protein, p11, dramatically affect tumor cell-mediated pericellular proteolysis. PMID- 12554703 TI - Plant culture: thirteen seasonal pieces. February--constructing a corymb. AB - According to Robert Graves' mythological interpretation of the seasonal cycle, February is associated with the rowan tree. Corymb is the systematic name for the rowan inflorescence that produces the distinctive group of orange berries. The corymb, a gently domed cluster, is related to a panicle and an anthela, differing from them in the extent of upgrowth of the inflorescence branches. Soon, molecular biology will provide a gene-driven description of each inflorescence structure. This advance illustrates the progress of science: previous issues of inflorescence classification are not resolved, but a new system, reflecting the availability of new technology, is established. The artist and poet William Blake remarked that it is better to invent a system, than be enslaved by that of another. He also railed against science for its improper restriction to the imagination. PMID- 12554704 TI - CO2 concentrating mechanisms in cyanobacteria: molecular components, their diversity and evolution. AB - Cyanobacteria have evolved an extremely effective single-cell CO(2) concentrating mechanism (CCM). Recent molecular, biochemical and physiological studies have significantly extended current knowledge about the genes and protein components of this system and how they operate to elevate CO(2) around Rubisco during photosynthesis. The CCM components include at least four modes of active inorganic carbon uptake, including two bicarbonate transporters and two CO(2) uptake systems associated with the operation of specialized NDH-1 complexes. All these uptake systems serve to accumulate HCO(3)(-) in the cytosol of the cell, which is subsequently used by the Rubisco-containing carboxysome protein micro compartment within the cell to elevate CO(2) around Rubisco. A specialized carbonic anhydrase is also generally present in this compartment. The recent availability of at least nine cyanobacterial genomes has made it possible to begin to undertake comparative genomics of the CCM in cyanobacteria. Analyses have revealed a number of surprising findings. Firstly, cyanobacteria have evolved two types of carboxysomes, correlated with the form of Rubisco present (Form 1A and 1B). Secondly, the two HCO(3)(-) and CO(2) transport systems are distributed variably, with some cyanobacteria (Prochlorococcus marinus species) appearing to lack CO(2) uptake systems entirely. Finally, there are multiple carbonic anhydrases in many cyanobacteria, but, surprisingly, several cyanobacterial genomes appear to lack any identifiable CA genes. A pathway for the evolution of CCM components is suggested. PMID- 12554705 TI - Phosphorylation of a member of the MBF1 transcriptional co-activator family, StMBF1, is stimulated in potato cell suspensions upon fungal elicitor challenge. AB - StMBF1 (Solanum tuberosum multiprotein bridging factor 1) is a plant member of the MBF1 family of transcriptional co-activators. Previously, it has been described as being up-regulated at the transcriptional level by fungal and abiotic stress. To understand whether StMBF1 is also regulated at the post translational level, in vitro as well as in vivo phosphorylation assays were performed. StMBF1 is phosphorylated under both experimental conditions and [(32)P] incorporation into StMBF1 increases after treatment of potato cells with hyphal cell wall components (HWC) derived from Phytophthora infestans. The StMBF1 phosphorylating activity is strongly inhibited by the calcium-chelator EGTA and partially inhibited by calmodulin antagonists. Using bacterial purified StMBF1 as a substrate, a 57 kDa calcium-dependent protein kinase (p57) that is able to phosphorylate StMBF1 was detected. The StMBF1 kinase activity of p57 was higher in elicited than in non-treated cells. The role of the elicitor-dependent phosphorylation of StMBF1 is discussed. PMID- 12554706 TI - Cloning and characterization of two ripening-related strawberry (Fragaria x ananassa cv. Chandler) pectate lyase genes. AB - Two genomic clones corresponding to putative pectate lyase genes (plA and plB) were isolated and characterized in strawberry (Fragaria x ananassa cv. Chandler). The corresponding ORFs for the plA and plB genes revealed deduced proteins of 451 and 439 amino acids, respectively, that differ from that of the previously isolated strawberry plC gene. Southern blot analysis has shown that while the plB gene is a single copy gene, the plA gene is probably encoded by a small multigene family. By using specific probes corresponding to the untranslated 3' terminal region of the pl genes, and QRT-PCR methodology, the spatio-temporal expression pattern of both strawberry pl genes have been compared with that of the plC gene. The three transcripts were specifically expressed only in fruit and mainly during the ripening stages. Moreover, the expression of the plA and plB genes was induced in green de-achened fruit, but this increase was reduced by the external application of auxins as was the expression of plC. The expression of both pl genes was also strongly reduced in harvested fruit kept in controlled atmosphere (CA) containing high CO(2) levels. Immunolocalization studies using antibodies raised against the strawberry PL proteins placed the proteins in the cell wall of parenchymatic cells of the fruit receptacle. The role of pl genes in cell-wall disassembly and fruit ripening softening is discussed. PMID- 12554707 TI - Multiple signalling pathways mediate fungal elicitor-induced beta-thujaplicin biosynthesis in Cupressus lusitanica cell cultures. AB - The biosynthesis of a phytoalexin, beta-thujaplicin, in Cupressus lusitanica cell cultures can be stimulated by a yeast elicitor, H(2)O(2), or methyl jasmonate. Lipoxygenase activity was also stimulated by these treatments, suggesting that the oxidative burst and jasmonate pathway may mediate the elicitor-induced accumulation of beta-thujaplicin. The elicitor signalling pathway involved in beta-thujaplicin induction was further investigated using pharmacological and biochemical approaches. Treatment of the cells with calcium ionophore A23187 alone stimulated the production of beta-thujaplicin. A23187 also enhanced the elicitor-induced production of beta-thujaplicin. EGTA, LaCl(3), and verapamil pretreatments partially blocked A23187- or yeast elicitor-induced accumulation of beta-thujaplicin. These results suggest that Ca(2+) influx is required for elicitor-induced production of beta-thujaplicin. Treatment of cell cultures with mastoparan, melittin or cholera toxin alone or in combination with the elicitor stimulated the production of beta-thujaplicin or enhanced the elicitor-induced production of beta-thujaplicin. The G-protein inhibitor suramin inhibited the elicitor-induced production of beta-thujaplicin, suggesting that receptor-coupled G-proteins are likely to be involved in the elicitor-induced biosynthesis of beta thujaplicin. Indeed, both GTP-binding activity and GTPase activity of the plasma membrane were stimulated by elicitor, and suramin and cholera toxin affected G protein activities. In addition, all inhibitors of G-proteins and Ca(2+) flux suppressed elicitor-induced increases in lipoxygenase activity whereas activators of G-proteins and the Ca(2+) signalling pathway increased lipoxygenase activity. These observations suggest that Ca(2+) and G-proteins may mediate elicitor signals to the jasmonate pathway, and the jasmonate signalling pathway may then lead to the production of beta-thujaplicin. PMID- 12554708 TI - Potassium activities in cell compartments of salt-grown barley leaves. AB - Triple-barrelled microelectrodes measuring K(+) activity (a(K)), pH and membrane potential were used to make quantitative measurements of vacuolar and cytosolic a(K) in epidermal and mesophyll cells of barley plants grown in nutrient solution with 0 or 200 mM added NaCl. Measurements of a(K) were assigned to the cytosol or vacuole based on the pH measured. In epidermal cells, the salt treatment decreased a(K) in the vacuole from 224 to 47 mM and in the cytosol from 68 to 15 mM. In contrast, the equivalent changes in the mesophyll were from 235 to 150 mM (vacuole) and 79 to 64 mM (cytosol). Thus mechanisms exist to ameliorate the effects of salt on a(K) in compartments of mesophyll cells, presumably to minimize any deleterious consequences for photosynthesis. Thermodynamic calculations showed that K(+) is actively transported into the vacuole of both epidermal and mesophyll cells of salinized and non- salinized plants. Comparison of the values of a(K) in K(+)-replete, non-salinized leaf cells with those previously measured in root cells of plants grown under comparable conditions indicates that cytosolic a(K) is similar in cells of both organs, but vacuolar a(K) in leaf cells is approximately twice that in roots. This suggests differences in the regulation of vacuolar a(K), but not cytosolic a(K), in leaf and root cells. PMID- 12554709 TI - Potassium-selective channel in the red beet vacuolar membrane. AB - In higher plants the vacuolar K(+)-selective (VK) channel was identified solely in guard cells. This patch-clamp study describes a 40 pS homologue of the VK channel in Beta vulgaris taproot vacuoles. This voltage-independent channel is activated by submicromolar Ca(2+), and is ideally selective for K(+) over Cl(-) and Na(+). PMID- 12554710 TI - Arabidopsis mutants sensitive to gamma radiation include the homologue of the human repair gene ERCC1. AB - Mutants sensitive to ionizing radiation in yeast and mammals include an assortment of DNA repair genes. The majority of these DNA repair genes are involved in the repair of DNA double-strand breaks. In this study a forward genetic screen is used to identify gamma-sensitive mutants of Arabidopsis thaliana. The gamma-plantlet screen used here also reveals two general mutant classes based on size of cotyledons and hypocotyls. One of the mutants discovered is a homologue of the mammalian nucleotide excision repair gene ERCC1. PMID- 12554711 TI - Knotted1-like genes are strongly expressed in differentiated cell types in sunflower. AB - Knotted1-like genes constitute a family of genes whose products are transcription factors involved in several aspects of plant development. In most species, these genes are expressed primarily in meristematic cells and are switched off as leaves develop. In this paper, the expression patterns of three kn1-like genes from sunflower (Helianthus annuus L.) are described. Northern blot experiments indicated that these genes are expressed at different levels in several organs of the plant, including flowers, leaves, stems, roots, and embryos. Notably, one of these genes, named HAKN1, was also highly expressed in leaves and roots. Using in situ hybridization, expression was detected in parenchymatic cells from leaf veins, petiole and lamina, and also in stem and root. Enhanced expression in phloem was also evident in both leaves and stem. Another, HAKN2, showed preferential expression in stem, specifically in fascicular and interfascicular cambium and phloem. In flowers, both genes are expressed throughout inflorescence and floral meristems and in developing organ primordia. Strong expression of HAKN1 in developing involucral bracts was also observed. The results show the existence of some differences in expression patterns of kn1-like genes in sunflower with respect to other plants. It is proposed that cell- and species specific factors are involved in determining the developmental responses of plant cells to the expression of kn1-like genes. PMID- 12554712 TI - In vitro distribution and characterization of membrane-associated PLD and PI-PLC in Brassica napus. AB - Two types of phospholipid degrading enzyme, phospholipase D (PLD; EC 3.1.4.4) and phosphatidyl- inositol-specific phospholipase C (PIP(2)-PLC; PI-PLC 3.1.4.11) were studied during the development of seeds and plants of Brassica napus. PLD exhibits two types of activity; polyphosphoinositide-requiring (PIP(2)-dependent PLD) and polyphosphoinositide-independent requiring millimolar concentrations of calcium (PLDalpha). Significantly different patterns of activity profiles were found for soluble and membrane-associated forms of all three enzymes within both processes. Membrane-associated PIP(2)-dependent PLD activity shows the opposite trend when compared to PLDalpha, while the highest PI-PLC activity appears in the same stages of development of seeds and plants as for PLDalpha. In subcellular fractions of hypocotyls of young plants, phospholipases were localized predominantly on plasma membranes. The biochemical characteristics (Ca(2+), pH) of all three enzymes associated with plasma membrane vesicles, isolated by partitioning in an aqueous dextran- polyethylene glycol two-phase system, are also described. Direct interaction of PLDalpha with G-proteins under in vitro conditions was not confirmed. PMID- 12554713 TI - Genes encoding two essential DNA replication activation proteins, Cdc6 and Mcm3, exhibit very different patterns of expression in the tobacco BY-2 cell cycle. AB - Very little is known about the expression patterns in plants of genes that encode proteins involved in the initiation of DNA replication. Partial cDNA sequences that encode Cdc6 and Mcm3 in tobacco have been isolated. The sequences were used as probes in northern blots which suggested that, in the cell cycle of synchronized tobacco BY-2 cells, expression of CDC6 is confined to late G(1) and S-phase whereas the expression of MCM3 is not confined to any particular cell cycle phase. These data were confirmed and extended by real-time PCR measurements of mRNA abundance through the cell cycle. CDC6 exhibits a very clear peak of expression in S-phase whereas MCM3, expressed at a much lower level than CDC6, is not cell-cycle-regulated. These patterns of cell cycle gene expression resemble those found in the fission yeast Schizosaccharomyces pombe rather than those in budding yeast or mammalian cells. PMID- 12554714 TI - Dramatic difference in the responses of phosphoenolpyruvate carboxylase to temperature in leaves of C3 and C4 plants. AB - Temperature caused phenomenal modulation of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) in leaf discs of Amaranthus hypochondriacus (NAD-ME type C(4) species), compared to the pattern in Pisum sativum (a C(3) plant). The optimal incubation temperature for PEPC in A. hypochondriacus (C(4)) was 45 degrees C compared to 30 degrees C in P. sativum (C(3)). A. hypochondriacus (C(4)) lost nearly 70% of PEPC activity on exposure to a low temperature of 15 degrees C, compared to only about a 35% loss in the case of P. sativum (C(3)). Thus, the C(4) enzyme was less sensitive to supra-optimal temperature and more sensitive to sub-optimal temperature than that of the C(3) species. As the temperature was raised from 15 degrees C to 50 degrees C, there was a sharp decrease in malate sensitivity of PEPC. The extent of such a decrease in C(4) plants (45%) was more than that in C(3) species (30%). The maintenance of high enzyme activity at warm temperatures, together with a sharp decrease in the malate sensitivity of PEPC was also noticed in other C(4) plants. The temperature-induced changes in PEPC of both A. hypochondriacus (C(4)) and P. sativum (C(3)) were reversible to a large extent. There was no difference in the extent of phosphorylation of PEPC in leaves of A. hypochondriacus on exposure to varying temperatures, unlike the marked increase in the phosphorylation of enzyme on illumination of the leaves. These results demonstrate that (i) there are marked differences in the temperature sensitivity of PEPC in C(3) and C(4) plants, (ii) the temperature induced changes are reversible, and (iii) these changes are not related to the phosphorylation state of the enzyme. The inclusion of PEG-6000, during the assay, dampened the modulation by temperature of malate sensitivity of PEPC in A. hypochondriacus. It is suggested that the variation in temperature may cause significant conformational changes in C(4)-PEPC. PMID- 12554715 TI - Subcellular localization of ADPglucose pyrophosphorylase in developing wheat endosperm and analysis of the properties of a plastidial isoform. AB - The intracellular location of ADPglucose pyrophosphorylase (AGPase) in wheat during endosperm development was investigated by analysis of the recovery of marker enzymes from amyloplast preparations. Amyloplast preparations contained 20 28% of the total endosperm activity of two plastidial marker enzymes and less than 0.8% of the total endosperm activity of two cytosolic marker enzymes. Amylo plasts prepared at various stages of development, from 8-30 d post anthesis, contained between 2% and 10% of the total AGPase activity; this implies that between 7% and 40% of the AGPase in wheat endosperm is plastidial during this period of development. Two proteins were recognized by antibodies to both the large and small subunits of wheat AGPase. The larger of the two AGPases was the major form of the enzyme in whole cell extracts, and the smaller, less abundant, form of AGPase was enriched in plastid preparations. The results are consistent with data from other graminaceous endosperms, suggesting that there are distinct plastidial and cytosolic isoforms of AGPase composed of different subunits. The plastidial isoform of AGPase from wheat endosperm is relatively insensitive to the allosteric regulators 3-phosphoglycerate and inorganic orthophos phate compared with plastidial AGPase from other species. Amyloplast AGPase showed no sensitivity to physiological concentrations of inorganic orthophosphate. 15 mM 3 phosphoglycerate caused no stimulation of the pyrophosphorolytic reaction, and only 2-fold stimulation of the ADPglucose synthesizing reaction. PMID- 12554716 TI - Characterization of Pinalate, a novel Citrus sinensis mutant with a fruit specific alteration that results in yellow pigmentation and decreased ABA content. AB - The characterization of a novel mutant, named Pinalate, derived from the orange (Citrus sinensis L. Osbeck) Navelate, which produces distinctive yellow fruits instead of the typical bright orange colouration, is reported. The carotenoid content and composition, and ABA content in leaf and flavedo tissue (coloured part of the skin) of fruits at different developmental and maturation stages were analysed. No important differences in leaf carotenoid pattern of both phenotypes were found. However, an unusual accumulation of linear carotenes (phytoene, phytofluene and zeta- carotene) was detected in the flavedo of Pinalate. As fruit maturation progressed, the flavedo of mutant fruit accumulated high amounts of these carotenes and the proportion of cyclic and oxygenated carotenoids was substantially lower than in the parental line. Full-coloured fruit of Pinalate contained about 44% phytoene, 21% phytofluene, 25% zeta-carotene, and 10% of xanthophylls, whereas, in Navelate, 98% of total carotenoids were xanthophylls and apocarotenoids. The ABA content in the flavedo of Pinalate mature fruit was 3 6 times lower than in the corresponding tissue of Navelate, while no differences were found in leaves. Other maturation processes were not affected in Pinalate fruit. Taken together, the results indicate that Pinalate is a fruit-specific alteration defective in zeta-carotene desaturase or in zeta-carotene desaturase associated factors. Possible mechanisms responsible for the Pinalate phenotype are discussed. Because of the abnormal fruit-specific carotenoid complement and ABA deficiency, Pinalate may constitute an excellent system for the study of carotenogenesis in Citrus and the involvement of ABA in fruit maturation and stress responses. PMID- 12554717 TI - Antisense SNF1-related (SnRK1) protein kinase gene represses transient activity of an alpha-amylase (alpha-Amy2) gene promoter in cultured wheat embryos. AB - A DNA fragment corresponding to part of an SNF1 (sucrose non-fermenting-1) related protein kinase (SnRK1) transcript was amplified by a polymerase chain reaction (PCR) from a wheat (Triticum aestivum) endosperm cDNA library. It was used to construct a chimaeric gene, pUasSnRKN, comprising a ubiquitin promoter, the SnRK1 PCR product in the antisense orientation and the nopaline synthase (Nos) gene terminator. This construct was used in transient gene expression experiments in cultured wheat embryos together with a series of reporter gene constructs. These included the wheat alpha amylase gene alpha-Amy2 promoter with UidA (Gus) coding region (palpha2GT), rice actin promoter with Gus (pActIDGus), ubiquitin promoter with Gus (pAHC25) and actin promoter with green fluorescent protein (GFP) gene (pAct1Is-GFP1). All of the reporter genes were found to be active when bombarded into scutellae isolated from immature grains at 25 d post anthesis and incubated on MS medium for 24 h prior to bombardment. However, co bombardment of palpha2GT with equimolar amounts of pUasSnRKN resulted in no detectable Gus activity, indicating that the antisense SnRK1 construct repressed the alpha-Amy2 promoter. Co-bombardment with pUasSnRKN had no effect on the activity of the other promoters used in the study. A triple bombardment with palpha2GT, pAct1Is-GFP-1 and pUasSnRKN resulted in clear green fluorescence, indicating that the bombarded cells were still viable, but no Gus activity. RT PCR analysis showed clearly that the antisense SnRK1 gene was expressing. Northern and RT-PCR analyses confirmed that SnRK1 and both alpha-amylase genes, alpha-Amy1 and alpha-Amy2, are expressed in cultured wheat embryos harvested from grain 25 d post-anthesis. Expression of alpha-Amy1 and alpha-Amy2 was up regulated by sugar starvation. PMID- 12554718 TI - Heterogeneity of sucrose synthase genes in bean (Phaseolus vulgaris L.): evidence for a nodule-enhanced sucrose synthase gene. AB - Sucrose synthase (SS), the key sucrose hydrolytic enzyme (EC 2.4.1.13), plays an important role in N(2)-fixing nodule metabolism. It has also been proposed that N(2) fixation in soybean nodules could be mediated by the potential to metabolize sucrose. The isolation and characterization of a nodule-enhanced SS full-length cDNA clone from the bean Phaseolus vulgaris is reported here. Southern blot analysis indicated that there are at least two SS genes in beans. Using a 3' specific probe from this SS cDNA clone, it was possible to identify a nodule enhanced SS gene (PvSSn), which is expressed almost exclusively in nodules. A second gene (PvSS), which is expressed in all tissues tested, was detected using a coding region probe. Nodule-enhanced PvSSn transcript levels, but not the enzyme activity or protein amount, is reduced during nodule development. These data indicated that this reduction could be due to a limitation on the carbon availability in the nodule. PvSSn expression is reduced in the asparagine-treated nodules. By contrast, PvSSn transcript levels in nodules increased in the presence of glutamine, allantoin and allopurinol. This result suggests a relationship between ureide transport and SS regulation and could help in understanding why the ureide transport mechanism is activated during nitrogen fixation in bean. PMID- 12554719 TI - UV-induced blue-green and far-red fluorescence along wheat leaves: a potential signature of leaf ageing. AB - Under UV-excitation, leaves emit red (RF) and far-red (FRF) fluorescence from chlorophyll and blue-green fluorescence (BGF) from hydroxycinnamic acids. In this study, the aim was to develop a fluorescence signature of wheat leaf ageing after the emergence of the lamina. FRF and BGF were examined in the first three leaves of 2-week-old wheat plants. It was investigated how FRF and BGF vary as leaf and tissue aged by spectroscopic measurements, time-resolved BGF analysis and microscopic imaging of the leaf surface. It was found that FRF decreased with leaf and tissue ageing because of an accumulation of UV-absorbers in the epidermis. BGF also decreased, but without changes either in the shape of excitation and emission spectra or in the fluorescence lifetime. So, BGF emanated from the leaf surface, without changes in fluorophore composition during leaf ageing. The shape of the BGF spectrum indicates that ferulic acid bound to the cell wall is the main blue-green fluorophore. The effects of pH and solvents on BGF from intact leaves and ferulic acid in solution were similar, confirming the hydroxycinnamic acid origin of BGF. UV-fluorescence microscopic imaging of the surface of intact leaves showed that different epidermis cell types and sclerenchyma bands emitted BGF. The decreasing gradient of BGF from the base to the apex of the lamina could be related to the decrease in the surface of the fluorescent sclerenchyma bands. The significance of FRF and BGF as potential signatures of wheat lamina growth are discussed. PMID- 12554720 TI - Ethylene is required for both the initiation and progression of softening in pear (Pyrus communis L.) fruit. AB - In order to investigate the physiological role of ethylene in the initiation and subsequent progression of softening, pear fruit were treated with propylene, an analogue of ethylene or 1-methylcyclopropene (1-MCP), a gaseous inhibitor of ethylene action at the preclimacteric or ripening stages. The propylene treatment at the pre-ripe stage stimulated ethylene production and flesh softening while the 1-MCP treatment at the same stage markedly retarded the initiation of the ripening-related events. Moreover, 1-MCP treatment after the initiation of ripening markedly suppressed the subsequent flesh softening and ethylene production. These results clearly indicate that ethylene is not merely a by product, but plays a crucial role in both the initiation and maintenance of regulating the softening process during ripening. The observations also suggest that ethylene in ripening is regulated entirely in an autocatalytic manner. The mRNA accumulation of pear polygalacturonases (PG) genes, PC-PG1 and PC-PG2, was in parallel with the pattern of fruit softening in both propylene and 1-MCP treatments. However, the expression pattern of pear endo-1,4-beta-D-glucanases (EGase) genes, PC-EG1 and PC-EG2, was not affected in both treatments. The results suggest that ethylene is required for PGs expression even in the late ripening stage, but not for EGases. PMID- 12554721 TI - AKT1 and TRH1 are required during root hair elongation in Arabidopsis. AB - TRH1 is a member of the AtKT/AtKUP/AtHAK family of potassium carriers that is required for root hair elongation and AKT1 is an inward rectifying potassium channel expressed in the root epidermis, endodermis and cortex of Arabidopsis thaliana. Plants homozygous for the trh1-1 mutation form short root hairs. The Trh1(-) phenotype cannot be suppressed by growing plants homozygous for the trh1 1 mutation in the presence of high external KCl concentration. This indicates an absolute requirement for TRH1 in root hair tip growth. Plants homozygous for the akt1-1 mutation develop longer root hairs than the wild type when grown in 0 mM external potassium, but develop shorter hairs than the wild type when grown in higher concentrations [>10 mM] of potassium. These data indicate that both TRH1 and AKT1 are active in the root hair over a wide range of external potassium concentrations, but suggest they have different functions in the growing hair cell. PMID- 12554722 TI - Characterization of nitrogen relationships between Sorghum bicolor and the root hemiparasitic angiosperm Striga hermonthica (Del.) Benth. using K15 NO3 as isotopic tracer. AB - The role of the host in the nitrogen nutrition of Striga hermonthica (Del.) Benth. (Scrophulariaceae) parasitic on Sorghum bicolor cv. SH4 Arval has been investigated using (15)N-nitrate as the tracer. It is shown that, when nitrate is absorbed only by the roots of the host plant, a rapid transfer of nitrogen to the parasite can be detected. The xylem sap of S. hermonthica contained approximately equal amounts of nitrate and amino acids, mostly glutamine and asparagine. Infection altered the free amino acid profile of the host tissues, leading notably to a large increase in asparagine and a decrease in glutamine. The haustoria of S. hermonthica, although rich in nitrate, showed a low concentration of free amino acids, particularly lacking in asparagine and glutamine. The roots of S. hermonthica, in contrast, were rich in both asparagine and glutamine while, in the shoots, asparagine constituted 80% of the total FAA pool. Asparagine was also found to be the primary (15)N-enriched amino acid in the shoots of S. hermonthica while, interestingly, it was glutamate that was most strongly enriched in the roots. It is concluded that nitrogen nutrition in S. hermonthica is based on a supply of both nitrate and amino acids from the host. This implies a non-specific transfer in the transpiration stream. Nitrate reduction probably occurs mainly in the leaves of the parasite. Assimilation also occurs in S. hermonthica and excess nitrogen is stored as the non-toxic nitrogen-rich compound, asparagine. This specific trait of nitrogen metabolism of the parasite is discussed in relation to the effect of nitrogen fertilization on reducing infestation. PMID- 12554723 TI - Mapping of QTL associated with nitrogen storage and remobilization in barley (Hordeum vulgare L.) leaves. AB - Nitrogen uptake and metabolism are central for vegetative and reproductive plant growth. This is reflected by the fact that nitrogen can be remobilized and reused within a plant, and this process is crucial for yield in most annual crops. A population of 146 recombinant inbred barley lines (F(8) and F(9) plants, grown in 2000 and 2001), derived from a cross between two varieties differing markedly in grain protein concentration, was used to compare the location of QTL associated with nitrogen uptake, storage and remobilization in flag leaves relative to QTL controlling developmental parameters and grain protein accumulation. Overlaps of support intervals for such QTL were found on several chromosomes, with chromosomes 3 and 6 being especially important. For QTL on these chromosomes, alleles associated with inefficient N remobilization were associated with depressed yield and higher levels of total or soluble organic nitrogen during grain filling and vice versa; therefore, genes directly involved in N recycling or genes regulating N recycling may be located on these chromosomes. Interestingly, the most prominent QTL for grain protein concentration (on chromosome 6) did not co-localize with QTL for nitrogen remobilization. However, QTL peaks for nitrate and soluble organic nitrogen were detected at this locus for plants grown in 2001 (but not in 2000). For these, alleles associated with low grain protein concentration were associated with higher soluble nitrogen levels in leaves during grain filling; therefore, gene(s) found at this locus might influence the nitrogen sink strength of developing barley grains. PMID- 12554724 TI - Electrophysiological responses of maize roots to low water potentials: relationship to growth and ABA accumulation. AB - The maintenance of root elongation is an important adaptive response to low water potentials (psi(w)), but little is known about its regulation. An important component may be changes in root cell electrophysiology, which both signal and maintain growth maintenance processes. As a first test of this hypothesis, membrane potentials (E(m)) were measured within the cell elongation zone of maize (Zea mays L.) primary roots. Seedlings were grown in oxygenated solution culture, and low psi(w) was imposed by the gradual addition of polyethylene glycol. Cells hyperpolarized approximately 25 mV in response to low psi(w), and after 48 h resting potentials remained significantly hyperpolarized at psi(w) lower than 0.3 MPa compared with roots at high psi(w). Inhibitor experiments showed that the hyperpolarization was dependent on plasma membrane H(+)-ATPase activity. Previous work showed that accumulation of abscisic acid (ABA) is required for the maintenance of maize primary root elongation at low psi(w). To determine if the mechanism of action of ABA involves changes in root electrophysiology, E(m) measurements were made during long-term exposure to low psi(w). Steady-state resting E(m) were measured in regions in which maintenance of cell elongation was dependent on ABA accumulation (2-3 mm from the apex), or in which elongation was inhibited regardless of ABA status (6-8 mm from the apex). E(m) was substantially more negative in ABA-deficient roots specifically in the 2-3 mm region. The results suggest that set-points for ion homeostasis shifted in association with the maintenance of root cell elongation at low psi(w), and that ABA accumulation plays a role in regulating the ion transport processes involved in this response. PMID- 12554725 TI - Chloride absorption in salt-sensitive Carrizo citrange and salt-tolerant Cleopatra mandarin citrus rootstocks is linked to water use. AB - In this work, seedlings of two citrus rootstocks, the salt-tolerant Cleopatra mandarin (Citrus reshni Hort. ex Tan.) and the salt-sensitive Carrizo citrange (Citrus sinensis [L.] Osb. x Poncirus trifoliata [L.] Raf.) were used to study the relationship between chloride and water uptake. The results indicated that net chloride uptake rates in both genotypes were alike and decreased linearly with the time of salinity exposure, although they were more rapidly reduced in the tolerant genotype. In each rootstock, chloride uptake rates paralleled the decreases in transpiration rates. When transpiration was modified, concomitant changes in leaf Cl(-) concentrations were observed. There was a high positive correlation between total chloride content per plant and total water absorbed. In addition, the data indicate that the tolerant genotype "excluded" more chloride, i.e. it absorbed lower amounts of chloride per volume of water. Cleopatra also possessed a less efficient root system for water uptake and a higher shoot-to root ratio. The results show that, overall, chloride absorption is linked to water use and that further tolerance in Cleopatra is mostly conferred by superior root resistance to Cl(-) uptake. Therefore, it is proposed that chloride absorption and, hence, salt tolerance in citrus depends to a great extent upon water use. PMID- 12554726 TI - Transgenic loblolly pine (Pinus taeda L.) plants expressing a modified delta endotoxin gene of Bacillus thuringiensis with enhanced resistance to Dendrolimus punctatus Walker and Crypyothelea formosicola Staud. AB - A synthetic version of the CRY1Ac gene of Bacillus thuringiensis has been used for the transformation of loblolly pine (Pinus taeda L.) using particle bombardment. Mature zygotic embryos were used to be bombarded and to generate organogenic callus and transgenic regenerated plants. Expression vector pB48.215 DNA contained a synthetic Bacillus thuringiensis (B.t.) CRY1Ac coding sequence flanked by the double cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (NOS) terminator sequences, and the neomycin phosphotransferase II (NPTII) gene controlled by the promoter of the nopaline synthase gene was introduced into loblolly pine tissues by particle bombardment. The transformed tissues were proliferated and selected on media with kanamycin. Shoot regeneration was induced from the kanamycin-resistant calli, and transgenic plantlets were then produced. More than 60 transformed plants from independent transformation events were obtained for each loblolly pine genotype tested. The integration and expression of the introduced genes in the transgenic loblolly pine plants was confirmed by polymerase chain reactions (PCR) analysis, by Southern hybridization, by Northern blot analysis, and by Western blot analysis. Effective resistance of transgenic plants against Dendrolimus punctatus Walker and Crypyothelea formosicola Staud was verified in feeding bioassays with the insects. The transgenic plants recovered could represent a good opportunity to analyse the impact of genetic engineering of pine for sustainable resistance to pests using a B. thuringiensis insecticidal protein. This protocol enabled the routine transformation of loblolly pine plants that were previously difficult to transform. PMID- 12554727 TI - The effects of air flow and stem flexure on the mechanical and hydraulic properties of the stems of sunflowers Helianthus annuus L. AB - Many studies have shown that wind affects plant development, causing them to develop shorter and usually stronger stems. Many of these effects have been shown to be due to a response to mechanical flexing of the stem which is known as thigmomorphogenesis. However, it is not known how wind affects the hydraulic properties of stems, nor have the effects of air flow past leaves been examined in isolation from mechanical flexing. This study, therefore, used a factorial experiment to distinguish between the effects of stem flexing and air flow, and examined the morphology, hydraulics and mechanics of developing sunflowers Helianthus annuus. It was found that flexure and air flow had opposite effects on several aspects of development; air flow increased plant height and length specific stem hydraulic conductivity, k(h), and reduced stem rigidity and strength, while flexing did the reverse. There was also a clear trade-off between hydraulic and mechanical capability: as one increased the other decreased. A plant's response to wind must, therefore, be a complex response to at least two different stimuli and this might help explain why it varies with species and environment. PMID- 12554728 TI - Salinity treatment shows no effects on photosystem II photochemistry, but increases the resistance of photosystem II to heat stress in halophyte Suaeda salsa. AB - Photosynthetic gas exchange, modulated chlorophyll fluorescence, rapid fluorescence induction kinetics, and the polyphasic fluorescence transients were used to evaluate PSII photochemistry in the halophyte Suaeda salsa exposed to a combination of high salinity (100-400 mM NaCl) and heat stress (35-47.5 degrees C, air temperature). CO(2) assimilation rate increased slightly with increasing salt concentration up to 300 mM NaCl and showed no decrease even at 400 mM NaCl. Salinity treatment showed neither effects on the maximal efficiency of PSII photochemistry (F(v)/F(m)), the rapid fluorescence induction kinetics, and the polyphasic fluorescence transients in dark-adapted leaves, nor effects on the efficiency of excitation energy capture by open PSII reaction centres (F(v)'/F(m)') and the actual PSII effciency (Phi(PSII)), photochemical quenching (q(P)), and non-photochemical quenching (q(N)) in light-adapted leaves. The results indicate that high salinity had no effects on PSII photochemistry either in a dark-adapted state or in a light-adapted state. With increasing temperature, CO(2) assimilation rate decreased significantly and no net CO(2) assimilation was observed at 47.5 degrees C. Salinity treatment had no effect on the response of CO(2) assimilation to high temperature when temperature was below 40 degrees C. At 45 degrees C, CO(2) assimilation rate in control plants decreased to zero, but the salt-adapted plants still maintained some CO(2) assimilation capacity. On the other hand, the responses of PSII photochemistry to heat stress was modified by salinity treatment. When temperature was above 35 degrees C, the declines in F(v)/F(m), Phi(PSII), F(v)'/F(m)', and q(P) were smaller in salt-adapted leaves compared to control leaves. This increased thermostability was independent of the degree of salinity, since no significant changes in the above-described fluorescence parameters were observed among the plants treated with different concentrations of NaCl. During heat stress, a very clear K step as a specific indicator of damage to the O(2)-evolving complex in the polyphasic fluorescence transients appeared in control plants, but did not get pronounced in salt-adapted plants. In addition, a greater increase in the ratio (F(i)-F(o))/(F(p)-F(o)) which is an expression of the proportion of the Q(B)-non-reducing PSII centres was observed in control plants rather than in salt-adapted plants. The results suggest that the increased thermostability of PSII seems to be associated with the increased resistance of the O(2)-evolving complex and the reaction centres of PSII to high temperature. PMID- 12554729 TI - Night-time conductance in C3 and C4 species: do plants lose water at night? AB - Significant night-time stomatal conductance and transpiration were found for 11 out of 17 species with a range of life histories (herbaceous annual, perennial grass, shrub, tree), photosynthetic pathways (C(3), C(4)), and habitats in the western United States. Across species and habitats, higher night-time conductance and transpiration were associated with higher daytime values. The prevalence, mechanisms and ecological implications of substantial night-time water loss deserve further investigation. PMID- 12554730 TI - Photosynthetic performance of an Arabidopsis mutant with elevated stomatal density (sdd1-1) under different light regimes. AB - In the Arabidopsis mutant sdd1-1, a point mutation in a single gene (SDD1) causes specific alterations in stomatal density and distribution. In comparison to the wild type (C24), abaxial surfaces of sdd1-1 rosette leaves have about 2.5-fold higher stomatal densities. This mutant was used to study the consequence of stomatal density on photosynthesis under various light regimes. The increased stomatal density in the mutant had no significant influence on the leaf CO(2) assimilation rate (A) under constant light conditions. Mutant and wild-type plants contained similar amounts of carbohydrates under these conditions. However, exposure of plants to increasing photon flux densities resulted in differences in gas exchange and the carbohydrate metabolism of the wild type and mutant. Increased stomatal densities in sdd1-1 enabled low-light-adapted plants to have 30% higher CO(2) assimilation rates compared to the wild type when exposed to high light intensities. After 2 d under high light conditions leaves of sdd1-1 accumulated 30% higher levels of starch and hexoses than wild-type plants. PMID- 12554732 TI - Posttranscriptional and posttranslational regulation of C/EBP delta in G0 growth arrested mammary epithelial cells. AB - Previous work from our laboratory demonstrated that CCAAT/enhancer-binding protein delta (C/EBP delta) functions in the initiation and maintenance of G(0) growth arrest in mouse mammary epithelial cells (MECs). In this report, we investigated the posttranscriptional and posttranslational regulation of C/EBP delta in G(0) growth-arrested mouse MECs. The results of transcriptional inhibitor studies demonstrated that the C/EBP delta mRNA exhibits a relatively short half-life in G(0) growth-arrested mouse MECs (t(1/2) approximately 35 min). In contrast, C/EBP delta mRNA has a longer half-life in G(0) growth-arrested mouse fibroblast cells (t(1/2) >100 min). Oligo/RNase H cleavage analysis and rapid amplification of cDNA ends-poly(A) test both confirmed the short C/EBP delta mRNA half-life observed in MECs and demonstrated that the C/EBP delta mRNA poly(A) tail is relatively short (approximately 100 nucleotides). In addition, the poly(A) tail length was not shortened during C/EBP delta mRNA degradation, which suggested a deadenylation-independent pathway. The C/EBP delta protein also exhibited a relatively short half-life in G(0) growth-arrested mouse MECs (t(1/2) approximately 120 min). The C/EBP delta protein was degraded in a ubiquitin dependent manner, primarily in the nucleus, during G(0) growth arrest. In conclusion, these studies indicated that the C/EBP delta mRNA and protein content are under tight regulation in G(0) growth-arrested mouse MECs, despite the general concept that G(0) growth arrest is associated with a decrease in cellular activity. PMID- 12554733 TI - Identification of a phosphothionate analogue of lysophosphatidic acid (LPA) as a selective agonist of the LPA3 receptor. AB - Lysophosphatidic acid (LPA) is a bioactive lysophospholipid mediator that acts through G protein-coupled receptors. Most cell lines in culture express one or more LPA receptors, making it difficult to assign a response to specific LPA receptors. Dissection of the signaling properties of LPA has been hampered by lack of LPA receptor subtype-specific agonists and antagonists. The present study characterizes an ester-linked thiophosphate derivative (1-oleoyl-2-O-methyl-rac glycerophosphothionate, OMPT) of LPA. OMPT is a functional LPA analogue with potent mitogenic activity in fibroblasts. In contrast to LPA, OMPT does not couple to the pheromone response through the LPA(1) receptor in yeast cells. OMPT induces intracellular calcium increases efficiently in LPA(3) receptor-expressing Sf9 cells but poorly in LPA(2) receptor-expressing cells. Guanosine 5'-O-(3 [(35)S]thio)triphosphate binding assays in mammalian cells showed that LPA exhibits agonistic activity on all three LPA receptor subtypes, whereas OMPT has a potent agonistic effect only on the LPA(3) receptor. In transiently transfected HEK293 cells, OMPT stimulates mitogen-activated protein kinases through the LPA(3) but not the LPA(1) or LPA(2) receptors. Furthermore, OMPT-induced intracellular calcium mobilization in mammalian cells is efficiently inhibited by the LPA(1)/LPA(3) receptor-selective antagonist VPC12249. These results establish that OMPT is an LPA(3)-selective agonist. OMPT binding to the LPA(3) receptor in mammalian cells is sufficient to elicit multiple responses, including activation of G proteins, calcium mobilization, and activation of mitogen-activated protein kinases. Thus OMPT offers a powerful probe for the dissection of LPA signaling events in complex mammalian systems. PMID- 12554734 TI - Arresting initiation of hepatitis C virus RNA synthesis using heterocyclic derivatives. AB - The hepatitis C virus (HCV) NS5B protein encodes an RNA-dependent RNA polymerase (RdRp), the primary catalytic enzyme of the HCV replicase complex. Recently, two benzo-1,2,4-thiadiazine compounds were shown to be potent, highly specific inhibitors of the genotype 1b HCV RdRp containing a carboxyl-terminal 21 residue truncation (delta21 HCV RdRp) (Dhanak, D., Duffy, K., Johnston, V. K., Lin Goerke, J., Darcy, M., Shaw, A. N. G. B., Silverman, C., Gates, A. T., Earnshaw, D. L., Casper, D. J., Kaura, A., Baker, A., Greenwood, C., Gutshall, L. L., Maley, D., DelVecchio, A., Macarron, R., Hofmann, G. A., Alnoah, Z., Cheng, H. Y., Chan, G., Khandekar, S., Keenan, R. M., and Sarisky, R. T. (2002) J. Biol. Chem. 277, 38322-38327). Compound 4 (C(21)H(21)N(3)O(4)S) reduces viral replication by virtue of its direct interaction with the viral polymerase rather than by nonspecific titration of nucleic acid template. In this study, we present several lines of evidence to demonstrate that this inhibitor interferes with the initiation step of RNA synthesis rather than acting as an elongation inhibitor. Inhibition of initial phosphodiester bond formation occurred regardless of whether replication was initiated by primer-dependent or de novo mechanisms. Filter binding studies using increasing concentrations of compound 4 did not interfere with the ability of delta21 HCV RdRp to interact with nucleic acid. Furthermore, varying the order of reagent addition in the primer extension assay showed no distinct differences in inhibition profile. Finally, surface plasmon resonance analyses provided evidence that a ternary complex is capable of forming between the RNA template, RdRp, and compound 4. Together, these data suggest that this heterocyclic agent interacts with the apoenzyme, as well as with the RNA bound form of delta21 HCV RdRp, and therefore does not directly interfere with the RdRp-RNA interaction to mediate inhibition. PMID- 12554735 TI - Inhibition of hepatitis C virus RNA replication by 2'-modified nucleoside analogs. AB - The RNA-dependent RNA polymerase (NS5B) of hepatitis C virus (HCV) is essential for the replication of viral RNA and thus constitutes a valid target for the chemotherapeutic intervention of HCV infection. In this report, we describe the identification of 2'-substituted nucleosides as inhibitors of HCV replication. The 5'-triphosphates of 2'-C-methyladenosine and 2'-O-methylcytidine are found to inhibit NS5B-catalyzed RNA synthesis in vitro, in a manner that is competitive with substrate nucleoside triphosphate. NS5B is able to incorporate either nucleotide analog into RNA as determined with gel-based incorporation assays but is impaired in its ability to extend the incorporated analog by addition of the next nucleotide. In a subgenomic replicon cell line, 2-C-methyladenosine and 2'-O methylcytidine inhibit HCV RNA replication. The 5'-triphosphates of both nucleosides are detected intracellularly following addition of the nucleosides to the media. However, significantly higher concentrations of 2'-C-methyladenosine triphosphate than 2'-O-methylcytidine triphosphate are detected, consistent with the greater potency of 2'-C-methyladenosine in the replicon assay, despite similar inhibition of NS5B by the triphosphates in the in vitro enzyme assays. Thus, the 2'-modifications of natural substrate nucleosides transform these molecules into potent inhibitors of HCV replication. PMID- 12554736 TI - Regulation of mammalian STE20-like kinase 2 (MST2) by protein phosphorylation/dephosphorylation and proteolysis. AB - Mammalian STE20-like kinase 2 (MST2), a member of the STE20-like kinase family, has been shown in previous studies to undergo proteolytic activation by caspase-3 during cell apoptosis. A few studies have also implicated protein phosphorylation reactions in MST2 regulation. In this study, we examined the mechanism of MST2 regulation with an emphasis on the relationship between caspase-3 cleavage and protein phosphorylation. Both the full-length MST2 and the caspase-3-truncated form of MST2 overexpressed in 293T cells exist in a phosphorylated state. On the other hand, the endogenous full-length MST2 from rat thymus or from proliferating cells is mainly unphosphorylated whereas the caspase-3-truncated endogenous MST2 from apoptotic cells is highly phosphorylated. Cell transfection studies using mutant MST2 constructs indicate that MST2 depends on the autophosphorylation of a unique threonine residue, Thr(180), for kinase activity. The autophosphorylation reaction shows strong dependence on MST2 concentration suggesting that it is an intermolecular reaction. While both the full-length MST2 and the caspase-3 truncated form of MST2 undergo autophosphorylation, the two forms of the phosphorylated MST2 display marked difference in susceptibility to protein phosphatases. The full-length phospho-MST2 is rapidly dephosphorylated by protein phosphatase 1 or protein phosphatase 2A whereas the truncated MST2 is remarkably resistant to the dephosphorylation. Based on the present results, a novel molecular mechanism for MST2 regulation in apoptotic cells is postulated. In normal cells, because of the low concentration and the ready reversal of the autophosphorylation by protein phosphatases, MST2 is present mainly in the unphosphorylated and inactive state. During cell apoptosis, MST2 is cleaved by caspase-3 and undergoes irreversible autophosphorylation, thus resulting in the accumulation of active MST2. PMID- 12554737 TI - A highly selective CCR2 chemokine agonist encoded by human herpesvirus 6. AB - The chemokine-like, secreted protein product of the U83 gene from human herpesvirus 6, here named vCCL4, was chemically synthesized to be characterized in a complete library of the 18 known human chemokine receptors expressed individually in stably transfected cell lines. vCCL4 was found to cause calcium mobilization as efficiently as the endogenous chemokine ligand CCL2 through the CCR2 receptor, whereas the virally encoded chemokine did not affect any of the other 17 human chemokine receptors tested. Mutual cross-desensitization between CCL2 and vCCL4 was demonstrated in the CCR2-transfected cells. The affinity of vCCL4 for the CCR2 receptor was 79 nm as determined in competition binding against radioactively labeled CCL2. In the murine pre-B lymphocyte cell line L1.2 stably transfected with the CCR2 receptor, vCCL4 acted as a relatively low potency but highly efficacious chemoattractant being equally or more efficacious in causing cell migration than CCL2 and CCL7 and considerably more efficacious than CCL8 and CCL13. It is concluded that human herpesvirus 6 encodes a highly selective and efficacious CCR2 agonist, which will attract CCR2 expressing cells, for example macrophages and monocytes, conceivably for the virus to infect and to establish latency in. It is suggested that vCCL4 during reactivation of the virus in for example monocyte-derived microglia could perhaps be involved in the pathogenesis of the CCR2-dependent disease, multiple sclerosis. PMID- 12554738 TI - Analysis of human flap endonuclease 1 mutants reveals a mechanism to prevent triplet repeat expansion. AB - Flap endonuclease 1 (FEN1), involved in the joining of Okazaki fragments, has been proposed to restrain DNA repeat sequence expansion, a process associated with aging and disease. Here we analyze properties of human FEN1 having mutations at two conserved glycines (G66S and G242D) causing defects in nuclease activity. Introduction of these mutants into yeast led to sequence expansions. Reconstituting triplet repeat expansion in vitro, we previously found that DNA ligase I promotes expansion, but FEN1 prevents the ligation that forms expanded products. Here we show that among the intermediates that could generate sequence expansion, a bubble is necessary for ligation to produce the expansion product. Severe exonuclease defects in the mutant FEN1 suggested that the inability to degrade bubbles exonucleolytically leads to expansion. However, even wild type FEN1 exonuclease cannot compete with DNA ligase I to degrade a bubble structure before it can be ligated. Instead, we propose that FEN1 suppresses sequence expansion by degrading flaps that equilibrate with bubbles, thereby reducing bubble concentration. In this way FEN1 employs endonuclease rather than exonuclease to prevent expansions. A model is presented describing the roles of DNA structure, DNA ligase I, and FEN1 in sequence expansion. PMID- 12554739 TI - Trapping HIV-1 reverse transcriptase before and after translocation on DNA. AB - A disulfide cross-linking strategy was used to covalently trap as a stable complex (complex N) a short-lived, kinetic intermediate in DNA polymerization. This intermediate corresponds to the product of polymerization prior to translocation. We also prepared the trapped complex that corresponds to the product of polymerization after translocation (complex P). The cross-linking method that we used is a variation of a technique developed by the Verdine and Harrison laboratories. It involves disulfide interchange between an engineered sulfhydryl group of the protein (Q258C mutation) and a disulfide-containing tether attached at the N(2) amino group of a modified dG in either the template or the primer strand of the nucleic acid. We report here a highly efficient synthesis of the precursor, bis(3-aminopropyl)disulfide dihydrochloride, used to introduce this substituent into the oligonucleotide. Efficient cross-linking takes place when the base pair containing the substituent is positioned seven registers from the dNTP-binding site (N site) and the N site is occupied. Complex N, but not complex P, is a substrate for the ATP-based excision reaction that unblocks nucleoside reverse transcriptase inhibitor (NRTI)-terminated primers and causes resistance to several NRTIs, confirming predictions that the excision reaction takes place only when the 3'-end of the primer is bound at the N site. These techniques can be used for biochemical and structural studies of the mechanism of DNA polymerization, translocation, and excision-based resistance of RT to NRTIs. They may also be useful in studying other DNA or RNA polymerases or other enzymes. PMID- 12554740 TI - Mapping of functional domains of gamma-SNAP. AB - gamma-Soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein (gamma SNAP) is capable of stabilizing a 20 S complex consisting of NSF, alpha-SNAP, and SNAP receptors (SNAREs), but its function in vesicular transport is not fully understood. Our two-hybrid analysis revealed that gamma-SNAP, unlike alpha-SNAP, interacts directly with NSF, as well as Gaf-1/Rip11, but not with SNAREs. Gaf 1/Rip11 is a gamma-SNAP-associated factor that belongs to the Rab11-interacting protein family. To gain insight into the molecular basis for the interactions of gamma-SNAP with NSF and Gaf-1/Rip11, we determined the regions of the three proteins involved in protein-protein interactions. gamma-SNAP bound to NSF via its extreme C-terminal region, and the full-length NSF was needed to interact with gamma-SNAP. Both the N-terminal and C-terminal regions of gamma-SNAP were required for the binding to Gaf-1/Rip11. Gaf-1/Rip11 bound to gamma-SNAP via its C-terminal domain comprising a putative coiled-coil region. Although the C terminal domain of Gaf-1/Rip11 also interacts with Rab11, the binding of gamma SNAP and Rab11 to Gaf-1/Rip11 was not mutually exclusive. Rather, Gaf-1/Rip11 was capable of serving a link between gamma-SNAP and Rab11. A complex comprising gamma-SNAP and Gaf-1/Rip11 was disassembled in a process coupled to NSF-mediated ATP hydrolysis, suggesting that the interaction between gamma-SNAP and Gaf 1/Rip11 is of functional significance. PMID- 12554741 TI - Deamidations in recombinant human phenylalanine hydroxylase. Identification of labile asparagine residues and functional characterization of Asn --> Asp mutant forms. AB - Recombinant human phenylalanine hydroxylase (hPAH) expressed in Escherichia coli for 24 h at 28 degrees C has been found by two-dimensional electrophoresis to exist as a mixture of four to five molecular forms as a result of nonenzymatic deamidation of labile Asn residues. The multiple deamidations alter the functional properties of the enzyme including its affinity for l-phenylalanine and tetrahydrobiopterin, catalytic efficiency, and substrate inhibition and also result in enzyme forms more susceptible to limited tryptic proteolysis. Asn(32) in the regulatory domain deamidates very rapidly because of its nearest neighbor amino acid Gly(33) (Solstad, T., Carvalho, R. N., Andersen, O. A., Waidelich, D., and Flatmark, T. (2003) Eur. J. Biochem., in press). Matrix-assisted laser desorption/ionization time of flight-mass spectrometry of the tryptic peptides in the catalytic domain of a 24-h (28 degrees C) expressed enzyme has shown Asn(376) and Asn(133) to be labile residues. Site-directed mutagenesis of nine Asn residues revealed that the deamidations of Asn(32) and Asn(376) are the main determinants for the functional and regulatory differences observed between the 2 and 24-h-induced wild-type (wt) enzyme. The Asn(32) --> Asp, Asn(376) --> Asp, and the double mutant forms expressed for 2 h at 28 degrees C revealed qualitatively similar regulatory properties as the highly deamidated 24-h expressed wt-hPAH. Moreover, deamidation of Asn(32) in the wt-hPAH (24 h expression at 28 degrees C) and the Asn(32) --> Asp mutation both increase the initial rate of phosphorylation of Ser(16) by cAMP-dependent protein kinase (p < 0.005). By contrast, the substitution of Gly(33) with Ala or Val, both preventing the deamidation of Asn(32), resulted in enzyme forms that were phosphorylated at a similar rate as nondeamidated wt-hPAH, even on 24-h expression. The other Asn - > Asp substitutions (in the catalytic domain) revealed that Asn(207) and Asn(223) have an important stabilizing structural function. Finally, two recently reported phenylketonuria mutations at Asn residues in the catalytic domain were studied, i.e. Asn(167) --> Ile and Asn(207) --> Asp, and their phenotypes were characterized. PMID- 12554742 TI - Prothymosin alpha is processed to thymosin alpha 1 and thymosin alpha 11 by a lysosomal asparaginyl endopeptidase. AB - Thymosin alpha(1) (T alpha(1)) and thymosin T alpha(11) (T alpha(11)) are polypeptides with immunoregulatory properties first isolated from thymic extracts, corresponding to the first 28 and 35 amino acid residues, respectively, of prothymosin alpha (ProT alpha), a protein involved in chromatin remodeling. It has been widely supposed that these polypeptides are not natural products of the in vivo processing of ProT alpha, since neither was found in extracts in which proteolysis was prevented. Here we show that a lysosomal asparaginyl endopeptidase is able to process ProT alpha to generate T alpha(1) and T alpha(11). In view of its catalytic properties and structural and immunological analyses, this protease was identified as mammalian legumain. It selectively cleaves some of the asparaginyl-glycine residues in the ProT alpha sequence; specifically, Asn(28)-Gly(29) and Asn(35)-Gly(36) residues are cleaved with similar efficiency in vitro to generate T alpha(1) and T alpha(11), respectively. By contrast T alpha(1) is the main product detected in vivo, free in the cytosol, at concentrations similar to that of ProT alpha. The data here reported demonstrate that T alpha(1) is not an artifact but rather is naturally present in diverse mammalian tissues and raise the possibility that it has a functional role. PMID- 12554743 TI - COL11A2 collagen gene transcription is differentially regulated by EWS/ERG sarcoma fusion protein and wild-type ERG. AB - A specific t(21;22) chromosomal translocation creates the chimeric EWS/ERG gene in some cases of Ewing's sarcoma. In the resultant EWS/ERG fusion protein, the N terminal part of the ETS family protein ERG is replaced by the N terminus of the RNA-binding protein EWS. We found that both the EWS/ERG and COL11A2 genes are expressed in the Ewing's sarcoma cell line, CADO-ES1. To investigate a potential role for EWS/ERG in COL11A2 gene expression, we characterized the COL11A2 promoter and tested the ability of wild-type ERG and EWS/ERG sarcoma fusion protein to transactivate COL11A2 promoter using a luciferase assay. We found that expression of EWS/ERG, but not wild-type ERG, transactivated the COL11A2 promoter and that this transactivation required not only the N-terminal region of EWS but also an intact DNA-binding domain from ERG. Electrophoretic mobility shift assay using COL11A2 promoter sequence showed involvement of EWS/ERG in the formation of DNA-protein complexes, and chromatin immunoprecipitation assay revealed direct interaction between COL11A2 promoter and EWS/ERG fusion protein in vivo. EWS/ERG, but not wild-type ERG, bound to RNA polymerase II. Treatment of cells with the histone deacetylase inhibitor trichostatin A enabled ERG to transactivate the COL11A2 promoter, therefore abolishing the differential effects of EWS/ERG and ERG. Taken together, these findings indicate that the COL11A2 gene is regulated both by potential ERG association with a histone deacetylase complex and by direct EWS/ERG recruitment of RNA polymerase II. PMID- 12554744 TI - Glucagon-like peptides: regulators of cell proliferation, differentiation, and apoptosis. AB - Peptide hormones are secreted from endocrine cells and neurons and exert their actions through activation of G protein-coupled receptors to regulate a diverse number of physiological systems including control of energy homeostasis, gastrointestinal motility, neuroendocrine circuits, and hormone secretion. The glucagon-like peptides, GLP-1 and GLP-2 are prototype peptide hormones released from gut endocrine cells in response to nutrient ingestion that regulate not only energy absorption and disposal, but also cell proliferation and survival. GLP-1 expands islet mass by stimulating pancreatic beta-cell proliferation and induction of islet neogenesis. GLP-1 also promotes cell differentiation, from exocrine cells or immature islet progenitors, toward a more differentiated beta cell phenotype. GLP-2 stimulates cell proliferation in the gastrointestinal mucosa, leading to expansion of the normal mucosal epithelium, or attenuation of intestinal injury in experimental models of intestinal disease. Both GLP-1 and GLP-2 exert antiapoptotic actions in vivo, resulting in preservation of beta-cell mass and gut epithelium, respectively. Furthermore, GLP-1 and GLP-2 promote direct resistance to apoptosis in cells expressing GLP-1 or GLP-2 receptors. Moreover, an increasing number of structurally related peptide hormones and neuropeptides exert cytoprotective effects through G protein-coupled receptor activation in diverse cell types. Hence, peptide hormones, as exemplified by GLP 1 and GLP-2, may prove to be useful adjunctive tools for enhancement of cell differentiation, tissue regeneration, and cytoprotection for the treatment of human disease. PMID- 12554745 TI - On the role of liver X receptors in lipid accumulation in adipocytes. AB - The pivotal role of liver X receptors (LXRs) in the metabolic conversion of cholesterol to bile acids in mice is well established. More recently, the LXRalpha promoter has been shown to be under tight regulation by peroxisome proliferator-activated receptors (PPARs), implying a role for LXRalpha in mediating the interplay between cholesterol and fatty acid metabolism. We have studied the role of LXR in fat cells and demonstrate that LXR is regulated during adipogenesis and augments fat accumulation in mature adipocytes. LXRalpha expression in murine 3T3-L1 adipocytes as well as in human adipocytes was up regulated in response to PPARgamma agonists. Administration of a PPARgamma agonist to obese Zucker rats also led to increased LXRalpha mRNA expression in adipose tissue in vivo. LXR agonist treatment of differentiating adipocytes led to increased lipid accumulation. An increase of the expression of the LXR target genes, sterol regulatory binding protein-1 and fatty acid synthase, was observed both in vivo and in vitro after treatment with LXR agonists for 24 h. Finally, we demonstrate that fat depots in LXRalpha/beta-deficient mice are smaller than in age-matched wild-type littermates. These findings imply a role for LXR in controlling lipid storage capacity in mature adipocytes and point to an intriguing physiological interplay between LXR and PPARgamma in controlling pathways in lipid handling. PMID- 12554746 TI - Pressure overload selectively up-regulates Ca2+/calmodulin-dependent protein kinase II in vivo. AB - Signals transduced by the multifunctional calcium/calmodulin-dependent protein kinases (CaMKs), have been suggested to regulate the development of hypertrophy. We address the role of the three multifunctional CaMKs, CaMK I, II, and IV, in this process using transverse aortic constriction (TAC) to induce cardiac hypertrophy in mice. We find a 33% increase in total CaMK activity 7 d after TAC. However, there are no changes in the levels of CaMKI, which is expressed in the ventricles, or CaMKIV, which is not detectable in the ventricles. Moreover, mice null for the CaMKIV gene develop ventricular hypertrophy and induce the expression of selected hypertrophy marker mRNAs, indicating that CaMKIV is not required at any time during the development of hypertrophy. On the other hand, TAC does increase both mRNA and protein levels of specific isoforms of CaMKII derived from both gamma and delta genes. Included among these isoforms are those that localize to both cytoplasm and nucleus. Collectively, the increased levels of CaMKII isoforms result in a constitutive increase in the Ca(2+)/calmodulin independent activity of CaMKII in the ventricles. We conclude that CaMKII is the multifunctional CaMK most likely to mediate Ca(2+)- dependent protein phosphorylation events in response to TAC-induced cardiac hypertrophy. PMID- 12554747 TI - Dynamic patterns of growth hormone gene transcription in individual living pituitary cells. AB - Real-time imaging of the GH gene promoter linked to luciferase in living pituitary cells has revealed surprising heterogeneity and variety of dynamic patterns of gene expression. Cells treated with either forskolin or thyroid hormone generated a consistent and characteristic temporal response from cell populations, but detailed analysis of individual cells revealed different patterns. Approximately 25-26% of cells displayed no response, 25-33% of cells exhibited a sustained progressive rise in luciferase activity, and 41-50% showed a transient phasic, or oscillatory response, after given stimuli. In cells treated consecutively with the two stimuli, the population response to the second stimulus was augmented. Single-cell analysis revealed that this was partly due to an increased number of cells responding, but also that the prevalence of response patterns changed: cells that responded to an initial stimulus were more likely to respond subsequently in a progressive sustained manner. In conclusion, these studies have indicated that GH promoter activity in individual living pituitary cells is unstable and possibly stochastic, with dynamic variations from hour to hour. The prevalence of different temporal patterns of response to hormonal stimulation among a population of cells is altered by the endocrine history of those cells. PMID- 12554748 TI - Estrogen receptor (ER)-beta reduces ERalpha-regulated gene transcription, supporting a "ying yang" relationship between ERalpha and ERbeta in mice. AB - Estrogen is of importance for the regulation of adult bone metabolism. The aim of the present study was to determine the role of estrogen receptor-beta (ERbeta) in vivo on global estrogen-regulated transcriptional activity in bone. The effect of estrogen in bone of ovariectomized mice was determined using microarray analysis including 9400 genes. Most of the genes (95% = 240 genes) that were increased by estrogen in wild-type (WT) mice were also increased by estrogen in ERbeta inactivated mice. Interestingly, the average stimulatory effect of estrogen on the mRNA levels of these genes was 85% higher in ERbeta-inactivated than in WT mice, demonstrating that ERbeta reduces estrogen receptor-alpha (ERalpha) regulated gene transcription in bone. The average stimulatory effect of estrogen on estrogen-regulated bone genes in ERalpha-inactivated mice was intermediate between that seen in WT and ERalphabeta double-inactivated mice. Thus, ERbeta inhibits ERalpha-mediated gene transcription in the presence of ERalpha, whereas, in the absence of ERalpha, it can partially replace ERalpha. In conclusion, our in vivo data indicate that an important physiological role of ERbeta is to modulate ERalpha-mediated gene transcription supporting a "Ying Yang" relationship between ERalpha and ERbeta in mice. PMID- 12554750 TI - Pem homeobox gene promoter sequences that direct transcription in a Sertoli cell specific, stage-specific, and androgen-dependent manner in the testis in vivo. AB - Although many genes are expressed selectively in Sertoli cells, regulatory sequences sufficient to drive Sertoli cell-specific expression in the postnatal and adult testis in vivo have not been identified. In the present study, we identified promoter sequences from the Pem homeobox gene that direct Sertoli cell specific expression in an androgen-dependent and stage-specific manner. Immunohistochemical and RNA analysis demonstrated that 0.6-kb 5'-flanking sequence directed transgene expression specifically in the testis and the epididymis but not in any other tissues tested. In the adult testis, this promoter fragment targeted the transgene expression specifically to Sertoli cells during stages IV-VIII of the seminiferous epithelial cycle, thereby mimicking the expression pattern of the endogenous Pem gene. This promoter fragment also recapitulated Pem's normal postnatal expression pattern, as it directed transcript induction between d 6 and d 9 post partum. Deletion of 0.3 kb from the 5'-end of the transgene had no effect on androgen-dependent Sertoli-specific expression but altered stage-specific expression in adult testes and caused premature postnatal expression. Our results suggest that there are at least two regulatory regions in the Pem proximal promoter: one that directs androgen receptor-dependent expression specifically in Sertoli cells within the testis and another that confers stage-specific expression in neonates and adults by acting as a negative regulator. To our knowledge, this is the first identification of regulatory regions that direct faithful developmentally regulated gene expression in postnatal and adult Sertoli cells in vivo. PMID- 12554749 TI - A PIT-1 homeodomain mutant blocks the intranuclear recruitment of the CCAAT/enhancer binding protein alpha required for prolactin gene transcription. AB - The pituitary-specific homeodomain protein Pit-1 cooperates with other transcription factors, including CCAAT/enhancer binding protein alpha (C/EBPalpha), in the regulation of pituitary lactotrope gene transcription. Here, we correlate cooperative activation of prolactin (PRL) gene transcription by Pit 1 and C/EBPalpha with changes in the subnuclear localization of these factors in living pituitary cells. Transiently expressed C/EBPalpha induced PRL gene transcription in pituitary GHFT1-5 cells, whereas the coexpression of Pit-1 and C/EBPalpha in HeLa cells demonstrated their cooperativity at the PRL promoter. Individually expressed Pit-1 or C/EBPalpha, fused to color variants of fluorescent proteins, occupied different subnuclear compartments in living pituitary cells. When coexpressed, Pit-1 recruited C/EBPalpha from regions of transcriptionally quiescent centromeric heterochromatin to the nuclear regions occupied by Pit-1. The homeodomain region of Pit-1 was necessary for the recruitment of C/EBPalpha. A point mutation in the Pit-1 homeodomain associated with the syndrome of combined pituitary hormone deficiency in humans also failed to recruit C/EBPalpha. This Pit-1 mutant functioned as a dominant inhibitor of PRL gene transcription and, instead of recruiting C/EBPalpha, was itself recruited by C/EBPalpha to centromeric heterochromatin. Together our results suggest that the intranuclear positioning of these factors determines whether they activate or silence PRL promoter activity. PMID- 12554751 TI - The critical role of carboxy-terminal amino acids in ligand-dependent and independent transactivation of the constitutive androstane receptor. AB - The mouse constitutive androstane receptor (CAR) is a unique member of the nuclear receptor superfamily, for which an inverse agonist, the testosterone metabolite 5alpha-androstan-3alpha-ol (androstanol), and an agonist, the xenobiotic 1,4-bis[2-(3, 5-dichloropyridyloxy)] benzene, are known. In this study the role of the transactivation domain 2 (AF-2) of CAR was investigated, which is formed by the seven most carboxy-terminal amino acids of the receptor. The AF-2 domain was shown to be critical for the constitutive activity by mediating a ligand-independent interaction of CAR with coactivator (CoA) proteins. In addition this domain increased and decreased contact with CoAs in the presence of agonist and inverse agonist, respectively. In analogy to classical endocrine nuclear receptors, in CAR the charge clamp between K187 (in helix 3) and E355 (within the AF-2 domain) was expected to be critical for its interaction with CoAs. However, the hydrophobic amino acids L352, L353, and I356 on the surface of the AF-2 domain were found to be more important for this protein-protein interaction. Moreover, these amino acids and C357 were shown to be involved in the response of CAR to androstanol. Interestingly, the cysteine at position 357 appears to block classical endocrine responsiveness of CAR to agonists, since mutagenesis of this amino acid both reduced CoA interaction in the absence of ligand and drastically increased inducibility by 1,4-bis[2-(3, 5 dichloropyridyloxy)] benzene. We showed that this blockade is not due to an intramolecular disulfide bridge, but is probably caused by an interaction between C357 and Y336. PMID- 12554752 TI - Molecular basis for the subtype discrimination of the estrogen receptor-beta selective ligand, diarylpropionitrile. AB - Although the two subtypes of the human estrogen receptor (ER), ERalpha and ERbeta, share only 56% amino acid sequence identity in their ligand binding domain (LBD), the residues that surround the ligand are nearly identical; nevertheless, subtype-selective ligands are known. To understand the molecular basis by which diarylpropionitrile (DPN), an ERbeta-selective ligand, is able to discriminate between the two ERs, we examined its activity on ER mutants and chimeric constructs generated by DNA shuffling. The N-terminal region of the ERbeta LBD (through helix 6) appears to be fully responsible for the ERbeta selectivity of DPN. In fact, a single ERalpha point mutation (L384M) was largely sufficient to switch the DPN response of this ER to that of the ERbeta type, but residues in helix 3 are also important in achieving the full ERbeta selectivity of DPN. Using molecular modeling, we found an energetically favorable fit for the S-DPN enantiomer in ERbeta, in which the proximal phenol mimics the A ring of estradiol, and the nitrile engages in stabilizing interactions with residues in the ligand-binding pocket of ERbeta. Our findings highlight that a limited number of critical interactions of DPN with the ERbeta ligand-binding pocket underlie its ER subtype-selective character. PMID- 12554753 TI - Bile acids induce the expression of the human peroxisome proliferator-activated receptor alpha gene via activation of the farnesoid X receptor. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) is a nuclear receptor that controls lipid and glucose metabolism and exerts antiinflammatory activities. PPARalpha is also reported to influence bile acid formation and bile composition. Farnesoid X receptor (FXR) is a bile acid-activated nuclear receptor that mediates the effects of bile acids on gene expression and plays a major role in bile acid and possibly also in lipid metabolism. Thus, both PPARalpha and FXR appear to act on common metabolic pathways. To determine the existence of a molecular cross-talk between these two nuclear receptors, the regulation of PPARalpha expression by bile acids was investigated. Incubation of human hepatoma HepG2 cells with the natural FXR ligand chenodeoxycholic acid (CDCA) as well as with the nonsteroidal FXR agonist GW4064 resulted in a significant induction of PPARalpha mRNA levels. In addition, hPPARalpha gene expression was up-regulated by taurocholic acid in human primary hepatocytes. Cotransfection of FXR/retinoid X receptor in the presence of CDCA led to up to a 3-fold induction of human PPARalpha promoter activity in HepG2 cells. Mutation analysis identified a FXR response element in the human PPARalpha promoter (alpha-FXR response element (alphaFXRE)] that mediates bile acid regulation of this promoter. FXR bound the alphaFXRE site as demonstrated by gel shift analysis, and CDCA specifically increased the activity of a heterologous promoter driven by four copies of the alphaFXRE. In contrast, neither the murine PPARalpha promoter, in which the alphaFXRE is not conserved, nor a mouse alphaFXRE-driven heterologous reporter, were responsive to CDCA treatment. Moreover, PPARalpha expression was not regulated in taurocholic acid-fed mice. Finally, induction of hPPARalpha mRNA levels by CDCA resulted in an enhanced induction of the expression of the PPARalpha target gene carnitine palmitoyltransferase I by PPARalpha ligands. In concert, these results demonstrate that bile acids stimulate PPARalpha expression in a species-specific manner via a FXRE located within the human PPARalpha promoter. These results provide molecular evidence for a cross-talk between the FXR and PPARalpha pathways in humans. PMID- 12554754 TI - Determination of nuclear receptor corepressor interactions with the thyroid hormone receptor. AB - The thyroid hormone receptor (TR) recruits the nuclear corepressors, nuclear receptor corepressor (NCoR) and silencing mediator of retinoid and thyroid hormone receptors (SMRT), to target DNA elements in the absence of ligand. While the TR preferentially recruits NCoR, the mechanism remains unclear. The corepressors interact with the TR via interacting domains (IDs) present in their C terminus which contain a conserved motif termed a CoRNR box. Despite their similarity, the corepressor IDs allow for nuclear receptor specificity. Here we demonstrate that NCoR stabilizes the TR homodimer when bound to DNA by preventing its dissociation from thyroid hormone response elements. This suggests that NCoR acts to hold the repression complex in place on target elements. The TR homodimer recruits NCoR through two of its three IDs, one of which is not present in SMRT. This unique ID, N3, contains a CoRNR box but lacks the extended helical motif present in each of the other IDs. Instead, N3 contains an isoleucine just proximal to this motif. This isoleucine is also conserved in N2 but not in the corresponding S2 domain in SMRT. On thyroid hormone response elements and in mammalian cells this residue is critical in both N3 and N2 for high-affinity TR binding. In addition, this residue also controls specificity for the interactions of TR with NCoR. Together these data suggest that the specific recruitment of NCoR by the TR through a unique motif allows for stabilization of the repression complex on target elements. PMID- 12554755 TI - Receptor-interacting protein 140 binds c-Jun and inhibits estradiol-induced activator protein-1 activity by reversing glucocorticoid receptor-interacting protein 1 effect. AB - In the presence of estradiol, estrogen receptor-alpha (ERalpha) increases transcription triggered by activator protein-1 (AP-1). We have previously shown that induction is mediated by the direct interaction between c-Jun and ERalpha, which stabilizes a multiprotein complex containing the coactivator GRIP1 (glucocorticoid receptor interacting protein 1). The effect of receptor interacting protein 140 (RIP140) in this regulation was assessed in the present study. We report that overexpression of RIP140 inhibits estradiol-induced AP-1 dependent transcription in a dose-dependent manner. Inhibition is not affected by trichostatin A, suggesting that histone deacetylase recruitment is not implicated. RIP140, which binds Jun proteins in pull-down assays and in intact cells, as shown by coimmunoprecipitation analysis and a mammalian one-hybrid system, participates in a multiprotein complex containing c-Jun and ERalpha. Moreover, the negative effect of RIP140 on AP-1-mediated transcription is relieved by GRIP1 overexpression and, conversely, RIP140 inhibits the stimulatory effect of GRIP1. The two cofactors compete for binding to c-Jun and ERalpha both in vitro and in intact cells, and GRIP1 interaction with both ERalpha and c-Jun is required for an efficient competition. These overall results suggest that the ratio between RIP140 and GRIP1 could determine, as proposed for hormone response element-mediated responses, the efficacy of estradiol in stimulating transcription of genes under AP-1 control. PMID- 12554756 TI - Endocrine-related resources from the National Institutes of Health. PMID- 12554758 TI - Insulin-like growth factor I receptor signaling and nuclear translocation of insulin receptor substrates 1 and 2. AB - The insulin receptor substrate 1 (IRS-1) can translocate to the nuclei and nucleoli of several types of cells. Nuclear translocation can be induced by an activated insulin-like growth factor 1 receptor (IGF-IR), and by certain oncogenes, such as the Simian virus 40 T antigen and v-src. We have asked whether IRS-2 could also translocate to the nuclei. In addition, we have studied the effects of functional mutations in the IGF-IR on nuclear translocation of IRS proteins. IRS-2 translocates to the nuclei of mouse embryo fibroblasts expressing the IGF-IR, but, at variance with IRS-1, does not translocate in cells expressing the Simian virus 40 T antigen. Mutations in the tyrosine kinase domain of the IGF IR abrogate translocation of the IRS proteins. Other mutations in the IGF-IR, which do not interfere with its mitogenicity but inhibit its transforming capacity, result in a decrease in translocation, especially to the nucleoli. Nuclear IRS-1 and IRS-2 interact with the upstream binding factor, which is a key regulator of RNA polymerase I activity and, therefore, rRNA synthesis. In 32D cells, wild-type, but not mutant, IRS-1 causes a significant activation of the ribosomal DNA promoter. The interaction of nuclear IRS proteins with upstream binding factor 1 constitutes the first direct link of these proteins with the ribosomal DNA transcription machinery. PMID- 12554759 TI - Transcription activation by the ecdysone receptor (EcR/USP): identification of activation functions. AB - The ecdysone receptor is a heterodimer of the two nuclear receptors EcR and ultraspiracle (USP). We have identified the regions of Drosophila EcR and USP responsible for transcriptional activation of a semisynthetic Eip71CD promoter in Kc cells. The isoform-specific A/B domains of EcR-B1 and B2, but not those of EcR A or USP, exhibit strong activation activity [activation function 1 (AF1)], both in isolation and in the context of the intact receptor. AF1 activity in isoform B1 derives from dispersed elements; the B2-specific AF1 consists of a 17-residue amphipathic helix. AF2 function was studied using a two-hybrid assay in Kc cells, based on the observation that potent hormone-dependent activation by the EcR/USP ligand-binding domain heterodimer requires the participation of both partners. Mutagenesis reveals that AF2 function depends on EcR helix 12, but not on the cognate USP region. EcR helix 12 mutants (F645A and W650A) exhibit a dominant negative phenotype. Thus, in the setting tested, the ecdysone receptor can activate transcription using the AF1 regions of EcR-B1 or -B2 and the AF2 region of EcR. USP acts as an allosteric effector for EcR, but does not contribute any intrinsic function. PMID- 12554760 TI - Gene expression profiling reveals progesterone-mediated cell cycle and immunoregulatory roles of Hoxa-10 in the preimplantation uterus. AB - Human infertility and recurrent pregnancy loss caused by implantation defects are poorly understood. Hoxa-10-deficient female mice have severe infertility and recurrent pregnancy loss due to defective uterine implantation. Gene expression profiling experiments reveal that Hoxa-10 is an important regulator of two critical events in implantation: stromal cell proliferation and local immunosuppression. At the time of implantation, Hoxa-10 mediates the progesterone stimulated proliferation of uterine stromal cells. Hoxa-10 mutants express a stromal cell proliferation defect that is accompanied by quantitative or spatial alterations in the expression of two cyclin-dependent kinase inhibitor genes, p57 and p15. Hoxa-10 deficiency also leads to a severe local immunological disturbance, characterized by a polyclonal proliferation of T cells, that occurs in place of the normal progesterone-mediated immunosuppression in the periimplantation uterus. PMID- 12554761 TI - Regulation of corticotropin-releasing hormone type 2 receptors by multiple promoters and alternative splicing: identification of multiple splice variants. AB - We demonstrate that multiple promoters and alternate splicing regulate expression of the human CRH receptor type 2 (CRHR2) gene. We show that flanking regions to the first exons drive promoter activity in both endogenously and nonendogenously expressing cell lines. Putative promoter elements have been identified that are conserved between species, including the comparison of CRHR2gamma in nonhuman primates that was previously known only in humans, which may be responsible for subtype tissue specific regulation. We have identified novel transcripts produced by alternate splicing of the first exon of CRHR2beta (beta1a) with various combinations of the 5' exons including a novel exon (beta1c) spliced to the common exons. The 5' structure of the gene permits many other combinations of alternate splicing that may arise as part of a regulatory mechanism controlling functional receptor expression. The 5'-untranslated region of the first exons has been extended; and 3' acceptor sites identified within the 5' untranslated region of CRHR2gamma and CRHR2alpha are used during alternate splicing of CRHR2beta upstream exons. This has important implications because various reports on the expression of CRHR2gamma and CRHR2alpha have been unable to discriminate between the functional receptor and CRHR2beta alternate splice variants. Only the described sequences upstream of the 3' splice site are unique to CRHR2gamma and CRHR2alpha. PMID- 12554762 TI - Estradiol stabilizes estrogen receptor messenger ribonucleic acid in sheep endometrium via discrete sequence elements in its 3'-untranslated region. AB - The preovulatory surge of estrogen up-regulates estrogen receptor-alpha (ER) gene expression in the uterus during the estrous/menstrual cycles of female mammals. Previously, we demonstrated that the 5-fold increase in ER mRNA levels in endometrium of ovariectomized ewes treated with a physiological dose of estradiol (E2) is entirely due to an increase in ER mRNA stability. Our current work confirms that the E2 effect is specific to ER mRNA. The sequence of ER mRNA, cloned from sheep endometrium, shows a high degree of conservation with those of other species, even in the 5'- and the very long 3'-untranslated regions. In a cell-free assay, ER mRNA demonstrates greater stability with endometrial extracts from E2-treated ewes compared with those from untreated ovariectomized ewes. The E2-enhanced stability of ER mRNA was ablated by prior treatment of the extracts with proteinase K, 70 C heat, and oxidizing and alkylating reagents, indicating that a protein is responsible for stabilization of the message. The 3' untranslated region of ER mRNA contains discrete sequences required for E2 enhanced stability, four of which were identified by extensive deletion mutant analyses. Transfer of two of the four minimal E2-modulated stability sequences conferred E2-enhanced stability to a heterologous RNA. These minimal E2-modulated stability sequences contain a common 10-base, uridine-rich sequence that is predicted to reside in a loop structure. Throughout our studies, estrogen stabilization of ER mRNA in sheep endometrium resembled that of vitellogenin mRNA in frog liver, indicating conservation of this ancient mechanism for enhancing gene expression in response to estrogen. PMID- 12554763 TI - Estrogen enhances depolarization-induced glutamate release through activation of phosphatidylinositol 3-kinase and mitogen-activated protein kinase in cultured hippocampal neurons. AB - Changes in synaptic efficacy are considered necessary for learning and memory. Recently, it has been suggested that estrogen controls synaptic function in the central nervous system. However, it is unclear how estrogen regulates synaptic function in central nervous system neurons. We found that estrogen potentiated presynaptic function in cultured hippocampal neurons. Chronic treatment with estradiol (1 or 10 nm) for 24 h significantly increased a high potassium-induced glutamate release. The estrogen-potentiated glutamate release required the activation of both phosphatidylinositol 3-kinase and MAPK. The high potassium evoked release with or without estradiol pretreatment was blocked by tetanus neurotoxin, which is an inhibitor of exocytosis. In addition, the reduction in intensity of FM1-43 fluorescence, which labeled presynaptic vesicles, was enhanced by estradiol, suggesting that estradiol potentiated the exocytotic mechanism. Furthermore, protein levels of synaptophysin, syntaxin, and synaptotagmin (synaptic proteins, respectively) were up-regulated by estradiol. We confirmed that the up-regulation of synaptophysin was blocked by the MAPK pathway inhibitor, U0126. These results suggested that estrogen enhanced presynaptic function through the up-regulated exocytotic system. In this study, we propose that estrogen reinforced excitatory synaptic transmission via potentiated-glutamate release from presynaptic sites. PMID- 12554764 TI - Consequences of single-chain translation on the structures of two chorionic gonadotropin yoked analogs in alpha-beta and beta-alpha configurations. AB - Human chorionic gonadotropin (hCG) is a placental-derived heterodimeric glycoprotein hormone, which, through the binding and activation of the LH receptor, rescues the corpus luteum and maintains pregnancy. The three dimensional structure of hCG is known; however, the relevance of its fold to bioactivity is unclear. Although both subunits (alpha and beta) are required for activity, recent data with single-chain analogs have suggested a diminished role for the cystine knot and an intact heterodimeric interface in binding and receptor activation in vitro. Herein, we report the purification and structural characterization of two yoked (Y) hCG analogs, YhCG1 (beta-alpha) and YhCG3 (alpha-beta). The fusion proteins yielded higher IC50s and EC50s than those of hCG; the maximal hCG-mediated cAMP production, however, was the same. Circular dichroic spectroscopy revealed that the three proteins exhibit distinct far UV circular dichroic spectra, with YhCG1 containing somewhat more secondary structure than YhCG3 and hCG. Limited proteolysis with proteinase K indicated that heterodimeric hCG was much more resistant to cleavage than the single-chain analogs. YhCG1 was more susceptible to proteolysis than YhCG3, and the fragmentation patterns were different in the two proteins. Taken together, the data presented herein provide direct structural evidence for altered three dimensional conformations in the two single-chain hCG analogs. Thus, the cognate G protein-coupled receptor can recognize and functionally respond to multiple ligand conformations. PMID- 12554765 TI - Insulin-like growth factor-I inhibits progesterone receptor expression in breast cancer cells via the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin pathway: progesterone receptor as a potential indicator of growth factor activity in breast cancer. AB - Although interactions between estrogen and growth factor signaling pathways have been studied extensively, how growth factors and progesterone regulate each other is less clear. In this study, we found that IGF-I sharply lowers progesterone receptor (PR) mRNA and protein levels in breast cancer cells. Other growth factors, such as epidermal growth factor, also showed the same effect. The decrease of PR levels was associated with reduced PR activity. Unlike progestins, IGF-I does not utilize the proteasome for down-regulating PR. Instead, the IGF-I mediated decrease in PR levels is via an inhibition of PR gene transcription. In addition, the phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway was found to be specifically involved in this IGF-I effect. Our data also suggest that the IGF-I down-regulation of PR is not mediated via a reduction of estrogen receptor (ER) levels or activity. First, IGF I induced ligand-independent ER activity while reducing ER-dependent PR levels. Second, whereas PR and cyclin D1 are both ER up-regulated, IGF-I increased cyclin D1 levels while decreasing PR levels. Third, constitutively active PI3K or Akt induced ER activity but reduced PR levels and activity. Taken together, our data indicate that IGF-I inhibits PR expression in breast cancer cells via the PI3K/Akt/mTOR pathway. Because low or absent PR in primary breast cancer is associated with poor prognosis and response to hormone therapy, our results suggest that low PR status may serve as an indicator of activated growth factor signaling in breast tumor cells, and therefore of an aggressive tumor phenotype and resistance against hormonal therapy. PMID- 12554766 TI - The NEDD8 pathway is required for proteasome-mediated degradation of human estrogen receptor (ER)-alpha and essential for the antiproliferative activity of ICI 182,780 in ERalpha-positive breast cancer cells. AB - Steroid hormone receptors, including estrogen receptor-alpha (ERalpha), are ligand-activated transcription factors, and hormone binding leads to depletion of receptor levels via preteasome-mediated degradation. NEDD8 (neural precursor cell expressed developmentally down-regulated) is an ubiquitin-like protein essential for protein processing and cell cycle progression. We recently demonstrated that ubiquitin-activating enzyme (Uba)3, the catalytic subunit of the NEDD8-activating enzyme, inhibits ERalpha transcriptional activity. Here we report that Uba3 mediated inhibition of ERalpha transactivation function is due to increased receptor protein turnover. Coexpression of Uba3 with ERalpha increased receptor degradation by the 26S proteasome. Inhibition of NEDD8 activation and conjugation diminished polyubiquitination of ERalpha and blocked proteasome-mediated degradation of receptor protein. The antiestrogen ICI 182,780 is known to induce ER degradation. In human MCF7 breast cancer cells modified to contain a disrupted NEDD8 pathway, ICI 182,780 degradation of ERalpha was impaired, and the antiestrogen was ineffective at inhibiting cell proliferation. This study provides the first evidence linking nuclear receptor degradation with the NEDD8 pathway and the ubiquitin-proteasome system, suggesting that the two pathways can act together to modulate ERalpha turnover and cellular responses to estrogens. Based on our observation that an intact NEDD8 pathway is essential for the antiproliferation activity of the ICI 182,780 in ERalpha positive breast cancer cells, we propose that disruptions in the NEDD8 pathway provide a mechanism by which breast cancer cells acquire antiestrogen resistance while retaining expression of ERalpha. PMID- 12554767 TI - Estradiol rapidly activates Akt via the ErbB2 signaling pathway. AB - Previously, we have demonstrated that the two mitogenic growth factors epidermal growth factor and IGF-I can activate Akt and estrogen receptor-alpha (ERalpha) in the hormone-dependent breast cancer cell line, MCF-7. In this report we now show that estradiol can also rapidly activate phosphatidylinositol 3-kinase (PI 3 K)/Akt and that this effect is mediated by the ErbB2 signaling pathway. Treatment of cells with estradiol resulted in phosphorylation of Akt and a 9-fold increase in Akt activity in 10 min. Akt activation was blocked by wortmannin and LY 294,002, two inhibitors of PI 3-K; by genistein, a protein tyrosine kinase inhibitor and an ER agonist; by AG825, a selective ErbB2 inhibitor; and by the antiestrogens ICI 182,780 and 4-hydroxy-tamoxifen; but not by rapamycin, an inhibitor of the ribosomal protein kinase p70S6K; nor by AG30, a selective epidermal growth factor receptor inhibitor. Akt activation by estradiol was abrogated by an arginine-to-cysteine mutation in the pleckstrin homology domain of Akt (R25C). Growth factors also activated Akt in the ER-negative variant of MCF-7, MCF-7/ADR, but estradiol did not induce Akt activity in these cells. Transient transfection of ERalpha into these cells restored Akt activation by estradiol, suggesting that estradiol activation of Akt requires the ERalpha. Estradiol did not activate Akt in MCF-7 cells stably transfected with an anti ErbB2-targeted ribozyme, further confirming a role for ErbB2. In vitro kinase assays using immunoprecipitation and anti-Akt1, -Akt2, and -Akt3-specific antibodies demonstrated that Akt1 is activated by estradiol in MCF-7 cells whereas Akt3 is the activated isoform in ER-negative MDA-MB231 cells, implying that selective activation of Akt subtypes plays a role in the actions of estradiol. Taken together, our data suggest that estradiol, bound to membrane ERalpha, interacts with and activates an ErbB dimer containing ErbB2, inducing activation of PI 3-K/Akt. PMID- 12554768 TI - A homogeneous in vitro functional assay for estrogen receptors: coactivator recruitment. AB - Estrogen receptor (ER)-mediated gene transcription occurs via the formation of a multimeric complex including ligand-activated receptors and nuclear coactivators. We have developed a homogeneous in vitro functional assay to help study the ligand-dependent interaction of ERs with various nuclear coactivators. The assay consists of FLAG-tagged ERalpha or ERbeta ligand binding domain (LBD), a biotinylated coactivator peptide, europium-labeled anti-FLAG antibody, and streptavidin-conjugated allophycocyanin. Upon agonist binding, the biotinylated coactivator peptide is recruited to FLAG-tagged ER LBD to form a complex and thus allow fluorescence resonance energy transfer (FRET) to occur between europium and allophycocyanin. Compounds with estrogen antagonism block the agonist-mediated recruitment of a coactivator and prevent FRET. The assay was used to evaluate the preference of ERs for various coactivators and ligands. Both ERalpha and ERbeta exhibited strong preferences for coactivator peptides corresponding to steroid receptor coactivator-1 and PPARgamma coactivor-1 vs. peroxisome proliferator activated receptor-interacting protein and cAMP response element binding protein binding protein. 17beta-Estradiol acted as a nonselective agonist for ERalpha and ERbeta. Genistein showed full agonism for ERalpha and only partial agonism for ERbeta, but with higher potency for ERbeta than ERalpha. Both raloxifene and tamoxifen behaved as full antagonists in this functional assay. The results obtained using compounds with a wide range of potency correlated well with those from a cell-based reporter gene assay. Therefore, this simple in vitro functional assay is predictive of ligand-dependent transactivation function of the receptor and, as such, is useful in nuclear receptor applications including mechanistic studies. PMID- 12554769 TI - Glucose-stimulated insulin secretion is coupled to the interaction of actin with the t-SNARE (target membrane soluble N-ethylmaleimide-sensitive factor attachment protein receptor protein) complex. AB - The actin monomer sequestering agent latrunculin B depolymerized beta-cell cortical actin, which resulted in increased glucose-stimulated insulin secretion in both cultured MIN6 beta-cells and isolated rat islet cells. In perifused islets, latrunculin B treatment increased both first- and second-phase glucose stimulated insulin secretion without any significant effect on total insulin content. This increase in secretion was independent of calcium regulation because latrunculin B also potentiated calcium-stimulated insulin secretion in permeabilized MIN6 cells. Confocal immunofluorescent microscopy revealed a redistribution of insulin granules to the cell periphery in response to glucose or latrunculin B, which correlated with a reduction in phalloidin staining of cortical actin. Moreover, the t-SNARE [target membrane soluble N-ethylmaleimide sensitive factor attachment protein (SNAP) receptor] proteins Syntaxin 1 and SNAP 25 coimmunoprecipitated polymerized actin from unstimulated MIN6 cells. Glucose stimulation transiently decreased the amount of actin coimmunoprecipitated with Syntaxin 1 and SNAP-25, and latrunculin B treatment fully ablated the coimmunoprecipitation. In contrast, the actin stabilizing agent jasplakinolide increased the amount of actin coimmunoprecipitated with the t-SNARE complex and prevented its dissociation upon glucose stimulation. These data suggest a mechanism whereby glucose modulates beta-cell cortical actin organization and disrupts the interaction of polymerized actin with the plasma membrane t-SNARE complex at a distal regulatory step in the exocytosis of insulin granules. PMID- 12554770 TI - Retinoic acid receptors beta and gamma do not repress, but instead activate target gene transcription in both the absence and presence of hormone ligand. AB - Retinoic acid receptors (RARs) are important mediators of retinoid signaling in morphogenesis, development, and cell differentiation. Three major isotypes of RARs, denoted alpha, beta, and gamma, have been identified, each encoded by a distinct genetic locus. Although RARalpha, RARbeta, and RARgamma share many structural and functional features, these three isotypes are known to play unique, as well as overlapping, roles in physiology and development. We report here that the three RAR isotypes display different transcriptional properties in the absence of hormone ligand; under these conditions, RARalpha is a strong repressor of target gene expression, whereas both RARbeta and RARgamma fail to repress and instead are able to mediate substantial levels of hormone-independent transcriptional activation. These differing transcriptional properties appear to reflect the differing abilities of the three RAR isotypes to interact with the SMRT (silencing mediator of retinoic acid and thyroid hormone receptor) corepressor protein: RARalpha binds to SMRT strongly both in vitro and in vivo, whereas RARbeta and RARgamma interact only weakly with SMRT. The ability to repress or to activate transcription in the absence of hormone maps predominantly to isotype-specific differences in the sequence of helix 3 within the hormone binding domain of the RARs, and the transcriptional properties of one isotype can be exchanged with that of another by exchanging portions of helix 3. The different transcriptional properties of RARalpha, RARbeta, and RARgamma in the absence of hormone contribute to the distinctive biological functions of these proteins and provide a rationale for the strong conservation of the three distinct isotypes during the vertebrate evolutionary radiation. PMID- 12554771 TI - Ras stimulates aberrant cell cycle progression and apoptosis in rat thyroid cells. AB - Abundant evidence supports the ability of Ras to stimulate thyroid cell proliferation. Stable expression of activated Ras enhances the sensitivity of thyroid cells to apoptosis. We report that apoptosis is a primary and general response of rat thyroid cells to acute expression of activated Ras in the absence or presence of thyrotropin, insulin, and serum, survival factors for thyroid cells. Ras induced apoptosis in quiescent and cycling cells. Concomitantly, Ras stimulated S phase entry in quiescent cells and enhanced G1/S transition in cycling cells. Ras effects on the cell cycle were characterized by delayed progression through S phase and an apparent failure to proceed through G2/M phase. Unlike thyroid cell mitogens, Ras markedly decreased cyclin D1 expression. Although acute expression of Ras decreased cyclin D1 protein levels, cells selected to survive chronic Ras expression exhibited a selective increase in cyclin D1 expression. In summary, thyroid cells harbor an apoptotic program activated by Ras that outstrips the protective effects of thyrotropin, insulin, and serum. Apoptosis is accompanied by dysregulated cell cycle progression, suggesting that cell death may arise, at least in part, as a consequence of inappropriate proliferative cues. PMID- 12554772 TI - Isoform-selective interactions between estrogen receptors and steroid receptor coactivators promoted by estradiol and ErbB-2 signaling in living cells. AB - Estrogen receptor (ER)alpha and -beta interact with a variety of coactivator proteins, most notably members of the steroid receptor coactivator (SRC) family, and these interactions have been shown to be regulated by estrogenic ligands and growth factor signaling. Here, using fluorescence resonance energy transfer (FRET), the selectivity of different stimulants on ERalpha and -beta interactions with coactivator receptor interaction domains (RIDs) were examined in living cells. We first show that ERalpha and ERbeta homo- and heterodimers form in vivo independently of the presence of 17beta-estradiol (E2) or antiestrogens. We then demonstrate that E2 enhances interactions between ERalpha and the RIDs of SRC-1 and SRC-3, whereas the interaction between ERalpha with the SRC-2 RID is ligand independent. The transcriptionally inactive mutant ERalphaL539A showed no interaction with all three SRC RIDs. Similarly, treatment with the antagonists 4 hydroxytamoxifen and EM-652 abolished all interactions between ERalpha and the SRC RIDs. FRET data also demonstrate that, in contrast to ERalpha, ERbeta interacts with all three SRC RIDs in a ligand-independent manner. However, these interactions were further enhanced or stabilized by E2, whereas the antiestrogen EM-652 abolished all interactions. In the presence of both ERalpha and ERbeta, E2 treatment led to the recruitment of SRC RIDs to the nuclei. Finally, expression of the oncogenic activated ErbB-2/Neu protein specifically enhanced ERalpha but not ERbeta interactions with SRC RIDs to an extent similar to E2-stimulated interactions. In summary, using FRET, we demonstrated preferential interactions between ER isoforms and coactivators upon hormonal treatment and activation of a growth factor signal transduction pathway in living cells. PMID- 12554773 TI - Dax-1 (dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene 1) gene transcription is regulated by wnt4 in the female developing gonad. AB - Dax-1 [dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene 1 (NR0B1)] is an orphan nuclear receptor acting as a suppressor of Ad4 binding protein/steroidogenic factor 1 [Ad4BP/SF-1 (NR5A1)] and as an anti-Sry factor in the process of gonadal sex differentiation. The roles of these nuclear receptors in the differentiation of the gonads and the adrenal cortex have been established through studies of the mutant phenotype in both mice and humans. However, the mechanisms underlying transcriptional regulation of these genes remain largely unknown. Here, we examined the relationship between Dax-1 gene transcription and the Wnt4 pathway. Reporter gene analysis revealed that Dax-1 gene transcription was activated by beta-catenin, a key signal transducing protein in the Wnt pathway, acting in synergy with Ad4BP/SF-1. Interaction between beta-catenin and Ad4BP/SF-1 was observed using yeast two hybrid and in vitro pull-down assays. The region of Ad4BP/SF-1 essential for this interaction consists of an acidic amino acid cluster, which resides in the first helix of the ligand-binding domain. Mutation of the amino acid cluster impaired transcriptional activation of Dax-1 as well as interaction of Ad4BP/SF-1 with beta-catenin. These results were supported by in vivo observations using Wnt4 gene-disrupted mice, in which Dax-1 gene expression was decreased significantly in sexually differentiating female gonads. We thus conclude that Wnt4 signaling mediates the increased expression of Dax-1 as the ovary becomes sexually differentiated. PMID- 12554774 TI - Bidirectional signaling between the estrogen receptor and the epidermal growth factor receptor. AB - Interactions between the estrogen receptor (ER) and the epidermal growth factor receptor (EGFR) contribute to the biological effects of these binding protein families. EGFR stimulates DNA synthesis and gene transcription in the uterus, related in part to estrogen-independent activation of the nuclear ER. This results from signal transduction enacted by the plasma membrane tyrosine kinase growth factor receptor, leading to 1) phosphorylation and activation of the nuclear ER, and 2) phosphorylation of coregulator proteins. More recently, it has been shown that a pool of ERalpha resides in or associates with the plasma membrane as a cytoplasmic protein. These ERs utilize the membrane EGFR to rapidly signal through various kinase cascades that influence both transcriptional and nontranscriptional actions of estrogen in breast cancer cells. This is congruent with a general theme of receptor signaling, where membrane G protein-coupled receptors activate tyrosine kinase growth factor receptors (EGFR, IGF-I receptor) that subsequently signal to MAPKs and other pathways. Overall, the bidirectional cross-talk between EGFR and cellular pools of ER contributes to reproductive organ physiology and pathophysiology. PMID- 12554775 TI - Short-term plasticity of cyclic adenosine 3',5'-monophosphate signaling in anterior pituitary corticotrope cells: the role of adenylyl cyclase isotypes. AB - Anterior pituitary corticotropes show a wide repertory of responses to hypothalamic neuropeptides and adrenal corticosteroids. The hypothesis that plasticity of the cAMP signaling system underlies this adaptive versatility was investigated. In dispersed rat anterior pituitary cells, depletion of intracellular Ca2+ stores with thapsigargin combined with ryanodine or caffeine enhanced the corticotropin releasing-factor (CRF)-evoked cAMP response by 4-fold, whereas reduction of Ca2+ entry alone had no effect. CRF-induced cAMP was amplified 15-fold by arginine-vasopressin (AVP) or phorbol-dibutyrate ester. In the presence of inhibitors of cyclic nucleotide phosphodiesterases and phorbol dibutyrate ester, the depletion of Ca2+ stores had no further effect on CRF induced cAMP accumulation. Adenohypophysial expression of mRNAs for the Ca2+ inhibited adenylyl cyclases (ACs) VI and IX, and the protein kinase C-stimulated ACs II and VII was demonstrated. ACIX was detected in corticotropes by immunocytochemistry, whereas ACII and ACVI were not present. The data show negative feedback regulation of CRF-induced cAMP levels by Ca2+ derived from ryanodine receptor-operated intracellular stores. Stimulation of protein kinase C by AVP enhances Ca2+-independent cAMP synthesis, thus changing the characteristics of intracellular Ca2+ feedback. It is proposed that the modulation of intracellular Ca2+ feedback in corticotropes by AVP is an important element of physiological control. PMID- 12554776 TI - Mitogen-activated protein kinase regulates nuclear association of human progesterone receptors. AB - Breast cancers often have increased MAPK activity; this pathway may drive breast cancer cell growth by targeting steroid hormone receptors. MAPK phosphorylates human progesterone receptors (PRs) on Ser294, thus regulating several aspects of PR activity. To study the role of PR Ser294 phosphorylation on subcellular distribution, we stably expressed wild-type (wt) or S294A (Ser294 to Ala) PR-B in several cell types. PRs phosphorylated on Ser294 were nuclear. Activation of MAPK induced Ser294 phosphorylation and rapid nuclear translocation of wt, but not S294A, PR-B; both receptors concentrated in the nucleus after progestin treatment. The MAPK kinase inhibitor, U0126, blocked epidermal growth factor but not progestin-induced Ser294 phosphorylation and translocation of wt PR, indicating a novel mechanism for nuclear localization. After progestin treatment, wt PR-B underwent ligand-dependent down-regulation, while S294A PR-B persisted in nuclei. Prolonged treatment with U0126 or the nuclear export inhibitor, leptomycin B, promoted nuclear accumulation of wt PR-B and blocked ligand dependent PR down-regulation, suggesting that PR degradation occurs in the cytoplasm and requires MAPK-dependent nuclear export. Stabilization of PRs by leptomycin B also blocked PR transcriptional activity, indicating a link between nucleocytoplasmic shuttling, receptor stability, and function. These results support a regulatory role for MAPK in nuclear steroid hormone receptor subcellular localization and coupling to multiple PR functions. PMID- 12554777 TI - A rapid, nongenomic, signaling pathway regulates the actin reorganization induced by activation of membrane testosterone receptors. AB - The human prostate cancer cell line LNCaP bears functional membrane testosterone receptors, which modify the actin cytoskeleton and increase the secretion of prostate-specific antigen (PSA) within minutes. Membrane steroid receptors are, indeed, a newly identified element of steroid action that is different from the classical intracellular sites. In the present work, using a nonpermeable analog of testosterone (testosterone-BSA), we investigated the signaling pathway that is triggered by the membrane testosterone receptors' activation and leads to actin cytoskeleton reorganization. We report that exposure of cells to testosterone-BSA resulted in phosphorylation of focal adhesion kinase (FAK), the association of FAK with the phosphatidylinositol-3 (PI-3) kinase, and the subsequent activation of the latter as well as the activation of the small guanosine triphosphatases Cdc42/Rac1. Pretreatment of cells with the specific PI-3 kinase inhibitor wortmannin abolished both the activation of the small guanosine triphosphatases and the alterations of actin cytoskeleton, whereas it did not affect the phosphorylation of FAK. These findings indicate that PI-3 kinase is activated downstream of FAK and upstream of Cdc42/Rac1, which subsequently regulate the actin organization. Moreover, wortmannin diminished the secretion of PSA, implying that the signaling events described above are responsible for the testosterone-BSA-induced PSA secretion. Our results are discussed under the prism of a possible implication of these membrane receptors in prostate cancer chemotherapy. PMID- 12554778 TI - Human pituitary tumor-transforming gene (PTTG1) motif suppresses prolactin expression. AB - Pituitary tumor-transforming gene (PTTG) originally isolated from GH-secreting pituitary adenoma cells causes in vitro cell transformation, in vivo tumorigenesis, and induces basic fibroblast growth factor. These functions require an intact C-terminal proline-proline-serine-proline motif. PTTG1 is abundantly expressed in human pituitary tumors and plays a role in the early stages of experimental prolactinoma formation. We now determined direct effects of PTTG1 on hormonal phenotypes of functional pituitary tumor cells. Overexpression of PTTG1 C terminus (amino acids 147-202) containing intact proline-proline-serine-proline motifs in rat prolactin (PRL)- and GH-secreting GH3 cells markedly abrogates PRL mRNA expression by more than 90% (P < 0.001) and hormone levels (P < 0.001) and PRL promoter activity (P < 0.01) compared with control vector cells or to a PTTG1 C terminus mutant (P163A, S165Q, P166L, P170L, P172A, and P173L). Wild-type PTTG1 C-terminal transfectants formed smaller (P < 0.05) sc tumors in rats compared with control or mutated PTTG1 C-terminal transfectants. Estrogen (10 nm) treatment for 48 h partially restored PRL expression in stable wild-type PTTG1 C-terminal transfectants. These results indicate that targeting PTTG1-mediated signaling alters the hormonal phenotype in pituitary cells and disrupted PTTG1 action may be a potential subcellular therapeutic tool for repressing PRL hypersecretion. PMID- 12554779 TI - Egr-1 induction in rat granulosa cells by follicle-stimulating hormone and luteinizing hormone: combinatorial regulation by transcription factors cyclic adenosine 3',5'-monophosphate regulatory element binding protein, serum response factor, sp1, and early growth response factor-1. AB - Early growth response factor (Egr-1) is an inducible zinc finger transcription factor that binds specific GC-rich enhancer elements and impacts female reproduction. These studies document for the first time that FSH rapidly induces Egr-1 expression in granulosa cells of small growing follicles. This response is transient but is reinitiated in preovulatory follicles exposed to the LH analog, human chorionic gonadotropin. Immunohistochemical analysis also showed gonadotropin induced Egr-1 in theca cells. The Egr-1 gene regulatory region responsive to gonadotropin signaling was localized within -164 bp of the transcription initiation site. Binding of Sp1/Sp3 to a proximal GC-box at -64/-46 bp was enhanced by FSH in immature granulosa cells but reduced after human chorionic gonadotropin stimulation of preovulatory follicles despite constant protein expression. This dynamic regulation of Sp1 binding was dependent on gonadotropin-regulated mechanisms that modulate Sp1/3-DNA binding activity. Serum response factor was active in granulosa cells and bound a consensus CArG box/serum response element site, whereas two putative cAMP response elements within the -164-bp region bound cAMP regulatory element (CRE) binding protein (CREB) and a second cAMP-inducible protein immunologically related to CREB. Transient transfection analyses using Egr-1 promoter-luciferase constructs and site-specific mutations show that the serum response element, GC-box, and CRE-131 are involved in gonadotropin regulation of Egr-1 expression in granulosa cells. Specific kinase inhibitors of Erk or protein kinase A antagonized this induction while exogenously expressed Egr-1 enhanced reporter expression. These observations indicate that the Egr-1 gene is a target of both FSH and LH action that may mediate molecular programs of proliferation and/or differentiation during follicle growth, ovulation, and luteinization. PMID- 12554780 TI - Regulation of the rat follicle-stimulating hormone beta-subunit promoter by activin. AB - FSH is controlled by a variety of positive and negative stimuli, and the unique FSHbeta-subunit is a major target for this regulation. Activin is a key modulator of FSHbeta transcription and hormone secretion. The signal transduction pathway leading to FSH expression was previously unknown. Here, we show that the transcription factors Smad3 and Smad4 mediate activin-stimulated activity of the rat FSHbeta promoter in a pituitary-derived cell line, LbetaT2. Cells were transiently transfected with the rat FSHbeta promoter fused to a luciferase reporter gene (-338rFSHbeta-Luc), and a minimal activin-responsive region was identified. Transfection of Smad3, but not the highly related Smad2, led to a ligand-independent stimulation of the FSHbeta promoter activity. As expected, activin caused an additional increase of luciferase expression, which was blocked by cotreatment with follistatin. Although Smad4 alone had no effect on FSHbeta transcription, it significantly augmented Smad3 and activin-mediated stimulation of the promoter. A palindromic consensus Smad-binding element in the proximal promoter was found to bind Smad4, and elimination of the region resulted in a loss of activin-mediated FSHbeta transcription. The activin signaling pathway is conserved in a number of cells, but FSHbeta expression is restricted to gonadotropes. A pituitary-specific transcription factor necessary for activin dependent induction of the FSHbeta promoter has been identified that permits FSHbeta expression in nongonadotrope cells. Pitx2 is a member of Pitx subfamily of bicoid-related homeodomain factors that is required for pituitary development and is present in the adult pituitary. This factor was transfected into LbetaT2 cells, where it caused up-regulation of basal and activin-mediated FSHbeta promoter activity. Furthermore, cotransfection of Pitx2c with Smad3 in kidney derived TSA cells resulted in activin-regulated FSHbeta response, suggesting its important role in tissue-restricted regulation of FSHbeta by activin. A Pitx2c binding site was identified within the proximal promoter, and elimination of this region also resulted in a loss of activin-regulated FSHbeta promoter activity. Taken together, these studies suggest that the regulation of FSHbeta is dependent on activin-mediated signaling factors in concert with pituitary-derived nuclear regulatory proteins. PMID- 12554781 TI - Regulation of niemann-pick c1 gene expression by the 3'5'-cyclic adenosine monophosphate pathway in steroidogenic cells. AB - The Niemann Pick-C1 (NPC-1) protein is essential for intracellular transport of cholesterol derived from low-density lipoprotein import in mammalian cells. The role of the protein kinase A (PKA) pathway in regulation of expression of the NPC 1 gene was investigated. NPC-1 promoter activity was induced by treatment with dibutryl cAMP (dbcAMP), alone or in combination with the cAMP response element (CRE) binding protein (CREB) overexpressed in adrenal Y-1 cells. When the catalytic subunit of PKA was overexpressed in Y-1 cells, there were similar increases in NPC-1 promoter activity in the presence of CREB. Responses were attenuated by blockade of the PKA pathway, and in the Kin-8 cell line deficient in PKA. Promoter deletion analysis revealed that this response was present in promoter fragments of 186 bp and larger but not present in the 121-bp fragment. Two promoter regions, one at -430 and one at -120 upstream of the translation initiation site, contained CRE consensus sequences. These bound recombinant CREB in EMSA, confirming their authenticity as CREB response elements. Promoters bearing mutations of both CRE displayed no response to dbcAMP. The orphan nuclear receptor, steroidogenic factor-1 (SF-1), was implicated in NPC-1 transactivation by the presence of SF-1 target sequence that formed a complex with recombinant SF 1 in EMSA. Furthermore, transfection of a plasmid that overexpressed SF-1 into ovarian granulosa cells increased promoter activity in response to dbcAMP, an effect abrogated by mutation of the SF-1 target sequence. Chromatin immunoprecipitation assays demonstrated that the CRE region of the endogenous and transfected NPC-1 promoter associated with both acetylated and phosphorylated histone H-3 and that this association was increased by dbcAMP treatment. Treatment with dbcAMP also increased the association of the CRE region of the promoter with CREB binding protein, which has histone acetyltransferase activity. Together, these results demonstrate a mechanism of regulation of NPC-1 expression by the cAMP-PKA pathway that includes PKA phosphorylation of CREB, recruitment of the coactivator CREB binding protein and the phosphorylation and acetylation of histone H-3 to transactivate the NPC-1 promoter. PMID- 12554782 TI - Two resistance to thyroid hormone mutants with impaired hormone binding. AB - Resistance to hormones is commonly due to mutations in genes encoding receptors. Resistance to thyroid hormone is due mostly to mutations of the beta-form of the human (h) thyroid hormone receptor (hTRbeta). We determined x-ray crystal structures of two hTRbeta ligand-binding domains (LBDs), Ala 317 Thr and Arg 316 His. Amino acids 316 and 317 form part of the hormone-binding pocket. The methyl of Ala 317, contacting iodine, sculpts the T3 hormone-binding pocket. Arg 316 is not in direct contact with T3 and has an unknown role in function. Remarkably, the Arg forms part of an unusual buried polar cluster in hTRbeta. Although the identity of the amino acids changes, the polar cluster appears in all nuclear receptors. In spite of the differing roles of 316 and 317, both resistance to thyroid hormone mutants display decreased T3 affinity and weakened transcriptional activation. The two mutants differ in that the Arg 316 His receptor does not form TR-TR homodimers on DNA. 3,5,3'-Triiodothyroacetic acid is bound to both receptors. Thr 317 repositions 3,5,3'-triiodothyroacetic acid distending the face of the receptor that binds coregulators. Arg 316 forms two hydrogen bonds with helix 1. Both are lost with mutation to His displacing helix 1 of the LBD and disordering the loop after helix 1. The stability of the helix 1, deriving in part from the buried polar cluster, is important for hormone binding and formation of TR dimers. The observation that the Arg 316 His mutation affects these functions implies a role for helix 1 in linking hormone binding to the DNA-binding domain-LBD configuration. PMID- 12554783 TI - Transcriptional induction of the osteocalcin gene during osteoblast differentiation involves acetylation of histones h3 and h4. AB - The remodeling of chromatin is required for tissue-specific gene activation to permit interactions of transcription factors and coregulators with their cognate elements. Here, we investigate the chromatin-mediated mechanisms by which the bone-specific osteocalcin (OC) gene is transcriptionally activated during cessation of cell growth in ROS 17/2.8 osteosarcoma cells and during normal osteoblast differentiation. Acetylation of histones H3 and H4 at the OC gene promoter was assayed during the proliferative and postproliferative stages of cell growth by using chromatin immunoprecipitation assays with antibodies that recognize different acetylated forms of histones H3 or H4. The results show that the promoter and coding regions of the OC gene contain very low levels of acetylated histones H3 and H4 during the proliferative period of osteoblast differentiation when the OC gene is inactive. Active expression of the OC gene in mature osteoblasts and confluent ROS 17/2.8 cells is functionally linked to preferential acetylation of histone H4 and, to a lesser extent, to acetylation of histone H3. Histone acetylation at the loci for RUNX2 (CBFA1), alkaline phosphatase, bone sialoprotein, osteopontin, and the cell growth regulator p21, which are expressed throughout osteoblast differentiation, is not altered postproliferatively. We conclude that acetylation of histones H3 and H4 is functionally coupled to the chromatin remodeling events that mediate the developmental induction of OC gene transcription in bone cells. PMID- 12554784 TI - Three mitogen-activated protein kinases inhibit insulin signaling by different mechanisms in 3T3-L1 adipocytes. AB - TNFalpha, which activates three different MAPKs [ERK, p38, and jun amino terminal kinase (JNK)], also induces insulin resistance. To better understand the respective roles of these three MAPK pathways in insulin signaling and their contribution to insulin resistance, constitutively active MAPK/ERK kinase (MEK)1, MAPK kinase (MKK6), and MKK7 mutants were overexpressed in 3T3-L1 adipocytes using an adenovirus-mediated transfection procedure. The MEK1 mutant, which activates ERK, markedly down-regulated expression of the insulin receptor (IR) and its major substrates, IRS-1 and IRS-2, mRNA and protein, and in turn reduced tyrosine phosphorylation of IR as well as IRS-1 and IRS-2 and their associated phosphatidyl inositol 3-kinase (PI3K) activity. The MKK6 mutant, which activates p38, moderately inhibited IRS-1 and IRS-2 expressions and IRS-1-associated PI3K activity without exerting a significant effect on the IR. Finally, the MKK7 mutant, which activates JNK, reduced tyrosine phosphorylation of IRS-1 and IRS-2 and IRS-associated PI3K activity without affecting expression of the IR, IRS-1, or IRS-2. In the context of our earlier report showing down-regulation of glucose transporter 4 by MEK1-ERK and MKK6/3-p38, the present findings suggest that chronic activation of ERK, p38, or JNK can induce insulin resistance by affecting glucose transporter expression and insulin signaling, though via distinctly different mechanisms. The contribution of ERK is, however, the strongest. PMID- 12554785 TI - Imaging the localized protein interactions between Pit-1 and the CCAAT/enhancer binding protein alpha in the living pituitary cell nucleus. AB - The homeodomain protein Pit-1 cooperates with the basic-leucine zipper protein CCAAT/enhancer binding protein alpha (C/EBPalpha) to control pituitary-specific prolactin gene transcription. We previously observed that C/EBPalpha was concentrated in regions of centromeric heterochromatin in pituitary GHFT1-5 cells and that coexpressed Pit-1 redistributed C/EBPalpha to the subnuclear sites occupied by Pit-1. Here, we used fluorescence resonance energy transfer microscopy to show that when C/EBPalpha was recruited by Pit-1, the average distance separating the fluorophores labeling the proteins was less than 7 nm. A mutation in the Pit-1 homeodomain, or truncation of the C/EBPalpha transactivation domain disrupted the redistribution of C/EBPalpha by Pit-1. Fluorescence resonance energy transfer analysis revealed that the mutant Pit-1 still associated with C/EBPalpha, and the truncated C/EBPalpha still associated with Pit-1, but these interactions were preferentially localized in regions of centromeric heterochromatin. In contrast, a truncation in C/EBPalpha that prevented DNA binding also blocked its association with Pit-1, suggesting that the binding of C/EBPalpha to DNA is a critical first step in specifying its association with Pit-1. These findings indicated that the protein domains that specify the interaction of Pit-1 and C/EBPalpha are separable from the protein domains that direct the positioning of the associated proteins within the nucleus. The intimate association of Pit-1 and C/EBPalpha at certain sites within the living cell nucleus could foster their combinatorial activities in the regulation of pituitary-specific gene expression. PMID- 12554786 TI - Dynamic inhibition of nuclear receptor activation by corepressor binding. AB - Nuclear receptors adopt dramatically different conformations in the presence or absence of ligand, and such liganded (holo) and unliganded (apo) receptors are specifically recognized by transcriptional coactivators and corepressors, respectively. These two states likely exist in dynamic equilibrium, contrary to the conventional model of static off and on conformations. First, corepressor SMRT [for silencing mediator of thyroid hormone receptor (TR) and retinoic acid receptor (RAR)] inhibits the interaction of coactivator steroid receptor coactivator-1 with liganded TR/RAR. Second, SMRT enables receptors to adopt apo form even in the presence of ligand, as demonstrated with limited proteolyses and decreased binding of radiolabeled retinoid to RAR. Finally, chromatin immunoprecipitation results indicate that SMRT and steroid receptor coactivator-1 dynamically compete for receptor bindings in vivo in the presence of ligand. These results suggest that corepressor binding can drive receptors to adopt the apo-state, even in the presence of ligand, and inhibit activated liganded (holo) nuclear receptors in vivo. PMID- 12554787 TI - Identification of a transferable two-amino-acid motif (GT) present in the C terminal tail of the human lutropin receptor that redirects internalized G protein-coupled receptors from a degradation to a recycling pathway. AB - Although highly homologous in amino acid sequence, the agonist-receptor complexes formed by the human lutropin receptor (hLHR) and rat (r) LHR follow different intracellular routes. The agonist-rLHR complex is routed mostly to a lysosomal degradation pathway whereas a substantial portion of the agonist-hLHR complex is routed to a recycling pathway. In a previous study, we showed that grafting a five-residue sequence (GTALL) present in the C-terminal tail of the hLHR into the equivalent position of the rLHR redirects a substantial portion of the internalized agonist-rLHR complex to a recycling pathway. Using a number of mutations of the GTALL motif, we now show that only the first two residues (GT) of this motif are necessary and sufficient to induce recycling of the internalized agonist-rLHR complex. Phosphoamino acid analysis and mutations of the GT motif show that phosphorylation of the threonine residue is not necessary for recycling. Lastly, we show that addition of portions of the C-terminal tail of the hLHR that include the GT motif to the C-terminal tails of the rat follitropin or murine delta-opioid receptors promotes the post-endocytotic recycling of these G protein-coupled receptors.We conclude that the GT motif present in the C-terminal tail of the hLHR is a transferable motif that promotes the postendocytotic recycling of several G protein-coupled receptors and that the GT-induced recycling does not require the phosphorylation of the threonine residue. PMID- 12554788 TI - A role for thyroid hormone transporters in transcriptional regulation by thyroid hormone receptors. AB - Thyroid hormones (THs) must be taken up by target cells to act at the genomic level through binding to nuclear thyroid hormone receptors (TRs). Extensive study has been made of mechanisms by which TH-bound TRs regulate transcription, yet little is known about the critical upstream step, i.e. how THs enter the cell. Growing evidence suggests that saturable transport mechanisms mediate the greater part of TH movement across the plasma membrane and have important roles in the regulation of TH bioavailability. For example, System L is a multifunctional transport system serving as a plasma membrane transporter of THs and amino acids in mammalian cells. We have used two complementary systems, the Xenopus oocyte (which has negligible basal System L activity) and the mammalian BeWo cell line (which has System L activity for TH transport), to investigate the role of this representative TH transporter in nuclear action of THs. We demonstrate that overexpression of System L in Xenopus oocytes increases both cytoplasmic and nuclear delivery of THs from external medium and also enhances transcriptional activation by TRs. Conversely, blocking endogenous System L activity in BeWo cells with specific inhibitors reduces both TH uptake and TR function. These results indicate that plasma membrane TH transporters such as System L may have important roles in gene regulation by TRs. PMID- 12554789 TI - Follicle-stimulating hormone increases testicular Anti-Mullerian hormone (AMH) production through sertoli cell proliferation and a nonclassical cyclic adenosine 5'-monophosphate-mediated activation of the AMH Gene. AB - Anti-Mullerian hormone (AMH) production by testicular Sertoli cells is high before puberty and can be further induced by FSH. Our objective was to delineate the mechanisms by which FSH stimulates AMH production. Assay of serum AMH levels and histological morphometric analysis in prepubertal FSH-deficient transgenic mice showed that serum AMH and testicular mass were decreased owing to reduced Sertoli cell number. All parameters resumed normal values in mice treated with recombinant FSH. We also analyzed the ability of FSH and the factors involved in its signaling pathway to activate AMH transcription by transfecting AMH promoter luc reporter constructs of different lengths in a prepubertal Sertoli cell line. Our results showed that FSH activates AMH transcription via adenylate cyclase, cAMP, and protein kinase A but involving a nonclassical cAMP-response pathway requiring nuclear factor-kappaB and activating protein 2 binding sites, which lie more than 1.9 kb upstream of the AMH transcription start site. This is the first report showing the importance of distant sequences in the regulation of AMH expression. We conclude that prepubertal testicular AMH production is increased by FSH stimulation through Sertoli cell proliferation and an enhancement of AMH gene transcription. PMID- 12554790 TI - Protein tyrosine phosphatase PTP20 induces actin cytoskeleton reorganization by dephosphorylating p190 RhoGAP in rat ovarian granulosa cells stimulated with follicle-stimulating hormone. AB - We identified 25 protein tyrosine phosphatases (PTPs) expressed in rat ovarian granulosa cells. Of these PTPs, the expression levels of at least PTP20, PTP MEG1, PTPepsilonM, and PTPepsilonC significantly changed during the estrous cycle. We examined the cellular functions of PTP20 in granulosa cells by expressing the wild type, a catalytically inactive CS mutant in which Cys229 of PTP20 was changed to Ser, or a substrate-trapping DA mutant in which Asp197 was mutated to Ala, using an adenovirus vector. Overexpression of the wild type, but not of the CS mutant, induced retraction of the cell body with the extension of long, dendritic-like processes after stimulation with FSH, a critical factor for the survival and differentiation of these cells. In addition, cell adhesion to the substratum decreased in an FSH-dependent manner. Inhibiting Rho GTPase activity with C3 botulinum toxin caused similar morphological changes. The FSH enhanced phosphotyrosine (p-Tyr) level of p190 RhoGAP was selectively reduced by the overexpressed wild type, but not by mutated PTP20. Although p190 RhoGAP is tyrosine phosphorylated by c-Src via the tyrosine kinase Pyk2, wild-type PTP20 had little effect on p-Tyr418 of c-Src and no effect on p-Tyr402 of Pyk2, which are required for full c-Src activity and for interacting between Pyk2 and c-Src, respectively. The CS and DA mutants as well as the wild type reduced the formation of p190 RhoGAP-p120 RasGAP complexes. Confocal microscopy analysis revealed that PTP20 intracellularly colocalizes with p190 RhoGAP. These results demonstrate that PTP20 regulates the functions of granulosa cells in an FSH dependent manner by dephosphorylating p190 RhoGAP and subsequently inducing reorganization of the actin cytoskeleton. Moreover, our data suggest that PTPs play significant roles in controlling the dynamics of ovarian functions. PMID- 12554791 TI - Local insulin-like growth factor-II mediates prolactin-induced mammary gland development. AB - Prolactin (PRL) is a major determinant of mammary epithelial cell proliferation during alveolar development in sexually mature and pregnant mice. To date, it has not been clear whether PRL effects these responses alone or by also invoking the action of autocrine/paracrine growth factors. In this study, we provide evidence that part of the effect of PRL on mammary gland growth is mediated by IGF-II. During sexual maturity and in early pregnancy, the level of IGF-II mRNA in the mammary gland was increased concurrent with increased PRL receptor expression. The level of IGF-II mRNA was reduced in mammary tissue from PRL receptor-/- mice during early pregnancy, and explants of mouse mammary gland and HC11 mammary epithelial cells both increased their expression of IGF-II after exposure to PRL in vitro. These findings coincided with the demonstration that IGF-II stimulated alveolar development in mammary glands in whole organ culture. PRL was most efficacious in stimulating IGF-II gene transcription from promoter 3 of the mouse IGF-II gene in vitro. Insight into the mechanism by which PRL induced IGF-II expression was provided by the fact that it was blocked by the Jak2 inhibitor AG490 and the MAPK inhibitor PD98059. Finally, induction of insulin receptor substrate (IRS)-1 in the mammary glands of PRL-treated mice and induction of IRS 1 and IRS-2 after treatment with PRL plus progesterone indicates that these molecules are induced by PRL as potential signaling intermediates downstream from IGF-I/insulin receptors. Together, these data demonstrate a role for IGF-II as a mediator of PRL action in the mouse mammary gland during ductal branching and alveolar development. PMID- 12554792 TI - Distinct properties and advantages of a novel peroxisome proliferator-activated protein [gamma] selective modulator. AB - Antidiabetic thiazolidinediones (TZDs) and non-TZD compounds have been shown to serve as agonists of the peroxisome proliferator-activated receptor gamma (PPARgamma). Here, we report the identification and characterization of a novel non-TZD selective PPARgamma modulator (nTZDpa). nTZDpa bound potently to PPARgamma with high selectivity vs. PPARalpha or PPARdelta. In cell-based assays for transcriptional activation, nTZDpa served as a selective, potent PPARgamma partial agonist and was able to antagonize the activity of PPARgamma full agonists. nTZDpa also displayed partial agonist effects when its ability to promote adipogenesis in 3T3-L1 cells was evaluated. Assessment of protein conformation using protease protection or solution nuclear magnetic resonance spectroscopy methods showed that nTZDpa produced altered PPARgamma conformational stability vs. full agonists, thereby establishing a physical basis for its observed partial agonism. DNA microarray analysis of RNA from 3T3-L1 adipocytes treated with nTZDpa or several structurally diverse PPARgamma full agonists demonstrated qualitative differences in the affected gene expression profile for nTZDpa. Chronic treatment of fat-fed, C57BL/6J mice with nTZDpa or a TZD full agonist ameliorated hyperglycemia and hyperinsulinemia. However, unlike the TZD, nTZDpa caused reductions in weight gain and adipose depot size. Feed efficiency was also substantially diminished. Unlike TZDs, nTZDpa did not cause cardiac hypertrophy in mice. When a panel of PPARgamma target genes was examined in white adipose tissue, nTZDpa produced a different in vivo expression pattern vs. the full agonist. These findings establish that novel selective PPARgamma modulators can produce altered receptor conformational stability leading to distinctive gene expression profiles, reduced adipogenic cellular effects, and potentially improved in vivo biological responses. Such compounds may lead to preferred therapies for diabetes, obesity, or metabolic syndrome. PMID- 12554793 TI - Oxytocin and vasopressin V1a and V2 receptors form constitutive homo- and heterodimers during biosynthesis. AB - G protein-coupled receptor (GPCR) oligomerization is a growing concept that has emerged from several studies suggesting that GPCRs can form both homo- and heterodimers. Using both coimmunoprecipitation and bioluminescence resonance energy transfer (BRET) approaches, we established that the vasopressin V1a, V2, and the oxytocin receptors exist as homo- and hetero-dimers in transfected human embryonic kidney 293T cells. Each receptor protomer had a similar propensity to form homo- and heterodimers, indicating that their relative expression levels may determine the homo-/heterodimer ratio. The finding that immature forms of the receptor can be immunoprecipitated as homo- and heterodimers and the detection by BRET of such oligomer in endoplasmic reticulum-enriched fractions suggest that the oligomerization processes take place early during biosynthesis. Treatment with agonists or antagonists did not modify the BRET among any of the vasopressin and oxytocin receptor pairs studied, indicating that the dimerization state of the receptors is not regulated by ligand binding once they have reached the cell surface. Taken together, these results strongly support the notion that GPCR dimerization is a constitutive process. PMID- 12554794 TI - Parathyroid hormone bone anabolic action requires Cbfa1/Runx2-dependent signaling. AB - The Cbfa1/Runx2 (referred to herein as Cbfa1) transcription factor has been shown to be essential for osteoblast differentiation and bone formation during embryogenesis. PTH given intermittently is a proven bone anabolic agent. Here, we investigated whether PTH regulates the expression and/or activity of Cbfa1 in osteoblastic cells and in a rat metatarsal organ culture assay. PTH was found to regulate Cbfa1 mRNA in the rat osteosarcoma cell line UMR106 in a concentration dependent manner. The effect of PTH was mimicked by forskolin, an activator of adenylate cyclase leading to the protein kinase A pathway. PTH administered intermittently for 5 d in vivo was found to stimulate Cbfa1 protein in the rat proximal tibiae metaphysis. To demonstrate PTH regulation of Cbfa1 activity, a construct containing six tandem Cbfa1 binding elements fused to luciferase was shown to be rapidly stimulated in response to PTH. This stimulation preceded the effects on mRNA regulation and resulted from a protein kinase A-mediated increase in Cbfa1 activity. Finally, using a neonate rat metatarsal organ culture system, we demonstrated dose-dependent anabolic responsiveness to PTH and to Cbfa1 overexpression from an adenoviral construct. We further showed that Cbfa1 antisense oligonucleotides that blocked adenoviral Cbfa1-induced anabolic effects in this organ culture model also abolished the PTH-mediated anabolic increase. These findings suggest a requirement for Cbfa1 in mediating the anabolic effects of PTH. Thus, regulation of Cbfa1 expression or activity is an important mechanism by which PTH controls osteoblast function. PMID- 12554795 TI - Differential regulation of rat and human CYP7A1 by the nuclear oxysterol receptor liver X receptor-alpha. AB - In rodent liver, transcription of the gene encoding cholesterol 7alpha hydroxylase (CYP7A1), which catalyzes the rate-limiting step in the classic bile acid synthetic pathway, is stimulated by the liver X receptor alpha (LXRalpha), a nuclear receptor for oxysterol metabolites of cholesterol. This feed-forward regulatory loop provides a mechanism for the elimination of excess cholesterol from the body. In this report, we demonstrate that in primary cultures of human hepatocytes, activation of LXRalpha has the opposite effect, repressing CYP7A1 expression. This repression is mediated, at least in part, through induction of the orphan nuclear receptor, short heterodimer partner (SHP), which is also induced by bile acids. We demonstrate that SHP is regulated directly by LXRalpha through a DNA response element that overlaps with the previously characterized bile acid response element. Our data reveal a fundamental difference in the regulation of CYP7A1 in rodent and human hepatocytes and provide evidence that different species employ distinct molecular strategies to regulate cholesterol homeostasis. PMID- 12554796 TI - Transactivation of the progesterone receptor gene in granulosa cells: evidence that Sp1/Sp3 binding sites in the proximal promoter play a key role in luteinizing hormone inducibility. AB - LH induction of the progesterone receptor (PR) in granulosa cells is a central event in ovulation. To identify critical regions of the mouse PR promoter that confer LH inducibility in granulosa cells, a mouse PR promoter (-384/+680) genomic fragment was ligated to a luciferase reporter construct and transfected into primary cultures of granulosa cells. Forskolin/phorbol myristate (PMA) induced PR promoter-luciferase reporter activity in granulosa cells greater than 15-fold. A deletion construct comprised only of the distal promoter alone ( 348/+64) was inactive. Conversely, deletion constructs eliminating putative distal promoter-regulatory elements that bind Sp1, nuclear factor Y, and GATA-4 as well as the transcription start site (+1) failed to reduce forskolin/PMA activation of reporter activity. Additional 5'-deletions identified a minimal promoter region (+420/+680) sufficient to bestow cAMP responsiveness approximately 8- to 10-fold. Two GC-rich regions Sp1(A)[+440/+461] and Sp1(B) [+473/+490] bound Sp1/Sp3. Site-directed mutagenesis of Sp1(A) and Sp1(B) reduced activity of the proximal (+357/+680) promoter reporter construct approximately 50% and 99%, respectively. When the same Sp1(B) mutation was introduced into the intact promoter (-145/+680), forskolin/PMA induction of promoter activity was reduced by 70-80%. When the distal GC box as well as the proximal Sp1(B) site were both mutated in the context of the intact promoter, inducibility of the transgene was even more severely reduced. The importance of these Sp1/Sp3 binding regions was confirmed in human MCF-7 cells and Drosophila SL2 cells. Collectively, these results indicate that the Sp1/Sp3 binding sites within the mouse PR proximal promoter are essential for transactivation of the gene by agonists in granulosa cells. The molecular mechanisms by which LH activates Sp1/Sp3 at this region within the PR gene remain unknown but do not involve changes in the binding of Sp1/Sp3 to the critical GC boxes. Rather, Sp1/Sp3 appear to recruit other factors to the promoter. PMID- 12554797 TI - The many faces of macrophage activation. PMID- 12554798 TI - Interleukin-18. AB - Interleukin-18 (IL-18), a recently described member of the IL-1 cytokine superfamily, is now recognized as an important regulator of innate and acquired immune responses. IL-18 is expressed at sites of chronic inflammation, in autoimmune diseases, in a variety of cancers, and in the context of numerous infectious diseases. This short review will describe the basic biology of IL-18 and thereafter address its potential effector and regulatory role in several human disease states including autoimmunity and infection. IL-18, previously known as interferon-gamma (IFN-gamma)-inducing factor, was identified as an endotoxin-induced serum factor that stimulated IFN-gamma production by murine splenocytes [(1) ]. IL-18 was cloned from a murine liver cell cDNA library generated from animals primed with heat-killed Propionibacterium acnes and subsequently challenged with lipopolysaccharide [(2) ]. Nucleotide sequencing of murine IL-18 predicted a precursor polypeptide of 192 amino acids lacking a conventional signal peptide and a mature protein of 157 amino acids. Subsequent cloning of human IL-18 cDNA revealed 65% homology with murine IL-18 [(3) ] and showed that both contain an unusual leader sequence consisting of 35 amino acids at their N terminus. PMID- 12554799 TI - Dysregulation of transcriptions in primary granule constituents during myeloid proliferation and differentiation in patients with severe congenital neutropenia. AB - We examined the expression of granule constituent genes in myeloid progenitor cells during proliferation and differentiation in patients with severe congenital neutropenia (SCN). The heterozygous mutation of the neutrophil elastase gene was identified in two of four patients. The CD34+/granulocyte-colony stimulating factor receptor (G-CSFR)+ cells of SCN patients showed defective responsiveness to G-CSF in serum-deprived culture. The CD34+/G-CSFR+ cells expressed low levels of the granule constituent mRNAs. The transcription levels of primary granule enzyme genes in CD34+/G-CSFR+ cells were gradually enhanced and then decreased when cells were induced toward myeloid lineage with G-CSF in normal subjects. However, the primary up-regulation and the following down-regulation of these enzyme transcriptions were not clearly observed in SCN patients. No differences in expressions of the lactoferrin gene were seen between normal subjects and patients with SCN. We hypothesize that the abnormal regulation of the transcription in primary granule constituents might involve the defective proliferation and differentiation of myeloid cells in patients with SCN. PMID- 12554800 TI - Activity and subcellular distribution of cathepsins in primary human monocytes. AB - Cathepsins (Cat) in antigen presenting cells (APC) control antigen processing as well as major histocompatibility complex class II transport and function. The set of active Cat and the subcellular architecture of the class II antigen presentation compartment are largely unknown in primary human APC, including peripheral blood monocytes. We used novel chemical tools to visualize Cat in an activity-dependent manner. Primary human monocytes contained active CatS, -B, and -H, while CatL was absent. Expression and activity patterns of Cat in human myelo monocytoid cell lines were distinct from those found in primary cells. On a subcellular scale, the bulk of active Cat was concentrated in lysosomes in primary monocytes. In late endosomes, only active CatS was found in sizable amounts, colocalizing with C-terminal processing of the class II invariant chain and with cystatin C, the major endogenous Cat inhibitor. Late endosomes of human peripheral blood monocytes contain a well-controlled proteolytic machinery distinct from lysosomes, which is likely to play a key role in class II function. PMID- 12554801 TI - Differential recruitment of alpha2beta1 and alpha4beta1 integrins to lipid rafts in Jurkat T lymphocytes exposed to collagen type IV and fibronectin. AB - Collagen type IV (CnIV) and fibronectin (Fn) were used as ligands to study the distribution of alpha(2)beta(1) and alpha(4)beta(1) integrins in low-density, detergent-resistant microdomains (DRM) of Jurkat lymphocytes. CnIV-coated microspheres induced (optical trapping) the redistribution of GM(1)-associated fluorescence from the cell periphery to the area of contact. This was not observed in cells treated with beta-methyl cyclodextrin (MCD). Fn- or bovine serum albumin-coated microspheres did not modify the peripheral distribution of fluorescence. These observations were confirmed by confocal microscopy. Western blot analysis of cells exposed to surfaces coated with CnIV revealed that the alpha(2)-subunit was initially present at low levels in DRM, became strongly associated after 40 min, and returned to basal levels after 75 min. Fn induced a slight recruitment of the beta(1)-integrin alpha(4)-subunit in DRM after 5 and 10 min, followed by a return to basal levels. Neither CnIV nor Fn triggered significant changes in the distribution of the beta(1)-subunit in DRM. Fn- and CnIV-coated microspheres or surfaces coated with these ligands triggered a MCD sensitive mobilization of Ca(2)(+). MCD did not alter the state of the Ca(2)(+) reserves. The differential distributions of the alpha(2)beta(1) and alpha(4)beta(1) integrins in DRM may provide one additional step in the regulation of outside-in signaling involving these integrins. PMID- 12554802 TI - Synergism of nitric oxide and maturation signals on human dendritic cells occurs through a cyclic GMP-dependent pathway. AB - Nitric oxide (NO), generated by phagocytes at inflammation sites, contributes to regulate immune responses through autocrine and paracrine actions on bystander cells. Among the latter are dendritic cells (DCs). Little is known about regulation of DC function by NO, especially in the human system. We exposed human monocyte-derived DCs to the NO donor (z)-1-[2-(2-aminoethyl)-N-(2 ammonioethyl)amino] diazen-1-ium-1,2 diolate (DETA-NO) during their maturation process induced by treatment with tumor necrosis factor alpha or lipopolysaccharide or by CD40 activation. We report here that after exposure to DETA-NO, DCs exhibit a significantly increased ability to activate T lymphocytes stimulated by mycobacterial antigens, Staphylococcus aureus Cowen strain B, allo antigens, or cross-linking of the CD3-T cell receptor complex. This effect persists after removal of DETA-NO, depends on the generation of cyclic guanosine 5'-monophosphate, and is a result of enhanced release by DCs of soluble factors, in particular interleukin (IL)-12. This modulation of DC function is a result of a synergism between NO and the various maturation stimuli, as neither enhanced T cell activation nor IL-12 release was observed after DC exposure to DETA-NO only. These results provide the first evidence that NO acts as a cosignaling molecule regulating human DC response to maturation stimuli. PMID- 12554803 TI - Characterization of promoter elements directing Mona/Gads molecular adapter expression in T and myelomonocytic cells: involvement of the AML-1 transcription factor. AB - Monocytic adaptor (Mona, also called Gads) is a molecular adaptor implicated in T cell activation and macrophage differentiation. The objective of this study was to identify elements regulating specific expression of Mona/Gads in human T cell and myelomonocytic cell lines. We first confirmed that the -2000 to +150 genomic region relative to the Mona gene transcription start site is sufficient to direct specific reporter gene expression in T cell lines, Jurkat, and MOLT-4 and in the immature myeloid cell lines, KG1a and RC2A. Deletion analysis and electrophoresis mobility shift assay identified several cis regulatory elements: overlapping initiator sequences, one interferon response factor-2 (IRF-2)-binding site at position -154, one GC box recognized by Sp1 and Sp3 at position -52, and two acute myeloid leukemia (AML)-1 binding sites at positions -70 and -13. Site directed mutagenesis experiments indicated a key role of AML-1 for driving Mona expression in T cells and myeloid cells, and involvement of Sp1/Sp3 and IRF-2 transcription factors to modulate Mona expression in a cell-specific manner. PMID- 12554805 TI - M-CSF induced differentiation of myeloid precursor cells involves activation of PKC-delta and expression of Pkare. AB - Macrophage-colony stimulating factor (M-CSF) regulates proliferation and differentiation of cells belonging to the monocytic lineage. We investigated the mechanisms of M-CSF differentiation signaling in follicular dendritic cell-P1 cells and analyzed the catalytic activation of different protein kinase C (PKC) isoforms. M-CSF induced rapid catalytic activation of PKC-delta and membrane translocation of the tyrosine phosphorylated form of PKC-delta. Mutation of tyrosine 807 in the M-CSF receptor (Fms) abrogates cell differentiation but not a proliferative response to M-CSF, and FmsY807F failed to activate PKC-delta. We also investigated the downstream signaling pathways from PKC-delta. A cyclic adenosine monophosphate-regulated Ser/Thr kinase gene, protein kinase X (PRKX), has been associated with macrophage differentiation in human cells. We found that M-CSF and PKC-delta induced the expression of the PRKX murine homologue: PKA related gene. Taken together, our results indicate that PKC-delta functions as a critical mediator of M-CSF-induced differentiation signaling. PMID- 12554804 TI - The unique target specificity of a nonpeptide chemokine receptor antagonist: selective blockade of two Th1 chemokine receptors CCR5 and CXCR3. AB - CC chemokine receptor (CCR) 5 and CXC chemokine receptor (CXCR)3 are expressed on T helper cell type 1 cells and have been implicated in their migration to sites of inflammation. Our preceding study demonstrated that a nonpeptide synthetic CCR5 antagonist, TAK-779 (N, N-dimethyl-N-[4-[[[2-(4-methylphenyl)-6, 7-dihydro 5H-benzocyclohepten-8-yl]carbon-yl]amino]benzyl]-tetrahydro-2H-pyran4-aminium chloride, inhibits the development of experimentally induced arthritis by modulating the migration of CCR5(+)/CXCR3(+) T cells to joints. The present study investigated the functional properties of TAK-779, including the effect of this antagonist on CXCR3 function. For this purpose, transfectants expressing mouse CCR5 (mCCR5) or mCXCR3 and expressing mCCR4 or mCXCR4 as controls were established by introducing each relevant gene into 2B4 T cells and were subjected to the following assays. First, the ligand binding to chemokine receptors was assayed by incubating transfectants with [(125)I]-labeled relevant ligand or with the unlabeled relevant ligand followed by staining with anti-ligand antibody. Second, chemokine-induced lymphocyte function-associated antigen-1 (LFA-1) activation was assayed by measuring the adhesion of cells to microculture plates coated with purified intercellular adhesion molecule-1. Third, chemokine stimulated chemotaxis was assayed by observing the cell migration through transwells. In these assays, TAK-779 blocked the ligand binding as well as LFA-1 up-regulating and chemotactic function of mCXCR3 and mCCR5 but did not elicit a biologically significant inhibition of those functions of mCCR4 and mCXCR4. These observations indicate the unique target specificity of TAK-779 and explain why this antagonist efficiently blocks the migration of T cells expressing CCR5 and CXCR3 to sites of inflammation. PMID- 12554806 TI - Telomere shortening in leukocyte subpopulations from baboons. AB - To address questions about telomere length regulation in nonhuman primates, we studied the telomere length in subpopulations of leukocytes from the peripheral blood of baboons aged 0.2-26.5 years. Telomere length in granulocytes, B cells, and subpopulations of T cells all decreased with age. Overall, telomere length kinetics were lineage- and cell subset-specific. T cells showed the most pronounced, overall decline in telomere length. Levels of telomerase in stimulated T cells from old animals were lower than in corresponding cells from young animals. Memory T cells with very short telomeres accumulated in old animals. In contrast, the average telomere length values in B cells remained relatively constant from middle age onward. Individual B cells showed highly variable telomere length, and B cells with very long telomeres were observed after the ages of 1-2 years. In general, cell type-specific telomere kinetics in baboons were remarkably similar to those observed in humans. PMID- 12554807 TI - The improved survival of hematopoietic cells cultured with a fusion protein of insulin-like growth factor II (IGF-II) and interleukin 3 (IL-3) is associated with increases in Bcl-xL and phosphatidylinositol-3 kinase activity. AB - We compared the antiapoptotic activity of a recombinant chimera of insulin-like growth factor II (IGF-II) and interleukin (IL)-3 with the corresponding equimolar mixture of the individual components based on changes in several factors associated with survival in the CD34+ human hematopoietic cell line TF-1. Propidium iodide-stained cells analyzed by fluorescein-activated cell sorter indicated that the chimera was more effective than the corresponding equimolar mixture in decreasing the amounts of apoptotic cells and increasing the proportion of cells in the S-phase of the cell cycle. The chimera was more effective in increasing the antiapoptotic protein Bclx(L) and produced a significant increase in signal transducer and activator of transcription-5 phosphorylation and in phosphatidylinositol-3 kinase (PI-3K) activity. The PI-3K inhibitor LY294002 specifically inhibited cell survival in the presence of the chimera, suggesting a key role of this enzyme in the potentiation of survival caused by the linkage of IGF and IL-3. This potentiation of survival and its preferential inhibition by LY294002 were also observed in a nontransformed, primary culture of human umbilical cord endothelial cells. PMID- 12554808 TI - Differential mRNA expression in circulating gammadelta T lymphocyte subsets defines unique tissue-specific functions. AB - To elucidate the functions of circulating gammadelta T cells, in the absence of antigen stimulation, the differential gene expression of two circulating gammadelta T cell subsets was analyzed. The two subsets, with distinct trafficking phenotypes in young calves, were GD3.5(+), CD8(-), WC1(+) or GD3.5( ), CD2(+), WC1(-), and 90-100% CD8(+) and were sorted based on GD3.5 and gammadelta T cell receptor expression. Results from two different human arrays probed with cDNA from these gammadelta T cell subsets indicated that they have markedly different tissue-specific functions. The genes preferentially expressed by GD3.5(+) (CD8(-)) gammadelta T cells demonstrated that they were highly activated, proliferative, and inflammatory, whereas those expressed by GD3.5(-) (primarily CD8(+)) gammadelta T cells were involved in promoting quiescence, consistent with a role for gammadelta T cells as sentinel mucosal cells, and several were interferon-regulated genes. Gene expression and phenotypic assays indicated that CD8(+) gammadelta T cells were apoptotic, whereas CD8(-) gammadelta T cells were apoptosis-resistant. Differential expression of multiple genes was confirmed in both arrays: That of 14 genes was confirmed by quantitative reverse transcriptase-polymerase chain reaction and that of seven proteins was confirmed by flow cytometry. This novel, genomic analysis of circulating gammadelta T cell subsets, without confounding effects of the tissue microenvironment, offers new insight into the biology and development of neonatal gammadelta T cells. PMID- 12554809 TI - An apoptosis-differentiation program in human polymorphonuclear leukocytes facilitates resolution of inflammation. AB - Human polymorphonuclear leukocytes (PMNs) are an essential part of innate immunity and contribute significantly to inflammation. Although much is understood about the inflammatory response, the molecular basis for termination of inflammation in humans is largely undefined. We used human oligonucleotide microarrays to identify genes differentially regulated during the onset of apoptosis occurring after PMN phagocytosis. Genes encoding proteins that regulate cell metabolism and vesicle trafficking comprised 198 (98 genes induced, 100 genes repressed) of 867 differentially expressed genes. We discovered that complex cellular pathways involving glutathione and thioredoxin detoxification systems, heme catabolism, ubiquitin-proteasome degradation, purine nucleotide metabolism, and nuclear import were regulated at the level of gene expression during the initial stages of PMN apoptosis. Eleven genes encoding key regulators of glycolysis, the hexose monophosphate shunt, the glycerol-phosphate shuttle, and oxidative phosphorylation were induced. Increased levels of cellular reduced glutathione and gamma-glutamyltransferase and glycolytic activity confirmed that several of these metabolic pathways were up-regulated. In contrast, seven genes encoding critical enzymes involved in fatty acid beta-oxidation, which can generate toxic lipid peroxides, were down-regulated. Our results indicate that energy metabolism and oxidative stress-response pathways are gene-regulated during PMN apoptosis. We propose that changes in PMN gene expression leading to programmed cell death are part of an apoptosis-differentiation program, a final stage of transcriptionally regulated PMN maturation that is accelerated significantly by phagocytosis. These findings provide new insight into the molecular events that contribute to the resolution of inflammation in humans. PMID- 12554810 TI - ENRICHD and SADHART: implications for future biobehavioral intervention efforts. PMID- 12554811 TI - Cardiovascular reactivity: status quo and a research agenda for the new millennium. AB - This article introduces a series of articles that assess the present status of the cardiovascular reactivity construct as well as the progress that has been made since a critical review of the reactivity literature by Pickering and Gerin was published in 1990. PMID- 12554812 TI - Cardiovascular reactivity to psychological challenge: conceptual and measurement considerations. AB - OBJECTIVE AND METHODS: This article is a selective review of recent findings bearing on the conceptualization and measurement of cardiovascular reactivity to psychological challenge, with a focus on several issues relevant to the reliability, content validity, construct validity, and criterion validity of these measures. RESULTS AND CONCLUSIONS: With respect to reliability, use of standardized task demands and aggregated scores are associated with enhanced short-term reliability, but the long-term reliability of cardiovascular reactivity has not been sufficiently documented. With respect to content validity, existing evidence suggests that "vascular" or "cardiac" tasks may evoke responses that reflect similar distributions of individual difference, whereas associations between responses to "physical" and "psychological" tasks are modest. The evidence is not clear at present with respect to the importance of including affective or interpersonal stimuli as part of trait reactivity assessments. With respect to construct validity, existing data show that cardiovascular reactivity to psychological challenge is largely independent of standard measures of autonomic function. With respect to criterion validity, recent studies point to a number of methodological limitations that may have restricted our ability to detect lab-to-life generalizability of reactivity measures in the past. Continued progress in understanding and measuring reactivity as an individual difference dimension is essential in helping us to evaluate emerging evidence examining the relationship between reactivity and disease risk. PMID- 12554813 TI - Toward a causal model of cardiovascular responses to stress and the development of cardiovascular disease. AB - OBJECTIVE: Cardiovascular reactivity is hypothesized to mediate the relationship between stress and cardiovascular disease. We describe three considerations that are crucial for a causal model of cardiovascular responses to stress: the need for laboratory-life generalizability, the role of interactions between environmental exposures and individual response predispositions, and the importance of the duration of both stressor exposure and cardiovascular responding. METHODS: We illustrate current understanding of stress-cardiovascular disease relationships with examples from the human and animal psychophysiology, epidemiology, and genetics literature. RESULTS: In a causal model of reactivity, the usefulness of laboratory assessment rests on the assumption that laboratory based cardiovascular reactivity predicts responses in the natural environment. We find only limited generalizability and suggest that cardiovascular responses to stress can be better understood when examined in the natural environment. The interaction of individual response predispositions and stressor exposures contributes to the development and progression of cardiovascular disease; stress disease relationships could therefore be better understood if predispositions and exposures were assessed simultaneously in interactive models. Cardiovascular responses to stress are likely to be most deleterious when responses are prolonged. Responses may vary in their magnitude, frequency, and duration; however, reactivity captures only response magnitude. The assessment of anticipatory and recovery measures, with response magnitude, may therefore lead to a more useful model of the stress-disease relationship. CONCLUSIONS: A causal model of cardiovascular responses to stress should generalize to the real world, assess interactions between individual predispositions and environmental exposures, and focus on sustained pathogenic exposures and responses. PMID- 12554814 TI - Psychophysiological reactivity: mechanisms and pathways to cardiovascular disease. AB - OBJECTIVE: This article examines possible sources of heightened psychophysiological reactivity in relation to risk for hypertension and coronary artery disease. The idea that exaggerated reactions to psychological stress may predict greater risk for future disease has some support in the psychosomatic and behavioral medicine literature. However, the pathways by which exaggerated reactivity could arise in a given person and the implications of different sources of reactivity for potential disease relationships have received little attention. METHODS: This topic is approached through a selective literature review and by means of a neurophysiologically based model of individual differences in physiological reactivity. Temperament characteristics, cognitive processes, neurophysiology, and peripheral physiology are used to indicate three levels that could contribute to exaggerated physiological reactivity. RESULTS: At the top level in the model, activity of the frontal cortex and limbic system establish cognitive-emotional sources of activation that may underlie exaggerated physiological reactivity. In the absence of these influences, large responses may be more likely when exaggerated subcortical response tendencies are present via the hypothalamus or brain stem. Finally, peripheral alterations may account for larger reactions in persons who have otherwise normal emotional and hypothalamic and brainstem response tendencies. Cognitive-emotional and hypothalamic-brainstem sources of altered reactivity may cause or aggravate disease. In contrast, altered peripheral reactivity suggests that a pathophysiologic process may be present, serving as a marker for disease. CONCLUSIONS: These three levels of analysis allow for organization of existing data in the area of cardiovascular reactivity and for planning future studies in a hypothesis-building framework. PMID- 12554815 TI - Cardiovascular reactivity and development of preclinical and clinical disease states. AB - OBJECTIVE: The objective of this review is to evaluate the evidence for the hypothesis that cardiovascular reactivity can predict the development of preclinical (elevated blood pressure, ventricular remodeling, carotid atherosclerosis) and/or clinical cardiovascular disease states. METHODS: A review of the literature was conducted examining prospective studies. RESULTS: Three large epidemiological studies with long-term follow-up periods (20 years or more) have found blood pressure responses to the cold pressor task to be predictive of subsequent essential hypertension in initially normotensive samples. Studies showing less consistent results have tended to use shorter-term follow-up periods. A larger body of literature demonstrates consistent associations between stress-related cardiovascular reactivity and blood pressure elevations in youth over the course of 1 to 6 years; such relationships have not been consistently shown among adult samples. Moderately consistent evidence points to a positive relationship between reactivity and other measures of subclinical disease (increased left ventricular mass and carotid atherosclerosis) among the few prospective studies that have examined these issues to date. A number of additional factors, however, such as baseline levels of disease risk and exposure to psychosocial stress, seem to moderate these relationships. Health status at baseline also seems to moderate the association between reactivity and clinical coronary heart disease in recent reports: two of three existing studies in initially healthy samples show no evidence of a relationship between reactivity and clinical outcomes, whereas three of four studies in samples with preexisting coronary heart disease or essential hypertension show a positive relationship between reactivity and subsequent disease states. CONCLUSIONS: There is reasonable evidence to suggest that cardiovascular reactivity can predict the development of some preclinical states (eg, increased left ventricular mass and blood pressure) states and perhaps even new clinical events in some patients with essential hypertension or coronary heart disease. However, much more information is needed concerning moderating and potentially confounding variables before the robustness of the positive relationships can become clinically useful. PMID- 12554817 TI - Sleep and mortality. PMID- 12554816 TI - Healthy older adults' sleep predicts all-cause mortality at 4 to 19 years of follow-up. AB - OBJECTIVE: Evidence concerning whether sleep disturbances in older adults predict mortality is mixed. However, data are limited to self-reported sleep problems and may be confounded with other comorbidities. We examined whether electroencephalographic (EEG) sleep parameters predicted survival time independently of known predictors of all-cause mortality. METHODS: A total of 185 healthy older adults, primarily in their 60s through 80s, with no history of mental illness, sleep complaints, or current cognitive impairment, were enrolled in one of eight research protocols between October 1981 and February 1997 that included EEG sleep assessments. At follow-up (mean [SD] = 12.8 [3.7] years after baseline, range = 4.1-19.5), 66 individuals were positively ascertained as deceased and 118 remained alive (total N = 184). RESULTS: Controlling for age, gender, and baseline medical burden, individuals with baseline sleep latencies greater than 30 minutes were at 2.14 times greater risk of death (p =.005, 95% CI = 1.25-3.66). Those with sleep efficiency less than 80% were at 1.93 times greater risk (p =.014, CI = 1.14-3.25). Individuals with rapid eye movement (REM) sleep percentages in the lowest 15% or highest 15% of the total sample's distribution (percentage of REM <16.1 or >25.7) were at 1.71 times greater risk (p =.045, CI = 1.01-2.91). Percentage of slow-wave sleep was associated with time to death at the bivariate level, but not after controlling for potential confounders. CONCLUSIONS: Older adults with specific EEG sleep characteristics have an excess risk of dying beyond that associated with age, gender, or medical burden. The findings suggest that interventions to optimize and protect older adults' sleep initiation, continuity, and quality may be warranted. PMID- 12554818 TI - Disordered sleep, nocturnal cytokines, and immunity in alcoholics. AB - OBJECTIVE: Alcoholics who are at risk for infectious disease show profound disturbances of sleep along with decrements of cellular immunity. This study examined the relationships between sleep, nocturnal expression of immunoregulatory cytokines, and natural killer (NK) cell activity in alcoholic patients as compared with control subjects. METHODS: Alcoholic patients (N = 24) and comparison control subjects (N = 23) underwent all-night polysomnography and serial blood sampling at 23:00, 03:00, and 06:30 hours. Stimulated expression of T(H)1 (interferon gamma, IFN-gamma), anti-inflammatory/T(H)2 (interleukin 10, IL 10), and proinflammatory cytokines (IL-6) was measured along with NK cell activity across the night. RESULTS: Alcoholic patients showed lower levels of IL 6 production, suppression of the IL-6/IL-10 ratio, and a reduction of NK cell activity, coupled with losses of delta sleep and increases of rapid eye movement sleep, as compared with control subjects. In addition, alcoholics showed a persistent low ratio of IFN-gamma/IL-10 and reduced levels of NK cell activity, whereas controls had increases of these two immune measures across the night. IL 6 also differentially changed in the two groups; alcoholics showed increases and controls had decreases of IL-6 from 03:00 hours to 06:30 hours. At 06:30 hours, rapid eye movement sleep predicted increases of IL-6 and decreases of NK cell activity independent of the relative contribution of age and chronic alcohol consumption. At 23:00 hours before sleep onset, levels of IL-10 predicted subsequent amounts of delta sleep. CONCLUSIONS: These data further implicate sleep in the regulation of immune function and suggest that disordered sleep contributes to immune alterations in patients with chronic alcoholism. Moreover, the association between awake levels of the anti-inflammatory/T(H)2 cytokine IL 10 and subsequent amounts of delta sleep support the notion of a bidirectional interplay between cytokines and sleep in humans. PMID- 12554819 TI - Association between low plasma levels of cholesterol and relapse in cocaine addicts. AB - OBJECTIVE: In light of recent studies suggesting the existence of associations between low concentrations of cholesterol and various psychiatric disorders, we decided to explore relationships between cholesterol levels and relapse rates in a group of cocaine addicts who had undergone inpatient detoxification. METHODS: The total cholesterol levels of 38 non-opiate-dependent and non-alcohol-dependent cocaine addicts were determined while they were hospitalized. Drug use was subsequently assessed 3, 6, and 12 months after patients were discharged from the hospital. RESULTS: Comparisons of the cholesterol levels (obtained during hospitalization) of relapsers and nonrelapsers by analyses of covariance with age and weight as covariates revealed significantly lower cholesterol values in patients who relapsed at 3 months (p =.046), 6 months (p =.030), and 12 months (p =.019) after discharge. CONCLUSIONS: This study showed an association between a low total cholesterol level and relapse rates in detoxified cocaine addicts. Reasons for the predictive value of low cholesterol levels for relapse for up to 1 year after cholesterol measurements were made are unclear. These data are preliminary and in need of replication. PMID- 12554820 TI - Self-reported depressive symptoms and stress levels in healthy young men: associations with the cortisol response to awakening. AB - OBJECTIVE: There is evidence that clinical depression and negative mood are associated with elevated basal cortisol levels. Recently, measuring the cortisol response during the first hour in the morning with strict reference to the time of awakening was established as a reliable marker of individual adrenocortical activity. In studies using this marker, a relationship with self-reported stress levels and psychosomatic symptoms has been found. The goal of the present study was to investigate the association of self-reported depressive symptomatology with early morning free cortisol levels and their relationship to measures of stress. METHODS: We assessed the severity of depressive symptoms using the Hamilton Depression Inventory and chronic and acute stress perception in 40 healthy young men. Once a week, for 4 consecutive weeks, subjects provided saliva samples collected at 0, 30, and 60 minutes after awakening. RESULTS: Higher levels of depressive symptomatology were associated with a greater cortisol response after awakening. This association seemed to be stronger when only subjects in the nonclinical range of depression were included. Furthermore, cortisol levels and depressive symptomatology were significantly positively correlated with measures of chronic and acute stress perception. CONCLUSIONS: The present study extends earlier findings of hypothalamus-pituitary-adrenal axis hyperactivity in clinical depression to healthy young men with mild levels of depressive symptomatology. Measuring the cortisol response to awakening is proposed as an economical alternative to traditional approaches for determining basal hypothalamus-pituitary-adrenal axis activity. Associations between depressive symptomatology and chronic stress, as well as implications for future studies, are discussed. PMID- 12554821 TI - Anger expression and risk of stroke and coronary heart disease among male health professionals. AB - OBJECTIVE: Anger expression is a dimension of anger that may be strongly related to coronary heart disease and stroke. To date few cohort studies have evaluated the role of anger coping style in the development of cardiovascular disease. This study prospectively examined the effects of anger expression on incidence of cardiovascular disease. METHODS: Participants were male health professionals (N = 23,522), aged 50 to 85 years old and without previous cardiovascular disease, who responded to a mailed questionnaire incorporating the Spielberger Anger-Out Expression Scale in 1996. The cohort was followed for 2 years (1996-1998). RESULTS: Men with moderate levels of anger expression had a reduced risk of nonfatal myocardial infarction compared with those with lower levels of expression (relative risk: 0.56; 95% confidence interval: 0.32-0.97), controlling for coronary risk factors, health behaviors, use of psychotropic medication, employment status, and social integration. Anger expression was also inversely associated with risk of stroke. The multivariate relative risk of stroke was 0.42 (95% confidence interval: 0.20-0.88), comparing men with higher anger-out scores to men with lower scores. A protective dose-response relationship was observed between anger-out score and risk of stroke (p for multivariate trend test: 0.04). CONCLUSIONS: Among this cohort of older men with high socioeconomic status and relatively low level of anger expression on average, moderate anger expression seemed to be protective against cardiovascular disease over a limited follow-up period. PMID- 12554822 TI - Presurgical depression predicts medical morbidity 6 months after coronary artery bypass graft surgery. AB - OBJECTIVE: Depression has been related to poor medical prognosis in patients with coronary artery disease and to diminished quality of life after coronary artery bypass graft surgery (CABG). However, prior studies have not fully examined the impact of depression on medical outcomes after CABG. The purpose of this study was to determine the independent contribution of presurgical depression to short term medical outcome after CABG. METHODS: Medical, surgical, and psychosocial risk factors were assessed before surgery in 89 male veterans undergoing CABG. In addition, patients completed the Beck Depression Inventory. Medical, surgical, and psychological/quality-of-life outcomes were determined at 6 months of follow up by telephone interview and review of medical records. RESULTS: Of the 89 patients studied, 25 scored 10 or greater on the Beck Depression Inventory. The study population was dichotomized on the basis of this cutoff point. A medical prediction model was developed for each outcome of interest, based on the range of medical, surgical, and psychosocial risk indices assessed. The dichotomized depression index was added to these prediction models as a final step. This depression index was found to independently predict cardiac hospitalizations at 6 months (chi(2) = 4.24, p <.04), continued surgical pain at 6 months (chi(2) = 6.36, p <.01), and failure to return to previous activity at 6 months (chi(2) = 15.04, p <.0001). Presurgical depression also predicted depressed affect at 6 months (chi(2) = 13.16, p <.0003). CONCLUSIONS: Depression is an important independent contributor to medical and psychosocial morbidity up to 6 months after CABG. These findings warrant replication with larger and more diverse populations. PMID- 12554823 TI - Prevalence of depression in hospitalized patients with congestive heart failure. AB - OBJECTIVE: Prevalence estimates of depression in hospitalized patients with congestive heart failure (CHF) differ considerably across studies. This article reports the prevalence of depression in a larger sample of hospitalized patients with CHF and identifies demographic, medical, psychosocial, and methodological factors that may affect prevalence estimates. METHODS: A modified version of the Diagnostic Interview Schedule was administered to a series of 682 hospitalized patients with CHF to determine the prevalence of DSM-IV major and minor depression; 613 patients also completed the Beck Depression Inventory. Medical, demographic, and social data were obtained from hospital chart review, echocardiography, and patient interview. RESULTS: In the sample as a whole, 20% of the patients met the DSM-IV criteria for a current major depressive episode, 16% for a minor depressive episode, and 51% scored above the cutoff for depression on the Beck Depression Inventory (>or=10). However, the prevalence of major depression differed significantly between strata defined by the functional severity of heart failure, age, gender, employment status, dependence in activities of daily living, and past history of major depression. For example, the prevalence ranged from as low as 8% among patients in New York Heart Association class I failure to as high as 40% among patients in class IV. CONCLUSIONS: The prevalence of depression in hospitalized patients with CHF is similar to rates found in post-myocardial infarction patients. However, it is considerably higher in certain subgroups, such as patients with class III or IV heart failure. Further research is needed on the prognostic importance and treatment of comorbid depression in CHF. PMID- 12554824 TI - Single-photon emission computerized tomography and neurocognitive function in patients with chronic fatigue syndrome. AB - OBJECTIVE: The purposes of this study were to compare functional imaging under control and experimental conditions among patients with chronic fatigue syndrome (CFS) and healthy persons and to examine perceived and objective performance on a test of attention and working memory previously found to be difficult for persons with CFS. METHODS: Single-photon emission computerized tomography scans were completed on 15 subjects with CFS and 15 healthy persons twice: at rest and when performing the Paced Auditory Serial Addition Test (PASAT). RESULTS: No group differences were found for performance on the PASAT despite CFS subjects' perceptions of exerting more mental effort to perform the task than healthy subjects. Inspection of the aggregate scans by group and task suggested a pattern of diffuse regional cerebral blood flow among subjects with CFS in comparison with the more focal pattern of regional cerebral blood flow seen among healthy subjects. Between-group region-of-interest analysis revealed that although CFS subjects showed less perfusion in the anterior cingulate region, the change in CFS subjects' activation of the left anterior cingulate region during the PASAT was greater than that observed for healthy subjects. The differences were not attributable to lesser effort by the subjects with CFS, confounding effects of mood perturbation, or to poorer performance on the experimental task. CONCLUSIONS: Further research regarding CFS subjects' diffuse cerebral perfusion and its relationship to inefficient neuropsychological performance is warranted. PMID- 12554825 TI - Influence of socioeconomic status and job control on plasma fibrinogen responses to acute mental stress. AB - OBJECTIVE: An elevation in plasma fibrinogen may be one of the pathways through which low socioeconomic status increases cardiovascular disease risk. This study assessed the influence of socioeconomic status, job control, and social isolation on fibrinogen responses to acute stress. METHODS: The study was conducted with 125 white men and 96 white women aged 47 to 58 years, drawn from the Whitehall II cohort. Socioeconomic status was indexed by grade of employment, with 82 high, 75 intermediate, and 64 low grade participants. Plasma fibrinogen and hematocrit were assessed at baseline, immediately after performance of color-word and mirror tracing tasks, and 45 minutes later. RESULTS: Plasma fibrinogen increased from baseline to stress (from 2.85 +/- 0.57 to 2.92 +/- 0.58 g/liter), remaining elevated 45 minutes after stress (2.89 +/- 0.58 g/liter, p <.001). Fibrinogen concentration was greater in the low than in the high or intermediate employment grade groups, independently of sex, age, body mass index, smoking status, and hematocrit. Fibrinogen responses to acute stress did not differ across employment grades. Women had higher fibrinogen levels than men, but this pattern was abolished in women taking hormone replacement therapy. Men experiencing low job control showed greater fibrinogen responses to acute stress than did those with high job control (p =.003). Fibrinogen levels were greater in socially isolated individuals, but social isolation did not affect responses to acute stress. CONCLUSIONS: Socioeconomic status and acute stress had independent effects on the plasma fibrinogen level. Low job control may influence cardiovascular disease risk in men partly through provoking greater fibrinogen stress responses. PMID- 12554826 TI - Association of negative life event stress with coagulation activity in elderly Alzheimer caregivers. AB - OBJECTIVE: Part of the excess coronary disease rate and overall mortality reported with caregiving stress could relate to a hypercoagulable state. Physiological responses to the chronic distress of caregiving may involve hyperactive sympathetic nervous system responses to superimposed stressors. We wondered whether negative life stress might affect hemostatic function in caregivers. METHODS: The three procoagulant measures thrombin-antithrombin III complex (TAT), fibrin D-dimer (DD), and von Willebrand factor (vWF) antigen were assessed in 54 spousal Alzheimer caregivers (mean age, 73 +/- 6 years). Subjects completed a semistructured interview (Psychiatric Epidemiologic Research Interview, PERI) that assessed for non-caregiving-related life stress (ie, number of negative life events) over the 4-week period before blood sampling. RESULTS: Caregivers reported an average of 2.6 negative life events (range, 0-6). The number of negative life events showed a direct association with plasma DD levels (p =.010). In multiple regression analyses, body mass index (BMI) and life stress were the only significant predictors of DD. Together, BMI, hypertension status, gender, and depression accounted for 23% of the variance in DD (p =.022). After these variables had been controlled for, life stress explained an additional 9% (p =.021) of the variance in DD. On the other hand, the number of life-events did not significantly predict TAT or vWF. CONCLUSIONS: The findings suggest that superimposed life stress on top of the chronic stress of Alzheimer caregiving may elicit a hypercoagulable state that could contribute to coronary disease and to increased overall mortality in this population. PMID- 12554827 TI - Alexithymia and sense of coherence in patients with total spinal cord transection. AB - OBJECTIVE: The study investigated the possibility that total spinal cord transection leading to tetraplegia would affect the ability to experience and identify emotions. It also examined whether the dispositional orientation of "sense of coherence" contributed to self-rated quality of life after spinal cord transection. METHODS: Twenty patients with total spinal cord transection at the level of the sixth cervical vertebrae and 20 age- and sex-matched healthy control subjects completed measures of alexithymia, sense of coherence, and quality of life. RESULTS: There were no differences between the two groups on alexithymia scores. However, spinal injury patients reported significantly decreased quality of life relative to matched healthy control subjects. A strong sense of coherence was associated with better self-reported quality of life. This relationship remained after controlling for current affective status. CONCLUSIONS: We conclude that 1) loss of afferent feedback to the brain via the spinal cord does not have a significant effect on alexithymia scores, particularly factor 1 (difficulty in identifying feelings), and 2) sense of coherence may be an important factor in determining psychological adjustment after serious injury. PMID- 12554828 TI - Adherence to antipsychotic and nonpsychiatric medications in middle-aged and older patients with psychotic disorders. AB - OBJECTIVE: The prevalence and consequences of nonadherence to antipsychotic medications in schizophrenia and related psychotic disorders have been well described; however, little is known about adherence to medications for nonpsychiatric conditions in patients with psychotic disorders. We wished to determine medication adherence in nondemented middle-aged and older Veterans Affairs outpatients with schizophrenia or other psychotic disorders who had been prescribed oral medications for hypertension, hyperlipidemia, or diabetes. METHODS: Medication adherence was assessed by review of medication fill records for 76 patients aged 40 years and older who had been prescribed an oral antipsychotic in addition to an oral agent for hypertension (N = 60), hyperlipidemia (N = 28), or diabetes (N = 24). Up to 12 months of therapy was reviewed, and a compliant fill rate (the number of adherent fills in proportion to the total number of prescription fills) and cumulative mean gap ratio (the number of days when medication was unavailable in relation to the total number of days) were calculated for each medication. RESULTS: The 12-month mean compliant fill rates for antipsychotics, antihypertensives, antihyperlipidemics, and antidiabetics ranged from 52% to 64%. Nonpsychiatric medication adherence rates were similar in patients on typical vs. atypical antipsychotics and did not correlate significantly with antipsychotic adherence rates. CONCLUSIONS: Nonadherence rates were found to be equally problematic for both antipsychotic and nonpsychiatric medications in middle-aged and older patients with psychotic disorders. Interventions to improve adherence to both antipsychotic and nonpsychiatric medications are needed. PMID- 12554829 TI - Structure and allosteric regulation of the alpha X beta 2 integrin I domain. AB - The integrin alpha X beta 2 (CD11c/CD18, p150,95) binds ligands through the I domain of the alpha X subunit. Ligands include the complement factor fragment iC3b, a key component in the innate immune defense, which, together with the expression of alpha X beta 2 on dendritic cells and on other leukocytes, suggests a role in the immune response. We now report the structure of the alpha X I domain resolved at 1.65 A by x-ray crystallography. To analyze structural requirements for ligand binding we made a mutation in the alpha X I domain C terminal helix, which increased the affinity for iC3b approximately 200-fold to 2.4 microM compared with the wild-type domain affinity of approximately 400 microM. Gel permeation chromatography supported a conformational change between the wild-type and mutated domains. Conservation of allosteric regulation in the alpha X I domain points to the functional importance of this phenomenon. PMID- 12554830 TI - Associative memory Hamiltonians for structure prediction without homology: alpha/beta proteins. AB - We describe a method for predicting the structure of alpha beta class proteins in the absence of information from homologous structures. The method is based on an associative memory model for short to intermediate range in sequence contacts and a contact potential for long range in sequence contacts. The coefficients in the energy function are chosen to maximize the ratio of the folding temperature to the glass transition temperature. We use the resulting optimized model to predict the structure of three alpha beta protein domains ranging in length from 81 to 115 residues. The resulting predictions align with low rms deviations to large portions of the native state. We have also calculated the free energy as a function of similarity to the native state for one of these three domains, and we show that, as expected from the optimization criteria, the free energy surface resembles a rough funnel to the native state. Finally, we briefly demonstrate the effect of roughness in the energy landscape on the dynamics. PMID- 12554831 TI - Hazardous waste landfill sites and congenital anomalies. PMID- 12554832 TI - Winter air pollution and respiratory function. PMID- 12554834 TI - Fluid losses and hydration status of industrial workers under thermal stress working extended shifts. AB - AIMS: To assess whether workers under significant thermal stress necessarily dehydrated during their exposure and whether "involuntary dehydration" was inevitable, as supported by ISO 9866 and other authorities. Other objectives were to quantify sweat rates against recommended occupational limits, to develop a dehydration protocol to assist with managing heat exposures, and to understand the role of meal breaks on extended shifts in terms of fluid replacement. METHODS: A field investigation to examine the fluid consumption, sweat rates, and changes in the hydration state of industrial workers on extended (10, 12, and 12.5 hour) shifts under significant levels of thermal stress (wet bulb globe temperature (WBGT) >28 degrees C) was conducted on 39 male underground miners. Urinary specific gravity was measured before, during, and at the completion of the working shift. Environmental conditions were measured hourly during the shift. Fluid replacement was measured during the working periods and during the meal breaks. RESULTS: Average environmental conditions were severe (WBGT 30.9 degrees C (SD 2.0 degrees C), range 25.7-35.2 degrees C). Fluid intake averaged 0.8 l/h during exposure (SD 0.3 l/h, range 0.3-1.5 l/h). Average urinary specific gravity at start, mid, and end of shift was 1.0251, 1.0248, and 1.0254 respectively; the differences between start and mid shift, mid and end shift, and start and end shift were not significant. However, a majority of workers were coming to work in a moderately hypohydrated state (average urinary specific gravity 1.024 (SD 0.0059)). A combined dehydration and heat illness protocol was developed. Urinary specific gravity limits of 1.022 for start of shift and 1.030 for end of shift were selected; workers exceeding these values were not allowed into the workplace (if the start of shift limit was exceeded) or were retested prior to their next working shift (if the end of shift limit was exceeded). A target of 1.015 as a euhydrated state for start of shift was adopted for workforce education. CONCLUSIONS: This study found that "involuntary dehydration" did not occur in well informed workers, which has implications for heat stress standards that do not make provision for full fluid replacement during heat exposure. Fluid replacement during meal breaks was not significantly increased above fluid replacement rates during work time, with implications for the duration and spacing of meal breaks on long shifts. Testing of urinary specific gravity was found to be a good indication of hydration status and a practical method of improving workforce awareness and understanding of this important risk factor. Approximately 10 000 dehydration tests have been conducted under the dehydration protocol in a workforce of 2000 persons exposed to thermal stress and has proved practical and reliable. PMID- 12554833 TI - Acute effects of winter air pollution on respiratory function in schoolchildren in southern England. AB - AIM: To investigate the acute health effects of winter outdoor air pollution (nitrogen dioxide (NO(2)), ozone (O(3)), sulphur dioxide (SO(2)), sulphate (SO(4)(2-)),and particles (PM(10))) on schoolchildren in an area of southern England where levels of SO(2) had been reported to be high. METHODS: A total of 179 children, aged 7-13, from three schools (two urban and one rural location), were included. Peak expiratory flow rate (PEFR) and presence or absence of upper respiratory infections were recorded on 63 school days from 1 November 1996 to 14 February 1997. Air pollution and meteorological data were taken from monitors at each school site. The analysis regressed daily PEFR on pollutant level adjusting for confounders and serial correlation and calculated a weighted pooled estimate of effect overall for each pollutant. In addition, large decrements in PEFR were analysed as a binary outcome. Same day, lag 1, lag 2, and a five day average of pollutant levels were used. RESULTS: There were no clear effects of any pollutant on mean PEFR. In addition, we analysed large PEFR decrements (a binary outcome), observing consistent negative associations with NO(2), SO(4)(2-), and PM(10), although few lag/pollutant combinations were significant: odds ratios (95% CI) for five day average effect: NO(2) 24 h average 1.043 (1.000 to 1.089), SO(4)(2-) 1.090 (0.898 to 1.322), PM(10) 1.037 (0.992 to 1.084). The observed effects of PM(10) (only) were stronger in wheezy children (1.114 (1.057 to 1.174)). There were no consistent negative associations between large decrements and ozone or SO(2). CONCLUSIONS: There is no strong evidence for acute effects of winter outdoor air pollution on mean PEFR overall in this area, but there is evidence for negative effects on large PEFR decrements. PMID- 12554836 TI - Exposure related mutagens in urine of rubber workers associated with inhalable particulate and dermal exposure. AB - AIMS: To determine the relation of the inhalation and dermal exposure routes and mutagenic activity in the urine of rubber workers (n = 105). METHODS: Mutagenic activity of ambient total suspended particulate matter (TSPM), surface contamination wipes, and Sunday and weekday urine samples was assessed with S typhimurium YG1041 in the presence of a metabolic activation system. Each subject was grouped into one of two exposure categories for dermal exposure (high (>/=25 revertants/cm(2)), low (<25 revertants/cm(2))) based on the mutagenic activity detected on likely skin contact surfaces and into two airborne mutagenic exposure categories (high (>/=210 revertants/m(3)), low (<210 revertants/m(3))). The potential influence of skin aberrations and acetylation status (NAT2) on urinary mutagenicity levels was also evaluated. RESULTS: A non-significant increase of +1605 revertants/g creatinine in urinary mutagenicity during the workweek relative to levels observed on Sunday was observed for the total population. Subsequent multivariate regression analyses, with the subjects' weekday urinary mutagenicity levels as the dependent variable, revealed associations with environmental and mainstream tobacco smoke exposure, with the level of mutagenic contamination on surfaces with which the subjects had likely contact, with the subjects' inhalable particulate exposure level, with observed mild skin aberrations, and when the subjects had a slow acetylation phenotype. Similar associations, although weaker were observed with Sunday urinary mutagenicity levels as well, except for the association with slow acetylation phenotype. Based on measured exposure levels it could be estimated that a high potential for exposure to surface contamination with mutagenic activity increased weekday urinary mutagenicity by about 62% when compared to low exposed workers, while high inhalable particulate exposure levels increased weekday urinary mutagenicity levels by about 21%. Subjects with mild skin aberrations had an additional, non significant, increase in weekday urinary mutagenic activity compared to subjects without any skin aberrations. DISCUSSION: Results suggest that the dermal exposure route may contribute more to the level of genotoxic compounds in urine of rubber workers than the inhalation route. Although the study was limited in size, the results warrant further investigation in the importance of and ways to effectively control the dermal exposure route in the rubber industry. PMID- 12554837 TI - Exposure-response relations among laboratory animal workers exposed to rats. AB - AIM: To explore exposure-response relations in a cohort of laboratory animal workers. METHODS: Exposure-response modelling was carried out in a cohort of 342 laboratory animal workers. Three exposure indices, divided into different exposure categories, were used in the analyses: intensity of exposure to rat urinary aeroallergen (RUA, the main allergen workers were exposed to), weekly duration of exposure to rats, and the product of the intensity and weekly duration of exposure. Outcomes studied were work related chest, eyes and nose, and skin symptoms that had started after employment at the sites, specific sensitisation, and a combination of symptoms and sensitisation. Cox proportional hazard modelling was used to explore exposure-response relations. Smoking, atopic status, age, and gender were taken into account. RESULTS: We observed the clearest exposure-response relations for the intensity of exposure to RUA and the various endpoints. No clear exposure-response relations were observed for the weekly duration of exposure or the product of the intensity and weekly duration of exposure. The strongest and clearest exposure-response relations for symptoms were observed among rat sensitised workers, while the non-sensitised workers only showed small increased risks of developing symptoms without clear exposure response relations. Sensitised workers were almost four times more likely to go on to develop chest symptoms compared to non-sensitised workers. PMID- 12554838 TI - Ambiguous relation between physical workload and low back pain: a twin control study. AB - AIMS: To examine the association between self reported physical workload and low back pain (LBP) in younger twins. To investigate whether genetic factors interact with physical workload in relation to LBP. METHODS: A twin control study was performed within a population based twin register using 1910 complete monozygotic (MZ) and same sexed dizygotic (DZ) twin pairs aged 25-42 and discordant for LBP. LBP in the affected twins was divided into two groups: "LBP for 30 days during the past year". Physical workload was divided into four categories: "sitting", "sitting/walking", "light physical", and "heavy physical". Data were analysed in a matched design using conditional logistic regression. MZ and DZ twins were analysed separately and together in order to determine possible genetic influences in relation to physical workload and LBP. RESULTS: Statistically significant graded relations were found for increasing workload and LBP of longer duration but not for LBP of shorter duration ( or =3 admissions for COPD in the year before recruitment (hazard ratio (HR)=1.66, 95% CI 1.16 to 2.39), forced expiratory volume in 1 second (FEV(1)) percentage predicted (0.97, 95% CI 0.96 to 0.99), oxygen tension (0.88, 95% CI 0.79 to 0.98), higher levels of usual physical activity (0.54, 95% CI 0.34 to 0.86), and taking anticholinergic drugs (1.81, 95% 1.11 to 2.94). Exposure to passive smoking was also related to an increased risk of readmission with COPD after adjustment for clinical factors (1.63, 95% CI 1.04 to 2.57) but did not remain in the final model. CONCLUSIONS: This is the first study to show a strong association between usual physical activity and reduced risk of readmission to hospital with COPD, which is potentially relevant for rehabilitation and other therapeutic strategies. PMID- 12554889 TI - Imbalance between levels of nitrogen oxides and peroxynitrite inhibitory activity in chronic obstructive pulmonary disease. AB - BACKGROUND: The prevalent theory concerning the pathogenesis of chronic obstructive pulmonary disease (COPD) is of an imbalance between oxidants and antioxidants in the lung. It has recently been reported that the production of peroxynitrite, an extremely potent oxidant, is increased in the airways of patients with COPD. A study was undertaken of the imbalance between the levels of nitrogen oxides and antioxidant activity against peroxynitrite in the airways of patients with COPD. METHODS: Sputum induction was performed in 30 patients with COPD and 15 normal control subjects. Levels of nitrogen oxides, percentage of neutrophils, and interleukin 8 (IL-8) levels were measured in sputum samples, and peroxynitrite inhibitory activity was assayed by monitoring rhodamine formation. RESULTS: Nitrite and nitrate levels in induced sputum were significantly higher in patients with COPD than in normal controls (949 (133) microM v 621 (89) microM, p<0.001). In contrast, peroxynitrite inhibitory activity in induced sputum was significantly lower in patients with COPD than in normal controls (47.4 (12.7)% v 92.9 (3.9)%, p<0.001). There was a negative correlation between nitrite and nitrate levels and peroxynitrite inhibitory activity in induced sputum (r=-0.775, p<0.001). Peroxynitrite inhibitory activity was also significantly correlated with forced expiratory volume in 1 second (FEV(1)) % predicted (r=0.539, p=0.004), FEV(1)/FVC (r=0.512, p=0.006), and carbon monoxide transfer factor (TLCO) (r=0.486, p=0.009). Moreover, there was a significant negative correlation between peroxynitrite inhibitory activity and the degree of neutrophilic inflammation (percentage of neutrophils: r=-0.754, p<0.001; IL-8 levels: r=-0.497, p=0.007). CONCLUSIONS: Reduced peroxynitrite inhibitory activity and increased levels of nitrogen oxides are found in induced sputum from patients with COPD. An imbalance in nitrogen oxides and antioxidant defence may contribute to the pathogenesis of COPD. PMID- 12554890 TI - Breathing retraining for dysfunctional breathing in asthma: a randomised controlled trial. AB - BACKGROUND: Functional breathing disorders may complicate asthma and impair quality of life. This study aimed to determine the effectiveness of physiotherapy based breathing retraining for patients treated for asthma in the community who have symptoms suggestive of dysfunctional breathing. METHODS: 33 adult patients aged 17-65 with diagnosed and currently treated asthma and Nijmegen questionnaire scores > or =23 were recruited to a randomised controlled trial comparing short physiotherapy breathing retraining and an asthma nurse education control. The main outcome measures were asthma specific health status (Asthma Quality of Life questionnaire) and Nijmegen questionnaire scores RESULTS: Of the 33 who entered the study, data were available on 31 after 1 month and 28 at 6 months. The median (interquartile range) changes in overall asthma quality of life score at 1 month were 0.6 (0.05-1.12) and 0.09 (-0.25-0.26) for the breathing retraining and education groups, respectively (p=0.018), 0.42 (0.11-1.17) and 0.09 (-0.58-0.5) for the symptoms domain (p=0.042), 0.52 (0.09-1.25) and 0 (-0.45-0.45) for the activities domain (p=0.007), and 0.50 (0-1.50) and -0.25 (-0.75-0.75) for the environment domain (p=0.018). Only the change in the activities domain remained significant at 6 months (0.83 (-0.10-1.71) and -0.05 (-0.74-0.34), p=0.018), although trends to improvement were seen in the overall score (p=0.065), the symptoms domain (p=0.059), and the environment domain (p=0.065). There was a correlation between changes in quality of life scores and Nijmegen questionnaire scores at 1 month and at 6 months. The number needed to treat to produce a clinically important improvement in health status was 1.96 and 3.57 at 1 and 6 months. CONCLUSION: Over half the patients treated for asthma in the community who have symptoms suggestive of dysfunctional breathing show a clinically relevant improvement in quality of life following a brief physiotherapy intervention. This improvement is maintained in over 25% 6 months after the intervention. PMID- 12554891 TI - Relationship between induced sputum eosinophils and the clinical pattern of childhood asthma. AB - BACKGROUND: The relationship between the clinical pattern of asthma and airway inflammation in childhood asthma is poorly characterised, yet underpins the treatment recommendations in current asthma guidelines. A study was undertaken to examine the relationship between airway inflammation and clinical asthma in children. METHODS: Children with asthma (n=146) and healthy controls (C, n=37) were recruited from primary and specialist clinics. Sputum induction and hypertonic saline challenge were performed. RESULTS: As the frequency of asthma episodes in the past 12 months increased, there were significant increases in sputum eosinophils (median; infrequent episodic (IE) 1.5%, frequent episodic (FE) 2.3%, persistent (P) 3.8%, control (C) 1.0%; p=0.002), sputum eosinophil cationic protein (ECP) (IE 113 ng/ml, FE 220, P 375, C 139; p=0.003), and desquamated bronchial epithelial cells (IE 2.0%, FE 6.0%, P 5.0%, C 2.5%; p=0.04). Treatment intensity was also associated with increased sputum eosinophils (p=0.005). The relationships between other severity markers (current symptoms, lung function) were less strong. CONCLUSION: Children with more frequent episodes of clinical asthma exhibit increasing airway inflammation that is characterised by sputum eosinophilia and bronchial epithelial desquamation. The results support clinical assessment by frequency of wheezing episodes over the past 12 months when determining anti-inflammatory treatment requirements, and indicate that current symptoms are determined by mechanisms in addition to sputum eosinophilia. PMID- 12554893 TI - Increased collagenase activity in macrophages from bronchial lavage as a diagnostic marker of non-small cell lung cancer. AB - BACKGROUND: The roles of matrix metalloproteinases (MMPs) in cancer metastasis have been studied. Macrophages are considered to release MMPs in the tissues of patients with lung cancer. METHODS: Intracellular collagenase activity was measured in CD14+ CD45+ cells from bronchial lavage fluid to establish a new diagnostic tool for lung cancer. Between August 2000 and November 2001 bronchoscopy and bronchial lavage were performed in 45 patients with abnormal shadows on the chest radiograph; 21 had lung cancer and 24 had non-malignant disease. RESULTS: Collagenase activity in patients with primary lung cancer (5.54 (0.65)) or non-small cell lung cancer (NSCLC) (5.62 (0.71)) was significantly higher than in those with non-malignant disease (3.63 (0.78), p=0.006 and p=0.008, respectively). Only three of 18 patients in the low activity group were diagnosed as having cancer compared with 18 of 27 in the high activity group (p=0.001). This significance was not seen in non-smokers but it was apparent in smokers/ex-smokers. Excluding non-smokers improved the specificity of collagenase activity in differentiating cancer and non-malignant disease from 62.5% to 80.0%. The sensitivity of the test was 85.7% in all patients and 88.2% in smokers/ex smokers. CONCLUSIONS: Measurement of intracellular collagenase activity in macrophages in bronchial lavage fluid is a useful diagnostic tool for distinguishing between cancer and non-malignant diseases, especially in smokers and ex-smokers. PMID- 12554895 TI - Impaired recycling of surfactant-like liposomes in type II pneumocytes from injured lungs. AB - BACKGROUND: Surfactant synthesis and secretion has been shown to be impaired in type II cells from diseased lungs. The mechanism of surfactant lipid recycling, which is an important physiological process in surfactant treatment, was studied in type II cells isolated from injured lungs. METHODS: Different stages of lung injury were induced by exposing rats to 10 ppm nitrogen dioxide (NO(2)) for 3, 20, and 28 days. Type II cells were isolated from these lungs and recycling of (3)H-DPPC labelled surfactant-like liposomes was studied in vitro. RESULTS: Uptake of liposomes (150 micro g/ml) for 20 minutes in the absence and presence of surfactant protein-A (SP-A, 5 micro g/ml) was higher in cells from NO(2) injured lungs (63-78%) than in control cells. There was no difference in liposome uptake between the groups with NO(2) exposure of different duration. After liposome uptake, most of the internalised label remained in the phosphatidylcholine (PC) fraction and increased with duration of exposure to NO(2). After 20 minutes internalisation, cells were allowed to resecrete lipids for a further 20 minute period. In cells from controls and from all stages of lung injury, liposomes that had been internalised in the presence of SP-A were resecreted to a greater extent than those internalised without SP-A. However, cells from lungs exposed to NO(2) resecreted less lipid than cells from control lungs. Again, there was no difference in resecretion between the groups with NO(2) exposure of different duration. CONCLUSION: Type II cells from injured lungs internalise more surfactant-like liposomes than cells from controls, suggesting a putative therapeutic significance to cope with limited alveolar surfactant pools in lung injury. PMID- 12554896 TI - Relationship between inflammatory cells and structural changes in the lungs of asymptomatic and never smokers: a biopsy study. AB - BACKGROUND: A study was undertaken to investigate the relationship between inflammatory cells and structural changes in the mucosa of the airways in an epidemiological sample of a group of asymptomatic smokers (smokers who had never sought medical attention for respiratory problems) and in non-smoking subjects. METHODS: Bronchial biopsy specimens were taken from 29 smokers and 16 never smokers and stained with monoclonal antibodies HNL, EPO, AA1, CD68 in order to identify neutrophils, eosinophils, mast cells, and macrophages, respectively. The biopsy specimens were also stained with monoclonal antibodies to the cytokines interleukin (IL)-1beta and IL-8. Structural changes were identified by staining the biopsy specimens with antibodies to tenascin and laminin and by evaluating the condition of the epithelial layer. RESULTS: The numbers of all inflammatory cells and of cytokine staining cells were significantly increased in smokers. The thickness of the tenascin and laminin layers was increased in the smoking group and the integrity of the epithelial layer was significantly reduced. In smokers the epithelial integrity was negatively correlated with the number of eosinophils and macrophages. The thickness of the tenascin and laminin layers was positively correlated with AA1 and EPO positive cells only. CONCLUSION: High numbers of inflammatory cells are present in the bronchial mucosa of asymptomatic smokers which have a clear relationship with the impaired epithelial integrity. The increased thickness of the laminin and tenascin layers in these subjects was strongly related to the presence of eosinophils and mast cells, suggesting a role for these cells in tissue remodelling of the airways of smokers. PMID- 12554898 TI - Radiological versus histological diagnosis in UIP and NSIP: survival implications. AB - BACKGROUND: High resolution computed tomography (HRCT) has an important diagnostic role in idiopathic interstitial pneumonia (IIP). We hypothesised that the HRCT appearance would have an impact on survival in patients with IIP. METHODS: HRCT scans from patients with histological usual interstitial pneumonia (UIP; n=73) or histological non-specific interstitial pneumonia (NSIP; n=23) were characterised as definite UIP, probable UIP, indeterminate, probable NSIP, or definite NSIP. Cox regression analysis examined the relationships between histopathological and radiological diagnoses and mortality, controlling for patient age, sex, and smoking status. RESULTS: All 27 patients with definite or probable UIP on HRCT had histological UIP; 18 of 44 patients with probable or definite NSIP on HRCT had histological NSIP. Patients with HRCT diagnosed definite or probable UIP had a shorter survival than those with indeterminate CT (hazards ratio (HR) 2.43, 95% CI 1.06 to 5.58; median survival 2.08 v 5.76 years) or HRCT diagnosed definite or probable NSIP (HR 3.47, 95% CI 1.58 to 7.63; median survival 2.08 v 5.81 years). Patients with histological UIP with no HRCT diagnosis of probable or definite UIP fared better than patients with histological UIP and an HRCT diagnosis of definite or probable UIP (HR 0.49, 95% CI 0.25 to 0.98; median survival 5.76 v 2.08 years) and worse than those with a histological diagnosis of NSIP (HR 5.42, 95% CI 1.25 to 23.5; median survival 5.76 v >9 years). CONCLUSIONS: Patients with a typical HRCT appearance of UIP experience the highest mortality. A surgical lung biopsy is indicated for patients without an HRCT appearance of UIP to differentiate between histological UIP and NSIP. PMID- 12554899 TI - Pigtail drainage in the treatment of tuberculous pleural effusions: a randomised study. AB - BACKGROUND: Tuberculous pleurisy can result in obvious clinical symptoms, pleural fibrosis, and pleural thickening. Some studies of tuberculous pleurisy have suggested that symptomatic improvement and minimisation of sequelae can be achieved by completely draining the effusion during treatment, although the results have not been conclusive. METHODS: Sixty one patients with tuberculous pleurisy were divided into two groups; 30 patients received pigtail drainage combined with antituberculosis (TB) drug treatment and 31 received only anti-TB drugs. Outcome measurements were assessed for a period of 24 weeks after treatment and included symptom scores and the incidence of residual pleural thickening (RPT). RESULTS: Although the duration of dyspnoea was significantly shortened by the use of pigtail drainage (median 4 days (IQR 4-5) v 8 days (IQR 7 16), p<0.001), a comparison of combined mean (SD) visual analogue scale (VAS) scores showed no significant difference between the groups after one week of treatment (57.1 (33.2) v 68.5 (44.7) or at any time during the follow up period. The incidence of RPT of more than 10 mm in the group treated with pigtail drainage and anti-TB drugs was 26% compared with 28% in the group receiving drug treatment only. The incidence of RPT levels of more than 2 mm in the two groups was 50% and 51%, respectively. No statistical difference between the two groups in terms of forced vital capacity was found at the end of treatment (median (IQR) 85.5% (69-94) of predicted v 88% (78-96) of predicted). CONCLUSION: The addition of pigtail drainage to an effective anti-TB regimen is not clinically relevant and does not reduce the level of RPT. PMID- 12554900 TI - Impaired cough reflex in patients with recurrent pneumonia. AB - BACKGROUND: A substantial proportion of patients with recurrent pneumonia do not have an apparent underlying condition, but they may have unknown defects in host defence mechanisms such as cough reflex. METHODS: Capsaicin cough sensitivity was measured in seven patients with recurrent pneumonia but no underlying condition. Recurrent pneumonia was defined as at least two episodes of pneumonia in 1 year, or three or more episodes at any time. After remission of pneumonia, 10 doubling concentrations of capsaicin (1.22-625 microM) were inhaled until five or more coughs were induced (cough threshold). Mucociliary clearance was assessed on the basis of nasal ciliary beat frequency and nasal clearance time. Twenty one age and sex matched healthy subjects were studied as controls. RESULTS: Each patient had 2-6 episodes of pneumonia. Most episodes developed in dependent lung segments, suggesting the involvement of silent aspiration. Log transformed cough threshold was significantly higher in patients than in controls (mean 2.37 M (95% CI 1.84 to 2.90) v 1.29 microM (95% CI 1.11 to 1.47); p<0.0001). The effect of pneumonia per se on cough reflex seemed unlikely since the cough threshold showed little change when re-examined after 3 months. The patients and controls did not differ with respect to the indices of mucociliary clearance. CONCLUSIONS: Impaired cough reflex may be involved in the pathogenesis of recurrent pneumonia. PMID- 12554901 TI - Association of IL-10 polymorphism with severity of illness in community acquired pneumonia. AB - BACKGROUND: The influence of genetic polymorphisms of interleukin (IL)-10, tumour necrosis factor (TNF)-alpha, and IL-6 gene promoters on severity of systemic inflammatory response syndrome (SIRS) associated with community acquired pneumonia (CAP) was studied. METHODS: Using PCR-RFLP analysis we analysed a 1082G/A single nucleotide polymorphism (SNP) of the anti-inflammatory IL-10 gene, a -308G/A SNP of the pro-inflammatory TNF-alpha gene and a -174G/C SNP of the IL 6 gene. Illness severity was stratified according to SIRS score, calculated by presence of up to four physiological indices: temperature, white blood cell count, heart rate and respiratory rate (non-SIRS, SIRS 2, SIRS 3, and SIRS 4). RESULTS: A statistically significant stepwise increase in frequency of the IL-10 G allele, associated with higher expression of the gene, was observed in patients with increasing severity of illness from non-SIRS (n=19) to SIRS 2 (n=17), SIRS 3 (n=33) and SIRS 4 (n=24). This was primarily due to a higher frequency of the GG genotype with increasing severity from non-SIRS through to SIRS 4. IL-10 G allele frequency was also increased in patients who died as a result of CAP (n=11) compared with CAP survivors (n=82) (p=0.01). No association was seen between the TNF-alpha -308G/A and IL-6 -174G/C SNPs and disease. Additionally, no interaction between all three SNP genotypes and disease severity was observed. CONCLUSIONS: A polymorphism affecting IL-10 expression may influence the severity of illness in patients with CAP. PMID- 12554902 TI - The pulmonary physician in critical care. 13: the pulmonary circulation and right ventricular failure in the ITU. AB - The management of severe pulmonary hypertension associated with right ventricular failure is reviewed and its relevance to adults with acute respiratory distress syndrome (ARDS) is discussed. PMID- 12554906 TI - Mediastinal cavernous haemangioma in a patient with Klippel-Trenaunay syndrome. AB - The Klippel-Trenaunay syndrome (KTS) is a rare syndrome characterised by the triad of varicose veins, bony and soft tissue hypertrophy, and cutaneous haemangioma. A 30 year old man with KTS with a right mediastinal mass which progressively enlarged over 5 years is described. Computed tomography, magnetic resonance imaging, and bronchial angiography revealed a vascular lesion in the azygous area. After complete excision of the mass, histological examination revealed cavernous haemangioma. To our knowledge, this is the first report of intrathoracic haemangioma in KTS. PMID- 12554907 TI - Pneumocystis carinii pneumonia with pleurisy, platypnoea and orthodeoxia. AB - We present a patient who collapsed with chest pain and dyspnoea on a transatlantic flight. She was found to have Pneumocystis carinii pneumonia (PCP) and human immunodeficiency virus infection. Platypnoea and orthodeoxia, which have not been previously reported in association with PCP, were major features of her illness. The PCP predominantly affected her lung bases and it is likely that gravity increased intrapulmonary blood flow through poorly ventilated lung bases with failure of pulmonary vasoconstriction to increase upper zone perfusion, exacerbating desaturation on sitting up. The partial DNA sequence of the infecting P carinii was identical to previously described isolates. PMID- 12554909 TI - Ultrastructural examination of bronchial specimens from children with moderate asthma. PMID- 12554905 TI - Multiple roles of nitric oxide in the airways. AB - Nitric oxide is endogenously released in the airways by nitric oxide synthase. Functionally, two isoforms of this enzyme exist: constitutive and inducible. The former seems to protect airways from excessive bronchoconstriction while the latter has a modulatory role in inflammatory disorders of the airways such as asthma. This review explores the physiological and pathophysiological role of endogenous nitric oxide in the airways, and the clinical aspects of monitoring nitric oxide in exhaled air of patients with respiratory disease. PMID- 12554910 TI - Declaring competing interests. PMID- 12554904 TI - Pharmacotherapy and airway remodelling in asthma? AB - Over the last few decades attention has largely focused on airway inflammation in asthma, but more recently it has been appreciated that there are important structural airway changes which have been grouped together under the term "airway remodelling". It is only now that questions have been asked about the impact of treatment on these structural changes. This review examines the nature of these structural airway changes, the mechanisms of their generation, their potential consequences, and what is known about the ability of anti-asthma treatments to modulate these changes. PMID- 12554911 TI - Inappropriate antibiotic prescription to children with acute respiratory infection in Brazil. AB - OBJECTIVE: To determine the rate of inappropriate antibiotic prescription and to describe the types of antibiotics prescribed by health workers to children with acute respiratory infection (ARI). DESIGN: Cross-sectional survey conducted in 6 state capitals of Brazil. METHODS: A representative sample of facilities was selected in each state using a cluster sampling method based on the mean number of visits of children less than 5 years of age. In each facility, consultations were observed and children were reassessed following standard guidelines. Health worker s diagnosis and treatment were compared with a gold standard and inappropriate antibiotic prescriptions noted. RESULTS: 1565 children with ARI from 156 health facilities (73% health centers) were included in study. Most children had a common cold (77.5%). Antibiotics were inappropriately prescribed in 9.2% (95% CI: 7.8, 10.7) of ARI cases (range: 2.8% to 25%). Most frequently prescribed antibiotics were those recommended by the ARI Program. Seventy-six percent of health workers explained to guardians how to use antibiotics at home and 3.9% demonstrated the first dose. Antibiotics were available in 84% of health facilities. CONCLUSION: Inappropriate prescription of antibiotics varied geographically in Brazil. More training and supervision is needed to decrease it. PMID- 12554912 TI - Humanized care of preterm babies. PMID- 12554913 TI - Infantile cystinosis. AB - Ten patients of nephropathic cystinosis were admitted during the period 1995 2000. Their mean age was 12 months. The signs of failure to thrive and advanced rickets were seen in all patients. Other features included polyuria, polydipsia, pathologic fractures and deafness. Laboratory findings included glucosuria, hyposthenuria, hypocalcemia, proteinuria and azotemia. Therapy with phosphocysteamine showed marked clinical improvement. PMID- 12554914 TI - Essential hypertension: antecedents in children. AB - A prospective study was conducted to determine the incidence of essential hypertension (EH) and identify markers, if any, in children of essential hypertension families. The study group included 90 children (2-18 years) with a parent or grandparent with EH while the control group had 25 age matched children from non-hypertensive families. Around 30% children (n=27) from these families had a diastolic blood pressure of >95th centile and an additional 27% (n=24) had borderline hypertension. The serum cholesterol, serum triglycerides and 24 hour urinary sodium excretion were significantly higher in the study group (p < 0.05) as compared to controls. The children from the study group also had a significant high salt (p < 0.001) and fat intake (P < 0.05). PMID- 12554915 TI - McCune Albright Syndrome (MCAS): a case series. AB - McCune Albright Syndrome (MCAS) is an association of, Cafe-au-lait macules, polyostotic fibrous dysplasia and autonomous hyperfunctioning endocrinopathy. This is a rare disorder seen more commonly in females. We evaluated 7 (6F & 1M) cases under six years of age (4 months to 5.5 yrs) presenting with Cafe-au-lait spots, polyostotic fibrous dysplasia and/or sexual precocity. All the 7 cases had large Cafe-au-lait spots, radiologic features of polyostotic fibrous dysplasia were seen in 5 cases. Six girls had precocious puberty with large ovarian follicles and elevated S. Estradiol levels (14-65 pg/dl) with prepubertal gonadotropin levels in 5 of them. Medroxy-progrestrone acetate was used to treat the sexual precocity. Five girls on follow up for 6 months (6mo-16mo) showed cessation of menstrual episodes and regression of ovarian follicles in three, regression in breast size in one, and three girls continued to grow at a height velocity >95th centile for age. Skeletal lesions and skin features did not show any change. No other endocrinopathy was noted. Gonadotropin independent precocious puberty was the only endocrine affection seen in this series. PMID- 12554916 TI - A study of Vitamin K status in children on prolonged antibiotic therapy. AB - Vitamin K deficiency is known to cause coagulopathy and bleeding in patients on prolonged antibiotic therapy. This study was conducted to evaluate the status of vitamin K deficiency in hospitalized children on prolonged antibiotic therapy and its role in reversing the coagulopathy. A prospective non-randomized study was conducted on children on antibiotic therapy at a tertiary care hospital. Children in the 1 month-1 year age group developed significant coagulopathy as compared to other age groups. Coagulation abnormalities were also seen to be more in children with greater grades of malnutrition, on a more prolonged course of antibiotics and in children who were critically ill in intensive care. Hypoprothrombinemia previously reported to be due to B-lactam antibiotics containing the N-Methyl Thio Tetrazole (NMTT) group also resulted from antibiotics without this side chain. Inhibition of intestinal microorganisms by antibiotics was thought to be a likely explanation of this phenomenon. We suggest Vitamin K prophylaxis in severely ill patients, on extended periods of antibiotics and inadequate diet to prevent morbidity and mortality. PMID- 12554917 TI - Medium and long term central venous access in children. AB - From September 2000 to August 2001, 104 central venous access devices (CVAD) were inserted in 91 children, governed by a uniform protocol. Thirty catheters were inserted in neonates, 29 in infants, 37 in children and 8 in adolescents. Fifty one were planned insertions in the operating suite and 53 were emergencies - often by the bedside. There were 12 insertion related complications-all of which were minor. Neonatal age and bedside introduction had a higher risk of insertion related problems. The incidence of non-infectious complications was 20% (rate of 13.7/1000 line days) and was influenced by the child's age and insertion site. Femoral route was the safest. Incidence of catheter associated infections (CAI) was 15.4% (rate of 11/1000 line days). Only 2 children had catheter associated bloodstream infection. Neonates were at higher risk of catheter related infections. Age, insertion site and occurence of insertion complications influenced duration of catheterization (median 7.5 days, range 2-243 days) There was no major complication, though more than 50% insertions were in neonates and infants. In our practice, use of CVAD is feasible and safe, especially in neonates and infants. PMID- 12554918 TI - Psedohypoparathyroidism in newborn - a rare presentation. AB - A 7-day-old male baby was referred with history of persistent multifocal convulsions from second day of life. He was found to have hypocalcemia, hyperphosphatemia, normal serum magnesium and normal renal function tests. Serum parathormone was found to be elevated. Baby was diagnosed as case of pseudohypoparathyroidism and was treated with calcium supplementation and calcitriol. At 9 months he was asymptomatic on treatment, with normal serum calcium and phosphorus. PMID- 12554919 TI - Pseudo tumoral laryngeal tuberculosis. AB - An 11-year-old female child presented with high grade intermittent fever and cough for a duration of 6-7 months and hoarseness of voice for 6 months. Skiagram of the chest showed evidence of miliary mottling. Direct laryngoscopic examination revealed inflammatory swelling over left vocal cord. The biopsy of the swelling showed chronic granulomatous lesion. Patient improved remarkably with anti-tubercular therapy. PMID- 12554920 TI - Trichotillomania. AB - Trichotillomania, though uncommon, is one of the causes of unexplained hair loss, especially in children. Three girls in the age group of 4-6 years were observed in our pediatric dermatology clinic to have trichotillomania. In one child, there was co-existent alopecia areata. All were referred to the child guidance clinic and they all showed improvement with behavior therapy. A close liasion between the dermatologist, psychiatrist and parents would go a long way in preventing this alopecia PMID- 12554921 TI - Hodgkins disease in relapse presenting as bone tumor. AB - A 10-year-old male child presented with multiple lymph node swellings. A diagnosis of Hodgkins disease was made on histopathological examination. The patient developed relapse six months after his last chemotherapy as a solitary bone tumor, which is rare. Immunohistochemical evaluation helped for the correct typing of Hodgkins disease. PMID- 12554922 TI - Chronic bullous dermatosis of childhood. PMID- 12554923 TI - Magnetic Resonance Imaging (MRI) in the diagnosis of fetus-in-fetu. PMID- 12554924 TI - Hyperkalemia and ventricular tachycardia in ELBW infant. PMID- 12554925 TI - Professional appearance of pediatrician: children's and pediatricians opinion. PMID- 12554926 TI - Truncus arteriosus in fetal hydantoin syndrome - a new association? PMID- 12554927 TI - Prevalence of anemia and worm infestation in school going girls at Gulbargha, Karnataka. PMID- 12554928 TI - Neonatal Hypernatremia due to high Breast milk sodium. PMID- 12554930 TI - Hib vaccination controversy and the declaration of commercial interest. PMID- 12554931 TI - Lysosomal cysteine proteases (cathepsins): promising drug targets. AB - Papain-like lysosomal cysteine proteases are processive and digestive enzymes expressed in organisms from bacteria to humans. Their ubiquity alone makes them potential drug targets, with the assumption that appropriate specificities may be achieved. These enzymes have rather short active-site clefts, comprising three well defined substrate-binding subsites (S2, S1 and S1') and additionally have comparatively broad binding areas (S4, S3, S2', S3'). This geometry distinguishes them from other protease classes, such as serine and aspartic proteases, with six and eight substrate-binding sites, respectively. Exopeptidases (cathepsins B, C, H and X), in contrast to endopeptidases (such as cathepsins L, S, V and F), possess structural features that facilitate binding of N- and C-terminal groups of substrates in the active-site cleft. Other than a clear preference for free chain termini in the case of exopeptidases, the substrate-binding sites exhibit no strict specificities. Instead, their subsite preferences arise more from specific exclusions of substrate type. This presents a challenge for the design of inhibitors to target a specific cathepsin: only the cumulative effect of an assembly of inhibitor fragments can produce the desired result. The small number of papain-like lysosomal cysteine proteases (11 human enzymes are known) and the small number of substrate-binding sites calls for a innovative and empirical approach. PMID- 12554932 TI - Ping-pong cross-validation in real space: a method for increasing the phasing power of a partial model without risk of model bias. AB - Experimental phases could only be obtained to 4.4 A resolution for crystals of the SecA translocation ATPase. Density modification of these phases exploiting the 65% solvent content of the crystal produced a map from which an approximate backbone model could be built for 80% of the structure. Combining the phases inferred from this partial model with the MIR phases and repeating the density modification produced an improved map from which a more complete backbone model could be built. However, this procedure converged before yielding a map, that allowed unambiguous sequence assignment for the majority of the protein molecule. In order to avoid the likely model bias associated with a speculative attempt at sequence assignment, a real-space cross-validation procedure was employed to facilitate completion of the crystal structure based on partial model phasing. The protein was partitioned into two disjoint sets of residues. Models in which the side chains were built for residues in one of the two sets were used for phase combination and density modification in order to produce improved electron density for interpretation of residues in the other set that had not been included in the model. Residues in the two sets were therefore omitted from the model in alternation except at sites where the side chain could be identified definitively based on phasing with the other set. This ping-pong cross-validation procedure allowed partial model phasing to be used to complete the crystal structure of SecA without being impeded by model bias. These results show that the structure of a large protein molecule can be solved with exclusively low resolution experimental phase information based on intensive use of partial model phasing and density modification. Real-space cross-validation can be applied to reduce the risk of model bias associated with partial model phasing, streamlining this approach and expanding its range of applicability. PMID- 12554933 TI - Structural characterization of an MJ1267 ATP-binding cassette crystal with a complex pattern of twinning caused by promiscuous fiber packing. AB - ATP-binding cassettes represent the motor domains in ABC transporters, a superfamily of integral membrane-protein pumps that couple the hydrolysis of ATP to transmembrane solute translocation. A crystal of a Mg-ADP complex of the MJ1267 ATP-binding cassette was obtained that produced a diffraction pattern characterized by pathological streaking of the spots in the a* x b* plane. While the Laue symmetry of the diffraction pattern was P3;1m, the crystal was determined to be twinned based on intensity statistics, molecular-replacement analysis and difference Fourier analysis of an engineered single-site methylmercury derivative. The unit cell contains three similar 3(1) fibers, with two of them related by primarily translational non-crystallographic symmetry (NCS) and the third related to the first two by approximate twofold screw operations whose rotational components are very similar to the twinning operator. The promiscuous packing of these 3(1) fibers, which make both parallel and antiparallel interactions in the primary crystal lattice, can explain the twinning tendency based on the ability of the twin-related lattices to interact with one another while making only one slightly sub-optimal intermolecular contact per unit cell in the boundary region. The promiscuous fiber packing can also explain the streaking in the diffraction pattern based on the ability to form a variety of different lattices with similar inter-fiber packing interactions. The crystal structure was refined as a twin in space group P3(1) using the program CNS, yielding a free R factor of 28.9% at 2.6 A and a refined twin fraction of 0.50. The structure shows a rigid-body rotation of the ABC transporter-specific alpha-helical subdomain (ABCalpha subdomain) in MJ1267 compared with the conformation observed for the same protein in a C2 crystal lattice; this observation suggests that the ABCalpha subdomain is flexibly attached to the F1-type ATP-binding core of the ATP-binding cassette when Mg-ADP is bound at the active site. PMID- 12554934 TI - Membrane-protein crystallization in cubo: temperature-dependent phase behaviour of monoolein-detergent mixtures. AB - The lipidic cubic phase of monoolein has proved to be a matrix well suited to the production of three-dimensional crystals of membrane proteins. It consists of a single continuous bilayer, which is contorted in three-dimensional space and separates two distinct water channels. It has previously been proposed that on the addition of precipitants, membrane proteins embedded in the cubic phase migrate through the matrix to nucleation sites and that this process is dependent upon the stability of the lipidic cubic phase. Here, the effect of detergent type (C(8)-C(12) glucosides, C(8)-C(12) maltosides and C(7) thioglucoside) and concentration (1-3x the critical micelle concentration; CMC) on cubic phase stability are reported in the form of the temperature-dependent phase behaviour (268-313 K) in 40% aqueous solution. The results are tabulated to show the best monoolein (MO)-detergent mixtures, mixing temperatures and crystallization temperatures identified. Monoolein-detergent mixtures suited for low-temperature in cubo crystallization of temperature-sensitive proteins are also reported for the first time. These mixtures can be prepared at low temperatures (mixed at 0.6). The system may be understood as an OD (order-disorder) structure based on a layer with layer group P(c)c2 and cell a(o) approximately 4.7, b approximately 26.1 and c approximately 14.4 A. At 338 K the layer stacking is random, but with decreasing temperature the build-up of an orthorhombic MDO (maximal degree of order) structure with cell a(1) = 2a(o), b(1) = b, c(1) = c and space group Pcc2 is begun (at approximately 301 K). The superposition structure of the OD system at T = 286 (1) K with space group Bmmb and cell a = 2a(o), b = b and c = c/2 owes its cohesion to van der Waals interactions between the AA chains and to three types of hydrogen bonds of varied strength between U-U and U-AA. Before reaching completion, this MDO structure is transformed, at 282 K, into a monoclinic one with cell a(m) = -a(o) + c/4, b(m) = b, c(m) = -2(a(o) + c/2), space group P2(1)/c, spontaneous deformation approximately 2 degrees, and ferroelastic domains. This transformation is achieved in two steps: first a furtive triggering transition, which is not yet fully understood, and second an improper ferroelastic transition. At approximately 233 K, the system reaches its ground state (cell a(M) = a(m), b(M) = b, c(M) = c(m) and space group P2(1)/c) via an irreversible transition. The phase transitions below 338 K are described by a model based on the interaction of two thermally activated slip systems. The OD structure is described in terms of a three-dimensional Monte Carlo model that involves first- and second neighbour interactions along the a axis and first-neighbour interactions along the b and c axes. This model includes random shifts of the chains along their axes and satisfactorily accounts for most features that are seen in the observed diffraction pattern. PMID- 12554975 TI - Phosphonoacetic acid as a building block in supramolecular chemistry: salts with organic polyamines. AB - Phosphonoacetic acid, (HO)(2)P(O)CH(2)COOH, forms adducts with a range of amines. The acid component in these adducts may be the neutral molecule C(2)H(5)O(5)P, the mono-anion (C(2)H(4)O(5)P)(-) or the di-anion (C(2)H(3)O(5)P)(2-). The substructure formed by the acid component takes the form of simple chains in compounds (1)-(3), which are the 1:1 adducts formed with 1,4 diazabicyclo[2.2.2]octane, 4,4'-bipyridyl and 1,3-trimethylenedipiperidine, respectively. These adducts contain C(2)H(5)O(5)P, (C(2)H(4)O(5)P)(-) and (C(2)H(3)O(5)P)(2-), respectively, although (3) is solvated by a mixture of methanol and water. The (C(2)H(4)O(5)P)(-) anion substructure in (4), which is the adduct formed with meso-5,5,7,12,12,14-hexa-C-methyl-1,4,8,11 tetraazacyclotetradecane, is a chain of spiro-fused rings, while the substructure in (5), which is the adduct formed with 2,2'-dipyridylamine, is a chain of edge fused rings. In (6), the adduct formed with 1,2-bis(4'-pyridyl)ethane, the anion substructure is two-dimensional. The chain substructures are linked by the amine units into two-dimensional structures in (1) and (4) and into three-dimensional frameworks in (2), (3) and (5), while the anion sheets in (6) are likewise linked by the amine units into a three-dimensional framework. PMID- 12554976 TI - Salts of maleic and fumaric acids with organic polyamines: comparison of isomeric acids as building blocks in supramolecular chemistry. AB - Maleic acid and fumaric acid both readily form adducts with organic diamines: maleic acid usually forms 2:1 adducts with bases, while fumaric acid usually forms 1:1 adducts, and the supramolecular structures within the two series are not simply related. The 1:2 adducts formed by 1,2-bis(4-pyridyl)ethane and by 4,4'-bipyridyl, respectively, with maleic acid, compounds (1) and (2), are salts [(diamine)H(2)(2+)].[(C(4)H(3)O(4))(-)](2) in which the cations lie across a centre of inversion and a twofold rotation axis, respectively. The ions are linked by N-H...O hydrogen bonds into three-ion aggregates, which are further linked by C-H...O hydrogen bonds into two- and three-dimensional arrays, respectively. In the fumarate salts formed by 2,2'-dipyridylamine (1:1) and 1,4 diazabicyclo[2.2.2]octane (1:2), compounds (3) and (4), the ionic components are linked into molecular ladders. The 1:1 adduct of 4,4'-bipyridyl and fumaric acid, compound (5), contains two neutral components, both of which lie across centres of inversion; these components are linked into chains by a single O-H...N hydrogen bond and thence into sheets by C-H...O hydrogen bonds. The corresponding adduct formed by 1,4-diazabicyclo[2.2.2]octane, compound (6), is a salt that again contains chains linked into sheets by C-H...O hydrogen bonds. In the 1:1 adducts, compounds (7), (8) and (10), that are formed between 1,2-bis(4 pyridyl)ethane, 4,4'-trimethylenedipyridine and hexamethylenetetramine, respectively, with fumaric acid, and in the 1:2 adduct, compound (9), of 2,2' dipyridylamine and maleic acid, the chains that are generated by the hard hydrogen bonds are linked by C-H...O hydrogen bonds to form, in each case, a single three-dimensional framework. In the 1:1 adduct, compound (11), of 2,2' bipyridyl and fumaric acid the hydrogen bonds generate two interwoven three dimensional frameworks. PMID- 12554977 TI - (1R,3S)-Camphoric acid as a building block in supramolecular chemistry: adducts with organic polyamines. AB - (1R,3S)-Camphoric acid [(1R,3S)-1,2,2,-trimethylcyclopentane-1,3-dicarboxylic acid, C(10)H(16)O(4)] forms adducts with a range of amines in which the acid component may be the neutral molecule, the mono-anion (C(10)H(15)O(4))(-) or the di-anion (C(10)H(14)O(4))(2-). The structures generated by the hard hydrogen bonds take the form of chains in the 1:1 adducts (II) and (III) formed with 4,4' bipyridyl and 1,2-bis(4-pyridyl)ethane. There are single sheets in the hydrated 1:1 adduct (IV) formed with 1,4-diazabicyclo[2.2.2]octane, and pairwise interwoven sheets in the 2:1 adduct (V) formed with hexamethylenetetramine. Three dimensional frameworks are present in the salt-like 1:1 adduct (VI) formed with piperazine and in the hydrated 3:1 adduct (VII) formed with N,N' dimethylpiperazine. This latter adduct contains both neutral C(10)H(16)O(4) and anionic (C(10)H(15)O(4))(-) units. In (II), (III) and (IV), the chain and sheet substructures are linked by C-H...O hydrogen bonds to form three-dimensional frameworks. The monoclinic polymorph of camphoric acid itself (I) has been reinvestigated. PMID- 12554978 TI - Hydrogen bonding in enantiomeric versus racemic mono-carboxylic acids; a case study of 2-phenoxypropionic acid. AB - The structural and thermodynamic backgrounds for the crystallization behaviour of racemates have been investigated using 2-phenoxypropionic acid (PPA) as an example. The racemate of PPA behaves normally and forms a racemic compound that has a higher melting point and is denser than the enantiomer. Low-temperature crystal structures of the pure enantiomer, the enantiomer cocrystallized with n alkanes and the racemic acid showed that hydrogen-bonded dimers that form over crystallographic symmetry elements exist in all but the structure of the pure enantiomer. A database search for optically pure chiral mono-carboxylic acids revealed that the hydrogen-bonded cyclic dimer is the most prevalent hydrogen bond motif in chiral mono-carboxylic acids. The conformation of PPA depends on the hydrogen-bond motif; the antiplanar conformation relative to the ether group is associated with a catemer hydrogen-bonding motif, whereas the more abundant synplanar conformation is found in crystals that contain cyclic dimers. Other intermolecular interactions that involve the substituent of the carboxylic group were identified in the crystals that contain the cyclic dimer. This result shows how important the nature of the substituent is for the crystal packing. The differences in crystal packing have been related to differences in melting enthalpy and entropy between the racemic and enantiomeric acids. In a comparison with the equivalent 2-(4-chlorophenoxy)-propionic acids, the differences between the crystal structures of the chloro and the unsubstituted acid have been identified and related to thermodynamic data. PMID- 12554979 TI - An X-ray diffraction study of partially ordered electron density in clathrates of Dianin's compound that include simple carboxylic acids. AB - The nature of the inclusions in ten clathrate complexes of Dianin's compound have been investigated by the use of electron-density difference maps. The guest species are the first eight straight-chain carboxylic acids, formic to octanoic, a branched-chain acid (dimethylacetic acid) and trifluoroacetic acid. The point group symmetry of the clathrate cavity, 3, is satisfied by partial occupation of symmetry-related sites by two included molecules in the case of formic, acetic and trifluoroacetic acids and by a single molecule in the remainder. Hydrogen bonding requirements in the case of formic and acetic acids are satisfied by dimer formation; in the trifluoroacetic acid complex the two acid molecules form hydrogen bonds to framework O atoms at either end of the cavity. The adoption of gauche conformations in heptanoic and octanoic acid chains shortens them sufficiently that they fit the cavity with only slight distortion. PMID- 12554980 TI - Single-crystal-to-single-crystal photodimerization of 4-chlorocinnamoyl-O,O' dimethyldopamine. AB - [2+2]Photodimerization of the title compound, C(19)H(20)ClNO(3), has been observed in situ by single-crystal X-ray diffraction. Pairs of monomers related by centers of symmetry have parallel C=C bonds at a C.C distance of 4.155 (17) A. Irradiation of a single crystal with a mercury lamp achieves 100% conversion to the dimer. Redetermination of the structure during the course of the reaction revealed a linear correlation between the percent conversion to the dimer and the decrease in the cell Volume. The displacement parameters for the pure dimer structure are substantially smaller than those for the pure monomer structure. The dimerization reaction is also induced by irradiation with X-rays, the induction being stronger with Cu Kalpha than with Mo Kalpha radiation. PMID- 12554981 TI - Twinning in natural melilite simulating a fivefold superstructure. AB - Additional reflections seemingly leading to a tetragonal fivefold supercell have been observed in a number of melilites. A careful examination of the collected intensity data reveals that this feature is due to the coexistence of two twin domains related by a (120) twin plane. Reflections of the first domain (I) with h(b)(2) + k(b)(2) = 5n overlap those of the second domain (II) when both the following additional conditions are verified: -h(b) + 2k(b) = 5n and 2h(b) + k(b) = 5n. Indices of the superimposed reflections are h(b,I) k(b,I) l(b,I) for the first domain, and h(b,II) = 3/5h(b,I) + 4/5k(b,I), k(b,II) = 4/5h(b,I) - 3/5k(b,I), l(b,II) = l(b,I) for the second domain. The non-crystallographic pseudosymmetry in the melilite structure is discussed. PMID- 12554985 TI - The winds of change. PMID- 12554986 TI - Intramural pregnancy. PMID- 12554987 TI - Bowel injury in gynecologic laparoscopy. AB - STUDY OBJECTIVE: To review surveys of the last decade on bowel injuries to evaluate the prevalence, causes, management, and outcomes of these events occurring during or as a result of laparoscopy. DESIGN: Retrospective evaluation (Canadian Task Force classification II-2). SETTING: Surveys and databases. PATIENTS: None. INTERVENTION: Data analysis. MEASUREMENTS AND MAIN RESULTS: Combined data show that diagnostic and minor operative laparoscopy are associated with a 0.08% risk of bowel injury, and in major operative laparoscopy the risk increases to 0.33%. Injuries occurring during access and operative procedure decrease significantly with experience, but even in experienced hands injury during access cannot be avoided. Delayed diagnosis remains a major problem. Up to 15% of these injuries are not diagnosed during laparoscopy, and one of five cases of delayed diagnosis results in death. Perioperative diagnosis and immediate repair by laparoscopy or laparotomy reduce the likelihood of severe complications and consequently medicolegal actions. CONCLUSIONS: Several surveys on complications of gynecologic laparoscopy tend to underestimate the risk of bowel injury. Prevention starts by awareness that such injury is an inherent risk of the technique, even in hands of experienced surgeons. PMID- 12554988 TI - The minilaparoscope as a tool for localization and preparation for cannula insertion in patients with multiple previous abdominal incisions or umbilical hernia. AB - STUDY OBJECTIVE: To access a method that attempts to reduce the risk of bowel and blood vessel trauma in entry-risk patients during laparoscopy. DESIGN: Three-year observational study (Canadian Task Force classification II-2). SETTING: Tertiary university hospital. PATIENTS: Thirty-one women defined as entry-risk due to previous multiple abdominal surgeries or repair of umbilical hernia. INTERVENTION: Laparoscopy performed with a 2 mm minilaparoscope inserted at Palmer's point, a midclavicular position under the lower left rib. MEASUREMENTS AND MAIN RESULTS: In most cases the minilaparoscope was used to inspect the anterior abdominal wall for adhesions. When a location free from adhesions was seen or created by adhesiolysis, the 5- or 10-mm cannula was inserted, followed by the laparoscope. Periumbilical adhesions were more common after previous longitudinal incisions. There were no complications. CONCLUSIONS: Minilaparoscopy is safe and effective for identifying and preparing a proper cannula insertion point. This may be useful for avoiding bowel or other cannula-related trauma in women at high risk for such a complication. PMID- 12554989 TI - A randomized study comparing endometrial cryoablation and rollerball electroablation for treatment of dysfunctional uterine bleeding. AB - STUDY OBJECTIVE: To determine the effectiveness of endometrial cryoablation in comparison with rollerball electroablation. DESIGN: Prospective, randomized study (Canadian Task Force classification I). SETTING: Ten university and private medical centers in the United States. PATIENTS: Two hundred seventy-nine women with menorrhagia due to benign causes. INTERVENTION: Endometrial ablation using a Her Option cryoablation device in 193 women and rollerball electroablation in 86. MEASUREMENTS AND MAIN RESULTS: Women treated by cryoablation received significantly less general anesthesia (46%) than those treated by electroablation (92%). Subjects maintained menstrual diaries for at least one cycle before and for 12 months after the procedure. Success was defined as reduction of menstrual bleeding to a score of 75 or less in the absence of retreatment. Success rates in the cryoablation and electroablation groups were 77.3% and 83.8%, respectively. Bleeding declined by 92% and 94%, respectively. Both procedures led to significant improvements in a broad range of symptoms including menses-related pain, mood, and overall improvement in quality of life. CONCLUSIONS: Endometrial cryoablation is a safe and effective procedure in treatment of dysfunctional uterine bleeding. Its advantages include technical ease of performance, direct ultrasonographic view of depth of ablation, little anesthetic, and avoidance of potential complications related to distention media. PMID- 12554990 TI - Endometrial ablation with a new thermal balloon system. AB - STUDY OBJECTIVE: To assess the efficacy of Thermablate EAS, a new, simple, hand held, portable endometrial ablation instrument, in the treatment of menorrhagia. DESIGN: Retrospective observational study (Canadian Task Force classification II 1). SETTING: Urban hospital and private clinic facilities in Bombay, India. PATIENTS: Sixteen women with menorrhagia. INTERVENTION: Endometrial ablation with the Thermablate EAS. MEASUREMENTS AND MAIN RESULTS: Follow-up at 6 months showed eight patients (50%) to have amenorrhea and six (38%) hypomenorrhea. The only failure was in a patient with cystic hyperplasia. No complications occurred. CONCLUSIONS: Thermablate EAS is a promising instrument for endometrial ablation. PMID- 12554991 TI - Frequency and laparoscopic management of ovarian remnant syndrome. AB - STUDY OBJECTIVE: To report the frequency and outcome of laparoscopy in women with chronic pelvic pain and/or pelvic mass who were found to have ovarian remnants. DESIGN: Cohort study. (Canadian Task Force classification II-2). SETTING: University-affiliated hospital. PATIENTS: One hundred nineteen women who had had hysterectomy and oophorectomy. INTERVENTION: Laparoscopic surgery. MEASUREMENTS AND MAIN RESULTS: Ovarian remnants were known in 5 and were found intraoperatively in 21 patients (18%). These 26 women had undergone at least one laparoscopy in an attempt to remove the remnants. After the ureter was identified, ovarian remnants were dissected and removed from the retroperitoneum laparoscopically with minimal risk of vessel or visceral injury. There were no intraoperative or postoperative complications and no conversions to laparotomy. In addition to ovarian remnants, adhesions were found in 19 women, endometriosis in 4, and no other pathology in 3. Twenty women had complete relief of symptoms. At follow-up of 1 to 8 years (mean 5 yrs), six underwent repeat laparoscopy for persistent pain; one had recurrent ovarian remnant. CONCLUSIONS: Ovarian remnant syndrome is not an infrequent complication after hysterectomy and oophorectomy in women with endometriosis. PMID- 12554992 TI - Two-year experience with laparoscopic pelvic floor repair. AB - STUDY OBJECTIVE: To evaluate the cumulative experience at our institution of laparoscopic pelvic floor repair to treat genital prolapse and associated symptoms. DESIGN: Retrospective analysis (Canadian Task Force classification II 2). SETTING: University hospital. PATIENTS: Seventy-three consecutive women treated surgically for symptomatic genital prolapse. INTERVENTION: Surgical treatment was site specific depending on findings on physical examination. Anterior compartment defects were treated by laparoscopic paravaginal repair, laparoscopic Burch colposuspension, or transvaginal anterior vaginal repair. Defects in the posterior compartment were treated by a combination of laparoscopic supralevator repair, laparoscopic vaginal vault suspension, enterocele sac invagination or excision, and transvaginal posterior vaginal repair. Anatomic defects in the apical compartment were primarily treated by laparoscopic vaginal vault suspension and enterocele sac excision. Patients whose anatomic anomalies contained elements of anterior, posterior, and apical compartments were classified in a global group. MEASUREMENTS AND MAIN RESULTS: Preoperatively, prolapse was considered as an attachment or fascial defect at DeLancey level I, II, or III. Each was then quantified by the pelvic organ prolapse quantification (POPQ) system and compartmentalized according to site of the major defect. Women were assessed by physical examination and repeat POPQ staging 6 weeks postoperatively and every 6 months thereafter. A standard interview was administered to assess functional status. Major complications occurred in 4.1% of women. Objective and subjective cure rates were 90% at 2 years. CONCLUSIONS: Laparoscopic pelvic floor repair is an effective procedure with low morbidity. It should play a primary role in surgical management of DeLancey levels I and II attachment defects. For fascial defects, in particular DeLancey level II anteriorly and posteriorly, it should be complemented with vaginal repair. PMID- 12554993 TI - High frequency of endometrial polyps in endometriosis. AB - STUDY OBJECTIVE: To evaluate the effectiveness of hysteroscopy and the frequency of endometrial polyps in women with endometriosis. DESIGN: Retrospective clinical study (Canadian Task Force classification II-2). SETTING: Infertility unit at a university-affiliated hospital. PATIENTS: One hundred eighty-three infertile women, 92 with endometriosis and 91 controls without the disease. INTERVENTION: Laparoscopy and scoring of endometriosis according to the American Fertility Society classification, and confirmation of endometrial polyps by pathologic examination. MEASUREMENTS AND MAIN RESULTS: There was no significant difference between groups with regard to age, mean duration of infertility, and frequency of primary or secondary infertility. Endometrial polyps were found in 43 women (46.7%) with endometriosis and in 15 controls (16.5%, p = 0.0000). Their frequency did not differ significantly according to stage of endometriosis. CONCLUSIONS: We strongly recommend hysteroscopy if endometriosis is detected in a woman undergoing evaluation for infertility, even if hysterosalpingography and transvaginal ultrasonography do not suggest endometrial polyps. PMID- 12554994 TI - Short- and long-term clinical results of laparoscopic-assisted vaginal hysterectomy and total abdominal hysterectomy. AB - STUDY OBJECTIVE: To compare short- and long-term clinical results of laparoscopic assisted vaginal hysterectomy (LAVH) and total abdominal hysterectomy (TAH). DESIGN: Retrospective cohort study (Canadian Task Force classification II-1). SETTING: University-affiliated hospital. PATIENTS: One hundred fifty women who underwent LAVH and 146 who underwent TAH. INTERVENTION: Hysterectomy. MEASUREMENTS AND MAIN RESULTS: Blood loss during surgery, narcotic analgesic consumption, duration of hospital stay, and convalescence time were significantly higher for women who underwent TAH than for those who underwent LAVH (p <0.05). Operating time was significantly longer for LAVH than for TAH (152.2 +/- 32.4 vs 96.5 +/- 29.6 min, p = 0.014). Eight-year follow-up showed no statistically significant differences in vaginal vault prolapse, cystocele, rectocele, enterocele, postcoital spotting, and cuff granulation between procedures (p >0.05). CONCLUSIONS: Although short-term clinical results revealed some statistically significant differences between LAVH and TAH, long-term follow-up recorded similar frequencies of surgical sequelae. PMID- 12554995 TI - Laparoscopy for adnexal masses in the second trimester of pregnancy. AB - STUDY OBJECTIVE: To assess the safety of laparoscopic treatment of adnexal masses in the second trimester of pregnancy. DESIGN: Retrospective chart review (Canadian Task Force classification II-3. SETTING: Large tertiary care medical center. PATIENTS: Eleven women. INTERVENTION: Laparoscopic surgery. MEASUREMENTS AND MAIN RESULTS: All masses were identified by ultrasound. The average gestational age at the time of surgery was 17 weeks, 4 days. In seven women the primary cannula was inserted in the left upper quadrant of the abdomen. Procedures were eight ovarian cystectomies, two oophorectomies, and one exploratory laparoscopy. Average operating time was 135 minutes (range 95-195 min). Average time exposed to carbon dioxide was 78 minutes (range 59-135 min). None of the masses was malignant. There were no fetal complications or malformations. No patients had preterm labor and all delivered at term. CONCLUSIONS: The increasing number of reported cases and our experience suggest that laparoscopic treatment of adnexal masses in the second trimester is safe and effective, but the surgeon must be skilled in advanced techniques of laparoscopic surgery. PMID- 12554996 TI - Alteration of sexual function after classic intrafascial supracervical hysterectomy and total hysterectomy. AB - STUDY OBJECTIVE: To investigate possible advantages of classic intrafascial supracervical hysterectomy (CISH) over total hysterectomy with respect to sexual function. DESIGN: Retrospective study (Canadian Task Force classification II-2). SETTING: Department of obstetrics and gynecology at a university teaching hospital. PATIENTS: One hundred eight women who had undergone CISH and 125 who had total hysterectomy. INTERVENTION: A standard questionnaire sent to all 233 patients a minimum of 12 and maximum of 24 months after hysterectomy. MEASUREMENTS AND MAIN RESULTS: Data obtained on sexual function were classified by changes (improved, no change, deteriorated) before and after hysterectomy. No significant difference was found between groups in mean time interval between surgery and first intercourse. Overall, over two-thirds of women experienced either no change or improvement in sexual function regardless of which procedure was performed, with no statistically significant differences in changes in libido, frequency of coitus, frequency of orgasm, and degree of orgasm. CONCLUSIONS: Women who underwent CISH and total hysterectomy had no statistically significant differences with regard to postoperative sexual function. PMID- 12554997 TI - Comparison of peritoneal oxidative stress during laparoscopy and laparotomy. AB - STUDY OBJECTIVE: To identify oxidative stress in peritoneum during laparoscopic and open surgery by measuring products of lipid peroxidation, and to determine whether surgical approach influences the type of oxidative metabolite synthesized. DESIGN: Retrospective analysis (Canadian Task Force classification II-2). SETTING: University-affiliated hospital. PATIENTS: Twenty-eight consecutive women with uterine myomas or ovarian cysts. INTERVENTION: Laparoscopic or open surgery (14 patients each). MEASUREMENTS AND MAIN RESULTS: We obtained 1 x 1-cm squares of peritoneum at the beginning and end of surgical procedures away from sites of surgery. 8-Isoprostaglandin F(2alpha), hydroxyeicosatetranoic acids (HETEs), and malondyaldehyde (MDA) were measured by enzyme-immunoassay, high-performance liquid chromatography, and thiobarbituric acid adduction method, respectively. Comparisons showed significant increases in 5-HETE and 8-prostane in the laparoscopy group, which were correlated with duration of pneumoperitoneum and volume of carbon dioxide (CO(2)) insufflated, respectively. In the laparotomy group only MDA rose significantly related to duration of surgery. CONCLUSIONS: Lipid peroxidation was observed in peripheral peritoneum during laparoscopic surgery, mediated through noncyclooxygenase and lipoxygenase pathways, and appears to be due to effects of CO(2) pneumoperitoneum. Biochemical reactions were also observed in the laparotomy group, but are thought to be related to mechanisms other than lipid peroxidation. PMID- 12554998 TI - Comparison of classic and endoscopic lymphadenectomy for staging breast cancer. AB - STUDY OBJECTIVE: To compare endoscopic and classic axillary lymphadenectomy staging of breast cancer with respect to operation-induced changes such as seroma formation, pain, neurologic sensations, lymphedema, infection, and reduction of shoulder-arm mobility. DESIGN: Prospective, randomized study (Canadian Task Force classification I). SETTING: University-affiliated hospital. PATIENTS: Eighty consecutive women with histopathologically confirmed invasive breast cancer who had clinically and sonographically negative axillary lymph nodes (<1 cm). INTERVENTION: Classic and endoscopic lymphadenectomies. MEASUREMENTS AND MAIN RESULTS: We attempted to obtain 10 axillary lymph nodes/patient. After 1, 3, 5, 7, 9, 42, and 84 days, clinical and ultrasonographic examinations were conducted to evaluate operation-induced changes. Short-term results showed that, with endoscopic technique, a representative number of axillary lymph nodes was removed, with reduced axillary infiltration and seroma induction, as well as less impaired shoulder-arm mobility. During the first month, postoperative infection, lymphedema, and neurologic complaints were comparable in both groups, with more stretching pain in the classic group and predominantly paresthesia in the endoscopic group. After 3 months no differences in postoperative complications were detected. CONCLUSIONS: Endoscopic axillary lymphadenectomy avoids short-term reduction of shoulder-arm mobility. Long-term studies are necessary to prove if this technique is as safe as the classic procedure with regard to local axillary recurrence. If so, endoscopy could become the method of choice for staging breast cancer in women with clinically negative lymph nodes. PMID- 12554999 TI - Preliminary results of laparoscopic modified radical hysterectomy in early invasive cervical cancer. AB - STUDY OBJECTIVE: To compare laparoscopic modified radical hysterectomy (LMRH) with traditional modified radical hysterectomy (MRH) in women with early invasive cervical cancer. DESIGN: Prospective, nonrandomized study (Canadian Task Force classification II-1). SETTING: Chi Mei Foundation Medical Center, Tainan, Taiwan. PATIENTS: Ten women with cervical cancer stage Ia2 to Ib1 (50%) and in close proximity to the FRA9E CFS. FISH analysis determined that PAPPA was contained within the distal end of FRA9E. Characterization of FRA9E determined that aphidicolin induced instability extended over 9 Mb, identifying FRA9E as the largest CFS characterized to date. Comprehensive LOH analysis revealed several distinct peaks of LOH within FRA9E. Semiquantitative RT-PCR analysis of 16 genes contained within FRA9E indicated that genes showing LOE in ovarian tumors coincided with regions of high LOH. PAPPA displayed the most significant loss (72%). This study provides evidence to suggest that instability within FRA9E may play an important role in the development of ovarian cancer and lends further support for the hypothesis that CFSs may be causally related to cancer. PMID- 12555073 TI - An inhibitory function for JNK in the regulation of IGF-I signaling in breast cancer. AB - Insulin-like growth factor-I receptor (IGF-IR) is frequently overexpressed in a variety of cancer types. Since many breast tumors and cancer cell lines overexpress IGF-IR, we tested IGF-I effects on chemotherapy-treated breast cancer cells. IGF-I protects from chemotherapy-induced apoptosis, suggesting that overlapping signaling pathways modulate IGF-I and chemotherapy treatment outcomes. Taxol and other chemotherapy drugs induce c-Jun N-terminal kinase (JNK), a kinase that conveys cellular stress and death signals. Notably, in this paper we show that IGF-I alone induces a potent JNK response and this activity is reversed by inhibition of phosphatidylinositol 3-kinase (PI 3-kinase) with LY294002 in MCF-7 but not T47D cells. Cotreatment of cells with chemotherapy and IGF-I leads to additive JNK responses. Using cells overexpressing Akt, we confirm that IGF-I-mediated survival is Akt dependent. In contrast, overexpression of JNK significantly enhances Taxol-induced apoptosis and inhibits IGF-I survival effects. Further, JNK attenuates anchorage-independent growth of MCF-7 cells. The inhibitory effect of JNK appears to be mediated by serine phosphorylation of IRS 1 (insulin receptor substrate) since both Taxol and IGF-I treatment enhanced Ser(312) IRS-1 phosphorylation, while LY294002 blocked IGF-I-mediated phosphorylation. Taken together, these data provide a mechanism whereby stress or growth factors activate JNK to reduce proliferation and/or survival in breast cancer cells. PMID- 12555075 TI - Establishment of latrunculin-A resistance in HeLa cells by expression of R183A D184A mutant beta-actin. AB - Actin plays central roles in cell motility through formation of the actin cytoskeleton. Recently, the very intriguing possibility that actin also contributes to processes in the cell nucleus has been emerging. To dissect its dynamics and functions, several actin-disrupting drugs have been widely and effectively employed. Among them, latrunculin-A has proved particularly useful, supplanting the classical drug cytochalasin-D. One reason is that latrunculin-A appears to bind only to actin monomers impairing the nucleotide exchange, the mode being simpler than with cytochalasin. This property may be especially crucial when studying actin functions as a monomer, as suggested for nuclear actin. Very importantly, actin mutations that cause cells to become resistant to the effects of latrunculin-A have been identified in budding yeast. However, it remains controversial as to whether all of the various phenotypes observed with latrunculin in mammalian cells more complicated than yeast are truly the consequence of its specific actions against actin. Here, we show that the expression of R183A D184A mutant beta-actin specifically confers resistance to the effects of latrunculin-A on actin cytoskeleton formation and cell growth in HeLa cells. The established system provides a strong tool to address the various functions of actin in mammalian cells. PMID- 12555074 TI - CACNA2D2-mediated apoptosis in NSCLC cells is associated with alterations of the intracellular calcium signaling and disruption of mitochondria membrane integrity. AB - The CACNA2D2 gene, a new subunit of the Ca(2+)-channel complex, was identified in the homozygous deletion region of chromosome 3p21.3 in human lung and breast cancers. Expression deficiency of the CACNA2D2 in cancer cells suggests a possible link of it to Ca(2+) signaling in the pathogenesis of lung cancer and other cancers. We investigated the effects of overexpression of CACNA2D2 on intracellular Ca(2+) contents, mitochondria homeostasis, cell proliferation, and apoptosis by adenoviral vector-mediated wild-type CACNA2D2 gene transfer in 3p21.3-deficient nonsmall cell lung cancer cell lines. Exogenous expression of CACNA2D2 significantly inhibited tumor cell growth compared with the controls. Overexpression of CACNA2D2 induced apoptosis in H1299 (12.5%), H358 (13.7%), H460 (22.3%), and A549 (50.1%) cell lines. Levels of intracellular free Ca(2+) were elevated in AdCACNA2D2-transduced cells compared with the controls. Mitochondria membrane depolarization was observed prior to apoptosis in Ad-CACNA2D2 and Adp53 transduced H460 and A549 cells. Release of cyt c into the cytosol, caspase 3 activation, and PARP cleavage were also detected in these cells. Together, these results suggest that one of the pathways in CACNA2D2-induced apoptosis is mediated through disruption of mitochondria membrane integrity, the release of cyt c, and the activation of caspases, a process that is associated with regulation of cytosolic free Ca(2+) contents. PMID- 12555076 TI - AXIN1 mutations but not deletions in cerebellar medulloblastomas. AB - Medulloblastoma is a malignant, invasive embryonal tumour of the cerebellum which manifests preferentially in children. A subset of cases is associated with colon cancer and APC germline mutations (Turcot syndrome), and APC and beta-catenin point mutations occur in up to 10% of sporadic cases, indicating the involvement of the Wnt pathway in the development of medulloblastoma. In 39 sporadic cerebellar medulloblastomas screeened for alterations in the AXIN1 gene, another component of the Wnt pathway, we found missense AXIN1 mutations in two tumours, CCC-->TCC at codon 255 (exon 1, Pro-->Ser) and TCT-->TGT at codon 263 (exon 1, Ser-->Cys). Furthermore, the A allele at the G/A polymorphism at nucleotide 16 in intron 4 was significantly over-represented in medulloblastomas (39 cases; G 0.76 vs-A 0.24) compared to healthy individuals (86 cases; G 0.91 vs A 0.09; P=0.0027). RT-PCR revealed large deletions in the AXIN1 gene in 5/12 (42%) medulloblastomas, consistent with a previous report. However, we observed such deletions at a similar frequency also in normal brain tissue (6/12, 50%). Since there are multiple complementary, inverted sequences present in the AXIN1 gene, these large deletions may represent RT-PCR errors due to stem-loop secondary structures. PMID- 12555078 TI - Watching flipping junctions. PMID- 12555079 TI - Snapshots of transition states? PMID- 12555080 TI - Shape can be seductive. PMID- 12555081 TI - Second messenger control of chromatin remodeling. PMID- 12555082 TI - Tick-tock goes the acetylation clock. PMID- 12555083 TI - A folding machine for many but a master of none. PMID- 12555084 TI - Of men and machines. PMID- 12555088 TI - The specificity of the clonal selection theory. PMID- 12555089 TI - Thymic output: a bad TREC record. PMID- 12555090 TI - To T or not to T: reassessing the common lymphoid progenitor. PMID- 12555091 TI - Everything is E(Z): linking histone methylation to B cell development. PMID- 12555092 TI - Location, location, location. PMID- 12555093 TI - Toll signaling: the TIReless quest for specificity. PMID- 12555094 TI - Granzyme A: the road less traveled. PMID- 12555097 TI - Brain myths. PMID- 12555096 TI - Adaptors as central mediators of signal transduction in immune cells. AB - Adaptors are molecular scaffolds that recruit effectors, which are critical for immune cell activation. Recent work has underscored the requirement for adaptors in propagating stimulatory signals as well as their ability to inhibit immune cell function. The mechanisms by which adaptors function rely not only on the intermolecular interactions they mediate, but also on where they are localized within the cell. The use of sophisticated genetic, biochemical, cellular and imaging approaches has provided important new insights into the biology of adaptor protein function. Here we focus on T lymphocytes as a model to illustrate the critical roles adaptors play as regulators of cellular activation. PMID- 12555098 TI - Actin in a supporting role. PMID- 12555099 TI - Ephrins keep dendritic spines in shape. PMID- 12555100 TI - Insulating axons via NF-kappaB. PMID- 12555101 TI - Nosing in on the emotional brain. PMID- 12555102 TI - The master clock becomes a servant. PMID- 12555104 TI - A framework for consciousness. AB - Here we summarize our present approach to the problem of consciousness. After an introduction outlining our general strategy, we describe what is meant by the term 'framework' and set it out under ten headings. This framework offers a coherent scheme for explaining the neural correlates of (visual) consciousness in terms of competing cellular assemblies. Most of the ideas we favor have been suggested before, but their combination is original. We also outline some general experimental approaches to the problem and, finally, acknowledge some relevant aspects of the brain that have been left out of the proposed framework. PMID- 12555105 TI - Report on the development of the New South Wales Child Health Survey 2001. PMID- 12555106 TI - Burden of disease New South Wales--feasibility report. PMID- 12555107 TI - New South Wales mothers and babies 2001. PMID- 12555108 TI - Environmental carcinogens in New South Wales. PMID- 12555109 TI - Environmental carcinogen control in Australia: the need for a strategy. PMID- 12555110 TI - Research and development in carcinogen control. PMID- 12555111 TI - The identification, assessment and control of environmental carcinogens in New South Wales and Australia. PMID- 12555112 TI - Benzene: a case study of the control of a carcinogen in New South Wales. PMID- 12555113 TI - Can molecular epidemiology help us better understand the environment's role in carcinogenesis? The example of pesticides. PMID- 12555114 TI - Tobacco control in NSW: evidence supporting improved strategies to reduce exposure to environmental tobacco smoke. PMID- 12555115 TI - FactSheet: Shigellosis. PMID- 12555116 TI - Communicable diseases report, NSW: September 2002. PMID- 12555117 TI - Even bigger and better: report of the Chief Health Officer 2002. PMID- 12555118 TI - Trends in potentially avoidable mortality in NSW. PMID- 12555119 TI - The mental health outcomes and assessment tools training project: creating the foundation for improved quality of care. PMID- 12555120 TI - Monitoring child health in NSW: the New South Wales Child Health Survey 2001 and beyond. PMID- 12555121 TI - Release of the National Health Survey: summary of results, Australia 2001. PMID- 12555122 TI - FactSheet: Chlamydia. PMID- 12555123 TI - Communicable diseases report, NSW: October-December 2002. PMID- 12555127 TI - Janice M. Pfeffer Memorial Lecture. PMID- 12555128 TI - Epidemiology and screening of asymptomatic left ventricular dysfunction. AB - The traditional approach to estimating heart failure prevalence has been based on signs and symptoms of heart failure; such prevalence estimates have been in the range of 2% of persons A) were determined. Isolated protein C deficiency was found in 23% of patients from the SVT group, in 5% from the AD group, in 6.8% from the NT group, and in 1% of historical controls (P = 0.0001). The prevalence of thrombophilic genotypes and that of the other natural anticoagulants did not differ across the groups. The proportion of patients with elevated plasma HC was 66% in the AD group, 27% in the non thrombotic group, 12% in the SVT group and 4.5% in the control group (P < 0.0001). Patients with AD had higher plasma HC (24.4 +/- 23 micromol/L) than NT patients (12.3 +/- 7.7 micromol/L), SVT patients (9 +/- 4.9 micromol/L), and healthy controls (7.9 +/- 3 micromol/L) (P < 0.0001). In a logistic regression model lower protein C was independently associated with SVT, whereas elevated plasma HC was independently associated with AD. Measurement of plasma HC and protein C in MPD may identify patients more likely to suffer arterial disease and splanchnic vein thrombosis and who may require plasma HC lowering in the former case. PMID- 12555210 TI - Severe chronic neutropenia: treatment and follow-up of patients in the Severe Chronic Neutropenia International Registry. AB - Severe chronic neutropenia (SCN) is defined as an absolute neutrophil (ANC) of less than 0.5 x 10(9)/L, lasting for months or years. Congenital, cyclic, and idiopathic neutropenia are principal categories of SCN. Since 1994, the Severe Chronic Neutropenia International Registry (SCNIR) has collected data to monitor the clinical course, treatments, and disease outcomes for SCN patients. This report summarizes data for 853 patients, almost all treated with daily or alternate-day recombinant human granulocyte colony-stimulating factor (G-CSF or Filgrastim). G-CSF treatment increased the ANC overall from 0.34 x 10(9)/L +/- 0.018 pre-treatment to 3.70 x 10(9)/L +/- 0.18 during the first year of treatment. For most patients, the responses were durable with patients remaining on the same dose of G-CSF for many years. Long-term hematological observations showed stable mean leukocyte and neutrophil counts and gradually increasing hemoglobin levels. Thrombocytopenia developed in 4% of patients. As of January 1, 2000, myelodysplasia (MDS) or acute myelogenous leukemia (AML) has occurred in 35 of 387 patients with congenital neutropenia with a cumulative risk of 13% after 8 years of G-CSF treatment. This event occurred without a predictable relationship to the duration or dose of G-CSF treatment. No patients with cyclic or idiopathic neutropenia developed MDS or AML. Other important adverse events included hepatomegaly, osteoporosis, vasculitis, glomerulonephritis, and deaths in 4 of 14 cases requiring splenectomy. Growth and development and the outcome of pregnancy appeared to be unaffected by G-CSF treatment. These data indicate that congenital, cyclic, and idiopathic neutropenia can be effectively treated with long-term G-CSF. The risk of leukemia, osteoporosis, other potentially adverse events, and pregnancy outcome need to be further evaluated with continuing long term observations. PMID- 12555211 TI - Long term follow-up after splenectomy performed for immune thrombocytopenic purpura (ITP). AB - Splenectomy is the only treatment of ITP known to have "curative" effects in a substantial fraction of patients. However, the true long-term outcome is uncertain and controversial because published series have not adjusted for the duration of follow-up. This IRB-approved retrospective study included all patients with ITP who underwent splenectomy between 1988-1993 at three major medical centers and required a minimum postoperative 5-year follow-up. Complete response (CR) was defined as all postsplenectomy platelet counts >150 x 10(9)/L without treatment; partial response (PR) as platelet counts > or =50 x 10(9)/L without treatment; and failure as platelet counts <50 x 10(9)/L or receiving therapy after splenectomy. Seventy-five patients identified with ITP underwent splenectomy from 1988 to 1993. Three patients died prior to 5-year follow-up, and 56 of the 72 patients (78%) were evaluable with follow-up for five years or longer, median 7.5 years. The immediate postoperative complete remission rate was 77%; 57% of patients have remained in prolonged CR. Thirty-seven patients (66%) have not required any therapy after splenectomy. Eight patients had platelet counts >150 x 10(9)/L for 4-8.5 years before relapsing; no clear plateau was attained in the remission curve. There was no operative mortality. Ten patients (18%) reported minor postoperative bleeding episodes. No life-threatening infections, significant heart disease, or pulmonary hypertension developed after splenectomy in the 434 patient-years of follow-up. This study helps to define the long-term results of splenectomy for ITP. PMID- 12555212 TI - Access to stem cell transplantation: do women fare as well as men? AB - Women have less access to certain types of expensive treatments including renal transplantation, cardiac catheterization and diagnostic studies for lung cancer. Whether women have less access to stem cell transplantation (SCT) is not known. We evaluated allogeneic SCT data from the International Bone Marrow Transplant Registry (IBMTR) and compared them with disease incidence data from the Surveillance and Epidemiologic End Results (SEER) database. We estimated the ratio of males to females among transplanted patients with acute lymphoblastic (ALL), acute myelogenous (AML) and chronic myelogenous (CML) leukemia, diseases for which SCT is often done and compared them to male/female ratios of disease incidence. The association between gender and SCT was estimated as odds ratios (OR) with 95% confidence intervals (CI). There was no association between gender (male vs female) and the rates of SCT for individuals with AML (OR = 0.95, 95% CI = 0.89-1.02), or CML (OR = 1.0; CI = 0.90-1.1). Among patients with newly diagnosed ALL, more males underwent SCT than females (OR = 1.30, CI = 1.18-1.44). Because children with newly diagnosed ALL usually have a favorable prognosis, SCT is not generally a frontline therapy. Therefore, when we compared SCT rates to a population of children with relapsed ALL, the gender differences disappeared (OR = 1.09, CI = 0.94-1.25). We conclude that for the diagnoses where SCT is commonly used, there is no significant bias towards use in males compared to females. While boys with ALL appear to receive SCT at a higher rate, this difference is likely attributable to biological rather than social reasons. PMID- 12555213 TI - Acquired activated protein C resistance, high tissue factor expression, and hyper homocysteinemia in systemic lupus erythematosus. AB - Activated protein C resistance (APCR), high tissue factor (TF) expression, and hyper-homocysteinemia are associated with thromboembolic diseases. Thromboembolism is a frequent complication of systemic lupus erythematosus (SLE). In this study, we evaluated the prevalence of APCR, high TF, and homocysteine with correlation of the thrombotic tendency in SLE. Ninety-four SLE patients and 28 normal controls were included. APC ratio and TF antigen were measured using commercial kits. Plasma homocysteine level was measured using HPLC. The prevalence of APCR, high TF antigen level, and hyper-homocysteinemia in our SLE patients were 21.3%, 66.0%, and 23.4%, respectively. The median plasma level of TF antigen in SLE patients was 145.23 pg/mL (range, 31.00-778.50 pg/mL), which was significantly higher than the control value of 39.83 pg/mL (range, 1.55 168.50 pg/mL). The median APC ratio in SLE patients was 2.76 (range, 1.48-13.47), which was significantly lower than the control value of 3.59 (range, 0.26-5.66). The plasma level of homocysteine was not significantly different from that of control. A significant association was observed between the presence of APCR (OR = 8.59, P < 0.0001) but not with the presence of high plasma TF antigen level (OR = 1.24, P = 0.67) and thrombotic complications in SLE patients. In conclusion, APCR and high plasma TF levels are common in SLE, but a significant association was observed only between the presence of APCR and thrombosis in SLE patients. PMID- 12555214 TI - Thromboembolism preceding cancer: a correlation study. AB - Thromboembolic (TE) events preceding cancer have been observed. Some studies failed to find this correlation. We retrospectively examined the cancer incidence following thromboembolic events in patients at our medical center. Medical records of 183 patients with established thromboembolic events documented in their records were selected and reviewed. Time interval between primary, secondary, and recurrent TE events preceding cancer diagnosis was analyzed. Two hundred age- and sex-matched controls seen during the same period and without any evidence of TE were randomly selected and charts reviewed for malignancy. Cancer occurred after TE in 48 of 183 patients (26.2%). In controls, cancer was diagnosed in 23 (11.5%). This was statistically significant with an odds ratio of 2.736 (1.586, 4.720). In the 64 primary TE patients, the cancer incidence was 37.5%. The 63 patients with recurrent TE had an incidence of 35.4%, and 56 secondary TE patients had an incidence of 27.1%. Time between initial TE and cancer diagnosis was <6 months in 27 (56.3%) patients, between 6 months and 1 year in 12 (25.0%), 1-5 years in 5 (10.4%), and >5 years in 4 (8.3%). Fourteen (31.1%) TE patients presented with metastatic cancer. This study indicates that thromboembolic events are important predictors of cancer. Cancer in this population occurs within a year in the majority of patients. Cancer screening in patients without identifiable risk factors for thrombosis could be helpful for early detection, diagnosis, and management of cancer. PMID- 12555215 TI - Thromboxane synthesis is increased by upregulation of cytosolic phospholipase A2 and cyclooxygenase-2 in peripheral polymorphonuclear leukocytes during bacterial infection in childhood. AB - Prostaglandins (PGs) and thromboxane (TX) are important mediators of inflammation. Recent studies revealed that PG and TX synthesis is controlled by the regulation of PG- and TX-synthesizing enzymes. In this study, we examined the TX synthesis and the expression of TX-synthesizing enzymes in activated peripheral polymorphonuclear leukocytes (PMNs) obtained from children with bacterial infection. Blood samples were obtained from controls and patients with bacterial infection. A23187-stimulated production of TXB(2), a stable metabolite of TXA(2) in PMNs, was measured by a specific radioimmunoassay. The mRNA expression of cytosolic phospholipase A(2) (cPLA(2)), cyclooxygenase (COX)-1, COX 2, and TXA(2) synthase was determined by RT-PCR. The synthesis of TXB(2) in PMNs was significantly increased in the patients [925.0 (550.0-1100.0) pg/10(6) cells], compared with the controls [550.0 (450.0-775.0) pg/10(6) cells]. The mRNA expression for cPLA(2) and COX-2 in PMNs was also enhanced in the patients. The results indicate that TX production in PMNs is significantly increased through possible transcriptional mechanisms of cPLA(2) and COX-2 during bacterial infection in children. The upregulation of TXA(2) synthesis may contribute to the process of acute inflammatory reaction caused by bacterial infection. PMID- 12555216 TI - Concordant fetal hemoglobin response to hydroxyurea in siblings with sickle cell disease. AB - Fetal hemoglobin (HbF) level and the HbF responses to hydroxyurea (HU) vary among patients with sickle cell disease and are, at least in part, genetically regulated. We hypothesized that siblings with sickle cell disease are likely to share the same parental beta-like globin gene clusters with their cis-acting regulatory sequences and therefore, if regulation of this response is linked to the beta-globin gene cluster, might have concordant HbF responses to HU. Accordingly, we studied 26 families (30 sib pairings), 20 with sickle cell anemia (three families had three siblings) and 6 families with HbS-beta-thalassemia (one family had three siblings, and one family consisted of monozygotic twins), to see if siblings with sickle cell disease had discordant or concordant changes in HbF during HU treatment. Intraclass correlation coefficients (r) showed a high, positive correlation between sibs for HbF levels before and during HU treatment and a concordant change in HbF response from baseline to treatment-associated levels. Changes in mean corpuscular volume (MCV) paralleled HbF levels, while the expected correlations between treatment-associated fall in leukocyte count and increase in MCV were also present. Our results provide additional evidence that some elements that regulate HbF expression are linked to the beta-globin gene cluster. PMID- 12555217 TI - Ethnicity and prognosis in acute myeloid leukemia. AB - Ethnicity has been described as a prognostic factor in breast cancer and in childhood acute lymphocytic leukemia but not in adult acute myeloid leukemia (AML). We reviewed the records of 225 consecutive AML patients who were diagnosed and treated between 1983 and 1995. Data were collected concerning demographic factors, presenting clinical features, and treatment protocols. We categorized ethnicity as follows: European Jews, non-European Jews, and Arabs. We assessed the role of ethnicity controlling for other known prognostic factors on treatment outcome and survival in this population. Older age, high leukocyte count at diagnosis, and high-risk chromosomal aberrations were significantly associated with overall survival in univariate analysis. In multivariate analysis high leukocyte count and high-risk chromosomal aberrations exerted an independent negative effect on survival. European origin was associated with longer event free survival in univariate analysis (P = 0.024) and longer overall (P < 0.01) and event-free (P < 0.01) survival but not with a higher remission rate in multivariate analysis. For AML patients who achieved remission after induction chemotherapy and survived its complications, European origin is an independent favorable prognostic factor for long-term remission and survival in Israel. These findings may reflect better socioeconomic status, social support, increased compliance with treatment protocols, or better psychological coping mechanisms with malignancy. PMID- 12555218 TI - Acute myelogenous leukemia with the t(3;12)(q26;p13) translocation: case report and review of the literature. AB - Translocations involving the EVI1/MDS1 gene at 3q26 and the TEL gene at 12p13 are comparatively common in acute leukemia, but a translocation between the two genes has been reported only in a handful of cases. We report an additional case of acute myelogenous leukemia (AML) preceded by a myelodysplastic syndrome (MDS) with the translocation t(3;12)(q26;p13) in a 36-year-old woman. The translocation was present early in the disease and long before the MDS progressed to AML 3 years after diagnosis. At the time of progression to AML, an additional chromosomal abnormality, a monosomy 22, was discovered. The patient was treated with the protocol MAQ, which comprised mitoxantrone, aracytine, and quinine, as her blasts expressed the p-glycoprotein, but she failed to obtain remission. A second treatment with the same protocol resulted in only minimal response. The patient was treated again with high-dose Ara-C and idarubicine in an attempt to achieve a response before allogeneic unrelated transplantation, but she did not respond to the treatment and died shortly thereafter. A review of the literature revealed 12 other cases of the t(3;12)(q26;p13) translocation. Characteristics of those cases are reviewed in this article. PMID- 12555219 TI - Monocytic leukemia with CALM/AF10 rearrangement showing mediastinal emphysema. AB - The t(10;11)(p12-14;q14-21) is a non-random translocation that results in the fusion of CALM gene on chromosome 11 with AF10 gene on chromosome 10. This translocation is observed in acute myeloid leukemia, acute lymphoblastic leukemia, and lymphoblastic lymphoma. Here we report a patient with t(10;11) who was diagnosed with AML-M4. Reverse transcriptase-polymerase chain reaction (RT PCR) assay revealed one type of CALM/AF10 and three types of AF10/CALM fusion transcripts. Sequencing analysis for these RT-PCR products determined the breakpoint in CALM at nucleotide (nt) 1926-1927 and in AF10 at nt 423-424. The latter breakpoint was the same as that identified in three monocytic cell lines carrying t(10;11). After achieving complete remission, the patient developed mediastinal emphysema during the course of consolidation therapy, possibly due to the necrosis of his mediastinal mass. Monocytic leukemias with CALM/AF10 fusion are frequently associated with mediastinal invasion. We need to pay special attention to such a complication, even if the chest X-ray is normal at presentation. PMID- 12555220 TI - Successful engraftment and stable full donor chimerism after myeloablation with thiotepa, fludarabine, and melphalan and CD34-selected peripheral allogeneic stem cell transplantation in hemophagocytic lymphohistiocytosis. AB - Allogeneic hematopoietic stem cell transplantation (HSCT) represents the only curative option for primary hemophagocytic lymphohistiocytosis (HLH), a rare disease of infants and young children, characterized by recurrent fever, hepatosplenomegaly, and cytopenia. We report a case of successful engraftment and stable full-donor chimerism in a patient with HLH who underwent peripheral allogeneic CD34-selected HSCT. The donor was his 1-antigen-HLA-mismatched grandmother. After a conditioning regimen based on the combination of thiotepa, fludarabine, melphalan, and rabbit antilymphocyte serum, the patient received a megadose of 26.3 x 10(6)/kg of CD34(+) peripheral blood cells. Neutrophil (>0.5 x 10(9)/L) and platelet (>50 x 10(9)/L) engraftment was observed on days +16 and +12, respectively, and the patient was discharged home on day +24. No acute or chronic GVHD was observed. Infectious complications were the main causes of re hospitalization in the first year after transplantation, but no significant morbidity was observed thereafter. Thirty-two months after HSCT, the patient is alive and well, still in complete clinical remission of his underlying disease with a durable engraftment, normal NK activity and full donor chimerism. This case suggests that a fludarabine-based conditioning regimen and CD34-selected peripheral allogeneic HSCT may be a feasible option in case of unavailability of a fully HLA-matched related or unrelated donor. PMID- 12555221 TI - CTLA-4 exon 1 polymorphism in patients with autoimmune blood disorders. AB - CTLA-4 is a CD28 homologue that plays an important role in negative regulation of T-cell responses. Its transient expression on the surface of activated T cells antagonizes the activating signals and terminates the T-cell response. An A to G polymorphism at position 49 of the CTLA-4 first exon has recently been associated with several autoimmune disorders. In the present study we have examined the prevalence of the A and G alleles of the CTLA-4 gene in 50 patients with autoimmune hemolytic anemia (AIHA), of which 20 had idiopathic AIHA and 30 had AIHA and chronic lymphocytic leukemia (CLL), and in 60 patients with immune thrombocytopenic purpura (ITP). Control subjects were 100 healthy individuals and 100 CLL patients without clinical evidence for an autoimmune disease. The G allele was present at a significantly higher frequency among the patients with AIHA (P = 0.003), whereas no difference was observed between patients with ITP and controls. The G allele frequency was highest among CLL patients who had developed AIHA. The obtained data indicate that the G allele of CTLA-4 predisposes to the development of AIHA, particularly among patients with CLL. PMID- 12555222 TI - G6PD deficiency and malaria in India. PMID- 12555223 TI - Hyperhomocysteinemia in healthy Asian Indians. PMID- 12555224 TI - Essential thrombocytosis and liver transplantation. PMID- 12555225 TI - Spontaneous regression of HIV-associated Hodgkin's disease. PMID- 12555226 TI - Unusual case of antiphospholipid antibody syndrome presenting with extensive cutaneous infarcts in a patient on long-term procainamide therapy. PMID- 12555227 TI - Association of a MAOA gene variant with generalized anxiety disorder, but not with panic disorder or major depression. AB - This study was conducted to detect a possible association of a T941G single nucleotide polymorphism (SNP) in the monoamine oxidase A (MAOA) gene with generalized anxiety disorder (GAD), panic disorder (PD), or major depression (MD). Fifty GAD patients (34 females and 16 males), 38 PD patients (21 females and 17 males), and 108 MD patients (80 females and 28 males) were included. The comparison group consisted of 276 (132 females and 144 males) unrelated healthy individuals. The 941T allele was over-represented in patients suffering from GAD (chi(2) = 6.757; df = 1; P < 0.01, not corrected for multiple testing) when compared to healthy volunteers. No association was observed in MD or PD. This is the first study specifically analyzing the MAOA G941T polymorphism in GAD and thus needs to be replicated in an independent sample. However, the results are in line with previous data suggesting an association between the MAOA locus and regulation of complex human behavior. PMID- 12555228 TI - Role of elastin polymorphisms in panic disorder. AB - Panic disorder (PD) is a complex neuropsychiatric disorder that has been associated with an increased frequency of mitral valve prolapse. Elastin is a prominent component of mitral valves and, in a genome screen of 23 pedigrees with PD, we found evidence of linkage to the region of chromosome 7 that contains the elastin gene (ELN). Therefore, we examined the minimal essential promoter and coding regions of ELN in 23 independent probands from the families in our linkage studies using single strand conformational polymorphism analysis. We found three polymorphisms including one that coded for a non-conservative amino acid change. However, none of these polymorphisms were associated with panic disorder in a case-control analysis or linked to it in multiplex pedigrees. In our pedigrees, exonic polymorphisms in ELN do not play a major role in the genetic vulnerability to PD. PMID- 12555229 TI - Association of an exonic LDHA polymorphism with altered respiratory response in probands at high risk for panic disorder. AB - Panic disorder (PD) is a clinical syndrome characterized by recurrent discrete episodes of fear accompanied by a variety of physiological and psychological symptoms, often with prominent respiratory components. A series of clinical observations has led some investigators to hypothesize that subtle alterations in ventilatory regulation are integral to at least a subtype of PD. In order to identify genetic factors that might predispose individuals to these alterations in ventilatory response, we conducted single stranded conformation polymorphism analysis across the exons of the lactate dehydrogenase A and B genes (LDHA and LDHB) using DNA prepared from 86 subjects previously characterized by respiratory response to a CO(2) challenge with a variable genetic loading for PD. Remarkably, a single conserved LDHA exon 5 haplotype conferred increased risk for a paradoxical ventilatory response pattern to CO(2) inhalation which robustly separated well subjects at high risk for PD from low-risk control subjects. But, comparison of LDHA exon 5 genotypes in PD probands (n = 25) to that of random newborn controls (n = 182) did not demonstrate any significant differences. Given the pivotal role of LDH in the metabolism of lactate, a known inducer of panic attacks, and the dependence of LDH activity on cell pH, we suggest that LDHA polymorphisms may contribute to the variability to CO(2) respiratory challenge. PMID- 12555230 TI - Possible parent-of-origin effect of Dopa decarboxylase in susceptibility to bipolar affective disorder. AB - Dopa decarboxylase (DDC) catalyses the synthesis of both dopamine and serotonin as well as trace amines suggested to possess neuromodulating capabilities. We have previously reported evidence suggesting an association between DDC and bipolar affective disorder (BPAD) [Borglum et al., 1999]. To further investigate the possible role of DDC in BPAD, we analyzed a 1- and a 4-bp deletion variant both of putative functional significance-in two new samples: a case-control sample with 140 cases and 204 controls, and 100 case-parents trios. We also tested for association in subjects with familial disease in both the new and the previously investigated samples. The previously reported association was not replicated in either of the new samples. However, a preponderance of the 1-bp deletion was increased by analysis of the familial cases separately for all case control samples investigated, indicating a possible association with familial disease (combined analysis, P = 0.02). In the trio sample, a preferential paternal transmission of the 4-bp deletion was observed (P = 0.006). DDC is located next to the imprinted gene GRB10, which is expressed specifically from the paternal allele in fetal brains. Increased transmission of paternal DDC alleles has also been suggested in attention deficit hyperactivity disorder. We suggest that DDC might confer susceptibility to BPAD predominantly when paternally transmitted. PMID- 12555231 TI - Genetic refinement and physical mapping of a 2.3 Mb probable disease region associated with a bipolar affective disorder susceptibility locus on chromosome 4q35. AB - A susceptibility locus for bipolar affective disorder has been mapped to chromosome 4q35 in a large multigenerational pedigree. We have expanded this analysis to include 55 pedigrees (674 individuals, 214 affecteds). The evidence for linkage to 4q35 was strengthened in this larger cohort, with a maximum two point LOD score of 3.2 for marker D4S1652. Several other markers in the region gave LOD scores greater than 1.5. Non-parametric analysis provided additional support for linkage to the 4q35 region. To further refine this region, haplotype analysis was carried out in 16 of the 55 pedigrees that showed evidence of linkage. As there is no evidence for an ancestral haplotype, nor a one-to-one correspondence between the disease and putative disease haplotype, we undertook an analysis based on pedigree-specific, identical-by-descent allele-sharing in order to define a probable disease region. This analysis indicated that the percentage sharing of alleles, identical-by-descent, in affecteds of all linked pedigrees increases from 60% at the centromeric markers to 75% for markers at the telomere. Maximal allele sharing occurred between markers D4S3051 and 4qTEL13 with this 24 cM region defining a probable disease region. We have constructed a physical map of the 4q35 interval consisting of a YAC contig and BAC clones. Based on this map the probable disease region between D4S3051 and 4qTEL13 corresponds to only 2.3 Mb. This region is very gene poor with only three known genes indicated from the YAC/BAC map. The small number of genes will facilitate systematic screening for variations associated with bipolar disorder. PMID- 12555232 TI - Dopamine beta-hydroxylase (DBH) gene and schizophrenia phenotypic variability: a genetic association study. AB - Recently, two polymorphisms (DBH5'-Ins/del and DBH 444 g/a) of the Dopamine Beta Hydroxylase (DBH) gene were isolated, and one haplotype (Del-a) was found to be associated with low DBH activity and cocaine-induced paranoia. The purpose of this study is to test for association between these two polymorphisms and schizophrenia or its phenotypic variability with respect to neuroleptic therapeutic response and symptom profile. Allelic and haplotype distributions of these two polymorphisms were compared between two groups of schizophrenic patients (excellent neuroleptic-responders; R, n = 42 and non-responders; NR, n = 64), and one group of healthy volunteers (n = 120). The "Del" and "a" alleles were in positive linkage disequilibrium. No allelic or genotype differences in the distribution of these two polymorphisms were observed between patients and controls. However, The Del-a haplotype was significantly more common in NR patients, and the mean total BPRS score was significantly higher in the group of patients with the Del-a compared to those without the Del-a haplotype. These results suggest that the DBH gene is not a causative factor in schizophrenia but that it may be a modulator of psychotic symptoms, severity of the disorder and therapeutic response to neuroleptic drugs. PMID- 12555233 TI - Association of GABA(A) receptors and alcohol dependence and the effects of genetic imprinting. AB - GABA receptor genes have been postulated as candidates affecting the risk for alcoholism. The potential association between genes encoding five subunits of the GABA(A) receptors and alcoholism (alcohol dependence) was analyzed in the multiplex alcoholic pedigrees collected by the Collaborative Study on the Genetics of Alcoholism (COGA) using family-based association tests. We found consistent, although weak, linkage disequilibrium between GABRB1 (located on chromosome 4) and alcoholism (P < 0.03). Genes encoding GABRA1 and GABRA6, on chromosome 5, did not provide evidence for association with alcoholism. GABRA5 and GABRB3, on chromosome 15, were reported to be expressed uniparentally from the paternal chromosome. Analyses of paternal transmission of alleles of GABRA5 provided evidence for association with alcoholism, particularly in the Caucasian population and with the stricter ICD-10 definition of alcoholism (P < 0.004). Evidence of association was also observed during paternal transmission with GABRB3 in the Caucasian population (P < 0.007). Maternal transmissions provided no evidence for association. These data are consistent with an association between the expressed alleles in the GABA(A)-gene cluster on chromosome 15 and alcoholism that is modulated by genetic imprinting. PMID- 12555234 TI - Functional variation in promoter region of monoamine oxidase A and subtypes of alcoholism: haplotype analysis. AB - Monoamine oxidase (MAO) is a mitochondrial enzyme involved in the degradation of certain neurotransmitter amines. MAO-A, due to its function in central nervous system, has been one of the important candidate genes involved in the development of neuropsychiatric disorders. A functional polymorphism in the promoter region of the MAO-A gene has been identified. This variation affects the transcriptional efficiency of the gene. To determine the role of this MAO-A functional polymorphism in the development of subtypes of alcoholism, a sample of alcoholics and normal controls were screened with this marker. The allele frequency differences between total alcoholics, Types I and II alcoholics, and normal controls was not significant. Comparison of male alcoholics to male normal controls for the frequencies of two-loci and three-loci haplotypes was statistically significant. After Bonferroni's correction for multiple tests none of the results remained significant at P < 0.05. Our results indicate that MAO-A may play a role in the development of alcoholism but the gene effect is very small. PMID- 12555235 TI - Association of EEG coherence and an exonic GABA(B)R1 gene polymorphism. AB - The GABA(B) receptor 1 gene is mapped to chromosome 6p21.3 within the HLA class I region close to the HLA-F gene. Susceptibility loci for epilepsy and schizophrenia have been mapped in this region. Based on pharmacological evidence, it has been suggested that GABA(B) receptors may play a crucial role in the synchronization of EEG oscillations, which in turn can be abnormal in neuropsychiatric disorders. In the present study, the hypothesis was tested, whether three exonic variants of the gene encoding the human GABA(B) receptor (GABA(B)R1) modify cortical synchronization measured as scalp-recorded EEG coherence. Two principal components of EEG coherence (frontal coherence, parietotemporal coherence) were investigated in 104 healthy subjects during three conditions: resting EEG, activated EEG, and event-related EEG. No significant associations were found between the frontal coherence component and any polymorphism or between the parietotemporal coherence component and the exon 1a1 polymorphism. However, parietotemporal coherence showed statistically highly significant associations across all three experimental conditions with exon 7 and trend associations with exon 11. The results provide evidence that the translated polymorphism of exon 7 may be functionally meaningful and impact cortical EEG oscillations. Since variations of EEG coherence have been described for several neuropsychiatric disorders, the present association should be tested in clinical samples using EEG coherence as an intermediate phenotype. PMID- 12555236 TI - Relationship between functional dopamine D2 and D3 receptors gene polymorphisms and neuroleptic malignant syndrome. AB - Our previous study has suggested that the TaqI A polymorphism of dopamine D2 receptor gene (DRD2) is associated with the predisposition to neuroleptic malignant syndrome (NMS). However, the specificity of this polymorphism as a predictor of NMS dose not seem to be sufficient enough. Meanwhile, it has been shown that the non-carriers of Del allele of the -141C Ins/Del polymorphism in the promoter region of DRD2 have lower dopamine D2 receptor in the brain than the carriers. In addition, dopamine D3 receptor gene has a Ser(9)Gly polymorphism, which may alter the receptor function. The present study examined the association between these three polymorphisms and the development of NMS to investigate if a combination of these polymorphisms could increase the specificity as markers for NMS. The subjects were 17 psychiatric patients who had developed NMS (13 patients with schizophrenia, 3 with major depression, and 1 with dementia of the Alzheimer's type) and 163 schizophrenic patients who had never developed this syndrome. The frequency of the A1 allele was significantly (P = 0.012) higher in the patients who had developed NMS (59%) than in the patients who had not (35%). The proportion of the A1 carriers was significantly (P = 0.003) higher in the patients with NMS (16/17: 94%) than in those without the syndrome (93/163: 57%). However, no significant differences were found in the allele and genotype frequencies of the other two polymorphisms between the two groups. The present study suggests that only the TaqI A polymorphism is at least partly useful as a predictor of NMS, but the -141 C Ins/Del and Ser(9)Gly polymorphisms are not. PMID- 12555237 TI - Association study between dopamine D3 receptor gene variant and personality traits. AB - Dopamine receptor gene variation has been hypothesized to influence personality traits characterized by novelty seeking and related traits. We analyzed a dopamine D(3) receptor gene (DRD3) variant in a Swedish population (n = 373) investigated with one or more of several personality questionnaires. No significant relationships were found between DRD3 genotypes and any of the 15 Karolinska Scales of Personality (KSP) and five Health-relevant Personality 5 factor inventory (HP5i) scales. The DRD3 variant was associated with some scales related to novelty seeking: the Swedish universities Scales of Personality (SSP) Adventure Seeking and the revised NEO personality inventory (NEO-PI-R) Fantasy (O1) and Order (C2) scales. There were also associations with the Temperament and Character Inventory (TCI) Cooperativeness and Compassion (C4) scales. After correction for multiple testing, however, no significant difference remained. We conclude that the investigated DRD3 polymorphism does not have a major impact on personality in the investigated population. PMID- 12555238 TI - Tridimensional personality questionnaire trait of harm avoidance (anxiety proneness) is linked to a locus on chromosome 8p21. AB - Human personality traits are moderately heritable but only recently have specific polymorphisms been associated with particular personality dimensions especially anxiety-related and novelty-seeking traits. The first genome-wide scan for personality traits was recently carried out by Cloninger et al. [1998: Am J Med Genet 81:313-317] and his colleagues and they reported that a region on 8p21 showed linkage to TPQ Harm Avoidance, an anxiety-related personality trait. Towards replicating and extending these results, we examined both 8p21 and two additional chromosomal regions (1q21-24 and 22q12-13) for linkage to TPQ personality traits by genotyping at least three microsatellite markers in each region in a group of 384 sibling pairs. We found evidence for linkage to TPQ HA at 8p21-23 (Lod score = 2.907) confirming in an independent sample the initial findings by Cloninger and his colleagues. PMID- 12555239 TI - Human nuclear transcription factor gene CREM: genomic organization, mutation screening, and association analysis in panic disorder. AB - Panic disorder is an anxiety disorder with an estimated heritability of 48%. Variation in the gene of the nuclear transcription factor "cAMP-responsive element modulator" (CREM) might contribute to its pathogenesis. CREM knock-out mice exhibit significantly less anxiety behavior than wild-type mice and the alternative CREM gene product "inducible cAMP early repressor" (ICER) plays a pivotal role in the hypothalamo-pituitary-adrenal (HPA) axis, which is disturbed in panic disorder. We characterized the genomic organization of the human CREM gene and performed a systematic mutation screening by means of single stranded conformational analysis (SSCA) in a sample of 40 German patients with panic disorder (DSM-III-R). Four novel single nucleotide polymorphisms in CREM promoters P 1 and P 4, one trinucleotide (ATT)-repeat polymorphism in CREM promoter P 2-generating the ICER isoform-and a rare amino acid substitution in CREM exon glut 2 were identified. Association analysis in an extended sample of German patients (n = 88) revealed a significant excess of the shorter CREM P 2 promoter eight-repeat trinucleotide allele and of genotypes containing the eight repeat trinucleotide allele in panic disorder (P = 0.02), in particular in panic disorder without agoraphobia (P = 0.001). A replication study in independent Italian (n = 76) and Spanish (n = 62) samples, however, failed to confirm this observation. This suggests that the CREM P 2 promoter trinucleotide polymorphism is not a major susceptibility factor in the pathogenesis of panic disorder. Functional analysis of the observed CREM P 2 promoter polymorphism as well as studies in independent panic disorder samples are necessary. PMID- 12555240 TI - Testing the test--why pursue a better test for Huntington disease? AB - In 1993, the gene mutation for Huntington disease (HD) was identified and testing became possible with a reliability of >99%, without the need for co-operation of relatives. In 1997, a systematic information program offered the mutation retest to individuals who had earlier received a linkage test result for HD, which has a residual uncertainty of 1-9%. The characteristics of 129 individuals tested by linkage analysis for HD are reported on, as well as the reasons for their reassessment by mutation testing. Three groups were compared: (1) people who were retested between 1993 and 1997, before this study had started, (2) people who were retested after we provided information, and (3) persons who refrained from retesting. Nearly half of the linkage-tested individuals were retested, with the exception of noncarriers with a residual risk of 1 or 2%. Of them, less than one out of five were retested. Carriers with a hopeful view on the future (BHS) and a better sense of well-being (GHQ) were more likely to have the retest. Female carriers were also more likely to have the retest before we contacted them. Noncarriers who were retested were more anxious (HADS) than noncarriers who refrained from the retest. Retestees were younger at the time of testing. No risk reversals were revealed by this study. PMID- 12555241 TI - An investigation of the 5-HT2C receptor gene as a migraine candidate gene. AB - Migraine is a common complex disorder, currently classified into two main subtypes, migraine with aura (MA) and migraine without aura (MO). The strong preponderance of females to males suggests an X-linked genetic component. Recent studies have identified an X chromosomal susceptibility region (Xq24-q28) in two typical migraine pedigrees. This region harbours a potential candidate gene for the disorder, the serotonin receptor 2C (5-HT(2C)) gene. This study involved a linkage and association approach to investigate two single nucleotide variants in the 5-HT(2C) gene. In addition, exonic coding regions of the 5-HT(2C) gene were also sequenced for mutations in X-linked migraine pedigrees. Results of this study did not detect any linkage or association, and no disease causing mutations were identified. Hence, results for this study do not support a significant role of the 5-HT(2C) gene in migraine predisposition. PMID- 12555242 TI - Defining the autism minimum candidate gene region on chromosome 7. AB - Previous genetic and cytogenetic studies provide evidence that points to one or more autism susceptibility genes residing on chromosome 7q (AUTS1, 115-149 cM on the Marshfield map). However, further localization using linkage analysis has proven difficult. To overcome this problem, we examined the Collaborative Linkage Study of Autism (CLSA) data-set to identify only the families potentially linked to chromosome 7. Out of 94, 47 families were identified and 17 markers were used to generate chromosomal haplotypes. We performed recombination breakpoint analysis to determine if any portion of the chromosome was predominately shared across families. The most commonly shared region spanned a 6 cM interval between D7S501 and D7S2847. Additional markers at 1 cM intervals within this region were genotyped and association and recombination breakpoint analysis was again performed. Although no significant allelic association was found, the recombination breakpoint data points to a shared region between D7S496-D7S2418 (120-123 cM) encompassing about 4.5 Mb of genomic DNA containing over 50 genes. PMID- 12555243 TI - Absence of MeCP2 mutations in patients from the South Carolina autism project. AB - The methyl-CpG binding protein 2 (MeCP2) gene has recently been identified as the gene responsible for Rett syndrome (RS), a pervasive developmental disorder considered by many to be one of the autism spectrum disorders. Most female patients with MeCP2 mutations exhibit the classic features of RS, including autistic behaviors. Most male patients with MeCP2 mutations exhibit moderate to severe developmental delay/mental retardation. Ninety nine patients from the South Carolina autism project (SCAP) were screened for MeCP2 mutations, including all 41 female patients from whom DNA samples were available plus the 58 male patients with the lowest scores on standard IQ tests and/or the Vineland Adaptive Behavior Scale. No pathogenic mutations were observed in these patients. One patient had the C582T variant, previously reported in the unaffected father of an RS patient. Two other patients had single nucleotide polymorphisms in the 3' UTR of the gene, G1470A and C1516G. These variants were seen in 12/82 and 1/178 phenotypically normal male controls, respectively. The findings from this and other studies suggest that mutations in the coding sequence of the MeCP2 gene are not a significant etiological factor in autism. PMID- 12555244 TI - Heritability of attention problems in children: I. cross-sectional results from a study of twins, age 3-12 years. AB - Multiple twin studies of attention problems (AP) from the Child Behavior Checklist or ADHD from the DSM criteria have reported on the genetic and environmental influences on these behaviors. The majority of these have studied AP and ADHD symptoms in twin samples combined across wide age spans, combined rater information and both genders. Thus, it is possible that the results are complicated by developmental, informant, and gender differences. The purpose of this study was to assess for the genetic and environmental contributions to overactive behavior (a syndrome highly related to AP in 7-, 10-, and 12-years olds) in 3-years olds (3,671 twin pairs), and attention problems in 7- (3,373 twin pairs), 10- (2,485 twin pairs), and 12-years olds (1,305 twin pairs) while controlling for developmental, gender and rater contrast contributions. Using a cross-sectional twin design, contributions from genetic additive, genetic dominance, unique environmental and rater contrast effects were estimated for CBCL maternal reports. We found that genetic influences on overactive behavior and attention problems are high across an age span that covers pre-school and elementary school age. Although girls display less problem behavior compared to boys, heritability estimates were found equal for both genders at each age. Environmental experiences that are unique to the individual accounted for the remaining influence. At the age of 3 years, a rater contrast effect was detected. We hypothesize that the contrast effect represents a maternal rater bias effect that is dependent on the age of the twins. The implications of these findings are discussed with reference to the clinical setting and in the context of future research. PMID- 12555245 TI - Association of the 3' UTR transcription factor LBP-1c/CP2/LSF polymorphism with late-onset Alzheimer's disease. AB - Alzheimer's disease (AD) is a genetically heterogeneous neurodegenerative disorder. To date, apolipoprotein E (apoE) is the only established susceptibility gene for late-onset AD. ApoE accounts for less than 50% of the risk of AD, indicating the presence of other unknown susceptibility loci. Linkage studies have indicated chromosome 12 as the most likely location for another late-onset AD locus. We examined seven polymorphisms in five candidate genes located in and around the linkage peaks on chromosome 12 in 564 cases and 523 controls. The genes included complement component 1R (C1R), vitamin D receptor (VDR), scavenger receptor B1 (SR-B1), low-density lipoprotein receptor related protein 1 (LRP1), and transcription factor LBP-1c/CP2/LSF. We found no association with C1R, VDR, SR-B1, and LRP1 polymorphisms. However, the frequency of the A allele of the 3' (untranslated region) UTR LBP-1c/CP2/LSF polymorphism was higher in controls than cases (0.071 vs. 0.051; P = 0.042) with an adjusted odds ratio (OR) of 0.65 (95% confidence interval [CI]: 0.43-0.96; P = 0.0498). Our data suggest that the LBP 1c/CP2/LSF polymorphism may have a moderate protective effect against the risk of AD. PMID- 12555246 TI - Effects of retinoic acid on proliferation, apoptosis, cytotoxicity, migration, and invasion of neuroblastoma cells. AB - BACKGROUND: Because of the known property of less aggressiveness of differentiated cells compared to immatured cells all attempts are made to elucidate whether differentiation inducers possibly could be applied for neuroblastoma therapy. We are interested in examining the influence of retinoic acid (RA) on proliferation, apoptosis, cytotoxicity, migration, and invasion in dependence of the differentiation of neuroblastoma cells classified into N-type (SK-N-FI, SH-SY5Y), I-type (SK-PN-DW), and S-type (SK-N-LO, SK-N-MC) cells. PROCEDURE: Neuroblastoma cells were exposed to 10(-5) M RA and 200 ng/ml camptothecin (CAM) (control substance for apoptosis). Proliferation, apoptosis, and cytotoxicity were quantified by photometric assays. The influence on migration and invasion of neuroblastoma cells was examined by a scratch-test and by the measurement of the invasion through matrigel coated chamber inserts. RESULTS: In general, RA treatment induced proliferation inhibition predominantly in the cell lines SK-PN-DW (16%, P < 0.05) and SK-N-MC (8%, (P < 0.001), respectively. In the N-type cell lines SK-N-FI (P > 0.05) and SH-SY5Y (P < 0.001) no proliferation inhibition was determined conforming with no detection of apoptosis. CAM confirmed its capability to induce apoptosis in the cell lines SH SY5Y (43.6%, P < 0.05), SK-PN-DW (54.8%, P > 0.05), and SK-N-MC (28.9%, P < 0.0 01) except for SK-N-FI with only 9.3% (P > 0.05), but after 24 hr of treatment. Minor signs of restricted migration were observed, while RA treatment reduced significantly the invasion rate through Matrigel of SK-N-FI to 13.3% (P < 0.01), SH-SY5Y to 19.2% (P < 0.05), SK-N-MC to 27.8% (P < 0.05), and SK-N-LO to 17.7% (P < 0.01). CONCLUSIONS: It is demonstrated that RA treatment can interfere with cell growth and in invasion by inducing neuronal differentiation in N-type and apoptosis in S-type neuroblastoma cell lines. PMID- 12555247 TI - Clinical outcome in children with craniopharyngioma treated with primary surgery and radiotherapy deferred until relapse. AB - BACKGROUND: To report the clinical outcome in children with craniopharyngioma following primary surgery and deferred radiotherapy at relapse. PROCEDURE: Twenty five children with craniopharyngioma were treated with primary surgery. Total resection was achieved in 19 children (76%), while in 24% total resection was not achieved due to tumor adhesion to adjacent critical structures. None of these children received radiation therapy immediately after total or sub-total resection. Radiotherapy was delivered at the time of relapse in 11 patients (44%). RESULTS: The median follow-up from primary surgery was 10 years (3-16 years). The 14 year overall survival was 100%. Tumor recurrence was observed in (12/25) 48% at a median interval of 17 months. Tumor recurrence following total resection was 6/19 (32%) compared to 100% (6/6) following sub-total resection, and radiotherapy. The 2, 3, and 6 years relapse-free survival following initial surgery was 72, 55, and 50%, respectively. Univariate analysis revealed only extent of surgery to be significant for local recurrence (P < 0.0001). The sequelae observed in these patients included panhypopituitarism (100%), appetite disorders and hypothalamic obesity (32%), neuropsychological and behavioral disorders (20%), and sleep disorders (12%). Majority of children with non endocrine complications had a local recurrence requiring further surgery and radiotherapy. CONCLUSIONS: The two standard treatment options in children with craniopharyngioma are primary surgery and sub-total resection followed by radiotherapy. In certain subgroups of patients such as those with large tumors and hypothalamic extension, primary surgery is associated with a high incidence of complications and high failure rates. We recommend utilization of an individualized risk-based treatment approach, that attempts to maximize cure rates without compromising long-term functional outcome in children with craniopharyngiomas. PMID- 12555248 TI - Human polyomavirus DNA is not detected in Guthrie cards (dried blood spots) from children who developed acute lymphoblastic leukemia. AB - BACKGROUND: Epidemiological evidence has suggested that some childhood acute lymphoblastic leukemia (ALL) may be initiated in utero and may have an infectious etiology. The human polyomavirus JC virus (JCV) has been discussed as a candidate virus, but its presence has not been demonstrated in leukemia cells from children with ALL. The aim of this study was, therefore, to investigate if prenatal human polyomavirus infection could still indirectly be correlated to the development of childhood ALL. PROCEDURE: Fifty-four Guthrie cards (stored, dried blood spots filter papers, routinely collected from newborns for different screening analyses), collected at 3-5 days of age, from Swedish children who subsequently developed ALL, as well as from 37 healthy controls, were investigated by nested PCR for the presence of human polyomaviruses JCV and BK virus (BKV). RESULTS: JCV and BKV DNA were not detected in any of the Guthrie cards from ALL patients or from healthy controls, although all tested samples had amplifiable DNA as confirmed by an HLA DQ PCR. CONCLUSIONS: JCV or BKV were not found in any of the dried blood spots of children who later developed ALL or in the healthy controls. These findings suggest that it is unlikely that childhood ALL is associated with an in utero infection with JCV or BKV, although it is not possible to exclude an association with an in utero infection that has become latent in the kidneys with very low levels of circulating virus at birth. PMID- 12555249 TI - Hypothalamo-pituitary-adrenal axis integrity after cranial irradiation for childhood posterior fossa tumours. AB - BACKGROUND: The evolution of anterior pituitary deficits after treatment for pituitary tumours has been largely attributed to local irradiation, but may be influenced as much by tumour mass or surgery. Other than growth hormone (GH) insufficiency, the late endocrinopathies after survival from non-central brain tumours have been little documented. The aim of this study was to investigate the hypothalamic-pituitary-adrenal (HPA) axis in long-term survivors of cranial irradiation for childhood posterior fossa tumours. PROCEDURE: We studied long term data in patients treated prepubertally for posterior fossa brain tumours and systematically referred by radiation oncologists for growth and pubertal monitoring to the London Centre for Paediatric Endocrinology over the last 25 years. They must have undergone HPA axis assessment twice, first prepubertally at documentation of growth failure, and second at completion of growth and puberty. Data on sixteen patients (12 males, 4 females; median age: 5.7 years, range: 2.5 8.8 years), who had undergone excision surgery with high dose cranial irradiation and/or chemotherapy for childhood posterior fossa tumours, were examined. Patients were followed for a median of 11.0 (range: 6.8-21.4) years after radiotherapy. HPA axis assessment was undertaken with the insulin-induced hypoglycaemia test (ITT). Basal thyroid, cortisol and gonadal function tests were undertaken annually throughout the follow-up period and any deficits replaced. RESULTS: At each ITT, all patients mounted an inadequate GH response. By the end of the follow-up period all patients remained severely GH deficient, two (12.5%) had partial ACTH insufficiency, one (6.3%) had secondary hypothyroidism but none were gonadotropin deficient or hyperprolactinaemic. CONCLUSIONS: Unlike the severe, evolving multiple pituitary deficits after treatment of pituitary or central tumours in adults, these findings in children with posterior fossa tumours suggest that, with the exception of GH, neurotoxicity due to irradiation per se is associated with a low prevalence of anterior pituitary hormone deficiencies, even at a long follow-up. Since the children in this study were selected for assessment on the basis of growth failure, the high prevalence of GH insufficiency at first testing is to be expected; however, the early onset (within 1-3 years of irradiation) and permanence we have identified supports the view that GH is the most sensitive hormone to radiation injury. PMID- 12555250 TI - Strength and functional mobility in children with acute lymphoblastic leukemia. AB - BACKGROUND: The purposes of this pilot study were as follows: (1) to evaluate tools for measuring neuropathy in children with acute lymphoblastic leukemia (ALL), (2) to examine strength and functional mobility in children with ALL, and (3) to determine if there is a relationship between strength and function. PROCEDURE: Strength of knee extension and ankle dorsiflexion as measured with a dynamometer and functional mobility with the Timed Up and Go (TUG) were examined in eight children with ALL, ages 4-15 years, before and during delayed intensification (DI) therapy and in age- and gender-matched controls. RESULTS: The mean knee extension strength, ankle dorsiflexion strength, and TUG measures for children with ALL before DI were significantly less than the means for the controls. The mean dorsiflexion strength measures for the children with ALL after 4 weeks of DI therapy were significantly lower than at time zero. Correlation of 0.794 (P = 0.05) was found between knee extension strength and TUG score for children with ALL. CONCLUSIONS: The dynamometer and TUG are reliable tools to measure strength and function in children with ALL early in their treatment. In this study, ankle dorsiflexion strength worsened during DI therapy. There was a correlation between strength and function in children with ALL. PMID- 12555251 TI - Characteristics and adequacy of intravenous morphine infusions in children in a paediatric oncology setting. AB - BACKGROUND: Pain management is central in the care of patients in paediatric oncology. Intravenous (i.v.) morphine infusion is an important treatment modality. The objectives of this study were to identify the characteristics of children who receive i.v. morphine infusions in a paediatric oncology setting and to describe the effectiveness, tolerability, and limiting side-effects of this treatment. PROCEDURE: We prospectively collected evaluation charts for all morphine infusions in a paediatric oncology unit during a 4-year period, and reviewed the medical records of the patients. The incidence of breakthrough pain was used as a measure of the effectiveness of morphine treatment. For comparison, patients were grouped according to causes of pain; minor surgery, major surgery, tumour/disease, or treatment side-effects. RESULTS: Overall, 72 individuals (36% of all 201 patients diagnosed in the unit) were given morphine infusions. Patients with pain after major surgery suffered from more breakthrough pain than the other patients during the 1st day of morphine treatment even though their morphine consumption was almost twice as high (0.64 vs. 0.36 mg/kg, P < 0.001); 92% of these patients had solid tumours. Common morphine side-effects were vomiting (38%), nausea (32%), and constipation (24%). CONCLUSIONS: Patients who undergo major tumour surgery require extensive pain management to achieve acceptable post-operative pain relief. The routine of giving both a bolus dose and increasing the morphine dose at breakthrough pain should be encouraged. For the other patients studied, morphine infusions were considered to be adequate, but vomiting, nausea, and constipation might be reduced if prophylactic antiemetics and laxatives were given. PMID- 12555252 TI - Degree of fatness after treatment of malignant lymphoma in childhood. AB - BACKGROUND: Sex hormone deficiency or growth hormone deficiency may cause excess fatness after treatment for childhood malignant lymphoma. Previous studies of the body composition after treatment for childhood cancer included few survivors of malignant lymphoma who were not analysed separately. PROCEDURE: We measured the whole-body percent fat by dual energy X-ray absorptiometry (DXA) and the body mass index (weight/height(2) (kg/m(2)), BMI) in survivors of childhood Hodgkin disease (n = 23) or non-Hodgkin lymphoma (n = 21) a median of 11 years after diagnosis (range 2-25). Results were compared with local data on 463 healthy controls. RESULTS: Adjusted for sex and age, the mean BMI did not differ from that of local controls, but the mean whole-body percent fat was significantly increased (0.8 SD above predicted, P = 0.0001). Sixteen of 44 participants had a percent fat above the 90 percentile of the reference values, which indicates excess fatness. Adjusted for sex and age, percent fat was significantly higher in persons treated for non-Hodgkin lymphoma. Controlled for this, the whole-body percent fat was not significantly related to sex, age at diagnosis, length of follow-up, sex hormone therapy at follow-up or the cumulative dose of corticosteroids or doxorubicin. CONCLUSIONS: Eleven years after diagnosis of childhood Hodgkin disease or non-Hodgkin lymphoma, the whole-body percent fat was increased whereas the BMI was like that of the controls. This indicates a reduced lean body mass. PMID- 12555254 TI - Fatal hemophagocytic syndrome as initial manifestation of a mediastinal germ cell tumor. PMID- 12555253 TI - Valid informed consent and participative decision-making in children with cancer and their parents: a report of the SIOP Working Committee on psychosocial issues in pediatric oncology. AB - This is the tenth official document of the SIOP Working Committee on psychosocial issues in pediatric oncology, instituted in 1991. It is addressed to the pediatric oncology community. Children clearly have a right to participate in medical decisions regarding their own treatment, based on the developmental level of the child. The objective of these guidelines is to encourage physicians to share with the child developmentally relevant medical information specific to that particular child's health status, in the context of the child's own culture, so that he or she can actively participate in the decision-making process regarding his or her own health. These guidelines, geared toward this objective, discuss the child's right to medical information, the parents' legal responsibility for but not exclusive right over the child's health, and ways of encouraging the younger child's active participation in his or her own health care at an age-appropriate level of understanding. For adolescents, there should be a full and legally mandated power to make their own decisions regarding medical treatment. PMID- 12555255 TI - Home platelet transfusion in pediatric oncology terminal care. PMID- 12555256 TI - Disseminated nasopharyngeal natural killer cell lymphoblastic lymphoma in a child. PMID- 12555257 TI - Immunosuppressive treatment for chemotherapy-resistant multisystem Langerhans cell histiocytosis. PMID- 12555258 TI - Is the Epstein Barr virus implicated in Ewing sarcoma? PMID- 12555259 TI - Diabetes insipidus in a child with a monosomy-7 associated myelodysplastic syndrome and neurofibromatosis I. PMID- 12555260 TI - Brain stem glioma in a child: false diagnosis of radiation necrosis with FDG PET. PMID- 12555261 TI - Emily Blackwell (1826-1910) MD: the other Blackwell physician. PMID- 12555262 TI - Medulloblastoma with extensive nodularity in nevoid basal cell carcinoma syndrome. PMID- 12555263 TI - "Two hits" in utero? PMID- 12555264 TI - Chemotherapy alone for the treatment of congenital fibrosarcoma: is surgery always needed? PMID- 12555265 TI - Survey of immunisation practices in the United Kingdom during and following completion of anti-cancer chemotherapy in children. PMID- 12555266 TI - Successful desensitization to trimethoprim-sulfamethoxazole after allogeneic haematopoietic stem cell transplantation: preliminary observations. PMID- 12555267 TI - Vitamin A deficiency leads to increased cell proliferation in olfactory epithelium of mature rats. AB - We have shown previously that vitamin A deficiency (VAD) leads to the decreased expression of gene products that are specifically synthesized by mature neurons in the olfactory epithelium (OE) of adult rats. These results support the hypothesis that retinoic acid, a derivative of vitamin A, is required for neurogenesis and neuron replacement in vivo. VAD does not cause gross degeneration of the OE, raising the question: what types of cells continue to populate VAD OE? In this study, we compared the cell densities of VAD and VA sufficient (VAS) OE and investigated whether cell proliferation is upregulated in VAD OE. The results show that (1) total cell number in VAD and VAS OE are comparable; (2) localized areas of hyperplasia are present in the basal regions of VAD, but not VAS, OE; (3) there is a substantial increase in the number of PCNA (proliferating cell nuclear antigen) positive cells in the basal region of VAD OE relative to VAS OE; and (4) there is a relative increase in the levels of mRNA encoding the transcription factor, MASH I, in VAD OE. We conclude that reduced availability of vitamin A derivatives, such as retinoic acid, leads to a loss of control over proliferation, hyperplasia, and increased numbers of pro neural cells in vivo. PMID- 12555268 TI - Initial stages of neural regeneration in Helisoma trivolvis are dependent upon PLA2 activity. AB - Neuronal regeneration after damage to an axon tract requires the rapid sealing of the injured plasma membrane and the subsequent formation of growth cones that can lead regenerating processes to their appropriate target. Membrane sealing and growth cone formation are Ca(2+)-dependent processes, but the signaling pathways activated by Ca(2+) to bring about these effects remain poorly understood. An in vitro injury model was employed in which neurites from identified snail neurons (Helisoma trivolvis) were transected with a glass microknife, and the formation of new growth cones from the distal portions of transected neurites was recorded at defined times after transection. This study presents three main results. First, phospholipase A(2) (PLA(2)), a calcium-activated enzyme, is necessary for membrane sealing in vitro. Second, PLA(2) activity is also required for the formation of a new growth cone after the membrane has sealed successfully. Thus, PLA(2) plays a dual role by affecting both growth cone formation and membrane sealing. Third, the injury-induced activation of PLA(2) by Ca(2+) controls growth cone formation through the production of leukotrienes, secondary metabolites of PLA(2) activity. Taken together, these results suggest that the injury-induced Ca(2+) influx acts via PLA(2) and leukotriene production to assure growth cone formation. These findings indicate that events that cause an inhibition of PLA(2) or lipoxygenases, enzymes that produce leukotrienes, could result in the inability of neurites to regenerate. PMID- 12555269 TI - Activity alters muscle reinnervation and terminal sprouting by reducing the number of Schwann cell pathways that grow to link synaptic sites. AB - In partially denervated rodent muscle, terminal Schwann cells (TSCs) located at denervated end plates grow processes, some of which contact neighboring innervated end plates. Those processes that contact neighboring synapses (termed "bridges") appear to initiate nerve terminal sprouting and to guide the growth of the sprouts so that they reach and reinnervate denervated end plates. Studies conducted prior to knowledge of this potential involvement of Schwann cells showed that direct muscle stimulation inhibits terminal sprouting following partial denervation (Brown and Holland, 1979). We have investigated the possibility this inhibition results from an alteration in the growth of TSC processes. We find that stimulation of partially denervated rat soleus muscle does not alter the length or number of TSC processes but does reduce the number of TSC bridges. Stimulation also reduces the number of TSC bridges that form between end plates during reinnervation of a completely denervated muscle. The nerve processes ("escaped fibers") that normally grow onto TSC processes during reinnervation are also reduced in length. Therefore, stimulation alters at least two responses to denervation in muscles: (1) the ability of TSC processes to form or maintain bridges with innervated synaptic sites, and (2) the growth of axons along processes extended by TSCs. PMID- 12555270 TI - Olfactory neurons expressing identified receptor genes project to subsets of glomeruli within the antennal lobe of Drosophila melanogaster. AB - We have used green fluorescent protein to trace the projection patterns of olfactory neurons expressing identified candidate odorant receptors to the brain of Drosophila. At the periphery, receptor expression correlates with specific sense-organ subtype, independent of location on the antennal surface. The majority of neurons expressing a given receptor converge onto one or two major glomeruli as described previously. However, we detected a few additional glomeruli, which are less intensely innervated and also tend to be somewhat variable. This means that functionally similar olfactory neurons connect to small subsets of glomeruli rather than to a single glomerulus as believed previously. This finding has important implications for our understanding of odor coding and the generation of olfactory behavior. PMID- 12555271 TI - Seasonal regulation of NMDA receptor NR2B mRNA in the adult canary song system. AB - Developmental changes in the composition and function of N-methyl-D-aspartate receptors (NMDARs) are believed to regulate neural plasticity. For example, in songbirds, vocal learning entails NMDAR activation, and the sensitive period for such learning in zebra finches (ZFs) parallels developmental changes in NMDAR density and phenotype within several song-related brain regions. In contrast to ZFs, canaries exhibit vocal plasticity recurrently throughout adulthood, prompted by seasonal changes in day length and testosterone (T) levels. We used in situ hybridization to determine if such changes in photoperiod affect NMDAR subunit expression in adult canaries. Birds were sacrificed while on short days (SD) when T levels were low, or on long days (LD) when T levels were high. Transcript levels for the constitutive NMDAR subunit (NR1) and two modulatory subunits (NR2A, NR2B) were measured in four song control nuclei: lMAN, Area X, HVc, and RA. NR1 and NR2A mRNA levels were comparable in SD and LD groups in all four song regions studied. However, NR2B mRNA levels within lMAN and RA were significantly higher in SD than in LD birds. Photoperiod did not affect NR2B transcript levels in Area X, HVc, or a nonsong region just lateral to lMAN. Our data support the hypothesis that changes in NMDAR subunit expression may contribute to the neural and behavioral reorganization that accompanies seasonal song remodeling in adulthood. PMID- 12555272 TI - Convergence of mechanosensory inputs onto neuromodulatory serotonergic neurons in the leech. AB - By the frequency-dependent release of serotonin, Retzius neurons in the leech modulate diverse behavioral responses of the animal. However, little is known about how their firing pattern is produced. Here we have analyzed the effects of mechanical stimulation of the skin and intracellular stimulation of mechanosensory neurons on the electrical activity of Retzius neurons. We recorded the electrical activity of neurons in ganglia attached to their corresponding skin segment by segmental nerve roots, or in isolated ganglia. Mechanosensory stimulation of the skin induced excitatory synaptic potentials (EPSPs) and action potentials in both Retzius neurons in a ganglion. The frequency and duration of responses depended on the strength and duration of the skin stimulation. Retzius cells responded after T and P cells, but before N cells, and their sustained responses correlated with the activity of P cells. Trains of five impulses at 10 Hz in every individual T, P, or N cell in isolated ganglia produced EPSPs and action potentials in Retzius neurons. Responses to T cell stimulation appeared after the first impulse. In contrast, the responses to P or N cell stimulation appeared after two or more presynaptic impulses and facilitated afterward. The polysynaptic nature of all the synaptic inputs was shown by blocking them with a high calcium/magnesium external solution. The rise time distribution of EPSPs produced by the different mechanosensory neurons suggested that several interneurons participate in this pathway. Our results suggest that sensory stimulation provides a mechanism for regulating serotonin-mediated modulation in the leech. PMID- 12555273 TI - Targeted gene expression in Drosophila dopaminergic cells using regulatory sequences from tyrosine hydroxylase. AB - Dopamine (DA) is the only catecholaminergic neurotransmitter in the fruit fly Drosophila melanogaster. Dopaminergic neurons have been identified in the larval and adult central nervous system (CNS) in Drosophila and other insects, but no specific genetic tool was available to study their development, function, and degeneration in vivo. In Drosophila as in vertebrates, the rate-limiting step in DA biosynthesis is catalyzed by the enzyme tyrosine hydroxylase (TH). The Drosophila TH gene (DTH) is specifically expressed in all dopaminergic cells and the corresponding mutant, pale (ple), is embryonic lethal. We have performed ple rescue experiments with modified DTH transgenes. Our results indicate that partially redundant regulatory elements located in DTH introns are required for proper expression of this gene in the CNS. Based on this study, we generated a GAL4 driver transgene, TH-GAL4, containing regulatory sequences from the DTH 5' flanking and downstream coding regions. TH-GAL4 specifically expresses in dopaminergic cells in embryos, larval CNS, and adult brain when introduced into the Drosophila genome. As a first application of this driver, we observed that in vivo inhibition of DA release induces a striking hyperexcitability behavior in adult flies. We propose that TH-GAL4 will be useful for studies of the role of DA in behavior and disease models in Drosophila. PMID- 12555274 TI - Wasp venom blocks central cholinergic synapses to induce transient paralysis in cockroach prey. AB - The parasitoid wasp Ampulex compressa induces a set of unique behavioral effects upon stinging its prey, the cockroach. It stings into the first thoracic segment inducing 2 to 3 min of transient flaccid paralysis of the front legs. This facilitates a second sting in the cockroach's head that induces 30 min of excessive grooming followed by a 2 to 5-week long lethargic state. In the present study, we examine the immediate effect of the first sting, which is a transient paralysis of the front legs. Using radiolabeled wasps, we demonstrate that the wasp injects its venom directly into the cockroach's first thoracic ganglion. The artificial injection of milked venom into a thoracic ganglion abolishes spontaneous and evoked responses of the motoneurons associated with leg movements. To investigate the physiological mechanism of action of the venom, we injected venom into the last abdominal ganglion of the cockroach, which houses a well-characterized cholinergic synapse. Injected venom abolishes both sensory evoked and agonist-evoked postsynaptic potentials recorded in the postsynaptic neuron for 2 to 3 min without affecting action potential propagation. Thus, the venom blocking effect has a postsynaptic component that follows the same time course as the transient paralysis induced by the thoracic sting. Finally, injection of a nicotinic antagonist in the front thoracic ganglion induces paralysis of the front legs. We conclude that the transient paralytic effect of the thoracic sting can be mainly accounted for by the presence of a venom active component that induces a postsynaptic block of central cholinergic synaptic transmission. PMID- 12555275 TI - Estrogen regulates tyrosine hydroxylase expression in the neonate mouse midbrain. AB - Estrogen plays an important role during differentiation of midbrain dopaminergic neurons. This is indicated by the presence of estrogen receptors and the transient expression of the estrogen-forming enzyme aromatase within the dopaminergic cell groups. We have previously shown that estrogen regulates the plasticity of dopamine cells through the stimulation of neurite growth/arborization. In this study, we have analyzed the capability of estrogen to influence the activity of developing mouse dopamine neurons. The expression of tyrosine hydroxylase (TH) was assessed by competitive RT-PCR and Western blotting. The developmental expression of TH in the ventral midbrain was studied from embryonic day 15 until postnatal day 15 and revealed highest TH levels early postnatally. This profile coincides with the transient aromatase expression in this brain area. Using cultured midbrain cells, we found that estrogen increased TH mRNA/protein levels. The application of the estrogen receptor antagonist ICI 182,780 resulted in a complete inhibition of estrogen effects. To verify these data in vivo, fetuses were exposed in utero from E15 until birth to the aromatase inhibitor CGS 16949A or to CGS supplemented with estrogen. CGS caused a robust reduction in TH mRNA/protein levels in the midbrain, which could be restored by estrogen substitution. Taken together, our data strongly suggest that estrogen controls dopamine synthesis in the developing nigrostriatal dopaminergic system and support the concept that estrogen is implicated in the regulation of ontogenetic steps but also in the function of midbrain dopamine neurons. PMID- 12555276 TI - Analgesia induced by local plantar injections of opiates in the formalin test in infant rats. AB - Morphine injected locally to the paw of an adult or an infant rat is analgesic. Opiates specific to micro and kappa opioid receptors, and less consistently to delta opioid receptors, given locally to the site of injury in adult animals are also analgesic in a variety of models of inflammatory pain. To determine which opioid receptor(s) are involved in local analgesia in the immature animal, agonists specific for micro, kappa, and delta opioid receptors were injected into the intraplantar pad in infant rats and the resultant nociceptive behavior and Fos expression assayed in the formalin test. The kappa opioid receptor agonist U50,488 reduced nociceptive behavior in both phases of the formalin test and reduced Fos expression in the dorsal horn of the lumbar spinal cord, at 3 and 21 days of age. Morphiceptin (micro opioid agonist) was analgesic in the 21-day-old pups, but not the 3-day-old pups, measured behaviorally or by Fos expression. DPDPE (delta opioid agonist) was not analgesic at either age. We also tested the effects of opioid receptor antagonists on morphine's local analgesic action. Naltrexone, and to a lesser extent the micro opioid antagonist CTOP, antagonized morphine's analgesic effect. Kappa and delta opioid receptor blockers were inactive. The results demonstrate the ability of the kappa opioid system to mediate analgesia in the neonate at the site of injury in acute and chronic pain models, that the micro opioid agonists are active later in development, but that morphine is analgesic in part through micro opioid receptors. PMID- 12555277 TI - Ontogenetic differences in the expression of conditioned visual aversions. AB - We examined ontogenetic differences in the expression of conditioned visual aversions. Sprague-Dawley-derived rats, 16 or 21 days of age, were conditioned with either an element (brightness) or compound (brightness/odor) CS+ and tested for their aversion to the common element (brightness). Aversions to the brightness cue were assessed by either a traditional test of preference between the CS+ brightness and a contrasting brightness or by assessment of freezing in the presence of either brightness cue. The results indicated that strength of conditioning as well as the expression of overshadowing/potentiation was dependent on the age of the animal and on the technique used to assess conditioning. PMID- 12555278 TI - Responses of weaned lambs to fear-eliciting situations: origin of individual differences. AB - The aims of this study were 1) to develop methods of objectively measuring fear in weaned lambs, and 2) to evaluate the effects of age, sex, breed, and rearing conditions on fear reactions. Four hundred forty-one lambs, aged 3 to 6 months, were submitted to three fear-eliciting situations (isolation, surprise effect, and human presence). Factor analysis revealed a first factor, interpreted in terms of fear, that accounted for 40% of total variance in the three tests. As in adult sheep, the main signs of fear were inhibition of feeding, long distance from the frightening stimulus, frequent immobilizations, and numerous high-pitch bleats. Behavior was also influenced by differences in general locomotor activity and exploratory motivation. Fear reactions were influenced by age (3- to 4-month old lambs more fearful than 5- to 6-month-old), sex (females more fearful than males), breed (Romanov more fearful than Ile-de-France), and rearing conditions (artificial vs. maternal: almost no influence in males and influence in females depending on age). These results with sheep provide interesting theoretical and practical perspectives to the study of fearfulness. PMID- 12555279 TI - Age-related differences in neural correlates of face recognition during the toddler and preschool years. AB - Research on the development of face recognition in infancy has shown that infants respond to faces as if they are special and recognize familiar faces early in development. Infants also show recognition and differential attachment to familiar people very early in development. We tested the hypothesis that infants' responses to familiar and unfamiliar faces differ at different ages. Specifically, we present data showing age-related changes in infants' brain responses to mother's face versus a stranger's face in children between 18 and 54 months of age. We propose that these changes are based on age-related differences in the perceived salience of the face of the primary caregiver versus strangers. PMID- 12555280 TI - Effects of prenatal cocaine exposure and maternal separation on heart rate, orienting response habituation, and retention. AB - Effects of the presence or absence of the dam during testing and the retention interval on pretone heart rate (HR) and habituation and retention of an HR orienting response to tone were examined in prenatally cocaine-exposed and nontreated Sprague Dawley rat pups in two experiments. On postnatal day 16, each pup received two test sessions, separated by a 4-hr retention interval during which pups were either isolated or placed with their dam and siblings. For testing, each pup was placed in the test apparatus in the presence or absence of an anesthetized dam where, after a 15-min adaptation period, 10 tone presentations were given, each separated by a 65-s intertrial interval, with HR measured during a 5-s pretone period and throughout the 10-s tone for each trial. Experiment 1 used offspring from the regular breeding colony and observed the typical HR lowering effect of maternal presence during testing, an effect that was surprisingly potentiated, however, following the retention interval in animals that were isolated during this interval. This apparent potentiation by prior isolation of the HR lowering effect of the dam was confirmed in Experiment 2 in nontreated offspring, but did not emerge convincingly in offspring of either dams subcutaneously injected with 40 mg/kg of cocaine HCl daily from gestational days 8 to 20 (C40) or dams injected with saline and pair-fed 4 days (PF4) to mimic the acute anorexic effects of cocaine administration. Consistent with prior work, C40 offspring also were found to exhibit better retention of the habituated orienting response than offspring of NT dams and to some extent PF4 dams as well, a retention effect that was not significantly influenced, however, by social context during the retention interval. PMID- 12555281 TI - Olfactory experience mediates response to pain in preterm newborns. AB - We assessed the effects of a familiar odor during routine blood draws in healthy preterm newborns. Infants were observed as they were undergoing either a capillary puncture on the heel (heelstick) or a venous puncture on the hand. During the procedure, one third of the infants were presented with an odor they had been familiarized with prior to the procedure, one third of the infants were presented with an odor, they had not been previously exposed to, and one third were presented with no odor. Heelsticks elicited more behavioral distress than venipunctures. Infants who were presented with a familiar odor during venipuncture showed no significant increase in crying and grimacing during the procedure compared to baseline levels. By comparison, infants presented with an unfamiliar odor or with no odor either during the heelstick or the venipuncture had a significant increase in crying and grimacing. When the pain was milder, i.e., during a venipuncture, and a familiar odor was presented, infants showed little to no crying. These results are consistent with a body of evidence on early memory and olfactory competence in fetuses and newborns. PMID- 12555282 TI - Classical conditioning in the rat fetus. III. Retention, extinction, and re activation of the conditioned response (CR). AB - A single paired presentation of an artificial nipple (conditioned stimulus, CS) and milk (unconditioned stimulus, US) resulted in classical conditioning. When re exposed to the artificial nipple CS after conditioning, fetuses showed fewer oral grasp responses compared to control fetuses exposed to the milk (US) and artificial nipple (CS) in an unpaired fashion. The reduction in oral grasping was evident when the test of oral grasping was administered 18 min after conditioning but not 21 to 30 min after conditioning (Experiments 1a and 1b). Experiments 2a and 2b confirmed that nine or more exposures to the CS after conditioning, without a milk infusion, resulted in experimental extinction of the conditioned reduction in oral grasping. One re-exposure to milk during the period after classical conditioning was sufficient to re-activate memory for the association between artificial nipple CS and milk US (Experiment 3a). Fetuses exposed to one or two re-activation treatments that comprised an intraoral infusion of milk showed evidence of the conditioned reduction in oral grasping when tested after a delay sufficient to produce forgetting. Experiment 3b showed that the orosensory properties of milk were sufficient to re-activate fetal memory for the association between artificial nipple and milk. Experiment 3c indicated that central injection of 10 ng of dynorphin A 1-13 re-activated fetal memory for the classically conditioned association. Experiments 3b and 3c suggested that the orosensory properties of milk activated a kappa opioid system in the fetal brain that reduced oral grasping of the artificial nipple during the test for oral grasping. PMID- 12555284 TI - Social, thermal, and temporal influences on isolation-induced and maternally potentiated ultrasonic vocalizations of rat pups. AB - Sensory and temporal factors have been demonstrated to be involved in the regulation of isolation-induced ultrasonic vocalizations (USV) of young rats. Sensory cues include thermal, olfactory, and tactile modalities. Temporal factors include the time spent in isolation. The goal of the present research was to examine the interaction of these factors in both isolation-induced and maternally potentiated USV. Maternal potentiation of USV occurs when a brief interaction with the dam, even a passive (anesthetized) dam, elicits an augmented vocal response to a subsequent isolation, with rates of USV in rat pups well above those emitted in standard isolation tests. We found that passive maternal potentiation of USV did occur under all conditions tested. Neither a 30-min prior isolation nor high ambient temperature prevented an increase in USV rate over the rate of the original isolation. After 30-min isolation at warm temperatures when the rate of USV had fallen to zero, the pups increased vocalization in the presence of the dam as well as in the subsequent isolation. Temporal and thermal factors also interacted significantly in regulating the level of the USV emitted by the pups during the first isolation, in the presence of the anesthetized dam, and during the second isolation. PMID- 12555283 TI - Maternal effects in infant and adult phenotypes of 5HT1A and 5HT1B receptor knockout mice. AB - The influence of the pre- and postweaning maternal environment on the offspring's phenotype was examined in 5-HT1A and 5-HT1B receptor knockout mice (KO1A and KO1B, respectively). We have previously shown that, when born to and raised by homozygous dams of the same genotype, adult KO1A are more anxious than wild-type (WT) mice, and adult KO1B are hyperactive and slightly less anxious than WT mice. We extend our studies here to the behavioral results of the offspring's own genotype, when the dam's genotype is constant, and the effects of the dam's genotype when the offspring's genotype is constant. In Experiments 1 and 2, KO1A /- pups produced less ultrasonic vocalizations (USV) than controls in an isolation test on postnatal Day 7 when born to and reared by KO1A dams, either -/ or +/-. Heterozygous F1 pups reared by KO1A-/- dams produced more USV and were less anxious in the plus-maze at 2 to 3 months of age than F1 pups born to and reared by WT dams (Experiment 3). F1 pups reared by KO1B-/- dams produced less USV and were more anxious in the plus-maze than F1 pups reared by WT dams (Experiment 4). The results support a role for maternal effects that may comprise direct effects such as the dam's behavior and nutritional care of the pup, and possibly more complex indirect effects through the establishment of idiosyncratic dam-pup dyadic interactions. We recommend that breeding techniques that rely on same genotype (mutant-mutant or WT-WT) breeding pairs not be used to generate offspring when the focus of research is the study of gene function, but rather when familial effects need to be studied. PMID- 12555286 TI - Significance of apoptosis in surgical oncology. PMID- 12555287 TI - Best practice in groin hernia repair. PMID- 12555288 TI - Growth factors in the treatment of diabetic foot ulcers. AB - BACKGROUND: Chronic foot ulceration is a major source of morbidity in diabetic patients. Despite traditional comprehensive wound management, including vascular reconstruction, there remains a cohort of patients with non-responding wounds, often resulting in amputation. These wounds may benefit from molecular manipulation of growth factors to enhance the microcirculation. METHODS: A review of the current literature was performed using Pubmed, with secondary references obtained from key articles. RESULTS AND CONCLUSION: There has been a generally disappointing clinical outcome from growth factor trials, although topical platelet-derived growth factor has shown significant benefit and should be considered in non-healing, well perfused ulcers after failure of conventional wound care. The modulatory role of the extracellular matrix in the cellular response to growth factors and data from regenerative-type fetal wound healing are further areas of interest. The chemical induction of microvessel formation may become a future therapeutic option. PMID- 12555289 TI - Septic complications after treatment of haemorrhoids. AB - BACKGROUND: Recent reports of serious sepsis following stapled haemorrhoidectomy have raised concerns about the appropriate treatment of haemorrhoidal disease. METHODS: A Medline search was undertaken for reports of sepsis following the commonly practised conservative and surgical treatments of haemorrhoids. RESULTS: Published accounts of significant septic complications after injection sclerotherapy, rubber-band ligation, cryotherapy, open and closed haemorrhoidectomy, and stapled haemorrhoidectomy are discussed. This is supplemented by the authors' own experiences of stapled haemorrhoidectomy. CONCLUSION: Septic complications following both conservative and surgical treatment of haemorrhoids are rare but may be catastrophic. Immunological compromise poses an additional risk for many treatment modalities. The technique of stapled haemorrhoidectomy should be learned diligently to avoid septic complications. PMID- 12555290 TI - The Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity (POSSUM). AB - BACKGROUND AND METHODS: The development of the Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity (POSSUM) is described and its methods of analysis and value in a modern surgical practice are reviewed. A computerized search of all published data in Medline, the Cochrane Library and Embase was made for the last 12 years. Relevant articles were then searched manually for further papers on risk analysis, case-mix comparison and POSSUM methodology. RESULTS AND CONCLUSION: POSSUM has been evaluated extensively in both general and specialist surgery. While there are problems with both data collection and analysis, when used correctly POSSUM can usefully compare outcomes between surgeons and between hospitals. In specialist surgery, individual regression equations may be needed for each index procedure. PMID- 12555291 TI - Control of emboli in patients with recurrent or crescendo transient ischaemic attacks using preoperative transcranial Doppler-directed Dextran therapy. AB - BACKGROUND: Transcranial Doppler (TCD)-directed Dextran 40 treatment after carotid endarterectomy reduces the rate of early postoperative thrombosis. This study assessed the efficacy of intravenous Dextran 40 at controlling symptoms and emboli before elective carotid endarterectomy in patients with recurrent or crescendo transient ischaemic attacks (TIAs). METHODS: In a prospective study, patients with more than 70 per cent internal carotid artery stenosis who had two or more symptomatic episodes within 30 days and TCD-detected microemboli were studied. Dextran 40 was commenced at 20 ml/h and TCD was repeated to reassess the rate of embolization. The infusion was increased in 20-ml/h increments until symptoms and emboli were controlled. The patient then had carotid surgery on the next elective list. RESULTS: Nineteen patients with internal carotid stenosis greater than 70 per cent, recurrent symptoms and TCD-detected emboli were studied. All patients had symptoms and emboli controlled with Dextran 40. One patient with both unstable angina (awaiting urgent operation) and crescendo TIAs died from a myocardial infarct before undergoing operation. Of the 18 patients who had an operation, one suffered a non-disabling stroke on the third postoperative day. CONCLUSION: TCD-directed Dextran 40 offers a safe approach to high-risk patients before elective carotid endarterectomy, and warrants further study. PMID- 12555292 TI - Influence of specialization on the management and outcome of patients with pancreatic cancer. AB - BACKGROUND: Cancer care is increasingly specialized. Relationships between pancreatic cancer care, mortality and patterns of clinical practice among the full spectrum of patients, including those with irresectable tumours, are not well understood. METHODS: A cohort of 782 patients diagnosed prospectively with pancreatic cancer in 29 acute hospitals in England and Wales over 1 year were followed for 2-3 years. The effects of doctors' and hospitals' specialization, indicated by annual patient volumes, on operative mortality rates, survival times, and test and treatment provision were studied. Multiple logistic and Cox regression models were used to control for prognostic factors and treatments, providing adjusted odds and hazard ratios associated with a higher volume of ten patients annually. RESULTS: Patients managed by higher-volume hospitals survived significantly longer (hazard ratio 0.88 (95 per cent confidence interval (c.i.) 0.83 to 0.93); P < 0.001). They were more likely to undergo cytological examination (odds ratio (OR) 1.21 (95 per cent c.i. 1.01 to 1.35)), resection (OR 1.44 (1.17 to 1.79)) and biliary stenting (OR 1.17 (1.02 to 1.34)), and were less likely to have bypass surgery (OR 0.66 (0.55 to 0.78)). Patients of higher-volume doctors were more likely to undergo endoscopic retrograde cholangiopancreatography (OR 1.59 (1.19 to 2.11)), percutaneous transhepatic cholangiography (OR 1.50 (1.12 to 2.00)), laparoscopy (OR 1.81 (1.07 to 3.06)), resection (OR 1.84 (1.29 to 2.61)) and bypass surgery (1.71 (1.25 to 2.33)). CONCLUSION: Specialization appears to improve survival and to promote more thorough investigation. PMID- 12555293 TI - Mapping sentinel nodes in patients with early-stage gastric carcinoma. AB - BACKGROUND: Nodal status in gastric carcinoma is related not only to prognosis but also to the extent of nodal dissection. However, a method for accurate assessment of nodal status during operation has not been established. This study aimed to map the sentinel nodes of gastric carcinoma and to estimate the clinical usefulness of sentinel node biopsy. METHODS: Following laparotomy, a vital dye (0.2 ml 2 per cent patent blue) was injected through a gastroscope into the submucosal layer at four sites around a clinical T1 gastric carcinoma. The dye immediately appeared at the serosal surface and stained lymphatic vessels and nodes. The stained nodes were removed and examined by frozen sectioning. RESULTS: The assay was successful in mapping the lymphatic basins in 203 (96.2 per cent) of 211 patients. The dye stained one or more metastatic nodes in 31 patients, but failed to indicate a metastatic node in four patients with a large involved node. Meticulous postoperative examination of all resected nodes in the standard paraffin slices revealed no new metastases. The accuracy of the assay was 98.0 per cent. CONCLUSION: The method was accurate in predicting nodal status in patients with early-stage gastric carcinoma. PMID- 12555294 TI - Effects of the intermittent Pringle manoeuvre on hepatic gene expression and ultrastructure in a randomized clinical study. AB - BACKGROUND: The intermittent Pringle manoeuvre during hepatectomy results in a better clinical outcome when the accumulated ischaemia time is less than 120 min. The aim of this study was to investigate hepatic gene expression related to microcirculatory modulation and ultrastructural changes in patients having the intermittent Pringle manoeuvre. METHODS: Forty patients who underwent hepatectomy for liver tumours were randomly assigned to liver transection with intermittent Pringle manoeuvre (Pringle group, n = 20) or without the manoeuvre (control group, n = 20). The clinical data and hepatic expression of endothelin (ET) 1 and endothelial nitric oxide synthase (eNOS) combined with liver ultrastructure were compared. RESULTS: The Pringle manoeuvre resulted in less blood loss (8.9 versus 12.4 ml/cm(2); P = 0.034), a shorter transection time (2.7 versus 4.1 min/cm(2); P = 0.015) and a lower serum bilirubin level on postoperative day 2 (26 versus 35 microm/l; P = 0.04). The hepatic messenger RNA content of ET-1 decreased by 38 per cent of the basal level in the Pringle group, whereas it increased by 28 per cent in the control group (P = 0.026). More patients in the control group showed swelling of mitochondria in hepatocytes and disruption of sinusoidal lining cells (12 of 20 patients versus three of 20 in the Pringle group; P = 0.008). CONCLUSION: The intermittent Pringle manoeuvre results in less disturbance of the hepatic microcirculation and better preservation of liver sinusoids after hepatectomy. PMID- 12555295 TI - Pancreatic leak after left pancreatectomy is reduced following main pancreatic duct ligation. AB - BACKGROUND: Although much is known about the long-term outcome of patients undergoing left (distal) pancreatectomy for malignancy, comparatively little is known about the optimal management strategy for the residual transected pancreatic parenchyma and the divided pancreatic duct. Clinicopathological and operative factors that may contribute to postoperative pancreatic leak were evaluated. METHODS: A retrospective review of the medical records of 126 patients who underwent left pancreatectomy between June 1990 and December 1999 at the University of Texas M. D. Anderson Cancer Center was performed. RESULTS: Indications for left pancreatectomy included pancreatic neoplasms (n = 42; 33.3 per cent), en bloc resection for management of retroperitoneal sarcoma (n = 21; 16.7 per cent), gastric adenocarcinoma (n = 14; 11.1 per cent), renal cell carcinoma (n = 11; 8.7 per cent) and other tumours or benign conditions (n = 38; 30.2 per cent). Pancreatic parenchymal closure was accomplished by a hand-sewn technique, mechanical stapling, or a combination of the two in 83, 20 and 15 patients respectively. No form of parenchymal closure was used in eight patients. Identification of the pancreatic duct and suture ligation was performed in 73 patients (57.9 per cent). Twenty-five patients (19.8 per cent) developed a pancreatic leak. For subgroups having duct ligation or no duct ligation, pancreatic leak rates were 9.6 per cent (seven of 73 patients) and 34.0 per cent (18 of 53 patients) respectively (P < 0.001). Multivariate analysis including clinicopathological and operative factors indicated that failure to ligate the pancreatic duct was the only feature associated with an increased risk for pancreatic leak (odds ratio 5.0 (95 per cent confidence interval 2.0 to 10.0); P = 0.001). CONCLUSION: Pancreatic leak remains a common complication after left pancreatectomy. The incidence of leak is reduced significantly when the pancreatic duct is identified and directly ligated during left pancreatectomy. PMID- 12555296 TI - Calcitonin precursors in the prediction of severity of acute pancreatitis on the day of admission. AB - BACKGROUND: Calcitonin precursors are sensitive markers of inflammation and infection. The aim of this study was to evaluate the role of plasma calcitonin precursor levels on the day of admission in the prediction of severity of acute pancreatitis, and to compare this with the Acute Physiology And Chronic Health Evaluation (APACHE) II scoring system. METHODS: Plasma concentrations of calcitonin precursors were determined on admission in 69 patients with acute pancreatitis. APACHE II scores were calculated on admission. Attacks were classified as mild (n = 55) or severe (n = 14) according to the Atlanta criteria. Plasma calcitonin precursor levels were determined with a sensitive radioimmunoassay. RESULTS: On the day of hospital admission, plasma levels of calcitonin precursors were significantly greater in patients with a severe attack compared with levels in those with a mild attack of pancreatitis (median 64 versus 25 fmol/ml; P = 0.014), but the APACHE II scores were no different (median 9 versus 8; P = 0.2). The sensitivity, specificity, positive predictive and negative predictive values, and accuracy for the prediction of severe acute pancreatitis were 67, 89, 57, 93 and 85 per cent respectively for plasma calcitonin precursor levels higher than 48 fmol/ml, and 69, 45, 23, 86 and 50 per cent respectively for an APACHE II score greater than 7. Differences in the specificity and accuracy of the two prognostic indicators were significant (P < 0.001 and P = 0.001 respectively). A plasma calcitonin precursor concentration of more than 160 fmol/ml on admission was highly accurate (94 per cent) in predicting the development of septic complications and death. CONCLUSION: The assay of plasma calcitonin precursors on the day of admission to hospital has the potential to provide a more accurate prediction of the severity of acute pancreatitis than the APACHE II scoring system. PMID- 12555297 TI - Clinicopathological and molecular biological features of colorectal cancer in patients less than 40 years of age. AB - BACKGROUND: The aim of the present study was to identify the clinicopathological and molecular biological characteristics of early-onset colorectal cancers. METHODS: The clinicopathological and molecular biological parameters of 138 consecutive patients with colorectal cancer aged less than 40 years were compared with those of 339 patients aged 60 years or more. RESULTS: The younger patients with colorectal cancer had more mucin-producing (14.5 versus 4.7 per cent; P < 0.001) and poorly differentiated (7.2 versus 3.3 per cent; P = 0.015) tumours, a higher incidence of synchronous (5.8 versus 1.2 per cent; P = 0.007) and metachronous (4.0 versus 0.6 per cent; P = 0.023) colorectal cancers, and more advanced tumour stage (P < 0.001) than older patients. The operative mortality rate was lower (0.7 versus 5.0 per cent; P = 0.026), and cancer-specific survival was similar (in stage I, II and III disease; P > 0.05) or better (in stage IV disease; 95 per cent confidence interval 22.50 to 28.41 versus 12.61 to 17.05 months; P < 0.001). There was a higher percentage of normal p53 expression (61.1 versus 46.8 per cent; P = 0.023) and high-frequency microsatellite instability (MSI-H) (29.4 versus 6.3 per cent; P < 0.001), and a similar family history of cancer (17.5 versus 14.2 per cent; P > 0.05), compared with older patients. CONCLUSION: Young patients with colorectal cancer have several distinct clinicopathological and molecular biological features. The mechanisms underlying the inconsistency between the presence of MSI-H and a family history of cancer in these early-onset colorectal cancers deserve further investigation. PMID- 12555298 TI - Systemic inflammatory response predicts survival following curative resection of colorectal cancer. AB - BACKGROUND: Some studies have shown that the presence of a systemic inflammatory response, as evidenced by raised circulating concentrations of C-reactive protein (CRP), predicted recurrence and overall survival in patients with colorectal cancer. The aim of this study was to examine the relationship between the inflammatory response and overall and cancer-specific survival in patients undergoing potentially curative resection for colorectal cancer. METHODS: A total of 174 patients considered to have undergone curative resection were studied. Circulating CRP concentrations were measured before and/or after operation. RESULTS: The majority of patients were aged 65 years or more, had colonic tumours and Dukes' stage B lesions. During follow-up, 47 patients (27 per cent) developed recurrence and 59 (34 per cent) died. On univariate analysis, age (P < 0.01), Dukes' stage (P < 0.001), and CRP levels before (P < 0.01) and after (P < 0.01) operation were significantly associated with overall and cancer-specific survival. On multivariate analysis of patients in whom preoperative CRP concentration was measured, age (P < 0.01), Dukes' stage (P < 0.05) and CRP concentration (P < 0.01) were independently associated with both overall and cancer-specific survival. CONCLUSION: In patients who have undergone potentially curative resection for colorectal cancer, the presence of a systemic inflammatory response predicts a poor outcome. PMID- 12555299 TI - Treatment of perianal abscess and fistula-in-ano in children. PMID- 12555300 TI - Prospective randomized study of bacteraemia in diathermy and stapled haemorrhoidectomy. AB - BACKGROUND: The incidence and consequences of bacteraemia associated with diathermy and stapled haemorrhoidectomy have not been studied previously. METHODS: Two hundred and five healthy patients randomized to stapled haemorrhoidectomy or diathermy haemorrhoidectomy had perioperative blood cultures taken. The clinical sequelae of bacteraemia and complications of surgery were assessed prospectively. RESULTS: Six patients were excluded for protocol violations. Eleven (11 per cent) of 101 patients with stapled and five (5 per cent) of 98 who had diathermy haemorrhoidectomy had positive blood cultures for organisms after haemorrhoidectomy, predominantly anaerobes commonly found within the bacterial flora of the anorectum (P = 0.19). Transient postoperative pyrexia in several patients did not correlate with detected bacteraemia and settled spontaneously without treatment. There were no serious complications from either operative technique, and no clinical consequences from proven bacteraemia. CONCLUSION: Transient bacteraemia may complicate surgical haemorrhoidectomy but has no serious clinical consequences for healthy adults. PMID- 12555301 TI - Female fecundity before and after operation for familial adenomatous polyposis. AB - BACKGROUND: Knowledge about the fertility of women suffering from familial adenomatous polyposis (FAP) is scarce and inconclusive. The purpose of this study was to investigate the fecundity of women with FAP before and after operation, and to compare the findings with those of a general population database and women with ulcerative colitis. METHODS: A questionnaire concerning reproductive experiences and waiting times to pregnancy was sent to all 230 women on the polyposis registers in Denmark, Finland, Sweden and Norway in whom primary surgery had consisted of ileorectal anastomosis or ileal pouch-anal anastomosis. Data on the general population and women with ulcerative colitis came from an existing database. Cox regression and Kaplan-Meier plots were used for analysis. RESULTS: The fecundity of women with FAP before operation and after colectomy with ileorectal anastomosis was similar to that of the general population. However, fecundity dropped to 54 per cent (P = 0.015) following proctocolectomy with ileal pouch-anal anastomosis, although it was greater than the postoperative fecundity of women with ulcerative colitis. CONCLUSION: The significant reduction in female fecundity after ileal pouch-anal anastomosis should be communicated to young women with FAP before it is decided which surgical option to follow. PMID- 12555302 TI - Factors and consequences of conversion in laparoscopic sigmoidectomy for diverticular disease. AB - BACKGROUND: The disadvantages of laparoscopic elective sigmoidectomy for diverticular disease include the risk of conversion to open operation and longer operative time. The aim of this study was to analyse the causes and consequences of conversion in 168 consecutive patients who underwent a laparoscopically assisted colectomy between January 1994 and June 2001. METHODS: Data were collected prospectively to analyse the causes and consequences of conversion to open surgery in terms of postoperative morbidity and patient recovery. RESULTS: Postoperative mortality, morbidity, conversion and reoperation rates were zero, 21.4 per cent (n = 36), 14.3 per cent (n = 24) and 3.0 per cent (n = 5) respectively. The reasons for conversion were presence of intraperitoneal adhesions and/or inflammatory pseudotumour (n = 21), an intraoperative diagnosis of sigmoid cancer (n = 1), hypercapnia (n = 1) and abdominal bleeding (n = 1). Three preoperative factors were associated with a significant higher risk of conversion: surgical expertise, the presence of sigmoid stenosis or fistula, and the severity of diverticulitis on pathological examination. Morbidity was no different between laparoscopic sigmoidectomy (30 of 144; 20.8 per cent) and converted procedures (six of 24; 25.0 per cent). Open conversion was associated with a longer operative time and significantly delayed patient recovery and hospital discharge. CONCLUSION: Surgical experience and severe diverticular disease are predictive factors for conversion in laparoscopic elective sigmoidectomy. Even if necessary, conversion does not increase the morbidity rate. PMID- 12555303 TI - Extrasphincteric rectal fistulas treated successfully by Soave's procedure despite marked local sepsis. PMID- 12555304 TI - Host immune responses and intestinal permeability in patients with jaundice. AB - BACKGROUND: Systemic endotoxaemia is implicated in the development of complications associated with obstructive jaundice. The aims of these studies were to assess the systemic immune response to intervention in patients with jaundice and to compare the effects of surgical and non-surgical biliary drainage on host immune function and gut barrier function. METHODS: In the first study, 18 jaundiced and 12 control patients were studied to assess systemic immune responses before and after intervention. In the second study, immune responses and gut barrier function were assessed following surgical and non-operative biliary decompression in 45 patients with jaundice. RESULTS: Endotoxin antibody concentrations fell significantly in patients with jaundice immediately after surgical intervention, but not after non-operative biliary drainage. This decrease was associated with a significant increase in serum P(55) soluble tumour necrosis factor (sTNF) receptor concentration (5.3 versus 10.5 ng/ml; P < 0.001), urinary excretion of P(55) TNF receptors (21.4 versus 78.8 ng/ml; P = 0.002) and intestinal permeability (lactulose : mannitol ratio 0.032 versus 0.082; P = 0.048). Intestinal permeability was significantly increased in patients with jaundice compared with controls (0.033 versus 0.015; P = 0.002). CONCLUSION: These data suggest that obstructive jaundice is associated with impaired gut barrier function and activation of host immune function that is exacerbated by intervention. Surgery causes an exaggerated pathophysiological disturbance not seen with non-operative biliary drainage procedures. PMID- 12555305 TI - Fine-needle aspiration cytology in the diagnosis of acute rejection after liver transplantation. PMID- 12555306 TI - Cryotherapeutic ablation of liver tumours (Br J Surg 2002; 89: 1396-1401). PMID- 12555307 TI - Evaluation of axillary lymph nodes using touch imprint cytology and immunohistochemistry (Br J Surg 2002; 89: 1386-1389). PMID- 12555308 TI - Role of magnetic resonance imaging in the diagnosis and single-stage surgical resection of invasive lobular carcinoma of the breast (Br J Surg 2002; 89: 1296 1301). PMID- 12555309 TI - Radio-guided sentinel node detection for gastric cancer (Br J Surg 2002; 89: 604 608). PMID- 12555310 TI - Ranitidine as adjuvant treatment in colorectal cancer (Br J Surg 2002; 89: 1416 1422). PMID- 12555311 TI - Management of popliteal aneurysm (Br J Surg 2002; 89: 1382-1385). PMID- 12555313 TI - Complications and functional outcome following artificial anal sphincter implantation (Br J Surg 2002; 89: 877-881). PMID- 12555315 TI - [Junctional tachycardia in adolescents: nodal reentry is the most frequent cause]. AB - Ventricular preexcitation syndromes are classically more common in the pediatric age group than in adults, and a latent Kent bundle may explain most cases of paroxysmal junctional tachycardia (PJT). These data stem from the results of intracardiac electrophysiologic testing, which is performed only in those patients at the most severe end of the symptom spectrum. The recent introduction of transesophageal testing has expanded the indications of electrophysiologic testing for PJT. This technique was used to determine the mechanism of PJT in 23 adolescents aged 11 to 9 years (mean age, 16 +/- 3 years) with paroxysmal palpitations, accompanied in seven cases with dizziness or syncope. Only four patients had documented PJT. The basal ECG was normal, and exercise testing showed no evidence of preexcitation. Pacing at increasing rates and programmed stimulation with one then two extra-stimuli was used and repeated, if needed, under infusion of 20 to 30 micrograms of isoproterenol. RESULTS: PJT was induced in 21 patients (91%), under basal conditions in 13 and under isoproterenol in eight. Neither of the two patients with a negative test had documented tachycardia. Based on classic criteria (position of A relative to V1, effect of a bundle branch block, and shape of A in D1 and V1), the mechanism of the PJT was shown to be nodal reentry in 17 cases (81%) and reentry into a latent left-sided Kent's bundle in four cases. Atrial fibrillation was also induced in two of the patients with nodal reentry. In six of the patients with dizziness or syncope associated with palpitations, these symptoms were due to nodal reentry. CONCLUSION: Nodal reentry is very common in adolescents and can explain symptoms ascribed to "spasmophilia", as well as some cases of malaise or syncope. PMID- 12555316 TI - [Tolerance to valsartan in office practice in 3,197 hypertensive patients (the VALSE study)]. AB - Valsartan, an angiotensin II antagonist with a potent and highly selective effect on AT1 receptors, has been found effective and safe in several randomized, controlled studies versus a placebo, other antihypertensive agents, or other angiotensin II antagonists. An eight-week, open-label, multicenter, prospective, pragmatic, phase IV trial was conducted by office-based cardiologists to evaluate the clinical safety and efficacy of valsartan 80 mg in everyday practice. The efficacy analysis included 3084 patients given valsartan alone for four weeks then, during the next four weeks, either valsartan alone or, if the diastolic blood pressure was still > or = 90 mmHg, valsartan in combination with another antihypertensive agent. The valsartan dosage was 80 mg/day. On day 28, 64% of patients were controlled and 78% were responders. The safety analysis included 3,108 patients. Of the 239 medical adverse events ascribed by the investigators to the study treatment, 72 led to premature study discontinuation. This figure translated into a 2.68% rate among treated patients, which is similar to the 2.3% rate recorded during the placebo-controlled trials performed as part of the valsartan clinical development program. These results confirm the outstanding safety profile of valsartan. PMID- 12555317 TI - [Post-embolic chronic pulmonary heart disease: a case report]. AB - Echocardiography and pulmonary scintigraphy demonstrated post-embolic chronic cor pulmonale in a six-year-old woman presenting with dyspnoea. The patient's symptoms deteriorated during subsequent thromboembolic episodes, despite treatment consisting of oral anticoagulants and diuretics. After discussing the pathophysiology and diagnostic methods, the authors emphasize the limits of medical treatment of post-embolic chronic cor pulmonale and the value of two surgical treatments: thromboendarterectomy and lung transplantation. They also recall the precise selection criteria for lung transplantation. PMID- 12555318 TI - [End-stage heart failure: role of various circulation assistance techniques]. AB - There are currently 4 main types of cardiac assist devices: centrifugal pumps, paracorporeal pneumatic assist devices, implantable ventricular assist devices, total artificial heart. Although the efficacy of centrifugal pumps remains limited, that of the other 3 types of assist devices has now been clearly demonstrated. The choice between these various devices depends on the severity of heart failure at the time of implantation, the univentricular or biventricular disease, the potential for recovery, the patient's morphology and obviously the device or devices available to the team concerned. The results essentially depend on early implantation. PMID- 12555319 TI - [Etiopathogenesis of arrhythmogenic right ventricular dysplasia]. AB - Arhythmogenic right ventricular dysplasia (ARVD) is a genetically determined cardiomyopathy with a dominant transmission mode and variable penetrance. Transdifferenciation of cardiomyocytes into adipocytes is likely to explain massive replacement of right ventricular and to a lesser extent left ventricular myocardium by adipose tissue. This phenomenon starts in the mediomural layers and extends into the epicardium. It can occur in the fetus, however youth and young adults are more frequently involved. Apoptosis defined as a programmed cell death, is likely to enhance adipogenesis and tiny fibrosis production. Inflammation can be superimposed on the genetically determined substrate and usually involves both ventricles. Myocarditis can be acute or chronic with interstitial or scar fibrosis, or active chronic. In some cases, left ventricular involvement can be as important as right ventricle, characterizing biventricular dysplasia. In Naxos disease, ARVD is associated with an ectodermic dysplasia. The transmission mode is recessive. PMID- 12555320 TI - [Cholesterol crystal embolism]. AB - Cholesterol crystal emboli are a serious complication of atheroma. The incidence of this syndrome appears to be much more common in patients in their sixties with severe atheromatous disease of the aorta (20 to 30% vs less than 5%). 80% of crystal embolism result from medical interventions (aortic or cardiac surgery, arterial invasive procedure of aorta, thrombolytic therapy). Embolisation of cholesterol crystals can give rise to a confusing clinical pictures, depending of the site (organ) of the embolisation. It also can simulate a systemic disease. The diagnosis must be discuss in all atheromatous patients. The prognosis is poor because of the patient's clinical context and because there is no specific treatment. The best treatment is prevention by improving the recognition of the high risk patients and avoiding in those cases the predisposing factors. PMID- 12555321 TI - [Aspirin: towards a new consensus]. PMID- 12555322 TI - [Translation of the recommendations and proceedings of the 5th North American Consensus Conference on Antithrombotic Therapy]. PMID- 12555333 TI - [Value of transesophageal programmed atrial stimulation in the evaluation of unexplained cerebrovascular accidents]. AB - Certain embolic cerebrovascular accidents can be explained by the development of paroxysmal atrial fibrillation. When noninvasive complementary investigations are negative, programmed atrial stimulation can be proposed to detect increased atrial vulnerability. The objective of this study was to evaluate the reliability of this method performed via a transoesophageal approach in 59 subjects presenting with an embolic cerebrovascular accident and who were in sinus rhythm at the time of the accident. Seven of these patients had a history of paroxysmal atrial fibrillation (AF) or atrial tachycardia (AT) (group I). Three of these seven patients also presented AV nodal reentrant junctional tachycardia. The other 52 patients had no history of arrhythmia and their Holter recording did not reveal any episodes of sustained atrial tachycardia (group II). Transoesophageal programmed atrial stimulation used up to 2 extrastimuli under baseline conditions and during Isuprel infusion. The following results were obtained: sustained atrial tachycardia (> 1 min) was induced in all patients of group 1, 3 of them also presented inducible junctional tachycardias. 14 patients of group II (27%) presented inducible supraventricular tachycardia: atrial tachycardia in 7 cases. Patients in group II with inducible AT presented either heart disease (n = 3) or minor abnormalities on the Holter recording (runs of atrial premature complexes or sinus pauses (n = 3). Two of these patients subsequently developed sustained atrial fibrillation during follow-up. In 25 patients with normal Holter recording and no heart disease, programmed atrial stimulation induced junctional tachycardia in 4 cases. In conclusion, transoesophageal electrophysiological investigation is a useful way to identify various forms of supraventricular tachycardia able to explain an embolic cerebrovascular accident. The considerable incidence of inducible AV nodal reentrant junctional tachycardia must be emphasized, while the incidence of atrial fibrillation is much lower than during intracardiac investigations. PMID- 12555334 TI - [CREST syndrome presenting as pulmonary hypertension]. AB - The authors report the case of a 62-year-old patient complaining of recent onset of disabling breathlessness on exertion, and presenting clinical signs of previously undiagnosed scleroderma. Echocardiography revealed a diagnosis of precapillary pulmonary hypertension (74/14 mmHg) (PHT), with no pulmonary cause revealed by pulmonary ventilation-perfusion scintigraphy or by thoracic fine section computed tomography. The diagnosis of PHT in the context of circumscribed scleroderma was confirmed by x-rays of the hands, capillaroscopy, oesophageal investigations and positive anticentromere antinuclear antibodies. The clinical course was marked by rapid deterioration of the symptoms, requiring treatment with prostacyclin by continuous intravenous infusion. The appearance of PHT in a context of circumscribed scleroderma, usually a relatively benign disease, is a rare, late event, exceptionally revealing the disease, as in this case, and indicating a very unfavourable prognosis. PMID- 12555335 TI - [Efficacy of cibenzoline in a case of atrial fibrillation due to cardiac thyrotoxicosis]. AB - The authors report a case of reduction of thyrotoxicosis-related atrial fibrillation by cibenzoline. The initial clinical features of heart failure rapidly resolved and the echocardiographic left ventricular shortening fraction returned to normal. Cibenzoline could be used as antiarrhythmic treatment in atrial fibrillation related to thyrotoxicosis. PMID- 12555336 TI - [Treatment of coronary insufficiency in diabetics: Part 2: acute coronary insufficiency]. AB - Treatment of acute coronary insufficiency in diabetics--recent myocardial infarction (MI--unstable angina--uses the same modalities as in the absence of diabetes. Thrombolytics improve the prognosis of MI, although the hospital mortality remains about two fold in the presence of diabetes. Primary angioplasty has an identical success rate, but restenoses are significantly more frequent in diabetics. Systematic use of stents allows a reduction of the restenosis rate to the level observed in the absence of diabetes. In unstable angina, low molecular weight heparins have an efficacy and a safety identical to those observed in non diabetic patients. There is therefore no limitation to their use. Diabetics present permanent activation of blood platelets which promotes their adhesion and aggregation. Aspirin must therefore be systematically prescribed to diabetic patients, except in the presence of a contraindication, especially gastrointestinal, in which case, ticlopidine can be used. Platelet glycoprotein IIB-IIIA receptor inhibitors have the same indications and provide the same results as in the absence of diabetes. Contrary to a widely held belief, beta blockers, especially cardioselective, can be widely used in diabetics. The same applies to angiotensin converting enzyme inhibitors. Finally, during the acute phase of coronary insufficiency, continuous insulin infusion via a pump ensures better control of diabetes and also decreases the mortality of MI. Permanent collaboration between cardiologists and diabetologists is therefore essential to increase the efficacy of treatment and to improve the prognosis of acute coronary insufficiency in diabetic patients. PMID- 12555337 TI - [Primary and secondary prevention of coronary disease by statins]. AB - In the context of the growing complexity of atherosclerosis, LDL-cholesterol has been gradually revealed to be the main aetiological agent. However, the first clinical trials were not convincing, as they failed to demonstrate a significant improvement of coronary mortality. So-called "regression" trials revealed the need to develop clinical trials in which the primary endpoint is coronary events and no longer intermediate criteria such as the course of coronary disease on coronary angiography. Lovastatin, simvastatin and pravastatin were shown to be effective in the prevention of relapses in coronary patients and in the prevention of coronary events in healthy subjects. In 1999, the prescription for coronary patients should almost always include a statin and the prescription in healthy subjects should include a statin when this subject presents a high risk of coronary events in the years to come. PMID- 12555338 TI - [Evaluation of the efficacy and tolerance of naftidrofuryl in patients presenting with exertional angina. Multicenter double-blind versus placebo study]. AB - Due to its vascular and platelet 5-HT2 receptor antagonist properties and its metabolic properties, naftidrofuryl specifically counteracts local ischaemic phenomena. One of its major indications is the treatment of intermittent claudication, but it is well known that peripheral arterial disease is the sign of diffuse arterial disease, associated with particularly lethal coronary disease. Recent studies increasingly implicate serotonin (5-HT) in coronary ischaemic processes. In view of the similarities between these pathophysiological data and the characteristics of this molecule, we decide to evaluate the coronary protection afforded by naftidrofuryl and its safety. This multicentre double blind placebo-controlled study was conducted in 51 patients over a period of one month. Inclusion criteria were stable angina with an electrically positive stress test, despite antianginal treatment either by beta-blocker or by calcium channel blocker. Follow-up comprised clinical assessment and a stress test on inclusion and at 1 month. The groups were comparable on inclusion. Overall, the results showed a greater improvement with naftidrofuryl than with reference treatment for all parameters studied. Significant differences were observed in favour of the verum group for time to onset of ST depression, the maximum level reached, the number of stress tests which became negative and the patient's global assessment. No problems of interaction with concomitant treatments, particularly beta blockers, calcium channel blockers or antiarrhythmics was observed. This study shows that naftidrofuryl allows improvement of ergometric parameters and especially elevation of the ischaemic threshold on exertion. PMID- 12555339 TI - [Myocardial infarction after a stress test. A case report]. AB - The authors report a case of first myocardial infarction occuring after an asymptomatic stress test in a patient presenting several cardiovascular risk factors. Coronary angiography, performed before the 4th hour after onset of the symptoms in this patient, identified the mechanism of myocardial infarction to be occlusion of the left anterior descending artery by thrombus, very probably complicating rupture of an atherosclerotic plaque. The initial failure of thrombolytic therapy led to a mechanical revascularization procedure with primary success. The pathophysiological mechanisms possibly contributing to this type of accident are discussed. PMID- 12555340 TI - [Right mid-ventricular stenosis with intact interventricular septum]. AB - Pure infundibular stenosis of the right ventricle is a rare congenital anomaly. Right midventricular stenosis with intact interventricular septum is even rarer. It consists of obstruction of the trabecular zone of the right ventricle by a hypertrophied moderator band with or without abnormal muscle bundles. This anomaly can now be diagnosed by transthoracic echocardiography, while haemodynamic and angiographic diagnosis is reserved for doubtful cases. Two cases of midventricular stenosis are reported. In both patients, the diagnosis was suspected bu ultrasonography which showed a right intraventricular gradient of about 80 mmHg without identifying the exact site (infundibular or midventricular). The diagnosis was corrected by the operative findings. A good postoperative course was observed for both patients. PMID- 12555341 TI - [Pericardial mesothelioma. A case report]. AB - Primary pericardial mesothelioma is a rare malignancy, with an estimated incidence of 0.0022% in a large autopsy study. We report a case of primary pericardial mesothelioma revealed by a large and recurrent pericardial effusion. Through a literature review, we analyse the clinical findings of this tumor. With or without therapy, prognosis is poor. PMID- 12555342 TI - [A case of pneumopericardium complicating a liver abscess in a patient infected with HIV]. AB - Pneumopericardium usually occurs in a traumatic context or, more rarely, in the course of benign or malignant bronchial or oesophagogastric diseases. Although the idiopathic spontaneous form is even rarer, pneumopericardium complicating liver abscess is exceptional, and is facilitated by impaired immune defenses (alcoholism, immuno-depression). The authors report a case of liver abscess complicated by pneumopericardium in an HIV seropositive patient. The nature of the abscess, the mechanism of the pneumopericardium and the therapeutic management are discussed. PMID- 12555343 TI - [Cardiac adverse events from non-cardiac drugs]. AB - Prevention of adverse events of non-cardiological medicinal products is essentially based on the knowledge that they exist and quantification of the risk to which the patient is exposed, both in terms of frequency and potential severity. PMID- 12555344 TI - [Ventricular late potentials in myocardial infarction: comparison of thrombolysis, primary angioplasty, and conventional treatment]. AB - OBJECTIVES: The aim of this study was to compare the prevalence of ventricular late potentials (VLP) during the acute phase of myocardial infarction (MI) depending on the treatment used. METHODS AND RESULTS: This retrospective study was carried out over the period January 1992 to December 1997, and involved 238 patients admitted for primary MI and treated less than six hours after the onset of symptoms by intravenous thrombolysis (rt-PA, n = 83) or primary angioplasty (ATCI, n = 82) and in those cases treated over six hours after the onset of symptoms by standard treatment (heparin, n = 73). Rt-PA perfusion was considered to be effective when the three following criteria were simultaneously present: i) pain sedation; ii) regression of the ST dysfunction > 50%; iii) creatine phosphokinase (CPK) peak before the 16th hour. Treatment success for angioplasty (ATCI+) was confirmed by a TIMI 3 (Thrombolysis in Myocardial Infarction) score for MI arterial perfusion, with residual stenosis of the MI artery of < 50%. Ventricular late potentials (VLP) were investigated between day six and 14, and considered to be present when two of the following criteria were met: total duration of filtered QRS, i.e., QRSd > 114 ms; signal amplitude in the 49 last ms of filtered QRS, i.e., RMS40 < 20 mV, duration of the terminal part of filtered QRS that was below 40 mV, i.e., LAS40 > 38 ms (40 Hz). VLP prevalence was as follows: 25% (rt-PA group), 345 (ATCI group), and 33% (standard treatment) respectively (P = NS). In the thrombolysis with reperfusion subgroup (n = 54/83, 65%), VLP incidence was 18.5% (n = 10/54) versus 35.5% (n = 27/76) in the subgroup ACTI+ (n = 76/82, 93%) (P = 0.03). CONCLUSIONS: The prevalence of VLP appears to be significantly decreased in the the case of thrombolysis with reperfusion compared to that observed in the ATCI+ group. One of the possible explanations for this abnormally high prevalence in the angioplasty group could be a dysfunction involving reocclusion after initially successful myocardial reperfusion therapy. PMID- 12555345 TI - [Cardiac lymphoma disclosed by ischemic accident. A case report]. AB - We report on the detection of a primitive cardiac lymphoma revealed by a cerebral vascular accident in a context of deterioration of the general state with fever. The diagnosis of the cerebral vascular accident was verified by the brain scan. The echocardiography, in particular the transesophageal ultrasound, suggested the diagnosis of cardiac lymphoma due to the existence of an inhomogeneous multilobar mass invading the right ventricle and largely encompassing the right auricle. The histologic diagnosis of a malignant non-Hodgkin's lymphoma of type B malignancy was confirmed by anatomopathology. The evolution was, unfortunately, rapidly unfavorable, with the patient dying in a state of cerebral suffering despite a combination treatment of chemotherapy and radiotherapy. PMID- 12555346 TI - [Staphylococcus epidermidis infective endocarditis after mitral surgery, successfully treated with aspirin and antibiotics: a case report]. AB - We report a case of infectious endocarditis from Staphylococcus epidermidis that occurred early after mitral valve repair (one month), suggested by fever of 38.5 degrees C and valvular vegetations of less than 10 mm on the mitral valve. In the absence of standard recommendations (medical or surgical) in treating patients with infectious endocarditis occurring after mitral valve repair, and in the absence of complications, a medical regimen was chosen associating aspirin (anti aggregant dosages) with antibiotics. The benefit of aspirin in endocarditis has been demonstrated in experimental studies with regards to valvular vegetations and embolic risk but remains to be studied in human clinical trials. With the association of aspirin (100 mg/d) and triple antibiotic therapy (rifampicin 1200 mg/d, vancomycin 2 g/d, gentamycin 180 mg/d), the clinical status improved with complete regression of vegetations in less than 24 days and the absence of recurrence at one-year follow-up. PMID- 12555347 TI - [Diastolic function of the left ventricle in a North-African patient with homozygous sickle-cell anemia]. AB - AIM: The aim of this study was to investigate left ventricular diastolic function in a group of homozygous Black African patients with sickle-cell disease via echocardiography-Doppler, and to describe the possible hemodynamic implications of these findings. PATIENTS AND METHODS: Fifty patients with sickle-cell anemia from two hematology departments in Yaounde volunteered to participate in the study, carried out between May-October 1995. The presence of any other associated pathology (cardiovascular disorder, acute drepanocytic anemia, pregnancy) was excluded following a clinical, hematological and echocardiographic examination. A sex- and age-matched control group consisting of 50 healthy subjects was selected after the same examination criteria had been met. Mitral flow recorded by pulsed Doppler was measured, and the amplitudes of the E and A waves of the proto- and telediastolic flow, the E/A ratio, the decrease over time (DT) for the E wave, and the left ventricular isovolumetric relaxation time were also calculated. The limits of the confidence interval for the mean DT of the control group were used as the criterion to determine the following in the patient population: i) relaxation disorder profile, with DT > 154 ms; ii) restrictive disorder profile, with DT < 137 ms. RESULTS: The dimensions of the cardiac cavities and the left ventricular mass were higher in the patient population; 34 cases of left auricular dilatation (68%) and 33 cases of left ventricular dilatation 8 66%) were observed. In only two patients was a systolic dysfunction noted. The amplitudes of the mitral flow E and A waves were increased, and the DT was longer in the sickle-cell group. Forty patients (80%) had abnormal diastolic function, with a marked predominance of the 'relaxation disorder' profile (60%). The restrictive disorder profile was only observed in ten sickle-cell patients (20%), while ten others displayed a normal profile (20%). The means of the various clinical and echocardiographic parameters of the three categories of sickle-cell patients (based on their diastolic profile, i.e., normal, relaxation disorder, restrictive disorder) were compared. The clinical parameters could be superimposed in the 3 categories. The dimensions of the left auricle and the telesystolic diameter of the left ventricle were significantly larger in the restrictive disorder category. The latter category appeared to be more symptomatic, and included the two isolated cases of systolic dysfunction. CONCLUSION: Left ventricular diastolic dysfunction is commonly encountered in sickle-cell cases in the Cameroon. The restrictive disorder profile is less frequently observed, but appears to be more harmful at the hemodynamic level. The present results suggest that the study of diastolic function in patients with sickle-cell anemia could help identify hemodynamically at-risk subjects who require more specific cardiovascular care, even before a noticeable alteration in systolic function has taken place. PMID- 12555348 TI - [Effect of golf on sinus rate variability]. AB - There are no or very few studies in the literature on the effects of golf on heart rate and its parameters. The purpose of the study was to evaluate the effects of a precision sport, golf, on heart rate variability (HRV). The study population consisted of six high-level golfers aged 25 to 40 years, without cardiac disease. HRV was studied three hours before a golf competition, during the four hours of the competition and three hours after the match was over. The following parameters of HRV were calculated during 12 games of golf: mean heart rate (HR), standard deviation of the mean RR intervals (SDNN), root mean square of successive differences in RR intervals among consecutive normal beats (rMSSD), percent differences between normal RR intervals that are greater than 50 ms computed over the entire 24-hour recording (pNN50), low-frequency amplitude (LF), high-frequency amplitude (HF) and LF/HF ratio: ratio of low-to-high frequency power. Analysis of the results indicates significant variations of all parameters which occur just at the beginning of the game, persist throughout the match and remain three hours after its completion: heart rate is increased; total HRV is decreased but parameters reflect parasympathetic activity; pNN50, rMSSD and 1-117 are principally decreased. In conclusion, golf significantly affects HRV just at the beginning of golf competition, probably because of stress. These changes progressively decrease but they are still significant three hours after the end of the competition. PMID- 12555349 TI - [Evaluation of a new preservation solution for hypothermic heart transplant]. AB - AIM: One of the major problems encountered in heart transplant is the limited cardiac preservation time. The time limit from the moment the donor heart is removed to the transplant itself is 4-6 hours at maximum. Extending the preservation time would therefore provide access to a larger pool of donors, and also permit long-distance (transfrontier) organ transfer. To attain this aim, a number of different cardiac preservation solutions have been proposed and evaluated either clinically or experimentally; however, no consensus has yet been reached by the various heart transplant teams involved. As finding an optimal solution is of major importance, the aim of the present study was therefore to assess the efficacy of several cardiac preservation solutions: the University of Wisconsin solutions (UW-1 and UW-1 + calcium = UW-2), the Saint-Thomas' Hospital solution (STH-1) and a new solution (NS) developed by our laboratory. METHOD: Male Wistar rats (n = 60) were anesthetized by i.m. injection of pentobarbitol, and randomized into six groups, i.e., NS (group I), STH-1 (group II), UW-1 (group III), UW-2 (group IV), saline solution, NaCl (group V), and controls (group VI). All the hearts except those in the control group were preserved by immersion (8 hours, 4 degrees C) in the different solutions (n = 10/group). The hearts were either a) reperfused in Langendorff mode (functional assessment, n = 5/group); or b) frozen (energetic assessment, n = 5/group). RESULTS: The hearts treated with NaCl displayed arrhythmia and a collapsed mean left ventricular pressure throughout the reperfusion period (3.6 +/- 1.3 mmHg) compared to the NS hearts and the control group (97 +/- 2.6 and 122 +/- 1.9 respectively), while the STH-1, UW-1 and UW-2 hearts showed an intermediate-range ventricular function (48 +/- 4; 39.9 +/- 4.1; and 69 +/- 1.8 respectively). Creatine kinase levels were higher in the STH-1 and NaCl groups (541.9 +/- 168 and 1,080.8 +/- 126.2 IU/L respectively). The energetic charge was significantly lower in the NaCl and UW-2 groups (P = 0.001) compared to the control, NS, STH-1 and UW-1 groups. CONCLUSION: It was found that the composition of the respective solutions had a major effect on the the quality of myocardial preservation. Compared to the other solutions tested in this study, the NS solution was found to be optimal for the preservation of isolated hypothermic rat hearts. PMID- 12555350 TI - [Harmonic imaging: acquired and congenital cardiopathy]. PMID- 12555351 TI - [Intermittent claudication in a young patient. A case of isolated fibromuscular dysplasia of the external iliac artery]. AB - Effort-linked intermittent claudication of arterial origin in sportsmen is often attributed to endofibrosis of the external iliac artery. Some knowledge of possible differential diagnoses, in particular the fibrodysplasia, is of importance regarding the therapy involved. Angioplasty treatment of external iliac endofibrosis may be controversial. However, the same does not apply to fibrodysplasia angioplasty, particularly if the latter is accomplished by inserting an endoprothesis. A case of fibromuscular dysplasia of external iliac artery in a 37 year old woman, treated with endoluminal angioplasty and stent, is reported. PMID- 12555352 TI - [Horton's disease presenting as a myocardial infarction]. AB - The authors report the case of a 60-year-old man admitted for uncomplicated acute myocardial infarction. The history of persistent fronto-occipital headaches and palpation of bilateral tortuous temporal arteries with a decreased pulse suggested the diagnosis of temporal arteritis, particularly in the presence of marked laboratory signs of inflammation and the coronary angiograph findings. The diagnosis was confirmed by temporal artery biopsy. Treatment consisted of corticosteroid therapy, with a satisfactory outcome at three months. The diagnosis of temporal arteritis should be systematically suggested in patients over the age of 50 years, presenting with a marked inflammatory syndrome and recent-onset arterial disease. Similarly, any form of arterial disease occurring during treated temporal arteritis should initially be considered to be a secondary site of the arteritis, requiring intensification of corticosteroid therapy. PMID- 12555353 TI - [Congenital left ventricular aneurysms and diverticula. Two case reports]. AB - The authors report the cases of two young adults (25 and 27 years) presenting with congenital left ventricular aneurysm or diverticulum with healthy coronary arteries. This saccular evagination of the ventricular wall is rare. The authors describe a classification distinguishing muscular (contractile) diverticula, composed of the three cardiac tunics, fibrous diverticula and finally aneurysms with a dyskinetic wall. Although the limits of this classification are sometimes poorly defined, it presents a prognostic value, because it appears logical to propose nonsurgical management in the context of muscular diverticula and it seems legitimate to operate on fibrous diverticula and aneurysms due to the risks of rupture, extension, thrombosis or arrhythmia. PMID- 12555354 TI - [Treatment of coronary insufficiency in diabetics. Part 1: objectives and targets]. AB - Coronary insufficiency affects 55% of insulin-dependant diabetics and is responsible for 60% of deaths in this population. Its particular severity is essentially due to the severity of coronary atherosclerosis, which is usually multi-vessel, involves both large trunks and microcirculation, is made of frequently lipid-rich and therefore fragile plaques, and is accompanied by abnormal but specific reactions of the arterial wall (tendency to vasoconstriction and increased neointimal proliferation after trauma). Coronary atherosclerosis is also often associated with HT, lower limb arteriopathy or cerebral atherosclerosis. Quality of blood glucose control, other organic lesions of diabetes (nephropathy, retinopathy), disturbances of platelet function and dyslipidaemias (hypercholesterolaemia, hypertriglyceridaemia, increased levels of highly atherogenic small LDL particles) are also involved in the development of coronary insufficiency. A precise knowledge of the diseases to be treated and their particularly rigorous prevention and treatment can improve the prognosis of coronary insufficiency in diabetics. PMID- 12555355 TI - [Endovascular treatment of intracranial aneurysms]. AB - Over recent years, an increasing number of patients with intracranial aneurysms are treated by endovascular techniques, which have the advantage of avoiding craniotomy and retraction of cerebral parenchyma, while reducing the postoperative hospital stay. The key concept is that aneurysm is a disease of the arterial wall and that treatment must ideally only concern the vessel. The short term efficacy of endovascular treatment has been established, especially for the prevention of early rebleeding, but its long-term efficacy has not yet been formally demonstrated due insufficient follow-up. Ideally, the management of intracranial aneurysms should be based on close collaboration between neurosurgeons and interventional neuroradiologists and the approach (conventional surgery or endovascular) should be discussed case by case. PMID- 12555356 TI - [Treatment of refractory cardiac insufficiency by peritoneal ultrafiltration. Literature review]. AB - The management of heart failure that has become refractory to conventional drug treatment is an increasingly frequent problem for clinicians. Peritoneal dialysis, by ultrafiltration, is an effective method to treat refractory oedema, thereby improving the functional symptoms of these patients. The objective of this article is to review the data, of the literature and to define the results and indications of this treatment, which still remains and exceptional modality. PMID- 12555357 TI - [Heparin-induced thrombopenia]. AB - Heparin-induced thrombocytopenia remains a topical subject for at least two reasons. The first reason is the increasing prescription of low molecular weight heparins (LMWH) rather than unfractionated heparins, with limited laboratory surveillance, raising the question concerning the need for twice-weekly platelet counts, according to the recommendations of the Vidal drug directory. The second reason is the recent release onto the market of two products, danaparoid (Orgaran) and lepirudin (Refludin) for this precise indication of heparin-induced thrombocytopenia. These products greatly facilitate the management of this complication. Many basic research teams are trying to optimize the detection of heparin-dependent antibodies and to more clearly elucidate the mechanism of this particular thrombocytopenia, which carries a risk of very severe thrombotic complications when the diagnosis is delayed. PMID- 12555358 TI - [Cardiac surgery in the octogenarian]. AB - Progressive ageing of the population results in an increased number of patients in their eighties referred for cardiac surgery. The combination of the effects of ageing and age-related diseases makes eighty-year-olds a group of high-risk surgical patients. Due to the increasingly limited resources of health care systems, the results of cardiac surgery in eighty-year-olds must be re-evaluated in terms of survival and postoperative quality of life. A review of the international literature concerning coronary, aortic and mitral valve surgery in eighty-year-olds suggests that the postoperative complication rate, the duration of intensive care and the early postoperative mortality are significantly higher than in younger subjects. However, the long-term survival probability is similar to that of a population of eighty-year-olds without heart disease and significantly better than after non-surgical treatment. The quality of life is usually improved by the operation as suggested by a significant reduction of the symptoms and improvement of functional capacities. The favourable long-term results of cardiac surgery in eighty-year-olds therefore compensate for the higher intraoperative risk and argue in favour of earlier intervention. PMID- 12555359 TI - [Limits of coronary angiography]. AB - Coronary angiography is the "gold-standard" of the diagnostic approach in the patients with suspected coronary artery disease. Although it can be safely and easily performed in a modern cardiac catheterization laboratory, it is nevertheless responsible for a certain degree of morbidity and mortality related to the invasive nature of the procedure, and especially to the use of an iodinated contrast agent. Unlike certain functional tests, the results of coronary angiography are extremely "operator-dependent", which probably constitutes the most difficult limiting element to overcome in practice. Prognostic elements provided by coronary angiography (site and extent of lesions) must also be compared with the results of the reference functional test, Thallium 201 stress myocardial scintigraphy. PMID- 12555360 TI - [Diagnostic value of endocoronary echography]. AB - Intracoronary ultrasound allows in vivo examination of the various layers of the wall of arteries affected by atherosclerosis. Its use in the catheterisation laboratory has therefore allowed better understanding of the mechanism of the action of the various angioplasty procedures, allowing better guidance of the indications and better control of these techniques. It also helps to guide these procedures, thereby facilitating an optimal immediate result, which determines the long-term outcome. The characteristics of intracoronary ultrasound (high resolution, proximity of the arterial wall, echogenicity of constituents of the plaque) make it the instrument of choice to study the composition and structure of the plaque, which determine the clinical presentation and active potential, especially the risk of complications (rupture). The development of image and crude acoustic signal processing techniques have improved the performance of the analysis of these documents and raise hopes of one day being able to predict and anticipate plaque rupture. PMID- 12555361 TI - [Study of the efficacity and tolerance of diltiazem LP 300 mg in 2000 hypertensive patients (alone or combined with an angiotensin converting enzyme inhibitor]. PMID- 12555362 TI - [New perspectives in the treatment of hypertension]. AB - Very interesting results of several long-term studies concerning the treatment of hypertension were recently presented at the 18th "Journees de l'Hypertension" conference. Two of them are particularly interesting as, to our knowledge, they are the first studies designed to evaluate the use of fixed very low dose ACE inhibitor/diuretic bitherapy as first-line treatment for systolodiastolic hypertension and isolated systolic hypertension, over a long follow-up period. Both studies included hypertensive patients over the age of 65, and the only option left to the investigators was to adapt the dosage in the case of insufficient blood pressure control (suspine DBP > 90 mmHg). In particular, the results in the overall population show that 80% of patients "controlled" at 3 months, subsequently remained controlled throughout the study period (total of 15 months). The results obtained in isolated systolic hypertension were of the same order of magnitude. The results observed with this fixed very low dose ACE inhibitor/diuretic bitherapy appear very promising in terms of compliance with treatment and long-term blood pressure control, compared to those obtained with any other antihypertensive drugs indicated as first-line treatment of hypertension. PMID- 12555363 TI - [Medico-economic evaluation of abciximab]. AB - Abciximab (ReoPro) is a chimeric monoclonal antibody directed against platelet glycoprotein IIb-IIIa. It reduces the frequency of major cardiac ischaemic events after percutaneous transluminal coronary angioplasty (PTCA). This treatment is more expensive than the commonly used treatment (aspirin and heparin). However, a treatment can only be validly assessed in term of its cost-benefit ratio. The authors present the results of a medico-economic evaluation of abciximab in the French institutional context, based on the three-year clinical results of the EPIC study (Evaluation of abciximab for the Prevention of Ischemic Complications). The efficacy of treatment was expressed by means of two quantitative indicators: the number of patients with no major ischaemic event (MIE) and the number of years of life without myocardial infarction or revascularization. According to French good practice recommendations for economic evaluation of therapeutic strategies, major ischaemic events were assessed by using the complete cost of the corresponding homogeneous groups of diseases (GHM). Medico-economic evaluation of abciximab showed that the net excess cost per patient for national health insurance was 395 Francs, while the cost of the product is 5,300 Francs. The incremental cost per additional patient with no major ischaemic event is 6,585 Francs and the incremental cost per additional year of life without myocardial infarction or revascularization is 2,469 F. From a public health point of view, abciximab administered by bolus injection plus a 12-hour infusion, presents a good cost-effectiveness ratio in the prevention of cardiac ischaemic complications in high-risk patients after PTCA. PMID- 12555364 TI - [Double aortic arch]. PMID- 12555365 TI - [Mid-term prognostic value of plasma heavy-chain myosin in thrombolysed myocardial infarction]. AB - Plasma myosin heavy chain assay, which can be easily performed during the acute phase of myocardial infarction, is a recent method allowing quantitative assessment of the extent of infarction. However, to our knowledge, its prognostic value has not been studied in contrast with serum myosin light chain assay. We monitored the state of health of 40 patients (including 37 men with a mean age of 56 years) for two years after a first myocardial infarction, thrombolized during the acute phase. Their survival (mortality) and the development of "cardiac events" (MI, angina, sudden death, etc.) were evaluated at 2 years. The results observed at 2 years were correlated with the initial plasma myosin assay results and other direct and indirect methods of assessment of the extent of infarction, performed during the acute phase of myocardial infarction (cardiac enzymes, contrast angiography). The main result of this study is the demonstration that an unusual plasma myosin release kinetic (complex appearance) is predictive for the medium-term development of heart failure (p = 0.04) and/or destabilization of coronary insufficiency (p = 0.02). These results need to be emphasized, as with only 5 serum myosin assays performed over a 10-day period, it seems possible to identify a group of patients at high risk of medium-term complications, who possess a complex release kinetic during the acute phase of myocardial infarction and a value for area under the curve greater than 10.470 microliters U/L (cut-off value, p = 0.043). PMID- 12555366 TI - [Evaluation of arrhythmic risk in coronary insufficiency]. AB - Half of all deaths occurring in patients with heart failure are sudden deaths probably related to a malignant ventricular arrhythmia. The pathophysiological mechanisms of these arrhythmias are unclear, but left ventricular function, hypokalaemia accentuated by diuretics and treatments altering inotropism play a definite role. Because of the diversity of aetiologies generating heart failure, the multiplicity of fatal arrhythmias and the multifactorial origin of these arrhythmias, there is no formal marker for the risk of sudden death in patients with heart failure, at the present time. In addition to the NYHA classification and detection of episodes of syncope, assessment of these patients must be as complete as possible, at least including repeated evaluation of the ejection fraction, Holter ECG monitoring and detection of delayed ventricular potentials. PMID- 12555367 TI - [Value of arterial rigidity measurements in cardiovascular pathology]. PMID- 12555368 TI - [Indication for cardiac stimulation in minimally symptomatic bradycardias]. PMID- 12555369 TI - [Contrast echography. A new approach to myocardial perfusion]. PMID- 12555370 TI - [Evaluation of results and strategic choices in Doppler intracoronary angioplasty]. PMID- 12555371 TI - [Blood pressure parameters and cardiovascular risk in the elderly]. AB - The treatment decision must take into account the benefit and risks related to the intervention: the benefit demonstrated and quantified in many therapeutic trials in hypertension in the elderly, but also the patient's initial risk. It is now recognized that, in elderly hypertensive patients, systolic blood pressure is a better predictor of morbid and lethal events related to hypertension than diastolic blood pressure. Recent data in the medical literature attribute a predictive role to pulse pressure which is even greater than that of systolic blood pressure. From a pathophysiological point of view, the level of pulse pressure reflects the degree of rigidity of large arterial trunks. The arterial rigidity parameter could integrate the harmful effect of "cardiovascular risk factors" (hypertension, but also atherogenic dyslipidaemia, diabetes, smoking, homocysteine, genetic factors, etc.) on the years or decades of exposure, and pulsed pressure would therefore appear to be a better marker of cardiovascular risk than other blood pressure parameters. Pulse pressure should therefore be integrated into the benefit/risk ratio of antihypertensive treatment in the elderly. PMID- 12555372 TI - [Blood pressure variability in the elderly]. AB - In the elderly, the difference in blood pressure (BP) between the day and the night is less marked than in young subjects, but it is also more unstable and more sensitive to changes in position, meals, and physical activity. A deficient cardiac baroreflex appears to be involved in this phenomenon. Mayer's oscillation, usually considered to reflect sympathetic vascular activation, is not amplified by standing, but generates slower oscillations, at lower frequencies. The sensitivity of the cardiac baroreflex is inversely correlated with age, but, after the age of 60 years, the correlation with the level of systolic blood pressure (SBP) is greater than with age and accentuation of SBP is the main determinant in problems of postural hypotension. Hypertension and alterations in the afferent pathway of the baroreflex, adrenergic receptors, structure of the arterial wall, and intravascular volume appear to participate in the changes of blood pressure variability observed in the elderly. PMID- 12555373 TI - [Left ventricular consequences of hypertension in the elderly]. AB - Modifications of left ventricular morphology observed during ageing are very similar to those encountered during hypertension, as if hypertensive patients present premature ageing of their cardiovascular system. Hypertension in the elderly amplifies the concentric remodelling and alteration of diastolic function observed in normotensive subjects during ageing. The main determinant of these modifications of morphology and cardiac function is the increased rigidity of large arterial trunks with age. Treatment of hypertension therefore represents one of the essential ways to control this pathological ageing. PMID- 12555374 TI - [How to diagnose hypertension in the elderly?]. AB - The management of hypertension in the elderly requires careful diagnosis, which is not always easy in this population because of the increased variability of blood pressure. The practitioner is faced with three questions in this diagnostic approach: how to evaluate the patient's real blood pressure level, how to assess the degree of cardiovascular impairment and concomitant disease and finally how to evaluate the subject's global cardiovascular risk. Self-monitoring of blood pressure, which allows measurement of blood pressure by the patient himself in his usual environment, provides a better assessment of the elderly patient's real blood pressure level. PMID- 12555375 TI - [Antihypertensive treatment and prevention of dementia]. AB - Hypertension is a risk factor for dementia, particularly vascular dementia. The objective of the "Vascular Dementia" project of the European multicentre Syst-Eur trial was to study the influence of antihypertensive therapy on the incidence of dementia in elderly patients with isolated systolic hypertension included in this trial. Patients were over the age of 60, with systolic blood pressure between 160 and 219 mmHg and diastolic blood pressure less than 95 mmHg in the sitting position and were not demented. After randomization, they were treated under double-blind conditions with nitrendipine associated with or replaced by enalapril or hydrochlorothiazide or an equivalent placebo. Cognitive functions were evaluated by the Mini-Mental State test (MMS) on inclusion then annually. When the MMS score was < or = 23, a complementary assessment was performed to establish the diagnosis of dementia according to DSM III-R criteria. The Modified Ischaemia score specified the aetiology of dementia. A total of 1180 and 1238 subjects were randomized to the placebo and active treatment groups, respectively. They presented comparable characteristics on inclusion. The median intent to treat duration of follow-up was 2.0 years. The incidence of dementia was decreased by 50%, from 7.7 per thousand patient-years in the placebo group to 3.8 per thousand patient-years in the active group (p = 0.05). There were 15 and 8 cases of dementia of the Alzheimer type, 4 and 3 cases of mixed dementia, 2 and 0 cases of vascular dementia in the placebo and active treatment groups, respectively. In conclusion, in subjects over the age of 60 years with isolated systolic HT, antihypertensive treatment initiated with nitrendipine can reduce the incidence of dementia. PMID- 12555376 TI - [Benefits of hypertension treatment in the elderly]. AB - The benefits of treatment of hypertension in the elderly have been demonstrated by the results of therapeutic trials conducted in subjects over the age of 60. In 1985, the EWPHE trial, which demonstrated that thiazide diuretics decreased the main cardiovascular complications observed in elderly hypertensives, opened the way to the demonstration of the preventive efficacy of other classes of antihypertensives. The high level of cardiovascular risk observed in the elderly explains why prescription of antihypertensives provides a greater benefit in elderly hypertensives than in younger hypertensives. In hypertensive patients over the age of 80, the benefits of drug-induced blood pressure reduction have been demonstrated in terms of prevention of nonfatal complications. PMID- 12555378 TI - [Evolution of health care in France: comparison elements with foreign countries]. AB - This article reviews the financial growth reports of five health systems: France, Germany, United Kingdom, United States and Canada. It is designed to provide descriptive elements related to expenditure: medical demographics and health care professionals in general, hospital densities, availability of sophisticated medical equipment, medical approach to evaluate the needs for specialists, responsible for ordering expensive technologies (especially cardiologists). The results are presented in summary table form and show that debates on health system reforms too often forget these basic descriptive elements able to diagnose a crisis situation in the system, whether it is financial and/or social and consequently the possible remedies than can be applied. Finally, these discussions also often forget the significantly different epidemiological situations observed between rich countries, as illustrated by the situation of cardiovascular diseases in the countries used as examples. In conclusion, the data presented provide a basis for reforms of health systems, which have been converging since the beginning of the 1990s: closure of short-stay hospital beds, control of medical demographics, specialties and technical procedures, attempts to take the morbidity of the population into account. PMID- 12555377 TI - [Brain natriuretic peptide (BNP) in coronary insufficiency: relationship with left ventricular filling and exercise tolerance]. AB - Brain natriuretic peptide (BNP) is a recently discovered peptide, secreted by the atria and ventricles in response to parietal distension. It was recently proposed as a screening test for left ventricular failure. The authors assayed this peptide at rest in 37 patients with chronic heart failure due to left ventricular systolic dysfunction and another 20 patients with various diseases (respiratory failure, cirrhosis, heart transplantation, "diastolic" heart failure) but normal left ventricular systolic function. A significant increase compared to normal values was observed not only in the group of heart failure patients, but also in patients with all other diseases. BNP was significantly higher in NYHA class IV patients. The relationship between plasma BNP levels and ejection fraction was not significant. On the other hand, a good correlation was observed between BNP and left ventricular filling parameters evaluated by cardiac Doppler: E wave deceleration time (r = -0.53, p = 0.001), E/A ratio: r = 0.57 p = 0.005) or VO2 max (r = -0.55, p < 0.005). PMID- 12555379 TI - [Homocysteine and coronary events in coronary disease patients]. AB - The objective of this study was to determine the prognostic value of serum homocysteine levels in patients with coronary heart disease. Homocysteine was assayed in 76 coronary patients with a mean age of 59.2 years hospitalized for myocardial ischaemia or myocardial infarction. Percutaneous transluminal angioplasty was performed in 47 (70%) of these patients during this hospitalization. The mean follow-up for these patients was 22 months (range: 11 to 67 months). In these patients, serum homocysteine levels were not correlated with the usual risk factors of coronary heart disease (age, sex, treated hypercholesterolaemia, smoking, diabetes) except for hypertension. It was strongly correlated with serum creatinine (R = 0.61; p = 0.0001). Eleven patients presented a major event during follow-up (8 deaths, 1 nonfatal myocardial infarction, 1 cardiac transplantation) and 16 underwent a revascularization procedure. The blood homocysteine level does not have any prognostic value for any coronary events. However, it is higher in patients who develop a major event than in those which do not (15.8 +/- 4 mumol/l versus 11.5 +/- 6.6 mumol/l, p = 0.05). Using multivariate analysis, taking into account age, serum creatinine and serum homocysteine, only serum homocysteine was predictive of major event-free survival (p = 0.02). PMID- 12555380 TI - [Role of the Na+/H+ pump in heart ischemia-reperfusion lesions in the neonatal period]. AB - OBJECTIVE: Amiloride 5-(N,N-dimethyl)(AMD), an Na+/H+ pump inhibitor, was used to evaluate the maturity and sensitivity of the Na+/H+ pump in neonatal myocardial cells. METHOD: Using the isolated contraction isometric heart model, the hearts of neonatal rabbits (10-12 days) were stopped by cardioplegia with and without AMD (20 mumoles/l), then submitted to 40 minutes of ischaemia at 37 degrees C or 120 minutes of ischaemia at 20 degrees C before being reperfused for 30 minutes at 37 degrees C. When AMD was added to the cardioplegic solution, it was also added (20 mumoles/l) to the Krebs solution for the first five minutes of reperfusion. Functional assessment of myocardial contractility and relaxation was performed before aortic clamping and after 30 minutes of reperfusion. Coronary venous drainage during reperfusion was collected to evaluate CPK levels. RESULTS: To 37 degrees C or 20 degrees C, Na+/H+ pump inhibition was marked by improvement of all myocardial contractility and relaxation parameters at the end of the reperfusion period. A significantly reduced CPK release wase also observed during reperfusion associated with Na+/H+ pump inhibition. CONCLUSION: This study demonstrates maturation of the Na+/H+ pump in neonatal myocardial cells and confirms the role of this ion exchange pump in the pathogenesis of ischaemia reperfusion lesions. The study at 20 degrees C demonstrated the sensitivity of the Na+/H+ pump to a low transmembrane proton gradient. PMID- 12555381 TI - [Aorto-pulmonary collaterals in an infant with 22q11 monosomy]. AB - The authors present the case of a young girl in whom the diagnosis of pulmonary atresia with ventricular septal defect and multiple aorto-pulmonary collateral vessels was established in the context of microdeletion of chromosome 22. Surgical treatment was excluded following the discovery by angiography of the absence of central pulmonary arteries, a multifocal pulmonary collateral circulation and the appearance of multiple peripheral pulmonary artery stenoses on various aorto-pulmonary collateral vessels during the first months of life. This case illustrates the hypothesis recently developed in the literature according to which the chromosomal abnormality is responsible not only for an anomaly of conotrunkal septation, but also early disorders of branching of the pulmonary vascular bed and disturbances of the connection of peripheral pulmonary segments to central arteries, which are responsible for severe pulmonary vascular abnormalities making biventricular repair of this heart disease in children with 22q11 microdeletion almost always impossible. PMID- 12555382 TI - [Werner's disease presenting as ischemic mitral incompetence]. AB - Werner's syndrome or progeria, described for the first time in 1886, is a rare disease with autosomal recessive transmission, characterized by premature ageing of connective tissues. About 200 cases have since been reported in the literature. Most patients die young, generally from heart failure due to early coronary atherosclerosis [1]. The authors report the case of a 46-year-old woman presenting with cardiovascular abnormalities, unusual for her age, associated with a particular morphotype belonging to Werner's syndrome. PMID- 12555383 TI - [Treatment of coronary insufficiency in diabetics. Part 3: chronic coronary insufficiency]. AB - The drug treatment of chronic coronary insufficiency in diabetic patients is now well defined. Platelet antiaggregants, especially aspirin, must be prescribed in the long-term or even indefinitely. Other drugs (beta-blockers, calcium channel blockers, nitrates, etc.) can be used in the same way as in the absence of diabetes. Angioplasty gives immediate favourable results in diabetics, very similar to those obtained in the absence of diabetes. In contrast, the longer term prognosis is less favourable, as the mortability, myocardial infarction, restenosis and bypass graft rates are significantly higher. First-line stenting lowers the restenosis rate to a level comparable to that observed in non diabetics. However, instrumental revascularization is less complete than surgical revascularization and the number of redilatations and/or secondary bypass grafts remains high. The indications, mortality and early complications of coronary surgery are now identical to those observed in the absence of diabetes. Its long term results are significantly more favourable than those of medical treatment or even angioplasty, although this issue is still controversial. The improved prognosis observed in operated diabetic coronary patients is due to the more frequent use of arterial bypass grafts. The maintenance of blood glucose control and correction of the frequently associated cardiovascular risk factors (obesity, sedentary lifestyle, smoking, HT, dyslipidaemia) increase the efficacy of treatment of coronary insufficiency in diabetic patients. This goal can only be achieved by permanent, unfailing collaboration between cardiologists and diabetologists. PMID- 12555384 TI - [Effect of diltiazem on lipid profiles and heart rate in hypertensive patients]. AB - Diltiazem is an antihypertensive belonging to the class of heart rate-slowing calcium channel blockers; its antihypertensive efficacy has been demonstrated for a long time. Controlled trials have also demonstrated that it does not interfere with lipid parameters. This open multicentre trial was designed to verify the absence of any action of diltiazem on lipid parameters as well as its effect on the heart rate of hypertensive patients. The patients included had to take one capsule of diltiazem 300 mg sustained release formulation, developed by Laboratoires Synthelabo, in the morning for 6 months. A lipid assessment and an electrocardiogram were performed before and at the end of the trial. One hundred and forty-six patients were included: 81 underwent a complete lipid assessment (7 parameters) before and after treatment and 69 underwent an electrocardiogram at rest to the same study times. Total cholesterol (p = 0.0007), LDL cholesterol (p = 0.005) and apoprotein A1 (p = 0.0002) were significantly decreased. Blood pressure decreased progressively throughout the study and was "controlled" (supine diastolic blood pressure < or = 90 mmHg) after six months of treatment in 77% of patients. Heart rate decreased significantly at the end of the study; no pathological variation of PR, QRS and Qt intervals was observed. The effect on heart rate, also studied in patient subgroups defined by their baseline values, was more marked when the baseline heart rate was increased. The investigators considered the safety to be good at each visit, in 80% of cases. Sixteen patients dropped out of the trial because of adverse effects or intercurrent disease. The adverse effects most frequently reported were oedema of the lower limbs (7.5%), headache (4.8%) and hot flushes (2.7%). No harmful effect of diltiazem on lipid parameters was observed during this open trial; some lipid parameters even decreased at the end of treatment. Clinically, diltiazem lowered blood pressure and decreased heart rate. The reduction of heart rate was more marked in the case of high baseline heart rate. Safety was satisfactory. PMID- 12555385 TI - [Heart insufficiency]. PMID- 12555386 TI - [Epidemiology of heart insufficiency]. AB - The epidemiology of congestive heart failure is not well known in France, despite its dramatic economic impact on the health care system. The number of patients with congestive heart failure is about 500,000 with 120,000 new cases per year. The incidence ranges from 4@1000 in men and 3@1000 in women 55-64 years old to 50@1000 in men and 85@1000 in women 85-94 years old. Mean age at onset is 73.5 years, and two thirds of the patients are older than 70 years of age. There are about 3,500,000 outpatient visits and 150,000 hospitalizations for congestive heart failure every year. It is responsible for more than 32,000 deaths every year. Costs of congestive heart failure represent more than 1% of the total medical costs. It represents a major and growing public health burden. This should encourage us to optimize its medical treatment and prevention. PMID- 12555387 TI - [Genetic myocardiopathies: present and future]. AB - Heart failure is a major health problem and is associated with a high mortality and morbidity. Recently, the role of the genetic background in the onset and the development of the disease has been evidenced in both heart failure with and without systolic dysfunction, and in familial and non familial forms of this condition. Several genes and loci are know identified as responsible for dilated cardiomyopathies and for hypertrophic cardiomyopathies in familial and monogenic forms. Susceptibility genes and modifier genes are also studied in nonfamilial forms of dilated cardiomyopathies. The analysis of genetic factors that predispose to heart failure looks promising. It should allow to better understand the underlying mechanisms that promote the development and the progression of the disease, to identify subjects at risk for the disease who would benefit of an early medical management and promote the development of pharmacogenetics. PMID- 12555388 TI - [Prognostic factors of chronic heart insufficiency]. PMID- 12555389 TI - [Medical treatment of chronic heart insufficiency]. AB - The treatment of chronic heart failure by alteration of systolic function has made remarkable progress over the past ten years; this progress has been linked to the results of large-scale clinical trials involving tens of thousands of patients. The medical approach to heart failure is therefore based on established clinical data. The therapeutic aims have been extended: not only should treatment relieve the symptoms of the disease, but also reduce the frequency of relapse and hospitalization, and prolong survival duration. At the onset of the third millennium, three therapeutic classes have emerged: conversion enzyme inhibitors, spironolactone, and beta-blockers, which although counter-indicated only a short while ago, now constitute the revolutionary aspect of the approach to treatment of chronic heart failure. In addition to drug therapy and as in the case of other chronic diseases, the significant role of patient awareness, i.e., the information and education that is made available to the individual regarding his or her illness, should also be taken into consideration. Regarding the latter aspect, the experience gained in this field by the care systems can be utilized. However, recent epidemiological data have shown that the advances made in the treatment of heart failure only concern a minority of patients; and that elderly subjects, women, and patients suffering from heart failure in spite of normal systolic function, which constitute the majority of cases, have not been considered in these trials. PMID- 12555390 TI - [Drug prospects in the treatment of heart insufficiency]. AB - Pharmacological and validated treatment of chronic heart failure (HF) includes successively angiotensin converting enzyme inhibitors (ACEi), beta-blockers and antialdosterone, which is associated with diuretics. The effectiveness of this manner in which to block more and more hormonal systems demonstrate the validity of the "hormonal" paradigm to explain heart failure. Therefore broader educational means are required to increase the prescription of these drugs for HF. Several questions about these drugs remain unresolved: HF with preserved systolic function and elderly patients, class effect, and the role of antagonists of angiotensin II receptors (as an alternative or associated with ACEi). Other short- and mid-term pharmacological perspectives target target hormonal systems and cytokines: endothelin-receptor antagonists, inhibition of natriuretic peptide degradation (via neutral endopeptidase), and newer drugs acting against TNF such as etanercept. Moreover, recent knowledge about molecular mechanisms of myocardium remodeling allows new drug strategies with target more specifically remodeling such as metalloproteinases. Finally, these perspectives should be largely modified by on-going research in the field of genomics. PMID- 12555391 TI - [Cardiac stimulation and heart insufficiency]. AB - Dilated cardiomyopathies are frequently associated with atrioventricular and intra ventricular conduction disorders. Such conduction disorders modify the cardiac activation sequence and impair left ventricular systolic and diastolic function. The aim of pacing in dilated cardiomyopathy is to restore a cardiac activation sequence as normal as possible. Conventional dual-chamber pacing was first proposed to treat drug-refractory heart failure but was soon replaced by multisite biventricular pacing. The aim of biventricular pacing is to correct intraventricular asynchrony by pacing the two ventricles simultaneously. Acute studies have shown that biventricular pacing could improve hemodynamic parameters in patients with dilated cardiomyopathy and severe intraventricular conduction disorders. More recently, chronic studies demonstrated a significant improvement in exercise tolerance and quality of life with biventricular pacing in patients with drug-refractory heart failure and with intraventricular conduction disorders. To safely pace the left ventricle specific materials and tools had to be developed. With those developments, biventricular pacing is now a feasible and safe technique. Regarding the results of several clinical studies, biventricular pacing could now be proposed to treat patients with drug-refractory heart failure in the setting of chronic left ventricular systolic dysfunction and with major intraventricular conduction disorders. PMID- 12555392 TI - [Surgery of heart insufficiency]. AB - Apart from heart transplantation for heart failure, the problem arises of which surgical approach should be adopted to treat this disorder. Ischemic heart disease can be surgically managed in three ways: via bypass, the Dor procedure, or by isolated or associated mitral plasty; however, cases of enlarged heart disease can only be surgically treated in two ways: i.e., by mitral plasty, or by the Batista procedure. In cases of ischemic heart disease, the following conditions must be present for coronary bypass: the patients should have an adequate contractile myocardial reserve, that is to say the left ventricle should not be greatly enlarged (< 80 mm in telediastole) or a cardiac output reserve, and there should not be any sign of over-high pulmonary hypertension (an index of > 1.6 or a pulmonary pressure of < 45); an assessment of myocardial viability should then be carried out, mainly based on a thallium fixation at rest and on echographically determined doubtamine-associated stress. In the present study, the mortality rate in a series of 260 patients was 6.3% for subjects aged under 70 years old, with an actuarial survival rate of 82% at one year post-surgery, and of 70% at five years. The Dor procedure can be used in the treatment of dyskinesia, which is now practically non-existent, but also in cases of acute akinesia with resulting left ventricular dysfunction. The aim of this technique is to alter the form of a cavity that has become ovoid to an elliptical form via the insertion of a circular endoventricular patch. The results reported for this technique show an improvement in functional class and ejection fraction. Finally, the technique for repairing mitral failure is more complicated than the two previous methods, as it requires a dynamic assessment of mitral failure, which is best carried out by an evaluation of echographically determined stress. Any mitral failure of ischemic origin of > grade 2 can be corrected during bypass surgery by ring insertion, thereby effecting a simple annuloplasty. On the other hand, the assessment of cases of enlarged heart disease is more complicated, and it is more difficult to carry out palliative surgery. The mitral plasty procedure proposed by Bolling is the technique of choice for patients with severe mitral failure, in general when the ventricle is not too enlarged. However, surgery involving the reduction in size of the left ventricle (the Batista procedure) always includes mitral plasty, and may be performed in patients with a very enlarged ventricle (> 70 mm), in general with moderate mitral failure. These two techniques have been critically assessed both as regards results and when they should be adopted, and their limitations have also been discussed. In conclusion, there are valid surgical alternatives to heart transplantation in cases of heart failure that does not respond to medical treatment, and they should probably be seriously considered before any decision is made to perform heart transplantation. These results appear encouraging, particularly in terms of functional class and left ventricular function, but there are conflicting results for hemodynamic improvement. As regards survival, it is not yet possible to propose prospective randomized trials to compare medical treatment with these surgical techniques. However, further development of these techniques is bound to occur, and an ever-widening gap will exist between the limited number of cases requiring transplantation and the more complex surgical approaches adopted in future, such as permanent circulatory backup or xenografts. PMID- 12555393 TI - [Role of rehabilitation in the treatment of chronic heart insufficiency]. AB - Rehabilitation is an important component of the modern comprehensive care plan for patients with chronic heart failure. Cardiac rehabilitation combines exercise training with therapeutical adaptations, behavioral modifications and psychosocial interventions. Based on these data, patients with controlled heart failure should be involved in cardiac rehabilitation programs. Training prescription needs a strict previous cardiac evaluation. Exercise training monitoring must be adjusted to the physical tolerance of each patient. Cardiac rehabilitation has been found to improve functional capacity, reduce symptoms, and finally reduce cardiac morbidity and mortality. These beneficial effects were associated with muscular, endothelial and ventilatory improvements. Reduced sympathetic tone may decrease arrhythmias and may limit the progression of left ventricular dysfunction. PMID- 12555394 TI - [Taxonomic study of four isolates of Saccharomonospora]. AB - Four strains, 19.1, 22.1, 202, and 212, were isolated from the soil samples of Guangxi province in China. Basing on their morphological and cell chemical characteristics(cell wall type IV, sugar type A, without mycolic acids), they were circled into the genus of Saccharomonospora. They were studied with 7 type strains of the 5 currently recognized species in the genus. The results of Ribotyping suggest that strain 19.1 blongs to the species of S. viridis, strain 202 is a member of S. caesia, strain 22.1 and strain 212 are the same species, which is different to the 5 currently known species. It is closely related to S. viridis. However, the further classification of these two strains at species level needs 16S rRNA sequence date. PMID- 12555395 TI - [Construction of full-length cDNA clone and complete nucleotide sequence of RNA2 of cucumber mosaic virus SD strain]. AB - The full-length cDNA clone of CMV-SD RNA2 was constructed from two cDNA clones, which were obtained by RT-PCR with primers synthesized based on terminal sequences determined using 5' RACE and 3' RACE, respectively; and the complete sequence was determined. It consists of 3048 nt and there are two partial overlapped ORFs. ORF1, which is located in the 5' portion (79-2652 nt), codes for 2a protein of 858aa, and GDD consensus sequence reported to be "polymerase site" was found in deduced 2a protein sequence. ORF2, which is located in the 3' portion (2414-2746 nt), codes for 2b protein of 111aa. The overall nucleotide sequence homologies with RNA2 of CMV Fny and Q strain, which are representatives of subgroups I and II, respectively, are 91.7% and 75.6%; the amino acid sequence homologies are 93.8% and 67.7% for 2a protein, and 83.0% and 51.3% for 2b protein, respectively. These results demonstrated that CMV-SD belongs to subgroup I. PMID- 12555396 TI - [Deletion analysis and functional studies of the promoter from commelina yellow mottle virus]. AB - Commelina Yellow Mottle Virus(CoYMV) is a double-stranded, circular DNA virus and its promoter could direct GUS gene specifically expressing in phloem tissue of transgenic tobacco plants. To determine the optimal promoter sequence for pholem specific gene expression, CoYMV promoter was deleted from its 5 end to form promoter fragments with 5 different lengths. Chimeric GUS genes were constructed using the promoter deletion based on the binary vector pBI121. Transgenic tobacco plants evidenced by PCR analysis were obtained with each kind of chimeric GUS gene structure by Agrobacterium mediated transformation. The results of GUS activity assay and histo-chemical staining showed that most of the chimeric GUS genes were expressed in transgenic plants. The GUS activity with the promoter deleted to -870 bp was about 78% higher than that of the full length promoter(1040 bp) and was a little higher than that of the promoter deleted to 585 bp, but the difference is not significant. The GUS activity reduced significantly when the promoter was deleted to -447 bp or -232 bp, whereas the property of phloem-specific expression pattern was still retained. When the promoter was deleted to -44 bp, just upstream adjacent to the TATA box, its tissue-specificity was lost and the activity was reduced to undetectable level. These results suggest that the region between -870 bp-232 bp and downstream of 232 bp of CoYMV promoter could be responsible for promoter activity and tissue specific expression, respectively. A negative regulation sequence might exist upstream of -870 bp of the CoYMV promoter. Therefore, we recommend that the optional CoYMV promoter sequence for phloem specific expression could be downstream from -870 bp or -585 bp. In comparison with CaMV 35S promoter, the GUS activity when driven by -870 bp CoYMV promoter was about 70% of that when driven by the 35S promoter. Considering the fact that 35S promoter-GUS gene is constitutively expressed, while the CoYMV promoter-GUS gene is expressed only in phloem tissues, the activity of the latter in phloem may be the same with or even higher than that of the 35S promoter. PMID- 12555397 TI - [Cloning and sequencing of cDNA from G protein gene of subgroup B respiratory syncytial virus strain isolated in China]. AB - The nucleotide sequence of the G protein gene of respiratory syncytial virus (RSV) CC169 strain isolated from China that has been identified as subgroup B with monoclonal antibodies, was determined from cDNA that had been amplified by RT-PCR and cloned into pTZ18R plasmid vector. The homology of nucleotide was 94% as compared with G protein cDNA of a RSV prototype strain (CH18537). Deduced amino acid identity of G protein was 89.4%. The amino acid changes were only in the extracellular part of the protein where there were two extensive divergent domains with a highly conserved region in between; whereas the cytoplasmic and transmembrane domains were conserved. This study demonstrates the sequence diversity of the G protein of subgroup B RSV between a Chinese isolate and the prototype strain CH18537. PMID- 12555398 TI - [Cloning and expression of the binary toxin genes of Bacillus sphaericus C3-41 in a crystal minus B. thuringiensis subsp. israelensis]. AB - Bacillus sphearicus C3-41, belonging to serotype H5a5b, is ahighly toxic strain isolated from a mosquito-breeding site in China. It has been shown that it had a higher toxicity against Culex spp. than the reference strain 2362 at the laboratory and field conditions. Using synthetic oligonucleotides designed on the basis of the binary toxin gene sequence of 2362, a 1.1 kb DNA fragment was produced and the genetic library was prepared from a HindIII digest of total DNA from C3-41. One colony containing the 3.5 kb HindIII fragment was selected for further studies. Sequence analysis revealed that this 3.5 kb Hind DNA fragment was composed of 3479 nucleic acids and the sequence of the binary toxin gene of C3-41 is completely identical to that of strain 2362. The toxin genes have been transferred into a nontoxic crystal-minus strain of Bacillus thuringiensis subsp. israelensis. The recombinant strains could express the binary toxin of B. sphaericus as crystaline structures during their sporulation. The expression products of the recombinants have a highly toxicity to susceptible Culex pipiens subsp. quinquefasciatus and no toxicity to resistant larvae(Culex pipiens subsp. pipiens) from France. PMID- 12555399 TI - [Cloning and sequencing analysis of rice stripe virus genome segment 4 of Chinese isolate Y]. AB - The cDNA fragment covering full-length sequence of RStV RNA4 of Yunnan isolate in China was obtained by RT-PCR. The PCR-derived fragment was then cloned into vector pCRII. The cloned cDNA was sequenced. Comparison of the nucleotide and deduced amino acid sequences with those of the Japanese isolate T was made. The results showed that at the nucleotides level, vORF, vcORF and the intergenic region had 94.9%, 94.1% and 86.1% identity respectively, the 5'-untranslational region was exactly the same as that of Japanese isolate T, while the 3'-terminal sequence had 96.1% identity, differing by two nucleotides; at the amino acid level, vORF and vcORF had 99.4% and 98.3% identity respectively. Therefore, as well as being exactly the same size for the two isolates, the amino acid sequences of the coding regions and the 5'- and 3'-terminal sequences were well conserved. Our results indicated that the Chinese isolate is closely related to the Japanese isolate T. PMID- 12555400 TI - [Using lux genes marker technique to track Pseudomonas chlororaphis PL9L in cotton rhizosphere]. AB - Tn7-luxCDABE marker system was successfully transferred into Pseudomonas chlororaphis (strain PL9) by means of transformation and conjugation and a stable lux-marked strain PL9L was obtained. The colonizing dynamics and distribution of the luminescent bacteria PL9L in the rhizosphere of cotton planted in pots and rhizoboxes were studied by the methods of X-ray film imaging and enumeration of luminescent colonies on agar media, The results of pot culture experiment showed that PL9L successfully colonized in the rhizosphere of cotton. In pot cultures of sterile soil the highest colonizing level(3.1 x 10(2) cfu/g root soil) was reached on 6th day after seeds sown; On 56th day, the population of PL9L tended to stable and decreased to 1.7 x 10(9) cfu/g root soil) but in pot cultures of unsterile soil, the highest colonizing level(1.1 x 10(9) cfu/g root soil) was reached on 8th day. On 46th day, the population of PL9L tended to a stationary state, the numbers of them were 1.4 x 10(2) cfu/g root soil. The results of rhizobox culture experiment showed that PL9L spread from seeds toward the direction of root tip, but not synchronized with the stretch of roots. 6 days after seeds sown, in rhizobox culture of sterile soil, PL9L spread 12.0 cm below seeds, but in non-sterile soil was 11.0 cm. In the region of cotton root tip, PL9L were not detected. PMID- 12555401 TI - [Expression of xylose isomerase gene(xylA) in Saccharomyces cerevisiae from Clostridium thermohydrosulfuricum]. AB - The Clostridium thermohydrosulfuricum xylA gene encoding xylose(glucose) isomerase was cloned in the yeast expression vector pMA91 under the control of the PGK promoter, resulting in pBX-1, and transformed into Saccharomyces cerevisiae. Production of recombinant xylose isomerase was seen in the a Coomassie stained SDS-PAGE gel and the molecular mass was estimated to be 43 kD. The recombinant xylose isomerase showed the highest activity at 85 degrees C and pH7. The specific activity under these condition was 1.0 U/mg protein. At 30 degrees C and 40 degrees C, the relative activity was reduced to 3.7% and 11%, respectively, of the maximum. PMID- 12555402 TI - [Purification and properties of human lysozyme in engineered bacterium E. coli]. AB - To obtain the SDS-PAGE-pure human lysozyme, the crude enzyme of engineered bacterium E. coli was purified by chromatography on cation ion exchange of Express-Ion S. The optimum reaction temperature and pH of this lysozyme were 45 degrees C and 6.5, respectively. The isoelectric point is pH 8.91, and Km of the enzyme for Micrococcus lysodeikticus is 0.0311 mg/mL. The thermal stability of the engineered enzyme is more sensible than hen egg white lysozyme and human milk lysozyme. The sequence of 5 amino acids in N-end is same as designed, except an Met at the first. The affects of some metal ion on this enzyme were shown. Cu2+ of 0.01 mmol/L could completely inactivate the enzyme. PMID- 12555403 TI - [Production of neutral beta-mannanase by Bacillus subtilis and its properties]. AB - Neutral beta-mannanase was produced by Bacillus subtilis BM9602 isolated from a soil. For this strain the neutral beta-mannanase was produced only when polysaccharides were used as a carbon source. Organic nitrogen source was superior to inorganic nitrogen source on the enzyme production. The optimum liquid medium consisted of 4% konjak powder, 1% each of beef peptone and yeast extract. The optimum culture conditions were initial pH 8.5, temperature 35 degrees C and cultivation time 36 h. Enzyme activity of culture filtrate to 0.5% galactomannan polysaccharide was 96 IU/mL per minute at pH 6.0 and 50 degrees C for 10 min. The optimum pH and temperature for enzyme action were 6.0 and 50 degrees C respectively. The enzyme was stable below 50 degrees C and pH 5.0-10.0. The enzyme hydrolyzed konjak powder and locust bean gum to form oligosaccharides. PMID- 12555404 TI - [Strain screening and fermentation conditions of a novel heparinase-producing strain]. AB - The novel heparinase-producing bacterial strain Corynebacterium sp. was screened and isolated from soil. The optimum medium composition is (g/L): Trypticase 20, NaCl 1, K2HPO4 2.5, MgSO4 0.5, Heparin 2, maltose 20, pH 6.5. The optimum growth temperature was 27 degrees C while, maximum enzyme production was achieved at temperature 31 degrees C. When cultured at a rotating shaker at 30 degrees C for 24 hours, 200 r/min, 40 mL medium in 500 mL flask, the Production of heparinase reached 1700 u/L. PMID- 12555405 TI - [A new osmotolerant and glycerol-highly-producing species--Candida glycerolgenesis Zhuge sp. nov]. AB - The strain WL2002-5 isolated from natural samples and overproduced glycerol from glucose was identified systematically. The WL2002-5 fermented glucose and weakly fermented sucrose; assimilated glucose, sucrose, ethanol, weakly assimilated glycerol and citrate, did not assimilated nitrate, erythritol, arabinitol, mannitol. It grew well on the slopes containing 500 g/L of glucose or 10 mL/L acetate or at the temperature up to 40 degrees C. The minimum water activity for its growth was 0.890. It had a negative reaction with DBB and did vegetative reproduction by budding and easily formed Candida-type pseudohyphae but had no sexual reproduction. The molecular weight of its mitochondrial DNA was 20 kb. We conclude that WL2002-5 is a new species of Candida and nominated it Candida glycerolgenesis Zhuge sp. nov.. PMID- 12555406 TI - [Phase variation in Salmonella typhimurium phagetype]. AB - The phase variation of phagetype was observed in cultures of Salmonella typhimurium collected from Xinjiang Uygur Autonomous Region during 1980-1993. A few cultures of phagetype 4774 or 4776(full type) may change into another phagetype, i.e. 4000(phase 1) or 0774 and 0776 (phase 2). In reverse, more strains of these phagetype 4000 and 0774(0776) easily back into their original phagetype 4774 or 4776. But some 4000(phase 1) can change to 0776(phase 2), and some 0774(phase 2) can also change to 4000(phase 1). Similarly, a few cultures of phagetype 7776(full type) can change to 7000(phase 1) or 0776(phase 2). More strains of these phagetype 7000(or 7002) and 0776 reverse easily to their original phagetype 7776 or 7774. The variability is 15.6% from full type to phase 1 or phase 2, and is 53.2% from phase 1 or phase 2 to full type. Understanding of phase variation of Salmonella typhimurium phagetype will be helpful to Salmonella phagetyping, and will be significance on epidemiological analysis of Salmonella infection. PMID- 12555407 TI - [Establishment of transformation system of Streptomyces lincolnensis]. AB - Transformation system of S. lincolnensis was established to improve lincomycin- producing strains by Gene retransformation Technology. Under optimal conditions, 10(11) protoplasts were prepared from 25 ml SM culture medium, and the regeneration frequency of protoplasts was 16%. The transformation frequency of pIJ702 was up to 5.5 x 10(5)/microgram DNA. The results revealed that gene manipulation in S. lincolnensis was facile as that in S. lividans. PMID- 12555408 TI - [Taxonomy of the producing strain of agricultural antibiotic 2-16]. AB - In screening process of new agricultural, a streptomycetes, 2-16, was isolated from the soil sample collected in Huangsan. The secondary metabolites produced by the strain had strong inhibition to Sclerotinia sclerotiorum. By identification, this strain was regarded as new variety of Streptomyces ahygroscopicus and was called Streptomyces ahygroscopicus var. huangsanensis. PMID- 12555409 TI - [Studies on biosorption of Pb2+ by Phanerochaete chrysosporium]. AB - The biosorption of Pb2+ from solution by Phanerochaete chrysosporium was studied. The effects on biosorption capacity of pH, Pb2+ concentration in solution, co-ion and temperature, time have been examined. The adsorption capacity was compared among the biomass harvested at various growth stages. Pretreatment of biomass by NaOH solution improved its adsorption capacity. The uptake could reach 108.4 mg/g at optimal conditions of biosorption using pretreated biomass. PMID- 12555410 TI - [The HOG pathway and glycerol synthesis in yeast]. PMID- 12555411 TI - [Studies on DNA-DNA hybridization and 16S rDNA sequence of rhizobia isolated from Shapotou desert soil in Ningxia autonomous region of China]. AB - Based on the previous studies on numerical taxonomy and multilocus enzymes electrophoresis patterns of the 12 rhizobial strains isolated from Shapotou region, the contents of G + C mol%, DNA-DNA relatedness and 16SrDNA sequence of the representative strain were tested. The DNA G + C content of the members of this group ranged from 56.4 to 62.2. The values of DNA-DNA hybridization within the group were above 70%, and relatendness between representative strains of this group and known rhizobial species was below 66.6%. The full-length of 16S rDNA sequence of representative strain N220 was compared with the type strains of all known rhizobia species and related bacteria by the PHLIP version 3.572c composed a unrooted phylogenetic tree, the strain N220, R. galegea, two unnamed rhizobial strains(SH19312, SH22623) and three A-grobacterium strains constituted a branch in this tree. The similarity values of 16S rDNA sequence between strain N220 and other strains in this branch were above 95%. PMID- 12555412 TI - [Study of HSP70 gene upstream regulation element on expression efficiency of GST gene in M. smegmatis]. AB - Four different expression vectors were constructed by cloning foreign gene which encode Schistosoma japonicum 26 kD antigen (Sj26GST) into Escherichia coli Mycobacteria shuttle plasmid pBCG-2000 and investigated their expression efficiency in mycobacteria smegmatis. The plasmid which contains promoter of human mycobacterial tuberculosis heat shock protein 70 was firstly digested with Nco I and modified with two different ways to lead to two kinds of SD sequences, and ligated with Sj26GST encoding gene. Then, the DNA fragment contained HSP70 promoter and Sj26GST gene was obtained and cloned into E. coil-mycobacteria shuttle plasmid pBCG-2000, and finally four recombinant mycobacterial expression vectors that differenciated in SD sequence, orientation and copy number were selected. The expressed native recombinant Sj26GST(rSj26GST) was soluble and could be observed on SDS-PAGE about at the molecular weight of 26 kD obviously. Analysis with protein density scanning indicated: the expression efficiency that containing double-copy promoter-foreign gene vector was the highest and the expressed protein which was about 1.6 times than others was 28% of total protein of Mycobacteria smegmatis. The cloned direction and SD sequence had no significant effect on expression efficiency. PMID- 12555413 TI - [Expression of soluble 55 kD receptor for human tumor necrosis factor using baculovirus vector]. AB - Total RNA was isolated from HeLa cells and about 530 bp cDNA encoding shTNFR55 was amplified with RT-PCR from the total RNA. The cDNA was cloned into the downstream of polyhedrin gene promoter in transfer vector pAcGP67B, and a recombinant transfer plasmid pAcTNFR was constructed. Insect cell sf9 was co transferred with recombinant plasmid pAcTNFR and baculovirus AcNPV DNA. Recombinant virus AcNPV-TNFR containing shTNFR55 gene were formed by homologous recombination in insect cells. They were purified by plaque assay and were confirmed by DNA dot blot. The biological activity of expressed product was determined with culturing TNF-susceptible L929 cell line, and the result showed that the expressed product could neutralize the cytotoxicity of TNF on L929 cell line. Ligand blot analysis revealed the expressed protein three discrete bands between 20-25 kD. PMID- 12555414 TI - [Preliminary study of P19 gene from Bacillus thuringiensis subsp israelensis]. AB - The P19 gene and cyt1A gene were obtained by PCR with 9.7 kb HindIII fragment containing P19 gene and cyt1A gene from 72 MD plasmid of Bt as a template. Digested PCR products were ligated to expression vector pUHE24 and transformed into E. coli XL-1. Three clones, LZ19 harboring P19 gene, LZcyt1A harboring cyt1A gene, LZ19A harboring P19 gene and cyt1A gene, were screened. The growth cure of cloned strains were determined under IPTG induction. The result showed that pLZ19 did not affect the growth of E. coli, pLZcyt1A is typically Lethal for E. coli, Lethal initial efficiency of pLZ19A is much higher than that of pLZcyt1A. Probably, this was a result that P19 gene enhanced initial expression of cyt1A gene in E. coli. PMID- 12555415 TI - [Species identification of Mycobacterium tuberculosis with insertion elements]. AB - The Insertion Sequence(IS) element IS6110 and IS1081 of M. tuberculosis complex were used as template to prepare probes for typing strains by means of restriction fragment length polymorphisms (RFLP). Chromosomal DNA of M. tuberculosis H37Ra, H37Rv, M. bovis digested with EcoR I, Pst I, Sal I, BamH I, Pvu II was subjected to Southern Blot analysis With labeled 317bp from IS6110 and 248 bp from IS1081. The results indicated that for M. tuberculosis complex, IS1081 cannot be used to type strains, IS6110 can be used to distinguish strains into broad groups and is an excellent epidemiological tool for studying outbreaks. An investigation of forty-six M. tuberculosis isolates by IS6110 RFLP revealed that M. tuberculosis strains not only are of highly polymorphic, but also are presentin multiple copies ranging from 7 to 18 copies. PMID- 12555416 TI - [Comparison of lignocellulolytic enzyme profiles secreted by Panus conchatus and Phanerochaete chrysosporium during solid state cultures]. AB - Most of white-rot fungi, such as Phanerochaete chrysosporium, can cause severe concomitant cellulose degradation during biodegradation of lignocellulose. Panus conchatus, a white-rot fungus, can cause efficient delignification of straw with only limited concomitant cellulose degradation. The results in comparison of lignocellulolytic enzyme profiles secreted by P. conchatus and P. chrysosporium during solid state cultures have shown that laccase and Mn-dependent peroxidase are main lignin-degrading enzymes of these two fungi respectively; high activities of xylanase are secreted by both fungi; and much lower activities of cellulases i.e. endo-glucanase, avicelase and cellobiase, especially endo glucanase, are produced by P. conchatus during the whole cultures. The results further confirm that Panus conchatus has ability of strong selective delignification of lignocellulose. PMID- 12555417 TI - [Purification and characterization of alkaline xylanases from Pseudomonas G6-2]. AB - Pseudomonas G6-2 produced two extracellular xylanases, named XynA and XynB. The enzymes were purified by ammonium sulfate fractionation, Sephadex G-100, DEAE Sephadex, CM-Sephadex and Bio-gel P-10 chromatographies. Both enzymes were indicated to be endoxylanases, which produced oligomers of xylose from xylan and did not hydrolyze it to xylose. They had same temperature optimum(50 degrees C) and different pH optimum(pH 7.0-9.8 for XynA and pH 7.0-8.0 for XynB). At pH 7.6 and 65 degrees C, XynA and XynB possessed the half life of 6 min and 140 min, respectively. Their activities were strongly inhibited by Cu2+, Fe3+, Pb2+, Zn2+ and Hg2+. The results of chemical modification indicated that tryptophan and carboxy group were related to active center. PMID- 12555418 TI - [Studies on the lactate oxidase producing conditions by Edwardsiella tarda]. AB - A bacterium producing lactate oxidase was re-screened from five strains based on previous researches. The lactate oxidase activity was the highest in strain L1 and this stain was chosen as the enzyme source. Morphological and physiological studies revealed that the bacterial strain L1 belongs to the Edwardsiella tarda Biogroup I. This stain is different from the reported strains Mycobacterium and Pediococcus, which produce lactate oxidase. The enzyme producing conditions were studied in shaking cultures, and the effects of initial pH, riboflavin, lactate and ammonia sulphate concentrations on the production were carried out respectively. The bacteria resource of enzyme is significant to pyruvate production by enzymatic method, and to the enzyme assay of lactate for medical diagnosis, and the application of enzyme electronic probe. PMID- 12555419 TI - [Construction of thermotolerant ethanol-producing yeast by protoplast fusion]. AB - Protoplast fusion between Saccharomyces cerevisiae A001 and Kluyveromyces Y034 has been studied. A001 is a high alcohol yield yeast and can ferment maltose into ethanol, but can not grow at 45 degrees C. However, Y034 can grow above 45 degrees C but has a lower alcohol yield and can not ferment maltose. We obtained the fusants which were tested in several aspects such as cell morphology, physiological and biochemical feature, isozyme patterns analysis, alcohol fermentation at high temperature(45 degrees C) and so on. The fusant AY023 with the final ethanol concentration of 7.4% has been selected, which is thermotolerant yeast with the highest alcohol yield at 45 degrees C until now. PMID- 12555420 TI - [Solid-state fermentation with Penicillium sp. PT95 for carotenoid production]. AB - A preliminary study on solid-state fermentation (SSF) with Penicillium sp PT95 for carotenoid production was performed. The results showed that the production of carotenoid in sclerotia of PT95 was more efficient in corn meal medium than in either wheat bran medium or cottonseed hull medium. Addition of nitrogen and carbon sources as well as vegetable oil to media was required for increasing the dry weight of sclerotia and carotenoid yield. Among several tested compounds for nitrogen and carbon sources, sodium nitrate and maltose were the best. Through orthogonal experiments, the optimum culture medium was obtained by supplement of NaNO3 3g, maltose 10 g, soybean oil 2.5 g to per liter of salt solution. Under the optimum culture conditions, the sclerotia dry weight increased from 5.36 g to 9.70 g per 100 g dry substrate, the carotenoid yield from 2149 micrograms to 5260 micrograms per 100 g dry substrate, the proportion of beta-carotene in carotenoids from 61.4% to 71.3%. PMID- 12555421 TI - [The identification of 5 new serotypes of Bacillus thuringiensis from soil in China]. AB - Five new strains of Bacillus thuringiensis(Bt) from soil in China were classified. 5 new H-serotypes: H42, H43, H56, H60 and H62 were identified. There were some distinction in the morphology, biochemical reaction and toxicity against species of insect. Five new Bt subspecies were identified as follows: Strain YGd22-03 Bacillus thuringiensis subsp. jinghongiensis, H42; Strain KK31-01 Bacillus thuringiensis subsp. guiyangiensis, H43; Strain SCg04-02 B. thuringiensis subsp. rongseni, H56; Strain NXP15-04 B. thuringiensis subsp. pingluonsis, H60; Strain HZ39-04 B. thuringiensis subsp. zhaodongensis, H62. PMID- 12555422 TI - [Studies on the parasites of Paris polyphylla var. yunnanensis]. AB - Two bacteria and three fungi were isolated from the hypogeal stems of Paris polyphylla var. yunnanensis. The bacteria were identified as Bacillus cereus and Pseudomonas alcaligenes, and the fungi were identified as Periconia sp., Pachnocybe albida and Hormomyces paridiphilus. The results on liquid culture of B. cereus, P. alcaligenes and H. paridiphilus indicated that the colloidization and polysaccharide content increasing in hypogeal stems of P. polyphylla var. yunnanensis were due to the extracellular polysaccharide secretion of the parasitical fungus H. paridiphilus. PMID- 12555423 TI - [Cloning of glycoprotein B gene from strain Rispens of Marek's disease virus and construction of recombinant fowlpox virus]. AB - Purified DNAs from Chicken Embryo Fibroblast (CEF) cultures infected with MDV strain Rispens were used as templates. Specific fragment with the size of about 2.9 kb was successfully amplified through Polymerase Chain Reaction(PCR) and identified to be gB gene of MDV by dot blot hybridization with a digoxigenin labelled MDV gB specific oligonucleotide probe. The gB gene from strain Rispens was cloned into pUC19 and FPV insertion vector pFG1175-1 to construct plasmid pMGB and pFGBR1775-1 respectively. DOSPER liposome-mediated transfection with insertion vector DNA pFGBR1175-1 was performed on CEF monolayers infected with FPV 3-4 h earlier. Recombinant FPV was clone purified. Immunofluorescence Assay(IFA) showed that MDV gB gene had been expressed in FPV. PMID- 12555424 TI - [Expression of VP7 from a group A rotavirus G4 field strain in baculovirus system]. AB - The full length of G4 type VP7 cDNA was amplified by reverse transcription and polymerase chain reaction (RT-PCR) from a group A rotavirus field strain CR117 circulated in Beijing. The amplified cDNA was first cloned into pTZ18. Sequence analysis showed 96.9% homology of the deduced amino acid between VP7 of CR117 and ST3. Then the VP7 cDNA of CR117 was subcloned into the transfer vector pBacPAK8 downstream of the polyhedrin promotor. Insect cells were cotransfected with recombinant plasmid pBacPAK8-CR117 and Bsu36I-digested BacPAK6 viral DNA. Recombinant baculovirus containing CR117 VP7 gene was obtained by plaque screen and hybridization identification. Specific VP7 with three different molecular weight was detected by Western blot using hyperimmune serum against Rotavirus strain ST3 (G4 type) when expressed in insect cells. PMID- 12555425 TI - [Detection of hypodermal and hematopoietic necrosis baculovirus by PCR method]. AB - For control of the explosive epidemic disease of Penaeid shrimp in China. We have developed a method for detection of Hypodermal and hematopoietic necrosis baculovirus (HHNBV) by polymerase chain reaction (PCR). Two pairs of PCR primers (DP1/DP2 and DP3/DP4) were prepared, based on the RFLP analysis between HHNBV and penaeid rod-shaped DNA virus. PCR using either DP1/DP2 or DP3/DP4 specified HHNBV genome. When PCR products amplified with DP1/DP2 were subjected to nested PCR with DP3/DP4, a high sensitivity about 0.04 pg was seem with the nested PCR. PMID- 12555426 TI - [The effects on ethanol fermentation of Saccharomyces cerevisiae by adding Ca2+ and inositol]. AB - In this paper we report the effects of Ca2+ and inositol upon growth, ethanol fermentation and ethanol tolerance of Saccharomyces cerevisiae. The result shows that by adding inositol has notable effects on growth and ethanol tolerance of Saccharomyces cerevisiae (p < 0.05), but has little effects on ethanol fermentation (p = 0.3933). By adding Ca2+ has remarkable effects on ethanol fermentation and ethanol tolerance (p < 0.05), but has little effect on yeast growth (p = 0.1544). By means of adding Ca2+ and inositol has more effects on the growth, ethanol fermentation and ethanol tolerance than the condition of only adding Ca2+ or inositol separately. PMID- 12555427 TI - [Preparation, purification and identification of sialic acid from Escherichia coli C-8]. AB - Colominic acid produced from Escherichia coli C-8 was purified by ammonia sulfate precipitation, dialysis and concentration. After hydrolysis of colominic acid at a pH of 2.5, at 70 degrees C for 4 h, sialic acid was obtained, then purified by chromatography on Dowex1-x8. Analysis of thin-layer chromatography and absorption spectrum of sialic acid in the orcinol/Fe3+/HCl and the periodic acid/thiobarbituric acid confirmed that its purity was identical with that of standard sample. PMID- 12555428 TI - [Studies on the screening high yield acid protease producing strain L336 by combining microwave irradiation with chemical inducing]. AB - A high yield acid protease producing strain Aspergillus usamii L336 was screened from its parent strain B1 by combining microwave irradiation with inducing of nitrosoguanidine and Li2SO4. By incubation of L336 in shaking flask at 31 degrees C for 88 h, its enzyme activity was 5500 u/mL, and its properties for high yield producing acid protease remained stable after many times of subculture and storage for two months. PMID- 12555429 TI - [Advance of the research on enzyme inhibitors of microbial origin]. PMID- 12555430 TI - [Can the supraventricular proarrhythmic effects of class 1C antiarrhythmic drugs be prevented with the association of beta blockers?]. AB - Due to their electrophysiological characteristics, class 1 antiarrhythmic drugs can induce an auricular flutter with a 1/1 response. In addition to antiarrhythmic treatment, several authors have therefore considered using drugs capable of slowing auriculoventricular nodal conduction and preventing the 1/1 response. Beta-blockers have been proposed as candidate drugs. In this study, two patients were treated with an association of class 1 antiarrhythmic drugs (cibenzoline in one case, flecainide in the other) and beta-blockers. The administration of these drugs resulted in an atrial proarrhythmic response, and wide QRS tachycardia. Although both subjects had underlying heart disease, the tachycardia was relatively well tolerated in both instances. It was concluded that although beta-blockers may not suppress the risk of atrial proarrhythmia, they at least permit an improved tolerance to this complication. PMID- 12555431 TI - [Angor with occlusion of the left main coronary artery treated with angioplasty]. AB - The occlusion of the left main coronary artery is rare and generally fatal. However, some subjects do survive. The two conditions necessary for survival appear to be the existence of a dominant right coronary artery and above all a rapidly functional left-right collaterality. The time lapse between occlusion and the introduction of an efficient collateral system is fundamental to patient survival. If the time lapse is too long, there is a risk of myocardial infarction, often complicated by cardiogenic shock, with a poor prognosis even after deocclusion. In this study, the case is reported of a patient with left main coronary artery occlusion presenting as unstable angor without an increase in enzymatic levels, and with a totally functional left-right collaterality. Deocclusion angioplasty was successfully performed. Finally, the short- and medium-term results of left main coronary artery angioplasty have been discussed. PMID- 12555432 TI - [Myocardial pseudo-infarction: "stress"-associated catecholamine-induced acute cardiomyopathy or coronary spasm?]. AB - As well as pheochromocytoma, in which it has been established that an excess of circulating catecholamines is responsible for the development of catecholamine induced acute myopathy, some rare cases have been reported of a similar cardiac incident following intense emotional stress. In this study, the case has been examined of a 56-year old female with no history of cardiovascular disorder who presented with intense, nitro-resistant prolonged chest pain mimicking an acute coronary syndrome immediately following a situation involving major psychological stress. The admission electrocardiogram revealed a sharp decrease in R-wave amplitude in the right chest leads associated with an extended QT interval, and secondarily with subepicardiac ischemia in the lower leads. However, a few days after admission the electrical signs and septo-apical akinesia that had initially been observed by echocardiography completely disappeared. The clinical examination ruled out a diagnosis of myocardial necrosis, acute myocarditis, or pheochromocytoma. Moreover, no direct evidence of coronary spasm was found. The outcome was positive, with complete reversibility of all clinical signs and no organic sequelae. It is considered that this was probably a case of catecholaminergic acute cardiomyopathy triggered by intense emotional stress, a rare occurrence that should nevertheless be systematically taken into account in cases with similar clinical signs. PMID- 12555433 TI - [Early postoperative constrictive pericarditis: value of echocardiography. Report of 4 cases]. AB - In the present study, four cases of early postoperative constrictive pericarditis have been described which serve as a basis for recalling the current main echographic features of this disorder: pericardial thickening, abnormal septal movement with inspiratory expansion of the right ventricle, respiratory variations in ventricular filling, characteristic modifications in the supra hepatic pulmonary venous flow, and pulmonary insufficiency. Some hypotheses have also been presented on the possible etiology of this disease. The necessity of making a precise and rapid diagnosis, which should also be confirmed by catheterization to ensure the appropriateness of therapy, has been emphasized. Pericardectomy can determine the long-term prognosis. PMID- 12555434 TI - [Three-dimensional echocardiography: methodology trial]. AB - A method has been developed for systematic and reproducible exploration by three dimensional echocardiography. An associated technical terminology has also been introduced, and the application principles have been examined together with a number of useful clinical examples. After a period of technical and clinical validation, three-dimensional echocardiography has now entered the stage of practical clinical application, in particular for mitral valve disorders and intraauricular communications. The methodology that has been proposed is intended to provide user training, facilitate communication with teams working in the field, and also between echography operators and surgeons, and catheter or clinical staff. PMID- 12555435 TI - [Alcohol and rhythm disorders]. AB - Cardiovascular disease is the main cause of mortality in chronic alcoholics. There is a clear association between excessive alcohol consumption and the risk of sudden cardiac death. The pro-arrhythmogenic effect of ethanol could be responsible for some of these cases of arrhythmia and sudden death in subjects with an alcoholic cardiomyopathy and also in those with an apparently normal heart. In any case of supraventricular or ventricular arrhythmia in a chronic alcoholic or in an occasional heavy drinker, the potential role of alcohol consumption in the initiation of these disorders should be considered. In all cases, patient management consists of detoxification and abstaining from alcohol consumption, but the withdrawal period is particularly critical as regards the risk of ventricular arrhythmias. PMID- 12555436 TI - [Hemostasis disturbances in myocardial ischemia]. AB - In this article, a description has been given of the close connection between coronary atherosclerosis and the onset of thrombosis. The hemostatic factors examined in this study are implicated both in the pathology of acute coronary syndromes and in the prognosis of ischemic heart disease. Amongst other factors, the role of the following has been investigated: platelets, thromboxane A2 and prostacyclin, von Willebrand factor, factor VII and tissue factor, thrombin, fibrinogen tissue plasminogen activator and plasminogen activator inhibitor. It is concluded that endothelial dysfunction in coronary atherosclerosis is the most frequent cause of disturbances in hemostatic function. PMID- 12555437 TI - [The art of medicine and the art of leadership]. PMID- 12555438 TI - [Schizophrenia--a mystery]. PMID- 12555439 TI - [Does infection prevention prevent infection?]. PMID- 12555440 TI - [Hepatitis C in Northern Norway--an 8-year material]. AB - BACKGROUND: In 1992 a screening and medical follow-up programme of patients with hepatitis C was established in northern Norway. We report the data from the eight year observation period. MATERIAL AND METHODS: Patients with positive recombinant immunoblot assay (RIBA) confirmation tests of hepatitis C from the counties of Nordland, Troms and Finnmark were referred from primary care. RESULTS: At the end of 1999, 1,132 RIBA positive hepatitis C patients were registered, giving an estimated prevalence of 0.24%. 236 patients were included in the follow-up study. 75% were between 17 and 39 years of age. Drug abuse was the mode of transmission in 67%. Mean age by the time of transmission was 23 years (range 12-63 years). 103 patients were treated; 136 patients did not fulfill the criteria for treatment. At the time of inclusion, 76% had no clinical symptoms. 36% treated with monotherapy and 47% treated with combination therapy had no detectable virus six months after end of treatment. INTERPRETATION: Hepatitis C infection is a disease among young people; intravenous drug abuse is the most frequent mode of transmission. The initial phase of the disease has few symptoms. A hepatitis C programme run by public health authorities is highly recommended. PMID- 12555441 TI - [Vertical transmission of hepatitis C virus in Sor-Trondelag]. AB - BACKGROUND: The aim of this study was to investigate the vertical transmission rate of hepatitis C virus (HCV) in a Norwegian county and describe the clinical outcome of the infected infants. MATERIAL AND METHODS: Over the 1991-2000 period, 29,433 pregnant women gave birth to 37,893 children. Women born in high endemic areas and those with a history of drug abuse were offered anti-HCV testing. All anti-HCV positive specimens were subjected to supplementary testing by RIBA and PCR. RESULTS: Hepatitis C virus antibodies were detected in 61 women who delivered 83 children over this ten-year period. 61% of the women were HCV-RNA positive. The transmission rate was 5.0% (8.3% for HCV-RNA positive mothers). None of the HCV-RNA negative mothers transmitted the virus to their offspring. 95% of the anti-HCV positive mothers had a history of drug abuse; none came from high endemic areas. All mothers tested negative for HIV and hepatitis B virus (HBs antigen) during pregnancy. HCV antibodies persisted up to 18 months after birth in non-infected infants, but most of them tested negatively within one year of age. All infected infants tested negative by PCR at birth, and only one out of four had persistently elevated alanine amino-transferase. INTERPRETATION: Our data is consistent with other studies of vertical transmission of HCV. Anti-HCV screening of larger groups of pregnant women is warranted to investigate the true incidence of vertical transmission in our population. PMID- 12555442 TI - [Hepatitis B in children--diagnosis, follow-up and treatment]. AB - 350 million people worldwide are chronic carriers of the hepatitis B virus. Mainly because of immigration, the number of children with chronic hepatitis B infection in Norway is also increasing, although the absolute number is still small. In April 2001, a group of Norwegian paediatricians interested in infectious diseases held a meeting to discuss the clinical management of chronic hepatitis B in children and develop recommendations. The recommendations are based on current European and American guidelines, experience from England and Sweden, and a review of the literature. International epidemiological data and data from Norway are briefly presented, followed by recommendations regarding diagnosis, follow-up and treatment of chronic hepatitis B infection in children. Children at risk of contracting hepatitis B from their mothers should be immunized shortly after birth. Paediatricians should follow up children with chronic hepatitis B infections in order to identify those who may be eligible for treatment. PMID- 12555443 TI - [Hepatitis C in children--diagnosis, follow-up and treatment]. AB - Hepatitis C-virus infection is relatively infrequent among children in the western world. Although most hepatitis C infections in children evolve to chronicity, liver damage is usually mild. In April 2001, a selected group of Norwegian paediatricians interested in infectious diseases convened to discuss the clinical management of hepatitis C in children. So far, no consensus reports concerning follow-up of hepatitis C infected mothers and their children or clinical management of chronic hepatitis C infection in childhood have been published. In view of the limited experience with hepatitis C among Norwegian children, strategies for the clinical management of hepatitis C infection are discussed based upon available literature and experience from England and Sweden. We present epidemiological data, the risk of vertical transmission and the clinical characteristics of hepatitis C in children. Procedures for follow-up of hepatitis C infected mothers and their children and guidelines for treatment of hepatitis C infected children are proposed. Long term follow-up to identify those who require treatment is important. PMID- 12555444 TI - [Antenatal care in Norway--many unnecessary check ups]. AB - BACKGROUND: Until recently antenatal care in Norway has been provided solely by general practitioners. In 1995, it was laid down in law that the communities should offer antenatal care provided by midwives in community health centers. The resulting conflict between midwives and disagreeing general practitioners may have led to an increase in the number of antenatal visits. Also, the utilization of midwife-based antenatal care is unknown. MATERIAL AND METHODS: National cross sectional study including all 54 hospital departments of obstetrics. For all patients number of antenatal visits and parity were recorded. The study included 1,780 women giving birth during the two-week registration period. RESULTS: The mean number of antenatal care visits was 12.0 (range 0-44). The difference between primiparous (mean 12.4) and parous women (mean 11.7) was minor. Midwives provided 44% and doctors 56% of the antenatal visits. A total of 279 women (16%) had not seen a midwife during pregnancy; 3% had only seen a midwife and no doctor. INTERPRETATION: The recommended reduction in the frequency of antenatal visits is not followed up. The proportion of visits performed by midwives is approaching the 50% level suggested in the guidelines for antenatal care. PMID- 12555445 TI - [Ultrasonography in primary health care--experiences within obstetrics 1983-99]. AB - BACKGROUND: In 1983 ultrasound for obstetric purposes was introduced in a rural general practice in south-eastern Norway. We wanted to evaluate its systematic use in terms of pregnancy management and birth outcome. MATERIAL AND METHODS: Four sets of data were analysed: All obstetric ultrasound examinations done at the Sel Municipality Health Centre from 1 January 1998 to 28 February 1999; the database at the county hospital covering all obstetric patients and births in its catchment area, including those from the Sel community; all Medical Birth Registry reports on women from Sel from 1983 to 2000; a local register of pregnancy management and events in Sel from 1983 to 2000, organised by the local general practitioner. RESULTS: We found no difference in the accuracy of assessment of date of birth between the general practitioner in Sel and the Lillehammer outpatient obstetric department. The average number of ultrasound scans of individual women was also the same. Obstetric ultrasound in general practice successfully selected out patients who could be managed locally and those who needed referral. INTERPRETATION: Our results indicate true benefits and no definite hazards of obstetric ultrasound in rural general practice. As we have evaluated only one single practice, these results should be confirmed or negated in future studies. PMID- 12555446 TI - [The founder of the first therapeutic community in Norwegian psychiatry]. AB - In 1959, Dr. Herluf Thomstad (1905-2000) started the first therapeutic community in Norway at Ulleval University Hospital. Meetings in the community brought together all patients and staff members. Patient activities included catering and cleaning, but also the important tasks of informing and taking care of new patients. At a later point in time they were also entrusted with chairing the community meetings to which reports were presented from the activities the day before. The staff discarded their uniforms. Nurses led the patients in their daily duties and thus were coworkers in the development of modern milieu therapy. Thomstad supervised nurses and physicians alike: individual therapy should go hand in hand with milieu therapy in order to enhance the benefit to patients. During the first years, Dr. Thomstad met with a great deal of resistance from fellow Norwegian psychiatrists, but he never gave up. In 1971 he received the order of St. Olav, first class. His assistants developed new methods of family psychiatry; many of them became chairs of other departments of psychiatry in Norway. PMID- 12555447 TI - [Interactions with antiepileptics]. PMID- 12555448 TI - [Schizophrenia--incidence and significance]. AB - The human suffering associated with schizophrenia is enormous--for patients, families, and in society at large. 10% of all disabled citizens in Norway have schizophrenia, although the yearly incidence is low (about 7-15 per 100,000). This variance in incidence is found in most countries. There is an increased risk for schizophrenia among the urban born compared with those born in rural areas, a declining incidence over time, and increased risk in offspring of migrants. In Norway, the total cost per year for schizophrenia is estimated at 4 billion NOK (1995). In western countries costs are estimated to exceed 1% of the national budget. There is no other disorder with comparable costs. Schizophrenia alone costs more than all types of cancer and more than all cardiovascular diseases. There are reasons to believe that not all patients with schizophrenia are given treatment that is in accordance with good clinical practice. The time lag from on set of manifest psychosis and to appropriate treatment is often long--in most western countries up to 2-3 years. Early detection and treatment of schizophrenia is an important challenge. PMID- 12555449 TI - [Early determination of schizophrenia--is primary prophylaxis possible?]. AB - BACKGROUND: Early intervention is a new and potentially promising strategy for the treatment of schizophrenia. MATERIAL AND METHODS: The article reviews the recent literature on how primary prevention can be conceptualised and what strategies may be effective. RESULTS AND INTERPRETATION: Primary prevention of schizophrenia means that treatment is given during the prodromal or prepsychotic phase. The DSM-III-R criteria for prodromal symptoms are too unspecific to be used to target such prevention. The basic symptom model seems to give the best concepts for primary prevention. Three types of prepsychosis are defined: attenuated positive symptoms, brief intermittent psychotic symptoms, and genetic risk with recent psychosocial deterioration. Preliminary findings indicate that 33-70% of those being diagnosed with prepsychosis develop psychosis within the first year of follow-up. Psychiatric health services should be organised in such a way that patients are given treatment as soon as possible after the onset of psychosis. PMID- 12555450 TI - [Neurocognitive deficits in schizophrenia]. AB - BACKGROUND: Patients with schizophrenia experience difficulties in a broad spectrum of neurocognitive domains. MATERIAL AND METHODS: A review of neuropsychological findings in schizophrenia and of cognitive remediation studies in schizophrenia is provided, based on relevant reports identified by a survey of the literature. RESULTS AND INTERPRETATION: The most pronounced neurocognitive deficits in patients with schizophrenia are in the areas of attention, memory and executive functions. There is some empirical evidence to the effect that specific cognitive functions can be improved through cognitive training programmes. There is, however, no evidence indicating that improved performance on specific neuropsychological tasks can be generalized to other tasks or areas. PMID- 12555451 TI - [Models and trends in psychiatry--should neurobiology form a common basis?]. PMID- 12555452 TI - [American guidelines for treatment of borderline personality disorder]. PMID- 12555453 TI - [Hand weapons and health]. PMID- 12555454 TI - [Don't forget antacids!]. PMID- 12555455 TI - [Wrong information concerning rofecoxib]. PMID- 12555456 TI - [Health plans and health insurance in American health care]. PMID- 12555457 TI - [With a computer in the stomach]. PMID- 12555458 TI - [Progesterone cream]. PMID- 12555459 TI - [Cultural surgery]. PMID- 12555460 TI - [Euthanasia in the Netherlands--new, alarming statistics]. PMID- 12555461 TI - [Comparative study of post-infarction myocardial viability after fibrinolysis by stress tomoscintigraphy and echography: can viability be detected without ischemia?]. AB - The objectives of this prospective study was to define the comparative ability of stress myocardial scintigraphy and dobutamine stress echocardiography to demonstrate post-MI myocardial viability, assessed on the functional recovery in terms of improvement of global and segmental kinetics by cardiac gamma angiography after revascularization. 18 patients (11 anterior MI, 7 lateral or inferior MI) and 162 segments were analysed semiquantitatively. All patients with persistent significant stenosis underwent secondary revascularization of the artery responsible for myocardial infarction. The prevalence of viability was high, as only 34% of segments initially presented a segmental kinetic abnormality and contraction was improved at 6 months in 54% of cases. Stress scintigraphy and dobutamine echocardiography detected viability with a sensitivity of 96% and 70%, a specificity of 88% and 82%, a positive predictive value of 89% and 77% and a negative predictive value of 95% and 76%, respectively. Only the wall score index with low-dose dobutamine was correlated with the ejection fraction at 6 months. Stress echocardiography is a more reliable predictor of the degree of functional recovery after revascularization. Scintigraphy visualizes much more extensive abnormalities than echocardiography. This often corresponds to ischaemic territories with normal contraction under baseline conditions and low doses of dobutamine. It therefore seems preferable both examinations for optimal assessment of thrombolized patients following myocardial infarction. PMID- 12555462 TI - [Predictive factors of atrial fibrillation in mitral stenosis. Clinical and echocardiographic study]. AB - The objective of this study was to define the predictive factors of atrial fibrillation in pure or very predominant mitral stenosis in a series of 472 consecutive patients divided into 2 groups according to the presence (group I: n = 113) or absence (group II: n = 359) of permanent atrial fibrillation. Univariate analysis showed that predictive factors for atrial fibrillation in mitral stenosis are age (40.3 +/- 9 years vs 31.4 +/- 9.5, p < 0.0001), history of commissurotomy or mitral angioplasty (13 cases vs 10, p < 0.01), functional class III or IV (36 cases vs 43, p < 0.01), history of valvular heart disease (8.4 +/- 7.3 years vs 6.4 +/- 9.2, p < 0.05), left atrial diameter (53.3 +/- 10.3 mm vs 46.5 +/- 8.5, p < 0.0001) and mitral surface area (1.1 +/- 0.4 cm2 vs 1.3 +/- 0.4, p < 0.0001). On multivariate analysis, age and left atrial dilatation were independent predictive factors of atrial fibrillation in mitral stenosis. PMID- 12555463 TI - [Multiple cholesterol emboli syndrome: beneficial effects of early heparin therapy. A case report]. AB - The multiple cholesterol emboli syndrome (MCES) is a rare, multi-organ disease than can occur spontaneously or after arterial or cardiac catheterization, arteriography, angioplasty, cardiovascular surgery, oral or intravenous anticoagulation, systemic fibrinolysis and cardiorespiratory resuscitation, predominantly in male subjects with disseminated atherosclerosis over the age of 60 years. Clinical signs of MCES vary considerably depending on the organs involved, but the signs most frequently encountered are renal failure, skin lesions (livedo reticularis, purple toc, ulcers, etc) and transient eosinophilia. Optimal treatment of this syndrome is controversial and is often symptomatic. However, the most effective measure remains prevention based on identification of high-risk patients, treatment with platelet antiaggregants and careful handling of catheters. This syndrome has a serious prognosis in the majority of cases. In this article, the authors describe a case of MCES. After thoracic aortography, this 73-years-old patient presented typical clinical sign of MCES (angina, cerebrovascular accident, bilateral blindness, transient renal failure and splenic infarction). The clinical course was favourable in response to heparin therapy and splenectomy and caudal pancreatectomy. Histology confirmed the presence of cholesterol emboli in the lumen of splenic arterioles. Except in the case of severe bleeding diathesis, the authors recommend early heparin therapy for MCES caused by catheterization, angioplasty or cardiovascular surgery. However, complementary studies must be performed to more clearly define the effects of heparin on MCES. PMID- 12555464 TI - [Right atrial hemangioma presenting as pericardial tamponade. A case report]. AB - Intracardiac haemangioma is a very rare benign primary cardiac tumour. A 40-year old patient, with no notable history, presented with chest pain, accentuated by deep inspiration, associated with NYHA stage III dyspnoea. Physical examination revealed signs of right heart failure and electrocardiogram showed low voltage with diffuse repolarization disorders. Chez x-ray showed a cardiothoracic index CTI > 0.70, and enlarged right margins. A large pericardial effusion with a tumour mass filling the right atrial cavity was discovered on transthoracic echocardiography. Chest computed tomography demonstrated the tissue origin of the tumour and suggested extension to the inferior vena cava and hepatic veins. The patient was referred to a surgeon for tumour resection. Histological examination revealed intracardiac capillary haemangioma. Five month postoperative follow-up did not reveal any abnormalities. PMID- 12555465 TI - [Ischemic pseudo-normalization of T waves and ST segment]. AB - The development of acute myocardial ischaemia in a territory with baseline repolarization abnormalities can be reflected by transient pseudo-normalisation of the T wave and/or ST segment. These repolarization abnormalities can occur spontaneously, during a stress test or during of an isoproterenol test. Clinicians should be familiar with these unusual electrocardiographic changes of acute ischaemia, which require appropriate surveillance and treatment. PMID- 12555466 TI - [Role of tilt test in the diagnostic and therapeutic management of syncopes]. AB - Following confirmation of one or several episodes of syncope, the most important step is identification of the cause. Unfortunately, despite a careful actiological work-up, a cause is not found in about 30 to 40% of cases, but the tilt test can attribute some of these cases to a vagal cause. The test must be performed according to a rigorous methodology and interpretation must be based on a good knowledge of abnormalities of the autonomic nervous system, particularly the pathophysiology of vagal syncope. The tilt-test can also be useful to evaluate the efficacy of drug treatment prescribed to prevent recurrent syncope. PMID- 12555467 TI - [Physiopathology of acute coronary syndromes]. AB - Coronary atherosclerosis and its thrombotic complications represent one of the leading causes of lesions usually consists of successive acute episodes, either silent or in the form of an acute coronary syndrome such as unstable angina, non Q-wave myocardial infarctions, transmural myocardial infarctions or sudden death. This mode of progression does not exclude phases of regression, or more frequently stabilization of plaques, which, depending on their haemodynamic repercussions, are then responsible for chronic myocardial ischaemia. Acute coronary syndrome (ACS) correspond to the same pathophysiological process: rupture of an atheromatous plaque initiating harmful thrombotic, inflammatory and vasomotor phenomena. This is not a new concept, but progress over recent years suggests that the composition and biology of the plaque are factors involved more in the initiation of ACS than the size of the plaque. "Soft" lesions, rich in lipids, are clearly not only the most unstable lesions, but also the most thrombogenic because of their large tissue factor content. After describing the structure of vulnerable plaques, the authors discuss the causes of their rupture and the resulting cascade or events, responsible for life-threatening clinical situations. PMID- 12555469 TI - [Do we need a new strategy in hypertension treatment?]. PMID- 12555468 TI - [How to handle cardiovascular treatments during general anesthesia?]. AB - The risk of interference with anaesthesia and the risk of decompensation of the disease must be considered for each drug. Beta-blockers reduce the body's capacity to react to hypovolaemia, but they are beneficial by limiting the response to nociceptive stimuli and by reducing the incidence of myocardial ischaemia. Treatment should therefore be continued. Interferences with the dihydropyridine class of calcium channel blockers on peripheral vasomotor activity are moderate and additive. These agents are well tolerated and can be continued until the operation. Verapamil and diltiazem have a chronotropic and negative inotropic effect which is additive with that of anaesthetics. Administration of these drugs before the operation does not raise any major problems, nor does their discontinuation. Angiotensin-converting enzyme (ACE) inhibitors, used in hypertension and heart failure, considerably reduce the tolerance of factors modifying the blood pressure equilibrium, especially hypovolaemia or haemorrhage. In practice, ACE inhibitors carry a risk of intraoperative hypotension, requiring the use of vasopressor amines to maintain blood pressure. Continuation of treatment has only minimal advantages, regardless of the indication and there is no risk of rebound effect after stopping treatment. The treatment washout period depends on the duration of action of the product. However, discontinuation of treatment is never imperative, regardless of the context. PMID- 12555470 TI - [Role of microcirculation in hypertension]. AB - A large share of the pressure gradient in the vascular system is situated in the microcirculation. Most of the mechanisms resulting in an increase of peripheral resistances during hypertension are also located in this network. Experimental studies have shown that the functional characteristics of arterioles are profoundly modified in most hypertension models. The sensitivity to vasoconstrictor agonists is increased. The activity of the angiotensin-converting enzyme, situated in the arteriolar wall, is considerably greater in genetically hypertensive animals than in control animals. Endothelium-dependent vasodilator mechanisms are also decreased in these animals, while endothelium-derived vasoconstrictor factors are responsible for an increased smooth muscle response. These functional disorders are associated with structural modifications of the microvascular network. Changes of wall thickness are observed only inconstantly, but an initially functional then anatomical rarefaction of arterioles and capillaries has been observed in most experimental models of hypertension and in several clinical studies. This phenomenon reduces the capacities of adaptation of perfusion condition to changes in metabolic requirements of the various organs, reflecting the fact that the microcirculation is a target of hypertensive disease. Antihypertensive drugs should therefore be assessed not only in terms of the reduction of blood pressure figures, but also in terms of their capacity to prevent or correct structural changes of the microvascular network. PMID- 12555471 TI - [Vascular endothelium: target organ of cardiovascular pathologies]. AB - The endothelium plays an essential role in regulation of vascular tone. It has vasodilator, antiaggregating, anti-thrombotic and anti-inflammatory properties. In the case of endothelial dysfunction (induced by hypercholesterolaemia, hypertension and/or diabetes), all of these properties are reversed; it becomes thrombogenic, it secretes vasoconstrictor substances promoting platelet aggregation and penetration of white blood cells into the vessel wall. This endothelial dysfunction then allows the development of the atheromatous lesion. The therapeutic approach must be based on restoration of the normal physiological properties of the endothelium. PMID- 12555472 TI - [Use of carvedilol in daily practice]. AB - Studies conducted over recent years have definitively confirmed beta-blockers as the major treatment for heart failure. However, they are difficult to use and, to date, only carvedilol has been granted a Marketing Authorisation for this indication. The practical aspects of treatment with carvedilol in these patients are reviewed. PMID- 12555473 TI - [Management of aspirin discontinuation in stable coronary heart disease prior to elective surgery]. PMID- 12555474 TI - [Do anti-lipemic agents only work by their lipid-lowering effect?]. PMID- 12555475 TI - [Cardiothyrotoxicosis in the young adult in Basedow disease: report of 30 cases]. AB - The authors report on 30 cases of cardiothyrotoxicosis in the young adult with Basedow's disease. Cardiac arrhythmia represented the most frequent clinical form of dysfunction, mainly atrial fibrillation. Conductive disorders came second, with seven cases of first-degree atrioventricular block [AVB], one case of second degree AVB, and two cases of sinusoidal bradycardia. Three cases of ballooning of the mitral valve were detected by echocardiography. Myocardial hypertrophy was found in one case. The authors discuss the various physiopathological hypotheses regarding conduction and myocardial hypertrophy anomalies. No cases of severe cardiac insufficiency or coronaropathy were noted, which is explained by the absence of cardiopathic antecedents and the young age of the patient population. Treatment is more complicated in the case of a preexisting cardiac event. PMID- 12555476 TI - [Paraganglioma associated with renal artery stenosis: unusual cause of secondary arterial hypertension. A case report and review of the literature]. AB - Secreting paraganglioma is a rare cause of secondary HT and corresponds to an extra-adrenal phaeochromocytoma. This case reports the consecutive coexistence, with a free interval of 10 years, of dysplastic renovascular HT and abdominal paranganglioma in the organ of Zuckerkandl. This is a rare association which requires investigation of a common genetic predisposition (neural crest disease) and emphasizes the importance of long-term surveillance of these patients. PMID- 12555477 TI - [Acute dissection of the ascending aorta complicating aortic coarctation successfully treated with surgery]. AB - Vascular complications and arterial hypertension are major risk factors in the evolution of disease connected with coarctation of the aorta. Supra- and sub stenotic aortic aneurysms are classical findings, whereas a dissecting aneurysm is a rare occurrence. In this study, we report on a case of acute dissection of the ascending aorta complicating a poorly followed-up coarctation of the aorta in a ten-year-old girl. The diagnosis, determined by X-ray of the thorax, was confirmed by echocardiography and magnetic resonance imaging. The first operative stage consisted of treating the aneurysm. After resection of the latter, an impregnated dacron prosthesis was inserted in termino-terminal. The immediate result is satisfactory, and a second operation is planned for the aortic coarctation. PMID- 12555478 TI - [Papillary fibroelastoma of the large mitral valve disclosed by ischemic cerebrovascular accident: a case report]. AB - Thirty-four observations of papillary fibroelastoma (PFE) of the mitral valve have to date been reported in the literature. Our findings are the tenth so far documented on PFE revealed by a cerebral ischaemic vascular accident. The echocardiographic investigation detected a pedunculated tumour of the mitral valve 9 mm in diameter, and associated with moderate mitral leakage. The tumour was surgically excised tumour confirmed the original PFE diagnosis. The evolution seven years after surgery is favourable, with no signs of tumour recurrence and good functioning of the mitral valve system. PMID- 12555479 TI - [Acute aortic insufficiency]. AB - Acute aortic insufficiency can now be diagnosed rapidly and accurately thanks to Doppler echocardiography. The etiologies include infectious endocarditis, aortic dissection, bioprosthesis degeneration and thoracic injury. The clinical diagnosis is substantiated by the particular etiological context, dyspnea and pulmonary edema being the main factors involved. Examination includes finding out whether there is a reduction in the first sound, S1 a generally brief apical diastolic murmur. Echocardiography detects the presence of aortic leakage, the acute character of which is confirmed by the findings of a premature closure of the mitral valve, the existence of telediastolic mitral leakage, a restriction in the transmitral flow, and finally, the absence of left ventricular dilatation. An emergency operation is recommended by most authors in the case of acute aortic leakage due to the major risk or mortality resulting from pulmonary edema, ventricular arrhythmias, electromechanical dissociation or cardiogenic shock. PMID- 12555480 TI - [Behavior of 20 pacemakers as patients pass through 2 models of theft-detection doors]. AB - Episodic interference between the magnetic field of anti thief devices in stores (Electronic Article Surveillance, EAS) and pacemaker function have been reported, but no systematic in vivo study. 20 implanted pacemakers from ten different companies, 1 VVIR, 1 VDD and 18 DDD, were tested with two different EAS systems. #1 uses a 7.5 and 5 kHz field of 670 microTesla and #2 a 72.3 Hz magnetic field, of 1.10 microTesla. Under continuous ECG monitoring, patients had to walk through, and then stay between the doors while moving, and finally to lay against each wall. This was done with their usual pacemaker setting, and then set at their highest sensitivity in the unipolar atrial and ventricular mode. No abnormal PM behavior was observed with standard settings (V sensitivity 1.4 to 5 mV, A sensitivity 0.25 to 1 mV, bipolar (14) or unipolar (6). When programmed at the highest sensitivity, the magnetic field was detected by 5 models, but only with ventricular sensing set at less than 1.2 mV. moreover this only occurred in #2 EAS and not in #1. This resulted in short inhibitions < 1.2 s, and in one safety zone pacing. All occurred in devices conceived before 1993 (6/14) and none after (0/6). EAS devices should not inhibit implanted PM with standard settings, especially in recent models. Ventricular sensitivity should not be programmed in the unipolar mode at 1.2 mV or less. PMID- 12555481 TI - [Heart rate and mortality: review of the literature]. AB - A great number of studies focussed on a wide variety of subjects have been concerned with the subject of morbi-mortality and the effect of heart rate. These studies show that there is a connection between high heart rate and arterial hypertension, and a significant coronary, cardiovascular, non-cardiovascular and overall mortality risk. Two explanations are possible for this. The increase in heart rate together with associated sympathetic hyperactivity can be considered a cardiovascular risk factor: or it could be viewed as an independent risk factor, associated with an increase in mechanical constraints. PMID- 12555482 TI - [Chronotropic incompetence: when should a pacemaker be considered? Which pacing system and which settings?]. AB - Chronotropic incompetence is defined as the inability to increase and maintain heart rate appropriately during exercise. Intolerance to exertion is manifested by a number of clinical symptoms, and is almost obligatory if heart rate cannot be increased. Several rate-modulating pacing systems have been developed. The most obvious and reliable way to increase heart rate during exercise is to detect the sinus node. Adding an atrial lead in a patient in complete AV block and VVI pacing is the most satisfactory way to correct chronotropic incompetence in some patients. Rate-adaptive sensors include motion sensors, respiration sensors, QT interval, and right ventricular contractility. Indications for rate-adaptive pacing should be based on clinical symptoms, demonstration of a lack of cardiac acceleration during exercise, and the presence of another indication for pacing. PMID- 12555483 TI - [Pacemaker syndromes]. AB - The pacemaker syndrome refers to symptoms and signs in pacemaker patients caused by inadequate timing of atrial and ventricular contractions. The lack of normal atrioventricular synchrony may result in decreased cardiac output and increased in atrial pressure which elicits a systemic hypotensive reflex response. These hemodynamic disturbances mostly occur in the presence of retrograde VA conduction is present. The incidence of pacemaker syndrome is uncertain, and varies from 7% to 20% of all ventricular paced patients. Neurologic symptoms or symptoms suggesting low cardiac output or congestive heart failure are indicative of the pacemaker syndrome. These symptoms may vary from mild to severe, they are nonspecific and very common among cardiac patients with or without pacemaker. In many patients they are not even noted until AV synchrony is restored. The pacemaker syndrome is not restricted to the VVI stimulation mode. It can be seen, though rarely, in atrial and even dual-chamber pacing (VDD, DDI, DDD). In these cases it is usually occasional and may be due to inappropriate programming or selection of the pacing mode. PMID- 12555485 TI - [Effect of external electrical interference on pacemakers]. AB - Cardiac pacemaker are sensitive to many kind of electromagnetic interference (EMI). However the low rate of reported effects is due to many protective features. Risks are minimal in everyday life conditions as airport detectors, electronic anti-thief systems, domestic electrical appliances and even mobile phones with the most recent models. Interference detection can be prevented by "medically correct" setting, with the use of bipolar sensing, and ventricular sensitivity above 2 mV. The effect of EMI from the industrial world are difficult to assess and need an individual study. The most important risks occur in the medical world, with the electrocutary, thransthoracic DC shocks, and magnetic resonance imaging. PMID- 12555484 TI - [Infections secondary to the implantation of a pacemaker: update]. AB - Pacemaker lead infection is a major complication of endovascular permanent pacing. The incidence is less than 1% but it is a frequent disease due to the high number of pacemaker implanted. The diagnosis is difficult due to the insidious symptoms. Pacemaker infection must be systematically considered in patients with a pacemaker and symptoms of infection. Several investigations are useful for the diagnosis particularly the transesophageal echocardiography, but all investigations have a low negative predictive value. All of the implanted material must be completely removed. PMID- 12555486 TI - [What methods are available to assess quality of life of patients fitted with a pacemaker?]. AB - Quality of life (QOL) measurements are one of the main determinants for indication and technologic choice in permanent cardiac pacing. QOL measurements are obtained using questionnaire application. Questionnaire for QOL has to be reliable, comprehensive, sensitive and specific. Pacing mode and pacemakers algorithms have been evaluated by QOL measurements: VVI vs DDD, DDD vs VVIR, AAI vs DDD, fall back, algorithm.... Survival data and costs are also relevant aspects of cardiac pacing evaluation. PMID- 12555487 TI - [New pacemaker functions: which ones represent real progress and which are only gadgets?]. AB - Modern pacemakers, and particularly dual chamber models, include a broad array of therapeutic and diagnostic features. Some are designed to increase safety, whereas others are either designed to avoid unnecessary pacing, reduce the current drain and increase the pacemaker longevity, or to improve the hemodynamics. Diagnostic features are more elaborate, more or less automatic, and easier to extract from the pacemaker memory. Some of these new algorithms represent a real advantage, while others appear to be more like gadgets than real advances. Several new algorithms should be improved in order to avoid possibly dangerous side effects. Most of these new features are of little value in the absence of postoperative programming by a well trained physician. PMID- 12555488 TI - [Stimulation in heart insufficiency]. PMID- 12555489 TI - [Monitoring of stimulation probes]. PMID- 12555490 TI - [Paroxysmal junctional tachycardia and normal ECG in sinus rhythm. Is their mechanism influenced by the age of the patient at the time of the study?]. AB - At the time of the curative treatment of paroxysmal junctional tachycardias, to determine the mechanism of the tachycardia is important. Five-hundred forty intracardiac or esophageal electrophysiological studies were performed in patients aged from 9 to 86 years and then revieved. There was no preexcitation syndrome on surface ECG. Programmed atrial stimulation was performed in basal state and if necessary repeated after infusion of isoproterenol. Atrioventricular nodal reentrant tachycardia was the most frequent cause of junctional tachycardia (72%) since childhood and in all stages of life. Concealed accessory pathway reentrant tachycardias were identified in 22% of tachycardias with a stable incidence during the course of life. Various mechanisms of tachycardia were identified in the remaining patients. In conclusion, the mechanism of paroxysmal junctional tachycardia is not correlated with the age of the patient. PMID- 12555491 TI - [Efficacy of and tolerance to prolonged release prazosin in patients with hypertension and non-insulin dependent diabetes]. AB - The treatment of hypertension represents one of the major elements of the cardiovascular prognosis in type II diabetes. Among antihypertensive drugs, alpha blockers may be interesting because of the absence of unfavourable effects on plasma glucose and lipid levels. OBJECTIVE: The aim of this study was to evaluate the effectiveness and the safety of prazosin osmotic tablet treatment in non insulin-dependent diabetic patients with mild to moderate arterial hypertension. METHODS: After an initial 4-week-single-blind placebo period, 81 hypertensive subjects (162 +/- 11/96 +/- 5 mmHg) with type II diabetes were included in the study to receive prazosin osmotic tablet (o.t) open-label therapy at the dose of 2.5 mg/day for 12 weeks. After 4 weeks of treatment the dosage of prazosin o.t was increased to 5 mg/day if the diastolic blood pressure remained > or = 90 mmHg. RESULTS: Both supine and standing systolic and diastolic blood pressures were significantly decreased (P < 0.001) with prazosin therapy from 162 +/- 10/96 +/- 5 mmHg in supine and 160 +/- 12/95 +/- 6 mmHg in the upright position, to 149 +/- 15/86 +/- 9 mmHg and 148 +/- 16/86 +/- 9 mmHg respectively at the end of the 12-week-treatment period. There were no significant changes in the glycemic parameters (glycemia, haemoglobin A1c) during the prazosin therapy compared with baseline values. A significant decrease of triglycerides (P = 0.005), total cholesterol (P < 0.001) and LDL cholesterol (P = 0.03) levels was observed during prazosin therapy compared with the baseline measurements, whereas HDL cholesterol remained stable. Only 6% of the patients reported adverse events in relation with the study drug during the active treatment period. CONCLUSION: This study showed a significant decrease of the blood pressure in hypertensive subjects with type II diabetes after prazosin o.t treatment, without any change of glycemic parameters. Moreover, there was a favourable evolution of the lipidic parameters during the study characterised by a significant decrease of triglycerides and total and LDL cholesterol. PMID- 12555492 TI - [Short-term course under beta blockers of clinical and echocardiographic parameters in mitral stenosis in sinus rhythm]. AB - The purpose of this study is to evaluate the short-term benefit of a beta-blocker (atenolol) on clinical and echocardiographic parameters of patients presenting isolated or predominant mitral stenosis in sinus rhythm. It is a prospective study performed on 26 patients who have had a clinical and echocardiographic assessment before and 15 days after treatment by atenolol. After 15 days of beta blocker treatment, there is a significant improvement of dyspnea (57.6% in class III or IV before beta-blockade versus 15.3% with atenolol; P = 0.001) and a significant decrease of the heart rate (83.3 +/- 15.2 versus 68.9 +/- 13.9; P = 0.001) and the diastolic blood pressure (8 mmHg +/- 1.3 versus 7.2 mmHg +/- 0.9; P = 0.01). The Doppler echocardiography shows a significant increase of the stroke volume calculated by the Doppler method (28.7 +/- 6.2 versus 38.6 +/- 9.7 mL; P = 0.04). There is an insignificant trend to an improvement of the left ventricular systolic function, an increase of cardiac output and the decrease of the mean transmitral gradient. The factors associated with the failure of beta blocker treatment are: the right heart failure (P = 0.04) and the low diastolic blood pressure (P = 0.01). The beta-blockers could be a logical and effective treatment of patients with mitral stenosis waiting for balloon commissurotomy or surgery. PMID- 12555493 TI - [Isolated primary streptococcal tricuspid endocarditis. A case report]. AB - The authors report a new case of primitive isolated endocarditis of the tricuspid valve. The microorganism responsible, Streptococcus sanguis, is unusual in its localization and in this environment. The uncomplicated evolution poses the problem of deciding what action should be taken vis-a-vis the persistence of an important isolated vegetation after effective antibiotic treatment. Therapeutic abstention was chosen rather than surgical intervention in light of the literature. A spontaneous pulmonary embolic migration of the vegetation was effectuated 3 weeks after termination of the antibiotic treatment without pulmonary or hemodynamic complications. PMID- 12555495 TI - [When should mitral valve prolapse be considered a real disease?]. AB - Mitral valve prolapse should be considered as a disease when superior displacement of the mitral leaflets during systole is more than 2 mm with a maximal leaflet thickness of at least 5 mm. Using these criteria, the prevalence of mitral valve prolapse is 1.3% in the general population, nearly the same in men and women. Serious complications may occur during follow-up with a 1 to 3% patient-year risk. The most important complication is mitral regurgitation, mainly due to rupture of the chordae tendineae, which must be quickly corrected by surgical repair. Second is infective endocarditis, a complication which may occur particularly in men older than 45 years of age with systolic murmur. Arrhythmias are not infrequent but ischemic neurologic events are unusual, especially in young patients. Cases of sudden death have occasionally been reported. PMID- 12555494 TI - [Dilated cardiomyopathy secondary to acromegaly]. AB - In acromegaly, dilated cardiomyopathy is rare. Indeed, arterial hypertension, coronary artery disease and hypertrophic cardiomyopathy are more common. The authors report a case of a 42-years-old man with dyspnea (class III NYHA) and palpitations. The cardiac echography showed a dilated cardiomyopathy. The investigations revealed an acromegaly as an etiology. Diagnosis was based on patient morphotype, a high level of growth hormone (GH) and a pituitary adenoma detected by magnetic resonance imagery. This case is characterized by the absence of arterial hypertension and hypertrophic cardiomyopathy. The coronary angiography was normal. Consequently, in this particular case, dilated cardiomyopathy seems specific to acromegaly. PMID- 12555496 TI - [Radioclinical profile of cardiopericardial hydatid: report of 17 cases]. AB - Hydatid cysts concerning the heart are rare, accounting for 0.5 to 2% of all hydatic sites. The risk of serious complications in this location makes rapid diagnosis and surgical treatment essential. The aim of our study is to clarify the role of imaging in the diagnosis of the disease and to propose an adequate strategy. We report on 17 patients who underwent surgery for cardiopericardial hydatid cyst in the cardiovascular and thoracic unit of Sahloul hospital in Sousse from January 1988 to December 1998. Clinical investigation included in all cases chest X ray, ECG and ultrasonography (US). A computed tomography (CT) scan was performed in 14 cases, magnetic resonance imaging in three cases, transesophageal US in five cases, and coronary angiography in fsix cases. Examination for other hydatic sites was realized in all cases, and brain CT was performed in four cases. The hydatid cyst was variably localized in the left ventricle wall (five cases), the right ventricle (five cases), the pericardium (five cases), the interventricular septum (four cases), the right auricle (one case) and the left auricle (one case). The existence of other cardiac hydatid sites was found in 12 cases. All patients underwent surgery. Outcome was favorable in 14 cases, with a mean of 3 years and 5 months survey. Three patients died. The combination US-CT scan allowed a precise topographical inventory, reducing the need for MRI to the complicated cases and to the rare cases of inconclusive results by US-CT scan. PMID- 12555497 TI - [Validation of the therapeutic role of amlodipine in 31,946 French hypertensive patients]. AB - Amlodipine, a dihydropyridine calcium channel blocker (CCB), with a long duration of action, has been the subject of numerous controlled studies which showed its effectiveness and good tolerance in arterial hypertension in once-daily doses. We report the results of a large, multicentric, French, prospective phase IV study which evaluated the effectiveness and tolerance of amlodipine administered at a rate of 5 to 10 mg in only one daily dose. We also assess the evolution of the quality of life after 12 weeks of treatment among 31,946 hypertensive patients followed up to the ambulatory stage by general practitioners. The response rate- defined as the patients having had a reduction of 10 mmHg or more diastolic blood pressure--was 88%. The blood pressure standardization--defined by a diastolic blood pressure lower than 90 mmHg--was achieved for 70% of the patients. Amlodipine was administered in stand-alone therapy in 78% of the cases. The occurrence of an undesirable event was noted in the course of treatment in 12% of the patients and justified interruption of the treatment for 3.7% of the total population. The index average of quality of life was improved by the end of the 12-week treatment. This study carried out on a significant number of hypertensive patients (n = 31,946) under real prescription conditions confirms the efficacy and good tolerance of amlodipine, as has already been demonstrated in the preliminary developmental studies. PMID- 12555498 TI - [Buerger's disease or thromboangiitis obliterans]. AB - The Buerger's disease or thromboangiitis obliterants (TAO) is a non atheromatous inflammatory disease which alters medium and small-sized arteries and veins. It can be found world-wide, but it is more frequent in Eastern Europe, Middle East, Asia and Southeast Asia. Young men and smokers are the most affected. The incidence of this disease is increasing among women. The cause of this disease is unknown yet. The most striking fact is the relationship between TAO and tobacco. The diagnostic is most often late in front of a digital leg ischemia. Complementary exams help to its diagnostic and management but none of them are specific out of the pathology. The affection is evolving towards distal gangrene with amputation in 5 to 10% of cases. Prostacyclin demonstrated its efficiency. Revascularization surgery is difficult but sometimes possible. Sympathectomy, medular stimulation must be suggested. The patient survival is not at stake and the prognosis is above all functional. The most important element in the treatment is stop smoking definitively. PMID- 12555499 TI - [Current role of angioplasty in lower limb arteriopathy obliterans]. AB - For intermittent claudication caused by aorto iliac artery occlusive disease, percutaneous transluminal angioplasty is a safe and successful treatment. The introduction of vascular stents has been helpful in case of insufficient luminal gain (recoil or local dissection) or complications (extensive dissection). The results of angioplasty of femoropopliteal lesions must be compared with those obtained with medical or surgical treatment. In case of significant stenosis or short occlusions, conventional angioplasty alone is employed most of the time. Interventional radiology has an increasing role in the treatment of patients with critical ischemia. The majority of the culprit lesions are infra-inguinal. The goal of endovascular treatment is limb salvage (obtained in more than 80%) of the cases, not long-term patency. The aim of the procedure is to obtain a "straight line flow to the foot" by treating all the significant stenoses/short occlusions. PMID- 12555500 TI - [Which indications and techniques for lower limb arteriopathy obliterans and proximal aorto-iliac lesions?]. AB - Atherosclerotic aorto iliac occlusive disease are not the most frequent lesions in the lower limbs. Isolated aortic iliac lesions are often associated with intermittent claudication; critical ischemia with proximal aorto iliac lesions can be due to acute occlusion or associated femoropopliteal occlusive lesions. Preoperative explorations need Duplex Scan, Arteriography and CT Scan to define extensive lesions and wall' calcifications. Endovascular procedures are, in this lesion, safe, effective and permit to delay conventional surgery. The best strategy, today, needs to known endovascular procedures and surgical procedures to obtain the most efficient anatomic result. PMID- 12555501 TI - [Doppler echocardiography and lower limb arteriopathy obliterans]. AB - Duplex Doppler ultrasonography became the first line investigation in patients with lower limb arterial disease. It use the ultrasonic technology coupled to computer. Constant progress of duplex Doppler ultrasonography, as color flow imaging or power Doppler and more simply continuous refinements in grey scale image resolution, associated with the expansion of the interventional endovascular processing explains its significant development these last years. This simple non invasive technique is now an essential complement of physical examination. It is thus an usual tool use in common practice by vascular physicians and surgeons from tracking till processing while passing by the diagnosis. Duplex Doppler ultrasonography gives accurate informations about both structural and dynamic anomalies in the peripheral lower limb. It thus allows the evaluation, the quantification and the follow-up of the arterial diseases by carrying out a precise vascular mapping that can guides the radiological or surgical processing if necessary. Duplex Doppler ultrasonography remains however "operator depending" and moreover cannot replace the appropriate clinical decision making and medical therapeutic strategy. PMID- 12555502 TI - [Epidemiology of and risk factors for lower limb arteriopathy obliterans]. AB - Peripheral arterial disease (PAD) is most frequently caused by atherosclerosis. The prevalence of intermittent claudication is about 3 to 6% of the men over 60 years. But recent epidemiological studies demonstrate that prevalence of asymptomatic forms is two to three fold higher--about 3.4 to 12.1%. The most relevant diagnostic criteria for asymptomatic PAD is probably the measure of the ankle/brachial index. Most important risk factors for PAD are smoking and diabetes mellitus. Improving symptoms or stabilization are a common fate (50%) in PAD. About 25% will experience revascularization procedures (angioplasty or surgery), 4% will have severe amputation and worsening symptoms occur in 15% of patients. Critical limb ischemia is mostly rare (1%). It is not surprising that fate of claudicant and mortality is determined by coronary events and cardiovascular diseases. PMID- 12555503 TI - [Aortic dissection among blacks. Report of 6 Congolese cases]. AB - INTRODUCTION: The authors report on six cases of aortic dissection (AD). The work intended to bring out AD characteristics and attempt to define a profile of African patients who manifested this disease. The diagnosis of AD was determined on the basis of: clinical and radiological criteria (six cases); echocardiographic data (five cases); aortographic and surgical informations (two cases). PATIENTS AND METHODS: There were three males and three females with an average age of 40.2 years, with a variation of +/- 10.6 years (from 17 to 49). The frequency of AD was estimated at 1.8/10,000 admissions considering all causes and 5.4/10,000 for cardiovascular disease. Strain was the only triggering factor- noted in two cases (Observations 2 and 3). All patients suffered from severe arterial hypertension (AHT) of which there was one case of renovascular AHT based on the Takayasu arteritis. All symptoms and physical findings were the same described in literature. Operations were successful on two patients. The four unoperated patients died (tamponade: two cases, undetermined cause: two cases). CONCLUSION: The authors conclude as to the similarity of the AD charts described in the literature. However they stress the almost exclusive role of AHT in African studies series as well as that, in rare cases, of Takayasu's disease. They recommend preventing AD through the sustained and effective treatment of AHT. PMID- 12555504 TI - [Aneurysm of the internal saphenous vein: report of a case]. AB - Venous aneurysms of the lower extremities are rare. They have been reported in both the deep and superficial vein system. Thromboembolism is more common in aneurysms involving the popliteal vein than those involving superficial veins. We report a case of primary aneurysm of the greater saphenous vein presenting as an inguinal hernia. Sonography has confirmed the venous dilatation. we review also the clinical and therapeutic findings of venous aneurysms of the lower extremities. PMID- 12555505 TI - [Ruptures of the posterior wall of the left ventricle after mitral valve replacement: surgical repair and prevention]. AB - We report two cases of type III rupture of the left ventricle posterior wall, situated midway between the mitral annulus and the posterior papillary muscle and occurring in immediate continuations of a mitral valvular replacement. The first case has occurred at the moment of the canulas removal in a 28 years old patient operated for mitral stenosis. The tear was repaired with horizontal mattress sutures through felt strips on the external surface of the heart. The immediate post operative course was marked by bleeding recurrence and cardiac failure which has led to death. In the second case, a 68 years old man was operated for degenerative mitral insufficiency. The rupture has occurred just before closing the chest. The repair was realized successfully by using an intra-ventricular patch. The chordae tendinae removal constitutes a factor predisposing to the left ventricle posterior wall rupture. The repair has to use an intra-ventricular patch. The prevention is based on the preservation of the continuity between the mitral annulus and papillary muscles by preserving the sub-valvular apparatus or by using synthetic chordae. PMID- 12555506 TI - [Endocavitary implantation of a permanent pacemaker in the contralateral side of right pneumonectomy: a case report]. AB - The definitive endocardial stimulation is easy to install, allows a stable position of the leads, and a satisfactory stimulation thresholds for a long period. The epicardial approach is reserved for some rare indications including infectious contexts. The endocardial approach has been considered for a 67 years man with a complete AV block and an atrial fibrillation. This patient had undergone a right pneumonectomy 15 years before. A VVIR pacemaker has been implanted successfully by an internal jugular vein approach, and connected to a passively fixed unipolar lead. Because of the right ventricle deformation which made it unrecognizable, even by angiography means, we had to face major difficulties to position the lead. The epicardial approach should be considered even if a direct unique lung controlateral approach is easier than homolateral, because it allows us a quick ventricular access under the view control. PMID- 12555507 TI - [Stress cardiomyopathy: report of 2 cases]. AB - The authors reported two cases of acute catecholamines cardiomyopathy expressed clinically by chest pain and dyspnea, without any previous cardiac history. The diagnosis of human stress cardiomyopathy is established on: typically rapid onset aftermath of intense emotional stress, left ventricular apical akinesis and hyperkinetic motion of basal walls imaged by two-dimensional echocardiography. Rapid reversal clinical course and normal coronary arteriography. Similar finding have been observed in conjunction with pheochromocytoma who have been excluded by normal levels of urinary catecholamines metabolites. PMID- 12555508 TI - [Is thiamine supplementation necessary in patient with cardiac insufficiency?]. AB - Interest has recently risen regarding thiamine deficiency in patients with cardiac deficiency who are receiving long-term diuretic therapy. Thiamine deficiency can lead biventricular myocardial failure (cardiac beriberi), and treatment consists of thiamine administration. Studies have shown that long-term furosemide use may be associated with thiamine deficiency through urinary loss, contributing to cardiac insufficiency in patients with congestive heart failure. Thiamine supplementation could improved left ventricular function. However, the results of those studies are controversial, and none study have till proved the clinical impact of a systematic administration of thiamine in a cohort of patients with cardiac insufficiency. To date, and waiting for available literature, thiamine administration should be consider in patients at risk for thiamine deficiency (elderly, malnourished, alcoholic), and in patients receiving very large doses of diuretics. PMID- 12555509 TI - [Comparison of the antihypertensive efficacy of and tolerance to 2 imidazoline receptor agonists: moxonidine and rilmenidine in monotherapy]. AB - Two hundreds mild to moderate hypertensive patients (mean age 54 +/- 10, 115 males, 85 females) were randomised in a multicentre, double blind, two parallel groups trial in order to compare the efficacy of moxonidine (0.2 mg od) and rilmenidine (1 mg od), two selective I1 receptor agonists. The dosage had to be shifted after a four-week treatment period (moxonidine 0.4 mg od or rilmenidine 2 mg bid) if DBP > 90 mmHg. More than half of the patients had to be shifted to a double dose. The blood pressure was normalised (DBP < or = 90 mmHg) for 47% of the moxonidine patient's and 50% of the rilmenidine patients. The DBP decrease reached 7.3 mmHg in the moxonidine group and 8.0 mmHg in the rilmenidine group (P = 0.28). The SBP decrease was the same in the two groups (7.6 mmHg). Both drugs were well tolerated. The great affinity of moxonidine and rilmenidine for the I1 receptors and the weak affinity for the alpha 2 receptors were probably responsible of the small number of adverse events which are generally linked to the centrally acting antihypertensive drugs (asthenia, somnolence, oedema). Because moxonidine and rilmenidine are often to be prescribed at double dose and are often associated with other antihypertensives, moxonidine whose dosage could be shifted without having to increase the number of daily intakes could enhance the patient's comfort and make the compliance easier. PMID- 12555510 TI - [Effect of naftidrofuryl on physiological walking distance in patients with intermittent claudication]. AB - PURPOSE: To evaluate the physiological walking distance measured with the Peripheral Arterial Disease Holter Control device (PADHOC) after 12 months of treatment with naftidrofuryl in a double blind placebo controlled, parallel group study, in patients presenting with intermittent claudication. MATERIAL AND METHODS: The outpatients selected were of both sexes, aged 40 to 80, with a chronic, stable intermittent claudication and an ankle brachial index between 0.60 and 0.90. They received naftidrofuryl 200 mg tid or placebo for 12 months. Outcome measures included physiological painfree and maximal walking distances using the PADHOC device. The principle of this device is the measurement of the intermalleolar distances using ultrasound telemetry. The PADHOC measures the walking distance and the speed profile in an ambulatory subject. RESULTS: 182 patients were randomised and 168 entered the intention to treat analysis. The two groups were well matched for demographic variables, risk factors and history of vascular disease. After 12-month treatment, patients who received naftidrofuryl had a 107% improvement of geometric physiological pain-free walking distance versus 12% in the placebo group (P < 0.001) and 74% improvement of geometric maximal physiological walking distance versus 1% in the placebo group (P < 0.001). CONCLUSION: This study demonstrates the efficacy of naftidrofuryl versus placebo in patients with intermittent claudication using a new device measuring the walking distances of the patients in a more physiological way than the treadmill test. PMID- 12555511 TI - [Frequency of silent and painful ischemia in patients with treated stable coronary insufficiency]. AB - The present study was aimed at determining the frequency and circadian variations in symptomatic or silent myocardial ischemia in ambulatory patients with stable coronary disease. A comparative analysis was then made of the recordings on symptomatic and asymptomatic patients according to their medical history. Three hundred and twenty-one cardiologists recruited a total of 1,088 patients who were monitored for 4 days with a new type of electrocardiographic recorder. The patients were able to voluntarily start up the recorder in the case of cardiac discomfort or pain. The results showed that over a total recording period of 95,725 hours, the following data, which were validated by an experienced cardiologist, were obtained: 3,258 ischemic episodes, 2,963 (or 91%) of which were cases of silent ischemia, and 295 (or 9%) which were symptomatic. All the ischemic episodes involved a limited number of subjects, i.e., 271 patients. Of these, 148 (54.6%) were completely asymptomatic; only 63% of these patients with silent ischemia would have been detected if the recording had just lasted 24 hours. Moreover, the medical history showed a correlation between certain factors (such as poorly managed arterial hypertension, cardiac insufficiency, renal failure, arteritis of the lower limbs, and a waist-hip relation of over one in men) and an increase in the number of cases of silent ischemia. However, no single factor was found to be linked more to silent ischemia than to symptomatic ischemia. This investigation therefore shows the significant numeric incidence of silent ischemia. It raises the question of the need to prescribe treatment in at risk subjects which includes recordings of long duration, so that silent ischemia, which may increase the risk of mortality, can be more readily detected. PMID- 12555512 TI - [Endomyocardial fibrosis: report of 22 Congolese cases]. AB - A study has been made of 22 cases of endomyocardial fibrosis (EMF) observed over a 12-year period. The epidemiological characteristics have been described, and the importance of the echocardiographic findings in the diagnosis of the disease has been emphasized. EMF constitutes 0.3% of the total admissions in the department, 0.9% of the cases of cardiac failure, and 3% of the cases involving subjects under 40 years old. The patient population consisted of 13 men and 9 women with an average age of 35.6 +/- 16.4 years (age range: 8.5-77 years). The diagnosis of EMF was based on clinical, radiological, electrocardiographic, and echocardiographic findings (22 cases), and surgical examination (1 case). In 8 cases, parasitosis (filariasis: the patients came from a known endemic area) with hypereosinophilia was observed. Three of these patients had associated high blood pressure. In 13 subjects, a severe right adiastolic syndrome was noted. Two patients presented with tachycardia, 2 others had signs of overall cardiac failure, and 5 subjects suffered from palpitations connected with arrhythmia. Thoracic X-ray showed cardiomegaly in all cases, lung involvement in 15 cases, normal lungs in 5 cases, and bilateral hilar stasis in 2 cases. Electrocardiography mainly showed endomyocardial fibrosis (15 cases), supraventricular arrhythmia, notably auricular fibrillation (13 cases), and conductive disorders (12 cases), which were frequently associated. Echocardiography showed the presence of EMF in 21 cases (95.5%). Doppler (n = 9 cases) detected tricuspid failure in 9 subjects, and mitral failure in 1 subjects. EMF was exclusively located on the right side in 19 cases, bilateral in 2 cases, and on the left in 1 case, which required surgery. Four patients died (i.e., 1 case of sudden death, 1 case of pulmonary embolism, 1 case of neurological coma, and 1 case of cardiac arrhythmia). The authors, like many others, note the clinical polymorphism of EMF, the predisposition to the disease caused by the presence of helminthiasis, which should be eradicated, the diagnostic value of echocardiography-Doppler, and the efficiency of surgery in the treatment of this condition. PMID- 12555513 TI - [Treatment of aorto-coronary saphenous vein bypass lesions: pros and cons of angioplasty]. AB - Heart patients who have undergone a coronary bypass may present with renewed myocardial ischemia, often connected with bypass dysfunction. The saphenous bypass is the most frequently implicated, and palliative revascularization may be envisaged, either by further bypass surgery, or by angioplasty. The latter approach has been developed since the beginning of the 1980s, and since that time there has been considerable technical and pharmacological progress in performing this type of graft. However the indications for angioplasty and its comparison with reoperation remain controversial. In the present study, it therefore seemed pertinent to include the respective opinions of two experts in the field of angioplasty and coronary bypass surgery. PMID- 12555514 TI - [Tachycardia-induced cardiomyopathy, unusual and reversible cause of left ventricular dysfunction: report of 9 cases]. AB - In this study, 9 cases of tachycardia-induced cardiomyopathy have been reported, with a 7-year follow-up period. The patient population consisted of 5 males and 4 females with a mean age of 41 +/- 18 years (range: 10-70 years). It was difficult to determine the onset of the arrhythmia, due to its frequently asymptomatic course until such time as cardiac insufficiency became apparent (this was the case in 7 patients, while 2 subjects had palpitations). Four cases of AV intranodal reentry tachycardia (3 with an accessory pathway), 2 cases of atrial fibrillation, 1 case of auricular flutter, 1 case of atrial tachycardia and 1 case of ventricular tachycardia were observed. Disappearance of the symptomatology following restoration of sinus rhythm was accompanied by echocardiographically-determined normalization of systolic function on average 5 +/- 4 months post-arrhythmia (range: 1-12 months). Left-ventricular end-diastolic volumes decreased from 60 +/- 5 mm to 51 +/- 4 mm (P < 0.01), and end-systolic volumes from 50 +/- 7 mm to 35 +/- 4 mm (P < 0.001), while the left ventricular shortening fraction increased from 17 +/- 5% to 33 +/- 4% (P < 0.0001). In the differential diagnosis for primary dilated cardiomyopathy, tachycardia-induced cardiomyopathy appears very similar to the former, and only an adequate period of follow-up after the restoration of sinus rhythm can confirm the diagnosis. PMID- 12555515 TI - [Intraventricular thrombosis complicating peri-partum idiopathic myocardiopathy]. AB - Ventricular thrombosis can complicate the development and worsen the prognosis in any case of hypokinetic dilated cardiomyopathy. In the present article, a study has been made of 6 reports of ventricular thrombosis selected out of 58 medical files on women with peri-partum idiopathic cardiomyopathy. Patient age ranged from 22 to 55 years. The clinical picture showed hypokinetic dilated cardiomyopathy, complicated by cardiac failure; with its onset during the last trimester of gestation or in the 6 months post-partum. In all patients, overall cardiac failure was observed, and in all cases the diagnosis of intracardiac thrombosis was made by echocardiography. In all 6 patients, a left ventricular apical thrombosis was detected. In 2 subjects, 2 and 3 left ventricular thrombi were respectively found. In 1 case, a left ventricular thrombosis was present. In another case, a right thrombosis associated with a left ventricular thrombosis was detected. Treatment was initiated with a combination of anticoagulants (heparin and K antivitamins), diuretic and vasodilatory treatment. The clinical outcome was favorable, with the disappearance of thrombi and signs of cardiac failure (between the 15th and 54th day). No embolic complication was observed. These findings clearly show the importance of prescribing an anticoagulant treatment as a preventive measure during PPICM. Even if severe embolic complications are a potential risk, anitcoagulant treatment can ensure a favorable outcome. PMID- 12555516 TI - [Comparison of tissue harmonic imaging and standard imaging in echocardiography based on 4 different emission frequencies]. AB - Harmonic imaging is an innovative technique which is now available in most of the echographic equipment currently on the market. The use of echographic contrast agents led to the introduction of this particular technology, which has largely superseded the former method. In everyday medical practice, it seems to provide a better definition of endocardial contours. In this article, the authors have described a prospective study including 32 patients with an average age of 64 years (+/- 5 years). The aim was to compare imaging quality obtained by 2 acquisition frequencies for standard imaging with that procured by harmonic imaging (emission and reception frequency, 2.5 and 3.3 MHz respectively for the former technique; emission and reception frequency, 1.7 and 2.2 MHz respectively for the latter). The overall scores given by 2 different examiners for emission and reception frequency were as follows: 3.28 and 3.15 for 2.5 MHz, 2.84 and 2.56 for 3.3 MHz regarding the standard technique; 2.56 and 3.031 for 1.7 MHz and 2.56 and 2.063 for 2.2 MHz for the harmonic imaging technique. The superior performance of tissue harmonic imaging has been confirmed in the endocardial study of the left ventricular segments and in the analysis of the mitral valve. This study shows that harmonic imaging, a technique which was easy to use in the present study, provides a means of obtaining an average level of ultrasound imaging that facilitates the interpretation of the clinical picture. PMID- 12555517 TI - [Tolerance and efficacy of carvedilol prescription in patients with mild to moderated chronic heart failure]. AB - The current treatment for heart failure, i.e., carvedilol administration, has brought about a significant reduction in cardiovascular morbidity and mortality. The European Cardiology Association task-force has recently recalled that the treatment for heart failure should be considered differently according to the regions within Europe. However, most studies have involved North American or Australasian subjects. The results of these studies have shown that carvedilol improves the symptomatic status of patients with mild to moderate chronic heart failure, but their extrapolation to the situation in Europe is questionable. In the present study, the authors have reported on the use of this beta-blocker over a 6-month period in 48 European subjects (an older patient population, and more frequently given digitalis treatment) with mild to moderate stage II and III chronic heart failure (mean NYHA functional stage = 2.75 +/- 0.2; mean left ventricular ejection fraction = 32.4 +/- 3.4). Six patients (12.5%) were obliged to withdraw from the study due to adverse reactions to carvedilol, i.e., 2 subjects during the test-dose (4.2%) and 4 others who were subsequently unable to withstand the progressive stepping up of drug dosage (8.3%). In the remaining 42 patients, the mean NYHA scores dropped significantly from 2.75 +/- 0.2 to 1.8 +/- 0.1 (p < 0.05) at the end of the study period, indicating an increase in symptomatic improvement with a favorable hemodynamic tolerance profile, even in cases of combined therapy with digitalis. This study confirms the positive action of carvedilol on functional symptomatology in southern European patients presenting with mild to moderate chronic heart failure. However, it should be emphasized that this beta-blocker should only be administered under strict and specialized medical surveillance as its effect of significantly slowing down the heart rate could also result in cardiac insufficiency. PMID- 12555518 TI - [A study on taxonomy of rhizobia isolated from Kummerwia sp. and Indigofera sp]. AB - The rhizobial strains isolated from Kummerowia and Indigofera and the known reference strains were classified by performing numerical taxonomy. New isolated strains were divided into two new clusters at 83% similarity level. Based on the numerical taxonomy, additional isolates in each cluster were studied by using SDS PAGE of whole-cell protein. Twenty-four strains isolated from Kummerowia fell into cluster 1. Twenty strains isolated from Indigofera fell into cluster 2. The results of G + C mol% and DNA homology analysis showed that the DNA homologies between the central strain SH713 and SHL042 and the 13 type strains were less than 61%. Thus, the rhizobial strains from Kummerowia and Indigofera were two new individual species of Rhizobium. PMID- 12555519 TI - [Characterization and phylogenetic analysis of the new species of the Leucobacter tropicalis sp. nov]. AB - A new aerobic, gram-positive, non-sporulating, rod-shaped organism is described. Strain F-8 was isolated from tropical soil, and has the following characteritics: the menaquinone contains a side chain with 11 isoprenyl units(MK-11); 2,4 diaminobutyric acid, glutamic acid, alanine, glycine and a small amount of gamma aminobutyric acid present in the cell wall at a molar ratio of 0.77:1.0:1.66:0.95. The major cellular sugar were rhamnose, galactose and glucose. The G + C content of DNA is 68 mol%. The 16S rRNA gene was amplified, cloned and sequenced, a phylogenetic tree was constructed on the 16S rRNA gene sequences. The tree clearly indicated that strain F-8 forming the same lineage with leucobacter komagatae, the level of binary sequence similarity between strain F-8 and L. komagatae is 96%. Based on the characteristics, allow the placement of strain F-8 into the genus Leucobacter, but F-8 showed some differences in biochemical and physical characters from the reported species L. komagae. A new species name, Leucobacter tropicalis sp. nov. was proposed for this isolate. The type strain is designated F-8. PMID- 12555520 TI - [Immunocytochemical identification of tropomyosin in nucleoli and nucleolar matrix of Physarum polycephalum]. AB - Nucleoli were isolated from physarum polycephalum, and nucleolar matrix was prepared by digesting the nucleoli respectively with DNase 1, 0.25 mol/L (NH4)2SO4 and 2 mol/L NaCl to remove DNA and most proteins. SDS-PAGE analysis indicated that there were about 20 polypeptides in nucleolar matrix component, including the 37 kD polypeptide which was similar to tropomyosin in molecular weight. The result of indirect immunofluorescence treated with anti-tropomyosin antibody and sheep anti-rabbit IgG antibody labelled with FITC showed that bright fluorescence was observed in the nucleoli and nucleolar matrix, but no bright fluorescence in the controls. Indirect Immunoblotting detection further verified that tropomyosin existed in nucleolar matrix. Protein A-colloidal gold immunoelectron microscopic study showed that there were many gold particles in the specimens labelled with tropomyosin antibody, and there were few gold particles found in the controls. Tropomyosin distributed dispersedly in nucleoli. PMID- 12555521 TI - [Construction of a GFP broad-host-range promoter-probe vector and its application]. AB - The 0.9 kb DNA fragment was obtained by PCR and using a pair of special designed primers from the green fluorescent protein gene(gfp). It Was then cloned into an expression vector pET-11C and named as pHN115. A promoferless gfp vector with SD fragment was obtained from pHN115 and subcloned into SK(+), pIJ 2925 and finally into pTR102. A broad-hast-range and visible promoter-probe vector, pHN127 was constructed and used successfully for the identification of constitutive and inducible promoters from Sinorhizobium fredii HN01. PMID- 12555522 TI - [Cloning, sequencing and expression of Sinorhizobium meliloti strain 042B gene nodD]. AB - S. meliloti strain 042B is a rhizobium strain which can form nodules both on alfalfa and on soybean. In this study, nodD gene of 042B was cloned into pBBR1MCS 5. By functional analysis in the system of R. leguminosarum bv. viciae LPR5045 (pSym-, pMP154), it was found that the NodD has responses both to luteolin and to genistein. This result showed that the 042B nodD gene is probably a specific nodulation determinant which determined its capability of nodulation on both alfalfa and soybean. The nodD fragment was then cloned into the expressional vector pThioHis A, B and C, and three recombinant plasmids pXDA, pXDB and pXDC were constructed. The plasmid pXDC was identified to be in the same open reading frame with the trxA gene of pThioHis C throuth sequencing analysis. Inducing IPTG and analyzing with SDS-PAGE, it was found that the fusion protein expressed from E. coli Top10(pXDC) with the molecular weight of TrXA and NodD together. The Western bolt result demonstrated that the expression result is the target gene nodD product. PMID- 12555523 TI - [Location of the binary toxin gene of Bacillus sphaeriucs and some biological characteristics of its asprogenous mutants]. AB - Location of the binary toxin gene of Bs C3-41 and some biological characteristics of its asporogenous mutants which obtained by DES treatment were studied in this paper. Mutant C4, L5 with blocked at the stage II of sporulation course were not possessed of any crystalline inclusion, but the binary toxin proteins were detected in their cells by SDS-PAGE analysis. Another mutant G5 which blocked at the stage III of sporulation course formed parasporal crystal in the cells, and its toxicity to larvae of Culex quinquefasciatus was about 50 times higher than those of mutant C4 and L5. The result of curing plasmid and Southern blot analysis confirm the binary toxin gene of Bs C3-41, Bs 1691 and Bs-10 locate on the chromosome. PMID- 12555524 TI - [Expression of genes aroG and pheA in phenylalanine biosynthesis]. AB - aroG and pheA genes, encoding 3-Deoxy-D-arabinoheptulonate-7-phosphate synthase(DS) and Chorismate mutase (CM)-prephenate dehydratase(PD) in the pathway of phenylalanine biosynthesis respectively, were amplified by polymerase chain reaction(PCR). The genes were assembled on the multicopy vectors and expressed in both Escherichia coli and Brevibacterium. The products of two gene were detected by SDS-PAGE. The activities of relevant enzymes were measured in the crude extract of the host strain. When aroG-pheA genes were introduced into E. coli p2392, the activities of DS, CM and PD were increased by 4.3-fold, 4.4-fold and 2.2-fold respectively. Whereas in the case of Brevibacterium flavum 2732, the activities of DS, CM and PD were increased by 12.3-fold, 2.3-fold and 5.6-fold, respectively. As the results, the overproduction of phenylalanine was brought about by using the genetic engineering strain of B. flavum. PMID- 12555525 TI - [Gene regulation studies on purine biosynthetic in Salmonella typhimurium IX. Mutation analysis of PUR box 2]. AB - Two consensus bases C and G in 16 bp PUR box were directed mutated to G and A separately by PCR amplification. The binding function of PUR box carrying mutation with PUR protein were examined by gel retardation experiment. The results showed that the PUR box with above mutations could not bind with purR protein extracted from LT2. It proved that the two consensus bases C and G are necessary for PUR box binding with purR protein. PMID- 12555526 TI - [Purification and characterization of antifungal peptide LP-1]. AB - An antifungal peptide LP-1 from Bacillus subtilis TG26 strain was purified by acid precipitation, acetone precipitation and Hi-pore reversed phase column chromatography. The molecular weight of LP-1 is 1057.3 D as determined by MALDI TOF mass spectrometry, and its pI is 4.75 by PAG-IEF. It was also found to be thermostable. Its antifungal spectrum showed that LP-1 has strong inhibitory activity against many plant pathogenic fungi, such as Pythium aphanidermatum, Gibberella zeae, Alternaria longipe, Fusarium oxysporum f. lycopersici, etc.. The abnormal hyphal growth of Trichoderma viride caused by LP-1 such as swollen tips, twisted, short growth and cytoplasm condensation was also observed. Both ninhydrin reaction and peptide sequencing suggested that LP-1 is a cyclic peptide. PMID- 12555527 TI - [Studies on the metabolism pathway of aniline degradation by Comamonas acidovorans AN3]. AB - All of the enzymes relating to aniline degradation in C. acidovorans AN3 were inducible ones. When growing on aniline as sole carbon, nitrogen and energy sources, the cells of C. acidovorans AN3 contained aniline dioxygenase, catechol 2, 3-dioxygenase, 2-hydromuconic semialdehyde dehydrogenase, 4-oxalocrotonic acid decarboxylase and 4-hydroxy-2-oxovalerate acid aldolase. The Km and Vmax of aniline dioxygenase were 292 mumol/L and 3.57 mumol.mg-1. min-1, respectively. The Km and Vmax of catechol-2, 3-dioxygenase were 16.4 mumol/L and 15.2 mumol.mg 1.min-1, respectively. According to these results, the degradation metabolism pathway of aniline by C. acidovorans AN3 was hypothesised. PMID- 12555528 TI - [Breeding of yeast for beer manufacturing by inactivated protoplast fusion]. AB - QSB-XI6, one of fusants, was obtained by PEG-induced fusion between UV inactivated protoplasts of Saccharomyces cerevisiae LQ16 (Ile-, Datr) and heat inactivated protoplasts of S. cerevisiae QSB7(Ala-, H2S-). The cell volume, bicmass, genotype and DNA content of the fusant were measured in comparison with of its parents and the results showed that QSB-XI6 was a fusant. The fermentation ability and the flocculent capacity of this fusant were higher than those of its parents. The diacetyl content of the beer produced by the fusant was measured and some other compositions of the beer was also analysed by gas chromatography. The sensory evaluation of the beer was much better and the flavour was distinctive. The results of many production trials in succession showed that all good characteristics of QSB-XI6 were stable. PMID- 12555529 TI - [Antibiotic resistance and plasmid profiles of Vibrio isolates from cultured Sparus sarba]. AB - A total of 51 potential pathogenic vibrios were isolated from moribund silver seabream Sparus sarba, which were collected from local fish farms of Hong Kong. All the isolates were classified and identified as 7 species by the API 20 E system and the scheme of Alsina & Blanch. These species were Vibrio alginolyticus (24 strains), Vibrio vulnificus (12 strains), Vibrio parahaemolyticus(7 strains), Vibrio logei(4 strains), Vibrio pelagius II(2 strains), Vibrio fluvialis (1 strains) and Vibrio meditterranei (1 strains). Among these isolates, the three predominant species (V. alginolyticus, V. vulnificus and V. parahaemolyticus) were confirmed to be virulent to sea bream by experimental challenge. All isolates were also screened for plasmid DNA by agarose gel electrophoresis and tested for susceptibility to 16 antimicrobial agents by the agar dilution method. Of the 51 isolates examined, all strains were sensitive to ceftriaxone, streptomycin, nalidixic acid and rifampicin, and almost all were sensitive to ceftazidime, netilimicin, chloramphenicol and sulfamethoxazole except one or two strains. Most isolates were resistant to ampicillin (60. 8%), cefuroxime (66.7%), amikacin(55%), kanamycin(58.8%) and trimethoprinm (76.5%). Fifteen of the 51 isolates harboured 1-4 plasmids, with sizes ranging from 9 to 123 kb. Both the plasmids and the associated antimicrobial resistance (ampicillin, cefuroxime and trimethoprim) of 9 isolates could be transferred to recipient by single-step conjugation, however, the frequencies were very low, ranging from 10(-11) to 10( 9). The present results indicate that resistance to these antibiotics is chromosomal. PMID- 12555530 TI - [The effects of reactive oxidants on Bacillus thuringiensis parasporal crystals]. AB - The effects of hydrogen peroxides(H2O2) and hydroxyl radicals(.OH) on the purified parasporal crystals from Bacillus thuringiensis var. Kurstaki HD-1 were studied by SDS-PAGE and bioassay. The results showed that to some extent H2O2 and .OH produced a direct and deleterious effect. Hydroxyl radicals from the system Fe(II)-EDTA-H2O2 indicated more serious damage than hydrogen peroxides. There was a 44.5% loss in activity against 12-h-old larvae of Heliothis armigera when parasporal crystals were treated with 0.3% H2O2 only. However, 85.2% activity lost when parasporal crystals were treated with 0.3% H2O2 in the presence of iron(II) and EDTA. PMID- 12555531 TI - [Purification and properties of glutamate dehydrogenase from Pseudomonas pseudoalcaligenes]. AB - Glutamate dehydrogenase was purified from the crude extract of Pseudomonas pseudoal-caligenes. The enzyme had a molecular weight of 290,000 and was composed of six subunits with identical molecular weight of 47,000. The enzyme was highly specific for NADP(H) and the substrates. The biochemical properties such as kinetic parameters and heat stability were also examined. The purified GDH showed considerable loss of activity upon freezing. PMID- 12555532 TI - [Progress in research of Vitreoscilla hemoglobin and Vitreoscilla hemoglobin gene]. AB - Vitreoscilla is a Gram-negative obligate aerobic bacterium, whose most important property is the ability to produce the Vitreoscilla hemoglobin (VHb). VHb is encoded by a single gene called Vitreoscilla hemoglobin gene (vgb) containing the natural promoter, and the expression of vgb gene is regulated by dissolved oxygen at the level of transcription. The oxygen binding VHb participate in the metobolism process of cells and make them grow well in the hypoxic habitats. It also facilitates the bacteria growing and the potein producing obviously. The potential application prospect for the oxygen regulated promoter of vgb and the physiological function of VHb in the fermentation industry is reviewed. PMID- 12555533 TI - [The morphological structure of classical swine fever virus and some characteristics of its multiplication]. AB - Some characteristics of the multiplication of classical swine fever virus(CSFV) Thiverval strain were studied by means of the immunofluorescence technique. Under optimum culture conditions, the concentration of CSFV in the culture liquid multiplicated by MPK cells is ten times higher than by PK-15 cells. The half-life period of CSFV at 37 degrees C is 3 hour. The location of CSFV's replication in host cells is detected by the monoclonal antibodies of the structural protein E2 and the non-structural protein p120 of CSFV. Based on the above results, the ultrastructure of CSFV particles inside host cells was further studied using electron microscope and the changes of the ultrastructure of CSFV-infected cells are shown. The possible reasons for lower viral titre are also discussed in this paper. PMID- 12555534 TI - [Expression in Lactococcus lactis of catalytically active phenylalanine ammonia lyase from parsley]. AB - The phenylalanine ammonia-lyase (PAL) cDNA of parsley (Petroselinum crispum) was subcloned into constitutive expression vector pMG36e downstream of the P32 promoter. The resulting plasmid pMG36ePAL was introduced into Lactococcus lactis subsp. lactis MG1363 by electroporation. The recombinant strain showed its PAL activity conversing the L-phenylalanine in the culture medium into trans-cinnamic acid. A new secretory vector pXHS was constructed by recombination of pMG36e with a Lactococcal usp45 secretion leader coding sequence and a translational coupling sequence. Then the pXHSPAL was constructed and used for expression of PAL in L. lactis, the PAL activity was also detectable. The L. lactis dnaJ promoter sequence was cloned and used to construct a heat inducible vector pXHJ. PAL cDNA was cloned into pXHJ and the L. lactis IL1403 was transformed with the recombinant plasmid pXHJPAL. After a heat shock from 30 degrees C to 37 degrees C, the PAL activity of the pXHJPAL strain could increase approximately onefold. The prospect of using these engineering L. lactis strains for PKU therapy was also discussed. PMID- 12555535 TI - [Influence of EBV latent membrane protein 1 gene on the proliferation of epithelial cells]. AB - In order to study the influence of EBV latent membrane protein (LMP1) gene on the proliferation of epithelial cells and the role of LMP1 creating to epithelial cell carcinomas, EBV LMP1 gene eukaryotic expression plasmid was transfected into human embryo kidney epithelial cells. The expression of LMP1 protein in transfected cells was detected. Observing the formation of colonies in soft agar, the value of MTT absorbed and the expression of PCNA, the results showed that the transfected cells grew vigorously and formed more colonies in soft agar, the MTT uptake and PCNA expression were apparently increased. The conclusion was that LMP1 gene could alter the biological behaviours of epithelial cells, promote cell growth, proliferation and transformation and endowed the transfected cells with growth characteristics like tumor cells. PMID- 12555536 TI - [Expression detection and location analysis of BstNI isoschizomer restriction modification system gene]. AB - Some genetic markers of E. coli HB101 and JM110 were identified, two bacterial strains were used as recipients respectively to detect the expression of a restriction endonuclease(R) gene and a methylase(M) gene of BstNI isoschizomer restriction-modification system. DNA fragment containing the R-M genes was deleted unilaterally with exoIII and 23 deletion subclones were obtained. According to the Enzyme activity of each subclone, R and M gene were located respectively at the regions of 0.2-->1.4 kb and 1.5-->3.3 kb from cloning site PstI. Analysis showed that the R. M system belongs to type II, two genes are controlled by the different promoters; the recognition sequence of this system is the same as that of DNA-cytosine methyltransferase(Dcm), the latter's methylation function can resist the R enzyme. It was interesting that the recombinant plasmid with an R+ M- genotype appeared to be lethal to dcm+ hosts yet. This indicated that the M gene closely linking to R gene is of critical importance for the existence of the R-M system in process of evolution. PMID- 12555537 TI - [Molecular cloning of rat OB gene and its expression in Escherichia coli]. AB - In order to provide rat OB gene product for studying the relationship between obesity and noninfectious diseases, rat OB cDNA was amplified by RT-PCR technique. 460 bp fragment of OB cDNA was subcloned into EcoRI/BamHI site of plasmid pUC 19. Sequence analysis of OB cDNA revealed that the translation reading frame was identical with that reported in the literature. Thereby plasmid pBV220-rOB was constructed and the specific expression of OB gene in E. coli identified by SDS-PAGE electrophoresis was obtained. PMID- 12555538 TI - [Different protein forms of purple membrane produced by Halobacterium halobium under various nutritional condition]. AB - The extremely halophilic bacterium, purple membrane producing Halobacterium halobium strain R1 were cultured in four liquid media which consist of different constituents. After the purple membrane is harvested by ultracentrification and purified by sucrose density gradient ultracentrification, and run on SDS-PAGE, the result of Coomassie blue staining showed different bR protein forms: three bands with molecule weight ranging from 26.0 to 27.5 kD in peptone medium, while only one band with MW 26.0 kD in other three media-complex, synthetic and artificial seawater media, which corresponded to the mature form in peptone medium. The result of Western blotting not only confirmed the different protein forms of Coomassie blue staining in the four media, but also gave additional bands that Coomassie blue can not detect due to its lower sensitivity: 1) in the complex, synthetic and artificial seawater media, a faint 28.0 kD band existed, while in the peptone medium it did not exist; and 2) in the four media, it appeared that a faint 23.0-24.0 kD band exited. The different bR protein forms of purple membrane which result from the difference of nutritional constituents in these four media might be caused by different precursor processing enzymes or different activities of precursor processing enzyme(s) under these different nutritional conditions. PMID- 12555539 TI - [The outer membrane protein (OMP) patterns of Escherichia coli isolates of predominant O serogroups originated from chichens in different regions in China]. AB - The purpose of this study is to determine the outer membrane protein (OMP) patterns of avian Escherichia coli isolates with predominant serogroups originated from 18 provinces, autonomous regions and municipal cities in China. Total of 204 isolates belonging to O18, O78, O2, O88, O11 and O26 serogroups respectively, were tested. The outer membrane proteins of these isolates were isolated with the improved N-lauroylsarcosine method and analyzed by SDS-PAGE. 4 different OMP patterns were identified with these isolates of predominant serogroups. 3 different OMP patterns with 56 isolates of O18, 4 with 54 isolates of O78, 2 with 28 isolates O2, 1 with 26 isolates of O88, 3 with 22 isolates of O11 and 1 with 18 isolates of O26 respectively, were found. Isolates with OMP pattern 1 were discovered in 6 different serogroups, and OMP-3 pattern was also shared by O18, O78 and O11 serogroups isolates. These results indicated that the OMP patterns of avian pathogenic Escherichia coli isolates of O18, O78, O2, O11 serogroups which were isolated from different regions in China were heterogeneous, and all of O88 and O26 serogroups isolates just belonged to OMP pattern 1. Moreover, the OMP pattern 1 was presented in isolates of six different predominant O serogroups. PMID- 12555540 TI - [Prokaryotic expressed trichosanthin and other two proteins have anti-fungal activity in vitro]. AB - DNAs encoding Trichosanthin, tobacco class I Chitinase and tobacco class I beta 1, 3-Glucanase were expressed in E. coli, respectively. The expression products were assayed for their anti-fungal activity. All of three kinds of proteins show anti-fungal activity. When two of them combined, this activity was enhanced greatly. When three of them combined, the stronger anti-fungal activity was observed. PMID- 12555541 TI - [Analysis of rRNA gene restriction fragments length polymorphism of Leptospira in China]. AB - Sixty-four Leptospira international and domestic reference strains, which belonging to fifty-four serovars, and twenty-seven field strains were examined by using EcoR I restriction endonuclease analysis of genomic DNA and restriction fragments length polymorphism of rRNA gene, fifty-six Leptospiral ribotypes(RTs) were described. Most serovars gave specific patterns. Serovars in the same serogroup possess common core-segments, but we found RTs of reference strains from China and other countries are different. Most field strains have RTs with correspond of reference strains, only a few bands were shown different if RTs were different. A notable result was that the field strains of serovar pomona have the same Rt as the international reference strain but different from the domestic reference strain. PMID- 12555542 TI - [Optimization of fermentation conditions for the production of poly(3 hydroxybutyrate-co-3-hydroxyvalerate) with Alcaligenes eutrophus]. AB - A close relationship, between the initial addition time and concentration of propionate and HV fraction, was observed in shaking culture of Alcaligenes eutrophus for the production of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate). The optimal initial addition time of propionate was determined at the onset of PHBV formation period. Although relatively high HV unit could be obtained under high propionate concentration, the growth and product synthetic activities were inhibited obviously. Different ratios of glucose to propionate were fed to stimulate the formation of HV unit and the results were compared. The optimizing feeding strategy of propionate was proposed based on the consideration of several fermentation index. The final cell dry weight, PHB concentration, PHB content and HV fraction in PHBV reached 52.1 g/L, 40.8 g/L, 78.3% and 16.2 mol%, respectively, Yield coefficient of HV unit to propionate and PHBV productivity were obtained to be 0.5 g/g and 0.74 g/(L/h), respectively. PMID- 12555543 TI - [Light-dependent incorporation of selenite into selenocysteine by isolated chromatophore of Chromatium vinosum]. AB - Illiminated intact chromatophore of chromatium vinosum in the presence of O acetylserine(OAS) catalysed incorporation of SeO3(2-) into selenocysteine at rate of 359 nmol.mgBchl-1.h-1. Sonicated chromatophore catalysed SeO3(2-) incorporation at 1.1% of the rate of intact chromatophore. Addition of GSH and NADPH increased the rate to 88.3% of intact rate, but SeO3(2-) incorporation under these conditions was essentially light dependent. The purified GSH reductase from Chromatium vinosum in the presence of cysteine synthase OAs and NADPH catalysed incorporation of SeO3(2-) into selenocysteine. It is proposed that SeO3(2-) is reduced by light-coupled GSH reductase and that Se2- produced is incorporated into selenocysteine by cysteine synthase. PMID- 12555544 TI - [Studies on reduction of Au3+ by bacteria for preparing gold catalyst]. AB - The strain D01, screened from different source bacterial strains, has relatively strong ability of adsorbing Au3+. The strain D01 was identified as Bacillus megatherium D01 and still grew well in the medium containing 600 mg/L Au3+. The results of electrochemical reaction indicated that the strain D01 had a strong ability of reduction. It could reduce the Au3+/alpha-Fe2O3(precursor of the gold catalyst) to highly dispersive Au0/alpha-Fe2O3 catalyst which could catalyze the reaction of CO + O2-->CO2. PMID- 12555545 TI - [The isolation of microbe producing biosurfactants]. AB - Microorganisms capable of producing biosurfactants can be isolated by a series of steps including hydrocarbon enrichment culture, hemolytic activity assay on blood agar plates and oil displacement activity assay etc. Ten strains(including bacteria, yeasts and fungi) were isolated with higher surface activity from 1000 samples from soil, sludge, waste water etc. Two strains of bacteria produced rhamnolipids and trehaloselipids respectively. One strain of bacterium produced long chain unsaturated carboxylic acid and another produced azelaic acid. Two strains of yeasts produced lipopolysaccharides with excellent emulsification property. PMID- 12555546 TI - [Expression of a HCV multi-epitopes antigen gene and study on its immunogenicity]. AB - Due to the hypervariable character of hepatitis C virus (HCV), 5 conserved T and/or B cell epitopes from core, envelope, NS3 and NS5 protein of HCV were chosen to form a 270 bp multi-epitopes antigen gene. The gene was clone into a fusion vector pWR450-1 to express a beta-galactosidase-HCV hybrid protein GZ-PCX. The purified GZ-PCX protein was specifically recognized by human anti-HCV antibodies. These results show that the HCV hybrid multi-epitopes antigen has excellent immunogenicity, which might be able to be used as an effective diagnosis agent and to provide protectivity to any genotype of HCV which might partly solve the problems in the researches of HCV vaccines. PMID- 12555547 TI - [Studies on preparation of L-phenylalanine from phenylpyruvic acid by E. coli EP8 10]. AB - E. coli EP8-10 was selected from the soil. It was able to produce the transaminase with high activity when it was cultivated on the medium containing peptone and beef extract. Optimum conditions of enzyme reaction was: phenylpyruvic acid's concentration of 0.3-0.5 mol/L, L-Asptaric acid used as amino donor, pH 8.5 37 degrees C. When phenylpyruvic acid was 0.3 mol/L, 48 g/L L phenylalanine was produced after 6 h with 97% conversion rate. PMID- 12555548 TI - [Fraction P isolated from the ribosomes of Serratia odorifera and its tumor inhibiting effect]. AB - A protein(component P) with molecular weight of 16 kD was isolated by the method of preparation electrophoresis from the ribosomes of Serratia odorifera, which were obtained by the method of differential centrifugation. This protein showed specific immune response reactivity with the antiserum of the ribosomes mentioned above. Meanwhile, component P showed apparent protective function for QGY tumorcharged nude mouse and different ways of giving medicine. It was evident from study results that component P isolated from the ribosomes of Serratia odorifera showed a direct function for tumor growth inhibition and it was the effective component of the ribosomes of Serratia odorifera. PMID- 12555549 TI - [Studies on microbial production of undecane 1, 11-dicarboxylic acid from N tridecane]. AB - A mutant, Candida tropicalis P-12-242, which can produce undecane 1, 11 Dicarboxylic acid(DC13) from N-Tridecane(nC13), was obtained by treating the parent stain UH-2-48 with sodium nitrite. On 2500L fermenter testing, under the optimum condition where the fermentation medium contained total 26% nC13, pH of the course of fermentation was maintained range 7.5-8.0, at 28 degrees C-30 degrees C, the highest level of DC13 production was obtained after 6 d, and the average amount of DC13 accumulated was 182.6 g/L in broth. After received residual nC13, The average consumption rate of DC13 from nC13 was 87.8%. The purity of the product DC13, which was analyzed by gas chromatography was about 96.8%. PMID- 12555550 TI - [Advance of adaptive mutation research]. PMID- 12555551 TI - [The use of AFLP technique for the study of genetic diversity in peanut bradyrhizobia]. AB - A total of 146 bradyrhizobial strains including 133 peanut bradyrhizobia isolated from different regions and 13 reference strains representing B. japonicum and B. elkanii were studied for their genetic diversity by use of AFLP marker technique. The genomic similarity was analysed by numerical Taxonomy based on the AFLP patterns. The results showed that the genetic diversity of peanut bradyrhizbia were very high. Each strain had a identical AFLP pattern. AFLP technique is simple, fast, highly reproducible, and is a useful tool for evaluating genetic diversity. PMID- 12555552 TI - [Cloning of DNA fragment related to salt tolerance in Sinorhizobium meliloti 042B]. AB - Total DNA partially digested by EcoR I was prepared for S. meliloti 042B, in which 15-25 kb DNA fragments were collected. Vector pLAFR I was purified and digested by EcoR I, and then the various DNA fragments of 042B were ligated with pLAFR I by T4DNA ligase. Gene library of S. meliloti 042B was constructed with pLAFR I using E. coli S17-1 as recipient. The number of bacterial recombinants obtained was about 8,000 and 95% of them contained foreign DNA fragments. Using NTG, 042B was mutated on FY plates and 12 sensitive strains were screened at 0.5 mol/L NaCl from 2,000 colonies. One of them was named GZ17 and selected as a recipient strain. By biparental mating the foreign DNA fragments were introduced from gene library of strain 042B into recipient strain GZ17 which is sensitive to 0.5 mol/L NaCl. Then the transconjugants were grown on FY plates containing tetracycline (20 micrograms/mL) and 0.5 mol/L NaCl. A 7 kb inserted DNA fragment related to salt tolerance was obtained. In subcloning experiment, a 4 kb DNA fragment related to salt tolerance was obtained. PMID- 12555553 TI - [Cloning of mitomycin C resistance gene from Streptoverticillium caespitosum ATCC27422 and studying its function]. AB - Streptoverticillium caespitosum is the major producer of anti-cancer drug mitomycin. It demonstrated that there exists a strong restriction system for the transformation of foreign DNA in this strain. Using the blocked mutant S-6 which is sensitive to mitomycin C (MC) and does not produce mitomycin as a recepient and plasmid pIJ699 as vector, the gene library of the strain ATCC27422 was constructed. A clone of mitomycin C resistance gene(mcr) was obtained by screening from this library. The mitomycin C resistance gene cloned on the 6.2 kb fragment of plasmid pLX5 could be expressed in the Strptonmyces lividans TK54. It was the first time to introduce successfully the plasmid pLX5 into the wild type strain ATCC27422 by electroporation technique. And the transformant's resistance to mitomycin C has risen greatly: the MIC has gone up from 200 micrograms/ml to more than 1000 micrograms/ml. The result of fermentation showed that the yield of mitomycin of ATCC27422 harboring plasmid pLX5 was more than the wild strain ATCC27422's. It suggested that there is some relationship between the level of the resistance to mitomycin C and its biosynthesis. PMID- 12555554 TI - [Expression of xylose-metabolic key genes of Trichoderma reesei on various carbon sources measured by a series of Northern hybridizations]. AB - The expression of xylose reductase (XR), xylitol dehydrogenase (XDH) and transaldolase (TAL) genes from Trichoderma reesei, measured by Northern hybridization, were studied by adding different carbon sources (20 kinds, including single and mixed carbon sources) separately into the basal medium on which T. reesei QM9414 was grown. The experiment results indicated that the two disaccharides: sophrose and xylibiose act as a strong inducer for the expression of xr and xdh. The lactose and arabinose were identified as inducer also. The presence of glucose repressed the transcription of xr and xdh. When glucose depleted. xr and xdh were expressed at certain level, implying that expression of xr and xdh are controlled by the carbon catabolite repression mechanism and there existed a constitutive base level's expression of xr and xdh when the catabolite repression mechanism was derepressed. On the other hand, transaldolase gene strongly expressed on all the carbon sources used. PMID- 12555555 TI - [Transposition of Tn5096 in a agricultural antibiotic 120 roducer Streptomyces hygrospinocus var. beijingensis RF220]. AB - No transformant was obtained when pCZA168(bla, tsr, Tn5096, ColEI rep. Strep repts) was used to transform S. hygropinocus RF220. pIJ702 isolated from S. hygroscopicus N103 was transformed into RF220 at a low frequency. pIJ702 plasmid was cured in RF220 transformant and it was re-transformed into its cured FR220 strain, but the transformation frequency was not increased significantly, suggesting that restriction-modification system in FR220 was existent and complicated. Four transformants containing pCZA168 were obtained, when the RF220 strain was grown in medium with ampicillin, glycine and the protoplast was stored at -70 degrees C. Restriction analysis of plasmid from transformants indicated that the DNA fragment from E. coli in pCZA168 was deleted. With transposition of Tn5096, two mutants blocked in antibiotic biosynthesis of 120 and some mutants with variation in antibiotic level were obtained, this showed that the Tn5096 transposed in different positions of chromosomal DNA in RF220 and resulted in the production of 120 in different level. PMID- 12555556 TI - [New coleopterancidai strains of Bacillus thuringiensis and production of coleopterancide]. AB - Two coleopterancidal strains of Bacillus thuringiensis, YM-03 and SHQ11-10, were isolated from soil in China. H-Serotype of YM-03 was H8a8b, and H-Serotype of SHQ11-10 has not been identified. Both of them produced flat bipyramid-like crystals which consisted of 68-70 kD and 65 kD protein respectively. Bioassay indicated that these two Bt strains were very active on the larva of Leptinotarsa decemlineata and Plagioder aversicolora. They grew prosperously in fermentation. Powder of YM-03 product was high effect against colorado potato beetle in the field experiment. PMID- 12555557 TI - [The isolation and characterization of type 1 pili from pathogenic Escherichia coli of chicken origin]. AB - The pili from pathogenic Escherichia coli isolates 566, 1794 and TK3 of chicken and turkey origin were purified. After mechanic detachment from the bacterial cells, the pili were concentrated by precipitation with ammonium sulfate, dialyzed, and solubilized in buffer containing deoxycholate. The fraction containing the pilus was purified further by ultracentrifugation in a sucrose gradient. After ultracentrifugation, the pili at the density of 1.10 to 1.15 g.cm 3 (between 10%-20% of sucrose gradients) were collected, and the purified pili from strain 566, 1794 and TK3 had an apparent molecular weight of 17,500, 17,000 and 17,000 respectively, which retained their ability to bind the erythrocyte in a mannose-inhibitable fashion. Hyperimmunesera raised in BALB/C mice against the purified pili from strain 1794 reacted positively with type 1 pili from both isolates 566 and TK3 by immuno blot. These results revealed that the three strains either Chinese or north american isolates expressed type 1 pili which had molecular weights from 17,000 to 17,500, and they have common antigenic epitopes. PMID- 12555558 TI - [Distinction of yeast flocculent phenotypes and studies of the physiological and biochemical characteristics of yeast flocculation]. AB - More than 400 yeast strains were examined for their flocculation, and five of them displayed strong flocculation. The 5 strains were divided into two groups which are Flo 1 and NewFlo phenotypes on the basis of their response to sugar inhibition. Related physiological and biochemical characteristics of the two phenotypes' strains were studied. The results showed that the flocculation of Flo 1 phenotype was only inhibited by mannose; it was sensitive to high temperature (70 degrees C), pronase E, trypsin, whereas tolerant to pronase K, chymotrypsin, Ca2+, pH; the flocculation of NewFlo was inhibited by many sugars, such as glucose, maltose, sucrose, mannose; it was sensitive to high temperature (70 degrees C), pronase, Ca2+, pH. The Calcium concentration and pH value of the optimum flocculaton of the Flo1 and NewFlo phenotypes strains were respectively 10 mmol/L-1 mol/L; 3.0-4.5. PMID- 12555559 TI - [Stable expression and immunogenicity in a attenuated Salmonella typhi strain of coli surface antigen-6 of enterotoxigenic Escherichia coli]. AB - A gene fragment encoding for surface antigen CS6 of enterotoxigenic of Escherichia coli has been cloned into the plasmid pXL670, a new recombinant plasmid pSS64 was obtained by transforming E. coli X6097 with asd gene deletion. A safe and effective E. coli/S. typhi bivalent candidate vaccine was constructed by introducing pSS64 into delta aroA, delta aroC, delta asd Salmonella typhi. The vaccine strain is still stable in the absence of antibiotics. Animal tests demonstrated that this strain, when administered subcutaneously in mice, could provide significant protection against the intraperitoneal challenge from wild S. typhi Ty2. Immunization of rabbit with this strain raised specific antibody responses against CS6 and Vi antigen of S. typhi. This study lays the foundation for the construction of a new E. coli/S. typhi bivalent live oral vaccine. PMID- 12555560 TI - [Sugar phosphorylation activities in acetogenic bacteria]. AB - Seven acetogenic bacteria (Acetitomaculum ruminis, Acetobacterium woodii, Eubacterium limosum as well as isolates A2, A4, A10 and H3HH) were tested for PEP and ATP-dependent phosphorylation of glucose and 2-deoxyglucose. Although all organisms had detectable phosphorylation activity, substantial variation existed in the rates of both PEP- and ATP-dependent phosphorylation. Isolate Alo had the highest rate of PEP-dependent phosphorylation of 11.62 nmol.L-1.mg-1.min-1. Isolate A10, H3HH as well as E. limosum most likely have a glucose phosphotransferase system(PTS). In contrast, A ruminis, A. woodii and isolate A2, A4 had PEP-dependent glucose phosphorylation rates very similar to control rates, suggesting the lack of PTS activity. The rates of ATP-dependent glucose phosphorylation were higher than PEP-dependent phosphorylation in all organisms surveyed. However, substantial variation existed in the rates of ATP-dependent glucose phosphorylation. The glucose PTS of isolates A10 and H3HH were induced by the presence of extracellular glucose. Moreover, the specific activity of the glucose PTS of both isolates increased as cultures progressed from the early log to late log phase of growth. ATP- and PEP-dependent maltose and sucrose phosphorylation was detected in isolates A10 and H3HH. Although activity was detected in both isolates(A10 and H3HH), the rate of activity varied considerably, depending on the sugar and organism tested. PMID- 12555561 TI - [Culture conditions of engineered strain of L-asparaginase and the recombinant plasmid stability]. AB - The growth curves of engineered strain JM105(pASN) were different in LB and M-3 media. The expression level and activity of L-asparaginase were affected apparently by both biomass and induction time. Glucose repression of production of L-asparaginase was found. The stability of the recombinant plasmid pASN in different host strains and in LB and M-3 media was determined. After cultivation inLB broth and M-3 media at 30 degrees C for more than 50 generations without antibiotic selection, then induced at 42 degrees C for up to 5 h, the engineered strains were proved to be stable, except for DHA alpha (pASN). PMID- 12555562 TI - [Preparation of immobilized penicillin V acylase from Fusarium oxysporum and its properties]. AB - Extracellular penicillin V acylase from Fusarium oxysporum FP941 was partially purified by means of adsorption on, and elution from gamma-alumina and (HN4)2SO4 fractional precipitation. The enzyme was immobilized on acrylic polymer support linkaged with covalent bond. The activity of wet immobilized enzyme was 217 IU/g, The activity yield of Immobilized enzyme was 53%. Optimum pH and temperature for immobilized enzyme action wer 8.0 and 55 degrees C, respectively. The enzyme was stable below 50 degrees C and pH 4-11. The activity of the immobilized enzyme remained 90% after it was reused 25 times. PMID- 12555563 TI - [Molecular clone and sequence analysis of cDNA fragments of hog cholera virus strain C]. AB - The 5' nonencoding region, p23 and p14 encoding region and E1 gene of hog cholera virus (HCV) strain C were amplified from total RNA extracted from HCV strain C infected rabbit spleen by reverse transcription and nested or half--nested PCR. The PCR products were cloned into pGEM-T vector. Nucleotide sequencing was performed using an ABI PRISM sequencing device; based on the incorporation of fluoresect labelled dideoxynuclotide teminators. The obtained sequences on 5' noncoding region and part of p23 and p14 encoding region were compared with HCV strains Shimen and C sequenced in Moormann's lab. The result showed that the homology between HCV strains C sequenced in this report and in Moormann's lab was 99.19%, and the homology between HCV strains Shimen, the standard virulent HCV strain in China, and C sequenced in this report was 94.69%. It was also discovered that the base C at 244 of the genome of HCV strains Shimen and C sequenced in this report was absent at the genome of C strain sequenced in Moormann's lab et al. PMID- 12555564 TI - [Sequence analysis of the mutated AOX2 promoter region in the Pichia pastoris]. AB - Spontaneous revertants of the AOX1-defective P. pastoris strain have been isolated, which were identified as phenotypically utilize methanol to grow as the wild type. The promoter region of the AOX2 gene from the revertant has been obtained by PCR amplification, and the DNA fragment is 1022 base pair in size. By the analysis of sequencing result and compared with the AOX2 gene sequences recorded in Gene Bank, two point mutations which at positions of -529 and 255(relative to the translation initiation codon) respectively, have been found. Since the positions are located in the AOX2 upstream repression sequences, the mutations may act interfere the repressor to combine with the functional sequences, and increase transcriptional activity. The above result implicated that the P. pastoris' ability to utilize methanol could be increased through modification the upper stream transcriptional regulation region in the AOX2 gene. PMID- 12555565 TI - [Introduction and analysis of projects received and supported in division of microbiology NSFC in 1998]. PMID- 12555566 TI - [Study on polyphasic taxonomy of rhizobia isolated from Lespedeza species]. AB - The diversity of rhizobia isolated from Lespedeza spp. was determined on the basis of numerical analysis of phenotypic characteristics, sodium dodecyl sulfate polyacrylarmide gel electrophoresis (SDS-PAGE) of proteins, DNA-DNA homology and restriction fragment length polymorphism (RFLP) analysis of 16S-ribosomal DNA genes. According to numerical analysis of 125 phenotypic characteristics, strains were divided into two groups at a similarity level of 67%. Group I included all the fast-growing strains, group II included all the slow-growing strains. Above the similarity level of 80%, four subgroups could be further divided. Subgroup I was fast-growing rhizobia containing representative strain of Sinorhizobium saheli. Subgroup II, III, IV were slow-growing rhizobia. Subgroup II composed of strains isolated from Lespedeza cuneata in Beijing area and these isolates produced acid in medium containing mannitol. Subgroup III included type strain of Bradyrhizobium japonicum. The DNA G + C contents and DNA-DNA homology of the members of above four subgroups were determined. The subgroup I shared the same DNA homologous group with S. saheli, subgroup III belonged to B. japonicum, subgroup IV belonged to B. elkanii, subgroup II was an unique DNA homologous group which showed low level of DNA relatedness with other slow-growing rhizobia species. RFLP analysis of 16S rDNA genes verified that the subgroup II was a distinctive genealine and showed genetic variation within the strains in it. PMID- 12555567 TI - [Genetic diversity among Chinese peanut rhizobia by rep-PCR analysis]. AB - Repetitive sequences(repetitive extragenic palindromic[REP], and enterobacterial repetitive intergenic consensus [ERIC]) with the polymerase chain reaction(PCR) were used to fingerprint pure DNA extracted from 79 bradyrhizobial strains. These strains included 59 peanut rhizobia [Bradyrhizobium sp. (Arachis)] isolated from root nodules of 24 peanut (Arachis hypogaea L) cultivars from 23 sites in 11 provinces in China, 6 peanut rhizobial strains from other nations, and 14 reference strains of other bradyrhizobia. All the strains were clustered at the level of 11%, 2%, and 11, 24 clusters were obtained at the similarity of 50% with REP-PCR and ERIC-PCR fingerprints, respectively. The results showed that significant genomic diversity exists within the peanut rhizobia from China. Also they suggested that there are different distributions of REP and ERIC in genomic DNA of the peanut rhizobia. The combined results, REP plus ERIC data, were found between the both above. The genomic diversity seems not to correlate with their host and geographic origins. Our results supported this technique is a useful tool for genotypic characterization and identification of rhizobia as well as ecological studies. PMID- 12555568 TI - [Molecular cloning and sequencing of outer capsid protein gene of rice dwarf virus and its expression in Escherichia coli]. AB - The S2 full-length cDNA of rice dwarf phytoreovirus which enocodes the viral outer capsid protein was cloned and its complete nucleotide sequence was determined. The results showed that S2 is 3512 bp long with a large open reading frame which encodes a protein of 1116 amino acids. It shares 94.6% and 95.4% identity with RDV of Japanese H isolate in terms of nucleotide and amino acid sequences, respectively, and it also shows some homology with VP2 of rotavirus at the level of amino acid sequence. The search of deduced RDV S2 amino acid sequence in Blast network found that there were 4 leucine-rich motifs in P2 protein, and ten amino acids within the hydrophibic region at amino-terminus could form an alpha-helix. Predicted secondary structure of S2 cDNA indicated that a hairpin and a stem loop are present in the 5'-end within 50 bp, and a stem loop in the 3'-end within 50 bp. RDV S2 partial and full-length sequences were then cloned into expression vector pET-11d & pTrcHisC. SDS-PAGE and Western blot proved that amino- and carborn-termini of P2 were successfully expressed in E. coli. PMID- 12555569 TI - [Amplification of phytoplasma 16S rDNA from banana bunchy top disease, RFLP and sequence analysis of 16S rDNA fragment]. AB - Extract DNA from banana plants with banana bunchy top disease and amplify the phytoplasma 16S rDNA fragment by nested PCR. The classification of the phytoplasma established on the basis of 16S rDNA restriction patterns was examined by performing a sequence analysis of 16S rDNA fragment. The RFLP pattern, sequence data and phylogenic tree showed that this phytoplasma should belong to Group I(Phytoplasma asteri). PMID- 12555570 TI - [Cloning of a gene encoding cytoplasmic glycerol-3-phosphate dehydrogenase from Candida glycerolgenesis]. AB - The response of the yeast Saccharomyces cerevisiae to osmotic stress is to synthesis and accumulate the glycerol in order to increase the internal osmolarity and this response is controlled by the high-osmolarity glycerol (HOG) response pathway, whose important target gene is GPD1. The increase of the activity of glycerol-3-phosphate dehydrogenase by over-expression of GPD1 gene can increase the glycerol yield greatly. In this study, a gene encoding cytoplasmic glycerol-3-phosphate dehydrogenase of Candida glycerolgenesis was cloned out by inserting Sau3AI-generated chromosomal DNA fragments into the BamHI site of a yeast-E. coli shuttle vector, YEp51. Fifteen transformants were isolated on a supplemented minimal medium containing 50 g/L of sodium chloride from the constructed C. glycerol-genesis genomic library by using genetic complement approach. The recombinant plasmid, YEp0601, from transformant 0601, possessed the genetic markers of YEp51 and was able to restore the osmotolerance of S. cerevisiae 642(gpd1 delta, gpd2 delta). These indicated that a gene coding for cytoplasmic glycerol-3-phosphate dehydrogenase of C. glycerolgenesis was successfully cloned out. PMID- 12555571 TI - [Nucleotide sequence analysis of the minimal replicon of the Streptomyces plasmid pSGL1]. AB - The high-copy-number plasmid pSGL1(7.4 kb) was isolated from Streptomyces globisporus. Deletion experiments showed its minimal replicon is located on the 2.0 kb Sau3 AI fragment. This fragment was subcloned. DNA sequence data analysis showed this fragment is a new sequence. Only an open reading frame with high coding probability is located on the minimal replicon. The deduced protein contains motifs characteristic of replicase for rolling-circle replication. PMID- 12555572 TI - [Expression, purification and bio-activity analysis of oncostatin in GST-fusion expression system]. AB - Oncostatin (OSM) is an important cytokine which has diverse biological activities. It may be useful in basic scientific and clinical studies. OSM was efficiently expressed by cloning the OSM gene into GST fusion expression vector. The level of expression of the fusion protein was about 50% of the total cell protein by SDS-PAGE analysis. The percentage of fusion protein in soluble protein could be 15% when the induction was in low temperature. The purity of purified GST-OSM with an affinity chromatography column was about 90% after the inclusion bodies had denatured and renatured. The result of activity study showed that the first two amino acids in N-terminal of the OSM were important to the biological activity of OSM. PMID- 12555573 TI - [Purification and partial characterization of antagonistic proteins from Bacillus subtilis B034]. AB - Bacillus subtilis B034, an antagonistic bacteria isolated from the phyllosphere of rice, strongly inhibits the growth of Xanthomonas oryzae pv. oryzae which causes the rice bacterial blight. Crude extract was obtained by precipitation of the cell-free culture of B034 with 70% saturated (NH4)2SO4. The suspension of the precipitate strongly inhibits Xanthomonas oryzae pv. o-ryzae, and it is thermostable, resistant to trypsin, partial sensitive to proteinase K, pronase E and chloroform. Its active pH range is wide, from 4.0 to more than 12.0, relative more stable in high pH. Two antagonistic peaks were obtained from the crude extract of B034 after Phenyl-Sepharose CL-4B, DEAE-Sephacel and FPLC Superdex 75 HR 10/30 column chromatography. One of these two peaks, named P2, was showed only single band with 50.3 kD and pI6.25 in SDS-PAGE and PAGE-IEF, respectively. The N terminal partial sequence of protein P2 was analyzed. PMID- 12555574 TI - [Purification and characterization of a novel antifungal peptide APS-1 produced by Bacillus cereus]. AB - In previous study, we isolated an antagonist Bacillus cereus strain: S-1 from cotton plant. In field experiments, this bacterium was shown strong inhibition to several plant diseases. In this paper, we reported the purification of the antifungal substance produced by the bacterium and its properties. After the steps of acid precipitation, methanol and ethy lether extraction, Sephadex G100 and DEAE52 column chromatography, the antifungal material was purified. The purified material had absorption peak at 275 nm, and was exhibited negative in biuret color reaction. However after hydrolyzed with HCl, this substance shown positive in the same reaction. Amino acid analysis to the hydrolysate of APS-1 showed that APS-1 was composed of Glu, Asp, Tyr, Ser, Thr, Pro, Leu, Ile, Val and an unknown amino acid. Combining with its pratial resistance to proteinases, it was suggested that this antifungal material was a cyclic peptide. This peptide, named APS-1, with strong inhibition on the germination of spores of the phytopathogens tested, was shown high stability against ultraviolet radiation and heat. APS-1 may have potential role in plant diseases biological control. PMID- 12555575 TI - [Screening of acidic xylanase producing strain and studies on its enzyme production conditions]. AB - From 150 fungal strains, the authors found 8 strains contained mainly of xylanase activity over 100 U/mL in which the No. 149 strain was the highest xylanase producer. Which tentatively identified as Aspergillas niger. The appropriate medium composition was as follows: wheat bran hemicellulose 4%; NaNO3 1%; wheat bran 1% prepared in Mandels nutritional solution without (NH4)2SO4 and urea. After cultivated in shake-flask at 28 degrees C-32 degrees C for 60 h, the activity reached the highest value of 357.2 U/mL. The optimum pH of xylanase was 4.6 and it was stable at pH3-11. The fermented broth of strain 149 contained in addition to xylanase (relative activity 100) also included amylase(1.8), mannanase(0.98), beta-xylosidase(0.94) and cellulase(0.17). PMID- 12555576 TI - [Fermentation conditions for production of glutathione by recombinant Escherichia coli]. AB - The fermentation conditions for production of glutathione by recombinant E. coli were investigated, the addition of certain materials, such as yeast extract, precusor amino acids and ATP were mainly focused on. The results showed that the addition of precusor amino acids and ATP could promote intracellular GSH accumulation. DCW and intracellular GSH content would be 24% and 1.4 times higher than that of fermentation without additions when 2.0 g/L ATP and 9 mmol/L precusor amino acids were added at the begining and 12 h of fermentation, respectively. By using the optimized combination of additives obtained from orthogonal experiments, the maximal DCW and total GSH in broth could be improved 10% and 26% higher than the best results in orthogonal experiments, respectively. Based on the analysis of glucose utilizing ability of this strain, an exponential fed-batch culture process was conducted. DCW and total GSH in broth culd be 8.3 and 4.6 times higher than that of shaking flask culture and finally reached 80 g/L and 880 mg/L, respectively. PMID- 12555577 TI - [The effect of N+ implantation on SOD activity of Deinococcus radiodurans and induction of Mn-SOD]. AB - The induction of Mn-SOD and effect of N+ of 20 keV implantation on SOD activity of D. radiodurans were investigated. The results were as follows: (1) SOD activity of D. radiodurans had a little change when implantation dose was less than 8 x 10(14) N+/cm2, and increased gradually from 8 x 10(14) to 60 x 10(14) N+/cm2, and then decreased rapidly; (2) the test of adding H2O2 and CHCl3 CH3CH2OH which could inhibit different types of SOD showed that most SOD activity of D. radiodurans was constituted by Fe-SOD in general physiology and less then 8 x 10(14)N+/cm2 dose, and higher implantation dose induced increase of Mn-SOD activity. PMID- 12555578 TI - [Industrial scale-up of continuous ethanol fermentation using yeast flocs]. AB - Inducing yeast cells to self-flocculate as a cell immobilization method, an industrial scale-up plant composed of 4 air-lift suspended-bed bioreactors in parallel and with a total volume of 400 m3 was established and operated. During a more than 6 months continuous operation, the effluent contained ethanol of 70-80 g/L and residual sugar less than 5 g/L and the ethanol productivity of 7-8 g/L.h 1 was achieved. PMID- 12555579 TI - [Effect of promoter-PTH4 on Streptomyces sporulation]. AB - 233 bp of promoter--PTH4 was subcloned into Streptomyces plasmid pIJ4083. pIJ4470 was obtained after recombinant plasmid was introduced into Streptomyces coelicolor J1501. Both Streptomyces coelicolor J1501/pIJ4083 and J1501/pIJ4470 were grown on minimal medium (MM) agar containing mannitol and 10 micrograms/mL thiostrepton for 6 days, and then ultrathin sections of colonies were prepared and stained with silver proteinate to observe glycogen biosynthesis in cell by the electron microscope. Result showed that the glycogen was produced by J1501/pIJ4083, whereas little glycogen was produced by J1501/pIJ4471. The dark color spores were not observed in J1501/pIJ4470 with lead stain in contrast with dark color spores which were produced by J1501/pIJ4083, indicating that promoter PTH4 may play an important role in physiological and morphological differentiation of Streptomyces. PMID- 12555580 TI - [Conjugational actinomycetes of plasmid RSF101 from Escherichia coli to the rare actinomyceces of Nocardia asteriodes and Streptoverticillum caespitosus]. AB - RSF1010 is a naturally occurring Escherichia coli broad host-range plasmid about 8.7 kb in size. It can be mobilized at high frequency between different gram negative bacterial species when transfer functions are available in trans. Following the pioneering work of conjugational transfer of RSF1010 from E. coli to Streptomyces lividans and Mycobacterium smegmatis, the transfer of this plasmid by conjugation from E. coli S17.1 tp two gram-positive rare actinomycetes, Nocardia asteroides 3927 and Streptoverticillum caespitosus ATCC27422 was first time reported in this study. Southern blot analysis of the total DNA extracted from the actinomycetes' exconjugants proved that RSF1010 had been transferred from E. coli into the two new hosts and maintained staby in the exconjugants. Meanwhile, partial deletions of RSF1010 replicon loosing its antibiotics resistance makers were readily detected in E. coli. The implenmentation of this observation was discussed. PMID- 12555581 TI - [Detecting microorganisms in soil by PCR and molecular hybridization]. AB - PCR and molecular hybridization provide new methods to detect microorganisms in environment. A simple, sensitive protocol was developed to detect Bacillus thuringiensis HD-1 in soil by PCR and Southern Blotting in this study. Methods of extracting DNA from soil and purification were compared. By a modified total DNA extraction from soil and purification, followed by PCR and Southern Blotting, strain HD-1 released into soil can be detected with a sensitivity of 2.2 x 10(2) cells/g soil. By using other specific primers, this detection protocol can be widely used in the research of ecology and risk assessment of genetically engineered microorganisms released into environment. PMID- 12555582 TI - [Review and suggestion on science funds on microbiology in NSFC]. PMID- 12555583 TI - [Contribution of the transesophageal electrophysiologic study to the etiologic diagnosis of wide QRS tachycardia]. AB - INTRODUCTION: A wide-QRS complex tachycardia is suggestive of a ventricular tachycardia (VT). Its diagnosis requires an intracardiac electrophysiological study. That study is sometimes difficult to indicate in old or very young patients. The purpose of the study was to evaluate the interest of a rapid and noninvasive study by transesophageal route for the evaluation of the nature of a wide-QRS complex tachycardia. PATIENTS AND METHODS: Forty patients, aged from 16 to 85 years, without bundle branch block (BBB) in sinus rhythm, were admitted for documented wide-QRS tachycardia. Transesophageal electrophysiologic study (EPS) using one and two extrastimuli was performed in control state and after infusion of 20/30 micrograms of isoproterenol. Intracardiac EPS was performed in a second time in 38 of them. RESULTS: The study was negative six patients; intracardiac EPS remained negative in four of them, induced a VT in one and a Mahaim-reentrant supraventricular tachycardia in another one. Clinical tachycardia was induced in remaining patients: in 27 of them, the diagnosis of SVT with aberrancy was assessed; in other patients, the diagnosis of VT was assessed; The VT was a verapamil-sensitive VT or a bundle branch reentry (n = 7). The diagnosis was confirmed by intracardiac study. CONCLUSION: Esophageal EPS was a means to reproduce the clinical tachycardia in 34 of 40 patients and to evaluate the mechanism of wide-QRS tachycardia in 33 of 34 patients; this technique easy to perform should be indicated in patients in whom intracardiac study is debatable to avoid to diagnose by excess a VT or in the opposite to miss this diagnosis. PMID- 12555584 TI - [Beating heart surgery and left ventricular dysfunction]. AB - We report our experience with systematic coronary revascularization on the beating heart among patients with left ventricular dysfunction as defined by a left ventricular ejection fraction < or = 40%. Between September 1996 and April 2000, 500 off-pump (OPCAB) revascularizations were performed (95% of all revascularizations for the same time frame, single surgeon). Among them, 76 patients qualified as left ventricular dysfunction and were compared to a similar cohort of 237 patients operated on with cardiopulmonary bypass (CPB) during the same time frame. Age and sex distribution, average preoperative left ventricular ejection fraction and incidence of preoperative unstable angina were the same for both groups. On average, 3.04 +/- 0.89 and 2.97 +/- 0.69 grafts/patient were made in the OPCAB and CPB groups respectively (p = NS). Complete revascularization was achieved in 95% of the OPCAB group. Incidence of preoperative intra-aortic balloon assistance were higher in OPCAB (22% versus 9%, p = 0.005) whereas postoperative need for new intra-aortic balloon assistance was higher in CPB (8% versus 0%, p = 0.02). Incidence of postoperative myocardial infarction was comparable in both groups (6.5% versus 5.5%). Maximal creatinine phosphate of myocardial origin were lower in OPCAB group (beating heart: 32 +/- 52%, cardiopulmonary bypass: 45 +/- 51%, p = 0.055). Operative mortality was lower in OPCAB group although it did not reach statistical significance (beating heart: 2.6% versus cardiopulmonary bypass: 4.6%, p = 0.3). Complete coronary revascularization on the beating heart can be achieved in patients with left ventricular dysfunction with excellent outcome and low operative mortality. PMID- 12555585 TI - [Role of surgery in subaortic stenosis. Report of 56 cases]. AB - INTRODUCTION: Subaortic stenosis is a rare congenital heart disease defined as a left ventricular outflow tract obstruction. We reviewed our surgical experience in this cardiac disease with particularly attention to the different anatomical types of the obstruction. PATIENTS AND METHODS: From January 1987 to December 1998, 56 patients with a mean age of 12.4 years underwent surgical treatment of subaortic stenosis in our Institution. The diagnosis included: subaortic membrane in 44 cases, fibromuscular process in seven and tunnel like hypertrophy in five. RESULTS: There were two hospital deaths (2/56 = 3.5%) and three patients presented postoperative heart block. The first postoperative echocardiographic control showed a mean fall in left ventricleaorta gradient of 78%. In a mean follow-up of 36 months, there were no deaths. All patients periodically controlled, showed an echocardiographic progression of the gradient and it was not related to the different anatomical types of the obstruction. There were no signs of aortic insufficiency progression. CONCLUSION: We can affirm that the surgical treatment of the subaortic obstruction is simple and safe. The medium and long-term progression toward the recurrence is independent to the anatomical type and justify the need of serial echocardiographic control. PMID- 12555586 TI - [Surgery of active-phase infective endocarditis: report of 13 cases]. AB - OBJECTIVE: Infective endocarditis is always a disease at the present time. In this work we report our initial experience of infective endocarditis surgical treatment during the acute phase. METHODS AND RESULTS: Between September 1993 and December 1997, 13 patients underwent valvular surgery for native infective endocarditis. Mean age was 31 +/- 11 years (range: 9 to 42 years); 92.3% of the patients presented with pre-existing rheumatical valvular lesion. Surgical indication was the occurrence of heart failure in all these patients. Surgery consisted on aortic valvular replacement in four patients, mitral and aortic valvular replacement in nine patients. Tricuspid valvuloplasty was required in two patients. Early mortality was 15% (two patients). A third patient developed cardiac failure and died three years postoperatively. CONCLUSION: This work emphasises the interest of the surgical treatment in the active phase of the infective endocarditis. PMID- 12555587 TI - [Anti-digoxin antibodies in severe digitalis poisoning in an 11-day old infant. Review of the literature]. AB - Acute digitalis intoxication bringing into play the prognosis for survival is a rare complication in children, particularly in the neonatal period. We describe the case of an accidental massive digitalis intoxication in a newborn aged 11 days, which caused a complete atrioventricular block and a state of cardiogenic shock. The outcome was favorable four hours after administration of the first dose of anti-digoxin antibodies, with a complete reversal of clinical and electrocardiographic signs. The onset of first-degree atrioventicular block 48 hours afterward made us consider the possibility of the tissular salting-out effect of the digoxin and led us to administer a second dose of specific antibodies. The originality of this case has to do with the severity of the initial clinical picture, its total reversal with antibodies and the salting-out phenomena that followed. The case reminds us, along with the data in the literature, the criteria of wrong prognoses in massive digitalis intoxication in the child and the indications for anti-digoxin antibodies. PMID- 12555588 TI - [Anomalous origin of the left coronary artery from the pulmonary artery: report of 4 cases]. AB - Ahomalous origin of the left coronary artery from the pulmonary artery is a rare but severe congenital cardiopathy that often causes myocardial infraction within the first months of birth, often with congestive heart failure. We report four cases, three boys and one girl. The appearance of the first symptoms is early, usually between the second and the fifth month. The chest X-ray showed cardiomegaly. Echocardiography showed dilated and hypokinesis cardiomyopathy. In all patients the diagnosis was suspected because the standard electrocardiogram showed a constant deep Q wave in leads I and aVL. Angiography with coronarography was realized in only two cases confirming the diagnosis. The left coronary was directly reimplanted into the aorta in one child at 12 months, he has improved throughout six years. Two infants died after four and six years; one child had a spontaneous progressive recovered normal ventricular function because he had an important collateral circulation from right coronary artery. In conclusion, this rare ischemic myocardiopathy is serious because it's life threatening. The indirect signs given by the electrocardiogram and the echocardiography are of a big diagnosis aid. The prognosis depends on the precocity of the chirurgical reimplantation. PMID- 12555589 TI - [Cardiovascular effects linked to the use of chloroquine]. AB - The chloroquine is the mainly and most frequently drug used as antimalaric in the world, in spite of the extension of resistance phenomena. Besides, the chloroquine is also commonly indicated in rheumatology and dermatology as a chronic treatment of some connective tissue disease. The chloroquine has three main cardiovascular effect: membrane stabilizer, direct negative inotropic effect and direct arterial vasodilator. Thus, these cardiovascular iatrogenic effects of the chloroquine are important both through their potential frequency and seriousness. Personal clinical cases and medical review enables to identify the main effects, observed either with prophylactic, or curative, or even toxic dosages. The more often, there are some rhythm and conductance disorders, myocardiopathy, even sometimes vasoplegic shocks. A list of the commercial patent medicine including chloroquine enables to be aware, to prevent and to take into account the cardiovascular risks of a treatment newly set or carried on for long years. PMID- 12555590 TI - [Cardiovascular effects of doping]. AB - Cardiovascular effects of doping drugs are numerous, with different mechanisms: vasoconstriction of amphetamines, erythropoietin and cocaine; sodium water retention of anabolic steroids and corticosteroids; elevation in blood viscosity of erythropoietin, perflurocarbon emulsion, recombinant hemoglobin and anabolic steroids; sympathetic nervous system activation of amphetamines, beta 2 agonists and clenbuterol; lipids profile disorder of anabolic steroids. Physical activity consequences, particularly bradycardia and dehydration, are worsening. Thrombosis and arrythmogenic effects, with possibility of sudden death, are the severe immediate events. Hypertension and coronary diseases are medium-term effects; acute myocardial infarction is frequent. Heart failure can be secondary to cardiac muscle direct fibrosis, like with anabolic steroids. These cardiovascular effects are serious and it is necessary to early detect the doping drugs use in sporstmen; all prescribing physician should be aware of existing drugs and their clinical events. PMID- 12555591 TI - [Medico-economic impact of the Program of Medicalization of the Information System on hospital stay for myocardial infarction: influence of the quality of data collection and hospitalization length]. AB - The aim of the Medicalisation Program of the Information System was to describe the activity of hospital for budget allocation. This work concerned the whole hospitalizations in the unit of intensive care of cardiology of Dijon for a myocardial infarction (MI) during the 1st half of 1998 (59 patients). The objectives of this study were: 1) the estimate of the real cost of MI management; 2) the comparison of this cost with the reference cost, determined from the data of the National Basis of Costs (BNC); 3) the economic impact of the quality of coding. The real global cost of MI was estimated at 2,323,542 FF (average by patient: 39,382 +/- 15,718 FF). Sixty eight per cent of the costs are directly related to the standing fixed overheads; in contrast, the medical and the therapeutic acts accounted for only 32% of the estimated real cost. A 52% over estimation was highlighted between the estimated real cost and the cost of reference (p < 0.001). The errors of coding accounted for an under-estimation of only 3.6% of the cost of reference. The duration of hospitalization was significantly higher than the stay length taken from the national reference database (12.9 +/- 5.4 versus 9.2 +/- 2.1 days; p < 0.001), and was mainly responsible for these discrepancies of costs. PMID- 12555592 TI - [Left ventricular hypertrophy in hypertensive African blacks: echocardiographic study of 452 patients]. AB - OBJECTIVE: The aim of this study was to define in a population of hypertensive Black African, the frequency of left ventricular hypertrophy (LVH), the different kind of LVH, and the factors of LVH occurring. METHODS: Clinical and echographic (according to Penn convention) data was collected in 452 consecutive hypertensive patients (193 women and 259 men). LVH was defined as an indexed left ventricular mass (ILVM) > 134 g/m2 in man and 110 g/m2 in woman. RESULTS: The mean systolic blood pressure was 187 mmHg, and the mean diastolic, 111 mmHg. The mean ILVM was 135.25 g/m2. LVH was present in 53.3% of the patients. Concentric LVH represented 48.5%, eccentric LVH 41.1%, and asymmetric hypertrophy of the septum, 10.4%. LVH increased with patient's age (p = 0.008), hypertension severity (p = 0.0001), and the ancientness of the hypertension (p = 0.0001). CONCLUSION: LVH is frequent in hypertensive black African, and concentric LVH is the most frequent kind of LVH. Factors of LVH occurring are patient's age, severity and the ancientness of the hypertension. PMID- 12555593 TI - [Transformation of asymptomatic Wolff-Parkinson-White into a symptomatic form as a result of the ablation of the bundle of Kent]. AB - We report the case of an asymptomatic young man, 19 years old, who had a Wolff Parkinson-White syndrome. A form at risk of potentially severe arrhythmias was detected by the electrophysiological study. The Kent bundle radiofrequency ablation was performed with success. However the patient developed few weeks after, spontaneous paroxysmal junctional tachycardias. Such report indicates that we should remain careful in asymptomatic patients. PMID- 12555594 TI - [Hydatid cyst of the heart simulating coronary ischemia]. AB - Cardiac echinococcosis is rare despite endemic occurrence of echinococcosis in some regions of the world. Clinical presentation can vary and may be misleading. Diagnosis is facilitated by new imaging techniques. Surgery is mandatory to avoid serious complications. We report a case of cardiac echinococcosis with pseudo, ischemic clinical and electrical presentation. PMID- 12555595 TI - [Aneurysm of the posterior sinus of Valsalva ruptured into right atrium]. AB - Sinus of Valsalva aneurysm is rare malformation, which takes origin from one of the three aortic sinuses. Diagnosis of this abnormality is usually made when rupture has occurred. Our observation underlines the severity of the posterior sinus of Valsalva aneurysm ruptured into right atrium, and emphasizes the utility of transthoracic and transoesophageal echocardiography for diagnosis. PMID- 12555596 TI - [Invasive thymoma extending into the superior vena cava and the right atrium: a case report and review of the literature]. AB - If the neoplastic extension with the higher vena cava can be observed during the evolution of the invasive thymomes, the tumoral extension in the right auricle, remains on the other hand an exception. We report an invasive case of thymome to higher intracellar extension, right intraatrial and pleuropulmonary revealed by a syndrome undermines higher and a left pleurisy. The transthoracic echocardiography supplemented by the echocardiography transoesophageal pose the cardiac diagnosis of tumor, and it is the thoracic tomodensitometry which highlights a tumoral process mediastinal of malignant pace invading the left inominal venous trunk, the higher vena cava and extending in the right auricle. The diagnosis of certainty is carried by the endobronchial biopsy and the puncture tomodensitometric biopsy under control of the mediastinal mass after anatomopathological examination. Under chemotherapy, the evolution over 18 months is marked by the absence of cardiovascular complications in spite of the non regression of the tumoral mass. This observation stresses the importance of the realization of the echocardiography especially transoesophageal in front of all invasive thymome and the reliability of the puncture biopsy scanoguided like diagnoses technique not very invasive in the forms not extirpables. The forecast of this affection depends on the effectiveness of the processing. The surgery when it is possible, remains the principal therapeutic measurement which really proved reliable. PMID- 12555597 TI - [Short- and long-term benefits of smoking cessation]. AB - The beneficial effects of withdrawal smoking on the disrupted physiological functions bound to tobacco spread differently: some weeks for smell, taste and breathing, to the less six months to recover tolerance to the effort without dyspnea. For the cardiac attack the risk bound to tobacco disappears after five years, and the risk for cancers after ten years. The expectation of life of the ex-smoker joined the one of non-smokers 15 years after the stop of smoking. The tobacco use by the women raise some particular problems. To convince the smoker to persevere in the decision of withdrawal smoking and to be free of dependences (pharmacological and psychological) it could be necessary to use the nicotinic substitutes and/or a psychological support. Stop smoking is the better and the most economic therapeutic to avoid recurrence of cardiac attack for a lot of authors. PMID- 12555598 TI - [Deleterious cardiac effects of serotonin in myocardial ischemia: role of naftidrofuryl]. AB - It is now accepted that serotonin can either initiate or aggravate myocardial ischaemia through a vasoconstrictor action and platelet activation. It is therefore possible that substances likely to neutralize the effects of serotonin could be used, without any danger, in humans with ischaemic heart disease. This type of action may therefore be exerted by 5-HT2 antagonists, such as naftidrofuryl. A recent double-blind clinical study has in fact shown that administration of naftidrofuryl versus placebo leads to better exercise tolerance, with an increase in the maximum level and delay in ST segment shift (increase in the threshold of onset of ischaemia). The purpose of this study was therefore to evaluate, in an animal model (pig) of acute myocardial ischaemia (occlusion of the proximal section of the left anterior descending coronary artery), the action of serotonin, naftidrofuryl and a combination of both substances on the following parameters: 1) electrophysiological (sinus heart rate, ST segment shift, T-wave amplitude, duration of monophasic action potentials, intraventricular conduction time); 2) haemodynamic (systolic, diastolic and mean blood pressure, first derivative of rate of increase of left ventricular pressure with time: LV dP/dt max); and 3) biochemical (malonedialdehyde concentration as an index, cell peroxidation index, creatine phosphate and adenosine triphosphate). It was found that co-infusion of serotonin aggravated the myocardial ischemia and that naftidrofuryl exerted beneficial effects on the serotonin-mediated aggravation of myocardial ischaemia. PMID- 12555599 TI - [Vacation with a different ending]. PMID- 12555600 TI - [Expectations and obligations]. PMID- 12555601 TI - [Disease-mongers]. PMID- 12555602 TI - [Asylum and persons seeking asylum]. PMID- 12555603 TI - [Norwegian use of Internet health services]. AB - BACKGROUND: This study investigates the assumption that the Norwegian population's use of Internet health services has an impact on health related attitudes and behaviour. MATERIAL AND METHODS: In 2000 and 2001, 1,006 and 1,018 subjects were randomly selected from the general population in Norway and interviewed over the telephone. In 2001, a web-based questionnaire was presented to 600 of a 2,800 member Internet panel who reported having used the Internet for health purposes; 79% responded. RESULTS: The proportion of Norwegians who use the Internet for health purposes increased from 19% in 2000 to 31% in 2001. The proportion of those wishing to use e-mail in interaction with their doctor increased from 30% to 45%. Based on information from the Internet, 33% of users in 2001 have asked their doctor specific questions; 11% have suggested a diagnosis, 21% have altered their diet and/or lifestyle; 10% have started with health products or programmes without consulting their doctor. 13% have experienced anxiety, while 48% have experienced relief. INTERPRETATION: Norwegians' use of the Internet appears to supplement rather than replace ordinary health services. Health personnel will need to respond to Internet generated expectations and behaviour. PMID- 12555604 TI - [Paper or screen, mother tongue or English--which is better?]. AB - BACKGROUND: The trends in medical publishing are towards electronic versions and the use of the English language. The objective of this study was to compare general practitioners' ability to retain information when reading on paper versus on screen, and in their mother tongue versus in the English. MATERIAL AND METHODS: Randomized controlled trial of 114 Scandinavian general practitioners. They read a review article for 10 minutes either on paper/in English, on screen/in English, on paper/in their mother tongue, or on screen/in their mother tongue. Afterwards they completed a questionnaire with six open questions from the article. The main outcome measure was sum score on a scale from 0 (no correct answers) to 13 (all questions answered correctly). RESULTS: We found no difference between readers of paper and screen versions (median (interquartile range) 4 (2-6) vs. 4 (2-5), p = 0.97). Those who read their mother tongue scored significantly higher than those who read English (4 (3-6) vs. 3 (2-4), p = 0.01). INTERPRETATION: The medium (paper vs. screen) does not influence the ability of general practitioners to retain medical information. Doctors best retain medical information when reading it in their mother tongue. PMID- 12555605 TI - [Neurologic decompression sickness in sports divers]. AB - BACKGROUND: Sports diving is a popular recreational activity. Sports divers presenting with acute decompression sickness may exhibit residual neurologic and neuropsychological symptoms during follow-up, in spite of appropriate treatment. MATERIAL AND METHODS: A retrospective review of medical records was carried out for sports divers admitted to the department of neurology at Haukeland University Hospital during 1997. RESULTS: 11 out of 20 divers experienced residual neurological symptoms after treatment. Five responded poorly to treatment, with 50% or more residual clinical score. These patients tended to be older, had performed deeper dives, and more repetitive diving. Seven divers had increased slow wave activity in EEG on initial recording, in two the EEG changes persisted after treatment. INTERPRETATION: In this small series of sports divers with decompression sickness and arterial gas embolism, most neurologic symptoms responded to hyperbaric treatment. However, more than one half of the divers had residual neurological symptoms on discharge. Sensory loss and asymmetrical reflexes were the most common residual findings. PMID- 12555606 TI - [The corvette "Nordstjernen's" voyage to the opening of the Suez Canal--naval medicine in 1869]. AB - When the Norwegian corvette Nordstjernen was in the North Sea bound for Port Said to be present at the opening of the Suez Canal on 17 November 1869, an officer suffered a rupture of m. triceps brachii when he was drawn into the machinery during a storm. He was put ashore in Harwich; four days after the injury he was hospitalized in Colchester. The voyage was eventful in other ways too. Another officer died from typhoid fever in Ismailia. On the Swedish frigate Vanadis, also present at the opening of the Suez Canal, one of the doctors died from lung infection and was buried in Smyrna; a twelve-feet high column of white marble was taken from the ruins of Aesculap's temple and put on his grave. Denmark was represented by the frigate Sjaelland. During a storm in the North Sea, one seaman fell down on the deck from the foresail yard and suffered contusions and a fracture of the left clavicle. These cases illustrate challenges that faced our ancestors. The accident happened when the ship was in the Netherlands sector of the North Sea as we know it today. Today the Coast Guard could have arranged transport by helicopter and hospitalized the patient in about two hours. PMID- 12555607 TI - [Was it a sin? Emil Aarestrup, physician and poet]. AB - Emil Aarestrup (1800-56) worked as a general physician, later as a public health officer in Denmark. He married his cousin and wrote love poems to her and to other women. His erotic poetry is regarded as the most elegant in Danish literature. His poems have precise words and colourful details and show a special sense of the condensed mood in the erotic situation. He writes upon the complex nature of love: union, separation and death. PMID- 12555608 TI - [Serotonin syndrome--several cases of this often overlooked diagnosis]. AB - During the recent decade an increasing number of inquiries concerning cases of overdoses exhibiting typical signs of the serotonin syndrome have been recorded at the Swedish Poisons Information Centre. Four of these cases are presented together with a review of the literature. All patients had overdosed moclobemide and in one case this was the only drug taken. The other patients had ingested moclobemide together with citalopram (2 cases) and clomipramine (1 case). Moreover, other serotoninergic pharmaceuticals as sertraline and sumatriptan were simultaneously ingested in one case and buspirone in another. Three of the cases had hyperthermia, > 40 degrees C and the same number showed pronounced muscle rigidity, coma and mydriasis. Other severe signs and symptoms upon admission included positive Babinski and trismus in two cases each and seizures in one. All patients received mechanical ventilation. Two were treated with dantrolene sodium and one of them was given cyproheptadine as well. One patient received cyproheptadine treatment alone and another prolonged muscle relaxation. Three patients had a typical short clinical course, whereas one patient developed rhabdomyolysis, DIC and arrhythmias. All patients fully recovered. PMID- 12555609 TI - [Plants and plant produce--about plants as cause of diseases]. AB - BACKGROUND: Plants and plant products may lead to a variety of skin disorders, commonly referred to as phytodermatoses. MATERIAL AND METHODS: On the basis of literature studies and on the author's own experience, this article discusses the main groups of phytodermatoses, focusing on Norwegian plants that may cause skin disorders. RESULTS: A small number of plant families account for more than 95% of the phytodermatoses. Contact allergy to plants of the Compositae family is an important cause of skin disorder. Phytodermatoses are commonly related to the patient's occupation. INTERPRETATION: Doctors should be familiar with the term phytodermatoses and be aware of plants and plant products as possible causes of skin disorder. PMID- 12555610 TI - ["New wonder pill!"--what do Norwegian newspapers write]. AB - BACKGROUND: The news media are an important source of information on new medical treatments. There is, however growing concern that some of the coverage may be inaccurate and overly enthusiastic, thereby misleading the general population. METHODS: Eight major Norwegian newspapers were screened for articles on the benefits and risks of all the 60 new medications introduced on the Norwegian market from July 1998 through March 2000. We found a total of 492 articles describing 18 new medications over the period January 1998 through June 2000. RESULTS: Of the 357 stories that mentioned benefits, 79% did not report this in any greater detail, 51% gave the medication a positive coverage, while 19% used overly enthusiastic terms like "wonder pill". 39% of the 492 stories pointed to potentially harmful effects of the medication while 27% mentioned costs. 174 stories cited at least one expert describing the effects of the medication, but only four of these stories disclosed financial ties between the expert and the manufacturer of the specific drug. Press releases from drug companies were cited twice as often as papers in medical journals. INTERPRETATION: The Norwegian news media usually given new medications an overly enthusiastic coverage, while there is incomplete information about the benefits, risks and costs of the drugs as well as about the financial ties between medical experts and the pharmaceutical industry. PMID- 12555611 TI - [Care for refugees in sanctuary--a challenge for the health services]. AB - BACKGROUND: When the first sanctuaries for asylum seekers were established in 1993, psychological consequences were a concern and an interview study was started. MATERIAL AND METHODS: The study focused on the refugees' background, physical and psychological reactions, the children's situation, coping strategies and post-traumatic effects, and whether the church-asylants got adequate care from the health services. Twenty former and present church asylants and the same number of helpers with various backgrounds were interviewed. RESULTS: The reasons for seeking sanctuary in the church were fear for one's life, lack of future opportunities, and refusal of asylum. Uncertainty was the most difficult aspect of their plight. All 20 asylants reported headaches, insomnia, sadness, suicidal thoughts and restlessness. In a small minority suffering from severe psychiatric disturbances, previous experience of trauma, were predominant. Education, training, work, social contact and health assistance were organized to cope with the situation. Basic hopes, a wish to help children and family, and experience of friendship and solidarity were the most important motivational factors for keeping up morale. INTERPRETATION: The study shows that church asylants who take an active role, receive emotional support, and have good health services available are protected psychologically during their stay in the sanctuary, possibly also in the long term. PMID- 12555612 TI - [The physicians--homo economicus or homo sociologicus?]. AB - BACKGROUND: Different ways of organising and funding health services are based on certain assumptions about social action. Choice of theories affects the definition of the situation, policy content, goal fulfilment, as well as unintended effects. Here we discuss the role of the physician. How is it to be understood, and what implications do different theories and their accompanying political initiatives have for people's trust in doctors? MATERIAL AND METHODS: We start out from a theoretical discussion based on the economic and sociological theory of action in combination with three different perspectives: those of patients, of government/politicians, and those of the doctors themselves. RESULTS: It seems that patients see physicians as homo sociologicus; politicians see them as homo economicus; the physicians themselves are ambivalent towards their own role. INTERPRETATION: We should be careful not to design managerial systems that put the interests of patients and physicians in direct conflict, most of all because it could undermine the trust in the medical profession. PMID- 12555613 TI - [Axel Holst and Theodor Frolich--pioneers in the combat of scurvy]. AB - Axel Holst (1860-1931), professor of hygiene and bacteriology at the University of Oslo and paediatrician Theodor Frolich (1870-1947) became interested in a disease termed "ship beriberi" which afflicted the crews of sailing ships, and which showed an uncanny likeness to scurvy. They suspected a nutritional deficiency, and established an animal model that allowed systematic study of factors that led to disease as well as the preventive value of different substances. The choice of the guinea pig as the experimental animal for these studies was one indeed fortuitous, as that species has been shown to be among the very few mammals incapable of endogenous synthesis of ascorbic acid. They found that the guinea pigs developed distinctly scurvy-like symptoms when fed a diet consisting of various types of grain either whole or baked into bread, and that these symptoms were prevented when the diet was supplemented with known antiscorbutics like fresh cabbage or lemon juice. Their findings were published in 1907 in the Journal of Hygiene, but caused scientific uproar since the concept of nutritional deficiencies was a novelty at the time. The crucial factor, Vitamin C, was discovered in 1930 by Albert Szent-Gyorgyi, for which he was rewarded the Nobel Prize. No prizes or proper recognition were awarded Holst and Frolich at the time. It took some 60 years before they due acclaim was given to them; the 1907 paper by Holst and Frolich is now considered the most important single contribution to elucidating the aetiology of scurvy. PMID- 12555614 TI - [Obstetrical analgesia]. PMID- 12555615 TI - [Travelling to high altitude areas--acute high altitude sickness]. AB - BACKGROUND: People show increasing interest in travelling to high altitude areas such as the Himalayas, the Andes and mountains like Kilimanjaro in Tanzania. MATERIAL AND METHODS: Based on personal experience and published articles, an overview of acute high altitude sickness (acute mountain sickness) is given. RESULTS AND INTERPRETATION: High altitude sickness may be mild, moderate or severe, i.e. life threatening. There is a gradual shift in symptoms between these three syndromes. Mild to moderate high altitude sickness are recognized by various degree of headache, apathy, reduced appetite, nausea and vomiting, and when moderate, also peripheral oedemas and fluid retention. High altitude pulmonary oedema and high altitude cerebral oedema are life threatening conditions. High altitude sickness can be prevented by slow ascent. It is important to discover symptoms early. Descent is always the best treatment, but oxygen, portable pressure chambers and medication can be used in special situations. Acetazolamid is effective both to prevent and treat acute altitude sickness. Dexamethasone is an alternative, especially indicated for the treatment of high altitude cerebral oedema. Nifedipine is indicated in the treatment of high altitude pulmonary oedema. PMID- 12555616 TI - [Dangerous animals]. AB - As travellers seek ever more exotic destinations they are more likely to encounter dangerous animals. Compared to risks such as AIDS, traffic accidents and malaria, the risk is not so great; many travellers are, however, concerned about this and those who give pre-travel vaccines and advice should know something about it. This article is mainly based on medical and zoological textbooks. Venomous stings and bites may be prevented by adequate clothing and by keeping safe distance to the animals. Listening to those who live in the area is of course important. Travellers should not carry antisera with them, but antisera should be available at local hospitals. It should be borne in mind that plant eaters cause just as many deaths as large predators. In some cases it is necessary to carry a sufficiently powerful firearm. PMID- 12555617 TI - [Business trips and health]. PMID- 12555618 TI - [Updated when it comes to the prevention of restenosis?]. PMID- 12555619 TI - [Specialist services in connection with functional diseases]. PMID- 12555620 TI - [Peripartum idiopathic myocardiopathy: prospective echocardiographic study]. AB - Still frequent in Africa, the idiopathic peripartum cardiomyopathy (PPC) is the subject of only few prospective studies. The aim of this prospective work was to assess the echocardiographic abnormalities of the PPC and to determine on 26 patients the evolution and the prognostic factors of this disease. Six women had mild to moderate pericardial effusion. The abnormalities of the wall motion, constantly found, were diffuse in 20 cases (77.1%) and localized or prevalent on the interventricular septum or the left ventricular posterior wall in the other cases. The cardiac chambers were dilated in 24 cases (92.3%). Left ventricular hypertrophy was noted among 16 patients (61.5%); it was eccentric in 15 cases. The left ventricular systolic dysfunction was constant. The other abnormalities were: the abnormal left ventricular relaxation (one case), low mitral and aortic flow (12 cases), the mitral (21 cases) and tricuspid regurgitation (five cases). One noted a septal hypertrophy and an isolated dilatation of the right ventricle. The mean follow-up was 7.3 +/- 1 month (1-18 months). Two patients died at the 4th and 8th month. Among the 24 survivors, 11 had normalized were: the gestity (p = 0.01), the parity (p = 0.01), the cardiothoracic ratio (p = 0.04), the left ventricular volumes (p = 0.02), and the parameters of left ventricular function. The echocardiography of patients with PPC usually shows a pattern of dilated and hypokinetic cardiomyopathy, but many variations are possible. Most of the patients keep echocardiographic abnormalities after mean term follow-up. PMID- 12555621 TI - [Chronic constrictive pericarditis in African children: report of a case]. AB - The chronic constrictive pericarditis is a rare affection, with multiple etiologies and concerning especially the adult. We report a case of chronic constrictive pericarditis in an African child in whom no etiology was found. A review of the literature raises the characteristics of chronic constrictive pericarditis for a better therapeutic management. PMID- 12555622 TI - [Cardiac incidentaloma]. AB - We describe a case of lipomatous hypertrophy of the atrial septum mimicking a tumor of the right atrium. The association of echocardiography and radiology for the detection of this disease is illustrated in this report. PMID- 12555623 TI - [Indications of automatic ventricular implantable defibrillator. Implications for daily practice]. AB - The authors were the redactors of the Guidelines of the French Society of Cardiology for the indications of the automatic implantable defibillator, derived from the available indication in USA, and from the ulteriorly performed controlled studies. Three Class-I indications were selected: 1) circulatory arrest due to ventricular tachycardia (VT) or fibrillation (VF) whithout acute curable aetiology. 2) sustained VT with underlying heart disease and contractile alterations. 3) non-sustained VT with prior myocardial infarction and LVEF < 35% with inductible despite maximal drug therapy. Class-II indications were also three: 1) inheritable disease with high risk of sudden death without known effective therapy. 2) Syncope in patients with underlying heart disease and inductible VT or VF during electrophysiologic study. 3) VT or VF in patients in list for heart transplant. PMID- 12555624 TI - [Indications of atrial fibrillation regularization in 2001]. AB - The regularisation of the first episode of atrial fibrillation should be attempted. In the case of the second episode, the indication of the regularisation should be discussed according to the data of age, tolerance, objective haemodynamic effect, adequation to the treatment and possibilities of antiarrhythmic treatment. In fact, the prognosis of a correctly treated patient (with anticoagulants and slowing drugs) remaining in atrial fibrillation, does not seem to differ from those of a patient in sinus rhythm receiving antiarrhythmic drugs. PMID- 12555625 TI - [Analysis of the medico-economic literature comparing primary angioplasty and thrombolysis in the management of acute myocardial infarction]. AB - OBJECTIVE: To assess the generalizability of the medico-economic analysis comparing primary coronary angioplasty and thrombolytic therapy for acute myocardial infarction. METHOD: A systematic analysis of published studies was performed by two independent reviewers, in accordance with guidelines promulgated by health economic experts. RESULTS: Eleven articles, which concerned seven studies, were selected. Respectively, four evaluations were carried out in U.S. and three other in European countries (France, Netherland and Austria). There were three randomized trials, two observational studies and two decision trees. The costs were respectively ranged 2042 to 83,708 1999 US dollars for thrombolytic therapy and 3289 to 83,477 1999 US dollars for angioplasty. In two randomized trials and one decisional tree, the primary coronary angioplasty was both more effective and less costly than the thrombolysis therapy. One observational study concluded that thrombolytic therapy was less costly than primary angioplasty despite comparable effectiveness. Two analysis could not conclude of a difference between the alternatives, because of lack of statistical power. DISCUSSION: Published medico-economic analysis remain of a little interest for the French health care system because of lack of transparency in presentation of results. The dominance of the primary angioplasty was sensitive to time required for patient's transfer (ideally less than an hour), to the presence of redundant laboratories in an area and to the presence of an experienced staff for 24 h a day. PMID- 12555626 TI - [Quality of life and angina pectoris. Comparison of the effects of 2 nitrate derivatives administered discontinuously: transdermal 10 mg nitroglycerine patch (12 hours) and long-acting oral 40 mg isosorbide-5-mononitrate (once a day)]. AB - A comparative, randomised, 12-week (two periods of six weeks) cross-over study including 150 patients (mean age: 63.4 years) suffering from stable but symptomatic (a minimum of three attacks per week) angina pectoris was performed in order to compare the effect on quality of life of two discontinuous nitrate treatments: transdermal 10 mg nitroglycerine patch (12 hours) and long-acting oral 40 mg isosorbide-5-mononitrate (once a day). The efficacy and safety were also compared. The two treatments equally and significantly improved patients' quality of life. The number of attacks and sublingual nitrate consumption significantly decreased under treatment. Attack severity was lower under nitroglycerine than isosorbide-5-mononitrate treatment. Finally, even though nitroglycerine more frequently induced headache than did isosorbide-5-mononitrate (13 cases versus 8), patients, and in particular those having received nitroglycerine treatment in the second period of the study, preferred the transdermal nitroglycerine treatment. In conclusion, results did not show any relevant difference in terms of efficacy or tolerability between the two treatments. Nevertheless, the facility of use and the feeling of protection were better under transdermal nitroglycerine patch than oral isosorbide-5-mononitrate treatment. PMID- 12555627 TI - [Instable angina. Introduction]. PMID- 12555628 TI - [Physiopathology of unstable angina]. AB - The major risk of atherosclerotic disease is the occurrence of an acute coronary syndrome. The pathogenesis of instable angina involves the formation of an arterial thrombus as a consequence of the rupture of an atheromatous plaque. This risk of plaque rupture appears to depend on plaque morphology rather than plaque size or severity of stenosis. Ratio of lipid core to fibrous determined by the balance between smooth muscle cells proliferation and extracellular matrix synthesis stabilizing the plaque and macrophages which degrade collagen, determine the plaque vulnerability. The fibrous cap weakness leads to the plaque activation, plaque fissure or erosion activating a thrombotic cascade. A general inflammation or prothrombotic states are probably involved suggesting the need for a systemic therapeutic in addition with the treatment of the culprit lesion. PMID- 12555629 TI - [Antithrombotic therapy in the acute phase of unstable angina]. AB - Low molecular weight heparins appear to be a better choice in patients with unstable angina than unfractionated heparin. In addition, the excellent predictable dose-response makes them suitable to prevent ischemic complications of percutaneous coronary intervention. Although direct comparisons between LMWH and UH are limited, substantial evidence exists that patients receiving LMWH can be safely brought to cardiac catheterization. Therefore, previous concern regarding transitioning such therapy from the medical service to the cardiac catheterization laboratory should not impede the upstream use of these agents. Although UH remains an option in conjunction with GP IIB/IIIa inhibitors, there is substantial evidence that LMWH and GP IIb/IIIa inhibitor therapy can be used safely in combination. Although GP IIb/IIIa inhibitors are very potent to prevent ischemic events after percutaneous coronary interventions, their benefit in the medical management of unstable angina is much more modest. ADP receptor antagonists reduce major ischemic events in combination with aspirin in the medical management of unstable angina patients but also when PCI is planned. Their combination with GP IIB/IIIa receptor antagonist does not impair the benefit of GP IIb/IIIa antagonists. The development of biologic tools to monitor these antithrombotic regimen is highly warranted especially in high risk patients (chronic renal failure, elderly). PMID- 12555630 TI - [Glycoprotein IIb/IIIa inhibitors in unstable angina]. AB - During the last ten years we have considerably update our knowledge about the pathogenesis of unstable angina or acute coronary syndromes. The platelet activity have in this settings the central role in development of the thrombotic process. Platelet glycoprotein IIb/IIIa inhibitors block fibrinogen binding to platelets, and the effect of this on the final common pathway of platelet aggregation makes these compounds extremely potent antiplatelet drugs. Three intravenous IIb/IIIa receptor antagonists are approved for clinical use, and this class of therapy has update our pharmacologic armatarium to avoid ischemic complication in the settings of percutaneous coronary revascularization at first and now in medical treatment of acute coronary syndromes. Results of large trials using this drugs suggest that this agents are effective in patient with unstable angina particularly in those presenting a high score of risk for acute ischemic events and those requiring coronary intervention. PMID- 12555631 TI - [Treatment of unstable angina: early invasive strategy]. AB - Unstable angina is one of the main reasons for admission to a cardiology department and the risk of myocardial infarction is high. Non-Q wave myocardial infarction used to be assimilated to unstable angina. Myocardial infarction without prolonged ST segment elevation and with troponin increase is now the category used to describe this condition. CPK elevation, EKG and clinical parameters are involved in the acute prognosis of patients with unstable coronary syndrome. The FRISC II trial was the first study to demonstrate the superiority of systematic early angiography followed by a revascularization procedure selected according to the type of lesions to be treated, compared with conventional management. The use of stents combined with GPIIb/IIIa inhibitors has increased the safety of PTCA and seems to have a synergetic effect. The superiority of early invasive management using stents and GPIIb/IIIa inhibitors was confirmed by the TACTICS. The risks of bleeding complications associated with this therapeutic strategy are minimized by the TIMI classification. This classification no longer corresponds to the current norms of patient safety and comfort. Several methods can be used in order to decrease the risk of bleeding complications; the radial approach is undoubtedly the most efficient one. PMID- 12555632 TI - [Unstable angina in the elderly]. AB - Unstable angina and acute coronary syndromes without persistent ST-segment elevation are frequent and their prognosis is poor in the elderly. Indeed, age is the most powerful predictor of in-hospital mortality in this setting. The clinical benefit of interventional strategies, as demonstrated by FRISC II and TACTICS TIMI 18 studies, seems to be most important in this age subset. PURSUIT trial demonstrates that the efficacy of eptifibatide, a IIb/IIIa platelet receptor inhibitor, increases in elderly patients who simultaneously undergo revascularization interventions. Individual application of such treatment strategies may be difficult. Potential triggering factors of unstable angina and comorbidities have to be taken into account, and the overall management should be highly individualized in elderly patients. The aim remains to achieve appropriate myocardial revascularization, as often as possible by focusing coronary angioplasty on the culprit vessel. Coronary surgery generally should be reserved for coronary lesions which are not suitable for percutaneous revascularization. Clinical improvement is maximal in patients with severe initial presentation. PMID- 12555633 TI - [Prognostic value of normal coronary angiography]. AB - Coronary angiography is the "gold standard" for coronary artery disease (CAD). It is considered either normal or subnormal without any lesion (endocoronary echography often demonstrates atheroma), or in presence of a < 50% stenosis. Nevertheless, the risk of plaque rupture is not well correlated with the degree stenosis. Despite the frequent presence of non-significant atheroma, is a normal coronarography really of a good prognosis? Between January and September 1997, 136 of 600 (22.6%) angiographies were considered as normal. The indications were: "CAD suspicion" (n = 77), "preoperative angiography of valvulopathy" (n = 38), and "angioplasty control" (n = 22). The arteries were strictly normal for 86 patients (63%) and a < 50% stenosis was found in 50 patients (37%); 108 patients (80.1%) were followed for 18 +/- 3 months: eight non coronary deaths were reported: four postoperative deaths in "valvular group", two pulmonary embolisms and two pulmonary neoplasm's in "CAD suspicion group". No myocardial infarction was reported and one unstable angina was documented. Despite the frequency of non significant atheroma, an acute coronary syndrome exceptionally complicates a "normal" coronarography. PMID- 12555634 TI - [Mobile and obstructive thrombus of the left atrium. Value and risks of transesophageal echocardiography. Report of a case and review of the literature]. AB - Authors report the observation of a young woman of 30 years hospitalised for degeneration of a mitral biologic prosthesis. Transthoracic echocardiography visualizes a voluminous mass of the left auricle of which the character free and destitute of all parietal attachment, doesn't have can be established that by transoesophageal echocardiography that detects another thrombus also free and mobile of the left auricule, unrecognised by transthoracic way. The operative indication is carried in emergency but the patient dies suddenly some hours after this exam. From this observation and a revue of the literature, the contribution and risks possible transoesophageal echocardiography in the exploration of the mobile thrombi of the left auricle, are analysed. Account-held of the potential gravity of homodynamic and embolic complications, the emergency of the surgical indication is underlined. PMID- 12555635 TI - [Heart involvement in carcinoid syndrome: report of a case]. AB - The authors report the case of tricuspid and pulmonary disease, revealing a carcinoid syndrome in a 32 years-old young man who was admitted for asthenia and an effort hepatology. The carcinoid syndrome was confirmed by hormonal proportioning, and by an anatomopathologic and immunohistochimic study of a hepatic metastatic biopsy. The check-up to search the primitive tumor was negative. The clinicals, paraclinicals and therapeutics aspects of the carcinoid heart were reported in this work. PMID- 12555636 TI - [Rehabilitation modalities as a function of the severity of heart insufficiency]. AB - After a review of the different central and peripherical factors limiting exercise capacity in chronic heart failure, the authors report the mechanisms of these increment with exercise training. The different indications for clinical training techniques of segmental training on a specific bench are discussed; but it appears that both aerobic and repetitive weight programs seem to have short term favorable and complementary effects. PMID- 12555637 TI - [Heart insufficiency in the elderly]. AB - Congestive heart failure in the elderly differs from the one in the younger. The ageing of the cardiovascular system makes the organism weaker. When a myocardial infarction or an other cardiovascular disease happens, the occurrence of congestive heart failure is precipitated. The symptoms which are often misleading and the polypathologies make the assumption of relationship difficult between a symptom like dyspnea and congestive heart failure. Further examinations are limited because of the reduced physical performances in the elderly (stress test) or because of an increased risk of side-effects (coronary angiogram). The echocardiography has a central role in the exploration of congestive heart failure. The medical treatment has the same principles than in the younger but with cautions especially regarding the renal insufficiency and the multiple treatments that an elderly patient has. PMID- 12555638 TI - [Norwegian prioritization]. PMID- 12555639 TI - [Thrombolytic therapy in stroke--just for some?]. PMID- 12555640 TI - [When death is better than life]. PMID- 12555641 TI - [Centralized treatment of rare types of cancer]. PMID- 12555642 TI - [Bone and soft tissue sarcomas treated at the Norwegian Radium Hospital 1980-99]. AB - BACKGROUND: The Norwegian Radium Hospital's sarcoma group is a multidisciplinary group with a leading role in the diagnosis and treatment of bone and soft tissue sarcomas in Norway. MATERIAL AND METHODS: From 1980 through 1999, 1,355 patients with soft tissue sarcoma and 458 patients with bone sarcoma were treated. In a retrospective analysis of trends over time, patients were allocated to consecutive five-year periods. RESULTS: Patient characteristics were relatively stable, but there was an increasing proportion of soft tissue sarcomas being referred without prior surgery. Treatment principles have remained unchanged, with surgery with or without radiotherapy dominating in soft tissue sarcoma and surgery with or without chemotherapy in bone sarcoma. The amputation rate for bone sarcoma has fallen from 78% to 17%, and survival has increased significantly for both soft tissue and bone sarcoma patients. INTERPRETATION: The results indicate significant improvements in the quality of treatment of soft tissue and bone sarcoma. More resources for treatment and organizational development of a multidisciplinary group may contribute to improved quality of care. PMID- 12555643 TI - [Thrombolytic therapy in acute cerebral infarction]. AB - BACKGROUND: The aim of this study was to assess the feasibility, safety, and clinical results in patients treated with intravenous thrombolysis in an acute stroke unit. Results from the first three years with thrombolysis as standard treatment procedure are presented. MATERIAL AND METHODS: All patients admitted to our department within 3 hours after an acute ischaemic stroke were considered for thrombolysis. 33 patients with median age 71 years were treated. RESULTS: 16 patients demonstrated early clinical improvement. Six patients demonstrated a substantial but slow clinical improvement. 11 patients did not improve. One patient developed an intracerebral haematoma and two developed a haemorrhagic infarction without clinical deterioration. At follow-up after six months, 16 patients (49%) had achieved independence [modified Rankin Scale (mRS) 0-2], 12 (36%) had an unfavourable outcome (mRS 3-5). Five patients (15%) died, all within the first 3 months. None died due to a treatment complication. INTERPRETATION: This study suggests that thrombolysis can be administered relatively safely in an acute stroke unit without intensive care facilities. The clinical results and safety were similar to those in large randomized studies and clinical series. PMID- 12555644 TI - [Parents' experiences with life-and-death decisions concerning their premature infants]. AB - BACKGROUND: The study is concerned with parents' experience with life-and-death decisions concerning their premature infants in hospitals neonatal units. What are parents' attitudes towards their being involved in such decision-making? MATERIAL AND METHODS: During the 1997-2000 period, 20 qualitative interviews with 35 parents of 26 children were made. A qualitative, comparative method (grounded theory) was used for the analysis, which was carried out in parallel with data collection. RESULTS: Parents are exposed to great mental pressure, which may bring about a sense of ambivalence and a wish to avoid responsibility. Most parents do not wish to have the final word in decisions concerning their infant's life or death. Such a responsibility is likely to make parents feel guilty and puts too heavy a burden on the parents, but parents wish to be informed and taken seriously in the decision making-process. INTERPRETATION: Parents value health professionals' communication and information more than parent autonomy and right to choose. PMID- 12555645 TI - [Preoperative expectations and clinical outcome of transmyocardial laser treatment in patients with angina pectoris]. AB - BACKGROUND: The aims of this study were to evaluate the effect of transmyocardial laser treatment on quality of life and to assess the correlation between preoperative expectations and clinical improvement after one year. MATERIAL AND METHODS: 13 patients (median age 56 years) with disabling angina pectoris were subjected to transmyocardial holmium: YAG laser. Quality of life was assessed preoperatively and at three and 12 months by Hospital Anxiety and Depression Scale (HAD), Physical Symptom Distress Index (PSDI) and Life Satisfaction Index (LSI). Expectations were evaluated by Leedham's scale. RESULTS: A significant improvement in Canadian Cardiovascular Society Score (CCS) from 3.4 +/- 0.5 (mean +/- SD) preoperatively to 1.6 +/- 1.0 and 1.7 +/- 0.8 three and 12 months after treatment was observed (p < 0.01). Quality of life (PSDI and LSI) improved. No significant changes in ejection fraction or exercise performance were found. Preoperative expectations were generally high, but did not correlate significantly with improvements in CCS or quality of life. INTERPRETATION: Although no changes in objective parameters were found, the lack of significant correlations between preoperative expectation and subjective clinical improvement indicate that the improvement of angina pectoris only partly can be explained by placebo effects. PMID- 12555646 TI - [Screening for dysplasia of the hip at health centers]. AB - BACKGROUND: Screening for developmental dysplasia of the hip has a long tradition in health centres in Norway. Under the guidelines from Norwegian health authorities, the presence of asymmetric abduction of the hips should be the most important indication for referral to ultrasound examination. The outcome of this procedure is not well known, however. MATERIAL AND METHODS: Throughout the year 2000, all children referred for ultrasound examination of their hips to Ullevaal University Hospital and Oppland Central Hospital Lillehammer were recorded prospectively. The indication for and outcome of the examination of children referred from health centres were examined. RESULTS: 133 girls and 92 boys were referred. Three girls and three boys were identified with dysplasia of the hip, giving an estimated prevalence of 0.5 pr 1,000. Asymmetric abduction of the hip was the most frequent reason for referral with an estimated positive predictive value of 3%. 50 children were examined due to asymmetric femoral skin folds that were probably the only reason for suspecting dysplasia. Two of these children had dysplasia of the hip. 16 children had immature hips necessitating renewed examination, often identifying normal hips. INTERPRETATION: The screening for developmental dysplasia of the hip at health centres is based on clinical tests or criteria with low positive predictive values. PMID- 12555647 TI - [Bone adaptation to mechanical loading]. AB - BACKGROUND: In general, regular physical activity has a strong effect in promoting health and preventing diseases. Bone health is promoted through regular weight-bearing physical activity that uses muscular strength and power and exerts force on the skeleton above normal levels. Some theories have been advanced to explain the relationship between mechanical loading and the bone tissue's biological response. The aim of this review is to review some of these theories and discuss how bone adapts to mechanical loading. MATERIAL AND METHODS: The literature was identified on Medline. In addition, relevant books were examined. RESULTS: The mechanostat theory is widely accepted. It is based on the assumption that bone structure changes through a feedback system in which changes in peak mechanical strain drive bone cells to change bone structure. Other theories suggest that bone formation depends upon the number of strain cycles, strain distribution and strain rate, not only the strain magnitude as suggested in the mechanostat theory. INTERPRETATION: The magnitude of the loading, type of activity, the rate of the activity, and the number of repetitions seem to be important factors associated with the effect of physical activity on bone. PMID- 12555648 TI - [Does exercise improve the skeleton of young women?]. AB - BACKGROUND: High bone mineral density (BMD) at menopause can reduce the risk for osteoporotic fractures later in life, hence it is important to identify factors that can increase or maintain BMD during the premenopausal years. Physical activity is considered a major factor. This review examines how physical activity affects BMD in premenopausal women. MATERIAL AND METHODS: The literature was identified on Medline. In addition, relevant books were examined. RESULTS: Research indicates that weight-bearing exercises that involve high-magnitude loading can maintain or increase BMD in premenopausal women, especially in the loaded bones. Low to moderate intensive training and non-weight bearing exercise show minor or no effect on BMD. INTERPRETATION: In every day life, effective physical activity or training to maintain or increase bone mass in young women ought to include short-term high mechanical load, preferably with unusual load distribution, which also is fast and dynamic. Physical activity in young women seems to be an important factor in reducing the risk of osteoporotic fractures later in life. PMID- 12555649 TI - [Acetylsalicylic acid against pre-eclampsia?]. PMID- 12555650 TI - [Schizophrenia and molecular genetics]. AB - BACKGROUND: Schizophrenia is a severe mental disorder with a lifetime risk in the population of 1%. Despite extensive research, the underlying neurobiological mechanisms remain largely unknown. MATERIAL AND METHODS: Based on relevant research literature, an overview of the molecular genetics of schizophrenia is given. RESULTS AND INTERPRETATION: Family, twin and adoption studies have shown that schizophrenia has a high heritability, indicating a significant genetic component to its aetiology. There is positive linkage to chromosomal regions which probably contain susceptibility genes of moderate effect. Genetic variants in different candidate genes and different chromosome abnormalities are apparently associated with increased susceptibility to schizophrenia. However, the genetic mechanisms of schizophrenia are still largely unknown. As a result of recent advancements in molecular genetic knowledge and methods, high-throughput techniques are now available in functional genomic research that will enable genome-wide search for susceptibility genes. This has to be combined with a new clinical approach where schizophrenia is characterized based on symptoms and traits that might more directly reflect fundamental processes of relevance to the disorder. PMID- 12555651 TI - [Neurobiological findings in schizophrenia]. AB - BACKGROUND: Schizophrenia is a syndrome with important neurobiological abnormalities which have not yet been fully clarified and whose aetiology is poorly understood. MATERIAL AND METHODS: The paper gives a survey of the most important neurobiological abnormalities found in schizophrenia, based on relevant research publications, mainly from the most recent decades. RESULTS AND INTERPRETATIONS: Pathomorphological findings in schizophrenia include ventricular and cortical sulcal enlargement, reduction of cortical thickness, cellular changes with reduction of dendrites and synapses, presumably resulting in reduced cortico-cortical and cortico-subcortical connections. These abnormalities are thought to be mainly developmental in origin, mostly on a genetic basis, but also with varying contributions from prenatal or perinatal damage. In a subgroup of schizophrenics there is a poorly understood neurodegenerative process going on also after the debut of the psychosis. Neurochemically, interest has focused mostly on dopaminergic dysfunction, but other transmitters like serotonin, glutamate and GABA are also involved, as is probably the neurotensin peptide. There is also increasing evidence of disturbed cell membrane composition and function in schizophrenia, especially with regard to polyunsaturated fatty acids. PMID- 12555652 TI - [Medicine, law and tobacco industry's freedom from responsibility]. PMID- 12555653 TI - [Diagnosis of high risk of heart disease among asymptomatic persons]. PMID- 12555654 TI - [How common is familial hypercholesterolemia?]. PMID- 12555655 TI - [Misleading information about statins]. PMID- 12555656 TI - [English in teaching]. PMID- 12555657 TI - [Call for more health]. PMID- 12555658 TI - Reversal of the adult IgE high responder phenotype in mice by maternally transferred allergen-specific monoclonal IgG antibodies during a sensitive period in early ontogeny. AB - IgE is an important trigger in allergy and asthma, diseases whose development is suggested to depend on an initial sensitization in early life. While induction of murine IgE responses requires both a genetically based IgE high responder phenotype and defined experimental conditions, maternally transferred IgG can override these prerequisites and suppress IgE formation in an allergen-specific manner. Here, we show that maternally transferred monoclonal IgG, irrespective of their subclass and recognized epitopes, induce IgE unresponsiveness, which is effective for parenteral immunization with bee venom phospholipase A2 as well as for airway-immunization with nebulized ovomucoid-containing ovalbumin. This IgE suppression is detectable in the offspring during the first 4 months of life, but not thereafter and not in the dams. However, when the initial immunization at an age of 3 or 4 months was followed by further application of both allergens via their respective routes, IgE suppression persisted up to an age of more than one year. If applicable to man, these findings may allow the development of a new strategy for the prevention of allergy and asthma by maternally transferred or neonatally injected allergen-specific mAb in combination with natural or prophylactic exposure to the respective allergens during early childhood. PMID- 12555659 TI - Tumor necrosis factor-receptor 2 is up-regulated on lamina propria T cells in Crohn's disease and promotes experimental colitis in vivo. AB - Tumor necrosis factor (TNF) plays a pivotal role in the pathogenesis of Crohn's disease (CD). However, little is known about the role of TNF receptors (TNF-R) in this disease. Here, we found that TNF-R2 (in contrast to TNF-R1) was significantly up-regulated on lamina propria and peripheral blood T cells in CD compared to control patients. To directly test the functional role of TNF-R2 in Th1-mediated experimental colitis in vivo, we took advantage of transgenic animals overexpressing TNF-R2 in T cells. Reconstitution of SCID mice with CD4+ CD62L+ T cells from TNF-R2 transgenic mice led to an earlier wasting syndrome, a more severe colitis and augmented Th1 cytokine production than reconstitution with cells from wild-type littermates. In addition, TUNEL staining revealed a significantly decreased apoptosis rate of lamina propria mononuclear cells in mice reconstituted with TNF-R2 transgenic T cells compared to mice reconstituted with wild-type T cells. In summary, our data suggest a critical regulatory role of TNF-R2 signaling for disease exacerbation in Th1-mediated chronic colitis. Taken together with the increased expression of TNF-R2 in CD, selective targeting of TNF-R2 signaling thus emerges as a potentially novel approach to the treatment of CD. PMID- 12555660 TI - DNA polymerase kappa deficiency does not affect somatic hypermutation in mice. AB - Somatic hypermutation (SH) in B cells undergoing T cell-dependent immune responses generates high-affinity antibodies that provide protective immunity. Most current models of SH postulate the introduction of a nick into the DNA and subsequent replication-independent, error-prone short-patch synthesis by one or more DNA polymerases. The Pol kappa (DinB1) gene encodes a specialized mammalian DNA polymerase called DNA polymerase kappa (pol kappa), a member of the recently discovered Y family of DNA polymerases. The mouse PolK gene is expressed at high levels in the seminiferous tubules of the testis and in the adrenal cortex, and at lower levels in most other cells of the body including B lymphocytes. In vitro studies showed that pol kappa can act as an error-prone polymerase, although they failed to ascribe a clear function to this enzyme. The ability of pol kappa to generate mutations when extending primers on undamaged DNA templates identifies this enzyme as a potential candidate for the introduction of nucleotide changes in the immunoglobulin (Ig) genes during the process of SH. Here we show that pol kappa-deficient mice are viable, fertile and able to mount a normal immune response to the antigen (4-hydroxy-3-nitrophenyl)acetyl-chicken gamma-globulin (NP-GC). They also mutate their Ig genes normally. However, pol kappa-deficient embryonic fibroblasts are abnormally sensitive to killing following exposure to ultraviolet (UV) radiation, suggesting a role of pol kappa in translesion DNA synthesis. PMID- 12555661 TI - Self peptide/MHC class I complexes have a negligible effect on the response of some CD8+ T cells to foreign antigen. AB - MHC molecules loaded with self peptides do not trigger a T cell immune response but may deliver signals important for peripheral T cell survival and function. It is unclear if self peptide/MHC complexes on APC in addition can influence the T cell response to co-presented foreign ligands. To address this question, TAP sufficient and TAP-deficient cells were loaded with ovalbumin peptide (pOVA) to generate APC that present pOVA/H-2Kb complexes in the context of high or low levels of self peptide-loaded MHC class I, respectively. The two cell types were then used to stimulate different CD8+ T cells specific for ovalbumin while the number of presented pOVA/H-2Kb complexes was independently assessed by staining with 25-D1, an antibody against pOVA/H-2Kb. In each case, T cell activation was independent of TAP expression by the APC and depended exclusively on the amount of 25-D1 staining. We conclude that the number of pOVA/Kb complexes and not their frequency relative to self peptide/MHC complexes determines the response of those T cells tested here. These results imply that the repertoire of self peptide/MHC class I complexes presented by APC has a negligible effect on the response of some CD8+ T cells to foreign ligands. PMID- 12555662 TI - Targeting CLA/E-selectin interactions prevents CCR4-mediated recruitment of human Th2 memory cells to human skin in vivo. AB - Naive Th cells, bearing receptors for cutaneous antigens, become activated in skin-draining lymph nodes and express cutaneous lymphocyte antigen (CLA), which confers to these cells the capacity to migrate into the skin to exert their normal effector functions. In the case of atopic dermatitis (AD), allergen specific Th2 cells generate exacerbated responses and induce skin inflammation. In such a situation, interfering with the specific mechanism of skin homing would provide a therapeutic benefit. Here we report that CLA+ Th2 memory cells, derived from skin lesions of AD patients, selectively migrate to human skin grafts transplanted onto SCID mice in response to CCR4 but not CCR3, CCR8 or CXCR3 ligands. Skin homing of human CCR4+ Th2 memory cells was Pertussis toxin sensitive and restricted to the CLA+ subset. Furthermore, treatment of these mice with anti-E-selectin monoclonal antibody was sufficient to prevent CCL22-mediated Th2 cell migration to human skin, which both, validates the model and highlights the importance of CLA/E-selectin interactions in the homing process of Th2 cells to the skin. Using this mechanistic model we demonstrate that skin homing of human Th2 memory cells can be efficiently suppressed using a low molecular weight E-selectin antagonist, which is of clinical relevance for the treatment of inflammatory skin diseases, including AD. PMID- 12555663 TI - Dominant TCR V alpha usage by virus and tumor-reactive T cells with wide affinity ranges for their specific antigens. AB - We have studied the TCR features and functional responses of three sets of human cytolytic T cell (CTL) clones, recognizing antigenic peptides presented by HLA-A2 and derived from the Epstein-Barr virus proteins BMLF1 and BRLF1 and from the melanoma protein Melan-A/MART-1. Within each set, a majority of clones used a recurrent V alpha region, even though they expressed highly diverse TCR beta chains and V(D)J junctional sequences. Functional assays and peptide/MHC multimer binding studies indicated that this restricted V alpha usage was not associated with the affinity/avidity of the CTL clones. The V alpha dominance, which may be a frequent feature of antigen-specific T cells, likely reflects a restricted geometry of TCR/peptide/MHC complexes, primarily determined by V alpha CDR. PMID- 12555664 TI - Structural and functional differences between putative mucosal inductive sites of the rat. AB - Nasal-associated lymphoid tissue (NALT), bronchus-associated lymphoid tissue (BALT) and Peyer's patches (PP) were compared structurally and functionally using a model of local mucosal infection of rats with reovirus. Histological analyses showed that BALT lacks the typical lymphoid organization found in NALT and PP. After local reovirus infection, germinal centers developed in NALT with appearance of IgA+ cells, whereas no germinal centers or isotype-switched cells were found in BALT. Production of reovirus-specific IgA was observed in NALT and PP, but only small amounts of specific IgA were secreted by BALT. Both NALT and BALT showed considerable production of IgG2b, whereas this isotype was poorly produced by PP. These data reveal profound qualitative differences between these three mucosal sites, and strongly suggest that BALT is not a mucosal inductive site for reovirus-specific immune responses in the rat. PMID- 12555665 TI - Blockade of CXCL10 protects mice from acute colitis and enhances crypt cell survival. AB - Crypt cell renewal is essential for normal intestinal homeostasis as well as mucosal regeneration following injury. However, the factors regulating crypt cell growth in pathological conditions are not fully understood. We report here that the endogenously produced chemokine CXCL10 regulates crypt cell proliferation. CXCL10 was constitutively expressed by basal crypts in mouse colon, but the expression of CXCL10 as well as CXCR3 was enhanced in the epithelium in the proliferative zone after oral administration of dextran sulfate sodium. Neutralization of CXCL10 protected mice from epithelial ulceration by promoting crypt cell survival without evidence of altered immune cell infiltration. Furthermore, recombinant CXCL10 administration into mice inhibited intestinal epithelial cell proliferation. These findings suggest that CXCL10 regulates crypt cell growth to maintain intestinal homeostasis in an autocrine or paracrine fashion. Thus, CXCL10 can be a new therapeutic target for inflammatory bowel disease by controlling the dynamics of epithelial homeostasis. PMID- 12555666 TI - Control of New World cutaneous leishmaniasis is IL-12 independent but STAT4 dependent. AB - Leishmania mexicana, a New World protozoan parasite, induces small, chronic, but non-progressive lesions in C57BL/6 (B6) mice. In this study we investigated the role of IL-12, and subsequent Th1 factors, in controlling cutaneous L. mexicana infection. IL-12 treatment failed to promote disease resolution, suggesting that the inability of mice to heal is not related to a deficiency of endogenous IL-12 production. Surprisingly, L. mexicana-induced cutaneous lesions in wild-type and IL-12p40-deficient mice were indistinguishable, with similar parasite burdens, immune responses, and lesion histopathology. In contrast, iNOS, IFN-gamma, and STAT4-deficient mice developed progressive disease and uncontrolled parasite growth. These results differ dramatically from L. major infection, in which IL 12p40-deficient mice are highly susceptible, with very rapid lesion growth, very large parasite burdens, and the development of a strong Th2 response. These data uncover the existence of an alternate IFN-gamma and iNOS pathway for control of Leishmania lesions, which is IL-12 independent, but which unexpectedly requires STAT4. PMID- 12555667 TI - The IL-10-producing competence of Th2 cells generated in vitro is IL-4 dependent. AB - Previous studies have suggested that activation of CD4+ T cells in the presence of IL-10 results in the generation of a population of T cells, T regulatory 1 (Tr1) cells, that primarily produce IL-10 and TGF-beta, but not IL-4. The relationship between Tr1 cells and conventional Th2 cells remains unclear. We were not successful in our attempts to generate significant numbers of antigen specific T cells that secreted IL-10, but not IL-4, by culture in the presence of IL-10. The small numbers of cells that produced IL-10 only were completely dependent on the presence of IL-4 for their generation. In a polyclonal model, the development of IL-10 only producers was completely dependent on the presence of signal transducer and activator of transcription 6. Studies with myelin basic protein-specific T cells derived from an IL-4-deficient mouse confirmed the absolute requirement for IL-4 for the generation of IL-10 producers under all culture conditions. These IL-10-producing Th2 cells failed to inhibit EAE in an adoptive transfer model and were pathogenic when transferred to immunodeficient mice. Collectively, our results raise doubts about the existence of a unique population of CD4+ regulatory T cells that can be generated by activation in the presence of IL-10. PMID- 12555668 TI - Pre-B-cell colony-enhancing factor, whose expression is up-regulated in activated lymphocytes, is a nicotinamide phosphoribosyltransferase, a cytosolic enzyme involved in NAD biosynthesis. AB - The murine homologue of the previously identified human "pre-B-cell colony enhancing factor" (PBEF) gene coding for a putative cytokine has been identified by screening a subtractive library enriched in genes expressed in activated T lymphocytes. Unlike most cytokine genes known to date, the PBEF gene is ubiquitously expressed in lymphoid and non-lymphoid tissues and displays significant homology with genes from primitive metazoans (marine sponges) and prokaryotic organisms. Recently, a bacterial protein encoded by nadV, a gene from the prokaryote Haemophilus ducreyi displaying significant homology with PBEF, has been identified as a nicotinamide phosphoribosyltranferase (NAmPRTase), an enzyme involved in nicotinamide adenine dinucleotide (NAD) biosynthesis. Using a panel of antibodies to murine PBEF, we demonstrate in this work that, similarly to its microbial counterpart, the murine protein is a NAmPRTase, catalyzing the condensation of nicotinamide with 5-phosphoribosyl-1-pyrophosphate to yield nicotinamide mononucleotide, an intermediate in the biosynthesis of NAD. The role of PBEF as a NAmPRTase was further confirmed by showing that the mouse gene was able to confer the ability to grow in the absence of NAD to a NAmPRTase-defective bacterial strain. The present findings are in keeping with the ubiquitous nature of this protein, and indicate that NAD biosynthesis may play an important role in lymphocyte activation. PMID- 12555669 TI - Combined dendritic cell- and CpG oligonucleotide-based immune therapy cures large murine tumors that resist chemotherapy. AB - The use of dendritic cells (DC) loaded with tumor antigen is one of the most advanced approaches in cancer immunotherapy. CpG motifs within microbial DNA detected by toll-like receptor 9 are responsible for the favorable properties of CpG oligodeoxynucleotides (ODN) as immune modulators. In this study, mature antigen-pulsed DC or peritumoral injections of CpG ODN, both effective for the treatment of small established tumors, were almost ineffective against large established tumors (1-cm diameter) in a syngeneic murine colon carcinoma model. For large tumors, the antitumor activity of mature antigen-pulsed DC was strongly increased by coinjection of CpG ODN, resulting in a transient control of tumor growth. Rejection of large tumors and long-term cure of mice was achieved by combining injection of antigen-pulsed DC plus CpG ODN at a site distant to the tumor with peritumoral injections of CpG ODN. Depletion of CD8 T cells abrogated the therapeutic activity. Large numbers of DEC-205-positive DC infiltrated the tumor in treated mice. Therapy with 5-fluorouracil and leucovorin was unable to control tumors of the same size. In conclusion, we demonstrate that the immune system, provided that appropriate stimulation with DC and CpG ODN is given, has the potential to cure animals of large solid tumors in situations where even chemotherapy is not efficient. PMID- 12555670 TI - Rapid constitutive generation of a specific peptide-MHC class II complex from intact exogenous protein in immature murine dendritic cells. AB - MHC class II molecules present peptides, derived largely from exogenous antigens, to CD4+ T cells. Complex-generation occurs mainly in the specialized late endosomal MHC class II-rich compartment (MIIC) vesicles of antigen-presenting cells (APC). Dendritic cells (DC) have been reported to store intact antigen in MIIC until the receipt of an activation signal, when they process it into peptide MHC class II complexes. However, constitutive migration of DC from the periphery to secondary lymphoid organs has been observed, and antigen presentation by nonactivated DC is proposed to play a role in the induction of tolerance to peripheral antigens. Thus, constitutive peptide-MHC class II complex generation must also occur in DC in immunologically quiescent situations. We have used a monoclonal antibody that detects a specific peptide-MHC class II complex to directly demonstrate constitutive complex generation in immature murine DC. Protein-derived peptide-MHC class II complexes were detected by flow cytometry at the DC surface within 1 h of antigen exposure in the absence of an exogenous activation signal, and could be detected by confocal microscopy in the MIIC within 5 min of antigen exposure. This processing activity was endotoxin independent. These data provide evidence for constitutive peptide-MHC class II complex generation in immature DC, and thus support a role for this activity in the induction of peripheral tolerance. PMID- 12555672 TI - Depletion of CD25+ regulatory cells uncovers immune responses to shared murine tumor rejection antigens. AB - Although it is known that the immune system can mount responses to a variety of tumors it is clear that most tumors exhibit weak or even undetectable immunogenicity. Recent findings suggest that the lack of tumor immunogenicity is partly due to a population of cells called CD4+ CD25+ regulatory T cells since depletion of these cells in mice can result in tumor rejection. These cells have also been shown to inhibit the development of organ-specific autoimmune diseases suggesting that they inhibit immune responses to tissue-specific self-antigens. Such immune responses may also mediate tumor rejection. Alternatively, immune responses in mice depleted of regulatory cells may target tumor antigens that are not tissue-specific, but which are shared by tumors of diverse origins. In experiments performed to discriminate between these possibilities we found, using the murine colorectal tumor CT26, that tumor immunity stimulated in the absence of regulatory cells is not restricted to tumors of colorectal origin, but is effective against tumors of different histological types such as B cell lymphomas and a renal cell carcinoma. By comparing this to CT26-induced immunity through the use of adjuvant we show that the generation of cross-reactive tumor immunity is a specific manifestation of CD25+ regulatory cell depletion. The generation of CD4+ T cells capable of mediating tumor rejection is another important feature of tumor immunity induced in the absence of CD25+ cells. PMID- 12555671 TI - Disruption of the lineage restriction of TCR beta gene rearrangements. AB - Despite a common lymphoid recombinase, assembly of Ig genes is restricted to B cells, whereas TCR loci rearrange in T cells. Transcriptional promoters and enhancers are critical for the regulation of the recombination process. However, the specific function of such elements in conferring the lineage-restriction of V(D)J recombination remains poorly understood. To gain further insights into the mechanism restricting TCR beta-chain rearrangements to T cells, we generated mice in which an 11 kb region--containing the beta-chain constant region 2 and the TCR beta enhancer (E beta)--was replaced with the B cell specific Ig heavy-chain enhancer (E mu). Unlike the simple E mu to E beta replacement, this mutation allowed significant levels of D beta to J beta as well as V beta to DJ beta rearrangements in both T and B cells. Although the lineage restriction was disrupted, TCR beta allelic exclusion was still efficient in mutated T cells. Together these results demonstrate that changes in the activity of regulatory elements located at the TCR beta constant regions are sufficient to redirect the recombination pattern of TCR beta variable gene segments. PMID- 12555673 TI - RANTES (CCL5) production during primary respiratory syncytial virus infection exacerbates airway disease. AB - Respiratory syncytial virus (RSV) is a respiratory pathogen that causes significant morbidity in infants and young children. The importance of chemokines during RSV infection for respiratory symptoms has not been fully elucidated. The current study examined the effect of RANTES (CCL5) on airway pathophysiology after RSV infection. BALB/c mice produce RANTES (CCL5) after RSV infection that correlates with the changes in pathophysiology. Animals treated with anti-RANTES (CCL5) antibody demonstrated significant decreases in airway hyperreactivity (AHR). Delayed treatment with anti-RANTES (CCL5) at day 5 of infection also significantly reduced development of AHR on day 9 of infection, suggesting that RANTES (CCL5) may be a target in established disease. Determination of Th1/Th2 associated cytokine patterns indicated that anti-RANTES (CCL5) treatment increased IL-12 production, thus altering the lung environment. The assessment of RANTES (CCL5) production in vitro and in vivo demonstrated that it was regulated by IL-13, a cytokine that is related to RSV-induced AHR in this mouse model. These data show that RANTES (CCL5) is an important mediator of the pathophysiological responses seen in RSV infection. PMID- 12555674 TI - Dissecting TCR-MHC/peptide complex interactions with A2/peptide multimers incorporating tumor antigen peptide variants: crucial role of interaction kinetics on functional outcomes. AB - Soluble peptide/MHC-class-I (pMHC) multimers have recently emerged as unique reagents for the study of specific interactions between the pMHC complex and the TCR. Here, we assessed the relative binding efficiency of a panel of multimers incorporating single-alanine-substituted variants of the tumor-antigen-derived peptide MAGE-A10(254-262) to specific CTL clones displaying different functional avidity. For each individual clone, the efficiency of binding of multimers incorporating MAGE-A10 peptide variants was, in most cases, in good although not linear correlation with the avidity of recognition of the corresponding variant. In addition, we observed two types of discrepancies between efficiency of recognition and multimer binding. First, for some peptide variants, efficient multimer binding was detected in the absence of measurable effector functions. Some of these peptide variants displayed antagonist activity. Second, when comparing different clones we found clear discrepancies between the dose of peptide required to obtain half-maximal lysis in CTL assays and the binding efficiency of the corresponding multimers. These discrepancies, however, were resolved when the differential stability of the TCR/pMHC complexes was determined. For individual clones, decreased recognition correlated with increased TCR/pMHC off-rate. TCR/pMHC complexes formed by antagonist ligands displayed off-rates faster than those of TCR/pMHC complexes formed with weak agonists. In addition, when comparing different clones, the efficiency of multimer staining correlated better with relative multimer off-rates than with half-maximal lysis values. Altogether, the data presented here reconcile and extend our previous results on the impact of the kinetics of interaction of TCR with pMHC complexes on multimer binding and underline the crucial role of TCR/pMHC off-rates for the functional outcome of such interactions. PMID- 12555675 TI - A single dose of oral DNA immunization delivered by attenuated Salmonella typhimurium down-regulates transgene expression in HBsAg transgenic mice. AB - The efficacy of immunization with Salmonella typhimurium aroA to deliver the plasmid pRc/CMV-HBsAg (i.e. an oral DNA vaccine) was compared with that of intramuscular immunization with the same plasmid DNA, and with recombinant HBsAg protein, in a HBsAg transgenic mouse model. A single dose of oral DNA vaccine evoked vigorous Th1 cell and CTL responses and production of IgG2 subclass of anti-HBs after 2 weeks, and this was accompanied by a transient hepatitic flare with elevated alanine aminotransferase in the first 3 weeks. Concomitantly, the level of HBsAg-mRNA in liver tissues decreased by more than fourfold and viral antigen expression was curtailed markedly in hepatocytes compared with controls. Hepatitic flare subsided after 3 weeks, but suppression of the transgene expression was continued in the absence of overt liver pathology for the remaining duration of the experiment (i.e. 12 weeks), and possibly beyond. The other vaccines could also break immune tolerance, but this was achieved only after repeated booster doses of the respective vaccines, and they did not affect transgene expression, or induce hepatic flare. We previously showed in non transgenic mice that immunization by the oral DNA vaccine is achieved by an active intestinal infection with a bacterial carrier that is an adept intracellular parasite, and the immune response to the vaccination is orchestrated by phagocytic APC. Our present findings further implicated that the combined effects of an innate and a specific immune response induced by oral DNA vaccination are crucial in down-regulating HBsAg-transgene expression in hepatocytes. PMID- 12555676 TI - Regulation of IgG antibody responses by epitope density and CD21-mediated costimulation. AB - Epitope density and organization have been shown to be important factors for B cell activation in many animal model systems. However, it has been difficult to separate the role of antigen organization from the role of local antigen concentrations because highly organized antigens are usually particulate whereas non-organized antigens are more soluble. Hence, highly organized and non organized antigens may interact with different cell types and in different locations within lymphoid organs. In order to assess the role of antigen organization in regulating B cell responses, we immunized mice with highly repetitive virus-like particles, which exhibit different epitope densities covalently attached to them. Therefore, the same particulate structure was used to present identical epitopes that differed in their degree of organization. Induction of epitope-specific IgM titers, reflecting early B cell activation, were unaffected by the degree of epitope density. Furthermore, the absence of Th cells or CD21/CD35 did not reduce the IgM response. In contrast, the degree of organization was a critical factor influencing the magnitude of the epitope specific IgG response. Moreover, the threshold for IgG responses was shifted in the absence of CD21/CD35, resulting in the requirement for higher epitope densities to allow efficient IgG responses. Thus, IgG but not IgM responses are regulated by epitope density and B cell costimulatory thresholds. PMID- 12555677 TI - CD4-deficient T helper cells are capable of supporting somatic hypermutation and affinity maturation of germinal center B cells. AB - Collaborative interactions between B lymphocytes and CD4+ helper T cells are necessary for the induction of Ab responses to most protein Ag and for the generation of memory B cells in germinal centers. To study the role of the CD4 molecule in the germinal center response and in the development of B cell memory, we have investigated T helper function in the initiation and maturation of humoral immunity in CD4-deficient mice. In the absence of CD4+ T cells, immunization with thymus-dependent Ag was able to induce germinal center formation and Ig somatic hypermutation. In addition, Ag-driven affinity maturation and development of B cell memory were largely intact in CD4-deficient mice. Thus, CD4-deficient T helper cells are able to collaborate with Ag activated B cells to elicit the germinal center reaction, switch on the mutational machinery, and deliver signals necessary for B cell memory development. PMID- 12555678 TI - Consequences of OX40-OX40 ligand interactions in langerhans cell function: enhanced contact hypersensitivity responses in OX40L-transgenic mice. AB - Langerhans cells (LC) represent the dominant antigen-presenting cells (APC) in the epidermis and thus play an important role in cutaneous immune responses to approaching pathogens. These responses are mediated by several costimulatory molecules after antigenic challenge. OX40 ligand (OX40L), a member of TNF superfamily, is expressed on several APC such as splenic dendritic cells (DC) and activated B cells. This molecule has been reported to provide potent costimulation in APC-T cell interactions upon binding to its cognate receptor, OX40, on activated T cells. Little is known, however, regarding OX40L expression and function on LC. In the present study, we report the expression of both OX40L and OX40 on differentiated LC derived from draining lymph nodes in the FITC sensitized mice. During contact hypersensitivity responses, OX40L-deficient mice demonstrated a significant reduction in both hapten-induced ear swelling and hapten-specific T cell responses despite intact migratory responses. Conversely, these responses were markedly increased in two different OX40L-transgenic strains with variations in OX40L overexpression. In the LC-induced MLR, OX40L-deficient and OX40L-overexpressing LC were capable of reducing and elevating the responses of allogeneic CD4+ T cells, respectively. Thus the requirement of OX40L during the antigen presentation function of LC in T cell priming is here demonstrated. PMID- 12555679 TI - Ethics and ACHE. The Ethics Task Force will ensure that the Code of Ethics continues to be relevant. PMID- 12555680 TI - Our aging population Implications for healthcare organizations. PMID- 12555681 TI - Collaborative strategies for acute and post-acute care providers. PMID- 12555683 TI - Launching local ACHE chapters. ACHE is implementing a chapter structure to meet affiliates' needs. PMID- 12555682 TI - The specialty care debate: is there an answer? PMID- 12555684 TI - Maximizing the value of your facilities projects. Focusing on life-cycle costs will benefit your organization long-term. PMID- 12555685 TI - Your board and conflicts of interest. Taking appropriate measures encourages disclosure among board members. PMID- 12555686 TI - Education online. Gain knowledge, learn skills, or earn a degree via the Internet. PMID- 12555687 TI - Physicians and HIPAA compliance. Ensuring physician input can ease HIPAA preparation and implementation. PMID- 12555688 TI - Beyond the mentoring relationship. Create a learning plan to continue your professional development. PMID- 12555689 TI - Maximizing hospital capacity. Expediting patient throughput can provide an immediate solution to capacity shortages. PMID- 12555690 TI - Bridging the gap. An innovative program helps to combat racial disparities in healthcare. PMID- 12555691 TI - 2002 election results. How will the GOP's agenda affect healthcare spending. PMID- 12555692 TI - Meeting the needs of the chronically ill. Tampa General implements a care management model for the chronically ill. PMID- 12555693 TI - Uncovering the complexities of retroviral ribonuclease H reveals its potential as a therapeutic target. AB - Successful long-term management of HIV infection will require targeted inhibition of multiple steps essential for virus replication. Currently, both nucleoside- and nonnucleoside-based inhibitors of DNA polymerase function, in combination with antagonists of HIV protease, have been shown to be clinically beneficial. However, it is clear that RNase H activity of the multifunctional HIV-1 reverse transcriptase (RT) is absolutely required for completion of retroviral DNA synthesis, thereby rendering this function an attractive target for drug development. Although generally viewed as a sequence-independent activity, highly precise RNase H cleavage is required in order to remove the RNA primers of (-) and (+) strand DNA synthesis (a host-derived tRNA and the polypurine tract, respectively), thereby preserving the ends of linear DNA and facilitating integration. The availability of highly purified, recombinant RT/RNase H has allowed a thorough dissection of these multiple events and their potential for therapeutic intervention. Our current understanding of retroviral RNase H function and the status of small molecule inhibitors are the focus of this review. PMID- 12555694 TI - HIV-1 genetic recombination: experimental approaches and observations. AB - Many HIV-1 isolates contain interwoven genomic sections derived from multiple parental strains. Such chimeric genomes arise via genetic recombination. This review summarizes experimental approaches for addressing the frequency of HIV-1 genetic recombination during single cycles of viral replication in vitro, and describes factors--such as variation in extents of sequence homology and the metabolic state of the infected cell--that modulate recombination. Findings from such studies suggest that recombinogenic template switching is an even more common occurrence during HIV-1 DNA synthesis than is the introduction of base substitution errors. This implies that recombination is an inherent property of retroviral DNA synthesis, and that the vast majority of HIV-1 DNAs are biochemical recombinants. PMID- 12555695 TI - Street and network sampling in evaluation studies of HIV risk-reduction interventions. AB - Although sampling is a crucial component of research methodology, it has received little attention in intervention research with populations at risk for HIV infection. We review the challenges involved in sampling these populations for evaluating behavioral and social interventions. We assess the four strategies used for street and network sampling that have been reported in the HIV intervention research literature and used because traditional probability sampling was not possible. The sampling strategies are: 1) targeted, 2) stratified, (3) time-space, and (4) respondent-driven. Although each has strengths and limitations in terms of its ability to produce valid results that enhance generalizability, the choice of a particular strategy depends on the goal of the study, characteristics of the target population, and the availability of resources and time for collecting and analyzing sampling-related data. Continued efforts are needed to improve the sampling strategies used in evaluation studies of HIV risk-reduction interventions. PMID- 12555696 TI - Might the M184V substitution in HIV-1 RT confer clinical benefit? AB - The M184V substitution in HIV-1 RT develops rapidly following initiation of therapy with 3TC and confers high-level phenotypic resistance to this drug both in vitro and in vivo. Interestingly, the presence of M184V is also associated with alteration of several mechanisms relating to RT function that include decreased RT processivity, reduced nucleotide-dependent primer unblocking, increased fidelity, hypersensitization to other NRTIs, impaired viral fitness, and delayed appearance of mutations in RT that are responsible for resistance to thymidine analogues (i.e. thymidine-associated mutations or TAMs). Collectively, these factors might explain the residual antiviral effect and clinical benefit observed with continued use of 3TC in combination therapy regimens following the emergence of M184V. Indeed, the results of numerous controlled as well as observational clinical studies are suggestive of improved therapeutic outcome associated with continued usage of 3TC and maintenance of the M184V mutation. However, several of these trials did not possess adequate statistical power to resolve whether or not continued use of 3TC provided actual benefit, nor were they specifically designed to test the M184V benefit hypothesis in prospective fashion. There is a need for randomized clinical trials of this type in order to validate the potential benefit of maintenance of M184V and whether continued use of 3TC is the only means of attaining this objective. PMID- 12555697 TI - Simplification of antiretroviral treatment--how to sustain success, reduce toxicity and ensure adherence avoiding PI use. AB - Shortly after protease inhibitors (PI) began to be widely prescribed for the treatment of HIV infection in 1996, we learned that the virological efficacy of highly active antiretroviral therapy (HAART) was frequently associated with severe quality of life impairment. In fact, failure of PI-based regimens is often due to poor adherence resulting from high pill burden, complex dosing schedules, adverse events, or long-term toxicity. The metabolic and morphologic abnormalities associated with prolonged PI use are of particular concern: fat accumulation and abnormalities of lipid and glucose metabolism may increase the risk of cardiovascular disease, and body-shape changes (lipodystrophy) may also have a psychological impact or decrease patients' willingness to adhere to therapy. The efficacy of PI-sparing regimens in drug-naive subjects, which typically consist of nucleoside reverse transcriptase inhibitors (NRTIs) with or without non-nucleoside reverse transcriptase inhibitors (NNRTIs), gives support to simplification strategies. In patients who have maintained durable virologic suppression on a PI-based regimen, the replacement of a PI by an NRTI, usually abacavir (ABC), or an NNRTI, has been shown to be associated with maintenance of virologic suppression, while leading to improvements in adherence and in some PI related toxicities. The replacement of a PI by an NNRTI or ABC is usually followed by reductions in total cholesterol and triglycerides, though this is less true with efavirenz (EFV) than with nevirapine (NVP) or ABC. While there may be some improvement in fat accumulation after switching from a PI-based regimen, no improvement in lipoatrophy has been demonstrated after PI switches, possibly because this toxicity is due to NRTIs rather than PIs. However, NVP should be used with caution in patients co-infected patients with hepatitis B or C, and EFV may be poorly tolerated in individuals with psychiatric morbidity. Patients taking methadone may require dosage increases with switches to either NVP or EFV. PMID- 12555698 TI - [Settling of accounts according to diagnosis-related groups-- or "do we get enough money for our payments"?]. PMID- 12555699 TI - [Diagnosis and procedure coding in relation to the DRG system]. AB - INTRODUCTION: The aim was to investigate the consequences of missing or wrong diagnoses and procedure codes in relation to the DRG system. MATERIAL AND METHODS: All patients admitted to the orthopaedic department during the course of one week, 155 patients, were consecutively entered. Former diagnoses were registered from interviews with all the patients, former case notes, and present hospital records. They were then compared to the department case notes, including diagnosis and procedure codes. All codes were then compared in Visual DRG (version 97) for grouping. RESULTS: The coding was correct in 103 of 155 cases (65%). In 52 cases (35%) the coding was incorrect or insufficient, in 18 of the 52 cases (12% overall) it lead to a decrease in the DRG value, which extrapolated on a yearly base, would lead to a loss of DDK 23 million. In total, coding was incorrect or insufficient in one third of the records. DISCUSSION: Irrespective of whether the DRG system is implemented or not, it is important that departments register the correct diagnoses and procedures, not only those relevant to the department. There is a continued need to teach and inform the staff about the correct coding procedures. PMID- 12555700 TI - [The dysplastic hip I. Etiology, epidemiology and diagnosis]. AB - Magnetic resonance scanning, computer tomography, and dynamic ultrasound-scans have recently unveiled a broad spectrum of morphological anomalies of the dysplastic hip. The possibility of individualized treatment of late diagnosed and residual dysplasia of the hip in childhood and adulthood has renewed the interest in this orthopaedic topic. PMID- 12555701 TI - [The dysplastic hip II. Treatment]. AB - The possibility of individualized treatment of residual dysplasia of the hip in childhood and adulthood has renewed the interest in this orthopaedic field and new procedures of treatment have been devised. This article reviews current knowledge of the treatment of the dysplastic hip. PMID- 12555702 TI - [Radiotherapy of primary breast cancer. An analysis of a systematic Cochrane review]. PMID- 12555703 TI - [Prevention of colorectal cancer in families with hereditary nonpolyposis colorectal cancer]. AB - INTRODUCTION: Hereditary non-polyposis colorectal cancer (HNPCC) is a dominant inherited colorectal cancer syndrome, with lifetime risk up to 90% of developing colorectal cancer (CRC) for carriers of the genes. Screening with colonoscopy can reduce the CRC-rate by 62% and prevent CRC-deaths. The HNPCC-Register was established with the aim of identification and registration of Danish HNPCC families and coordination of surveillance. MATERIALS AND METHODS: The results of 703 colonic screenings in 396 asymptomatic individuals from 150 HNPCC-families were analysed and related to the family diagnosis. RESULTS: In 112 asymptomatic individuals, 31, colorectal cancers and 140 adenomas were diagnosed. Neoplasia was found in one fifth of the examinations and almost one third of the individuals developed an asymptomatic neoplasia which was detected by screening. At the time of the diagnosis, 77% of the colorectal cancers were localized (Dukes stage A or B). DISCUSSION: In Finland, screening for colorectal cancers in HNPCC families prevents 25-45-fold more CRC-deaths compared to general population screening for colorectal cancer. The results of the Danish colonic HNPCC screening reveal neoplasia in 21% of the examinations and the colorectal cancers found by screening were at a more favourable Dukes stage compared to sporadic CRC, and probably also CRC found by general population screening. The HNPCC Register initiates surveillance together with the clinical genetic departments, but the results of the recommended colonic examinations are not automatically reported to the HNPCC-Register. To enable an in-depth investigation of the effect of CRC-screening in all Danish HNPCC-families, further financial resources should be allocated to the HNPCC-Register to collect the results. PMID- 12555704 TI - [Factors of importance for early and late admission of patients with stroke and transient cerebral ischemia]. AB - INTRODUCTION: Early admission after stroke and TIA is important in modern stroke treatment. We studied the time delay to admission and explored predictive factors of early/late admission. MATERIAL AND METHODS: The study was prospective and community-based comprising all patients with stroke or TIA admitted to a Copenhagen hospital from 1 September 1999 to 30 April 2000. The catchment area is well defined with 283,000 inhabitants. All had a neurological examination and a structured interview within three days with registration of age, gender, premorbid Rankin, Scandinavian stroke scale score, time of onset, knowledge of the cause of symptoms, cohabitation, alone at onset, whether admitted by a general practitioner (GP) or by ambulance after calling emergency, and relevant stroke risk factors. Univariate and multivariate statistics were used. RESULTS: Altogether 494 patients with stroke and 63 with TIA were entered; 49% were admitted by a GP, 38% by ambulance after calling emergency, 13% via other routes. Time from onset to hospital admission could be assessed reliably in 374 patients (67%) and was a median of 2.6 hours; 37% arrived within 0-3 hours, 55% within 0-6 hours. Patients calling an ambulance over emergency arrived at a median of 1.0 hour after the stroke; those calling the GP a median of 6.0 hours after the stroke. In a multivariate analysis only admission by ambulance after calling emergency (OR 5.7), TIA (OR 5.6), or the patient's knowledge of the cause of the symptoms (OR 2.2) were predictors of early admission. DISCUSSION: Patients with stroke or TIA in a Danish metropolitan area arrive at hospital a median of 2.6 hours after the stroke. Admission by ambulance after calling emergency was associated with the shortest onset to admission time. PMID- 12555705 TI - [Junior physicians' acquisition of theoretical knowledge at a surgical department. Evaluation in the form of multiple-choice tests]. AB - INTRODUCTION: Denmark has no tradition of formal assessment of junior doctors' theoretical knowledge. We therefore wanted to investigate whether the knowledge of surgical junior doctors has actually increased. MATERIAL AND METHODS: Twenty three junior doctors were confronted with the same multiple-choice questionnaires twice during a six months period. Subsequently, they were asked whether they would approve the implementation of multiple-choice questionnaires and formal assessment (exam) in the Danish specialist training in surgery. RESULTS: The participants got a significantly higher score the second time they were confronted with the multiple-choice questionnaires (p = 0.0003). When dividing the participants into two groups, viz. young vs. old junior doctors, it was observed that the latter group had a higher score overall but augmented their scores less in relation to their young colleagues (p = 0.038). Forty percent of the participants replied that the multiple-choice questionnaires had motivated them to do further theoretical studies, and half of them approved the introduction of an exam in surgery in the Danish specialist training. DISCUSSION: The theoretical knowledge of junior doctors does actually increase in a relatively short period of time during their employment at a department of surgery. As expected the old junior doctors got a higher score compared to their young colleagues. However, the latter group augments its knowledge relatively more. Multiple-choice questionnaires are easy, accessible, and inexpensive tools for evaluation as well as stimulation for doing further theroretical studies. PMID- 12555706 TI - [Lung function changes during anesthesia and thoracic surgery]. AB - After anaesthesia and thorax surgery, a significant reduction in pulmonary function with up to a 50% decrease in FEV1, FVC, and FRC is seen, leading to a high risk of atelectasis and hypoxia, and therefore making respiratory complications the major course of perioperative morbidity and mortality in this group of patients. This severe reduction in lung function gradually diminishes within three weeks, but postoperative pain and sedation increase the deterioration in lung function, and treatment is therefore based on anaesthetic drugs with a short elimination time and effective postoperative pain treatment with epidural analgesia or other regional blockade, and minimal use of opioids. Together with more sparing surgical methods, it is possible to operate on patients with severely reduced preoperative lung function (FEV1 = 0.651 or 22% of predicted value) with lung resection and immediate extubation. A reduction in pulmonary complications after thoracic surgery from 25-30% down to 10-15% is also seen, depending on age and preoperative lung function. PMID- 12555707 TI - [Excess mortality associated with antibiotic resistant Salmonella typhimurium]. AB - INTRODUCTION: The use of antimicrobials in food production has been associated with drug-resistance in foodborne pathogens such as Salmonella and Campylobacter. However, little is known about the health impact of antimicrobial drug-resistance in these pathogens. In a matched cohort study, we determined the mortality associated with drug-resistance in Salmonella Typhimurium. MATERIAL AND METHODS: Data from the Danish Surveillance Registry for Enteric Pathogens were linked with the Civil Registration System and the Danish National Patient Registry. By survival analysis, the two-year mortality among patients was compared with a matched sample of the general Danish population adjusted for differences in comorbidity. RESULTS: Among 2047 cases of S. Typhimurium, 59 deaths were identified. Patients with pan-susceptible strains of S. Typhimurium were 2.3 times (95 per cent confidence interval [CI] 1.5-3.5) more likely to die in the two-year follow-up period after infection than persons in the general Danish population. Patients infected with strains resistant to ampicillin, chloramphenicol, streptomycin, sulfonamide, and tetracycline were 4.8 times (95 per cent CI 2.2-10.5) more likely to die, whereas quinolone-resistance was associated with a 10.3 times (95 percent CI 2.8-37.8) higher mortality than the general population. DISCUSSION: Overall, infections with S. Typhimurium were associated with excess mortality. Patients infected with multi-drug resistant strains had a tendency towards higher mortality than patients infected with non multiresistant strains. Particular risk was associated with quinolone-resistance in foodborne S. Typhimurium, and the excess mortality among patients infected with multi-drug resistant S. Typhimurium could be attributed to additional quinolone resistance in many of the multi-drug resistant isolates. PMID- 12555708 TI - [Fatal poisoning with Letigen]. AB - Two cases of fatal caffeine and ephedrine intoxication due to ingestion of an anorectic drug Letigen (200 mg caffeine and 20 mg ephedrine) are reported. A 39 year-old female with a history of prior suicide attempts was found dead in her home by her husband. An empty bottle of Letigen was found. The other case was a 31-year-old female who was found dead in her car which had stopped on the motorway. She had been vomiting in the car. The toxicological analysis revealed fatal ephedrine and caffeine intoxication in both cases. In the first case the manner of death was suicide, the other death remained unresolved. One fatal Letigen intoxication due to suicidal ingestion in Denmark has previously been published. PMID- 12555709 TI - [Laparoscopic treatment of a benign hepatic cyst]. AB - We present a case of laparoscopic treatment of a symptomatic solitary hepatic cyst and the use of preoperative laparoscopic ultrasound scanning and ultrasonic scalpel. PMID- 12555710 TI - [Isolated ventricular "non-compaction". A rare form of cardiomyopathy]. AB - We describe cardiomyopathy, isolated ventricular non-compaction, in a 21-year old woman. PMID- 12555711 TI - [Dexamethasone to adults with acute bacterial meningitis]. PMID- 12555712 TI - ["Status Cancer" 2]. PMID- 12555713 TI - [Children with hypernatremia and weight loss]. PMID- 12555714 TI - [DSAM, diabetes and hypertension]. PMID- 12555715 TI - [Is Letigen really SO dangerous?]. PMID- 12555716 TI - Lessons learned provide practical ideas for improving patient safety. PMID- 12555717 TI - New alcohol-based hand rubs catch on quickly with staff, MDs. PMID- 12555718 TI - MIS knees are a hot topic in orthopedics. PMID- 12555719 TI - A high-level view of health care. PMID- 12555720 TI - Pressure ulcers: sorting out the evidence. PMID- 12555722 TI - The postop phone call: an effective tool? PMID- 12555721 TI - Thinking about eye instruments and CJD. PMID- 12555723 TI - 25 Health care trends. What's hot, what's not, and what does the future hold. AB - As we usher in 2003, America's health care system remains in a chaotic state. Will managed care live or die? Will quality improvement efforts pay off? Are we ready for the next bioterrorism attack? Will the shortage of physician soon rival the shortage of nurses? To help gauge where health care stands today and what the future holds, The Physician Executive asked doctors who serve on ACPE's peer review panel to list the hottest health care trends in the U.S right now. Then, we took the list to three respected health care futurists -- Leland Kaiser, PhD, Jeff Goldsmith, PhD, and Russel Coile, MBA -- and asked them for their insights on the trends. Yes, Kaiser, Goldsmith and Coile are opinionated. Yes, they're controversial. But no matter whether you agree or disagree with their views, the three health care futurists' comments could spark discussions that will help shape U.S. health care this year and beyond. The trends are presented in no particular order. PMID- 12555724 TI - Teaching clinic lowers pharmacy costs. How to achieve more cost-effective prescribing. PMID- 12555725 TI - Pros and cons of prescription drug benefit changes. AB - Check out a scorecard of ideas for improving prescription drug programs. Look at the programs' effects on patients, physicians and pharmaceutical companies. PMID- 12555726 TI - Is health care a right or a privilege? AB - In this overview of the American health system, the author make arguments for declaring health care a right in the U.S. Constitution. Learn why he believes this is necessary. PMID- 12555727 TI - The competitive advantage: strategic thinking for physician leaders. AB - Physician executives should manage their career strategies like a well-run company. Learn how to start thinking aout your career strategy. PMID- 12555728 TI - Evaluating risk: global fees and episodic care. AB - In 1999, two articles in The Physician Executive -- "Part I: Global Theory and the Nature of Risk (July-August)." and "Part II: Towards a Choice-Based Model of Managed Care (October-November)" -- outlined the flaws of orthodox managed care theory and highlighted the unique advantages of moving to a genuinely market based model, which included the concept of direct contracting for integrated episodes of care. This follow-up focuses on comparing an episode contracting system to a traditional capitated program and outlines the features that make this approach much more attractive to physicians, payers, and most importantly patients. PMID- 12555729 TI - Don't overreact to Enron. Greed isn't a good choice, but is altruism any better? PMID- 12555730 TI - 8 traits of the perpetually employed person. PMID- 12555731 TI - Developing a marketing strategy. PMID- 12555732 TI - Getting up to bat. PMID- 12555733 TI - Who will care for the aging American population? PMID- 12555734 TI - Lessons learned from service to the medical licensing board. PMID- 12555735 TI - The 100-year-old patient. What we can learn about aging from centenarians. PMID- 12555736 TI - Can thrombophilia testing help to prevent recurrent VTE? Part 2. PMID- 12555737 TI - The challenge of childhood depression and ADHD. PMID- 12555738 TI - An Afghan journal. PMID- 12555739 TI - Treatment-resistant scabies and lice infections. PMID- 12555740 TI - Recurrent pustules on a baby's hands. PMID- 12555741 TI - [Statins and cardiovascular disease. A changing understanding, a reinforced concept]. PMID- 12555742 TI - Development of polyamine analogs as cancer therapeutic agents. AB - Natural polyamines (putrescine, spermidine, and spermine) are aliphatic cations with multiple functions in cell growth and differentiation. Alterations in the polyamine structure provide a strategy to synthesize analogs that can interfere with the cellular functions of natural polyamines. Analogs of spermine are particularly effective in modifying the synthesis, catabolism, and uptake of natural polyamines. The increased requirement of natural polyamines in cancer cell growth makes it possible to utilize the polyamine pathway as a therapeutic target in cancer cells. Because polyamine functions extend from membrane phospholipid structure and signal transduction to DNA structure and conformational transitions, it is likely that the action of polyamine analogs also permeates to these sites of polyamine action. For the same reason, toxicity of polyamine analogs might be considerable. However, it is possible to design polyamine analogs that target a specific function of polyamines in cancer cells, thereby enhancing selectivity for inducing apoptosis of cancer cells. Alternatively, polyamine analogs may potentiate the action of other anticancer agents and become an effective tool in cancer chemotherapy. In either case, further research into the action of polyamine analogs will open up new opportunities in the fight against cancer. PMID- 12555743 TI - [State-tolerated nicotine addiction in conscripts]. PMID- 12555744 TI - To: Pundziute - Lycka. A et al. (2002). The incidence of type 1 diabetes has not increased but shifted to a younger age at diagnosis in the 0-34 years group in Sweden 1983-1998. Diabetologia 45: 783-791. PMID- 12555745 TI - Comparison of alterations in insulin signalling pathway in adipocytes from Type II diabetic pregnant women with gestational diabetes mellitus. PMID- 12555746 TI - The 'reference trap' revisited: examples of the dangers in using ratios to describe fetoplacental angiogenesis and trophoblast turnover. AB - Examples of the use of relative or ratio data are to be found throughout biomedical sciences and include such variables as stereological component densities per cell volume (morphology), transport rates per tissue volume or membrane surface (physiology), gold labelling frequencies (immunocytochemistry) and amounts of DNA relative to protein (biochemistry). This report emphasizes the potential pifalls associated with drawing biological conclusions and interpretations from relative data when there has been no attempt to monitor the absolute size of the reference space. This is know as the 'reference trap'. The dangers to interpretation inherent in confining results to such data are illustrated using the sorts of data found in current placental studies on fetoplacental angiogenesis and villous trophoblast turnover but are pertinent to many other areas. Where it is possible to do so, these dangers can be avoided by the simple expedient of estimating the size of the pertinent reference space and using this to calculate absolute values. Sometimes (e.g. when relying on biopsy samples), the size of the reference cannot be determined. In such cases, ratio data must be interpreted with due caution. PMID- 12555748 TI - [A case report of an elderly diabetic patient with complication of multiple organ tuberculosis]. PMID- 12555749 TI - [Double uterus and hydatidiform mole in the malformed uterus: report of one case]. PMID- 12555750 TI - [Transfusion of fresh coagulated plasma: products, indications. General methods and recommendations]. PMID- 12555751 TI - [The summer seminar by Kansai Society of Circulatory Technology]. PMID- 12555752 TI - My patient's been poisoned-what are my responsibilities? PMID- 12555753 TI - [Transfusion of erythrocyte substitutes: products, indications, alternatives. General methods and recommendations]. PMID- 12555754 TI - Understanding neuropathic pain. PMID- 12555755 TI - [Studies on automated detection methods and related techniques for mammographic masses]. PMID- 12555756 TI - Using a BNP test to identify heart failure. PMID- 12555757 TI - How to wrap an above-the-knee amputation stump. PMID- 12555758 TI - Lab workers: small group, big risk. PMID- 12555759 TI - Documenting surgical incision site care. PMID- 12555760 TI - Understanding echinacea. PMID- 12555761 TI - Protecting your patient's aging skin. PMID- 12555762 TI - The production of emotional prosody in varying degrees of severity of apraxia of speech. AB - The purpose of this study was to examine the abilities of subjects with varying degrees of apraxia of speech (AOS) to produce emotional prosody. One subject with mild AOS, one subject with moderate AOS and a control subject were asked to produce sentences with happy, sad and neutral emotional intent in a repetition and a reading experiment and naive listeners were asked to identify the intended emotion of the productions based upon the prosodic structures. Acoustic analyses of the subjects' productions revealed that unlike the control subject, the AOS subjects did not produce differences in FO, duration and amplitude cues within their prosodic structures to convey the different emotions. Naive listeners were able to identify the emotional intent of the control subject's productions but were unable to distinguish between the utterances of the experimental subjects. These findings are discussed in conjunction with suggestions for future research in this area. LEARNING OUTCOMES: The reader will (1) be able to define apraxia of speech and identify speech characteristics associated with apraxia of speech; (2) acquire an understanding of prosodic cues used to signal emotional intent; and (3) develop an understanding of the relationship between apraxia of speech and the production of emotional prosody. PMID- 12555763 TI - Infection control related sentinel events. PMID- 12555764 TI - Federalism: how the principle works on state and local levels. PMID- 12555765 TI - The unfair playing field. PMID- 12555766 TI - Coronary symptoms. PMID- 12555767 TI - Parental responsibility. PMID- 12555768 TI - Panoramic radiography. PMID- 12555769 TI - Dental student motivations. PMID- 12555771 TI - ["Aggressive shock therapy": a relic?]. PMID- 12555770 TI - Creating meaning and value in substance abuse education. AB - Medical faculty committed to teaching about substance abuse face the challenges of teaching about a stigmatized disease. They often work with little support and few colleagues. These clinician educators must find tools to help sustain them in this work. Knowledge of emerging data on effective diagnosis and treatment of substance abuse in the primary care setting and excellent clinical skills in managing patients with substance abuse are key. More important is the ability to find sustenance and inspiration in working with the addicted patients who have been our teachers. PMID- 12555772 TI - [Perioperative metformin and lactacidosis]. PMID- 12555773 TI - [Anesthesia in allergic diathesis. Allergic diathesis, typical triggers in the perioperative phase, perioperative prophylaxis]. PMID- 12555774 TI - Divergence in mate choice systems: does evolution play by rules? AB - Understanding the genetic bases of phenotypes associated with the earliest stages of divergence will reveal a great deal about species formation. I review a number of model systems, most involving plant-insect interactions, that have already revealed genetic aspects of incipient speciation. It is suggested that progress in understanding the causal forces driving mating signal evolution and incipient speciation will be expedited in model systems where; (1) ecological and evolutionary information is available, (2) different aspects of mating behaviors that function in mate and/or species recognition are known, (3) genetic analysis of single phenotypes is undertaken, (4) analysis of sexual selection and isolation is performed under natural conditions (or in the wild), and (5) comparative data from related species are available to assess phylogenetic trends. PMID- 12555775 TI - Recombination and the divergence of hybridizing species. AB - The interplay between hybridization and recombination can have a dramatic effect on the likelihood of speciation or persistence of incompletely isolated species. Many models have suggested recombination can oppose speciation, and several recent empirical investigations suggest that reductions in recombination between various components of reproductive isolation and/or adaptation can allow species to persist in the presence of gene flow. In this article, we discuss these ideas in relation to speciation models, phylogenetic analyses, and species concepts. In particular, we revisit genetic architectures and population mechanisms that create genetic correlations and facilitate divergence in the face of gene flow. Linkage among genes contributing to adaptation or reproductive isolation due to chromosomal rearrangements as well as pleiotropy or proximity of loci can greatly increase the odds of species divergence or persistence. Finally, we recommend recombination to be a focus of inquiry when studying the origins of biological diversity. PMID- 12555776 TI - The evolution of hybrid infertility: perpetual coevolution between gender specific and sexually antagonistic genes. AB - A new hypothesis is proposed for the rapid evolution of postzygotic reproductive isolation via hybrid infertility. The hypothesis is motivated by two lines of experimental research from Drosophila melanogaster that demonstrate that sexually antagonistic fitness variation is abundant and that epistatic fitness variation on the Y chromosome is common. The hypothesis states that the expression of sexually antagonistic genes leads to a 'gender-load' in each sex. In response, gender-limited reproductive genes are selected to ameliorate, through pleiotropy, the expression of sexually antagonistic genes. Chronic coevolution between gender limited genes and gender-unlimited sexually antagonistic genes causes rapid divergence of reproductive proteins among allopatric populations, ultimately leading to hybrid infertility. PMID- 12555777 TI - Modelling the evolution of female choice strategies under inbreeding conditions. AB - Recently, many mate choice studies have discussed the role of genetic compatibility and inbreeding for the evolution of mate choice. With population genetic simulations I compared the potential advantage of three different female choice strategies under inbreeding conditions. Females were assumed to benefit indirectly via a preference for (i) complementary males, (ii) males with few detrimental mutations, and (iii) non-inbred males. Probably related to the reduced inbreeding depression in offspring of choosy females, the choice-allele increased for all three strategies. However, the advantage of the strategies differed widely. Choice of males with fewer mutations provided a comparatively large advantage, choice of complementary males led to a reasonable advantage, and choice of non-inbred males only resulted in a minor advantage of female choice. My results show that complementary mate choice can be almost as beneficial as conventional good-genes choice of mates with lower genetic load. Compared to the two other mate choice strategies, choice of non-inbred males is less likely to contribute to the evolution of costly mate choice. The results of a recent study showing that female sticklebacks prefer males with a larger number of MHC-loci is thus unlikely to be related to an indirect benefit of choosing non-inbred males. PMID- 12555778 TI - Single gene mutations in Drosophila: what can they tell us about the evolution of sexual behaviour? AB - The molecular analysis of specific mutant genes that affect the courtship behaviours of Drosophila melanogaster males and females is discussed in the light of the possibility that they may contribute to mate choice. There is clear evidence that some genes can act as a reservoir of species-specific behaviour, particularly for the male actions during courtship. However, to date there has not been a single genetic locus that has been isolated at the molecular level and shown to be associated with a change in female preference. There are some promising avenues of exploration, in that recent genetic analyses suggest that a small number of genes may make major contributions to female preferences. Finally a candidate gene approach is advocated in which orthologous genes from other species of Drosophila are used as 'natural' mutations, and transformed into D. melanogaster hosts to investigate whether they carry species-specific mating information of the donor. PMID- 12555779 TI - Sexual signaling and speciation, a microevolutionary perspective. AB - Despite the growing evidence that sexual selection can drive speciation, the evolution of sexual signals in natural populations is far from being well understood. Sexual signals evolve in response to a variety of factors. Some of the most important selective factors are conspecifics, transmission efficiency in a particular environment, detection by predators, and phylogenetic constraints. These factors have been addressed quite successfully in studies of single types of signals in both vertebrates and invertebrates. However, it is less clear how multimodal signals evolve because the factors listed above will act on every component of the signaling system, and the relative weights of each type of signal must be taken into account. Species of Drosophila are excellent for such analyses because they are amenable to both phenotypic experimentation and genetic manipulation. This paper presents an approach that involves two analyses: studies of which signals are sexually selected within a species, and parallel studies of the signals that are involved in behavioral isolation between closely related species. If the same signal characteristics are involved in both processes, they would provide support for the hypothesis that sexual selection can drive speciation. This approach is illustrated with studies of Hawaiian Drosophila and a review of signals that could be sexually selected in Drosophila melanogaster. PMID- 12555780 TI - Reproductive isolation in natural populations of Drosophila melanogaster from Brazzaville (Congo). AB - The aim of this work is to analyze the homogamy previously detected between two natural populations of Drosophila melanogaster from Brazzaville. It is shown that mating isolation was still maintained under laboratory conditions 10 years after the populations samples were trapped. Isolation seemed to be due mainly to premating isolation and we checked for any suggestion of post-mating mortality of hybrids. Pre-mating isolation was not symmetrical, and significant chi2 values were found in 3/4 possible 3-way mating choice experiments. The only exception involved a male from the countryside and two females (one from each population) for which no significant mating preference was detected. Mortality of hybrids was intermediate between those of the parental strains showing a clear maternal effect and the existence of partial dominance. Major differences in the cuticular hydrocarbons were also found and they could account for the isolation. These findings in populations from African breweries indicate that they are closely related to European ones, suggesting that this phenomenon is not a case of sympatric speciation, but probably attributable to the reintroduction of an allopatric population. PMID- 12555781 TI - Genetics of sexual isolation based on courtship song between two sympatric species: Drosophila ananassae and D. pallidosa. AB - Sexual isolation has been considered one of the primary causes of speciation and its genetic study has the potential to reveal the genetics of speciation. In Drosophila, the importance of courtship songs in sexual isolation between closely related species has been well investigated, but studies analysing the genetic basis of the difference in the courtship songs associated with sexual isolation are less well documented. Drosophila ananassae and Drosophila pallidosa are useful for studies of sexual isolation, because of their sympatric distribution and absence of postmating isolation. Courtship songs are known to play a crucial role in sexual isolation between these two species, and the female discrimination behaviour against the courting male has been revealed to be controlled by a very narrow region on the second chromosome. In this study we investigated the genetic basis controlling the song differences associated with their sexual isolation, using intact and wingless males with chromosomes substituted between species. The results obtained from F1 hybrid males between these species indicate the dominance of the song characters favoured by D. pallidosa females. In addition, the results obtained from backcross F2 males indicate that chromosome 2 had a major effect on the control of the song characters associated with sexual isolation. PMID- 12555782 TI - Evolutionary experiments on mate recognition in the Drosophila serrata species complex. AB - It is becoming increasingly apparent that at least some aspects of the evolution of mate recognition may be amenable to manipulation in evolutionary experiments. Quantitative genetic analyses that focus on the genetic consequences of evolutionary processes that result in mate recognition evolution may eventually provide an understanding of the genetic basis of the process of speciation. We review a series of experiments that have attempted to determine the genetic basis of the response to natural and sexual selection on mate recognition in the Drosophila serrata species complex. The genetic basis of mate recognition has been investigated at three levels: (1) between the species of D. serrata and D. birchii using interspecific hybrids, (2) between populations of D. serrata that are sympatric and allopatric with respect to D. birchii, and (3) within populations of D. serrata. These experiments suggest that it may be possible to use evolutionary experiments to observe important events such as the reinforcement of mate recognition, or the generation of the genetic associations that are central to many sexual selection models. PMID- 12555783 TI - Herbivorous insects: model systems for the comparative study of speciation ecology. AB - Does ecological divergence drive species-level evolutionary diversification? How so and to what degree? These questions were central to the thinking of the evolutionary synthesis. Only recently, however, has the ecology of speciation become an important focus of empirical study. Here, we argue that ecologically specialized, phylogenetically diverse, and experimentally tractable herbivorous insect taxa offer great opportunities to study the myriad mechanisms by which ecology may cause reproductive isolation and promote speciation. We call for the development and integrated experimental study of a taxonomic diversity of herbivore model systems and discuss the availability and recent evaluation of suitable taxa. Most importantly, we describe a general comparative framework that can be used to rigorously test a variety of hypotheses about the relative contributions and the macroevolutionary generality of particular mechanisms. Finally, we illustrate important issues for the experimental analysis of speciation ecology by demonstrating the consequences of specialized host associations for ecological divergence and premating isolation in Neochlamisus bebbianae leaf beetles. PMID- 12555784 TI - The inheritance of mating songs in two cryptic, sibling lacewing species (Neuroptera: Chrysopidae: Chrysoperla). AB - Speciation often involves incremental responses to natural selection and results in large scale genomic changes, but it may also occur abruptly and with little genetic imprint, as seen in some complexes of cryptic species. Recent attention has focused on sexual selection in rapid speciation, because it can disrupt premating signals that mediate reproductive isolation. Some models require that environmental adaptation assist sexual selection during speciation, while others show that populations can diverge through mate choice alone. We propose that speciation involving environmental adaptation in premating signals is likely to have a polygenic basis, while speciation due to arbitrary changes in premating signals could be accompanied by changes at just a few loci. The sibling species Chrysoperla plorabunda and Chrysoperla johnsoni belong to a large complex of cryptic species of green lacewings, and meet all criteria for speciation via sexual selection. We perform a genetic analysis of line crosses between them, focusing on their substrate-borne premating songs. Measurements of seven song features and their principal components are compared among parentals, F1 and F2 hybrids, and backcrosses. The distributions of phenotypes are consistent with a model of more than one, but not many, genes. Sex linkage and/or maternal effects are negligible. C. plorabunda is dominant for most traits affecting mate choice. Bayesian analyses of segregation variance show significant additive and epistatic effects on line means. A Bayesian Castle-Wright estimate suggests that relatively few effective factors are responsible for variation in volley period (x = two factors), number of volleys per song (x = 1), and PC-1 (x = 4). Our results are consistent with simple genetic architecture of songs, supporting a role for major genes in premating isolation and strengthening the notion that mate choice alone, without significant environmental adaptation, has been responsible for generating new lacewing species. PMID- 12555785 TI - Divergence and reproductive isolation in the early stages of speciation. AB - To understand speciation we need to identify the factors causing divergence between natural populations. The traditional approach to gaining such insights has been to focus on a particular theory and ask whether observed patterns of reproductive isolation between populations or species are consistent with the hypothesis in question. However, such studies are few and they do not allow us to compare between hypotheses, so often we cannot determine the relative contribution to divergence of different potential factors. Here, I describe a study of patterns of phenotypic divergence and premating and postmating isolation between populations of the grasshopper Chorthippus parallelus. Information on the phylogeographic relationships of the populations means that a priori predictions from existing hypotheses for the evolution of reproductive isolation can be compared with observations. I assess the relative contributions to premating isolation, postmating isolation and phenotypic divergence of long periods of allopatry, adaptation to different environments and processes associated with colonisation (such as population bottlenecks). Likelihood analysis reveals that long periods of allopatry in glacial refugia are associated with postmating reproductive isolation, but not premating isolation, which is more strongly associated with colonisation. Neither premating nor postmating isolation is higher between populations differing in potential environmental selection pressures. There are only weak correlations between patterns of genetic divergence and phenotypic divergence and no correlation between premating and postmating isolation. This suggests that the potential of a taxon to exercise mate choice may affect the types of factor that promote speciation in that group. I discuss the advantages and disadvantages of the general approach of simultaneously testing competing hypotheses for the evolution of reproductive isolation. PMID- 12555786 TI - Genetic and behavioral components of the cryptic species boundary between Laupala cerasina and L. kohalensis (Orthoptera: Gryllidae). AB - Cryptic species are often hypothesized on the basis of differences in courtship signals. These signal differences suggest that mate recognition systems, which include both courtship signals and responses to those signals, have diverged between genetically isolated populations. Cryptic species are therefore thought to represent distinct genetic units, the boundaries of which are maintained by premating incompatibilities, specifically by receiver preferences for conspecific signals. Laupala cerasina and L. kohalensis are sympatric species of swordtail crickets endemic to the big island of Hawaii, that are distinguishable by differences in male courtship song. We first tested whether groupings hypothesized by acoustic similarity reflect genetic groupings, using AFLP data to estimate genetic relationships. Second, we tested whether genetic boundaries are maintained by female preferences for conspecific song characteristics. Phonotaxis trials were used to determine the extent of female preferences for conspecific male song. Results generally support both hypotheses, but suggest the presence of porous species boundaries. PMID- 12555787 TI - Domesticated birds as a model for the genetics of speciation by sexual selection. AB - In theory, even populations occupying identical environments can diverge in sexually selected traits, as a consequence of different mutational input. I evaluate the potential of this process by comparing the genetics of breeds of domesticated birds to what is known about the genetics of differences among species. Within domesticated species there is a strong correlation of time since domestication with the number of breeds. Descendants of the rock dove, Columba livia (the oldest domesticate) show differences in courtship, vocalizations, body shape, feather ornaments (crests and tails) and colors and color patterns. When nine other domesticated species are included there is a striking hierarchy, with more recent domesticates having a nested subset of these traits: the youngest domesticated species have breeds distinguished only by color. This suggests that selection of new, visible, mutations is driving the process of breed diversification, with mutations that appeal to the breeder happening the most frequently in color. In crosses among related species, color, feather ornaments and many vocalizations and displays show both intermediate dominance and pure dominance. Although the number of loci affecting each of these traits is typically unknown, limited evidence of the genetics of species' differences suggests that some differences are due to the substitution of single genes of major effect. While neither the genetics of breeds nor the genetics of species provide a perfect model for the genetics of speciation, similarities between the two are sufficiently striking to infer that major, visible, mutations can provide the impetus underlying new directions of sexual selection. PMID- 12555788 TI - Patterns of inheritance of mating signals in interspecific hybrids between sailfin and shortfin mollies (Poeciliidae: Poecilia: Mollienesia). AB - Differences in male morphology and mating behaviors are thought to confer species sexual isolation between sailfin and shortfin species of mollies. This study used interspecific crosses between the sailfin molly, P. latipinna, and the shortfin molly, P. mexicana, to investigate patterns of inheritance of morphological traits and behavioral rates of three mating behaviors in F1 hybrid males. The two parental species showed clear species differences with respect to the length of the dorsal fin and dorsal fin ray number. First generation hybrid males were intermediate between the two parental species for dorsal fin length and fin ray number, suggesting autosomal control of this trait with little effect of dominance by genes from either parental species. Parental species showed clear species differences in their rates of courtship displays. Unlike the pattern for dorsal fin morphology, F1 hybrid males showed a clear distinction in display rates with respect to the direction of the interspecific cross. Male hybrids whose sires were P. latipinna had courtship display rates that were up to three times higher than the rates of displays performed by hybrid males whose fathers were P. mexicana. The distribution of phenotypes between the parental species and that of hybrid males sired by that parental species was nearly identical. Such a pattern suggests the influence of Y-linked genes on the inheritance of courtship display rates in mollies. PMID- 12555789 TI - Variation in female mate choice within guppy populations: population divergence, multiple ornaments and the maintenance of polymorphism. AB - The evolutionary significance of variation in mate choice behaviour is currently a subject of some debate and considerable empirical study. Here, I review recent work on variation within and among guppy (Poecilia reticulata) populations in female mate choice and mating preferences. Empirical results demonstrate that there is substantial variation within and among populations in female responsiveness and choosiness, and much of this variation is genetic. Evidence for variation in preference functions also exists, but this appears to be more equivocal and the relative importance of genetic variation is less clear cut. In the second half of this review I discuss the potential significance of this variation to three important evolutionary issues: the presence of multiple male ornaments, the maintenance of polymorphism and divergence in mate recognition among populations. Studies of genetic variation in mate choice within populations indicate that females have complex, multivariate preferences that are able to evolve independently to some extent. These findings suggest that the presence of multiple male ornaments may be due to multiple female mating preferences. The extreme polymorphism in male guppy colour patterns demands explanation, yet no single satisfactory explanation has yet emerged. I review several old ideas and a few new ones in order to identify the most promising potential explanations for future empirical testing. Among these are negative frequency dependent selection, environmental heterogeneity coupled with gene flow, and genetic constraints. Last, I review the relative extent of within and among-population variation in mate choice and mating preferences in order to assess why guppies have not speciated despite a history of isolation and divergence. I argue that variation within guppy populations in mate choice and enhanced mating success of new immigrants to a pool are major impediments to population divergence of the magnitude that would be required for speciation to occur. PMID- 12555790 TI - Sexual isolation and speciation in bacteria. AB - Like organisms from all other walks of life, bacteria are capable of sexual recombination. However, unlike most plants and animals, bacteria recombine only rarely, and when they do they are extremely promiscuous in their choice of sexual partners. There may be no absolute constraints on the evolutionary distances that can be traversed through recombination in the bacterial world, but interspecies recombination is reduced by a variety of factors, including ecological isolation, behavioral isolation, obstacles to DNA entry, restriction endonuclease activity, resistance to integration of divergent DNA sequences, reversal of recombination by mismatch repair, and functional incompatibility of recombined segments. Typically, individual bacterial species are genetically variable for most of these factors. Therefore, natural selection can modulate levels of sexual isolation, to increase the transfer of genes useful to the recipient while minimizing the transfer of harmful genes. Interspecies recombination is optimized when recombination involves short segments that are just long enough to transfer an adaptation, without co-transferring potentially harmful DNA flanking the adaptation. Natural selection has apparently acted to reduce sexual isolation between bacterial species. Evolution of sexual isolation is not a milestone toward speciation in bacteria, since bacterial recombination is too rare to oppose adaptive divergence between incipient species. Ironically, recombination between incipient bacterial species may actually foster the speciation process, by prohibiting one incipient species from out-competing the other to extinction. Interspecific recombination may also foster speciation by introducing novel gene loci from divergent species, allowing invasion of new niches. PMID- 12555791 TI - Mutation and premating isolation. AB - While premating isolation might be traceable to different genetic mechanisms in different species, evidence supports the idea that as few as one or two genes may often be sufficient to initiate isolation. Thus, new mutation can theoretically play a key role in the process. But it has long been thought that a new isolation mutation would fail, because there would be no other individuals for the isolation-mutation-carrier to mate with. We now realize that premeiotic mutations are very common and will yield a cluster of progeny carrying the same new mutant allele. In this paper, we discuss the evidence for genetically simple premating isolation barriers and the role that clusters of an isolation mutation may play in initiating allopatric, and even sympatric, species divisions. PMID- 12555792 TI - Female choice in Drosophila: evidence from Hawaii and implications for evolutionary biology. AB - Details of female choice of mate in Drosophila silvestris of Hawaii strikingly parallels epigamic behavioral systems in many other animals and may be common in other species of Drosophilidae. Females respond selectively to male circling, wing displays, songs and tactile stimulation with foreleg cilia, a quantitative character that is highly variable in some populations. I hypothesize that the female can exert choice based on these cues from individual males that differ genetically by quantitative trait loci. Laboratory tests show that one third of courting males are repeatedly rejected in favor of a minority of 'alpha' males. This result imposes non-random mating at the local population level. Past multiple-choice lab tests, widely used to measure 'isolation' between pairs of populations or species of Drosophila may be flawed, since random mating has been assumed in the interpretation of results. Pre-mating sexual selection is clearly a powerful intrapopulation force in population biology. This view creates difficulties for discerning any proposed simultaneous interpopulation selective events in the presence of strong female choice. The long-held theory assuming that there is significant selection for pre-mating isolation between groups is questionable. PMID- 12555793 TI - Understanding the association between socioeconomic status and physical health: do negative emotions play a role? AB - In this article, the authors evaluate the possible roles of negative emotions and cognitions in the association between socioeconomic status (SES) and physical health, focusing on the outcomes of cardiovascular diseases and all-cause mortality. After reviewing the limited direct evidence, the authors examine indirect evidence showing that (a) SES relates to the targeted health outcomes, (b) SES relates to negative emotions and cognitions, and (c) negative emotions and cognitions relate to the targeted health outcomes. The authors present a general framework for understanding the roles of cognitive-emotional factors, suggesting that low-SES environments are stressful and reduce individuals' reserve capacity to manage stress, thereby increasing vulnerability to negative emotions and cognitions. The article concludes with suggestions for future research to better evaluate the proposed model. PMID- 12555794 TI - Predictors of posttraumatic stress disorder and symptoms in adults: a meta analysis. AB - A review of 2,647 studies of posttraumatic stress disorder (PTSD) yielded 476 potential candidates for a meta-analysis of predictors of PTSD or of its symptoms. From these, 68 studies met criteria for inclusion in a meta-analysis of 7 predictors: (a) prior trauma, (b) prior psychological adjustment, (c) family history of psychopathology, (d) perceived life threat during the trauma, (e) posttrauma social support, (f) peritraumatic emotional responses, and (g) peritraumatic dissociation. All yielded significant effect sizes, with family history, prior trauma, and prior adjustment the smallest (weighted r = .17) and peritraumatic dissociation the largest (weighted r = .35). The results suggest that peritraumatic psychological processes, not prior characteristics, are the strongest predictors of PTSD. PMID- 12555795 TI - Cues to deception. AB - Do people behave differently when they are lying compared with when they are telling the truth? The combined results of 1,338 estimates of 158 cues to deception are reported. Results show that in some ways, liars are less forthcoming than truth tellers, and they tell less compelling tales. They also make a more negative impression and are more tense. Their stories include fewer ordinary imperfections and unusual contents. However, many behaviors showed no discernible links, or only weak links, to deceit. Cues to deception were more pronounced when people were motivated to succeed, especially when the motivations were identity relevant rather than monetary or material. Cues to deception were also stronger when lies were about transgressions. PMID- 12555796 TI - Forewarned and forearmed? Two meta-analytic syntheses of forewarnings of influence appeals. AB - Two research syntheses evaluate the effects on attitudes of forewarning of an influence appeal. In general, warnings appeared to threaten people's attitudes or their self-images, and warning impact depended on which aspect of the self was threatened. When the topic of the appeal was involving and concerned immediate outcomes or when the appeal was actually delivered, recipients appeared to focus on the potential threat to their attitudes, and they responded defensively by cognitively bolstering their own views and resisting the appeal. However, warnings of appeals on less involving topics generated agreement before the appeal was delivered, presumably because these warnings alerted people to the self-image threat of being gullible and preemptive agreement reduced this threat. PMID- 12555797 TI - The psychology of doing nothing: forms of decision avoidance result from reason and emotion. AB - Several independent lines of research bear on the question of why individuals avoid decisions by postponing them, failing to act, or accepting the status quo. This review relates findings across several different disciplines and uncovers 4 decision avoidance effects that offer insight into this common but troubling behavior: choice deferral, status quo bias, omission bias, and inaction inertia. These findings are related by common antecedents and consequences in a rational emotional model of the factors that predispose humans to do nothing. Prominent components of the model include cost-benefit calculations, anticipated regret, and selection difficulty. Other factors affecting decision avoidance through these key components, such as anticipatory negative emotions, decision strategies, counterfactual thinking, and preference uncertainty, are also discussed. PMID- 12555798 TI - Knowledge, attitudes, and behavior of women ages 65 and older on mammography screening and Medicare: results of a national survey. AB - BACKGROUND: Compared to younger women, women 65+ will experience the greatest increase in new breast cancer cases. In 1991, Medicare began offering partial reimbursement for screening mammography every 2 years. METHODS: In 1999, the National Cancer Institute (NCI) conducted a telephone survey on breast cancer, mammography, and Medicare reimbursement with a sample of households containing women ages 65+ using random-digit-dialing. Results were weighted to provide nationally representative estimates of U.S. women 65+. NCI compared 1999 results with similar data from a 1992 AARP survey. RESULTS: Of the 814 women surveyed, 88% had had at least one mammogram in their lifetime; within this group, 80% had received their most recent mammogram 2 years ago or less. Only 57%, however, knew about recommendations to have a mammogram every 1-2 years. Approximately one third indicated that they were not as concerned about getting breast cancer as when they were younger, and/or that women without risk factors could be less vigilant about mammograms. More than 75% were aware of Medicare coverage, but only 58% had used Medicare to help pay for their last mammogram. Minority women were almost twice as likely to be unaware of Medicare coverage. RECOMMENDATIONS: (1) Highlight that breast cancer risk increases with age (and does not decline in the absence of risk factors) and communicate the correct frequency for having mammograms; (2) expand primary care physicians' roles in promoting mammography screening for women 65+; and (3) provide Medicare coverage information to older women, particularly those not taking advantage of this benefit. PMID- 12555799 TI - Women's knowledge and experience of atypical Pap results in a high risk community. AB - We sought to describe the knowledge of, understanding of, and follow-up after atypical cervical cytology among low-income, urban women. Of 554 women who had an ASCUS or atypical Pap result in 1996,215 (39%) completed a telephone survey. Many (19%) had poor follow-up. Thirty percent of women denied ever being told of abnormal cytology; Spanish-speakers were more likely to be unaware (43.2% vs. 22.7%, p < .01) of abnormal cytology. Cancer fear was quite high, as were misconceptions regarding cause. Many women were unsatisfied with the information they received regarding their cytology. PMID- 12555800 TI - Effects of dumbbell and elastic band training on physical function in older inner city African-American women. AB - Providing inner-city minorities with inexpensive exercise programs may be a means of reducing frailty and maintaining independent lifestyles in this population. To determine the efficacy of such a program, 22 African-American women aged 62-94 years were recruited from a senior center in the Wichita urban community. Sixteen women (age = 74.8 +/- 8.8 yr) participated in exercise training (3 days per week for 4 weeks) and 6 women (age = 74.7 +/- 4.5 yr) served as controls. Training consisted of chair-based exercises using elastic resistance bands (upper and lower body) and dumbbells (upper body). Elastic band training improved (p < 0.05) lower body strength (30-second chair stand, time for 5 stands) by approximately 20% and the combination of bands and dumbbell training improved upper body strength (30-second dumbbell curl) by 24%. Grip strength improved by 5%. Up-&-Go performance improved by 10% although balance and mobility exercises were not a fundamental element of the training program. Upper and lower body flexibility did not change. Changes were not observed for any variable in the control group. This study suggests that exercising with equipment that costs only a few dollars per participant improves upper and lower body fitness in older African-American women who live in an urban setting. PMID- 12555801 TI - Life event timing and the emotional consequences of surgical menopause for Asian Pacific women in Guam. AB - Using Behavior Risk Factor Survey data (N = 398), this study tests the age-norms hypothesis by examining the emotional consequences of the timing of surgical menopause for women in Guam's Asian-Pacific community. The results of the study are supportive of the hypothesis that off-scheduled life events result in considerable emotional trauma, and show that women in Guam who are 44 years of age or younger are significantly more likely than older women to suffer psychological distress and unhappiness when they report experiencing surgical menopause. We discuss the potential implications of these findings for theory and future research. PMID- 12555802 TI - Attitude toward menopause among married middle-aged adults. AB - One-hundred and sixty-nine-married couples participated in a study designed to examine the relationship of menopausal attitude to the frequency of menopausal symptoms at midlife. Participants completed the Bowles' (1986a) Menopause Attitude Scale and a modified version of the Menopausal Symptoms Checklist (Neugarten & Kraines, 1965). Wives expressed a more positive attitude toward menopause than their husbands, and wives reported experiencing more menopausal symptoms than their husbands perceived them as having. Post-menopausal women expressed a more positive attitude toward menopause than peri-menopausal women or women who had experienced surgical menopause. For both men and women, a positive attitude toward menopause was associated with women who reported fewer menopausal symptoms. A preliminary Structural Equation Model is presented as an effort to further stimulate the development of a theoretical model of attitude toward menopause among middle-aged married couples. PMID- 12555803 TI - Relationship power, sexual decision making and condom use among women at risk for HIV/STDS. AB - This study examines the associations among relationship power, sexual decision making dominance, and condom use within a sample of women at risk of HIV/STDs. Data from face-to-face interviews with 112 women were analyzed to (a) describe who women perceive as more powerful and who makes sexual decisions within their heterosexual relationships, (b) explore the association between relationship power and sexual decision-making dominance, and (c) examine the relationship of power and decision making regarding condom use to condom use behavior. Women were recruited from clinics and community locations in Atlanta, Los Angeles, Oklahoma City and Portland, OR. Participants were 18-25 years of age and were primarily Hispanic and African American. Over half (58.2%) reported that they share power with their partner, 25.5% said they have more power, and 16.4% reported that their partner has more power in their relationship. For the five domains of sexual decision-making examined, over half (50.5%-75.7%) of the women reported that they and their partners make decisions together. A higher percentage of women who perceived that they have more power or share power, as compared to those who perceived that their partners have more power, reported that "I/We" make decisions about birth control use, condom use, whether to have sex, and type of sexual activity. Relationship power was not associated with condom use. Condom use was, however, significantly higher among women who reported that they make decisions about using condoms alone or with their partner as compared to those who reported that their partner makes those decisions. PMID- 12555804 TI - Legitimacy aspects of social network relations in a group of women with undefined musculoskeletal disorder. AB - This study was conducted to investigate legitimacy aspects of social network relations in a group of women with undefined musculoskeletal disorder. The aim was to study, from the viewpoint of the patient, the extent to which the symptoms of musculoskeletal disorder have legitimacy within the social network and how this legitimacy affects the patient's social network relations. A structural equation model was used to find the model best fitting the data arising between social support, legitimacy aspects and negative outcomes in the social network relations. The results can be summarised as indicating that illness legitimacy does affect availability of social relations and social support from friends, relatives and family. Of particular importance to this process is the patient's own perception of the legitimacy of her disorder. These results also confirmed previous findings indicating that greater attention should be paid to the impact of negative aspects of social support and its affect on patient health. PMID- 12555805 TI - From pragmatism to politics: a qualitative study of abortion providers. AB - Twenty-eight years after the United State Supreme Court issued its landmark Roe v. Wade, the struggle continues to ensure that all women have the full range of reproductive choices, including abortion. While the struggle can be addressed through its political, religious, and medical dimensions, it also can be examined through the perspectives of those who actually provide abortions. This paper examines the perspectives of physician abortion providers to understand more fully their motivations, the quality of their personal and professional lives, their views on the future of abortion services, and their recommendations for undergraduate and residency medical education. Such questions are often best answered through qualitative inquiry, particularly when the subject at hand has had little interpretive scrutiny, lacks theoretical understandings, and remains in general an under-investigated phenomenon. Because abortion providers and the work they do fit those criteria, a qualitative study of physician providers in Ohio was undertaken. This paper is divided into the following sections: a literature review of abortion services in the United States, methods, interview data and discussion, and last, recommendations and conclusions. PMID- 12555806 TI - Explanatory variance in bulimia nervosa. AB - In order to prevent disease, one must understand the factors that contribute to the variance of the illness. In an effort to better understand factors that contribute to bulimia nervosa (BN), a study was designed to investigate explanatory factors in BN. Of particular interest to the researchers in the study was the effect of the Hispanic subculture on the variance of BN. Female participants (n = 372) and male participants (n = 109) from the ages of 17-49 agreed to serve in the study. A self-report scale designed to assess the symptoms of BN was the criterion variable in the study. Predictor variables included in the regression model were drive for thinness, body mass index (BMI), age, and ethnicity (Hispanic or Anglo). Separate analyses were run for males and females. Based on the results of the analysis it seems that the desire to be thinner and an individual's BMI contribute to the variance in BN for females, but not for males. There was no difference in the attitudes associated with BN between Hispanic (n = 27) and Anglo (n = 321) females. PMID- 12555807 TI - Urea and KCl have differential effects on enzyme activities in liver and muscle of estivating versus nonestivating species. AB - The effects of 300 mM urea or 300 mM KCl on the maximal activities of 25 enzymes of intermediary metabolism were assessed in extracts of liver and muscle from spadefoot toads (Scaphiopus couchii), leopard frogs (Rana pipiens), and rats to assess their sensitivity to these osmolytes. During estivation, toads can lose 50% of total body water, and urea, which is known for its action as a protein denaturant, accumulates to 200-300 mM. The data show that the maximal activities of toad liver enzymes were not affected when assayed in the presence of 300 mM urea in vitro whereas urea inhibited the activities of seven enzymes in frog and 11 enzymes in rat liver. High KCl affected 12 or 13 enzymes in liver of each species causing inhibition in eight or nine cases each, and for frog and rat enzymes, inhibition was frequently more pronounced than for urea. Both urea and KCl affected enzyme activities in muscle extracts of all three species, but whereas their effects were largely negative for frog and rat enzymes, the enzymes affected by urea or KCl in toad muscle were primarily activated by these osmolytes (six out of nine enzymes affected by urea and eight of 15 enzymes affected by KCl). Urea, KCl, and polyethylene glycol (a protein crowding agent) also had species-specific effects on the dissociation constant (Ka) for cAMP of protein kinase A. The data suggest that the accumulation of urea by water stressed anurans not only contributes to minimizing cell volume reduction, but by doing so also limits the increase in intracellular ionic strength that occurs and thereby helps to minimize the potential inhibitory effects of high salts on metabolic enzymes. PMID- 12555808 TI - Increased reactive oxygen species production with antisense oligonucleotides directed against uncoupling protein 2 in murine endothelial cells. AB - Uncoupling protein 2 (UCP-2) belongs to the mitochondrial anion carrier family. It is ubiquitously expressed but is most abdundant in the reticuloendothelial system. In addition to uncoupling function, UCP-2 modulates the production of reactive oxygen species (ROS) by isolated mitochondria. Using an antisense oligonucleotide strategy, we investigated whether a defect in UCP-2 expression modulates ROS in intact endothelial cells. Murine endothelial cells (CRL 2181) pretreated by antisense oligonucleotides directed against UCP-2 mRNA exhibited a significant and specific increase in membrane potential and intracellular ROS level compared with control scrambled or anti-UCP-1 and -UCP-3 antisense oligonucleotides. These specific changes induced by UCP-2 antisense oligonucleotides were correlated with a rise in extracellular superoxide anion production and oxidative stress assessed by thiobarbituric acid reactive substance values. Taken together, these data suggest a role for UCP-2 in control of ROS production and subsequent oxidation of surrounding compounds mediating oxidative stress of endothelial cells. These data also support the notion that manipulations of UCP-2 at the genetic level could control ROS metabolism at the cellular level. PMID- 12555809 TI - High-mobility group protein 2 may be involved in the locus control region regulation of the beta-globin gene cluster. AB - Expression regulation of the beta-globin gene cluster is a result of synergistic interactions between cis-elements and trans-acting factors. Previous studies usually concentrated on the core sequence of each hypersensitive site in the locus control region of the beta-globin gene cluster. But more and more evidence illustrates that the flanking regions are indispensable also. Using electrophoretic mobility shift assay and solid-phase DNase I footprinting methods, we identified a small nuclear protein from K562 cells that binds specifically to the first AT-rich region flanking the hypersensitive site 2 core sequence of the human beta-globin gene locus control region. N-terminal sequencing of the enriched protein proved that it is a member of the high mobility group protein 2 family. This indicates that the AT-rich region in human hypersensitive site 2 may take part in the regulation of the beta-globin gene cluster by facilitating DNA bending, which is a prerequisite for the looping mechanism in this region. PMID- 12555810 TI - Regulatory properties of p105: a novel PKC isoenzyme in mantle tissue from marine mussels. AB - Previous results suggested operative similarities between Apl II from nervous cells of Aplysia californica, epsilonPKC from brain of vertebrates, and p105 from mantle tissue of Mytilus galloprovincialis Lmk., all of them belonging to the nPKC family. The optimal substrate for Apl II and p105 from mussel is protamine sulfate. In contrast, Ca2+ inhibits p105 but does not affect Apl II. As occurs in other epsilonPKC, p105 is autophosphorylable; however, in Apl II, no P-Tyr residues are detected in the most phosphorylated form. The presence of p105 in all the tissues of M. galloprovincialis studied, proves the important, yet unknown, physiological role that this enzyme must play. PMID- 12555811 TI - Secretion of foreign proteins mediated by chicken lysozyme gene regulatory sequences. AB - Exploitation of the insulating properties of the complete chicken lysozyme gene domain may facilitate the production of transgenic chicken bioreactors with the capacity to deposit valuable proteins in the egg white. Chimeric genes consisting of the chicken lysozyme gene regulatory sequences and sequences encoding foreign proteins could be inserted randomly into the chicken genome and retain appropriate expression levels. The research reported here established that chicken lysozyme gene regulatory sequences can be used to direct the production and secretion of green fluorescent protein (used as a reporter protein) in transiently transfected chicken blastodermal cells. Attempts to verify these findings in transgenic hens are currently in progress. To provide a rapid means of generating constructs encoding other foreign proteins under the control of lysozyme gene regulatory sequences that can facilitate the secretion of heterologous proteins in vivo, a generic lysozyme gene regulatory scaffold was created using a poxvirus-mediated gene targeting system. PMID- 12555813 TI - The oligomeric state, complex formation, and chaperoning activity of hsp70 and hsp80 of Neurospora crassa. AB - Molecular chaperones perform vital cellular functions under normal growth conditions and protect cells against stress-induced damage. The stress proteins Hsp70 and Hsp80 of Neurospora crassa were extracted from heat-shocked mycelium, purified to near homogeneity, and examined with respect to their oligomeric state, complex formation, and chaperoning properties. Their oligomeric state was assessed by dynamic light-scattering measurements, and both Hsp70 and Hsp80 were observed to form a range of soluble, high-molecular-mass protein aggregates. Direct interaction between Hsp70 and Hsp80 was studied by partial tryptic digestion and surface plasmon resonance (SPR). Hsp70 was immobilized on the sensor chip surface, and the binding of Hsp80 in solution was followed in real time. Proteolytic digestion revealed that Hsp70-Hsp80 complex formation results in conformational changes in both proteins. The data from SPR studies yielded an equilibrium dissociation constant, KD, of 8.5 x 10(-9) M. The chaperoning ability of Hsp70, Hsp80, and Hsp70-Hsp80 was monitored in vitro by the protection of citrate synthase from thermal aggregation. The binding of nucleotides modulates the oligomeric state, chaperoning function, and hetero-oligomeric complex formation of Hsp70 and Hsp80. PMID- 12555812 TI - Cellular resistance to bleomycin in Saccharomyces cerevisiae is not affected by changes in bleomycin hydrolase levels. AB - Bleomycin is a glycopeptide drug that exerts potent genotoxic potential and is highly effective in the treatment of certain cancers when used in combination therapy. Unfortunately, however, tumors often develop resistance against bleomycin, and the mechanism of this resistance remains unclear. It has been postulated that bleomycin hydrolase, a protease encoded by the BLH1 gene in humans, may account for tumor resistance to bleomycin. In support of such a notion, earlier studies showed that exogenous expression of yeast Blh1 in human cells can enhance resistance to bleomycin. Here we show that (i) yeast blh1delta mutants are not sensitive to bleomycin, (ii) bleomycin-hypersensitive yeast mutants were no more sensitive to this agent upon deletion of the BLH1/LAP3/GAL6 gene, and (iii) overproduction of Blhl in either the parent or bleomycin hypersensitive mutants did not confer additional resistance to these strains. Therefore, yeast Blh1 apparently has no direct role in protecting this organism from the lethal effects of bleomycin, even though the enzyme can degrade the drug in vitro. Clearly, additional studies are required to establish the actual biological role of Blh1 in yeast. PMID- 12555815 TI - Science and nursing education. PMID- 12555814 TI - Detection of multiple splice variants of the nuclear protein phosphatase 1 regulator sds22 in rat liver nuclei. AB - Antipeptide antibodies generated against the N terminus of the protein phosphatase 1 (PP1) binding protein sds22 detected at least four forms of the protein in a rat liver nuclear extract. Four of these immunoreactive bands likely correspond to four predicted forms of sds22 that are generated by alternative splicing. These four proteins are expressed at different levels and appear to be localized exclusively in the nucleus, and two of these proteins copurify with PPI on the protein phosphatase affinity matrix microcystin-Sepharose. Two higher molecular mass nuclear proteins that are immunoreactive with the sds22 antibodies also copurify on microcystin-Sepharose and may be novel forms of sds22 expressed in mammalian cells. PMID- 12555816 TI - My heart is naked. PMID- 12555817 TI - Integrating mental illness prevention into community-based undergraduate education. AB - Including preventive models of health care in undergraduate education is essential as the nursing profession moves increasingly to community-based care. Traditionally, mental health curricula have focused primarily on psychopathology. This article presents current research in the attachment and temperament literature, which provides sound evidence for the need to synthesize prevention of mental illness concepts into undergraduate education. A curriculum model that integrates concepts from psychiatry and public health to prepare nursing students to promote mental health is described. PMID- 12555818 TI - Critical thinking: perceptions of newly graduated female baccalaureate nurses. AB - Nursing education strives to develop critical thinking through emphasis on process, inquiry, and reasoning. Although North American nursing education programs have recommended integrating the critical thinking construct into the conceptual framework that supports undergraduate nursing programs, critical thinking is still ambiguously and inconsistently applied within the profession. The research described in this article explored the development of thinking in five newly graduated baccalaureate RNs by accompanying them on the conceptual, theoretical, and practical evolutionary journey of their first 6 months in nursing practice and explicating their knowledge development over time, offering insights into the role of undergraduate education in teaching, and fostering critical thinking as an approach to nursing practice. PMID- 12555819 TI - Complementary and alternative medicine: core competencies for family nurse practitioners. AB - Recent surveys demonstrate substantial use of complementary and alternative medicine. This growth presents a challenge for family nurse practitioner programs that already have dense curricula due to the explosion of biomedical knowledge. Significant barriers exist to incorporating complementary and alternative medicine into educational programs, such as differing philosophies about health and lack of research on many complementary and alternative medicine therapies. This study used a mailed questionnaire in which family nurse practitioner program directors described the current status of complementary and alternative medicine teaching in family nurse practitioner programs and identified core competencies for family nurse practitioners. Most programs reported integrating some content on complementary and alternative medicine into advanced practice courses, such as health promotion. Competencies related to interviewing, critical thinking, evidence-based medicine, knowledge of laws, ethics, and spiritual and cultural beliefs were rated the highest. The findings are useful in helping family nurse practitioner faculty incorporate complementary and alternative medicine content into their curricula with the goal of preparing family nurse practitioners for safe practice. PMID- 12555820 TI - Guidelines for developing and implementing humor in nursing classrooms. AB - The three teachers in this study selected many types of humor to exemplify their points. To avoid predictability, they varied their humor presentation styles and frequency by using simple cartoons projected on a screen, jokes and stories, and songs and dance routines. All three teachers engaged in a continuous search for new material to further expand their repertoire. Careful not to overuse humor, they never lost sight of the learning objectives, but they added enough humor to sustain students' attention. The teachers gradually increased their use of classroom humor as they felt more comfortable with the content and setting. All of the students' and teachers' comments corroborate the conclusions reached by Bryant and Zillmann 1989), "It would appear that success in teaching with humor...depends on employing the right type of humor, under the proper conditions, at the right time, and with properly motivated and receptive students' (p. 74). Not all types of humor are appropriate for all teachers. However, for teachers who carefully invest time and effort into learning to teach with humor, the reward can be a successful and enjoyable experience for all involved. PMID- 12555821 TI - Posters: an assessment strategy to foster learning in nursing education. PMID- 12555822 TI - Clinical nursing instructors' use of handheld computers for student recordkeeping and evaluation. AB - I have found the Palm IIIc to be useful in tracking undergraduate student progress in the clinical area. Entrepreneurial thinkers continue to develop new ideas in software, so the possibilities for using these devices will only grow. Future possibilities for handheld computers include built-in Internet connections or providing students with devices to record and analyze their experiences or complete assignments. Many nurses and physicians who have learned to use handheld computers are sure they never want to go back to using paper and pen (Labkoff & O'Mahony, 1997; Wilson & Fulmer, 1998). I will continue to depend on and find new uses for this pocket-sized, yet powerful, device. PMID- 12555823 TI - Evaluation of faculty transition into a community-based curriculum. PMID- 12555824 TI - Driving forces maps: case studies of a tool for implementing change strategies. AB - In this article, driving forces mapping, a technique derived from business strategy, is applied to five different client situations: an AIDS support organization, a men's bereavement group, an individual managing grief after abortion following diagnosis of a fetal anomaly, an individual dealing with an HIV diagnosis, and a couple who wanted to increase marital satisfaction. The technique was found to be helpful in goal definition, contracting and developing mutuality, and it was less helpful in introspective work. The article describes the work in ways that allow the reader to understand the process and apply the technique. PMID- 12555825 TI - The impact of a computer network on pediatric pain and anxiety: a randomized controlled clinical trial. AB - This was the third randomized controlled clinical trial (RCCT) of Starbright World (SBW)--a private, interactive computer network for hospitalized children. The present randomized controlled clinical trial (RCCT) consisted of a series of 39 replicated single system designs (SSDs) that compared children's self-reports of pain intensity, pain aversiveness and anxiety in the SBW condition with those in a stan- dard clinical practice condition. Findings from the SSDs were aggregated using meta-analysis. Although children experienced slightly less pain intensity, pain aversiveness, and anxiety in the SBW condition, these results did not achieve statistical significance. The relationship of these findings to the two prior studies of the effectiveness of SBW is discussed. PMID- 12555826 TI - Using DAVE as a model to go beyond treatment compliance with persons receiving renal replacement therapy. AB - DAVE is an acronym for Dialogue, Accept, Validate, and Enact conceptualized for the purpose of helping nephrology social workers assist persons with End-Stage Renal Disease take an active role in making treatment decisions, especially the decision to comply with a treatment regimen. As a model, DAVE is a summation of contemporary social work practice including views on empowerment, the Strengths Perspective, and the Problem-Solving Process. DAVE attunes the nephrology social worker and the dialysis recipient to an attitude of mutual trust, respect, and collaboration essential to helping dialysis recipients make important treatment decisions. PMID- 12555827 TI - Providing therapy when managed care organizations deny reimbursement: are private practitioners and their clients taking the fall? AB - Sampled private practitioners from the Register of Clinical Social Workers, American Psychological and Psychiatric Associations were used to evaluate what percentage of these practitioners' caseloads are made up of clients who have been denied reimbursement by managed care organizations for ongoing therapy. The results indicate that 39% of the respondents' caseloads are of managed care insured clients, of whom nearly one-third have been denied reimbursement for ongoing therapy, and of whom 42% later reenter therapy with an acute exacerbation of symptoms. The results have potential ethical and legal complications for private practitioners treating clients insured by managed care organizations. PMID- 12555828 TI - Separating the apples and oranges in the fruit cocktail: the mixed results of psychosocial interventions on cancer survival. AB - In past reviews of the literature on psychosocial interventions for cancer patients, different reviewers have reached disparate conclusions about the ability of a psychosocial intervention to have an impact on cancer survival. This article highlights some of the basis for differences of opinion, including the range of psychosocial interventions provided to patients and the challenges inherent in conducting research studies of this type. Social workers who work with cancer patients at vulnerable times need to understand what the current evidence shows can be gained from participation in a psychosocial intervention. Several key questions have been identified that can provide the basis for future rigorous studies. The role of the social worker in participating and shaping research on the relationship of psychosocial intervention and cancer survival is also discussed. PMID- 12555829 TI - Radiologic case study. Radial shaft fracture, anterior interosseous nerve injury, and the presence of a foreign body within the soft tissues of the proximal forearm. PMID- 12555830 TI - Cemented stem failure requires extended trochanteric osteotomy. PMID- 12555831 TI - The extended femoral slot: an alternative method for cemented stem failure. PMID- 12555832 TI - Subvastus approach for total knee arthroplasty. AB - The indications for the subvastus approach in total knee arthroplasty are the same for a medial parapatellar approach in the appropriately selected patient. The subvastus approach does not violate the quadriceps mechanism, reduces the need for lateral retinacular release, preserves the patella blood supply, and, with our modifications, can be used in most cases. PMID- 12555833 TI - Alumina ceramic bearings for total hip arthroplasty. AB - This article presents an experience with alumina ceramic bearings involving the use of improved ceramic materials as well as new design considerations. The alumina-alumina ceramic hard bearing is a safe option for the younger, more active patient. PMID- 12555834 TI - The economic impact of medical complications in geriatric patients with hip fracture. AB - The records of 510 elderly patients with hip fractures admitted to our institution between January 1995 and December 2000 were retrospectively reviewed to determine the incidence and type of developed medical complications and their economic implications. Of those 510 patients, 217 (43%) developed at least 1 medical complication, most frequently electrolyte imbalance (11%), urinary tract infection (10%), respiratory failure (10%), and delirium (9%). Patients who developed medical complications had significantly longer mean hospital stays (10 days) and higher mean hospital costs ($16,203) than patients without such complications (5 days and $10,284, respectively) (P<.001). PMID- 12555835 TI - Outcomes of hip fracture surgery in patients aged > or = 90 years. AB - The outcomes of 60 patients aged > or = 90 years with hip fractures who underwent surgery between 1995 and 1998 were reviewed. Average follow-up was 21 months (range: 4-47 months). All patients experienced pain relief within 2 months postoperatively. Thirty-four of 60 patients failed to attain their preinjury ambulatory status. The ability to walk 2 months postoperatively was observed more frequently in patients without severe dementia. Patients who were unable to walk postoperatively had a greater chance of dying than those who regained the ability to walk. No surgery related deaths occurred. Six months postoperatively, 2 (3%) patients had died; 1 year postoperatively, 9 (15%) had died. Therefore, patients aged > or = 90 years benefit from surgical treatment of hip fractures. PMID- 12555836 TI - The effectiveness of a pain wrap compared to a standard dressing on the reduction of postoperative morbidity following routine knee arthroscopy: a prospective randomized single-blind study. AB - A pain wrap dressing in patients undergoing routine knee arthroscopy was evaluated to determine its ability to decrease postoperative pain and swelling. Bilateral knee examinations were performed pre- and postoperatively. Patients recorded narcotic usage and pain levels for 10 postoperative days. Twenty-four patients completed the study. The pain wrap group had lower pain ratings (2.2 versus 4.6 [P=.03]) and demonstrated lower skin temperatures (1.1 degrees F versus 3.9 degrees F [P=.02]). Less postoperative swelling was noted in the treatment patients, whereas postoperative arc of motion and narcotic usage was similar in both groups. PMID- 12555837 TI - Magnetic resonance imaging of tuberculous spondylitis. AB - The clinical and imaging findings of patients with a confirmed diagnosis of tuberculous spondylitis were retrospectively analyzed to assess the diagnostic value of magnetic resonance imaging (MRI) and determine the different patterns of spine involvement. Fifty-three patients with plain radiographs and MRI of the entire spinal column were included in the study. Sagittal T1- and T2-weighted MRIs of the entire spine and axial T1- and T2-weighted MRIs at the levels of interest were retrospectively evaluated. Plain radiographic correlation was obtained in all patients. The lower thoracic and thoracolumbar spine was the most commonly involved region. Magnetic resonance imaging is effective in the early diagnosis of tuberculous spondylitis. It also detects lesions, which may not be apparent on plain radiographs. PMID- 12555838 TI - Pregnancy outcomes following total hip arthroplasty: a preliminary study and review of the literature. AB - Pregnancy outcomes among patients who underwent total hip arthroplasty (THA) during their reproductive years were retrospectively evaluated. Twenty-one patients reported pregnancies after THA and 20 had live births. No prosthesis related problems were reported. This is the first study that provides a patient based assessment of pregnancy outcomes and delivery in women who underwent THA. The preliminary data suggest THA had no adverse effect on subsequent childbearing. PMID- 12555839 TI - Osteoinduction by a collagen mineral composite combined with isologous bone marrow in a subcutaneous rat model. AB - In a subcutaneous implant rat model, Collagraft alone (n=8), Collagraft plus isologous bone marrow (n=8), and marrow alone (n=8) were evaluated. Twelve rats were euthanized at 11 days and 12 at 21 days. Explants were evaluated histologically for evidence of bone formation, and osteogenic activity was determined by an assay for alkaline phosphatase. Histological bone formation was absent in all groups at 11 days. At 21 days, in the Collagraft alone and bone marrow alone groups, no bone induction was noted. In contrast, 21-day specimens from the Collagraft plus bone marrow group showed newly formed bone. Alkaline phosphatase activity was negligible (<0.05 units/mg) in all 11-day specimens. Activity in Collagraft plus bone marrow specimens at 21 days (0.38 units/mg) was significantly higher than any other group (P<.001 for all comparisons). This study demonstrates that Collagraft plus bone marrow is an osteoinductive matrix. PMID- 12555840 TI - Medial clavicle physis fracture with posterior displacement and vascular compromise: the value of three-dimensional computed tomography and duplex ultrasound. AB - Fractures of the medial clavicle are rare, and most usually displace anterior or superior. Due to the structures in the mediastinum, accurate diagnosis of posterior fractures or dislocations is vital, but diagnosis can be difficult and delayed. Duplex ultrasound and three-dimensional computed tomography can be helpful in making the diagnosis. PMID- 12555841 TI - Intramuscular lipoma in an anomalous muscle belly of the middle finger lumbrical as a cause of carpal tunnel syndrome and trigger wrist. PMID- 12555842 TI - Bruns-Garland syndrome (diabetic amyotrophy). PMID- 12555843 TI - Traumatic lumbosacral nerve root avulsions in a pediatric patient. PMID- 12555844 TI - Early detection of Kienbock's disease with MRI treated by revascularization with a distal radius bone graft. PMID- 12555845 TI - Medial elbow pain in the throwing athlete. PMID- 12555846 TI - Introductory lecture: Time-resolved chemistry at atomic resolution. AB - Though time-resolved studies are still at an early stage, the field is rapidly being developed and applied to an increasingly broad spectrum of problems with timescales varying from seconds or more down to femtoseconds. In this overview a number of different techniques are discussed, with emphasis on chemical applications in which information is obtained at the atomic level. The need to correlate with theory, both for calibration of theoretical methods and to obtain related information not accessible experimentally, is stressed. PMID- 12555847 TI - The realization of sub-nanosecond pump and probe experiments at the ESRF. European Synchrotron Radiation Facility. AB - We present beamline ID09B that is designed for pump and probe experiments to 50 ps time-resolution. The beamline has been refurbished with a narrow-bandwidth undulator for Laue diffraction and diffraction from liquids. The new undulator has 235 poles, a 17 mm magnetic period and is operated at 6.5 mm gap. It produces a spectral flux of 2.0 x 10(8) photon/0.1% bw/pulse (10 mA) at the fundamental at 15.5 keV and an integral flux of 1.1 x 10(10) photon pulse(-1) in a 2.5% bandwidth. The optics has been renewed with a high-precision toroidal mirror and a cryogenic monochromator. The X-ray chopper used for single pulse selection is also described together with the femtosecond laser. Finally the diffraction from excited iodine molecules in CCl4 is investigated on the nanosecond time-scale. It turns out that the high-angle scattering is insensitive to the thermal chock from the laser: these oscillations are therefore readily used for structure determination. Conversely, the low-angle scattering probes the hydrodynamics of the liquid over longer length scales and the oscillations are believed to originate from thermal stress and expansion of the solvent. PMID- 12555848 TI - Femtosecond mid-infrared spectroscopy of condensed phase hydrogen-bonded systems as a probe of structural dynamics. AB - We report the first time-resolved site-specific mid-infrared study of the photo induced excited state hydrogen transfer reaction in 2-(2' hydroxyphenyl)benzothiazole (HBT) with 130 fs time resolution. The transient absorption of the C=O stretching band marking the keto*-S1-state appears delayed on a time scale of 30-50 fs after electronic excitation to the enol*-S1-state. Its line center subsequently shifts up by about 3-5 cm(-1) after excitation, depending on the excitation wavelength tuned between 315 and 349 nm. This effect is attributed to intramolecular vibrational energy redistribution (IVR) and vibrational energy relaxation (VER) processes. We observe for the first time the coherent effects of anharmonic coupling of low frequency modes (approximately 60 cm(-1), approximately 120 cm(-1)), on the C=O mode marking the product state. We ascribe the 120 cm(-1) mode to a Raman-active in-plane deformation mode that is coherently excited by the UV-pump pulse. We tentatively explain the coherent excitation of the infrared active 60 cm(-1) out-of-plane deformation mode by nonradiative processes within the excited enol state after electronic excitation. PMID- 12555849 TI - What we can learn about fast chemical processes from slow diffraction experiments. AB - The potential energy surface is an important determinant of a chemical reaction. Three ways of deducing non-trivial properties of such surfaces from the results of crystal structure analyses are discussed and illustrated with examples. (1) The mapping approach brings together structures of the same molecular fragment from different environments to outline reaction coordinates and vibrations. (2) Correlations between molecular structures and activation energies for a given reaction type reveal general and quantitative relations between seemingly independent entities such as ground state structure, force constants, reaction path length, activation energy and catalysis. (3) The evolution of atomic mean square amplitudes (displacement parameters) with temperature uncovers frequencies and atomic displacement patterns of large-amplitude vibrations in molecular crystals. Examples include the vibrations of molecular zeolite building blocks, the crankshaft motion of stilbenes, the dynamic coupling between pyramidal deformation of the amide NH2 group and hydrogen bonding, the bowl inversion of corannulenes and nucleophilic addition/elimination reactions. PMID- 12555850 TI - The frontiers of time-resolved macromolecular crystallography: movies and chirped X-ray pulses. AB - Three important frontiers of ultrafast time-resolved macromolecular crystallography are presented: extension of this technique to other biological systems; further developments in the elucidation of mechanism through the analysis of time-dependent movies to extract the underlying, time-independent, intermediate structures; and enhanced time resolution. The last is intimately linked with the nature of the pump-probe experiment itself, with the sources of random and, particularly, systematic experimental error, and with the factors that contribute to overall time resolution. All experiments to date have utilized the unchirped X-ray pulses that are emitted by synchrotron sources. Chirped pulses offer certain advantages for ultrafast X-ray experiments such as those based on Laue diffraction. An energy-chirped pulse maps photon energy into time; a Laue diffraction experiment maps photon energy into detector space. Hence, a Laue experiment with an energy-chirped pulse maps time into space. The proposed sub-picosecond photon source could provide an excellent source of intense, chirped hard X-rays for such experiments. PMID- 12555851 TI - Achieving photo-control of protein conformation and activity: producing a photo controlled leucine zipper. AB - We have recently developed a technique that has great potential in producing proteins with photo-control of conformation and consequently activity (J. R. Kumita, O. S. Smart and G. A. Woolley, Proc. Natl. Acad. Sci. U. S. A., 2000, 97, 3803-3808). The method is based on incorporating two cysteine residues into the sequence of a polypeptide. An azobenzene derivative is subsequently used to produce an intramolecular cross-link between the cysteine sulfhydryl groups. In previous work photo-isomerisation of the azobenzene moiety has been used to control the helicity of a monomeric peptide. In the experiments described here this method has been applied to the coiled coil leucine zipper peptide GCN4-p1. The aim was to produce a variant of GCN4-p1 whose helicity and consequently dimerisation is under direct photo-control. We have produced a modified GCN4-p1 incorporating two cysteine residues. The mutations introduced are shown to interfere with the ability of the uncross-linked peptide to form a coiled coil. After the peptide was cross-linked with the azobenzene derivative more normal coiled-coil behaviour was restored. Irradiation of the peptide producing a conformational change in the azobenzene cross-linker was accompanied by an increase in the helicity of the peptide. The work presented here highlights the potential of the use of photo-isomerisable cross-linkers to control protein activity through induced conformational change. In addition, the methodology has the potential to provide a fast trigger for the initiation of protein conformational changes. PMID- 12555852 TI - First investigations of the kinetics of the topochemical reaction of p-formyl trans-cinnamic acid by time-resolved X-ray diffraction. AB - Under UV irradiation p-formyl-trans-cinnamic acid (p-FCA) crystals in the beta phase dimerise irreversibly to solid 4,4'-diformyl-beta-truxinic acid. The experimental conditions were chosen in such a way (non-aqueous environment and room temperature) that the product formed is amorphous. The kinetics of this bimolecular reaction, which has not yet been characterised, was investigated by picosecond time-resolved X-ray diffraction. From the experimental results a mechanism for this topochemical reaction is proposed including two observed time constants, one less than 100 ps and another of several seconds. The feasibility of investigating this class of substances by time-resolved X-ray diffraction from third generation synchrotron sources and future free-electron lasers is discussed. PMID- 12555853 TI - Nanosecond time-resolved crystallography of photo-induced species: case study and instrument development for high-resolution excited-state single-crystal structure determination. AB - This work describes one of the first stages in the development of time-resolved photo-induced small-molecule single-crystal diffraction, whereby transient electron density perturbations, with lifetimes down to the nanosecond level, can be resolved at the atomic level. Knowledge of such ephemeral electronic effects is likely to yield key information regarding the origins of certain important physical properties, e.g. luminescent and non-linear optical effects, since it will allow the dynamics of electron density to be identified and quantified, and it is this that underpins such phenomena in a given molecule. The experimental methodology employs phase-locking pump-probe techniques such that the inherent time-structure of a synchrotron X-ray beam (nanoseconds) is harnessed and time gated in-phase with a femtosecond laser. The resultant beams, made coincident on the crystal in a periodic manner, and a diffraction pattern are recorded as a function of the Bragg angle, theta. Such technology is based upon the pioneering work carried out in sub-nanosecond time-resolved crystallography of macromolecular biological moieties (non-atomic resolution) at the ESRF, although one crucial difference here is the use of monochromatic irradiation and oscillatory motion rather than Laue 'snapshot' methodology, so that atomic resolution is possible. The experimental details of a case study conducted on ID9 at the ESRF, France, are described, whereby the feasibility of the excited-state structure determination of a luminescent rhenium carbene complex, [HNCH2CH2NHCRe(2,2'-bipyridine)(CO)3]Br, is realised. Key experimental parameters that are required for the success of such an experiment are discussed in the light of this study, together with other feasibility work conducted at the SRS, UK, and in the laboratory. Plans, designs and tests for the implementation of this technique in the UK, first at the SRS, and then at DIAMOND, the forthcoming UK synchrotron, are described, in particular with reference to the world-leading potential that DIAMOND could lend toward the development of this technique. PMID- 12555854 TI - Time-resolved and static-ensemble structural chemistry of hydroxymethylbilane synthase. AB - The enzyme hydroxymethylbilane synthase (HMBS, EC 4.3.1.8), 313 amino acid residues and MW 34 kDa, also known as porphobilinogen deaminase (PBGD), catalyses the stepwise polymerization of four molecules of porphobilinogen (PBG) to the linear tetrapyrrole 1-hydroxymethylbilane. Several crystallographic structures of HMBS have been previously determined, most recently including by time-resolved Laue protein crystallography of the Lys59Gln mutant form with reaction initiation undertaken by use of a flow cell carrying the substrate PBG. In this paper we review these structures and add new molecular graphics representations and analyses. Moreover we present a new structure refined at 1.66 A resolution using diffraction data recorded at cryo-temperature (100 K) in an attempt at trapping the polypeptide loop (residues 47 to 58) in the vicinity of the enzyme active site, missing in all previous structure determinations. This loop still has not appeared in the electron density maps, in spite of the advantage of cryo temperature, but nevertheless the 1.66 A cryo-structure extends the ensemble of known HMBS structures. The cryomodel of protein, cofactor and 320 bound water molecules has been refined to a final R-factor and R-free of 0.198 and 0.247 respectively; the PDB deposition codes, coordinates and structure factors are 1GTK and R1GTKSF respectively. Finally a protein comparison study is presented of the Mycobacterium tuberculosis (MTb) HMBS, with the E. coli HMBS. This has been done as preparation for future structural studies on the MTb HMBS from this important disease bearing organism. The overall amino acid sequence identity is 41%. Most interestingly there is a two-residue reduction in length of the loop referred to above (Asp 50 and Gly 58 being missing in the MTb form). This gives the hope that this loop will be less flexible and thus might become visible to crystallographic analysis. PMID- 12555855 TI - Structural dynamics of the receptor-binding domain of colicin E9. AB - Colicin E9 is a 61 kDa antibacterial protein secreted by E. coli. In order for it to enter the cytoplasm of susceptible bacteria and kill them by hydrolysing their DNA, the colicin must first interact with an outer membrane receptor on the target cell, BtuB, and a translocation pathway involving Tol proteins. The receptor binding, translocation and DNase functions of colicin E9 are housed in discrete structural domains, which have been independently expressed and characterized. The minimal receptor-binding domain is a 76 amino acid protein (min-R). X-ray structure determination of a related colicin shows its receptor binding-domain to have a helical hairpin structure (S. Soelaiman, K. Jakes, N. Wu, C. Li and M. Shoham, Molecular Cell. 2001, 8, 1053). Our solution NMR studies of min-R have confirmed it has a helical hairpin structure, and shown it has multiple slowly interchanging conformers and a flexible inter-helix loop. A plausible interpretation of these data is that in solution the helical hairpin can adopt a variety of structures differing in the spatial relationship of the two helices. A possible biological role for this involves the hairpin opening during translocation into bacteria. PMID- 12555856 TI - The dynamic transition in proteins may have a simple explanation. AB - The transition that has been observed in the dynamics of hydrated proteins at low temperatures (180-230 K) is normally interpreted as a change from vibrational, harmonic motion at low temperatures to anharmonic motions as the temperature is raised. It is taken to be an intrinsic property of proteins and has been associated with the onset of protein functions. Examination of the dynamic behaviour of proteins in solution within a defined timescale window suggests that certain observations can be explained without the need to invoke a discontinuity in the dynamics of proteins with temperature, i.e. the existence of a dynamical transition is not required. This is discussed in the context of recent evidence that enzyme activity is independent of the activation of anharmonic picosecond dynamics and declines steadily with temperature through the apparent dynamic transition, in accordance with the Arrhenius relationship. That similar timescale dependent dynamical behaviour has been observed experimentally in chain polymers, and seen also in computer simulations of silica glasses, suggests that the phenomenon may be of wide general relevance in both simple glassy and more complex polymeric systems. PMID- 12555857 TI - Mechanism of formation of DNA-cationic vesicle complexes. AB - Cationic vesicles and DNA form complexes that are promising gene delivery systems. Despite the increasing number of publications on their morphology and structure, the mechanism leading to their formation is not yet understood due to a lack of kinetic data. In the present study the kinetics of the interaction between DNA and cationic vesicles were followed using stopped-flow turbidity and small-angle neutron scattering techniques. The neutron real-time experiments were performed on a high-flux diffractometer, the D22 at the ILL, using a stopped-flow set-up. Extruded mixed vesicles of dimethyldioctadecylammonium bromide (DODAB) with various amounts of dioleoylphosphatidylethanolamine (DOPE) were investigated at 25 degrees C. The results show that the transition from unilamellar vesicles to a multilamellar structure upon DNA addition occurs in three steps. The first step, on the millisecond time scale, is currently not accessible to neutron scattering but was observed by stopped-flow turbidity and fluorescence experiments. The second step, on a time scale of seconds, corresponds to the formation of an intermediate with a locally cylindrical structure. As time progresses this unstable intermediate evolves to a multilamellar structure, on a time scale of minutes. An understanding of the mechanisms behind the DNA-cationic vesicle complex formation event will allow the production of more homogeneous, efficient delivery systems in pharmaceutically acceptable forms. PMID- 12555858 TI - Following the formation of nanometer-sized clusters by time-resolved SAXS and EXAFS techniques. AB - Time-resolved in situ SAXS and XAS measurements were carried out to monitor the formation of nanoparticles of the sulfides of cadmium and zinc, from solutions containing he corresponding acetate, and thioacetamide under solvothermal conditions. Analysis of the SAXS data shows that particles of ca 5 nm in radius form within the first few minutes of the reaction and then grow uniformly to ca 20 nm over a period of two hours resulting in a highly mono-dispersed particle distribution. EXAFS data of the CdS particles also prepared by solvothermal methods and recorded at 20 K, support the formation of nano-meter sized particles. PMID- 12555859 TI - Application of stopped flow techniques and energy dispersive EXAFS for investigation of the reactions of transition metal complexes in solution: activation of nickel beta-diketonates to form homogeneous catalysts, electron transfer reactions involving iron(III) and oxidative addition to iridium(I). AB - Stopped-flow techniques of rapid mixing have been combined with energy dispersive X-ray absorption spectroscopy to monitor the reaction of Ni(dpm)2 [dpm = Bu1C(O)CHC(O)Bu1] by aluminium alkyls (AlEt2X, X = OEt and Et) to form the active species for the catalytic di- and tri-merisation of hex-1-ene. Acquisition times down to ca. 30 ms were achieved on Station 9.3 of the SRS using a photodiode array detector. The EXAFS features of the resulting solution complexes are of the form [Ni(O-O)R)(alkene)]. In the presence of PPh3, [Ni(O-O)(R)(PPh3) appears to be the redominant type of species. The reduction of aqueous Fe(III) by hydroquinone was investigated on ID24 at the ESRF by Fe K-edge energy dispersive EXAFS with a CCD camera as detector, spectra were obtained in 1 ms or longer. No intermediate inner sphere complex was detected prior to the formation of aqueous Fe(II). Finally the oxidative addition of CH3SO3CF3 to [IrI2(CO)2]- was monitored on Station 9.3 with a silicon microstrip detector. A single acquisition of 400 micros was feasible, with spectra recorded in multiples of 1.2 ms. In that time, the first stage of the reaction had been completed, with a slower stage thereafter. The results are consistent with the two-stage ionic oxidative addition mechanism. PMID- 12555860 TI - Direct dynamics calculations of reaction rate and kinetic isotope effects in enzyme catalysed reactions. AB - Direct dynamics calculations employing hybrid quantum mechanical and molecular mehanical (QM/MM) potentials and molecular dynamics simulation methods have been used to explore the important dynamic role that enzyme structure has on proton transfer in the C-H bond breakage of a methylamine substrate by methylamine dehydrogenase (MADH). Canonical variational transition state theory with optimised multidimensional tunnelling corrections has been used to predict deuterium kinetic isotope effects corresponding to a range of enzyme conformations and to show the importance of donor acceptor separation, and transition state and product stabilisation within the active site. Large kinetic isotope effects can be predicted for proton transfer with both semi-empirical and ab initio electronic structure methods. PMID- 12555861 TI - Time-resolved computational protein biochemistry: solvent effects on interactions, conformational transitions and equilibrium fluctuations. AB - Solvent plays an important role in modulating internal motions of proteins. Here we present a computational method for including solvent effects on charge-charge interactions and on pathways between functional protein conformations, and examine solvent effects on equilibrium internal fluctuations in proteins. A computationally efficient charge reparametrisation method is presented that satisfactorily reproduces the electrostatic interactions present in a full continuum Poisson-Boltzmann representation. The application of charge reparametrisation in the calculation of a large-scale conformational transition pathway in a protein, annexin V, is illustrated. We also examine solvent effects on fast (picosecond timescale) internal protein dynamics. Nose-Hoover dual heatbath molecular dynamics simulations are performed. These simulations allow the solvent region to be fixed at one temperature and the protein at another. The results of the Nose-Hoover simulations on hydrated myoglobin confirm that the solvent temperature strongly influences the protein fluctuations. We consider to what extent the solvent can be considered to determine the high temperature protein dynamics. PMID- 12555862 TI - Theoretical studies of time-resolved spectroscopy of protein folding. AB - Recently, we have made significant improvements in the accuracy of calculations of the circular dichroism of proteins from first principles. The quality of these calculations (especially at 220 nm, a key wavelength, where the intensity of the band correlates well with the helical content of polypeptides) has given us confidence to use such calculations to analyse nanosecond molecular dynamics simulations of the folding of polypeptides. We use this combined approach to explore the influence of dynamics on the circular dichroism spectroscopy of polypeptides. We apply it to equilibrium molecular dynamics simulations of two beta-sheet proteins with similar structures, but differing circular dichroism spectra. We analyse a molecular dynamics simulation of the acid-unfolding of myoglobin. For both alpha-helical and beta-sheet conformations, we find that changes in dihedral angles of 30 degrees can change intensities of bands in circular dichroism spectra by up to 5000 degree cm2 dmol(-1). Thus, in isolation, moderate differences in circular dichroism spectra cannot be interpreted uniquely in terms of conformational changes. Examination of individual structures allows us to dissect the influence of conformation on the calculated circular dichroism spectra. Our results are aimed at providing a deeper understanding of the optical properties of proteins. An atomic level connection between molecular dynamics simulations and optical spectroscopy is increasingly desirable as theoretical and experimental studies begin to probe protein folding events reliably on the nanosecond timescale. PMID- 12555863 TI - Chemistry, physics and time: the computer modelling of glassmaking. AB - A decade or so ago the remains of an early flat glass furnace were discovered in St Helens. Continuous glass production only became feasible after the Siemens Brothers demonstrated their continuous tank furnace at Dresden in 1870. One manufacturer of flat glass enthusiastically adopted the new technology and secretly explored many variations on this theme during the next fifteen years. Study of the surviving furnace remains using today's computer simulation techniques showed how, in 1887, that technology was adapted to the special demands of window glass making. Heterogeneous chemical reactions at high temperatures are required to convert the mixture of granular raw materials into the homogeneous glass needed for windows. Kinetics (and therefore the economics) of glassmaking is dominated by heat transfer and chemical diffusion as refractory grains are converted to highly viscous molten glass. Removal of gas bubbles in a sufficiently short period of time is vital for profitability, but the glassmaker must achieve this in a reaction vessel which is itself being dissolved by the molten glass. Design and operational studies of today's continuous tank furnaces need to take account of these factors, and good use is made of computer simulation techniques to shed light on the way furnaces behave and how improvements may be made. This paper seeks to show how those same techniques can be used to understand how the early Siemens continuous tank furnaces were designed and operated, and how the Victorian entrepreneurs succeeded in managing the thorny problems of what was, in effect, a vulnerable high temperature continuous chemical reactor. PMID- 12555864 TI - A SAXS/WAXS XAFS study of crystallisation in cordierite glass. AB - New Cr X-ray absorption fine structure (XAFS) data have been combined with the results of small angle X-ray scattering (SAXS) and wide angle X-ray scattering (WAXS) experiments to probe in detail the crystallisation mechanism in cordierite (Mg2Al4Si5O18) glass doped with 0.34 mol% Cr2O3. By direct comparison with chromo aluminate spinels (MgCr2xAl2(1 - x)O4) Cr XAFS is used to determine the composition of the devitrified Cr species. This is identified as MgCr(0.18)Al(1.82)O4, which can be directly related to the Cr content in the starting glass and as a result the total crystalline volume in the fully developed ceramic is predicted to be 4%. In situ WAXS not only reveals the presence of the spinel phase but also a silica-rich stuffed quartz phase. This grows independently of the spinel and is probably nucleated from the glass surface. From our knowledge of the compositions of both crystalline phases we are able to deduce that the SAXS contrast between the surrounding glass and the spinel crystallites is 30 times greater than that between the quartz crystallites and the glass matrix, and therefore dominates the measured scattered intensity and the SAXS invariant that is derived from it. As a consequence we are able to show that the spinel crystalline volume fraction inherent in the SAXS is in close agreement with the 4% value obtained from the Cr XAFS. Furthermore in situ SAXS reveals the gradual development of the spinel particle size and shape during heat treatment. This is conducted in the super-cooled region just above the glass transition temperature, Tg. By employing a two-step annealing process nucleation can be separated from growth and from time-resolved SAXS measurements the alumino chromate nanocrystals are found to be closely monodispersed. Over a total time course of 600 min they grow from rough crystallites to smooth spherical particles of radius 21 +/- 2 nm, with a final density of (1.2 +/- 0.4) x 10(21) m(-3). As the process of ceramic formation takes place in the viscous melt, growth is indeed found to be limited by diffusion and is complete when all the Cr is exhausted. We use this comprehensive in situ study of crystallisation in cordierite glass to demonstrate the advantages of combining SAXS, WAXS and XAFS for probing the time-resolved chemistry, the microstructure and its development from nucleation sites, that underpins the processing of nanoparticle ceramics. PMID- 12555865 TI - Excited state molecular structure determination in disordered media using laser pump/X-ray probe time-domain X-ray absorption spectroscopy. AB - Advances in X-ray technologies provide opportunities for solving structures of photoexcited state molecules with short lifetimes. Using X-ray pulses from a modern synchrotron source, the structure of a metal-to-ligand-charge-transfer (MLCT) excited state of Cu1(dmp)2+ (dmp = 2,9-dimethyl-1,10-phenanthroline) was investigated by laser pump/X-ray probe X-ray absorption fine structure (LPXP XAFS) in fluid solution at room temperature on a nanosecond time scale. The experimental requirements for such pump probe XAFS are described in terms of technical challenges: (1) conversion of optimal excited state population, (2) synchronization of the pump laser pulse and probe X-ray pulse, and (3) timing of the detection. Using a laser pump pulse for the photoexcitation, a photoluminescent MLCT excited state of Cu1(dmp)2(BArF), (dmp = 2,9-dimethyl-1,10 phenanthroline, BArF = tetrakis(3,5-bis(trifluoromethylphenyl)borate) with a lifetime of 98 +/- 5 ns was created. Probing the structure of this state at its optimal concentration using an X-ray pulse cluster with a total duration of 14.2 ns revealed that (1) a CuII center was generated via a whole charge transfer; (2) the copper in the MLCT state bound an additional ligand to form a penta coordinate complex with a likely trigonal bipyramidal geometry; and (3) the average Cu-N bond length increases in the MLCT excited state by 0.07 A. In contrast to previously reported literature, the photoluminescence of this pentacoordinate MLCT state was not quenched upon ligation with the fifth ligand. On the basis of experimental results, we propose that the absorptive and emissive states have distinct geometries. The results represent X-ray characterization of a molecular excited state in fluid solution on a nanosecond time scale. PMID- 12555866 TI - Recent results from the in situ study of hydrothermal crystallisations using time resolved X-ray and neutron diffraction methods. AB - We present new time-resolved powder diffraction data measured in situ during the hydrothermal crystallisation of two families of crystalline inorganic materials. In the first study, we have used time-resolved energy-dispersive X-ray diffraction (EDXRD) to follow the formation of zeolitic zinc phosphates from amine phosphates and zinc oxide in acidic solutions at 60-150 degrees C. The advantage of this method is the ability to penetrate a laboratory-sized reaction vessel and to measure data in short (< 1 min) time intervals. Integration of the Bragg peak intensities during the crystallisation of the product allows accurate crystallisation curves to be produced. In addition, in a number of cases, we observe the formation of transient crystalline intermediate phases which can be identified by use of a new three-element detector that allows a large amount of diffraction data to be measured during the experiment. We are thus able to show that three-dimensional zinc phosphate architectures often form via low dimensional chain and layered phases, which is consistent with a recent aufbau model proposed for their formation. In the second study, we focus on the hydrothermal formation of ferroelectric barium titanate from TiO2 and barium salts in alkaline solution using time-resolved neutron diffraction. Although the time resolution of the neutron diffraction experiment is lower than the EDXRD experiment (data are measured in intervals of 5 min), we are able to penetrate reaction mixtures that are highly absorbing towards X-rays, and thus can measure data in a large volume reaction cell. Neutron diffraction data were collected on one of the highest-flux/highest detector-coverage diffractometers currently available; the GEM diffractometer at ISIS, UK. These experiments reveal that BaTiO3 crystallises after a large amount of TiO2 has been consumed; this implies that a dissolution crystallisation mechanism predominates. Additional mechanistic information is inferred by the observation of transient crystalline phases under certain reaction conditions. PMID- 12555867 TI - Are metastable, precrystallisation, density-fluctuations a universal phenomena? AB - In-situ observations of crystallisation in minerals and organic polymers have been made by simultaneous, time-resolved small angle X-ray scattering (SAXS) and wide angle X-ray scattering (WAXS) techniques. In isotactic polypropylene slow quiescent crystallisation shows the onset of large scale ordering prior to crystal growth. Rapid crystallisations studied by melt extrusion indicate the development of well resolved oriented SAXS patterns associated with long range order before the development of crystalline peaks in the WAXS region. Block copolymers self-assemble into mesophases in polymer melts above a critical chain length (or above a critical temperature) and this self-assembly process is shown to be susceptible to an incipient crystallisation. Mesophase formation is observed at anomalously high temperatures in ethylene-oxide containing block copolymers below the normal melting point of the polyoxy ethylene chains. Formation of calcium carbonate from aqueous solutions of sodium carbonate and calcium nitrate is observed to be a two-stage process and precipitation proceeds by the production of an amorphous metastable phase. This phase grows until it is volume filling and leads to the formation of the two polymorphs Calcite and Vaterite. These three sets of results suggest pre-nucleation density fluctuations, leading to a metastable phase, play an integral role in all three classes of crystallisation. In due course, this phase undergoes transformation to "normal" crystals. PMID- 12555868 TI - In situ neutron diffraction studies of single crystals and powders during microwave irradiation. AB - Microwave dielectric heating has become an important method in chemical synthesis and materials processing over the past 15 years, and in the case of the reactions in solutions, there is a well-developed understanding of heating mechanisms and their influence on reaction rate. In the solid-state however, there is much less clarity, despite the advantages to be gained from better insight into the way in which such electromagnetic radiation may couple directly to charge carriers, accelerating reactions in good conductors. The related issue of the influence of microwave irradiation on biological systems, in particular, proteins, and the way in which this may pose hazards to health is similarly poorly understood despite the obvious relevance this may have to the current debate on the influence of electromagnetic radiation, in particular, microwave transmission, on human health. One reason for the paucity of fundamental insight in both fields is because most work has been performed with microwave equipment whose design is derived from that of a domestic oven, and which is not ideal for in situ studies of microwave driven processes. We have been developing new methods of irradiating a variety of solid samples while measuring structural parameters through a range of diffraction techniques, and describe apparatus that will enable X-ray or neutron scattering measurements to be performed on powders or single crystals under microwave irradiation with controlled power level. We also describe preliminary studies of a single crystal of the molecular solid aspirin, and a powder of the microwave-susceptible ionic material BaTiO3, during microwave irradiation. PMID- 12555869 TI - Concluding remarks: Time-resolved chemistry: from structure to function. A summary. PMID- 12555870 TI - Recruitment of patients to clinical trials: an ambitious undertaking for the UK. PMID- 12555871 TI - Recruitment of patients to clinical trials: an ambitious undertaking for the UK. PMID- 12555872 TI - Steroids, radiotherapy, chemotherapy and stents for superior vena caval obstruction in carcinoma of the bronchus: a systematic review. AB - OBJECTIVE: To conduct a systematic review to determine the relative effectiveness of treatments currently employed in the management of superior vena caval obstruction (SVCO). SEARCH STRATEGY: Electronic searching of the Cochrane Clinical Trials Register, Medline and Embase with identification of further studies from references cited in trials identified by electronic searching. SELECTION CRITERIA: Both randomized and non-randomized controlled trials in which patients with carcinoma of the bronchus and SVCO had been treated with any combination of steroids, chemotherapy, radiotherapy or insertion of an expandable metal stent. DATA COLLECTION AND ANALYSIS: There were three randomized and 98 non randomized studies of which two and 44 respectively met the inclusion criteria. MAIN RESULTS: Superior vena caval obstruction was present at diagnosis in 10.0% of patients with small cell lung cancer (SCLC) and 1.7% of patients with non small cell lung cancer (NSCLC). In one small randomized trial in SCLC, the rate of SVCO relapse was not significantly reduced by giving radiotherapy on completion of chemotherapy. In another, in NSCLC, the addition of induction chemotherapy to a course of synchronous chemo-radiotherapy did not provide greater relief of SVCO. In SCLC chemotherapy and/or radiotherapy relieved SVCO in 77%; 17% of those treated had a recurrence of SVCO. In NSCLC, 60% had relief of SVCO following chemotherapy and/or radiotherapy; 19% of those treated had a recurrence of SVCO. Insertion of an SVC stent relieved SVCO in 95%; 11% of those treated had further SVCO but recanalization was possible in the majority resulting in a long-term patency rate of 92%. Morbidity following stent insertion was greater if thrombolytics were administered. REVIEWERS' CONCLUSIONS: Chemotherapy and radiotherapy are effective in relieving SVCO in a proportion of patients whilst stent insertion may provide relief in a higher proportion and more rapidly. The effectiveness of steroids and the optimal timing of stent insertion (whether at diagnosis or following failure of other modalities) remain uncertain. PMID- 12555873 TI - Dose-escalation with CHARTWEL (continuous hyperfractionated accelerated radiotherapy week-end less) combined with neo-adjuvant chemotherapy in the treatment of locally advanced non-small cell lung cancer. AB - PURPOSE: a radiation-dose-escalation study was undertaken to assess the therapeutic benefit of combining accelerated hyperfractionated radiotherapy (RT) with neo-adjuvant chemotherapy (CT) in non-small-cell lung cancer (NSCLC). MATERIALS AND METHODS: One hundred and thirteen patients with locally advanced NSCLC were entered into a phase II trial of CHARTWEL (CHART Week-End-Less) 54 Gy or 60 Gy with or without three cycles of CT. Acute oesophageal reactions and analgesia were scored for up to 8 weeks after the start of RT. Pneumonitis, lung fibrosis, spinal cord and oesophageal strictures, were assessed using clinical and radiological criteria from 3 months onwards and throughout the study. Haematological and gastrointestinal toxicity was monitored in those patients undergoing CT. Endpoints for treatment outcome were overall survival, disease free survival and loco-regional control. RESULTS: Chemotherapy enhanced the incidence and duration of acute dysphagia,but the increase was transient. Healing occurred in all cases and there has been no evidence of long-term oesophageal complications. Clinically, almost 25% of those receiving CT+RT had Grade 2 pneumonitis, higher than seen with RT alone. However, the 1 patient with severe Grade 3 pneumonitis was in the RT 60 Gy alone group. An incidence of 17% Grade 2 pulmonary fibrosis at 2 years was seen with CT, slightly lower than with RT alone. To date, there is no evidence of Grade 3 lung fibrosis. There was a higher scoring of lung damage with the radiological endpoint, which gave no indication that CT increased pulmonary toxicity over that of RT alone. Loco-regional control at 2 years was 37% and 55% for CHARTWEL 54 Gy and 60 Gy alone compared with 72% in those treated with 60 Gy and neo-adjuvant CT However, this did not translate into a survival advantage. CONCLUSIONS: This study of CHARTWEL combined with induction chemotherapy, has shown that the strategy is feasible and that a possible therapeutic benefit may be obtained by the addition of CT. Although neo adjuvant treatment increased acute mucosal reactions and slight-to-moderate pneumonitis seen with CHARTWEL 60 Gy, the clinical management and quality of life of these patients is similar to those treated with radiotherapy alone. PMID- 12555874 TI - Fatal pulmonary fibrosis associated with induction chemotherapy with carboplatin and vinorelbine followed by CHART radiotherapy for locally advanced non-small cell lung cancer. AB - In an attempt to evaluate the safety and efficacy of chemotherapy and continuous hyperfractionated radiotherapy (CHART) for non small cell lung cancer (NSCLC), a dose-escalation study was initiated, in which patients were treated with a combination of Vinorelbine and Carboplatin chemotherapy and CHART radiotherapy. The first cohort of 3 patients were treated with induction chemotherapy (Vinorelbine 30mg/m2 weeks 1,2,4 and 5, and Carboplatin, AUC = 5mg/ml/min weeks 1 and 4) followed by CHART radiotherapy (5400cGy in 36 fractions in 12 days on weeks 7 and 8). It was intended to then treat subsequent cohorts of patients with increasing doses of chemotherapy concomitantly with CHART, but unfortunately 2 of the first 3 patients both developed respiratory failure due to widespread pulmonary fibrosis, and died 7 and 9 weeks after completing treatment. At this point the study was closed. The combination of chemotherapy and CHART for NSCLC may have significant pulmonary toxicity and this potentially serious adverse effect needs to be carefully considered when planning future research studies is this area. PMID- 12555875 TI - Outpatient intensive chemotherapy for small cell lung cancer: five years experience of modified 'ICE' ifosfamide carboplatin and etoposide. AB - INTRODUCTION: Small Cell Lung Cancer (SCLC) is increasingly being treated in the district general hospital setting. The search for an active regimen which can be given with the least toxicity and best outcomes led us to use a modified ICE (Ifosfamide, Carboplatin and Etoposide) regimen and modify it further by using oral mesna instead of intravenous mesna. METHOD: All patients selected to receive the modified ICE regime over a 5-year period were included in our study. All patients were assessed for performance status and prognostic factors. Only those with WHO performance status 0-1 and Manchester prognostic score 0-3 were considered for ICE chemotherapy. All patients were followed up for 1 year after recruitment was completed. RESULTS: Median survival for all 32 patients was 18.4 months (CI 12.2-24.6) and for the 28 patients with limited disease the median survival was 19.9 months (CI 8.2-31.6). Toxicity levels were low with no neutropenic deaths. One patient died three days after treatment was started due to disease progression. A total of 6 patients remained alive one year after recruitment was completed. Five out of the 6 were followed up for at least 2 years. CONCLUSION: Using this out-patient modified ICE regime we have achieved a median survival comparable to other active chemotherapy regimes for SCLC with no significant increase in toxicity. PMID- 12555876 TI - From CHART to CHARTWEL in non-small cell lung cancer: clinical radiobiological modelling of the expected change in outcome. AB - CHART (Continuous Hyperfractionated Accelerated Radiotherapy) has been shown to improve the tumour control probability and survival relative to conventional radiotherapy in patients with inoperable non-small cell lung cancer (NSCLC). CHARTWEL (CHART Weekend-less) is a further development of this schedule escalating the physical dose to 60 Gy while maintaining the low dose per fraction of 1.5 Gy. In this schedule, three fractions, with a minimum interval of 6 h between fractions, are delivered 5 days per week. This extends overall treatment time from the 12 days of CHART to 18 days. Radiobiological modelling is used to estimate the expected tumour control and normal tissue morbidity after CHARTWEL relative to CHART. The estimations are based on the outcome of the CHART trial and published values for dose-fractionation and dose-response parameters for human tissues and tumours. Two new estimates of quantitative radiobiological parameters for early normal-tissue morbidity after chest irradiation are reported here. For radiation pneumonitis, the dose recovered per day is estimated at 0.44 Gy/day with 95% confidence limits 0.07 Gy/day and 0.80 Gy/day. For oesophagitis, the normalized dose-response gradient, gamma50, is estimated at 2.1 with 95% confidence limits 1.4 and 3.6. With regard to normal tissue effects, the increase in total dose when going from CHART to CHARTWEL is moderated by the slightly longer overall treatment time in case of early morbidity while the introduction of the weekend gaps may moderate the effect for late-responding normal tissues with a long repair halftime. Tumour control at 3 years is expected to increase by some 7-14 percentage points (from 19% to 26-33%) whereas the incidence of moderate and severe early oesophagitis and pneumonitis is expected to increase by about 2 percentage points. The incidence of late morbidity, lung fibrosis and oesophageal strictures, is expected to increase by 3-4 percentage points. The analyses conclude that CHARTWEL is likely to improve the therapeutic ratio relative to CHART. PMID- 12555877 TI - Practical methods for compensating for missed treatment days in radiotherapy, with particular reference to head and neck schedules. AB - Unscheduled interruption of a radiotherapy treatment can lead to significant loss in local tumour control, particularly in tumours that repopulate rapidly. General guidelines for dealing with such treatment gaps have been issued by the Royal College of Radiologists and more specific advice on the use of compensation methods has been published previously [Hendry et al., Clin Oncol 1996;8:297-307; Slevin et al., Radiother Oncol 1992;24:215-220]. This article further elaborates on the practical application of these methods. It sets out the main considerations arising in the especially critical case of head and neck treatments and simple calculations are used to illustrate the approaches which may be adapted for particular situations. Radiobiological parameter values are suggested for use in the calculations, but these may require modification in the light of further research in this important area. PMID- 12555878 TI - A basic treatment equivalent for high-dose-rate gynaecological brachytherapy--a pilot study. AB - BACKGROUND & PURPOSE: The aims of this pilot study were to assess the factors that were predictive of high-dose-rate (HDR) gynaecological brachytherapy duration and to model them using previously described Basic Treatment Equivalent (BTE) methodology. MATERIALS AND METHODS: This was a prospective single arm pilot study that aimed to enrol 20 patients from two centres. Patient, tumour and treatment factors were recorded. The duration of each component of brachytherapy was recorded. Univariate and bivariate analyses were conducted to identify factors that predicted overall brachytherapy duration. The generalized estimating equations method was used to derive an equation that predicted the duration of brachytherapy. RESULTS: Data were collected for 20 patients who underwent 53 episodes of brachytherapy, either as part of definitive radiotherapy (tandem and ovoids) or post-operatively (vaginal cylinder). Factors that were predictive of overall duration were technique (tandem and ovoids vs vaginal cylinder, P = 0.0007), treatment intention (definitive vs post-operative, P = 0.0001), type of plan (individual vs standard, P = 0.0001), hospital (1 vs 2, P = 0.0001) and body mass index (P = 0.0001). CONCLUSIONS: This study demonstrates the feasibility of examining factors that influence the duration of gynaecological brachytherapy using BTE methodology. To develop a reliable model, a larger multicentre study is needed. Such a model will allow comparisons of efficiency and more accurate assessment of treatment capacity between centres. PMID- 12555879 TI - An assessment of basic treatment equivalent at Cookridge Hospital. AB - The Australian model of Basic Treatment Equivalent (BTE) for measuring radiotherapy treatment productivity was developed for and tested within a busy UK radiotherapy centre and the results assessed against three other BTE models. The department's productivity was compared for three different models of treatment machine. Treatment fraction durations were timed for 362 consecutive treatment episodes on three treatment machines at Cookridge Hospital. Leeds. The treatment factors found to have a statistically significant impact on fraction duration were: initial fraction; patient mobility; number of fields; treatment intent and the type of treatment machine. In comparison with the Australasian study, shorter mean fraction times were shown for a number of types of treatment, carried out on one particular linac type, which had multileaf collimation (MLC). The average non operational time was 17% at Cookridge (31% in Australasia) with overall productivity gains. This efficiency may have been achieved partly by a higher number of staff per machine (five rather than four) and the presence of newer technology such as MLC. A BTE model was derived for Cookridge using the significant treatment factors and compared to other published models. The Cookridge model was most sensitive in predicting productivity. BTE models developed by other groups were also excellent in predicting treatment duration. The development of a BTE model is potentially applicable in the U.K. radiotherapy community at a time when optimizing activity per radiographer is key to minimizing waiting times. PMID- 12555880 TI - The development of a model of outpatient chemotherapy delivery--chemotherapy basic treatment equivalent (CBTE). AB - The aims of this study were to study the patient-, tumour- and treatment-related factors that significantly impact on treatment episode duration for outpatient chemotherapy treatment delivery, and to develop a new measure of outpatient chemotherapy throughput that considers variations in treatment duration compared with the older measures of patients treated per day. A pilot study in our institution randomly measured the duration of outpatient chemotherapy delivery for 266 occasions of service. Patient, tumour and treatment factors were collected and assessed for their impact on treatment duration using multivariate analysis. A new model of outpatient chemotherapy was developed using various modeling processes. Median treatment duration was 90 min. Significant factors that impacted on treatment duration were the chemotherapy regimen, type of infusion, patient age and whether the patients required a community nurse to be organized. A measure was developed (Chemotherapy Basic Treatment Equivalent or CBTE) that considers the variations in treatment duration and showed that although the daily number of patients treated in our department each day remained stable, there were wide fluctuations in workload when variations in treatment duration were considered. A new measure of chemotherapy workload has therefore been proposed although further testing across departments is required. If broadly implemented this could substantially improve resource planning, resource use and patient satisfaction as it considers variations in treatment duration, which is not previously considered in chemotherapy throughput statistics. PMID- 12555881 TI - Diagnostic radiology for radiotherapist: the case for structured training in cross-sectional imaging (CT and MRI). PMID- 12555882 TI - Positron emission tomography in the management of lymphomas. AB - Positron emission tomography is a functional imaging modality that capitalizes on biochemical changes within tumour cells to localize these changes within the body. As a functional imaging tool, unlike an anatomical imaging tool such as CT, it does not require enlargement of lymph nodes affected by disease but does require sufficient numbers of tumour cells to be present with altered biochemical function to visualize these disease sites. These changes are most commonly monitored using a glucose mimic fluorodeoxyglucose which is not only taken up into tumour cells but is trapped within these cells owing to alterations of the hexokinase and dephosphorylase enzymes. This review examines the current role of FDG PET imaging in patients with Hodgkins and Non-Hodgkins lymphoma and also speculates on future roles for this imaging modality. PMID- 12555883 TI - ICRU report 38: has the radiation oncology community accepted it? International Commission on Radiation Units & Measurements. PMID- 12555884 TI - Association of soft tissue sarcomas other than neurofibrosarcoma with neurofibromatosis. PMID- 12555885 TI - Chinese herbal remedies and radiotherapy. PMID- 12555886 TI - Primary ovarian carcinoid presenting as acute abdomen. PMID- 12555887 TI - Professor Jack Pearce, biographical note. PMID- 12555888 TI - Gordon memorial lecture. Poultry welfare: science or subjectivity? AB - 1. Most people in the developed world agree on what 'animal welfare' is, although it is impossible to give it a precise scientific definition. 2. The argument is made that animal welfare is all to do with the feelings of animals and not the primary needs that these feeling have evolved to protect. 3. Acceptance of subjective feelings as a legitimate subject for scientific investigation has a long and well-established history in science. This acceptance was interrupted by the rise of Behaviorism in the 20th century, but now seems to be re-established. 4. Subjective feelings cannot be studied directly. However, in the animal welfare debate, indirect evidence on feelings is extremely useful, and methods for obtaining this indirect evidence are described. 5. The poultry species are capable of feeling several states of suffering including fear, frustration and pain. A start has been made to elucidate these states and the conditions that cause them, but much remains to be done. Recent evidence suggests that the poultry species may also be capable of experiencing pleasure. 6. It is concluded that, although poultry welfare is all to do with the subjective feelings of the birds, it is possible to be objective and scientific about these feelings. Investigation into poultry welfare, therefore, really is science rather than subjectivity. PMID- 12555889 TI - Identification and quantification of granulocytes in caecal mucosa and submucosa of chickens experimentally infected with Eimeria tenella and Salmonella enteritidis. AB - 1. Poultry granulocytes are not clearly distinguished from each other with haematoxylin-eosin (HE) stain; thus, histochemical techniques must be used. Three experiments were carried out using 4-week-old Leghorn chickens. 2. Three, 80 chicken groups were orally infected with (1) 10(8) colony forming units (CFUs) Salmonella enteritidis, or (2) 10(4) Eimeria tenella oocysts, or (3) 10(8) CFUs S. enteritidis + 10(4) E. tenella oocysts. Ten chickens from each group were euthanased and caecum samples obtained. Caecum samples were fixed in 10% formalin (buffered, pH 7.4) at 4, 8, 12 h, 1, 3, 5, 7, and 14 d post-inoculation (PI). 3. Samples were stained using three different staining techniques: HE for the identification of heterophils and eosinophils, Ziehl-Neelsen for mast cells, and p-phenilenediamine dihydrochloride plus pyrocatechol (PPD + PC) for eosinophils. 4. Birds from Experiment 1 showed no changes in the numbers of granulocytes. Birds from Experiments 2 and 3 showed higher numbers of heterophils in caecal mucosa and submucosa separately, on d 5 and 7. In Experiment 3, a decrease was observed in submucosal mast cells on d 3. Chickens from Experiments 2 and 3 showed increased numbers of mucosal mast cells between d 7 and 14. 5. PPD + PC positively stained eosinophils, but not heterophils. 6. Numbers of heterophils and mast cells were increased during the acute inflammatory process caused by E. tenella. Therefore, mast cells could play a role as primary inflammatory cells. Eosinophils seem not to be part of the inflammatory process caused by E. tenella. PMID- 12555890 TI - Passage rate through the anterior digestive tract of broiler chickens fed on diets with ground and whole wheat. AB - 1. Birds were fed diets containing ground or whole wheat and titanium dioxide for 10 min, followed by dissection of 4 birds per treatment after 30, 60, 90, 120, 150 and 180 min. 2. The crop, proventriculus and gizzard, duodenum, jejunum, and ileum from each bird were dried and combusted, and titanium content in each section was determined. 3. Significant amounts of titanium were found in the small intestine of the birds as soon as 30 min after being given feed containing titanium, and significant amounts had passed the small intestine after 120 to 150 min. 4. Titanium contents equivalent to approximately 5 g feed were eliminated from the gizzard per hour independently of whether ground or whole wheat was used. Thus, a hypothesis that the use of whole grain would slow down passage rate through the upper digestive tract was not supported by the current results. PMID- 12555891 TI - The efficiency of nitrogen retention during compensatory growth of food restricted broilers. AB - 1. Two experiments were conducted to investigate the influence of compensatory growth, induced by early quantitative food restrictions, on the efficiency of nitrogen-retention in two lines of Ross broilers. 2. Birds were restricted to 80% of the ad libitum intake of the previous 24 h of the control group from d 4 to d 7. After the period of restrictions all birds were fed ad libitum. 3. In both trials, the final body weight of the restricted Ross 208(308) birds was similar or even higher than that of the control group. For the Ross 508 line, compensatory growth was substantial in the first trial, but non-existent in the second trial. In all cases, the mortality of restricted birds was lower than in control birds. 4. Not only environmental factors, but mainly chick quality, seemed to have a major influence on the capacity of the chickens to establish compensatory growth. 5. As compensatory growth was established, some improvement in N retention was induced. Although differences were not significant, they may be environmentally important. 6. It is concluded that a retardation of the early growth of fast growing broiler chickens can, in certain circumstances, reduce mortality and increase performance and N retention. PMID- 12555892 TI - Lysine requirement of growing male Pekin ducks. AB - 1. One growth experiment and one balance test were conducted to study the response to increasing levels of dietary lysine supplementation in male Pekin ducks with special reference to the growth periods from 1 to 3 weeks and 4 to 7 weeks of age. 2. Two different low-lysine diets were used as basal diets in both periods. The basal lysine levels were 7.6 g/kg (d 1 to 21) and 6.2 g/kg (d 22 to 49) and the ranges in lysine concentration were 7.6 to 12.6 g/kg (d 1 to 21) and 6.2 to 11.2 g/kg (d 22 to 49). 3. Growth performance, feed conversion efficiency and meat yield increased (P < 0.05) with increasing lysine concentration (requirement defined as 95% of the asymptote). 4. It is concluded that the dietary lysine concentration should be 0.93 g/MJ nitrogen corrected apparent metabolisable energy (AMEN) (11.7 g/kg) for the starter period (until d 21) and 0.75 g/MJ AMEN (10.0 g/kg) for the grower period (from d 22 onwards). PMID- 12555893 TI - Improvement of growth and nutritive value in chicks with non-genetically modified phytase product from Aspergillus niger. AB - 1. Non-genetically modified (non-GM) phytase product derived from Aspergillus niger possesses various side active enzymes including alpha-amylase, protease, cellulase and hemicellulase. In contrast, the product of genetically modified (GM) phytase product has much less side active enzyme since the capacity of phytase production is reinforced by gene modification. In the present study we have tried to determine whether the difference of side enzyme activity of phytase product affects growth performances and nutritive value in chicks; in addition we tried to characterise the physiological change induced by the difference of side active enzymes. 2. Single Comb White Leghorn male chicks at 7 d of age were fed on experimental barley-based diets for 10 d. The feeding trial was of a factorial design (3 x 2 x 2), having three types of dietary phytase products (control, non GM or GM phytase products derived from A. niger at 1000 U/kg diet), two levels of dietary available P supplement (0 or 6 g/kg diet) and two levels of dietary protein (CP 180 or 120 g/kg). 3. The non-GM phytase product caused a 6% increase in final body weight and feed efficiency compared with the control and the GM phytase product without interacting with dietary protein and available P level. However, in birds given available P-free diet, both non-GM and GM phytase products induced a 20% increase in plasma P concentration, suggesting no difference in phytase activity between the non-GM and GM phytase products. 4. The balance study showed that the metabolisable energy of the non-GM phytase product (15.6 +/- 0.05 kJ/g diet) was significantly higher among the treatments (control, 15.1 +/- 0.05; GM phytase product 15.3 +/- 0.07). The non-GM phytase product also increased the rate of food passage through the crop, and caused a drastic reduction in intestinal weight, perhaps as a consequence of digestion of non starch polysaccharides. 5. We conclude that the side active enzymes in non-GM phytase product improve growth performance and nutritive value of the diet in chicks. However, the efficacy of phytase activity should not be different between non-GM and GM phytase products. PMID- 12555894 TI - Influence of level of feed input and procedure on metabolisable energy and endogenous energy loss (EEL) with adult cockerels. AB - 1. Two experiments were performed to study the relationship between apparent metabolisable energy (AME) and true metabolisable energy (TME) values at different feed intakes. Rhode Island Red (RIR) adult cockerels were used in two bioassays, under standard conditions. 2. In the first experiment (precision feeding) the birds were not fed for 24 h and then various quantities of maize were fed. There were 6 replicates per treatment. Droppings voided during the 48 h of the experimental period were collected and assayed for gross energy and nitrogen. 3. In the second experiment, each bird was given a 3-d adaptation period, fasted for 24 h and then given maize as an admixture (conventional addition method: CAM) to the same rations as those used in the first experiment. During the 3 d of the experimental period, droppings were collected and frozen for chemical analysis. 4. The results showed that mean metabolisable energy (ME) values obtained by the precision-feeding procedure were higher than those obtained by CAM. The endogenous energy loss (EEL) (FEm + UEe) determined by precision feeding was lower than that given by CAM. 5. It was concluded that the method of feeding does not influence ME of maize at different feed inputs. However, differences among intercepts of the regression between ME and feed input suggest that EEL varies at different maize inputs. 6. The optimum maize quantity was approximately 35 to 40 g for adult RIR cockerels by both procedures. PMID- 12555895 TI - Effect of vitamin C on pulmonary hypertension and muscularisation of pulmonary arterioles in broilers. AB - 1. Three hundred and eighty 1-day-old Arbor Acres broilers were divided into control (A) and experimental (B, C, D, and E) groups. 2. After 14 d of age the experimental groups were subjected to a cool temperature challenge by lowering the temperature 1 to 2 degrees C per day down to 12 degrees C, and maintaining this temperature until 7 weeks of age. 3. At the same time, 1.5 mg/kg 3,3,5 triiodothyronine (T3) was added to the diet of groups D and E, and 500 mg/kg ascorbic acid (vitamin C) to the diet of groups C and E. 4. The incidence of pulmonary hypertension syndrome (PHS), body weight gain and feed intake were measured weekly. Lung and blood samples were collected weekly from 10 birds per group beginning on d 14, and the percentage of thick-walled peripheral lung vessels (% TWPV) and packed cell volume (PCV) were determined. 5. The lower ambient temperature and diets supplemented with T3 increased PHS incidence and % TWPV and decreased body weight gain. 6. There was an increase in PCV after 5 weeks of age under lower ambient temperature 3 and the PCV values 14 were also significantly increased by T3. 7. Vitamin C supplementation reduced PHS incidence and % TWPV but did not change packed cell volume, body, weight gain, feed intake, or feed conversion. 8. It is concluded that vitamin C reduced PHS and the associated muscularisation of pulmonary arterioles induced by exposing broilers to cool environmental temperatures and feeding them with T3. PMID- 12555896 TI - Gene expression of bone-associated cytokines in MG63 osteoblast-like cells incubated with acrylic bone cement extracts in minimum essential medium. AB - This study examined the effects of in vitro challenge with four polymerized acrylic bone cements (Sulfix-60, CMW 1, CMW 2, and CMW 3) on the expression of interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and transforming growth factor-beta1 (TGF-beta1) mRNAs in the osteoblast-like cell line MG63. The extracts of the cements in minimal essential medium (MEM) were tested following 1 h and 7-day curing. A semi-quantitative analysis of the cytokine-specific mRNAs was carried out by agarose gel densitometry and expression was compared with the GAPDH housekeeping gene. The ratio between cytokine gene expression and GAPDH expression was calculated. The mRNA specific for the bone-resorbing cytokines IL 1beta and IL-6 was low in basal conditions. IL-1beta mRNA increased only after incubation with the extract of CMW 1 following 1-h curing. The mRNA specific for the bone-resorbing cytokine IL-6 also increased after contact with CMW 1 at both curing times. Sulfix-60 and CMW 3 following 7-day curing, but not after 1 h, induced higher levels of IL-6 mRNA than the control. CMW 2 after 1-h curing constantly determined the expression of IL-6 mRNA, but at low levels. The mRNA specific for TGF-beta1 was also expressed by the MG63 osteoblast-like cells in basal conditions. The levels increased after contact with Sulfix-60 after 7-day curing and with CMW 1 after 1-h curing. CMW 2 after 7-day curing decreased TGF beta1 mRNA. In conclusion, the highest expression of the cytokines IL-1beta, IL 6, and TGF-beta1 mRNA was determined by CMW 1. If the results are confirmed in vivo, the increased expression of the osteolytic cytokines induced by the bone cement might result in loosening of the prosthesis, even with all the restrictions of an in vitro study on continuous cell lines. PMID- 12555897 TI - A free volume microprobe study of water sorption in a contact lens polymer. AB - The kinetics of moisture uptake in a rigid gas-permeable contact lens material, Fluoroperm60, has been investigated. The free volume changes accompanied by the sorption of water were monitored using the positron lifetime spectroscopy (PLS) technique. The positron results indicated swelling in the initial stages of sorption and the filling of vacant free volume sites with water in the later stages of sorption. The sorption curve suggests that the diffusion process in this polymer follows the dual-mode sorption model. Using this model, it was possible to separate Fickian-controlled diffusion and relaxation-controlled diffusion. Furthermore, the positron results combined with the sorption results indicated that Fujita's free volume theory does not seem to be valid for the diffusion process in the present sample under study. The variation of the permeability as a function of the free volume suggests that the amount of water in a contact lens is a measure of its permeability. PMID- 12555898 TI - Mechanical properties and in vitro degradation of bioresorbable knitted stents. AB - The aim of this study was to characterize the mechanical properties and in vitro degradation of bioresorbable knitted stents. Each stent was knitted using a single self-reinforced fibre made out of either PLLA or 96L/4D PLA or 80L/20G PLGA. The mechanical and physical properties of the fibres and stents were measured before and after gamma sterilization, as well as during in vitro degradation. The mechanical properties of the knitted stents made out of bioresorbable fibres were similar to those of commercially available metallic stents. The knitting geometry (loop height) had a marked effect on the mechanical properties of the stents. The rate of in vitro degradation in mechanical and physical properties for the PLLA and 96L/4D PLA stents was similar and significantly lower than that of the 80L/20G PLGA stents. The 80L/20G PLGA stents lost about 35% of their initial weight at 11 weeks. At this time, they had lost all their compression resistance strength. These data can be used as a guideline in planning further studies in vivo. PMID- 12555899 TI - Effect of gamma, ethylene oxide, electron beam, and plasma sterilization on the behaviour of SR-PLLA fibres in vitro. AB - The aim of this study was to evaluate the effect of various sterilization processes on the physical and mechanical properties of self-reinforced bioabsorbable fibres made out of polylactide (PLLA). The samples were sterilized using plasma, ethylene oxide (one and two cycles), gamma (25 kGy at room temperature, 25 kGy in dry ice, and 2 x 25 kGy at room temperature), and electron beam (15, 25, and 55 kGy) sterilization. The intrinsic viscosity, crystallinity, and mechanical properties (modulus of elasticity, yield strength, and ultimate tensile strength) were tested before and immediately after each sterilization treatment, as well as up to 30 weeks in vitro. Compared with unsterilized fibres, the intrinsic viscosity was markedly decreased after radiation sterilization (gamma and electron beam) and the loss in mechanical properties was accelerated during in vitro degradation. Plasma and ethylene oxide (one and two cycles) did not markedly alter the properties of the samples after sterilization or during in vitro degradation. These data are important for determining the effect of various sterilization processes on the physical and mechanical properties of polylactide based materials and can be used to predict how fast degradation of the mechanical properties of the self-reinforced PLLA will occur. They can also be used to tailor the degradation kinetics to optimize implant design. PMID- 12555900 TI - Phase separation and physical properties of PEO-containing poly(ether ester amide)s. AB - Poly(ether ester amide) (PEEA) copolymers based on poly(ethylene glycol) (PEG), 1,4-butanediol, and dimethyl-7,12-diaza-6,13-dione-1,18-octadecanedioate (a diester-diamide monomer) were synthesized by a two-step polycondensation reaction. The obtained segmented copolymers are hydrophilic, with a water uptake of 24-340%. PEEA copolymers showed microphase separation, as observed by differential scanning calorimetry (DSC). The long spacing determined by small angle X-ray scattering shows an increase in the hydrophilic domain size with increasing PEO content. By varying the copolymer composition, the E-modulus of PEEA could be varied between 61 and 427 MPa, with tensile strengths ranging from 12 to 39 MPa. The elongation at break can reach values up to 850%. The mechanical properties decrease with the uptake of water. However, PEEAs with a relatively low content of PEO still retain good tensile properties and are, in principle, suitable for biomedical applications. PMID- 12555901 TI - In vivo biocompatibility of gelatin-based hydrogels and interpenetrating networks. AB - The in vivo host response to two gelatin-based hydrogel systems of varying crosslinking modalities and loaded with the anti-inflammatory agent dexamethasone sodium phosphate was investigated. Either gelatin was chemically crosslinked with glutaraldehyde, or polyethyleneglycol diacrylate was photopolymerized around gelatin to form interpenetrating networks. The subcutaneous cage implant system was utilized to determine differential leukocyte concentrations in the inflammatory exudate surrounding the materials as indices for biocompatibility and drug efficacy in vivo. Most of the crosslinked gelatin-based materials, either via glutaraldehyde fixation or interpenetrating network formation, elicited stronger inflammatory responses than either of the starting materials, gelatin and polyethyleneglycol diacrylate. In general, dexamethasone delayed and intensified the inflammatory response. The loss of material mass did not correlate directly with the degree of cellular inflammatory response, but increased with longer implantation time and decreased with more extensive fixation. PMID- 12555902 TI - Odds and ends: risk, mortality, and the politics of contingency. AB - Medical anthropology's cogent rethinking of conventional biomedical categories has largely overlooked the core problems of one key concept of both biomedical and social scientific analysis: risk. In particular, the use of the term in medical anthropology (and the social sciences more generally) frequently rests on two assumptions: (1) that contingency necessarily constitutes a threat to individual experience or social order; and (2) that a risk management paradigm that relies on a model of statistical probability is the ontologically preeminent way of engaging chance. Other approaches which do not take risk as the starting point for understanding contingency also have problems; they too assume that contingency is necessarily cause for crisis. These problematic root assumptions lead social analysts to miss how individual actors and local communities variously engage, rather than minimize, contingency. I suggest a new approach that instead aims to treat contingency as normatively neutral and as arising in four domains of experience. Conventional approaches also miss how attempts to account for unexpected events themselves involve struggles between competing paradigms (or tropes) of chance. This contest over accountability I call here the politics of contingency, and I seek thereby to signal the need to renovate our language of uncertainty in order to address its political dimensions. I trace the literature to identify some sources of these terminological problems, and through an examination of the life and death of a close contact in Chania, Crete, I explore his own approach to chance and the different, competing interpretations of his death. I thereby demonstrate the importance of revamping the conventional approach to understanding the contingent nature of human life. PMID- 12555903 TI - Between "science" and "superstition": moral perceptions of induced abortion among young adults in Vietnam. AB - Despite the increasing use of reproductive technologies the world over, anthropological studies have paid remarkably limited attention to the ethical dilemmas involved in people's choices of such technologies. Against this background, the author analyzes moral perceptions of induced abortion among unmarried young adults in urban North Vietnam. While the ethical aspects of abortion are shrouded in silence in public life in Vietnam, the young people participating in the study expressed strong moral scepticism towards the practice of abortion, even while undergoing it themselves. Through the analysis of young people's experiences and perceptions, the paper demonstrates how moral ideas are tied to particular social situations and structured by larger socio-political circumstances. It is argued that moral notions which are dominant in a society's public sphere may not be representative of the moral sentiments that are lived in practice and felt in private. PMID- 12555904 TI - Commercial sex work or ukuphanda? Sex-for-money exchange in Soweto and Hammanskraal area, South Africa. AB - This article introduces the concept of ukuphanda, a Zulu verb that is used to describe the sex-for-money exchanges that take place outside of commercial sex work in Soweto and Hammanskraal area, South Africa. In line with the ethnographic literature from others areas of sub-Saharan Africa, it is argued that women who exchange sex for money in taverns do not self-identify as commercial sex workers and experience less stigma from the community. Unlike commercial sex work (as characterized by the commercial sex work in Hillbrow, Johannesburg), which is understood to be associated with short skirts and other revealing attire, sex-for money exchange in the taverns is viewed as more private, ambiguous and informal. Women who work as informal sex workers, or "-phandela imali" ('try to get money'), are understood to be using sex-for-money exchange to survive financially. PMID- 12555905 TI - A tale of two policies: the French connection, methadone, and heroin epidemics. AB - The law enforcement and treatment policies of the Nixon administration are often credited with ending the epidemic of heroin addiction that rose in America's cities in the 1960s. In this article it is argued that although the interventions did in fact cause a major change in heroin distribution and use, the epidemic did not end in any simple way. The decline in heroin and increase in methadone that resulted from the Nixon policies lead to a shift for many addicts in both clinical and street settings from one narcotic to another. The temporary shortage of heroin that resulted from law enforcement was quickly compensated for with methadone, as well as with new distribution systems from Southeast Asia and Mexico. In the end, the interventions caused a change in an enduring "heroin system," a change that left that system in a stronger form in terms of supply and in a situation of continuing growth in terms of the number of addicts. PMID- 12555906 TI - An evaluation of contingency strength and response suppression. AB - Typically, functional analyses of severe problem behavior have been conducted in two ways: (a) The target response is reinforced immediately after it occurs, or (b) the target response is reinforced on some schedule thought to mimic a naturally occurring schedule. We evaluated the effects of contingency strength in reducing levels of problem behavior with 2 participants who had been diagnosed with developmental disabilities. Results showed that under a neutral contingency, one in which the probability of reinforcement for aggression was equal to the probability of reinforcement for the nonoccurrence of aggression, rates of aggression were suppressed to low levels for both participants. PMID- 12555907 TI - The influence of activity choice on problem behaviors maintained by escape versus attention. AB - This study assessed whether the function of an individual's problem behavior was related to the effectiveness of an intervention involving choice among tasks. Analogue functional analyses were conducted with 7 students with various diagnoses to determine whether problem behaviors were maintained by escape or attention. Following identification of the function of each student's problem behavior, reversal designs were used to assess the effectiveness of an intervention that allowed the students to choose their own instructional tasks. Results showed that students who displayed escape-maintained problem behavior showed substantial reductions in such behavior when they were provided with opportunities to choose among tasks. On the other hand, students who displayed attention-maintained problem behavior did not show any effects as a result of the choice intervention. These findings are discussed in terms of the effective use of behavior management programs involving choice and the reduction of problem behavior. PMID- 12555908 TI - An evaluation of two differential reinforcement procedures with escape extinction to treat food refusal. AB - Consumption of solids and liquids occurs as a chain of behaviors that may include accepting, swallowing, and retaining the food or drink. In the current investigation, we evaluated the relative effectiveness of differential reinforcement of the first behavior in the chain (acceptance) versus differential reinforcement for the terminal behavior in the chain (mouth clean). Three children who had been diagnosed with a feeding disorder participated. Acceptance remained at zero when differential reinforcement contingencies were implemented for acceptance or mouth clean. Acceptance and mouth clean increased for all 3 participants once escape extinction was added to the differential reinforcement procedures, independent of whether reinforcement was provided for acceptance or for mouth clean. Maintenance was observed in 2 children when escape extinction was removed from the treatment package. The mechanism by which consumption increased is discussed in relation to positive and negative reinforcement contingencies. PMID- 12555909 TI - Effects of strategic versus tactical instructions on adaptation to changing contingencies in children with adhd. AB - This study examined the effects of two types of instructions on the academic responding of 4 children with attention deficit hyperactivity disorder. Tactical instructions specified how to distribute responding between two concurrently available sets of math problems associated with different variable-interval schedules of reinforcement. Strategic instructions provided a strategy to determine the best way to distribute responding. Instruction conditions were counterbalanced in an ABAB/BABA reversal design nested within a multiple baseline across participants design. Experimental sessions consisted of a learning session in which participants were provided with one type of instruction, followed by a test session in which no instruction was provided. The schedules of reinforcement were subsequently reversed during test sessions. When learning and test schedules were identical, the responding of all 4 participants closely matched the reinforcement schedules. When tactical instructions were provided and schedules were subsequently changed, responding often remained under the control of the instructions. When strategic instructions were provided, responding more quickly adapted to the changed contingencies. Analysis of postsession verbal reports indicated correspondence between the participants' verbal descriptions (whether accurate or inaccurate) and their nonverbal patterns of responding. PMID- 12555910 TI - The effects of differential and lag reinforcement schedules on varied verbal responding by individuals with autism. AB - Variability has been shown to be a reinforceable dimension of behavior. One procedure that has been demonstrated to increase variability in basic research is the lag reinforcement schedule. On this type of schedule, a response is reinforced if it differs from a specified number of previous responses. Lag schedules are rarely used, however, for increasing response variability in applied settings. The purpose of the present study was to investigate the effects of a lag schedule of differential reinforcement on varied and appropriate verbal responding to social questions by 3 males with autism. A reversal design with a multiple baseline across subjects was used to evaluate the effects of the lag schedule. During baseline, differential reinforcement of appropriate responding (DRA) resulted in little or no varied responding. During the intervention, a Lag 1 requirement was added to the DRA (Lag 1/DRA) resulting in an increase in the percentage of trials with varied and appropriate verbal responding for 2 of the 3 participants. In addition, an increase in the cumulative number of novel verbal responses was also observed for the same 2 participants. These results are discussed in terms of reinforcement schedules that support variability, generalization, and potential stimulus control over varied responding. PMID- 12555911 TI - Sensitivity of children's behavior to probabilistic reward: effects of a decreasing-ratio lottery system on math performance. AB - Probabilistic reward has been shown to affect children's choice behavior in game like activities. We examined the effects of a lottery system containing progressively lower exchange ratios on children's completion of math problems. Two of the 3 children completed problems above baseline levels until the chance of exchange dropped to 25%. This study describes a potentially useful method for examining reinforcement schedules in applied settings and extends previous research on probabilities of reinforcement. PMID- 12555912 TI - The effects of noncontingently available alternative stimuli on functional analysis outcomes. AB - The effects of noncontingently available alternative stimuli on functional analysis outcomes were evaluated for 2 individuals. Results suggested that noncontingent access to preferred items resulted in reduced levels of attention maintained behavior. The results are interpreted in terms of establishing operations. PMID- 12555913 TI - Reinforcing efficacy of food on performance during pre- and postmeal sessions. AB - Assessments were conducted to identify preferred foods for 9 adults with severe to profound mental retardation. Subsequently, the reinforcing effects of each participant's most highly preferred food were evaluated during sessions conducted before and after lunch. Results showed that 4 participants' response rates were higher during premeal sessions than during postmeal sessions. By contrast, pre- and postmeal response rates were indistinguishable for the other 5 participants. These results indicate that the reinforcing efficacy of food may (but does not necessarily) diminish following meals and suggest that the influence of meal schedules should be examined on an individual basis when food is used as reinforcement during training sessions. PMID- 12555914 TI - Shaping exhale durations for breath CO detection for men with mild mental retardation. AB - Roll, Higgins, and Badger (1996) used a carbon monoxide (CO) detector to determine whether participants smoked in a smoking-cessation study. We sought to replicate their work with adults with mild mental retardation. However, verbal instructions were inadequate to establish stable exhalations of sufficient durations for reliable and accurate CO evaluation. This report describes a shaping procedure that enabled 3 of 4 participants to achieve 20-s exhalation durations. PMID- 12555915 TI - Effects of mand-tact versus tact-only training on the acquisition of tacts. AB - We sought to replicate and extend Carroll and Hesse's (1987) study of the acquisition of tacts by including participants with and without developmental disabilities. As in Carroll and Hesse, the present results showed that mand-tact training, rather than tact-only training, led to more rapid acquisition of tacts. Tacting on follow-up tests did not differ. In addition, our results show that mand-tact training established both verbal operants involved about as rapidly as tact-only training established only one verbal operant. DESCRIPTORS: verbal behavior, tact acquisition, mand-tact training, tact-only training, follow-up PMID- 12555916 TI - Evaluating the function of applied behavior analysis a bibliometric analysis. AB - Analysis of scholarly citations involving behavioral journals reveals that, consistent with its mission, applied behavior analysis research frequently references the basic behavioral literature but, as some have suspected, exerts narrow scholarly influence. PMID- 12555917 TI - Feedback and its effectiveness in a computer-aided personalized system of instruction course. AB - In a computer-managed version of Keller's personalized system of instruction, students received frequent feedback from more advanced students within the course. Overall accuracy of student-provided feedback was 87%, and students complied with 61% of the feedback. PMID- 12555919 TI - On the status of knowledge for using punishment a commentary. PMID- 12555920 TI - Punishment happens: some comments on Lerman and Vorndran's review. AB - Some readers will view the article by Lerman and Vorndran as controversial. It is a review of existing basic and applied research on punishment and a call for additional research on punishment. The thesis of my commentary is that the paper should not be viewed as controversial. Punishment happens. To ignore a natural phenomenon and its implications for a technology of behavior is akin to ignoring the physical nature of the universe. A science and a technology of behavior are incomplete without research on punishment. Five reasons to pursue punishment research are discussed, along with some caveats. PMID- 12555921 TI - Punishment a primary process? PMID- 12555918 TI - On the status of knowledge for using punishment implications for treating behavior disorders. AB - In this paper, we review basic and applied findings on punishment and discuss the importance of conducting further research in this area. The characteristics of responding during punishment and numerous factors that interact with basic processes are delineated in conjunction with implications for the treatment of behavior disorders in clinical populations. We conclude that further understanding of punishment processes is needed to develop a highly systematic, effective technology of behavior change, including strategies for improving the efficacy of less intrusive procedures and for successfully fading treatment. PMID- 12555922 TI - Occlusal adjustment: quo vadis? PMID- 12555923 TI - Just what we need: another academy. PMID- 12555924 TI - AMA responds to Dr. Riley Lunn's editorial in the October 2002 issue of CRANIO. PMID- 12555925 TI - Patients as partners: comments of Dr. Uppgaard on Dr. Riley Lunn's Concepts editorial in October CRANIO. PMID- 12555926 TI - A randomized controlled trial of therapeutic exercise for clicking due to disk anterior displacement with reduction in the temporomandibular joint. AB - Therapeutic exercise is a new concept of treatment for patients with clicking due to anterior disk displacement with reduction (ADDWR). In order to investigate the efficacy of the exercise, we designed a randomized controlled clinical trial to compare with no-treatment controls. The subjects were patients who complained of painless unilateral reciprocal clicking and were diagnosed as having ADDWR by Magnetic Resonance Imaging (MRI). Forty-two patients were randomly assigned. Three months after the randomization, the success rate was 61.9% (13/21 joints) in the exercise group and 0% (0/21 joints) in the control group, with a significant difference between the two groups (p=0.0001). However, captured disks in the successful cases were seen in only 23.1% on MRI examination. In conclusion, the therapeutic exercise for clicking due to ADDWR is significantly effective in reducing the clicking, and thought to be much more conservative and cost-effective than splint therapy or surgery. PMID- 12555927 TI - A clinical comparison of internal joint disorders in patients presenting disk attachment pain: prevalence, characterization, and severity of bruxing behavior. AB - This study clinically documents the presence of disk-attachment pain (DAP) related to the severity of bruxism and to the prevalence of internal joint disorders in patients and controls. The sample consisted of 394 temporomandibular disorder (TMD) and bruxing behavior patients and 104 nonbruxer control subjects. The TMD group was subdivided in a group of 109 disk-attachment pain patients and 285 bruxing behavior-nonDAP groups. The second reference group was a sample of 104 nonbruxing behavior subjects. Comprehensive evaluations (including history of signs and symptoms), use of questionnaires, clinical examination, palpation of muscles and joints, classification of bruxing behavior by the degree of severity, and diagnostic tests were performed in patients and controls. There was a prevalence of disk-attachment pain in the group of TMD and bruxing behavior patients. The degree of jaw opening was lower in the DAP group as compared to the TMD/bruxism-nonDAP group and controls. The mean Visual Analog Scale (VAS) value for the joint pain complaints was slightly elevated in the DAP group as compared to the TMD/bruxism-nonDAP group. Specific masticatory disorders including difficulties to open, pain on opening, chewing pain, fatigue on chewing, jaw deviation to opening, and change to a soft diet, were significantly more prevalent in the DAP group as compared to the other reference groups. Our conclusions are based upon a review of the literature and on the results of this study: a. DAP is a relatively well defined stage of internal joint derangement (IJD); and b. compared to other groups, DAP patients as a subgroup are relatively more impaired by their masticatory and other functional disorders. This study provides strong support to other studies and demonstrates that DAP is a well differentiated and severe internal joint disorder. PMID- 12555928 TI - Effects of the antigravitary modification of the myotension of asset (MAGMA) therapy on myogenic cranio-cervical-mandibular dysfunction: a longitudinal surface electromyography analysis. AB - This study was aimed at evaluating the effects of a novel physiotherapy machine called MAGMA (AntiGravitary Modification of the Myotensions of Asset) on postural and masticatory muscles of subjects with myogenic cranio-cervical-mandibular dysfunction (CMD), by using surface electromyography (sEMG). Fifteen subjects, nine males and six females (mean age 27.6 years), with CMD were included in the study. The bilaterally monitored muscles were: masseter, anterior and posterior temporalis, digastric, posterior cervical, sternocleidomastoid, and upper and lower trapezius. All muscles were monitored at rest, with a second record of maximal voluntary clenching (MVC) for both the masseter and anterior temporalis. Patients were subjected to MAGMA therapy for one session/week of 30 min over ten weeks. The surface EMG activity was recorded twice, at the baseline and at the end of the therapy. After MAGMA therapy, the sEMG activity at rest of the monitored muscles was significantly better when compared to the baseline; the only exception was the anterior and posterior temporalis muscles which did not improve. On the contrary, with the MVC, all the monitored muscles (masseter and anterior temporalis) significantly improved their sEMG activity. Although more investigations are needed, these results indicate that the use of such antigravitary therapy can provide a tool for resolving myogenic CMD. PMID- 12555929 TI - Variants of ossification in the stylohyoid chain. AB - The purpose of this study was to evaluate the age-related differences in the incidence, length and topographic location of ossification in the stylohyoid chain. Panoramic radiographs of 420 patients (795 reviewed stylohyoid-chains), subdivided into 4 age groups (20 years and under, 21 to 40 years, 41 to 60 years, older than 60 years) were reviewed and examined for the incidence, length and topographic location of stylohyoidal ossification. 245 (30.8%) out of 795 reviewed stylohyoidal chains showed radiological variabilities (elongation of the styloid process and/or ossification of the stylohyoid ligament). With increasing age, there was an increase in the incidence and length of stylohyoidal ossifications (p<0.01). A significant linear correlation between the length of the stylohyoidal ossifications and age was only found in the young age group (under 20 yrs.; p<0.01). In the young age group (under 20 yrs), there was also a higher prevalence of isolated locations in the superior stylohyoidal segment. With increasing age, there was a pronounced presence of ossifications in the middle and inferior stylohyoid segments and combinations of ossified variabilities. Stylohyoidal ossifications show age-related differences in incidence, length and topographical location. Stylohyoid ossifications gain a different importance in adult patients than in juvenile patients. PMID- 12555930 TI - Changes in urinary bone resorption markers (pyridinoline, deoxypyridinoline) resulting from experimentally-induced osteoarthritis in the temporomandibular joint of rats. AB - The purpose of this study was to quantify the urinary bone resorption markers, pyridinoline (Pyr) and deoxypyridinoline (Dpyr), excreted from experimentally induced osteoarthritis (OA) in the temporomandibular joint (TMJ) of rats. Osteoarthritic lesions were induced by intra-articular injection of collagenase into the right TMJs of 16-week-old male rats. The whole day's urine was collected from each animal one day before the injection and 5, 7, 11 and 14 days after the injection. Urine samples were analyzed by high-perfomance liquid chromatography and fluorescence spectroscopy. Histological changes in condyle were examined by using paraffin sections with toluidine blue staining. Degenerative changes were observed in the articular cartilage of the experimental group on day 7 and day 14 after the injection of collagenase. The concentration of Pyr was remarkably high in the experimental group, and consequently the Pyr to Dpyr ratio was significantly higher (p<0.05) in the experimental group than in the control group from 7-14 days after the injection. These findings suggest that a urinary Pyr/Dpyr ratio would be available for the detection of degenerative changes in condyle relevant to temporomandibular joint osteoarthritis (TMJ OA). PMID- 12555931 TI - Magnetoencephalographic study of the starting point of voluntary swallowing. AB - Clear findings relative to where in the brain the starting point of voluntary swallowing is controlled were obtained in the present magnetoencephalographic study. Namely, the cerebral activity was observed in the cingulate gyrus and supplementary motor area for about 80 ms between 1,000 and 1,500 ms before swallowing in all test subjects. Thus, it is clear that this type of central control mechanism also plays an important role in complicated swallowing movements. PMID- 12555932 TI - Symptom relief after treatment of temporomandibular and cervical spine disorders in patients with Meniere's disease: a three-year follow-up. AB - This study describes the coordinated treatment of temporomandibular disorders (TMD) and cervical spine disorders in patients diagnosed with Meniere's disease. The aim was to follow up treatment outcomes for three years with regular follow up examinations every six months. Of the 31 patients with Meniere's disease who participated in a controlled comparative study on the signs and symptoms of TMD, 24 participated in a subsequent controlled comparative study on the signs and symptoms of cervical spine disorders (CSD). These 24 Meniere's disease patients (ten males and 14 females) agreed to participate in this longitudinal study. At each follow-up, their symptoms were evaluated using self-administered questionnaires and visual analog scales (VAS). The results of the coordinated treatment showed simultaneous decreases in the intensities of vertigo, nonwhirling dizziness, tinnitus, feeling of fullness in the ear, pain in the face and jaws, pain in the neck and shoulders, and headache that were both longitudinal and highly significant. Significant longitudinal reductions in the frequencies of vertigo, nonwhirling dizziness, and headache were also reported by the patients as well as a complete disappearance of pain located in the vertex area. A significant relief of TMD symptoms and a decrease in nervousness was also achieved. The results showed that a coordinated treatment of TMD and CSD in patients with Meniere's disease is an effective therapy for symptoms of this disease. The results also suggested that Meniere's disease has a clear association with TMD and CSD and that these three ailments appeared to be caused by the same stress, nervousness, and muscular tension. PMID- 12555933 TI - Mandibular lengthening by distraction osteogenesis for treatment of obstructive sleep apnea syndrome: a case report. AB - A 31-year-old man who had been diagnosed with obstructive sleep apnea syndrome (OSAS) due to retrognathia was referred to our hospital. Before treatment, the angle of the sella-nasion-point B (SNB) was 67 degrees. The lowest oxygen saturation (SpO2) during sleep was 77%, the longest-running episode of sleep apnea was 128 seconds, and his apnea index (AI) was 29.9. He underwent bilateral horizontal mandibular distraction osteogenesis using intraoral distraction devices. After mandibular distraction, SNB increased to 72 degrees. The lowest SpO2 during sleep increased to 85%, the longest-running episode of sleep apnea decreased to 40 seconds, and his AI decreased to 4.1 (within normal range). No marked recurrence of OSAS was observed even at one year after mandibular distraction. This report suggests that mandibular lengthening by distraction osteogenesis is an effectual method for the treatment of OSAS due to retrognathia. PMID- 12555934 TI - Guidelines for diagnosis and management of disorders involving the temporomandibular joint and related musculoskeletal structures. PMID- 12555936 TI - Individual blood-brain barrier phenylalanine transport in siblings with classical phenylketonuria. AB - Recent studies indicate that individual blood-brain transport characteristics of phenylalanine may lead to different clinical outcomes in phenylketonuria (PKU) patients in spite of comparable dietary control. To check these preliminary data, we investigated four pairs of siblings with classical PKU (and identical genotype) using in vivo nuclear magnetic resonance spectroscopy in the course of an oral phenylalanine load (100 mg/kg body weight). Patients' brain phenylalanine concentrations were different in spite of similar blood levels. Interindividual variations of the apparent transport Michaelis constant, Kt,app, ranged from 0.10 to 0.84 mmol/L. The ratio of the maximal transport velocity, Tmax, over the intracerebral consumption rate, Vmet, varied between 2.61 and 14.0. Siblings with lower values for Kt,app, higher values for Tmax/Vmet, and higher concurrent brain phenylalanine levels showed a lower IQ and a higher degree of cerebral white matter abnormalities. The results indicate that blood-brain barrier transport characteristics and the resultant brain phenylalanine levels are causative factors for the individual clinical outcome in PKU. PMID- 12555935 TI - Short-term dietary interventions in children and adolescents with treated phenylketonuria: effects on neuropsychological outcome of a well-controlled population. AB - This study addressed two questions: is there an effect of dietary interventions that induce relatively small changes in phenylalanine (Phe) concentration on neuropsychological outcome of early- and continuously treated phenylketonuria (PKU) patients, and are there differences in effects for PKU children and adolescents? To answer the first question, the effect of a short-term dietary intervention (1-2 weeks) was compared for patients whose Phe concentrations increased vs those whose Phe concentrations decreased. Controls were tested twice to control for learning effects. To answer the second question, the effect of dietary interventions was examined in younger patients (aged 7-10 years) and older patients (aged 11-14 years). The effect of dietary interventions was determined with three neuropsychological tasks: one requiring sustained attention; a second demanding maintenance in working memory; and a third in which complex operations were performed in working memory. Relatively small fluctuations in Phe concentration were found to influence neuropsychological task performance of PKU patients. Patients whose Phe concentrations decreased at the second assessment showed generally more improvement than controls. Patients whose Phe concentrations had increased showed minimal improvement or deterioration of task performance. The strongest effects were observed when sustained attention and manipulation of working memory content were required. There were some indications of a greater sensitivity of younger patients to fluctuations in Phe concentration. PMID- 12555937 TI - Hartnup disorder: polymorphisms identified in the neutral amino acid transporter SLC1A5. AB - Hartnup disorder is an inborn error of renal and gastrointestinal neutral amino acid transport. The cloning and functional characterization of the 'system B0' neutral amino acid transporter SLC1A5 led to it being proposed as a candidate gene for Hartnup disorder. Linkage analysis performed at 19q13.3, the chromosomal position of SLC1A5, was suggestive of an association with the Hartnup phenotype in some families. However, SLC1A5 was not linked to the Hartnup phenotype in other families. Linkage analysis also excluded an alternative candidate region at 11q13 implicated by a putative mouse model for Hartnup disorder. Sequencing of the coding region of SLC1A5 in Hartnup patients revealed two coding region polymorphisms. These mutations did not alter the predicted amino acid sequence of SLC1A5 and were considered unlikely to play a role in Hartnup disorder. There were no mutations in splice sites flanking each exon. Quantitative RT-PCR of SLC1A5 messenger RNA in affected and unaffected subjects did not support systemic differences in expression as an explanation for Hartnup disorder. In the six unrelated Hartnup pedigrees studied, examination of linkage at 19q13.3, polymorphisms in the coding sequence and quantitation of expression of SLC1A5 did not suffice to explain the defect in neutral amino acid transport. PMID- 12555938 TI - Introduction of a ketogenic diet in young infants. AB - The ketogenic diet is a rational treatment for pyruvate dehydrogenase complex deficiency (McKusick 312170) and GLUT1 deficiency syndrome (McKusick 138140). An increasing number of patients are diagnosed in early infancy, but few data are available on the introduction of a ketogenic diet in this age group. GLUT1 deficiency syndrome was suspected in four infants presenting with seizures and unexplained hypoglycorrhachia. A ketogenic diet was introduced at 6-28 weeks of age. Ketosis was initiated by fasting, monitored by bedside blood glucose and 3 hydroxybutyrate determinations, and was maintained successfully using supplemented carbohydrate-free infant formula and emulgated triglycerides. All patients developed ketosis within 24 h. 3-Hydroxybutyrate concentrations available at the bedside correlated inversely with the base excess. At glucose levels < or = 40 mg/dl patients remained asymptomatic in the presence of ketones. The ketogenic formula was tolerated well, parental compliance was good, and all patients remained seizure-free on the diet. GLUT1 deficiency was confirmed in two patients; the diet was discontinued in the other two patients. In one infant, failure to thrive on medium-chain triglycerides was effectively reversed using long-chain triglycerides. Urine dipstick analyses failed to detect ketosis in another infant. Adverse effects of the diet were limited to renal stones in one patient. The ketogenic diet can be introduced and maintained successfully in young infants using long-chain fat emulsion. Monitoring 3-hydroxybutyrate at the bedside was useful for metabolic control and superior to urine dipstick analysis. Seizure control was effective and adverse effects were limited, but evaluation of the long-term effects of the ketogenic diet in this age group must await ongoing studies. PMID- 12555939 TI - CblE type of homocystinuria due to methionine synthase reductase deficiency: clinical and molecular studies and prenatal diagnosis in two families. AB - The cblE type of homocystinuria is a rare autosomal recessive disorder, which manifests with megaloblastic anaemia and developmental delay in early childhood. This disease is caused by a defect in reductive activation of methionine synthase (MTR). Our study was directed at clinical, biochemical, enzymatic and molecular characterization of two Czech patients with the cblE type of homocystinuria. Case 1 involves a 20-year-old mentally retarded patient who presented with megaloblastic anaemia at 10 weeks of age. She was treated with folates and vitamin B12, and subsequent attempts to cease administration of folates led to recurrence of megaloblastic anaemia. Biochemical features included severe hyperhomocysteinaemia and hypomethioninaemia and in fibroblasts defective formation of methionine from formate, and no complementation with cblE cells. Subsequent molecular analysis of the methionine synthase reductase (MTRR) gene revealed compound heterozygosity for a transition c.1459G>A (G487R) and a 2bp insertion (c.1623-1624insTA). Case 2 involves an 8-year-old girl with nystagmus and developmental delay in whom megaloblastic anaemia was detected at 11 weeks of age. Severe hyperhomocysteinaemia with normal methionine levels was found and enzymatic and complementation studies confirmed the cblE defect. This patient is homozygous for a 140 bp insertion (c.903-904ins140). The insertion is caused by a T>C transition within intron 6 of the MTRR gene, which presumably leads to activation of an exon splicing enhancer. In the families of both patients, enzymatic and mutation analyses were successfully used for prenatal diagnosis. Our study expands the knowledge of the phenotypic and genotypic variability of the cblE type of homocystinuria and supports the concept that this disorder is caused by mutations in the MTRR gene. PMID- 12555940 TI - 2-methyl-3-hydroxybutyryl-CoA dehydrogenase deficiency in a 23-year-old man. AB - 2-Methyl-3-hydroxybutyryl-CoA dehydrogenase (EC 1.1.1.178) deficiency is a recently described defect of isoleucine catabolism. The disorder is characterized by normal early development followed by a progressive loss of mental and motor skills. Deterioration may be rapid or may follow a slower decline with a possible stabilization of the disorder on a low-protein diet and appropriate medication. We report a 23-year-old man with 2-methyl-3-hydroxybutyryl-CoA dehydrogenase deficiency with a very mild clinical course. He had apparently normal early development and remained relatively well until the age of 6 years, when he contracted measles. Following this illness, his motor skills and school progress deteriorated. At 15 years he had significant dysarthria, and generalized rigidity with some dystonic and unusual posturing. He was then treated with a low-protein high-carbohydrate diet with a good response in terms of balance and gait. At 18 years he was given benzhexol (Artane), increased slowly from 2 mg to 6 mg daily, resulting in improvement in tremor and dystonia. At 23 years he can dress himself and works in sheltered employment but remains severely dysarthric. PMID- 12555941 TI - Glutaric aciduria type III: a distinctive non-disease? AB - Glutaric aciduria type III is a rare metabolic abnormality leading to persistent isolated glutaric acid excretion. We report the clinical and biochemical phenotypes of three affected children. The first patient is a boy with dysmorphic features and a chromosomal deletion (monosomy 6q26-qter) in whom a persistent glutaric aciduria (500 mmol/mol creatinine, normal <10) was detected during a routine metabolic investigation. The second boy suffered from acute gastroenteritis and hyperthyroidism, when an excessively high urinary glutaric acid excretion was identified (1460 mmol/mol creatinine). The third patient is a girl with constantly elevated glutaric acid in her urine (290 mmol/mol creatinine) but no symptoms of significant disease. In all our patients, glutaric aciduria type I (glutaryl-CoA dehydrogenase deficiency), glutaric aciduria type II (multiple acyl-CoA dehydrogenation defect), and secondary forms of glutaric aciduria (for example due to intestinal infections or mitochondrial dysfunction) could be excluded. Loading with the precursor amino acid lysine in all patients as well as with pipecolic acid in the third case led to an increase in urinary glutaric acid excretion, proving the endogenous origin of glutarate. Glutaric aciduria type III (a defect reported to be caused by peroxisomal glutaryl-CoA oxidase deficiency) is our presumptive diagnosis. However, peroxisomal glutaryl CoA oxidase is not well characterized and no reliable approach for the direct determination of this enzyme is available to us. To our knowledge, in the English language literature only a single patient with glutaric aciduria type III has been described. Our cases reported here confirm the earlier assumption that glutaric aciduria type III is not related to a distinctive phenotype. Glutaric aciduria type III appears to be a rare metabolic abnormality, presumably of peroxisomal metabolism. However, its pathophysiological impact still needs further investigation. PMID- 12555942 TI - Niemann-Pick disease type C in adults. AB - Although it is often perceived as a paediatric disorder, significant numbers of patients with Niemann-Pick disease type C present for the first time in adult life or survive into adult life. The presentation in these patients differs from that seen in the classical juvenile form of the disease. Adult patients are often referred to clinicians with psychosis or other major psychiatric problems. The dystonia with preserved intellectual functioning can be mistaken for other basal ganglia disorders such as Wilson disease. The presence of vertical gaze palsy is an important clinical clue and, in the presence of a modest increase in plasma chitotriosidase activity, can be very helpful in the differential diagnosis. The diagnosis should be confirmed in suspected cases by filipin staining of cultured fibroblasts, as well as cholesterol esterification studies and DNA mutation analysis. PMID- 12555943 TI - Mutations in the sterol 27-hydroxylase gene (CYP27A) cause hepatitis of infancy as well as cerebrotendinous xanthomatosis. AB - Follow-up investigations were undertaken on a previously reported patient who had severe familial giant cell hepatitis in infancy associated with substantially increased urinary excretion of bile alcohol glucuronides. By the age of 11 years, he had developed a profile of cholanoids in plasma and urine that closely resembled the pattern seen in cerebrotendinous xanthomatosis (CTX). Sequencing of the sterol 27-hydroxylase gene (CYP27A) showed that he was homozygous for a deletion (525/526delG) that causes a frameshift and a premature stop codon. This genotype has previously been described in an adult female with classical symptoms of CTX (tendon xanthomata, cataracts and deteriorating cognitive function). A review of past medical histories of a group of patients with CTX revealed that prolonged neonatal cholestatic jaundice was common. The family histories also revealed fetal and neonatal deaths among siblings of patients with CTX. We conclude that defective activity of cholesterol 27-hydroxylase can lead to neonatal cholestatic jaundice ('hepatitis of infancy'), which may be self limiting. After a latent period, however, progressive accumulation of cholesterol and cholestanol can lead to the xanthomata, neurodegeneration, cataracts and atherosclerosis that are typical of CTX. PMID- 12555944 TI - Ramipril treatment in a patient with glycogen storage disease I non-A. AB - We studied the effect of ramipril on proteinuria and mild hypertension in a 21 year-old patient affected by glycogen storage disease type I non-A. After few months of therapy we obtained a decrease in total urine protein excretion that later re-increased in spite of the high dose of ACE inhibitor. Even if ACE inhibitors are the only effective therapy for GSD I nephropathy, further studies are requested. PMID- 12555945 TI - Prenatal diagnosis of succinate semialdehyde dehydrogenase deficiency in non identical twins. AB - Prenatal diagnosis was performed by both DNA and enzymatic analysis on non identical twins conceived by in vitro fertilization and at risk of succinate semialdehyde dehydrogenase deficiency. One fetus was predicted to be affected and one unaffected and selective fetal reduction was performed. PMID- 12555946 TI - Selective screening of amino acid disorders in the south-west of Iran, Shiraz. AB - A diagnostic study of amino acid concentrations in dried blood spot samples from 1044 symptomatic children revealed high incidences of phenylketonuria, tyrosinaemias, and maple syrup urine disease in the Shiraz region of Iran. Minimum incidences, based on babies born between 1996 and 2001 inclusive, and diagnosed by the end of 2001, are 1:3672, 1:10651 and 1:21 303, respectively. PMID- 12555947 TI - Unknown peaks in GC-MS analyses of Guthrie cards blood samples might be the result of sample contamination by the black markings of the card. AB - Azelaic acid, sometimes found during analysis of Guthrie card blood samples, may be derived from the printing ink used to mark the spotting area. PMID- 12555948 TI - Mutation screening for tyrosinaemia type I. AB - This study reports the development of a mutation screening strategy for tyrosinaemia type I, and the identification of six novel mutations in the FAA gene. PMID- 12555949 TI - A novel mutation in the gene for canine acid beta-galactosidase that causes GM1 gangliosidosis in Shiba dogs. AB - A homozygous recessive mutation, causing GM1-gangliosidosis in Shiba dogs, was identified as a deletion of C nucleotide 1668 in the gene for canine acid beta galactosidase, which was a novel mutation in canine GM1-gangliosidosis. PMID- 12555950 TI - If a tree falls ... getting your clinical insights out. PMID- 12555951 TI - Shade match. PMID- 12555952 TI - Women's health. PMID- 12555953 TI - Employee attitudes. PMID- 12555954 TI - Do you have any interest in relocating to another region of the country? PMID- 12555955 TI - The technologically well-equipped dental office. AB - BACKGROUND: Information technology, or IT, applications for dental practice continue to develop rapidly. More than 85 percent of all dentists use computers in their offices, and the number of clinical uses for the computer is on the rise. This article discusses the state of the art of several technologies, provides an integrative view of a technologically well-equipped office and offers several guidelines for technology purchasing decisions. CONCLUSIONS: Most administrative applications are relatively mature and are required for the smooth functioning of today's dental offices. Clinical applications--such as dental and medical health histories, charting, digital imaging, diagnostic and treatment applications, and decision support--vary in their degree of maturity. Most Internet applications-such as Internet-based scheduling and e-mail communication with patients--still are an optional adjunct for dental practices. PRACTICE IMPLICATIONS: Practitioners should develop a comprehensive plan for implementing or updating the IT infrastructure in their offices. Issues to be considered in technology purchasing decisions include usability, integration, work flow support, cost-benefit analysis and compliance with standards. PMID- 12555956 TI - The effect of diabetes mellitus on endodontic treatment outcome: data from an electronic patient record. AB - BACKGROUND: The authors used a custom-built electronic record system to investigate endodontic diagnostic and treatment outcome data in patients with and without diabetes. METHODS: The medical histories and endodontic treatment data for nonsurgical endodontic patients treated in predoctoral and postgraduate specialty clinics were entered into an electronic record system. A total of 5,494 cases (including 284 cases in patients with diabetes) were treated, and 540 cases (including 73 cases in patients with diabetes) had follow-up data two years or more postoperatively. The authors performed univariate and multivariate analyses to determine important factors affecting endodontic diagnosis and treatment outcome. RESULTS: Patients with diabetes had increased periodontal disease of teeth with endodontic involvment compared with patients who did not have diabetes. There was a trend toward increased symptomatic periradicular disease in patients with diabetes who received insulin, as well as flareups in all patients with diabetes. Two years or longer postoperatively, 68 percent of cases followed were successful. Older age, the absence of preoperative lesions, the presence of permanent restorations and longer postoperative evaluation periods all were associated with a successful outcome. A multivariate analysis showed that in cases with preoperative periradicular lesions, a history of diabetes was associated with a significantly reduced successful outcome. CONCLUSIONS: Patients with diabetes have increased periodontal disease in teeth involved endodontically and have a reduced likelihood of success of endodontic treatment in cases with preoperative periradicular lesions. CLINICAL IMPLICATIONS: Patients with diabetes who are treated endodontically should be assessed carefully and be treated with effective antimicrobial root canal regimens, particularly in cases with preoperative lesions. PMID- 12555957 TI - Using chemomechanically assisted diamond bur cutting for improved efficiency. AB - BACKGROUND: Surface active agents added to handpiece coolants enhance bur cutting rates, or CRs, through chemomechanical effects, or CMEs. The authors evaluate the effect of CMEs on long-term cutting by diamond burs. METHODS: The authors tested medium-grit diamonds in a high-speed handpiece under a 147.5-gram load at 350,000 revolutions per minute, with 22 milliliters per minute coolant flow consisting of distilled water or a 1:10 mouthwash/water mixture. Repeated 6-millimeter-long edge cuts were made through machinable glass ceramic bars until the CR (determined as the time to transect the bars) had decreased by more than 75 percent. The authors used six burs for each coolant and analyzed the data via one way analysis of variance with post hoc Scheffe tests. RESULTS: CRs with water irrigation continuously decreased with the number of cuts and declined by 87 percent over eight cuts. The authors found faster CRs with CMEs; after 12 cuts, the CR was still close to 40 percent of the initial CR, compared with only 13 percent of the initial CR for water irrigation after eight cuts. These differences between CME-enhanced CRs and those found with water irrigation were statistically significant. CONCLUSIONS: Diluted alcohol- and glycerol-based mouthwash/water mixtures significantly enhanced the CRs of diamond burs and prolonged their service life by more than 50 percent compared with water irrigation alone. CLINICAL IMPLICATIONS: Dentists can increase bur CRs and extend bur cutting life by as much as 200 percent through the addition of diluted alcohol and glycerol mouthwash to the handpiece coolant. PMID- 12555958 TI - Xerostomia: etiology, recognition and treatment. AB - BACKGROUND: Clinicians may encounter symptoms of xerostomia, commonly called "dry mouth," among patients who take medications, have certain connective tissue or immunological disorders or have been treated with radiation therapy. When xerostomia is the result of a reduction in salivary flow, significant oral complications can occur. TYPES OF STUDIES REVIEWED: The authors conducted an Index Medicus--generated review of clinical and scientific reports of xerostomia in the dental and medical literature during the past 20 years. The literature pertaining to xerostomia represented the disciplines of oral medicine, pathology, pharmacology, epidemiology, gerodontology, dental oncology, immunology and rheumatology. Additional topics included the physiology of salivary function and the management of xerostomia and its complications. RESULTS: Xerostomia often develops when the amount of saliva that bathes the oral mucous membranes is reduced. However, symptoms may occur without a measurable reduction in salivary gland output. The most frequently reported cause of xerostomia is the use of xerostomic medications. A number of commonly prescribed drugs with a variety of pharmacological activities have been found to produce xerostomia as a side effect. Additionally, xerostomia often is associated with Sjogren's syndrome, a condition that involves dry mouth and dry eyes and that may be accompanied by rheumatoid arthritis or a related connective tissue disease. Xerostomia also is a frequent complication of radiation therapy. CONCLUSIONS AND CLINICAL IMPLICATIONS: Xerostomia is an uncomfortable condition and a common oral complaint for which patients may seek relief from dental practitioners. Complications of xerostomia include dental caries, candidiasis or difficulty with the use of dentures. The clinician needs to identify the possible cause(s) and provide the patient with appropriate treatment. Remedies for xerostomia usually are palliative but may offer some protection from the condition's more significant complications. PMID- 12555959 TI - Antidepressant use in psychiatry and medicine: importance for dental practice. AB - BACKGROUND: Many dental patients receive antidepressant therapy. However, antidepressants taken with other drugs may increase the risk of complications that require special dental precautions and care. METHODS: The authors conducted a retrospective study of 1,800 randomly selected patient records and evaluated the prevalence of using antidepressants and other medications concurrently. They analyzed antidepressant intake relative to drug classification and mechanism of action, age, sex and associated potential for clinical complications such as xerostomia, orthostatic hypotension and interaction with vasoconstrictors. The potential for additive adverse effects between antidepressants and other medications also was analyzed. RESULTS: Three hundred eighty-one (21 percent) of the 1,800 patient records indicated that patients were being treated with 412 antidepressants. Female subjects out-numbered male subjects by an approximate 2.3:1 ratio. Selective serotonin reuptake inhibitors were most commonly prescribed, followed by tricyclic antidepressants, atypical and third-generation antidepressants, and monoamine oxidase inhibitors. Based on reported medication intake, almost 58 percent of subjects in the antidepressant group were receiving treatment with two or more medications that had the potential for producing xerostomia. Two hundred fifty-seven (67 percent) of the 381 records documented intake of an antidepressant or other medication with orthostatic hypotension potential. CONCLUSIONS: Three hundred eighty-one patients reported that they were receiving antidepressant therapy for psychiatric and other medical reasons. Potential adverse effects and interactions with other medications have direct implications for dental treatment. CLINICAL IMPLICATIONS: Patients receiving antidepressant therapy are at risk of developing xerostomia and orthostatic hypotension, as well as experiencing the adverse effects of interaction with vasoconstrictors. Dentists must take appropriate precautions in treating these patients. PMID- 12555960 TI - Branchial cleft or cervical lymphoepithelial cysts: etiology and management. AB - BACKGROUND: The cervical lymphoepithelial or branchial cleft cyst is a developmental cyst that has a disputed pathogenesis. The objective of this article is to provide a brief review of the literature and to define diagnostic terms related to this anomaly, as well as to describe its etiology, clinical presentation and treatment. CASE DESCRIPTION: The cervical lymphoepithelial or branchial cleft cyst usually presents as a unilateral, soft-tissue fluctuant swelling that typically appears in the lateral aspect of the neck, anterior to the sternocleidomastoid muscle, and becomes clinically evident late in childhood or in early adulthood. Clinicians can diagnose the cyst with appropriate imaging to assess the extent of the lesion before definitive surgical treatment. The authors describe a patient who underwent excision of a well-encapsulated cystic structure that was diagnosed as a branchial cleft cyst. CLINICAL IMPLICATIONS: The cervical lymphoepithelial or branchial cleft cyst can be easily misdiagnosed as a parotid swelling or odontogenic infection. It is imperative that clinicians make an accurate diagnosis so that appropriate treatment (that is, surgical excision) can be performed. If the cysts are treated properly, recurrences are rare. PMID- 12555961 TI - Minimally invasive dentistry. AB - BACKGROUND: During the past few decades, scientific developments in cariology, dental materials and diagnostic systems have changed dentistry's approach to diagnosis and management of dental caries. The authors summarize these developments. OVERVIEW: Dental adhesives and restorative materials, new understanding of the caries process and remineralization, and changes in caries prevlance have catalyzed the evolution in caries management from G.V. Black's "extension for prevention" to "minimally invasive." The authors describe the scientific basis for early diagnosis; a modified classification of caries based on site and size of lesion remineralization; reduction of cariogenic bacteria; and minimally invasive cavity preparation design, techniques and material selection. CONCLUSIONS AND PRACTICE IMPLICATIONS: Minimally invasive dentistry is based on advances in science. Emerging technologies will facilitate evolution to primary prevention of caries, though technical, cultural and economic obstacles to full implementation in clinical practice now exist. PMID- 12555962 TI - The effect of external nasal dilators on blood oxygen levels in dental patients. AB - BACKGROUND. The authors conducted a study to examine the use of external nasal dilators, or ENDs, on dental patients and to determine whether the subjective improvement in comfort level noted in dental patients using ENDs is accompanied by a rise in the level of oxygen saturation as measured by pulse oximetry, or SpO2. METHODS: The authors used a hand-held pulse oximeter to monitor 45 patients' SpO2 levels during routine dental procedures. RESULTS: The data suggest that dental procedures in general result in a lowering of the SpO2 level and that patients using an END do experience a rise in the SpO2 level. CONCLUSIONS: The data do not clearly establish that the increase in patient comfort with use of an END is due to a rise in the SpO2 level. CLINICAL IMPLICATIONS: The results of this study demonstrate that the use of an END facilitates nasal breathing in dental patients. PMID- 12555963 TI - ADA members weigh in on critical issues. AB - BACKGROUND: Science, new technology, patient care, dental reimbursement and government regulations all affect today's dental practitioners. To find out more about how such challenges may affect current private practitioners, the American Dental Association conducted the 2000 Membership Needs and Opinions Survey. METHODS: A questionnaire was sent to 6,310 ADA members in January 2000 with follow-up mailings in February, March and April 2000. Data collection was completed in July 2000. The survey included questions on critical professional issues, and on perceptions of the ADA and ADA priorities. A total of 3,558 completed surveys were received for an adjusted response rate of 59.5 percent. RESULTS: Members rated the identified issues' level of importance to them. The top three issues included "maintaining my ability to recommend the treatment option I feel is most appropriate for my patients," "receiving fair reimbursement for the dental services I provide," and "protecting myself, my staff and my patients from communicable diseases." New dentists found other items to be more significant to them compared with members overall. CONCLUSIONS: Although ADA members as a whole had similar views on critical issues facing dentistry and ADA priorities, there were significant differences regarding some issues. New dentists were far more concerned about securing funds for their practice and paying off debt than were all ADA members. Minority dentists expressed greater levels of concern about certain issues than did all ADA members. PRACTICE IMPLICATIONS: When planning and implementing ADA activities, the Association should continue to take into account members' relative rankings of professional issues and note issues of special interest to selected membership subgroups. PMID- 12555964 TI - Oral cleansers. AB - Toothette Disposable Oral Swabs are oral cleansers intended for hospital and nursing home patients who cannot practice normal oral hygiene methods. The foam applicator contains sodium bicarbonate and is grooved to facilitate the removal of thick ropy saliva. The Toothette Single Use System is a kit that includes the Toothette Disposable Oral Swab, tubes of moisturizer and Prox-A-Mint solution, which is an oral debriding agent. The swab can be connected to a suction apparatus, allowing a safe and thorough oral cleaning, which is particularly useful in intubated and non-responsive patients. PMID- 12555965 TI - The importance of oral health in patients receiving long-term care. PMID- 12555966 TI - What has happened to removable partial prosthodontics? PMID- 12555967 TI - Business associate agreements: a key requirement under the HIPAA privacy regulations. Health Insurance Portability and Accountability Act. PMID- 12555968 TI - For the dental patient. What you should know about bad breath. PMID- 12555969 TI - Food animal veterinarian shortage causing growing concern. PMID- 12555970 TI - Practitioners spur university to develop NAVLE preparatory course. PMID- 12555971 TI - Calls for marketing methods to promote veterinary medicine. PMID- 12555972 TI - Manufacturer addresses concerns about FIV vaccine. PMID- 12555973 TI - Failure to increase fees, salaries creates concern. PMID- 12555974 TI - Advocate for serologic testing after vaccination. PMID- 12555975 TI - OCD study receives praise. PMID- 12555976 TI - What is your diagnosis? Severe heartworm disease. PMID- 12555977 TI - Compendium of animal rabies prevention and control, 2003. PMID- 12555978 TI - Hantaviruses: etiologic agents of rare, but potentially life-threatening zoonotic diseases. PMID- 12555979 TI - Survey of antimicrobial susceptibility testing practices of veterinary diagnostic laboratories in the United States. AB - OBJECTIVE: To describe antimicrobial susceptibility testing practices of veterinary diagnostic laboratories in the United States and evaluate the feasibility of collating this information for the purpose of monitoring antimicrobial resistance in bacterial isolates from animals. DESIGN: Cross sectional study. PROCEDURES: A questionnaire was mailed to veterinary diagnostic laboratories throughout the United States to identify those laboratories that conduct susceptibility testing. Nonrespondent laboratories were followed up through telephone contact and additional mailings. Data were gathered regarding methods of susceptibility testing, standardization of methods, data management, and types of isolates tested. RESULTS: Eighty-six of 113 (76%) laboratories responded to the survey, and 64 of the 86 (74%) routinely performed susceptibility testing on bacterial isolates from animals. Thirty-four of the 36 (94%) laboratories accredited by the American Association of Veterinary Laboratory Diagnosticians responded to the survey. Laboratories reported testing > 160,000 bacterial isolates/y. Fifty-one (88%) laboratories reported using the Kirby-Bauer disk diffusion test to evaluate antimicrobial susceptibility; this accounted for 65% of the isolates tested. Most (87%) laboratories used the NCCLS (National Committee for Clinical Laboratory Standards) documents for test interpretation. Seventy-five percent of the laboratories performed susceptibility testing on bacterial isolates only when they were potential pathogens. CONCLUSIONS: The veterinary diagnostic laboratories represent a comprehensive source of data that is not easily accessible in the United States. Variability in testing methods and data storage would present challenges for data aggregation, summary, and interpretation. PMID- 12555980 TI - Effects of storage time and temperature on pH, specific gravity, and crystal formation in urine samples from dogs and cats. AB - OBJECTIVE: To determine effects of storage temperature and time on pH and specific gravity of and number and size of crystals in urine samples from dogs and cats. DESIGN: Randomized complete block design. ANIMALS: 31 dogs and 8 cats. PROCEDURE: Aliquots of each urine sample were analyzed within 60 minutes of collection or after storage at room or refrigeration temperatures (20 vs 6 degrees C [68 vs 43 degrees F]) for 6 or 24 hours. RESULTS: Crystals formed in samples from 11 of 39 (28%) animals. Calcium oxalate (CaOx) crystals formed in vitro in samples from 1 cat and 8 dogs. Magnesium ammonium phosphate (MAP) crystals formed in vitro in samples from 2 dogs. Compared with aliquots stored at room temperature, refrigeration increased the number and size of crystals that formed in vitro; however, the increase in number and size of MAP crystals in stored urine samples was not significant. Increased storage time and decreased storage temperature were associated with a significant increase in number of CaOx crystals formed. Greater numbers of crystals formed in urine aliquots stored for 24 hours than in aliquots stored for 6 hours. Storage time and temperature did not have a significant effect on pH or specific gravity. CONCLUSIONS AND CLINICAL RELEVANCE: Urine samples should be analyzed within 60 minutes of collection to minimize temperature- and time-dependent effects on in vitro crystal formation. Presence of crystals observed in stored samples should be validated by reevaluation of fresh urine. PMID- 12555981 TI - Cryptococcal pyelonephritis in a dog. AB - A 5-year-old castrated male Golden Retriever was evaluated for polyuria, polydipsia, and progressive regurgitation thought to be a result of bacterial pyelonephritis and megaesophagus. Bacteriologic culture of urine failed to yield clinically relevant growth, and results of a urine sediment examination were normal. With time, intention tremors and progressive neurologic dysfunction were also observed. At necropsy, a diagnosis of cryptococcal disease was confirmed histologically and immunohistochemically. Findings in the dog of this report were indicative of nephrogenic diabetes insipidus with polyuria and polydipsia caused by cryptococcal pyelonephritis. Neurologic manifestations of systemic cryptococcus infection included megaesophagus, esophageal hypomotility, and regurgitation attributed to localization of cryptococcal organisms in the brain stem in the region of the dorsal motor nucleus of the vagus nerve. To the authors' knowledge, this is the first report of polyuria secondary to cryptococcal pyelonephritis. PMID- 12555983 TI - Use of a test for proteins induced by vitamin K absence or antagonism in diagnosis of anticoagulant poisoning in dogs: 325 cases (1987-1997). AB - OBJECTIVE: To determine usefulness of the test for proteins induced by vitamin K absence or antagonism (PIVKA) to identify anticoagulant-poisoned dogs, compared with one-stage prothrombin time (OSPT) and activated partial thromboplastin time (APTT) tests. DESIGN: Retrospective study. ANIMALS: 325 dogs. PROCEDURE: Comparisons of results of PIVKA, OSPT, and APTT measurements in dogs with anticoagulant poisoning, hepatic disease, disseminated intravascular coagulation, other blood-related disorders, immune-mediated diseases, or other chronic and acute diseases were performed. Median, quartile, and range values were determined. RESULTS: PIVKA tests with a 150-second critical value had > 98% specificity and > 90% sensitivity for diagnosis of anticoagulant poisoning versus > 99% specificity and > 79% sensitivity with a 300-second critical value. Comparison of PIVKA values among diagnostic groups revealed significant differences between dogs with anticoagulant poisoning and all other groups. CONCLUSIONS AND CLINICAL RELEVANCE: The PIVKA test with a 150-second critical value is diagnostically useful for distinguishing anticoagulant poisoning from other coagulopathies. Severe liver disease can cause false-positive results. Administration of vitamin K1 or early evaluation (within a few hours of ingesting anticoagulant) may cause false-negative results. Dogs with PIVKA test values > 150 seconds and clinical signs of anticoagulant poisoning can confidently be considered to have anticoagulant poisoning because of the high test sensitivity and specificity. PMID- 12555982 TI - Surgical and postoperative complications associated with tibial plateau leveling osteotomy in dogs with cranial cruciate ligament rupture: 397 cases (1998-2001). AB - OBJECTIVE: To identify surgical and postoperative complications of tibial plateau leveling osteotomy (TPLO) in dogs with rupture of the cranial cruciate ligament (CCL) and compare their incidence with those reported in the literature for other commonly performed CCL stabilization procedures. DESIGN: Retrospective study. ANIMALS: 346 dogs undergoing 397 TPLO procedures. PROCEDURE: Medical records of dogs undergoing 563 consecutive TPLO procedures were reviewed. Complications were recorded and assigned to groups on the basis of the period during which the complication was observed. RESULTS: 397 TPLOs met the criteria for inclusion in the study. Complications (n = 136) were recorded in 113 of the 397 (28%) procedures. Multiple complications developed in 10 dogs. In 19 dogs, a second surgery was performed to manage complications. Development of a complication after surgery was not associated with age or body weight of the dog, tibial plateau angle prior to stifle joint surgery, or experience of the surgeon. Factors significantly associated with complications were breed and performance of an arthrotomy concomitantly with TPLO. CONCLUSIONS AND CLINICAL RELEVANCE: TPLO was associated with development of numerous complications, some of which required surgical correction. Most complications resolved with nonsurgical treatment. Several complications were unique to the TPLO procedure because of the surgical technique and implants required. Although TPLO was associated with a greater number of complications than other CCL stabilization methods, the incidence of major complications was similar. PMID- 12555984 TI - Case-control study of late-term abortions associated with mare reproductive loss syndrome in central Kentucky. AB - OBJECTIVE: To identify factors associated with abortions of mares during late gestation attributed to mare reproductive loss syndrome (MRLS). DESIGN: Case control study. ANIMALS: 282 broodmares from 62 farms in central Kentucky, including 137 mares that had late-term abortions (LTAs) associated with MRLS, 98 mares from the same farms that did not abort, and 48 mares that aborted from causes other than MRLS. PROCEDURE: Farm managers were interviewed to obtain data on a wide range of management practices and environmental exposures for the mares. Data for case and control horses were compared to identify risk factors for a mare having a MRLS-associated LTA (MRLS-LTA). RESULTS: Several factors increased the risk of mares having MRLS-LTAs, including increased amount of time at pasture, less time in a stall, feeding concentrate on the ground, higher proportion of diet derived from grazing pasture, being fed in pasture exclusively during the 4-week period prior to abortion, access to pasture after midnight during the 4-week period prior to abortion, and drinking from a water trough or not having access to water buckets or automatic waterers. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis indicates that exposure to pasture predisposed mares to having MRLS-LTAs and stillborn foals. Methods for limiting exposure to pasture (keeping mares in stalls longer) during environmental conditions similar to those seen in 2001 should reduce the risk of mares having MRLS-LTAs. PMID- 12555986 TI - Progress in gastrointestinal pathology in the genetic era. PMID- 12555985 TI - Case-control study of early-term abortions (early fetal losses) associated with mare reproductive loss syndrome in central Kentucky. AB - OBJECTIVE: To identify factors associated with abortions during early gestation classified as mare reproductive loss syndrome (MRLS). DESIGN: Case-control study. ANIMALS: 324 broodmares from 43 farms in central Kentucky, including 121 mares from 25 farms that had early-term abortions (ETAs) associated with MRLS (case horses), 120 mares from the same farms but that did not abort, and 83 mares from 18 farms that were not severely Impacted by MRLS. PROCEDURE: Farm managers were interviewed to obtain data on various management practices and environmental exposures for the mares. Data for case and control horses were compared to identify risk factors for mares having MRLS-associated ETAs. RESULTS: Several factors increased the risk of MRLS-associated ETAs, including feeding hay in pasture, greater than usual amounts of white clover in pastures, more eastern tent caterpillars in pastures, abortion during a previous pregnancy, and sighting deer or elk on the premises. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis indicates that certain characteristics of pastures predisposed mares to MRLS associated ETAs. Methods for limiting exposure to pasture (keeping mares in stalls longer) during environmental conditions similar to those of 2001 (ie, sudden freezing in mid-April following warmer-than-usual springtime temperatures and larger-than-usual numbers of eastern tent caterpillars in and around pastures) should reduce the risk of mares having MRLS-associated ETAs. It was not possible to determine whether exposure to white clover or caterpillars were causal factors for MRLS or were merely indicators of unusual environmental conditions that resulted in exposure of mares to a toxic or infectious agent. PMID- 12555987 TI - Gastric carcinoma distal to the cardia: a review of the epidemiological pathology of the precusors to a preventable cancer. AB - A distinctive gastritis precedes the development of cancer distal to the cardia. Helicobacter pylori infection and the use of pickled foods as substitutes for fresh fruits and vegetables constitute the most important environmental factors that generate this gastritis. This review describes the anatomical changes that characterise the step-by-step evolution of a process that begins in childhood and culminates in invasive cancer in middle and old age. Progression of the gastritis can be followed by measuring the host antibody response to the H. pylori infection and by serum assays that indicate loss of parietal cell mass. Cancer of the distal stomach will disappear if adequate, sanitary housing and year-round fresh vegetables are made available to all economic levels of society. Programmes that offer these reforms must be sustained over several generations, since the anatomical changes that precede gastric cancer are probably not reversible and begin early in life. In the absence of these reforms, death from gastric cancer may be prevented if patients with asymptomatic, early cancers are identified. High H. pylori antibody levels and serum pepsinogen assays may be used to identify persons with the extensive gastritis that favours the presence of such early cancers. PMID- 12555988 TI - Premalignant conditions of the pancreas. AB - Premalignant conditions of the pancreas include benign tumours of the pancreas, intraepithelial neoplasia arising within pancreatic ducts, and tumours of the neuroendocrine cells of the pancreas. In addition, there is a variety of rare genetic conditions that predispose to pancreatic exocrine malignancies such as Peutz-Jeghers syndrome, hereditary non-polyposis colorectal cancer syndrome, familial pancreatitis, germline BRCA2 mutations, and pancreatic endocrine malignancies such as type 1 neurofibromatosis (von Recklinghausen's disease) and multiple endocrine neoplasia type 1. More controversial is the concept of chronic pancreatitis and diabetes mellitus as conditions that increase the risk of pancreatic cancer. However, there is no doubt that smoking is a potentiating factor for pancreatic cancer, especially in people who have familial/genetic risk factors. This review will include the recently proposed new nomenclature and classification system for intraepithelial neoplasia in the pancreatic ducts, an overview of the various familial syndromes that are associated with an increased risk of pancreatic tumours, the surveillance programmes that have been introduced to monitor such families, and methods for early diagnosis. PMID- 12555989 TI - Dysplasia can be a pain in the gut. AB - In the gastrointestinal tract, the term 'dysplasia' is used to refer to non invasive neoplastic epithelium. Although we recognise adenomas of the gut as dysplastic, we don't use the term 'dysplasia' in reporting them. The use of the term 'dysplasia' for pre-invasive epithelium gradually came to replace other terms in studies attempting to identify epithelial changes that were cancer precursors in surveillance biopsies of patients with chronic colitides. The two tier system of classification, dividing dysplasias into low-grade or high-grade lesions is conceptually straightforward; however, difficulties exist in the distinction of regenerative epithelium from low-grade dysplasia, of low-grade from high-grade dysplasia, and in identifying superficially invasive carcinoma in a dysplastic mucosa. The category 'indefinite for dysplasia' is an honest recognition of the difficulties in distinguishing reactive or regenerative epithelium from low-grade dysplasia, since these epithelia share many cytological features. In surveillance biopsies in ulcerative colitis and Barrett's mucosa, for each epithelial category from non-dysplastic through indefinite, low-grade and high-grade dysplasia, there are specific management recommendations that vary from no change in surveillance to increased surveillance to definitive therapy that is often a major resection. The recommendation for referral of high-grade dysplasias to consultants reflects the concern pathologists have about making such clinically significant diagnoses with limited experience. Pathologists should use the accepted terminology, share cases to expand their experience, and seek consultation in selected cases. This paper follows the evolution of the dysplasia concept, details the difficult areas of diagnosis, and discusses the importance of interaction between clinicians and pathologists in dealing with dysplasias. PMID- 12555990 TI - Mutation searching in colorectal cancer studies: experience with a denaturing high-pressure liquid chromatography system for exon-by-exon scanning of tumour suppressor genes. AB - AIMS: In hereditary colorectal cancer (CRC) disorders such as familial adenomatous polyposis and hereditary non-polyposis colon cancer, the identification of germline mutations greatly assists in the clinical management of families. In addition, study of somatic mutations in the cancers themselves (both hereditary and sporadic) has been fundamental in the elucidation of the initiation and progression of CRC. Many of the genes underlying CRC development are large; hence mutation screening is a time-consuming and labour-intensive process requiring a rapid and accurate alternative to gel-based systems such as single-strand confirmational polymorphism (SSCP) or denaturing gradient gel electrophoresis (DGGE). Here we report our progress using denaturing gradient high-pressure liquid chromatography (DHPLC) in the screening of the mismatch repair genes MLH1 and MSH2 and in screening the APC and HPP1 tumour suppressor genes for mutations. METHODS: Genomic DNA was amplified using intronic primer sets spanning individual exons in the gene(s) under study. PCR products were subjected to DHPLC and the resultant chromatographs were compared with those of normal controls and aberrant peaks identified. Amplified products with aberrant peaks in the study samples underwent manual sequencing to confirm the presence of sequence variants. RESULTS: The proportion of amplified fragments showing aberrant peaks (hits) ranged from 18 to 30% and in the case of every gene, more than 80% of these could be confirmed as a sequence variant by manual sequencing. The highest rate was found in HPP1, where all hits were found to be sequence variants, and the lowest rate was found in MSH2, where manual sequencing failed to find a sequence variant in 17% of the hits attained. Mutations varied in their nature from directly truncating through splice variants to missense and deletion mutations. Traces for each mutation displayed unique shapes and both deletions and single base changes were equally dramatic. During the mutation scanning many polymorphisms presented as aberrant peaks, as would be expected. Importantly, the same polymorphism gave an identical chromatographic tracing between individuals, opening the possibility to identify common polymorphisms on pattern recognition alone. There remains, though, the possibility that rare pathogenic variants may assume an identical shape. CONCLUSIONS: The results indicate that DHPLC is a sensitive and efficient technique for screening of DNA for sequence variants. Given that polymorphisms comprised the largest proportion of variants found in each gene (66-100%), excluding these by pattern recognition would markedly reduce the amount of sequencing required. PMID- 12555991 TI - Molecular tumour clocks and colorectal cancer: seeing the unseen. AB - Recent advances in mathematics and sequencing have revolutionised the analysis of evolution. Modern phylogeny is molecular phylogeny, or histories reconstructed from sequences. The same quantitative sequence approaches have not been fully translated to colorectal cancer. Molecular tumour clocks provide opportunities to reconstruct individual tumour histories. Phenotypic and genetic progression are usually thought to be synonymous, but many mutations may accumulate in normal appearing cells. Although such occult genetic progression is essentially invisible, molecular tumour clocks offer the somewhat magical ability to reconstruct what may never be seen. Potentially much of colorectal cancer progression is unseen and unexplored because tumours usually appear late in life. PMID- 12555992 TI - Correlation of mismatch repair genes immunohistochemistry and microsatellite instability status in HNPCC-associated tumours. AB - AIM: The aim of this study was to assess the performance of immunohistochemistry using antibodies for MLH1, MSH2, MSH6 and PMS2 mismatch repair gene proteins against microsatellite instability (MSI) testing. METHODS: Tumour samples included in this study were derived from referred patients for screening for hereditary non-polyposis colorectal cancer (HNPCC) and patients who had resections for colorectal cancer that were examined at our institution. MSI was assessed at nine loci (BAT25, BAT26, BAT40, D2S123, D10S197, D17S579, D18S34, D5S346 and D17S250) in all cases. Immunohistochemistry for MLH1 and MSH2 was performed in all cases. Staining for MSH6 and PMS2 was performed in selected cases only. RESULTS: There were 742 tumours including 661 colorectal lesions and 81 extracolonic tumours of the HNPCC spectrum. Among the 555 MSI-negative tumours, 554 showed an intact protein expression. Amongst the 187 MSI-positive tumours, 126 showed abnormal expression of MLH1 gene protein, 41 showed abnormal expression of MSH2 gene, three showed abnormal expression of MSH6 only, one showed abnormal expression of PMS2 gene protein only and one case showed abnormal expression of all four proteins. CONCLUSION: Immunohistochemistry offers an alternative method for assessment of MSI status which is fast and relatively inexpensive compared with MSI testing. We achieved a sensitivity rate of 92% and specificity of 99.8% for immunohistochemistry testing assessed against the MSI testing. It has to be accepted that a small fraction of MSI-positive cases will be missed by testing with immunohistochemistry alone. PMID- 12555993 TI - The serrated neoplasia pathway. AB - The concept of a 'serrated neoplasia pathway' refers to a pattern of progression of neoplasms of the colon and rectum that involves hyperplastic polyps and serrated adenomas and which results in the development of carcinoma. The existence of this pathway was initially suggested on morphological grounds. Over the past few years, the increasing recognition of biological and genetic similarities in lesions of this pathway has served to reinforce this concept. The likely existence of such a distinct pathway of colorectal carcinogenesis has implications for the practice of surgical pathology. Most notably, it requires pathologists to recognise the entity of the serrated adenoma, and also to recognise those features of hyperplastic polyps that may be associated with a potential for neoplastic progression. PMID- 12555994 TI - Phenotypic expression of gastrointestinal differentiation markers in colorectal adenocarcinomas with liver metastasis. AB - AIM: The purpose of the present study was to clarify the correlation between phenotypic expression of gastrointestinal differentiation markers and colorectal cancer behaviour, particularly invasion and hepatic metastasis. METHODS: Thirty one cases of advanced colorectal adenocarcinoma (CRC) with liver metastasis were selected. Phenotypic patterns were evaluated immunohistochemically by means of antibodies to CD10, MUC2, and human gastric mucin (HGM). RESULTS: The incidence of MUC2 (45.2%) and HGM (16.1%) expression in CRCs with liver metastasis did not differ from non-metastatic CRCs. In contrast, the incidence of CD10 expression was significantly higher in CRCs with liver metastasis (58.1%) than in control CRCs (21.7%). Phenotypic expression in the liver metastasis carcinomas was similar to that of the primary lesions. CONCLUSIONS: The findings suggest that cases of CRC with CD10 expression are at increased risk of liver metastasis. Even if there is no liver metastasis at laparotomy for CRC, careful follow-up is recommended for CRCs with CD10 expression. PMID- 12555995 TI - The pathogenesis of Crohn's disease in the 21st century. AB - Ten years ago Crohn's disease remained a highly complex disorder with an unknown aetiology and a poorly understood pathogenesis. Research in this field had repeatedly confirmed the presence of an activated intestinal immune system but the factors underlying this had not been identified. Since then, the significance of genetic susceptibility has been established and underlined by the discovery of the NOD2 gene. In turn, this has gone on to signal the importance of the innate immune system and the critical relationship between the gut flora and the intestinal mucosa. These factors, together with independent environmental triggers such as cigarette smoking, form the basis of Crohn's disease pathogenesis in the new century. PMID- 12555996 TI - What is diverticular colitis? AB - Diverticular colitis is the term used to describe a particular pattern of active chronic inflammation in the sigmoid colon affected by diverticular disease, namely the occurrence of luminal mucosal inflammation, whether or not there is evidence of inflammation within and/or around the diverticula themselves. The pathogenesis remains uncertain but is almost certainly multifactorial. In some cases mucosal prolapse, faecal stasis and relative mucosal ischaemia have been implicated as important pathogenetic factors, whilst other cases are clearly the result of a mass effect caused by subserosal peridiverticulitis and suppuration. Symptoms and endoscopic findings are diverse. Histologically, the disease may vary from modest inflammatory changes with vascular ectasia, through classical mucosal prolapse changes, to florid active chronic inflammation, closely mimicking chronic inflammatory bowel disease, especially ulcerative colitis. Thus, accurate clinical and endoscopic correlation is vital for the attainment of the correct diagnosis. Diverticular colitis may respond well to treatment similar to that used for chronic inflammatory bowel disease, adding to the similarities of this disease, notably localised to the sigmoid colon, and ulcerative colitis. Indeed, in a few cases described in the literature, diverticular colitis may 'progress' to otherwise classical ulcerative colitis, suggesting, in some cases at least, a similar pathogenesis. PMID- 12555997 TI - Prognostic factors in anal carcinoma. AB - The pathologist's role in the evaluation of patients with anal squamous carcinoma is now largely restricted to establishing the diagnosis on small biopsies. Staging is performed by imaging techniques and grading is uncertain as the biopsy may not be representative of the whole tumour. Histological subtyping is unreliable and has not been proven to be of prognostic significance. A considerable effort resulting in nearly 50 reports on cytogenetic, flow cytometric, immunohistochemical and other investigations has given new insight into pathogenesis but little guidance with regard to the individual prognosis or choice of treatment. It is concluded that future research should concentrate on larger, probably multicentre series using standardised methods and criteria for evaluation. PMID- 12555998 TI - Test and teach. Incidentally discovered splenic vascular lesion. Littoral cell angioma of the spleen. PMID- 12555999 TI - Autopsies on bodies without antemortem risk factors for HCV, HBV and HIV infections: are they safe? PMID- 12556000 TI - Primary melanotic psammomatous schwannoma of the trachea. PMID- 12556001 TI - Neonatal coagulase-negative staphylococcal meningitis: a report of two cases. PMID- 12556002 TI - Coagulopathy from tiger snake envenoming and its treatment. PMID- 12556003 TI - Pagetoid spread of melanocytes in Spitz naevi. PMID- 12556004 TI - Cost-effectiveness analysis of inhaled corticosteroids in asthma: a review of the analytical standards. AB - PURPOSE: To determine whether published cost-effectiveness studies on inhaled corticosteroids (ICS) in asthma adhered to basic analytical standards as defined in health economic textbooks and in guidelines assessing and comparing efficacy and safety. METHODS: Original cost-effectiveness studies published between 1990 and 2000 in general medical or economic journals were reviewed to assess the adherence to five fundamental methodological principles: (1) design of the study, (2) choice of perspective and corresponding costs, (3) choice of outcome measure, (4) marginal cost analysis, and (5) sensitivity analysis and discussion about external validity. For each principle, the studies were ranked as high, medium or low quality. RESULTS: Most of the 18 studies included were ranked medium on the first two principles. The studies adhered to a higher degree to the remaining three principles. Only three studies were high ranked in all five principles. The number of principles fulfilled increased over time. Studies comparing pharmaceutical products from competing companies were typically short-term studies, designed for other purposes than health economic analyses, and, in general, did not use therapeutically equivalent dosing. CONCLUSIONS: Attention should be drawn to the study design, the weak correspondence between perspective and costs, and especially to the impact of bias in health economic results when comparing different doses of ICSs. PMID- 12556005 TI - Improvement of asthma control with a breath-actuated pressurised metred dose inhaler (BAI): a prescribing claims study of 5556 patients using a traditional pressurised metred dose inhaler (MDI) or a breath-actuated device. AB - A relationship has been reported between sub-optimal inhaler technique and control of asthma symptoms. Randomised controlled trials and systematic reviews may fail to accurately represent this relationship, by excluding patients who are unable to correctly use the reviewed devices. Breath actuated inhalers (BAIs) are reported to be easier to use than metered dose inhalers (MDIs). This study uses a large primary care medical record database (DIN-LINK) to examine the 'real-life' clinical effectiveness of a BAI vs. traditional MDIs. A comparison is made between the quantity of asthma-related medication (beta2 agonist, oral steroids and antibiotics) and healthcare resource use required by children (0-12 years) and adults (13+ years) using either the BAI or an MDI over a 12-month period. Out of 5556 new asthma patients who met the inclusion criteria, 306 used a BAI and 5250 used a traditional MDI. Children and adults using the BAI received fewer prescriptions for all three medication types than those using a traditional MDI (children: beta2 agonists -18% P=0.036, oral steroids -88% P<0.05, antibiotics 68% P<0.05 and adults: beta2 agonists -10.9% P=0.179, oral steroids -51.2% P<0.05, antibiotics -19.5% P=0.276). All prescription results reached statistical significance, with the exception of beta2 agonist and antibiotic prescription numbers for adult patients. Children and adults using the BAI required less GP consultations for asthma (children -30.3%, P<0.05 and adults -22.9%, P<0.05) and less GP consultations for respiratory infections than those using a traditional MDI (children -35.2%, P=0.001 and adults -10.4%, P=0.236). Adults using the BAI required less outpatient appointments than adults using an MDI (-43.7%, P=0.166). All non-prescription resource results reached statistical significance with the exception of GP consultations for respiratory infections and outpatient visits in the adult group. In conclusion, children and adults using the BAI appear to have better asthma control than children and adults using an MDI as evidenced by fewer relief medication prescriptions and less healthcare resource use. PMID- 12556007 TI - Yield of bronchoscopy in the diagnosis of neoplasm metastatic to lung. AB - The yield of fiberoptic bronchoscopy (FB) in the diagnosis of pulmonary metastases has not been conveniently analyzed. With the advances in the surgery of pulmonary metastases, there is a need to evaluate the diagnostic yield and the usefulness of FB to exclude other diseases with similar radiological patterns. To determine the value of FB in the diagnosis of pulmonary metastases we have retrospectively analyzed our experience in 113 patients with proven pulmonary metastases. An endobronchial lesion was identified in 57/113 (50.4%). The most frequent tumors with endobronchial lesions were thyroid (100%), head-neck (67%) and breast carcinomas (59%). The highest diagnostic yield was obtained combining techniques of brushing, washing and biopsy (72.6%); in cases with endobronchial lesions (84.2%) and with certain histological types (head-neck 100%; breast 90.9% and colon 84.6%). The most frequent radiological findings were single or multiple nodules (77.9%). Atelectasis were associated with endobronchial lesions. In conclusion, bronchoscopy is a valuable diagnostic procedure in selected patients with metastatic lung disease. PMID- 12556006 TI - Cost-effectiveness of salmeterol in patients with chronic obstructive pulmonary disease: an economic evaluation. AB - Chronic obstructive pulmonary disease (COPD) is associated with a large economic and social burden. Few economic studies have examined the benefits of inhaled therapy for patients with COPD. This 16-week study examined the cost effectiveness of salmeterol in this patient group. Patients with a history of COPD were randomised to treatment with salmetrol 50 mcg (229 patients) or placebo (227 patients) twice daily administered by metered-dose inhaler in addition to normal therapy. Forced expiratory volume (FEV1) was measured and patients recorded the impact of their symptoms in a daily diary. Health status was assessed using the St Georges Respiratory Questionnaire. A significantly higher proportion of salmeterol-treated patients had an increase of >15% in FEV1. Salmeterol patients had a higher mean proportion of symptom-free days and nights than those in the placebo group. More salmeterol patients had a clinically significant improvement in health status. Total healthcare costs were increased in the salmeterol group, but hospital and GP visit costs and concurrent COPD medication costs were lower. The reduction in hospital costs was sufficient to offset a substantial portion of the acquisition cost of salmeterol. Addition of salmeterol to COPD patients' current therapy improved lung function, health status at the expense of a modest increase in costs compared with usual therapy. PMID- 12556008 TI - Discriminant analysis in allergic rhinitis and asthma: methacholine dose-response slope allows a good differentiation between mild asthma and rhinitis. AB - Asthma and rhinitis frequently coexist in allergic patients, but nasal symptoms may predominate, leading to asthma underdiagnosis and undertreatment. Discriminant analysis obtains the best differentiation between groups using one or one set of variables. Our aim was to identify the laboratory test [allergen exposure, total and specific serum IgE, lung function, blood eosinophils and, bronchial response and sensitivity to methacholine (Mth) and allergen] or combination of them that allowed the best differentiation between mild asthma and allergic rhinitis. A cross-sectional analysis was performed in 86 Dermatophogoides pteronyssinus allergic rhinitis patients, who were classified according to clinical data as rhinitis plus mild asthma (n = 62) or "pure" rhinitis (n = 24). Bronchial symptoms had been exhaustively evaluated during a 2 years pre-inclusion period. Patients underwent skin tests and bronchial challenge with Mth and allergen. The exposure to D. pteronyssinus allergen (Der pl) was quantified in dust samples. Dose-response curves with Mth [until the FEV1 fell by 40% or the maximal dose (200 mg/ml) was inhaled] were attained. We developed multiple models of discriminant analysis in order to evaluate the capacity of the above variables to differentiate groups. Asthma patients had higher total and specific IgE levels and a greater sensitivity (PD20 values) and response [dose response slope (DRS)] to both Mth and allergen. The model entering these variables was the one that correctly classified more patients (79.2%). The discriminative power of the model that only included Mth-DRS values was similar to the above (78.8%). Bronchial response to Mth is quantitatively different in allergic rhinitis patients who display mild asthma symptoms when compared to those that only report rhinitis, suggesting a distinct bronchial intrinsic behavior. The utilization of complete dose-response curves with Mth allows a good separation between mild asthma and "pure" rhinitis patients and might be useful in the diagnosis of mild asthma. Whether the early detection and treatment of these patients prevents the development of symptomatic asthma needs further evaluation. PMID- 12556009 TI - Systematic assessment of clinical practice guidelines for the management of chronic obstructive pulmonary disease. AB - OBJECTIVE: To systematically evaluate the quality of the development of guidelines for the management of chronic obstructive pulmonary disease (COPD). METHODOLOGY: MEDLINE and Excerpta Medica search for published guidelines, followed by independent evaluation by two reviewers, according to previously reported guideline development quality criteria, on a three-point scale. RESULTS: Five national COPD guidelines and two international COPD guidelines were retrieved. Reviewers demonstrated good inter-observer agreement in assessing the 10 combined guideline development criteria for the seven guidelines [kappa = 0.66]. Guidelines were only partly multi-disciplinary with little or no consumer input, were up to 48 pages in length, and often lacked practical summaries or management flow charts which could have facilitated retrieval of key management recommendations. Almost all the papers were based upon a consensus approach, rather than evidence based, and methods of resolution of differences of opinion were not stated. Patient outcomes, ethical and medico-legal implications were not addressed and six of the guidelines were sponsored directly or indirectly by a single drug company. CONCLUSIONS: In spite of COPD guidelines being reported by major national bodies for over a decade now, most fail to meet important criteria for high-quality guideline development, and evaluation of clinical impact remains undetermined. PMID- 12556010 TI - Effects by 8-bromo-cyclicAMP on basal and organic dust-induced release of interleukin-6 and interleukin-8 in A549 human airway epithelial cells. AB - Inhalation of organic dust from a swine-confinement building leads to an intense inflammatory reaction with an increased number of inflammatory cells and mediators in the upper and lower respiratory tract of previously unexposed subjects. In vitro the dust induces cytokine release from epithelial cells and alveolar macrophages. It is known that intracellular cyclic AMP (cAMP) contributes to the regulation of inflammatory responses. We therefore investigated whether 8-Bromo-cAMP, a cell membrane-permeable cAMP analogue, would influence release of the cytokines interleukin-6 (IL-6) and IL-8 in a human airway epithelial cell line, A549, exposed to a suspension of the organic dust, and to a supernatant prepared by centrifugation (at low g-force) of a suspension of dust. The large particulate matter was thus sedimented, leaving bacteria, whole and cell wall constituents in the supernatant. Cytokine release was measured with enzyme-linked immunosorbent assay (ELISA). The cytokine release induced by a supernatant was 23% (IL-6) and 27% (IL-8) of the release induced by a dust suspension. 8-Bromo-cAMP (1 mM) doubled basal IL-6 release and IL-6 release induced by a dust supernatant (P<0.01), and increased IL-6 release induced by a dust suspension by 19% (P<0.05). 8-Bromo-cAMP did not affect basal IL-8 release, partially inhibited (28%) the release of IL-8 induced by a dust suspension (P<0.01), but increased IL-8 release induced by a dust supernatant by 13% (P<0.05). In summary, expression of the cytokines IL-6 and IL-8 is differentially regulated by 8-Bromo-cAMP, both with regard to basal and dust induced release. The results indicate that 8-Bromo-cAMP attenuated IL-8 release by affecting signaling transductions induced by the particulate fraction. PMID- 12556011 TI - G-CSF application in patients with severe bacterial pneumonia increases IL-10 expression in neutrophils. AB - In severe pneumonia, the application of granulocyte-colony stimulating factor (G CSF) was associated with reduced complications possibly by an induction of anti inflammatory cytokines. It is not clear, whether G-CSF induces interleukin-10 (IL 10) synthesis in neutrophils. In a randomized study, 15 patients with severe community acquired pneumonia were treated either by a single dose of G-CSF and antibiotic therapy (n=8) or antibiotics alone (n=7). Messenger ribonucleic acid (mRNA) expression of IL-10 and tumor necrosis factor alpha of peripheral blood leukocytes was measured using in-situ hybridization (ISH) and reverse transcription-polymerase-chain-reaction (RT-PCR). In addition, the cytokine release of lipopolysaccharide (LPS)-stimulated whole blood was measured by ELISA. We detected increased IL-10 mRNA by ISH (140 +/- 8% vs. -11 +/- 5%, P<0.01) and RT-PCR (126 +/- 16% vs. -28 +/- 3%, P<0.01) in the G-CSF-treated group only. In contrast, LPS-stimulated whole blood cells in vitro released significantly less IL-10 compared to the control group (-38.2 +/- 97 vs. -14.8 +/- 6 pg/ml, P<0.02). There was no significant effect on IL-10 serum protein levels and the TNF-alpha release and expression. IL-10 mRNA was detected predominantly in cluster designation 66b (CD66b) positive nucleated blood cells indicating that polymorphonuclear leukocytes are the main source of IL-10 expression after G-CSF stimulation. G-CSF induces transcription of IL-10 mRNA in neutrophils without increased release. This may be due to posttranscriptional effects. PMID- 12556012 TI - Down-regulation of aquaporin 3 in bronchiectatic airways in vivo. AB - Bronchiectasis is characterized pathologically by permanent abnormal bronchial dilation, and clinically by chronic sputum production. Aquaporin 3 (AQP3), a recently described water channel that is also found in large airway cell membrane, could play a role in the pathogenesis and particularly that of bronchorrhea in bronchiectasis. However, little is known of its in vivo distribution and physiological role in human airways. We have, therefore, performed this quantitative immunohistochemistry study on endobronchial biopsies to evaluate the expression and clinical relevance of AQP3 in patients with idiopathic bronchiectasis (n = 25, 15 F, 64.3 +/- 11.5 years) and control subjects (n = 14, 5 F, 57.5 +/- 12.0 years). Quantitative image analysis was performed to evaluate the expression of AQP3 in the bronchial epithelial cells. Our results show that AQP3 was predominantly expressed in the basal cells of the epithelial layer in both groups. Expression of AQP3 was significantly reduced in the basal, but not columnar, epithelial cells in bronchiectasis compared with control airways (p = 0.02, 0.35). Only bronchiectatic patients with regular sputum production, but not their counterparts, had significant downregulation of epithelial AQP3 expression compared with control airways (p = 0.004, 0.24). Our findings suggest that AQP3 could have an important role in the pathogenesis of increased mucus production in bronchiectasis. PMID- 12556013 TI - Management of multiple drug-resistant tuberculosis. AB - There has been a worldwide increase in multiple drug-resistant tuberculosis (MDR TB) which has in the past been associated with a poor prognosis. In the U.K., about half of the cases live in the London area and we have set out to obtain further information on their treatment and outcome. We examined the risk factors, drug resistance, drug treatment, sputum conversion, and outcome in patients with MDR-TB at three hospitals in South London and diagnosed during the period June 1995-January 1999. Human Immunodeficiency Virus (HIV)-positive patients were excluded. There were 760 patients resident in Lambeth, Southwark and Lewisham Health Authority (LSLHA) who were notified as tuberculosis (TB) during the time period and who were of negative or unknown HIV status. (The population of LSLHA is approx.750,000.) There was a total of 13 patients with MDR-TB, known or presumed to be HlV negative. Their median age was 28 years (range 15-53); nine (69%) were born outside the U.K. and 11 had pulmonary disease; they had organisms resistant to a median of two first-line drugs (range 2-4) and to a median of four of all drugs tested (range 2-10). They received treatment with a median of six drugs (range 3-9). Eight were followed up for at least 3 years (range 3-6) after the completion of treatment; at their last assessment none had features of active TB and all were sputum negative (smear and culture). Two returned to their countries of origin during treatment; they were sputum negative at that time. Two patients are well and continue on treatment in the U.K. One patient (known HIV negative) died following treatment failure. In conclusion, we obtained disease free survival in eight cases of MDR-TB, known or presumed to be HIV negative and followed up for 3 years or more. The prognosis for patients treated at specialised centres is good (and better than is generally believed). We describe a new protocol for the detection and management of MDR-TB. PMID- 12556014 TI - Bronchodilating effects of cumulative doses of formoterol from a novel multi-dose inhaler (Airmax). AB - The bronchodilating properties of formoterol from a novel multi-dose inhaler, Airmax 6 microg and from a single-dose dry powder inhaler Foradil Aeroliser 12 microg were investigated in 31 adult asthmatics with FEV1 > or = 60% predicted and a reversibility > or = 12%. Patients received on a single day four doses of formoterol: cumulative dose 6, 12, 24 and 48 microg from Airmax, or 12, 24, 48 and 96 microg from Aeroliser. The mean FEV1 (SD) from baseline to 1 h after the final cumulative dose increased by 0.81 l from 2.62 (0.58) to 3.43 l (0.70) with Airmax and by 0.85 l from 2.65 (0.60) to 3.5 l (0.68) with Aeroliser. All 90% CIs for all four dose comparisons were within the equivalence range +/- 0.11. There was a higher incidence of hypokalaemia and hyperglycaemia at highest doses during treatment with the Aeroliser than with Airmax. In conclusion, formoterol delivered from Airmax provides a dose-dependent bronchodilating effect which is similar to that obtained by Aeroliser, at double the dose. PMID- 12556015 TI - Increased prevalence of irritable bowel syndrome in patients with bronchial asthma. AB - Irritable bowel syndrome (IBS) is one of the most common diseases of the gastrointestinal tract. IBS may represent a primary disorder of gastrointestinal motility accompanied with motor dysfunction in various extraintestinal sites. Recent studies suggest that IBS is associated with bronchial hyper-responsiveness and bronchial asthma might be more prevalent in IBS patients than in control subjects. The aim of our study was to assess the prevalence of IBS in a cohort of asthmatic patients. We evaluated 150 patients with bronchial asthma (71 males and 79 females, aged 45.1+/-14.9 years) and two control groups including 130 patients with other pulmonary disorder and 120 healthy subjects. All subjects enrolled (asthmatic and controls) completed the Greek version of the Bowel Disease Questionnaire (BDQ). BDQ is a, previously validated, self-report instrument to measure gastrointestinal symptoms. Diagnosis of IBS was based on Rome II criteria. The IBS prevalence was significantly higher in asthmatics (62/150, 41.3%) than in subjects with other pulmonary disorders (29/130, 22.3%, P<0.001) and healthy ones (25/120, 20.8%, P<0.001). For all subjects studied, the prevalence of IBS was significantly higher in females (78/214, 36.4%) than in males (38/186, 20.4%, P<0.001). The IBS prevalence in asthmatic males was 29.5% vs. 15.2% in male patients with other pulmonary disorders (P=0.002) and 14.2% in male healthy subjects (P=0.002). The IBS prevalence in asthmatic females was 51.8% vs. 28.1% in females patients with other pulmonary disorders (P<0.001) and 26.5% in females healthy subjects (P<0.001). None of the asthma medications were associated with increased or decreased likelihood of IBS. We conclude that patients with bronchial asthma have an increased prevalence of IBS. Further studies are needed to clarify the potential pathogenetic mechanisms underlying the association between IBS and asthma. PMID- 12556016 TI - Proliferation of type II pneumocytes in the lung biopsy specimens reflecting alveolar damage. AB - Proliferation of type II pneumocytes may be the most sensitive pathological indicator of alveolitis, and the density of type II pneumocytes reflecting the severity of the recent alveolar damage. Twenty-nine patients were divided into three groups by the severity of ground-glass opacities on the CT scans; the severe (acute), the moderate (subacute) and the mild (chronic) groups. We compared the density of type II pneumocytes in the transbronchial lung biopsy specimens and cell analysis of bronchoalveolar lavage (BAL) fluid with the ground glass opacities. Clinical and laboratory findings and BAL fluid analysis also corresponded well with this grouping. Type II pneumocytes were selectively stained with an antibody against Thomsen-Friedenreich and the density of the type II pneumocytes was expressed as the number per 1 mm alveolar septal length. The densities of the type II pneumocytes in the severe, the moderate, the mild, and the control groups were 21.2+/-1.1, 14.4+/-7.2, 11.0+/-3.3 and 7.5+/-0.9/mm, respectively. There were significant differences between the acute group and the other three groups, and between the control and chronic groups. The density of type II pneumocytes is a useful index for evaluating alveolar damage even in mild alveolitis. PMID- 12556017 TI - Use of a paper disposable cup as a spacer is effective for the first-aid management of asthma. AB - OBJECTIVE: There are many settings in which a spacer device is not available for the administration of bronchodilator. Therefore, we tested whether a paper disposable cup is as effective as a commercially produced spacer to administer bronchodilator. METHODOLOGY: Randomised controlled trial. 50 subjects aged 16-50 years who had wheeze and a greater than 10% decrease in FEV1 after histamine inhalation test (HIT). Subjects were randomised to either the 150 ml paper disposable cup group (CUP) or the commercially produced spacer group (SPACER). Twenty minutes after 400 microg salbutamol was administered FEV1 was measured. The recovery index measured post-bronchodilator FEV1 as a percentage of baseline FEV1. Also, analysis of covariance tested whether recovery of FEV1 was related to the magnitude of the fall following the HIT. RESULTS: There were no statistically significant differences between CUP and SPACER groups in any characteristics. There was no difference for the recovery index (t48=1.14, P=0.26). Regression analyses showed that the relation between the magnitude of the fall in FEV1 during the HIT and the percent recovery was not different between the CUP and SPACER groups (t=-1.2, P<0.23). CONCLUSIONS: A paper disposable cup was effective for the reversal of mild to moderate bronchoconstriction. Therefore, a paper disposable cup can be used for the first-aid management of asthma when there is concern about cross-infection and a commercially produced spacer is not available. PMID- 12556019 TI - An introduction to systematic reviews in respiratory medicine. AB - Reviews are an essential component of modern medicine. The volume of literature is large, even about a single treatment for pulmonary disease. The task of retrieving all the relevant papers, then assessing the evidence to reach a valid conclusion is very time consuming. At every stage there is the risk of sampling error (failure to get all the evidence) and bias (a systematic distortion of the results due to a weakness in the methodology). There are essentially two types of review: narrative reviews that follow no rules, exposing them to sampling error and bias; and systematic reviews that attempt to minimise these effects by following an explicit structure for retrieving all of the evidence and attempting an objective synthesis of the results from the different trials. A good review can serve a number of purposes including: assembling all the relevant evidence in one place, providing a valid estimate of the overall effect of treatment, producing guidance for clinical practice and generating hypotheses for further trials about patients or settings in which the treatment effect may be less or more effective. PMID- 12556018 TI - Expression of the glucocorticoid receptor alpha and beta isoforms in human nasal mucosa and polyp epithelial cells. AB - The lower sensitivity of the inflamed nasal mucosa to glucocorticoids might be related to an increased expression of the glucocorticoid receptor (GR) beta isoform. We investigated GRalpha and GRbeta mRNA expression in epithelial cells from nasal mucosa and nasal polyps. GRalpha mRNA was at least 1000 times more expressed than GRbeta mRNA in both tissues. GRbeta expression (mean+/-SEM of 10(3) cDNA copies/microg of total RNA) was higher in nasal polyps (1.15+/-0.19; n=27; P<0.01) than in nasal mucosa (0.62+/-0.10; n=32). Nasal polyps with > 3% of inflammatory cells had higher GRbeta levels (1.40+/-0.29; n=16) than both nasal mucosa (P<0.01) and polyps with < or = 3% of inflammatory cells (0.80+/-0.18; n=11; P<0.05). No difference in GRbeta expression was found between nasal mucosa and polyps with < or = 3% of inflammatory cells. GRbeta expression correlated with the inflammatory cell number, especially with mast cells (r=0.50, P<0.0001). There was no difference in GRalpha mRNA expression between nasal mucosa and nasal polyps. In summary, GRalpha is far more expressed than GRbeta in both tissues. The increased expression of GRbeta may be related to the presence of inflammatory cells. PMID- 12556020 TI - Are asthma sufferers at risk when consulting chiropractors over the Internet? PMID- 12556021 TI - On values and democratic policy making: the deceptively fragile consensus around market-oriented medical care. AB - Market-oriented strategies, embodied in managed competition, have become the primary focus of contemporary U.S. health policy. This dominance reflects the emergence of a bipartisan coalition of support among political elites. This study traces the historical evolution of elite support for the market and suggests that the consensus favoring managed competition is deceptively fragile, with support riven by cleavages in the values used to judge fairness in the allocation of medical care. A unique data set of matched questions asked of both policy elites and the general public is used to document these differences in ethical norms. The implications of these cleavages help to explain three puzzling aspects of contemporary U.S. health policy: (1) the persisting inability to translate the principles of managed competition into politically feasible reforms, (2) the repeated failures to implement demonstration projects intended to test competitive pricing within the Medicare program, and (3) the inability of state regulations to assuage the public's concerns about managed care. Some prescriptions for a more revealing and effective treatment of market reforms in health policy conclude this study. PMID- 12556022 TI - Any-Willing-Provider laws: their financial effect on HMOs. AB - Any-Willing-Provider (AWP) legislation requires that health plans accept any health care provider who agrees to conform to the plan's conditions, terms, and reimbursement rates. Many states have adopted such legislation, raising questions about its effect on the managed care market. Those favoring this legislation argue that it will reduce restrictions on choice of provider, while opponents argue that it will reduce competition by increasing administrative and medical costs for managed care plans. Using cross-sectional time-series data for the period 1992-1995 (the period during which many of these laws were enacted), this study investigates the effect that these laws have on HMO financial performance. Our results show that "all-provider" AWP laws have a very limited effect on the financial performance measures we examine. "Pharmacy" AWP laws have a more significant effect, but neither type of law appears to affect the overall profitability of HMOs. PMID- 12556023 TI - Impeding Earl Warren: California's health insurance plan that wasn't and what might have been. AB - It is widely believed that the turning point for U.S. health insurance came in 1949 when Congress failed to adopt President Harry Truman's proposal for a national system. The possibility that a system of state-level health plans might have emerged before Truman's plan has received little attention. Yet several attempts to enact such a plan were made in California by Governor Earl Warren in the mid-1940s. Had Warren succeeded, the California example might have been emulated by other states and the United States might have evolved a system similar to Canada's provincial programs. PMID- 12556024 TI - Responding to the immunoglobulin shortage: a case study. AB - In fall 1997, a shortage of intravenous immunoglobulin (IVIG) developed in the United States because of increased demand for the product, reduced supply, and product recalls. This shortage is a useful model for understanding how our health care system responds to scarcity. Although the U.S. government took steps to inform the medical community of the shortage, with few exceptions, the government did not respond to the shortage in a timely or effective manner. Instead, it took a relatively passive role, leaving IVIG manufacturers and distributors, health care institutions, and clinicians to fend for themselves. The shortage likely had an uneven impact on patients, based on the relative market strength of the health care institutions in which they received care and the individual patient's ability to absorb the increasing out-of-pocket costs of scarce IVIG. Market mechanisms have now largely alleviated the shortage and significantly reduced its detrimental impact on patients. However, future shortages of IVIG or other scarce medical products, such as vaccines and antibiotics, would benefit from more immediate and coordinated efforts not only to make sure that scarce health care resources are distributed in a just manner but also to identify and remedy the sources of health product supply problems. PMID- 12556025 TI - Health manpower planning in Slovenia: a policy analysis of the changes in roles of stakeholders and methodologies. AB - A heightened awareness about medical manpower issues can be observed in countries that are in a state of political, economic, and social transition. Slovenia entered the transition process in 1989 and became an independent country in 1991. Transition and independence influenced its health care in several ways. It changed the health care system and its financing (by introducing a Bismarckian style of social insurance). It then redistributed power from the Ministry of Health to several stakeholders. A major change occurred in the labor market in health care when the flow of health professionals from the newly independent countries greatly decreased. The decrease was partly due to the consequences of the war in the Balkans and partly due to independent labor legislation in Slovenia. Transitional changes brought new stakeholders to the scene, with a resulting redistribution of responsibilities for health manpower policies and the use of various methodologies. This policy analysis offers a detailed description of the contextual framework, quantitative data on medical manpower development, and, most important, interviews with representatives of the key stakeholders and study of relevant policy documents. We conclude that all stakeholders underpin the need for a structured approach toward health manpower planning in the form of a more coherent system of planning, decision making, and control. A compromise on mutual responsibilities between the less dominant Ministry of Health and the two new powerful stakeholders, the Health Insurance Institute of Slovenia and the Medical Chamber of Slovenia, seems necessary. PMID- 12556027 TI - Laboratory studies on waste water sludge application for acid soil remediation. AB - In this work the effects of sludge disposal on pH values, buffer capacity and adsorption capacity of an acid soil were studied. A stabilized waste water sludge and a paper mill sludge were employed; the pH values of both sludges were higher than 8. The observed differences between soil-urban sludge and soil-paper mill sludge systems can be ascribed to the nature of the compounds present in the sludges and adsorbed on the soil. Both kinds of sludge are able to modify the natural pH of the soil, the buffer capacity and its capacity to retain metal. These modifications increase with increasing soil-sludge contact time and are higher for paper mill sludges. Temperature affects only the modifications due to urban sludge treatment. PMID- 12556026 TI - Evaluating community partnerships: a reply to Spitz and Ritter. PMID- 12556028 TI - Determination of boron in silicon-doped gallium arsenide by electrothermal atomic absorption spectrometry and ultraviolet-visible spectrophotometry. AB - Two methods have been developed for the determination of boron impurities in silicon-doped gallium arsenide (GaAs) for electronics. The first method employs the electrothermal atomic absorption spectrometry (ETAAS), the second, the UV-Vis molecular absorption spectrophotomety. In both cases the GaAs sample is decomposed with aqua regia (1+1). To prevent Ga(III) interference on the ETAAS determination of boron, a double extraction of the chlorogallic acid (HGaCl4) in diethyl ether is performed. To improve the overall ETAAS performance, the graphite tubes were pre-treated with iridium(III) and tungsten(IV). A mixed chemical modifier containing Ni(II), Sr(II) and citric acid was also used. The characteristic mass (m0) is 301 +/- 47 pg and the detection limit (3sB) is 2.4 microg g(-1). The classic UV-Vis spectrophotometric procedure using curcumin was also extended to the determination of boron in GaAs. By masking Ga(III) with EDTA and a preliminary extraction of boron with 2-ethyl-hexane 1,3-diol, performed on a semi-micro scale, a detection limit of 0.6 microg g(-1) was achieved. Both methods were applied to the analysis of two Si-doped GaAs samples which were suspected of being boron-contaminated. Results are compared with those obtained by direct analysis of the decomposed sample solution using the inductively coupled plasma atomic emission spectrometry (ICP-AES). PMID- 12556029 TI - Presence and racemisation of amino acids in calcium oxalate patinas. A case study from the Baptistery in Parma, Italy. AB - The organic material present in oxalate patinas, collected from the stone external surfaces of the Baptistery in Parma (Italy), was analysed by GC-MS. The high racemisation levels determined for several specific amino acids indicate that microbial activity could contribute to the formation and modification of the organic material leading to the calcium oxalate patinas. PMID- 12556030 TI - Thermodynamic parameters for the binding of sulfate by open chain polyammonium cations. AB - The interaction of sulfate with some open chain polyammonium cations was quantitatively studied by potentiometric (H(+)-glass electrode) measurements. Enthalpy changes, for some systems, were determined calorimetrically. Some literature data were also considered in the discussion of results. In all the systems, A(SO4)Hi(i-2) species are formed (i = 1, ...m; m = maximum protonation degree of amine), and, for i > or = 4, also A(SO4)2Hi(i-4) species were found. Both the stability and the formation enthalpies, for these complexes, are a direct function of the charges of reactants, and some empirical relationships are reported. A small, but significant, difference was found between the stability of unsubstituted and N-alkylsubstituted polyammonium cation-sulfate complexes. PMID- 12556031 TI - Quantitative determination of fluoride as a major pollutant in the emissions from the thermal treatment of clayey raw materials. AB - Three clayey raw materials for production of traditional ceramics, all with medium to high fluorine content, have been investigated for determining their attitude to fluoride emission during firing, at varying parameters of the firing cycle, maximum firing temperature included, and in the presence or in the absence of calcium carbonate either added to the sample or naturally present in it. Different analytical methods for the determination of fluoride have been tested, together with different sample preparation procedures aimed to obtain minimum loss of fluoride in the pre-treatment step. PMID- 12556032 TI - Heavy metals content in rain water collected in three different locations in Italy during 1999. AB - Rain water samples collected at three different locations in Italy during 1999 were analysed for heavy metals determination. Results for V, Cr, Mn, Co, Cu, Cd, Sb, Pb, Al and Ni are reported in this paper. For most of the elements higher concentration values have been measured in the samples from Alpe Gera (rural site): only for Cd and V were the concentrations higher at Pisa. The total annual deposition (microg m(-2) y(-1)) shows a similar behaviour, with Alpe Gera being the location with highest total annual input for Cr, Mn, Co, and in particular for Cu, Sb, Pb and Al. The reason of the higher deposition at a rural site in comparison to a urban one (Milan) is not known: further experimental work and investigations are needed. Generally, no evident seasonal trends have been observed for the measured concentrations, while few correlations have been found among elements within the same station. Crustal contribution to total concentration varies from about 20% for V to less than 1% for Cu, Cd, Sb, Pb and Ni. PMID- 12556033 TI - Effect of lipophylic salts on ise detection limit: application to calixarene based highly efficient potassium-selective electrodes. AB - A new interpretative model for interpolation of potentiometric data from liquid membrane ion selective electrodes has been formulated; the model allows one to achieve information about partitioning equilibria establishing at the membrane analysed solution interface of hydrophilic cations present in the membrane as counterions of exchanger lipophylic anions. The fundamental importance of such equilibria is related to their contribution to the detection limit of the electrodes. The soundness of the proposed model was positively verified by employing it in the characterization of new K(+)-selective electrodes based on a calixarenic ionophore. Although the validity of interpolation based on our model does not significantly differ from that obtained with the simpler model providing the inclusion of a constant parameter in the logarithmic argument of Nicolski Eisenman's equation, the information obtained resulted to be analytically suitable for optimising the membrane composition and, as a consequence, the performance of the electrodes. PMID- 12556034 TI - A preliminary study on the migration mechanism of heavy metals into the vadose zone of soils of a dismissed industrial site. AB - Within the framework of the site assessment of a dismantled steel-making plant area contaminated by Electric Arc Furnace dust (EAF dust), the actual migration of Cr, Cu, As, Cd, Pb, Zn and Ni (contaminants) from an industrial waste layer towards the vadose zone of the underlying soil has been evaluated. The crystalline phases and contaminant abundance of samples of the industrial waste layer and the surrounding soils were analysed, while the release of contaminants was evaluated by leaching tests. Results indicate the migration of pollutants from the industrial waste layer into the subsurface soil, but the high leachability of soil hides its real degree of contamination, with the consequent possibility of underestimating the hazardous characteristics of the actual situation. PMID- 12556035 TI - Determination of some trace metals in waters by flame atomic absorption spectrometry after preconcentration on Amberlite XAD-16 resin with sodium tetraborate. AB - A method was described for the determination of the elements Cr, Mn, Fe, Co, Ni, Cu, Cd, Pb, and Bi in waters by flame atomic absorption spectrometry (FAAS) after separation and preconcentration on Amberlite XAD-16 resin with sodium tetraborate using a chromatographic column. Parameters influencing the analytical performance, including pH and the volume of sample, amount of analyte and interfering effect of co-existing ions, were studied in detail. The recovery values were quantitative (> or = 95%), and the relative standard deviation (RSD) and detection limit (DL) varied in the range of 1.1-2.4% (n=10) and 0.002-0.177 microg m(-1) (3s, n=20), respectively. After being optimized, the proposed method was applied to the drinking water, waste water and artificial sea water samples. Recovery values of the elements investigated, were quantitative for tap water and synthetic sea water, except for Mn, Co and Ni (including also Cd for synthetic sea water). Recovery values of Cd, Pb, Cu and Co were found to be 95, 102, < or = 87, and < or = 83%, respectively, for the waste water samples. PMID- 12556036 TI - Comparison of microwave digestion procedures for the determination of some elements in asphaltite ash using ICP-AES. AB - A microwave digestion procedure for asphaltite ash was developed in an attempt to facilitate routine analysis and obtain reproducible conditions or comparable results. The conditions of the most effective procedure for 0.1 g asphaltite ash samples are 1 ml of HNO3 + 3 ml of HCl + 1 ml of HF + 1 ml of deionized water as acid mixture and 15 min for digestion time. The digestion was accomplished in five stages applying continuously 90% to 20% of the microwave power and 20 to 100 psi of the pressure for 10 min of total time. Concentrations of selected elements, Cr, Co, Fe, Mn, Ni and Zn were measured using inductively coupled plasma-atomic emission spectroscopy. The proposed method of digestion provided precise results with relative standard deviations generally less than 3% for investigated elements. Results for fly ash as standard reference material was in good agreement with certified values. PMID- 12556037 TI - Trace analysis of 1,10-phenanthroline based herbicide in water and soil samples. AB - A sensitive analytical procedure for diquat herbicide viz. 5,6 dihydropyrazino[1,2,3,4-lmn]-1,10-phenanthrolinium dichlorides in drinking water and soil samples, using differential pulse adsorptive stripping voltammetry (DPAdSV) has been developed. This involved the adsorptive accumulation of a diquat herbicide onto a hanging mercury drop electrode [accumulation potential 0.8 V vs Ag/AgCl, accumulation time 60 s, pH 7.0, ionic strength 0.1 M] for DPAdSV measurement. The detection limit for diquat herbicide was as low as 0.034 micromol L(-1) level (S/N 2, RSD 2.9%) at an accumulation time of 60 s. PMID- 12556038 TI - Selective recovery of gold from secondary sources by Dowex-50-X8 sorbed Rhodamine B resin. AB - Gold recovery from spent materials has been performed on modified cation-exchange resin. The modified resin was prepared by sorption of Rhodamine-B on Dowex-50-X8 (Na+) resin at pH 2.0. Distribution coefficients of gold ion have been determined with 15% (w/v) potassium chloride in different concentrations of hydrochloric acid. On the basis of Kd values quantitative separation of gold from secondary sources has been achieved. Gold(III) has been selectively separated from gold containing spent materials by column chromatography using modified ionexchange resin and the recovery was more than 92% compared to conventional cyanidation process. The results of the proposed method are suggestive of its applicability to other gold containing secondary sources. PMID- 12556039 TI - Simple and effective titrimetric procedure for the determination of sulfide and cysteine in the leather industry effluents. PMID- 12556040 TI - A new method for the ascorbic acid assay using iron(II)-pyridine-2,6-dicarboxylic acid complex. PMID- 12556041 TI - Determination of palladium and phosphorus in biologically active palladium(II) complexes of aminophosphonic acid derivatives. PMID- 12556042 TI - Sorption and preconcentration of some trace metals on sepiolite. PMID- 12556043 TI - Investigations on genetically modified maize (Bt-maize) in pig nutrition: fattening performance and slaughtering results. AB - A grower finisher performance trial with forty-eight pigs was designed to compare the growth performance of pigs fed diets containing either genetically modified (GM) Bt-maize (NX6262) or its parental maize (Prelude) line. During the experiment, the pigs were fed with a grower and a finisher diet both containing 70% maize investigated in a previously study which showed that they contained similar ME values and digestibility of crude nutrients. The pigs with an initial live weight of 23.9 +/- 3.0 kg were allotted to single boxes. During a 91 days growing period the pigs of both groups recorded equal performance in daily weight gain (AW) 815 +/- 93 vs. 804 +/- 64 g/d depending on equal amounts of feed intake 1.95 +/- 0.15 vs. 1.94 +/- 0.15 kg/d (parental vs. transgenic). The results confirm equal performance among growing-finishing pigs fed parental or genetically modified maize containing diets. For slaughtering the pigs were divided into 4 groups with a different duration of the finishing period. After slaughtering the carcass characteristic were registered. PMID- 12556044 TI - Effect of dietary supplementation with alpha-galactosidase preparation and stachyose on growth performance, nutrient digestibility and intestinal bacterial populations of piglets. AB - Two experiments were conducted to investigate the effect of dietary supplementation with alpha-galactosidase preparation and stachyose on growth performance, nutrient digestibility and intestinal bacterial populations of piglets. In Experiment I 72 crossbred piglets were allotted to three treatments, i.e. 1) control (basal) diet, 2) alpha-Gal1 and 3) alpha-Gal5, the basal diet supplemented with 0.01% and 0.05% alpha-galactosidase preparation (alpha-Gal), respectively. Average daily gain, average daily feed intake and feed conversion ratio were not influenced by alpha-Gal supplementation. In Experiment II 72 crossbred weanling piglets were randomly assigned to four treatments with a complete factorial design, i.e. 1) basal diet, 2) basal diet with 0.01% alpha-Gal supplementation, 3) basal diet with 1% stachyose supplementation and 4) basal diet with 1% stachyose and 0.01% alpha-Gal supplementation. Average daily gain and feed conversion ratio were influenced by alpha-Gal and stachyose supplementation. The ileal digestibility of stachyose, raffinose, gross energy and crude protein was improved significantly by alpha-Gal supplementation. The microbial populations in the intestine were modified by both alpha-Gal and stachyose supplementation. The data suggested that stachyose supplementation had an adverse effect on the growth performance of piglets and alpha-Gal supplementation could ameliorate it, especially in younger pigs. PMID- 12556045 TI - Determination of apparent ileal amino acid digestibility in rapeseed meal and cake processed at different temperatures using the direct and difference method with growing pigs. AB - Studies were conducted with ten barrows, average initial body weight 34.5 +/- 2.1 kg, fitted with a T-cannula at the distal ileum, to study the accuracy of determination of the apparent ileal digestibility (AID) values of crude protein (CP) and amino acids (AA) in rapeseed meal and cake and the effects of processing, using the difference method. Five corn starch-based diets in the studies were formulated to contain 17.7% CP and based on soybean meal, prepress extraction rapeseed meal, prepress-extraction rapeseed meal plus soybean meal, high-temperature press rapeseed cake plus soybean meal, or low-temperature press rapeseed cake plus soybean meal as the sole source of dietary protein. The design was an incomplete Latin Square involving two three-week periods and five treatments. It was found that the AID values of CP and most AA determined with the difference or direct method were significantly lower in rapeseed meal or cakes than soybean meal. The AID values of CP and most AA in prepress-extraction rapeseed meal, high-temperature press or low-temperature cakes determined with the difference method were no difference from those in prepress-extraction rapeseed meal determined with the direct method. The AID values of CP and AA in rapeseed meal and cake determined with the difference method were accurate, when the contribution of CP and AA from rapeseed was more than 50%. The AID values of CP and AA (especially lysine) were lower in the high-temperature press rapeseed cake than in the low-temperature press cake or the prepress-extraction meal. PMID- 12556046 TI - Feed intake of young double-muscled bulls fed on grass and supplemented with sugar-beet pulp. AB - Daily dry matter intake in young growing double-muscled bulls, fed indoors on grass, was estimated based on forty-four intake data from 28 animals, ageing at least five months and weighing up to 400 kg live weight. Intake was measured during five consecutive days using one of eighteen cuts of grass. Fresh meadow grass (mainly Lolium perenne) was fed ad libitum and two kg dried sugar-beet pulp was offered per animal and per day. Animal live weight averaged 278 +/- 82 kg and mean total daily dry matter intake amounted to 5.05 +/- 1.59 kg or 73.6 +/- 13.7 g per kg metabolic weight, while pulp dry matter intake amounted to 1.49 +/- 0.50 kg per day. Regression analysis showed that animal as well as feed characteristics could explain up to approximately 90% of the variation in daily dry matter intake. The supplementation resulted in an extra daily dry matter intake of 0.68 g per g pulp dry matter. Intake of double-muscled animals was considerably lower than previously reported for non-double-muscled cattle. An extra supplementation of young grazing double-muscled animals could be advised from these findings, while extra protein should also be considered. PMID- 12556047 TI - Performance of growing-finishing pigs fed diets based on normal or low viscosity rye fed with and without enzyme supplementation. AB - One hundred and forty crossbred pigs (Pig Improvement Canada Ltd, Acme Alberta), weighing an average of 21.5 + 3.6 kg, were assigned on the basis of sex, weight and litter to one of five dietary treatments. The control diet was barley-based, while in the remaining four treatments, the grain component consisted of either normal or low viscosity rye fed with or without pentosanase. Digestibility coefficients for dry matter, crude protein and gross energy were significantly (P = 0.0001) higher for the rye-based diets than the barley based diet. There were no differences in digestibility coefficients between normal and low viscosity diets or between diets supplemented or unsupplemented with pentosanase. Over the entire experimental period (21.5 to 100.7 kg), pigs fed the normal viscosity rye gained better (P = 0.001) and had a higher daily intake (P = 0.001) than pigs fed the low viscosity rye. Pigs fed the low viscosity rye tended to gain slower than pigs fed barley and with lower intake while pigs fed the normal viscosity rye had similar gains and intake to the pigs fed barley. Enzyme supplementation failed to improve pig performance. Males had higher gains and intake than females (P < 0.05). Enzyme supplementation, rye viscosity or choice of cereal grain had no effect on any of the carcass traits measured. In conclusion, the results of the present experiment indicate that rye is a good alternative to barley for use in growing-finishing rations and becomes increasing attractive as the age of the pig increases. Breeding efforts directed towards reducing the viscosity of rye are unlikely to be successful in improving the nutritive value of rye for swine. PMID- 12556048 TI - Research note: Postnatal development of electrolyte transport in calf rumen as affected by weaning time. AB - In a previous study we found a positive correlation between early weaning in calves and morphological parameters which were indicative of ruminal development, i.e. the length and width of the papillae. The objective of the present study was to determine to what extent this observation could be reflected by modulations of absorptive and secretory functions of the rumen mucosa. For this purpose the short-circuit currents (Isc) as a measure of electrogenic net ion fluxes and the transepithelial conductances (G(T)) as a measure of the overall tissue permeability were measured in vitro applying the Ussing-chamber technique. Simultaneously, the unidirectional flux rates of sodium and chloride across rumen wall epithelia were determined in the absence of electrochemical gradients. Under these conditions, significant positive net flux rates (Jnet) clearly indicate active mechanisms for electrolyte absorption. For the experiments 12 male Holstein calves 7 d of age were assigned to three groups of 4 animals each: milk group (I, slaughtered after 6 weeks of age), late weaning group (II, slaughtered after 9 weeks of age) and early weaning group (II, weaned after 6 weeks of age and slaughtered after 9 weeks of age). Whereas G(T) values remained unaffected by different age and feeding, Isc values were significantly affected by early weaning but were not influenced by age. Irrespective of weaning time active absorption of Na+ tended to be higher by about 60% in 9 weeks old animals. Active absorption of chloride was significantly increased in milk fed 9 weeke old calves and this effect was further stimulated by early weaning. In conclusion, the data show an increasing active Na+ absorption with age in calf rumen that could not be influenced by early weaning. Similarly, active Cl- absorption was initially increased during postnatal development and this effect could be stimulated further by early weaning. PMID- 12556049 TI - Sol Silverman receives the 2002 Margaret Hay Edwards Achievement Award. PMID- 12556050 TI - Predictors of self-perceived competency in cancer screening examinations. AB - BACKGROUND: Factors contributing to medical students' self-perceived competency in cancer screening examinations have not been well studied. METHODS: Ending third-year students of UCLA Medical School were surveyed to assess their self perceived competency and training in performing clinical breast, Pap smear, digital rectal, and skin cancer examinations. Analysis of variance was conducted to compare the differences in competency ratings. Multiple regression analysis was applied to identify predictors of competency. RESULTS: Fifty-two (67%) of the 78 third-year medical students sampled participated in the study. They reported being least (p < 0.001) competent in performing skin cancer examinations. The number of times practicing screening examinations was the most significant (p < 0.001) educational predictor for self-reported competency in every area except skin cancer. More practice is needed in each screening exam before students feel somewhat or extremely competent. CONCLUSION: Student hands-on experience is crucial in the development of cancer screening competency. PMID- 12556051 TI - Use of a computerized risk-appraisal instrument for cancer prevention education of medical students. AB - BACKGROUND: Interactive computer-based tools are increasingly used for patient and medical student instruction. METHOD: To assess medical students' responses to a computer-based risk-appraisal instrument aimed at teaching concepts of risk and cancer prevention, and to evaluate perceived barriers to using such tools in clinical practice and knowledge about risk factors for colon cancer. DESIGN: An electronic survey of 133 second-year medical students elicited their responses to the instrument after using it for a case-based learning exercise. RESULTS: The students identified several potential barriers to use, including inadequate time during the office visit (75% of students) and problems with literacy (48%). They were generally knowledgeable about risk factors for colon cancer. 84% found the instrument "very helpful" or "somewhat helpful" in understanding a case patient's health and setting priorities for behavioral changes. CONCLUSIONS: Medical students may benefit from exposure to interactive, computer-based tools such as health-appraisal instruments when learning skills in risk counseling and cancer prevention. Second-year students were already concerned about having adequate time during office visits to use such instruments. PMID- 12556052 TI - Assessment of a lecture on cancer prevention and the early detection of cancer. AB - BACKGROUND: Cancer prevention and the early detection can affect morbidity and mortality. Through educational programs, recommendations for beneficial lifestyle changes and cancer screening may be introduced to the public. The purpose of this study was to determine whether a videotaped lecture concerning cancer prevention and early detection is of educational value. METHOD: College students in a health science class participated in the study. The students' comprehension of the subject matter was assessed immediately before and a week after they viewed the lecture. RESULT: The students' scores on the second test were significantly better as measured by a paired-difference experiment. CONCLUSION: This videotaped lecture has merit as an educational program. PMID- 12556053 TI - Medical school exposure to spirituality and response to a hypothetical cancer patient. AB - BACKGROUND: Spiritual and religious issues (SRI) in medical school curricula may promote psychosocial and spiritual sensitivity, but few data exist on this relationship. METHOD: A questionnaire was administered to third-year medical students (response rate = 69.2%). Students indicated exposure to SRI and read a vignette about a hypothetical cancer patient. RESULTS: SRI exposure through lectures, small-group discussions, and physician modeling predicted, respectively, likelihood of extra attention toward the patient, conversing with the patient about dying, and praying with the patient. CONCLUSIONS: Exposure to SRI in medical school may sensitize students to the psychosocial and spiritual needs of dying patients. PMID- 12556054 TI - Perceptions of tongue lesions by dental hygiene students and otolaryngologists. AB - BACKGROUND: Consistent perceptions of oral lesions by all professionals who examine the mouth would improve the poor survival rates from oral cancer through earlier diagnosis. METHODS: Two techniques from perceptual psychology measured how 14 otolaryngologists and 85 dental hygiene students judged differences among seven tongue lesions. Multidimensional scaling produced a perceptual "map" showing the underlying dimensions in these judgments. Paired comparisons showed rankings of concern. RESULTS: Otolaryngologists rated the lesions on the basis of size and depth; dental hygiene students on the basis of color and size. Otolaryngologists were more consistent than students, both within and between observers. The ranking of lesions from best to worst was different in the two groups. CONCLUSION: Two groups of health-care professionals attended to different dimensions in making decisions about oral lesions. Variations of these tests could evaluate or instruct students in diagnostic skills relevant to oral cancer. PMID- 12556055 TI - Development and implementation of a curriculum in communication skills and psycho oncology for medical oncology fellows. AB - BACKGROUND: Over a ten-year period, the authors developed and implemented a disease-trajectory-based curriculum in communication skills and psycho-oncology for oncology fellows. METHODS: The basic format is a weekly ward round centered on a patient interview and subsequent discussion. More recently, this has been supplemented by a written curriculum. RESULTS: Former fellows and current trainees perceive this training o be an important component of their overall training. CONCLUSION: This kind of curriculum is one effective model for teaching communication skills and psycho-oncology to medical oncology trainees. PMID- 12556056 TI - Graduate education in oncology nursing for minorities. AB - BACKGROUND: Cancer statistics reveal disturbing morbidity and mortality rates among minorities, especially African Americans. A program to recruit and train minority nurses as Oncology Advanced Practice Nurses was developed at the University of Pennsylvania School of Nursing. METHODS: Since 1992, 30 African American, five Asian/Pacific Islander, and five Hispanic nurses have been supported during advanced oncology nursing study. RESULTS: Graduates have assumed positions of clinical and academic leadership in oncology nursing. CONCLUSIONS: This project strengthened the ability of a graduate program in oncology nursing to respond to needs related to the education of minority students and to the care of minority populations with cancer. PMID- 12556057 TI - Variations in approaches to breast cancer screening among primary care physicians. AB - BACKGROUND: Guidelines for breast cancer screening often provide conflicting recommndations. METHODS: A cross-sectional survey design was used to examine approaches to cancer screening among primary care physicians. RESULTS: Among the 187 respondents, levels of agreement with the "correct" response ranged between 50% and 73% for each of the four items relating to breast cancer screening; only 42% of physicians demonstrated agreement with three or more of the four items. Physician gender, specialty group, and age category were significant predictors of responses. CONCLUSIONS: These findings suggest the need to implement educational interventions as one means of decreasing variation in breast cancer screening. PMID- 12556058 TI - A qualitative exploration of health perceptions and behaviors among adult survivors of childhood cancers. AB - BACKGROUND: Adult survivors of childhood cancer are a growing population. The objective of this study was to draw upon qualitative methods to learn more about survivors' perceived health risks and health behaviors. METHODS: Three focus groups and nine individual interviews were conducted. RESULTS: Participants' responses varied in meaning of survivor status, relatedness to other survivors, and health risk comparisons to non-survivors. Survivors expressed uncertainty about control over their health. CONCLUSIONS: The identity, health perceptions, and behaviors of adult survivors of childhood cancer are complex. Health promotion programs for survivors should be tailored to individual health risk status and should include a physician-delivered component. PMID- 12556059 TI - Appraisal of communication skills and patients' satisfaction in cross-language encounters in oncology practice. AB - OBJECTIVE: Communication skills are the most important determinant of patients' satisfaction with care. Data about the adverse effects of cross-language encounters are scarce. This prospective study was designed to examine the association between the communication language and patients' satisfaction in oncology practice. PATIENTS AND METHODS: The Art of Medicine questionnaire was used to assess patients' perceptions of clinicians' communication behaviors and patients' global satisfaction. Mean scores of patients cared for by Arabic speaking oncologists were compared with those of patients who had communications translated from English by interpreters. RESULTS: 255 patients were eligible. Patients' median age was 44 years (95% CI, 42.8-46.2). Communication was in Arabic and interpreted English for 136 (53%) and 119 (47%) patients, respectively. The two groups were comparable for various demographic and clinical variables. On a nine-point scale, mean scores for the eight questionnaire items ranged from 6.24 (95% CI, 5.91-6.56) to 8.24 (95% CI, 8.03-8.45). There was no significant difference in communication skills between Arabic- and English speaking clinicians for any questionnaire item. Moreover, a multiple regression analysis failed to identify any variable that independently influenced overall patients' satisfaction with the delivered care. CONCLUSIONS: The findings do not support a disadvantageous effect on interpersonal skills and patients' satisfaction as a result of cross-language communication. PMID- 12556060 TI - Hepatitis B knowledge and practices among Chinese American women in Seattle, Washington. AB - BACKGROUND: Chinese Americans have higher rates of chronic hepatitis B virus (HBV) infection, resulting in higher incidences of liver cancer, compared with the general population. METHODS: The authors conducted a community-based survey of HBV knowledge and preventive practices among Chinese American women in Seattle, Washington, during 1999. RESULTS: Less than half (46%) of the respondents knew that HBV could cause liver cancer. Only 35% reported that they had been serologically tested for HBV, and most of those who knew they were susceptible (61%) had not been vaccinated. CONCLUSIONS: The findings indicate low levels of HBV knowledge, screening, and vaccination among Chinese immigrants. Targeted interventions are needed to reduce the burden of HBV infection and liver cancer in this community. PMID- 12556061 TI - Storytelling as a method to teach African American women breast health information. AB - BACKGROUND: The purpose of this study was to explore the use of storytelling as a method to teach breast health information to older African American women. METHODS: A qualitative design utilizing the focus-group method. RESULTS: Six story types (categories) emerged from the focus-group data and were integrated into the breast health education class. Categories were: 1) loss, 2) pain, 3) suffering, 4) fear and worry/stress, 5) death, and 6) faith in God and prayer. Three major outcomes resulted from the storytelling format: 1) peer teaching enhanced learning, 2) incorrect information was identified and clarified, and 3) personal experiences were validated. CONCLUSIONS: Integration of African American women's self-identified issues and concerns with breast health messages through storytelling resulted in a meaningful teaching-learning experience. PMID- 12556062 TI - A breast cancer education and on-site screening intervention for unscreened African American women. AB - BACKGROUND: This study tested a breast cancer education intervention for African American women with poor screening histories. Disparities in rates of later-stage disease and death may be related to lower screening rates due to behavioral (e.g., knowledge gap) and structural (e.g., access) barriers. METHODS: 94 African American women (low-income, poor screening histories) were randomly assigned for an educational and on-site screening intervention, or non-intervention status. Primary care sites were used for educational purposes. Three-month post intervention mammography was assessed. RESULTS: The intervention group had significantly higher screening rates than the non-intervention group. The latter had a knowledge gap, poor screening history, cancer fears, and insurance differences relative to the screened women. Screening was predicted by behavioral, rather than structural factors. CONCLUSIONS: Since low-income African American women face behavioral and structural barriers to breast cancer screening, an educational program that improves knowledge levels and is offered in primary care sites can effectively impact screening. PMID- 12556063 TI - The Book of Job: a 2,500-year-old current guide to the practice of oncology: the nexus of medicine and spirituality. AB - GOALS: To establish the role of ancient literature and religious tradition to the modern practice of oncology; foster awareness of practicing in a historical context resulting from different traditions; and propose a spiritual context for the practice of oncology and explore methods to highlight this perspective in cancer education. METHOD: Contextual and content analysis of a religious text shared by the most common religious traditions of the West (Christianity, Judaism, and Islam). RESULTS: The origin of suffering eludes all logical explanations. All religious traditions affirm that the sufferer should be heard, cared for, and kept part of the human consortium, and under no circumstances blamed for the disease. In terms of oncology practice this means that the treatment should be negotiated with the patient according to his or her need; that physicians' obligations for care continues after the treatment fails, and that patients' lifestyles or poor compliance should not be blamed for poor outcomes. CONCLUSIONS: The Book of Job supports a spiritual perspective in oncology practice, indicating that patient care is a holistic endeavor. This perspective is the key to dealing with common interactive problems, such as adversarial relations between patient and provider in face of death and suffering, and more important, may promote care beyond treatment of the disease. PMID- 12556064 TI - What is an oncologist: a "patient phenomenology.". PMID- 12556065 TI - Increasing mupirocin resistance of Staphylococcus aureus in CAPD--should it continue to be used as prophylaxis? AB - This editorial summarizes the mechanisms of mupirocin resistance and the conditions that promote its development. While there is increasing evidence that high-level mupirocin resistance is developing during systematic use of mupirocin as prophylaxis against Staphylococcus aureus, the appearance of resistance does not preclude the beneficial effect of regular prophylaxis. Vigilance has to be exercised to notice larger-scale resistance that may require limiting the use of mupirocin. PMID- 12556066 TI - Increased leukocyte rolling in newly formed mesenteric vessels in the rat during peritoneal dialysis. AB - OBJECTIVE: Long-term peritoneal dialysis (PD) is associated with the development of functional and structural alterations of the peritoneal membrane. The present study reports the effects of chronic exposure to PD fluid on mesenteric leukocyte endothelium interactions, using intravital video microscopy. METHODS: Rats (n = 7) received 10 mL lactate-buffered 3.86% glucose-containing PD fluid daily during a 5-week period via a subcutaneously implanted mini access port that was connected via a catheter to the peritoneal cavity. In a first control group (n = 8), catheters were implanted but no fluid was instilled; a second control group (n = 8) remained untreated. The number of rolling and adherent leukocytes as well as blood flow and other fluid dynamic variables were analyzed in mesenteric postcapillary (diameter 10-25 mu) and collecting (diameter 26-40 mu) venules. Neovascularization was semiquantitatively assessed after inspection of video images and by light and electron microscopy. Using FITC-labeled albumin, microvascular leakage was examined. RESULTS: Rats exposed to PD fluid showed a more than twofold increase in the number of rolling leukocytes (p < 0.01); the number of adherent leukocytes was not changed. Furthermore, exposure to PD fluid induced severe neovascularization in rat mesentery. No microvascular leakage was observed in the various groups. The observed differences could not be explained by differences in systemic or local hemodynamic parameters or peripheral leukocyte counts, but is most likely associated with new vessel formation. CONCLUSIONS: Exposure of rat peritoneal membrane to conventional PD fluid for 5 weeks affected local leukocyte-endothelium interactions. In addition, severe angiogenesis was induced, whereas microvascular permeability remained unaltered. PMID- 12556067 TI - Generation of TNFalpha and interleukin-6 by peritoneal macrophages after overnight dwells with bicarbonate- or lactate-buffered dialysis fluid. AB - OBJECTIVE: In order to evaluate the biocompatibility profile of a newly designed peritoneal dialysis fluid (PDF), we evaluated peritoneal leukocyte (PMphi) cytokine release following overnight in vivo dwells using standard, lactate buffered, single-chamber bag PDF (Lac-PDF) and purely bicarbonate-buffered, double-chamber bag PDF containing 34 (Bic-PDF) or 39 (Bic Hi-PDF) mmol/L bicarbonate. DESIGN: A randomized, open, crossover clinical trial with single weekly test dwells was performed in stable, long-term continuous ambulatory PD patients (n = 8). During 8-hour overnight dwells, PMphi were exposed to different PDF containing 1.5% glucose. After drainage, peritoneal cells were isolated and incubated with RPMI 1640 medium for 2 or 3 hours, with and without stimulation by lipopolysaccharide (LPS). Ex vivo release of tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 was measured by specific ELISA technique. RESULTS: After pre-exposure to Lac-PDF, PMphi generated 242 +/- 279 pg TNFalpha/10(6) cells and 157 +/- 105 pg IL-6/10(6) cells. When pre-exposed to Bic-PDF and Bic Hi-PDF, TNFa and IL-6 production of PMphi was not significantly different from Lac-PDF. After LPS stimulation (100 ng/mL), PMD secretion of TNFalpha and IL-6 pre-exposed to three PDF revealed no significant differences between groups: TNFalpha was 2,864 +/- 1,216, 2,910 +/- 1,202, and 3,291 +/- 558 pg/10(6) cells after overnight dwells with Lac-PDF, Bic-PDF, and Bic Hi-PDF, respectively. Comparably, LPS stimulated (100 pg/ mL) PMphi showed IL-6 secretion of 891 +/- 335, 1,380 +/- 1,149, and 1,442 +/- 966 pg/10(6) cells for Lac-PDF, Bic-PDF, and Bic Hi-PDF. CONCLUSION: After long-term overnight dwells, initial pH, the different buffers, and varying glucose degradation product levels of PDF do not strongly affect PMphi function with respect to cytokine release. The lack of significant differences between fluids may result from the complete dialysate equilibration achieved during the overnight intraperitoneal dwell. PMID- 12556069 TI - Factors associated with sleep disorders in Chinese patients on continuous ambulatory peritoneal dialysis. AB - OBJECTIVE: This study aimed at identifying factors that are associated with sleep disorders in Chinese patients on continuous ambulatory peritoneal dialysis (CAPD). STUDY DESIGN: Questionnaire survey. SETTING: Renal dialysis unit of a tertiary-care hospital. PATIENTS: 179 Chinese patients on CAPD. MAIN OUTCOME MEASURES: The quantity and quality of sleep were compared between CAPD patients with and without self-reported sleep disorders. Demographic variables, personality traits, behavioral factors, somatic symptoms, and clinical parameters were correlated with the presence of sleep disorders. RESULTS: The mean age of the patients was 61 +/- 12 years and the male-to-female ratio was 1:1.3. The mean duration of CAPD was 47 +/- 42 months. 110 patients (61%) had self-reported sleep disorders. Among patients with self-reported sleep disorders, difficulty falling asleep was the most frequent complaint (74.5%), followed by frequent awakening (68%) and early morning waking (68%). The mean duration of sleep among these patients was 4.6 +/- 2.2 hours, versus 7.3 +/- 1.4 hours in patients without self reported sleep disorders. Three personality traits (anxiety, worry, and sadness) and two somatic symptoms (bone pain and arthralgia) were the factors significantly associated with sleep disorders. Patients with sleep disorders also more frequently reported certain symptoms suggestive of sleep apnea. Demographic variables, behavioral factors (e.g., caffeine or alcohol intake), and clinical parameters (e.g., adequacy of dialysis, body mass index, hemoglobin, albumin, parathyroid hormone levels) did not differ between patients with and without sleep disorders. CONCLUSIONS: Sleep disorders are prevalent among Chinese CAPD patients. Personality traits (anxiety, worry, and sadness) and somatic symptoms (bone pain and arthralgia) were the factors significantly associated with sleep disorders in this patient population. Behavioral factors, demographic variables, and clinical parameters did not appear to predict sleep disorders. Further studies evaluating the impact of various psychological factors on sleep disorders among CAPD patients are warranted. PMID- 12556068 TI - Selected growth factors in peritoneal dialysis: their relationship to markers of inflammation, dialysis adequacy, residual renal function, and peritoneal membrane transport. AB - OBJECTIVES: Markers of chronic inflammation, acute-phase reactants, and growth factors may be concomitantly involved in a number of pathologic processes in the general population and uremic patients. In addition, growth factors may influence peritoneal membrane transport characteristics. However, the association between plasma growth factors, markers of chronic inflammation, and peritoneal membrane transport remains largely unknown. The aim of this study was to evaluate the relationship between plasma levels of selected growth factors [basic fibroblast growth factor (bFGF), transforming growth factor beta1 (TGFbeta1), vascular endothelial growth factor (VEGF)] and markers of chronic inflammation [interleukin (IL)-6, C-reactive protein (CRP), and fibrinogen] in continuous ambulatory peritoneal dialysis (CAPD) patients. The potential link between the above substances and dialysis adequacy was also explored. DESIGN: Single-center, cross-sectional study. SETTING: Peritoneal Dialysis Unit, Medical Faculty, Jagiellonian University Hospital, Krakow, Poland. PATIENTS: 32 stable end-stage renal disease patients (13 M, 19 F; mean age 53.6 +/- 13.7 years) on CAPD for a median period of 19.5 months. Patients free from signs and symptoms of any inflammatory disease (including peritonitis) for at least 3 months were included into the study. All patients underwent measurements of dialysis dose [Kt/V, weekly creatinine clearance (wCCr)] and peritoneal solute transport using a standard peritoneal equilibration test (PET). METHODS: TGFbeta1, bFGF, VEGF, and IL-6 were measured with ELISA, CRP was assayed with immunonephelometry, and fibrinogen with Multifibren U reagent (Dade Behring Marburg GmbH, Marburg, Germany). Nephron 97 for Windows software was used to assess dialysis adequacy. RESULTS: Significant positive correlations between plasma bFGF and IL-6, as well as fibrinogen concentrations (R = 0.36, p < 0.05 and R = 0.39, p < 0.05, respectively), were found. VEGF correlated significantly with IL-6 and CRP (R = 0.65, p < 0.0001 and R = 0.51, p < 0.005, respectively). An association between VEGF and bFGF was also found (R = 0.59, p < 0.0005). Serum level of TGFbeta1 revealed no relationship with any marker of acute-phase activation, remaining growth factors, or dialysis adequacy. Positive correlation between TGFbeta1 concentration and dialysate-to-plasma ratio for creatinine in PET (R = 0.35, p < 0.05) was found. In addition, patients with lower solute transport (low/low average transporters) had lower serum levels of both bFGF and TGFbeta1 compared to patients with higher solute transport. Patients with total wCCr > 60 L/ week/m2 were characterized by lower levels of bFGF and IL-6. Serum level of IL-6 and plasma levels of bFGF and VEGF were significantly lower among subjects with residual renal function (RRF) > 2.0 mL/minute. CONCLUSIONS: Our results indicate that systemic inflammation in peritoneal dialysis patients is associated with increased plasma VEGF and bFGF but not TGFbeta1. The negative correlation with RRF suggests that either the renal clearance of these cytokines and growth factors may contribute to their elimination, or cytokines and growth factors have a negative impact on RRF. We also suggest an association between serum levels of growth factors tested and peritoneal membrane function. PMID- 12556070 TI - Assessment of fluid status in peritoneal dialysis patients. AB - OBJECTIVES: To assess the influence of abnormalities in fluid status and body composition on agreement between multifrequency bioimpedance analysis (MF-BIA), segmental BIA (sigmaBIA), the Watson formula, and tracer dilution techniques. DESIGN: Cross-sectional. SETTING: Multicenter. PATIENTS: 40 patients (29 males, 11 females) on peritoneal dialysis (PD). MAIN OUTCOME MEASURES: Agreement between the various techniques used to assess total body water (TBW) [MF-BIA, deuterium oxide (D2O), and the Watson formula] and extracellular water (ECW) [MF-BIA, bromide dilution (NaBr), and sigmaBIA], also in relation to the relative magnitude of the body water compartments [ECW (NaBr):body weight (BW) and TBW (D2O):BW] and body composition (DEXA). Second, the relation between body water compartments with echocardiographic parameters. RESULTS: Wide limits of agreement were observed between tracer dilution techniques and MF-BIA [TBW (D2O - MF-BIA) 2.0 +/- 3.9 L; ECW (NaBr - MF-BIA) -2.8 +/- 3.9 L], which were related to the relative magnitude of the body water compartments: r = 0.70 for ECW and r = 0.40 for TBW. sigmaBIA did not improve the agreement [ECW (NaBr-sigmaBIA): 3.7 +/- 2.9 L]. Also, wide limits of agreement were observed between D2O and the Watson formula (-2.3 +/- 3.3 L). The difference between D2O and Watson was related to hydration state and to percentage of fat mass (r = 0.70 and r = -0.53, p < 0.05). Both ECW and TBW as assessed by BIA and tracer dilution were related to echocardiographic parameters. CONCLUSION: Wide limits of agreement were found between MF-BIA and sigmaBIA with dilution methods in PD patients, which were related to hydration state itself. The disagreement between the Watson formula and dilution methods was related to both hydration state and body composition. PMID- 12556071 TI - Increasing the dialysis volume and frequency in a fixed period of time in CPD patients: the effect on Kpt/V and creatinine clearance. AB - BACKGROUND: The National Kidney Foundation Kidney Disease Outcomes Quality Initiative (K/DOQI) has evidence- and opinion-based recommendations for weekly Kt/V(urea) and weekly total creatinine clearance (CC) in chronic peritoneal dialysis (CPD) patients. Using standard continuous ambulatory peritoneal dialysis technique, it is often difficult to achieve the suggested targets in anuric patients with large body mass. Thus, the use of automated peritoneal dialysis (APD) has been increasingly utilized to achieve adequate clearances. Automated dialysis is usually performed at night over an 8- to 10-hour period. The role of increases in dialysate volume and frequency of exchanges during this time period to achieve these target K/DOQI recommendations remains uncertain. We decided to study the effects of increasing the volume and number of exchanges in a fixed period of time in CPD patients. METHODS: In the New Haven CAPD unit, 29 patients maintained on APD were considered eligible for the study and 11 agreed to participate. The patients were characterized according to standard peritoneal equilibration test criteria. The patients were placed into two groups: group 1 included high (H) and high-average (HA), and group 2 low-average (LA) transporters. The patients were dialyzed at night for 9 hours with standard cycling technique, using 2.5% Dianeal (Baxter Healthcare, Deerfield, Illinois, USA) solution, with a cycle volume of 2,500 mL, and a 2,000-mL daytime dwell. Three studies were done on each patient using a total dialysis volume of 9.5 L (3 cycles), 14.5 L (5 cycles), and 19.5 L (7 cycles). Daily Kpt/V(urea) and daily CCp (peritoneal) (L/day/1.73 m2) were obtained. RESULTS: Six patients were H or HA (group 1) and 5 were LA transporters (group 2). For the group 1 patients, mean weight was 86.6 +/- 13.5 kg; Kpt/V(urea) was 1.68 +/- 0.21 using 9.5 L, 2.03 +/- 0.28 for 14.5 L (p < 0.05 compared to 10 L), and 2.28 +/- 0.28 with 19.5 L (p < 0.05 compared to 10 L and 15 L); mean weekly CCp was 45.43 +/- 7.63 L/1.73 m2 for 9.5 L (p < 0.05 compared to 14.5 L and 19.5 L), 51.17 +/- 7.07 with 14.5 L, and 54.67 +/- 10.08 for 19.5 L; ultrafiltration rates were not different in the three studies. For the group 2 patients, mean weight was 74.3 +/- 17.7 kg; mean weekly Kpt/V(urea) was 1.68 +/- 0.35 using 9.5 L, 2.10 +/- 0.42 for 14.5 L (p < 0.05 compared to 9.5 L), and 2.31 +/- 0.56 for 19.5 L (p < 0.05 compared to 9.5 L and 14.5 L); mean weekly CCp was 42.56 +/- 10.64 L/1.73 m2 for 9.5 L (p < 0.05 compared to 14.5 L and 19.5 L), 50.89 +/- 12.66 for 14.5 L, and 51.94 +/- 11.20 for 19.5 L; ultrafiltration was lower in the 9.5-L study than in the 14.5-L and 19.5-L studies, but was not different in the 14.5-L and 19.5-L studies. CONCLUSIONS: In both H/HA and LA transporters, Kpt/V(urea) and CCp rise significantly when the frequency of exchanges and total volume of dialysate are increased. Thus, the use of larger volumes of dialysate with cycling peritoneal dialysis may result in increased clearances of urea and creatinine. PMID- 12556072 TI - Spatial distribution of dialysate in patients and its implications to intradialysate diffusion. AB - OBJECTIVE: To visualize and quantify the spatial distribution of dialysate in patients on continuous ambulatory peritoneal dialysis (CAPD) and, hence, estimate diffusion times for fluid "pockets" wherever intradialysate concentration gradients may not be dissipated by convective currents. DESIGN: Contrast medium was added to the dialysate of three supine CAPD patients before an exchange prior to computed tomographic (CT) scanning. Spatial information in the CT scanner was then downloaded to other computers and processed to produce impressive three dimensional models of dialysate distribution using "wire frame technology." RESULTS: Models differed between patients but all demonstrated pooling of dialysate in the paracolic gutters, subphrenic space, and, especially, in the pelvic cavity. Some pockets of fluid were almost isolated. Quantitatively, the models can account for over 80% of the volume of the exchange (2.5 L), displaying an effective area of contact of 913-450 cm2 between parietal peritoneum and dialysate. This amounts to only 11% -21% of the anatomic area, again emphasizing the uneven distribution of dialysate. Ignoring very thin (< 0.1 mm) films of dialysate, the bulk (80%) had mean thicknesses ranging from 1.6 to 1.9 cm. Transcendental equations for bulk diffusion were then applied to these findings to determine a theoretical time for urea of about 2-3 hours to half-saturation, or 5-7 hours to 80% saturation, in the absence of convective currents. CONCLUSIONS: The distribution of dialysate within the peritoneal cavity is very uneven, resulting in long diffusion times in fluid pockets wherever convective currents may be minimal. Hence, intradialysate diffusion should not be ignored when modeling peritoneal dialysis. PMID- 12556073 TI - Sodium removal in patients undergoing CAPD and automated peritoneal dialysis. AB - OBJECTIVES: To compare sodium removal in continuous ambulatory peritoneal dialysis (CAPD) and automated peritoneal dialysis (APD) patients, and to identify the main factors that modify Na removal in clinical practice in these patients. DESIGN: Study in three steps. Cross-sectional observational (Study A), and longitudinal interventional (Studies B and C). PATIENTS AND METHODS: First (Study A) we carried out a cross-sectional survey of Na removal in 63 patients on CAPD and 78 patients on APD. Second (Study B), we studied Na removal in 32 patients before and after changing from CAPD to APD therapy. Finally (Study C), we analyzed the impact on Na removal of introducing icodextrin for the long dwell in 16 patients undergoing CAPD or APD. RESULTS: In Study A, total Na removal averaged 210 mmol/day for CAPD patients and 91 mmol/day for APD patients (p < 0.001); Na removal was < 100 mmol/day in 7.1% of CAPD patients and 56.4% of APD patients. Multivariate analysis identified ultrafiltration [B = 125 mmol/day, 95% confidence interval (CI) 110,140], CAPD therapy (B = 60 mmol/day, 95%CI 37, 83), and residual diuresis (B = 51 mmol/L, 95%CI 34, 69) as independent predictors of Na removal (adjusted r2 = 0.76). For APD patients, longer nocturnal dwell times and performing a supplementary diurnal exchange were also independently associated with higher Na removal rates. In Study B, Na removal decreased from 192 to 92 mmol/day (median) after the change to APD (p = 0.02). In Study C, peritoneal Na removal increased from 98 to 148 mmol/day (median) (p = 0.04) after introducing icodextrin. CONCLUSIONS: Standard APD schedules are frequently associated with poor Na removal rates. For any degree of ultrafiltration, Na removal is better in CAPD than in APD. Icodextrin, supplementary diurnal exchanges, and longer nocturnal dwell times improve Na removal in APD. Sodium removal can be estimated from ultrafiltration in patients on CAPD, but must be specifically monitored in patients on APD. PMID- 12556074 TI - Low calcium dialysate increases cardiac relaxation in CAPD patients. AB - OBJECTIVE: To establish whether changes in serum calcium affect left ventricular (LV) function in continuous ambulatory peritoneal dialysis (CAPD) patients. METHODS: This study was conducted on 28 clinically stable CAPD patients (11 females, 17 males). Left ventricular relaxation and systolic function were echocardiographically examined in all patients during standard dialysate (containing 1.75 mmol/L calcium) treatment. All patients were then changed to low calcium dialysate (1.25 mmol/L calcium) for 1 month and all patients were re examined echocardiographically. Decrement in isovolumic relaxation time (IVRT) and deceleration time (DT), and increment in the ratio of peak early to peak late diastolic velocities (E/Amax) were admitted as indexes showing improvement in LV relaxation. 17 age- and sex-matched controls were also echocardiographically examined. RESULTS: Deceleration time, interventricular septal thickness at systole (IVSTS) and diastole (IVSTD), and posterior wall thickness at systole (PWS) and diastole (PWD) were higher in CAPD patients using standard dialysate than in normal controls. With the use of low calcium dialysate, DTs were similar but IVSTS, IVSTD, PWS, and PWD values remained higher. In normal controls, E/Amax values were higher than those in CAPD patients using standard dialysate (p < 0.001) and low calcium dialysate (p = 0.009). Serum intact parathyroid hormone level, weight, clinical volume status, and blood pressure were similar throughout the study period. Serum ionized calcium levels were decreased significantly during low calcium dialysate treatment. The changes in IVRT, DT, and E/Amax suggest improvement in LV relaxation during low calcium dialysate treatment. CONCLUSION: Left ventricular relaxation is increased with the use of low calcium dialysate compared with standard dialysate treatment. The idea of possible beneficial effects of increment in LV relaxation on cardiovascular morbidity and mortality deserves further studies. PMID- 12556076 TI - Lack of effects of low-dose aspirin on high-sensitivity C-reactive protein, hemostatic factors, and troponin T in CAPD patients. PMID- 12556075 TI - PD plus concept leads to significant increases in solute clearances in anuric CAPD patients. PMID- 12556077 TI - Sterile chemical peritonitis secondary to icodextrin: immunohistopathological description. PMID- 12556078 TI - Beneficial effect of icodextrin on the hypertriglyceridemia of CAPD patients. PMID- 12556079 TI - First report on the use of endoluminal FAS brush to restore flow in peritoneal catheters. PMID- 12556080 TI - High incidence of encapsulating peritoneal sclerosis in pediatric patients on peritoneal dialysis longer than 10 years. PMID- 12556081 TI - Pneumococcal and gonococcal peritonitis due to vaginitis. PMID- 12556082 TI - Gastric emptying in renal failure patients using the 13C-octanoic acid breath test: facts and artifacts. PMID- 12556083 TI - Is icodextrin peritonitis a reaction to bacterial endotoxins? PMID- 12556084 TI - Treatment with sevelamer decreases bicarbonate levels in peritoneal dialysis patients. PMID- 12556085 TI - No need to measure serum aluminum in patients starting chronic ambulatory peritoneal dialysis. PMID- 12556086 TI - Literature November-December 2002. PMID- 12556087 TI - How to improve cytologic screening for endocervical adenocarcinoma? AB - AIM: A retrospective study was undertaken to investigate how to improve the diagnosis of endocervical adenocarcinoma in screening programs. MATERIAL AND METHODS: The study group consisted of 29 slides of women diagnosed with cancer but who had negative smears. The slides were subdivided in 12 smears taken less than one year before diagnosis by histology and 17 smears taken between one and 10 years prior to diagnosis. A hundred smears of healthy women were used for comparison. All smears were studied macroscopically after which both groups of smears were scanned by the Neural Network Scanner (NNS). Differences between groups were studied for statistical significance using Pearson's Chi-squared test. FINDINGS: The macroscopic parameter of these smears found to be present most frequently was a heavy admixture of blood. The presence of blood (lysed or not) in the smears was equally consistently highlighted by the NNS. Statistical significance of the association of this parameter, with the presence of cancer, was demonstrated. CONCLUSION: The awareness of blood as a background feature of adenocarcinoma of the cervix will help to select cases needing special attention. These difficult bloody smears, studied by light microscopy and by NNS images can also be selected for additional MiB-1 staining. With this approach, blood in smears, otherwise frequently leading to a compromise of classification, can become a blessing in disguise. The diagnosis of endocervical adenocarcinoma in screening smears will therefore be improved. PMID- 12556088 TI - EUROGIN 2003 Conference. Facts, figures and impact. PMID- 12556089 TI - Evaluation of the circulating fraction of the HER-2/neu oncogene in patients with cervical cancer. AB - OBJECTIVE: To evaluate circulating HER-2/neu in cervical cancer patients prior to and following treatment. METHODS: Controls, and patients with either cervical dysplasia or cancer taken from an active gynecologic oncology service were evaluated for the expression of HER-2/neu in serum by ELISA before and following surgery, radiation, chemotherapy or combinations thereof. The resulant data was then evaluated for significance by either ANOVA or non-parametric testing. RESULTS: Mean differences were noted for patients with cervical cancer compared to controls. Patients with a good response to the chemotherapy indicated an increase in the serum oncogene, while those not responding either had no marked change or decreased the level of serum HER-2/neu. CONCLUSIONS: As serum HER-2/neu is a membrane bound portion of the intact molecule, these results suggest that due to the induction of cell death and breakdown, the liberation of this fraction (increased levels in the serum), is a viable indicator of response to treatment in some patients. A more detailed examination of this possibility along with expanded correlation with tissue expression is required. PMID- 12556090 TI - What is the role of interval blood testing in the management of chemotherapy for gynecologic malignancies. AB - PURPOSE: To determine the safety of omitting routine interval laboratory assessments, dietary restrictions, and isolation precautions between cycles of chemotherapy for gynecologic malignancies. METHODS: Data were retrospectively obtained from the records of women receiving chemotherapy for gynecologic cancer from July 1999-June 2000. Routine nadir determinations were not performed between treatment cycles; social interaction was encouraged, and pathogen-free diet recommendations were not provided. RESULTS: Eighty women received 449 cycles of chemotherapy. Four (5%) patients developed neutropenic fevers, and one of these women succumbed to sepsis. Eighteen (22.5%) women had 29 cycles delayed due to persistent myelosuppression when the ensuing chemotherapy infusion was to be administered. Hematopoietic growth factors overcame these delays during subsequent cycles in all but two patients. CONCLUSION: Omitting scheduled interval laboratory monitoring, dietary restrictions, and isolation precautions between chemotherapy cycles is convenient for patients, likely cost-effective, and does not increase morbidity in the gynecologic oncology population. PMID- 12556091 TI - Does FIGO clinical stage influence the survival of patients with early stages of uterine cervix carcinoma. International Federation of Gynaecology and Obstetrics. AB - We studied the influence of the FIGO clinical stage on the prognosis of 114 patients with early stages of uterine cervix carcinoma who had been treated with radical surgery in the "La Fe" Maternity Hospital in Valencia between 1971 and 1989. The prognosis became worse with each clinical stage, in line with the fact that the more advanced clinical stages are more likely to have certain types of spread, larger tumours and a greater stromal invasion depth. However, the Cox regression adjustment of the variables predicting disease-free and survival intervals, including the clinical stage and preoperative and postoperative treatment, did not reveal a significant link between clinical stage and the prognostic indices studied, while postoperative treatment variables showed a great predictive capacity, possibly due to the fact that the postoperative treatment used in more advanced stages is more aggressive. PMID- 12556092 TI - Prognostic significance of apoptotic index and bcl-2 and p53 expression in epithelial ovarian carcinoma. AB - PURPOSE OF THE STUDY: To determine if bcl-2 and p53 expression, and apoptotic index (AI) were associated with patient outcome in epithelial carcinomas of the ovary (EOC) and therefore useful as prognostic factors to predict survival. METHODS: A total of 50 women with epithelial carcinomas of the ovary were retrospectively analyzed. The archival paraffin-embedded material of of these cases were evaluated for expression of p53 and bcl-2 by immunohistochemical techniques. Apoptotic cells were detected with an in situ hybridisation method. RESULTS: A total of 33 (66%) of 50 cases showed positive immunoreactivity for the p53 antibody. Twenty-four of the 50 cases showed positive bcl-2 protein expression. Median value for AI was found to be 2.48. No statistically significant association was found between bcl-2 and p53 expression and clinicopathologic features. Univariate survival analysis of AI failed to reveal any effect on prognosis in the study population. CONCLUSION: We found neither p53 nor bcl-2 immunoreactivity to be of prognostic significance in patients with EOC. In addition, AI was not found to be an independent prognostic factor. PMID- 12556093 TI - Chlamydia trachomatis infection in women with CIN and invasive uterine cervix cancer. Significance of hormonal status. AB - In women with CIN at fertile age and those over 50 years of age, EGFR expression is lower in the presence of Chlamydia trachomatis (Cht) infection. In all Cht infected women over 50 years of age expression of Ki 67 is higher; the increase is significant among women with invasive carcinoma. In these groups of women with CIN and invasive carcinoma TGF-alpha expression is insignificantly augmented. Chronic Cht infection is associated with cervical hypertrophy. PMID- 12556094 TI - Sialyltransferase family members and cervix squamous cell carcinoma. AB - Sialic acids including a number of their derivatives are ubiquitous at the terminal positions of the oligosaccharides of glycoproteins. The transfer of sialic acids from cystidine-5-monophospho-N-acetylneuraminic acid (CMP-NeuAc) to the terminal position of the carbohydrate group of glycoproteins and glycolipids is catalyzed by a family of sialyltransferases (STs). There is a large body of evidence to suggest that tumor cells have altered surface properties from their normal counterparts, and that these changes are partially due to altered sialo glycoconjugates expressed on the plasma membrane and that altered sialylation (change in glycoprotein expression), which occurs during certain pathological processes, such as oncogenic transformation, tumor metastases, and invasion, is associated with enhanced ST activity. In this report we attempt to review the important findings in studing sialyltransferases of cervix squamous cell carcinoma. PMID- 12556095 TI - Vulvar lichen sclerosus in postmenopausal women: a comparative study for treating advanced disease with clobetasol propionate 0.05%. AB - BACKGROUND AND OBJECTIVE: Clobetasol propionate 0.05% has been the mainstay in treating vulvar lichen sclerosus (VLS) for the past ten years. The usual length of therapy is two to 12 weeks. We conducted this study to evaluate the efficacy and safety of treating severe lesions of VLS in postmenopausal women for a longer time on a regular basis using clobetasol propionate. MATERIALS AND METHODS: From 1997-2000, 137 women with VLS were examined in the Colposcopy and Laser Surgery Unit of "Alexandra" Hospital. Patients who were premenopausal, had previous therapy, exhibited mild or moderate disease or showed VIN or invasive cancer on vulvar biopsies were excluded from the study. The remaining women were divided into two groups. The first group applied clobetasol propionate 0.05% for three months and afterwards on an "as required" basis, whereas the second group used the ointment for six months on a regular basis. All patients were examined at two, three, six and 12 months following treatment. Signs and symptoms before and after therapy as well as side-effects caused by the ointment were recorded. RESULTS: The mean age of the women was 60.2 years. Fifty-four patients were divided into two categories. In the 6-month follow-up, 59% of the 1st group and 85% of the second had complete response regarding their symptoms whereas on the 12-month follow-up, the respective numbers were 48% and 74%. Concerning the signs, 30% of the first group and 55.5% of the second showed to have complete response after six months and 26% and 41% respectively after 12 months. All differences between the two groups, except the signs after 12 months, were statistically significant. There were no side-effects from the long-term use of clobetasol propionate 0.05%. CONCLUSIONS: Conservative management of severe lesions of VLS in postmenopausal women using clobetasol propionate 0.05% for a long time (6 months) on a regular basis, seems to be a safe and effective therapy. Improvement is observed primarily on the symptoms and less on the signs. PMID- 12556097 TI - Cancer pain, pathophysiology, characteristics and syndromes. AB - In this study the pathophysiology and characteristics of cancer pain together with cancer pain syndromes and guidelines of management are reviewed. Tumour associated pain may be nociceptive (somatic or visceral) if the sustaining mechanisms are related to ongoing tissue pathology, or neuropathic when pain is associated with injury to neural tissues. The mechanism by which tumours produce pain include obstruction of lymphatic and vascular channels, distension of a hollow viscous, oedema and tissue inflammation or necrosis. Injury to tissues results in the local release of numerous chemicals that mediate transmission of pain stimulus. Cancer pain syndromes result from one or more of three fundamental causes; direct tumour involvement of tissues, cancer-directed therapy, and mechanisms unrelated to cancer or its treatment. Cancer pain syndromes are also classified as acute or chronic. Cancer pain characteristics provide some of the data essential for syndrome identification. These characteristics include intensity, quality, distribution and temporal relationships. The principles of tumour-directed pain control include modifying the source of pain by treating the cancer and the inflammatory response to cancer, altering the central perception of pain and interfering with nociceptive transmission within the central nervous system. PMID- 12556096 TI - Depot leuprorelin acetate versus danazol in the treatment of infertile women with symptomatic endometriosis. AB - PURPOSE OF INVESTIGATION: Endometriosis is a common finding in women with infertility, but the mechanism by which it renders a woman infertile remains unclear. The medical treatment of pelvic endometriosis includes hormonal therapy that directly attacks endometriosis lesions or indirectly by inhibiting endometrial proliferation through estrogenic deprivation. The aim of this study was to compare the efficacy and safety of leuprorelin acetate depot and danazol for endometriosis in infertile women. METHODS: This randomized trial involved 81 women 19-41 years old with regular menses and known pelvic endometriosis who were recruited from the Fertility Center of the Second University of Naples between 1992 and 1999. Fifty-four women were given 3.75 mg of leuprolide acetate depot every 28 days for 24 weeks and the remaining 27 took 200 mg of danazol three times daily for 24 weeks. Efficacy assessments were based on pre-admission and end-of-treatment laparoscopic scores and subjective symptoms scores at 4-week intervals during and after treatment. Safety was evaluated by adverse events and clinical laboratory tests. RESULTS: In each group, endometriosis growth and symptoms significantly improved during treatment (p < 0.001). Significantly fewer patients randomized to leuprorelin acetate (5.5%) withdrew during treatment compared with 18.5% randomized to danazol (p < 0.05). After treatment symptoms returned in each group, but severity was less than at admission at all time points (p < 0.02). Hypoestrogenic side-effects were more common in those receiving leuprorelin, particularly hot flushes, but anabolic/androgenic side effects of weight gain and acne were more common in those receiving danazol. CONCLUSION: Both leuprorelin acetate depot and danazol are effective in the treatment of endometriosis in infertile patients. The hypoestrogenic side-effects of leuprorelin may be better tolerated than the androgenic, anabolic effects of danazol. PMID- 12556098 TI - Gains and losses of CD8, CD20 and CD56 expression in tumor stroma-infiltrating lymphocytes compared with tumor-associated lymphocytes from ascitic fluid and lymphocytes from tumor draining lymph nodes in serous papillary ovarian carcinoma patients. AB - Serous papillary ovarian cancer (SPC) is a highly aggressive tumor. About two thirds of women have advanced disease at the time of diagnosis. Although many women with disseminated disease respond at first to combinations of surgery and chemotherapy, nearly 90% of tumors recur and women die of disease. Update progress in our knowledge of tumor-associated antigens and insight into mechanisms involved in immune-mediated recognition of these antigens, have provided a strong starting point for using the immune system as a model for novel therapy. In this study we determined the immunological profile of tumor infiltrating lymphocytes (TILs), tumor-associated lymphocytes (TALs) in ascitic fluids, and lymphocytes from tumor draining regional lymph nodes (LNs) in SPC patients by CD20 (L26), CD8, and CD56 immunostaining. We examined 14 cases of TILs, 15 cases of TALs and 19 cases of LNs. TILs were infiltrating tumor stroma. No significant difference was detected in TILs, TALs and LNs in the expression of the B-cell marker CD20. In contrast, CD8 (T-cytotoxic) and CD56 (natural killer cell, NK) markers were dominant in LNs and TALs, but not in TILs. We conclude that SPC tumor lymphocytic infiltrate demonstrates a deplete T cytotoxic (CD8+) and NK cell (CD56+) immunophenotypic profile. This might in part explain the poor clinical outcome of the disease. PMID- 12556099 TI - Cytologic endometrial surveillance in tamoxifen-treated women. AB - The authors report the results of a study conducted on 136 patients affected by breast cancer under treatment with tamoxifen at a daily dose of 20 mg who underwent a strict follow-up including endometrial surveillance. The cytologic evaluation of the endometrium was performed on smears obtained by the endocyte sampling. The results were in accordance with what is reported in the literature. Forty cases presented with hyperplasia which was atypical in two cases. Only in one case did histology show a well differentiated adenocarcinoma. In 25 cases the endometrium resulted to be proliferative in accordance with the effect of tamoxifen, while the remaining 67 cases were atrophic. The investigation was not possible in four cases due to stenosis. In our study the endocyte sampler resulted to be an economic, simple and painless cytologic device, suitable for clinical use because of its low incidence of false positives. PMID- 12556100 TI - Prognostic significance of DNA topoisomerase II-alpha (Ki-S1) immunoexpression in endometrial carcinoma. AB - OBJECTIVE: In order to determine the significance of proliferative activity (PA) in endometrial carcinomas, we analysed the expression of cell cycle-related antigens in routinely processed tissue. MATERIALS AND METHODS: Serial sections of 113 endometrial carcinoma specimens were immunostained with the monoclonal antibody DNA Topoisomerase II-alpha (Ki-S1). In addition to Topoisomerase II alpha (Ki-S1) staining, histologic type, International Federation of Gynecology and Obstetrics (FIGO) stage. FIMO grade, depth of myometrial invasion, tumor size, lymphovascular space invasion, serosal and/or adnexal involvement, lymph node metastasis, age and peritoneal cytology were evaluated as prognostic indicators. The median follow-up time was 23 (range, 1 to 126 ) months. RESULTS: FIGO stage, FIGO grade, tumor size, lymphovascular space invasion, lymph node metastasis, peritoneal cytology and Topoisomerase II-alpha (Ki-S1) expression all significantly influenced survival in univariate analyses (p < or = 0.05). In the Cox regression analysis, Topoisomerase II-alpha (Ki-S1), serosal and/or adnexal involvement, and lymph node metastasis expression were the only variables with independent prognostic impact (p < or = 0.05), whereas FIGO stage, FIGO grade, histologic type FIGO grade, depth of invasion, tumor size, lymphovascular space invasion, age and peritoneal cytology had no independent influence (p > 0.05). Topoisomerase II-alpha (Ki-S1) staining was significantly elevated in advanced (Stage II, III, IV) as opposed to early (Stage I) carcinomas (p < or = 0.05). CONCLUSION: The association with established prognosticators for endometrial carcinomas, and the results of uni- and multivariate analysis indicate that the additional evaluation of DNA Topoisomerase II-alpha (Ki-S1) peptide antibody (PA) is useful for classifying patients into subgroups with low and high risk of relapse which might help to individualize the therapeutic strategy in endometrial carcinomas. PMID- 12556101 TI - Cervical cuff carcinoma: causes and prevention. AB - Over an 11-year period (1990-2000) a total of 59 patients with cervical cuff carcinoma were hospitalized (I stage--17; II stage--30; III-IV stage--12 patients). Seventeen patients were subjected to radical extirpation with lymphadenectomy for cervical cuff carcinoma, while 42 patients received combined radiotherapy. In the last 20-year period the patients had undergone supravaginal amputation of the uterus due to uterine fibromyoma, polyposis, and ovarian tumors in various gynecological clinics. In 12 patients the onset of cervical cuff carcinoma occurred within one year after supravaginal amputation of the uterus; in another six patients cancer developed within two years postoperatively, 15 patients in a 3 to 9 year period and 26 developed cancer 10-20 years after surgery. It seems in fact that when cervical cuff carcinoma developed within one year after supravaginal amputation of the uterus the patients had cervical carcinoma which was not identified due to physician negligence. In cases of latent periods up to ten years the patients must have had at least pre-existing disease. To prevent development of cervical cuff carcinoma in the future when operating on the uterus and/or adnexa the scope of surgery (extirpation, amputation) shall be decided based on colposcopic examination of the cervix (target cytology). PMID- 12556102 TI - p53 expression in tissue adjacent to endometrial carcinoma. AB - OBJECTIVE: Since several investigations did not demonstrate the presence of altered p53 in endometrial hyperplasias, it has been concluded that these alterations constitute a relatively late event in endometrial carcinoma. The aim of the present study was to assess the presence of p53 in the tissue adjacent to endometrial carcinoma in attempt to elucidate the relationship between these tissues. METHODS: New slides were prepared from paraffin-embeded tissue blocks of 49 endometrial endometrioid carcinoma hysterectomy specimens so that in each case tumor tissue and adjacent uninvolved endometrium were represented. Immunohistochemical staining for p53 detection was then performed. RESULTS: In 43 of the 49 hysterectomy specimens evaluated, the tissue adjacent to the endometrial carcinoma was non hyperplastic and in six it was hyperplastic. Positive immunohistochemical staining was found in 22 (44.9%) of endometrial carcinomas and in eight (16.3%) of the adjacent tissues. A statistically significant higher percentage of hyperplastic adjacent tissues than non hyperplastic adjacent tissues were immunohistochemically p53 positive (50.0% vs 11.3%; p = 0.047). CONCLUSIONS: Our findings may indicate that p53 alterations are not necessarily a late event in endometrial endometrioid carcinogenesis. Since a large proportion of tissues adjacent to endometrial carcinoma do not show p53 alterations, other early cellular events may also play a role. PMID- 12556103 TI - Vulvar Paget's disease: review of the literature, considerations about histogenetic hypothesis and surgical approaches. AB - Paget's disease of the vulva is a rare neoplasm that occurs on the apocrine glands. It predominantly is an intraepithelial lesion, but has the potential for dermal invasion and on occasion has been associated with an underlying adenocarcinoma. PMID- 12556104 TI - Surgical stage III endometrial cancer: analysis of treatment outcomes, prognostic factors and failure patterns. AB - OBJECTIVE: The purpose of this study was to evaluate the survival estimates of stage III endometrial cancer patients, and also to detect the prognostic factors and failure patterns. MATERIALS AND METHODS: Sixty-eight surgical Stage III endometrial cancer patients treated at Hacettepe University Hospital were included. All patients underwent surgical staging procedures consisting of peritoneal cytology, infracolic omentectomy, total abdominal hysterectomy, bilateral salpingo-oophorectomy and complete pelvic-paraaortic lymphadenectomy. By surgical staging 26 (38%) patients had Stage IIIA and 42 (62%) patients had Stage IIIC disease. The mean resected lymph node number was 26 (median, 25; range, 15-58). RESULTS: The median age was 60 years (range, 38-77), and the median follow-up period was 62 months (range, 36-90 months). The 5-year disease free survival rate was 58% and the 5-year overall survival rate was 64%. These figures for Stage IIIA were 60% and 68%, respectively; and for Stage IIIC they were 57% and 62%, respectively. No significant survival difference was detected between Stage IIIA and IIIC (p = 0.60 for disease-free survival and p = 0.48 for overall survival). High grade and positive peritoneal cytology predicted poor survival in both univariate (p = 0.004 and p = 0.006, respectively) and multivariate (p = 0.05 and p = 0.04, respectively) analysis. Twenty-eight patients (41%) had recurrence with a median time of 23 months (range, 10-54 months). Nine patients (13%) had only local, 13 patients (19%) had only distant and six patients (9%) had both local and distant relapse. CONCLUSION: Surgical staging is important in the management of Stage III endometrial cancer, and the main problem is still distant failure. In multivariate analysis high grade and positive peritoneal cytology predicted poor survival significantly. PMID- 12556105 TI - Effects of tamoxifen administration in rat vaginas: an ultrastructural and light microscopy study. AB - BACKGROUND: Tamoxifen (TAM) inhibits the initiation of carcinogen induced rat mammary tumours and is administrered for extended periods after the initiation of carcinogenesis. It is also a widely used treatment for breast and gynaecological cancer. OBJECTIVE: In this study, we aimed to investigate tamoxifen (TAM) administration on vagen development in rats. MATERIAL & METHODS: Twenty sexually mature and pregnant Wistar albino rats were chosen as the animal model. They were divided into two groups. Group I: Control group, Group II: Tamoxifen applied (between gestational day 16 and 21 days); 100 microg tamoxifen citrate (TAM) in 0.05 ml saline subcutaneously per day/animal. After birth, all female rats were sacrificed on the 60th day and were taken vaginal tissue. Transmission electron microscopy and light microscopy have been used to study changes to the vaginal epithelium. RESULTS: A statistically significant reduction of birth body weight was noted in the experimental group of rats when compared to the control group (P < 0.05). We saw increase in the thickness of the epithelium layer and irregularity and disappearance of microscopic papilla, cytoplasmic vacuolation in cells of the surface layer, thin and irregular basal membrane, lateral junction of cells were destroyed in the TAM treated groups. In conclusion, neonatal tamoxifen administration affects vagina epithelium and lead to decreasing birth body weight and vaginal adenosis. PMID- 12556106 TI - Abdominal wall metastasis from ovarian cancer after laparotomy. A case report. AB - A 27-year-old woman underwent surgery for an abdominal wall mass later confirmed to be a relapse of Stage I ovarian mucinous adenocarcinoma. The authors stress that caution should be observed during laparotomy to remove a malignant neoplasm in order to avoid parietal dissemination. PMID- 12556107 TI - Endometrial metastasis from signet-ring breast carcinoma: case report. AB - Breast cancer rarely metastasizes to the endometrium. The signet-ring variety of breast cancer is very rare and has been reported to metastasize to the endometrium in only three previous reports. We present a patient with signet-ring breast carcinoma with metastasis to the endometrium. The patient presented with vaginal bleeding and was found to have wide spread metastasis. Endometrial metastasis in patients with signet-ring breast cancer usually denotes a poor prognosis. PMID- 12556108 TI - Intraperitoneal haemorrhage secondary to perforation of uterine fibroid after cystic degeneration. Unusual CT findings resembling malignant pelvic tumor: case report. AB - Intraperitoneal haemorrhage is a rare complication of myomatous uterus. We present a case of a 37-year-old white nullipara who presented in the emergency room with acute, lower-abdominal pain which reportedly started after riding over a bump on a motorcycle. On examination, the abdomen was diffusely tender, with moderate spasm and rebound tenderness in both iliac fossae. Pregnancy test was negative. Computed tomography revealed a soft-tissue mass with cystic components and inhomogeneous appearance. Free fluid in the peritoneal cavity suggested ascites. The patient underwent an exploratory laparotomy. A ruptured, actively bleeding, subserosal, nonpedunculated, cystic degenerated uterine fibroid was found, as well as approximately two liters of free, bloodstained peritoneal fluid and clots. Subtotal hysterectomy without salpingo-oophorectomy was performed, followed by evacuation of the fluid and clots. The patient's postoperative course was uneventful. In conclusion, definitive, preoperative diagnosis of a perforated, haemorrhaging, uterine fibroid is difficult; exploratory laparotomy is both diagnostic and therapeutic in this rare, life-threatening condition. PMID- 12556109 TI - Ultrasonographic and computed tomography manifestations of intussusception secondary to primary non-Hodgkin's lymphoma diagnosed in puerperium: report of a case and review of the literature. AB - Non-Hodgkin's lymphoma is infrequently diagnosed during pregnancy and puerperium. A 21-year-old woman, para II, gravida II, on the seventh postpartum day arrived at the Emergency Gynaecologic Clinic complaining of colicky abdominal pain associated with bilious vomiting. A large tender mass of firm consistency was palpated in the midline towards the left hypochondrium. A CT scan of the upper abdomen showed thickened small bowel with halo, specifically at the junction of the jejunum with the ileum, resembling imaging of intussusception. Also, blocks of mesenteric and paraortic lymph nodes and multiple lymph nodes inducing enlargement of the anterior mediastinum were found. These findings rendered a picture of lymphoma in Stage at least III. Subsequent exploratory laparotomy confirmed our preoperative diagnosis. Histopathological examination of the lymph node biopsies established the diagnosis of a primary non-Hodgkin's lymphoma. Judging from the extent of the non-Hodgkin's lymphoma in our patient seven days after delivery the disease probably already existed during the last trimester of pregnancy. This study demonstrates a remarkable delay in diagnosis of non Hodgkin's lymphoma in pregnancy. The diagnostic dilemmas of non-Hodgkin's lymphoma during pregnancy are emphasized and the literature is reviewed. PMID- 12556110 TI - Primary invasive vaginal cancer. Report of 12 cases. AB - The aim of this study was to describe the clinical features, methods of treatment and results of treatment among patients with primary invasive cancer. Twelve women in whom primary invasive cancer of the vagina was diagnosed between 1996 and 2001 were studied. The mean age of women with primary invasive vaginal cancer was 70.1 years. Half of the patients had advanced stage carcinoma at the time of the diagnosis and 85% of patients had not had a Pap-smear in the last 15 years. Treatment consisted of radiotherapy or a combination of surgery and radiotherapy. Four patients out of six with advanced stage disease died in 30 months. In conclusion, the present study confirms that early detection of the disease could lead to more successful management and therefore better prognosis. PMID- 12556111 TI - Vaginal fibroma. Case report. AB - A case of a vaginal fibroma in a 50-year-old woman is presented. The tumor presented as a solid mass in the upper third of the anterior vaginal wall. Intravenous pyelography (IVP) and cystoscopy excluded any association of the mass with the urinary tract. Surgical excision of the tumor was easily accomplished and histopathological examination established the diagnosis of a fibroid tumor of the vagina. PMID- 12556112 TI - Primary small-cell carcinoma of the endometrium: clinicopathological study of a case and review of the literature. AB - BACKGROUND: Small-cell carcinomas are almost always primary in the lungs and are highly malignant. These tumors may also occur in the female genital tract. However, primary small-cell carcinoma of the endometrium is extremely rare with very few cases reported in the English literature. This tumor may exhibit evidence of neuroendocrine differentiation and has a high propensity for systemic spread and poor prognosis. CASE: A 55-year-old postmenopausal woman with primary small-cell carcinoma of the endometrium, FIGO stage Ib, underwent total abdominal hysterectomy with bilateral salpingo-oophorectomy and sampling node biopsies of the parametrial spaces, followed by adjuvant combined chemotherapy. CONCLUSION: A case of small-cell carcinoma of the endometrium, is reported and its clinical, histological and immunohistochemical features are discussed. PMID- 12556113 TI - Shyness and anxiety as predictors of patterns of Internet usage. AB - The Internet has the potential to empower or isolate. Shyness and anxiety may potentially influence the extent to which people avail themselves of Internet services such as email, chat rooms, information searches, entertainment, and commerce. To understand how personality moderates Internet usage, 177 participants completed an Internet Use Survey, the Social Reticence Scale, and a Trait Anxiety Inventory. Shyness, anxiety, gender, and academic achievement were employed within separate multiple regressions to predict forms of Internet usage. The use of email and chat-rooms was not related to shyness or anxiety, suggesting that shyness or anxiety does not pose an obstacle to these Internet applications. Males were more likely to use the Internet for downloading entertainment. Shy males were more likely to use the Internet for recreation/leisure searches. Highly educated males were more likely to use the Internet for banking and paying bills. Although shyness or anxiety does not seem to modify the communicative functions of the Internet, it may influence people's use of other recreational applications. PMID- 12556114 TI - Users divided? Exploring the gender gap in Internet use. AB - Many believe that males and females use and regard computer technology differently. Males are generally assumed to be more comfortable with, more adaptable to, and less anxious with computer technology. The same biases are now being applied specifically to Internet technology. Based on research showing that men prefer to use the Internet for information gathering and entertainment, while women prefer to use the Internet for interpersonal communication, this study examined the effects of Internet use when both males and females engaged in the same activity. Participants engaged in synchronous, dyadic chat sessions, and changes in repeated measures of loneliness, depression, self-esteem, and perceived social support were tracked over time. Although previous studies have concluded not only that males and females differ in their computer cognitions and attitudes, but also that they differ in the types of applications they pursue online, no gender differences were found in the present study. PMID- 12556115 TI - Virtual reality exposure therapy for World Trade Center Post-traumatic Stress Disorder: a case report. AB - Done properly by experienced therapists, re-exposure to memories of traumatic events via imaginal exposure therapy can lead to a reduction of Post-traumatic Stress Disorder (PTSD) symptoms. Exposure helps the patient process and habituate to memories and strong emotions associated with the traumatic event: memories and emotions they have been carefully avoiding. But many patients are unwilling or unable to self-generate and re-experience painful emotional images. The present case study describes the treatment of a survivor of the World Trade Center (WTC) attack of 9-11-01 who had developed acute PTSD. After she failed to improve with traditional imaginal exposure therapy, we sought to increase emotional engagement and treatment success using virtual reality (VR) exposure therapy. Over the course of six 1-h VR exposure therapy sessions, we gradually and systematically exposed the PTSD patient to virtual planes flying over the World Trade Center, jets crashing into the World Trade Center with animated explosions and sound effects, virtual people jumping to their deaths from the burning buildings, towers collapsing, and dust clouds. VR graded exposure therapy was successful for reducing acute PTSD symptoms. Depression and PTSD symptoms as measured by the Beck Depression Inventory and the Clinician Administered PTSD Scale indicated a large (83%) reduction in depression, and large (90%) reduction in PTSD symptoms after completing VR exposure therapy. Although case reports are scientifically inconclusive by nature, these strong preliminary results suggest that VR exposure therapy is a promising new medium for treating acute PTSD. This study may be examined in more detail at www.vrpain.com. PMID- 12556116 TI - Study of Internet use by customers of a large French store for computers. AB - The objective of this work was to study the relation to the Internet of subjects who own a computer and who use it for professional or personal reasons. We interviewed 150 customers of the biggest French store that specializes in computers. Each day, the mean of duration for connecting time was 6.2 h (SD = 4). Customers were connected 3.7 h (SD = 3.3) between 8:00 a.m. and 6:00 p.m., 1.8 h (SD = 1.6) between 6:00 p.m. and 11:00 p.m., and 0.5 hours at night (SD = 1.3). Men were more often connected between 6:00 p.m. and 11:00 p.m. We found a negative correlation between age and duration of connection to internet. Age was negatively correlated to the time of connection between 6:00 p.m. and 11:00 p.m. and between 11:00 a.m. and 8:00 a.m. The number of days of holidays was negatively correlated to total time of connection (p < 0.001) and to connection between 6:00 p.m. and 11:00 p.m. (p = 0.04). No correlation was found between the time spent watching television or speaking on a cellular phone and parameters of Internet use. High consumers of games are connected 3.4 h every evening and 1.2 h each night. They spend, in addition, 30 min each day connected to virtual casinos. The typical high consumer of the Internet is young and interested in games on-line, he takes fewer days of holiday than others, and he sends and receives more e-mails than other computer users. PMID- 12556117 TI - Brief virtual reality therapy for public speaking anxiety. AB - The primary goal of this research program was to investigate the effectiveness of virtual reality therapy (VRT) in reducing public speaking anxiety of university students. The prevalence and impact of public speaking anxiety as a type of Social Phobia are discussed. Studies of VRT as an emerging treatment for psychological problems are reviewed. In the present study, eight students completed VRT individual treatment and post-testing, and six students in a Wait List control group completed post-testing. Assessment measures included four self report inventories, self-report of Subjective Units of Discomfort during exposure to VRT and physiological measurements of heart rate during speaking tasks. Four weekly individual exposure treatment sessions of approximately 15 min each were conducted by the author serving as therapist. Results on self-report and physiological measures appear to indicate that four virtual reality treatment sessions were effective in reducing public speaking anxiety in university students, corroborating earlier studies of VRT's effectiveness as a psychotherapeutic modality. Future research directions are discussed, primarily the need for research on younger populations, to assess the effectiveness of VRT for earlier intervention with public speaking anxiety. PMID- 12556118 TI - College students' Internet use, relationship formation, and personality correlates. AB - The Internet has become a standard fixture in the lives of many people, with communication being one of its most popular uses. Several mediums-such as electronic mail (e-mail), group mailing forums, interactive games, and real-time chatting-provide users with communication opportunities. Through extended communication on the Internet, many users have formed relationships with others online. Several aspects of the Internet medium interact to make the course of relationship development online differ from offline development. The Internet is still a rather uncharted area in terms of psychological research, especially in the field of online relationship development. This research examined college students' Internet use, relationship formation, and personality characteristics. No differences were found between participants who did and did not form new relationships online. However, differences were present within the group that formed online relationships, such as ease at finding similar others online as well as differences in social skills and loneliness scores. This calls for a reexamination of previous hypotheses regarding individuals who form relationships online. PMID- 12556119 TI - Virtual reality and the person-environment experience. AB - This paper discusses how a projected virtual reality system that uses video gesture recognition technology can influence the person-environment process. The psychosocial concepts of embodiment, environmental centralization, and environmental personalization are illustrated with examples of virtual reality applications with children with disabilities. Through these, the advantages of using virtual reality to influence the person-environment relationship are discussed. Disadvantages of implementing this virtual reality approach are also presented as well as recommendations for future work in this area. PMID- 12556120 TI - An evaluation model for psychoeducational interventions using interactive multimedia. AB - A review of published evaluations of psychoeducational interventions using interactive digital multimedia shows that evaluations typically address only some of the areas that collectively would constitute a comprehensive evaluation. There appears to be a need for an accepted standard for these evaluations, based on a general evaluation model that encompasses all relevant aspects of development, efficacy and use of multimedia interventions. A comprehensive model is proposed which includes special features of multimedia interventions that lead to unique evaluation requirements. The model integrates relevant aspects of program evaluation and clinical trials models in order to provide a unique model that includes all the evaluation domains relevant to digital multimedia interventions. These include evaluation of intervention theory, intervention design strategies, the formative process, intervention efficacy (process and outcome) and contextual issues such as evaluability assessment, reporting and stakeholder issues. The application of individual components of the model is illustrated with reference to problems in the evaluation literature on a particular type of digital multimedia intervention, electroencephalographic biofeedback for Attention Deficit/Hyperactivity Disorder. The model should be useful for researchers planning evaluations of digital multimedia interventions, especially in the psychoeducational domain. This paper provides a theoretical and evidential background for the evaluation model, and includes a checklist and flowchart for the planning and conduct of the evaluation. PMID- 12556121 TI - The sociocognitive psychology of computer-mediated communication: the present and future of technology-based interactions. AB - The increased diffusion of the Internet has made computer-mediated communication (CMC) very popular. However, a difficult question arises for psychologists and communication researchers: "What are the communicative characteristics of CMC?" According to the "cues-filtered-out" approach, CMC lacks the specifically relational features (social cues), which enable the interlocutors to identify correctly the kind of interpersonal situations they find themselves in. This paper counters this vision by integrating in its theoretical frame the different psycho-social approaches available in current literature. In particular, the paper describes the characteristics of the socio-cognitive processes-emotional expression, context definition, and identity creation-used by the interlocutors to make order and create relationships out of the miscommunication processes typical of CMC. Moreover, it presents the emerging forms of CMC-instant messaging, shared hypermedia, weblogs, and graphical chats-and their possible social and communicative effects. PMID- 12556122 TI - Strategies used by rhizobia to lower plant ethylene levels and increase nodulation. AB - Agriculture depends heavily on biologically fixed nitrogen from the symbiotic association between rhizobia and plants. Molecular nitrogen is fixed by differentiated forms of rhizobia in nodules located on plant roots. The phytohormone, ethylene, acts as a negative factor in the nodulation process. Recent discoveries suggest several strategies used by rhizobia to reduce the amount of ethylene synthesized by their legume symbionts, decreasing the negative effect of ethylene on nodulation. At least one strain of rhizobia produces rhizobitoxine, an inhibitor of ethylene synthesis. Active 1-aminocyclopropane-1 carboxylate (ACC) deaminase has been detected in a number of other rhizobial strains. This enzyme catalyzes the cleavage of ACC to alpha-ketobutyrate and ammonia. It has been shown that the inhibitory effect of ethylene on plant root elongation can be reduced by the activity of ACC deaminase. PMID- 12556123 TI - Soil and plant effects on microbial community structure. AB - We investigated the effects of two different plant species (corn and soybean) and three different soil types on microbial community structure in the rhizosphere. Our working hypothesis was that the rhizosphere effect would be strongest on fast growing aerobic heterotrophs, while there would be little or no rhizosphere effect on oligotrophic and other slow-growing microorganisms. Culturable bacteria and fungi had larger population densities in the rhizosphere than in bulk soil. Communities were characterized by soil fatty acid analysis and by substrate utilization assays for bacteria and fungi. Fatty acid analysis revealed a very strong soil effect but little plant effect on the microbial community, indicating that the overall microbial community structure was not affected by the rhizosphere. There was a strong rhizosphere effect detected by the substrate utilization assay for fast-growing aerobic heterotrophic bacterial community structure, with soil controls and rhizosphere samples clearly distinguished from each other. There was a much weaker rhizosphere effect on fungal communities than on bacterial communities as measured by the substrate utilization assays. At this coarse level of community analysis, the rhizosphere microbial community was impacted most by soil effects, and the rhizosphere only affected a small portion of the total bacteria. PMID- 12556124 TI - Increasing fermentation efficiency at high sugar concentrations by supplementing an additional source of nitrogen during the exponential phase of the tequila fermentation process. AB - In the tequila industry, fermentation is traditionally achieved at sugar concentrations ranging from 50 to 100 g x L(-1). In this work, the behaviour of the Saccharomyces cerevisiae yeast (isolated from the juices of the Agave tequilana Weber blue variety) during the agave juice fermentation is compared at different sugar concentrations to determine if it is feasible for the industry to run fermentation at higher sugar concentrations. Fermentation efficiency is shown to be higher (above 90%) at a high concentration of initial sugar (170 g x L(-1)) when an additional source of nitrogen (a mixture of amino acids and ammonium sulphate, different than a grape must nitrogen composition) is added during the exponential growth phase. PMID- 12556125 TI - Effect of fusaric acid and phytoanticipins on growth of rhizobacteria and Fusarium oxysporum. AB - Suppression of soilborne diseases by biocontrol agents involves complex interactions among biocontrol agents and the pathogen and between these microorganisms and the plant. In general, these interactions are not well characterized. In this work, we studied (i) the diversity among strains of fluorescent Pseudomonas spp., Bacillus spp., and Paenibacillus sp. for their sensitivity to fusaric acid (FAc) and phytoanticipins from different host plants, (ii) the diversity of pathogenic and nonpathogenic Fusarium oxysporum isolates for their sensitivity to phytoanticipins, and (iii) the influence of FAc on the production of pyoverdine by fluorescent Pseudomonas spp. tolerant to this compound. There was a great diversity in the response of the bacterial strains to FAc; however, as a group, Bacillus spp. and Paenibacillus macerans were much more sensitive to FAc than Pseudomonas spp. FAc also affected production of pyoverdine by FAc-tolerant Pseudomonas spp. strains. Phytoanticipins differed in their effects on microbial growth, and sensitivity to a phytoanticipin varied among bacterial and fungal strains. Biochanin A did not affect growth of bacteria, but coumarin inhibited growth of Pseudomonas spp. strains and had no effect on Bacillus circulans and P. macerans. Conversely, tomatine inhibited growth of B. circulans and P. macerans. Biochanin A and tomatine inhibited growth of three pathogenic isolates of F. oxysporum but increased growth of three nonpathogenic F. oxysporum isolates. Coumarin inhibited growth of all pathogenic and nonpathogenic F. oxysporum isolates. These results are indicative of the complex interactions that can occur among plants, pathogens, and biological control agents in the rhizosphere and on the root surface. Also, these results may help to explain the low efficacy of some combinations of biocontrol agents, as well as the inconsistency in achieving disease suppression under field conditions. PMID- 12556126 TI - The pathway of dephosphorylation of myo-inositol hexakisphosphate by phytate degrading enzymes of different Bacillus spp. AB - The pathway of dephosphorylation of myo-inositol hexakisphosphate by the phytate degrading enzymes of Bacillus subtilis 168, Bacillus amyloliquefaciens ATCC 15841, and Bacillus amyloliquefaciens 45 was established using a combination of high-performance ion chromatography analysis and kinetic studies. The data demonstrate that all the Bacillus phytate-degrading enzymes under investigation dephosphorylate myo-inositol hexakisphosphate by sequential removal of phosphate groups via two independent routes; the routes proceed via D-Ins(1,2,4,5,6)P5 to Ins(2,4,5,6)P4 to finally Ins(2,4,6)P3 or D-Ins(2,5,6)P3 and via D Ins(1,2,4,5,6)P5 to D-Ins(1,2,5,6)P4 to finally D-Ins(1,2,6)P3. The resulting myo inositol trisphosphate D-Ins(1,2,6)P3 was degraded via D-Ins(2,6)P2 to finally Ins(2)P after prolonged incubation times in combination with increased enzyme concentration. PMID- 12556127 TI - Campylobacter fetus adheres to and enters INT 407 cells. AB - Campylobacter fetus is a Gram-negative bacterial pathogen of humans and ungulates and is normally transmitted via ingestion of contaminated food or water with infection resulting in mild to severe enteritis. However, despite clinical evidence that C. fetus infection often involves transient bacteremic states from which systemic infection may develop and the frequent isolation of C. fetus from extra-intestinal sites, this organism displays very poor invasiveness in in vitro models of infection. In this study, immunofluorescence microscopy and gentamicin protection assays were used to investigate the ability of six clinical isolates and one reference strain of C. fetus to adhere to and invade the human intestinal epithelial cell line, INT 407. During an initial 4-h infection period, all C. fetus strains were detected intracellularly using both techniques, though adherence and internalization levels were very low when determined from gentamicin protection assays. Microscopy results indicated that during a 4-h infection period, four of the five clinical strains tested were adherent to 41.3 87.3% of INT 407 cells observed and that 25.2-34.6% of INT 407 cells contained intracellular C. fetus. The C. fetus reference strain displayed the lowest levels of adherence and internalization. A modified infection assay revealed that C. fetus adherence did not necessarily culminate in internalization. Despite the large percentage of INT 407 cells with adherent bacteria, the percentage of INT 407 cells with intracellular bacteria remained unchanged when incubation was extended from 4 h to 20 h. However, microscopy of INT 407 cells 24 h postinfection (p.i.) revealed that infected host cells contained clusters of densely packed C. fetus cells. Gentamicin protection assays revealed that intracellular C. fetus cells were not only viable 24 h p.i. but also that C. fetus had increased in number approximately three- to fourfold between 4 and 24 h p.i., indicative of intracellular replication. Investigation of the role of the host cell cytoskeleton revealed that pretreatment of host cells with cytochalasin D, colchicine, vinblastine, taxol, or dimethyl sulfoxide (DMSO) did not impact upon C. fetus adherence or internalization of INT 407 cells. Microscopy indicated neither rearrangement nor colocalization of either microtubules or microfilaments in INT 407 cells in response to C. fetus adherence or internalization. Together, these data indicate that clinical isolates of C. fetus are capable of adhering, entering, and surviving within the nonphagocytic epithelial cell line, INT 407. PMID- 12556128 TI - Characterization of ndvD, the third gene involved in the synthesis of cyclic beta (1 --> 3),(1 --> 6)-D-glucans in Bradyrhizobium japonicum. AB - Previously, we identified two genes in Bradyrhizobium japonicum (ndvB, ndvC) that are required for cyclic beta-(1 --> 3),(1 --> 6)-D-glucan synthesis and successful symbiotic interaction with soybean (Glycine max). In this study, we report a new open reading frame (ORF1) located in the intergenic region between ndvB and ndvC, which is essential for beta-glucan synthesis and effective nodulation of G. max. This new gene is designated ndvD (nodule development). The ndvD translation product has a predicted molecular mass of 26.4 kDa with one transmembrane domain. Genetic experiments involving gene deletion, Tn5 insertion, and gene complementation revealed that the mutation of ndvD generated pleiotropic phenotypes, including hypoosmotic sensitivity, reduced motility, and defects in conjugative gene transfer, in addition to symbiotic ineffectiveness. Although deficient in in vivo beta-glucan synthesis, membrane preparations from the ndvD mutant synthesized neutral beta-glucans in vitro. Therefore, ndvD does not appear to be a structural gene for beta-glucan synthesis. Our hypothesis for the mechanism of beta-(1 --> 3),(1 --> 6)-D-glucan synthesis is presented. PMID- 12556129 TI - Toxigenic diversity of two different RAPD groups of Stachybotrys chartarum isolates analyzed by potential for trichothecene production and for boar sperm cell motility inhibition. AB - Thirty-one isolates of Stachybotrys chartarum from indoor and outdoor environments were analyzed for the presence of the trichodiene synthase (Tri5) gene, trichothecenes, boar sperm cell motility inhibition, and randomly amplified polymorphic DNA banding patterns (RAPDs). Twenty-two S. chartarum isolates tested positive for the Tri5 gene and nine were negative when tested using novel Tri5 gene-specific PCR primer pair. The Tri5 gene positive isolates contained satratoxins (five isolates) or the simple trichothecene, trichodermol (11 isolates). The Tri5 gene negative isolates did not produce satratoxins or trichodermol. Nineteen S. chartarum isolates, distributed among the Tri5 gene negative and positive groups, inhibited boar spermatozoan motility at concentrations of < or = 60 microg of crude cell extract/mL. The inhibition of motility was independent of satratoxins or atranones. Unweighted pair group method of arithmetic averages (UPGMA) cluster analysis of RAPD fragments clustered the 31 S. chartarum isolates in two distinct groups designated as RAPD groups 1 and 2. The grouping of S. chartarum isolates obtained by UPGMA cluster analysis of RAPD fragments was identical to the grouping obtained by Tri5 gene specific PCR. This indicates that the S. chartarum isolates belonging to different groups were genetically distinct in a much wider area than just the Tri5 gene. PMID- 12556130 TI - Properties of a hydrophobin isolated from the mycoparasitic fungus Verticillium fungicola. AB - Verticillium fungicola, isolated from Agaricus bisporus (commercial mushroom), produced significant extracellular hydrophobin when grown for 7 days in a static liquid culture of synthetic minimal medium. The hydrophobin was purified by precipitation with ammonium sulphate (80% saturation), Sephadex G-100 gel filtration, and hydroxyapatite column chromatography. The purified protein yielded a single band in polyacrylamide gel electrophoresis under native conditions, with an apparent molecular mass of 70 +/- 4 kDa, and also another single band in SDS-PAGE, with a molecular mass of 7 +/- 3 kDa. Molecular mass determined with matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) resulted in 7563.9 m/z. The same protein was extracted from the V. fungicola mycelium. Analysis of the amino acid composition revealed the presence of about 50% hydrophobic residues, detecting at least six cysteines, evaluated as cystines, and no free sulfhydryl groups. The protein did not show any glycosylation. On the basis of similarities in hydropathy patterns and solubility characteristics, V. fungicola hydrophobin can be included as a new member of Class II hydrophobins. PMID- 12556131 TI - Expression of a glycogen synthase protein kinase homolog from Colletotrichum gloeosporioides f.sp. malvae during infection of Malva pusilla. AB - The potential role of a GSK3 protein kinase homolog, cggsk, was examined from Colletotrichum gloeosporioides f.sp. malvae, a fungal pathogen of Malva pusilla. A peak in cggsk expression relative to a constitutively expressed fungal actin gene occurred during host penetration and was followed by much lower expression levels during subsequent biotrophic and necrotrophic growth in host tissue. The peak level of cggsk expression observed during penetration was 21-fold greater than that during necrotrophic growth. Expression of cggsk showed small but reproducible changes during growth in culture; however, the levels were always similar to that during necrotrophic growth in the host. One possible role for cggsk could be to coordinate fungal development during host penetration. PMID- 12556132 TI - To pee or not to pee: reconsidering the need for urinalysis. AB - While urinalysis is used regularly as a tool to validate self-reported recent drug use, past research has been inconclusive in evaluating concordance between the two measures. In the current study, urinalysis results for cocaine and opiates are compared to self-reported three-day cocaine and opiate use with data collected through Houston's Arrestee Drug Abuse Monitoring (ADAM) Program between 1990 and 1999. Separate analyses are conducted for each year during the decade. Kappa statistics indicate that the strength of agreement between the two drug use measures is consistent over time. These findings suggest that the need for urinalysis should be reconsidered. PMID- 12556133 TI - Response distortion in adolescents who smoke: a pilot study. AB - A wide range of data are obtained with self-report. Information obtained from persons using substances is generally reliable and valid, however, many studies show that some proportion of self-reports regarding substance use are inaccurate. This study examines self-reported response distortion in adolescents who received a brief intervention to reduce their smoking. Findings indicate that age and ethnicity of respondent may influence reported response distortion. Factors that appear to influence under- and over-reporting include social desirability and fear of repercussion. Response distortion does not appear to affect rated usefulness of the intervention, nor does intervention type appear to influence whether respondents thought the researcher wanted them to report less use. Results point to the need for further research regarding adolescents and response distortion. PMID- 12556134 TI - The reliability of environmental measures of the college alcohol environment. AB - Much of what we know about students' drinking patterns and problems related to alcohol use is based on survey research. Although local and national survey data are important to alcohol-prevention projects, they do not sufficiently capture the complexity of the alcohol environment. Environmental prevention approaches to alcohol-related problems have been shown to be effective in community settings and researchers have begun to study and adapt such approaches for use on college campuses. Many environmental approaches require systematic scanning of the campus alcohol environment. This study assessed the inter-rater reliability of two environmental scanning tools (a newspaper content analysis form and a bulletin analysis form) designed to identify alcohol-related advertisements targeting college students. Inter-rater reliability for these forms varied across different rating categories and ranged from poor to excellent. Suggestions for future research are addressed. PMID- 12556135 TI - Tobacco use and quit behaviors among delinquent youth: a pilot study. AB - With rates of adolescent tobacco use steadily increasing over the past 20 years, assisting youth to quit, particularly those youth most susceptible to tobacco use, has become a national focus. The purpose of this pilot study was to examine and profile the smoking patterns of a small sample (n = 37) of juvenile offenders in Texas. In contrast to anecdotal reports, the sample group from this study did not differ clinically from the general population of adolescent tobacco users. Importantly, the study participants indicated previous attempts to stop smoking. The authors conclude that delinquent youth may be responsive to cognitive behavioral smoking cessation programs that also address the management of peer networks. PMID- 12556136 TI - How beliefs about substance use differ by socio-demographic characteristics, individual experiences, and neighborhood environments among urban adolescents. AB - In this article, we tested a series of Item Response Theory (IRT) models to examine the individual and neighborhood variation in perceived risk along dimensions of substance use (alcohol, marijuana, and hard drugs) and usage patterns (light/experimental use, moderate use, heavy/regular use). Data were gathered from 2266 adolescents aged 9, 12, and 15 residing in 79 Chicago neighborhoods. Developmental patterns for age and amount of use were observed whereby older respondents rated alcohol and marijuana as less harmful compared to the younger respondents, but rated hard drugs as more harmful. Risk perceptions were found to be more closely tied to one's direct experience with drugs rather than a general constellation of beliefs. Neighborhood variation in risk perceptions was also observed for hard drugs and three patterns of use, controlling for characteristics of individual residents. Neighborhoods did not vary in risk perceptions toward alcohol use. Individual-level factors rather than characteristics of the neighborhoods explained the observed neighborhood variation in perceptions toward marijuana use. These findings illustrate the complex links between individual and contextual factors in the development of beliefs about the health risks associated with substance use. PMID- 12556137 TI - Aspects of guilt and self-reported substance use in adolescence. AB - The use of addictive substances is undergoing moralization in American society behaviors once viewed as personal preferences now carry moral significance. Research has shown that sociomoral emotions like guilt, thought to be reflective of one's internalized standards and societal mores, can be an important influence on behavior. The present study explored the relationship between college students' self-reports of adolescent substance use (cigarettes, alcohol, and marijuana) and scores on indices tapping different aspects of guilt (Standards, Situational, and Chronic Guilt). Participants were 230 undergraduate students (mean age = 19; 55 percent female; 69 percent White). Substance users had lower scores on Standards and Situational Guilt than non-users, but no difference was observed in Chronic Guilt. The present results suggest that a stronger internalization of societal standards, as reflected by higher scores on Standards and Situational Guilt, may prove a useful tool in the prevention of substance use. PMID- 12556138 TI - Principles that underpin effective school-based drug education. AB - This study identifies the conceptual underpinnings of effective school-based drug education practice in light of contemporary research evidence and the practical experience of a broad range of drug education stakeholders. The research involved a review of the literature, a national survey of 210 Australian teachers and others involved in drug education, and structured interviews with 22 key Australian drug education policy stakeholders. The findings from this research have been distilled and presented as a list of 16 principles that underpin effective drug education. In broad terms, drug education should be evidence based, developmentally appropriate, sequential, and contextual. Programs should be initiated before drug use commences. Strategies should be linked to goals and should incorporate harm minimization. Teaching should be interactive and use peer leaders. The role of the classroom teacher is central. Certain program content is important, as is social and resistance skills training. Community values, the social context of use, and the nature of drug harm have to be addressed. Coverage needs to be adequate and supported by follow-up. It is envisaged that these principles will provide all those involved in the drug education field with a set of up-to-date, research-based guidelines against which to reference decisions on program design, selection, implementation, and evaluation. PMID- 12556139 TI - Frequency of multiple inseminations in field-collected Anopheles gambiae females revealed by DNA analysis of transferred sperm. AB - We investigated the frequencies of single and multiple matings in field-collected female Anopheles gambiae by conducting microsatellite DNA analyses on the sperm contained within their spermatheca. Amplifcation by a polymerase chain reaction (PCR) at four loci allowed the detection of sperm extracts exhibiting more than two alleles per locus, thereby revealing the occurrence of multiple inseminations. Polyandry was found in six of 239 females examined, or 2.5% of the samples. Previous analyses of the molecular form of the sperm and female extracts using a PCR-based diagnostic procedure showed that two of these multiple inseminations involved cross-mating between two chromosomal/molecular forms of An. gambiae s.s. Thus polyandry occurred within-form in 1.7% of examined females while other multiple inseminations may be linked to processes of reproductive isolation between forms of An. gambiae. PMID- 12556140 TI - Short report: distribution and feeding preference of the sand flies Phlebotomus sergenti and P. papatasi in a cutaneous leishmaniasis focus in Sanliurfa, Turkey. AB - Sand flies (Diptera: Psychodidae) and rodents were collected in the endemic focus of urban cutaneous leishmaniasis in Sanliurfa, Turkey. Phlebotomus sergenti and P. papatasi represented 99% of the sand fly population. These flies were trapped in highest numbers in animal sheds, followed by cellars. However, P. sergenti was relatively more abundant in rooms. An enzyme-linked immunosorbent assay of the gut contents from blood-fed females detected immunoglobulins specific to birds and mammals, suggesting that both species are opportunistic feeders, although poultry is a frequent blood source of P. sergenti. Blood sources include black rats (Rattus rattus) and house mice (Mus domesticus); these rodents are abundant inside houses, and might have a role in parasite circulation. PMID- 12556141 TI - Risk mapping of visceral leishmaniasis: the role of local variation in rainfall and altitude on the presence and incidence of kala-azar in eastern Sudan. AB - Visceral leishmaniasis (VL) is a vector-borne disease highly influenced by environmental factors. A model was developed for mapping the distribution and incidence of VL in Gedaref State, eastern Sudan, in relation to different environmental factors. Geographical information systems (GIS) were used to extract and map regression results for environmental variables of 190 villages in Gedaref State, including rainfall, vegetation status, soil type, altitude, distance from river, topography, wetness indexes, and average rainfall estimates. VL incidence in each village was calculated from hospital records. By use of logistic and linear multivariate regression analyses, models were developed to determine which environmental factors explain variability in VL presence and incidence. We found that average rainfall and the altitude were the best predictors of VL incidence. The resulting models were mapped by GIS software predicting both VL presence or absence and incidence at any locality in Gedaref State. The results are discussed in relation to VL control. PMID- 12556142 TI - Duplex Doppler ultrasound of hepatic Schistosomiasis japonica: a study of 47 patients. AB - This study describes the ultrasound (US) appearances of the liver with hepatic schistosomiasis japonica (HSJ), and studies the portal hemodynamics in 47 patients with HSJ using duplex Doppler US over a period of 15 years. All patients but two were Chinese war veterans seen in Taiwan about 35-55 years after their presumed infection in Mainland China. The US presentations were reviewed. The data from Doppler portal flow studies were available for 39 patients with HSJ, and compared to data from Doppler portal flow studies in 40 normal healthy volunteers and to this data in 40 patients with postnecrotic cirrhosis. A typical "coarse reticular pattern" due to fibrosis in the whole liver was noted in 40 patients (85%). Other findings included periportal fibrosis (15%), septum-like fibrous bands extending to the liver capsule (32%), and an apparent nodular liver surface (19%). Splenomegaly was noted in seven patients. While coexisting hepatocellular carcinomas (HCC) were evident in three patients, and esophageal varices were found in three others, yet both conditions were found only in patients with positive hepatitis-B-surface antigen (HBsAg). Doppler flowmetry of the portal veins in HSJ patients showed a mean flow rate of 15.34 +/- 6.82 cm/sec, and a mean flow volume of 993.21 +/- 290.63 ml/min, both showed no significant difference from those in normal adults (p > 0.5). HSJ can be confidently diagnosed in patients with hepatic fibrosis when the hepatic pathology is presented as a coarse reticular pattern. The portal hemodynamics in HSJ patients who have been isolated from the infection site (for more than 35 years) are significantly different from portal hemodynamics in cirrhotic patients and are similar to those in healthy volunteers. PMID- 12556143 TI - Randomized, double-blind, placebo-controlled trial of oral artemether for the prevention of patent Schistosoma haematobium infections. AB - Artemether is an efficacious antimalarial drug that also displays antischistosomal properties. Laboratory studies have found that artemether curtails the development of adult worms of Schistosoma japonicum, S. mansoni and S. haematobium, and thus prevents morbidity. These findings have been confirmed in clinical trials for the former two parasites; administered orally once every 2 3 weeks, artemether significantly reduced the incidence and intensity of patent infections. Here, we present the first randomized, double-blind, placebo controlled trial of artemether against S. haematobium, done in a highly endemic area of Cote d'Ivoire. Urine specimens from 440 schoolchildren were examined over 4 consecutive days, followed by two systematic praziquantel treatments 4 weeks apart. S. haematobium-negative children were randomized to receive 6 mg/kg artemether (N = 161) or placebo (N = 161). Medication was administered orally for a total of six doses once every 4 weeks. Adverse events were assessed 72 hours after medication, and perceived illness episodes were monitored throughout the study period. Incidence and intensity of S. haematobium infections, and microhematuria and macrohematuria were assessed 3 weeks after the final dosing. We also monitored malaria parasitemia and treated positive cases with sulfadoxine pyrimethamine (SP). Oral artemether was well tolerated. The incidence of patent S. haematobium infections in artemether recipients was significantly lower than in placebo recipients (49% versus 65%, protective efficacy: 0.25, 95% CI: 0.08 0.38, P = 0.007). The geometric mean infection intensity in the artemether group was less than half that of the placebo recipients (3.4 versus 7.4 eggs/10 mL urine, P < 0.001). Heavy S. haematobium infections, microhematuria and macrohematuria, and the incidence of malaria parasitemia were all significantly lower in artemether recipients. In conclusion, previous findings of efficacy of artemether against S. japonicum and S. mansoni were confirmed for S. haematobium, although the protective efficacy was considerably lower. These findings enlarge the scope and potential of artemether and further contribute to discussions of its role as an additional tool for integrated schistosomiasis control. PMID- 12556144 TI - Long-term follow-up of treatment with diethylcarbamazine on anti-filarial IgG4: dosage, compliance, and differential patterns in adults and children. AB - We have followed a population in an area endemic for Brugia malayi for three years after intensive treatment with diethylcarbamazine (DEC). Microfilariae were cleared from the circulation within four months in all eligible study participants (n = 60). There appeared to be a strong correlation between the maximum reduction in specific IgG4 and the number of days drug was taken under supervision (p = 0.41, P < 0.001), indicating that high total dosage of DEC is necessary for optimal reduction of active infection. In individuals with good compliance (at least 180 mg/kg of body weight, n = 34), we observed variable IgG4 patterns. All pre-treatment IgG4+ children (9-14 years old) and 40% of the IgG4+ adult population (> or = 15 years old) showed a gradual decrease in anti-filarial IgG4; 53% of these showed complete clearance of worm burden by the end of the study. In contrast, another group of male IgG4+ adults showed IgG4 patterns that started to increase between nine months and two years after treatment, indicating either a partial efficacy of DEC that allowed recovery of resident adult worms or reinfection. PMID- 12556146 TI - Short report: Ancylostoma ceylanicum: exsheathment is not required for successful cryopreservation of third stage hookworm larvae. AB - Third-stage larvae (L3) of the human hookworm parasite Ancylostoma ceylanicum were cultured from the feces of infected hamsters and frozen for up to 100 days in liquid nitrogen. Upon thawing, viable larvae were recovered and used to inoculate naive hamsters. The larvae recovered from this second group of hamsters were used to inoculate a third group of naive animals, which demonstrated that the originally frozen larvae were successfully maintained for two full generations following thawing. These data suggest that exsheathment, which has previously been reported to be essential for successful cryopreservation, is not necessary for recovery of viable, infectious A. ceylanicum L3. PMID- 12556145 TI - Short report: Echinococcus granulosus from Xinjiang, PR China: cDNAS encoding the EG95 vaccine antigen are expressed in different life cycle stages and are conserved in the oncosphere. AB - The EG95-based vaccine protects sheep from infection with the dog tapeworm Echinococcus granulosus. The EG95 encoding gene is a member of a multigene family, several members of which are expressed in the oncosphere, believed to be the target of immunity induced by the vaccine. E. granulosus exhibits extensive intraspecific (strain) variation, and variability of the eg95 gene in different isolates of E. granulosus may directly impact the effectiveness of the EG95-based vaccine. We analyzed the eg95 gene from E. granulosus collected in Xinjiang, in northwest China, where hydatid disease is hyperendemic. The gene is expressed in oncospheres, protoscoleces, and immature and mature adult worms, and the eg95 gene family was shown to comprise two basic sequence types. Very limited sequence variation was evident in the EG95 protein from oncospheres. This high degree of sequence conservation predicts that the vaccine will continue to be effective in China and elsewhere. PMID- 12556147 TI - Case report: ophthalmomyiasis externa in Dallas County, Texas. AB - Ophthalmomyiasis externa is an uncommon condition in North America. If not recognized and managed accordingly, it can be complicated by the potentially fatal condition ophthalmomyiasis interna. Ophthalmomyiasis externa is mainly caused by the sheep bot fly Oestrus ovis; thus, it is more common in farming communities. We report a case of ophthalmomyiasis externa in a young woman from Dallas County, Texas, who had no known history of contact with farm animals. PMID- 12556148 TI - Increased apoptosis and expression of tumor necrosis factor-alpha caused by infection of cultured human monocytes with dengue virus. AB - Dengue (DEN) virus is responsible for one of the most significant viral diseases in tropical countries. Monocytes/macrophages (Mo/Mphi) are the major target cells for DEN virus. To determine the effects of the interaction between DEN virus and Mo/Mphi, human monocyte cultures were infected with DEN virus type 2. Apoptosis and production of tumor necrosis factor-alpha (TNF-alpha) and nitric oxide were measured in control and infected cultures. Virus was taken up by phagocytosis, but no membrane-coated pits at the virus attachment sites were observed. Increased number of apoptotic cells and increased production of TNF-a were observed in infected monocyte cultures. No increase in production of nitric oxide was observed. These results may be related to early primary viral infection, in which virus could induce apoptosis in monocytes, but monocytes may contribute to host defense mechanisms against virus by viral phagocytosis, phagocytosis of infected apoptotic cells, and the release of proinflammatory cytokines. PMID- 12556150 TI - Generation and characterization of cDNA clones from Sarcoptes scabiei var. hominis for an expressed sequence tag library: identification of homologues of house dust mite allergens. AB - Molecular studies on scabies, a disease of considerable human and veterinary significance, have been limited because of the difficulty of obtaining the causative organism Sarcoptes scabiei, the "itch mite." We have used skin from the bedding of crusted scabies patients as a source of mites for the construction of libraries of cDNAs from S. scabiei var. hominis in the bacteriophage lambda vector lambdaZAP express. Sequences of 145 clones established that the libraries predominantly contain sequences from S. scabiei, enabling a major sequencing program to begin. Among those sequenced to date, cDNAs encoding S. scabiei homologues of 3 house dust mite allergens-the M-177 apolipoprotein, glutathione S transferase, and paramyosin--were identified. The availability of cDNA libraries from S. scabiei var. hominis and S. scabiei var. vulpes and the emerging public sequence databases from both opens up new possibilities in scabies research. PMID- 12556151 TI - Genotyping of Balamuthia mandrillaris based on nuclear 18S and mitochondrial 16S rRNA genes. AB - Balamuthia mandrillaris is an opportunistically pathogenic ameba that causes fatal granulomatous amebic encephalitis (GAE) in vertebrates. Previous phylogenetic analyses that included the sequence of a single nuclear small subunit ribosomal RNA gene (18S or ssu rDNA) from this ameba suggested that Balamuthia is closely related to Acanthamoeba, another opportunistically pathogenic amebic genus, which includes multiple ssu rDNA genotypes. We tested whether this also is true for Balamuthia. The nuclear ssu rDNA from 4 isolates and the mitochondrial ssu rDNA from 7 isolates of B. mandrillaris have been sequenced. No variation in the nuclear rDNA sequences and low levels of variation in the mitochondrial rDNA were found. Both gene sequences were consistent with a single genotype for B. mandrillaris. The mitochondrial sequences of B. mandrillaris are unique and should be useful for development of genus-specific diagnostic probes for use with clinical, environmental, and archived specimens. PMID- 12556149 TI - Identification of a homologue of a house dust mite allergen in a cDNA library from Sarcoptes scabiei var hominis and evaluation of its vaccine potential in a rabbit/S. scabiei var. canis model. AB - Sarcoptes scabiei ("itch mite") causes scabies, a disease of considerable human and veterinary significance. Little work has been done at the molecular level because of the difficulty of obtaining mites. We have used mites in skin from the bedding of crusted scabies patients for the construction of a library of 10(5) cDNAs from S. scabiei var. hominis cloned in the vector pGEX4T-2. We describe the isolation by immunoscreening of 2 clones, one of which (Ssagl) is homologous to and cross-reactive with the house dust mite Euroglyphus maynei allergen M-177, an apolipoprotein from hemolymph. Immunohistochemistry revealed that it is located around the internal organs and cuticle of the mite and in eggs. Although it was not found to be protective in a challenge trial, the rabbits did not exhibit typical crust characteristics. This work shows that it is now possible to conduct such challenge trials with cloned scabies antigens. PMID- 12556152 TI - Enhancement of disease and pathology by synergy of Trichuris suis and Campylobacter jejuni in the colon of immunologically naive swine. AB - Campylobacter jejuni, a leading cause of bacterial gastroenteritis, has different age distribution and disease expression in developing and developed countries, which may be due to the endemnicity of infection and the age of acquisition of immunity. Differences in disease expression are not solely dependent on the C jejuni strain or virulence attributes. Another modulating factor in developing countries may be endemic nematode infections such as Trichuris, which drive type 2 cytokine responses and down-regulate type 1 immune responses. In this study, three-day-old germ-free pigs given dual infections with Trichuris suis and C jejuni had more frequent, more severe diarrhea and severe pathology than pigs given no pathogens, only T. suis, or only C jejuni. These pigs had significant hemorrhage and inflammatory cell infiltrates in the proximal colon where adult worms were found, and abscessed lymphoglandular complexes in the distal colon with intracellular C jejuni. Pigs given only C jejuni had mild clinical signs and pathology, and bacteria in feces or extracellular sites. Pigs given T. suis or no pathogens had no disease and minimal pathology. Thus, these agents synergized to produce significant disease and pathology, which was site specific. PMID- 12556153 TI - Leprosy affects facial nerves in a scattered distribution from the main trunk to all peripheral branches and neurolysis improves muscle function of the face. AB - Current literature rejects nerve release in leprous facial neuropathy and states that lesions are restricted to the peripheral zygomatic branches. Since there are approximately 500,000 patients with this disease throughout the world, we wanted to clarify the precise location of facial nerve's affection and the benefit of neurolysis. Our study showed that in patients with leprosy, the facial nerve's main trunk, the peripheral zygomatic branches, and all other branches were affected. Follow-up showed improvement in lagophthalmos and in misreinnervation, with no improvement in the control cohort. Nerve release improves muscle function in leprous facial neuropathy, provided surgery is performed on all affected segments. Intraoperative electroneurodiagnostics is an effective tool for detecting the most proximal site of lesion and ensuring effective surgery. PMID- 12556154 TI - Possible scrub typhus coinfections in Thai agricultural workers hospitalized with leptospirosis. AB - Possible coinfections with Orientia tsutsugamushi the causative agent of scrub typhus, were prospectively evaluated in rice farmers hospitalized with leptospirosis in Northeast Thailand. Of 22 adults with leptospirosis diagnosed by the microscopic agglutination test, 9 also had serologic evidence of scrub typhus. Of 9 individuals with possible coinfections, 5 had signs or symptoms typical of scrub typhus and atypical of leptospirosis. Patients who appeared to have mixed infections had significantly higher median platelet counts and significantly lower median serum bilirubin and creatinine concentrations (P < 0.05, Mann-Whitney U test) than did individuals with leptospirosis alone. One patient with serologic evidence of scrub typhus and leptospirosis was treated only with penicillin, to which scrub typhus is not sensitive. Respiratory distress worsened during therapy, and the patient died of respiratory failure. Physicians should consider the possibility of scrub typhus infection in leptospirosis patients who respond poorly to treatment or who have atypical disease manifestations. PMID- 12556155 TI - Subcellular localization of rickettsial invasion protein, InvA. AB - To understand further the molecular basis of rickettsial host cell invasion, Rickettsia prowazekii invasion gene homolog (invA) has been characterized. Our previous experiments have shown that InvA is an Ap5A pyrophosphatase, a member of the Nudix hydrolase family, which is up-regulated during the internalization, early growth phase, and exit steps during rickettsial mammalian cell infection. In addition to the molecular characterization, subcellular localization of InvA was investigated. InvA-specific antibodies were raised in mice and used for immunoelectron microscopy. The generated antibodies were shown to recognize InvA and by immunogold labeling showed InvA in the cytoplasm of rickettsiae. A cytoplasmic location for InvA would allow for a rapid response to any internal substance and efficient functioning in hydrolysis of toxic metabolic by-products that are accumulated in the rickettsial cytoplasm during host cell invasion. Protecting bacteria from a hazardous environment could enhance their viability and allow them to remain metabolically active, which is a necessary step for the rickettsial obligate intracellular lifestyle. PMID- 12556156 TI - Protective efficacy of the RTS,S/AS02 Plasmodium falciparum malaria vaccine is not strain specific. AB - RTS,S/AS02 is a recombinant protein malaria vaccine that contains a large portion of the C-terminal of the circumsporozoite protein (CSP) sequence of the NF54 isolate of Plasmodium falciparum fused to the hepatitis B virus surface antigen. It has been shown to induce significant protection to challenge infection with a homologous parasite strain in American volunteers. In a recently completed trial in semi-immune Gambian adults, vaccine efficacy against natural infection was 34% (95% confidence interval = 8-53%, P = 0.014) during the malaria season following vaccination. Breakthrough P. falciparum parasites sampled from vaccinated subjects and from controls were genotyped at two polymorphic regions of the csp gene encoding T cell epitopes (csp-th2r and csp-th3r) to determine if the vaccine conferred a strain-specific effect. The overall distribution of csp allelic variants was similar in infections occurring in vaccine and control groups. Also, the mean number of genotypes per infection in the RTS,S/AS02 group was not reduced compared with the controls. PMID- 12556157 TI - Serum cytokine profiles in patients with Plasmodium vivax malaria: a comparison between those who presented with and without hyperpyrexia. AB - Serum cytokine profiles in patients with Plasmodium vivax malaria who presented with and without hyperpyrexia were compared by a retrospective review of the medical records of the consecutive patients seen at the military hospitals near the demilitarized zone in the Republic of Korea from April 2000 through October 2001. Of 162 male patients studied, 120 (86.4%) presented with hyperpyrexia (i.e., an axillary temperature > or = 40 degrees C). The mean +/- SEM ages of the patients with and without hyperpyrexia were 21.5 +/- 0.14 and 21.9 +/- 0.39 years, respectively (P = 0.33). The mean +/- SEM concentrations of serum interleukin (IL)-6 (379.7 +/- 44.1 pg/mL versus 105.4 +/- 26.8 pg/mL; P = 0.002), IL-10 (583.4 +/- 58.2 pg/mL versus 142.4 +/- 39.7 pg/mL; P = 0.0001), and interferon-gamma (312.6 +/- 33.9 pg/mL versus 112.9 +/- 27.1 pg/mL; P = 0.0001) were significantly higher in patients with hyperpyrexia compared with those without hyperpyrexia. The mean +/- SEM concentrations of serum tumor necrosis factor-alpha were 155.5 +/- 54.5 pg/mL and 109.9 +/- 29.3 pg/mL (P = 0.27) in patients who presented with and without hyperpyrexia, respectively. Further studies are needed to examine whether serum concentrations of these cytokines also parallel their concentrations at the tissue sites of their production and action. PMID- 12556158 TI - Persistence of atovaquone in human sera following treatment: inhibition of Plasmodium falciparum development in vivo and in vitro. AB - Published pharmacokinetic data indicate that after treatment of patients with therapeutic doses of atovaquone/proguanil hydrochloride (Malarone, GlaxoSmithKline Research Triangle Park, NC), the plasma half-lives of these drugs are 70h and 15h, respectively. However, using two biologic assays (mosquito transmission and in vitro asexual stage development), we demonstrate here that sera from volunteers treated with atovaquone/proguanil retained activity against Plasmodium falciparum up to 6 weeks after such treatment. This activity was due to atovaquone, as administration of this drug alone replicated the data obtained with the combination. Most notably, asexual stage development of an atovaquone resistant strain (NGATV01) of P. falciparum was not inhibited by sera taken after atovaquone treatment. These data indicate that for atovaquone, biologic assays, though not quantitative, are more sensitive than the usual physicochemical assays. Also, persistence of atovaquone in plasma at low concentrations for long periods may increase the risk of resistant parasites arising. PMID- 12556159 TI - Placental monocyte infiltrates in response to Plasmodium falciparum malaria infection and their association with adverse pregnancy outcomes. AB - Maternal anemia and low birth weight (LBW) may complicate malaria in pregnancy, and placental monocyte infiltrates have been associated with LBW, and anecdotally with anemia. We examined placental pathology from 357 Malawian women. Intervillous monocyte infiltrates were frequent in placental malaria and were not seen in uninfected placentas. Histology was grouped according to a 5-point scale. Dense monocyte infiltrates and presence of intramonocytic malaria pigment were associated with anemia and LBW. Of factors associated with LBW and/or anemia in univariate analysis, gravidity (P = 0.002), number of antenatal clinic (ANC) visits (P < 0.001), malaria pigment in fibrin (P = 0.03), and monocyte malaria pigment (P = 0.0001) remained associated with lower birth weight by multivariate analysis. Associated with maternal anemia were HIV infection (P < 0.0001), intervillous monocyte numbers (P < 0.0001), number of ANC visits (P = 0.002), and recent febrile symptoms (P = 0.0001). Pigment-containing placental monocytes are associated with anemia and LBW due to malaria, and may have a causative role in their development. PMID- 12556160 TI - Mild and highly chemoselective oxidation of thioethers mediated by Sc(OTf)3. AB - [reaction: see text] Catalytic Sc(OTf)(3) greatly increases the efficiency of hydrogen peroxide mediated monooxidation of alkyl-aryl sulfides and methyl cysteine containing peptides. The method is high yielding, compatible with many widely used protecting groups, suitable for solid-phase applications and proceeds with minimum over-oxidation. PMID- 12556161 TI - Asymmetric synthesis of the putative structure of (-)-oryzoxymycin. AB - [reaction: see text] A short asymmetric synthesis of (2'S,5R,6R)-2'-[6-amino-5 hydroxy-1,3-cyclohexadiene-1-carbonyloxy]propionic acid, the enantiomer of the reported structure of (+)-oryzoxymycin is described. The reported spectral data does not match that obtained for synthetic "oryzoxymycin". PMID- 12556162 TI - Chemoselectively template-assembled glycoconjugates as mimics for multivalent presentation of carbohydrates. AB - [structure: see text] We report the highly efficient preparation of well-defined tetravalent glycoconjugates as potential tools for cell targeting and cell surface mimics. Our strategy is based on chemoselective assembly of aminooxy carbohydrates to a cyclic decapeptide template presenting aldehydes, namely, a "regioselectively addressable functionalized template" (RAFT). Preliminary recognition assays of tetramannosyl glycoconjugate (20) with specific lectin (Concanavalin A) using the fluorescence anisotropy method are also reported. PMID- 12556163 TI - Synthesis of oligonucleotides with a 2'-cap at the 3'-terminus via reversed phosphoramidites. AB - [reaction: see text] A method is presented for the synthesis of single compounds or small combinatorial libraries of oligonucleotides with 2'-acylamido-2' deoxyuridine residues at the 3'-terminus. Selection experiments identified the residue of anthraquinone-2-carboxylic acid as a "molecular cap" that increases the UV melting point of the duplex (5'-ACGCGU-3')(2) by up to 28 degrees C compared to the unmodified control duplex. PMID- 12556164 TI - Binding ability and self-assembly behavior of linear polymeric supramolecules formed by modified beta-cyclodextrin. AB - [structure: see text] The binding ability and self-assembly behavior of molecular interpenetration by newly synthesized mono[6-O-(4-formyl-phenyl)-beta cyclodextrin has been investigated, revealing the formation mechanism of modified cyclodextrin from solution aggregation to solid linear polymeric supramolecules. PMID- 12556166 TI - The tandem Heck-allylic substitution reaction: a novel route to lactams. AB - [reaction: see text] A novel route to substituted lactams has been developed using a tandem Heck-allylic substitution reaction. The palladium-catalyzed reaction between omega-olefinic N-tosyl amides and vinylic bromides affords in one step the substituted pyrrolidones and piperidones in 49-82% isolated yield. In addition, it is shown that an N-phenyl amide can act as a nucleophile in intramolecular allylic substitution reactions. PMID- 12556165 TI - Synthetic approaches to heavily lipidated phosphoglyceroinositides. AB - [reaction: see text] Naturally occurring phosphoinositide glycoconjugates are equipped with varied acyl residues that are important for their biological activity and biosynthesis. This paper reports that acylation at O2 of the myo inositol moiety can be achieved by stereocontrolled ortho ester rearrangement. Coupling to homo- or heterodiacylated glycerols was achieved via phosphoramidite methods, and exhaustive debenzylation by transfer hydrogenation afforded the deprotected phosphoglyceroinositides. The latter can be kept in chloroform solution at room temperature for over two months without migration of the inositol acyl group. PMID- 12556167 TI - Synthesis of highly substituted cyclopentenones via the [4 + 1] cycloaddition of nucleophilic carbenes and vinyl ketenes. AB - [reaction: see text] Vinyl ketenes have been shown to undergo [4 + 1] cycloaddition with a variety of nucleophilic carbenes to deliver highly substituted cyclopentenones as products. PMID- 12556168 TI - Total synthesis of (-)-pironetin. AB - [structure: see text] The asymmetric total synthesis of pironetin, a compound that shows plant growth regulatory activity, immunosuppressive as well as a remarkable antitumoral activity, is described. The approach involves the use of three very efficient Evans oxazolidinone-mediated syn-aldol condensations, a high yielding coupling between lithium acetylide ethylenediamine complex and a tosylate followed by methylation, and selective reduction to establish the C12 C13 (E) double bond. PMID- 12556169 TI - Total synthesis of (+)-benzastatin E via diastereoselective Grignard addition to 2-acylindoline. AB - [reaction: see text] A stereoselective total synthesis of (+)-benzastatin E (1) is described. The synthesis involves a diastereoselective Grignard addition to 2 acylindoline 2, which is derived from commercially available (S)-2 indolinecarboxylic acid (3). The unknown absolute configuration of (+)-1 is determined as (9S,10R). PMID- 12556170 TI - Synthesis of lariat-DNA via the chemical ligation of a dumbbell complex. AB - [reaction: see text] An efficient synthesis of a medium-sized DNA lariat through the chemical ligation of a Y-shaped dumbbell precursor is described. The methodology requires only commercially available phosphoramidites and reagents and affords regioisomerically pure lariat molecules. Characterization of the lariat by T(m) analysis reveals that the molecule displays markedly enhanced thermal stability and unimolecular association-dissociation kinetics, consistent with DNA dumbbell behavior. PMID- 12556171 TI - A concise, selective synthesis of the polyketide spacer domain of a potent bryostatin analogue. AB - [reaction: see text] A concise, asymmetric synthesis of the polyketide spacer domain portion (C1-C13) of a highly potent bryostatin analogue was developed. The route utilizes asymmetric hydrogenation methodology to install the C3, C5, and C11 stereocenters, while a substrate directed syn reduction sets the C9 stereocenter. The spacer domain 1 is obtained in 10 steps with a 25% overall yield and is readily incorporated into the synthesis of 2. PMID- 12556172 TI - The use of pH to influence regio- and chemoselectivity in the asymmetric aminohydroxylation of styrenes. AB - [reaction: see text] The pH-controlled Sharpless asymmetric aminohydroxylation (AA) of styrenes provides 1-aryl-2-amino ethanols (regioisomer B) with high enantio-, chemo-, and regioselectivity. As existing AA protocols typically give regioisomer A as the major reaction product when using carbamate nitrogen sources, this method is a convenient alternative for the selective production of regioisomer B. PMID- 12556173 TI - Iodine as a chemoselective reoxidant of TEMPO: application to the oxidation of alcohols to aldehydes and ketones. AB - [reaction: see text] Chemoselective alcohol oxidations using catalytic TEMPO and stoichiometric iodine as the terminal oxidant were studied. Iodine was compared to other positive halogens as the terminal oxidant and shown to be superior in cases of electron-rich and heteroaromatic rings. The new conditions were successfully applied to an important intermediate (2) in the synthesis of Losartan. PMID- 12556174 TI - Palladium-catalyzed reaction of aryl iodides with acetic anhydride. A carbon monoxide-free synthesis of acetophenones. AB - [reaction: see text] The palladium-catalyzed reaction of aryl iodides with acetic anhydride provides a straightforward and experimentally simple carbon monoxide free route to acetophenones. The reaction tolerates a wide range of functionalized aryl iodides. Acetophenones are isolated in excellent yield with a variety of neutral, slightly electron-rich, and slightly electron-poor aryl iodides, whereas moderate yields are obtained with aryl iodides containing strongly electron-withdrawing substituents. PMID- 12556175 TI - Asymmetric epoxidation catalyzed by N-aryl-substituted oxazolidinone-containing ketones: further evidence for electronic effects. AB - [reaction: see text] Ketones containing N-aryl-substituted oxazolidinones have been prepared and investigated for the epoxidation of cis-beta-methylstyrene, styrene, and 1-phenylcyclohexene. The attractive interaction between the phenyl group of the olefin and the oxazolidinone of the catalyst is enhanced by introducing an electron-withdrawing group onto the N-phenyl group of the catalyst. The information obtained gives a better understanding of the ketone catalyzed epoxidation. In addition, the easy preparation of some of the ketones makes them good candidates for practical use. PMID- 12556176 TI - Direct synthesis of ferrocenylmethylphosphines from ferrocenylmethyl alcohols and their application as ligands for room temperature Pd(0)-catalyzed Suzuki cross couplings of aryl bromides. AB - [reaction: see text] The direct, high-yield conversion of readily available ferrocenylmethyl alcohols to ferrocenylmethylphosphines and the application of ferrocenylmethylphosphines as efficient ligands for room temperature Suzuki cross coupling reaction of aryl bromides with phenylboronic acid are reported. The procedure of directly converting ferrocenylmethyl alcohols to ferrocenylmethylphosphines described here should find applications in the synthesis of many metallocenylmethylphosphines including optically active ones. PMID- 12556177 TI - Regioselective palladium-catalyzed arylation of 3-carboalkoxy furan and thiophene. AB - [reaction: see text] The regioselective palladium(0)-catalyzed arylation of 3 furoate and 3-thiophenecarboxylate esters with aryl bromides is described. Conditions were developed that allow for the selective synthesis of either 2-aryl or 5-aryl products. PMID- 12556178 TI - Probing the stereoselectivity of the Heck arylation of endocyclic enecarbamates with diazonium salts. Concise syntheses of (2S,5R)-phenylproline methyl ester and Schramm's C-azanucleoside. AB - [reaction: see text] The diastereoselectivity of the Heck arylation of several chiral, nonracemic, five-membered endocyclic enecarbamates with aryldiazonium tetrafluoroborates was evaluated. The cis selectivity observed for some enecarbamates bearing coordinating groups was explored in the concise synthesis of the (2S,5R)-(+)-phenylproline methyl ester, a scaffold for the nonpeptide cholecystokinin antagonist (+)-RP 66803, and in the synthesis of Schramm's potent antiprotozoan C-azanucleoside. PMID- 12556179 TI - Simultaneous and stereoselective formation of planar and axial chiralities in enantiopure sulfinyl iron diene complexes. AB - [reaction: see text] Enantiopure (1Z,3E)-1-sulfinyl dienes bearing an o dithianylphenyl group can be prepared and complexed with (bda)Fe(CO)(3) to afford the corresponding sulfinyl diene iron(0) tricarbonyl complexes. This diastereoselective complexation introduces planar and axial chirality simultaneously, with a high degree of facial selectivity as well as atropselectivity. Dynamic kinetic resolution is likely to be the origin of the atropselectivity. PMID- 12556180 TI - The second step of the halogen/metal exchange reaction. AB - [reaction: see text] The conversion of the iodine ate-complex 5 to the Grignard reagent 6 follows a rate law first order in 5 and in magnesium ions. In addition to this normal pathway of the halogen-metal exchange process, the conversion of 5 to 6 is catalyzed by a number of diverse "electrophiles". This points to a SET initiated radical chain process as a second pathway for the conversion of the iodine ate-complex 5 to the Grignard reagent 6. PMID- 12556181 TI - Iron-mediated electrochemical reaction of alpha-chloroesters with carbonyl compounds. AB - [reaction: see text] Reformatsky-type reactions have been performed efficiently using an electroassisted iron-complex catalysis. Valuable product such as beta hydroxyesters, ketones or nitriles are thus prepared with high yields. PMID- 12556182 TI - Diels-Alder reactions of 5-vinyl-1-acyl-2-aryl-2,3-dihydro-4-pyridones: regio- and stereoselective synthesis of octahydroquinolines. AB - [reaction: see text] Readily available 5-vinyldihydropyridones 2 undergo Diels Alder cyclization with various dienophiles to afford novel octahdroquinolines. The process is highly stereoselective and provides heterocyclic products containing synthetically useful functionality. PMID- 12556183 TI - A concise synthesis of the tricyclic skeleton of pleuromutilin and a new approach to cycloheptenes. AB - [reaction: see text] A short synthesis of the tricyclic skeleton of pleuromutilin is reported, featuring an unusually efficient 8-endo-trig radical cyclization of a xanthate precursor. In the course of this study, a one-carbon ring expansion leading to cycloheptenes was uncovered. PMID- 12556184 TI - Asymmetric halo aldol reaction (AHA). AB - [reaction: see text] The asymmetric halo aldol reaction (AHA) using Evans oxazolidinones as chiral auxiliaries has been established for tandem I-C/C-C bond formations. The new asymmetric reaction provides a practical approach to a variety of halo aldols of a non-Evans type that cannot be easily synthesized by other methods. Excellent diastereoselectivity (>95%) and yields (80-93%) have been obtained for eight examples. PMID- 12556185 TI - Novel rearrangement of a 2-aryl-3-alkyl-3H-indol-3-ol to a 1,4,5,6-tetrahydro-2,6 methano-1-benzazocin-3(2H)-one with implications for the biosynthesis of aspernomine. AB - [reaction: see text] Nominine (1) and aspernomine (2) are two biologically important indole diterpenoids that arise from a common digeranylindole precursor. The skeletal relationship of these two natural products was not heretofore understood. We have observed a novel rearrangement of 2-(2-bromophenyl)-3-(3 butenyl)-3H-indol-3-ol (5) to 7, which contains the uncommon 1,4,5,6-tetrahydro 2,6-methano-1-benzazocin-3(2H)-one ring system, under acidic conditions. This rearrangement suggests that aspernomine (2) may arise biosynthetically from nominine (1). PMID- 12556186 TI - Catalytic asymmetric synthesis of a 1-deoxy-1,1-difluoro-D-xylulose. AB - [reaction: see text] A new route, of potential strategic importance, to a difluorosugar analogue has been developed. Key steps included a Stille coupling and a highly regio- and enantioselective dihydroxylation of a highly substituted diene. Protecting groups were chosen to enhance the reactivity of the disubstituted allylic fragment in the AD reaction and allow deprotection under orthogonal conditions. PMID- 12556187 TI - Novel method for the immobilization of nucleotides. AB - [reaction: see text] A novel method for the immobilization of nucleotides has been developed. The strategy employs a highly reactive pyrrolidinium phosphoramidate zwitterion intermediate that undergoes nucleophilic attack by long-chain alkylamine-controlled pore glass (LCAA-CPG) to generate an immobilized nucleotide. Quantification of nucleotide loading was accomplished by acidic hydrolysis of the P-N bond and subsequent HPLC analysis of TMP in the presence of an internal standard. Typical nucleotide loadings of 51-59 micromol/g of support were observed. PMID- 12556188 TI - The first electroresponsive phenylazomethine macrocycles: highly preferential formation and regular molecular packing. AB - [structure: see text] Highly preferential formation of novel polyphenylazomethine macrocycles was achieved by further addition of TiCl(4) and/or the monomer during the course of the polycondensation. These macrocycles have unique structures based on the (E)/(Z)-conformation of the azomethine bonds, the extremely regular molecular-packing state, and the reversible redox properties by protic acid doping. PMID- 12556189 TI - Palladium-catalyzed cleavage of O/N-propargyl protecting groups in aqueous media under a copper-free condition. AB - [reaction: see text] A copper-free palladium-mediated cleavage of O/N-propargyl bonds in aqueous media has been investigated, affording a mild and convenient method for the deprotection of phenols and anilines. The methodology could be utilized for the selective removal of propargyl groups from aryl ethers and amines without affecting a variety of unprotected functional groups present in the substrates. The mechanism and scope of the reaction is discussed. PMID- 12556190 TI - Short and versatile route to a key intermediate for lactacystin synthesis. AB - [reaction: see text] A key intermediate 14 for the synthesis of lactacystin 1 has been constructed in four steps and 33% overall yield. The key steps involve cyclization of a suitably functionalized glutamic acid derivative and concomitant alkylation of the resulting beta,beta-diketoester system, C-acylation of the cyclic alpha-amidoketone 9, and decarboxylbenzylation of 12. Alkylation of a related beta,beta-diketoester 5 was additionally achieved with several electrophiles. PMID- 12556191 TI - Novel one-pot synthesis of 3-amino-1-alkenylphophonates by addition of imines to alkynylphosphonate titanium(II) complexes. AB - [reaction: see text] A new method of synthesis of 3-amino-1-alkenylphosphonates is described. It involves the addition of imines to the alkynylphosphonate titanium(II) complexes 2, which are prepared in situ from 1-alkynylphosphonates and Ti(O-i-Pr)(4)/2 equiv of i-PrMgCl. Compounds 4a-i were obtained regio- and stereoselectivily in high yields. PMID- 12556192 TI - Deoligomerization: a new route to lactams from unsaturated amides via radical oligomerization. AB - [reaction: see text] Triethylborane-initiated atom transfer radical oligomerization of N-allyl or N-(3-butenyl)iodoacetamides followed by treatment with hydrochloric acid and subsequent neutralization with K(2)CO(3) led to the formation of the corresponding 5-hydroxyl-substituted delta-lactams or caprolactams, respectively. This oligomerization-deoligomerization sequence serves as an alternative to the corresponding intramolecular cyclization reactions. PMID- 12556193 TI - Efficient and general synthetic method for preparing oligoenynes with either trans- or cis-olefinic configuration. AB - [reaction: see text] Efficient and practical synthesis of a variety of 1-iodo-4 trimethylsilylbut-1-en-3-yne derivatives with trans- and cis-olefinic configuration, and their repeated use as building blocks for the synthesis of trans- and cis-oligoenynes, respectively, are described. PMID- 12556194 TI - Synthesis and structure of a polyphenylene macrocycle related to "cubic graphite". AB - [structure: see text] Palladium-catalyzed coupling of 1,2-bis(4-bromophenyl) 3,4,5,6-tetraphenylbenzene and the corresponding hexaphenylbenzene bis(boronic acid) gave a mixture of linear and cyclic oligomers of hexaphenylbenzene. An X ray crystal structure of the tetrameric oligomer showed it to be the polyphenylene macrocycle 4 (C(168)H(112)). The roughly D(2) symmetric macrocycle contains a large central cavity, and it is one of the channel substructures of "phenylogous cubic graphite". PMID- 12556195 TI - Facile synthesis of 5,10-diaryl-5,10-dihydrophenazines and application to EL devices. AB - [structures: see text] An efficient method for the synthesis of 5,10 diaryldihydrophenazine was developed using a recently developed Pd(0)-mediated cross-coupling reaction. The products 1k and 3c showed excellent properties as hole injection materials in electroluminescent (EL) devices. PMID- 12556196 TI - Total synthesis of the formamicin aglycon, formamicinone. AB - [structure: see text] The total synthesis of formamicinone (2), the aglycone of formamicin (1), has been accomplished via the late-stage Suzuki cross-coupling of fragments 5 and 6, the macrolactonization of seco ester 14, and the Mukaiyama aldol reaction of aldehyde 3 and methyl ketone 4. An efficient and highly stereoselective second generation synthesis of vinyl iodide 6 is also described. PMID- 12556198 TI - Challenging but essential targets for genuine anti-ageing drugs. AB - Contrary to what one might conclude from the popular press, anti-ageing drugs do not yet exist, in the sense in which the term 'drug' is normally used. Since a drug is assumed to be effective against its target human pathology and since the vast majority of deaths in the developed world are from ageing-related causes, it is inappropriate to describe something as an anti-ageing drug unless one has good reason to believe that it will appreciably extend the life expectancy of those in the developed world who receive it. A drug that rejuvenates aspects of the aged body but does not increase life expectancy is an antifrailty drug, not an anti ageing one. This distinction is critical for decision makers in the drug discovery sphere because, while the market for antifrailty drugs (even unproven ones) is large, that for genuine anti-ageing drugs - which, as the author explains, are now foreseeable - will certainly be far larger. In this article, the author surveys the main aspects of age-related degeneration that are believed to be essential targets for genuine anti-ageing drugs - that is, without whose amelioration human life expectancy probably cannot be greatly increased - and some promising strategies for the design of such drugs. PMID- 12556199 TI - Cyclin-dependent protein kinases as therapeutic drug targets for antimalarial drug development. AB - Cyclin-dependent protein kinases (CDKs) have been attractive drug targets for the development of anticancer therapies due to their direct and crucial role in the regulation of cellular proliferation. Following this trend, CDKs have been pursued as potential drug targets for several other diseases. Structure-based drug design programmes have focused on the plasmodial CDKs to develop new candidate antimalarial compounds. This review discusses the most recent advances relating to three Plasmodium falciparum CDKs (PfPK5, PfPK6 and Pfmrk) as they are developed as antimalarial drug targets. CDKs are highly conserved, and focus must be placed upon the amino acid differences between human and plasmodial CDKs in order to develop specific inhibitors. Comparisons of the active sites of human and parasite CDKs reveal sequence and potential structural variations. Using sequence analysis, molecular modelling and in vitro drug screening, it is possible to identify and develop inhibitors that specifically target the plasmodial CDKs. These efforts are aimed at identifying new classes of CDK inhibitors that may be exploited for antimalarial drug development. PMID- 12556200 TI - Metalloproteinases: their role in arthritis and potential as therapeutic targets. AB - Irreversible degradation of articular cartilage is a major feature of the arthritides, and its prevention is a therapeutic goal which has been difficult to achieve. Enzymes from the matrix metalloproteinase and ADAMTS (a disintegrin, a metalloproteinase, and thrombospondin motif) families are key mediators of cartilage extracellular matrix destruction. Inhibition of metalloproteinase activity is therefore a conceptually attractive therapeutic strategy, although clinical efficacy has not yet been demonstrated. This review outlines the biology behind metalloproteinases as drug targets in the arthritides, and poses important questions for the future design of such therapies. PMID- 12556201 TI - Targeting monocyte chemoattractant protein-1 signalling in disease. AB - Monocyte chemoattractant protein-1 (MCP-1) has been implicated in many inflammatory and autoimmune diseases. The G-protein-coupled receptor CCR-2B is probably the most important MCP-1 receptor in vivo, and loss of MCP-1 effector function alone is sufficient to impair monocytic trafficking in inflammation models. MCP-1 signalling appears to be a relevant target, especially in rheumatoid arthritis (RA). In RA patients, MCP-1 is produced by synovial cells and infiltrating monocytes, plasma MCP-1 concentrations correlate with swollen joint count, and elevated serum MCP-1 concentrations were found in juvenile RA in patients with active disease. Modulation of MCP-1 signalling in experimental RA showed beneficial effects on inflammation and joint destruction. With respect to chronic neuroinflammation, a critical role for MCP-1 has been established in animal models for multiple sclerosis. In acute neuroinflammation, experimental evidence for a detrimental role of MCP-1 in stroke and excitotoxic injury has been found. Several selective small molecular weight CCR-2B antagonists and MCP-1 blocking antibodies have been described. The proof for the validity of targeting MCP-1 signalling in disease, however, has yet to be established in clinical trials. PMID- 12556202 TI - Emerging therapeutic targets in chronic heart failure: part II. AB - Chronic heart failure is characterised by functional deficiencies of the myocardium. Structural abnormalities of the left ventricular wall occur in many cases as a consequence of myocardial infarction (MI). The overburdened postMI heart is characterised by an active reorganisation of the remaining myocardium. Increased expression and activity of matrix metalloproteinases lead to altered composition and arrangement of the extracellular matrix, which is accompanied by eccentric hypertrophy of cardiomyocytes. The altered geometry of the heart muscle fosters biomechanical stress, driving the heart into a dead-end situation. Clearly, novel therapeutic concepts must be developed to reverse this process. Part II of the current review will focus on emerging therapeutic targets for small molecule therapeutics in the fields of cardiac remodelling and impaired survival of cardiomyocytes in the diseased heart. Finally, innovative therapeutic concepts for heart gene therapy and replacement options for destroyed post-MI myocardium using embryonic and adult stem cells are described. PMID- 12556203 TI - Apoptosis in the myocardium: much is still expected. AB - Apoptosis continues to be a controversial concept and subject of debate among scientists regarding its value as the basis for new therapeutic strategies. Today, it is widely accepted that the death of cardiac myocytes under a variety of conditions appears to be apoptotic based on a variety of criteria. However, the significance of these observations and how the insights into apoptotic molecular pathways may provide novel therapeutic targets remains to be determined. It is important to reconsider the pertinent underlying mechanisms of apoptosis regulation, and how these molecular pathways may be viewed in the functioning, intact heart. This knowledge can be applied in pursuit of practical goals in a search for new ways to prevent myocardial damage following such injuries as ischaemia/reperfusion or exposure to cardiotoxic drugs. Although recent literature contains reports of positive findings, there has not yet been a rigorous application of the model of apoptosis in the myocardium, and the potential for development of new therapeutic strategies is not yet understood. PMID- 12556204 TI - The not-so innocent bystander: the microenvironment as a therapeutic target in cancer. AB - The microenvironment in which cancer arises is often regarded as a bystander to the clonal expansion and acquisition of malignant characteristics of the tumour. However, a major function of the microenvironment is to suppress cancer, and its disruption is required for the establishment of cancer. In addition, tumour cells can further distort the microenvironment to promote growth, recruit non-malignant cells that provide physiological resources, and facilitate invasion. In this review, the authors discuss the contribution of the microenvironment, i.e., the stroma and its resident vasculature, inflammatory cells, growth factors and the extracellular matrix (ECM), in the development of cancer, and focus on two components as potential therapeutic targets in breast cancer. First, the ECM, which imparts crucial signalling via integrins and other receptors, is a first line barrier to invasion, modulates aggressive behaviour and may be manipulated to provide novel impediments to tumour growth. Second, the authors discuss the involvement of TGF-beta1 as an example of one of many growth factors that can regulate ECM composition and degradation and that play complex roles in cancer. Compared to the variable routes taken by cells to become cancers, the response of tissues to cancer is relatively consistent. Therefore, controlling and eliminating cancer may be more readily achieved indirectly via the tissue microenvironment. PMID- 12556205 TI - Analysing p53 tumour suppressor functions in mice. AB - Loss of tumour suppressor function is a common mechanistic step in deregulated cell growth and neoplasia. The p53 tumour suppressor gene is the most frequently mutated gene in cancer, and is inactivated in approximately 50% of human tumours. Mutation of p53 is also the predominant molecular basis of the Li-Fraumeni familial cancer susceptibility syndrome. p53 is a transcription factor that functions to regulate the integrity of the genome in response to DNA damage by inducing genes that promote cell cycle arrest, cell death, or repair of damaged DNA. These various effects exerted by p53 ensure that mutations do not pass on to subsequent generations, thus avoiding the presence of cells with multiple genetic hits that predispose the cell to neoplastic growth. Analysis of p53 functions using genetically-modified mice has complimented studies performed with human cancer tissue or cultured cells, and has greatly expanded knowledge about the role of p53 in tumour suppression. This finer understanding of p53 function has greatly facilitated research into small-molecule and other drug modifications of p53 activity as treatment modalities for the many human cancers bearing altered p53 function. This review will examine mouse models containing p53 modifications, and access the contribution of these studies to the understanding of p53-mediated tumour suppression. PMID- 12556207 TI - BAFF: B cell survival factor and emerging therapeutic target for autoimmune disorders. AB - The prevailing treatment strategies for autoimmune disorders employ global immunosuppressants that have harmful side effects with long-term use. A new vision for drug development relies on the generation of therapeutics that have specific and narrow targets, such as pathogenic cell populations. The cellular processes that initiate and maintain B cell dysregulation are not well understood and autoimmune disease results, in part, from the survival and activation of self reactive B cells. Such B cells produce tissue-damaging pathogenic autoantibodies. BAFF (B cell-activating factor belonging to the TNF family), a member of the TNF family of ligands, may play a role in B cell-mediated diseases. BAFF is a survival factor for peripheral B cells. When BAFF is overexpressed in mice, B cell number and immunoglobulin production is increased and an autoimmune-like phenotype is observed. Mouse models of lupus-nephritis have been shown to exhibit increased serum BAFF levels correlating with disease severity, and many autoimmune patients were found to have higher levels of circulating BAFF than healthy volunteers. Thus, modulating the level and activity of BAFF in these patients may alleviate symptoms associated with their disease. Several potential therapeutic inhibitors targeting BAFF are under investigation, including an anti BAFF antibody and receptor-Fc fusion proteins. PMID- 12556206 TI - CREB, memory enhancement and the treatment of memory disorders: promises, pitfalls and prospects. AB - The treatment of memory disorders, such as the gradual weakening of memory with age, the ravages of Alzheimer's disease and the cognitive deficits in various forms of mental retardation, may greatly benefit from a better understanding of the molecular and cellular mechanisms of memory formation. There is increasing interest in the possibility of pharmacologically enhancing learning and memory even in the absence of specific anatomically evident pathology. Substantial evidence in experimental systems ranging from molluscs to humans indicates that the cAMP response element binding protein (CREB) is a core component of the molecular switch that converts short- to long-term memory. Recent studies have greatly strengthened and refined our understanding of the role of CREB in learning and memory in mammals, in addition to providing greater insight into the molecular mechanisms of CREB regulation and function. This involvement of CREB and the upstream signalling pathways leading to its activation in learning associated plasticity makes them attractive targets for drugs aimed at improving memory function, in both diseased and healthy individuals. However, CREB and its close relatives cAMP response element modulator and activating transcription factor-1 are ubiquitous proteins with several critical functions. This creates hurdles that the authors believe may limit the usefulness of CREB per se as a target for the development of memory-enhancing drugs, and focus on components of the upstream signalling pathways or on specific downstream targets will be required. PMID- 12556208 TI - Highlights of frontiers in autoimmunity: fundamental aspects and clinical perspectives. AB - The aim of this meeting, on the shores of Balaton lake, was to discuss the ins, outs, ups and downs of autoimmune diseases. Participants were encouraged to discuss existing paradigms and to base their work on continuously reworked hypotheses derived from discoveries, rather than have it driven by theories based on a hypothesis. This event provided proof of the utility of modern approaches in defining the origin of this group of diseases as well as acting as an international forum for collaboration and dispute. Whilst highlighting the unique aspects of autoimmunity, this meeting suggested many avenues for future research and the design of novel strategies for immuno-intervention. PMID- 12556209 TI - Novel agents for the treatment of resistant Gram-positive infections. AB - Bacteria have proved themselves able to develop resistance to every antibiotic used clinically. Traditional agents used for treatment of serious infections caused by Gram-positive species have recently been supplemented with the introduction of linezolid, quinupristin-dalfopristin, several new quinolones and telithromycin. However, resistance to many of these agents has already been reported and, although each currently retains activity against the vast majority of clinical isolates of its target species, their long-term efficacy is uncertain. We must look to develop other compounds to replace and hopefully improve upon existing anti-Gram-positive agents. Daptomycin (a lipopeptide), oritavancin and dalbavancin (both second-generation glycopeptides) and ramoplanin (a glycolipodepsipeptide) are among the agents in advanced stages of development and, at present, many seem likely to proceed to licensing. In addition, it is encouraging that many agents active against novel bacterial targets have been discovered and are in earlier stages of development. In the next two decades, we should be optimistic that a regular flow of new anti-Gram-positive agents will enable us to offset the constant spectre of bacterial resistance. PMID- 12556210 TI - Recent developments in the treatment of sepsis. AB - Despite advances in supportive care, septic shock remains a major cause of morbidity and mortality. With the identification of the systemic inflammatory response as a major component in the pathogenesis of the septic shock syndrome, much of the recent work has focused on modulating this response. This includes antiendotoxin therapies in patients with Gram-negative sepsis, and therapies to modulate the pro-inflammatory mediators produced in response to infection, such as TNF-alpha, platelet-activating factor and complement. High-flow haemofiltration has the potential advantage of clearing both endotoxin and pro inflammatory mediators. Antithrombotic strategies have been investigated and have yielded the first major success in the treatment of sepsis with activated protein C. Nitric oxide produces the cardiovascular features of sepsis and investigators have looked at both reducing its production and mopping up the excess. Attempts to reduce apoptosis have been a new focus in the treatment of sepsis. There have also been recent developments in supportive care suggesting a role for vasopressin and replacement corticosteroid therapy. PMID- 12556211 TI - Recent developments in the discovery of hepatitis C virus serine protease inhibitors--towards a new class of antiviral agents? AB - Hepatitis C virus (HCV) infection is an epidemic disease and a significant worldwide health problem. Despite impressive improvements in the efficacy of the standard, interferon-based therapies, at present, the virus can not be eradicated in the majority of infected individuals. The last decade has witnessed a burst in our understanding of the molecular biology of HCV infection and lead to the identification of essential features of the viral genome that are being targeted for the development of specific antiviral agents. The non-structural protein 3 of the HCV genome harbours a serine protease domain that is essential for viral replication. This enzyme has been studied in great detail and the wealth of structural and functional data are presently nurturing drug development efforts. The peculiar active site structure of the enzyme imposes considerable obstacles to the development of small molecule inhibitors. However, the combination of creativity with the powerful tools of modern drug discovery has led to impressive progress in this field over the past few years and, as a result, the first compounds are now entering clinical trials. PMID- 12556212 TI - Novel agents and strategies to treat herpes simplex virus infections. AB - The quiet pandemic of herpes simplex virus (HSV) infection has plagued humanity since ancient times, causing mucocutaneous infection, such as herpes labialis and herpes genitalis. Disease symptoms often interfere with everyday activities and occasionally HSV infections are the cause of life-threatening or sight-impairing disease, especially in neonates and the immunocompromised patient population. After primary or initial infection the virus persists for life in a latent form in neurons of the host, periodically reactivating and often resulting in significant psychosocial distress for the patient. Currently, no cure is available. In the mid-1950s the first antiviral, idoxuridine, was developed for topical treatment of herpes disease and, in 1978, vidarabine was licensed for systemic use to treat HSV encephalitis. Acyclovir (Zovirax), a potent, specific and tolerable nucleosidic inhibitor of the herpes DNA polymerase, was a milestone in the development of antiviral drugs in the late 1970s. In the mid-1990s, when acyclovir became a generic drug, valacyclovir (Valtrex) and famciclovir (Famvir), prodrugs of the gold standard and penciclovir (Denavir), Vectavir), a close analogue, were launched. Though numerous approaches and strategies were tested and considerable effort was expended in the search of the next generation of an antiherpetic therapy, it proved difficult to outperform acyclovir. Notable in this regard was the award of a Nobel Prize in 1988 for the elucidation of mechanistic principles which resulted in the development of new drugs such as acyclovir. Vaccines, interleukins, interferons, therapeutic proteins, antibodies, immunomodulators and small-molecule drugs with specific or nonspecific modes of action lacked either efficacy or the required safety profile to replace the nucleosidic drugs acyclovir, valacyclovir, penciclovir and famciclovir as the first choice of treatment. Recently though, new inhibitors of the HSV helicase primase with potent in vitro antiherpes activity, novel mechanisms of action, low resistance rates and superior efficacy against HSV in animal models have been discovered. This review summarises the current therapeutic options, discusses the potential of preclinical or investigational drugs and provides an up-to-date interpretation of the challenge to establish novel treatments for herpes simplex disease. PMID- 12556213 TI - The therapeutic potential of CXCR4 antagonists in the treatment of HIV. AB - Since the identification of the chemokine receptors CXCR4 and CCR5 as co receptors for HIV-1 entry, several antagonists against these receptors have been synthesised. A highly selective CXCR4 antagonist, T22, and its downsized analogues T140 and TC14012, which inhibit X4-HIV-1 infection through their specific binding to CXCR4, have been identified. Besides T22 analogues, several other CXCR4 antagonists have been reported, such as AMD3100, ALX40-4C, KRH-1120 and AMD8664. Discovery of entry inhibitors, such as chemokine antagonists, may lead to the development of a new generation of antiHIV agents, since these inhibitors are thought to be useful for the clinical treatment of HIV-1-infected patients, especially at the late stage of treatment for AIDS patients developing multi-drug-resistant strains. In this review, recent research into CXCR4 antagonists in comparison with development of other antagonists is summarised. PMID- 12556214 TI - Delivery strategies for antiparasitics. AB - Optimisation of drug carrier systems and drug delivery strategies that take into account the peculiarities of individual infectious agents and diseases are key elements of modern drug development. In the following, different aspects of a rational design for antiparasitic drug formulation will be reviewed, covering delivery systems such as nano- and microparticles, liposomes, emulsions and microemulsions, cochleates and bioadhesive macromolecules. Functional properties for each carrier system will be discussed as well as their therapeutic efficacy for parasitic diseases, including leishmaniasis, human African trypanosomiasis, human cryptosporidiosis, malaria and schistosomiasis. Critical issues for the application of drug carrier systems will be discussed, focusing on biopharmaceutical and pathophysiological parameters such as routes of application, improvement of body distribution and targeting intracellularly persisting pathogens. PMID- 12556215 TI - Therapeutic potential of protease-activated receptor-1 antagonists. AB - The serine protease thrombin (EC 3.4.21.5) is central to the maintenance of haemostatic balance through its coagulant, anticoagulant and platelet activating properties. In addition, this enzyme affects numerous cellular responses in a wide variety of cells, such as cell proliferation, cytokine and growth factor release, lipid metabolism and tissue remodelling. A family of G-protein-coupled protease-activated receptors (PARs) mediates these cellular actions of thrombin. While thrombin can activate three of the four PAR family members, PAR-1 represents the primary thrombin-responsive receptor in human cells. The expression of PAR-1 in platelets, the vasculature and myocardium, in cells within atherosclerotic plaque and tissues after vascular injury, indicates that this receptor plays an important role during the response to tissue injury and associated inflammatory processes. With the development of PAR-deficient mice and small-molecule antagonists, it is now clear that intervening in processes mediated by PAR-1 presents a new approach to treating a variety of disorders dependent on thrombin generation, including thrombosis and restenosis. The full potential of PAR-1 antagonists has yet to be realised, but the promise of novel therapeutics that modulate receptor function rather than thrombin's proteolytic activity, provides an alternative and, perhaps, more desirable means to dampen the pathological effects of thrombin. PMID- 12556216 TI - PTP1B inhibitors as potential therapeutics in the treatment of type 2 diabetes and obesity. AB - Coordinated tyrosine phosphorylation is essential for signalling pathways regulated by insulin and leptin. Type 2 diabetes and obesity are characterised by resistance to hormones insulin and leptin, possibly due to attenuated or diminished signalling from the receptors. Pharmacological agents capable of inhibiting the negative regulator(s) of the signalling pathways are expected to potentiate the action of insulin and leptin and therefore be beneficial for the treatment of Type 2 diabetes and obesity. A large body of data from cellular, biochemical, mouse and human genetic and chemical inhibitor studies have identified protein tyrosine phosphatase 1B (PTP1B) as a major negative regulator of both insulin and leptin signalling. In addition, evidence suggests that insulin and leptin action can be enhanced by the inhibition of PTP1B. Consequently, PTP1B has emerged as an attractive novel target for the treatment of both Type 2 diabetes and obesity. The link between PTP1B and diabetes and obesity has led to an avalanche of research dedicated to finding inhibitors of this phosphatase. With the combined use of structure and medicinal chemistry, several groups have demonstrated that it is feasible to obtain small-molecule PTP1B inhibitors with the requisite potency and selectivity. The challenge for the future will be to transform potent and selective small molecule PTP1B inhibitors into orally available drugs with desirable physicochemical properties and in vivo efficacies. PMID- 12556217 TI - Friedreich's ataxia: iron chelators that target the mitochondrion as a therapeutic strategy? AB - Friedreich's ataxia (FA) is a severe inherited spinocerebellar ataxia that primarily affects the nervous system and heart leading to early confinement in a wheelchair and death. The gene defective in FA, FRDA, encodes a mitochondrial protein known as frataxin. A triplet repeat expansion within intron 1 of the FRDA gene results in a marked decrease in frataxin expression. Over the last 5 years it has become clear that this results in mitochondrial iron accumulation that generates oxidative stress and results in damage to critical biological molecules. Drugs that reduce oxidative stress have a limited effect on the progression and pathology of the disease, probably because these agents cannot remove the iron accumulation. In this review, the potential of iron chelators, namely the 2-pyridylcarboxaldehyde isonicotinoyl hydrazone (PCIH) analogues, as agents to remove mitochondrial iron deposits is discussed. These ligands have been specifically designed to enter and target mitochondrial iron pools, which is a property lacking in desferrioxamine, the only chelator in widespread clinical use. This latter drug may not have any beneficial effect in FA patients, probably because of its hydrophilicity that prevents mitochondrial access. Indeed, standard chelation regimens will probably not work in FA, as these patients do not exhibit gross iron-loading. Considering that there is no effective treatment for FA, it is essential that the therapeutic potential of iron chelators that target mitochondrial iron pools is assessed experimentally. PMID- 12556218 TI - Sjogren's syndrome: evolving therapies. AB - Sjogren's syndrome (keratoconjunctivis sicca) is a relatively common disorder with incidence of approximately 0.5% of adult women. It has both local (ocular and oral) features as well as systemic manifestations. There has been recent FDA approval of agents to stimulate salivation (pilocarpine and cevimeline) and studies are in progress to determine their role in the treatment of dry eye. New therapies are in clinical trials for ocular manifestations with the most interest focused on topical cyclosporin A and purinogenic receptor agonists. In oral therapy, topical human interferon has reported encouraging results in short-term studies. However, the high placebo response (probably reflecting the beneficial response of mechanical stimulation of the buccal mucosa by the lozenge) and the response to much cheaper therapies (such as acid maltose lozenges) may offer safer and cheaper alternatives. For systemic disease, there is interest in tumour necrosis factor inhibitors. However, the cost-effectiveness and safety of biological agents needs longer term follow up, as they appear much less dramatic in their effect on systemic lupus erythematosus or Sjogren's syndrome than in rheumatoid arthritis. PMID- 12556219 TI - Development potential of rifalazil. AB - Rifalazil represents a new generation of ansamycins that contain a unique four ring structure. Originally rifalazil was developed as a therapeutic agent to replace rifampin as part of a multiple drug regimen in the treatment of tuberculosis. As a result of its superior antimicrobial activity and high intracellular levels, rifalazil has potential to treat indications caused by the intracellular pathogen, Chlamydia trachomatis, which causes non-gonococcal urethritis and cervicitis, often leading to pelvic inflammatory disease. Rifalazil also has potential to treat the related microorganism, Chlamydia pneumoniae, which may be involved in chronic inflammatory processes thought to be partly responsible for atherosclerosis. Due to its favourable antimicrobial spectrum and other positive attributes, rifalazil may also prove valuable in the treatment of gastric ulcer disease, caused by Helicobacter pylori, and antibiotic associated colitis, the result of toxin production following the growth of Clostridium difficile in the colon. The potential value of rifalazil in the treatment of these indications will be assessed in human clinical trials. PMID- 12556220 TI - Substrate reduction therapy: miglustat as a remedy for symptomatic patients with Gaucher disease type 1. AB - Gaucher disease (GD) is an inborn error of glycosphingolipid metabolism, associated with multisystemic manifestations resulting from the lysosomal accumulation of an incompletely degraded material (glucosylceramide) within cells of monocyte/macrophage lineage. In the majority of GD patients with non neurological involvement (type 1), the clinical features are dominated by haematological, visceral (hepatic and splenic) and skeletal complications. Infrequently, cardiopulmonary involvement may develop and lead to major morbidity. Investigations of substrate turnover within cellular compartments, such as the lysosome, indicate that progressive tissue storage occurs when the activity of the responsible hydrolytic enzyme declines. Thus, the provision of functional enzyme to deficient cells is a straightforward means of achieving metabolic correction. Conversely, the concentration of stored substrate within cells bearing particular affected enzymes (i.e., expressing residual activity) may be controlled by a reduction of the 'load'. This means that metabolic homeostasis can also be restored by restricting the amount of substrate presented to disease cells to a level that can be hydrolysed by the existing enzyme activity. Proof-of-concept for both approaches has been demonstrated through enzyme replacement therapy using alglucerase/imiglucerase and more recently, substrate reduction therapy by miglustat. Enzyme replacement therapy is proven to be safe and effective in the treatment of GD type 1, establishing imiglucerase as the current standard of care. The experience with substrate reduction therapy, specifically miglustat, is limited but encouraging. This review is an attempt to examine the potential role of this latter approach in light of current patient management. The consideration of miglustat as a therapeutic option requires the appropriate selection of patients (amongst those unwilling or unsuitable to receive enzyme replacement therapy), a definition of the therapeutic objectives and monitoring not only for response but potential adverse effects. PMID- 12556221 TI - Indisulam: an anticancer sulfonamide in clinical development. AB - Indisulam (N-(3-chloro-7-indolyl)-1,4-benzenedisulfonamide, E7070) is a novel sulfonamide anticancer agent in clinical development for the treatment of solid tumours. Its mechanism of action is multifactorial, as indisulam arrests cell cycle in the G1 phase, strongly inhibits carbonic anhydrase, a critical enzyme involved in many physiological processes and whose association with cancer became obvious in the last period, and markedly alters gene expression levels of at least 60 transcripts. Four Phase I clinical trials gave promising results, showing the compound to possess nonlinear pharmacokinetics. Presently this compound is in Phase II trials in Europe and USA for the treatment of solid tumours. PMID- 12556222 TI - Dipyrimidine-based inhibitors of HIV-1 integrase. PMID- 12556223 TI - International congress on hormonal steroids and hormones in cancer. PMID- 12556224 TI - Detection of 3-hydroxykynurenine in a plant pathogenic fungus. AB - A redox-active compound has been purified from the barley powdery mildew fungus Blumeria ( Erysiphe ) graminis f. sp. hordei. A combination of spectrophotometry, MS and NMR has identified it as 3-hydroxykynurenine (3OHKyn). This compound, never previously detected in any fungus or pathogen, is best known for its role in vertebrate cataracts. It is found abundantly in developing and germinating spores and also in runner hyphae. Two roles for 3OHKyn are discussed: first, the presence of active oxygen species would enable 3OHKyn to cross-link the spore chemically with the plant. Secondly, it may be acting as an UV protectant and an antioxidant. PMID- 12556225 TI - Tailoring tissue inhibitor of metalloproteinases-3 to overcome the weakening effects of the cysteine-rich domains of tumour necrosis factor-alpha converting enzyme. AB - Tumour necrosis factor-alpha (TNF-alpha) converting enzyme (TACE) is a membrane anchored, multiple-domain zinc metalloproteinase responsible for the release of the potent pro-inflammatory cytokine, TNF-alpha. The extracellular part of the active enzyme is composed of a catalytic domain and several cysteine-rich domains. Previously, we reported that these cysteine-rich domains significantly weakened the inhibitory potency of the N-terminal-domain form of tissue inhibitor of metalloproteinases-3 (N-TIMP-3). In the present paper, we describe a novel strategy developed to overcome this weakening effect. We have engineered a new generation of N-TIMP-3 mutants that are capable of withstanding the repulsion of the cysteine-rich domains by the formation of electrostatic bonds with the catalytic domain of the enzyme. These N-TIMP-3 mutants displayed markedly improved binding affinity with the soluble extracellular domain form of recombinant TACE. With K (i) (app) values of <0.1 nM, these mutants were dramatically better than the wild-type N-TIMP-3 [K (i) (app) 1.7 nM]. We accounted for this by proposing that Glu(31), an acidic residue situated at the base of the AB-loop of N-TIMP-3, is drawn into contact with Lys(315), a prominent basic residue adjacent to the TACE catalytic site. The mutagenesis strategy involved reorientation of the edge of N-TIMP-3; in particular, the beta-strand A where Glu(31) was located. Further expression of one of the mutants, Lys(26/27/30/76)-->Glu, in a mammalian expression system confirmed that TIMP-3 associates with the extracellular matrix via its C-terminal domain. PMID- 12556226 TI - Thioredoxin peroxidases can foster cytoprotection or cell death in response to different stressors: over- and under-expression of thioredoxin peroxidase in Drosophila cells. AB - Recently, we identified a set of five genes constituting the peroxiredoxin gene family in Drosophila melanogaster [Radyuk, Klichko, Spinola, Sohal and Orr (2001) Free Radical Biol. Med. 31, 1090-1100]. This set includes two abundant thioredoxin peroxidase (TPx) species, namely Drosophila peroxiredoxin DPx-4783, a cytosolic TPx and DPx-5037, a mitochondrial TPx. Overexpression of either one of them in Drosophila S2 cells conferred increased resistance to toxicity induced by hydrogen peroxide, paraquat or cadmium. To understand further the functional roles of these enzymes in vivo, we report in the present study the effects of decreased expression, using RNA interference, on the response of S2 cells to different stressors. When either of the TPxs was blocked, cells became relatively more susceptible to oxidative stress caused by exposure to hydrogen peroxide or paraquat, but were unaffected when challenged with copper and heat stress. In contrast, TPx overexpressing cells were more susceptible to copper and heat stress when compared with control cells and exhibited DNA fragmentation. Furthermore, when cells were supplemented with N -acetyl-L-cysteine together with copper, there was a clear negative effect on cell survival, which was exacerbated by TPx overexpression. Manipulations in the levels of TPxs demonstrated that, under different stress conditions, these enzymes might have both beneficial and detrimental effects on Drosophila cell viability. PMID- 12556229 TI - To err is human. PMID- 12556227 TI - The internal alternative NADH dehydrogenase of Neurospora crassa mitochondria. AB - An open reading frame homologous with genes of non-proton-pumping NADH dehydrogenases was identified in the genome of Neurospora crassa. The 57 kDa NADH:ubiquinone oxidoreductase acts as internal (alternative) respiratory NADH dehydrogenase (NDI1) in the fungal mitochondria. The precursor polypeptide includes a pre-sequence of 31 amino acids, and the mature enzyme comprises one FAD molecule as a prosthetic group. It catalyses specifically the oxidation of NADH. Western blot analysis of fungal mitochondria fractionated with digitonin indicated that the protein is located at the inner face of the inner membrane of the organelle (internal enzyme). The corresponding gene was inactivated by the generation of repeat-induced point mutations. The respiratory activity of mitochondria from the resulting null-mutant ndi1 is almost fully inhibited by rotenone, an inhibitor of the proton-pumping complex I, when matrix-generated NADH is used as substrate. Although no effects of the NDI1 defect on vegetative growth and sexual differentiation were observed, the germination of both sexual and asexual ndi1 mutant spores is significantly delayed. Crosses between the ndi1 mutant strain and complex I-deficient mutants yielded no viable double mutants. Our data indicate: (i) that NDI1 represents the sole internal alternative NADH dehydrogenase of Neurospora mitochondria; (ii) that NDI1 and complex I are functionally complementary to each other; and (iii) that NDI1 is specially needed during spore germination. PMID- 12556230 TI - Genetic heterogeneity of Icelanders. AB - Recently statements have been made about a special 'genetic homogeneity' of the Icelanders that are at variance with earlier work on blood groups and allozymes. To validate these claims an extensive reanalysis was undertaken of mtDNA variation by examining primary data from original sources on 26 European populations. The results show that Icelanders are among the most genetically heterogeneous Europeans by the mean number of nucleotide differences as well as by estimates of theta parameters of the neutral theory. The distribution of pairwise differences in general has the same shape as European populations and shows no evidence of bottlenecks of numbers in Iceland. The allelic frequency distribution of Iceland is relatively even with a large number of haplotypes at polymorphic frequencies contrasting with other countries. This is a signature of admixture during the founding or history of Iceland. Assumptions of models used to simulate number of haplotypes at sampling saturation for comparing populations are violated to different degrees by various countries. Anomalies identified in data in previous reports on Icelandic mtDNA variation appear to be due to errors in publicly accessible databases. This study demonstrates the importance of basing analyses on primary data so that errors are not propagated. Claims about special genetic homogeneity of Icelanders are not supported by evidence. PMID- 12556231 TI - Genetic polymorphisms in the Renin-Angiotensin system in high-altitude and low altitude Native American populations. AB - Cardiovascular disease (CVD) is reportedly less common in high-altitude native populations than in lowlanders. To some extent, this is due to cultural and demographic factors; however, increased cardiovascular efficiency contributing to hypoxia adaptation may also be involved. Numerous genetic variants have been associated with cardiovascular health. If the decreased incidence of CVD in modern high-altitude populations reflects selective pressures having favoured the transmission of these alleles in their antecedents, it would be expected that these alleles would be more common in highlanders than in lowlanders. We tested this hypothesis by determining the allele frequencies of five polymorphic loci in genes encoding components of the renin-angiotensin system (RAS) that have alleles associated with hypertension and cardiovascular disease in a high-altitude native Andean population, Quechua from the Peruvian altiplano, and in a lowland Amerindian population, Maya from the Yucatan peninsula. The polymorphisms examined were 1) the insertion/deletion polymorphism in intron 16 of the angiotensin converting enzyme (ACE) gene; 2) the A/G2350 transition (ACE-8) in intron 17 of the ACE gene; 3) the A/C1166 transversion in the 3' untranslated region of the angiotensin II receptor (type 1) gene (AGTR1); 4) the G/AI9-83 transition in intron 8 of the renin gene (REN); and 5) the T/C704 (Met235Thr) transition mutation in angiotensinogen (AGT). There was no evidence for an over representation of the RAS alleles associated with cardiovascular fitness in the high-altitude Amerindian population when compared to the lowland Amerindian population. PMID- 12556232 TI - Haplotype structure of TP53 locus in Indian population and possible association with head and neck cancer. AB - It has been proposed that a constellation of three TP53 polymorphisms (intron 3 16 bp duplication, codon 72 BstUI, and intron 6 Nci I RFLP at nt 13494) constitute a haplotype predictive of increased cancer risk. We have estimated the allele frequency of these polymorphisms in three endogamous Indian ethnic populations from three different geographic locations (viz. Iyer from south India, Brahmin from central India and Mahishya from eastern India), as well as in head and neck squamous cell carcinoma (HNSCC) patients, and in ethnically matched normal individuals from the eastern region of India. The genotype distributions and allele frequencies of the three polymorphisms in all but one population, as well as in patients, showed a good fit to Hardy-Weinberg equilibrium. Strong linkage disequilibria were observed between all loci in every population examined, except for the 16bp-Nci I haplotype in the Mahishya population. The Mahishya population differed significantly from the other two populations with respect to differences in allele frequency and haplotype frequency. Although there were no significant differences in genotypic frequency at any of the loci between HNSCC patients and the matched control population, the minor allele frequency of codon 72 and intron 3 16 bp polymorphisms showed significant variation. Variation in overall haplotype frequency between patients and normal individuals was significant (p = 0.036) when two rare haplotypes 2-1-2 and 1-2-1 were combined. The rare haplotype 2-1-2 was found to be modestly over represented in HNSCC patients as compared to normal individuals. PMID- 12556233 TI - CARD15/NOD2 risk alleles in the development of Crohn's disease in the Australian population. AB - We have previously reported strong evidence for linkage between IBD1 and Crohn's disease (CD) in Australian Crohn's disease families. Three risk alleles for Crohn's disease, (Arg702Trp (C/T), Gly908Arg (G/C) and 980fs981 (-/C), were recently identified in the CARD15/NOD2 gene on chromosome 16, implicating this as the IBD1 locus. Using a novel diagnostic PCR-RFLP, we have examined the frequency of these alleles in 205 multiplex IBD families, 107 sporadic Crohn's disease cases and 409 normal individuals. We demonstrate that the three risk alleles are more frequent in Crohn's disease, than in controls, with allelic frequencies of 0.11, 0.02 and 0.07 respectively. Heterozygosity for individual variants conferred a three-fold increase in risk for Crohn's disease while substantially higher risks were associated with being homozygous or compound heterozygous. Despite a significantly lower population allele frequency for the frameshift mutation than reported by other groups, we see a similar contribution by this allele to the risk of developing Crohn's disease. While the three risk alleles influence susceptibility to Crohn's disease in Australia, we show that these alleles do not fully explain the linkage evidence and suggest that there are very likely additional IBD1 susceptibility alleles yet to be described in Australian CD at the NOD2 locus. We also show a second linkage peak in Australian CD that provides some support for a second disease susceptibility locus on chromosome 16. PMID- 12556234 TI - Autosomal microsatellite and mtDNA genetic analysis in Sicily (Italy). AB - DNA samples from 465 blood donors living in 7 towns of Sicily, the largest island of Italy, have been collected according to well defined criteria, and their genetic heterogeneity tested on the basis of 9 autosomal microsatellite and mitochondrial DNA polymorphisms for a total of 85 microsatellite allele and 10 mtDNA haplogroup frequencies. A preliminary account of the results shows that: a) the samples are genetically heterogeneous; b) the first principal coordinates of the samples are correlated more with their longitude than with their latitude, and this result is even more remarkable when one outlier sample (Butera) is not considered; c) distances among samples calculated from allele and haplogroup frequencies and from the isonymy matrix are weakly correlated (r = 0.43, P = 0.06) but such correlation disappears (r = 0.16) if the mtDNA haplogroups alone are taken into account; d) mtDNA haplogroups and microsatellite distances suggest settlements of people occurred at different times: divergence times inferred from microsatellite data seem to describe a genetic composition of the town of Sciacca mainly derived from settlements after the Roman conquest of Sicily (First Punic war, 246 BC), while all other divergence times take root from the second to the first millennium BC, and therefore seem to backdate to the pre-Hellenistic period. A more reliable association of these diachronic genetic strata to different historical populations (e.g. Sicani, Elymi, Siculi), if possible, must be postponed to the analysis of more samples and hopefully more informative uniparental DNA markers such as the recently available DHPLC-SNP polymorphisms of the Y chromosome. PMID- 12556235 TI - The study of APOA1, APOC3 and APOA4 variability in healthy ageing people reveals another paradox in the oldest old subjects. AB - The genes coding for apolipoprotein A1 (APOA1), apolipoprotein C3 (APOC3) and apolipoprotein A4 (APOA4) are tandemly organised within a short region on chromosome 11q23-q24. Polymorphisms of these genes have been extensively investigated in lipoprotein disorders and cardiovascular diseases, but poorly investigated in healthy ageing. The aim of this study was to describe possible modifications of the APOA1, APOC3, and APOA4 gene pool by cross-sectional studies carried out in a healthy ageing population whose ages ranged from 18 to 109 years (800 subjects, 327 males and 473 females, free of clinically manifested disease, and with emato-chemical parameters in the norm). APOA1-MspI-RFLP (-75 nt from the transcription starting site), APOC3-SstI-RFLP (3'UTR, 3238 nt), and APOA4-HincII RFLP (Asp127/Ser127) were analysed according to age and sex. A significant age related variation of the APOA1 gene pool was observed in males. An analysis of the allele average effect exerted by APOA1-MspI-RFLP A/P alleles (Absence/Presence of the restriction site) on lipidemic parameters in 46-80 year old males showed that allele A decreased, while allele P significantly increased, serum LDL-cholesterol. Unexpectedly, the P allele was over-represented in the group of the oldest old subjects, thus giving evidence of another "genetic paradox of centenarians". PMID- 12556236 TI - Seven novel acid sphingomyelinase gene mutations in Niemann-Pick type A and B patients. AB - We have analyzed acid sphingomyelinase (SMPD1; E.C. 3.1.4.12) gene mutations in four Niemann-Pick disease (NPD) type A and B patients of Turkish ancestry and in three patients of Dutch origin. Among four NPD type A patients we found two homozygotes for the g.1421C > T (H319Y) and g.3714T > C (Y537H) mutations and two compound heterozygotes, one for the g.3337T > C (F463S) and g.3373C > T (P475L) mutations and the other for the g.84delC (G29fsX74) and g.1208A > C (S248R) mutations. One of the type B patients was homozygous for the g.2629C>T (P371S) mutation. The last two type B patients were homozygotes for the common g.3927_3929delCGC (R608del) mutation. The G29fsX74, S248R, H319Y, P371S, F463S, P475L and Y537H SMPD1 mutations are all novel and were verified by PCR/RFLP and/or ARMS. All of the identified mutations are likely to be rare or private, with the exception of R608del which is prevalent among NPD type B patients from the North-African Maghreb region. Geographical and/or social isolation of the affected families are likely contributing factors for the high number of homozygotes in our group. PMID- 12556237 TI - Gene admixture in the Costa Rican population. AB - The general population of Costa Rica has sometimes been considered to be the product of an amalgamation of groups of diverse origin. To determine the magnitude of accumulated admixture since Spanish colonization, 11 classic genetic markers were analyzed in a total of 2196 individuals originating from five distinct regions of the country. A maximum likelihood approach was used. The proportions of genes of European, Amerindian and African ancestry were found to be 61%, 30% and 9% of the total population, respectively. Variation was observed at a regional level, with an increased European influence in the North (66%) and Central (65%) regions. Meanwhile an increase in Amerindian ancestry was found in the South (38%), and a higher incidence in the contribution of African genes was detected in the coastal regions (13% in the Atlantic and 14% in the North Pacific). A principal component (PC) analysis showed that 76% of the existing variability can be explained by the first two PCs, which is in agreement with the variations observed in the admixture process by geographic area. It has been concluded that the Costa Rican population is truly trihybrid, similar to populations in other Latin American countries; however, it differs from them fundamentally by the proportion of gene flow from ancestral populations. PMID- 12556238 TI - Simultaneous detection of ACP1 and GC genotypes using PCR/SSCP. AB - The classical enzyme and protein markers ACP1 and GC have gained new importance because of the biological functions of their gene products. ACP1 encodes a low molecular weight enzyme which is now recognized as a phosphotyrosine phosphatase with a role in the regulation of signal transduction pathways, and GC-globulin acts both as a transporter of vitamin D and as a plasma actin scavenger and plays a role in macrophage activation. These two polymorphisms were phenotyped for decades on the basis of electrophoretic isozyme or protein patterns; the gene structures are now known. Nucleotide substitutions determining the common alleles are close enough at each locus to be contained in one short PCR product. We have developed a simple, rapid and reliable multiplex method based on PCR and SSCP which allows the simultaneous determination of the common ACP1 and GC genotypes. PMID- 12556239 TI - Tuberous sclerosis: from tubers to mTOR. AB - Tuberous sclerosis (TSC) is an autosomal dominant hamartoma syndrome whose causative genes (TSC1 and TSC2) were identified 5 and 9 years ago respectively. Their encoded proteins are large, and apart from a strong binding interaction with each other, relatively little was known about their biochemical function. Recent studies in Drosophila have pinpointed a critical function for the DrosophilaTSC1/TSC2 homologues in the regulation of cell size. Epistasis experiments and a variety of biochemical studies that followed have indicated a critical function for these proteins in the highly conserved PI-3-kinase-Akt-mTOR signalling pathway. PMID- 12556240 TI - The MUC gene family: their role in diagnosis and early detection of pancreatic cancer. AB - The early diagnosis of pancreatic cancer, as well as distinguishing between chronic pancreatitis and malignant pancreatic disease, remains still a clinical problem. Presently, there is no specific tumor marker for diagnosing pancreatic cancer. Mucin-associated marker like CA 19-9 are the most widely available pancreatic cancer tumor marker, but its value as a screening marker is limited by its reduced specificity. Mucins (MUCs) are heavily glycosylated, high molecular weight glycoproteins with an aberrant expression profile in various malignancies. This review considers briefly the potential use of the mucin expression pattern in diagnosis of pancreatic neoplasm. The overview will point out the present knowledge about changes in the mucin gene expression in pancreatic intraepithelial neoplasia (PanINs) as precursor lesions and in pancreatic adenocarcinoma, compared to normal pancreas and chronic pancreatitis and the potential role for differentiating chronic pancreatitis from pancreatic cancer.Furthermore, the potential use of MUCs in the diagnosis and differentiation of intraductal papillary-mucinous neoplasm's (IPMNs) will be discussed. PMID- 12556243 TI - Location-based health information services: a new paradigm in personalised information delivery. AB - Brute health information delivery to various devices can be easily achieved these days, making health information instantly available whenever it is needed and nearly anywhere. However, brute health information delivery risks overloading users with unnecessary information that does not answer their actual needs, and might even act as noise, masking any other useful and relevant information delivered with it. Users' profiles and needs are definitely affected by where they are, and this should be taken into consideration when personalising and delivering information to users in different locations. The main goal of location based health information services is to allow better presentation of the distribution of health and healthcare needs and Internet resources answering them across a geographical area, with the aim to provide users with better support for informed decision-making. Personalised information delivery requires the acquisition of high quality metadata about not only information resources, but also information service users, their geographical location and their devices. Throughout this review, experience from a related online health information service, HealthCyberMap http://healthcybermap.semanticweb.org/, is referred to as a model that can be easily adapted to other similar services. HealthCyberMap is a Web-based directory service of medical/health Internet resources exploring new means to organise and present these resources based on consumer and provider locations, as well as the geographical coverage or scope of indexed resources. The paper also provides a concise review of location-based services, technologies for detecting user location (including IP geolocation), and their potential applications in health and healthcare. PMID- 12556242 TI - The relationship between diabetes and pancreatic cancer. AB - About 80% of pancreatic cancer patients have glucose intolerance or frank diabetes. This observation has led to the following two hypotheses: i. pancreatic cancer causes the associated diabetes and ii. the conditions associated with diabetes promote the development of pancreatic cancer. Evidence supporting both hypotheses has been accumulated in previous studies. This article reviews these studies, especially those that have been conducted recently. PMID- 12556241 TI - Pathways for aberrant angiogenesis in pancreatic cancer. AB - Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease. Although the specific mechanisms that dictate its biological aggressiveness are not clearly established, it is characterized by a variety of molecular alterations as well as by the overexpression of mitogenic and angiogenic growth factors and their receptors. PDACs also express high levels of vascular endothelial growth factor (VEGF). Recent studies indicate that suppression of VEGF expression attenuates pancreatic cancer cell tumorigenicity in a nude mouse model, and that VEGF can exert direct mitogenic effects on some pancreatic cancer cells. These findings suggest that cancer cell derived VEGF promotes pancreatic cancer growth in vivo via a paracrine angiogenic pathway and an autocrine mitogenic pathway, and provide novel opportunities for therapeutic intervention in this deadly disease. PMID- 12556244 TI - The use of interactive graphical maps for browsing medical/health Internet information resources. AB - As online information portals accumulate metadata descriptions of Web resources, it becomes necessary to develop effective ways for visualising and navigating the resultant huge metadata repositories as well as the different semantic relationships and attributes of described Web resources. Graphical maps provide a good method to visualise, understand and navigate a world that is too large and complex to be seen directly like the Web. Several examples of maps designed as a navigational aid for Web resources are presented in this review with an emphasis on maps of medical and health-related resources. The latter include HealthCyberMap maps http://healthcybermap.semanticweb.org/, which can be classified as conceptual information space maps, and the very abstract and geometric Visual Net maps of PubMed http://pubmed.antarcti.ca/start. Information resources can be also organised and navigated based on their geographic attributes. Some of the maps presented in this review use a Kohonen Self Organising Map algorithm, and only HealthCyberMap uses a Geographic Information System to classify Web resource data and render the maps. Maps based on familiar metaphors taken from users' everyday life are much easier to understand. Associative and pictorial map icons that enable instant recognition and comprehension are preferred to geometric ones and are key to successful maps for browsing medical/health Internet information resources. PMID- 12556245 TI - Hypersensitivity vasculitis induced by cefoperazone/sulbactam. AB - BACKGROUND: Cefoperazone has not been reported to cause vasculitic complications before. Here, we report a case of hypersensitivity vasculitis associated with cefoperazone/sulbactam. CASE PRESENTATION: A 13-year-old girl with appendicitis developed hypersensitivity vasculitis on the fifth day of cefoperazone/sulbactam therapy. Hypersensitivity vasculitis resolved gradually after removal of the agent on the seventh day and did not recur. Although hypersensitivity vasculitis has multiple causes, coexistence of hypersensitivity vasculitis and cefoperazone treatment, and the quite resolution of the disease after removal of the drug, strongly favours a causative relationship. CONCLUSION: To our knowledge, this is the first report of a hypersensitivity vasculitis associated with cefoperazone. PMID- 12556247 TI - Reliability, validity and psychometric properties of the Greek translation of the Major Depression Inventory. AB - BACKGROUND: The Major Depression Inventory (MDI) is a brief self-rating scale for the assessment of depression. It is reported to be valid because it is based on the universe of symptoms of DSM-IV and ICD-10 depression. The aim of the current preliminary study was to assess the reliability, validity and psychometric properties of the Greek translation of the MDI. METHODS: 30 depressed patients of mean age 23.41 (+/- 5.77) years, and 68 controls patients of mean age 25.08 (+/- 11.42) years, entered the study. In 18 of them, the instrument was re-applied 1-2 days later and the Translation and Back Translation made. Clinical diagnosis was reached with the use of the SCAN v.2.0 and the International Personality Disorders Examination (IPDE). The Center for Epidemiological Studies-Depression (CES-D) and the Zung Depression Rating Scale (ZDRS) were applied for cross validation purposes. Statistical analysis included ANOVA, the Spearman Product Moment Correlation Coefficient, Principal Components Analysis and the calculation of Cronbach's alpha. RESULTS: Sensitivity and specificity were 0.86 and 0.94, respectively, at 26/27. Cronbach's alpha for the total scale was equal to 0.89. The Spearman's rho between MDI and CES-D was 0.86 and between MDI and ZDRS was 0.76. The factor analysis revealed two factors but the first accounted for 54% of variance while the second only for 9%. The test-retest reliability was excellent (Spearman's rho between 0.53 and 0.96 for individual items and 0.89 for total score). CONCLUSION: The current study provided preliminary evidence concerning the reliability and validity of the Greek translation of the MDI. Its properties are similar to those reported in the international literature, but further research is necessary. PMID- 12556248 TI - A novel high efficiency, low maintenance, hydroponic system for synchronous growth and flowering of Arabidopsis thaliana. AB - BACKGROUND: Arabidopsis thaliana is now the model organism for genetic and molecular plant studies, but growing conditions may still impair the significance and reproducibility of the experimental strategies developed. Besides the use of phytotronic cabinets, controlling plant nutrition may be critical and could be achieved in hydroponics. The availability of such a system would also greatly facilitate studies dealing with root development. However, because of its small size and rosette growth habit, Arabidopsis is hardly grown in standard hydroponic devices and the systems described in the last years are still difficult to transpose at a large scale. Our aim was to design and optimize an up-scalable device that would be adaptable to any experimental conditions. RESULTS: An hydroponic system was designed for Arabidopsis, which is based on two units: a seed-holder and a 1-L tank with its cover. The original agar-containing seed holder allows the plants to grow from sowing to seed set, without transplanting step and with minimal waste. The optimum nitrate supply was determined for vegetative growth, and the flowering response to photoperiod and vernalization was characterized to show the feasibility and reproducibility of experiments extending over the whole life cycle. How this equipment allowed to overcome experimental problems is illustrated by the analysis of developmental effects of nitrate reductase deficiency in nia1nia2 mutants. CONCLUSION: The hydroponic device described in this paper allows to drive small and large scale cultures of homogeneously growing Arabidopsis plants. Its major advantages are its flexibility, easy handling, fast maintenance and low cost. It should be suitable for many experimental purposes. PMID- 12556246 TI - Influence of diabetes mellitus on heart failure risk and outcome. AB - Our aim is to summarize and discuss the recent literature linking diabetes mellitus with heart failure, and to address the issue of the optimal treatment for diabetic patients with heart failure. THE STUDIES LINKING DIABETES MELLITUS (DM) WITH HEART FAILURE (HF) : The prevalence of diabetes mellitus in heart failure populations is close to 20% compared with 4 to 6% in control populations. Epidemiological studies have demonstrated an increased risk of heart failure in diabetics; moreover, in diabetic populations, poor glycemic control has been associated with an increased risk of heart failure. Various mechanisms may link diabetes mellitus to heart failure: firstly, associated comorbidities such as hypertension may play a role; secondly, diabetes accelerates the development of coronary atherosclerosis; thirdly, experimental and clinical studies support the existence of a specific diabetic cardiomyopathy related to microangiopathy, metabolic factors or myocardial fibrosis. Subgroup analyses of randomized trials demonstrate that diabetes is also an important prognostic factor in heart failure. In addition, it has been suggested that the deleterious impact of diabetes may be especially marked in patients with ischemic cardiomyopathy. TREATMENT OF HEART FAILURE IN DIABETIC PATIENTS : The knowledge of the diabetic status may help to define the optimal therapeutic strategy for heart failure patients. Cornerstone treatments such as ACE inhibitors or beta-blockers appear to be uniformly beneficial in diabetic and non diabetic populations. However, in ischemic cardiomyopathy, the choice of the revascularization technique may differ according to diabetic status. Finally, clinical studies are needed to determine whether improved metabolic control might favorably influence the outcome of diabetic heart failure patients. PMID- 12556249 TI - Health care in America: an error-prone system in need of a fix. PMID- 12556251 TI - Assessing need for long-term oxygen therapy: a comparison of conventional evaluation and measures of ambulatory oximetry monitoring. AB - BACKGROUND: Appropriate identification of hypoxic patients with chronic obstructive pulmonary disease (COPD) is important because of the demonstrated survival benefit of long-term oxygen therapy (LTOT) and its associated cost. Resting oxygen saturation (measured via pulse oximetry [S(pO2)]) and lowest exercise S(pO2) (during a 6-min walk test) is the standard method of determining LTOT requirements, but that method does not measure the patient's oxygenation during sleep or activities of daily living. We hypothesized that values obtained via the standard method would correlate poorly with values obtained via ambulatory oximetry monitoring. METHODS: We conducted a prospective, cohort study in an out-patient pulmonary clinic in a tertiary care referral center, with 20 stable COPD patients who were being evaluated for LTOT with conventional evaluation versus 16-24 hours of ambulatory oximetry. RESULTS: The resting S(pO2) did not correlate well with mean ambulatory S(pO2) (r = 0.64) or the percent of monitored time spent with S(pO2) < 88% (r = 0.49). The lowest exercise S(pO2) also did not predict mean ambulatory S(pO2) (r = 0.39) or the percent of monitored time spent with S(pO2) < 88% (r = 0.32). Conventional evaluation overestimated LTOT requirements with 16 of the 20 patients developing an S(pO2) < 88%, most of them with exercise only (ie, most had normal resting S(pO2)). With ambulatory monitoring, however, only 3 of the 16 patients spent > 10% of the monitored time with S(pO2) < 88%. CONCLUSION: There was a poor relationship between the conventional oxygenation assessment method and continuous ambulatory oximetry during LTOT screening with COPD patients. PMID- 12556250 TI - Missed bronchodilator medication treatments in respiratory therapy: frequency and underlying causes. AB - BACKGROUND: In the context of increasing attention to medical errors, missed therapies have become a subject of focus both for optimizing clinical care and for assuring appropriate institutional performance during external review by accrediting bodies. Because the issue of missed treatments in respiratory therapy has received little attention to date, we undertook to describe the frequency and causes of missed respiratory therapy bronchodilator medication treatments at the Cleveland Clinic Hospital. METHODS: Between August 2000 and August 2001, using a respiratory therapy management information system, we recorded the number of respiratory therapy bronchodilator medication treatments ordered and delivered (via small-volume nebulizer and metered-dose inhaler) and the reason(s) for each missed treatment. RESULTS: Over the 12-month study interval 113,554 bronchodilator medication treatments (74,921 via small-volume nebulizer and 38,633 via metered-dose inhaler) were ordered. Overall, 4,012 medication treatments were missed (3.5% of the total), with variation by month ranging from 2.0% to 5.0%. The commonest reason for failure to administer the ordered bronchodilator treatment was the patient being out of the room at the time of the therapist's visit, which accounted for nearly one third of missed therapies. Next most common was the patient refusing treatment (24.6%), followed by the patient being unavailable because of ongoing activities or therapy (eg, physical therapy or a medical procedure). The least common reason was the respiratory therapist being called away to administer therapy to another patient (1.4%). CONCLUSIONS: Overall, the frequency of missed bronchodilator treatments was relatively low in this series. The next steps include developing strategies to lower the frequency of missed treatments, so as to optimize the allocation of respiratory therapy services, and studying the clinical consequences of missed therapies. PMID- 12556252 TI - A comparison of the OxyArm oxygen delivery device and standard nasal cannulae in chronic obstructive pulmonary disease patients. AB - OBJECTIVE: Compare the performance of the OxyArm to that of nasal cannulae in the delivery of supplemental oxygen to patients with chronic obstructive pulmonary disease. METHODS: We tested various oxygen flows with 10 chronic obstructive pulmonary disease patients who were receiving prescribed supplemental oxygen. Blood oxygen saturation (measured via pulse oximetry [S(pO2)]), was measured with each device, and mean data were compared with paired, 2-sample tests. RESULTS: Mean S(pO2) was equivalent with OxyArm and nasal cannulae for 7 of the 10 subjects, over a range of oxygen flows (2-5 L/min). Mean S(pO2) was higher with the OxyArm with 2 subjects and lower with 1 subject (p < 0.01). CONCLUSION: The OxyArm maintained stable S(pO2) over the range of oxygen flows studied and at levels equivalent to those maintained by nasal cannulae in 9 of the 10 subjects. The OxyArm does not contact the face and may be ideal for patients on long-term home oxygen therapy. PMID- 12556253 TI - Performance comparison of two oscillating positive expiratory pressure devices: Acapella versus Flutter. AB - BACKGROUND: Oscillatory positive expiratory pressure (PEP) with the Flutter device facilitates secretion removal. In the Flutter a steel ball vibrates inside a cone, causing air flow vibration. A new device, the Acapella, uses a counterweighted plug and magnet to create air flow oscillation. The Acapella comes in 2 models: one for patients with expiratory flow > or = 15 L/min and one for < or = 15 L/min. We hypothesized that the Acapella and Flutter would produce similar mean PEP, oscillatory pressure amplitude, and frequency over a clinically relevant range of flows. METHODS: We measured oscillatory amplitude, PEP, and frequency. Values for frequency, peak, trough, and mean pressure were recorded automatically every 3 seconds at flows of 5, 10, 15, 20, 25, and 30 L/min. The pressure waveform for 1 second was also graphically displayed and recorded. The devices were adjusted to give low, medium, and high mean expiratory pressure (Flutter angle at 0, 20, and 40 degrees; Acapella by dial setting). Data were analyzed by 2-way repeated measures analysis of variance, and differences were considered significant when p was < 0.05. RESULTS: There were statistically significant differences between the devices for mean pressure, pressure amplitude, and frequency, for all experimental conditions. However, the differences were relatively small and may not be clinically important. Both devices produced similar pressure waveforms at the medium flows. At 5 L/min the Acapella produced a more stable waveform, with a lower frequency, higher amplitude, and a slightly wider range of PEP than the Flutter. CONCLUSIONS: Acapella and Flutter have similar performance characteristics. Acapella's performance is not gravity-dependent (ie, dependent on device orientation) and may be easier to use for some patients, particularly at low expiratory flows. PMID- 12556254 TI - Development of an affordable data collection, reporting, and analysis system. AB - Available professionally designed systems for recording, storing, retrieving, and analyzing patient data (such as treatments delivered, laboratory and pulmonary function test results, ventilator settings, and patient assessments) are prohibitively expensive to many hospitals and clinics. At Tufts-New England Medical Center we designed and implemented a computerized patient data management system that uses relatively inexpensive "personal digital assistants" and inexpensive, commonly-available software. Compared to the pen-and-paper flow sheet system we previously used, our computerized system saves time and money, is accurate and user-friendly, and allows data retrieval and analysis that can improve clinical practice and efficiency. We believe our experience could benefit other institutions. PMID- 12556255 TI - Diagnosis of a space-occupying lesion of the lung by pulmonary function tests. PMID- 12556256 TI - Airway clearance in cystic fibrosis. PMID- 12556257 TI - Bacteremia, infection, and antibiotic choices. PMID- 12556258 TI - A comparison of the therapeutic effectiveness of and preference for postural drainage and percussion, intrapulmonary percussive ventilation, and high frequency chest wall compression in hospitalized cystic fibrosis patients. AB - INTRODUCTION: Cystic fibrosis (CF) patients have abnormally viscid bronchial secretions that cause airway obstruction, inflammation, and infection that leads to lung damage. To enhance airway clearance and reduce airway obstruction, daily bronchopulmonary hygiene therapy is considered essential. OBJECTIVE: Compare the effectiveness of and patient preferences regarding 3 airway clearance methods: postural drainage and percussion (PD&P), intrapulmonary percussive ventilation (IPV), and high-frequency chest wall compression (HFCWC). METHODS: The participants were hospitalized CF patients >or= 12 years old. Effectiveness was evaluated by measuring the wet and dry weights of sputum obtained with each method. In random order, each patient received 2 consecutive days of each therapy, delivered 3 times daily for 30 minutes. Sputum was collected during and for 15 minutes after each treatment, weighed wet, then dried and weighed again. Participants rated their preferences using a Likert-type scale. Mean weights and preferences were compared using analysis of variance with repeated measures. Patient preferences were compared using Freidman's test. RESULTS: Twenty-four patients were studied. The mean +/- SD wet sputum weights were 5.53 +/- 5.69 g with PD&P, 6.84 +/- 5.41 g with IPV, and 4.77 +/- 3.29 g with HFCWC. The mean wet sputum weights differed significantly (p = 0.035). Wet sputum weights from IPV were significantly greater than those from HFCWC (p < 0.05). The mean dry sputum weights were not significantly different. With regard to overall preference and to the subcomponents of preference, none of the 3 methods was preferred over the others. CONCLUSIONS: HFCWC and IPV are at least as effective as vigorous, professionally administered PD&P for hospitalized CF patients, and the 3 modalities were equally acceptable to them. A hospitalized CF patient should try each therapy and choose his or her preferred modality. PMID- 12556259 TI - Effects of a simple protocol on infective complications in intensive care unit patients undergoing percutaneous dilatational tracheostomy. AB - In our intensive care unit we monitored infection in 228 patients who underwent percutaneous dilatational tracheostomy (PDT). In the first phase of the study 128 PDTs were performed during a 33-month period and there were 41 infection complications (nosocomial pneumonia, bacteremia with sepsis, and septic shock) in the perioperative period (immediately prior to and for 5 days after PDT). A significant risk factor among patients with nosocomial pneumonia was empirical administration of inappropriate antibiotics, compared to appropriate antibiotics (34% versus 4%, p < 0.001). In the second phase of the study (a 30-month period), a simple antibiotics protocol was prospectively applied to 100 PDT patients. The protocol virtually eliminated inappropriate antibiotic drug use immediately prior to PDT and contributed to a significant reduction in perioperative infective complications (pre-protocol 32% versus protocol 11%, p < 0.001). PMID- 12556260 TI - Spontaneously breathing lung model comparison of work of breathing between automatic tube compensation and pressure support. AB - INTRODUCTION: Automatic tube compensation (ATC), a new ventilation mode that compensates for the work of breathing imposed by endotracheal tube resistance, recently became commercially available. METHODS: We conducted a laboratory study with a lung model and a Nellcor Puritan Bennett 840 ventilator to compare ATC and pressure-support ventilation (PSV). A bellows-in-a-box lung model simulated spontaneous breathing with the following settings: respiratory rate 10 breaths/min, inspiratory time 1.0 s, peak inspiratory flow 60 L/min without connecting to the ventilator and endotracheal tube (ETT). At each ETT size (5, 6, 7, 8 and 9 mm inner diameter) 100% ATC was compared with pressure support (PS) of 0, 2, 4, 6, 8, and 10 cm H(2)O at positive end-expiratory pressure (PEEP) of 0 and 5 cm H(2)O. The negative deflection (PI) of the "alveolar" pressure (ie, pressure inside the bellows, P(alv)) and the delay time were measured. The PI and total pressure-time product (PTP(tot)) integrated from P(alv) were analyzed. PTP(tot) was subdivided into PTP(trig) (the PTP from the beginning of inspiration to the minimum P(alv)) and PTP(supp) (the PTP from the minimum P(alv) to the return to baseline P(alv)). RESULTS: At PEEP of 0 cm H(2)O: with ETTs of 5, 6, and 7 mm the PI values with ATC corresponded to PS of 0-4 cm H(2)O; with the 8-mm ETT the PI values corresponded to PS of 0 cm H(2)O; with the 9-mm ETT the PI values corresponded to PS of 0-2 cm H(2)O. At PEEP of 5 cm H(2)O, with all ETT sizes the PI values corresponded to PS of 0 cm H(2)O. PTP(tot) and PTP(supp) of ATC corresponded to: PS of 2-4 cm H(2)O with the 5-mm ETT; PS of 2 cm H(2)O with the 6-mm ETT; PS of 0-2 cm H(2)O with the 7-mm ETT; and PS of 0 cm H(2)O with the 8- and 9-mm ETTs, at PEEP of 0 cm H(2)O. PEEP of 5 cm H(2)O was not tested for PTP. PTP(trig) with ATC showed comparable or greater values with each size of ETT. CONCLUSIONS: ATC with a Nellcor Puritan Bennett 840 ventilator provided inspiratory ventilatory support corresponding to PS of or= 50,000 dollars/year. Participation rates averaged 62%. MAIN OUTCOME MEASURES: The major domain investigated was consumer support for sustainably produced foods. Demographics, beliefs, attitudes, subjective norm, and self-identity and perceived behavioral control were evaluated as predictors of intention to purchase them. ANALYSIS: Descriptive statistics, independent t tests, one-way analysis of variance, Pearson product moment correlation coefficients, and stepwise multiple regression analyses (P <.05). RESULTS: Consumers were supportive of sustainably produced foods but not highly confident in their ability to purchase them. Independent predictors of intention to purchase them included attitudes, beliefs, perceived behavioral control, subjective norm, past buying behavior, and marital status. CONCLUSIONS AND IMPLICATIONS: Beliefs, attitudes, and confidence level may influence intention to purchase sustainably produced foods. Nutrition educators could increase consumers' awareness of sustainably produced foods by understanding their beliefs, attitudes, and confidence levels. PMID- 12556269 TI - Do food-related experiences in the first 2 years of life predict dietary variety in school-aged children? AB - OBJECTIVE: To determine if food-related experiences in the first 2 years of life predict dietary variety in school-aged children. DESIGN/SETTING: Child/mother pairs were interviewed 7 or 8 times when children were 2 to 24 months using a randomized incomplete block design to schedule interviews. Each child/mother pair was interviewed when the child was ages 6, 7, and 8 years. PARTICIPANTS: Child/mother pairs (n = 70) were continuous participants in the longitudinal study. MAIN OUTCOME MEASURES: Dependent variables were children's vegetable and fruit dietary variety, assessed from 3 days of dietary data at ages 6, 7, and 8 years. Independent variables from the first 2 years of life were selected from the longitudinal data set. ANALYSES: General linear models. Adjustments for age that vegetables (or fruits) were introduced in the diet. RESULTS: Vegetable variety in the school-aged child was predicted by mother's vegetable preferences, R2 =.084. Fruit variety in the school-aged child was predicted by breast-feeding duration and either early fruit variety (R2 =.254) or fruit exposure (R2 =.246). CONCLUSIONS/IMPLICATIONS: Nutrition education messages for mothers should emphasize the importance of early food-related experiences to school-aged children's acceptance of a variety of vegetables and fruits. PMID- 12556271 TI - Coping strategies and nutrition education needs among food pantry users. AB - In response to unexpectedly high rates of household food insecurity in Washington State, focus groups were conducted with food pantry users in nine locations across the state to explore access and barriers to food, strategies for coping with food insecurity, and nutrition education needs. All respondents had children and spoke English. Participants reported household food insecurity and concerns about feeding children and getting through the month. Stretching food dollars was the most requested topic for nutrition education. Nutrition educators need to be responsive to people's educational needs and the emotional demands associated with food insecurity at the program and community levels. PMID- 12556272 TI - Strategies to motivate students in online learning environments. AB - We are happy to present the following article as a resource for readers developing or refining online courses. Much work remains to be done to identify and evaluate the relative merits of potential theoretical models for online nutrition instruction. Progress will depend on finding ways to conduct objective appraisals of theory-driven courses in university settings, where randomization is often impossible and courses are often refined in real time to meet students' immediate needs or to take advantage of rapidly evolving technology. Meanwhile, we hope that the concepts and observations described in this Special Article stimulate consideration of the motivational aspects of online courses in practice, research, and institutional policy making. PMID- 12556273 TI - Kentucky state fair cooking school: connecting farm to table. PMID- 12556274 TI - Let's move, learn, and have fun! PMID- 12556275 TI - Routine vaccines across the life span, 2003. AB - This review of the recommendations for routine vaccines includes those used on the Recommended Childhood Immunization Schedule and the Recommended Adult Immunization Schedule. The rationale is provided for recent changes, such as encouraging influenza vaccination of healthy children aged 6 through 23 months and recommending that annual influenza vaccination begin at age 50. The risks and benefits of vaccination are discussed, including disease burden, rationale for vaccination, efficacy, adverse reactions, and recommendations. Pictures of vaccine-preventable diseases are included. PMID- 12556276 TI - Vaccines for persons at high risk due to medical conditions, occupation, environment, or lifestyle, 2003. AB - The safety and efficacy of current vaccines are reviewed for high-risk populations, such as those with underlying medical conditions or occupational or lifestyle circumstances. The morbidity and mortality from vaccine-preventable diseases are high among persons with underlying medical conditions; thus, influenza and pneumococcal polysaccharide vaccines are recommended for those with cardiac disease, diabetes mellitus, or chronic obstructive pulmonary disease. For the same reasons, influenza vaccine is recommended for pregnant women and for persons with asthma. Health-care workers are at risk for acquiring and transmitting hepatitis B, measles, and influenza; hence, vaccination against these diseases is recommended. PMID- 12556277 TI - Vaccine schedules and procedures, 2003. AB - Recent, rapid changes in recommendations regarding vaccination represent challenges for busy clinicians who need to be familiar with new developments. This article reviews the current age-based recommendations that are fundamental to the United States vaccine program, as well as current recommendations regarding contraindications, precautions, administration techniques, and storage procedures for routinely administered vaccines. The 2003 Recommended Childhood Immunization Schedule, the 2002-2003 Recommended Adult Immunization Schedule, contraindications for immunization, and general guidelines on immunization procedures are presented. Recent changes in recommendations for children are instituting thimerosal-free hepatitis B vaccine as soon after birth as possible (before hospital discharge) and encouraging immunization of all children aged 6 to 23 months against influenza. PMID- 12556278 TI - Addressing immunization barriers, benefits, and risks. AB - Immunization rates in the United States still fall short of the Healthy People 2010 goals for children and adults. To improve rates of immunization, physicians need to understand and address barriers to immunization, including fragmented health-care delivery, missed opportunities to vaccinate, and the patient's fear of adverse reactions. This article addresses these issues and suggests strategies by which rates can be improved, such as patient reminders, standing orders, and assessment of and feedback on practitioner performance. Additionally, it provides suggestions to help physicians better communicate vaccine risks and benefits to their patients, potentially affecting an individual's acceptance of those risks. It describes the appropriate use of materials such as the Vaccine Information Statements. Physicians should also be prepared to answer patients' questions about alleged or controversial vaccine adverse events. PMID- 12556279 TI - Vaccines and bioterrorism: smallpox and anthrax. AB - Because of the success of vaccination and the ring strategy in eradicating smallpox from the world, smallpox vaccine has not been recommended for the United States civilian populations for decades. Given the low but possible threat of bioterrorism, smallpox vaccination is now recommended for those teams investigating potential smallpox cases and for selected personnel of acute-care hospitals who would be needed to care for victims in the event of a terrorist attack. Treatment and post-exposure prophylaxis for anthrax are ciprofloxacin or doxycycline. Anthrax vaccine alone is not effective for post-exposure prevention of anthrax; vaccination is accompanied by 60 days of antibiotic therapy. In addition to military use, anthrax vaccine is recommended for pre-exposure use in those persons whose work involves repeated exposure to Bacillus anthracis spores. PMID- 12556280 TI - Explaining gender segregation. AB - Occupational gender segregation--the tendency for women and men to work in different occupations--is an important feature of all societies, and particularly the wealthy industrialized ones. To understand this segregation, and to explain its significance, we need to distinguish between vertical segregation entailing inequality and horizontal segregation representing difference without inequality, with overall segregation being the resultant of these components. Three major theoretical approaches to understanding occupational gender segregation are examined: human capital/rational choice, patriarchy, and preference theories. All are found to be inadequate; they tend to confuse overall segregation with its vertical component, and each entails a number of other faults. It is generally assumed or implied that greater empowerment of women would reduce gender segregation. This is the reverse of what actually happens; in countries where the degree of women's empowerment is greater, the level of gender segregation is also greater. An alternative theoretical approach based on processes of social reproduction is shown to be more useful. PMID- 12556281 TI - Employment, flexible working and the family. AB - This paper assesses some of the implications of one of the major social changes to have taken place in the West during the second half of the twentieth century- that is, the increased employment of women, together with normative changes in gender relations and in women's expectations. These changes have been linked to an increase in individualism, which itself is associated with the transcendence of 'first modernity'. Thus it is suggested that new approaches to social analysis are required (Beck). Here it is argued that, rather than develop completely new approaches in order to grasp the changes that are under way, the 'economic' and the 'social' (that is, employment and the family) should be seen as intertwined, rather than approached as separate phenomena. Past debates in feminism, changes in the family, and flexible employment are critically examined. The growing tensions between employment and family life are discussed. It is argued that these changes are associated with the intensification of capitalist development, rather than reflecting a fundamental transformation of society. Existing approaches to the analysis of social change, including Polanyi's analysis of the development of 'counter-movements' against the 'self-regulating' market, will, therefore, still be relevant to our enquiries. In the concluding section, a programme of research that would examine these changes is outlined. PMID- 12556282 TI - Reflections on the meritocracy debate in Britain: a response to Richard Breen and John Goldthorpe. AB - A paper by Breen and Goldthorpe recently claimed to have exposed 'fatal flaws' in my work on meritocracy in Britain. This paper responds to their criticisms. The results of their re-analysis of the NCDS data set are shown to be consistent with my earlier findings and arguments. Furthermore, re-running some of my earlier models using measures that they favour and a method designed to privilege their position, the results once again demonstrate that, while class origins have some effect on class destinations (in particular, for those born into the middle class), ability and effort exert a much greater effect. Based on these results, the paper identifies three core propositions about meritocracy in Britain on which all parties to this debate should now be able to agree. PMID- 12556283 TI - Merit, mobility and method: another reply to Saunders. PMID- 12556284 TI - Ideal types, stereotypes and classes. AB - Several recent articles seeking to validate the Goldthorpe class schema have implicitly raised some interesting questions about the relation between analytical techniques and theory. The favoured technique of latent class analysis actually brings this variety of class analysis close to the much older tradition of ideal-type analysis. In doing so, it helps to illustrate a much wider problem that arises in any conceptualization in terms of classes. The paper seeks to show how, just as in popular discourse around 'class', stereotypes are used to simplify and summarize a complex set of related characteristics, so similarly ideal types, as used in class analysis, are an over-simplification. As elsewhere in social science, the danger of using them is that they serve to protect a theoretical concept from empirical refutation. An alternative 'research programme' to that of class analysis is presented, based on measures of stratification arrangements that are more closely linked to the reproducing structure of inequalities. PMID- 12556285 TI - Higher education and civic engagement. AB - This paper focuses on the relationship between social engagement, particularly civic engagement, and education. It is well known that more highly educated people are more likely to engage in voluntary work in formalized settings. It has been difficult to disentangle the effect of higher education from that of family origin and occupational socialization. This paper examines the effects of tertiary education on the social and civic engagement of young people, using the British Household Panel Study. The social and civic activity of young people is observed in their late teens, before entering the labour market or tertiary education, and compared with that of the same young people in their early 20s, after completing tertiary education courses or gaining labour market experience. It was found that the social and civic engagement of young people who would enter higher education was higher in their late teens than that of their peers who did not enter. However, higher education had a small additional effect on civic engagement, for both young and mature students. The children of professionals were the social grouping most likely to be involved in civic activities. The relationship of higher education, professional occupations and family socialization is discussed. PMID- 12556286 TI - Culture, the 'sick role' and the consumption of health. AB - This paper revisits Parsons's conception of the 'sick role' and examines the relevance of his writings on the cultural understanding of sickness to the consumption of health in the contemporary era. In terms of current developments, I focus on the development of pro-active approaches towards the healthy body, and the growth of 'information rich' consumers of health care. These have become prominent themes in sociology, and while Parsons's writings are usually viewed as anachronistic I argue they remain highly pertinent to understanding the emergence of informed, body conscious lay people. If Parsons's analysis of health is more relevant to current circumstances than many critics assume, however, it is not unproblematic. The residual categories associated with the sick role obscure the continued utility of his work on the general cultural values informing health care. It is Parsons's analysis of these values, I suggest, that needs rescuing from restricted understandings of the sick role and highlighting as an important resource for contemporary theorists. PMID- 12556287 TI - Positivism in sociological research: USA and UK (1966-1990). AB - Positivism has been declared dead in sociological theory circles, yet questions remain as to its viability among researchers. The authors present diagnostic evidence about positivism in sociological practice through a content analysis of journal articles published in the late 1960s and the late 1980s in the sociological journals of the USA and Britain. Using an index based on seven elements of positivism that were characteristic of the 'theory construction' movement of the late 1960s, the authors find evidence of the effects of time and nation on the use of positivism. Disaggregation of the index reveals that most of the observed change is associated with the elements of 'instrumental' positivism, particularly statistics. The results raise questions about the relationship between theory and research and about sociologists' philosophies of science. PMID- 12556288 TI - Complexity and practical knowledge in the social sciences: a comment on Stehr and Grundmann. AB - This paper takes issue with Stehr and Grundmann's argument in this journal that sociology's poor record of contribution to practical knowledge results from sociologists' misguided attachment to the idea that social phenomena are peculiarly complex, and so peculiarly resistant to efforts at practical knowledge formation. Stehr and Grundmann develop their thesis by contrasting sociology to economics, claiming the latter's superior methods and disciplinary cultures enable it to contribute more successfully to solving practical problems. We argue that Stehr and Grundmann's comparison of the relative success of economics and sociology as policy sciences underestimates the extent to which the complexity argument has been destructive of economics' capacity to contribute practical knowledge, and overestimates the importance of disciplinary methods and cultures in explaining the use of economics in policy making. Further, we argue that the 'complexity argument' can contribute to better policy making, by virtue of its capacity to highlight the dangers of simplistic or fundamentalist solutions to social problems with complex causes. PMID- 12556289 TI - Global anti-corporate struggle: a preliminary analysis. AB - This paper offers a preliminary analysis of the recent wave of anti-corporate protest that has swept across numerous countries throughout the world. In the first part of the paper the social structure of this phenomenon is examined. Specifically, it is argued that it should be understood as a 'field', in Pierre Bourdieu's sense of the term. In the second part of the paper the factors which account for the emergence of this field are explored, using a 'value-added' model which focuses upon the interplay of strains, situational definitions, focal events, opportunities and the circulation of protest relevant resources. The paper is conceived as a preliminary analysis which outlines a framework and draws out important themes. It is not offered as a complete account but rather as a basis from which more specific and focused studies might stem. PMID- 12556291 TI - Diagnosis of congenital nephrotic syndrome: a clinical and a pathologic challenge. AB - The diagnosis of congenital nephrotic syndrome (NS) is a challenge both for clinicians and for pathologists. We observed three cases in a series of 50 children with NS nonresponsive to therapy, corresponding to one case each of minimal change disease, Finnish-type glomerulopathy, and diffuse mesangial sclerosis--two histopathologic studies were performed in each case. The age at presentation did not predict the diagnosis nor the prognosis: The NS presented at 7 months of age in the patient with diffuse mesangial sclerosis, but it was present at birth in the patient with minimal change disease. In these 2 patients the final diagnosis was made with the first renal biopsy. Conversely, in the patient with Finnish-type glomerulopathy, the diagnosis was only possible in the repeat biopsy, as the early pathologic changes were nonspecific. This study shows the essential role of the renal biopsy in determining the etiologic diagnosis and prognosis in patients with congenital nephrotic syndrome. congenital nephrotic syndrome diffuse mesangial sclerosis Finnish-type glomerulopathy PMID- 12556292 TI - SIALYL-Tn antigen distribution in Helicobacter pylori chronic gastritis in children: an immunohistochemical study. AB - Sialyl-Tn antigen (STn) is a mucin-type carbohydrate normally present in goblet cells of small and large bowel. STn expression has been demonstrated to occur in complete and incomplete intestinal metaplasia as well as in many carcinomas but in no normal gastric cell. The aim of our present study was to evaluate the distribution of STn in Helicobacter pylori chronic gastritis (HpCG) of pediatric patients. Eighteen gastric biopsies from 15 children (mean age: 11.5 years) with HpCG, 9 gastric biopsies from 9 children without H. pylori infection, and 1 heterotopic gastric mucosa in Meckel's diverticulum were immunostained using the anti-STn antibody STn1 (18/18), NCL-MUC-1 (7/18), and NCL-MUC-2 (18/18) antibodies. Also, sulfated mucosubstances were investigated with the Alcian Blue Periodic Acid Schiff (AB-PAS), pH 1.0 stain. Although with different intensity (weak in 5/18, moderate 9/18, and intense 4/18) all cases with HpCG exhibited STn immunoreactivity. The expression of STn was found to be located mainly to the supranuclear region of the epithelial cells at the foveolae and glandular necks, with occasional cells showing diffuse cytoplasmic staining. When reactivity was intense, it was for the most part found in the cells at the neck of the glands. The mucus out of the luminal border above the positive cells was usually also stained. MUC-1 was negative (2/7) or weakly positive (5/7) in a few surface mucous cells. MUC-2 was negative (16/18) or occasionally detected in some foveolar and surface cells (2/18). AB-PAS pH 1.0 revealed the presence of sulfomucins in the cytoplasm of isolated cells of gastric pits and glands of most cases (11/15). None of these findings was observed in the control group. We conclude that STn can be identified in gastric cells of pediatric patients with HpCG and that this does not correlate with other mucosubtances markers. The findings could indicate that minimal intestinal metaplasia takes place in children with HpCG. PMID- 12556293 TI - Effects of oral genistein in mice. AB - In cell culture systems, genistein, a soy-derived isoflavone with chemopreventive and estrogenic effects, enhances cAMP-dependent activation of the most common cystic fibrosis-causing mutation, deltaF508-CFTR, by as much as 20-fold. DeltaF508-CFTR is present in the apical membrane at far lower levels than wild type CFTR. If genistein can enhance cyclic AMP-dependent activity in vivo, the presence of deltaF508-CFTR, at even a few percent of wild-type levels, might permit genistein to be of therapeutic benefit to cystic fibrosis patients with this mutation. Before determining if oral genistein would be of benefit in mice with a deltaF508 mutation in the murine CFTR gene, a maximal dose of oral genistein with minimal side effects needed to be established. Accordingly, C57Bl/6 mice pups were randomly weaned onto soy-free diet, AIN-76, containing between 0 and 1.0 g/kg genistein and allowed to feed ad libitum for 3 weeks. Genistein had no significant effects on growth rates of either male or female mice. Histology of the lung, heart, kidney, liver, and intestine revealed no significant genistein-dependent changes in morphology. When mice on a 1.0 g/kg of genistein diet were sacrificed in the morning, the mean level of serum genistein was 1.4+/-0.2 micro moles/L. Serum genistein increased during the daylight hours reaching a maximum of 7.5+/-0.6 micro moles/L in the early evening. Our results demonstrate that dietary genistein is not inhibitory to growth or caloric intake and up to 1.0 g/kg ad libitum genistein causes no significant organ specific abnormalities. PMID- 12556294 TI - DNA content and proliferative activity in pediatric genitourinary rhabdomyosarcoma. AB - Most of the studies have reported that cases of pediatric rhabdomyosarcoma (RMS) with hyperdiploid DNA and low cellular proliferative activity have better outcomes. The aim of our study was to evaluate the possible clinical relevance of DNA ploidy and proliferative activity in childhood genitourinary RMS. Twelve childhood genitourinary RMS cases were reexamined histologically and correlated with clinical features and patient survival. DNA analysis was performed on cytospin single-cell preparations obtained from the paraffin-embedded tissue blocks. MIB-1 was the proliferative marker used on paraffin sections. All patients were male with a mean age at diagnosis of 65 months. There were 5 tumors on the bladder, 5 on the paratesticular region, and 2 on the prostate. All cases were treated with multimodality therapy using the protocols proposed by the International Society of Pediatric Oncology. The following subtypes of RMS were recognized: embryonal (n=6), spindle cell (n=4), and botryoid (n=2). DNA hyperdiploid was detected in 11 tumors (92%) and high MIB-1 index (>19%) in 4 cases (33%). Now, 10 patients are alive without evidence of disease and 2 patients are alive with evidence of disease. Our data suggested that childhood RMS of the genitourinary tract are preponderantly DNA hyperdiploid and have low cellular proliferative activity. It is also interesting that 83% of our genitourinary pediatric RMS patients are alive. As consequence of this study, we propose that DNA content and proliferative activity is a useful method to supplement findings in children with RMS. PMID- 12556295 TI - Late onset ornithine transcarbamylase deficiency: a case report. AB - Ornithine transcarbamylase deficiency, an X-linked disorder, is the most common inherited urea cycle defect. Previous reports have documented the existence of several different mutations that can, partly at least, explain the phenotypic variability of the disorder. We describe the only male with T343K mutation, which also is present in his mother. We underline the disorientation of the beginning of clinical presentation; the patient became ill when fruits were added to his diet. PMID- 12556296 TI - The isoprenoid pathway and the pathogenesis of Reye's syndrome. AB - The isoprenoid pathway produces three key metabolites: endogenous digoxin (regulator of neurotransmitter uptake), dolichol, and ubiquinone (free radical scavenger). Because a mitochondrial dysfunction has been described in Reye's syndrome, we thought it pertinent to assess the pathway in this disease. Since endogenous digoxin can regulate neurotransmitter transport, the pathway also was assessed in patients with right hemispheric, left hemispheric, and bihemispheric dominance to find out the role of hemispheric dominance in its pathogenesis. The plasma/serum activity of HMG CoA reductase, magnesium, digoxin, dolichol, ubiquinone, tryptophan/tyrosine catabolic patterns, free radical, and lipid levels as well as (red blood cell) RBC Na(+)-K(+) ATPase activity were measured in the above mentioned groups. RESULTS: In the patient group as well as in individuals with right hemispheric dominance similar patterns were obtained. There was elevated digoxin and dolichol levels with low levels of ubiquinone in patients with Reye's syndrome as well as in those with right hemispheric dominance. The serum magnesium and RBC Na(+)-K(+) ATPase activity were reduced. There also was an increase in tryptophan catabolites and a reduction in tyrosine catabolites as well as increased free radical levels. Reye's syndrome is associated with an upregulated isoprenoid pathway, elevated hypothalamic digoxin secretion, and right hemispheric chemical dominance. PMID- 12556297 TI - Pathology teach and tell: melanotic ependymoma. PMID- 12556298 TI - Pathology teach and tell: neonatal ELS. PMID- 12556299 TI - Pathology teach and tell: intravascular endothelial hyperplasia forming colonic polyps. PMID- 12556300 TI - Letter to the editor--multifocal myocardial apoptoses: distinctive cardiac lesions in cystic fibrosis and other diseases. PMID- 12556326 TI - Quantitative comparisons of cancer induction in humans by internally deposited radionuclides and external radiation. AB - PURPOSE: To compare quantitative estimates of lifetime cancer risk in humans for exposures to internally deposited radionuclides and external radiation. To assess the possibility that risks from radionuclide exposures may be underestimated. MATERIALS AND METHODS: Risk estimates following internal exposures can be made for a small number of alpha-particle-emitting nuclides. (1) Lung cancer in underground miners exposed by inhalation to radon-222 gas and its short-lived progeny. Studies of residential (222)Rn exposure are generally consistent with predictions from the miner studies. (2) Liver cancer and leukaemia in patients given intravascular injections of Thorotrast, a thorium-232 oxide preparation that concentrates in liver, spleen and bone marrow. (3) Bone cancer in patients given injections of radium-224, and in workers exposed occupationally to (226)Ra and (228)Ra, mainly by ingestion. (4) Lung cancer in Mayak workers exposed to plutonium-239, mainly by inhalation. Liver and bone cancers were also seen, but the dosimetry is not yet sufficiently good enough to provide quantitative estimates of risks. Comparisons can be made between risk estimates for radiation induced cancer derived for radionuclide exposure and those derived for the A-bomb survivors, exposed mainly to low-LET (linear energy transfer) external radiation. Data from animal studies, using dogs and rodents, allow comparisons of cancer induction by a range of alpha- and beta-/gamma-emitting radionuclides. They provide information on relative biological effectiveness (RBE), dose-response relationships, dose-rate effects and the location of target cells for different malignancies. RESULTS: For lung and liver cancer, the estimated values of risk per Sv for internal exposure, assuming an RBE for alpha-particles of 20, are reasonably consistent with estimates for external exposure to low-LET radiation. This also applies to bone cancer when risk is calculated on the basis of average bone dose, but consideration of dose to target cells on bone surfaces suggests a low RBE for alpha-particles. Similarly, for leukaemia, the comparison of risks from alpha-irradiation ((232)Th and progeny) and external radiation suggest a low alpha RBE; this conclusion is supported by animal data. Risk estimates for internal exposure are dependent on the assumptions made in calculating dose. Account is taken of the distribution of radionuclides within tissues and the distribution of target cells for cancer induction. For the lungs and liver, the available human and animal data provide support for current assumptions. However, for bone cancer and leukaemia, it may be that changes are required. Bone cancer risk may be best assessed by calculating dose to a 50 micro m layer of marrow adjacent to endosteal (inner) bone surfaces rather than to a single 10 micro m cell layer as currently assumed. Target cells for leukaemia may be concentrated towards the centre of marrow cavities so that the risk of leukaemia from bone seeking radionuclides, particularly alpha emitters, may be overestimated by the current assumption of uniform distribution of target cells throughout red bone marrow. CONCLUSIONS: The lifetime risk estimates considered here for exposure to internally deposited radionuclides and to external radiation are subject to uncertainties, arising from the dosimetric assumptions made, from the quality of cancer incidence and mortality data and from aspects of risk modelling; including variations in baseline rates between populations for some cancer types. Bearing in mind such uncertainties, comparisons of risk estimates for internal emitters and external radiation show good agreement for lung and liver cancers. For leukaemia, the available data suggest that the assumption of an alpha-particle RBE of 20 can result in overestimates of risk. For bone cancer, it also appears that current assumptions will overestimate risks from alpha-particle-emitting nuclides, particularly at low doses. PMID- 12556327 TI - Radiation-induced genomic instability and bystander effects: related inflammatory type responses to radiation-induced stress and injury? A review. AB - PURPOSE: To review studies of radiation responses in the haemopoietic system in the context of radiation-induced genomic instability, bystander effects and inflammatory-type processes. RESULTS: There is considerable evidence that cells that themselves are not exposed to ionizing radiation but are the progeny of cells irradiated many cell divisions previously may express a high frequency of gene mutations, chromosomal aberrations and cell death. These effects are collectively known as radiation-induced genomic instability. A second untargeted effect results in non-irradiated cells exhibiting responses typically associated with direct radiation exposure but occurs as a consequence of contact with irradiated cells or by receiving soluble signals from irradiated cells. These effects are collectively known as radiation-induced bystander effects. Reported effects include increases or decreases in damage-inducible and stress-related proteins; increases or decreases in reactive oxygen species, cell death or cell proliferation, and induction of mutations and chromosome aberrations. This array of responses is reminiscent of effects mediated by cytokines and other similar regulatory factors that may involve, but do not necessarily require, gap junction mediated transfer, have multiple inducers and a variety of context-dependent consequences in different cell systems. That chromosomal instability in haemopoietic cells can be induced by an indirect bystander-type mechanism both in vitro and in vivo provides a potential link between these two untargeted effects and there are radiation responses in vivo consistent with the microenvironment contributing secondary cell damage as a consequence of an inflammatory-type response to radiation-induced injury. Intercellular signalling, production of cytokines and free radicals are features of inflammatory responses that have the potential for both bystander-mediated and persisting damage as well as for conferring a predisposition to malignancy. The induction of bystander effects and instabilities may reflect interrelated aspects of a non-specific inflammatory type response to radiation-induced stress and injury and be involved in a variety of the pathological consequences of radiation exposures. PMID- 12556328 TI - Radiation-induced genomic instability in immortalized haemopoietic stem cells. AB - PURPOSE: To investigate the expression of radiation-induced delayed reproductive death and chromosomal instability using an immortalized cell line (R-M26/2-1) with the characteristics of long-term repopulating haemopoietic stem cells established from the CBA mouse strain. MATERIALS AND METHODS: R-M26/2-1 cells were gamma-irradiated and maintained in culture for up to 41 population doublings. At intervals, measurements were made of cloning efficiency and cells were examined for cytogenetic aberrations at eight and 24 population doublings. The p53 status and p53 phosphorylation were investigated by Western analysis and immunocytochemistry and production of intracellular reactive oxygen species was investigated by use of the fluorescent probe DCFH-DA. RESULTS: The descendants of cells surviving 4Gy gamma-irradiation exhibited a reduced colony-forming efficiency and expressed chromosomal instability independent of p53 function and with no evidence of enhanced production of reactive oxygen species. The delayed reproductive death phenotype persisted at a constant rate of 12% clonogenic cell loss when colony formation was assessed in anchorage-dependent conditions on tissue culture substrates. However, R-M26/2-1 cells, like normal haemopoietic cells, can be cultured in anchorage-independent conditions and this type of assay demonstrated a 50% or greater persisting clonogenic cell loss. There was no significant delayed reproductive death or chromosomal instability in cultures established with 0.5 Gy gamma-irradiated R-M26/2-1 cells. CONCLUSIONS: A radiation-induced genomic instability phenotype, independent of p53 function and with no evidence of oxidative stress, was demonstrated in the descendants of 4Gy gamma-irradiated R-M26/2-1 cells and unstable aberrations characteristic of radiation-induced chromosomal instability may account for a component of the delayed reproductive death phenotype. Colony-forming efficiency and expression of the delayed death phenotype determined using an anchorage-independent assay was significantly greater than that determined using an anchorage-dependent assay indicating that some aspect of adherence influences these endpoints. The absence of significant instability in the descendants of 0.5 Gy gamma-irradiated cells implies a threshold for these endpoints in this haemopoietic stem cell line. PMID- 12556329 TI - Increased bystander mutagenic effect in DNA double-strand break repair-deficient mammalian cells. AB - PURPOSE: We have shown previously that when monolayer cultures of Chinese hamster ovary (CHO) cells are exposed to very low fluences of alpha-particles, HPRT mutations are induced in non-irradiated 'bystander' cells in the population. The present investigation was designed to examine the role of DNA repair in this process. MATERIALS AND METHODS: The DNA double-strand repair-deficient mutant cell line xrs-5 was exposed to mean doses of alpha-particles as low as 0.04 cGy whereby less than 1% of the nuclei were traversed by an alpha track and thus received any radiation exposure. RESULTS: With this very low alpha-particle fluence, most of the cells in the xrs-5 population appeared to be at risk for the induction of mutations, indicating a much larger bystander effect than observed with wild-type CHO cells. Molecular structural analyses showed that xrs-5 mutants primarily involved partial and total gene deletions as opposed to wild-type cells where point mutations predominated in bystander cells. CONCLUSIONS: These results indicate a very large bystander effect in xrs-5 cells. They support the hypothesis that unrepaired or misrepaired double-strand breaks (DSB), arising from opposed DNA lesions, enhance the sensitivity of bystander cells in xrs-5 cultures to the induction of mutations. PMID- 12556330 TI - Biophysical model of the radiation-induced bystander effect. AB - PURPOSE: To construct a quantitative model of the radiation-induced bystander effect based on diffusion-type spreading of bystander signal communication between the hit and non-hit cells. Cell inactivation and induced oncogenic transformation by broad- and microbeam irradiation systems are considered. MATERIALS AND METHODS: The biophysical model ByStander Diffusion Modelling (BSDM) postulates that the oncogenic bystander response observed in non-hit cells originates from specific signals received from inactivated cells. The bystander signals are assumed to be protein-like molecules spreading in the culture media by Brownian motion. The bystander signals are assumed to switch cells into a state of cell death (apoptotic/mitotic/necrosis) or induced oncogenic transformation modes. RESULTS: The bystander cell survival observed after treatment with the irradiated conditioned medium (ICM) using the broad-beam and the microbeam irradiation modalities were analysed and interpreted in the framework of the BSDM model. The model predictions for cell inactivation and induced oncogenic transformation frequencies agree well with observed data from micro and broad-beam experiments. In the case of irradiation with constant fraction of cells, transformation frequency for the bystander effect increases with increasing radiation dose. CONCLUSIONS: Bystander modelling based on diffusion of signals is in good agreement with experimental cell survival data and induced oncogenic transformation frequencies. The data confirm the protein like nature of the bystander signal. Linear extrapolation of the cell response to low doses of radiation might underestimate carcinogenic risk, for example for domestic radon hazards, if the contribution from the bystander effect is neglected. The BSDM predicts that the bystander effect cannot be interpreted solely as a low-dose effect phenomenon. It is shown that the bystander component of radiation response can increase with dose and be observed at high doses as well as at low doses. The validity of this conclusion is supported by analysis of experimental results from high-linear energy transfer microbeam experiments. PMID- 12556331 TI - Expression of the DNA-PK binding protein E4-34K fails to confer radiation sensitivity to mammalian cells. AB - PURPOSE: The adenovirus E4orf6 34 kDa protein (E4-34k) is known to disrupt V(D)J recombination as a result of its interaction with the catalytic subunit of cellular DNA-dependent protein kinase (DNA-PK(cs)), a major participant in the repair of DNA double-strand breaks (DSB). Previous studies have shown that cells with disrupted DSB repair and V(D)J recombination due to attenuation of DNA PK(cs) activity exhibit a radiation-sensitive phenotype. It is not known at present whether the E4-34k protein can also modify cellular response to ionizing radiation. In an attempt to develop a novel gene therapy strategy to modify cellular radiation response, we sought to determine if expression of the adenovirus E4-34k protein resulted in sensitization to clinically relevant doses of ionizing radiation. MATERIALS AND METHODS: In order to minimize potential bias resulting from selection procedures, we performed clonogenic survival assays on DU 145 prostate cancer cells, RKO colorectal cancer cells and 293 kidney cells following transient transfection of E4-34k- and/or E1B-55k-expressing plasmids. Western blots and immunohistochemical analyses were used to demonstrate E4-34k expression within transfected cells. FACS sorting was carried out to enrich cells transfected with a plasmid that expresses both E4-34k and enhanced green fluorescent protein. RESULTS: It is shown that E4-34k expression does not affect cellular radiosensitivity of transiently transfected populations of either DU 145 prostate or RKO colon cancer cell lines. Similarly, the radiosensitivity of human embryonic kidney 293 cells, which constitutively express the E1B-55k protein, was also unaffected. The radiosensitivity of DU 145 cells co-transfected with E4-34k- and E1-55K-expressing plasmids was unchanged, suggesting that the adenovirus E1B 55k protein does not augment any effects E4-34k might have on DNA-PK(cs) activity. CONCLUSIONS: The lack of radiosensitization by E4-34k expression is quite intriguing as it is known that E4-34k interaction with DNA-PK(cs) causes disruption of V(D)J recombination, a process dependent on DSB rejoining. These data suggest that for future studies, preferential targeting of DNA-PK(cs) DSB activity will be required to influence cellular radiosensitivity. PMID- 12556332 TI - Role of DNA-PK in the process of aberration formation as studied in irradiated human glioblastoma cell lines M059K and M059J. AB - PURPOSE: The participation of various DNA double-strand break repair mechanisms in the formation of chromosome aberrations is not yet fully understood. To investigate particularly the role of non-homologous end-joining, we analysed the formation of radiation-induced aberrations in a DNA-protein kinase (PK(CS)) proficient cell line M059K and in a deficient cell line M059J. MATERIALS AND METHODS: Plateau-phase M059K and M059J cells were irradiated with low doses of X rays. Chromosome aberrations were determined as genomic yields of dicentric chromosomes and excess acentric fragments, scored in Giemsa-stained metaphases, and as partial yields of reciprocal translocations and total visible complex exchanges (complex aberrations) for chromosomes 4, 7 and 11 using the FISH method. M059K cells were also analysed in the presence of 50 micro m wortmannin, a DNA-PK inhibitor. RESULTS: DNA-PK-deficient cells showed a higher yield of simple stable and unstable and of complex aberrations in comparison with DNA-PK proficient cells. The largest differences were observed for excess acentric fragments and for complex aberrations. DNA-PK inhibition by wortmannin in M059-K cells resulted in increased aberration yield in the same qualitative and quantitative manner as in M059-J cells. CONCLUSIONS: The results obtained with DNA-PK-deficient M059J cells and with DNA-PK-proficient M059K cells treated with wortmannin, an inhibitor of DNA-PK and ATM, suggest that the elimination of DNA PK-dependent non-homologous end-joining can recruit a slow, error-prone repair process, which is DNA-PK independent and favours the increased formation of chromosome aberrations. The nature of this pathway and the way of its participation in aberration formation need further elucidation. PMID- 12556334 TI - Routine screening mammography: how important is the radiation-risk side of the benefit-risk equation? AB - The potential radiation hazards associated with routine screening mammography, in terms of breast cancer induction, are discussed in the context of the potential benefits. The very low energy X-rays used in screening mammography (26-30 kVp) are expected to be more hazardous, per unit dose, than high-energy X- or gamma rays, such as those to which A-bomb survivors (from which radiation risk estimates are derived) were exposed. Based on in vitro studies using oncogenic transformation and chromosome aberration end-points, as well as theoretical estimates, it seems likely that low doses of low-energy X-rays produce an increased risk per unit dose (compared with high energy photons) of about a factor of 2. Because of the low doses involved in screening mammography, the benefit-risk ratio for older women would still be expected to be large, though for younger women the increase in the estimated radiation risk suggests a somewhat later age than currently recommended--by about 5-10 years--at which to commence routine breast screening. PMID- 12556335 TI - Response of mammalian cell lines that undergo necrosis after ionizing irradiation to DNA-incorporated tritium and iodine-125 decays: implications for models of radiation action. AB - PURPOSE: To test the validity of enzymatic DNA repair-based models of ionizing radiation-induced mammalian cell killing. MATERIALS AND METHODS: Sensitivity to ionizing radiation-induced killing was determined by clonogenic survival assay. RESULTS: A panel of seven cell lines that have apparently normal enzymatic DNA repair capability and die predominantly by necrosis were shown to have marked differences in their sensitivity to killing by gamma- or X-irradiation. Despite such differences, the pseudo-diploid cell lines CHO, V79, and Vero (when corrected for differences in cell cycle distribution) and normal mouse embryo fibroblasts all had comparable D(o) values for killing by DNA-incorporated (125)I that were in the range of 57-66 decays; whilst the near-tetraploid or-triploid B16-F1, RUC-2 and SQ-20B cell lines had D(o) values of around 130 (125)I decays. The sensitivity of these cell lines to killing by DNA-incorporated (3)H decays was similarly related to ploidy. CONCLUSIONS: Differences in sensitivity to killing by DNA-incorporated (3)H or (125)I decays between the cell lines tested were primarily related to differences in ploidy and, second, to differences in cell cycle distribution or nuclear volume. The data do not support suggestions that generalized variability in an hypothetical aspect of enzymatic DNA repair can explain differences in sensitivity to killing by external-beam, low-linear energy transfer ionizing radiation. PMID- 12556336 TI - Transcription-based model for the induction of interchromosomal exchange events by ionizing irradiation in mammalian cell lines that undergo necrosis. AB - PURPOSE: To develop a model of the molecular events leading to ionizing radiation induced interchromosomal exchange in mammalian cells that undergo necrosis. OUTLINE OF THE MODEL: DNA double-strand breaks (DSB), primarily those involving multiple individual damage sites (i.e. complex DSB), are postulated to be the critical initiating lesion. Only those DSB occurring in transcription units that are associated with transcription factories are postulated to induce chromosomal exchange events. It is suggested that such DSB are brought into contact with a DNA topoisomerase I (topo I) molecule through RNA polymerase II (pol II) catalysed transcription and give rise to trapped DNA-topo I cleavage complexes. Trapped complexes are postulated to interact with another topo I molecule on a temporarily inactive transcription unit at the same transcription factory leading to DNA cleavage and subsequent strand exchange between the cleavage complexes. Interchromosomal exchange events will occur when the cleavage complexes are on transcription units located on different chromosomes. OUTCOMES: The model can explain a variety of phenomena that includes: the increased resistance to killing by ionizing radiation shown by quiescent cells (i.e. potentially lethal damage repair); the effect of ploidy on sensitivity to killing by DSB; the effect of chasing time on sensitivity to killing by DNA-incorporated (125)I decays after pulse labelling; and the size of the target for ionizing radiation-induced cell killing. PMID- 12556337 TI - Comparison of repair and rejoining fidelity between two isogenic human ovarian carcinoma cell lines. AB - PURPOSE: The difference in radiosensitivity between two isogenic tumour cell lines was evaluated to determine whether factors such as sublethal and potentially damage repair, DNA double-strand break repair and fidelity of repair can be related to differences in radiosensitivity. MATERIALS AND METHODS: The cell lines used were the ovarian carcinoma A2780s and a radiation-resistant derivative A2780cp. Radiation response was measured in terms of cell survival, recovery of sublethal (SLD) and potentially lethal damage (PLD), induction of and recovery of DNA strand breaks, and fidelity of DNA repair using a cell-free plasmid assay. RESULTS: While A2780cp was more resistant to radiation than A2780s, it showed less ability for recovery of SLD and PLD. DNA strand-break induction was the same for both cell lines, while only at very high doses did A2780cp show greater DNA strand-break recovery than A2780s. Fidelity of rejoining DNA was significantly higher in the A2780cp cell line. CONCLUSION: The difference in radiosensitivity between these two cell lines was not related to recovery of PLD or SLD or to the induction of DNA damage. It appears that fidelity of DNA rejoining, which was significantly higher in the resistant cell line, may be related to the difference in radiosensitivity. PMID- 12556338 TI - Computer analysis of mFISH chromosome aberration data uncovers an excess of very complicated metaphases. AB - PURPOSE: To analyse spectra of chromosome aberrations induced in vitro by low LET radiation, in order to characterize radiation damage mechanisms quantitatively. METHODS: Multiplex fluorescence in situ hybridization (mFISH) allows the simultaneous identification of each homologous chromosome pair by its own colour. mFISH data, specifying number distributions for colour junctions in metaphases of human peripheral blood lymphocytes 72 hours after exposure in vitro to a 3 Gy gamma-ray dose, were combined with similar, previously published results. Monte Carlo computer implementations of radiobiological models for chromosome aberration production guided quantitative analyses, which took into account distribution of cells among different metaphases and lethal effects or preferential elimination of some aberrations at cell division. RESULTS AND CONCLUSIONS: Standard models of DNA damage induction/repair/misrepair explain the main trends of the data as regards the fraction of metaphases having a particular number of colours involved in colour junctions. However, all standard models systematically under-predict the observed fraction of metaphases where a large number of different chromosomes participate in aberrations. An early appearance of chromosomal instability could explain most of the discrepancies. PMID- 12556339 TI - Chromosomal radiosensitivity study of temporary nuclear workers and the support of the adaptive response induced by occupational exposure. AB - PURPOSE: To study chromosomal radiosensitivity in a population of radiation workers and investigate the possibility of an adaptive response in lymphocytes of workers after short-term occupational exposure to ionizing radiation. MATERIALS AND METHODS: The studied group comprised 41 workers temporarily employed at the Nuclear Power Plant Doel (Belgium) for reactor maintenance. A blood sample was taken before and directly after the exposure period of about 1 month. Chromosomal radiosensitivity was assessed in vitro by the G2 assay and the G0 micronucleus (MN) assay. For the MN assay, a low dose-rate (LDR) in vitro irradiation protocol was applied in addition to high dose-rate (HDR) irradiation of the blood samples in order to determine the dose-rate sparing (DRS) effect. RESULTS: No statistically significant effect of the occupational exposures (up to 10 mSv) on the baseline MN frequencies without in vitro irradiation was observed. A comparison of the number of chromatid aberrations pre- and post-exposure shows no effect of the occupational exposure. On the other hand, the G0-MN assay with the LDR irradiation protocol reveals a systematic reduction in chromosomal radiosensitivity by the exposure, increasing with dose. For workers who received the highest dose (4-10 mSv) a statistically significant (p <0.05) decrease of the in vitro induced MN yields and increase of the dose-rate sparing was observed. CONCLUSIONS: Short-term low-dose occupational exposure may act as an in vivo adaptive dose and stimulate repair in G0 lymphocytes. PMID- 12556340 TI - Alterations in water and electrolyte absorption in the rat colon following neutron irradiation: influence of neutron component and irradiation dose. AB - PURPOSE: To study the absorptive function of rat colon following whole-body exposure to neutron irradiation, either to the same total dose with varying proportion of neutrons or to the same neutron proportion with an increasing irradiation dose. MATERIALS AND METHODS: Different proportions of neutron irradiation were produced from the reactor SILENE using a fissile solution of uranium nitrate (8, 47 and 87% neutron). Water and electrolyte fluxes were measured in the rat in vivo under anaesthesia by insertion into the descending colon of an agarose gel cylinder simulating the faeces. Functional studies were completed by histological analyses. In the first set of experiments, rats received 3.8 Gy with various neutron percentages and were studied from 1 to 14 days after exposure. In the second set of experiments, rats were exposed to increasing doses of irradiation (1-4Gy) with a high neutron percentage (87%n) and were studied at 4 days after exposure. RESULTS AND CONCLUSIONS: The absorptive capacity of rat colon was diminished by irradiation at 3-5 days, with a nadir at 4 days. The results demonstrate that an increase in the neutron proportion is associated with an amplification of the effects. Furthermore, a delay in the re establishment of normal absorption was observed with the high neutron proportion (87%n). A dose-dependent reduction of water absorption by rat colon was also observed following neutron irradiation (87%n), with a 50% reduction at 3 Gy. Comparison of this dose-effect curve with the curve obtained following gamma (60)Co-irradiation indicates an RBE of 2.2 for absorptive colonic function in rat calculated at 4 days after exposure. PMID- 12556341 TI - Evidence for the involvement of DNA-dependent protein kinase in the phenomena of low dose hyper-radiosensitivity and increased radioresistance. AB - PURPOSE: To investigate the role of DNA-dependent protein kinase (DNA-PK) in the phenomena of low dose hyper-radiosensitivity (HRS) and increased radioresistance (IRR) using the genetically related M059 cell lines of disparate PRKDC status. MATERIALS AND METHODS: Clonogenic survival was measured for the three cell lines following low doses of X-irradiation using a flowactivated cell sorting (FACS) plating technique. The presence of PRKDC, G22p1 and Xrcc5 proteins was determined by Western blotting and a kinase assay used to measure DNA-PK complex activity. RESULTS: The survival responses for the three cell lines over the 0-0.3Gy dose range were comparable, but differences in radiosensitivity were evident at doses >0.4Gy. M059K and M059J/Fus1 cells (both PRKDC competent) exhibited marked HRS/IRR responses, albeit to different extents. M059J cells (PRKDC incompetent) were extremely radiosensitive exhibiting a linear survival curve with no evidence of IRR. The presence of IRR was coincident with the presence of PRKDC protein and functional DNA-PK activity. CONCLUSIONS: HRS is a response that is independent of DNA-PK activity. In contrast, IRR showed a dependence on the presence of PRKDC protein and functional DNA-PK activity. These data support a role for DNA-PK activity in the IRR response. PMID- 12556342 TI - The radiation hypersensitivity of cells at mitosis. AB - PURPOSE: Mitotic cells are hypersensitive to ionizing radiation, exhibiting single-hit inactivation coefficients near to those of repair deficient cell lines and lymphocytes. To elucidate possible mechanisms for this hypersensitivity, the kinetics of oxygen radiosensitization, the proportion of indirect effect by OH radicals and the kinetics of radiation-induced DNA strand breakage in the chromatin of mitotic cells were investigated. MATERIALS AND METHODS: Synchronized populations of >90% mitotic HT-29 cells were obtained by the mitotic shake-off method. Cells were irradiated at < or =4 degrees C with (137)Cs gamma-rays. Cellular oxygen concentration was varied by gassing cell suspensions prior to and during irradiation with mixtures of pure N(2) that contained 5% CO(2) and measured quantities of O(2). The indirect effect of OH radicals was investigated with the radical scavenger, DMSO. DNA strand breakage was measured by the comet assay. RESULTS: Mitotic HT-29 cell inactivation is well described by a single-hit inactivation coefficient (alpha) of 1.14 +/- 0.06 Gy(-1). The oxygen enhancement ratio of mitotic cells (at 10% survival) was found to be approximately 2.0, significantly lower than the value of 2.8 measured for interphase (asynchronous) cells. More than 60% of mitotic cell killing was eliminated when the media contained 2 M DMSO, indicating that indirect effect is as important in the killing of mitotic cells as it is for interphase cells. The chromatin in mitotic cells was found to be ~2.8 times more sensitive to radiation-induced DNA single strand breakage than the chromatin of interphase cells. CONCLUSIONS: The alpha inactivation coefficient of mitotic HT-29 cells was ~30 times larger than that of interphase cells. Mitotic cell chromatin appears to contain intrinsic DNA breaks that are not lethal. In addition, chromatin in mitotic cells was found to be more susceptible to radiation-induced DNA strand-breakage than the dispersed chromatin of interphase cells. How the enhanced production of these simple DNA lesions (that are usually reparable) translates into the lethal (non-reparable) events associated with alpha-inactivation is not known. The compaction/dispersion status of DNA throughout the cell cycle appears to be an important factor for determining intrinsic cell radiosensitivity and might be manipulated for radiotherapeutic advantage. PMID- 12556343 TI - Explanation of protective effects of low doses of gamma-radiation with a mechanistic radiobiological model. AB - PURPOSE: To test whether data that show protective effects of low doses against spontaneous neoplastic transformation of C3H 10T1/2 cells can be explained with a biomathematical model that includes radioprotective mechanisms. To link important features of the model to known biological processes. MATERIALS AND METHODS: The model simulates double-strand break formation in transcriptionally active and in bulk DNA, translocation of DNA segments, and the fixation of damage at mitosis; promotion is also included. The model equations were solved numerically using a stiff solver. RESULTS: The data were successfully simulated by the model: cell transformation-reducing effects of low doses of gamma-radiation delivered at low dose-rates are explained by radiation-inducible DNA repair and enzymatic scavenging. CONCLUSIONS: The model successfully simulates experimental data. The highly nonlinear features of the data point to a nonlinear dose-effect relationship at low doses and indicate that linear extrapolation from moderate (or high) to low doses and dose-rates may not be justified for in vitro studies of the cell line under consideration. PMID- 12556344 TI - Radiation-induced up-regulation of telomerase in KG1a cells is influenced by dose rate and radiation quality. AB - PURPOSE: To examine the influence of dose, dose-rate and radiation quality on telomerase activity (TA) in the KG1a hematopoietic cell line. MATERIALS AND METHODS: KG1a cells were irradiated with gamma-rays (0.5-5 Gy) at 0.025 Gy/min, 0.30 Gy/min and 1.57 Gy/min and with a neutron/gamma-ray field (5 Gy). Cell viability was determined by trypan blue exclusion. Apoptosis and cell cycle distribution were evaluated by flow cytometry. Proliferative capacity was studied by MTS assay and TA by PCR. Following 3Gy gamma-irradiation, the expression of hTERT, hTR and TP1 genes was evaluated by RT-PCR. RESULTS: Dose- and dose-rate dependent telomerase activation with an increase in hTERT mRNA and a drop in hTP1 mRNA were observed after irradiation. Down-regulation of telomerase activity occurred in a dose-dependent manner. Although non-significant changes in short term survival were observed after irradiation, late apoptosis became evident after G2/M arrest. Early repression of TA preceded telomerase activation in samples irradiated with a neutron/gamma-ray field, in which short-term survival was affected. CONCLUSIONS: Radiation-induced telomerase activation depends on dose-rate. High-LET and low-LET irradiations induce similar changes in TA that differ mainly in their kinetics and their magnitude. Changes in TA are not related to cell-cycle redistribution nor to the induction of cell death; they are the consequence of specific regulatory responses to ionizing radiation. Mechanisms including both transcriptional and post-translational control may be involved in this regulation. PMID- 12556345 TI - Increased antioxidant potential of combined topical vitamin E and C against lipid peroxidation of eicosapentaenoic acid in pig skin induced by simulated solar radiation. AB - PURPOSE: To study the protective effect of topically applied vitamin E (TOC), vitamin C (ASC), or a combination of both, against the lipid peroxidation of eicosapentaenoic acid (EPA) induced by simulated solar radiation (SSR). MATERIAL AND METHODS: EPA (25 nmol cm(-2)) was topically applied to pig skin explants, followed by increasing doses of TOC and ASC, either alone or combined. Epidermal lipid peroxidation was assessed after 15 min of exposure to SSR (resulting in a UVA and UVB dose of 18 and 3 kJ m(-2), respectively). SSR-induced changes in the levels of TOC and ASC were determined in the stratum cornaeum and the viable epidermis. RESULTS: SSR exposure of EPA-treated pig skin resulted in a twofold increase in epidermal lipid peroxidation (p <0.005) which was reduced by topically applied TOC or ASC 60 min before SSR exposure (p <0.05). Compared with TOC (5 nmol cm(-2)), a 400-fold higher ASC dose was needed and only TOC provided complete protection against the lipid peroxidation of EPA. The levels of both TOC and ASC clearly increased in both skin compartments by increasing the applied dose of these two compounds (p <0.05). In contrast to ASC, TOC was consumed by up to 55-70% during SSR exposure (p <0.05). Compared with separate application, combined TOC and ASC efficiently protected against lipid peroxidation of EPA at doses that were five and 200 times lower, respectively. In the presence of low ASC doses, 70-100% of epidermal TOC was regenerated during SSR exposure (p <0.05). CONCLUSIONS: Topically applied TOC and ASC protect against SSR-induced lipid peroxidation of EPA. The synergism between TOC and ASC resulted in a more efficient protection at substantially lower doses of both antioxidants. Co supplementation of EPA with TOC and/or ASC might improve the beneficial biological effects of EPA. PMID- 12556346 TI - A forty-year memoir of research on the regulation of glucose transport into muscle. AB - This historical review describes the research on the regulation of glucose transport in skeletal muscle conducted in my laboratory and in collaboration with a number of colleagues in other laboratories. This research includes studies of stimulation of glucose transport, GLUT4 translocation, and GLUT4 expression by exercise/muscle contractions, the role of Ca(2+) in these processes, and the interactions between the effects of exercise and insulin. Among the last are the additive effects of insulin and contractions on glucose transport and GLUT4 translocation and the increases in muscle insulin sensitivity and responsiveness induced by exercise. PMID- 12556347 TI - The melanocortin system. AB - The melanocortin system consists of melanocortin peptides derived from the proopiomelanocortin gene, five melanocortin receptors, two endogenous antagonists, and two ancillary proteins. This review provides an abbreviated account of the basic biochemistry, pharmacology, and physiology of the melanocortin system and highlights progress made in four areas. In particular, recent pharmacological and genetic studies have affirmed the role of melanocortins in pigmentation, inflammation, energy homeostasis, and sexual function. Development of selective agonists and antagonists is expected to further facilitate the investigation of these complex physiological functions and provide an experimental basis for new pharmacotherapies. PMID- 12556348 TI - Postnatal oxytocin alleviates adverse effects in adult rat offspring caused by maternal malnutrition. AB - Repeated oxytocin administration to adult rats causes a long-term decrease of plasma levels of corticosterone and blood pressure and stimulates growth and fat retention. Maternal undernutrition increases blood pressure and plasma corticosterone in adult offspring. We hypothesized that oxytocin treatment early in life would alleviate adverse effects of intrauterine food restriction. Male pups from ad libitum-fed and food-restricted (fed 60% of ad libitum intake) dams were injected with oxytocin or saline in days 1-14 after birth. At 4 mo, blood pressure, plasma levels of corticosterone, and adiposity were assessed. Oxytocin treatment decreased blood pressure independently of nutrition, whereas the increased plasma levels of corticosterone were lowered to normal levels in food restricted offspring. Blood pressure and adiposity were not affected by in utero food restriction, whereas birth and adult weight were. In conclusion, postnatal events may alleviate adverse effects caused by in utero food restriction. In contrast to more severe food restriction, a moderate general food restriction during gestation had no effect on blood pressure in the offspring. PMID- 12556349 TI - Reduced amino acid availability inhibits muscle protein synthesis and decreases activity of initiation factor eIF2B. AB - We have examined the effect of a hemodialysis-induced 40% reduction in plasma amino acid concentrations on rates of muscle protein synthesis and breakdown in normal swine. Muscle protein kinetics were measured by tracer methodology using [(2)H(5)]phenylalanine and [1-(13)C]leucine and analysis of femoral arterial and venous samples and tissue biopsies. Net amino acid release by muscle was accelerated during dialysis. Phenylalanine utilization for muscle protein synthesis was reduced from the basal value of 45 +/- 8 to 25 +/- 6 nmol x min(-1) x 100 ml leg(-1) between 30 and 60 min after start of dialysis and was stimulated when amino acids were replaced while dialysis continued. Muscle protein breakdown was unchanged. The signal for changes in synthesis appeared to be changes in plasma amino acid concentrations, as intramuscular concentrations remained constant throughout. The changes in muscle protein synthesis were accompanied by a reduction or stimulation, respectively, in the guanine nucleotide exchange activity of eukaryotic initiation factor (eIF)2B following hypoaminoacidemia vs. amino acid replacement. We conclude that a reduction in plasma amino acid concentrations below the normal basal value signals an inhibition of muscle protein synthesis and that corresponding changes in eIF2B activity suggest a possible role in mediating the response. PMID- 12556350 TI - PPARalpha /gamma ragaglitazar eliminates fatty liver and enhances insulin action in fat-fed rats in the absence of hepatomegaly. AB - Peroxisome proliferator-activated receptor (PPAR)alpha and PPARgamma agonists lower lipid accumulation in muscle and liver by different mechanisms. We investigated whether benefits could be achieved on insulin sensitivity and lipid metabolism by the dual PPARalpha/gamma agonist ragaglitazar in high fat-fed rats. Ragaglitazar completely eliminated high-fat feeding-induced liver triglyceride accumulation and visceral adiposity, like the PPARalpha agonist Wy-14643 but without causing hepatomegaly. In contrast, the PPARgamma agonist rosiglitazone only slightly lessened liver triglyceride without affecting visceral adiposity. Compared with rosiglitazone or Wy-14643, ragaglitazar showed a much greater effect (79%, P < 0.05) to enhance insulin's suppression of hepatic glucose output. Whereas all three PPAR agonists lowered plasma triglyceride levels and lessened muscle long-chain acyl-CoAs, ragaglitazar and rosiglitazone had greater insulin-sensitizing action in muscle than Wy-14643, associated with a threefold increase in plasma adiponectin levels. There was a significant correlation of lipid content and insulin action in liver and particularly muscle with adiponectin levels (P < 0.01). We conclude that the PPARalpha/gamma agonist ragaglitazar has a therapeutic potential for insulin-resistant states as a PPARgamma ligand, with possible involvement of adiponectin. Additionally, it can counteract fatty liver, hepatic insulin resistance, and visceral adiposity generally associated with PPARalpha activation, but without hepatomegaly. PMID- 12556351 TI - Fiber type-specific determinants of Vmax for insulin-stimulated muscle glucose uptake in vivo. AB - The aim of this study was to determine barriers limiting muscle glucose uptake (MGU) during increased glucose flux created by raising blood glucose in the presence of fixed insulin. The determinants of the maximal velocity (V(max)) of MGU in muscles of different fiber types were defined. Conscious rats were studied during a 4 mU x kg(-1) x min(-1) insulin clamp with plasma glucose at 2.5, 5.5, and 8.5 mM. [U-(14)C]mannitol and 3-O-methyl-[(3)H]glucose ([(3)H]MG) were infused to steady-state levels (t = -180 to 0 min). These isotope infusions were continued from 0 to 40 min with the addition of a 2-deoxy-[(3)H]glucose ([(3)H]DG) infusion. Muscles were excised at t = 40 min. Glucose metabolic index (R(g)) was calculated from muscle-phosphorylated [(3)H]DG. [U-(14)C]mannitol was used to determine extracellular (EC) H(2)O. Glucose at the outer ([G](om)) and inner ([G](im)) sarcolemmal surfaces was determined by the ratio of [(3)H]MG in intracellular to EC H(2)O and muscle glucose. R(g) was comparable at the two higher glucose concentrations, suggesting that rates of uptake near V(max) were reached. In summary, by defining the relationship of arterial glucose to [G](om) and [G](im) in the presence of fixed hyperinsulinemia, it is concluded that 1) V(max) for MGU is limited by extracellular and intracellular barriers in type I fibers, as the sarcolemma is freely permeable to glucose; 2) V(max) is limited in muscles with predominantly type IIb fibers by extracellular resistance and transport resistance; and 3) limits to R(g) are determined by resistance at multiple steps and are better defined by distributed control rather than by a single rate-limiting step. PMID- 12556352 TI - Improved beta-cell function after standardized weight reduction in severely obese subjects. AB - Islet function was examined in 13 severely obese women [body mass index 46.4 +/- 5.5 (SD) kg/m(2)] before and after standardized 15 and 25% weight reduction (WR) instituted by bariatric surgery. The insulin response to arginine at fasting (AIR(1)), at 14 mmol/l, and at >25 mmol/l glucose was reduced by 37-50% after 15 and 25% WR (P /=20). Genes with fold change values of four or more were matched with loci linked to glaucoma, by using gene databases. RESULTS: TIGR/MYOC induction by DEX occurred only in HTM cells. Secretory and glycosylation characteristics remained the same across cell types. Expression profile analysis revealed multiple genes differentially upregulated in HTM-DEX including, in addition to TIGR/MYOC, a serine protease inhibitor (alpha1 antichymotrypsin), a neuroprotective factor (pigment epithelium-derived factor), an antiangiogenesis factor (cornea-derived transcript 6), and a prostaglandin synthase (prostaglandin D(2) synthase). Fifteen of the 249 genes with fold change values of four or more mapped to glaucoma-linked loci. CONCLUSIONS: The induction of TIGR/MYOC by DEX is HTM-specific, whereas its secretory and glycosylation characteristics are ubiquitous. The known functions of HTM-DEX-specific genes reveal the presence of protective and damaging mechanisms for regulation of IOP during DEX treatment. Besides TIGR/MYOC, other HTM-DEX-specific genes may be good candidates for linkage to glaucoma. PMID- 12556372 TI - Susceptibility to retinal light damage in transgenic rats with rhodopsin mutations. AB - PURPOSE: To determine relative light-induced retinal damage susceptibility in transgenic rats expressing mutations in the N- or C-terminal region of rhodopsin. METHODS: Heterozygous transgenic rats, including P23H sublines 2 and 3 and S334ter sublines 4 and 9, were reared in dim cyclic light or in darkness before visible light exposure starting at various times of the day or night. Before exposure to light, some rats were given the synthetic antioxidant dimethylthiourea (DMTU). At various times after intense light treatment, rats were killed for determinations of rhodopsin and retinal DNA recovery, DNA fragmentation patterns, and Northern blot analysis of retinal heme oxygenase (HO) 1 and interphotoreceptor retinol binding protein (IRBP). Rod outer segments (ROSs) were isolated for Western blot analysis of rhodopsin using N- and C- terminal-specific monoclonal antibodies. RESULTS: All rats incurred greater photoreceptor cell damage from exposure to light starting at 1 AM than from exposure at 5 PM. Among cyclic-light-reared rats, P23H line 3 animals were more susceptible to light-induced damage than P23H line 2 animals. S334ter rats exhibited retinal light damage profiles similar to those in normal rats. Dark rearing potentiated retinal damage by light. However, dark-rearing alone prolonged photoreceptor cell life in P23H rats, but had no such effect in S334ter animals. DMTU pretreatment was effective in preventing or reducing light-induced retinal damage in all transgenic rats. S334ter rat ROSs contained the truncated form of rhodopsin. Intense light exposure resulted in DNA ladders typical of apoptotic cell death and the simultaneous induction of retinal HO-1 mRNA and reduced expression of IRBP. CONCLUSIONS: Light-induced retinal damage in transgenic rats depends on the time of day of exposure to light, prior light-or dark-rearing environment, and the relative level of transgene expression. Retinal light damage leads to apoptotic visual cell loss and appears to result from oxidative stress. These results suggest that reduced environmental lighting and/or antioxidant treatment may delay retinal degenerations arising from rhodopsin mutations. PMID- 12556373 TI - Cross-sectional study of visual acuity and electroretinogram in two types of dominant drusen. AB - PURPOSE: To compare the changes with increasing age of ERG parameters in relation to clinical data in two distinct phenotypes of genetically determined, dominantly inherited macular drusen: malattia leventinese (ML) and Zermatt macular dystrophy (ZMD). METHODS: Ganzfeld rod- and cone-electroretinograms (ERGs) from 15 patients affected with ML and 14 patients with ZMD and clinical data were analyzed retrospectively. The patients' ages ranged from 20 to 77 years in the ML group and from 9 to 74 years in the ZMD group. RESULTS: Both inherited macular degenerations caused a marked decrease in visual acuity, the latest after age 65. Most patients with ML retained good visual function (0.8-1.0) until the fifth decade, followed by a rapid decrease in the fifth or sixth decade. ZMD is characterized by a relatively continuous decrease in visual acuity with increasing age. Morphologically, in the juvenile stages in both entities, drusen were observed at the posterior pole. Rod-driven and cone-driven ERG b-wave amplitudes decreased nearly linearly in ML and ZMD in accord with the normal loss of amplitude with increasing age. Implicit times of cone b-waves for ML increased markedly with age, whereas in ZMD the values were always prolonged beyond the normal range with a slight increase with age. CONCLUSIONS: In terms of visual acuity, the progression of both dominantly inherited macular dystrophies is quite different. This is not reflected in the amplitudes of the b-waves in the Ganzfeld ERGs, which decrease normally for both entities. Implicit times of the cone-b waves were more markedly prolonged in ML compared with ZMD. In-depth longitudinal documentation of the natural course of those dominantly inherited macular diseases should facilitate patient counseling. PMID- 12556374 TI - Identification of infants with significant refractive error and strabismus in a population screening program using noncycloplegic videorefraction and orthoptic examination. AB - PURPOSE: The second Cambridge Infant Vision Screening Program examined whether screening for accommodative errors by using videorefraction without cycloplegia could effectively serve as a first stage of screening for refractive errors, measured by standard cycloplegic retinoscopy. The screening also included an orthoptic examination for detection of strabismus. METHODS: All infants born in the Cambridge (UK) Health District, over a 2-year period, were invited for screening. Of those 5142 (76%) with mean age 8.1 +/- 0.8 months (SD) attended and received noncycloplegic videorefraction and an orthoptic examination. All those with a focusing error or orthoptic problem, as well as a randomly selected sample of visually normal control subjects, were invited to follow-up a month later for cycloplegic retinoscopy, repeat noncycloplegic videorefraction and orthoptic examination. RESULTS: Of the 5142 screened, 514 had a focusing error or orthoptic problem (positives). Four hundred thirty-nine of these and 284 visually normal control subjects (negatives) attended follow-up. A refractive or orthoptic condition was confirmed in 59.0% of the positive cases, whereas infants in 96.8% of the negative cases were confirmed normal. Adjusting for the proportions of the population represented by those infants seen at follow-up, sensitivity for the screening procedure was calculated at 0.67 and specificity at 0.96. Detailed results are presented in terms of the different conditions detected at screening (far, near, and anisometropic focus and orthoptic error), distribution of greatest axes at screening, and a comparison of initial videorefraction with repeat videorefraction and cycloplegic retinoscopy. CONCLUSIONS: A noncycloplegic screening procedure, simpler to perform than cycloplegic screening, succeeded in detecting a large proportion of infants with significant ametropia, particularly those with significant hyperopia, which is considered to be a strabismogenic and amblyogenic risk factor. PMID- 12556375 TI - Intraocular ciprofloxacin levels after oral administration in silicone oil-filled eyes. AB - PURPOSE: To evaluate penetration of oral ciprofloxacin in the retro-silicone oil space fluid (RSOF) in silicone oil (SO)-filled eyes. METHODS: One dose of 750 mg ciprofloxacin was given to two groups of five patients with vitrectomized eyes with SO endotamponade, 4 hours (group I) and 8 hours (group II) before SO removal. In 10 vitrectomized eyes with SO endotamponade (group III) and another 10 patients scheduled for vitrectomy for the first time (group IV), two 750-mg doses every 12 hours, with the last dose 12 hours before surgery, were given. Blood samples were taken at the time of collection of RSOF samples in groups I, II, and III and of the vitreous in group IV. All samples were assayed for ciprofloxacin by high-performance liquid chromatography. RESULTS: The mean drug concentration in the RSOF was 0.34 +/- 0.09, 0.37 +/- 0.04, 0.84 +/- 0.29, and 0.44 +/- 0.11 micro g/mL in groups I, II, III, and IV respectively. The mean serum concentration was 1.29 +/- 0.63, 1.08 +/- 0.14, 1.93 +/- 0.84, and 1.34 +/- 0.55 micro g/mL in groups I, II, III, and IV respectively with no statistically significant difference between groups III and IV (P = 0.081). CONCLUSIONS: Antibiotic levels in the RSOF in SO-filled eyes after oral administration of ciprofloxacin in two 750-mg doses exceeded the minimal inhibitory concentration for 90% of isolates (MIC(90)) for most bacterial species and was higher than levels reached in the vitreous in nonvitrectomized eyes (P = 0.001). PMID- 12556376 TI - Alcohol debridement of the corneal epithelium in PRK and LASEK: an electron microscopic study. AB - PURPOSE: To determine the plane of cleavage of the corneal epithelium and smoothness of underlying stroma, after alcohol debridement in photorefractive keratectomy (PRK) and laser subepithelial keratectomy (LASEK). METHODS: The epithelial flap from six patients undergoing alcohol delamination of corneal epithelium before PRK and the epithelium and stroma from three eye bank donor eyes were fixed and processed for transmission (TEM) and scanning electron microscopy (SEM). The smoothness of the underlying stroma was studied by SEM and the plane of cleavage was determined by morphologic examination and morphometric measurements of basement membrane attached to the epithelial flap, using image analysis software. RESULTS: A very smooth stromal bed, ideal for PRK was seen in the stroma of all three eye bank donor eyes after alcohol delamination. The plane of cleavage was determined to be at the hemidesmosomal attachments, including the most superficial part of the lamina lucida of the basement membrane. CONCLUSIONS: Alcohol delamination of the corneal epithelium before PRK or LASEK consistently results in a very smooth cleavage at the level of the hemidesmosomal attachments, including the superficial lamina lucida. It leaves behind a very smooth surface, which is ideal for PRK. It also allows for an intact epithelial flap to be lifted as a sheet from the corneal surface and hence is ideally suited for the LASEK technique. PMID- 12556377 TI - The effects of proinflammatory cytokines on cytokine-chemokine gene expression profiles in the human corneal endothelium. AB - PURPOSE: To determine the effects of proinflammatory cytokines on differential gene expression profiles in the human corneal endothelium (HCE), by using a cDNA expression array. METHODS: A human cDNA expression array technology was used to study the simultaneous expression of HCE incubated with interleukin (IL)-1alpha and tumor necrosis factor-(TNF)-alpha. Gene-specific semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were performed to examine the gene and protein expression patterns revealed by the cDNA expression array, in the presence and absence of proinflammatory cytokines. Moreover, the expression of these genes was studied in ex vivo HCE of donor cornea by RT-PCR. RESULTS: IL-1alpha and TNF-alpha upregulated the expression of 46 of 268 genes for cytokines, chemokines, and their receptors in stimulated HCE. The most upregulated genes in the cDNA expression array, those of monocyte chemotactic protein (MCP)-1 (CCL2), IL-8 (CXCL8), IL-6, and growth-related beta (GRObeta, CXCL2), were studied. Semiquantitative RT-PCR and ELISA analyses revealed the proinflammatory cytokine mediated changes in the respective gene transcription and protein expression levels. The mRNAs were detected in ex vivo HCE of donor cornea stimulated with proinflammatory cytokines. CONCLUSIONS: HCE can abundantly express cytokines and chemokines through the stimulation of proinflammatory cytokines. The detected genes, those of CCL2, CXCL8, IL-6, and CXCL2, in HCE could facilitate understanding of the inflammatory responses, including the production of keratic precipitates and the correlation between CE and an inflamed cornea or aqueous humor. PMID- 12556378 TI - Expression and function of receptors for advanced glycation end products in bovine corneal endothelial cells. AB - PURPOSE: The corneal endothelium is a target of the aging process. This study was undertaken to reveal the relationship between corneal endothelial cell (CEC) death and the accumulation of advanced glycation end products (AGEs), by investigating the possible mechanism of accumulation of AGE in CECs and its effects on CEC death. METHODS: First, the in vivo expression of the receptor was investigated for AGE (RAGE) and galectin-3, both receptors for AGE, at both the mRNA and protein levels. Second, AGEs were added to the culture media of the cultured CECs, and the uptake of AGEs, the generation of reactive oxygen species, and the induction of apoptosis were investigated. RESULTS: Immunohistochemistry and RT-PCR demonstrated that both RAGE and galectin-3 were expressed in bovine CECs. After administration of AGE-modified bovine serum albumin to the culture medium, uptake of AGE was observed in the cytoplasm of the cultured bovine CECs. In addition, with increasing concentration of AGEs, the generation of reactive oxygen and the number of apoptotic cells also increased. CONCLUSIONS: These results show that the accumulation of AGEs in CECs induced apoptosis, in part, by increasing cellular oxidative stress. The accumulation of AGEs in the CECs of elderly patients may be involved in the loss of CECs during the aging process. PMID- 12556379 TI - Nociceptive sensation and sensitivity evoked from human cornea and conjunctiva stimulated by CO2. AB - PURPOSE: To compare sensation and sensitivity evoked from human cornea and conjunctiva stimulated by CO2. METHODS: Twenty healthy participants were recruited for the study. Central corneal and temporal conjunctival chemical sensation and sensitivity of only one eye of each subject were evaluated. Air mixed with different concentrations of CO(2) was delivered by a modified Belmonte pneumatic esthesiometer. The ascending method of limits was used to determine the sensitivity and subjects were required to characterize the sensation at threshold. RESULTS: The sensations evoked by CO(2) in the cornea and conjunctiva were stinging or burning. The sensation evoked by mechanical stimulation was that of irritation. The corneal and conjunctival chemical thresholds were 31% +/- 2% and 54% +/- 5% CO(2) (mean +/- SE), respectively. The corneal and conjunctival mechanical thresholds were 80 +/- 6 and 140 +/- 10 mL/min (mean +/- SE), respectively. The corneal sensitivity was significantly higher for both mechanical and chemical stimuli (P < 0.05). CONCLUSIONS: The results suggest that CO(2) stimulates similar corneal and conjunctival nociceptors in that the interpretations were the same (i.e., nociceptive). The central cornea had a higher sensitivity to CO(2) than the temporal conjunctiva, which may reflect a different peripheral innervation, such as different nerve density or different receptor characteristics. Sensations evoked by mechanical and chemical stimulation were different, which suggests that at the peripheral level, the two modalities stimulate two different kinds of molecular receptors or channels and that this information is somehow retained within the nociceptive system. PMID- 12556380 TI - Tear evaporation dynamics in normal subjects and subjects with obstructive meibomian gland dysfunction. AB - PURPOSE: To test a newly developed tear evaporimetry system that detects real time changes in tear evaporation rates and shows the tear film stability in patients with obstructive meibomian gland dysfunction (MGD). METHODS: A ventilated chamber system with high-sensitivity microbalance sensor was used to evaluate tear evaporation. Tear evaporation rates and dynamic changes in them in response to blinking ("flip heights") were measured. Both were compared in 38 eyes of 22 normal subjects and 32 eyes of 21 patients with obstructive MGD, in a prospective case-control study. The relationship between tear evaporation rates and flip heights to meibomian gland orifice obstruction was also analyzed. RESULTS: Changes in tear evaporation rates produced by blinking were detected. The tear evaporation rates in the patients' group were 5.8 +/- 2.7(10(-7)) g/cm(2) per second, significantly higher than in normal subjects (4.1 +/- 1.4[10( 7)] g/cm(2) per second; P = 0.0008). The flip heights in the obstructive MGD group were 0.58 +/- 0.33(10(-7)) g/cm(2) per second, significantly higher than in normal subjects (0.39 +/- 0.27[10(-7)] g/cm(2) per second, P = 0.02). The correlations between both tear evaporation rates and flip heights to the meibomian gland orifice obstruction score were statistically significant (P < 0.0001 and P = 0.004, respectively). CONCLUSIONS: This new system was helpful in differentiating MGD patients from normal subjects. These significantly higher evaporation rates and higher flip heights reflect the unstable tear evaporation and may well indicate unstable tear film in patients with obstructive MGD with abnormal evaporative tear loss. PMID- 12556381 TI - HGF, MAPK, and a small physiological electric field interact during corneal epithelial cell migration. AB - PURPOSE: To investigate the effects of hepatocyte growth factor (HGF) and a small applied electric field (EF) on corneal epithelial cell (CEC) migration. METHODS: Primary cultures of bovine CECs were exposed to an EF (25-250 mV/mm) in the presence or absence of HGF (100 ng/mL). The rate and directionality of CEC migration were quantified. The expression of HGF receptors (HGFRs), p42/44 mitogen-activated protein kinase (MAPK) and the time-course of activation of p42/44 MAPK were investigated by confocal microscopy and Western blot analysis. RESULTS: CECs migrated significantly faster in the presence of an EF, HGF, or HGF and an EF combined. The distribution of HGFRs was intracellular and in the presence of an EF was concentrated in the cathode-facing side. This EF-induced asymmetrical accumulation of HGFRs correlated with the direction of CEC migration. The application of HGF or an EF led to the activation of the MAPK signaling pathway and in the presence of an EF, activation of MAPK was greater in the cathode-facing half of the CECs. Inhibition of the MAPK signaling pathway by PD98059 (100 micro M) reduced the ability of HGF and an EF to enhance the rate of CEC migration, but did not alter EF-induced cathodal directionality. CONCLUSIONS: These data suggest that both HGF and an EF augment the rate of CEC migration through activation of p42/44 MAPK. Moreover, EF-induced redistribution of HGFRs and asymmetry of MAPK signaling, although not instrumental in directing CEC migration cathodally, may be important for the signaling and maintenance of migration. PMID- 12556382 TI - Neonatal corneal stromal development in the normal and lumican-deficient mouse. AB - PURPOSE: The purpose of this study was to characterize temporally stromal growth and transparency in lumican-deficient and normal neonatal mice. METHODS: Lumican deficient mice and CD1 wild-type mice were evaluated by in vivo confocal microscopy through-focusing (CMTF) to quantify stromal and epithelial thickness and corneal light-scattering and by laser scanning CM to determine density of keratocytes from 1 day to 12 weeks after birth. RESULTS: CD1 corneas showed a rapid loss of light-scattering, decreasing by 50% from day 1 to day 12, that paralleled a 60% decrease in density of keratocytes. By contrast, the stroma demonstrated a marked swelling from day 8 to day 12, followed by thinning at day 14. Compared to corneas from CD1 mice, lumican-deficient corneas showed significantly increased (P < 0.05) light-scattering beginning at week 3 that remained elevated above wild-type levels for the duration of the study. Stromal development was also markedly altered, with thinning detected at week 3, followed by no detectable stromal growth for the duration of the study. Density of keratocytes was significantly increased, but the total cell number was similar compared with that in the wild-type cornea, suggesting no effect on keratocyte differentiation. CONCLUSIONS: Development of normal neonatal corneal transparency appears related to changes in density of keratocytes. The stroma, however, undergoes a marked swelling and thinning at the time of eyelid opening (days 8 14). In the lumican-deficient mouse, stromal swelling is abolished, indicating that this critical phase in stromal development is lumican dependent and essential for normal stromal growth and maintenance of stromal transparency. PMID- 12556383 TI - Centripetal movement of corneal epithelial cells in the normal adult mouse. AB - PURPOSE: To study the natural movement of corneal epithelial cells in the normal adult mouse with histology and in vivo microscopy. METHODS: A transgenic mouse line that was engineered to ubiquitously express green fluorescent protein (GFP) was used to visualize corneal epithelial cells. For histology, epithelial GFP was imaged in a wholemounted cornea en face, and also in frozen cross-sections, under a fluorescence microscope. For in vivo studies, the anesthetized mouse was placed on a custom-made observation platform under a fluorescence microscope. Epithelial fluorescence was digitally recorded two to three times a week, and a rate of cell movement was determined from the time-lapse sequences. RESULTS: The GFP expression in the corneal epithelium was nearly ubiquitous up to about 1 week after birth, and thereafter it gradually became sporadic, resulting in a mosaic pattern of GFP positive cells, with the brightest cells present in the basal and suprabasal layer of the epithelium. Both high- and low GFP-cells formed radial streaks toward the central cornea, frequently displaying vortex patterns at the center. Clusters of several high-GFP cells were tracked in living mice for up to 7 weeks, and an analysis of time-lapse sequences revealed that they moved centripetally at an average rate of 26 micro m/d. CONCLUSIONS: Corneal epithelium of adult GFP mice exhibits a pattern of GFP expression that is suitable for studying cell movement in the normal cornea. Epithelial cells at the basal or suprabasal layers move centripetally in these mice at an average rate of 26 micro m/d. PMID- 12556384 TI - Patterns of mucin adherence to contact lenses. AB - PURPOSE: Contact lens wear alters the preocular fluid through factors that include tear deposits on the lens. In the current study, lens-adherent material was extracted to assess whether contact lenses sample mucins from the preocular fluid. METHODS: Discarded extended-wear contact lenses were collected from patients with no ocular surface disease. Mucins were extracted in guanidine hydrochloride (GuHCl) with protease inhibitors. After the supernatant was removed, the extraction was repeated twice with the addition of 10 mM dithiothreitol, making a total of three extractions. Mucins were isolated by cesium chloride (CsCl) gradient centrifugation and size fractionated on Sepharose CL2B. Charge distribution was analyzed on ion-exchange chromatography with a lithium perchlorate (LiClO(4)) gradient. RESULTS: Contact lens-adherent mucins comprised soluble mucins and mucins that required solubilization by (repeated) dithiothreitol treatment. MUC1, MUC4, MUC2, and MUC5AC mucins eluted mainly at low buoyant densities in extractions from lenses worn long term without disinfection and at successively higher buoyant densities from monthly disposable contact lenses. Mucins with little negative charge, which were observed in all extractions, and very highly negatively charged species, present in the second and third extractions from contact lenses, had no equivalents in tissue-extracted mucins. CONCLUSIONS: Mucins adhering to contact lenses are altered forms of intracellular mucins. Different degrees of adherence of mucins to contact lenses may occur, either because of mucin characteristics or after mucin complexation with adherent materials. In the context of good contact lens hygiene, their presence may offer some protection from toxicants in the tear film, because mucins could function as acceptors for charged moieties such as free radicals. PMID- 12556385 TI - Overexpression of Insulin-like growth factor-binding protein-2 in pterygium body fibroblasts. AB - PURPOSE: To examine the expression pattern of insulin-like growth factor-binding protein (IGFBP)-2 in cultured primary pterygium fibroblasts and compare it with expression in normal conjunctival fibroblasts. METHODS: Profile of gene expression by normal conjunctival and primary pterygium fibroblasts was performed by using a cDNA microarray. The overexpression of IGFBP-2 thus identified was further confirmed by RT-PCR and Western blot analysis of cultured cells and by immunohistochemistry on primary pterygium and normal conjunctival tissue sections. RESULTS: A dramatically increased expression of IGFBP-2 mRNA was demonstrated in cDNA microarray membranes from two different pterygium fibroblasts. This finding was confirmed by RT-PCR in four additional different pterygium fibroblasts and by Western blot analysis of their culture supernatants. Immunohistochemistry of frozen sections from primary pterygium demonstrated increased staining in extracellular matrix of the stroma, compared with that of the normal conjunctiva. IGFBP-2 was also found in goblet cells of both normal conjunctival and pterygium epithelia. CONCLUSIONS: The increased expression of IGFBP-2 mRNA and protein in pterygium fibroblasts is further strong evidence to support the transformed phenotype of these cells and helps explain why there is increased growth of fibrovascular tissue. This phenotype may be used as a marker to assess the malignant nature of pterygium growth and recurrence. PMID- 12556386 TI - The corneal stroma is endowed with a significant number of resident dendritic cells. AB - PURPOSE: Dendritic cells (DCs) comprise a system of highly efficient antigen presenting cells (APCs) that initiate immune responses. The purpose of this study was to examine the normal stroma for the presence of DCs and other bone marrow (BM)-derived cells. METHODS: Normal uninflamed corneas of BALB/c and other murine strains were excised, and immunofluorescence single- and double-staining for multiple markers was performed for examination by confocal microscopy. Corneal buttons were placed in culture and immunocytochemistry and flow cytometry performed. RESULTS: MHC class II(+)CD80(+)CD86(+) cells were found in the periphery of the anterior normal stroma. These cells were CD45(+), CD11c(+)CD11b(+) suggesting a BM-derived and monocytic DC lineage. In a surprising finding, significant numbers of MHC class II(-)CD80(-)CD86(-) cells were found in the center of the anterior stroma. These cells were also CD45(+)CD11c(+)CD11b(+) but CD3(-), GR-1(-), keratan sulfate(-), and CD8alpha(-), reflecting an immature precursor phenotype of myeloid DC. In addition to DC subsets in the anterior stroma, a CD11c(-)CD11b(+) population of BM-derived cells was found primarily in the posterior stroma, representing monocytes/macrophages. These cells were rarely present in the anterior third of the normal stroma. Further, CD14(+) precursor-type DCs were found throughout the stroma. These in vivo findings were not strain specific and were confirmed by immunocytochemistry and flow cytometry analyses of cells derived from corneal explants and by transmission electron microscopy. CONCLUSIONS: This study demonstrates that, in addition to the known Langerhans cells in the corneal epithelium, at least three BM-derived cell subsets reside in the normal corneal stroma. PMID- 12556387 TI - Sustained inhibition of corneal neovascularization by genetic ablation of CCR5. AB - PURPOSE: To determine whether genetic ablation of the CC chemokine receptor CCR5 (involved in leukocyte and endothelial chemotaxis) inhibits the development of corneal neovascularization. METHODS: Wild-type C57BL/6J mice and species-specific counterparts with targeted homozygous disruption of the CCR5 gene underwent chemical and mechanical denudation of corneal and limbal epithelium. Corneas were harvested 2 and 4 weeks after injury. Neovascularization was quantified by CD31 immunostaining. Expression of VEGF protein was quantified by ELISA. RESULTS: The mean percentages of neovascularized corneal area in control mice and CCR5 deficient mice 2 weeks after denudation were 58.3% and 38.5% (P = 0.05), respectively. At 4 weeks after denudation, the corresponding percentages were 67.6% and 44.0% (P = 0.028). In CCR5-deficient mice, VEGF protein levels were reduced 51.1% at 2 weeks (P = 0.05) after injury and 37.3% at 4 weeks (P = 0.03). CONCLUSIONS: CCR5-deficient mice showed a persistent 34% to 35% inhibition of corneal neovascularization for up to 4 weeks. This inhibition correlates with reduced expression of VEGF. These data implicate CCR5 as one essential component in the development of corneal neovascularization. PMID- 12556388 TI - Serial analysis of gene expression in the corneal endothelium of Fuchs' dystrophy. AB - PURPOSE: To compare the gene expression profiles of normal human corneal endothelium with Fuchs' corneal endothelium, by using serial analysis of gene expression (SAGE). METHODS: Three pairs of normal human corneas were obtained from eye banks. Thirteen bisected Fuchs' corneal buttons were processed at the time of corneal transplantation. The endothelia of normal and Fuchs'-affected corneas were stripped, and total RNA was isolated. Serial analysis of gene expression (SAGE) was performed to identify and quantify gene transcripts. Genes over- and underexpressed by Fuchs' endothelium were limited to P < 0.01 by the method of Audic and Claverie. RESULTS: A total of 19,136 tags were identified with 9,530 from normal and 9,606 from Fuchs' endothelium. The expression of 18 transcripts was upregulated, and 36 transcripts were downregulated in Fuchs' endothelium compared with normal tissue. Upregulated transcripts included serum amyloid A1 and A2, metallothionein, and apolipoprotein D. Of the downregulated transcripts, 26 matched known genes, 3 matched expressed sequence tags (ESTs), and 7 were unknown to current databases. One downregulated transcript involved a newly reported bicarbonate transporter. Decreased transcripts related to antioxidants and proteins conferring protection against toxic stress were noted in Fuchs' versus normal endothelium including nuclear ferritin, glutathione S transferase-pi, and heat shock 70-kDa protein. Nine different SAGE tags matching mitochondrial sequences accounted for 25% of the ESTs that were decreased in Fuchs' endothelium. CONCLUSIONS: SAGE analysis comparing normal to Fuchs' endothelium demonstrates diminished expression of mitochondrial, pump function, and antiapoptotic cell defense genes. PMID- 12556389 TI - Effects of TMS over the right prefrontal cortex on latency of saccades and convergence. AB - PURPOSE: The prefrontal cortex (PFC) is known to inhibit unwanted saccades through its connections to the superior colliculus (SC). Indeed, transcranial magnetic stimulation (TMS) of the PFC decreases saccade latency by increasing the rate of express saccades. This study examined whether a similar phenomenon exists for vergence. METHODS: In a gap paradigm, six healthy subjects were asked to look at LEDs placed in a horizontal plane and to make lateral saccades, pure convergence along the median plane, and combined eye movements. Eye movements were recorded binocularly. TMS was applied over the right (r)PFC synchronously with the onset of the target. In a control condition, TMS was applied over the motor cortex (MC). RESULTS: TMS over the MC had no effect on the latency of any type of eye movements. In contrast, TMS over the rPFC (1) decreased significantly (P = 0.00367) the latency of contralateral pure saccades, (2) had no effect on the latency of pure convergence, (3) and caused a mild decrease in the latency of both the saccadic and the convergence components of combined eye movements, and the effect was bilateral. Decreased latencies were mainly due to an increase of the rate of express movements. CONCLUSIONS: The inhibition exerted by PFC over SC and preventing express movements from occurring is presumably a saccade-specific mechanism. When the saccade is combined with convergence, the express triggering can be transferred to a certain extent to the convergence. PMID- 12556390 TI - Electrophysiological and psychophysical differences between early- and late-onset strabismic amblyopia. AB - PURPOSE: To compare visual evoked potentials (VEPs) and contrast sensitivity in adults with early- or late-onset strabismic amblyopia. METHODS: Twelve adults with early- and 12 with late-onset strabismic amblyopia with similar ranges of visual acuity were studied. Pattern-onset VEPs to 30-minute checks were recorded at a range of contrast levels. Contrast sensitivity (CS) was measured at 3.2 cyc/deg using a two-alternative, forced-choice staircase method. RESULTS: There was no significant difference in VEP CII latency or amplitude between amblyopic and fellow eyes across all contrast levels for the early-onset group, but in the late-onset group, CII latencies were significantly longer and amplitudes smaller in the amblyopic eye. CII responses in both amblyopic and fellow eyes of the early-onset amblyopes were of significantly shorter latency and smaller amplitude than normal. In the late-onset group the CII responses from the amblyopic eye were of significantly increased latency and reduced amplitude compared with normal, whereas latency and amplitude of fellow eye responses did not differ significantly from normal. Late-onset amblyopes showed reduced CS across the central field for the amblyopic eye, but increased CS for the fellow eye compared with normal. In the early-onset group, central CS did not differ between amblyopic and fellow eyes or from normal. CONCLUSIONS: There are significant differences in the electrophysiological and psychophysical characteristics of adults with early- and late-onset strabismic amblyopia. PMID- 12556391 TI - Ultrasound of the horizontal rectus muscle insertion sites: implications in preoperative assessment of strabismus. AB - PURPOSE: To determine the reproducibility and accuracy of ultrasonographic (US) measurements of distances between the corneoscleral limbus and the insertion site of the medial (MR) or lateral rectus (LR) muscle compared with intraoperative measurements in patients with strabismus. METHODS: One eye in each of 17 patients with postoperative secondary strabismus caused by over- or undercorrected eso- and exotropia and one eye of each of 19 patients with previously untreated strabismus underwent five repeated measurements by high-resolution ultrasonography of the distance from the corneoscleral limbus to the muscle insertion and three actual intraoperative measurements. Reproducibility, expressed as the coefficient of variation, accuracy (relative error) of US measurements, and the agreement with intraoperative measurements were assessed. RESULTS: The coefficients of variation for US data were 2.02% and 3.18% for the MR and 7.33% and 11.77% for the LR, in the surgical and untreated groups, respectively. The relative error was 6.15% +/- 8.14% (mean +/- SD) and 3.66% +/- 12.83% for the MR muscle, and 12.21% +/- 10.66% and -7.69% +/- 7.83% for the LR muscle, in the surgical and untreated groups, respectively. The 95% limits of agreement (mean +/- 2 SD) between the US and intraoperative measurements were 0.65 +/- 1.82 and 0.15 +/- 1.42 mm for the MR muscle, and 1.12 +/- 2.01 and -0.49 +/- 0.98 mm for the LR muscle, in the surgical and untreated groups, respectively. CONCLUSIONS: The results indicate good reliability and accuracy of US readings and suggest a potential usefulness in preoperative assessment of patients with strabismus with surgical failure and missing preoperative clinical data. PMID- 12556392 TI - Deformation of the lamina cribrosa and anterior scleral canal wall in early experimental glaucoma. AB - PURPOSE: To test the hypothesis that pathophysiologic deformation of the lamina cribrosa and anterior scleral canal wall underlies the onset of confocal scanning laser tomography (CSLT)-detected optic nerve head (ONH) surface change in early experimental glaucoma. METHODS: Both eyes of four normal (two normal eyes) monkeys and four with early glaucoma (one eye with laser-induced IOP elevation, observed until the onset of CSLT-detected ONH surface change) were enucleated immediately after death and immersion fixed at IOP 0 mm Hg. In an additional four normal monkeys and five with early glaucoma, both eyes were cannulated, and IOP set to 10 mm Hg in one normal eye and either 30 or 45 mm Hg in the other (normal or early-glaucoma) eye. After 15 to 80 minutes of acute IOP elevation, these nine monkeys were perfusion-fixed. Within images of serial sagittal sections of the ONH tissues in all 17 monkeys, anterior lamina cribrosa position, laminar thickness, and scleral canal diameter were measured. For each parameter, differences between the two eyes of each monkey and between treatment groups were assessed by ANOVA. RESULTS: Within the eyes of the eight monkeys with IOP 0 mm Hg, the lamina cribrosa was posteriorly displaced and thicker and the scleral canal was enlarged at Bruch's membrane and at the anterior laminar insertion in the early-glaucoma eyes compared with the contralateral normal eyes (plastic deformation). Within the high-IOP normal eyes, the lamina cribrosa was posteriorly displaced compared with that in the low-IOP normal eyes, but there were no significant differences in laminar thickness or scleral canal diameter (normal compliance). Within the high-IOP early-glaucoma eyes, the lamina cribrosa was posteriorly displaced and thicker and the scleral canal enlarged, compared with both low-IOP normal eyes and high-IOP normal eyes (hypercompliant deformation). Differences in laminar position between the high-IOP early-glaucoma eyes and the contralateral low-IOP normal eyes (hypercompliant plus plastic deformation) were more than eight times greater than the differences between the high-IOP normal eyes and the contralateral low-IOP normal eyes (normal compliance). CONCLUSIONS: Both plastic (permanent) and hypercompliant deformation of the lamina cribrosa and anterior scleral canal wall are present in young adult monkey eyes with early experimental glaucoma. These findings suggest that damage to the ONH connective tissues occurs early in the monkey model of experimental glaucoma. PMID- 12556393 TI - Early glial responses after acute elevated intraocular pressure in rats. AB - PURPOSE: To study the responses of glial cells to a short-term elevation in intraocular pressure (IOP) in rats. METHODS: Adult Sprague-Dawley albino rats, 45 to 55 days old, were given India ink intracamerally. After 7 days, 200 spots of laser burn over 360 degrees were delivered by an Argon laser (620-637 nm; 200 mW; 200 mm; 0.2 seconds) aimed at the ink deposits in the trabecular meshwork. IOP was recorded and eye tissues at 12 hours and 1, 3, 5, 7, or 14 days after laser were examined by immunohistochemistry with antibodies against glial fibrillary acidic protein (GFAP), vimentin, S-100, ED1, and OX42. To evaluate neuronal loss, the number of cells in the retinal ganglion cell layer (RGCL) was counted on flat preparations of retinas at various times after elevation of IOP. RESULTS: Significant elevation of IOP from 1 to 7 days and loss of cells in the RGCL from 3 days onward were noted after trabecular laser photocoagulation. In the inner retina, there was a gradual and sustained increase in GFAP and S-100 immunoreactivity, but only a transient increase in vimentin immunoreactivity. No remarkable changes in GFAP, vimentin, and S-100 immunoreactivity were noted at the optic nerve head (ONH). ED1- and OX42-labeled cells were noted in the choroidal plexus, the parapapillary region of the optic nerve, and the ONH from 3 days onward, whereas expression in the retina was unremarkable. CONCLUSIONS: There is differential expression of glial cell markers in the retina and the ONH, with early loss of cells in the RGCL in response to the elevation of IOP. Macroglia such as astrocytes and Muller cells may be involved in the pathophysiology of retinal ganglion cell death or retinal repair, and activated microglial/phagocytic cells may play an important role in modulating the changes in the ONH that occur with the elevation of IOP. PMID- 12556394 TI - Effects of blur and repeated testing on sensitivity estimates with frequency doubling perimetry. AB - PURPOSE: To investigate the effect of blur and repeated testing on sensitivity with frequency doubling technology (FDT) perimetry. METHODS: One eye of 12 patients with glaucoma (mean deviation [MD] mean, -2.5 dB, range +0.5 to -4.3 dB) and 11 normal control subjects underwent six consecutive tests with the FDT N30 threshold program in each of two sessions. In session 1, blur was induced by trial lenses (-6.00, -3.00, 0.00, +3.00, and +6.00 D, in random order). In session 2, only the effects of repeated testing were evaluated. The MD and pattern standard deviation (PSD) indices were evaluated as functions of blur and of test order. By correcting the data of session 1 for the reduction of sensitivity with repeated testing (session 2), the effect of blur on FDT sensitivities was established, and its clinical consequences evaluated on total- and pattern-deviation probability maps. RESULTS: FDT sensitivities decreased with blur (by <0.5 dB/D) and with repeated testing (by approximately 2 dB between the first and sixth tests). Blur and repeated testing independently led to larger numbers of locations with significant total and pattern deviation. Sensitivity reductions were similar in normal control subjects and patients with glaucoma, at central and peripheral test locations and at locations with high and low sensitivities. However, patients with glaucoma showed larger deterioration in the total-deviation-probability maps. CONCLUSIONS: To optimize the performance of the device, refractive errors should be corrected and immediate retesting avoided. Further research is needed to establish the cause of sensitivity loss with repeated FDT testing. PMID- 12556395 TI - Strong associations between specific HLA-DQ and HLA-DR alleles and the tubulointerstitial nephritis and uveitis syndrome. AB - PURPOSE: To identify genetic markers for the tubulointerstitial nephritis and uveitis (TINU) syndrome by using human leukocyte antigen (HLA) genotyping. METHODS: Eighteen patients who had TINU syndrome were evaluated at three institutions. Typing of class I and II genes was performed by using DNA-based techniques. RESULTS: Significant associations were found with HLA-B14 (6/18 patients, 33.3%; control subjects, 5.5%; P = 0.0003; relative risk [RR] = 8.5), HLA-DQA1*01 (17/18 patients, 94.4%; control subjects, 46.6%, P = 0.0001; RR = 19.5), HLA-DQA1*0101 (14/18 patients, 77.8%; control subjects 22.2%; P < 0.0001; RR = 12.2), HLA-DQB1*05 (14/18 patients, 77.8%; control subjects 17.7%; P < 0.0001; RR = 16.3), HLA-DQB1*0501 (13/18 patients, 72.2%; control subjects 12.9%; P < 0.0001; RR = 17.6), HLA-DRB1*01 (14/18 patients, 77.8%; control subjects, 12.1%; P < 0.0001; RR = 25.5), and HLA-DRB1*0102 (13/18 patients, 72.2%; control subjects, 1.6%; P < 0.0001, RR = 167.1). The HLA haplotype most frequently identified in the study patients was HLA-DQA1*01/DQB1*05/DRB1*01 (13/18 patients, 72.2%). CONCLUSIONS: TINU syndrome is strongly associated with HLA-DQA1*01, HLA DQB1*05, and HLA-DRB1*01. The association with HLA-DRB1*0102 is one of the highest reported for any disease. Because these genes are in linkage disequilibrium, the role of the individual alleles is difficult to assess. Based on the results of the present study and on previously reported HLA associations in patients with TINU syndrome, the alphabeta dimer encoded by HLA DQA1*01/DQB1*05 may be particularly important in conferring risk for development of this disease. PMID- 12556396 TI - Survival in high-risk eyes of epithelium-deprived orthotopic corneal allografts reconstituted in vitro with syngeneic epithelium. AB - PURPOSE: In low-risk eyes of mice, most of the composite corneal grafts composed of syngeneic epithelium layered on allogeneic stroma and endothelium are accepted indefinitely. The study was undertaken to determine the fate of similar composite corneal grafts placed in high-risk mouse eyes. METHODS: Epithelium-deprived allogeneic corneas (C57BL/6) were reconstituted in vitro with BALB/c epithelium, and then transplanted orthotopically into high-risk eyes of BALB/c mice. Graft survival was assessed clinically and evaluated histologically. Acquisition of donor-specific delayed hypersensitivity (DH) was also assessed in recipient mice. Recipients bearing healthy composite grafts were immunized subcutaneously with injected C57BL/6 spleen cells at 2 or 8 weeks after grafting, after which the fate of the grafts was evaluated. RESULTS: Virtually all epithelium-deprived corneal allografts reconstituted in vitro with normal BALB/c corneal epithelium survived indefinitely when placed in high-risk eyes of BALB/c mice. Recipients of these composite grafts failed to acquire donor-specific DH when tested at both 2 and 8 weeks after grafting. Moreover, these recipients did not acquire the capacity to actively suppress donor-specific DH. Within 1 to 3 weeks of sensitization of recipient mice with spleen cells of donor origin, healthy composite grafts in residence for 2 or 8 weeks were rejected. CONCLUSIONS: Replacement of donor epithelium with syngeneic epithelium protects orthotopic allogeneic corneal grafts (stroma plus endothelium) placed in high-risk eyes from sensitizing their recipients and from immune-mediated rejection. Recipients of composite corneal grafts containing syngeneic epithelial layers act as though they are immunologically ignorant of the graft's presence. PMID- 12556397 TI - Impaired killing of HCMV-infected retinal pigment epithelial cells by anti-pp65 CD8(+) cytotoxic T cells. AB - PURPOSE: Host defense against infection by human cytomegalovirus (HCMV) is ensured in great part by cytotoxic CD8(+) T lymphocytes (CTLs) directed against the tegument protein pp65. The hyperimmediate release of incoming pp65 into the major histocompatibility complex (MHC) class I pathway after fusion of the virus with the cell membrane provides a very early mechanism of defense. In retinal pigment epithelial (RPE) cells HCMV is known to enter through endocytosis. This study was conducted to determine whether this means of penetration into the cells would allow the virus to elude immune surveillance. METHODS: Infection of RPE cells with HCMV AD169 was performed for 6 hours, 48 hours, and 8 days. Expression of intracellular pp65 in RPE cells and in the astrocytoma reference cell line U373MG was evaluated by flow cytometry, fluorescence microscopy, and Western blot analysis. Killing of both HCMV-infected cell lines by HLA-A2-restricted CD8(+) CTLs directed against pp65 was monitored by (51)Cr-release assays. RESULTS: RPE cells were not lysed by CTLs directed against incoming pp65, contrary to U373MG. Moreover, both cell lines were not killed by anti-pp65 CTLs later after infection, because of the MHC class-I-downregulating effect of HCMV unique short (US2-11) proteins. CONCLUSIONS: In RPE cells, both HCMV entry through endocytosis and the immunosuppressive effect of US proteins could allow the virus to evade immune surveillance at any stage of infection, which could promote viral spreading within the retina. PMID- 12556398 TI - Activation of metallothioneins and alpha-crystallin/sHSPs in human lens epithelial cells by specific metals and the metal content of aging clear human lenses. AB - PURPOSE: To identify those metallothionein and alpha-crystallin/small heat-shock genes induced by toxic metals in human lens cells and to evaluate the levels of these metals between young and aged human lenses. METHODS: Human SRA01/04 and primary human lens epithelial cells were cultured and exposed to Cd(2+), Cu(2+), and Zn(2+). The levels of lens metallothioneins (Ig, If, Ih, Ie, and IIa) and alpha-crystallin/small heat-shock (alphaA-crystallin, alphaB-crystallin, and HSP27) genes were analyzed by semiquantitative and quantitative competitive RT PCR. The content of aluminum, cadmium, calcium, chromium, copper, iron, lead, magnesium, manganese, nickel, potassium, sodium, and zinc in young (mean, 32.8 years), middle-aged (mean, 52.3 years), and old (mean, 70.5 years) human lenses was analyzed by inductively coupled plasma-emission spectroscopy. RESULTS: Lens metallothioneins (Ig, If, Ih, Ie, and IIa) and alpha-crystallin/small heat-shock genes (alphaA-crystallin, alphaB-crystallin, and HSP27) were differentially induced by specific metals in SRA01/04 human lens epithelial cells. Cd(2+) and Zn(2+), but not Cu(2+), induced the metallothioneins, whereas Cd(2+) and Cu(2+), but not Zn(2+), induced alphaB-crystallin and HSP27. alphaA-crystallin was induced by Cu(2+) only. Similar responses of the metallothionein IIa gene were detected in identically treated primary human lens epithelial cells. Cd(2+) and Zn(2+) induced metallothionein IIa to five times higher levels than metallothionein Ig. Of 13 different metals, only iron was altered, exhibiting an 81% decrease in old versus young lenses. CONCLUSIONS: Induction of metallothioneins and alpha-crystallin/small heat shock proteins by different metals indicates the presence of metal-specific lens regulatory pathways that are likely to be involved in protection against metal-associated stresses. PMID- 12556400 TI - Mechanical properties of the human posterior lens capsule. AB - PURPOSE: To investigate mechanical properties of the human posterior lens capsule. METHODS: Twenty-five human donor eyes were obtained from an eye bank. The age of the donors ranged from 1 to 94 years. Test specimens were prepared as tissue rings from posterior lens capsules by means of excimer laser. Capsular thickness was measured microscopically as the difference in focus between microspherules placed on the outer and inner surfaces of the capsule. The capsular rings were slipped over two pins connected to a motorized micropositioner and a force transducer and stretched at a constant rate with continuous recording of load and deformation. Data for the posterior lens capsule were compared with previously published data for the anterior lens capsule. RESULTS: The thickness of the posterior lens capsule ranged from 4 to 9 micro m and showed no significant changes with age. Ultimate mechanical strength of the posterior lens capsule decreased significantly with age. Ultimate strain ranged from 101% to 34%, ultimate load ranged from 15.9 to 1.1 mN, ultimate stress ranged from 16.1 to 1.1 N/mm(2), ultimate elastic stiffness ranged from 52.1 to 5.7 mN, and ultimate elastic modulus ranged from 27.4 to 3.3 N/mm(2). The load strain and the stress-strain relationships in the posterior lens capsule were nonlinear, and therefore elastic stiffness and elastic modulus varied as a function of strain. In the low-strain region (0%-10% strain), elastic stiffness and elastic modulus ranged between 0.3 to 2.4 mN and 0.3 to 2.3 N/mm(2), respectively, and seemed to increase during the first part of life until middle age. CONCLUSIONS: Mechanical strength of the posterior lens capsule was found to decrease markedly with age. The age-related loss of mechanical strength seemed to begin earlier in the posterior lens capsule than in the anterior lens capsule. In accommodative function range (low strains), the mechanical quality of the posterior lens capsule was similar to the anterior lens capsule, which indicates that the mechanical effectiveness of the lens capsule in situ varies proportionally with capsular thickness. PMID- 12556399 TI - Evaluation of fibroblast growth factor signaling during lens fiber cell differentiation. AB - PURPOSE: Previous studies have implicated members of the fibroblast growth factor (FGF) and insulin-like growth factor (IGF) families as stimulators of lens fiber cell differentiation in rodent and chicken embryo lenses, respectively. In the present study, the role of FGFs in fiber cell differentiation and epithelial cell proliferation in chicken embryos was examined. METHODS: Lenses were injected on embryonic day (E)3 with replication-defective retroviruses that express full length or truncated FGF receptor (FGFR)-1 or a secreted form of FGF1. Lens epithelial explants were cultured in defined medium or medium supplemented with FGFs or vitreous humor, in the presence or absence of the FGF receptor antagonist SU5402. Explants were also cultured in vitreous humor that had been depleted of heparin-binding growth factors. Cell elongation was measured optically and protein accumulation by densitometry and Western blot analysis. RESULTS: Lens fiber cell differentiation was not inhibited in cells infected with virus expressing truncated FGFR1. Epithelial cells infected with virus encoding a secreted form of FGF1 did not differentiate into ectopic fiber cells. Viral transduction of FGFR1, truncated FGFR1, or FGF1 did not appreciably alter the proliferation of lens epithelial cells. Bovine vitreous humor stimulated chicken embryo lens epithelial cells to elongate and express markers of lens fiber cell differentiation. Bovine vitreous humor, but not FGF2, protected lens epithelial cells from apoptosis. Depleting vitreous humor of heparin-binding growth factors or treatment of lens cells with SU5402 did not inhibit the initial, rapid phase of lens cell elongation. Both treatments, used separately or together, reduced but did not prevent the expression of later markers of fiber cell differentiation. CONCLUSIONS: Fiber differentiation factors that are not members of the FGF family are present in chicken and mammalian vitreous humor. The factors that stimulate fiber cell differentiation in avian and mammalian eyes are similar. PMID- 12556401 TI - Activation of scleral cold thermoreceptors by temperature and blood flow changes. AB - PURPOSE: To study the response of scleral cold receptors located in areas of the eye unexposed to temperature and blood flow changes. METHODS: In anesthetized cats, the neural activity was recorded from single, cold-sensory fibers of the ciliary nerves innervating the sclera and limbus. Controlled temperature changes of the receptive field were performed with a contact thermode. Ocular blood flow reductions were obtained by occluding the ipsilateral common carotid artery for 30 to 60 seconds with a compressor placed around the artery. Local blood flow was measured with a laser Doppler flowmeter. Temperature was measured with a microprobe introduced in the subscleral space. Ocular sympathetic stimulation was performed with a pair of silver electrodes placed on the preganglionic cervical sympathetic trunk. To induce local hypoxia, N(2) was applied on the scleral surface with a specially designed chamber. For systemic hypoxia the breathing air was replaced with a gas mixture containing 10% O(2) in N(2). RESULTS: Sensory nerve fibers identified as cold receptors exhibited ongoing nerve activity in bursts at 35 degrees C and responded to cooling pulses applied to their receptive fields with an increase in the impulse discharge that reached a peak and decayed gradually to a lower level. When temperature was reduced from 35 degrees C to 34 degrees C, frequency increased monotonically with decreasing temperature of the sclera. Between 35 degrees C and 30 degrees C, peak and mean frequencies were roughly proportional to temperature of the sclera. The characteristics of burst discharges also depended on scleral temperature. Electrical stimulation of the cervical sympathetic trunk induced a decrease in blood flow and temperature and evoked an increase in the firing frequency of cold-sensory fibers that was proportional to the frequency of stimulating pulses. Carotid occlusion also elicited an increase of the discharge of cold thermoreceptor fibers that occurred in parallel with a decrease in blood flow and temperature in the receptive field area. Local or systemic hypoxia did not modify appreciably the spontaneous firing frequency of scleral cold-sensory fibers. CONCLUSIONS: Scleral and episcleral cold-sensory fibers encoded as a change in their impulse frequency and firing pattern temperature reductions of less than 1 degrees C in scleral tissues. Activation of scleral and episcleral cold-sensory fibers by sympathetic vasoconstriction and acute arterial pressure reductions appear to be secondary to the temperature decrease that accompanies the reduction in ocular blood flow caused by these maneuvers. Scleral thermosensory fibers are located in ocular territories not directly exposed to external temperature changes. Thus, the sensory information on local blood flow variations provided by these receptors may be involved in a reflex regulation of choroidal blood flow that functions to maintain a constant temperature and blood supply to the retina. PMID- 12556402 TI - Cell membrane stretch modulates the high-conductance Ca2+-activated K+ channel in bovine trabecular meshwork cells. AB - PURPOSE: Anterior chamber structures are subjected to changes in intraocular pressure (IOP). Several studies have pointed out that trabecular meshwork (TM) cells are sensitive to mechanical stretch and that cell-signaling mechanisms are activated in response to elevated pressure. Because membrane stretch has been shown to be a modulator of several ionic conductances, this study was conducted to determine its effects on the high-conductance Ca(2+)-activated K(+) (BK(Ca)) channels present in TM cells. METHODS: Primary cultures of TM cells from bovine eyes were used. Patch-clamp recordings were performed in the cell-attached, inside-out, and whole-cell configurations. To stretch the cell membrane, both suction to the rear end of the patch pipette and hypotonic shock were used. Intracellular calcium concentration ([Ca(2+)](i)) was measured in TM cells loaded with fura-2, using an epifluorescence microscope coupled to a charge-coupled device (CCD) camera. RESULTS: Electrophysiological characterization of BK(Ca) channels was in agreement with previous studies. In cell-attached patches, the open probability of the BK(Ca) channel (i.e., the amount of time the channel is open) increased consistently when 14- to 45-mm Hg suctions were applied at a constant depolarized voltage. At a constant pressure (25 or 45 mm Hg), channel openings increased when depolarizing pulses were applied to the patch. Stretch activation of the BK(Ca) channel was not mediated by increases in [Ca(2+)](i), because it was present in inside-out patches maintained at a constant Ca(2+) concentration. Nevertheless, it cannot be ruled out that at low suction levels, a minimum Ca(2+) concentration is necessary for channel activation. Whole-cell currents carried by BK(Ca) channels increased when the isotonic solution in the bath was exchanged with a hypotonic solution and were selectively blocked by iberiotoxin. In our conditions, the hypotonic shock did not modify [Ca(2+)](i). CONCLUSIONS: The data show that in TM cells, open probability of the BK(Ca) channel is enhanced by membrane stretching as well as by membrane depolarization and [Ca(2+)](i). Changes in membrane tension induced by cell volume increase also activated whole-cell BK(Ca) currents. Homeostatic mechanisms in TM cells may involve BK(Ca) channel activation in response either to changes in cell volume or changes in IOP. PMID- 12556404 TI - Noninvasive assessment of aqueous humor turnover in the mouse eye. AB - PURPOSE: To develop a noninvasive test for monitoring changes in aqueous humor turnover in the mouse eye. METHODS: After topical instillation of fluorescein, the rate of decay of fluorescence from aqueous humor and cornea was monitored in Black Swiss, C57 Bl6, and DBA 2J mice with a microscope equipped with epifluorescence and a charge-coupled device (CCD) camera. RESULTS: The rate of decay of fluorescence was identical in right and left eyes over an approximately 70-minute measurement period. The rate of decay was similar in normal mice aged 2 and 18 months. Pilocarpine and latanoprost, known to enhance aqueous humor outflow in humans, accelerated the decay of fluorescence. Levobunolol, known to inhibit aqueous humor inflow in humans, slowed decay. Dimethylamiloride, an inhibitor of the Na(+),H(+) exchanger that is known to act on cultured cells of both the ciliary epithelium and trabecular meshwork and to lower mouse intraocular pressure (IOP), enhanced decay. DBA 2J mice, in which secondary glaucoma develops, displayed a slower decay of fluorescence at 18 months of age than age-matched unaffected animals. CONCLUSIONS: Monitoring decay of fluorescence provides a noninvasive index of aqueous humor dynamics in the mouse eye that facilitates study of ocular hypotensive drugs and mouse models of glaucoma. Coupled with knowledge of IOP, it permits semiquantitative conclusions about the relative roles of aqueous humor inflow and outflow in conditions with altered IOP. Based on this approach, dimethylamiloride appears to lower mouse IOP primarily by enhancing outflow of aqueous humor. PMID- 12556403 TI - Human trabecular meshwork cell responses induced by bimatoprost, travoprost, unoprostone, and other FP prostaglandin receptor agonist analogues. AB - PURPOSE: To determine the functional agonist potencies of the intraocular pressure (IOP)-lowering prostaglandin F (FP)-class prostaglandin (PG) analogues (e.g., travoprost, latanoprost, bimatoprost, and unoprostone isopropyl ester) in human trabecular meshwork (h-TM) cells, by using phosphoinositide (PI) turnover and intracellular Ca(2+) ([Ca(2+)](i)) mobilization, and to confirm the FP nature of these receptors by using an FP receptor antagonist, 11beta-fluoro-15-epi-15 indanyl-PGF(2alpha) (AL-8810). METHODS: FP-receptor-mediated PI turnover and [Ca(2+)](i) mobilization were measured in h-TM cells by determining the accumulation of [(3)H]-inositol phosphates ([(3)H]-IPs) by anion-exchange chromatography and real-time fluorescence imaging, respectively. RESULTS: Various PG analogues concentration-dependently stimulated production of [(3)H]-IPs in h TM cells with the following agonist potencies (median effective concentration; EC(50)): travoprost acid (EC(50) = 2.4 nM) > cloprostenol (EC(50) = 4.5 nM) > (+/ )-fluprostenol (EC(50) = 10.8 nM) > latanoprost acid (EC(50) = 34.7 nM) > bimatoprost acid (EC(50) = 112 nM) > PGF(2alpha) (EC(50) = 120 nM) >> unoprostone (UF-021; EC(50) = 3280 nM) > S-1033 (EC(50) = 4570 nM; all n = 3-9). Prodrug derivatives of these compounds exhibited the following potencies: travoprost (isopropyl ester; EC(50) = 89.1 nM) > latanoprost (isopropyl ester; EC(50) = 778 nM) > bimatoprost (amide; EC(50) = 1410-6940 nM). Travoprost acid, PGF(2alpha,) unoprostone, and S-1033 were tested in addition for [Ca(2+)](i) mobilization and found to have rapid and dose-dependent effects. The FP receptor-selective antagonist AL-8810 antagonized the (+/-)-fluprostenol-induced PI turnover in these cells (K(i) = 2.56 +/- 0.62 micro M) as well as that induced by bimatoprost and acids of latanoprost and travoprost. The agonist and antagonist potencies of the PG analogues from the PI turnover assays in h-TM cells correlated well with PI turnover data obtained from the cloned human ciliary body FP receptor (r = 0.92; P < 0.0001). CONCLUSIONS: The pharmacology of the h-TM cell FP-receptor mediated PI turnover and [Ca(2+)](i) mobilization was defined using numerous synthetic (FP-selective) PG agonist analogues and an FP receptor antagonist, AL 8810. Bimatoprost, travoprost, latanoprost, unoprostone isopropyl ester, and their respective free acids were shown to be FP agonists in the h-TM cells. PMID- 12556405 TI - Role of endothelin-1 in choroidal blood flow regulation during isometric exercise in healthy humans. AB - PURPOSE: There is evidence that the choroid has some autoregulatory capacity in response to changes in ocular perfusion pressure (OPP). The mediators of this response are hitherto unidentified. The hypothesis for the current study was that endothelin (ET)-1 and/or angiotensin (ANF)-II may be involved in choroidal vasoconstriction during an increase in OPP. METHODS: To test this hypothesis a randomized, double-masked, placebo-controlled, three way crossover study was performed in 12 healthy male volunteers. Subjects received on different study days intravenous infusions of the specific ET(A) receptor antagonist BQ-123, the angiotensin converting enzyme inhibitor enalapril or placebo. During these infusion periods subjects were asked to squat for 6 minutes. Choroidal blood flow was measured using a confocal laser Doppler flowmeter and ocular perfusion pressure (OPP) was calculated from mean arterial pressure and intraocular pressure. RESULTS: BQ-123 and enalapril had no effect on basal blood pressure, pulse rate, intraocular pressure, or choroidal blood flow. During isometric exercise, a pronounced increase in mean arterial pressure paralleled by an increase in OPP was observed. Although choroidal blood flow slightly increased during squatting, the increase was much less pronounced than the increase in OPP, indicating some regulatory potential of the choroid. Enalapril did not alter the choroidal pressure-flow relationship during isometric exercise, but BQ-123 induced a significant leftward shift of the pressure-flow curve (P < 0.001). CONCLUSIONS: The present data indicate that ET-1, but not ANG II, plays a role in choroidal blood flow regulation during isometric exercise in healthy humans. Hence, impaired choroidal autoregulation in patients with ocular vascular diseases may arise from an altered endothelin system. Further studies in such patients are warranted to verify this hypothesis. PMID- 12556406 TI - Role of NO in choroidal blood flow regulation during isometric exercise in healthy humans. AB - PURPOSE: Nitric oxide (NO) is an important regulator of basal choroidal blood flow. Animal experiments indicate that NO is also involved in choroidal blood flow regulation during changes in ocular perfusion pressure and inhibition of NO synthase (NOS) has been reported to shift choroidal pressure-flow curves to the right. The hypothesis for the study was that inhibition of NOS may influence choroidal blood flow during isometric exercise. METHODS: To test this hypothesis, a randomized, double-masked, placebo-controlled, three-way crossover study was performed in 12 healthy male volunteers. Subjects received on different study days intravenous infusions of N(G)-monomethyl-L-arginine (L-NMMA), phenylephrine, or placebo. During these infusion periods, subjects were asked to squat for 6 minutes. Choroidal blood flow was assessed with laser Doppler flowmetry, and ocular perfusion pressure (OPP) was calculated from mean arterial pressure and intraocular pressure. RESULTS: L-NMMA and phenylephrine increased resting OPP by 10% and 13%, respectively, but only L-NMMA reduced resting choroidal blood flow ( 17%, P < 0.001). The relative increase in OPP during isometric exercise was comparable with all drugs administered. Isometric exercise increased choroidal blood flow during administration of placebo and phenylephrine, but not during administration of L-NMMA (P < 0.001 vs. placebo). CONCLUSIONS: These data indicate that NO plays an important role in the regulation of choroidal blood flow during isometric exercise. PMID- 12556407 TI - Biodegradable intrascleral implant for sustained intraocular delivery of betamethasone phosphate. AB - PURPOSE: To evaluate the feasibility of using a biodegradable intrascleral implant for intraocular sustained delivery of betamethasone phosphate (BP). METHODS: The intrascleral implant (0.5 mm thick and 4 mm in diameter) was made of poly(DL-lactide) containing 25% betamethasone phosphate. The in vitro release of BP from the implant was evaluated by high-performance liquid chromatography (HPLC). The implants were placed into a scleral pocket in the rabbit's eye. The concentrations of BP in the aqueous humor, vitreous, and retina-choroid were measured by HPLC. The toxicity and biocompatibility of the implant were evaluated by slit lamp examination, electroretinography, and light microscopy. RESULTS: In vitro studies demonstrated that the implants released BP in a biphasic pattern for at least 8 weeks. The BP concentrations in the vitreous and the retina choroid remained within the concentration range capable of suppressing inflammatory responses for more than 8 weeks. The BP concentration was greater in the retina-choroid than in the vitreous. In the aqueous humor, BP was below the detection limit during the observation period. No significant toxicity to the retina was observed. Also, the implant showed good biocompatibility in the eye. CONCLUSIONS: These results suggest that the intrascleral implant would be a promising system for delivery of steroid to the posterior segment of the eye. PMID- 12556408 TI - Effect of acute hyperglycemia on oxygen and oxidative metabolism in the intact cat retina. AB - PURPOSE: The Crabtree effect is the phenomenon of inhibition of respiration by glycolysis, as a result of elevated glucose levels. It is not certain whether the Crabtree effect occurs in the retina, which has a high glycolytic capacity. In the current study, in vivo photoreceptor oxygen consumption was examined during the normo- and hyperglycemic states in the dark-adapted cat retina to determine whether the Crabtree effect occurs in the outer retina. METHODS: Spatial profiles of oxygen tension were obtained in the cat retina, in vivo, with the use of oxygen microelectrodes during control conditions and acute (5.19 +/- 0.83 hour) episodes of hyperglycemia (blood glucose, >350 mg/dL). The outer retinal portions of the profiles were fitted to a model of oxygen diffusion to quantify photoreceptor oxygen consumption. RESULTS: Photoreceptor oxygen consumption did not significantly change during hyperglycemia compared with control conditions. Choroidal PO(2) decreased during hyperglycemia by an average of 5.8 +/- 7.4 mm Hg. This led to an increase in the fraction of O(2) used by the photoreceptors that was derived from the inner retina. Choroidal PO(2) did not recover when blood glucose levels were returned to normal. Average inner retinal PO(2) was not affected by the episodes of hyperglycemia. CONCLUSIONS: The Crabtree effect does not occur to any significant degree in the outer retina, because hyperglycemia did not affect photoreceptor oxygen consumption. Choroidal PO(2) decreased during hyperglycemia, and the oxygen deficit was made up by the retinal circulation. PMID- 12556409 TI - Subconjunctival doxifluridine administration suppresses rat choroidal neovascularization through activated thymidine phosphorylase. AB - PURPOSE: Doxifluridine (5'-deoxy 5-fluorouridine) is an oral anticancer drug with antiangiogenic effects, with vasoclastic action that is enhanced by a major member of the pyrimidine phosphorylases, thymidine phosphorylase (TP). Previous studies have demonstrated that TP is upregulated in the lesions where pathologic angiogenesis occurs and TP itself promotes angiogenesis. To investigate the possible role of TP and doxifluridine in choroidal neovascularization (CNV), the expression level of TP was measured and the effect of doxifluridine was investigated in rat eyes with experimental CNV. METHODS: CNV was induced in rat eyes by diode laser photocoagulation. The expression level of TP in the laser treated and control eyes was examined with high-performance liquid chromatography (HPLC). For the evaluation of CNV activity, the intensity of fluorescein leakage from the photocoagulated lesions was scored, and the areas of CNV lesions were measured histologically in the control eyes and eyes treated with a subconjunctival injection of doxifluridine 14 days after photocoagulation. RESULTS: The expression level of TP was higher in the laser-treated eyes than in the control eyes. Fluorescein leakage from the CNV lesions significantly decreased in the eyes given a subconjunctival injection of doxifluridine compared with the control. Histologic analysis demonstrated that both the areas of CNV lesions and the degree of vascular formation in the subretinal membrane were reduced in the doxifluridine-treated eyes compared with the control eyes. CONCLUSIONS: TP may be involved in the formation of CNV. Subconjunctival injection of doxifluridine significantly reduced experimental CNV activity without apparent adverse effects. These results suggest the possibility that doxifluridine can be beneficial in treating CNV. PMID- 12556410 TI - An in vivo doxycycline-controlled expression system for functional studies of the retina. AB - PURPOSE: Transgenic mice were developed that express tetracycline-controlled transactivator 1 (tTA1) specifically in photoreceptor cells. In these mice the transcription of the gene of interest can be easily inactivated in the retina in a short time frame. METHODS: A construct was prepared containing tTA1 under control of the murine rhodopsin regulatory region. This construct was used for the generation of transgenic mice. In situ hybridization was performed to study the distribution of the transactivator in the retina. The activity of the transactivator was analyzed by mating the lines with a luciferase reporter transgenic mouse. tTA1 activity and doxycycline's ability to block it were analyzed by luciferase assay. The effects of tTA1 on the retina were assessed by histology and electrophysiology. RESULTS: Two transgenic lines were developed that specifically express tTA1 in photoreceptor cells. The time course of transgene expression replicated transcription of endogenous rhodopsin. tTA1 was not toxic to the retina. Transactivator activity was blocked readily by doxycycline. CONCLUSIONS: An expression system for photoreceptor cells was generated to drive transcription in a cell-specific and time-controllable manner. This system is suitable for the study of factors involved in retinal biology and of mutant forms of genes involved in retinal diseases. PMID- 12556411 TI - Automated detection of fundus photographic red lesions in diabetic retinopathy. AB - PURPOSE: To compare a fundus image-analysis algorithm for automated detection of hemorrhages and microaneurysms with visual detection of retinopathy in patients with diabetes. METHODS: Four hundred fundus photographs (35-mm color transparencies) were obtained in 200 eyes of 100 patients with diabetes who were randomly selected from the Welsh Community Diabetic Retinopathy Study. A gold standard reference was defined by classifying each patient as having or not having diabetic retinopathy based on overall visual grading of the digitized transparencies. A single-lesion visual grading was made independently, comprising meticulous outlining of all single lesions in all photographs and used to develop the automated red lesion detection system. A comparison of visual and automated single-lesion detection in replicating the overall visual grading was then performed. RESULTS: Automated red lesion detection demonstrated a specificity of 71.4% and a resulting sensitivity of 96.7% in detecting diabetic retinopathy when applied at a tentative threshold setting for use in diabetic retinopathy screening. The accuracy of 79% could be raised to 85% by adjustment of a single user-supplied parameter determining the balance between the screening priorities, for which a considerable range of options was demonstrated by the receiver operating characteristic (area under the curve 90.3%). The agreement of automated lesion detection with overall visual grading (0.659) was comparable to the mean agreement of six ophthalmologists (0.648). CONCLUSIONS: Detection of diabetic retinopathy by automated detection of single fundus lesions can be achieved with a performance comparable to that of experienced ophthalmologists. The results warrant further investigation of automated fundus image analysis as a tool for diabetic retinopathy screening. PMID- 12556412 TI - Automated detection of diabetic retinopathy in a fundus photographic screening population. AB - PURPOSE: To evaluate the performance of an automated fundus photographic image analysis algorithm in high-sensitivity and/or high-specificity segregation of patients with diabetes with untreated diabetic retinopathy from those without retinopathy. METHODS: This was a retrospective cross-sectional study of 260 consecutive nonphotocoagulated eyes in 137 diabetic patients attending routine photographic retinopathy screening. Mydriatic 60 degrees fundus photography on 35 mm color transparency film was used, with a single fovea-centered field. Routine grading was based on visual examination of slide-mounted transparencies. Reference grading was performed with specific emphasis on achieving high sensitivity. Computer-assisted automated red lesion detection was performed on digitized transparencies. RESULTS: When applied in a screening population comprising patients with diabetes with untreated diabetic retinopathy in any eye and patients with diabetes without retinopathy, the automated lesion detection correctly identified 90.1% of patients with retinopathy and 81.3% of patients without retinopathy. A per-eye analysis for methodological purposes demonstrated that the automated lesion detection could be adapted to simulate various visual evaluation strategies. When adapted at high sensitivity, the automated system demonstrated sensitivity at 93.1% and specificity at 71.6%. When adapted at high specificity the automated system demonstrated sensitivity at 76.4% and specificity at 96.6%, closely matching routine visual grading at sensitivity 76.4% and specificity 98.3%. CONCLUSIONS: Automated detection of untreated diabetic retinopathy in fundus photographs from a screening population of patients with diabetes can be made with adjustable priority settings, emphasizing high-sensitivity identification of diabetic retinopathy or high-specificity identification of absence of retinopathy, covering opposing extremes of visual evaluation strategies demonstrated by human observers. PMID- 12556413 TI - Adeno-associated virus encoding green fluorescent protein as a label for retinal pigment epithelium. AB - PURPOSE: To determine whether transduction with adeno-associated virus encoding green fluorescent protein (AAV-GFP) is useful for labeling transplanted retinal pigment epithelial cells (RPE). METHODS: Transduction was performed by infection of confluent or subconfluent cultured feline RPE or by subretinal injection. Cells transduced in vitro were analyzed to determine label stability over time and label conservation with cell division. RPE transduced in vivo were harvested at 5 weeks for transplantation or immunohistochemical detection. Two cats received subretinal injections of harvested cells and were killed at 3 or 7 days. RESULTS: In vitro transduction of confluent RPE resulted in stable GFP fluorescence for at least 3 months. There was a marked decline in fluorescence after cell division. Nonconfluent transduced cells conserved label after cell division but showed a marked decline in the number of cells, due to cell death. In vivo transduction resulted in a high level of labeling, allowing labeled cells to be harvested and transplanted. Transplanted cells were detected immunohistochemically. Photoreceptor labeling was detected over areas containing a high density of transplanted, labeled RPE derived from cells transduced in vivo. Possible light toxicity to transduced RPE was observed. CONCLUSIONS: AAV GFP-labeling of confluent cultured RPE and RPE in situ can be used to identify transplanted RPE, with some reservations. Cell division may cause dilution of the label, and release of cell contents into the subretinal space may cause label transfer to photoreceptors. Exposure to light of transduced cells should be limited. PMID- 12556414 TI - AAV-mediated expression of vascular endothelial growth factor induces choroidal neovascularization in rat. AB - PURPOSE: To develop a small-animal model of choroidal neovascularization (CNV) by injecting adeno-associated virus (AAV)-VEGF into the subretinal space (SRS) of rats. METHODS: An adeno-associated viral vector encoding human VEGF(165) was injected into the subretinal space (SRS) of Sprague-Dawley or Long Evans rats. Expression of VEGF was identified by RT-PCR and immunohistochemistry. Physiological and pathologic changes in the retina and choroid were evaluated by electroretinography, fluorescein angiography, light microscopy, and three dimensional reconstruction of serial sections. RESULTS: Green fluorescent protein (GFP) and VEGF were expressed for at least 20 months in the retina and retinal pigment epithelium (RPE). Histologic sections showed extensive subretinal neovascularization, degenerating photoreceptors, and proliferating RPE at 5 weeks to 20 months after injection of AAV-VEGF. At 2 to 12 months after injection, leaking blood vessels were detected by fluorescein angiography. Electroretinogram a- and b-wave amplitudes were significantly decreased during this time. Three dimensional reconstruction of serial sections demonstrated that choroidal blood vessels penetrated Bruch's membrane, one of them splitting into three branches in the SRS. In the current model, CNV was produced in 95% of the animals tested (19/20). It persisted for more than 20 months, a necessary requirement for modeling the development of CNV in age-related macular degeneration (AMD). CONCLUSIONS: In this study, a highly reproducible animal model of long-lasting CNV was developed. This model is being used to test antiangiogenic molecules to reduce or inhibit CNV and could be extended to primates. PMID- 12556415 TI - Mechanism by which brain-derived neurotrophic factor increases dopamine release from the rabbit retina. AB - PURPOSE: To determine whether BDNF modulates the release of dopamine from amacrine cells in the rabbit retina. METHODS: Isolated retinas from rabbits killed with pentobarbital were incubated in Krebs bicarbonate medium containing pargyline, nomifensine, and bovine serum albumin. The medium was changed at 10 minute intervals, and the dopamine in the resultant samples measured by HPLC. Five samples were collected to establish the spontaneous resting release of dopamine, and then the retina was exposed to BDNF for a further two collection periods. Double-label immunohistochemistry was used to identify tyrosine hydroxylase containing neurons and to localize TrkB (BDNF) receptors. RESULTS: Exposure of the retina to BDNF (70-150 ng/mL) caused a concentration-dependent increase in the release of dopamine. The maximum effect was produced by 150 ng/mL BDNF, which almost doubled the release. The BDNF-evoked release was abolished in low-calcium/high-magnesium medium. It was also prevented by the tyrosine kinase inhibitors k252a and genistein, the phospholipase inhibitor U73122, and the sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitors thapsigargin and cyclopiazonic acid. Antagonists of gamma-aminobutyric acid (GABA) and glutamate did not affect the BDNF-evoked release of dopamine. ELISA assay confirmed the presence of BDNF in the retina, and immunohistochemistry revealed that some tyrosine hydroxylase-containing neurons possessed TrkB receptors. CONCLUSIONS: BDNF quickly (within minutes) increases the release of dopamine from amacrine cells in the rabbit retina by an action that is likely to involve TrkB receptors. The mechanism linking TrkB receptor activation to the release of dopamine involves activation of phospholipase-Cgamma, with the subsequent production of IP(3) and calcium release from the endoplasmic reticulum. The resultant capacitative entry of calcium seems to be the actual trigger for BDNF-induced release of dopamine. PMID- 12556416 TI - Non-neural regions of the adult human eye: a potential source of neurons? AB - PURPOSE: Because it is known that both melanocytes and neurons are generated from neural crest stem cells and their derived precursors, the current study was undertaken to evaluate whether adult human ocular tissues, containing melanocytes, have the capacity to generate neuronlike cells in vitro. METHODS: Choroid and Sclera cells from adult human eyes were separately dissociated and cultivated in the presence of epidermal growth factor and 10% fetal bovine serum. No retinal pigmented epithelial cells were detected. After cell growth, cells were transferred under conditions known to induce neuronal differentiation. Cells were plated on laminin in the presence of fibroblast growth factor-2 or brain derived neurotrophic factor. RESULTS: Cells derived from the sclera and the choroid of 15 donors proliferated to attain a 10(8)-fold increase in the number of cells within 4 months. At each passage, groups of cells differentiated into cells with neuronal morphology, expressing neuronal markers confirmed by immunocytochemistry and RT-PCR analyses, such as beta-tubulin-III, neurofilament, and tau. Parallel to neuronlike formation, glialike cells, revealed by expression of vimentin and P0, were generated in large amounts. Although, absent from choroid and sclera tissues, nondifferentiated cells appeared in cultures. CONCLUSIONS: The adult human eye conserves cells able to recapitulate certain neural developmental features. This observation opens new perspectives to study human neurogenesis and to provide an important source of neurons for transplantation studies in the retina and other regions of the central nervous system. PMID- 12556417 TI - Apical and basal regulation of the permeability of the retinal pigment epithelium. AB - PURPOSE: The functional characteristics of tight junctions in the outer blood retinal barrier change during embryonic development and in the presence of disease. A culture model of developing retinal pigment epithelium (RPE) was used to examine the regulation of the tight junctions. METHODS: RPE from chick embryos was cultured on filters that separated the apical and basal medium compartments. Cultures were maintained in various combinations of serum-free medium, serum-free medium that was conditioned by neural retinas, or serum-free medium that was supplemented with bovine pituitary extract, serum, or various hormones. Function was monitored by the transepithelial electrical resistance (TER) or the permeation of small organic tracers. Structure was monitored by immunofluorescence and freeze-fracture electron microscopy. RESULTS: Functional analysis indicated differences in permeability among RPE of different embryonic age and culture conditions. In serum-free medium, the tight junctions were leaky or failed to form. Barrier properties increased if pituitary extract was added to the basal medium chamber or retina-conditioned medium was added to the apical chamber. Retina-conditioned medium was more effective at organizing tight junctional strands into a continuous network, but bovine pituitary extract appeared to modulate the permeability of that network. In combination, they synergistically elevated the TER to physiological levels. Although the thyroid hormone T3 had no effect, serum in the apical medium chamber inhibited the ability of RPE cells to respond to retina-conditioned medium. CONCLUSIONS: Diffusible factors secreted by the neural retina acted synergistically with basolateral stimulation to regulate the structure and function of RPE tight junctions. Serum on the apical side of the RPE monolayer inhibited the ability of retinal factors to upregulate the tight junction barrier. PMID- 12556418 TI - Partial characterization of retina-derived cone neuroprotection in two culture models of photoreceptor degeneration. AB - PURPOSE: To define the nature and estimate the molecular weight range of soluble endogenous retinal trophic activities on cone photoreceptor survival in two models of cone degeneration. METHODS: Diffusible factors from dissociated retinal cell cultures of 8-day normal-sighted (C57BL/6J) mice were tested for cone survival-promoting activity by two approaches and by using two independent photoreceptor degeneration models. In the first approach, mouse retinal cells were cultured on semi-permeable membranes apposed to dissociated cultures of chick embryo retina. In the second approach, conditioned medium was collected from normal mouse retinal cultures and added to embryonic chicken retina cultures or to retinal explants obtained from 5-week retinal degeneration (rd1) mice. In some experiments, conditioned medium was heated or sequentially fractionated in dialysis tubing with molecular weight cutoffs of 8, 15, and 25 kDa. The number of chicken cones and viability were determined by using morphologic criteria, colorimetric assays, and labeling with antibodies raised against visinin. Mouse cones were counted by differential double immunolabeling with antibodies against rhodopsin (rods) and arrestin (rods and cones). RESULTS: . Coculturing with normal mouse retinal cells delayed cone loss in dispersed embryonic chicken retina, by a maximum of 50% relative to the control. Conditioned medium derived from normal mouse retinas also significantly delayed cone loss in chicken cone cultures by a maximum of 1300%, compared with the control, and 40% in rd1 mouse retinal explant cultures. The survival activity in conditioned medium was destroyed by heat denaturation, and was partially retained by dialysis with a molecular weight cutoff of 25 kDa in both models. CONCLUSIONS: These strategies have identified cone-survival-stimulating activities in normal mouse retina, capable of acting across species and enhancing both structural protection and viability. Such molecules may represent candidates for clinical treatment of inherited retinal degeneration. PMID- 12556420 TI - Role of vascular endothelial growth factor and placental growth factors during retinal vascular development and hyaloid regression. AB - PURPOSE: Vascular endothelial growth factor (VEGF)-A and placental growth factor (PlGF) are members of a large group of homologous peptides identified as the VEGF family. Although VEGF-A is known to act as a potent angiogenic peptide in the retina, the vasoactive function of PlGF in this tissue is less well defined. This study has sought to elucidate the expression patterns and modulatory role of these growth factors during retinal vascular development and hyaloid regression in the neonatal mouse. METHODS: C57BL6J mice were killed at postnatal days (P)1, P3, P5, P7, P9, and P11. The eyes were enucleated and processed for in situ hybridization and immunocytochemistry and the retinas extracted for total protein or RNA. Separate groups of neonatal mice were also injected intraperitoneally daily from P2 through P9 with either VEGF-neutralizing antibody, PlGF neutralizing antibody, isotype immunoglobulin (Ig)-G, or phosphate-buffered saline (PBS). The mice were then perfused with fluorescein isothiocyanate (FITC) dextran, and the eyes were subsequently embedded in paraffin wax or flat mounted. RESULTS: Quantitative (real-time) reverse transcription-polymerase chain reaction (RT-PCR) demonstrated similar expression patterns of VEGF-A and PlGF mRNA during neonatal retinal development, although the fluctuation between time periods was greater overall for VEGF-A. The localization of VEGF-A and PlGF in the retina, as revealed by in situ hybridization and immunohistochemistry, was also similar. Neutralization of VEGF-A caused a significant reduction in the hyaloid and retinal vasculature, whereas PlGF antibody treatment caused a marked persistence of the hyaloid without significantly affecting retinal vascular development. CONCLUSIONS: Although having similar expression patterns in the retina, these growth factors appear to have distinct modulatory influences during normal retinal vascular development and hyaloid regression. PMID- 12556419 TI - An RCS-like retinal dystrophy phenotype in mer knockout mice. AB - PURPOSE: To determine whether mice that are homozygous for a targeted disruption of the Mer receptor tyrosine kinase gene (mer(kd)) manifest a retinal dystrophy phenotype similar to RCS rats, which carry a mutation in the orthologous gene MERTK: METHODS: Eyes of mer(kd) and C57BL/6 wild-type (WT) mice were examined by light and electron microscopy, whole-eye rhodopsin measurement, and Ganzfeld electroretinography (ERG). RESULTS: The mer(kd) mice showed rapid, progressive degeneration of the photoreceptors (PRs). Features of the phenotype common to mer(kd) mice and RCS rats included the absence or near absence of phagosomes in the retinal pigment epithelium (RPE) at the peak of outer segment (OS) disc shedding, accumulation of debris and whorls of membranes at the RPE-OS interface, transient supernormal rhodopsin content and OS lengths, the presence of OS vacuoles beginning at early ages, and a relatively slow removal of pyknotic PR nuclei. Most PRs were missing, and OS debris was removed by approximately postnatal day (P)45. Scotopic ERG responses were lower than age-matched WT responses and declined with PR loss. Photopic responses were preserved better than scotopic responses, corresponding with preferential cone preservation as judged histologically. ERG amplitudes were usually unmeasurable beyond P40, although a small-amplitude scotopic threshold response (STR) could still be elicited at P253 in some mice when only scattered PR nuclei remained. CONCLUSIONS: Ablation of Mer function in mer(kd) mice results in a retinal phenotype almost identical with that of RCS rats. The similarity in phenotypes between the two rodent models suggests that an RPE phagocytic defect is a feature of all types of retinal degeneration caused by loss of function of Mer tyrosine kinase, perhaps including mutations in human MERTK. PMID- 12556421 TI - A morphometric study of light-induced damage in transgenic rat models of retinitis pigmentosa. AB - PURPOSE: To determine relative susceptibility to, and regional variation of, light-induced retinal damage in two rhodopsin-mutant rat models of retinitis pigmentosa, using slow- and fast-degenerating lines. METHODS: Transgenic S334ter (lines 4 and 9) and P23H (lines 2 and 3) rats were reared in dim cyclic light or darkness and then exposed to intense green light for 1 to 8 hours. Sections along the vertical meridian were collected for retinal morphology and photoreceptor morphometry 2 weeks later. Unexposed transgenic and normal Sprague-Dawley rats served as the control. Mean outer segment lengths and outer nuclear layer thicknesses were analyzed as a function of position along the vertical meridian and as averages across that vector. RESULTS: Rapidly degenerating S334ter-4 retinas, reared in dim cyclic light, exhibited no light-induced damage, whereas retinas in the other sublines sustained damage within a sensitive region in the superior hemisphere. Light-induced damage always involved loss of outer segment membrane and photoreceptors. In some cases, the retinal pigment epithelium and inner nuclear layer were also affected. Potentiation of light-induced damage by dark-rearing was increased by at least a factor of three, and in some sublines the sensitive region was enlarged to include the entire vertical meridian. CONCLUSIONS: A complex pattern of light-induced damage outcomes was identified in S334ter (sublines 4 and 9) and P23H (sublines 2 and 3) rats. The relative susceptibilities of each subline to damage by light were different, even within the same transgene, but consistent factors included a sensitive region in the superior hemisphere and potentiation by dark-rearing. PMID- 12556422 TI - Neuronal migration and glial remodeling in degenerating retinas of aged rats and in nonneovascular AMD. AB - PURPOSE: To demonstrate structural and immunocytochemical changes associated with light-induced degeneration in albino rat retinas and human AMD retinas. METHODS: Retinas from Wistar rats aged 3, 6, or 10 months were examined by immunocytochemistry, with antibodies to neuronal and glial markers. Results were compared with human nonneovascular AMD retinas. RESULTS: In aging rat retinas, many photoreceptors were lost in response to normal ambient light exposure. Photoreceptor loss was preceded by loss of RPE cells. Muller cells extended processes through gaps in Bruch's membrane, into the choroid. immunolabeling for gamma-aminobutyric acid (GABA), the glycine transporter Glyt-1, and the rod bipolar cell marker PKC revealed the presence of numerous neuronal somata and processes that appeared to have migrated into the choroidal region. Processes of presumptive ganglion cells remodeled and stratified in the choroid, where strong labeling for synaptic vesicle antigens was present. Myelination of retinal ganglion cell axons was also observed, especially in the peripheral retina. In AMD retinas, glial rearrangement and displacement of neurons suggestive of their migration were also observed. CONCLUSIONS: In response to loss of RPE and photoreceptor cells, adult retinal neurons migrate out of the retina along remodeled processes of Muller cells. The presence of synaptic vesicle antigens suggests the formation of new synapses between migrating neurons. The myelination is probably due to the ingress of Schwann cells from the sclera. The presence of some similar changes in human AMD retinas suggests that these findings are of broad significance for determining the likely events in transplantation of neurons in the human retina and elsewhere. PMID- 12556423 TI - Regional cone dysfunction in retinitis pigmentosa evaluated by flicker ERGs: relationship with perimetric sensitivity losses. AB - PURPOSE: To evaluate regional cone dysfunction in retinitis pigmentosa (RP) by recording focal electroretinograms (FERGs) from the central and paracentral retinal regions and to correlate the FERG with perimetric sensitivity losses. METHODS: Twenty-three typical patients with RP (age, 18-65 years; visual acuity, 20/100 to 20/20; kinetic visual field by size II/4e, 20-40 degrees ) and eight age-matched control subjects were evaluated. FERGs were recorded in response to either a central (eccentricity, 0-2.25 degrees ) or a paracentral annular (2.25-9 degrees ) field, presented on a light-adapting background. Fields' luminances (mean: 80 cd/m(2)) were sinusoidally modulated at different temporal frequencies (TFs; 10.3, 14, 21, 32, 41, and 52 Hz). Amplitude and phase of the responses' fundamental harmonic (1F) were measured. Perimetric sensitivity was measured by a visual field perimeter. For each patient, mean sensitivity losses were calculated for both the central (0-2.25 degrees ) and paracentral (2.25-9 degrees ) regions. RESULTS: On average, central and paracentral FERGs of patients with RP were reduced in amplitude (P 50% of its strains resistant. The effective control of these minor penicillin-resistant serotypes should be based on continuous surveillance of pneumococcal epidemiology. PMID- 12556430 TI - Susceptibility patterns of bacteria causing community-acquired respiratory infections in Spain: the SAUCE project. AB - Multicentre surveillance is essential in order to monitor the prevalence of certain resistance phenotypes and to identify rapidly the emergence of new ones. However, many surveillance studies are based either on a relatively small number of isolates from a single country, or on a large number of isolates from many different countries and so are not equally meaningful. Extensive national multicentre surveillance would provide a more reliable strategy for assessing the extent of antimicrobial resistance in individual countries. This article describes Spanish experience with the surveillance network SAUCE, and summarizes the main results on antimicrobial resistance in the three key bacterial pathogens involved in community-acquired respiratory tract infections in Spain: Streptococcus pneumoniae, Haemophilus influenzae and Streptococcus pyogenes. PMID- 12556431 TI - Antibiotic consumption and resistance selection in Streptococcus pneumoniae. AB - Selection of antibiotic-resistant Streptococcus pneumoniae is an inescapable consequence of antibiotic use. The correlation between antibiotic consumption and selection of resistant organisms can be shown at every ecological level: patient, community, region or country. In the case of multiple resistance, the intensity of antibiotic selection is increased. However, different antibiotics may exert different selective powers. Because of this, co-selection of macrolide and beta lactam resistance is an asymmetrical phenomenon: macrolides select more efficiently strains resistant to both macrolides and beta-lactams than aminopenicillins. The difference in rates of antibiotic resistance is also influenced by the local spread of susceptible or resistant clones; it is suggested that under mild antibiotic selection, the susceptible organisms that are more fit for host-to-host transmission could be favoured. Subsequent acquisition of resistance in these clones may rapidly increase the prevalence of resistance, and that may lead to an increase in the use of antibiotics. The reasons for antibiotic resistance are mainly the reasons explaining antibiotic consumption. A number of possible sociobiological determinants of antibiotic consumption can be identified: genetic factors in the human populations (for instance, involving different symptomatic types of infection), cultural factors and attitudes of patients towards antibiotics, sociological factors or public health factors, including incidence of other infectious diseases in the human population. Only 'excess' in the use of antibiotics should be controlled; for such a purpose, the concept, 'appropriate demand for antibiotics' (ADA) is proposed. PMID- 12556432 TI - Importance of dose and duration of beta-lactam therapy in nasopharyngeal colonization with resistant pneumococci. AB - Antibiotic use greatly influences the nasopharyngeal carriage of drug-resistant Streptococcus pneumoniae and is considered to be one of the most important risk factors for carriage of such organisms. Several studies have shown a reduction in the prevalence of resistant strains following reduced antibiotic consumption. This indicates that it may be possible to reduce resistance rates by changing prescribing patterns. Studies suggest that antibiotics present at low levels tend to select strains with low-level penicillin resistance, that intermediate antibiotic levels may pose the danger of selection of pneumococci with high-level penicillin resistance and that attainment of high levels of antibiotics may reduce the selective pressure for penicillin resistance. High-dose beta-lactam (amoxicillin) therapy has been shown to reduce the selection of resistant pneumococci in the nasopharynx. Likewise, short-course antibiotic therapy has reduced colonization by resistant strains. Finally, the effect of the antibiotic appears to differ according to the beta-lactam prescribed. Studies suggest that selection by cephalosporins occurs at higher frequencies than that by amoxicillin; this may be explained by the reduced activity of cephalosporins against penicillin-resistant S. pneumoniae. PMID- 12556433 TI - Nasopharyngeal carriage of Streptococcus pneumoniae: a review of the potential role of pharmacokinetics and pharmacodynamics. AB - Much antibiotic prescribing is based upon irrational practice rather than scientific principles. The advent and the better understanding of the pharmacokinetics and pharmacodynamics of antimicrobials are slowly leading to a change whereby antibiotics are prescribed with a better rationale. The fundamental understanding of the pharmacokinetics of the antimicrobial, for example how well it is absorbed, metabolized, distributed and eliminated from the body, should lead to an appreciation of the correct dosing interval. Pharmacodynamic factors, in particular, appreciation of the MIC of an antimicrobial for an infecting pathogen and whether time-independent or dependent killing occurs, will also have a profound influence upon how an antimicrobial is used. This review attempts to consider these features as they apply to nasopharyngeal carriage of Streptococcus pneumoniae. The reason for the dosing and dosing frequency of a number of agents is discussed and an attempt is made to point the way to more judicious antibiotic prescribing so as to maximize clinical efficacy yet minimize the possibility of resistance emerging during therapy. PMID- 12556434 TI - Pharmacodynamic and kinetic basis for the selection of pneumococcal resistance in the upper respiratory tract. AB - The oropharynx is both the reservoir where antibiotic-resistant Streptococcus pneumoniae strains are selected and the focus for further spread of these organisms. In order to select antibiotic-resistant organisms, the antibiotic must be able to eradicate the susceptible population. How an antibiotic acts on oropharyngeal flora is not fully understood, although data on salivary antibiotic concentrations could be useful for establishing some correlation. Many beta lactam antibiotics are very active against S. pneumoniae, and although their saliva concentrations are very low, there may be enough to eradicate the majority of beta-lactam-sensitive strains. Other antibiotics achieve salivary concentrations in the range 10-30% of serum concentrations (erythromycin, clindamycin, doxycycline and rifampicin), and are also able to eradicate the antibiotic-susceptible population. Antibiotics achieving higher salivary concentrations (>/=40% of the serum levels) are not very active against S. pneumoniae (ciprofloxacin) or are able to eradicate antibiotic-sensitive and intermediate strains (clarithromycin, azithromycin and telithromycin). Only antibiotics for which there are no highly resistant pneumococcal strains, for instance some beta-lactams administered at very high dose and for a short course, are associated with a lower risk of antibiotic resistance selection. To diminish the risk, the antibiotics should be dosed in order to obtain inhibitory quotients (maximal serum concentration/MIC ratios) >/= 4, which depends not only on the antibiotic concentration achieved but also on the lack of highly resistant organisms. PMID- 12556435 TI - Dynamics of nasopharyngeal colonization by potential respiratory pathogens. AB - Studies have shown that colonization of the nasopharynx by potential respiratory pathogens Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis is established early in childhood, although rates vary greatly according to locality, sampling frequency, individual and social factors. Factors influencing colonization and elimination are not as yet fully understood, but adhesion to mucosal receptors and immune responses are implicated in addition to bacterial properties and colonization resistance dynamics. Colonization in children and adults has been intensively studied in various localities. Potential pathogens are more likely to colonize the nasopharynx of children prone to recurrent otitis media, where impaired local immunity and repeated exposure to respiratory pathogens are additional risk factors. Adults with chronic respiratory tract disease also have higher carriage rates. The factors contributing to increased risk of carriage of potential respiratory pathogens, as well as to clinical infection and antimicrobial resistance, are summarized in this review. PMID- 12556436 TI - From ecological reservoir to disease: the nasopharynx, day-care centres and drug resistant clones of Streptococcus pneumoniae. AB - Several lines of epidemiological and microbiological studies point to the multiple and critical roles of the nasopharynx of children-particularly those of pre-school age and attending day-care centres (DCCs)-in the emergence and spread of drug-resistant Streptococcus pneumoniae (DRP). A systematic yearly surveillance of the nasopharyngeal flora of children attending DCCs has been carried out in Lisbon since 1996. Molecular typing of several hundred DRP isolates showed that the great majority of DRP were represented by a relatively few clonal types that were frequently carried by many children in geographically distant DCCs and over several years of surveillance. The same epidemic DRP clones were also frequent among pneumococci causing both paediatric and adult disease worldwide. Penicillin-resistant pneumococci carry sequences of heterologous origin in their pbp genes and also in the recently identified murM: a gene essential for expression of penicillin resistance and for the unique cell wall structure of penicillin-resistant pneumococci. Virtually all DRP express only a limited number (five or six) of the very large genetic repertoire (up to 90) of serotypes available for this bacterial species and the serotypes of drug resistant strains happens to be the same as the serotypes of drug-susceptible pneumococci that most frequently colonize pre-school age children. These observations strongly suggest that the nasopharynx of children is an important global ecological reservoir of DRP and may also play a critical role as the favoured anatomical site for the evolution of DRP. PMID- 12556437 TI - Beta-lactam activity against resistant pneumococcal strains is enhanced by the immune system. AB - Pandemic resistance in Streptococcus pneumoniae is compromising antibiotic activity. Antibiotics that act on the cell wall, such as beta-lactams, may have a combined effect with the immune system against S. pneumoniae, since both act on the bacterial envelope. This combined effect can be studied in vitro or in vivo with respect to bacterial killing, since lysis is the end-point of both beta lactams and the immune system. We review here the in vitro increase in the bactericidal activity of aminopenicillins by non-specific immunity (complement and polymorphonuclear leucocytes). Few data are available on the collaboration of specific immunity and beta-lactams. We also review the effect of the presence of specific antibodies on the in vivo T > MIC needed for the therapeutic efficacy of amoxicillin, and on blood bacterial clearance in animal models. The effect that immunity has on pharmacodynamic parameters, such as T > MIC, in non-human studies may be used as a tool to predict the effect of these pharmacodynamic variations in overcoming resistance and its selection, in the context of increasing the use of pneumococcal conjugated vaccines. PMID- 12556438 TI - Implications of antimicrobial resistance in the empirical treatment of community acquired respiratory tract infections: the case of macrolides. AB - Macrolide resistance among pneumococci is increasing worldwide and is associated with increasing macrolide use. Recent studies show that use of macrolides and azalides increases nasopharyngeal carriage of both macrolide-resistant and penicillin-resistant pneumococci. Carriage of a resistant pneumococcus may foster dissemination. The clinical relevance of in vitro resistance has been debated. However, recent data from a matched case-control study showed that 18 (24%) of 76 patients had breakthrough bacteraemia with an erythromycin-resistant pneumococcus while taking a macrolide, whereas none of the 136 matched controls with an erythromycin-susceptible pneumococcal bacteraemia was taking a macrolide (P = 0.0000001). Moreover, five (24%) of 21 patients bacteraemic with the low-level resistant M phenotype and none of the 40 matched controls were taking a macrolide (P = 0.00157). These data indicate that macrolide resistance due to both the efflux and the methylase mechanisms is clinically relevant. Furthermore, they favour guidelines for the empirical treatment of outpatients with community acquired pneumonia that recommend high-dose oral amoxicillin and reserve coverage of atypical pathogens for selected high-risk populations. PMID- 12556439 TI - New strategies to overcome antimicrobial resistance in Streptococcus pneumoniae with beta-lactam antibiotics. AB - The worldwide epidemic of antibiotic resistance in Streptococcus pneumoniae appears to advance inexorably and the measures taken to date to contain its progression have not been successful. Consideration should be given to the complex relationship between antibiotic consumption and resistance to the drug administered and also to other agents in the same and other antibiotic groups. We propose an integrated triple strategy that maximizes the use of the current antibiotic arsenal and is designed to curb the spread of resistance in S. pneumoniae. There are three main parts to this strategy: (i) reduction of prescribing, with particular emphasis on those drugs whose consumption has been shown to correlate strongly with resistance; (ii) development of new formulations or dosing schedules of those drugs whose pharmacodynamic parameters are better suited to cope with highly resistant strains; and (iii) encouragement of the use of antibiotics with the maximal capability of bacterial eradication. We believe such a strategy would reduce the spread of resistance both in the current ecological situation and in the post-pneumococcal vaccination era to come. Of all current antibiotics amoxicillin meets the above requirements, and seems to be the least ecologically disturbing oral antibiotic with regard to resistance in S. pneumoniae. PMID- 12556441 TI - Solid-state NMR reveals structural changes in phospholamban accompanying the functional regulation of Ca2+-ATPase. AB - Calcium transport across the sarcoplasmic reticulum of cardiac myocytes is regulated by a reversible inhibitory interaction between the Ca2+-ATPase and the small transmembrane protein phospholamban (PLB). A nullcysteine analogue of PLB, containing isotope labels in the transmembrane domain or cytoplasmic domain, was reconstituted into membranes in the absence and presence of the SERCA1 isoform of Ca2+-ATPase for structural investigation by cross-polarization magic-angle spinning (CP-MAS) NMR. PLB lowered the maximal hydrolytic activity of SERCA1 and its affinity for calcium in membrane preparations suitable for structural analysis by NMR. Novel backbone amide proton-deuterium exchange CP-MAS NMR experiments on the two PLB analogues co-reconstituted with SERCA1 indicated that labeled residues Leu42 and Leu44 were situated well within the membrane interior, whereas Pro21 and Ala24 lie exposed outside the membrane. Internuclear distance measurements on PLB using rotational resonance NMR indicated that the sequences Pro21-Ala24 and Leu42-Leu44 adopt an alpha-helical structure in pure lipid bilayers, which is unchanged in the presence of Ca2+-ATPase. By contrast, rotational echo double resonance (REDOR) NMR experiments revealed that the sequence Ala24-Gln26 switches from an alpha-helix in pure lipid membranes to a more extended structure in the presence of SERCA1, which may reflect local structural distortions which change the orientations of the transmembrane and cytoplasmic domains. These results suggest that Ca2+-ATPase has a long-range effect on the structure of PLB around residue 25, which promotes the functional association of the two proteins. PMID- 12556442 TI - Antithrombin III phenylalanines 122 and 121 contribute to its high affinity for heparin and its conformational activation. AB - The dissociation equilibrium constant for heparin binding to antithrombin III (ATIII) is a measure of the cofactor's binding to and activation of the proteinase inhibitor, and its salt dependence indicates that ionic and non-ionic interactions contribute approximately 40 and approximately 60% of the binding free energy, respectively. We now report that phenylalanines 121 and 122 (Phe-121 and Phe-122) together contribute 43% of the total binding free energy and 77% of the energy of non-ionic binding interactions. The large contribution of these hydrophobic residues to the binding energy is mediated not by direct interactions with heparin, but indirectly, through contacts between their phenyl rings and the non-polar stems of positively charged heparin binding residues, whose terminal amino and guanidinium groups are thereby organized to form extensive and specific ionic and non-ionic contacts with the pentasaccharide. Investigation of the kinetics of heparin binding demonstrated that Phe-122 is critical for promoting a normal rate of conformational change and stabilizing AT*H, the high affinity activated binary complex. Kinetic and structural considerations suggest that Phe 122 and Lys-114 act cooperatively through non-ionic interactions to promote P helix formation and ATIII binding to the pentasaccharide. In summary, although hydrophobic residues Phe-122 and Phe-121 make minimal contact with the pentasaccharide, they play a critical role in heparin binding and activation of antithrombin by coordinating the P-helix-mediated conformational change and organizing an extensive network of ionic and non-ionic interactions between positively charged heparin binding site residues and the cofactor. PMID- 12556443 TI - The C-terminal fragment of presenilin 2 triggers p53-mediated staurosporine induced apoptosis, a function independent of the presenilinase-derived N-terminal counterpart. AB - Mutations on presenilins are responsible for most of familial forms of Alzheimer's disease. These holoproteins undergo rapid maturation by presenilinase mainly in the endoplasmic reticulum, leading to the production of N- and C terminal fragments. We show first that overexpression of the presenilinase derived maturation product of presenilin 2 (CTF-PS2) increases Abeta recovery, the production of which is almost abolished by a caspase 3 inhibitor and increased by staurosporine. This and the observation that the apoptotic inducer staurosporine enhances CTF-PS2 degradation clearly link CTF-PS2 to apoptotic cascade effectors. This prompted us to analyze the putative ability of CTF-PS2 to modulate cell death. CTF-PS2 overexpression decreases cell viability and augments both caspase 3 activity and immunoreactivity. This is accompanied by lowered bcl2 like immunoreactivity and increased poly(ADP-ribose) polymerase cleavage and cytochrome c translocation into the cytosol. Interestingly, CTF-PS2-induced caspase 3 activation is prevented by pifithrin-alpha, a selective blocker of p53 transcriptional activity. On line with the latter data, CTF-PS2 drastically increases p53 immunoreactivity and transcriptional activity. Of most interest is our observation that CTF-PS2 expression also triggers increased caspase 3 activity and immunoreactivity in fibroblasts in which presenilins had been deleted. Therefore, CTF-PS2 could modulate cell death out of the NTF/CTF heterodimeric complex thought to correspond to the biologically functional entity. This is the first direct demonstration that CTF-PS2 could exhibit some of its functions in the absence of the presenilin 2 N-terminal fragment (NTF-PS2) counterpart derived from the presenilinase cleavage. PMID- 12556444 TI - Inhibition of glucose metabolism sensitizes tumor cells to death receptor triggered apoptosis through enhancement of death-inducing signaling complex formation and apical procaspase-8 processing. AB - Tumors display a high rate of glucose uptake and glycolysis. We investigated how inhibition of glucose metabolism could affect death receptor-mediated apoptosis in human tumor cells of diverse origin. We show that both substitution of glucose for pyruvate and treatment with 2-deoxyglucose enhanced apoptosis induced by tumor necrosis factor (TNF)-alpha, CD95 agonistic antibody, and TNF-related apoptosis-inducing ligand (TRAIL). Inhibition of glucose metabolism enhanced killing of myeloid leukemia U937, cervical carcinoma HeLa, and breast carcinoma MCF-7 cells upon death receptor ligation. Caspase activation, mitochondrial depolarization, and cytochrome c release were increased under these conditions. Glucose deprivation-mediated sensitization to apoptosis was prevented in MCF-7 cells overexpressing BCL-2. Interestingly, the human B-lymphoblastoid cell line SKW6.4, a prototype for mitochondria-independent death receptor-induced apoptosis, was also sensitized to anti-CD95 and TRAIL-induced apoptosis under glucose-free conditions. Changes in c-FLIP(L) and cFLIPs levels were observed in some but not all the cell lines studied following glucose deprivation. Glucose deprivation enhanced death receptor-triggered formation of death-inducing signaling complex and early processing of procaspase-8. Altogether, these results suggest that the glycolytic pathway may be an important target for therapeutic intervention to sensitize tumor cells to selectively toxic soluble death ligands or death ligand-expressing cells of the immune system by facilitating the activation of initiator caspase-8. PMID- 12556445 TI - A molecular docking strategy identifies Eosin B as a non-active site inhibitor of protozoal bifunctional thymidylate synthase-dihydrofolate reductase. AB - Protozoal parasites are unusual in that their thymidylate synthase (TS) and dihydrofolate reductase (DHFR) enzymes exist on a single polypeptide. In an effort to probe the possibility of substrate channeling between the TS and DHFR active sites and to identify inhibitors specific for bifunctional TS-DHFR, we used molecular docking to screen for inhibitors targeting the shallow groove connecting the two active sites. Eosin B is a 100 microm non-active site inhibitor of Leishmania major TS-DHFR identified by molecular docking. Eosin B slows both the TS and DHFR reaction rates. When Arg-283, a key residue to which eosin B is predicted to bind, is mutated to glutamate, however, eosin B only minimally inhibits the TS-DHFR reaction. Additionally, eosin B was found to be a 180 microm inhibitor of Toxoplasma gondii in both biochemical and cell culture assays. PMID- 12556446 TI - Amphetamine-induced dopamine efflux. A voltage-sensitive and intracellular Na+ dependent mechanism. AB - Amphetamine (AMPH) elicits its behavioral effects by acting on the dopamine (DA) transporter (DAT) to induce DA overflow into the synaptic cleft. Facilitated exchange diffusion is the classical model used to describe AMPH-induced DA efflux. This model hypothesizes that AMPH-induced DA efflux is mediated by DAT and results from the transport of AMPH into the cell followed by a counter movement of DA out to the extracellular compartment. To further characterize the action of AMPH, we used the patch clamp technique in the whole-cell configuration combined with amperometry on human embryonic kidney HEK-293 cells stably transfected with the human DAT (DAT cells). In DAT cells, AMPH-induced DAT mediated currents were blocked by cocaine. We demonstrate that DA efflux mediated by DAT is voltage-dependent, electrogenic, and dependent on intracellular Na(+) concentration in the recording electrode. Intracellular Na(+) fluorescence, as measured by confocal microscopy using a Na(+)-sensitive dye, was enhanced by AMPH application. Furthermore, the ability of AMPH to induce DA efflux was regulated by intracellular Na(+) concentration and correlated with the size of the DAT mediated, AMPH-induced ion flux across the plasma membrane. In the absence of intracellular Na(+) but the presence of high intracellular Cl(-), AMPH-induced inward currents elicited DA efflux proportionally to their dimension and duration. Thus, we propose that AMPH-induced DA efflux depends on two correlated transporter processes. First, AMPH binds to the DAT and is transported, thereby causing an inward current. Second, because of this AMPH-induced inward current, Na(+) becomes more available intracellularly to the DAT, thereby enhancing DAT mediated reverse transport of DA. PMID- 12556447 TI - An extended LXXLL motif sequence determines the nuclear receptor binding specificity of TRAP220. AB - The interaction of coactivators with the ligand-binding domain of nuclear receptors (NRs) is mediated by amphipathic alpha-helices containing the signature motif LXXLL. TRAP220 contains two LXXLL motifs (LXM1 and LXM2) that are required for its interaction with NRs. Here we show that the nuclear receptor interaction domain (NID) of TRAP220 interacts weakly with Class I NRs. In contrast, SRC1 NID binds strongly to both Class I and Class II NRs. Interaction assays using nine amino acid LXXLL core motifs derived from SRC1 and TRAP220 revealed no discriminatory NR binding preferences. However, an extended LXM1 sequence containing amino acids -4 to +9, (where the first conserved leucine is +1) showed selective binding to thyroid hormone receptor and reduced binding to estrogen receptor. Replacement of either TRAP220 LXXLL motif with the corresponding 13 amino acids of SRC1 LXM2 strongly enhanced the interaction of the TRAP220 NID with the estrogen receptor. Mutational analysis revealed combinatorial effects of the LXM1 core and flanking sequences in the determination of the NR binding specificity of the TRAP220 NID. In contrast, a mutation that increased the spacing between TRAP220 LXM1 and LXM2 had little effect on the binding properties of this domain. Thus, a 13-amino acid sequence comprising an extended LXXLL motif acts as the key determinant of the NR binding specificity of TRAP220. Finally, we show that the NR binding specificity of full-length TRAP220 can be altered by swapping extended LXM sequences. PMID- 12556448 TI - Role of caspases, Bid, and p53 in the apoptotic response triggered by histone deacetylase inhibitors trichostatin-A (TSA) and suberoylanilide hydroxamic acid (SAHA). AB - Histone deacetylase activity is potently inhibited by hydroaximc acid derivatives such as suberoylanilide hydroxamic acid (SAHA) and trichostatin-A (TSA). These inhibitors specifically induce differentiation/apoptosis of transformed cells in vitro and suppress tumor growth in vivo. Because of its low toxicity, SAHA is currently evaluated in clinical trials for the treatment of cancer. SAHA and TSA induce apoptosis, which is characterized by mitochondrial stress, but so far, the critical elements of this apoptotic program remain poorly defined. To characterize in more detail this apoptotic program, we used human cell lines containing alterations in important elements of apoptotic response such as: p53, Bcl-2, caspase-9, and caspase-3. We demonstrate that caspase-9 is critical for apoptosis induced by SAHA and TSA and that efficient proteolytic activation of caspase-2, caspase-8, and caspase-7 strictly depends on caspase-9. Bcl-2 efficiently antagonizes cytochrome c release and apoptosis in response to both histone deacetylase inhibitors. We provide evidences that translocation into the mitochondria of the Bcl-2 family member Bid depends on caspase-9 and that this translocation is a late event during TSA-induced apoptosis. We also demonstrate that the susceptibility to TSA- and SAHA-induced cell death is regulated by p53. PMID- 12556449 TI - Modulation, via protein-protein interactions, of glyceraldehyde-3-phosphate dehydrogenase activity through redox phosphoribulokinase regulation. AB - The activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) embedded in the phosphoribulokinase (PRK).GAPDH.CP12 complex was increased 2-3-fold by reducing agents. This occurred by interaction with PRK as the cysteinyl sulfhydryls (4 SH/subunit) of GAPDH within the complex were unchanged whatever the redox state of the complex. But isolated GAPDH was not activated. Alkylation plus mass spectrometry also showed that PRK had one disulfide bridge and three SH groups per monomer in the active oxidized complex. Reduction disrupted this disulfide bridge to give 2 more SH groups and a much more active enzyme. We assessed the kinetics and dynamics of the interactions between PRK and GAPDH/CP12 using biosensors to measure complex formation in real time. The apparent equilibrium binding constant for GAPDH/CP12 and PRK was 14 +/- 1.6 nm for oxidized PRK and 62 +/- 10 nm for reduced PRK. These interactions were neither pH- nor temperature dependent. Thus, the dynamics of PRK.GAPDH.CP12 complex formation and GAPDH activity are modulated by the redox state of PRK. PMID- 12556450 TI - Identification of a functionally important conformation-sensitive region of the secretory Na+-K+-2Cl- cotransporter (NKCC1). AB - The secretory Na(+)-K(+)-2Cl(-) cotransporter (NKCC1) is a member of a small gene family of electroneutral salt transporters that play essential roles in salt and water homeostasis in many mammalian tissues. We have identified a highly conserved residue (Ala-483) in the sixth membrane-spanning segment of rat NKCC1 that when mutated to cysteine renders the transporter sensitive to inhibition by the sulfhydryl reagents 2-aminoethyl methanethiosulfonate (MTSEA) and 2 (trimethylammonium)ethyl methanethiosulfonate (MTSET). The mutation of Ala-483 to cysteine (A483C) results in little or no change in the affinities of NKCC1 for substrate ions but produces a 6-fold increase in sensitivity to the inhibitor bumetanide, suggesting a specific modification of the bumetanide binding site. When residues surrounding Ala-483 were mutated to cysteine, only I484C was sensitive to inhibition by MTSEA and MTSET. Surprisingly I484C showed increased transport activity in the presence of low concentrations of mercury (1-10 microm), whereas A483C showed inhibition. The inhibition of A483C by MTSEA was unaffected by the presence or absence of sodium and potassium but required the presence of extracellular chloride. Taken together, our results indicate that Ala 483 lies at or near an important functional site of NKCC1 and that the exposure of this site to the extracellular medium is dependent on the conformation of the transporter. Specifically, our results indicate that the cysteine introduced at residue 483 is only available for interaction with MTSEA when chloride is bound to NKCC1 at the extracellular surface. PMID- 12556451 TI - The LIM protein FHL3 binds basic Kruppel-like factor/Kruppel-like factor 3 and its co-repressor C-terminal-binding protein 2. AB - The ability of DNA-binding transcription factors to recruit specific cofactors is central to the mechanism by which they regulate gene expression. BKLF/KLF3, a member of the Kruppel-like factor family of zinc finger proteins, is a potent transcriptional repressor that recruits a CtBP co-repressor. We show here that BKLF also recruits the four and a half LIM domain protein FHL3. Different but closely linked regions of BKLF mediate contact with CtBP2 and FHL3. We present evidence that CtBP2 also interacts with FHL3 and demonstrate that the three proteins co-elute in gel filtration experiments. CtBP and FHL proteins have been implicated in both nuclear and cytoplasmic functions, but expression of BKLF promotes the nuclear accumulation of both FHL3 and CtBP2. FHL proteins have been shown to act predominantly as co-activators of transcription. However, we find FHL3 can repress transcription. We suggest that LIM proteins like FHL3 are important in assembling specific repression or activation complexes, depending on conditions such as cofactor availability and promoter context. PMID- 12556452 TI - Costameres: the Achilles' heel of Herculean muscle. PMID- 12556453 TI - Integrins: redundant or important players in skeletal muscle? PMID- 12556454 TI - The basement membrane/basal lamina of skeletal muscle. PMID- 12556455 TI - Dystrophin-glycoprotein complex: post-translational processing and dystroglycan function. PMID- 12556456 TI - Skeletal muscle basement membrane-sarcolemma-cytoskeleton interaction minireview series. PMID- 12556457 TI - Proteolytic processing and oligomerization of bacteriophage-derived endosialidases. AB - Bacteriophages infecting the neuroinvasive pathogen Escherichia coli K1 require an endosialidase to penetrate the polysialic acid capsule of the host. Sequence information is available for the endosialidases endoNE, endoNF, and endoN63D of the K1-specific phages phi K1E, phi K1F, and 63D, respectively. The cloned sequences share a highly conserved catalytic domain but differ in the length of the N- and C-terminal parts. Although the expression of active recombinant enzyme succeeded in the case of endoNE, it failed for endoNF. Protein alignments of all three endosialidase sequences gave rise to the assumption that inactivity of the cloned endoNF is caused by a C-terminal truncation. By reinvestigation of the respective gene locus in the phi K1F genome, we identified an extended open reading frame of 3195 bp, encoding a 119-kDa protein. Full-length endoNF contains the C-terminal domain conserved in all endosialidases, which may act as an intramolecular chaperone. Comparative studies carried out with endoNE and endoNF demonstrate that endosialidases are proteolytically processed, releasing the C terminal domain. Using a mutational approach in combination with protein analytical techniques we demonstrate that (i) the C-terminal domain is a common feature of endosialidases and other tail fiber proteins; (ii) the integrity of the C-terminal domain and its presence in the nascent protein are crucial for the formation of active enzymes; (iii) proteolytic processing is not essential for enzymatic activity; and (iv) functional folding is a prerequisite for trimerization of endoNF. PMID- 12556458 TI - gamma-Secretase cleavage site specificity differs for intracellular and secretory amyloid beta. AB - The final step in A beta generation is the cleavage of the C-terminal 99 amino acid residues of the amyloid precursor protein by gamma-secretase. gamma Secretase activity is closely linked to the multi-transmembrane-spanning proteins presenilin 1 and presenilin 2. To elucidate whether the cleavage site specificities of gamma-secretase leading to the formation of secreted and intracellular A beta are identical, we made use of point mutations close to the gamma-cleavage site, known to have a dramatic effect on the 42/40 ratio of secreted A beta. We found that the selected point mutations only marginally influenced the 42/40 ratio of intracellular A beta, suggesting differences in the gamma-secretase cleavage site specificity for the generation of secreted and intracellular A beta. The analysis of the subcellular compartments involved in the generation of intracellular A beta revealed that A beta is not generated in the early secretory pathway in the human SH-SY5Y neuroblastoma cell line. In this study we identified late Golgi compartments to be involved in the generation of intracellular A beta. Moreover, we demonstrate that the presence of processed PS1 is not sufficient to obtain gamma-secretase processing of the truncated amyloid precursor protein construct C99, proposing the existence of an additional factor downstream of the endoplasmic reticulum and early Golgi required for the formation of an active gamma-secretase complex. PMID- 12556459 TI - Prostaglandin E2 protects gastric mucosal cells from apoptosis via EP2 and EP4 receptor activation. AB - Prostaglandin E(2) (PGE(2)) has a strong protective effect on the gastric mucosa in vivo; however, the molecular mechanism of a direct cytoprotective effect of PGE(2) on gastric mucosal cells has yet to be elucidated. Although we reported previously that PGE(2) inhibited gastric irritant-induced apoptotic DNA fragmentation in primary cultures of guinea pig gastric mucosal cells, we show here that PGE(2) inhibits the ethanol-dependent release of cytochrome c from mitochondria. Of the four main subtypes of PGE(2) receptors, we also demonstrated, using subtype-specific agonists, that EP(2) and EP(4) receptors are involved in the PGE(2)-mediated protection of gastric mucosal cells from ethanol induced apoptosis. Activation of EP(2) and EP(4) receptors is coupled with an increase in cAMP, for which a cAMP analogue was found here to inhibit the ethanol induced apoptosis. The increase in cAMP is known to activate both protein kinase A (PKA) and phosphatidylinositol 3-kinase pathways. An inhibitor of PKA but not of phosphatidylinositol 3-kinase blocked the PGE(2)-mediated protection of cells from ethanol-induced apoptosis, suggesting that a PKA pathway is mainly responsible for the PGE(2)-mediated inhibition of apoptosis. Based on these results, we considered that PGE(2) inhibited gastric irritant-induced apoptosis in gastric mucosal cells via induction of an increase in cAMP and activation of PKA, and that this effect was involved in the PGE(2)-mediated protection of the gastric mucosa from gastric irritants in vivo. PMID- 12556460 TI - Membrane binding mechanisms of the PX domains of NADPH oxidase p40phox and p47phox. AB - Phox (PX) domains are phosphoinositide (PI)-binding domains with broad PI specificity. Two cytosolic components of NADPH oxidase, p40(phox) and p47(phox), contain PX domains. The PX domain of p40(phox) specifically binds phosphatidylinositol 3-phosphate, whereas the PX domain of p47(phox) has two lipid binding sites, one specific for phosphatidylinositol 3,4-bisphosphate and the other with affinity for phosphatidic acid or phosphatidylserine. To delineate the mechanisms by which these PX domains interact with PI-containing membranes, we measured the membrane binding of these domains and respective mutants by surface plasmon resonance and monolayer techniques and also calculated the electrostatic potentials of the domains as a function of PI binding. Results indicate that membrane binding of both PX domains is initiated by nonspecific electrostatic interactions, which is followed by the membrane penetration of hydrophobic residues. The membrane penetration of the p40(phox) PX domain is induced by phosphatidylinositol 3-phosphate, whereas that of the p47(phox) PX domain is triggered by both phosphatidylinositol 3,4-bisphosphate and phosphatidic acid (or phosphatidylserine). Studies of enhanced green fluorescent protein-fused PX domains in HEK293 cells indicate that this specific membrane penetration is also important for subcellular localization of the two PX domains. Further studies on the full-length p40(phox) and p47(phox) proteins showed that an intramolecular interaction between the C-terminal Src homology 3 domain and the PX domain prevents the nonspecific monolayer penetration of p47(phox), whereas such an interaction is absent in p40(phox). PMID- 12556461 TI - Purification and characterization of PrbA, a new esterase from Enterobacter cloacae hydrolyzing the esters of 4-hydroxybenzoic acid (parabens). AB - The esterase PrbA from Enterobacter cloacae strain EM has previously been shown to confer additional resistance to the esters of 4-hydroxybenzoic acid (parabens) to two species of Enterobacter. The PrbA protein has been purified from E. cloacae strain EM using a three-step protocol resulting in a 60-fold increase in specific activity. The molecular mass of the mature enzyme was determined to be 54,619 +/- 1 Da by mass spectrometry. It is highly active against a series of parabens with alkyl groups ranging from methyl to butyl, with K(m) and V(max) values ranging from 0.45 to 0.88 mM and 0.031 to 0.15 mM/min, respectively. The K(m) and V(max) values for p-nitrophenyl acetate were 3.7 mM and 0.051 mM/min. PrbA hydrolyzed a variety of structurally analogous compounds, with activities larger than 20% relative to propyl paraben for methyl 3-hydroxybenzoate, methyl 4 aminobenzoate, or methyl vanillate. The enzyme showed optimum activity at 31 degrees C and at pH 7.0. PrbA was able to transesterify parabens with alcohols of increasing chain length from methanol to n-butanol, achieving 64% transesterification of 0.5 mm propyl paraben with 5% methanol within 2 h. PrbA was inhibited by 1-chloro-3-tosylamido-4-phenyl-2-butanone and 1-chloro-3 tosylamido-7-amino-2-heptanone (TLCK), with K(i) values of 0.29 and 0.20 mM, respectively, and was irreversibly inhibited by Diisopropyl fluorophosphate (DFP) or diethyl pyrocarbonate. The stoichiometry of addition of DFP to the enzyme was 1:1 and only 1 TLCK molecule was found in TLCK-modified enzyme, as measured by mass spectrometry. Analysis of the tryptic digest of the DFP-modified PrbA demonstrated that the addition of a DFP molecule occurred at Ser-189, indicating the location of the active serine. PMID- 12556462 TI - Memoirs of a biochemical hod carrier. PMID- 12556463 TI - Using studies on tryptophan metabolism to answer basic biological questions. PMID- 12556464 TI - Musings. PMID- 12556465 TI - Determinants of the in vivo folding of the prion protein. A bipartite function of helix 1 in folding and aggregation. AB - Misfolding of the mammalian prion protein (PrP) is implicated in the pathogenesis of prion diseases. We analyzed wild type PrP in comparison with different PrP mutants and identified determinants of the in vivo folding pathway of PrP. The complete N terminus of PrP including the putative transmembrane domain and the first beta-strand could be deleted without interfering with PrP maturation. Helix 1, however, turned out to be a major determinant of PrP folding. Disruption of helix 1 prevented attachment of the glycosylphosphatidylinositol (GPI) anchor and the formation of complex N-linked glycans; instead, a high mannose PrP glycoform was secreted into the cell culture supernatant. In the absence of a C-terminal membrane anchor, however, helix 1 induced the formation of unglycosylated and partially protease-resistant PrP aggregates. Moreover, we could show that the C terminal GPI anchor signal sequence, independent of its role in GPI anchor attachment, mediates core glycosylation of nascent PrP. Interestingly, conversion of high mannose glycans to complex type glycans only occurred when PrP was membrane-anchored. Our study indicates a bipartite function of helix 1 in the maturation and aggregation of PrP and emphasizes a critical role of a membrane anchor in the formation of complex glycosylated PrP. PMID- 12556467 TI - Glutathione-thiyl radical scavenging and transferase properties of human glutaredoxin (thioltransferase). Potential role in redox signal transduction. AB - Glutaredoxin (GRx, thioltransferase) is implicated in cellular redox regulation, and it is known for specific and efficient catalysis of reduction of protein-S-S glutathione-mixed disulfides (protein-SSG) because of its remarkably low thiol pK(a) ( approximately 3.5) and its ability to stabilize a catalytic S glutathionyl intermediate (GRx-SSG). These unique properties suggested that GRx might also react with glutathione-thiyl radicals (GS(.)) and stabilize a disulfide anion radical intermediate (GRx-SSG), thereby facilitating the conversion of GS(.) to GSSG or transfer of GS(.) to form protein-SSG. We found that GRx catalyzes GSSG formation in the presence of GS-thiyl radical generating systems (Fe(2+)/ADP/H(2)O(2) + GSH or horseradish peroxidase/H(2)O(2) + GSH). Catalysis is dependent on O(2) and results in concomitant superoxide formation, and it is distinguished from glutathione peroxidase-like activity. With the horseradish peroxidase system and [(35)S]GSH, GRx enhanced the rate of GS radiolabel incorporation into GAPDH. GRx also enhanced the rate of S glutathionylation of glyceraldehyde-3-phosphate dehydrogenase with GSSG or S nitrosoglutathione, but these glutathionyl donors were much less efficient. Both actin and protein-tyrosine phosphatase-1B were superior substrates for GRx facilitated S-glutathionylation with GS-radical. These studies characterize GRx as a versatile catalyst, facilitating GS-radical scavenging and S glutathionylation of redox signal mediators, consistent with a critical role in cellular regulation. PMID- 12556466 TI - Egr1 promotes growth and survival of prostate cancer cells. Identification of novel Egr1 target genes. AB - In the majority of aggressive tumorigenic prostate cancer cells, the transcription factor Egr1 is overexpressed. We provide new insights of Egr1 involvement in proliferation and survival of TRAMP C2 prostate cancer cells by the identification of several new target genes controlling growth, cell cycle progression, and apoptosis such as cyclin D2, P19ink4d, and Fas. Egr1 regulation of these genes, identified by Affymetrix microarray, was confirmed by real-time PCR, immunoblot, and chromatin immunoprecipitation assays. Furthermore we also showed that Egr1 is responsible for cyclin D2 overexpression in tumorigenic DU145 human prostate cells. The regulation of these genes by Egr1 was demonstrated using Egr1 antisense oligonucleotides that further implicated Egr1 in resistance to apoptotic signals. One mechanism was illustrated by the ability of Egr1 to inhibit CD95 (Fas/Apo) expression, leading to insensitivity to FasL. The results provide a mechanistic basis for the oncogenic role of Egr1 in TRAMP C2 prostate cancer cells. PMID- 12556468 TI - Homotetrameric structure of the SNAP-23 N-terminal coiled-coil domain. AB - SNARE proteins mediate intracellular membrane fusion by forming a coiled-coil complex to merge opposing membranes. A "fusion-active" neuronal SNARE complex is a parallel four-helix bundle containing two coiled-coil domains from SNAP-25 and one coiled-coil domain each from syntaxin-1a and VAMP-2. "Prefusion" assembly intermediate complexes can also form from these SNAREs. We studied the N-terminal coiled-coil domain of SNAP-23 (SNAP-23N), a non-neuronal homologue of SNAP-25, and its interaction with other coiled-coil domains. SNAP-23N can assemble spontaneously with the coiled-coil domains from SNAP-23C, syntaxin-4, and VAMP-3 to form a heterotetrameric complex. Unexpectedly, pure SNAP-23N crystallizes as a coiled-coil homotetrameric complex. The four helices have a parallel orientation and are symmetrical about the long axis. The complex is stabilized through the interaction of conserved hydrophobic residues comprising the a and d positions of the coiled-coil heptad repeats. In addition, a central, highly conserved glutamine residue (Gln-48) is buried within the interface by hydrogen bonding between glutamine side chains derived from adjacent subunits and to solvent molecules. A comparison of the SNAP-23N structure to other SNARE complex structures reveals how a simple coiled-coil motif can form diverse SNARE complexes. PMID- 12556469 TI - Structural basis of calcification inhibition by alpha 2-HS glycoprotein/fetuin-A. Formation of colloidal calciprotein particles. AB - Genetic evidence from mutant mice suggests that alpha(2)-HS glycoprotein/fetuin-A (Ahsg) is a systemic inhibitor of precipitation of basic calcium phosphate preventing unwanted calcification. Using electron microscopy and dynamic light scattering, we demonstrate that precipitation inhibition by Ahsg is caused by the transient formation of soluble, colloidal spheres, containing Ahsg, calcium, and phosphate. These "calciprotein particles" of 30-150 nm in diameter are initially amorphous and soluble but turn progressively more crystalline and insoluble in a time- and temperature-dependent fashion. Solubilization in Ahsg-containing calciprotein particles provides a novel conceptual framework to explain how insoluble calcium precipitates may be transported and removed in the bodies of mammals. Mutational analysis showed that the basic calcium phosphate precipitation inhibition activity resides in the amino-terminal cystatin-like domain D1 of Ahsg. A structure-function analysis of wild type and mutant forms of cystatin-like domains from Ahsg, full-length fetuin-B, histidine-rich glycoprotein, and kininogen demonstrated that Ahsg domain D1 is most efficient in inhibiting basic calcium phosphate precipitation. The computer-modeled domain structures suggest that a dense array of acidic residues on an extended beta sheet of the cystatin-like domain Ahsg-D1 mediates efficient inhibition. PMID- 12556470 TI - A novel erythrocyte binding antigen-175 paralogue from Plasmodium falciparum defines a new trypsin-resistant receptor on human erythrocytes. AB - The recognition and invasion of human erythrocytes by the most lethal malaria parasite Plasmodium falciparum is dependent on multiple ligand-receptor interactions. Members of the erythrocyte binding-like (ebl) family, including the erythrocyte binding antigen-175 (EBA-175), are responsible for high affinity binding to glycoproteins on the surface of the erythrocyte. Here we describe a paralogue of EBA-175 and show that this protein (EBA-181/JESEBL) binds in a sialic acid-dependent manner to erythrocytes. EBA-181 is expressed at the same time as EBA-175 and co-localizes with this protein in the microneme organelles of asexual stage parasites. The receptor binding specificity of EBA-181 to erythrocytes differs from other members of the ebl family and is trypsin resistant and chymotrypsin-sensitive. Furthermore, using glycophorin B-deficient erythrocytes we show that binding of EBA-181 is not dependent on this sialoglycoprotein. The level of expression of EBA-181 differs among parasite lines, and the importance of this ligand for invasion appears to be strain dependent as the EBA-181 gene can be disrupted in W2mef parasites, without affecting the invasion phenotype, but cannot be targeted in 3D7 parasites. PMID- 12556471 TI - Negative regulation of T cell activation by placental protein 14 is mediated by the tyrosine phosphatase receptor CD45. AB - CD45 is the major protein tyrosine phosphatase receptor on T cell surfaces that functions as both a positive and a negative regulator of T cell receptor (TCR) signaling. Although CD45 is required for the activation of TCR-associated Src family kinases, it also dephosphorylates phosphoproteins involved in the TCR signaling cascade. This study links CD45 to the inhibitory activity of placental protein 14 (PP14), a major soluble protein of pregnancy that is now known to be a direct modulator of T cells and to function by desensitizing TCR signaling. PP14 and CD45 co-capped with each other, pointing to a physical linkage between the two. Interestingly, however, the binding of PP14 to T cell surfaces was not restricted to CD45 alone, with evidence showing that PP14 binds to other surface molecules in a carbohydrate-dependent fashion. Notwithstanding the broader molecular binding potential of PP14, its interaction with CD45 appeared to have special functional significance. Using transfected derivatives of the HPB. ALL mutant T cell line that differ in CD45 expression, we established that the inhibitory effects of PP14 are dependent upon the expression of intact CD45 on T cell surfaces. Based upon these findings, we propose a new immunoregulatory model for PP14, wherein one of its surface molecular targets, CD45, mediates its T cell inhibitory activity, accounting for the intriguing capacity of PP14 to elevate TCR activation thresholds and thereby down-regulate T cell activation. PMID- 12556472 TI - The GABAA receptor alpha 1 subunit Pro174-Asp191 segment is involved in GABA binding and channel gating. AB - The GABA-binding site undergoes structural rearrangements during the transition from agonist binding to channel opening. To define possible roles of the GABA(A) receptor alpha(1) subunit Pro(174)-Asp(191) segment in these processes, we used the substituted cysteine accessibility method to characterize this region. Each residue was individually mutated to cysteine, expressed with wild-type beta(2) subunits in Xenopus laevis oocytes, and examined using two-electrode voltage clamp. Most mutations did not alter GABA EC(50) values. The D183C mutation produced a 7-fold reduction in GABA sensitivity. There were no significant changes in the K(I) values for the competitive antagonist, SR-95531. N Biotinylaminoethyl methanethiosulfonate modified P174C-, R176C-, S177C-, V178C-, V180C-, A181C-, D183C-, R186C- and N188C-containing receptors. The pattern of accessibility suggests that this protein segment is aqueous-exposed and adopts a random coil conformation. Both GABA and SR-95531 slowed covalent modification of V178C, V180C, and D183C, indicating that these residues may line the GABA-binding site. Further, pentobarbital-induced channel activation accelerated modification of V180C and A181C and slowed the modification of R186C, suggesting that this region of the alpha(1) subunit may act as a dynamic element during channel-gating transitions. PMID- 12556473 TI - Identification of the F1-binding surface on the delta-subunit of ATP synthase. AB - The stator function in ATP synthase was studied by a combined mutagenesis and fluorescence approach. Specifically, binding of delta-subunit to delta-depleted F(1) was studied. A plausible binding surface on delta-subunit was identified from conservation of amino acid sequence and the high resolution NMR structure. Specific mutations aimed at modulating binding were introduced onto this surface. Affinity of binding of wild-type and mutant delta-subunits to delta-depleted F(1) was determined quantitatively using the fluorescence signals of natural delta-Trp 28, inserted delta-Trp-11, or inserted delta-Trp-79. The results demonstrate that helices 1 and 5 in the N-terminal domain of the delta-subunit provide the F(1) binding surface of delta. Unexpectedly, mutations that impaired binding between F(1) and delta were found to not necessarily impair ATP synthase activity. Further investigation revealed that inclusion of the soluble cytoplasmic domain of the b subunit substantially enhanced affinity of binding of delta-subunit to F(1). The new data show that the stator is "overengineered" to resist rotor torque during catalysis. PMID- 12556474 TI - The KH-domain protein alpha CP has a direct role in mRNA stabilization independent of its cognate binding site. AB - Previous studies suggest that high-level stability of a subset of mammalian mRNAs is linked to a C-rich motif in the 3' untranslated region (3'UTR). High-level expression of human alpha-globin mRNA (h alpha-globin mRNA) in erythroid cells has been specifically attributed to formation of an RNA-protein complex comprised of a 3'UTR C-rich motif and an associated 39-kDa poly(C) binding protein, alpha CP. Documentation of this RNA-protein alpha-complex has been limited to in vitro binding studies, and its impact has been monitored by alterations in steady-state mRNA. Here we demonstrate that alpha CP is stably bound to h alpha-globin mRNA in vivo, that alpha-complex assembly on the h alpha-globin mRNA is restricted to the 3'UTR C-rich motif, and that alpha-complex assembly extends the physical half life of h alpha-globin mRNA selectively in erythroid cells. Significantly, these studies also reveal that an artificially tethered alpha CP has the same mRNA stabilizing activity as the native alpha-complex. These data demonstrate a unique contribution of the alpha-complex to h alpha-globin mRNA stability and support a model in which the sole function of the C-rich motif is to selectively tether alpha CP to a subset of mRNAs. Once bound, alpha CP appears to be fully sufficient to trigger downstream events in the stabilization pathway. PMID- 12556475 TI - Effect of the pheromone-responsive G(alpha) and phosphatase proteins of Saccharomyces cerevisiae on the subcellular localization of the Fus3 mitogen activated protein kinase. AB - The mating-specific G(alpha) protein of Saccharomyces cerevisiae, Gpa1, stimulates adaptation to pheromone by a mechanism independent of G(beta gamma) sequestration. Genetic evidence suggests that Gpa1 targets the Fus3 mitogen activated protein kinase, and it has recently been shown that the two proteins interact in cells responding to pheromone. To test the possibility that Gpa1 downregulates the mating signal by affecting the localization of Fus3, we created a Fus3-green fluorescent protein (GFP) fusion protein. In vegetative cells, Fus3 GFP was found in both the cytoplasm and the nucleus. Pheromone stimulated a measurable increase in the ratio of nuclear to cytoplasmic Fus3-GFP. In contrast, the relative level of nuclear Fus3-GFP decreased as cells recovered from pheromone arrest and did not increase when cells adapted to chronic stimulus were challenged again. Accumulation of Fus3-GFP in the nuclei of stimulated cells was also inhibited by overexpression of either wild-type Gpa1, the E364K hyperadaptive mutant form of Gpa1, or the Msg5 dually specific phosphatase. The effects of Gpa1 and Msg5 on Fus3 are partially interdependent. In a genetic screen for adaptive defective mutants, a nonsense allele of the nucleocytoplasmic transport receptor, Kap104, was identified. Truncation of the Kap104 cargo binding domain blocked the effect of both Gpa1(E364K) and Msg5 on Fus3-GFP localization. Based on these results, we propose that Gpa1 and Msg5 work in concert to downregulate the mating signal and that they do so by inhibiting the pheromone-induced increase of Fus3 in the nucleus. Kap104 is required for the G(alpha)/phosphatase-mediated effect on Fus3 localization. PMID- 12556476 TI - Loss of HR6B ubiquitin-conjugating activity results in damaged synaptonemal complex structure and increased crossing-over frequency during the male meiotic prophase. AB - The ubiquitin-conjugating enzymes HR6A and HR6B are the two mammalian homologs of Saccharomyces cerevisiae RAD6. In yeast, RAD6 plays an important role in postreplication DNA repair and in sporulation. HR6B knockout mice are viable, but spermatogenesis is markedly affected during postmeiotic steps, leading to male infertility. In the present study, increased apoptosis of HR6B knockout primary spermatocytes was detected during the first wave of spermatogenesis, indicating that HR6B performs a primary role during the meiotic prophase. Detailed analysis of HR6B knockout pachytene nuclei showed major changes in the synaptonemal complexes. These complexes were found to be longer. In addition, we often found depletion of synaptonemal complex proteins from near telomeric regions in the HR6B knockout pachytene nuclei. Finally, we detected an increased number of foci containing the mismatch DNA repair protein MLH1 in these nuclei, reflecting a remarkable and consistent increase (20 to 25%) in crossing-over frequency. The present findings reveal a specific requirement for the ubiquitin-conjugating activity of HR6B in relation to dynamic aspects of the synaptonemal complex and meiotic recombination in spermatocytes. PMID- 12556477 TI - Small Maf compound mutants display central nervous system neuronal degeneration, aberrant transcription, and Bach protein mislocalization coincident with myoclonus and abnormal startle response. AB - The small Maf proteins form heterodimers with CNC and Bach family proteins to elicit transcriptional responses from Maf recognition elements (MAREs). We previously reported germ line-targeted deficiencies in mafG plus mafK compound mutant mice. The most prominent mutant phenotype was a progressive maf dosage dependent neuromuscular dysfunction. However, there has been no previous report regarding the effects of altered small-maf gene expression on neurological dysfunction. We show here that MafG and MafK are expressed in discrete central nervous system (CNS) neurons and that mafG::mafK compound mutants display neuronal degeneration coincident with surprisingly selective MARE-dependent transcriptional abnormalities. The CNS morphological changes are concurrent with the onset of a neurological disorder in the mutants, and the behavioral changes are accompanied by reduced glycine receptor subunit accumulation. Bach/small Maf heterodimers, which normally generate transcriptional repressors, were significantly underrepresented in nuclear extracts prepared from maf mutant brains, and Bach proteins fail to accumulate normally in nuclei. Thus compound mafG::mafK mutants develop age- and maf gene dosage-dependent cell-autonomous neuronal deficiencies that lead to profound neurological defects. PMID- 12556478 TI - Targeted disruption of the PDZK1 gene by homologous recombination. AB - Proteins containing PDZ domains are involved in a large number of biological functions, including protein scaffolding, organization of ion channels, and signal transduction. We recently identified a novel PDZ domain-containing protein, PDZK1, that is selectively expressed in normal tissues, where it is associated and colocalized with MAP17, a small 17-kDa membrane-associated protein; cMOAT, an organic anion transporter implicated in multidrug resistance; and the type IIa Na/Pi cotransporter. The protein cluster formed by PDZK1, MAP17, and cMOAT is upregulated in a significant number of human carcinomas originating in the colon, breast, lung, and kidney. In order to better define the function of PDZK1 in the protein cluster and its potential role in the organization of ion channels, we generated a PDZK1 knockout mouse. While PDZK1-deficient mice developed normally, did not display any gross phenotypic abnormalities, and were fecund, lack of PDZK1 resulted in modulation of expression of selective ion channels in the kidney, as well as increased serum cholesterol levels. However, no significant redistribution of proteins known to interact with PDZK1, such as MAP17, cMOAT, and the type IIa Na/Pi cotransporter, was observed. The absence of a more significant phenotype in PDZK1-deficient mice may be due to functional compensation by other PDZ domain-containing proteins, which could be instrumental in determining the location of interacting proteins such as ion channels and other membrane-associated proteins in defined areas of the plasma membrane. PMID- 12556479 TI - batman Interacts with polycomb and trithorax group genes and encodes a BTB/POZ protein that is included in a complex containing GAGA factor. AB - Polycomb and trithorax group genes maintain the appropriate repressed or activated state of homeotic gene expression throughout Drosophila melanogaster development. We have previously identified the batman gene as a Polycomb group candidate since its function is necessary for the repression of Sex combs reduced. However, our present genetic analysis indicates functions of batman in both activation and repression of homeotic genes. The 127-amino-acid Batman protein is almost reduced to a BTB/POZ domain, an evolutionary conserved protein protein interaction domain found in a large protein family. We show that this domain is involved in the interaction between Batman and the DNA binding GAGA factor encoded by the Trithorax-like gene. The GAGA factor and Batman codistribute on polytene chromosomes, coimmunoprecipitate from nuclear embryonic and larval extracts, and interact in the yeast two-hybrid assay. Batman, together with the GAGA factor, binds to MHS-70, a 70-bp fragment of the bithoraxoid Polycomb response element. This binding, like that of the GAGA factor, requires the presence of d(GA)n sequences. Together, our results suggest that batman belongs to a subset of the Polycomb/trithorax group of genes that includes Trithorax-like, whose products are involved in both activation and repression of homeotic genes. PMID- 12556480 TI - Potentiation of protein kinase C zeta activity by 15-deoxy-delta(12,14) prostaglandin J(2) induces an imbalance between mitogen-activated protein kinases and NF-kappa B that promotes apoptosis in macrophages. AB - Activation of the macrophage cell line RAW 264.7 with lipopolysaccharide (LPS) transiently activates protein kinase C zeta (PKC zeta) and Jun N-terminal kinase (JNK) through a phosphoinositide-3-kinase (PI3-kinase)-dependent pathway. Incubation of LPS-treated cells with the cyclopentenone 15-deoxy-Delta(12,14) prostaglandin J(2) (15dPGJ(2)) promoted a sustained activation of PKC zeta and JNK and inhibited I kappa B kinase (IKK) and NF-kappa B activity. Accordingly, 15dPGJ(2) induced an imbalance between JNK and IKK activities by increasing the former signaling pathway and inhibiting the latter signaling pathway. Under these conditions, apoptosis was significantly enhanced; this response was very dependent on PKC zeta and JNK activation. The effect of 15dPGJ(2) on PKC zeta activity observed in LPS-activated macrophages was not dependent on a direct action of this prostaglandin on the enzyme but was due to the activation of a step upstream of PI3-kinase. Moreover, LPS promoted the redistribution of activated PKC zeta from the cytosol to the nucleus, a process that was enhanced by treatment of the cells with 15dPGJ(2) that favored a persistent and broader distribution of PKC zeta in the nucleus. These results indicate that 15dPGJ(2) and other cyclopentenone prostaglandins, through the sustained activation of PKC zeta, might contribute significantly to the process of resolution of inflammation by promoting apoptosis of activated macrophages. PMID- 12556481 TI - SKIP negatively regulates insulin-induced GLUT4 translocation and membrane ruffle formation. AB - Skeletal muscle and kidney enriched inositol phosphatase (SKIP) is an inositol polyphosphate 5-phosphatase that hydrolyzes phosphatidylinositol 3,4,5 trisphosphate [PI(3,4,5)P3] to downregulate intracellular levels. In this study, we show that SKIP inhibits phosphoinositide 3-kinase signaling in insulin stimulated CHO cells. Ectopic expression of SKIP did not inhibit insulin-induced PI(3,4,5)P3 generation but did rapidly decrease insulin-induced intracellular PI(3,4,5)P3 levels compared with those in control cells. Further, insulin-induced phosphorylation of some downstream targets such as Akt and p70 S6 kinase was markedly inhibited by the ectopic expression of SKIP, whereas phosphorylation of mitogen-activated protein kinase was not. In contrast, downregulation of intracellular SKIP levels by antisense oligonucleotides dramatically enhanced Akt (protein kinase B) phosphorylation in response to insulin, suggesting that endogenous SKIP downregulates insulin signaling. SKIP also markedly inhibited GLUT4 translocation and membrane ruffle formation. We conclude that SKIP preferentially regulates glucose transport and actin cytoskeletal rearrangement among a variety of PI(3,4,5)P3 downstream events. PMID- 12556482 TI - Targeted disruption of the meprin beta gene in mice leads to underrepresentation of knockout mice and changes in renal gene expression profiles. AB - Meprins are multidomain zinc metalloproteases that are highly expressed in mammalian kidney and intestinal brush border membranes and in leukocytes and certain cancer cells. Mature meprins are oligomers of evolutionarily related, separately encoded alpha and/or beta subunits. Homooligomers of meprin alpha are secreted; oligomers containing meprin beta are plasma membrane associated. Meprin substrates include bioactive peptides and extracellular matrix proteins. Meprins have been implicated in cancer and intestinal inflammation. Additionally, meprin beta is a candidate gene for diabetic nephropathy. To elucidate in vivo functions of these metalloproteases, meprin beta null mice were generated by targeted disruption of the meprin beta gene on mouse chromosome 18q12. Analyses of meprin beta knockout mice indicated that (i) 50% fewer null mice are born than the Mendelian distribution predicts, (ii) null mice that survive develop normally and are viable and fertile, (iii) meprin beta knockout mice lack membrane-associated meprin alpha in kidney and intestine, and (iv) null mice have changes in renal gene expression profiles compared to wild-type mice as assessed by microarray analyses. Thus, disruption of the meprin beta allele in mice affects embryonic viability, birth weight, renal gene expression profiles, and the distribution of meprin alpha in kidney and intestine. PMID- 12556483 TI - TAB2 is essential for prevention of apoptosis in fetal liver but not for interleukin-1 signaling. AB - The proinflammatory cytokine interleukin-1 (IL-1) transmits a signal via several critical cytoplasmic proteins such as MyD88, IRAKs and TRAF6. Recently, serine/threonine kinase TAK1 and TAK1 binding protein 1 and 2 (TAB1/2) have been identified as molecules involved in IL-1-induced TRAF6-mediated activation of AP 1 and NF-kappa B via mitogen-activated protein (MAP) kinases and I kappa B kinases, respectively. However, their physiological functions remain to be clarified. To elucidate their roles in vivo, we generated TAB2-deficient mice. The TAB2 deficiency was embryonic lethal due to liver degeneration and apoptosis. This phenotype was similar to that of NF-kappa B p65-, IKK beta-, and NEMO/IKK gamma-deficient mice. However, the IL-1-induced activation of NF-kappa B and MAP kinases was not impaired in TAB2-deficient embryonic fibroblasts. These findings demonstrate that TAB2 is essential for embryonic development through prevention of liver apoptosis but not for the IL-1 receptor-mediated signaling pathway. PMID- 12556484 TI - Human Nudel and NudE as regulators of cytoplasmic dynein in poleward protein transport along the mitotic spindle. AB - Emerging evidence supports the idea that a signaling pathway containing orthologs of at least mammalian NudE and Nudel, Lis1, and cytoplasmic dynein is conserved for eukaryotic nuclear migration. In mammals, this pathway has profound impact on neuronal migration during development of the central nervous system. Lis1 and dynein are also involved in other cellular functions, such as mitosis. Here we show that Nudel also participates in a subset of dynein function in M phase. Nudel was specifically phosphorylated in M phase in its serine/threonine phosphorylation motifs, probably by Cdc2 and also Erk1 and -2. A fraction of Nudel bound to centrosomes strongly in interphase and localized to mitotic spindles in early M phase. By using mutants incapable of or simulating phosphorylation, we confirmed that phosphorylation of Nudel regulated the cell cycle-dependent distribution, possibly by increasing its dissociation rate at the microtubule-organizing center. Moreover, phosphorylated Nudel or the phosphorylation-mimicking mutant bound Lis1 more efficiently. We further demonstrated that a Nudel mutant incapable of binding to Lis1 impaired the poleward movement of dynein and hence the dynein-mediated transport of kinetochore proteins to spindle poles along microtubules, a process contributing to inactivation of the spindle checkpoint in mitosis. These results point to the importance of Nudel-Lis1 interaction for the dynein activity in M phase and to a possible role of Nudel phosphorylation as facilitating such interaction. In addition, comparative studies suggest that NudE is also functionally related to its paralog, Nudel. PMID- 12556485 TI - Impaired hepatocyte DNA synthetic response posthepatectomy in insulin-like growth factor binding protein 1-deficient mice with defects in C/EBP beta and mitogen activated protein kinase/extracellular signal-regulated kinase regulation. AB - After a two-thirds hepatectomy, normally quiescent liver cells are stimulated to reenter the cell cycle and proliferate to restore the original liver mass. One of the most rapidly and highly induced genes and proteins in regenerating liver is insulin-like growth factor binding protein 1 (IGFBP-1), a secreted protein that may modulate the activities of insulin-like growth factors (IGFs) or signal via IGF-independent mechanisms. To assess the functional role of IGFBP-1 in liver regeneration, mice with a targeted disruption of the IGFBP-1 gene were generated. Although IGFBP-1(-/-) mice demonstrated normal development, they had abnormal liver regeneration after partial hepatectomy, characterized by liver necrosis and reduced and delayed hepatocyte DNA synthesis. The abnormal regenerative response was associated with blunted activation of mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) and a reduced induction of C/EBP beta protein expression posthepatectomy. Like cell cycle abnormalities observed in hepatectomized C/EBP beta(-/-) mice, cyclin A and cyclin B1 expression was delayed and reduced in IGFBP-1(-/-) livers, whereas cyclin D1 expression was normal. Treatment of IGFBP-1(-/-) mice with a preoperative dose of IGFBP-1 induced MAPK/ERK activation and C/EBP beta expression, suggesting that IGFBP-1 may support liver regeneration at least in part via its effect on MAPK/ERK and C/EBP beta activities. These findings are the first demonstration of the involvement of IGFBP-1 in the regulation of in vivo mitogenic signaling pathways. PMID- 12556486 TI - Inactivation of the nuclear receptor coactivator RAP250 in mice results in placental vascular dysfunction. AB - Coactivators constitute a diverse group of proteins that are essential for optimal transcriptional activity of nuclear receptors. In the past few years many coactivators have been identified but it is still unclear whether these proteins interact indiscriminately with all nuclear receptors and whether there is some redundancy in their functions. We have previously cloned and characterized RAP250 (ASC-2/PRIP/TRBP/NRC), an LXXLL-containing coactivator for nuclear receptors. In order to study its biological role, Rap250 null mice were generated by gene targeting. Here we show that genetic disruption of Rap250 results in embryonic lethality at embryonic day (E) 13.5. Histological examination of placentas revealed a dramatically reduced spongiotrophoblast layer, a collapse of blood vessels in the region bordering the spongiotrophoblast, and labyrinthine layers in placentas from Rap250(-/-) embryos. These findings suggest that the lethality of Rap250(-/-) embryos is the result of obstructed placental blood circulation. Moreover, the transcriptional activity of PPAR gamma is reduced in fibroblasts derived from Rap250(-/-) embryos, suggesting that RAP250 is an essential coactivator for this nuclear receptor in the placenta. Our results demonstrate that RAP250 is necessary for placental development and thus essential for embryonic development. PMID- 12556487 TI - Haploinsufficiency of p18(INK4c) sensitizes mice to carcinogen-induced tumorigenesis. AB - The INK4 family of cyclin-dependent kinase (CDK) inhibitors negatively regulates cyclin D-dependent CDK4 and CDK6 and thereby retains the growth-suppressive function of Rb family proteins. Mutations in the CDK4 gene conferring INK4 resistance are associated with familial and sporadic melanoma in humans and result in a wide spectrum of tumors in mice. Whereas loss of function of other INK4 genes in mice leads to little or no tumor development, targeted deletion of p18(INK4c) causes spontaneous pituitary tumors and lymphoma late in life. Here we show that treatment of p18 null and heterozygous mice with a chemical carcinogen resulted in tumor development at an accelerated rate. The remaining wild-type allele of p18 was neither mutated nor silenced in tumors derived from heterozygotes. Hence, p18 is a haploinsufficient tumor suppressor in mice. PMID- 12556488 TI - Erythroid differentiation sensitizes K562 leukemia cells to TRAIL-induced apoptosis by downregulation of c-FLIP. AB - Regulation of the apoptotic threshold is of great importance in the homeostasis of both differentiating and fully developed organ systems. Triggering differentiation has been employed as a strategy to inhibit cell proliferation and accelerate apoptosis in malignant cells, in which the apoptotic threshold is often characteristically elevated. To better understand the mechanisms underlying differentiation-mediated regulation of apoptosis, we have studied death receptor responses during erythroid differentiation of K562 erythroleukemia cells, which normally are highly resistant to tumor necrosis factor (TNF) alpha-, FasL-, and TRAIL-induced apoptosis. However, upon hemin-mediated erythroid differentiation, K562 cells specifically lost their resistance to TNF-related apoptosis-inducing ligand (TRAIL), which efficiently killed the differentiating cells independently of mitochondrial apoptotic signaling. Concomitantly with the increased sensitivity, the expression of both c-FLIP splicing variants, c-FLIP(L) and c FLIP(S), was downregulated, resulting in an altered caspase 8 recruitment and cleavage in the death-inducing signaling complex (DISC). Stable overexpression of both c-FLIP(L) and c-FLIP(S) rescued the cells from TRAIL-mediated apoptosis with isoform-specific effects on DISC-recruited caspase 8. Our results show that c FLIP(L) and c-FLIP(S) potently control TRAIL responses, both by distinct regulatory features, and further imply that the differentiation state of malignant cells determines their sensitivity to death receptor signals. PMID- 12556489 TI - Growth arrest and DNA damage-inducible protein GADD34 targets protein phosphatase 1 alpha to the endoplasmic reticulum and promotes dephosphorylation of the alpha subunit of eukaryotic translation initiation factor 2. AB - The growth arrest and DNA damage-inducible protein, GADD34, associates with protein phosphatase 1 (PP1) and promotes in vitro dephosphorylation of the alpha subunit of eukaryotic translation initiation factor 2, (eIF-2 alpha). In this report, we show that the expression of human GADD34 in cultured cells reversed eIF-2 alpha phosphorylation induced by thapsigargin and tunicamycin, agents that promote protein unfolding in the endoplasmic reticulum (ER). GADD34 expression also reversed eIF-2 alpha phosphorylation induced by okadaic acid but not that induced by another phosphatase inhibitor, calyculin A (CA), which is a result consistent with PP1 being a component of the GADD34-assembled eIF-2 alpha phosphatase. Structure-function studies identified a bipartite C-terminal domain in GADD34 that encompassed a canonical PP1-binding motif, KVRF, and a novel RARA sequence, both of which were required for PP1 binding. N-terminal deletions of GADD34 established that while PP1 binding was necessary, it was not sufficient to promote eIF-2 alpha dephosphorylation in cells. Imaging of green fluorescent protein (GFP)-GADD34 proteins showed that the N-terminal 180 residues directed the localization of GADD34 at the ER and that GADD34 targeted the alpha isoform of PP1 to the ER. These data provide new insights into the mode of action of GADD34 in assembling an ER-associated eIF-2 alpha phosphatase that regulates protein translation in mammalian cells. PMID- 12556490 TI - Mouse germ cell-less as an essential component for nuclear integrity. AB - A mouse homologue of the Drosophila melanogaster germ cell-less (mgcl-1) gene is expressed ubiquitously, and its gene product is localized to the nuclear envelope based on its binding to LAP2 beta (lamina-associated polypeptide 2 beta). To elucidate the role of mgcl-1, we analyzed two mutant mouse lines that lacked mgcl 1 gene expression. Abnormal nuclear morphologies that were probably due to impaired nuclear envelope integrity were observed in the liver, exocrine pancreas, and testis. In particular, functional abnormalities were observed in testis in which the highest expression of mgcl-1 was detected. Fertility was significantly impaired in mgcl-1-null male mice, probably as a result of severe morphological abnormalities in the sperm. Electron microscopic observations showed insufficient chromatin condensation and abnormal acrosome structures in mgcl-1-null sperm. In addition, the expression patterns of transition proteins and protamines, both of which are essential for chromatin remodeling during spermatogenesis, were aberrant. Considering that the first abnormality during the process of spermatogenesis was abnormal nuclear envelope structure in spermatocytes, the mgcl-1 gene product appears to be essential for appropriate nuclear-lamina organization, which in turn is essential for normal sperm morphogenesis and chromatin remodeling. PMID- 12556492 TI - Formation of a tissue-specific histone acetylation pattern by the hematopoietic transcription factor GATA-1. AB - One function of lineage-restricted transcription factors may be to control the formation of tissue-specific chromatin domains. In erythroid cells, the beta globin gene cluster undergoes developmentally regulated hyperacetylation of histones at the active globin genes and the locus control region (LCR). However, it is unknown which transcription factor(s) governs the establishment of this erythroid-specific chromatin domain. We measured histone acetylation at the beta globin locus in the erythroid cell line G1E, which is deficient for the essential hematopoietic transcription factor GATA-1. Restoration of GATA-1 activity in G1E cells led to a substantial increase in acetylation of histones H3 and H4 at the beta-globin promoter and the LCR. Time course experiments showed that histone acetylation occurred rapidly after GATA-1 activation and coincided with globin gene expression, indicating that the effects of GATA-1 are direct. Moreover, increases in histone acetylation correlated with occupancy of GATA-1 and the acetyltransferase CBP at the locus in vivo. Together, these results suggest that GATA-1 and its cofactor CBP are essential for the formation of an erythroid specific acetylation pattern that is permissive for high levels of gene expression. PMID- 12556491 TI - Rho family GTPases are required for activation of Jak/STAT signaling by G protein coupled receptors. AB - As do cytokine receptors and receptor tyrosine kinases, G protein-coupled receptors (GPCRs) signal to Janus kinases (Jaks) and signal transducers and activators of transcription (STATs). However, the early biochemical events linking GPCRs to this signaling pathway have been unclear. Here we show that GPCR stimulated Rac activity and the subsequent generation of reactive oxygen species are necessary for activating tyrosine phosphorylation of Jaks and STAT-dependent transcription. The requirement for Rac activity can be overcome by addition of hydrogen peroxide. Expression of activated mutants of Rac1 is sufficient to activate Jak2 and STAT-dependent transcription, and the activation of Jak2 correlates with the ability of Rac1 to bind to NADPH oxidase subunit p67(phox). We further show that GPCR agonists stimulate tyrosine phosphorylation of STAT1 and STAT3 proteins in a Rac-dependent manner. The tyrosine phosphorylation of STAT3 is biphasic; the first peak of phosphorylation is weak and correlates with rapid activation of Jaks by GPCRs, whereas the second peak is stronger and requires the synthesis of an autocrine factor. Rho also plays an essential role in the induction of STAT transcriptional activity. Our results highlight a novel role for Rho GTPases in mediating the regulatory effects of GPCRs on STAT dependent gene expression. PMID- 12556493 TI - Snf1 kinases with different beta-subunit isoforms play distinct roles in regulating haploid invasive growth. AB - The Snf1 protein kinase of Saccharomyces cerevisiae has been shown to have a role in regulating haploid invasive growth in response to glucose depletion. Cells contain three forms of the Snf1 kinase, each with a different beta-subunit isoform, either Gal83, Sip1, or Sip2. We present evidence that different Snf1 kinases play distinct roles in two aspects of invasive growth, namely, adherence to the agar substrate and filamentation. The Snf1-Gal83 form of the kinase is required for adherence, whereas either Snf1-Gal83 or Snf1-Sip2 is sufficient for filamentation. Genetic evidence indicates that Snf1-Gal83 affects adherence by antagonizing Nrg1- and Nrg2-mediated repression of the FLO11 flocculin and adhesin gene. In contrast, the mechanism(s) by which Snf1-Gal83 and Snf1-Sip2 affect filamentation is independent of FLO11. Thus, the Snf1 kinase regulates invasive growth by at least two distinct mechanisms. PMID- 12556494 TI - Replication and expansion of trinucleotide repeats in yeast. AB - The mechanisms of trinucleotide repeat expansions, underlying more than a dozen hereditary neurological disorders, are yet to be understood. Here we looked at the replication of (CGG)(n) x (CCG)(n) and (CAG)(n) x (CTG)(n) repeats and their propensity to expand in Saccharomyces cerevisiae. Using electrophoretic analysis of replication intermediates, we found that (CGG)(n) x (CCG)(n) repeats significantly attenuate replication fork progression. Replication inhibition for this sequence becomes evident at as few as approximately 10 repeats and reaches a maximal level at 30 to 40 repeats. This is the first direct demonstration of replication attenuation by a triplet repeat in a eukaryotic system in vivo. For (CAG)(n) x (CTG)(n) repeats, on the contrary, there is only a marginal replication inhibition even at 80 repeats. The propensity of trinucleotide repeats to expand was evaluated in a parallel genetic study. In wild-type cells, expansions of (CGG)(25) x (CCG)(25) and (CAG)(25) x (CTG)(25) repeat tracts occurred with similar low rates. A mutation in the large subunit of the replicative replication factor C complex (rfc1-1) increased the expansion rate for the (CGG)(25) repeat approximately 50-fold but had a much smaller effect on the expansion of the (CTG)(25) repeat. These data show dramatic sequence-specific expansion effects due to a mutation in the lagging strand DNA synthesis machinery. Together, the results of this study suggest that expansions are likely to result when the replication fork attempts to escape from the stall site. PMID- 12556495 TI - Signal-induced transcriptional activation by Dif requires the dTRAP80 mediator module. AB - The Mediator complex is the major multiprotein transcriptional coactivator complex in Drosophila melanogaster. Mediator components interact with diverse sets of transcriptional activator proteins to elicit the sophisticated regulation of gene expression. The distinct phenotypes associated with certain mutations in some of the Mediator genes and the specific in vitro interactions of Mediator gene products with transcriptional activator proteins suggest the presence of activator-specific binding subunits within the Mediator complex. However, the physiological relevance of these selective in vitro interactions has not been addressed. Therefore, we analyzed dTRAP80, one of the putative activator-binding subunits of the Mediator, for specificity of binding to a number of natural transcriptional activators from Drosophila. Among the group of activator proteins that requires the Mediator complex for transcriptional activation, only a subset of these proteins interacted with dTRAP80 in vitro and only these dTRAP80 interacting activators were defective for activation under dTRAP80-deficient in vivo conditions. In particular, activation of Drosophila antimicrobial peptide drosomycin gene expression by the NF-kappa B-like transcription factor Dif during induction of the Toll signaling pathway was dependent on the dTRAP80 module. These results, and the indirect support from the dTRAP80 artificial recruitment assay, indicate that dTRAP80 serves as a genuine activator-binding target responsible for a distinct group of activators. PMID- 12556496 TI - Dual roles for Spt5 in pre-mRNA processing and transcription elongation revealed by identification of Spt5-associated proteins. AB - During transcription elongation, eukaryotic RNA polymerase II (Pol II) must contend with the barrier presented by nucleosomes. The conserved Spt4-Spt5 complex has been proposed to regulate elongation through nucleosomes by Pol II. To help define the mechanism of Spt5 function, we have characterized proteins that coimmunopurify with Spt5. Among these are the general elongation factors TFIIF and TFIIS as well as Spt6 and FACT, factors thought to regulate elongation through nucleosomes. Spt5 also coimmunopurified with the mRNA capping enzyme and cap methyltransferase, and spt4 and spt5 mutations displayed genetic interactions with mutations in capping enzyme genes. Additionally, we found that spt4 and spt5 mutations lead to accumulation of unspliced pre-mRNA. Spt5 also copurified with several previously unstudied proteins; we demonstrate that one of these is encoded by a new member of the SPT gene family. Finally, by immunoprecipitating these factors we found evidence that Spt5 participates in at least three Pol II complexes. These observations provide new evidence of roles for Spt4-Spt5 in pre mRNA processing and transcription elongation. PMID- 12556497 TI - Regulation of lymphoid enhancer factor 1/T-cell factor by mitogen-activated protein kinase-related Nemo-like kinase-dependent phosphorylation in Wnt/beta catenin signaling. AB - The Wnt/beta-catenin signaling pathway regulates many developmental processes by modulating gene expression. Wnt signaling induces the stabilization of cytosolic beta-catenin, which then associates with lymphoid enhancer factor and T-cell factor (LEF-1/TCF) to form a transcription complex that activates Wnt target genes. Previously, we have shown that a specific mitogen-activated protein (MAP) kinase pathway involving the MAP kinase kinase kinase TAK1 and MAP kinase-related Nemo-like kinase (NLK) suppresses Wnt signaling. In this study, we investigated the relationships among NLK, beta-catenin, and LEF-1/TCF. We found that NLK interacts directly with LEF-1/TCF and indirectly with beta-catenin via LEF-1/TCF to form a complex. NLK phosphorylates LEF-1/TCF on two serine/threonine residues located in its central region. Mutation of both residues to alanine enhanced LEF 1 transcriptional activity and rendered it resistant to inhibition by NLK. Phosphorylation of TCF-4 by NLK inhibited DNA binding by the beta-catenin-TCF-4 complex. However, this inhibition was abrogated when a mutant form of TCF-4 was used in which both threonines were replaced with valines. These results suggest that NLK phosphorylation on these sites contributes to the down-regulation of LEF 1/TCF transcriptional activity. PMID- 12556498 TI - Functional cross-antagonism between transcription factors FLI-1 and EKLF. AB - FLI-1 is an ETS family transcription factor which is overexpressed in Friend erythroleukemia and contributes to the blockage of differentiation of erythroleukemic cells. We show here that FLI-1 represses the transcriptional activity of the beta-globin gene promoter in MEL cells and interacts with two of its critical transactivators, GATA-1 and EKLF. Unexpectedly, FLI-1 enhances the stimulating activity of GATA-1 on a GATA-1-responsive promoter but represses that of EKLF on beta-globin and an EKLF-responsive artificial promoters. This repressive effect of FLI-1 requires the ETS DNA binding domain and its association with either the N- or C-terminal domain, which themselves interact with EKLF but not with GATA-1. Furthermore, the FLI-1 ETS domain alone behaves as an autonomous repression domain when linked to the Gal4 DNA binding domain. Taken together, these data indicate that FLI-1 represses EKLF-dependent transcription due to the repression activity of its ETS domain and its indirect recruitment to erythroid promoters by protein-protein interaction with EKLF. Reciprocally, we also show that EKLF itself represses the FLI-1-dependent megakaryocytic GPIX gene promoter, thus further suggesting that functional cross-antagonism between FLI-1 and EKLF might be involved in the control of the erythrocytic versus megakaryocytic differentiation of bipotential progenitors. PMID- 12556499 TI - Molecular dissection of mitotic recombination in the yeast Saccharomyces cerevisiae. AB - Recombination plays a central role in the repair of broken chromosomes in all eukaryotes. We carried out a systematic study of mitotic recombination. Using several assays, we established the chronological sequence of events necessary to repair a single double-strand break. Once a chromosome is broken, yeast cells become immediately committed to recombinational repair. Recombination is completed within an hour and exhibits two kinetic gaps. By using this kinetic framework we also characterized the role played by several proteins in the recombinational process. In the absence of Rad52, the broken chromosome ends, both 5' and 3', are rapidly degraded. This is not due to the inability to recombine, since the 3' single-stranded DNA ends are stable in a strain lacking donor sequences. Rad57 is required for two consecutive strand exchange reactions. Surprisingly, we found that the Srs2 helicase also plays an early positive role in the recombination process. PMID- 12556500 TI - Regulation of c-Rel nuclear localization by binding of Ca2+/calmodulin. AB - The NF-kappa B/Rel family of transcription factors participates in the control of a wide array of genes, including genes involved in embryonic development and regulation of immune, inflammation, and stress responses. In most cells, inhibitory I kappa B proteins sequester NF-kappa B/Rel in the cytoplasm. Cellular stimulation results in the degradation of I kappa B and modification of NF-kappa B/Rel proteins, allowing NF-kappa B/Rel to translocate to the nucleus and act on its target genes. Calmodulin (CaM) is a highly conserved, ubiquitously expressed Ca(2+) binding protein that serves as a key mediator of intracellular Ca(2+) signals. Here we report that two members of the NF-kappa B/Rel family, c-Rel and RelA, interact directly with Ca(2+)-loaded CaM. The interaction with CaM is greatly enhanced by cell stimulation, and this enhancement is blocked by addition of I kappa B. c-Rel and RelA interact with CaM through a similar sequence near the nuclear localization signal. Compared to the wild-type protein, CaM binding deficient mutants of c-Rel exhibit increases in both nuclear accumulation and transcriptional activity on the interleukin 2 and granulocyte macrophage colony stimulating factor promoters in the presence of a Ca(2+) signal. Conversely, for RelA neither nuclear accumulation nor transcriptional activity on these promoters is increased by mutation of the sequence interacting with CaM. Our results suggest that CaM binds c-Rel and RelA after their release from I kappa B and can inhibit nuclear import of c-Rel while letting RelA translocate to the nucleus and act on its target genes. CaM can therefore differentially regulate the activation of NF-kappa B/Rel proteins following stimulation. PMID- 12556501 TI - Two adjacent trimeric Fas ligands are required for Fas signaling and formation of a death-inducing signaling complex. AB - The membrane-bound form of Fas ligand (FasL) signals apoptosis in target cells through engagement of the death receptor Fas, whereas the proteolytically processed, soluble form of FasL does not induce cell death. However, soluble FasL can be rendered active upon cross-linking. Since the minimal extent of oligomerization of FasL that exerts cytotoxicity is unknown, we engineered hexameric proteins containing two trimers of FasL within the same molecule. This was achieved by fusing FasL to the Fc portion of immunoglobulin G1 or to the collagen domain of ACRP30/adiponectin. Trimeric FasL and hexameric FasL both bound to Fas, but only the hexameric forms were highly cytotoxic and competent to signal apoptosis via formation of a death-inducing signaling complex. Three sequential early events in Fas-mediated apoptosis could be dissected, namely, receptor binding, receptor activation, and recruitment of intracellular signaling molecules, each of which occurred independently of the subsequent one. These results demonstrate that the limited oligomerization of FasL, and most likely of some other tumor necrosis factor family ligands such as CD40L, is required for triggering of the signaling pathways. PMID- 12556502 TI - Direct kinase-to-kinase signaling mediated by the FHA phosphoprotein recognition domain of the Dun1 DNA damage checkpoint kinase. AB - The serine-threonine kinase Dun1 contains a forkhead-associated (FHA) domain and functions in the DNA damage checkpoint pathway of Saccharomyces cerevisiae. It belongs to the Chk2 family of checkpoint kinases, which includes S. cerevisiae Rad53 and Mek1, Schizosaccharomyces pombe Cds1, and human Chk2. Dun1 is required for DNA damage-induced transcription of certain target genes, transient G(2)/M arrest after DNA damage, and DNA damage-induced phosphorylation of the DNA repair protein Rad55. Here we report that the FHA phosphoprotein recognition domain of Dun1 is required for direct phosphorylation of Dun1 by Rad53 kinase in vitro and in vivo. trans phosphorylation by Rad53 does not require the Dun1 kinase activity and is likely to involve only a transient interaction between the two kinases. The checkpoint functions of Dun1 kinase in DNA damage-induced transcription, G(2)/M cell cycle arrest, and Rad55 phosphorylation are severely compromised in an FHA domain mutant of Dun1. As a consequence, the Dun1 FHA domain mutant displays enhanced sensitivity to genotoxic stress induced by UV, methyl methanesulfonate, and the replication inhibitor hydroxyurea. We show that the Dun1 FHA domain is critical for direct kinase-to-kinase signaling from Rad53 to Dun1 in the DNA damage checkpoint pathway. PMID- 12556503 TI - Yeast DNA polymerase zeta is an efficient extender of primer ends opposite from 7,8-dihydro-8-Oxoguanine and O6-methylguanine. AB - Genetic studies in Saccharomyces cerevisiae have indicated the requirement of DNA polymerase (Pol) zeta for mutagenesis induced by UV light and by other DNA damaging agents. However, on its own, Pol zeta is highly inefficient at replicating through DNA lesions; rather, it promotes their mutagenic bypass by extending from the nucleotide inserted opposite the lesion by another DNA polymerase. So far, such a role for Pol zeta has been established for cyclobutane pyrimidine dimers, (6-4) dipyrimidine photoproducts, and abasic sites. Here, we examine whether Pol zeta can replicate through the 7,8-dihydro-8-oxoguanine (8 oxoG) and O(6)-methylguanine (m6G) lesions. We chose these two lesions for this study because the replicative polymerase, Pol delta, can replicate through them, albeit weakly. We found that Pol zeta is very inefficient at inserting nucleotides opposite both these lesions, but it can efficiently extend from the nucleotides inserted opposite them by Pol delta. Also, the most efficient bypass of 8-oxoG and m6G lesions occurs when Pol delta is combined with Pol zeta, indicating a role for Polzeta in extending from the nucleotides inserted opposite these lesions by Pol delta. Thus, Pol zeta is a highly specialized polymerase that can proficiently extend from the primer ends opposite DNA lesions, irrespective of their degree of geometric distortion. Pol zeta, however, is unusually sensitive to geometric distortion of the templating residue, as it is highly inefficient at incorporating nucleotides even opposite the moderately distorting 8-oxoG and m6G lesions. PMID- 12556504 TI - Maintenance of open chromatin and selective genomic occupancy at the cell cycle regulated histone H4 promoter during differentiation of HL-60 promyelocytic leukemia cells. AB - During the shutdown of proliferation and onset of differentiation of HL-60 promyelocytic leukemia cells, expression of the cell cycle-dependent histone genes is downregulated at the level of transcription. To address the mechanism by which this regulation occurs, we examined the chromatin structure of the histone H4/n (FO108, H4FN) gene locus. Micrococcal nuclease, DNase I, and restriction enzymes show similar cleavage sites and levels of sensitivity at the H4/n locus in both proliferating and differentiated HL-60 cells. In contrast, differentiation-related activation of the cyclin-dependent kinase inhibitor p21(cip1/WAF1) gene is accompanied by increased nuclease hypersensitivity. Chromatin immunoprecipitation assays of the H4/n gene reveal that acetylated histones H3 and H4 are maintained at the same levels in proliferating and postproliferative cells. Thus, the chromatin of the H4/n locus remains in an open state even after transcription ceases. Using ligation-mediated PCR to visualize genomic DNase I footprints at single-nucleotide resolution, we find that protein occupancy at the site II cell cycle element is selectively diminished in differentiated cells while the site I element remains occupied. Decreased occupancy of site II is reflected by loss of the site II binding protein HiNF-P. We conclude that H4 gene transcription during differentiation is downregulated by modulating protein interaction at the site II cell cycle element and that retention of an open chromatin conformation may be associated with site I occupancy. PMID- 12556507 TI - Society of pediatric psychology task force report: recommendations for the training of pediatric psychologists. AB - OBJECTIVE: To provide an overview of the types of training experiences considered most important to the development of competency in pediatric psychology. METHODS: This is the work of a task force commissioned by the Society of Pediatric Psychology, Division 54 of the American Psychological Association. RESULTS: Twelve topic areas, adapted from Roberts et al. (1998), deemed important for obtaining knowledge and expertise in pediatric psychology, were identified. These topics include life span developmental psychology; life span developmental psychopathology; child, adolescent, and family assessment; intervention strategies; research methods and systems evaluation; professional, ethical, and legal issues pertaining to children, adolescents, and families; issues of diversity; the role of multiple disciplines in service-delivering systems; prevention, family support, and health promotion; social issues affecting children, adolescents, and families; consultant and liaison roles; and disease process and medical management. Each area is briefly described and recommendations for obtaining training in these areas are offered. CONCLUSIONS: The Society of Pediatric Psychology offers this document as a comprehensive review of the ideal types of training experiences most important to developing competencies in pediatric psychology. These recommendations can be used by graduate students and graduate programs in shaping a training plan for students interested in pediatric psychology training. PMID- 12556505 TI - Disruption of the pelota gene causes early embryonic lethality and defects in cell cycle progression. AB - Mutations in either the Drosophila melanogaster pelota or pelo gene or the Saccharomyces cerevisiae homologous gene, DOM34, cause defects of spermatogenesis and oogenesis in Drosophila, and delay of growth and failure of sporulation in yeast. These phenotypes suggest that pelota is required for normal progression of the mitotic and meiotic cell cycle. To determine the role of the pelota in mouse development and progression of cell cycle, we have established a targeted disruption of the mouse PELO: Heterozygous animals are variable and fertile. Genotyping of the progeny of heterozygous intercrosses shows the absence of Pelo( /-) pups and suggests an embryo-lethal phenotype. Histological analyses reveal that the homozygous Pelo deficient embryos fail to develop past day 7.5 of embryogenesis (E7.5). The failure of mitotic active inner cell mass of the Pelo( /-) blastocysts to expand in growth after 4 days in culture and the survival of mitotic inactive trophoplast indicate that the lethality of Pelo-null embryos is due to defects in cell proliferation. Analysis of the cellular DNA content reveals the significant increase of aneuploid cells in Pelo(-/-) embryos at E7.5. Therefore, the percent increase of aneuploid cells at E7.5 may be directly responsible for the arrested development and suggests that Pelo is required for the maintenance of genomic stability. PMID- 12556508 TI - Predoctoral training in pediatric psychology at the University of Kansas Clinical Child Psychology Program. PMID- 12556509 TI - Commentary: internship training. PMID- 12556510 TI - Commentary: recommendations for the training of pediatric psychologists: implications for postdoctoral training. PMID- 12556511 TI - Training experiences and theoretical orientations of pediatric psychologists. AB - OBJECTIVE: To reexamine members of the Society of Pediatric Psychology (SPP) to document trends in training experiences of individuals who identify themselves as pediatric psychologists. METHODS: Using the 1999 SPP membership list, we asked respondents to complete a survey describing their degree status, theoretical orientation of graduate program, type of internship/fellowship, current employment setting, and orientation to assessment and treatment of pediatric problems. RESULTS: The majority of the respondents matriculated from doctoral programs in the last two decades. Compared to a previous survey (Mullins, Harbeck Weber, Olson, & Hartman, 1996), this survey had a higher percentage of respondents who completed accredited predoctoral or postdoctoral training in pediatric psychology. Over half of the respondents were primarily employed in medical centers or academic institutions; fewer respondents reported working in private practice settings than in the previous survey 10 years ago. Respondents' current orientation was most often described as cognitive-behavioral. In this survey, the percentage of respondents endorsing an eclectic orientation was lower; the percentage endorsing cognitive-behavioral, behavioral, and systems orientations increased. CONCLUSIONS: We discuss implications of these findings as they relate to training and practice in pediatric psychology. PMID- 12556512 TI - Training graduate-level pediatric psychology researchers at Case Western Reserve University: meeting the challenges of the new millennium. AB - OBJECTIVE: To describe the challenges in training graduate-level pediatric psychology researchers for successful careers and to discuss solutions. METHODS: We reviewed experiences in training graduate students at Case Western Reserve University to identify key challenges in research training and potential strategies to meet them. RESULTS: We identified the following key challenges: stimulating graduate students' career interest in pediatric psychology research; teaching students about the pragmatic challenges of conducting research in pediatric settings, specialized research design, and data analytic issues; helping students to develop essential research skills; developing opportunities for student research-related collaborations; helping students develop professional identities as researchers; and developing and supporting their research careers beyond graduate school. CONCLUSIONS: Useful strategies for meeting these challenges include involving an interdisciplinary faculty in research training; developing specialized training methods that focus on critical research skills such as writing and data analysis; peer support; and involvement with multiple mentors who are successful researchers. Pediatric psychologists should also develop opportunities for the next generation of researchers by facilitating research job options and leadership opportunities. PMID- 12556513 TI - Pediatric psychology training and genetics: what will twenty-first-century pediatric psychologists need to know? AB - OBJECTIVE: To demonstrate the importance of genetic knowledge in coming decades and to outline necessary areas of genetic education. METHOD: This article reviews research involving genetic testing of children for cancer syndromes, development disabilities, psychiatric problems, and other conditions. RESULTS: The developmental, clinical, research, and consultation skills of well-trained pediatric psychologists will make them valuable collaborators with genetics professionals. Pediatric psychologists study the genetic etiology of psychiatric conditions and outcomes of genetic testing for physical disease. CONCLUSIONS: Pediatric psychologists will need training in the concepts and methods of the New Genetics. They should understand the implications of risk notification and genetic test disclosure and should be aware of related ethical concerns. PMID- 12556514 TI - Preparing psychologists to link systems of care in managing and preventing children's health problems. AB - OBJECTIVE: To describe the need for innovations in training to link health, educational, and family systems and to illustrate how this can be accomplished through child-oriented psychology training programs. METHODS: We describe multiple pathways for the preparation of child-oriented psychologists to link health, educational, and family systems, in keeping with the National Institute of Mental Health guidelines for preparing professionals in child and adolescent psychology. These pathways include training embedded in graduate programs specializing in clinical child, pediatric, school, community, and family psychology. This article highlights a training initiative for preparing child oriented psychologists based in a school psychology program. RESULTS: A partnership between Lehigh University and The Children's Hospital of Philadelphia has been developed to prepare school psychologists to coordinate community-based systems of care to promote positive educational and health outcomes for children. This program emphasizes both intervention and prevention and provides a set of integrated experiences in both health care and educational settings. CONCLUSIONS: We highlight components of this program relevant to the preparation of pediatric psychologists. We identify and discuss potential challenges in establishing training programs for the preparation of professionals to link health, school, and family systems. PMID- 12556516 TI - Induction of the ZRC1 metal tolerance gene in zinc-limited yeast confers resistance to zinc shock. AB - Zinc is an essential nutrient but toxic to cells with overaccumulation. For this reason, intracellular zinc levels are tightly controlled. In the yeast Saccharomyces cerevisiae, the Zrc1 and Cot1 proteins have been implicated in the storage and detoxification of excess zinc in the vacuole. Surprisingly, transcription of ZRC1 is induced in zinc-limited cells by the zinc-responsive transcription factor Zap1. We show here that this increase in ZRC1 expression is a novel mechanism of zinc homeostasis and stress tolerance. Zinc-limited cells also express high levels of the plasma membrane zinc uptake transporters. As a consequence, when zinc-limited cells are resupplied with small amounts of zinc, large quantities quickly accumulate in the cell, a condition we refer to as "zinc shock." We show here that ZRC1 and its induction in zinc-limited cells are required for resistance to this zinc shock. Experiments using the zinc-responsive fluorophore FuraZin-1 as an indicator of vacuolar zinc levels indicated that Zrc1 is required for the rapid transport of zinc into the vacuole during zinc shock. We also present evidence that cytosolic zinc rises to higher levels in cells unable to sequester this excess zinc. Thus, the increase in ZRC1 expression occurs prior to the zinc shock stress for which this induction is important. We propose that this "proactive" strategy of homeostatic regulation, such as we document here for ZRC1, may represent a common but largely unrecognized phenomenon. PMID- 12556517 TI - Molecular dissection of the contribution of negatively and positively charged residues in S2, S3, and S4 to the final membrane topology of the voltage sensor in the K+ channel, KAT1. AB - Voltage-dependent ion channels control changes in ion permeability in response to membrane potential changes. The voltage sensor in channel proteins consists of the highly positively charged segment, S4, and the negatively charged segments, S2 and S3. The process involved in the integration of the protein into the membrane remains to be elucidated. In this study, we used in vitro translation and translocation experiments to evaluate interactions between residues in the voltage sensor of a hyperpolarization-activated potassium channel, KAT1, and their effect on the final topology in the endoplasmic reticulum (ER) membrane. A D95V mutation in S2 showed less S3-S4 integration into the membrane, whereas a D105V mutation allowed S4 to be released into the ER lumen. These results indicate that Asp(95) assists in the membrane insertion of S3-S4 and that Asp(105) helps in preventing S4 from being releasing into the ER lumen. The charge reversal mutation, R171D, in S4 rescued the D105R mutation and prevented S4 release into the ER lumen. A series of constructs containing different C terminal truncations of S4 showed that Arg(174) was required for correct integration of S3 and S4 into the membrane. Interactions between Asp(105) and Arg(171) and between negative residues in S2 or S3 and Arg(174) may be formed transiently during membrane integration. These data clarify the role of charged residues in S2, S3, and S4 and identify posttranslational electrostatic interactions between charged residues that are required to achieve the correct voltage sensor topology in the ER membrane. PMID- 12556518 TI - Mitochondrial targeting of normal and mutant protoporphyrinogen oxidase. AB - We have investigated the signal sequence for mitochondrial transport of mutants (I12T, 78insC, IVS2-2a-->c, 338G-->C, R152C, 470A-->C, and L401F) and the wild type protoporphyrinogen oxidase (PPOX), which is the penultimate enzyme in the heme biosynthesis. We constructed the corresponding green fluorescent protein fusion proteins and studied their intracellular localization in COS-1 cells. We showed that 28 amino acids in the amino terminus of PPOX contain an independently functioning signal for mitochondrial targeting. The experiments with amino terminally truncated green fluorescent protein fusion proteins revealed that amino acids 25-477 of PPOX contained an additional mitochondrial targeting signal(s). We constructed a structural model for the interaction between the amino-terminal end of PPOX and the putative mitochondrial receptor protein Tom20. The model suggests that leucine and isoleucine residues Leu-8, Ile-12, and Leu-15 forming an alpha-helical hydrophobic motif, LXXXIXXL, were crucial for the recognition of the targeting signal. The validity of the model was tested using mutants L8Q, I12T, and L15Q disrupting the hydrophobic surface of the LXXXIXXL helix. The results from in vitro expression studies and molecular modeling were in accordance supporting the hypothesis that the recognition of the mitochondrial targeting signal is dependent on hydrophobic interactions between the targeting signal and the mitochondrial receptor. PMID- 12556519 TI - Casein kinase I epsilon enhances the binding of Dvl-1 to Frat-1 and is essential for Wnt-3a-induced accumulation of beta-catenin. AB - We demonstrate that Dvl-1, casein kinase I epsilon (CKI epsilon), and Frat-1 activate the Wnt signaling pathway cooperatively. The amino acid region 228-250 of Dvl-1 was necessary for its binding to Frat-1, and the interaction of Dvl-1 with Frat-1 was enhanced by CKI epsilon. Coexpression of Dvl-1 and Frat-1 caused accumulation of beta-catenin synergistically in L cells. Both proteins also activated the transcriptional activity of T-cell factor-4 (Tcf-4) synergistically in human embryonic kidney 293 cells, but coexpression of Dvl-1-(Delta 228-250), which lacks the amino acid region 228-250 from Dvl-1, and Frat-1 did not. Dvl-1, but not Dvl-1-(Delta 228-250), acted synergistically with CKI epsilon to activate Tcf-4. Depletion of CKI epsilon by double-stranded RNA interference in HeLa S3 cells led to the inhibition of Wnt-3a-induced phosphorylation of Dvl and the binding of Dvl-1 to Frat-1. Furthermore, depletion of CKI epsilon reduced the Wnt 3a-induced accumulation of beta-catenin, although it did not affect the basal level of beta-catenin. These results indicate that CKI epsilon-dependent phosphorylation of Dvl enhances the formation of a complex of Dvl-1 with Frat-1 and that this complex leads to the activation of the Wnt signaling pathway. PMID- 12556520 TI - UUUUUNU oligonucleotide stimulation of vaccinia virus early gene transcription termination, in trans. AB - Vaccinia virus early gene transcription termination requires the vaccinia termination factor (VTF), NPH I, a single stranded DNA-dependent ATPase, the virion form of RNA polymerase containing the Rap 94 subunit, and the signal UUUUUNU, which resides in the nascent mRNA, located 30 to 50 bases upstream from the poly(A) addition site. Evidence indicates that a required termination factor acts through binding to the UUUUUNU signal. To further investigate the function of UUUUUNU, the ability of UUUUUNU containing oligonucleotides to inhibit transcription termination was tested. A 22-mer RNA oligonucleotide containing a central U9 sequence exhibited sequence and concentration-dependent stimulation of premature transcription termination and transcript release, in trans. Activation of premature termination required VTF, NPH I, Rap 94, and ATP, demonstrating that the normal termination machinery was employed. Premature termination was not stimulated by RNA harboring a mutant UUUUUNU, demonstrating specificity. These data are consistent with a model in which a required termination factor is converted from an inactive to an active form by binding to a UUUUUNU containing oligonucleotide. The active termination factor then interacts with the ternary complex stimulating transcription termination through the normal mechanism, independent of the nascent mRNA sequence. PMID- 12556521 TI - Targeted disruption of the ATP2A1 gene encoding the sarco(endo)plasmic reticulum Ca2+ ATPase isoform 1 (SERCA1) impairs diaphragm function and is lethal in neonatal mice. AB - Mutations in the ATP2A1 gene, encoding isoform 1 of the sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA1), are one cause of Brody disease, characterized in humans by exercise-induced contraction of fast twitch (type II) skeletal muscle fibers. In an attempt to create a model for Brody disease, the mouse ATP2A1 gene was targeted to generate a SERCA1-null mutant mouse line. In contrast to humans, term SERCA1-null mice had progressive cyanosis and gasping respiration and succumbed from respiratory failure shortly after birth. The percentage of affected homozygote SERCA1(-/-) mice was consistent with predicted Mendelian inheritance. A survey of multiple organs from 10-, 15-, and 18-day embryos revealed no morphological abnormalities, but analysis of the lungs in term mice revealed diffuse congestion and epithelial hypercellularity and studies of the diaphragm muscle revealed prominent hypercontracted regions in scattered fibers and increased fiber size variability. The V(max) of Ca(2+) transport activity in mutant diaphragm and skeletal muscle was reduced by 80% compared with wild-type muscle, and the contractile response to electrical stimulation under physiological conditions was reduced dramatically in mutant diaphragm muscle. No compensatory responses were detected in analysis of mRNAs encoding other Ca(2+) handling proteins or of protein levels. Expression of ATP2A1 is largely restricted to type II fibers, which predominate in normal mouse diaphragm. The absence of SERCA1 in type II fibers, and the absence of compensatory increases in other Ca(2+) handling proteins, coupled with the marked increase in contractile function required of the diaphragm muscle to support postnatal respiration, can account for respiratory failure in term SERCA1-null mice. PMID- 12556522 TI - Defining the active site of Schizosaccharomyces pombe C-terminal domain phosphatase Fcp1. AB - Fcp1 is an essential protein serine phosphatase that dephosphorylates the C terminal domain (CTD) of RNA polymerase II. By testing the effects of serial N- and C-terminal deletions of the 723-amino acid Schizosaccharomyces pombe Fcp1, we defined a minimal phosphatase domain spanning amino acids 156-580. We employed site-directed mutagenesis (introducing 24 mutations at 14 conserved positions) to locate candidate catalytic residues. We found that alanine substitutions for Arg(223), Asp(258), Lys(280), Asp(297), and Asp(298) abrogated the phosphatase activity with either p-nitrophenyl phosphate or CTD-PO(4) as substrates. Structure-activity relationships were determined by introducing conservative substitutions at each essential position. Our results, together with previous mutational studies, highlight a constellation of seven amino acids (Asp(170), Asp(172), Arg(223), Asp(258), Lys(280), Asp(297), and Asp(298)) that are conserved in all Fcp1 orthologs and likely comprise the active site. Five of these residues (Asp(170), Asp(172), Lys(280), Asp(297), and Asp(298)) are conserved at the active site of T4 polynucleotide 3'-phosphatase, suggesting that Fcp1 and T4 phosphatase are structurally and mechanistically related members of the DXD phosphotransferase superfamily. PMID- 12556523 TI - Heterogeneity in control of mRNA stability by AU-rich elements. AB - AU-rich elements (AREs), located in the 3'-untranslated region of unstable cytokine and chemokine mRNAs, promote rapid decay of otherwise stable mRNAs and may mediate selective mRNA stabilization in response to stimulation with interleukin-1 (IL-1). AREs vary considerably, however, in both size and sequence context. To assess the heterogeneity involved in control of mRNA stability by ARE motifs, human mRNA sequences from IL-1alpha-stimulated HEK293 cells and T98G cells were screened for either instability or stability using both cDNA (950 ARE containing sequences) and Affymetrix oligonucleotide (U95Av2 GeneChip) array analysis. Although ARE-containing mRNAs exhibited a broad range of stability, IL 1alpha promoted stability in a subset of mRNAs that were unstable when transcriptionally induced by tumor necrosis factor alpha. Stabilization of granulocyte/macrophage-colony stimulating factor and IL-8 mRNAs by IL-1alpha was achieved only after 2 h of stimulation, required ongoing protein synthesis, and depended on the activation of p38 MAPK. In contrast, stabilization of Gro3 mRNA in response to IL-1alpha was achieved immediately and was insensitive to inhibitors of protein synthesis and p38 MAPK activation. In concert, these findings demonstrate that ARE sequences are functionally heterogeneous; only a subset of unstable mRNAs is sensitive to stabilization by IL-1alpha. Moreover, IL 1alpha promotes stabilization of unstable mRNAs through distinct mechanistic pathways that distinguish between specific mRNA sequences. PMID- 12556524 TI - The three insulin response sequences in the glucose-6-phosphatase catalytic subunit gene promoter are functionally distinct. AB - Glucose-6-phosphatase catalyzes the terminal step in the gluconeogenic and glycogenolytic pathways. In HepG2 cells, the maximum repression of basal glucose 6-phosphatase catalytic subunit (G6Pase) gene transcription by insulin requires two distinct promoter regions, designated A (located between -231 and -199) and B (located between -198 and -159), that together form an insulin response unit. Region A binds hepatocyte nuclear factor-1, which acts as an accessory factor to enhance the effect of insulin, mediated through region B, on G6Pase gene transcription. We have previously shown that region B binds the transcriptional activator FKHR (FOXO1a) in vitro. Chromatin immunoprecipitation assays demonstrate that FKHR also binds the G6Pase promoter in situ and that insulin inhibits this binding. Region B contains three insulin response sequences (IRSs), designated IRS 1, 2, and 3, that share the core sequence T(G/A)TTTT. However, detailed analyses reveal that these three G6Pase IRSs are functionally distinct. Thus, FKHR binds IRS 1 with high affinity and IRS 2 with low affinity but it does not bind IRS 3. Moreover, in the context of the G6Pase promoter, IRS 1 and 2, but not IRS 3, are required for the insulin response. Surprisingly, IRS 3, as well as IRS 1 and IRS 2, can each confer an inhibitory effect of insulin on the expression of a heterologous fusion gene, indicating that, in this context, a transcription factor other than FKHR, or its orthologs, can also mediate an insulin response through the T(G/A)TTTT motif. PMID- 12556525 TI - Collagen XVIII, a basement membrane heparan sulfate proteoglycan, interacts with L-selectin and monocyte chemoattractant protein-1. AB - Leukocyte infiltration during inflammation is mediated by the sequential actions of adhesion molecules and chemokines. By using a rat ureteral obstruction model, we showed previously that L-selectin plays an important role in leukocyte infiltration into the kidney. Here we report the purification, identification, and characterization of an L-selectin-binding heparan sulfate proteoglycan (HSPG) expressed in the rat kidney. Partial amino acid sequencing and Western blotting analyses showed that the L-selectin-binding HSPG is collagen XVIII, a basement membrane HSPG. The binding of L-selectin to isolated collagen XVIII was specifically inhibited by an anti-L-selectin monoclonal antibody, EDTA, treatment of the collagen XVIII with heparitinase or heparin but not by chemically desulfated heparin. A cell binding assay showed that the L-selectin-collagen XVIII interaction mediates cell adhesion. Interestingly, collagen XVIII also interacted with a chemokine, monocyte chemoattractant protein-1, and presented it to a monocytic cell line, THP-1, which enhanced the alpha(4)beta(1) integrin mediated binding of the THP-1 cells to vascular cell adhesion molecule-1. Thus, collagen XVIII may provide a link between selectin-mediated cell adhesion and chemokine-induced cellular activation and accelerate the progression of leukocyte infiltration in renal inflammation. PMID- 12556526 TI - Mastoparan selectively activates phospholipase D2 in cell membranes. AB - Both known isoforms of phospholipase (PL) D, PLD1 and PLD2, require phosphatidylinositol 4,5-bisphosphate for activity. However, PLD2 is fully active in the presence of this phospholipid, whereas PLD1 activation is dependent on additional factors such as ADP-ribosylation factor-1 (ARF-1) and protein kinase Calpha. We find that mastoparan, an activator of G(i) and mast cells, stimulates an intrinsic PLD activity, most likely PLD2, in fractions enriched in plasma membranes from rat basophilic leukemia 2H3 mast cells. Overexpression of PLD2, but not of PLD1, results in a large increase in the mastoparan-inducible PLD activity in membrane fractions, particularly those enriched in plasma membranes. As in previous studies, expressed PLD2 is localized primarily in the plasma membrane and PLD1 in granule membranes. Studies with pertussis toxin and other agents indicate that mastoparan stimulates PLD2 independently of G(i), ARF-1, protein kinase C, and calcium. Kinetic studies indicate that mastoparan interacts synergistically with phosphatidylinositol 4,5-bisphosphate and that oleate, itself a weak stimulant of PLD2 at low concentrations, is a competitive inhibitor of mastoparan stimulation of PLD2. Therefore, mastoparan may be useful for investigating the regulation of PLD2, particularly in view of the well studied molecular interactions of mastoparan with certain other strategic signaling proteins. PMID- 12556527 TI - The role of the presenilin-1 homologue gene sel-12 of Caenorhabditis elegans in apoptotic activities. AB - Many cases of autosomal dominant early onset familial Alzheimer's disease result from mutations in presenilin-1 (PS1). In this study, we examined the role of the PS1 homologue gene sel-12 of Caenorhabditis elegans under oxidative stress and clarified the sel-12-induced apoptosis. A genetic null allele mutant, sel 12(ar171), showed resistance to oxidative stress and prevented mitochondrial dysfunction-induced apoptosis. On the other hand, another allele mutant, sel 12(ar131), that carries a missense mutation showed a proapoptotic activity, which may be the result of a gain of function property. Also, sel-12(ar131)-induced apoptosis was ced-3- and ced-4-dependent. Dantrolene, which specifically inhibits Ca(2+) release from endoplasmic reticulum stores, prevents sel-12(ar131)-induced apoptosis. SEL-12, which is localized in the endoplasmic reticulum, may induce apoptosis through abnormal calcium release from the endoplasmic reticulum. Together, with the previous finding that human PS1 could substitute for SEL-12, these results suggest the similar involvement of PS1-inducing apoptosis under oxidative stress and mitochondrial dysfunction in the Alzheimer's Disease brain. PMID- 12556528 TI - Tyrphostin A23 inhibits internalization of the transferrin receptor by perturbing the interaction between tyrosine motifs and the medium chain subunit of the AP-2 adaptor complex. AB - Several intracellular membrane trafficking events are mediated by tyrosine containing motifs within the cytosolic domains of integral membrane proteins. Many such motifs conform to the consensus YXXPhi, where Phi represents a bulky hydrophobic residue. This motif interacts with the medium chain (mu) subunits of adaptor complexes that link the cytosolic domains of integral membrane proteins to the clathrin coat involved in vesicle formation. The YXXPhi motif is similar to motifs in which the tyrosine residue is phosphorylated by tyrosine kinases. Tyrphostins (structural analogs of tyrosine) are inhibitors of tyrosine kinases and function by binding to the active sites of the enzymes. We previously showed that, in vitro and in yeast two-hybrid interaction assays, some tyrphostins can inhibit the interaction between YXXPhi motifs and the mu2 subunit of the AP-2 adaptor complex (Crump, C., Williams, J. L., Stephens, D. J., and Banting, G. (1998) J. Biol. Chem. 273, 28073-28077). A23 is such a tyrphostin. We now show that molecular modeling of tyrphostin A23 into the tyrosine-binding pocket in mu2 provides a structural explanation for A23 being able to inhibit the interaction between YXXPhi motifs and mu2. Furthermore, we show that A23 inhibited the internalization of (125)I-transferrin in Heb7a cells without having any discernible effect on the morphology of compartments of the endocytic pathway. Control tyrphostins, active as inhibitors of tyrosine kinase activity, but incapable of inhibiting the YXXPhi motif/mu2 interaction, did not inhibit endocytosis. These data are consistent with A23 inhibition of the YXXPhi motif/mu2 interaction in intact cells and with the possibility that different tyrphostins may be used to inhibit specific membrane trafficking events in eukaryotic cells. PMID- 12556529 TI - Selective formation of ErbB-2/ErbB-3 heterodimers depends on the ErbB-3 affinity of epidermal growth factor-like ligands. AB - EGF-like growth factors activate their ErbB receptors by promoting receptor mediated homodimerization or, alternatively, by the formation of heterodimers with the orphan ErbB-2 through an as yet unknown mechanism. To investigate the selectivity in dimer formation by ligands, we have applied the phage display approach to obtain ligands with modified C-terminal residues that discriminate between ErbB-2 and ErbB-3 as dimerization partners. We used the epidermal growth factor/transforming growth factor alpha chimera T1E as the template molecule because it binds to ErbB-3 homodimers with low affinity and to ErbB-2/ErbB-3 heterodimers with high affinity. Many phage variants were selected with enhanced binding affinity for ErbB-3 homodimers, indicating that C-terminal residues contribute to the interaction with ErbB-3. These variants were also potent ligands for ErbB-2/ErbB-3 heterodimers despite negative selection for such heterodimers. In contrast, phage variants positively selected for binding to ErbB 2/ErbB-3 heterodimers but negatively selected for binding to ErbB-3 homodimers can be considered as "second best" ErbB-3 binders, which require ErbB-2 heterodimerization for stable complex formation. Our findings imply that epidermal growth factor-like ligands bind ErbB-3 through a multi-domain interaction involving at least both linear endings of the ligand. Apparently the ErbB-3 affinity of a ligand determines whether it can form only ErbB-2/ErbB-3 complexes or also ErbB-3 homodimers. Because no separate binding domain for ErbB 2 could be identified, our data support a model in which ErbB heterodimerization occurs through a receptor-mediated mechanism and not through bivalent ligands. PMID- 12556530 TI - Structure and kinetic properties of Paracoccus pantotrophus cytochrome cd1 nitrite reductase with the d1 heme active site ligand tyrosine 25 replaced by serine. AB - The 1.4-A crystal structure of the oxidized state of a Y25S variant of cytochrome cd(1) nitrite reductase from Paracoccus pantotrophus is described. It shows that loss of Tyr(25), a ligand via its hydroxy group to the iron of the d(1) heme in the oxidized (as prepared) wild-type enzyme, does not result in a switch at the c heme of the unusual bishistidinyl coordination to the histidine/methionine coordination seen in other conformations of the enzyme. The Ser(25) side chain is seen in two positions in the d(1) heme pocket with relative occupancies of approximately 7:3, but in neither case is the hydroxy group bound to the iron atom; instead, a sulfate ion from the crystallization solution is bound between the Ser(25) side chain and the heme iron. Unlike the wild-type enzyme, the Y25S mutant is active as a reductase toward nitrite, oxygen, and hydroxylamine without a reductive activation step. It is concluded that Tyr(25) is not essential for catalysis of reduction of any substrate, but that the requirement for activation by reduction of the wild-type enzyme is related to a requirement to drive the dissociation of this residue from the active site. The Y25S protein retains the d(1) heme less well than the wild-type protein, suggesting that the tyrosine residue has a role in stabilizing the binding of this cofactor. PMID- 12556531 TI - EhPgp5 mRNA stability is a regulatory event in the Entamoeba histolytica multidrug resistance phenotype. AB - The multidrug resistance (MDR) phenotype in Entamoeba histolytica is characterized by the overexpression of the EhPgp5 gene in trophozoites grown in high drug concentrations. Here we evaluated the role of EhPgp5 mRNA stability on MDR using actinomycin D. EhPgp5 mRNA from trophozoites growing without emetine had a half-life of 2.1 h, which augmented to 3.1 h in cells cultured with 90 microM and to 7.8 h with 225 microM emetine. Polyadenylation sites were detected at 118-, 156-, and 189-nucleotide (nt) positions of the EhPgp5 mRNA 3' untranslated region. Interestingly, trophozoites grown with 225 microM emetine exhibited an extra polyadenylation site at 19 nt. The 3'-untranslated region sequence is AU-rich and has putative consensus sequences for RNA-binding proteins. We detected a RNA-protein complex in a region that contains a polypyrimidine tract (142-159 nt) and a cytoplasmic polyadenylation element (146 154 nt). A longer poly(A) tail in the EhPgp5 mRNA was seen in trophozoites grown with 225 microM emetine. Emetine stress may affect factors involved in mRNA turnover, including polyadenylation/deadenylation proteins, which could induce changes in the EhPgp5 mRNA half-life and poly(A) tail length. Novel evidence on mechanisms participating in E. histolytica MDR phenotype is provided. PMID- 12556532 TI - Identification of the NF-E2-related factor-2-dependent genes conferring protection against oxidative stress in primary cortical astrocytes using oligonucleotide microarray analysis. AB - The antioxidant responsive element (ARE) mediates transcriptional regulation of phase II detoxification enzymes and antioxidant proteins such as NAD(P)H:quinone oxidoreductase (NQO1), glutathione S-transferases, and glutamate-cysteine ligase. In this study, we demonstrate that NF-E2-related factor-2 (Nrf2) plays a major role in transcriptional activation of ARE-driven genes and identify Nrf2 dependent genes by oligonucleotide microarray analysis using primary cortical astrocytes from Nrf2(+/+) and Nrf2(-/-) mice. Nrf2(-/-) astrocytes had decreased basal NQO1 activity and no induction by tert-butylhydroquinone compared with Nrf2(+/+) astrocytes. Similarly, both basal and induced levels of human NQO1-ARE luciferase expression in Nrf2(-/-) astrocytes were significantly lower than in Nrf2(+/+) astrocytes. Furthermore, human NQO1-ARE-luciferase expression in Nrf2( /-) astrocytes was restored by overexpression of Nrf2, whereas ARE activation in Nrf2(+/+) astrocytes was completely blocked by dominant-negative Nrf2. In addition, we observed that Nrf2-dependent genes protected primary astrocytes from H(2)O(2)- or platelet-activating factor-induced apoptosis. In support of these observations, we identified Nrf2-dependent genes encoding detoxification enzymes, glutathione-related proteins, antioxidant proteins, NADPH-producing enzymes, and anti-inflammatory genes using oligonucleotide microarrays. Proteins within these functional categories are vital to the maintenance and responsiveness of a cell defense system, suggesting that an orchestrated change in gene expression via Nrf2 and the ARE gives a synergistic protective effect against oxidative stress. PMID- 12556533 TI - Regulation of the interleukin-1-induced signaling pathways by a novel member of the protein phosphatase 2C family (PP2Cepsilon). AB - Although TAK1 signaling plays essential roles in eliciting cellular responses to interleukin-1 (IL-1), a proinflammatory cytokine, how the IL-1-TAK1 signaling pathway is positively and negatively regulated remains poorly understood. In this study, we investigated the possible role of a novel protein phosphatase 2C (PP2C) family member, PP2Cepsilon, in the regulation of the IL-1-TAK1 signaling pathway. PP2Cepsilon was composed of 303 amino acids, and the overall similarity of amino acid sequence between PP2Cepsilon and PP2Calpha was found to be 26%. Ectopic expression of PP2Cepsilon inhibited the IL-1- and TAK1-induced activation of mitogen-activated protein kinase kinase 4 (MKK4)-c-Jun N-terminal kinase or MKK3 p38 signaling pathway. PP2Cepsilon dephosphorylated TAK1 in vitro. Co immunoprecipitation experiments indicated that PP2Cepsilon associates stably with TAK1 and attenuates the binding of TAK1 to MKK4 or MKK6. Ectopic expression of a phosphatase-negative mutant of PP2Cepsilon, PP2Cepsilon(D/A), which acted as a dominant negative form, enhanced both the association between TAK1 and MKK4 or MKK6 and the TAK1-induced activation of an AP-1 reporter gene. The association between PP2Cepsilon and TAK1 was transiently suppressed by IL-1 treatment of the cells. Taken together, these results suggest that, in the absence of IL-1-induced signal, PP2Cepsilon contributes to keeping the TAK1 signaling pathway in an inactive state by associating with and dephosphorylating TAK1. PMID- 12556534 TI - Characterization of a heart-specific fatty acid transport protein. AB - Fatty acids are a major source of energy for cardiac myocytes. Changes in fatty acid metabolism have been implicated as causal in diabetes and cardiac disease. The mechanism by which long chain fatty acids (LCFAs) enter cardiac myocytes is not well understood but appears to occur predominantly by protein-mediated transport. Here we report the cloning, expression pattern, and subcellular localization of a novel member of the fatty acid transport protein (FATP) family termed FATP6. FATP6 is principally expressed in the heart where it is the predominant FATP family member. Similar to other FATPs, transient and stable transfection of FATP6 into 293 cells enhanced uptake of LCFAs. FATP6 mRNA was localized to cardiac myocytes by in situ hybridization. Immunofluorescence microscopy of FATP6 in monkey and murine hearts revealed that the protein is exclusively located on the sarcolemma. FATP6 was restricted in its distribution to areas of the plasma membrane juxtaposed with small blood vessels. In these membrane domains FATP6 also colocalizes with another molecule involved in LCFA uptake, CD36. These findings suggest that FATP6 is involved in heart LCFA uptake, in which it may play a role in the pathogenesis of lipid-related cardiac disorders. PMID- 12556535 TI - Type 1 insulin-like growth factor receptor (IGF-IR) signaling inhibits apoptosis signal-regulating kinase 1 (ASK1). AB - The type 1 insulin-like growth factor receptor (IGF-IR) is a receptor-tyrosine kinase that plays a critical role in signaling cell survival and proliferation. IGF-IR binding to its ligand, insulin-like growth factor (IGF-I) activates phosphoinositide 3-kinase (PI3K), promotes cell proliferation by activating the mitogen-activated protein kinase (MAPK) cascade, and blocks apoptosis by inducing the phosphorylation and inhibition of proapoptotic proteins such as BAD. Apoptosis signal-regulating kinase 1 (ASK1) is a MAP kinase kinase kinase (MAPKKK) that is required for c-Jun N-terminal kinase (JNK) and p38 activation in response to Fas and tumor necrosis factor (TNF) receptor stimulation, and for oxidative stress- and TNFalpha-induced apoptosis. The results presented here indicate that ASK1 forms a complex with the IGF-IR and becomes phosphorylated on tyrosine residue(s) in a manner dependent on IGF-IR activity. IGF-IR signaling inhibited ASK1 irrespective of TNFalpha-induced ASK1 activation and resulted in decreased ASK1-dependent JNK1 stimulation. Signaling through IGF-IR rescued cells from ASK1-induced apoptotic cell death in a manner independent of PI3K activity. These results indicate that IGF-IR signaling suppresses the ASK-1-mediated stimulation of JNK/p38 and the induction of programmed cell death. The simultaneous activation of MAP kinases and the inhibition of the stress-activated arm of the cascade by IGF-IR may constitute a potent proliferative signaling system and is possibly a mechanism by which IGF-I can stimulate growth and inhibit cell death in a wide variety of cell types and biological settings. PMID- 12556536 TI - Leptin activates the anandamide hydrolase promoter in human T lymphocytes through STAT3. AB - Physiological concentrations of leptin stimulate the activity of the endocannabinoid-degrading enzyme anandamide hydrolase (fatty acid amide hydrolase, FAAH) in human T lymphocytes up to approximately 300% over the untreated controls. Stimulation of FAAH occurred through up-regulation of gene expression at transcriptional and translational levels and involved binding of leptin to its receptor with an apparent dissociation constant (K(d)) of 1.95 +/- 0.14 nm and maximum binding (B(max)) of 392 +/- 8 fmol x mg protein(-1). Leptin binding to the receptor triggered activation of STAT3 but not STAT1 or STAT5 or the mitogen-activated protein kinases p38, p42, and p44. Peripheral lymphocytes of leptin knock-out (ob/ob) mice showed decreased FAAH activity and expression (approximately 25% of the wild-type littermates), which were reversed to control levels by exogenous leptin. Analysis of the FAAH promoter showed a cAMP-response element-like site, which is a transcriptional target of STAT3. Consistently, mutation of this site prevented FAAH activation by leptin in transient expression assays. Electrophoretic mobility shift and supershift assays further corroborated the promoter activity data. Taken together, these results suggest that leptin, by up-regulating the FAAH promoter through STAT3, enhances FAAH expression, thus tuning the immunomodulatory effects of anandamide. These findings might also have critical implications for human fertility. PMID- 12556537 TI - Lymphotoxin beta receptor induces sequential activation of distinct NF-kappa B factors via separate signaling pathways. AB - Lymphotoxin beta receptor (LTbetaR)-induced activation of NF-kappaB in mouse embryo fibroblasts was mediated by the classical pathway and by an alternative or second pathway. The classical pathway involved the IkappaB kinase (IKK)beta- and IKKgamma-dependent degradation of IkappaBalpha and resulted in the rapid but transient activation of primarily RelA-containing NF-kappaB dimers. The alternative or second pathway proceeded via NF-kappaB-inducing kinase (NIK)-, IKKalpha-, and protein synthesis-dependent processing of the inhibitory NF kappaB2 p100 precursor protein to the p52 form and resulted in a delayed but sustained activation of primarily RelB-containing NF-kappaB dimers. This second pathway was independent of the classical IKK complex, which is governed by its central IKKgamma regulatory subunit. The sequential engagement of two distinct pathways, coupled with the negative feedback inhibition of RelA complexes by NF kappaB-induced resynthesis of IkappaBalpha, resulted in a pronounced temporal change in the nature of the NF-kappaB activity during the course of stimulation. Initially dominant RelA complexes were replaced with time by RelB complexes. Therefore, the alternative activation path mediated by processing of p100 was necessary for sustained NF-kappaB activity in mouse embryo fibroblasts in response to LTbetaR stimulation. Based on the phenotype of mice deficient in various components of the LTbetaR-induced activation of p100 processing, we conclude that this pathway is critically involved in the function of stromal cells during the generation of secondary lymphoid organ microarchitectures. PMID- 12556538 TI - Endothelial barrier strengthening by activation of focal adhesion kinase. AB - Endothelial cell barrier (EC) properties regulate blood tissue fluid flux. To determine the role of endothelial-matrix interactions in barrier regulation, we induced cell shrinkage by exposing confluent endothelial monolayers to hyperosmolarity. The dominant effect of a 15-min hyperosmolar exposure was an increase in the trans-endothelial electrical resistance, indicating the induction of barrier strengthening. Hyperosmolar exposure also increased activity of focal adhesion kinase and E-cadherin accumulation at the cell periphery. Concomitantly, the density of actin filaments increased markedly. In EC monolayers stably expressing constitutively active or dominant negative isoforms of Rac1, the actin response to hyperosmolar exposure was enhanced or blocked, respectively, although the response in trans-endothelial resistance was unaffected, indicating that the endothelial barrier enhancement occurred independently of actin. However, in monolayers expressing a kinase-deficient mutant of focal adhesion kinase, the hyperosmolarity-induced increases in activity of focal adhesion and peripheral E cadherin enhancement were blocked and the induced increase of electrical resistance was markedly blunted. These findings indicate that in EC exposed to hyperosmolar challenge, the involvement of focal adhesion kinase was critical in establishing barrier strengthening. PMID- 12556539 TI - Calcitonin receptor-stimulating peptide, a new member of the calcitonin gene related peptide family. Its isolation from porcine brain, structure, tissue distribution, and biological activity. AB - We isolated a novel biologically active peptide, designated calcitonin receptor stimulating peptide (CRSP), from the acid extract of the porcine brain by monitoring cAMP production in the porcine kidney cell line LLC-PK(1). Determination of the amino acid sequence and cDNA analysis encoding a CRSP precursor showed that this peptide has approximately 60% identity in the amino acid sequence with human calcitonin gene-related peptide type-alpha (alphaCGRP), type-beta (betaCGRP), and porcine CGRP. Northern blot analysis and radioimmunoassay demonstrated that CRSP is expressed mainly in the thyroid gland and the central nervous system, in which the calcitonin receptor was abundantly expressed. Synthetic CRSP elicited a potent stimulatory effect on the cAMP production in LLC-PK(1) cells. Although it shows significant sequence similarity with CGRPs, this peptide did not elicit cAMP elevation in cells that endogenously expressed a CGRP receptor or an adrenomedullin receptor or were transfected with either of these recombinant receptors. Administration of CRSP into anesthetized rats did not alter the blood pressure but induced a transient decrease in the plasma calcium concentration. In fact, this peptide potently increased the intracellular cAMP concentration in COS-7 cells that expressed the recombinant calcitonin receptor. These unique properties indicate that CRSP is not a porcine counterpart of betaCGRP and probably elicits its biological effects via the calcitonin receptor. PMID- 12556540 TI - Off-pump coronary-artery bypass surgery. PMID- 12556541 TI - Multifactorial intervention and cardiovascular disease in patients with type 2 diabetes. AB - BACKGROUND: Cardiovascular morbidity is a major burden in patients with type 2 diabetes. In the Steno-2 Study, we compared the effect of a targeted, intensified, multifactorial intervention with that of conventional treatment on modifiable risk factors for cardiovascular disease in patients with type 2 diabetes and microalbuminuria. METHODS: The primary end point of this open, parallel trial was a composite of death from cardiovascular causes, nonfatal myocardial infarction, nonfatal stroke, revascularization, and amputation. Eighty patients were randomly assigned to receive conventional treatment in accordance with national guidelines and 80 to receive intensive treatment, with a stepwise implementation of behavior modification and pharmacologic therapy that targeted hyperglycemia, hypertension, dyslipidemia, and microalbuminuria, along with secondary prevention of cardiovascular disease with aspirin. RESULTS: The mean age of the patients was 55.1 years, and the mean follow-up was 7.8 years. The decline in glycosylated hemoglobin values, systolic and diastolic blood pressure, serum cholesterol and triglyceride levels measured after an overnight fast, and urinary albumin excretion rate were all significantly greater in the intensive therapy group than in the conventional-therapy group. Patients receiving intensive therapy also had a significantly lower risk of cardiovascular disease (hazard ratio, 0.47; 95 percent confidence interval, 0.24 to 0.73), nephropathy (hazard ratio, 0.39; 95 percent confidence interval, 0.17 to 0.87), retinopathy (hazard ratio, 0.42; 95 percent confidence interval, 0.21 to 0.86), and autonomic neuropathy (hazard ratio, 0.37; 95 percent confidence interval, 0.18 to 0.79). CONCLUSIONS: A target-driven, long-term, intensified intervention aimed at multiple risk factors in patients with type 2 diabetes and microalbuminuria reduces the risk of cardiovascular and microvascular events by about 50 percent. PMID- 12556542 TI - A comparison of on-pump and off-pump coronary bypass surgery in low-risk patients. AB - BACKGROUND: The performance of coronary bypass surgery without cardiopulmonary bypass ("off pump") may reduce perioperative morbidity and costs, but it is uncertain whether the outcome is similar to that involving the use of cardiopulmonary bypass ("on pump"). METHODS: In a multicenter, randomized trial, we randomly assigned 139 patients with predominantly single- or double-vessel coronary disease to on-pump surgery and 142 to off-pump surgery. Cardiac outcome and cost effectiveness were determined one year after surgery. The uncertainty surrounding the cost-effectiveness ratio (cost differences per quality-adjusted year of life gained) was addressed by bootstrapping. RESULTS: At one year, the rate of freedom from death, stroke, myocardial infarction, and coronary reintervention was 90.6 percent after on-pump surgery and 88.0 percent after off pump surgery (absolute difference, 2.6 percent; 95 percent confidence interval, - 4.6 to 9.8). Graft patency in a randomized subgroup of patients was 93 percent after on-pump surgery and 91 percent after off-pump surgery (absolute difference, 2.0 percent; 95 percent confidence interval, - 6.5 to 10.4). On-pump surgery was associated with $1,839 in additional direct costs per patient ($14,908 vs. $13,069--a difference of 14.1 percent) and an increase in quality-adjusted years of life of 0.83 as compared with 0.82 (difference, 0.01 year; 95 percent confidence interval, - 0.03 to 0.04). Off-pump surgery was more cost effective than on-pump surgery in 95 percent of bootstrap estimates. CONCLUSIONS: In low risk patients, there was no difference in cardiac outcome at one year between those who underwent on-pump bypass surgery and those who underwent off-pump surgery. Off-pump surgery was more cost effective. PMID- 12556544 TI - Images in clinical medicine. Progression of the lesion at the site of inoculation after smallpox vaccination. PMID- 12556543 TI - Peritoneal dialysis and epithelial-to-mesenchymal transition of mesothelial cells. AB - BACKGROUND: During continuous ambulatory peritoneal dialysis, the peritoneum is exposed to bioincompatible dialysis fluids that cause denudation of mesothelial cells and, ultimately, tissue fibrosis and failure of ultrafiltration. However, the mechanism of this process has yet to be elucidated. METHODS: Mesothelial cells isolated from effluents in dialysis fluid from patients undergoing continuous ambulatory peritoneal dialysis were phenotypically characterized by flow cytometry, confocal immunofluorescence, Western blotting, and reverse transcriptase polymerase chain reaction. These cells were compared with mesothelial cells from omentum and treated with various stimuli in vitro to mimic the transdifferentiation observed during continuous ambulatory peritoneal dialysis. Results were confirmed in vivo by immunohistochemical analysis performed on peritoneal-biopsy specimens. RESULTS: Soon after dialysis is initiated, peritoneal mesothelial cells undergo a transition from an epithelial phenotype to a mesenchymal phenotype with a progressive loss of epithelial morphology and a decrease in the expression of cytokeratins and E-cadherin through an induction of the transcriptional repressor snail. Mesothelial cells also acquire a migratory phenotype with the up-regulation of expression of alpha2 integrin. In vitro analyses point to wound repair and profibrotic and inflammatory cytokines as factors that initiate mesothelial transdifferentiation. Immunohistochemical studies of peritoneal-biopsy specimens from patients undergoing continuous ambulatory peritoneal dialysis demonstrate the expression of the mesothelial markers intercellular adhesion molecule 1 and cytokeratins in fibroblast-like cells entrapped in the stroma, suggesting that these cells stemmed from local conversion of mesothelial cells. CONCLUSIONS: Our results suggest that mesothelial cells have an active role in the structural and functional alteration of the peritoneum during peritoneal dialysis. The findings suggest potential targets for the design of new dialysis solutions and markers for the monitoring of patients. PMID- 12556545 TI - Images in clinical medicine. Cowpox infection. PMID- 12556546 TI - Clinical practice. Premenstrual dysphoric disorder. PMID- 12556547 TI - Case records of the Massachusetts General Hospital. Weekly Clinicopathological exercises. Case 4-2003. A 42-year-old woman with cough, fever, and abnormalities on thoracoabdominal computed tomography. PMID- 12556548 TI - Reducing cardiovascular risk in type 2 diabetes. PMID- 12556549 TI - Preventing the return of smallpox. PMID- 12556550 TI - Polymorphisms of adrenergic receptors and the risk of heart failure. PMID- 12556551 TI - Antiretroviral therapy during pregnancy and the risk of an adverse outcome. PMID- 12556552 TI - Sleep apnea. PMID- 12556553 TI - Adolescent depression. PMID- 12556554 TI - Case 33-2002: a 28-year-old woman with ocular inflammation, fever, and headache. PMID- 12556555 TI - Clinical-trial agreements between medical schools and industry. PMID- 12556556 TI - Heregulin is sufficient for the promotion of tumorigenicity and metastasis of breast cancer cells in vivo. AB - Resistance of breast carcinomas to hormonal therapy is a clinical obstacle for the treatment of breast cancer. The molecular mechanisms and the factors involved in the progression of tumors from an estrogen (E2)-dependent to an E2-independent phenotype are not entirely understood. Heregulin (HRG) is a pleiotropic growth factor that binds to the erbB family of receptors, which are correlated with breast cancer progression and an aggressive phenotype in the breast carcinomas overexpressing the receptors. Previous studies in transgenic mice have shown that HRG is sufficient to induce mammary gland transformation and proliferation in the presence of hormonal stimulation. However, these studies did not address the important issue of the E2 independence that is part of the progression of breast cancer. In this study, we investigated the role of HRG in E2 independence. We were able to determine that HRG up-regulation was sufficient for the development of mammary tumors in the absence of E2 stimulation, a situation that mimics the progression of the human disease. We demonstrated that in ovariectomized nude mice, HRG induced E2 independence and antiestrogen resistance and promoted metastasis and preneoplastic transformation of the adjacent mouse mammary tissue. We show that one of the mechanisms by which HRG achieves the aggressive phenotype may be mediated via an increase in activated mitogen-activated protein kinase, an increase in a matrix-degrading enzyme, MMP-9, and the overexpression of vascular endothelial growth factors. The up-regulation of these genes occurred in the absence of any additional stimulation, in an autocrine manner. Our data provide new insights into the mechanisms of breast cancer progression in vivo, and reinforce the important role that HRG plays in this process. PMID- 12556557 TI - Direct evidence that Bcr-Abl tyrosine kinase activity disrupts normal synergistic interactions between Kit ligand and cytokines in primary primitive progenitor cells. AB - We previously reported that chronic myelogenous leukemia (CML) primitive granulocyte-monocyte (GM) progenitors have a greatly reduced requirement for kit ligand (KL) to achieve optimal growth with granulocyte colony-stimulating factor (G-CSF) + granulocyte-monocyte colony-stimulating factor (GM-CSF). Conversely, others have demonstrated that unlike normal, CML CD34+ progenitors can proliferate in response to KL as a sole stimulus. To address these seemingly paradoxical findings, we examined the growth responses of CML CD34+ GM progenitors to various cytokines with and without a potent inhibitor of Bcr-Abl tyrosine kinase activity, PD173955. The heightened growth responses of CML GM progenitors to KL alone and to G-CSF + GM-CSF were abrogated by 10 nM PD173955 while having no effect on normal GM progenitors. While normal GM progenitors exhibited the expected synergistic response when KL was added to G-CSF + GM-CSF, CML GM progenitors had a minimal response; however, some synergism was restored by 10 nM PD173955. Normal erythroid progenitors require the synergistic interaction between KL and a saturating amount of erythropoietin (EPO, 1 unit) for optimal growth. In contrast, CML erythroid progenitors had up to 50% of optimal growth in KL alone, and, only a subthreshold amount of EPO (0.1 unit) was needed with KL to achieve 85% of the optimal response; these heightened growth responses were largely abrogated by 10 nM PD173955. Thus, direct evidence is provided that constitutively activated Bcr-Abl kinase pathways in primitive CML progenitors cooperate with single growth factors producing a heightened growth response, and, in so doing, disrupt the normally required synergistic interactions between KL and other cytokines to achieve activation and optimal growth of primitive progenitors. Coupled with our previous findings that a larger than normal proportion of CML primitive progenitors are at a later stage of maturation, we propose that this disruption of normal synergistic responses leads to increased progenitor recruitment into a committed pool by a process of accelerated maturation. PMID- 12556558 TI - Expression of Bcl-x(S) in Xenopus oocytes induces BH3-dependent and caspase dependent cytochrome c release and apoptosis. AB - The mechanism of action of pro-apoptotic proteins is difficult to study in vivo because of their death effect, which makes it problematic to obtain sufficient homogeneous experimental material for biochemical analysis. We show here that pro apoptotic genes expressed in Xenopus oocytes constitute a useful in vivo system for studying their mechanism of action. In the present study, we used this system to study the death effects of Bcl-x(S), a pro-apoptotic member of the Bcl-2 family. The results showed that expression of Bcl-x(S) in oocytes induces oocyte death by a caspase-dependent mechanism, which includes BH3-dependent cytochrome c release and is inhibited by co-expression of the anti-apoptotic proteins Bcl-2 and Bcl-x(L). The release of cytochrome c was found to be dependent on caspase activity. Bcl-x(S) was localized mainly in the mitochondria, and Bcl-x(S) transmembrane and BH3 domains were required for its apoptotic effect. These findings suggest that Bcl-x(S) induces apoptosis in Xenopus oocytes mainly by its presence in the mitochondria, where it induces BH3- and caspase-dependent release of cytochrome c, which leads to oocyte death. PMID- 12556559 TI - Cyclin G1 has growth inhibitory activity linked to the ARF-Mdm2-p53 and pRb tumor suppressor pathways. AB - Cyclin G1 is a p53-responsive gene that is induced in alternative reading frame (ARF)-arrested cells, yet its role in growth control is unclear. We tested its effects on growth and involvement in the ARF-Mdm2-p53 tumor suppressor pathway. We show that cyclin G1 interacts with ARF, Mdm2, and p53 in vitro and in vivo. At high levels, cyclin G1 induces a G(1)-phase arrest in mammalian cells that coincides with p53 activation. Conversely, lower levels of cyclin G1 lack intrinsic growth inhibitory effects yet potentiate ARF-mediated growth arrest. Notably, cyclin G1 is down-regulated by Mdm2 through proteasome-mediated degradation. These data suggest that cyclin G1 is a positive feedback regulator of p53 whose expression is restrained by Mdm2. Interestingly, growth inhibition by cyclin G1 does not require p53 but instead exhibits partial retinoblastoma protein (pRb) dependence. These findings reveal that cyclin G1 has growth inhibitory activity that is mechanistically linked to ARF-p53 and pRb tumor suppressor pathways. PMID- 12556560 TI - DNA replication-dependent nuclear dynamics of the Mre11 complex. AB - The Mre11 complex undergoes dramatic relocalization in the nuclei of gamma irradiated and replicating human cells. In this study, we examined Mre11 complex localization and chromatin association in synchronous cultures to examine the molecular determinants of relocalization. The data indicate that the complex is deposited on chromatin in an S phase-specific manner. Mre11 complex chromatin association in S phase was resistant to detergent extraction, in contrast to that in gamma-irradiated cells. The complex exhibits extensive colocalization with proliferating cell nuclear antigen throughout S phase, and chromatin loading is enhanced by replication fork stalling, suggesting that the replication fork is a site of Mre11 complex chromatin loading. This is supported by the observation that the complex localized to single-stranded DNA arising in hydroxyurea-treated cells. Although the Mre11 complex appears to function as a DNA damage sensor, limited colocalization with Brca1 or gamma-H2AX was observed, arguing that neither DNA damage nor gamma-H2AX is required for Mre11 complex chromatin loading. These data provide a potential molecular basis for promotion of sister chromatid association and recombination by the Mre11 complex as well as for ATM Mre11 complex-dependent activation of cell cycle checkpoints. PMID- 12556561 TI - Increased expression of epidermal growth factor receptor induces sequestration of extracellular signal-related kinases and selective attenuation of specific epidermal growth factor-mediated signal transduction pathways. AB - Increased expression of the epidermal growth factor receptor (EGFR) is common in cancer and correlates with neoplastic progression. Although the biology of this receptor has been the subject of intense investigation, surprisingly little is known about how increased expression of the wild-type EGFR affects downstream signal transduction in cells. We show that increasing the expression of the receptor results in dramatic shifts in signaling with attenuation of EGF-induced Ras, extracellular signal-related kinases (ERKs), and Akt activation, as well as amplification of STAT1 and STAT3 signaling. In this study, we focus on the mechanism of attenuated ERK signaling and present evidence suggesting that the mechanism of attenuated ERK signaling in EGFR-overexpressing cells is a sequestration of ERKs at the cell membrane in EGFR-containing complexes. Increased expression of the EGFR results in an aberrant localization of ERKs to the cell membrane. Furthermore, ERKs become associated with the EGFR in a physical complex in EGFR-overexpressing cells but not in control cells. The EGFR ERK association is detected in unstimulated cells or on exposure to a low concentration of EGF; under these conditions, ERK activation is minimal. Exposure of these cells to saturating concentrations of EGF results in a decreased membrane localization of ERKs, a concomitant dissociation of ERKs from the EGFR, and restores ERK activation. A similar association can be detected between the EGFR and MEK1 in receptor-overexpressing cells, suggesting that multiple components of the ERK signaling pathway may become trapped in complexes with the EGFR. These findings can be demonstrated in cells transfected to express high levels of the EGFR as well as in cancer cells which naturally overexpress the EGFR and, thus, may be representative of altered EGFR signaling in human cancer. PMID- 12556562 TI - The phosphoinositide 3-kinase/AKT1 pathway involvement in drug and all-trans retinoic acid resistance of leukemia cells. AB - Disruption of the apoptotic pathways may account for resistance to chemotherapy and treatment failures in human neoplastic disease. To further evaluate this issue, we isolated a HL-60 cell clone highly resistant to several drugs inducing apoptosis and to the differentiating chemical all-trans-retinoic acid (ATRA). The resistant clone displayed an activated phosphoinositide 3-kinase (PI3K)/AKT1 pathway, with levels of phosphatidylinositol (3,4,5) trisphosphate higher than the parental cells and increased levels of both Thr 308 and Ser 473 phosphorylated AKT1. In vitro AKT1 activity was elevated in resistant cells, whereas treatment of the resistant cell clone with two inhibitors of PI3K, wortmannin or Ly294002, strongly reduced phosphatidylinositol (3,4,5) trisphosphate levels and AKT1 activity. The inhibitors reversed resistance to drugs. Resistant cells overexpressing either dominant negative PI3K or dominant negative AKT1 became sensitive to drugs and ATRA. Conversely, if parental HL-60 cells were forced to overexpress an activated AKT1, they became resistant to apoptotic inducers and ATRA. There was a tight relationship between the activation of the PI3K/AKT1 axis and the expression of c-IAP1 and c-IAP2 proteins. Activation of the PI3K/AKT1 axis in resistant cells was dependent on enhanced tyrosine phosphorylation of the p85 regulatory subunit of PI3K, conceivably due to an autocrine insulin-like growth factor-I production. Our findings suggest that an up-regulation of the PI3K/AKT1 pathway might be one of the survival mechanisms responsible for the onset of resistance to chemotherapeutic and differentiating therapy in patients with acute leukemia. PMID- 12556563 TI - Expression and mechanistic analysis of a germacradienol synthase from Streptomyces coelicolor implicated in geosmin biosynthesis. AB - The PCR has been used to amplify a 2,181-bp ORF from Streptomyces coelicolor A3(2), designated SC9B1.20 (= SCO6073), encoding a protein of 726 amino acids and showing significant sequence similarity at the deduced amino acid level in both the N-terminal and C-terminal halves to the known sesquiterpene synthase pentalenene synthase. The full-length recombinant protein was expressed at high levels in Escherichia coli and shown to catalyze the Mg(2+)-dependent conversion of farnesyl diphosphate to the sesquiterpene alcohol (4S, 7R)-germacra-1 (10)E, 5E-diene-11-ol. The enzymatic cyclization had a k(cat) of 6.2 +/- 0.5 x 10(-3) s( 1) and a K(m) for farnesyl diphosphate of 62 +/- 8 nM. Expression of the N terminal (366 amino acids) domain of the SC9B1.20 protein also gave a fully functional cyclase which converted farnesyl diphosphate to the identical sesquiterpene alcohol with a slightly lower k(cat) of 3.2 +/- 0.4 x 10(-3) s(-1) and a twofold greater k(m) of 115 +/- 14 nM. By contrast, the expressed C terminal domain of SC9B1.20 had no farnesyl diphosphate cyclase activity. The formation of the germacradienol seems to be the committed step in the formation of geosmin, the characteristic odoriferous constituent of Streptomyces species. PMID- 12556564 TI - Engineering the metabolism of Escherichia coli W3110 for the conversion of sugar to redox-neutral and oxidized products: homoacetate production. AB - Microbial processes for commodity chemicals have focused on reduced products and anaerobic conditions where substrate loss to cell mass and CO(2) are minimal and product yields are high. To facilitate expansion into more oxidized chemicals, Escherichia coli W3110 was genetically engineered for acetate production by using an approach that combines attributes of fermentative and oxidative metabolism (rapid growth, external electron acceptor) into a single biocatalyst. The resulting strain (TC36) converted 333 mM glucose into 572 mM acetate, a product of equivalent oxidation state, in 18 h. With excess glucose, a maximum of 878 mM acetate was produced. Strain TC36 was constructed by sequentially assembling deletions that inactivated oxidative phosphorylation (deltaatpFH), disrupted the cyclic function of the tricarboxylic acid pathway (deltasucA), and eliminated native fermentation pathways (deltafocA-pflB deltafrdBC deltaldhA deltaadhE). These mutations minimized the loss of substrate carbon and the oxygen requirement for redox balance. Although TC36 produces only four ATPs per glucose, this strain grows well in mineral salts medium and has no auxotrophic requirement. Glycolytic flux in TC36 (0.3 micromol.min(-1).mg(-1) protein) was twice that of the parent. Higher flux was attributed to a deletion of membrane-coupling subunits in (F(1)F(0))H(+)-ATP synthase that inactivated ATP synthesis while retaining cytoplasmic F(1)-ATPase activity. The effectiveness of this deletion in stimulating flux provides further evidence for the importance of ATP supply and demand in the regulation of central metabolism. Derivatives of TC36 may prove useful for the commercial production of a variety of commodity chemicals. PMID- 12556565 TI - Identification of HIV-associated progressive multifocal leukoencephalopathy: magnetic resonance imaging and spectroscopy. PMID- 12556566 TI - Cognitive and motor functioning in gene carriers for Huntington's disease: a baseline study. AB - The aim of this study was to investigate the first changes in cognitive and motor functioning in Huntington's disease. Forty-six gene carriers, not clinically diagnosed for HD, were compared with 88 non-gene carriers. Gene carriers performed significantly worse on the Benton Visual Retention Test. This result was due to a minority of participants who had already developed cognitive impairment. Marginal differences appeared on the motor times of single reaction time measures after correction for motor signs. The findings are discussed in the context of inconsistencies in previous studies and underscore the need for longitudinal research. PMID- 12556567 TI - A review of cholinergic agents in the treatment of neurobehavioral deficits following traumatic brain injury. AB - Although traumatic brain injury (TBI) frequently results in significant handicap, empirical investigations of pharmacological treatment of the neurobehavioral sequelae of TBI are rare. This review presents evidence that supports hypotheses of a cholinergic mechanism underlying some neurobehavioral sequelae of TBI, as well as a critical review of the preliminary evidence supporting the efficacy of cholinergic agents in TBI. Despite numerous methodological limitations, preliminary evidence exists for the efficacy of cholinergic agents in ameliorating attention and memory deficits following TBI. The authors highlight the need for large, randomized, double-blind, placebo-controlled trials that include a broad range of cognitive and behavioral outcome measures. PMID- 12556568 TI - Comparison of seizure duration, ictal EEG, and cognitive effects of ketamine and methohexital anesthesia with ECT. AB - The authors retrospectively compared the seizure duration, ictal EEG, and cognitive side effects of ketamine and methohexital anesthesia with ECT. This comparison was carried out with data from consecutive index ECT treatments that occurred immediately before and after a switch from methohexital to ketamine in 36 patients. Ketamine was well tolerated and prolonged seizure duration overall, but particularly in those who had a seizure duration shorter than 25 seconds with methohexital at the maximum available stimulus intensity. Ketamine also increased midictal EEG slow-wave amplitude. Thus, a switch to ketamine may be useful when it is difficult to elicit a robust seizure. Faster post-treatment reorientation with ketamine may suggest a lower level of associated cognitive side effects. PMID- 12556569 TI - Regional cerebral blood flow changes during visually induced subjective sadness in healthy elderly persons. AB - This study examined regional cerebral blood flow (rCBF) changes associated with visually induced sad affect in healthy elderly persons. Subjects viewed sadness laden, happiness-laden, and emotionally neutral image sets while rCBF was recorded using [(15)O] water PET. The sad image set included human faces and scenery/objects ("scenes"). To control for secondary sensory processing, the neutral and happy comparison sets included exclusively either human faces or scenes. During the sad condition, the ventral prefrontal and temporal cortices were more active compared with happy and neutral scenes conditions and the thalamus was more active compared with happy and neutral faces conditions. Ventral prefrontal cortex and thalamus were associated with processing of sad visual stimuli, whether compared with neutral or happy stimuli. The specific findings associated with sad affect were contingent on the comparison stimuli content (scenes or human faces), not affect (i.e., comparison with neutral or happy conditions). PMID- 12556571 TI - No evidence of a homogeneous frontal neuropsychological profile in a sample of schizophrenic subjects. AB - The authors obtained a frontal functions profile for 81 schizophrenic patients using six neuropsychological tests that evaluate the dorsolateral prefrontal cortex functions, i.e., working memory, executive functions, and strategic performance. They then analyzed the test performances with a cluster analysis, which produced a four-cluster solution. The results support the hypothesis that neuropsychological dysfunctions in schizophrenia are heterogeneous. The performances on many of the neuropsychological tests were also strongly correlated with verbal and nonverbal IQ, as measured by the Wechsler Adult Intelligence Scale-Revised. PMID- 12556570 TI - Are late-onset schizophrenia spectrum disorders neurodegenerative conditions? Annual rates of change on two dementia measures. AB - Some investigators assert that emergence of schizophrenia spectrum disorders in mid- to late life (LOSD, i.e., onset after age 45) reflects neurodegenerative processes. The authors examined 1- and 2-year changes among 37 outpatients with LOSDs on the Mini-Mental State Examination (MMSE) and Mattis Dementia Rating Scale, compared with 69 patients having earlier-onset schizophrenia spectrum disorders (EOSD), 67 having Alzheimer's disease (AD) with psychosis, 72 having AD with baseline MMSE scores >/=24, and 56 normal comparison subjects (NCs). Cognitive changes among LOSD patients were similar to those in EOSD patients and NCs, whereas the AD groups had greater declines. Results support viewing LOSDs as static encephalopathies. PMID- 12556572 TI - Mild procedural learning disturbances in neuroleptic-naive patients with schizophrenia. AB - This study investigated whether drug-naive patients with schizophrenia present procedural learning disturbances, given that this function is known to involve the striatum and that structural abnormalities of the striatum have been described in these patients. Eleven neuroleptic-free schizophrenic patients were compared with 11 healthy control subjects on a visuomotor procedural learning task (the Mirror Drawing Test). Average performance and learning rate did not differ between the two groups. However, schizophrenia patients were lower in learning smoothness (the consistency with which performance improves from trial to trial), which would indicate difficulty in benefiting from past experience to perform better on a current task. Because low smoothness has recently been found to be associated with striatal dysfunction, these results might indicate a premature dysfunction of the striatum in young drug-naive patients with schizophrenia. However, this dysfunction is not severe enough to prevent learning. PMID- 12556573 TI - Low serum folate levels as a risk factor for depressive mood in patients with chronic epilepsy. AB - This study takes into consideration whether low serum folate levels may contribute to depressive mood in patients with chronic epilepsy. The serum folate levels and the score on the Self-Rating Depression Scale (SDS) were examined in 46 patients with chronic epilepsy. Patients with a score indicating at least minor depression on the SDS had a significantly lower serum folate level than patients with a normal score on SDS. There was a significant negative correlation between the serum folate levels and the SDS score. A serum folate level below 7.5 ng/ml was significantly associated with a pathological score on SDS. Because a serum folate level of 7.5 ng/ml is in the normal range for many laboratories, further studies using total plasma homocysteine as a sensitive measure of functional folate deficiency are required to elucidate the impact of folate metabolism on depressive mood in patients with chronic epilepsy. PMID- 12556574 TI - Patterns of change in the treatment of psychiatric symptoms in patients with probable Alzheimer's disease from 1983 to 2000. AB - The authors examined the pattern of use of psychiatric medication as prescribed by community physicians in 1,155 patients with probable Alzheimer's disease (AD) referred to the Alzheimer's Disease Research Center of Pittsburgh between April 1983 and July 2000. The use of antidepressants and of sedatives, hypnotics, and anxiolytics (SHA) increased over time, while the use of antipsychotics decreased. The increased use of antidepressants and decreased use of antipsychotics may reflect the growing evidence that newer antidepressants (e.g., selective serotonin reuptake inhibitors) can be used to treat not only mood-related disorders, but also abnormal behavior (e.g., aggression, agitation) and sleep disorders in AD. Although the use of SHA has a proven deleterious effect on patients with AD, their use has increased over the past two decades. PMID- 12556575 TI - The contribution of somatic symptoms to the diagnosis of depressive disorder in Parkinson's disease: a discriminant analytic approach. AB - This study assessed the sensitivity of individual depressive symptoms and their relative contribution to the diagnosis of depressive disorder in patients with Parkinson's disease. The Structured Clinical Interview for DSM-IV Depression and the Hamilton and Montgomery-Asberg depression rating scales (Ham-D, MADRS) were administered to 149 consecutive nondemented patients. The contribution of the individual items of these scales to the diagnosis of "depressive disorder" was calculated by discriminant analysis. The discriminant models based on the Ham-D and MADRS scores were both highly significant. Nonsomatic core symptoms of depression had the highest correlation coefficient. Somatic items had mostly low correlation coefficients, with the exception of reduced appetite and early morning wakening (late insomnia). Nonsomatic symptoms of depression appear to be the most important for distinguishing between depressed and nondepressed patients with Parkinson's disease, along with reduced appetite and early morning awakening. PMID- 12556576 TI - Influence of the apolipoprotein E type 4 allele on cerebral glucose metabolism in Alzheimer's disease patients. AB - It is still unclear whether apolipoprotein E epsilon 4 (APOE E4) influences the cerebral glucose metabolism abnormalities found in Alzheimer's disease (AD), although APOE E4 is a well-known risk factor for AD. [(18)F]Fluorodeoxyglucose PET was conducted in patients with very mild (n=17), mild (n=27), and moderate-to severe (n=19) AD. The presence of APOE E4 was associated with greater reduction of glucose metabolism in the left inferior temporal region in very mild AD but neither in mild nor in moderate-to-severe AD. These findings favor the hypothesis that APOE E4 is related mainly to the development of AD, not to its progression. PMID- 12556578 TI - Progress of memory function after radiation therapy in patients with nasopharyngeal carcinoma. AB - The authors evaluated the late neurocognitive profile and progress of memory functions in 60 patients with primary nasopharyngeal carcinoma who had been treated with radiation therapy for more than 2 years. Forty had imaging evidence of temporal lobe injury (TLI-positive), and 20 did not (TLI-negative). The patients and 19 healthy control subjects, matched with the patients for age and educational level, underwent extensive memory assessments. Seventeen patients (10 TLI-positive, 7 TLI-negative) were reassessed after an average of 28 months for progress of memory functions. The patient groups performed significantly worse on most memory tests compared with the normal subjects. Patients with memory complaints had lower scores for verbal memory than those without such complaints. The TLI-positive and TLI-negative groups did not differ significantly from each other in cognitive performance. At follow-up, visual memory performance had deteriorated, while verbal memory remained more stable. PMID- 12556577 TI - Cognitive deficits in patients with hepatic cirrhosis and in liver transplant recipients. AB - This study assessed cognitive deficit in patients diagnosed with different stages of hepatic cirrhosis and in liver transplant recipients. A short protocol consisting of several psychometric tests was used. Cirrhotic patients showed a degree of mental impairment in all the functions studied. The severity of the deficit was related to the degree of hepatic dysfunction. In contrast, liver transplant recipients presented only a slightly altered cerebral dysfunction in comparison to the control group. Their cognitive capacity was slightly better than that of patients with asymptomatic cirrhosis. PMID- 12556579 TI - Effects of depressed mood on objective and subjective measures of attention. AB - People with depression report frequent cognitive failures, but objective measures of cognition show mixed results. Some studies show impairment on effortful tasks. The relationship between subjective and objective cognitive failures was studied in 102 "depressed" or "nondepressed" UK servicemen, grouped by Beck Depression Inventory scores with a cutoff score of 10. Participants were administered cognitive tests including the Sustained Attention to Response Task (SART), a laboratory measure of vigilance that has revealed increased attentional lapses in traumatic brain injury patients. The depressed men made more errors on SART than the nondepressed men (P=0.012) but reported much higher incidences of cognitive failures on a standardized questionnaire (P=0.0001). The depressed men's SART reaction times slowed following an error, a pattern different from that of brain injured subjects. Nonclinical depressed subjects may respond "catastrophically" to errors, heightening the subjective sense of failure and contributing to the strong relationship between subjectively reported cognitive failures and depression. PMID- 12556580 TI - Impaired olfactory identification in Asperger's syndrome. AB - The authors measured odor detection threshold and odor identification in 12 males with Asperger's syndrome and 12 matched control subjects. Relative to control subjects, Asperger's syndrome subjects were not impaired at odor detection but were significantly impaired at olfactory identification. PMID- 12556581 TI - Neurodegenerative disorders: George Huntington's description of hereditary chorea. PMID- 12556582 TI - On chorea. George Huntington, M.D. PMID- 12556583 TI - Thalamic acalculia. PMID- 12556584 TI - Heroin and malignant coprolalia in Tourette's syndrome. PMID- 12556585 TI - On Bresch's glossary of eponyms. PMID- 12556587 TI - Atypical antipsychotic use among acute schizophrenic inpatients. PMID- 12556589 TI - Large data sets can be dangerous! PMID- 12556591 TI - Economic grand rounds: psychopharmacosocioeconomics and the global burden of disease. PMID- 12556592 TI - Psychopharmacology: number needed to treat: an underused measure of treatment effect. PMID- 12556593 TI - Practical psychotherapy: integrated psychotherapy. PMID- 12556594 TI - Personal accounts: mental illness, motherhood, and me. PMID- 12556595 TI - Datapoints: U.S. rates of self-inflicted injuries and suicide, 1992-1999. PMID- 12556596 TI - Alcohol & drug abuse: principles of money management as a therapy for addiction. PMID- 12556597 TI - Best practices: racial and ethnic effects on antipsychotic prescribing practices in a community mental health center. PMID- 12556598 TI - A comparison of the performance of for-profit and nonprofit U.S. psychiatric inpatient care providers since 1980. AB - OBJECTIVE: The authors synthesized evidence from a systematic review of the literature reporting substantiated performance differences between private for profit and private nonprofit psychiatric inpatient care providers in the United States since 1980. They also compared reported differences in performance between nonprofit and for-profit inpatient psychiatric care providers with reported differences between nonprofit and for-profit providers of other types of health care. METHODS: Studies were located by means of computerized bibliographic searches and follow-up searches of studies cited in the articles located in the computerized search. The analysis included peer-reviewed studies that compared the performance of for-profit and nonprofit health service providers, including inpatient psychiatric services, in the areas of access, quality, cost-efficiency, and amount of charity care on the basis of quantitative data collected after 1980. The studies were classified in one of three categories according to the study conclusion: for-profit superiority, nonprofit superiority, or no difference or mixed results. RESULTS: Almost all studies (with one exception) found that the nonprofit psychiatric providers performed as well as or better than their for profit counterparts. The proportion of studies reporting performance superiority of nonprofit versus for-profit psychiatric inpatient providers was greater than the proportion of studies reporting the same conclusion for providers of all other types of health care taken together. CONCLUSIONS: On the basis of data collected since 1980, nonprofit psychiatric inpatient care providers in the United States had superior performance on access, quality, cost-efficiency, and amount of charity care, compared with for-profit providers. Caution is warranted in pursuing public policies that permit or encourage the replacement of nonprofit psychiatric inpatient care providers with for-profit providers of these services. PMID- 12556599 TI - Medicaid expenditures on behavioral health care. AB - OBJECTIVE: The authors reviewed studies of Medicaid spending on mental health and substance abuse services. METHODS: Studies were identified through a search of MEDLINE and bibliographies of known articles on mental health and substance abuse spending and by searching Web sites of or contacting key government and private organizations. Of 448 studies identified, the 14 that included Medicaid expenditure percentages for 1984 or later were compared. RESULTS AND CONCLUSIONS: The most comprehensive studies of such spending suggest that between 9.3 and 13 percent of all Medicaid dollars are spent on behavioral health services. The most comprehensive estimates came from claims-based studies or studies based on the National Health Accounts. Studies based on provider or consumer surveys missed large portions of Medicaid spending. Policy makers need to ensure that they use the most accurate data to track mental health and substance abuse spending, an important part of total Medicaid expenditures. PMID- 12556600 TI - Strategies to decrease costs of prescribing selective serotonin reuptake inhibitors at a VA Medical Center. AB - OBJECTIVE: The authors examined the efficacy of a multifaceted intervention designed to contain the cost of prescribing selective serotonin reuptake inhibitors (SSRIs) to inpatients and outpatients served by a Veterans Affairs (VA) medical center. METHODS: Elements of the intervention included identification of a preferred agent, tablet splitting, education and feedback for prescribers, and an electronic record and ordering system to facilitate changes in prescriber behaviors. VA databases were searched for information on use and costs of antidepressants. RESULTS: Over 35 months the number of patients treated with SSRIs and the amount spent on SSRIs increased. However, the mean monthly cost per patient decreased from $57.12 to $42.19. The projected cost savings over the 35 months was approximately $700,000; one-fourth of the savings was due to tablet splitting and three-fourths to changes in the proportions of the various SSRIs prescribed. A survey of the top 75 antidepressant prescribers showed that after the educational interventions, 91 percent were aware that citalopram was the medical center's preferred antidepressant, and 59 percent identified it as their own preferred first-line treatment. DISCUSSION AND CONCLUSIONS: The results suggest that multifaceted interventions can influence antidepressant costs through provider education and changes in pharmacy and computerized information processes, resulting in substantial cost savings for institutions. PMID- 12556601 TI - Health status, service use, and costs among veterans receiving outreach services in jail or community settings. AB - OBJECTIVE: This study compared client characteristics, service use, and health care costs of two groups of veterans who were contacted by outreach workers: a group of veterans who were contacted while incarcerated at the Los Angeles jail and a group of homeless veterans who were contacted in community settings. METHODS: Between May 1, 1997, and October 1, 1999, a total of 1,676 veterans who were in jail and 6,560 community homeless veterans were assessed through a structured intake procedure that documented their demographic, clinical, and social adjustment characteristics. Data on the use and costs of health services during the year after outreach contact were obtained from national databases of the Department of Veterans Affairs (VA). Chi square and t tests were used for statistical comparisons. RESULTS: The veterans who were contacted in jail obtained higher scores on several measures of social stability (marital status and homelessness status) but had higher rates of unemployment. They had fewer medical problems but higher levels of psychiatric and substance use problems, although the rate of current substance use was lower among these veterans than among the community homeless veterans. One-year service access for the jailed veterans was half that of the community homeless veterans. No differences were observed in the intensity of use of mental health services among those who used services, but the jailed outreach clients used fewer residential, medical, and surgical services. Total health care expenditures for the veterans who received outreach contact in jail were $2,318 less, or 30 percent less, than for those who were contacted through community outreach. CONCLUSIONS: Specialized outreach services appear to be modestly effective in linking veterans who become incarcerated with VA health care services. Although it is clinically challenging to link this group with services, the fact that the rate of current substance use is lower during incarceration may provide a window of opportunity for developing linkages between inmates and community rehabilitative services. PMID- 12556602 TI - Recognition of depression among elderly recipients of home care services. AB - OBJECTIVE: Depression among older home care patients is prevalent but undertreated. The purpose of this study was to investigate the ability of home health nurses to correctly identify depression among older patients and to describe nurse and patient characteristics associated with more accurate assessment of depression. METHODS: Forty-two nurses were surveyed about the presence of depressive symptoms among patients who had been evaluated independently for depressive disorders by research staff using the Structured Clinical Interview for Axis I DSM-IV Disorders. A sample of newly admitted home health care patients who were aged 65 years or older was randomly selected for this evaluation on a weekly basis from December 1997 to December 1999. RESULTS: Of 403 patients who were evaluated, 97 (24 percent) were found to have either major depression (64 patients) or minor depression (33 patients). The nurses correctly identified depression among 44 of the 97 patients who were depressed (sensitivity of 45.4) and 230 of the 306 patients who were not depressed (specificity of 75.2). The kappa coefficient measuring overall agreement between the nurses' assessment and the diagnosis of depression was.19. Nurses who had more geriatric nursing experience were more likely to correctly identify depression. CONCLUSIONS: Home health nurses have difficulty making accurate assessments of depression among older home care patients. Inaccuracy in assessment of depression by home health nurses is a significant barrier to treatment in this elderly homebound population. PMID- 12556603 TI - Screening for substance abuse and psychiatric disorders among women patients in a VA Health Care System. AB - OBJECTIVE: This study of women Veterans Affairs (VA) health care patients screened for the prevalence of past-year smoking, hazardous and problem drinking, other drug abuse, and psychiatric disorders. METHODS: A survey was mailed to women veterans who had received care from VA Puget Sound Health Care System between October 1, 1996, and January 1, 1998. Screening measures included questions about cigarettes; questions from the Alcohol Use Disorders Identification Test about consumption (hazardous drinking); the TWEAK test (problem drinking); a drug abuse screen; the Patient Health Questionnaire (psychiatric conditions); and the PTSD (posttraumatic stress disorder) Checklist. RESULTS: Of eligible patients, 1,257 (65 percent) returned surveys with complete substance use data. Patients reported a relatively high rate of past-year smoking (29.1 percent) and hazardous drinking, problem drinking, or both (31.1 percent). The rate of past-year drug use was much lower (4.9 percent). Younger age was strongly associated with greater substance abuse: 59 percent of women under age 35 screened positive for smoking, hazardous or problem drinking, or drug abuse. Screening positive for a psychiatric condition (N=504) was also associated with substance abuse: The rate of past-year drug abuse among women screening positive for a psychiatric condition (9.7 percent) was double the rate for the entire sample. Of the women who screened positive for depression, PTSD, eating disorders, or panic disorders, 57 percent screened positive for substance abuse (including smoking). CONCLUSIONS: Substance abuse is common among women VA patients and is associated with younger age and with screening positive for other psychiatric conditions. Providers are expected to follow up on positive screening tests, and these data indicate substantial provider burden. PMID- 12556604 TI - Treatment of depression among impoverished primary care patients from ethnic minority groups. AB - OBJECTIVE: The aim of this study was to determine whether supplementing traditional cognitive-behavioral therapy for depression with clinical case management would reduce the rate of dropout from care and improve outcomes for ethnically diverse, impoverished medical outpatients. METHODS: The study was a randomized trial that compared cognitive-behavioral group psychotherapy alone (N=103) with the same therapy supplemented by clinical case management (N=96). RESULTS: The patients who received supplemental case management had lower dropout rates than those who received cognitive-behavioral group therapy alone. Supplemental case management was associated with greater improvement in symptoms and functioning than cognitive-behavioral therapy alone for patients whose first language was Spanish (N=77) but was less effective for those whose first language was English (N=122). CONCLUSIONS: Supplemental case management improves retention in traditional mental health outpatient care and can improve outcomes for Spanish speaking patients. PMID- 12556605 TI - Patterns of service use among persons with schizophrenia and other psychotic disorders. AB - OBJECTIVE: This study assessed 12-month service use patterns among people with psychotic disorders and sought to identify determinants of service use. METHODS: As part of a large two-phase Australian study of psychotic disorders, structured interviews were conducted with a stratified random sample of adults who screened positive for psychosis. Demographic characteristics, social functioning, symptoms, mental health diagnoses, and use of psychiatric and nonpsychiatric services were assessed. Data were analyzed for 858 persons who had an ICD-10 diagnosis of a psychotic disorder and who had been hospitalized for less than six months during the previous year. RESULTS: People with psychotic disorders had high levels of use of health services, both in absolute terms and relative to people with nonpsychotic disorders. Those with psychotic disorders were estimated to have an average of one contact with health services per week. Use of psychiatric inpatient services was associated with parenthood, higher symptom levels, recent attempts at suicide or self-harm, personal disability, medication status, and frequency of alcohol consumption. Services provided by general practitioners (family physicians) were more likely to be obtained by older people, women, people with greater availability of friends, those with fewer negative symptoms, and those whose service needs were unmet by other sources. People who were high users of health services also reported having more contact with a range of non-health agencies. CONCLUSIONS: The predictors of service use accounted for small proportions of the variance in overall use of health services. The role of general practitioners in providing and monitoring treatment programs and other psychosocial interventions needs to be acknowledged and enhanced. PMID- 12556606 TI - Academic psychiatry and health care reform: strategic initiatives for sustaining the clinical mission. AB - Health care reform has posed special challenges for departments of psychiatry in academic medical centers. This report describes one department's strategic responses to a marketplace with high penetration by managed care and provides examples of the kinds of faculty concerns that can arise when major departmental reorganizations are attempted. The department's successful adaptation to a radically altered professional environment is attributed to the following five initiatives: vertical integration and diversification of clinical programs, service line management, outcomes measurement, regional network development, and institutional managed care partnerships Although the authors did not design their adaptive efforts as a research study, they offer objective data to support their conclusion that the viability of their overall clinical enterprise has been sustained despite an external environment inhospitable to academic psychiatry. PMID- 12556607 TI - Characteristics of patients presenting for emergency psychiatric assessment at an English hospital. AB - OBJECTIVE: The needs and characteristics of patients who are referred for psychiatric emergency services vary by the source of referral. Such differences have wider implications for the functioning of the mental health care system as a whole. This study compared three groups of patients in a two-month cohort of 189 patients who were referred for emergency psychiatric assessment at a hospital in England: those who were referred by general practitioners (family physicians), those who were receiving specialist services from community mental health teams, and those who arrived at the hospital from the broader community. METHODS: The three groups were compared on demographic characteristics, clinical and service use variables, risk to self or others, factors that contributed to the emergency presentation, and ratings on standardized scales of functioning. RESULTS: The patients who were receiving specialist services from community mental health teams had high rates of psychosis, often relapsed, and had a history of contact with a psychiatrist. These patients were the most likely to be admitted to the hospital after emergency assessment. The patients who had been referred by general practitioners tended to have fewer indicators of social problems and were more likely to be experiencing a new episode of mental illness. Their referral to the emergency department was most likely to be deemed inappropriate by emergency department clinicians. The patients who came from the broader community were more likely to be male and to exhibit self-harming behavior, substance misuse, and behavioral difficulties. CONCLUSIONS: The rate of emergency referral is one indicator of the functioning of the service system as a whole. Improvements to the system should include better access to community mental health team services and a greater capacity of the primary care system to manage mental health crises. Services need to be developed that are acceptable to male patients who are experiencing social and behavioral problems. PMID- 12556608 TI - Prescribers' nonadherence to treatment guidelines for schizophrenia when prescribing neuroleptics. AB - Treatment guidelines are frequently not followed. The authors examined the neuroleptic prescribing practices of psychiatrists responsible for 47 patients with schizophrenia who were being treated at health centers of the Department of Veterans Affairs. For 22 of these patients, a medication change was indicated by guidelines previously endorsed by the prescriber; for 21 of these 22 patients, prescribers indicated that a change in neuroleptic would not be attempted; for 15 patients (71 percent), the reason given was the patient's refusal to change medication or the patient's noncompliance with medication treatment. The results suggest that patients' agreement with treatment guidelines should be taken into account in the evaluation of prescribers' use of such guidelines. PMID- 12556610 TI - The medical expertise of psychiatrists. PMID- 12556609 TI - Prevalence of psychiatric disorders among bereaved survivors of a disastrous earthquake in taiwan. AB - This study investigated the prevalence of psychiatric disorders and risk factors for posttraumatic stress disorder (PTSD) and major depressive disorder among bereaved survivors of a severe earthquake in Taiwan. A total of 120 survivors received a psychiatric interview conducted by board-certified psychiatrists. The two most prevalent disorders were PTSD (37 percent) and major depressive disorder (16 percent). The risk factors for PTSD were psychosocial stressors and initial feelings of guilt. The risk factor for major depressive disorder was female gender. Only 25 percent of the survivors with PTSD and 26 percent of those with major depressive disorder sought help at primary care clinics. These results indicate a need to develop an effective outreach strategy for dealing with psychiatric disorders among disaster victims. PMID- 12556611 TI - Quick Reference Guides for APA Practice Guidelines. PMID- 12556612 TI - "Iatrogenic Malingering" in VA substance abuse treatment. PMID- 12556613 TI - Depression and quality of life among depressed Brazilian outpatients. PMID- 12556615 TI - Post-cardiac catheterization access site complications and low-molecular -weight heparin following cardiac catheterization. AB - The low molecular weight heparin enoxaparin is often administered to patients on long-term anticoagulation regimens who temporarily discontinue warfarin prior to undergoing invasive procedures. The clinical outcome of all enoxaparin-treated patients who underwent cardiac catheterization or coronary artery interventional procedures (n = 119) was evaluated. A total of 5 patients (4.2%) requiring anticoagulation (3 with chronic atrial fibrillation and 2 with ventricular thrombi) developed severe late enoxaparin-associated hemorrhagic or vascular complications at the femoral arterial puncture site between 3 and 11 days post procedure. Complications included development of femoral arterial pseudoaneurysm (n = 3), hypotension (systolic blood pressure < 90 mmHg) (n = 2), acute decrease in hemoglobin levels to < 8.5 mg/dl (n = 4) and cardiac arrest (n = 1). In patients receiving standard dose enoxaparin after percutaneous invasive cardiac procedures, there is the potential for delayed and severe access site hemorrhagic and vascular complications. PMID- 12556616 TI - Long-term warfarin and percutaneous intervention. PMID- 12556617 TI - Successful aspiration of occlusive coronary thrombus with intracoronary aspiration using the export catheter. PMID- 12556618 TI - A comparison of arterial closure devices to manual compression in liver transplantation candidates undergoing coronary angiography. AB - Whether arterial closure devices can be used safely in a coagulopathic population undergoing cardiac catheterization and at high risk for groin complications, such as liver transplant candidates, is unknown. In this prospective, non-randomized consecutive series of 80 liver transplant candidates undergoing coronary angiography, manual compression and arterial closure devices were compared. Ilio femoral angiography was performed to determine suitability for use of the closure device. Bleeding and vascular complications were recorded along with time to ambulation. Arterial closure devices were used in 31 patients (39%), whereas manual compression was used in 49 patients (50 procedures) (61%). There were no significant differences between the two groups with respect to age, sex, cardiac risk factors, peripheral vascular disease, baseline platelet count or baseline INR. There were 10 total vascular complications out of 50 procedures (20%) in the manual compression group compared to 2 vascular complications out of 31 procedures in the arterial closure device group (6%; p = 0.12). The time to ambulation was significantly less in the group receiving arterial closure devices versus manual compression (4.2 1.8 hours versus 6.6 3.7 hours, respectively; p = 0.0003). In coagulopathic patients at higher risk for groin complications, arterial closure devices can be safely used and decrease time to ambulation compared to manual compression. PMID- 12556619 TI - Contemporary view of the acute coronary syndromes. PMID- 12556620 TI - Selective coronary artery fistula embolization with hystoacryl during percutaneous coronary angioplasty. AB - The percutaneous approach to coronary artery fistula (CAF) is diverse; coils, detachable balloons and double-umbrella devices can be used. Selective embolization with hystoacryl is a novel technique that has been successfully used in patients with intracerebral arteriovenous fistulae. Hystoacryl is a resin, which on contact with the bloodstream immediately solidifies, causing complete luminal obliteration of the fistulous pathway. In the present report, we describe two cases of CAF draining into the pulmonary artery. These two cases were successfully treated with hystoacryl embolization of the fistulous pathway, during the same procedure of coronary angioplasty for localized atherosclerotic lesions in major epicardial vessels. PMID- 12556621 TI - Treatment of iatrogenic aortic dissection by percutaneous stent placement. AB - During elective cardiac catheterization via the right femoral artery, difficulty was encountered advancing the guidewire beyond the aortic arch and the procedure abandoned. The patient later developed back pain and hypotension. A contrast enhanced spiral computed tomography scan confirmed aortic dissection, extending from an entry point in the external iliac artery to the arch. We hypothesized that the entry site was responsible for filling the dissection in the thoraco abdominal component. A stent was placed in the iliac artery, occluding the entry point. This is the first report of aortic dissection, a rare complication of angiography, managed by percutaneous stenting of the entry point. PMID- 12556622 TI - Stenting through a portacath for totally occluded superior vena cava in a case of non-Hodgkin s lymphoma. AB - Superior vena cava (SVC) syndrome often presents with slowly progressive symptoms worsening over weeks or may cause abrupt symptoms and constitute a true medical emergency. Percutaneous intervention is the treatment of choice. We report a case of SVC stenting in a middle-aged woman with SVC obstruction secondary to portacath insertion for chemotherapy. PMID- 12556623 TI - Aortocoronary dissection complicating a percutaneous coronary intervention. AB - Aortocoronary dissection complicating diagnostic angiography or percutaneous coronary intervention is a rare occurrence. We present a case of aortocoronary dissection complicating a right coronary artery intervention. After closing the entry port with intracoronary stenting, the patient was managed conservatively. Serial computed tomography scans demonstrated spontaneous resolution of the dissection. PMID- 12556624 TI - Intracardiac echocardiography and transcranial Doppler ultrasound to guide closure of patent foramen ovale. AB - Intracardiac echocardiography is the newest imaging technology permitting whole heart evaluation from a right-sided catheter position and contrast transcranial Doppler is an ideal method to noninvasively quantify right-to-left cardiac shunt in real time, by the detection of bubble passage across the brain arteries. The combination of these two methods enables us to take advantage of online therapeutic information for successful deployment of the Amplatzer PFO Occluder in a 63-year-old man with severe chronic obstructive pulmonary disease, avoiding the need for transesophageal echocardiography and general anaesthesia or sedation. PMID- 12556625 TI - Obliteration of a competitive forward flow from the ventricle after a bidirectional cavopulmonary shunt with an Amplatzer duct occluder. AB - A 7-month-old boy who was status post-bidirectional Glenn shunt implantation with residual ventricular flow to the pulmonary arteries (PA) presented with massive edema of the head and systemic desaturation soon after surgery. After test occlusion, an Amplatzer duct occluder was implanted at the PA banding site with subsequent decrease in the PA pressure, resolution of the edema and extubation. PMID- 12556626 TI - Platelet GP IIb/IIIa inhibition in unstable angina and non-ST segment elevation MI: application of the ACC/AHA guidelines. PMID- 12556627 TI - [Results of the treatment of kidney and ureteral stones by the method of extracorpored shock wave lithotripsy at the Klaipeda Hospital]. AB - OBJECTIVE: review the cases, which are cured by extracorpored shock wave lithotripsy using the device of third generation "Lithostar Multiline" (Germany). There are data of 603 patients treated by extracorpored shock wave lithotripsy. Almost a thousand (979) renal stones' fragmentations were performed for 271 (45%) patients. The analysis had showed: a full success in 225 (83%) patients, partial in 28 (12.4%) ones and there wasn't any fragmentation in 18 (6.6%) of cases. Extracorpored wave lithotripsy of ureteral stones was performed in 364 (60.4%) cases. Full success was in 319 (87.6%) cases; partial fragmentation in 25 (6.9%) cases and there wasn't any effect in 24 (8%) cases. PMID- 12556628 TI - [Review and results of our first ureteroscopy experience at the Klaipeda Hospital]. AB - OBJECTIVE: To estimate an experience of our first ureteroscopic procedures. MATERIAL AND METHODS: The data of 65 patients, who underwent ureteroscopy in Klaipeda Hospital during period 2000 August - 2002 February were analyzed. The indications were: obstructive ureteral stones after not successful extracorporeal lithotripsy, rigid ureteral stones, radionegative ureteral stones and ureteral strictures. The procedures were performed mostly in general, less in spinal anesthesia. The operations were completed on average within 120 minutes. The whole procedure was performed in double visual and radiologic correction. In 42.3 percent of cases there was performed only lithextraction, in other 42.3 percent of cases pneumatic lithotripsy and lithextraction, the remainder underwent for fixing with Dormia basket, pneumatic lithtripsy and lithextraction. RESULTS: There were 65 ureteroscopies performed, of them 59 cases for the reason of ureterolithiasis. The overall success rate for ureterolithiasis was 92.3 percent. Two cases were transformed in open operation. All patients received postoperative stenting, which lasted from 3-4 days to several weeks. The required hospitalization after ureteroscopy was on average 4.7 days. No serious operative and postoperative complications were noticed. CONCLUSIONS: Ureteroscopy minimally invasive, effective and comparatively safe method of treatment for complicated ureterolithiasis. This may be second - line treatment of ureteral stones after extracorporeal lithotripsy and even the choice of first line of treatment in some cases. An experience of surgeon is very definitive. PMID- 12556629 TI - [The significance of ultrasound examination in the assessment of kidney tumors: TNM criteria and stages]. AB - Ultrasound (US) examination was performed for 144 patients, operated with kidney tumors. For 13 (9.03%) patients 1 disease stage was determined. For 12 patients first disease stage was certified during operation and by histological examination after operation. For 1 patient, during histological examination, II stage was certified. II stage histologically was certified for 62 patients. It is 43.06% of all examined patients. By US examination II stage was suspected for 72 patients (50.0%), but after operation and histological examination, for 10 patients was certified III stage. II stage by US examination, was determined for 58 (40.28%) patients, but after operation and histological examination for 5 patients was certified II stage. PMID- 12556630 TI - [Renal trauma in children]. AB - In 1987 and 1999, fifty five children were treated with a renal injury after blunt abdominal trauma at the Department of Pediatric Surgery of Vilnius University Children Hospital. There were 40 boys and 15 girls. Using a G.S. Nicolaisen and J.W. McAninch (1985) classification of injury staging, 23 (41.8%) patients were ascertained as having type I, 16 (29.1%) - type II, 15 (27.3%) - type III, 1 (1.8%) - type IV. Conservative methods were used for treatment in 91% children and 4 (7.3%) had indication for operative treatment: 2 of these children underwent nephrectomy, in 2 cases kidneys were sutured. PMID- 12556631 TI - [Hydronephrosis in pregnancy: importance of urologic actions and their volume]. AB - In the present study, the data from 216 women with a complicated hydronephrosis of pregnancy who were treated at the Urology Department of the Vilnius City University Hospital between 1992 and 2001 have been subjected by us to a retrospective analysis and to an evaluation of the need for an interventional treatment and of the volume of the latter in the light of the special literature of the last decade of the XX(th) century. Both urologic observation and urologic management are required in the pregnancy hydronephrosis complicated by strong flank pains due to an acute dilatation of the upper urinary tract or by urolithiasis or pyelonephritis. An ultrasonographic investigation is the diagnostic mean of choice. The conservative management (hydration, spasmoanalgetics and antibiotics) gave positive results in 57% of cases, the retrograde stenting of the ureters was carried out in 41% of cases and the percutaneous or open nephrostomy (performed in 3 and 1 patients, correspondingly) in 2% of cases. During the management of the patients with a complicated hydronephrosis of pregnancy, the interdiscipline training of urologists and obstetricians-gynecologists as well as their close collaboration are of decisive importance. PMID- 12556632 TI - [Treatment of patients with urinary tract obstruction and significant renal impairment]. AB - Prognosis of renal function changes due to obstructive uropathy and prediction of remaining renal function after release of obstruction, have great impact on treatment we choose. There were analyzed various aspects of treatment and course of disease in 26 cases of obstructive uropathy with significant renal function impairment. It was found, that recovery of renal function after adequate kidney drainage was slower in cases of infravesical obstruction and vesicoureteral reflux in comparison with cases of ureteral obstruction. The renal function recovers more rapidly in cases with thicker parenchyma. Active surgical intervention and creation of adequate urine outflow from the obstructed kidney is method of choice at the beginning of treatment, even in cases with prolonged anamnesis of obstruction, significant renal failure and septic complications. PMID- 12556633 TI - [Chronic prostatitis with chronic pelvic pain syndrome]. AB - Almost 10% of the adult male population suffer from prostatitis. The International Prostatitis Collaborative Network has devised and validated a clinically useful classification of prostatitis that urologists and primary care clinicians will find helpful. According to this schema, chronic bacterial prostatitis is clearly an infectious disease, and patients with chronic prostatitis associated with chronic pelvic pain syndrome can have either inflammatory or noninflammatory disease. Chronic bacterial prostatitis is uncommon, chronic nonbacterial prostatitis (CPPS) is extremely common. Antibiotic therapy is indicated in management of chronic bacterial prostatitis and inflammatory chronic pelvic pain syndrome. Fluoroquinolones are safe and effective in managing chronic bacterial prostatitis. Based on literature, noninflammatory chronic pelvic pain syndrome can be treated using adrenergic blockade, analgesic, tricyclic antidepressants, benzodiazepie, physical therapy. PMID- 12556634 TI - [Treatment of 101 patients with traumatic urethral strictures by the method of V. Rusakov]. AB - Urethral strictures after the pelvic bone and perineum injury are a common cause of disability in such patients. Outcomes of several methods of surgical treatment are unsatisfactory in unacceptably high percentage of the cases. This disease seems thus to be one of the most threatening problems in urological practice. Since 1978 to 2000, 101 patients with urethral traumatic strictures were treated by the method of anastomotic urethroplasty proposed by V. Rusakov, at the Urology Department of Vilnius University Hospital. Overviewed are the classification of urethral strictures by their etiology, clinical course, degree of narrowing of the urethra, localization, length and multiplicity as well as the complications. Pre-operation work-up, instruments, operation technique and post-operative care are discussed, too. PMID- 12556635 TI - [Treatment of bladder neck obstruction (sclerosis): personal experience and literature review]. AB - The goal of this article is a description of bladder neck obstruction (sclerosis) treatment results at the Department of Urology at Kaunas University of Medicine Hospital and the review of current literature on this subject. Sixty seven patients treated for bladder neck obstruction (sclerosis) in the period of 1995 2000 years were analyzed retrospectively. On admission to the hospital, 31 patient had undergone open prostatectomy for benign prostatic hyperplasy, 14 had undergone thransurethral prostate resection and 22 had no previous prostatic surgery. The usual diagnostic procedure for bladder neck obstruction (sclerosis) in Kaunas University of Medicine Hospital is retrograde uretherocystography. According to current literature the videourodynamics is the most effective diagnostic procedure, however, still not available at Kaunas University of Medicine Hospital. The transurethral bladder neck incision or/and resection is effectively used for bladder neck obstruction (sclerosis) treatment at Kaunas University of Medicine Hospital. The near postoperative results are good for 73.4% of patients. This method is the most widely used for bladder neck obstruction (sclerosis) treatment around the world. PMID- 12556636 TI - [Piezoelectric extracorporeal lithotripsy "EDAP LT 01" for the treatment of ureteral stones]. AB - The results of 517 cases of urethral stones were analyzed concerning stone location, urethral manipulation and treatment position. The ureter was divided into six segments: ureteropelvic junction (PUJ), proximal ureter (PS1 and PS2), midureter (VS), and distal ureter (DS1 and DS2). The overall fragmentation rate was 78%. Extracorporeal piezoelectric lithotripsy combined with endourological manipulation is highly efficient in the management of ureteropelvic junction and distal ureter DS1. The advantages of the "EDAP LT" 01 are the mobility of the wave unit and rare need for general anesthesia. PMID- 12556637 TI - [Effectivity of phytotherapeutic agent prostavit for the treatment of benign prostate hyperplasia]. AB - Benign prostate hyperplasion's treatment with phytotherapic agent Prostavit and its results are described. Twenty patients who took Prostavit for 8 weeks were managed and treated. No patients were treated with placebo. Subjective and objective improvement was noticed with Prostavit, starting 6th week. Worsening of symptoms or allergical reaction was not noticed in all patients during this period. PMID- 12556638 TI - [Penis deviation correction using Schroder's method and patients' postoperative quality of life]. AB - Most frequent methods of penis deviation operative treatment are penis corporoplastics after Schroder and Nesbit. The penis corporoplastics can be followed by early complications (bleeding from the penis, recrudescent deviation due to suture cut-out, reduced or extinct sensitivity of the penis head as a result of n. dorsalis penis injury, etc.) or tardy ones (scarring phymosis, recrudescent deviation due to the scar, etc.). The objective of the study is to assess the quality of the patients' life after the operation (penis corporoplastics after Schroder) and any possible post-operative complications. The medical documentation of twelve patients, who were operated in the aforesaid manner at Kaunas 2(nd) Clinical Hospital and Kaunas Clinics of General Medical Practice in 1994-2001, was checked. Before the operation all the patients had penis deviation diagnosed (two ventral, two lateral and five mixed) resulting from congenital penis curvature. The indication for the operation of all the patients was aggravated intercourse. None of the patients showed early post operative complications. When the patients were checked 6 months and 1 year later there were no tardy post-operative complications, either. All the patients were satisfied with the quality of life after the operation - there was no deviation, the erection and intercourse were normal. Penis corporoplastics after Schroder is a proper method to be used for correction of inherent penis deviations. PMID- 12556639 TI - [Incidence of varicocele measured by echoscopy]. AB - Varicocele should be diagnosed and treatment should be started in boys under 10 14, because at an older age it will interfere with the development of the testicle. The objective of the study is to establish the rate of varicocele. The study material and methods. One hundret of patients aged from 20 to 87 (SD=17) have been studied: the thickness of veins of plexus pampiniformis was measured by echoscopy, funiculus spermaticus was palpated. RESULTS: Varicocele was diagnosed in 24 (24%) of patients. CONCLUSION: The incidence rate of this disease is high. PMID- 12556640 TI - [Laparoscopic colposuspension - effective, minimally invasive method of treatment of female urinary incontinence]. AB - THE AIM OF WORK: To describe laparoscopic method of treatment of female stress urinary incontinence, indications and contraindications for this operation, to show the first results of this operation in Lithuania, to find out the advantages of this method in comparison with open classical operations. MATERIALS AND METHODS: Ten patients were operated on laparoscopically in the period 1999-2001 due to stress urinary incontinence. The original Burch operation with sutures was performed for two patients, in 8 cases the colposuspension was performed using ligaments (prolene mesh). Transperitoneal colposuspension was performed in 9 cases, extraperitoneal - in one case. Laparoscopic cholecystectomy was performed simultaneously for 3 patients. The average time of hospitalization was 7 days, postoperative stay - 4 days. Catheter in urine bladder was removed after three days, in two cases the remaining urine was observed. RESULTS: Postoperatively in 9 cases the cystocele has disappeared, in one case the recurrence of cystocele was observed. Three months after laparoscopic colposuspension, results as very good were evaluated in 8 cases, good - in one case, satisfactory - in one case (urine continence, imperative urination with minimal urine flow). CONCLUSIONS: Laparoscopic colposuspension - effective method of treatment of stress urinary incontinence, having more advantages in comparison with open operations: shorter time of hospitalization, good postoperative results. The operation is safe, esthetic, with no postoperative hernias. It is an interesting alternative for open operations. PMID- 12556641 TI - [Percutaneous nephrostomy at the Second Kaunas Clinical Hospital]. AB - From 1996 to 2000 in urology department of 2(nd) Kaunas Clinical Hospital, 343 percutaneous interventions were performed, 250 of which was percutaneous nephrostomy. Urinary flowing was disturbed to 171 patients with urinary obstruction and 79 with malignant tumor. Death related to operation was not noticed in all patients. One of the patient had undergone surgery. PCN was successfully performed in 97.5% of all the cases. PMID- 12556642 TI - [Treatment of superficial transitional cell bladder carcinoma. Long-term results of trial comparing transurethral resection alone and adjuvant chemotherapy with Doxorubicin]. AB - OBJECTIVE: We compared the efficacy of transurethral resection alone or transurethral resection followed by bladder instillations of Doxorubicin for 1 year in patients with superficial bladder carcinoma, and followed them long term for the incidence of recurrence and progression to muscle invasion. MATERIALS AND METHODS: Between December 1998 and December 2000 a total of 69 patients with superficial transitional cell carcinoma of bladder participated in this prospective study. Final analysis of treatment results included 64 patients. Doxorubicin was administered to 25 patients, 39 patients were treated only by TUR. Patients were followed by control cystoscopy. RESULTS: The mean follow-up was 22.95 months; SD 7.79. Mean time to first recurrence in Doxorubicin group was 14.14 months; SD 7.84, in TUR alone group - 7.61 months; SD 4.4; p>0.05. Disease free survival was significantly prolonged in Doxorubicin group; p<0.05. There are no significant difference to comparison recurrence rate and progression rate between two groups. CONCLUSIONS: In regard to time of first recurrence and disease free survival this study indicates that adjuvant chemotherapy with Doxorubicin is superior to transurethral resection alone. However, progression in stage or recurrence rate was not influenced by the treatment regimen. PMID- 12556643 TI - [Antigraft response in advanced age cadaveric renal recipients]. AB - We aimed to compare graft loss due to acute rejection (AR) and chronic allograft failure (CAF) in first cadaveric kidney recipients, subsequently grafted during 1992-2000. All recipients (n=225) were divided into 3 age groups: Gr 1 (n=39) - 13-25 yrs, Gr 2 (n=169) - 26-55 yrs and Gr 3 (n=47) 56-71 yrs. There were no differences between the groups in terms of sex (m/f ratio 1.2-1.4), ischemia time and HLA matching. Incidence of high presensitization (PRA> or =50%) were observed in 15%, 5% and 9%, correspondingly (statistically significant difference was found only between Gr 1 and Gr 2). Immunosuppression consisted of CyA, AZA or MMF and steroids. Antigraft response was evaluated by graft loss due to acute rejection or chronic allograft failure. Actuarial graft survival was estimated with the Kaplan-Meier method with p values given for log-rank test. Graft loss was censored for death of recipient with functioning transplant and other causes of loss not related to rejection. Graft survival was 77%, 86% and 100% at 1yr in groups 1, 2, 3; 74%, 78% and 88% at 3 yrs; 67%, 73% and 88% at 5 yrs; 54%, 65% and 88% at 7 yrs, correspondingly (for Gr 1-3 - p<0.005, for Gr 2-3 p<0.025). In conclusion, antigraft response in elderly cadaveric kidney recipients is weaker - the incidence of transplant rejection is smaller than in young and grown-up recipients. PMID- 12556644 TI - [Transurethral microwave thermotherapy for benign prostatic hyperplasia: preliminary clinical results]. AB - We evaluated the efficacy of high energy transurethral microwave thermotherapy for treating benign prosthetic hyperplasia. During two years 42 patients with benign prostatic hyperplasia were treated with high energy transurethral microwave thermotherapy. It makes 18% of all the patients been treated. Initial evaluation consisted of a history, clinical examination, transrectal prostate ultrasonography, biopsy, PSA and urodynamic investigation with a pressure flow study. At baseline median patient age was 73 years and median prostate size was 68 g. Median 85 W were administered and treatment time 49 min. After the treatment we subjectively noticed that the patients were feeling better. The complaints according IPSS decreased to 42%, the prostate weight - 22%, removed cistostoms 75% patient, after 6 months been operated 7% of the patient. We noticed the following complications: 3 patients macrohematuria, 1 - orchoepidydymitis, 1 - acute retention of the urination. TUMT has some advantages comparing with other. It is the minimal invasion, the procedure is less painful, less hemorrhage, less complicated numbers. This method is not often used, because there are some contraindication. For patients, who need operation, the age is longer, because of the advance of good medicinal therapy. Transurethral hyperthermia represents a safe and promising method to treat benign prostatic hyperplasia, particularly for patient who candidates for conventional surgical approaches because of medical or personal reasons. PMID- 12556645 TI - [Results of the kidney autotransplantation]. AB - We report our experience with kidney autotransplantation and evaluate the benefit of this operative method in selected patients. Between 1991 and 2001, 8 patients underwent kidney autotransplantation in our department, 2 involving iatrogenic ureter lesion, 3 - gross hematuria due to kidney vascular abnormalities, 2 - renal arteries stenoses, 1 with long ureteric stenose. In short-term follow-up period 7 patients had normal kidney function (serum creatinine level was 99.5 micromol/l), 1 patient had impaired kidney function (serum creatinine content was 170 micromol/l), serum creatinine level improved in 4 patients and were stable in another 4 patients, 3 patients experienced relief gross hematuria. In long-term follow-up period 5 patients had normal kidney function and no experienced hematuria episodes. We concluded that kidney autotransplantation is feasible and effective method for the surgical treatment with good results for selected patients with ureteric lesions, renovascular disorders, it also appears to be effective treatment for patients with gross hematuria. Kidney autotransplantation allows to recover or stabilise impaired kidney function due to renovascular or ureteric disorders and to avoid nephrectomy due to gross hematuria, due to vascular abnormalities. PMID- 12556646 TI - Results of the ALLHAT trial: is the debate about initial antihypertensive drug therapy over? PMID- 12556647 TI - The ALLHAT report: a case of information and misinformation. PMID- 12556648 TI - Opportunity for JNC VII. PMID- 12556649 TI - The effects of amlodipine compared to losartan in patients with mild to moderately severe hypertension. AB - The calcium channel blocker amlodipine and angiotensin II receptor blocker losartan, with or without hydrochlorothiazide (HCTZ), were compared for the treatment of mild to moderate hypertension in a multicenter, double-blind, parallel-group clinical trial. Following a 2-week placebo run-in, 440 adults (45 80 years old) were randomized to receive either amlodipine 5 mg once daily or losartan 50 mg once daily. Patients who failed to meet the sitting diastolic blood pressure (BP) reduction goal of 350 grams. Left ventricular systolic dysfunction was seen in 9% of the men with uncontrolled hypertension, and none of the men with controlled hypertension (p=0.02). Renal dysfunction was found in 12% of the subjects, and microalbuminuria or gross proteinuria in 34%. The authors conclude that there is a high prevalence of cardiac and renal abnormalities in inner-city African American men with hypertension, especially in men on antihypertensive therapy with uncontrolled hypertension. It is imperative that cost-effective medications and culturally acceptable health care delivery programs be developed, tested, and integrated into health systems, with strategies specifically relevant to this high-risk population, to decrease the largely preventable morbidity and mortality associated with hypertension. PMID- 12556651 TI - How pharmacists can assist physicians with controlling blood pressure. AB - The goals for hypertension in the publication Healthy People 2010 require a much more intensive approach in order to achieve desired blood pressure control rates. The pharmacist is uniquely positioned in the health care system to assist with improving blood pressure control by utilizing strategies to solve medication related problems. Studies within integrated health systems have demonstrated that when pharmacists are included as members of health care teams, control rates for hypertension increase. In addition, drug interactions, nonadherence, and costs can be reduced. The authors suggest that incorporating a pharmacist or some services typically provided by traditional pharmacists into physician practices can improve blood pressure control. This review summarizes studies involving pharmacist participation in hypertension management and provides recommendations for obtaining pharmacist involvement. PMID- 12556652 TI - Hypertension in women: the Women Take Heart project. AB - Hypertension is an important, modifiable risk for cardiovascular disease. The Women Take Heart study, a prospective, community-based cohort study of risk factors for heart disease, provides an opportunity to examine prevalence, awareness, and control of hypertension specifically in women. In 1992, 5932 women, age 35 and older (mean age, 52.9; 86% white, 9% African American, 5% other) and free of active heart disease symptoms for 3 months, were recruited through Chicago area public announcements, and their baseline examination data analyzed. Overall, 47.6% were hypertensive (systolic blood pressure >or=140 mm Hg or diastolic blood pressure >or=90 mm Hg, or self-report). Only 17.3% reported being hypertensive; in 63.2% of all hypertensive women, the hypertension was undetected or unacknowledged. Blood pressure was controlled to <140/90 mm Hg in 24.1% of self-reported hypertensives. Results from this study and national surveys indicate that hypertension detection and control remain major public health challenges in preventing cardiovascular disease in older women. PMID- 12556653 TI - The relationship between systolic blood pressure and cardiovascular risk--results of the Brisighella Heart Study. AB - We examined the relationship of systolic (SBP) and diastolic (DBP) blood pressure, and pulse pressure to coronary heart disease and cerebrovascular disease risk in a prospective population-based European cohort. The Brisighella Heart Study included 2939 men and women between the ages of 14-84 without prior coronary heart disease or cerebrovascular disease and not taking antihypertensive therapy at baseline. Cox regression was used to obtain hazard ratios (HRs) for coronary heart disease and cerebrovascular disease as a function of baseline blood pressure parameters over a 23-year follow-up. Higher combined coronary heart disease and cerebrovascular disease risk was evident in comparison to the referent of <120 mm Hg, with a 44% increased risk at SBP 120-139 mm Hg (HR, 1.44; 95% confidence interval [CI], 1.00-2.09; p=0.052), 76% increased risk at SBP 140 159 mm Hg (HR, 1.76; 95% CI, 1.16-2.69; p=0.009), and 109% increased risk at SBP >or=160 mm Hg (HR, 2.09; 95% CI, 1.31-3.35; p=0.0021). Trends of increasing risk with increasing levels of blood pressure were significant for SBP and pulse pressure, (p<0.0001) but not for DBP (p=0.058). In this European cohort, SBP was a stronger predictor of coronary heart disease and cerebrovascular disease events than DBP, and an increase in risk was already evident with high-normal SBP (120 139 mm Hg). The prognostic significance of pulse pressure was also demonstrated. The importance of SBP as seen in the Framingham Heart Study may be generalized to a European population with differences in diet and other risk factors. PMID- 12556654 TI - The effect of nonsteroidal anti-inflammatory drugs on blood pressure in patients treated with different antihypertensive drugs. AB - Hypertension and arthritis are both common diseases in the older age group and require pharmacologic treatment. Nonsteroidal anti-inflammatory drugs (NSAIDs) alter renal function if given in high enough doses, reducing renal blood flow and the glomerular filtration rate and causing sodium retention. In salt sensitive subjects, this retention of sodium will cause blood pressure to rise. Salt sensitivity is more common in elderly patients, in diabetics, and in people with renal failure. When most antihypertensive drugs are used, people become salt sensitive, as shown by the additive effect of salt restriction or diuretics on blood pressure response. The responses to dihydropyridine and possibly other calcium channel blocking drugs are not affected to any major extent by sodium intake or by diuretics. Studies are described which indicate that indomethacin elevates blood pressure in elderly people treated with enalapril, but not in people whose blood pressure is controlled with amlodipine or felodipine. It is unclear whether the various NSAIDs have different effects on blood pressure. It is proposed that if the same analgesic effect is achieved with the same amount of cyclooxygenase inhibition, the response will be similar. Aspirin, used in a prophylactic dose, does not inhibit to this extent and does not elevate blood pressure. If elderly people require NSAIDs, it would appear that dihydropyridine calcium channel blocking drugs are more effective at lowering and maintaining blood pressure control and should be one of the drugs used. If patients are on other antihypertensive agents, it is important to monitor blood pressure when a NSAID is added to therapy. PMID- 12556655 TI - Evaluation of angiotensin II receptor blockers for 24-hour blood pressure control: meta-analysis of a clinical database. AB - Angiotensin II receptor blockers are well tolerated and improve compliance in hypertensive patients. The need for 24-hour blood pressure control has focused attention on whether all agents in this class maintain smooth antihypertensive effects over a 24-hour period. Insight into this issue emerged from a meta analysis of five large, multicenter trials in which ambulatory blood pressure monitoring was used to compare the antihypertensive effects of three angiotensin II receptor blockers: telmisartan, losartan, and valsartan. These trials used either a double-blind, placebo-controlled or a prospective, randomized, open label, blinded-end point design. Initial analysis established the validity of combining ambulatory blood pressure monitoring data from the double-blind, placebo-controlled and prospective, randomized, open-label, blinded-end point designs. Subsequent analyses revealed that telmisartan 80 mg was significantly more effective than losartan 50 mg and valsartan 80 mg for reducing 24-hour mean blood pressure. Furthermore, telmisartan 80 mg was comparable to amlodipine 5 mg for controlling the early morning surge in blood pressure. PMID- 12556656 TI - Meta-analysis, meta-regression, and meta-physics. PMID- 12556657 TI - Benefits of antihypertensive pharmacologic therapy and blood pressure reduction in outcome trials. AB - In a quantitative overview of published trials, we investigated whether pharmacologic properties of antihypertensive drugs, as opposed to reduction in blood pressure, explain cardiovascular outcomes in hypertensive or high-risk patients. We used meta-regression to investigate the association between the odds ratios of outcome (experimental vs. reference treatment) and the corresponding blood pressure differences between study groups. Thus, we correlated odds ratios with between-group differences in systolic pressure. We then compared odds ratios of benefit observed in recent trials with those predicted by meta-regression on the basis of the differences in systolic pressure between randomized groups. Among nine actively-controlled trials in hypertension, significant differences in systolic pressure (follow-up minus baseline) between randomized groups (experimental minus reference) were observed in the ALLHAT, CAPPP, MIDAS, and NORDIL trials. Furthermore, the differences in achieved systolic and/or diastolic pressure between study groups were also significant in the hypertension trials and studies in high-risk patients, which involved untreated control patients. The differences between the observed odds ratios and those predicted by meta regression did not reach statistical significance except for NORDIL and the single-drug therapy subgroup of the PROGRESS trial. In NORDIL, the risk of stroke was lower on diltiazem than on the older drug classes despite a 3.1 mm Hg higher systolic pressure on the calcium channel blocker. In PROGRESS, perindopril alone reduced blood pressure by 5/3 mm Hg, but did not affect the incidence of all cardiovascular events or the recurrence of stroke. In conclusion, the finding that in the reviewed trials blood pressure reduction largely accounted for outcome emphasizes the desirability of tight blood pressure control. The hypothesis that blood pressure-lowering medications might influence cardiovascular prognosis over and beyond their antihypertensive effect remains to a large extent unproved. PMID- 12556658 TI - Spontaneous renal hematoma (Wunderlich syndrome) associated with severe hypertension. PMID- 12556663 TI - Drug-induced hypertension. PMID- 12556664 TI - Men are from Mars, women are from Venus: stress, pets, and oxytocin. PMID- 12556665 TI - The forgetful hypertensive patient. PMID- 12556666 TI - Introduction: management of hypertension and cardiovascular risk factors in African Americans. PMID- 12556667 TI - Epidemiology of hypertension and cardiovascular disease in African Americans. AB - Hypertension is a major cause of cardiovascular-renal morbidity and mortality and all-cause mortality. It is a highly significant problem for African Americans; about 30% of all deaths in this population are attributable to hypertension. Compared with whites, hypertension in African Americans is more prevalent, occurs earlier in life, is more severe, and is more often associated with target organ injury such as left ventricular hypertrophy and other cardiovascular complications. Only 25% of all African Americans with hypertension and fewer than 50% of those receiving drug treatment attain a blood pressure <140/90 mm Hg. These control rates are somewhat less than in white Americans. Enhanced awareness and understanding of the epidemiologic patterns of hypertension, other cardiovascular risk factors, risk-factor control rates, and factors influencing these control rates should lead to better approaches to risk-factor control. This most likely would result in a reduction of cardiovascular disease complications. PMID- 12556668 TI - Blockade of the renin-angiotensin system in African Americans with hypertension and cardiovascular disease. AB - There is overwhelming evidence that the renin-angiotensin system plays a significant role in the pathophysiology of hypertension and target organ damage. Agents that regulate the renin-angiotensin system, such as angiotensin-converting enzyme (ACE) inhibitors, angiotensin II receptor blockers, and aldosterone antagonists, are not only effective antihypertensive agents but can prevent target organ damage. Although diuretics remain the agents of first choice for the treatment of hypertension in African Americans, ACE inhibitors have a clear role in the management of these patients. ACE inhibitors (usually when used with a diuretic) have been shown to reduce morbidity and mortality in a wide range of patient groups. ACE inhibitors are infrequently used in African Americans because of a belief that these agents are ineffective in this racial group; however, when adequate dosing and appropriate combinations are used, ACE inhibitor therapy provides effective blood pressure control. In particular, the addition of diuretics to ACE inhibitor therapy ameliorates the racial differences in efficacy seen when ACE inhibitors are administered as monotherapy. Although further confirmation in additional clinical trials is required, increased use of these agents in African Americans will likely result in a reduction in target organ damage. PMID- 12556669 TI - Optimal treatment of hypertension and cardiovascular risk reduction in African Americans: treatment approaches for outpatients. AB - African Americans have a higher prevalence and greater severity of hypertension compared with whites and therefore have a higher prevalence of many disease related complications, such as coronary heart disease, stroke, and end-stage renal disease. Minorities have been, until recently, underrepresented in large clinical trials, leading to a lack of outcome data for these patient groups. However, accumulating data confirm the benefit of aggressive blood pressure lowering therapy in this population. These studies also show that most patients require combination therapy to achieve adequate blood pressure reduction, particularly high-risk patient groups that have lower target blood pressure goals. All of the available antihypertensive agents are effective in African Americans. Recent studies suggest that regimens containing a thiazide-type diuretic are unsurpassed in blood pressure lowering and prevention of major clinical complications, and they cost less. Thus, while other agents may be required for selected clinical indications or for blood pressure control, diuretics should be drugs of first choice or included in most antihypertensive regimens especially in African American hypertensives. PMID- 12556670 TI - Treatment of high-risk African American patients: left ventricular dysfunction, heart failure, renal disease, and postmyocardial infarction. AB - African Americans experience more mortality and morbidity from hypertension related complications than other racial groups. Although angiotensin-converting enzyme (ACE) inhibitors have clearly been shown to reduce mortality and morbidity in hypertensive white patients with heart failure, renal dysfunction, stroke, and acute myocardial infarction, African American patients have been underrepresented in these trials. The lack of direct evidence of the benefit of ACE inhibitors in these individuals and the suggestion that ACE inhibitors are less efficacious in this group has resulted in a reluctance to use ACE inhibitors in African Americans. However, retrospective analyses in black patients with heart failure and a recent randomized clinical trial in African Americans with renal dysfunction suggest that a regimen based on ACE inhibitors is efficacious in this racial group. Although diuretics remain first-line therapy, data now suggest that ACE inhibitors provide additional benefit and should be considered for use in patients with high-risk complications regardless of race. PMID- 12556671 TI - Therapeutic lifestyle changes for hypertension and cardiovascular risk reduction. AB - Elevated blood pressure is the most common chronic illness in the United States, affecting more than 50 million people. Hypertension is an even greater problem in the African American community. Traditionally, management of hypertension and cardiovascular risk reduction has focused on drug therapy; however, several studies have shown the benefits of therapeutic lifestyle changes for blood pressure lowering and cardiovascular risk reduction. Therapeutic lifestyle changes to reduce blood pressure have enormous potential as a means for preventing and controlling hypertension and thereby reducing the risk of coronary heart disease. Although the reductions in blood pressure are relatively modest with these approaches, they could potentially have a beneficial impact on overall cardiovascular morbidity and mortality when applied to the whole population. Because of their high prevalence of certain cardiovascular risk factors (e.g., obesity, diabetes mellitus) and greater salt sensitivity, therapeutic lifestyle changes have particular relevance for African Americans. PMID- 12556672 TI - Potential role of type 5 phosphodiesterase inhibition in the treatment of congestive heart failure. AB - Endothelial dysfunction is associated with impairment of aerobic capacity in patients with heart failure and may play a role in the progression of disease. Impaired endothelium-dependent vasodilation in patients with heart failure can be attributed to decreased bioavailability of nitric oxide and attenuated responses to nitric oxide in vascular smooth muscle. Impaired vasodilation in response to nitric oxide derived from vascular endothelium or organic nitrates in vascular smooth muscle may be related in part to increased degradation of the second messenger cyclic guanosine monophosphate by type 5 phosphodiesterase. Sildenafil, a specific type 5 phosphodiesterase inhibitor currently approved for the treatment of erectile dysfunction, has been shown to acutely enhance endothelium dependent vasodilation in patients with heart failure. Further studies are warranted to characterize the safety and efficacy of type 5 phosphodiesterase inhibition in the treatment of chronic heart failure. PMID- 12556673 TI - Nontransplant surgical options for congestive heart failure. AB - A wide array of surgical options are currently available for the treatment of congestive heart failure ranging from traditional coronary artery bypass grafting to total artificial heart implantation. The indications for each procedure depend on the severity of disease and the individual patients desires. Some surgical options are indicated for patients with moderate disease and prevent worsening heart failure, whereas other procedures are limited to patients who will only survive with high-risk surgery. Ongoing technologic advances are increasing the number of patients that benefit from the reparative surgical treatment of congestive heart failure. PMID- 12556674 TI - The role of norepinephrine in exercise impairment in congestive heart failure. AB - Congestive heart failure is a disorder that includes a multitude of neurohormonal responses that become maladaptive over time. Chronic sympathetic stimulation adversely affects the well-being and survival of heart failure patients and contributes to the exercise intolerance frequently seen in these patients. Norepinephrine levels have been correlated with poorer survival in heart failure patients. Administration of norepinephrine has been shown to impair exercise responses in those with congestive heart failure, and the recent effort to incorporate beta blocker therapy into the standard management of heart failure patients addresses this abnormal neurohormonal process. Studies with central acting sympatholytics have shown mixed results. The use of drugs such as clonidine has been suggested as potentially useful therapy in the long-term management of patients with heart failure, but definitive conclusions await further study. Regular exercise has been shown to reduce resting norepinephrine levels in heart failure subjects. This may serve as an additional rationale to recommend chronic exercise for these patients. PMID- 12556675 TI - The role of angiotensin II receptor blockers in the treatment of heart failure patients. AB - Evidence from large, randomized, controlled clinical trials supports the use of angiotensin-converting enzyme (ACE) inhibitors, beta blockers, and spironolactone to reduce mortality and morbidity. Despite these effective therapies, event rates related to heart failure remain high. Although ACE inhibitors reduce angiotensin II production, they do not fully suppress the increased angiotensin II production in heart failure. Angiotensin II receptor blockers (ARBs) directly block the effect of angiotensin II, derived from any source, at the receptor level and have the potential to be as effective or even more effective than ACE inhibitors. The results of a number of clinical studies have demonstrated ARBs are effective and well tolerated. However, no studies have demonstrated a convincing decrease in mortality with ARB use, although a decrease has been observed for heart failure hospitalization. The results from further studies are awaited to clarify the role of ARBs in the treatment of heart failure. PMID- 12556676 TI - Cardiogenic shock: a summary of the randomized SHOCK trial. AB - Cardiogenic shock is the most common cause of death for patients hospitalized with acute myocardial infarction. The Should We Emergently Revascularize Occluded Coronaries for Cardiogenic Shock (SHOCK) trial randomly assigned 302 patients with predominant left ventricular failure following an acute myocardial infarction to a strategy of emergency revascularization or initial medical stabilization. Emergency revascularization by either coronary artery bypass grafting or angioplasty was required within 6 hours of randomization. Patients assigned to initial medical stabilization could undergo delayed revascularization at a minimum of 54 hours post-randomization. The primary end point of the study was 30-day all-cause mortality. Overall survival at 30 days did not differ significantly between the emergency revascularization and initial medical stabilization groups (53% vs. 44%; p=0.109). However, at the 6- and 12-month follow-up, there was a significant survival benefit with early revascularization (50% vs. 37%; p=0.027 and 47% vs. 34%; p=0.025, respectively). The benefit appeared to be greatest for those less than 75 years of age, with 20 lives saved at 6 months per 100 patients treated. According to the results of the SHOCK trial, the American College of Cardiology/American Heart Association guidelines for myocardial infarction now recommend emergency revascularization for patients younger than 75 years with cardiogenic shock. PMID- 12556677 TI - Hyperthyroidism: a "curable" cause of congestive heart failure--three case reports and a review of the literature. AB - With the increasing incidence of coronary artery disease and the aging population, the prevalence of congestive heart failure (CHF) is increasing. In the majority of these cases the etiology is underlying coronary artery disease. Other less common causes of CHF include valvular heart disease, hypertension, alcoholic cardiomyopathy, and dilated cardiomyopathy. In addition, there are rare causes, one of which is hyperthyroidism. Hyperthyroidism can affect the cardiovascular system in a variety of ways. The cardiovascular manifestations range from sinus tachycardia to atrial fibrillation and from a high cardiac output state to CHF due to systolic left ventricular dysfunction. If the underlying hyperthyroidism is recognized and treated early the CHF in such cases can be cured. The authors present three cases of CHF due to systolic left ventricular dysfunction secondary to hyperthyroidism, which showed considerable improvement in the left ventricular function once the hyperthyroidism was treated. PMID- 12556681 TI - Social health inequalities during the course of chronic HIV disease in the era of highly active antiretroviral therapy. PMID- 12556678 TI - Difficult cases in heart failure: chemotherapy-related cardiomyopathy--all that glitters is not gold. AB - As long-term morbidity with chemotherapeutic exposure becomes a clinical reality, the development of cardiomyopathy with heart failure is rapidly growing. The type of chemotherapy agent, dose, rate of administration, use of combination chemotherapy, and concomitant radiation therapy are major factors that define the propensity to cardiotoxicity. Chemotherapy-related cardiomyopathy is multifactorial in origin and requires a diligent etiologic survey to arrive at the appropriate diagnosis. A contemporary discussion of chemotherapy-related cardiomyopathy with particular emphasis on the pathogenesis and ameliorative targets of this unique clinical disorder are described in this report. PMID- 12556682 TI - CD4-dependent and CD4-independent HIV-2: consequences for neutralization. AB - BACKGROUND: HIV-2 is less pathogenic than HIV-1. In contrast to HIV-1, many isolates of HIV-2, including primary isolates, can infect cells independently of CD4. OBJECTIVE: To compare the sensitivity of CD4-dependent and CD4-independent isolates of HIV-2 to antibody-mediated neutralization. METHODS: The neutralization sensitivity of CD4-dependent and CD4-independent molecular clones of HIV-2 to a panel of HIV-2-positive serum samples was tested. Monoclonal antibodies to various epitopes across the viral envelope were used to determine whether a specific epitope conferred neutralization sensitivity. Neutralization sensitivity of primary isolates of HIV-2 able to infect in the absence of cellular CD4 was also investigated. Antibody binding to sensitive and resistant envelopes was analysed using enzyme-linked immunosorbent assay and flow cytometry. RESULTS: CD4-independent ROD B was highly sensitive to neutralization by HIV-2-positive sera compared with the CD4-dependent isolate ROD A. Induction of ROD A to infect CD4-negative cells by soluble CD4 rendered it equally sensitive to antibody neutralization. Similarly, primary X4, R5 or dual-tropic isolates of HIV-2 were significantly more susceptible to neutralization when utilizing a CD4-independent route of infection. Neutralization sensitivity was not epitope specific but several conformation-dependent antibodies accentuated this phenotype. Antibody binding to monomeric or oligomeric envelope did not correlate with neutralization sensitivity. CONCLUSIONS: HIV-2 isolates utilizing a CD4-independent route of infection are more sensitive to antibody-mediated neutralization. Cellular CD4 may protect HIV-2 from neutralization. This sensitivity to neutralization may, in part, explain the lower virus load and slower progression to disease in HIV-2-infected individuals. PMID- 12556683 TI - Post-exposure prophylaxis with human monoclonal antibodies prevented SHIV89.6P infection or disease in neonatal macaques. AB - BACKGROUND: The majority of infants infected through maternal transmission acquire the virus during birth or postpartum through breastfeeding: mucosal exposure is considered to be a major route of infection. OBJECTIVES: To develop passive immunization with human neutralizing monoclonal antibodies (mAbs) against mother-to-child transmission of HIV during delivery and through breastfeeding. DESIGN: An oral challenge model in newborn rhesus macaques mimicked peri- and postpartum virus transmission. METHODS: Neonatal rhesus macaques were challenged orally with the highly pathogenic, chimeric simian-human immunodeficiency virus SHIV89.6P and given post-exposure prophylaxis with a quadruple combination of neutralizing human mAbs, IgG1b12, 2G12, 2F5, and 4E10, directed against conserved epitopes of HIV envelope glycoproteins. Control animals were virus challenged but left untreated. All infants were followed prospectively for signs of viremia and immunodeficiency. RESULTS: Two out of four macaque infants treated with neutralizing mAbs showed no evidence of infection; the other two maintained normal CD4 T cell counts. In contrast, all control animals became highly viremic and had profound CD4 T cell losses; three out of four died from AIDS within 1.5-6 weeks of the challenge. CONCLUSIONS: Passive immunization with this quadruple neutralizing mAbs combination may represent a promising approach to prevent peri- and postnatal HIV transmission. Furthermore, the epitopes recognized by the four neutralizing mAbs are key determinants to achieve complete protection and represent important targets against which to develop active, antibody-response based AIDS vaccines. PMID- 12556684 TI - Human papillomavirus infection and abnormal cytology of the anus in HIV-infected and uninfected adolescents. AB - OBJECTIVE: To examine the prevalence of and risk for anal human papillomavirus (HPV) infection and abnormal anal cytology in sexually active adolescents. DESIGN: Prevalence data from adolescents aged 13-18 years with and without HIV infection and with a history of high-risk sexual behavior. METHODS: HPV DNA was detected using amplification techniques. Abnormal anal cytology was defined as atypical squamous cell of undetermined significance or worse. RESULTS: Prevalence of anal HPV infection was similar in HIV-infected [28/58 (48%)] and uninfected [9/25 (36%)] boys (P = 0.3). but greater in HIV-infected [59/183 (59%)] than in uninfected [11/82 (13%)] girls (P < 0.001). Perianal warts were a risk for anal HPV in both boys [odds ratio (OR), 15.5; 95% confidence interval (CI), 1.6-149] and girls (OR, 9.9; 95% CI, 1.9-51.3). In subjects without anal warts, HIV infection was significant for girls (OR, 2.3; 95% CI, 1.1-4.9) and homosexual/bisexual orientation was significant for boys (OR, 5.2; 95% CI, 1.3 20.6). Abnormal anal cytology was more common among boys [32/77 (41.6%)] than girls [38/230 (16.5%)] (P < 0.001) and in addition to anal HPV, independent risk factors were positive HIV status in boys (OR, 6.5; 95% CI, 1.5-11.9) and number of partners within the past 3 months in girls (OR, 4.2; 95% CI, 1.5-11.9). CONCLUSIONS: Strong risk factors for abnormal anal cytology were HIV infection and anal HPV in boys and anal HPV and higher number of sexual partners for girls. The results suggest that anal cytology screening should be considered in HIV infected homosexual/bisexual males. PMID- 12556685 TI - Safety and tolerability of vaginal PRO 2000 gel in sexually active HIV-uninfected and abstinent HIV-infected women. AB - OBJECTIVES: To evaluate once or twice daily vaginal exposure to 2 and 4% PRO 2000 Gel, a naphthalene sulfonate polymer microbicide, in sexually active HIV uninfected women to determine the highest tolerated frequency and concentration combination, and to assess this in sexually abstinent HIV-infected women. METHODS: Sixty three women from Providence, Philadelphia, Durban and Johannesburg were enrolled after being screened to exclude pre-existing illnesses and were instructed to use the product once or twice daily for 14 intermenstrual days. They underwent colposcopy prior to product use and after 14 days of product use, with a pelvic examination at day 7. RESULTS: The product was well tolerated, with no serious adverse events, even though 73% of the participants had at least one adverse experience: 82% of these were classified as mild, and over 90% of the findings and symptoms were localized to the genital tract. Women who used the 4% gel twice daily tended to have more adverse events than all the other groups. Three participants did not complete the study; one because of Herpes simplex virus cervicitis, the second because of epithelial disruption, and the third because she became pregnant. The remaining participants adhered to the study protocol and indicated that they would use the product if it were shown to be effective. CONCLUSIONS: PRO 2000 Gel was safe and well tolerated in sexually active HIV-uninfected and sexually abstinent HIV-infected women, enabling the product to be considered for evaluation in efficacy trials. PMID- 12556686 TI - A new quantitative HIV load assay based on plasma virion reverse transcriptase activity for the different types, groups and subtypes. AB - BACKGROUND: Plasma viral load monitoring is an integral part of the standard of care for HIV-infected patients in industrialized countries. In developing countries, viral load assay is either unaffordable or hindered by on-site maintenance and/or technical problems. OBJECTIVES: To evaluate a new and simple quantitative assay for plasma HIV reverse transcriptase (RT) activity; and to compare RT activity-based and RNA-based quantification in plasma samples from patients infected by different subtypes of HIV-1 group-M, HIV-1 group-O and HIV 2. METHODS: The RT-based viral load assay involves separation of the virion protected RT and quantification of its activity with an enzyme immunoassay. Plasma viraemia was quantified both by RT activity and by RNA copies in 322 samples from 236 HIV-1 group M-infected patients, including serial samples from 54 patients. Samples from 49 patients infected by HIV-1 group O or HIV-2 were also tested. RESULTS: RT activity and RNA copies were detected in 70% of plasma samples; respectively 25% and 1% of samples contained detectable RNA copies or RT activity alone. Measured RT activity corresponded to 48%, 96% and 100% of samples with 1.7-4.0 log(10), 4.1-4.8 log(10) and 4.9-6.7 log(10) RNA copies/ml, respectively. The values of the two assays correlated independently of the HIV subtype (P < 0.0001) and group/type (P < 0.03). Patient follow-up showed a similar pattern of viraemia with the two assays. CONCLUSION: Plasma RT activity assay is a simple, cheap and reliable alternative for HIV viral load determination. As such, it could be particularly valuable for diagnosis and treatment monitoring in developing countries. PMID- 12556687 TI - Discordant phenotypes and genotypes of cytomegalovirus (CMV) in patients with AIDS and relapsing CMV retinitis. AB - OBJECTIVE: To investigate the correlation between genotypic studies performed on blood leukocytes and phenotypic results obtained from the corresponding blood viral isolates in AIDS patients with relapsing cytomegalovirus (CMV) retinitis. METHODS: Sequential blood samples were collected from patients failing intravenous or oral ganciclovir therapy. The CMV UL97 gene was amplified directly from leukocyte DNA extracts for assessing the presence of viral mutations using restriction fragment length polymorphism analysis and direct sequencing. Positive viral cultures from the same blood samples were also analyzed for their susceptibility to ganciclovir and their UL97 genotype was determined. RESULTS: Discordant CMV genotypes between the clinical specimen and the viral culture were found in at least one blood sample from three of the four patients with relapsing CMV retinitis. Furthermore, some UL97 mutations at known resistance codons (592, 594) were associated with a drug-susceptible phenotype. In all four cases, genotypic analyses of blood samples better correlated with clinical progression than phenotypic analyses of viral cultures. CONCLUSIONS: The presence of mixed viral populations in blood samples of AIDS patients and the potential selection bias introduced by susceptibility testing may underestimate the real impact of CMV resistance in patients failing antiviral therapy. PMID- 12556688 TI - Effects of interleukin-2 therapy combined with highly active antiretroviral therapy on immune restoration in HIV-1 infection: a randomized controlled trial. AB - BACKGROUND: Intermittent interleukin-2 (IL-2) therapy leads to a sustained increase of CD4 T cells in HIV-1-infected patients. METHODS: Symptom-free HIV-1 infected patients who were naive to all antiretroviral drugs (n = 68) and/or to protease inhibitors (n = 50) and had a CD4 cell count of 200-550 x 10(6) cells/l were randomly assigned to start lamivudine/stavudine/indinavir alone (controls) or combined from week 4 with subcutaneous IL-2 (5 x 10(6) IU twice daily for 5 days: every 4 weeks for three cycles, then every 8 weeks for seven cycles). Immunological and virological results were monitored until week 74. RESULTS: CD4 T cell counts increased more in the IL-2 group than in the controls (median increases 865 and 262 x 10(6) cells/l, respectively; P < 0.0001); an 80% increase in CD4 T cells was achieving by 89% of the IL-2 group and by 47% of the controls (P < 0.0001). Decrease of plasma viral loads was similar in both groups. Compared with controls, IL-2 induced a greater increase of naive and memory CD4 T cells, lymphocyte expression of CD28 and CD25 (P < 0.0001) and natural killer cells (P < 0.001). In a logistic regression analysis, odds of being responders to recall antigens was 8.5-fold higher in IL-2 recipients (P = 0.002) than in controls. The former experienced a higher level of antibody response to tetanus vaccination at week 64 than controls (32 and 8 haemagglutinating units/ml, respectively; P = 0.01). CONCLUSIONS: The combination of antiviral drugs and IL-2 induced a greater expansion and function of CD4 T cells than antiretroviral drugs alone. PMID- 12556689 TI - Effectiveness of highly active antiretroviral therapy in Spanish cohorts of HIV seroconverters: differences by transmission category. AB - OBJECTIVE: To evaluate the population effectiveness of highly active antiretroviral therapy (HAART) in HIV progression and determine the heterogeneity of the effect of HAART in GEMES (Spanish multicenter study of seroconverters). DESIGN: Multicenter cohort study. METHODS: Data from 1091 persons with well documented HIV seroconversion dates from 1980s to January 2000 were analysed. Risk of AIDS and death in subjects with same duration of HIV infection were compared in different calendar periods; before 1992, 1992-1995 (reference), 1996 1997, 1998 and 1999 with Kaplan-Meier methods and Cox proportional hazards models, allowing for late entry, fitting calendar period as time-dependent covariate and adjusting for transmission category, age and gender. RESULTS: Statistically significant reductions in the risk of AIDS were first observed in 1998 [hazard ratio (HR), 0.59; 95% confidence interval (CI), 0.35-1.01] becoming more pronounced in 1999 (HR, 0.45; 95% CI, 0.24-0.84). Reduction in the risk of death was seen in 1997, though only reached borderline significance in 1999 (HR, 0.53; 95% CI, 0.26-1.07). Progression to AIDS and death was slower in women (HR, 0.68; 95% CI, 0.46-0.99 and HR, 0.53; 95% CI, 0.33-0.87, respectively). Compared with men who have sex with men (MSM), intravenous drug users (IDU) had lower reductions in the risk of AIDS and death. CONCLUSIONS: Reductions in incidence of AIDS and death in GEMES are seen after 1998 and 1999, respectively, compared with 1992-1995, being more pronounced in MSM compared with IDU, the commonest category in Spain. PMID- 12556690 TI - Persistence of drug-resistant HIV-1 after a structured treatment interruption and its impact on treatment response. AB - OBJECTIVE: Among treated patients with drug-resistant viremia, structured treatment interruptions often result in the re-emergence of drug-susceptible HIV 1. Theoretically, this may allow for a more durable response to salvage therapy. We therefore studied the long-term treatment outcome to antiretroviral therapy in a cohort of patients who had previously interrupted therapy, focusing on the determinants of treatment success versus failure. DESIGN: A prospective observational study of the response to antiretroviral therapy in patients resuming therapy after a treatment interruption. Virological and immunological studies were performed every month for 3 months and then every 3 months. RESULTS: Twenty-four patients underwent a structured treatment interruption and resumed therapy after a variable period of time (median 20 weeks). The median duration of treatment after the treatment interruption was 109 weeks. A transient virological response was observed in all patients who resumed a regimen containing no drug to which their pre-interruption virus was fully susceptible. Virus isolated during virological failure was genotypically and phenotypically identical to the pre interruption virus, exhibited reduced replicative capacity, and replicated at levels similar to the pre-interruption baseline. In contrast, durable viral suppression (< 200 copies/ml) was observed in patients who initiated a regimen containing only one drug to which their pre-interruption virus was fully susceptible. Despite viral suppression, the pre-interruption drug-resistant virus population remained detectable in two patients. CONCLUSION: Although drug resistant HIV-1 persists at low levels during and after the interruption of therapy, durable suppression of this virus population may be achieved with a combination regimen containing only one fully active agent. PMID- 12556691 TI - HIV-related lung cancer in the era of highly active antiretroviral therapy. AB - OBJECTIVES: To address the impact of highly active antiretroviral therapy (HAART) on the incidence and outcome of patients with HIV-related lung cancer. DESIGN AND SUBJECTS: Patients with HIV-related lung cancer were identified from a prospective HIV data base of 8400 patients diagnosed between 1986 and 2001. Patients diagnosed with HIV-related lung cancer before 1996 were in the pre-HAART cohort whereas the remainder were in the post-HAART cohort. METHODS: The incidence of HIV-related lung cancer in the pre- and post-HAART cohorts was compared with the age and sex-matched population of south east England. Clinicopathological features, treatments and outcomes were also recorded. RESULTS: The incidence of HIV-related lung cancer increased from 0.8 (95% CI 0.2 3.2)/10(5) patient-years follow-up in the pre-HAART era to 6.7 (95% CI 3.1 13.9)/10(5) patient-years follow-up in the post-HAART era. The age and sex matched incidence of lung cancer in south east England was 0.75 (95% CI 0.63 0.87)/10(5) patient-years, suggesting that HIV-related lung cancer only occurred more frequently in the post-HAART era (relative risk 8.93, 95% CI 4.92-19.98). The patient characteristics and outcomes were similar in the pre- and post-HAART eras, although the time interval between testing HIV positive and developing HIV related lung cancer was longer in post-HAART patients. CONCLUSION: In this study HIV-related lung cancer occurred more frequently in the post-HAART era, when compared with the HIV-negative population. Unfortunately, the outcome of these patients remains poor despite HAART. PMID- 12556692 TI - Effects of CCR5-Delta32 and CCR2-64I alleles on HIV-1 disease progression: the protection varies with duration of infection. AB - OBJECTIVE: To examine temporal variation in the effects of CCR5-Delta32 and CCR2 64I chemokine receptor gene polymorphisms on HIV-1 disease progression. DESIGN: Pooled analysis of individual patient data from 10 cohorts of HIV-1 seroconverters from the United States, Europe, and Australia. METHODS: We studied HIV-1 seroconverters of European (n = 1635) or African (n = 215) ancestry who had been genotyped for CCR5-Delta32 and CCR2-64I. We used Cox proportional hazards models with time-varying coefficients to determine whether the genetic protection against AIDS (1987 case definition) and death varied with time since seroconversion. RESULTS: Protection against AIDS conferred by CCR5-Delta32 held constant at a 31% (RH 0.69, 95% CI 0.54, 0.88) reduction in risk over the course of HIV-1 infection, whereas protection against death held constant at a 39% reduction in risk (RH 0.61, 95% CI 0.45, 0.88). When the period from AIDS to death was isolated, the survival benefit of CCR5-Delta32 diminished 2 years after AIDS. Protection against AIDS conferred by CCR2-64I was greatest early in the disease course. Compared with individuals without CCR5-Delta32 or CCR2-64I, individuals with one or two copies of CCR2-64I had a 58% lower risk of AIDS during the first 4 years after seroconversion (RH 0.42, 95% CI 0.23, 0.76), a 19% lower risk during the subsequent 4 years (RH 0.81, 95% CI 0.59, 1.12), and no significant protection thereafter. CONCLUSION: The protection against AIDS provided by CCR5-Delta32 is continuous during the course of infection. In contrast, the protection provided by CCR2-64I is greatest early in the course of infection. PMID- 12556693 TI - The long-term impact of HIV and orphanhood on the mortality and physical well being of children in rural Malawi. AB - OBJECTIVE: To assess the influence of maternal HIV status and orphanhood on child mortality and physical well-being. DESIGN: Retrospective cohort study with > 10 years of follow-up. METHODS: From population-based surveys in Karonga District, Malawi in the 1980s, 197 individuals were identified as HIV-positive. These individuals and 396 HIV-negative individuals matched for age and sex, and their spouses and offspring, were sought in 1998-2000. RESULTS: All but 11 of the index individuals were traced, identifying 2520 offspring; of these, 1106 offspring were included in analyses. Among those with HIV-positive mothers, mortality was 27% [95% confidence interval (CI), 17-38] in infants (1-30 days), 46% (95% CI, 34 58) in those under 5 years and 49% (95% CI, 38-61) in those under 10 years. The corresponding figures for those with HIV-negative mothers were 11% (95% CI, 8 13), 16% (95% CI, 13-19) and 17% (95% CI, 14-20). Death of HIV-positive mothers, but not of HIV-negative mothers or of fathers, was associated with increased child mortality. Among survivors who were still resident in the district, neither maternal HIV status nor orphanhood was associated with stunting, being wasted, or reported ill-health. CONCLUSIONS: Mortality in children under 5 years is much higher in children born to HIV-positive mothers than in those born to HIV negative mothers. With 10% of pregnant women HIV-positive, we estimate that approximately 18% of under-5 deaths in this population are attributable to HIV. Most of the excess is attributable to vertical transmission of HIV. Our findings suggest that, in terms of physical well-being, the extended family in this society has not discriminated against surviving children whose parents have been ill or have died as a result of HIV/AIDS. PMID- 12556694 TI - Evaluating two adjustment methods to extrapolate HIV prevalence from pregnant women to the general female population in sub-Saharan Africa. AB - OBJECTIVE: To evaluate two methods for estimating HIV prevalence among the general female population of reproductive age by adjusting data observed among antenatal clinic (ANC) attendees. METHODS: We adjusted the HIV prevalence among ANC attendees in Fort Portal (Uganda; 1994-1995), Mwanza municipality (Tanzania; 1990-1991), rural Mwanza (Tanzania; 1991-1993), Mposhi district (Zambia; 1994), Chelston (Lusaka, Zambia; 1994, 1996 and 1998) and Ndola (Zambia; 1998), using firstly a method that accounts for differences in age-specific fertility by HIV serostatus and secondly a method that accounts for differences in HIV prevalence by fertility risk category and parity. RESULTS: The non-adjusted HIV prevalence among ANC attendees underestimates the prevalence among the general female population by 8.0% in Chelston in 1998 and by between 20.7% and 31.9% in all other cases. The adjusted prevalence obtained using the first method underestimates the prevalence among the general female population by about 0.5% in Fort Portal and Mposhi; it overestimates that observed in Chelston in 1994 and 1996 by about 3.5%, and that observed in Ndola, urban Mwanza and rural Mwanza, by 6.5%, 10.6% and 12.8%, respectively. The second method (applied for only four sites) provides an overestimate of 7.0% in Chelston in 1994 and an underestimate of 3.8% and 2.1% in Ndola and rural Mwanza, respectively. Both adjustment methods overestimate the 1998 prevalence in Chelston, producing less accurate estimates than the non-adjusted data. CONCLUSIONS: The HIV prevalence among women in the general population could be estimated fairly accurately by these methods in settings with mature epidemics. PMID- 12556695 TI - Breastmilk RNA viral load in HIV-infected South African women: effects of subclinical mastitis and infant feeding. AB - OBJECTIVE: To investigate determinants of breastmilk RNA viral load among HIV infected South African women, with particular attention to infant feeding mode and subclinical mastitis. DESIGN: Observational, longitudinal study. METHODS: Information on current infant feeding practice and a spot milk sample from each breast were obtained from 145 HIV-infected lactating women at 1, 6 and 14 weeks postpartum. The sodium/potassium (Na+/K+) ratio in milk was taken as an indicator of subclinical mastitis. The association between milk RNA viral load and maternal and infant characteristics was investigated using uni- and multivariate models. RESULTS: Milk viral load was below the limit of detection of the HIV RNA assay (< 200 copies/ml) in 63/185 (34.1%), 73/193 (37.8%) and 68/160 (42.5%) of samples at 1, 6 and 14 weeks, respectively. Multivariate models predicted between 13 and 26% of variability in milk viral load in the first 14 weeks. Low blood CD4 cell count (< 200 x 10(6) cells/l) during pregnancy and raised milk Na+/K+ ratio were significantly associated with raised milk RNA viral load at all times, but there were no consistent associations between infant feeding mode and RNA viral load in milk. There was a non-significant trend for the six infants known to be infected postnatally, compared with the 88 infants who remained uninfected, to have been exposed to breastmilk of higher viral load at each time point. CONCLUSIONS: Breast milk HIV RNA viral load in the first 14 weeks of life varied; high levels were associated with subclinical mastitis and severe maternal immunosuppression. Multivariate models had limited predictive value for milk RNA viral load, illustrating the multiple contributors to viral load. PMID- 12556696 TI - Environmental-structural factors significantly associated with consistent condom use among female sex workers in the Dominican Republic. AB - OBJECTIVE: To examine the influence of environmental-structural factors in promoting consistent condom use (CCU) among female sex workers (FSW) and their regular paying partners in the Dominican Republic. METHODS: A cross-sectional survey was conducted with 288 FSW recruited from 41 sex establishments in Santo Domingo from March to June 1998. Sex workers were asked about their sexual behavior, self-efficacy at negotiating safe sex, perceived intimacy with their most recent regular paying partner, and the physical, social and policy environment of the establishment where they worked. Factor and reliability analysis were utilized to develop aggregate measures for self-efficacy (Cronbach's Alpha 0.60), intimacy (Cronbach's Alpha 0.80), and environmental structural support (Cronbach's Alpha 0.72). RESULTS: Controlling for sociodemographic characteristics of participants in multivariate analyses, environmental-structural support for condom use and HIV/sexually transmitted infection (STI) prevention was a significant predictor of CCU (OR 2.16; CI 1.18 3.97) among FSW and their regular paying partners. Safe sex self-efficacy (OR 2.80; CI 1.31-5.97) and low perceived intimacy with the most recent regular paying partner (OR 7.20; CI 3.49-14.83) were also significantly associated with CCU in multivariate analysis. CONCLUSION: Environmental-structural support for condom use and HIV/STI prevention is a significant predictor of CCU among FSW in the context of regular paying partnerships. Environmental-structural factors, in addition to relational and individual cognitive factors, should be assessed and addressed by behaviorally guided theory, research and interventions related to HIV/STI prevention and female sex work. PMID- 12556697 TI - Risk factors for Kaposi's sarcoma-associated herpesvirus infection among HIV-1 infected pregnant women in the USA. AB - OBJECTIVES: We sought to identify risk factors for infection with the Kaposi's Sarcoma-associated herpesvirus (KSHV) among pregnant women and to examine a reported association of KSHV with injecting drug use (IDU) and hepatitis C virus (HCV) infection. DESIGN: Cross-sectional evaluation of questionnaire data and KSHV and HCV seroprevalence in the Women and Infants Transmission Study. METHODS: In sera collected from HIV-1-infected pregnant women (n = 887) and, at age 12 months, their offspring (n = 900) at six sites in the USA and Puerto Rico, KSHV and HCV antibodies were detected with sensitive and specific enzyme immunoassays. Risk of KSHV was estimated by the unadjusted and adjusted odds ratio (OR(adj)) and 95% confidence interval (CI). The geographic referent sites were Chicago and Boston. RESULTS: Forty-seven (5.3%) of the women and three (0.3%) of the infants were KSHV seropositive. In univariate and multivariate analyses, KSHV in the women was associated with enrollment in Puerto Rico, Houston or Brooklyn (OR(adj), 4.3; 95% CI, 1.8-10.4) or Manhattan (OR(adj), 9.8; 95% CI, 3.7-25.6); non-completion of high school (OR(adj), 1.8; 95% CI, 0.9-3.4); the number of sexually transmitted diseases (OR(adj), 1.4; 95% CI, 1.0-1.9 per disease); and especially with IDU and HCV infection (OR(adj), 3.5; 95% CI, 1.5-7.9). CONCLUSIONS: Transmission of KSHV by blood inoculation may be highly inefficient, but our data support the hypothesis that it does occur. Large formal studies to evaluate whether KSHV transmission occurs via transfusion are needed to inform decisions regarding screening volunteer blood donors to protect the blood supply. PMID- 12556698 TI - Occurrence of Pneumocystis carinii in HIV-positive patients with suspected pulmonary tuberculosis in Ethiopia. AB - OBJECTIVE: To investigate the prevalence of Pneumocystis carinii in consecutive HIV-positive patients with suspected pulmonary tuberculosis (PTB) attending a university hospital in Ethiopia. METHODS: A PCR for P. carinii and an indirect immunoflorescence (IF) assay were performed on expectorated sputum samples from: 119 HIV-1-positive patients with negative smears and sputum cultures for Mycobacterium tuberculosis; 96 HIV-1-positive patients with culture-verified PTB; and 97 HIV-negative patients with negative mycobacterial cultures and 72 HIV negative patients with culture-verified PTB, serving as controls. Outcome of PCR and IF were compared with the chest radiographic (CXR) and initial clinical diagnosis. RESULTS: In the HIV+PTB- group, P. carinii was found in 10.9% by IF, 8.4% by single PCR (sPCR) and 30.3% by nested PCR (nPCR). In the HIV+PTB+ group, 3.1% were P. carinii positive by IF and sPCR and 13.5% by nPCR. All IF- and sPCR positive samples were nPCR positive. In the HIV-PTB+ and HIV-PTB- groups, 4.2% and 3.1% were nPCR positive, respectively. Six out of eight HIV+PTB- patients with CXR suggesting P. carinii pneumonia (PCP) were IF and/or nPCR positive for P. carinii. In the IF-positive and nested PCR-positive HIV+PTB- patients more than one-third were interpreted as PTB by CXR whereas only one patient was diagnosed with clinical PCP. CONCLUSIONS: P. carinii is prevalent in HIV-positive PTB suspects, suggesting that PCP may be an important, but not well recognized, differential diagnosis. Our findings have implications for treatment and primary prophylaxis for PCP in Ethiopia. PMID- 12556699 TI - CRF06-cpx is the predominant HIV-1 variant in AIDS patients from Ouagadougou, the capital city of Burkina Faso. PMID- 12556700 TI - Nelfinavir plasma concentrations in patients experiencing early failure with nelfinavir-containing triple combinations. PMID- 12556701 TI - Effect of co-formulated zidovudine, lamivudine and abacavir (Trizivir) on antiretroviral-naive patients presenting with advanced HIV-1 infection. PMID- 12556702 TI - Reply to "Different degree of immune recovery using antiretroviral regimens with protease inhibitors or non-nucleosides", by Barreiro et al. PMID- 12556703 TI - Resolution of azole-resistant oesophageal candidiasis in an AIDS patient after treatment with caspofungin. PMID- 12556704 TI - Salmonella typhimurium prostatic abscess: a rare cause of persisting dysuria in an AIDS patient. PMID- 12556705 TI - Mitochondrial DNA and sperm quality in patients under antiretroviral therapy. PMID- 12556706 TI - Efavirenz-induced psychosis leading to involuntary detention. PMID- 12556707 TI - Impact of ovarian cancer perceived by women. AB - Ovarian cancer may be particularly challenging for women, both physically and psychologically, because of the advanced nature of the disease at the time of diagnosis, the side effects of the disease, the repetitive cycles of aggressive therapy, and the perceived loss of femininity from the removal of reproductive organs. In addition, women with this disease rarely have an opportunity for cure. Most will face the very real possibility of dying. This qualitative study examined the perspectives of women living with ovarian cancer. Eighteen women participated in interviews, in which they described their experiences living with the disease. Women reported the myriad day-to-day changes in their lives, the major challenges they had to face, and the sources to which they turned for support. Implications for the practice of oncology nurses involve assessment, an understanding of the profound impact that this illness has on both the woman and her family, and the need to access a variety of supportive care programs. PMID- 12556708 TI - Women's experiences with recurrent ovarian cancer. AB - Most women with a diagnosis of ovarian cancer, because of the advanced nature of the disease at the time of diagnosis, will face the very real possibility of an early death. Thus the disease can have a significant impact on women who play a central role in the day-to-day management of family life, the nurture of children, and the care of extended family. This article reports the perspectives of women living with ovarian cancer regarding their experiences confronting recurrent disease. For this qualitative study 18 women were interviewed. Four primary themes emerged from the analysis of the verbatim interview transcripts: waiting for recurrence, facing the diagnosis of recurrence, managing treatment related concerns, and attempting to regain control. The practice implications for oncology nurses involve understanding the significant meaning associated with monitoring tumor markers and recurrent disease, assisting women in accessing information and support, and engaging in sensitive communication. PMID- 12556709 TI - Turning toward the psychosocial domain of oncology nursing: a main problem analysis in the Netherlands. AB - The purpose of this descriptive exploratory study by the Dutch Oncology Nursing Society was to describe the problem areas in the care of patients who are receiving chemotherapy as perceived by patients and professional caregivers as well as to explore differences between the perceptions of patients and professional caregivers. Three independent samples of 120 patients who are being treated with chemotherapy, 42 chemotherapy prescribing medical specialists, and 68 oncology nurses who are involved in the daily bedside care of the patients undergoing chemotherapy participated in the study. Patients and professional caregivers completed a 69- and a 58-item self-administered questionnaire, respectively. Results show that patients were satisfied with the medical and nursing care they received. Furthermore, results suggest that professional caregivers perceive the experience of patients with chemotherapy differently than patients, eg, patients perceive chemotherapy as reassuring that something is being done; whereas caregivers think that patients experience chemotherapy as something to fear or be anxious or uncertain about. It further seems that professional caregivers are more prone to act upon the medical-technical aspects of the treatment than to act on psychosocial sequelae of the treatment. PMID- 12556710 TI - Exploring and encouraging through social interaction: a qualitative study of nurses' participation in self-help groups for cancer patients. AB - Self-help groups are a growing phenomenon across national borders. Current sociologic empirical evidence shows that nurses and other healthcare professionals have become an integral part of self-help groups. The aim of the study is to describe and highlight the experiences of patients with cancer (n = 21) and oncology nurses (n = 12) with self-help groups. These experiences are drawn on to illustrate the characteristics of professional involvement in self help groups for patients with cancer. Data were obtained by individual qualitative interviews. The results show that the nurse functions as a social networker and uses her contextual competence by consciously encouraging relationships between fellow patients. Furthermore, the study illustrates that the nurse's involvement with self-help groups for patients with cancer serves as a complementary dimension to the traditional nursing discourse. It is concluded that when individualized care is supported through social practice and when personal issues are exchanged and negotiated, the nurse facilitates a milieu of togetherness in self-help groups for patients with cancer. The concept of self help groups is a valuable contribution to new theories and service development in psychosocial care and complies with the understanding of the postmodern individual, who viewed as primarily responsible for negotiating, socializing, and making his or her own decisions. PMID- 12556711 TI - Managing cancer side effects to improve quality of life: a cancer psychoeducation program. AB - This study aimed to develop and pilot test a community-based education program that addresses the prevention and management of the major side effects of cancer and its treatment according to a Patient Active Empowerment Model. This piloted program evaluated the health and quality of life outcomes for patients with cancer. The program was implemented through a half-day patient education conference. This conference included clinical information concerning disease and treatment-related side effects, an overview of treatment methods, information to help improve communication between the patient and healthcare team, education materials and experiences to empower patients and caregivers, and integration of specific mind-body techniques to strengthen key educational messages and reinforce the importance of side effect management. A total of 32 patients completed both the program and psychometric assessments on the day of the intervention and 30 days later. The participants were primarily white women with an average age of 57 years. Breast cancer was the most prevalent type of cancer experienced by the participants. The pilot results for this small sample showed that from baseline to follow-up evaluation, patients reported significant decreases in depressive symptoms and problems with work or other daily activities that resulted from emotional distress. Some improvements in health and well-being were greater for those who had not experienced fever or infection, those who had no children, and those who were working. Participants indicated that the program was useful in its ability to help them manage treatment side effects. Although the findings presented in this article are based only on a pilot program evaluation and a small sample, they do suggest that the program may be effective in educating patients about specific cancer side effects and empowering them to cope more effectively with their illness. Results showing that subgroups of individuals may have benefited more from the intervention provide important information about specific components of the program that may be particularly salient and potential changes that might be usefully implemented. PMID- 12556712 TI - Making the most of the moment: when a child's mother has breast cancer. AB - The demands and uncertainties associated with adjustments to chronic illness present challenges to maintaining a stable family life. Little has been reported about mother-child relationships within the cancer experience and even less about interventions that may be useful to help these dyads maintain stability. The focus of this article is a pilot study of an intervention addressing the mother child relationship of women with breast cancer and their school-aged children. A part of the data collection comprised interviews of mothers and fathers to assess their perceptions of the influence of the intervention on the quality of the mother-child relationships after the program. Using inductive content analysis, the fathers' and mothers' data were organized into categories, domains, and an explanatory construct, called making the most of the moment. The processes by which the intervention affected the mother-child relationships and implications for professionals who work with families are reported. PMID- 12556713 TI - Living with untreated localized prostate cancer: a qualitative analysis of patient narratives. AB - Few, if any, qualitative studies aimed at gaining an understanding of the experience of patients with prostate cancer have been done. The purpose of this study was to illuminate the meaning of being a patient living with untreated localized prostate cancer. Seven men with untreated localized prostate cancer were interviewed in their homes. The interviews were tape recorded and transcribed into text. The text was analyzed using a phenomenologic-hermeneutic approach inspired by Ricoeur's philosophy. The meaning of living with untreated localized prostate cancer could be interpreted as living life under a dark shadow. The disease was described as a threat to the patient's life. When living under this shadow, many of the men studied had an ambivalent wish both to share their experience with others and to be alone with their experiences of the disease. They believed that the disease had changed their lives, and their manhood was restricted by sexual dysfunctions and described as a burden. They used various coping strategies to manage this situation. Despite a positive relationship with their physicians, there is a risk that these patients will not be given the attention they need because of their good prognosis. PMID- 12556714 TI - Pain and hope in patients with cancer: a role for cognition. AB - The importance of hope in determining the adjustment of patients with cancer to their illness has been recognized. Stressful events such as pain and disease metastasis may have an impact on patients' hope levels. This study had three purposes: 1) to examine the effect of disease status on hope levels among patients with cancer who have pain; (2) to compare the level of hope between patients with cancer who have pain and those do not; and (3) to determine which dimensions of pain are associated with hope. Patients (n = 226) with various cancer diagnoses completed the Herth Hope Index. Disease status was measured by one objective indicator (disease stage) and one subjective indicator (perceived treatment effect). The Perceived Meanings of Cancer Pain Inventory was used to measure the cognitive dimension of pain, whereas pain intensity and relief were used to represent the sensory dimension of pain. The patients' disease stage did not affect their level of hope, but their perception of treatment effect was associated with this factor. No difference in level of hope was found between patients with pain and those without pain. For those with pain, the cognitive dimension of pain (meaning ascribed to pain) was significantly correlated with hope, whereas sensory dimensions (pain intensity and relief) showed no such correlation. The study results support the role of cognition in promoting the psychological well-being of patients with cancer. PMID- 12556715 TI - Patients with cancer and their close relatives: experiences with treatment, care, and support. AB - This article presents findings from a study of 536 patients with cancer and 473 of their close relatives from 15 different hospitals in Norway. The study assessed the satisfaction of both groups with the help and support they received from different sources such as family and friends, health personnel, and fellow patients. The results indicate that the patients received more support and information than their close relatives. The patients also were generally more satisfied with the support. Furthermore, the relatives seemed to underestimate the help and support provided to the patients. For both the patients and their close relatives, family and friends proved to be important sources of support. Regarding the public health services, almost 90% of the patients were very satisfied with the medical treatment. On the other hand, only an approximate 13% were very satisfied with information about alternative treatment and financial issues related to the disease. For the patients, the contact with fellow patients was highly valued. For most of the patients, this contact provided hope for the future, but it also had the potential for painful and sad feelings. The study was conducted in collaboration with the Norwegian Cancer Society to provide information that can be used for the guidance of health personnel and care of patients and their relatives. PMID- 12556716 TI - Improving the nursing management of patients in a hematology/oncology day unit: an action research project. AB - This article reports on an action research project conducted in one hematology oncology day unit in Queensland, Australia. The project consisted of three phases, each of which included observation, reflection, action, and evaluation. The first phase calculated the amount of time and the dedicated nursing hours required to administer the different chemotherapeutic protocols to patients in a number of hematology-oncology day units in Southeast Queensland. The second phase changed the patient booking and staff allocation systems following an evaluation of process and outcome measures and a patient survey. The patient survey measured both generic satisfaction and satisfaction with symptom management. The third phase introduced and evaluated the effectiveness of primary nursing and increased emphasis on patient education. The first two changes brought about by the action research process improved patient wait time, patient satisfaction, and symptom management. The changes implemented in phases 1 and 2 increased the amount of time available for nurses to engage in psychosocial care. The increased emphasis on education in phase 3 did not significantly improve patient knowledge related to self-care or appropriate health professional referral. To improve patient education, the development and implementation of a self-care education program is recommended. PMID- 12556717 TI - Malignant transformation of atypical oral lichen planus: a review of 32 cases. AB - AIMS: To assess the prevalence of malignant transformation of atypical lichen planus in our medium and describe the clinico-pathological features of the cases that had a premalignant potential. MATERIALS AND METHODS: we performed a retrospective analysis of the records of patients seen at the Stomatology Department of the School of Dentistry, University of Buenos Aires, during the period 1991-1997. A total of 719 patients (7.02%) had oral lichen planus (481 women and 238 men). From a clinico-pathological point of view, 228 (31.7%) were typical forms and 491 (68.3%) were atypical forms. In all of the cases, lichen planus was confirmed histologically and a clinical protocol was followed. RESULTS: Thirty-two (6.51%) patients were known to have developed malignant changes. In these cases the lesions had been diagnosed as atypical lichen planus at the outset. The most frequent clinical presentations were the keratotic (plaque-like) and erosive forms. Most of the lesions were on the tongue. The results support a premalignant potential for atypical lichen planus. CONCLUSION: Oral lichen planus is a precancerous condition. The atypical form, particularly in its keratotic and erosive presentations, has a premalignant potential in our medium. The most frequent localization is the tongue. PMID- 12556718 TI - A protocol for the evaluation and treatment of oral mucositis in patients with hematological malignancies. AB - A study is made to determine the true incidence of oral lesions attributable to hematological malignancies and their therapies, with the purpose of preventing, diagnosing and treating the oral manifestations of the hematological disorder and the situations of mucositis produced by the different chemo or radiotherapeutic regimens administered. A review was made of patients with hematological malignancies undergoing treatment or programmed for hematopoietic transplantation, with clinical assessment and microbiological culture or histological study where required. A protocol is developed in these patients for the prophylaxis, control and management of their oral lesions, thus contributing to improve the knowledge and treatment of patients with hematological malignancies. PMID- 12556719 TI - Oral manifestations of Langerhans cell histiocytosis. Case study of a two-year old boy. AB - Langerhans cell histiocytosis is a rare pathology. It may be a single system disease at a single site or at multiple sites, or a multisystem disease; it may evolve from one of these forms into another. Oral manifestations may be the first signs of the disease. Little is known as yet about its etiology. We review the nosological entity and present the case of a two-year-old boy suffering from the multisystem form, paying particular attention to the oral manifestations. Physical examination on admittance revealed skin lesions in the scalp, armpits and groins, hepatosplenomegaly and bilateral otorrhea. The diagnosis was multisystem Langerhans cell histiocytosis, affecting the skin, bone, liver and CNS. PMID- 12556720 TI - Human repercussions of foot and mouth disease and other similar viral diseases. AB - Foot and mouth disease is a frequent viral zoonosis in livestock that may occasionally also affect humans. Transmission to man usually occurs as a result of the consumption of unprocessed milk. The clinical manifestations include fever, headache, weakness, muscle pain, and the development of vesicles and ulcers throughout the oral mucosa. Vesicular stomatitis is another zoonosis similar to foot and mouth disease that can likewise affect humans with similar clinical manifestations, in which the presence of aphthae is highly suggestive. In turn, hand, foot and mouth disease and herpangina are two exclusively human diseases caused by different enteroviruses, with a special predilection for children under five years of age, and characterized by the presence of vesicles and ulcerations in the oral cavity. The present study provides a brief review of the salient characteristics of foot and mouth disease and of other similar viral diseases with which the differential diagnosis should be established. PMID- 12556721 TI - Lichenoid reaction caused by Clopidogrel, a new anti-platelet drug. AB - The case is described of a 71-year-old woman presenting with acute erosive lichen planus. The lesions appeared one week after starting treatment with clopidogrel (Plavix), a new antiplatelet drug, for the prevention of cerebral thrombosis. The patient presented erosive lesions on both jugal mucosal surfaces and the lateral sides of the tongue, associated with intense pain. The clinical diagnosis of erosive lichen planus induced by clopidogrel was confirmed by histological biopsy evaluation. Clopidogrel was replaced by aspirin, which was the antiplatelet drug previously used by the patient, and 0.1% triamcinolone acetonide in orabase was prescribed in combination with systemic corticoids. The erosive lesions and pain disappeared as a result, though asymptomatic reticular lesions typical of lichen planus persisted. PMID- 12556722 TI - Kindler syndrome: a clinical case. AB - Kindler's syndrome is a congenital disorder with simultaneous clinical characteristics of two congenital diseases: dystrophic epidermolysis bullosa and congenital poikiloderma. A review is made of the literature, with the presentation of the case of a 51-year-old woman who after a number of years of implant therapy for oral rehabilitation (total upper and partial lower arch edentulous status) developed xerostomia and a series of whitish and hyperkeratotic lesions associated with other atrophic erythematous lesions lining the entire oral mucosa. The clinical case and treatment are described, with the recommendation to ensure the follow-up of such patients, in order to avoid possible degenerative transformations of the disease, in view of its chronic nature. PMID- 12556723 TI - An epidemiological study of caries in a group of children with cerebral palsy. AB - The results of an epidemiological study of caries in a group of 103 children with cerebral palsy between 5 and 20 years of age and treated in Nen Deu Hospital (Barcelona, Spain) are presented. The CAOD (caries, absent teeth, filled teeth) and ICO indices (caries, filled teeth) were evaluated in the permanent and deciduous dentition, respectively. PMID- 12556724 TI - Coffin-Lowry syndrome: odontologic characteristics. Review of the literature and presentation of a clinical case. AB - A description is made of the general and odontologic characteristics of Coffin Lowry syndrome, with a review of the literature and the report of a clinical case. PMID- 12556725 TI - Lingual thyroid: a clinical case. AB - Lingual thyroid is an abnormal formation appearing as the result of a deficient descent during embryological development of the thyroid gland through the thyroglossal duct to its normal pretracheal location. The lesion consists of a tumor mass of thyroid tissue located at the base of the tongue, in the region of the foramen caecum linguae. The size can vary from a few millimeters to several centimeters in diameter. More than 400 cases of lingual thyroid have been documented in the literature to date. Lingual thyroid has been identified in 10% of the tongues examined in some autopsy series. Its identification is of great significance, since it may constitute the only functional thyroid tissue in the body, and may inadvertently be destroyed as a result of histological biopsy procedures. The present study presents a clinical case of lingual thyroid in a 17 year-old female. PMID- 12556726 TI - Distraction of oral scars contractures following caustic ingestion. A form of conservative treatment. AB - The chemical burns in the oral mucosa as consequence of accident of suicide attempt are relatively frequent. The evolution of the repairing process is conditioned by the quality and concentration of the caustic as by the location and time exposure of the affected region. At this report, we document a form of conservative treatment for the limitation of mouth opening, which the patient presented after swallowing a detergent acid. In order to obtain a minimal oral opening, a screwed cone, whose action mechanism was acting as a cradle between both jaws, was used. Once achieved a 10 mm opening, it was possible to use an external distractor to accomplish a continuous traction. PMID- 12556727 TI - Amyloidosis of the submaxillary gland. AB - Amyloidosis can be defined as an extracellular deposit of the amyloid fibrillar protein in one or more locations in the organism. Multiple clinical and biochemical forms of amyloid exist, and are characterized according to their fibrillar structural characteristics. The Red Congo stain presents green fluorescence when histologic cuts are observed by polarized light. The deposits of this substance can be focal in nearly any area of the organism, although the perivascular space tends to be affected. We present the case of a woman operated on for myeloma at the base of the cranium who showed no wider affectation at a later study. Fourteen years after the operation the patient showed a progressive growth of the submandibular gland that was diagnosed as a pathological amyloid deposit. PMID- 12556729 TI - [Extraction of impacted teeth]. PMID- 12556728 TI - Neck schwannomas. AB - Schwannomas are peripheral nerve tumours of nerve sheath origin. Twenty-five to 45 percent of extracranial schwannomas occur in the head and neck region. We present 2 cases of schwannomas that arise from the vagus and cervical plexus. These lesions are uncommon and most often present as asymptomatic solitary neck mass. Preoperative diagnosis can be difficult and conservative surgical excision remains the treatment of choice, often requiring sacrifice of a portion of the nerve. PMID- 12556730 TI - [Antibiotic prophilaxis of bacterial endocarditis]. PMID- 12556731 TI - Beyond serum creatinine: defining the patient with renal insufficiency and why? AB - Chronic kidney disease is the most important factor in predicting adverse short- and long-term outcomes after percutaneous coronary intervention. Most studies of cardiovascular outcomes have found that a break point for the development of radiocontrast nephropathy (RCN), later restenosis, recurrent myocardial infarction, congestive heart failure, and cardiovascular death, occurs below an estimated glomerular filtration rate (eGFR) of 60 mL/min/1.73 m2, which roughly corresponds to a serum creatinine (Cr) of > 1.5 mg/dL in the general population. Renal dysfunction is accurately recognized by calculating the eGFR from the age, serum creatinine, gender, race, and weight, and not from the serum creatinine alone. The pathogenesis of RCN goes beyond serum Cr and involves a unique vascular pathobiology that interrelates both the renal and cardiovascular disease outcomes. PMID- 12556732 TI - Radiocontrast nephropathy: identifying the high-risk patient and the implications of exacerbating renal function. AB - Radiographic procedures using contrast media are widely performed throughout the world. This necessitates physicians' awareness of radiocontrast nephropathy-the disorder that develops as a result of exposure to contrast agents. Although in the general population the risk of radiocontrast nephropathy is rather low, it may be very high in selected subsets of patients. This article focuses on the incidence, pathogenesis, risk factors, and prognosis of radiocontrast nephropathy and provides important insights on its prevention. PMID- 12556733 TI - A review of contemporary prevention strategies for radiocontrast nephropathy: a focus on fenoldopam and N-acetylcysteine. AB - The mechanism most likely responsible for the development of radiocontrast nephropathy (RCN) is contrast-induced renal tubular ischemia. At this time, intravenous hydration remains the mainstay for preventing RCN. The antihypertensive agent fenoldopam has been shown in a canine model, as well as in small, retrospective, prospective, and randomized human evaluations, to be effective for preventing RCN. In addition, studies have reported the ability of the free radical scavenger N-acetylcysteine (NAC) to prevent RCN. The clinical trial data for NAC, however, are not consistent regarding this effect, which, if present, appears to be modest and perhaps restricted to lower-risk clinical scenarios. PMID- 12556734 TI - Contemporary strategies to preserve renal function during cardiac and vascular surgery. AB - Mortality rates associated with perioperative acute renal failure (ARF) range from 60% to 90%. The major causes of ARF are prerenal factors that decrease renal blood flow; intrarenal factors that have a direct effect on tubules, interstitium, or glomeruli; and postrenal factors that obstruct urine outflow. Current strategies to provide perioperative renal protection include maintaining adequate renal O2 delivery, suppressing renovascular vasoconstriction, renovascular vasodilatation, maintaining tubular flow, decreasing renal cellular O2 consumption, and attenuating reperfusion injury. A study of patients undergoing elective repair of a thoracoabdominal aortic aneurysm (TAAA) found that the use of the selective dopamine-1 receptor agonist fenoldopam was associated with reductions in mortality, dialysis requirements, and lengths of stay in the hospital and intensive care unit. The study authors suggest that the improved patient outcomes and hospital-utilization data resulting from the use of fenoldopam were directly related to the protection of renal function during surgery and a reduction of postoperative renal complications. PMID- 12556735 TI - Improving perioperative outcomes in patients with end-stage heart failure. AB - In the United States alone, more than 4.5 million people are affected by heart failure, with more than 500,000 new cases diagnosed each year. Although cardiac transplantation remains the "gold-standard" surgical treatment for heart failure unresponsive to maximal medical therapy, the chronic shortage of donor hearts has necessitated clinical trials of other surgical options. Over the past two decades, research, technological progress, and extensive clinical experience have resulted in the application of ventricular assist device (VAD) technology to a broader population of heart-failure patients, as these devices have proven to be viable therapeutic alternatives for therapy of end-stage heart failure. All patients undergoing cardiac transplantation or VAD insertion have multiorgan dysfunction as a result of irreversible, severe ventricular dysfunction resulting in low cardiac output. Recently, fenoldopam has been described as a vasodilator that might be useful in patients with decompensated heart failure, particularly in the perioperative setting. As a selective dopamine-1 receptor agonist, fenoldopam causes vasodilation in the systemic, renal, mesenteric, coronary, and pulmonary vasculature. Potentially, the pharmacologic properties of fenoldopam could be successfully exploited in patients undergoing medical or surgical treatment of end-stage heart failure. Controlled randomized trials are needed to demonstrate improvement in cardiopulmonary or renal outcomes in such patients. PMID- 12556736 TI - The role of the DA1 receptor agonist fenoldopam in the management of critically ill, transplant, and hypertensive patients. AB - Fenoldopam, a selective agonist of dopamine-1 receptors, is a regional and systemic vasodilator. In randomized, controlled clinical trials, fenoldopam has been found to preserve renal function in situations of potential renal ischemia, such as during radiocontrast administration, cardiac and peripheral vascular surgery, liver transplantation, and treatment of severe hypertension. Fenoldopam lowers blood pressure in patients with hypertension, but has little or no effect on blood pressure in those who are normotensive. The role of fenoldopam in managing critically ill, transplant, and hypertensive patients is reviewed in this article. PMID- 12556737 TI - Clinical diagnosis and prognosis of acute limb ischemia. AB - Acute limb ischemia (ALI) is a serious condition that can threaten the limb's viability and the patient's life. It is usually caused by atherosclerotic disease but can also arise from nonatherosclerotic causes. It has numerous clinical manifestations, the most common of which is pain. Proper diagnosis involves history, physical examination, and techniques such as Doppler assessment, contrast-enhanced magnetic resonance angiography, or catheter-based angiography. Differential diagnosis is important, as many other conditions can mimic ALI. Ischemia severity can be classified into three levels: viable; threatened, either marginally or immediately; and irreversible. Treatment options include surgical and less invasive endovascular techniques. Prognosis can be excellent, and early diagnosis and prompt initiation of therapy are essential to achieving a good outcome. PMID- 12556738 TI - Comparison of surgical and thrombolytic treatment of peripheral arterial disease. AB - Acute occlusion of a peripheral artery is a catastrophic event. Whether resulting from in situ thrombosis of a native artery, a bypass graft, or embolization, acute limb ischemia threatens both the patient's limb and life. Traditionally, open surgical intervention has been the "gold standard" for treatment of these patients. However, the multiplicity and complexity of medical comorbidities account for high rates of perioperative morbidity and mortality. Thus, a minimally invasive alternative to open surgery is desirable, provided that the rate of limb salvage remains similar and other untoward events are infrequent. Catheter-directed thrombolytic therapy has been studied in this regard, offering the potential to restore arterial perfusion without the need for open surgery in many cases. In addition, thrombolysis can clear thrombus from small arteries that are inaccessible to a balloon catheter. Lastly, successful thrombolysis may unmask the lesion responsible for the occlusion and allow a directed, sometimes less invasive treatment. Thrombolysis has been criticized, however, on the basis of associated hemorrhagic complications, a slow rate of thrombus dissolution, and a higher risk of rethrombosis. This article explores the available data and, in this manner, provides an analysis of open surgery and thrombolytic therapy as initial interventions in patients with lower limb ischemia. PMID- 12556739 TI - Comparison of safety and efficacy of the various thrombolytic agents. AB - Thrombolytic agents are in widespread use for the dissolution of arterial and venous pathologic thrombi. Clinical settings where thrombolysis has played an important role include the acute coronary syndromes, peripheral arterial occlusion, ischemic stroke, deep venous thrombosis, and pulmonary embolism. Thrombolytic agents have been successfully employed in each of these areas, achieving dissolution of the occluding thrombus, reconstitution of blood flow, and improvement in the status of the tissue bed supplied or drained by the involved vascular segment. All clinically available thrombolytic agents act through cleavage of the plasminogen molecule to its active form, plasmin. Despite this similar mechanism of action, the thrombolytic agents differ in several biochemical parameters, including fibrin specificity, fibrin affinity, and relative resistance to inactivating factors in the plasma. Whether these differences account for significant differences in clinical outcome is a matter of some dispute. It is quite possible that in vitro biochemical differences do not have meaningful clinical correlates. However, there exists some evidence to suggest that differences in the risk of distant hemorrhage, idiosyncratic reactions, and the rapidity of clot dissolution do exist. An ideal agent for peripheral vascular thrombolysis would be one that was specific in its actions at the site of pathologic thrombi yet left the important and desirable pathologic thrombi that seal vascular defects unscathed. Although such an agent has not yet been identified, an understanding of the mechanism of action and principles underlying pharmacologic thrombolysis provides the necessary foundation of knowledge to choose a particular thrombolytic agent for a given clinical scenario. PMID- 12556741 TI - Present-day thrombolytic therapy: therapeutic agents--pharmacokinetics and pharmacodynamics. AB - Various thrombolytic agents have been studied as activators of the plasminogen plasmin system for thrombolysis of thrombus formation. They include streptokinase, urokinase, tissue plasminogen activators, single-chain urokinase plasminogen activator, and anisoylated or acylated plasminogen-streptokinase activator complex (APSAC), only some of which are commercially available. All thrombolytic agents, including APSAC (not commercially available), recombinant tissue plasminogen activator, and prourokinase, generate great quantities of degradation products of fibrinogen or fibrin. All of the second-generation thrombolytic agents induce systemic activation of the entire fibrinolytic system, and none are capable of specifically activating the fibrinolytic system at the site of thrombus formation. The most systemically active agent known at the present time is APSAC. Trials show that bleeding occurs as frequently with the second-generation agents as with the older agents, and further studies may even find that the newer agents are associated with more bleeding than urokinase and streptokinase have been. With knowledge of the properties of the various thrombolytic agents available today, the physician can intelligently select the optimal agent for a given patient problem. PMID- 12556742 TI - Mechanical thrombectomy devices for the treatment of peripheral arterial occlusions. AB - The changing landscape of thrombolytic drugs used for treating peripheral occlusive disease has spurred increasing use of mechanical thrombectomy devices (MTDs). These devices comprise a variety of tools intended to remove, fragment, or disperse thrombus in veins, arteries, or bypass grafts. Although most devices are U.S. Food and Drug Administration approved for use in occluded hemodialysis access grafts, only one, the AngioJet device, is currently approved for use in peripheral arterial applications. Nevertheless, most devices have been used for this off-label application, and clinical centers have published their results. This article reviews the distinguishing features of current mechanical thrombectomy devices, as well as the currently available literature reporting on their use in peripheral arterial occlusions. PMID- 12556743 TI - Thrombolytic therapy for acute deep vein thrombosis and the venous registry. AB - Randomized clinical trials have defined anticoagulation with unfractionated or low molecular weight heparin followed by warfarin as standard therapy for acute deep venous thrombosis (DVT). Such treatment is highly effective in preventing recurrent venous thromboembolism, but provides imperfect protection against development of the postthrombotic syndrome. By restoring venous patency and preserving valvular function, catheter-directed thrombolytic therapy potentially affords an improved long-term outcome in selected patients with DVT. A national venous registry, compiling data from 63 participating centers, was established to collect data regarding the technical details of the procedure and early outcome. Data from the registry have established the optimal technical approach and patient population. An antegrade catheter-directed approach using urokinase in patients with acute iliofemoral DVT of less than 10 days duration and no prior history of DVT may achieve complete lysis in 65% of patients. Analysis of the clinical outcome is pending, but early results suggest improved valve function and fewer symptoms at 1 year in patients with complete thrombolysis. These promising data should serve as the basis for future randomized trials of catheter directed thrombolysis for the treatment of acute DVT. PMID- 12556744 TI - Quality-of-life improvement using thrombolytic therapy for iliofemoral deep venous thrombosis. AB - Patients with iliofemoral deep venous thrombosis suffer the most severe postthrombotic morbidity. Techniques that effectively remove thrombus from the venous system eliminate venous obstruction and potentially preserve valvular function. This will likely reduce or avoid the postthrombotic syndrome and improve long-term quality of life. To evaluate whether catheter-directed thrombolysis is associated with improved quality of life compared with anticoagulation alone and whether outcome in the thrombolysis group is related to lytic success, 98 patients with iliofemoral deep venous thrombosis who were treated at least 6 months earlier were identified and queried with a validated health-related quality-of-life questionnaire. Sixty-eight patients were identified through the Venous Registry (a national, multicenter venous registry) and were treated with catheter-directed thrombolysis with urokinase, and 30 patients were identified by means of medical record review and were treated with anticoagulation alone. All patients were candidates for thrombolysis; however, the treatment decision was made according to physician preference. The two treatment groups did not differ significantly in average time between the reference hospitalization and first contact. No difference was found in physical functioning and well-being between the groups before the development of deep venous thrombosis. Following treatment, patients receiving catheter-directed thrombolysis reported better overall physical functioning, less stigma, less health distress, and fewer postthrombotic symptoms compared to those patients treated with anticoagulation alone. Within the thrombolysis group, successful lysis correlated with health-related quality of life. Catheter-directed thrombolysis for the management of patients with iliofemoral deep venous thrombosis significantly improves health-related quality of life compared to similar patients treated with anticoagulation alone. Improved quality of life is related to successful thrombolysis. These data offer a compelling argument for a prospective randomized study. PMID- 12556745 TI - Pulmonary embolism. AB - The natural history of pulmonary embolism (PE) is incompletely characterized, because most episodes of PE go undetected, the clinical presentation mimics so many other common and uncommon diseases, the sensitivity and specificity of the diagnostic tests are poorly defined, and even detection at autopsy is difficult and requires close examination of the pulmonary arteries. Yet PE is a significant cause of morbidity and mortality in the hospitalized patient, and one reason for its extremely high incidence is the failure of physicians to provide adequate prophylaxis to patients who are at risk of developing venous thromboembolism. The mortality rate for PE is less than 8% when the condition is recognized and treated correctly but approximately 30% when untreated. Pulmonary arteriography is still the gold standard in diagnosing pulmonary emboli, but several other imaging modalities have been used to diagnose pulmonary emboli in recent years, including transthoracic and transesophageal echocardiography, magnetic resonance angiography, spiral computerized tomography, and ventilation-perfusion lung scanning. The treatment modality chosen depends directly on the clinical presentation of the patient. Low molecular weight heparin may be equal or superior in efficacy to unfractionated heparin for the treatment of deep venous thrombosis and PE. Thrombolytic therapy can be considered for patients with hemodynamic instability, those with right ventricular dysfunction, and young patients with a massive PE despite a normal right ventricle on echocardiography. In those patients who cannot receive anticoagulation therapy or thrombolysis, or who remain at high risk, an inferior vena cava filter should be placed. PMID- 12556746 TI - Axillary-subclavian venous thrombosis. AB - Axillary-subclavian venous thrombosis (ASVT) is classified as either primary, which is often associated with active use of the upper extremities and underlying anatomic abnormalities, or secondary to recognized thrombotic risk factors. Although there is a risk of pulmonary embolism with both, late outcome varies with etiology. Primary ASVT occurs in young healthy patients, and up to 80% may have significant chronic symptoms. Secondary ASVT occurs in older patients with medical comorbidities, high mortality rates, and few late manifestations. Although anticoagulation is appropriate for most patients with secondary ASVT, a multidisciplinary approach including catheter-directed thrombolysis, thoracic outlet decompression, and correction of intrinsic venous lesions more effectively prevents late symptoms in selected patients with primary ASVT. PMID- 12556747 TI - Thrombolysis for the treatment of thrombosed hemodialysis access grafts. AB - Maintaining the patency of hemodialysis access grafts remains problematic. It is best to recognize the failing graft prior to its thrombosis by noting an increase in recirculation, decreased flow (as measured by a Transonics device), changes in Doppler ultrasound findings, elevation of venous pressures, or swelling of the arm. If a failing graft is suspected, an angiogram should be performed to evaluate the graft. If a problem is identified it should be corrected. If it is a graft thrombosis, it can be opened using percutaneous techniques. Percutaneous declotting has been evolving since its introduction in the early 1980s. At first, a low-dose thrombolytic infusion through a single catheter was used. Crossing catheters with a higher-dose infusion was then introduced. Finally, pharmacomechanical thrombolysis, which used crossing catheters and a pulse-spray technique, became popular. Several mechanical devices have proven to be efficacious as well. In 1997, we described the "lyse-and-wait" technique. We believe "lyse and wait" to be a simpler and quicker technique, and its initial success has been similar to that for the previously described techniques. After the graft is successfully declotted, the arterial plug must be mobilized and the stenotic lesion must be addressed either by angioplasty, stent placement, surgery, or any combination of these interventions. PMID- 12556748 TI - Interventional stroke therapy: the potential benefit of direct intra-arterial infusion. AB - The goal of acute stroke therapy is preservation of the ischemic penumbra, the geographic area of relatively less ischemia surrounding the profoundly ischemic center. Emergent intravenous revascularization is currently the only U.S. Food and Drug Administration-approved therapy, but intra-arterial revascularization has also been shown to improve outcome significantly. Intravenous therapy has significant limitations: the time window for therapy is short, the large dose of fibrinolytic agent required may increase the risk of hemorrhagic complications, and questions have been raised regarding efficacy in patients with large vessel occlusions. Intra-arterial delivery of the lytic agent can address these issues, and direct intracranial intra-arterial fibrinolysis has been shown to be effective. Thrombolytic agents differ in stability, half-life, and fibrin selectivity, but effective therapies for acute stroke therapy must provide one common denominator: restoration of flow. It now appears certain that direct intra arterial delivery of fibrinolytic agent improves the efficacy of lysis of bulk thrombus. The Interventional Stroke Therapy Outcomes Registry is acquiring data on all forms of interventional stroke therapy from around the nation as well as outside the United States to identify best practices in the treatment of acute stroke. PMID- 12556749 TI - Thrombolytic solutions for vascular thrombotic events: summary and conclusions. PMID- 12556750 TI - Pulmonary vein isolation for atrial fibrillation. AB - Nonpharmacological treatment for atrial fibrillation (AF) has evolved dramatically over the past 20 years. Pulmonary vein isolation, a catheter ablation technique developed in the last decade, prevents focal triggers in the pulmonary veins from initiating episodes of AF. Although the procedure initially involved focal ablation with a catheter directly in the pulmonary veins, investigators subsequently found that isolating the pulmonary veins by applying ablation energy at their junction with the left atrium is more effective. After the procedure is performed once or twice, it is 70% to 80% successful in preventing recurrence of AF episodes in the first year of follow-up. The pulmonary vein isolation procedure is most suitable for patients whose recurring symptomatic episodes of AF have not been suppressed by antiarrhythmic drugs or who do not wish to take long-term antiarrhythmic or anticoagulation medications. The procedure is more successful in patients with intermittent (paroxysmal) AF than in those with long-standing, continuous (chronic) AF. PMID- 12556751 TI - Current anticoagulation options in percutaneous intervention: designing patient specific strategies. AB - Anticoagulation during percutaneous coronary intervention is critical to prevent abrupt and subacute closure. Although heparin has been the primary anticoagulant used for this purpose, a number of new drugs are now available. Low molecular weight heparin (LMWH) offers some advantages over unfractionated heparin, and clinical trials have shown its superiority. However, the longer half-life and lack of monitoring of LMWH make its use more difficult. The direct thrombin inhibitors also have been shown to have advantages in the treatment of patients with heparin-induced thrombocytopenia. PMID- 12556752 TI - The angiotensin II receptor blockers: opportunities across the spectrum of cardiovascular disease. AB - Angiotensin receptor blockers are a new class of agents that have made a major contribution to the treatment of hypertension. These agents effectively reduce blood pressure and are well tolerated. Other clinical trials have focused, however, on the much wider use of angiotensin receptor blockers in conditions such as congestive heart failure, postmyocardial infarction management, and diabetic nephropathy. Recent studies have provided evidence that these agents might confer target organ protection in hypertension that is equal to, and possibly better than, the benefits provided by conventional antihypertensive agents. Moreover, there is now little doubt that these drugs are effective alternatives to ACE inhibitors in heart failure and will become treatments of choice for patients with type 2 diabetes and nephropathy. Cardiovascular study outcomes have still not determined, however, whether high-risk patients would do better on angiotensin receptor blockers or angiotensin converting enzyme (ACE) inhibitors or a combination of both, except in cases of intolerance to ACE inhibitors. PMID- 12556754 TI - Choice for stroke prevention. PMID- 12556753 TI - Antiphospholipid syndrome with renal artery embolism: case report. AB - Antiphospholipid syndrome is characterized by venous and arterial thrombosis. Antiphospholipid antibodies are essential to making the diagnosis. It may be a primary disorder or it may be secondary to a connective tissue disorder. Cardiac manifestations of this syndrome include both thrombotic and degenerative valvular disease. Systemic or pulmonary embolism, as well as intravascular thrombosis, is a significant cause of morbidity and mortality. We present a case of renal infarction in a woman with polymyalgia rheumatica and a positive test for antiphospholipid antibody. PMID- 12556755 TI - Immediate repair versus surveillance of small abdominal aortic aneurysms. PMID- 12556756 TI - Coronary artery disease. Prevention with statin and niacin. PMID- 12556757 TI - Congestive heart failure. Doppler imaging to predict beta-blocker performance. PMID- 12556759 TI - Uncover hidden trauma. PMID- 12556762 TI - Capnography in EMS. A powerful way to objectively monitor ventilatory status. PMID- 12556763 TI - 2003 resource directory. The place to go when you need to know. PMID- 12556764 TI - The silent killer: recognizing & treating carbon monoxide poisoning. PMID- 12556766 TI - GPS, computers, ambulances & getting lost. What do they have in common? PMID- 12556767 TI - Smarten up: don't take chances with computer & network security. PMID- 12556769 TI - Straight shot: use of restraints, part 2. PMID- 12556772 TI - The capnography revolution begins. PMID- 12556774 TI - Endoscopic sphincterotomy and endoscopic papillary balloon dilatation for bile duct stones: A prospective randomized controlled multicenter trial. AB - BACKGROUND: Endoscopic papillary balloon dilatation may be an alternative to endoscopic sphincterotomy in the treatment of bile duct stones. However, there is a controversy as to the effectiveness and safety of endoscopic papillary balloon dilatation. METHODS: Two hundred eighty-two patients with bile duct stones were enrolled and randomized to an endoscopic sphincterotomy or endoscopic papillary balloon dilatation group. The success rate for duct clearance as well as the frequency and types of complications were evaluated prospectively. Endoscopic sphincterotomy was performed in a standard manner. Endoscopic papillary balloon dilatation was carried out with gradual inflation of a 4-, 6-, or 8-mm diameter balloon. RESULTS: Complete duct clearance was achieved in 100% in the endoscopic sphincterotomy group and 99.3% in the endoscopic papillary balloon dilatation group (not significant). Complications occurred in 11.8% of patients in the endoscopic sphincterotomy group and 14.5% of those in the endoscopic papillary balloon dilatation group (not significant). No complication was severe; there was no mortality. The frequency of acute pancreatitis was higher in the endoscopic papillary balloon dilatation group than the endoscopic sphincterotomy group (respectively, 10.9% vs. 2.8%; p < 0.045). Hemorrhage occurred only in the endoscopic sphincterotomy group. CONCLUSIONS: Endoscopic sphincterotomy and endoscopic papillary balloon dilatation were approximately equal in terms of successful clearance of bile duct stones. They were also similar with respect to overall complications. Endoscopic papillary balloon dilatation is an alternative to endoscopic sphincterotomy as a treatment of bile duct stones. PMID- 12556775 TI - Biliary sphincterotomy plus dilation with a large balloon for bile duct stones that are difficult to extract. AB - BACKGROUND: Bile duct stones are still present in 10% to 15% of patients after the application of conventional endoscopic extraction techniques and require additional procedures for duct clearance. In the vast majority of these cases, there are 2 main problems: large stone size (>15 mm) and tapering of distal bile duct. METHODS: Fifty-eight patients in whom endoscopic sphincterotomy and standard basket/balloon extraction were unsuccessful in the removal of bile duct stones underwent dilation with a 10- to 20-mm diameter (esophageal/pyloric type) balloon at the same session. In 18 patients with tapered distal bile ducts (Group 1), 12- to 18-mm diameter balloon catheters were used to enlarge the orifice. In 40 patients with square, barrel shaped and/or large (>15mm) stones (Group 2), the sphincterotomy orifice was enlarged with 15- to 20-mm diameter balloon catheters. After dilatation, standard basket/balloon extraction techniques were used to remove the stone(s). RESULTS: Stone clearance was successful in 16 patients (89%) in Group 1 and 35 (95%) in Group 2. Complications occurred in 9 (15.5%) patients. CONCLUSION: Dilation with a large-diameter balloon after endoscopic sphincterotomy is a useful alternative technique in patients with bile duct stones that are difficult to remove with standard methods. PMID- 12556777 TI - Esophageal stenosis after endoscopic mucosal resection of superficial esophageal lesions. AB - BACKGROUND: Although bleeding and perforation are generally recognized major complications of endoscopic mucosal resection, esophageal stricture after endoscopic mucosal resection has not been well studied. Factors associated with the occurrence and severity of esophageal stenosis after endoscopic mucosal resection were investigated. METHODS: Two hundred sixteen superficial esophageal lesions in 137 consecutive patients who underwent endoscopic mucosal resection from February 1993 through March 2001 were retrospectively studied. The circumferential extent of the mucosal defect after endoscopic mucosal resection was classified into 4 groups: under one fourth, one fourth to one half, one half to three fourths, and over three fourths. The longitudinal length of the mucosal defect was also evaluated. Stenosis was diagnosed when a standard endoscope (11 mm diameter) could not be passed through the stricture. RESULTS: Esophageal stenosis developed after endoscopic mucosal resection of 13 lesions (6.0%). In all these cases endoscopic mucosal resection resulted in a mucosal defect that involved over three fourths of the luminal circumference. In the subgroup of patients with mucosal defects involving over three fourths of the circumference, those with a mucosal defect over 30 mm long required more frequent balloon dilatation (mean 8 [4.3] times) and the stenosis was of longer duration (mean 16 [17.7] months) than those with defects 30 mm or less in length (respectively, 1 [0.6] times and 2 [1.9] months). CONCLUSIONS: A circumferential mucosal defect involving over three fourths the circumference of the esophagus after endoscopic mucosal resection was significantly associated with the subsequent development of esophageal stenosis. In addition, mucosal defects longer than 30 mm were associated with greater severity of stenosis. When endoscopic mucosal resection is performed for superficial esophageal lesions, removal of excess mucosa should be avoided. PMID- 12556776 TI - Cost-effectiveness analysis of high-dose omeprazole infusion as adjuvant therapy to endoscopic treatment of bleeding peptic ulcer. AB - BACKGROUND: Intravenous administration of proton pump inhibitors after endoscopic treatment of bleeding peptic ulcers has been shown to decrease the rate of recurrent bleeding and the need for subsequent surgery. Yet there is a relative lack of formal assessment of this practice. The aim of this study was to examine the cost-effectiveness of this therapy by using standard pharmacoeconomic methods. METHODS: The present study was performed in conjunction with a randomized controlled clinical trial that included 232 patients who received either omeprazole (80 mg intravenous bolus followed by infusion at 8 mg/hour for 72 hours) or placebo after hemostasis was achieved endoscopically. A cost effectiveness analysis was performed to evaluate the different outcomes of the trial. All related direct medical costs were identified from patient records. Cost-effectiveness ratios were calculated. RESULTS: Analysis by the Kolmogorov Smirnov test showed that the direct medical cost in the omeprazole group was lower than that for the placebo group. Cost-effectiveness ratios for omeprazole and placebo groups were, respectively, HK$ 28,764 (US$ 3688) and HK$ 36,992 (US$ 4743) in averting one episode of recurrent bleeding in one patient after initial hemostasis was achieved endoscopically. CONCLUSIONS: Intravenous administration of high-dose omeprazole appears to be a cost-effective therapy in reducing the recurrence of bleeding and need for surgery in patients with active bleeding ulcer after initial hemostasis is obtained endoscopically. PMID- 12556778 TI - Adequacy of mucosal sampling with the "two-bite" forceps technique: a prospective, randomized, blinded study. AB - BACKGROUND: Although the "two-bite" technique (obtaining 2 mucosal biopsy specimens with a single passage of the forceps) is used routinely, the adequacy of the second biopsy for histopathologic examination has not been evaluated. Our hypothesis was that the second sample will be inadequate for this purpose. The aim of this study was to prospectively assess the adequacy of mucosal biopsy specimens obtained with 3 different types of forceps with the two-bite technique. METHODS: Patients undergoing diagnostic EGD were prospectively enrolled in the study. In each patient, mucosal biopsy specimens were obtained as follows: 4 specimens with the two-bite technique in 2 separate passages of the forceps (one from the esophagus followed by one from the stomach, and one from the stomach followed by one from the esophagus, the order being chosen randomly) and 2 single bite specimens from the same anatomic locations (esophagus and stomach). Each patient underwent the same biopsy sampling sequence with 3 different forceps. An experienced pathologist blinded to the sequence and technique of obtaining the samples and the forceps used evaluated the specimens for number submitted, integrity, and adequacy for histopathologic diagnosis and depth of the sample as assessed by the presence or absence of muscularis mucosae. RESULTS: A total of 288 mucosal samples were obtained from 16 patients. Of these, 192 were taken by using the two-bite technique. Thirty-five (18.2%) samples were missing when the two-bite technique was used compared with only 2 (2.1%) when the single bite technique was used (p < 0.05). Irrespective of the location from which the first mucosal sample was taken, a significantly greater number of first samples were lost (25%) compared with second samples (11.5%) (p < 0.05). The forceps without a spike was associated with significantly more missing samples than the spiked forceps (28.1% vs. 13.3%; p = 0.01). At histopathologic evaluation, there were no significant differences between first and second samples nor differences between samples taken with the two-bite and the single-bite techniques with regard to adequacy, integrity, and depth. With respect to histopathologic evaluation, there were no differences among the 3 types of forceps used in the study. CONCLUSIONS: Although the second mucosal sample obtained with the two-bite technique is adequate for histopathologic purposes, there is a significant risk of losing samples (the first one in particular) with this technique, and thus an increase in the probability of sampling error. This may be particularly true for forceps without a spike. PMID- 12556779 TI - Hydrogen breath test as an indicator of the quality of colonic preparation for colonoscopy. AB - BACKGROUND: Breath hydrogen levels after ingestion of polyethylene glycol were evaluated as a method of predicting the quality of colonic preparation. METHODS: One hundred patients undergoing nonemergency colonoscopy were recruited for this study. After fasting overnight, they were instructed to ingest a polyethylene glycol solution containing 12 g lactulose at a rate of 50 mL every 5 minutes for 2 hours. During ingestion of the polyethylene glycol solution, breath samples were taken at 15-minute intervals for 240 minutes and breath hydrogen concentration was measured. RESULTS: The preparation for colonoscopy was judged to be poor in 18% and adequate in 82%. The breath hydrogen levels over 90 minutes were significantly higher in the poor group than in the adequate group. In all patients with a breath hydrogen level less than 10 parts per million at 240 minutes, the preparation was adequate. Conversely, all patients with a poor preparation had a breath hydrogen level of more than 10 ppm at 240 minutes. CONCLUSIONS: The hydrogen breath test effectively predicts adequacy of colonic preparation. PMID- 12556780 TI - Plastic or metal stents for malignant stricture of the common bile duct? Results of a randomized prospective study. AB - BACKGROUND: The systematic use of metal stents to treat biliary obstruction is restricted by high cost compared with plastic stents. The aims of this study were to compare cost and efficacy of plastic stents and metal stents in the treatment of patients with malignant common bile duct strictures and to define factors that predict survival of these patients. METHODS: One hundred eighteen patients (mean age 75 years) with malignant strictures of the common bile duct were randomized to placement of a plastic stent or metal stent. Comparisons were made with the Mann-Whitney or chi-square test as indicated; survival rates were compared with a Cox proportional hazards model. RESULTS: There was no significant difference in survival between the two groups. Time to first obstruction was longer for patients in the metal stent group (metal stent, median not reached vs. plastic stent, 5 months; p = 0.007). The number of additional days of hospitalization, days of antibiotic therapy, and the numbers of ERCPs and transabdominal US procedures was significantly higher in the plastic stent group. After multivariate analysis, only the presence of liver metastases was independently related to survival (p < 0.0005; OR = 2.25). This variable defined a group with a shorter survival. Median survival of patients with hepatic metastasis at diagnosis was 2.7 months compared with 5.3 months for patients without liver metastasis; in the latter group, the overall cost associated with metal stents was lower than for plastic stents. CONCLUSIONS: Metal stent placement is the most effective treatment of inoperable malignant common bile duct stricture. Placement of a metal stent is cost effective in patients without hepatic metastases, whereas a plastic stent should be placed in patients with spread of the tumor to the liver. PMID- 12556781 TI - Effectiveness of open-access endoscopy in routine primary-care practice. AB - BACKGROUND: Direct referral of patients for endoscopic procedures without prior consultation (open-access endoscopy) has become commonplace. However, the effect of open-access endoscopy on the care of patients in routine clinical practice has not been studied. METHODS: The impact of open-access endoscopy was examined in 168 consecutive patients referred from 8 primary-care practices to our tertiary hospital-based endoscopy center. The effectiveness of open-access endoscopy was assessed by review of office medical records at the primary-care practice sites for a minimum follow-up period of 6 months. Outcome measures evaluated included postprocedure communication between primary-care physician and patient, primary care physician adherence to postprocedure recommendations, and the need for subsequent diagnostic evaluation and/or consultation. RESULTS: The mean age of the 168 patients was 60 years; 56 (33%) underwent EGD and 112 (67%) had colonoscopy; 65% were from hospital-based practices and 35% from community practices. The indication(s) for 77% of the procedures met American Society for Gastrointestinal Endoscopy guidelines for the appropriate use of endoscopy. An office follow-up was noted for 82% of patients after the open-access procedure. Discussion of results was documented in 61% of the patient charts. Compliance with diagnostic and therapeutic recommendations was documented, respectively, in 75% and 90% of patient charts. A follow-up GI consultation was requested for only 7% of the patients. CONCLUSION: Open-access endoscopy in the primary-care setting is effective to the extent that subsequent GI consultations are rare and the level of compliance with endoscopist recommendations is high. However, documentation of communication of the results of endoscopy with the patient can be improved. PMID- 12556782 TI - Duodenal histology in patients with celiac disease after treatment with a gluten free diet. AB - BACKGROUND: The diagnosis of celiac disease requires characteristic histopathologic changes in an intestinal biopsy with clinical improvement in response to a gluten-free diet. Endoscopy with procurement of biopsy specimens is often performed to document response to the diet, but there are little data on the appearance of treated celiac disease. This study examined the endoscopic and histopathologic appearance of the duodenum of patients with celiac disease whose diet was gluten-free. METHODS: A cohort of 39 adult patients (mean age 52 years, range 20-74 years) with biopsy-proven celiac disease was retrospectively reviewed. All had responded clinically to a gluten-free diet that they had maintained for a mean of 8.5 years (range 1-45 years). The endoscopic and histopathologic appearances of the duodenal mucosa were reviewed. Blinded review of the diagnostic (initial) and post-treatment biopsy specimens was also performed to assess response of individual patients to the diet. RESULTS: The endoscopic appearance was normal in 23%, reduced duodenal folds were present in 46%, scalloping of folds in 33%, mucosal fissures in 44%, and nodularity in 33%. There was more than 1 abnormality present in 46%. Histology was normal in only 21%. The remainder had villous atrophy (69% partial, 10% total). Paired (diagnostic and follow-up) biopsy specimens were reviewed blindly for 12 patients. The mean (SD) intraepithelial lymphocyte count fell from 61 (22) to 38 (17) (normal <30 per 100 epithelial cells) and the crypt-to-villous ratio improved although it did not normalize. CONCLUSIONS: Despite a good clinical response, abnormal endoscopic and histopathologic appearances persist in the majority of patients with celiac disease treated with a gluten-free diet. PMID- 12556783 TI - A comparative study of postendoscopic sphincterotomy complications with various types of electrosurgical current in patients with choledocholithiasis. AB - BACKGROUND: Whether the type of electrosurgical current used for endoscopic sphincterotomy influences the frequency of postsphincterotomy complications is unknown. METHODS: One hundred eighty-six patients with choledocholithiasis were prospectively randomized to undergo endoscopic sphincterotomy with pure cutting current (n = 62, Group A), blended current (n = 62, Group B), or pure cutting initially followed by blended current (n = 62, Group C). Serum concentrations of amylase and lipase were evaluated in all patients 12 and 24 hours after sphincterotomy. Clinical pancreatitis was classified as mild, moderate, or severe. Postsphincterotomy bleeding was defined as a decrease in hematocrit of greater than 5%. RESULTS: Serum concentrations of amylase and lipase were greater in Groups B and C at 12 and 24 hours after the procedure, as compared with Group A. Clinical mild pancreatitis occurred in 2 patients in Group A (3.2%), 8 in Group B (12.9%), and in 8 in Group C (12.9%). The differences were statistically significant for Group A compared with either Group B or Group C (p = 0.048). Postsphincterotomy bleeding occurred in 3 patients (1.6%), one in each group. CONCLUSION: The use of pure cutting electrosurgical current during endoscopic sphincterotomy in patients with choledocholithiasis is associated with a lesser degree of pancreatic enzyme elevation and lower frequency of pancreatitis, whereas bleeding is not increased compared with blended current. Changing from pure cutting to blended current after the first 3 to 5 mm of the incision is associated with an increased rate of complications compared to the use of pure cutting current for the entire sphincterotomy. PMID- 12556784 TI - Unsedated transnasal EGD in daily practice: results with 1100 consecutive patients. AB - BACKGROUND: EGD can be performed transnasally in adults by using small-diameter endoscopes. A large prospective study was conducted to evaluate the feasibility and tolerance of diagnostic transnasal EGD in daily practice. METHODS: Unsedated transnasal EGD was attempted in 1100 consecutive patients, in 3 different institutions, by using a 5.9-mm or a 5.3-mm diameter endoscope. The operator determined whether the procedure was successful or unsuccessful, the reason for failures, and any side effects. The influence of gender, age, endoscope diameter, and type of topical anesthesia on the success or failure of the procedure was evaluated. Patients who previously had undergone peroral EGD were queried as to which procedure they preferred. RESULTS: Transnasal EGD was feasible in 93.9% of the patients. The causes of failure were as follows: unsuccessful transnasal insertion (62.7%), patient refusal (19.4%), and nasal pain (17.9%). Female gender, young age (< or =35 years), and larger-endoscope diameter were significant predictive factors for procedure failure. Side effects included the following: epistaxis (2.3%), nasal pain (1.6%), and vaso-vagal reaction (0.3%). A majority (91%) of the patients who previously had undergone unsedated peroral EGD with a standard 9.8-mm diameter endoscope preferred transnasal EGD with a small diameter endoscope. CONCLUSIONS: Transnasal EGD is feasible in daily endoscopic practice and is preferred by patients. Side effects are rare. PMID- 12556785 TI - Efficacy of peroral pancreatoscopy in the diagnosis of pancreatic diseases. AB - BACKGROUND: The aim of this study was to evaluate the usefulness of peroral pancreatoscopy in the diagnosis of pancreatic diseases. METHODS: Both 3.7-mm (thin) and 0.8-mm (ultra-thin) diameter fiberoptic pancreatoscopes were used in 115 cases (pancreatic cancer, 35; benign ductal stenosis, 20; intraductal papillary-mucinous tumor, 60). RESULTS: Observation rates for pancreatic cancer, benign ductal stenosis, and intraductal papillary-mucinous tumor were, respectively, 63%, 80%, and 95%. Tumor vessels and papillary tumor were observed when pancreatic cancer was smaller than 2 cm but not when the tumor was larger than 2 cm. Stenosis without significant mucosal changes was observed in 62% of cases of benign ductal stenosis. Coarse mucosa and friability were observed more frequently in association with pancreatic cancer than benign ductal stenosis. Granular mucosa or papillary tumor could be observed in 74% of cases of intraductal papillary-mucinous tumor. Papillary tumor was observed with increasing frequency in cases of intraductal papillary-mucinous tumor as the degree of malignancy increased. CONCLUSIONS: Peroral pancreatoscopy with an ultra thin fiberscope is useful in the diagnosis of minute pancreatic lesions. Peroral pancreatoscopy with a thin fiberscope can provide a definitive diagnosis of intraductal papillary-mucinous tumor including the degree of malignancy. PMID- 12556786 TI - ERCP in the diagnosis and management of complications after surgery for hepatic echinococcosis. AB - BACKGROUND: Surgery for liver hydatidosis can result in serious morbidity and mortality. The role and efficacy of ERCP in the management of these complications was reviewed. METHODS: Retrospective analysis of 79 patients with liver hydatidosis who underwent ERCP for postoperative complications. RESULTS: All patients with biliary fistulas (n = 50) were managed by endoscopic sphincterotomy, although 9 (18%) also required a biliary endoprosthesis. Surgical reintervention was avoided in 46 (92%) patients with subsequent surgery required in only in 4 (8%). The mean time to fistula closure was 13.3 days. Patients with daughter cysts in the bile ducts (n = 7) were also treated by endoscopic sphincterotomy and evacuation of obstructing cyst material. Seven patients with elevated liver enzymes or biliopancreatic symptoms but normal ERCP findings also underwent endoscopic sphincterotomy with normalization of biochemical tests. CONCLUSIONS: ERCP is valuable for detecting and treating postoperative biliary complications after surgery for hepatic echinococcal disease. In the majority of patients, endoscopic sphincterotomy allows healing of postoperative external biliary fistulas, and should be performed as early as possible. In some cases, a biliary prosthesis may be required. Endoscopic sphincterotomy also enables clearing of the bile ducts of hydatid remnants. PMID- 12556787 TI - Validation of a computer-based colonoscopy simulator. AB - BACKGROUND: The computer-based colonoscopy simulator is intended to provide a realistic colonoscopic experience and feedback to operators regarding procedure skills. Advocates hope that computer-based colonoscopy simulators will enhance the mastery of colonoscopy by trainees. Before this hypothesis can be tested, the claims made for a simulator must be validated. The aims of this study were to answer the following: Does a computer-based colonoscopy simulator provide a "realistic" experience? Do computer-based colonoscopy simulators' performance parameters differentiate varying levels of experience? METHODS: Ten staff gastroenterologists, 6 gastroenterology fellows, and 6 residents each performed 2 computer-based colonoscopy simulator colonoscopies and performance parameters were recorded. Staff colonoscopists then completed a 6-item survey grading the "realism" of the simulation and procedure difficulty. Survey responses and performance scores were compared with the Wilcoxon rank-sum test. RESULTS: Faculty found the computer-based colonoscopy simulator experience to be realistic despite the "cases" being markedly easier than actual colonoscopy. The computer based colonoscopy simulator distinguishes subjects according to endoscopic experience with 3 of its measured parameters (total procedure time, insertion time, time in "red-out"). No significant difference in the ability to distinguish among user types was found for the other 10 computer-based colonoscopy simulator measurements for which data were analyzable. CONCLUSION: The computer-based colonoscopy simulator provides a favorable degree of virtual realism with regard to visual simulation and colonoscope mechanics, although the "cases" were regarded as considerably easier than actual colonoscopy. The computer-based colonoscopy simulator has only limited capability for distinguishing varying levels of competence at actual colonoscopy. These findings suggest that a study to determine the role of computer-based colonoscopy simulators in the curriculum of trainees is warranted. PMID- 12556788 TI - Botulinum toxin for spastic GI disorders: a systematic review. PMID- 12556789 TI - Pelvic actinomycosis. PMID- 12556790 TI - Gastric corrosive injury. PMID- 12556791 TI - Granulocytic sarcoma of esophagus. PMID- 12556793 TI - Choledochocele and common bile duct stones. PMID- 12556792 TI - Toothpick penetration of stomach. PMID- 12556794 TI - Pneumatosis cystoides intestinalis in Crohn's disease. PMID- 12556795 TI - Newly designed soft prelooped cap for endoscopic mucosal resection of gastric lesions. AB - BACKGROUND: Endoscopic mucosal resection with a cap-fitted panendoscope is a useful, effective, and safe technique. The effectiveness and safety of a newly designed larger but softer cap was compared in this study with the conventional hard cap. The soft cap has a significantly larger diameter (18 mm) compared with that of the hard cap (16.5 mm). METHODS: Eighty-three patients who underwent mucosal resection of gastric lesions were included in analysis. The diameter, resection specimen depth, and the rate of en bloc resection were compared. The operability of the endoscope with the cap attached, patient tolerance, and safety of the procedure were analyzed. RESULTS: Mean diameter (+/- SEM) of specimens resected with the soft cap was larger: 22.1 (+/- 0.7) versus 15.8 (+/- 0.3) mm (p < 0.001). The specimen was also thicker: 1.54 (+/- 0.10) versus 1.08 (+/- 0.11) mm (p < 0.001). Use of the soft cap led to higher rate of en bloc resection: 66.7% versus 43.2% (p < 0.05). The operability of the endoscope with the larger, softer cap attached was similar to that when the hard cap was used. Both caps were equally safe. CONCLUSION: The soft cap is safe and useful for mucosal resection of larger gastric lesions. Its use increases the rate of en bloc resection. PMID- 12556796 TI - EUS-cholangio-drainage of the bile duct: report of 4 cases. AB - BACKGROUND: Endoscopic retrograde cholangiography with stent insertion is an established procedure for palliation of patients with malignant pancreaticobiliary strictures. In some patients, however, placement of a stent by means of a duodenoscope is not possible. Percutaneous transhepatic biliary drainage is an alternative method that has a complication rate of up to 15%. Four cases of successful EUS-guided-cholangio-drainage are presented in which the major papilla could not be cannulated at endoscopic retrograde cholangiography. METHODS: For puncture of the intrahepatic or extrahepatic bile duct, a modification of the one-step technique for the drainage of pancreatic pseudocysts was used. RESULTS: Stent insertion was successful in 3 of the 4 patients. In these 3 patients cholestasis resolved promptly. CONCLUSIONS: EUS-guided cholangio drainage is a potential alternative to percutaneous transhepatic biliary drainage and endoscopic retrograde cholangiography in the therapy of malignant pancreaticobiliary strictures, especially in patients who have undergone gastrectomy or partial gastrectomy with Billroth II reconstruction. PMID- 12556797 TI - EUS-guided FNA for the diagnosis of gallbladder masses. AB - BACKGROUND: Gallbladder masses can be identified endosongraphically, but FNA for cytologic diagnosis is not routine. This is a review of our experience with EUS guided FNA of gallbladder masses. METHODS: Records of patients undergoing EUS were reviewed to identify cases in which FNA of the gallbladder was performed. Reports of EUS procedures, EUS images, cytology results, and clinical records were reviewed. OBSERVATIONS: Six cases were identified. The final diagnosis was gallbladder carcinoma in 5 and xanthogranulomatous cholecystitis in one. In each case, EUS revealed a hypoechoic mass within the gallbladder wall or gallbladder lumen. Gallbladder wall calcification was observed in 3 of the 5 cases of carcinoma. FNA yielded a specimen that was positive (n = 3) or raised a suspicion (n = 1) for adenocarcinoma in 4 of the 5 proven malignancies. FNA of regional lymph nodes demonstrated metastatic adenocarcinoma in 2 cases. FNA was negative for malignancy in the case of xanthogranulomatous cholecystitis and one case of proven carcinoma. There were no complications. CONCLUSIONS: EUS-guided FNA of gallbladder masses is safe and can provide a definitive diagnosis of malignancy. Gallbladder carcinoma appears endosonographically as a hypoechoic mass and may be associated with focal wall calcifications. PMID- 12556798 TI - Intramural cecal hematoma: an unusual complication of colonoscopy. PMID- 12556799 TI - Ischemic colitis, systemic lupus erythematosus, and the lupus anticoagulant: case report and review. PMID- 12556800 TI - Hemobilia caused by a mycotic aneurysm of the hepatic artery treated by enbucrilate injection during ERCP. PMID- 12556801 TI - Endoscopic mucosal resection of adenocarcinoma arising in ectopic gastric mucosa in the cervical esophagus: case report. PMID- 12556802 TI - Photodynamic therapy with polypectomy for rectal cancer. PMID- 12556803 TI - Acute portal vein thrombosis after EUS-guided FNA of pancreatic cancer: case report. PMID- 12556804 TI - Bowel perforation with the variable stiffness colonoscope. PMID- 12556805 TI - Small bowel obstruction caused by bowel wall hematoma after PEG. PMID- 12556806 TI - Dysphagia caused by transluminal erosion of a titanium rod into the esophagus. PMID- 12556807 TI - Endoscopic correction of a buried tracheo-esophageal prosthesis. PMID- 12556808 TI - "Spontaneous" passage of a pancreatic duct stone. PMID- 12556809 TI - Ring around the cardia: a watermelon stomach variant. PMID- 12556810 TI - Endoscopic balloon dilation for extraction of bile duct stones: the devil is in the details. PMID- 12556811 TI - New horizons. PMID- 12556812 TI - Defined nomenclature of pancreatic fluid collections. PMID- 12556814 TI - Peptic ulcer, Helicobacter pylori, and reflux symptoms. PMID- 12556816 TI - Rectal digitation and solitary rectal ulcer. PMID- 12556817 TI - Centuries of history...centuries of caring. PMID- 12556818 TI - A call to arms, a call for caution. PMID- 12556821 TI - Focused abdominal sonogram for trauma to identify blood within the abdomen. PMID- 12556822 TI - A 67-year-old man with increasing severe lower back pain since the night before. PMID- 12556823 TI - Improving medication safety and patient care in the emergency department. AB - INTRODUCTION: Medication errors are well documented in medical literature and the lay press. Through participation in a nationwide institute for healthcare improvement initiative, our emergency department performance improvement group focused on human and system factors that contributed to potential medication errors. METHODS: A survey conducted of ED staff examined barriers to reporting medication errors and potential "near misses." members of the emergency department performance improvement group examined contents of the ed Pyxis machines, assessing medications that physically resembled one another, similar sounding medications located in close proximity, and medications available in differing doses. RESULTS: Fifty-eight members participated in a 4-question survey. Half reported they would be likely to self-report a "near miss" if the patient was not harmed. About half would report the medication error of a colleague under certain circumstances. Fifty-one percent believed there would be repercussions for reporting medication error, but most believed they would receive support from supervisors for addressing other safety problems. Nearly one quarter of the 278 medications identified in the Pyxis survey were similar in appearance or name or existed in multidose formulations. DISCUSSION: Measures to decrease the potential of medication errors include: (1) a workplace environment that promotes reporting of medication errors or "close calls" by staff, with counseling events utilized as learning opportunities versus punitive incidents; (2) increased frequency of medication safety in-service sessions; and (3) periodic monitoring of Pyxis machine inventories to survey contents for optimum patient safety. PMID- 12556824 TI - Serendipitous findings from an Illinois ED nursing educational survey reflect a crisis in emergency nursing. AB - INTRODUCTION: A survey conducted by the Research Council Of Illinois ENA was intended to assess educational needs, but it revealed some very important serendipitous findings. This survey gave a voice to the frustrations of emergency nurses. METHOD: A 31-question survey was distributed to 900 Illinois ENA members requesting information about their educational needs. The last question, which formed the basis of this report, asked respondents to list the 3 biggest challenges they face on a daily basis. RESULTS: In addition to the information regarding educational offerings, the survey revealed that ED nurses were overburdened and frustrated in their jobs. Specifically, the nurses identified 7 areas of greatest concern: (1) insufficient and inexperienced staff; (2) increased responsibilities; (3) lack of administrative support; (4) lack of rewards or incentives to stay; (5) low morale among staff; (6) difficulty balancing work and family; and (7) increasing violence in the emergency department. DISCUSSION: The first step toward addressing a growing crisis in emergency nursing is to listen to the voices of emergency nurses themselves and let their frustrations be heard. The spontaneous comments of emergency nurses point to serious, pervasive, systemic problems in emergency nursing. Such problems will not be solved by "quick fixes" such as sign on bonuses, but rather by listening carefully to the concerns of experienced nurses. PMID- 12556825 TI - Work stress and posttraumatic stress disorder in ED nurses/personnel. AB - INTRODUCTION: Work-related stress in the emergency department previously has been linked to depression and burnout; however, these findings have not been extended to the development of anxiety disorders, such as posttraumatic stress disorder (PTSD). Three sets of factors have been shown to contribute to stress in ED personnel: organizational characteristics, patient care, and the interpersonal environment. The current study addressed whether an association exists between sources of workplace stress and PTSD symptoms. METHOD: Respondents were 51 ED personnel from a hospital in a large Canadian urban center. The majority of respondents were emergency nurses. Respondents completed questionnaires measuring PTSD and sources of work stress and answered a series of questions regarding work related responses to stress or trauma. RESULTS: Interpersonal conflict was significantly associated with PTSD symptoms. The majority of respondents (67%) believed they had received inadequate support from hospital administrators following the traumatic incident and 20% considered changing jobs as a result of the trauma. Only 18% attended critical incident stress debriefing and none sought outside help for their distress. DISCUSSION: These findings underscore the need for hospital administrations to be aware of the extent of workplace stress and PTSD symptoms in their employees. Improving the interpersonal climate in the workplace may be useful in ameliorating PTSD symptoms. PMID- 12556826 TI - Unraveling the mysteries of mechanical ventilation: a helpful step-by-step guide. PMID- 12556827 TI - An emergency nurse's work with refugees in Africa, The Balkans, and Afghanistan. PMID- 12556829 TI - Electronic tracking board reduces ED patient length of stay at Indiana Hospital. PMID- 12556831 TI - "Code Silver:" an EMTALA compliant response to ill or injured nonpatients on hospital property. PMID- 12556833 TI - Child car seat checkpoints: one urban community's experience. PMID- 12556834 TI - Encounter at triage results in legal liability. PMID- 12556837 TI - A combined "crossover" course for TNCC and ENPC: an innovative approach to preparing more TNCC and ENPC instructors. PMID- 12556838 TI - A 3-month-old with blood in the stool: a case scenario. PMID- 12556839 TI - GHB poisoning: three recent cases reflect the continuing danger. PMID- 12556840 TI - UCSF's new center for tobacco control research and education finds valuable lessons in the tobacco industry's internal documents. PMID- 12556841 TI - Professional nursing council empowers staff nurses to solve clinical problems and increases RN retention. PMID- 12556842 TI - Snakebite poisoning: treatment myths and facts. PMID- 12556843 TI - Resuscitating trauma patients in the intensive care unit. PMID- 12556844 TI - Lessons learned: On watching for zebras. PMID- 12556846 TI - All the President's yes-men? PMID- 12556847 TI - Too quiet on the Eastern front. PMID- 12556848 TI - Gates ploughs millions into plan for assault on killer diseases. PMID- 12556849 TI - UK cell biologist takes pole position at Rockefeller. PMID- 12556850 TI - Transgenic crop trial's gene flow turns weeds into wimps. PMID- 12556851 TI - Head of Spanish lab network quits over lack of funds. PMID- 12556852 TI - TB medic wins global health post. PMID- 12556853 TI - US army survey targets Gulf War syndrome. PMID- 12556854 TI - Taiwanese institute causes upset by degrees. PMID- 12556855 TI - Scientists call for Canada to boost polar-shelf funding. PMID- 12556856 TI - Europe seeks single defence research agency. PMID- 12556857 TI - Italians riled by science reforms. PMID- 12556859 TI - Oceanography: Robots in the deep. PMID- 12556860 TI - Polish science: Poles apart, or together with Europe? PMID- 12556861 TI - Venezuelan researchers call for international help. PMID- 12556862 TI - No reduction in risk of a massive asteroid impact. PMID- 12556863 TI - Was visionary Seaborg from far-sighted stock? PMID- 12556864 TI - Is a picture worth 1,000 words? PMID- 12556870 TI - Geometric frustration: Magic moments. PMID- 12556871 TI - Particle physics: Now you see them, now you don't. PMID- 12556872 TI - DNA repair: Damage alert. PMID- 12556873 TI - Astronomy: Distant planet is the hottest yet. PMID- 12556874 TI - Biodiversity: The threat of small households. PMID- 12556875 TI - Condensed-matter physics: Tunable colloidal crystals. PMID- 12556876 TI - Behavioural genetics: Family matters. PMID- 12556879 TI - Neurophysiology: Sensing temperature without ion channels. PMID- 12556880 TI - Nanoelectromechanical systems: Nanodevice motion at microwave frequencies. PMID- 12556881 TI - Genomic function: Rate of evolution and gene dispensability. PMID- 12556883 TI - Cosmology: Do black holes constrain varying constants? PMID- 12556884 TI - DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation. AB - The ATM protein kinase, mutations of which are associated with the human disease ataxia-telangiectasia, mediates responses to ionizing radiation in mammalian cells. Here we show that ATM is held inactive in unirradiated cells as a dimer or higher-order multimer, with the kinase domain bound to a region surrounding serine 1981 that is contained within the previously described 'FAT' domain. Cellular irradiation induces rapid intermolecular autophosphorylation of serine 1981 that causes dimer dissociation and initiates cellular ATM kinase activity. Most ATM molecules in the cell are rapidly phosphorylated on this site after doses of radiation as low as 0.5 Gy, and binding of a phosphospecific antibody is detectable after the introduction of only a few DNA double-strand breaks in the cell. Activation of the ATM kinase seems to be an initiating event in cellular responses to irradiation, and our data indicate that ATM activation is not dependent on direct binding to DNA strand breaks, but may result from changes in the structure of chromatin. PMID- 12556885 TI - An extrasolar planet that transits the disk of its parent star. AB - Planets orbiting other stars could in principle be found through the periodic dimming of starlight as a planet moves across--or 'transits'--the line of sight between the observer and the star. Depending on the size of the planet relative to the star, the dimming could reach a few per cent of the apparent brightness of the star. Despite many searches, no transiting planet has been discovered in this way; the one known transiting planet--HD209458b--was first discovered using precise measurements of the parent star's radial velocity and only subsequently detected photometrically. Here we report radial velocity measurements of the star OGLE-TR-56, which was previously found to exhibit a 1.2-day transit-like light curve in a survey looking for gravitational microlensing events. The velocity changes that we detect correlate with the light curve, from which we conclude that they are probably induced by an object of around 0.9 Jupiter masses in an orbit only 0.023 au from its star. We estimate the planetary radius to be around 1.3 Jupiter radii and its density to be about 0.5 g x cm(-3). This object is hotter than any known planet (approximately 1,900 K), but is still stable against long-term evaporation or tidal disruption. PMID- 12556886 TI - Long-distance teleportation of qubits at telecommunication wavelengths. AB - Matter and energy cannot be teleported (that is, transferred from one place to another without passing through intermediate locations). However, teleportation of quantum states (the ultimate structure of objects) is possible: only the structure is teleported--the matter stays at the source side and must be already present at the final location. Several table-top experiments have used qubits (two-dimensional quantum systems) or continuous variables to demonstrate the principle over short distances. Here we report a long-distance experimental demonstration of probabilistic quantum teleportation. Qubits carried by photons of 1.3 micro m wavelength are teleported onto photons of 1.55 micro m wavelength from one laboratory to another, separated by 55 m but connected by 2 km of standard telecommunications fibre. The first (and, with foreseeable technologies, the only) application of quantum teleportation is in quantum communication, where it could help to extend quantum cryptography to larger distances. PMID- 12556887 TI - A colloidal model system with an interaction tunable from hard sphere to soft and dipolar. AB - Monodisperse colloidal suspensions of micrometre-sized spheres are playing an increasingly important role as model systems to study, in real space, a variety of phenomena in condensed matter physics--such as glass transitions and crystal nucleation. But to date, no quantitative real-space studies have been performed on crystal melting, or have investigated systems with long-range repulsive potentials. Here we demonstrate a charge- and sterically stabilized colloidal suspension--poly(methyl methacrylate) spheres in a mixture of cycloheptyl (or cyclohexyl) bromide and decalin--where both the repulsive range and the anisotropy of the interparticle interaction potential can be controlled. This combination of two independent tuning parameters gives rise to a rich phase behaviour, with several unusual colloidal (liquid) crystalline phases, which we explore in real space by confocal microscopy. The softness of the interaction is tuned in this colloidal suspension by varying the solvent salt concentration; the anisotropic (dipolar) contribution to the interaction potential can be independently controlled with an external electric field ranging from a small perturbation to the point where it completely determines the phase behaviour. We also demonstrate that the electric field can be used as a pseudo-thermodynamic temperature switch to enable real-space studies of melting transitions. We expect studies of this colloidal model system to contribute to our understanding of, for example, electro- and magneto-rheological fluids. PMID- 12556888 TI - High-temperature superconductor bulk magnets that can trap magnetic fields of over 17 tesla at 29 K. AB - Large-grain high-temperature superconductors of the form RE-Ba-Cu-O (where RE is a rare-earth element) can trap magnetic fields of several tesla at low temperatures, and so can be used for permanent magnet applications. The magnitude of the trapped field is proportional to the critical current density and the volume of the superconductor. Various potential engineering applications for such magnets have emerged, and some have already been commercialized. However, the range of applications is limited by poor mechanical stability and low thermal conductivity of the bulk superconductors; RE-Ba-Cu-O magnets have been found to fracture during high-field activation, owing to magnetic pressure. Here we present a post-fabrication treatment that improves the mechanical properties as well as thermal conductivity of a bulk Y-Ba-Cu-O magnet, thereby increasing its field-trapping capacity. First, resin impregnation and wrapping the materials in carbon fibre improves the mechanical properties. Second, a small hole drilled into the centre of the magnet allows impregnation of Bi-Pb-Sn-Cd alloy into the superconductor and inclusion of an aluminium wire support, which results in a significant enhancement of thermal stability and internal mechanical strength. As a result, 17.24 T could be trapped, without fracturing, in a bulk Y-Ba-Cu-O sample of 2.65 cm diameter at 29 K. PMID- 12556889 TI - The effects of surfactants on spilling breaking waves. AB - Breaking waves markedly increase the rates of air-sea transfer of momentum, energy and mass. In light to moderate wind conditions, spilling breakers with short wavelengths are observed frequently. Theory and laboratory experiments have shown that, as these waves approach breaking in clean water, a ripple pattern that is dominated by surface tension forms at the crest. Under laboratory conditions and in theory, the transition to turbulent flow is triggered by flow separation under the ripples, typically without leading to overturning of the free surface. Water surfaces in nature, however, are typically contaminated by surfactant films that alter the surface tension and produce surface elasticity and viscosity. Here we present the results of laboratory experiments in which spilling breaking waves were generated mechanically in water with a range of surfactant concentrations. We find significant changes in the breaking process owing to surfactants. At the highest concentration of surfactants, a small plunging jet issues from the front face of the wave at a point below the wave crest and entraps a pocket of air on impact with the front face of the wave. The bubbles and turbulence created during this process are likely to increase air-sea transfer. PMID- 12556890 TI - Damage to the shallow Landers fault from the nearby Hector Mine earthquake. AB - Crustal faults have long been identified as sites where localized sliding motion occurs during earthquakes, which allows for the relative motion between adjacent crustal blocks. Although there is a growing awareness that we must understand the evolution of fault systems on many timescales to relate present-day crustal stresses and fault motions to geological structures formed in the past, fault zone damage and healing have been documented quantitatively in only a few cases. We have been monitoring the healing of damage on the shallow Johnson Valley fault after its rupture in the 1992 magnitude-7.3 Landers earthquake, and here we report that this healing was interrupted in 1999 by the magnitude-7.1 Hector Mine earthquake rupture, which occurred 20-30 km away. The Hector Mine earthquake both strongly shook and permanently strained the Johnson Valley fault, adding damage discernible as a temporary reversal of the healing process. The fault has since resumed the trend of strength recovery that it showed after the Landers earthquake. These observations lead us to speculate that fault damage caused by strong seismic waves may help to explain earthquake clustering and seismicity triggering by shaking, and may be involved in friction reduction during faulting. PMID- 12556891 TI - Head and backbone of the Early Cambrian vertebrate Haikouichthys. AB - Agnathan fish hold a key position in vertebrate evolution, especially regarding the origin of the head and neural-crest-derived tissue. In contrast to amphioxus, lampreys and other vertebrates possess a complex brain and placodes that contribute to well-developed eyes, as well as auditory and olfactory systems. These sensory sytems were arguably a trigger to subsequent vertebrate diversifications. However, although they are known from skeletal impressions in younger Palaeozoic agnathans, information about the earliest records of these systems has been largely wanting. Here we report numerous specimens of the Lower Cambrian vertebrate Haikouichthys ercaicunensis, until now only known from the holotype. Haikouichthys shows significant differences from other fossil agnathans: key features include a small lobate extension to the head, with eyes and possible nasal sacs, as well as what may be otic capsules. A notochord with separate vertebral elements is also identifiable. Phylogenetic analysis indicates that this fish lies within the stem-group craniates. Although Haikouichthys somewhat resembles the ammocoete larva of modern lampreys, this is because of shared general craniate characters; adult lampreys and hagfishes (the cyclostomes if monophyletic) are probably derived in many respects. PMID- 12556892 TI - The genetic basis of family conflict resolution in mice. AB - Asymmetries in the costs and benefits of parental investment for mothers, fathers and offspring result in family conflict over the production and provisioning of young. In species where females provide most resources before and after birth, the resolution of this conflict may be influenced by genes expressed in mothers and by maternally and paternally inherited genes expressed in offspring. Here we disentangle these effects by means of reciprocal mating and cross-fostering of litters between two strains of mice that differ with respect to the typical resolution of family conflict. We find that differences in litter size between these two strains are determined by paternal genotype, whereas differences in provisioning are under maternal control, showing that there is antagonistic coadaptation of maternal and paternal effects on distinct life-history traits. Maternal provisioning is also influenced by the type of foster offspring. Contradictory to theoretical expectations, however, we find no evidence for a negative correlation across strains between maternal provisioning and offspring demand. Instead, we show that there is positive coadaptation such that offspring obtain more resources from foster mothers of the same strain as their natural mother, irrespective of their father's strain. PMID- 12556893 TI - A spatially organized representation of colour in macaque cortical area V2. AB - Neurons responding selectively to different colours have been found in various cortical areas in macaque monkeys; however, little is known about whether and how the representation of colour is spatially organized in any cortical area. Cortical area V2 contains modules that respond preferentially to chromatic modulation, which are located in thin cytochrome oxidase stripes. Here we show that within and beyond these modules, gratings of different colours produce activations that peak at different locations. Optical recording of intrinsic signals revealed that the peak regions of the responses to different colours were spatially organized in the same order as colour stimuli are arranged in the DIN (German standard colour chart) colour system. Nearby regions represented colours of a similar hue. We found that the set of colour-specific regions formed 0.07 0.32-mm-wide and approximately 1.3-mm long bands that varied in shape from linear to nearly circular. Our finding suggests that thin stripes in V2 contain functional maps where the colour of a stimulus is represented by the location of its response activation peak. PMID- 12556894 TI - Inhibitory feedback required for network oscillatory responses to communication but not prey stimuli. AB - Stimulus-induced oscillations occur in visual, olfactory and somatosensory systems. Several experimental and theoretical studies have shown how such oscillations can be generated by inhibitory connections between neurons. But the effects of realistic spatiotemporal sensory input on oscillatory network dynamics and the overall functional roles of such oscillations in sensory processing are poorly understood. Weakly electric fish must detect electric field modulations produced by both prey (spatially localized) and communication (spatially diffuse) signals. Here we show, through in vivo recordings, that sensory pyramidal neurons in these animals produce an oscillatory response to communication-like stimuli, but not to prey-like stimuli. On the basis of well-characterized circuitry, we construct a network model of pyramidal neurons that predicts that diffuse delayed inhibitory feedback is required to achieve oscillatory behaviour only in response to communication-like stimuli. This prediction is experimentally verified by reversible blockade of feedback inhibition that removes oscillatory behaviour in the presence of communication-like stimuli. Our results show that a sensory system can use inhibitory feedback as a mechanism to 'toggle' between oscillatory and non-oscillatory firing states, each associated with a naturalistic stimulus. PMID- 12556896 TI - Visions of Europe's future. PMID- 12556898 TI - Genetics, genes, genomics and g. PMID- 12556901 TI - Role of the cholinergic muscarinic 2 receptor (CHRM2) gene in cognition. PMID- 12556902 TI - No association of alcohol dependence with a NMDA-receptor 2B gene variant. PMID- 12556903 TI - Clozapine-induced reduction of glutamate transport in the frontal cortex is not mediated by GLAST and EAAC1. PMID- 12556905 TI - Neonatal perturbation of neurotrophic signaling results in abnormal sensorimotor gating and social interaction in adults: implication for epidermal growth factor in cognitive development. AB - Epidermal growth factor (EGF) and its structurally related proteins are implicated in the developmental regulation of various brain neurons, including midbrain dopaminergic neurons. There are EGF and EGF receptor abnormalities in both brain tissues and blood from schizophrenic patients. We administered EGF to neonatal rats to transiently perturb endogenous EGF receptor signaling and evaluated the neurobehavioral consequences. EGF-treatment-induced transient impairment in tyrosine hydroxylase expression. The animals grew normally, exhibited normal weight increase, glial growth, and gross brain structures, and later lost the tyrosine hydroxylase abnormality. During and after development, however, the rats began to display various behavioral abnormalities. Abnormal sensorimotor gating was apparent, as measured by deficits in prepulse inhibition of acoustic startle. Motor activity and social interaction scores of the EGF treated animals were also impaired in adult rats, though not in earlier developmental stages. In parallel, there was a significant abnormality in dopamine metabolism in the brain stem of the adult animals. Gross learning ability appeared to be normal as measured by active avoidance. These behavioral alterations, which are often present in schizophrenic models, were ameliorated by subchronic treatment with clozapine. Although the molecular and/or physiologic background(s) of these behavioral abnormalities await further investigation, the results of the present experiment indicate that abnormal EGF receptor stimulation given during limited neonatal stages can result in severe and persistent cognitive/behavioral dysfunctions, which appear only in adulthood. PMID- 12556904 TI - Cathepsin D exon 2 polymorphism associated with general intelligence in a healthy older population. AB - General intelligence is a heritable trait that is a risk factor for both the onset of dementia and the rate of cognitive decline in community-dwelling older persons. Previous studies screening for quantitative trait loci (QTLs) that influence general intelligence in healthy individuals have identified four loci, two of which are located within the genes insulin-like growth factor 2 receptor (IGF2R) and the Msx1 homeobox. Here, we report the finding of another QTL associated with general intelligence that is located within exon 2 of the cathepsin D (CTSD) gene. A group of 767 healthy adults with a follow-up period of over 15 years have been analyzed for cross-sectional and longitudinal trends in cognitive change using the Heim intelligence test score (AH4-1). We observed a significant association (P = 0.01) between a functional C > T (Ala > Val) transition within exon 2 of the CTSD gene that increases the secretion of pro CTSD from the cell, and the AH4-1 score at initial testing on entry to the longitudinal study. Interestingly, CTSD is transported by IGF2R from the trans Golgi network to the lysosome. PMID- 12556906 TI - Lack of thyroid hormone receptor alpha1 is associated with selective alterations in behavior and hippocampal circuits. AB - Brain development and function are dependent on thyroid hormone (T3), which acts through nuclear hormone receptors. T3 receptors (TRs) are transcription factors that activate or suppress target gene expression in a hormone-dependent or independent fashion. Two distinct genes, TRalpha and TRbeta, encode several receptor isoforms with specific functions defined in many tissues but not in the brain. Mutations in the TRbeta gene cause the syndrome of peripheral resistance to thyroid hormone; however, no alterations of the TRalpha gene have been described in humans. Here we demonstrate that mice lacking the TRalpha1 isoform display behavioral abnormalities of hippocampal origin, as shown by the open field and fear conditioning tests. In the open field test mutant mice revealed less exploratory behavior than wild-type mice. In the contextual fear conditioning test mutant mice showed a significantly higher freezing response than wild-type controls when tested 1 week after training. These findings correlated with fewer GABAergic terminals on the CA1 pyramidal neurons in the mutant mice. Our results indicate that TRalpha1 is involved in the regulation of hippocampal structure and function, and raise the possibility that deletions or mutations of this receptor isoform may lead to behavioral changes or even psychiatric syndromes in humans. PMID- 12556907 TI - Cloning of murine TReP-132, a novel transcription factor expressed in brain regions involved in behavioral and psychiatric disorders. AB - In recent studies that addressed the transcriptional control of steroid synthesis, a transcriptional regulating protein of 132 kDa (TReP-132) was cloned and demonstrated to regulate expression of the human P450 side chain cleavage (P450scc) gene. In the present study, we describe the cloning and characterization of the mouse orthologue of the human factor, mouse transcriptional regulating protein (mTReP-132). mTReP-132 encodes a 1216-residue protein that is 85.5% homologous to the human protein. Both factors contain characteristic motifs, namely glutamine-, proline- and acidic-rich regions. The primary structure also exhibits two zinc fingers of the C(2)H(2) subtype, suggesting that this protein has the ability to act as a DNA binding transcription factor. mTReP-132 may also be a co-regulator of nuclear receptors because of two nuclear box motifs in this protein. Northern blot analysis demonstrated the expression of two transcripts of 4.4 and 7.5 kb in several tissues, but expression was clearly highest in the brain, thymus and testis of mice. In the brain, the hybridization signal was quite localized and strong in the basal ganglia, hippocampus, piriform cortex, cerebral cortex, ventromedial nucleus of the hypothalamus, and the dorsal and superior central nuclei of the raphe. Although classical steroidogenesis pathways have yet to be firmly established in the brain, expression of both mTReP-132 and P450scc provides anatomical evidence that mTReP-132 may regulate this key steroidogenic enzyme within specific regions involved in behavioral and psychiatric disorders. Moreover, the presence of both mTReP-132 and steroidogenic factor 1 (SF-1) transcripts in the ventromedial nucleus of the hypothalamus suggests a role for mTReP-132 in brain development and function. The molecular cloning and the highly specific expression of mTReP-132 across the brain further consolidate the hypothesis that this tissue is able to synthesize de novo steroids in a region specific manner. PMID- 12556908 TI - Blockade of mesolimbic dopamine transmission dramatically increases sensitivity to the rewarding effects of nicotine in the ventral tegmental area. AB - Nicotine produces rewarding and aversive motivational effects in humans and other animal species. Here, we report that the mammalian ventral tegmental area (VTA) represents a critical neural substrate for the mediation of both the rewarding and aversive properties of nicotine. We demonstrate that direct infusions of nicotine into the VTA can produce both rewarding and aversive motivational effects. While the rewarding effects of higher doses of nicotine were not attenuated by dopamine (DA) receptor blockade, blockade of mesolimbic DA signalling with either systemic or intra-nucleus accumbens (NAc) neuroleptic pretreatment potentiated the sensitivity to nicotine's rewarding properties over a three-order-of-magnitude dose range. Furthermore, the behavioural effects of lower doses of intra-VTA nicotine were reversed, switching the motivational valence of nicotine from aversive to rewarding. Our results suggest that blockade of mesolimbic DA signalling induced by neuroleptic medications may block selectively the aversive properties of nicotine, thus increasing the vulnerability to nicotine's rewarding and addictive properties by inducing a unique, drug-vulnerable phenotype. PMID- 12556909 TI - The Multi-Source Interference Task: validation study with fMRI in individual subjects. AB - Dorsal anterior cingulate cortex (dACC) plays critical roles in cognitive processing, but group-averaging techniques have generally been required to obtain significant dACC activation in functional neuroimaging studies. Development of a task that reliably and robustly activates dACC within individuals is needed to improve imaging studies of neuropsychiatric disorders and localization of dACC in normal volunteers. By combining sources of cognitive interference (Stroop, Eriksen and Simon) with factors known to increase dACC activity, the Multi-Source Interference Task (MSIT) maximally taxes dACC, making it possible to reliably activate dACC within individuals using functional magnetic resonance imaging (fMRI). In this study, eight normal adult volunteers performed the MSIT during fMRI. We compared fMRI responses and performance data between interference and control trials. Significant dACC activation (P < 1.7 x 10(-4)) was observed in all eight individuals and in the group-averaged fMRI data. In addition to dACC activation, group data also showed activation of presumably networked regions including dorsolateral prefrontal, premotor, and parietal cortices. The MSIT's reaction time interference effect (overall mean 312 +/- 61 ms) was up to 10 times greater than that of its component predecessors and temporally stable over hundreds of trials. The robustness, reliability and stability of the neuroimaging and performance data should make the MSIT a useful task with which to study normal human cognition and psychiatric pathophysiology. PMID- 12556910 TI - A chromosome 14 risk locus for simple phobia: results from a genomewide linkage scan. AB - We conducted a 10 centimorgan (cM) linkage genome scan in a set of American extended pedigrees ascertained through probands with panic disorder. Several anxiety disorders segregate in these families. In this article, we describe results for simple phobia from 14 of these families (including 129 subjects of whom 57 are affected). A total of 422 markers were genotyped. Multipoint lod score analyses (fully parametric and simple parametric models) and nonparametric analyses were completed using ALLEGRO. We observed significant linkage of simple phobia to chromosome 14 markers. The highest lod score under a fully parametric model was 3.17, at marker D14S75, under a dominant model. Under a fully parametric recessive model, the maximum lod score, also at D14S75, was 2.86. Analysis under a simple parametric model resulted in lod scores of 3.70 (dominant model) or 3.30 (recessive model). The highest Zlr score observed was 3.93 (P = 4.1 x 10(-5)). The Zlr score was >1 for an extensive region, >77 cM. In all, 12 of the 14 families studied provided positive or zero lod scores at marker D14S75 (dominant model). The homologous genomic region has been implicated by studies mapping quantitative trait loci for a mouse model of fear. The linkage peak may be regarded as highly promising, owing to the breadth of the peak, the convergence of results under different models of inheritance and different analysis methods, and the support from an animal model. This is the first genome scan linkage study for simple phobia, a common disorder that causes high morbidity in the US population. PMID- 12556911 TI - A novel CpG-associated brain-expressed candidate gene for chromosome 18q-linked bipolar disorder. AB - We previously identified 18q21-q22 as a candidate region for bipolar (BP) disorder and constructed a yeast artificial chromosome (YAC) contig map. Here we identified three potential CpG islands using CCG/CGG YAC fragmentation. Analysis of available genomic sequences using bioinformatic tools identified an exon of 3639 bp downstream of a CpG island of 1.2 kb containing a putative transcription initiation site. The exon contained an open reading frame coding for 1212 amino acids with significant homology to the SART-2 protein; weaker homology was found with a series of sulphotransferases. Alignment of cDNA sequences of corresponding ESTs and RT-PCR sequencing predicted a transcript of 9.5 kb which was confirmed by Northern blot analysis. The transcript was expressed in different brain areas as well as in multiple other peripheral tissues. We performed an extensive mutation analysis in 113 BP patients. A total of nine single nucleotide polymorphisms (SNPs) were identified. Five SNPs predicted an amino acid change, of which two were present in BP patients but not in 163 control individuals. PMID- 12556912 TI - Association of D4 dopamine receptor gene and serotonin transporter promoter polymorphisms with infants' response to novelty. AB - Effects of DRD4 and 5-HTTLPR length polymorphisms have been reported on neonatal and infant temperament as well as adult personality traits. The 7-repeat form of the DRD4 III exon VNTR polymorphism has been associated with childhood ADHD, and recently we have reported its link with attachment disorganization in a nonclinical population of infants. Here, we report associations of these polymorphisms with infant temperament at 12 months of age. Maternal accounts of temperament and observed response to novelty were investigated for 90 infants, who were independently genotyped for the DRD4 III exon, and for 5-HTT-linked promoter region length polymorphisms. Maternal rating of temperament was not affected by these polymorphisms, but we found combined genotype effects for infants' observed responses to a novel, anxiety-provoking stimulus: the appearance of, and approach by, a stranger. Infants with at least one copy of both the 7-repeat DRD4 allele and the long variant of 5-HTTLPR (7(+), l/l&l/s) responded with significantly less anxiety than infants with other genotypes. However, infants with the 7-repeat DRD4 allele and homozygous for the short form of 5-HTTLPR (7(+), s/s) showed more anxiety and resistance to the stranger's initiation of interaction. These genotype effects were not redundant with the previously reported association between the 7-repeat DRD4 allele and disorganized attachment behavior. Although both temperament and attachment behavior were affected by the DRD4 repeat polymorphism, the effect on temperament measures was modified by the infants' 5-HTTLPR genotype. PMID- 12556913 TI - The serotonin 5-HT1B receptor gene and attention deficit hyperactivity disorder. AB - Recent research has suggested that serotonin, in addition to dopamine, may be involved in the development of attention deficit hyperactivity disorder (ADHD). Serotonin regulates dopaminergic neurotransmission in some areas of the brain via several 5-HT receptors including 5-HT1B. Animal studies have suggested the involvement of the 5-HT1B receptors in locomotor behaviour. For these reasons, we hypothesized that the 5-HT1B receptor gene may be a good candidate for genetic studies of ADHD. We tested for linkage disequilibrium between the 5-HT1B G861C polymorphism and ADHD in 115 families using the transmission disequilibrium test (TDT). We found evidence for a trend towards excess transmission of the 861G allele (chi(2) = 2.91, P = 0.09) that when further analysed for parental allele transmissions exhibited significantly greater paternal transmission of the G allele (chi(2) = 4.80, P = 0.03) to the affected child. Although preliminary, results from this study provide additional evidence that serotonin genes may be important risk factors for the development of ADHD. PMID- 12556914 TI - Mutational analysis of the nicotinic acetylcholine receptor alpha 4 subunit gene in attention deficit/hyperactivity disorder: evidence for association of an intronic polymorphism with attention problems. AB - Recent studies suggest the presence of genetically distinct subtypes of attention deficit/hyperactivity disorder (ADHD) and that attention problems can be treated with receptor subtype selective nicotine agonists. In this study, individuals with two independent familial subtypes of ADHD defined by latent class analysis were systematically screened for sequence variations in the coding regions and intron/exon junctions of the nicotinic acetylcholine receptor alpha 4 subunit gene (CHRNA4). Common polymorphisms were used for transmission disequilibrium test (TDT) analyses. A significant association was found for a 5' intron 2 single nucleotide polymorphism and severe inattention problems (P = 0.007, effect size = 4, 95% CI 1.3-14.1). The location of the polymorphism is compatible with it affecting pre-mRNA stability or splicing. PMID- 12556915 TI - D1 receptor alleles predict PET metabolic correlates of clinical response to clozapine. AB - A goal of pharmacogenetics is to clarify associations between allelic variation and risk factors in psychiatric illness. We report changes in regional brain metabolism based on dopamine alleles. Treatment-resistant schizophrenic subjects were positron emission tomography scanned with 18F-fluorodeoxyglucose after 5 weeks each of placebo and clozapine treatment. Significant regional brain metabolic effects were found for the D1 receptor genotypes (P < 0.05), adjusted for multiple comparisons. Metabolic decreases for the 2,2 genotype but not the 1,2 genotype were observed in all major sectors of the brain, with the exception of the ventral parts of the caudate and putamen. Frontal, temporal, parietal, and occipital neocortices showed decreased metabolism as did the cingulate juxta allocortex and the parahippocampal allocortex. Decreases were also observed in the thalamus, amygdala, and cerebellum bilaterally. No significant metabolic differences by genotype were observed for D3, 5HT(2A), and 5HT(2C) polymorphisms. In terms of clinical response, the DRD1 2,2 genotype significantly improved with clozapine treatment, demonstrating a 30% decrease in the Brief Psychiatric Rating Scale positive symptoms in contrast to a 7% worsening for the 1,2 genotype (P < 0.05). In this preliminary study, brain metabolic and clinical response to clozapine are related to the D1 receptor genotype. PMID- 12556916 TI - HLA and genetic susceptibility to sleepwalking. AB - HLA-DQB1 typing was performed in 60 Caucasian subjects with sleepwalking (SW) disorder and their families and 60 ethnically matched subjects without any diagnosed sleep disorder. A total of 21 sleepwalkers (35.0%) were DQB1*0501 positive vs eight (13.3%) controls (P = 0.0056; odds ratio = 3.5, 95% CI = 1.4 8.7). The family data for all HLA subtypes were further assessed for allelic association with SW using the transmission-disequilibrium test. A significant excess transmission was observed for DQB1*05 and *04 alleles in familial cases, strongly suggesting that a DQB1 polymorphic amino acid might be more tightly associated than any single allele. Sequence screening revealed that Ser74 in the second exon shared by all DQB1*05 and *04 was 20 times transmitted against 4 times non-transmitted (P = 0.001) in familial cases of SW. Thus, together with narcolepsy and REM sleep behavior disorder, these findings suggest that specific DQB1 genes are implicated in disorders of motor control during sleep. PMID- 12556917 TI - Association between a dinucleotide repeat polymorphism of the estrogen receptor alpha gene and personality traits in women. AB - Estrogens are known to play a key role in the regulation of various aspects of behavior. In order to study the potential contribution of genetic variation in the estrogen receptor (ER) alpha to specific personality traits, we investigated a repeat polymorphism in the ER alpha gene in 172 42-year-old women who had been assessed using the Karolinska Scales of Personality (KSP). Based on the hypothesis that there is a relationship between the length of a repeat polymorphism and gene function,(1) the alleles were divided into two groups: short and long. In order to elucidate the possible influence of the ER alpha gene on the different aspects of personality measured by means of the KSP, the possible association between this gene and four different factors ('neuroticism', 'psychoticism', 'non-conformity', and 'extraversion') was analysed. 'Neuroticism', 'psychoticism', and 'non-conformity' all appeared to be associated with the ER alpha gene. After correction for multiple comparisons by means of permutation analysis, the associations with the factor 'non-conformity'- including the subscales 'indirect aggression' and 'irritability'--and the factor 'psychoticism'--including the subscale 'suspicion'--remained significant. The results suggest that the studied dinucleotide repeat polymorphism of the ER alpha gene may contribute to specific components of personality. PMID- 12556918 TI - Truth is a corrected error. PMID- 12556919 TI - Perinatal substance abuse intervention in obstetric clinics decreases adverse neonatal outcomes. AB - OBJECTIVE: To evaluate the effect of Early Start, a managed care organization's obstetric clinic-based perinatal substance abuse treatment program, on neonatal outcomes. STUDY DESIGN: Study subjects were 6774 female Kaiser Permanente members who delivered babies between July 1, 1995 and June 30, 1998 and were screened by completing prenatal substance abuse screening questionnaires and urine toxicology screening tests. Four groups were compared: substance abusers screened, assessed, and treated by Early Start ("SAT," n=782); substance abusers screened and assessed by Early Start who had no follow-up treatment ("SA," n=348); substance abusers who were only screened ("S," n=262); and controls who screened negative ("C," n=5382). RESULTS: Infants of SAT women had assisted ventilation rates (1.5%) similar to control infants (1.4%), but lower than the SA (4.0%, p=0.01) and S groups (3.1%, p=0.12). Similar patterns were found for low birth weight and preterm delivery. CONCLUSION: Improved neonatal outcomes were found among babies whose mothers received substance abuse treatment integrated with prenatal care. The babies of SAT women did as well as control infants on rates of assisted ventilation, low birth weight, and preterm delivery. They had lower rates of these three neonatal outcomes than infants of either SA or S women. PMID- 12556920 TI - Site of care influences breastmilk feedings at NICU discharge. AB - OBJECTIVE: To determine if site of care influences rates of breastmilk use in neonatal intensive care units (NICUs). STUDY DESIGN: Subjects included all admissions to 124 NICUs between 1/1/1999 and 12/31/2000 who were subsequently discharged home. We reviewed discharge feeding data collected during the routine provision of care, and used stepwise logistic regression analysis to identify which factors were independently associated with a neonate being discharged on some breastmilk versus none. RESULTS: We studied 42,891 neonates; 21,327 (49.7%) were sent home receiving some breastmilk and 21,564 (50.3%) were not. The significant variables associated with being discharged on breastmilk were more mature gestational age, white race, maternal age, parents being married, and site of care. Site of care remained significant even when adjusted for other variables. CONCLUSION: Neonates admitted to NICUs are often discharged on feedings that do not include breastmilk. Beyond previously identified risk factors, site of care significantly influences this occurrence. PMID- 12556921 TI - The sy-fi study: a randomized prospective trial of synchronized intermittent mandatory ventilation versus a high-frequency flow interrupter in infants less than 1000 g. AB - OBJECTIVE: Mechanical ventilation has significantly decreased mortality for preterm infants with respiratory distress syndrome. However, the barotrauma associated with mechanical ventilation is responsible for the development of bronchopulmonary dysplasia (BPD) in 20% to 90% of these infants, depending on gestational age. Recent studies suggest that high-frequency ventilation is associated with better gas exchange and less barotrauma. The purpose of this study is to determine if high-frequency ventilation, using the flow interrupter, reduces the incidence of BPD in the ELBW infant. STUDY DESIGN: Parental consent was obtained prior to mechanical ventilation. Infants less than 1000 g without significant congenital anomalies were eligible for study entry. Infants were randomized, prior to the onset of mechanical ventilation, to receive either synchronized intermittent mandatory ventilation (SIMV) or high-frequency flow interruption. Both ventilator methods were obtained using the Infant Star Ventilator (Mallinckrodt, St. Louis, MO). Data were collected prospectively on all study entrants and analyzed using SigmaStat Software. RESULTS: Forty-six infants, from two centers, were enrolled in this study. There was no significant difference in the ventilator days or BPD, defined as oxygen requirement at 36 weeks postmenstrual age, between the high-frequency ventilation and SIMV groups. PMID- 12556922 TI - Early and late postnatal identification of isolated lenticulostriate vasculopathy in preterm infants: associated findings. AB - OBJECTIVES: To determine the incidence, possible etiologies, and neurodevelopmental outcome of premature infants (<35 weeks) with isolated lenticulostriate vasculopathy (LSV). STUDY DESIGN: In a retrospective case control design, we reviewed the medical records of all premature infants who were admitted to our neonatal intensive care unit between 1996 and 2000. RESULTS: The prevalence of LSV was 4.6% (21 of 453). Patients with late LSV (detected after 10 days of age) had less exposure than controls to prenatal steroids [42.8% (6 of 14) vs. 92.8% (13 of 14), respectively; p<0.01], and prenatal antibiotics [42.8% (6 of 14) vs. 85.7% (12 of 14), respectively; p=0.01]. Fifty-seven percent (8 of 14) of patients with late LSV had a low Apgar score vs. 14.2% (2 of 14) of the control group (p=0.01). Patients with LSV also had more muscle tone abnormalities than controls at 6 months of age [33.3% (5 of 15) vs. 5.2% (1 of 19), respectively; p=0.03]. CONCLUSION: Patients with late LSV have less exposure to antenatal steroids and antibiotics, lower Apgar scores, and abnormal muscle tone at 6 months of age. PMID- 12556923 TI - Echocardiographic evaluation of umbilical venous catheter placement. AB - OBJECTIVE: To compare techniques for guiding and confirming placement of umbilical venous catheters (UVCs) using two-dimensional echocardiography. STUDY DESIGN: Fifty-three newborns admitted to our neonatal intensive care unit who required an UVC or who were transferred within 24 hours of UVC placement at a referring hospital were studied. UVC position was assessed by antero-posterior (AP) chest radiography (CXR), lateral CXR, and oxygenation data. The accuracy of the above techniques was compared to echocardiography with saline contrast injection. RESULTS: Echocardiography revealed that UVCs were located ideally at the right atrial/inferior vena cava junction in only 12 (23%) of 53 patients. Twenty-four (45%) were incorrectly positioned in the left atrium. The sensitivity and specificity of AP CXR in evaluating inappropriate UVC position were 32% and 89%, respectively. Lateral CXR and thoracic level on AP CXR did not predict accurately catheter position. UVC pO(2) data were not useful in excluding left atrial placement. CONCLUSION: Current methods to determine insertion length and confirm location of UVCs are not adequate. Echocardiography should be considered to confirm correct placement of UVCs. PMID- 12556924 TI - Temporal patterns of caregiving for preterm infants indicate individualized developmental care. AB - OBJECTIVE: Individualized attention for premature infants was assessed from temporal patterns of caregiving at three very different regional NICUs. STUDY DESIGN: Continuous time-lapse video recording of 95 premature infants under 1500 g was made over three 24-hour periods when they were 33 weeks' and 35 weeks' conceptional age. STATISTICAL ANALYSIS: Analysis of variance was used to compare caregiving measures within and across hospitals. RESULTS: The tapes were scored in 5-minute epochs for periods when the infants were in and out of the crib, and when intervention/caregiving was occurring. Despite other differences, the three hospitals did not differ with respect to demographic and risk characteristics of the infant subjects. In all three hospitals, there was more out-of-crib and more caregiving during the daytime than at night; caregiving time showed individual differences over day and night periods; developmental changes in caregiving were observed over the 2 weeks; and more caregiving time was devoted to infants who were at greater risk. CONCLUSION: Temporal patterns of caregiving reliably indicate individual differences, with sensitive and responsive caregiving common to typical NICU nursing environments. PMID- 12556925 TI - Patient compliance with the first newborn visit appointment. AB - OBJECTIVE: To analyze patient compliance regarding the first newborn visit after hospital discharge. STUDY DESIGN: We selected at random 640 charts over a 1-year period from two community hospitals of healthy term newborns who were cared for by four practices in Kalamazoo, MI. We collected demographic data from the hospital chart and recorded the discharge order for time of posthospital follow up. We then accessed the individual clinics' appointment logs to determine when the patient came for the first visit. Lateness was defined as appearance for appointment >24 hours after the time stated on the hospital discharge order. We studied patient lateness with respect to the clinic, maternal age, insurance status, and distance from the patient's home to the medical facility, using both univariate and multivariate analyses. RESULTS: We demonstrated a significant difference in lateness of the first appointment between Medicaid and non-Medicaid patients in both the univariate (p<0.001) and multivariate (p=0.0003) analyses. We also demonstrated significant differences in the univariate analysis in patient lateness among the different practices (p<0.001) as well as lateness with regard to maternal age (p=0.0009). We did not attain significance in either analysis for patient lateness with regard to distance of home from the medical facility, or the hospital in which the baby was born. CONCLUSION: We demonstrated a need to improve upon newborn follow-up from the hospital and suggest strategies for other communities to analyze the process and improve upon patient and practice compliance with that first appointment. PMID- 12556926 TI - Functional outcome at adolescence for infants less than 801 g birth weight: perceptions of children and parents. AB - OBJECTIVE: To compare functional, emotional, and academic status of adolescents who had been extremely low birth weight (ELBW) with those who were full term. STUDY DESIGN: Twenty-six adolescents who were born in 1983 to 1984 at less than 801 g birth weight were compared with 26 adolescents born at term. Adolescent perceived status was assessed using the Behavior Assessment System for Children (BASC-SR) and the Self-Perception Profile. Parents' perceptions were assessed using BASC Parent Report, Family Impact Questionnaire, and Functional Status Measure. Health status was obtained through written requests to primary care providers and parent interviews. Academic and attendance information was obtained through school records. RESULTS: Compared to the term adolescents, the ELBW group had significantly lower function related to health status, increased need for special education, and tended to score lower on academic performance measures. However, ELBW adolescents' perceptions regarding their scholastic, athletic, or social competencies did not differ from the term group or national normative data. Parents of ELBWs reported lower adaptive skills, but no other behavioral differences from the term group. CONCLUSION: ELBW survivors and their parents perceived outcome at adolescence positively in spite of evident limitations. Mechanisms that ELBW families use to cope may be important determinants for successful, long-term outcome. PMID- 12556927 TI - Reflections on errors in neonatology: I. The "Hands-Off" years, 1920 to 1950. AB - This series discusses errors in neonatology since the 1920s. Three historical periods are defined: the "Hands-Off" years, 1920 to 1950; the "Heroic" years, 1950 to 1970; and the "Experienced" years, 1970 to 2000. In this article, the "Hands-Off" years, we discuss lowered thermal environment, supplemental oxygen, initial thirsting and starving, synthetic vitamin K, SMA formula, and diaper markings. PMID- 12556928 TI - A case of Wilson-Mikity syndrome with high serum KL-6 levels. AB - KL-6 and surfactant protein D (SP-D) serum concentrations are known to rise in adult patients with various types of interstitial pneumonia. We evaluated the time course of serum KL-6, SP-D, and lactate dehydrogenase (LDH) levels in an infant with Wilson-Mikity syndrome. In this case, serum KL-6 levels correlated with clinical symptoms better than serum SP-D and LDH levels. The findings of this case suggest that high serum levels of KL-6 show activity of interstitial lung disease of Wilson-Mikity syndrome. PMID- 12556929 TI - Urinary tract infection with Trichomonas vaginalis in a premature newborn infant and the development of chronic lung disease. AB - We report a case of a low-birth-weight infant with an infection of the urinary tract with Trichomonas vaginalis, who later developed cystic chronic lung disease suggestive of Wilson-Mikity syndrome. Although she had mild respiratory distress syndrome at birth, the extent of the chronic lung disease was out of proportion to the initial illness. We speculate that maternal infection with this organism may have resulted in an inflammatory response that led to its development. PMID- 12556930 TI - Familial glucocorticoid deficiency type 2 in two neonates. AB - Familial glucocorticoid deficiency (FGD) is a rare autosomal recessive disorder characterized by glucocorticoid deficiency, elevated plasma adrenocorticotropic hormone (ACTH) levels and preserved aldosterone/renin secretion. Adrenocorticotropic receptor mutations occur in about 40% of patients (FGD type 1), whereas the rest of the cases are associated with a normal receptor (FGD type 2). More than 50 cases have been reported in the literature so far. We report two cases of type 2 FGD from two related families who presented in the newborn period with varying clinical features. Direct sequencing of the genomic DNA of the patients failed to reveal mutations or other defects in the coding sequence of the ACTH receptor. Linkage analysis excluded mutations on the MC2-R gene outside the coding region. To our knowledge, these are the first two cases of FGD reported from the Middle East. PMID- 12556931 TI - Successful percutaneous retrieval of an intracardiac catheter fragment in an 800 g premature infant. AB - OBJECTIVE: To present the case report of a successful retrieval of an intracardiac catheter in an 800 - g premature infant. STUDY DESIGN: Case report. RESULTS: This report presents the technique for successful percutaneous removal of a retained intracardiac catheter fragment in an 800 - g infant. CONCLUSION: This study demonstrates that this procedure can be successfully performed in extremely small infants. PMID- 12556932 TI - Pancreatic pseudocysts in pregnancy: a case report and review of the literature. AB - BACKGROUND: Pancreatic pseudocyst in pregnancy is a rare condition whose management is not standardized. STUDY DESIGN: We combine one case report with nine others published in the literature since 1980. The cases are compiled to provide a descriptive review of this condition. RESULTS: The natural history of pancreatic pseudocysts in pregnancy appears similar to that in nongravid patients. Hyperlipidemia is overrepresented as a cause of pancreatic pseudocysts in pregnancy, causing more cases than alcoholic and biliary pancreatitis combined. Seventy-five percent of cases of known parity was primaparous. While in some cases percutaneous or endoscopic drainage was performed antepartum, most patients were conservatively managed until delivery. Despite two cases of successful vaginal delivery, cesarian section may be preferable for large pseudocysts to avert rupture. PMID- 12556933 TI - Mucopolysaccharidosis Type VII presenting with isolated neonatal ascites. AB - Mucopolysaccharidosis Type VII (MPS VII) is a lysosomal storage disease caused by a deficiency of the enzyme, beta-glucuronidase. MPS VII has a wide variation in phenotypic expression, including presentation in the neonatal period with nonimmune hydrops fetalis. We report a neonate with MPS VII who initially presented with marked isolated ascites not associated with hydrops fetalis. This appears to be a novel finding in patients with MPS VII. PMID- 12556934 TI - Fetal hydrocolpos leading to Pierre Robin sequence: an unreported effect of oligohydramnios sequence. AB - The presence of distal atretic vagina causing accumulation of fluid and mucus secretions in the proximal vaginal cavity resulted in fetal hydrocolpos. Obstructive uropathy developed gradually because of direct compression of hydrocolpos on bilateral lower ureters, resulting in oligohydramnios from decreased urine formation. Oligohydramnios inhibited normal mandibular development with resulting cleft palate and glossoptosis (Pierre Robin Sequence). The development of sequence of events in this case indicates Pierre Robin Sequence as another effect of Oligohydramnios Sequence arising out of deformational forces acting on cranio-facial structures. PMID- 12556935 TI - Neonatal tuberculosis associated with shock, disseminated intravascular coagulation, hemophagocytic syndrome, and hypercalcemia: a case report. AB - We report on a female infant with disseminated tuberculosis who presented with clinical sepsis and disseminated intravascular coagulation starting at 14 days of age. Parenteral ofloxacin combined with streptomycin were used because the enteral route was not possible and intravenous isoniazid and rifampicin were not available. Rare complications including infection-associated hemophagocytic syndrome, hypercalcemia, and adrenal insufficiency were detected and successfully managed. PMID- 12556936 TI - Fibrin glue for persistent pneumothorax in neonates. AB - Fibrin glue was used to treat significant pneumothoraces persisting for an average of 10 days in eight newborns. Six of the eight infants had reduction or resolution of persistent air leak within 24 hours of therapy. Two infants received a second course of therapy for recurrences. Complications encountered were bradycardia requiring manual ventilation (N=2), significant hypercalcemia (N=2), diaphragmatic paralysis (N=2), pneumothorax (PTX) on the contralateral side (N=1), and localized tissue necrosis (N=1). Fibrin glue is an effective treatment for intractable PTX but has significant risks. PMID- 12556937 TI - Thrombotic thrombocytopenic purpura in a newborn. AB - This report describes a newborn who presented with hyperbilirubinemia and thrombocytopenia. The patient recovered after treatment with antibiotics, phototherapy, and a platelet transfusion. Analysis of the plasma von Willebrand factor-cleaving metalloprotease, ADAMTS13, revealed low protease activity in the patient and her two siblings, and a mild deficiency in both parents. These results confirmed the clinical suspicion of hereditary thrombotic thrombocytopenic purpura (TTP). Although most cases of thrombocytopenia and hyperbilirubinemia in the newborn period are caused by other causes, genetic deficiency of ADAMTS13 may not be as uncommon as previously believed. Early diagnosis may have important implications for the patients. PMID- 12556941 TI - Fermented milks: a historical food with modern applications--a review. AB - BACKGROUND: : This paper was presented at the symposium which was organized by Instituto Danone Mexico in May 2001, and it provides an overview of the current scientific knowledge on fermented milks concerning the historical developments, manufacturing stages, classification of such products, and nutritional aspects. Particular attention has been paid to the human health benefits associated with the consumption of these products, the use of probiotic starter cultures and their industrial applications, and the significance of using a trained sensory panel for the evaluation of probiotic fermented milks made with different commercial blends of starter cultures. The paper also highlights the future research areas for the exploitation of starter microflora (Lactobacillus, Bifidobacterium and Enterococcus species) in fermented milk products. CONCLUSION: : This review indicates that the complex metabolism of the starter cultures is well established; however, more information is still needed on specific microbial metabolites such as polymerization of milk sugars for the production of exopolysaccharides and the modification of the milk peptides and secretion of bacteriocins. More clinical studies are still required to establish the 'functional' health benefits of probiotic fermented milks to humans. PMID- 12556942 TI - The role of fermented milk in complementary feeding of young children: lessons from transition countries. AB - Probiotic bacteria are used for production of fermented dairy products. The use of probiotic bacteria has the potential to replenish the natural intestinal flora of the body. These bacteria competitively inhibit the growth and colonization of pathogenic bacteria. Breastmilk is the best food for babies, also from a probiotic point of view. Human milk, in fact, contains many substances that stimulate the growth of bifidobacteria in vitro and in the small intestine of infants. Improvement of lactose digestion and avoidance of symptoms of intolerance in lactose malabsorbers are the most profoundly studied health relevant effects of fermented milk. In fact fermented milks are nutritionally similar to unfermented milk, except that some of lactose is broken down to glucose and galactose. The role of fermented milk in complementary feeding and in particular for the prevention of anaemia is an innovative theme, recently focused. Iron deficiency in infants and young children is widespread and has serious consequences for child health. Prevention of iron deficiency should therefore be given high priority. The too-early introduction of unmodified cow's milk and milk products is an important nutritional risk factors for the development of iron-deficiency anaemia. Fermented milks represent an excellent source of nutrients such as calcium, protein, phosphorus and riboflavin. During the fermentation of milk, lactic acid and other organic acids are produced and these increase the absorption of iron. If fermented milk is consumed at mealtimes, these acids are likely to have a positive effect on the absorption of iron from other foods. PMID- 12556943 TI - Interaction of lactic acid bacteria with the gut immune system. AB - Health claims of lactic acid bacteria (LAB) used in functional foods and pharmaceutical preparations are based on the capacity of these microorganisms to stimulate the host immune system. In this study, the antigenic effect of LAB (Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus) on the gut immune system of BALB/c mice was evaluated. A dose-dependent increase of the Bcl2 protein was observed with all LAB assayed. Furthermore, the analysis of cytokine-producing cells in the lamina propria of gut showed that TNFalpha and INFgamma values, determined in macrophages cultured from Peyer patches, were enhanced for all the LAB assayed. An important increase of interleukins IL-10 and IL-4 was observed mainly in mice fed with Lactobacillus delbrueckii ssp. bulgaricus or Lactobacillus casei, while a significant induction of IL-2 and IL-12 was only observed with L. acidophilus (P<0.01). These effects were dose dependent. The role of produced cytokines in the balance Th1/Th2 was determined by a systemic antibody response against parenterally injected ovoalbumin. L. casei, L. delbrueckii ssp. bulgaricus and L. acidophilus enhanced the IgG1 response favouring Th2 balance, while L. acidophilus also increased the IgG2a response inducing Th1 balance. S. thermophilus did not influence the balance Th1/Th2. Our studies showed that lactic acid bacteria induce distinct mucosal cytokine profiles showing different adjuvant capacity among them. Thus, selection of probiotic strain with immunological properties must be well defined to influence cytokine expression that favour the claimed immune response. PMID- 12556944 TI - The effect of fermented milk on interferon production in malnourished children and in anorexia nervosa patients undergoing nutritional care. AB - For several years cytokine production has been associated with infections but it was not suspected that some types of food could also induce cytokines, even in a state of non-infection. Lactic bacteria can induce interferon (IFN) production in human healthy subjects, thus, a better protection against infections would be expected. Therefore, we planned to evaluate the effect of two diets including yoghurt or milk on IFN-gamma production during nutritional recovery in two different situations of malnutrition: (1) children with diarrhoea; and (2) patients with anorexia nervosa (AN). Both the diet including yoghurt of that including milk seemed to increase IFN-gamma production at the end of nutritional recovery in the malnourished children with diarrhoea. The significance of interferon production and the lymphocyte subset increase should be explored to know if a better resistance against pathogens is related to them. Regulation of intestinal absorption and moderate stimulation of interferon production make the yoghurt-based diet a good choice in the nutritional care of children. In the same way, an increase in the IFN-gamma production was observed in AN patients consuming yoghurt. This increase of IFN-gamma production could be considered a biological marker to detect the effect of probiotics on the immune response, especially in the improvement of a deficient nutritional status. PMID- 12556945 TI - Role of microecology in chronic inflammatory bowel diseases. AB - Inflammatory bowel diseases (IBD) are chronic conditions of unknown etiology. Current therapy mitigates the severity of acute bouts of mucosal inflammation but an eradication therapy is lacking. Growing incidence of IBD is associated with social development. Epidemiology suggests a relationship between the establishment of the individual gut flora and the risk of developing IBD. Patients show an impaired tolerance towards commensal bacteria of the resident flora. Unrestrained activation of the intestinal immune system against some commensal bacteria appears to be responsible for the characteristic relapsing course of these diseases. Wide-spectrum antibiotic therapy reduces bacterial load and mitigates intestinal inflammation in human IBD and in animal models. Current research aims at the identification of probiotics for bacterial antagonism therapies. Probiotics are living microorganisms which upon ingestion in certain numbers exert health benefits beyond inherent basic nutrition. Colonization with a Lactobacillus reuteri strain can prevent the development of colitis in genetically susceptible mice. Other studies have used a bacterium genetically engineered to secrete the antiinflammatory cytokine IL-10 and demonstrated a therapeutic effect in animal models of colitis. Moreover, some probiotics may naturally exhibit antiinflammatory properties when interacting with the human gut mucosa. Prebiotics such as inulin have also been shown to prevent colonic inflammation in animal models. Preliminary clinical trials with probiotics in IBD are encouraging. Probiotics offer a valuable tool for the prevention and control of inflammatory bowel diseases. PMID- 12556946 TI - Asthma and allergic diseases: is there a downside to cleanliness and can we exploit it? AB - When hay fever was first described in the early 19th century it was an uncommon disorder. Since then asthma, allergic rhinitis and eczema, have become common conditions particularly in industrialised western economies. International prevalence studies reveal wide variation in the prevalence of asthma, and allergic disease, but confirm this view although the studies also show these diseases to be by no means rare in most countries. The reasons remain unclear but the "hygiene hypthesis" postulating an inverse relationship between hygiene, in its broadest sense, and allergic disease, fits some of the epidemiology of these diseases and has an associated immunological hypthesis to support it. Recent studies suggest that many hygiene related factors may influence immune development in favour of an allergic phenotype. Antibiotics in the first of life have been associated with increased risks of allergic disease in later childgood, and farming exposures to protection. Interest in hte role of bowel flora in modifying immune development has been suggested as an explanation for the risk associated with antibiotic exposure and has led to studies exploring the effects of modifying bowel flora with probiotics both to influence established allergic disease and to prevent it. The challenge is to continue to reap the enormous benefits that accrued from modifying infectious disease by both public and individual health strategies but at the same time convincing the immune system that it has been exposed to them. PMID- 12556947 TI - Analysis of the intestinal microflora using molecular methods. AB - A large and complex bacterial community inhabits the distal intestinal tract of humans. This collection, known as the intestinal microflora, is dominated numerically by obligately anaerobic bacterial species. Many of these species have never been cultivated under laboratory conditions. Nucleic acid-based techniques now permit, however, the analysis of even the non-cultivable members of the bacterial community. Polymerase chain reaction (PCR) coupled with denaturing gradient gel electrophoresis (DGGE) provides a useful technique for comparisons of the composition of faecal or intestinal microfloras. PCR/DGGE has been shown to be useful in demonstrating changes that occur in the composition of the faecal microflora of infants administered antibacterial drugs. This research is important because treatment with oral antibiotics during the first 2 y of life has been identified as a predictor of subsequent atopic disease. The treatment of young children with broad spectrum oral antibiotics might produce perturbations in the composition of the intestinal microflora such that bacteria important in promoting Th1 mechanisms are depleted at a crucial age. This could result in Th2 dominance over Th1 immune responses to environmental antigens and an increased incidence of atopic disorders. PCR/DGGE provides a useful screening method to determine the impact of antibiotic treatment on the composition of the intestinal microflora of children and to identify the bacterial groups that are most affected. PMID- 12556948 TI - Fermentation, fermented foods and lactose intolerance. AB - Lactose (milk sugar) is a fermentable substrate. It can be fermented outside of the body to produce cheeses, yoghurts and acidified milks. It can be fermented within the large intestine in those people who have insufficient expression of lactase enzyme on the intestinal mucosa to ferment this disaccharide to its absorbable, simple hexose sugars: glucose and galactose. In this way, the issues of lactose intolerance and of fermented foods are joined. It is only at the extremes of life, in infancy and old age, in which severe and life-threatening consequences from lactose maldigestion may occur. Fermentation as part of food processing can be used for preservation, for liberation of pre-digested nutrients, or to create ethanolic beverages. Almost all cultures and ethnic groups have developed some typical forms of fermented foods. Lessons from fermentation of non-dairy items may be applicable to fermentation of milk, and vice versa. PMID- 12556949 TI - Feasibility studies to control acute diarrhoea in children by feeding fermented milk preparations Actimel and Indian Dahi. AB - The aim of this work was to study feasibility of diarrhoea control in children (6 months to 5 y of age) by feeding fermented milk preparations. The design used was a randomized controlled clinical trial and the study was carried out at the Delhi University College Hospital providing tertiary care, and a nearby community centre Nand Nagri, a resettlement colony in East Delhi. Children suffering from acute diarrhoea (75 patients from the hospital and 75 from the community) were allocated to three groups by double-blind technique. Group 1 was given a fermented milk, Actimel, containing 10(8) of each Lactobacillus casei DN-114001, Lactobacillus bulgaricus and Streptococcus thermophilus per gram. Group 2 was given Indian Dahi (Lf 40) containing 10(8) of each Lactococcus lactis, Lactococcus lactis cremoris and Leuconostac mesenteroides cremoris per gram. Group 3 was given ultra-heat-treated yoghurt preparation (no live bacteria). Actimel was also used as a starter to prepare the curd in order to study the preventive effect of diarrhoea in children in a community. In the hospital study Indian Dahi and Actimel administration reduced mean duration of diarrhoea by 0.3 and 0.6 day (P<0.001), respectively. The corresponding figures in the community study were 0.2 and 0.5 day (P<0.05), respectively. The families using Actimel as a starter showed a reduction in diarrhoeal morbidity episodes by 40% of the children tested in a 3 month follow-up. In conclusion, Actimel, fermented milk containing Lactobacillus casei DN-114001, and Indian Dahi can significantly reduce the duration of diarrhoea in children; the former preparation being superior. PMID- 12556950 TI - Review of the distribution of Kaposi's sarcoma-associated herpesvirus (KSHV) in Africa in relation to the incidence of Kaposi's sarcoma. AB - In the years before human immunodeficiency virus (HIV) infection, the incidence of Kaposi's sarcoma varied markedly across the African continent, and it was a disease primarily affecting men. In contrast, the evidence reviewed here shows that the causal virus-Kaposi's sarcoma associated herpesvirus (KSHV)-is prevalent in many African countries, including places where Kaposi's sarcoma was almost unknown before HIV, and that it is as common in women as in men. Therefore, the geographical distribution of Kaposi's sarcoma in Africa before the spread of HIV and its predominance as a disease affecting men are not a simple reflection of the distribution of KSHV. Since the epidemic of HIV in Africa, Kaposi's sarcoma has become relatively more frequent in women, and the incidence has increased in countries where it was previously rare, but where KSHV is prevalent, as well as in countries where it was already common. These changes point to a role for other (as yet unknown) factors in the aetiology of Kaposi's sarcoma that may have the most effect in the absence of concurrent HIV infection. PMID- 12556951 TI - Current applications and future direction of MR mammography. AB - Compared with triple assessment for symptomatic and occult breast disease, magnetic resonance mammography (MRM) offers higher sensitivity for the detection of multifocal cancer, which is important in selecting patients appropriately for breast-conserving surgery. It is an ideal tool for the screening of patients with a high risk of breast cancer or where there is axillary disease or nipple discharge and conventional imaging has not revealed the primary focus. Techniques are now available to biopsy lesions only apparent on MRM. MRM can differentiate scar tissue from tumour; therefore, it is useful in patients in which there is possible recurrent disease. Clinical and X-ray mammographic assessment of response to neoadjuvant chemotherapy may be unreliable because of replacement of the tumour with scar tissue. MRM can identify responders and nonresponders with more accuracy. It is the modality of choice for the assessment of breast implants for rupture with accuracy higher than X-ray mammography and ultrasound. Advances in both spatial and temporal resolutions, the imaging sequences employed, pharmacokinetic modelling of contrast uptake, the use of dedicated and now phased array breast coils, and gadolinium-based contrast agents have all played their part in the advancement of this imaging technique. Despite the limitations of patient compliance, scan-time and cost, this review describes how MRM has become a valuable tool in breast disease, especially in cases of diagnostic uncertainty. However, MRM must make the transition from research institutions into routine clinical practice. PMID- 12556952 TI - Docetaxel induction therapy in locally advanced squamous cell carcinoma of the head and neck. AB - Patients with locally advanced squamous cell carcinoma of the head and neck (SCCHN) are often treated with induction chemotherapy or chemoradiotherapy, but to date without major impact on survival. The combination of cisplatin-5 fluorouracil (5-FU) (PF) has been used as standard induction therapy; however, poor patient survival has stimulated investigation into new agents with potential activity in SCCHN. Docetaxel has significant single-agent activity in SCCHN and has been investigated in combination with PF regimens as induction therapy. The results of six phase II studies of docetaxel-PF regimens (TPF) as induction in locally advanced SCCHN patients are reviewed and reported. Consistently, high 2 year survival rates and overall response rates were demonstrated across the phase II trials in the range 42-82 and 71-100%, respectively. The toxicity profile seen with TPF-based regimens was acceptable. The primary toxicity was neutropenia, which together with gastrointestinal complaints accounted for the majority of adverse events. Given the encouraging phase II experience with TPF-based regimens, two large-scale phase III studies comparing TPF-based regimens with standard PF regimens are underway. The results have significant potential for validating the findings of the phase II studies, demonstrating improved survival and overall response of patients treated with docetaxel-based induction chemotherapy. PMID- 12556953 TI - Definitive chemoradiotherapy for patients with malignant stricture due to T3 or T4 squamous cell carcinoma of the oesophagus. AB - We retrospectively investigated the efficacy and feasibility of concurrent chemoradiotherapy for patients with severe dysphagia caused by oesophageal squamous cell carcinoma. Concurrent chemoradiotherapy was performed in 57 patients with T3 or T4 disease containing M1 lymph node (LYM) disease. Chemotherapy consisted of protracted infusion of 5-fluorouracil (5-FU) 400 mg m( 2) 24 h(-1) on days 1-5 and 8-12, combined with 2-h infusion of cisplatin (CDDP) 40 mg m(-2) on days 1 and 8. Radiation treatment at a dose of 30 Gy in 15 fractions of the mediastinum was administered concomitantly with chemotherapy. A course schedule with 3-week treatment and a 1 to 2-week break was applied twice, with a total radiation dose of 60 Gy, followed by two or more courses of 5-FU and CDDP. In all, 24 patients (42%) achieved a complete response, and the 3-year survival rate was 19%. Major toxicities were leukocytopenia and oesophagitis, and there were two (4%) treatment-related deaths. In contrast, 22 patients with T3 disease survived longer than 35 patients with T4 disease (P=0.001); however, the survival rate in 15 patients with M1 LYM disease did not differ significantly from that in 42 patients without M1 LYM disease (P=0.3545). Our results indicate that definitive chemoradiotherapy is potentially curative for locally advanced oesophageal carcinoma with malignant stricture. The efficacy and survival of patients treated with this regimen are related to the T factor. PMID- 12556954 TI - Feasibility of combination chemotherapy with cisplatin and etoposide for haemodialysis patients with lung cancer. AB - Cancer chemotherapy for haemodialysis patients has never been established. To elucidate the feasibility of cisplatin-based combination chemotherapy for haemodialysis patients with lung cancer, a dose escalation study was conducted. Five haemodialysis patients with lung cancer were treated with cisplatin and etoposide. A starting dose of 40 mg m(-2) of cisplatin on day 1 and 50 mg m(-2) of etoposide on days 1, 3 and 5 were administered as the first course for the first patient. Membrane haemodialysis was regularly performed three times a week and soon after the completion of therapy. By monitoring toxicity and pharmacokinetics data, the dose was escalated course by course and patient by patient. Dose escalation was completed for the first two patients resulting in full-dose chemotherapy consisting of 80 mg m(-2) of cisplatin on day 1 and 100 mg m(-2) of etoposide on days 1, 3 and 5. Multiple courses of the full-dose chemotherapy were administered to the other three patients. Toxicity was manageable and tolerable for all. Pharmacokinetics data were comparable to those from patients with normal renal function, except for potential long-lasting higher levels of free platinum in the renal insufficiency group. In conclusion, this standard-dose combination chemotherapy was feasible even for haemodialysis patients. PMID- 12556955 TI - Effectiveness and cost-effectiveness of prognostic markers in prostate cancer. AB - This paper demonstrates how economic modelling can be used to derive estimates of the cost-effectiveness of prognostic markers in the management of clinically localised and moderately graded prostate cancer. The model uses a Markov process and is populated using published evidence and local data. The robustness of the results has been tested using sensitivity analysis. Three treatment policies of 'monitoring' (observation), radical prostatectomy, or a selection-based management policy using DNA-ploidy as an experimental marker, have been evaluated. Modelling indicates that a policy of managing these tumours utilising experimental markers has an estimated cost per quality-adjusted life year (QALY) of pound 12 068. Sensitivity analysis shows the results to be relatively sensitive to quality-of-life variables. If novel and experimental markers can achieve specificity in excess of 80%, then a policy of radical surgery for those identified as being at high risk and conservative treatment for the remainder would be both better for patients and cost-effective. The analysis suggests that a radical prostatectomy treatment policy for the moderately graded tumours (Gleason grades -7) modelled in this paper may be inferior to a conservative approach in the absence of reliable prognostic markers, being both more costly and yielding fewer QALYs. PMID- 12556956 TI - Excessive annual BMI increase after chemotherapy among young survivors of testicular cancer. AB - Increased body mass index (BMI) is claimed to be a complication among survivors of testicular cancer (TCSs), especially after receiving cisplatin-based chemotherapy. This study compares changes in BMI (kg m(-2)) in TCSs with those observed in age-matched men from the population (controls). Associations between treatment, age and potential BMI changes were sought. In 1999, a survey was performed at the NRH of 444 unilaterally orchiectomised TCSs treated from 1980 to 1990. BMI at survey was recorded in each TCS. Information on principal treatment (surgery only: SURG; radiotherapy only: RAD; chemotherapy +/- surgery or radiotherapy: CHEM+/-) and pretreatment BMI was retrieved from the medical records. The age-matched controls had BMI measurements from population surveys from 1985 and 1996. The annual BMI increase was calculated based on the difference in the two BMI measurements divided by observation time. TCSs displayed a lower pretreatment mean BMI than the controls, whereas no difference was found post-treatment. However, the annual BMI increase in TCSs exceeded that of the controls (0.19 vs 0.15, P=1.4 x 10(-7)). The SURG and CHEM+/- groups showed the greatest annual BMI increase. The multiple regression analysis showed that young TC patients who received chemotherapy displayed an excessive annual BMI increase. Oncologists and young TCSs should be aware of the risk of excessive BMI increase, in particular, after the use of chemotherapy. PMID- 12556957 TI - Attitudes towards HPV testing: a qualitative study of beliefs among Indian, Pakistani, African-Caribbean and white British women in the UK. AB - This study examined attitudes to human papillomavirus (HPV) testing among a purposively selected sample of women from four ethnic groups: white British, African Caribbean, Pakistani and Indian. The design was qualitative, using focus group discussion to elicit women's attitudes towards HPV testing in the context of cervical cancer prevention. The findings indicate that although some women welcomed the possible introduction of HPV testing, they were not fully aware of the sexually transmitted nature of cervical cancer and expressed anxiety, confusion and stigma about HPV as a sexually transmitted infection. The term 'wart virus', often used by medical professionals to describe high-risk HPV to women, appeared to exacerbate stigma and confusion. Testing positive for HPV raised concerns about women's sexual relationships in terms of trust, fidelity, blame and protection, particularly for women in long-term monogamous relationships. Participation in HPV testing also had the potential to communicate messages of distrust, infidelity and promiscuity to women's partners, family and community. Concern about the current lack of available information about HPV was clearly expressed and public education about HPV was seen as necessary for the whole community, not only women. The management of HPV within cervical screening raises important questions about informed participation. Our findings suggest that HPV testing has the potential to cause psychosocial harm to women and their partners and families. PMID- 12556958 TI - Delayed infection, late tonsillectomy or adenoidectomy and adult leukaemia: a case-control study. AB - In a population-based case-control study among adults in Italy, of 261 lymphoid and 313 myeloid leukaemias and 1718 controls, a later age at adenoidectomy and tonsillectomy (after age 10 years) increased considerably the risk of lymphocytic (but not myeloid) leukaemia (odds ratio 4.2, 95% confidence interval 1.1-16.2). We propose that late infection is a proliferative stimulus for B-cells. PMID- 12556959 TI - Hormonal content and potency of oral contraceptives and breast cancer risk among young women. AB - Recent use of oral contraceptive pills is associated with a modest risk of breast cancer among very young women. In this US population-based case-control study, we evaluated whether the excess risk associated with recent oral contraceptive use is ubiquitous for all pill types or attributable to specific oral contraceptive preparations. Hormonal content and potency of combination oral contraceptives used for the longest duration within 5 years of interview for breast cancer cases aged 20-44 years (N=1640) were compared with age-matched community controls (N=1492). Women who recently used oral contraceptives containing more than 35 microg of ethinyl oestradiol per pill were at higher risk of breast cancer than users of lower dose preparations when compared to never users (respective relative risks of 1.99 and 1.27, P(trend)<0.01). This relationship was more marked among women <35 years of age, where risks associated with high- and low dose ethinyl oestradiol use were 3.62 and 1.91 (P(trend)<0.01), respectively. We also found significant trends of increasing breast cancer risk for pills with higher progestin and oestrogen potencies (P(trend)<0.05), which were most pronounced among women aged <35 years of age (P(trend)<0.01). Risk was similar across recently used progestin types. Our findings suggest that newer low potency/low oestrogen dose oral contraceptives may impart a lower risk of breast cancer than that associated with earlier high-potency/high-dose preparations. PMID- 12556960 TI - Glutathione S-transferase M1 and T1 genetic polymorphisms, alcohol consumption and breast cancer risk. AB - Alcohol consumption has been inconsistently associated with breast cancer risk. Recent studies suggest that genetic polymorphisms of glutathione S-transferases (GSTs) may modify this relation. To determine if breast cancer risk is associated with GSTM1 and GSTT1 genetic polymorphisms, and to evaluate the effect modification between GST genotypes and alcohol consumption in the risk of breast cancer, we conducted a case-control study in the state of Connecticut in the period 1998 and 2001. Cases were histologically confirmed, incident breast cancer patients in New Haven County, CT. Controls were randomly selected from women histologically confirmed to be without breast cancer. The study results show that, while GSTM1 genotypes were not associated with breast cancer risk, GSTT1 null genotype was associated with a significant 90% increased risk for postmenopausal women (OR=1.9, 95% CI 1.2-3.0). Analysis by GST genotypes and alcohol consumption shows that GSTM1A ever-drinking women had a 2.5-fold (OR=2.5, 95% CI 1.1-5.5) increased risk of breast cancer compared to the GSTM1A never drinkers, and the risk increases with duration and daily amount of alcohol consumption. Postmenopausal women with GSTT1-null genotype, who consumed a lifetime of >250 kg of spirit-equivalents, had an almost seven-fold increased risk (OR=6.8, 95% CI 1.4-33.9), and drinking commencing at younger ages appears to carry a higher risk. An OR of 8.2 (95% CI 1.2-57.4) was observed for those with GSTM1A, and GSTT1-null genotypes who had consumed a lifetime of >250 kg of spirit-equivalents. In conclusion, alcohol consumption may increase breast cancer risk among those who carry susceptible GST genotypes. PMID- 12556961 TI - Human papillomavirus types in invasive cervical cancer worldwide: a meta analysis. AB - This study investigated regional variations in the contribution made by different human papilloma (HPV) types to invasive cervical cancer (ICC). A total of 85 studies using polymerase chain reaction to estimate HPV prevalence in ICC were identified. Data on HPV prevalence were extracted separately for squamous cell carcinoma (SCC) and for adeno- and adenosquamous-carcinoma (ADC). A total of 10 058 cases (8550 SCC, 1508 ADC) were included in pooled analyses. The most common HPV types in ICC were, in order of decreasing prevalence, HPV16, 18, 45, 31, 33, 58, 52, 35, 59, 56, 6, 51, 68, 39, 82, 73, 66 and 70. In SCC, HPV16 was the predominant type (46-63%) followed by HPV18 (10-14%), 45 (2-8%), 31 (2-7%) and 33 (3-5%) in all regions except Asia, where HPV types 58 (6%) and 52 (4%) were more frequently identified. In ADC, HPV prevalence was significantly lower (76.4%) than in SCC (87.3%), and HPV18 was the predominant type in every region (37-41%), followed by 16 (26-36%) and 45 (5-7%). The overall detection of HPV DNA was similar in different regions (83-89%). A majority of ICC was associated with HPV16 or 18 in all regions, but approximately a quarter of all ICC cases were associated with one of 16 other HPV types, their distribution varying by region. PMID- 12556962 TI - Evidence for an association between cutaneous malignant melanoma and lymphoid malignancy: a population-based retrospective cohort study in Scotland. AB - We analysed the risk of cutaneous malignant melanoma (CM) occurring in patients following a diagnosis of non-Hodgkin's lymphoma (NHL) or chronic lymphatic leukaemia (CLL), and of NHL or CLL subsequently developing in CM survivors. Cohorts of patients with CM, NHL or CLL (index cancer) diagnosed between 1975 and 1997 were identified from the Scottish national cancer registry and followed through the registry for subsequent CM, NHL or CLL. The standardised incidence ratio (SIR) for each cancer was calculated and overall risk, risk in relation to gender and age at diagnosis of the index cancers and time from diagnosis of the index cancer to the diagnosis of the second malignancy were measured. There were 9385 CM patients, 4016 CLL patients and 13 857 NHL patients identified with an index cancer with 56 195, 14 450 and 44 999 person-years of follow-up, respectively. There was an increased risk of both CLL and NHL following a diagnosis of CM (SIR 2.3 and 1.5, respectively) and of CM following a diagnosis of CLL and NHL (SIR 2.3 and 2.1, respectively). The risk was statistically significantly increased for CLL developing in CM patients and for CM occurring in NHL survivors (P<0.05). This study supports an association between CM, CLL and NHL developing in the same patient. Immunosuppression, exposure to ultraviolet radiation and genetic factors may lead to a host environment that is conducive to the development of these malignancies. PMID- 12556963 TI - Cholecystectomy and the risk for developing colorectal cancer and distal colorectal adenomas. AB - Earlier work describes a modest association between cholecystectomy and the risk of colorectal cancer. We conducted a prospective study of 85 184 women, 36-61 years old, who had no history of cancer to evaluate whether known risk factors for colorectal cancer, including dietary history, that have not been controlled for in previous analyses can help explain the observed association. During 16 years of follow-up, 877 cases of colorectal cancer were documented and 1452 women who underwent endoscopy during the follow-up time were diagnosed with distal adenomas. After adjustment for age and other known or suspected risk factors, we found a significant, positive association between cholecystectomy and the risk of colorectal cancer (multivariate relative risk RR 1.21, 95% CI 1.01-1.46). The risk was highest for cancers of the proximal colon (RR 1.34, 95% CI 0.97-1.88) and the rectum (RR 1.58, 95% CI 1.05-2.36). However, we did not observe a significant association between cholecystectomy and distal colorectal adenomas. In this large prospective cohort study, a history of cholecystectomy appears to increase modestly the risk of colorectal cancer, even after adjustment for other colorectal cancer risk factors. PMID- 12556964 TI - Dietary acrylamide and cancer of the large bowel, kidney, and bladder: absence of an association in a population-based study in Sweden. AB - Recently, disturbingly high levels of acrylamide were unexpectedly detected in widely consumed food items, notably French fries, potato crisps, and bread. Much international public concern arose since acrylamide has been classified as a probable carcinogen, although based chiefly on laboratory evidence; informative human data are largely lacking. We reanalysed a population-based Swedish case control study encompassing cases with cancer of the large bowel (N=591), bladder (N=263) and kidney (N=133), and 538 healthy controls, assessing dietary acrylamide by linking extensive food frequency data with acrylamide levels in certain food items recorded by the Swedish National Food Administration. Unconditional logistic regression was used to estimate odds ratios, adjusting for potential confounders. We found consistently a lack of an excess risk, or any convincing trend, of cancer of the bowel, bladder, or kidney in high consumers of 14 different food items with a high (range 300-1200 microg kg(-1)) or moderate (range 30-299 microg kg(-1)) acrylamide content. Likewise, when we analysed quartiles of known dietary acrylamide intake, no association was found with cancer of the bladder or kidney. Unexpectedly, an inverse trend was found for large bowel cancer (P for trend 0.01) with a 40% reduced risk in the highest compared to lowest quartile. We found reassuring evidence that dietary exposure to acrylamide in amounts typically ingested by Swedish adults in certain foods has no measurable impact on risk of three major types of cancer. It should be noted, however, that relation of risk to the acrylamide content of all foods could not be studied. PMID- 12556965 TI - Mitochondrial DNA damage in non-melanoma skin cancer. AB - Mitochondrial DNA (mtDNA) damage, predominantly encompassing point mutations, has been reported in a variety of cancers. Here we present in human skin, the first detailed study of the distribution of multiple forms of mtDNA damage in nonmelanoma skin cancer (NMSC) compared to histologically normal perilesional dermis and epidermis. We present the first entire spectrum of deletions found between different types of skin tumours and perilesional skin. In addition, we provide the first quantitative data for the incidence of the common deletion as well as the first report of specific tandem duplications in tumours from any tissue. Importantly, this work shows that there are clear differences in the distribution of deletions between the tumour and the histologically normal perilesional skin. Furthermore, DNA sequencing of four mutation 'hotspot' regions of the mitochondrial genome identified a previously unreported somatic heteroplasmic mutation in an SCC patient. In addition, 81 unreported and reported homoplasmic single base changes were identified in the other NMSC patients. Unlike the distribution of deletions and the heteroplasmic mutation, these homoplasmic mutations were present in both tumour and perilesional skin, which suggests that for some genetic studies the traditional use of histologically normal perilesional skin from NMSC patients may be limited. Currently, it is unclear whether mtDNA damage has a direct link to skin cancer or it may simply reflect an underlying nuclear DNA instability. PMID- 12556966 TI - Accumulation of uPA-PAI-1 complexes inside the tumour cells is associated with axillary nodal invasion in progesterone-receptor-positive early breast cancer. AB - Both urokinase-like plasminogen activator (uPA) and its inhibitor plasminogen activator inhibitor (PAI-1), as well as uPA-PAI-1 complexes, have been identified as important prognostic factors in breast cancer. We have recently reported that the latter are identifiable inside breast cancer cells by means of immunohistochemistry. Using this technique, we have studied a series of 212 early (pT1) unifocal breast cancers and have correlated the expression of uPA-PAI-1 complexes, together with other clinical and biological features (histologic variety, histologic and nuclear grade, hormone receptors, Ki67 labelling index, c erb-B2-, p53- and CD44std-expression) with or without the occurrence of axillary node invasion. In a logistic regression model, looking for associations with axillary metastasis, we found a statistically significant interaction between the presence of uPA-PAI-1 complexes and progesterone receptor positivity (P=0.04). A final model showed that the presence of uPA-PAI-1 complexes was a determinant factor for axillary metastasis among women carrying tumours expressing progesterone receptors. In these cases, the presence of uPA-PAI-1 complexes carried with it a nearly 14-fold risk of axillary node invasion (P=0.009). These results may indicate that small, hormone-receptor-positive breast cancers (with a theoretical good prognosis) may carry an elevated risk of nodal involvement if accumulation of uPA-PAI-1 complexes is shown inside their tumour cells by means of immunohistochemistry. PMID- 12556967 TI - Independent prognostic value of angiogenesis and the level of plasminogen activator inhibitor type 1 in breast cancer patients. AB - Tumour angiogenesis and the levels of plasminogen activator inhibitor type 1 (PAI 1) are both informative prognostic markers in breast cancer. In cell cultures and in animal model systems, PAI-1 has a proangiogenic effect. To evaluate the interrelationship of angiogenesis and the PAI-1 level in breast cancer, we have evaluated the prognostic value of those factors in a total of 228 patients with primary, unilateral, invasive breast cancer, evaluated at a median follow-up time of 12 years. Microvessels were immunohistochemically stained by antibodies against CD34 and quantitated by the Chalkley counting technique. The levels of PAI-1 and its target proteinase uPA in tumour extracts were analysed by ELISA. The Chalkley count was not correlated with the levels of uPA or PAI-1. High values of uPA, PAI-1, and Chalkley count were all significantly correlated with a shorter recurrence-free survival and overall survival. In the multivariate analysis, the uPA level did not show independent prognostic impact for any of the analysed end points. In contrast, the risk of recurrence was independently and significantly predicted by both the PAI-1 level and the Chalkley count, with a hazard ratio (95% CI) of 1.6 (1.01-2.69) and 1.4 (1.02-1.81), respectively. For overall survival, the Chalkley count, but not PAI-1, was of significant independent prognostic value. The risk of death was 1.7 (1.30-2.15) for Chalkley counts in the upper tertile compared to the lower one. We conclude that the PAI-1 level and the Chalkley count are independent prognostic markers for recurrence free survival in patients with primary breast cancer, suggesting that the prognostic impact of PAI-1 is not only based on its involvement in angiogenesis. PMID- 12556968 TI - CpG island methylation status and mutation analysis of the RB1 gene essential promoter region and protein-binding pocket domain in nervous system tumours. AB - A series of 136 nervous system tumours were studied to determine the methylation status of the CpG island contained within the promoter region of the RB1 gene, as well as mutation analysis of the essential promoter region and exons 20-24 (and surrounding intronic regions) coding for the protein-binding pocket domain. Methylation of the RB1 CpG island was detected in 26 samples corresponding to nine glioblastomas, three anaplastic astrocytomas, one mixed oligo-astrocytoma, one ependymoma, two medulloblastomas, two primary central nervous system lymphomas, two neurofibrosarcomas, and six brain metastasis from solid tumours. No inactivating mutations were found within the RB1 promoter region, whereas one glioblastoma and one oligodendroglioma displayed similar sequence variations consisting of 12 and 8 base pair deletions at intron 21. These results suggest that RB1 CpG island hypermethylation is a common epigenetic event that is associated with the development of malignant nervous system tumours. PMID- 12556969 TI - Prognostic value of nuclear survivin expression in oesophageal squamous cell carcinoma. AB - Survivin, a new member of the family of apoptosis inhibitors, is expressed almost exclusively in proliferating cells, above all in cancers. Subcellular localisation and prognostic implications of the survivin protein have not yet been determined in oesophageal squamous cell carcinoma. The survival of 84 patients with oesophageal squamous cell carcinomas was correlated with the extent of immunohistochemical survivin expression in tumour cell nuclei. Tumours were scored positive when >5% cells stained positive. Patients were followed up for at least 5 years or until death. In normal oesophageal squamous cell epithelium, some cytoplasmic survivin expression was detected in the basal cells, whereas proliferating cells showed nuclear staining of survivin. Nuclear expression of survivin was also detected in 67 cancers (80%). The mean survival for patients of this group (28 months, range 20-36) was significantly less than that for patients without survivin expression in the tumour cell nuclei (108 months, range 62-154, P=0.003). Using univariate analysis, nuclear survivin expression (P=0.003), tumour depth (P=0.001), lymph node metastasis (P=0.003) and stage (P<0.001) were the best predictors of survival. In contrast, cytoplasmic survivin staining was noted in 53 (63%) tumours and had no prognostic relevance. In conclusion, the analysis of nuclear survivin expression identifies subgroups in oesophageal squamous cell cancer with favourable (survivin(-)) or with poor prognosis (survivin(+)). We suggest that the determination of nuclear survivin expression could be used to individualise therapeutic strategies in oesophageal squamous cell cancer in the future. PMID- 12556970 TI - NF-kappaB inhibition impairs the radioresponse of hypoxic EMT-6 tumour cells through downregulation of inducible nitric oxide synthase. AB - Hypoxic EMT-6 tumour cells displayed a high level of inducible nitric oxide synthase (iNOS) and an increased radiosensitivity after a 16 h exposure to lipopolysaccharide, a known activator of nuclear factor-kappaB (NF-kappaB). Both iNOS activation and radioresponse were impaired by the NF-kappaB inhibitors phenylarsine oxide and lactacystin. Contrasting to other studies, our data show that inhibition of NF-kappaB may impair the radioresponse of tumour cells through downregulation of iNOS. PMID- 12556971 TI - Staurosporine-induced apoptosis and hydrogen peroxide-induced necrosis in two human breast cell lines. AB - The use of apoptosis-inducing agents in the treatment of malignant cancer is increasingly being considered as a therapeutic approach. In this study, the induction of apoptosis and necrosis was examined in terms of temporal dose responses, comparing a malignant and nonmalignant breast cell line. Staurosporine (SSP)-induced apoptosis and H(2)O(2)-induced necrosis were evaluated by two cytotoxicity assays, neutral red (NR) and methyl-thiazolyl tertrazolium (MTT), in comparison with a differential dye uptake assay, using Hoechst33342/propidium iodide (Hoechst/PI). Confirmatory morphological assessment was also performed by routine resin histology and transmission electron microscopy. Cell viability was assessed over a 0.5-48 h time course. In nonmalignant HBL-100 cells, 50 nM SSP induced 100% apoptosis after a 48 h exposure, while the same exposure to SSP caused only 4% apoptosis in metastatic T47D cells. Although complete apoptosis of both cell lines was induced by 50 microM SSP, this effect was delayed in T47D (24 h) compared with HBL-100 (4 h). Results also showed that neither MTT or NR can distinguish between the modes of cell death, nor detect the early onset of apoptosis revealed by Hoechst/PI. PMID- 12556972 TI - Effect of a novel somatostatin analogue combined with cytotoxic drugs on human tumour xenografts and metastasis of B16 melanoma. AB - A novel somatostatin analogue, TT-232 (which inhibits the proliferation of various cell cultures and transplantable mouse tumours), was examined regarding its effect on human melanoma and lymphoma xenografts as a single treatment or in combination with DTIC (dacarbazine) and etoposide. TT-232 inhibited the growth of HT-18 melanoma xenografts, a dose of 5 mg kg(-1) being the most effective. Combination of 1 mg kg(-1) TT-232 with 30 or 60 mg kg(-1) DTIC (administered daily) resulted in a stronger inhibitory effect compared to TT-232 or DTIC as a single modality. Antimetastatic effect of TT-232 treatment combined with DTIC was studied using the B16 mouse melanoma muscle - lung metastasis model. The number of lung metastases of B16 melanoma could be decreased by the daily administration of 1 mg kg(-1) TT-232 or 60 mg kg(-1), but not of 30 mg kg(-1) DTIC. TT-232, combined with 30 or 60 mg kg(-1) DTIC decreased the lung metastasis number significantly lower than the control. Nearly 50% growth inhibition of HT-58 lymphoma was achieved by daily treatment with 1 mg kg(-1) TT-232. 5 mg kg(-1) etoposide, administered daily, resulted in a similar effect. The combination of 1 mg kg(-1) TT-232 and 5 mg kg(-1) etoposide was significantly more effective than TT-232 or etoposide as a single treatment. The very strong tumour growth inhibitory effect of 10 mg kg(-1) etoposide could even be increased by combination with TT-232. These experimental data suggest that TT-232 may be an effective new tool in the combination chemotherapy of malignant tumours like melanoma and lymphoma. PMID- 12556973 TI - Transformation induced by Ewing's sarcoma associated EWS/FLI-1 is suppressed by KRAB/FLI-1. AB - Ewing's sarcoma is a childhood bone tumour with poor prognosis, most commonly associated with a t(11;22)(q24;q12) reciprocal translocation that fuses the EWS and FLI-1 genes, resulting in the production of an aberrant chimeric transcription factor EWS/FLI-1. To elucidate the mechanisms by which EWS/FLI-1 mediates transformation in mouse models, we have generated a murine Ews/Fli-1 fusion protein. We demonstrate that this protein transforms fibroblast cells in vitro similar to human EWS/FLI-1 as demonstrated by serum and anchorage independent growth, the formation of tumours in nude mice and elevation of the oncogenic marker c-myc. Furthermore, transformation of these cells was inhibited by a specific repressor, KRAB/FLI-1. The KRAB/FLI-1 repressor also suppressed the tumorigenic phenotype of a human Ewing's sarcoma cell line. These findings suggest that the transformed phenotype of Ewing's sarcoma cells can be reversed by using the sequence-specific FLI-1-DNA-binding domain to target a gene repressor domain. The inhibition of EWS/FLI-1 is the first demonstration of the KRAB domain suppressing the action of an ETS factor. This approach provides potential avenues for the elucidation of the biological mechanisms of EWS/FLI-1 oncogenesis and the development of novel therapeutic strategies. PMID- 12556974 TI - Endoplasmic reticulum and Golgi apparatus are the preferential sites of Foscan localisation in cultured tumour cells. AB - Intracellular photosensitiser localisation significantly influences the mechanism of response to photodynamic therapy (PDT), since the primary sites of damage are closely related to the specific sensitiser distribution. Foscan subcellular localisation in the MCF-7 human adenocarcinoma cell line has been studied by means of confocal microscopy and microspectrofluorometry. The fluorescence topographic profiles recorded after cells costained with Foscan and organelle specific fluorescent probes revealed that Foscan presents low localisation in lysosomes and a weak accumulation in mitochondria. Alternatively, the Foscan fluorescence topographic profile turned out to colocalise perfectly with that obtained for the endoplasmic reticulum (ER) and the Golgi apparatus. The patterns of fluorescence derived from confocal microscopy studies were consistent with predominant localisation of Foscan in these organelles. Furthermore, evaluation of enzymatic activity of selected organelles immediately after laser light irradiation (650 nm) indicated the Golgi apparatus and ER as the primary damaged sites resulting from Foscan-mediated PDT in the MCF-7 cell line. To our knowledge, this is the first study to demonstrate unambiguously that the ER and the Golgi apparatus are preferential sites of Foscan accumulation. PMID- 12556975 TI - Alpha-tocopheryl succinate sensitises a T lymphoma cell line to TRAIL-induced apoptosis by suppressing NF-kappaB activation. AB - Activation of nuclear factor-kappaB (NF-kappaB) can interfere with induction of apoptosis triggered by the tumour necrosis factor-related apoptosis-inducing ligand (TRAIL; Apo2L). Therefore, agents that suppress NF-kappaB activation may sensitise cells to TRAIL-dependent apoptosis. Exposure of Jurkat cells to TRAIL resulted in massive and saturable apoptosis induction, following an initial lag time. This lag was abolished by pretreatment of the cells with subapoptotic doses of alpha-tocopheryl succinate (alpha-TOS) or the proteasome inhibitor MG132. Exposure of the cells to TRAIL led to a rapid, transient activation of NF-kappaB, a process that was suppressed by cell pretreatment with alpha-TOS or MG132. Activation of NF-kappaB by TNF-alpha prior to TRAIL exposure increased resistance of the cells to TRAIL-mediated apoptosis. We conclude that alpha-TOS sensitises cells to TRAIL killing, at least in some cases, through inhibition of NF-kappaB activation. This further supports the possibility that this semisynthetic analogue of vitamin E is a potential adjuvant in cancer treatment, such as in the case of TRAIL-mediated inhibition of cancer. PMID- 12556976 TI - Reply: Gallstones, cholecystectomy, and the risk for developing pancreatic cancer. PMID- 12556978 TI - [Experimental study on the biomechanical stability of different internal fixators for use in the mandible]. AB - BACKGROUND: Comparative studies of different forms of osteosynthesis require that biomechanical and biological criteria be observed. The conditions in experimental studies should be as close as possible to the in vivo situation. The aim of our study was to develop a fracture model that would allow determination of the micromovements in the gap tissue following different forms of internal fixation in fractured human mandibles. METHODS: Micromovements in the gap tissue of five human mandibles treated with different osteosynthetic systems (DCP, EDCP, Miniplates, 3-D systems) following osteotomy or fracture in the region of the corpus and median region were investigated by means of strain gauges. By fitting the human mandibles in plaster according to a method of our own it was possible to create fractures at predictable and comparable localizations. RESULTS: Our investigations show that the micromovements in the gap tissue of osteotomied and fractured mandibles are different and not dependent on the form of osteosynthesis applied. Physiological micromovements in the gap tissue were found under strain for all osteosynthetic systems used in fractured and congruently reset mandibles. DISCUSSION: Our fracture and osteosynthesis model allows the quantitative determination of micromovements in the gap tissue and shows the importance of ideal realignment, which has a decisive influence on micromovements in the gap tissue. PMID- 12556979 TI - [Distraction osteogenesis in the treatment of severe midfacial hypoplasia]. AB - PATIENTS: Between May 1998 and May 2002, 38 patients suffering from severe midfacial retrusion and atrophy were treated by way of midfacial distraction osteogenesis. Diagnoses included cleft lip and palate (32 patients) and one case of Crouzon's disease. Ages ranged from 6-65 years. A total of 28 patients presented a velopharyngeal flap and nine patients were almost or fully edentulous. Using an extraoral halo device, distraction was performed after a subtotal Le Fort-I/II/III or modified quadrangular osteotomy. RESULTS: Distraction ranged from 9 to 31 mm (17 mm average). Following the primary operation, seven patients underwent a second intervention due to problems with the procedure or the device. Two patients needed a secondary Le Fort-I-osteotomy. With respect to velopharyngeal insufficiency, 21% showed a deterioration and 8% an improvement. Postoperatively, a decrease of 15-20% in the attained sagittal advancement was seen during the first 6 months. This was attributed to relapses and postoperative orthodontics. Thereafter skeletal stability was maintained. CONCLUSION: Distraction osteogenesis of the midface can be the method of choice in severe midfacial retrusion. Due to the difficult patient situation and the technical intricacies a higher complication rate has to be accepted than for conventional dysgnathia operations. PMID- 12556980 TI - [Endoscopically assisted Le Fort 1 osteotomy: a study in human cadaver heads]. AB - METHODS AND RESULTS: Endoscopically assisted Le Fort 1-osteotomy was performed in a cadaver study with six human skulls. Vertical incisions were used to approach the maxilla using the endoscope. The bone cuts could be carried out with a 4-mm osteotome preserving the buccal mucosa. The downfracture of the maxilla was accomplished without damaging the descending palatal arteries, which could be verified endoscopically. Rigid fixation was performed using 1.5-mm titanium plates (Synthes, Mathys AG, Bettlach, Switzerland) and self-drilling screws. DISCUSSION: This cadaver study showed that the Le Fort 1 osteotomy could be carried out through limited approaches using endoscopic techniques and self drilling fixation tools. Further experimental work and improvements will help to introduce this technique into clinical application. PMID- 12556981 TI - [Post-traumatic ocular motility in orbital floor fractures]. AB - PURPOSE: The aim of this study was to evaluate post-traumatic ocular motility in orbital floor fractures in relation to the severity of the fracture and concomitant injuries. MATERIAL AND METHODS: Over a period of 6 years, data from 435 patients with orbital floor fractures were re-evaluated. Ocular motility was divided into abduction, adduction, upgaze and downgaze movements. RESULTS: In 54.3% of the cases, there was a reduction in ocular motility. A reduced motility with a movement >20 degrees was found in most of the cases. Concomitant injuries to the eye resulted in a significant increase in the reduction of motility. This reduction depended on the severity of the orbital floor fracture. A reduced upgaze movement was the most common complication in all types of fractures. CONCLUSION: Ocular motility can act as an indication of the severity of the orbital fracture. Concomitant injuries to the eye resulted in a higher incidence of reduced ocular motility. PMID- 12556982 TI - [Positioning of the premaxilla in cases of bilateral cleft lip, alveolus and palate using Latham's appliance]. AB - SUBJECT MATTER: In cases of bilateral cleft lip, alveolus and palate, preoperative positioning of the premaxilla using Latham's appliance has been described. This method is controversial, since it may cause growth defects. PATIENTS AND METHODS: From 1992 to 2000, Latham's appliance was inserted preoperatively into 20 patients with bilateral lip, alveolus and palate clefts with extreme premaxillary protrusion. Pin fixation ensued at the age of 3 months. After adjustment of the premaxilla, the soft palate was closed and bilateral lip adhesion as well as bilateral gingivoperiosteoplasty were performed directly after the removal of the appliance. Final lip closure took place 4-6 weeks later. At the age of 2 years, the hard palate was closed. Evaluation was based on combined face-maxilla models, standard photographs and, when available, lateral skull x-rays showing the relative position of the segments, the influence of Latham's appliance on the nasal septum and the relation of the upper jaw to the skull base and mandibula. RESULTS: In all cases, a satisfactory alveolar alignment was achieved; thickening and curvature of the nasal septum occurred but receded. Neither growth disturbances nor dental germ damage were seen. In two cases, complications arose from suture dehiscence of the lip adhesion: in one patient, a screw defect caused a loosening of the appliance and there was also a transmigration of the postpremaxillary pin. CONCLUSIONS: On the basis of our experience, the use of Latham's appliance, combined with consistent orthodontic supervision and, if necessary, treatment, represents a practical option for the treatment of bilateral cleft lip, alveolus and palate, especially in cases with extreme protrusion of the premaxilla. PMID- 12556983 TI - [Oxygen-carrying solutions improve tissue oxygenation in striated skin muscle subjected to critical ischemia]. AB - AIM: The aim of the present study was to investigate the effects of the oxygen carrying hemoglobin solution DCLHb (diaspirin-crosslinked hemoglobin) on microvascular perfusion and tissue oxygenation in striated skin muscle after the induction of critical ischemia followed by reperfusion. MATERIAL AND METHODS: Using intravital fluorescence microscopy the functional capillary density was analyzed in the striated skin muscle of Syrian golden hamsters before the induction of a 4-h period of ischemia and again after 0.5 h, 2 h and 24 h of reperfusion ( n=8 in each group). In other animals ( n=8 in each group), the identical protocol was applied to determine tissue oxygenation by means of the multi-wire surface electrode (MDO, Eschweiler, Kiel, Germany). Animals in the treatment group ( n=8) received a bolus infusion of 5 ml of DCLHb per kg of body wt. (10 g/dl; Baxter, Ill., USA) 15 min before reperfusion. Control animals ( n=8) received equivalent volumes of isotonic saline (Braun, Melsungen, Germany). RESULTS. Functional capillary density was dramatically reduced in control animals, while in DCLHb-treated animals significantly higher values were observed. Efficient restoration of tissue PO(2) was also seen in DCLHb-treated animals and not in control animals. CONCLUSIONS: These results show that the oxygen-carrying solution DCLHb is significantly more efficient than the commonly used crystalloid solutions in restoration of tissue PO(2) after ischemia reperfusion. The use of this solution therefore appears promising as a means of protecting the tissue put at risk by ischemia from reperfusion damage. PMID- 12556984 TI - [Involvement of the maxillary sinus in Langerhans cell histiocytosis]. AB - AIM: A clinical presentation of Langerhans cell histiocytosis (LCH) in the maxillary sinus of two patients is given. LCH in the maxillary sinus is a rare occurrence. Our aim was to compare the different treatment alternatives available and to suggest a classification as well as a therapeutic regime. PATIENTS AND METHOD: Records and clinical data of two patients treated between 1994 and 2001 were retrospectively evaluated. Both patients suffered from LCH in the maxillary sinus and the maxilla regions. Only surgical treatment was used. After resection, a large defect of the maxillary sinus, which did not allowing coverage, was seen in both cases. After reconstructive operations, closure was finally achieved. Both patients underwent follow-ups, whereby one suffered from a relapse after 15 months. RESULTS: Although one of the patients under investigation showed a recurrence of LCH, we are of the opinion that surgical treatment is very effective in the elimination of this condition. A proposal for a classification of LCH in the oral-maxillo-facial-region is made. CONCLUSIONS: The evaluation of our clinical study suggests that LCH is a disease that should be treated surgically. Only in very severe cases should the surgical treatment be complimented by either radiotherapy or chemotherapy. In disseminated cases, especially chemotherapy seems to improve the outcome. Surgery offers the possibility of eliminating systemic side effects. PMID- 12556985 TI - [Criteria of success in implantology]. AB - BACKGROUND: So far there is no generally accepted definition of success in the case of dental implants, although criteria differing from country to country have been proposed, which evaluate the success of implants with reference to clinical and X-ray control parameters. The aim of the study was to examine the five most frequently used sets of criteria suggested for determination of the success or otherwise of implants ( Albrektsson, Jahn, Buser, NIH conference, Naert) and ascertain their relative values. METHODS: In a clinical review, 508 implants (Astra, Frialit-2, IMZ TwinPlus and ITI) were examined and a subjective evaluation of them was made by the patients. Subsequently a statistical analysis was executed (in situ rate, Kaplan-Meier, success criteria). RESULTS: The average observation time of all implants included in the study was 3.5 years, and the in situ survival rate was 95%; Kaplan-Meier survival analysis revealed that the probability of survival after 6 years was 93%. Application of different criteria for success gave very different results (Albrektsson 88%, Naert 89%, NIH 85%, Buser 88%, Jahn and dacute;Hoedt 75%). DISCUSSION: The increasing importance of quality assurance in medicine makes it necessary to establish standards that will permit reliable comparative analysis of how successful implants are in the future. PMID- 12556986 TI - [Juvenile ossifying fibroma. Case report with diagnostic and therapeutic considerations]. AB - BACKGROUND: Fibro-osseous lesions of the jaws, including juvenile ossifying fibroma, pose diagnostic and therapeutic difficulties due to their clinical, radiological and histological variability. CASE REPORT: The case of an 8-year-old girl with a juvenile ossifying fibroma of the mandible is presented. Extirpation of the tumor and filling of the defect with cancellous bone was performed. After 9 months, a recurrence occurred which made a second extirpation of the tumor necessary. This was done without osteoplasty. After a 2-year follow-up the patient is free of tumor. DISCUSSION: Different classifications for fibro-osseous lesions of the jaws can be found in the literature. This makes a correct diagnosis and standardized treatment difficult. Based on the case presented here, the histological features of juvenile ossifying fibroma and the differentiation from osteosarcoma are discussed. At present, conservative surgical treatment seems to be adequate for these tumors. PMID- 12556987 TI - [Verruciform xanthoma]. AB - BACKGROUND: Verruciform xanthoma (VX), a rare, benign lesion of the skin and mucosa, is slow-growing, asymptomatic and characterized by a granular (verruciform) surface. It is yellowish-red or grey in color and up to 2 cm in diameter. Histologically, a papillary and/or verrucous proliferation of the squamous epithelium with hyperparakeratosis and numerous foam cells is present. These cells are predominantly located within the papillae of the lamina propria. For differential diagnosis, other papillomatous and verrucous lesions such as verrucous carcinomas or squamous cell carcinomas need to be ruled out. CASE REPORT: A 46-year-old patient with VX located on the alveolar process regio 26-28 is presented. Clinically, a 2 x 2 cm granular, oral mucosa surface lesion extending onto the palate occurred in regio 26-28. Biopsy was characterized light microscopically by the presence of swollen, elongated cells in the submucosa, an indication of VX alterations. Transmission electron microscopy demonstrated foam cells in the subepithelium containing numerous membrane-bound vesicles similar in diameter and showing a wide variation in electron density. Morphologically, these cells resembled macrophage-related cells. The lesion was excised in total with no evidence of recurrence after 9 months. DISCUSSION: The pathogenesis of VX is still unclear. The characteristic xanthoma cells may play a major role in VX. Microscopic analysis of the morphology of the foam cells indicated that they may represent a differentiated form of macrophages. Lipid vesicles inside these cells differed in their electron density indicating a heterogeneous biochemistry or different states of maturation. PMID- 12556988 TI - Comparison of the amino acid sequences of RNA-dependent RNA polymerases of cypoviruses in the family Reoviridae. AB - The nucleotide sequences of the genome segment S2 of Bombyx mori cypovirus 1, S2 of Lymantria dispar cypovirus 1, S1 of Lymantria dispar cypovirus 14 and S1 of a proposed new electropherotype of Trichoplusia ni cypovirus 15 were determined. These segments encoded putative RNA-dependent RNA polymerases (RDRPs). The deduced amino acid sequences of RDRPs within the genus Cypovirus showed 32% to 94% identities, while extent of homology between RDRPs in the genera Cypovirus and Oryzavirus, a genus most closely related, was approximately 26% identity. Both the genera Cypovirus and Oryzavirus might have originated from a common insect virus ancestor. PMID- 12556989 TI - Effect of Ca2+-dependent cell death on the release of herpes simplex virus. AB - The effect of a variety of cell death-inducing reagents on the release of herpes simplex virus type 1 (HSV-1) was examined. Ionomycin was found to increase the release of HSV-1, whereas no significant increase was observed by the treatment with TNF-alpha, anti-Fas antibody, C2-ceramide, sphingosine, H-7, tyrphostin and camptothecin. Ionomycin induced an immediate early peak and a subsequent long lasting elevation of intracellular Ca(2+) concentration ([Ca(2+)]i). In the absence of extracellular Ca(2+), ionomycin neither elevated [Ca(2+)]i nor increased the release of HSV-1 from the infected cells, indicating that Ca(2+) influx play an important role in the release of HSV-1. Studies with trypan blue and annexin V staining revealed that the ionomycin-induced alteration of [Ca(2+)]i was accompanied by cell death of the infected cells. Disintegration of cell membrane, cytoplasmic vacuole formation and the leakage of virus particles from the cell surface were observed by electron microscopy. These results indicate that Ca(2+)-dependent cell death showing necrotic alteration is responsible for the enhanced release of HSV-1. The data also give some initial insights into the functional importance of cell death during the late stages of HSV-1 infection. PMID- 12556990 TI - Suppression of the ERK1/2 signaling pathway from HCV NS5A protein expressed by herpes simplex recombinant viruses. AB - Two herpes simplex type 1 (HSV-1) recombinant viruses carrying the hepatitis C virus (HCV) NS5A open reading frame under the control of the cytomegalovirus immediate early (IE) or a herpes simplex chimeric promoter (alpha 4 gamma 1UL19) were constructed and characterized. Expression studies showed that both HSV-NS5A recombinant viruses were able to express high levels of the NS5A protein in infected cells. Most importantly, using this system, we demonstrated that the NS5A protein interacts with the growth receptor-bound protein 2 (Grb2) and inhibits the phosphorylation of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) in HeLa, NIH3T3 or liver infected cells. Thus, our studies confirm the ability of the NS5A protein to perturb the extracellular signal-regulated kinase (ERK) pathway in HeLa cells by the use of an alternative system for NS5A expression and extend this observation to additional cell lines. We conclude that HSV-based viral vectors may provide a useful system for studying the expression and selected functional properties of the HCV NS5A protein. PMID- 12556991 TI - Genetic diversity of enterovirus 71 isolated from cases of hand, foot and mouth disease in Yokohama City between 1982 and 2000. AB - Enterovirus 71 (EV71) is known as one of the major causative agents of hand, foot and mouse disease (HFMD) and is also associated with neurological manifestations such as aseptic meningitis, polio-like paralysis and encephalitis. Recently, large HFMD outbreaks, involving severe neurological complications, have been experienced in Malaysia, Taiwan and some other countries in the Western-Pacific region. To investigate the genetic diversity of EV71 isolates in a single community in Japan, nucleotide sequences of the VP4 region of 52 EV71 isolates in Yokohama City from 1982 to 2000 were determined and the phylogenetic relationship was compared with other referential EV71 strains in Japan and in the world. There were two major genotypes of EV71 in Yokohama City through the 1980's and 1990's. Six EV71 isolates in the early 1980's in Yokohama City were closely related to those from HFMD outbreaks in Japan and from outbreaks of polio-like paralysis in Europe in the 1970's. During recent HFMD outbreaks in 1997 and 2000, two distinct genotypes of EV71 were co-circulating in Yokohama City as in HFMD outbreaks in Malaysia and Taiwan. However, the genetic diversity of EV71 in Yokohama City was not directly correlated with the severity of HFMD. The results confirmed the circulation of two distinct genotypes of EV71 over the past 20 years in Japan. PMID- 12556992 TI - Studies on the C-terminus of the Cowpea mosaic virus movement protein. AB - Cowpea mosaic virus (CPMV) spreads from cell-to-cell as virus particles through tubular structures in modified plasmodesmata which are composed of viral movement protein (MP). Mutational analysis of the MP has revealed that the N-terminal and central regions of the MP are involved in tubule formation and that the C terminal domain probably has a role in the interactions with virus particles. By constructing C-terminal deletion mutants and comoviral hybrid MPs, it was possible to delineate the C-terminal border of the tubule-forming domain to a small region between amino acids 292 and 298. Experiments with tripartite viruses in protoplasts indicated that the C-terminus of the MP is involved in the incorporation of virus particles in the tubule and that for efficient incorporation of virus particles all MP molecules incorporated in a tubule need to contain a functional C-terminus. A mutant virus coding for a MP in which the last 10 C-terminal amino acids were replaced by the green fluorescent protein (GFP) was able to form tubules in protoplasts. These tubules did not contain virus particles, probably because the GFP interferes with the incorporation of virions into the tubule. These results suggest a model for the structure of the tubule in which the C-terminus of the MP is located inside the tubular structure, where it is able to interact with virus particles. PMID- 12556993 TI - Distribution and genetic diversity of tomato-infecting begomoviruses in Brazil. AB - Tomato-infecting begomoviruses have been reported throughout Brazil since the introduction of the B biotype of Bemisia tabaci. Here, we report a large scale survey on the distribution and genetic diversity of tomato-infecting begomoviruses. Tomato samples with typical begomovirus symptoms were collected in seven different states, comprising the major tomato growing areas of the country. Viruses were detected by polymerase chain reaction (PCR) using universal primers for the genus Begomovirus. PCR-amplified fragments were cloned and sequenced. Based on sequence comparisons and phylogenetic analyses, at least seven previously undescribed species of begomoviruses were found. Four of the new viruses were found exclusively in the Southeastern states, two exclusively in the Northeastern states, and one was found in both regions. Sequence comparisons reveal strong evidence of recombination among the Brazilian begomoviruses. Together, the results indicate the existence of a high degree of pre-existing genetic diversity among tomato-infecting begomoviruses in Brazil and suggest that these viruses have emerged after being transferred from natural hosts to tomatoes, due to the introduction into Brazil of a novel polyfagous biotype of the whitefly vector. PMID- 12556994 TI - Pathogenicity and kinetics of virus propagation in swine infected with the cytopathogenic classical swine fever virus containing defective interfering particles. AB - To analyze the pathogenicity and in vivo kinetics of the cytopathogenic (cp) classical swine fever virus (CSFV) WB82 strain, which is composed of cp defective interfering (DI) particles and noncytopathogenic (noncp) helper virus (WB82/E(+) strain), WB82 and WB82/E(+) strains were administered separately to domestic pigs. After inoculation with either strain, all pigs showed typical symptoms of classical swine fever (CSF), such as leucopenia and high fever. There were few differences in clinical signs and survival times between each group. However, the appearance of some symptoms of CSF had a tendency to be delayed following infection with the WB82 strain, when compared with the WB82/E(+) strain. Virus isolation and detection of subgenomic (sg) and full-length viral (flv) RNA by RT PCR was carried out using sera, 10% homogenized organs and oral, nasal and rectal swabs. Both noncytopathogenic helper virus and cp DI particles were detected in samples from pigs infected with the WB82 strain, but only noncp phenotype virus was isolated from pigs infected with the WB82/E(+) strain. Interestingly, the cp DI particles appeared six to seven days later than helper virus in sera from pigs infected with the WB82 strain. Although active cp DI particles could not be isolated from swabs, sg RNA as well as flv RNA was detected in swabs from animals infected with the WB82 strain. These results suggest that progeny cp DI particles are replicated from parent DI particles after noncp virus replication, and subsequently discharged from infected animals. Furthermore, propagation of DI particles or replication of sg RNA, following propagation of helper virus, appears to inhibit the appearance of CSF symptoms induced by virulent helper CSFV. PMID- 12556995 TI - Construction and in vivo detection of an enhanced green fluorescent protein expressing strain of Saimiriine herpesvirus 1 (SaHV-1). AB - Saimiriine herpesvirus 1 (SaHV-1) is an alpha-herpesvirus of squirrel monkeys used in mice to study neural pathogenesis of herpesviruses. To trace dissemination of virus from a peripheral site of inoculation to the central nervous system tissues, a recombinant strain of SaHV-1 expressing enhanced green fluorescent protein (GFP) was constructed by site-specific insertion of a GFP expression cassette into a transcriptionally null point in the SaHV-1 genome. PCR and Southern blot confirmed insertion of a single GFP expression cassette into the target site of the SaHV-1 genome. The recombinant virus was shown to produce strong fluorescence in the cytoplasm of infected cells in vitro. Growth kinetic experiments demonstrated no differences between recombinant and wild type SaHV-1 in producing infectious progeny virions. The recombinant virus was comparable to wild type SaHV-1 in development of clinical disease, microscopic lesions and induction of an antibody response in mice following intramuscular inoculation. Using confocal microscopy, GFP expression was easily observed in formalin fixed, paraffin-embedded tissues of mice infected with the recombinant SaHV-1. This simple specimen processing technique preserves tissue morphology and allows detection of viral replication within various tissues of experimentally infected animals. PMID- 12556996 TI - Multiple mechanisms for HSV-1 induction of interferon alpha production by peripheral blood mononuclear cells. AB - UV-inactivated, infectious, and other forms of herpes simplex virus 1 (HSV-1) induced interferon (IFN) production by different routes in myeloid origin mononuclear cells (MOMC) (consisting predominantly of monocytes). GM-CSF activated the MOMC (G-MOMC) to produce greater amounts of interferon while differentiation to DC, by the addition of granulocyte macrophage colony stimulating factor (GM-CSF) and calcium ionophore (GA-MOMC), reduced the levels of interferon production upon challenge with some HSV strains. UV-inactivated virus induced more interferon than infectious virus. L-fucose, an antagonist of the mannose receptor, inhibited the induction of IFN-alpha by UV-inactivated virus and gB(-) virus (defective in penetration) in MOMC and GA-MOMC but not G MOMC. L-fucose had little effect on interferon induction by infectious HSV-1. The insensitivity of the G-MOMC to fucose inhibition distinguishes these interferon producing cells from the pDC2 cells previously described as natural interferon producing cells. The mannose receptor appears to be involved in the response to non-infectious forms of HSV but infectious virus appears to use a different pathway. These studies suggest that non-infectious virions and HSV infected cell debris effectively stimulate monocytes and pre-dendritic cells to produce IFN alpha to initiate host protection against HSV infection. PMID- 12556997 TI - Involvement of the terminus of grouper betanodavirus capsid protein in virus-like particle assembly. AB - Dragon grouper, Epinephelus lanceolatus, nervous necrosis virus (DGNNV) consists of 180 copies of capsid protein that encapsulates a bipartite genome of single stranded (+)RNAs. Expressing the open reading frame (ORF) of RNA2 in Escherichia coli forms virus-like particles (VLPs) that resembles native virus. Deleting N- and C-termini revealed different impacts on VLP formation. Deletion of 35 or 52 residues at the N-terminus completely ruined the VLP assembly, presumably due to removal of positively charged residues for binding RNAs. When deletions were restricted to 4, 16, or 25 N-terminal residues, the assembly of VLPs remained. The ability of VLP formation diminished when 4 to 11 C-terminal residues were deleted. The termini that can be deleted without seriously destructing the VLPs are 25 and 3 residues at N- and C-termini, respectively. PMID- 12556998 TI - Genetic diversity in the coat protein coding region of eighty-six sugarcane mosaic virus isolates from eight countries, particularly from Cameroon and Congo. AB - Fifty-eight sugarcane virus isolates were obtained from leaves showing mosaic symptoms, and collected in Cameroon (26 isolates), Congo (20 isolates), Egypt (1 isolate), South Africa (3 isolates) and the U.S.A. (8 isolates). All these isolates belonged to Sugarcane mosaic virus (SCMV) based on the amplification product obtained by RT-PCR with SCMV-specific primers. The amplicons (0.9 kb) from the coat protein (CP) coding region were cloned, sequenced and compared to each other as well as to the sequences (GenBank accessions) of 16 SCMV isolates from sugarcane (Australia, South Africa and U.S.A.) and 12 SCMV isolates from maize (Australia, Germany and China). Maximum likelihood and maximum parsimony analyses robustly supported two major monophyletic groups that were correlated with the host of origin: the SCE or sugarcane group that included all isolates from sugarcane and the MZ or maize group that contained all isolates from maize. The 86 virus isolates were distributed in 13 minor phylogenetic groups, four (I IV) restricted to maize and nine (V-XIII) to sugarcane. A strong correlation was observed between the sugarcane groups and the geographical origin of the SCMV isolates. Each SCMV type strain from sugarcane (A, B, D, E and SC) was distributed in a different phylogenetic group or subgroup. The 26 isolates from Cameroon constituted a relatively homogeneous group (group V) whereas the 20 isolates from Congo belonged to two other groups (VI and VII). All the isolates from Cameroon and Congo were different from the SCMV type strains and other strains or isolates studied so far. It appears, therefore, that the population of SCMV from sugarcane in Africa contains virus genotypes that have not yet been described. PMID- 12556999 TI - Genomic sequence of Wild potato mosaic virus as compared to the genomes of other potyviruses. AB - The complete nucleotide sequence of Wild potato mosaic virus (WPMV) was determined, showing that it is a distinct member of the genus Potyvirus (family Potyviridae). The genome consists of 9853 nucleotides and encodes a single polyprotein of 3065 amino acids. The 5'- and 3'-non translated regions (NTR) were 182 and 472 nucleotides, respectively. Alignment of the predicted WPMV polyprotein sequence with other members of the genus Potyvirus revealed nine putative cleavage sites resulting in ten mature proteins. Amino acid and nucleotide sequence identities and the exceptionally long 3'-NTR indicated a close relationship between WPMV and Potato virus V (PVV). Phylogenetic analysis based on the polyprotein and coat protein amino acid sequences grouped WPMV, PVV, Pepper mottle virus and Potato virus Y in one cluster distinct from other potyviruses. PMID- 12557000 TI - Identification of two nucleotide sequence sub-groups within Potato mop-top virus. AB - To evaluate the variation of Potato mop-top pomovirus from potato fields, 21 isolates were collected from different Danish locations. Reverse transcription polymerase chain reaction-restriction fragment length polymorphism (RT-PCR-RFLP) of regions of RNA2 was performed for all 21 isolates resulting in the establishment of two sub-groups of isolates. The nucleotide sequence of a region encoding part of the 'readthrough protein' of RNA2 was compared for 9 of these isolates. This sequence analysis confirmed the RT-PCR-RFLP grouping. The isolates were tested for symptom expression in indicator plants and grouped according to symptom development. No correlation between grouping based on symptom development and genotype was observed. PMID- 12557001 TI - Complete nucleotide sequence and genome organization of a Chinese isolate of tobacco bushy top virus. AB - The complete nucleotide sequence of a Chinese isolate of tobacco bushy top virus (TBTV), designated TBTV-Ch, was determined from cDNA generated from double stranded RNA extracted from diseased tobacco. The genome is 4152 nucleotides (nt) in size, contains four putative open reading frames (ORFs) and untranslated regions of 10 nt and 645 nt at the 5' and 3' ends, respectively. In genome organization and in the amino acid sequence of its potential products, the RNA of TBTV-Ch is similar to other umbraviruses sequenced to date. The results suggested that TBTV should be regarded as a definitive species of the genus Umbravirus. PMID- 12557002 TI - First detection of group C rotavirus in children with acute diarrhea in Spain. AB - Group C rotavirus causes sporadic cases and outbreaks of acute diarrhea in children and adults in many countries, but has never been detected among children in Spain. In a recently conducted surveillance study to screen fecal specimens for bacteria and viruses from a cohort of 822 young children who were treated for acute diarrhea in Madrid, no pathogens were detected in fecal specimens from 238 (29%) children. In this study, we examined 147 of those specimens for group C rotavirus by EIA and PCR and found 22 (15%) were positive. Our findings demonstrate that group C rotavirus is an important cause of childhood diarrhea in Spain. PMID- 12557003 TI - Revision of taxonomic criteria for species demarcation in the family Geminiviridae, and an updated list of begomovirus species. PMID- 12557004 TI - Postischemic neuroprotection in the ischemia-tolerant state gerbil hippocampus is associated with increased ligand binding to inhibitory GABA(A) receptors. AB - Excitotoxic activation of glutamate receptors is thought to play a key role in delayed neuronal death (DND) of highly vulnerable hippocampal CA1 neurons after transient global ischemia. DND can be prevented by a short sublethal preconditioning (PC) stimulus. Recently, we demonstrated that ischemic PC, but not a single period of 5-min ischemia elicits a transient up-regulation of hippocampal [(3)H]muscimol binding to GABA(A) receptors. This indicates that activation of the GABAergic system may participate in the acquisition of neuroprotection. The present study was designed to test whether postischemic modulation of receptor binding also occurs in the ischemia-tolerant state, i.e., after a PC stimulus of 2.5-min ischemia and a subsequent normally lethal period of 5-min ischemia 4 days apart. Using receptor autoradiography, [(3)H]AMPA and [(3)H]muscimol binding to excitatory AMPA and inhibitory GABA(A) receptors was analyzed in hippocampal subfields CA1, CA3 and dentate gyrus at recirculation intervals of 30 min, 8, 24, 48, 96 h and 3 weeks. Postischemic hippocampal ligand binding to AMPA receptors remained unchanged at any time point investigated, but [(3)H]muscimol binding to GABA(A) receptors in CA1 neurons rendered tolerant to ischemia was up-regulated between 30 min and 48 h of recirculation. Our data suggest that a relative shift between excitatory and inhibitory neurotransmission may promote postischemic survival of CA1 neurons. PMID- 12557005 TI - Oct6, a transcription factor controlling myelination, is a marker for active nerve regeneration in peripheral neuropathies. AB - Three transcription factors, Krox20 (EGR2), Oct6 (SCIP/Tst1) and Sox10, are considered necessary for transition from the nonmyelinating to the myelinating stage of Schwann cell development. We immunohistochemically studied Oct6 expression in peripheral nerve specimens from 25 patients with various diseases including Charcot-Marie-Tooth disease type 1A (CMT1A). Oct6 was present in cytoplasm of Schwann cells associated with normal-appearing myelinated nerve fibers, but not in nuclei. Expression was seen in nuclei of Schwann cells in the early phase of acute axonal degeneration; nuclear expression peaked at the regenerative stage. Schwann cells forming "onion bulbs" expressed Oct6 in chronic inflammatory demyelinating polyneuropathy (CIDP), but showed minimal expression in CMT1A, reflecting their proliferative activity in CIDP. Nerves showing chronic axonal loss had no expression. Oct6, then, may be a marker for dedifferentiation of adult Schwann cells and active nerve regeneration. PMID- 12557006 TI - Cytokine immunoreactivity in cortical and subcortical neurons in periventricular leukomalacia: are cytokines implicated in neuronal dysfunction in cerebral palsy? AB - The major neuropathological substrate associated with cerebral palsy (CP) is a form of white matter (WM) injury known as periventricular leukomalacia (PVL). Proinflammatory cytokines were recently shown to be implicated in PVL pathogenesis. Many PVL patients develop cortical and deep gray neuronal dysfunctions such as epilepsy, cognitive deficits and extrapyramidal disorders. The precise nature of the relationship between the WM lesion and the subsequent neuronal disorders is unclear. Cytokines were shown to exert neurotoxicity in experimental models. This raises the need to investigate a possible noxious effect by cytokines on neuronal cortical development. In situ immunohistochemical methods were applied on 22 brains from infants both with PVL (study group) and without PVL (control group) to detect any immunoreactivity for cytokines (TNF alpha, IL-1beta, IL-6) in cortical and gray matter neurons. While cortical and other neuronal structures in PVL brains did not display noticeable pathological anomalies, strong cytokine immunoreactivity was detected in many neurons in the neocortex, hippocampus, basal ganglia and thalamus. There were, however, regional differences in cytokine labeling. In addition, there was more TNF-alpha staining than IL-1beta; IL-6 was negative. In contrast, neuronal cytokine labeling in the "control" brains was negligible. In conclusion, we report and characterize, for the first time, the in situ immunoreactivity for proinflammatory cytokines in cortical and deep gray neurons in PVL. These findings might provide insights into the neuro-anatomical correlate for the intellectual deficits and the other cortical and deep gray neuronal dysfunctions associated with PVL. PMID- 12557007 TI - Expression of tumor necrosis factor-alpha in regenerating muscle fibers in inflammatory and non-inflammatory myopathies. AB - The expression level of tumor necrosis factor (TNF)-alpha is elevated in idiopathic inflammatory myopathies and Duchenne muscular dystrophy (DMD), but the precise role of TNF-alpha is unknown. To elucidate the possible role of TNF alpha, we investigated the expression of TNF-alpha and its receptor in polymyositis (PM), dermatomyositis (DM), and DMD using in situ hybridization (ISH) and immunohistochemistry. We showed that TNF-alpha mRNA and protein were present in muscle fibers. TNF-alpha-positive fibers were observed in all cases of PM, DM and DMD, but were rare or absent in neurogenic disorders and normal controls. The proportion of TNF-alpha-positive fiber showed a significant positive correlation with the proportion of regenerating fibers that were positive for the developmental form of myosin heavy chain (MHC-d). The number of TNF receptor-positive fibers was small. Some muscle fibers expressed both TNF alpha and its receptor simultaneously. Our results indicate that TNF-alpha is produced and expressed by muscle fibers and associated with muscle regeneration. PMID- 12557009 TI - Apoptosis and microglial activation in influenza encephalopathy. AB - During influenza epidemics in Japan, the number of children with acute encephalopathies and encephalitis has recently increased. Although the pathophysiologies remain unclear, there is usually brain edema with evidence of damage to the blood-brain-barrier (BBB). We investigated the glial reaction and apoptosis in brains of eight such cases comprising two of acute necrotizing encephalopathy and six of influenza encephalopathy, and compared the results with those in five control brains. Apoptosis, evidenced by chromatin condensation and fragmentation in hematoxylin sections, in situ end labeling of fragmented DNA (TUNEL) and DNA laddering, was observed in neurons and glial cells in four brains with influenza encephalopathy. In the TUNEL-positive brains, the increase in microglia was greater than in the TUNEL-negative brains. Immunoreactivity for active-caspase 3, demonstrated by immunohistochemistry, and the overexpression of a caspase-cleaved fragment of poly(ADP-ribose) polymerase, demonstrated by Western blotting, indicated that activation of caspase 3 is involved in the apoptotic pathway in the brains of influenza encephalopathy cases. Apoptosis or specific pathological processes that cause apoptosis may give rise to aggravated encephalopathy. PMID- 12557008 TI - The dentate gyrus neurogenesis: a therapeutic target for Alzheimer's disease. AB - Neurogenesis persists in the aged human dentate gyrus but its role and regulation in pathological conditions such as Alzheimer's disease (AD), where the neurotrophic environment is changed, are poorly understood. In this study we investigated the effect of changes in the neurotrophic environment on neurogenesis in cultured rat hippocampal progenitors and in normal adult rats as models. In hippocampal progenitor cells from adult rats, fibroblast growth factor 2 (FGF-2) dose-dependently decreased microtubule-associated protein 2 and increased tau levels, indicating an FGF-2-induced dendrite to axon polarity shift. Cerebrolysin, a neurotrophic drug which has been shown to improve cognition and mood of AD patients, was found to increase neuron-like differentiated adult rat hippocampal progenitors in culture both by reducing apoptosis and by counteracting the FGF-2-induced polarity shift. Intraperitoneal administration of Cerebrolysin enhanced dentate gyrus neurogenesis and maze performance of 8- to 12-month-old female rats. These studies suggest that AD pathogenesis might involve an abnormally elevated FGF-2-associated dysregulation of dentate gyrus neurogenesis, especially neuronal polarity and that the neurogenesis pathology is a promising therapeutic target for this disease. PMID- 12557010 TI - Meningial blood vessel calcification in the brain of the cat. AB - Mineralization in the wall of central nervous system blood vessels is sporadically encountered in aged horses and cattle as in man, generally as an age related change. This phenomenon has not to date been located in the meninges in dogs or cats. The present study reports a retrospective histological examination of 50 feline brains from 40-day- to 13-year-old cats. Histological examination using routine staining techniques (hematoxylin and eosin, Luxol fast blue periodic acid-Schiff) and special stains (Von Kossa and Pearl's method) showed substantial blood vessel calcification (BVC) in 29 cases which, except for 1 case, was present only in the leptomeninges. In 72% of cases BVC was not related to nervous tissue lesions. For this reason it was considered an incidental finding, producing no morphological or clinical signs. However, BVC should not be considered merely an age-related finding since it is also quite common in very young animals (35%), suggesting that its pathogenesis needs to be investigated further and compared to BVC observed in children affected by acquired immune deficiency and idiopathic arterial calcification. PMID- 12557011 TI - Severe depletion of mitochondrial DNA in spinal muscular atrophy. AB - Spinal muscular atrophy (SMA) is a neuromuscular disorder in childhood leading to a dramatic loss of muscle strength. Functional investigations with high resolution polarography and enzyme measurements of the respiratory chain revealed lowered activities in muscle tissue of SMA patients. To gain a better understanding of this low energy supply we analyzed the amount of mitochondrial DNA (mtDNA) in skeletal muscle of 20 unrelated children with genetically proven SMA and 31 controls. Quantitative Southern blot analysis revealed a severe and homogeneous decrease in the content of muscle mtDNA in relation to nuclear DNA in SMA patients (90.3+/-7.8%), whereas by immunofluorescence no decrease in the number of mitochondria was detected. In addition, a two- to threefold reduction of the nuclear-encoded complex II (succinate dehydrogenase) activity was detected in SMA muscle tissue. Western blot analysis showed a significant reduction of both mitochondrial- and nuclear-encoded cytochrome c oxidase subunits. Our results indicate that mtDNA depletion in SMA is a consequence of severe atrophy, and has to be differentiated by measurement of complex II from an isolated reduction of mtDNA as found in patients with mitochondriocytopathies and the so called mtDNA depletion syndrome. PMID- 12557012 TI - Abeta species, including IsoAsp23 Abeta, in Iowa-type familial cerebral amyloid angiopathy. AB - Mutations within the Abeta sequence of the AbetaPP gene are associated with familial forms of cerebral amyloid angiopathy (CAA). One mutation, Abeta D23N, was identified in a family in Iowa with a clinical history of early-onset dementia. We analyzed the pattern of Abeta deposition in the hippocampus of an individual with Iowa CAA. In addition to strong amyloid angiopathy, we found unusual diffuse Abeta deposits in the CA4, and in the parenchyma near amyloid laden vessels. ELISA of cortical brain extracts showed that Abeta40 was nearly 20 fold higher than Abeta42, in both soluble and insoluble fractions. We identified an Abeta antibody that recognized wild-type Abeta but not Iowa Abeta. With this antibody, we found that wild-type Abeta was present in the Abeta deposits, but limited to the strongest deposits in the cerebrovasculature. Previous in vitro studies suggested that the presence of an asparagine at position 23 of Abeta favored formation of an isoAsp residue, which was associated with increased Abeta fibrillogenesis. Using isoAsp-specific antibodies in immunohistochemical studies, we examined the distribution of isoAsp Abeta in the Iowa brain. IsoAsp7 Abeta was present in both the parenchymal and vascular deposits, whereas isoAsp23 Abeta was present only in vascular deposits. These data suggest that alteration of Abeta Asn23 to isoAsp may be an important determinant in the deposition of Abeta in cerebral blood vessels. PMID- 12557013 TI - Immunohistochemical examination on intracranial calcification in neurodegenerative diseases. AB - Fahr-type calcification is a relatively common finding in the elderly, and in younger patients with Alzheimer's disease, calcification in the basal ganglia is not uncommon. However, as far as we know, an immunohistochemical study of intracranial calcification in neurodegenerative diseases has not been performed. In this study, we examined intracranial calcification of the basal ganglia and cerebellum with antibodies against noncollagenous bone matrix proteins. Nineteen brains were employed. The diagnoses were diffuse neurofibrillary tangles with calcification in five, Alzheimer's disease in five, Pick's disease in one, progressive supranuclear palsy in one, Parkinson's disease in one, and six controls. By conventional histology, three patterns of calcium (Ca) deposition were recognized: diffuse deposition within the tunica media of small and medium sized vessels (type 1 deposition), free spherical or lobulated concretions (type 2 deposition) in the parenchyma, and rows of small calcospherites lying along capillaries (type 3 deposition). Type 3 deposition is relatively rare, and may be a hallmark of severe intracranial calcification. Immunohistochemistry demonstrated that osteopontin was present diffusely in all Ca deposition types. Osteocalcin was present chiefly in the peripheral region of type 2 and 3 depositions, as well as in only the rims of type 1 deposition. Bone sialoprotein and osteonectin were found only in the core portions of type 2 and 3 depositions. In brief, type 1 deposition shows a different staining pattern from type 2 and 3. Different Ca deposition patterns of noncollagenous bone matrix proteins may suggest their separate roles in the pathogenesis of intracranial calcification. PMID- 12557014 TI - Dementia with Lewy bodies from the perspective of tauopathy. AB - We immunohistochemically investigated the prevalence and pattern of phosphorylated tau accumulation in neurons and glia in 46 cases of dementia with Lewy bodies (DLB). Tau-positive neurons composed of neurofibrillary tangles (NFT) and pretangle neurons were found in the hippocampal area in all 46 cases, although the ratio of pretangle neurons in tau-positive neurons was higher in the cases showing low NFT stages. Tau-positive astrocytes were found in the periventricular area in 18 of 46 cases, and partly represented argyrophilic thorn shaped astrocytes. In contrast, tau-positive oligodendroglia were found in the subcortical white matter in 9 of 46 cases, and represented argyrophilic coiled bodies. Tau-positive argyrophilic grains were found in the hippocampal area in the same cases as those with coiled bodies. The 9 cases with tau-positive coiled bodies and grains were included in the 18 cases with tau-positive astrocytes, and showed larger proportions in the low NFT stages than the 46 cases with tau positive neurons. Tau-positive neurons were positive both to anti-three-repeat (3R) and -4R tau-specific antibodies, while tau-positive astrocytes, coiled bodies and grains were predominantly positive to anti-4R tau-specific antibody. These tau-positive structures were negative to anti-alpha-synuclein antibody. These findings suggest that the tau accumulation in DLB represents both tau positive neurons with all six tau isoforms and tau-positive astrocytes, coiled bodies and grains with the 4R tau isoform, and that the different cytoskeletal abnormalities form a link between some neurodegenerative dementing disorders including DLB. PMID- 12557015 TI - Calcineurin is a potent regulator for skeletal muscle regeneration by association with NFATc1 and GATA-2. AB - The molecular signaling pathways involved in regeneration after muscle damage have not been identified. In the present study, we tested the hypothesis that calcineurin, a calcium-regulated phosphatase recently implicated in the signaling of fiber-type conversion and muscle hypertrophy, is required to induce skeletal muscle remodeling. The amount of calcineurin and dephosphorylated nuclear factor of activated T cells c1 (NFATc1) proteins was markedly increased in the regenerating muscle of rats. The amount of calcineurin co-precipitating with NFATc1 and GATA-2, and NFATc1 co-precipitating with GATA-2 gradually increased in the tibialis anterior muscle after bupivacaine injection. Calcineurin protein was present in the proliferating satellite cells labeled with BrdU in the damaged muscle after 4 days. In contrast, calcineurin was not detected in the quiescent nonactivating satellite cells expressing Myf-5. At 4 days post injection, many macrophages detected in the damaged and regenerating area did not possess calcineurin protein. Calcineurin protein was abundant in many myoblasts and myotubes that expressed MyoD and myogenin at 4 and 6 days post injection. In the intact muscle, no immunoreactivity of calcineurin or BrdU was detected in the cell membrane, cytosol or the extracellular connective tissue. In mice, intraperitoneal injection of cyclosporin A, a potent inhibitor of calcineurin, induced extensive inflammation, marked fiber atrophy, the appearance of immature myotubes, and calcification in the regenerating muscle compared with phosphate buffered saline-administered mice. Thus, calcineurin may have an important role in muscle regeneration in association with NFATc1 and GATA-2. PMID- 12557016 TI - Characterization of Bax and Bcl-2 in apoptosis after experimental traumatic brain injury in the rat. AB - This study was undertaken to fulfill the need for additional data on the dynamics of Bax and Bcl-2 expression in conjunction to the cell death that ensues following experimental brain contusion. Adult Sprague-Dawley rats were subjected to a unilateral experimental controlled cortical contusion and killed at 1, 2, 4, 6 and 10 days post injury (dpi). Cell death was examined by the terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) method together with immunohistochemistry for cellular markers. Expression of Bax and Bcl-2 were analyzed by immunohistochemistry and in situ hybridization. The number of TUNEL positive cells was highest at 1 dpi and decreased with time. At all time points, 10-16% of the TUNEL-positive cells showed an apoptotic nuclear morphology. The apoptotic features were restricted to neurons and some inflammatory cells. Immunohistochemistry for Bax revealed a translocation of Bax from a diffuse to a granular distribution in neurons. An up-regulation of Bax mRNA at 6 dpi was discernible. This increase was associated with a statistically significant increase in number of cells with up-regulated and translocated Bax protein. Moreover, a statistically significant increase of Bcl-2 mRNA was detected at 10 dpi. The potential window for anti-apoptotic treatment to salvage neurons is wide. The susceptibility of neurons to necrosis and apoptosis through different pathways during a prolonged post-traumatic period indicate that different pharmacological strategies may be required at different time points after trauma. PMID- 12557017 TI - Eosinophilic myelitis associated with atopic diathesis: a combined neuroimaging and histopathological study. AB - Histologically proven eosinophilic myelitis has rarely been reported except in connection with parasitism. To clarify its clinicopathological features, we conducted a nationwide survey of biopsy-proven eosinophilic myelitis of unknown cause throughout Japan. Six such cases were collected and studied immunologically and pathologically. All were young to middle-aged men. All showed a protracted and fluctuating course with mild disability for 3-25 (mean 12.5) months before biopsy. Magnetic resonance imaging revealed localized lesions of T2-high and T1 iso signal intensity with a partial gadolinium enhancement in all cases. Cerebrospinal fluid (CSF) examinations were completely normal except for modest pleocytosis in two cases. Eosinophilia was present in the peripheral blood in two cases but was absent from the CSF of all cases. In spite of the chronic nature of the disease, spinal cord pathology revealed very active lesions with marked cell infiltration consisting mainly of CD8(+) T cells and varying numbers of eosinophils in the perivascular areas and the parenchyma. Both the myelin and axons were severely disrupted in all cases. Moreover, eosinophil cationic protein (ECP), an activated eosinophil product, was heavily deposited in the tissues. All but one case had hyperIgEemia and mite antigen-specific IgE in the sera, and two had accompanying atopic disorders. The present study thus revealed idiopathic eosinophilic myelitis to be a localized and persistent inflammation of the spinal cord, with distinct clinicopathological features, that has a possible link to atopic diathesis. PMID- 12557018 TI - Immunohistochemical localization of 14.3.3 zeta protein in amyloid plaques in human spongiform encephalopathies. AB - The localization of 14.3.3 proteins was studied in different subtypes of brain amyloid plaques. We examined paraffin-embedded brain sections of sporadic MV2 Creutzfeldt-Jakob disease (sCJD) with Kuru plaques, sporadic VV2 CJD with plaque like PrP(sc) (the abnornal form of prion protein) deposits, variant CJD (vCJD) with florid plaques, Gerstmann-Straussler-Scheinker (GSS) with multicentric plaques and of Alzheimer's disease (AD) with senile plaques. Adjacent immunostaining revealed PrP(sc) and 14.3.3 zeta deposits in the same amyloid plaques in all cases of sporadic CJD and vCJD, whereas 14.3.3 zeta was not seen in amyloid plaques of GSS with A117V, P102L and D202N mutations. The same immunostaining method using anti-betaA4 and anti-14.3.3 zeta antibodies revealed no colocalization in patients with AD. Our data suggest that 14.3.3 zeta protein could interact either with PrP or with other components of PrP(sc) deposits in CJD. PMID- 12557019 TI - Papillary endothelial hyperplasia associated with cortical dysplasia. AB - We report a unique case of papillary endothelial hyperplasia (PEH) presenting as a subcortical mass lesion intimately associated with focal cortical dysplasia (CD) and consider a possible causal relationship. A 6-year old girl presented with a 6-month history of a painless, frontoparietal skull "bump" associated with slowly progressive localised bossing followed by a 4-month history of absence attacks. Magnetic resonance imaging (MRI) revealed an adjacent parietal enhancing mass lesion beneath abnormal appearing cortex. A haemorrhagic vascular lesion with histology consistent with that of papillary endothelial hyperplasia was completely resected. Biopsies of the adjacent cortex showed CD. The patient has been symptom free post-surgery for 12 months with no MRI evidence of recurrence. Intracranial PEH is very rare and, in contrast to extracranial examples, half of the reported cases lacked a demonstrable vascular origin. Given that CD may be associated with intrinsic capillary hypervascularity, vascular malformations and tumours (e.g. dysembryoplastic neuroepithelial tumour) of a potential hypervascular or haemorrhagic nature, the association between PEH and CD may not be incidental. The abnormal vascularity not uncommonly found in CD may predispose to haemorrhage and/or thrombosis, the organisation of which may rarely be complicated by PEH. Alternatively, PEH and CD may both represent local, independent complications of a pre-existing vascular event or trauma. PMID- 12557020 TI - [Diagnosis and follow-up in glaucoma patients using the Heidelberg retina tomograph]. AB - The development of laser scanning tomography in the late 1980s enabled the possibility of an exact three-dimensional biomorphorphometry of the optic nerve head. This technique is designed for 3D measurement of the topography of the optic disc with high accuracy and reproducibility. With the development of the Heidelberg retina tomograph with highly advanced and user-friendly software, a quick examination is possible. Currently the instrument is already used on a routine basis in the ophthalmological practice. It has been shown that glaucomatous changes of the optic disc can be detected using laser scanning tomography before perimetric deterioration occurs. Therefore this technique is crucial in the follow-up of glaucoma patients. PMID- 12557021 TI - [Glaucoma diagnosis of the optic disc using photography and planimetry]. AB - Ophthalmoscopically and on clinical optic disc photographs, the optic nerve head can be described by the following parameters: size and shape of the optic disc, size and shape of the neuroretinal rim, size and shape of the optic cup, size and shape of the alpha and beta zones of parapapillary chorioretinal atrophy, visibility of the retinal nerve fiber layer, diameter of the retinal arteries, presence and location of optic disc hemorrhages. PMID- 12557022 TI - [Glaucoma diagnosis with the GDx and measurement of nerve fibre thickness (RTA)]. AB - Damage of retinal nerve fibres is an early sign of glaucoma.Therefore, a quantitative determination of nerve fibres would be an ideal technique for early glaucoma diagnosis. This can be realized by the GDx which uses nerve fibre polarimetry to measure the thickness of the parapapillary nerve fibre layer.A single GDx measurement in a glaucoma patient may be an important adjunct to the clinical findings.However, at present the method is not suitable for general screening. Presumably the most important application of the device is longitudinal monitoring. In contrast to the GDx,RTA determines the thickness of the whole retina, not specifically the nerve fibre layer. This might be a reason why the discrimination between normal subjects and glaucoma patients is somewhat more difficult than with the GDx.Again, longitudinal monitoring should be the most common application of the RTA in glaucoma patients. Besides glaucoma,RTA also allows examination of macular pathologies.RTA is a promising new technique which so far is not as frequently used as the GDx or the HRT. PMID- 12557023 TI - [Reading ability after photodynamic therapy (PDT) for age-related macular degeneration (AMD) and for high myopia]. AB - BACKGROUND: A prospective uncontrolled follow-up of changes in reading ability after PDT with Verteporfin in patients with predominantly classic subfoveal choroidal neovascularization (CNV) due to AMD or high myopia was carried out. PATIENTS AND METHODS: A follow-up time of at least 6 months is documented for 48 patients with AMD and for 22 patients with high myopia. In addition to the usual clinical parameters the need for magnification was measured using standardized reading charts provided by the SZB.A need for magnification higher than 3.2-fold was defined as a severe loss in reading ability.PDT's were repeated every 3 months according to the usual guidelines. RESULTS: Before treatment, the mean reading acuity for patients with AMD was between 20/200 and 20/100. The average need for magnification was between 2.5-fold and 3.2-fold.Reading acuity, need for magnification and the percentage of patients with a need for magnification of 3.2 fold or less remained stable over a 6-12 months follow-up period. High myopic patients initially had a mean reading acuity of 20/200 and a need for magnification of 2.5-fold. After 9 months follow-up, reading acuity showed a mean improvement from 0.2 up to 0.3 ( p<0.05). There was also a trend towards improvement in the need for magnification and the percentage of patients with a need for magnification of 3.2-fold or less improved significantly from 68% to 78% ( p<0.05). CONCLUSION: Reading ability could be stabilized with PDT for predominantly classic subfoveal CNV in patients with high myopia or AMD for at least 9-12 months. The risk of developing a severe loss in reading ability (need for magnification >3.2-fold) was significantly reduced in high myopia and was stable over time in AMD. PMID- 12557024 TI - [Pars plana vitrectomy in cystoid macular edema of different forms of chronic uveitis]. AB - BACKGROUND: Cystoid macular edema (CME) is a common complication in different forms of chronic uveitis. In spite of immunosuppressive and anti-inflammatory therapy, chronic or relapsing courses can occur which may have a negative impact on visual prognosis. Pars plana vitrectomy (PPV) is known to positively influence chronic uveitis. This retrospective study was performed to investigate the role of PPV in the therapy refractive uveitic CME. PATIENTS AND METHODS: PPV for CME was performed in eyes with CME in intermediate uveitis (IMU, n=42), chronic iridocyclitis in juvenile rheumatoid arthritis (CIC, n=14) and multifocal chorioretinitis (MFC, n=12). In none of the eyes had immunosuppressive and/or anti-inflammatory therapy or anti-edema treatment (e.g.acetazolamide) led to regression of the CME. After a postoperative follow-up period of 7 and 106 months all patients were re-examined. RESULTS: Postoperative complete or partial regression of CME was observed as follows: IMU: 25/42 (59.5%), CIC: 8/14 (57.1%),MFC: 5/12 (41.7%). A significant increase in visual acuity of 2 lines and more was observed in 50%,71.4% and 41.7% of eyes, respectively. In the long-term follow-up best functional results were achieved in eyes with IMU. CONCLUSIONS: Response to PPV was variable according to the type of underlying form of uveitis. The lowest success rate could be observed in eyes with MFC. Although the postoperative regression rate of CME was satisfactory in eyes with CIC, long-term visual acuity results were disappointing due to secondary complications of CIC in this young age group. Best results were achieved in patients with IMU (statistically not significant). A multicenter study in a larger series of patients is needed to investigate the exact role of PPV in different forms of chronic uveitis. PMID- 12557025 TI - [Treatment of keratoconus by collagen cross linking]. AB - BACKGROUND: We were able to show a significant increase in corneal stiffness of rabbit and porcine eyes after combined riboflavin/UVA-induced collagen cross linking. In this study,we tried to treat keratoconus patients with this method to stop the progression of corneal ectasia. PATIENTS AND METHODS: We treated 16 eyes of 15 patients with progressive keratoconus and mostly moderate keratectasia (48 56 dpt). After removal of the epithelium (7 mm X), riboflavin solution was applied on the cornea, which was irradiated with UVA (370 nm,3 mW/cm(2)) at a distance of 1 cm for 30 min.Post-operative follow-up controls were conducted every 3 months in the first year and then every 6 months, always including visual acuity testing, corneal topography and measurements of endothelial cell density. The follow-up time was between 1 and 3 years. RESULTS: Progression of keratectasia was stopped in all patients. Best corrected visual acuity and the maximal keratometry values improved slightly in about 50% of the cases. In all patients corneal transparency, the degree of keratectasia registered by corneal topography and the density of endothelial cells remained unchanged within the follow-up time. No negative side-effects were observed. CONCLUSIONS: Our results show that collagen cross linking might be a useful conservative treatment modality to stop the progression of keratoconus. By this means the need for keratoplasty might be significantly reduced. Given the simplicity of the technique and minimal costs of the treatment it might also be well suited for developing countries.Further studies are envisaged to exclude long-term side effects and to evaluate the long term durability of the mechanical stiffness effect. PMID- 12557027 TI - [Hyperornithinaemia in patients with retinal dystrophy]. AB - BACKGROUND: In our clinic for low vision we have treated 4 patients with advanced retino-choroidal dystrophy. The patients showed concentric narrowing of the visual field and alterations of the ERG.Confluent degenerative areas slightly resembled those seen in gyrate atrophy.For this reason we determined the serum levels of ornithine which were found to be elevated to twice the normal level, but never as high as described for the typical form of gyrate atrophy. CASE HISTORIES: The patients were three females 58-76 years old and the 11-year-old grandson of one of them. We analysed ornithine levels in the corresponding blood serum. RESULTS: The results were 2.8 mg/dl, 2.0 mg/dl, 2.6 mg/dl and 3.6 mg/dl, respectively (normal value: 1.8 microg/dl). CONCLUSION: In classic gyrate atrophy the ornithine levels are much higher. Consequently we are dealing with a biochemically and symptomatically different disease or with a mild form of gyrate atrophy. Narrowing of the visual field and ERG alterations indicate a retinal dystrophy. The determination of ornithine levels does not yet belong to our clinical routine. This report is intended to enlarge the possibilities for differential diagnosis and to offer a therapy for the 11-year-old boy by giving him vitamin B6 or a special diet. PMID- 12557028 TI - [Nobel Prize 2002. Organ development and programmed cell death]. PMID- 12557026 TI - [Development of dynamic stereopsis after eye muscle operations for binocular vision impairment]. AB - BACKGROUND: Strabism usually impairs central stereopsis irreversibly after a short time duration,however, the retinal periphery is more resistant to deterioration by binocular impairment. METHODS: In this prospective study 46 strabismic patients (9-56 years old) were tested for dynamic stereopsis in the peripheral visual field up to 20 degrees eccentricity. The effect of realignment of the visual axes by surgery on dynamic stereopsis was tested before and after surgery in 40 out of these patients. Patients were tested qualitatively for dynamically stereoactive fields of vision and quantitatively for the threshold value needed to create a three-dimensional impression. RESULT: We found residual dynamic stereopsis in 30% of patients without central static stereopsis: 56% of the patients improved after surgery, either through a significant (p<0.01) gain of stereoactive fields or through a decrease of threshold values. CONCLUSION: Strabismus surgery has benefits for the patients outside classical tests and seems to be independent of the static depth perception. PMID- 12557029 TI - [Ambliopia. Part 1:diagnosis]. PMID- 12557030 TI - Intrapulmonary expression of scavenger receptor and CD14 and their relation to local inflammatory responses to endotoxemia in mice. AB - This study was first designed to investigate systematically the kinetics of surface expression of scavenger receptors (SRs) and CD14 on alveolar macrophages in vivo and in vitro and their relation with local pro- and antiinflammatory responses in endotoxemia. The expression of SR and CD14 in lungs was down- and up regulated, respectively, in the presence of endotoxemia, which might be due to decreased expression of SR and increased expression of CD14 on the surface of the resident macrophages. Down-regulation of SRs on alveolar macrophages not only induces decreased defensive function of the macrophages, it also enhances lipopolysaccharide (LPS)-induced activation of alveolar macrophages possibly through increasing LPS binding to CD14. Although CD14 is a key receptor responsible for LPS to activate macrophages, both phospholipase C and anti-CD14 antibody can completely inhibit activation of alveolar macrophages initiated by only LPS 1 ng/ml, as determined by tumor necrosis factor-alpha (TNFalpha) production, but it does not significantly change TNFalpha release upon cell stimulation by LPS 10 microg/ml. There was an intrinsic relation of enhanced intrapulmonary pro- and antiinflammatory responses with changes in SR and CD14 expression, which suggests that the down-regulation of SR and up-regulation of CD14 might be an important mechanism for the lung to change from a defense organ to an effector organ during sepsis. PMID- 12557031 TI - Circulating soluble Fas in patients with breast cancer. AB - It has been suggested that circulating soluble Fas (sFas) contributes to tumor progression. However, little is known about the role of sFas in breast cancer. This study was designed with the aim of elucidating the possible relation between sFas and breast cancer. A series of 57 consecutive patients with invasive breast cancer undergoing surgery were prospectively included in the study and evaluated. Venous blood samples were collected before surgery. Sera were obtained by centrifugation and stored at -70 degrees C until assayed. The control group consisted of 12 patients with benign breast tumors (6 with fibrocystic disease, 6 with fibroadenoma). Serum concentrations of sFas were measured by the quantitative sandwich enzyme immunoassay technique. The data on primary tumor staging, age, estrogen receptor status, lymph node status, tumor grading, and TNM staging were reviewed and recorded. The mean value of circulating sFas in patients with invasive breast cancer was 794.2 +/- 183.0 pg/ml and that of the control group 582.1 +/- 62.8 pg/ml; the difference was significant (p < 0.001). Furthermore, there were significantly higher serum levels of sFas in the older patients (age > or = 50) (p = 0.020) and in those with a more advanced TNM stage (p = 0.021). In the multivariate analysis, TNM stage (p = 0.005) appeared to be an independent factor for significantly higher circulating sFas in patients with invasive breast cancer. Thus circulating sFas levels may reflect the severity of invasive breast cancer. Hence the possible prognostic value of sFas for breast cancer deserves further elucidation and evaluation with long-term patient follow up. PMID- 12557032 TI - Extracapsular hematoma of the parathyroid glands. AB - The objective of this study was to standardize surgical treatment of cervicothoracic hematoma due to parathyroid gland rupture. Only 19 such hematomas have been reported in the literature, and there is no consensus about the best time to operate or the surgical approach. We have now treated four new cases of extensive hematoma from the time of the initial bleeding except in the case of severe hemodynamic or respiratory troubles or nerve compression. Two of the patients were operated on after a minimal 3 months delay, with perfect results. The other two were operated on during the first month with great technical difficulty and incomplete results. In one case the gland was not excised, but parathyroid apoplexy afforded a spontaneous remission, although the hyperparathyroidism recurred 7 years later. We concluded that, first, if there are no severe compressive or hemodynamic symptoms, the surgical treatment must be performed more than 3 months after the bleeding, as the dissection then is as simple as any well ordered surgery. In case of an extensive hematoma accompanied by shock or compression, we propose simple emergency drainage, with reoperation 3 months later. Second, parathyroid apoplexy sometimes offers spontaneous remission of primary hyperparathyroidism, although late recurrence is always possible and surgical treatment then cannot be avoided. PMID- 12557033 TI - Paul of Aegina: landmark in surgical progress. AB - During the Byzantine period the most prominent medical personalities were Oribasius, Aetius of Amida, Alexander of Tralles, and Paul of Aegina (Paulus Aegineta). The last of the eclectic Greek compilers, Paul of Aegina (625-690 AD) was born on the island of Aegina and practiced medicine in Alexandria. He was the author of the Epitome of Medicine (seven books), which was first printed in Greek by the Aldine Press in Venice in 1528. The Syndenham Society of London published an English translation by Francis Adams of Banchory between 1844 and 1847. The most noteworthy of his Epitome is the sixth book on surgery. Paul was not only a scribe but also a highly capable surgeon. He was the quintessential student of the best medical authorities: Hippocrates and Galen in Greek and Roman medicine, respectively. He also displayed a peculiar genius in the field of surgery. He gave us novel descriptions of tracheotomy, tonsillectomy, catheterization of the bladder, lithotomy, inguinal hernia repair, abdominal paracentesis for ascites, and many other surgical procedures including reduction of breast size. He not only influenced those in his own era but had great influence on physicians such as Rhazes, Haly Abbas, Albucasis, Avicenna, and Fabricius ab Aquapendente, who lived in subsequent eras. This historical article emphasizes the role of Paul of Aegina in the history of surgery and provides a comprehensive review of his surgical treatise with original case examples that represent his contributions to surgical progress. PMID- 12557035 TI - Prenatal diagnosis. PMID- 12557036 TI - Organogenesis particularly relevant to fetal surgery. AB - In utero surgical intervention is an exciting frontier in medicine. Fetal surgeons strive to treat congenital anomalies definitively while organogenesis is still occurring. Many of these anomalies pose such a threat to the viability of the affected fetus that waiting until after the child is born to treat them is frequently not satisfying and too often unsuccessful. We review the embryology of selected systems that have associated aberrancies of development for which fetal surgery is particularly applicable. The surgeon can more effectively launch an assault against congenital anomalies when armed with a solid appreciation of normal development. Recognizing the critical period for the development of a system allows him or her to formulate the optimal time and mode of intervention. PMID- 12557037 TI - Pathophysiologic patterns influencing fetal surgery. AB - There are a growing number and variety of fetal disorders that may benefit from intervention prior to birth. Despite the diversity, there are common pathophysiologic denominators or patterns that tie together many seemingly disparate disorders. The purpose of this article was not to review disorders presented in other, accompanying articles but, rather, to present pathophysiologic patterns that common influence fetal surgery within a cohesive framework. PMID- 12557038 TI - Fetal wound healing: current biology. AB - The early-gestation fetus heals dermal wounds rapidly and scarlessly. This phenomenon appears to be intrinsic to fetal skin and independent of the intrauterine environment. Unique properties of fetal cells, extracellular matrix, cytokine profile, and gene expression contribute to scarless repair. An intensive research effort has focused on unraveling the mechanisms that underlie scarless fetal wound healing in an attempt to improve the quality of healing in both children and adults. PMID- 12557039 TI - Prenatal therapy for obstructive uropathy. AB - The increasing use of routine prenatal imaging and diagnosis has provided a better understanding of the natural history of obstructive uropathy. Fortunately, most disorders portend little clinical significance prenatally, although a select subset can incur irreversible renal injury and pulmonary hypoplasia depending on the onset, duration, severity, and level of obstruction. In these cases, timely antenatal evaluation and prenatal intervention can provide an optimal postnatal outcome. The natural history, evaluation, and management of fetal obstructive uropathies are discussed. PMID- 12557040 TI - Congenital diaphragmatic hernia: advances in prenatal therapy. AB - Congenital diaphragmatic hernia (CDH) is one of the most common causes of neonatal morbidity and mortality. The clinical spectrum of CDH ranges from minimally affected infants who do well with modern neonatal care to severely affected infants who die despite all interventions. Two decades of research have led to advances in the prenatal diagnosis of CDH and have better defined the natural history of CDH. Fetuses with CDH now can be stratified into "low" and "high" risk groups based on sonographic parameters. "Low risk" fetuses have an excellent chance of survival with postnatal therapy. Prenatal intervention is reserved for "high risk" fetuses. Ongoing research is focused on improving both prenatal and postnatal treatment of these severely affected infants. PMID- 12557041 TI - Prenatal therapy for thoracic and mediastinal lesions. AB - Most prenatally diagnosed lung lesions can be managed successfully during the neonatal period. Prenatal imaging and experimental models have provided a comprehensive understanding of the pathophysiology, natural history, and prognosis of intrathoracic and mediastinal lesions. Clinical experience has demonstrated that progression to nonimmune hydrops fetalis and pulmonary hypoplasia is a harbinger of fetal or neonatal demise. Advances in fetal anesthesia, tocolysis, and surgical techniques have made fetal surgery a viable in utero option to ameliorate life-threatening masses. Congenital cystic adenomatoid malformation, bronchopulmonary sequestration, and congenital hydrothorax are the most common abnormalities amenable to surgical intervention. The natural history, evaluation, and treatment of intrathoracic and mediastinal lesions are discussed. PMID- 12557042 TI - Fetal sacrococcygeal teratoma. AB - Advances in prenatal diagnosis including improvements in ultrasonography have revealed the natural history of fetal sacrococcygeal teratomas, and this natural history differs substantially from that for postnatally diagnosed sacrococcygeal teratoma. A fetal sacrococcygeal teratoma may lead to perinatal morbidity and mortality by a variety of mechanisms. Adverse clinical sequelae of a sacrococcygeal teratoma can be prevented by accurate prenatal assessment and appropriate obstetrical and perinatal management. Development of fetal hydrops and/or placentomegaly predicts fetal demise. Fetal surgical intervention has proven successful in highly selected cases. PMID- 12557043 TI - Fetal surgery for myelomeningocele: panacea or peril? AB - Myelomeningocele affects thousands of children worldwide with devastating consequences. In an effort to improve neurologic outcome, fetal surgery has been performed for myelomeningocele for the past 5 years. Sensorimotor function is not appreciably improved, although there may be a reduction in hindbrain herniation and a decreased need for ventriculoperitoneal shunting. The long-term clinical consequences of these findings are not clear. What is clear, however, is that further study in the form of a prospective, randomized trial is mandatory. PMID- 12557044 TI - Minimal access techniques for fetal surgery. AB - The impetus for the development of minimal access techniques for fetal surgery was the unusual occurrence with open hysterotomy of preterm labor, premature rupture of membranes, and the maternal complications resulting from tocolytic therapy. This strategy involves a constellation of techniques that allow surgical procedures to be performed inside the uterus without a hysterotomy. The unique requirements of this approach necessitated modifications of existing endoscopic techniques, development of novel fetoscopic instruments, and utilization of the expertise of a wide variety of specialists. Technical expertise in the field and a natural evolution of techniques have given rise to innovative repairs previously not envisioned. Severe congenital diaphragmatic hernia, diseases of monochorionic twins, and obstructive uropathy have already been successfully treated using fetoscopic surgical techniques. Fetoscopic correction of many other non-life-threatening anomalies continues to evolve. The future of fetoscopic surgical intervention depends on the continual evolution of novel techniques, the elucidation of the pathophysiology and treatment of other fetal disorders, and a better understanding of treatment of complications of intervention. PMID- 12557045 TI - Fetal intervention for complications of monochorionic twinning. AB - All multiple gestation pregnancies have a significantly higher risk of fetal morbidity and mortality than singleton pregnancies. The risks become progressively greater as the number of fetuses increases. Among twin pregnancies, there is a greater danger of an adverse outcome when the twins share a single placenta (monochorionic), than if each has a separate placenta (dichorionic). A still greater danger exists when monochorionic twins also share a single amniotic sac. Fetal surgical interventions now exist to treat many of the complications associated with twin gestation pregnancies. PMID- 12557046 TI - In utero models of craniofacial surgery. AB - Fetal surgery is finding ever-increasing applications. At present, fetal surgical techniques are used only to treat diseases associated with high mortality risk or severe morbidity if left untreated prior to birth. Yet as a discipline, in utero therapy is attractive to plastic surgeons because it affords the potential to (1) provide a scarless repair, (2) correct the primary deformity, (3) prevent secondary deformities, and (4) give the parents a "normal"-appearing child at birth. Each of these potential benefits is particularly important in patients with craniofacial anomalies. This article will both review the animal models that have been used to study the potential for intrauterine plastic surgery and provide a synopsis of the benefits that fetal therapy may provide in the treatment of selected craniofacial anomalies. PMID- 12557047 TI - Abdominal wall defects. AB - The frequent use of prenatal diagnostic techniques including ultrasound and maternal serum alpha-fetoprotein has increasingly led to detection of abdominal wall defects before birth. This prenatal detection creates the opportunity to influence neonatal outcome by alteration in management of pregnancy or delivery. The optimal management of an individual fetus depends on careful prenatal assessment of the abdominal wall defect, combined with experience and knowledge of the natural history for that particular lesion. A multidisciplinary approach to the fetus can improve neonatal outcome. Careful assessment for other structural anomalies and karyotype analysis should be performed. Delivery at a high-risk perinatal center should be encouraged. Currently, there is no convincing evidence to support routine cesarean section for most abdominal wall defects. PMID- 12557048 TI - Extratemporal hypometabolism on FDG PET in temporal lobe epilepsy as a predictor of seizure outcome after temporal lobectomy. AB - We investigated the relationship between the presence of extratemporal hypometabolism on fluorine-18 fluorodeoxyglucose positron emission tomography (FDG PET) and seizure outcome after temporal lobectomy in patients with medically intractable temporal lobe epilepsy (TLE). In 47 patients with intractable unilateral mesial TLE, regional metabolic changes on FDG PET images obtained during the 2 months preceding anterior temporal lobectomy were compared with postoperative seizure outcome. Postoperative seizure outcome was evaluated with a mean follow-up period of 6.1+/-0.6 years (range 5.2-7.2 years). Forty-two (89%) of the 47 patients achieved a good postoperative seizure outcome (Engel class I or II). All patients had hypometabolism in the temporal cortex ipsilateral to the epileptogenic region on FDG PET scans. Fourteen (78%) of the 18 patients with hypometabolism only in the ipsilateral temporal cortex were completely seizure free (Engel class Ia) after surgery. In contrast, five (45%) of the 11 patients with extratemporal cortical hypometabolism confined to the ipsilateral cerebral hemisphere and only four (22%) of the 18 patients with hypometabolism in the contralateral cerebral cortex were completely seizure free after surgery. The postoperative seizure-free rates were significantly different across the three groups of patients with different cortical metabolic patterns ( P<0.005). Furthermore, all of the nine patients with a non-class I outcome (Engel class II IV) had extratemporal (including contralateral temporal) cortical hypometabolism. Thalamic hypometabolism was noted in 20 (43%) of the 47 patients (ipsilateral in 12, bilateral in 8). Sixteen (59%) of the 27 patients with normal thalamic metabolism were completely seizure free after surgery, while only seven (35%) of the 20 patients with thalamic hypometabolism became completely seizure free ( P<0.05). Multivariate analysis revealed that among variables including clinical, EEG, magnetic resonance imaging, pathological and FDG PET metabolic findings, only cortical metabolic pattern was an independent factor for the prediction of postoperative seizure outcome ( P<0.005). It is concluded that extratemporal cortical hypometabolism outside the seizure focus, in particular hypometabolism in the contralateral cerebral cortex, may be associated with a poorer postoperative seizure outcome in TLE and may represent underlying pathology that is potentially epileptogenic. Thalamic hypometabolism, which was associated, but not independently, with a higher likelihood of postoperative seizures, may be secondary to extratemporal or temporal pathology. PMID- 12557049 TI - Radioimmunotherapy: is avidin-biotin pretargeting the preferred choice among pretargeting methods? PMID- 12557050 TI - Do we need more nuclear cardiologists in Europe? PMID- 12557051 TI - Do we need more nuclear cardiologists in Europe? PMID- 12557052 TI - Specific endothelial binding and tumor uptake of radiolabeled angiostatin. AB - Angiostatin (AS) is a potent antiangiogenic agent which inhibits tumor growth through specific action on proliferating endothelial cells. Imaging of radiolabeled AS would enhance our knowledge on the pharmacokinetics of AS and might provide useful information relating to tumor neovasculature. We therefore investigated the potential of radiolabeled AS as a novel tumor imaging agent. Human angiostatin was radioiodine labeled using the lactoperoxidase method. Competition binding studies showed a dose-dependent inhibition of (125)I-AS binding to endothelial cells by excess unlabeled AS, and a displacement curve demonstrated that specific binding was dose dependent and saturable, with a K(d) value of 169 n M. Gel analysis showed that (125)I-AS remained stable in serum for up to 24 h without significant degradation. Intravenously injected (125)I-AS in rats was cleared from the blood in an exponential fashion. Biodistribution data from human colon cancer-bearing Balb/C nude mice showed high uptake in the kidneys, stomach, liver, and lungs. Tumor uptake was 3.2+/-0.7, 2.6+/-0.2, and 1.7+/-0.2%ID/g at 2, 4, and 9 h after injection, respectively. Tumor to muscle count ratio increased from 3.1+/-0.5 at 2 h to 4.4+/-0.5 at 9 h. Serial scintigraphy from 1 to 5 h after (123)I-AS injection demonstrated high uptake in the kidneys and bladder, consistent with renal excretion. There was clear demarcation of tumor by 1 h, with gradual increase in contrast over time (4-h tumor to contralateral thigh ratio =4.7+/-1.1). Thus, radioiodine-labeled angiostatin binds specifically to endothelial cells and has potential as a novel tumor imaging agent. PMID- 12557053 TI - Functional analysis of a promoter variant of the gene encoding the interferon gamma receptor chain I. AB - We analyzed a single nucleotide polymorphism (SNP) at position -56 (T-->C) in the promoter region of the gene encoding the human interferon-gamma receptor ligand binding chain I (IFN-gammaR1). The mutation was present at similar frequencies in Gabonese children with either mild or severe malaria. Functional investigations of the promoter in a transfected human B-cell line showed lower levels of luciferase reporter gene expression in the presence of the mutation, indicating the importance of this position for promoter activity, and suggesting that this SNP might negatively influence the expression level of IFN-gammaR1 at the cell surface. Further examinations of the DNA sequence at this polymorphic site showed a perfectly matched binding site for the transcription factor activator protein 4 (AP-4) on both strands. Binding sites for other important transcription factors involved in gene expression and regulation of the immune response against infections, including Ikaros 2 (Ik-2), nuclear factor kappaB (NFkappaB), and CETS1p54, are also situated in this region. PMID- 12557054 TI - Two genes, three messengers: hybrid transcript between a gene expressed at specific stages of T-cell and sperm maturation and an unrelated adjacent gene. AB - To identify new genes that are turned on following the expression of a pre-T-cell antigen receptor in early thymocytes, we developed a PCR-based subtractive approach that selects for mRNAs that are differentially expressed between thymocytes at the CD4(-)CD8(-) and CD4(+)CD8(+) stages. This approach led to the characterization of a cDNA, Tssn1, which is predominantly expressed in CD4(+)CD8(+) cells and at specific stages of spermatogenesis. We analyzed the structure of the Tssn1 gene in mouse and human and found that it is highly conserved. Of interest, Tssn1 is adjacent to an unrelated gene named 2310047L21Rik and gives rise to hybrid 2310047L21Rik--Tssn1transcripts. Therefore, this analysis offers insights into the critical steps that may precede the birth of new genes endowed with either compound functions or novel tissue distribution. PMID- 12557055 TI - The donor splice site mutation in NFkappaB-inducing kinase of alymphoplasia (aly/aly) mice. AB - The alymphoplasia (aly/aly) mouse has a spontaneous mutation maintained on a C57BL/6xAEJ ( H-2(b)) background that results in an absence of extrasplenic secondary lymphoid tissues. The cDNA defect has previously been shown to reside in a point mutation causing a G855R substitution in NFkappaB-inducing kinase (NIK). Since the aly/aly female cannot lactate, the strain must be bred by intercrossing heterozygous females with homozygous males and the offspring typed by serum IgA levels at the age of 4-6 weeks. We originally determined the genomic location of the alymphoplasia mutation by sequencing boundaries of regions homologous to human NIK exons, although recently the entire genomic sequence of murine C57BL/6 NIK has become available through the mouse genome project. The aly mutation is at position -1 of an intron donor consensus splice site. Exon connexion PCR confirmed that splicing does occur across this site. Using the genomic information, we also developed a method of PCR typing of aly/aly mice from tail clips, and used this to derive an aly/aly muMT double-mutant strain in which antibody independent typing is essential. Genetic typing should considerably simplify husbandry and manipulation of the aly/aly genetic background, which is widely used as a recipient in lymphocyte transfer experiments to permit examination of the relative role of secondary lymphoid structures in immune responses. PMID- 12557056 TI - Comparative and evolutionary analysis of the rhesus macaque extended MHC class II region. AB - The sequence-based map of a part of the rhesus macaque major histocompatibility complex (MHC) extended class II region is presented. The sequenced region encompasses 67,401 bp and contains the SACM2L, RING1, FABGL and KE4 genes, as well as the HTATSF1-like and ZNF-like pseudogenes. Similar to human, but different from rat and mouse, no class I genes are found in the SACM2L- RING1 interval. The rhesus macaque extended MHC class II region shows a high degree of conservation of exonic as well as intronic and intergenic sequences compared with the respective human region. It is concluded that this particular genomic organization of the extended class II region-i.e., the absence of class I genes and the presence of the HTATSF1-like and ZNF-like pseudogenes-can be traced back to a common ancestor of humans and rhesus macaques about 23 million years ago. PMID- 12557057 TI - Molecular cloning of the pufferfish (Takifugu rubripes) Mx gene and functional characterization of its promoter. AB - Mx proteins are members of a family of interferon-inducible genes that are expressed by cells in response to viral infection. They are important determinants of innate immunity against viral infection in vertebrates. We cloned the pufferfish ( Takifugu rubripes) Mx gene and sequenced 80 kb from the Mx locus. The Fugu Mx gene spans 3.4 kb from the transcription start site to the polyadenylation signal, and is made up of 12 exons and 11 introns. The protein sequence encoded by the Fugu Mx gene is 77%, 48%, and 51% identical to that of trout Mx1, chicken Mx, and mouse Mx1 genes, respectively. The Fugu Mx gene is expressed in a variety of tissues, with high expression detected in the heart, gill, kidney, intestine, and brain. Analysis of the 5'-flanking sequence of the gene showed the presence of two interferon-stimulated response elements (ISRE) at positions -51 to 38 and -97 to 85, relative to the transcription start site. The Fugu Mx promoter was inducible by human IFN-beta in the human hepatoma (Huh7) cells and by polyinosinic: polycytidilic acid in the top minnow hepatoma (PLHC-1) cells. Deletion analysis of the promoter showed that both ISREs contributed to inducibility. These results demonstrate that the molecular mechanisms involved in Mx gene regulation are conserved between fish and mammals. PMID- 12557058 TI - Molecular characterization of the pig C3 gene and its association with complement activity. AB - The complement system catalyzes direct lysis of micro-organisms and modulates phagocytosis, inflammation, humoral and cellular immune responses. Since the complement protein C3 is the central component within all pathways of complement activation, C3 is a candidate gene for complement activity and also for improved protection against many pathogens. The pig C3 gene was sequenced, screened for polymorphisms, and analyzed for association with hemolytic complement activity of the alternative and classical pathway (AH(50), CH(50)). C3c serum levels and haptoglobin (HP) serum concentrations were measured before and after vaccination against Mycoplasma hyopneumoniae, Aujeszky virus, and porcine reproductive and respiratory syndrome virus in F2 animals of a pig resource population based on crossbreeding of Duroc and Berlin Miniature Pig. The genomic C3 sequence covers 444 bp of promoter region, 41 exons and 40 introns, as well as 881 bp of the 3' flanking region. The cDNA codes for a 1,661-amino acid precursor C3. Five polymorphic sites were detected in the 5'-UTR, intron 13, exon 15, exon 30, and the 3'-UTR. Within the resource population two haplotypes were found to segregate. Analysis of variance applying a repeated measures model revealed a significant effect of the interaction of C3 genotype and time of measurement relative to immunization on CH(50), AH(50,)and C3c that is likely to be due to variation of C3 expression. In contrast, the time course of the HP acute-phase reaction is not associated with C3 genomic variation. The association of C3 with complement activity indicates the importance of C3 as a candidate gene for natural resistance to micro-organisms, although the causative polymorphism modulating the expression of C3 remains to be delineated. PMID- 12557059 TI - Characterization of the beta2-microglobulin gene of the horse. AB - A clone containing beta(2)-microglobulin (beta(2)-m), the light chain of the major histocompatibility complex class I cell surface molecule, was isolated from an equine bacterial artificial chromosome library. This clone was used as a template for polymerase chain reaction (PCR) and unidirectional sequencing to elucidate the genomic sequence and intron/exon boundaries. We obtained 7,000 bases of sequence, extending from 1,100 nucleotides (nt) upstream of the coding region start through 1,698 nt downstream of the stop codon. The sequence contained regulatory elements in the region upstream of the coding sequence similar to those of the beta(2)-m gene of other species. The beta(2)-m gene was localized to horse chromosome ECA1q23-q25 by fluorescent in situ hybridization. This was confirmed by synteny mapping on a (horse x mouse) somatic cell hybrid panel. The sequence and intron/exon boundaries determined were used to design PCR primers to amplify and sequence the coding region of the beta(2)-m gene in other equids, including five breeds of domestic horse, one Przewalski's horse, five domestic donkeys and five zebras. A high degree of conservation was found among equids, illustrated by >98% (349/354) identity at the nucleotide level and 95% (113/118) at the amino acid level, because of non-synonymous nucleotide substitutions. The promoter detected in the region upstream of the coding sequence was subcloned and used in chloramphenicol acetyl transferase (CAT) assays to demonstrate the presence of a functional promoter. This study provides tools for the analysis of regulation of not only the horse beta(2)-m gene, but also for any genes dependent upon beta(2)-m for expression. PMID- 12557060 TI - Patterns of polymorphism in the MHC class II of a non-passerine bird, the great snipe (Gallinago media). AB - The genomic organisation of the major histocompatibility complex (MHC) seems to vary considerably between different bird species. In order to understand this variation it is important to gather information from different species. We have, for the first time, investigated MHC class II polymorphism in a wader species, the great snipe (Gallinago media). Eleven alleles were found in five sequenced individuals; these come from at least three different loci, but RFLP data suggest that a larger number of genes may be present. For MHC genes, amino acid substitutions followed the, for MHC genes, general pattern of high non-synonymous substitution rates in peptide-binding regions, suggesting that the sequenced alleles may be expressed. The number of genes, lengths of introns and exon sequences of the great snipe MHC seem to be intermediate between those of chicken and passerine birds. PMID- 12557061 TI - Variations in the human Th2-specific chemokine TARC gene. AB - Th2-specific chemokine thymus and activation-regulated chemokine (TARC)/CC chemokine ligand (CCL)17 is highly implicated in the pathogenesis of Th-2 dominated allergic diseases such as bronchial asthma (BA) and atopic dermatitis (AD). We performed polymorphism screening of the coding and promoter regions of the TARC gene. We found two rare variations in the coding region of exon 3 (2134C>T and 2037G>A) and a single nucleotide polymorphism (SNP) in the 5' flanking region (-431C>T). Individuals carrying the 431T allele showed significantly increased serum levels of TARC compared with those not carrying the 431T allele, suggesting that this SNP has functional significance. However, when the genotypes at the SNP site were determined for 158 healthy individuals, 105 patients with BA and 148 patients with AD, we observed no significant association of the SNP with susceptibility to BA or AD. PMID- 12557062 TI - Intussusception. Part 1: a review of diagnostic approaches. AB - The vast majority of symptomatic intussusceptions in children arise in the ileum and are either ileocolic or ileoileocolic. The clinical diagnosis of these "idiopathic" intussusceptions may be difficult to make. Failure to make a prompt diagnosis and initiate appropriate treatment may lead to bowel ischemia, perforation, peritonitis, shock and even death. The clinician, therefore, may have to rely on imaging procedures to diagnose or exclude the presence of intussusception promptly and accurately. The imaging diagnosis of intussusception can be made with sonography or plain abdominal radiographs or by contrast (including air) enema examinations of the colon. This article highlights the current concepts and some controversial issues related to the imaging diagnosis of intussusception. PMID- 12557063 TI - Intraperitoneal fluid in children: normal ultrasound findings depend on which scan head you use. AB - BACKGROUND: The detection of small quantities of unsuspected free intraperitoneal fluid (IPF) at US examination in children has increased markedly in our department since linear high-frequency probes have been used routinely for gastrointestinal examinations. Its clinical significance is unknown. OBJECTIVE. This study attempted to evaluate the sonographic detection, quantification and location of IPF in an unselected paediatric population using both convex or sector scan heads and a linear US probe in all to compare the results obtained using different probes. MATERIALS AND METHODS: We prospectively scanned 272 unselected consecutive patients using a 7- or 5-MHz multifrequency sector or convex scan head and a 7-MHz linear US probe. IPF was searched for and recorded as 0=none, 1=minimal, 2=small, 3=moderate and 4=large amounts in the cul-de-sac, Morison's pouch and between bowel loops. The patient's position on arrival, indication for the sonogram, clinical expectation of IPF and result of the US examination were recorded until 100 children (46 girls, 54 boys; age 3 days to 15 years) with IPF had been seen. Statistical analysis was done comparing all variables one by one, two by two and three by three. RESULTS: Of the 100 children with IPF, its presence was expected in 7 and clinically explainable in 5 others. Nine of these had moderate-to-large amounts of IPF, seen well with all US scan heads. In the other 88 asymptomatic subjects, the detection of unsuspected IPF varied significantly with different probes. Fluid in the cul-de-sac was detected with similar frequency with all probes, whereas fluid between bowel loops was seen much more frequently with the linear probe ( P>0.0000). Position on arrival, age and sex had no influence. CONCLUSIONS: Minimal and small amounts of IPF can be seen between bowel loops in up to 22% of normal children. In this location, it is often seen using a linear scan head only. In the cul-de-sac, a sector scan probe is equally as able to detect IPF as a linear probe. When scanning the bowel with a linear high-frequency US probe, small pockets of mobile anechoic IPF should be considered a normal finding. PMID- 12557064 TI - Contrast-enhanced MR angiography of pulmonary venous abnormalities in children. AB - BACKGROUND: Echocardiography and X-ray angiography have been considered as gold standards for evaluation of pulmonary venous abnormalities. However, each technique has its own limitations, such as limitation in visualization of the pulmonary veins within the lungs by echocardiography, and the invasive nature of and use of ionizing radiation in X-ray angiography. Contrast-enhanced MR angiography (MRA) is a fast noninvasive method of visualization of the vessels including the pulmonary arteries and veins. OBJECTIVES: To evaluate the utility of contrast-enhanced MRA in the evaluation of pulmonary venous abnormalities in pediatric patients and to compare its diagnostic accuracy with that of transthoracic echocardiography. MATERIALS AND METHODS: In 30 pediatric patients 31 contrast-enhanced MRA studies were performed for evaluation of pulmonary venous abnormalities. Each of 124 pulmonary veins was evaluated for site of connection, course within the lung, presence of obstruction, and topographic relationship with the adjacent structures. The findings of MRA were compared with echocardiographic findings for 116 veins in 29 studies in 28 patients. RESULTS: Contrast-enhanced MRA visualized 99% (123 of 124) of the pulmonary veins investigated, while echocardiography visualized 89% (103 of 116). Exact agreement was found between the two methods in 72% of the veins with a weighted kappa of 0.60 (0.47-0.73, 95% CI). Echocardiography failed to diagnose an abnormal connection in 2 of 15 pulmonary veins, a discrete stenosis in 2 of 19 veins, and diffuse hypoplasia in 10 of 14 veins. In 29% of patients, MRA made the uncertain echocardiographic findings clear. In another 29%, MRA provided a new diagnosis. CONCLUSIONS: Contrast-enhanced MRA is a powerful, safe, and accurate fast-imaging technique for the anatomical evaluation of pulmonary venous abnormalities. MRA may obviate the need for conventional X-ray angiography. Cardiac catheterization may be reserved for those patients in whom pulmonary vascular resistance needs to be determined. PMID- 12557066 TI - Three-dimensional volume measurement of the lateral ventricles in preterm and term infants: evaluation of a standardised computer-assisted method in vivo. AB - BACKGROUND: Assessment of ventricular size still represents a challenge for two dimensional cranial sonography. OBJECTIVE: Three-dimensional (3D) volume acquisition and volume quantification are easily accomplished by an automatic scanning system without additional examination time. The development of a computer-assisted ultrasonic device will offer new perspectives for the quantification of ventricular volume. MATERIALS AND METHODS: In total, 250 healthy neonates between 27 and 41 weeks' gestation and infants up to 6 months of age without neurological pathology were examined with 3D cranial ultrasound (US). The volume of both lateral ventricles and the third ventricle were separately quantified and summated for the calculation of ventricular volume. The correlation between body weight (BW), head circumference (HC), gestational age (GA) and ventricular volume was statistically analysed. RESULTS: There was a statistically significant correlation between GA, BW, HC and ventricular volume. Normal values for premature and mature infants were established. CONCLUSIONS: 3D US appears to be an accurate imaging modality for the exact calculation of ventricular volume and should, therefore, be incorporated into the cranial sonographic assessment of ventricular size in infants. PMID- 12557065 TI - Classification of venous malformations in children and implications for sclerotherapy. AB - OBJECTIVE: The purpose of this work is to present a simple and descriptive classification system for venous malformations (VMs) that may serve as a basis for interventional therapy, and to test its usefulness in a sample of consecutively referred paediatric patients. MATERIALS AND METHODS: The classification system we developed includes four types: type I, isolated malformation without peripheral drainage; type II, malformation that drains into normal veins; type III, malformation that drains into dilated veins; and type IV, malformation that represents dysplastic venous ectasia. The system was prospectively tested using phlebography in a sample of 43 children and adolescents with VMs who were referred for treatment during a 10-month period. Our hypothesis was that the type of VM would determine whether low-risk sclerotherapy was indicated. RESULTS: Thirteen (30%) patients had a type-I VM, 16 (37%) had a type-II, 9 (21%) had a type-III, and 5 (12%) had a type-IV malformation. In more than 90% of patients with a type-I or type-II lesion, sclerotherapy could be performed without any problems. In one third of patients with a type-III VM, sclerotherapy had to be withheld and one of nine (11%) developed a severe complication after therapy. Of the five patients with type-IV lesions, three (60%) had to be excluded from sclerotherapy. CONCLUSIONS: Our initial results indicate that sclerotherapeutic intervention in patients with type-III and type-IV VMs must be carefully considered, while it can be safely performed in low-risk patients with type-I and type-II lesions. PMID- 12557067 TI - Potential pitfalls in cranial sonography. AB - BACKGROUND: We present several anatomic variants of the brain and artifacts related to scanning techniques which could be misinterpreted as lesions on neonatal cranial sonography. MATERIALS AND METHODS: The findings were derived from US studies performed on 176 premature infants and 26 full-term newborns, using the anterior, posterior and mastoid fontanelles as acoustic windows. RESULTS: The pseudolesions are divided into three groups: ventricular system (asymmetric lateral ventricle size and coarctation of the lateral ventricles); choroid plexus ("split" choroid, "truncated" choroid and choroid cyst); and brain parenchyma (peritrigonal blush, thalamic pseudolesion, pseudo-absence of the inferior vermis, occipital pseudomass and calcar avis simulating intraventricular clot). We provide images of these pseudolesions and clues to their differentiation from true brain pathology. Images of several brain disorders are included for comparison. Knowledge of these potential pitfalls is essential for proper interpretation of US brain studies and will help to avoid the use of other more invasive diagnostic tests. CONCLUSIONS: Misleading images seen on US examination of the neonatal brain that could be misinterpreted as pathology are presented, with clues to their differentiation from true lesions. PMID- 12557068 TI - Fast-flow lingual vascular anomalies in the young patient: is imaging diagnostic? AB - AIM: To describe the imaging findings (MR imaging and angiography) of high-flow vascular anomalies of the tongue, hemangiomas and arteriovenous malformations (AVMs), with emphasis on the discrepant imaging findings in lingual AVMs. MATERIALS AND METHODS: Retrospective review of clinical records, histologic reports and imaging studies of five consecutive patients with high-flow lingual vascular anomalies. RESULTS: One patient had hemangioma (aged 1 month) and four patients had AVMs (aged 15 months, 6, 24, and 33 years). Diagnosis was made on the basis of histology in four lesions and was based on typical clinical history in one lesion. MR imaging and angiographic findings of the hemangioma were typical, but similar findings of focal hyperintense mass on T2-weighted images and angiographic stain were seen in three AVMs (patients aged 15 months, 6 and 33 years). On angiography, there was no nidus or direct arteriovenous (AV) shunting in one AVM (patient aged 15 months). The fourth AVM had typical MR imaging and angiographic findings. CONCLUSION: The imaging findings in lingual AVMs can be atypical or inconclusive and can mimic hemangiomas, especially in the young patient. Since treatment depends on accurate diagnosis, biopsy may be necessary for lesions with inconclusive imaging findings. PMID- 12557069 TI - The vascular "ring" sign in mesoblastic nephroma: report of two cases. AB - Mesoblastic nephroma (Bolande's tumour) is a rare renal tumour, but, overall, is the most frequent benign renal tumour in childhood and the most frequent renal tumour during the first year of life. It is increasingly diagnosed with US during the fetal period. We report the findings of two patients who showed the "ring" sign, an anechoic ring surrounding the tumour and typical of mesoblastic nephroma. We identified a Doppler signal in the ring in both cases, and pathology confirmed the vascular origin of this sign. PMID- 12557070 TI - Heterotopic gastric mucosa involving the gallbladder and biliary tree. AB - A case of heterotopic gastric mucosa in the common bile duct, cystic duct and gallbladder is reported in a 3-year-old girl with abdominal pain and jaundice. Abdominal US and CT showed dilatation of the biliary tree and a well-defined mass in the common bile duct that narrowed its lumen. The gallbladder was contracted in both examinations. The common bile duct and the gallbladder were resected and a choledochojejunostomy was performed. Although gastric heterotopy has been described throughout the entire length of the gastrointestinal tract, it is a very uncommon finding in the gallbladder and extremely rare in the biliary tree. A firm diagnosis of gastric heterotopia is based on the presence of fundal mucosa replete with parietal and chief cells. To our knowledge, this is the fifth reported case of heterotopic gastric tissue within the common bile duct, and the first to describe the US and CT findings. A relevant literature review and brief outline of the histological and radiological features are included in the discussion. PMID- 12557071 TI - Endoscopic internal biliary drainage in a child with malignant obstructive jaundice caused by neuroblastoma. AB - We describe a 13-year-old girl who underwent insertion of a Flexima biliary stent for obstructive jaundice due to compression of the extrahepatic bile duct by an enlarged lymph node secondary to neuroblastoma. This novel endoscopic internal biliary drainage procedure was safe and effective even for a child, and improved her quality of life. We further review other treatment options available for malignant obstructive jaundice in children. PMID- 12557072 TI - Orbital granulocytic sarcoma: an unusual presentation of acute myelocytic leukemia. AB - Granulocytic sarcoma is an unusual manifestation of acute myelogenous leukemia in children and presents a diagnostic dilemma when it precedes the development of systemic disease. We present CT and MRI findings of an extraconal mass proven to be granulocytic sarcoma in a 6-year-old otherwise healthy boy with several months' history of worsening unilateral proptosis. This case is unique in providing exquisite CT and MRI correlation and in demonstrating rapid response to therapy. Further, as cytogenetics were positive for the t(8,21) translocation, this case provides opportunity for discussion of the associated incidence of this translocation and concomitant better prognosis. PMID- 12557073 TI - Cavitary Rhodococcus equi pneumonia with endobronchial granulomas: report of an unusual case. AB - An unusual case of cavitary Rhodococcus equi pneumonia with endobronchial granulomas in congenital HIV infection is presented. The clinical features and radiological manifestations of pulmonary R. equi infection are discussed. PMID- 12557074 TI - Gastric duplication cyst: appearance on prenatal US and MRI. PMID- 12557075 TI - Persistent left superior vena cava in a preterm newborn. PMID- 12557077 TI - Attitudes of health care workers towards waking a terminally ill patient in the intensive care unit for treatment decisions. AB - OBJECTIVE: We examined whether health care workers would wake an intubated patient whose preferences are not known, and whether attitudes are influenced by how health care workers themselves would like to be treated if they were in the patient's place. DESIGN, SETTING, AND SUBJECTS: Convenience sample of 90 participants at a postgraduate lecture to anesthesiologists and related professions. Participants filled out questionnaires after a case presentation followed by two commentaries, one arguing against, the other for waking a 49-year old intubated patient suffering from a large, intratracheal, poorly differentiated metastatic squamous cell carcinoma of the lungs. The patient was not aware of the diagnosis and poor prognosis and had not expressed any preferences. RESULTS: Participants were almost equally divided between the two alternatives. Significant differences were found between professions concerning the willingness not to wake the patient (19.8% of nurses vs. 45% of physicians and others). There was a strong correlation between the preferences of the health care worker for her-/himself and what he/she would do if in charge of the patient. CONCLUSIONS: Our study shows that attitudes of health care workers towards waking and informing an intubated patient in the intensive care unit about a hopeless situation differ. Educational programs should ensure that physicians and nurses, especially when discussing and deciding withdrawal of vital support, are aware of theses differences and realize that their own behavior can be influenced by their own preferences if themselves in the patient's situation. PMID- 12557078 TI - Fatal hemorrhage due to an isolated dissection of the superior mesenteric artery. PMID- 12557079 TI - Manipulating afterload for the treatment of acute heart failure. A historical summary. PMID- 12557080 TI - [Bone substitutes]. AB - A large variety of different bone substitutes are available for the treatment of congenital or acquired bone defects as an alternative to bone transplantation. Complications associated with graft harvesting, limited donor resources, and the potential risk of transmission of infectious diseases have led to the development of multiple concepts of new bone substitutes to minimize the wellknown problems. This article intends to give an overview of the products currently available on the market. Inorganic materials such as ceramics from hydroxyapatite and/or tricalcium phosphate, calcium phosphate cements, calcium sulfates and bioglass,organic materials (polymers) and composites, xenografts,and glass ionomer cements are discussed. PMID- 12557081 TI - [Ceramic couplings in orthopedic surgery]. AB - Ceramic materials have been used as a coupling in total hip arthroplasty since the 1970s to solve the problem of polyethylene particle disease. Several problems with the material and the design have been identified and solved. Using inlays and ceramic heads of the latest generation offers the possibility of reducing the wear rate to as low as 0.001 mm per year. The problem of ceramic fractures is rare. Recently due to the manufacturing process some zirconia ceramic heads have been problematic. New developments with other ceramics are discussed. PMID- 12557082 TI - [Metal-on-metal pairs for hip prostheses]. AB - Due to engineering deficiencies as well as problems in the basic material, the first generation of metal-on-metal hip prostheses was not up to standard.Furthermore, unsatisfying clinical results led to a decrease in the use of these prostheses.Nevertheless, there were several cases which demonstrated good results in long-term clinical outcome. After distinct improvements in manufacturing and materials, metal-on-metal prostheses have made a come back. This second generation, which are known mainly under the names METASUL and SIKOMET,have been in clinical use for up to 10 years and the good results found in biomechanical set ups seem to be confirmed in clinical trials. However, a possible disadvantage of this second generation of metal-on-metal hip prostheses might be the production of large amounts of particulate wear debris in the nanometer size range. Whether the great number of small size particles with an extended metal surface and the possible distribution of these particles in the body have biological effects on the cells and tissues (such as allergic or toxic reactions) remains unknown. Among the tribological problems, there is currently a discussion on the possible inflammatory effect of these metal particles, which might play a crucial role in longterm, systemic reactions of the body to metal particles. PMID- 12557083 TI - [Polyethylene as an implant material]. AB - Since the early 1960s, ultrahigh-molecular weight polyethylene (UHMWPE) has been one of the most frequently and successfully used materials in total joint replacement. However, in the course of clinical usage, various weaknesses of the material became apparent including wear and aging properties. As a result of research on this polymer, several improvements in the quality of the raw material, the processing, and the sterilization have been introduced. These changes paired with an improved knowledge of the viscoelastic behavior of UHMWPE have led to an improved design of the components to meet the requirements of the given material characteristics and in turn to better clinical results. In the 1990s, investigations focused on the influence of radiation on the molecular structures of polyethylene. Gamma radiation treatment in air leads to an important decrease in the molecular weight with reduced tribological behavior and accelerated aging. In contrast, a nitrogen atmosphere during irradiation and storage can improve the linkage of the polyethylene and promote the positive effects of radiation treatment. This effect can be further intensified by increased radiation levels. Such highly cross-linked polyethylenes demonstrate extremely low wear rates in vitro;however, other material properties are also changed. First reports on fissures in highly cross-linked polyethylene explants demonstrate that only long-term clinical trials can allow a final verdict on the clinical potential of highly cross-linked polyethylenes. PMID- 12557084 TI - [Carbon fiber-reinforced plastics as implant materials]. AB - Carbon fiber-reinforced plastics have been used clinically as an implant material for different applications for over 20 years.A review of technical basics of the composite materials (carbon fibers and matrix systems), fields of application,advantages (e.g., postoperative visualization without distortion in computed and magnetic resonance tomography), and disadvantages with use as an implant material is given. The question of the biocompatibility of carbon fiber reinforced plastics is discussed on the basis of experimental and clinical studies. Selected implant systems made of carbon composite materials for treatments in orthopedic surgery such as joint replacement, tumor surgery, and spinal operations are presented and assessed. Present applications for carbon fiber reinforced plastics are seen in the field of spinal surgery, both as cages for interbody fusion and vertebral body replacement. PMID- 12557085 TI - [Bone cements based on polymethylmethacrylate]. AB - Bone cements based on polymethylmethacrylate (PMMA) remain an important material for anchorage of artificial joints. Polymers based on PMMA originally developed for dental surgery have been successfully used in bone surgery for more than 40 years. At first sight the cold-curing PMMA bone cement seems to be a rather simple material consisting of a powder and a liquid. But in fact it is a complex material fulfilling various functions at its application site after the implantation. Its properties vary according to the composition of its basic elements. They already play a decisive role for the working behavior during mixing of both components. The differences in the working behavior considerably affect the cementing technique and the accurate application in vivo. These influence the mechanical performance of the cured cement mantle and the long-term results of the implantation. Standardized test methods are used to characterize bone cements,whereas the clinical relevance of the test methods has to be evaluated critically.Additionally,PMMA bone cements act as a drug delivery system as a local carrier of antibiotics. This paper gives a review of the composition and properties of PMMA bone cements and their influence on practical application. PMID- 12557086 TI - [Surface modifications to improve biocompatibility and mechanical properties of orthopedic implants]. AB - State of the art surface modifications on metallic materials for orthopedic and dental implants permit clinical application if implant design,manufacturing process as well as function and duration of implantation are harmonized with each other. Keeping these prerequisites in mind,hydroxylapatite is suited for closer connection of fixation elements in hard tissue. PVD or PECVD modifications can make gliding surfaces or surfaces of fixation elements abrasion resistant if the pressure-area ratio does not exceed threshold values in cases of surfaces moving relative to each other PMID- 12557087 TI - [Allergic reactions to implant materials]. AB - The extent of the immune response upon implantation of metallic devices depends on the individual reactivity and on material characteristics. If specific T cellular sensitization occurs or an allergy to metal preexists, hypersensitive reactions to implant components may develop. They include eczema, impaired wound healing, and sterile osteomyelitis. The existence of allergy-induced implant loosening is still an open question. Further improvement of clinical allergological diagnostics, better understanding of peri-implantar immune reactions, and interdisciplinary collection of epidemiological data concerning allergy to implants will contribute to a better knowledge about tolerance of implant material in humans. PMID- 12557088 TI - [Integration of periosteum covered autogenous bone grafts with and without autologous chondrocytes. An animal experiment using the Gottinger minipig]. AB - Autologous osteochondral transplantation has the major disadvantage of significant damage to a healthy joint surface at the donor site. The purpose of this study was to examine the effect of autogenous chondrocytes injected into the periosteum of autologous bone grafts in order to provide an alternative method for cartilage repair. A total of 22 Gottinger minipigs were operated twice on both knees. The first operation served for cartilage biopsy for the chondrocyte culture. During the second operation an osteochondral defect was created in the medial facet of the trochlear groove. The defect was treated differently with an autologous cortico-cancellous bone cylinder,harvested from the proximal tibia.Group A: untreated defect (control);B: bone-graft;C: bone-graft covered with periosteum; D: bone-graft with periosteum and injected autologous chondrocytes. The animals were killed after 6, 12, 26 and 52 weeks. The regenerated areas were evaluated macroscopically, tested biomechanically (long term specimens; indentation-test) and a histological, blind evaluation was carried out according to a semi-quantitative scoring system. The periosteum covered bone cylinders in Groups C and D showed good repair of the bone and cartilage defect. The repaired tissue consisted predominantly of fibrocartilage with the partial formation of hyalin like tissue. The regenerated areas were integrated with the adjacent cartilage and were biomechanically superior when compared with the other groups. The additional injection of chondrocytes did not produce significantly better results. Our findings suggest that the transplantation of periosteum-covered bone cylinders may provide an alternative method for treating chondral and osteochondral defects and can be recommended for filling large donor site defects in joint surgery. The additional transplantation of chondrocytes does not seem to be justified. PMID- 12557089 TI - [Pathomorphology of parosteal osteosarcoma. Experience with 125 cases in the Hamburg Register of Bone Tumors]. AB - Parosteal osteosarcoma is a rare low-grade bone tumor. It occurs between the 2nd and 8th decade of life. In our own series of 125 parosteal osteosarcomas, most patients were aged between 30 and 40. The most frequent location was the distal dorsal femur. Until clearly proven otherwise, a bone-forming tumor in this location has to be regarded as a parosteal osteosarcoma. Of all parosteal osteosarcomas 77% are located in the lower extremity, with a female sex predominance in this location (62%); 18% are located in the upper extremity; 15% of all parosteal osteosarcomas are located in the humerus. Just 6% of all parosteal osteosarcomas are observed in the skull, the spine, and the pelvis. The tumor is characterized by hyalinized fibrous stroma with a low cell content without substantial nucleus polymorphism and variably dense bony trabeculae. The diagnosis can be difficult due to highly differentiated areas with fat tissue within the marrow and very uniformly bony structure. The operation material has to be analyzed very carefully, because the extent of dedifferentiated areas and most probably also the extent of invasion of the medullary cavity determine the prognosis and occurrence of recurrencies. The tumor is most commonly misinterpreted as osteochondroma or heterotopic ossification. Paying attention to all radiologic and histologic criteria,osteoblastic eccentrically located high grade osteosarcomas can be clearly distinguished from parosteal osteosarcoma. An intramedullary located low-grade osteosarcoma,which is differentiated like a parosteal osteosarcoma,is histologically indistinguishable from the parosteal variant. This variant requires the synthesis of all available data to find the correct diagnosis. The parosteal osteosarcoma shows like no other tumor the necessity of close cooperation of all involved disciplines for diagnosis and therapy. PMID- 12557090 TI - [Early detection of Parkinson syndrome]. PMID- 12557091 TI - [Procedure for reduction of allotransfusion in operative medicine]. PMID- 12557092 TI - [Hemangionar and vascular malformations. Classification, diagnostics and therapy]. PMID- 12557093 TI - [Teleteaching in otorhinolaryngology (part 1). Real-time transmission of a congress in picture and tone into the Internet]. AB - The transmission of scientific seminars into the Internet represents a new application of modern technologies for teleteaching. We digitally recorded 74 seminars and discussions during the 8th annual meeting of the German Society of Skull Base Surgery with five cameras and eight microphones, and transformed the data live with the real-Producer using a video card.This transferred data stream was sent via ISDN (128 kbit/s) to a real-server at the university, which was used for distribution in the net. The seminars could be called up in the display format of 320x240 pixels with a delay of 3 min (minimum: 30 s, as a function of the connection of the user) at each computer with Internet entrance. Altogether,we registered 3,284 accesses to the presentations following the congress until June 2002. The quality of picture transmission (24-36 kbit/s) and the quality of tone (audio stream: 5 kbit/s) were sufficient to understand the contents of the slide, video or computer presentation. The transmission of scientific meetings into the Internet can increase, with a justifiable technical expenditure, their efficiency by the ubiquitous availability of the information and the extension of the audience. PMID- 12557094 TI - [Hearing loss in a cultural-historical context. Part 1]. AB - BACKGROUND: It is rare to find anything written about hearing loss or its treatment in either representative art or in a medical-historical context in ear, nose and throat journals. METHODS: Examples dating back over the past 4000 years found in medical-historical, philosophical, and literary documents are presented and commented on. RESULTS: The ancient Egyptians and Greeks attempted to explain deafness and sought remedies,making use not only of empiric rational means but also magic and religion. Later, in the Middle Ages, examples from Christian iconography are found demonstrating miracle healing of deafness. Education of the deaf was not considered possible from the time of Aristotle to the sixteenth century, but from then on was organized on a large scale utilizing speech with gestures. PMID- 12557095 TI - [Operative voice pitch raising in male-to-female transsexuals. A survey of our technique and results]. AB - BACKGROUND: Transsexuality constitutes a complex,permanent transposition due to a paradoxical sense of belonging to the opposite sex. The persons concerned experience such a high degree of suffering that they strive to undergo operative sex reversal. Mainly in the case of male-to-female transsexuals this may often lead to a subsequent discrepancy between the external appearance and the character of the voice,since only a deepening of the voice can be achieved by hormonal treatment and not a raising of its pitch.Also,logopedic treatment on its own cannot,as a rule, guarantee a lasting success. In order to adapt the voice as an essential aspect of human identity to the external female appearance,a variety of phonosurgical operation techniques have been developed since the seventies. PATIENTS AND METHODS: In 1993 a modified cricothyroidal approximation via miniplates, following ISSHIKI's technique, has been performed at the ENT Department of the Martin-Luther-University of Halle-Wittenberg. This procedure is based upon an approximation of the cricoid and thyroid cartilages resulting in an increase in the vocal cord tension. This intervention does not affect the inner structures of the larynx and is on principle reversible. Combined with a chondrolaryngoplasty it allows for a reduction of the Adam's apple. So far 57 patients have undergone this surgical intervention. Laryngoscopy, stroboscopy and a detailed voice diagnostic, as well as ultrasound and computertomographic examinations of the larynx are performed pre and postoperatively in order to record the anatomical-physiological and functional data. RESULTS AND CONCLUSIONS: So far good functional results could be achieved showing an average rise of the medium voice range by approximately one fourth. As a long-term result a permanent raising of the basic voice pitch has been reported. The majority of the patients are satisfied with the functional and cosmetic effect reporting an enhanced self esteem. PMID- 12557096 TI - [Cricohyoidopexy (CHP) and Cricohyoidoepiglottopexy (CHEP). Indication, complications, functional und oncological results]. AB - BACKGROUND: Subtotal laryngectomy with Cricohyoido(epiglotto)pexy (CHEP and CHP) is a commonly used surgical procedure in France, Italy and North America, but it is rarely carried out in Germany,where most laryngeal carcinomas staged T1-T3 are resected endoscopically or with total laryngectomy. OBJECTIVE: To identify indications for the CHEP and CHP in a setting that uses endolaryngeal procedures as a standard approach to organ preserving surgery in laryngeal cancer patients. PATIENTS: Nineteen patients with primary (n=15) or recurrent (n=4), supra- or transglottic carcinoma or carcinoma of the anterior commissure staged (r)T1b-4N0 2cM0 were treated with subtotal laryngectomy with CHEP (with or without neck dissection/radiotherapy) between October 1997 and June 1999. RESULTS: Undisturbed deglutition without aspiration and respiration without tracheotomy was achieved in 17/19 patients. Three patients showed temporary pneumonia from aspiration and two patients needed further treatment for endolaryngeal synechia. Three patients died of unrelated causes. Four patients were diagnosed with local recurrence: Two of them died from tumor, two patients had curative total laryngectomy as salvage surgery. Fourteen patients are living free of disease 24-40 months after therapy. CONCLUSION: CHEP is a subtotal laryngectomy with increased postoperative morbidity, but good functional results. Assuming a scrupulous indication for the extended tumors the oncological results of the CHEP are satisfying, too. PMID- 12557097 TI - [Interstitial Nd:YAG laser therapy. Color-Doppler imaging (CDI)-guided laser therapy of hemangiomas and vascular malformations]. AB - BACKGROUND AND OBJECTIVE: Ultrasound-(US-) guided Nd:YAG laser therapy has been reported to be promising among different other options for treatment of hemangiomas and vascular malformations. The concept of CDI-guided interstitial laser therapy is therefore a further development based on the above mentioned therapeutic regime. PATIENTS AND METHODS: We examined 55 patients suffering from vascular lesions of different localization and extent pre-, intra-, and post operatively using color-Doppler imaging (CDI). All vascular lesions were classified due to their vascularization pattern. As accurate diagnosis was imperative we decided on possible laser treatment depending on the CDI classification. RESULTS: Intra-operative use of CDI rendered possible fiber guidance by sonographic navigation and therefore precise treatment of aberrant vessels and well-perfused tumor areas. In 34 cases (62%) we observed complete, in 21 cases (38%) partial regression of vascular lesions. CDI allowed post-operative proof of diminution of tumor vascularization in all cases. CONCLUSIONS: CDI guided interstitial Nd:YAG laser therapy is a minimally-invasive technique which allows replacement of conventional surgical procedures while achieving good cosmetic and functional results.CDI broadens the therapeutic possibilities in contrast to the former therapeutic concept of US-guided laser therapy due to pre , intra and post-operative utilization of this imaging technique while significantly diminishing the intra-operative risk for the patient. PMID- 12557098 TI - [Endoscopic assessment criteria in central laryngeal motility disturbances]. PMID- 12557099 TI - [Multimodal treatment strategy for Langerhans cell histiocytosis at head and neck manifestations]. AB - Langerhans cell histiocytosis (LCH) is a rare disorder with different clinical features. An established standardized treatment does not exist.We present a case report of a patient with localized LCH of the temporal bone and discuss the interdisciplinary treatment strategies. We reviewed the international literature and summarized the current knowledge. Beside a wait and see policy in cases without symptomatic disease, surgery, radiotherapy or chemotherapy, and combinations of these options are used as treatment modalities. While surgery or radiotherapy are preferred in localized (symptomatic) lesions, stage-adapted chemotherapeutic regimens are the treatments of choice in disseminated disease. Treatment selection depends on the individual clinical features. PMID- 12557100 TI - [Multifactorial dizziness]. AB - We report about a patient with recurrent dizziness, looked upon for about 8 years as Meniere's disease.He suffered from benign positional vertigo, which led to an additional psychogenic dizziness, connected to early live events. The different parts of complex dizziness, diagnosis and treatment will be described. PMID- 12557101 TI - [Cogan-I-syndrome. A rare differential diagnosis in progressive sensorineural hearing loss]. AB - We report on two patients affected by Cogan's syndrome since infancy.Cogan's syndrome is described as the association of abrupt or intermittent worsening of vestibulo-auditory function and non-syphilic interstitial keratitis or, in patients with "atypical" Cogan's syndrome, various severe, inflammatory eye diseases. The first patient was affected with different inflammatory eye diseases from her fifth year and became blind in one eye when she was 7 years old. At that time, a deterioration of her hearing ability was also diagnosed. The hearing loss progressed to a severe hearing disorder. The second patient suffered from different inflammatory eye diseases from his second year. When he was 3 years old,he was diagnosed with a severe hearing disorder that progressed.Cogan's syndrome was diagnosed when he was 13 years old. The recognition of Cogan's syndrome is particularly complex in young patients. The difficulty of the diagnosis requires a search for ophthalmic diseases in young children with acute and progressive vestibulo-auditory symptoms. PMID- 12557102 TI - [Current theories on the origin of cholesteatoma]. PMID- 12557103 TI - The comparison of reliabilities in dental imaging methods. AB - OBJECTIVES: Common practice in the statistical comparison of imaging instruments with limited reproducibility consists in the separate estimation of the instrument's reliabilities. However, as soon as one of the imaging methods is subject to item-specific bias (which has to be expected in many dentomaxillofacial imaging procedures), this approach will end in severe errors in reliability computation and in corresponding erroneous clinical conclusions. This paper seeks to point out these effects and to illustrate a more appropriate model for the comparison of instrumental reliabilities. METHODS: A standard reliability model was adjusted for item-specific bias and illustrated by the comparison of twice repeated planimetric cephalometry versus twice repeated noninvasive orthodontic video imaging (based on the Digigraph 100 device) in 50 children; the anterior cranial base length was used for illustration. RESULTS: The proposed model revealed pronounced inferiority of the video-based imaging system concerning its reliability compared with the X-ray based standard. Analysis using separate estimation of the two reliabilities would result in the reverse conclusion and thus falsely establish video imaging, which is in fact less reliable, as a superior diagnostic method. CONCLUSION: The reliabilities of dentomaxillofacial imaging methods have to be adjusted for potential item specific bias to avoid the erroneous conclusion of the superiority of a diagnostic innovation. PMID- 12557104 TI - On the in vitro biocompatibility of Elgiloy, a co-based alloy, compared to two titanium alloys. AB - AIM: The aim of the present investigation was to contribute to an understanding of the effects of surface topography and chemical composition on the corrosion behavior and thus the biocompatibility of Elgiloy (RMO, Denver, CO, USA), a common Co-based alloy. MATERIAL AND METHODS: The results are compared with those obtained for a binary NiTi alloy, Neo Sentalloy (GAC, Central Islip, NY, USA) and a beta-III-Ti alloy, TMA (Ormco, Glendora, CA, USA). In the present study, the surface topography and the chemical composition of two different grades of Elgiloy, Blue Elgiloy (soft) and Yellow Elgiloy (ductile), were examined by means of scanning electron microscopy (SEM) and energy-dispersive spectroscopy analysis (EDS). Their corrosion behavior in half-strength Ringer solution and in an artificial saliva solution according to Barrett [1] was investigated using potentiodynamic corrosion testing (PDC). The photometry-based PAN method was used to quantify the released Ni and Co ions. The in vitro biocompatibility of the two grades of Elgiloy was tested in three different cell cultures: in L929, a commercially available mouse fibroblast cell line, and in primary human epithelial cells and fibroblasts. RESULTS: The results of the corrosion testing showed satisfactorily high pitting corrosion potentials but lower repassivation potentials and a strong increase in current density once pitting had occurred. The photometric results revealed the release of Ni and Co ions in both tested electrolytes. The tested native surfaces exhibited numerous grinding and polishing grooves, inclusions and inhomogeneities of the microstructure. After corrosion testing the same surfaces displayed numerous signs of corrosion, especially in areas with microstructural inhomogeneities. In vitro biocompatibility testing showed a substantially reduced dehydrogenase activity in the presence of Elgiloy. The reduced quality of surface finish resulting from the manufacturing process led in the case of the tested Elgiloy types to decreased corrosion resistance with consequently reduced in vitro biocompatibility. CONCLUSIONS: In this context it is also conceivable that patients with a proven allergy to nickel, cobalt or chromium may react sensitively to the deployment of this alloy, at least in the surface quality tested by us. From this aspect, the introduction of a binding standard for the surface quality of materials used in orthodontic appliances is urgently recommended. PMID- 12557105 TI - Comparative cephalometric study of 10-year-old patients with complete unilateral cleft lip and palate. A cross-sectional study of the Hanover concept. AB - PATIENTS AND METHODS: The craniofacial morphology of 10-year-old male (n = 23) and female (n = 13) patients with unilateral cleft lip and palate was evaluated by means of cephalometric analysis. The control group comprised 40 non orthodontically treated non-cleft patients from the same population (20 boys, 20 girls) with neutroclusion, matched according to age and gender. Furthermore the results of the cephalometric analysis were compared with those of other cleft centers. Primary rehabilitation of all patients was performed according to a uniform concept at Hanover Medical School. All cephalometric radiographs were analyzed according to Ross [26]. This analysis permitted comparison with the results of two different cleft centers covering 107 patients, published by Ross. RESULTS: In comparison with the control group, both boys and girls with unilateral cleft lip and palate showed a significant retrusion and clockwise rotation of the mandible as well as a decreased vertical midfacial development. PMID- 12557106 TI - Results of transplanting developing third molars as part of orthodontics space management. Part 2: results following the orthodontic treatment of transplanted developing third molars in cases of aplasia and premature loss of teeth with atrophy of the alveolar process. AB - MATERIAL AND METHODS: The aim of this study was to assess the effects of atrophy of the alveolar process and subsequent orthodontic treatment on the transplantation results of developing third molars. The material consisted of 35 teeth transplanted to an atrophied alveolar process. 19 of these teeth were treated orthodontically subsequent to transplantation. The control group consisted of 61 developing molars transplanted to a new extraction socket. The transplanted teeth were followed up clinically and radiographically for a mean period of 3.2 years. RESULTS: The success rate was 85% for transplants to new extraction sockets and 84% for transplants to atrophied jaw sections with subsequent orthodontic treatment. In contrast, transplants to atrophied jaw sections without subsequent orthodontic treatment showed a significantly (p < or = 0.001) lower success rate of only 37.5%. These poorer results were due to persistent infraocclusion and ankylosis. CONCLUSIONS: Even in cases with atrophy of the alveolar process, a transplantation with subsequent orthodontic treatment represents a promising treatment concept, whereas teeth without subsequent orthodontic treatment showed a lower success rate. Orthodontic tooth movement had no negative effect on the healing rates of transplanted developing third molars. PMID- 12557107 TI - Transplantation of bicuspids in a case of aplasia of the lower central incisors. AB - The relatively unusual aplasia of both lower central incisors was treated by mesialization of the lateral incisors and cuspids, with a stable anterior dental arch thus being established. The resulting spaces in the posterior area were closed by autogenous transplantation of teeth 15 and 25 into the lower jaw. PMID- 12557108 TI - Transplantation of a lower bicuspid after traumatic loss of three upper incisors. AB - BACKGROUND: Prosthodontic and orthodontic procedures have been described for the treatment of patients with traumatic loss of teeth. PATIENT AND METHODS: Both upper central incisors and the upper right lateral incisor had to be extracted in an 8-year-old patient after failure of conservative procedures following a sports accident. The patient was treated on a temporary basis with a space retainer as a substitute for the teeth. Definitive therapy was carried out by transplanting a lower bicuspid germ into the upper arch with subsequent orthodontic gap closure. RESULTS: 8 years after the intervention, the transplanted tooth is still in place. The patient is fully rehabilitated both functionally and esthetically with no discomfort. CONCLUSIONS: The transplantation of bicuspid germs in conjunction with orthodontic gap closure is a valuable alternative to prosthodontic treatment, particularly in the case of loss or aplasia of several teeth. PMID- 12557109 TI - [Prevalence of type 1 diabetes-specific autoantibodies and of certain HLA patterns in celiac disease]. AB - BACKGROUND AND OBJECTIVE: An association between type 1 diabetes mellitus and celiac disease has been known for some time. One in ten type 1 diabetics have immunological markers for celiac disease (CD). But is there, conversely, an increased risk of CD for diabetics? This study was undertaken to answer this question by determining diabetes-associated antibodies and genetic factors in patients with a gluten-sensitive enteropathy (CD). PATIENTS AND METHODS: 68 patients with CD (48 females and 20 males) were investigated by determining the diabetes-associated serological marker GADA (glutamic acid decarboxylase antibodies). 1A-2A (insulinoma-associated protein 2 antibodies), ICA (Islet cell antibodies) and IAA (insulin autoantibodies). Among this cohort were 60 patients up to the age of 25 years and eight adults (average age 41.7 years). In 36 of these patients the HLA was also determined. RESULTS: GADA was found in 6 patients (9%), 1A-2A in eight (12%) and IAA in 21. ICA were not demonstrated in any. Five of the CD patients were positive for several markers. One child, positive for autoantibodies already had manifest diabetes at the time of investigation. None of the patients with autoantibodies had an abnormal glucose metabolism one year later. HLA-DR3, that occurs in both CD and diabetes, was demonstrated in 78% of the patients with CD. The most common constellation, HLA-DR3-DQ2/HLA-DR7-DQ2, was found in 31%. CONCLUSION: This investigation indicates a genetic association between celiac disease and diabetes. Nonetheless, the risk of developing diabetes mellitus is only minimally higher in patients with CD than in the normal population. Therefore, general screening cannot be recommended at present. Further studies will be needed. PMID- 12557110 TI - [Sildenafil does not change coronary flow reserve in diabetics with erectile dysfunction]. AB - BACKGROUND AND AIM OF STUDY: Disturbance of the microvascular coronary circuit is common in diabetics with erectile dysfunction. We investigated effects of sildenafil on coronary flow reserve (CFR) of the left anterior descending branch. PATIENTS AND METHODS: 43 diabetics (aged 59 +/- 7 years) with erectile dysfunction and without symptoms of coronary artery disease were selected. Cardiac diagnosis, including stress ECG and echocardiography was performed in all. Because of the clinical suspicion of coronary artery disease coronary angiography was performed in 16 of them. Severe coronary artery disease was confirmed in 12 patients who were excluded from further analyses as well as 10 diabetics in whom coronary flow measurements were not possible. In the other 21 diabetics, adenosine-mediated CFR was calculated at baseline and 1 hour after ingestion of 50 mg sildenafil by transthoracic Doppler echocardiography. RESULTS: CFR at baseline was at the lower level of the normal range in 17/21 diabetics (median 245 %, range 210 - 490 %). CFR decreased insignificantly in 12/21 patients after sildenafil administration (Delta CFR -10 %, p = 0.3). Patients with a body mass index > 25 kg/m(2), and left ventricular hypertrophy had the highest reduction of CFR after sildenafil, but a drop of the CFR below 200 % was not observed in any patient. Systemic blood pressure dropped significantly from 130/80 mmHg to 120/72 mmHg (p < 0.002). CONCLUSION: Diabetics with erectile dysfunction often have a CFR in the lower range of normal. Sildenafil did not further reduce CFR. Asymptomatic, severe coronary artery disease often can be found in diabetics with erectile dysfunction. Cardiological screening for contraindications for sildenafil seems mandatory in diabetics with a high cardiovascular risk profile. PMID- 12557111 TI - [Brain infarctions caused by paradoxical embolism during long-distance flights]. AB - HISTORY AND ADMISSION FINDINGS: Over the period of one year three long-distance airline passengers (aged 21, 63, and 64 years) were admitted to our department because of a first-time acute neurological deficit having occurred during their long-distance flights. INVESTIGATIONS: In all three cases acute stroke MRI showed embolic cerebral ischemia, and transesophageal echocardiography revealed a persistent foramen ovale (PFO). Venous duplex and compression ultrasonography of the legs showed no signs of thrombosis. Extra- and transcranial Doppler, Holter ECG recording, routine blood analysis and additional tests for thrombophilia (incl. lupus anticoagulans, APC resistance, protein C, S, and AT III) revealed no signs of cardiovascular disease or other stroke causes. Only in case 3 the prothrombin gene G 20210A was found. DIAGNOSIS, TREATMENT AND COURSE: Ischemic stroke due to paradoxical embolism through a PFO was diagnosed in all three patients. One patient recovered fully within 2 days, one was discharged with a persistent motor deficit and the third patient died subsequently as a result of multiple cerebral infarctions accompanied by massive pulmonary embolism. CONCLUSION: These three cases illustrate that paradoxical embolic stroke is a possible severe complication of long-distance air travel in passengers with a PFO and this should be taken into account when deciding upon individual risk-adjusted prophylactic measures. PMID- 12557112 TI - [Evidence-based treatment of methotrexate-resistant rheumatoid arthritis. Cytokine antagonists]. PMID- 12557113 TI - [Specific immunotherapy in allergic asthma]. PMID- 12557114 TI - [Images of man and distributive justice in health care. Ethical considerations in theological perspective]. PMID- 12557115 TI - [Old or new antihypertensive drugs? Consequences from ALLHAT. Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial]. PMID- 12557116 TI - [Use of varicella zoster live vaccine in older patients]. PMID- 12557117 TI - [Could a central facial paralysis become spastic?]. PMID- 12557118 TI - [When is the anaesthesia "deep" enough?]. PMID- 12557119 TI - [Intraoperative awareness]. AB - The first cases of general anesthesia were already cases with awareness. Until today, case reports of patients with awareness are published. These published cases are likely to be the top of the iceberg, as most patients with postoperative recall do not inform their anesthesiologist. Incidence of awareness with recall is between 0.1 and 0.2 %. In a large multicenter-study, incidence of recall was 0.1 % without, and 0.18 % with the use of muscle relaxants. The risk is increased with decreased doses of anesthetics, e.g. in patients with hemodynamic instability (trauma cases), patients undergoing cesarean section or cardiac surgery. Intraoperative awareness does not necessarily cause explicit (conscious) memory. Even in the absence of explicit memory, implicit (unconscious) memory can still have consequences for the patient. In the worst case, it can cause post-traumatic stress disorder. There is doubt whether patients may profit from positive suggestions given during intraoperative awareness. Recommendations to administer benzodiazepines to prevent explicit memory must be reconsidered. Complete neuromuscular block should be avoided whenever possible. If a patient is thought to be aware, he should be contacted, his situation should be explained and affirming comments should be given until consciousness is lost again. Postoperative visit should include questions about awareness and recall. The anesthetist should not disbelieve reported recall. Explanation of what had happened and referral to an experienced psychologist must be offered. Thus, the incidence of severe sequelae should decrease. PMID- 12557120 TI - [Patient's emotional satisfaction with anaesthetic care]. AB - OBJECTIVE: The evaluation of services by patients is an integral part of continuous quality improvement in anaesthesia. Patient satisfaction is affected by various objective and subjective variables. Individual experiences, impressions and needs represent an important part of overall patient satisfaction. METHODS: Data of 519 patients were collected in a standardised anonymous questionnaire independent of the post-anaesthetic visit. RESULTS: Anaesthesia staff appeared to be friendly to 97 % of patients asked, 95 % of patients wanted to be treated by the same staff in case of another anaesthesia, nearly 87 % felt relaxed and satisfied after anaesthesia. Up to 26 % of patients felt tense after pre-anaesthetic visit and 38 % before anaesthesia, 35 % felt that waiting before operation was endless and 20 % complained about loss of humaneness because of technical equipment and monitoring. Significant differences could be found between specific groups of patients: Older and male patients were in all more satisfied compared to younger and female patients, patients after regional anaesthesia were more satisfied compared to ones after general anaesthesia. CONCLUSION: The inquiry of subjective patient impressions can be performed in anaesthesia departments without much effort. The results help to identify and avoid psycho-vegetative stressing situations and to improve patient satisfaction with anaesthesia. PMID- 12557121 TI - [Comparison of different laryngeal mask airways in a resuscitation model]. AB - OBJECTIVE: The standard laryngeal mask airway LMA-Classic is recommended in the ILCOR guidelines as alternative to facemask and tracheal tube during cardiopulmonary resuscitation. LMA-Unique, LMA-Fastrach and LMA-ProSeal are additional variants that are compared with the standard LMA in a resuscitation model. METHODS: Tidal volumes, chest compressions and signs of gastric inflation are measured in a standardized resuscitation model (Ambu Cardiac Care Trainer with notebook and thumper). Ten 3-minute resuscitation cycles were performed with facemask and all LMAs (Classic, Unique, Fastrach, ProSeal, all size 4) with a ventilation : compression ratio of 2:15. To allow comparison with tracheal tube and to judge safety margins, another ten resuscitation cycles were performed with tracheal tube and all laryngeal masks with continuous chest compressions after two initial ventilations. The bag-valve device used for ventilation was replaced by an automatic transport ventilator in a third series with continuous chest compressions. Cuff pressures were set at 80 cm H2O. RESULTS: During interrupted chest compressions, adequate ventilation was possible with all devices. Tidal volumes for facemask, LMA-Fastrach and LMA-ProSeal were significantly (p < 0.001) higher than with LMA- Classic and LMA-Unique. During continuous chest compressions, significant differences (p < 0.001) occurred when comparing tracheal tube, Fastrach and ProSeal with the other two laryngeal mask airways, which did not reach recommended tidal volumes. During ventilation with the automatic transport ventilator, values for all devices except LMA-ProSeal dropped significantly, adequate ventilation was possible with tracheal tube, Fastrach and ProSeal. Signs of gastric inflation were found during ventilation with facemask and - to a lesser extent - with LMA-Classic. CONCLUSION: In the resuscitation model chosen, all laryngeal mask airways are possible alternatives for ventilation during cardiopulmonary resuscitation. The new LMA-ProSeal and also the LMA-Fastrach allow higher tidal volumes even during elevated intrathoracic pressures caused by continuous chest compressions, performing superior to the standard laryngeal mask airway. For inclusion of these devices in the ILCOR guidelines, further research in patients is warranted. PMID- 12557122 TI - [Acute myocardial infarction after blunt polytrauma -- successful coronary intervention]. AB - Acute myocardial infarction following blunt chest trauma is a well reported but rare finding. Especially in severely injured patients the optimal therapy of the myocardial infarction is not well established, since anticoagulants, platelet aggregation inhibitors or thrombolytics are frequently contraindicated under these conditions. We report a case of a 41-year-old man, who presented with an acute myocardial infarction in combination with a severe polytrauma (multiple rib fractures, hematothorax, pelvic bone fractures, multiple injuries of intestinal organs) after a motorcycle accident with a blunt chest and abdominal trauma. After surgical treatment of the injuries of the bones and the intestinal organs a coronary angiography was immediately performed. The left anterior descending and the circumflex coronary artery were occluded in the mid-portion of the vessels. Coronary recanalization by PTCA and the implantation of coronary stents were successful in both vessels. Despite of a non-optimal blood flow after recanalization and stenting in one vessel (LAD TIMI II flow after recanalization), and a non-optimal accompanying medical therapy, during and after intervention (intravenous heparin starting 8 hours after the coronary intervention and platelet inhibitors starting 4 days after the intervention) the coronary angiogram after 2 months documented both vessels patent without a reocclusion or a restenosis. The case report documents, that in traumatic myocardial infarctions the treating of both, the attending injuries and the myocardial ischemia, is feasible. Early coronary angiography and coronary interventions, with or without stent-implantation, are indicated, even in cases in which an adequate accompanying medical therapy with heparin and platelet inhibitors is contraindicated. PMID- 12557123 TI - [Standards in the diagnosis and treatment of sepsis]. PMID- 12557124 TI - Human population genetic structure and inference of group membership. AB - A major goal of biomedical research is to develop the capability to provide highly personalized health care. To do so, it is necessary to understand the distribution of interindividual genetic variation at loci underlying physical characteristics, disease susceptibility, and response to treatment. Variation at these loci commonly exhibits geographic structuring and may contribute to phenotypic differences between groups. Thus, in some situations, it may be important to consider these groups separately. Membership in these groups is commonly inferred by use of a proxy such as place-of-origin or ethnic affiliation. These inferences are frequently weakened, however, by use of surrogates, such as skin color, for these proxies, the distribution of which bears little resemblance to the distribution of neutral genetic variation. Consequently, it has become increasingly controversial whether proxies are sufficient and accurate representations of groups inferred from neutral genetic variation. This raises three questions: how many data are required to identify population structure at a meaningful level of resolution, to what level can population structure be resolved, and do some proxies represent population structure accurately? We assayed 100 Alu insertion polymorphisms in a heterogeneous collection of approximately 565 individuals, approximately 200 of whom were also typed for 60 microsatellites. Stripped of identifying information, correct assignment to the continent of origin (Africa, Asia, or Europe) with a mean accuracy of at least 90% required a minimum of 60 Alu markers or microsatellites and reached 99%-100% when >/=100 loci were used. Less accurate assignment (87%) to the appropriate genetic cluster was possible for a historically admixed sample from southern India. These results set a minimum for the number of markers that must be tested to make strong inferences about detecting population structure among Old World populations under ideal experimental conditions. We note that, whereas some proxies correspond crudely, if at all, to population structure, the heuristic value of others is much higher. This suggests that a more flexible framework is needed for making inferences about population structure and the utility of proxies. PMID- 12557125 TI - The constitutional t(17;22): another translocation mediated by palindromic AT rich repeats. AB - We have demonstrated that the breakpoints of the constitutional t(11;22) are located at palindromic AT-rich repeats (PATRRs) on 11q23 and 22q11. As a mechanism for this recurrent translocation, we proposed that the PATRR forms a cruciform structure that induces the genomic instability leading to the rearrangement. A patient with neurofibromatosis type 1 (NF1) had previously been found to have a constitutional t(17;22) disrupting the NF1 gene on 17q11. We have localized the breakpoint on 22q11 within the 22q11-specific low-copy repeat where the breakpoints of the constitutional t(11;22)s reside, implying a similar palindrome-mediated mechanism for generation of the t(17;22). The NF1 gene contains a 195-bp PATRR within intron 31. We have isolated the junction fragments from both the der(17) and the der(22). The breakpoint on 17q11 is close to the center of the PATRR. A published breakpoint of an additional NF1-afflicted patient with a constitutional t(17;22) is also located close to the center of the same PATRR. Our data lend additional support to the hypothesis that PATRR mediated genomic instability can lead to a variety of translocations. PMID- 12557129 TI - Acute gastroenteritis outbreaks on cruise ships linked to Norwalk-like viruses. PMID- 12557126 TI - HLA-DR2 dose effect on susceptibility to multiple sclerosis and influence on disease course. AB - Models of disease susceptibility in multiple sclerosis (MS) often assume a dominant action for the HLA-DRB1*1501 allele and its associated haplotype (DRB1*1501-DQB1*0602 or DR2). A robust and phenotypically well-characterized MS data set was used to explore this model in more detail. A dose effect of HLA-DR2 haplotypes on MS susceptibility was revealed. This observation suggests that, in addition to the role of HLA-DR2 in MS, two copies of a susceptibility haplotype further increase disease risk. Second, we report that DR2 haplotypes modify disease expression. There is a paucity of benign MS and an increase of severe MS in individuals homozygous for DR2. Concepts of the molecular mechanisms that underlie linkage and association of the human leukocyte antigen (HLA) region to MS need to be revised to accommodate these data. PMID- 12557130 TI - National gnotobiotic rodent resource. PMID- 12557131 TI - Impact factors--a good thing? PMID- 12557132 TI - Image of the month. Sweet's syndrome. PMID- 12557133 TI - Serious lower gastrointestinal clinical events with nonselective NSAID or coxib use. AB - BACKGROUND & AIMS: Epidemiologic studies suggest nonsteroidal anti-inflammatory drugs (NSAIDs) increase the risk for lower gastrointestinal (GI) clinical events, but data from prospective trials are lacking. Cyclooxygenase (COX)-2-selective inhibitors decrease upper GI clinical events but the effect on lower GI events has not been determined. We performed a post hoc analysis of serious lower GI clinical events with a nonselective NSAID and a COX-2-selective agent in a prospective, double-blind, randomized GI outcomes trial. METHODS: A total of 8076 rheumatoid arthritis patients 50 years or older (or 40 years or older on corticosteroid therapy) expected to require NSAIDs for 1 year or greater were randomly assigned to naproxen 500 mg twice daily or rofecoxib 50 mg daily. The rate of serious lower GI clinical events, defined as bleeding with a 2 g/dL drop in hemoglobin or hospitalization, or hospitalization for perforation, obstruction, ulceration, or diverticulitis, was determined. RESULTS: The rate of serious lower GI events per 100 patient-years was 0.41 for rofecoxib and 0.89 for naproxen (relative risk, 0.46; 95% confidence interval [CI], 0.22-0.93; P = 0.032). Serious lower GI events accounted for 39.4% of all serious GI events (complicated upper GI event or lower GI event) among patients taking naproxen and 42.7% among those taking rofecoxib. CONCLUSIONS: Serious lower GI events occurred at a rate of 0.9% per year in rheumatoid arthritis patients taking the nonselective NSAID naproxen, accounting for nearly 40% of the serious GI events that developed in these patients. Serious lower GI events were 54% lower with the use of the selective COX-2 inhibitor rofecoxib. PMID- 12557135 TI - Low-dose growth hormone in adult home parenteral nutrition-dependent short bowel syndrome patients: a positive study. AB - BACKGROUND & AIMS: Controversy surrounds a 3-week treatment with a high-dose (0.14 mg. kg(-1). day(-1)) growth hormone (GH), glutamine, and high carbohydrate diet in home parenteral nutrition (HPN)-dependent patients with short-bowel syndrome (SBS). This study assessed treatment with low-dose GH in these patients. METHODS: Twelve adult HPN-dependent (duration, 7 +/- 1 years; mean +/- SEM) patients with SBS (small-bowel remnant length, 48 +/- 11 cm) who were on an unrestricted hyperphagic diet were randomized in a double-blind, placebo controlled, crossover study. Patients received daily low-dose GH (0.05 mg. kg( 1). day(-1)) and placebo for two 3-week periods separated by a 1-week washout period. Net intestinal absorption of macronutrients was assessed using a duplicate diet; nutritional assessment and blood tests were performed. Data from each group were compared using Wilcoxon rank sum test. RESULTS: Treatment with GH increased intestinal absorption of energy (15% +/- 5%, P < 0.002), nitrogen (14% +/- 6%, P < 0.04), carbohydrates (10% +/- 4%, P < 0.04), and fat (12% +/- 8%, NS). The increased food absorption represented 37% +/- 16% of total parenteral energy delivery. Body weight (P < 0.003), lean body mass (P < 0.006), D-xylose absorption (P < 0.02), insulin-like growth factor 1 (P < 0.002), and insulin-like growth factor binding protein 3 (P < 0.002) increased, whereas uptake of GH binding protein decreased (P < 0.01), without any major adverse effect. CONCLUSIONS: Three weeks of low-dose GH significantly improved intestinal absorption in HPN-dependent SBS patients who were on a hyperphagic western diet. PMID- 12557136 TI - The cost-effectiveness of psychotherapy and paroxetine for severe irritable bowel syndrome. AB - BACKGROUND & AIMS: Psychotherapy and antidepressants are effective in patients with severe irritable bowel syndrome (IBS), but the cost-effectiveness of either treatment in routine practice has not been established. METHODS: Patients with severe IBS were randomly allocated to receive 8 sessions of individual psychotherapy, 20 mg daily of the specific serotonin reuptake inhibitor (SSRI) antidepressant, paroxetine, or routine care by a gastroenterologist and general practitioner. Primary outcome measures of abdominal pain, health-related quality of life, and health care costs were determined after 3 months of treatment and 1 year later. RESULTS: A total of 257 subjects (81% response rate) from 7 hospitals were recruited; 59 of 85 patients (69%) randomized to psychotherapy and 43 of 86 (50%) of the paroxetine group completed the full course of treatment. Both psychotherapy and paroxetine were superior to treatment as usual in improving the physical aspects of health-related quality of life (SF-36 physical component score improvement, 5.2 [SEM, 1.26], 5.8 [SEM, 1.0], and -0.3 [SEM, 1.17]; P < 0.001), but there was no difference in the psychological component. During the follow-up year, psychotherapy but not paroxetine was associated with a significant reduction in health care costs compared with treatment as usual (psychotherapy, $976 [SD, $984]; paroxetine, $1252 [SD, $1616]; and treatment as usual, $1663 [SD, $3177]). CONCLUSIONS: For patients with severe IBS, both psychotherapy and paroxetine improve health-related quality of life at no additional cost. PMID- 12557137 TI - Hepatitis C, iron status, and disease severity: relationship with HFE mutations. AB - BACKGROUND & AIMS: Mild to moderate hepatic iron loading is common in patients with chronic hepatitis C. We sought to determine whether mutations in the hemochromatosis gene, HFE, are associated with iron overload and acceleration of disease progression in hepatitis C patients. METHODS: A total of 316 patients with chronic hepatitis C were studied: 198 consecutive patients undergoing liver biopsy for compensated liver disease and 118 who underwent liver transplantation for end-stage liver disease. Serum iron studies, quantitative hepatic iron concentration, histologic activity index, and HFE genotype were determined. RESULTS: Among patients with compensated liver disease, the presence of HFE mutations was independently associated with elevations in serum iron level, serum transferrin-iron saturation, serum ferritin level, and hepatic iron index (P < 0.05). After adjustment for duration of infection with hepatitis C virus, HFE mutations were also independently associated with the presence of bridging fibrosis or cirrhosis (odds ratio, 18; 95% confidence interval, 1.7-193). HFE mutations were not independently associated with iron loading in patients with end-stage liver disease. There was no significant difference in the prevalence of HFE mutations between patients with compensated and end-stage liver disease (42% vs. 33%, respectively; P = 0.67). CONCLUSIONS: The presence of HFE mutations is independently associated with iron loading and advanced fibrosis in patients with compensated liver disease from chronic hepatitis C, especially after controlling for duration of disease. These results suggest that HFE mutations accelerate hepatic fibrosis in hepatitis C but may not be responsible for progression to end stage liver disease. PMID- 12557138 TI - Basal core promoter mutations of hepatitis B virus increase the risk of hepatocellular carcinoma in hepatitis B carriers. AB - BACKGROUND & AIMS: Hepatitis B viral (HBV) genotype C is associated with the development of hepatocellular carcinoma (HCC) compared with genotype B; however, the virologic factors contributing to the pathogenic differences remain unknown. We investigated the prevalence of T1762/A1764 basal core promoter mutant in a cohort of 250 genotype B- or C-infected HBV carriers with different stages of liver disease to clarify a possible role for this mutant in hepatocarcinogenesis. METHODS: The sequences of basal core promoter of HBV genome were determined in 60 inactive HBV carriers and 190 patients with histologically verified chronic liver disease and HCC. RESULTS: Genotype C has a higher prevalence of T1762/A1764 mutation than genotype B (odds ratio, 5.18; 95% confidence interval [CI], 2.59 10.37; P < 0.001). The likelihood of T1762/A1764 mutation parallels the progression of liver disease, from 3% in inactive carriers to 64% in HCC patients (odds ratio, 20.04; 95% CI, 7.25-55.41; P < 0.001). By multiple logistic regression analysis, patients with T1762/A1764 mutation were significantly associated with the development of HCC than those without (odds ratio, 10.60; 95% CI, 4.92-22.86; P < 0.001), and the risk was observed for both genotypes B and C. In addition, the prevalence of T1762/A1764 mutation in younger HCC patients was comparable with older HCC patients but was significantly higher than that in age matched inactive carriers, irrespective of genotypes. CONCLUSIONS: Our data suggest that HBV carriers with T1762/A1764 basal core promoter mutant are at increased risk for HCC and that this mutant may contribute to the pathogenesis of HBV infection. PMID- 12557139 TI - Defective cellular localization of mutant ATP7B in Wilson's disease patients and hepatoma cell lines. AB - BACKGROUND & AIMS: Wilson's disease, a hereditary disorder caused by mutations in the Wilson's disease gene (ATP7B), leads to hepatic and/or neurological pathology resulting from cellular copper overload. In vitro studies showed that ATP7B, located in the trans-Golgi network, traffics to a cytoplasmic vesicular compartment in response to increased copper concentration. Mislocalization and failed intracellular trafficking of ATP7B mutants are suggested to be among disease-causing mechanisms; however, the effect of mutations on ATP7B localization in human tissues has not been directly shown. Therefore, we characterized the subcellular localization of normal and mutant ATP7B in human livers and in hepatoma cell lines. METHODS: Subcellular distribution of ATP7B in liver tissue from 3 control individuals and 3 Wilson's disease patients harboring a homozygous H1069Q-ATP7B mutation was analyzed by using immunogold electron microscopy. In addition, 14 ATP7B mutants tagged to green fluorescent protein were generated and expressed in HuH-7 and HepG2 cells; intracellular localization of these mutants was characterized by confocal microscopy. RESULTS: In hepatocytes, ATP7B was localized in trans-Golgi vesicles, whereas H1069Q-ATP7B was trapped in the endoplasmic reticulum. Similar results were observed for wild type ATP7B and H1069Q-ATP7B expressed in hepatoma cells. Most ATP7B proteins harboring missense mutations were distributed similarly to wild-type ATP7B. In contrast, truncated ATP7B mutants showed a diffuse, clustered, cytoplasmic pattern, distinct from the trans-Golgi network or endoplasmic reticulum. CONCLUSIONS: These results provide a detailed demonstration of the ATP7B distribution in control and diseased human livers and indicate that several Wilson's disease mutations lead to incorrect localization of ATP7B to distinct cell compartments. PMID- 12557140 TI - Liver transplantation improves hepatic myelopathy: evidence by three cases. AB - BACKGROUND & AIMS: Hepatic myelopathy is a rare complication of chronic liver disease, causing progressive spastic paraparesis. Today, no therapy of this disorder has been established. Commonly used therapeutic strategies for hepatic encephalopathy aiming at the reduction of plasma ammonia levels such as protein restriction, oral neomycin, lactulose, or ornithine aspartate fail to improve the symptoms of hepatic myelopathy. The aim of this study was to find out whether orthotopic liver transplantation (OLT) may improve hepatic myelopathy. METHODS: Follow-up examinations of 3 patients with severe hepatic myelopathy before and after OLT. RESULTS: In all 3 patients, the neurologic status improved significantly after liver transplantation. The grade of improvement was related to the time interval between onset of the first symptoms of hepatic myelopathy and liver transplantation. CONCLUSIONS: Early recognition of hepatic myelopathy is important because timely liver transplantation as an established therapy for end-stage liver disease offers the chance of complete recovery from hepatic paraparesis. PMID- 12557141 TI - Associations of chemokine system polymorphisms with clinical outcomes and treatment responses of chronic hepatitis C. AB - BACKGROUND & AIMS: CCR5Delta32, a 32-base pair deletion of the CC chemokine receptor (CCR) 5 gene, is associated with slowed human immunodeficiency virus disease progression in heterozygotes and protection against infection in homozygotes. A recent study found a higher than expected frequency of CCR5Delta32/Delta32 in patients with hepatitis C virus infection. The roles of other disease-associated chemokine system polymorphisms have not been evaluated in hepatitis C virus infection. METHODS: Six chemokine system polymorphisms (CCR5Delta32, CCR5 promoter 59029-G/A, CCR2 -64I, RANTES [regulated upon activation, normal T cells expressed and secreted] -403 -G/A, and -28 -C/G and stromal derived factor 1 -3'A) were studied in 417 patients with liver diseases (339 with hepatitis C) and 2380 blood donors. The clinical parameters of hepatitis C virus infection were compared between carriers and noncarriers of each genetic variant. RESULTS: The frequency of CCR5Delta32 homozygosity was 0.8% in whites with hepatitis C virus and 1.1% in controls (P = 0.75). The CCR5Delta32 allele was not associated with any of the clinical parameters of hepatitis C virus infection. Hepatitis C virus-seropositive whites with the RANTES -403-A allele were less likely to have severe hepatic inflammation compared with those without (odds ratio, 0.34; P = 0.03). In multivariate analysis, the CCR5 promoter 59029 -A allele was marginally associated with a sustained response to interferon therapy (odds ratio, 3.07; P = 0.048). CONCLUSIONS: In this cohort, the frequency of CCR5Delta32 homozygosity in patients with hepatitis C was similar to controls. The high prevalence of CCR5Delta32 homozygosity in the hepatitis C virus patients of the earlier study likely reflects resistance to human immunodeficiency virus infection in hemophiliacs rather than a susceptibility to hepatitis C virus infection. Expression of CCR5 and RANTES may be important in the modulation of hepatic inflammation and response to interferon therapy in chronic hepatitis C. PMID- 12557142 TI - Peroxisome proliferator-activated receptor gamma ligands suppress colon carcinogenesis induced by azoxymethane in mice. AB - BACKGROUND & AIMS: Peroxisome proliferator-activated receptor gamma (PPARgamma) is known to regulate growth arrest and terminal differentiation of adipocytes and is used clinically as a new class of antidiabetic drugs. Recently, several studies have reported that treatment of cancer cells with PPARgamma ligands induces cell differentiation and apoptosis, suggesting a potential application as chemopreventive agents against carcinogenesis. However, contradictory results have been reported with regards to the biologic role of PPARgamma in carcinogenesis. Tanaka et al.(24) have recently reported the suppressive effect of a PPARgamma ligand, troglitazone (Tro), on the formation of aberrant crypt foci (ACF) in rats. In the present study, 3 different kinds of PPARgamma ligands were subjected to the experiments to confirm their suppressive effects on colon carcinogenesis. METHODS: Three PPARgamma ligands, pioglitazone (Pio) (200 ppm), rosiglitazone (Rosi) (200 ppm), and Tro (1000 ppm) were investigated on the induction of ACF, a putative precancerous lesion of the colon, and colon tumor formation using an azoxymethane (AOM)-induced colon cancer model in BALB/c mice, and dose dependency of a PPARgamma ligand was also examined. RESULTS: PPARgamma ligands reduced the ACF formation by AOM (10 mg/kg body weight) and induction of colon tumors were also markedly suppressed by a continuous feeding of Pio at 200 ppm. CONCLUSIONS: Our findings indicate that PPARgamma ligands are indeed potential chemopreventive agents for colon carcinogenesis. PMID- 12557143 TI - Effects of HIV-1 Tat protein on ion secretion and on cell proliferation in human intestinal epithelial cells. AB - BACKGROUND & AIMS: Severe diarrhea and enteropathy of unknown origin are frequent in patients infected with human immunodeficiency type 1 virus (HIV-1). The HIV-1 transactivating factor protein (Tat) is a key factor in the pathogenesis of acquired immunodeficiency syndrome. We investigated whether Tat could directly induce ion secretion and cell damage in enterocytes. METHODS: Electrical parameters (ion transport studies) were measured in Caco-2 cell monolayers and in human colonic mucosa specimens mounted in Ussing chambers. The effect of Tat on intestinal mucosa integrity was determined by monitoring the transepithelial electrical resistance of Caco-2 cell monolayers. (3)H-thymidine incorporation and cell count were used to evaluate the effect of Tat on cell growth. Intracellular calcium concentrations were measured at the single-cell level using microfluorometry technique. RESULTS: Tat protein induced ion secretion in Caco-2 cells and in human colonic mucosa similar to that induced by bacterial enterotoxins. It also significantly prevented enterocyte proliferation. In both instances, the effect of Tat was maximum at concentrations within the range detected in the sera of HIV-1-infected patients. Anti-Tat antibodies inhibited both effects. Ion secretion and the antiproliferative effects were mediated by L type Ca(2+) channels. An increase in intracellular calcium concentration in Caco 2 cells was found after addition of Tat. CONCLUSIONS: These results indicate that Tat may be involved in HIV-1-related intestinal disease through direct interaction with enterocytes. PMID- 12557144 TI - Inhaled carbon monoxide suppresses the development of postoperative ileus in the murine small intestine. AB - BACKGROUND & AIMS: The induction of heme oxygenase (HO-1), the rate-limiting enzyme in heme metabolism, is protective against injury in acute and chronic inflammation. Inhalation of low levels of carbon monoxide (CO), a byproduct of heme metabolism, has anti-inflammatory effects equal to HO-1 induction. This study examined whether inhaled CO was protective against the development of postoperative ileus. METHODS: Ileus was induced by surgical anesthesia and gentle manipulation of the mouse small intestine. Animals were exposed to CO (250 ppm) in air 1 hour before and continuously for 24 hours after surgery. RESULTS: CO inhalation prevented the manipulation-induced suppression of circular muscle contractility in vitro, and significantly improved gastrointestinal transit in vivo. Proinflammatory messenger RNA (mRNA) expression (interleukin [IL]-6, IL 1beta, cyclooxygenase 2 [COX-2], inducible nitric oxide [iNOS]) and anti inflammatory mediator expression (IL-10 and HO-1) were elevated 3 to 6 hours after surgery relative to controls. CO treatment reduced IL-1beta and iNOS peak expression by 75%, but not IL-6 or COX-2. In manipulated mice treated with CO, HO 1 expression peaked earlier (3 hours after surgery) and at levels 300% higher than in mice not exposed to CO. IL-10 expression at 3 hours also was 300% higher after CO treatment. CONCLUSIONS: These findings suggest that CO attenuates postoperative ileus by inhibiting selective elements within the inflammatory cascade and by enhanced induction of the anti-inflammatory cytokine IL-10. In addition, the early and enhanced induction of HO-1 potentially amplifies the anti inflammatory effects of the HO-1 pathway by protection from free radical stress and by increasing the tissue availability of CO directly at the sites of inflammation. PMID- 12557145 TI - Bacteroides fragilis enterotoxin induces c-Myc expression and cellular proliferation. AB - BACKGROUND & AIMS: Enterotoxigenic Bacteroides fragilis that secrete a zinc dependent metalloprotease toxin termed the B. fragilis toxin (BFT) have been associated with acute diarrheal disease. BFT rapidly cleaves the extracellular domain of E-cadherin, leading to the complete degradation of the E-cadherin protein. E-cadherin is the primary intercellular adhesion protein of the zonula adherens, and its cytoplasmic domain associates with the nuclear signaling protein beta-catenin. The goal of this study was to examine if BFT triggers beta catenin nuclear signaling in intestinal epithelial cells. METHODS: Cell biologic and biochemical techniques were combined to address beta-catenin nuclear signaling stimulated by BFT. RESULTS: Loss of membrane-associated E-cadherin after BFT treatment of human colonic epithelial cells (HT29/C1 clone) triggers beta-catenin nuclear localization within 3 hours. Subsequently, c-myc transcription and translation are induced and persistent cellular proliferation ensues, mediated in part by beta-catenin/T-cell factor-dependent transcriptional activation. Cellular proliferation is stimulated by as little as 5 x 10(-10) mol/L BFT. CONCLUSIONS: To our knowledge, BFT is the first bacterial toxin reported to activate T-cell factor-dependent beta-catenin nuclear signaling in intestinal epithelial cells. These results suggest that genetic evolution of this common colonic commensal has rendered an organism with the potential to contribute to oncogenic transformation in the colon. PMID- 12557146 TI - Gastric electrical stimulation with short pulses reduces vomiting but not dysrhythmias in dogs. AB - BACKGROUND & AIMS: The aim of this study was to investigate the acute effects of 3 different methods of electrical stimulation in the prevention of vasopressin induced emetic response and gastric dysrhythmias. METHODS: Seven female hound dogs chronically implanted with 4 pairs of electrodes on gastric serosa were used in a 5-session study. Saline and vasopressin were infused in sessions 1 and 2, respectively. In the other 3 sessions with vasopressin infusion, 3 different methods of electrical stimulation (short-pulse stimulation, long-pulse stimulation, and electroacupuncture) were applied. Gastric slow waves and vomiting and behaviors suggestive of nausea were recorded in each session. In a separate study, additional experiments were performed in 5 vagotomized dogs to investigate vagally mediated mechanisms. RESULTS: Vasopressin induced gastric dysrhythmias, uncoupling of slow waves, and vomiting and behaviors suggestive of nausea (P < 0.02, analysis of variance). Long-pulse stimulation, but not short pulse stimulation or electroacupuncture, was capable of preventing vasopressin induced gastric dysrhythmias and gastric slow wave uncoupling. Short-pulse stimulation and electroacupuncture, but not long-pulse stimulation, prevented vomiting and significantly reduced the symptom scores, which was not noted in the dogs with truncal vagotomy. CONCLUSIONS: Long-pulse stimulation normalizes vasopressin-induced slow wave abnormalities with no improvement in vomiting and behaviors suggestive of nausea. Short-pulse stimulation and electroacupuncture prevent vomiting and behaviors suggestive of nausea induced by vasopressin but have no effects on slow waves, and their effects are vagally mediated. PMID- 12557147 TI - Ameliorating effect of anti-inducible costimulator monoclonal antibody in a murine model of chronic colitis. AB - BACKGROUND & AIMS: Inducible costimulator (ICOS)/B7RP-1 represents a newly described receptor/ligand pair involved in costimulation of T cells by antigen presenting cells. We investigated the involvement of the ICOS/B7RP-1 interaction in the pathogenesis of colitis and the therapeutic potential of anti-ICOS monoclonal antibody (mAb) in experimental colitis METHODS: We administered anti ICOS or anti-B7RP-1 mAb to mice with experimental colitis induced by transfer of CD4(+)CD45RB(high) T cells from normal mice into SCID mice. The ability of CD4(+)CD45RB(high) cells derived from ICOS-/- mice to induce colitis was assessed. Th2 cytokine production and apoptosis in infiltrating T cells was examined after administration of anti-ICOS mAb. RESULTS: ICOS was strongly induced on CD4(+) T cells, and B7RP-1 was expressed by macrophages in the inflamed mucosa of colitic mice. Anti-ICOS mAb, but not anti-B7RP-1, ameliorated chronic colitis when administered in prevention or therapeutic protocols. Transfer of CD4(+)CD45RB(high) T cells from ICOS-/- mice induced colitis. Treatment with anti-ICOS mAb did not enhance the production of Th2 cytokines, but a single dose of anti-ICOS mAb induced massive apoptosis of infiltrating ICOS expressing T cells. CONCLUSIONS: ICOS/B7RP-1 interactions are not required for the development of colitis. However, treatment with anti-ICOS mAb can prevent and reverse intestinal inflammation by inducing apoptosis of ICOS-expressing T lymphocytes. PMID- 12557148 TI - Treatment of cirrhosis and liver failure in rats by hepatocyte xenotransplantation. AB - BACKGROUND & AIMS: Hepatocyte transplantation has been proposed as an alternative to liver transplantation for the treatment of hepatic failure. A major limitation to this form of therapy is the availability of human livers as a source of hepatocytes. The use of porcine hepatocytes might address this problem; however, xenogeneic hepatocytes are thought to be functionally incompatible across species and susceptible to irreversible rejection. METHODS: Liver cirrhosis was induced with phenobarbital and carbon tetrachloride. Only rats with decompensated liver failure that did not correct 4 weeks after the discontinuation of carbon tetrachloride were subjected to intrasplenic rat or porcine hepatocyte transplantation. The immunologic integrity of cirrhotic rats was assessed by allogeneic skin grafting, and the immune response to transplanted porcine hepatocytes was assessed by enzyme-linked immunosorbent assay. RESULTS: Porcine hepatocytes restored metabolic function and prolonged the survival of cirrhotic rats, as well as rat hepatocytes. Cirrhotic rats retained the ability to reject allogeneic skin grafts and showed an immune response to the engrafted hepatocytes. Despite this, survival of transplanted porcine hepatocytes was accepted in cirrhotic rats for a period of weeks without immunosuppression. Conventional immunosuppression with FK506 allowed successful retransplantation with hepatocytes from a second porcine donor. CONCLUSIONS: Hepatocytes transplanted between widely divergent species can function to correct liver failure in cirrhotic rats and prolong their survival. Conventional immunosuppression allows long-term functioning of xenogeneic hepatocyte retransplants and suggests that hepatocyte xenotransplantation might be useful as a bridge to liver transplantation and could potentially provide long-term hepatic support. PMID- 12557149 TI - Telomerase reconstitution immortalizes human fetal hepatocytes without disrupting their differentiation potential. AB - BACKGROUND & AIMS: The availability of in vitro expandable human hepatocytes would greatly advance liver-directed cell therapies. Therefore, we examined whether human fetal hepatocytes are amenable to telomerase-mediated immortalization without inducing a transformed phenotype and disrupting their differentiation potential. Telomerase is a ribonucleoprotein that plays a pivotal role in maintaining telomere length and chromosome stability. Human somatic cells, including hepatocytes, exhibit no telomerase activity. Consequently, their telomeres progressively shorten with each cell cycle until critically short telomeres trigger replicative senescence. METHODS: The catalytic subunit, telomerase reverse transcriptase, was expressed in human fetal hepatocytes. Transduced cells were characterized for telomerase activity, telomere length, proliferative capacity, hepatocellular functions, oncogenicity, and their in vivo maturation potential. RESULTS: The expression of human telomerase reverse transcriptase restored telomerase activity in human fetal hepatocytes. Telomerase reconstituted cells were capable of preserving elongated telomeres, propagated in culture beyond replicative senescence for more than 300 cell doublings (to date), and maintained their liver-specific nature, as analyzed by a panel of hepatic growth factors, growth factor receptors, and transcription factors as well as albumin, glucose-6-phosphatase, glycogen synthesis, cytochrome P450 (CYP) expression profiles, and urea production. Moreover, the immortalized cells exhibited no oncogenicity, and no up-regulation of c-Myc was detected. The cells engrafted and survived in the liver of immunodeficient mice with hepatocellular gene expression. CONCLUSIONS: Reconstitution of telomerase activity induces indefinite replication in human fetal hepatocytes and offers unique opportunities for examining basic biologic mechanisms and for considering development of stable cell lines for liver-directed therapies. PMID- 12557150 TI - Delivery of matrix metalloproteinase-1 attenuates established liver fibrosis in the rat. AB - BACKGROUND & AIMS: During hepatic fibrogenesis, the hepatic extracellular matrix changes to fibrillar collagens types I and III, and cirrhosis is believed to produce an irreversible scar. In this study, we investigated whether gene delivery of human matrix metalloproteinase-1, which degrades collagens types I and type III, would attenuate established hepatic fibrosis in the rat, induced by either thioacetamide or bile duct ligation. METHODS: Hepatic fibrosis induced by thioacetamide for 7 weeks was persistent for at least 2 months, even after discontinuation of the treatment. The rats were infected once with a recombinant adenovirus, Ad5MMP-1, into which human pro-human matrix metalloproteinase-1 complementary DNA was packaged, or with a control adenovirus, Ad5LacZ. RESULTS: In Ad5MMP-1-infected, but not in Ad5LacZ-infected, rats, the fibrosis was dramatically attenuated at 2 weeks after the infection. It is interesting to note that the number of activated hepatic stellate cells was also decreased in Ad5MMP 1-infected rats. Moreover, disorganization of the hepatic trabecula, heterogeneity in the size of hepatocytes, and increased dried liver weight were observed only in Ad5MMP-1-treated rats, suggesting that human matrix metalloproteinase-1 stimulated hepatocyte proliferation, which was confirmed by bromodeoxyuridine staining. After 4 weeks, the proliferative effect of human matrix metalloproteinase-1 almost disappeared, but the hepatic fibrosis remained attenuated, whereas the fibrosis in Ad5LacZ-treated rats persisted. Furthermore, the administration of Ad5MMP-1, but not Ad5LacZ, decreased type I collagen and generated a small collagen fragment in hepatic fibrosis induced by bile duct ligation. CONCLUSIONS: Our findings show that transient human matrix metalloproteinase-1 overexpression in the liver effectively attenuates established fibrosis and induces hepatocyte proliferation. PMID- 12557152 TI - Circulating bile salt-dependent lipase originates from the pancreas via intestinal transcytosis. AB - BACKGROUND & AIMS: Bile salt-dependent lipase (BSDL) has been detected in human blood, where it is assumed to play a substantial role in atherosclerosis. The origin of this circulating enzyme is unknown. The aim of this study was to show that blood BSDL originates from pancreatic exocrine secretions via a transcytotic motion across the intestinal epithelium. METHODS: Fluorescein isothiocyanate- or [(125)I]-labeled human pancreatic BSDL was instilled into the lumen of intestinal loops of the rat, and combined biochemical and immunocytochemical investigations were performed in intestinal cells and in the blood of these animals. RESULTS: In vivo pancreatic BSDL is internalized by duodenal enterocytes. The pancreatic enzyme was associated with microvilli and present in endocytic vesicles and Golgi apparatus as well as along the basolateral membrane of enterocytes. It was also detected in intestinal interstitial spaces. Radiolabeled pancreatic BSDL internalized by the duodenal epithelium is the one further detected in circulation. The radiolabeled protein was immunoprecipitated from plasma and had a 100-kilodalton molecular mass compatible with native pancreatic BSDL. In blood, BSDL was mainly associated with low-density lipoproteins. CONCLUSIONS: These in vivo data show that BSDL, normally present in blood, originates from exocrine pancreatic secretion and support the pathophysiologic relevance of BSDL transcytosis through the intestinal mucosa cell lining. Based on this, the implication of circulating BSDL in atherosclerosis merits careful attention. PMID- 12557151 TI - Antiproliferative property of sphingosine 1-phosphate in rat hepatocytes involves activation of Rho via Edg-5. AB - BACKGROUND & AIMS: Sphingosine 1-phosphate (S1P), a ligand for G protein-coupled endothelial differentiation gene-1 (Edg-1), Edg-3, Edg-5, Edg-6, and Edg-8, elicits a variety of responses by cells. Prominent among these is cell proliferation. S1P is abundantly stored in platelets and released upon their activation, suggesting that S1P plays a pathophysiologic role in vivo. Because the major part of injected S1P was distributed into the liver in mice, we wondered whether the liver would be one of its targets. The effects of S1P on hepatocytes, the major constituent cells in the liver, were examined. METHODS & RESULTS: Northern blot analysis revealed the expression of Edg-1 and Edg-5 messenger RNA (mRNA) in cultured rat hepatocytes, in which S1P decreased DNA synthesis induced by hepatocyte growth factor (HGF) or epidermal growth factor (EGF) without affecting total protein synthesis. This inhibitory effect was attenuated by inactivation of small GTPase Rho with C3 exotoxin but not by inactivation of G(i) with pertussis toxin. Moreover, in the presence of JTE-013, a newly developed and specific binding antagonist for Edg-5, the inhibitory effect was also cancelled. Finally, the administration of S1P after 70% partial hepatectomy in rats reduced the peak of DNA synthesis in hepatocytes with increased Rho activity. Furthermore, Edg-5 but not Edg-1 mRNA expression was enhanced in hepatocytes 24-72 hours after partial hepatectomy, which coincides with decreasing hepatocyte proliferation. CONCLUSIONS: S1P has an antiproliferative property in rat hepatocytes by activating Rho via Edg-5. Our results raise the possibility that S1P is a negative regulator in liver regeneration. PMID- 12557153 TI - Severe acute pancreatitis and reduced acinar cell apoptosis in the exocrine pancreas of mice deficient for the Cx32 gene. AB - BACKGROUND & AIMS: The early events leading to acinar cell injury during acute pancreatitis are poorly characterized. Signaling through gap junction channels contributes to the homeostasis of the exocrine pancreas by coordinating acinar cell activity within an acinus. To explore the role of gap junctional communication in acinar cell response to injury, we analyzed the course of acute pancreatitis induced by injection of cerulein in mice deficient for Cx32, the major gap junction protein expressed in the exocrine pancreas. METHODS: The severity of pancreatitis was evidenced by measuring serum amylase activity, pancreatic edema, acinar cell necrosis, pancreatic tumor necrosis factor alpha concentration, and myeloperoxidase activity. Acinar cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL), caspase-3 activity, and Bax/Bcl-2 expression. Expression and function of connexin were evaluated by immunofluorescence and dye coupling. RESULTS: Cx32-deficient mice exhibited a deleterious course of acute pancreatitis with increased necrosis, edema, and inflammation of the exocrine pancreas. In addition, the exocrine pancreas of Cx32-deficient mice showed a decreased number of TUNEL-positive acinar cells and decreased caspase-3 activity but no change in Bax or Bcl-2 pancreatic expression. Interestingly, chemicals known to induce apoptosis in vivo had no effect on Cx32-deficient pancreatic acinar cells. CONCLUSIONS: Deficiency of a pancreatic connexin converts a mild reversible form of acute pancreatitis into a severe disease and decreases the sensitivity of acinar cells to apoptotic stimuli. The results show that acinar cell-to-cell communication plays a key role in the modulation of severity of acute pancreatitis. PMID- 12557154 TI - Mitochondrial permeability transition in the switch from necrotic to apoptotic cell death in ischemic rat hepatocytes. AB - BACKGROUND & AIMS: Ischemia/reperfusion can initiate both necrotic and apoptotic death of hepatocytes. Previous work showed that onset of the mitochondrial permeability transition (MPT) can initiate necrotic cell death after reperfusion, but the MPT is also implicated in apoptosis. Here, we investigated factors regulating how cell death switches from necrosis to apoptosis after ischemia/reperfusion injury. METHODS: Overnight cultured rat hepatocytes were incubated in anoxia at pH 6.2 for 4 hours and reoxygenated at pH 7.4 to simulate ischemia/reperfusion. Some cells were incubated with fructose plus glycine just before and then continuously after reperfusion. Development of apoptosis was evaluated by examining chromatin condensation, nuclear DNA fragmentation, and caspase 3 activity. RESULTS: Reperfusion with the glycolytic substrate fructose plus the cytoprotective amino acid glycine prevented necrotic cell killing. Instead, apoptosis developed within 12 hours as shown by nuclear chromatin changes, TUNEL staining, and caspase 3 activation. This apoptotic cell killing was prevented by cyclosporin A, an MPT blocker, and by pancaspase and caspase 3 inhibition, but not by caspase 8 inhibition. Cyclosporin A also blocked caspase-3 activation. Reperfusion with glycine alone prevented necrotic cell death but did not induce apoptosis and only poorly promoted recovery of ATP, whereas fructose alone during reperfusion promoted both ATP recovery and apoptosis. CONCLUSIONS: Glycolytic ATP generation after reperfusion prevents necrotic killing of hepatocytes after simulated ischemia/reperfusion despite onset of the MPT. Instead, the MPT promotes caspase-and ATP-dependent apoptosis. Thus, the MPT is a common mechanism responsible for both necrosis and apoptosis after ischemia/reperfusion. PMID- 12557156 TI - The genetics of inflammatory bowel disease. AB - The inflammatory bowel diseases (IBD) comprise complex genetic disorders, with multiple contributing genes. Linkage studies have implicated several genomic regions as likely containing IBD susceptibility genes, with some observed uniquely in Crohn's disease (CD) or ulcerative colitis (UC), and others common to both disorders. The best replicated linkage region, IBD1, on chromosome 16q contains the CD susceptibility gene, NOD2/CARD15. NOD2/CARD15 is expressed in peripheral blood monocytes and is structurally related to the plant R proteins, which mediate host resistance to microbial pathogens. Three major coding region polymorphisms within NOD2/CARD15 have been highly associated with CD among patients of European descent. Having one copy of the risk alleles confers a 2-4 fold risk for developing CD, whereas double-dose carriage increases the risk 20 40-fold. All 3 major CD variants exhibit a deficit in NF-kappaB activation in response to bacterial components. Carriage of NOD2/CARD15 risk alleles is associated with ileal location, earlier disease onset, and stricturing phenotype. Other IBD genomic regions include IBD2 on chromosome 12q (observed more in UC), and IBD3, containing the major histocompatibility complex region. A short genomic region has been associated with CD on chromosome 5q, but the precise contributing gene is as yet unidentified. The characterization of additional IBD susceptibility genes could potentially lead to the identification of novel therapeutic agents for IBD, make possible a molecular reclassification of disease, and increase understanding of the contribution of environmental factors (notably, tobacco and the intestinal microbial milieu) to intestinal inflammation. PMID- 12557155 TI - Antifibrogenic effects of canrenone, an antialdosteronic drug, on human hepatic stellate cells. AB - BACKGROUND & AIMS: Several lines of evidence indicate that aldosterone antagonists may exert direct antifibrogenic effects. The aim of this study was to evaluate the possible direct antifibrogenic effects of canrenone, the active metabolite of spironolactone, in activated human hepatic stellate cells. METHODS: The effects of canrenone were assessed on platelet-derived growth factor-induced mitogenic and chemotactic effects and the increased de novo synthesis of different extracellular matrix components induced by transforming growth factor beta1. RESULTS: Canrenone dose-dependently reduced platelet-derived growth factor induced cell proliferation and motility. This effect was not associated with either changes in the phosphorylation of platelet-derived growth factor receptor and phospholipase C gamma or in the activation of the Ras/extracellular signal regulated kinase pathway, whereas it was accompanied by a dose-dependent inhibition of platelet-derived growth factor-induced phosphatidylinositol 3 kinase activity. In addition, canrenone inhibited the activity of the Na(+)/H(+) exchanger 1 induced by platelet-derived growth factor. The effect of canrenone on Na(+)/H(+) exchanger 1 activity was reproduced by phosphatidylinositol 3-kinase inhibitors, thus supporting an inhibitory action of canrenone on phosphatidylinositol 3-kinase activity. To further address this possibility, the action of canrenone was compared with that of 2 established Na(+)/H(+) exchanger 1 inhibitors: ethylisopropylamiloride and cariporide. Whereas ethylisopropylamiloride was able to inhibit platelet-derived growth factor induced phosphatidylinositol 3-kinase activity, cariporide was without any effect. Both compounds reproduced the effects of canrenone on platelet-derived growth factor-induced mitogenesis and chemotaxis. Finally, canrenone was able to reduce transforming growth factor-beta1-induced de novo synthesis of procollagen type I/IV and fibronectin and thrombin-induced hepatic stellate cell contraction. CONCLUSIONS: These results indicate that canrenone may be active as an antifibrogenic drug. PMID- 12557157 TI - Consecutive regression of concurrent laryngeal and gastric MALT lymphoma after anti-Helicobacter pylori therapy. AB - The most common primary lymphoma of the gastrointestinal tract is B-cell lymphoma arising from mucosa-associated lymphoid tissue known as MALT lymphoma. Although the majority of these lesions affect the stomach and are associated with Helicobacter pylori organisms, sites other than the gastrointestinal tract may be affected. This case report describes a patient with concomitant laryngeal MALT lymphoma and Helicobacter pylori-related gastric MALT lymphoma derived from the same clone as confirmed by PCR. Treatment of Helicobacter pylori infection in this patient using antibiotics led to regression of both lesions. This patient remains in remission at 46-month follow-up. This is the first case report on the regression of a laryngeal MALT lymphoma after Helicobacter pylori eradication. We suggest that all patients presenting with extragastric MALT lymphoma should undergo upper gastrointestinal endoscopy with gastric biopsies for the determination of Helicobacter pylori status and presence of concomitant gastric MALT lymphoma, followed by a course of anti-Helicobacter pylori antibiotic therapy. Nonresponders may subsequently be considered for surgery and/or chemo/radiation therapy. PMID- 12557158 TI - Colorectal cancer screening and surveillance: clinical guidelines and rationale Update based on new evidence. AB - We have updated guidelines for screening for colorectal cancer. The original guidelines were prepared by a panel convened by the U.S. Agency for Health Care Policy and Research and published in 1997 under the sponsorship of a consortium of gastroenterology societies. Since then, much has changed, both in the research rature and in the clinical context. The present report summarizes new developments in this field and suggests how they should change practice. As with the previous version, these guidelines offer screening options and encourage the physician and patient to decide together which is the best approach for them. The guidelines also take into account not only the effectiveness of screening but also the risks, inconvenience, and cost of the various approaches. These guidelines differ from those published in 1997 in several ways: we recommend against rehydrating fecal occult blood tests; the screening interval for double contrast barium enema has been shortened to 5 years; colonoscopy is the preferred test for the diagnostic investigation of patients with findings on screening and for screening patients with a family history of hereditary nonpolyposis colorectal cancer; recommendations for people with a family history of colorectal cancer make greater use of risk stratification; and guidelines for genetic testing are included. Guidelines for surveillance are also included. Follow-up of postpolypectomy patients relies now on colonoscopy, and the first follow-up examination has been lengthened from 3 to 5 years for low-risk patients. If this were adopted nationally, surveillance resources could be shifted to screening and diagnosis. Promising new screening tests (virtual colonoscopy and tests for altered DNA in stool) are in development but are not yet ready for use outside of research studies. Despite a consensus among expert groups on the effectiveness of screening for colorectal cancer, screening rates remain low. Improvement depends on changes in patients' attitudes, physicians' behaviors, insurance coverage, and the surveillance and reminder systems necessary to support screening programs. PMID- 12557159 TI - Tales from the crypt. PMID- 12557160 TI - PPARgamma ligands: taking Ppart in chemoprevention. PMID- 12557161 TI - Hepatocyte transplantation: new possibilities for therapy. PMID- 12557162 TI - An aspirin a day keeps the cardiologist away (but what about the gastroenterologist?). PMID- 12557164 TI - The mesalamine wars heat up-enter balsalazide. PMID- 12557165 TI - The stomach and weight reduction: the role of ghrelin. PMID- 12557166 TI - The politics of H. pylori-challenged mice: liberal permissiveness versus conservative restriction. PMID- 12557171 TI - Discussion on the true impact of fatigue in primary biliary cirrhosis: a population study. PMID- 12557172 TI - Anti-CD2O chimeric monoclonal antibody (rituximab) treatment of immune-mediated thrombocytopenia associated with Crohn's disease. PMID- 12557173 TI - Serine protease inhibitors cause F-actin redistribution in pancreatic acini? PMID- 12557174 TI - PEG-interferon versus conventional interferon and liver fibrosis: do we have evidence of superiority? PMID- 12557176 TI - Serologic tests for celiac disease. PMID- 12557178 TI - Tenofovir disoproxil fumarate exhibits strong antiviral effect in a patient with lamivudine-resistant severe hepatitis B reactivation. PMID- 12557181 TI - Basic interdomain boundary residues in calmodulin decrease calcium affinity of sites I and II by stabilizing helix-helix interactions. AB - Calmodulin is an EF-hand calcium-binding protein (148 a.a.) essential in intracellular signal transduction. Its homologous N- and C-terminal domains are separated by a linker that appears disordered in NMR studies. In a study of an N domain fragment of Paramecium CaM (PCaM1-75), the addition of linker residues 76 to 80 (MKEQD) raised the Tm by 9 degrees C and lowered calcium binding by 0.54 kcal/mol (Sorensen et al., [Biochemistry 2002;41:15-20]), showing that these tether residues affect energetics as well as being a barrier to diffusion. To determine the individual contributions of residues 74 through 80 (RKMKEQD) to stability and calcium affinity, we compared a nested series of 7 fragments (PCaM1 74 to PCaM1-80). For the first 4, PCaM1-74 through PCaM1-77, single amino acid additions at the C-terminus corresponded to stepwise increases in thermostability and decreases in calcium affinity with a net change of 13.5 degrees C in Tm and 0.55 kcal/mol in free energy. The thermodynamic properties of fragments PCaM1-77 through PCaM1-80 were nearly identical. We concluded that the 3 basic residues in the sequence from 74 to 77 (RKMK) are critical to the increased stability and decreased calcium affinity of the longer N-domain fragments. Comparisons of NMR (HSQC) spectra of 15N-PCaM1-74 and 15N-PCaM1-80 and analysis of high-resolution structural models suggest these residues are latched to amino acids in helix A of CaM. The addition of residues E78, Q79, and D80 had a minimal effect on sites I and II, but they may contribute to the mechanism of energetic communication between the domains. PMID- 12557182 TI - Strategies for structural proteomics of prokaryotes: Quantifying the advantages of studying orthologous proteins and of using both NMR and X-ray crystallography approaches. AB - Only about half of non-membrane-bound proteins encoded by either bacterial or archaeal genomes are soluble when expressed in Escherichia coli (Yee et al., Proc Natl Acad Sci USA 2002;99:1825-1830; Christendat et al., Prog Biophys Mol Biol 200;73:339-345). This property limits genome-scale functional and structural proteomics studies, which depend on having a recombinant, soluble version of each protein. An emerging strategy to increase the probability of deriving a soluble derivative of a protein is to study different sequence homologues of the same protein, including representatives from thermophilic organisms, based on the assumption that the stability of these proteins will facilitate structural analysis. To estimate the relative merits of this strategy, we compared the recombinant expression, solubility, and suitability for structural analysis by NMR and/or X-ray crystallography for 68 pairs of homologous proteins from E. coli and Thermotoga maritima. A sample suitable for structural studies was obtained for 62 of the 68 pairs of homologs under standardized growth and purification procedures. Fourteen (eight E. coli and six T. maritima proteins) samples generated NMR spectra of a quality suitable for structure determination and 30 (14 E. coli and 16 T. maritima proteins) samples formed crystals. Only three (one E. coli and two T. maritima proteins) samples both crystallized and had excellent NMR properties. The conclusions from this work are: (1) The inclusion of even a single ortholog of a target protein increases the number of samples for structural studies almost twofold; (2) there was no clear advantage to the use of thermophilic proteins to generate samples for structural studies; and (3) for the small proteins analyzed here, the use of both NMR and crystallography approaches almost doubled the number of samples for structural studies. PMID- 12557183 TI - Alpha helix shortening in 1522 MSP-1 conserved peptide analogs is associated with immunogenicity and protection against P. falciparum malaria. AB - 1522 is a nonimmunogenic conserved high-activity binding peptide (HABP) belonging to Plasmodium falciparum MSP-1 protein N-terminal fragment. The key amino acids in binding to red blood cells (RBC) were identified and replaced by others having similar mass but different charge. Because conserved HABPs are not antigenic nor immunogenic, immunogenicity and protectivity studies were then conducted on them in the Aotus monkey. 1H-NMR studies included the lead peptide 1522 as well as the analogs 9782, 13446, 13448, and 13442 to relate their structure to biological function. All the peptides presented alpha-helical structure, with differences observed in helix location and extension. The nonprotective 1522 peptide was totally helical from the N- to the C-terminus, very similar to nonprotective 13442 and 13448 peptides whose extension was almost totally helical. The 9782 and 13446 protective peptides, however, possessed a shorter helical region where modified critical binding residues were not included. A more flexible region was generated at the C-terminus in those peptides with a shorter helical region, leading to a greater number of conformers. These data suggest that peptide flexibility results in increased interaction with immune system molecules, generating protective immunity. PMID- 12557184 TI - Structure and dynamics of the potato carboxypeptidase inhibitor by 1H and 15N NMR. AB - The solution structure and backbone dynamics of the recombinant potato carboxypeptidase inhibitor (PCI) have been characterized by NMR spectroscopy. The structure, determined on the basis of 497 NOE-derived distance constraints, is much better defined than the one reported in a previous NMR study, with an average pairwise backbone root-mean-square deviation of 0.5 A for the well defined region of the protein, residues 7-37. Many of the side-chains show now well-defined conformations, both in the hydrophobic core and on the surface of the protein. Overall, the solution structure of free PCI is similar to the one that it shows in the crystal of the complex with carboxypeptidase A. However, some local differences are observed in regions 15-21 and 27-29. In solution, the six N-terminal and the two C-terminal residues are rather flexible, as shown by 15N backbone relaxation measurements. The flexibility of the latter segment may have implications in the binding of the inhibitor by the enzyme. All the remaining residues in the protein are essentially rigid (S2 > 0.8) with the exception of two of them at the end of a short 3/10 helix. Despite the small size of the protein, a number of amide protons are protected from exchange with solvent deuterons. The slowest exchanging protons are those in a small two-strand beta-sheet. The unfolding free energies, as calculated from the exchange rates of these protons, are around 5 kcal/mol. Other protected amide protons are located in the segment 7-12, adjacent to the beta-sheet. Although these residues are not in an extended conformation in PCI, the equivalent residues in structurally homologous proteins form a third strand of the central beta-sheet. The amide protons in the 3/10 helix are only marginally protected, indicating that they exchange by a local unfolding mechanism, which is consistent with the increase in flexibility shown by some of its residues. Backbone alignment-based programs for folding recognition, as opposite to disulfide-bond alignments, reveal new proteins of unrelated sequence and function with a similar structure. PMID- 12557185 TI - Structure of the Clade 1 catalase, CatF of Pseudomonas syringae, at 1.8 A resolution. AB - Catalase CatF of Pseudomonas syringae has been identified phylogenetically as a clade 1 catalase, closely related to plant catalases, a group from which no structure has been determined. The structure of CatF has been refined at 1.8 A resolution by using X-ray synchrotron data collected from a crystal flash-cooled with liquid nitrogen. The crystallographic agreement factors R and R(free) are, respectively, 18.3% and 24.0%. The asymmetric unit of the crystal contains a whole molecule that shows accurate 222-point group symmetry. The crystallized enzyme is a homotetramer of subunits with 484 residues, some 26 residues shorter than predicted from the DNA sequence. Mass spectrometry analysis confirmed the absence of 26 N-terminal residues, possibly removed by a periplasmic transport system. The core structure of the CatF subunit was closely related to seven other catalases with root-mean-square deviations (RMSDs) of 368 core Calpha atoms of 0.99-1.30 A. The heme component of CatF is heme b in the same orientation that is found in Escherichia coli hydroperoxidase II, an orientation that is flipped 180 degrees with respect the orientation of the heme in bovine liver catalase. NADPH is not found in the structure of CatF because key residues required for nucleotide binding are missing; 2129 water molecules were refined into the model. Water occupancy in the main or perpendicular channel of CatF varied among the four subunits from two to five in the region between the heme and the conserved Asp150. A comparison of the water occupancy in this region with the same region in other catalases reveals significant differences among the catalases. PMID- 12557187 TI - Comparison of a QM/MM force field and molecular mechanics force fields in simulations of alanine and glycine "dipeptides" (Ace-Ala-Nme and Ace-Gly-Nme) in water in relation to the problem of modeling the unfolded peptide backbone in solution. AB - We compare the conformational distributions of Ace-Ala-Nme and Ace-Gly-Nme sampled in long simulations with several molecular mechanics (MM) force fields and with a fast combined quantum mechanics/molecular mechanics (QM/MM) force field, in which the solute's intramolecular energy and forces are calculated with the self-consistent charge density functional tight binding method (SCCDFTB), and the solvent is represented by either one of the well-known SPC and TIP3P models. All MM force fields give two main states for Ace-Ala-Nme, beta and alpha separated by free energy barriers, but the ratio in which these are sampled varies by a factor of 30, from a high in favor of beta of 6 to a low of 1/5. The frequency of transitions between states is particularly low with the amber and charmm force fields, for which the distributions are noticeably narrower, and the energy barriers between states higher. The lower of the two barriers lies between alpha and beta at values of psi near 0 for all MM simulations except for charmm22. The results of the QM/MM simulations vary less with the choice of MM force field; the ratio beta/alpha varies between 1.5 and 2.2, the easy pass lies at psi near 0, and transitions between states are more frequent than for amber and charmm, but less frequent than for cedar. For Ace-Gly-Nme, all force fields locate a diffuse stable region around phi = pi and psi = pi, whereas the amber force field gives two additional densely sampled states near phi = +/-100 degrees and psi = 0, which are also found with the QM/MM force field. For both solutes, the distribution from the QM/MM simulation shows greater similarity with the distribution in high-resolution protein structures than is the case for any of the MM simulations. PMID- 12557186 TI - Structure validation by Calpha geometry: phi,psi and Cbeta deviation. AB - Geometrical validation around the Calpha is described, with a new Cbeta measure and updated Ramachandran plot. Deviation of the observed Cbeta atom from ideal position provides a single measure encapsulating the major structure-validation information contained in bond angle distortions. Cbeta deviation is sensitive to incompatibilities between sidechain and backbone caused by misfit conformations or inappropriate refinement restraints. A new phi,psi plot using density dependent smoothing for 81,234 non-Gly, non-Pro, and non-prePro residues with B < 30 from 500 high-resolution proteins shows sharp boundaries at critical edges and clear delineation between large empty areas and regions that are allowed but disfavored. One such region is the gamma-turn conformation near +75 degrees,-60 degrees, counted as forbidden by common structure-validation programs; however, it occurs in well-ordered parts of good structures, it is overrepresented near functional sites, and strain is partly compensated by the gamma-turn H-bond. Favored and allowed phi,psi regions are also defined for Pro, pre-Pro, and Gly (important because Gly phi,psi angles are more permissive but less accurately determined). Details of these accurate empirical distributions are poorly predicted by previous theoretical calculations, including a region left of alpha helix, which rates as favorable in energy yet rarely occurs. A proposed factor explaining this discrepancy is that crowding of the two-peptide NHs permits donating only a single H-bond. New calculations by Hu et al. [Proteins 2002 (this issue)] for Ala and Gly dipeptides, using mixed quantum mechanics and molecular mechanics, fit our nonrepetitive data in excellent detail. To run our geometrical evaluations on a user-uploaded file, see MOLPROBITY (http://kinemage.biochem.duke.edu) or RAMPAGE (http://www cryst.bioc.cam.ac.uk/rampage). PMID- 12557188 TI - Comparative properties of a three-dimensional model of Plasmodium falciparum ornithine decarboxylase. AB - The ornithine decarboxylase (ODC) component of the bifunctional S adenosylmethionine decarboxylase/ornithine decarboxylase enzyme (PfAdoMetDC-ODC) of Plasmodium falciparum was modeled on the crystal structure of the Trypanosoma brucei enzyme. The homology model predicts a doughnut-shaped active homodimer that associates in a head-to-tail manner. The monomers contain two distinct domains, an N-terminal alpha/beta-barrel and a C-terminal modified Greek-key domain. These domains are structurally conserved between eukaryotic ODC enzymes and are preserved in distant analogs such as alanine racemase and triosephosphate isomerase-like proteins. Superimposition of the PfODC model on the crystal structure of the human enzyme indicates a significant degree of deviation in the carbon alpha-backbone of the solvent accessible loops. The surface locality of the ab initio modeled 38 amino acid parasite-specific insert suggests a role in the stabilization of the large bifunctional protein complex. The active site pockets of PfODC at the interface between the monomers appear to be conserved regarding the binding sites of the cofactor and substrate, but each contains five additional malaria-specific residues. The predicted PfODC homology model is consistent with mutagenesis results and biochemical studies concerning the active site residues and areas involved in stabilizing the dimeric form of the protein. Two competitive inhibitors of PfODC could be shown to interact with several parasite-specific residues in comparison with their interaction with the human ODC. The PfODC homology model contributes toward a structure-based approach for the design of novel malaria-specific inhibitors. PMID- 12557189 TI - Structural studies on MtRecA-nucleotide complexes: insights into DNA and nucleotide binding and the structural signature of NTP recognition. AB - RecA protein plays a crucial role in homologous recombination and repair of DNA. Central to all activities of RecA is its binding to Mg(+2)-ATP. The active form of the protein is a helical nucleoprotein filament containing the nucleotide cofactor and single-stranded DNA. The stability and structure of the helical nucleoprotein filament formed by RecA are modulated by nucleotide cofactors. Here we report crystal structures of a MtRecA-ADP complex, complexes with ATPgammaS in the presence and absence of magnesium as well as a complex with dATP and Mg+2. Comparison with the recently solved crystal structures of the apo form as well as a complex with ADP-AlF4 confirms an expansion of the P-loop region in MtRecA, compared to its homologue in Escherichia coli, correlating with the reduced affinity of MtRecA for ATP. The ligand bound structures reveal subtle variations in nucleotide conformations among different nucleotides that serve in maintaining the network of interactions crucial for nucleotide binding. The nucleotide binding site itself, however, remains relatively unchanged. The analysis also reveals that ATPgammaS rather than ADP-AlF4 is structurally a better mimic of ATP. From among the complexed structures, a definition for the two DNA-binding loops L1 and L2 has clearly emerged for the first time and provides a basis to understand DNA binding by RecA. The structural information obtained from these complexes correlates well with the extensive biochemical data on mutants available in the literature, contributing to an understanding of the role of individual residues in the nucleotide binding pocket, at the molecular level. Modeling studies on the mutants again point to the relative rigidity of the nucleotide binding site. Comparison with other NTP binding proteins reveals many commonalties in modes of binding by diverse members in the structural family, contributing to our understanding of the structural signature of NTP recognition. PMID- 12557190 TI - Repair of impurity-poisoned protein crystal surfaces. AB - The surface morphology of Bence-Jones protein (BJP) crystals was investigated during growth and dissolution by using in situ atomic force microscopy (AFM). It was shown that over a wide supersaturation range, impurities adsorb on the crystalline surface and ultimately form an impurity adsorption layer that prevents further growth of the crystal. At low undersaturations, this impurity adsorption layer prevents dissolution. At greater undersaturation, dissolution takes place around large particles incorporated into the crystal, leading to etch pits with impurity-free bottoms. On restoration of supersaturation conditions, two-dimensional nucleation takes place on the impurity-free bottoms of these etch pits. After new growth layers fill in the etch pits, they cover the impurity poisoned top layer of the crystal face. This leads to the resumption of its growth. Formation of an impurity-adsorption layer can explain the termination of growth of macromolecular crystals that has been widely noted. Growth-dissolution growth cycles could be used to produce larger crystals that otherwise would have stopped growing because of impurity poisoning. PMID- 12557191 TI - Refinement of protein structures in explicit solvent. AB - We present a CPU efficient protocol for refinement of protein structures in a thin layer of explicit solvent and energy parameters with completely revised dihedral angle terms. Our approach is suitable for protein structures determined by theoretical (e.g., homology modeling or threading) or experimental methods (e.g., NMR). In contrast to other recently proposed refinement protocols, we put a strong emphasis on consistency with widely accepted covalent parameters and computational efficiency. We illustrate the method for NMR structure calculations of three proteins: interleukin-4, ubiquitin, and crambin. We show a comparison of their structure ensembles before and after refinement in water with and without a force field energy term for the dihedral angles; crambin was also refined in DMSO. Our results demonstrate the significant improvement of structure quality by a short refinement in a thin layer of solvent. Further, they show that a dihedral angle energy term in the force field is beneficial for structure calculation and refinement. We discuss the optimal weight for the energy constant for the backbone angle omega and include an extensive discussion of meaning and relevance of the calculated validation criteria, in particular root mean square Z scores for covalent parameters such as bond lengths. PMID- 12557192 TI - Molecular dynamics simulation reveals a surface salt bridge forming a kinetic trap in unfolding of truncated Staphylococcal nuclease. AB - Surface salt bridges are ubiquitous in globular proteins. Their contribution to protein stability has been extensively debated in the past decade. Here, molecular dynamics simulations are performed starting from a non-equilibrium state of Staphylococcal nuclease (SNase) with C-terminal truncation (SNaseDelta). The results indicate a key role in the unfolding of the surface salt bridge between arginine 105 and glutamate 135. Experimentally, SNaseDelta is known to be partially unfolded. However, in simulations over 1 ns at 300 K and over 500 ps at 400 K, SNaseDelta remains stable in the native-like folded conformation, the salt bridge hindering unfolding. When the potential function is altered so as to selectively weaken the salt bridge, which then breaks rapidly at 430 K, the protein starts to unfold. The results suggest that breaking of this salt bridge presents a significant barrier to the unfolding transition of SNaseDelta from a native-like state to the unfolded state. Potential of mean force calculations indicate that the barrier height for this transition is approximately 7 kcal/mol. PMID- 12557193 TI - Hydration in proteins observed by high-resolution neutron crystallography. AB - It is well known that water molecules surrounding a protein play important roles in maintaining its structural stability. Water molecules are known to participate in several physiological processes through the formation of hydrogen bonds. However, the hydration structures of most proteins are not known well at an atomic level at present because X-ray protein crystallography has difficulties to localize hydrogen atoms. In contrast, neutron crystallography has no problem in determining the position of hydrogens with high accuracy.1 In this article, the hydration structures of three proteins are described- myoglobin, wild-type rubredoxin, and a mutant rubredoxin-the structures of which were solved at 1.5- or 1.6-A resolution by neutron structure determination. These hydration patterns show fascinating features and the water molecules adopt a variety of shapes in the neutron Fourier maps, revealing details of intermolecular hydrogen bond formation and dynamics of hydration. Our results further show that there are strong relationships between these shapes and the water environments. PMID- 12557195 TI - Regeneration research today. PMID- 12557196 TI - Muscle regeneration in amphibians and mammals: passing the torch. AB - Skeletal muscle in both amphibians and mammals possesses a high regenerative capacity. In amphibians, a muscle can regenerate in two distinct ways: as a tissue component of an entire regenerating limb (epimorphic regeneration) or as an isolated entity (tissue regeneration). In the absence of epimorphic regenerative ability, mammals can regenerate muscles only by the tissue mode. This review focuses principally on the regeneration of entire muscles and covers what is known and what remains to be elucidated about fundamental mechanisms underlying muscle regeneration at this level. PMID- 12557197 TI - Hydra regeneration and epitheliopeptides. AB - Hydra has been well known for over 200 years for its remarkable regenerative capacity. In addition to small pieces excised from the body, reaggregates of dissociated single cells can also regenerate. Although the cellular events involved in the regeneration process have been well characterized, the underlying molecular mechanisms are yet to be uncovered. Recently, however, transcription factors and signaling molecules, both proteins and short peptides, have been identified and their role suggested in patterning and morphogenesis. In this article, a regeneration study at the tissue level is first described and then the importance of epithelial cells in regeneration is stressed. Finally, the recent study on morphogenetic peptides derived from epithelial cells is reviewed. PMID- 12557198 TI - Old questions, new tools, and some answers to the mystery of fin regeneration. AB - Pluridisciplinary approaches led to the notion that fin regeneration is an intricate phenomenon involving epithelial-mesenchymal and reciprocal exchanges throughout the process as well as interactions between ray and interray tissue. The establishment of a blastema after fin amputation is the first event leading to the reconstruction of the missing part of the fin. Here, we review our knowledge on the origin of the blastema, its formation and growth, and of the mechanisms that control differentiation and patterning of the regenerate. Our current understanding results from studies of fin regeneration performed in various teleost fish over the past century. We also report the recent breakthroughs that have been made in the past decade with the arrival of a new model, the zebrafish, Danio rerio, which now offers the possibility to combine cytologic, molecular, and genetic analyses and open new perspectives in this field. PMID- 12557199 TI - Tales of regeneration in zebrafish. AB - Complex tissue regeneration involves exquisitely coordinated proliferation and patterning of adult cells after severe injury or amputation. Certain lower vertebrates such as urodele amphibians and teleost fish have a greater capacity for regeneration than mammals. However, little is known about molecular mechanisms of regeneration, and cellular mechanisms are incompletely defined. To address this deficiency, we and others have focused on the zebrafish model system. Several helpful tools and reagents are available for use with zebrafish, including the potential for genetic approaches to regeneration. Recent studies have shed light on the remarkable ability of zebrafish to regenerate fins. PMID- 12557200 TI - Eye regeneration at the molecular age. AB - Eye tissues such as the lens and the retina possess remarkable regenerative abilities. In amphibians, a complete lens can be regenerated after lentectomy. The process is a classic example of transdifferentiation of one cell type to another. Likewise, retina can be regenerated, but the strategy used to replace the damaged retina differs, depending on the animal system and the age of the animal. Retina can be regenerated by transdifferentiation or by the use of stem cells. In this review, we present a synthesis on the regenerative capacity of eye tissues in different animals with emphasis on the strategy and the molecules involved. In addition, we stress the place of this field at the molecular age and the importance of the recent technologic advances. PMID- 12557201 TI - Head regeneration in Hydra. AB - Hydra, a primitive metazoan, has a simple structure consisting of a head, body column, and foot aligned along a single oral-aboral axis. The body column has a high capacity for regeneration of both the head and foot. Because of the tissue dynamics that take place in adult Hydra, the processes governing axial patterning are continuously active to maintain the form of the animal. Regeneration in hydra is morphallactic and closely related to these axial patterning processes. As might be expected, analysis at the molecular level indicates that the same set of genes are involved in head regeneration and the maintenance of the head in the context of the tissue dynamics of the adult. The genes analyzed so far play roles in axial patterning processes in bilaterians. PMID- 12557202 TI - Retinoic acid, a regeneration-inducing molecule. AB - Retinoic acid (RA) is the biologically active metabolite of vitamin A. It is a low molecular weight, lipophilic molecule that acts on the nucleus to induce gene transcription. In amphibians and mammals, it induces the regeneration of several tissues and organs and these examples are reviewed here. RA induces the "super regeneration" of organs that can already regenerate such as the urodele amphibian limb by respecifying positional information in the limb. In organs that cannot normally regenerate such as the adult mammalian lung, RA induces the complete regeneration of alveoli that have been destroyed by various noxious treatments. In the mammalian central nervous system (CNS), which is another tissue that cannot regenerate, RA does not induce neurite outgrowth as it does in the embryonic CNS, because one of the retinoic acid receptors, RAR beta 2, is not up regulated. When RAR beta 2 is transfected into the adult spinal cord in vitro, then neurite outgrowth is stimulated. In all these cases, RA is required for the development of the organ, in the first place suggesting that the same gene pathways are likely to be used for both development and regeneration. This suggestion, therefore, might serve as a strategy for identifying potential tissue or organ targets that have the capacity to be stimulated to regenerate. PMID- 12557203 TI - Changes in spinal cord regenerative ability through phylogenesis and development: lessons to be learnt. AB - Lower vertebrates, such as fish and amphibians, and developing higher vertebrates can regenerate complex body structures, including significant portions of their central nervous system. It is still poorly understood why this potential is lost with evolution and development and becomes very limited in adult mammals. In this review, we will discuss the current knowledge on the cellular and molecular changes after spinal cord injury in adult tailed amphibians, where regeneration does take place, and in developing chick and mammalian embryos at different developmental stages. We will focus on the recruitment of progenitor cells to repair the damage and discuss possible roles of changes in early response to injury, such as cell death by apoptosis, and of myelin-associated proteins, such as Nogo, in the transition between regeneration-competent and regeneration incompetent stages of development. A better understanding of the mechanisms underlying spontaneous regeneration of the spinal cord in vivo in amphibians and in the chick embryo will help to devise strategies for restoring function to damaged or diseased nervous tissues in mammals. PMID- 12557204 TI - Cnidarians: an evolutionarily conserved model system for regeneration? AB - Cnidarians are among the simplest metazoan animals and are well known for their remarkable regeneration capacity. They can regenerate any amputated head or foot, and when dissociated into single cells, even intact animals will regenerate from reaggregates. This extensive regeneration capacity is mediated by epithelial stem cells, and it is based on the restoration of a signaling center, i.e., an organizer. Organizers secrete growth factors that act as long-range regulators in axis formation and cell differentiation. In Hydra, Wnt and TGF-beta/Bmp signaling pathways are transcriptionally up-regulated early during head regeneration and also define the Hydra head organizer created by de novo pattern formation in aggregates. The signaling molecules identified in Cnidarian regeneration also act in early embryogenesis of higher animals. We suppose that they represent a core network of molecular interactions, which could explain at least some of the mechanisms underlying regeneration in vertebrates. PMID- 12557205 TI - Regeneration or scarring: an immunologic perspective. AB - Complete regeneration of complex tissues and organs is usually precluded by fibrotic reactions that lead to scarring. Fish, salamanders, and larval anurans are among the few vertebrates capable of regenerating lost appendages, and this process seems to recapitulate ontogenic development of the structure in most respects. Recent work has revealed a capacity for excellent regeneration in certain mammalian tissues: embryonic or fetal skin and the ear of the MRL mouse. Analyses of these two systems suggest that processes of regenerative growth and patterning for the formation of new structures such as hair follicles may involve modulation of the inflammatory response to the injury in a way that reduces fibrosis and formation of scar tissue. We review evidence that this modulation includes changes in cytokine signaling and may involve properties of the extracellular matrix mediated by factors that include hyaluronic acid and "anti adhesive substrates" such as tenascin-C. New studies and classic work on the capacity for limb regeneration in amphibians are then reviewed, focusing on the loss of this ability in prometamorphic anuran hindlimbs and the view that changing properties of the immune system may also underlie the declining regenerative potential in this system. Finally, we review recent work in comparative and developmental immunology, which raises the possibility that phylogenetic changes in regenerative capacity may be the result of evolutionary changes in cellular activities of the immune system. PMID- 12557206 TI - Regeneration of the urodele limb: a review. AB - Urodele amphibians have been widely used for studies of limb regeneration. In this article, we review studies on blastema cell proliferation and propose a model of blastemal self-organization and patterning. The model is based on local cell interactions that intercalate positional identities within circumferential and proximodistal boundaries that outline the regenerate. The positional identities created by the intercalation process appear to be reflected in the molecular composition of the cell surface. Transcription factors and signaling molecules involved in patterning are discussed within the context of the boundary/intercalation model. PMID- 12557207 TI - Urodele spinal cord regeneration and related processes. AB - Urodele amphibians, newts and salamanders, can regenerate lesioned spinal cord at any stage of the life cycle and are the only tetrapod vertebrates that regenerate spinal cord completely as adults. The ependymal cells play a key role in this process in both gap replacement and caudal regeneration. The ependymal response helps to produce a different response to neural injury compared with mammalian neural injury. The regenerating urodele cord produces new neurons as well as supporting axonal regrowth. It is not yet clear to what extent urodele spinal cord regeneration recapitulates embryonic anteroposterior and dorsoventral patterning gene expression to achieve functional reconstruction. The source of axial patterning signals in regeneration would be substantially different from those in developing tissue, perhaps with signals propagated from the stump tissue. Examination of the effects of fibroblast growth factor and epidermal growth factor on ependymal cells in vivo and in vitro suggest a connection with neural stem cell behavior as described in developing and mature mammalian central nervous system. This review coordinates the urodele regeneration literature with axial patterning, stem cell, and neural injury literature from other systems to describe our current understanding and assess the gaps in our knowledge about urodele spinal cord regeneration. PMID- 12557208 TI - Intercalary regeneration in planarians. AB - How can a planarian regenerate its entire body from a small portion of its body? Neoblasts, the totipotent stem cells of planarian, are assumed to be able to produce all missing cell types. However, we do not know how the cell fate of these cells is controlled during regeneration. Our recent studies with molecular markers suggest that intercalary regeneration is the fundamental principle in planarian regeneration. Here, we introduce the intercalation induced by ectopic grafting along the anteroposterior (A-P), dorsoventral (D-V), and left-right (L R) axes. Blastema formation is evoked by ectopic D-V interactions after wound closure. Intercalation between the blastema and stump induces rearrangement of the positional identities along the A-P axis. Consequently, totipotent stem cells change their differentiation patterns according to the newly rearranged positional identities along the A-P, D-V, and L-R axes. According to the classic view, the blastema is regarded as the place where undifferentiated cells accumulate and regenerative events occur. Here, we propose a new interpretation, i.e., that the blastema may work as a signaling center inducing intercalary regeneration. Also, the roles of molecules and genes involved in intercalary regeneration are discussed. PMID- 12557209 TI - Identification of genes induced in regenerating Xenopus tadpole tails by using the differential display method. AB - To identify candidate gene(s) involved in the tail regeneration of Xenopus laevis tadpoles, we used the differential display method to isolate four genes (clones 1, 2, 13a, and 13b) whose expression is induced in regenerating tadpole tails. Among them, clones 13a and 13b were found to encode the Xenopus homologues of the alpha1 chain of type XVIII collagen and neuronal pentraxin I, respectively. Expression of clone 2 and neuronal pentraxin I genes increased dramatically in the blastema 3 days after amputation, whereas that for the clone 1 and type XVIII collagen genes was induced gradually after amputation. In situ hybridization revealed that the neuronal pentraxin I gene is expressed specifically in the regenerating tail epidermis but not in the normal tail epidermis or the most distal margin of the tail blastema, suggesting that it has a tissue-inductive role in tail regeneration. Expression of the four genes was induced in the limb and in the tail blastema, suggesting that they are involved in the regeneration of both organs. Finally, expression of clone 2 and neuronal pentraxin I genes was scarce during embryonic stages in comparison to the tail blastema, suggesting that their main functions are in organ regeneration. Our results demonstrate unique features of spatial and temporal gene expression patterns during Xenopus tadpole tail regeneration. PMID- 12557210 TI - Allometric scaling and proportion regulation in the freshwater planarian Schmidtea mediterranea. AB - The regulation of scale and proportion in living organisms is an intriguing and enduring problem of biology. Regulatory mechanisms for controlling body size and proportion are clearly illustrated by the regeneration of missing body parts after amputation, in which the newly regenerated tissues ultimately attain a size that is anatomically congruent with the size of the rest of the organism. Understanding the molecular processes underpinning scaling would have deep consequences for our comprehension of tissue regeneration, developmental ontogeny, growth, and evolution. Although many theories have been put forward to explain this process, it is interesting that no satisfactory mechanistic explanation is currently available to explain scalar relationships. We chose to investigate the freshwater planarian, a commonly used model system for the study of metazoan regeneration, to delineate a strategy for the molecular dissection of scale and proportion mechanisms in metazoans. Here, we report on the cloning and discrete expression pattern of a novel planarian gene, which shares homology with the DEG/ENaC super-family of sodium channels. We have named H.112.3c cintillo ("head ribbon" in Spanish) and present a strategy for using the expression of this gene to monitor scale and proportion regulation during regeneration, growth and degrowth in the freshwater planarian Schmidtea mediterranea. PMID- 12557211 TI - Mediolateral intercalation in planarians revealed by grafting experiments. AB - We investigated how planarians organize their left-right axis by using ectopic grafting. Planarians have three body axes: anteroposterior (A-P), dorsoventral (D V), and left-right (L-R). When a small piece is implanted into an ectopic region along the A-P and D-V axes, intercalary structures are always formed to compensate for positional gaps. There are two hypotheses regarding L-R axis formation in this organism: first, that the left and right sides of the animal may be recognized as different parts, and L-R intercalation can induce midline structures (asymmetry hypothesis); second, that both sides may have symmetrical positional values, and mediolateral (M-L) intercalation creates positional values along the L-R axis (symmetry hypothesis). We performed ectopic grafting experiments in the head region of the planarian, Dugesia japonica, to examine these hypotheses. A left lateral fragment containing a left auricle was implanted into the medial region of the host. Ectopic structures were always formed only on the left side of the graft, where lateral tissues abutted onto the medial tissues. However, no morphologic change was induced on the right side of the graft, where left-sided tissues faced onto right-sided tissues. Molecular marker analyses indicated that ectopic structures formed on the left side of the graft were induced by M-L intercalation, supporting the "symmetry hypothesis." When the midline tissues were implanted into a lateral region, they induced a complete ectopic head, demonstrating that M-L intercalation may be sufficient to establish the L-R axis in planarians. PMID- 12557212 TI - Recruitment of postmitotic neurons into the regenerating spinal cord of urodeles. AB - By using fluorescent tracers, we have investigated the origin of the cells that form the regenerating spinal cord after tail amputation in urodele amphibians. We show that spinal cord cells immediately adjacent to the amputation plane die and are removed by phagocytic cells. Spinal cells just anterior to these dying cells are destined to make the majority of the regenerating cord. The largest contribution is likely to come from the radial ependymal cells, but we also demonstrate that postmitotic neurons in this location can translocate into the regenerating cord. These neurons integrate into the regenerate structure and survive for at least 4 weeks. We find no evidence that these translocated neurons dedifferentiate and divide during this regeneration process. We discuss the possibility that these neurons survive long term in the regenerate cord and become part of the functional neuronal circuitry. PMID- 12557213 TI - Regeneration-specific expression pattern of three posterior Hox genes. AB - Homeobox genes encode positional information during primary and secondary axis formation during development. For this reason, the Hox genes have attracted attention in regeneration research as well. At early stages of regeneration, Hox genes have been implicated in wound healing and the dedifferentiation process and at later stages in the patterning of the blastema. We studied the expression of three Abdominal B-type Hox genes in Xenopus: XHoxc10, XHoxa13, and XHoxd13 during normal limb development and during regeneration of limbs and tails. We compared their expression with nonregenerating and with wounded limbs and tails, respectively. We show that the temporal and spatial control of these three Hox genes in blastemas differs from normal development. All three are specific to regeneration, XHoxc10 is up-regulated at the right time and at the site where cells dedifferentiate and undifferentiated cells are recruited, whereas XHoxa13 is reexpressed slightly later in regeneration, when the blastemal cells proliferate and remains on during patterning of the blastema. XHoxd13 is not expressed until relatively late and appears to be involved only in patterning of the blastema. PMID- 12557214 TI - Role for keratins 6 and 17 during wound closure in embryonic mouse skin. AB - Injury to adult skin triggers a response designed to restore its vital barrier function. A conserved aspect of this response is a rapid switch in gene expression whereby the type II keratin 6 (K6) and type I keratins 16 and 17 (K16, K17) are induced in epithelial cells at the wound edge. This induction occurs at the expense of the keratins normally expressed during terminal differentiation and correlates with the activation of epithelial cells at the wound edge, ahead of their migration into the wound site. Here, we show that the capacity to enact this switch is already acquired in E11.5 stage mouse embryos. Such early timing is well ahead of the onset of differentiation-specific gene expression (approximately E13.5) and the acquisition of barrier formation by developing epidermis (approximately E16.5). Induction of K6, K16, and K17 correlates with changes in the morphology of epithelial cells at the wound edge. The closure of embryonic wounds is significantly delayed in K17 null embryos, but not embryos null for K6. These observations significantly extend the correlation between K6, K16, and K17 expression and epithelial wound closure, and provide direct evidence that expression of these keratins, K17 in particular, is important for the timeliness of this process. PMID- 12557215 TI - Unique expression patterns of matrix metalloproteinases in regenerating newt limbs. AB - The process of regeneration of urodele limbs includes a drastic remodeling of extracellular matrices (ECMs) that is induced by matrix metalloproteinases (MMPs) and is thought to be one of the triggers of the regeneration. We studied this remodeling in limbs of Japanese newt, Cynops pyrrhogaster, by using five genes of newt MMPs (nMMPs) as probes: nMMP9, nMMP3/10-a, nMMP3/10-b, and nMMP13 that had been characterized previously, and nMMPe that was newly cloned in the present study. nMMPe was 502 amino acid residues long and showed a low homology to other known vertebrate MMPs. Reverse transcriptase-polymerase chain reactions analysis localized the transcript of nMMPe in the apical epidermal cap (AEC) and the non blastemal wound epidermis but not in the blastemal mesenchyme or the normal epidermis. Northern blot analysis localized the transcripts of nMMP9, nMMP3/10-a, and nMMP13 in the bone of regenerating limbs, whereas those of nMMP3/10-b in AEC. mRNA in situ hybridization experiments identified the nMMP-expressing cells. nMMP9 gene was strongly expressed in chondrocytes of the cartilage of epiphysis. Of interest, basal cells of AEC, but not those of the normal skin, expressed nMMP3/10-b intensely. Immunohistochemical analysis showed that the nMMP9 proteins synthesized by chondrocytes were secreted and distributed widely in the basement membrane of bone and ECMs of the amputation plane. These nMMPs characterized in the present study might cooperatively work to remodel ECMs of regenerating limbs. PMID- 12557216 TI - Matrix metalloproteinase activity correlates with blastema formation in the regenerating MRL mouse ear hole model. AB - The MRL mouse was proposed as a model of mammalian regeneration because it can close ear holes completely with the restoration of normal tissue. This regeneration process involves the formation of a blastema during healing, the re appearance of cartilage and hair follicles, and healing without scarring. Such a process requires extensive tissue remodeling. To characterize differences in ear wounding responses between regenerating and nonregenerating mice, we examined and compared the extracellular matrix remodeling and the matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) response in the MRL and C57BL/6 mouse strains after injury. We found a correlation between the MRL's ability to break down the basement membrane, form a blastema, and close ear hole wounds and an inflammatory response with neutrophils and macrophages seen in the ear after injury. These cells were positive for MMP-2 and MMP-9 as well as TIMP-2 and TIMP-3. Clear differences between the MRL and B6 response to injury were seen that could explain the differences in healing and blastema formation in the MRL and lack of it in the B6 mice. This finding was further supported by enzyme activity as determined by gelatin zymography. PMID- 12557218 TI - Identification of genes expressed during Xenopus laevis limb regeneration by using subtractive hybridization. AB - Suppression polymerase chain reaction-based subtractive hybridization was used to identify genes that are expressed during Xenopus laevis hindlimb regeneration. Subtractions were done by using RNAs extracted from the regeneration-competent stage (stage 53) and regeneration-incompetent stage (stage 59) of limb development. Forward and reverse subtractions were done between stage 53 7-day blastema and stage 53 contralateral limb (competent stage), stage 59 7-day pseudoblastema and stage 59 contralateral limb (incompetent stage), and stage 53 7-day blastema and stage 59 7-day pseudoblastema. Several thousand clones were analyzed from the various subtracted libraries, either by random selection and sequencing (1,920) or by screening subtracted cDNA clones (6,150), arrayed on nylon membranes, with tissue-specific probes. Several hundred clones were identified from the array screens whose expression levels were at least twofold higher in experimental tissue vs. control tissue (e.g., blastema vs. limb) and selected for sequencing. In addition, primers were designed to assay several of the randomly selected clones and used to assess the level of expression of these genes during regeneration and normal limb development. Approximately half of the selected clones were differentially expressed, as expected, including several that demonstrate blastema-specific enhancement of expression. Three distinct categories of expression were identified in our screens: (1) clones that are expressed in both regeneration-competent blastemas and -incompetent pseudoblastemas, (2) clones that are expressed at highest levels in regeneration competent blastemas, and (3) clones that are expressed at highest levels in regeneration-incompetent pseudoblastemas. Characterizing the role of each of these three categories of genes will be important in furthering our understanding of the process of tissue regeneration. PMID- 12557217 TI - Regeneration of the ear after wounding in different mouse strains is dependent on the severity of wound trauma. AB - The replacement and restoration of tissue mass after organ damage or injury in adult higher vertebrates is critical to the architecture and function of the organ. If replacement occurs with scar tissue, this often results in adverse effects on function and growth as well as an undesirable cosmetic appearance. However, certain mammals, such as the MRL/MpJ mouse, have shown a restricted capacity for regeneration, rather than scar tissue formation, after an excisional ear punch wound. To investigate the changes in tissue architecture leading to ear wound closure, initial ear wounding studies with a 2-mm clinical biopsy punch were performed on MRL/MpJ mice, by using C57BL/6 mice as a nonregenerative control strain. In contrast to previously reported studies on mouse ear regeneration, we observed that C57BL/6 mice in fact showed a limited regenerative capacity. One explanation for this difference could be attributed to the method of wounding used; both previous studies on mouse ear regeneration used a thumb punch, whereas our approach was to use a clinical biopsy punch. This approach led us to further investigate whether the severity of trauma applied influenced the rate of wound healing. We, therefore, compared the effects of the sharp clinical biopsy punch with that of a cruder thumb punch, and introduced a third strain of mouse, Balb/c, known to be a slow-healing strain. A new method to quantify ear punch hole closure was developed and a histologic investigation conducted up to 4 months after wounding. Image analysis data showed a reduction in original ear wound area of 85% in MRL/MpJ mice at 4 weeks and of 91.7% over 4 months by using a biopsy punch. In contrast, the crude thumb punch methodology resulted in an increase in wound area of up to 58% in Balb/c ears; thought to be due to increased necrosis of the wound site. All biopsy-punched wound areas plateaued in healing between days 28 and 112. Only 5 of 80 MRL/MpJ mouse ears showed no residual holes macroscopically after 28 days. Histologically, all strains of mice healed their ear wounds in a similar manner involving re-epithelialization, blastema-like formation, dermal extension, blood vessel formation, chondrogenesis, folliculogenesis, and skeletal muscle and fat differentiation. However, all regenerative features were more pronounced and accelerated in MRL/MpJ mice when compared with C57BL/6 and Balb/c biopsy-punched mouse ears. PMID- 12557219 TI - Bone formation during forelimb regeneration: a microtomography (microCT) analysis. AB - In our study of bone regeneration in the forelimbs of mature newts (Notophthalmus viridescens), we used noninvasive X-ray microtomography (microCT) to image regenerating limbs from 37 to 85 days and matching (contralateral) controls. We compared the patterns of regenerated and existing (nonregenerated) bone, investigating in particular the onset of mineralization of specific bones, the level of mineral present, and the lengths of the different bones. Overall, we find that the missing limb skeletal elements are restored in a proximal-to-distal direction, which reiterates the developmental patterning program. However, in contrast to this proximal-distal sequence, the portion of the humerus distal to the amputation site fails to ossify in synchrony with the regenerating radius and ulna. This finding suggests that the replacement of cartilage with mineralized bone close to the amputation site is delayed with respect to other regenerating skeletal elements. PMID- 12557220 TI - Electroporation as a tool to study in vivo spinal cord regeneration. AB - Tailed amphibians such as axolotls and newts have the unique ability to fully regenerate a functional spinal cord throughout life. Where the cells come from and how they form the new structure is still poorly understood. Here, we describe the development of a technique that allows the visualization of cells in the living animal during spinal cord regeneration. A microelectrode needle is inserted into the lumen of the spinal cord and short rapid pulses are applied to transfer the plasmids encoding the green or red fluorescent proteins into ependymal cells close to the plane of amputation. The use of small, transparent axolotls permits imaging with epifluorescence and differential interference contrast microscopy to track the transfected cells as they contribute to the spinal cord. This technique promises to be useful in understanding how neural progenitors are recruited to the regenerating spinal cord and opens up the possibility of testing gene function during this process. PMID- 12557221 TI - Lymphoma- and leukemia-associated chromosomal translocations in healthy individuals. AB - Chromosomal translocations (CTs) are hallmark mutations of hematopoietic malignancy that result in the deregulated expression of oncogenes or the generation of novel fusion genes. The polymerase chain reaction (PCR) can be used to detect illegitimate recombinations of genomic DNA sequences as a more sensitive assay than cytogenetics for determining the presence of CTs. Both direct DNA-PCR and reverse transcriptase-PCR were used to examine healthy individuals for lymphoma- and leukemia-associated CTs. Two oncogene-activating CTs [t(14;18)(q32;q21) and t(8;14)(q24;q32)] and one fusion-gene CT [t(2;5)(p23;q35)] from lymphomas and five fusion-gene CTs from leukemia [t(9;22)(q34;q11), t(4;11)(q21;q23), t(15;17)(q22;q11), t(12;21)(p13;q22), t(8;21)(q22;q22)] were detected in such studies. The biological implication is that CTs associated with malignant tumors may also be found in cells that are not neoplastic. CTs are characteristic attributes of neoplastic clones but are by themselves insufficient to cause malignant transformation. A better understanding of the special biology of non-neoplastic CT-bearing cells will provide insight into their putative role as tumor precursors. Prospective epidemiological studies are needed to determine whether such cells in healthy individuals may, in some instances, become clonogenic founders of lymphoma or leukemia. PMID- 12557222 TI - Induction of tenascin-C by tumor-specific EWS-ETS fusion genes. AB - Ewing sarcoma (ES) and peripheral primitive neuroectodermal tumors (PNETs) are associated with a chromosomal translocation resulting in a fusion of the amino terminus of EWS with the DNA-binding domain of an ETS transcription factor (most commonly FLI1 or ERG). Although previous reports suggested that these chimera proteins would act as aberrant transcription factors, their downstream targets have not been fully elucidated. To identify downstream targets of these EWS-ETS fusion proteins, we introduced EWS-ETS fusion constructs into a human fibrosarcoma cell line, HT-1080, by retroviral transduction. Here we report that Tenascin-C (TNC) is induced to a significantly higher level in cells expressing EWS-ETSs than in cells expressing normal ETSs. Furthermore, through use of an antisense cDNA expression vector we show that expression of endogenous TNC mRNA and protein were reduced coordinately with attenuation of EWS-FLI1 fusion protein expression. A chromatin immunoprecipitation assay showed direct interaction between the TNC promoter and the EWS-FLI1 fusion protein in vivo. In addition, a luciferase reporter assay revealed that EWS-ETSs upregulated the TNC gene through four ETS binding sites in the TNC promoter. High levels of TNC expression were observed in a subset of ES cell lines (3 of 6) and primary tumors (4 of 6). Together with previous studies showing that TNC expression is involved in the invasive and malignant phenotype of several tumor types, our data suggest that the oncogenic effect of EWS-ETS may be mediated in part by upregulating of TNC expression. PMID- 12557223 TI - Comprehensive molecular cytogenetic characterization of cervical cancer cell lines. AB - We applied a combination of molecular cytogenetic methods, including comparative genomic hybridization (CGH), spectral karyotyping (SKY), and fluorescence in situ hybridization (FISH), to characterize the genetic aberrations in eight widely used cervical cancer (CC) cell lines. CGH identified the most frequent chromosomal losses including 2q, 3p, 4q, 6q, 8p, 9p, 10p, 13q, and 18q; gains including 3q, 5p, 5q, 8q, 9q, 11q, 14q, 16q, 17q, and 20q; and high-level chromosomal amplification at 3q21, 7p11, 8q23-q24, 10q21, 11q13, 16q23-q24, 20q11.2, and 20q13. Several recurrent structural chromosomal rearrangements, including der(5)t(5;8)(p13;q23) and i(5)(p10); deletions affecting chromosome bands 5p11, 5q11, and 11q23; and breakpoint clusters at 2q31, 3p10, 3q25, 5p13, 5q11, 7q11.2, 7q22, 8p11.2, 8q11.2, 10p11.2, 11p11.2, 14q10, 15q10, 18q21, and 22q11.2 were identified by SKY. We detected integration of HPV16 sequences by FISH on the derivative chromosomes involving bands 18p10 and 18p11 in cell line C 4I, 2p16, 5q21, 5q23, 6q, 8q24, 10, 11p11, 15q, and 18p11 in Ca Ski, and normal chromosome 17 at 17p13 in ME-180. FISH analysis was also used further to determine the copy number changes of PIKA3CA and MYC. This comprehensive cytogenetic characterization of eight CC cell lines enhances their utility in experimental studies aimed at gene discovery and functional analysis. PMID- 12557224 TI - Rearrangements and increased expression of cyclin D1 (CCND1) in neuroblastoma. AB - Cyclin D1 regulates G1 cell cycle progression by controlling the phosphorylation of the retinoblastoma protein. This pathway is frequently deregulated in many malignancies. In neuroblastoma, however, no consistent G1 cell cycle checkpoint aberrations have been found. We examined the possible deregulation of cyclin D1 (CCND1) in this tumor. mRNA expression profiles of neuroblastoma generated by SAGE (Serial Analysis of Gene Expression) revealed a high expression of CCND1 in a subset of neuroblastoma cell lines and tumors. The CCND1 expression level can be 0.3% of the total cellular mRNA. Northern blot analysis of CCND1 expression showed a relative overexpression in 16 of 23 neuroblastoma cell lines and 10 of 15 tumor samples. In the majority of cases, the high CCND1 mRNA levels also led to high CCND1 protein levels. In the search for mechanisms causing this relative overexpression, we screened for amplifications and rearrangements of CCND1. Five amplifications were found in 202 neuroblastoma tumors and cell lines. Analysis of the 3'-UTR of CCND1 showed a rearrangement in 1 of 96 tumors. These clonal aberrations of CCND1 together with the high expression suggest a role for deregulated CCND1 activity in neuroblastoma tumorigenesis. PMID- 12557225 TI - Molecular cytogenetic characterization of Sezary syndrome. AB - Sezary syndrome (SS) is a rare form of erythrodermic cutaneous T-cell lymphoma with hematological involvement and a poor prognosis. At present little is known about the molecular pathogenesis of this malignancy. To address this issue, we analyzed 28 SS cases through the use of molecular cytogenetic techniques. Conventional cytogenetic analysis showed 12 of 28 cases with clonal chromosome abnormalities (43%). Seven cases had aberrations affecting chromosomes 1 and 17; five demonstrated rearrangement of chromosomes 10 and 14; four presented with an abnormality of 6q. Multiplex-fluorescence in situ hybridization (M-FISH) revealed complex karyotypes in 6 of 17 cases (35%), and recurrent der(1)t(1;10)(p2;q2) and der(14)t(14;15)(q;q?) translocations were each identified in two cases, and confirmed by dual-color FISH. There was an overall difference in the incidence of clonal abnormalities detected by G-banded karyotyping and M-FISH. In addition, comparative genomic hybridization studies revealed chromosome imbalances (CIs) in 9 of 20 cases (45%), with a mean DNA copy number change per sample of 1.95 +/- 2.74, and losses (mean: 1.25 +/- 1.77) more frequent than gains (mean: 0.7 +/- 1.26). The most common CIs noted were loss of 1p, followed by losses of 10/10q, 17p, and 19, and gains of 17q and 18. Furthermore, in conjunction with this study a systematic literature review was conducted, which showed a high frequency and consistent pattern of chromosome changes in SS. These findings suggest that chromosomal instability is common in SS, although there are specific chromosomal abnormalities that appear to be characteristic, and the identification of two different recurrent chromosome translocations provides the basis for further studies. PMID- 12557226 TI - Cryptic chromosomal aberrations leading to an AML1/ETO rearrangement are frequently caused by small insertions. AB - The translocation t(8;21)(q22;q22), which results in the fusion of the AML1 (RUNX1) and ETO (CBFA2T1) genes, is a recurrent aberration in acute myeloid leukemia (AML), preferentially correlated with FAB M2, and has the highest incidence in childhood AML. Because of the favorable prognosis, the evidence of the t(8;21) or the AML1/ETO fusion gene is mandatory in most of the therapy trials, allowing the stratification of the patients to the correct risk group in terms of treatment. Here we present six out of 59 children with AML who were positive for AML1/ETO by RT-PCR, but showed no evidence of the classical t(8;21)(q22;q22) by conventional cytogenetics. Because of the discrepancies between molecular and cytogenetic analyses, these six patients were further investigated by fluorescence in situ hybridization analysis. Small hidden interstitial insertions resulting in an AML1/ETO rearrangement were detected in five (8.5%) of the 59 patients, whereas the sixth patient showed a cryptic three way translocation. The insertions could be characterized as ins(21;8) in three patients and ins(8;21) in the remaining two. Additionally, three of the patients showed secondary chromosome aberrations leading to a higher complexity of the karyotype. In conclusion, the combination of more than one standard technique in the analysis of AML1/ETO is useful to reveal the overall frequency of cryptic chromosome rearrangements and permits a better understanding of the mechanisms involved in the generation of this fusion gene. PMID- 12557227 TI - Serrated adenomas and mixed polyposis caused by a splice acceptor deletion in the mouse Smad4 gene. AB - Serrated adenomas, hyperplastic polyps, and admixed hyperplastic/adenomatous polyps form a distinct group of colorectal tumors, the molecular genetic basis of which is still poorly understood. We describe a novel mouse model for serrated adenomas and mixed polyposis, here referred to as Sad (serrated adenomas), caused by a spontaneously risen splice site mutation in the murine Smad4 gene. The Sad chromosomal region was identified by genetic linkage and loss of heterozygosity (LOH) analysis. Subsequently, several candidate genes were investigated by expression and mutation analysis. By use of genetic linkage and LOH analysis, we mapped the Sad candidate to mouse chromosome 18, 44-48 cM, syntenic to human chromosome band 18q21. Within this chromosomal interval, the Smad2, Smad4, and Smad7 genes were analyzed for the presence of a disease-causing mutation in affected animals. A single nucleotide (nt) deletion was identified in the intron 5/exon 6 splice acceptor site of the Smad4 gene. The single base deletion results in a frameshift and an early termination codon through activation of a cryptic splice site 4 nt downstream in exon 6. The resulting mRNA is unstable, and the Sad mutation is thus likely to represent a null allele. Identification of a Smad4 mutation in the Sad mouse model provides further support for the involvement of the Smad genes, and thus the TGFB pathway, in the serrated/hyperplastic route to colorectal cancer. PMID- 12557228 TI - SIAH1 inactivation correlates with tumor progression in hepatocellular carcinomas. AB - Accumulation of loss of heterozygosity (LOH) on chromosome 16 is frequently observed in human hepatocellular carcinomas (HCCs). To identify tumor-suppressor genes (TSGs) involved in hepatocarcinogenesis, we performed deletion mapping of chromosome 16 in 59 HCCs. Three commonly deleted regions, located in 16q12.1, 16q22.1, and 16q24.2, were observed. Because there has been no study on LOH at locus 16q12.1 in HCCs, we focused on this region. By searching the Human Genome Database at the National Center for Biotechnology Information web site, we identified 14 known genes in 16q12.1 as TSG candidates. Among these, the expression of SIAH1 was markedly downregulated in HCCs, and inactivation of SIAH1 expression was associated with LOH at 16q12.1. A mutation analysis of SIAH1 revealed no somatic mutations, but one single nucleotide polymorphism was found among the 35 HCCs investigated. Subsequently, we evaluated the relation between SIAH1 expression, confirmed by semiquantitative RT-PCR, and clinicopathological parameters in HCCs. SIAH1 was significantly downregulated in advanced HCCs, including poorly differentiated tumors, larger tumors, and tumors in advanced stages. These findings suggest that inactivation of SIAH1 plays an important role in HCC progression. PMID- 12557229 TI - Chromosome imbalances in thyroid follicular neoplasms: a comparison between follicular adenomas and carcinomas. AB - The underlying genetic events associated with follicular thyroid tumorigenesis are still ill defined. In this study, we performed a screening for chromosome imbalances by comparative genomic hybridization (CGH) in a group of 12 follicular adenomas (FAs) and 20 follicular carcinomas (FTCs) previously characterized by conventional cytogenetics and flow cytometry analysis. In general, a great similarity was observed between the CGH profiles of the FAs and FTCs. In both benign and malignant tumors, a combination of gains affecting 5, 7, 12, 17, 19, and 20 was observed. Chromosome 7 was the most frequently affected chromosome, with three regions of consensus gains: 7p11-12, 7q11.3-q21, and 7q31. Recurrent gains of chromosomes 5 and 12 involved 5p11, 5p15, 5q13-q22, 5q21-q23, 12p11, and 12q11-q12. DNA sequence losses were also observed in both tumor groups. Chromosomal arms deleted in at least five of the neoplasms were (in order of frequency): in adenomas, 15q, 2p/2q, 3q, 6p/6q, 11q, and 22q; and in FTCs, 3p, 2p, 8q, 1p, 2q, 3q, 6q, 8p, 9p, 11q, 13q, 6p, and 18q. The statistical evaluation of the CGH data demonstrated that 15q loss was significantly associated with FA. Two regions of minimal common loss were defined by CGH at 15: 15q11-q21 and 15q26 qter. The identification of these regions provides a basis for further molecular studies. PMID- 12557230 TI - Genetic markers useful for distinguishing between organ-confined and locally advanced prostate cancer. AB - Prostate cancer is one of the most commonly diagnosed cancers and the second leading cause of cancer deaths among men in the United States. In this study, we performed comparative genomic hybridization (CGH) on 45 primary prostate adenocarcinomas to determine genetic markers that could be useful for distinguishing between organ-confined and locally advanced prostate cancer. Of these tumors, 24 were pT2 stage, 21 were pT3b; 20 had low Gleason scores (GS), 25 had high GS. The most common chromosomal alterations in all 45 tumors included losses on 8p (57.8%), 10q21-->qter (40.0%), 16q (35.6%), 11q21-->qter (28.9%), 16p (22.2%), 6q22-->24 (22.2%), 10p (20.0%), 5q31-->qter (17.8%), 6p (17.8%), 15q22-->qter (15.6%), and 17p (15.6%) as well as gains on 7cen-->p14 (20.0%), 7cen-->q22 (20.0%), and Xcen-->q21 (17.8%). Contingency table analysis showed that losses of 8p, 10q25-->qter, 6p21, 6q24-->qter, and 15q22-->qter were significantly increased in frequency (P < 0.05) with increasing stage and/or GS. A model was created following multivariate logistic regression analysis that was predictive of tumor stage in approximately 90% of the tumors studied. This model suggests that loss of 8p is the most valuable predictor of stage. These findings suggest that chromosomal regions identified in this study may be useful for distinguishing between organ-confined and locally advanced prostate tumors. PMID- 12557231 TI - Breakpoints at 1p36.3 in three MDS/AML(M4) patients with t(1;3)(p36;q21) occur in the first intron and in the 5' region of MEL1. AB - The recurrent translocation t(1;3)(p36;q21) is associated with myelodysplastic syndrome (MDS)/acute myelogenous leukemia (AML) characterized by trilineage dysplasia, especially dysmegakaryopoiesis and a poor prognosis. Recently, the two genes involved in this translocation have been identified: the MEL1 gene at 1p36.3, and the RPN1 gene at 3q21. The breakpoint in RPN1 is centromeric to the breakpoint cluster region of the inv(3) abnormality. Because the MEL1 transcript is detected only in leukemic cells with t(1;3)(p36;q21), ectopic expression of MEL1 driven by RPN1 at 3q21 is thought to contribute to the pathogenesis of t(1;3)(p36;q21) leukemia. However, the precise breakpoint in the patients has not yet been identified. With fluorescence in situ hybridization analysis by use of BAC/PAC probes, we identified the breakpoint at 1p36.3 in three MDS/AML patients with t(1;3)(p36;q21): within the first intron of the MEL1 gene (one patient) or within a 29-kb region located in the 5' region of MEL1 (two other patients). We detected several sizes of MEL1 transcript in two patients including the first patient, although we have not yet clarified whether MEL1 transcripts were different among the patients and whether a truncated MEL1 transcript was expressed in the first patient. This patient showed an unusual clinical profile, repeating progression to overt leukemia and conversion to MDS three times during the 29-month survival period, which might be related to a different molecular mechanism in this patient. PMID- 12557232 TI - The use of the resonant mirror biosensor to detect point mutations, as demonstrated with synthetic oligonucleotides. AB - The possibility of using the resonant mirror biosensor to detect point substitutions in oligonucleotides was demonstrated with a fragment of the Helicobacter pylori 23S rRNA gene, point mutations in which are responsible for clarythromycin resistance. Conditions were optimized for the interaction of a probe immobilized on the sensing surface with targets containing various nucleotide substitutions. A probe allowing reliable discrimination of mutant targets was selected. The mismatch position in the probe was shown to affect the kinetic parameters (response) of hybridization with mutant targets, reporting not only the position, but also the character (G or C) of a substitution. PMID- 12557233 TI - Delineating protein-protein interactions via biomolecular interaction analysis mass spectrometry. AB - The utility of biomolecular interaction analysis-mass spectrometry (BIA/MS) in screening for protein-protein interactions was explored in this work. Experiments were performed in which proteins served as ligands for screening of possible interactions with other proteins from human plasma and urine. The proteins utilized were beta-2-microglobulin, cystatin C (cysC), retinol binding protein (RBP), transthyretin (TTR), alpha-1-microglobulin, C-reactive protein, transferrin and papain. The immobilization of functionally active proteins was confirmed via interactions with antibodies to the corresponding proteins. Various dilutions of human urine and plasma were injected over the protein-derivatized surfaces. It was observed that the urine injections generally yielded smaller SPR responses than those observed after the plasma injections. The BIA/MS experiments did not reveal novel protein-protein interactions, although several established interactions (such as those between RBP and TTR, and cysC and papain) were validated. Few protein ligand deficiencies (such as truncations) leading to false negative and false positive BIA/MS results were also discovered. PMID- 12557234 TI - Design and use of multi-affinity surfaces in biomolecular interaction analysis mass spectrometry (BIA/MS): a step toward the design of SPR/MS arrays. AB - The feasibility of multi-affinity ligand surfaces in biomolecular interaction analysis-mass spectrometry (BIA/MS) was explored in this work. Multi-protein affinity surfaces were constructed by utilizing antibodies to beta-2 microglobulin, cystatin C, retinol binding protein, transthyretin, serum amyloid P and C-reactive protein. In the initial experiments, all six antibodies were immobilized on a single site (flow cell) on the sensor chip surface, followed by verification of the surface activity via separate injections of purified proteins. After an injection of diluted human plasma aliquot over the antibodies derivatized surfaces, and subsequent MALDI-TOF MS analysis, signals representing five out of the six targeted proteins were observed in the mass spectra. Further, to avoid the complexity of the spectra, the six proteins were divided into two groups (according to their molecular weight) and immobilized on two separate surfaces on a single sensor chip, followed by an injection of human plasma aliquot. The resulting mass spectra showed signals from all proteins. Also, the convolution resulting from the multiply charged ion species was eliminated. The ability to create such multi-affinity surfaces indicates that smaller-size ligand areas/spots can be employed in the BIA/MS protein interaction screening experiments, and opens up the possibilities for construction of novel multi arrayed SPR-MS platforms and methods for high-throughput parallel protein interaction investigations. PMID- 12557235 TI - BEPITOPE: predicting the location of continuous epitopes and patterns in proteins. AB - In growing need of obtaining highly specific monoclonal antibodies against novel proteins, we developed new functions implemented in the program BEPITOPE to predict continuous protein epitopes. This program not only can compute, combine, display and print prediction profiles, but also provides a list of suggested linear peptides to be synthesized. Novel facilities incorporated in BEPITOPE include the treatment of a whole genome, the search for a user-defined pattern, and the combination of prediction to pattern profiles. This latter approach is useful to remove unwanted predictions such as those including glycosylation sites. PMID- 12557236 TI - RNA aptamers specific for bovine thrombin. AB - Bovine thrombin is widely used in clinical wound healing after surgery. There is 85% homology between bovine thrombin and human thrombin, so most antibodies against bovine thrombin cross-react with human thrombin. Rare antibodies against bovine thrombin but not cross-reacting with human thrombin have been reported. RNA ligands (aptamers) have been used to bind to target molecules with sometimes higher specificity than antibodies. Here we report the isolation of aptamers specific for bovine thrombin by systematic evolution of ligands by exponential enrichment (SELEX) from an RNA pool containing a 25-nucleotide randomized region. After seven rounds of selection, two aptamers specific for bovine thrombin were identified with a K(d) of 164 and 240 nM, respectively. Significantly, these aptamers do not bind to human thrombin. Secondary structure prediction revealed potential stem-loop structures for these RNAs. Both RNA aptamers inhibit only bovine thrombin-catalyzed fibrin clot formation in vitro. Competition assay results suggested that the RNA aptamers might bind to the electropositive domain of bovine thrombin, that is, heparin-binding site, instead of fibrinogen recognition exosite. The resulting bovine-specific thrombin inhibitor might be used in some clinical applications when bovine thrombin activity needs to be contained or in research where human and bovine thrombin need to be distinguished. PMID- 12557238 TI - Kinetic analysis of the interactions between troponin C (TnC) and troponin I (TnI) binding peptides: evidence for separate binding sites for the 'structural' N-terminus and the 'regulatory' C-terminus of TnI on TnC. AB - The Ca(2+)/Mg(2+)-dependent interactions between TnC and TnI play a critical role in regulating the 'on' and 'off' states of muscle contraction as well as maintaining the structural integrity of the troponin complex in the off state. In the present study, we have investigated the binding interactions between the N terminus of TnI (residues 1-40 of skeletal TnI) and skeletal TnC in the presence of Ca(2+) ions, Mg(2+) ions and in the presence of the C-terminal regulatory region peptides: TnI(96-115), TnI(96-131) and TnI(96-139). Our results show the N terminus of TnI can bind to TnC with high affinity in the presence of Ca(2+) or Mg(2+) ions with apparent equilibrium dissociation constants of K(d(Ca(2+) ) ) = 48 nM and K(d(Mg(2+) ) ) = 29 nM. The apparent association and dissociation rate constants for the interactions were, k(on) = 4.8 x 10(5) M (-1) s(-1), 3.4 x 10(5) M (-1) s(-1) and k(off) = 2.3 x 10(-2) s(-1), 1.0 x 10(-2) s(-1) for TnC(Ca(2+)) and TnC(Mg(2+)) states, respectively. Competition studies between each of the TnI regions and TnC showed that both TnI regions can bind simultaneously to TnC while native gel electrophoresis and SEC confirmed the formation of stable ternary complexes between TnI(96-139) (or TnI(96-131)) and TnC-TnI(1-40). Further analysis of the binding interactions in the ternary complex showed the binding of the TnI regulatory region to TnC was critically dependent upon the presence of both TnC binding sites (i.e. TnI(96-115) and TnI(116-131)) and the presence of Ca(2+). Furthermore, the presence of TnI(1-40) slightly weakened the affinity of the regulatory peptides for TnC. Taken together, these results support the model for TnI-TnC interaction where the N terminus of TnI remains bound to the C-domain of TnC in the presence of high and low Ca(2+) levels while the TnI regulatory region (residues 96-139) switches in its binding interactions between the actin-tropomyosin thin filament and its own sites on the N- and C-domain of TnC at high Ca(2+) levels, thus regulating muscle contraction. PMID- 12557237 TI - Human CD30: structural implications from epitope mapping and modeling studies. AB - The human CD30 molecule is expressed transiently at very low levels on intrafollicular and perifollicular T and B cell blasts in lymphoid tissues, but is specifically upregulated on certain tumor cells, e.g. Hodgkin and Reed Sternberg (H-RS) cells. With its specific expression pattern and easy accessibility on the surface of H-RS cells CD30 is a valuable diagnostic marker and holds considerable promise as a target for in vivo immunotherapy. Knowledge of epitopes on the CD30 molecule is expected to facilitate the design of novel non-immunogenic anti-CD30 reagents. Therefore, we have mapped the epitopes of several monoclonal antibodies (mAb) applying a peptide array of overlapping CD30 derived peptides. For the mAb Ber-H2, two linear epitopes with identical sequence were found, while the mAb Ki-2 and the single chain Fv fragment R4-4 each recognized a single linear antigenic determinant, respectively. On the other hand, the mAb Ki-1 bound to a discontinuous epitope composed of two regions, one located near the N-terminus and the other near the membrane-spanning region of CD30. Using molecular modeling, it was possible to visualize the location of the epitopes on exposed loop regions of the molecule within the N-terminal domain. Finally, the results obtained with the mAb Ki-1 imply that the ends of the N- and C-terminal parts of the extracellular portion of CD30 are in close vicinity of each other, suggesting a flower-like structure for the membrane-bound homotrimeric CD30 molecule. PMID- 12557239 TI - Interaction of epitope-related and -unrelated peptides with anti-p24 (HIV-1) monoclonal antibody CB4-1 and its Fab fragment. AB - The binding of four epitope-related peptides and three library-derived, epitope unrelated peptides of different lengths (10-14 amino acids) and sequence by anti p24 (HIV-1) monoclonal antibody CB4-1 and its Fab fragment was studied by isothermal titration calorimetry. The binding constants K(A) at 25 degrees C vary between 5.1 x 10(7) M (-1) for the strongest and 1.4 x 10(5) M (-1) for the weakest binder. For each of the peptides complex formation is enthalpically driven and connected with unfavorable entropic contributions; however, the ratio of enthalpy and entropy contributions to deltaG(0) differs markedly for the individual peptides. A plot of -deltaH(0) vs -TdeltaS(0) shows a linear correlation of the data for a wide variety of experimental conditions as expected for a process with deltaC(p) much larger than deltaS(0). The dissimilarity of deltaC(p) and deltaS(0) also explains why deltaH(0) and TdeltaS(0) show similar temperature dependences resulting in relatively small changes of deltaG(0) with temperature. The heat capacity changes deltaC(p) upon antibody-peptide complex formation determined for three selected peptides vary only in a small range, indicating basic thermodynamic similarity despite different key residues interacting in the complexes. Furthermore, the comparison of van't Hoff and calorimetric enthalpies point to a non-two-state binding mechanism. Protonation effects were excluded by measurements in buffers of different ionization enthalpies. Differences in the solution conformation of the peptides as demonstrated by circular dichroic measurements do not explain different binding affinities of the peptides; specifically a high helix content in solution is not essential for high binding affinity despite the helical epitope conformation in the crystal structure of p24. PMID- 12557241 TI - Seasonal variation of the lipoidal matters and hypolipidaemic activity of the red alga Corallina officinalis L. AB - The lipoidal matters of Corallina officinalis L. showed a seasonal nonsignificant quantitative variation. However, the fatty acids revealed a relative increase in the summer and winter, while unsaponifiable matter exhibited a slight increase in the spring. The GC/MS analysis of saponifiable and unsaponifiable matter of the algal samples collected in different seasons revealed that samples collected in the spring contained a low cholesterol content and high steroidal compounds as well as high polyunsaturated fatty acids. The alcohol extract, hexane extract and fatty acid fraction of this algal sample exhibited a significant hypolipidaemic activity. Also, two biologically active fractions of hydrocarbons were isolated by CC technique from the hexane fraction of C. officinalis L. and identified by GC/MS. PMID- 12557240 TI - Plants with traditional uses and activities, relevant to the management of Alzheimer's disease and other cognitive disorders. AB - In traditional practices of medicine, numerous plants have been used to treat cognitive disorders, including neurodegenerative diseases such as Alzheimer's disease (AD) and other memory related disorders. An ethnopharmacological approach has provided leads to identifying potential new drugs from plant sources, including those for memory disorders. There are numerous drugs available in Western medicine that have been directly isolated from plants, or are derived from templates of compounds from plant sources. For example, some alkaloids from plant sources have been investigated for their potential in AD therapy, and are now in clinical use (e.g. galantamine from Galanthus nivalis L. is used in the United Kingdom). Various other plant species have shown favourable effects in AD, or pharmacological activities indicating the potential for use in AD therapy. This article reviews some of the plants and their active constituents that have been used in traditional medicine, including Ayurvedic, Chinese, European and Japanese medicine, for their reputed cognitive-enhancing and antidementia effects. Plants and their constituents with pharmacological activities that may be relevant to the treatment of cognitive disorders, including enhancement of cholinergic function in the central nervous system, anti-cholinesterase (anti ChE), antiinflammatory, antioxidant and oestrogenic effects, are discussed. PMID- 12557242 TI - Hypoglycaemic effect of Opuntia lindheimeri Englem in a diabetic pig model. AB - The hypoglycaemic activity of Opuntia lindheimeri Englem. was investigated in non diabetic (control pigs) and streptozotocin-induced diabetic pigs using an enteral (oral) route of administration. Following the administration of O. lindheimeri extract (0, 250 or 500 mg/kg body weight), blood glucose concentrations in control pigs fluctuated around initial baseline concentrations, but were not consistently affected by either the dose of O. lindheimeri or by the time following administration. In contrast, administration of O. lindheimeri extract to STZ-treated pigs resulted in both a dose- (p < 0.001) and time-dependent (p < 0.001) decrease in blood glucose concentrations. The hypoglycaemic effect of the extract was apparent within 1 h of administration, with maximal effects occurring at 4 h after administration. These results confirm the hypoglycaemic effect of O. lindheimeri extract in a diabetic pig model. In addition, given the physiological similarities of the pig to humans, this model will be of tremendous use in assessing the long-term effects of Opuntia administration on the secondary problems associated with diabetes. PMID- 12557243 TI - Hepatoprotective and antihyperliposis activities of in vitro cultured Anoectochilus formosanus. AB - The pharmacological effects of an aqueous extract of the whole plants of in vitro cultured Anoectochilus formosanus were investigated experimentally for hepatoprotective and antihyperliposis activities. The extract showed significant antihepatotoxic activity against carbon tetrachloride induced cytotoxicity in primary cultured rat hepatocytes. In an assay for antihyperliposis using aurothioglucose-induced obese mice, the extract suppressed significantly the increase in the weights of body and liver, ameliorated triglyceride levels in the liver and serum, and also significantly reduced the deposition of adipose tissue. PMID- 12557244 TI - In vitro immunomodulatory activity of flavonoid glycosides from Urtica dioica L. AB - The major compounds isolated from the methanolic extract of the aerial parts of Urtica dioica L. were determined as quercetin-3-O-rutinoside (1). kaempherol-3-O rutinoside (2). and isorhamnetin-3-O-glucoside (3). by chromatographic, chemical (acidic hydrolysis) and spectral (UV, IR, (1)H-NMR, (13)C-NMR) methods. Their immunomodulatory activities were studied in vitro by chemotaxis (Boyden Migration Chamber) and intracellular killing activity (NBT reduction) tests. Compounds 1, 2, 3 and the total flavonoid fraction were determined to have significant chemotactic effects in 4, 8, 16 microg doses. According to the results of the NBT reduction test, all flavonoid glycosides showed high intracellular killing activity. The results of both assays confirmed the immunostimulatory activity of the flavonoid fraction and the isolated flavonoid glycosides on neutrophils suggesting that they could possibly be useful for treating patients suffering from neutrophil function deficiency and chronic granulomatous diseases. PMID- 12557245 TI - Anticonvulsant effect of FS-1 subfraction isolated from roots of Delphinim denudatum on hippocampal pyramidal neurons. AB - The effects were investigated of a partially purified subfraction (FS-1) isolated from Delphinium denudatum on sustained repetitive firing (SRF) of cultured neonatal rat hippocampal pyramidal neurons. The blockade of sustained repetitive firing is one of the basic mechanisms of antiepileptic drugs at the cellular level. Using the whole cell current-clamp technique, sustained repetitive firing was elicited in pyramidal neurons under study by a depolarizing pulse of 500 ms duration, 0.3 Hz and 0.1-0.6 nA current strength. FS-1 (0.01-0.06 mg/mL) reduced the number of action potentials per pulse in a dose-dependent manner until no action potentials were elicited for the remainder of the pulse. There was a corresponding use-dependent reduction in amplitude and Vmax of action potentials. The Vmax of action potential 1 exhibited a dose-dependent reduction. At a dose of 0.06 mg/mL FS-1 reduced Vmax to 29%-38% and amplitude to 16%-20 % of the control values. The blockade of sustained repetitive firing by FS-1 was reversed by hyperpolarization of the membrane potential (-65 to -75 mV) while depolarization of the membrane potential (-53 mV to -48 mV) potentiated the block. The results suggest that FS-1 blocks sustained repetitive firing in hippocampal neurons in a use-dependent and voltage-dependent manner similar to the prototype anticonvulsant drug, phenytoin. However, unlike phenytoin, which binds preferably to the inactive state, the compounds present in FS-1 also interacted with the resting state of the Na+ channels by reducing Vmax of action potential 1. The results indicate that the partially purified FS-1 subfraction of Delphinium denudatum contains a potent anticonvulsant compound. PMID- 12557246 TI - Antioxidant effect of polyamines on erythrocyte cell membrane lipoperoxidation after free-radical damage. AB - This in vitro study determined the effect of three doses each (100, 500 and 1000 microm) of putrescine, spermidine and spermine on malondialdehyde (MDA) release in red blood cells (RBCs) from healthy individuals after hydrogen peroxide stimulation (10 mM). Twenty-two volunteers, 9 males and 13 females, aged 41.5 +/- 16.4 years, were studied. MDA was measured by thiobarbituric reaction (TBARs) and the results were calculated using epsilon = 1.56 x 10(5). The analysis of variance (ANOVA) demonstrated a statistically significant overall decrease in MDA release in the polyamine-exposed cells (p < 0.0001) when compared with unexposed cells. Individual analysis of each polyamine separately showed a 52% decrease in MDA release with added spermine and a 39.5% decrease with added spermidine (p < 0.001). No evaluable effect was found for putrescine. There was no correlation between the effect produced and the three doses of spermidine or spermine added, indicating a non dose-dependent action. PMID- 12557247 TI - Prevention of diet-induced type 2 diabetes in the C57BL/6J mouse model by an antidiabetic herbal formula. AB - Significant positive effects of an antidiabetes herbal formula (ADHF) on the prevention of progressive diet-induced type 2 diabetes (NIDDM) in the well recognized C57BL/6J mouse model were demonstrated. The results of the present studies confirm that C57BL/6J mice, through a genetic predisposition, developed NIDDM-like symptoms on a regular schedule when fed solely on a diabetes induction diet containing high fat, high simple carbohydrate and low fibre. Blood glucose levels consistent with type 2 diabetes were detected after 8 weeks on this induction diet. In contrast, no members of the prevention study groups of C57BL/6J mice, which had ADHF mixed into their diabetes induction diet, developed progressive evidence of NIDDM-like symptoms. Specific parameters observed to be significantly reduced in mice in the prevention diet groups fed ADHF mixed into the diabetes induction diet compared with the diabetes positive control groups fed diabetes induction diet only were clinical signs, blood glucose and serum insulin levels, insulin resistance and histopathological changes in major organs. This is the first report to our knowledge to show in vivo evidence for the successful prevention of progressive NIDDM-like symptoms by the addition of an anti-diabetes herbal formula to a effective diabetes induction diet. PMID- 12557248 TI - The effect of a water extract and anthocyanins of hibiscus sabdariffa L on paracetamol-induced hepatoxicity in rats. AB - We investigated the effect of the water extract of the dried flowers of Hibiscus sabdariffa L. and Hibiscus anthocyanins (HAs) (which are a group of natural pigments occurring in the dried calyx of H. sabdariffa) on paracetamol-induced hepatotoxicity in rats. The water extract was given in lieu of drinking water for 2, 3 or 4 consecutive weeks, and the HAs were given orally at doses of 50, 100 and 200 mg/Kg for five consecutive days. Paracetamol was given orally at a dose of 700 mg/Kg to induce hepatotoxicity at the end of the water extract and Has treatments. Six hours thereafter the rats were killed and their liver function evaluated biochemically and histologically. Given for 4 weeks (but not for 2 or 3 weeks) the extract significantly improved some of the liver function tests evaluated, but did not alter the histology of the paracetamol-treated rats or the pentobarbitone-induced sleeping time. At a dose of 200 mg/Kg, the hepatic histology and the biochemical indices of liver damage were restored to normal. Lower does were ineffective. Pending more evaluation for safety and efficacy, the HAs can potentially be used in mitigating paracetamol-induced hepatotoxicity. PMID- 12557249 TI - The comparative effect of novel Pelargonium essential oils and their corresponding hydrosols as antimicrobial agents in a model food system. AB - Essential oils and their corresponding hydrosols, obtained after distillation of various scented Pelargonium (Geraniaceae) leaves were assessed for their antimicrobial activity in a model food system. Both the essential oils and hydrosols were used at 1000 ppm in broccoli soup, previously inoculated with Enterobacter aerogenes (at 10(5) cfu g(-1)) and Staphylococcus aureus (at 10(4) cfu g(-1)). The results showed a complete inhibition of S. aureus in the broccoli soup by the essential oils of 'Sweet Mimosa', 'Mabel Grey', P. graveolens, 'Atomic Snowflake', 'Royal Oak', 'Attar of Roses' and a lesser effect by 'Chocolate Peppermint' and 'Clorinda'; the hydrosols, however, had a potentiating effect on the bacterial population in the food. Both extracts showed a complete inhibition of S. aureus in the Maximum Recovery Diluent (MRD). Antibacterial activity against E. aerogenes in the broccoli soup was generally very much reduced: only the essential oil of 'Mabel Grey' showed complete inhibition and virtually no reductions in colonies were seen with the other essential oils; the hydrosols again caused an increase in bacterial colonies. All the essential oils, bar Chocolate Peppermint showed complete inhibition of E. aerogenes in MRD, but the hydrosols showed no effect. The results strongly suggest that the residual hydrosols from distillation of these plant essential oils have no potential as antibacterial agents in foods, in contrast to most of the essential oils, which show potential against some micro-organisms, but only in some food systems. The problem of food component interference and its possible management is discussed. PMID- 12557250 TI - Pycnogenol inhibits the release of histamine from mast cells. AB - Oxygen derived free radicals are now increasingly regarded as a primary force of tissue destruction and also have the ability to release histamine from mast cells. Pycnogenol is an extract of the bark of French maritime pine (Pinus pinaster) containing bioflavonoids with a potent ability to scavenge free radicals. Therefore Pycnogenol was investigated for inhibition of histamine release from rat peritoneal mast cells. In addition, its effects were compared with sodium cromoglycate, a known inhibitor of histamine release from the mast cell. Rat peritoneal mast cells were isolated and purified by differential centrifugation and cells pooled from 3-4 animals were suspended at approximately 10(6) cells/mL buffered salt solution. Histamine release was induced by compound 48/80 or the calcium ionophore A-23187 and estimated from supernatant following extraction and by fluorimetric methods. Pycnogenol produced a concentration dependent inhibition of histamine release induced by the two secretagogues. Its inhibitory effect on mast cell histamine release was favourably comparable to sodium cromoglycate. PMID- 12557251 TI - Antiinvasive, antiangiogenic and antitumour activity of Ephedra sinica extract. AB - A water fraction (EW), the remaining water phase from the methanol extract of Ephedra sinica after extraction with ethyl acetate and butanol, has been investigated for its antiangiogenic, antiinvasive and antitumour activities. It was observed that EW, at 30 micro g/mL, a non-cytotoxic concentration, inhibited the tube formation induced by human umbilical venous endothelial cells and the invasion of B16F10 melanoma cells through a matrix membrane by more than 90%. At 30 mg/kg/day, the inhibitory activity of EW on the growth of a tumour mass in BDF1 mice inoculated with B16-F10 murine melanoma cells was comparable to that of adriamycin (ADR) administered at 2 mg/kg/day. From these results, it could be postulated that the antitumour activity of the water fraction might be attributed to its antiangiogenic and antiinvasive activities. PMID- 12557252 TI - Effect of alcohol extract of Achyranthes aspera Linn. on acute and subacute inflammation. AB - The antiinflammatory activity of an alcohol extract of Achyranthes aspera was tested on carrageenin-induced hind paw oedema and cotton pellet granuloma models in albino male rats. The paw volume was measured plethysmometrically at 0, 1, 2, 3, 4 and 5 h. In the subacute model cotton pellet granuloma was produced by implantation of 50 +/- 1 mg sterile cotton in the axilla under ether anaesthesia. The animals were fed with an alcohol extract at various dose levels (125, 250, 375 and 500 mg/kg). Diclofenac sodium was used as a standard drug. The alcohol extract (375 and 500 mg/kg) showed the maximum inhibition of oedema of 65.38% and 72.37% at the end of 3 h with carrageenin-induced rat paw oedema, respectively. Using a chronic test, the extract exhibited a 40.03% and 45.32% reduction in granuloma weight. PMID- 12557253 TI - Pharmacological activity of South American plants: effects on spontaneous in vivo lipid peroxidation. AB - The in vivo protective effects of methanol extracts of eight South American medicinal plants traditionally used as antiinflammatory were determined by means of spontaneous lipid peroxidation of liver tissue in rats. The production of TBARS was reduced in a dose dependent manner for A. macrocarpa (IC50 = 132 mg/kg), A. urundeuva (IC50 = 176 mg/kg), C. reticulata (IC50 = 561mg/kg) and S. obtusifolium (IC50 = 918 mg/kg). The extracts of P. peltata and U. tomentosa were only effective at a high concentration (300 mg/kg), although these values were not significant. The lyophilized latex of C. lechleri decreased the production of TBARS at a 200 mg/kg dose, although pro-oxidant effects were observed at lower doses (50 mg/kg). The extract of H. pallida was pro-oxidant at lower concentrations (50 mg/kg). PMID- 12557254 TI - Biological activities of synthetic saponins and cardiac glycosides. AB - Comparisons of the biological activities of diosgenyl (DIO) beta-D-xylopyranosyl- (1 --> 2)-alpha-D-glucopyranoside (2DXalpha) and beta-D-glucopyranosyl- (1 --> 3) beta-D-glucopyranoside (3GLbeta) or methyl glycyrrhetinate (GAM) 2DXalpha and 3GLbeta with those of the other DIO- or GAM- glycosides confirmed our assumption that both haemolytic and antifungal activities of DIO-glycosides (steroid saponins) generally parallel each other, while almost all haemolytic GAM glycosides (triterpenoid saponins) have no antifungal activity. By comparisons of digitoxigenyl (DIG) 2DXalpha, beta-D-xylopyranosyl- (1 --> 2)-beta-D glucopyranoside (2DXbeta) and 3GLbeta with the other DIG-glycosides, the previous supposition on DIG-diglycosides (cardiac diglycosides) has been developed into the assumption that those having a (1 --> 4), (1 --> 3) or (1 --> 2)beta sugar linkage show stronger anti-ATPase and antiviral activities than those with a (1 - > 6) or (1 --> 2)alpha linkage. PMID- 12557255 TI - Correction of haemorheological disturbances in myocardial infarction by diquertin and ascorbic acid. AB - In a model of the high blood viscosity syndrome, developed after myocardial infarction in rats, it was observed that a therapy of a combination of diquertin (20 mg/kg) and ascorbic acid (50 mg/kg) for a -period of 6 days, resulted in an improvement of haemorheological indices. The decrease in blood -viscosity was primarily due to an improved deformability of erythrocytes, and to some extent, due to a decrease in the content of plasma fibrinogen and erythrocyte aggregation. PMID- 12557256 TI - Downregulation of hepatitis B surface antigen expression in human hepatocellular carcinoma cell lines by HD-03, a polyherbal formulation. AB - Hepatitis B virus is associated with chronic or acute liver diseases and with hepatocellular carcinoma. In the present study, we examined the activity of HD 03, a polyherbal formulation, on two hepatitis B surface antigen (HBsAg) expressing human hepatocellular carcinoma cell lines, PLC/PRF/5 and HepG2/A2. We observed that HD-03 downregulates HBsAg expression from these cell lines. Our studies have also shown that this effect is neither due to cytotoxicity on the cell lines nor due to blockade of the release of the antigen from the cells nor due to binding of the substance with the antigen. The possible mode of its antiviral activity is explained. PMID- 12557257 TI - Effect of combination of Capsicum frutescens and Citrullus colocynthis on growth, haematological and pathophysiological parameters of rats. AB - The toxicity of diet containing 10% of Capsicum frutescens or 10% of Citrullus colocynthis fruits or their 1:1 mixture (5% + 5%) to rats treated for 6 weeks was evaluated. Body weight loss, inefficiency of feed utilization, diarrhoea, and enterohepatonephropathy characterized C. colocynthis toxicosis in rats. Despite impairment of rat's growth neither nephropathy nor diarrhoea was detected in rats fed the 10% C. frutescens diet. Feeding the mixture of C. frutescens and C. colocynthis caused more pronounced effects and death of rats. Vital organ lesions accompanied by anaemia and leucopenia were correlated with changes in serum ALP, AST and ALT activities with alterations in concentrations of total protein, albumin, urea and other serum constituents. Serum bilirubin concentration did not change. PMID- 12557259 TI - Exploring environmental causes of altered ras effects: fragmentation plus integration? AB - Mutations in ras genes are the most common abnormality of oncogenes in human cancer and a major example of activation by point mutation. Experimental and epidemiological studies support the notion that Ki-ras activation and expression may be chemically related. We discuss the potential role of several environmental compounds in the induction or promotion of ras mutations in humans, with a focus on exocrine pancreatic cancer, the human tumor with the highest prevalence at diagnosis of Ki-ras mutations. Organochlorine compounds, organic solvents, and coffee compounds may play an indirect role in causing Ki-ras mutations, rather than as direct inducers of the mutations. Although for some organochlorine compounds the induction of point mutations in ras oncogenes cannot be excluded, it seems more likely that the effects of these compounds are mediated through nongenomic or indirectly genotoxic mechanisms of action. Organic solvents also may act via enzymatic induction of ras mutagens or by providing a proliferation advantage to ras-mutated cell clones. In exocrine pancreatic cancer, caffeine, other coffee compounds, or other factors with which coffee drinking is associated could modulate Ki-ras activation by interfering with DNA repair, cell-cycle checkpoints, and apoptosis. Asbestos, cigarette smoking, and some dietary factors also may be involved in the initiation or the promotion of Ki-ras mutations in lung and colon cancers. Further development of the mechanistic scenarios proposed here could contribute to a meaningful integration of biological, clinical, and environmental knowledge on the causes of altered ras effects. PMID- 12557260 TI - Molecular determinants of tumor differentiation in papillary serous ovarian carcinoma. AB - In epithelial ovarian cancer, tumor grade is an independent prognosticator whose molecular determinants remain unknown. We investigated patterns of gene expression in well- and poorly differentiated serous papillary ovarian and peritoneal carcinomas with cDNA microarrays. A 6500-feature cDNA microarray was used for comparison of the molecular profiles of eight grade III and four grade I stage III serous papillary adenocarcinomas. With a modified F-test in conjunction with random permutations, 99 genes whose expression was significantly different between grade I and grade III tumors were identified (P < 0.01). A disproportionate number of these differentially expressed genes were located on the chromosomal regions 20q13 and all exhibited higher expression in grade III tumors. Interphase fluorescent in situ hybridization demonstrated 20q13 amplification in two of the four grade III and none of the three grade I tumors available for evaluation. Several centrosome-related genes also showed higher expression in grade III tumors. We propose a model in which tumor differentiation is inversely correlated with the overexpression of several oncogenes located on 20q13, a common amplicon in ovarian and numerous other cancers. Dysregulation of centrosome function is one potential mechanistic link between genetic/epigenetic changes and the poorly differentiated phenotype in ovarian cancer. PMID- 12557262 TI - Exceptional activity of murine glutathione transferase A1-1 against (7R,8S) dihydroxy-(9S,10R)-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene-induced DNA damage in stably transfected cells. AB - We have shown previously that the alpha class murine glutathione transferase (GST) isoenzyme mGSTA1-1, unlike other mammalian class alpha GSTs, is highly efficient in catalyzing the glutathione (GSH) conjugation of (7R,8S)-dihydroxy (9S,10R)-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene [(+)-anti-BPDE], which is the ultimate carcinogenic metabolite of benzo[a]pyrene. The present studies were undertaken to determine the efficacy of mGSTA1-1 in cellular protection against (+)-anti-BPDE-induced DNA damage in HepG2 cells stably transfected with mGSTA1 cDNA. Untransfected HepG2 cells, vector-transfected HepG2 cells (HepG2-vector), and cells transfected with mGSTA4 cDNA (HepG2-mGSTA4), an alpha class murine GST isoenzyme with low (+)-anti-BPDE-GSH conjugating activity, were used as controls for comparison. Intracellular GSH conjugation of (+)-anti-BPDE was significantly higher in mGSTA1-1-overexpressing HepG2 cells (HepG2-mGSTA1) than in HepG2-vector or HepG2-mGSTA4 cells. The formation of DNA-adducts of (+)-anti-BPDE, following a 10-, 20-, or 30-min exposure to 0.1, 0.5, or 1.0 microM [3H](+)-anti-BPDE, was reduced significantly in cells transfected with mGSTA1-1 compared with HepG2 vector or untransfected HepG2 cells. Consistent with the results with purified protein, overexpression of mGSTA4-4 had no effect on (+)-anti-BPDE-induced DNA damage. The results of the present study indicated that mGSTA1-1 was exceptionally effective in affording protection against (+)-anti-BPDE-induced DNA damage in a cellular system. PMID- 12557261 TI - Alterations in the Fhit gene in pancreatic duct adenocarcinomas induced by N nitrosobis(2-oxopropyl)amine in hamsters. AB - Alteration of the Fhit gene was investigated in pancreatic duct adenocarcinomas induced by N-nitrosobis(2-oxopropyl)amine (BOP) in Syrian golden hamsters. The animals received 70 mg/kg BOP, followed by repeated exposure to an augmentation pressure regimen consisting of a choline-deficient diet combined with DL ethionine and then L-methionine and administration of 20 mg/kg BOP. A total of 15 pancreatic duct adenocarcinomas were obtained 10 wk after the beginning of the experiment, and total RNAs were extracted from each for assessment of aberrant transcription of the Fhit gene by reverse transcription-polymerase chain reaction analysis. Aberrant transcripts lacking nucleotides in the regions of nt -75 to 348, nt -15 to 348, or nt -75 to 178 were detected in 11 adenocarcinomas (73.3%). Southern blot analysis of eight tumors did not show any evidence of gross rearrangement or deletion. These results indicated that changes in the Fhit gene occurred frequently and thus may have played a role in the development of pancreatic duct adenocarcinomas induced by BOP in hamsters. PMID- 12557263 TI - The expression of the tumor suppressor gene connexin 26 is not mediated by methylation in human esophageal cancer cells. AB - Gap junctional intercellular communication is thought to play an important role in cell differentiation and tissue homeostasis. Gap junctional intercellular communication is mediated by intercellular channels connecting adjacent cells and composed of connexin (Cx) proteins. Until now, approximately 20 different Cx have been characterized in mammals, and they are expressed in a tissue-specific manner. The downregulation of Cx expression is often observed in tumors and transformed cell lines and is believed to contribute to the loss of proliferating control. Connexin 26 (Cx26) is a Cx constitutively expressed in the normal epithelial esophageal tissue. In the majority of esophageal tumors, Cx26 expression is low or totally absent. CpG island hypermethylation is known to be associated with gene silencing in cancer. Because the promoter and exon 1 region of Cx26 are rich in CpG dinucleotides, we examined whether the loss of Cx26 expression in human esophageal TE cell lines was related to the hypermethylation of this region. We analyzed several TE cell lines derived from different human esophageal carcinomas and exhibiting different levels of Cx26 expression by using methylation-sensitive restriction digestion and Southern blot analysis. We did not find any correlation between the Cx26 expression and the methylation level of the promoter region of the Cx26 gene. Our results suggest that methylation was probably not involved as a primary mechanism of Cx26 regulation in human esophageal cancer cell lines. PMID- 12557264 TI - On the nature of genetic changes required for the development of esophageal cancer. AB - It is clear that genetic mutations are necessary for the development of cancer, but the exact number required is not clear, with estimates ranging from one critical hit (e.g., p53) to dozens or perhaps even hundreds of expression changes (by microarray analysis) or chromosomal aberrations. We have used a mathematical model to estimate the critical number of mutations required for the development of esophageal cancer (EC) and to test for the likelihood of an EC major susceptibility gene. Our results suggest that six or seven mutations are required for the development of EC and that there is no evidence of a major susceptibility gene. This does not exclude the possibility that gene-environment interactions may not confer susceptibility or risk. The gradual accumulation of aberrant gene function also can explain the progression of pathologic states seen in the esophagus, from early dysplasia through mild to severe dysplasia and, finally, to cancer, as illustrated in our model. PMID- 12557265 TI - Identification of three 11p11.2 candidate liver tumor suppressors through analysis of known human genes. AB - We have previously mapped a liver tumor suppressor locus to human chromosome 11p11.2-p12 using a functional model of tumor suppression. Using this model system, we have employed a candidate gene approach to identify potential liver tumor suppressor genes. Thirty-eight known genes have been positioned in human 11p11.2-p12 by the Human Genome Project. Here we show that four of these genes (guanine nucleotide binding protein gamma 3; mitochondrial carrier homolog 2; p53 induced protein (PIG11), and pRDI-BF1-rIZ1 domain containing 11) localized to the minimal liver tumor suppressor region within 11p11.2-p12. In fact, all of these genes mapped to human 11p11.2, allowing refinement of the liver tumor suppressor region to this cytogenetic band. Three of the four genes (mitochondrial carrier homolog 2, PIG11, and pRDI-BF1-rIZ1 domain containing 11) were uniformly expressed by an index panel of suppressed microcell hybrid cell lines, identifying them as candidate liver tumor suppressor genes. In a preliminary analysis of four human hepatocellular carcinoma cell lines (HepG2, Hep3B, SNU398, and SNU449), the transcript for PIG11 was lost or significantly decreased in two of these cell lines (HepG2 and Hep3B), suggesting the potential involvement of PIG11 in some human hepatocellular carcinomas. The results of this study extended our previous knowledge of genes located in the minimal liver tumor suppressor region of human 11p11.2 and identified several candidate liver tumor suppressor genes from this region. Further characterization of these candidates will provide new insight into the role of human 11p11.2 in the molecular pathogenesis of human liver cancer. PMID- 12557266 TI - Electrophysiological and pharmacological characteristics of nigral dopaminergic neurons in the conscious, head-restrained rat. AB - Extracellular single-unit recordings of nigral dopamine (DA) neurons were obtained from conscious rats habituated to having their body suspended in a cloth jacket and their head immobilized in the stereotaxic frame by means of a "restraining platform" permanently fixed to the skull. The electrophysiological characteristics of DA neurons from head-restrained rats and their responses to apomorphine and haloperidol were compared with single-unit recordings obtained from rats lightly and deeply anesthetized with chloral hydrate and from mesencephalic slices. Head-restrained rats showed a higher number of spontaneously active DA neurons and a higher percentage of bursting neurons than lightly and deeply anesthetized rats. Indeed, bursting activity was rare in deeply anesthetized rats and was totally absent in slices. Haloperidol was more potent and effective in stimulating the firing rate and bursting activity in head restrained than in lightly anesthetized rats, while it was virtually ineffective in deeply anesthetized rats and totally ineffective in slices. On the other hand, DA neurons in head-restrained rats showed the same average firing rate as DA neurons in lightly and deeply anesthetized rats and in slices. The potency of apomorphine in inhibiting the firing rate, and that of haloperidol in reversing apomorphine effect, did not vary among the different in vivo preparations. The results suggest that chloral hydrate anesthesia blunts or suppresses not only the excitatory inputs which normally sustain the number of spontaneously active DA neurons and their bursting activity, but also the feedback excitation of DA neurons following haloperidol-induced D(2) receptor blockade. On the other hand, chloral hydrate anesthesia modifies neither D(2) autoreceptor sensitivity to apomorphine and haloperidol nor the automatic genesis of action potentials. The head-restrained rat appears to be an important model for studies into the pharmacology and physiology of DA neurons. PMID- 12557267 TI - Regulation of CRE-mediated transcription in mouse brain by amphetamine. AB - Previous work has demonstrated that acute and chronic administration of amphetamine causes phosphorylation of the transcription factor CREB, the cAMP response element (CRE) binding protein, in striatum, a brain region important for the behavioral actions of the drug. To determine whether such phosphorylation is associated with changes in CREB transcriptional activity, we mapped beta galactosidase (beta-gal) expression in a CRE-LacZ transgenic mouse, in which the beta-gal reporter is downstream of CRE sequences, following acute or chronic amphetamine administration. We found that acute amphetamine induced beta-gal expression in a relatively small number of brain regions, including nucleus accumbens (ventral striatum), amygdala, ventral tegmental area, and locus coeruleus. Chronic amphetamine generally produced greater changes in CRE-mediated transcription in most brain regions and induced CRE-transcription in several regions unaffected by acute drug exposure. Interestingly, amphetamine regulation of CRE activity differed dramatically between males and females. In nucleus accumbens, beta-gal expression colocalized predominantly with dynorphinergic neurons after acute amphetamine administration, while chronic administration induced beta-gal expression in both dynorphinergic and enkephalinergic neurons. In ventral tegmental area, acute and chronic amphetamine induced beta-gal expression mainly in dopaminergic neurons, while induction in the locus coeruleus occurred mainly in nonnoradrenergic neurons. This study identifies several brain regions where CRE-mediated transcription may play a key role in the development of neuronal plasticity associated with amphetamine administration. PMID- 12557268 TI - Noradrenergic modulation of ephedrine-induced hypophagia. AB - The hypophagic action of the sympathomimetic amine ephedrine (EPH) in the rat may reflect actions on central dopaminergic (DA) and noradrenergic (NE) systems. EPH indirectly facilitates DA and NE activity and acts as a partial agonist at alpha(1)-adrenergic receptors. Two approaches were used to assess the possible contribution of NE and DA pathways to EPH-induced hypophagia. In the first, regression analyses of published archival data were computed to characterize the relation between the hypophagic potency values of (-)-(EPH) and related sympathomimetic drugs, including (+)-amphetamine, aminorex, mazindol, and phentermine (data derived from Blosser JC et al., 1987) and the most potent action of these drugs on facilitating NE activity or DA activity in rat brain (data derived from Rothman RB et al., 2001). In the NE analyses, the ED(50) values for these drugs for the inhibition of eating in rats were significantly related (r = 0.91, P = 0.03) to the potency of each drug in facilitating NE activity (either release or inhibition of [(3)H]NE reuptake), whereas in the DA analyses the correlation between ED(50) values and DA activity for these drugs was also significant (r = 0.98, P = 0.003). The regression analyses are thus supportive of a role for NE or DA in the hypophagic capacity of EPH. Although an earlier study noted that administration of the putative DA antagonist pimozide in rats attenuated EPH hypophagia, pimozide exerts similar potency in antagonizing DA receptors and alpha(1)-adrenergic receptors. To clarify the role of alpha(1) adrenoceptors in EPH-induced hypophagia, adult male rats were pretreated with the alpha(1)-adrenergic receptor antagonist prazosin (0.0.5 and 2 mg/kg) prior to the administration of (-)-EPH (0, 5, 10, or 20 mg/kg, IP). Prazosin pretreatment at 2.0 mg/kg significantly attenuated the hypophagia, but not the hypodipsia, induced by administration of 10 mg/kg and by 20 mg/kg (-)-EPH. Collectively, these results confirm a critical contribution of of alpha(1)-adrenoceptors to the hypophagic action of (-)-EPH in rats. PMID- 12557269 TI - Graphical analysis of 2-[18F]FA binding to nicotinic acetylcholine receptors in rhesus monkey brain. AB - External imaging of nicotinic acetylcholine receptors (nAChRs) using techniques such as PET would help to clarify the roles of these receptors in the physiology and pathology of brain function. Here we report the results of quantitative PET studies of cerebral nAChRs with 2-[(18)F]fluoro-A-85380 (2-[(18)F]FA) in rhesus monkeys. Data from dynamic PET scans were analyzed using graphical methods. Binding potential (BP) values of 2.0, 0.4, 0.3, and 0.03 observed in the thalamus (Th), cortex (Cx), striatum (Str), and cerebellum (Cb), respectively, were consistent with the pattern of alpha(4)beta(2) nAChR distribution in monkey brain. The high value of 2-[(18)F]FA-specific binding in the rhesus monkey Th and low level of that in Cb compared with nonspecific accumulation of radioactivity in these structures allowed use of Cb as a reference region for calculation of BP and volume of distribution of specific binding (VDsb) in Th by graphical methods, both with and without the plasma input function. In contrast, estimation of 2 [(18)F]FA specific binding in low-receptor-density regions such as Cx and Str required assessment of nondisplaceable volume of distribution (VDnd) in a separate study and measurement of nonmetabolized radioligand concentrations in the plasma. For accurate quantitation of 2-[(18)F]FA-specific binding by graphical analysis, PET studies should last up to 7 h due to the slow kinetics of 2-[(18)F]FA brain distribution. Further, to avoid substantial underestimation in measured BP values the doses of administered 2-[(18)F]FA should not exceed 0.1 nmol/kg body weight. The findings suggest that 2-[(18)F]FA is a promising ligand for quantitation of nAChRs in human brain. PMID- 12557270 TI - Selegiline potentiates cocaine-induced increases in rodent nucleus accumbens dopamine. AB - Selegiline has been proposed as a treatment for cocaine addiction and studies in humans suggest that it attenuates cocaine's reinforcing effects. Here we assessed the effects of selegiline treatment on cocaine-induced increases in nucleus accumbens (NAc) dopamine (DA) in freely moving rodents. Chronic treatment with selegiline (L-deprenyl, 0.25/mg/kg, 24 days) potentiated cocaine-induced increases in NAc DA from 350-600%. However, this enhanced response was abolished when animals were treated chronically with both cocaine and selegiline. Inasmuch as increases in NAc DA are associated with the reinforcing effects of cocaine, these results obtained in rodents suggest that MAO-A and -B inhibition may not be a suitable strategy to antagonize cocaine's reinforcing effects during cocaine detoxification. On the other hand, chronic selegiline treatment may improve DA deficits, which are thought to contribute to relapse through a decreased response to natural rewards. PMID- 12557271 TI - In vivo muscarinic 2 receptor imaging in cognitively normal young and older volunteers. AB - The precise effects of normal aging on the cholinergic system are unknown, as both in vitro and PET studies have shown conflicting results. In vivo determination of muscarinic receptor distribution and density has been hampered by both poor subtype selectivity and/or blood-brain barrier permeability of known ligands. Previous in vitro and in vivo work with the F-18 labeled muscarinic agonist, 3-(3- (3-[(18)F]Flouropropyl)thio)-1,2,5-thiadiazol-4-yl)-1,2,5,6 tetrahydro-1-methylpyridine ((18)FP-TZTP) suggested the use of (18)FP-TZTP to selectively quantify M2 receptors in humans. In this study, we used (18)FP-TZTP to infer M2 receptor avidity in the brains of 15 healthy younger subjects (mean age = 28.3 +/- 5.5 years) and 20 healthy older subjects (mean age = 62.1 +/- 7.7 years). Corrections for subject motion during the 120-min acquisition and partial voluming (PVC) were performed. A one-tissue compartment model was used to estimate the volumes of distribution (V(T)) of (18)FP-TZTP. Within both groups of subjects, volumes of distribution (K(1)/k(2)) in cortical, subcortical, and cerebellar areas were consistent with M2 receptor topography. Compared to younger subjects older subjects had significantly higher means and standard deviations for the volumes of distribution of (18)FP-TZTP throughout much of the cerebellum, cortex, and subcortex (Global Gray V(T) = 742 +/- 163 in older subjects and 645 +/- 74 in younger subjects, P < 0.03). Across all subjects (18)FP-TZTP, regional, and Global Gray distribution volumes were significantly correlated to age (Global Gray V(T,) r = 0.41, P < 0.01). A lower concentration of acetylcholine in the synapse of some older subjects is one possible explanation for the data. PMID- 12557273 TI - Role of phosphatidylinositol 3-phosphate in formation of forespore membrane in Schizosaccharomyces pombe. AB - Phosphatidylinositol (PI) 3-kinase (encoded by the pik3(+) gene) in Schizosaccharomyces pombe has been identified as a homologue of VPS34p, a protein required for proper vesicular protein sorting. The clone defective in this protein carries enlarged vacuoles and exhibits sensitivity to high temperature or high ion concentration. The effect of disruption of pik3(+) on sporulation of Sz. pombe was examined. The diploid cells underwent G(1) arrest and meiosis. However, the spores formed by the deltapik3 cells were not viable. Electron-microscopic analysis revealed that the growth of the forespore membrane of deltapik3 cells was not correctly orientated, failing to engulf the nucleus or forming extremely small spores, as was confirmed by the use of Spo3p-GFP and GFP-Psy1p, which are markers of the forespore membrane. The coating materials found along the forespore membrane of the wild-type were greatly reduced in these cells. PI 3-P, the product of Pik3p, was detected on the forespore membrane, suggesting that PI 3-P-dependent vesicle transport may take place in formation of the forespore membrane. Misshaped forespore membrane, accumulation of vesicles, formation of small non-viable spores, and suppression by over expression of Psy1p were the phenotypes commonly seen in deltapik3 and deltaspo3 cells, suggesting a relationship between the functions of Pik3p and Spo3p in formation of the forespore membrane in Sz. pombe. PMID- 12557272 TI - D1 dopamine receptor regulates alcohol-motivated behaviors in the bed nucleus of the stria terminalis in alcohol-preferring (P) rats. AB - Recent studies have implicated the bed nucleus of the stria terminalis (BST) as a potential brain substrate for mediating drug-related behaviors. Neuroanatomical studies have demonstrated that reciprocal projections exist from the BST to the ventral tegmental area (VTA), a dopamine reward substrate proposed to play a role in alcohol abuse. In the present study, we evaluated the role of the D(1) and D(2) dopamine receptors of the BST in regulating alcohol and sucrose-motivated behaviors. Alcohol-preferring (P) rats were trained under an FR4 operant schedule to self-administer either EtOH (10% v/v) or sucrose (2% w/v). Following training, we evaluated the capacity of a competitive D(1) (SCH 23390; 0.5-20.0 microg) and a D(2) (eticlopride; 0.5-20.0 microg) dopamine antagonist to selectively reduce EtOH-maintained responding. Naltrexone, (5-30.0 microg), the nonselective opioid antagonist, was used as a reference agent. The results showed that SCH 23390 dose dependently reduced alcohol-motivated responding. Responding was reduced with the 20.0 microg dose to about 97% of control levels. SCH 23390 also reduced sucrose responding; however, the magnitude of effects was substantially lower with the highest doses (2.5, 20.0 microg) (68-79% of control levels). In contrast, eticlopride failed to significantly alter alcohol responding and reduced sucrose responding only with the 10.0 microg dose. Unlike the dopamine antagonists, all naltrexone doses failed to alter EtOH or sucrose-maintained responding. The results suggest a salient role for the D(1), but not the D(2) and opioid receptors in selectively modulating EtOH-motivated behaviors in the BST. PMID- 12557274 TI - Transcription patterns of PMA1 and PMA2 genes and activity of plasma membrane H+ ATPase in Saccharomyces cerevisiae during diauxic growth and stationary phase. AB - PMA1 and PMA2 genes encode Saccharomyces cerevisiae plasma membrane H(+)-ATPase (PM-H(+)-ATPase), an enzyme with critical physiological roles both in the absence or presence of environmental stress. The two PM-H(+)-ATPase isoforms differ in their biochemical characteristics but, under all the growth conditions that were examined so far, PMA2 expression is negligible and Pma1p accounts for practically the totality of cell PM-H(+)-ATPase activity. In the present work, we have compared gene expression levels and activity of this proton pump in yeast cells cultivated under fermentative or respiratory growth and under carbon starvation. The expression levels of both PMA1 and PMA2 genes were consistently higher (2.5 4.5-fold) in cells cultivated under respiratory metabolism (in ethanol-based medium or after the diauxic shift), than in cells cultivated under fermentative metabolism (during the full period of growth in a medium where glucose is not the limiting nutrient or during the first period of diauxic growth in low-glucose based medium). The moderate upregulation of PMA1 and PMA2 transcription in cells grown on ethanol compared with those grown on glucose was reflected in the increased content and activity of PM-H(+)-ATPase. In diauxic growth, during transition to stationary phase after ethanol depletion, a further strong activation (eight-fold) of PMA2 gene transcription was observed. Although PMA2 transcription still remains quite below (20-fold) PMA1 transcription, this is the first environmental condition, identified so far, that leads to a significant PMA2 expression, suggesting that this PM-H(+)-ATPase isoform may play some role during carbon starvation. PMID- 12557275 TI - Isolation and characterization of the plasma membrane biotin transporter from Schizosaccharomyces pombe. AB - The fission yeast Schizosaccharomyces pombe is auxotrophic for biotin (vitamin H) and growth depends on biotin uptake over the plasma membrane. Here a biotin transport mutant of Saccharomyces cerevisiae is used to identify the vht1(+) gene encoding the Schizosaccharomyces pombe plasma membrane transport protein for biotin. SpVht1p belongs to the family of allantoate transporters and has only little sequence homology to the S. cerevisiae biotin transporter. Although having dissimilar primary structures, the biotin transporters in Sz. pombe and S. cerevisiae share similar biochemical properties and regulation. Like in S. cerevisiae, biotin uptake in Sz. pombe is a high-affinity process, is optimal at acidic pH values and inhibited by protonophores, indicating that SpVht1p acts as a proton-biotin symporter. Desthiobiotin, the metabolic precursor of biotin, is also imported by SpVht1p. Deletion of vht1(+) abolishes growth on low external concentrations of the vitamin, showing that vht1(+) encodes the only protein that mediates biotin uptake in Sz. pombe. Expression of vht1(+) is maximal at low external biotin concentrations, indicating that Sz. pombe can adjust the rate of biotin uptake to meet the requirement for the vitamin. PMID- 12557276 TI - Identification of a triacylglycerol lipase gene family in Candida deformans: molecular cloning and functional expression. AB - The yeast Candida deformans CBS 2071 produces an extracellular lipase which was shown to catalyse the production of various esters by the esterification of free fatty acids, even in the presence of a large molar excess of water. To clone the gene encoding this extracellular lipase, Saccharomyces cerevisiae was transformed with C. deformans genomic libraries and screened for lipolytic activity on a medium containing rapeseed oil emulsion and rhodamine B. Three members of a lipase gene family (CdLIP1, CdLIP2 and CdLIP3) were cloned and characterized. Each deduced lipase sequence has a Gly-His-Ser-Leu-Gly-(Gly/Ala)-Ala conserved motif, eight cysteine residues and encodes an N-terminal signal sequence. MALDI TOF mass spectrometry analysis of a proteolytic digest of the lipase produced was used to obtain experimental evidence that the CdLIP1 gene encoded the extracellular lipase. Recombinant expression studies confirmed that the cloned genes encoded functional lipases. The three lipases are very similar to lipases from the related species Yarrowia lipolytica. Significant homologies were also found with several yeast and fungal lipases. As C. deformans CBS 2071 was previously considered to be synonymous with Y. lipolytica, the strains were compared for the extent of nucleotide divergence in the variable regions (D1/D2) at the 5'-end of the large-subunit (26S) ribosomal DNA (rDNA) gene. This rDNA region has diverged sufficiently to suggest that C. deformans is a separate species. The nucleotide sequences of the CdLIP1, CdLIP2 and CdLIP3 genes will appear in the EMBL nucleotide sequence database under Accession Nos AJ428393, AJ428394 and AJ428395, respectively. PMID- 12557277 TI - Function of Candida glabrata ABC transporter gene, PDH1. AB - The rapid increase in azole resistance during treatment of patients infected with Candida glabrata may be due to increased azole efflux mediated by ABC transporters, as occurs with increased expression of PDR5 in Saccharomyces cerevisiae. Two known C. glabrata homologues of PDR5 influencing azole susceptibility are PDH1 (CgCDR2) and CgCDR1. Disruption of PDH1 in a cgcdr1::ura3 strain increased susceptibility to rhodamine 6G, cycloheximide and chloramphenicol, and also increased rhodamine 6G accumulation, all properties of pdr5 null mutants. Overexpression of PDH1 in S. cerevisiae complemented the pdr5 mutation by reversing susceptibility to rhodamine 6G, chloramphenicol and cycloheximide, as well as by decreasing rhodamine 6G intracellular concentration. Expression of PDH1 in a C. glabrata cgcdr1::ura3 pdh1Delta::ura3 mutant using a multicopy plasmid almost completely restored the wild-type phenotype, showing that PDH1 at higher levels of expression can replace CgCDR1. Because PDH1 and CgCDR1 have both been reported to have upstream sequences similar to the Pdr1p- and Pdr3p-binding elements of PDR5, we sought similarities in regulation between the three genes. Abundance of PDH1 and CgCDR1 mRNA in C. glabrata was increased by rhodamine 6G, cycloheximide and oligomycin, properties in common with PDR5. PDH1, CgCDR1 and PDR5 have striking similarities in function and regulation. PMID- 12557279 TI - Risk factors for childhood stroke: little folks have different strokes! PMID- 12557280 TI - Nitric oxide and the axonal death cascade. PMID- 12557281 TI - The neurobiology, diagnosis, and treatment of narcolepsy. AB - Narcolepsy is a common cause of chronic sleepiness distinguished by intrusions into wakefulness of physiological aspects of rapid eye movement sleep such as cataplexy and hallucinations. Recent advances provide compelling evidence that narcolepsy may be a neurodegenerative or autoimmune disorder resulting in a loss of hypothalamic neurons containing the neuropeptide orexin (also known as hypocretin). Because orexin promotes wakefulness and inhibits rapid eye movement sleep, its absence may permit inappropriate transitions between wakefulness and sleep. These discoveries have considerably improved our understanding of the neurobiology of sleep and should foster the development of rational treatments for a variety of sleep disorders. PMID- 12557282 TI - Investigation of risk factors in children with arterial ischemic stroke. AB - We present data on the known risk factors encountered in children presenting with a first arterial ischemic stroke to a single tertiary center over 22 years. Two hundred twelve patients (54% male; median age, 5 years) were identified. One hundred fifteen (54%) were previously healthy. Cerebral arterial imaging was undertaken in 185 patients (87%) and was abnormal in 79%. Of 104 previously healthy patients investigated with echocardiography, only 8 had abnormal studies. Genetic or acquired conditions causing thrombophilia were rare. Forty percent of patients were anemic, and 21% either had elevated total plasma homocysteine or were homozygous for the t-MTHFR mutation. Trauma and previous varicella zoster infection were significantly more common in the previously healthy group. There was a significant association between cerebral arterial abnormalities and systolic blood pressure greater than 90th percentile and a trend for an association with varicella within the previous year. Clinical history and examination usually identify underlying risk factors and precipitating triggers for arterial ischemic stroke in childhood. Cerebral arterial imaging is usually abnormal, but echocardiography and prothrombotic screening are commonly negative. PMID- 12557283 TI - Blockers of sodium and calcium entry protect axons from nitric oxide-mediated degeneration. AB - Axonal degeneration can be an important cause of permanent disability in neurological disorders in which inflammation is prominent, including multiple sclerosis and Guillain-Barre syndrome. The mechanisms responsible for the degeneration remain unclear, but it is likely that axons succumb to factors produced at the site of inflammation, such as nitric oxide (NO). We previously have shown that axons exposed to NO in vivo can undergo degeneration, especially if the axons are electrically active during NO exposure. The axons may degenerate because NO can inhibit mitochondrial respiration, leading to intraaxonal accumulation of Na(+) and Ca(2+) ions. Here, we show that axons can be protected from NO-mediated damage using low concentrations of Na(+) channel blockers, or an inhibitor of Na(+)/Ca(2+) exchange. Our findings suggest a new strategy for axonal protection in an inflammatory environment, which may be effective in preventing the accumulation of permanent disability in patients with neuroinflammatory disorders. PMID- 12557284 TI - Assessing the onset of structural change in familial Alzheimer's disease. AB - Regional and global cerebral atrophy are inevitable features of Alzheimer's disease (AD). We assessed volumes and atrophy rates of brain structures in patients with familial AD during the period that they developed symptoms. Five patients with presymptomatic AD and 20 controls had two or more annual volumetric MRI brain scans. Volumes of brain, ventricles, temporal lobes, hippocampi, and entorhinal cortices (ECs) were measured. Rates of volume change were calculated from serial scans. There were no significant differences in baseline measures of whole brain, temporal lobe, or ventricular volume between patients and controls; averaged volumes of medial temporal lobe structures (both hippocampi and ECs) were 16.6% (95% confidence interval [CI], 3.3-28.0%) lower in patients. Atrophy rates for brain, temporal lobe, hippocampus, and EC were significantly increased in patients compared with controls (p < 0.05). Averaged atrophy rates from both hippocampi and ECs were 5.1% (95% CI, 3.0-7.1%) greater in patients than controls. Linear extrapolation backward suggested medial temporal lobe atrophy commenced 3.5 years (95% CI, 0.7-7.5 years) before onset, when all patients were asymptomatic. We conclude that increased medial temporal lobe atrophy rates are an early and distinguishing feature of AD and that pathological atrophy probably is occurring several years before the onset of symptoms. PMID- 12557285 TI - Cross-reactivity with myelin basic protein and human herpesvirus-6 in multiple sclerosis. AB - Viral infections are though to play an important role in the pathogenesis of multiple sclerosis (MS) potentially through molecular mimicry. An identical sequence was found in both myelin basic protein (MBP, residues 96-102), a candidate autoantigen for MS, and human herpesvirus-6 (HHV-6 U24, residues 4-10) that is a suspected viral agent associated with MS. In this study, we showed that greater than 50% of T cells recognizing MBP(93-105) cross-reacted with and could be activated by a synthetic peptide corresponding to residues 1 to 13 of HHV-6 U24 in MS patients. The estimated precursor frequency of these cross-reactive T cells recognizing both peptides, MBP(93-105) and HHV-6 (U24)(1-13), was significantly elevated in MS patients compared with that in healthy controls. These cross-reactive CD4+ T cells represented the same Th1 phenotype as that of monospecific T cells recognizing MBP(93-105). There were increased antibody titers for both peptide HHV-6 (U24)(1-13) and peptide MBP(93-105) in the same patients with MS compared with those in healthy controls, suggesting B-cell sensitization to the antigens in MS patients. The study provides important evidence in the understanding of the potential role of HHV-6 infection/reactivation in the activation of autoimmune reactivity to MBP and its implication in the pathogenesis of MS. PMID- 12557286 TI - Suppression of complex I gene expression induces optic neuropathy. AB - Optic nerve degeneration is a feature common to diseases with mutations in genes that encode complex I of the respiratory chain. Vulnerability of this central nervous system tract is a mystery, because of the paucity of animal models used to investigate effects of the mutated DNA in tissues rather than isolated in cultured cells. Using a ribozyme designed to degrade the mRNA encoding a critical nuclear-encoded subunit gene of complex I (NDUFA1), we tested whether oxidative phosphorylation deficiency can recapitulate the optic neuropathy of mitochondrial disease. Injection of adenoassociated virus expressing this ribozyme led to axonal destruction and demyelination, the hallmarks of Leber hereditary optic neuropathy. PMID- 12557278 TI - Transcriptome characterization of the dimorphic and pathogenic fungus Paracoccidioides brasiliensis by EST analysis. AB - Paracoccidioides brasiliensis is a pathogenic fungus that undergoes a temperature dependent cell morphology change from mycelium (22 degrees C) to yeast (36 degrees C). It is assumed that this morphological transition correlates with the infection of the human host. Our goal was to identify genes expressed in the mycelium (M) and yeast (Y) forms by EST sequencing in order to generate a partial map of the fungus transcriptome. Individual EST sequences were clustered by the CAP3 program and annotated using Blastx similarity analysis and InterPro Scan. Three different databases, GenBank nr, COG (clusters of orthologous groups) and GO (gene ontology) were used for annotation. A total of 3,938 (Y = 1,654 and M = 2,274) ESTs were sequenced and clustered into 597 contigs and 1,563 singlets, making up a total of 2,160 genes, which possibly represent one-quarter of the complete gene repertoire in P. brasiliensis. From this total, 1,040 were successfully annotated and 894 could be classified in 18 functional COG categories as follows: cellular metabolism (44%); information storage and processing (25%); cellular processes-cell division, posttranslational modifications, among others (19%); and genes of unknown functions (12%). Computer analysis enabled us to identify some genes potentially involved in the dimorphic transition and drug resistance. Furthermore, computer subtraction analysis revealed several genes possibly expressed in stage-specific forms of P. brasiliensis. Further analysis of these genes may provide new insights into the pathology and differentiation of P. brasiliensis. PMID- 12557287 TI - Plasticity of the nigropallidal pathway in Parkinson's disease. AB - The degeneration of nigrostriatal dopamine neurons in early Parkinson's disease (PD) is compensated in part by increased transmitter turnover in surviving neurons of the pathway. In this (18)F-dopa positron emission tomography study, we demonstrate compensatory changes in PD in another midbrain dopamine projection to the basal ganglia, the nigropallidal projection to the internal segment of the globus pallidus (GPi). Increased (18)F-dopa uptake in the GPi is seen in early PD which then is lost in advanced PD. Our early PD cases show an absence of significant clinical progression in the face of a continuing loss of nigrostriatal projections. This indicates a compensatory neuronal plasticity that we now show to involve the nigropallidal dopamine pathway to the GPi but not to the external segment of the globus pallidus (GPe). Enhanced function of the dopamine projection to the GPi serves, we propose, to maintain a more normal pattern of pallidal output to ventral thalamus and motor cortex in early PD, whereas loss of this adaptive pathway in advanced disease may be a pivotal step in the progression of the disease. PMID- 12557288 TI - Homocysteine, white matter hyperintensities, and cognition in healthy elderly people. AB - Hyperhomocysteinemia is associated with an increased risk of vascular disease, and recent results suggest that it also could increase the risk of dementia. We examined the relationship between homocysteine and cognitive decline in 1,241 subjects aged 61 to 73 years, followed up over 4 years. Plasma homocysteine levels were determined in all participants as well as cardiovascular risk factors, apolipoprotein E genotype, plasma levels of folate, and vitamin B12. Cognitive performances were assessed repeatedly by using Mini-Mental State Examination, Trail Making Test, Digit Symbol Substitution Test, and Finger Tapping Test. At 2-year follow-up, 841 subjects underwent cerebral magnetic resonance imaging, and white matter hyperintensities were rated visually. Analyses were adjusted for all cardiovascular risk factors. Cross-sectional analyses showed that higher concentrations of homocysteine were significantly related to poorer performances at all neuropsychological tests. Longitudinal analyses confirmed this finding. The odds of cognitive decline was 2.8-fold (p < 0.05) higher in subjects with homocysteine levels above 15 micromol/L compared with those with homocysteine levels below 10 micromol/L. In participants who underwent magnetic resonance imaging, the relationship between homocysteine and cognition was unchanged after taking into account white matter hyperintensities suggesting that white matter hyperintensities do not mediate the association between homocysteine and cognition. PMID- 12557289 TI - Intrathecal IgM synthesis is a prognostic factor in multiple sclerosis. AB - Intrathecal IgM synthesis (ITMS) predicts a worse evolution in the first stages of multiple sclerosis (MS). The aim of this study was the follow-up of a group of relapsing-remitting MS patients for a longer time to evaluate whether the ITMS implies a poor prognosis. Oligoclonal IgM bands were performed in 29 MS patients followed up from 5 to 16 years. Time to conversion to secondary-progressive MS (SPMS), time elapsed to reach a disability of 6 in the Expanded Disability Status Scale (EDSS), percentage of patients with a benign MS, and changes in EDSS score were evaluated. During the follow-up, 70.8% of patients with ITMS converted to SPMS. None of the patients without ITMS did. At the end of the study, 63.6% of patients with ITMS had reached EDSS 6, whereas none of the patients lacking ITMS reached values above EDSS 3. When patients with benign MS were analyzed, 82% lacked ITMS. All patients with a nonbenign MS had ITMS. At the end of the study, the mean EDSS score was 4.64 in patients with ITMS and 1.31 in those without. The presence of oligoclonal IgM bands in cerebrospinal fluid is an unfavorable prognostic marker in MS. PMID- 12557290 TI - Magnetic resonance cerebral metabolic rate of oxygen utilization in hyperacute stroke patients. AB - The purpose of this study was to explore the feasibility of obtaining magnetic resonance-measured cerebral metabolic rate of oxygen utilization (MR-CMRO(2)) in acute ischemic stroke patients. Seven stroke patients were serially imaged: 4.5 +/- 0.9 hours (tp1), 3 to 5 days (tp2), and 1 to 3 months (tp3) after symptom onset. Diffusion-weighted, perfusion-weighted, and multiecho gradient-echo/spin echo images were acquired; cerebral blood flow and oxygen extraction fraction maps were obtained from which CMRO(2) was calculated as the product of cerebral blood flow and oxygen extraction fraction. The final infarct lesions obtained from tp3 T2-weighted images and the "penumbra" obtained from the tp1 perfusion weighted image-defined lesion were coregistered onto tp1 CMRO(2) maps. CMRO(2) values in the region of brain that eventually infarcted were reduced to 0.40 +/- 0.24 of the respective region on the contralateral hemisphere. The "salvaged penumbra" defined by the area of mismatch between the final infarct and the tp1 perfusion-weighted lesion demonstrated an average CMRO(2) value of 0.55 +/- 0.11 of the contralateral hemisphere. Although our results are preliminary and require further evaluation, the ability to obtain in vivo measurements of MR-CMRO(2) noninvasively potentially can provide information for determining brain tissue viability in acute ischemic stroke patients. PMID- 12557291 TI - Consequences of a novel caveolin-3 mutation in a large German family. AB - Mutations in the human caveolin-3 gene (cav-3) on chromosome 3p25 have been described in limb girdle muscular dystrophy, rippling muscle disease, hyperCKemia, and distal myopathy. Here, we describe the genetic, myopathological, and clinical findings in a large German family harboring a novel heterozygous mutation (GAC-->GAA) in codon 27 of the cav-3 gene. This missense mutation causes an amino acid change from asparagine to glutamate (Asp27Glu) in the N-terminal region of the Cav-3 protein, which leads to a drastic decrease of Cav-3 protein expression in skeletal muscle tissue. In keeping with an autosomal dominant mode of inheritance, this novel cav-3 mutation was found to cosegregate with neuromuscular involvement in the reported family. Ultrastructural analysis of Cav 3-deficient muscle showed an abnormal folding of the plasma membrane as well as multiple vesicular structures in the subsarcolemmal region. Neurological examination of all nine subjects from three generations harboring the novel cav-3 mutation showed clear evidence of rippling muscle disease. However, only two of these nine patients showed isolated signs of rippling muscle disease without muscle weakness or atrophy, whereas five had additional signs of a distal myopathy and two fulfilled the diagnostic criteria of a coexisting limb girdle muscular dystrophy. These findings indicate that mutations in the human cav-3 gene can lead to different and overlapping clinical phenotypes even within the same family. Different clinical phenotypes in caveolinopathies may be attributed to so far unidentified modifying factors/genes in the individual genetic background of affected patients. PMID- 12557292 TI - Axonal injury within language network in primary progressive aphasia. AB - Primary progressive aphasia (PPA) is characterized by an isolated progressive impairment of word use and comprehension reflecting the distribution of pathological processes within the left hemisphere. We used proton magnetic resonance spectroscopy (1H-MRS) to study in vivo the integrity of axonal fibers connecting perisylvian language areas in 11 patients with PPA, 11 subjects with Alzheimer's disease, and 22 controls. Brain metabolites (N-acetylaspartate, myoinositol, choline, creatine) were measured bilaterally within a volume of interest located in the central portion of the superior longitudinal fasciculus, a long associative bundle connecting Broca's area with Wernicke's area, and other language regions of the temporal lobe. In the PPA group, there was an asymmetrical N-acetylaspartate to creatine ratio reduction compared with Alzheimer's disease and controls, with greater changes on the left side. The myoinositol to creatine ratio was increased in the PPA group bilaterally compared with controls. The choline to creatine ratio did not differ among the three groups. These results indicate an asymmetrical focal axonal injury within the language network in PPA. The marked difference in the distribution of N acetylaspartate to creatine between PPA and Alzheimer's disease suggests that proton magnetic resonance spectroscopy may help to differentiate between these two conditions. PMID- 12557293 TI - Lack of creatine in muscle and brain in an adult with GAMT deficiency. AB - Guanidinoacetate methyltransferase deficiency, which so far has been exclusively detected in children, was diagnosed in a 26-year-old man. The full-blown spectrum of clinical symptoms already had been present since infancy without progression of symptoms during adolescence. Cranial magnetic resonance imaging showed normal findings. Ophthalmological examination showed no retinal changes. Besides creatine deficiency in the brain, a distinct lack of phosphocreatine in skeletal muscle was proved by (31)P magnetic resonance spectroscopy. Creatine substitution combined with a guanidinoacetate-lowering diet introduced first at the age of 26 years was shown to be effective by an impressive improvement of epileptic seizures, mental capabilities, and general behavior and by normalization of the (31)P spectrum in the skeletal muscle. PMID- 12557294 TI - A new leukoencephalopathy with brainstem and spinal cord involvement and high lactate. AB - We identified eight patients with a distinct magnetic resonance imaging pattern of inhomogeneous cerebral white matter abnormalities and selective involvement of brainstem and spinal tracts. Proton magnetic resonance imaging showed increased lactate in the abnormal white matter. Clinically, the patients had slowly progressive pyramidal, cerebellar, and dorsal column dysfunction. The uniform, highly characteristic magnetic resonance imaging pattern and the similarities in clinical and magnetic resonance spectroscopy findings provide evidence for a new disease entity. Autosomal recessive inheritance is likely. PMID- 12557295 TI - Magnetic resonance imaging and neurosphere therapy of stroke in rat. AB - We intracisternally transplanted subventricular zone (SVZ) cells labeled by ferromagnetic particles into stroked rats. Migration of transplanted cells was non-invasively tracked using magnetic resonance imaging (MRI). We found that transplanted cells selectively migrated towards the ischemic parenchyma at a mean speed of 65 +/- 14.6 microm/hr in living rats. Migration of transplanted cells in the brain was also measured histopathologically. Rats transplanted with SVZ cells exhibited significant improvement of neurological function. Our data suggest that intracisternal transplantation of SVZ cells provides an avenue for cell therapy of stroke and that MRI can be used to track grafted cells in the brain. PMID- 12557296 TI - Transient suppression of tinnitus by transcranial magnetic stimulation. AB - It has been proposed that tinnitus is associated with an irregular activation of the temporoparietal cortex. If this activity is a functionally relevant component of the tinnitus-related network, a virtual temporary lesion of this area should result in transient reduction of tinnitus. To test this hypothesis, we applied 10Hz repetitive transcranial magnetic stimulation to eight scalp and four control positions in 14 patients with chronic tinnitus. Stimulation of left temporoparietal cortex significantly reduced tinnitus (Friedman analysis of variance, p < 0.05; compared with control), indicating that secondary auditory areas can be critical for tinnitus perception, perhaps as a consequence of maladaptive cortical reorganization. PMID- 12557297 TI - Additive neuroprotective effects of minocycline with creatine in a mouse model of ALS. AB - The known neuroprotective effects of minocycline and creatine in animal models of amyotrophic lateral sclerosis (ALS) led us to examine whether the combination of these agents would result in increased neuroprotection. As previously reported, we confirmed in ALS mice that either minocycline or creatine treatment results in improvement in motor performance and extended survival. We report that combination of minocycline and creatine resulted in additive neuroprotection, suggesting this to be a novel potential strategy for the treatment of ALS. To our knowledge, this is the first report demonstrating additive neuroprotection of a combinatorial approach in a mouse model of ALS. Adding relevancy to our findings, minocycline and creatine, are relatively safe, cross the blood-brain barrier, and are currently available for human evaluation. PMID- 12557298 TI - Oculobulbar involvement is typical with Lambert-Eaton myasthenic syndrome. AB - Oculobulbar symptoms and/or signs were present in 18 of 23 (78%) of Lambert-Eaton myasthenic syndrome (LEMS) patients evaluated at the Lahey Clinic (Table). Sixty five percent (15 of 23) of our patients had ptosis and/or diplopia, each present in 11 individuals. Bulbar signs and symptoms, including dysarthria in 10 and dysphagia in 8 patients, also were observed among our LEMS population. More than one prereferral oculobulbar feature occurred in 13 of our LEMS patients. Prereferral diagnostic considerations included myasthenia gravis, myopathies, and psychiatric disorders. These findings suggest that these atypical characteristics served to dissuade some colleagues from a diagnosis of LEMS. Thus, the presence of oculobulbar symptoms and signs cannot be used to exclude LEMS from the differential diagnosis. PMID- 12557299 TI - A new case of pyruvate dehydrogenase deficiency due to a novel mutation in the PDX1 gene. AB - We report a case of neonatal congenital lactic acidosis associated with pyruvate dehydrogenase E3-binding protein deficiency in a newborn girl. She had a severe encephalopathy, and magnetic resonance imaging of the brain showed large subependymal cysts and no basal ganglia lesions. She died 35 days after birth. We detected a novel homozygous deletion (620delC) in the PDX1 gene, which encodes for the E3BP subunit of the pyruvate dehydrogenase complex. PMID- 12557300 TI - The V368i mutation in Twinkle does not segregate with AdPEO. PMID- 12557301 TI - The promoter region of the Menkes gene ATP7A is not altered in focal or generalized dystonia. PMID- 12557303 TI - Prodynorphin gene promoter polymorphism and temporal lobe epilepsy. PMID- 12557304 TI - Human herpesvirus-6 encephalitis associated with hypersensitivity syndrome. PMID- 12557308 TI - Combined transcriptome and proteome analysis of Escherichia coli during high cell density culture. AB - Combined transcriptome and proteome analysis was carried out to understand metabolic and physiological changes of Escherichia coli during the high cell density cultivation (HCDC). The expression of genes of TCA cycle enzymes, NADH dehydrogenase and ATPase, was up-regulated during the exponential fed-batch period and was down-regulated afterward. However, expression of most of the genes involved in glycolysis and pentose phosphate pathway was up-regulated at the stationary phase. The expression of most of amino acid biosynthesis genes was down-regulated as cell density increased, which seems to be the major reason for the reduced specific productivity of recombinant proteins during HCDC. The expression of chaperone genes increased with cell density, suggesting that the high cell density condition itself can be stressful to the cells. Severe competition for oxygen at high cell density seemed to make cells use cytochrome bd, which is less efficient but has a high oxygen affinity than cytochrome bo(3). Population cell density itself strongly affected the expression of porin protein genes, especially ompF, and hence the permeability of the outer membrane. Expression of phosphate starvation genes was most strongly up-regulated toward the end of cultivation. It was also found that sigma(E) (rpoE) plays a more important role than sigma(S) (rpoS) at the stationary phase of HCDC. These findings should be invaluable in designing metabolic engineering and fermentation strategies for the production of recombinant proteins and metabolites by HCDC of E. coli. PMID- 12557309 TI - An EPR study of the secretion of G-CSF heterologous protein from Pichia pastoris. AB - The biologically active protein known as human granulocyte colony stimulating factor (G-CSF) can be efficiently secreted from the transformed GS115 Pichia pastoris (GS115/pPIC9/G-CSF), which contains an alpha-mating-factor prepro signal sequence and an alcohol oxidase I promoter, both introduced using the pPIC9 plasmid. To study the P. pastoris G-CSF protein-secretion process, changes to the plasma membrane's lateral domain structure were monitored using electron paramagnetic resonance (EPR). The GS115 and its transformed analogue show that the plasma membrane can be described by fluid-disordered and fluid-ordered lateral domains. The relative proportion of these domains is a sensitive parameter that is able to describe the membrane's involvement in the protein excretion process. Here, P. pastoris GS115 served as a control for us to compare with the GS115/pPIC9/G-CSF heterologous protein-secreting cells. Electron paramagnetic resonance studies using the spin-probe 5-doxyl methyl ester of palmitic acid [MeFASL (10,3)] showed that during cultivation in a glycerol medium all types of cells had a relatively high proportion of cell-membrane fluid disordered domains, while during the production phase the G-CSF protein-secreting cells showed a decrease in the proportion of fluid-disordered domains. We ascribe this effect to the vesicular lipid exchange caused by the fusion of secretary vesicles with the plasma membrane during exocytosis. Using electron microscopy and immunocytochemistry intracellular vesicles containing the G-CSF protein were detected. Our studies support the exocytotic mechanism of the heterologous protein secretion. PMID- 12557310 TI - Bioprocess considerations for expanded-bed chromatography of crude canola extract: sample preparation and adsorbent reuse. AB - Compared to the conventional microbial and mammalian systems, transgenic plants produce proteins in a different matrix. This provides opportunities and challenges for downstream processing. In the context of the plant host Brassica napus (canola), this work addresses the bioprocessing challenges of solid fractionation, resin fouling by native plant components (e.g., oil, phenolics, etc.), hydrodynamic stability, and resin reuse for expanded bed adsorption for product capture. Plant tissue processing and subsequent protein extraction typically result in an extract with a high content of solids containing a wide particle-size distribution. Without removal of larger particles, the column inlet distributor plugged. The larger particles (> 50 microm) were easily removed through centrifugal settling comparable to that attainable with a scroll decanter. The remaining solids did not affect the column performance. Less than 4% of the lipids and phenolics in the fed extract bound to STREAMLINE trade mark DEAE resin, and this small proportion could be satisfactorily removed using recommended clean-in-place (CIP) procedures. Hydrodynamic expansion and adsorption kinetics of the STREAMLINE trade mark DEAE resin were maintained throughout 10 cycles of reuse, as was the structural integrity of the resin beads. No significant accumulation of N-rich (e.g., proteins) and C/O-rich components (e.g., oil and phenolics) occurred over the same period. PMID- 12557311 TI - Simultaneous synthesis of enantiomerically pure (S)-amino acids and (R)-amines using coupled transaminase reactions. AB - For the simultaneous synthesis of enatiomerically pure (S)-amino acids and (R) amines from corresponding alpha-keto acids and racemic amines, an alpha/omega transaminase coupled reaction system was designed using favorable reaction equilibrium shift led by omega-transaminase reaction. Cloned tyrB, aspC and avtA, and omegataA were co-expressed in E. coli BL21(DE3) using pET23b(+) and pET24ma, respectively. The coupled reaction produced the (S)-amino acids with 73-90% (> 99% ee(S)) of conversion yield and resolved the racemic amines with 83-99% ee(R) for 5 to 10 hours. In designing the coupled reactions in the cell, alanine and pyruvate were efficiently used in the cell as an amine donor for the alanine transaminase and an amino acceptor for the omega-transaminase, respectively, resulting in an alanine-pyruvate shuttling system. The common problem of the low equilibrium constant of the alpha-transaminase can be efficiently overcome by the coupling with the omega-transaminase. However, overcoming the product inhibition of omega-transaminase by the ketone by-product and increasing the decarboxylation rate of the oxaloacetate produced during the transaminase reaction become barriers to further improving the overall reaction rate and the yield of the coupled reactions. PMID- 12557312 TI - Modeling of scale-down effects on the hydrodynamics of expanded bed adsorption columns. AB - Expanded-bed adsorption (EBA) is a technique for primary recovery of proteins starting from unclarified broths. This process combines centrifugation, concentration, filtration, and initial capturing of the proteins in a single step. An expanded bed (EB) is comparable to a packed bed in terms of separation performance but its hydrodynamics are that of a fluidized bed. Downstream process development involving EBA is normally carried out in small columns to minimize time and costs. Our purpose here is to characterize the hydrodynamics of expanded beds of different diameters, to develop scaling parameters that can be reliably used to predict separation efficiency of larger EBA columns. A hydrodynamic model has been developed which takes into account the radial liquid velocity profile in the column. The scale-down effect can be characterized in terms of apparent axial dispersion, D(axl,app), and plate number, N(EB), adapted for expanded bed. The model is in good agreement with experimental results obtained from 1- and 5-cm column diameters with buffer solutions of different viscosities. The model and the experiments show an increase of apparent axial dispersion with an increase in column diameter. Furthermore, the apparent axial dispersion is affected by an increase in liquid velocity and viscosity. Supported by visual observations and predictions from the model, it was concluded that operating conditions (liquid viscosity and superficial velocity) resulting in a bed-void fraction between 0.7 and 0.75 would provide the optimal separation efficiency in terms of N(EB). PMID- 12557313 TI - Surface engineering of living myoblasts via selective periodate oxidation. AB - Cell surface molecules are vital for normal cell activity. To study the functions of these molecules or manipulate cell behavior, the ability to decorate cell surfaces with bioactive molecules of our choosing is a potentially powerful technique. Here, we describe the molecular engineering of living L6 myoblast monolayers via selective periodate oxidation of sialic acid residues and the application of this surface modification in the artificial aggregation of cells. The aldehyde groups generated by this reaction were used to selectively ligate a model molecule, biotin hydrazide, to the cell surfaces. Flow cytometry analysis after staining with fluorescently conjugated avidin revealed a concentration dependent increase in fluorescence compared to untreated cells with a maximal shift of 345.1 +/- 27.4-fold and an EC(50) of 17.4 +/- 1.1 microM. This mild oxidation reaction did not affect cell number, viability, or morphology. We then compared this chemical technique with the metabolic incorporation of reactive cell surface ketone groups using N-levulinoylmannosamine (ManLev). In this cell line, only a 22.3-fold fluorescence shift was observed compared to untreated cells when myoblasts were incubated with a high concentration of ManLev for 48 hours. Periodate oxidation was then used to modify myoblast surfaces to induce cell aggregation. Crosslinking biotinylated myoblasts, which do not spontaneously aggregate in culture, with avidin resulted in the rapid formation of millimeter sized, multicellular structures. These data indicate that sodium periodate treatment is an effective, noncytotoxic method for the in vitro molecular engineering of living cell surfaces with the potential for cell biology and tissue engineering applications. PMID- 12557314 TI - Solid-phase peptide synthesis by ion-paired alpha-chymotrypsin in nonaqueous media. AB - Solid-phase synthesis of dipeptides in low-water media was achieved using AOT ion paired alpha-chymotrypsin solubilized in organic solvents. Multiple solvents and systematic variation of water activity, a(w), were used to examine the rate of coupling between N-alpha-benzyloxycarbonyl-L-phenylalanine methyl ester (Z-Phe OMe) and leucine as a function of the reaction medium for both solid-phase and solution-phase reactions. In solution, the observed maximum reaction rate in a given solvent generally correlated with measures of hydrophobicity such as the log of the 1-octanol/water partitioning coefficient (log P) and the Hildebrand solubility parameter. The maximum rate for solution-phase synthesis (13 mmol/h g enzyme) was obtained in a 90/10 (v/v) isooctane/tetrahydrofuran solvent mixture at an a(w) of 0.30. For the synthesis of dipeptides from solid-phase leucine residues, the highest synthetic rates (0.14-1.3 mmol/h g-enzyme) were confined to solvent environments that fell inside abruptly defined regions of solvent parameter space (e.g., log P > 2.3 and normalized electron acceptance index <0.13). The maximum rate for solid-phase synthesis was obtained in a 90/10 (v/v) isooctane/tetrahydrofuran solvent mixture at an a(w) of 0.14. In 90/10 and 70/30 (v/v) isooctane/tetrahydrofuran environments with a(w) set to 0.14, seven different N-protected dipeptides were synthesized on commercially available Tentagel support with yields of 74-98% in 24 h. PMID- 12557315 TI - Modeling of yeast metabolism and process dynamics in batch fermentation. AB - Much is known about yeast metabolism and the kinetics of industrial batch fermentation processes. In this study, however, we provide the first tool to evaluate the dynamic interaction that exists between them. A stoichiometric model, using wine fermentation as a case study, was constructed to simulate batch cultures of Saccharomyces cerevisiae. Five differential equations describe the evolution of the main metabolites and biomass in the fermentation tank, while a set of underdetermined linear algebraic equations models the pseudo-steady-state microbial metabolism. Specific links between process variables and the reaction rates of metabolic pathways represent microorganism adaptation to environmental changes in the culture. Adaptation requirements to changes in the environment, optimal growth, and homeostasis were set as the physiological objectives. A linear programming routine was used to define optimal metabolic mass flux distribution at each instant throughout the process. The kinetics of the process arise from the dynamic interaction between the environment and metabolic flux distribution. The model assessed the effect of nitrogen starvation and ethanol toxicity in wine fermentation and it was able to simulate fermentation profiles qualitatively, while experimental fermentation yields were reproduced successfully as well. PMID- 12557316 TI - Integrated optical sensing of dissolved oxygen in microtiter plates: a novel tool for microbial cultivation. AB - Microtiter plates with integrated optical sensing of dissolved oxygen were developed by immobilization of two fluorophores at the bottom of 96-well polystyrene microtiter plates. The oxygen-sensitive fluorophore responded to dissolved oxygen concentration, whereas the oxygen-insensitive one served as an internal reference. The sensor measured dissolved oxygen accurately in optically well-defined media. Oxygen transfer coefficients, k(L)a, were determined by a dynamic method in a commercial microtiter plate reader with an integrated shaker. For this purpose, the dissolved oxygen was initially depleted by the addition of sodium dithionite and, by oxygen transfer from air, it increased again after complete oxidation of dithionite. k(L)a values in one commercial reader were about 10 to 40 h(-1). k(L)a values were inversely proportional to the filling volume and increased with increasing shaking intensity. Dissolved oxygen was monitored during cultivation of Corynebacterium glutamicum in another reader that allowed much higher shaking intensity. Growth rates determined from optical density measurement were identical to those observed in shaking flasks and in a stirred fermentor. Oxygen uptake rates measured in the stirred fermentor and dissolved oxygen concentrations measured during cultivation in the microtiter plate were used to estimate k(L)a values in a 96-well microtiter plate. The resulting values were about 130 h(-1), which is in the lower range of typical stirred fermentors. The resulting maximum oxygen transfer rate was 26 mM h(-1). Simulations showed that the errors caused by the intermittent measurement method were insignificant under the prevailing conditions. PMID- 12557317 TI - Quantitative comparison of transient growth of Saccharomyces cerevisiae, Saccharomyces kluyveri, and Kluyveromyces lactis. AB - A multitude of metabolic regulations occur in yeast, particularly under dynamic process conditions, such as under sudden glucose excess. However, quantification of regulations and classification of yeast strains under these conditions have yet to be elucidated, which requires high-frequency and consistent quantification of the metabolic response. The present study aimed at quantifying the dynamic regulation of the central metabolism of strains Saccharomyces cerevisiae, S. kluyveri, and Kluyveromyces lactis upon sudden glucose excess, accomplished by a shift-up in dilution rate inside of the oxidative region using a small metabolic flux model. It was found that, under transient growth conditions, S. kluyveri behaved like K. lactis, while classification using steady-state conditions would position S. kluyveri close to S. cerevisiae. For transient conditions and based on the observation whether excess glucose is initially used for catabolism (energy) or anabolism (carbon), we propose to classify strains into energy driven, such as S. cerevisiae, and carbon-driven, such as S. kluyveri and K. lactis, strains. Furthermore, it was found that the delayed onset of fermentative catabolism in carbon-driven strains is a consequence of low catabolic flux and the initial shunt of glucose in non-nitrogen-containing biomass constituents. The MFA model suggests that energy limitation forced the cell to ultimately increase catabolic flux, while the capacity of oxidative catabolism is not sufficient to process this flux oxidatively. The combination of transient experiments and its exploitation with reconciled intrinsic rates using a small metabolic model could corroborate earlier findings of metabolic regulations, such as tight glucose control in carbon-driven strains and transient changes in biomass composition, as well as explore new regulations, such as assimilation of ethanol before glucose. The benefit from using small metabolic flux models is the richness of information and the enhanced insight into intrinsic metabolic pathways without a priori knowledge of adaptation kinetics. Used in an online context, this approach serves as an efficient tool for strain characterization and physiological studies. PMID- 12557319 TI - Capture of a recombinant protein from unclarified canola extract using streamline expanded bed anion exchange. AB - The feasibility of applying expanded bed adsorption technology to recombinant protein recovery from extracts of transgenic canola (rapeseed) was assessed. The extraction step results in a suspension of high solids content that is difficult to clarify. The coarse portion of the solids can be removed easily, and our aim was to operate the expanded bed in the presence of the recalcitrant particulates. Recombinant beta-glucuronidase (rGUS) produced in transgenic canola seed was the model system. Diethylaminoethyl (DEAE) and Streamline DEAE resin exhibited similar binding and elution properties for both rGUS and native canola proteins. More than 95% of native canola proteins did not bind to DEAE resins at pH 7.5, whereas the bound proteins were fractionated by two-step salt elution into two groups with the first peak, containing 70% of total bound proteins, at 20 mS/cm, followed by elution of rGUS at 50 mS/cm. The adsorption isotherm was only slightly influenced by the presence of up to 14 mg solids/mL extract; C(m) and K(d) changed by -1% and +39%, respectively. Bed expansion was semiquantitatively predictable from physical properties of the fluid together with Stokes's law and the Richardson-Zaki correlation for both clarified and partially clarified extracts. The presence of 1.4% solids did not change rGUS breakthrough behavior of the expanded bed; however, a small difference between expanded bed and packed bed was observed early in the sample loading stage, during which bed expansion adjusts. Canola solids moved through the column in approximately plug flow with no detriment to bed stability. Seventy-two percent recovery of 34-fold purified rGUS was obtained after initial loading of 1.4% (w/w) solids extract to 25% breakthrough. PMID- 12557318 TI - Human c-fos promoter mediates high-level, inducible expression in various mammalian cell lines. AB - The promoter activity of the human c-fos and human cytomegalovirus (CMV) immediate early promoter was compared in transient and stable transfection experiments with six cell lines of mouse, human, and hamster origin which are all of commercial importance. The c-fos promoter was 1.8-5.6-fold stronger than the CMV promoter in BHK-A, BHK-B, CHO-DHFR(-), and mouse NIH-3T3 in stable transfectants and less effective in mouse myeloma or human 293 cells, suggesting a new transcriptional control element for high-level expression and protein production in mammalian cells. The induction profiles determined in the presence and absence of serum are dependent on the cell line used. Induction levels of up to 8-fold could be achieved in preselected cell pools. PMID- 12557320 TI - On the organic solvent and thermostability of the biocatalytic redox system of Rhodococcus ruber DSM 44541. AB - The sec-alcohol dehydrogenase activity of whole cells of Rhodococcus ruber DSM 44541 has been employed as an efficient biocatalytic redox system due to the use of acetone and 2-propanol at elevated concentrations for cofactor regeneration in the oxidation and reduction mode, respectively, and external addition of NADH/NAD(+) can be omitted. The operational half-life time of the redox system is 29 hours in 20% v/v acetone and 37 hours in 30% v/v 2-propanol. The Redox system allows the enantioselective oxidation of sec-alcohols and the asymmetric reduction of ketones to furnish (S)-configurated alcohols in high optical purity. The stability of the cells towards further organic solvents was investigated. In addition, the system displays thermostability of up to 60 degrees C and pH stability of up to pH 11. The system represents a simple to handle tool for environmentally benign redox reactions. PMID- 12557321 TI - Expression and secretion of the heterodimeric protein interleukin-12 in plant cell suspension culture. AB - It has been suggested that plant cell culture is the most suitable system for producing small-to-medium quantities of specialized, expensive, and high-purity proteins. Here, we report that a heterodimeric protein, human interleukin-12 (hIL 12), was expressed and secreted into culture medium in a biologically active form. A transgenic plant expressing hIL-12 was constructed by sexual crossing of plants that expressed each subunit of the protein. From a piece of transgenic plant, callus was induced and cell suspension culture was established. The biological activity and amount of hIL-12 secreted into culture medium were analyzed using bioassays and ELISA. Analysis of cellular localization demonstrated that the protein was secreted into the culture medium together with its intrinsic signal peptide. PMID- 12557323 TI - [Serum levels of sFas, sICAM-1, IL-18 in patients with chronic hepatitis C and their clinical significance] AB - OBJECTIVE: To investigate the serum levels of soluble Fas antigen (sFas), soluble intercellular adhesion molecules-1 (sICAM-1), interleukin-18 (IL-18) in patients with chronic hepatitis C and to study their roles in pathogenesis of chronic hepatitis C. METHODS: Serum sFas, sICAM-1, IL-18 levels were measured in 30 cases of chronic hepatitis C before and after treatment of interferon-alpha by enzyme linked immunosorbent assay (ELISA), serum titer of HCV-RNA was detected by quantitative PCR and serum ALT activity was also detected. RESULTS: Serum levels of sFas sICAM-1 IL-18 in chronic hepatitis C patients were significantly higher than those in normal controls (P<0.01), showing correlation with serum HCV-RNA titer (r=0.915, r=0.795, r=0.757, respectively, P<0.01), Serum levels of sICAM-1, IL-18 showed correlation with serum ALT level(gamma=0.952, gamma=0.969, respectively, P<0.01), but no relationship was observed between serum sFas and serum ALT level(P>0.05). Serum levels of sFsa sICAM-1 IL-18 markedly decreased in responsive patients while no change was observed in patients with no response after treatment. CONCLUSION: Soluble Fas, soluble ICAM-1, IL-18 may participate in the pathogenesis of chronic hepatitis C and show correlation with the severity of histological inflammation and viral titer. PMID- 12557324 TI - [Expression of mRNA isoforms of vascular endothelial growth factor in ovarian carcinoma] AB - OBJECTIVE: To study the expression of vascular endothelial growth factor (VEGF) mRNA isoforms in ovarian carcinoma and to explore their role in tumorigenesis and development of ovarian carcinoma. METHODS: The types and levels of VEGF mRNA isoforms of surgical samples from 30 patients with ovarian carcinoma were determined by relatively quantative RT-PCR, nest PCR and sequence analysis. RESULTS: VEGF(121), VEGF(145), VEGF(165) and VEGF(189)mRNA were detected in normal ovaries and ovarian carcinoma tissues. The expression level of VEGF(121) was significantly higher than that of VEGF(145), VEGF(165) and VEGF(189) (P<0.001, respectively). The expression of all 4 isoforms in carcinoma tissues was increased significantly compared with that in normal ovaries (P<0.05). CONCLUSION: Overexpression of VEGF(121), VEGF(145), VEGF (165) and VEGF(189) mRNA, especially VEGF(121), was found in varian carcinoma tissues. This findings suggest that all 4 VEGF isoforms may be involved in the tumorigenesis and development of ovarian carcinoma and VEGF(121) may play a key role. PMID- 12557325 TI - [Simvastatin induced apoptosis and its effect on apoptosis-related gene expression in rat vascular smooth muscle cell] AB - OBJECTIVE: To observe the influence of simvastatin on the apoptosis of vascular smooth muscle cells (VSMC) and its effects on the expression of apoptosis-related genes. METHODS: The presence of apoptosis was detected by electron microscope and flow cytometry assessment of PI/Annexin V stain; The protein levels of Bax, Bel-2 and activation of caspase-3 were examined using Western blot technique. RESULTS: After treatment with 30 &mgr;mol/L simvastatin for 24 h, apoptosis were identified with electron microscope in VSMC and flow cytometry showed that rate of apoptosis in simvastatin group (35.5+/-5.8)% was singificantly higher than that in control group (15.1+/-5.0)%. Western blot analyses revealed that the apoptosis process was associated with upregulation of Bax protein and activation of caspase-3, but not with Bel-2 expression. CONCLUSION: Simvastatin can induce apoptosis in VSMC in associated with induction of bax and activation of caspase 3. PMID- 12557326 TI - [Construction of bi-cistronic co-expression plasmid of mIL-12 and the expression in vitro or in vivo] AB - OBJECTIVE: To construct a bi-cistronic co-expression plasmid for mouse interleukin-12 and to observe its expression in vitro or in vivo.METHODS: The full-length cDNA encoding p35 and p40 was cloned into eukaryotic cells expression vector pcDNA 3.1 respectively. Subsequently,the p35 expression unit was inserted into pcDNA 3.1/p40 to produce the bi-cistronic co-expression plasmid in which the p35 and p40 genes were controlled by their own CMV.The plasmid was expressed in vitro and in vivo. RESULTS: The mIL-12 in the supernatant was detected by ELISA after the pCmIL-12 was transfected into COS-7 cells. The activity of NK cells could be augmented by the supernatant in vitro and also by by intradermal delivery of pCmIL-12 in vivo. CONCLUSION: The plasmid constructed by us can express biologically active mIL-12 in vitro and in vivo. PMID- 12557327 TI - [Nitric oxide mediates the inhibitory effects of interleukin-2 on the isolated ventricular papillary muscle and cultured neonatal rat cardiomyocyte] AB - OBJECTIVE: To explore the effects and mechanism of interleukin-2 on the isolated rat ventricular papillary muscle and cultured cardiomyocyte. METHODS: The isolated rat papillary muscle and cultured neonatal rat cardiomyocyte were used to determine the effects on contractile properties and spontaneous beating. RESULTS: IL-2 inhibited the contractile force of isolated rat papillary muscle and reduced the spontaneous beating rate of cultured neonatal rat cardiomyocytes in a dose-dependent manner. IL-2 at high concentration reduced the systolic duration and prolonged the diastolic duration. Pretreatment with L-NAME, an inhibitor of nitric oxide synthase,abolished the effects of IL-2. Heat inactivated IL-2 had no effect on papillary muscles and cardiomyocytes.CONCLUSION: Inhibitory effects of IL-2 on the contraction of isolated papillary muscle and spontaneous beating rate of cultured neonatal cardiomyocyte are mediated by nitric oxide. PMID- 12557328 TI - [Influence of peritoneal dialysate on nitric oxide production and viability of macrophage] AB - OBJECTIVE: To investigate the influence of commercial peritoneal dialysate (CDS) on function of macrophage. METHODS: Cultured peritoneal macrophages were divided into two experimental groups and their controls.(1) Macrophages were cultured in conditioned culture medium containing 50%CDS (0.139 mol/L glucose) for 24 h. (2)Macrophage were exposed to CDS containing 0.139mol/L glucose for 10, 30 and 60 min respectively, and then cultured in CDS-free medium for 24 h. The nitric oxide (NO) production and MTT in two groups were measured. In each control group, CDS was replaced by same amount of culture medium. RESULTS: NO production and MTT reduction ability (related to cell viability) of experimental groups were remarkably lower than those of controls and the NO production and MTT reduction in 60min CDS-exposed group were lower than that of 10 min and 30 min CDS-exposed group. CONCLUSION: Dialysate may have detrimental effects on viability and other function of macrophage and these effects may related to time length of CDS exposure. PMID- 12557329 TI - [Experimental study on allogeneic mandible transplantation combined with basic fibroblast growth factor] AB - OBJECTIVE: To study the effect of basic fibroblast growth factor(b-FGF) on revascularization and bone remodeling of allogeneic mandible transplantation in repair of mandible defects in rabbits. METHODS: The mandible defects of 20 adult rabbits were created in both sides. The defects on the left side were implanted with allogeneic bone and local administration of b-FGF; the defects on the right side were only repaired with allogeneic bone as control group. At 1, 3 months after operation, the revascularization and bone remodeling were observed by ink gelation vascular perfusion-transparency and histological examination. RESULTS: The allogeneic bone and b-FGF group had more marked vascularization and more quick and complete bone formation than control group. CONCLUSION: b-FGF can improve revascularization and bone formation after allogeneic mandible transplantation; allogeneic bone combined with b-FGF is a promising bone substitute in clinical uses. PMID- 12557330 TI - [High bioactive material-bone graft in bone defect healing] AB - OBJECTIVE: To assese the value and clinical significance of bioactive material hydroxyapatite (HA) combined with TGF-beta1 in bone defect healing. METHODS: The applicability of bioactive material-HA granules combined with TGF-beta1 (group A) as a substitute for bone graft was observed in SD rat model. The results were compared with those of HA granules (Group B) and ungrafted bone defect (Group C). RESULTS: Group A demonstrated the highest level of type I collagen mRNA during healing period. CONCLUSION: The bioactive material-HA combined with TGF-beta1 can efficiently bond to ongrowing new bone comparing to HA granules. PMID- 12557331 TI - [Effects of liposomal prostaglandin E(1) on myocardial reperfusion injury in rabbits] AB - OBJECTIVE: To probe into the role of liposomal prostaglandin E(1) (Lipo-PGE(1)) on reperfusion injury in a rabbit ischemia-reperfusion model. METHODS: Twenty four open-chest rebbits were randomized to receive a 10 min intravenous infusion of either liposome diluent, free PGE(1), or Lipo-PGE(1) after 60 min of left anterior desending (LAD) ligation just before reperfusion. Serum creatine phosphokinase (CPK), malodialdehyde (MDA), superoxide dismutase (SOD) were detected; infarct size and region at risk were measured. RESULTS: Infarct size as a ratio of weight of infarcted tissue to weight at risk (MI/RISK) was significantly reduced with Lipo-PGE(1) (32.20+/-4.70)% compared with PGE(1) (42.09+/-6.93)% or placebo (44.57+/-5.46)% infusion (P<0.01). The values of serum CPK, MDA during reperfusion in treatment of Lipo-PGE(1) group were significantly reduced than in treatment of PGE(1) group or control group (P<0.05), and the values of serum SOD were significantly increased (P<0.05). CONCLUSION: Lipo PGE(1) can effectively decrease the serum CPK and MDA contents, elevate the SOD activity, and attenuate myocardial reperfusion injury. PMID- 12557332 TI - [Effect of adriamycin on sarcoplasmic reticulum Ca2+ -ATPase activity in cardiomyocyte of rabbits] AB - OBJECTIVE: To evaluate influences of regular-dose of adriamycin (ADR) on heart function and sarcoplasmic reticulum (SR) Ca2+ -ATPase in cardiomyocyte of rabbits. METHODS: Nine rabbits received intraveneous injection of ADR (2mg/kg) once a week for 8 weeks, the rabbits injected with saline were used as control group. Cardiac output (CO), blood pressure (BP), mean artery pressure (MAP), left ventricular systolic pressure( LVSP), left ventricular end-diastolic pressure (LVEDP), calcium in cardiomyocyte (MyoCa2+) of rabbits and SR Ca2+ -ATPase activity were examinated 3 weeks after the final injection. RESULTS: CO, LVSP and SR Ca2+ -ATPase activity were significantly decreased in ADR treated group compared with the control group. Conversely, LVEDP and MyoCa2+ were significantly increased in ADR treated rabbits. CONCLUSION: Heart function can be decreased by regular-dose of ADR in injection. Calcium overload in cardiomyocyte and decrease of SR Ca2+ -ATPase activity is important physiopathologic mechanism in ADR induced impairment of heart. PMID- 12557333 TI - [Relation between formation time and structure of red blood cell aggregation body] AB - OBJECTIVE: To study the relation between of forming time red blood cell aggregation body and structure by changing the thickness of flow channel plane and red blood cell hematocrit. METHODS: A system with flow channel technique and light transmission measurements were applied to gaining red blood cell aggregation curve. RESULTS: The forming time of red blood cell aggregation body and the thickness of aggregation body as well as red blood cell hematocrit were linearily correlated with a correlation coefficient gamma of 0.989 and 0.990 respectively. CONCLUSION: When red blood cell aggregation process is studied using different methods, red blood cell aggregation time index can not be compared. PMID- 12557334 TI - [Preparation of albendazole sulfoxide--- an active metabolite] AB - OBJECTIVE: To investigate the facile method for preparation of albendazole sulfoxide, an active metabolite in vivo. METHODS: The reaction condition was investigated according to evaluations to the results of the experiment through temperature, solvent, oxidation agent, reaction time and impurity. RESULTS: The target compound was obtained by oxidation of albendazole with the presence of sodium metaperiodate in glacial acetic acid under low temperature and was free from contamination by impurity albendazole sulfone. Pure product (content >98%) and high yield (>90%percnt;) have been achieved due to reaction selection. CONCLUSION: Oxidation condition is essential to the preparation of albendazole sulfoxide. PMID- 12557335 TI - [Study on application of PCR in the diagnosis of Haemophilus influenzae pneumonia] AB - OBJECTIVE: To study the role of Haemophilus influenzae(Hi) in pneumonia and that of PCR in the diagnosis of Hi pneumonia. METHODS: Hi genus-specific PCR, Hib type specific PCR and selective Hi culture media were used to detect 83 samples of deep nasal pharyngeal aspiration (NPA), 51 sera from 83 children younger than 3 years with pneumonia and 37 samples of pharyngeal swabs from healthy children. RESULTS: Of 83 NPA samples, 20(24.1 %) were positive by culture, 36 (43.4 %) positive by Hi-PCR and 19 (22.9 %) positive by Hib-PCR.Six out of 51 sera were positive by Hi-PCR and Hib-PCR, but none positive by culture. Of 37 pharyngeal swabs from healthy children, 3 ( 8.1 %) were positive by culture, 6 ( 16.2 %) positive by Hi-PCR and none positive by Hib-PCR. CONCLUSION: Hib-PCR is more appropriate for detecting NPA samples from children with pneumonia because of the high rate of non-typeable Hi carriers in healthy children. PMID- 12557336 TI - [Serum interleukin-6 levels of appendicitis in children] PMID- 12557337 TI - [Treatment of hypospadias by mobilization of the anterior urethra and meatal advancement] PMID- 12557338 TI - [Transcatheter closure of patent ductus arteriosus with Rashkind occlude and Amplatzer duct occluder] PMID- 12557339 TI - [Diagnosis and surgical treatment of Crohn's disease] PMID- 12557340 TI - [Modulation of effects of norepinephrine on the hear] PMID- 12557341 TI - [Study on the reclassification of two deposited strains AS4.1186 and AS4.1187 of Nocardia]. AB - Polyphasic evidence for the reclassification of Nocardia nostocoides AS4.1186 and Nocardia salmonicolor var. aurantiaca AS4.1187 indicated that strain AS4.1186 was closely related to Saccharothrix texasensis and strain AS4.1187 was very closely related to Rhodococcus ruber. Binary 16S rDNA sequence for the pair AS4.1186 and Saccharothrix texasensis NRRL B-16134T, and the pair AS4.1187 and Rhodococcus ruber DSM 43338T exhibited 99.3% and 99.5% similarity, respectively. Corresponding DNA-DNA reassociation values were significantly higher than 70%, 77.6% and 82.9%, respectively. Results of chemotaxonomic analyses of cell wall, mycolic acid, principal menaquinones, phospholipid type and the G + C content of the DNA supported the conclusions of the genotypic analyses. The very similar morphological and physiological characteristics agreed with the high degree of relatedness. On the basis of phylogenetic analyses, DNA-DNA reassociation values, chemotaxonomic properties, morphological and physiological characteristics, it is concluded that strain AS4.1186 and AS4.1187 should be removed from the genus Nocardia. We propose to transfer Nocardia nostocoides AS4.1186 to the genus Saccharothrix, combine with the species S. texasensis, and combine Nocardia salmonicolor var. aurantiaca AS4.1187 with the species Rhodococcus ruber. PMID- 12557342 TI - [Tn4560 transposition in Streptomyces ansochromogenes and its application for cloning of genes involved in nikkomycin biosynthesis]. AB - No transformant was obtained when pUC1169(pMT660::Tn4556::vph) from S. coelicolor J1501 was used to transform the protoplast of S. ansochromogenes by general transformation for many times. Transformants were obtained when the protoplast of S. ansochromogenes was prepared at 50 degrees C for 30 min by heat attenuation of restriction. But the transformation frequency is very low, only 0.4 transformant per microgram DNA. Transformation frequency increased 10(3)-10(4) times when the protoplast of S. ansochromogenes was transformed by pUC1169 from itself. With Tn4560 transposition, 8 mutants lost ability to produce nikkomycin were screened from 4068 colonies with VioR-ThioS. Southern blotting suggested that Tn4560 was inserted in at least 4 positions of chromosomal DNA in S. ansochromogenes. By using the cloned 30 kb DNA fragment involved in nikkomycin biosynthesis as a probe, southern hybridization was carried out with 8 mutant total DNA digested with different enzymes. Only total DNA from Nik5 displayed signals, which were the same as those of the wild type. With HPLC analysis, 8 mutants could be classified into two types for nikkomycin production, one is Nik5, the other is Nik1, 2, 3, 4, 6, 7 and Nik8. As deduced preliminarily, the deletion of large chromosomal DNA fragment involved in nikkomycin biosynthesis took place in the 7 mutants. The partial gene library of Nik5 was constructed, the 4.2 kb and the 0.6 kb chromosomal DNA fragment flanking Tn4560 were cloned. Sequence analysis showed that the 4.8 kb DNA fragment (GenBank accession number:AF349618) contains two complete ORFs and one partial ORF. PMID- 12557343 TI - [Isolation and characterization of a p-nitrophenol degradation Pseudomonas sp. strain P3 and construction of a genetically engineered bacterium]. AB - A Pseudomonas strain P3 was isolated in this work. P3 can grow with p-nitrophenol as the sole carbon and nitrogen source. Growth in media with nitrogen, P3 can degrade p-nitrophenol to accumulate nitrite in the culture media. P3 can utilize a series of aromatic compounds as sole carbon sources. Different heavy metal ions have different effects on the degradation of p-Nitrophenol by P3. Glucose had no effect on the degradation of p-nitrophenol, while trace yeast extract greatly increased the degradation rate. Methyl parathion hydrolase gene mpd was clone into P3 by conjugation and genetically engineered bacterium PM was obtained. Methyl parathion hydrolase was expressed by PM. PM could grew on methyl parathion as sole carbon source. PM could degrade methyl parathion with relatively high activity and stability. PMID- 12557344 TI - [Construction of hpaA gene-engineered attenuated Salmonella typhimurium vaccine strain]. AB - To express Helicobacter pylori hpaA gene in attenuated Salmonella typhimurium vaccine vehicle, and elucidate the potential value of attenuated Salmonella typhimurium as a vector expressing Helicobacter pylori antigens, by means of molecular biology, 783 bp hpaA gene was cloned into NcoI-SalI site of a procaryotic expression plasmid pTrc99A, and the recombinant plasmid was then used to transform an attenuated Salmonella typhimurium vaccine strain SL3261, and the positive clones were screened by PCR and restriction enzyme digestion. HpaA expression was analyzed by SDS-PAGE and Western blot. Two and 10 days after recombinant strain intragastric immunization, the C57BL/6 mice was sacrificed, and the spleen and terminal ileum was cultured for recombinant strain. The results showed that a recombinant procaryotic expression plasmid pTrc99A-hpaA was constructed, and the recombinant plasmid was then introduced into an attenuated Salmonella typhimurium vaccine strain SL3261 successfully. HpaA was expressed in the recombinant strains as a 30 kD protein, and also its immunogenicity was confirmed by Western blot. Recombinant strain was found in both spleen and terminal ileum of each mouse two and ten days after intragastric immunization. We concluded that a recombinant live attenuated Salmonella typhimurium vaccine strain expressing Helicobacter pylori hpaA gene was constructed and identified, and this work will help to develop oral recombinant live vaccine strains against Helicobacter pylori infection. PMID- 12557345 TI - [The cloning and functional analysis of Sinorhizobium fredii 042BS regulatory modulation genes]. AB - The total DNA of Sinorhizobium fredii 042BS was digested by EcoRI for Southern blotting with probes of nodD1 and nodD2 from S. fredii USDA257. The 3 kb positive band hybrided with nodD1 probe and 6 kb positive band with nodD2 probe were found, respectively. Partial gene library were constructed using pUC18 as vector, and the clones with the nodD1 and nodD2 genes were obtained. The sequence of nodD1 and nodD2 of 042B showed that they are highly homologous with nodD1 and nodD2 of S. fredii. The fragment with nodD1 was cloned into the vector pBBRIMCS-5 and introduced into R. leguminosarum bv. viciae LPR5054 to study the function of the nodD1. The results showed that nodD1 of 042B can be induced by genistein and luteolin secreted by the seedlings of soybean and alfalfa respectively. PMID- 12557346 TI - [The study on cloning and expression of Bt cry1Ab13 gene]. AB - B. thuringiensis strain C005 with high insecticidal activity to several kinds of pests, screened from China, was identified that it contained cry1Ab gene by PCR RFLP. Southern blotting showed that a 8.5 kb positive band of plasmid DNA digested with PstI contained cry1Ab gene. The gene was cloned from Bt C005 and the results of sequence analysis showed that cry1Ab13 gene contained a 3468 bp open reading frame, encoding a 130.6 kD protein composing 1155 amino acids. The IE point of Cry1Ab13 protein was pH 4.845. The cry1Ab gene has been registered in GenBank (Accession number is AF254640) and named as cry1Ab13 as a novel gene by International Nomenclature Committee of Bt delta-endotoxin genes. SDS-PAGE analysis indicated that 130 kD protein of Cry1Ab13 was expressed in a Bt acrystalliferous mutant cryB- and bioassay results proved that the transformant BiotI81 containing cry1Ab13 gene had high toxicity to Plutella xylostella. PMID- 12557347 TI - [The cloning of plasmid replicon ori165 from Bacillus thuringiensis subsp. thenebrionis]. AB - An EcoRI DNA fragment containing the plasmid replicon, named ori165, was cloned from the plasmids of B. thuringiensis subsp. tenebrionis strains YBT-1765. The replicon ori165 was the first replicon cloned from this subspecie. Its size is approximately 11 kb. The replicon was minimized to an 8 kb fragment by deletion analysis. Southern hybridization exhibited that ori165 was coming from the plasmid detected with the highest molecular weight. A shuttle vector pBMB6071 based on ori165 was constructed. The stabilities of pBMB6071 in different B. thuringiensis host had great discrepancy. In strain 4Q7, a crystal-negative derivative of B. thuringiensis subsp. isrealensis, the stability was 100%. ori165 was compatible with ori1030 and ori2062 in strain BMB171, a crystal-negative derivative of B. thuringiensis subsp. kurstaki. PMID- 12557348 TI - [Package of Bombyx mori nuclear polyhedrosis virus with heterogoneous polyhedring protein]. AB - Cloning the segment of polyhedrin gene which was separated from AcMNPV genome DNA by PCR method into transfer vector pBacPAK8, we got recombinant transfer vector pOAc, then contransfecting BmN cells with the vector and linear virus Bm-BacPAK6, harvested recombinant virus hp-BmNPV which can produce polyhedrin but no blue plaques. Analysing of recombinant virus from polyhedrin, recombinant viral DNA and polyhedra, we confirmed that polyhedring of AcMNPV can be not only high-level expressed in BmN cells, But can recognize recombinant BmNPV DNA and assemble them to polyhedra. We observed that the pattern of restriction enzyme changed due to DNA recombination, the size of recombinant viral polyhedron (1.2-2.9 microns) was obviously smaller than those of wild-type BmNPV by electron microscopy. PMID- 12557349 TI - [Amplification of bacterial heat shock protein 60 gene using inverse PCR method]. AB - A method is presented for rapid in vitro amplification of DNA sequences of bacterial HSP60 gene. In our previous work, using degenerate oligonucleotide primers for conserved regions of HSP60 gene, a 600 bp fragment was amplified by PCR, cloned and sequenced. An inverse PCR method, with the primers oriented in the reversed direction of the usual orientation, is used to amplify the DNA sequences that flank the 600 bp known region. The feasibility of this method is shown by amplifying the complete HSP60 gene sequences of 4 bacterial strains, namely Bifidobacterium denticolens, Bifidobacterium inopinatum, Bifidobacterium adolescentis and Gardnerella vaginalis. PMID- 12557350 TI - [Expression of recombinant human serum albumin in genetically engineered Pichia pastoris in high-density fermentation]. AB - The optimum culture conditions of genetically engineered Pichia pastoris in shake flask cultivation and in fed-batch fermentation were investigated respectively in this paper. It showed that the cultural period induced with 5 g/L methanol is 96 h, optimum methanol concentration is 10 g/L, and pH range is 5.72-6.59 in shake flask culture. Although the seed inoculum amount increased, the target protein productivity per cell optical density was decreased. Their relationship fit the equeation Y = 12.941x(-0.5059) (r = 0.9789, where x is inoculating OD600, Y is protein productivity per cell optical density), we postulated that the restricting factor may be dissolved oxygen (DO) at shake-flask culture. With the 10% inoculum and 20 OD600 of seed, the lag phase of cell growth is 2.11 h in batch cultivation, and the relationship between cell optical density (Y) and culture time (t) is Y = 0.7841e0.2319t (r = 0.9936); The cell dry weight of broth reached 115-160 g/L and the maximum rHSA concentration was 3.6 g/L at the 120th at the fed-batch fermentation phase. PMID- 12557351 TI - [Study on detection of the Mycobacteria DNA in formalin-fixed, paraffin-embedded tissue samples by triplex polymerase chain reaction]. AB - To supply an additional differential diagnostic method for pathological diagnosis of Mycobacterium tuberculosis complex and nontuberculous mycobacteria infections in formalin-fixed, paraffin-embedded tissue samples by triplex-PCR. Three pairs of oligonucleotide primer were used in triplex-PCR. A 383 bp DNA fragment encoding part of the 65 kD mycobacterial surface antigen, a 123 bp fragment corresponding to a specific Mycobacterium tuberculosis complex sequence which was the insertion sequence 6110 (IS6110) and a 268 bp fragment for human beta-globin were amplified by triplex-PCR respectively. The sensitivity of the triplex-PCR electrophoresis for the mycobacteria DNA was 0.6 picogram. The specific bands of 383 bp and 123 bp among the amplified DNA from Mycobacterium tuberculosis, M. bovis, M. bovis BCG and M. simiae were present in the agarose gel. By contrast, only a band of 383 bp was found among the nontuberculosis mycobacteria which contained M. avium, M. chelonae, M. scrofulaceum, M. xenopi, M. kansasii, M. intracellulare and M. smegmatis. Compared with the standard strains, there was an additional 268 bp band in simulated clinic samples infected by mycobacteria, 209 formalin-fixed, paraffin-embedded tissue samples of the patients diagnosed as scrofula by clinic doctor at first visit were examined by triplex polymerase chain reaction. Among them, 193 tissue samples of the patients pathologically diagnosed as scrofula, tuberculous granulomatous tissue or tuberculous granulomatous inflammation were positive: the specific hands of 383 bp, 123 bp and 268 bp were present in the agarose gel and this tallied with Mycobacterium tuberculosis complex infection. Of 16 tissue samples of the patients pathologically diagnosed as suspicious scrofula, 15 samples were same positive results and this tallied with Mycobacterium tuberculosis complex infection, too; 1 sample could find the specific bands of 383 bp and 268 bp which were present in the agarose gel and this tallied with nontuberculous mycobacteria infection. The results showed that the triplex-PCR could detect and identify the DNA of Mycobacterium tuberculosis complex and nontuberculous mycobacteria except M. simiae. It is a valuable detecting method which has high sensitivity and specificity. PMID- 12557352 TI - [Studies on enzymatic synthesis of cephalexin by immobilized penicillin acylase by polyacrylonitrile fibres]. AB - The extracellular penicillin acylase from Bacillus megaterium was immobilized by coupling to derivatives of polyacrylonitrile fibres. The apparent activity of the immobilized enzyme was about 153 U/g (wet weight). The optimal pH and temperature were 6.5 and 40 degrees C for synthesis of cephalexin by penicillin acylase, respectively. Whe the concentration of 7-ADCA was 4% and the ratio of PGME and 7 ADCA and 1:2, the average velocity of synthesis reaction was highest. The optimal supplied amount of immobilized penicillin acylase was 1.125 g/g 7-ADCA.. The apparent Michaelis constant for 7-ADCA was 0.162 mol/L and for PGME was 0.364 mol/L, Vmax was 0.0462 mol.L-1.min-1 at 30 degrees C and pH6.5. The remained activity was about 83.9% after operating 50 times. PMID- 12557353 TI - [Production of D-pantolactone hydrolase by Fusarium moniliforme SW-902]. AB - Fusarium moniliforme SW-902, a high-yielding strain for producing D-pantolactone hydrolase was screened. The conditions for enzyme formation of the strain were studied. The suitable carbon source of the medium was glycerol and the suitable nitrogen source was peptone. The optimum temperature and initial pH for enzyme formation were 26 degrees C and pH8.0 respectively. When the organism was cultured in the 60 L-fermentor under the optimum conditions for 45 h, about 7.18 g dry cells/L and 0.92 IU/g dry cells weight were obtained. PMID- 12557354 TI - [Purification and some properties of D-carbamoylase]. AB - A D-Carbamoylase produced by a strain NO. 2262 was purified to electrophoretic homogeneity with the recovery of 20% activity and the purification factor of 8 fold by three steps including (NH4)2SO4 fractionation, hydrophobic column and pre packed Hitrap Q HR. It is indicated from the results of nativ-PAGE and SDS-PAGE analysis that the enzyme could be a homogeneous tetramer consisting of four 35 kD subunits. In addition, its optimal pH and optimal temperature are 8.0 and 45 degrees C respectively. The basic kinetic parameters of the enzyme are Km = 1.3 x 10(-3) mol/L and Vmax = 0.33 mumol/min with N-carbamyl-DL-Alanine as the substrate. The effect of bivalent metal ions on the enzyme was showed that Ni2+ could be as an activator, Zn2+ as a powerful inhibitor, while Co2+ had no any influence at all. Its N-terminal sequence is TRQKILAF in turn. PMID- 12557355 TI - [The kinetic characteristics of mycelium growth of medicinal mushroom Ganoderma sinense in liquid-submerged cultivation]. AB - The study was carried out to investigate the kinetic characteristics of Ganoderma sinense mycelium growth in liquid-submerged cultivation, using a air-lift bioreactor ALR/ff specially designed for the cultivation of mushrooms' suspending mycelia cultures. Mycelia of G. sinense grew in the range of 25 degrees C-35 degrees C, and at the different temperature in which the specific growth rate showed by the mycelia much differed. The specific growth rates with the maximum of 0.0444(h-1) at 1.64 vvm of aeration in the cultivation were more than at 0.93 vvm, also more than at 2.50 vvm. In comparison with the less glucose concentration, the culture of 2.80 g glucose/100 mL in the medium had a shorter lag-period, and the more concentration of glucose favoured mycelia growth in the latter cultivation course with the denser pellets of mycelia. Then the culture of the less concentration of glucose in the medium had the higher conversion-rates of glucose to G. sinense biomass all along the cultivation course, than of the more concentration of glucose. Futhermore, in the continuous cultivations with growth-limiting substrate of carbon source the mycelia displaying different growing activities, demonstrated the varied forms of mycelium-gathering. In that cases the relationships among mycelia biomass, growth-limiting substrate and productivity of mycelium biomass obviously differed from the ones in the case of bacteria. Also in the range of dilution rates 0.010-0.220 (h-1) the growing of G. sinense mycelia followed the function of Contois equation. PMID- 12557356 TI - [Antigen analysis of envelope gene products of avian leukosis virus subgroup J]. AB - Envelope glycoprotein of avian leukosis virus Subgroup J (ALV-J) determines the host range of virus infection and cross-neutralization patterns. The truncated envelope genes of avian lecukosis virus subgroup J (ALV-J) were amplified by PCR and cloned them into pGEX-5X-3 vector for expressing envGST-fusion protein. Western blot analysis results showed that the products of truncated env gene expressed in Escherichia coli could reacted with G2, JE9 and I45 monoclonal antibodies (Mabs) specific to envelope protein of ALV-J. Using different Mabs to map the epitopes in the expressed truncated gp85 GST fusion protein, the results showed that Mab G2 and JE9 antibodies recognizing epitope in gp85 was localized between amino acid 65-155. Mab I45 reacted with the epitope at the location of amino acid 156-233. It indicated that the specificity of subgroup J virus is determined by the gp85 peptide since GST-gp85 protein expressed in Escherichia coli is not glycosylated. PMID- 12557357 TI - [Analysis of virulence-related proteins of Streptococcus suis type 2 from swine Streptococcus isolated in China]. AB - Virulence-related proteins, muraminidase-released protein (MRP) and extracellular factor (EF) of Streptococcus suis type 2, were extracted from Jiangsu Isolate HA9801 and were used as antigens for preparation of antibodies. Bacterium cell envelope proteins and extracellular proteins of swine Streptococcus strains including 17 Chinese isolates, 1 German strain and 1 human isolate of Streptococcus suis type 2, were analyzed by SDS-PAGE and immunoblotting using the above antibodies, 11 strains produced MRP and 10 strains possessed EF or EF. They exist four phenotypes: MRP+ EF+ (8/19), MRP+ EF (1/19), MRP+ EF- (1/19), MRP- EF- (10/19). PMID- 12557358 TI - [Cloning of flocculent gene and expression in industrial yeast strain]. AB - The partial genomic library of Saccharomyce cerevisiae FL189 possessing strong flocculation ability was constructed using Yeast-E. coli shuttle plasmid YCp50 as vector. Recombinant plasmid containing flocculation gene was obtained by screening the growth of transformants on the selective medium and measurement flocculation, designated as pCF1.pCF1 was introduced into industrial yeast strain PJ208-5-15. Six transformants PJ208-5-15-1(pCF1)-PJ208-5-15-6(pCF1) possessing strong flocculation ability were obtained. The results of Southern blot and restriction endonuclease analysis showed that the insert is about 4.3 kb and could hybridize with the probe (2.6 kb Eco RV fragment of FLO1). Flocculation ability assay indicated that the transformants possess strong flocculation ability. Hence, the gene controlling flocculation phenotype exists in the cloned DNA fragment. The restriction endonuclease analysis and the sequence analysis of the insert DNA fragment are in progress. PMID- 12557359 TI - [Study on fermentation of n-paraffin for producing mixed dicarboxylic acids]. AB - A mutant of Candida tropicalis FYD-2 was obtained from its parental strain SFP 1186 by ultraviolet treatments. On shaking flask, the yield of mixed dicarboxylic acid(DCA) by the mutant was 21.4% higher than that by its ancestor. The amount of mixed DCA reached 156 g/L for 120 h incubation in a 10 L autoconrolled fermentor where the culture medium contained 25% n-paraffin. The process of induced and screening mutant was introduced and the time course of fermentation in 10 L fermentor was discussed. PMID- 12557360 TI - [Studies on biotransformation of 5-FUR]. AB - Conditions for biotransformation and purification of FUR were investigated. The result showed that when the cell concentration of E. aerogenes was 10% (w/v), the temperature and pH were 7.8 and 60 degrees C respectively, 59.7% UR was converted to FUR. It also demonstrated that the ideal procedure for the purification of FUR is silica gel column chromatography by two elution systems (S1:CHCl3:CH3OH:H2O = 61:13:1, and S2; CH3COOCH2CH3:CH3OH:CH3COOH:H2O = 12:3:3:2) with the purity and yield of 98.0% and 86.0% respectively. PMID- 12557361 TI - [Genetic modification of bacteria for bioremediation]. PMID- 12557362 TI - [Metabolism and beta-oxidation of alkane-utilizing Candida tropicalis]. PMID- 12557363 TI - [Effects of H2O2 addition on oxygen supply and metabolism of microorganisms]. PMID- 12557364 TI - [Classification for one pathogenic Vibrio anguillarum strain isolated from skin ulcer flounder]. AB - A pathogenic bacterial strain M3 revealed gram negative rod shape, motile and translucent clone was studied. This bacterium was isolated from the skin-ulcer flounder in the fish farm of RongCheng City, Shandong Provice, and could not be identified by BIOLOG ID SYSTEM. Comparative 16S rDNA sequence analyses revealed that strain M3 was most related to the genus of Vibrio. The overall similarity value between strain M3 and Vibrio species were 94% to 98%. Phylogentic analysis showed that M3 strain exhibited the highest level of similarity to Vibrio anguillarum, and the biological features of strain M3 were very similar to that of Vibrio anguillarum. Based on the results obtained, strain M3 was identified as Vibrio anguillarum. PMID- 12557365 TI - [Isolation and screening of beneficial endophytic bacteria to control bacterial ring rot of potato]. AB - One hundred and thirty-three bacterial strains were isolated from inner tissue of potato tubers collected from Datong, Taiyuan and Inner Mongolia Autonomous regions. On the basis of antagonistic examination in vitro, greenhouse and field tests, five strains named as 069, 085, 110, 116 and 118 were chosen for their suppression of bacterial ring rot or their growth promotion. Strain 118 is an endophytic bacterium with three effects of colonization, growth promotion and suppression of the pathogenic bacteria, showing good prospects for commercial use. PMID- 12557366 TI - [Proteomic analysis of different fractions in M. tuberculosis culture]. AB - To provide a basis of molecular genetic analysis of M. tubereulosis, the proteomic profiling was prompted. M. tuberculosis H37RV was cultured in Sauton medium at 37 degrees C for 3 weeks, harvested and fractionated into three portions: suspension filtration proteins(A), cytosol proteins(B) and membrane proteins(C). These fractions were analyzed by pH3-10 IPG gradient and SDS-PAGE. The silver-stained technigue and gel images were used. Then the image was transfered into 2-DE gel analysis Software. A part of protein sports of expression level higher from the culture filtrate fraction were identified by peptide mass fingerprinting. A total of 907, 884 and 681 protein sports were observed for A. B. C fractions in M. tuberculosis H37RV, respectively. Distribution of proteins mass for 3 fractions were principally similar, About 70.5-74.4 per cent were distributed in the ranges of Mr 10-49 kD.pI of the proteins for A, B fractions were pricipally similar, About 80.9-83.5 per cent were distributed in the ranges of pH 3.0-6.4, But the number of protein sports for C fraction distributed in the ranges of pH 7.6-10.0 were more than A, B fractions. The number of protein sports of expression level higher for A, B and C fractions were 71(7.8%), 242(27.4%), 19 (2.8%), respectively. 90 pen cent from them, pH of the proteins were distributed in the ranges of pH 3.0-6.4. 73.1 per cent for proteins mass of C fraction were distributed in the ranges of 10 kD-49 kD, which were more than A, B fractions. Nine of the proteins identified ih this study appeared to be homology or putative fanction proteins, but another five proteins were unknown. The proteomic profiling of different fractions in M. tuberculosis obtained in here will be provide a basis for detailed analysis of biology functions of the proteins. PMID- 12557367 TI - [Cloning, expression of the gI gene of pseudorabies virus ea strain and its effection to viral replication]. AB - The complete gI was cloned from the genomic DNA of Pseudorabies virus (PRV) Ea strain and the DNA sequence was determined by Sanger's sequencing technique. The nucleotide and deduced animo acids sequences indicated that the gI gene of PRV Ea strain is composed of 1101 base pairs and could encode 366 animo acids residues. The result of predicted II class structure showed that gI is a typical I class envelope protein. When compared with PRV Rice strain, there were multiple mutations in the gI gene of PRV Ea strain, and the diversity of amino acid residues also existed, especially the deletion of two bases which results in the change of the reading frame and there were additional 16 animo acids residues in the potential cytoplasmic domains of Ea strain. The fragment containing the complete gI gene was further sub-cloned into the downstream of CMV promoter of eukaryotic expression vector pcDNA3.1+, resulting in a recombinant expression plasmid pSB209. pSB209 was transfected into PK-15 cell lines and the cells expressing gI were obtained in the selection of G418. Expressed protein was further detected by Indirect Immunofluorescence Assay(IFA). In order to explore the function of gI in the viral replication, the attenuated live vaccine strain, Bartha in which the gI is deleted, was chosen to determine the plaques forming units (pfu) and the tissue culture infectious dose(TCID50) in PK-15 cell lines expressing the gI(PKgI). The data showed that the pfu and TCID50 in PKgI were 164% and 200% of those in control cell lines respectively. The above results indicated that the gI involve in and accelerate the viral replication. PMID- 12557368 TI - [Comparison of sequences of the hemagglutinin gene and phylogenetical analysis of H9 subtype avian influenza viruses isolated from some regions in China]. AB - In order to explore the genetic mutaions of the hemagglutinin(HA) gene and the law of molecular epidemiology of H9 subtype avian influenza viruses in China, 23 H9 subtype avian influenza viruses(AIVs) were isolated from 12 provinces of China in recent years. Their nucleotide sequences of cDNA of HA gene were determined by RT-PCR and sequencing. Their nucleotide and putaive amino acid sequences homology was compared. The results showed that their nucleotide sequence homology was from 94.1% to 100% and that amino acid sequence homology was 95.4% to 100%. The sequences of the HA gene of these isolates were analyzed and compared with that of another 8 isolates from reference. The similedty indicated that HK170499 isolated from Hong Kong was close to the 2 isolates of Japan. And of the 31 isolates with complete HA gene sequences there were 5 isolates, HA gene of which were loss of one potelltial glycosylation site, which were CKGS199, CKTJ196, CKT296, CKSH300 and CKBJ197. Then 1098 nucleotide regions (bases 55 to 1,152) of HA gene of 23 isolates in this study were analyzed phylogenetically and compared with sequences from 31 H9 subtype viruses available in the GenBank database. Although considerable variation at the cleavage sites of the different viruses was observed, giving 10 different amino acid motits, none had multiple basic amino acids that correlate with highly pathogenic avian influenza (HPAI) isolates. Examination of amino acid sequences involved in repeptor binding site(RBS) revealed that the amino acid residue at position 191 characteristically distributed in the 54 isolates, that is, this amino acid residue of the isolates of mainland China and several Hong Kong strains was Asn(N) and that of the others was His(H). And the 141 143 amino acid residues, involved in forming the potential glycosylation sites, had the similary characteristic distribution with the 191aa position. The isolates with Asn-191. excluding CKBJ197, had NVS in the position 141aa143aa, meanwhile those with His-191 had NVT. Twenty-six mainland China isolates was genetically in Eurasian lineage but did not show distinctly geographical and temporal relationship. It is concluded that in recent years H9N2 subtype AIV, circulating in chicken flocks of mainland China, may have a common origini. These findings provides importan basis for establishment of scientifically preventive measurements to control H9 subtype avian influenza. PMID- 12557369 TI - [Selection and characterization of peptides that specifically binding to hepatitis C virus serine protease]. AB - The HCV NS3 serine protease that plays important role in the processing of polyprotein and the replication of virus is a prime target for antiviral drugs and therapy research. Based on the crystallographic structure of HCV sreine protease, a single-chain protease was contstructed in which the central sequence of NS4A was fused to the N-terminus of NS3 serine protease domain via a flexible linker and it was expressed at high level in soluble form in E. coli. The purified protease could cleave the recombinant protein NS5ab into two parts. The purified protease was used as target to screen binding peptides from phage displayed peptide library. After three rounds of affinity screening, 37 out of 44 randomly selected phages could bind specifically with the single-chain serine protease and their specificity were verified by competitive ELISA. The 13 sequenced clones represents 6 kinds of sequences of which the amino acids composition is in bias and there is a consensus sequence: [H/F/W]-H-W-X-X-W. PMID- 12557370 TI - [Cloning and expression of Vitreoscilla hemoglobin in Streptomyces aureofaciens]. AB - Vitreoscilla hemoglobin Gene was cloned in Streptomyces aureofaciens through E. coli-Streptomyces shuttle plasmids constructed by both pIJ699-pUC19(vhb) and pIJ702-pBR322(vhb). Under low dissolved oxygen conditions, expression of hemoglobin enhanced CTC yield of engineering strain more about 40%-60% that that of control by improving oxygen transmission in cells. Hemoglobin expression by Promoter of tetracycline resistance gene was more effective for oxygen transmission than that by its native promoter regulated by dissolved oxygen concentrations in environments of locally low concentrations of dissolved oxygen. PMID- 12557371 TI - [Cloning of IL-1 beta gene and expression in E. coli]. AB - IL-1 beta cDNA was obtained by RT-PCR with the template of the total RNA extracted from leukocytes which was separated from human peripheral blood. 5' and 3' primers were synthesized according to literaturees reported sequence of IL-1 beta. IL-1 beta gene was highly expressed in E. coli and the expression level reached to about 40% of total bacteria proteins. Separation, purification and bioactivity analysis of the expressed products was performed. The purity of the final products reach more than 98%, and the culture solution of EL-4 cells induced by hIL-1 beta can promote the proliferation of CTLL-2 cells obviously. PMID- 12557373 TI - [Screening strains producing inulinase and cloning of inulinase gene]. AB - A wild-type strain AF10 producing inulinase was screened from soil samples, the strain is identified to be a A. niger. A endoinulinase gene inuA1 from A. niger AF10 was wequenced and analyzed, after PCR amplification. The results shows inuA1 is 1551 bp in length without any intron and contains 4 potential N-glycosylation sites. The conservative sequence is WMNEPN from the N-terminus. The inuA1 was cloned to pUC118 and the recombinant vector pinuA1 was obtained and transformed into E. coli JM109. The recombinant JM109/inuA1 included inuA1 was obtained. PMID- 12557372 TI - [Isolation and disruption analysis of a developmental gene of Myxococcus xanthus]. AB - FruB is an associated protein of FruA, a transcription factor essential for the development of Myxococcus xanthus. Degenerate oligonucleotide primers were designed based on the N-terminal amino acid sequence of FruB. Using genomic DNA as template, an approximate 110 bp fragment was generated and further served as a probe to screen a small genomic library of M. xanthus. A 4.5 kb SacI fragment was isolated on the basis of its homology to the probe. Disruption of fruB delayed the morphogenesis of fruiting bodies and depressed spore yield, suggesting that to some extent, FruB might be involved in the development of M. xanthus. PMID- 12557374 TI - [Analysis on integration sites of transposon-insertion mutant by transposon rescue]. AB - In order to define the chromosomal locus of the transposon integration in a mutant of Corynebacterium glutamicum, transposon rescue experiments were performed. By isolating parts of the transposon together with adjacent parts of the bacterial chromosome and subsequent sequencing of the adjacent parts, the mutant was found to have three transposon integrations, one just in front of the citrate synthase gene gltA, the other two in hitherto unknown open reading frames designated orfA and orfB. The transposon rescue experiments are proven to be a easy, practical and ideal way in analyzing the transposon integration sites. PMID- 12557375 TI - [Study on the methods for virus epidemiology in epitope level]. AB - A method for studying virus epidemiology in epitope level was established via phage random peptide library and thioredoxin surface display technique and the method was proved by test with core protein of HCV. PMID- 12557376 TI - [Study on amount and stability of cry1D and cry1Ab mRNA in Bacillus thuringiensis subps aizawai]. AB - cry1Ab, cry1C and cry1D are three typical cry1 class genes encoding protoxins in Bacillus thuringiensis subsp. aizawai strain HD-133. The expression of cry1D is obviously different from that of cry1Ab in this strain. Amount and stability of cry1D and cry1Ab mRNA in HD-133 were investigated. Northern blotting analysis showed that cry1D mRNA was more stable than cry1Ab mRNA, however, cry1D mRNA formed later 3 h and less 3.7 times than cry1Ab mRNA during the mid-phase of sporulation. Halve lives of cry1D mRNA was 18 min, but halve lives of cry1Ab mNRA was 14 min. Moreover, cry1Ab mRNA could keep stable and large amount during the post-phase of sporulation. This suggested that difference of transcriptional efficiency and initiate time might be main reason for the difference of expression of cry1Ab and cry1D. PMID- 12557377 TI - [Study on quantitative test on the DNA damage of Aspergillus flavus caused by citral with a comet analysis system]. AB - Comet assay provided a object method for DNA damage caused by physical and chemical factors. A new system of SCGE image analysis, which combines microphotodensitometry and microphotography with image analysis have been developed. Compared with the international popular, it has well at speed, convenience, exchange face of Chinese and English and print. We used it to directly determine the A. flavus' DNA damages for citral. The results show that test time was shorted 2/3 and accuracy was improved. It is probable that cell structural parameter, cell molecule and membrane could be test long, successive, movably and instantly. So the new system has a wide application. PMID- 12557378 TI - [Purification and properties of thermostable catalase in engineered E. coli]. AB - A thermostable catalase in engineered bacterium E. coli was purified to electrophoretic homogenenity by heat treatment, ammonium sulfate fractionation precipitation, DEAE-A50 ion exchange chromatography, HiPrep 16/10 Phenyl hydrophobic interaction chromatography and Superdex200 HR 10/30 size exclusion chromatography with 187.2-fold purification and 9.8% recovery. The optimum reaction temperature and pH of this recombinant catalase were 70 degrees C and 7.0 respectively. The catalase is stable below 60 degrees C and at pH range 3-8. The residual activity of the catalase was about 60% after treated at 70 degrees C for 60 minutes and 80 degrees C for 10 minutes. The apprant Km and Vmax value of the catalase were 7.75 mmol/L and 27.8 mmol.min-1.mg-1 respectively. The affects of some metal ions and compounds on this enzyme were shown. Zn2+, Ba2+, Mn2+ of 1 mmol/L could completely inactivate the enzyme, EDTA of 50 mmol/L had no affect on activity. PMID- 12557379 TI - [The cell wall-degrading activities of lysin encoded by chlorella virus PBCV-1]. AB - Lysin was isolated from Chlorella NC64A lysate which was infected by Chlorella virus PBCV-1. The enzyme property detection of lysin shows that it contains at least three enzyme activities: chitinase, chitosanase and beta-1, 3-Glucanase. This is consistent with the compose of Chlolrella cell wall. Chitinase and chitosanase, especially chitinase plays a important role in the process of virus entry. One 52 kD chitinase, one 56 kD chitosanase and one 36 kD chitosanase were obtained after purified by FPLC. PMID- 12557380 TI - [Effects of H2O2 addition on cell growth and product formation in long-chain dicarboxylic acid fermentation]. AB - When Candida tropicalis was cultivated in shakig-flask with the H2O2 addition, DCA (Dicarboxylic Acid, DCA) concentration was increased, especially at 2 mmol/L H2O2 concentration. The cytochrome P450 activity assays indicated that H2O2 addition significantly increased the activities of cytochrome P450 and DCA production positively correlated with the activities of cytochrome P450. The study on the cell growth demonstrated that the H2O2 addition inhibited the cell growth rate. However, the retarding effect was not irreversible since the cell growth rate could recover slowly to the original level after the H2O2 addition was halted. The mechanism of inducement on cytochrome P450 by H2O2 addition was also discussed in this article. PMID- 12557381 TI - [Decolorization of dyestuff and dying waste water by laccase solution with self flocculent mycelial pellets of Coriolus versicolor]. AB - Both laccase production by the white-rot fungus Coriolus versicolor and decolorization of dyestuff and dying waste water with crude solution of laccase were studied in this work. Laccase production meets the definition of secondary metabolism. For laccase production the optimum initial pH is 4.5. Addition of veratryl alcohol or elevated trace metals could both enhance the laccase activity, while Tween80 showed some inhibition. The immobilized mycelia of C. versicolor in polyurethane foam had less laccase production ability than mycelial pellets. A repeated batch cultivation process was found to be a very economical way for laccase harvest. The same pellets could be used for at least 14 times and average laccase activity of each batch could maintain 6.72 IU/mL. This method reduces the enzyme production course, medium consumption and the possibility of contamination, showing high efficient and great economic benefit. Good results were also obtained in decolorization experiments with the crude solution of laccase. With 3.3 IU/mL initial laccase activity, color removal of Acid Orange reached 98.5% after 24 h reaction. Also with 2.6 IU/mL initial laccase activity, color removal of dying waste water reached 93% after 24 h reaction. PMID- 12557382 TI - [Construction of pseudorabies virus Ea TK-/gE-/gp63- mutant strain and the study on its biological property]. AB - Using pseudorabies virus Ea strain as material, we inserted LacZ gene expression cassette into gE gene. After blue plaque and plaque purification, a recombinant virus PRVEa TK-/gE-/LacZ+ generated. Utilizing EcoR I site in LacZ gene, digested PRVEa TK-/gE-/LacZ+ genome DNA was cotransfected into PK-15 cells with plasmid pFBBS, then PRVEa TK-/gE-/gp63- generated after plaque purification. Four pairs of primers amplification demonstrated the virus was pure TK-/gE-/gp63- mutant virus. PCR product sequence indicates there were 205 bp deletion in TK gene; 1247 bp deletion in gE,gp63 and intergenic region of PRVEa TK-/gE-/gp63- mutant virus genome DNA. Inoculation to Balb/C mice with PRVEa TK-/gE-/gp63- indicates the virulence is reduced greatly. PMID- 12557383 TI - [Re-cloning of THP gene and construction of high efficient expression yector of Volvariella volvacea]. AB - PCR technique was used for amplifying THP gene in an unknown vector with primer AFP1 and AFP2. Then THP gene was ligated to pGEM T-Vector to be the plasmid pGTHP4. The plasmid pCAMBIA1301 was digested with restriction enzyme BstE II and Nco I, and digestion product was separated with 1% of agarose gel, then big fragment containing promoter was isolated and purified with the Agarose Gel DNA Extraction Kit. At the same way, the plasmid pGTHP4 was digested with restriction enzyme BstZ I and Nco I, and the small fragment containing THP gene was purified from 1% agarose gel with the Agarose Gel DNA Extraction Kit. The big fragment and the small fragment were ligated at Nco I digested cohesive-end. The ligation product was re-ligated to be cyclic plasmid by addition to a specific adapter, resulting in the pCTH823, a expression vectorof V. volvacea. PMID- 12557384 TI - [Preliminary studies on the apoptosis of Spodoptera litura cells induced by Two baculoviruses]. AB - Microscopic examination of Spodoptera litura(SL-1) cell infected with AcMNPV or SfaMNPV revealed progressive cell blebbing starting at 8 h-12 h postinfection and culminating in total cell destruction at 24 h postinfection. The fragmentation of the infected cell nuclei was observed by stained with the specific fluorescent dye DAPI. Agarose gel electrophoresis analysis of the DNA extracted from infected cells show typical DNA ladder. All these supported that both viruses infected SL 1 cells undergo apoptosis. Through lipofectin transinfection, we observed that only the DNA of AcMNPV or SfaMNPV can also induce apopotosis of SL-1 cell. Virus titer analysis revealed that neither viruses can duplicate in SL-1 cells. PMID- 12557385 TI - [Studies on cell integrity pathway and its upstream regulation factors in yeast]. PMID- 12557386 TI - [Advance of actional mechanism for the toxin of entomopathogenic fungi to insects]. PMID- 12557387 TI - [Analysis of 16S rDNA sequences and DNA-DNA hybridization of moderately halophilic bacteria from Xinjiang region]. AB - Based on the previous studies on numerical taxonomy and 16S rDNA PCR-RFLP analysis, the moderately halophilic bacteria isolated from Xinjiang Region constituted a new cluster, and the phylogenetic tree was constructed by comparing with the 16S rDNA sequences of the other moderately halophilic bacteria species. In the phylogenetic tree, most of the reference strains were clustered in a group, and the similarity values of 16S rDNA sequence were above 96%. However, AI 3, Alcanivorax borkumensis and Halobacillus litoralis were clustered in another group, and the similarity value of 16S rDNA sequences between AI-3 and Alcanivorax borkumensis was 96%, and that of 16S rDNA sequences between AI-3 and Halobacillus litoralis was 99%. The results indicated that AI-3 was different from the reference strains in phylogeny. The values of DNA homology in the new cluster were more than 70%, but the value between AI-3 and Halomonas elongata was less than 50%. Thus, the strain AI-3 possibly represent a new moderately halophilic bacteria species. PMID- 12557388 TI - [Isolation, identification and phylogenetic analysis of a thermophilic cellulolytic anaerobic bacterium]. AB - Four strains of thermophilic cellulolytic anaeobic bacteria were isolated from fresh feces, heat compost, cellulolytic mixed culture with a method based on adherence of cellulolytic bacteria to cellulose. The cells of isolates were straight or slightly curved rods that were 0.4 micron-0.6 micron x 3 microns-15 microns, Gram negative, strictly anaerobic, sulfate reduction negative, spore forming bacteria. Most of the cells had oval terminal spores, while subterminal spores, middle spores, two or more spores also could be observed and spore formation could occurred in any position. The isolates degraded cellulose filter paper, cellulose powder Whatman CF II, microcrystalline cellulose, cellulose powder MN300 and unpretreated maize stem core, sugarcane residue and rice straw. The pH and temperature ranges for growth on cellulose were 6.2-8.9 and 45 degrees C-65 degrees C respectively with the optima, 7.0-7.5 and 55 degrees C-60 degrees C, respectively. The major fermentation products from cellulose were acetic acid, ethanol, CO2, H2. The isolates could ferment cellobiose, glucose, fructose, maltose, and sorbital. The phylogenetic analysis based on 16S rDNA suggested strain EVA1 was the closest relative of Clostridium thermocellum with 99.8% sequence similarity. PMID- 12557389 TI - [Cloning and expression in Escherichia coli of oligo-1,6-glucosidase gene from Bacillus subtilis HB002]. AB - The gene coding for oligo-1,6-glucosidase of Bacillus subtilis HB002 was cloned by the shotgun-cloning method and sequenced by the chain-termination method of Sanger et al. It consists of an open reading frame of 1683 bp. The amino acid sequence of oligo-1,6-glucosidase deduced from its nuecleotide sequence predicts a protein of 561 amino acid residues with a Mr of 65.985 kD, which is 81% and 67% identical to those of oligo-1,6-glucosidase from Bacillus sp. and Bacillus coagulans, respectively, 89% and 79% similar to those of oligo-1,6-glucosidase from Bacillus sp. and Bacillus coagulans, respectively. The oligo-1,6-glucosidase gene of Bacillus subtilis HB002 was cloned into Escherichia coli expression plasmid pBV220, the result of SDS-PAGE showed that the oligo-1,6-glucosidase gene had been expressed in Escherichia coli DH5 alpha, the expressed oligo-1, 6 glucosidase has enzymatic activity. PMID- 12557390 TI - [Cloming, sequence analysis of imidase gene from Alcaligenes eatrophus and its expression in E. coli]. AB - A hydantoin-cleaving microorganism 112R4 is screened and identified to be Alcaligenes eutrophus. The resting cell of Alcaligenes eutrophus 112R4 can catalyze the hydrolysis of hydantoin, dihydropyrimidine and succinimide effectively, but not function to 5-monosubstituted hydantoins or 5,5' disubstituted hydantoins. The microorganism can utilize succinimide as a sole carbon source and nitrogen source, which indicates the presence of a complete transformation pathway of succinimide, and a hydantoin-cleaving enzyme, imidase, is suggested to be contained in this metabolic pathway. A 6 kb EcoRI-EcoRI fragment isolated from the genome DNA of Alcaligenes eutrophus 112R4 is shown to be correlative with the transformation of succinimide. A 2 kb DNA fragment containing the gene of imidase is subcloned and sequenced. Deletion analysis verifies that one open reading frame of 876 nucleotides, which encodes a peptide of 291 amino acids, with a calculated molecular weight of 33688, is responsible for the encoding of imidase. This is the first report of the nucleotide and amino acid sequences of imidase (GenBank accession number: AF373287). A homology search performed in protein database reveals an identity of 14% with polysaccharide deacetylase conserved domain, an identity of 60% with N-terminal 20 amino acids of Blastobacter sp. A17p-4, but no apparent similarity with all known cyclic amide-cleaving enzymes. This result suggested that the imidase should be classified as a new member of cyclic amidases. Under the control of lac promoter and IPTG induction, the imidase activity of transformed E. coli reached 3200 U/L, which is about 7-fold higher than that of gene donor strain. PMID- 12557391 TI - [Cloning and expression of a osmoregulatory gene pro B from halotolerant Bacillus subtilis]. AB - A 1.3 kb fragment is cloned from halotolerated Bacillus subtilis 93151 with PCR amplification method, and its positively-directionally inserted fragment can complemented with proB-E. coli by function test. Halotolerated ability of E. coli DH5 alpha having this recombination plasmid rises from 2% to 4% in minimal medium. The nucleotide sequence of this fragment is obtained by primer walking method. Nucleotide sequence of this fragment 167-1269 bp translates a protein which has 370 amino acid by sequence analysis through DNAsis program. There are non-typical-10 sequences, typical-35 sequences and a Ribosome binding site of this fragment in its upstream sequence, and there is flanking sequence, which has best efficiency of beginning translating. Homologues comparision, of nucleotide and amino acid sequences of this fragment and those of gene in gene bank shows that homogenous of Nucleotide sequences and amino acid sequences of this fragement and Bacillus subtilis 168 are respectively 81%, 90%, which prove that this gene is certainly a pro B gene. This protein translated by this fragment has several absoluter conservative domain which have been correlating closely with forming active center of enzyme and tri-dimension structure of active center, compared amino acid sequences of this fragment and pro B genes of thirty kinds of different microorganism. PMID- 12557392 TI - [Characterization of cry gene and broad spectrum against lepidopteran of Bacillus thuringiensis subsp. colmeri 15A3]. AB - Bacillus thuringiensis wild type strain 15A3 belongs to subspecies colmeri serotype H-21. RFLP and PCR analysis show that it contains six types of ICP genes: cry1Aa, cry1Ac, cry1Ca, cry1D, cry1I and cry. The sequence of the 1.45 kb N-terminal fragment of cry1Aa differed from that of published. SDS-PAGE showed that the crystal consists of proteins with molecular weight about 130, 79, 70, 65, 51 and 45 kD. Strain 15A3 didn't sysnthesize heat-stable beta-exotoxins according to test of house fly aberration. The 1.2 tons fermentative production exhibited high toxicity against three lepidopteran pests: H. armigera, S. exigua and H. cunea. It was proved that wild type strain can produce a broad specturm of ICP. PMID- 12557393 TI - [Analysis of function of the 452 bp sequence proceeding the pgiB gene in Bacillus stearothermophilus]. AB - By random mutagenesis with chemical mutagen N-methyl-N'-nitro-N-nitrosoguanidine, a shuttle promoter probe vetor mutant pPGV5 (tr65) was obtained. A single site mutation from G to T was discovered at nucleotide + 238 of kan gene encoding kanamycin nucleotidyltransferae by sequence analysis. Using the gene pheB that encodes thermostable catechol 2'3-dioxygenase from Bacillus stearothermophilus FDTP-3 as the reporter gene, a transcriptional fusion plasmid pPGVPB452 was constructed and electroportated into B. stearothermophilus. By analyzing the reporter protein, it proved that the 452 bp sequence proceeding the coding region of phosphoglucoisomerase pgiB gene from B. stearothermophilus T521, which contained a putative promoter by sequence analysis, but has no transcriptional activity in B. stearothermophilus. PMID- 12557394 TI - [Construction of DNA transfer system of Streptomyces tenebrarius]. AB - To establish a gene transfer ssystem in Streptomyces tenebrarius, several methods including PEG-mediated transformation of protoplasts, conjugal transfer were investigated. Many attempts were made to introduce plasmid pIJ702 into Sreptomyces tenebrarius. It was found that plasmid pIJ702 isolated from S. lividans TK24 failed to transform the protoplasts of Sreptomyces tenebrarius. No transformant was achieved even if the protoplast was inactivated by heat treatment or dsDNA was converted ssDNA before transformation. All the results suggested that Sreptomyces tenebrarius exists a strong restriction and modification system for the transformation of foreign DNA. A recombinant E. coli ET12567 (pUZ8002, pHZ132) was obtained by transforming E. coli ET12567(pUZ8002) with oriT-containing E. coli-Sreptomyces shuttle plasmid pHZ132. In mating experiments, E. coli ET12567 (pUZ8002, pHZ132) was the donor, and the recipient was Sreptomyces tenebrarius 9904 spores after pregerminating by heat shock. Matings between dornor and recipient were conducted. Plasmid pHZ132 was introduced into Sreptomyces tenebrarius 9904 by conjugation from E. coli ET12567. The transfer system of Sreptomyces tenebrariu was established by conjugation. S. tenebrarius 9904 protoplasts were transformed by plasmid DNA modified by the host itself, and the transformation frequency was about 10(3)/microgram DNA (pHZ132). PMID- 12557395 TI - [Analysis the influence of palindrome structure to gene expression by constructing combination system]. AB - Palindrome sequence is very common in DNA, it usually acts as cis-elements of many regulative factors. In this paper, we design two palindrome sequences which is different in length, construct Cre-loxp combination system, then co-transform two kinds of plasmids, one contains cre gene, the other contains loxp site to E. coli DH5a strain. We judge the expression degree of cre gene in E. coli by examining the changes of plasmid maps of co-transformed strains, then conclude that 20 bp or shorter than 20 bp palindrome sequence uninhibit the expression of downstream genes, however, 34 bp or longer than 34 bp palindrome sequence can inhibit that. PMID- 12557396 TI - [Analysis of POX4 and POX5 gene encoded proteins of Candida tropicalis 1230]. AB - POX4 and POX5 were cloned from Candida tropicalis with a simple method. The sequencing results show differences of POX5 between strain 120 and strain pk233. The searching of these two sequences against the curreent collection of Pfam profile HMMS located regions that belong to known domain families. Secondary structure prediction shows that N-terminal secondary structure of PXP4 and PXP5 are homologous to that of pig liver medium-chain acyl-coA dehydrogenase. Thus, the N-terminal of PXP4 and PXP5 may be involved in FAD binding. PMID- 12557397 TI - [cDNA cloning and sequence analysis of genome segment S7 of rice black-streaked dwarf virus]. AB - Genome segments 7 of zhejiang and Hebei isolates of rice black-streaked dwarf virus (RBSDV) were amplified and sequenced. Segment 7 of Zhejiang isolate was consisted of 2193 nts (EMBL accession no. AJ297427) in length and that of Hebei isolate was 2190 nts (AJ297428). Both segments contained two non-overlapping open reading frames (ORFs), which encoded two polypeptides with molecular weights of 41 kD and 36 kD. These two segments shared 99% nucleotide identity, 100% and 94.4% amino acid identities of ORF1 and ORF2, shared 93.5% and 93.8% identities at nucleotide level, 98.1% (ORF1) and 96.5%/97.8% (ORF2) at amino acid level with S7 of Japanese RBSDV, and shared 85.1% and 85.3% identities at nucleotide level, 92.3% (ORF1), 85.5%/86.8% (ORF2) at amino acid level with S6 of Italian MRDV. PMID- 12557398 TI - [Sequence analysis of the fusion (F) protein genes of four Newcastle disease virus strains causing clinical disease in geese]. AB - Four strains of Newcastle disease virus(NDV), which have proven to be responsible for the disease outbreaks in south and east China regions called "geese paramyxovirus infection" or "avian paramyxovirus infection of geese", were examined for the features of their fusion(F) protein genes. A 1700 nucleotides(nt) fragment of the F gene (1705 nt) of each of the 4 strains were sequenced, the results revealed that the NDV strains of goose origin share common transcriptional sequence and translational start position with the hitherto reported NDV, the F gene homologies among the 4 strains were 97.8%-98.9%, while the corresponding homologies between these strains and the standard challenge strain F48E8 were only 86.0-86.8%. The deduced amino acid sequences near cleavage site of F0 proteins showed a 112RRQKR/F117 motif, comforming well with that of the virulent strains. The cleavage sites distribution of 3 restriction enzymes(Hinf I, Basto I, Rsa I) in 75% region of the F gene(between 334-1682 nt), which is used to group NDV strains into different genotypes, were also analysed with the aid of computer program MegAlign, it is very interesting to find that the 4 strains showed a distinct distribution pattern from all the genotypes so far reported(genotype I-VIII), characterized by the simultaneously presence of a Bsto I site at 752 nt and two Rsa I sites at 872 nt and 973 nt. Additionally, 3 out of the 4 strains, despite their different geographical locations, showed a novel Rsa I site at position 1249 which is absent in all of the other genotypes. The 4 NDV strains of goose origin were essentially virulent ones, and probably belong to a new genotype. PMID- 12557399 TI - [Studies on hydantoinase producing conditions of Arthrobacter K1108]. AB - The hydantoinase-producing conditions from strain Arthrobacter K1108 were investigated. It is shown that the hydantoinase in the strain is intracellular and inducible. Its optimal inducer is 5-benzylhydantoin, while 5 indolylmethylhydantoin and 5-phenylhydantoin cannot induce the production of hydantoinase in K1108. A gratuitous inducer was designed, with which the hydantoinase production is 343% as much as that with 5-benzylhydantoin. The media for culturing the bacteria were screened and optimized. Under optimal conditions, the specific activity of K1108 cells reached 10.8 U/mL. PMID- 12557400 TI - [A possible role of a low-molecular-weight peptide from Gloeophyllum trabeum in cellulose degradation]. AB - A low-molecular-weight peptide(named Gt factor) was first isolated and purified from the extracellular culture of a brown-rot fungi Gloeophyllum trabeum. It could produce hydroxyl radical HO. in presence of O2 and Fe3+, might destroy hydrogen bonds of cellulose by HO.-involved oxidative mechanism, which was quite different from hydrolysis mechanism of cellulase in filamentous fungi. Gt factor might attack crystalline region of cellulose, bring about more reducing ends and non-reducing ends exposed, thus making cellulose accessible to further degradation. PMID- 12557402 TI - [The optimization of guanosine fermentation based on process parameter correlation analysis]. AB - The characteristic of Bacillus subtilis fermentation process of guanosine on 50 L fermentor was analyzed. Based on determination of on-line and off-line parameter, using correlation analysis, the technology study of physiologic regulation was combined with the metabolic flux distribution of synthesis process. The metabolic flux shift from HMP to EMP and TCA cycle during fermentation was found. The reason of the flux shift was preliminary analyzed, based on which the procedure was optimized to increase the yield of guanosine to 30 g/L. PMID- 12557401 TI - [Study on biochemical mechanism of citral damage to the A. flavasi's mitochondria]. AB - Adopted biochemical method and combinded with Scanning Electron Microscope(SEM) to observe, study the inhibition-growth mechanism to A. flavus when citral was permeated to the cell and damaged mitochondria, the result suggested that the mitochondria changed in many kinds of unrule shape and the oxidation-reduction system was destroyed, when the citral reached at the level of anaphylactic consistency to A. flavus. Compared with the control group, activities of MDH and SDH in experimental group non-reversibly decreased 27.1% and 23.8% respectively, and grandully lost. When respectively used the succinate, pyruvate and alpha ketoglutarate as substrates, the respiration speed of mitochondria separatelly decreased 24.1%, 36.1%, 14.3%. It suggested that the citral could inhibit the biosynthesis of the mycelial DNA, RNA, lipoid and protein, so promoted to dath. PMID- 12557403 TI - [The solubilization of four insoluble phosphates by some microorganisms]. AB - Four insoluble phosphates of ferric phosphate (Fe-P), aluminum phosphate (Al-P), fluorapatite (FAP) and rock phosphate (RP) were used as a sole phosphorus resource for some phosphate-solubilizing microorganisms. It was found that there was significant difference in solubilizing these phosphates by the tested isolates. The fungi normally were more powerful than the bacteria in dissolving the phsophates. The microorganisms generally solubilized more phosphate when supplied with NO3- than with NH4+. However, the isolates of 2TCiF2 and 4TCiF6 had much higher capacity to solubilize FAP and Al-P respectively in NH4+ medium. Most of the isolates solubilized readily FAP and RP, and then Al-P. Ferric phosphate was the least soluble to these isolates. Only isolate 2TCiF2 showed strong ability to solubilize Fe-P. In particular, two Aspergillus sp. had much higher capacity of dissolving Fe-P when suppled with NO3-. The isolates of Evwinia sp. 4TCRi22 and Enterobacter sp. 1TCRi15 had higher capacity of solubilizing FAP. But two Arthrobacter sp. showed the highest activity in RP medium. It is supposed that complexion of organic acids with metals may be the main reason for these isolates to solubilize the phosphates. However, other chelant substances may be much more important for Enterobacter sp. and Erwinia sp. to release phosphorus from the phsphates. PMID- 12557404 TI - [Sequencing and analysis of flocculation gene(FLO1G)]. AB - The sequence of the flocculation gene (FLO1G) was determined. The result of sequcencing showed that: the cloned gene contains a large open reading frame (ORF) of 3936 bp and encodes for a protein of 1312 amino acid. According to the result of homologous analysis, the cloned gene is homologous to FLO1 but with 675 bp deletion in the ORF region. The missing part belongs to one of the four repeated sequence family of FLO1. Since the cloned DNA fragment can trigger strong flocculence to non-flocculent strain S. cerevisiae YS58, we concluded that the missing part is not the crutical part for the flocculent ability of the gene. PMID- 12557405 TI - [The resistance conferred by the R/M system LlaBIII against bacteriophages]. AB - LlaBIII, isolated from the naturally occurring plasmid pJW566 from Lactococcus lactis subsp. cremoris W56, encoded a restriction and modification (R/M) system. The plasmid pJK1 carrying the R/M system LlaBIII was transformed into L. lactis IL1403 with type I R/M system located on chromosome and the strain MG1614[pAW601] with AbiS gene on plasmid pAW601, respectively. The transformants obtained showed stacking resistance against bacteriophages. The plasmid pJK1 was transformed into industrial strains L. lactis SMQ86 and CHCC2281, the transformants showed the EOP of the bacteriophages decreased by 10(-3) and 10(-5), respectively. The results indicated that the R/M system LlaBIII could protect strains from bacteriophages in dairy fermentation. PMID- 12557406 TI - [Progress in microbial proteomics]. PMID- 12557407 TI - [Programmed cell death in bacteria]. PMID- 12557408 TI - [Characteristics and application of thermophilic enzymes]. PMID- 12557409 TI - Anesthesiologists and perioperative hemostasis: should we feel concerned? PMID- 12557410 TI - Two new antithrombotic agents (fondaparinux and ximelagatran) and their implications in anesthesia. AB - PURPOSE: To describe two new antithrombotic agents (fondaparinux and ximelagatran), their mechanisms of action, the clinical studies available and some implications (regional anesthesia, monitoring and antidote) in anesthesia. SOURCE: Recently published articles on these two new drugs were reviewed. PRINCIPAL FINDINGS: These two drugs have several interesting properties. Both are of non animal origin, do not induce thrombocytopenia and do not require laboratory controls. In recent studies fondaparinux has shown a better efficacy than low molecular weight heparin in major orthopedic interventions. Ximelagatran, which can be given orally and is in phase III investigation, could replace vitamin K antagonist in the future. CONCLUSION: Anesthesiologists should be aware of these two new agents, one (fondaparinux) being now already registered in some countries. PMID- 12557411 TI - Reversing anticoagulants both old and new. AB - PURPOSE: Reversal of pharmacologic anticoagulation is an issue that arises when an anticoagulated patient has major bleeding or when a patient on chronic anticoagulant therapy requires urgent reversal of anticoagulation, for example, for surgery. SOURCE: We reviewed the literature to determine what strategies are available to reverse anticoagulation caused by older agents, such as warfarin or unfractionated heparin (UFH), as well as newer agents, for example, low-molecular weight heparin, danaparoid, fondaparinux, lepirudin, and argatroban. PRINCIPAL FINDINGS: Specific "antidotes" exist for the "classic" anticoagulant agents: protamine sulfate for UFH, and vitamin K for warfarin. However, vitamin K only begins to reverse warfarin's anticoagulant effect by four to six hours, so urgent situations additionally require blood products, such as plasma (fresh frozen or cryosupermatant plasma), prothrombin complex concentrates, or, possibly, recombinant factor VIIa. A growing problem arises from the increasing use of new anticoagulants that lack specific antidotes. For example, protamine sulfate reverses only about 60% of the anti-factor Xa activity of low-molecular-weight heparin, has negligible effects on danaparoid (a mixture of anticoagulant glycosaminoglycans used to treat heparin-induced thrombocytopenia) and fondaparinux (a novel synthetic antithrombin-binding pentasaccharide with exclusive anti-factor Xa activity approved in the United States for antithrombotic prophylaxis following orthopedic surgery). The new direct thrombin inhibitors (e.g., lepirudin, bivalirudin, argatroban) also have no specific antidote. CONCLUSION: Newer anticoagulant agents generally lack specific antidotes. Thus, careful choice of an anticoagulant agent and laboratory monitoring where appropriate are needed to minimize risk of bleeding complications. PMID- 12557412 TI - Antiplatelet agents in the perioperative period: expert recommendations of the French Society of Anesthesiology and Intensive Care (SFAR) 2001--summary statement. AB - PURPOSE: Antiplatelet agents are administered to an increasing number of patients. Preoperative treatment with these agents represents a major problem for the anesthesiologist. The results of a French expert meeting on their perioperative management are reported. METHODS: Responses to questions formulated by the Organizing Committee were drafted by a group of experts and reviewed by a multidisciplinary. Reading Committee. Recommendations were classified (grade) according to the evidence level of the studies supporting them. PRINCIPAL FINDINGS: First, antiplatelet agents have a variable effect on hemostasis as far as bleeding risk is concerned. Aspirin and non-steroidal anti-inflammatory drugs (NSAIDs) increase intra- and postoperative bleeding moderately, but not transfusion requirements. Very few data are available on clopidogrel and ticlopidin. Anti-glycoprotein (GP) IIb/IIIa agents may increase bleeding when surgery is required in proximity with their administration. Second, the common practice of withdrawing antiplatelet agents is now challenged because an increased incidence of myocardial infarction has been reported in patients in whom treatment was interrupted. Third, aspirin should not be withdrawn for most vascular procedures and in several additional settings. When a definite increase in intraoperative bleeding is feared, or when surgical hemostasis is difficult, aspirin, clopidogrel or ticlopidine can be replaced by short-acting NSAIDS, given for a ten-day period and interrupted the day before surgery. Platelet transfusion should only be given when overt bleeding is observed. Postoperatively, antiplatelet treatment should be resumed immediately after surgery (first six hours). CONCLUSION: Anesthesiologists should be aware of the indications, potential complications and means of substitution of these agents. PMID- 12557413 TI - Heparin-induced thrombocytopenia and the anesthesiologist. AB - PURPOSE: All physicians who use heparin should be aware of immune heparin-induced thrombocytopenia (HIT), including anesthesiologists who may need to provide intraoperative anticoagulation for a patient who urgently requires cardiac or vascular surgery but who has acute HIT or a history of recent HIT. SOURCE: The literature dealing with HIT of relevance to anesthesiologists was reviewed, including studies of HIT antibody formation following intraoperative use of heparin; acute respiratory or cardiac arrest following i.v. bolus heparin indicating rapid-onset HIT; acute thrombocytopenia and thrombosis complicating intraoperative heparin use; circumstances in which it might be acceptable to administer heparin despite a previous history of immune HIT; and alternative anticoagulant approaches that can be used to manage cardiac or vascular surgery in a patient with acute or recent HIT. PRINCIPAL FINDINGS: Intraoperative exposure to heparin can trigger formation of HIT antibodies, and occasionally even lead to "delayed-onset" HIT. Acute respiratory or cardiac arrest following i.v. bolus heparin, or the abrupt occurrence of intraoperative "white clots," suggests a diagnosis of rapid-onset HIT, particularly if the patient recently received heparin. Several approaches are available to manage cardiac or vascular surgery in a patient with acute or recent HIT, so the treatment chosen depends upon local experience and monitoring capabilities. Several months after acute HIT, and particularly when HIT antibodies are no longer detectable, it may be acceptable to use heparin for intraoperative anticoagulation. CONCLUSION: HIT is an infrequent but important topic for anesthesiologists because of the urgency and complexity of the various associated management issues. PMID- 12557414 TI - [Localization of non-monoaminergic aromatic L-amino acid decarboxylase neurons (D neurons) in the human striatum and their functional significance]. AB - It has recently been reported that the human corpus striatum, especially its ventral part, named as the nucleus accumbens, contains numerous non-monoaminergic aromatic L-amino acid decarboxylase (AADC; the second-step monoamine synthesizing enzyme) neurons (D-neurons). D-neurons are the neurons immunoreactive for AADC but not immunoreactive for dopamine or serotonin. They lack the first-step monoamine synthesizing enzymes, tyrosine hydroxylase and tryptophan hydroxylase. AADC is also the rate-limiting enzyme of phenylethylamine (PEA) synthesis. D neurons might participate in the manifestation of efficacy of pharmacotherapy for Parkinson's disease by uptaking monoamine precursors including L-dopa or droxidopa (L-threo-DOPS) and by converting them to dopamine or noradrenaline, respectively. As the nucleus accumbens is one of the brain regions that are involved in the pathogenesis of schizophrenia and drug dependence, D-neurons might be related to the etiology of these mental disorders. It has also been suggested that striatal D-neurons are the pluripotential cells that have compensating functions against aging or degeneration. PMID- 12557415 TI - [Survey on the attitudes of co-medical students at Chiba University toward observation of dissection of human bodies]. PMID- 12557416 TI - Birth, growth and elimination of a single synapse. AB - Synapses are functional units regulating information flows in the neuronal circuits. How synaptic junctions are formed and remodelled is a fundamental question in developmental neurobiology. In recent years, it has become possible to visualize the formation, maintenance and remodelling of a single synapse by using new imaging methods. These studies, identifying synaptic structures by lipophilic dye markers and genetically modified synaptic molecules with fluorescent proteins, provided new insights into synapse development and maturation. Experimental evidence indicates very rapid assembly of both presynaptic and postsynaptic marker proteins at newly formed synaptic junctions. Morphological expansion of the synaptic junctional membrane is tightly coupled to both efficacy of the presynaptic neurotransmitter release and postsynaptic receptor distribution. The elimination process of pre-existing synapses has also been reported, and evidence for persistent remodelling of synaptic junctions has been provided. Information regarding birth, maturation and elimination of a single synapse is accumulating and will influence our concepts about how neuronal circuits are organized and maintained. PMID- 12557417 TI - Human liver caudate lobe and liver segment. AB - Recently, the caudate lobe has seemed to be the final target for aggressive cancer surgery of the liver. This lobe has five surfaces: the dorsal, left and hilar-free surfaces and the right and ventral-border planes. Surgeons have divided the caudate lobe into three parts: Spiegel's lobe, which is called the 'caudate lobe and papillary process' by anatomists, the caudate process, viewed as almost the same entity by anatomists, and the paracaval portion corresponding to the dorsally located parenchyma in front of the inferior vena cava. All three parts are supplied by primary branches originating from the left and right portal veins, including the hilar bifurcation area. The hilar bifurcation branch often (50%) supplies the paracaval portion and it sometimes (29%) extends its territory to Spiegel's lobe. It was postulated by Couinaud that the paracaval portion or the S9 is not defined by its supplying portal vein branch but by its 'dorsal location' in the liver. Couinaud's caudate lobe or dorsal-liver concept cause, and still now causes, great logical confusion for surgeons. We attempt here to describe the margins of the lobe, border branches of the portal vein, the left/right territorial border of the portal vein or Cantile's line and other topics closely relating to the surgery within these contexts. Finally, the caudate lobe as a liver segment will be discussed. PMID- 12557418 TI - Biological action of parathyroid hormone (PTH)-related peptide (PTHrP) mediated either by the PTH/PTHrP receptor or the nucleolar translocation in chondrocytes. AB - Parathyroid hormone (PTH)-related peptide (PTHrP) has been believed to act by binding the common receptor to PTH (PTH/PTHrP receptor). However, PTHrP is localized not only in the secretory pathway, but also in nucleoli by virtue of its nucleolar targeting signal (NTS). This review demonstrates the bipartite action of PTHrP on chondrocytes, the receptor-mediated and -independent signaling pathway. Mice with deletion of the PTHrP gene were characterized by a chondrodysplasia due to markedly reduced proliferation of epiphyseal chondrocytes. The PTH/PTHrP receptor was localized mainly in proliferative chondrocytes in the epiphyseal cartilage, indicating that PTHrP modulates normal proliferation via the receptor. In contrast to the receptor-mediated action, the mid-region of the amino acid sequence of PTHrP contains an NTS. The PTHrP translation was found to initiate from both methionine-coding AUG and downstream leucine-coding CUGs in its signal sequence. When translated from CUGs, PTHrP accumulated in the nucleoli, and the translation from AUG localized PTHrP in both the Golgi apparatus and nucleoli. Therefore, nucleolar PTHrP appears to be derived from the translation initiating from both AUG and CUGs. A chondrocytic cell line expressing a full-length PTHrP, but not PTHrP lacking NTS, were resistant to apoptosis caused by serum depletion, suggesting that the nucleolar PTHrP in chondrocytes serves as a survival factor against apoptosis. Thus, PTHrP regulates chondrocyte proliferation, differentiation and apoptosis by mediating its receptor or acting directly on the nucleolus. PMID- 12557419 TI - Phenotypic switching of in vitro mandibular condylar cartilage during matrix mineralization. AB - In order to analyze the phenotypic conversion of chondrocytes, mandibular condyles of mice and rabbits were cultured under cell and organ culture systems, and then examined by a combination of morphological and biochemical procedures. In organ culture, mandibular condylar cartilage (MCC) obtained from newborn mice began to mineralize from the central zone and then progressively widened towards the peripheral zone. Electron microscopic observations showed that with the increasing duration of the organ culture, chondrocytes at the central zone converted into spindle-shaped osteoblastic cells accompanying the formation of the bone type of thick-banded collagen fibrils. To obtain a better understanding of the chondrocytic conversion, immunolocalizations for type I and type X collagens and osteocalcin (OC) were examined in mouse MCC cells in cell culture. Type X collagen and OC were expressed almost simultaneously at the late stage of culture, and type I collagen was detected along the calcified nodules after the production of these proteins. Northern blot analysis in cell cultures of rabbit MCC indicated that type II collagen and alkaline phosphatase (ALPase) messenger ribonucleic acids (mRNAs) were highly expressed at day 7, but subsequently decreased. In contrast, mRNA for type I collagen was expressed at a low level on day 7 and peaked on day 12. The present results suggest that, morphologically and biochemically, cellular modification in MCC cells under culture conditions occurs at a cellular morphological level and also at marker-gene-expression level. PMID- 12557420 TI - Examination of the variations of lateral femoral cutaneous nerves: report of two cases. AB - The origins, courses and relations of lateral femoral cutaneous nerves (LFCNs) were examined bilaterally in 28 cadavers, and the variations were observed in two. On the right side of one cadaver, the ventral rami of the first and second lumbar spinal nerves were united and then this nerve was divided into four branches. From medial to lateral, these branches were the obturator nerve, the femoral nerve, the medially located LFCN and the laterally located LFCN. On the left side of another cadaver, there were three LFCNs. All of these nerves pierced the psoas major muscle anterolaterally. Two of these nerves, which pierced the psoas major muscle more proximally than the third, united with each other by a communicating branch anterior to the iliacus muscle. These types of variations are very important, especially in the presence of paresthesias or pain in the anterior thigh, lateral thigh and gluteal region. In these cases, surgeons must always remember the possible variations of the LFCN during surgical procedures in order to prevent injury and the occurrence of meralgia paresthetica. PMID- 12557421 TI - Distinct intergender difference in the femoral artery ramification patterns found in the Turkish population: angiographic study. AB - Using 600 bilateral femoral angiographies (300 male lower extremities and 300 female lower extremities), we observed configurations of the femoral origins of the medial circumflex femoral artery (MCFA), lateral circumflex femoral artery (LCFA) and the deep femoral artery. The distinct intergender difference was found in the incidence of direct origin of the MCFA from the femoral artery (17.8% and 80.0% in male and female limbs, respectively). The LCFA also exhibited a marked difference between genders in the incidence of direct origin. The Turkish population seemed to be characterized by an intergender difference in specific somatic arterial morphology. PMID- 12557422 TI - The clinical anatomy of coronary arteries. AB - In view of the fact that, in cardiology and coronary surgery, peculiarities and anomalies of the coronary arteries pose a great potential risk for myocardial ischemia resulting in arrhythmia, angina, infarction, and sudden death, a survey of the morphology of the coronary arteries in cadaver specimens (n = 200) and corrosion casts (n = 30) was undertaken. The results are not divided into those obtained from subjects with or without coronary heart disease. The randomly selected material from two anatomical institutes, of the Universities of Munich and Wurzburg, was considered representative of an average group of septuagenarians and octogenarians in the German population. It is the patient of this age that the young medical students of today (the future physicians) will be confronted with and called upon to treat. The study begins with a review of the nomenclature and a consideration of unofficial (but often clinically used) terms and synonyms for coronary arteries and their branches, followed by a description of modern diagnostic techniques based on the presence of normal and anomalous cardiac vessels. It continues with a demonstration of "normal" macro- and microanatomy, a description of the most frequent ramification pattern and as well of the many peculiarities of the coronary arteries. These are briefly enumerated here: ectopic coronary ostia and aberrant and accessory arteries; angles of origin and division; three-dimensional orientation of the coronary stems; intramural, intraluminal, and intracavitary courses; avascular coronary and interventricular sulci; extracardiac noncoronary myocardial supply; atrial branches, sinuatrial node artery, atrioventricular node artery, principles of septal arterial supply, variations of the right and left superior septal arteries; endomural, intracoronary, intercoronary, and noncoronary anastomoses and collaterals; measurement and evaluation of age-related and atheromatous coronary calcifications; multiple coronary aneurysmata. PMID- 12557423 TI - [Arterial hypertension in the cat. A pathobiologic and clinical review with emphasis on the ophthalmologic aspects]. AB - Arterial hypertension in cats appears to be an often underdiagnosed problem. Sudden disturbances of vision caused by intraocular haemorrhage and/or detached retina are often related to hypertension. The ability to measure blood pressure routinely in cats, by using an indirect method, has increased knowledge of feline hypertension in recent years. In cats mainly secondary hypertension is described, caused by as a consequence of renal disease, hyperthyroidism, chronic anaemia, primary aldosteronism, and a high-salt diet. This article describes the (patho) physiology of blood pressure control, the different methods of blood pressure measurements and the causes, clinical manifestations, and possibilities of antihypertensive therapy. Given our current knowledge, blood pressure should be measured regularly in older cats (> 10 years), especially in those with renal insufficiency, hyperthyroidism, or visual disturbances of unknown origin. Blood pressure measurements using the Doppler method is a relatively cheaply, quick and simple, method with enough reliability. Hence this method should be incorporated in veterinary practice. PMID- 12557424 TI - [Recognized veterinarians soon to be accepted by the government?]. PMID- 12557425 TI - [A flying terrier]. PMID- 12557426 TI - [Integrity]. PMID- 12557427 TI - [A congress with a full future. An impression of the Group Groot Dagen 2002]. PMID- 12557428 TI - [Everything under control? Recognition of professional burnout problems in veterinarians]. PMID- 12557429 TI - [Radionuclides for metastatic bone pain palliation]. AB - The paper overviews the role of systemic radionuclide therapy in patients with disseminated bone metastases. Most patients with bone metastases experience painful symptoms. Systemic radioisotope therapy is an alternative to traditional hemibody radiation in cases of multiple, diffuse metastases. Usually given as a single i.v. slow infusion it provides a pain relief beginning in one to three weeks, with a mean duration up to several months, depending on the kind of radioisotope applied. The paper overviews the role of unsealed source therapy with these bone-seeking radiopharmaceuticals in palliating pain, improving quality of life, indications, contraindications and complications of this therapy are discussed, as well as cost-benefit aspects. PMID- 12557430 TI - [Preliminary evaluation of microsatellite instability in skin cancers]. AB - Microsatellites of DNA--short, tandem repeat segments dispersed throughout the human genome--probably play a role in carcinogenesis. In this study the presence of microsatellite instability (MSI) was evaluated in skin cancers. Only one specimen (basal cell carcinoma) out of twelve skin cancers exhibited MSI at chromosome 2. MSI was not found in benign skin lesions (seborrhoeic verruca, keratoacanthoma, senile keratosis). PMID- 12557431 TI - [Arginase as a marker of cancerogenesis. I. Monitoring patients after resection of colorectal cancer]. AB - Arginase activity in blood serum of 40 patients with primary colorectal cancer determined one day before tumour resection, was almost 5-times higher than that in serum of healthy blood donors. Preoperative increase of arginase activity was observed in 28 out of 40 patients (70%). Six days after tumour resection, arginase activity decreased to the normal value in most studied patients. In 32 patients (86%), the activity remained low during three years of surveillance, and no symptoms of carcinogenesis were observed. However, the postoperative raise of arginase activity was demonstrated in 5 patients--in 3 of them colorectal cancer recurred and 2 developed liver metastases. The increase of arginase activity in blood serum, after resection of primary colorectal cancer, may indicate tumour recurrence or liver metastases. Therefore, monitoring of postoperative arginase activity in patient's serum may be of diagnostic value. PMID- 12557432 TI - [Arginase a marker of cancerogenesis. II. Monitoring of patients after resection of colorectal liver metastases]. AB - Preoperative activity of arginase in blood serum of patients with colorectal cancer metastases to the liver is much higher than in serum of healthy blood donors. Before tumour resection in serum of 100 patients two cut-off levels of arginase activity were observed--in 65 subjects the activity ranged from 10 to 70 U/(group I), and in 35 subjects (group II), from 100 to 200 U/l. The raised arginase activity was observed in 83% of patients from group I, and in all studied patients (100%) from subgroup II. After liver surgery, arginase activity decreased to the normal value in the majority of patients. During three years of surveillance, in 88% and 63% of patients from either subgroup, respectively, no significant increase in arginase activity was observed, and the patients remained cancer-free. However, the rise of arginase activity was demonstrated in 13% and 37% subjects from either subgroup, respectively--the patients developed second liver metastases or died. Metastases and deaths were observed more often in patients from group II than I. The rise in arginase activity in blood serum after resection of colorectal cancer metastases to the liver indicates the possibility of new cancerogenesis. The special group at risk are patients with high (100 or more U/l) preoperative serum arginase activity. PMID- 12557433 TI - [Significance of carcinoembryonic antigen levels in peritoneal washings in clinical analysis of patients with colorectal cancer]. AB - The aim of this study was to analyse the level of CEA in peritoneal washings (CEAp) in patients with colorectal cancer (crc) in comparison to the serum CEA level. The study involved 87 patients with crc, the control group included 13 patients operated on due to vascular diseases. At the time of laparotomy, after administration of 200 ml saline, peritoneal washings were collected from the peritoneal cavity. After concentrating, the CEAp level was determined using an immunochemiluminescence assay kit. The cutoff value was set at 100 ng/g protein. The data were analysed using Student t-test, Welch test and Mann-Whitney test. The mean CEA levels in peritoneal washings in patients with colorectal cancer were statistically higher (p < 0.001) than in the control group (except stage A). There was no correlation between CEAp levels and sex, histological type and differentiation, Duke's stage (except D2). The mean CEAp levels were significantly higher in colonic tumour, than in cancer in rectal location (P < 0.03). Positive correlation between CEA level in peritoneal washings and serum was found. PMID- 12557434 TI - [Tumor markers TPA and Cyfra 21.1 in patients with non-small cell lung cancer after surgery and chemotherapy]. AB - In 27 patients with operable non-small cell lung cancer (NSCLC) submitted to radical surgery followed by 3 cycles of chemotherapy (cht) serum concentrations of Cyfra 21.1 and TPA were studied. The measurements were performed before and 14 days after surgery, before each cht and every 60th day after cht was completed, for 2 years. Seven patients died during the follow up. There was no significant correlation between preoperative cyfra 21.1 and TPA serum concentrations and stage of diseases or histologic types of NSCLC. Initial concentrations of the two markers had no prognostic meaning. A significant decrease of 2 markers was observed after surgery in the whole group and in patients with therapy success. While adjuvant cht did not influence significantly serum concentrations of the markers, we showed a significant elevation of 2 markers about 4 months before death. It seems that establishing of values of Cyfra 21.1 and TPA in the patient's follow up may be useful in recognition of tumour relapse. PMID- 12557436 TI - [Value of interleukin-8 determination in diagnosis of benign and malignant breast tumor]. AB - The studies were performed in women with breast carcinoma, benign breast tumour and in a control group. Serum levels of IL-8 were determined 1-2 days before surgical procedure, according to the enzyme-linked immunosorbent assay (ELISA). Pretreatment levels of IL-8 were significantly increased in carcinoma patients in relation to the benign tumour and the control group. The frequency of increased results and absolute values of IL-8 levels showed tendency to significant increase with the stage of disease. These results suggested that IL-8 measurement may be useful in estimation of disease progression in women with breast carcinoma. PMID- 12557435 TI - [Usefulness of CEA, CA 15-3 and CA 125 tumor markers in the differential diagnostics of peritoneal effusion]. AB - To differentiate malignancy-related from non-malignant exudate is important for cancer diagnosis, determining clinical stage of the disease, establishing treatment strategy, and also for prognosis. Carcinoembryonic antigen (CEA), CA 15 3 and CA 125 levels were analysed in plasma and in peritoneal fluid of 37 cancer patients (13 with breast cancer, 18 with ovarian cancer and 6 with colorectal cancer) and 15 patients with peritoneal fluid not related to malignancy. CEA, CA 15-3 and CA 125 were measured by electrochemiluminescence assay (ECL). The results were correlated with cytological and histological findings as well as clinical data. Initial observations indicate that the analysis of tumour markers CEA, CA 15-3 and CA 125 may be useful in the differential diagnosis of peritoneal effusion. PMID- 12557437 TI - [Mediastinal tumors in children]. AB - Mediastinal tumours are not very common among various tumours in children. The aim of our study was to evaluate the clinical features, diagnostic approaches and surgical management in 125 patients treated in our Department for 30 years. There were 39 lesions in the anterior, 21 in the medial and 49 in the posterior mediastinum. Solid tumours were diagnosed in 78% of patients. The range of age was between 4 days and 18 years of life. Patients below 5 years of life accounted for 68%. The most common were neurogenic tumours--35%, less frequently tumours of lymphatic origin were seen. Complete excision was possible in 66% of patients, in 26% open biopsy examinations were performed. Enlarged thymus was under control in our out-patient clinic. Combined treatment (radio-, chemotherapy and surgical treatment) was introduced to deal with neurogenic tumours in children. Complete remission lasting 3 years was achieved in 71% of patients in this group. Comparing two periods of time (1970-85 v 1986-2000) no significant differences were found in the prevalence of any kind of mediastinal tumours. PMID- 12557438 TI - [Tissue factor (TF) and inhibitor (TFPI) concentrations in patients with urinary tract tumors and haematological malignancies]. AB - The aim of study was to evaluate TF activity and TFPI concentration in patients with haematological malignancies and urinary tract tumors. TFPI concentration and activity and TF concentration were measured in 20 patients suffering from acute myeloblastic leukaemia (AML), 21 patients with chronic myelogenous leukaemia (CML), 17 patients with chronic lymphatic leukaemia (CLL), 16 patients with multiple myeloma (MM) and 65 healthy adults. TFPI and TF concentrations were measured also in patients with renal cell carcinoma (n = 12) and bladder cancer (n = 17) and patients with benign prostatic hyperplasia (BPH) (n = 15). Patients with AML, CML, CLL, and cancer revealed elevated TFPI concentrations. Patients with AML, CML, CLL, MM showed decreased TFPI activity. However TFPI concentration correlated inversely with TFPI activity only in the AML group. No significant changes were observed in TF concentrations in all investigated groups. PMID- 12557439 TI - [Plasma selenium concentration, glutathione peroxidase and glutathione S transferase activities in patients with chronic liver diseases]. AB - The effects exerted on hepatocytes by alcohol metabolites, drugs or other toxins and also hepatotropic viruses lead to chronic liver diseases. Reactive oxygen species (ROS) have been implicated in a number of pathologies, including different types of liver diseases. Organism has developed several mechanisms to counteract or prevent reactive oxygen species effects. These include enzymes such as: glutathione peroxidase (GSH-Px) with selenium (Se) in the active site and glutathione S-transferase (GST). Measurement of GST, compared with alanine aminotransferase (AIAT), has been advocated as a superior marker of hepatocellular damage. The aim of this study was to assess selenium concentration, glutathione peroxidase and glutathione S-transferase activities in plasma of patients with various types of liver diseases. The study population consisted of 54 patients and 25 healthy volunteers. The patients were divided into two groups according to etiology of the disease. Plasma selenium concentration was reduced in patients with cirrhosis, as compared to controls, irrespective of etiology and activity of AIAT. Plasma GSH-Px activity was significantly lower in both groups of patients with normal AIAT activity, whereas it was higher in both groups with activity of AIAT higher than 40 U/l. GST activity was higher only in post-viral group in patients with high AIAT activity. Impaired intestinal absorption and distribution of selenium among plasma proteins have been suggested as possible mechanism of reduced selenium concentration. Changes in the activities of glutatthione-dependent enzymes in plasma may arise from increased formation of reactive oxygen species or from release of these enzymes from injured hepatocytes to plasma. PMID- 12557440 TI - [Comparison of ADA and WHO diagnostic criteria for diabetes diagnosis and other categories of glucose intolerance ]. AB - New diagnostic criteria for diabetes mellitus announced by ADA in 1997 and WHO in 1998 recommend lowering of the fasting plasma glucose to 7.0 mmol/l and introduce new category: impaired fasting glucose. In contrast to ADA, WHO recommends to maintain oral glucose tolerance test in clinical practice. In this study, based on oral glucose tolerance test results and WHO 1998 criteria we estimated diagnostic value of fasting glucose in diagnosing diabetes. The aim of the study was also to compare prevalence of diabetes and other categories of glucose intolerance according to ADA 1997, WHO 1985 and WHO 1998 criteria. METHODS: Oral glucose tolerance test (OGTT) was performed in 1528 patients (785 men, 743 women). According to ADA 1997, WHO 1985 and WHO 1998 criteria the patients were divided into groups depending on glucose tolerance abnormalities. RESULTS: Prevalence of diabetes according to ADA 1997 and WHO 1985 criteria was almost the same, being 10.47% and 10.34%, respectively, and according to WHO 1998 criteria it was 13.15%. In 20.4% of all the patients with diabetes according to WHO 1998 criteria, the diagnosis was made on the basis of 2-h post load glucose, in case of WHO 1985 criteria the percentage was 39.9%. 13.04% of patients with IFG according to ADA had diabetes and 34.06% had IGT according to WHO 1998 criteria. The highest overall percentage agreement (97.19%) was found between WHO 1998 and WHO 1985 classifications with the hypothesis that unclassified group according to WHO 1985 corresponds to IFG group according to WHO 1998. CONCLUSIONS: Despite lower diagnostic value of 2-hour post-load glycaemia in diagnosing diabetes according to WHO 1998 criteria compared to WHO 1985 criteria, OGTT remains irreplaceable diagnostic tool, especially in high risk populations. In patients with IFG according to ADA OGTT causes significant improvement in detecting diabetes. PMID- 12557441 TI - [Echinococcosis in North-Easter Poland--epidemiological and clinical aspects]. AB - The incidence was assessed of antibodies against E. granulosus and chosen biochemical parameters among the inhabitants of north-eastern Poland. Among 532 studied subjects, 110 were seropositive. Group I (n = 73) was pharmacologically treated. Group II was surgically treated. Control group consisted of 15 healthy persons. 20.6% seropositive inhabitants of north-east Poland indicate that echinococcosis is not rare. Mainly it was echinococcosis of the liver, only in 2.7% it was connected with other organs. Echinococcosis was diagnosed among women aged 47-70, living in town, in majority owners of a dog. The course of echinococcos in majority cases was without clinical signs. In the diagnosis USG and serological test were helpful. Laboratory findings were not characteristic of echinococcosis. If it is possible surgical treatment is recommended for removal of parasite, verification of preoperative tests. In majority of chirurgically removed cyst, antibodies against E. granulosus were found after 3.5 years on the average, what evidenced strong immunogenicity of the parasite. PMID- 12557442 TI - [The attitude of women in reproductive age toward breast self-examination]. AB - The aim of the study was to answer the question--do women at procreative age practise breast self-examination, how often do they make such examination and what are the purposes? The study was carried out in 149 female students from the Faculty of Nursing and Health Studies at the University Medical School in Lublin. The study was carried out on the basis of author's questionnaire. The results showed that 79.9% of all respondents practise breast self-examination but not all of them regularly. Well-off women more often practise breast self-examination than the poor ones. Concern about their own health was the main cause for breast self-examination. Women who have not practiced this examination said they have not been doing it because they were afraid of noticing some changes. PMID- 12557443 TI - [Modification of innate immunity in humans by active components of shark liver oil]. AB - See fish oils affect different systemic reactions innate immunity including. Innate immunity is responsible for immediate pathogen recognition and inactivation. Innate immunity decides also on the type of required immunity development. In the presented paper we have proved that supportive treatment with shark oil components normalize complement level, natural killer cells activity and reactive oxygen intermediates production by peripheral blood leukocytes of peoples suffering from active form rheumatoid arthritis. PMID- 12557444 TI - [Primary splenic angiosarcoma]. AB - Primary splenic angiosarcoma is a rare neoplasm. The symptomatology is highly variable, the diagnosis can be made usually at autopsy. The prognosis of the tumor is uniformly poor. Splenectomy prior to rupture prolongs survival. The authors describe the case of primary splenic angiosarcoma in an 54-year-old man and some diagnostic problems connected with it. The symptomatology causes diagnostic difficulties. The patient only presented weakness, enlargement of circumference of the abdomen and high hypochromic anaemia. The diagnosis was made premortem at the time of splenectomy, as a result of histological examination of tumor. Autopsy 4 weeks later revealed metastases to the liver, peritoneum, omentum which caused haemoperitoneum. Our case confirms uniformly poor prognosis of primary splenic angiosarcoma. PMID- 12557445 TI - [Pyoderma gangrenosum--positive effect of cyclosporin A therapy ]. AB - Although pyoderma gangrenosum is a disorder known since over 70 years, it still remains a diagnostic and therapeutic problem. We describe three subjects with pyoderma gangrenosum; two were females, one was male, one case was associated with colitis ulcerosa, two were without any related disorders. Histopathologic examinations supported the diagnosis in all cases. In spite of intensive topical and systemic treatment with corticosteroids, Dapsone, Clofazimine, no sufficient effects were achieved. Cyclosporin A introduced in the dose of 5 mg/kg/d resulted in dramatic response and complete remission. Serum CyA levels, biochemical parameters of liver and kidney function, blood pressure were monitored during the therapy. No adverse events due to Cyclosporin A were observed. PMID- 12557446 TI - [Radioimmunoguided surgery in colorectal cancer]. AB - Radioimmunoguided surgery (RIGS) is a technique that enables to determine the extent of a primary, as well as of a recurrent tumour and its local and distant spread. Before the surgery the patient is administered with radiolabelled monoclonal antibodies targeted against the tumour-associated antigen. The radiotracer and, in consequence, the tumour cells localisation is detected intraoperatively using a hand-held gamma detecting probe. Local assessment of tumour, regional lymph nodes or other organs (particularly liver), may allow a more complete surgical clearance of carcinoma lesions. This article presents the idea of RIGS technique (use of monoclonal antibodies, isotopes, and gamma detecting probe) and the results of worldwide clinical investigations conducted during the last years. PMID- 12557447 TI - [The role of adhesion molecules in cancer]. AB - Adhesion molecules are cell surface proteins responsible for normal structure and function of tissues and organs. The process of tumour growth often involves alterations in expression of adhesion molecules that may lead to destruction of tissue architecture and to metastasising. This study describes behaviour of some adhesion molecules (selectins, integrins, cadherins and CD44 isoforms) in different cancers. We suggest the important role of adhesion molecules in the growth of primary and metastatic tumours and we think that precise knowledge of biological activity of these molecules may lead to development of new diagnostic methods and future immunotherapy of cancer. PMID- 12557448 TI - [Anal endosonography in the diagnosis of anal neoplasm]. AB - Anal endosonography is a highly accurate imaging method in the diagnosis of rectal carcinoma. Its ability to visualise the layered structure of the anal canal enables local staging of the tumour of the anal canal. Also, the assessment of the perirectal tissues by means of endosonography, is useful in visualisation of enlarged lymph nodes. In patients after radiotherapy or chemotherapy, anal endosonography is used as the follow-up examination. PMID- 12557449 TI - [Driving license for diabetic patients]. AB - The right to be able to drive an automobile is not only one of the most cherished manifestations of adulthood, but increasingly a necessity in modern life. However, it is important to remember that in most countries, driving license for diabetic patients is generally a privilege rather than a right. PMID- 12557450 TI - [Otolaryngology in Warsaw after the November Insurrection]. AB - After the decline of the November Insurrection the Warsaw Royal University was closed. An important role of surgical ward at the Infant Jesus Hospital in the development of the Warsaw otolaryngology is pointed out. The professional and scientific activities of Aleksander Antoni Le Brun (1803-1868), who was the head of the surgical ward at the Infant Jesus Hospital are described in detail. He was a well-known Warsaw physician and an eminent surgeon. The Le Brun's achievements in the surgical treatment of otolaryngological diseases are strongly pointed out. The scientific and professional activities of such surgeons as Franciszek Groer (1807-1876), Ludwik Chwat (1831-1914) and Konstanty Miller (1823-1890) from the Orthodox Jewish Hospital and the Evangelical Hospital are described in detail. The otolaryngological operations performed in Warsaw by Bernhardt R. Langenbeck from Berlin are also depicted. PMID- 12557451 TI - The effects of stimulant therapy, EEG biofeedback, and parenting style on the primary symptoms of attention-deficit/hyperactivity disorder. AB - One hundred children, ages 6-19, who were diagnosed with attention deficit/hyperactivity disorder (ADHD), either inattentive or combined types, participated in a study examining the effects of Ritalin, EEG biofeedback, and parenting style on the primary symptoms of ADHD. All of the patients participated in a 1-year, multimodal, outpatient program that included Ritalin, parent counseling, and academic support at school (either a 504 Plan or an IEP). Fifty one of the participants also received EEG biofeedback therapy. Posttreatment assessments were conducted both with and without stimulant therapy. Significant improvement was noted on the Test of Variables of Attention (TOVA; L. M. Greenberg, 1996) and the Attention Deficit Disorders Evaluation Scale (ADDES; S. B. McCarney, 1995) when participants were tested while using Ritalin. However, only those who had received EEG biofeedback sustained these gains when tested without Ritalin. The results of a Quantitative Electroencephalographic Scanning Process (QEEG-Scan; V. J. Monastra et al., 1999) revealed significant reduction in cortical slowing only in patients who had received EEG biofeedback. Behavioral measures indicated that parenting style exerted a significant moderating effect on the expression of behavioral symptoms at home but not at school. PMID- 12557452 TI - Changes in physiological arousal to gambling cues among participants in motivationally enhanced cognitive-behavior therapy for pathological gambling: a preliminary study. AB - Despite somewhat high attrition and relapse rates, cognitive-behavioral interventions for pathological gambling seem promising. As a possible remedy to these problems, we conducted a preliminary study of gambling-specific cognitive behavior therapy (CBT) with the addition of motivational enhancement techniques (MET) for the treatment of pathological gamblers. Data on psychophysiological arousal upon exposure to imagined gambling vignettes were collected at both pre- and posttreatment. Results indicate that participants showed decreases in degree of arousal during the vignettes from pre- to posttreatment. There was also a strong dose-response relationship between reductions in gambling symptoms and reductions in arousal. These findings are discussed, as are their implications for further study of pathological gambling. PMID- 12557453 TI - EEG signature and phenomenology of alpha/theta neurofeedback training versus mock feedback. AB - Alpha/theta (a/t) neurofeedback training has in the past successfully been used as a complementary therapeutic relaxation technique in the treatment of alcoholism. In spite of positive clinical outcomes, doubts have been cast on the protocol's specificity when compared to alternative relaxation regimes. This study investigated the basic tenet underlying the a/t training rationale, that accurate a/t feedback representation facilitates the generation of these frequency components. Two groups of healthy volunteers were randomly assigned to either (a) real contingent a/t feedback training or (b) a noncontingent mock feedback control condition. The groups were compared on measures of theta/alpha (t/a) ratios within and across training sessions, as well as activational self report scales after each session. The contingent a/t feedback group displayed significant within-session t/a ratio increments not evident in the mock control group, as well as higher overall t/a ratios in some but not all of the training sessions. No differences were found between the groups in terms of subjective activational phenomenology, in that both groups reported significantly lower levels of activation after training sessions. The data demonstrate that irrespective of considerations of clinical relevance, accurate a/t neurofeedback effectively facilitates production of higher within-session t/a ratios than do noncontingent feedback relaxation. PMID- 12557454 TI - Task Force Report on methodology and empirically supported treatments: introduction. AB - Recent criticism of biofeedback has increased the importance of rating the efficacy of each biofeedback and neurofeedback therapy. A joint task force of the Association for Applied Psychophysiology and Biofeedback (AAPB) and the Society for Neuronal Regulation (SNR) has developed standards for efficacy research methodology and template for rating the level of efficacy of each application. The Task Force Report has been approved as a policy guideline by both the AAPB and SNR Boards. PMID- 12557455 TI - Template for developing guidelines for the evaluation of the clinical efficacy of psychophysiological interventions. AB - An essential function of both the Association for Applied Psychophysiology and Biofeedback (AAPB) and the Society for Neuronal Regulation (SNR) is the systematic evaluation of psychophysiological interventions that have been developed for the treatment of medical and psychiatric disorders. In order to address scientific concerns regarding the efficacy of specific clinical applications of biofeedback, these two societies formed and Efficacy Task Force. The process to be used in the assessment of treatment efficacy, specificity and clinical utility is presented in the form of a template that will serve as the foundation for a series of scientific reviews and practice guidlines to be published by both societies. PMID- 12557456 TI - The circle of the soul: the role of spirituality in health care. AB - This paper examines the critical attitude of behavioral professionals toward spiritual phenomena, and the current growing openness toward a scientific study of spirituality and its effects on health. Health care professionals work amidst sickness and suffering, and become immersed in the struggles of suffering persons for meaning and spiritual direction. Biofeedback and neurofeedback training can facilitate relaxation, mental stillness, and the emergence of spiritual experiences. A growing body of empirical studies documents largely positive effects of religious involvement on health. The effects of religion and spirituality on health are diverse, ranging from such tangible and easily understood phenomena as a reduction of health-risk behaviors in church-goers, to more elusive phenomena such as the distant effects of prayer on health and physiology. Psychophysiological methods may prove useful in identifying specific physiological mechanisms mediating such effects. Spirituality is also a dimension in much of complementary and alternative medicine (CAM), and the CAM arena may offer a window of opportunity for biofeedback practice. PMID- 12557457 TI - Fall prevention & intervention in home care. AB - According to information gathered by the Centers for Disease Control, falling is the leading cause of death in women and the fourth leading cause of death in men between the ages of 65 and 85. It is the leading cause of death for both men and women over the age of 85. The percentage of falls among community-dwelling elderly persons increases from 25 percent at age 70 to 35 percent after age 75. A comprehensive, multidisciplinary treatment plan must be established to address the manifold problem of falls in our elderly patient population. A comprehensive evaluation to include medical history, medications, functional assessment, and home safety must be performed in order to reduce the patient's risk of falls, thereby maximizing safe, functional mobility. PMID- 12557458 TI - A caregiver's eye on elders with low vision. AB - Vision loss is ranked third behind arthritis and heart disease as the most common condition causing a need for assistance with activities of daily living (ADLs) for persons 70 years and older, but is often overlooked in the home care setting when treating patients for other conditions. Visual impairments lead to patient depression, anxiety, increased passivity, decreased confidence, and feelings of social isolation. Vision rehabilitation can play a dramatic part in keeping these patients healthy and functioning at their maximal level. PMID- 12557459 TI - Respiratory therapists in home care. AB - Since the implementation of the Medicare prospective payment system (PPS) in October 2000, many sources indicate that the average numbers of patient visits in a home care episode have decreased. Patients with respiratory system diagnosis are a significant percentage of the total episodes in home care. Respiratory therapists (RTs) can provide additional staffing, meet patient needs, and control costs through greater flexibility in resource utilization. PMID- 12557460 TI - Speech pathology & Parkinson's disease in the home environment. AB - Those with Parkinson's Disease often suffer speech limitations distinct from other neurological diseases and stroke. Symptoms of dysarthria, the clinical term of impaired, uncoordinated, and weakened speech musculature, include reduced volume, breathiness, monotone pitch, accelerated rate of speech, and inability to communicate effectively over the telephone, in groups, and even in quiet one-to one settings. The author discusses the voice, speech, and related deficits encountered by individuals with Parkinson's Disease and the speech-language pathologist's role in addressing these deficits. PMID- 12557461 TI - A nutritional care primer for the elderly home care patient. AB - The need for proper nutrition exists for every home care patient. This need is heightened when trauma, illness, or disease challenges the body. Home care professionals can have a positive impact on patient outcomes by remaining alert to key nutritional issues. The author shares tips and includes a nutrition checklist created by the Nutrition Screening Initiative to help health care providers recognize the warning signs of nutritional risk. When the entire home health team works toward meeting the very basic physiological needs for adequate nourishment, patient outcomes cannot help but be enhanced. PMID- 12557462 TI - Integrating the principles of energy conservation during everyday activities. AB - Energy conservation and activity efficiency using pulse oximeter readings and adaptive techniques during activities of daily living (ADLs) can be beneficial to individuals afflicted with chronic obstructive pulmonary disease (COPD). Theory and practice are alternately explored so as to show an occupational therapy plan individualized to help the client remain safe and independent in her home. PMID- 12557463 TI - Speech therapy in a home care setting. Three case studies. AB - Home care speech-language pathologists provide individualized care with freedom and autonomy combined with the personal relationships developed between patients, their families, and the speech pathologist that are not found anywhere else. The authors present three patient cases and their outcomes. PMID- 12557464 TI - The challenges & rewards of caregiver training. AB - Home care is no longer the stepchild of the rehabilitation delivery system. It now attracts the best clinicians the field has to offer. Understanding the dynamics of the patient-caregiver relationship is critical, as is integrating the strengths and limitations of the caregiver into the rehabilitation program. The experience of the physical therapist (PT) and his or her ability to understand and respect the needs and the strengths of these individuals can result in a comfortable working relationship and a higher level of function and quality of life. PMID- 12557465 TI - Montessori-based training makes a difference for home health workers & their clients. AB - Home care visits can last several hours. Home care workers are often at a loss on how to fill time spent in homes of clients. The challenge is how to use this time in ways that are productive and engaging for both clients and home health workers. The authors trained home health aides to implement Montessori-based activities while interacting with clients who have dementia. The results were amazing. Among other positive results, the authors found a statistically significant increase in the amount of pleasure displayed by clients after health workers received training. PMID- 12557466 TI - 2002 hospice legislative regulatory summary . PMID- 12557467 TI - Combatting medication errors in home health. AB - Home care patients are vulnerable to medication errors. Until recently, however, little was done to find out how many or what types of errors were occurring. A 2001 study shows that nearly one out of three elderly home care patients has a potential medication error. The good news is that a simple intervention can be implemented by home care agencies to reduce this daunting statistic. PMID- 12557468 TI - Immunization registry progress--United States, 2002. PMID- 12557469 TI - Alcohol for hand hygiene: new comparative studies add to the evidence base. PMID- 12557471 TI - Worker health in refuse-derived fuel plants, a five-year followup. AB - Personal and area air monitoring data for endotoxins, contaminants of concern in the solid waste industry, were analysed to determine if recommended levels were exceeded and if problematic trends existed in comparison with previous measurements. Current and historic lung function and symptom information from the workforce was analysed to determine if values were within predicted norms and to observe any trends that might indicate work related ill health. Average personal exposures to endotoxins were well within recommended levels and remain unchanged from previous sampling. Lung functions in the workforce are within predicted normal values. No trends indicating reductions in lung function based on length of employment were observed and average values are in normal ranges. PMID- 12557470 TI - The effect of cycloheximide on butyrylcholinesterase activity in vivo. AB - The paper describes the catalytic activity of butyrylcholinesterase (BuChE) measured in plasma, liver, white adipose tissue, heart, and brain of rats intraperitoneally administered a single non-lethal dose of cycloheximide (2.0 mg/kg body weight; CHM). The BuChE assay was performed on rats of both sexes either administered CHM or saline (controls), and killed 2, 3, 4, 5, 10 days later. A significant decrease of BuChE catalytic activity was observed in all tested tissues except plasma. In animals of both sexes, the lowest BuChE catalytic activity was found in the liver (2-6%), while it was higher in white adipose tissue, heart, and brain. However, the respective values remained significantly different from controls (33-67%, 49-62%, and 14-71% in males, and 24-82%, 72-86%, and 33-67% in females). Since there was no effect of CHM on BuChE catalytic activity in plasma, the data suggest that CHM inhibits the synthesis of BuChE rather than its active site. PMID- 12557472 TI - Determination of benzene in urine by static headspace gas chromatography. AB - This paper describes the application of static headspace gas chromatography in determining benzene in urine. The method was analytically validated for sensitivity (DL = 42 ng/l), repeatability (RSD = 3% and 4%), and accuracy (71%), and was applied in measuring urine benzene in nonsmokers (N = 14) and smokers (N = 18). All urine samples had measurable benzene concentrations. The method proved sensitive enough to establish a significant statistical difference (P < 0.000614) in urine benzene concentrations between smokers (mean = 760; range = 181-1869 ng/l) and nonsmokers (mean = 214; range = 61-515 ng/l). PMID- 12557473 TI - The asbestos dilemma: II. The ban. AB - This article reviews the history of legal steps which led to conflicting regulatory approaches to the use of asbestos between the EU, the US and the rest of the world. Particular attention is paid to the EU ruling out the use of all types of asbestos. The author criticizes how the Scientific Committee on Toxicity, Ecotoxicity and the Environment of the European Commission altered their own opinion within as short a time as seven months, and points out the weaknesses of the scientific justification for the EU ban. With equal criticism the author addresses the weak scientific rationale on which Collegium Ramazzini founds its call for an immediate and total international ban of the mining and use of all types of asbestos. PMID- 12557474 TI - Hair testing for drugs of abuse: a two-year experience. AB - This article gives a brief account of a two-year experience with gas chromatography/mass spectrometry methods developed for the analysis of opiates (morphine, codeine, heroin and 6-acetylmorphine), cocaine, methadone, and amphetamines (amphetamine; methamphetamine; 3,4-methylenedioxyamphetamine--MDA, and 3,4-methylenedioxymethamphetamine--MDMA, Ecstasy) in hair. The methods developed were reproducible (RSD = 5.0-16.1%), accurate (85.1-100.6%) and sensitive (LD = 0.05-0.30 ng/mg). They were applied in the analysis of 221 hair samples obtained from young subjects aged 15-25 years, who were suspected of drug abuse. Seventy-seven hair samples were found positive for drugs of abuse. Forty two subjects were found to have consumed amphetamine, of whom 40 were found MDMA (Ecstasy). Heroin consumption, based on positive 6-acetylmorphine, was found in 26 subjects. Cocaine alone was present in three subjects, two were found cocaine and heroin, two cocaine and methadone, and one cocaine and MDMA. These results could indicate the trend in drug abuse among young people in Croatia. PMID- 12557475 TI - [How to write]. AB - In spite of a popular belief that every scientist knows how to write a professional paper, it is a sad fact that only a few scientists are really good writers. Hence the need for this paper. The process of writing is divided in three general steps (preparing, writing, and editing). In the first step, it is necessary to comprehend the future text as the whole. In the next step, one has to deal with its composition. In the third step, it is important to divide editing in precisely defined actions (adding and checking data, grammatical and stylistic corrections, spell-checking). The article also addresses certain differences between English and Croatian related to the meaning of words of Latin origin and provides examples of stylistic editing of scientific texts. PMID- 12557476 TI - Prevention of exposure to mycotoxins from food and feed. AB - Mycotoxins are metabolites of moulds that may be found in food and feed of plant and animal origin. This paper gives a short review of the agronomical methods and food and feed storage recommendations for the prevention of mould contamination. It describes the practical methods of feed decontamination and the use of feed additives where mycotoxin contamination prevention has failed. However, these methods should be avoided as much as possible because they may increase the cost of production, reduce the nutritional value of feed, and leave residues of mycotoxins or their toxic metabolites. Since there is no universal and reliable method of feed decontamination for all mycotoxins, the paper stresses the importance of preventive measures. PMID- 12557477 TI - [Activity of the corn Spm transposon system in transgenic plants Orychophragmus violaceus (L.) O.E. Schulz obtained by both direct transfer of DNA to protoplasts and agrobacterial transformation of root explants]. AB - Transposon mediated insertional mutagenesis is one of the approaches for the unique gene cloning. A wild species of Cruciferae family Orychophragmus violaceus (L.) O.E. Schulz, which is of interest for practical breeding as a donor of improved plant oil, was an object of the investigation. Plasmid construction used in the experiments included selective NPT II gene, reported GUS gene serving as an excision marker, structural BAR gene located within the dSpm element and Spm transposase. The GUS gene of this plasmid had not his own promoter and became functional only after Spm-transposition. Transformed Orychophragmus violaceus (L.) O.E. Schulz. plants were obtained by direct mesophyll protoplast transformation as well as Agrobacterium tumefaciens-mediated root explant transformation. Gene transfer and the transposition event were confirmed by the GUS activity and the PCR analysis. Relative transformation efficiency using protoplasts was 5.8%. PMID- 12557478 TI - [DNA-marking of quantitative traits in corn]. AB - Methods of ISSR- and RAPD-analyses were used for marking quantitative trait loci (QTLs) determining the development of some morphological and biological traits in maize. Specificity of marker locus alleles was established for certain levels of polygenic trait phenotype manifestation. Criteria of marker locus informativity are discussed. A possibility of marker-assisted selection for valuable genotypes with desired for breeding trait values was demonstrated. PMID- 12557479 TI - [Genetic-statistical characterization of the novel different-height donors of the stem shortness in winter rye]. AB - Three donors for the dominant stem shortness in winter rye were developed. The genetic determination of the difference in a plant height for each donor was revealed. There were additive genes for the stem shortness in the donors Gnom 1 and Gnom 2 and dominant-epistatic ones in the donor Gnom 3. The investigation of genetic correlation between the number of quantitative traits and the plant height was carried out. On the basis of the data obtained, the strategy of using each donor for stem shortness in rye breeding was determined. PMID- 12557480 TI - [Self-fertility problems in birdsfoot trefoil (Lotus corniculatus L.)]. AB - The problems of Lotus corniculatus self-fertility are analyzed. The self compatibility of hereditivity character, the efficacy of different methods of self-pollination (autogamy, geitonogamy within one plant and among plants of one clone), and usage of the mixture of self-compatible and self-incompatible pollen were studied. PMID- 12557481 TI - [The investigation of genomes of some species of the genus Gentiana in nature and in vitro cell culture]. AB - The comparative study of the genomes of intact plants-representatives of some species of the genus Gentiana L. as well as cultured cells of G. lutea and G. punctata was performed using restriction analysis. Species specificity of restriction fragment patterns for studied representatives of this genus was revealed. The differences between electrophoretic patterns of digested DNA purified from rhizome and leaves of G. lutea and G. punctata were found. The changes in genomes of G. lutea and G. punctata cells cultured in vitro compared with the genomes of intact plants were detected. The data obtained evidence that some of them may be of nonrandom character. PMID- 12557482 TI - [A new gene for the grown together florets in corn]. AB - A new mutation for maize tassel structure was founded. This mutation is controlled by a recessive gene. It was named "tassel knit"(tk). The new gene can be used for corn hybridization. PMID- 12557483 TI - [Cytogenetic effects in children exposed to low doses of ionizing radiation]. AB - The obtained data indicate that frequencies of different types of cytogenetic anomalies in investigated children groups living in radionuclide contaminated territories and children irradiated in utero have complicated patterns. The frequency of chromosomal anomalies in the investigated groups of children exceeds the average population level. At the same time, no statistically significant differences in frequencies of various types aberrations between groups of children were revealed. PMID- 12557484 TI - [To the problem of differentiation of north Mongoloids, Caucasoids, and Amerinds on the territory of Eurasia (genetic data)]. AB - The matrix of genetic distances of 11 human populations of Europe, Asia, and America was calculated using data on frequencies of 28 alleles of 12 loci of proteins, enzymes, and blood groups. This matrix and constructed on their basis dendrogram permit one to suggest that the place of division of paleolitic population on ancestors of North Mongoloids, Caucasoids, and Amerinds was in South Siberia and possibly in neighbouring regions of Central Asia. Published data on polymorphism of mtDNA in the human populations of Eurasia and America were analysed. These data support the author's hypothesis. PMID- 12557485 TI - [Sex ratio in Down syndrome]. AB - Data from 55 publications providing the sex ratio (SR), i.e. ratio between male and female cases of Down syndrome (DS), are presented. In general, SR was skewed toward an excess of males in the majority of studied populations, either in populations with a high level of cases ascertainment (epidemiological studies) or in selected groups. No significant correlation involving the age of either patients or mothers was found. Some other factors which might influence the sex ratio in DS at birth are mentioned. Meta-analysis of data from epidemiological studies suggests the phenomenon is not restricted to free trisomy 21 alone but appears in translocation cases, both in mutant and inherited translocation carriers (SR = 1.31 and 1.36, respectively). In contrast to nonmosaic 47, +21 cases, where SR is close to 1.3, an excess of females was observed in mosaics 46/47, +21 (SR = 0.83). No male predominance was found among patients with DS not tested cytogenetically (SR = 0.98), which may be explained by female predominance in false-positive cases. In populations with a fraction of clinically diagnosed cases of 30% and over, SR has intermediate value of 1.1. The ratio showed a tendency to increase since 1940's, reaching a mean value of 1.35 in 1980's varying from 1.3 to 1.62 in different populations), which might be a consequence of the growing use of karyotyping to confirm diagnosis and of a real increase in proportion of males. In the 1990's, the ratio fell to 1.22 varying from 1.03 to 1.27. As SR is assumed to reflect a proportion of paternal contribution, the discrepancy between the proportions of paternal errors in cytogenetic studies on parental origin of the extra chromosome (24% in the 1980's) and in molecular studies (5-10% in the 1990's) discussed in the literature might be explained by temporal changes alone. Genetic mechanisms of male predominance in trisomy 21 are reviewed, among them models for joint segregation of chromosome 21 and Y chromosome in spermatogenesis, and the chromosome 21 nondisjunction during 2nd meiotic division of oogenesis caused by Y chromosome-bearing spermatozoa. PMID- 12557486 TI - [Problems of formation of genetic coadaptation during creation of synthetic genotypes]. AB - In the review the problems of restoration of genetic coadaptation in some agricultural crops under creation of synthetic genotypes using sexual hybridization and transgenosis are considered. On the basis of the study of multiple molecular forms of the enzymes, the mechanisms of formation of adaptive complexes of genes (ACG) are traced. Cytogenetic mechanisms interfering stabilization of synthetic genotypes are considered. PMID- 12557487 TI - [Viability of lyophilized cells of Myxococcus xanthus UKM 10041 and Polyangium cellulosum UKM 10043 in presence of different antioxidants]. AB - It has been established that addition of antioxidants: cystamine, ionol, alpha tocopherol to protective media (saccharose-gelatin agar) does not affect considerably viability of lyophillized cells of cultures Myxococcus xanthus UCM 10041 and Polyangium cellulosum UCM 10043. Experimentally obtained and predicted data on the survival of cells of M. xanthus UCM 10041 and P. cellulosum UCM 10043 are the values of the same order which evidences for the possibility of the use of quick test for prediction of myxobacteria cells survivability under long-term storage. PMID- 12557488 TI - [Lectin activity of antitumor substances synthesized by Bacillus subtilis B 7025]. AB - Nutrient medium, which is optimal for synthesis of biologically active substances and allows obtaining stable outflow of Bacillus subtilis B-7025 to the culture medium, has been selected under the conditions of the bacillus periodic cultivation. It has been established that these substances are extracellular lectins with high activity and carbohydrate specificity to fructose-1.6-di phosphate, N-acetylneuraminic acid and glucose derivatives: D-glucuronic acid and D-glucosamine. PMID- 12557490 TI - [The distribution of introduced nitrogen-fixing bacteria in the rice root zone]. AB - The most oxidized zone 0-1.5 mm of root, which determines the high number of aerobic diazotrophic microorganisms, the reduced zone 1.5-4.5 mm, where their quantity was decreased and maximum of anaerobic nitrogen-fixing bacteria development observed as well as the reducing zone 4.5-7.5 mm, in which aerobic, obligate and facultative anaerobic bacteria coexist, have been discovered in the rice root exudations and their decomposition products. PMID- 12557489 TI - Selection of pectinesterase overproducing fungal strains for the low degree methoxylated pectin preparation. AB - The selection of the novel pectinesterase (PE) producing strains is an important step in their biotechnological application. The present work displays the purposeful multistage selection of strains with the increased ability to synthesize extracellular PE among 700 filamentous fungal collection cultures belonging to 45 genera and 93 species. A scheme of targeted multistage screening of PE producers has been developed. At the first stage of the screening 23 potentially active strains belonging to the genera Penicillium, Fusarium, Aspergillus, Rhizopus, Gliocladium, etc., were selected. Strains of the Trichoderma, Cochliobolus, Cladosporium and Thielavia genera not yet described as PE-active ones were selected. The P. funiculosum species (0.28-0.56 units/ml) possessed higher ability to synthesize PE. High level of the PE activity was revealed in P. rubrum (0.37 units/ml). No correlation between the level of fungal biomass accumulation on pectin-containing medium and the level of their PE activity was observed. A principal chance of the fungal PE substitution for some chemical reagents at the stage of demethoxylation of the pectin-containing raw materials should be noted. The technology of the fungal PE application for production of the low-degree methoxylated pectin is under development. Such pectin is necessary for the low-calorie jellfying products in the food industry and in medicine as a preventive agent removing the toxic substances from the human body. PMID- 12557491 TI - [Effect of decapitation on the systemic acquired resistance of hypersensitive tobacco mosaic virus]. AB - The decapitation depressed the development of the system acquired resistance to reinfection with tobacco mosaic virus in the leaves of hypersensitive tobacco plants Nicotiana tabacum L. (v. Xanthi nc) and in protoplasts, isolated from these leaves. PMID- 12557492 TI - [Effect of some electron acceptors on metabolism of methanogenic biotic community]. AB - The dynamics of redox potential, pH, CH4, H2, CO2, N2 formation under the community development under anaerobic conditions in presence of acceptors of electrons (potassium ferricyanide, molecular oxygen, sodium nitrate and sodium sulphate) has been investigated. It has been established that the optimal value at the redox potential under the development of methanogenic community was created at the expense of microorganisms vital activity. The decrease of methanogenesis under the electron acceptors introduction is connected with the increase of the culture medium redox potential. PMID- 12557493 TI - [Influence of manganese on iron accumulation and flavinogenesis in yeast Debaryomyces hansenii]. AB - In response to MnCl2 introduction the yeast Debaryomyces hansenii shows the increase of intracellular iron content and riboflavin biosynthesis level. Under iron deficiency the yeast cells sensitivity to manganese ions increases in spite of redundant synthesis of riboflavin. High concentration of iron in the culture results in the lower toxicity of Mn2+. Iron accumulation favours the cell resistance to manganese. Manganese causes the redundant synthesis of riboflavin as well as iron accumulation in streptonigrin-resistant mutants with a lowered iron content in cells. The Mn(2+)-resistant strains are phenotypically similar to the previously described Rib80(-)-mutants of D. hansenii with high iron content and riboflavin biosynthesis level. This suggests that riboflavin redundant synthesis and iron accumulation in the yeast cells are regulated by a common Mn(2+)-depending factor. A new method for isolation of the Rib80(-)-mutants is described. The method is based on the positive selection of manganese-resistant clones. PMID- 12557494 TI - [Phagocytic activity of mouse peritoneal macrophages under administration of bacterial vaccines and prospidin]. AB - Phagocytic activity of peritoneal macrophages in cases of separate and combined injections of killed bacterial vaccines and prospidin has been studied in the work. As a result of researches conducted, it has been found out that the injection of vaccines together with prospidin makes it possible to reduce an inhibiting effect of prospidin on phagocytic activity of monocyte-macrophagal cells; first of all it concerns their digestive activity. Thus, the immunodepressing effect of antiblastome drugs may be reduced with the help of injection of killed bacterial vaccines which stimulate nonspecific factors of organism protection, including antineoplastic ones. PMID- 12557495 TI - [Species composition and antibiotic sensitivity of staphylococci isolated from patients with furunculosis]. AB - Biological properties and sensitivity to modern antibacterial drugs 54 strains of staphylococci isolated under furunculosis have been studied with the aim to optimize this disease treatment. Staphylococcus aureus was revealed with frequency 35.2 +/- 6.50%, S. epidermidis (31.5 +/- 6.32%), S. warneri (16.7 +/- 5.08%), S. hominis (7.4 +/- 3.56%), S. haemolyticus and S. saprophyticus (3.7 +/- 2.57%) as well as S. lentus (1.9 +/- 1.86%) were identified among coagulase negative staphylococci. Methycillin-resistant staphylococci were isolated from 25.92% of patients with furunculosis. High level of resistance of furunculosis agents to erythromycin, clarythromycine, cephtriaxon, pephloxacyne, tetracycline, doxycycline, levomycetine and phosphomycine has been registered. Basing on the study of isolates antibioticograms, cephasoline is recommended as the selected drug under furunculosis, and tobramycin as the reserve antibiotic. Autovaccine was also made for the patients treatment. PMID- 12557496 TI - [Ivan Hryhorovych Savchenko--famous pathologic anatomist, microbiologist and immunologist (on the 140th anniversary of his birth and 70th anniversary of his death)]. PMID- 12557497 TI - [Osteogenesis of HA/TCP biphasic ceramics implanted into muscle: a long-term study]. AB - New bone formation in long-term intramuscle implant of Ca-P biomaterial was investigated in this experiment. After implanting into dog dorsal muscle for 15 months, a thin fibrous membrane that wrapped HA/TCP implant was still observed obviously. Three types of tissues, i.e. mesenchymal tissue, bone and bone marrow, regularly distributed in different pores of implant. Nearly all the pores of implants were occupied by bone. Bone in the pores located in the central region of implant was matured lamellar bone characterized by obvious lacuna and rich bone marrow. However, bone in the peripheral pores was immature woven bone without bone marrow formation. Furthermore, mesenchymal tissues only exist in the peripheral pores and usually were connected with immature woven bone. It was demonstrated that porous HA/TCP has bone inductivity and it could induce new bone formation at non-osseous site. Well-regulated distribution of mesenchymal tissue, bone and bone marrow in the pores suggest bone morphogenesis in the implant must obey a specific space-time program. PMID- 12557498 TI - [A study of bone-like apatite formation on porous calcium phosphate ceramics in dynamic SBF]. AB - This study aimed at investigating the influence of the flow rate of simulated body fluid (SBF) (2 ml/100 ml.min) of body fluid in skeletal muscle upon the formation of bone-like apatite on porous calcium phosphate ceramics. The in vitro immersion experiment in SBF flowing at normal physiological rate is referred to as dynamic SBF. The results showed that bone-like apatite could only formed in the pores of porous calcium phosphate when SBF flow at physiological rate (2 ml/100 ml.min) of body fluid in skeletal muscle. At the same time, bone-like apatite could form both in the pores and on the surface of the samples if the flowing physiological solution is 1.5 SBF. When the flowing speed of SBF is higher than normal physiological speed (10 ml/100 ml.min), no bone-like apatite could be detected both on the surface and in the pores of the materials. This result is in concordance with animal experiments. The dynamic SBF simulates the biological environment of bone-like apatite formation in body better than static SBF (SBF does not flow). This method is very useful for the research of the mechanism of bonelike apatite formation, which is the key step of bone growth on biomaterials, and can be used as an effective approach to investigate mechanism of the osteoinduction of calcium phosphate in nonosseous tissues in vivo. PMID- 12557499 TI - [MC3T3-E1 osteoblasts adhesion to micropatterned surfaces]. AB - Cell adhesion to material surface plays an important role in regulating cell function such as proliferation and differentiation. Surface patterning provides a useful method to control cell spatial distribution and adhesion to substance. Here microcontact printing and microfluidic channels were introduced to pattern MC3T3 E1 osteoblasts on silicon substance. Dichlordimethylsilane (DMS) was used in microcontact printing to generate the alternating domains of DMS and non-DMS, and cells preferentially adhered to the non-DMS and hydrophilic region. On the patterned surfaces generated from collagen and albumin solutions with microfluidic channels, cells preferentially localized in the collagen-coated region. The results also showed that micropatterning could be a useful method to study the effect of surface chemistry on cell adhesion and other functions. PMID- 12557500 TI - [Preparation of calcium phosphate coatings on surfaces of commercially pure titanium induced by simulated body fluid]. AB - All samples were divided into three groups and subjected to three different surface treatments, i.e. untreated group, group etched with mixed acid of 1:1 HCl and H2SO4 followed by immersion in 6N NaOH solution at 60 degrees C for 24 h and group etched with mixed acid of 1:1 HCl and H2SO4 followed by immersion in 6N NaOH solution at 60 degrees C for 24 h then heated at 600 degrees C for 1 h. After soaked in simulated body fluid for two weeks, a thin calcium phosphate coatings was precipitated on the surfaces of the two treated samples. Results of SEM and EDX showed that calcium phosphate coatings on the surfaces of the titanium samples etched and heated were more even than the titanium samples etched without heating. The analysis of XRD demonstrated the main component of calcium phosphate coating was hydroxyapatite. PMID- 12557502 TI - [Study on preparation and property of PLA/liquid crystal composite membranes]. AB - In this paper we synthesised two kinds of hydrophilic liquid crystal, i.e. cholesteryl triethenyl glycol carbonate (TCC), cholesteryl tetraethenyl glycol carbonate (TeCC), which were blended with PLA with certain contents to form composite membranes. Contact angle test showed that the contact angle decreases with the increase of liquid crystal. The blood-compatibility of the composite membranes was assessed by blood clotting time and hemolysis ratio which showed that these PLA/liquid crystal composite membranes with the content of 30%-40% liquid crystal appeared to be beneficial in improving the blood compatibility and anticoagulation. PMID- 12557501 TI - [Preparation of thin hydroxyapatite layers on cp titanium through anodic oxidation followed with hydrothermal treatment]. AB - To study the method of anodic oxidation followed by hydrothermal treatment for cp titanium and to know bone response to thin hydroxyapatite layers in vivo, commercially pure titanium plates were anodized at 200 V-400 V with direct electric current density no more than 50 mA/cm2 for 15 minutes in the electrolytic trough. beta-glycerophosphate sodium(0.03-0.04 M) and calcium acetate(0.2-0.3 M) were used as electrolytes. Then, titanium plates were hydrothermal treated in the autoclave for 2 hours at 280 degrees C-300 degrees C. Polishing and grit-blasting surface was used as control to learn bone response to thin layers. Twelve rabbits were evenly divided into 3 groups, each group was implanted with 12 implants into the rabbits femoral bone. After 4, 8 and 16 weeks, implants were taken out and collected respectively and were made grinding slices. The bone-implant interface was observed with light microscope. And the bone-implant interface of polishing and layered implants after 8 weeks implantation was observed with scanning electron microscope. The element contents at the interface of polishing and layered implants before and after 8 weeks implantation were detected with EDAX. Results showed that there was hydroxyapatite(HA) precipitated on the titanium surfaceamellae bone in 8 weeks for thin HA coatings, and no HA debris were found at the interfacial zone. In addition, Ca and P content on the hydrothermal treated implant surface increased much more after implantation than that of polished implants. It was concluded that anodic oxidation followed by hydrothermal treatment could precipitate thin hydroxyapatie layer on the surface of cp titanium, which could improve early formation of woven bone and accelerate woven bone transferring to lamellae bone at the implanted site. PMID- 12557503 TI - [A study on the mechanism of bFGF promoting endothelial cells to adhere to polyurethane material]. AB - bFGF can promote endothelial cell to adhere to artificial polyurethane material. However the mechanism is still unclear. Here the effect of bFGF on endothelial cell expression of integrin beta 1 subunits was determined with Western Blot and image analysis method. The result reveal that the mean gray value of immunostain is 166.1 +/- 9.86 in the experimental group and 175.32 +/- 5.12 in the control group, suggest that bFGF may upregulate the endothelial cell expression of integrin beta 1 subunits. It is concluded that the bFGF may induce the overexpression of endothelial cell integrin beta 1 subunits to increase the cell adhesion onto polyurethane material. It is suggested that bFGF can be taken as an important assistant factor in accomplishment of endothelialization on artificial heart and blood vessel. PMID- 12557504 TI - [Comparison of biomechanical properties between human nasal periosteum and fascia]. AB - There has been a lot of controversies on which layer the silastic implants should be inserted in the augmentation rhinoplasty, i.e. subperiosteal or deep subfascial. This study is to investigate the biomechanical properties of human nasal periosteum and deep fascia, including tensile strength, stress-strain and stress relaxation characters under uniaxial tension system. The periosteum is stronger in tensile strength than that of the fascia, but it is less elastic. Under a sudden increase of load, the periosteum relaxes far less than the fasia. Therefore, in view of biomechanics, the periosteum is thicker, tougher, stiffer and less relaxation than facia, thus has a better fixation effect. PMID- 12557505 TI - [Influence of flow shear stress on apoptosis of arterial endothelium in vivo]. AB - To explore the changes of wall shear stress(WSS) effect on arterial endothelial cell(EC) apoptosis after reducing arterial blood flow. The reducing flow model was established in 60 rabbits. Endothelial stretched preparations were made at 8 different time intervals from 0 to 30 days. The apoptosis rate of arterial endothelial cells (AEC) was measured with TdT-mediated dUTP-biotin nick end labeling(TUNEL) method. The results showed that the apoptosis rate of AEC was significantly higher from 1 day to 7 days after decreasing WSS than that of control, which peaked on day 3. While with progressively increasing in WSS, the apoptosis rate restored to the level of control from 14 days to 30 days. These suggest that the apoptosis state of AEC might be markedly influenced by the changes of WSS. The persist decreasing of WSS may be the important factor which induces the cell apoptosis. PMID- 12557506 TI - [A biomechanical research of neuromuscular electrical stimulation to simulate muscular force training]. AB - Based on the theoretical and experimental researches of neuromuscular electrical stimulation to simulate muscular force training, the results showed: The effect of neuromuscular electrical stimulation (NMES) is much better than that of muscular electrical stimulation (MES); The increase of muscle force is closely related to the mode and electro properties of electrical stimulation; NMES can fulfill the muscle force training with lower energy consuming, so it is practical helpful method before, during and post training muscle force training; The method of NMES simulating muscle force training can meet the synchronous and harmonious ability of muscle group. PMID- 12557507 TI - [Experiment study on stress relaxation and creep of spinal lumbar vertebrae (T12 L5) by simulating operations of excising lumbar disc intervertebrales by the front route and by the back route]. AB - We have studied the characteristics of stress relaxation and creep on lumbar vertebrae(T12-T5) of 18 fresh corpses. We measured the stress relaxation and creep of the normal group(intact spine), the control group 1(simulating the operations by the front route) and the control group 2(simulating the operations by the back route). Then we obtained the stress, strain-time curves and data under the conditions of constant stress and strain. By using regression analysis we obtained the reduced stress relaxation and creep functions. Finally, We analyzed and discussed the effects of the operations of excising the disc intervertebrales by the front route and the back route. PMID- 12557508 TI - [A perturbation solution of pulsatile Casson flow in the parallel-plate flow chamber]. AB - The parallel-plate flow chamber (PPFC) with rectangular shape, of which the height is far smaller than its own length and width, is one of the main apparatus for in vitro studies of the mechanical behavior of the cells. Considering that the Reynolds numbers of flows in the usually used PPFCs are small, a perturbation solution of laminar pulsatile Casson fluid in the PPFC is presented using Reynolds number as perturbation parameter. Furthermore, the velocities and shear stresses in the PPFC are given. The numerical results demonstrate that under the same pressure gradient the shear stresses are almost identical between Casson fluid and Newtonian fluid, whereas under the same flux the shear stresses are obviously different between Casson fluid and Newtonian fluid. The results in this paper provide a theoretical way to determine the shear stresses in the parallel plate flow chamber under pulsatile Casson fluid. PMID- 12557509 TI - [Effects of biventricular support with Luo-Ye pump on hemodynamics during ischemic biventricular dysfunction]. AB - To compare the effects on hemodynamics of univentricular support with that of biventricular support on experimental ischemic biventricular dysfunction so as to provide experimental basis for clinical usage of the Luo-Ye pump. Eight canines were placed with a left ventricular assist device (LVAD; left atrial-aorta bypass) and a right ventricular assist device (RVAD; right atrial-pulmonary artery bypass). Left anterior descending coronary artery(LAD) was ligated, three minutes later, the proximal of right coronary artery (RCA) was ligated to establish animal madel of acute ischemic biventricular dysfunction. First start the LVAD, and then RVAD was started five minutes later. The hemodynamic data were recorded including central venous pressure(CVP), cardiac output (CO), mean artery pressure(MAP), and pulmonary artery pressure(PAP) and pulmonary capillary wedge pressure (PCWP). During biventricular assist devices (BVAD) the hemodynamics were improved remarkably, MAP increased from 37.4 +/- 8.8 mmHg to 84.2 +/- 9.7 mmHg (P < 0.01) (the normal level), CO increased from 0.82 +/- 0.1 L/min to 1.33 +/- 0.12 L/min (P < 0.01), CVP decreased from 14.6 +/- 2.3 cmH2O to 4.2 +/- 1.5 cmH2O (P < 0.01), PCWP decreased significantly from 14 +/- 3.9 mmHg to 1.6 +/- 0.9 mmHg. These data suggest that LVAD during biventricular dysfunction could not improve the hemodynamics to normal level. Howere BVAD could increase CO and MAP to normal level and decrease heart work and myocardial oxygen consumption, which could help to improve myocardial metabolism and myocardial function. Therefore, BVAD is the first choice in treating severe biventricular dysfunction which was not respond to drug therapy and intra-aortic balloon pump (IABP). PMID- 12557510 TI - [The circulatory and respiratory dynamic mechanisms of lung Qi-deficiency]. AB - As the transportation of O2 and CO2 inside human body is an integrated aspect of Qi-blood transportation, a hemodynamic model is established based on the analysis of the respiratory and circulation system. The results of such model show that the abnormality of the circulatory and respiratory parameters always lead to a reduction of Po2 (O2 pressure) in tissue fluid, and then a Lung Qi-Deficiency Syndrome (QDS). This model can be used to quantitatively explain, to some extent, the physiological phenomena of Lung QDS, and could combine all the discoveries of in clinical researches, on Lung QDS. The measurement of Po2 in tissue fluid as an index to analyze Lung QDS is in accordance with TCM and TMM (the theory of modern medicine). This model may provide a new approach in clinical research of Lung QDS. PMID- 12557511 TI - [Hemodynamics monitoring of liver transplantation with color Doppler ultrasound]. AB - By hemodynamic monitoring of liver transplantation with Color Doppler ultrasound, we observed the hemodynamic changes pre- and post-operatively and the patency of the anastomotic vessels, to detect or exclude the presence of complications, then assess the prognosis. We measured the hepatic blood flow after orthotopic liver transplantation in recent 6 cases, and observed the patency of anastomostic vessels and the hemodynamic improvement of the portal vein and hepatic arteries. The abnormal liquid and solid echoes were not found around the transplantated liver in all 6 cases. The echoes of hepatic parenchyma and tububes were normal. The direction and speed of blood flow were also normal. Five of anastomoses of portal vein in 6 cases were patent. The rest one was a little stenosed, and the speed of local blood flow was a little faster. There were 11 patent ones and 1 mildly stenosed in the 12 anastomoses of inferior venae cavae. The speed index(SI) and resistance index(RI) of hepatic arterial blood flow distal to the anastomoses were all normal. Liver transplantation is the redical therapying of portal hypertension, and the color Doppler ultrasound is the best non-invasived method for monitoring the hemodynamic changes of transplantated liver. PMID- 12557512 TI - [The study of erythrocyte fragility and morphological changes caused by roller pump in vitro]. AB - To observe the erythrocyte fragility and morphological changes of erythrocytes caused by roller pump. Ten tests were divided into two groups, Polystan pediatric pump group A (n = 5) and COBE pump group B (n = 5). Ten whole blood samples (each 400 ml) were circulated in the roller pump for 16 h. Erythrocyte fragility and free hemoglobin were measured before pumping and at every 2 hours during pumping. The possible morphological changes of erythrocytes caused by roller pump were observed by scanning electron microscope. The electron microscopic observation was made before pumping and at every 4 hours throughout pumping. Results showed that the erythrocyte fragility of two groups was not increased during a long period of pumping. The number of acanthocytes of two groups was 1.77/1.81% in the samples before pumping and 6.12/7.13, 9.18/8.73, 13.21/12.89, 16.53/17.21% at 4 h, 8 h, 12 h, and 16 h respectively. The free hemoglobin level of two groups was increased linearly during a long duration of pumping and the index of hemolysis of two groups was 0.296 mg/L/h and 0.3993 mg/L/h respectively. The result shows: 1. the erythrocyte fragility was not increased during a long period of pumping; 2. the erythrocyte membrane was injured or broken by roller pump directly; 3. the morphological changes of erythrocytes would be the basis of post operative hemolysis. PMID- 12557513 TI - [The investigation of bioadhesive property of carbomer 934 by gamma camera in vivo]. AB - To study bioadhesive property of carbomer 934 in dog alimentary tract, carbomer 934 and ethylcellulose were radiolabelled with technetium-99 m. The gastrointestinal emptying rate of these materials was measured by the technique of gamma scintiscan. The results showed that the empty rate of adhesive material (carbomer 934) was remarkably slower in dog alimentary tract compared to nonadhesive material (ethylcellulose). It is concluded that, in dog, the interaction between gastrointestinal mucus layer and adhesive material or nonadhesive material was significantly different. Carbomer 934 had stronger bioadhesive property in vivo than that of ethylcellulose. PMID- 12557514 TI - [Data mining in diagnostic knowledge acquisition from patients with brain glioma]. AB - In order to correctly predict the malignant degree of brain glioma, three data mining algorithms: multi-layer perceptron network(MLP), decision tree, and rule induction are adopted to acquire diagnostic knowledge from patients with brain glioma cases. Totally 280 cases are collected, and some of them contain missing values. Preprocessing is taken to make them applicable to all three algorithms. Performance comparisons are carried out with a 10-fold cross validation test. Although the result of MLP is hard to be understood and cannot be applied directly, its reliability and accuracy are the highest when only a few hidden nodes are involved. Unlike MLP, both decision tree and rule induction use attribute-value pairs to represent diagnostic knowledge derived from treated cases. These could improve both the understandability and applicability of their results. When compared with rule induction, the inherent restriction in structure makes decision tree more efficient in decision-making but meanwhile hurts its simplicity, accuracy, and reliability. For testing samples, results of all these algorithms can achieve accuracy rate over 80%, which satisfies the basic requirement of neuroradiologists. If diagnostic accuracy rate is the main factor to be considered, MLP with only a few hidden nodes is the best. If the result is expected to be further checked or evaluated, rule induction will be the best algorithm. This work proves that data mining techniques can be used to obtain valid diagnostic knowledge from brain glioma cases and make computer aided diagnosis system in this field feasible. PMID- 12557515 TI - [Changes of subcellular calcium in hypo-themal-preserved cat kidney cortex cells detected by X-ray microanalysis of microsections]. AB - It was pointed out by many researches that keeping the concentration of Ca2+ in cells could increase the survival rate of hypothermic preserved kidneys and the survival rate of transplants. In this study, changes of the concentration of calcium were detected within catoplasm, mitochondria, endoplasmic reticulum and nucleus in the isolated hypothermic storage cat kidney, Ca2+ be marked with calcium cytochemical probe (K2H2Sb2O7) and detected by X-ray microanalysis of microsections. After 24, 48 and 72 hours preservation, the p/b (peak/back) of calcium within cytoplasm and mitochondria increased significantly. There were no obvious changes within endoplasmic reticulum and nucleus. It demonstrates that the Ca2+ were released from calcium pool (except the endoplasmic reticulum, nucleus, mitochondria etc.) to cytoplasm during preservation; and mitochondria can uptake calcium from cytoplasm to some extent, while the calcium concentration of cytoplasm is higher than normal. PMID- 12557516 TI - [Cloning and expression of heavy chain variable region genes against telomerase protein hTERT]. AB - Single domain antibodies against telomerase protein hTERT were prepared by technique of displayed on the surface of recombinant bacterio phages. Total RNA of spleen lymphocytes were extracted from mice immunized recombinant hTERT and transcripted to cDNA. First-strand cDNA was used as a template, heavy chain variable region genes against hTERT were amplified with VHfor and VHback primers by PCR technique. Amplification reaction yielded a fragment about 350 base pairs in length. Amplified cDNA were cloned into the vehide of bacteriophage PCANTAB 5E, the phagemid containing VH gene transformed into competent E. coli TG1, in the presence of helper phage M13K07, VH-g3 fusion proteins were display on the surface of recombinant phages. The phage carrying VH genes that encode binding activities could be detected directly with DOT BLOT. Single domain antibodies were generated successfully and had binding activities with hTERT. Results suggest that phage display technique be a new way of making antibodies. VH genes were cloned successfully, which could provide possibility for futher preparing single-chain antibodies(ScFv) anti-hTERT. PMID- 12557517 TI - [The experimental research of the killing effect of energy-controllable steep pulse on tumor tissue in vitro]. AB - Experimental research of injury on tumor tissue in vitro is conducted with homemade energy-controllable steep pulse device. With the comparison of histological assay results between treatment group and non-treatment group, basic phenomenon of electrochemical reaction and pathology reaction of tumor tissue during the experiment is observed. The results showed the irreversible breakdown penetrating effect of energy-controllable steep pulse on tumor cells and the feasibility of this therapy are also demonstrated. These results provide a consolidate theoretic and applicable basis for further study on mechanism and animal experiment in vivo. PMID- 12557518 TI - [Adaptive restoration of single ion channel signal under filtering and colored background noise]. AB - In order to overcome the effects of the anti-aliasing filter and the colored background noise, an adaptive algorithm is proposed to estimate the parameters of ion channel kinetics and the background noise, and whereafter the ion channel signal could be restored from the strong noisy patch-clamp recordings. The algorithm cross-couples the recursive expectation-maximization algorithm, which estimates optimally the parameters of hidden Markov model, and the recursive extended least square algorithm, which estimates optimally the characteristics of the background noise. Simulation suggests that this cross-coupling algorithm convergences consistently, and is very robust to the inexact conformation number. PMID- 12557519 TI - [Chemical QSAR recognition by using fuzzy min-max neural-network]. AB - By using the fuzzy min-max neural network, the quantitative structure-activity relationship (QSAR) of mutagenicity is studied. With the established QSAR model, the mutagenicity is predicted and the results showed that QASR is superior to linear-regression model. Further discussion on the models and the results is presented in this paper. PMID- 12557520 TI - [ECG detection method based on adaptive wavelet neural network]. AB - Adaptive wavelet neural network intelligent ECG detection is a method in which the wavelet function substitutes for the effective function of connotative layer in the neural network. Time-shift and scale parameter may be adjusted by the neural network. Therefore wavelet neural network may improve the detection rate and reliability of ECG signal. PMID- 12557521 TI - [Hidden Markov model used in protein sequence analysis]. AB - Hidden Markov model (HMM) used in the research of protein is a new field of bioinformatics. Nowadays large amount of data about protein sequences and structures have been obtained. Traditional methods of protein analysis are no longer used. Biologists have updated their research methods with computer technology and statistics, which can deal with large amount of data. HMM can be used to distinguish protein sequence with the same characteristics. A family of protein from SCOP database was selected, through which a HMM model representing the family was obtained, and then the model was utilized to analyze protein sequences. Results indicate that HMM can express particular family of protein, and recognize the given protein sequences of the family from many sequences. PMID- 12557522 TI - [Cloning expression of a novel human protein-disulfide isomerase like cDNA]. AB - Using the large scale sequencing, a novel human cDNA of 1,645 bp was screened from the cDNA library of human fetal brain. The cDNA contains an ORF encoding a 296-aa protein with a calculated molecular weight of 34.0 KD. Compared with that of current sequence databases, the putative protein was found to have 36% homology with protein-disulfide isomerase (PDI). So this cDNA was named PDI like (PDI-L) gene. Multiple tissue Northern blot analysis shows that PDI-L cDNA is expressed in heart, brain, liver, kidneys and so on. The ORF fragment of PDI-L cDNA was inserted into reconstructed pBV220 expression vehicle and its predicted expressive protein was obtained. PMID- 12557523 TI - [Study of the average FR of the MUAP's in a couple of agonist-antagonist muscles]. AB - "Common Drive" is presented recently as a new concept used to explore the control mechanism of neuromuscular system. In this paper, the average firing rate (FR) of the motor unit action potential (MUAP) is estimated by means of decomposition technique for needle electromyographic (NEMG) signals obtained from elbow joint agonist-antagonist muscle pair with constant contraction force. The change tendency and correlation of the average FR with time are studied. The results of the experiment show that, no matter flexion or extension of the elbow joint, the average FR of both motor units(MUs) in the couple of agonist and antagonist descends with time, and the variations of their amplitude and fluctuation are highly correlated. This indicates that when two antagonist muscles are activated simultaneously to stiffen a joint, the nervous system views them as one unit and controls them in similar fashion. It also confirms the existence of "Common Drive" phenomenon at joint level. PMID- 12557524 TI - [Effects of external counterpulsation on the pulsatility of blood pressure in human subjects]. AB - Pulsatile blood flow plays an important role in maintaining normal vascular endothelial function. Quantitative measurement of pulsatility of human arterial blood pressure and the influence of enhanced external counterpulsation (EECP) on the pulsatility were investigated in this study. Eight healthy young male volunteers aged 22 to 35 were included. A 4F tip transducer catheter was inserted under local anaesthesia into the radial artery up to the aortic arch. Intraarterial blood pressure was recorded before and during EECP. Blood pulse pressure, pulsatility index (ratio of peak pressure to end diastolic pressure) and standard deviation of blood pressure in 5 cardiac cycle was calculated to evaluate the pulsatility of arterial blood pressure. The results showed that blood pulse pressure, pulsatility index and standard deviation of blood pressure were elevated from 47 +/- 5 mmHg, 1.64 +/- 0.11 and 13.6 +/- 1.5 mmHg to 77 +/- 3 mmHg, 2.46 +/- 0.25 and 19.3 +/- 2.2 mmHg before and during EECP respectively (P < 0.05). Decreasing of systolic pressure and increasing of diastolic pressure during counterpulsation were also observed. EECP gives an elevation of pulsatility to human blood pressure. PMID- 12557525 TI - [The relationship between obesity, intra-abdominal fat area and bone mineral density and bone strength]. AB - Recently some studies have showed that bone strength may avoid the misdiagnosis and missed diagnosis of osteoporosis(OP) by bone mineral density (BMD) alone. It was found that corrected bone mineral content (cBMC) was well correlated with bone strength. In this study bone strength was replaced by cBMC to compare it with BMD, and evaluate whether BMD has limitations in the diagnosis of OP in obese individuals. one hundred and two healthy subjects in the community with the body mass index (BMI) between 25 and 39 kg/m2 were enrolled. There were 31 males and 71 females with a mean age of 37.2 +/- 9.4 years. Their BMD and BMC of lower limbs, trunk and the whole body were measured with dual energy X-ray absorptiometry (DEXA). cBMC was expressed by BMC divided by weight. Intra abdominal fat area (IAFA) was measured by CT scan at the level between 4th-5th lumbar vertebra. Linear correlation analysis with SPSS 10.0 soft pack was used for statistics. The results showed that there was a positive correlation and tendency between body weight, BMI and BMD values of lower limbs, trunk and the whole body. However, negative correlation and tendency were demonstrated between body weight, BMI and cBMC, between IAFA and BMD of lower limbs, trunk and the whole body and between IAFA and cBMC as well. We found that the tendency of changes of cBMC and bone strength was similar. So we suggest in certain extent that cBMC could replace bone strength in the diagnosis of OP in obese individuals. PMID- 12557526 TI - [The delay time of pediatric LVAD of ECG-triggering mode]. AB - Electrocardiac signal is one of the most important signals which is used to trigger ventricular assist device (VAD), and the delay time of VAD assistance is very important to get a satisfied result. Proper delay will give VAD relatively enough time to assist, avoiding left heart failure caused by the collision of the heart and VAD during systolic phase. This becomes much more important when the left atrium drainage is insufficient. The aim of our study is to set up an equation to calculate the delay time by RR interval. We try to set up an equation about RR and R-Ao like: R-Ao = A x (RR)n + B(A and B are constant). RR represents the RR interval and R-Ao represents the duration of the period between the peak point of QRS and the point of aortic valve closing; First, calculate RR according to weighting average method, and then, calculate the anticipant R-Ao according to the before-mentioned equation. After adjustment, R-Ao will be used as assistance delay time. R-R interval was measured in 457 selected pediatric patients who were undergiong left heart catheterization and who did not have arrhythmias. From the ECG recording during catheterization, R-R interval was measured while R-Ao was obtained from aortic pressure wave chart; Plot graphs with R-Ao as dependent variable and (RR)n as independent variable; find out correlating model and calculate the arguments A and B of R-Ao = A x (RR)n + B. The results showed that the relation between (RR)1/3 and R-Ao is the most significant, the relation coefficient is 0.733, the regress coefficient is -0.182 (P < 0.001) and the interception is 1.070. This means that R-Ao = (-0.182) (RR) 1/3 + 1.070. The likelyhood degrees of different sections differ markedly. When heart rate is less than 120 beats per min. The relation argument is about 0.733 while 0.45 when heart rate is more than 120 beats per min, Therefore, we can use the equation R Ao = (-0.182) (RR)1/3 + 1.070 to calculate R-Ao when heart rate is less than 120 beats per min. PMID- 12557527 TI - [An experimental study on the acquisition of HRV signals with high reliability]. AB - The acquisition of heart rate variability (HRV) signals is very important to physiological research and clinical diagnosis. In order to ensure the accuracy of acquired heart rate variability signals, we must consider the method of acquisition of heart rate variability signals. By using the relation between the signal singularity and its wavelet transforms, a software for R wave in detection is designed. The result is very satisfying in detecting R wave of ECG database of MIT/BIH. PMID- 12557528 TI - [Hemolysis test of the five kinds of impeller blood pumps in vitro]. AB - Hemolysis caused by blood pumps is a very important characteristic. In vitro hemolysis test circuits were constructed to operate the model I centrifugal pump, the model II axial flow pump, the magnetic coupling pump and the model I & II spiral mixed pump. The output of all pumps was set at flow 5 L/min, an average pressure of 100 mmHg. Experiments were conducted for 4 hours at room temperature(25 degrees C) with 500 ml fresh anticoagulant sheep blood. Blood samples were taken for plasma free-hemoglobin measurement, and the change in temperature at the pump outlet port was measured during the experiment. Calculate the normalized index of hemolysis (NIH). The results showed that there was no relationship either between the pump rotational speed and the NIH in five types of blood pumps, or between change in temperature at the pump outlet port and NIH. The dynamic fluid field caused by pump design and structure could be the main cause of blood damage. PMID- 12557529 TI - [Construction and expression of somatostatin (S14) and hepatitis B surface antigen gene in yeast Pichia pastoris]. AB - To construct the ss/HBsAg protein gene-engineering vaccine for developing the diagnosis and cure tumors in clinical medicine and promoting the growth in animal husbandry production. A pair of primers were designed according separately to the sequence of Somatostatin gene(S14) and HBsAg gene. Their gene fragments were separately amplified by using PCR and cloned, following sequencing, the DNA fragments were inserted into pBluescript vector. Then the ss/HBsAg chimera was constructed and was cloned into pPICZaA plasmid, and transformed into electroporated Pichia pastoris. High yield protein expression was obtained. Expressed protein was proved with high specificity and it's molecular weigh was about 28 KD identified by SDS-PAGE and Western blot. PMID- 12557530 TI - [The state and development of cell image segmentation technology]. AB - This paper describes the state and the development of the application of the modern and traditional image segmentation technology in cell slice image segmentation. It includes edge detection, regional segmentation, wavelet transform, fuzzy mathematics, artificial neural networks, morphological image segmentation and so on. At last, the paper summaries that it is difficult to generally segmentate any kind of biological cell slice image automatically because of the complex structure of cell and cell slice image is not even gray distributed. It should be pointed out that general automatic cell slice image segmentation will be achieved only if visual mathematics model corresponding to mammalian vision systems is setup entirely. PMID- 12557531 TI - [The improvement of signal-to-noise ratio in magnetic resonance image]. AB - Digital image processing and magnetic resonance imaging from magnetic resonance image processing technique. This technique will not only promote MRI's research and development, but also bring new challenges for digital image processing. Because the information in the MRI is very important to medical treatment, it is a primary task that how to decrease noise from MRI by use of PC and digital image processing. In this paper, we analyzed the noises from a variety of sources, and introduced all kinds of techniques to reduce these noises. PMID- 12557532 TI - [Review on image reconstruction for positron emission tomography]. AB - Positron emission tomography (PET) is one of the most important applications in nuclear medicine and has become a powerful tool for scientific research. This paper is a symposium of the current status of image reconstruction for PET. The latest advances in the field of filtered-backprojection algorithm and expectation maximization algorithm are also introduced. PMID- 12557533 TI - [Biomechanical mechanism of diffuse axonal injury]. AB - As one type of primary brain injury, diffuse axonal injury (DAI) has specific traumatic mechanisms. The occurence of DAI is close to the loading property, loading manner, structural characteristics of skull, brain tissue and neck. This article demonstrated how the stress and strain varied in brain tissue effected by the load magnitude, load waveform, load frequency, load duration, linear acceleration, rotational acceleration, compounded linear/rotational acceleration, brain tissue, cerebral falx, cerebellar tentorium, skull and neck, and what are the relationships between these factors and the event of DAI. PMID- 12557534 TI - [The application and development of artificial intelligence in medical diagnosis systems]. AB - This paper has reviewed the development of artificial intelligence in medical practice and medical diagnostic expert systems, and has summarized the application of artificial neural network. It explains that a source of difficulty in medical diagnostic system is the co-existence of multiple diseases--the potentially inter-related diseases. However, the difficulty of image expert systems is inherent in high-level vision. And it increases the complexity of expert system in medical image. At last, the prospect for the development of artificial intelligence in medical image expert systems is made. PMID- 12557535 TI - [Biology and pathobiology of death receptors in liver]. AB - Death receptors induce apoptosis by intracellular signaling transmission and relate to body growth, development, disease and death. This paper reviews the structural character of death receptors, and the mechanisms by which apoptosis is induced. The biology and pathobiology of these death receptors in liver are discussed as well. PMID- 12557536 TI - [The study of a image compression method based on wavelet statistical characteristic]. AB - The statistical characteristic is the key to image compression. Here we have studied the statistical characteristic of wavelet subimage, and designed a new image quantization algorithm referred to statistical characteristics of both wavelet subimage and human visual. PMID- 12557537 TI - [Improvement on simulation algorithm of excitation propagation in heart modeling]. AB - It is important to simulate the excitation propagation process of cardiac bio electricity in the research of ECG forward problem. Traditional methods describe them with wave simulation algorithm such as LFX simulation algorithm and vector propagation algorithm etc, these methods have some problems to certain extent, due to the presence of discreteness of space and time and asymmetry of the myocardium. This paper discussed the simulation algorithm in 2-dimension space under the circumstance of layered and non-layered structure of myocardium. By calculating the theoretic values of simulating time based on Huygen's principle, we found that there were errors in LFX algorithm and no errors in vector propagation algorithm under the circumstance of non-layered structure of the myocardium, no mater what myocardium is isotropic or anisotropic. However, there exist errors from both algorithms when the myocardium has the layered structure. An improved algorithm is proposed and the simulations have been performed to examine the efficacy of the new algorithm, and the errors are reduced obviously. By increasing the number of myocardial blocks in the model, we also analyzed its influence on the error of simulation algorithm. PMID- 12557538 TI - [The development and clinical application of acoustic diagnostic technique in hip joint]. AB - This study aimed to develop a non-invasive diagnostic technique for the measurement of acoustic transmission in hip joints to reflect the dynamic response of the observed structure. This instrument consists of three sub systems. An stimulation system applied a vibratory force at the sacrum of the test subject. A transduction system included a pair of identical microphones was installed in the tubes of two stethoscopes, which were placed at the greater trochanters on both sides for picking up the acoustic signals transmitting across the hip joints. The data acquisition and analysis system was a portable frequency analyser with a program of dual channel digital filter for measuring the power of acoustic signals in 1/3-octoscn frequency bands. Twenty-seven normal adults, 20 normal pre-school children and 40 normal neonates were recruited for the testing. Coherence function (CF) of the signal and the discrepancy (D) of bilateral hips were measured during the testing. The results from the three groups showed that there was a high coherence of the signals (CF > 0.9) and a small discrepancy (D < 3 dB) between bilateral hips in the frequency range of 200-315 Hz. For normal neonates, the frequency range (160-315 Hz) was wider in which the acoustic signals maintained a high coherence (CF > 0.94) and a smaller discrepancy (D < 2 dB) were observed between bilateral hips. This study has shown that the development of the acoustical diagnostic technique could provide a practical method with objective parameters. The results obtained in this study can offer a base for further investigation of hip disorders, particularly those related to structural abnormalities of hip joints. PMID- 12557539 TI - [The effect of HJ-1 NO--HFJV respirator on treating pulmonary edema caused by seawater drowning]. AB - The study was to observe the therapeutic effect of HJ-1 NO--HFJV respirator on treating pulmonary edema caused by seawater drowning. Seawater was infused into the rabbit's lung to establish the animal model of pulmonary edema caused by seawater drowning(PE-SWD). Then the animals were divided into three groups: simple PE-SWD model as control group, treat group(animal model treated with HFJV respirator and four medicines) and HFJV respiratior plus NO group. Pao2, Sao2 and pH were measured by the blood-gas analyzer. The survival time and seawater drowing-respiratiory distress syndrom(SW-RDS) were observed. The results showed that Pao2, Sao2 in NO group were remarkably higher than that of PE-SWD control group, and the survival time was longer than that of medicine treated group and the incidence of SW-RDS decreased to zero. We assume that HJ-1 NO-HFJV respirator is efficient on treating pulmonary edema. PMID- 12557540 TI - [Study on multi-pole RF ablation system]. AB - Radio frequency ablation has become a valuable method in treating cancers or tumors for its wide adaptability, better efficacy, convenience and safety. This paper introduces the research and development of a multipolar RF tumor therapy system based on the technology of destroying hypoxic cell. This is an intellectualized tumor curing system. PMID- 12557541 TI - [A 3D real-time visualization system of medical image]. AB - Based on the integration of the advanced visualization toolkit (VTK) and the real time rendering System (VolumePro), 4D View, a 3D real-time visualization system of medical image, which applies real-time 3D medical image (such as MR, CT image, etc.) visualization and interaction was developed. Firstly the VTK, VolumePro and their integration were introduced briefly; then the system design and function template of 4D View system was discussed in detail; finally some visualization results acquired through 4D View was illustrated. According to the results, 4D View system can effectively resolve the poor real-time characteristic of 3D medical image visualization and interaction, so it will have a wide application area of the clinical diagnosis, therapy and medical research, etc in future. PMID- 12557543 TI - [Fusion expression of PLA2 gene from Lapemis hardwickii in E. coli]. AB - The gene encoding PLA2(PLA2-9) from Lapemis hardwickii Gray venom was cloned to the 3' and of the thioredoxin gene (HP-trxA and trxA) in plasmid pthioHisC and pTRX to construct the pThioHisC-PLA2, and pTRX-PLA2 fusion expression vector. The fusion protein of PLA2 can be expressed in the two different systems induced by IPTG at 25 degrees C, but the expression level and the solubility of the fusion protein in pTRX were better than that in pThioHisC. The expressed product in the two systems were purified by immobilized metal-chelate affinity chromatography. The Trx-PLA2 fusion protein with over 85% purity was obtained and HP-Trx-PLA2 fusion protein can not be purified since it dose not exhibit affinity to the medium. So, the pTRX-PLA2 vector system was established for the large-scale expression and purification. PMID- 12557542 TI - [Cloning and sequence analysis of aspartokinase genes from Corynebacterium crenatum]. AB - Aspartokinase genes (ask) from wild-type Corynebacterium crenatum AS1.542 and an AEC-resistant mutant Corynebacterium crenatum CD945 were cloned and sequenced. Analysis of ask sequence shows a exchange in a single base pair at position 1199 from T to C, leading to an amino acid change in the beta subunit of Aspartokinase. Leu80 in the wild-type is converted to Pro80 in the feedback resistant enzyme. The substitution is located in ACT domain, a region regulated by concentration of lysine. The ORF sequence of ask from C. crenatum AS 1.542 shows homologies of 97.23%, 97.55% and 97.62% to those from C. glutamicum, C. flavum and B. lactofermentum. And the amino acid sequence deduced from ORF displays homologies of 99.76%, 99.52% and 99.76%, respectively. But there is much variation in the upstream sequence of C. crenatum AS 1.542 ask promoter compared to those from other Corynebacteria. PMID- 12557544 TI - [Molecular cloning and amino acid composition analysis of a halophilic thiolase gene]. AB - 5' and 3' end sequence of acaBl gene as primers, the gene of halophilic thiolase from haloarchae, Halobacterium sp. ZP-6 was cloned and its amino acid composition was calculated. Compared with non-halophilic thiolase, the halophilic thiolase contains more negative charge amino acid, less positive amino acid and less strong hydrophobic amino acid, and use preferably small side-chain amino acid. Those suggest that electrostatic screen, hydrophobic effect and surface tension all contribute to halophilic properties of thiolase. PMID- 12557545 TI - [Cloning and expression of VEGF receptor Flt-1 gene in S. lividans TK24]. AB - Using RNA extracted from human umbilical vein endothelium cell as a template, the gene VEGF receptor Flt-1 was amplified by RT-PCR. Recombinant plasmid pSGLgpp-F was constructed and was transformed into S. lividans TK24. With the detection of SDS-PAGE and Western blot, a specific band being same to the reports near 63.6 kd was found. The results showed sFlt-1 was successfully expressed in S. lividans. The result of the binding assay of receptor-ligand for sFlt-1 showed sFlt-1 has the biological activity of binding with its ligand VEGF. PMID- 12557546 TI - [Study on effect of Vitreoscilla hemoglobin gene expression on growth and metabolism of Streptomyces aureofaciens]. AB - Vitreoscilla hemoglobin gene was expressed in S. aureofaciens through promoter of tetracycline resistance gene. Characteristics of S. aureofaciens growth and metabolism were studied in 1 m3 fermenter. In high dissolved oxygen concentrations, expression of hemoglobin gene had little effect on growth and metabolism of S. aureofaciens, and there were no obvious differences between engineering strain and control. The chlortetracycline of engineering strain was 22905 u/mL and of control was 22896 u/mL respectively. Under conditions of low dissolved oxygen, expression of hemoglobin enhanced growth, maintenance of energetic mycelium configuration and chlortetracycline yield of S. aureofaciens: the mycelium concentrations of engineering strain were more about 5%-10% and yield was more 11.4% than control. PMID- 12557547 TI - [Improvement of recombinant xylanase XynF1 from Aspergillus oryzae expressed in Pichia pastoris by site-directed mutagenesis]. AB - The recombinant xylanase XynF1 from Aspergillus oryzae expressed in the methylotrophic yeast Pichia pastoris was improved by site-directed mutagenesis. The mutant I156A secreted about 47.7 U/mL xylanase XynF1 in the BMMY medium. The mutant xylanase XynF1 was purified by ion exchange and gel filtration chromatographies. The molecular weight of purified XynF1 was 35 kD. XynF1 was stable between pH 4-9 and its optimum pH was 7.0. The optimum temperature of XynF1 was 45 degrees C and it was stable below 50 degrees C. The properties of mutant were same as the parent's except the stability of temperature. PMID- 12557548 TI - [Construction of a set of secreting expression vectors for Saccharomyces cerevisiea]. AB - The DNA fragment ecoding the Signal peptide of inulinase of Kluyveromyces smarxianu was synthesized chemically. This fragment was cloned in-frame in the expression vector pYES2 of Saccharomyces cerevisiae, resulting in a set of new secreting expression vectors pYES2 I, pYES2 II, pYES2 III. The L-Asparaginase gene (ASN) of E. coli and alpha-acetylactate decarboxylase gene (ALDC) of B. brevis which were amplified by PCR and cloned into the new vectors respectively were transformed into Saccharomyces cerevisia, and most of enzyme activities were secreted into the medium. The new secreting expression vectors still have excellent segregational stability even after growth for 100 h in the absence of selective pressure. PMID- 12557549 TI - [Gene cloning and sequencing of chicken anemia virus(CAV) isolated from Harbin]. AB - A Chicken anemia virus has been isolated from a chicken flock in Harbin of China. The genome of the ivrus was cloned through polymerase chain reaction(PCR) and sequence of the genome was analyzed. The cycle genome is made of 2298 base pairs including three overlapping open reading frames(vp1, vp2, vp3) and a regulative region. Comparing sequence of the genome through BLAST in GenBank, this sequence exhibits 96.9% identity with other genome of CA Vs and least. Multiple alignment of this genome of this virus, 26p4, strain isolated in Germany, strain isolated in Malaysia and Cux-1 found that this sequence exhibits 98.2% (42/2298), 98.2% (42/2298), 96.9% (72/2298) and 97.5% (60/2319) identify with them, respectively. A new CAV strain was isolated and it has better identify with CAV isolated in Europe countries than is Asia country Malaysia. Multiple alignment of VP1, VP2, VP3 of 26p4, strain isolated in Germany, strain isolated in Malaysia, Cux-1 and strain isolated in Harbin of China found the VP2 the most conservative. PMID- 12557550 TI - [Recombinant fowlpox virus expressing HA from subtype H9N2 of avian influenza virus and its protective immunity against homologous challenge in chickens]. AB - The hemagglutinin (HA) gene from the AIV, A/Chicken/China/F/1998(H9N2), was amplified with the RT-PCR technique and directionally inserted into transferring vector 1175, resulted in recombinant transferring vector 1175HA. In order to generate recombinant fowlpox virus expressing HA(rFPV-HA), the recombinant transferring vector 1175HA was used to transfect the chicken embryo fibroblasts(CEF) pre-infected with wide type fowlpox virus. Then, by selection of blue plaques on the CEF overlaid with agar containing X-gal, rFPV-HA was obtained and purified. The expression of HA by rFPV-HA was detected in the recombinant virus-infected CEF by indirect immunofluorescence. Experiments on chickens demonstrated that rFPV-HA could induce detectable HI antibodies 7 days post vaccination and those HI antibodies of relatively high titers could persist 55 days. rFPV-HA also had the same protective efficacies to suppress SPF chickens or commercial broiler chickens with antibodies against FPV from shedding challenged virus from intestine as inactivated vaccine in oil emulsion. PMID- 12557551 TI - [Cloning and prokaryotic expression of viral binding protein(VBP) gene from endosybiotic bacterium of Rhopalosiphum padi]. AB - Viral binding protein gene from Rhopalosiphum padi Yangling biotype was amplified by PCR method and then cloned. The complete nucleotide sequence of the gene was determined, It has 1647 nucleotides encoding 548 amino acids. Comparison showed this gene has 97% identity on nucleotide level with Buchnera groEL-AM gene of Rhopalosiphum padi American biotype, while has 97.4% identity on amino acid level was found between this two genes. The VBP gene was ligated into pBV221 and pET30a expression vector and expressed the aim protein 63 kD and 69 kD. PMID- 12557552 TI - [Production of monoclonal antibodies to tomato mosaic virus and application in virus detection]. AB - Four hybridoma cell lines secreting monoclonal antibodies(MAbs) against Tomato mosaic virus(ToMV) were produced by fusing mouse myeloma cells(SP2/0) with spleen cells from BAL B/c immunized by the ToMV particle. The four MAbs could specifically react with ToMV, and the MAbs from two cell lines can react with ToMV and TMV simultaneously. The titres of ascitic fluids of four MAbs ranged from 1:32,000 to 1:1,024,000 with ELISA, and the sensitivity for detection virus from the plant sap reached over 1:2000 dilution. The MAbs didn't crossreact with other plant viruses. The result of Western-blot showed that two MAbs can react with the 17.6 kD ToMV coat protein submit specifically, while the other two MAbs can not react with it, they are supposed to against conformational determinants of ToMV CP. PMID- 12557553 TI - [Site-directed construction of in-frame deletion mutants of Corynebacterium glutamicum]. AB - This study is focused on Corynebacterium glutamicum, an important producer in amino-acids industry, by the protocol with construction of the in-frame deletion fragments of targeting genes, application of DNA recombination technique and so on. We achieved successfully the defined deletion mutants in frame of the single gene or double genes. The in-frame deletion mutants are subsequently analyzed and confirmed by polymerase chain reaction and sequencing. PMID- 12557554 TI - [Type III secretion system in Se9R can recognize and secret harpin in Erwinia amylovora]. AB - Bacterial pathogens of both animals and plants use type III secretion machines to secret virulence proteins. The signal that leads to tye type III secretion is encoded as stem loop structure in the messenger RNA. Harpin, produced by Erwinia anylovora is secreted via type III secretion. Erwinia carotovora Se9R and Erwinia amylovora belong to the same genus and they use type III secretion machines to secret virulence proteins. We used PCR to amplify hrpN from pCPP430 which harbors hrpN gene cluster, and cloned it to pGEM-T vector to get pWGF1, which was then chemically transformed into DH5 alpha (pCPP430hrpN-) and electrotransformed into Se9R. DH5 alpha (pCPP430hrpN-/pWGF1) induced hypersensitive response on tomato leaf and Se9R(pWGF1) showed significantly reduced virulence than Se9R on Chinese cabbage leaf. Western-blotting analysis of the CFEP of Se9R(pWGF1) showed production of harpin protein. These results suggest the type III secretion machine in Se9R can recognize secretion singal in the gene of harpin in Erwinia amylovora and secret it as a biologically native form. PMID- 12557555 TI - [Purification and properties of an endocellulase from the thermophilic fungus Chaetomium thermophile]. AB - An endocellulase from culture supernatant of a thermophilic fungus Chaetomium thermophile was purifided to homogeneity, by using ammonium sulfate fraction, DEAE-Sepharose Fast-flow chromatography, Phenyl-Sepharose Fast Flow chromatography and Sephacryl S-100 chromatography. The enzyme was a glycoprotein with an apparent molecular weight of 67,800 and 69,800, as determinded by 12.5% SDS-PAGE and gel filtration respectively. The endocellulase was optimally active at pH 4.0-4.5 and 60 degrees C. It was thermostable at 60 degrees C and retained 30% activity after 60 min at 70 degrees C. The half life time of the enzyme at 80 degrees C was 25 min. Different metal ions showed different effects on the endocellulase activity. Na+ enhanced the enzyme activity, whereas Fe2+, Ag+, Cu2+, Ba2+ and Zn2+ cause obvious inhibition. But it didn't work on crystalline cellulose. PMID- 12557556 TI - [Community constitute and phylogenetic analysis on soil uncultured microorganism]. AB - Many evidences show bachteria that can be cultured in lab are only the minority of germs existing in nature. Most of bacteria, containing rich valuable gene resource, are unknown for they are uncultured. To research these uncultured bacteria, Directly extract crude DNA from farm soil, separate 16S rDNA mixture amplified by PCR from the crude DNA, sequence it and compare the sequences with the records of NCBI to classify the germs. The significant microbe community difference from respective soil, which have possibility in developing a new tool for researching soil bacteria diversity were finded. PMID- 12557557 TI - [Asymmetric microbial reduction of organosilyl ketone with immobilized Saccharomyces cerevisiae cells]. AB - Asymmetric microbial reduction of acetyltrimethylsilane with immobilized Saccharomyces cerevisiae cells was studied. The effects of shake speeds, substrate concentration, immobilized cell concentration, pH and reaction temperature on the initial reaction rate, maximum yield and optical purity of the product were systematically studied. All above mentioned parameters have important effects on the raction. The optimum shake speed, substrate concentration, immobilized cell concentration, pH and reaction temperature were found to be 150 r/min, 14 mmol/L, 0.15 g/mL, 7.3 and 25 degrees C-30 degrees C respectively for the reaction, under which the maximum yield and optical purity of the product were as high as 84.9% and 90.2% ee. PMID- 12557558 TI - [Study on Pseudomonas sp. WBC-3 capable of complete degradation of methylparathion]. AB - A bacterial strain was isolated from the polluted soil around Shashi Pesticides Factory, which was capable of complete degradation of methylparathion. Through chemotaxonomic characterizations and phylogenetic inference based on 16S rDNA sequence analysis, the strain was identified as a member of the genus and was named as Pseudomonas sp. WBC-3. It can tolerate high-concentration methylparathion up to 800 mg/L in basic medium and up to 2000 mg/L in 0.1% glucose medium. Using 300 mg/L methylparathion as its sole carbon and nitrogen sources, the strain was able to degrade 15 mg of parathion per liter per hour and reached its stationary phase in about 22 hours. The strain possessed broad spectrum degradative capability to kinds of organophosphorus pesticides and aromatic compounds. Its organophosphate hydrolase was purified from the periplasm of WBC-3 to homogeneity, through a whose process consisting of ammonium sulfate precipitation, Sephadex G-75 gel filtration, Q Sepharose FF ionexchange column chromatography. The purified enzyme showed a single band on SDS-PAGE gel with an approximate molecular weight of 33.5 kD. PMID- 12557559 TI - [Studies on production of docosahexaenoic acid by Thraustochytrium aureum]. AB - The effects of carbon source, Na-glutamate, salinity, and incubation temperature on the growth of Thraustochytrium aureum H0 and the production of docosahexaenoic acid were studies. The time course of the cell growth of Thraustochytrium aureum H0 was determined. The results show that glucose is the optimum carbon source and the optimum concentrations of glucose and Na-glutamate are 30 g/L and 5 g/L, respectively. The suitable salinity is half of salinity of sea water. Under the optimum culture conditions, 6.2 g/L of the dry cell weight and 0.51 g/L of the docosahexaenoic acid were obtained. PMID- 12557560 TI - [Halocin: protein antibiotics produced by extremely halophilic archaea]. PMID- 12557561 TI - [Structural feature of bacterial cell surface S-layers and its functions and application potential]. PMID- 12557562 TI - [Research progress on the glycerol kinase]. PMID- 12557563 TI - [Advance in studies on Cyt proteins of Bacillus thuringiensis]. PMID- 12557564 TI - A potentially dangerous flaw in the Mississippi mental health commitment process. PMID- 12557566 TI - I am following a health care professional who, I think, has a drug problem. PMID- 12557565 TI - Hunting injuries in Mississippi. AB - It is clear from the data that hunting injuries in Mississippi mirror those which occur nationwide. The vast majority of these injuries are easily preventable by following common sense hunting safety rules. As with firearm injuries, tree stand injuries are clearly preventable by following common sense rules (Table 1). Most falls are due to poorly constructed or old wooden tree stands that come apart or become detached from the tree thereby causing the hunter to fall. All bolts and fasteners should be inspected and replaced if found to be defective prior to use of the stand. Old, broken or rotten lumber should be replaced. Tree stands should only be placed in healthy, mature trees with strong healthy limbs. Unfortunately, tree stand hunters seldom wear safety harnesses when hunting. These devices, when used properly (worm around the chest under both arms), can greatly decrease morbidity and mortality associated with falls. To minimize trauma, tree stands should be constructed no greater than 20 feet above ground level. Hunters should be cautioned to wear non-slip boots [table| see text] and to apply a non-slip covering to the floor of their tree stands prior to use of the stands. Other safety tips include the removal of all logs, stones or other obstructions from around the base of the tree. As with all hunters, tree stand hunters are advised to carry a compass, whistle, flashlight and cellular phone with them during their hunt. Hunting equipment, including bows and arrows and all firearms should never be carried up to the stand. This equipment should only be lifted or lowered to or from the stand with the use of a haul line. Hunters must remember the 10 commandments of gun safety (Table 2). Every firearm should be treated as if it were loaded. Firearms should not be pointed at anything other than the intended target and the muzzle must be controlled at all times. The action and barrel should be inspected prior to loading ammunition to ensure that they are clear of obstructions. The target must be clearly identified before discharging the firearm. All firearms must be kept separate from their ammunition and unloaded when not in use. Alcohol and any mind-altering substances (antihistamines, etc) must be strictly avoided when hunting. Hunters should avoid shooting at hard, flat surfaces in an attempt to prevent ricochets, should never pull a loaded gun toward themselves, and should never climb over a fence or other obstacle with a loaded firearm. Additional common sense safety precautions include the use of the "gun safety." The safety should be "on" at all times and switched off only when the rifle is sighted in, on the game just before discharge. This simple precaution may well prevent many of the accidental shootings that occur when firearms are dropped. When firearms are dropped, the hunter must carefully pick up the gun butt first while leaving the muzzle pointed toward the ground. Rifles should be left unloaded until game is sighted and should be loaded only in the immediate anticipation of discharging the firearm at game. Hunters are also advised to hunt in-groups and should notify family members or friends of the approximate time they anticipate entering and leaving the hunting area. Bright orange hunting vests have been shown to greatly increase the hunter's visibility and should always be worn when in the field during hunting season. Hunters are also advised to layer their clothing to prevent cold injuries. A backpack containing a flashlight, compass, prescription medications, first aid kit, cellular phone and high calorie foods should be taken into the field with the hunter. Bow hunters can drastically reduce injuries and death by following a few common sense rules (Table 3). All arrows should remain sheathed until prey is sighted at which time the arrow may be nocked to the bowstring. Bows should never be drawn until the hunter is ready to fire at the target. At all times, the bow and nocked arrow should be pointed in a safe direction. Reminding hunters of the major causes of accidents as well as how to prevent them will help to decrease hunting injury and death rates in Mississippi. Careful planning and safety precautions on the part of all hunters are essential to their own safety as well as that of others. PMID- 12557567 TI - "Dr. Shug". PMID- 12557569 TI - Tobacco Quitline Mississippi 1-800-244-9100. PMID- 12557570 TI - [Integration of metabolic data by positron emission tomography in image-guided neurosurgical interventions]. AB - The potential of positron emission tomography (PET) to investigate accurately in vivo pathophysiology has opened new perspectives for the management of neurological disorders, especially brain tumors. Integration of PET in the management of brain tumors represents a unique way to provide metabolic and functional information that can be used for diagnosis, guidance of therapies, and treatment monitoring. We have developed a stereotactic method allowing the routine integration of PET images in neurosurgical procedures with a high precision. Our ten year experience with the use of PET in stereotactic conditions for neurosurgical guidance, indicates that PET integration in image-guide neurosurgical approaches, including [1] stereotactic biopsy, [2] neuronavigation and [3] radiosurgery with Leksell Gamma Knife, is feasible and worthwhile, in that it provides complementary information that contribute to the optimization of the neurosurgical treatment of brain tumors. PMID- 12557571 TI - [Regeneration of ciliated cells in the internal ear]. AB - Hair cells are the mechanotranducer transforming the sound into a bioelectrical signal. Hair cell and supporting cell productions are completed during early embryonic development of the mammalian cochlea. In mammalian, after an injury, no hair cell replacement is observed, as opposed to birds, where regenerative mechanisms produce new sensory cells and restore the auditory function. However, a production of hair cells occurs in the mammalian sensory epithelium. Progenitor cells, isolated from newborn rats, proliferate and differentiate in hair cells and supporting cells. Supernumerary hair cells also arise in the cultured organ of Corti. This model is used to investigate the role of cell cycle regulator molecules and cell-cell interaction. The persistence of sensory cell progenitors in adult mammalian organ of Corti and the understanding of the mechanisms leading to the production of hair cells, in the developing cochlea, open the prospect of hair cell regeneration in the mature inner ear. PMID- 12557568 TI - Looking back: the president's address in 1917: "The ethics of medicine are but the ideals of gentlemen". PMID- 12557572 TI - [Academic eulogy for Professor J. Beumer]. PMID- 12557573 TI - [Heat shock proteins, embryogenesis and evolution]. AB - We present results about immunohistochemical identification of several heat shock proteins (HSP'S) during mouse normal and teratological embryogenesis. Apoptotic cells express very specifically and precociously HSP 110. This fact permits to identify apoptotic cells before apparition of morphologic features of apoptosis, but also to quantify the process of cell death in some teratological models, particularly administration of retinoic acid. HSP 86 is expressed in some cell populations, and particularly permanent in germ cells. Our observations brought us to discuss the potential protective role of HSP on germ cells, and the consequence of their inactivation in the macroevolution process, as well as the role of apoptosis in teratology. PMID- 12557574 TI - [Evolutionary strategy of antibiotic resistance]. AB - Antibiotics have reduced the mortality from infectious diseases, but not the prevalence of these diseases. Use, and often abuse, of antimicrobial agents encourages the evolution of bacteria toward resistance, often resulting in therapeutic failure. This evolution is due to the emergence of "new" resistance mechanisms and to the spread of well-characterized mechanisms of resistance, to the majority of bacterial species. Bacterial resistance can be intrinsic or acquired. Intrinsic resistance is species or genus specific and delineates the spectrum of activity of the antibiotic. Acquired resistance is present in only certain strains of a species or of a genus. The latter results from mutation in a gene located in the host chromosome, or a plasmid, or from acquisition of new genetic information by a bacterium mainly by conjugation or transformation. PMID- 12557575 TI - [Role of PAI-1 plasminogen activator inhibitor in tumor invasion and angiogenesis]. AB - The plasminogen/plasmin system plays a key role in cancer progression, presumably via mediating extracellular matrix degradation and tumor cell migration. High levels of components of the plasminogen activation system, and paradoxically also its inhibitor, plasminogen activator inhibitor 1 (PAI-1), have been correlated with a poor prognosis for patients with cancers of different types. Recent findings clearly suggest that PAI-1 is essential for capillary sprouting during tumor angiogenesis. Moreover, there is an accumulating evidence that both the urokinase receptor and PAI-1 are multifunctional proteins involved not only in extracellular matrix proteolysis, but also in cellular adhesion and migration through their binding site for vitronectin. The understanding of whether PAI-1 plays a regulatory role in angiogenesis by tightly controlling proteolytic activity, or by influencing cell migration, could allow a new anti-angiogenic approach for tumor therapy. PMID- 12557576 TI - [Helmholtz, a light for the neurosciences]. AB - Helmholtz was an outstanding scientist, and dominated physiology during the second half of the XIXth century in the field of visual and auditory perception. Philosopher, he was deeply indebted to Kant for elaboration of empirical and experimental approach of facts. Man of culture, he laid the foundations of a scientific approach of music and painting. His work remains a fundamental contribution to the theory of knowledge. PMID- 12557577 TI - [Experimental study of systemic ventriculo-arterial coupling. Effects of modifications of thoracic aortic mechanical properties and myocardial ischemia on left ventricular performance]. AB - We quantified systemic ventriculo-arterial coupling in the pressure-volume (PV) plane, using a model of elastic chambers of the cardio-vascular system. In such a model, the ratio between left ventricular (LV) end-systolic elastance, and effective arterial elastance measures the coupling between the heart and the systemic vasculature, and the effects of cardiac diseases and of vasoactive agents on the coupling relationship can be independently evaluated in vivo. Furthermore, the analysis in the PV plane gives insights into LV mechanical efficiency, defined by the ratio between stroke work and pressure-volume area. Our results demonstrating that an augmentation of aortic compliance decreases the energetic cost of LV ejection should be considered when replacing the thoracic aorta by an artificial prosthesis. The homeometric autoregulation, suggesting a mechanism by which the heart can maintain a constant stroke volume, in the face of increased afterload, without using the Starling mechanism and independently of baroreflex integrity, is of paramount importance in heart transplant patients. Indeed, such patients, although lacking cardiac innervation, can adapt their cardiac output, stroke volume, end-systolic and end-diastolic pressures, without simultaneous changes in heart rate. Finally, our data demonstrate that the acutely ischemic LV must face an hostile hemodynamic environment, characterised by early vasoconstriction, later combined with decreased vascular compliance. These results are important when choosing vasoactive agents in the setting of acute myocardial infarction. PMID- 12557578 TI - [Incidence of parasitic diseases in the Russian Federation and basic lines of activities in its stabilization]. PMID- 12557579 TI - [Comparative characterization of a risk for infection with tick-borne encephalitis in the foci with different regional confinement]. AB - A comparative study of the foci of tick-borne encephalitis (TBE) has revealed that the infectivity of vectors taiga ticks (Ixodes persulcatus) plays the leading role in the formation of foci with the highest epidemic activity (including forest steppe ones). The morbidity rates in each regional focus are directly related to the number of tick-bitten persons and to the quantity of the vector. PMID- 12557580 TI - [Epidemiological aspects of leptospirosis in the eastern regions of the Russian Federation]. AB - There have recently substantial changes in the basic parameters of Leptospira infection in the eastern regions. The infection is characterized by a pronounced and steady-state rise in morbidity rates, by its trend for urbanization, by the spread of the disease in recreational areas, by a primary role of dogs as sources of infection, by polymorphism and atypical nature of the clinical picture of Canicola leptospirosis, which make its diagnosis difficult. The anthropurgic foci where dogs and pigs are the main sources of pathogenic leptospiras are of the leading epidemiological significance. The optimum package of measures reducing a risk of human infection is recommended. PMID- 12557581 TI - [Clonorchiasis in the Upper Amur region: biology, epidemiology, clinical presentation]. AB - Results of studies of clonorchiasis in the region of the Upper Amur and its tributaries are presented. Biological features of the pathogen and its epidemiology are outlined. Clinical symptoms were studied in 112 patients. In case of death, a pathoanatomic picture is given. PMID- 12557582 TI - [The duration of action of Bacillus thuringiensis spp. israelensis and Bacillus sphaericus after encapsulation by infusoria Tetrahymena pyriformis]. AB - The study of the duration of the insecticidal activity of a mixture of infusoria with each of these bacteria revealed differences in the pattern of their relations during their long contact. T. pyriformis may be used for the prolongation of its insecticidal action if Bti is applied. An increase in the duration of this action depends on the viability of tetrahymena in the experimental flasks and on the presence of free spores and crystals of Bti. The prolongation of the action may be associated with the multiplication of this type of bacteria in the digestive vacuoles of infusoria. The use of Bsph in the mixture with T. pyriformis caused no increase in the insecticidal activity of bacteria. Tetrahymena increased the duration of Bsph and even reduced the period of its larvicidal activity. Tetrahymena used a sporocrystalline complex of Bsph as food and reduced its water concentration. PMID- 12557583 TI - [Mixed infection in ticks: a rule or an exception?]. AB - A hypothesis for the independence of acts of infection in ticks with pathogens from each other in mixed infection has been tested on the basis of the data available in the literature. In most cases the number of ticks with mixed infection is consistent with the independence hypothesis and fails to reinforce the statement that there is antagonism of pathogens. But in some reports the number of mixed-infected ticks is 3-13 times higher than the calculated number according to the accepted hypothesis. These facts suggest that the natural tolerance of ticks to pathogens due to primary infection may be reduced. Other possible reasons for differences in the actual and calculated number of mixed infected ticks are also discussed. PMID- 12557584 TI - [Impaired intestinal absorption in patients with psoriasis concurrent with chronic opisthorchiasis]. AB - The study was undertaken to characterize small intestinal absorption in patients with psoriasis concurrent with chronic opisthorchiasis. Sixty patients with psoriasis concurrent with chronic opisthorchiasis, 45 patients with psoriasis without helminthiasis, and 15 healthy individuals were examined. The absorptive function of the small intestine was studied by the Kamer method (fat absorption) and with the D-xylose (5 g) test (carbohydrate absorption). Impaired small intestinal absorption of fats and carbohydrates was found in patients with psoriasis concurrent with chronic opisthorchiasis. There was a relationship of these disorders to the clinical picture of psoriasis (stages, duration, skin damage areas, severity). PMID- 12557585 TI - [Parameters of phagocytic activity in patients with toxocariasis]. PMID- 12557586 TI - [Glass slide smears and imprints from infected organs for identification of Leishmania major by polymerase chain reaction methods]. AB - The paper presents data showing that the DNA isolated from the smears and imprints of L. major-infected hamsters is suitable for use in the polymerase chain reaction (PCR) to detect the causative agent of leishmaniasis. The most solid data have been obtained with the smears unexposed to staining and examination by using immersion oil and to benzene treatment. The DNA isolated from these smears infected may preserve for at least 1.5 months in a domestic refrigerator. The immersion oil-treated smears may be also used to identify leishmanias, but DNA should be isolated from the infected specimens of these smears just before PCR. The original primer pair L-unit/L-mail that has shown itself well in the experiments on cultured promastigotes may be, if required, used to differentiate L. major and L. turanica in the infected material collected from infected rodents. PMID- 12557587 TI - [Efficiency of immune reflex diagnosis in the evaluation of some immunologic parameters in subjects with intestinal parasitic invasion]. PMID- 12557588 TI - [Clinical features of giardiasis in children and efficacy of tiberal used for their treatment]. PMID- 12557589 TI - [Experience in treating cysticercosis of the brain with azinox]. AB - A case of a patient resident from Tuva who had cerebral cysticercosis is presented. The diagnosis was established on the basis of the available clinical symptoms, brain RCT and NMRI findings and a positive serological reaction (enzyme immunoassay) with C. cellulosus antigen. The patient received 2 courses of treatment with the Russian drug azinox (praziquantel) in a daily dose of 50 mg/kg body weight thrice before meal. Prednisolone was additionally given in a daily dose of 30 mg. The course of treatment was 14 days. The first treatment days were marked by worse headache. Thereafter treatment induced no complications. Better condition, less headache, lower serological reaction titers, regressed cerebral structural changes were observed after the treatment. RCT and NMRI revealed transformation of cysts to high-density foci and calcificates. PMID- 12557590 TI - [Tasks of the state sanitary-and-epidemiological service of the Ul'ianovsk Region in preventing natural and focal infectious diseases in the light of implementation of the Russian Federation's Law "On sanitary and epidemiological wellbeing of the population"]. PMID- 12557591 TI - [A case of dioctophymosis (Dioctophyme renale) in a girl from Arkhangelsk]. PMID- 12557592 TI - [An outbreak of trichinosis in the Kaliningrad region]. PMID- 12557593 TI - [Incidence of dirofilariasis in the city of Tiumen]. PMID- 12557594 TI - [The range of pathogen variation due to discussion of different hypotheses of plaque]. AB - The paper reviews the hypotheses that explain the mechanism of plague enzooty in natural foci, which are based on a concept of a wide range of plague microbial variability. A comparative analysis of the parameters of variability in the experimentally obtained plague microbial strains and "atypical" natural isolates of the causative agent has led to the conclusion that the mechanism of adaptive variability is due to a phenotypic change in ontogenesis that reflects the philogenetic pathway of the adaptability of a plague microbe to constantly changing living conditions in the ecological niche assimilated by the causative agent. PMID- 12557595 TI - How children and adolescents evaluate gender and racial exclusion. AB - Children's and adolescents' social reasoning about exclusion was assessed in three different social contexts. Participants (N = 294) at three ages, 10 years (4th grade), 13.7 years (7th grade), and 16.2 years (10th grade), fairly evenly divided by gender, from four ethnic groups, European-American (n = 109), African American (n = 96), and a combined sample of Asian-American and Latin-American participants (n = 89) were interviewed regarding their social reasoning about exclusion based on group membership, gender, and race. The contexts for exclusion were friendship, peer, and school. Significant patterns of reasoning about exclusion were found for the context, the target (gender or race) of exclusion, and the degree to which social influence, authority expectations, and cultural norms explained children's judgments. There were also significant differences depending on the gender, age, and ethnicity of the participants. The findings support our theoretical proposal that exclusion is a multifaceted phenomenon and that different forms of reasoning are brought to bear on the issue. This model was drawn from social-cognitive domain theory, social psychological theories of stereotype knowledge and intergroup relationships, and developmental studies on peer relationships. The results contribute to an understanding of the factors involved in the developmental emergence of judgments about exclusion based on group membership as well as to the phenomena of prejudice, discrimination, and the fair treatment of others. PMID- 12557596 TI - Is it ever ok to exclude on the basis of race or gender?: The role of context, stereotypes, and historical change. PMID- 12557597 TI - [The history of Metasul]. AB - The first generation, metal-on-metal articulation from cobalt-chromium-molybdenum alloy produced by Sulzer Ltd. has been in use for 40 years. Former drawbacks concerning geometry, tolerance or metallurgy have been eliminated in the second generation METASUL components used since 1988. These are also free of disadvantages, such as potential fracture, shown by other hard-on-hard bearing couples. For 14 years now METASUL components have not been responsible for any repeat operation due to loosening. Both clinical and experimental outcomes show that yearly wear has been negligible (in the order of microns) and this allows us to expect the metal-on-metal coupling to function for periods not a achieved so far by any other hip prosthesis. Hip replacements based on METASUL are better than those involving metal-on-polyethylene or ceramic-on-polyethylene. Up to now the need for repeat operations has not been associated with the MATASUL coupling system. Loosening has always been due to other reasons. It can therefore be expected that, in choosing prosthetic devices and surgical techniques, METASUL hip replacements will play a leading role. PMID- 12557598 TI - [A less invasive retropleural-retroperitoneal approach to the thoraco-lumbar spine]. AB - Authors describe the technique of anterior less invasive approach to the treatment of thoracolumbar spine. The advantage of the mentioned approach is the treatment of the anterior spinal column without opening the thoracic cavity. MATERIAL: In the period between 1996-2000 the described method was used in 46 patients. The average age of patients was 49 years (range, 22 to 67 years). There were 30 men and 16 women. Most frequently affected was L1-fractures in 29 patients, Th12 was handled in 16 patients and Th11 only in one case. METHOD: The surgical technique is based on the anterior approach to thoracolumbar spine without opening of the pleural cavity. RESULTS: The evaluation included 46 patients. The surgical time ranged between 100 minutes and 180 minutes. Blood losses ranged between 300 and 2000 ml. Pleural cavity was opened in three cases. Thoracic drainage was not applied in any patient from the followed-up group. Neither preoperative nor postoperative complications were recorded. In all cases allograft was used without complications. DISCUSSION: The described method is one of the possibilities of treating the anterior spinal column in the region of thoracolumbar spine in addition to the classic Hodgson approach or thoracoscopic treatment. CONCLUSION: In the given group of patients the authors' department introduced the treatment of the anterior column of thoracolumbar spine without opening pleural cavity and without the necessity of draining the thoracic cavity. It is a less invasive method as compared to the classic Hodgson approach. The alternative to the treatment of the anterior spinal column in the thoracolumbar region is the thoracoscopic method. PMID- 12557599 TI - [Use of bioceramics in the treatment of fractures of the thoraco-lumbar spine]. AB - PURPOSE OF THE STUDY: The primary reduction and stabilization of all types of injury to the thoracolumbar spine is currently performed from the posterior approach by an internal, transpedicular fixator. The exceptions are type A fractures, according to the AO classification, that can primarily be treated from the anterior approach. The aim of the study was to assess the effect of BAS-O bioceramic granules, inserted by transpedicular approach, on the development of post-operative kyphosis of the segments injured. MATERIAL: Between 1997 and 2001 we treated anterior spinal column fractures, using bioceramic granules in combination with an internal fixator, in 53 patients (40 men, 13 women: age 26 to 69 years; average, 42.3 years) at the Department of Orthopedics and Traumatology of the Third Faculty of Medicine and FNKV in Prague. However, only 42 patients (33 men, 9 women; age 28 to 67 years; average, 41.6 years) who had had the metal fixator removed more than six months previously were included in the study. The fixator was removed in the range of 10 to 24 months after the primary operation. The sample was divided into two groups; one with bioceramic material inserted in the body of the damaged vertebra (20 patients) and the other bioceramic granules implanted in both the vertebral body and the intervertebral space (22 patients). METHODS: In all the patients, the standard procedure included transpedicular application of Schanz's screws and an USS FS fixator (Synthes). If necessary, distraction of the segment injured and the correction of lordosis were carried out. Further transpedicular procedures to treat the anterior spinal column were as follows: If the vertebral body was injured without destruction to the intervertebral disc, a bent elevator was inserted through the vertebral foramen and the intervertebral joints were reduced. At the same time, a cavity for application of bioceramic granules was created. These were inserted, using a funnel and a pusher, in the anterior part of the injured body. If the fracture involved a destroyed intervertebral disc, the disc was removed, the vertebral end plate of the adjacent vertebra was perforated and bioceramic granules were inserted in both the vertebral body and intervertebral space. Subsequently, spongeous bone grafts were harvested from the ilium and massively applied to the previously decorticated transverse and articular processes. RESULTS: In the group of patients who had bioceramic granules inserted in only the vertebral body, the kyphotic angle was on average 10.36 degrees after injury and 2.86 degrees after surgery; therefore, a correction by 13.22 degrees was achieved. At 3 and 6 months after surgery, the kyphotic angle was 2.71 degrees and 2.68 degrees, respectively; at 3 and 6 months after fixator removal, it was 0.67 degree in both instances. In the group of patients with bioceramic granules implanted in both the vertebral body and intervertebral space, the kyphotic angle was on average 9.16 degrees after injury and 4.26 degrees after, surgery; therefore, a correction by 13.43 degrees was achieved. At 3 and 6 months after surgery, the kyphotic angle was -4.11 degrees and -4.00 degrees, respectively; at 3 and 6 months after fixator removal, it was 2.38 degrees and 2.44 degrees, respectively. DISCUSSION: Our results revealed differences between the patients who had bioceramic granules inserted in only the vertebral body and those who had them also in the intervertebral space. At 6 months after surgery, the first group showed the loss of correction per two segments to be 3.53 degrees on the average, whereas the second group had a loss of 6.70 degrees, i.e., twice as high. This may be explained by a more serious damage to the intervertebral disc in the latter group. Only small differences between the groups were found in the kyphotic angle at both 6 months after surgery and 3 to 6 months after fixator removal. This implied that, in both groups, the loss of correction occurred in the period up to 3 months after removal of the fixator. CONCLUSIONS: Bioceramic granules provide material for replacement of osseous tissue in the body of the vertebra as well as conditions necessary for bone restructuring. The loss of correction per segment is lower by about half in patients treated with bioceramic granules than in those who received a spongeous bone grafts. PMID- 12557600 TI - [Use of bioactive glass ceramics in the treatment of tibial plateau fractures]. AB - PURPOSE OF THE STUDY: The surgical treatment of proximal tibia fractures requires a reduction and fixation of the split condyle, elevation of the depressed interfragment of articular surface. The golden standard is the application of autogenous bone grafts for filling in the defect in the cancellous bone of the tibial condyle. Bioactive glass-ceramics material may be successfully used for the filling. Gass-ceramics BAS-O is biocompatible and bioactive and has also suitable mechanical properties and therefore it was used in combination with autogenous bone marrow in order to speed up osteogenesis. MATERIAL: The study covered 12 patients, 4 men and 8 women, with a fracture of the proximal tibia classified according to AO as type B3.1, B3.2 and C3.2. The average follow-up period was 5.5 years (range, 2 to 8 years). METHOD: After the assessment of the type of fracture and revision of soft tissues the depressed interfragment of articular surface was elevated and the defect was completely filled with BAS-O granules in combination with 10 ml of autogenous bone marrow. The broken condyle was fixed by screws or buttress plate. During the first year the process of healing was checked by radiographs at 3-month intervals. All 12 patients were invited for a follow-up examination where the radiograph of the proximal tibia was assessed in two projections and the clinical condition was evaluated using Rasmussen criteria. RESULTS: According to the Rasmussen criteria 12 patients achieved the value of 21-29 points with the average of 26.4 of 30 possible points. The volume of the BAS-O filling was 7.1-44.3 ccm and did not change throughout the whole period of the follow-up. The depression of an interfragment of articular surface prior to operation was 3-17 mm, average 6.3 mm. The residual depression after the operation was 0-4 mm, average 1.7 mm. No symptoms of loosening, extrusion or surrounding of the BAS-O material by fibrous tissue was found. The integration with the surrounding bone occurred during the first 3 months. DISCUSSION: The bioactive coral hydroxyapatite has been successfully used to fill similar defects by a number of authors. Among the benefits of BAS-O granular system is an easy handling and a fast integration with the bone tissue of the recipient due to the development of a hydroxyapatite layer on its surface after the contact with tissue fluid. Autologous bone marrow was added to the system in order to initiate osteogenesis and a faster healing of the defect. The mentioned material is suitable also from the mechanical viewpoint as neither deformation nor resorption was found in the followed-up patients. We reduced the surgery duration and morbidity and improved the patients comfort. CONCLUSIONS: Part of the surgical management of the fracture of the tibial plateau with a depressed segment of subchondral bone is its elevation. For filling in of the traumatic defect in the cancellous bone of the proximal tibia the bioactive, glass-ceramics, granular material BAS-O may be successfully used. Mechanical and chemical properties (compression strength, non-resorption) ensure its stable position in the filling as well as a constant volume. Osteointegration was accelerated by adding 10 ml of autologous bone marrow. The use of the glass ceramics material reduces the surgery duration, improves the patient's comfort and decreases the probability of incidence of complications. PMID- 12557601 TI - [Intra-articular treatment with Hyalgan in osteoarthrosis of the knee joint: experience in clinical practice in the Czech Republic]. AB - PURPOSE OF THE STUDY: To evaluate safety and efficacy of intraarticular (IA) hyaluronic acid (HA; Hyalgan in current medical practice. MATERIAL AND METHODS: Multicenter, prospective, opened, observational study included 601 patients in 35 centers in Czech Republic. Patients had analgesics and NSAIDs on demand as rescue analgesia, but i. a. corticosteroids were not allowed. Patients received 1 series of 5 IA injectious of HA into one knee. Outcome measures were: pain at rest and on walking, patients global and assessment of therapy, Lequesne's index, consumption of NSAIDs. The patients were followed for 17 weeks (12 weeks after the last injection). RESULTS: Patients improved in all outcome measures in week 5 (1 week after the last injection) and this improvement remained constant till week 17 (p < 0.001). The patients judged activity level as better or much better for 78% of treated kness in week 5 and 85.7% in week 17. The mean consumption of rescue NSAIDs dropped fom 4 days/1 week before treatment to 1 day/week during and after Hyalgan therapy. There were no severe side effects reported. Injection site pain occurred in 6.8% patients and in 1.45% for injection, and resulted in 1.4% in withdrawal. CONCLUSION: Hyalgan provided good clinical benefits and acceptable safety profile in current medical practise. The consumption of NSAIDs dropped considerably and therefore contributed to favourable cost/benefit ratio of Hyalgan therapy. PMID- 12557602 TI - [DHS (Dynamic Hip Screw) osteosynthesis in an atypical situation]. AB - The author describes a spiral fracture of the distal femur with a fragmented zone that occurred in the place of the distal end of a splint applied because of the previous treatment of a subtrochanteric fractures by osteosynthesis. Atypical approach including a DHS implant was used to treat the spiral fracture. By literature search it was found that this therapy had not previously been described. Different options for stabilization of this fracture are discussed and it is concluded that, for this indication, the use of the DHS system was the most convenient. The author sets rules for the application of a DHS implant as follows: 1) the procedure should be very fast and carried out from a small approach; 2) the fracture should be spiral or oblique and should permit compression of fragments by means of a sliding screw; 3) another non-removable implant should be present in the femur. He concludes that, if not all three criteria are met, any of the conventional methods of osteosynthesis should be used. PMID- 12557603 TI - [Occult bone trauma detected by magnetic resonance imaging]. AB - Authors deal with a case of a 23-year old women presented with a sport-related injury of the left knee joint. Initially seen in the surgical department, patient presented with pain, swelling, and decreased range of motion of the left knee. Conventional radiographs of the left knee showed no evidence of fracture or osseous abnormality, while the puncture of the joint proved haemarthros. One day later surgical examination demonstrated repeatedly pain and swelling of the left knee expanding to the proximal third of the leg. Repeated puncture exhibited haemorrhagic synovial fluid containing fat tissue, lipohaermarthos. With regard to the contradiction between the surgical examination and negative radiographs, the MRI examination was recommended. MR images revealed linear area of abnormal signal intensity in the medial condyle of the tibia indicating subchondral fracture of the posterior margin of the medial tibial condyle. This finding was associated with increased volume of the haemorrhagic intraarticular fluid. The menisci, intraarticular and capsular ligamentous apparatus was noted to be intact. The control MR examination performed six weeks later demonstrated healed fracture with surrounding bone marrow edema. The importance of non-invasive MR examination for revealing of the occult bone fractures is stressed. PMID- 12557604 TI - [European Hip Society 2002]. PMID- 12557605 TI - More on IDET. PMID- 12557607 TI - Medical costs--what's driving them up? PMID- 12557606 TI - Dilemmas in geriatrics: scenario 6 results. PMID- 12557608 TI - Economic crisis of breast imaging. PMID- 12557609 TI - A genetic approach to the problem of chronic wasting disease in Wisconsin. PMID- 12557610 TI - The effectiveness of a follow-up program at improving HIV testing in a pediatric emergency department. AB - BACKGROUND: HIV infection is on the rise in the adolescent population, with the most common risk for infection being sexual exposure for youth aged 15-24 years. Therefore screening for HIV infection should be performed on adolescent patients presenting to the emergency department (ED) with signs or symptoms of sexually transmitted disease (STD). Lack of opportunity for follow-up of HIV test results may prevent ED physicians from obtaining this test. OBJECTIVE: Our objective was to compare the rate of HIV testing in patients evaluated in the ED for STDs before and after the implementation of a follow-up program. METHODS: We conducted a retrospective chart review of all patients 12-18 years of age seen in the ED between April 1, 1993 and March 31, 1997 with ICD-9 codes for STD. The follow-up program started April 1, 1995. Rates of HIV testing were compared between the two years before and after the institution of the follow-up program to determine whether the follow-up program had an impact on the rate of HIV testing. RESULTS: We reviewed 862 records. Before the follow-up program was in place, 86 (18%) of the patients tested for STD in the ED were also tested for HIV and only 7 (8.1%) were called back with their results. After the follow-up program was in place, 100 (27%) of the patients tested for STD in the ED were also tested for HIV. Attempts were made to notify 57 (57%) of these patients of their HIV test results. Coincident with the implementation of the follow-up program, both the rate of HIV testing and patient notification of HIV results increased (p < 0.05). CONCLUSIONS: Coincident with the institution of an appropriate follow-up system, HIV testing in the ED increased and follow-up of these patients improved. However, further steps should be taken to improve the HIV testing in the pediatric ED. PMID- 12557611 TI - Chiari type I malformation: overview of diagnosis and treatment. AB - Chiari Type I malformation (Chiari I) is a congenital hindbrain anomaly characterized by downward displacement of the cerebellar tonsils through the foramen magnum. This can lead to compression of cerebellar components, the lower brainstem, and the upper cervical spinal cord. In turn, a variety of neurological deficits and permanent nervous system damage may evolve. This review article discusses the etiology, diagnosis, and treatment of patients with Chiari I malformation. Its protean manifestations cause significant overlap with multiple sclerosis, chronic fatigue syndrome, and numerous other conditions. Accordingly, the diagnosis of Chiari I is difficult to establish by clinical evaluation alone. Demonstration of the characteristic hindbrain abnormalities, however, is easily accomplished with magnetic resonance imaging. Neuroimaging should therefore be considered in patients with cerebellar, brainstem, and cervical cord dysfunction. Surgical treatment is indicated in symptomatic patients with radiographic evidence of hindbrain abnormalities. Posterior fossa decompression has also been performed in patients with fibromyalgia and chronic fatigue syndrome based solely on overlapping symptoms with Chiari I. This practice remains controversial. Appraisal of surgical outcome requires postoperative neuroimaging and long-term patient follow-up to assess the permanency of improvement. Preliminary study results of the impact of surgical technique on patient outcome are reported. Ongoing research is devoted to a better understanding of the pathophysiology of Chiari I malformation and the development of more effective medical and surgical treatments. PMID- 12557612 TI - Risperidone induced galactorrhea: a case analysis. AB - We report on the occurrence of galactorrhea and amenorrhea associated with prolactin elevation after 6 months of treatment with risperidone. The symptoms dramatically resolved with discontinuation of the drug. We discuss the possible mechanisms and clinical sequelae of elevated prolactin levels. PMID- 12557615 TI - Current therapy of the right ventricle myocardial infarction. AB - BACKGROUND: Acute myocardial infarction of the right ventricle (AMI-RV) is a separate subgroup within the scope of inferoposterior infarction of the left ventricle. It still represents the population of patients at high risk due to numerous, often hardly predictable complications and high mortality rate. METHODS: In fifteen-year period (1987-2001) 3,765 patients with the acute myocardial infarction (AMI) of different localizations of both sexes--2,283 males and 1,482 females of the average age 61.4 +/- 4.6 years were treated in our institution. Anterior myocardial infarction was diagnosed in 2,146 (56.9%) patients, inferior in 1,619 (43.1%) patients, out of whom right ventricular infarction (RVI) was confirmed in 384 (23.7%). Thrombolytic therapy was administered in 163 (42.4%) patients with RVI, and in 53 (41.7%) of these patients balloon dilatation was performed with coronary stent implantation in 24 (45.2%). RESULTS: Favorable clinical effect of the combined thrombolytic therapy and percutaneous transluminal coronary angioplasty (PTCA) was achieved in 51 (96.1%), and in only 2 (3.9%) of patients the expected effect wasn't achieved. Myocardial revascularization was accomplished in 6 (3.6%) and 1 patient died. In 3 (3.4%) patients primary balloon dilatation with the implantation of intracoronary stent was performed within 6 hours from the onset of anginal pain. In the other group of 221 (57.5%) patients with RVI who did not receive thrombolytic therapy, or it had no effect, 26 (11.7%) patients died, which indicated the validity and the efficacy of this treatment (p < 0.01). In the whole group of patients with myocardial infarction of the right ventricle 31 (8.1%) died; in the group that received thrombolytic therapy and PTCA 5 (3.1%) died, while in the group treated in a conservative way 26 (11.7%) died. CONCLUSION: Combined therapy was successful in the treatment of patients with RVI and should be administered whenever possible, since it was the best prevention of life-threatening complications and the decrease in the mortality of those patients. PMID- 12557614 TI - Production of TNF-alpha by skin explants of dinitrochlorobenzene-challenged ears in rats: a model for the evaluation of contact hypersensitivity. AB - BACKGROUND: Contact hypersensitivity (CHS) is a local inflammatory response of the skin following challenge of hapten-sensitized animals. It is the consequence of cell infiltration of derm and the release of inflammation mediators, among which Tumor necrosis factor-alpha (TNF-alpha) is one of the most important factors. The intensity of the inflammation could be quantified by ear swelling which is the classical manifestation of the reaction. This study was testing the working hypothesis that levels of TNF-alpha in skin organ culture medium should correlate with the intensity of CHS reaction measured in vivo by ear swelling assay, and with the density of dermal infiltrate in ear skin samples. In order to test the working hypothesis, the intensity of inflammatory reaction following challenge was evaluated by classical measurements of ear swelling, by the determination of TNF-alpha levels in culture fluids of ear skin following epicutaneous application of dinitrochlorobenzene (DNCB) into the ears of sensitized animals. METHODS: Animal model of CHS reaction to DNCB in Albino Oxford rats was used as described. Ear swelling was quantified in percentage terms as the difference in thickness between the challenged and nontreated ears of the same animal. Dermal infiltrate density in histopathologically analyzed samples of ear skin was evaluated by computer-assisted image analysis. Ear skin samples were cultured in standard medium for 24 h, and TNF-alpha concentration in the conditioned medium was subsequently determined with ELISA test. RESULTS: Dose dependent increase in the density of the dermal infiltrate and in TNF-alpha in CM were noted following the application of 0.65%, 1.3% and 2.6% of DNCB to the ears of previously sensitized rats. The correlation between ear swelling and the levels of TNF-alpha (r = 0.933, p < 0.001) in CM, and between ear swelling and dermal infiltrate density (r = 0.916, p < 0.001) was found. Correlation was also found between the density of the dermal infiltrate and the levels of TNF-alpha (r = 0.865, p < 0.001). CONCLUSION: Presented data suggested that skin-organ culture system and the quantification of inflammatory mediators might be used for the evaluation of contact hypersensitivity reaction and its intensity. PMID- 12557616 TI - Age and body mass related changes of cardiovascular risk factors in women with polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is considered a metabolic disorder closely related to obesity, insulin resistance (IR), hyperinsulinemia and unfavorable lipid profile, all increasing the risk for the occurrence of cardiovascular diseases. The aim of this study was to assess age and body mass index (BMI) related changes of cardiovascular risk factors in 90 women with PCOS. The cut-off age point was 30 years and for BMI 27.8 kg/m2. In all patients systolic and diastolic blood pressure (BP), metabolic parameters comprising values of glucose and insulin during oral glucose tolerance test (OGTT), and basal lipid values were determined. Significant increase in blood pressure (BP) indices, basal insulin values and insulin resistance (IR) assessed by HOMA model were observed with aging and the increase of BMI, while the parameters of glucose metabolism, total cholesterol and triglycerides were significantly elevated only with aging. However, the correlation between the indices of arterial blood pressure, and lipid and glucose metabolism parameters occurred only in patients over 30 years of age, pointing to the causative relation and the consequent deterioration of IR and lipid profile with aging, influencing cardiovascular function in women with PCOS. PMID- 12557617 TI - [Reconstruction of the thumb using a forearm osseofasciocutaneous reverse flap]. AB - This paper presents the experiences of the thumb reconstruction with osteofasciocutaneous reverse flap (OFCR flap). In the period between 1987 and 2000 the OFCR flap was used in 15 patients. The youngest of them was 18 and the oldest was 38 years of age. The average age was 25.4. All the patients had posttraumatic amputations. Defects on proximal phalangae and a part of metacarpal bone occurred in two cases. In one case there was an amputation on the base level of proximal phalanx and the metacarpophalangeal (MPH) joint was preserved. In all cases of reconstruction the OFCR flap was used, which included antebrachial skin nervs that were anastomosed with digital nerv. The flap nutrition was carried out through the reverse circulation of a. radialis, and the venous drainage through the comitant vein of a. radialis. Superficial veins were not anastomosed. Secondary defects were covered with a free skin graft. All the flaps survived. The bone graft was healed in the period of eight weeks. The sensibility of this flap was regained in the period of three to six months after the surgery. The distance of two-point discrimination (TPD) was increased for 30% compared to the same region on the other hand after six months. The opposition of the reconstructed thumb to the other fingers was possible, as well as abduction, adduction and normal grasp. The method of reconstruction of the amputated thumb with the OFCR flap was better than other classical methods because it allowed the reconstruction of all the structures in one surgical operation. The sensibility that was regained represented good protection from injuries. There were no functional damages on the secondary defect. The esthetic result was not good due to the lack of a fingernail. PMID- 12557618 TI - [Co-induction in ambulatory anesthesia]. AB - BACKGROUND: Co-induction in anesthesia is very useful: synergistic effects of two inductional drugs may lower the dose regimen and the incidence of adverse effects. The aim of this study was to investigate and compare two anesthesiological techniques for short-lasting gynecological procedures in outpatient anesthesia. A total of 80 patients scheduled for surgical termination of pregnancy were randomly assigned into two equal groups--control and co induction group. METOHDS: The first group of patients received atropine 0.5 mg i.v., alfentanil 0.5 mg i.v. and propofol as a fractionated i.v. bolus until the loss of eyelash reflex. The second group received atropin 0.5 mg, alfentanil 0.5 mg, midazolam 3 mg and propofol in the same manner as the first group. Anesthesia was maintained with propofol increments. Cardiovascular parameters, parameters of post anesthesia recovery and the adverse effects were registered. RESULTS: In patients receiving midazolam inductional dose of propofol was significantly lower, whereas cardiovascular parameters were not significantly different. The recovery after anesthesia was slightly longer after co-induction, but it was not of great clinical significance. The reduction of the adverse effects was found in the co-induction group. CONCLUSION: The results of the study showed that co induction of midazolam-propofol in comparison with propofol alone for outpatient anesthesia had the following advantages: the reduction of propofol dose, better quality of anesthesia and the reduction of the adverse effects. Recovery was faster in the group that didn't receive midazolam, but it was not of great clinical significance. The conclusion is that co-induction with the combination midazolam-propofol has the advantage in outpatient procedures. PMID- 12557619 TI - [Diagnosis of the Mikulicz-Sjogren syndrome using biopsy of the minor salivary glands]. AB - BACKGROUND: Sjogren's syndrome (SS) represents autoimmune disease characterized by chronic inflammation, destruction and insufficiency of exocrine glands, particularly salivary and lacrimal glands, accompanied by dryness of mouths and eyes. Diagnostic work-up involves clinical laboratory tests, radiography, scintigraphy and bioptic histopathological examination. Examination of small salivary glands in the biopsy of the lower lip represents a "golden standard" of diagnosis of SS, concerning the fact that the growth and the disfunction of salivary and lacrimal glands occurs in different pathologic states. METHODS: Resected specimens of the lower lip were obtained from 47 patients with clinical diagnosis of SS. After standard histopathological treatment, slices were hematoxylin and eosin stained. Immunohistochemistry against smooth muscle actin was performed using LSAB+ method (AHSMA-M7558, DAKO 1:50). On the basis of generally accepted histopathological diagnostic criteria the results were categorized as: findings suspicious for SS; findings compatible with the diagnosis of SS (mild, moderate and high degree of inflammation); nonspecific inflammatory reaction and nonrepresentative biopsy samples. RESULTS: Diagnosis of SS was confirmed in 32% of cases. In 2% of cases findings were suspected for SS, in 36% of cases findings were compatible with the diagnosis of nonspecific inflammation, and in 30% of cases material was not representative. CONCLUSIONS: By the biopsy of salivary glands of the lower lip the diagnosis of SS was confirmed in 50-60% of cases. Upon the precise diagnostic criteria it was also possible to determine the intensity of inflammation and tissue destruction in SS and identify other pathological conditions, which justified the biopsy. Surgical technique had to be adequate in order to obtain representative number of small salivary glands. In the presented material 30% of specimens were nonrepresentative which was very high percentage compared with literature data. This was most probably the reason why the diagnosis of SS was confirmed in only 32% of cases, i.e., in every third patient. PMID- 12557620 TI - [Manufacture of tetracaine hydrochloride tablets using direct compression and moist granulation]. AB - The results of quality testing of lozenges made by direct compression and by wet granulation procedure (Ph. Jug. IV) are presented. The aim was to determine hardness, friability and disintegration of the lozenges. Considering the fact that the produced tetracaine hydrochloride lozenges were not transported, but were made according to the needs of the Clinic of Gastroenterology, Military Medical Academy, time of disintegration, not hardness and friability, was the priority in making a choice of formulation. That is why tetracaine hydrochloride lozenges made with 3.5% solution of carmellose sodium, which have the longest disintegration were chosen, which made possible the longest contact of the local anesthetic with mucous membrane of mouth and throat. PMID- 12557621 TI - [Principles of care of thoracic war injuries]. PMID- 12557622 TI - [The blood-brain barrier]. PMID- 12557623 TI - [Clinical and microbiologic aspects of blood cultures]. PMID- 12557624 TI - [ICurrent knowledge about molecular mechanisms of regulation of iron homeostasis]. PMID- 12557625 TI - [The inflammatory reaction and adhesion molecules in cerebral ischemia]. PMID- 12557626 TI - Non-Hodgkin lymphomas of oral cavity. AB - Non-Hodgkin lymphomas (NHL) often show up in an extranodal pattern, especially in the head and neck. Intraoral locations are much less frequent, particularly when they are single. This, in turn, can lead to a prolonged diagnosis and even to inadequate treatment. Different patients with initial extranodal location of NHL which were not previously diagnosed and in which it was manifested only intraorally are presented in this paper. These cases are presented together with the additional examinations used for the early diagnosis and with the corresponding clinical pictures, as well as with the overview of other cases from the available literature. PMID- 12557628 TI - [History of Serbian and Yugoslav military psychiatry: founding and development of the Clinic of Military Psychiatry from 1932 to 2002]. PMID- 12557627 TI - [Treatment of acute myocardial infarct with ST segment elevation with a combination of fibrinolytic therapy and abciximab]. AB - BACKGROUND: According to current knowledge, the best way to treat the acute myocardial infarction with ST elevation is primary transluminal coronary angioplasty, which can be performed only in the best equipped tertiary cardiology centers. As it was known that atherothrombosis wais the essence of the acute coronary syndrome we wanted to examine the efficacy and safety of combined therapy of tissue plasminogen activator and glycoprotein IIbIIIa platelet receptor antagonist abciximab. METHODS: The case is reported of combined abciximab and accelerate schedule of t-PA reperfusion therapy in a young patient with the extensive anterior acute myocardial infarction. Activated partial thromboplastin time and platelet count were regularly measured during therapy. RESULTS: The combination of these two drugs did not cause any complication in our patient. According to early noninvasive parameters, successful reperfusion was achieved. Postinfarction period was without complications. Coronary angiography was performed 15 days after and was without pathological findings. Eighteen months later the event patient had neither chest pain, nor other complaints with slightly reduced R waves in middle precordial leads and hypokinesis of anterior apical segment of the left ventricle showing the signs of important systolic function impairment. CONCLUSION: Controlled studies are needed to prove the safety and the benefit of such combined reperfusion therapy and to show which kind of treatment is appropriate in every case considering the patient conditions and the facilities of coronary care unit. PMID- 12557629 TI - [History of the Academy of Military Medicine: IV--Internal medicine]. PMID- 12557630 TI - Violent behavior among urban youth attending alternative schools. AB - This study described violent behavior and aggression among youth attending alternative schools, and examined sociodemographic factors associated with such violence. The study involved 494 students attending 10 alternative schools in Houston, Texas. Data were collected between November 2000 and February 2001 by audio-enabled laptop computers equipped with headphones. Students self-reported an average of 11.8 aggressive acts during the week prior to the survey. Students reported a 30-day weapon carrying prevalence of 22.7%; 30-day gun carrying prevalence of 11.1%; 30-day knife or club prevalence of 17.2%; 12-month fighting prevalence of 50.6%; and 12-month prevalence of injuries due to fighting of 6.5%. Association between demographic variables, self-reported aggressive behavior, and other forms of aggression was examined using multivariate logistic regression. Students were divided into four mutually exclusive violence-related categories: no fighting and no weapon (referent), fighting only, carrying weapon only, fighting and weapon carrying. Student aggression was significantly associated with fighting and weapon carrying, with incremental increases at each level (OR 1.1 per unit of increase, 95% CI 1.1-1.2) and in presence of the covariates. Among Houston's alternative school students, the prevalence of violent behavior (weapon carrying, gun carrying, knife or club carrying, fighting, and treatment by a doctor or nurse for injuries) is higher compared to regular high school students. Aggression related strongly to weapon carrying and fighting in the sample. Data indicate alternative school youth urgently need prevention and treatment programs to help them live in safer environments. PMID- 12557631 TI - Residential adolescent substance abuse treatment: recommendations for collaboration between school health and substance abuse treatment personnel. AB - Thousands of youth participate in residential substance abuse treatment each year. After completing treatment, many of these youth return to school. This study analyzed the process of substance abuse treatment at the Generations Youth Program, and identified opportunities for collaboration with school health personnel. A qualitative case study design was employed. Analysis of the treatment process revealed the primary goal of treatment was to assist youth in developing a belief in their ability to remain sober. This goal was achieved through a peer support network, development of self-control, and acquisition of treatment knowledge. Youth who develop these skills are successfully discharged from treatment and return to home and school. Opportunities exist for collaboration between school health and substance abuse treatment personnel to enable discharged youth to remain drug free. PMID- 12557632 TI - Self-reported school difficulties and tobacco use among fourth- to seventh-grade students. AB - This study examined the relationship between academic and behavioral difficulties at school, and tobacco use in students. Participants included 1,219 students in fourth to seventh grade at the time of enrollment. Interviews were repeated eight months later with 85% of baseline participants. Telephone interviews assessed use of cigarettes, cigars, and chewing tobacco; students also were asked if they liked school, how often they got in trouble at school, and how well they were doing in school. At baseline, students reporting school difficulties were 1.4-5.6 times more likely to report a lifetime history of cigarette, cigar, and chewing tobacco use relative to students who did not report these difficulties. Average to below-average academic performance at baseline was predictive of new cigarette use at the eight-month follow-up (Relative Risk = 3.35; 95% Confidence Interval = [1.36, 8.22]). Self-reported school difficulties are associated with lifetime use of all major forms of tobacco and are predictive of future cigarette use in fourth- to seventh-grade students. PMID- 12557633 TI - Do perceived cues, benefits, and barriers to physical activity differ between male and female adolescents? AB - A four-page survey was administered to 535 adolescents at two single-sex (one male, one female) high schools in Cincinnati, Ohio, to examine whether perceptions of physical activity differed by gender. More specifically, the survey assessed perceived cues, benefits, and barriers to exercising. Results indicated that the most helpful cue to physical activity for both female and male students was "having a friend to exercise with." The most commonly reported benefit of exercising among females was "to stay in shape," whereas the most commonly reported benefit to exercising among males was "to become strong." Among females, the most common barrier to exercising was "having no time to exercise," whereas males were most likely to report "wanting to do other things with my time." Multivariate analyses of covariance revealed that perceived cues, benefits, and barriers to physical activity differed significantly based on gender. Recommendations on specific strategies to increasing male and female adolescent physical activity levels are offered. PMID- 12557634 TI - A sharper image for school health education: Hawaii's "seven by seven" curriculum focus. PMID- 12557635 TI - Emotional moments. PMID- 12557636 TI - A comprehensive approach to improve the oral health of students in Edmonton, Alberta, Canada. PMID- 12557637 TI - Health sector reforms: implications for sexual and reproductive health services. PMID- 12557638 TI - Understanding health sector reforms and sexual and reproductive health services: a preliminary framework. AB - To understand the implications of health sector reforms for sexual and reproductive health services, there are three major dimensions to consider. The first two relate to the context in which health sector reforms are introduced: the characteristics of the health system, and that of reproductive and sexual health services located within it. The third dimension has to do with the content and scope of health sector reforms introduced into this context, and the actors and processes through which it is introduced. The content, scope, actors and processes have in turn to be located and understood within the larger geopolitical context and the position within it of the country under consideration. PMID- 12557639 TI - An overview of changing agendas in health sector reforms. AB - This paper discusses health sector reforms and what they have meant for sexual and reproductive health advocacy in low-income countries. Beginning in the late 1980s, it outlines the main macro-economic shifts and policy trends which affect countries dependent on external aid and the main health sector reforms taking place. It then considers the implications of successive macro-economic and reform agendas for reproductive and sexual health advocacy. International debate today is focused on the conditions necessary for socio-economic development and the role of governments in these, and how to improve the performance of health sector bureaucracies and delivery systems. A critical challenge is how to re-negotiate the policy and financial space for sexual and reproductive health services within national health systems and at international level. Advocacy for sexual and reproductive health has to tread the line between a vision of reproductive health for all and action on priority conditions, which means articulating an informed view on needs and priorities. In pressing for greater funding for reproductive health, advocates need to find an appropriate balance between concern with health systems strengthening and service delivery and programmes, and create alliances with progressive health sector reformers. PMID- 12557640 TI - Between intent and achievement in sector-wide approaches: staking a claim for reproductive health. AB - Since 1995, sector-wide approaches (SWAps) to health development have significantly influenced health aid to developing countries. SWAps offer guidelines for new partnerships with international donors led by government, new relationships between donors and shared financing, development and implementation of agreed packages of health sector reforms. These structural and funding changes have significant implications for reproductive health. The early experience of SWAps suggests that the extent of donor commitment is constrained for administrative, philosophical and political reasons, with vertical programmes (including those relevant to reproductive health) protecting their 'core' business, and reproductive health, as an integrative concept, lacking strong advocates. Defining the sector in terms of government health systems focuses resources on building effective district health systems, but with uncertain outcomes for elements of reproductive health that depend on multi-sectoral strategies, e.g. safe motherhood. The context of the reforms remains a determining factor in their success, but despite savings available through increased efficiencies and coordinated services, the total per capita expenditure on health to ensure minimum clinical and public health services often remains beyond the budget available to least developed nations. Despite this, many of the elements of SWAps--government leadership, new donor relationships, better coordination, sectoral reform and service integration--offer the potential for more effective and efficient health services, including those for reproductive health. PMID- 12557641 TI - Objectivity in priority setting tools in reproductive health: context and the DALY. AB - The Disability Adjusted Life-Year (DALY) heralded a new age in the development of an evidence base for priority setting and resource allocation. The DALY was intended to represent sound measurement of the incidence and prevalence of conditions in a process by which burden of disease would be determined, health conditions prioritised and interventions evaluated. However, in the reification of objectivity, sight was lost of critical aspects of health and disease, namely that they are not independent of the context in which they occur. This is powerfully illustrated by many reproductive health conditions. Priority setting and resource allocation exercises need to take into account both objective measures and contextually relevant factors. Based on comparative data that highlights the effect of the development gradient on the burden of disease, this paper discusses the implications of context in the assessment of population health and priority setting, with a focus on reproductive health. PMID- 12557642 TI - The politics of priority setting for reproductive health: breast and cervical cancer in Ghana. AB - Priority setting for reproductive health is affected by health sector reform policies, the often politically charged nature of issues such as abortion, decreasing funding for reproductive health activities and the broad agenda of the ICPD Programme of Action. This paper examines the influence of political and organizational factors on national priority setting for reproductive health and argues that existing priority setting tools such as disability-adjusted life years and cost-effectiveness analysis do not consider the influence of politics on the priority setting process or account for the interpretation of evidence in priority setting. It suggests that priority setting tools can be strengthened by incorporating empirical measures of political and organizational attention to an issue, and through a new measure--policy priority. The paper applied this new measure to a case study of attention to breast and cervical cancer in Ghana from 1990-97, illustrating how traditional priority setting methods cannot explain the priority given to breast cancer in Ghana. It demonstrates how local politics can trump scientific and economic evidence and suggests that the priority setting process can have unforeseen equity and social implications. It concludes by arguing that the policy priority measure provides a more complete picture of reproductive health priorities and is useful for better understanding the implications of the priority setting process for reproductive health. PMID- 12557643 TI - Universal coverage and its impact on reproductive health services in Thailand. AB - Thailand has recently introduced universal health care coverage for 45 million of its people, financed by general tax revenue. A capitation contract model was adopted to purchase ambulatory and hospital care, and preventive care and promotion, including reproductive health services, from public and private service providers. This paper describes the health financing system prior to universal coverage, and the extent to which Thailand has achieved reproductive health objectives prior to this reform. It then analyses the potential impact of universal coverage on reproductive health services. Whether there are positive or negative effects on reproductive health services will depend on the interaction between three key aspects: awareness of entitlement on the part of intended beneficiaries of services, the response of health care providers to capitation, and the capacity of purchasers to monitor and enforce contracts. In rural areas, the district public health system is the sole service provider and the contractual relationship requires trust and positive engagement with purchasers. We recommend an evidence-based approach to fine-tune the reproductive health services benefits package under universal coverage, as well as improved institutional capacity for purchasers and the active participation of civil society and other partners to empower beneficiaries. PMID- 12557644 TI - Quality of hysterectomy care in rural Gujarat: the role of community-based health insurance. AB - Community-based health insurance (CBHI) may be a mechanism for improving the quality of health care available to people outside the formal sector in developing countries. The purpose of this paper is to identify problems associated with the quality of hysterectomy care accessed by members of SEWA (Self-Employed Women's Association), an Indian CBHI scheme, and discuss mechanisms that would optimize quality of care. Data on hysterectomy care were collected through a review of 63 insurance claims and semi-structured interviews with 12 providers. Quality of hysterectomy care accessed by SEWA's members varied from potentially dangerous to excellent. Dangerous conditions included operating theatres without separate hand-washing facilities or proper lighting, the absence of qualified nursing staff, performing hysterectomy on demand, removing both ovaries without consulting or notifying the patient, and failing to send the excised organs for histopathology, even when signs were suggestive of disease. Women paid substantial amounts of money, even for poor and potentially dangerous care. In order to improve the quality of care for its members, a CBHI scheme can: (1) gather data on the costs and complications for each provider, and investigate where these are excessive; (2) use incentives to encourage providers to make efficient and equitable resource allocations; (3) contract with providers giving a high standard of care or who agree to certain conditions; and (4) inform and advise doctors and the insured about the costs and benefits of different interventions. In the case of SEWA, it is most feasible to identify a limited number of hospitals providing better quality care and contract directly with them. PMID- 12557645 TI - The role of oversight in the health sector: the example of sexual and reproductive health services in India. AB - This paper examines the role of oversight in influencing the health sector, using examples from sexual and reproductive health services in India. Rather than simply trying to provide services through traditional bureaucratic mechanisms, governments can make use of oversight tools to influence how health care is delivered through the public and private sectors. Three main oversight functions are described: understanding health system performance, deciding when to intervene in the health system and strategizing and implementing change. Governments also need to understand the ethical basis for decisions. The potential for administering oversight through policy-making, disclosing and informing, regulating, collaborating, and strategically subsidising and contracting services in sexual and reproductive health is described. This approach implies an engagement with a broader set of stakeholders in the health sector than is often the case. It requires a set of skills for public officials beyond managing public programmes, and relies on a larger role for other stakeholders and the general public. When applied to reproductive and sexual health, implementation of the full range of oversight functions offers new opportunities to provide more effective, equitable, accountable and affordable services. PMID- 12557646 TI - Chinese maternal health in adjustment: claim for life. AB - Health sector reforms in China, instituted starting in 1985, have centred on cost recovery, with fee-for-service revenue replacing public budget funding. The share of public funding for maternal health services was reduced greatly, forcing an increasing proportion of pregnant women to pay for deliveries and treatment of pregnancy-related complications out of pocket, as most had no health insurance to cover these costs. This study aimed to identify socio-economic variables associated with utilisation of essential maternal health services and linked to health sector reforms in China, with a focus on cost recovery. A retrospective household survey (n = 5756) was carried out in six counties in three provinces of Central China in 1995. Antenatal service utilisation continued to improve in 1990 95, but only in relation to the number of visits, which were pre-paid if the woman was participating in a maternal pre-payment scheme or covered by another health insurance scheme. Significant decreases were found in the utilisation of skilled attendance at delivery and hospital delivery, as well as differences in adverse pregnancy outcomes (miscarriages and stillbirths) between women paying out of pocket and those covered by insurance. This study confirms a strong association between utilisation of delivery services and financing variables of amount of savings in the bank, maternal pre-payment schemes and health insurance. It also shows the critical importance of out-of-pocket, fee-for-service payments for maternity care as a barrier to the utilisation of these services. PMID- 12557647 TI - Privatisation of health services and the reproductive health of rural Chinese women. AB - China's rural health care system has undergone major changes since the early 1980s, when the country began privatising rural health services. Following fiscal devolution, the rural primary health service was transformed into a fee-for service system, dependent on the availability of local resources. This article reports some of the results of a study undertaken in 1994-96 to examine the impact of privatization on financing, provision and use of reproductive health services by women in two rural counties in Yunnan Province, China. The most common self-reported symptoms of reproductive morbidity were abnormal vaginal discharge and vaginal tears during home delivery, which went mostly untreated. Hospital-based delivery and use of antenatal care was very low, adversely affected by costs and perceived low quality. Service quality was affected by low investment in training, maintenance and supervision of workers. Most of the burden for maternal and child health care fell on local health workers, yet resources for these services had declined from 1985 to 1995. Only support for family planning services, which were funded and provided separately, had increased. Rural women's reproductive health needs were inadequately attended to by rural health services following reforms. Our data has helped to increase attention to those needs within planned reform efforts. PMID- 12557648 TI - Women's health, changes and challenges in health policy development in Lithuania. AB - Health is a sensitive mirror of social circumstances. This paper looks at the situation of women's health in Lithuania in the context of the social, political and economic transition in the country following independence in 1990, and reforms to the health system. Data since 1990 show that considerable social and demographic inequalities in the health of women exist in Lithuania, with low educated women and those living in rural areas in the most unfavourable situation, including in relation to reproductive health. Reproductive health issues have received some recognition in recent years, with the main attention and resources directed to the development of a Maternal and Child Health Programme, especially perinatal care and the organisation of neonatology services, which has resulted in a notable decrease in maternal, perinatal and infant mortality. Services for family planning, abortion, infertility, cervical and breast cancer, and violence against women are under-developed. Non governmental organizations are beginning to be formed to advocate for increased resources and services for reproductive health. Improvements in the health status of Lithuanian women can be expected if attention is paid to social determinants of health. PMID- 12557649 TI - Gender dimensions of user fees: implications for women's utilization of health care. AB - This paper looks at the implications of user fees for women's utilization of health care services, based on selected studies in Africa. Lack of access to resources and inequitable decision-making power mean that when poor women face out-of-pocket costs such as user fees when seeking health care, the cost of care may become out of reach. Even though many poor women may be exempt from fees, there is little incentive for providers to apply exemptions, as they too are constrained by restrictive economic and health service conditions. If user fees and other out-of-pocket costs are to be retained in resource-poor settings, there is a need to demonstrate how they can be successfully and equitably implemented. The lack of hard evidence on the impact of user fees on women's health outcomes and reproductive health service utilization reminds us of the urgent need to examine how women cope with health care costs and what trade-offs they make in order to pay for health care. Such studies need to collect gender-disaggregated data in relation to women's health service utilization and in relation to the range of reproductive health services, taking into account not only out-of-pocket fees charged by public health providers but also by private and traditional providers. PMID- 12557650 TI - Short-changing reproductive health. AB - Health sector reformers, particularly the economists among them, are prone to reject calls for more financial support for reproductive health as "special pleading" even when the argument is made that reproductive health is a basic human right. Reproductive health is not alone in this. Up to now, however, the idea that the right to reproductive health care should take precedence over other health rights has not been put on the table. Economists ask how much interventions cost and whether there is enough money to cover all of the interventions being proposed; if not, they ask which interventions will be funded. Basing decisions about spending for health on evidence about the burden of disease is a way of using the principles of economics to improve health systems performance. However, this methodology poses some special problems for reproductive health as pregnancy is not a disease. Further, economic principles are not the only valid criteria for decisions about health care funding. Those who are concerned about reproductive health need to remain vigilant about the impact of these changes and counter with their own evidence on how they may or may not be contributing to improved health system performance and to reproductive health and rights. PMID- 12557651 TI - ICPD goals and targets worth revisiting in the context of health sector reforms. PMID- 12557652 TI - UN special session on children: Bush Administration continues its attacks on sexual and reproductive health. PMID- 12557653 TI - Counsellors' perspectives on antenatal HIV testing and infant feeding dilemmas facing women with HIV in northern Tanzania. AB - This study investigated the infant feeding advice that counsellors were giving HIV-infected pregnant women in Moshi, Tanzania, the factors they thought had an impact on women's infant feeding choices and their role in influencing these decisions. The data are drawn from in-depth interviews with 16 nurses working as counsellors in their spare time in an antenatal trial of prevention of mother-to child transmission, five local HIV/AIDS counsellors and two medical doctors, whose counselling experience ranged from less than six months to nine years. Informed choice of infant feeding method by HIV-infected women, as recommended by UNAIDS/WHO/UNICEF Guidelines, was seriously compromised by the actual advice given, directive counselling, lack of time to cope with a positive HIV test result, and lack of follow-up support, regardless of socio-economic status. Infant feeding options were not always accurately explained, but counsellors believed most women had little choice but to breastfeed and were unlikely to exclusively breastfeed, despite advice. It was apparent that the risks and benefits of the options open to HIV-infected women were complicated for the counsellors, not only the women. Counsellors needed additional training in non directive counselling and infant feeding options to ensure a better quality of advice-giving and support to follow-up women at home. PMID- 12557654 TI - Mesoglia & microglia--a historical review of the concept of mononuclear phagocytes within the central nervous system. AB - More than a century and a half has elapsed since the first accounts of mesodermal phagocytic elements were proposed within the central nervous system. Over the intervening decades, body and substance were added to this concept through the advancement of histological techniques at the disposal of the researcher and the acute and keen-minded skills of the pathologist. Notable among these pioneering efforts were the contributions of W. Ford Robertson, Santiago Ramon y Cajal, Pio del Rio-Hortega and Wilder Penfield amongst an entire cavalcade of other noteworthy figures. The term 'mesoglia' and 'third element of the nervous system' was bestowed upon these cells towards the beginning of the twentieth century to account for their separate origins from neurons and macroglia. It was later amended by del Rio-Hortega in 1919, to 'microglia' in order to further discriminate between true mesodermal elements and oligodendrocytes, previously regarded as a component of 'mesoglia'. This particular contention sparked much controversy among del Rio-Hortega's peers and resulted in an escalation of fruitful research throughout Europe that eventually declined up to the outbreak of the Second World War. The post-war years were a period of the 'dark ages' that cast doubt on the very existence and nature of microglia, until the 'renaissance' of research was once again rejuvenated in the 1960s, by a new cohort of intrigued minds: Cammermeyer, Blinzinger, Kreutzberg and others who saw in the 'third element' the potential that is now commonly ascribed to microglia: the intrinsic immune effector cells of the CNS. It is now universally accepted that microglia are involved as the first line of rapid defence in any pathology of the nervous system, and as such, present a diagnostic tool for the neuropathologist. Although our knowledge of microglia stems from an extensive body of work conducted over the last two decades, much of the earlier work (pre-1960s) has remained somewhat obscure. This is partly accountable due to the limited availability of translated works, and additionally to the lack of a compendium of these articles. This paper will present a comprehensive overview of the pioneering research on mononuclear phagocytes within the central nervous system, which has direct bearing on our present-day understanding of the concept of microglia. PMID- 12557655 TI - Lesions as therapy: surgical intervention in Parkinson's disease prior to L-dopa. AB - The study of Parkinson's disease has undergone vast changes across its almost 200 year history. Over this period, research scientists have added dramatic detail to their understanding both of the motor system in general and the etiology of Parkinson's disease specifically. This expanded understanding has been facilitated, particularly, by the work of clinicians with the goal of improving treatments designed to ameliorate its symptoms. This article examines the evolution of one particular clinical approach, the production of lesions to segments of the basal ganglia, from its inception, through its "golden era," disuse, and rebirth. PMID- 12557656 TI - Yawning? AB - Since antiquity yawning has attracted a moderate interest among philosophers, psychologists, physiologists, as well as educators, moralists and physicians. Organisms from birds to men and from the womb to the deathbed were found to be displaying it. While sometimes satisfying to the producer, its display is offensive to the lay observer. Hippocrates had it on his lists of useful 'natures.' Aristotle dropped a few words on the matter. Boerhaave elevated its function to the intellect of animals. Haller has commented on its relation to the acoustic system, blood-flow, and baby sleep. Darwin mentioned it in connection with emotional behavior. Some modern authors praised its beneficial effects on respiration and smell. In the 1960s, Ashley Montagu tried to correct the contemporary failure to explain the behavior by the fact of raised CO2 and arterial compression. It also interested some neurologists, especially in its association with the encephalitis lethargica in the 1920s, with 'spasmodic yawning,' with epilepsy, not to speak of hysteria. As to boredom or its stimulus, a 40-page dissertation survives from the court of Frederick the Great of the 18th century condemning idleness, a subject that also inspired Blaise Pascal and William James. But in the Hindu world, public yawning was a religious offense. PMID- 12557657 TI - An American neurologist, pioneer of cerebral localization and treatment of aphasia. PMID- 12557658 TI - The neurognostic question 16 and its answer. PMID- 12557659 TI - [Leaf photosynthetic acclimation of Echinochloa crusgalli grown in rice field to free-air CO2 enrichment (FACE)]. AB - A comparative study was made between the leaves of Echinochloa crusgalli grown at 580 (FACE) and 380 (control, ambient air) mumol CO2 mol-1 air by observation of net photosynthetic rates (Pn) and the contents of soluble protein and the key enzyme of photosynthetic carbon assimilation, Rubisco. When measured at the ambient air CO2 concentration (380 mumol.mol-1), Pn were significantly lower in leaves grown under FACE conditions than that in those leaves grown in the ambient air. This indicates that photosynthetic acclimation to high CO2 occurs in the leaves grown under FACE conditions. Also, the stomatal conductance (Gs) and intercellular CO2 concentration (Ci) were decreased significantly in the leaves grown under FACE. The content of total soluble protein was much lower in the leaves grown under FACE conditions than that the in the control, and the content of Rubisco was also decreased in the FACE leaves, but the difference was not significant. From these results it is deduced that the photosynthetic acclimation to high CO2 in Echinochloa crusgalli leaves grown under FACE conditions is mainly related to the partial closure of stomata and the decrease in soluble protein containing some enzymes. PMID- 12557660 TI - [Response and acclimation of photosynthesis in rice leaves to free-air CO2 enrichment (FACE)]. AB - The net photosynthetic rate (Pn), water use efficiency (WUE), and apparent quantum yield of carbon assimilation of rice leaves were boserved contrastively under ambient air (380 mumol.mol-1 CO2) and FACE (580 mumol.mol-1CO2). The results showed that the observed index under FACE were significantly higher than those under ambient air. Nevertheless, along with the time of high CO2 treatment prolonged, the enhancement effect of high CO2 on net photosynthetic rate declined gradually. At the same CO2 concentration, Pn and carboxylation efficiency (CE) in rice leaves under FACE were lower than those under ambient air. Although the stomatal conductances in FACE leaves was obviously lower than that in ambient leaves, their intercellular CO2 concentrations were not significantly different, which implied that the photosynthetic down-regulation in rice leaves grown under FACE was not caused by the decrease of stomatal conductance. PMID- 12557661 TI - [Effects of free-air CO2 enrichment (FACE) on yield formation in rice (Oryza sativa)]. AB - Effect of Free-Air CO2 enrichment (FACE, 200 mumol.mol-1 higher than CK) on rice yield and its components under field condition were studied with a Japonica variety 99-15. Results showed that FACE treatment had no obvious effect on plant height and number of leaves on main stem, but could quicken growth process of rice, shorten the whole growth duration. Increment of nitrogen application rate could weaken the effect of FACE on whole growth duration. The number of tillers and panicles, and fulfilled-grain percentage increased significantly under FACE treatment, whereas spikelets per panicle decreased. FACE treatment could significantly increase grain yield of rice, especially when higher rate of nitrogen fertilizer was applied. Increasing spikelets per panicle and spikelets per unit area under FACE treatment would result in marked increment of rice yield. Increasing nitrogen application rate was an important approach to increase spikelets per panicle and spikelets per unit area under FACE treatment. PMID- 12557662 TI - [Effect of free-air CO2 enrichment (FACE) on spikelets differentiation and retrogression in rice (Oryza sativa)]. AB - Effect of Free-Air CO2 Enrichment (FACE), 200 mumol.mol-1CO2 elevation under field condition, on the number of differentiated, retrograded, and survived primary and secondary branches and their spikelets in rice was investigated, and the percentage of retrograded branches and spikelets under FACE treatment were also calculated. Results showed that the number of differentiated primary and secondary branches, the number and percentage of retrograded primary branches per panicle were not significantly affected by FACE, but the number and percentage of retrograded secondary branches per panicle significantly increased under FACE treatment, which resulted in the significant decrease of survived secondary banches. FACE had no significant effect on the number of differentiated, retrograded and survived spikelets on the primary branches per panicle. The number of differentiated spikelets on secondary branches per panicle was not significantly affected by FACE, but FACE treatment significantly increased the number and percentage of retrograded spikelets on the secondary branches per panicle. Significant decrease of survived spikelets per panicle under FACE treatment was chiefly caused by marked increase of retrograded spikelets on the secondary branches of survived primary branches. FACE treatment significantly increased the surviving percentage of spikelets on the primary branches per panicle, while that on the secondary branches significantly decreased under FACE treatment. PMID- 12557663 TI - [Effect of free-air CO2 enrichment (FACE) on nitrogen accumulation and utilization efficiency in rice (Oryza sativa)]. AB - Effect of FACE, 200 mumol.mol-1 CO2 elevation under field condition in this study, on nitrogen accumulation and nitrogen use efficiency in rice was investigated. Results showed that nitrogen content (%) in rice plant sampled at different growth stages decreased significantly under FACE treatment, but FACE had no obvious effect on nitrogen accumulation in rice plant due to the significant enhancement in dry matter production. FACE treatment resulted in the significant increase in nitrogen use efficiency for biomass production (NUEp) which was measured at 28th day after transplanting, at heading and at maturity, respectively. Significant increase in nitrogen use efficiency for grain output (NUEg) and nitrogen harvest index (NHI) under FACE treatment was also observed in this study. Nitrogen content (%) and nitrogen accumulation in rice plant were increased under high N, but nitrogen use efficiency in rice plant were decreased. PMID- 12557664 TI - [Responses of rice (Oryza sativa) growth and its C, N and P composition to FACE (free-air carbon dioxide enrichment) and N, P fertilization]. AB - FACE (Free-air Carbon Dioxide Enrichment) was used to study the effects of elevated CO2 on rice (Oryza sativa) growth, tissue C/N, N and P concentration and uptake at different development stages under two N and two P levels. Results showed that elevated CO2 increased dry matter accumulation in rice stem, ear and root. Leaf dry matter was increased at tillering stage and no significant effect was found at jointing, heading and ripening stages. N concentration of stem and leaf was decreased. Ear N concentration at heading stage was increased but was decreased at ripening stage. No significant effect was found on root N concentration at tillering stage but root N concentration at jointing, heading and ripening was decreased. Leaf P concentration at jointing, heading and ripening was increased but no significant effect was found on P concentration in stem, ear and root. C content in various tissues changed unremarkably and the ratio of C over N (C/N) was increased. Elevated CO2 significantly increased P uptake in aboveground tissues; and increased N uptake, but the difference was not statistically significant. N and P fertilization had no significant effect on various tissue dry biomass. Tissue N content at higher N fertilization was higher than at lower N fertilization but no such effect of P fertilization on tissue P content was found. At higher N fertilization, elevated CO2 increased the ratio of below-ground biomass over above-ground biomass at ripening stage. Possible reasons are discussed for the differences of tissue N and P content and the ratio of below-ground biomass over above-ground biomass between elevated and ambient atmospheric CO2 concentrations. PMID- 12557665 TI - [Effect of FACE on competition between a C3 crop (rice, Oryza sativa) and a C4 weed (barnyardgrass, Echinochloa crusgalli)]. AB - The growth and competition of a C3 crop rice and a C4 weed, barnyardgrass under FACE(free-air carbon dioxide enrichment) conditions were studied by field experiment. The results showed that under FACE condition, the biomass and yield, leaf number, tiller number, and leaf area index of rice increased, while those of barnyardgrass decreased. Both of the leaf area indexed of rice and barnyardgrass were decreased, while their net assimilation rate increased. When the ratio of planting density of rice and barnyardgrass was 1:1, all of the ratio of biomass, yield, leaf area index, tiller and net assimilation rate were increased under FACE condition. It indicated that the competition between rice and barnyardgrass changed under FACE condition. The competition ability of rice(C3 crop) was enhanced relatively, while that of barnyardgrass weakened. PMID- 12557666 TI - [Effects of free-air CO2 enrichment on rice canopy microlimate]. AB - In this study, the free-air CO2 enrichment (FACE) system (setup at at Anzhen, Wuxi, Jiangsu Province in 2001) was used to investigate the effects of FACE on rice canopy microclimate. The rice canopy microclimate observations were carried out from August 26 to October 13, 2001, when the rice crops were at the heading to maturing development stage. The results showed that FACE reduced the rice leaf stomatal conductance. The rice leaf stomatal conductance difference between FACE and ambient was larger among upper layer leaves than among lower layer leaves and at heading and milk filling stages than at maturing stage. FACE increased daytime rice canopy temperature but had little effect on nighttime rice canopy temperature. The daytime rice canopy temperature difference between FACE and ambient was larger at heading and milk filling stages than at maturing stage. From heading to flowering, the daily maximum rice canopy temperature difference between FACE and ambient reached 1.2 degrees C under fine weather condition. The average daytime rice canopy temperature from flowering to maturing stage was about 0.43 degree C. Daytime air temperature inside rice canopy was also affected by FACE. Daytime air temperature inside rice canopy was higher in FACE plot than in ambient plot. The value of daytime air temperature difference between FACE and ambient increased with the increase of solar radiation and varied with height. The maximum daytime air temperature difference between FACE and ambient varied between 0.47-1.2 degrees C and 0.37-0.8 degree C at middle of canopy and canopy height, respectively. Air humidity and nighttime air temperature inside rice canopy were not significantly affected by FACE. These results indicate that FACE reducing rice leaf stomatal conductance was the major cause of the increase of canopy temperature and inside canopy air temperature in FACE plot. The higher canopy temperature and inside canopy air temperature in FACE plot resulted in the earlier maturity of rice crop in FACE plot than that in the ambient plot. PMID- 12557667 TI - [Determination of net exchange of CO2 between paddy fields and atmosphere with static poaque-chamber-based measurements]. AB - We firstly introduced the method for determining the net ecosystem exchange fluxes of CO2 (NEE) between croplands and atmosphere, based on field measurements using static opaquechamber/gas chromatography methods was introduced, and the application of this method in the FACE (free-air CO2 enrichment) study to examine the effects of elevated CO2 on the NEE over a typical paddy ecosystem was carried out, because of lacking in observation data for some necessary parameters, e.g., dark maintenance respiration coefficient, only the minimum value of NEE (NEEmin) was calculated based on opaque-chamber measurements. The NEEmin data indicate that CO2 elevated by 200 +/- 40 mumol.mol-1 significantly increased the ecosystem uptake of atmospheric CO2 by a factor ca. 3. To accurately determine the NEE based on opaquechamber measurements, dark maintenance respiration coefficient, above-ground biomass and root: shoot, i.e. R:S, ratio of root to shoot should be observed over the whole growing season. PMID- 12557668 TI - [Effects of elevated atmospheric CO2 on CH4 and N2O emissions from paddy fields]. AB - Effects of elevated atmospheric CO2 on CH4 and N2O emissions during the paddy rice-growing season were examined in a FACE (free-air carbon dioxide enrichment) study. The emission fluxes of CH4 and N2O from paddy rice fields were measured using methods based on static opaque-chamber and gas chromatography techniques. Synthetic fertilizer N was amended for the rice-growing season at two rates, 150 and 250 kgN.hm-2 and the atmospheric CO2 was enriched by 200 mumol.mol-1. At both N levels, the preliminary results indicate that no significant effect of CO2 enrichment on CH4 and N2O emissions from the rice paddy fields was detected. The result on CH4 emissions is inconsistent with the most literatures, and the result on N2O emissions is consistent with the most literatures. PMID- 12557669 TI - [Measurement of CO2 profiles in non-waterlogged soil in a FACE study]. AB - A method was specially designed and applied for measuring CO2 concentration of soil air over the non-waterlogged period of a rice-wheat rotation on an available area of about 1.6 m2 in a FACE (free-air CO2 enrichment) study. Based on measuring the CO2 concentration over the soil profile of 0-30 cm in depth using this method, the CO2 profile in the soils of wheat fields under elevated and ambient CO2 and the bare land under ambient CO2 was investigated and some preliminary results were obtained. Within 0-30 cm in soil depth, CO2 in the pores of the upper soil layers vertically diffused upwards much more quickly than that in the lower soil layers. During the period with active wheat growth, elevated atmospheric CO2 by 200 +/- 40 mumol.mol-1 significantly increased the CO2 concentration in soil air within 0-30 cm in depth by 14% +/- 5% (t-test, P < 0.001). PMID- 12557670 TI - [Rice-wheat rotational FACE platform. I. System structure and control]. AB - A Free Air CO2 Enrichment (FACE) system for rice and winter wheat rotation was established, elevated CO2 concentration was controlled to 200 mumol.mol-1 above ambient by computer system platform according to ambient CO2 concentration variation, wind direction, wind speed, canopy height and day-night changing. Experiments showed that the main factors affecting control precision are wind speed, crop and soil respiration and thickness of diffuse layer. After parameters adjustment, in daytime the time fraction for control precision achieve 80% is 83%, in night is 68%. The CO2 concentrations distribution in FACE rings are uniformity. The set CO2 is 557 mol.mol-1 in daytime and 608 mol.mol-1 in night. In 2001 rice season the target achievement ratios (TAR) were 1.03 after sunrise and 1.09 after sunset, respectively. PMID- 12557671 TI - [Rice-wheat rotational FACE platform. II. Data processing software package]. AB - A large amount data are brought from daily control, data collection and processing of FACE platform. A FACE system control, data collection and processing software package written in Visual BASIC including OLE to support office functions can finish the platform control, data collection, original data save and backup, daily data processing, monthly data processing and whole time FACE system control status analysis functions, so FACE system controllers can observe FACE system control status and keep control precision timely, and the researchers can get the data they want at any moment and convenient. PMID- 12557672 TI - [Field measurement of NO and NO2 exchanges between cultivated lands and the atmosphere in a FACE study]. AB - A method for measuring NO and NO2 exchanges between cultivated lands and the atmosphere in a FACE (free-air CO2 enrichment) study is described. With this method, gas is sampled with a technique of static-opaque-chamber and the fluxes of NO and NO2 exchanges are determined by analyzing the NO and NO2 concentrations with a chemiluminescent NOx analyzer. Application of this method in the FACE study of a rice-wheat ecosystem has indicated that reliable data on the exchange fluxes could be obtained. Over the non-waterlogged period of a rice-wheat rotation, net emission of NO from the fields was observed, while net uptake of NO2 occurred. The daily net emission of NO did not correlated with the soil temperature, but negatively depended upon soil moisture (R2 = 0.82, P < 0.001). A significant seasonal variation in the net uptake of NO2 was observed, which was regulated by wheat growth status. The daily uptake of NO2 depended upon both soil temperature and soil moisture. The dependence for each could be described with a parabola function (for soil temperature: R2 = 0.74, P < 0.001; for soil moisture: R2 = 0.69, P < 0.001). An elevation of atmospheric CO2 by 200 +/- 40 mumol.mol-1 mitigated the net emission of NO by 19% (t-test P = 0.096) and might be possible to reduce the net uptake of NO2 by 10% (t-test P = 0.26), which was likely due to the stimulated wheat growth. PMID- 12557673 TI - Effects of elevated atmospheric CO2 on nematode trophic groups in a Chinese paddy field ecosystem. AB - Soil fauna plays significant roles in the detritus food webs of agroecosystems, they are the essential contributors to the decomposition of soil organic matter, mineralization of plant nutrients and nutrient cycling. Some evidences indicate that soil fauna of the detritus food webs appears to show positive, neutral or negative responses to global change, especially atmospheric CO2 enrichment across different studies. Soil nematodes are representative of a large portion of this fauna, since they are abundant and trophically diverse in most soils. The free air CO2 enrichment (FACE) technology was used to expose three plots of rice (Oryza sativa) under elevated (ambient + 200 mumol.mol-1) atmospheric CO2, while three control plots were outfitted with FACE apparatus under ambient CO2. Nematode trophic groups were monitored in this study at depths of 0-5 cm and 5-10 cm during rice jointing and ripening stages in Wuxi site, China. Significant differences were found between depths and sampling dates in the total numbers of nematodes, bacterivores, plant parasites and omnivores-predators during the study period. The total numbers of nematodes and bacterivores at the 5-10 cm depth were higher in treatment plots than in control plots across all sampling dates. At 0-5 cm depth, the numbers of bacterivores were higher in treatment plots than in control plots, while those of omnivores-predators exhibited an inverse trend during the study period. Significant differences were also observed between treatment plots and control plots in the numbers of fungivores in our experiment. PMID- 12557674 TI - [Phase-directional management of protective plantations. II. Typical protective plantation: farmland shelterbelt]. AB - This paper is the second part of the series reports on phase-directional management of protective plantations. Based on the fundamentals of phase directional management for protective plantations in paper I, discussions on protective maturity, management phase, and regeneration patterns and methods for shelterbelt or windbreak were conducted. Firstly, the two points of protective maturity of Populus spp. i.e., initial protective maturity age (IPMA) and terminal protective maturity age (TPMA), were determined. According to the growth pattern of shelterbelt, the IPMA was 16 years and 24 years for local poplar and hybrid poplar shelterbelts, respectively, and TPMA was the natural age for both local and hybrid poplar shelterbelts, which was determined on the basis of change pattern of porosity. Secondly, the three management phases of shelterbelts were divided quantitatively based on the analyses of discrete degrees of diameter, IPMA and TPMA. Thirdly, the corresponding measures for phase-directional management in each phase were also developed. In pre-maturity phase, i.e., the period from establishment of shelterbelt to IPMA, the measures such as soil cultivation, irrigation, fertilization, weed clearing, intercropping and branch cutting etc. should be conducted. In maturity phase, i.e., the period from IPMA to TPMA in theory, but the period from IPMA to regeneration age in practice, the techniques of managing the shelterbelt porosity around 0.25 (the best porosity value for wind protection) should be developed. In this phase, the porosity model was constructed according to the distribution pattern of base area for the poplar shelterbelt. The classification of shelterbelt trees, i.e., 4-grade classification, was established by considering the contribution of trees to the shelterbelt structure, which can provide the standard for thinning and tending of shelterbelt. Additionally, the density controlling techniques were also discussed. In the period of regeneration, i.e., from the regeneration age to the establishment of next generation shelterbelt, the techniques of regeneration such as determination of regeneration age, arrangement of regeneration patterns should be paid great emphasis. The influences of regeneration patterns and methods on regeneration period were specially discussed, and the changes of three management phases were illustrated by different regeneration patterns in the end of this paper. These results were obtained from poplar shelterbelt, but the fundamentals could be applied in any other shelterbelts. PMID- 12557675 TI - [Effects of enhanced ultraviolet-B radiation on growth development and yield formation in rice (Oryza sativa L.)]. AB - Effect of enhanced ultraviolet-B radiation (280-320 nm) on growth, development and yield formation in the three rice cultivars (Oryza sativa L.) under pot conditions were investigated. The results showed that enhanced ultraviolet-B radiation decreased rice height, tiller number, total leaf area and biomass, but the inhibitory rate varied in different cultivars and developmental stages. Plant height was decreased by 9.4%-12.2% in seedling stage, plant shoot and root of dry matter were decreased by 45.3%-59.8% and 54.9%-59.0% respectively in tillering period. The leaf emergence delayed, resulting in delayed time of flowering in Shanyou 63, Nanchuan and IR65600-85 by 2d, 3d and 7d, prolonged total growth period by 3d, 4d and 9d. Net photosynthesis rate of Shanyou 63, Nanchuan and IR65600-85 were decreased by 11.9%, 12.8% and 29.7%, respectively. Attributed to decreased contents of chlorophyll a, b, carotenoid and induction kinetics parameters of Fv, Fv/Fm and Fv/Fo. Effective panicle per plant, total grain number per panicle, filled grain rate and 1000-grain weight were decreased by enhanced UV-B radiation. Consequently the grain yields were reduced by 25.2% 31.1%. PMID- 12557676 TI - [Principal axes analyses on population genetic differentiation of Plantago asiatica in Zhejiang]. AB - Eight populations of Plantago asiatica collected from Zhejiang were analyzed using random amplified polymorphic DNA (RAPD). Eighteen of the 60 random primers were selected. 186 bands were amplified by the 18 primers, and 68.70% of them were polymorphic. Jaccard coefficient was applied to compare the genetic of these eight populations based on the RAPD data. Three-dimensional ordination plot of the eight populations based on the Jaccard coefficient was constructed by using principal axes analysis (PAA). The results showed that the genetic differentiation of eight populations could be explained to a certain extent by their geographical location and altitude. PMID- 12557677 TI - [Effects of plant foliar litter diversity on the growth of Chinese fir seedling and the absorption of (15NH4)2SO4]. AB - Effects of plant foliar litter species composition on the growth of the potted Chinese fir seedling and the absorption of (15NH4)2SO4 was studied. The result showed that there were significant growth responses to foliar litter treatments. A very noticeable trend was that as the tree species of foliar litter placed on pots increases, the growth of Chinese fir seedlings increases steadily. Seedling growth in four-species mixed foliar litter were the largest and that in Cunninghamia lanceolata + Michelia macclurei + Kalooanax septemlobus foliar litter mixture were the second. The others were in order of seedlings in C. lanceolata + M. macclurei + Castanopsis carlesii foliar litter mixture > C. lanceolata + K. septemlobus foliar litter mixture > C. lanceolata + C. carlesii > control (no foliar litter) > C. lanceolata + M. macclurei foliar litter mixture > Chinese fir foliar litter treatment. The recovery of (15NH4)2SO4 by Chinese fir seedlings in no foliar litter treatment was maximum, and seedlings grown in four foliar litter mixture was the second, then it was in order of C. lanceolata + C. carlesii + K. septemlobus = C. lanceolata + C. carlesii + M. macclurei > C. lanceolata + K. septemlobus > C. lanceolata + C. carlesii > C. lanceolata + M. macclurei > C. lanceolata foliar litter treatment. In addition, the residue of (15NH4)2SO4 was greater in soil with foliar litter treatment than in no foliar litter soil. Both of the residual amount of (15NH4)2SO4 in soil and the total recovery of (15NH4)2SO4 increased with the increasing tree species of applied foliar litter. PMID- 12557678 TI - [Effect of vanillin and P-hydroxybenzoic acid on physiological characteristics of Chinese fir seedlings]. AB - Effects of vanillin and P-hydroxybenzoic acid at different concentrations on physiological characteristic of Chinese fir seedlings were studied by potted experiment. The results showed that 10 mmol.L-1 and 1 mmol.L-1 of two kinds of phenolics significantly reduced the content of chlorophyll, rate of photosynthesis and root activity, and that the higher the concentration of vanillin and P-hydroxybenzoic acid, the more the physiological activities was inhibited. By treatment with 1 and 10 mmol.L-1 vanillin, Chinese fir seedlings reduced its photosynthesis rate 25.1% and 37.0%, transpiration rate 20.3% and 37.0%, stomata conductance 33.7% and 46.8% and root activity 51.6% and 78.8%, respectively. The results suggested that vanillin and P-hydroxybenzoic acid accumulated in the soil by continuous cropping of Chinese fir may have some allelopathic effect on the seedlings of Chinese fir and the effect is one of the factors leading to the low productivity of continuously cropped Chinese fir forest. PMID- 12557679 TI - [Identification on decomposing products of soybean stubs by Trichodermar koningii and their allelopathy]. AB - Different solvents including cyclohexane, ethyl acetate, n-butanol and XAD-4 resin were used to extract the allelochemicals from the decomposing products of soybean stubs. The results showed that the germination of soybean seeds were inhibited at initial stages by the solution extracted with cyclohexane, ethyl acetate, n-butanol and resin from the decomposing products of soybean stubs that had been incubated for 2 and 4 weeks. The inhibition was gradually weakened with the time. When concentrations of the ethyl acetate and resin fractions were 0.50, 1.00 gDW.ml-1 and when their concentrations were 0.10, 0.25 gDW.ml-1, their RI (the index of allelopathic effect) value of soybean radicle length reached significant level. The RI values of the former and those of the cyclohexane fractions reached significant level also. The chemical constituents of allelopathic fractions were identified by GC-MS as organic acids, phenol, aldehyde, ketone, benzene, amine, nitrile, ester and some other compounds, some of which were allelochemicals. PMID- 12557680 TI - [Allelopathic potential of volatile oil from Mikania micrantha]. AB - Mikania micrantha, an exotic plant species, is an important weed in south China. The volatile oil of the weed species had significant biological activities on plants and their pathogens, particularly Pyricularia grisea. The results showed that the seeding growth of 6 plant species decreased obviously when exposed to increasing concentration (200, 400, 800, 1600 mg.L-1) of the volatile oil. The fresh weight of all test plants decreased, and the emergence of all test plants delayed for 1-2 days under soil treatment (2500 g.hm-2). The inhibitory effect of volatile oil of M. micrantha at its medium concentration was the most strong on Pyricularia grisea; secondary on Fusarium oxysporum; but weak on Phytophthora nicotianae. The inhibitory rate was 53.38%, 28.66% and 18.69% for the three plant pathogens, respectively. PMID- 12557681 TI - [Effects of NaCl stress on seedling growth and IAA metabolism of sweet potato and its relation to salt-tolerance]. AB - Seedlings of three sweet-potato (Ipomoea batatas) cultivars with different salt tolerance were stressed with different concentrations of NaCl during the seedling stage. Plant growth, leaf area growth, and IAA content decreased with increasing NaCl concentration, while the activities of POD and IAA oxidase increased. The extent of plant growth, leaf area growth and IAA content decrease and the extent of POD and IAA oxidase activity increase were larger in sensitive cultivar than those in moderately tolerant and tolerant cultivars. In addition, the content of chlorogenic acid increased under NaCl stress. Chlorogenic acid content reached its peak value at 170 mmol.L-1 of NaCl in sensitive and moderately tolerant cultivars and 225 mmol.L-1 of NaCl in tolerant cultivar. Chlorogenic acid content decreased with increasing NaCl concentration, but it was higher than that of the control. PMID- 12557682 TI - [Geostatiscal analysis on spatial patterns of Schizaphis granarium and Aphidius sp]. AB - The spatial construction and correlation of Schizaphis granarium and Aphidius sp. at different stages were investigated using geostatistical method. The results showed that the semivariogram of S. granarium was described by a spherical model, and indicated an aggregated special pattern ranging from 21 to 61 cm. The semivariogram of Aphidius sp. could be also well described by spherical model, and indicated an aggregated spatial pattern ranging from 36 to 55 cm. The amount and spatial distribution of Aphidius sp. were closely related to those of Schizaphis granarium, which reveals that Aphidius sp. was the dominant natural enemies influencing the population of S. granarium. PMID- 12557683 TI - [Relationship between climatic factors and wannanhua sow's reproductivity]. AB - Data on reproductive traits of 670 litters of different Am genotypes Wannanhua sow under different climatic conditions were analyzed by the least square model in order to investigate the effects of the climate factors and the Am genotypes on sow's reproductivity. The results showed that the reproductive performance of sow depended on the monthly change of climate regularly. High temperature from July to September had worse effect on sow's reproductivity, especially (P < 0.05). A certain variation existed among reproductive performances of Wannanhua sows at Am loci. AmAB's was better, followed by AmAA, then AmBB and AmBC were poor. But the variance was in significant (P > 0.05). Furthermore, there was strong interactive effect between Am genotypes and the breeding season. In addition, the litter size, the number of live born and the number of weaning pig had a negative correlation with the mean air temperature and photoperiod significantly (P < 0.05 or P < 0.01). PMID- 12557684 TI - [Lake ecosystem services and their ecological valuation--a case study of Baoan Lake in Hubei province]. AB - Along with the maturation of estimation methods for biological resources, the studies on the services value of ecosystem become a hotspot in ecology. The studies on the services value of lake ecosystem were not been reported before this issue, and this issue firstly attempted to estimate the Baoan Lake ecosystem services and its indirect economic values on the basis of ecological function analysis and some economic methods. The results showed that the indirect economic value (in RMB) of organic matter production, CO2 fixation, O2 release, nutrient recycling, water conservancy and water supply and SO2 degradation were 1.51 x 10(7), 4.77 x 10(7), 3.63 x 10(7), 4.45 x 10(5), 1.10 x 10(8), 5.15 x 10(5) yuan.yr-1, respectively. The sum was about 2.24 x 10(8) yuan.yr-1. From the above incomplete estimation, it was known that the Baoan Lake ecosystem has great ecological and economic values. PMID- 12557685 TI - [Effect of sewage irrigation on wheat growth and its activate oxygen metabolism]. AB - The effects of sewage irrigation on the seedling growth of wheat and its activate oxygen metabolism were studied by water cultivation experiments. The results showed that compared with control, the wheat under sewage irrigation was short and small, and its roots number, fresh/dry weight of stem or root, and content of soluble protein decreased drastically. The content of chlorophyll decreased, while the Cyta/Cytb and membrane lipid peroxidation were increased. The sensitivity of pigment content of wheat leaf undr sewage irrigation was in order of Chlorophyll b > Chlorophyll a > Carotenoid. Furthermore, the POD, SOD and CAT activities of the wheat went down in varying degrees, and the SOD was the most susceptible enzyme to waste water stress. In this paper, the activities of POD, SOD and CAT decreased in 19.1%-33.1%, 37.8%-47.8% and 30.2%-35.9%, respectively. PMID- 12557686 TI - [Responses of agricultural crops of free-air CO2 enrichment]. AB - Over the past decade, free-air CO2 enrichment (FACE) experiments have been conducted on several agricultural crops: wheat(Triticum aestivum L.), perennial ryegrass (Lolium perenne), and rice(Oryza sativa L.) which are C3 grasses; sorghum (Sorghum bicolor (L.) Moench), a C4 grass; white clover (Trifolium repens), a C3 legume; potato (Solanum tuberosum L.), a C3 forb with tuber storage; and cotton (Gossypium hirsutum L.) and grape (Vitis vinifera L.) which are C3 woody perennials. Using reports from these experiments, the relative responses of these crops was discussed with regard to photosynthesis, stomatal conductance, canopy temperature, water use, water potential, leaf area index, shoot and root biomass accumulation, agricultural yield, radiation use efficiency, specific leaf area, tissue nitrogen concentration, nitrogen yield, carbohydrate concentration, phenology, soil microbiology, soil respiration, trace gas emissions, and soil carbon sequestration. Generally, the magnitude of these responses varied with the functional type of plant and with the soil nitrogen and water status. As expected, the elevated CO2 increased photosynthesis and biomass production and yield substantially in C3 species, but little in C4, and it decreased stomatal conductance and transpiration in both C3 and C4 species and greatly improved water-use efficiency in all the crops. Growth stimulations were as large or larger under water-stress compared to well-watered conditions. Growth stimulations of non-legumes were reduced at low soil nitrogen, whereas elevated CO2 strongly stimulated the growth of the clover legume both at ample and under low N conditions. Roots were generally stimulated more than shoots. Woody perennials had larger growth responses to elevated CO2, while at the same time, their reductions in stomatal conductance were smaller. Tissue nitrogen concentrations went down while carbohydrate and some other carbon-based compounds went up due to elevated CO2, with leaves and foliage affected more than other organs. Phenology was accelerated slightly in most but not all species. Elevated CO2 affected some soil microbes greatly but not others, yet overall activity appears to be stimulated. Detection of statistically significant changes in soil organic carbon in any one study was impossible, yet combining results from several sites and years, it appears that elevated CO2 did increase sequestration of soil carbon. Whenever possible, comparisons were made between the FACE results and those from prior chamber-based experiments reviewed in the literature. Over all the data and parameters considered in this review, there are only two parameters for which the FACE- and chamber-based data appear to be inconsistent. One is that elevated CO2 from FACE appears to reduce stomatal conductance about one and a half times more than observed in prior chamber experiments. Similarly, elevated CO2 appears to have stimulated root growth relatively more than shoot growth under FACE conditions compared to chamber conditions. Nevertheless, for the most part, the FACE- and chamber-based results have been consistent, which gives confidence that conclusions drawn from both types of data are accurate. However, the more realistic FACE environment and the larger plot size have enabled more extensive robust multidisciplinary data sets to be obtained under conditions representative of open fields in the future high-CO2 world. PMID- 12557687 TI - [Elevated atmospheric CO2 and crop/weed competition]. AB - Elevated atmospheric CO2 has direct photosynthesis effects on photosynthesis, respiration and water use efficiency, as well as on the growth and reproduction of plants. CO2 enrichment is more beneficial to plants with the C3 photosynthetic pathway than to those with the C4 photosynthetic pathway. Most major weeds are C4 plants, while many crops are C3 plant. Therefore, different effects of elevated atmospheric CO2 have important implications for weed/crop interaction. In this paper, the effect of CO2 enrichment and global chang on weed/crop interaction were summarized, and some theoretical and practical aspects were discussed. PMID- 12557689 TI - [Current situation and prospect of elevated atmospheric CO2 effects on soil nematodes]. AB - The elevated atmospheric CO2 can make effects on the ecosystem, these effects will be mediated by the soil fauna. In this paper, we firstly discussed the response of the soil nematodes to the elevated atmospheric CO2 in different soil pattern and in different ecosystems. Some evidences indicate that soil fauna of the detritus food webs appear to show positive, neutral or negative responses to the atmospheric CO2 enrichment across different studies. The research prospect in the future was put forward. This paper can help us understand some ecological processes of farmland ecosystems according to the effect of elevated CO2 on soil nematodes and provide the foundation for managing farmland ecosystems effectively. PMID- 12557688 TI - [Effects of elevated atmospheric CO2 on the exchange of trace gases between ecosystems and the atmosphere]. AB - The latest researches on the effects of elevated atmospheric CO2 on the exchanges of trace gases (e.g., CO2, CH4 and N2O) between the atmosphere and ecosystems were reviewed. The techniques and methods involved in the researches were introduced firstly. Then the review mainly focused upon the results from those studies using the open-top-chamber (OTC) methods and the free-air carbon dioxide enrichment (FACE) system. Generally, elevated atmospheric CO2 may stimulate biomass accumulation, and enlarge C/N ratio in plant tissue so as to reduce the decomposition of organic matter. This action could increase CO2 sequestration in terrestrial ecosystems. Elevated atmospheric CO2 could impact on methanogenic bacteria and CH4 emissions. An increase in CH4 emissions from wetland may appear. The argument among the responses of N2O emissions to elevated CO2, however, was inconsistent. So far, no study on other trace gases was reported. More efforts should be taken in the research on the effects of elevated atmospheric CO2 on the exchange of trace gases. PMID- 12557690 TI - [Phase-directional management of protective plantations. I. Fundamentals]. AB - In order to ensure the higher effectiveness, more stability and sustainability of Protective plantations, the definition of phase-directional management (PDM) of protective plantations was brought forward on the basis of management researches and practices for protective plantations. The basics of PDM is protective maturity, which is defined as the time when the protective plantations attain to the state that the protective plantations can provide effective and complete protection to the objects needed to be protected. Protective maturity has two points, initial protective maturity age (IPMA), the age of protective maturity started, and terminal protective maturity age (TPMA), the age of protective maturity ended. Three management phases of protective plantation, i.e., prematurity phase, the period from sapling or establishment to initial protective maturity, protective maturity phase, the period of protective maturity lasting, and regeneration phase, the period during regeneration and before the establishment, are divided based on the fundamental of protective maturity. Directional management of protective plantation means that all of the management techniques in each phase are directed at the aim of protective maturity, i.e., protective maturity is the direction of management of protective forests, and protective maturity is the final objective for the management of protective forests. In order to sustain the protective maturity state, corresponding measures should be conducted in each phase, according to the classification of protective plantations. In pre-maturity phase, the purpose of managing is to accelerate the protective maturity, therefore, the measures such as weed clearing, soil cultivation, irrigation, fertilization, intercropping and branch cutting etc. should be conducted in protective plantations. In maturity phase, the aim of managing is to sustain the protective maturity, i.e., the techniques (tending and thinning) of controlling the structure of protective plantations should be paid emphases. In the period of regeneration, the objective of managing is to recover the protective maturity, accordingly, the regeneration patterns and ways should be determined reasonably. Additionally, the methods of determining protective maturity, i.e., the core of the phase-directional management, are also given corresponding to farmland shelterbelt, sand-fixation forest and water and soil conservation forest. For farmland shelterbelt, IPMA can be determined according to the growth pattern of tree height under the suitable structure (porosity). For sand-fixation forest, it can be determined by the cover degree of the forest, i.e., the age when cover degree gets to pi/4 can be considered as IPMA. In the case of water and soil conservation forest, IPMA is determined by the canopy closure at the height of 1m above forest ground, which can be obtained from the hemispherical silhouettes in vertical direction. As for the TPMA, it can be described by the natural age of trees for all of above-mentioned three kinds of protective plantations, but the concrete methods for estimating the natural age of trees in each kind of forests are different. PMID- 12557691 TI - [Effect of elevated atmospheric CO2 on soil urease and phosphatase activities]. AB - The response of soil urease and phosphatase activities at different rice growth stages to free air CO2 enrichment (FACE) was studied. The results showed that comparing with the ambient atmospheric CO2 concentration (370 mumol.mol-1), FACE (570 mumol.mol-1) significantly increased the urease activity of 0-5 cm soil layer at the vigorous growth stage of rice, whole that of 5-10 cm layer had no significant change during the whole growing season. Phosphatase activity of 0-5 cm and 5-10 cm soil layers significantly increased, and the peak increment was at the vigorous growth stage of rice. PMID- 12557692 TI - [Effect of free-air CO2 enrichment on soil microbe in paddy field]. AB - Utilizing FACE research site in Anzhen of Wuxi, the effect of CO2 concentration increasing on soil microbe was studied in a paddy field under the condition of local cultivation, irrigation and fertilization. The results showed that soil respiration intensity was 31.88 ml.kg-1 and bacteria number was 1.51 x 10(6) ind.g-1 dry soil in FACE area, which were greater than those in control area (soil respiration intensity 27.83 ml.kg-1, bacteria number was 1.25 x 10(6) ind.g 1 dry soil, but the differences were not significant. PMID- 12557693 TI - Darwin, Cuvier and Geoffroy: comments and questions. AB - In writing, in the Origin of Species, of 'two great laws' on which organic beings are formed, 'Unity of Type' and 'Conditions of Existence', Darwin was referring to the famous opposition between Cuvier and Geoffroy Saint-Hilaire, first stated publicly in the spring of 1830. After a brief statement of the chief points at issue in the debate, I raise the question of Darwin's attitude to the disagreement and the views of the two protagonists. There are numerous earlier, and some later, references to Cuvier and Geoffroy in the Darwin archives, notebooks, marginalia and correspondence. An examination of these materials suggests a shift in Darwin's sympathies, from Geoffroy to Cuvier. However, some of Geoffroy's principles are retained, and, in adopting Cuvier's phrase 'conditions of existence', Darwin partly alters its meaning. Finally, since originally, and in its adoption by such writers as Whewell and Owen, the expression 'conditions of existence' was interpreted as entailing design or final cause, I consider the vexed question of Darwin's attitude to teleology. PMID- 12557694 TI - [Humboldt as mediator: Schleiden and Mohl contra Liebig]. AB - Justus Liebig's book Die organische Chemic in ihrer Anwendung auf Agricultur und Physiologie aroused strong opposition from scientists who felt that they were being attacked. A polarization arose particularly between Hugo von Mohl (1805 1872) and Matthias Jacob Schleiden (1805-1881) on one side, and Justus von Liebig (1803-1873) and his followers on the other side. The debate did not have the character of a purely scientific controversy, however, because social aspects also played a role. Alexander von Humboldt (1769-1859), who held the esteem of both parties, was able to intervene successfully to mediate this conflict. His task appeared at first difficult, but it turned out that both groups had a common enemy, the representatives and followers of the 'Romantic philosophy of nature'. Humboldt, who self-deprecatingly called himself a 'fossil', was forced to recognize that many of his friends and acquaintances, who stood nearer to, or even belonged among the founders of Naturphilosophie, for example, Friedrich Wilhelm Joseph von Schelling (1775-1854) and Johann Bernhard Wilbrand (1779 1846), were also under attack. Although he had earlier been inspired by the concerns and ideas of Schelling's Naturphilosophie, he now distanced himself from them in order to mediate between modern, experimental scientists. PMID- 12557695 TI - [The research on the premature brain as a multidisciplinary model at the Baudelocque de Paris Clinic, 1942-1962]. AB - Studies on the premature brain started in France in the Baudelocque clinic in Paris. It was the opportunity for a new clinical and epistemological paradigm: the pluridisciplinary model. The need to save and prevent premature babies led the founder of this research, Alexander Minkowski, to gather together, in the same laboratory, biochemistry, anatomo-pathology, electrophysiology and developmental science. The models of the premature brain helped establish a scale of biological maturation, the degrees of neurobiological development, and the electric intensity of mental states. PMID- 12557696 TI - Can biologists and philosophers see eye to eye on function? Essay review. PMID- 12557697 TI - [The new mechanisms and reversal of drug resistance]. AB - The emergence of tumors resistant to anticancer agents has been recognized as one of the major obstacles to effective cancer chemotherapy. Recent studies have elucidated various mechanisms in this resistance, such as induction of drug efflux pumps, in particular certain of the ABC transporters. Experiments to predict the sensitivity of tumors to anticancer agents using DNA microarray and SNPs analyses are ongoing. The development of the new anticancer agent ST1571 and the emergence of tumors resistant to ST1571 indicate the urgent need for clinically available reversing agents. PMID- 12557698 TI - [Prostate-specific antigen and its related parameters in detecting prostate cancer]. AB - Prostate-specific antigen (PSA) is the most useful tumor marker available today. However, its inability to clearly distinguish between prostate cancer and benign prostatic hyperplasia may result in serial and unnecessary prostate biopsies. Various attempts have been made to improve its specificity. PSA-related parameters including PSA density, PSA density for the transition zone, age- or race-specific PSA reference range, PSA velocity, and percent free PSA are the most promising approaches available at present. PMID- 12557699 TI - [Clinical problems of systematic prostate needle biopsy in the detection of prostate cancer]. AB - Prostate needle biopsy is necessary for the pathological diagnosis of prostate cancer. Currently, sextant systemic random biopsy under transrectal ultrasound is widely used. However, several clinical problems with this biopsy have been discussed. This review focuses on these clinical points in prostate biopsy. PMID- 12557700 TI - [Problems of the preoperative prediction of the pathological stage in prostate cancer]. AB - The pathological stage of the tumor is the most influential prognostic factor for progression after radical prostatectomy. However, as many as 50% of men undergoing radical prostatectomy are found to have extraprostatic disease in the pathological specimen. Accurate identification of the risks of disease extension and of disease recurrence prior to radical prostatectomy would thus be useful in counseling men presenting with clinically localized prostate cancer. Nomograms may help patients and physicians make more informed treatment decisions based on the probability of pathological stage. Partin and co-workers popularized the use of a pretreatment nomogram based on PSA (prostate specific antigen), clinical stage (TNM stage) and biopsy Gleason score to predict the pathological stage of localized prostate cancer. However, it may not be directly applicable to Japanese males, and the interpretation and comparison of data sets should be done with caution and careful consideration. Although attempts have been made to establish a nomogram for Japanese patients, been tried, it is still based on the data for a small number of patients. More data from a greater number of patients and validation analysis are essential. Recently, artificial neural networks (ANN) have been shown to be effective in predicting pathologic stage in men with clinically localized prostate cancer. The use of ANNs is a relatively new concept and the data is based on Western people; thus, the data analysis for Japanese patients is necessary. The present paper mainly outlines the usefulness and problems for the preoperative prediction of the pathological stage in prostate cancer by nomograms and artificial neural networks. PMID- 12557701 TI - [Treatment strategy of localized prostate cancer]. AB - In recent years, early detection techniques including PSA have resulted in a dramatic increase in the number of patients with localized prostate cancer. The natural history of localized prostate cancer is unusually favorable as compared with many other common cancers. Because of this, the management is controversial. Radical prostatectomy (RP), various techniques of irradiation (RT), endocrine therapy (ET) and watchful waiting (WW) have been utilized, but well designed randomized clinical trials with end results are lacking. Currently, RP is generally accepted as the preferred treatment option for younger patients with lower PSA level. RT has been thought to be somewhat inferior in terms of long term survival rate compared with RP. Aggressive treatments such as RP and RT are associated with side effects: RP has more urinary and sexual side effects, while RT has more bowel symptoms. For some patients of older age, with well differentiated tumor and lower PSA level, WW is recommended. WW is devoid of treatment-related side effects, but patients have the risk of progression. Several clinical trials of treatment for localized prostate cancer are being conducted in Western countries. However, these studies will require at least 10 years to accumulate enough study endpoints to form any conclusion. Therefore, it is important for the treatment strategies that we should carry out interventions that are neither excessive nor insufficient for each patient, and take QOL into consideration. PMID- 12557702 TI - [Therapeutic strategy for prostate specific antigen (PSA) failure after radical prostatectomy]. AB - Prostate specific antigen (PSA) failure occurs in 20% to 50% of patients who undergo radical prostatectomy. There is no consensus on how PSA failure should be managed. Recently, salvage radiotherapy is reported to be an effective treatment for PSA failure, and 20%-70% of cases are recurrence-free. The aim of salvage radiotherapy is a cure and this is quite different from other options, for example, endocrine therapy. Salvage radiotherapy is likely to become more important as a treatment for PSA failure after radical prostatectomy. PMID- 12557703 TI - [Treatment strategy for locally advanced prostate cancer]. AB - With the increasing aged population, prostate cancer has become one of the commonest malignant tumors in the United States. The incidence of prostate cancer is the highest among malignant tumors in males, and the mortality rate is the second highest following lung cancer. Even when prostate cancer is diagnosed to be in the early stage preoperatively, its excised lesions are often judged pathohistologically to be locally advanced tumor (staging error). Therefore, to estimate the exact pathological stage of excised lesions by preoperative parameters such as clinical T, PSA and biopsy Gleason Score, Partin's nomogram is generally used in the United States. However, according to the annual update version of the 2001 millennium update, radical prostatectomy should not be applied to T3, and it was excluded from the nomogram. Currently, the standard methods for the treatment of locally advanced prostate cancer may be external beam radiotherapy and brachytherapy with neoadjuvant hormonal therapy and intraprostate 125I and 103Pd seeds with neoadjuvant hormonal therapy, although the long-term results are unknown. In our study, similar to a report by Messing et al., adjuvant hormonal therapy might be effective in patients in whom the tumor was diagnosed as being in the early stage but was later found to be N (+) after its operation. PMID- 12557704 TI - [First line therapy in the treatment of metastatic prostate cancer]. AB - Hormonal therapy has been the main treatment for advanced prostate cancer for the past six decades. Maximum androgen blockade (MAB), combination therapy with castration and antiandrogens, has been compared with castration monotherapy since the late 1980s. However, the results of the different trials have been conflicting. Recently published meta-analyses have revealed that MAB with non steroidal antiandrogens is slightly superior to monotherapy (surgical or medical castration) in the treatment of advanced prostate cancer, and that MAB has more adverse effects. The question of whether the modest survival advantage of MAB balances with the increase in adverse effects should be investigated. Some studies indicated that the survival of metastatic prostate cancer patients treated with immediate therapy was similar to that of men in whom treatment was delayed. Hormonal therapy has side effects and is costly, and delayed treatment may be beneficial to elderly men with silent metastasis. Intermittent hormonal therapy is a controversial approach to management of advanced prostate cancer, although laboratory data suggest that intermittent androgen deprivation may prolong the duration of androgen dependence. Intermittent hormonal therapy for patients with metastatic prostate cancer needs to be assessed in a randomized trial to determine the effect on overall survival and quality of life. Our results in a randomized clinical trial of chemo-endocrine therapy versus endocrine therapy alone suggested that the addition of chemotherapy (cisplatin plus pirarubicin) to initial endocrine therapy might be beneficial to patients with advanced prostate cancer, especially an aggressive form of prostate cancer. However, chemo-endocrine therapy should be considered an experimental approach at present. PMID- 12557705 TI - [Evaluation of the increasing serum lactate dehydrogenase caused by recombinant human granulocyte-colony stimulating factor]. AB - OBJECTIVE: Increasing serum lactate dehydrogenase (LDH) is often caused by granulocyte-colony stimulating factor (G-CSF) for leukopenia following chemotherapy in patients with lung cancer. To evaluate the increase in LDH, we investigated the significance of its elevation and LDH isozyme during chemotherapy supported by recombinant human G-CSF (rhG-CSF). STUDY DESIGN: To exclude effects of liver diseases and chemotherapy-induced liver dysfunction, only patients in whom laboratory findings concerning liver function were within normal range were entered in this study. If leukocyte or neutrophil counts were less than grade 3, subcutaneous injection of 50 micrograms/m2 of filgrastim was given daily until leukocyte counts increased to more than 10,000/mm3. RESULTS: Sixty patients with unresectable lung cancer were enrolled in this study and the LDH isozyme was evaluable in 54 patients. Increasing LDH was observed in 38 patients(70.4%), and LDH isozyme was measured in these 38 patients. Increases in granulocytes and LDH isozymes were found to have a positive correlation. LDH2, LDH3, LDH4 and LDH5 increased significantly after rhG-CSF administration, although LDH 1 did not increase. CONCLUSION: It was found that a rapid increase in leukocytes by rhG-CSF induced an increase in LDH, especially LDH 3.4. Considering the results of principal component analysis and the distribution ratio of LDH isozymes in neutrophils, it is thought that elevation of LDH is reflected in the rapid production and consumption of neutrophils. PMID- 12557706 TI - [Cisplatin, 5-fluorouracil, and high-dose leucovorin for advanced esophageal cancer]. AB - We report the effects and toxicities of intravenous administration of cisplatin, 5-FU and high-dose leucovorin for advanced esophageal cancer. Eight patients were registered and sixteen lesions were measurable. Of these sixteen lesions, thirteen were primary or synchronous metastatic lesions, and the response was 69%. Three were recurrence lesions, and the response rate for them was 0%. Including seven neoadjuvant cases, ten patients had oral mucositis, and seven patients had appetite loss. Other toxicities were diarrhea, myelosuppression, renal dysfunction, and alopecia. All were reversible after administration. It is suggested that this treatment regimen is a superior neoadjuvant chemotherapy with low toxicity. PMID- 12557707 TI - [Weekly administration of paclitaxel with a short course of premedication for advanced or recurrent gastric cancer]. AB - Weekly administration of paclitaxel with a short course of premedication was performed for 8 patients with advanced or recurrent gastric cancer. In this regimen, 500 ml of physiological saline with vitamins was administered in a 3 hour infusion. After 30 minutes of infusion, dexamethasone 10 mg, chlorpheniramine maleate 5 mg, famotidine 20 mg and ramosetron hydrochloride 0.3 mg were administered intravenously. After 30 more minutes of infusion, paclitaxel at a dose of 65 mg/m2 was admixed in the residual normal physiological saline and administered over 2 hours. Administration was continued for 3 weeks with a 1 week rest. Though the partial response rate was 25%, clinical symptoms improved in all patients. Moreover, both hematological and non-hematological toxicities were mild. Weekly administration of paclitaxel with a short course of premedication is an effective and well-tolerated method for patients with advanced or recurrent gastric cancer. PMID- 12557708 TI - [Pharmacoeconomic study of chemotherapy for gastric cancer: analysis of medical costs for oral fluoropyrimidine TS-1 and conventional i.v therapy]. AB - To evaluate the economic impact of TS-1, an oral fluoropyrimidine, on the treatment of gastric cancer, the medical costs required for TS-1 treatment were compared with those for the conventional chemotherapy employed before the launch of TS-1 in patients with advanced and recurrent gastric cancer. The medical costs for 13 patients receiving TS-1 and 10 patients undergoing the conventional chemotherapy were extracted from the ordering system data, and the costs were compared using the fee schedule of the Japanese national health insurance. The monthly medical costs for the TS-1 group and conventional chemotherapy group were 327, 640 +/- 47,647 (mean +/- SE) yen and 852,874 +/- 62,412 yen, respectively. Medical costs appeared to have decreased because TS-1 is an oral preparation, permitting an easy transfer from inpatient treatment to ambulatory treatment, and because only small amounts of medication and blood transfusion were used for supportive care. Consequently, the medical costs for the TS-1 group were significantly lower than for the conventional chemotherapy group. Therefore, the administration of TS-1 leads to a reduction in medical costs. PMID- 12557709 TI - [Early effect of intra-arterial chemotherapy combined with degradable starch microspheres for malignant hepatic tumors]. AB - The hepatic artery-embolizing effect of degradable starch microspheres (DSM) was assessed by dynamic CT scanning soon after embolization in patients with malignant hepatic tumors. Using the Seldinger method, DSM with a mixture of contrast medium, MMC, ADM or Epi-ADM was manually injected. The subjects were 32 patients with metastatic carcinoma of the liver (62 treatments) and 15 patients with hepatocellular carcinoma(19 treatments) (47 patients received 81 treatments in all). Dynamic CT scanning was performed within 2 weeks of each embolization procedure, and the percent reduction in the area visualized (necrotic effect) was calculated to assess the efficacy of embolization. The necrotic effect of embolization was classified as CR, PR, NC, and PD after 9, 41, 10, and 2 treatments, respectively, in the patients with metastatic carcinoma of the liver and after 4, 6, 6, and 3 treatments, respectively, in the patients with hepatocellular carcinoma. Although there was no patient in whom the tumor showed 50% or more reduction, the contrast enhanced area showed 50% or more decrease in 60 out of 81 treatments. Therefore, blocking of blood flow seemed to contribute more to the response than enhancement of the efficacy of the anticancer agents. Adverse reactions were all transient and controllable. Based on our results, intra-arterial chemotherapy plus DSM embolization seems to be useful for treating malignant hepatic tumors. PMID- 12557710 TI - [Combined determination of urine uracil levels and plasma 5-FU clearance for a simple order-made treatment with anticancer agents of FU derivative]. AB - Individual differences exist in the pharmacodynamics of fluorouracil-derived anticancer agents, with circadian variability even in the same patient probably due to individual differences in the distribution of dihydrophrimidine dehydrogenase (DPD), a decomposing enzyme. Though DPD activity is usually determined in the liver or blood, a more simplified estimation of DPD activity has been recently attempted using urine uracil levels. However, because urine uracil level has the drawback of being easily affected by food ingestion or kidney function, in this study it was determined simultaneously with the determination of plasma 5-FU clearance after sustained instillation of 250 mg 5 FU, in order to estimate DPD activity more accurately. A correlation was observed between urine uracil levels and 5-FU clearance. In cases showing a baseline urine uracil level below 25.1 mumol/g. Creatinine, the blood concentration decreased due to large 5-FU clearance, with a tendency for diminished efficacy of FU derived anticancer agents. In cases showing a baseline urine uracil level above 99.9 mumol/g. Creatinine, on the other hand, adverse reactions due to FU anticancer agents tended to become more serious. Since urine uracil level can be determined easily, it could be the first choice in screening to detect abnormal metabolism of fluorouracil-derived anticancer agents under present circumstances. By combining determination of urine uracil level with 5-FU clearance, it seems possible to predict adverse effects and the effective rate of these agents more accurately. Under existing circumstances, where genetic analysis remains unavailable as a general practice, the combined determination of urine uracil levels and plasma 5-FU clearance may be beneficial in developing order-made treatments in cancer chemotherapy. PMID- 12557711 TI - [Two patients with distant metastases from head and neck squamous cell carcinomas successfully treated with docetaxel and cisplatin]. AB - Two cases of distant metastases from head and neck squamous cell carcinomas are reported. The patients were treated intravenously with docetaxel (60 mg/m2) in combination with cisplatin (70 mg/m2). The chemotherapy was repeated every three weeks. Both cases showed a remarkable response. A complete response was obtained in one case. Grade 1 nausea and alopecia were observed. One patient had grade 3 neutropenia, which was treated with G-CSF. We herein describe the efficacy of newly developed docetaxel with cisplatin in treating distant metastasis from head and neck cancer. PMID- 12557712 TI - [A case of advanced lateral wall oropharyngeal carcinoma showing complete response to chemotherapy with TS-1]. AB - We report a recent case of squamous cell carcinoma originating in the right tonsil, with metastasis to the right superior cervical lymph nodes (T3N2bM0: stage IVA). Chemo-radiotherapy was administered as first-line therapy; and adjuvant chemotherapy with TS-1 was applied during the subsequent 4-week period for the treatment of the residual tumor in the neck. In this patient (female, 61 years old), the primary tonsillar tumor showed a complete response to the first line therapy, however, the metastatic lesions in the cervical lymph nodes persisted, although the nodes also decreased markedly in size. Subsequent chemotherapy with TS-1 (80 mg/day) given for a 4-week period resulted in complete disappearance of the residual malignancy (CR), as determined by palpation and diagnostic imaging. No serious adverse events occurred during the therapy. Surgical treatment in patients with advanced head and neck cancer often results in a diminished quality of life. Although further accumulation of cases is necessary, TS-1, which allows oral chemotherapy on an outpatient basis, would seem to be a useful drug for adjuvant chemotherapy following radical irradiation in patients with advanced head and neck cancer. PMID- 12557713 TI - [Weekly administration of paclitaxel and pirarubicine for recurrent breast cancer]. AB - The therapeutic efficacy of weekly coadministration of paclitaxel (TXL) and pirarubicin (THP) on docetaxel (TXT)- and epirubicin-resistant recurrent breast cancer, adverse reactions caused by this therapy, and the possibility of ambulatory treatment using it were evaluated. The present study was conducted in 11 patients with recurrent breast cancer with pretreatment with CEF and TXT. The site of recurrence was the lung in 9 patients, lymphnodes in 2, bones in 1, liver in 1 and local foci in 1. One cycle consisted of 20 mg/m2 of THP followed by 80 mg/m2 of TXL 4 h later, repeated three times every other week. Three to six cycles were conducted in each patient. An anti-emetic drug was administered before administration of THP as short premedication. Dexamethasone (16 mg; i.v.) and d-chlorpheniramine maleate (12 mg; p.o.) were administered 1 h before administration of TXL and ranitidine (100 mg; i.v.) was administered 30 min before administration of TXL. Ubidecarenone (30 mg/day; p.o.) was administered for 3 days. The response rate was 27.3% with a rating of PR in 3 patients, NC in 6, and PD in 2. Adverse reactions observed included transient facial hot flushes, alopecia grade 1 or milder grade 1 symptoms, and peripheral nerve damage. No adverse reactions such as myocardial disorders or congestive heart failure were noted. Grade 3 and grade 2 neutropenia occurred in 1 and 6 patients, respectively, and 4 patients were admitted for treatment of this. In conclusion, the short premedication was useful, and this was thought to make it possible to conduct ambulatory treatment with TXL + THP in some patients. The response rate of 27.3%, however, was not satisfactory. It will be necessary to clarify the characteristics of this therapy by administering it to a wider spectrum of patients. PMID- 12557714 TI - [Two cases of effective weekly paclitaxel administration and concurrent radiation for metastatic breast cancer]. AB - We report two cases in which weekly paclitaxel (TXL) administration and concurrent radiation was effective for metastatic breast cancer. TXL (80 mg/m2) was infused over 1 hour after short premedication. Case 1: A 50-year-old woman was found to have atelectasis of the middle lobe after treatment for brain metastasis. She was diagnosed with hilar, mediastinal and supraclavicular lymph nodes metastases. She received weekly TXL administration and concurrent radiation to the mediastinum and supraclavicular fossa. The metastatic lymph nodes had disappeared one month after the treatment. Case 2: A 31-year-old woman was diagnosed with advanced breast cancer with lung, pleural, bone and orbital metastases. She received weekly TXL administration and concurrent radiation to the orbit. The lung and pleural metastases had disappeared and the orbital metastasis was decreased by 75% one month after the treatment, and the case was assessed as a partial response. Leukopenia and other major adverse effects were not observed in either of the two cases. PMID- 12557715 TI - [A case of advanced gastric adenocarcinoma with mild elevation of serum SCC that responded remarkably to adjuvant chemotherapy of ADM, CDDP, ETP and 5-FU (ACVF)]. AB - A 65-year-old man was referred to our hospital because of weight loss. Endoscopic examination and computed tomography (CT) revealed an advanced gastric cancer with multiple abdominal lymph node swellings. Distal partial gastrectomy was performed but lymph node resection was not done, since it was not thought to be curative. Adjuvant chemotherapy was performed for 4 courses with a regimen of ADM 20 mg/m2 day 1, CDDP 50 mg/m2 day 1, ETP 100 mg/day days 3-7, 5-FU 600 mg/m2 every other day on days 3-29. After 3 courses of ACVF therapy, the patient's serum CEA and SCC level normalized and the lymph node metastases became undetectable by CT scan. No severe side effects were observed at any time during the administration of these medications. In this case, serum SCC level was elevated even though histologic examination did not reveal squamous cell carcinoma but poorly differentiated adenocarcinoma. On immunohistochemical analysis, these tissues were stained diffusely with CEA, locally with AE1 + 3, and partially with PAS or Alcian blue. We speculate that this tumor could have developed the potency of SCC secretions without structural change into squamous metaplasia. PMID- 12557716 TI - [A surgically resected case of advanced gastric carcinoma with peritoneal dissemination after treatment with combined chemotherapy of TS-1 and CDDP]. AB - A 60-year-old female had undergone laparoscopic oophorectomy for right ovarian tumor. At the time of surgery, peritoneal dissemination and ascites was observed. Histological examination revealed that the resected ovary, peritoneal nodes and floating cells in the ascites were metastatic adenocarcinomas. Later, the primary malignant lesion was found to be a type 4 gastric carcinoma. The carcinoma was judged to be unresectable and treated by combination chemotherapy with TS-1 and CDDP every 6 weeks. After 3 courses of treatments, upper gastrointestinal series and endoscopic examinations were conducted and revealed a marked reduction of the tumor size. No carcinoma cells were detected by endoscopic biopsy. CT-scan showed complete disappearance of metastatic lesions. Staging laparoscopy was performed for evaluation of the effects of chemotherapy, and no adenocarcinoma cells at peritoneal nodes or ascites were found histologically. We performed total gasterectomy with D1 + alpha lymph node dissection. Histopathologically, resected specimens showed severe fibrosis in most parts of the stomach. Following chemotherapy, the carcinoma was judged to be Grade 2 by histopathological examination. PMID- 12557717 TI - [A case of advanced gastric cancer treated with paclitaxel and TS-1]. AB - We treated a case of advanced gastric cancer with paclitaxel and TS-1. A 64-year old man underwent total gastrectomy, splenectomy, and D2 + No. 16 a 2, b 1 lymph node (LN) dissection for gastric cancer. Computed tomography (CT) revealed metastases of supraclavicular and para-aortic LNs in the 4th postoperative month. Paclitaxel 90 mg was infused once a week, and TS-1 100 mg was administered daily. One course consisted of infusion of paclitaxel for 3 weeks followed by 2 weeks rest and administration of TS-1 for 4 weeks followed by 2 weeks rest. At the end of 4 courses of paclitaxel and 3 courses of TS-1, a partial response of the supraclavicular LN metastasis and a complete response of the para-aortic LN metastasis were achieved. There were no remarkable side effects for 2 years after the operation. This chemotherapy might be suitable to treat patients with LN metastases of advanced gastric cancer. PMID- 12557718 TI - [A case of metastatic gastric cancer responding to weekly administration of paclitaxel as a second-line therapy]. AB - We report a 65-year-old man who underwent distal gastrectomy for advanced and metastatic gastric cancer in June 2000. TS-1 was administered for remnant metastatic lesions as first-line chemotherapy, but a recurrence was found in June 2001. Paclitaxel 80 mg/m2 was then administered weekly in a 1-h infusion on day 1, 8, and 15 as one cycle. After two cycles of paclitaxel administration, the patient was relieved from abdominal pain, and a barium enema revealed marked improvement of the colonic stenosis due to the peritoneal metastasis. There have been few effective chemotherapeutic agents against peritoneal metastasis from gastric cancer to date; however, a weekly paclitaxel regimen is considered to be one of the promising regimens for metastatic and recurrent gastric cancer. PMID- 12557719 TI - [Treatment outcomes with paclitaxel for advanced gastric cancer patients previously treated with TS-1]. AB - In the present report, we describe the treatment results of paclitaxel in patients with metastatic gastric cancer previously treated with TS-1 or combination chemotherapy of TS-1 and CDDP. Paclitaxel was administered to 4 patients at a weekly dose of 80 mg/m2/day for three weeks followed by a one week interval. Remarkable tumor reduction was observed in 2 patients. Case 1: A 52 year-old male patient with gastric cancer and multiple liver metastases was treated by weekly infusion of paclitaxel as a 2nd line chemotherapy. After 1 course, the tumor was remarkably reduced, and the reduction was judged PR. Case 2: A 31-year-old male patient presented with lymphoangitis carcinomatosa and obstructive jaundice resulting from cancerous lymphoadenopathy. After 1 course, chest radiographs and abdominal CT scan showed remarkable reduction of these lesions. The adverse effects observed with this drug were leucocytopenia and liver dysfunction, both of which improved soon. These results indicate paclitaxel is effective for advanced gastric cancer pretreated with TS-1. PMID- 12557721 TI - [A case of metastatic pheochromocytoma with remarkable response to combination of cyclophosphamide, vincristine and dacarbazine]. AB - We report the case of a 58-year-old man with metastatic tumors 13 months after the initial surgery for paraganglioma at the left adrenal gland. A CT scan revealed a large tumor at the right scapula and abdominal paraaortic lymph nodes, and the patient received combination therapy of cyclophosphamide, vincristine and dacarbazine (CVD) every 3 to 4 weeks. After 11 courses, the serum catecholamine level was reduced to the normal range. The metastatic tumors showed optimal reduction in size, and the patient remains alive with no symptoms of the disease one year after the primary chemotherapy. The combination therapy of cyclophosphamide, vincristine and dacarbazine is considered a feasible and effective chemotherapy for metastatic malignant pheochromocytoma. PMID- 12557720 TI - [Two cases of complete response to combination chemotherapy of gemcitabine and docetaxel for recurrent ovarian cancer]. AB - The established standard treatment for advanced ovarian cancer is carboplatin and paclitaxel. However, more than 70% of patients have recurrent disease. The standard therapy for recurrent ovarian cancer has not been confirmed. It was reported that docetaxel had a 30-40% of response rate in patients with recurrent ovarian cancer, and that gemcitabine had a 13-22% response rate. The combination chemotherapy of gemcitabine and docetaxel is also applied to non-small cell lung cancer. We use a regimen of 800 mg/m2 of gemcitabine on day 1 and day 8 in combination with 70 mg/m2 of docetaxel on day 8 with a 3-week interval. We treated 2 patients with recurrent ovarian cancer who responded completely to combination chemotherapy with gemcitabine and docetaxel. PMID- 12557722 TI - [Weekly paclitaxel infusion in patients with recurrent ovarian cancer--a pilot study]. AB - This preliminary study was performed to evaluate the safety and efficacy of weekly paclitaxel administration by 1-hour infusion. A total of 7 patients with recurrent ovarian cancer were treated with weekly paclitaxel (TXL). TXL was given at a dose of 70 mg/m2 in 1-hour infusions every week for at least 20 consecutive weeks unless lesions became progressive. Premedication was administered 30 min before TXL infusion. The 7 patients received a total 110 cycles of therapy. Hypersensitivity reactions were not observed. Grade 3 or 4 neutropenia occurred in only 0.9% of the cycles. Neurotoxicity was commonly observed, but that of grade 3 or greater severity was not recorded. No other severe non-hematologic toxicities were observed. Partial responses were seen in three of 7 patients. Weekly 1-hour TXL is considered to be safe and effective as a salvage therapy in patients with recurrent ovarian cancer. PMID- 12557723 TI - [Bioethics in studies on genetic polymorphisms]. AB - In studies on human genetic polymorphisms, such as single nucleotide polymorphisms (SNPs), little immediate benefit is expected for the individuals contributing the test samples. Such lack of immediate benefits should be noted in the informed consent form obtained prior to sample collection. Ethical considerations are indispensable in obtaining human samples for any type of human gene analysis, and researchers should respect the human rights of the contributors of the samples. Considerable differences in the guidelines for human gene analysis are seen between internationally distributed guidelines and the guidelines of the Japanese government. Lack of education in human genetics in medical schools in Japan could be a reason underlining this difference, and improvement of such situation is needed. PMID- 12557724 TI - Herbal therapy, conventional drugs and quality control. PMID- 12557725 TI - Serious adverse effects between herbal remedies and conventional drugs taken simultaneously. PMID- 12557726 TI - Pre-eclampsia: haemodynamic changes. PMID- 12557727 TI - Pre-eclampsia: multisystem involvement and maternal risks. PMID- 12557728 TI - Occupational and environmental exposure of skin to chemicals. PMID- 12557729 TI - Risks associated with herbal slimming remedies. AB - The market for non-conventional, natural herbal medicines is growing. This marketing opportunity has been seized upon by some unscrupulous practitioners to sell potentially unsafe 'herbal' products. Several people attended a Chinese herbalist for weight loss advice and were sold 'herbal' preparations that contained fenfluramine, a drug which was withdrawn from sale in the West in 1997 after its use was linked to primary pulmonary hypertension and valvular heart disease. Adulteration of Chinese medicines with Western drugs is becoming an increasing problem, and deaths have been reported from Japan and other Far Eastern countries linked to Chinese slimming aids containing N-nitroso fenfluramine. There is a need for increased public awareness of such risks associated with the use of unlicensed medicines; a system of registration for medical herbalists is also required to protect both the reputable practitioners and the general public. PMID- 12557730 TI - Integrating Western medicine and Traditional Chinese medicine in GP surgeries and the community: a review of the two pilot schemes. AB - The popularity of complementary medicine has been relentless over the past decade. Among the most popular are Traditional Chinese Medicine (TCM) which has a long and unbroken history. This article reports on two integrated health care pilot schemes where TCM and Western medicine are merged, to varying degrees, for the benefits of patients. One of the schemes focuses on general medicine whilst the other focuses on mental health. The authors conclude that the integrated schemes have many advantages and, on the whole, are beneficial for patients. The issues involved and learning points are discussed. PMID- 12557731 TI - Strategy development in dealing with violence against employees in the workplace. AB - The creation of health promoting workplaces is a key development in contemporary workplace health. This paper focuses on the issue of workplace violence and the efforts which must be made to address its prevention and to deal with its consequences as a contribution towards developing a health promoting workplace. A critical overview of the issue as it is currently addressed in the international literature is a precursor to the formulation of an inclusive definition of workplace violence. This forms the basis for a framework for the development and implementation of workplace violence policies. The framework takes account of needs assessment, policy formulation in terms of prevention, incident management and reporting, and post-incident support strategies. The needs for effective education and training and good internal and external communications are also explored. The paper concludes by highlighting the necessity for workplaces to take seriously the need for a coherent and well-planned workplace violence strategy. PMID- 12557732 TI - Physicians of the future: Renaissance of polymaths? AB - Science and technology are crucial in modern medicine; societies devote enormous amounts of time, money and effort to developing new diagnostic and therapeutic procedures. However, the fact that people now report higher rates of disability, symptoms and general dissatisfaction with their health and well-being calls us to rethink the functions of health care and medical education. There is a need for a new medical paradigm, which should involve and reconcile the natural and the social scientific paradigms ('two cultures'). Medicine should be viewed as an integrative, biopsychosocial science. Therefore, medical education must involve the study of the biological structures and psychosocial functioning of human beings not as separate systems, but as interactive ones. This mandate suggests that the physician needs to become a sort of 'neo-polymath' in a 'new Renaissance'. The new paradigm, however, should not demand the acquisition of more and more information. Instead, the crucial principle would focus on the appropriate selection of information. PMID- 12557733 TI - Does the stress in childhood and adolescence matter? A psychological perspective. AB - Although stress in adults is well researched and acknowledged, little has been discussed on the existence and reactions of children and adolescents to stress, despite the mounting evidence that they are frequently among the most affected victims of a variety of threatening events. In reality stress is a normal part of a child's or adolescent's life and encompasses common, developmental stressors of daily life and those arising from unusual or traumatic experiences. Apart from more well known stress from family instability, poverty and life transitions, it is easy to forget or ignore the many school pressures and increasingly common social phenomenon of the 'hurried child' in modern life. We can make plans of intervention and prevention only when the condition is recognised by the health care professionals. PMID- 12557734 TI - Reducing waiting times associated with an integrated child health service. AB - Following an increase in average waiting times associated with a child health service in East London, an initiative to rapidly reduce the numbers of children waiting long periods following a referral was undertaken over the period May to June 1999. A multidisciplinary cooperative approach was adopted operating within the existing available resources and involved medical, nursing, managerial and administrative staff. The initiative involved a review of the accuracy of the waiting list, followed by an invitation to remaining patients to provide an option of continuing to wait to be seen or offering attendance at a rapid response clinic associated with reduced waiting and consultation times. Half hourly appointments were routinely offered instead of hourly appointments and proformas were adopted for history taking and onward referrals to save time spent on administration. A total of 162 patients were seen over the course of a month and a satisfaction questionnaire completed by relatives indicated a preference for the new service. The mean waiting time was reduced to under a quarter of the time at the start of the initiative to a mean of less than two months. The purpose of the study was to see if the waiting list could be reduced by using existing staff. We wanted to ascertain the parents' views whether shorter waits and shorter consultation periods were acceptable, and to ascertain if the waiting list could be kept down or whether the waiting list would rapidly recur after the rapid response clinics stopped. The findings are discussed in relation to initiatives elsewhere and the need to maintain a high quality service. PMID- 12557735 TI - Quality of life: an assessment of the state of psychosocial rehabilitation of patients with schizophrenia in the community. AB - The subjective quality of life (QOL) of diabetic patients on oral hypoglycaemics was compared to schizophrenic patients who were well controlled with their antipsychotic medications. This comparison was made using the generic quality of life questionnaire produced by the World Health Organization QOL (WHOQOL) group, namely the WHOQOL-100. Statistical analysis showed that there was no significant difference in the psychological well-being and level of independence between the two groups. However, such measures revealed that the most impaired aspect of well being in the schizophrenic group was the social relationship. Subjective QOL assessment is able to reveal deficits or handicaps that are obscure and probably difficult to appreciate on objective social and clinical evaluation. Such findings are valuable in planning the rehabilitative need of schizophrenic patients in the community. PMID- 12557736 TI - The Essene's sister sect in Egypt: another medical site? AB - The Essenes were, allegedly, the authors of the Dead Sea Scrolls and were settled by the Dead Sea. An ancient source locates their sister sect, The 'Therapeutae', as being by the shores of Lake Mareotis in Egypt. No scholar has previously sought to locate where exactly on Lake Mareotis they were settled. Using clues from Essene sites around the Dead Sea, an attempt is made to suggest a specific location for the Therapeutae. Since there are signs of medical activity at many of the Essene sites, a site with medical association is also sought for the location of the Essene's sister sect in Egypt. A suitable site is found at Canopus. PMID- 12557737 TI - Chinese herbal nephropathy and urothelial malignancy. AB - In Belgium, between 1991 and 1992, seven women under the age of 50 years were admitted to a regional renal dialysis unit with progressive renal failure. All seven had been following the same slimming regimen, prescribed by the same clinic. They had an identical pattern of interstitial fibrosis on renal biopsy. Subsequently it became evident that all were at risk of developing urothelial malignant change. Could there possibly be a link to the prescribed drugs, which included Chinese herbs? PMID- 12557738 TI - Shooting the bearer of 'good news'. PMID- 12557739 TI - Bigger again ... but better? PMID- 12557740 TI - Rightsize--don't downsize. PMID- 12557741 TI - Telemedicine--what's happening now? PMID- 12557742 TI - The link between physician retention and orientation. PMID- 12557743 TI - RVU cost accounting not just for the 'big dogs'. PMID- 12557744 TI - Focus on satisfaction. Using patient focus groups. PMID- 12557745 TI - Make collections your goal. Summit Healthcare case studies. AB - A physician practice management company's approach to improving collections and its application in two different medical groups. PMID- 12557746 TI - Head games. The psychology of medical group financial management and the role of physician leaders. AB - Good practice leaders must understand the psychology of presenting and managing the financial performance of the medical group. Leaders--especially physicians- must strive to understand this psychology to shepherd the group through the various financial challenges it will face. PMID- 12557747 TI - Effective physician governance is not an oxymoron. PMID- 12557748 TI - On with the show. Patient no-shows--some surprising findings. AB - The authors' investigations reveal that the strongest predictor of patient no show rates is insurance status--not waiting time to see a provider. Their findings helped refine scheduling practices to maximize productivity in their organization. PMID- 12557749 TI - Amyloid beta-peptide 25-35 reduces [3H]acetylcholine release in retinal neurons. Involvement of metabolic dysfunction. AB - Cholinergic pathways serve important functions in learning and memory processes. The loss of basal forebrain cholinergic neurons and the presence of senile plaques composed by amyloid beta-peptide (A beta) are found in post-mortem brains of Alzheimer's disease (AD) patients. However, the role of A beta in the cholinergic dysfunction observed in AD is not yet clarified. In this study, we observed that the release of [3H]acetylcholine evoked by K(+)-depolarization was significantly lower in cells treated with A beta 25-35 peptide, than in untreated cells or in cells exposed to the reverse sequence peptide A beta 35-25. The levels of pyruvate, the substrate for pyruvate dehydrogenase, the enzyme involved in acetyl coenzyme A synthesis in the brain, which is rate-limiting for the synthesis of acetylcholine, were significantly decreased, about 40%, in A beta treated cells. A beta 25-35 did not affect choline acetyltransferase activity or [3H]choline uptake. 2-[3H]-deoxyglucose uptake was decreased when cells were exposed to A beta 25-35 or to A beta 1-40. Taken together these data suggest that an impairment of glycolysis, and the consequent decrease in pyruvate levels, may be responsible for the decrement of acetylcholine release observed in A beta treated cells, thus sustaining the hypothesis that the cholinergic dysfunction, observed in AD patients, might be associated with extracellular A beta accumulation. PMID- 12557750 TI - Differential transcription of the mouse acute phase serum amyloid A genes in response to pro-inflammatory cytokines. AB - The acute phase members of the mouse serum amyloid A (Saa) family, Saa1, Saa2 and Saa3, are highly similar at both the nucleotide and protein sequence levels. Saa1 and Saa2 in the BALB/c strain are 72% identical over the first 500 bp upstream of their transcription start sites and to date have been considered to be coordinately regulated. Furthermore, based on their homology with the upstream regions of the human SAA1 and SAA2 genes, it has been assumed that they are Type I acute phase proteins (APPs), i.e. they are primarily regulated by IL-1 and TNF. Here we establish that the BALB/c Saa1, Saa2 and Saa3 genes, in fact, respond differently to IL-1, TNF and IL-6. The Saa1 and Saa2 promoters are strongly induced by IL-6, with synergistic upregulation of Saa2, but not of Saa1, by IL-1 or TNF. In contrast, the Saa3 promoter is strongly induced by IL-1, moderately induced by TNF and only minimally induced by IL-6. We also define important sequence differences between the Saa promoters of Type A (BALB/c and ICR/Swiss) and Type B (129/Ola) strains of mice, that have dramatic qualitative and quantitative consequences for Saa1 and Saa2 regulation. These findings mandate careful strain selection prior to embarking on studies involving mouse models of secondary amyloidosis or cytokine inactivation. PMID- 12557751 TI - Acute-phase, but not constitutive serum amyloid A (SAA) is chemotactic for cultured human aortic smooth muscle cells. AB - The human serum amyloid A (SAA) protein family is subclassified as acute phase SAA (A-SAA), which comprises the SAA1 and SAA2 allelic variants, and constitutive SAA (C-SAA), which is the SAA4 isoform. Extrahepatic production of A-SAA occurs in many organs and tissues of the body, including smooth muscle cells (SMC) of the aorta. A-SAA has been shown to act locally as a chemoattractant for neutrophils, monocytes and lymphocytes via the N-formyl peptide receptor-like (fPRL1). In order to gain further understanding of the physiological significance of local production of A-SAA by SMC, the effect of exogenous A-SAA on the in vitro migration of human aortic SMC was investigated. Increased SMC migration in the presence of A-SAA was detectable after six hours and continued to increase up to 24 hours after incubation. The increased migration was dose-dependent over the concentration range 10 to 100 micrograms/ml. The mode of A-SAA stimulated SMC migration was by chemotaxis not chemokinesis. Exogenous constitutive SAA (C-SAA) did not affect SMC migration. Stimulation of SMC migration by A-SAA was inhibited by both polyclonal and monoclonal antibodies to human SAA1 and also by the inhibitors of fPRL1 signaling, wortmannin, bisindolylmaleimide and pertussis toxin. The results herein indicate that A-SAA, but not C-SAA, may serve as an autocrine factor to influence SMC migration in situations of aortic tissue injury and inflammation. PMID- 12557752 TI - Beta 2 microglobulin serum levels and prediction of survival in AL amyloidosis. AB - To study the relation between beta 2 microglobulin (beta 2M) and survival in AL amyloidosis, we measured the serum level of beta 2M in 80 patients with AL amyloidosis diagnosed within 1 year of evaluation, who had received no therapy. Patients had a median age of 61 years and 52% were male. Major clinical manifestations were renal disease in 25 patients (31%), cardiomyopathy in 23 patients (29%), and neuropathy or other organ involvement in 32 patients (41%). The beta 2M level, measured by an ELISA assay in serum samples collected at the time of evaluation, ranged from 1.69 to 10 mg/ml (mean = 4.57); in 56% of the patients beta 2M > 4 mg/ml. The patients with a beta 2M < or = 4 mg/ml had serum creatinine levels lower than those with beta 2M > 4 (1.43 vs 2.67 mg/dl; p = 0.02). Survival from study entry was analyzed overall by the level of beta 2M, adjusting for creatinine level and clinical stratum. We found the beta 2M level to be predictive of survival (median survival 16.1 months for beta 2M < or = 4 mg/ml vs 8.0 months for beta 2M > 4 mg/ml, p = 0.044). Thus a beta 2M level less than 4 mg/ml indicated a longer time of survival. PMID- 12557753 TI - Presence of variant transthyretin in aqueous humor of a patient with familial amyloidotic polyneuropathy after liver transplantation. AB - To determine the origin of transthyretin (TTR) in the aqueous humor of patients with familial amyloidotic polyneuropathy (FAP), we measured TTR levels and analyzed the TTR forms in the aqueous humor of three FAP patients (one patient; liver transplanted, and two patients; non-transplanted). The total TTR levels were almost the same as reported previously in non-transplanted patients and slightly increased in a transplanted patient. Analyses with mass spectrometry in the two non-transplanted FAP ATTR V30M patients revealed that both wild type and variant TTR forms were detected in their aqueous humor samples. Moreover, variant TTR forms could be detected in the aqueous humor of the transplanted patient while the liver produced no variant TTR. These results suggest that variant TTR in aqueous humor may be derived from retina where TTR was produced. In conclusion, TTR metabolism may occur in its own ocular cycle and variant TTR produced by the retina may play an important role in amyloid formation in the ocular tissues of FAP patients. PMID- 12557754 TI - Senile systemic amyloidosis presenting as bilateral carpal tunnel syndrome. AB - A 74-year-old woman who had developed numbness in both hands was diagnosed as having bilateral carpal and right cubital tunnel syndrome and underwent bilateral carpal and right cubital tunnel release. Transthyretin immunoreactive amyloid deposits were seen on specimens and were also detected in gastric, duodenal and ileal mucosal biopsies. The transthyretin gene analysis showed no mutation. This is a rare case of senile systemic amyloidosis presenting as carpal tunnel syndrome. PMID- 12557755 TI - Serial CT and MRI findings in a patient with isolated angiitis of the central nervous system associated with cerebral amyloid angiopathy. AB - We report serial CT and MRI findings in a biopsy-proven case of cerebral amyloid angiopathy (CAA) with isolated angiitis of the central nervous system (CNS). A 69 year-old man had developed dizziness, dementia, and generalized seizure during the preceding 4 years. An initial examination by brain CT and MRI showed bilateral symmetrical periventricular lesions closely resembling those of Binswanger's disease. Subsequently, the lesions expanded slowly, involving a large area of the right cerebral hemisphere with an obvious mass effect. Since a primary brain tumor was suspected, a brain biopsy was performed, and histopathological examination revealed amyloid beta protein CAA within the meningocortical vessels associated with perivascular monocytic cuffing, indicating the presence of isolated angiitis of the CNS. Multinucleated giant cells containing intracytoplasmic beta protein amyloid around a heavily amyloid laden cortical vessel were also observed. This is the first case report to show sequential radiographical studies of the leukoencephalopathy associated with CAA and isolated angiitis of the CNS. PMID- 12557756 TI - Transthyretin amyloidosis associated with a novel variant (Trp41Leu) presenting with vitreous opacities. AB - We report a 45-year-old woman with a new transthyretin (TTR) variant, substitution of leucine for tryptophan at residue 41, who has showed vitreous opacities without any other visceral organ involvement since age of 42. Congo red staining of vitrectomy specimens revealed that the vitreous fluid contained amyloid fibrils, which were strongly positive for immunohistochemical staining using anti-human TTR antiserum. DNA analysis of the TTR gene showed a G to T transversion at the second nucleotide of codon 41, indicating a replacement of tryptophan (TGG) by leucine (TTG). These results indicate that the patient's vitreous amyloid is associated with this novel TTR mutation. PMID- 12557757 TI - A new transthyretin variant Leu55Gln in a patient with systemic amyloidosis. AB - We report a 40-year-old woman with a new transthyretin (TTR) variant, glutamine replacing leucine at residue 55 (Leu55Gln), who showed progressive somatic and autonomic neuropathy, glaucoma, and vitreous opacities. Symptoms of glaucoma appeared prior to the onset of the neuropathy. Sural nerve biopsy revealed amyloid deposition in the endoneurium, and immunohistochemical analysis was positive with anti-human TTR. Sequencing of the patient's DNA revealed the substitution of A for T at the second position of codon 55 of the TTR gene indicating an amino acid change from leucine to glutamine. PMID- 12557758 TI - Transthyretin mutation (TTRGly47Ala) associated with familial amyloid polyneuropathy in a French family. AB - A French family in which three individuals had familial amyloid polyneuropathy (FAP) was investigated. The proband presented cardiomyopathy with atrial arrhythmia and then developed axonal polyneuropathy, carpal tunnel syndrome, and sclerodactyly. Nucleotide sequencing of exons 2, 3 and 4 of the transthyretin (TTR) gene revealed heterozygosity for a single base change in the second position of codon 47. This G to C transversion predicts replacement of a glycine by an alanine at position 47 in the mature protein. This mutation (G47A) was previously identified in two different families of Italian origin both of which had FAP and cardiomyopathy. Here we report the first identification of this mutation in a non-Italian family. PMID- 12557759 TI - Serum amyloid A; in search of function. PMID- 12557760 TI - [The degradation performance of bioabsorbable acylchitin fiber reinforced PLA composite materials in vitro and in vivo]. AB - The present authors have investigated the degradation performance of acylchitin fiber reinforced polylactide composite materials plates both in vitro and in vivo. The initial flexural strength and the initial flexural modulus of this plate are 114.72 MPa and 3980.05 MPa, respectively. The flexural strength of this plate decreases to 31.42 MPa after the plate has been submerged in injectio natrii lactatic ringeri tissue fluid for a period of 16 weeks at 37 degrees C. Both the in vitro degradation performance and the strength retention of this plate are better than those of the self-reinforced PGA/PLA and PGA, though the initial strength of the latter two being much higher than that of the former. The degradation products of the chitin/PLA composite materials can be absorbed by metabolic pathway. PMID- 12557761 TI - [Study on osteocompatibility of hydroxyl polycalcium sodium phosphate in vitro]. AB - Hydroxyl polycalcium sodium phosphate(HP) is a new substitute material for human hard tissues. The isolated rat osteoblasts from the parietal and frontal bones of 1-3d SpragueDawley rats were used to estimate the osteocompatibility of HP in this study. The effects of the materials on cell morphology, growth and proliferation were determined. The result showed that rat osteoblasts could attach, spread and proliferate on HP as normal cells. The proliferation of cells on HP was more favour able to the adhesion and proliferation of rat osteoblasts and could form osteointegration early. PMID- 12557762 TI - [Studies on the toxicity of polymer of glycolide and lactide(PLGA) and the anti fertility effect of levonorgestrel(LNG)]. AB - PLGA(9:1), a chemical degraded from biomaterials, has been subjected to the tests of acute toxicity, irritations on conjunctiva and muscle, haemolysis in vitro and allergic reaction of mice and rats. The anti-fertility effect of PLGA-LNG microphere composite on the same animals also has been tested. The studies demonstrated that PLGA had fine biological compatibility without side effects such as toxicity, irritations on conjunctiva and muscle, pyrogenicity, haemolysis in vitro and allergic reaction. The contraception period of mice lasted 11.5, 34.6 and 81.2 days respectively after the subcutaneous (s.c.) injection of PLGA LNG microphere which contains LNG 5, 10 and 20 mg/kg separately. The intramuscular(i.m.) injection of PLGA-LNG microphere containing LNG 13.6, 19.8 and 30 mg/kg respectively could keep the estrus of rats supressed for 76.3, 104.1 and 131.8 days. The injections of PLGA-LNG microphere through s.c. or i.m. showed more significant effects than the same dosages of PLGA-LNG micro-crystal. These results suggest that PLGA-LNG microphere injection has long-term anti-fertility effect. PMID- 12557763 TI - [A biomechanical model of local control of microcirculation]. AB - In this paper we analyze the generation and motion of vasoactive metabolites in interstitium, the open and closure of precapillary sphincters, the factors affecting mean arteriolar diameter, and the change of amplitude of arteriolar vasomotion. A biomechanical model of local control of microcirculation has been developed. It is useful in quantitative analysis of microcirculatory local control. PMID- 12557764 TI - [Invasion behavior of hepatocellular carcinoma cells correlates with their rheological properties]. AB - We investigated the viscoelasticity, mechanics of adhesion to collagen I coated surfaces and mechanics of adhesin to liver sinosoidal endothelial cells(LECs) among primary culture human hepatocytes, hepatocellular carcinoma(HCC) cells as well as HCC-Inv cells, namely HCC cells capable of invading the artificial basement membrane, Matrigel. The results showed that the above-mentioned rheological properties of HCC cells differed obviously from those of normal hepatocytes. HCC-Inv cells exhibited higher viscoelastic coefficients, higher adhesion forces to collagen I coated surfaces as well as higher adhesion forces to LECs than those of normal hepatocytes and HCC cells. Immunocytochemistry showed the differences in cell morphology and cytoskeleton structure between HCC cells and HCC-Inv cells. A brief discussion was presented in relation to the changes in cell rheological properties of hepatocellular carcinoma cells and their pathological significance. PMID- 12557766 TI - [A biomechanical study on the retrodiscal tissue of human temporomandibular joint]. AB - In order to investigate the biomechanical properties and functional mechanism of the retrodiscal tissue (RT) of human temporomandibular joint (TMJ), we conducted a uniaxial tensile test on thirteen RT specimens taken from eight fresh TMJs of the human cadavers aged between 8-15 years (m = 11.8 yrs). The experimental data were analyzed in conjunction with the Quasi-Linear Viscoelastic Theory as proposed by Fung YC to characterize the time-dependent behaviour and the constitutive relationship of the RT. The results showed that the physiological biphasic zone of stress-strain curve lied within 20%-30% strain levels. The elastic modulus (MPa), tensile strength(MPa) and strain to failure(%) were high. There were significant differences in strain to yielding and energy resorption (N.mm) between lateral and medial RT. There was a good agreement between theoretical prediction value and experimental result within 6% strain level. These data suggest that there would be a higher passive deformation and lower safety storage of strength in the RT which could be related to anterior displacement of the TMJ disc and that Fung's Theory can adequately describe the stress-time behaviour and the constitutive relationship of the RT within lower strain rate. PMID- 12557765 TI - [Effect of lysophosphatidylcholine on endothelial cell surface expression of adhesion molecules]. AB - Extensive monocyte recruitment is an early phenomenon associated with the development of atherosclerotic lesion, in which adhesion molecules expressed by endothelial cell play an active role. In this study we examined the effects of Lysophosphatidylcholine(Lyso-PC), a major phospholipid component of atherogenic lipoproteins, on the expression of adhesion molecules for monocytes, such as intercellular adhesion molecule-1(I-CAM-1), endothelial-leukocyte adhesion molecule-1(E-selectin) and vascular cell adhesion molecule-1(VCAM-1), in cultured human umbilical vein endothelial cells(HUVECs). After treating HUVECs with different concentration Lyso-PC at different times, the expression of adhesion molecules on the surface of HUVECs induced by Lyso-PC were analyzed using flow cytometry. The results showed that compared with control group, the surface expression of ICAM-1 and VCAM-1 was increased on HUVECs treated with Lyso-PC. ICAM-1 and VCAM-1 expression was maximally induced 1 h after stimulated with 50 micrograms/mg Lyso-PC, compared to basal values. At the 3 h after stimulation, ICAM-1 dropped to basal level while VCAM-1 dropped below to basal level. The effects of Lyso-PC on E-selection expression were negligible with in 3 h. However, at the 3 h of treatment with 50 micrograms/ml, the E-selectin expression on HUVECs began to be up-regulated significantly. We conclude that in addition to its monocyte chemoatractant activity, Lyso-PC can selectively induce ICAM-1 and VCAM-1 expression on the surface of HUVECs. Lyso-PC may play an important role in monocyte recruitment into athersclerotic lesions. PMID- 12557767 TI - [Study on the mechanism of hemocompatibility of titanium oxide films]. AB - In this paper a study on the facts affecting blood compatibility of titanium oxide films synthesized by ion beam assisted deposition is presented, and the mechanism of blood compatibility is discussed. We think that the blood compatibility is determined by surface energy and work function of the film. The protein adsorption is determined by surface energy. An area with a range of values of polar and dispersion components of surface energies, where good protein adsorption property will exhibit, was defined. That is, more human serum albumin (HSA) and less human fibrinogen (HFG) will be adsorbed in this area. Work function is also an important factor to influence blood compatibility. Low work function can inhibit the decomposition of HFG and improve thromboresistance. PMID- 12557768 TI - [An investigation of bioartificial pancreas for treatment of diabetes mellitus]. AB - Bioartificial pancreas(BAP) made up of rat islet, agarose and collagen was developed in this experiment to cure the diabetes mellitus and prevent immune rejection of the islet xenograft. The insulin secretion function, immunoprotection and effect of BAP on the treatment of diabetes mellitus in mice were investigated. The results showed that the BAP maintained the insulin secretion function of islet, decreased the immunogenicity of islet xenograft and normalized the metabolic disturbances of carbohydrate in diabetic mice. These indicate that the BAP composed of the islet, agarose and collagen is an alternate approach to therapy for diabetes mellitus and to remission of immune rejection of the islet xenograft. PMID- 12557769 TI - [Biodynamic response of the human shank subjected to impulse load]. AB - This paper reported the establishment of biodynamic modelling of the human shank in the sagittal palne while the human thigh is fixed. And when the shank is subjected to the two types of externally applied impulse loads, the forces associated with the four main ligaments, as well as the bone-to-bone contact forces in the knee joint are numerically obtained. The contact point locations are also presented together with the angular motions of the lower limb segments. PMID- 12557770 TI - [Design and optimization of a centrifugal pump for CPCR]. AB - Requirements for an optimal centrifugal pump, the vital component in the equipment for cardiopulmonary cerebral resuscitation(CPCR), have been presented. The performance of the Sarns centrifugal pump (Sarns, Inc./3M, Ann arbor, MI, U.S.A) was tested. The preliminarily optimized model for CPCR was designed according to the requirements of CPCR and to the comparison and analysis of several clinically available centrifugal pumps. The preliminary tests using the centrifugal pump made in our laboratory(Type CPCR-I) have confirmed the design and the optimization. PMID- 12557771 TI - [Effect of various assist pumps on recovering the ventricular function]. AB - With the authors' diaphragm pump and pulsatile as well as nonpulastile impeller pumps, and the clinically used Sarns Roller, both left ventricular assist and biventricular assist experiments were performed on mini pigs and calves. The results demonstrate that the pulsatile impeller pump is the best assist pump for recovering the heart function, because it maintains the pulsatility of aortic pressure by heart failure and thus increases the circulation volume, and it enlarges the diastolic pressure so as to increase the perfusion in coronary artery because it has no backflow. PMID- 12557772 TI - [A computer-aided analysis system for counting the synaptic numerical density based on disector technique]. AB - The change in the numbers of synapse is of importance in various kinds of physiological and pathological processes. The numerical density is the structural parameter measuring up the numbers of the particles in the space. A new stereological technique, the disector technique, has been described to count the number of particles in a given volume of tissue. It is a technique for counting the number of arbitrary particles in 3-dimensional space using adjacent parallel sections at known distance apart. In this article also discussed is the software for computer assisted count of the synaptic numerical density based on the physical director technique with Visual Basic 4.0. PMID- 12557773 TI - [Fuzzy space based segmentation algorithm on periosteum medical image processing]. AB - Object description in medical image often has the property of fuzziness, and with the development of computing, fuzzy logical theories are progressively used in medical image processing. Color medical images differ in spacial presentation and fuzzy description from grey-scale images and must be analyzed by special methods. In this paper, a new method of bone cell segmentation based on fuzzy logical theories is presented. With the utilization of fuzzy set theories in the steps of color enhancing, feature extraction and automatical segmentation, bone cells are detected from the background. The method has the advantages of high accuracy and flexibility to many situations. Experiments of bone cell images have proved that it is a fast and effective method. PMID- 12557774 TI - [Electroencephalography inverse problem by subspace decomposition of the fourth order cumulant matrix]. AB - It is an important topic in electroencephalography (EEG) research to localize the EEG activity sources from the scalp recordings. In this paper, based on the fourth-order cumulant matrix, a new sub-space decomposition algorithm is proposed for the EEG inverse problem. As the second-order moments (cumulants) has the drawback of being sensitive to the noise covariance. Using the fourth-order cumulants we need not know the noise covariances, as long as the noise is Gaussian. Computer simulation study on a three-layer concentric sphere head model shows its better performance than the two-order cumulate method in depressing the spatial coherent Gaussian noise. PMID- 12557775 TI - [A noninvasive method for radial pulse-wave velocity and the determinants of pulse-wave velocity]. AB - An effective experimental method has been proposed and used to determine the pulse-wave velocity (PWV) of 257 radial pulse signals. Experimental results show that, compared with the normal pulse, the wiry pulse has an increased PWV(P < 0.01 or P < 0.05), the thready pulse has a decreased PWV(P < 0.05), but the PWV of the smooth pulse is not significantly different from the normal's (P > 0.05). PWV highly correlates with the artery blood pressure. The possible determinants of PWV are analyzed and discussed in reference to clinical diagnosis. PMID- 12557777 TI - [Study of modeling and simulation of the multi-branch cardiovascular system]. AB - By the Power Band Graph(PBG) method, this paper presents a computer simulation model of the multi-branch cardiovascular circulation system, which describes the blood fluid dynamic law in the cardiovacular system by the state equation. The model gives a minute description on physiological characters of cardiovascular circulation system (CVS). An integrate computer model on CVS has been established. The model can simulate physiological characters of cardiovascular circulation system and get the simulation data and the curves of CVS hemodynamics variables. The model can be used widely in the field of physiological system simulation study, medical study, and computer-aided instruction. PMID- 12557776 TI - [Image and quantity analysis of blood-gas in rabbit's artery and Na(+)-K(+) ATPase in their lungs during PE-SWD treated by HFJV]. AB - This study was made to gain an insight into the mechanism of high-frequency jet ventilation(HFJV) treatment for the pulmonary edema after seawater drowning(PE SWD). PaO2, PaCO2 and oxygen saturation (SaO2) of the rabbit and Na(+)-K(+) ATPase in the rabbit's lungs in three groups--PE-SWD group(PE-SWD-G), HFJV group (HFJV-G) and control group(CG), were measured and analysed by the blood-gas analyser and computer image system. The results showed that, after 100 minutes' HFJV, the PaO2, SaO2 and the activity of Na(+)-K(+)-ATPase in the lung capillary endothelial cells in HFJV-G were significantly higher than those in PE-SWD-G(P < 0.01 or P < 0.05). Three parameters of Na(+)-K(+)-TAPase(G1, D1 and D2) in HFJV-G almost returned to their values in CG. The authors suggest that the increase of PaO2 and SaO2 in the rabbit's artery blood in HFJV-G is closely related to the rehabilitation of Na(+)-K(+)-ATPase activity in the lungs after HFJV. The mechanism of successful HFJV treatment for PE-SWD is that HFJV can better correct hypoxemia and improve the rehabilitation of Na(+)-K(+)-ATPase activity in the rabbit's lungs. PMID- 12557778 TI - [A study of the relation between ventricular repolarization duration and autonomic nervous system]. AB - Cardiovascular disease is a growing problem recently. There has been a few attention in the study of ventricular repolarization duration variability. RDV is associated with some cardiac diseases. We studied vagal blockade effects on RDV in urethane-anesthetized rats. To study the relation between ventricular repolarization duration and autonomic nervous system in subjects, we changed the balance of autonomic nervous system by body posture. The HRV and RDV spectral analysis showed that HF peak was modulated by the frequency of controlled respiration. We also found a significant decrease in HFnu and increase in LF/HF in HRV and RDV during vagal blockade. There is a significant increase of LF/HF of subjects in HRV and RDV from supine to tilt, which is consistent with the state of autonomic nervous system. The result of our study is that the ventricular repolarization duration is dependent on the autonomic nervous system. The RDV analysis will be more helpful in the clinic. This approach will provide a more useful noninvasive evaluation of cardiac disease and function. PMID- 12557779 TI - [Study on the mechanism of pulmonary edema after seawater drowning in rabbit]. AB - To study the mechanism of pulmonary edema after seawater drowning (PE-SWD), the indexes of blood-gas and acid-base in rabbits artery blood were measured by the blood-gas analyser. The activity of Na(+)-K(+)-ATPase, cytochrome oxidase(CYTO) and alkaline phospharase(ALP) in the lungs were measured and analysed by computer image system. C-fos mRNA and Fos protein in the lungs were respectively determined by situ hybrioization and immunohisto chemical techniques. The distribution of phospholipid and Ca2+ of rabbits lungs was quantitatively analysed by ultrastructural location method. The results showed that, five parameters of PaO2, oxygen saturation(SaO2), pH, actual bicarbonite(AB) and base excess(BE) and the activity of Na(+)-K(+)-ATPase and CYTO decreased remarkably in PE-SWD. Both c-fos mRNA and Fos protein expression in pulmonary epithelial cells in PE-SWD were significantly elevated compared with the normal controls(P < 0.01). The phospholipids products in the pulmonary alveolar type II epithelial cells were decreased, however, the Ca2+ precipitate pellets inside the lung capillary endothelial cells and the pulmonary alveolar type I and II epithelial cells increased obviously. The arthors suggest that the injuny action of the seawater, hypoxia and metabolic acidosis may be the mian three mechanisms of the pulmonary edema induced seawater drowning. The lowering activity of Na(+)-K(+) ATPase and CYTO in the lungs and calcium overload in the cells are not only evil consequence resulted from above three factors, but also the importent causes leading to the worse of PE-SWD. The transmiting line of Ca(2+)-fos may be also a key link to bring about the worse of PE-SWD. PMID- 12557780 TI - [Noninvasive estimation of tissue temperature in hyperthermia using diagnostic ultrasound]. AB - Hyperthermia is one of the important techniques in tumor treatments. Temperature estimating and monitoring is the key technique in hyperthermia. In this paper, a new nonivasive temperature estimation method based on backscattered ultrasonic power is introduced. Compared with the techniques based on echo-shift and frequency-shift, this method has its advantage for application. The change of backscattered power caused by the temperature change is conspicuous and easy to measure. The new techniques and difficulties in this field are also addressed. PMID- 12557781 TI - [The researches on titanium and titanium alloy in dental use]. AB - Although titanium and titanium alloy have been used in dental field for several years because of their excellent biocompatibility, corrosion resistance and mechanical properties, many practical problems remain to be solved. This review focuses on the practical problems and the prospects of the researches on titanium, including the development of new titanium alloy, the surface treating, the corrosion in oral, and the processability. PMID- 12557782 TI - [Chaos in electroencephalogram]. AB - The history and current situation of EEG study were reviewed in the paper, with an emphasis on the application and the latest developments of chaos in EEG analysis. The problems in present study and the proposals for future investigations were finally discussed. PMID- 12557784 TI - [The mechanism of bone formation promoted by mechano-electrical environments- current studies on local bone factors]. AB - The mechanism for promoting bone formation under the mechanical and the electromagnetical fields stimulation is not yet quite clear. In recent years, it has been found the mechanical and electromagnetical environments may induce the osteogenic cells to produce some local bone factors, such as prostaglandin E2(PGE2), insultine-like growth factors-II (IGF-II), bone morphogenetic protein (BMP) and transforming growth factor beta (TGF-beta). These factors play an important role in bone formation and remodeling. This article introduces current studies on some of these local bone factors under the stimulation of the mechanical and electromagnetical environments. PMID- 12557785 TI - [Artificial skin cultured in vitro]. AB - Artificial skin is a kind of skin which by using tissue engineering techniques, a skin-like material was formed by seeding epithelium cells and dermis fibroblasts on a good biocompatible material after proliferation. The trauma restoration and reconstruction were achieved by transplanting the skin graft an skin defects after the artificial skin graft cultured in vitro. Artificial skin research is a new active field in tissue engineering. It has wonderful prospect for clinics to radically settle the problem of restoration of skin defects. The recent studies on the models, cultured methods and influenced factors for epithium culture, dermis culture and artificial skin in vitro were reviewed in this paper. PMID- 12557783 TI - [Reconstruction technology of electrical impedance tomography]. AB - Electrical impedance tomography(EIT) is a new medical image technology; it has the advantages of low cost, nonionizing radiation, and the capability of production images of physiological function dynamically. EIT reconstruction has always been a project of importance to EIT researchers. In this paper, a review of the development of EIT reconstruction is presented. PMID- 12557786 TI - [The research progress of using electromagnetic technology in treatment of bone diseases]. AB - In these years, the study of the bioeffects of electromagnetic fields (EMFs) has been listed as the national important task in the field of biological physics. Using EMFs to treat bone diseases is associated with electric technology, biology and medicine. Previous medical studies have shown that EMFs can stimulate healing in bone fractures. But the effects are still controversial and the mechanism is not yet clear. Therefore, it is necessary to carry out experiments in vitro at cellular and molecular levels. Research workers have been exploring the effects of EMFs on bone formation and metabolism. The suggestion of using EMFs in the treatment of osteoporosis and other osseous diseases has been put forward. PMID- 12557787 TI - [An experiment on disinfection using high power microwave]. AB - This experimental disinfection was intended to examine the effect of high power microwave on bacteria, and the putative nonthermal effect. The bacteria of the experimental groups were irradiated by high power microwave in different intensities and time and were compared with the controls--bacteria heated only. The principles of disinfection by means of high power microwave were studied. The results showed that the temperature did not change after the dry bacteria were irradiated by high power microwave in different intensities and time, but the number of bacteria alive decreased significantly(P < 0.05). The high power microwave's effect of disinfection was greater in the wet bacteria group than in the dry bacteria group and the bacteria heated group. These data indicate that the disinfection effect of high power microwave is higher than that of heating. High power microwave can kill dry bacteria, and it has the nonthermal effect in addition to its thermal effect. PMID- 12557788 TI - [A study on the determination method of biological function of BRF]. AB - This study was aimed to establish a method for determining the biological function of biological wave regulating factor (BRF). Proteus mirabilis was used to set up the experimental model. The biological function of the component obtained from wave growth media beneath the bacterial colony rings resulting from bacterial proliferation was determined in biological wave test by analytically microbiological method. The results indicated that BRF could enhance biological wave and shorten its period. The method is of some value in studying the regulative mechanism of biological wave. PMID- 12557789 TI - [Studies of adsorbents for hemoperfusion--preparation and in vitro studies of cross-linked agar beads entrapped Attapulgite clay]. AB - A new method for preparation of cross-linked agar beads entrapped Attapulgite clay (CAA) is reported. Attapulgite clay was coated with agar and shaped in organic solvent, as well as cross-linked by epichlorohydrin. The products withstood autoclaving at 121 degrees C for 30 minutes and had a good adsorption ability for model compounds and medicines. The results of preliminary investigation indicate that CAA is relatively hemocompatible. PMID- 12557790 TI - [Alteration of free radical generation in pulmonary tissue after quick decompression]. AB - The purpose of this experiment was to evaluate decompression-induced pulmonary injury by means of measurement of ascorbate free radical (Vc-.) alteration in pulmonary tissue of the rats which experienced rapid decompression. 29 male S-D rats were divided into 4 groups: A was executed at 45 min post-decompression; B was control for A; C was executed at 90 min; D was ontrol for C. Group A and group C were to compress up to 0.5 MPa(air) and stay for 60 min, then were decompressed smoothly to normbaric in 1 min. The 0.05 M N-tert-butyl-alpha phenylnitrone (PBN) was given at 20 min after completion of decompression (groups A and B) or at 65 min (groups C and D). At 25 min after PBN injection the animals were killed by cutting arteria coeliaca, and 0.75 g pulmonary tissues were taken. These tissues were homogenized for electron paramagnetic resonance (EPR) assay. The results demonstrated that the ascorbate free radical (Vc-.) signals were obtained at all samples. The intensity of signals measured in group C rose markedly (P < 0.05) and that in group A was close to control's. So it can be concluded that rapid decompression would increase the generation of free radical in lung. PMID- 12557791 TI - [Zero-stress state of renal artery in spontaneously hypertensive rats]. AB - By observing the opening angle of the renal artery, the zero-stress state of the renal artery was studied in spontaneously hypertensive rats (SHR) prior to and posterior to their hypertension, and the role of endogenous angiotensin II (Ang II) and endothelin (ET) on the zero-stress state was studied by giving SHR Losartan(AT1 receptor antagonist of Ang II) and BMS-182874 (an A type receptor antagonist of ET) respectively. The opening angle of the renal artery in SHR with established hypertension was much greater than that in wistar-Kyoto rats(WKY) (114.2 +/- 42.2 vs 70.2 +/- 30.4, P < 0.01). The opening angles of the renal artery in SHR treated with Losartan and BMS-182874 were much smaller than those in control SHR. The opening angles of the renal artery in Losartan-treated and BMS-182874-treated SHR were 65.9 +/- 32.7 and 66.6 +/- 41.54 respectively. The results suggest that non-unequal growth exists in the renal arterial wall in SHR with established hypertension and the residual stress and strain in SHR are greater than those in age-matched WKY. Endogenous Ang II and ET may play a role in the changes of zero-stress state in SHR with hypertension. PMID- 12557792 TI - [A practical compression algorithm of the X ray images]. AB - The application of a lossless compressioin algorithm, based on the R, G, B value of pixel, to a X ray analyzing system is introduced through some medical examples of the X Ray image. The realization of the algorithm is completely presented, based on the design and the description of the characteristics, the R, G, B value of the pixed and the lossless compression methods of the X ray images. PMID- 12557793 TI - [The development of electronic cold and heart therapeutical instrument]. AB - This paper introduces a kind of the rapeutical instrument with ecletctronic cold and heat. It make use of the technology of semiconductor refrigeratio. It have many probe. It apply to different focus treatment. The instrument have achieved cold and heat treatment and accurate temperature mesure and control for human body tissue trauma. PMID- 12557794 TI - [Vaccination against influenza in the aged and high-risk persons]. PMID- 12557795 TI - [Studies on cardiovascular prevention focusing on nutritional interventions: quantitative and qualitative research]. AB - Dietetic education is today considered one of the key elements of preventive and therapeutic strategies in cardiovascular medicine. The combination of quantitative and qualitative research permits the linking of the effectiveness and efficiency of the interventions targeted to cardiovascular prevention, placing at the centre of the clinical scenario the individual subject and giving due attention to his/her global health status and quality of life. PMID- 12557796 TI - [Nutritional status and bone mineral mass in children treated with ketogenic diet]. AB - Ketogenic diet (KD) is a high fat (90%), low carbohydrate (3%) diet used to treat refractory seizures in child. This highly unbalance diet could damage nutritional status. The aim of this study is to evaluate if KD can affect on growth and on mineral status in child. Seven child (1 females and 6 males) age between 3-16 years were retrospectively studied to assess nutritional status during KD; we evaluated anthropometric measurements (weight, height, skinfold and circumferences), bone mineral content and bone mineral density, using x-ray energy absorptiometry (DXA) and some biochemical parameters. We have not found any short term modifications (six months) concerning growth, and biochemical parameters studied. KD could worsen bone mineral status. PMID- 12557797 TI - [Cut-off level of glycosylated hemoglobin indicating the development of complications in type 2 diabetes mellitus]. AB - Prevention of chronic complications is very important in the management of type 2 diabetes, and the availability of a laboratory test individuating at risk subjects should have a marked usefulness. This study sought by the method of ROC curves the cut-off of glycated haemoglobin discriminating the subjects with microvascular, neurologic and renal complications from those without. The cut-off was identified at a glycated haemoglobin level of 7.95, showing a sensitivity of 74.8% and a specificity of 73.3%, which makes it a possible tool for evaluating the risk of complications in type 2 diabetes. PMID- 12557798 TI - [Nasopharyngeal tuberculosis]. AB - Nasopharyngeal tuberculosis is a rare condition and, sometimes, associated with a cervical lymphadenopathy. The authors report a case of an isolated nasopharyngeal tuberculosis in a 69-year-old female with otological symptoms. The case is remarkable because of its coming in a non-endemic zone for the tuberculosis, and because of its probable persistence by many years. PMID- 12557799 TI - [Clinical surveillance of patients with chronic recurrent pancreatitis]. AB - Clinical surveillance of patients affected by chronic relapsing pancreatitis is not a particularly difficult task. In fact, a number of precise landmarks is available such as recurrence of pain and nutritional conditions, possibly enriched by biochemical (specially serum pancreatic enzymes) and imaging data (US, CT, MR), which also may be periodically scheduled. Despite the opinion of some experts, the best timing for the periodical checks should probably be decided on an individual basis. A suitable surveillance program may also be useful to favour the patient compliance and to implement any therapeutic measures. PMID- 12557800 TI - [Nocturnal desaturation in chronic obstructive pulmonary disease]. AB - Chronic obstructive pulmonary (COPD) patients with mild-moderate daytime hypoxia or normoxia (PaO2 > 60 mmHg) may also have transient oxygen desaturation during sleep. Revisiting literature, the authors consider various features of this condition: the prevalence, causes, diagnostic procedures, symptoms, predisposing factors, consequences and therapy. The conclusion is that isolated nocturnal oxygen desaturation in COPD patients is a pathologic condition not yet sufficiently cleared; it likely represents an intermediate step in the evolution of COPD to overt respiratory failure. Since negative outcomes are possible, nocturnal oxygen desaturation must be early put in evidence and then treated with suitable drugs, correct dietetic regimen, respiratory physiotherapy and nocturnal oxygen-therapy. PMID- 12557801 TI - [Misoprostol and oxytocin for induction of cervical ripening and labor in patients with term pregnancy and premature membrane rupture]. AB - OBJECTIVE: To evaluate the effect of the concurrent administration of intravaginal misoprostol and oxytocin for cervical ripening and labor induction on length labor, mode of delivery and perinatal outcomes. PATIENTS AND METHODS: One hundred seven patients with singleton pregnancy at term, vertex presentations, premature rupture of membranes and Bishop scores of < or = 4 were randomly assigned to receive one of three treatments: Group I: Intravenous oxytocin plus intravaginal misoprostol (n = 36); Group II: Intravenous oxytocin plus placebo intravaginal (n = 34); Group III: Intravaginal misoprostol plus intravenous placebo. The time interval from induction to beginning of the labor, from induction to delivery, mode of delivery and perinatal outcomes were measured. RESULTS: The mean time from induction to beginning of labor was different between the groups: Group I: 48.75 minutes, Group II: 107.50 minutes, Group III: 95.94 minutes (p = 0.0024). The mean time in minutes from induction to delivery was different between the groups: Group I: 359.83; Group II: 537.05; Group III: 474.54 (p < 0.05). The frequency of tachysystole, mode of delivery and perinatal outcomes were similar among the three groups. CONCLUSIONS: Oxytocin that is administered simultaneously with intravaginal misoprostol for cervical ripening and labor induction in patients with pregnancies at term, premature rupture of membranes and Bishop scores < 4 make the labor beginning quickly, significantly shortens induction to delivery times without affecting the mode of delivery and with no apparent adverse maternal and perinatal effects. PMID- 12557802 TI - [Sex "addiction": compulsion and controversy]. AB - Modern occidental society often distorts and, in some cases, ignores healthy concepts regarding sexuality. Besides, It not always considers this, to be mental health or to represent an expression of whole health. Such ambivalence towards sex and the sexual is located between the limits of the sacred and erotic, both associated to taboos and their transgression. Since the last century, the current "neosexual" revolution intents to dismantle old patterns in favor of the dissociation of human sexual sphere, the dispersion of sexual fragments towards individualism and intimate relationships diversification. Within such context, "addiction" to sex and the sexual, as a compulsive conduct, represents on one side a clinical reality each time better observed and diagnosed, where the importance of family history is recognized, as well as individual psychopathology, and marital life, in its etiology and also for its treatment and prevention. On the other hand, it has not been classified in the mental health catalogues as it is not considered, meanwhile, as the problem is been maintained, the co-morbidity and its consequences multiply, and the controversy continues. PMID- 12557804 TI - [Acardic and acephalic fetus in the sequence of disruption of inter-twin transfusion syndrome: report of a case with autopsy]. AB - An acardic, acephalic monster is a rare condition occurring once in every 35,000 deliveries. This report details one case occurring in a twin pregnancy in a 19 year old primigravid patient who has spontaneously aborted at 21 weeks gestation. Both twins were males, one of the them weighted 280 g and was acephalic. Autopsy study showed to be also acardic with rudiments of the gastrointestinal tract. The other weighted 350 g and was normal. The placenta was monoamniotic/monochorionic and both umbilical cords had three vessels on gross examination. We think that the scanty literature in Mexico is due to reluctance to report isolated cases like the one we comment here, and depending on local interest in teratology. The salient feature of the pertinent literature, as well as some clinical, radiological and pathogenic considerations are made. PMID- 12557803 TI - [Hormone replacement therapy with transdermal estradiol lowers insulin-cortisol and lipoproteins levels in postmenopausal women]. AB - Increased levels of circulating insulin and cortisol, interpreted as part of aging process, have been associated with an increase risk of developing cardiovascular disease and diabetes mellitus. Because estrogens affect insulin balance, hypoestrogenism in menopausal women may lead to elevations in both insulin and cortisol. The purpose of the present study was to determine the effect of transdermal estradiol administration on the insulin-cortisol binomial. A prospective study was carried out in 30 menopausal women aged 48 to 55 yr, receiving transdermal estradiol 50 micrograms/day during three months. Ten healthy menopausal women (49 to 58 yr) were the control group. Serum levels of cortisol, insulin, lipoproteins, and leptin were quantified by specific assays before and after 3 months of transdermal estradiol therapy. Baseline cortisol levels decreased significantly from 143.4 +/- 10.6 ng/mL to 110.2 +/- 6.7 ng/mL (M +/- SE) (p < 0.001) after 3 months of transdermal estradiol. In parallel, augmented baseline insulin levels diminished significantly from 26.1 +/- 2.0 microlitersU/mL to 21.7 +/- 1.2; (M +/- SE) (p < 0.05). Glucose level were unaffected by this therapy, but it was restored to normal the augmented baseline levels of both triglycerides and low-density cholesterol. Total cholesterol and high-density-cholesterol as well as circulating leptin were unchanged. Transdermal estrogen induced-decrease in circulating cortisol, insulin, triglycerides, and low-density cholesterol to normal values may have a beneficial metabolic effect in menopausal women. PMID- 12557805 TI - [Maternal mortality. A challenge of the new millennium]. AB - Maternal mortality is a principal concern of the contemporary society because of consequence to generate since the most cases to be about young woman and healthy, whose death be able to avoid. This study analyzed the cases registered in Aguascalientes delegation of IMSS during ten years with purpose to find strategy for to salve. The case fatality rate was average of 26.1 x 100,000 alive births. The principal causes was the obstetric hemorrhages; preeclampsia and lung thrombosis. The greater percentage was in woman aged between 25-39 years and multiparous. Most of 50% doing appropriate prenatal control. A greater percentage was pregnancy of term with 77.4 a live births and 59% performed cesarean sections. 93% was through the puerperium. Only 20% was to classify as treatment inappropriate and inopportune and 74% was maternal mortality foreseeable. Some type responsibility existed in 38.7%. PMID- 12557806 TI - [Perinatal mortality: a proposed analysis]. AB - Perinatal mortality is an epidemiologic indicator that evaluates materno-infantil quality care indirectly. Obtaining rates of perinatal mortality from continuous and confident information systems allow us a more precise evaluation of quality care in developed countries. In developing countries as Mexico, there aren't yet evaluating health programs that let us to know the actual situation of our medical care. Being the main mistake subregister problems in regional and national perinatal mortality case. All this let us to a not optimum use of the limited resources given for health. The aim of this paper is to propose an analysis way into this matter for knowing our institutional and national situation. This would permit us to suggest effective strategies in the prevention of obstetric complications as in the preterm birth case. The main objective is to prevent death in the perinatal period. PMID- 12557807 TI - [Clinical and demographic profile of patients with gyneco-obstetric disease colonized by Streptococcus agalactiae]. AB - OBJECTIVE: To describe demographics and clinical features in women with vaginal colonization with Streptococcus agalactiae attended in a gyneco-obstetric hospital. METHODS: Descriptive study of 118 women with isolation of Streptococcus agalactiae in cervical swab during January 1992 to December 1998. Major patient features were obtained in retrospective revising their chart and were classified in demographic, gyneco-obstetrical and clinical characteristics. Statistical analyses were performed with descriptive statistics using central tendency measurements and dispersion, frequencies, rates and proportions with all different variables. RESULTS: Mean age was 31 +/- 11 years; 44 (37.3%) received attention because of sterility, 37 (31.4%) were pregnant, 26 (22%) had any other gynecological disease and, 11 (9.3%) were in a group of high pregestational risk. One hundred eleven (94.1%) were sexually active, starting at a mean age of 20.9 +/- 4.3. GBS was the only microorganism isolated in 70 patients (59.3%). A concomitant cervicovaginal infection was identified in 22 (18.6%) patients. A sexual transmitted disease was present in 25 (21.2%) patients, with isolation of human papilomavirus (HPV), M. hominis, C. trachomatis y T. vaginalis. Fifty-five patients had a concomitant illness: 24 with endocrine disease, 12 cardiovascular pathology and 10 with oncologic disease. CONCLUSIONS: Vaginal colonization of mexican women by GBS is low. However, clinical and demographics characteristics are required to identify these women easily, because of the importance of maternal and infant infection by this organism. PMID- 12557808 TI - [Extra-uterine term pregnancy]. PMID- 12557809 TI - Women's health and gender biology: the late but welcome arrival of evidence-based research. PMID- 12557810 TI - Soy foods and soybean isoflavones and menopausal health. AB - Soybeans are a natural dietary source of isoflavones, which have estrogen-like properties. Therefore, it is worthwhile to consider the implications for soy of the recently published findings of the Heart and Estrogen/Progestin Replacement Study (HERS) I/II and the Women's Health Initiative (WHI). The WHI found coronary heart disease (CHD) risk to be increased in women receiving hormone replacement therapy, and both studies found increases in venous thromboembolic disease in such women. Additionally, stroke and breast cancer risk were increased in the WHI, although risk of colorectal cancer and fracture was decreased. Because research suggests that it is the combination of estrogen plus progestin, and not estrogen alone, that increases breast cancer risk, soy seems unlikely to increase risk because it has no progestin activity. Similarly, there is no evidence to suggest that soy will increase venous thromboembolic disease or stroke; however, only limited data are available in this area. There are promising data suggesting that soy may decrease CHD risk, although studies conducted thus far have examined only markers of risk and not actual CHD events. Similarly, short-term studies generally suggest that soy reduces bone loss in postmenopausal women; however, such effects have been noted primarily only at the spine, and longer-term studies are needed. Finally, very limited human research suggests that soy may decrease colon cancer risk, but this is highly speculative. The results of HERS I/II and WHI suggest that soy may have some of the advantages, but not the disadvantages, of combined hormone replacement therapy (at least with respect to the specific hormones and doses used in the HERS I/II and WHI), but that large, long-term intervention studies examining disease outcome are needed before definitive conclusions can be drawn. Nevertheless, the evidence warrants recommendations that menopausal women include soy in their diets. PMID- 12557811 TI - Black cohosh: an alternative therapy for menopause? AB - Due to the long-term health risks now associated with hormone replacement therapy, many menopausal women are actively seeking alternative treatments. One such alternative is black cohosh (Actaea racemosa, syn. Cimicifuga racemosa), which has been used in the United States for the treatment of gynecologic complaints for more than 100 years. Review of the published clinical data suggests that black cohosh may be useful for the treatment of menopausal symptoms, such as hot flashes, profuse sweating, insomnia, and anxiety. Results from the most recently published trial, however, indicate that black cohosh is not effective for the treatment of menopausal symptoms in breast cancer survivors being treated with tamoxifen. Because the overall quality of the published clinical trials is low, two new randomized, double-blind, placebo-controlled clinical trials are currently underway in the United States. To date, only one standardized black cohosh extract has been tested clinically; the current recommended dose is 40-80 mg per day. At least 4-12 weeks of treatment may be required before any therapeutic benefits may be apparent. Adverse reactions such as nausea, vomiting, headaches, dizziness, mastalgia, and weight gain have been observed in clinical trials. No drug interactions are reported in the medical literature. The estrogenic effects of black cohosh are controversial, and the more recent data indicate that black cohosh extracts may have an anti-estrogenic activity. Owing to potential effects on sex hormones, however, black cohosh should not be administered to children or during pregnancy and lactation. PMID- 12557812 TI - The obese patient with infertility: a practical approach to diagnosis and treatment. AB - Overweight and obesity are serious and prevalent conditions in Western countries and carry many health consequences, including reproductive dysfunction. In particular, excess fat in the abdominal area is strongly related to disorders of the reproductive system. Moderate weight loss and reduction of abdominal fat improves menstrual regularity, ovulation, and infertility in women. This may be etiologically related to insulin resistance, particularly in a subset of infertile women with polycystic ovary syndrome. As such, weight loss should be promoted as an initial treatment option for obese women with infertility. However, the most effective method for achieving and maintaining weight loss is unclear. Gradual weight loss is best achieved through a sensible eating plan that can be maintained over long periods of time. The likelihood of maintaining weight loss is increased when diet is combined with regular exercise, cognitive behavior therapy, and a supportive group environment. Adoption of these principles in a primary healthcare setting can therefore aid in treatment of infertility related to obesity. PMID- 12557813 TI - Evaluation and treatment of low bone density in anorexia nervosa. AB - The prevalence of anorexia nervosa has increased in recent years, and a large proportion of adults as well as adolescents with this disorder have low bone density and, therefore, an increased risk of fractures. Anorexia nervosa often begins during adolescence, when peak bone mass is accumulated, resulting in significant deficits in bone mass accrual. Therefore, it is important to determine bone mineral density in adolescent and adult women who have this disorder, and to improve or at least stabilize bone metabolism in those with low bone mass. To do this, it is necessary to understand the mechanisms underlying low bone density in anorexia nervosa. This article discusses current concepts related to bone loss associated with anorexia nervosa, including how to prevent it. PMID- 12557815 TI - Violence against psychiatric nurses: team approach is needed. PMID- 12557816 TI - PRN medication use in inpatient psychiatry. AB - 1. Administration of p.r.n. medications occurs frequently for staff nurses in inpatient psychiatry. 2. The appropriate use of p.r.n. medications is not well addressed in nursing textbooks or hospital policies and procedures. 3. Thorough assessment of patients is an essential step in the safe administration of p.r.n. medications. 4. Concise, accurate clinical evaluations may be the most successful way to determine the best dose of medication for each patient. PMID- 12557818 TI - Child and adolescent homicide survivors. Complicated grief and altered worldviews. AB - The act of homicide may influence the worldviews of children and adolescents. Problematic beliefs of uncertainty, inadequacy, perceiving the world as dangerous, self-denial, and lack of control can contribute to complicated grief in children and adolescents, and can potentially disrupt their normal psychosocial growth and development. Mental health professionals' understanding of grief after the homicide of a family member enhances their ability to intervene with and support young people struggling to cope with and adapt to a sudden loss. PMID- 12557817 TI - The Four S Model in action for de-escalation. An innovative state hospital university collaborative endeavor. AB - In this article, the Four S Model is presented as a working clinical framework for generating structured nursing interventions and treatment plans. The de escalation process, formalized by the De-escalation/Alternative to Restraint Flowsheet, is one way in which the model is being operationalized to reduce seclusion and restraints. As shown by the flowsheet, the model allows for integration of mental health and mental illness. It spans the scope of nursing practice at all levels, from para-professional to advanced practice nurse, by integrating the mental health concepts of communication, leadership, clinical judgement, critical thinking, and caring within its framework. Most important, the flexibility to individualize treatment that is built into the model allows for generalization to all psychiatric presentations. PMID- 12557819 TI - Defining healthy dating in an adolescent psychoeducational group. PMID- 12557820 TI - What's the difference between these modifiers? PMID- 12557821 TI - Five ways to save money on printers. PMID- 12557822 TI - Can your patient's victim sue you? PMID- 12557823 TI - What an FP's week is really like. PMID- 12557824 TI - Compliance: focus on the basics. PMID- 12557825 TI - When you ask for payment, do it right! PMID- 12557826 TI - Bone scans at the drugstore? PMID- 12557827 TI - Where has all the money gone? PMID- 12557828 TI - It's med school, not boot camp. PMID- 12557830 TI - Frank words from the top. Interview by Wayne J. Guglielmo. PMID- 12557829 TI - Medicare cuts. Will patients pay the price? PMID- 12557831 TI - Legal risks of telemedicine. PMID- 12557832 TI - [Patient with increased stomach cancer risk. Helicobacter can cause damage here]. PMID- 12557833 TI - [Patients with osteoporosis need calcium and vitamin D3. Despite dietary recommendation often malnutrition]. PMID- 12557834 TI - Tracheal intubation without muscle relaxants: remifentanil or alfentanil in combination with propofol. AB - BACKGROUND AND OBJECTIVE: In some situations, the use of muscle relaxants (neuromuscular blocking drugs) are undesirable or contraindicated. We compared intubating conditions without muscle relaxants in premedicated patients receiving either alfentanil 40 microg kg(-1) or remifentanil 2, 3 or 4 microg kg(-1) followed by propofol 2 mg kg(-1). METHODS: In a randomized, double-blind study, 80 healthy patients were assigned to one of four groups (n = 20). After intravenous atropine, alfentanil 40 microg kg(-1) or remifentanil 2, 3 or 4 microg kg(-1) were injected over 90 s followed by propofol 2 mg kg(-1). Ninety seconds after administration of the propofol, laryngoscopy and tracheal intubation were attempted. Intubating conditions were assessed as excellent, good or poor on the basis of ease of lung ventilation, jaw relaxation, laryngoscopy, position of the vocal cords, and patient response to intubation and slow inflation of the endotracheal tube cuff. RESULTS: Seven patients who received remifentanil 2 microg kg(-1) and one patient who received remifentanil 3 microg kg(-1) could not be intubated at the first attempts. Excellent intubating conditions (jaw relaxed, vocal cords open and no movement in response to tracheal intubation and cuff inflation) were observed in those who received either alfentanil 40 microg kg(-1) (45% of patients) or remifentanil in doses of 2 microg kg(-1) (20%), 3 microg kg(-1) (75%) or 4 microg kg(-1) (95%). Overall, intubating conditions were significantly better (P < 0.05), and the number of patients showing excellent conditions were significantly higher (P < 0.05) in patients who received remifentanil 4 microg kg(-1) compared with those who received alfentanil 40 microg kg(-1) or remifentanil 2 microg kg(-1). No patient needed treatment for hypotension or bradycardia. CONCLUSIONS: Remifentanil 4 microg kg(-1) and propofol 2 mg kg(-1) administered in sequence intravenously provided good or excellent conditions for tracheal intubation in all patients without the use of muscle relaxants. PMID- 12557835 TI - Lack of prediction of mefloquine and mefloquine-artesunate treatment outcome by mutations in the Plasmodium falciparum multidrug resistance 1 (pfmdr1) gene for P. falciparum malaria in Peru. AB - We assessed whether mutations in the Plasmodium falciparum multidrug-resistance gene 1 (pfmdr1) (C1034S, D1042N, and Y1246D) would predict treatment outcome during a 28-day in vivo treatment trial in the Peruvian Amazon. Mefloquine (MQ) was compared with mefloquine-artesunate (MQ-AS) in a randomized, multi-clinic protocol for the first time in the Americas. Of 115 patients enrolled in the in vivo arm, 97 patients were eligible for molecular analysis. All 97 patients remained parasite-free during 28 days of follow-up (MQ, n = 46; MQ-AS, n = 51), indicating 100% clinical efficacy of the MQ and MQ-AS treatment regimens. The reported MQ-sensitive alleles (C1034, D1042, and Y1246) were present in 48.5% (n = 47) of the cases, whereas 49 isolates (50.5%) contained the D1246 mutation reported to confer MQ resistance in vitro. However, neither this mutation nor a double mutation (S1034, D1246; n = 16) was predictive of MQ treatment outcome. PMID- 12557837 TI - Does the International Consensus Statement on ADHD leave room for healthy scepticism? PMID- 12557836 TI - Efficacy of chloroquine, sulfadoxine-pyrimethamine, and mefloquine for the treatment of uncomplicated Plasmodium falciparum malaria on the north coast of Peru. AB - As part of an effort to assess antimalarial drug resistance in Peru, we carried out 14-day in vivo efficacy trials of chloroquine (CQ; 25 mg/kg) and sulfadoxine pyrimethamine (SP; 25 mg/kg of the sulfadoxine component) for the treatment of uncomplicated Plasmodium falciparum infections at three sites on the northern coast of Peru. Mefloquine (MQ; 15 mg/kg) also was evaluated at one site. The results from all three sites were similar. Of the 53 patients treated with CQ, 58.5% had RII/RIII responses. No RIII failures were observed among the 112 patients who received SP, but 4.5% and 1.8%, respectively, had RII and RI responses. All 33 patients treated with MQ showed a sensitive response. Early treatment failures were observed in 27.1% of the CQ patients but in no patients receiving SP or MQ. Late treatment failures were seen in 59.3% of the CQ patients and 6.4% of the SP patients but in none of those treated with MQ. Based on these findings and because of concern about the potential for development of resistance if SP were used alone, the National Malaria Control Program is planning a change in malaria treatment policy to SP-artesunate combination therapy for this region of the country. PMID- 12557838 TI - Towards a better drug treatment for patients in child and adolescent psychiatry. The European approach. PMID- 12557840 TI - Abstracts of the 32nd Annual Meeting of the German Society for Thoracic and Cardiovascular Surgery. February 23-26, 2003. Leipzig, Germany. PMID- 12557842 TI - [German Society of Psychiatry, Psychotherapy and Neurology 2002 Congress. Berlin, Germany, 27-30 November 2002. Abstracts]. PMID- 12557841 TI - [Historical evolution of cochlear implants--from Volta to multichannel intracochlear stimulation]. PMID- 12557843 TI - [Surgical techniques: Plastic reconstructive surgery of the face. Cheek area. 1999]. PMID- 12557844 TI - [Which studies are necessary in the hemophilic patient before surgery? What precautions are necessary?]. PMID- 12557845 TI - Abstracts of the 26th Congress of the German Society of Hypertension. 13-16 November 2002. PMID- 12557846 TI - Rugby union. PMID- 12557847 TI - [What is Alport syndrome?]. PMID- 12557849 TI - Hospital index. PMID- 12557848 TI - [Physician's attitudes toward H. pylori infections]. PMID- 12557850 TI - Monitoring the occurrence of emerging forms of Creutzfeldt-Jakob disease in the United States. PMID- 12557852 TI - Effectiveness of risk communication. PMID- 12557851 TI - Neurologic manifestations of lightning strikes. PMID- 12557853 TI - Abstracts of the 10th International Congress of Medical Acupuncture and Related Techniques (ICMART). Edinburgh, United Kingdom, 4-6 May 2002. PMID- 12557854 TI - [Proximal femur fractures (Part 2)]. PMID- 12557856 TI - Special issue in honour of Peter W. Hawkes on the occasion of his 65th birthday in recognition of his contributions to electron optics and electron microscopy. PMID- 12557855 TI - Symposium on radionuclides in paediatric nephro-urology. PMID- 12557857 TI - Current awareness on yeast. PMID- 12557859 TI - Acute epiglottis in adults. AB - Acute epiglottitis can be a serious life-threatening disease because of its potential for sudden upper airway obstruction. It is a well-recognised entity in children but it is uncommon in adults and therefore is often misdiagnosed. In this retrospective study we present twelve cases of acute epiglottitis in adults. The diagnosis was made by visualisation of the epiglottis using fibreoptic laryngoscopy. The illness was managed using a standardised management protocol (see Appendix). The most frequent symptoms were odynophagia (100%), inability to swallow secretions (83%), sore throat (67%), dyspnoea (58%) and hoarseness (50%). Body temperature was elevated (>37.2 degrees C) in 75% and 50% of the patients had tachycardia (>100 bpm). The supposedly typical sign of stridor was found in only 42% of the cases. A routine oropharyngeal examination does not exclude epiglottitis, 44% of our patients had a normal oropharynx and the diagnosis could only be made following fibreoptic laryngoscopy. Nasotracheal intubation was necessary in four patients. A 40-year-old man with sore throat, hoarseness, cough and odynophagia was initially seen by a physician. With the suspected diagnosis of an infection - induced exacerbation of bronchial asthma, he was treated with antibiotics, paracetamol und corticosteroids. On admission six hours later the patient was in coma. The diagnosis was not made until conventional oral endotracheal intubation (without a tracheotomy set placed at the bedside) was attempted. Unfortunately the intubation failed and the patient died. Medical management of epiglottitis in adults includes an antibiotics, NSAIDs and possibly inhalation with adrenaline. The maintenance of an adequate open airway is the main concern in adults as well as in children. Although most adults have no signs of airway obstruction, the clinical threshold for insertion of an airway should remain low, as it is the only way of preventing death. A high index of suspicion is needed to recognise this rare disease correctly and patients must be admitted to a hospital with intensive care facilities, where the diagnosis can be confirmed and intubation performed if necessary and thus reduce the mortality rate. PMID- 12557858 TI - [Proceedings of the XXXIV Congress of the Polish Society of Internal Medicine. Poznan, 19-22 September 2001]. PMID- 12557861 TI - Proceedings of the Cell and Molecular Biology of Membrane Transport Systems and Disorders Conference. September 2-5, 2001. Coolangatta, Australia. PMID- 12557860 TI - Urban and rural origins of Otago medical students. AB - AIM: To investigate the home town population characteristics of medical students admitted to the Otago Medical School (OMS) in the context of recruitment for rural practice. METHODS: Data on home town population characteristics were collected from successful New Zealand applicants to the OMS between 1987 and 2000. Trends in the data across these years were examined, and the data compared to the wider New Zealand population. RESULTS: Of the students surveyed, 2353 responded to the questionnaire (98.4%). Of these, 2257 (94.4%) had identifiable home towns in New Zealand at the year before entry into the OMS. Of those with identifiable New Zealand home towns, 84.5% came from main urban areas, while only 2.9% came from rural areas, compared with 11.9% of the wider New Zealand population aged 17-27 years. The proportion of students in each of the urban/rural groups indicated only minor trends across the years surveyed, with the main urban group increasing and the rural and secondary urban groups decreasing. CONCLUSIONS: Between 1987 and 2000, in comparison with the wider New Zealand population, the pool of Otago medical students was disproportionately weighted towards highly populated, main urban areas and away from rural, low populated areas. The significance of the under-representation of rural students is discussed in relation to the recruitment and retention of rural doctors. PMID- 12557862 TI - The Alliance for Academic Internal Medicine: influence through size, diversity, and accountability. PMID- 12557863 TI - Screening for CADASIL mutations. PMID- 12557865 TI - Polymorphisms of adrenergic receptors and the risk of heart failure. PMID- 12557864 TI - Balloon angioplasty or medical therapy for hypertensive patients with atherosclerotic renal artery stenosis? A meta-analysis of randomized controlled trials. AB - PURPOSE: The optimal treatment for hypertensive patients with atherosclerotic renal artery stenosis is controversial. We performed a meta-analysis comparing the effects of balloon angioplasty and medical therapy in these patients. METHODS: We searched MEDLINE, EMBASE, the Science Citation Index, the Cochrane Controlled Trials Registry, and reference lists. Authors of published trials were contacted. RESULTS: We identified three trials involving a total of 210 patients with moderate-to-severe (> or = 50%) unilateral or bilateral atherosclerotic renal artery stenosis and poorly controlled hypertension who were followed for at least 3 months after intervention. Balloon angioplasty was significantly more effective in reducing blood pressure than was medical therapy; the weighted mean difference between the two treatments was -7 mm Hg (95% confidence interval [CI]: -12 to -1 mm Hg) for systolic blood pressure and -3 mm Hg (95% CI: -6 to -1 mm Hg) for diastolic blood pressure. There was no consistent difference in changes in renal function. Patients treated with balloon angioplasty were more likely to have patent renal arteries after 12 months (52% vs. 19%; odds ratio [OR] = 4.2; 95% CI: 1.8 to 9.8), used fewer antihypertensive medications, and appeared to have fewer major cardiovascular and renovascular complications (OR = 0.27; 95% CI: 0.06 to 1.23; P = 0.09). CONCLUSION: Balloon angioplasty has a modest but significant effect on blood pressure and should be considered for patients with atherosclerotic renal artery stenosis and poorly controlled hypertension. There is no evidence supporting its use in improving or preserving renal function, although none of the trials were designed to address this issue. PMID- 12557866 TI - Polymorphisms of adrenergic receptors and the risk of heart failure. PMID- 12557867 TI - Adolescent depression. PMID- 12557868 TI - Case 33-2002: a 28-year-old woman with ocular inflammation, fever, and headache. PMID- 12557869 TI - Case 33-2002: a 28-year-old woman with ocular inflammation, fever, and headache. PMID- 12557870 TI - Case 33-2002: a 28-year-old woman with ocular inflammation, fever, and headache. PMID- 12557871 TI - Clinical-trial agreements between medical schools and industry. PMID- 12557872 TI - Clinical-trial agreements between medical schools and industry. PMID- 12557873 TI - Perindopril effective in Africa-American hypertensives. PMID- 12557874 TI - One great leap for lipid lowering drugs. PMID- 12557875 TI - Progress and stroke--it's time to translate evidence into action. PMID- 12557876 TI - ASCOT--trial hypertensives with normal to elevated cholesterol levels. PMID- 12557877 TI - New B-type natriuretic peptide test (pro-BNP) for heart failure. PMID- 12557878 TI - Selective breeding program with rats: introduction. PMID- 12557879 TI - Bibliography. Current world literature. PMID- 12557880 TI - To be or not to be: the effects of aging stereotypes on the will to live. AB - This study examined whether stereotypes of aging might contribute to decisions the elderly make about when to die. Old and young participants (N=64) were subliminally primed with either negative or positive stereotypes of old age and then responded to hypothetical medical situations involving potentially fatal illnesses. Consistent with our prediction, the aged participants primed with negative stereotypes tended to refuse life-prolonging interventions, whereas the old participants primed with positive age stereotypes tended to accept the interventions. This priming effect did not emerge among the young participants for whom the stereotypes were less relevant. The results suggest that societally transmitted negative stereotypes of aging can weaken elderly people's will to live. PMID- 12557881 TI - The hot and the classic. PMID- 12557882 TI - How do the mental health issues differ in the withholding/withdrawing of treatment versus assisted death? AB - This article reviews ten of the most common mental health-related arguments against assisted death and applies them to the withholding/withdrawal of treatment to determine if the concerns expressed are truly unique to assisted death. After this analysis it is suggested that the mandatory involvement of a mental health professional can alleviate many of the concerns specifically associated with assisted death and that a mental health professional may also be helpful in situations involving the withholding/withdrawal of treatment. PMID- 12557883 TI - False expectations? Expectations vs. probabilities for dying. AB - It is widely recognized that the code of the physician has undergone dramatic changes in the last century--changes which have serious implications for the patient-physician relationship. This is an ethnographic study examining how certain changes in the role and abilities of biomedical physicians have affected patient attitudes and expectations about end-of-life care. In-home interviews were conducted with eighteen persons age fifty-five and older, including a sample of Hemlock Society members. Results indicate a broad spectrum of end-of-life concerns including capacity, autonomy, pain, and burden to loved ones. Most participants reported a reluctance to begin a discussion of death or future deteriorating capacity with their physicians. Instead, when conversations about death were reported, they had been largely limited to the scenarios of catastrophic illness (e.g., hospitalization, ventilator, etc.) and the Living Will. While this discussion does not overlook the utility of the Living Will, it proposes the reliance on this document for preparing patients for end-of-life care is inadequate. PMID- 12557884 TI - The good death: reaching an inductive understanding. AB - The study examines descriptions by twenty-six elderly Israelis of a good death. The transcripts of personal interviews were analyzed using phenomenological methods to determine general and essential essences of the good death phenomenon. The good death was perceived as a multidimensional phenomenon based on eighteen general essences that were condensed into five essential essences that included the physiological, personal, interpersonal, social and cultural domains of life. The good death description further involved a critical component toward the ways in which death and dying are currently occurring in Israel. The research results call for Israeli policy-makers to more forcefully acknowledge and accommodate the different secular perspectives of the good death into law and to allow individuals more freedom and control over the dying processes and rituals following one's death. PMID- 12557886 TI - Assisted suicide: factors affecting public attitudes. AB - Public support for assisted suicide has been growing despite the ethical questions raised by members of the medical profession. Previous research suggests that age, gender, experience, and religiosity are factors affecting individuals' attitudes. This study examines the effect of demographic and ideological factors, as well as individuals' caregiving experiences, on attitudes toward assisted suicide. Random-digit-dialing procedures produced a sample of 156 residents of Denton, Texas, in March 1998. T-tests were conducted to measure significance, while gamma values were used to measure level of association and percent reduction in error. The data indicate that age, gender, and caregiving experience were not significant predictors of attitudes. Situational factors, including whether a physician or friend/family member should assist and whether a child or a terminally ill patient experiencing no pain should receive assistance, all were highly significant and positively associated with attitudes toward assisted suicide. Respondents were most likely to support physician-assisted suicide for individuals experiencing no pain. The data also indicated that the depth of commitment to the beliefs that suffering has meaning, that life belongs to God, and that physician-assisted suicide is murder, were highly significant and negatively associated with attitudes toward assisted suicide. PMID- 12557885 TI - Older women and mercy killing. AB - Mercy killing is usually defined as intentional killing, often by family members or friends, with the stated intent to end perceived suffering. International evidence suggests that mercy killing typically involves an older man killing his ailing wife. In this study, we examined U.S. cases of mercy killing recorded by The Hemlock Society for the period 1960-1993. We found that the typical case involved an older woman being killed by a man, often her husband, with her poor health as the justification for the killing. A firearm was often used in these incidents. These patterns of mercy killing are consistent with patterns of homicide-suicide among older adults. Future research should seek to understand why women are typically the targets, and men the agents of mercy killing. PMID- 12557887 TI - On file. PMID- 12557888 TI - The conjoined twins Mary and Jodie: ethical analysis of their case. PMID- 12557889 TI - The Human Genome Diversity Project as a complement to human population genetics. PMID- 12557890 TI - The HGDP can be and should be pursued in harmony with its subjects: an account of the Israeli experience. PMID- 12557892 TI - It's more than the Human Genome Diversity Project. PMID- 12557891 TI - Intraventricular melanin-concentrating hormone stimulates water intake independent of food intake. AB - The lateral hypothalamus (LH) has a critical role in the control of feeding and drinking. Melanin-concentrating hormone (MCH) is an orexigenic peptidergic neurotransmitter produced primarily in the LH, and agouti-related protein (AgRP) is an orexigenic peptidergic neurotransmitter produced exclusively in the arcuate (ARC), an area that innervates the LH. We assessed drinking and eating after third ventricular (i3vt) administration of MCH and AgRP. MCH (2.5, 5, and 10 micro g i3vt) significantly increased food as well as water intake over 4 h when administered during either the light or the dark portion of the day-night cycle. When MCH (5 micro g) was administered to rats with access to water but no food, they drank significantly more water than when given the vehicle. AgRP (7 micro g i3vt), on the other hand, increased water intake but only in proportion to food intake during the dark and the light, and water intake was not increased after i3vt AgRP in the absence of food. Hence, in contrast to AgRP, MCH elicits increased water intake independent of food intake. These results are consistent with historical data linking activity of the LH with water as well as food intake. PMID- 12557893 TI - The overlooked ethics of the Human Genome Diversity Project. PMID- 12557894 TI - Is there a need for the HGDP, now that the HGDP is interested in human variation? PMID- 12557895 TI - The devil, the details, and the HGDP. PMID- 12557896 TI - Human population genetics versus the HGDP. PMID- 12557897 TI - The HGDP in the best possible world. PMID- 12557898 TI - Ethics and the Human Genome Diversity Project: an African perspective. PMID- 12557899 TI - Time changes all things, yet there is nothing new under the sun: the transience of ethical norms. PMID- 12557901 TI - Bibliography. Current world literature. PMID- 12557900 TI - Retraction: Universal Crossover from Band to Hopping Conduction in Molecular Organic Semiconductors [Phys. Rev. Lett. 86, 3843 (2001)]. PMID- 12557902 TI - [Anatomia practica: features from the history of early patho-anatomy]. AB - Since the anatomy school of Alexandria during the fourth og third century before Christ dissection of the human body seems not to have been practiced until late Medieval or early Renaissance period, undoubtedly due to ethical and religious aversions. The teaching of anatomy was based on Galen using animal dissection. In the fourteenth and fifteenth centuries, however, anatomical examinations of the human body slowly began, seemingly for the purpose of describing both the normal structure and the abnormal structure caused by diseases, maldevelopment or trauma. This latter branch of anatomy was called practical, medical or correlative anatomy and corresponds to what we today name as patho-anatomy. Antonio Benivieni of Florence (1442-1502) is the first one to collect (and publish) a series of clinical observations some of which could be correlated to post mortem findings. It is unknown, however, whether the autopsies were performed by himself; and there is no mentioning of technique or circumstances for sectioning. Studies of the dead body by incision for the purpose of displaying diseased organs (autopsy) seem to have been an accepted practice for which relatives consented in those days. Other medical doctors in the years to follow, as for instance Fernel (1485-1558) in Paris, Eustachius (1524-1574) in Rome, Felix Plater (1536-1614) in Basle and Th. Bartholin (1616-1680) in Copenhagen have used the anatomical method for the study of diseases. Further, Schenck (1530-1598) in Freiburg and Bonet (1620-1689) in Geneva collected and published large series of clinical symptoms which had been related to post mortem findings dating back to ancient observers. This is the scientific background for anatomists as Morgagni, Lieutaud, Baillie, Bichat and others who founded the morbid anatomy on which the study of disease flourished in the classical patho anatomical era of the nineteenth century with names as Rokitanski and Virchow. PMID- 12557903 TI - Gene targeting in immunology. AB - Transgenics and gene targeting by homologous recombination provide an ideal opportunity to delineate immune functions of specific genes. These gene knockout mice are powerful tools to understand complex mechanism of immune system and molecular processes underlying autoimmune diseases and immunodeficiencies. Availability of an increased pool of genetically manipulated mice will provide a basic foundation for developing of novel strategies to treat immunological diseases. PMID- 12557904 TI - Poly (DL-lactide-co-glycolide) based delivery systems for vaccines and drugs. AB - Current vaccination and drug delivery strategies emphasize on the development of controlled release techniques for persistent and sustained effects. In the recent years, polymer based systems for the delivery of bioactive agents have gained considerable attention due to their marked adjuvanticity, established biodegradability and biocompatibility, excellent mechanical strength and controlled release profiles. This review deals with the potential applications of synthetic polymers mainly PLG polymers in delivery of vaccines and drugs. PMID- 12557905 TI - Signal transduction by xenobiotics in fish. AB - Signal transduction by xenobiotics in fish has recently gained much attention. The better known transduction mechanisms are those elicited by organochlorines, organophosphates, carbamates and heavy metals. Organochlorines specifically bind to the membrane bound ouabain sensitive Na+-K+-ATPase affecting neural transmission while the organophosphates and carbamates bind specifically to the membrane bound enzyme acetylcholinesterase again affecting neural transmission. Since the nervous system is one of the important integrative and interactive physiological systems in animals, hypofunction of the nervous system leads to secondary effects in the endocrine system including thyroidal, gonadal, interrenal, pituitary and hypothalamic functions. Even low levels of xenobiotics are efficient enough to bring about remarkable changes in the functional physiology of the non target animals. Heavy metals such as cadmium or mercury belonging to the same group II B in the periodic table probably have a similar mechanism of action. Avidity of these metals to SH-radicals allow them to bind indiscriminately to SH groups in proteins. One pathway of interaction by inorganic mercury with the membrane bound ouabain sensitive Na+-K+-ATPase has been clearly established in fish liver and ovary. Binding of inorganic mercury to the membrane bound enzyme is through sulfhydryl group which inactivates the sodium pump leading to accumulation of the cation in the cytosol. The inorganic mercury is next conjugated by the cytosolar nucleophile, glutathione, and is transported to the nucleus where dissociation occurs and the free metal binds to the metal regulatory element to initiate gene expression. The inducible proteins are 3beta-hydroxysteroid dehydrogenase in the oocyte and metallothionein and C reactive protein in the liver. The present review deals with the role of xenobiotic as a stress factor. PMID- 12557907 TI - On speed and aerodynamic forces of mosquito. AB - In the present investigation, speed of mosquitoes A. aegyptii (Linne) and Culex quinquefasciatus (Say) is measured by designing and constructing a low speed wind tunnel in the laboratory. The velocity of mosquitoes is less than the other myogenic and neurogenic insects. Lift, one of the important aerodynamic forces that a flier has to develop for its efficient flight, is studied in mosquitoes by developing a simple technique using digital single pan balance. Lift, drag and their coefficients of hovering mosquito are calculated from the knowledge of body parameters by considering the wings of mosquito as harmonic oscillator. The calculated value of lift is verified with the experimental. The study throws light on morphophysiological adaptation of mosquitoes for the generation of aerodynamic forces in hovering, tethered and forward flights. PMID- 12557906 TI - Intestinal cholesterol synthesis and mobilisation to blood in sucrose-feeding and alcohol-intake. AB - Small intestines synthesize cholesterol to a greater extent than liver. Between starch-fed and sucrose-fed rats, using (14C) glucose, it was found that the synthesis of cholesterol by the jejunum of small intestines was greater in the sucrose-fed group than starch-fed group. By a novel experimental technique and using (14C) glucose, it was found that the contribution towards buffer representing lymph was greater in the sucrose-fed group (13.3%) than the controls (11%). Hypercholesterolemia on sucrose feeding may be at least partly due to contribution by the small intestines. Regarding alcohol, using (14C) glucose it was found that total synthesis of cholesterol by the small intestines was decreased in alcohol-fed rats. There was no difference in the cholesterol retained by the intestinal tissue between the controls and alcohol-fed animals while, the secretion towards buffer (lymph) was 9% as against 11. This indicates that there is contribution of cholesterol to blood from small intestines in alcohol-intake also but due to overall decrease in the intestinal synthesis of cholesterol, contribution of intestines to hypercholesterolemia may not be substantial as in the case of sucrose feeding. This is because in sucrose-feeding there is increased cholesterol synthesis. PMID- 12557908 TI - Electrophysiological studies with repeated episodes of ischaemia on isolated rat heart. AB - In order to know the beneficial effect of preconditioning electrocardiography recording were used as tool to assess myocardial malfunction and for this perfusion apparatus was setup. Electrophysiological changes for each heart were recorded during perfusion at 1, 2, 3, 5, 10, 20, 30 and 60 min of global ischaemia and also during the equal period of reperfusion. Recordings dembnstrate that the normal rate was about 240 beats/min with an "R" amplitude of 4mV. During the first ischaemic episode of 1min the rate was 180 +/- 15 beats/min (counted as per 'R' wave deflection), at 2 mins it was 60 +/- 6 beats/min, at 3 min the rate was 40 +/- 2 beats/min, at 5 mins of ischaemia it was 90 +/- 6 beats/min, at 10 min 20 +/- 2 beats/min, at 20 min the rate was 60 +/- 4 beats/min, and at 30 mins there were nil beats/min. The recovery during all the periods of reperfusion was restored to between 120 and 180 beats/min in all episodes. Further after a 60 min of ischaemia the heart stopped to elicit any mechanical response. It is concluded that short term ischaemia can induce a resilient effect on the beating of the heart after a few episodes as seen subsequent to 1 and 2 min of ischaemia. Further, preconditioning was beneficial up to 30 min, beyond which the heart showed signs of fatigue and irreversible injury. PMID- 12557909 TI - Effect of excessive intake of thermally oxidized sesame oil on lipids, lipid peroxidation and antioxidants' status in rats. AB - Fresh and thermally oxidized sesame, groundnut and coconut oils were fed to different groups of rats, as high fat diet (20%). Feeding fresh and thermally oxidized oils increased the levels of total cholesterol, low density lipoprotein cholesterol (LDL-C), and phospholipids but high density lipoprotein cholesterol (HDL-C) decreased in all the experimental animals. The levels of very low density lipoproteincholesterol (VLDL-C) and triacylglycerol increased only in groundnut and coconut oils-fed groups and decreased in sesame oil-fed group when compared with the control. When fresh and the corresponding thermally oxidized oils-fed groups were compared with the control, total cholesterol and LDL-cholesterol alone increased while triacylglycerol, VLDL-cholesterol, HDL-cholesterol, HDL/LDL ratio and phospholipids decreased. Thiobarbituric acid reacting substances increased in all the experimental animals and more so in corresponding thermally oxidized oils. It was less pronounced in sesame oil-fed groups when compared with the corresponding other oils-fed groups. Feeding of thermally oxidized oils decreased the levels of vitamin E, vitamin C and reduced glutathione when compared with fresh oils. Among the three thermally oxidized edible oils, sesame oil exhibited lesser risk for hyperlipidemic disorders. PMID- 12557910 TI - Hypoglycemic effect of aqueous extract of Enicostemma littorale Blume (chhota chirayata) on alloxan induced diabetes mellitus in rats. AB - The whole plant aqueous extract of E. littorale was tested for its hypoglycemic activity on normoglycemic, hyperglycemic and alloxan induced diabetic rats. Blood sugar lowering activity was not observed in normoglycemic and glucose loaded hyperglycemic rats in the short time experiment. But in case of diabetic rats, the fall of blood sugar after 30 days treatment with the aqueous extract was found to be significant (P < 0.001). The decrease in the plasma glucose level was accompanied with decrease in the level of glycosylated haemoglobin and glucose-6 phosphatase activity in liver. The potent anti-diabetic properties of E. littorale has been reported for the first time. PMID- 12557911 TI - Comparative effectiveness of Tiron (4,5-dihydroxy benzene 1,3-disulphonic acid disodium salt) and CaNa2EDTA with time after beryllium poisoning. AB - The efficacy of two chelating agents (Tiron and calcium disodium EDTA) in the treatment of beryllium induced blood biochemistry and hepatic histopathological alteration was investigated at different duration in female albino rats. Single administration of beryllium nitrate at a dose of 50 mg/kg (im) showed significant decrease in haemoglobin percentage, blood sugar level, protein contents and activity of alkaline phosphatase. On the contrary significant elevation was found in the activity of transaminases (AST and ALT). Tiron was found to be more effective than CaNa2EDTA in reducing the beryllium induced haematological alterations and histopathological lesions in liver. These findings were further confirmed by AAS thus, in which reduced beryllium body burden was seen in liver and blood with Tiron. PMID- 12557912 TI - Seroreactivity of purified Brugia malayi microfilarial soluble and excretory secretory antigens in different clinical presentations of bancroftian filariasis. AB - Brugia malayi microfilarial excretory-secretory (mf ES) and phosphate buffer saline soluble (mf S) antigens were fractionated by fast protein liquid chromatography (FPLC) on superdex 200 HR 10/30 gel filtration column. The active antigen fractions were identified and explored in comparison with whole mf ES and mf S antigens to detect filarial IgG antibodies in different groups viz microfilaraemics, acute, chronic and occult filarial cases of Wuchereria bancrofti infection and endemic and non-endemic normals. One of the fractions of mf ES antigen (ESF-6) and two fractions of mf S antigen (SF-2 & 3) were identified to be useful to detect filarial antibodies. A pooled preparation of these antigen fractions gave a sensitivity of 86.6% (for microfilaraemic cases) and a specificity of 95% to detect filarial IgG antibodies by indirect ELISA. The pooled FPLC purified mf antigens also showed 55-88% of cases of different grades of clinical filariasis and 65% of tropical pulmonary eosinophilia cases as positive for filarial antibodies. The pooled FPLC purified B. malayi mf antigens with higher specificity are preferable to whole mf ES and mf S antigens to detect active filarial infection in microfilaraemia and as well in different clinical entities of bancroftian filariasis. PMID- 12557913 TI - Variability of morphological traits in Drosophila bipectinata complex. AB - Phenotypic or morphological differences among different populations and sexual dimorphism in certain metric traits were analysed in D. bipectinata complex. It was noticed that different populations of D. bipectinata species group harbour large amount of variation for these characters. In all the populations, morphometric characters such as lengths of femur, tibia and wing length, wing width, number of sternopleural bristles and bristles on epandrium varied significantly among populations. The study indicates that the morphological variations are due to the interplay of genetic and environmental endowments. Further, females had significantly larger values, for lengths of femur, tibia and wing length, wing width and sternopleural bristles. PMID- 12557914 TI - Blue light-sensitive plasma membrane bound exogenous NADH oxidase in Cuscuta reflexa. AB - Protoplasts isolated from Cuscuta reflexa exhibited a higher rate of exogenous NADH oxidation as compared to NADPH in the dark. NAD(P)H oxidation was monitored by measuring the rate of oxygen consumption and this oxidase system was sensitive to blue light. Both NADH oxidase and its blue light sensitivity were inhibited by -SH group reacting agents. The corresponding changes occurring in H+-extrusion activity and intracellular ATP levels were also monitored. Stimulation of NADH oxidation under blue light corresponded to increased rate of H+-extrusion and intracellular ATP level, the converse was also true under NADH oxidase inhibitory conditions. These observations suggested a close functional association between blue light-sensitive plasma membrane bound redox activity and H+-ATPase in this tissue. Further, concanavalin A binding of protoplasts resulted in a loss in NADH oxidase activity and its blue light sensitivity suggesting apoplastic location and glycoprotein nature of the blue light sensitive NADH oxidase system in Cuscuta. PMID- 12557915 TI - Accumulation of lead and cadmium in upper parts of mustard (Brassica juncea) seedlings in response to putrescine. AB - Lead and cadmium accumulation examined in shoot and leaf tissues of seedlings of mustard (Brassica juncea cv RH-30), at 7th day, treated with either putrescine (1 mM), or ammonium nitrate (10 mM) or IAA (10 microM). These were included in the nutrient medium, containing Pb or Cd (0.1 mM and 2 mM). Metal accumulation was more in shoot than in leaf tissues, which was increased manifold under saline conditions. However, Cd accumulation in tissues was higher than Pb. Chemical (putrescine, ammonium nitrate or IAA) treatment of the seedlings, decreased metal accumulation in leaf (10-20%) and in shoot (40 to 60%) tissues, depending upon external metal levels. Putrescine significantly decreased the metal accumulation and translocation under saline conditions. PMID- 12557916 TI - Influence of cadmium on growth and development of Vicia faba Linn. AB - Influence of cadmium (Cd) on growth and development of broad bean (V. faba) was assessed in pot cultures with cadmium iodide (CdI2) in different concentrations ranging from 15 to 500 mg per kg of soil. There was a decline in plant height and total dry weight. Root size decreased most significantly with a corresponding reduction in the frequency of root nodules. Total soluble protein in leaf, stem and root suffered a pronounced loss with increasing concentration of cadmium. Chlorophyll a was the most sensitive pigment followed by chlorophyll b and carotenoids. Nitrate reductase activity too was adversely affected. Cadmium contamination induced abnormalities in stomata and trichomes. PMID- 12557917 TI - Suitability of limited sampling strategy to cyclosporine A oral formulation. AB - Therapeutic drug monitoring of cyclosporine A in renal transplant patients is normally performed by measuring pre-dose trough levels or doing area under the curve profiles from all possible sampling points. On the basis of good correlation between predicted area under the curve from limited sampling points and actual area under the curve (literature survey), a limited sampling strategy at 1, 3 and 5 hr was used and extended this strategy as a criteria for pilot evaluation of modified cyclosporine A formulations. Hemodialysis patients served as subjects for the study. Panimun Bioral was used as a test formulation while Sandimmun Neoral as standard reference. The study revealed that both formulations had similar restricted AUC0-5h profiles. PMID- 12557918 TI - Effect of abana an ayurvedic formulation, on lipid peroxidation in experimental myocardial infarction in rats. AB - The present study was conducted to elucidate the antioxidant role of an ayurvedic formulation Abana in isoproterenol induced myocardial infarction in rats. In myocardial necrosis induced by isoproterenol, a significant increase in serum iron content with a significant decrease in plasma iron binding capacity, ceruloplasmin activity and glutathione level were observed. There was also a significant increase in lipid peroxides levels on isoproterenol administration. Activities of antioxidant enzymes like superoxide dismutase, catalase, glutathione peroxidase, glutathione-s-transferase, glutathione reductase were decreased significantly in heart with isoproterenol-induced myocardial necrosis. Abana, produced a marked reversal of these metabolic changes related to myocardial infarction induced by isoproterenol. In conclusion ayurvedic formulation Abana exerts its effect by modulating lipid peroxidation and enhancing antioxidant and detoxifying enzyme systems. PMID- 12557919 TI - Effect of cold pressor test on visual reaction time and auditory reaction time. AB - Influence of stress (cold) on visual reaction time (VRT) for red, green and yellow colours and auditory reaction time (ART) for high and low pitched sounds was studied in 60 healthy subjects (17-19 years) by digital display response time apparatus. Pain threshold and pain tolerance time were 23.02 +/- 8.11 and 249.35 +/- 64.88 sec. respectively. During the standard cold pressor test (CPT), there was significant increase (P < 0.01) in VRT for all the three colours, the increase being the least for the red colour. There was significant increase (P < 0.01) in ART for both high and low pitched sounds. There was increase in heart rate and respiratory rate as compared to controls. This increase in VRT and ART with CPT was probably due to decreased conduction velocity of nerves, which could be due to vasoconstriction attributed to increased sympathetic activity caused by cold and pain. PMID- 12557920 TI - Effect of tonic pain on schedule specific feeding behaviour. AB - Food deprivation produces analgesia. This response is reversed i.e. pain sensitivity is lowered, when the food deprived rats are fed. In the present study the effect of chronic pain on the motivation to get food, in food deprived rats, was observed. In ten rats the effect of formalin and morphine plus formalin on the motivation to get food was studied. Injection of formalin significantly (P < 0.01) decreased the number of lever presses from 450 +/- 30 to 225 +/- 25. However, after injecting morphine the effect was reversed. The present study shows reduced internal drive to procure food by the food deprived animals, when they were under chronic pain. The effect was blocked by morphine, suggesting the role of opioids in modulating the motivation for getting food. PMID- 12557921 TI - Influence of ethanolic extract of Tephrosia purpurea Linn. on mast cells and erythrocytes membrane integrity. AB - The ethanolic extract of T. purpurea Linn. was studied for its in vitro effect on rat mast cell degranulation and erythrocyte membrane integrity in vitro. The extract in concentration of 25-200 microg/ml showed a dose-dependant inhibition of rat mast cell degranulation induded by compound 48/80 and egg albumin. T. purpurea extract was found to inhibit haemolysis of erythrocytes induced by hypotonic solution but accelerated haemolysis induced by heat at a concentration of 100 microg/ml. The studies reveal that the ethanolic extract of T. purpurea may inhibit degranulation of mast cells by a mechanism other than membrane stabilization. PMID- 12557922 TI - Photosynthesis and kinetic characteristics of rubisco in Hibiscus cannabinus L. AB - Photosynthetic characteristics in kenaf (Hibiscus cannabinus L.), a C3 plant, were compared with Abelmoschus esculentus (L.) Moench, another member of Malvaceae. Kenaf leaves exhibited significantly higher rate of photosynthesis (40 mg CO2 dm(-2) hr(-1)) which was 24.6 mg dm(-2) hr(-1) in A. esculentus. Rate of photo and dark respiration was similar in both the species. Kenaf leaf photosynthesis had a higher optimum temperature (32 degrees C) than that of A. esculentus (26 degrees C). Photosynthesis in kenaf leaves required higher saturation irradiance (1,600 micromole m(-2) sec(-1)). There was a significant correlation between photosynthetic rate and biomass yield in these species. The primary product of photosynthesis after 5 seconds of 14C-assimilation was 3-PGA in both the species. The kinetic properties of RuBP carboxylase/oxygenase were determined in the leaf extracts. Higher carboxylase activities were recorded with kenaf leaf extracts (245 pmole mg chl(-1) hr(-1)). Km (CO2) for kenaf leaf carboxylase was significantly lower (7.8 microM) than A. esculentus (13.5 microM) and corresponding difference in Vmax values of carboxylase was recorded between the two species. The kinetic characteristics of oxygenase were similar in both the extracts. These results indicated the variation in carboxylase activity and its kinetic characteristics reflected a significant difference in CO2 assimilation in C3 plants. PMID- 12557923 TI - Biotransformation of banana waste into protein by pleurotus sajor-caju. AB - Pseudostems and leaves from banana waste were used for biotransformation into protein by using P. sajor-caju, an oyster mushroom. Treatment of formalin (500 ppm) + carbendazim (12.5 ppm) of these substrates was found to favour relatively high percentage biological efficiency (BE) of P. sajor-caju.. Steam sterilization also exhibited comparable yield performance by P. sajor-caju. Fruiting bodies harvested from all the treatments had relatively higher protein contents. The spent substrate (steam sterilized) was found to be suitable as an ideal animal feed because of its rich nutritive composition. PMID- 12557924 TI - Fine structure of the synganglion of unfed female Rhipicephalus (Rhipicephalus) sanguineus (Latreille) (Ixodoidea: Ixodidae). AB - The synganglion of unfed female Rhipicephalus (Rhipicephalus) sanguineus (Latreille) is enclosed within a well defined neural lamella underlined by the perineurium. No organelles were observed in the neural lamella, whereas axons containing neurosecretory vesicles, tracheal elements and glycogen deposits are visible in the perineurium. Below the perineurium, a cortical layer of nervous and non-nervous elements surrounds an inner neuropile, the latter is enclosed by a thin subperineurial layer. The neuropile is formed of nerve fibers containing neurotubules. PMID- 12557925 TI - Morphometric and histochemical study of the adrenal cortex in experimental schistosomiasis mansoni. AB - In this study, 45 mice were used, 30 mice of them were injected subcutaneously with 80-100 schistosoma mansoni cercariae and 15 mice remained uninfected and served as a control group. Infected mice were killed at 8 and 12 weeks post infection while control mice were killed at one time. Paraffin sections were made from adrenal cortex and liver. The thickness and cellularity of zonae glomerulosa and fasciculata as well as the serum cortisol level were determined. In the infected group of mice, the zona glomerulosa and fasciculata increased in the thickness and cellularity and the serum cortisol level increased with the progression of infection as compared with the control uninfected group. Bilharzial liver granulomas were profound in the infected group at 12 weeks post infection which coincided with the time of suprarenal hyperactivity. PMID- 12557926 TI - Laboratory evaluation of a Helicosporidium sp. (Protozoa: Helicosporida) as an agent for the microbial control of mosquitoes. AB - A Helicosporidium sp. isolated from diseased larvae of Culex pipiens L. in Egypt was assessed for its microbial control potential against mosquito larvae. Dose infectivity and dose mortality of the pathogen was determined for each larval instar of Cx. pipiens. Helicosporidium sp was infectious to Aedes caspius (Pallas), C. antennatus (Becker), C. perexiguus Theobald, and Culiseta longiareolata (Macquart). Using 24-hr exposures of first instar larvae to a range of concentrations between 50 and 5 x 10(3) spores/ml, Cx. antennatus and Cx. perexiguus were more susceptible than Cx. pipiens, Ae. caspius and Cs. longiareolata with LC50's of 5 x 10(2), 5 x 10(2), 1.6 x 10(3), 1.4 x 10(3), and 1.7 x 10(3), respectively. Current experiments shown that Helicosporidium spores were tolerant of freezing and storage at 5 degrees C, but not to desiccation. Production trails indicated that up to 6.2 x 10(8) spores could be routinely produced in individual cotton leaf worms, Spodoptera littolaris (Boisd) (Lepidoptera: Noctuidae). PMID- 12557928 TI - The prevalence of lice infesting students of primary, preparatory and secondary schools in Cairo, Egypt. AB - Human lice (head and body) are among the arthropod-ectoparasites of worldwide distribution. Examining students in primary, preparatory and secondary schools recorded prevalence rates of 21.86%, 30.38% and 12.94% respectively. The overall rate of the lice infestation in the three schools was 384 out of 1772 or 21.67%. The prevalence rate of lice infestation among males and females were 17.02% & 37.8% (primary school), 27.8% & 33.1% (preparatory school), and 12.0% & 13.9% (secondary school). These totaled 17.7% (males) and 30.26% (females). The overall ratio of head to body lice was 18.2:1. Consequently, lice mainly the head louse, are still a public health problem particularly among female students in the primary and preparatory schools. In the secondary school prevalence rate of the lice infestation was low. So human lice is still a community health problem. PMID- 12557927 TI - Evaluation of the role of some parasitic infections as a cause of acute and chronic hepatic diseases. AB - A total of 200 patients with acute and chronic hepatic diseases and 100 apparently normal cross-matched controls were examined for the parasitic cause. The IHAT showed 3% had hydatid cysts, 3% had fascioliasis and 53.5% had schistosomiasis. However, no eggs were detected in the fascioliasis patients and only 4 & 3 cases showed S. mansoni and S. haematobium eggs in stools & urines respectively. ELISA showed 10% with visceral larva migrans & 65.5% with toxoplasmosis (IgG, IgM or both). On the other hand, 27% of the controls showed antibodies against toxoplasmosis. PMID- 12557929 TI - Genital system structure of unfed female Rhipicephalus (Rhipicephalus) sanguineus (Latreille) (Ixodoidea: Ixodidae). AB - The reproductive system of female Rhipicephalus sanguineus consists of a single ovary, paired oviducts fusing into a single common oviduct, connecting tube, the vagina that in turn divides into cervical and vestibular regions, a wide receptaculum seminis and tubular accessory glands that open into the vagina between the cervical and vestibular regions. The histological structure of each part of the female reproductive system is described. The fine structure of the ovary, connecting tube and accessory glands is also provided in this study. PMID- 12557930 TI - Experimental studies on cyclosporiosis. AB - Swiss albino mice get Cyclospora infection after orally inoculated with sporulated oocysts. Two weeks post inoculation, most of them passed numerous acid fast immature oocysts in their stools. One week later, light microscopic examination of their intestinal H & E stained sections revealed parasitic stages in a supranuclear location within enterocytes. They were most prominent in the mucosal villi. Lamina propria was expanded by an inflammatory infiltrate. The combined parasitological and histopathological present studies helped in distinguishing Cyclospora which is often confused with Cryptosporidia in stool samples and with Isospora in intestinal sections. Electron microscopy demonstrated both sexual and asexual developmental stages of Cyclospora in ultrathin infected sections. Therefore, Cyclospora species require only a single host to complete its entire life cycle PMID- 12557931 TI - The effect of ivomec super on dogs experimentally infested with Sarcoptes scabiei isolated from a patient. AB - Scabies infestation is a community health problem worldwide. This is particularly true in overcrowded and unhygienic areas. Seven dogs were experimentally infested with Sarcoptes scabiei freshly recovered from a patient. The dogs after infestation were treated with a mixture of ivermectin and clorsulon (Ivomec Super). The dose given to a single dog was 1 ml/50 kg body weight. This is equivalent to 200 mcg ivermectin and 2 mg clorsulon per Kg body weight. Five of the dogs (71.43%) were completely cured with 1 ml/50 Kg body weight. The remaining two dogs needed each, another injection of the same dose two weeks later. So, ivomec super is effective against the human strain of Sarcoptes scabiei in experementally infested dogs. A general discussion was given. PMID- 12557932 TI - Studies on the megatrypanum trypanosomes of the Egyptian bat (Pipistrellum kuhli) from Sohag Governorate, Egypt. AB - Out of 150 Egyptian bats (Pipistrellus Kuhli) hunted in the premises of Sohag City, 60 (40%) were harbouring typanosomes of the subgenus Megatypanum (Hoare, 1964). They were T. M. heybergi (Rodhian, 1923), T. M. possoai (Dean and Sugary, 1963) and T. M. magadermae (Wenyon, 1990). PMID- 12557933 TI - Role of anti-oxidant preparation (oxi-guard) on Cryptosporidium parvum infection in albino mice. AB - Oxi-guard is a commercial anti-oxidant preparation in the form of capsules. One hundred and twenty albino mice were divided into three groups, first (50 mice) received oxi-guard for one week before infection with Cryptosporidium oocysts and continued for 50 days post-infection (P.I.), second (50 mice) did not receive oxi guard and infected with Cryptosporidium oocysts and third (20 mice) was neither infected, nor receiving oxi-guard, a control group. Histopathological examination of small intestines and counting of endogenous stages of Cryptosporidium in the epithelial surface were done. Malonyldialdehyde (MDA) levels in small intestines were measured in all mice. It had been observed that the administration of oxi guard led to significant increase in the numbers of endogenous stages of Cryptosporidium in the small intestines of group (1) in comparison with group (2). MDA levels showed a significant increase on the 15th, 20th and 25th days P.I. in group (2) and they did not show any significant changes in group (1) in comparison with the control group. As regards the activity of endogenous anti oxidant enzymes in Cryptosporidium parvum oocysts, there was a significant increase in superoxide dismutase in group (2) of mice compared with its level in group (1). Finally, it can be concluded that the administration of oxi-guard had led to eradication of free radicals which in turn prevented their lethal effects on the parasite leading to increased parasitic colonization in the small intestine of mice. PMID- 12557934 TI - Control of Musca domestica third instar larvae by the latex of Calotropis procera (Family: Asclepiadaceae). AB - Calotropis procera (Family: Asclepiadaceae) is known to contain alkaloids, steroids and resinous substance. Different concentrations of its latex were topically applied to the 3rd stage larvae of Musca domestica. A dose of 3 ul (5% of the latex) killed and partially digested the larvae in three hours. The effect of this latex is most probably enzymatic in nature. No doubt, the shift to plant extracts in controlling insect-vectors or pests in general, pave the way to a health environment. PMID- 12557935 TI - Evaluation of a possible role of tissue inhibitor of metalloproteinases 1 (TIMP 1) in the pathogenesis of hydatid disease. AB - Tissue inhibitor of metalloproteinases-1 (TIMP-1), in the serum levels of 27 patients was significantly higher and almost a double of that measured in healthy patients. Considerably higher TIMP-1 levels were recovered in patients with higher specific antibody titers and those with hepatic cysts in relation to low titer patients and pulmonary cysts respectively, reflecting an immunologic dose dependent and cell type-specific regulation of TIMP-1 production. Yet, these higher TIMP-1 levels stop short to be statistically significant in both groups. Patients with multi-sited hydatid cysts also show statistically insignificant higher TIMP-1 levels compared to single-sited cysts. PMID- 12557936 TI - Pattern and cost of medical care for workers with schistosomiasis. AB - This study describes the pattern of medical care provided to workers with schistosomiasis, estimate the total medical cost and to identify the proportional rates of sickness retirement attributed to schistosomiasis. The observational approach was adopted for this study 170 schistosomiasis workers and a similar number of controls were included in this study. An interviewing schedule and a special format were designed for collecting personal, medical and early retirement data. The results revealed that the mean total cost in the outpatient clinics was significantly higher for schistosomiasis workers than their controls (320.2 " 330.11 versus 210.8 " 260.01 L.E). The hospital cost was also higher for schistosomiasis workers compared with their controls (265.9 " 674.47 vs 195.8 " 629.72 L.E) but this differencewas not statistically significant. More than 80% of the total hospital cost was spent on bed cost. The average operative cost/worker was significantly higher among the schistosomiasis workers than the control workers (7.08 " 22.07 vs 2.35 " 5.2 L.E). The total medical cost (outpatient and hospital) was significantly higher for workers with schistosomiasis compared with their controls (586.02" 845.77 vs 406.57 " 694.34). The total number of workers who retired because of sickness disability other than schistosomiasis increased from 1994 to 1998 with a ratio of 2.54 while those who retired because of schistosomiasis and its complications increased with a ratio of 3.64. PMID- 12557937 TI - Study on the possible role of heat shock proteins in host resistance to Trichinella spiralis infection in experimental animals. AB - The expression of heat shock proteins (HSPs) and their immunogenic role in host parasite relationship has been studied in T. spiralis infection in mice. Two groups of infective stage larvae were incubated at 37 degrees C or at 43 degrees C for 3 hours in a nutrient medium. Crude somatic extracts and excretory/secretory (E/S) products were obtained. Quantitative estimation and in vitro translation of mRNA were carried out. Crude somatic extracts, E/S products and in vitro translated proteins were all subjected to SDS-PAGE and immunoelectrophoresis against a monoclonal antibody to HSP70. They were further used to immunize mice which were then infected with T. spiralis. The degree of immunization was assessed by counting adult worms, muscle larvae and estimating total gamma globulins of mice. SDS-PAGE revealed intense peaks of 71 KDa and 81 KDa in all proteins obtained from heat shocked larvae. These proteins have been proved to involve HSP70 as manifested by their strong reactivity with anti-HSP70. In vitro translated products of heat shocked larvae (at 43 degrees C) proved to be strongly immunogenic as evidenced by the lower adult worm, muscle larval counts and higher total gamma globulins in sera of mice in comparison with the control non immunized group as well as to other larval antigens incubated at 37 degrees C. So, exposure to heat, can induce the synthesis of heat shock proteins which can defend the organism against environmental stress. Meanwhile, these heat shock proteins can render the host more refractory to reinfection. PMID- 12557938 TI - The effect of ecological parameters on the distribution of snail vectors of schistosomiasis. AB - The infestation of the water courses showed 32.5% for Biomphalaria alexandrina and 8.75% for Bulinus truncatus. Ecological parameters, showed non significant variations in the water courses harbouring snail vectors and those free from snails except for conductivity in the habitats harbouring B. truncatus. This variation was more highly significant (p<0.001). Of the examined sites, 11.25% were harbouring B. alexandrina and Lymnaea natalenesis living together and 5% of the sites were harbouring B. truncatus and Physa acuta snails. Snail vectors were distributed with different degrees with aquatic plants reflecting the degree of species preference plants for snails' life. PMID- 12557939 TI - Impact of type and stage of schistosomiasis on quality of life and productivity of infected workers. AB - All workers with schistosomiasis (205 workers) during the field period (April 1998 to March 1999) were included. The study tools included an interviewing schedule, the WHO QOL-brief and a production data format specially designed to obtain data about the productivity of the workers. The data revealed that 78.8% of the schistosomiasis workers suffered from Schistosoma mansoni while only 7.6% suffered from S. haematobium and 13.5% had a mixed infection. Workers infected with S. mansoni tended to have lower mean percent of all the QOL domains and productivity indicators compared with workers infected with S. haematobium but these differences were not statistically significant. Workers with complications of schistosomiasis had significantly lower values for the QOL physical, social relationship and environmental domains (55.27 +/- 15.59, 74.94 +/- 5.34 and 56.39 +/- 15.09 versus 61.38 +/- 5.74, 79.36 +/- 11.96 and 61.64 +/- 7.74 respectively). Moreover, the production score was significantly lower in the workers with complication compared with workers free from schistosomiasis complications. The mean percent of QOL domains decreased as the stage of schistosomiasis increased from stage 1 to stage 4, this was statistically significant for the physical and independence domain. The mean additional hours and production scores decreased successively from stage 1 to stage 4 of schistosomiasis. Thus, special attention should be paid to schistosomiasis workers for early detection and treatment of complications to improve the QOL and increase the productivity of those workers. The impact of the stage of schistosomiasis on productivity and QOL of workers needs further research with a wide spectrum of workload in different industries. PMID- 12557940 TI - Serum levels of endothelin-1 (ET-1), interleukin-2 (IL-2) and amino-terminal propeptide type III procollagen (PIII NP) in patients with acute and chronic filariasis. AB - Filariasis, a mosquito-borne parasitic disease, is a worldwide health problem. There is still, some controversial concerning the diagnosis of acute and chronic infections. The serum levels of endothelin-1 (ET-1), interleukin-2 (IL-2) and amino-terminal propeptide Type III (PIII NP) was measured in patients with acute and chronic filariasis as compared with controls. The ET-1, IL-2 and PIII NP levels were significantly high in chronic cases than in acute. On the other hand, the serum levels of IL-2 and PIII NP were significantly high in acute cases than in the controls. These three immuno-mediators play role in the pathogenesis of filariasis particularly. The chronic cases. So, these mediators can be used as markers for diagnosis of human cases infected with chronic and acute filariasis. PMID- 12557941 TI - Studies on the present status of insecticides resistance on mosquitoes using the diagnostic dosages in El-Fayium Governorate, a spot area of malaria in Egypt. AB - Insecticides susceptibility tests were conducted using the World Health Organization diagnostic doses on the adult and larvae of Anopheles pharoensis and Culex pipiens mosquitoes from Fayium Governorate in Egypt. Insecticides used were malathion, fenitrothion, temephos, diazinon, bromophos and fenthion from organophosphorous group and deltamethrin, permethrin and cypermethrin from synthetic pyrethroid group while propoxur was from carbamate group. Larvae of A. pharoensis were resistant to fenitrothion and susceptible to other insecticides used, while the adults were susceptible to malathion, deltamethrin, fenitrothion and premethrin. Larvae of C. pipiens were resistant to temephos, fenitrothion, bromophos and fenthion and susceptible to malathion, permethrin and diazinon, while the adults were resistant to fenitrothion, permethrin and propoxur and susceptible to deltamethrin, cypermethrin and malathion. These results are important for the success of the control programmes and monitoring the susceptibility status of vectors. PMID- 12557942 TI - The response of Musca domestica (L.) in farms sprayed for long periods by organochlorine to synthetic pyrethroids. AB - Resistance ratios for Musca domestica collected from animal farms and poultry units, sprayed by organochlorine insecticides for long periods and standard laboratory strain were determined. The data indicated high significant resistance in animal farms population to organochlorine compounds and cross-resistance to synthetic pyrethroids. The organochlorine resistant and standard laboratory strains were examined for knockdown and rate of penetration to two synthetic pyrethroids. Standard laboratory strain was knocked down within few minutes by lower or higher dosages. Resistant strain was unaffected by lower dosages within 24 hours and partly by higher dosages. Penetration was faster in the resistant than in the standard strain. However, significant differences did not occur until after the standard laboratory females become moribund. The higher penetration rate in the resistant housefly adults was attributed to their continued activity throughout the exposure periods. PMID- 12557943 TI - Surface topography of Erilepturus hamati Manter, 1947 (Digenea, Family: Hemiuridae) by scanning electron microscopy. AB - The tegumental ultrastructure of adult Erilepturus hamati was studied by scanning electron microscopy (SEM). Adult worms were recovered from intestine of Red Sea fishes, Pherapon jarbua and Lethrinus mahsena collected at Hurghada, Red Sea, governorate. The study revealed the presence of three types of sensory papillae: type I button papillae around suckers, type II ciliated dome papillae scattered on tegumental surface of soma and type III domed papillae distributed on tegument of posterior extremity of soma. High magnification of tegumental surface of the pit in which the ecsoma is retracted showed that it has a number of small dermal outgrowths. Many pores are scattered on the tegumental surface. The tegumental ultrastructure of the present genus was compared with those of other genera of trematodes, previously described from the Red Sea fishes. PMID- 12557944 TI - Two types of scab mites infesting man and sheep in North Sinai. AB - Man and sheep were examined for scab mites infestations. Two types of mites were recovered, Sarcoptes scabiei and Psoroptes ovis. In sheep, 197 out of 939 (20.98%) were infested with S. scabiei (4.05%) and P. ovis (16.93%). The highly infested sheep were recorded in El Hassanah (28.22%) and the least infested ones were in Al Arish (15.85%). In man, 69 out of 790 (8.7%) were infested with S. scabiei (8.1%) and P. ovis (0.63%). The highly infested patients were recorded in El Hassanah (16.2%) followed by Rafah (11.7%), Bir Al-Abd (7.8%), and the least infested one was recorded in Al Arish (4.3%). In man, infestation was seen on one site in 15 patients (21.7%), on two sites in 32 patients (46.4%) and on more than two sites in 22 patients (31.9%). The highly infested site was the upper limbs followed by the lower limbs. Infestation on the face and ear, genital organs and abdomen were also encountered. Infestation on two sites was predominant on the upper and lower limbs followed by the lower limbs and genital organ. The results indicate that transmission of both scab mites, S. scabiei and P. ovis from sheep to man do occur. Infestation with scab mites was high in shepherds (79.7%) than in non shepherd patients (20.3%). However, man was more susceptible to S. scabiei than to P. ovis. PMID- 12557945 TI - Prevalence of external parasites in the south eastern desert of Egypt. AB - External parasites in the triangle region (Halaib & Shalatin) affecting the animal health were studied. Ectoparasites were collected in several sites by using bait traps and directly from animal bodies. Results indicated the presence of twelve species of insects belonging to seven genera included in three families (Calliphoridae, Muscidae and Sarcophagidae). Concerning ectoparasites on animal bodies, there were two species of biting lice infested goats and sheep (Bovicola caprae and B. ovis, respectively) and two species of sucking lice on goats (Linognathus africanus and L. stenopsis). Melophagus ovinus (family Hippoboscidae) collected from goats. Moreover, all camels suffered infestation with hard ticks four Hyalomma species. On the other hand, sheep and goats were infested with two Rhipicephalus species and one Haemaphysalis species. PMID- 12557946 TI - Cyclocoelum microstomum and C. obscurum (Trematoda: Cyclocoelidae) from air sacs of waterfowl in Egypt: early life history, morphological and histological studies of adult. AB - Two species of genus Cyclocoelum are recorded from air sacs of waterfowls, collected from Egypt. Fully morphological and histological descriptions for the adult stage of C. microstomum from moorhen Gallinula chloropus are presented. Also egg and miracidium stages are described. The second species, C. obscurum collected from coat Fulca sp. also described. Some problematic status of the genus Cyclocoelum are discussed include evaluation of some morphological and histological criteria also the migration and location of worm within the birds. PMID- 12557947 TI - The role of Fas system [Fas-FasL] as an inducer of apoptosis in cutaneous leishmaniasis: a combined human and experimental study. AB - Cutaneous leishmanial lesions as well as serum samples from both infected human and Swiss-albino mice were used to investigate the role of Fas system (Fas-FasL) as an inducer of apoptosis and other leishmanicidal cytokines in the disease development in cutaneous leishmaniasis. Soluble Fas was estimated by ELISA, other leishmanicidal cytokines were detected by PAP technique and tissue Fas by RT-PCR. Results showed a significant increase in interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha), nitric oxide (NO) and soluble Fas in both infected human and mice. As regards the tissue Fas, there was marked expression in human samples while in murine samples, the expression was reduced in chronic infected mice than in the late acute infected animals which can explain the progression of the lesion in the animals. This study confirms the role of Fas system as an inducer of apoptosis. PMID- 12557948 TI - Immunity to schistosomiasis before puberty: humeral or cell mediated. AB - To determine if the cell mediated immunity, induced by T-helper type-1 lymphocytes (Th1) response, during schistosomiasis mansoni has the potential to protect against infection, intensities of infections and re-infections, reflected in the egg count were followed up to 20 months among 119 individuals aged 5-22 years (Ys) with different number of previous infections whose yearly levels and pattern of water contact were similar. They were classified into 5 groups. Delayed hypersensitivity skin tests (DHT) to adult schistosome excretory secretary antigens (ESAgs) and anti-schistosomula (ESAgs) isotypes were measured on detecting re-infection. The group with a mean age of (8.6 +/- 2.6 Ys) and infected less than 5 times showed only 6.5 percentage reduction of the egg count (PREC) and low cellular and humoral responses. Th1-associated cellular (DHT) and antibody responses (IgG2, IgG3) to the five infections were significantly higher in the (13.5 +/- 1.4 Ys) than in (18 +/- 2.2 Ys) age group. This was reflected in significant difference in PREC; being 41.5% among the first and 13.5% among the second. Th2-associated antibody responses (IgG1, IgG4, IgE) went on rising as patients allowed for age and number of infections increased over 5, being significantly higher in the (19 +/- 1.8 Ys) than in (14 +/- 1.1 Ys) age groups with PREC 45.5% and 12.9% respectively. These results imply a substantial protective role for cell mediated immunity in the pre-puberty stage and provide evidence that Th1-based vaccination strategy can work if augmented. PMID- 12557949 TI - Evaluation of circulating Fasciola antigens in specific diagnosis of fascioliasis. AB - Ninety-eight individuals were included in this study, 60 of them were passing Fasciola eggs, 20 were infected with other parasites but not Fasciola (infected control group) and 18 individuals were free from parasite infection (normal control group). Circulating Fasciola antigens (CFAg) and anti-Fasciola IgG were estimated by ELISA technique among all groups. Highly significant increased level of CFAg was observed among Fasciola-infected cases when compared with normal control group. The percentage positivity of CFAg was 95% among Fasciola-infected cases. Solid specificity was observed by the test. Moreover, a highly significant increase in the antigen level was detected among moderately infected patients compared with lightly infected group. The level of CFAg was significantly higher in the hepatomegalic cases than those without organomegaly. Anti-Fasciola IgG antibodies were detected in only 35% of Fasciola infected cases, which indicates much lower sensitivity of the antibody detection than antigens. No correlation between the level of the circulating antibodies and either the intensity of infection or the clinical presentations was observed. PMID- 12557950 TI - Fine structural studies on the intestinal trematode, Gyliauchen volubilis Nagaty 1956 from fish Siganus luridus in the Red Sea, Egypt. AB - For the fine structure of Gyliauchen volubilis, sections of the finger-like projections showed the structure of a segmented body, which is not attached with the outer membrane of the tegument. Each projection consists of one to four segments, the smallest segment is triangular in shape and located on the top of the projection, enveloped by a triple layered membrane, with electron dense structures in the cytoplasm. Two types of vesicles are scattered in the tegumental matrix; vesicle with electron dense body, and multigranulated vesicle. Spherical fragment also contains electron dense structures and starts as a round projection of the tegument that detaches itself from the rest of the tegument. The presence of these fragments could be evidence of exocytosis for this trematoda. A layer of vacuoles measuring 1 to 1.5 microm long and 0.5 to 0.8 microm wide, are located beneath the matrix of the tegument which could be the douse of release product. For some areas of trematode, larger vacuoles measure 2.6 to 4.3 microm long and 0.5 to 0.8 microm wide, are found near the tegumental surface but in small number. There are two layers of subtegumental myofibers, circular and longitudinal. The circular muscle myofibers are grouped in bundles whereas the longitudinal myofibers are densely distributed adjacent to the circular fibers. Numerous polygonal structures whose function has not yet been determined are located beneath the muscle layers. PMID- 12557951 TI - Effects of the entomopathogenic nematode, Heterorhabditis bacteriophora HP 88 and azadirachtin on the immune defence response and prophenoloxidase of Parasarcophaga surcoufi larvae (Diptera: Sarcophagidae). AB - Each third larval instar (155-175 mg) of the fleshfly, Parasarcophaga surcoufi was injected with 20 and 40 infective juveniles (IJs) of entomopathogenic nematode, Heterorhabditis bacteriophora, and with 0.5 and 1 microg of azadirachtin. Injection with 20 IJs decreased the total haemocyte count (THC) up to 40 h post-injection, except for its increase at 20 h. On the contrary, injection with 40 IJs increased the THC during the hours post-injection, except for its decrease at 40 h. The injection of nematode (40 IJs) decreased, the percentage of differential haemocyte count of P. surcoufi larvae, specially the granulocytes and plasmatocytes at 40 h. Whereas, prohaemocytes and oenocytoids were increased at 40 h post-infection. The granulocytes and plasmatocytes were engaged with encapsulation of the nematode, H. bacteriophora. The released symbiotic bacterium, Xenorhabdus luminescens was poorly phagocytosed by the granulocytes. After injection of the larvae with azadirachtin, THC and the percentage of the number of oenocytoids were particularly increased after 40 h of injection with azadirachtin; plasmatocytes and granulocytes were decreased at 10 h post-injection with 0.5 microg and at 40 h post-injection with 1 microg of azadirachtin. The most prominent haemocyte deformities in P. surcoufi larvae treated with azadirachtin were the release of cytoplasmic components from granulocytes together with bulging or lysis of plasmatocytes. Maximal haemolymph phenoloxidase (PO) activity was obtained at pH 6.2 and 6.6. This activity increased with increasing the pre-incubation time using laminarin, alpha chymotrypsin and trypsin as activators. Injection of both nematode and azadirachtin significantly suppressed the haemolymph PO activity even in the presence of the activators laminarin, alpha-chymotrypsin and methanol. This suppression was dose-dependent and reached maximum at 30 h post-injection. The electrophoretic haemolymph protein profile was time-dependent as indicated by appearance and disappearance of protein bands. After 40 h post-injection with the nematode all the protein bands were replaced by new ones (probably containing immune protein). However, at this time injection with azadirachtin (1 microg) lead to appearance of 10 new protein bands & disappearance of an equal number of the present band. PMID- 12557952 TI - Immunodiagnosis of bovine fasciolosis using Fasciola hepatica excretory-secretory antigens ELISA. AB - Two hundreds blood samples and the corresponding faecal and/or bile (114 Egyptian buffaloes, 68 native breed cattle and 18 imported cattle) were examined for Fasciola infection. ELISA revealed that 8 (7.00%) buffaloes, 52 (76.50%) native breed cattle and 15 (83.33%) imported cattle were positive. Parasitologically, the results revealed that 38 (33.33%) buffaloes and 18 (26.50%) native breed cattle were positive, while all imported cattle were free from Fasciola. So, ELISA technique using F. hepatica excretory-secretory (E/S) antigens is of diagnostic value for cattle fasciolosis rather than for buffaloes. PMID- 12557953 TI - Bacteraemia in children. AB - OBJECTIVES: (i)To know the etiology of bacteraemia in children, (ii) To learn the antibiotic sensitivity pattern of the isolates. METHOD: Over the period of thirteen months 4,368 blood samples (for blood culture) were collected from children in the age group of 0 day-14 years, suspected of having fever and sepsis. Blood samples were collected for blood culture from each case. Organisms were isolated and identified by conventional methods. Antibiotic susceptibility for each isolate was determined by using modified Stokes method. RESULT: 1,001 cases (22.9%) were culture positive. Incidence of bacteraemia in neonates was 521(33.94%). Gram negative organisms were the most predominant isolates (88.8%). Commonest was Klebsiella 471 (47.1%) followed by Salmonella sp. 162 (16.2%) and Pseudomonas 80 (8%) whereas in gram positive, Staphylococcus aureus 76 (7.6%) was the most common. Maximum sensitivity was seen by sulbactum/cefaperazone combination-969 (98.2%) by all isolates. Linezolid 97 (99.0%) was the most sensitive drug for gram positive isolates. CONCLUSION: Gram negative multidrug resistant organisms were the main cause of septicemia in all the age groups. Therefore great caution is required in selection of antibiotic therapy. PMID- 12557954 TI - Measles in a hilly hamlet of northern India. AB - OBJECTIVE: To ascertain the epidemiological links and risk factors responsible for the epidemic of measles in the village Astani in Himachal Pradesh. METHODS: All the children less than twelve years who were present on 27th July 1997 were evaluated. A questionnaire requesting data on vaccination history and symptoms of measles was administered and complete physical examination including anthropometry was recorded in each child by the authors. The data also included complications (including that of death) secondary to measles infection. RESULTS: A total of 48 children less than 12 years, present in the hamlet on 27th July 1997, were examined, out of which 28 (58%) were affected and 20 (42%) were unaffected. The immunization coverage in the affected children was 33% in contrast to 70% coverage in the unaffected group. Vaccine efficacy in the present epidemic was 51%. The complication rate was 59%, which included one death. The anthropometric data showed that 92% of the affected children were malnourished. All the children were given age appropriate dose of Vitamin A and children of the nearby villages were vaccinated against measles as a part of outbreak control. CONCLUSION: This study clearly highlights the need to achieve and sustain high immunization coverage along with strengthening of the routine surveillance systems in remote village of India. PMID- 12557955 TI - Impact assessment of mass measles vaccination. AB - OBJECTIVE: The mass measles vaccination campaign was conducted in the slums of Surat City, in Gujarat State, as a part of urban measles control initiative in India. One dose each of the vaccine was administered to children in the age range of 9-59 months residing in these slums, regardless of their previous vaccination status. METHODS: One year later, (October 2000), the present study was carried out in order to assess the impact of the mass vaccination campaign on the vaccination coverage and on the incidence of measles by comparing the findings with those of the baseline survey carried out in May 98. This was a retrospective study with a recall period of the preceding year. 3,147 children under five were studied in thirty slum clusters selected by the cluster sampling method. The parents/caretakers of these children were interviewed for information on any episode of fever with rash conforming to the case definition. RESULT: The incidence rate for measles declined from 7.7 percent reported in the baseline (May 1998) to 3.5 percent in the impact assessment study. The incidence was 8 times higher in unvaccinated children. The mean and median age at contracting the illness increased from 26 +/- 14.2 months and 26 months in the baseline to 30.9 +/- 14.7 months and 30 months respectively in the impact assessment. The vaccination coverage had improved from 48.3 percent to 73.7 percent following the campaign. CONCLUSION: The compaign increased vaccination coverage decreased disease incidence and caused a shift towards higher age-groups in vaccinated children. PMID- 12557956 TI - Physical status of newborns and neonatal outcome. AB - OBJECTIVE: To determine the extent to which physical status at birth is associated with neonatal mortality and the causes of mortality vis-a-vis size at birth and gestational age. METHOD: 11,223 consecutive live births completing 26 weeks of gestation and weighing > or = 500 gm were included in the study. Birth weight and chest circumference were recorded as per WHO guidelines. Gestational age was calculated on the basis of L.M.P. and the new Ballard's score. Deaths occurring in the hospital within 28 days were recorded. Percentile values of gestational age specific birth weights were calculated separately for singletons and multiple births. Percentage of SGA was calculated with reference to WHO recommended values. Birth weight-gestational age-specific mortality rates were calculated at 2 wk and 500 gm intervals. RESULT: Low-birth-weight babies constituted 39.8% of the total, much in excess of WHO recommended figure of 15%. 76% deaths occurred among LBW babies and 56.2% among preterms. Mortality showed remarkable decline as the birth weight increased to 2,000 gm. The lowest mortality was among singletons weighing 2,500-3,000 gm and of 38-40 weeks gestation. Prevalence of SGA at 40 and 42 weeks were 73.7% and 83.6% respectively. But, if SGA babies not categorised as LBW were excluded, the values came down to 32% and 36% respectively. 36% of all deaths occurred during the first 24 hrs of birth; asphyxia and related causes contributing to 50% of it. CONCLUSION: Cut-off value of 2,000 gm instead of 2,500 gm for birth weight may be preferable in countries where most LBW babies are SGAs. Simultaneously, deaths in non-LBW babies due to perinatal causes contribute sgnificantly to total neonatal mortality and need due attention through sensitising obstetricians in essential newbom care and timely Intervention. PMID- 12557957 TI - Clinico-pathological profile of pediatric lymphadenopathy. AB - OBJECTIVE: To study the clinico-pathological profile of significant pediatric peripheral lymphadenopathy and to arrrive at an etiological diagnosis. METHODS: Prospective study in a tertiary care hospital setting. One hundred consecutive children reporting to pediatric OPD from 1 January 1995 to 31 December 1998, aged 1 month to 12 years were studied. RESULTS: The commonest aetiology diagnosed was tubercular lymphadenitis in 35% cases, followed by chronic tonsillopharyngitis in 15% cases. Lymphomas, AIDS and infectious mononucleosis constituted 3, 2 and 1 cases each. Aetiology could not be ascertained in 44 (44%) children even after detailed haematological, microbiological, radiological and serological investigations. FNAC's sensitivity and specificity as compared to 'gold standard of excision lymph node biopsy was 94% and 100% respectively. CONCLUSION: Tubercular lymphadenitis was the commonest treatable entity of significant pediatric peripheral lymphadenopathy. A majority of the cases even after thorough evaluation, remained undiagnosed. FNAC as a diagnostic modality is almost as sensitive and as specific as excision lymph node biopsy when an adequate aspirate is examined by expert eyes. PMID- 12557958 TI - Nephrotic syndrome and proteinuria. PMID- 12557959 TI - Isolated asymptomatic proteinuria. AB - Proteinuria is occasionally detected on "routine" examination (not done for a suspected renal or urinary tract problem) and often presents a diagnostic problem. Even mild proteinuria is abnormal and needs an explanation. Its degree and persistence should be ascertained and associated hematuria looked for. Nonpathological (including orthostatic) proteinuria and renal tubular disorders should be excluded. Persistent proteinuria is mostly due to a primary or secondary (eg hepatitis B) glomerular condition, which requires expert evaluation of renal biopsy. Aggressive therapy may be needed in some cases. The intensity of proteinuria can be decreased with ACE inhibitors. Long-term observation is essential. PMID- 12557961 TI - Steroid resistant nephrotic syndrome. AB - Steroid Resistant-Nephrotic Syndrome (NS) is a chronic, progressive disorder affecting upto 10% of all children with NS. It causes morbidity and mortality due to persistent edema, hypertension, hyperlipidemia, thrombosis and infection. Progression to renal failure was thought to be inevitable in survivors. Recent insights into the pathogenesis of the disease has identified several responsible genes and proteins. Studies have shown that long term aggressive therapy with combinations of steroids, alkylating agents and cyclosporine, cause complete or partial remission in 20-80% patients. The use of nonspecific renal protective agents such as the angiotensin converting enzyme inhibitors, angiotensin 2 receptor blockers, and anti-lipid agents retard disease progression. Although these are indications of significant improvement in outcome, further multicentre controlled studies are required to determine the optimum drugs and regimens to be used. PMID- 12557960 TI - Steroid sensitive nephrotic syndrome. AB - Nephrotic syndrome in children is a common recurrent disease. Most of the cases are due to minimal change disease with a favourable outcome. More than 90% of children with minimal change disease respond to corticosteroid therapy (steroid sensitive nephrotic syndrome). 40-60% experience frequent relapses or have steroid dependence. These children require frequent corticosteroid therapy and/or immunomodulators or treatment with immunosuppressants, and are at high risk of cumulative steroid toxicity and side effects of cytotoxic therapy. Children with frequent relapses or steroid dependence should be managed in consultation with a pediatric nephrologist. Despite relapsing course, progression of minimal change nephrotic syndrome to end stage renal disease is extremely rare. PMID- 12557962 TI - Acute and crescentic glomerulonephritis. AB - Acute nephritic syndrome is clinically characterized by hematuria, proteinuria, oliguria, and volume overload with or without azotemia and histologically be acute proliferative glomerulonephritis. Acute post streptococcal glomerulonephritis is the commonest cause in children. There is a preceding infection prior to this condition in majority. This is one of the comonest causes of renal edema in children. Early recognition, prompt and aggressive therapy and adequate follow-up are mandatory. Prognosis is usually good unless associated with severe renal failure and crescentic glomerulonephritis where the outcome is relatively poor unless treatment is early and adequate. Pathologically acute proliferative nephritis is with diffuse proliferative glomerulonephritis with or without crescents. Immunosuppressive therapy is not needed in simple acute proliferative glomerulonephritis but is essential in modifying the outcome of crescentic glomerulonephritis. Delayed resolution, severe renal failure at onset, progressive renal failure and associated systemic features like skin rashes, joint pains, hepatosplenomegaly and persistent fever are the indications for biopsy. Overall the prognosis in classical post streptococcal acute proliferative glomerulonephritis is good. PMID- 12557963 TI - Hypertension in children. AB - Abstract. Hypertension is a major public health issue in industrialized nations, affecting approximately 20% of adults. Hypertension often begins in childhood, and children with hypertension are very likely to be seen with this disorder as adults. The close associations of hypertension with atherosclerosis, coronary and cerebrovascular disease, diabetes and end-stage renal disease make it a major contributor to the most common causes of morbidity and mortality in adult population. (1) According to many studies performed in adult populations, the effective treatment of hypertension reduces the risk of coronary heart disease, stroke, renal disease and congestive heart failure. (2) PMID- 12557964 TI - Terminal myelocystocele. AB - Terminal myelocystocele is an unusual form of occult spinal dysraphism. It consists of a cystic dilatation of a low-lying terminal cord herniated posteriorly through a skin covered lumbosacral spina bifida. This condition is often associated with OEIS complex i.e. opmphalocele, exstrophy of the bladder, imperforate anus and spinal abnormality. We studied 4 cases of terminal myelocystocele. They revealed no preoperative neurological deficit. None of these had associated OEIS complex. One of the cyst was unique due to presence of copious amount of pus in its cavity. All four cases underwent successful repair and surgery remained uneventful in all of them. No child showed neurological deterioration during the follow-up. PMID- 12557965 TI - Neonatal diabetes with hyperchylomicronemia. AB - Neonatal diabetes mellitus (NDM) is defined as hyperglycemia occurring in the first few weeks of life. It can be either transient (TNDM) or permanent (PNDM). A 25 days old newborn was brought to the hospital with restlessness, respiratory depression and cyanosis. He was born at term with a birth weight of 2,000 g. There was no consanguinity between his parents. His physical examination findings were as follows: Weight and height were under 3th percentile, he was hypoactive and dehydrated. Serum glucose level was 800 mg/dl; C-peptide was 0.41 ng/ml. Upon investigation for dyslipidemia in association with his neonatal diabetes, hyperchylomicronemia was found both in the patient and his father. Pancreatitis, anemia and cholestasis were also observed. Insulin treatment was started for his diabetes together with a special diet for dyslipidemia. At the end of 28 months of follow-up, dyslipidemia has resolved but the need for insulin therapy was still existing. However, TNDM was considered in differential diagnosis because he was small for gestational age (SGA) at birth and his symptoms had started at the 25th day of the neonatal period. Delayed recovery from insulin dependency brought out the possibility of PNDM. Furthermore, neonatal diabetes combined with hypechylomicronemia is a rare clinical picture. Reported cases of NDM with different clinical evaluation will help to better understanding of this disorder. PMID- 12557966 TI - Medullary sponge kidney. AB - Medullary sponge kidney is a benign asymptomatic developmental anomaly of the kidney mostly seen in adult females. Presentation in childhood is uncommon. Urinary tract infection, nephrolithiasis, hematuria and hypercalciuria are the common complications. We report a eleven-year-old female child who presented with recurrent urinary tract infection and nephrolithiasis and was found to have bilateral medullary sponge kidney. PMID- 12557967 TI - Sirenomelia dipus in a dizygotic twin. AB - A rare case of sirenomelia in a LBW, IUGR & second of a dizygotic twin having multiple congenital anomalies is being reported. Apart from the features of sirenomelia, the newborn had a single umbilical artery, radial hypoplasia, total anorectal & urogenital agenesis and undetermined sex. The single umbilical artery had a normal origin from the iliac artery. There was no Vitelline arterial steal phenomenon. Theories put forward regarding etiopathogenesis are discussed. We conclude that the anomalies in the present case are difficult to be explained with earlier proposed theories. This puts an emphasis on reconsideration of old theory of genetic injury to the embryo by unknown factors. PMID- 12557968 TI - Baller-Gerold syndrome. AB - Craniosynostosis is known to be associated with a large number of inherited disorders of childhood. Its presence along with absent thumbs and ectopic anus is the characteristic feature of one such extremely rare disorder, described as the Baller Gerold syndrome. The typical features are being reported here. PMID- 12557969 TI - Posterior fossa oligodendroglioma. AB - Oligodendroglioma are the tumors of glial cells. They are rare in children and are more common in the cerebral hemispheres. A rare case of infratentorial oligodendroglioma in a female child is being reported here. PMID- 12557970 TI - Are we teaching some wrong facts pertaining to oral polio vaccine? PMID- 12557971 TI - Accuracy of CT-based thickness measurement of thin structures: modeling of limited spatial resolution in all three dimensions. AB - Measurement of the width of thin structures such as the cortical shell of the vertebral body or femoral neck with computed tomography (CT) is limited by the spatial resolution of the CT system. Limited spatial resolution exists both within the CT image plane and perpendicular to it and can be described by the in plane point spread function (PSF) and the across-plane slice sensitivity profile (SSP), respectively. The goal of this study was to confirm that errors of thickness measurement of thin structures critically depend on the spatial positioning of the object and the spatial resolution limitations of CT in all three dimensions, and to assess the size of the errors themselves. We compared computer models that incorporated both effects to experimentally assessed cortical thicknesses of the European Spine Phantom. Analysis included varying CT slice width, the orientation of measurement and angle beta of misalignment of longitudinal scanner and phantom axes. Agreement of models with measurements was good in all configurations with an overall error of 0.17 mm. This showed that PSF and SSP are adequate system characteristics to predict deviation of measured values from true widths. Errors between measurements and true cortical thickness values delta(true) averaged to 1.5 mm were strongly positively correlated with slice width d and beta. When the across-plane partial volume effect was eliminated, limited in-plane resolution still accounted for overestimation of delta(true) by 0.68 (137%), 0.27 (27%), and 0.06 mm (4%) for delta(true)=0.5, 1.0, and 1.5 mm, respectively. For delta(true) of 1.0 mm and above, it was shown that although the absolute cortical thickness values might not be accurately measurable, relative differences between two values are reflected in measurement. Implications for cortical thickness measurement are that the spinal cortical shell is too thin, whereas accurate assessment at locations of the femoral neck exhibiting a thicker cortical shell of both difference and absolute values should be possible with CT even for larger misalignment angles, especially when a smaller CT slice width is chosen. PMID- 12557972 TI - Monte Carlo dosimetry of a new 192Ir pulsed dose rate brachytherapy source. AB - A new microSelectron pulsed dose rate source has been designed, containing two active pellets instead of one inactive and one active pellet contained in the old design, to facilitate the incorporation of higher activity up to 74 GBq (2 Ci). In this work, Monte Carlo simulation is used to derive full dosimetric data following the AAPM TG-43 formalism, as well as the dose rate per unit air kerma strength data in Cartesian, "away and along" coordinates for both source designs. The calculated dose rate constant of the new PDR source design was found equal to lambda=(1.121 +/- 0.006) cGy h(-1) U(-1) compared to lambda = (1.124 +/- 0.006) cGy h(-1) U(-1) for the old design. Radial dose functions of the two sources calculated using the point source approximated geometry factors were found in close agreement (within 1%) except for radial distances under 2 mm. At polar angles close to the longitudinal source axis at the sources' distal end, the new design presents increased anisotropy (up to 10%) compared to the old one due to its longer active core. At polar angles close to the longitudinal source axis at the sources' drive wire end however, the old design presents increased anisotropy (up to 18%) due to attenuation of emitted photons through the inactive Ir pellet. These differences, also present in "away and along" dose rate results, necessitate the replacement of treatment planning input data for the new microSelectron pulsed dose rate source. PMID- 12557973 TI - Spatial resolution of magnetic resonance imaging Fricke-gel dosimetry is improved with a honeycomb phantom. AB - The spatial accuracy of magnetic resonance imaging (MRI) Fricke-gel dosimetry is limited by diffusion of ferric ions. This paper describes a honeycomb structure to limit diffusion of Fe3+ ions in a three-dimensional phantom. Such a phantom containing the dosimeter gel was irradiated to a known dose distribution. Maps of dose distributions were produced from the MR images acquired at 2 and 24 hours after the dose was given. The dose distribution maps verified that the honeycomb structure precludes ion diffusion from one honeycomb cell to another, thus improving the usefulness of MRI Fricke-gel dosimetry. PMID- 12557974 TI - Validation of a pencil beam model-based treatment planning system for fast neutron therapy. AB - Treatment planning systems (TPSs) are used to compute dose delivered to the patient. In the case of fast neutron therapy, TPSs are mostly not of general purpose but are dedicated to one facility. This is due to the few fast neutron facilities worldwide and due to the high variation in the neutron energy distributions. Efforts have been undertaken to develop a new TPS that could be applied to all the existing fast neutron facilities. The University Hospital of Essen operates a d (14 MeV) + Be fast neutron beam and the TPS used is based on an empirical model. In a previous study, the empirical model has been evolved to a pencil beam model of 35 monoenergetic neutron beams. Monte Carlo techniques have been utilized to compute distributions of the energy deposition due to primary and scattered neutrons in a simple geometry water phantom. The experimental validation of the method is now presented. Depth dose curves in water of monoenergetic neutrons have been derived from the distributions of energy deposition. The resultant depth dose curves have been utilized in order to determine the depth dose curves of the fast neutron beam of the Essen facility for the 14 radiation field sizes available in this facility. This determination requires the initial neutron spectrum. As this spectrum could not be measured at the Essen facility, the initial neutron spectrum of the Physikalisch Technische Bundesanstalt, Braunschweig, Germany, which operates the same cyclotron, was used. The calculated depth dose curves were compared to experimental depth dose curves that have been obtained in water at the University Hospital of Essen. The comparison between calculated and experimental depth dose curves showed significant deviations in the case of large radiation fields and of depth less than 5 cm. In the case of radiation field areas less than 150 cm2 and depth more than 5 cm (usual clinical situation), the measured and calculated values are in a good agreement. In the case of clinical situation, the dependence on the radiation field size is relatively well taken into account by the model presented here. PMID- 12557975 TI - Monte Carlo dose characterization of a new 90Sr/90Y source with balloon for intravascular brachytherapy. AB - Beta emitting source wires or seeds have been adopted in clinical practice of intravascular brachytherapy for coronary vessels. Due to the limitation of penetration depth, this type of source is normally not applicable to treat vessels with large diameter, e.g., peripheral vessel. In the effort to extend application of its beta source for peripheral vessels, Novoste has recently developed a new catheter-based system, the Corona 90Sr/90Y system. It is a source train of 6 cm length and is jacketed by a balloon. The existence of the balloon increases the penetration of the beta particles and maintains the source within a location away from the vessel wall. Using the EGSnrc Monte Carlo system, we have calculated the two-dimensional (2-D) dose rate distribution of the Corona system in water for a balloon diameter of 5 mm. The dose rates on the transverse axis obtained in this study are in good agreement with calibration results of the National Institute of Standards and Technology for the same system for balloon diameters of 5 and 8 mm. Features of the 2-D dose field were studied in detail. The dose parameters based on AAPM TG-60 protocol were derived. For a balloon diameter of 5 mm, the dose rate at the reference point (defined as r0 = 4.5 mm, 2 mm from the balloon surface) is found to be 0.01028 Gy min(-1) mCi(-1). A new formalism for a better characterization of this long source is presented. Calculations were also performed for other balloon diameters. The dosimetry for this source is compared with a 192Ir source, commonly used for peripheral arteries. In conclusion, we have performed a detailed dosimetric characterization for a new beta source for peripheral vessels. Our study shows that, from dosimetric point of view, the Corona system can be used for the treatment of an artery with a large diameter, e.g., peripheral vessel. PMID- 12557976 TI - Evaluation of external beam radiotherapy and brachytherapy for localized prostate cancer using equivalent uniform dose. AB - Various radiotherapy (RT) modalities, such as external beam radiotherapy (EBRT) and permanent/high-dose-rate (HDR) brachytherapy, have been used for the management of localized prostate cancer. Using the linear-quadratic (LQ) model, we compared the relative merits of these modalities in terms of equivalent uniform dose (EUD) and tumor control probability (TCP). The LQ parameters (alpha = 0.15 Gy(-1) and alpha/beta = 3.1 Gy) determined recently from compiled clinical data, as well as other sets of LQ parameters for prostate cancer, were used to carry out the EUD and TCP calculations. A computer code was developed for this purpose. We calculate the EUD for some common RT modalities, and present the corresponding TCP data predicted for a sample patient group (high-risk). Biological equivalence of treatment outcome among various RT modalities is demonstrated. The model suggests that the hypofractionation is preferred in terms of tumor control, due to the lower alpha/beta ratio. Also, the current combined treatment schemes (initial EBRT + permanent/HDR brachytherapy boost) provide higher EUD and TCP than these monotherapies. The study shows that EUD is less sensitive to model parameters than TCP, and EUD can be used to compare and to optimize treatment plans involving different RT modalities. Techniques to further optimize and/or to combine external beams with brachytherapy for better treatment outcomes are proposed. PMID- 12557977 TI - Emission spectra of gadolinium-158. AB - The photon and electron spectra of 158*Gd as result of the (n, gamma)-reaction on 157Gd were calculated with IMRDEC Monte Carlo computer program. The relaxation probabilities were taken from EADL file of the Lawrence Livermore National Laboratory. Because this file does not consist of data for some N and O transitions, they were additionally determined by Chen applying the DHS j-j scheme. Two schemes are applied to calculate the energies of the transitions: (1) a simple (Z + 1)/Z scheme, and (2) accurate calculation solving the relativistic Dirac equations. PMID- 12557978 TI - IVBTMC, a Monte Carlo dose calculation tool for intravascular brachytherapy. AB - A new Monte Carlo code (IVBTMC) is developed for accurate dose calculations in intravascular brachytherapy (IVBT). IVBTMC calculates the dose distribution of a brachytherapy source with arbitrary size and curvature in a general three dimensional heterogeneous medium. Both beta and gamma sources are considered. IVBTMC is based on a modified version of the EGSNRC code. A voxel-based geometry is used to describe the target medium incorporating heterogeneities with arbitrary composition and shape. The source term is modeled using appropriate phase-space data. The phase-space data are calculated for three widely used sources (32P, 90Sr/90Y, and 192Ir). To speed up dose calculations for gamma sources, a special version of IVBTMC based on the kerma approximation is developed. The accuracy of the phase-space data model is verified and IVBTMC is validated against other Monte Carlo codes and against reported measurements using radio-chromic films. To illustrate the IVBTMC capabilities, a variety of examples are treated. 32P, 90Sr/90Y, and 192Ir sources with different lengths and degrees of curvature are considered. Calcified plaques with regular and irregular shapes are modeled. The dose distributions are calculated with a spatial resolution ranging between 0.1 and 0.5 mm. They are presented in terms of isodose contour plots. The dosimetric effects of the source curvature and/or the presence of calcified plaques are discussed. In conclusion, IVBTMC has the capability to perform high-precision IVBT dose calculations taking into account the realistic configurations of both the source and the target medium. PMID- 12557979 TI - Computer-assisted detection of colonic polyps with CT colonography using neural networks and binary classification trees. AB - Detection of colonic polyps in CT colonography is problematic due to complexities of polyp shape and the surface of the normal colon. Published results indicate the feasibility of computer-aided detection of polyps but better classifiers are needed to improve specificity. In this paper we compare the classification results of two approaches: neural networks and recursive binary trees. As our starting point we collect surface geometry information from three-dimensional reconstruction of the colon, followed by a filter based on selected variables such as region density, Gaussian and average curvature and sphericity. The filter returns sites that are candidate polyps, based on earlier work using detection thresholds, to which the neural nets or the binary trees are applied. A data set of 39 polyps from 3 to 25 mm in size was used in our investigation. For both neural net and binary trees we use tenfold cross-validation to better estimate the true error rates. The backpropagation neural net with one hidden layer trained with Levenberg-Marquardt algorithm achieved the best results: sensitivity 90% and specificity 95% with 16 false positives per study. PMID- 12557980 TI - An optimization algorithm for intensity modulated radiotherapy--the simulated dynamics with dose-volume constraints. AB - We have developed a new method for optimization in intensity modulated radiation therapy (IMRT) that makes use of simulated dynamics in a classical system of interacting particles. An analogy is drawn between intensity profile optimization in IMRT and relaxation to the equilibrium configuration in a dynamic system. The intensities of beamlets are equivalent to the positions of the virtual particles. The potential energy of the system is defined by the objective function, which determines the equations of motion for the virtual particles. In this paper, we present the implementation of dose constraints and dose-volume constraints. Our strategy is to optimize the dose to the planned target volume (PTV) while keeping all constraints to the organs at risk (OARs) satisfied rigorously. A simple quadratic objective function is used that only includes terms for PTV voxels. By this approach, no additional parameters other than that for prescribing desired dose and constraints, such as importance factors, are needed. The hard constraints that require non-negative beamlet intensities and that the dose at any voxel in an OAR cannot exceed a maximum tolerance, are implemented as semi transmittable potential barriers of infinite height. Dose-volume constraints are handled by placing hard constraints on partial volumes. Handling of the clinically applied constraints was tested using phantoms and clinical cases. Our results show that the dose-volume histogram (DVH) type of plan prescription can be fulfilled with satisfactory PTV coverage. In addition to the convenience of implementation, our method can achieve a high computational efficiency with the understanding of the dynamic behavior of the system. PMID- 12557981 TI - The impact of increased Al filtration on x-ray tube loading and image quality in diagnostic radiology. AB - Previous work has shown that for nine common radiographic projections (AP abdomen, AP cervical spine, LAT cervical spine, PA chest, LAT chest, AP hip, AP lumbar spine, LAT lumber spine, and AP pelvis) increasing the total x-ray tube filtration from 2.5 mm Al equivalent (the regulatory minimum for general diagnostic radiology) to 4.0 mm Al equivalent, reduces the average effective dose and average skin entrance dose by 9% and 16%, respectively, using a 400 speed screen-film system. In this study, the effects of this filtration increase on x ray tube loading and image quality were assessed. For the above projections and filtration increase, mean absolute and percentage increases in tube loading were 2.9 mAs and 15%, respectively, for a constant film density and fixed kVp. Tube current (mA) increases of 25% (a worst case) resulted in no statistically significant loss in focal spot resolution due to blooming for both large (1.2 mm) and small (0.6 mm) focal spot sizes, except at high mA low kVp techniques. The latter losses were below 10%, and when the image receptor blur was incorporated, the total system spatial resolution losses were on the order of one-quarter to one-half these values for typical clinical geometries. Radiographs of a contrast phantom taken with 2.5 and 4.0 mm total Al equivalent x-ray tube filtration were compared at 60, 70, 81, 90, 102, and 121 kVp. No statistically significant changes were observed with regard to (1) test object conspicuity as reported by three observers, (2) image contrast, as measured using a densitometer with a 3 mm aperture (+/-0.0017 OD, 95% confidence level), and (3) pixel value image noise, image contrast-to-noise ratios, and image signal-to-noise ratios, as measured using a scanning densitometer with a 12-bit acquisition depth and 85 micron pixel size (+/-2.5%, +/-3.1%, and +/-2.5%, 95% confidence levels, respectively). These results, combined with the linear no-threshold model for radiation risk and the ALARA principle, suggest that general radiography should be carried out using a minimum of 4.0 mm total Al equivalent filtration. PMID- 12557982 TI - Inductively coupled transmission of neuro-active signals: analysis of optimal parameters. AB - Fifteen parameters that play a role in the optimal transmission of therapeutic signals by inductively coupled implantable neurostimulator have been investigated. For this purpose, at first, a model of the system was constructed from which the system transfer function was obtained. Then, the relationship between the transfer gain and each parameter was evaluated using mathematical equations and a specifically built computer program. This study showed that the gain could be increased selecting small values for some parameters (the number of active coil windings, first radii of inner and outer paths of the core, heights of the core base and windings, position under the skin, internal resistances of the active and passive coils, tissue impedance between the contacts of electrode), and high values for the others (the number of passive coil windings, second radii of inner and outer paths of the core, frequency of the signal, relative magnetic permeability of the core). Critical saturation values were another considerable point. The nearest commercially available standard values should be preferred in practical applications. PMID- 12557983 TI - Respiration-correlated spiral CT: a method of measuring respiratory-induced anatomic motion for radiation treatment planning. AB - We describe a method for generating CT images at multiple respiratory phases with a single spiral CT scan, referred to as respiratory-correlated spiral CT (RCCT). RCCT relies on a respiration wave form supplied by an external patient monitor. During acquisition this wave form is recorded along with the initiation time of the CT scan, so as to "time stamp" each reconstructed slice with the phase of the respiratory cycle. By selecting the appropriate slices, a full CT image set is generated at several phases, typically 7-11 per cycle. The CT parameters are chosen to optimize the temporal resolution while minimizing the spatial gap between slices at successive respiratory cycles. Using a pitch of 0.5, a gantry rotation period of 1.5 s, and a 180 degrees reconstruction algorithm results in approximately 5 mm slice spacing at a given phase for typical respiration periods, and a respiratory motion within each slice that is acceptably small, particularly near end expiration or end inspiration where gated radiotherapy is to occur. We have performed validation measurements on a phantom with a moving sphere designed to simulate respiration-induced tumor motion. RCCT scans of the phantom at respiratory periods of 4, 5, and 6 s show good agreement of the sphere's motion with that observed under fluoroscopic imaging. The positional deviations in the sphere's centroid between RCCT and fluoroscopy are 1.1+/-0.9 mm in the transaxial direction (average over all scans at all phases +/-1 s.d.) and 1.2+/-1.0 mm in the longitudinal direction. Reconstructed volumes match those expected on the basis of stationary-phantom scans to within 5% in all cases. The surface distortions of the reconstructed sphere, as quantified by deviations from a mathematical reference sphere, are similar to those from a stationary phantom scan and are correlated with the speed of the phantom. A RCCT scan of the phantom undergoing irregular motion, demonstrates that successful reconstruction can be achieved even with irregular respiration. Limitations from x-ray tube heating in our current CT unit restrict the length of the scan region to 9 cm for the RCCT settings used, though this will not be a limitation for a multislice scanner. RCCT offers an alternative to the current method of respiration-triggered axial scans. Multiple phases of respiration are imaged with RCCT in approximately the same scanning time required to image a single phase with a triggered axial scan. RCCT scans can be used in connection with respiratory-gated treatment to identify the patient-specific phase of minimum tumor motion, determine residual tumor motion within the gate interval, and compare treatment plans at different phases. PMID- 12557984 TI - The provision of consultative physics services as commodities undermines the professionalism of medical physics. For the proposition. PMID- 12557985 TI - The provision of consultative physics services as commodities undermines the professionalism of medical physics. Against the proposition. PMID- 12557986 TI - Improve your marketing effectiveness and net income through better prospecting. AB - Prospecting is the process of finding customers who are ready to buy and can generate high net income for an organization. Leads for prospects come from three categories of sources: (1) organization-initiated; (2) acquired leads; and (3) marketing activity-initiated leads. Findings from a study of academic medical organizations showed a modest use of effective prospecting by hospitals surveyed and that there are opportunities to increase database marketing efforts. The data suggests that prospecting and its companion concept of qualifying are not fully integrated into many healthcare organization's marketing strategies and tactics. PMID- 12557987 TI - Assessing Medicaid patients' perceptions of the OB/GYN-patient relationship. AB - OBJECTIVES: The objective of this study was to understand how the dynamics of the health care provider-patient relationship differ between Medicaid patients and private pay patients in the context of obstetric care. Various aspects of the patient-physician relationship were examined including trust, commitment, dependence, social content, service quality, and behavioral outcomes such as satisfaction, referral behavior, ease of voice, and retention. METHODS: Questionnaires were mailed to a sample of mothers who had recently given birth. MANOVA was used to compare the means of Medicaid patients with private pay patients for the variables of interest in the study. RESULTS: Medicaid patients had lower commitment to their primary physician. They trusted the practice, the primary physician, and the other physicians in the practice less. They perceived themselves as less similar to both the overall practice and their primary physician and also rated their health care service experience lower. They were less satisfied and less likely to use the same practice for future pregnancies or make referrals. They also felt less comfortable voicing complaints. CONCLUSIONS: The evidence clearly indicates that Medicaid obstetric patients perceived their service experience more negatively than private pay patients. Health care providers know they must provide clinical quality for their patients, however, in treating Medicaid patients they need to focus on patient driven-quality as well. The results indicate that health care providers, particularly OB/GYNs, need to do a better job of determining and delivering the key performance criteria that Medicaid patients use to make trust judgements. PMID- 12557988 TI - Keeping the medical practice financially viable: a marketer's approach. AB - This paper explores a way that medical practices might develop a higher margin revenue source by offering a differentiated service that caters to patients who value more personalized service. Using SERVQUAL, German patients were surveyed to determine if there is a relationship between a desire for personalized service and a willingness to pay a premium for medical care. The results indicate that a greater desire for personalized care distinguishes those who pay a premium for medical care from those who do not. This suggests that medical practices can differentiate their service based on more personalized care and charge a higher fee for this service. PMID- 12557989 TI - Changing drivers' intentions and behaviours using fear-based driver fatigue advertisements. AB - This paper examines the effects of fear arousal and perceived efficacy on the acceptance and rejection of road safety advertising messages that are typical in Australia and New Zealand. Our results suggest that the level of fear arousal could be lowered without a significant effect on the message acceptance rates but could result in a lower rate of message rejection. Our results also suggest that the inclusion of explicit coping strategies in the road safety advertisements has a significant positive effect on message acceptance. PMID- 12557990 TI - Consumption value theory and the marketing of public health: an effective formative research tool. AB - Contemporary public health requires the support and participation of its constituency. This study assesses the capacity of consumption value theory to identify the basis of this support. A telephone survey design used simple random sampling of adult residents of Cherokee County, Oklahoma. Factor analysis and stepwise discriminant analysis was used to identify and classify personal and societal level support variables. Most residents base societal level support on epistemic values. Direct services clientele base their support on positive emotional values derived from personal contact and attractive programs. Residents are curious about public health and want to know more about the health department. Where marketing the effectiveness of public health programs would yield relatively little support, marketing health promotion activities may attract public opposition. This formative research tool suggests a marketing strategy for public health practitioners. PMID- 12557991 TI - Social Security and SSI as safety nets for the elderly poor. AB - Partial privatization of Social Security is being considered as an integral part of the future Social Security program for American retirees. Because privatization creates uncertainty about the amount of retirement income that future retirees may expect to receive, the issue of a safety net is critical. This article presents the findings from an empirical study that investigated the degree to which the current Social Security and Supplemental Security Income (SSI) programs provide a safety net, separately and in combination, to the elderly poor. The major findings were that the Social Security program not only increases the income statuses of both posttransfer and pretransfer poor elderly people considerably, but that the program also significantly equalizes the distribution of income among them. In addition, the SSI program supplements the Social Security program in establishing an even greater safety net, especially for posttransfer poor elderly people. On the other hand, Social Security benefits make the income disparity among races greater between both posttransfer and pretransfer poor elderly people. Implications for policy are discussed. PMID- 12557992 TI - Health care rationing affecting older persons: rejected in principle but implemented in fact. AB - Health care resources are finite and, therefore, need to be rationed among potential users. Over the past decade and a half in the United States, a variety of explicit, official rationing schemes have been proposed, including some in which chronological age would play a significant role. For ethical and political reasons, it is very unlikely that any age-based rationing schemes will be adopted explicitly and officially. However, various de facto forms of health care rationing are occurring at present. This article outlines the implications of payer behavior, physician practice patterns, the development of evidence-based clinical practice parameters or guidelines, and reliance on consumer choice of health plans as unofficial and generally unacknowledged mechanisms of health care rationing that may exert an important impact on the accessibility of health services for older persons. PMID- 12557993 TI - Expansion of long-term care in the prison system: an aging inmate population poses policy and programmatic questions. AB - Throughout the United States, departments of corrections are experiencing increases in their inmate populations. More specifically, the number of aging inmates is increasing and will continue to grow as younger prisoners who have long sentences with no possibility of parole age in prison. In addition, the number of younger inmates with illnesses such as AIDS has increased. Although long-term care can be required by individuals of any age, the need for such assistance tends to increase with age. Long-term care, therefore, can be seen as an issue confronting prisons with aging inmate populations. Yet, little is known about the nature or extent of the need. This paper focuses on older inmates and includes reasons for the increased need for long-term care in the prison setting. The standard for prison health care, the long-term-care status of older inmates, and examples of long-term-care services and facilities are described. Key questions related to furnishing long-term care to an older incarcerated population are identified. Recommendations are presented for both corrections and long-term-care providers and policymakers as they develop strategies to address this challenge. PMID- 12557994 TI - Gerontological perspectives on crime and nuisance: the elderly critically evaluate housing designs in the British city. AB - This paper investigates the perceptions of the elderly in relation to crime and nuisance and the fear of crime associated with stereotypical British housing designs. Demographically, this diverse though highly urbanized group continues to grow; group members' observations, therefore, have increasing social relevance and political importance and are crucial for assessing and informing both current policy and the evolution of future policy initiatives. Crime Prevention Through Environmental Design (CPTED) has become popular once again in America, Australia, Canada, South Africa, as well as in Europe and Britain. A crucial dimension to this theory concerns the perception of "territoriality," "surveillance," and "image" within the design of the built environment derived from Newman's "Defensible Space" concepts (1973). This paper presents and discusses the ways in which the elderly associate crime and nuisance with a range of traditional housing designs. The findings strongly reinforce Newman's theory. The paper concludes that the design and, perhaps more importantly, the management of residential housing influence the perceived levels of crime, nuisance, and fear of crime, and the "defensible" qualities of each specific design. Such perceptions will arguably affect elderly people's ability to maintain their privacy, dignity, and autonomy, their physical and psychological well-being, and their social inclusion. Policy implications for housing the elderly safely within the community are reviewed. PMID- 12557995 TI - Evaluating consumer satisfaction in residential continuing care settings. AB - The outcome of the project reported on here is a client-centered consumer satisfaction questionnaire designed to evaluate new models of residential continuing care in Alberta, Canada. Satisfaction is defined as a multi dimensional construct that is grounded in the consumer's experience. Consultation with the clients of the services during development of the instrument ensured that characteristics important to the clients were assessed. The result is an instrument with which to measure satisfaction that is fully client-centered and that, with appropriate modifications, can be used to monitor any client-centered program for cognitively-able continuing care clients. PMID- 12557996 TI - The telescoping suture--Part 1: Does this technique improve the mechanical behavior of a biomaterial?: Calf pericardium. AB - The authors study the mechanical behavior of calf pericardium employed in the construction of cardiac valve leaflets when subjected to telescoping suture, followed by tensile stress until rupture. One hundred twenty pericardial tissue samples were employed, 60 cut from root-to-apex and another 60 cut in transverse direction. Each of these two groups consisted of 12 control samples that were left unsutured and four sets of 12 samples each that were rejoined by telescoping suture using silk, Prolene, nylon or Gore-Tex., and subjected to tensile stress. At the rupture of the sutured tissues, the tensile stress of the suture materials ranged between 57.54 MPa for the series sewn lengthwise with Gore-tex and 114.08 MPa for the series sewn crosswise with silk. At these levels of stress, the deformation of the suture thread was much less marked than that of the calf pericardium, and internal stresses were produced that were difficult for the biomaterial to absorb. There was a loss of real load in all the sutured series when the observed resistance to rupture, expressed in kilograms, was compared with the estimated value. This loss of resistance did not invalidate the telescoping suture technique since the resistance to rupture was still much greater than that associated with suturing the two edges of the cut pericardium together. This report confirms the deleterious role of the shear force generated in the pericardium by the suture. PMID- 12557997 TI - The telescoping suture--Part II: A novel method to improve the mechanical behavior of a new biomaterial: ostrich pericardium. AB - Ostrich pericardium, sutured using a telescoping or overlapping technique, was studied to determine its mechanical behavior. From each of 12 pericardial sacs, four contiguous strips were cut longitudinally, from root to apex, and another four contiguous strips were cut in transverse direction. One of the strips in each set of four was used as an unsutured control and the remaining three were sutured by overlapping 0.5 cm of the tissue and sewing with Gore-tex, Prolene or Pronova. These 96 samples were then subjected to tensile testing along their major axes until rupture. The tensile stresses recorded in the suture materials at the moment tears appeared in the pericardium ranged between 55.99 MPa and 70.23 MPa for Gore-tex in samples cut in the two directions. Shear stress became ostensible at 56 MPa, with clearly evident tears. However, microfracture of the collagen fibers must be produced at much lower stress levels. The comparison of the resistance in kilograms (machine-imposed), without taking into account the sections in which the load was applied, demonstrated only a slight loss of load when the telescoping suture was employed in ostrich pericardium samples. Ostrich pericardium may continue to be an alternative biological material for the construction of heart valve leaflets. PMID- 12557998 TI - Stimuli sensitive polymethacrylic acid microparticles (PMAA)--oral insulin delivery. AB - This study investigated polymethacrylic acid (PMAA) microparticles for controlled release of Insulin in oral administration. The microparticles were characterised by scanning electron microscopy (SEM) for morphological studies. The swelling behaviour and drug release profile in various pH media were studied. The % swelling of gels was found to be inversely related to the amount of crosslinker added. Inclusion complex of betaCD and Insulin was studied using polyacrylamide gel electrophoresis (PAGE). Optimum complexation was obtained in the ratio 100 mg betaCD: 200 IU Insulin. The release pattern of Insulin from Insulin-betaCD complex encapsulated PMAA microparticles showed release of Insulin for more than seven hours. PMID- 12557999 TI - Evaluation of electrostatically endothelial cell seeded expanded polytetrafluoroethylene grafts in a canine femoral artery model. AB - PURPOSE: The purpose of this study was to evaluate the extent (luminal coverage) of the endothelial cell (EC) lining/neointimal development and the thromboresistance of electrostatically EC seeded small diameter e-PTFE vascular grafts. METHODS: This evaluation consisted of harvesting autologous, canine jugular vein ECs, electrostatically EC seeding the e-PTFE grafts (4 mm GORE-TEX, Length = 6 cm), implanting the grafts in a canine femoral artery model for six weeks, and excising the graft for histological and scanning electron microscopy evaluations. RESULTS: The results of the histological evaluation (mid-graft region only) indicated that the electrostatic EC seeding significantly affected neointimal development (p < 0.01) and the degree of thrombus formation (p < 0.001) within the EC seeded grafts versus the untreated control grafts. Scanning electron microscopy examination demonstrated a mature, confluent endothelium with a "cobblestone" appearance on the EC seeded graft luminal surface. The control grafts demonstrated an equal distribution of SMCs through the graft wall while the electrostatically EC seeded graft sections exhibited an uneven SMC cellular distribution which was skewed toward the graft luminal surface. CONCLUSIONS: The presence of electrostatic EC seeding significantly (p < 0.01) enhanced the development of a neointima and reduced the incidence of thrombosis in e-PTFE grafts implanted in a canine femoral artery model. Results of the mid-graft SMC migration measurements indicate that the electrostatic EC seeding had a significant (p < 0.001) impact on the acute healing of the standard wall e-PTFE vascular graft specimens. PMID- 12558000 TI - Anticoagulant activity of immobilized heparin on the polypropylene nonwoven fabric surface depending upon the pH of processing environment. AB - Antenna coupling microwave plasma enables a highly oxidative treatment of the outmost surface of polypropylene (PP) nonwoven fabric within a short time period. Subsequently, grafting copolymerization with acrylic acid (AAc) makes the plasma treated fabric durably hydrophilic and excellent in water absorbency. With high grafting density and strong water affinity, the pAAc-grafted support greatly becomes feasible as an intensive absorbent and as a support to promote heparin immobilization through amide bonds. For heparin immobilized in acidic condition, the carbonate groups of the molecule tend to dissolve and passive encapsulation of the molecule prevents its functional groups from bonding with the carboxylic acid of pAAc. This effect leads to inhibit the immobilization process and consequently reduces the quantity as well as the bioactivity of the immobilized heparin. In alkaline processing environment, the oxidized uronic acid residues in heparin-related glycans are presumably cleaved and the removal of some oxidized residuals before immobilization process is likely to reduce the chain length of heparin. In the latter case, anticoagulant Factors X and XII, but not thrombin, are unaffected. Anticoagulant activity test using activated partial thromboplastin time (aPTT) is more sensitive in assessing heparin-immobilized surfaces, since it corresponds to Factor X and initiates the inhibition of Factor XII and thrombin. Likewise, platelets adhesion on the surfaces decreases as the process shifted from acidic to alkaline condition, whereas the hydrophilic character of the grafted pAAc markedly contributes to extend physical insertion of platelets. The immobilized heparin has a great part of original bioactivity, depending on the pH of the processing environment and the immobilized quantity. Relative bioactivity based upon aPTT tests is partially held longer than 90 days for the sample prepared in the alkaline or neutral environment. PMID- 12558001 TI - Overcoming barriers and creating opportunities to reduce burden of affective disorders: a new research agenda. PMID- 12558002 TI - Burden of illness. AB - The burden of affective disorders includes costs and the pain and suffering of affected individuls. Burden can be perceived from social and private perspectives. Although no ideal measure of burden exists, ample evidence documents the extensive cost and other negative impacts of affective disorders. Affective disorders are associated with disruptive family relations, higher health care costs for comorbid conditions, elevated rates of suicide, and lower productivity. Reserch should focus on improving measures of burden in general and on quantifying burden from the standpoint of diverse population groups. PMID- 12558003 TI - Barriers to reducing burden of affective disorders. AB - This paper summarizes 3 sets of barriers to reducing burden of affective disorders including factors that contribute to 1) the risk, course, and outcomes of affective disorders; 2) help-seeking and use of health and mental health services for affective and other mental disorders; and 3) the appropriateness of treatments used for affective disorders. On the basis of this review, the authors recommend research needed to identify modifiable barriers to reducing the burden of affective disorders and to identifying opportunities to reduce these barriers. This new research should focus on clarifying societal, family, and consumer, clinician, and system barriers to recognizing disorders, seeking and providing care, and adhering with guideline concordant care, and should include barriers that apply to both treatment and prevention services. PMID- 12558004 TI - Practice-based interventions. AB - Current evidence indicates there remains a large gap in the provision of depression care, particularly in primary care. Several studies have demonstrated that interventions based on the chronic disease management model can improve patient outcomes. Challenges include designing more robust interventions that can move easily into a wide variety of primary care organizations. More research is needed to develop programs to improve outcomes for children with depression and adults with bipolar disorders. PMID- 12558005 TI - Community-based interventions. AB - This paper explores the potential of community-based, public health-oriented interventions as a tool for reducing the burden of affective disorders on individuals, their families, and communities. The paper reviews the use of community-based interventions with other health-related problems and describes potential applicability for affective disorders such as changing public attitudes, reducing social stigma, facilitating access, or supporting treatment adherence for populations in their community settings. An agenda for developing this field of intervention research is proposed. PMID- 12558006 TI - Policy intervention. AB - This paper addresses market failure due to externalities, as well as information asymmetries and public policy problems that need to be solved to ensure high quality care for affective disorders. We delineate the problems in parity legislation, managed care, as well as Medicare and Medicaid that need to be addressed to reduce the burden of illness affective disorders. A research agenda is developed for formulating and implementing public policy. PMID- 12558007 TI - Research on reducing burden of affective disorders for special populations: introduction and general recommendation. AB - Populations should be prioritzed for research on reducing the burden of affective disorders based on expected or known burden or current level of unmet need for care. We recommend a focus on ethnic minorities, impoverished women, children, adolescents, and individuals with bipolar illness. Studying effectiveness of interventions and the efficiency of delivery systems are priorities for special popultions. PMID- 12558009 TI - Ethnic minorities. AB - Ethnic minorities have relatively similar rates of mood disorders as do white Americans, but they are much less likely to receive appropriate care. Barriers to care include lack of insurance, few minority providers' racism, and distrust of care providers. A priority in research is identifying practice interventions and policies that could eliminate disparities in care. PMID- 12558008 TI - Bipolar disorder. AB - Manic-depressive (bipolar) disorder is a severe, relapsing mental illness that shares characterstics both with major depressive disorder and with serious mental illnesses such as schizophrenia. Like schizophrenia, it is a chronic disorder, and is treated primarily in the specialty mental health sector. Rates of appropriate treatment are low. Functional outcome is compromised for the majority of individuals who have this disorder. Societal costs are exceeded only by those for schizophrenia. Existing cost calculations likely underestimate societal costs because of underestimating functional impact and neglecting to account for the substantial proportion of individuals who are institutionalized outside of the health care system (e.g., in prison). Little is known as yet regarding manic depressive disorder in historically underserved groups and in vulnerable groups such as the elderly. There are major lacunae with regard to this disorder in the grant portfolios of all federal agencies mandated to address the needs of Americans with serious mental illnesses. The authors in the context of the Wider NIMH Affective Disorders Workgroup propose several specific recommendations to address the needs of this costly and underresearched disorder. PMID- 12558010 TI - Children and adolescents. AB - Until the 2001 Surgeon General's report there had been no acknowledgment of the need for a national plan for research priorities to improve services and reduce illness burden for children and adolescents with severe mental disorders. Barriers to services among those in need include individual and family factors, and clinician and service system factors. Additionally, little reserch is available on the impact of major policy reforms on children's ability to obtain efficacious care. Critical research gaps exist in a number of areas with the prevention and early intervention area representing a particularly important missed opportunity. PMID- 12558011 TI - The elderly. AB - Late life depression is common and associated with disability, reduced quality of life, mortality, and high health care costs. Depressed older adults frequently have comorbid medical illnesses and cognitive impairment, but relatively little is known about the diagnosis and treatment of depression in the face of these comorbid conditions. Only a minority of depressed older adults receive specialty mental health care and most depression care is provided in primary care where few receive effective treatment. Very little is known about the epidemiology and quality of care for bipolar disorder in late life. Additional research should focus on the quality and outcomes of care for older adults with affective disorders in diverse settings (including primary care, specialty mental health care, home health care, nursing homes, and assisted living facilities) and on the care of older adults who have affective disorders and comorbid medical disorders, dementia, substance use disorders, or chronic pain. PMID- 12558012 TI - Gender issues and socially disadvantaged women. AB - Gender differences in overcoming barriers to mental health care for affective disorders have not been studied. Further, the role of mental health care for parents as it affects child outcomes is important but currently not well studied. In particular, care for disadvantaged young women is an area in need of further research. PMID- 12558013 TI - Research development mechanisms. AB - Studies designed to reduce the burden of affective disorders should apply and develop theories and methods from diverse social sciences that could strengthen current interventions. A series of papers from diverse fields, such as quality engineering, behavioral economics, etc. might be a needed first step. Methodological research on design strategies such as group-level randomized trials, or instrumental variables analyses are needed. Finaly, qualitative studies to understand diverse stakeholder views are also needed. To pursue these areas, interdisciplinary training programs are needed to develop skilled researchers to study communities and community-based delivery settings. New research infrastructures are needed to support community and research collaborations, as well as supporting development of new technologies to enable diffusion of care. PMID- 12558014 TI - Parental problem recognition and child mental health service use. AB - This study estimates the prevalence and correlates of two components of problem recognition among parents and assesses their relative effects on child mental health service use in several settings. Analyses were based on data from a population-based sample of 1,420 youth-parent pairs. Child psychopathology and impairment were assessed using the Child and Adolescent Psychiatric Assessment. Problem perception was defined as reporting one or more problems or needs; family impact as reporting one or more impacts. Recent use of 30+ types of mental health services was examined. The frequency of problem perception was 13.3% and family impacts 11.2% across all observations. Among parents of children with 1+ DSM-IV psychiatric diagnosis, 39.0% perceived problems and 31.7% perceived impacts. The strongest predictor of problem perception was impact and vice versa. Problem perception (and not impact) was predictive of specialty services after controlling for child illness. Neither problem recognition component predicted general medical or school use. Findings suggest the need for parent education to help them identify serious problems and for universal screening to ensure that access to specialty services is not dependent solely on parents. Problem recognition should be expanded to include perceptions of other adults in models of access mental health care. PMID- 12558015 TI - Moderators and mediators of client satisfaction in case management programs for clients with severe mental illness. AB - This randomized experiment first determined that clients with severe mental illness who received Assertive Community Treatment (ACT) were more satisfied with their treatment program than were clients in a brokered case management program (BCM). Eight demographic and diagnostic variables were examined as potential moderators of the treatment effect. Only one of the eight varables, diagnosis of depression, interacted with treatment condition to effect client satisfaction. However, there were several important mediators of client satisfaction: intensity of program contacts, continuity of program contacts, supportive services (i.e, assistance with activities of daily living), and the strength of the helping alliance as perceived by the client. PMID- 12558016 TI - Biomedical and biological mass spectrometry. AB - This review focuses on biological and biomedical mass spectrometry, and covers a selection of publications in this area included in the MEDLINE database for the period 1987-2001. Over the last 15 years, biological and biomedical mass spectrometry has progressed out of all recognition. The development of soft ionization methods, such as electrospray ionization and matrix-assisted laser desorption ionization, has mainly contributed to the remarkable progress, because they can easily produce gas-phase ions of large, polar, and thermally labile biomolecules, such as proteins, peptides, nucleic acids and others. The innovations of ionization methods have led to remarkable progress in mass spectrometric technology and in biochemistry, biotechnology and molecular biology research. In addition, mass spectrometry is one of the powerful and effective technologies for drug discovery and development. It is applicable to studies on structural determination, drug metabolism, including pharmacokinetics and toxicokinetics, and de novo drug discovery by applying post-genomic approarches. In the present review, the innovative soft ionization methods are first discussed along with their features. Also, the characteristics of the mass spectrometers which are active in the biological and biomedical research fields are also described. In addition, examples of the applications of biological and biomedical mass spectrometry are provided. PMID- 12558017 TI - Integration of chemical and biochemical analysis systems into a glass microchip. AB - This review focuses on the integration of chemical and biochemical analysis systems into glass microchips for general use. By combining multiphase laminar flow driven by pressure and micro unit operations, such as mixing, reaction, extraction and separation, continuous-flow chemical processing systems can be realized in the microchip format, while the application of electrophoresis-based chip technology is limited. The performances of several analysis systems were greatly improved by microchip integration because of some characteristics of microspace, i.e., a large specific interface area, a short molecular diffusion time, a small heat capacity and so on. By applying these concepts, several different analysis systems, i.e., wet analysis of cobalt ion, multi-ion sensor, immunoassay, and cellular analysis, were successfully integrated on a microchip. These microchip technologies are promising for meeting the future demands of high throughput chemical processing. PMID- 12558018 TI - M-DNA: a self-assembling molecular wire for nanoelectronics and biosensing. AB - M-DNA is a complex between divalent metal ions such as Zn2+ and duplex DNA which forms at pH 8.5. Unlike B-DNA, M-DNA does not bind ethidium so that M-DNA formation can be monitored conveniently by an ethidium fluorescence assay. M-DNA was shown to be a better conductor than B-DNA by fluorometric measurements of electron transport in donor-acceptor labelled duplexes; by direct conductivity measurements of M-DNA bound between gold electrodes and by cyclic voltammetric studies on ferrocene labelled duplexes attached to gold microelectrodes. As is the case with B-DNA, M-DNA can self-assemble into a variety of structures and is anticipated to find widespread use in nanoelectronics and biosensing. PMID- 12558019 TI - Effects of viability and lectin protein binding on dielectrophoretic behavior of single yeast cells. AB - The dielectrophoretic (DEP) behavior of individual yeast cells (5-7 microm in diameter) in aqueous media was observed in a fabricated planar quadrupole microelectrode with a working area of 100 microm in diameter by an optical microscope. The yeast cells migrated in the radial direction in the working area. The DEP velocity of the cells increased as they approached the electrode. The DEP trajectory of the cells was analyzed with a theoretical equation derived previously, and the dielectrophoretic mobility was determined. The dielectrophoretic mobility was found to be affected by the viability of cells, the conductivity of the medium, and the binding of lectin protein (concanavalin A) to the cell surface. These DEP behaviors were analyzed based on the permittivities and conductivities of the cell interior and wall, and those of the medium. PMID- 12558020 TI - Simultaneous measurement of the migration velocity and adsorption force of micro particles using an electromagnetophoretic force under a high magnetic field. AB - A simultaneous measurement technique for determining the migration velocity of a micrometer-sized particle in a capillary and the adsorption force to the inner surface of the capillary has been proposed. This technique is based on an electromagnetophoretic force being exerted on a micro-particle in an electrolyte solution, which is governed mainly by the electromagnetic buoyancy, when a homogeneous magnetic field is applied at a right angle to the electric current through the medium. By the electromagnetic buoyancy, micro-particles such as polystyrene, carbon and yeast were migrated perpendicular to the direction of the electric current and reached a fused-silica wall. A switching of the current direction could desorb the particle from the wall, and allowed to calculate the detaching force from the desorbing current. The migration velocity normalized to the size in the magnetic field of 10 T was increased in the order of yeast, carbon and polystyrene, while reflecting the decreasing order of the apparent conductivity of the particles. The desorption force could be measured up to 1 nN with a sensitivity of pN. The observed interaction forces of polystyrene and carbon were in the range of 250-600 pN with large deviations. PMID- 12558021 TI - Highly stereoselective, uniformly sized molecularly imprinted polymers for cinchona alkaloids in hydro-organic mobile phases. AB - Highly stereoselective, uniformly sized molecularly imprinted polymers (MIPs) for cinchona alkaloids, cinchonine (CN) and cinchonidine (CD), were prepared using methacrylic acid (MAA) as a functional monomer and ethylene glycol dimethacrylate (EDMA) as a cross-linker. The MIPs were evaluated using a mixture of phosphate buffer and acetonitrile as the mobile phase. The CN- and CD-imprinted MAA-co-EDMA polymers can recognize the respective template molecule more than the other diastereomer, and afford an excellent diastereomer separation of CN and CD. In addition, the MIPs gave diastereomer separations of structurally related compounds, quinidine and quinine. The retentive and stereoselective properties of those compounds on the MIPs suggest that electrostatic and hydrophobic interactions can work to recognize these compounds. Furthermore, thermodynamic studies reveal that the entropy-driven effect is significant at mobile-phase pH 5.4, while the enthalpy-driven interactions seem to be dominant at mobile-phase pH 9.6. PMID- 12558022 TI - Nano-kinetics of probe-particles in solution visualized by a pin-fiber video scope. AB - The nano-kinetics of colloidal particles and living cells with the colloidal particles were visualized by a newly developed video scope. The system of the new video scope has a feature of fine controlling the illumination conditions by using a single optical fiber. This characteristic enables one to obtain clear images of living cells and the motions of colloidal particles by light-scattering effects. In the experiments, RBL-2H3 cells and gold colloidal particles were observed. Scattering images with high contrast and a dark background like in dark field observations could be attained. In the experiments, a pulsed laser was also applied. The results obtained in this study could validate the effectiveness and possibility of a new video scope for applications to biological and biomedical fields. PMID- 12558023 TI - Development of the single-cell MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass-spectroscopic assay. AB - MALDI-TOF mass spectrometry was used to detect intracellular molecules from a single intact cell on monolayers of other cells. Intracellular molecules, e.g., histamine, were gradually increased in a mouse bone marrow-derived mast cell by a maturation process. A single cell was captured by a microsuction pipette, and the mass spectra of intracellular histamine were measured directly. Finally, the time course of the intracellular molecular contents and the maturation stage from a single cell were estimated. PMID- 12558024 TI - A glass capillary microelectrode based on capillarity and its application to the detection of L-glutamate release from mouse brain slices. AB - A new glass capillary microelectrode for L-glutamate is described using pulled glass capillaries (tip size, approximately 12.5 microm) with a very small volume (approximately 2 microl) of inner solution containing glutamate oxidase (GluOx) and ascorbate oxidase. The operation of the electrode is based on capillary action that samples L-glutamate into the inner solution. The enzyme reaction by GluOx generates hydrogen peroxide that is detected at an Os-gel-HRP polymer modified Pt electrode in a three-electrode configuration. The amperometric response behavior of the electrode was characterized in terms of the capillarity, response time, sensitivity and selectivity for measurements of L-glutamate. The currents at 0 V vs. Ag/AgCl increased linearly with the L-glutamate concentration from 10 to 150 microM for in vitro and in situ calibrations. The response was highly selective to L-glutamate over ascorbate, dopamine, serotonin and other amino acids. The detection of L-glutamate in the extracellular fluids of different regions of mouse hippocampal slices under stimulation of KCl was demonstrated. PMID- 12558025 TI - Simultaneous determination of glucose and L-lactate in rat brain by an electrochemical in vivo flow-injection system with an on-line microdialysis sampling. AB - An electrochemical in vivo flow-injection system with an on-line microdialysis sampling is proposed for the simultaneous monitoring of L-lactate and glucose in rat brain. In the first stage of the operation, the dialysate from the microdialysis probe is delivered to a sample loop of the six-way autoinjector by perfusing Ringer's solution for 80 s at 5 microl min(-1). In the second stage, the dialysate collected in the sample loop is automatically injected for 10 s into the flow-injection line. Injected dialysate is split into two streams and two portions pass through two channels with two different immobilized enzyme reactors (glucose oxidase and lactate oxidase immobilized reactors) to produce hydrogen peroxide from glucose and L-lactate in the dialysate. After a subsequent confluence of the streams, produced hydrogen peroxide can be detected amperometrically at a downstream poly(1,2-diaminobenzene) film-coated platinum electrode, without any interference from oxidizable species and proteins present in the dialysate. Because each channel has a different residence time, two peaks are obtained. The first peak corresponds to L-lactate and the second peak to glucose. The peak current is linearly related to the concentrations of L-lactate between 0.2 and 10 mM and glucose between 0.1 and 20 mM. The present method can be successfully applied to the simultaneous in vivo monitoring of L-lactate and glucose in rat brain. The analytical speed is 45 dialysates h(-1). PMID- 12558027 TI - Oligodeoxynucleotide-modified capillary for electrophoretic separation of single stranded DNAs with a single-base difference. AB - We describe here a method of affinity capillary electrophoresis in which oligodeoxynucleotide (ODN) was immobilized onto the inner surface of the capillary. The immobilized ODN functioned successfully as an affinity ligand for sequence-based DNA separation. Six- or 12-mer ODN with a sequence complementary to one of the c-K-ras gene was used as an immobilized ligand. When the 12-mer ODN was used, the detection peak for the complementary ODN disappeared selectively, while the single-base mutant was detected as usual. In contrast, when the 6-mer ODN was used as the affinity ligand with a mixture of the complementary ODN and its single-base mutant, it was possible to detect both as completely separate peaks. That is, the separation mode was dependent on the base number of the immobilized ODN used as an affinity ligand. PMID- 12558026 TI - In vivo imaging of brain dopaminergic neurotransmission system in small animals with high-resolution single photon emission computed tomography. AB - High-resolution single photon emission computed tomography (SPECT) provides a unique capability to image the biodistribution of radiolabeled molecules in small laboratory animals. Thus, we applied the high-resolution SPECT to in vivo imaging of the brain dopaminergic neurotransmission system in common marmosets using two radiolabeled ligands, [123I]2beta-carbomethoxy-3beta-(4-iodophenyl)tropane (beta CIT) as a dopamine transporter (DAT) ligand and [123I]iodobenzamide (IBZM) as a dopamine D2 receptor (D2R) ligand. Specific images of the striatum, a region with a high density of dopaminergic synapses, were obtained at 240 min and 60 min after injection of [123I]beta-CIT and [123I]IBZM, respectively. Furthermore, a significantly low accumulation of [123I]beta-CIT in the striatum was observed in MPTP-treated animals compared with results for a control group, and a similar accumulation in the control group was observed with the pretreatment of deprenyl in the MPTP-treated animals. However, the striatal accumulation of [123I]IBZM showed no changes among the control, MPTP-treated, and deprenyl-MPTP-treated groups. These SPECT imaging results agreed well with those of DA concentration and motor behavior. Since MPTP destroys nigrostriatal dopamine nerves and produces irreversible neurodegeneration associated with Parkinsonian syndrome, SPECT imaging data in this study demonstrated that deprenyl shows its neuroprotective effect on Parkinsonism by protecting against the destruction of presynaptic dopamine neurons. PMID- 12558028 TI - Direct detection of single nucleotide polymorphism (SNP) with genomic DNA by the ferrocenylnaphthalene diimide-based electrochemical hybridization assay (FND EHA). AB - A ferrocenylnaphthalene diimide-based electrochemical hybridization assay (FND EHA) was applied to the direct detection of a C-to-G transition in a codon (TCA) for Ser-447 of the human lipoprotein lipase (LPL) gene, which resulted in the termination of the LPL protein there. Either one of two 13-meric oligonucleotide probes, S447 WT and S447X MT, representing sequences complementary to those of the wild type (WT) and mutated (MT) forms, was immobilized on a gold electrode, followed by hybridization with chromosomal DNA extracted from human leukocytes under the condition in which both WT- and MT-type sequences can form a duplex. These two electrodes were soaked in an electrolyte containing FND under a condition [0.1 M HOAc/KOAc (pH 5.6) containing 0.1 KCl and 0.05 mM FND at 40 degrees C], in which only the MT duplex could undergo dissociation. FND was concentrated in proportion to the amount of the duplex formed on the electrode to give rise to a current signal. The electrochemical signal ratios obtained for WT/WT, WT/MT and MT/MT were close to the theoretical 2:1:0 with the S447 WT modified electrode, and was again close to 0:1:2 with the S447X MT-modified one. PMID- 12558029 TI - Targeted proteo-glycomics analysis of Sialyl Lewis X antigen expressing glycoproteins secreted by human hepatoma cell line. AB - Sialyl Lewis X (SLEX) antigen, Neu5Acalpha2-3Galbeta1-4 (Fucalpha1-3) GlcNAc-R, plays important roles in cell-to-cell interaction: for example, the E- and P selectin-mediated influx of SLEX expressing leukocytes into inflamed areas. A human hepatocellular carcinoma cell line, HepG2 cells, was highly expressed SLEX on secreted glycoproteins and cell surface, in contrast with HuH-7 cells. We identified SLEX expressing glycoproteins in HepG2 cultured medium by two dimensional polyacrylamide gel electrophoresis, followed by in gel digestion and peptide mass fingerprint using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOFMS), including transferrin, alpha1 antitrypsin, alpha2-HS glycoprotein and beta-glycoprotein. We analyzed N-glycans of these glycoproteins by MALDI-TOFMS in combination with exoglycosidase digestion; our results indicate increases in poly-fucosylated and high-branched N glycans. High alpha1,3-fucosylation in glycoproteins would be caused by increased expression of alpha1,3-fucosyltransferase activities in HepG2 cells. PMID- 12558030 TI - Convenient analytical methods for endo-type glycosidase that acts on glycoconjugates and their application in glycotechnology. AB - The following procedures were established in order to develop useful degradation enzymes of glycoconjugate for developing postgenome and postproteome research: (1) Enzyme activity with a short time reliability was measured using small amounts by HPLC. (2) The structures of the sugar chains liberated from the glycoconjugate were non-destructively analyzed using small amounts of sugar chains only by 1D 1H-NMR and H-H COSY spectrometry and a computer simulation of the spectrum. (3) The conformations of the sugar chains liberated from a glycoconjugate in aqueous solution were estimated using 1D 1H-NMR and H-H COSY spectrometry and the anisotropic effect. Endo-alpha-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242 developed using the above methods transferred the sugar chain to sugars and peptides; therefore, it was also an effective enzyme when synthesizing sugar chains and glycopeptides. PMID- 12558031 TI - Sensitive and high-throughput analyses of purine metabolites by dynamic pH junction multiplexed capillary electrophoresis: a new tool for metabolomic studies. AB - On-line sample preconcentration by a dynamic pH junction in conjunction with multiplexed capillary electrophoresis (CE) and UV detection represents a sensitive and high-throughput format for future metabolomic research, such as purine analysis. The optimization of purine focusing can be rapidly assessed by systematically altering the sample matrix properties, such as the buffer co-ion, pH and ionic strength using a 96-capillary array format. This method permits focusing of large sample injection volumes, resulting in over a 50-fold improvement in the concentration sensitivity. The limit of detection (S/N = 3) for purine metabolites was less than 8.0 x 10(-8) M under optimum conditions when using UV absorbance. Dynamic pH junction multiplexed CE demonstrated excellent linearity over a hundred-fold concentration range, as well as low inter-capillary precision in terms of normalized migration times and peak areas. The potential for clinically relevant high-throughput analyses of micromolar amounts of purine metabolites in urine was also demonstrated. PMID- 12558032 TI - Highly sensitive simultaneous bioluminescent measurement of acetate kinase and pyruvate phosphate dikinase activities using a firefly luciferase-luciferin reaction and its application to a tandem bioluminescent enzyme immunoassay. AB - We have developed a simultaneous bioluminescent measurement of acetate kinase (AK) and pyruvate phosphate dikinase (PPDK) activities and its application to a tandem enzyme immunoassay. The principle of the proposed assay is as follows. In the first step, AK generates ATP from ADP and acetylphosphate, and the ATP is determined by the firefly luciferase-luciferin reaction. In the second step, the bioluminescent intensity from AK is eliminated by adding glucose and ADP dependent hexokinase, which forms AMP from ADP. At the same time, the PPDK catalyzes the interconversion of AMP, diphosphate, and phosphoenolpyruvate to ATP, phosphate and pyruvate. The ATP formed by PPDK is also determined by the firefly luciferase-luciferin reaction. The detection limits (at blank + 3SD) of AK and PPDK were 1.03 x 10(-20) and 2.05 x 10(-20) mol per assay, respectively. The method was applicable to a bioluminescent enzyme immunoassay for the assay of insulin and C-peptide in the same sample. PMID- 12558033 TI - The immunoassay of methotrexate by capillary electrophoresis with laser-induced fluorescence detection. AB - Immunoassay is widely employed as a highly sensitive, specific analytical method for hormones and drugs in biological samples. A technique utilizing capillary electrophoresis with laser-induced fluorescence detection was examined based on the reaction process of these immunoassays in order to develop a protocol characterized by high sensitivity and high speed. The conditions of the antigen antibody reaction and capillary electrophoresis were variously examined using fluorescein-labeled methotrexate and the antibody of methotrexate. As a result, the immunoassay could be completed within a few minutes. Moreover, detection in the pg range could be accomplished. The sensitivity corresponded to that of radioimmunoassay. A simultaneous multi-component analysis of the immunoassay is also possible due to the high resolving power of capillary electrophoresis. In this study, the possibility of a simultaneous analysis of methotrexate and vancomycin was also investigated. PMID- 12558035 TI - Lasting chemiluminescence of 3-indoleglyoxylyl chloride and its enhancement. AB - The chemiluminescence (CL) intensities of various indole derivatives substituted with a glyoxylyl group at the 3-position and a hydroxyl group at the 5-position of the indole ring were compared upon the addition of H2O2 in alkaline media. The CL intensities of 3-indoleglyoxylyl chloride, 3-indoleglyoxylic acid, 5 hydroxyindole and 5-benzyloxyindole in CH3CN were 5.9-, 48-, 5.9- and 3.3-fold stronger than that of 3-methylindole. A lasting CL of 3-indoleglyoxylyl chloride was found. Under appropriate conditions, the CL emission reached a maximum within 10 min after the addition of H2O2 in the presence of NaOH, and the intensity was retained for 25 min. One of the final products via the CL reaction of 3 indoleglyoxylyl chloride was indole-3-carboxylic acid. 3-Indoleglyoxylyl chloride emitted light by decompositions via both dioxetane and dioxetanedione. An enhancement effect of beta-cyclodextrin and bovine serum albumin on the CL of 3 indoleglyoxylyl chloride was also found. PMID- 12558034 TI - Speciation of small molecules and inorganic ions in salmon egg cell cytoplasm by surfactant-mediated HPLC/ICP-MS. AB - The speciation of diverse elements in salmon egg cell cytoplasm was performed by a surfactant-mediated HPLC/ICP-MS hyphenated system. In the present experiment, an ODS column coated with CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1 propanesulfonate), which is a zwitterionic bile acid derivative, was employed as a surfactant-mediated separation column, and ICP-MS was used as an element selective detector. The present surfactant-mediated HPLC allowed us to separate large and small molecules within 10 min; large molecules, such as proteins, were eluted within 2.5 min, while small molecules were eluted after 2.5 min, but within 10 min. In the present experiment, Fe, Cu, and Zn in egg cell cytoplasm were observed mostly in species with large molecular weights, indicating that these elements are contained as metalloproteins or metalloenzymes in egg cell cytoplasm. On the contrary, it was found that P, S, Mo, and halogens in egg cell cytoplasm were contained as small molecules or inorganic ions. The major species of P in egg cell cytoplasm was identified as the phosphate ion (PO4(3-)). Molybdenum, Cl, and Br in egg cell cytoplasm were molybdate (MoO4(2-), chloride (Cl-), and bromide (Br-) ions, respectively. PMID- 12558036 TI - Optical cell with a temperature-control unit for a vacuum-ultraviolet circular dichroism spectrophotometer. AB - We constructed an assembled-type MgF2 cell that can function under a high vacuum (10(-4) Pa), and is capable of measuring the vacuum-ultraviolet circular dichroism (VUVCD) spectrum in a wavelength region down to 140 nm for aqueous solutions. Its path length can be adjusted by various spacers in the range from 1.3 to 50 microm. The temperature-control unit of the cell was also constructed with a Peltier thermoelectric element to keep the temperature of a sample within an accuracy of +/- 1 degrees C in the temperature range from -30 to 70 degrees C. The optical cell and the temperature-control unit were confirmed to have good performance by monitoring the VUVCD spectra of ammonium d-camphor-10-sulfonate and myoglobin aqueous solutions. This cell is available not only for VUVCD spectroscopy, but also for vacuum-ultraviolet absorption measurements. PMID- 12558037 TI - Potentiometric and multi-NMR studies of aluminum(III) complex with L-glutamate in acidic aqueous solutions. AB - Complex formation studies of L-glutamate with aluminum(III) ion were conducted in acidic aqueous solutions (pH 2.0 - 5.5) by means of pH-metric titration and multinuclear (1H, 13C and 27Al) NMR techniques. The following results were obtained: (1) Al could weakly coordinate with Glu to form various mononuclear 1:1 (AlLH2+, AlL+, AlLH(-1)) species and dinuclear 2:1 (Al2L4+) species in acidic aqueous solutions, which somewhat agreed with previous findings. (2) The multi NMR spectra of Al-Glu and Al-Asp strongly suggest that, besides negatively charged carboxylate donors (-COO(-), -COO(-)), the amino group of Glu can participate in the binding of Al in the AlL+ and AlLH(-1) species in the case of its deprotonation, which rather agreed with the case of Al-Asp. (3) These tridentate five-+seven-membered joint chelate (-COO(-), -NH2, -COO(-)) complexes exhibit an enhanced stability, which can help to better understand the biological studies that Al-Glu could cross the erythrocyte membrane and the blood-brain barrier (BBB) and be deposited selectively in various brain regions, particularly in the cortex. It will also help to intrinsically understand the Al's role in the biological transamination system, which is a very important process in all living things. PMID- 12558038 TI - Effects on humans elicited by inhaling the fragrance of essential oils: sensory test, multi-channel thermometric study and forehead surface potential wave measurement on basil and peppermint. AB - The effects on humans inhaling the fragrance of essential oils were examined in terms of a sensory test, a multi-channel skin thermometer study and a portable forehead surface electroencephalographic (IBVA-EEG) measurement. The essential oils examined in this study were those of basil and peppermint, because our previous sensory test had indicated an opposite effect of these essential oils when mental work was undertaken; the inhalation of basil produced a more favorable impression after work than before work, whereas peppermint produced an unfavorable impression under these circumstances. For subjects administered basil or peppermint before and after mental work using an inhalator, a series of multi channel skin thermometer studies and IBVA-EEG measurements were conducted. Using such paired odorants, our results showed that when compared between before and after mental work assigned to subjects: (1) the inhalation of basil, in which a favorable impression was predominant on the whole in terms of the sensory evaluation spectrum, was shown to be associated upward tendency in finger-tip skin temperature; (2) whereas these situations were opposite in the case of peppermint, in which the reversed (unfavorable) feature in sensory profiling was accompanied by a decrease in the magnitude of beta waves and a decrease in the finger-tip skin temperature both based on Welch's method, even at p < 0.01, implying a decreasing propensity of the aroused state and of the arousal response. The elucidation of such sensory and physiological endpoints of paired odorants would be of primary importance for human chemoreception science, because these are only rarely recorded during the same experiments, and this paradigm is highly informative about non-verbal responses to odorants. PMID- 12558039 TI - Major-to-ultratrace elements in bone-marrow fluid as determined by ICP-AES and ICP-MS. AB - The major-to-ultratrace elements in human bone-marrow fluid were determined by ICP-AES (inductively coupled plasma atomic emission spectrometry), and ICP-MS (inductively coupled plasma mass spectrometry). The bone-marrow fluid sample was centrifuged prior to acid digestion to exclude the bone piece from bone marrow, and then digested with nitric acid. As a result, 20 elements could be determined over the concentration range from 1610 microg g(-1) for Na to 0.00043 microg g( 1) for W. It was found that Fe, Zn and Sb were enriched by ca. 264-, 7- and 15 fold, respectively, in bone-marrow fluid, compared to those in human blood serum. Alkali metals (K, Rb, Cs), except for Na, were also significantly enriched in bone-marrow fluid. Furthermore, the concentrations of various elements, such as Fe, P, Al, Zn, Cu, Se, Zr, Sn, Ag and W, were significantly higher than those in open seawater. PMID- 12558040 TI - Determination of trace amounts of bisphenol A in urine by negative-ion chemical ionization-gas chromatography/mass spectrometry. AB - We improved an analytical method for determining trace amounts of bisphenol A (BPA) in urine. BPA was subjected to enzymolysis and then to solid phase extraction with a C18 cartridge. The extract was eluted with methanol, and the eluate was concentrated under a nitrogen stream, and then pentafluorobenzylized in an alkali solution. The obtained pentafluorobenzylized compound was purifed using a florisil cartridge, followed by a determination using NCI-GC/MS. This method exhibited an excellent selectivity and reproducibility with a determination limit of 0.1 ng/ml. PMID- 12558041 TI - Electrochemical flow enzyme immunoassay by means of a needle-shaped sampler/reactor. AB - A needle-shaped sampler/reactor was developed for an electrochemical enzyme immunoassay with the direct sampling of living sample blood. This device was evaluated using IgG determination chemistry. Antibodies were immobilized on an inner wall of the sampler/reactor. Incubation for the enzyme reaction was not needed because this reactor was very small (250 microm in diameter). The analysis was conducted within 15 min in the simplest protocol including the reactor refreshment. The limit of detection was 3 pg, and 20 attomol in the most sensitive protocol. Furthermore, the sampling of a solution contained in an agar block and a whole-blood analysis were demonstrated. PMID- 12558042 TI - Analysis method of the angiotensin-I converting enzyme inhibitory activity based on micellar electrokinetic chromatography. AB - A micellar electrokinetic chromatography (MEKC) method was developed for estimating the angiotensin-I converting enzyme (ACE) inhibitory activity by separating the hippuric acid liberated in the ACE reaction mixture in the presence of an inhibitor, captopril. The hippuric acid was successfully separated and detected by MEKC with a 25 mM sodium dodecyl sulfate solution in a 25 mM phosphate-50 mM borate buffer at pH 7.0; the total analysis took about 5 min. A good linear relationship was observed between the inhibitor and the peak area of hippuric acid release. No significant difference in the ACE inhibitory activity (IC50) of captopril (an antihypertensive medicine) or autolyzed-mushrooms (functional foods) was observed between the conventional method and the MEKC method. The MEKC method was found to be a useful technique for a rapid assay of the ACE inhibitory activity. PMID- 12558043 TI - Analysis of reaction products of morphine and codeine with hydrogen peroxide by high-performance liquid chromatography/mass spectrometry. AB - Changes in the chemical structures of morphine and codeine in the presence of hydrogen peroxide, a major component of hair dye and decolorant treatments, were examined with high-performance liquid chromatography/mass spectrometry (LC/MS). A mixture of morphine and hydrogen peroxide solution, after incubation at 39 degrees C for 24 h, produced two reaction products (hydroxymorphines). When codeine was used in place of morphine, one reaction product (hydroxycodeine) was produced, in which the benzene ring was hydroxylated. PMID- 12558044 TI - Fluorescence labeling of oligosaccharides useful in the determination of molecular interactions. AB - A simple method to label oligosaccharides with a multifunctional fluorescent group was developed. Oligosaccharides were quantitatively labeled at their reducing termini with pyrene butanoic acid hydrazide. The pyrene-labeled oligosaccharides were successfully applied to fluorescence polarization measurements and ELISA at picomole quantity, which was not previously reached by other procedures. This labeling method should prove to be useful in a variety of aspects in glycobiology. PMID- 12558045 TI - Novel observation of a circular dichroism band originating from amyloid fibril. AB - A peptide fragment of a non-amyloid-beta component (NAC(1-13)) was studied by CD and electron microscopy. Typical amyloid fibrils were observed by EM in a solution of NAC(1-13). In addition to the beta-structural CD band in the far-UV region, a novel CD band near 285 nm was observed in a peptide solution of NAC(1 13). Taking the NAC(1-13) to contain neither an aromatic amino acid residue nor cystine into account, the CD band can be attributed to amyloid fibrils of NAC(1 13). PMID- 12558046 TI - Range of separation of potential tool for bioseparation, microchip electrophoresis system, for DNA polymorphisms on the human Y-chromosome. AB - For the requirement of a high, fast and sufficient technology to suit the needs of 21st century biotechnology, the separation range of a microchip electrophoresis system was studied. Two DNA fragments on the human Y-chromosome, SY594 (82 bp) and 12f2 (88 bp), were successfully separated with a reproducibility of 1.9% and an accuracy of 2.8%. Then, a mixture of 10 DNA markers ranging from 61 bp to 189 bp was successfully separated with high resolution. All of these results demonstrate the superiority of microchip electrophoresis as a tool for 21st century bioseparation. PMID- 12558047 TI - Self-assembled DNA-conjugated polymer for novel DNA chip. AB - We developed DNA-conjugated polymer for DNA chip fabrication. A 30 mer probe DNA and disulfide bridges were covalently attached to the polymer side chain. The DNA conjugated polymer can be specifically adsorbed on a gold substrate surface by a self-assembly technique. The interaction between fully matched DNA and DNA conjugated polymer was investigated by surface plasmon resonance (SPR) technique. The DNA-conjugated polymer-modified gold surface highly recognized fully matched DNA, rather than unmatched DNA. Therefore, DNA-conjugated polymer can be used for novel DNA chip fabrication. PMID- 12558048 TI - Design of peptide that recognizes double-stranded DNA. AB - A novel molecular tool for double-stranded (ds) DNA detection using synthetic peptide is described. The peptide was designed based on the DNA binding domain of the lambda phage CRO repressor (CRO). The designed peptides contain helix-turn helix (HTH), which is DNA binding motif. A cyclic peptide and a mutant peptide based on CRO were also designed, and the resulting affinity for dsDNA was increased. Furthermore, native amino acids of the peptide were replaced with arginine to increase the affinity for dsDNA. The affinity of these peptides for DNA binding was assessed by surface plasmon resonance (SPR) technique. PMID- 12558049 TI - Evaluation of the molecular recognition of peptide-conjugated polymer. AB - We have recently reported on dodecamer peptides (HPPMDFHKAMTR, CHPQPLKSRNPL) which recognize 52-58th and 197-203rd amino acid sequences of glucose oxidase (GOx) by screening via a phage random peptide library. In this study, a side chain protected peptide monomer (PPM) was synthesized using two peptides (HPPMDFHKAMTR, SHPQPLKSRNPL) and acryloyl chloride. The peptide-conjugated polymer (PCP) was copolymerized with PPMs and N,N-diethylacrylamide (DEAA). The affinity of PCPs to GOx was estimated using surface plasmon resonance detection. This study suggests that PCP is a valuable molecular recognition biomolecule. PMID- 12558051 TI - Purification of lipoxygenase from Chlorella: production of 9- and 13 hydroperoxide derivatives of linoleic acid. AB - Oxygenation of linoleic acid by the enzyme lipoxygenase (LOX) that is present in the microalga Chlorella pyrenoidosa is known to produce the corresponding 9- and 13-hydroperoxide derivatives of linoleic acid (9- and 13-HPOD, respectively). Previous work with this microalga indicated that partially purified LOX, present in the 30-45 and 45-80% saturated (NH4)2SO4 precipitate fractions, produced both HPOD isomers but in different ratios. It was not clear, however, if the observed activity in the two isolates represented the presence of one or more isozymes. In the present work, LOX isolated from the intracellular fraction of Chlorella by (NH4)2SO4 precipitation (35-80% saturated) was purified by ion exchange and hydrophobic interaction chromatography to apparent homogeneity. Analysis of the purified protein by SDS-PAGE and subsequent native size exclusion chromatography demonstrated that LOX in Chlorella is a single monomeric protein with a molecular mass of approximately 47 kDa. The purified LOX produced both the 9-HPOD and 13 HPOD isomers from linoleic acid in equal amounts, and the isomer ratio was not altered over the pH range of 6 to 9. Optimal activity of LOX was at pH 7.5. PMID- 12558050 TI - Clinical trial results support a preference for using CLA preparations enriched with two isomers rather than four isomers in human studies. AB - CLA mixtures are now commercially available. They differ from each other with respect to their content of CLA isomers and their degree of purification. As a group of natural FA, CLA have been widely assumed to be safe. However, the suspected presence of both impurities and particular isomers might induce undesirable side effects. Despite this potential health risk, only a few CLA preparations have been tested under rigorous conditions for clinical efficacy and safety. Based on the limited results available, it is possible to suggest that preparations enriched in c9,t11 and t10,c12 isomers are preferable for human consumption compared to preparations containing four isomers, in terms both of safety and efficacy. PMID- 12558052 TI - Total synthesis of 2-methoxy-14-methylpentadecanoic acid and the novel 2-methoxy 14-methylhexadecanoic acid identified in the sponge Agelas dispar. AB - The phospholipid FA composition of the Caribbean sponge Agelas dispar was revisited and 40 different FA were identified. Among these a novel 2-methoxylated FA, namely, the anteiso methyl-branched 2-methoxy-14-methylhexadecanoic acid, was identified together with the recently discovered iso methyl-branched 2-methoxy-14 methylpentadecanoic acid and the normal-chain 2-methoxytetradecanoic acid. The structures of the iso and anteiso methyl-branched 2-methoxylated FA were confirmed by total syntheses, which were accomplished in seven steps and in 45 48% overall yields. Other phospholipid FA identified in A. dispar include the unusual methyl-branched 10,13-dimethyltetradecanoic acid, 3,7,11,15 tetramethylhexadecanoic (phytanic) acid, and the 11-methyloctadecanoic acid. In addition, the delta5,9 FA (5Z,9Z)-15-methyl-5,9-hexadecadienoic acid and (5Z,9Z) 5,9-octadecadienoic acid were characterized. These findings establish alternative FA biosynthetic possibilities for these marine organisms. PMID- 12558053 TI - In vitro desaturation and elongation of rumenic acid by rat liver microsomes. AB - Various nutritional studies on CLA, a mixture of isomers of linoleic acid, have reported the occurrence of conjugated long-chain PUFA after feeding experimental animals with rumenic acid, 9c,11t-18:2, the major CLA isomer, probably as a result of successive desaturation and chain elongation. In the present work, in vitro studies were carried out to obtain information on the conversion of rumenic acid. Experiments were first focused on the in vitro delta6-desaturation of rumenic acid, the regulatory step in the biosynthesis of long-chain n-6 PUFA. The conversion of rumenic acid was compared to that of linoleic acid (9c,12c-18:2). Isolated rat liver microsomes were incubated with radiolabeled 9c,12c-18:2 and 9c,11t-18:2 under desaturation conditions. The data indicated that [1 (14)C]9c,11t-18:2 was a poorer substrate for delta6-desaturase than [1 (14)C]9c,12c-18:2. Next, in vitro elongation of 6c,9c,11t-18:3 and 6c,9c,12c-18:3 (gamma-linolenic acid) was investigated in rat liver microsomes. Under elongation conditions, [1-(14)C]6c,9c,11t-18:3 was 1.5-fold better converted into [3 (14)C]8c,11c,13t-20:3 than [1-(14)C]6c,9c,12c-18:3 into [3-(14)C]8c,11c,14c-20:3. Finally, in vitro delta5-desaturation of 8c,11c,13t-20:3 compared to 8c,11c,14c 20:3 was investigated. The conversion level of [1-(14)C]8c,11c,13t-20:3 into [1 (14)C]5c,8c,11c,13t-20:4 was 10 times lower than that of [1-(14)C]8c,11c,14c-20:3 into [1-(14)C]5c,8c,11c,14c-20:4 at low substrate concentrations and 4 times lower at the saturating substrate level, suggesting that conjugated 20:3 is a poor substrate for the delta5-desaturase. PMID- 12558054 TI - Fatty acid mobilized by the vascular endothelial growth factor in human endothelial cells. AB - Release of FFA from membrane phospholipids was observed after incubation of umbilical cord vein-derived endothelial cells (HUVEC) with vascular endothelial growth factor (VEGF). In particular, we found an increase of arachidonate, stearate, and palmitate in a time-dependent manner with a peak at 30 min. The maximum increase was reached by arachidonate (4.4-fold), followed by stearate (2.2-fold) and palmitate (1.3-fold). The arachidonate increase can be ascribed to the activation of phospholipase A2 (PLA2). In fact, cells preincubated with arachidonyl trifluoromethyl ketone, a PLA2 inhibitor, showed a marked reduction in arachidonate mobilization. The role of Ca2+ in PLA2 activation was also investigated. Cells incubated with VEGF in the presence of EGTA showed a marked decrease in arachidonate mobilization, whereas incubation with the calcium ionophore A23187 alone produced an increase in arachidonate, although to a lesser extent compared with the VEGF stimulation. Incubation with A23187 in association with PMA produced the same increase in arachidonate as the VEGF treatment. Mitogen-activated protein kinase (MAPK) activity was also found to increase as a consequence of VEGF stimulation. Taken together, these results suggest that the VEGF-mediated activation of PLA2 in HUVEC is dependent on both MAPK-mediated phosphorylation and Ca2+ increase. Furthermore, the increase in stearate and palmitate likely is brought about by the activation of a pathway involving phospholipase D, phosphatidate phosphohydrolase (PAP), and DAG lipase. In fact, the increase in those FFA was prevented when HUVEC were stimulated with VEGF in the presence of ethanol (which inhibits the formation of phosphatidate), propranolol (a specific inhibitor of PAP), or RHC-80267 (a specific inhibitor of DAG lipase). PMID- 12558055 TI - Olive oil phenolics protect LDL and spare vitamin E in the hamster. AB - An animal feeding trial was conducted to investigate whether olive oil phenolics can act as functional antioxidants in vivo. To this end, hamsters were exposed for a period of 5 wk to a dietary regime with either a phenol-rich extra virgin olive oil or extra virgin olive oil from which phenols were removed by ethanol/water-washing. The original oil used in the high olive phenol diet was also used for the preparation of the low phenol diet in order to keep the FA compositions exactly the same. In addition, the vitamin E content was kept identical in both diets. This careful preparation of the diets was undertaken in order to prevent these factors from influencing the antioxidative status in plasma and LDL. Removal of olive oil phenols was shown to reduce both the vitamin E level in plasma and the resistance of LDL to ex vivo oxidation. The results of this study support the idea that extra virgin olive oil phenols improve the antioxidant defense system in plasma by sparing the consumption of vitamin E under normal physiological circumstances. PMID- 12558056 TI - Dietary flax oil reduces renal injury, oxidized LDL content, and tissue n-6/n-3 FA ratio in experimental polycystic kidney disease. AB - As whole flaxseed is beneficial in the treatment of experimental renal disease, we undertook a study to determine whether previously documented benefits of whole flaxseed could be reproduced with dietary low-lignan flax oil (FO), a rich source of alpha-linolenic acid, in experimental polycystic kidney disease. Male offspring of Han:SPRD-cy heterozygous rats were fed a synthetic diet containing FO or corn oil (CO) for 8 wk from the time of weaning. Renal inflammation, fibrosis, proliferation, cystic change, and oxidized-LDL were assessed morphometrically. Hepatic and renal lipid composition was assessed using GC. FO feeding produced hepatic and renal enrichment of n-3 PUFA and an increase in C18:>C18 PUFA ratios (18-carbon PUFA compared to longer-chain PUFA), with a reduction in proportion of hepatic long-chain PUFA. The FO-based diet was associated with lower mean cystic change by 29.7% (P = 0.018), fibrosis by 21.7% (P = 0.017), macrophage infiltration by 31.5% (P < 0.0001), epithelial proliferation by 18.7% (P = 0.0035), and ox-LDL detection by 31.4% (P < 0.0001) in Han:SPRD-cy heterozygotes. Serum creatinine was significantly lower in FO-fed diseased animals. A small hypocholesterolemic effect was noted in all animals fed FO. FO feeding moderates renal injury, modifies the profile of substrates available for elongation to eicosanoid precursors, and inhibits the elongation of C18 PUFA in this model. The consumption of FO-based products may prove a more practical way of obtaining health benefit than attempts to increase dietary content of unrefined seed. PMID- 12558057 TI - Tuna fishmeal as a source of DHA for n-3 PUFA enrichment of pork, chicken, and eggs. AB - Enriching foods with long-chain n-3 PUFA (LC n-3 PUFA) is an important approach to increasing the dietary intake of these beneficial nutrients. Enrichment of meat and eggs can be achieved by adding flaxseed, fish oil, or fishmeal to pig or poultry feeds. However, utilization of these sources, particularly fishmeal, has been limited by concerns about adverse effects on sensory qualities. In this study, we evaluated the use of PorcOmega (POM), a stabilized tuna fishmeal formulation, as a source of DHA for enrichment of pork and poultry products. Pigs, broilers, and laying hens were fed rations containing varying levels of POM for varying time periods, and its impact on the LC n-3 PUFA content and sensory qualities of cooked meat and eggs was examined. Pork and chicken products, including chops, sausages, and eggs, with substantially increased (up to sevenfold) levels of LC n-3 PUFA (predominantly DHA) were achieved by including up to 10% POM in rations. The increases were retained after cooking. Some sensory deficits were noted when using higher levels of POM (exceeding 15% in meat and 10% in eggs). However, at modest rates of feeding (5-10% POM), palatable meat and eggs were obtained with worthwhile levels of enrichment. The fishmeal feeding strategy used in this study offers a viable means of producing a range of alternative dietary sources of LC n-3 PUFA. PMID- 12558058 TI - Effects of dietary alpha-linolenic acid from blended oils on biochemical indices of coronary heart disease in Indians. AB - PUFA of the n-6 and n-3 series have beneficial effects on key risk factors of coronary heart disease (CHD). Our earlier studies on the intake of FA and on the FA composition of plasma and platelet phospholipids suggested the need to improve the n-3 PUFA nutritional status in the Indian population. The present long-term study was conducted on 80 middle-aged Indian subjects (40 men and 40 women) using the subjects' own home-prepared diets to evaluate the effects of dietary n-3 PUFA on biochemical indices of CHD risk. Substitution of Blend G (equal proportions of groundnut and canola oils) for groundnut oil or substitution of Blend S (equal proportions of sunflower and canola oils) for sunflower oil increased alpha linolenic acid (ALNA) fourfold and decreased the linoleic acid (LA)/ALNA ratio from 35 to 6 and 65 to 9, respectively. Twelve subjects (six men and six women) who received Blend G were switched back to groundnut oil and were administered 0.3 g daily of long-chain (LC) n-3 PUFA from fish oil. At the end of the trial period for both blends in both sexes, plasma lipid and apolipoprotein levels had not changed, and ADP-induced aggregation had decreased. In plasma and platelet phospholipids, LA as well as LCn-3 PUFA had increased, suggesting competition between LA and ALNA for metabolism into the respective LC-PUFA. Fish oil supplementation increased LCn-3 PUFA in plasma and platelet phospholipids, decreased ADP-induced platelet aggregation, and increased plasma cholesterol. On the basis of the increased LCn-3 PUFA in plasma phospholipids, it was calculated that 0.75% energy (en%) (2.2 g) ALNA (from vegetable oils) may be required to increase LCn-3 PUFA to about the same extent as 0.1 en% (0.3 g) LCn-3 PUFA (from fish oils). Since both n-6 and n-3 PUFA play a critical role in fetal growth and development and in the programming of diet-related chronic diseases in adults, an improvement in the n-3 PUFA nutritional status in cereal-based diets through long term use of cooking oils containing 25-40% LA and 4% ALNA may contribute to the prevention of CHD in Indians. PMID- 12558060 TI - Rapid and improved method for the determination of bile acids in human feces using MS. AB - A simple method for the determination of bile acids in adult human fecal samples using GC-MS is described. Bile acids are directly extracted from feces by ethanol (95%) containing 0.1 N NaOH. Extracts are purified by passage through a reversed phase C18 silica cartridge and then analyzed by GC-MS. The present study has shown that lyophilized human feces contain mainly free bile acids, with lithocholic acid (LCA) and deoxycholic acid (DCA) as the major bile acids; however, isomers of LCA and DCA, keto-bile acids, and cholic acid are also present. Any traces of conjugated bile acids are hydrolyzed before the C18 extraction by deconjugating enzymes, which are present in feces and are activated by the addition of water during the homogenization step. Thus, the analysis of fecal bile acids can be performed without the hydrolysis step in less than 4 h in comparison to traditional techniques, which usually require at least 48 h. PMID- 12558059 TI - 31P NMR quantification and monophasic solvent purification of human and bovine lens phospholipids. AB - Most lipid extraction procedures [Folch, J., Lees, M., and Sloane-Stanley, G.H. (1957) A Simple Method for the Isolation and Purification of Total Lipids from Animal Tissues, J. Biol. Chem. 226, 497-509; Bligh, E.G., and Dyer, W.J. (1959) A Rapid Method of Total Lipid Extraction and Purification, Can. J. Biochem. Physiol. 37, 911-917] employ biphasic solvent mixtures designed to dissolve the lipids in an organic phase and remove impurities in an aqueous phase. However, when applying these protocols to biological matrices such as that of the ocular lens, the formation of an emulsion layer between the organic and aqueous phases causes poor reproducibility in extraction yields and gives only a small amount of the lipid-containing chloroform phase. In this study, we quantified phospholipids at each step of the Folch et al. extraction protocol and compared the yield of human and bovine lens phospholipids obtained by the Folch-based approach and a novel monophasic methanol extraction method designed to circumvent the problems associated with biphasic extraction protocols. A monophasic methanol extraction coupled with 31P NMR spectroscopy was found to be the simplest, quickest, and most effective method for quantifying the phospholipid content of the lens. PMID- 12558061 TI - Simultaneous evaluation of HMG-CoA reductase and cholesterol 7alpha-hydroxylase activities by electrospray tandem MS. AB - Simultaneous evaluation of HMG-CoA reductase and cholesterol 7alpha-hydroxylase activities by electrospray tandem mass spectrometry (ES-MS-MS) was performed. The assay was based on the measurement of mevalonolactone (MVL) and 7alpha hydroxycholesterol (7alpha-OHC) produced by the incubation of HMG-CoA with hepatic microsomes in the presence of NADPH and glucose-6-phosphate dehydrogenase. Following extraction and purification using a cyanopropyl cartridge, MVL and 7alpha-OHC were analyzed, without derivatization, by ES-MS-MS. The analysis was achieved in 5 min. Calibration curves were made for MVL and 7alpha-OHC, and were linear from 0 to 100 microg. The recovery was >97%. The procedure was validated under similar calibration and recovery experiments, by measuring the above mentioned products as dimethylethylsilyl ether derivatives using the classical technique of GC-MS. Data obtained by ES-MS-MS and GC-MS showed a good correlation, with no significant differences. ES-MS-MS is a simple and reliable method for the evaluation of HMG-CoA reductase and cholesterol 7alpha-hydroxylase activities in liver microsomal preparations. PMID- 12558062 TI - 5,9,23-Triacontatrienoic methyl ester, an elastase inhibitor from the marine sponge Chondrilla nucula. AB - A polyethylenic fatty ester was isolated from the marine sponge Chondrilla nucula. The structure was elucidated through NMR spectral data and MS analysis as 5,9,23-triacontatrienoic methyl ester 1. Compound 1 is an elastase inhibitor [ID50 = 10 microg/mL (2 x 10(-5) M)]. PMID- 12558063 TI - A critical analysis of new molecular targets and strategies for drug developments in Alzheimer's disease. AB - Alzheimer's disease (AD), a progressive, degenerative disorder of the brain, is believed to be the most common cause of dementia amongst the elderly. AD is characterized by the presence of amyloid deposits and neurofibrillary tangles in the brain of afflicted individuals. AD is associated with a loss of the presynaptic markers of the cholinergic system in the brain areas related to memory and learning. AD appears to have a heterogeneous etiology with a large percentage termed sporadic AD arising from unknown causes and a smaller fraction of early onset familial AD (FAD) caused by mutations in one of several genes, such as the beta-amyloid precursor protein (APP) and presenilins (PS1, PS2). These proteins along with tau, secretases, such as beta-amyloid cleaving enzyme (BACE), and apolipoprotein E play important roles in the pathology of AD. On therapeutic fronts, there is significant research underway in the development of new inhibitors for BACE, PS-1 and gamma-secretase as targets for treatment of AD. There is also a remarkable advancement in understanding the function of cholinesterase (ChE) in the brain and the use of ChE-inhibitors in AD. A new generation of acetyl- and butyryl cholinesterase inhibitors is being studied and tested in human clinical trials for AD. The development of vaccination strategies, anti-inflammatory agents, cholesterol-lowering agents, anti-oxidants and hormone therapy are examples of new approaches for treating or slowing the progression of AD. In addition, nutritional, genetic and environmental factors highlight more effective preventive strategies for AD. Developments of early diagnostic tools and of quantitative markers are critical to better follow the course of the disease and to evaluate different therapeutic strategies. In this review, we attempt to critically examine recent trends in AD research from molecular, genetic to clinical areas. We discuss various neurobiological mechanisms that provide the basis of new targets for AD drug development. All these current research efforts should lead to a deeper understanding of the pathobiochemical processes that occur in the AD brain in order to effectively diagnose and prevent their occurrence. PMID- 12558064 TI - Receptor tyrosine kinases: the main targets for new anticancer therapy. AB - Because conventional chemotherapy is not specific for cancer cells leading to toxic side effects there is a need for novel agents with high grade antitumor specificity. The major prerequisite to develop such drugs is to understand the targets that these agents should attack. In recent years a number of promising new anticancer drugs have been developed which target intracellular pathways or extracellular cell molecules. The clinically most effective compounds function as tyrosine kinase inhibitors. In the past, various tyrosine kinase receptors have been identified as regulators of tumor or tumor vessel growth. Having shown their expression characteristics in different tumor entities, specific inhibitors of the ATP binding sites of these receptors or antibodies were developed and entered clinical trials. The pathognomonic role of the tyrosine kinase defines the way of action of the inhibiting drug, whereas the amount of expression in tumor tissue defines the rationale to use the inhibitor to treat a specific protein. The future will define indications for such drugs by tumor kinase profiles instead of tumor entities. Gleevec, inhibiting the BCR-ABL tyrosine kinase; Iressa, inhibiting the EGF-receptor tyrosine kinase; Herceptin, inhibiting the Her2/neu tyrosine kinase and PTK787/ZK222584, inhibiting the VEGF-receptor tyrosine kinase will be discussed as representatives of selective tyrosine kinase inhibitors whereas ZD6474 and SU6668 will be discussed as representatives of multitarget tyrosine kinase inhibitors. PMID- 12558065 TI - Integrin alpha v beta 3 as a therapeutic target for blocking tumor-induced angiogenesis. AB - The integrin receptor alphavbeta3 has been shown to play a critical role in several distinct processes, such as angiogenesis, osteoclast-mediated bone resorption and tumor metastasis. Its expression is upregulated in newly synthesized blood vessels produced in response to a variety of tumors and purified angiogenic factors. Studies show that alphavbeta3 is a critical target downstream from perhaps all angiogcnic factors. Proof-of-principle that alphavbeta3 antagonists such as monoclonal antibodies and small molecules block angiogenesis and tumor growth has been obtained in several animal models. Many endogenous inhibitors of angiogenesis such as angiostatin, endostatin and tumstatin seem to work through the alphavbeta3 receptor further emphasizing the critical role of this receptor in angiogenesis. In addition, the alphavbeta3 receptor has been clearly implicated in several pathological processes such as rheumatoid arthritis, osteoporosis, and metastasis of prostate cancer to bone. Thus alphavbeta3 may prove to be an important target for pharmacological intervention in more than one clinical setting. PMID- 12558066 TI - 3,7-bis(dialkylamino)phenothiazin-5-ium derivatives: biomedical applications and biological activity. AB - The light-induced reactions of 3,7-bis(dialkylamino)phenothiazin-5-ium compounds with biological substrates are briefly discussed. Their biomedical applications, in particular those related with biological staining, interaction with proteins and antiviral, antibacterial and antitumour activity are reviewed. PMID- 12558067 TI - Regulation of gene expression in vascular cells by coagulation proteins. AB - Receptors of vascular cells and coagulation proteins form a tightly integrated and balanced system, providing regulation to coagulation and mediating a response to coagulation by the vascular cells. Endothelial and smooth muscle cells express a variety of proteins directly participating in hemostasis. Engagement of activated coagulation proteins by their specific receptors on the vascular cell surface, in turn, activates these cells and leads to expression of genes involved in coagulation, angiogenesis, leukocyte adhesion, regulation of the vascular wall tone, etc. The signals inducing the expression of target genes are mediated by protease-activated receptors, which are shared among coagulation proteases. However, differences in mechanisms of activation of these receptors, as well as the presence of specific receptors for each coagulation protein and structures of promoters of target genes may provide specificity in the responses of vascular cell types to different coagulation factors. Activation of gene expression in vascular cells by coagulation proteases accounts for the long-term consequences of coagulation in disorders such as atherosclerotic lesion development, cancer growth, and inflammation. Multiple intracellular pathways and specific transcriptional mechanisms activated by coagulation proteins represent an attractive target for drug design, providing the possibility of controlling the adverse effects of coagulation activation without interfering with the hemostatic requirements of coagulation. This review discusses regulation of gene expression in vascular cells by thrombin, tissue factor, factor VIIa, factor Xa and protein C. Differences and similarities in mechanisms of receptor activation, the pathological profiles of genes activated by these coagulation factors, and recently described transcriptional mechanisms that they induce are discussed. PMID- 12558068 TI - Current targets for anticancer drug discovery. AB - The call for the discovery of less toxic, more selective, and more effective agents to treat cancer has become more urgent. Inhibition of angiogenesis continues to be one of the main streams in the current cancer drug discovery activity. Insights into tumor angiogenesis biology have led to the identification of a number of molecules, which are important for the progression of these processes. Of particular interest is a group of growth factors including fibroblast growth factor, platelet-derived growth factor, and vascular endothelial growth factor. These growth factors and their corresponding receptor tyrosine kinases have become important targets for inhibition of the proliferation of endothelial cells, the main component of blood vessels. The validated targets for inhibition of angiogenesis also include a family of matrix metalloproteinases and cell adhesion molecules. In the closely related area, protein kinases have emerged as one of the most important targets for drug discovery. Besides growth factor receptor tyrosine kinases, numerous other protein kinases implicated in malignancies have been identified including non receptor kinases such as Bcl-Abl and Src kinases. In addition, the cell cycle regulators (cyclin-dependent kinases, p21 gene) and apoptosis modulators (Bcl-2 oncoprotein, p53 tumor suppressor gene, survivin protein, etc) have also attracted renewed interest as potential targets for anticancer drug discovery. Other molecular targets include protein farnesyltransferase (FTase), histone deacetylase (HDAC), and telomerase, which have essential roles in cellular signal transduction pathways (FTase, HDAC) and cell life-span (telomerase). This review presents a comprehensive summary and discussion on the most important targets currently attracting a great deal of interest in contemporary anticancer drug design and discovery. Recent advances complementing these targets are also highlighted. PMID- 12558069 TI - Fluoroquinolones: structure and target sites. AB - The quinolones are a potent group of drugs that target the essential bacterial enzymes DNA gyrase and topoisomerase IV. DNA gyrase is the primary target of Gram negative organisms however, it is topoisomerase IV that is the primary target of Gram positive organisms. Within these enzymes is a highly conserved region centered round the active site where resistance mutations occur. These mutations are almost always identical, irrespective of organism. In spite of the homology of this region, amino acid sequence analysis shows that there are defined differences between the Gram groups, particularly in topoisomerase IV, and it is speculated that herein lies the origin of target preference. Since the first quinolone nalidixic acid was developed, the quinolones have undergone structural modifications, in particular the addition of a fluorine at position 6, to produce the fluoroquinolones. This has seen their potency and pharmakokinetic profile greatly increase. In vitro selection of resistance mutations has allowed the observation of how resistance is acquired and some of the modifications in newer fluoroquinolones have resulted in the shift of primary target from topoisomerase IV to gyrase with Gram positives. Curiously, purified topoisomerase IV is still more sensitive even if gyrase is the primary target. Gyrase remains the primary target for Gram negatives. PMID- 12558070 TI - The study of HLA class II and autoimmune diabetes. AB - Many autoimmune diseases have genetic associations with the Major Histocompatibility Complex (MHC) class II loci. Susceptibility to Type 1 diabetes mellitus (TIDM) is particularly associated with Human Leucocyte Antigen (HLA) DR3, 4 and associated DQ2, 8 alleles and this is well documented in genetic association studies. These molecules play an important role in presentation of peptide antigens after intracellular processing to CD4 T lymphocytes. During the last decade, a number of approaches have been used to elucidate the molecular basis for the association of particular alleles with susceptibility to or protection from TIDM. These studies have focused on investigating the structure of the antigen presenting molecules, together with their peptides. Through binding studies, peptide elution, molecular modelling and crystallization of the peptide MHC complex, it has been possible to define the peptide binding regions and examine the stability of binding of peptides from putative autoantigens. This knowledge has also facilitated the development of reagents such as multimeric MHC peptide complexes that will help to track the low frequency, potentially pathogenic antigen specific cells. Recently, HLA transgenic mice have been generated and used to study T cell epitopes. In addition, although it is clear that the presence of HLA molecules alone does not by itself cause disease, these transgenic mice will develop diabetes when there is an islet "insult", even if the islet "insult" is, itself, not sufficient to precipitate disease in the absence of the HLA class II transgene. These mice will allow further study of the role of these HLA molecules in vivo. We now have a much greater general understanding of the possible reasons why particular molecules may encode susceptibility to or protection from disease. All these studies will provide information to ultimately define a rational basis for the development of targeted immunotherapy. PMID- 12558071 TI - Recent advances on alpha-synuclein cell biology: functions and dysfunctions. AB - Alpha-synuclein is a recently discovered protein that was first identified as the major non amyloid component of senile plaques, the cerebral lesion likely responsible for Alzheimer's disease. The role of alpha-synuclein in another brain disease namely Parkinson's disease, has been more deeply documented. It appears that alpha-synuclein fills up the intracytoplasmic inclusions called Lewy bodies that likely contribute to the etiology of Parkinson's disease. Furthermore, rare familial forms of Parkinson's disease have been shown to be linked to autosomal dominant mutations of alpha-synucleins. Is alpha-synuclein a bridge between Alzheimer's and Parkinson's diseases? Could it be seen as a common denominator for these two neurodegenerative diseases? These issues could be better addressed by further delineating the physiological function of alpha-synuclein and, as a corollary, the dysfunction taking place along with the diseases. Here, I will review the recent advances concerning the physiology of alpha-synuclein and will particularly focus on the post-traductional events leading to drastic biophysical transformations. I will describe recent works suggesting that these modifications directly modulate the normal function of alpha-synuclein, likely accounting for the dysfunction associated with Parkinson's disease and perhaps contributing to Alzheimer's pathology. PMID- 12558072 TI - The posttranslational phase of gene expression: new possibilities in molecular diagnosis. AB - Proteins in general and secretory proteins in particular undergo posttranslational processes before they reach the structure in which they can fulfill their functional purpose. The protein precursor may undergo a wide variety of proteolytic cleavages, N- and C-terminal trimmings and amino acid derivatizations in cells that express the protein. Occasionally, the same precursor is differently processed in different cell types and, in addition, diseased cells may process a given precursor abnormally. For instance, the translational process is often either increased or decreased in diseased cells, which render the ensuing modifications of the precursor incomplete. As a result, a variable mixture of precursors and processing-intermediates accumulates. Measurement of a single protein or peptide component of the posttranslational processing cascade may not facilitate the diagnosis of a disease, because the pattern of precursors and processing products vary individually among patients. In order to exploit disturbed posttranslational processing for diagnostic use, and--at the same time--provide an accurate measure of the translational product, a simple analytical principle named "processing-independent analysis" (PIA) has been designed. PIA-methods quantitate the total mRNA product irrespective of the degree of precursor processing. PIA-methods have recently been developed for a number of prohormones and neuroendocrine proteins, and their diagnostic potential appears promising in early diagnosis of tumors and cardiovascular diseases. The present review describes posttranslational processing patterns for some neuroendocrine proteins. Second, PIA-measurements of precursor-products are mentioned with indication of problems and pitfalls. Finally, PIA-results obtained in diagnosis of neoplastic and cardiovascular diseases are highlighted. But first general aspects of the posttranslational processing are reviewed as a necessary basis for the understanding of the new diagnostic possibilities. PMID- 12558073 TI - Cytokine polymorphisms in chronic inflammatory diseases with reference to occupational diseases. AB - Genes which encode inflammatory cytokines are subject to polymorphisms in their regulatory regions that may effect both the level and ratio of cytokines produced in response to exogenous stimuli. These variant alleles are observed in a large percent of the population and are often associated with increased or decreased susceptibility or severity (modifiers) to infectious, immune or inflammatory diseases. Environmental factors can also play either a direct (i.e., causative factor) or indirect (modifying factor) role in these diseases. Thus, it would follow that gene-environment interactions would effect the expression and/or progression of the disease. In the present review, the concept that some of the common allelic variants found in cytokine genes represent modifying factors in chronic inflammatory diseases associated with occupational exposure is discussed. PMID- 12558074 TI - Understanding the tumor metabolic phenotype in the genomic era. AB - Now, at the beginning of a new century, 80 years after Warburg's Nobel prize winning discoveries, we are beginning to make sense of the underlying causes of the well known metabolic phenotype of tumor cells. Building on decades of research to understand the interrelationships between respiration and glycolysis in cancer, the tumor metabolic phenotype can now begin to be understood in a genomic context. With the discovery of hypoxia inducible factor-1 (HIF-1), which is widely overexpressed across a broad range of cancers, modern molecular tools have allowed us to put together the pattern of events that might explain the metabolic differences between tumor and normal cells. HIF-1 controls cellular and systemic responses to oxygen availability and coordinates up-regulation of genes involved in many pathways concerned with tumour growth and metabolism including angiogenesis, glucose and energy metabolism, cellular proliferation, differentiation and viability, apoptosis, pH regulation and matrix metabolism. These findings begin to explain how glucose uptake and glycolysis could be up regulated in cancer cells (through binding to a core DNA recognition sequence) in a co-ordinated and constitutive fashion that may also allow us to elucidate new targets for tumor therapy. PMID- 12558075 TI - Novel approaches to cancer therapy using oncolytic viruses. AB - The goal of oncolytic therapy is to exploit the innate ability of viruses to infect tumor cells, replicate in tumor cells, and produce selective oncolysis while sparing normal cells. Although the concept that viruses can be oncolytic is not new, it is only in the last three decades that efforts have been directed at genetically mutating viruses to specifically target characteristics of cancer cells. Several viruses have the potential to infect, replicate and lyse tumor cells, each taking advantage of different host cancer cell biology. This review will focus on the major viruses under current investigation for oncolytic therapy, the mechanism by which they specifically eradicate tumors, and the clinical strategies currently under investigation. PMID- 12558076 TI - Signaling pathways regulating gliomagenesis. AB - The astrocytomas represent the most common primary tumors of the brain. Despite efforts to improve the treatment of astrocytomas, these tumors and in particular the high-grade astrocytoma termed glioblastoma multiforme still carry a poor prognosis. In recent years, there has been an intensive effort to gain an understanding of the cellular and molecular mechanisms that contribute to the pathogenesis of astrocytomas as a first step toward the development of better treatments for these devastating tumors. Here, we will review our current understanding of the signaling pathways that underlie glial transformation. Studies of astrocytomas have led to the identification of two major groups of signaling proteins whose abnormalities contribute to gliomagenesis: the cell cycle pathways and the growth factor-regulated signaling pathways. Among the cell cycle proteins, the p16-cdk4-pRb and ARF-MDM2-p53 cell cycle arrest pathways play a prominent role in glial transformation. In addition, deregulation of polypeptide growth factors acting via receptor tyrosine kinases (RTKs) and of intracellular signals, including the lipid phosphatase PTEN, that regulate cellular responses to RTKs plays a critical role in gliomagenesis. In addition to the identification of the signaling proteins targeted in glial transformation, the cell-of-origin of astrocytomas has been investigated. Genetic modeling of astrocytomas in mice suggests that neuroepithelial precursor cells represent preferred cellular substrates of gliomas or that either astrocytes or precursor cells constitute potential cells-of-origin of astrocytomas. During normal brain development, neuroepithelial precursor cells, including neural stem cells, differentiate into astrocytes. As the mechanisms that control gliogenesis during normal brain development become better understood, it will be important to determine if deregulation of these mechanisms might contribute to the pathogenesis of astrocytomas. The elucidation of the molecular underpinnings of astrocytomas holds the promise of improved treatment options for patients with these devastating brain tumors. PMID- 12558077 TI - Protein-tyrosine kinases and adaptor proteins in FcepsilonRI-mediated signaling in mast cells. AB - Mast cells function as the initiator of the allergic reaction and play a role in the innate immune system. Aggregation of the high affinity IgE receptor (FcepsilonRI) on mast cells triggers degranulation with the release of chemical mediators such as histamine, production of cytokines and leukotrienes. FcepsilonRI signals by activating proximal non-receptor type of protein-tyrosine kinases, Lyn, Syk, Btk and Fyn. Activated tyrosine kinases then phosphorylate their specific substrates which include other enzymes and adaptor proteins and assemble these cytoplasmic signaling molecules for cellular activation. The adaptor proteins have multiple domains that allow binding to effector molecules and therefore act as positive or negative regulators controlling FcepsilonRI signaling. Deletion of the adaptor proteins such as LAT, SLP-76 or Gab2 resulted in decreased FcepsilonRI-mediated anaphylactic reaction in vivo. Functional analysis of adaptor proteins has raised the possibility that they may be new targets for the discovery of anti-allergic drugs. PMID- 12558078 TI - Abdominal incisions: techniques and postoperative complications. AB - BACKGROUND AND AIMS: The choice of incision for laparotomy depends on the area that needs to be exposed, the elective or emergency nature of the operation and personal preference. Type of incision may however have its influence on the occurrence of postoperative wound complications. Techniques and features of various incisions are discussed, as well as the incidence of their postoperative complications. METHOD: A medline search was conducted identifying prospective randomised trials, as well as retrospective studies with sufficient follow-up, comparing midline, paramedian, transverse and oblique incisions. RESULTS: Significant differences in wound infection and wound dehiscence rates were not reported. Transverse, oblique and paramedian incisions caused significantly less incisional hernias than midline incisions. However, trials comparing transverse and midline incisions for larger laparotomies did not show significant differences. All four trials comparing lateral paramedian with midline incisions reported incisional hernia rates of 0% after the lateral paramedian incision. Differences with the midline incision were significant. CONCLUSION: Transverse or oblique incisions should be preferred for small unilateral operations. The paramedian incision should be used for major elective laparotomies. The use of the midline incision should be restricted to operations in which unlimited access to the abdominal cavity is useful or necessary. PMID- 12558079 TI - CT- and MR colonography. AB - BACKGROUND: Colorectal cancer is the second most frequent cancer and adenomas are widely accepted as precursors to colorectal cancer. Diagnosis and removal of adenomas are recommended to reduce cancer incidence and mortality. The current diagnostic methods include sigmoidoscopy and colonoscopy. Lately, CT- and MR colonography have emerged as non-invasive methods for colon imaging. METHODS: At present, CTC and MRC require bowel preparation. However, preliminary studies have been carried out without colon preparation. After the colon has been filled with air or contrast, the patient is scanned in the supine and prone positions. Data are then downloaded to a workstation for post processing and image-analysis. RESULTS: Results have shown a high sensitivity and specificity for polyps > or = 10 mm, comparable to the sensitivity of conventional colonoscopy and superior to double contrast barium enema. CONCLUSIONS: With the exponential development in computer processing power, CT- and MR colonography holds the promise for future colon examination with the advantages of non-invasiveness, no need for sedation, and probably no bowel preparation. A major disadvantage, however, is the radiation dose during CT colonography. Future developments with the use of "intelligent" computers, better resolution and faster examinations will make CT and/or MR colonography realistic options to replace conventional diagnostic colonoscopy. PMID- 12558080 TI - Fatal firearm injuries in Finland: a nationwide survey. AB - BACKGROUND AND AIMS: Treatment of life-threatening firearm injuries represents major challenges to the involved medical staff. The aim of the study was to assess numbers, natures and injury patterns of fatal incidents by firearms in Finland over a 10-year period. MATERIAL AND METHODS: Retrospective analysis of all firearm-related deaths during the 10-year period from January 1990 to December 1999. Death certificates were obtained and reviewed with detailed analysis of non-suicidal deaths. RESULTS: Over the 10-year period 1990 to 1999, 452 individuals (equivalent to 1.8 cases per 100 000 person-years) died in Finland as a result of accidental or violent shooting. There were no significant changes in numbers of cases from year to year. The male-female ratio was 7:3. The mean age of the victims was 32 years. The shooting incident had been classified as an assault in 78% of cases. Its nature was unspecified in 13% of cases and clearly accidental in 9%. The predominant anatomical site of fatal injury following assault was the chest in 43%, the head in 42% and the abdomen in 8% of the cases. After accidental or unspecified nature shooting, the predominant site of injury was the head in 68%, followed by the chest in 16% and the abdomen in 10% of the cases. Most victims (86%) died at the scene of the shooting. Only 14% were alive at hospital admittance. Most hospitalised victims died within 24 hours of admission. CONCLUSIONS: The annual incidence of fatal non-suicidal firearm injuries did not change significantly in Finland between 1990 and 1999. Victims of such injuries required only minimal hospital resources because in most cases they died at the scene of the shooting. The results of the study reported indicate that efforts to prevent fatal injuries from use of firearms or diminish their number should be focused mainly on prevention of firearm related assault. PMID- 12558081 TI - Radioisotope-guided surgical biopsy of costal metastases in breast cancer patients. AB - Radioisotope bone scanning is a sensitive indicator of metastatic disease. Localization of the hot spot for biopsy is difficult. Today, a hand-held gamma probe enables the surgeon intraoperatively to accomplish the localization procedure easily. As this technique is described in American literature exclusively, more recent European findings is presented here. During one month two patients were diagnosed with breast cancer. Bone scanning disclosed minor hot spots in the ribs. On the day of surgical biopsy the localization of the hot spots was guided by a gamma probe. For the biopsy a dermal punch biopsy needle was employed, and fine needle aspiration biopsies were also taken. The procedures were easy and completed within a few minutes. The post-operative courses were uneventful. In one patient, final histology showed malignancy in both biopsies. In the other patient, only the fine needle aspiration biopsy was malignant. The punch biopsy was abnormal, but did not contain malignant tissue. The gamma probe is an important tool in the performance of rib biopsies of nonpalpable lesions. The punch biopsy technique is simple and combined with fine needle aspiration biopsy the method has proven to be sensitive and accurate. The punch biopsy technique furthermore reduces the risk of postoperative pneumothorax. PMID- 12558082 TI - Inguinal sentinel lymph node biopsy for staging anal cancer. AB - BACKGROUND AND AIMS: The optimal treatment of clinically negative inguinal lymph nodes in patients with primary anal cancer has not yet been clearly defined. The presence of metastases in the inguinal lymph nodes is an adverse prognostic factor for anal cancer. In the present study the feasibility of sentinel lymph node biopsy (SLNB) for staging anal cancer was investigated. PATIENTS AND METHODS: From September 1999 to March 2002, 8 patients with biopsy proven primary anal cancer underwent lymphoscintigraphy and dual-agent guided inguinal SLNB for nodal staging before starting multimodality treatment. RESULTS: Inguinal SLNB was successful in all 8 patients (13 groins). A total of 20 hot and blue SLNs (mean 1,5 (1-2) per groins) were removed. In 2 patients (25%) the SLN was positive for metastasis. CONCLUSIONS: Lymphoscintigraphy followed by dual-agent guided inguinal SLNB is technically feasible for staging patients with primary anal cancer. The detection of metastases in the removed sentinel lymph node(s) may alter the treatment and thus may improve the locoregional control and overall survival of these patients. PMID- 12558083 TI - Feasibility of predicting the risk of atrial fibrillation after coronary artery bypass surgery with logistic regression model. AB - BACKGROUND AND AIMS: The aim of this study was to determine the risk factors of postoperative atrial fibrillation (AF) after coronary artery bypass grafting and to create predictive model and to evaluate the effects of AF on patients outcome. MATERIAL AND METHODS: Data of 3,676 consecutive patients were analysed to identify the predictors of AF. Multivariate logistic regression model was validated prospectively in 1,107 patients. RESULTS: Increasing age (p < 0.001), preoperative use of digoxin (p = 003), need of intra-aortic balloon pump or inotropic medication in the weaning off cardiopulmonary by pass or during the first 24 hours postoperatively (p = 0.013), increasing body surface area (p = 0.006) and lower ejection fraction (p = 0.048) were independent risk factors for postoperative AF. The predictive model gave area under the receiver-operating characteristic (ROC) curve 0.682, 95% confidence interval 0.663-0.701, and p < 0.001. The patients with AF incidence had more postoperative stroke (p = 0.008), confusion (p < 0.001) severe gastrointestinal complications (p = 0.005), readmission to ICU (p < 0.001), longer ICU (p < 0.001) and hospital stay (p < 0.001) when compared with the patients who remained in sinus rhythm. CONCLUSION: Logistic regression model with the parameters used was not accurate enough for clinical purposes. Postoperative AF is associated with postoperative stroke, severe gastrointestinal complications, readmission to ICU, and longer ICU and hospital stay. PMID- 12558084 TI - The fate of AAA patients referred electively to vascular surgical unit. AB - BACKGROUND: The ideal treatment of abdominal aortic aneurysms (AAA) is to operate aneurysms likely to rupture, without exposing other cases to major surgery. The purpose here was to analyse retrospectively the management of AAA in a well defined geographical region in the 1990's. METHODS: 194 new vascular surgical outpatient consultations due to AAA were done to the regional vascular centre during the years 1990, 1992, 1994, 1996 and 1998. Data were collected from case records. Statistics Finland provided causes and dates of death. RESULTS: The mean observed annual AAA incidence was 9.0 per 100 000 inhabitants and it rose significantly (33.3%) during the study period. The duration of follow-up varied between 0 and 129 months. The 5/8-year cumulative mortality was 37.3/50.7%. The most common causes of death were AAA-related (31.7%), cardiac (29.1%) or malignancy (19.0%). Twenty-five patients with small AAA were referred to primary health care sector for further follow-up. There were no RAAA (ruptured AAA) deaths in this group. The cumulative 5/8-year mortality was 43.2/49.9%. One hundred patients underwent an elective aneurysm repair with in-hospital mortality of 7.0%. The cumulative 5/8-year mortality was 23.7/35.4%. Twelve patients refused elective treatment. The cumulative 5/8-year mortality was 45.1/ 63.4% and 5/7 deaths were due to RAAA. Twenty-three patients were unfit for elective repair. The cumulative 5/8-year mortality was 87.0%/100% and 5/20 deaths were caused by RAAA. The cumulative 5/8-year RAAA-rate in the patients with AAA more than 5.0 cm in diameter and outside elective aneurysm-repair (n = 23) was 51.9%/100.0%. CONCLUSION: The observed incidence of AAA increased during the 1990's. Half of the patients underwent an elective procedure. Patients unfit for surgery died mainly for other reasons than RAAA. Most patients with AAA over 5.5 cm not subjected to elective procedure, died of rupture. PMID- 12558085 TI - Do normal leucocyte count and C-reactive protein on admission to hospital exclude a life-threatening attack of acute pancreatitis? AB - BACKGROUND AND AIMS: The early severity assessment of an attack of acute pancreatitis is clinically of utmost importance. The aim of the present work was to study the role of leucocyte count and C-reactive protein (CRP) measurements on admission to hospital in assessing the severity of an attack of acute pancreatitis. In particular, patients with a life-threatening attack of acute pancreatitis but a normal leucocyte count and CRP level were sought. MATERIAL AND METHODS: A total of 1050 attacks of acute pancreatitis were treated at Turku University Central Hospital during the years 1995-1999. Leucocyte count and C reactive protein (CRP) value were determined on admission to hospital. There were 58 life-threatening attacks of acute pancreatitis (group A). Fifty-eight consecutive mild attacks served as controls (group B). The number of patients with both values normal, only leucocyte count raised, only CRP level raised and both values raised were calculated in the groups A and B. RESULTS: Both leucocyte count and CRP level were significantly (P < 0.001 in both comparisons) higher on admission to hospital in patients with a life-threatening disease (group A) than in those with a mild disease (group B). Group A contained no patients with both values in the normal range. In group B, one fifth of the patients had both values normal. CONCLUSION: It is very unlikely that acute pancreatitis proves to be a life-threatening one when both the leucocyte count and CRP are normal on admission to hospital. In the present 1050 acute pancreatitis there were no patients with life-threatening disease but normal laboratory values on admission. PMID- 12558086 TI - The long-term results of Lubinus interplanta hemiarthroplasty in 228 acute femoral neck fractures. A retrospective six-year follow-up. AB - BACKGROUND AND AIMS: We retrospectively studied survival of patients treated with Lubinus interplanta (IP) semiendoprosthesis in acute hip fracture. MATERIALS AND METHODS: The long-term results of Lubinus interplanta (IP) hemiarthroplasty in 228 acute medial fractures of the femoral neck in 222 patients were reviewed. RESULTS AND CONCLUSIONS: The mean age of the patients was 80.3 years. The mean duration of post-operative follow-up was 6.3 years (range 5-8 years). There were 12 reoperations (5.3%), of which 8 were revisions (3.5%). Dislocation rate was 3.5%. Survival was 93% at three years and 88% at six years (Kaplan-Meier-survival analysis). CONCLUSIONS: These results surpass those previously published after Thompson and Moore hemiarthroplasties. The Lubinus prosthesis has a greater CCD (caput,collum, diaphyse) angle and a longer stem compared to Thompson and Moore implants. The need for resection of calcar cortex is also limited. These biomechanical facts may explain the good long-term results of Lubinus hemiarthroplasty. PMID- 12558087 TI - Acute traumatic hemarthrosis of the knee. Is routine arthroscopic examination necessary? A study of 320 consecutive patients. AB - BACKGROUND AND AIMS: Distortions and contusions of the knee are common. Fairly often they will result in hemarthrosis. The purpose of this study was to evaluate the findings in patients with acute traumatic hemarthrosis of the knee and based on these findings estimate the necessity of acute arthroscopic examination. MATERIAL AND METHODS: A total of 320 patients were examined arthroscopically between the years 1994 to 96 in the Turku University Hospital. Patients with other than chondral or osteochondral fractures were excluded. RESULTS: The most common arthroscopic findings were rupture of the ACL (45%), dislocation of the patella (23%) and meniscal tear (21%). In only 113 (35%) cases an immediate therapeutic procedure was performed in addition to the arthroscopic examination. CONCLUSIONS: Based on our findings we believe that routine arthroscopic examination is not necessary in patients with acute traumatic hemarthrosis of the knee. In our opinion the patient should be examined and followed by an orthopedic surgeon and if a lesion requiring operative treatment is diagnosed or suspected, an arthroscopic examination should be scheduled. The timing of the procedure should be such that all possible lesions encountered can be treated in the same operation. PMID- 12558088 TI - Peroral tramadol premedication increases postoperative nausea and delays home readiness in day-case knee arthroscopy patients. AB - BACKGROUND AND AIMS: To evaluate the effect of preoperative oral tramadol on postoperative pain and its effect on the patient's home-readiness after diagnostic day-case knee arthroscopy performed under spinal anaesthesia. MATERIAL AND METHODS: We studied 156 outpatients in a prospective, randomized, double blind fashion to examine the postoperative analgesic effect of preoperative oral slow-release tramadol. Postoperative pain was measured using a 100-mm visual analogue scale (VAS). Postoperative nausea and vomiting (PONV) and the patients home-readiness were assessed. RESULTS: There were no statistically significant differences in the postoperative VAS scores between the tramadol and placebo groups, nor was there any significant difference in the need for postoperative pain medication. Patients in the tramadol group had higher incidence of PONV and they were discharged from hospital later than those given placebo although the tramadol patients required less intravenous midazolam for sedation. CONCLUSIONS: Preoperatively given slow-release tramadol is ineffective for reduction of postoperative pain after day-case arthroscopy of the knee. Additionally, preoperative tramadol is associated with higher incidence of PONV and it seems to cause delay in the patient's home-readiness. PMID- 12558089 TI - Good reduction of paresthesia and pain after excision of excessive callus formation in patients with malunited clavicular fractures. AB - BACKGROUND AND AIMS: Sequelae following non-surgical treatment of clavicular fractures are not uncommon. The aim with this study was to describe results after excision of excessive callus in patients with late onset of paresthesia/numbness. MATERIAL AND METHODS: Eight patients with hypertrophic callus after a malunited clavicular fracture were operated with removal of excessive callus and scar tissue, without stabilizing ostheosynthesis. The reason for surgery was paresthesia/numbness in the arm and hand. There was a delay between trauma and onset of symptoms on average 10 (6-18) months. Neurophysiology prior to surgery was normal in the patients tested. The mean age at trauma was 29 years and the mean time between trauma and operation was 66 months. A prerequisite for surgery was symptoms without recovery for at least 6 months. RESULTS: At follow-up on average 20 months after surgery the paresthesia/numbness, pain at rest, pain during activity, headache and dizziness were significantly reduced or even gone in all patients. The improvement occurred almost immediately after surgery with no signs of recurrence during follow-up. CONCLUSION: Removal of excessive callus and scar tissue after a malunited clavicular fracture can reduce or fully relieve disabling paresthesia and pain even if surgery is done several years after the fracture. Neurophysiological examination is normal due to lack of permanent neurologic damage and can not be used to confirm the symptoms. PMID- 12558090 TI - Chronically homeless women's perceived deterrents to contraception. AB - CONTEXT: Despite the growing number of homeless women of reproductive age, little is known about their reproductive health and contraceptive use. METHODS: A representative sample of 974 currently homeless women surveyed in Los Angeles County in 1997 included 229 who were chronically homeless and at risk for unintended pregnancy. The relationships between perceived deterrents among these women and their frequency of contraceptive use were evaluated using chi-square tests. Logistic regression analyses were performed to identify independent predictors of contraceptive deterrents. RESULTS: The most commonly cited deterrents to contraceptive use were side effects, fear of potential health risks, partner's dislike of contraception and cost (20-27%). Women who reported substantial deterrents were significantly less likely than others to use contraceptives consistently. Perceived deterrents differed by women's characteristics: Hispanic women were more likely than whites to view not knowing how to use contraceptives or which method to use as a deterrent (odds ratios, 9.6 9.8); black women were more likely than whites to cite not knowing which method to use (4.0), lack of storage (5.5), health risks (6.0) and discomfort (3.3). Women with a history of drug abuse had elevated odds of citing not knowing how to use contraceptives, uncertainty about which method to use and cost as deterrents (5.2-7.7). CONCLUSION: Homeless women report substantial deterrents that reduce their contraceptive use. The multiplicity and heterogeneity of their perceived deterrents suggest a need for reproductive health services that are more integrated and culturally competent than those currently offered. PMID- 12558091 TI - Marriage among unwed mothers: whites, blacks and Hispanics compared. AB - CONTEXT: Much of the debate over welfare reauthorization centers on whether marriage promotion should play a key role. Few studies, however, have tracked the marriage and divorce histories of unwed mothers, including minority women, who are often the main targets of welfare reform. METHODS: Data from the 1995 National Survey of Family Growth were used to estimate the hazards of the transition to marriage for women who delayed childbearing until marriage and for teenagers and older women who had a nonmarital first birth, and of the transition to divorce among the ever-married. Life-table estimates calculated with these estimated transition hazards show the cumulative proportions married and divorced, by race and ethnicity, for women who had a nonmarital first birth and for those who did not. RESULTS Nonmarital childbearing reduces the likelihood of marriage. Some 82% of white women, 62% of Hispanics and 59% of blacks who had a nonmarital first birth had married by age 40; the corresponding proportions among those who avoided nonmarital childbearing were 89%, 93% and 76%, respectively. There is no evidence to suggest that the negative effect of nonmarital childbearing on marriage is caused by other observed or unobserved differences between unwed mothers and women who remain childless until marriage. Nonmarital childbearing raises the likelihood of divorce among unwed mothers who eventually marry, a finding that also varies by race and ethnicity. CONCLUSIONS: Marriage promotion policies should focus on lowering rates of nonmarital childbearing. Reductions in nonmarital childbearing, however, may not eliminate long-standing discrepancies in marriage rates between black and white women. PMID- 12558092 TI - Contraceptive use among U.S. women having abortions in 2000-2001. AB - CONTEXT: Knowing the extent to which contraceptive nonuse, incorrect or inconsistent use, and method failure account for unintended pregnancies ending in abortion, as well as reasons for nonuse and imperfect use, can help policymakers and family planning providers support effective contraceptive use. METHODS: Contraceptive use patterns among a nationally representative sample of 10,683 women receiving abortion services in 2000-2001 were examined, as well as reasons for nonuse, problems with the most frequently used methods and the impact emergency contraceptive pills have had on abortion rates. RESULTS: Forty-six percent of women had not used a contraceptive method in the month they conceived, mainly because of perceived low risk of pregnancy and concerns about contraception (cited by 33% and 32% of nonusers respectively). The male condom was the most commonly reported method among all women (28%), followed by the pill (14%). Inconsistent method use was the main cause of pregnancy for 49% of condom users and 76% of pill users; 42% of condom users cited condom breakage or slippage as a reason for pregnancy. Substantial proportions of pill and condom users indicated perfect method use (13-14%). As many as 51,000 abortions were averted by use of emergency contraceptive pills in 2000. CONCLUSIONS: Women and men need accurate information about fertility cycles and about the risk of pregnancy when a contraceptive is not used or is used imperfectly. Increased use of emergency contraceptive pills could further reduce levels of unintended pregnancy and abortion. PMID- 12558093 TI - Sexual intercourse and the age difference between adolescent females and their romantic partners. AB - CONTEXT: The age difference between a female and her partner may influence relationship dynamics in ways that put the female at increased risk of unintended pregnancies and sexually transmitted diseases. Very little is known, however, about how romantic involvement progresses to intercourse, particularly among adolescent females with older male partners. METHODS: Data from 1,975 female participants in the National Longitudinal Study of Adolescent Health were analyzed using logistic regression to determine whether the age difference between an adolescent female and her romantic partner is a risk factor for sexual intercourse. RESULTS: Adolescent females involved with an older partner have higher odds of having intercourse with that partner than females with partners their own age, after adjustment for demographic covariates. The magnitude of this association is most dramatic among the youngest females--for example, the odds of intercourse among 13-year-old females with a partner six years older are more than six times the odds among 13-year-old females with a same-age partner (odds ratio, 6.4), while 17-year-old females with partners six years older have about twice the odds of intercourse when compared with those who have a same-age partner (2.1). CONCLUSIONS: Young adolescent females with substantially older partners are much more likely than their peers to have sex with their partner, which exposes them to the risks of pregnancy and sexually transmitted diseases. PMID- 12558094 TI - Current contraceptive research. PMID- 12558095 TI - Parameterisation, evaluation and comparison of pesticide leaching models to data from a Bologna field site, Italy. AB - Effective prediction of pesticide fate using mathematical models requires good process descriptions in the models and good choice of parameter values by the user. This paper examines the ability of seven pesticide leaching models (LEACHP, MACRO, PELMO, PESTLA, PLM, PRZM and VARLEACH) to describe an arable field environment where sunflowers are grown in the Po Valley, northern Italy. Two pesticides were considered, aclonifen and ethoprophos. The models were evaluated in terms of their ability to reproduce field data of soil water content and pesticide residues in the soil and ground water. The evaluation was based on a combination of calibrated and uncalibrated runs. The results from the models were compared with each other to explore the differences between the models. The models varied in their ability to predict soil water content in the summer: the capacity models PRZM, PELMO and VARLEACH predicted less drying than MACRO, PESTLA, PLM and LEACHP. The models varied in their ability to simulate the persistence of the pesticides in the soil. Differences in the simulated pesticide degradation rate were observed between the models, due to variations in the simulated soil water content and soil temperature, and also differences in the equation linking degradation rate to soil water content. There were large differences among the predictions of the models for the mean leaching depth of ethoprophos. PRZM, PELMO, PESTLA and LEACHP all showed similar mean leaching depth to each other, whereas VARLEACH predicted lower ethoprophos mobility and PLM and MACRO predicted greater mobility. All the models overpredicted dispersion of ethoprophos through the soil profile, as compared to the field data. None of the models was able to simulate the field data of rapid leaching of pesticide to ground water except PLM after calibration of the percentage of macropores in the mobile pore space. More work is required in the parameterisation of macropore flow for those models that include this process. PMID- 12558096 TI - Isolation of a Pseudomonas fluorescens metabolite/exotoxin active against both larvae and pupae of vector mosquitoes. AB - A formulation was developed from the metabolite(s) of a novel Pseudomonas fluorescens Migula strain (VCRC B426) and tested against 4th-instar larvae and pupae of three species of vector mosquitoes, Anopheles stephensi Liston, Culex quinquefasciatus Say and Aedes aegypti (L). The larvae and pupae of An. stephensi were the most susceptible to the formulation, followed by those of C. quinquefasciatus and Ae. aegypti, in that order, and the dosage requirement for pupal mortality was less than that required for larval mortality. The LC50 dosage requirements for larvae of these mosquito species were, respectively, 70.4, 511.5 and 757.3 microg protein ml(-1), whereas for pupae they were, respectively, 2.0, 9.4 and 19.2 microg protein ml(-1). The lethal fraction was purified from the culture broth and its molecular mass, as determined by high performance liquid chromatography, was 44kDa. This is the first report of a microbial formulation acting upon mosquito pupae, a non-feeding stage. Its mode of action and efficacy to control mosquitoes under field conditions need to be studied further. PMID- 12558097 TI - Synthesis and insecticidal activity of benzoheterocyclic analogues of N'-benzoyl N-(tert-butyl)benzohydrazide: Part 1. Design of benzoheterocyclic analogues. AB - The N'-benzoyl group of N-tert-butyl-N'-benzoyl-3,5-dimethylbenzohydrazide (1) was converted to a series of benzoheterocyclecarbonyl groups in order to investigate the potential usefulness of superimposing a hydrazine insecticide on 20-hydroxyecdysone. A series of analogues with benzodioxole, benzodioxane, benzodioxapine, indole, benzoxazole, benzoxazine or benzothiazole instead of the phenyl group of (1) were synthesized and tested for their insecticidal activity against the common cutworm (Spodoptera litura F). N-tert-Butyl-N'-(3,5 dimethylbenzoyl)-1,3-benzodioxole-5-carbohydrazide and N-tert-butyl-N'-(3,5 dimethylbenzoyl)-2,3-dihydro-1,4-benzodioxine-6-carbohydrazide showed high insecticidal activities, superior to that of (1) and equal to that of the commercial insecticide tebufenozide (RH-5992). PMID- 12558098 TI - Synthesis and insecticidal activity of benzoheterocyclic analogues of N'-benzoyl N-(tert-butyl)benzohydrazide: Part 2. Introduction of substituents on the benzene rings of the benzoheterocycle moiety. AB - A series of N'-benzoheterocyclecarbonyl-N-tert-butyl-3,5-dimethylbenzohydrazide analogues possessing a variety of substituents on the benzene rings of the benzoheterocyle moieties were synthesized and tested for their insecticidal activity. The introduction of a methyl group at the R1 position of the benzoheterocycle moiety strongly increased the insecticidal activity. Among the analogues synthesized, N'-tert-butyl-N'-(3,5-dimethylbenzoyl)-5-methyl-6 chromanecarbohydrazide showed the highest insecticidal activity (LC50 = 0.89 mg litre(-1)). PMID- 12558100 TI - Comparative effects of vetiver oil, nootkatone and disodium octaborate tetrahydrate on Coptotermes formosanus and its symbiotic fauna. AB - The potential of vetiver oil and nootkatone as wood treatments against Coptotermes formosanus Shiraki was examined by assessing the effects on termite tunneling, feeding activity and survival, and the consequences on the symbiont protozoa responsible for wood digestion. Comparisons were made with non-treated wood (control), wood treated with the borate compound Tim-Bor (a commonly used lumber preservative) and absence of a food source (starved termites), using choice and no-choice tests. All wood slices were prepared at the same time using a 10 g liter(-1) solution of each substance and were tested in four different sessions over one year to investigate longevity of the effects. Termites had to tunnel through sand to exploit the food sources, consisting of two wood slices, placed on opposite sides of the experimental enclosure. No-choice tests showed that in the presence of vetiver oil or nootkatone, tunneling activity was always the lowest; wood consumption, termite survival and flagellate numbers and species distribution were significantly different from the control and similar to the results obtained for starved termites and with Tim-Bor-treated wood. Nootkatone negatively affected termites for 12 months and was longer lasting than vetiver oil. In choice tests, when vetiver oil or nootkatone were present, termites exhibited a significant preference for non-treated wood. Our results confirmed both the toxicity and absence of repellency of Tim-Bor. Vetiver oil and especially nootkatone affected Formosan subterranean termites and their protozoa, acting as arrestants, repellents and feeding deterrents, and represent a promising natural alternative for the control of this invasive pest. PMID- 12558099 TI - Synthesis and insecticidal activity of benzoheterocyclic analogues of N'-benzoyl N-(tert-butyl)benzohydrazide: Part 3. Modification of N-tert-butylhydrazine moiety. AB - Nineteen analogues were synthesized by modifying the tert-butylhydrazine moieties of N'-tert-butyl-N'-(3,5-dimethylbenzoyl)-5-methyl-2,3-dihydro-1,4-benzodioxine-6 carbohydrazide and N'-tert-butyl-N'-(3,5-dimethylbenzoyl)-5-methylchromane-6 carbohydrazide (chromafenozide), and the synthesized analogues were evaluated for their insecticidal activity against Spodoptera litura F. While all of the synthesized analogues had insecticidal activity inferior to those of the lead compounds, several of the analogues nonetheless showed high insecticidal activity. Chromafenozide has shown very high selectivity toward lepidopteran species. PMID- 12558101 TI - Variation of pesticide sorption isotherm in soil at the catchment scale. AB - The variation of the sorption isotherm of pesticides has seldom been explored at the catchment scale. Such a study was conducted at the scale of a 187-ha agricultural catchment for three herbicides: atrazine, isoproturon and metamitron. Partition coefficient (Kd) values were measured in batch experiments on 51 topsoil samples, and showed moderate variability at the catchment scale (coefficient of variation CV approximately 30%). Values of Kd ranged from 0.47 to 1.70 litre kg(-1) for atrazine, 0.47 to 1.81 for isoproturon, and 0.55 to 2.21 for metamitron. A clustering method was used to reduce the number of samples on which to measure sorption isotherms to 14. Sorption isotherms agreed with the Freundlich rather than the linear model. Kf parameters had CV values similar to those for Kd, with values ranging from 0.78 to 2.13 mg(1 - Nf) litre(Nf) kg(-1) for atrazine, 0.61 to 1.82 for isoproturon, and 0.69 and 2.58 for metamitron. Nf exponents showed little variation (CV < 5%). Nf values were between 0.86 and 0.98 for atrazine, 0.85 and 0.90 for isoproturon, and 0.82 and 0.87 for metamitron. More than 97% of the Kf catchment-scale variations could be explained by the variations of the soil organic carbon content. PMID- 12558102 TI - Mechanisms of resistance to organophosphorus insecticides in populations of the obliquebanded leafroller Choristoneura rosaceana (Harris) (Lepidoptera: Tortricidae) from southern Ontario. AB - Populations of Choristoneura rosaceana (Harris) from orchards in Ontario were shown to be resistant to azinphos-methyl and to other types of organophosphorus insecticides. Resistance extended to methyl carbamates and to methomyl. The laboratory population used for these assays and selected with azinphosmethyl was also resistant to the pyrethroid, cypermethrin. Resistance was associated with increased esterase activity and was reduced by the addition of the synergist DEF. IEF studies of esterases also indicated increased activity in resistant populations, but did not identify any unique esterases associated with the resistance. Resistance was highly correlated (r = 0.78) with elevated esterases but not with increased glutathione-S transferase activity (r = 0.13). Other mechanisms did not appear to be related. PMID- 12558103 TI - Movement of pendimethalin, ioxynil and soil particles to field drainage tiles. AB - Knowledge of the movement of herbicides and soil particles to sub-surface tile drainage may help to predict chemical leaching to surface waters and deeper groundwater systems. The movement of pendimethalin (2 years), ioxynil (1 year) and soil particles (3 years) to two tile drains was investigated on a sandy loam soil under natural weather conditions. Herbicide and particle concentrations in the drain water showed a very dynamic pattern. The largest herbicide concentrations were detected during the first tile drain flow events after application. Very little herbicide was lost with drain water later than 2 months after application. The turbidity, reflecting concentrations of soil particles, correlated positively and strongly with the pendimethalin concentration and negatively with the rate of drain water discharge, whereas it was uncorrelated with the ioxynil concentration. Peak turbidity values occurred during or shortly after rainfall events, either in break of frost situations, or on unfrozen soil coinciding with the occurrence of peak moisture contents in the topsoil well (3 7%) above field capacity. On average, 0.0013% of the applied pendimethalin and 0.0015% of the applied ioxynil were lost with drain water. The results suggest that preferential flow promotes the movement of all three substances to the tile drains but indicate somewhat different transport mechanisms for the two herbicides. PMID- 12558104 TI - Susceptibility of field populations of adult Bemisia tabaci Gennadius (Homoptera: Aleyrodidae) and Eretmocerus sp (Hymenoptera: Aphelinidae) to cotton insecticides in Burkina Faso (West Africa). AB - Research was conducted in 14 cotton fields (3-10 ha) selected in seven localities (two fields per locality) in Burkina Faso, with the objectives of: (1) estimating Bemisia tabaci Gennadius (Homoptera: Aleyrodidae) population abundance, (2) assessing the levels of parasitism by Encarsia spp and Eretmocerus spp (Hymenoptera: Aphelinidae) and (3) estimating the susceptibilities of the pest and of an Eretmocerus sp to the insecticides currently sprayed on cotton. Yellow sticky cards and a leaf-turning technique were used to estimate adult B. tabaci population densities. Yellow sticky cards were also used to estimate the densities of adult Eretmocerus sp and the susceptibilities of B. tabaci to insecticides. Leaf disk techniques were used to estimate B. tabaci red eye nymph populations and parasitism by Encarsia spp and Eretmoceus spp was evaluated using stereo-microscopy. A leaf cage technique was used to estimate the susceptibilities of Eretmocerus sp to insecticides. A mean of 6.5-27.4 adult B. tabaci were trapped per yellow sticky card and 5.5 to > 34.9 were counted per leaf using the leaf turning technique. There were 0.14-13 Eretmocerus sp trapped per yellow sticky card. The levels of parasitism varied between 36 and 87% by the end of the season and parasitism by Eretmocerus sp predominated in most of the fields. The susceptibilities of B. tabaci and Eretmocerus sp varied from field to field and with the insecticide tested. PMID- 12558105 TI - Diaryloxyacetate herbicides. AB - The discovery and investigation of a novel family of herbicides containing a diaryl acetal are described. The stability of the acetal limited herbicidal efficacy and recognizing that fact led to the design of analogs with commercial levels of post-emergence activity on broadleaf weeds. These compounds inhibited acetolactate synthase and in vitro activity data were used to guide target design. However, no members of this family provided a commercially valuable combination of herbicidal efficacy and crop selectivity. PMID- 12558106 TI - Dissipation of the herbicide dithiopyr in soil and residues in wheat (Triticum aestivum L) grain under Indian tropical conditions. AB - Dissipation of dithiopyr in soil was monitored after application to wheat crop as pre- or post-emergence applications at two rates, viz 100 and 200 g AI ha(-1). The level of dithiopyr in the soil was assessed by gas chromatography, and its disappearence was found to follow a first-order decay curve irrespective of rate or method of application. The half-life in soil ranged between 17.3 and 25.0 days and residues at harvest (150 days after application) ranged between 4.0 and 8.8% of amounts applied. Investigation of microbial degradation of dithiopyr was conducted in minimal salt and Czapek Dox media in which 80% of the compound degraded within 15 days. Residues were not detected in wheat grain at harvest. PMID- 12558107 TI - Acaricidal activity of Cnidium officinale rhizome-derived butylidenephthalide against Tyrophagus putrescentiae (Acari: Acaridae). AB - The acaricidal activity of materials derived from the rhizome of Cnidium officinale Makino against Tyrophagus putrescentiae Schrank adults was examined using direct contact application and fumigation methods, and compared with that of benzyl benzoate and N,N-diethyl-m-toluamide (DEET). The biologically active constituent of the Cnidium rhizome was identified as butylidenephthalide by spectroscopic analyses. On the basis of 24-h LD50 values, acaricidal activity was more pronounced with butylidenephthalide (5.80 microg cm(-2)) than with benzyl benzoate (9.75 microg cm(-2)) and DEET (16.26 microg cm(-2)). In typical poisoning symptoms, butylidenephthalide resulted in lethargy of treated mites, leading to death without knockdown, whereas benzyl benzoate and DEET caused death associated with uncoordinated behaviour. In a fumigation test with T. putrescentiae adults, butylidenephthalide was much more effective in closed containers than in open ones, indicating that the effects of this compound were largely due to action in the vapour phase. Naturally occurring C. officinale rhizome-derived materials merit further study as potential T. putrescentiae control agents or as lead compounds. PMID- 12558108 TI - The Prader-Willi syndrome and the Angelman syndrome. AB - The Prader-Willi syndrome and the Angelman syndrome are characterised by a complex clinical and behavioural phenotype resulting from loss of paternal or maternal expression, respectively, of genes located on the human chromosome 15q11 13. Different molecular mechanisms leading to this imbalance have been identified, including microdeletions, intragenic mutations, uniparental disomy and imprinting centre defects. Low copy repeat gene clusters are known to flank the 15q11-13 microdeletion. They predispose to unequal crossing-over events resulting in the deletion. Involvement of multiple disease genes is strongly suspected and traditional positional cloning techniques as well as animal models are used to identify the involved genes. In this review we include the present state of art and a delineation of future approach to study the candidate genes in these two syndromes. PMID- 12558109 TI - Hypertrichosis cubiti: two new cases and a review of the literature. AB - Two cases of hypertrichosis cubiti in combination with short stature, facial dysmorphias and retarded development are reported with a review of the literature. Hypertrichosis cubiti, the hairy elbows syndrome, consists of a localized form of long vellus hair on the extensor surfaces of the distal third of the upper arm and the proximal third of the forearm bilaterally. It can be associated with short stature and other physical abnormalities. The mode of inheritance has not been established yet; an autosomal recessive as well as an autosomal dominant inheritance trait are postulated. PMID- 12558110 TI - A female with Coffin-Lowry syndrome and "cataplexy". AB - Coffin-Lowry syndrome (CLS) is an X-linked semidominant condition, caused by mutations in the gene encoding the ribosomal protein S6 kinase-2 (RSK-2), a growth factor regulating protein kinase, which is mapped to Xp 22.2. The syndrome is mainly seen in males. It is manifested by moderate to severe mental retardation and characteristic facial, hand and skeletal malformations. We present a female patient with fully manifested CLS, confirmed by molecular analysis, who experienced daily drop episodes, diagnosed as "cataplexy". The episodes were precipitated by emotional or auditory stimuli and were significantly reduced, by selective serotonine re-uptake inhibitors. PMID- 12558111 TI - Partial proximal trisomy 10q syndrome: a new case. AB - We report a case of partial proximal trisomy of the long arm of chromosome 10 confirmed by fluorescence in situ hibridization (FISH) performed with whole chromosome 10 specific painting and specific yac clones. The phenotypic findings, compared to those found in other published cases with the same karyotype, support the recognition of a distinctive partial proximal trisomy 10q syndrome (10q11- >q22). PMID- 12558112 TI - Small terminal 10q26 deletion in a male patient with Noonan-like stigmata: diagnosis by cytogenetic and FISH analysis. AB - A male patient is reported with terminal 10q26 deletion, developmental retardation, special behaviour, and multiple clinical anomalies including hypotonia, short stature of postnatal onset, short webbed neck, craniofacial dysmorphism, pectus excavatum with widely spaced small nipples, cryptorchidism with scrotal hypoplasia, limb and musculoskeletal anomalies. The facial dysmorphism mainly consisted of trigonocephaly, a long, triangular and asymmetrical face, hypertelorism with pseudoepicanthus, broad nasal bridge, high arched palate, retrognathia, low-set dysplastic auricles and, on ophthalmologic examination, strabismus, astigmatism and myopia. Some of these clinical stigmata were suggesting the diagnosis of Noonan syndrome. The extremities showed special features including shortening of proximal limbs, brachydactyly with syndactyly of toes II-III and left fingers III-IV, hypoplastic toenails and joint abnormalities. A diastasis of abdominal muscles was noted and, on X-rays a thoracic scoliosis and bilateral coxa valga were evidenced. Analyses of G- and T banded chromosomes complemented by FISH analyses using different subtelomere probes detected a terminal 10q26 deletion. Subsequent FISH studies using different probes of the 10q26 region were performed in an attempt to closely define the breakpoint and the extent of the deletion and, thereby, to allow karyotype/phenotype comparison between this patient and a previously reported case with an apparently similar 10q26 deletion. PMID- 12558113 TI - Proteus syndrome: a case with clonal chromosome aberration. AB - Proteus syndrome is a disorder characterized by overgrowth of multiple tissues, connective tissue nevi, epidermal nevi and hyperostoses with asymmetric involvement. The clinical expression of the disorder is extremely variable. Molecular pathogenesis of the syndrome is unknown but it is hypothesized that it resulted from a somatic alteration of a gene leading to mosaic effects that would be lethal if the mutation was carried in nonmosaic fashion, and this may explain the variability among patients. We report a new case who presented at birth with asymmetric hypertrophy of the bones and soft tissues of fingers and a tumor of the chest. Cytogenetic analysis of the excised tumor revealed clonal chromosome aberration: mos46, XY, add(9)(p13) [5]/46,XY[30]. During follow up tumors of the rectum and urinary bladder were diagnosed. PMID- 12558114 TI - Partial trisomy 20q in a newborn with dextrocardia. AB - A female newborn is reported with dextrocardia and a partial trisomy 20q, derived from a t(2;20) paternal translocation. The most discriminating findings of the condition include brachycephaly, bulging forehead, deep set eyes, short nose, large ears, dimpled chin, short neck and a heart defect. Previously reported patients with this rare chromosomal anomaly are reviewed. PMID- 12558115 TI - Two brothers with atypical syndactylies, cerebellar atrophy and severe mental retardation. AB - Two brothers with a combination of atypical syndactylies, cerebellar atrophy and severe mental retardation are described. These cases share important features with the group of craniodigital syndromes and could represent new occurrences of Filippi syndrome. Cerebellar atrophy may represent a yet unreported finding in this syndrome. PMID- 12558116 TI - Unilateral absence of the trapezius and pectoralis major muscle: a variant of Poland syndrome. AB - We report a female patient with unilateral absence of the trapezius and pectoralis major muscles without other associated limb abnormalities. We believe that this anomaly belongs to the spectrum of anomalies resulting from disruption of the blood supply in the embryonic subclavian and vertebral arteries, as suggested in Poland syndrome. PMID- 12558117 TI - Tissue specific mosaicism of trisomy 9 in a patient with severe torsion scoliosis. AB - We report on a female patient who had mosaic trisomy 9, presenting with severe scoliosis and mental retardation. Scoliosis is seldom reported in patients with mosaic trisomy 9 syndrome. FISH studies in our proband detected no trisomic cell line in the paravertebral muscle. PMID- 12558118 TI - Sudden death of a girl with Prader-Willi syndrome. AB - We report on the sudden death of a 3.5-year-old girl with Prader-Willi syndrome (PWS) and 15q11-q13 deletion. She suffered from severe chronic breathing disturbances and recurrent bronchitis. During an episode of acute bronchitis she had a cardiac arrest and died two months later of the sequelae. Brain CT imaging three weeks after the arrest showed bilateral symmetrical haemorrhages in the basal ganglia region. The spatial distribution of the haemorrhages can possibly suggest that the basal ganglia in PWS may be especially susceptible to hypoxemia. PMID- 12558119 TI - Dysmorphic face in two siblings with infantile neuroaxonal dystrophy. AB - Infantile neuroaxonal dystrophy (INAD) is an autosomal recessive, neurodegenerative disease with onset in the first or second year of life. It has been reported that INAD shows numerous phenotype characteristics including problems associated with vision, hearing and physical coordination. It has however been very rare to see facial dysmorphism in these children. The study analyzes a girl and boy of a first cousin marriage with infantile neuroaxonal dystrophy affected at birth. At infancy, the children were examined in the Cerrahpasa Medical Faculty Genetic Research Center, Istanbul. They had typical INAD features such as the lack of head control, vision, speech, sitting, and walking which are also seen in children with other congenital abnormalities. These children showed remarkable dysmorphism in the face which included prominent forehead, strabismus, small nose, fish mouth (boy), micrognathia, and large and low-settled ears. The presence of these facial features makes the patients appear unique and diagnosis more accurate. While these features are commonly seen diagnosis may be difficult at its onset. Until now this appearence has not been reported in INAD patients. In conclusion, in the first few months of life without any clinical or neurological signs, the physician should also consider diagnosing the disease of the infant as INAD. PMID- 12558120 TI - "A new association of mental retardation, short stature, unusual face, radio ulnar synostosis and retinal pigment abnormalities": Cohen syndrome with thrombocytopenia. PMID- 12558121 TI - Defective clavicles in Kabuki syndrome. PMID- 12558122 TI - Treatment of no-reflow phenomenon with verapamil after primary stent deployment during myocardial infarction. AB - No-reflow phenomenon is the absence of myocardial perfusion despite adequate dilatation of the infarct related coronary artery during percutaneous coronary intervention. It predicts severe left ventricular dysfunction and poor prognosis in acute myocardial infarction (AMI). The present case is a 54 year old Turkish female who presented with chest pain that had started 2.5 hours earlier. The clinical and laboratory findings were consistent with AMI and the coronary angiogram performed for primary angioplasty revealed a 95% thrombotic occlusion with a TIMI grade I flow in the left anterior descending (LAD) coronary artery. A TIMI grade III flow was achieved with direct stent deployment. However, after the placement of a second stent for severe ostial stenosis more proximally and adjacent to the first one, the antegrade flow became TIMI grade O. As the intracoronary medications did not improve the flow, a mechanical occlusion was considered and a third stent was deployed covering the first two stents. A control angiogram revealed the persistence of TIMI grade O flow. A severe and persistent vasospasm was considered at this point and accordingly, intracoronary verapamil was administered in high concentrations by an infusion catheter to the distal LAD which was followed by the immediate achievement of TIMI grade III flow. Intracoronary administration of high dose verapamil can be performed to prevent vasospasm in resistant no-reflow cases with no evidence of mechanic occlusion. PMID- 12558123 TI - Neointima in coronary stent does not increase during over 1-year in non restenosed lesion at 6 months follow-up: serial volumetric intravascular ultrasound study. AB - The long-term outcomes of coronary artery stenting have been determined by coronary angiography only with has the limitation of determining stent expansion and neointimal proliferation at long-term follow-up. Volumetric intravascular analysis has the potential to evaluate the morphology and distribution of neointima longitudinally after coronary artery stenting. We used three dimensional intravascular ultrasound (3-D IVUS) to evaluate serial changes in stent and neointimal volumes for over 1-year in 9 patients who did not exhibit angiographic restenosis at 6-month follow-up. Volumetric analysis by a validated Netra 3-D IVUS system was performed pre- and post-intervention, at 6-month follow up (FU1), and at over one-year follow-up (FU2). Lumen volume in the stented lesions increased significantly after intervention, and the increase persisted until FU2. There were no significant changes in stent volume between just after stent implantation and at FU2. Neointimal volume within the stents did not change from FUI to FU2 (FU1; 38.4 +/- 9.0 mm3 vs FU2; 33.8 +/- 10.3 mm3). In 33% (3/9) of all lesions, neointimal volume increased between from 6-months to over 1-year after stent implantation. Neointimal distribution after stenting seemed to be almost equal and unrelated to the plaque burden at pre-intervention. Neointimal volume within the stents did not increase and stent volume did not change over the 1st-year in patients who did not exhibit angiographic restenosis at 6-months. PMID- 12558124 TI - Inflammation and immune system response against unstable angina and its relationship with coronary angiographic findings. AB - The aim of this study was to assess the relations between inflammation, immune response, and coronary angiographic findings in patients with unstable angina pectoris (UAP). Recent studies suggest a role for inflammation in the pathophysiology of UAP. Although activation of neutrophils, monocytes and lymphocytes has been shown in UAP, no studies have correlated the activation findings with clinical and angiographic features of patients with UAP. Seventy three patients undergoing coronary angiography were classified according to their ischaemic syndrome, stable angina pectoris (SAP) (n = 25) and UAP (n = 48). Patients with UAP were classified using the Braunwald classification; UAP class I (n = 15), UAP class II (n = 15), and UAP class III (n = 18). Patients with UAP were also classified into a progression to myocardial infarction (MI (+)) group (n = 15) and a non-progression to myocardial infarction (MI(-)) group (n = 33). Venous blood samples were taken from all patients. Cell surface receptors (CD4, CD8, CD3, CD14, CD45, CD56+16, and HLA-DR) were detected by flow cytometry using monoclonal antibodies tagged with fluorescent markers and serum levels of C reactive protein (CRP) were measured. The serum levels of CRP and the percentages of HLA-DR, CD14, and CD16+56 were higher in UAP than SAP. The serum levels of CRP and percentages of HLA-DR, CD14, and CD16+56 were higher in UAP class II than UAP class I. The serum levels of CRP and percentages of HLA-DR, CD14, and CD16+56 were higher in UAP class III than UAP class II and UAP class I. The serum levels of CRP and percentages of CD16+56 were higher in the MI(+) group than the MI(-) group. The CRP levels in serum and the percentages of cell surface antigens had no correlation with extent of coronary artery disease (no differences among one, two or three vessels) but Type C lesion had significantly higher percentages of HLA-DR, CD14, CD16+56 and the serum levels of CRP than Type A and Type B lesions. This investigation shows that inflammatory and immunologial components may be detectable in UAP and were correlated with the clinical severity, progression to myocardial infarction, and lesion morphology, but were not correlated with the extent of coronary artery disease. PMID- 12558125 TI - C-reactive protein and coronary artery disease. AB - Evidence suggests that inflammation plays a key role in the pathogenesis of atherosclerosis. The chronic inflammatory process can develop to an acute clinical event by the induction of plaque rupture and therefore cause acute coronary syndromes. The aim of this study was to determine the serum levels of the circulating acute-phase reactant C-reactive protein (CRP), which is a sensitive indicator of inflammation, in patients with chronic stable coronary artery disease (CAD) and acute coronary syndromes (ACS). We studied 56 subjects: 1) 25 consecutive patients (18 men, 7 women; mean age, 68.5 +/- 14.3 years, range, 40-86) with unstable angina (UA) or acute myocardial infarction (AMI); 2) 31 consecutive patients (25 men, 6 women; mean age 64 +/- 12.7; range, 47-83, years) with signs and symptoms of clinically stable CAD. High-sensitivity-C reactive protein (hs-CRP) levels were determined with a commercially available enzyme-linked immunoassay method. In patients with unstable angina and AMI before reperfusion therapy, CRP levels were not significantly different to those in patients with stable CAD (5.96 +/- 2.26 versus 4.35 +/- 2.6 mg/L; P = 0.12), but tended to be higher in patients with unstable angina and AMI. Baseline CRP levels in the subgroup of patients with AMI (6.49 +/- 2.28 mg/L) were significantly higher than levels in patients with stable CAD (4.35 +/- 2.6 mg/L; P = 0.02). CRP levels in patients with unstable angina and AMI were measured four times during a 72-hour period (0, 12, 24, and 72 hours). The lowest value was observed at baseline and differed significantly from values measured at any other time of the observation period (P < 0.001; 5.96 +/- 2.26; 9.5 +/- 9.04, 18.25 +/- 11.02; 20.25 +/- 10.61). CRP levels after 12, 24, and 72 hours were also significantly different to the initial values for patients with stable CAD (P < 0.01). There was no correlation between CRP and creatine kinase (CK), CK-MB isoenzyme, or troponin I positivity as markers for the extent of the myocardial injury during the observation period. Baseline levels of serum CRP tended to be higher in patients with unstable angina or AMI but were not significantly different from levels in patients with chronic stable CAD. In the subgroup of patients with AMI, baseline CRP levels were significantly higher than the levels in patients with stable CAD. CRP as a marker of inflammation is significantly increased in patients with AMI and unstable angina shortly after the onset of symptoms (after a period of 12 hours), supporting the hypothesis of an activation of inflammatory mechanisms in patients with an acute coronary syndrome or AMI. PMID- 12558126 TI - Comparison of cardiovascular responses to isometric (static) and isotonic (dynamic) exercise tests in chronic atrial fibrillation. AB - The aim of the present study was to evaluate the tolerance to various exercises by determining the cardiovascular response to static and dynamic exercises in patients with nonvalvular atrial fibrillation. Fifty patients (mean age: 63.6 +/- 10.3 years; male: 25, female: 25) with chronic (more than one year) nonvalvular atrial fibrillation were included in the study. All patients underwent exercise tests, adjusted appropriately according to their symptoms, as dynamic exercise on a Marquette Case 15 device according to a modified Bruce protocol. Heart rate, and systolic and diastolic arterial pressures were measured at rest and at all stages of the exercise; and the heart rate-pressure products were evaluated. A handgrip test was also conducted as static exercise. The measurements were made before, at the 1st, 2nd and 3rd minutes, and in the recovery periods of the exercise. The percent values of the changes of the 1st, 2nd and 3rd minute measurements in relation to the initial values for both exercises were compared. In addition, the maximal responses to the exercise tests and the post exercise values were also compared. For statistical evaluations, the paired Student-t test was used. Heart rate and pressure-heart rate product values obtained at 1, 2, and 3 minutes during the treadmill exercise test were significantly high compared to the handgrip values (P < 0.0001). The arterial systolic and diastolic pressure values in the 1st minute were also significantly higher during the handgrip test (P = 0.0100 and P = 0.0320, respectively). The values of diastolic arterial pressure at the 2nd minute during the handgrip test, and systolic arterial pressure at the 3rd minute during the treadmill test were found to be statistically significant (P = 0.0240, P = 0.0340, respectively). The mean exercise time and MET value during the treadmill exercise test were 7.18 +/- 2.65 minutes and 5.32 +/- 1.38 mL.kg(-1) x dk(-1). respectively. During the recovery period, the 5th minute, heart rate and pressure-heart rate product values were significantly high after the treadmill test (P < 0.0001). In this study, we revealed that the heart rate response to static exercise was lower and the patients tolerated the static exercise better. Therefore, we decided that the short duration of static exercise is not harmful for the noncomplicated chronic atrial fibrillation cases. PMID- 12558127 TI - Long-term follow-up of pacemakers with an Autocapture pacing system. AB - The aim of this study was to evaluate the safety and performance of the Autocapture pacing system during a 5-year follow-up period. The study was conducted retrospectively between May 1996 and May 2001. Sixty consecutive patients who had undergone VVI pacemaker implantation using an Autocapture program with leads 1402T (n: 31) and 1452T (n: 29) were included in the study. Intraoperative measurements including a ventricular stimulation threshold test, sensing of intrinsic R wave (mV), and lead impedance (W) were done by a standard pacing system analyzer. Evoked responses (ER, mV) and polarization signals (PS, mV) were measured after the pocket was closed. Pacing thresholds by Autocapture (AC thrd, V) and Vario (Vario thrd, V), battery current (mA), and battery impedance (kW) were also repeated during predischarge and 1, 6, 12, 18, 24, 30, 40, 50, and 60 months after discharge. According to the ER and PS values an Autocapture algorithm could be activated in 49 patients (88%). The Autocapture algorithm remained active during the follow-up in all of these patients. In patients with inappropriate ER and PS values (11 patients, 12%), pacemakers were programmed to a VVIR pacing mode and Autocapture algorithm was inactivated. ER and PS values did not reach appropriate values to activate the Autocapture algorithm in any of these patients in consecutive follow-ups. Twenty-four-hour Holter monitoring could be conducted in 32 patients (53%). In all recordings, when the loss of capture occurred, it was confirmed that back-up pacing continued. When the first measurements recorded during implantation were compared to approximately the 5th year measurements; ER (9.2 mV vs 9.6 mV), PS signal (1.13 +/- 0.30 mV vs 1.15 +/- 0.72 mV), AC thrd (0.4 V vs 1.2 V), Vario thrd (0.7 V vs 1.3 V), and lead impedance (502 ohm vs 620 ohm) were not changed significantly. Battery impedance increased 1 kOhm between 30-40 months of the implantation. Seven deaths occurred during follow-up. Three patients had fatal myocardial infarction, one died due to a non-cardiac event, and the remaining three died due to progressive heart failure. CONCLUSION: ER, R wave amplitude, and PS, which are the main parameters for the continuation of Autocapture function, did not change significantly during long-term follow-up. High output back up pacing provided additional safety for sudden rises in threshold. The Autocapture pacing algorithm was found to be effective and reliable during long term follow-up. PMID- 12558128 TI - Clinical characteristics of induced nonclinical ventricular tachycardia in nonischemic cardiomyopathy. AB - The clinical significance of induced nonclinical ventricular tachycardia (NCVT) in nonischemic dilated cardiomyopathy (DCM) remains controversial. Twenty-eight patients with sustained VT or ventricular fibrillation related to DCM underwent programmed ventricular stimulation (PVS) to induce VT. However, VT was not induced in four patients. Based on the morphology of induced ventricular arrhythmia, we classified the remaining 24 patients into NCVT (n = l1 ) and clinical VT (CVT) groups (n = 13), then evaluated the prognosis for a mean follow up period of 22 months. The cycle length of induced NCVT was significantly shorter than that of induced CVT (277 +/- 38 ms vs 325 +/- 63 ms, P < 0.05). Appropriate antiarrhythmic agents were selected by serial PVS in 36% of the NCVT group and in 38% of the CVT group (4/11 vs 5/13). Among patients who had been treated by PVS guided drug therapy, arrhythmic events were observed in 75% of the NCVT group and 80% of the CVT group (3/4 vs 4/5). The total incidence of sudden death in the NCVT group was higher than that in the CVT group (5/11: 45% vs 4/13: 31%). In conclusion, induced NCVT and CVT are refractory to pharmacological therapy and both have an important characteristic value in DCM. PMID- 12558129 TI - The polymorphisms of codon 727 and 52 of thyroid-stimulating hormone receptor gene are not associated with mitral valve prolapse syndrome in Taiwan Chinese. AB - A germline mutation of the thyroid-stimulating hormone receptor (TSHR) gene has been reported to be associated with thyrotoxicosis and mitral valve prolapse syndrome (MVPS) in a Chinese family. The role of TSHR genetic variants in MVPS has not been well studied. This study investigated the possible relationship between the polymorphisms of codon 727 and 52 of the TSHR gene and MVPS among the Chinese population in Taiwan. We studied 100 patients with MVPS diagnosed by echocardiography and 100 ageand sex-matched normal control subjects. The polymorphisms of codon 727 and 52 of the TSHR gene were identified by polymerase chain reaction-based restriction analysis. There was no significant difference in either the genotype distribution or allelic frequencies between MVPS cases and controls for either TSHR gene D727E polymorphism ( = 0.51 and 0.45, respectively ) or P52T polymorphism (P = 0.60 and 0.31, respectively). The MVPS patients were divided into 2 subgroups: those with Graves' disease, and those without the thyroid disorder, and there were no statistical differences from the controls for both the TSHR gene D727E and P52T polymorphisms. Further categorization of the MVPS patients into mild and severe subgroups also revealed no statistical difference from controls for either the TSHR gene D727E or P52T polymorphisms. These findings suggest that the codon 727 and 52 polymorphisms of the TSHR gene are not the suitable genetic markers of MVPS in Taiwan Chinese. PMID- 12558130 TI - Effects of cilazapril on endothelial function and pulmonary hypertension in patients with congestive heart failure. AB - Flow-mediated vasodilation (FMV), brachial artery flow (BAF), and brachial artery diameter were evaluated in 30 patients with congestive heart failure before and after cilazapril treatment. While mean pulmonary artery pressure and pulmonary capillary wedge pressure decreased significantly, flow-mediated vasodilation and left ventricular ejection fraction increased significantly following cilazapril administration (P < 0.001). Brachial artery diameter and brachial artery flow did not change following the treatment period (P > 0.05). In conclusion, short-term cilazapril administration improved endothelial function and pulmonary pressure in patients with congestive heart failure. PMID- 12558131 TI - Effects of the angiotensin-converting enzyme inhibitor enalapril on sympathetic neuronal function and beta-adrenergic desensitization in heart failure after myocardial infarction in rats. AB - One of the beneficial effects of angiotensin-converting enzyme (ACE) inhibitors in the treatment of heart failure may derive from sympathoinhibition and the prevention of beta-adrenergic desensitization. However, the roles of these properties in the overall effects of ACE inhibitor are not clear. We studied the effects of chronic enalapril treatment (20 mg/L in drinking water for 12 weeks) on left ventricular (LV) function, cardiac norepinephrine (NE), sympathetic neuronal function assessed by 131I-metaiodobenzylguanidine (MIBG), beta receptors, and isometric contraction of papillary muscle in rats with myocardial infarction (MI) induced by coronary artery ligation. Decreased LV function in the MI rats was associated with reduced cardiac NE content and MIBG uptake, and severely blunted responses of non-infarcted papillary muscle to isoproterenol, forskolin, and calcium. Enalapril attenuated LV remodeling in association with a reduction of the ventricular loading condition and restored baseline developed tension of non-infarcted papillary muscle to the level of sham-operated rats. However, enalapril did not improve cardiac NE content, MIBG uptake, or inotropic responsiveness to beta-agonists. These results suggest that the major effect of the ACE inhibitor enalapril in the treatment of heart failure is not due to sympathoinhibition or restoration of beta-adrenergic pathway in this model of heart failure. PMID- 12558132 TI - Volume overload influence on hypertrophied myocardium function. AB - The aim of this study was to demonstrate that hypertrophied cardiac muscle is more sensitive to volume-overload than normal cardiac muscle. We assessed the mechanical function of isolated left ventricular papillary muscle from male spontaneously hypertensive rats (SHR) and age-matched normotensive Wistar-Kyoto rats (WKY) submitted to volume overload caused by aortocaval fistula (ACF) for 30 days. Muscles were perfused with Krebs-Henseleit solution at 28 degrees C and studied isometrically at a stimulation rate of 0.2 Hz. The ACF increased the right and left ventricular weight-to-body weight ratio in WKY rats; it also promoted right ventricular hypertrophy and further increased the basal hypertrophy in the left ventricle from SHR. The arterial systolic pressure was greater in SHR than in WKY rats, and decreased with ACF in both groups. Developed tension (DT) and maximum rate of DT (+dT/dt) were greater in the SHR-control than in the WKY-control (P < 0.05); the time from peak tension to 50% relaxation (RT 1/2) was similar in these animals. ACF did not change any parameters in the SHR group and increased the resting tension in the WKY group. However, the significant difference observed between myocardial contraction performance in WKY controls and SHR-controls disappeared when the SHR-ACF and WKY-controls were compared. Furthermore, RT 1/2 increased significantly in the SHR-ACF in relation to the WKY-controls. In conclusion, the data lead us to infer that volume overload for 30 days promotes more mechanical functional changes in hypertrophied muscle than in normal cardiac muscle. PMID- 12558133 TI - A case of atrioventricular complete block due to Behcet's disease. AB - The cardiovascular system is involved in 5% of the cases of Behcet's disease. Thrombophlebitis. aneurysm in arteries, pericarditis, myocarditis, valvular disease, ventricular arrhythmia, and conduction system disorders may occur. A case of Behcet's disease with complete proximal atrioventricular block is presented. PMID- 12558134 TI - Platelet transfusion and bleeding after cardiopulmonary bypass. PMID- 12558135 TI - Association of a TNFbeta gene polymorphism with complications after major abdominal operations. AB - In injured patients, it has been shown that a polymorphism of the tumor necrosis factor-beta (TNFbeta) gene is related to the development of sepsis. We investigated the relation of TNFbeta gene polymorphism with the development of severe complications after elective major abdominal operations, and with production of TNFalpha perioperatively. In the present investigation, the Ncol polymorphism was studied in genomic DNA isolated from the blood of 172 patients. Preoperatively and postoperatively, lipopolysaccharide (LPS)-stimulated production of TNFalpha in the patients' whole blood was tested in vitro. Genotypes and TNFalpha production were related to the occurrence of severe complications. Postoperatively, 15% (n = 26) of the patients developed severe complications. The overall mortality was 2% (n = 3). The homozygous TNFB2 genotype was found in 54% of the patients, the homozygous TNFB1 genotype was found in 14% of the patients, and the heterozygous genotype was found in 32% of the patients. In patients with complications, the B2B2 genotype was much more frequent (21/26, 81%) than in those without complications (72/146, 49%; P < 0.003). The development of complications was associated with a lower capacity to produce TNFalpha 3 and 7 days after the operation. In patients without complications, the TNFbeta polymorphism was not related to different levels of TNFalpha production. These data indicate an association between TNFbeta polymorphism and postoperative complications and they suggest the B2/B2 genotype as a high risk factor for the development of sepsis after elective operative trauma. PMID- 12558136 TI - Expression patterns of the lipopolysaccharide receptor CD14, and the FCgamma receptors CD16 and CD64 on polymorphonuclear neutrophils: data from patients with severe bacterial infections and lipopolysaccharide-exposed cells. AB - In polymorphonuclear neutrophils (PMN) CD14, one of the receptors for lipopolysaccharides (LPS) is stored intracellularly as a preformed protein, with only few receptors expressed on the surface. We now report that in patients with severe bacterial infections, CD14 expression is profoundly upregulated, as is CD64 (FcgammaRI), the high-affinity receptor for IgG, whereas CD16 (FcgammaRIII) was partly lost from the surface. To further analyze regulation of these receptors, PMN of healthy donors were exposed to low doses of LPS. By brief exposure (10-120 min) to LPS, CD14 was transferred to the surface in a cytochalasin B-sensitive manner, as were CD16 and CD64. Prolonged culture (up to 48 h) resulted in a further upregulation of CD14, sustained expression of CD64, and profound decline of CD16, yielding a similar pattern of receptor expression as seen in the patients. Subsequent studies revealed that LPS induced de novo synthesis of CD14: the increase of surface expression could be inhibited by cycloheximide and by interfering with a known LPS-induced signaling event, the translocation of NFkappaB. Moreover, an up to 10-fold increase of specific mRNA was seen, as was incorporation into CD14 of 35S-methionine. The de novo synthesis prolonged expression of CD14, whereas the CD16 expression declined, generating a PMN phenotype characteristic for severe infection and indicative of escape from apoptosis of a PMN subpopulation. PMID- 12558137 TI - Baseline cortisol levels, cortisol response to corticotropin, and prognosis in late septic shock. AB - The prognostic value of basal and corticotropin-stimulated cortisol concentration in patients with sepsis remains a controversial issue. In a retrospective cohort study, 82 consecutive patients with septic shock underwent a short corticotropin test performed more than 24 h after the onset of vasopressor therapy. Forty-one (50%) patients died within 28 days after the onset of septic shock. The mean (SD) basal cortisol level was 22.7 (10.6) microg/dL. With threshold values of 7 and 9 microg/dL maximal increases in cortisol level, 28 (34%) and 31 (38%) patients were, respectively, classified as nonresponders to the short corticotropin test. On multivariate analysis, a cortisol level >20 microg/dL (P = 0.0002), a maximal response to corticotropin <9 microg/dL (P = 0.044), abnormal lactate values (P = 0.0098), and positive blood cultures (P = 0.004) were independent predictors of 28-day mortality. In conclusion, high basal cortisol and low increase on corticotropin stimulation are predictors of a poor outcome in late septic shock. The underlying mechanisms of these prognostic patterns remain to be elucidated. PMID- 12558138 TI - Transforming growth factor-beta1 and major trauma: time-dependent association with hepatic and renal insufficiency. AB - Dysbalance in the immune system is perceived as a major factor for adverse outcome after trauma. Transforming growth factor-beta1 (TGF-beta1) is a multifunctional cytokine that regulates proliferation, differentiation of cells, wound healing, and angiogenesis. The influence of TGF-beta1 on trauma patients outcome is still unclear. Injury patterns and clinical outcome parameters of 99 consecutive patients with life-threatening injury and an injury severity score (ISS) > 15 were assessed in a prospective, single-center study at a Level I trauma center. Levels of TGF-beta1 in plasma were measured over a 5-day period by an enzyme-linked immunoabsorbant assay (ELISA). TGF-beta1 plasma levels rise shortly after trauma and gradually drop as the 5th day approaches. Mean and maximal TGF-beta1 plasma levels were significantly higher in patients who developed sepsis and were significantly lower in patients with renal or hepatic failure. Receiver operating characteristics-curve analysis of liver failure shows an area under the curve (AUC) of 0.68 (95%: 0.55-0.81, P = 0.02) and of an AUC of 0.63 (95%: 0.52-0.75, P = 0.03) for renal failure for maximal TGF-beta1 plasma (initial until day 2) levels if lower values represent a more positive test. The data indicate that the increase and decrease of TGF-beta1 plasma levels may contribute to clinical outcome after severe injury. Lower TGF-beta1 levels are associated with liver and renal insufficiency. Higher TGF-beta1 levels 6 h after ICU admission increase the risk of sepsis. TGF-beta1 seems to be an early onset reactant and not a second-line responsive cytokine. PMID- 12558139 TI - Monocyte CD40 expression in severe sepsis. AB - D40 is a cell surface protein belonging to the tumor necrosis factor (TNF) receptor family. Ligation of monocyte CD40 by the T cell-derived CD40 ligand can trigger the production of various mediators, the transcription and activation of enzymes, and the upregulation of costimulatory molecules involved in the pathogenesis of sepsis. To test the hypothesis that CD40 is expressed on the surface of monocytes during sepsis, we measured CD40 expression by flow cytometry on freshly sampled monocytes from 40 patients with severe sepsis, including 15 patients with bacteremia, and from eight healthy volunteers. Plasma concentrations of interleukin (IL) 6, IL-10, and IL-13 were also measured. We detected CD40 only on monocytes from patients with sepsis (mean 6.5 +/- 0.4 median channel fluorescence). There was an inverse correlation between peak CD40 expression and survival (P = 0.05), particularly in the patients with bacteremia (P = 0.019). In the bacteremic group, there was an inverse correlation between CD40 expression and bilirubin levels (r2 = 0.52, P = 0.004) and plasma IL-6 concentrations (r2 = 0.30, P = 0.04). Our results showed that upregulation of CD40 expression on peripheral blood monocytes is a protective phenomenon during severe sepsis. Monocyte deactivation reflected by low CD40 expression may represent impairment of immune function associated with severity of illness and poor outcome. Further studies on monocyte phenotype and function may help to assess the immune status of patients with sepsis and perhaps be useful to guide immunomodulatory strategy in the future. PMID- 12558140 TI - Age-related gender differential in outcome after blunt or penetrating trauma. AB - Although laboratory studies indicate that female rodents better tolerate the deleterious consequences of trauma and have higher survival rates than male rodents, it remains unclear whether a similar gender dimorphic pattern is evident in humans. In view of this, the association between gender and mortality in trauma patients admitted to a University Level I Trauma Center was assessed. All adult patients admitted to the University of Alabama at Birmingham Trauma Center with blunt or penetrating injury between July 1996 and March 2001 were selected for analysis. Patients were categorized by mechanism (blunt or penetrating), and odds ratios (ORs) were used to compare the risk of death among males compared with females. The ORs were stratified according to age and were adjusted for demographic, medical, and injury characteristics. Male blunt trauma patients <50 years old had a 2.5 times (95% CI 1.3-4.9) higher risk of death than females; however, for those > or = 50 years old, a smaller, nonstatistically significant difference was apparent (OR 1.4, 95% CI 0.8-2.3). Conversely, for penetrating trauma, males <50 years old exhibited an increased yet nonsignificant risk of death (OR 1.8, 95% CI 0.6-5.4), whereas those > or = 50 years old had a survival advantage (OR 0.1, 95% CI 0.02-0.5). Laboratory studies have demonstrated that estrogens are salutary and androgens are detrimental for survival following trauma-hemorrhage. The results of this study suggest that the physiologic pattern of premenopausal adult female sex hormones may provide a survival advantage in blunt trauma patients; however, the converse pattern prevails for the penetrating trauma patients. PMID- 12558141 TI - Caspase inhibition does not protect against liver damage in hemorrhagic shock. AB - This study was aimed to determine whether administration of an inhibitor of caspase-3 protects hepatocellular function in rats with hemorrhagic shock and whether caspases are important pharmacological targets in attenuating liver injury induced by hemorrhagic shock and resuscitation. Male adult rats were subjected to hemorrhagic shock by bleeding to a mean arterial blood pressure of 35-40 mmHg for 1 h and were then resuscitation with 60% shed blood and lactated Ringers solution. A subgroup of animals was injected i.v. with 2 mg/kg caspase inhibitor, Z-DEVD-FMK, prior to blood withdrawal. Fas ligand expression was markedly elevated and caspase-3 activity increased by 3-fold in hemorrhagic untreated rats. The increase in caspase-3 activity was prevented by administration of Z-DEVD-FMK prior to shock and resuscitation. Poly (adenosine diphosphate ribose) polymerase proteolysis was reduced in rats treated with the caspase-3 inhibitor compared with hemorrhagic untreated animals. Plasma aspartate aminotransferase and alanine aminotransferase values showed a significant increase at 6 h of shock in untreated animals (+360% and +515% as compared with sham-operated animals, respectively). Administration of the caspase-3 inhibitor did not prevent the increase in plasma transaminases. The cytosolic concentration of thiobarbituric acid-reactive substances (TBARS) and the oxidized:reduced glutathione ratio increased in the animals with hemorrhagic shock (+94% and +170%, respectively). These parameters were not significantly modified by pretreatment with Z-DEVD-FMK. It appears that caspase inhibition does not attenuate hepatocellular depression and liver injury induced by hemorrhagic shock and resuscitation. PMID- 12558142 TI - Hemodynamic effects of early versus late glucocorticosteroid administration in experimental septic shock. AB - Recent findings in human septic shock suggest that glucocorticosteroids can limit and even reverse hemodynamic disturbances and dependence on catecholamines. In a rodent model of hypotensive and hypokinetic septic shock, we investigated the effects of early or late dexamethasone administration on hemodynamics, response to catecholamines, and cardiac beta-adrenergic signalling. As compared with sham operated rats, the untreated septic rats displayed significant arterial hypotension and reduced aortic blood flow. However, in vivo pressor response to epinephrine and phenylephrine was not different among sham and septic animals. Conversely, the chronotropic response to isoproterenol was significantly attenuated in septic animals. Steroid-treated septic animals displayed complete reversal of hypotension, improvement in aortic blood flow, and reduced plasma lactate and nitrite/nitrate concentrations as compared with untreated septic animals. The number of myocardial beta-adrenergic receptors and in vivo isoproterenol-stimulated myocardial cAMP content were similar in sham and septic animals. Glucocorticosteroids, although not changing these patterns, significantly decreased the receptors affinity when administered late, but not early. In this model of septic shock, hemodynamic abnormalities may not be related to adrenergic receptor desensitization. That steroids can improve them suggests that they could act mainly distal to adrenergic receptors, for instance, on myocardial and vascular smooth fiber contraction properties through mechanisms probably including inducible nitric oxide synthase inhibition. PMID- 12558143 TI - Transcription factors C/EBP-alpha and HNF-1alpha are associated with decreased expression of liver-specific genes in sepsis. AB - Previous studies have demonstrated sepsis-specific changes in the transcription of key hepatic genes. However, the role of hepatic transcription factors in sepsis-associated organ dysfunction has not been well established. We hypothesize that the binding activities of C/EBPalpha and beta, HNF-1alpha, and HNF-3 transiently decrease during mild sepsis but persistently decrease after fulminant sepsis, and that the decrease in this binding activity correlates in time and severity with previously described decreases in the transcription of key hepatic genes. Male C57/BL6 mice had nonlethal sepsis induced by cecal ligation and single puncture (CLP) and fulminant sepsis via cecal ligation and double puncture (2CLP). Sham-operated and unoperated animals served as controls. Transcription factor binding activity was assessed with electrophoretic mobility shift assays. C/EBP-a and HNF-1alpha binding activity decreased transiently after CLP and persistently after 2CLP. Binding activity of both C/EBP-beta and HNF-3 were unchanged. The decrease in C/EBP-a and HNF-1alpha binding activities correlated in time and magnitude with the decreased hepatic gene transcription previously observed in sepsis. Furthermore, the loss of activity after 2CLP correlated in time with outcome. Sepsis decreases DNA binding activities of C/EBPalpha and HNF 1alpha, two key hepatocyte transcription factors, in a time course consistent with down-regulation of their target hepatic genes. Therefore, alterations in transcription factor binding are likely important in the transcriptional modulation that is characteristic of hepatic dysfunction in sepsis. PMID- 12558145 TI - Effects of fluid resuscitation on mesenteric microvascular blood flow and lymphatic activity after severe hemorrhagic shock in rats. AB - We investigated the acute microcirculatory effects, including mesenteric lymphatic pumping, of volume replacement with different iso- or hypertonic/oncotic solutions after severe hemorrhage (mean arterial pressure [MAP] approximately 35 mmHg during 30 min) in halothane-anesthetized Wistar rats. Resuscitation was achieved 30 min after induction of shock with one of the following solutions: autologous blood (BL); 0.9% NaCl (IS), 7.5% NaCl (HS); 5% bovine serum albumin (BSA); 0.9% NaCl-6% hydroxyethyl starch (HES), or 7.5% NaCl HES (HES 7.5). MAP was partially and transiently restored by infusion of IS or HS, whereas in the groups treated with BL, HES, HES 7.5, or BSA, there was a complete restoration of blood pressure in the 30-min period after infusion. Microvascular blood flow (MBF), measured by laser Doppler flowmetry, was reduced by 59% +/- 7% 10 min after bleeding. MBF was also transiently restored after infusion of IS, HS, BL, BSA, or HES. HES 7.5 was the only solution able to induce immediate and sustained (60 min) restoration of preshock levels of MBF. Volume replacement with IS or HES 7.5 resulted respectively in long-lasting or transient lymphatic pumping overload. On the other hand, resuscitation with all other solutions, except BSA, did not restore lymphatic activity to preshock levels. We also observed a significant reduction of the diameter of mesenteric terminal arterioles (15-30 microm) after bleeding, which was reversed temporarily in IS, BL, and HES groups, whereas resuscitation with HES 7.5 solution was able to maintain arterioles dilated until the end of the experimental period. Therefore, it is concluded that the association of hyperoncotic and hyperosmotic solutions, represented here by HES 7.5, induces positive effects with respect to the macro- and microhemodynamics accompanied by restoration and maintenance of the interstitial drainage system, being indicated for maintenance of postresuscitation cardiovascular and microvascular function. PMID- 12558144 TI - Glutamine improves impaired cellular exudation and polymorphonuclear neutrophil phagocytosis induced by total parenteral nutrition after glycogen-induced murine peritonitis. AB - Clinical and laboratory evidence shows that enteral feeding significantly reduces pneumonia and intra-abdominal abscess formation after celiotomy for severe trauma. Supplementation of total parenteral nutrition (TPN) with glutamine (GLN) supports impaired immunity induced by TPN in several animal and human studies. This work investigates the peritoneal cellular response and polymorphonuclear neutrophil (PMN) bactericidal function after mouse chemical peritonitis after TPN with and without GLN. Thirty-three mice received chow, TPN, or 2% GLN supplemented TPN (GLN-TPN) for 5 days. All mice then received 2 mL of a 1% glycogen solution intraperitoneally to induce cell exudation, and peritoneal exudative cells (PECs) were recovered 4 h later. Total and differential PEC numbers, as well as PMN phagocytosis, reactive oxygen intermediate production (ROI), CD11b (integrin aM chain) expression, and CD16/32 (Fcgamma II/III receptor) expression were measured. PMN, macrophage, and lymphocyte cell numbers were significantly lower with TPN than with chow or GLN-TPN groups, with no differences between chow and GLN-TPN. TPN significantly lowered peritoneal PMN phagocytosis compared with chow (P < 0.05) and approached significance with GLN TPN (P = 0.06). There were no significant differences in ROI production or CD11b and CD16/32 expression on peritoneal PMN. GLN supplementation improved the reduction in cell exudation and PMN phagocytosis induced by TPN after chemical peritonitis. PMID- 12558146 TI - A critical role of leukotriene B4 in neutrophil migration to infectious focus in cecal ligaton and puncture sepsis. AB - Neutrophil migration to an infectious focus is essential for control and resolution of infection. Early studies demonstrated that the failure of such migration is observed in lethal sepsis induced by cecal ligation and puncture (L CLP), whereas intense neutrophil migration is seen in sublethal CLP (SL-CLP). In this study, we found that inhibition of synthesis of prostaglandins or leukotriene B4 (LTB4) did not modify the failure of neutrophil migration or the survival rate of L-CLP mice. In addition, pretreatment of L-CLP mice with a platelet activating factor (PAF) receptor antagonist (UK74505), despite not interfering with the failure process, significantly increased (33%) the survival rate of the animals. Inhibitors of prostaglandin synthesis (indomethacin and meloxican) and UK74505 did not modify the neutrophil migration observed in SL CLP. On the other hand, the blockade of LTB4 synthesis (MK886, a 5-lipoxygenase activating protein inhibitor) or of its receptors (CP-105,696) resulted in reduced neutrophil migration to the peritoneal cavity in SL-CLP mice (62% and 60%, respectively), a consequent increase in the number of bacteria in the inflammatory focus, and a reduced survival rate of the animals (43% and 38%, respectively). Both SL-CLP and L-CLP animals presented significant levels of LTB4 in the peritoneal exudate (3- and 8-fold higher than sham group, respectively) and these were reduced by the pretreatment of mice with LTB4 inhibitors. In conclusion, our results suggest that LTB4, but not prostaglandins or PAF, is an important chemoattractant involved in neutrophil recruitment to infection sites in SL-CLP, a crucial event in confining the invading pathogens to a restricted area. However, in circumstances in which the infection turns to a lethal sepsis, LTB4 is not involved in the observed failure of neutrophil migration to the infectious focus. PMID- 12558147 TI - Endotoxemia inhibits intestinal adaptation in a rat model of short bowel syndrome. AB - Sepsis is commonly associated with or complicates short bowel syndrome (SBS). The purpose of the present study was to investigate the effects of endotoxemia on intestinal adaptation in a rat model of SBS. Male Sprague-Dawley rats were divided into three experimental groups: Sham rats underwent bowel transection and re-anastomosis, SBS rats underwent 75% small bowel resection, and SBS-LPS rats underwent bowel resection and were given lipopolysaccharide. Bowel weight, organ weights, and parameters of intestinal adaptation (bowel and mucosal weights, mucosal DNA and protein, villus height, and crypt depth) were determined on day 15 following operation. The results of this study demonstrate that SBS rats showed a significant increase (vs. Sham) in jejunal and ileal bowel and mucosal weight, mucosal DNA and protein, villus height, and crypt depth. SBS-LPS animals demonstrated lower (vs. SBS rats) final body weight (215 +/- 7 vs. 287 +/- 10 g, P < 0.05), overall weight in duodenum (98+/- 2 vs. 119 +/-5 mg/cm, P < 0.05) and jejunum (144 +/- 9 vs. 189 +/- 16 mg/cm, P < 0.05), mucosal weight in jejunum (54 +/- 5 vs. 69 +/- 5 mg/cm, P < 0.05) and ileum (31 +/- 2 vs. 37 +/- 3 mg/cm, P < 0.05), mucosal DNA in jejunum (89 +/- 11 vs. 120 +/- 11 microg/cm, P < 0.05) and ileum (46 +/- 6 vs. 61 +/- 4 microg/cm, P < 0.05), jejunal crypt depth (152 +/- 19 vs. 189 +/- 12 microm, P < 0.05), and ileal villus height (405 +/- 63 vs. 515 +/- 30 pm, P < 0.05). In addition, the SBS group had no late (second week) mortality, whereas the SBS-LPS group had an 17% late mortality rate. In conclusion, in a rat model of SBS-LPS, endotoxemia appears to inhibit structural intestinal adaptation and increase mortality. PMID- 12558149 TI - Contribution of myoglobin-induced increases in vascular resistance to shock decompensation in experimental Crush-syndrome in anesthetized rats. AB - Myoglobin is known to become nephrotoxic when released in greater amounts from skeletal muscle into the general circulation during shock. The present study deals with the question as to whether a myoglobin-induced increase in vascular tone additionally contributes to the detrimental role of this protein in hypovolemic shock. Anesthetized rats were subjected to 250 mg kg x h(-1) myoglobin infused i.v. during hemorrhagic hypotension of 50 mmHg. Shock survival time was measured, as were blood flow and vascular resistance in kidney, intestine, brain, and heart, using the microsphere method. Rats subjected to only myoglobin or hemorrhage survived a period of >120 min; in contrast, rats, exposed to both myoglobin and hemorrhage died at 68 +/- 9 min. When the animals subjected to only hemorrhage and to myoglobin/ hemorrhage were compared, significantly lower values were found in the latter group with respect to blood flow in the kidney (1.7 +/- 0.1 vs. 0.2 +/- 0.05 ml x min(-1) x g(-1)), small intestine (1.0 +/- 0.1 vs. 0.5 +/- 0.1 ml x min(-1) x g(-1)), cardiac output (112 +/- 5 vs. 62 +/- 10 ml(-1) x min(-1) x kg(-1)), and significantly higher values of total peripheral vascular resistance (0.45 +/- 0.02 vs. 0.81 +/- 0.12 mmHg x min x ml( 1) x kg) at 50 min of hypotension. It is assumed that these effects of myoglobin are induced by its ability to scavenge endogenous nitric oxide, because a modified, non-nitrosylable myoglobin was unable to induce such effects. The results support the view that a pathological release of myoglobin into the general circulation causes increases in vascular resistance of vital organs that may contribute to decompensation of tissue supply when occurring in hypovolemic shock. PMID- 12558148 TI - Constitutive nitric oxide synthase inhibition combined with histamine and serotonin receptor blockade improves the initial ovalbumin-induced arterial hypotension but decreases the survival time in brown norway rats anaphylactic shock. AB - Anaphylactic shock accidents after allergen exposure are frequent. After immunization with ovalbumin (OVA), a common dietary constituent, we evaluated the efficacy of pretreatment with histamine-receptor or serotonin-receptor blockers administered alone or in combination with a nitric oxide synthase inhibitor (L NAME) on OVA-induced anaphylactic shock in Brown Norway rats. Animals were allocated to the following groups (n = 6 each): control (0.9% saline); diphenydramine (15 mg kg(-1)); cimetidine (20 mg kg(-1)); diphenydramine + cimetidine; dihydroergotamine (50 microg kg(-1)); diphenydramine + cimetidine + dihydroergotamine; L-NAME (100 mg/kg) alone or associated with diphenydramine, cimetidine, diphenydramine + cimetidine, dihydroergotamine, or diphenydramine + cimetidine + dihydroergotamine. Mean arterial blood pressure (MABP), heart rate (HR), and survival time were monitored for 60 min following treatment. The shock was initiated with i.v. OVA. The MABP drop after i.v. OVA was worsened by diphenydramine and was modestly attenuated by cimetidine, dihydroergotamine, or both together. L-NAME potentiated slightly the effects of cimetidine and dihydroergotamine by lessening the initial MABP decrease, but this transient effect was not sufficient to prevent the final collapse or to improve survival time. Decreased vasodilatory (prostaglandins E2), increased vasoconstrictory (thromboxane B2) prostaglandins, and unchanged leukotriene C4 concentrations were contributory to the overall hemodynamic changes. Thus, the combined blockade of vasodilator mediators (histamine, serotonin, and nitric oxide) slowed the MABP drop in anaphylactic shock, but did not improve survival. More studies are needed to understand these discordant effects. PMID- 12558150 TI - Opioid receptor antagonists modulate Ca2+-activated K+ channels in mesenteric arterial smooth muscle cells of rats in hemorrhagic shock. AB - Previous study has indicated a significant enhancement of activity of large conductance Ca2+-activated K+ channel (BKCa) in mesenteric arterial vascular smooth muscle cells isolated from rats in vascular hyporesponsive stage of hemorrhagic shock. In the present study, the effect of opioid receptor antagonism on BKCa activity in the vascular smooth muscle cells of rats in the hyporesponse stage of hemorrhagic shock was investigated by using inside-out configuration of the patch-clamp technique. The results showed that naloxone (10 microM) down regulated the activity of BKCa by reducing open probability (Po) and open frequency of the channels. The reduction of Po resulted from a decrease of mean open time and an increase of the slow closed time constant. Naltrindole and nor binaltorphimine (100 nM) had the similar effects to that of naloxone, but no significant effect of beta-funaltrexamine (100 nM) on the activity of the channels could be found. These results suggest that delta- and kappa-opioid receptors, but not mu-receptors, may be involved in the regulation of BKCa in vascular hyporesponse stage, and that inhibition of BKCa may be one of the mechanisms of the opioid receptor antagonists improving the response of resistance arteries to vasoactive stimulants during the decompensatory stage of hemorrhagic shock. PMID- 12558151 TI - Diferuloylmethane inhibits neutrophil infiltration and improves survival of mice in high-dose endotoxin shock. AB - Gram-negative septic shock is a systemic inflammatory response of the body caused primarily by the cell wall component (lipopolysaccharide) of the gram-negative bacteria. During high-dose endotoxin shock, neutrophils infiltrate and accumulate in the liver, causing hepatocellular injury. Cell adhesion molecules, specifically intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), play an important role in the infiltration of neutrophils in the liver tissue. In this study, we demonstrate that diferuloylmethane exerts protective effect in high-dose endotoxin shock by improving survival and reducing the severity of endotoxin shock symptoms such as lethargy, diarrhea, and watery eyes following a challenge with lipopolysaccharide. We demonstrate here that diferuloylmethane inhibits the transmigration and infiltration of neutrophils from blood vessels to the underlying liver tissue and, hence, inhibits the damage to the tissue. Diferuloylmethane blocks the induced expression of ICAM-1 and VCAM-1 in liver and lungs. Diferuloylmethane, being a natural compound, may have few side effects and may be useful in attenuating multiple organ injury in pathological conditions arising due to excessive infiltration of neutrophils into the tissues. PMID- 12558152 TI - Specific responses of bacterial cells to dioxins. AB - Five different recombinant bioluminescent strains of Escherichia coli that contain the recA (responsive to DNA damage related stress), fabA (membrane damage), katG (oxidative damage), grpE (protein damage), and lac (constitutive expression, general toxicity) promoters fused to the bacterial lux operon from either Vibrio fischeri or Photorhabdus luminescens were used to describe the different mechanisms of toxicity that several dibenzo-p-dioxins and dibenzofurans have on bacteria, as well as to determine whether bacteria can sensitively detect the presence of these compounds. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (2,3,7,8 TCDD) was found to cause only DNA-related damage to bacterial cells. However, the four stress-responsive strains showed positive responses after addition of 1,2,3,4-tetrachlorodibenzo-p-dioxin (1,2,3,4-TCDD), whereas 2,3,7,8 tetrachlorodibenzo-p-furan (2,3,7,8-TCDF) caused only DNA. oxidative, and protein damage. However, 2,8-dichlorodibenzo-p-dioxin (2,8-DCDD) was not found to induce any stresses tested for in this study, that is, DNA, membrane, oxidative, and protein damage, indicating that each congener might differentially interact with the cell, stimulating differential stress responses within them. By using the constitutive strain, we found that the level of cellular toxicity experienced due to the addition of these four dioxins decreased in the order of 2,3,7,8-TCDD (the most toxic). 1,2,3,4-TCDD, 2.8-DCDD, and 2,3,7,8-TCDF. The 20% effective concentration (EC20), defined in this study the concentration of chemical that causes a 20% decrease in the bioluminescence 60 min after induction, was only 0.1 microg/L for 2,3,7,8-TCDD, a value that is lower than that of the other congeners and demonstrates that 2,3,7,8-TCDD was the most toxic compound tested in this study. PMID- 12558153 TI - The presence of morphologically intermediate papilla syndrome in United Kingdom populations of sand goby (Pomatoschistus spp): endocrine disruption? AB - The sand goby (Pomatoschistus spp.) is a small estuarine fish. Its abundance, life history, and sedentary nature lead to its adoption as a key species in the U.K. Endocrine Disruption in the Marine Environment (EDMAR) Program. This study investigated the presence of classic markers of estrogenic exposure by determining vitellogenin (VTG) and zona radiata protein (ZRP) mRNA levels and ovotestis in estuarine-caught male gobies and investigated morphological changes in the urogenital papilla (UGP). Laboratory exposures to estrogens were also conducted to ascertain the responses of these markers. Wild-caught male fish showed no evidence of ovotestis, VTG, or ZRP mRNA induction. Laboratory exposures suggested that sensitivity of the goby to VTG/ ZRP mRNA induction was similar to flounder. The UGP inspection of wild-caught specimens revealed evidence of feminization of male papillae, a condition denoted as morphologically intermediate papilla syndrome (MIPS). Morphologically intermediate papilla syndrome was more prevalent at estrogenically contaminated sites. Juvenile goby experimentally exposed to 17beta-estradiol for 11 to 32 weeks exhibited signs of the MIPS condition, showing that it was inducible by estrogenic exposure and could therefore be a form of estrogenic endocrine disruption. The estuaries where the MIPS condition was most prevalent (>50% at certain sites) were the Tees, Mersey, and Clyde. The potential of the MIPS condition to significantly interfere with reproductive performance is discussed as well as its use as a monitoring tool for endocrine disruption in the estuarine environment. PMID- 12558154 TI - Toxicity of lead in aqueous medium to desulfovibrio desulfuricans G20. AB - The toxicity of Pb(II) to sulfate-reducing bacteria (SRB) was studied using Desulfovibrio desulfuricans G20 in a medium specifically designed to assess metal toxicity. The effects of Pb(II) toxicity were observed in terms of longer lag times, lower specific growth rates, and in some cases no measurable growth. With an increase in medium pH from 6 to 8, Pb(II) toxicity decreased. At all pH values, in the presence of Pb(II) concentrations ranging from 3 to 15 microM, specific growth rates decreased and lag times increased. The minimum inhibiting concentration (MIC) of Pb(II) causing a complete inhibition in growth at pH 6 was 10 microM, as compared to 15 microM at pH 7.2 and 8. These MIC values are 40 times lower than previously reported for SRB. Results also show that with increases in initial cell protein concentration (inoculum size), soluble Pb(II) removal rates increased and the degree to which Pb(II) caused increased lag times was reduced. In the presence of Pb(II), in all cases in which D. desulfuricans grew (even after a 312-h lag time), the final cell protein concentration was equivalent to that of the Pb-free control. Live/dead staining, based on membrane integrity, indicated that while Pb(II) inhibited growth, Pb(II) did not cause a loss of D. desulfuricans membrane integrity. PMID- 12558155 TI - Abiotic transformation of chlorpyrifos to chlorpyrifos oxon in chlorinated water. AB - In vivo transformation of chlorpyrifos to chlorpyrifos oxon is believed to be a prerequisite for this insecticide to display acute toxicity to organisms. We discovered that active chlorine dispersed in water causes the rapid abiotic transformation of chlorpyrifos to chlorpyrifos oxon. The proposed mechanism for the transformation is an electrophilic attack by hypochlorous acid (HOCl) on the thion (P=S) double bond of chlorpyrifos followed by desulfuration of chlorpyrifos to its oxon analog. The pH of a Cl-aqueous solution controls the speciation of HOCl and hypochlorite (OCl-) in water and thereby influences the transformation process. Chlorpyrifos oxon is a potent anticholinesterase that is about 1,000 times more toxic than chlorpyrifos. Because chlorination is commonly used for treatment of domestic water supplies, the findings raise a new concern about the safety of domestic use of chlorpyrifos products. PMID- 12558156 TI - Adsorption of metals to membrane filters in view of their speciation in nutrient solution. AB - Filtration of a solution may lower metal concentrations through adsorption of metal species to the filter. Processes such as filter-sterilizing nutrient solution and filtration of field water are sensitive to these sorption artifacts, yet basic data on the affinity of different filters for metals are lacking. This article describes the adsorption of five metals to eight types of 0.2-microm membrane filters used for sterilizing a plant (Lemna minor L.) culture medium. Filters of cellulose acetate, cellulose nitrate, mixed cellulose ester, nylon, polyamide, polycarbonate, polyester, and polyvinylidene fluoride were tested for their affinity toward mono- (K), di-(Mn, Cu, Zn), and trivalent (lanthanum [La]) metals. Metal concentrations were quantified using radioisotopes and speciation was calculated. Results showed that metals had the lowest affinity for polycarbonate and nylon filters and the highest affinity for cellulose- and polyester-type filters. Furthermore, it was shown that the metal load on cellulose filters correlated best with free ion concentrations (indicating electrostatic attraction), while loads on other filters correlated better with total metal concentrations. Filtering a 5-ml solution of pH 5 did not affect its metal concentrations, ranging from 10 nM (La) to 49 microM (K). To minimize filtration artifacts, we propose using polycarbonate or nylon filters, especially when dealing with low volumes of high pH and low metal species concentrations. PMID- 12558157 TI - Quantitative structure-biodegradation relationship study for biodegradation rates of substituted benzenes by river bacteria. AB - The biodegradation rate constant (K) of 21 substituted phenols, anilines, and benzoic acids was determined by using bacteria from the Songhua River (Jilin Province, China) as an inoculum. All of the compounds studied are readily biodegradable chemicals, as shown by their high K values. The quantitative structure-biodegradation relationship results showed that the biodegradation rate of the studied compounds is related mainly to the heat of formation (Hf) and the negative logarithm of the ionization constant (pKa), whereas no simple relationship exists with the n-octanol-water partition coefficients. The equation K = - 1.127 x 10(-3)Hf - 0.0388pKa + 0.859 was found to fit well (r2(adj) = 0.805), and the correlation coefficient between the predicted and the experimental K values was up to 0.908. PMID- 12558158 TI - The bioconcentration and metabolism of chlorpyrifos by the eastern oyster, Crassostrea virginica. AB - Eastern oysters (Crassostrea virginica) were exposed to [14C]chlorpyrifos (O,O diethyl-O-[3,5,6-trichloro-2-pyridyl] phosphorothioate) at an average measured seawater concentration of 0.6 microg/L under flow-through conditions for 28 d. The compound O,O-diethyl-O-(3,5-dichloro-6-methylthio-2-pyridyl)phosphorothioate (DMP) was extracted and identified as the single metabolite observed, and this metabolite constituted the majority of the total [14C] activity in the oyster at all sampling times. Once oysters were exposed to clean water, both chlorpyrifos and DMP residues cleared rapidly from whole oysters, with elimination half-lives of <3 d. A simple two-compartment uptake/elimination model was adequate to describe total [14C] activity in whole oysters, edible tissue, and oyster liquor. The average bioconcentration factors (BCFs) for total [14C] activity in whole oysters, edible tissue, and oyster liquor were 565, 1,400, and 35 ml/g, respectively. The parent [14C]chlorpyrifos accumulated to a peak residue concentration of 135 microg/kg in whole oyster tissue, representing an empirical [14C]chlorpyrifos BCF value in the oyster of approximately 225 ml/ g; the BCF value for [14C]chlorpyrifos was lower than the BCF for total [14C] activity in whole oysters and edible tissue because of extensive metabolism to DMP and oyster elimination processes. PMID- 12558159 TI - Immunocompetence of juvenile Chinook salmon against Listonella anguillarum following dietary exposure to Aroclor 1254. AB - Controlled laboratory challenges with pathogenic Listonella (formerly Vibrio) anguillarum bacteria were used to examine potential effects of dietary exposure to polychlorinated biphenyls (PCBs) on the growth and immunocompetence of juvenile Puget Sound (WA, USA) Chinook salmon (Oncorhynchus tschawytscha). Salmon were fed four levels of the PCB congener mixture Aroclor 1254 for 28 d to bracket likely exposure to PCBs in the lower Duwamish waterway near Seattle, Washington, USA. Fish were transferred to five replicate tanks per dose, exposed to L. anguillarum, and monitored for 14 d. Half the PCB-dosed fish were vaccinated against L. anguillarum, and specific immunity was allowed to develop in this group for three weeks prior to challenge. All mortalities following challenge were individually sampled for bacteria to identify the cause of death. The data indicate that dietary PCB exposure, even at relatively high levels, did not have a significant effect on growth, innate disease resistance, or acquired immunity to L anguillarum. The controlled laboratory experiments in this study suggest that the immune system of Chinook salmon is not sensitive to orally delivered PCBs at environmentally relevant concentrations. PMID- 12558160 TI - Evaluating reptile exposure to cholinesterase-inhibiting agrochemicals by serum butyrylcholinesterase activity. AB - Blood samples from lizards (Gallotia galloti) collected from two agricultural areas (Las Galletas and Punta del Hidalgo) and two reference areas on the Island of Tenerife (Canary Islands, Spain) were analyzed for butyrylcholinesterase (BChE) activity. Serum BChE activity was characterized first by in vitro experiments using selective substrates and inhibitors. Of the total cholinesterase (ChE) activity, 74% could be attributed to BChE activity. This portion of the total ChE activity was inhibited dose dependently by tetraisopropyl pyrophosphoramide and hydrolyzed the substrate butyrylthiocholine iodide. No enzyme inhibition was observed at high substrate concentration. Twenty one lizards collected from agricultural sampling sites showed significant inhibition (p < 0.001) of BChE activity (mean +/- standard deviation [SD] of 4.66 +/- 2.63 micromol/min/ml for lizards from Las Galletas and 5.13 +/- 1.48 for lizards from Punta del Hidalgo) compared with BChE activity for lizards from the reference sites (6.35 +/- 1.75 micromol/min/ ml). Las Galletas had the highest number of lizards (22%) with significantly inhibited BChE activity. In vitro assays showed that 10(-4) M pyridine-2-aldoxime methochloride (2-PAM) reactivated dichlorvos- or paraoxon-inhibited BChE activity within a 60-min incubation period. Almost all serum samples with depressed BChE activity that were collected from lizards from agricultural areas responded to 2-PAM reactivation of enzyme activity (8-60% increase in enzyme activity). Reactivation by treatment with 2 PAM confirmed that the depression of BChE activity was attributable to organophosphorus (OP) compounds. Evaluation of BChE activity levels and the chemical reactivation of serum BChE activity in G. galloti using 2-PAM was found to be a sensitive indicator of G. galloti exposure to OP compounds. PMID- 12558161 TI - Assessing the toxicity of dodecylbenzene sulfonate to the midge Chironomus riparius using body residues as the dose metric. AB - Dodecylbenzene sulfonate (DBS) is a component of linear alkylbenzene sulfonate (LAS), an anionic surfactant, mainly used in household detergents. Due to the large quantity of DBS in use, there is concern over adverse environmental effects. This work examined the toxicokinetics and toxicity of the 2-phenyl isomer of dodecylbenzene sulfonate in 4-d, 10-d, and partial life-cycle tests on the midge, Chironomus riparius, exposed to aqueous solutions. Toxicokinetics were determined in 10-d uptake and 5-d elimination tests. The toxicokinetics were based on parent compound concentration in water and yielded an uptake coefficient (ku) of 17.5 (14.87-20.20) ml/g/h, an elimination rate constant (ke) of 0.073 (0.062-0.085) per h, a bioconcentration factor (BCF) of 56 to 240, and a half life (t 1/2) of 9.5 (8.0-11.0) h. Biotransformation measurements did not reveal evidence for DBS metabolism. Thus, body residues, determined in the toxicity study, represent parent compound. In toxicity tests, 4- and 10-d LR50s (the body residue required to cause 50% mortality) in live midges were 0.72 (0.65-0.79) and 0.18 (0.08-0.42) mmol/kg, respectively. Thirty-day LR50s were 0.18 (0.09-1.64) and 0.21 (0.15-0.39) mmol/kg in duplicate studies. Of the sublethal endpoints, only developmental time increase was significant, with the lowest-observed-effect residues of 0.085 (0.067-0.105) and 0.100 (0.087-0.114) mmol/kg for male and female midges, respectively. Deformities in surviving larvae were also observed as chronic responses for body residues exceeding the 30-d LR50. The body residues required for mortality suggest that DBS acts like a polar narcotic in the midge. PMID- 12558162 TI - Responses to selection for cadmium resistance in the least killifish, Heterandria formosa. AB - An artificial selection experiment was conducted for six generations to investigate the responses to selection for cadmium resistance in the least killifish, Heterandria formosa. There was a rapid response to selection. After only one generation of selection, two of the three selection lines had an increased resistance to cadmium, while all three selection lines had an elevated resistance by the next generation. After six generations of selection, fish from the selection lines survived about three times as long as control line fish when exposed to cadmium. Realized heritability for cadmium resistance was estimated to be 0.50. Cross-resistance to copper was studied in the F2, F3, and F6 generations; fish from the cadmium-resistant lines had a longer survival time than those from the control lines. Resistance to heat was investigated in the F2, F3, and F5 generations; the cadmium-resistant lines had a significantly shorter survival time than the control lines. Our results showed that evolution of resistance to cadmium was rapid, that cross-resistance to copper occurred, and that the evolution of resistance to cadmium was accompanied by a trade-off (fitness costs) as evidenced by the reduced heat tolerance. PMID- 12558163 TI - Exposure to the polychlorinated biphenyl mixture Aroclor 1254 alters melanocyte and tail muscle morphology in developing Xenopus laevis tadpoles. AB - Polychlorinated biphenyls (PCBs) are ubiquitous environmental contaminants that have damaging effects on both ecosystem and human health. Numerous studies have shown that exposure to PCBs can alter growth and development of aquatic organisms, including frogs. In this report, developing Xenopus laevis tadpoles were exposed to the PCB mixture Aroclor 1254. Tadpoles were exposed from 5 through 9 d postfertilization to either 0, 1, 10, 50, or 100 ppm Aroclor 1254. Exposure to an acute, high concentration of Aroclor 1254 (10, 50, and 100 ppm) caused statistically significant reductions in survival and body size. In addition, tadpoles exposed to these higher concentrations showed histological abnormalities, including aberrant tail tip, myotomal, and melanocyte morphologies. Described adverse health effects associated with PCB exposure of developing frogs will serve as useful health endpoints in ongoing and future molecular-based studies that correlate health effects with changes in gene expression. PMID- 12558164 TI - In vitro and in vivo estrogenicity and toxicity of o-, m-, and p-dichlorobenzene. AB - The estrogenicity of o-, m-, and p-dichlorobenzene (DCB) was evaluated with a yeast estrogen screen (YES) and zebrafish (Danio rerio) vitellogenin (VTG) assays. With the YES, p-DCB and m-DCB were found to be estrogenic in a concentration-responsive manner. The relative potency measured with the YES (relative to 17beta-estradiol) was 2.2 x 10(-7) for p-DCB and 1.04 x 10(-8) for m DCB. Following acute toxicity tests with the zebrafish, plasma VTG production was measured to examine the in vivo estrogenic activity of the three compounds after a 14-d exposure. Adult zebrafish were exposed to different concentrations of o-, m- and p-DCB, ranging from 0.1 to 32 mg/L; ethynylestradiol ([EE2]; 5 ng/L, 10 ng/L, 50 ng/L, and 100 ng/L) was used as a positive control. After exposure, blood samples were taken and protein electrophoresis was performed to determine the relative VTG content. Gonadosomatic indices (GSI) and condition factors (CF) were also calculated. Elevated VTG levels and decreased female GSIs were found in fish exposed to > or = 5 ng EE2/L and in fish exposed to > or = 10 mg p-DCB/L. Low GSIs coincided with high levels of VTG in the blood of female zebrafish. This relation was not only found in fish exposed to EE2 but also in controls and fish exposed to DCB. Therefore, a direct or indirect effect of VTG on the GSI is suggested rather than a direct toxic effect of the tested compounds on the gonads. PMID- 12558165 TI - Joint acute toxicity of esfenvalerate and diazinon to larval fathead minnows (Pimephales promelas). AB - California (USA) agriculture employs pyrethroid and organophosphate insecticides to control insects in orchards and other crops. Diazinon and esfenvalerate were selected for this study because of their application overlaps. Toxicological and biochemical responses of larval fathead minnows (Pimephales promelas) exposed singly and in combinations to esfenvalerate and diazinon were determined. Exposures were 96-h static renewal tests that used standard U.S. Environmental Protection Agency acute toxicity test methods. After pesticide exposures, larvae were evaluated for carboxylesterase and acetylcholinesterase activity, and histopathological effects. Carboxylesterase activity was examined because of its potential influence on the toxicity of both organophosphates and pyrethroids. In vivo studies demonstrated that diazinon significantly inhibited carboxylesterase activity at nominal water concentrations as low as 50 microg/L. However, esfenvalerate did not affect carboxylesterase activity at any concentration tested. Liver glycogen depletion was the only histopathological effect observed; this effect was demonstrated with the individual pesticides and pesticide combinations (i.e., mixtures). The combinations of diazinon and esfenvalerate causing acute toxicity to fathead minnow larvae appeared to be greater than additive (i.e., synergistic) in all three tests. PMID- 12558166 TI - Influence of dietary sodium on waterborne copper toxicity in rainbow trout, Oncorhynchus mykiss. AB - Juvenile rainbow trout were fed diets containing control (0.26 mmol/g) or elevated (1.3 mmol/g) dietary Na+ in combination with either background (19 nmol/L) or moderately elevated levels (55 or 118 nmol/L) of waterborne Cu for 21 d. Unidirectional waterborne Na+ uptake rates (measured with 22Na) were up to four orders of magnitude higher than those of Cu (measured with 64Cu). Chronic exposure to elevated dietary Na+ alone or in combination with elevated waterborne Cu decreased whole-body uptake rates of waterborne Na+ and Cu. Accumulation of new Cu and Na+ at the gills was positively and highly significantly correlated and responded to the experimental treatments in a similar fashion, suggesting that Na+ and Cu have common branchial uptake pathways and that dietary Na+ preexposure modifies these pathways. Chronic exposure to elevated waterborne Cu significantly increased Cu concentrations in the liver but caused only modest increases in total Cu concentrations in the whole body and gill. Chronic exposure to elevated dietary Na+ slightly decreased whole-body Cu concentration on day 14 and greatly reduced liver Cu concentration on days 14 and 21; new Cu accumulation in whole-body, gill, and internal organs was reduced on all days. Chronic exposure to elevated waterborne Cu or dietary Na+ alone reduced short-term gill Cu binding at low waterborne Cu concentrations. At high waterborne Cu concentrations, chronic exposure to elevated waterborne Cu had no effect, while elevated dietary Na+ increased Cu binding to the gills. Combined chronic exposure to elevated dietary Na+ and waterborne Cu decreased gill Cu binding over the entire range of Cu concentrations tested. Clearly, chronic exposure to elevated dietary Na+ and waterborne Cu appears to modify gill Cu-binding characteristics and may be important considerations in future development of a chronic biotic ligand model for Cu. PMID- 12558167 TI - Factors influencing the design of bioaccumulation factor and biota-sediment accumulation factor field studies. AB - A series of modeling simulations was performed to evaluate the underlying factors and principles that drive the uncertainty in measured bioaccumulation factors (BAFs) and biota-sediment accumulation factors (BSAFs) for fish, and to determine which sampling designs minimize those uncertainties. Temporal variability of chemical concentrations in the water column, and the metabolism rate and n octanol-water partition coefficient (Kow) for the chemical were determined to be the dominant factors that influenced the field-sampling design. Spatial variability of the chemical concentrations, food web structure, and the sediment water column concentration quotient had a lesser importance upon the overall design. The simulations also demonstrated that collection of composite water samples in comparison to grab water samples resulted in reductions in the uncertainties associated with measured BAFs. Some illustrative sampling design structures for BAF and BSAF measurements based upon the temporal variability of chemical concentrations in the water column and the metabolism rate and Kow for the chemical were developed. These illustrative designs provided a sense of how sampling design structures, that is, the number and spacing over time of sampling events, might be influenced by differences in temporal variabilities, metabolism rates, and Kow. Although the importance of spatial variability was discounted as a major factor in the design process, sampling of water and sediment across the immediate home range of the sampled organism is required for successful measurements because poor spatial coordination of organism, water, or sediment samples will yield BAFs and BSAFs with large and unknown biases. PMID- 12558168 TI - Laboratory and field validation of multiple molecular biomarkers of contaminant exposure in rainbow trout (Oncorhynchus mykiss). AB - Novel primers for three different genes (CYP1A1, vitellogenin, and metallothionein) for use in reverse transcription-polymerase chain reaction (RT PCR) were developed and tested in laboratory flow-through experiments using model compounds benzo[a]pyrene, estradiol, and cadmium. Field validations of the molecular biomarkers were performed using 8-d or 48-h in situ, cagedexposure of juvenile rainbow trout (Oncorhynchus mykiss). Expression levels of genetranscription(messengerRNA [mRNA]) were monitored in gill and liver tissue after exposure at sites downstream of a known source of creosote contamination in the Little Scioto River (Marion, OH, USA) and metals and hydrocarbon contamination in Dick's Creek (Middletown, OH, USA). Significant increases of liver cytochrome P4501A1 gene (CYP1A1) and vitellogenin were measured in both streams at sites downstream of contaminant sources. Significant increases of gill CYP1A1 were also observed. Significant increases of gill and livermetallothionein occurred in Dick's Creek. A sample of gill tissue taken from each fish prior to placement in the Little Scioto River provided a pre-exposure control measurement for each individual. Expression of CYP1A1 and vitellogenin in trout at both streams indicated that exposure to bioavailable hydrocarbons and endocrine disrupting compounds had taken place. Previous to this study, estrogenic compounds had not been detected in the Little Scioto River. Metallothionein expression confirmed the presence of metals in Dick's Creek. These results suggest a mixed contaminant environment that poses physiological hazard to existing teleost populations. Multiple molecular biomarkers provided a sensitive, physiologically based method of determining contaminant presence and bioavailability to aquatic organisms. PMID- 12558169 TI - Associations between regional differences in polychlorinated biphenyls and dichlorodiphenyldichloroethylene in blood of nestling bald eagles and reproductive productivity. AB - The relationship between regional reproduction rates of bald eagles (Haliaeetus leucocephalus) and concentrations of p.p'-dichlorodiphenyldichloroethylene (p,p' DDE) and total polychlorinated biphenyls (PCBs) in blood plasma from nestling bald eagles was assessed. Blood was analyzed from 309 nestlings from 10 subpopulations of eagles across the Great Lakes region. Geometric mean concentrations of p,p'-DDE and total PCBs were inversely correlated to the productivity and success rates of nesting bald eagles within nine subpopulations. Nestlings eight weeks of age and older had significantly greater geometric mean concentrations of total PCBs and p,p'-DDE than nestlings less than eight weeks of age. The ability to use measurements of p,p'-DDE and total PCBs in nestling blood to determine the potential impact of these contaminants on adult nesting on a regional scale was demonstrated. PMID- 12558170 TI - The effects of chronic cadmium exposure on American toad (Bufo americanus) tadpoles. AB - American toad (Bufo americanus) tadpoles were exposed to cadmium from shortly after hatching through metamorphosis. Cadmium at 540 microg/L decreased percentage survival and metamorphosis; 5 and 54 microg/L increased growth relative to controls. Amphibians exposed to cadmium as tadpoles may be less tolerant of natural stressors in the terrestrial environment. PMID- 12558171 TI - Dietary oral exposure to l,3,5-trinitro-1,3,5-triazine in the northern bobwhite (Colinus virginianus). AB - The potential risk to wildlife from exposure to explosives, including 1,3,5 trinitro-1,3,5-triazine (RDX), has been an issue at numerous U.S. military installations where these substances are found in soil and water. Presently, no data describing the effects of RDX exposure in avian species exist. Therefore, an acute lethal dose (ALD) and 14- and 90-d subchronic dietary exposures to RDX were evaluated in a species potentially present at many contaminated sites, i.e., the northern bobwhite (Colinus virginianus). The ALDs for females and males were 187 and 280 mg/kg, respectively. Data from the 14-d dietary trial suggested that RDX exposure inhibited food consumption, weight gain, and egg production. Dietary RDX exposure for 90-d produced a dose-dependant decreasing trend in total feed consumption, total egg production, and hen-housed production parameters. These collective data suggest that quail may respond differently to oral RDX exposure compared with mammals. PMID- 12558172 TI - Comparison of metal accumulation in mussels at different local and global scales. AB - Cadmium and zinc uptake from the dissolved phase, assimilation efficiency (AE) from the dietary phase, and body burden as well as clearance rate were measured in green mussels, Perna viridis, and blue mussels Mytilus edulis, M. galloprovincialis and Mytilus trossulus. Perna viridis was collected from four sites differentially enriched with trace metals in Hong Kong and blue mussels were collected from different climatic zones, i.e., subarctic and temperate, to allow comparisons with the more tropical green mussels. Despite similar shell length, the dry weight of mussels varied significantly between sites and species and this had a large effect on Cd and Zn accumulation, clearance rate, and metal body burden. All data were, therefore, weight adjusted to allow comparison without this confounding factor. Trace-metal body concentrations were significantly different between sites, and P. viridis collected from Tsing Yi, Hong Kong, had the highest levels of all measured metals when compared with other Hong Kong sites. There was, however, no relationship between the degree of metal enrichment and the Cd and Zn uptake (both from dissolved and particulate sources) and clearance rates. Furthermore, Cd and Zn uptake (dissolved and particulate) and clearance rate varied little between species or climatic zones of collection. Thus, over the range of body trace-metal concentrations measured and between mussel species over large geographical distances and climatic zones, the uptake rates, AEs, and clearance rates are similar when measured under the same laboratory conditions after body-size correction. When other factors such as salinity are also corrected, biomonitoring data from different areas and even utilizing different mussel species may be directly comparable. This study therefore provides important evidence in support of Mussel Watch Programs. PMID- 12558173 TI - Response of larval Xenopus laevis to atrazine: assessment of growth, metamorphosis, and gonadal and laryngeal morphology. AB - Larval Xenopus laevis were exposed to one of four concentrations of atrazine (0, 1, 10, or 25 microg/L, 11 replicate tanks per treatment, 60-65 larvae per replicate) dissolved in an artificial pond water (frog embryo teratogenesis assay Xenopus [FETAX]) medium beginning 48 h after hatching until the completion of metamorphosis. Separate groups of larvae (six replicate tanks per treatment, 60 65 larvae per replicate) were exposed to estradiol (100 microg/L), dihydrotestosterone (100 microg/L), or ethanol vehicle control dissolved in FETAX medium. None of the treatments affected posthatch mortality, larval growth, or metamorphosis. There were no treatment effects on sex ratios except for estradiol, which produced a greater percentage of female offspring. Exposure to either estradiol or 25 microg atrazine/L increased the incidence of intersex animals based on assessment of gonadal morphology. Atrazine did not reduce the size of the laryngeal dilator muscle, a sexually dimorphic muscle in this species. We conclude that environmentally relevant concentrations of atrazine do not influence metamorphosis or sex ratios and do not inhibit sexually dimorphic larynx growth in X. laevis. The incidence of atrazine-induced intersex animals was small (<5%) and occurred only at the greatest concentration of atrazine tested, a concentration that is rarely observed in surface waters in the United States. PMID- 12558174 TI - Environmental exposure to polychlorinated biphenyls among raccoons (Procyon lotor) at the paducah gaseous diffusion plant, Western Kentucky, USA. AB - An investigation involving raccoons (Procyon lotor) as a sentinel species at the Paducah Gaseous Diffusion Plant (PGDP) in Western Kentucky (USA) delineated the extent of exposure to polychlorinated biphenyls (PCBs) and PCB spatial distribution. Raccoon exposure to PCBs was demonstrated through analysis of subcutaneous fat, abdominal fat, liver, and brain tissues from raccoons collected at the PGDP but also was clearly evident in raccoons from a reference area situated along the Ohio River (USA). Raccoons with the highest tissue PCB concentrations appeared to be those inhabiting areas nearest the plant itself and most likely those that ventured into the plants interior. Male raccoons at the PGDP had similar concentrations of total PCBs in subcutaneous fat (1.86 +/- 0.64 microg/g) as males from the reference site (1.41 +/- 0.35 microg/g), but females had higher PCB body burdens than those at the reference site (9.90 +/- 6.13 microg/g vs 0.75 +/- 0.40 microg/g). Gross measurements of exposure to radiation producing materials revealed that counts per minute exceeded background in 61% of PGDP raccoons compared with 27% at the reference site and five raccoons at the PGDP had beta counts that were more than twice the background. Differences among trapping success, growth rates, and serum chemistry parameters were noted but may have been related to habitat and other environmental and population density factors. PMID- 12558175 TI - Environmental polychlorinated biphenyl exposure and cytochromes P450 in raccoons (Procyon lotor). AB - An investigation involving raccoons as a sentinel species at the Paducah Gaseous Diffusion Plant (PGDP) and Ballard Wildlife Management Area in western Kentucky (USA) delineated the extent of exposure to polychlorinated biphenyls (PCBs). Three separate measures of hepatic cytochrome P450 (CYP) induction were used to evaluate raccoon physiological responses to PCB exposure. Hepatic CYP induction was estimated via determination of total CYP, dealkylase activities, and immunoreactive proteins. There were no differences in raccoon biomarker responses between study sites. Significant relationships between and among PCB residues and biomarkers indicated that hepatic CYP induction had occurred in response to PCB exposure. Pentoxyresorufin O-deethylase (PROD) activity, CYP1A1, and CYP1A2 were biomarkers most closely associated with PCB exposure. The rank order of responses was CYP1A1 > CYP1A2 > PROD > ethoxyresorufin O-deethylase (EROD) as related to raccoon liver PCB concentrations, whereas the order was CYP1A1 > PROD > EROD > CYP1A2 when regressed with total PCB concentrations in abdominal fat. PMID- 12558176 TI - Disturbances to metal partitioning during toxicity testing of iron(II)-rich estuarine pore waters and whole sediments. AB - Metal partitioning is altered when suboxic estuarine sediments containing Fe(II) rich pore waters are disturbed during collection, preparation, and toxicity testing. Experiments with model Fe(II)-rich pore waters demonstrated the rates at which adsorptive losses of Cd, Cu, Ni, Mn, Pb, and Zn occur upon exposure to air. Experiments with Zn-contaminated estuarine sediments demonstrated large and often unpredictable changes to metal partitioning during sediment storage, removal of organisms, and homogenization before testing. Small modifications to conditions, such as aeration of overlying waters, caused large changes to the metal partitioning. Disturbances caused by sediment collection required many weeks for reestablishment of equilibrium. Bioturbation by benthic organisms led to oxidation of pore-water Fe(II) and lower Zn fluxes because of the formation of Fe hydroxide precipitates that adsorb pore-water Zn. For five weeks after the addition of organisms to sediments, Zn fluxes increased slowly as the organisms established themselves in the sediments, indicating that the establishment of equilibrium was not rapid. The results are discussed in terms of the dynamic nature of suboxic, Fe(II)-rich estuarine sediments, how organisms perturb their environment, and the importance of understanding chemistry in toxicity testing with whole sediments or pore water. Recommendations are provided for the handling of sediments for toxicity testing. PMID- 12558177 TI - Sublethal energetic responses by Pisidium amnicum (Bivalvia) exposed to pentachlorophenol at two temperatures. AB - Sublethal effects of pentachlorophenol (PCP) were studied in the freshwater clam Pisidium amnicum by examining the cause-effect relationship between PCP body burden and the rate of the whole animal heat output. In addition, the modifying effects of temperature (7 and 15 degrees C) and trematode parasites as natural stress factors were assessed. Results showed that even a low PCP body burden was sufficient to increase the heat output during the periods of aerobic metabolism and the magnitude of responses increased linearly with increasing body burden. During a valve-closure period, however, PCP had no effect on anaerobic heat output. The rise of the maximum heat output in response to PCP was more pronounced at the higher temperature. The response in heat output was similar in both infected and uninfected clams, but the general level of heat output was lower in infected clams. The increasing PCP body burden also increased the duration of valve-closure time. The results of this study suggested that microcalorimetry could be a useful tool in assessing sublethal responses by the clam, which offers several calorimetrically measurable parameters such as aerobic and anaerobic heat output and behavior. PMID- 12558178 TI - Toxicokinetics of sediment-sorbed benzo[a]pyrene and hexachlorobiphenyl using the freshwater invertebrates Hyalella azteca, Chironomus tentans, and Lumbriculus variegatus. AB - This study investigated the effect of long-term sediment aging on the toxicokinetics of benzo[a]pyrene (BaP) and hexachlorobiphenyl (HCBP) using three freshwater benthic invertebrates. Hyalella azteca, Chironomus tentans, and Lumbriculus variegatus were exposed to BaP- and HCBP-spiked sediments that were aged for 7 d or 1.5 years. The toxicokinetics of the two compounds were determined for each test organism using a two-compartment model. The modeling of BaP was more complex because biotransformation was included within the model. The results of this study showed that the HCBP uptake clearance rates (k(s)) for each species were generally an order of magnitude greater than those determined for BaP and this difference was most likely due to preferential and rapid binding of BaP to sediment particles. Overall, the bioavailability of HCBP in spiked sediments tended to decrease with duration of aging, based on k(s) values and bioaccumulation factors (BAFs). However, the decreases in bioavailability appear to be species specific. Benzo[a]pyrene did not decline in bioavailability for the species tested because it may resist movement into the micropores of the sediment due to its large size. In addition to the bioassays, this article outlines a method for toxicokinetic modeling of biotransformed compounds and methods for statistical comparisons of kinetic parameters (i.e., k(s), k(d)...) and BAF values. PMID- 12558179 TI - Adsorption, complexation, and phytoavailability of copper as influenced by organic manure. AB - Copper (Cu) is bound strongly to clay minerals and organic matter in soils, and forms both insoluble and soluble organic complexes with organic carbon. In this experiment, the effect of five manure composts (biosolid, farmyard manure, spent mushroom, pig manure, and poultry manure) on the adsorption and complexation of Cu in a mineral soil (Manawatu sandy soil, Palmerston North, New Zealand) low in organic matter content was examined. The effect of biosolid on the uptake of Cu from the soil, treated with various levels of Cu (0-400 mg/kg soil), was examined by using mustard (Brassica juncea L.) plants. The redistribution of the added Cu in soil was evaluated by a chemical fractionation scheme. Addition of manure compost increased the adsorption and complexation of Cu by the soil. At the same level of total organic carbon addition, a significant difference was found in the extent of Cu adsorption among the manure-amended soils. However, less difference was found in the amount of Cu complexed among the manure-amended soils. A significant inverse relationship was found between the extent of Cu adsorption and the dissolved organic carbon (DOC) in the manure-amended samples, indicating that DOC formed soluble complexes with Cu. Increasing addition of Cu increased Cu concentration in plants, resulting in decreased plant growth at high levels of Cu (i.e., phytotoxicity). Addition of biosolid was found to be effective in reducing the phytotoxicity of Cu at high levels of Cu addition. Significant relationships were found between dry matter yield and total Cu or free Cu2+ concentration in soil solution. Addition of biosolid decreased the concentration of the soluble and exchangeable Cu fraction but increased the concentration of the organic-bound Cu fraction in soil. PMID- 12558180 TI - Anaerobic benzene biodegradation--a new era. AB - Benzene is biodegraded in the absence of oxygen under a variety of terminal electron-accepting conditions. However, the mechanism by which anaerobic benzene degradation occurs is unclear. Phenol and benzoate have been consistently detected as intermediates of anaerobic benzene degradation, suggesting that the hydroxylation of benzene to phenol is one of the initial steps in anaerobic benzene degradation. The conversion of phenol to benzoate could then occur by the carboxylation of phenol to form 4-hydroxybenzoate followed by the reductive removal of the hydroxyl group to form benzoate. 13C-Labeling studies suggest that the carboxyl carbon of benzoate is derived from one of the carbons of benzene. Although the fumarate addition reaction is commonly used to activate many hydrocarbons for anaerobic degradation, the large activation energy required to remove hydrogen from the benzene ring argues against such an approach for anaerobic benzene metabolism. The alkylation of benzene to toluene has been detected in several mammalian tissues, and offers an interesting alternate hypothesis for anaerobic benzene degradation in microbial systems. In support of this, anaerobic benzene degradation by Dechloromonas strain RCB, the only known species to degrade benzene in the absence of oxygen, is stimulated by the addition of vitamin B12 and inhibited by the addition of propyl iodide which is consistent with the involvement of a corrinoid enzymatic step. Alkylation of benzene to toluene is also consistent with labeling data that suggests that the carboxyl carbon of benzoate is derived from one of the benzene carbons. However, it is difficult to envision how phenol would be formed if benzene is alkylated to toluene. As such, it is possible that diverse mechanisms for anaerobic benzene degradation may be operative in different anaerobic microorganisms. PMID- 12558181 TI - Phylogenetic reconstruction within Mycobacterium tuberculosis Beijing genotype in northwestern Russia. AB - A selection of genetic markers was used to study the evolution of Mycobacterium tuberculosis Beijing family strains in northwestern Russia. A total of 221 of 434 epidemiologically unlinked isolates studied in 1996-2001 belonged to the Beijing family as determined by standard spoligotyping (signals 35-43). Ninety-six percent of these Beijing isolates ("typical") were closely related in IS6110-RFLP (D > 0.85) while 9 remaining isolates (2 different profiles, "atypical") were more distant from the rest (D = 0.6-0.7). Further analysis was performed on a selection of 12 typical and both atypical Beijing strains with different IS6110 RFLP profiles (2 isolates each). All 28 Beijing isolates studied had the KatG 463Leu allele, an intact mtp40 fragment of the mpcA gene, and an identical structure of the DR locus (15 DVRs) with an upstream IS6110 copy in opposite orientation. The IS6110-RFLP based neighbor-joining (distance) and quartet puzzling (maximum-likelihood) trees showed that the branch lengths were considerably longer for atypical Beijing strains. Typical Beijing strains had the 1.02 kb Rv3135 PPE-family gene and two IS1547 copies (iplA and iplB) one of them (iplB) disrupted by IS6110 insertion. Atypical Beijing strains had the 1.97 kb Rv3135 gene and a single intact IS1547/iplA copy. We suggest that the M. tuberculosis Beijing family strains currently circulating in the northwest of Russia are relatively ancient and thus appear to be endemic in this region since evolutionarily distant time. The prevalent typical Beijing strains (96%) are likely to be of monophyletic origin and their ongoing dissemination has started recently: these strains differ in rapidly evolving IS6110-RFLP but have identical structure of other polymorphic genome regions studied. The atypical Beijing strains (4%) are evolutionary older; they probably had a common (unknown) ancestor with typical Beijing strains. PMID- 12558182 TI - DcrA and dcrB Escherichia coli genes can control DNA injection by phages specific for BtuB and FhuA receptors. AB - We had previously shown that the Escherichia coli proteins DcrA (SdaC) and DcrB, located, respectively, in the inner membrane and periplasm, are involved in the early development of virulent bacteriophage C1, which recognises BtuB as an outer membrane receptor. In the present work it is demonstrated that the DcrA and DcrB proteins, coordinately with another outer membrane receptor protein, FhuA, are also involved in an early stage of development of a newly isolated virulent phage, C6. In both cases, DcrA and DcrB probably are required in the second stage of phage adsorption-the DNA injection process. This means that DcrA and DcrB proteins can participate in phage DNA transport pathways in cooperation with different outer membrane receptors, including FhuA and BtuB. The increased sensitivity of bacterial cells to SDS following C1 and C6 adsorption suggests that adsorption by these phages triggers the opening of diffusion channels through the outer membrane. Our results also indirectly demonstrate that the DcrA and DcrB proteins participate in the opening or formation of these channels. PMID- 12558183 TI - Antimicrobial and antiviral activities of polyphenolics from Cocos nucifera Linn. (Palmae) husk fiber extract. AB - The decoction of Cocos nucifera L. husk fiber has been used in northeastern Brazil traditional medicine for treatment of diarrhea and arthritis. Water extract obtained from coconut husk fiber and fractions from adsorption chromatography revealed antimicrobial activity against Staphylococcus aureus. The crude extract and one of the fractions rich in catechin also showed inhibitory activity against acyclovir-resistant herpes simplex virus type 1 (HSV-1-ACVr). All fractions were inactive against the fungi Candida albicans, Fonsecaea pedrosoi and Cryptococcus neoformans. Catechin and epicatechin together with condensed tannins (B-type procyanidins) were demonstrated to be the components of the water extract. PMID- 12558184 TI - Presence in bovine enteropathogenic (EPEC) and enterohaemorrhagic (EHEC) Escherichia coli of genes encoding for putative adhesins of human EHEC strains. AB - Enteropathogenic and enterohaemorrhagic Escherichia coli (EPEC and EHEC) infections are characterised by the formation of attaching and effacing lesions on intestinal epithelial cells. The first step of EPEC and EHEC pathogenesis involves the initial adherence of the bacterium to the intestinal epithelium. A collection of bovine EPEC and EHEC strains belonging to different serogroups was tested by colony blot hybridization with gene probes for putative adhesins (BFPA, LPFA, IHA, LIFA) of human EPEC and EHEC, and also for fimbrial and afimbrial adhesins (AFA8, F17, Cs31A) of bovine necrotoxigenic E. coli (NTEC). In the bovine EPEC and EHEC strains tested, sequences homologous to lifA, ihA, and lpfA genes were detected, sometimes in association with particular serogroups. Bovine 026 EPEC also possessed a sequence homologous to a gene of the c/p operon, coding for the CS31A adhesin, associated with bovine NTEC. Overall results showed that different genes encoding for putative adhesins of human EHEC strains are present in bovine EPEC and EHEC strains, but not one of them is present in all strains. PMID- 12558185 TI - Expression of UreI is required for intragastric transit and colonization of gerbil gastric mucosa by Helicobacter pylori. AB - Helicobacter pylori colonizes the antral mucosa of the human stomach. There is a controversy as to whether the microorganism is exposed to acidity in its ecological niche. In vitro, the microorganism requires urease for gastric colonization and survival at pH < 4.0. UreI encodes an acid activated urea channel enabling urea access to intrabacterial urease at acidic pH. UreI is also necessary for survival at pH < 4.0. However, the role of UreI for both intragastric transit and colonization of the epithelial gastric mucosa has never been analyzed in detail. We therefore infected gerbils, whose intragastric pH and response to infection resemble those of man, with H. pylori G1.1 wild type bacteria and their corresponding isogenic ureI mutants. Inhibitors of gastric acid secretion and colonization were used for manipulation of gastric pH. Gastric colonization was determined by urease assay and PCR. Gastric pH was measured with pH electrodes. Whereas H. pylori wild type or ureI complemented ureI knockout bacteria colonized the antrum, ureI deletion mutants were unable to colonize. However, continuous inhibition of acid secretion resulted in gastric colonization by the ureI mutants, as also observed with the wild type strain. Restoration of acid secretion resulted in eradication of ureI mutants but not wild type bacteria. The data show that ureI is essential for both gastric transit after inoculation and mucosal colonization in the untreated stomach. The eradication of ureI mutants following restoration of acid secretion suggests that the organism is exposed to pH < 4.0 at the surface of the antral mucosa and that UreI provides a target for specific monotherapy of H. pylori infections. PMID- 12558186 TI - Dependency of sugar transport and phosphorylation by the phosphoenolpyruvate dependent phosphotransferase system on membranous phosphatidyl glycerol in Escherichia coli: studies with a pgsA mutant lacking phosphatidyl glycerophosphate synthase. AB - It has been reported that phosphatidyl glycerol (PG) is specifically required for the in vitro activities of the hexose-phosphorylating Enzymes II of the Escherichia coli phosphoenolpyruvate-dependent sugar transporting phosphotransferase system (PTS). We have examined this possibility by measuring the properties of a null pgsA mutant that lacks detectable PG. The mutant showed lower in vitro phosphorylation activities towards several sugars when both PEP dependent and sugar-phosphate-dependent [14C]sugar phosphorylation reactions were measured. The order of dependency on PG for the different enzymes II was: IIMannose > IIGlucose > IIFructose > IIMannitol. Nonsedimentable (40000 rpm for 2 h) Enzymes II exhibited a greater dependency on PG than pelletable Enzymes II. Western blot analyses showed that the glucose Enzyme II is present in normal amounts. Transport and fermentation measurements revealed diminished activities for all Enzymes II. Thermal stability of all of these enzymes except the mannitol specific Enzyme II was significantly decreased by the pgsA mutation, and sensitivity to detergent treatments was enhanced. Sugar transport proved to be the most sensitive indicator of proper Enzyme II-phospholipid association. Our results show that PG stimulates but is not required for Enzyme II function in E. coli. PMID- 12558187 TI - Identification of Legionella species by ribotyping and other molecular methods. AB - There are currently more than 40 species of Legionella and the identification of most of them by standard methods is often technically difficult. The aim of this study was to use a ribotyping method with endonuclease HindIII and a probe consisting of a set of five oligonucleotides (referred to as OligoMix5). A total of 123 strains, including 78 type or reference strains corresponding to 44 species, eight clinical and 37 environmental isolates were tested. The usefulness of the method was demonstrated for the identification at the species level of all of the 123 Legionella isolates tested, with each species showing a specific profile. Among the 15 serogroups of Legionella pneumophila, eight patterns were obtained. For the 45 field strains, the randomly amplified polymorphic DNA (RAPD) technique and intergenic 16S-23S ribosomal spacer PCR analysis (ITS 16-23S) were also used. Altogether, these three methods allowed the identification of all of strains tested. However, ribotyping has proven to be more effective than the other methods. PMID- 12558188 TI - Identification of streptococcus mitis group species by RFLP of the PCR-amplified 16S-23S rDNA intergenic spacer. AB - Mitis group streptococci are pioneer colonizers of tooth surfaces and are implicated in various pathologies. Thus, accurate identification of oral mitis group strains would be valuable for studies of plaque ecology and dental caries and for diagnostic use in endocarditis or sepsis patients. The aim of this study was to evaluate the usefulness of PCR-RFLP analysis of the 16S-23S intergenic spacer for differentiating and identifying streptococcus mitis group species. The 16S-23S rDNA spacer regions of 27 type and reference Streptococcus strains, representing 8 species, were studied by PCR-mediated amplification by using oligonucleotide primers FGPS 1490-72 and FGPL 132'-38. PCR products were digested, independently, with 14 restriction enzymes. Only AluI, MboI, CfoI, HinfI and MaeII distinguished some species, particularly AluI and CfoI, but not all the species. Eight clusters were clearly generated, corresponding to currently recognized species, but only with the addition of five ITS restriction patterns, generated by AluI + MboI + CfoI + HinfI + MaeII, then clustered by UPGMA, on a distance consensus matrix. The combination of these five ITS RFLP tests allowed a relatively conclusive genomic group differentiation of mitis group species. Despite this observation, more strains of each species will need to be analyzed, particularly clinical isolates, before arriving at general conclusions about the utility of ITS restrictions for identification of strains at the species level. An ITS PCR-RFLP-based identifying method for streptococcus mitis group species would provide significant advantages over other molecular taxonomic methods which require DNA extraction and DNA-DNA hybridization. PMID- 12558189 TI - Nitric oxide synthase immunoreactivity of interstitial cells of Cajal in experimental colitis. AB - OBJECTIVE: There is functional and morphological evidence that interstitial cells of Cajal (ICC) may play a role in nitric oxide (NO) dependent signal transduction. However, little is known about the nitric oxide synthase (NOS) containing ICC during inflammation. MATERIALS AND METHODS: Immunocytochemical methods were used for the ultrastructural localization of NOS1-containing ICC in the wall of the colon of rats in experimental colitis. RESULTS: Large numbers of NOS immunoreactive (IR) nerve terminals were found in very close vicinity to smooth muscle cells as well as to blood vessels. IR nerves were found in close relationship with the ICC. The gap between the NOS IR nerve fibers and the membrane of smooth muscle cells and of ICC was 20-250 nm. In experimental colitis the number of NOS IR nerve fibers slightly decreased, however, large numbers (24%) of the ICC became IR for NOS. In the noninflamed area and in the controls, all these cells were immunonegative for NOS. CONCLUSIONS: Our light- and ultrastructural study suggests that some of the ICC can also synthesize NO, at least during inflammation. Therefore the change in the number and structure of ICC could play an important role in the pathogenesis of a variety of motility disorders. PMID- 12558190 TI - Spatial variation of plasma flow in the oxazolone-stimulated microcirculation. AB - INTRODUCTION: In cutaneous lymphocytic inflammation, enhanced regional blood flow is suggested by persistent erythema and warmth. Direct assessment of the microcirculation, however, has been limited by tissue edema and skin thickness. METHODS: To assess the microcirculatory adaptations to the epicutaneous antigen oxazolone, we studied the first pass kinetics and microvascular topography of the inflammatory skin microcirculation using a specially adapted epi-illumination intravital microscopy system. The fluorescence intravital videomicroscopy and streaming image acquisition of fluorescein-labeled dextran (approximately 500,000 MW) injections were used to assess changes in plasma flow. RESULTS: Direct plasma tracer injections of both the oxazolone-stimulated and control microcirculation demonstrated comparable transit times (leading edge and intensity-weighted peak times) from the carotid artery to the superficial vascular plexus (p > 0.05). In contrast to transit times, continuous infusion of the plasma tracer demonstrated a significant increase in the delivery of the fluorescein-labeled dextran to the oxazolone-stimulated microcirculation. Quantitative morphometry of intravital microscopic images demonstrated a 2.2-fold increase in the mean diameter of vessels in the superficial vascular plexus (p < 0.01). Further, fluorescence intensity mapping indicated that the increase was associated with increased perfusion of focal regions of the superficial vascular plexus (p < 0.001). CONCLUSIONS: These results indicate that the oxazolone-stimulated adaptations of the inflammatory microcirculation include both microvascular dilatation and the redistribution of plasma flow. PMID- 12558191 TI - Magnitude of the inflammatory response to cardiopulmonary bypass and its relation to adverse clinical outcomes. AB - INTRODUCTION: Cardiopulmonary bypass (CPB) induces an inflammatory response believed to contribute to postoperative morbidity. We hypothesized that the magnitude of the inflammatory response following CPB would be associated with adverse clinical outcomes. METHODS: Twenty-nine patients had plasma TNF, IL-6, IL 8, elastase, histamine, complement C5a, and complement C3a measured by ELISA before, during, and after cardiac operations employing CPB. Inflammatory mediator levels were analyzed with respect to outcomes. RESULTS: Mediator levels peaked at 4 h post-CPB and either returned to baseline or substantially decreased by 24 h. Patients with peak mediator levels above the median for the group as a whole were classified as 'hyper-responders'; those with levels below the median were classified as 'normal responders'. While IL-8, C3a, and IL-6 levels were independently associated with adverse outcomes, TNF, histamine, and C5a levels were not. Elastase levels trended towards adverse outcomes. IL-8 'hyper responders' experienced significantly greater postoperative weight gain and had higher IL-8 levels at 24 h (p<0.05), with trends towards renal impairment and protracted supplemental oxygen requirements. C3a 'hyper-responders' strongly trended towards increased bleeding, delayed extubation, greater postoperative weight gain, and decreased levels of independent functioning at discharge (p < or = 0.10). IL-6 'hyper-responders' experienced significantly more postoperative bleeding, delayed extubation, and higher IL-6 levels at 24 h compared to 'normal responders' (p < 0.05). They strongly trended towards greater postoperative weight gain and decreased levels of independent functioning at discharge (p < or = 0.10). CONCLUSIONS: Patients who have an exaggerated inflammatory response to CPB tend to bleed more, require more respiratory support, demonstrate greater capillary leak via weight gain, and display a decline in independent functioning relative to normal responders. Thus, it appears that the magnitude of the inflammatory response to CPB adversely influences clinical outcomes. PMID- 12558192 TI - Characteristics of histamine tachyphylaxis in rat uterine smooth muscle. AB - OBJECTIVE AND DESIGN: To study both the desensitisation induced by short-term exposure to histamine and the mechanism responsible in the isolated rat uterus. MATERIAL: Precontracted isolated uterus (37 mM KCl) from oestrogenised Wistar rats were used. TREATMENT: Repetitive responses to histamine (10(-6), 10(-5), 10( 4), 10(-3) M), dimaprit and clonidine (10(-4) M) were tested at 15, 30, 45 and 105 min., with their modifications by (5 mg/kg, 24 h before sacrifice) reserpine, 10(-7) M propranolol, 10(-8) M atropine, and 10(-6) M indomethacin. Dose-response curves for adrenaline were carried out as standard protocol. METHODS: In vitro techniques (de Jalon's solution, 31 degrees C, carbogen, isotonic registration, resting tension 1 g). Levels of cAMP were studied in response to histamine, adrenaline and isoproterenol by radioassay. RESULTS: Repeated histamine produced a rapid loss of inhibitory uterine response depending on the number of exposures and increase in concentration. The tachyphylaxis phenomenon is not modified by the different pretreatments used. Dimaprit, but not clonidine or adrenaline, produced tachyphylaxis. The second exposition to 10(-3) M histamine produced a 38.8% reduction in cAMP production. CONCLUSION: Histamine appears to induce homologous and probably cAMP-dependent desensitisation of H2-receptors. PMID- 12558193 TI - Endogenous adenosine regulates neutrophil pro-inflammatory activities by cyclic AMP-dependent accelerated clearance of cytosolic calcium. AB - OBJECTIVE AND DESIGN: To identify the involvement of adenosine in restoration of Ca2+ homeostasis to activated human neutrophils. MATERIALS: Neutrophils were isolated from venous blood taken from healthy, adult, human volunteers. TREATMENT: The cells were exposed to adenosine deaminase (ADA, 0.1-2 units/ml) for 10 min at 37 degrees C prior to activation with N-formyl-L-methionyl-L-leucyl L-phenylala-nine (FMLP, 1 microM). METHODS: Cytosolic Ca2+ concentrations and transmembrane fluxes of the cation in FMLP-activated neutrophils +/- ADA were measured using spectrofluorimetric and radiometric procedures respectively, while intracellular cAMP and inositol triphosphate were measured by radioassay, and superoxide production and elastase release by, chemiluminescence and colourimetric methods respectively. Levels of statistical significance were calculated using the Mann-Whitney U-test and ANOVA. RESULTS: Although FMLP activated generation of inositol triphosphate and mobilisation of Ca2+ from neutrophil internal stores, as well as the magnitude of the subsequent efflux and store-operated influx of the cation were unaffected by ADA, there was a prolonged elevation in cytosolic Ca2+ in the presence of the enzyme, which was associated with failure to activate adenylate cyclase and with increased production of superoxide and release of elastase. These effects of ADA were attenuated by dibutyryl cAMP (4 mM), CGS 21680 (1 microM) and rolipram (0.5 microM), as well as by EGTA (10 mM). CONCLUSIONS: These results are compatible with a physiological role for adenosine in promoting deactivation of neutrophils, possibly by promoting cAMP-dependent clearance of Ca2+ from the cytosol of the cells by the endo-membrane Ca2+-ATPase. PMID- 12558195 TI - Gene expression profiling of Ca2+-atpase inhibitor DTBHQ and antigen-stimulated RBL-2H3 mast cells. AB - OBJECTIVE AND DESIGN: Ca2+ signaling is critical for mast cell activation by antigen stimulation, and we previously described that the signaling can be mimicked by Ca2+-ATPase inhibitors. We therefore investigated the effect of the Ca2+-ATPase inhibitor and antigen stimulation on the gene expression profiles of RBL-2H3 mast cells. MATERIAL: A Ca2+-ATPase inhibitor, 2,5-di(tert-butyl)-1,4 hydroquinone (DTBHQ), an antigen (dinitrophenylated BSA), a high-density oligonucleotide microarray (Affymetrix GeneChip) technique, and a well characterized rat mast cell line RBL-2H3 were used. TREATMENT: RBL-2H3 cells were activated for 3 h with 10 microM DTBHQ, which increases cytosolic Ca2+ concentration, or 10 microg/ml antigen, which cross-links IgE receptors, and the mRNA expression profiles (8,799 genes) were analyzed with GeneChip arrays (n = 3). METHODS: Expression levels were measured by GeneChip, and the differences were tested by Welch's t-test and P-values less than 0.05 were considered statistically significant. Values are expressed as means +/- SEM. RESULTS: The genes, including MCP-1, GADD45, Relaxin H1, CSF-1, c-jun-oncogene, Pyk-2, NKR-P2 and CREM, were significantly up-regulated by both DTBHQ and antigen stimuli, whereas the genes including interleukin (IL)-3, IL-4, IL-9, IL-13, GADD153, butyrate response factor, and Fas ligand, were up-regulated by DTBHQ alone. On the other hand, the expression of several genes, including GATA-1, were down regulated by DTBHQ stimulation. CONCLUSIONS: These results suggest 1) that DTBHQ seems to induce proinflammatory responses by stimulating the production of several cytokines through the expression of several transcription factors, 2) that the changes in gene expression profile induced by DTBHQ and by IgE receptor cross-linking in mast cells were almost the same, but many more stress-inducible genes like GADD 153 were up-regulated by the former. PMID- 12558194 TI - Effect of the selective COX-2 inhibitors, celecoxib and rofecoxib in rat acute models of inflammation. AB - OBJECTIVE: This study evaluates the action of celecoxib and rofecoxib, two selective cyclooxygenase-2 (COX-2) inhibitors in two acute models of inflammation, carrageenan (Cg)-induced rat pleurisy, and paw oedema formation. MATERIAL: Male Wistar rats (N = 4-10 per group) were used. A fixed volume of PBS or carrageenan was injected into the pleural cavity or into the paw. Furthermore, the myeloperoxidase (MPO) activity and the levels of nitrite/nitrate (NOx), interleukin-1beta (IL-1beta), tumor necrosis factor-a (TNF-a) and PGE2 were also assessed in the paw tissue or in pleural exudate. RESULTS: Dexamethasone (DEX, 0.5 mg kg(-1), s.c., -4 h) and indomethacin (INDO, 3 mg kg(-1), p.o., -1 h) suppressed Cg-induced pleural exudate accumulation by 84 and 77% and inflammatory cell influx by 66 and 47%, respectively. In contrast, celecoxib (CLX, 10 mg kg( 1), p.o., -1 h) or rofecoxib (RFX, 10 mg kg(-1) , p.o., -1 h) only reduced the Cg induced pleural exudate volume by 44 and 40%, respectively, but had no significant effect over inflammatory cell influx. At the same doses used for pleurisy, DEX, INDO, CLX, RFX and SC-560 (a selective COX-1 inhibitor, 40 mg kg( 1), p.o., -1 h), inhibited the Cg-induced paw oedema by 49, 31, 21, 21 and 17%. DEX, INDO or SC-560 reduced the level of MPO by 71, 78 and 59%, while CLX or RFX produced a small, but significant increase (28 or 16%) in MPO activity. In the rat model of pleurisy, PGE2 levels in cell-free exudates were significantly attenuated by 91, 89, 57 and 65% in animals treated with DEX, INDO, CLX or RFX. In contrast, INDO reduced significantly the whole bloodTXB, synthesis (59%) while DEX and INDO reduced the pleural content of NOx significantly. Treatment of animals with CLX or RFX did not alter the content of pro-inflammatory cytokines IL-1beta or TNF-alpha in the pleural exudate, but CLX reduced IL-1beta levels in the rat paw tissue and RFX increased TNF-alpha in this tissue. CONCLUSION: Together these results provide consistent evidence indicating that the selective COX-2 inhibitors CLX and RFX, in contrast to DEX, INDO or SC-560, despite reducing greatly the Cg-induced pleural exudation, PGE2 content and paw oedema have only partial acute anti-inflammatory properties in two different rat acute models of inflammation. PMID- 12558196 TI - Individual differences in self-assessed health: an information-processing investigation of health and illness cognition. AB - In 2 studies, the relation between measures of self-assessed health (SAH) and automatic processing of health-relevant information was investigated. In Study 1, 84 male and 86 female undergraduate students completed a modified Stroop task. Results indicated that participants with poorer SAH showed enhanced interference effects for illness versus non-illness words. In Study 2, 27 male and 30 female undergraduate students completed a self-referent encoding task. Results offered a conceptual replication and extension of Study 1 by confirming the specificity of the relation between SAH measures and automatic processing of health (vs. negative or positive general trait) information. These studies provide evidence that individual differences in SAH are reflected in schematic processing of health-relevant information. PMID- 12558197 TI - Predictors of repeat hospitalizations in children with asthma: the role of psychosocial and socioenvironmental factors. AB - This study examined relationships between psychosocial factors and asthma rehospitalization patterns in 115 children (ages 4-15) who had > or = 1 hospitalization during the study period. Lifetime history of hospitalizations and new hospitalizations during a 1-year follow-up period were measured, controlling for baseline asthma symptoms and medications. Prospectively, caretaker characteristics (lower sense of mastery, being less emotionally bothered by asthma) predicted greater likelihood of future asthma hospitalizations. Lifetime history of hospitalizations was associated with family impacts (greater family strain and family conflict greater financial strain) as well as caretaker characteristics (greater personal strain, beliefs about not being able to manage one's child's asthma). These findings could help guide future interventions targeted at the subgroup of children who represent a high proportion of asthma hospitalizations. PMID- 12558198 TI - Occupation and subclinical carotid artery disease in women: are clerical workers at greater risk? AB - The current study examined cardiovascular risk factors and carotid atherosclerosis in 362 women (ages 42-50 years) who were working in clerical, blue-collar, or white-collar jobs or who were not employed. Risk factors were measured premenopausally and ultrasound measures of carotid atherosclerosis were obtained approximately 11 years later. Clerical and blue-collar workers had more atherogenic profiles on physical, behavioral, and psychosocial risk factors when compared with white-collar and nonemployed women. Clerical workers had greater carotid intima-media thickness relative to all other groups and more focal plaque when compared with white-collar workers. Risk factors and workplace characteristics did not account for the greater carotid atherosclerosis observed in clerical workers. Further research is needed to investigate why clerical work may increase cardiovascular risk. PMID- 12558199 TI - Cognitive adaptation, psychological adjustment, and disease progression among angioplasty patients: 4 years later. AB - The long-term effects of the Cognitive Adaptation Theory Index (CATI) on psychological and physical health outcomes among men (n = 199) and women (n = 99) treated for coronary artery disease with percutaneous transluminal coronary angioplasty were examined. The CATI reflects a positive view of the self, a positive view of one's future, and a sense of personal control over daily life. This index was created from questionnaires administered during hospitalization for the initial angioplasty. Four years later, the CATI predicted positive adjustment to disease, even when initial adjustment was taken into consideration. In addition, the CATI predicted a reduced likelihood of sustaining a subsequent cardiac event over 4 years. This association was more robust for men. PMID- 12558200 TI - Body image and strategies to lose weight and increase muscle among boys and girls. AB - This study examined factors that influence body image and strategies to either lose weight or increase muscle among children. Participants were 237 boys and 270 girls. Body mass index (BMI), body dissatisfaction, cognitions and behaviors to both lose weight and increase muscles, as well as self-esteem and positive and negative affect, were evaluated. Self-esteem was associated with body satisfaction, positive affect predicted strategies to lose weight and increase muscles, and negative affect predicted body dissatisfaction and cognitions to lose weight and increase muscles. Boys were more likely to focus on changing muscles. Respondents with higher BMIs were more focused on losing weight butnot muscle. The discussion focuses on health risk behaviors related to eating and exercise among children. PMID- 12558201 TI - Are changes in blood pressure and total cholesterol related to changes in mood? An 18-month study of men and women. AB - The authors investigated the within-person association of reported mood with blood pressure and total cholesterol (TC) levels, each assessed 4 times over an 18-month period in 128 men and 154 women. Change over time in tense arousal was significantly positively associated with changes over time in systolic blood pressure (SBP) and diastolic blood pressure (DBP) but not TC. A change in hedonic tone was significantly associated with SBP (an increase in negative affect was associated with an increase in SBP) but not with DBP or TC. There were no sex differences in associations of mood with SBP or TC. However, increases in tense arousal and negative affect were significantly associated with an increase in DBP for women but not men. PMID- 12558202 TI - Psychological impact of colorectal cancer screening. AB - This article examines the psychological impact of participating in sigmoidoscopy screening for colorectal cancer prevention. The 1st study examined psychological well-being at 3 months, in relation to screening outcome, in 4,153 individuals. The 2nd study used longitudinal data to examine changes in psychological functioning from before to after screening in relation both to screening outcome and baseline indicators of vulnerability. There were few psychological differences between those who had received negative results or had polyps detected. These findings were confirmed in the longitudinal study, which also found no evidence for vulnerability to adverse effects among those who were initially most anxious or who perceived their risk of cancer to be higher. The longitudinal data suggested that screening might produce transient positive effects. PMID- 12558203 TI - Using message framing to motivate HIV testing among low-income, ethnic minority women. AB - This study compared the effectiveness of 4 videotaped educational programs designed to motivate HIV testing among low-income, ethnic minority women. Four hundred eighty women were assigned randomly to watch one of 2 gain-framed or 2 loss-framed videos. Consistent with prospect theory, participants' perceptions of the certainty of the outcome of an HIV test moderated the effects of framing on self-reported testing behavior 6 months after video exposure. Among participants who reported being certain of the test's outcome, those who saw a gain-framed video reported a higher rate of testing than those who saw a loss-framed message. Among women who perceived the outcome of HIV testing as relatively uncertain, gain- and loss-framed videos led to similar rates of self-reported testing, with some advantage for the loss-framed message. PMID- 12558204 TI - Demonstrations of implicit anti-fat bias: the impact of providing causal information and evoking empathy. AB - Three studies investigated implicit biases, and their modifiability, against overweight persons. In Study 1 (N = 144), the authors demonstrated strong implicit anti-fat attitudes and stereotypes using the Implicit Association Test, despite no explicit anti-fat bias. When participants were informed that obesity is caused predominantly by overeating and lack of exercise, higher implicit bias relative to controls was produced; informing participants that obesity is mainly due to genetic factors did not result in lower bias. In Studies 2A (N = 90) and 2B (N = 63), participants read stories of discrimination against obese persons to evoke empathy. This did not lead to lower bias compared with controls but did produce diminished implicit bias among overweight participants, suggesting an in group bias. PMID- 12558205 TI - Patterns and correlates of binge drinking trajectories from early adolescence to young adulthood. AB - Latent growth mixture modeling was used to identify developmental trajectories (described in terms of demographics, exposure and resistance to a pro-drug environment, and deviant behavior) of binge drinking among 5,694 individuals who completed 6 surveys from ages 13 to 23 years: nonbingers (32%); moderate stables (37%), who had consistently low levels of bingeing; steady increasers (16%), who increased from the lowest to highest level of bingeing; adolescent bingers (9%), whose early rise in bingeing was followed by a decrease to a moderate level; and early highs (6%), who decreased from the highest level of bingeing to a moderate level. Results show considerable diversity in binge drinking patterns and the correlates of bingeing across trajectory classes. PMID- 12558206 TI - Correlates of unhealthy weight-control behaviors among adolescents: implications for prevention programs. AB - This study aimed to identify correlates of unhealthy weight-control behaviors in adolescents to guide the development of programs aimed at the primary prevention of disordered eating. A model explaining unhealthy weight-control behaviors was tested among 4,746 adolescents using structural equation modeling. Models fit the data well and explained 76% of the variance in unhealthy weight-control behaviors among girls and 63% among boys. Weight-body concerns were a strong correlate of unhealthy weight-control behaviors in both girls and boys. Models also emphasized the importance of weight-specific social norms within the adolescent's proximal environment. Findings suggest the importance of addressing weight-body concerns within prevention programs and extending interventions beyond classroom settings to ensure changes in weight-related norms among peer groups and family members. PMID- 12558207 TI - Increasing attendance at colorectal cancer screening: testing the efficacy of a mailed, psychoeducational intervention in a community sample of older adults. AB - This article describes a trial of a psychoeducational intervention designed to modify negative attitudes toward flexible sigmoidoscopy screening and thereby increase screening attendance. The intervention materials addressed the multiple barriers shown to be associated with participation in earlier studies. Adults ages 55-64 (N = 2,966), in a "harder-to-reach" group were randomized either to receive an intervention brochure or to a standard invitation group. Attitudes and expectations were assessed by questionnaire, and attendance at the clinic was recorded. Compared with controls, the intervention group had less negative attitudes, anticipated a more positive experience, and had a 3.6% higher level of attendance. These results indicate that psychoeducational interventions can provide an effective means of modifying attitudes and increasing rates of screening attendance. PMID- 12558208 TI - Development of the cognitive appraisal and understanding of social events (CAUSE) videos. AB - This study describes the development and validation of 4 videos designed to assess adolescent cognitive appraisals and understanding of social events (the CAUSE Videos). Story lines varied in outcome (2 negative and 2 ambiguous). Convergent and divergent validity were tested in samples of college freshmen and sophomores. As hypothesized, threatening interpretations during ambiguous situations were positively associated with trait hostility (convergent validity) but were not significantly associated with aggression (divergent validity). Negative video interpretations were not significantly associated with hostility or aggression. These videos provide a valid method for assessing adolescents' appraisals of ambiguous and negative social situations in a laboratory setting and may be used to test hypotheses about cognitive processes underlying associations such as that between low socioeconomic status and health in adolescents. PMID- 12558209 TI - Health-related Internet use, coping, social support, and health indicators in people living with HIV/AIDS: preliminary results from a community survey. AB - Widespread Internet use has revolutionized health information and patient education for persons with chronic illnesses. The authors surveyed 147 HIV positive persons to examine factors associated with Internet use and associations between Internet use and health. Information, motivation, and behavioral skills associated with using the Internet were related to Internet use. The authors found that health-related Internet use was associated with HIV disease knowledge, active coping, information seeking coping, and social support among persons who were using the Internet. These preliminary findings suggest an association between using the Internet for health-related information and health benefits among people living with HIV/AIDS, supporting the development of interventions to close the digital divide in HIV/AIDS care. PMID- 12558210 TI - Selection fairness information and applicant reactions: a longitudinal field study. AB - Although laboratory studies have found that selection information can affect applicant perceptions, this has not been tested in the field. The authors followed 2 cohorts of police applicants (N = 274) in a longitudinal study to examine the relationship between information, applicant perceptions, and behavior (e.g., turnover). Information was related to perceived fairness measured at the time of testing and 1 month later when applicants received their results. Information moderated the relationship between outcome favorability and test taking self-efficacy among African Americans but not among Whites. Information was not related to the behavioral measures. The discussion focuses on why certain findings from previous studies were not replicated and on the use of information when applicants have an investment in getting a job. PMID- 12558211 TI - Reducing the performance-cue bias in work behavior ratings: can groups help? AB - The authors examined whether the performance-cue bias can be reduced by relying on groups as raters. Study participants (N = 333) were provided with feedback regarding the performance of a workgroup and, after observing the group, assigned to an individual or group rater condition to complete a behavioral rating instrument. Results revealed that when provided with positive (vs. negative) feedback, individuals attributed more effective and fewer ineffective behaviors to the workgroup; however, group ratings were unaffected by the feedback. In addition, feedback biased the decision criteria and false alarm rates of individuals but not of groups. Discussion of when groups may attenuate versus amplify bias in performance appraisal judgments emphasizes 2 key elements-bias magnitude and task perception. PMID- 12558212 TI - How effective are the cross-examination and expert testimony safeguards? Jurors' perceptions of the suggestiveness and fairness of biased lineup procedures. AB - Mock jurors (N = 800) viewed a videotaped trial that included information about a lineup identification procedure. Suggestiveness of the eyewitness identification procedure varied in terms of foil, instruction, and presentation biases. Expert testimony regarding the factors that influence lineup suggestiveness was also manipulated. Criteria included juror ratings of lineup suggestiveness and fairness, ratings of defendant culpability, and verdicts. Jurors were sensitive to foil bias but only minimally sensitive to instruction and presentation biases. Expert testimony enhanced juror sensitivity only to instruction bias. These results have implications for the effectiveness of cross-examination and expert testimony as safeguards against erroneous convictions resulting from mistaken identifications. PMID- 12558214 TI - Abusive supervision and subordinates' organizational citizenship behavior. AB - The relationship between subordinates' perceptions of abusive supervision and supervisors' evaluations of subordinates' organizational citizenship behavior (OCB) was explored among a sample of 373 Air National Guard members and their military supervisors. As predicted, the relationship between abusive supervision and subordinates' OCB was stronger among subordinates who defined OCB as extra role behavior (compared with those defining OCB as in-role behavior), and this effect was fully mediated by the interactive effect of procedural justice and OCB role definitions. The study's implications for theory and research are discussed, its limitations are identified, and directions for future research are suggested. PMID- 12558213 TI - The influence of positive affect on the components of expectancy motivation. AB - The influence of positive affect on expectancy motivation was investigated in 2 studies. The results of Study 1 indicated that positive affect improved people's performance and affected their perceptions of expectancy and valence. In Study 1, in which outcomes depended on chance, positive affect did not influence people's perceptions of instrumentality. In Study 2, in which the link between performance and outcomes was specified, positive affect influenced all 3 components of expectancy motivation. Together, the results of Studies 1 and 2 indicated that positive affect interacts with task conditions in influencing motivation and that its influence on motivation occurs not through general effects, such as response bias or general activation, but rather through its influence on the cognitive processes involved in motivation. PMID- 12558215 TI - Context and cue cards in young children's testimony: a comparison of brief narrative elaboration and context reinstatement. AB - In a comparison of interview procedures that aimed to assist retrieval strategies in children's event recall, 72 children aged 4-6 years were questioned about a witnessed event. Context reinstatement and brief narrative elaboration (an abbreviated version of K. J. Saywitz and L. Snyder's [1996] procedure) produced similar levels of correct recall, and both elicited more correct recall than did a control condition, with no increase in errors. Combining these procedures did not further improve performance. The superiority of narrative elaboration over the control group was evident in free recall and did not depend on explicit prompting with cue cards. These findings suggest that incorporating brief narrative elaboration training in investigative interviews with children may, like context reinstatement, be a valuable mnemonic aid. PMID- 12558216 TI - Identity salience and the influence of differential activation of the social self schema on advertising response. AB - The authors examined how identity primes and social distinctiveness influence identity salience (i.e., the activation of a social identity within an individual's social self-schema) and subsequent responses to targeted advertising. Across 2 studies, individuals who were exposed to an identity prime (an ad element that directs attention to the individual's social identity) and who were socially distinctive (minorities in the immediate social context) expressed systematically different evaluations of spokespersons and the advertisements that featured them. Specifically, Asian (Caucasian) participants responded most positively (negatively) to Asian spokespeople and Asian-targeted advertising when the participants were both primed and socially distinctive. No main effects of identity primes or social distinctiveness were found. The implications of these findings for identity theory, advertising practice, and intervention communications are discussed. PMID- 12558217 TI - Predictive validity of a multidisciplinary model of reemployment success. AB - The authors propose a multidisciplinary model of the predictors of reemployment and test its predictive validity for explaining reemployment success. Predictor variables from the fields of economics, sociology, and psychology are incorporated into the model. Reemployment success is conceptualized as a construct consisting of unemployment insurance exhaustion and reemployment speed, and for reemployed persons, job improvement, job-organization fit, and intention to leave the new job. Direct, mediated, and moderated relationships were hypothesized and tested, clarifying the role of the variables in the reemployment process and outcome. The authors' proposal and examination of a multidisciplinary model of reemployment success contributes to a literature that has not tended to adequately cross disciplinary boundaries. PMID- 12558218 TI - The relationship between early recruitment-related activities and the application decisions of new labor-market entrants: a brand equity approach to recruitment. AB - Theory and research from the marketing literature on customer-based brand equity were used to predict how positive exposure to 4 early recruitment-related activities-publicity, sponsorships, word-of-mouth endorsements, and advertising may affect the application decisions of engineering students. Similar to prior marketing findings, the results suggested that early recruitment-related activities were indirectly related to intentions and decisions through 2 dimensions of employer brand image: general attitudes toward the company and perceived job attributes. The relationships between word-of-mouth endorsements and the 2 dimensions of brand image were particularly strong. In addition, it was found that early recruitment-related activities interacted with one another such that employer brand image was stronger when firms used publicity in conjunction with other early recruitment-related activities. PMID- 12558219 TI - A hierarchical taxonomy of top managers' goals. AB - To embed goal theories more deeply in the domain of top-level leadership behavior and to provide a vehicle to facilitate future research, the authors developed a taxonomy of managerial goals. Interviews with 75 company leaders-founders and presidents-from 3 countries generated 2,182 articulated goals. Content analysis supported 2 taxonomic dimensions: goal content and hierarchical level. The goal content dimension specified 10 categories of substantive goal targets, and the second dimension captured the hierarchical structure of the top leaders' goal sets, with lower-level goals being instrumental toward achieving superordinate goals. The hierarchy comprised 5 goal levels: ultimate, enterprise, strategic, project, and process. Chi-square analyses revealed relationships between goal content and hierarchical level as well as differences between the national subsamples. PMID- 12558220 TI - Job-decision inconsistencies involving social comparison information: the role of dominating alternatives. AB - Previous research on joint versus separate preference reversals has demonstrated that individuals focus on social comparison information when they are evaluating a single option but focus on absolute salary when they are considering more than 1 option. Study 1 demonstrates that social comparison information is important in multiple option scenarios when an option favorable on social comparison dominates an inferior, 3rd alternative. Study 2 examines why dominating alternatives are so attractive by investigating the role that the value-shift, weight-change, and emergent-value models play in explaining the pattern of results obtained in Study 1. Results provide support for the value-shift and emergent-value models and further suggest that these 2 models may be interrelated, with justification (emergent-value model) mediating the relationship between the attractiveness of the attributes (value-shift model) and the attractiveness of the dominating alternative. PMID- 12558221 TI - Drug testing, drug treatment, and marijuana use: a fairness perspective. AB - The authors conducted a random statewide telephone survey of 1,484 individuals to study the relationship between marijuana use (in terms of participants' history of marijuana use) and reactions to drug testing and to study 2 hypothetical drug treatment policies. Job safety sensitivity was related to perceived fairness of drug testing for the participant's job, and more recent marijuana use was associated with more negative reactions. Safety sensitivity was related to perceived fairness of drug treatment. Organizations with voluntary treatment were more attractive than ones with monitored treatment. Marijuana use interacted with drug treatment policy type in predicting reactions to drug treatment. Results suggest that organizations should consider job and employee characteristics when developing a drug treatment policy. PMID- 12558222 TI - The role of employee loyalty and formality in voicing discontent. AB - This study extends prior research on voice, loyalty, and postvoice outcomes by examining a variety of methods by which an employee may choose to voice a dispute. The authors argue that more loyal employees may prefer and use less formal methods to voice discontent and that the use of less formal voice methods relates to less job search activity and lower intent to quit. This study also investigated the interrelation between loyalty, voice method, and satisfaction with the attempt to resolve the dispute. Findings from a sample of university staff employees (N = 452) provide support for the hypotheses. PMID- 12558223 TI - Relationships between conscientiousness, self-efficacy, self-deception, and learning over time. AB - The present study examined the dual mediating effects of self-efficacy and self deception on the relationship between conscientiousness and learning over time. Data from 134 college students were used to investigate the relative impact of self-efficacy and self-deception. Consistent with the hypothesized model, conscientiousness was significantly and positively related to both early training self-efficacy and self-deception, and both self-efficacy and self-deception had significant effects on learning but in opposite directions. Furthermore, the relative impact of self-efficacy and self-deception on learning changed over time as expected. The negative effect of self-deception in early stages of training disappeared at later stages of training but the positive effects of self-efficacy remained. Support was not found for self-efficacy and self-deception as mediators of the conscientiousness-learning relationship. PMID- 12558224 TI - The moderating effects of employee tenure on the relation between organizational commitment and job performance: a meta-analysis. AB - This meta-analysis investigated the correlation between attitudinal commitment and job performance for 3,630 employees obtained from 27 independent studies across various levels of employee tenure. Controlling for employee age and other nuisance variables, the authors found that tenure had a very strong nonlinear moderating effect on the commitment-performance correlation, with correlations tending to decrease exponentially with increasing tenure. These findings do not appear to be the result of differences across studies in terms of the type of performance measure (supervisory vs. self), type of tenure (job vs. organizational), or commitment measure (Organizational Commitment Questionnaire [L. W. Porter, R. M. Steers, R. T. Mowday, & P. V. Boulian, 19741 vs. other). The implications and future research directions of these results are discussed. PMID- 12558225 TI - The disclosure dilemma for gay men and lesbians: "coming out" at work. AB - This study examined disclosing sexual orientation at work for 220 gay men and 159 lesbians. Self-acceptance, the centrality of one's identity, how "out" one is to friends and family, employer policies, and perceived employer gay-supportiveness were associated with disclosure behaviors at work for gay/lesbian employees. Disclosing at work and working for an organization perceived to be more gay supportive was related to higher job satisfaction and lower job anxiety. Reactions of coworkers to gay or lesbian workers mediated the relationship between disclosure and gay/lesbian workers' job attitudes. Implications and solutions for management are discussed. PMID- 12558226 TI - The use of impression management tactics in structured interviews: a function of question type? AB - This study investigated impression management tactic use during structured interviews containing both experience-based and situational questions. Specifically, the authors examined whether applicants' use of impression management tactics depended on question type. Results from 119 structured interviews indicated that almost all of the applicants used some form of impression management. Significantly more assertive than defensive impression management tactics were used, and among assertive tactics, applicants tended to use self-promotion rather than ingratiation. However, different question types prompted the use of different impression management tactics. Ingratiation tactics were used significantly more when applicants answered situational questions, whereas self-promotion tactics were used significantly more when applicants answered experience-based questions. Furthermore, the use of self-promotion and ingratiation tactics was positively related to interviewer evaluations. PMID- 12558227 TI - Staphylococcus aureus bacteremia associated with intra-aortic balloon pumps. PMID- 12558228 TI - Exactly 250 feet. PMID- 12558229 TI - Surveillance, reporting, automation, and interventional epidemiology. PMID- 12558230 TI - Improvement of intraoperative antibiotic prophylaxis in prolonged cardiac surgery by automated alerts in the operating room. AB - OBJECTIVE: To assess the impact of an automated intraoperative alert to redose prophylactic antibiotics in prolonged cardiac operations. DESIGN: Randomized, controlled, evaluator-blinded trial. SETTING: University-affiliated hospital. PATIENTS: Patients undergoing cardiac surgery that lasted more than 4 hours after the preoperative administration of cefazolin, unless they were receiving therapeutic antibiotics at the time of surgery. INTERVENTION: Randomization to an audible and visual reminder on the operating room computer console at 225 minutes after the administration of preoperative antibiotics (reminder group, n = 137) or control (n = 136). After another 30 minutes, the circulating nurse was required to indicate whether a follow-up dose of antibiotics had been administered. RESULTS: Intraoperative redosing was significantly more frequent in the reminder group (93 of 137; 68%) than in the control group (55 of 136; 40%) (adjusted odds ratio, 3.31; 95% confidence interval, 1.97 to 5.56; P < .0001). The impact of the reminder was even greater when compared with the 6 months preceding the study period (129 of 480; 27%; P < .001), suggesting some spillover effect on the control group. Redosing was formally declined for 19 of the 44 patients in the reminder group without redosing. The rate of surgical-site infection in the reminder group (5 of 137; 4%) was similar to that in the control group (8 of 136; 6%; P = .42), but significantly lower than that in the pre-study period (48 of 480; 10%; P = .02). CONCLUSION: The use of an automatic reminder system in the operating room improved compliance with guidelines on perioperative antibiotic prophylaxis. PMID- 12558231 TI - A risk index for sternal surgical wound infection after cardiovascular surgery. AB - OBJECTIVES: To identify factors that increase the risk of sternal surgical wound infection after cardiovascular surgery and to develop a bedside clinical risk index using these factors. DESIGN: A risk index was developed using clinical data collected from a cohort of 11,508 cardiac surgery patients and validated using three independent subsets of the data. With two of these subsets, we derived a logistic regression equation and then modified the scoring algorithm to simplify the calculation of patient risk scores by clinicians. The final subset was used to validate the index. The area under the receiver operating characteristic (aROC) curve was the primary measure of goodness of fit. SETTING: Toronto General Hospital, a teaching hospital and the largest center for cardiac surgery in Ontario, Canada. PATIENTS: Cardiac surgery patients receiving cardiopulmonary bypass between April 1, 1990, and December 31, 1995, who survived at least 6 days after surgery. RESULTS: Variables that were used to construct the risk index included reoperation due to complication (odds ratio, 4.3; range, 1.9 to 8.5), diabetes (odds ratio, 2.4; range, 1.5 to 3.7), more than 3 days in the intensive care unit (odds ratio, 5.4; range, 3.2 to 8.7), and use of the internal mammary artery for revascularization (odds ratio, 3.2; range, 1.7 to 5.8). Validation showed that the index had an aROC curve of 0.64. CONCLUSIONS: The risk index described in this article allows clinicians to quickly stratify patients into four risk groups associated with an increasing risk of sternal surgical wound infection. It may be used perioperatively or as part of a wound infection surveillance system. PMID- 12558232 TI - Incidence of surgical-site infections and the validity of the National Nosocomial Infections Surveillance System risk index in a general surgical ward in Santa Cruz, Bolivia. AB - OBJECTIVES: To estimate the frequency of and risk factors for surgical-site infections (SSIs) in Bolivia, and to study the performance of the National Nosocomial Infections Surveillance (NNIS) System risk index in a developing country. DESIGN: A prospective study with patient follow-up until the 30th postoperative day. SETTING: A general surgical ward of a public hospital in Santa Cruz, Bolivia. PATIENTS: Patients admitted to the ward between July 1998 and June 1999 on whom surgical procedures were performed. RESULTS: Follow-up was complete for 91.5% of 376 surgical procedures. The overall SSI rate was 12%. Thirty-four (75.6%) of the 45 SSIs were culture positive. A logistic regression model retained an American Society of Anesthesiologists score of more than 1 (odds ratio [OR], 1.87), a not-clean wound class (OR, 2.28), a procedure duration of more than 1 hour (OR, 1.81), and drain (OR, 1.98) as independent risk factors for SSI. There was no significant association between the NNIS System risk index and SSI rates. However, a "local" risk index constructed with the above cutoff points showed a linear trend with SSI (P < .001) and a relative risk of 3.18 for risk class 3 versus a class of less than 3. CONCLUSIONS: SSIs cause considerable morbidity in Santa Cruz. Appropriate nosocomial infection surveillance and control should be introduced. The NNIS System risk index did not discriminate between patients at low and high risk for SSI in this hospital setting, but a risk score based on local cutoff points performed substantially better. PMID- 12558233 TI - Risk factors for spinal surgical-site infections in a community hospital: a case control study. AB - OBJECTIVE: To characterize risk factors for surgical-site infection after spinal surgery. DESIGN: A case-control study. SETTING: A 113-bed community hospital. METHOD: From January 1998 through June 2000, the incidence of surgical-site infection in patients undergoing laminectomy, spinal fusion surgery, or both increased at community hospital A. We compared 13 patients who acquired surgical site infections after laminectomy, spinal fusion surgery, or both with 47 patients who were operated on during the same time period but did not acquire a surgical-site infection. Information collected included demographics, risk factors, personnel involved in the operations, length of hospital stay, and hospital costs. RESULTS: Of 13 case-patients, 9 (69%) were obese, 9 (69%) had spinal compression, 5 (38.5%) had a history of tobacco use, and 4 (31%) had diabetes. Oxacillin-sensitive Staphylococcus aureus (6 of 13; 46%) was the most common organism isolated. Significant risk factors for postoperative spinal surgical-site infection were dural tear during the surgical procedure and the use of glue to cement the dural patch (3 of 13 [23%] vs 1 of 47 [2.1%]; P = .02) and American Society of Anesthesiologists risk class of 3 or more (6 of 13 [46.2%] vs 7 of 47 [15%]; P = .02). Case-patients were more likely to have prolonged length of stay (median, 16 vs 4 days; P< .001). The average excess length of stay was 11 days and the excess cost per case was $12,477. CONCLUSION: Dural tear and the use of glue should be evaluated as potential risk factors for spinal surgical-site infection. Systematic observation for potential lapses in sterile technique and surgical processes that may increase the risk of infection may help prevent spinal surgical-site infection. PMID- 12558234 TI - A prospective study of tobacco smoking as a predictor of complications in general surgery. AB - OBJECTIVE: To analyze whether tobacco smoking is related to nosocomial infection, admission to the intensive care unit, in-hospital death, and length of stay. DESIGN: A prospective cohort study. SETTING: The Service of General Surgery of a tertiary-care hospital. PATIENTS: A consecutive series of patients admitted for more than 1 day (N = 2,989). RESULTS: Sixty-two (2.1%) patients died and 503 (16.8%) acquired a nosocomial infection, of which 378 (12.6%) were surgical site and 44 (1.5%) were lower respiratory tract. Smoking (mainly past smoking) was associated with a worse health status (eg, longer preoperative stay and higher American Society of Anesthesiologists score). A long history of smoking (> or = 51 pack-years) increased postoperative admission to the intensive care unit (adjusted odds ratio [OR] = 2.86; 95% confidence interval [CI95], 1.21 to 6.77) and in-hospital mortality (adjusted OR = 2.56; CI95, 1.10 to 5.97). There was no relationship between current smoking and surgical-site infection (adjusted OR = 0.99; CI95, 0.72 to 1.35), whereas a relationship was observed between past smoking and surgical-site infection (adjusted OR = 1.46; CI95, 1.02 to 2.09). Current smoking and, to a lesser degree, past smoking augmented the risk of lower respiratory tract infection (adjusted OR = 3.21; CI95, 1.21 to 8.51). Smokers did not undergo additional surgical procedures more frequently during hospitalization. In the multivariate analysis, length of stay was similar for smokers and nonsmokers. CONCLUSION: Smoking increases in-hospital mortality, admission to the intensive care unit, and lower respiratory tract infection, but not surgical-site infection. Deleterious effects of smoking are also observed in past smokers and they cannot be counteracted by hospital cessation programs. PMID- 12558235 TI - Hemorrhage after coronary artery bypass graft procedures. AB - OBJECTIVES: To identify risk factors for excessive bleeding after coronary artery bypass graft (CABG) procedures and to quantify the outcomes related to this complication. DESIGN: We conducted a case-control study to identify risk factors for hemorrhage following CABG surgery and a historical cohort study to quantify outcomes of hemorrhage. SETTING: The cardiothoracic surgery service of a university hospital. RESULTS: Factors associated with excessive blood loss were recent catheterization (odds ratio [OR] = 0.44; 95% confidence interval [CI95], 0.21 to 0.91); age older than 65 years (OR = 1.94; CI95, 0.96 to 3.93); bypass time of 150 minutes or more (OR = 2.91; CI95, 1.09 to 7.81); and postoperative platelet count of 160,000/mm3 or less (OR = 2.36; CI95, 1.06 to 5.22). The attributable cost of a postoperative hemorrhage was $3,866 (P = .0002) overall, $9,912 (P = .0001) for patients who required reoperation, and $3,316 (P = .03) for those treated medically. The median attributable postoperative length of stay was 1 day longer for cases than for controls (P = .011). Postoperatively, patients who hemorrhaged received significantly larger volumes of packed red blood cells (P < .0001), fresh frozen plasma (P < .0001), platelets (P < .0001), plasminate (P = .007), protamine sulfate (P < .0001), desmopressin acetate (P < .0001), and epsilon-aminocaproic acid (P < .0001) than did controls. CONCLUSIONS: Age, duration of bypass, and postoperative platelet count were associated with excessive bleeding. Hemorrhage after CABG surgery significantly increased the length of stay and cost of care. PMID- 12558236 TI - An intranet-based automated system for the surveillance of nosocomial infections: prospective validation compared with physicians' self-reports. AB - OBJECTIVE: To examine the reliability of the data produced by an automated system for the surveillance of nosocomial infections. SETTING: A 906-bed, tertiary-care teaching hospital. DESIGN: Three surveillance techniques were concurrently performed in seven high-risk units during an 11-week period: automated surveillance (AS) based on the prospective processing of computerized medical records; laboratory-based ward surveillance (LBWS) based on the retrospective verification by ward clinicians of weekly reports of positive bacteriologic results; and a reference standard (RS) consisting of the infection control team reviewing case records of patients with positive bacteriology results. Bacteremia, urinary tract infections, and catheter-related infections were recorded for all inpatients. The performances (sensitivity, specificity, and time consumption) of AS and LBWS were compared with those of RS. RESULTS: Of 548 positive bacteriology samples included during the study period, 229 (42%) were classified as nosocomial infections. The overall sensitivity was 91% and 59% for AS and LBWS, respectively. The two methods had the same overall specificity value (91%). Kappa measures of agreement were 0.81 and 0.54 for AS and LBWS, respectively. AS required less time to collect data (54 seconds per week per unit) compared with LBWS (7 minutes and 43 seconds per week per unit) and RS (37 minutes and 15 seconds per week per unit). CONCLUSION: Our results confirm that the retrospective review of charts and laboratory data by physicians lacks sensitivity for the surveillance of nosocomial infections. The intranet-based automated method developed for this purpose was more accurate and less time consuming than the weekly, retrospective LBWS method. PMID- 12558237 TI - The prevalence and estimates of the cumulative incidence of hospital-acquired infections among patients admitted to Auckland District Health Board Hospitals in New Zealand. AB - OBJECTIVES: To report the pooled results of seven prevalence surveys of hospital acquired infections conducted between November 1996 and November 1999, and to use the data to predict the cumulative incidence of hospital-acquired infections in the same patient group. DESIGN: The summary and modeling of data gathered from the routine surveillance of the point prevalence of hospital-acquired infections. SETTING: Auckland District Health Board Hospitals (Auckland DHBH), the largest publicly funded hospital group in New Zealand supplying secondary and tertiary services. PATIENTS: All inpatients. METHOD: Point-prevalence surveys were conducted including all patients in Auckland DHBH. Standard definitions of hospital-acquired infection were used. The data from the seven surveys were pooled and used in a modeling exercise to predict the cumulative incidence of hospital-acquired infection. An existing method for the conversion of prevalence to cumulative incidence was applied. Results are presented for all patients and stratified by clinical service and site of hospital-acquired infection. RESULTS: The underlying patterns of hospital-acquired infection by site and service were stable during the seven time periods. The prevalence rate for all patients was 9.5%, with 553 patients identified with one or more hospital-acquired infections from a population of 5,819. The predicted cumulative incidence for all patients was 6.33% (95% confidence interval, 6.20% to 6.46%). CONCLUSIONS: The prevalence and the predicted cumulative incidence are similar to rates reported in the international literature. The validity of the predicted cumulative incidence derived here is not known. If it were accurate, then the application of this method would represent a cost-effective alternative to incidence studies. PMID- 12558238 TI - Resource utilization among patients with sepsis syndrome. AB - OBJECTIVE: To assess the resource utilization associated with sepsis syndrome in academic medical centers. DESIGN: Prospective cohort study. SETTING: Eight academic, tertiary-care centers. PATIENTS: Stratified random sample of 1,028 adult admissions with sepsis syndrome and all 248,761 other adult admissions between January 1993 and April 1994. The main outcome measures were length of stay (LOS) in total and after onset of sepsis syndrome (post-onset LOS) and total hospital charges. RESULTS: The mean LOS for patients with sepsis was 27.7 +/- 0.9 days (median, 20 days), with sepsis onset occurring after a mean of 8.1 +/- 0.4 days (median, 3 days). For all patients without sepsis, the LOS was 7.2 +/- 0.03 days (median, 4 days). In multiple linear regression models, the mean for patients with sepsis syndrome was 18.2 days, which was 11.0 days longer than the mean for all other patients (P < .0001), whereas the mean difference in total charges was $43,000 (both P < .0001). These differences were greater for patients with nosocomial as compared with community-acquired sepsis, although the groups were similar after adjusting for pre-onset LOS. Eight independent correlates of increased post-onset LOS and 12 correlates of total charges were identified. CONCLUSIONS: These data quantify the resource utilization associated with sepsis syndrome, and demonstrate that resource utilization is high in this group. Additional investigation is required to determine how much of the excess post onset LOS and charges are attributable to sepsis syndrome rather than the underlying medical conditions. PMID- 12558239 TI - Ethical treatment of animals in research. PMID- 12558240 TI - Virtual error sensing for active noise control in a one-dimensional waveguide: performance prediction versus measurement. PMID- 12558242 TI - Comment on "Development of panel loudspeaker system: design, evaluation and enhancement". PMID- 12558241 TI - Laboratory measurements of sound scattering from a buried sphere above and below the critical angle. PMID- 12558243 TI - Acoustic and vibration background noise in the collapsed structure of the World Trade Center. PMID- 12558244 TI - The role of envelope beat cues in the detection and discrimination of second order amplitude modulation. PMID- 12558245 TI - Central locus for nonspeech context effects on phonetic identification. PMID- 12558247 TI - Direct and inverse scattering of transient acoustic waves by a slab of rigid porous material. AB - This paper provides a temporal model of the direct and inverse scattering problem for the propagation of transient ultrasonic waves in a homogeneous isotropic slab of porous material having a rigid frame. This new time domain model of wave propagation takes into account the viscous and thermal losses of the medium as described by the model of Johnson et al. [D. L. Johnson, J. Koplik, and R. Dashen, J. Fluid. Mech. 176, 379 (1987)] and Allard [J. F. Allard (Chapman and Hall, London, 1993)] modified by a fractional calculus based method applied in the time domain. This paper is devoted to the analytical calculus of acoustic field in a slab of porous material. The main result is the derivation of the expression of the scattering operators (reflection and transmission) which are the responses of the medium to an incident acoustic pulse. In this model the reflection operator is the sum of two contributions: the first interface and the bulk of the medium. Experimental and numerical results are given as a validation of our model. PMID- 12558246 TI - Sounds produced by individual white whales, Delphinapterus leucas, from Svalbard during capture. PMID- 12558248 TI - The reflection of bounded inhomogeneous waves on a liquid/solid interface. AB - Fourier analysis and normal mode theory are used to describe the reflection of bounded inhomogeneous waves on a liquid/solid interface. Nonspecular reflection phenomena in the Rayleigh angle are studied in detail. In this way, an explanation is given for the Rayleigh dip phenomenon for positive inhomogeneity factors and the related result of a reflection coefficient larger than unity when the sign of the inhomogeneity factor is reversed. In the limit of large beamwidths, the reflection coefficient predicted by the infinite plane inhomogeneous wave theory is obtained. These results are entirely consistent with the experimental work published by Deschamps [J. Acoust. Soc. Am. 96, 2841-2848 (1994)]. The energy efficiency of Rayleigh wave excitation is investigated as well. It is shown that for large beamwidths, the energy efficiency for bounded inhomogeneous waves is considerably higher in comparison with Gaussian and square profiled beams. PMID- 12558249 TI - Experimental demonstration of noninvasive transskull adaptive focusing based on prior computed tomography scans. AB - Developing minimally invasive brain surgery by high-intensity focused ultrasound beams is of great interest in cancer therapy. However, the skull induces strong aberrations both in phase and amplitude, resulting in a severe degradation of the beam shape. Thus, an efficient brain tumor therapy would require an adaptive focusing, taking into account the effects of the skull. In this paper, we will show that the acoustic properties of the skull can be deduced from high resolution CT scans and used to achieve a noninvasive adaptive focusing. Simulations have been performed with a full 3-D finite differences code, taking into account all the heterogeneities inside the skull. The set of signals to be emitted in order to focus through the skull can thus be computed. The complete adaptive focusing procedure based on prior CT scans has been experimentally validated. This could have promising applications in brain tumor hyperthermia but also in transcranial ultrasonic imaging. PMID- 12558250 TI - Scattering from a single bubble near a roughened air-water interface: laboratory measurements and modeling. AB - The problem of scattering from a single bubble located close to a slightly roughened, air-water interface is studied both theoretically and experimentally. Two well-controlled laboratory experiments were performed to investigate the effects of surface roughness on the scattering response of the bubble. In the first experiment, a bubble of radius 1200 microm was placed on a fine thread at a variable distance, d, from the mean-still-water level of the surface, which was roughened using a wind source. In the second experiment, a bubble of radius 800 microm was utilized, while the water surface was roughened using a plunger wave making source. The waveheights and important characteristic length scales associated with each experiment were quantified using digital photography. The wind source produced waveheights that were represented by a Gaussian distribution, while the plunger source produced waveheights that were represented by a bimodal distribution. To model the acoustic measurements, an expression describing the four scattering paths, from source to bubble to receiver, was used. A random phase shift due to the surface roughness was added to the paths that interacted with the surface, and expectations of this phase shift were computed based on the analytical representations for the waveheight distribution. The data show good agreement with the simulations and the sensitivity of scattering from a subsurface bubble to small changes in waveheight is illustrated. The experiments highlight important parametric dependencies, which are summarized here, and the relation between monostatic and bistatic scattering is also discussed. PMID- 12558251 TI - Circumferential resonance modes of solid elastic cylinders excited by obliquely incident acoustic waves. AB - When an immersed solid elastic cylinder is insonified by an obliquely incident plane acoustic wave, some of the resonance modes of the cylinder are excited. These modes are directly related to the incidence angle of the insonifying wave. In this paper, the circumferential resonance modes of such immersed elastic cylinders are studied over a large range of incidence angles and frequencies and physical explanations are presented for singular features of the frequency incidence angle plots. These features include the pairing of one axially guided mode with each transverse whispering gallery mode, the appearance of an anomalous pseudo-Rayleigh in the cylinder at incidence angles greater than the Rayleigh angle, and distortional effects of the longitudinal whispering gallery modes on the entire resonance spectrum of the cylinder. The physical explanations are derived from Resonance Scattering Theory (RST), which is employed to determine the interior displacement field of the cylinder and its dependence on insonification angle. PMID- 12558252 TI - Sound source reconstruction using inverse boundary element calculations. AB - Whereas standard boundary element calculations focus on the forward problem of computing the radiated acoustic field from a vibrating structure, the aim in this work is to reverse the process, i.e., to determine vibration from acoustic field data. This inverse problem is brought on a form suited for solution by means of an inverse boundary element method. Since the numerical treatment of the inverse source reconstruction results in a discrete ill-posed problem, regularization is imposed to avoid unstable solutions dominated by errors. In the present work the emphasis is on Tikhonov regularization and parameter-choice methods not requiring an error-norm estimate for choosing the right amount of regularization. Several parameter-choice strategies have been presented lately, but it still remains to be seen how well these can handle industrial applications with real measurement data. In the present work it is demonstrated that the L-curve criterion is robust with respect to the errors in a real measurement situation. In particular, it is shown that the L-curve criterion is superior to the more conventional generalized cross-validation (GCV) approach for the present tire noise studies. PMID- 12558253 TI - Inverse estimation of the acoustic impedance of a porous woven hose from measured transmission coefficients. AB - A porous tube, comprised of a resin-coated woven fabric has recently been used as an effective component for use in intake systems of internal combustion engines to reduce the intake noise. For the prediction of the acoustic performance of an engine intake system with a porous woven hose, the acoustic wall impedance of the hose must be known. However, the accurate measurement of the wall impedance of a porous woven hose is not easy because of its peculiar acoustical and structural characteristics. A new measurement technique is proposed herein, that is valid over the low to mid frequency ranges. The acoustics impedance is inversely estimated from an overdetermined set of measured pressure transmission coefficients for specimens of different lengths and the reflection coefficient of end termination. The method involves only one measurement setup, and, as a result, it is very simple. A variation of the proposed method, an inverse estimation method using one of the four-pole parameters is also proposed. An error sensitivity analysis was performed to investigate the effect of measurement error on the accuracy of the final result. The measured TL for samples with arbitrary lengths and arbitrary porous frequency are in reasonably good agreement with values predicted from curve-fitted impedance data. PMID- 12558254 TI - Modeling of nonlinear ultrasound propagation in tissue from array transducers. AB - A computationally efficient model capable of simulating finite-amplitude ultrasound beam propagation in water and in tissue from phased linear arrays and other transducers of arbitrary quasiplanar geometry is described. It is based on a second-order operator splitting approach [Tavakkoli et al., J. Acoust. Soc. Am. 104, 2061-2072 (1998)], with a fractional step-marching scheme, whereby the effects of diffraction, attenuation, and nonlinearity can be computed independently over incremental steps. This approach is an extension to that of Christopher and Parker [J. Acoust. Soc. Am. 90, 507-521; 90, 488-499 (1991)], wherein linear and nonlinear effects are propagated separately over incremental steps, and the computation of the diffractive substeps are based on an angular spectrum technique with a modified sampling scheme for accurate and efficient implementation of diffractive propagation from nonradially symmetric sources. Results of the model are compared with published data. Predicted field profiles for nonlinear propagation in tissue from realistic array transducers using the pulse inversion method are presented. PMID- 12558255 TI - Acoustic streaming generated by standing waves in two-dimensional channels of arbitrary width. AB - An analytic solution is derived for acoustic streaming generated by a standing wave in a viscous fluid that occupies a two-dimensional channel of arbitrary width. The main restriction is that the boundary layer thickness is a small fraction of the acoustic wavelength. Both the outer, Rayleigh streaming vortices and the inner, boundary layer vortices are accurately described. For wide channels and outside the boundary layer, the solution is in agreement with results obtained by others for Rayleigh streaming. As channel width is reduced, the inner vortices increase in size relative to the Rayleigh vortices. For channel widths less than about 10 times the boundary layer thickness, the Rayleigh vortices disappear and only the inner vortices exist. The obtained solution is compared with those derived by Rayleigh, Westervelt, Nyborg, and Zarembo. PMID- 12558256 TI - Reynolds number effects on flow/acoustic mechanisms in spherical windscreens. AB - There is a practical need to fully understand the mechanisms involved in the flow/pressure fluctuations around a screened microphone. A stream of uniform flow with low-frequency turbulence encountering a rigid, impermeable spherical windscreen is considered in this study. Pressure distributions on the surface of the sphere are determined by the flow structure. Pressure fluctuations at the center of the sphere are then calculated based on the integration of surface pressure distributions. Because of the low-frequency assumption, results from steady-state laminar flows can be used to investigate the Reynolds number effects on wind noise reduction. Three types of flow have been studied in this paper: an inviscid case, a low-Reynolds-number Stokes flow, and intermediate- and high Reynolds-number flows. A Reynolds-number/wind-noise-reduction correlation shows that the wind noise reduction increases with decreasing Reynolds number. PMID- 12558257 TI - Acoustic near-field characteristics of a conical, premixed flame. AB - The occurrence of self-excited pressure oscillations routinely plagues the development of combustion systems. These oscillations are often driven by interactions between the flame and acoustic perturbations. This study was performed to characterize the structure of the acoustic field in the near field of the flame and the manner in which it is influenced by oscillation frequency, combustor geometry, flame length and temperature ratio. The results of these calculations indicate that the acoustic velocity has primarily one- and two dimensional features near the flame tip and base, respectively. The magnitude of the radial velocity components increases with temperature ratio across the flame, while their axial extent increases with frequency. However, the acoustic pressure has primarily one-dimensional characteristics. They also show that the acoustic field structure exhibits only moderate dependencies upon area expansion and flame temperature ratio for values typical of practical systems. Finally, they show that the local characteristics of the acoustic field, as well as the overall plane-wave reflection coefficient, exhibit a decreasing dependence upon the flame length as the area expansion ratio increases. PMID- 12558258 TI - Application of an acoustic noise removal method to aircraft-based atmospheric temperature measurements. AB - An acoustic noise removal method is used to reject engine acoustical disturbances from aircraft-based atmospheric temperature measurements. Removal of engine noise from atmospheric temperature measurements allows a larger wave number range to be fit while quantifying the magnitude of atmospheric temperature turbulence. The larger wave number range was found to result in a more statistically certain spectral slope estimate, with up to a 50% reduction in the standard deviation of measured spectral slopes. The noise removal technique was found to break down under conditions of weak atmospheric temperature turbulence where the engine acoustical disturbance can be several orders of magnitude larger than atmospheric temperature turbulence. PMID- 12558259 TI - Active stereo sound localization. AB - Estimating the direction of arrival of sound in three-dimensional space is typically performed by generalized time-delay processing on a set of signals from a fixed array of omnidirectional microphones. This requires specialized multichannel A/D hardware, and careful arrangement of the microphones into an array. This work is motivated by the desire to instead only use standard two channel audio A/D hardware and portable equipment. To estimate direction of arrival of persistent sound, the position of the microphones is made variable by mounting them on one or more computer-controlled pan-and-tilt units. In this paper, we describe the signal processing and control algorithm of a device with two omnidirectional microphones on a fixed baseline and two rotational degrees of freedom. Experimental results with real data are reported with both impulsive and speech sounds in an untreated, normally reverberant indoor environment. PMID- 12558260 TI - A method of images for a penetrable acoustic waveguide. AB - In this paper the complex-image approximation to the reflection coefficient for water over a seabed half-space is used to generate an image representation for a bounded acoustic waveguide with an underlying layered seabed. The images are true point sources; they have constant amplitudes which are raypath independent and, in the case of a Pekeris waveguide, frequency-independent. This image representation is ideal for constructing the Green's function kernel of the boundary integral equation method for target scattering in a waveguide. The singular behavior of the Green's function for an infinitesimal source/receiver separation, possibly with the target adjacent to one of the interfaces, is modeled correctly and the image expansion has a simple analytic form which can be analytically differentiated. The method is also accurate for significant source/receiver separations, which means that it can be used in the modeling of scattering from large-sized objects and can also be used as an efficient and accurate short-range propagation model for harmonic and broadband propagation in a penetrable waveguide. PMID- 12558261 TI - Broadband sound propagation in shallow water and geoacoustic inversion. AB - Part of an experiment to test a measurement package in a shallow water region in the Gulf of Mexico was designed to gather broadband acoustic data suitable for inversion to estimate seabed geoacoustic parameters. Continuous wave tow acoustic signals at multiple frequencies and broadband impulsive source signals were recorded on a horizontal line array in a high-noise environment. Simulated annealing with a normal mode forward propagation model is utilized to invert for a geoacoustic representation of the seabed. Several inversions are made from different data samples of two light bulb implosions, the measured sound speed profiles at the HLA and at the positions of the light bulb deployments, and for two different cost functions. The different cost functions, measured sound speed profiles, and measured time series result in different inverted geoacoustic profiles from which transmission loss is generated for comparison with measurements. On the basis of physical consistency and from the comparison of the transmission loss and time series, a best estimate geoacoustic profile is selected and compared to those obtained from previously reported inversions. Uncertainties in the sound speed profile are shown to affect the uncertainties of the estimated seabed parameters. PMID- 12558262 TI - Spectral and modal formulations for the Doppler-shifted field scattered by an object moving in a stratified medium. AB - Spectral and normal mode formulations for the three-dimensional field scattered by an object moving in a stratified medium are derived using full-field wave theory. The derivations are based on Green's theorem for the time-domain scalar wave equation and account for Doppler effects induced by target motion as well as source and receiver motion. The formulations are valid when multiple scattering between the object and waveguide boundaries can be neglected, and the scattered field can be expressed as a linear function of the object's plane wave scattering function. The advantage of the spectral formulation is that it incorporates the entire wave number spectrum, including evanescent waves, and therefore can potentially be used at much closer ranges to the target than the modal formulation. The normal mode formulation is more computationally efficient but is limited to longer ranges. For a monochromatic source that excites N incident modes in the waveguide, there will be roughly N2 distinct harmonic components in the scattered field. The Doppler shifts in the scattered field are highly dependent upon the waveguide environment, target shape, and measurement geometry. The Doppler effects are illustrated through a number of canonical examples. PMID- 12558263 TI - Motion compensation for adaptive horizontal line array processing. AB - Large aperture horizontal line arrays have small resolution cells and can be used to separate a target signal from an interference signal by array beamforming. High-resolution adaptive array processing can be used to place a null at the interference signal so that the array gain can be much higher than that of conventional beamforming. But these nice features are significantly degraded by the source motion, which reduces the time period under which the environment can be considered stationary from the array processing point of view. For adaptive array processing, a large number of data samples are generally required to minimize the variance of the cross-spectral density, or the covariance matrix, between the array elements. For a moving source and interference, the penalty of integrating over a large number of samples is the spread of signal and interference energy to more than one or two eigenvalues. The signal and interference are no longer clearly identified by the eigenvectors and, consequently, the ability to suppress the interference suffers. We show in this paper that the effect of source motion can be compensated for the (signal) beam covariance matrix, thus allowing integration over a large number of data samples without loss in the signal beam power. We employ an equivalent of a rotating coordinate frame to track the signal bearing change and use the waveguide invariant theory to compensate the signal range change by frequency shifting. PMID- 12558264 TI - Narrow band laser-generated surface acoustic waves using a formed source in the ablative regime. AB - A narrow band laser-generated acoustic signal was created using a 4-element lenticular array. This arrangement of the array produces an acoustical signal with frequency content that is compatible with the response of a noncontact and remote broadband receiver, such as a capacitive air-coupled transducer. To support the experimental observations, a simplified concept is presented to explain the effect of a line array source on the frequency content of a surface acoustic wave. The analytical model solution for the wave front shape is derived from the point load solution of Lamb's problem that represents the displacement of a surface acoustic wave generated by an ablative line array. The distribution function, which was used for the model to represent the laser light energy, was tailored to depict the actual energy distribution that illuminates the lenticular array. Filtering functions are applied to the resultant surface displacement function to retain frequencies similar to those detected by the broadband 50 kHz 2.25 MHz receiver. The theoretical model showed good agreement with experimental results. PMID- 12558265 TI - Derivation of an optimal directivity pattern for sweet spot widening in stereo sound reproduction. AB - In this paper the correction of the degradation of the stereophonic illusion during sound reproduction due to off-center listening is investigated. The main idea is that the directivity pattern of a loudspeaker array should have a well defined shape such that a good stereo reproduction is achieved in a large listening area. Therefore, a mathematical description to derive an optimal directivity pattern l(opt) that achieves sweet spot widening in a large listening area for stereophonic sound applications is described. This optimal directivity pattern is based on parametrized time/intensity trading data coming from psycho acoustic experiments within a wide listening area. After the study, the required digital FIR filters are determined by means of a least-squares optimization method for a given stereo base setup (two pair of drivers for the loudspeaker arrays and 2.5-m distance between loudspeakers), which radiate sound in a broad range of listening positions in accordance with the derived l(opt). Informal listening tests have shown that the l(opt) worked as predicted by the theoretical simulations. They also demonstrated the correct central sound localization for speech and music for a number of listening positions. This application is referred to as "Position-Independent (PI) stereo." PMID- 12558266 TI - Electromechanical coupling factor of capacitive micromachined ultrasonic transducers. AB - Recently, a linear, analytical distributed model for capacitive micromachined ultrasonic transducers (CMUTs) was presented, and an electromechanical equivalent circuit based on the theory reported was used to describe the behavior of the transducer [IEEE Trans. Ultrason. Ferroelectr. Freq. Control 49, 159-168 (2002)]. The distributed model is applied here to calculate the dynamic coupling factor k(w) of a lossless CMUT, based on a definition that involves the energies stored in a dynamic vibration cycle, and the results are compared with those obtained with a lumped model. A strong discrepancy is found between the two models as the bias voltage increases. The lumped model predicts an increasing dynamic k factor up to unity, whereas the distributed model predicts a more realistic saturation of this parameter to values substantially lower. It is demonstrated that the maximum value of k(w), corresponding to an operating point close to the diaphragm collapse, is 0.4 for a CMUT single cell with a circular membrane diaphragm and no parasitic capacitance (0.36 for a cell with a circular plate diaphragm). This means that the dynamic coupling factor of a CMUT is comparable to that of a piezoceramic plate oscillating in the thickness mode. Parasitic capacitance decreases the value of k(w), because it does not contribute to the energy conversion. The effective coupling factor k(eff) is also investigated, showing that this parameter coincides with k(w) within the lumped model approximation, but a quite different result is obtained if a computation is made with the more accurate distributed model. As a consequence, k(eff), which can be measured from the transducer electrical impedance, does not give a reliable value of the actual dynamic coupling factor. PMID- 12558267 TI - A directional acoustic array using silicon micromachined piezoresistive microphones. AB - The need for noise source localization and characterization has driven the development of advanced sound field measurement techniques using microphone arrays. Unfortunately, the cost and complexity of these systems currently limit their widespread use. Directional acoustic arrays are commonly used in wind tunnel studies of aeroacoustic sources and may consist of hundreds of condenser microphones. A microelectromechanical system (MEMS)-based directional acoustic array system is presented to demonstrate key technologies to reduce the cost, increase the mobility, and improve the data processing efficiency versus conventional systems. The system uses 16 hybrid-packaged MEMS silicon piezoresistive microphones that are mounted to a printed circuit board. In addition, a high-speed signal processing system was employed to generate the array response in near real time. Dynamic calibrations of the microphone sensor modules indicate an average sensitivity of 831 microV/Pa with matched magnitude (+/-0.6 dB) and phase (+/-1 degree) responses between devices. The array system was characterized in an anechoic chamber using a monopole source as a function of frequency, sound pressure level, and source location. The performance of the MEMS based array is comparable to conventional array systems and also benefits from significant cost savings. PMID- 12558268 TI - Acoustic scattering by a cylindrical shell with symmetric line constraints in the heavy fluid-loading limit. AB - A cylindrical shell, modelled using Donnell-Mushtari thin shell theory, is reinforced by two internal rigid plates attached to the shell along lines parallel to the shell axis. A circumferential mode expansion is used to obtain numerical results for the scattered sound field due to the presence of the reaction forces along the attachment lines. In the heavy fluid-loading limit, which is appropriate for low and mid-frequency ranges for practical underwater structures, asymptotic analysis is presented which allows the peak frequencies in the scattered field due to the reaction forces to be expressed (to leading order) in terms of the geometry and the shell and fluid parameters. These predictions agree well with results obtained by numerically evaluating the infinite sums needed to calculate the reaction forces and hence the scattered field. PMID- 12558269 TI - Leaky helical flexural wave backscattering contributions from tilted water-filled cylindrical shells. AB - Helical flexural waves on a bluntly truncated tilted water-filled cylindrical steel shell in water are found to give large contributions to the backscattering above the coincidence frequency. The presence of the water inside the shell increases the damping of the leaky wave when short tone bursts are used. The magnitude of the scattering is found by modifying a ray analysis developed for empty shells. When longer bursts are used, some of the internally radiated energy (corresponding to the case of one internal chord) is superposed on the ordinary helical ray backscattering. This occurs as a consequence of the internal excitation of helical rays. PMID- 12558270 TI - Application of Pade via Lanczos approximations for efficient multifrequency solution of Helmholtz problems. AB - This paper addresses the efficient solution of acoustic problems in which the primary interest is obtaining the solution only on restricted portions of the domain but over a wide range of frequencies. The exterior acoustics boundary value problem is approximated using the finite element method in combination with the Dirichlet-to-Neumann (DtN) map. The restriction domain problem is formally posed in transfer function form based on the finite element solution. In order to obtain the solution over a range of frequencies, a matrix-valued Pade approximation of the transfer function is employed, using a two-sided block Lanczos algorithm. This approach provides a stable and efficient representation of the Pade approximation. In order to apply the algorithm, it is necessary to reformulate the transfer function due to the frequency dependency in the nonreflecting boundary condition. This is illustrated for the case of the DtN boundary condition, but there is no restriction on the approach which can also be applied to other radiation boundary conditions. Numerical tests confirm that the approach offers significant computational speed-up. PMID- 12558271 TI - A passive means for cancellation of structurally radiated tones. AB - The concept of cancellation of constant-frequency sound radiated from a vibrating surface by means of an attached mechanical oscillator is discussed. It is observed that the mass of a mechanical oscillator whose spring is attached to the vibrating surface will vibrate at comparatively large amplitudes and out of phase with that surface, provided that the surface vibrates at a frequency that is slightly higher than the oscillator's natural frequency. From this observation it is concluded that an oscillator's mass with a relatively small surface area can produce a volume velocity that is equal and opposite to that of the vibrating surface, resulting in cancellation of the sound radiated from the surface. Practical considerations in the design of such an oscillator are discussed, and the canceling performance from oscillators consisting of edge-supported circular disks is analyzed. An experimental canceling oscillator consisting of an edge supported disk is described, and measurements made with this disk attached to a piston are shown to be in good agreement with analytical predictions. A tonal noise reduction exceeding 20 dB was demonstrated experimentally. PMID- 12558272 TI - Lattice form adaptive infinite impulse response filtering algorithm for active noise control. AB - In some situations of active noise control, infinite impulse response (IIR) filters are more suitable than finite impulse response (FIR) filters owing to the poles in the transfer function. A number of algorithms have been derived for applying IIR filters in active noise control; however, most of them use the direct form IIR filter structure, which faces the difficulties of checking stability and relatively slow convergence speed for noise composed of narrow-band components with large power disparity. To overcome these difficulties along with using the direct form IIR filters, a new adaptive algorithm is proposed in this paper, which uses and updates the lattice form adaptive IIR filter in an active noise control system. Full mathematical derivations of the proposed algorithm are presented, and the comparison between the proposed algorithm and the commonly used filtered-u LMS and filtered-v LMS algorithms shows the superiority of the proposed algorithm. PMID- 12558273 TI - A- and C-weighted sound levels as predictors of the annoyance caused by shooting sounds, for various facade attenuation types. AB - In a previous study on the annoyance caused by a great variety of shooting sounds [J. Acoust. Soc. Am. 109, 244-253 (2001)], it was shown that the annoyance, as rated indoors with the windows closed, could be adequately predicted from the outdoor A-weighted and C-weighted sound-exposure levels [ASEL (L(AE)) and CSEL (L(CE))] of the impulse sounds. The explained variance in the mean ratings by (outdoor) ASEL was significantly increased by adding the product (L(CE) - L(AE))(L(AE)) as a second variable. In the present study it was investigated to which extent the additional contribution of the second predictor is also relevant for facade attenuation types with lower and higher degrees of sound isolation than applied previously. Twenty subjects rated the indoor annoyance caused by 11 different impulse types produced by firearms ranging in caliber from 7.62 to 155 mm, at various levels and for five facade attenuation conditions. The effect of facade attenuation on the ratings was large and consistent. In all conditions, an optimal prediction of the annoyance was obtained with outdoor ASEL as the first, and (L(CE) - L(AE))(L(AE)) as the second predictor. The benefit of the second predictor, expressed as the increase in the explained variance, ranged from 2.5 to 55 percent points, and strongly increased with the degree of facade attenuation. It was concluded that for the determination of the rating sound level, the acoustic parameters ASEL and CSEL are very powerful. In addition, the results showed that for the whole set of impulses included, the annoyance could also be predicted very well by the weighted sum of indoor ASEL and the product (L(CE) - L(AE))(L(AE)). PMID- 12558274 TI - Investigation of phase coupling phenomena in sustained portion of musical instruments sound. AB - This work investigates aperiodicities that occur in the sustained portion of a sound of musical instrument played by a human player, due to synchronous versus asynchronous deviations of the partial phases. By using an additive sinusoidal analysis, phases of individual partials are precisely extracted and their correlation statistics and coupling effects are analyzed. It is shown that various musical instruments exhibit different phase coupling characteristics. The effect of phase coupling is compared to analysis by means of higher order statistics and it is shown that both methods are closely mathematically related. Following a detailed analysis of phase coupling for various musical instruments it is suggested that phase coupling is an important characteristic of a sustained portion of sound of individual musical instruments, and possibly even of instrumental families. Interesting differences in phase deviations where found for the flute, trumpet and cello. For the cello, the effect of vibrato is examined by comparing the analysis of a closed string sound played with a natural vibrato to analysis of an open string sound that contains no vibrato. Following, a possible model for phase deviations in the cello is presented and a simulation of phase fluctuations for this model is performed. PMID- 12558275 TI - Compensation for source nonstationarity in multireference, scan-based near-field acoustical holography. AB - Multireference, scan-based near-field acoustical holography is a useful measurement tool that can be applied when an insufficient number of microphones is available to make measurements on a complete hologram surface simultaneously. The scan-based procedure can be used to construct a complete hologram by joining together subholograms captured using a relatively small, roving scan array and a fixed reference array. For the procedure to be successful, the source levels must remain stationary for the time taken to record the complete hologram; that is unlikely to be the case in practice, however. Usually, the reference signal levels measured during each scan differ from each other with the result that spatial noise is added to the hologram. A procedure to suppress the effects of source level, and hence reference level, variations is proposed here. The procedure is based on a formulation that explicitly features the acoustical transfer functions between the sources and both the reference and scanning, field microphones. When it is assumed that source level changes do not affect the sources' directivity, a nonstationarity compensation procedure can be derived that is based on measured transfer functions between the reference and field microphones. It has been verified both experimentally and in numerical simulations that the proposed procedure can help suppress spatially distributed noise caused by the type of source level nonstationarity that is characteristic of realistic sources. PMID- 12558276 TI - An auditory-periphery model of the effects of acoustic trauma on auditory nerve responses. AB - Acoustic trauma degrades the auditory nerve's tonotopic representation of acoustic stimuli. Recent physiological studies have quantified the degradation in responses to the vowel /E/ and have investigated amplification schemes designed to restore a more correct tonotopic representation than is achieved with conventional hearing aids. However, it is difficult from the data to quantify how much different aspects of the cochlear pathology contribute to the impaired responses. Furthermore, extensive experimental testing of potential hearing aids is infeasible. Here, both of these concerns are addressed by developing models of the normal and impaired auditory peripheries that are tested against a wide range of physiological data. The effects of both outer and inner hair cell status on model predictions of the vowel data were investigated. The modeling results indicate that impairment of both outer and inner hair cells contribute to degradation in the tonotopic representation of the formant frequencies in the auditory nerve. Additionally, the model is able to predict the effects of frequency-shaping amplification on auditory nerve responses, indicating the model's potential suitability for more rapid development and testing of hearing aid schemes. PMID- 12558277 TI - Ear-canal acoustic admittance and reflectance effects in human neonates. I. Predictions of otoacoustic emission and auditory brainstem responses. AB - This report describes the extent to which ear-canal acoustic admittance and energy reflectance (YR) in human neonates (1) predict otoacoustic emission (OAE) levels and auditory brainstem response (ABR) latencies, and (2) classify OAE and ABR responses as present or absent. Analyses are reported on a subset of ears in which hearing screening measurements were obtained previously [Norton et al., Ear. Hear. 21, 348-356 (2000a)]. Tests on 1405 ears included YR, distortion product OAEs, transient-evoked OAEs, and ABR. Principal components analysis reduced the 33 YR variables to 5-7 factors. OAE levels decreased and ABR latencies increased with increasing high-frequency energy reflectance. Up to 28% of the variance in OAE levels and 12% of the variance in ABR wave-V latencies were explained by these factors. Thus, the YR response indirectly encodes information on inter-ear variations in forward and reverse middle-ear transmission. The YR factors classify OAEs with an area under the relative operating characteristic (ROC) curve as high as 0.79, suggesting that middle-ear dysfunction is partly responsible for the inability to record OAEs in some ears. The YR factors classified ABR responses less well, with ROC areas of 0.64 for predicting wave-V latency and 0.56 for predicting Fsp. PMID- 12558279 TI - Objective estimates of cochlear tuning by otoacoustic emission analysis. AB - A new method is presented for estimating cochlear tuning starting from measurements of either the transient evoked otoacoustic emission latency or the spontaneous otoacoustic emission minimal spacing. This method could be useful in obtaining indirect information about the tuning curve, particularly for subjects that, like neonates, cannot be studied with psycho-acoustical techniques. Theoretical models of the acoustic transmission along the cochlea based on the transmission line formalism predict a relation between the otoacoustic emission latency and the frequency. This relation depends on the tuning curve, i.e., the frequency dependence of the quality factor of the cochlear resonances. On the other hand, models for the generation of spontaneous emissions based on the concept of coherent scattering from cochlear random inhomogeneities imply an independent relation between the tuning curve and the minimal frequency spacing between spontaneous emissions. In this study, experimental measurements of the otoacoustic emission latency and of the minimal spacing between spontaneous emissions are presented. Theoretical relations are derived, which connect these two measured quantities and the tuning curve. The typically longer latency of neonates implies a higher degree of tuning at high levels of stimulation. PMID- 12558278 TI - Ear-canal acoustic admittance and reflectance measurements in human neonates. II. Predictions of middle-ear in dysfunction and sensorineural hearing loss. AB - This report describes relationships between middle-ear measurements of acoustic admittance and energy reflectance (YR) and measurements of hearing status using visual reinforcement audiometry in a neonatal hearing-screening population. Analyses were performed on 2638 ears in which combined measurements were obtained [Norton et al., Ear Hear. 21, 348-356 (2000)]. The measurements included distortion-product otoacoustic emissions (DPOAE), transient evoked otoacoustic emissions (TEOAE), and auditory brainstem responses (ABR). Models to predict hearing status using DPOAEs, TEOAEs, or ABRs were each improved by the addition of the YR factors as interactions, in which factors were calculated using factor loadings from Keefe et al. [J. Acoust. Soc. Am. 113, 389-406 (2003)]. This result suggests that information on middle-ear status improves the ability to predict hearing status. The YR factors were used to construct a middle-ear dysfunction test on 1027 normal-hearing ears in which DPOAE and TEOAE responses were either both present or both absent, the latter condition being viewed as indicative of middle-ear dysfunction. The middle-ear dysfunction test classified these ears with a nonparametric area (A) under the relative operating characteristic curve of A = 0.86, and classified normal-hearing ears that failed two-stage hearing screening tests with areas A = 0.84 for DPOAE/ABR, and A = 0.81 for TEOAE/ABR tests. The middle-ear dysfunction test adequately generalized to a new sample population (A = 0.82). PMID- 12558280 TI - Physiopathological significance of distortion-product otoacoustic emissions at 2f1-f2 produced by high- versus low-level stimuli. AB - Distortion product otoacoustic emissions emitted by the cochlea at 2f1-f2 in response to pairs of pure tones at f1 and f2 (DPOAE) form a class of otoacoustic emissions and as such, are viewed as a reliable tool for screening outer hair cell (OHC) dysfunctions on a pass/fail basis. However, the persistence of residual DPOAEs from impaired cochleae at high stimulus levels has suggested that above 60-70 dB SPL, instead of reflecting "active" cochlear motion, DPOAEs might represent another "passive" modality: they would thus become unsuitable for analyzing cochlear function. The present work reports the consequences on high- vs low-level DPOAEs of three types of cochlear impairments involving OHCs: progressive OHC degeneration of genetic origin in CD1 mice, complete cochlear ischemia in gerbils, and furosemide injection vs ischemia-reperfusion in gerbils. An alternative to the "active-passive" model was used wherein regardless of stimulus level, cubic DPOAEs are produced by N (probably OHC-borne) nonlinear elements driven by input I and modulated by a function F3 of their operating point o; thus, DPOAE proportional to NI3F3(o). When OHCs degenerated, thereby implying a decrease of N, DPOAE levels also decreased regardless of the stimulus level up to 80 dB SPL, in line with the previous formula but at variance with the prediction of the active-passive concept. Instead of affecting N, the other two experiments impaired the efficiency of the cochlear feedback loop as a result of its electrical drive being decreased by strial dysfunction. As it is well accepted that the impaired basilar-membrane motion, although greatly reduced at low levels, tends to catch up with a normal one at higher levels, it was assumed the same was true with I so that DPOAE levels had to be, and indeed were little affected at high levels while plummeting at low levels, without any need for invoking two modalities for DPOAE generation. Finally, comparisons of furosemide vs ischemia effects revealed additional influences on DPOAEs, possibly accounted for by function F3(o). These results lead to the proposal that although high level DPOAEs are expected to be poor audiometric indicators, they seem well adapted to assessing the functional integrity of nonlinear elements in OHCs, i.e., presumably their mechanoelectrical transduction channels. PMID- 12558281 TI - Effect of current stimulus on in vivo cochlear mechanics. AB - In this paper, the influence of direct current stimulation on the acoustic impulse response of the basilar membrane (BM) is studied. A positive current applied in the scala vestibuli relative to a ground electrode in the scala tympani is found to enhance gain and increase the best frequency at a given location on the BM. An opposite effect is found for a negative current. Also, the amplitude of low-frequency cochlear microphonic at high sound levels is found to change with the concurrent application of direct current stimulus. BM vibrations in response to pure tone acoustic excitation are found to possess harmonics whose levels relative to the fundamental increase with the application of positive current and decrease with the application of negative current. A model for outer hair cell activity that couples changes in length and stiffness to transmembrane potential is used to interpret the results of these experiments and others in the literature. The importance of the in vivo mechanical and electrical loading is emphasized. Simulation results show the somewhat paradoxical finding that for outer hair cells under tension, hyperpolarization causes shortening of the cell length due to the dominance of voltage dependent stiffness changes. PMID- 12558282 TI - Effect of outer hair cell piezoelectricity on high-frequency receptor potentials. AB - The low-pass voltage response of outer hair cells predicted by conventional equivalent circuit analysis would preclude the active force production at high frequencies. We have found that the band pass characteristics can be improved by introducing the piezoelectric properties of the cell wall. In contrast to the conventional analysis, the receptor potential does not tend to zero and at any frequency is greater than a limiting value. In addition, the phase shift between the transduction current and receptor potential tends to zero. The piezoelectric properties cause an additional, strain-dependent, displacement current in the cell wall. The wall strain is estimated on the basis of a model of the cell deformation in the organ of Corti. The limiting value of the receptor potential depends on the ratio of a parameter determined by the piezoelectric coefficients and the strain to the membrane capacitance. In short cells, we have found that for the low-frequency value of about 2-3 mV and the strain level of 0.1% the receptor potential can reach 0.4 mV throughout the whole frequency range. In long cells, we have found that the effect of the piezoelectric properties is much weaker. These results are consistent with major features of the cochlear amplifier. PMID- 12558283 TI - Effect of amplitude modulation coherence for masked speech signals filtered into narrow bands. AB - Introduction of masker amplitude modulation (AM) can improve signal detection in a number of paradigms. In some cases this advantage depends on the coherence of modulation across a relatively wide frequency range. In the experiments described below, observers were asked to identify masked spondee words produced by a single male talker. The target spondees and masking noise were filtered into nine narrow bands, and the coherence of AM of either the speech signal or noise masker was manipulated. Inherent modulation of the masker bands was manipulated via assignment of real and imaginary values to the associated components of each band in the frequency domain, and AM of speech bands was achieved via multiplication with envelopes extracted from these maskers. Responses were based on two alternatives, four alternatives, or open response sets. The effect of masker AM coherence was highly dependent upon the size of the response set: coherent AM was associated with better thresholds in a two-alternative response set, but poorer thresholds in an open response set. Results with AM speech did not depend critically upon the across-frequency temporal synchrony of AM imposed on the speech material. PMID- 12558284 TI - Detection and direction-discrimination of diotic and dichotic ramp modulations in amplitude and phase. AB - When the source of a tone moves with respect to a listener's ears, dichotic (or interaural) phase and amplitude modulations (PM and AM) are produced. Two experiments investigated the psychophysical characteristics of dichotic linear ramp modulations in phase and amplitude, and compared them with the psychophysics of diotic PM and AM. In experiment 1, subjects were substantially more sensitive to dichotic PM than diotic PM, but AM sensitivity was equivalent in the dichotic and diotic conditions. Thresholds for discriminating modulation direction were smaller than detection thresholds for dichotic AM, and both diotic AM and PM. Dichotic PM discrimination thresholds were similar to detection thresholds. In experiment 2, the effects of ramp duration were examined. Sensitivity to dichotic AM and PM, and diotic AM increased as duration was increased from 20 ms to 200 ms. The functions relating sensitivity to ramp duration differed across the stimuli; sensitivity to dichotic PM increased more rapidly than sensitivity to dichotic or diotic AM. This was also reflected in shorter time-constants and minimum integration times for dichotic PM detection. These findings support the hypothesis that the analysis of dichotic PM and AM rely on separate mechanisms. PMID- 12558285 TI - Analysis of the three-dimensional tongue shape using a three-index factor analysis model. AB - Three-dimensional tongue shape during vowel production is analyzed using the three-mode PARAFAC (parallel factors) model. Three-dimensional MRI images of five speakers (9 vowels) are analyzed. Sixty-five virtual fleshpoints (13 segments along the rostral-caudal dimension and 5 segments along the right-left direction) are chosen based on the interpolated tongue shape images. Methods used to adjust the alignment of MRI images, to set up the fleshpoints, and to measure the position of the fleshpoints are presented. PARAFAC analysis of this 3D coordinate data results in a stable two-factor solution that explains about 70% of the variance. PMID- 12558286 TI - Flow visualization and pressure distributions in a model of the glottis with a symmetric and oblique divergent angle of 10 degrees. AB - Modeling the human larynx can provide insights into the nature of the flow and pressures within the glottis. In this study, the intraglottal pressures and glottal jet flow were studied for a divergent glottis that was symmetric for one case and oblique for another. A Plexiglas model of the larynx (7.5 times life size) with interchangeable vocal folds was used. Each vocal fold had at least 11 pressure taps. The minimal glottal diameter was held constant at 0.04 cm. The glottis had an included divergent angle of 10 degrees. In one case the glottis was symmetric. In the other case, the glottis had an obliquity of 15 degrees. For each geometry, transglottal pressure drops of 3, 5, 10, and 15 cm H2O were used. Pressure distribution results, suggesting significantly different cross-channel pressures at glottal entry for the oblique case, replicate the data in another study by Scherer et al. [J. Acoust. Soc. Am. 109, 1616-1630 (2001b)]. Flow visualization using a LASER sheet and seeded airflow indicated separated flow inside the glottis. Separation points did not appear to change with flow for the symmetric glottis, but for the oblique glottis moved upstream on the divergent glottal wall as flow rate increased. The outgoing glottal jet was skewed off-axis for both the symmetric and oblique cases. The laser sheet showed asymmetric circulating regions in the downstream region. The length of the laminar core of the glottal jet was less than approximately 0.6 cm, and decreased in length as flow increased. The results suggest that the glottal obliquity studied here creates significantly different driving forces on the two sides of the glottis (especially at the entrance to the glottis), and that the skewed glottal jet characteristics need to be taken into consideration for modeling and aeroacoustic purposes. PMID- 12558287 TI - The synergy between speech production and perception. AB - Speech intelligibility is known to be relatively unaffected by certain deformations of the acoustic spectrum. These include translations, stretching or contracting dilations, and shearing of the spectrum (represented along the logarithmic frequency axis). It is argued here that such robustness reflects a synergy between vocal production and auditory perception. Thus, on the one hand, it is shown that these spectral distortions are produced by common and unavoidable variations among different speakers pertaining to the length, cross sectional profile, and losses of their vocal tracts. On the other hand, it is argued that these spectral changes leave the auditory cortical representation of the spectrum largely unchanged except for translations along one of its representational axes. These assertions are supported by analyses of production and perception models. On the production side, a simplified sinusoidal model of the vocal tract is developed which analytically relates a few "articulatory" parameters, such as the extent and location of the vocal tract constriction, to the spectral peaks of the acoustic spectra synthesized from it. The model is evaluated by comparing the identification of synthesized sustained vowels to labeled natural vowels extracted from the TIMIT corpus. On the perception side a "multiscale" model of sound processing is utilized to elucidate the effects of the deformations on the representation of the acoustic spectrum in the primary auditory cortex. Finally, the implications of these results for the perception of generally identifiable classes of sound sources beyond the specific case of speech and the vocal tract are discussed. PMID- 12558288 TI - Effects of prosodic boundary on /aC/ sequences: acoustic results. AB - This study presents various acoustic measures used to examine the sequence /a # C/, where "#" represents different prosodic boundaries in French. The 6 consonants studied are /b d g f s S/ (3 stops and 3 fricatives). The prosodic units investigated are the utterance, the intonational phrase, the accentual phrase, and the word. It is found that vowel target values, formant transitions into the stop consonant, and the rate of change in spectral tilt into the fricative, are affected by the strength of the prosodic boundary. F1 becomes higher for /a/ the stronger the prosodic boundary, with the exception of one speaker's utterance data, which show the effects of articulatory declension at the utterance level. Various effects of the stop consonant context are observed, the most notable being a tendency for the vowel /a/ to be displaced in the direction of the F2 consonant "locus" for /d/ (the F2 consonant values for which remain relatively stable across prosodic boundaries) and for /g/ (the F2 consonant values for which are displaced in the direction of the velar locus in weaker prosodic boundaries, together with those of the vowel). Velocity of formant transition may be affected by prosodic boundary (with greater velocity at weaker boundaries), though results are not consistent across speakers. There is also a tendency for the rate of change in spectral tilt moving from the vowel to the fricative to be affected by the presence of a prosodic boundary, with a greater rate of change at the weaker prosodic boundaries. It is suggested that spectral cues, in addition to duration, amplitude, and F0 cues, may alert listeners to the presence of a prosodic boundary. PMID- 12558289 TI - Learning to produce speech with an altered vocal tract: the role of auditory feedback. AB - Modifying the vocal tract alters a speaker's previously learned acoustic articulatory relationship. This study investigated the contribution of auditory feedback to the process of adapting to vocal-tract modifications. Subjects said the word /tas/ while wearing a dental prosthesis that extended the length of their maxillary incisor teeth. The prosthesis affected /s/ productions and the subjects were asked to learn to produce "normal" /s/'s. They alternately received normal auditory feedback and noise that masked their natural feedback during productions. Acoustic analysis of the speakers' /s/ productions showed that the distribution of energy across the spectra moved toward that of normal, unperturbed production with increased experience with the prosthesis. However, the acoustic analysis did not show any significant differences in learning dependent on auditory feedback. By contrast, when naive listeners were asked to rate the quality of the speakers' utterances, productions made when auditory feedback was available were evaluated to be closer to the subjects' normal productions than when feedback was masked. The perceptual analysis showed that speakers were able to use auditory information to partially compensate for the vocal-tract modification. Furthermore, utterances produced during the masked conditions also improved over a session, demonstrating that the compensatory articulations were learned and available after auditory feedback was removed. PMID- 12558290 TI - Individual talker differences in voice-onset-time. AB - Individual talkers differ in the acoustic properties of their speech, and at least some of these differences are in acoustic properties relevant for phonetic perception. Recent findings from studies of speech perception have shown that listeners can exploit such differences to facilitate both the recognition of talkers' voices and the recognition of words spoken by familiar talkers. These findings motivate the current study, whose aim is to examine individual talker variation in a particular phonetically-relevant acoustic property, voice-onset time (VOT). VOT is a temporal property that robustly specifies voicing in stop consonants. From the broad literature involving VOT, it appears that individual talkers differ from one another in their VOT productions. The current study confirmed this finding for eight talkers producing monosyllabic words beginning with voiceless stop consonants. Moreover, when differences in VOT due to variability in speaking rate across the talkers were factored out using hierarchical linear modeling, individual talkers still differed from one another in VOT, though these differences were attenuated. These findings provide evidence that VOT varies systematically from talker to talker and may therefore be one phonetically-relevant acoustic property underlying listeners' capacity to benefit from talker-specific experience. PMID- 12558291 TI - Spectral models of additive and modulation noise in speech and phonatory excitation signals. AB - The article presents spectral models of additive and modulation noise in speech. The purpose is to learn about the causes of noise in the spectra of normal and disordered voices and to gauge whether the spectral properties of the perturbations of the phonatory excitation signal can be inferred from the spectral properties of the speech signal. The approach to modeling consists of deducing the Fourier series of the perturbed speech, assuming that the Fourier series of the noise and of the clean monocycle-periodic excitation are known. The models explain published data, take into account the effects of supraglottal tremor, demonstrate the modulation distortion owing to vocal tract filtering, establish conditions under which noise cues of different speech signals may be compared, and predict the impossibility of inferring the spectral properties of the frequency modulating noise from the spectral properties of the frequency modulation noise (e.g., phonatory jitter and frequency tremor). The general conclusion is that only phonatory frequency modulation noise is spectrally relevant. Other types of noise in speech are either epiphenomenal, or their spectral effects are masked by the spectral effects of frequency modulation noise. PMID- 12558292 TI - Unfolding of phonetic information over time: a database of Dutch diphone perception. AB - We present the results of a large-scale study on speech perception, assessing the number and type of perceptual hypotheses which listeners entertain about possible phoneme sequences in their language. Dutch listeners were asked to identify gated fragments of all 1179 diphones of Dutch, providing a total of 488,520 phoneme categorizations. The results manifest orderly uptake of acoustic information in the signal. Differences across phonemes in the rate at which fully correct recognition was achieved arose as a result of whether or not potential confusions could occur with other phonemes of the language (long with short vowels, affricates with their initial components, etc.). These data can be used to improve models of how acoustic-phonetic information is mapped onto the mental lexicon during speech comprehension. PMID- 12558293 TI - Discrete-time modeling of woodwind instrument bores using wave variables. AB - A method for simulation of acoustical bores, useful in the context of sound synthesis by physical modeling of woodwind instruments, is presented. As with previously developed methods, such as digital waveguide modeling (DWM) [Smith, Comput. Music J. 16, 74-91 (1992)] and the multi convolution algorithm (MCA) [Martinez et al., J. Acoust. Soc. Am. 84, 1620-1627 (1988)], the approach is based on a one-dimensional model of wave propagation in the bore. Both the DWM method and the MCA explicitly compute the transmission and reflection of wave variables that represent actual traveling pressure waves. The method presented in this report, the wave digital modeling (WDM) method, avoids the typical limitations associated with these methods by using a more general definition of the wave variables. An efficient and spatially modular discrete-time model is constructed from the digital representations of elemental bore units such as cylindrical sections, conical sections, and toneholes. Frequency-dependent phenomena, such as boundary losses, are approximated with digital filters. The stability of a simulation of a complete acoustic bore is investigated empirically. Results of the simulation of a full clarinet show that a very good concordance with classic transmission-line theory is obtained. PMID- 12558294 TI - Suppression of large intraluminal bubble expansion in shock wave lithotripsy without compromising stone comminution: refinement of reflector geometry. AB - Using the Hamilton model [Hamilton, J. Acoust. Soc. Am. 93, 1256-1266 (1993)], the effects of reflector geometry on the pulse profile and sequence of the shock waves produced by the original and upgraded reflector of an HM-3 lithotripter were evaluated qualitatively. Guided by this analysis, we have refined the geometry of the upgraded reflector to enhance its suppressive effect on intraluminal bubble expansion without compromising stone comminution in shock wave lithotripsy. Using the original HM-3 reflector at 20 kV, rupture of a standard vessel phantom made of cellulose hollow fiber (i.d. = 0.2 mm), in which degassed water seeded with ultrasound contrast agents was circulated, was produced at the lithotripter focus after about 30 shocks. In contrast, using the upgraded reflector at 24 kV no rupture of the vessel phantom could be produced within a 20-mm diameter around the lithotripter focus even after 200 shocks. On the other hand, stone comminution was comparable between the two reflector configurations, although slightly larger fragments were produced by the upgraded reflector. After 2000 shocks, stone comminution efficiency produced by the original HM-3 reflector at 20 kV is 97.15 +/- 1.92% (mean +/- SD), compared to 90.35 +/- 1.96% produced by the upgraded reflector at 24 kV (p<0.02). All together, it was found that the upgraded reflector could significantly reduce the propensity for vessel rupture in shock wave lithotripsy while maintaining satisfactory stone comminution. PMID- 12558295 TI - Echolocation signals of wild Atlantic spotted dolphin (Stenella frontalis). AB - An array of four hydrophones arranged in a symmetrical star configuration was used to measure the echolocation signals of the Atlantic spotted dolphin (Stenella frontalis) in the Bahamas. The spacing between the center hydrophone and the other hydrophones was 45.7 cm. A video camera was attached to the array and a video tape recorder was time synchronized with the computer used to digitize the acoustic signals. The echolocation signals had bi-modal frequency spectra with a low-frequency peak between 40 and 50 kHz and a high-frequency peak between 110 and 130 kHz. The low-frequency peak was dominant when the signal the source level was low and the high-frequency peak dominated when the source level was high. Peak-to-peak source levels as high as 210 dB re 1 microPa were measured. The source level varied in amplitude approximately as a function of the one-way transmission loss for signals traveling from the animals to the array. The characteristics of the signals were similar to those of captive Tursiops truncatus, Delphinapterus leucas and Pseudorca crassidens measured in open waters under controlled conditions. PMID- 12558296 TI - Echolocation in the Risso's dolphin, Grampus griseus. AB - The Risso's dolphin (Grampus griseus) is an exclusively cephalopod-consuming delphinid with a distinctive vertical indentation along its forehead. To investigate whether or not the species echolocates, a female Risso's dolphin was trained to discriminate an aluminum cylinder from a nylon sphere (experiment 1) or an aluminum sphere (experiment 2) while wearing eyecups and free swimming in an open-water pen in Kaneohe Bay, Hawaii. The dolphin completed the task with little difficulty despite being blindfolded. Clicks emitted by the dolphin were acquired at average amplitudes of 192.6 dB re 1 microPa, with estimated sources levels up to 216 dB re 1 microPa-1 m. Clicks were acquired with peak frequencies as high as 104.7 kHz (Mf(p) = 47.9 kHz), center frequencies as high as 85.7 kHz (Mf(0) = 56.5 kHz), 3-dB bandwidths up to 94.1 kHz (M(BW) = 39.7 kHz), and root mean-square bandwidths up to 32.8 kHz (M(RMS) = 23.3 kHz). Click durations were between 40 and 70 micros. The data establish that the Risso's dolphin echolocates, and that, aside from slightly lower amplitudes and frequencies, the clicks emitted by the dolphin were similar to those emitted by other echolocating odontocetes. The particular acoustic and behavioral findings in the study are discussed with respect to the possible direction of the sonar transmission beam of the species. PMID- 12558297 TI - The influence of flight speed on the ranging performance of bats using frequency modulated echolocation pulses. AB - Many species of bat use ultrasonic frequency modulated (FM) pulses to measure the distance to objects by timing the emission and reception of each pulse. Echolocation is mainly used in flight. Since the flight speed of bats often exceeds 1% of the speed of sound, Doppler effects will lead to compression of the time between emission and reception as well as an elevation of the echo frequencies, resulting in a distortion of the perceived range. This paper describes the consequences of these Doppler effects on the ranging performance of bats using different pulse designs. The consequences of Doppler effects on ranging performance described in this paper assume bats to have a very accurate ranging resolution, which is feasible with a filterbank receiver. By modeling two receiver types, it was first established that the effects of Doppler compression are virtually independent of the receiver type. Then, used a cross-correlation model was used to investigate the effect of flight speed on Doppler tolerance and range-Doppler coupling separately. This paper further shows how pulse duration, bandwidth, function type, and harmonics influence Doppler tolerance and range Doppler coupling. The influence of each signal parameter is illustrated using calls of several bat species. It is argued that range-Doppler coupling is a significant source of error in bat echolocation, and various strategies bats could employ to deal with this problem, including the use of range rate information are discussed. PMID- 12558298 TI - Measuring hearing in the harbor seal (Phoca vitulina): comparison of behavioral and auditory brainstem response techniques. AB - Auditory brainstem response (ABR) and standard behavioral methods were compared by measuring in-air audiograms for an adult female harbor seal (Phoca vitulina). Behavioral audiograms were obtained using two techniques: the method of constant stimuli and the staircase method. Sensitivity was tested from 0.250 to 30 kHz. The seal showed good sensitivity from 6 to 12 kHz [best sensitivity 8.1 dB (re 20 microPa2 x s) RMS at 8 kHz]. The staircase method yielded thresholds that were lower by 10 dB on average than the method of constant stimuli. ABRs were recorded at 2, 4, 8, 16, and 22 kHz and showed a similar best range (8-16 kHz). ABR thresholds averaged 5.7 dB higher than behavioral thresholds at 2, 4, and 8 kHz. ABRs were at least 7 dB lower at 16 kHz, and approximately 3 dB higher at 22 kHz. The better sensitivity of ABRs at higher frequencies could have reflected differences in the seal's behavior during ABR testing and/or bandwidth characteristics of test stimuli. These results agree with comparisons of ABR and behavioral methods performed in other recent studies and indicate that ABR methods represent a good alternative for estimating hearing range and sensitivity in pinnipeds, particularly when time is a critical factor and animals are untrained. PMID- 12558299 TI - High intensity anthropogenic sound damages fish ears. AB - Marine petroleum exploration involves the repetitive use of high-energy noise sources, air-guns, that produce a short, sharp, low-frequency sound. Despite reports of behavioral responses of fishes and marine mammals to such noise, it is not known whether exposure to air-guns has the potential to damage the ears of aquatic vertebrates. It is shown here that the ears of fish exposed to an operating air-gun sustained extensive damage to their sensory epithelia that was apparent as ablated hair cells. The damage was regionally severe, with no evidence of repair or replacement of damaged sensory cells up to 58 days after air-gun exposure. PMID- 12558300 TI - A comparison of the fragmentation thresholds and inertial cavitation doses of different ultrasound contrast agents. AB - Contrast bubble destruction is important in several new diagnostic and therapeutic applications. The pressure threshold of destruction is determined by the shell material, while the propensity for of the bubbles to undergo inertial cavitation (IC) depends both on the gas and shell properties of the ultrasound contrast agent (UCA). The ultrasonic fragmentation thresholds of three specific UCAs (Optison, Sonazoid, and biSpheres), each with different shell and gas properties, were determined under various acoustic conditions. The acoustic emissions generated by the agents, or their derivatives, characteristic of IC after fragmentation, was also compared, using cumulated broadband-noise emissions (IC "dose"). Albumin-shelled Optison and surfactant-shelled Sonazoid had low fragmentation thresholds (mean = 0.13 and 0.15 MPa at 1.1 MHz, 0.48 and 0.58 MPa at 3.5 MHz, respectively), while polymer-shelled biSpheres had a significant higher threshold (mean = 0.19 and 0.23 MPa at 1.1 MHz, 0.73 and 0.96 MPa for thin and thick-shell biSpheres at 3.5 MHz, respectively, p<0.01). At comparable initial concentrations, surfactant-shelled Sonazoid produced a much larger IC dose after shell destruction than did either biSpheres or Optison (p<0.01). Thick shelled biSpheres had the highest fragmentation threshold and produced the lowest IC dose. More than two and five acoustic cycles, respectively, were necessary for the thin- and thick-shell biSpheres to reach a steady-state fragmentation threshold. PMID- 12558301 TI - Microparticle column geometry in acoustic stationary fields. AB - Particles suspended in a fluid will experience forces from stationary acoustic fields. The magnitude of the force depends on the time-averaged energy density of the field and the material properties of the particles and fluid. Forces acting on known particles smaller than 20 microm were studied. Within a 500 kHz acoustic beam generated by a plane-piston circular source, observations were made of the geometry of the particle column that is formed. Varying the acoustic energy altered the column width in a manner predicted by equations for the primary acoustic radiation force from scattering of particles in the long-wavelength limit. The minimum pressures required to trap gas, solid, and liquid particles in a water medium at room temperature were also estimated to within 12%. These results highlight the ability of stationary acoustic fields from a plane-piston radiator to impose nano-Newton-scale forces onto fluid particles with properties similar to biological cells, and suggest that it is possible to accurately quantify these forces. PMID- 12558302 TI - Who put the tank in the tiger? PMID- 12558303 TI - Cardiocirculatory bio-assist: is it time to reconsider demand dynamic cardiomyoplasty? Review and future perspectives. AB - In the last 15 years, dynamic cardiomyoplasty has remained an experimental procedure even after the enthusiastic short- and mid-term results, mainly because of the disappointing long-term outcome caused by muscular degeneration secondary to chronic continuous electrical stimulation of the latissimus dorsi. In Italy, a group of muscular pathologists, cardiologists, and cardiac surgeons conducted an experiment of an activity-rest stimulation protocol in humans that should avoid complete transformation of the skeletal muscle, maintaining its properties overtime. This "demand" stimulation protocol gave good results, improving New York Heart Association class, ejection fraction value, and survival. Even though dynamic cardiomyoplasty was excluded from the recent international guidelines for the management of heart failure, the discussion on the ability of this unique kind of cardiocirculatory bio-assistance is due to be reopened, thanks to the results of the new stimulation protocol. Heart transplantation, circulatory supporting devices, multisite stimulation therapy, and the total artificial heart are not always and in all countries the best solutions: the great economic cost, the numerous contraindications, the need for immunosuppression and antithrombotic therapy, and the troublesome follow up constitute important drawbacks. For patients in whom transplant surgery cannot be performed, as well as in developing countries, the nonprohibitively expensive demand dynamic cardiomyoplasty may still play a role. PMID- 12558304 TI - The effect of augmented hemodynamics on blood flow during arteriovenous carbon dioxide removal. AB - Arteriovenous carbon dioxide removal (AVCO2R) as an alternative treatment for acute respiratory distress syndrome uses a low resistance gas exchanger in a simple arteriovenous shunt to achieve total CO2 removal and allow lung rest. We have previously shown in our clinically relevant LD40 ovine model of smoke/burn induced acute respiratory distress syndrome that AVCO2R allows significant decreases in respiratory rate, tidal volume, peak airway pressure, and FiO2, as compared with standard mechanical ventilation. In addition, we have shown in a prospective randomized outcomes study that AVCO2R increases ventilator free days, decreases ventilator dependent days, and significantly improves survival. The purpose of this study is to further define the limits of AVCO2R through hemodynamic augmentation and evaluation of peak end expiratory pressure (PEEP). Administration of an alpha agonist (phenylephrine) and a beta agonist (isoproterenol) increased mean arterial pressure (MAP) and cardiac output (CO), respectively. MAP increases ranged from 2.4% to 94.4% and CO increases ranged from 33% to 146%. Phenylephrine caused elevations in MAP (2.4-94.4%) and AVCO2R flow (9-67%), and CO never decreased more than 10%. Isoproterenol administration increased CO (33-146%), decreased MAP (9-54%), and decreased AVCO2R flow (11 42%). In a second group, PEEP was increased stepwise from 0 (baseline) to 20 cm H2O. Increasing PEEP did not result in significant hemodynamic changes (< 10% change from baseline PEEP) for MAP, CO, or AVCO2R flow. In conclusion, alpha agonist administration increased AVCO2R blood flow, whereas beta agonist administration decreased MAP and AVCO2R blood flow, despite CO elevation. Various levels of PEEP are well tolerated and thus allow a range of options during AVCO2R. PMID- 12558305 TI - Design of an artificial lung compliance chamber for pulmonary replacement. AB - Matching the impedance of an artificial lung for pulmonary replacement to native pulmonary impedance is important in preventing right ventricular dysfunction. A lumped-parameter theoretical model and bench-top experiments were used to investigate the effect of a prototype compliance chamber on input impedance. The bench-top simulation consisted of a pulsatile flow generator, a prototype compliance chamber, and a low resistance artificial lung connected in series. Effective compliance was varied using pneumatic compression. The theoretical model considered a similar circuit with resistors before and after a compliance element. The bundle flow pulsatility (flow amplitude divided by average flow) and input impedance were calculated in the theoretical and experimental models. More compliance and lower upstream resistance result in lower bundle flow pulsatility and reduced first harmonic impedance. Matching the time scale of the circuit to the period of pulsatile flow also reduces impedance. The bench-top circuit demonstrated an optimal chamber pressure at which first harmonic impedance is reduced by 80%. The prototype compliance chamber in series with the artificial lung more closely matches native pulmonary impedance. The lumped-parameter model and the bench-top simulation will aid in the design and testing of compliance chamber modifications to improve its efficiency. PMID- 12558306 TI - Plasma concentrations of midazolam in neonates receiving extracorporeal membrane oxygenation. AB - Drug disposition is affected during extracorporeal membrane oxygenation (ECMO). This study investigates the dose-concentration relationship of midazolam in neonates requiring ECMO during continuous infusion into the circuit (extracorporeally; n = 10) and intravenously (n = 10). Data on hourly doses and sedation scores were collected for 120 hours. Plasma concentrations were analyzed at times 0, 2, 4, 6, 12, 18, and 24, and every 12 hours thereafter. Both groups were clinically similar. Mean (standard deviation) dose for all patients was 250 (185) microg/kg/h, four times greater than previously reported. Doses administered in the first 24 hours were significantly greater extracorporeally [361 (300)] compared with intravenous [258 (190) microg/kg/h, p < 0.001]. Mean (standard deviation) plasma concentrations in all patients at 24, 48, and 72 hours were 1.4 (0.9), 1.8 (1.2), and 2.6 (1.8) microg/ml, respectively. Satisfactory sedation levels were achieved in all patients. Comparison of the actual observed with predicted (simulated) midazolam concentrations suggested significant attenuation of plasma levels during the first 24 hours of ECMO. However, at 48 hours, observed concentrations exceeded those predicted, suggesting accumulation. We conclude that in the first 24 hours of ECMO, because of an expanded circulating volume and sequestration by the circuit, significantly more midazolam is required to achieve adequate sedation. Subsequently, and because of circuit saturation, maintenance doses should be reduced. PMID- 12558307 TI - Transplantation of pig metanephroi. AB - To determine whether pig metanephroi grow and differentiate after allotransplantation or xenotransplantation across a highly disparate barrier, we implanted metanephroi from embryonic day 28 (E28) pig embryos into the omentum of unilaterally nephrectomized adult pigs or C57Bl/6J mice (hosts). Some mouse hosts received anti-CD45RB, anti-CD154, and anti-CD11a (costimulatory blockade). E28 pig metanephroi were < 0.2 mm in diameter and contained only metanephric blastema and segments of ureteric bud. Pig metanephroi transplanted into pigs underwent growth and differentiation of nephrons over a 2 week period without the need for costimulatory blockade of hosts. In contrast, pig metanephroi did not grow or differentiate in mice that received no costimulatory blockade. However, by 2 weeks posttransplantation in mice in which costimulation was blocked, metanephroi from E28 pigs had enlarged, become vascularized, and had formed mature tubules and glomeruli. By 3 weeks posttransplantation in mice, metanephroi had grown to the point that they were approximately half the volume of the native mouse kidney. Here we show that growth and development of pig metanephroi occurs posttransplantation across an allogeneic or highly disparate xenogeneic barrier. PMID- 12558308 TI - Neoorganogenesis and neohistogenesis by desired metaplasia of autogenous tissue stem cells in vivo: a critical and scientific evaluation with 125 years of review literature. AB - Organogenesis and histogenesis during embryogenesis is well known. In the adult, it is minimal or absent in higher animals-mammals and humans. Neoformation of tissues and organs in vivo in dogs, monkey, and humans has been demonstrated using autogenous pluripotent stem cells. This is possible with the body's own potential, and the phenomena has been described as "desired metaplasia." The stem cells are surgically colonized with the tissues and/or tissue system where the tissue or organ is to be regenerated. Neoformation of abdominal wall aponeurosis, ureter, etc. has been possible by this technique. The technology has also been used in the management of human diseases like large incisional hernias and complex genitourinary or rectal fistulae. Because desired metaplasia is a new concept, the literature of the past 125 years has been scrutinized, selected articles on experimental colonization of tissues and cells have been analytically studied, and relevant articles have been critically analyzed with a new found concept of desired metaplasia and the proposed hypothesis of the first author based on embryologic principles. The literature reviewed revealed no article with the concept of desired metaplasia being responsible for neoorganogenesis and neohistogenesis. New organ and tissue formation is possible with the help of autogenous pluripotent stem cells if exposed to the proper environment and functional need, provided the tissues are embryologic neighbors. The phenomenon of desired metaplasia is discussed on a scientific basis. PMID- 12558309 TI - Cartilage regeneration using slow release of bone morphogenetic protein-2 from a gelatin sponge to treat experimental canine tracheomalacia: a preliminary report. AB - We investigated whether saber sheath-type tracheomalacia could be treated by the slow release of bone morphogenetic protein (BMP)-2 from a gelatin sponge. A 1 cm gap was made in the middle portion of each of 10 consecutive tracheal cartilage rings in the canine cervix (control group, n = 3), then a gelatin sponge containing 12 microg of BMP-2 solution was implanted in the gap (12 microg group, n = 3). In another group (120 microg + P group, n = 3), the implanted gelatin sponge contained 120 microg of BMP-2 solution, and the gap was covered with periosteum. All of the control dogs developed saber sheath-type tracheomalacia, whereas tracheomalacia was not observed in the 12 microg and 120 microg + P groups. In the 12 microg group, fibrous cartilage was observed at the ends of the cartilage stumps. In the 120 microg + P group, newly formed bone and cartilage were observed to form a bridge between the cartilage stumps. The regeneration of cartilage or bone induced by the slow release of BMP-2 from a gelatin sponge might be useful for treatment of tracheomalacia. PMID- 12558310 TI - Biochemical effects of high dialysate calcium in hemodialysis patients with hyperparathyroidism: a 10 month study. AB - In the past 15 years, there has been a trend to decrease dialysate calcium concentrations to prevent hypercalcemia. However, low dialysate calcium can provoke hyperparathyroidism. The time course of the effect of increasing dialysate calcium is not well characterized, and the effect on calcium-phosphate product is unclear. Therefore, we studied the effect of increasing dialysate calcium from 1.5 to 1.75 mM in 21 stable patients on hemodialysis who had serum phosphate of less than 2 mM and serum calcium of less than 2.4 mM. Over 10 months, parathyroid hormone levels fell from 39.6 to 16.6 pM (p < 0.0001), whereas serum calcium increased from 2.27 to 2.41 mM. There were no significant changes in serum phosphate or the calcium-phosphate product. Three patients became hypercalcemic when their parathyroid hormone levels were suppressed to less than 10 pM. We conclude that in carefully selected patients, increasing dialysate calcium can safely treat hyperparathyroidism with minimal risk of complications. This treatment has the advantage over the use of vitamin D therapy of being less expensive, independent of patient compliance, and less likely to cause increases in serum phosphate or calcium-phosphate product. PMID- 12558311 TI - Neuropeptide-Y and atrial natriuretic peptide as prognostic markers in patients on hemodialysis. AB - We conducted a study of the influence of the vasoactive peptides atrial natriuretic peptide (ANP) and neuropeptide Y (NPY) on survival of patients on hemodialysis and their association and relative importance with cardiac and clinical variables. Thirty-three hemodialysis patients were characterized by age, sex, diagnosis, blood pressure, serum (S)-albumin, serum (S)-urea, hemoglobin, dialysis dose, weight gain, duration of dialysis, cardiac hypertrophy, volume, failure, and ischemia and plasma levels of ANP and NPY. The outcomes were analyzed for early deaths (< 1 year) and for all deaths. The association of the variables to early deaths and all deaths, respectively, was studied in Cox proportional hazard analyses. The variables were also studied in three hierarchical steps: clinical variables only, clinical and cardiac variables, and all variables. For all deaths, the independent variables were plasma NPY (pmol/L) (hazard ratio [HR] = 1.035, p = 0.004), heart volume (ml/m2) (HR = 1.009, p = 0.001), and S-albumin (g/L) (HR = 0.750, p = 0.034). For early deaths, the independent variables were predialysis ANP (pmol/L) (HR = 1.008, p = 0.034) and NPY (pmol/L) (HR = 1.031, p = 0.026). In the hierarchical study, excluding the vasoactive peptides, heart volume, heart failure and S-albumin were independently associated with all deaths, and mean arterial blood pressure was associated with early death. When also excluding the cardiac parameters, S-albumin was associated with all deaths and mean arterial blood pressure with early death. In conclusion, plasma levels of the vasoactive peptides ANP and NPY are the most important group in a hierarchy of variables that predict imminent death in hemodialysis patients, and NPY is associated with late death. ANP and NPY apparently sum up the detrimental influence of many factors in hemodialysis patients. PMID- 12558312 TI - Hollow fiber shape alters solute clearances in high flux hemodialyzers. AB - The mass transfer properties of hemodialyzers containing hollow fiber membranes are known to be influenced by membrane chemical composition, surface area, and pore size; however, the effects of hollow fiber shape (or configuration) and packing density within the dialyzer housing have not been well characterized. We determined, both in vitro and ex vivo (clinical), solute clearances and mass transfer-area coefficients (KoA) for high flux dialyzers containing polysulfone hollow fibers of identical chemical composition but different shapes. Hemoflow F80A (1.8 m2 of membrane surface area) dialyzers contained hollow fibers with a conventional shape, but Optiflux F180A (1.8 m2), F200A (2.0 m2), and F200NR (2.0 m2) dialyzers contained hollow fibers with a wavy shape. Clearances and KoA values determined in vitro for urea and creatinine increased with increasing dialysate flow rate and were higher for Optiflux F180A and F200A dialyzers than for Hemoflow F80A dialyzers. In vitro clearances for lysozyme and myoglobin were also higher for Optiflux F180A and F200A dialyzers than for Hemoflow F80A dialyzers, suggesting that a wavy hollow fiber shape increases mass transfer by increasing effective membrane surface area, conceivably by altering dialysate flow patterns. Urea clearances and KoA values determined ex vivo were higher for Optiflux F200NR dialyzers than for Hemoflow F80A dialyzers, confirming that the in vitro results are applicable to clinical hemodialysis. These increases in mass transfer efficiency for dialyzers containing hollow fibers with a wavy shape are consistent with improved mass transfer within the dialysate compartment as evidenced by the manufacturer-reported dialysate pressure-flow relationships. We conclude that the mass transfer characteristics of high flux dialyzers can be altered by the shape of the hollow fibers. PMID- 12558313 TI - Sympathetic skin response in peritoneal dialysis patients. AB - To determine if there is any difference in nerve conduction studies or sympathetic skin response (SSR) between patients on peritoneal dialysis and those on regular hemodialysis, we did a cross-sectional observational study. The study group consisted of 24 patients on peritoneal dialysis (PD) (12 men, aged 45 +/- 17 years) and 20 patients on hemodialysis (HD) (11 men, aged 50 +/- 22 years). All of these patients were in stable clinical condition, they were receiving adequate dialysis, and none of them had systemic diseases. Motor and sensory nerve conduction studies of the common and medial peroneal nerve and SSR were performed in all patients. There were no differences in motor and sensory nerve conduction velocities between PD and HD patients. All PD patients had detectable SSR. However, six patients on HD (30%) failed to show SSR (p < 0.05). Mean SSR amplitude was higher in PD patients than in HD patients (1233 +/- 843 vs. 605 +/- 771 microv, p < 0.05). There were no differences in mean SSR latency between PD and HD patients. PD modality (continuous ambulatory PD vs. automated PD) or the presence of residual renal function did not influence nerve conduction studies or SSR. In conclusion, using standard nerve conduction studies, no differences could be found between HD and PD. However, a higher proportion of patients on HD showed an impaired SSR, suggesting that subclinical neuropathy may be more common in HD than PD patients. PMID- 12558314 TI - Retrospective evaluation of renal kt/V(urea) at the initiation of long-term peritoneal dialysis at the University of Missouri: relationships to longitudinal nutritional status on peritoneal dialysis. AB - The purpose of this study was to examine the impact of low levels of residual renal function (RRF) on nutritional status in end-stage renal disease patients starting peritoneal dialysis (PD) at baseline and after a year on dialysis. We conducted a single center retrospective analysis of 116 patients who started long term PD in a university teaching hospital from 1989 to 1998 and were followed for 1 year. Patients were divided into four equal groups according to their initial renal Kt/V(urea) (L/week) levels at the start of PD and followed for 1 year. There were no interventions. The relationship between dialysis adequacy (renal and total Kt/V(urea)) and nutritional status was studied at baseline and at 1 year. Baseline data for patients who survived were compared with the baseline data of those who died and with their own 1 year data. At baseline, the mean serum albumin (3.31 g/dl, p < 0.0001) and lean body mass (47.20% body weight, p < 0.04) of group 1 were significantly lower than in groups 2, 3, and 4. Levels of normalized protein equivalent of nitrogen appearance (nPNA) were significantly lower in group 1 than in groups 3 and 4 (p < 0.005). Although group 1 patients showed trends toward improvement in nutritional parameters, they never caught up with the other groups. At the end of 1 year, the lower total Kt/V(urea) in group 1, with the lowest RRF, was associated with the lowest mean values for nutritional status and the highest death rate. Comparison of baseline and 1 year data of survivors showed that nutritional status improved or remained stable in groups 3 and 4, who exceeded the minimum recommended adequacy targets as per Dialysis Outcome Quality Initiative criteria (mean 12 month total Kt/V(urea) 2.18 and 2.58, respectively). Comparison of baseline data of survivors and those who died showed that patients who died had lower mean values for serum albumin, nPNA, lean body mass, and body weight across all groups. Low RRF at the start of dialysis is associated with poor nutritional status. Also, patients who start dialysis with low RRF and poor nutritional status do not have significantly improved nutritional status even after 1 year on dialysis. PMID- 12558315 TI - Effects of continuous flow left ventricular assist device support on skin tissue microcirculation and aortic hemodynamics. AB - Continuous flow ventricular assist devices (CFVADs) are thought to be the next generation of circulatory assist devices. With many now in various stages of development or clinical trial, it is important that the physiologic aspects of these pumps be critically analyzed. In this study, 15 calves were divided into two groups. One group received a CFVAD, and the other a sham implant. Two additional animals were used in an acute study to examine aortic blood flow patterns from a CFVAD. Tissue perfusion was measured on all animals before surgery and then weekly thereafter. Before surgery, there was no difference in hemodynamics or tissue perfusion between studied animals. Postoperatively, CFVAD animals had statistically significant increased diastolic pressure. Significantly decreased pulse pressure, pulse index, and tissue perfusion were also observed in CFVAD animals. Results from the flow pattern studies suggested that at moderate levels of pump support (40-75%), the amount of blood flow distal to the outflow graft anastomosis decreased approximately 25% because of increased regurgitant blood flow in the aorta. These results suggest that the diminished tissue perfusion is likely due to changes in aortic hemodynamics and provide some insight into the distribution of flow from CFVADs. PMID- 12558316 TI - Usefulness of continuous air tonometry for evaluation of splanchnic perfusion during cardiopulmonary bypass. AB - Although gastric mucosal tonometry has been reported as a useful method to assess splanchnic perfusion during cardiovascular surgery, the conventional discontinuous method of tonometry (saline tonometry) was cumbersome and prone to systematic errors. A new automated system of air tonometry (Tonocap; Datex Ohmeda, Helsinki, Finland) allows for frequent (every 10 minutes) measurement of gastric regional CO2 (PrCO2) and may be more suitable as a monitoring system in cardiac patients. We evaluated the usefulness of continuous air tonometry as a marker of splanchnic perfusion during cardiopulmonary bypass (CPB). In 19 patients (53-79 years, mean 63 years) who underwent cardiovascular surgery under standard CPB with mild hypothermia (32 degrees C) from January 2001 to May 2002, the PrCO2 and calculated intramucosal pH (pHi) of gastric tonometry was monitored using Tonocap, and their relation to postoperative visceral organ function was evaluated. The pHi significantly increased after initiation of CPB from 7.32 +/- 0.07 to 7.43 +/- 0.10 (p < 0.05) and then consistently decreased in all patients to 7.39 +/- 0.09 at the end of CPB. The value of PrCO2 significantly (p < 0.01) correlated with the value of pHi. The lowest value of pHi during CPB was significantly related to blood urea nitrogen (r = -0.75, p < 0.05), serum creatinine (r = -0.78, p < 0.05), creatinine clearance (r = 0.68, p < 0.05) on postoperative day 1, and blood urea nitrogen (r = -0.84, p < 0.01) on day 3. In contrast, arterial blood lactate level, venous oxygen saturation, and routinely measured hemodynamics (e.g., pump flow, arterial pressure) during CPB were unrelated to the postoperative visceral organ function. These results suggest that continuous monitoring of gastric regional CO2 and pHi by air tonometry system is useful for the evaluation of splanchnic perfusion during CPB and may contribute to improve CPB technique by allowing the early detection of visceral malperfusion. PMID- 12558317 TI - Long-term ex vivo bovine experiments with the Gyro C1E3 centrifugal blood pump. AB - Centrifugal blood pumps are used widely for cardiopulmonary bypass, as ventricular assist devices, and for extracorporeal membrane oxygenation (ECMO). However, there is no centrifugal blood pump that is suitable for long-term ECMO. The authors developed the Gyro C1E3 centrifugal blood pump (Kyocera Corporation, Kyoto, Japan), which has superior antithrombogenic, antitraumatic, and hydraulic features in comparison with the conventional centrifugal blood pumps. Five ex vivo long-term durability tests of the Gyro C1E3 were performed using healthy miniature calves. The ECMO circuit was composed of a prototype hollow fiber silicone membrane oxygenator and a Gyro C1E3 pump. Venous blood was drained from the left jugular vein of a calf, passed through the oxygenator and infused into the left carotid artery using a Gyro C1E3. Ex vivo studies were performed from 7 to 15 days at a blood flow rate of 1 L/min. During this period, the Gyro C1E3 demonstrated a stable performance without exchanging the pump. Bleeding complications were the major reason for termination of each experiment. Rotational speed was maintained around 2,000 rpm. All five calves demonstrated neither abnormal signs nor abnormal blood examination data throughout the experiment. Neither clot nor thrombus formations were found during the necropsy in the cannula or pump nor were infarctions observed in any of the major organs. In conclusion, the Gyro C1E3 showed a stable and reliable performance during long term ex vivo bovine experiments under the conditions tested. PMID- 12558318 TI - Blood compatibility of a newly developed trileaflet mechanical heart valve. AB - An ideal heart valve prosthesis, which has both the flow dynamic properties and blood compatibility of a tissue valve prosthesis and the durability of a mechanical prosthesis, does not exist. The Triflo trileaflet mechanical heart valve (MHV; Triflo Medical Inc., Irvine, CA) is a newly developed MHV prosthesis with the following design goals: central flow, minimal flow disturbance and stasis around the hinge region, and durability. The current study was conducted to evaluate the blood compatibility of a 29 mm Triflo MHV in the mitral position of eight calves for 5 months without any postoperative anticoagulation. Whole blood platelet aggregometry and the Xylum Clot Signature Analyzer (Xylum Corporation, Scarsdale, NY) were used to evaluate the postoperative changes in platelet and coagulation functions. Full autopsies, histological examinations of major internal organs, and scanning electron microscopy analyses of the explants were performed. Early termination occurred in two cases; one was because of valve thrombosis on the 25th day, and the other was killed because of a nonvalvular complication on the 105th day. The valve thrombosis was attributed to prolonged ventricular fibrillation at the time of valve replacement surgery. Whole blood platelet aggregometry and clot signature analyzer parameters did not show any sign of activation of platelets or the coagulation system. No hemolysis was observed. There was no macroscopic valve thrombosis or embolism observed in the remaining seven cases. Scanning electron microscopy analyses showed clean leaflet and valve ring surfaces, with only occasional minute platelet aggregations. Excellent blood compatibility of the Triflo MHV was demonstrated in this study. PMID- 12558319 TI - Third model of the undulation pump total artificial heart. AB - The undulation pump is a small, continuous flow displacement type blood pump, and the undulation pump total artificial heart (UPTAH) is a unique, implantable total artificial heart based on this pump. To improve the durability of the UPTAH for investigating long-term pathophysiology with UPTAH, a third model (UPTAH3) has been developed. UPTAH3 was designed to separate the left and right undulation shafts and to be more durable. The undulation pumps were also redesigned. UPTAH3 was implemented with a diameter of 76 mm, width of 78 or 79 mm, total volume of 292 ml, and weight of 620 g. The priming volumes of the left and right pumps are 26 and 21 ml, respectively. The atrial cuffs and outflow cannulae were also redesigned for UPTAH3. The maximum output against an arterial pressure load of 100 mm Hg is about 11 L/min. The maximum pump efficiency is about 15% in the left pump and 18% in the right pump, giving a maximum total efficiency for both of about 11%. To date, UPTAH3 has been tested in 17 goats, and the longest survival period was 46 days. This third model will be useful for investigating pathophysiology with UPTAH. PMID- 12558320 TI - Hemolysis in needleless connectors for phlebotomy. AB - Needleless connectors have been developed recently as a means of reducing transmission of AIDS and other blood-borne diseases by accidental needle sticks. However, the potential for hemolysis induced by fluid stresses within the connector remained to be determined. The influence of needleless connectors on hemolysis was evaluated in simulated clinical blood draws. Blood from five volunteers was drawn with vacuum tubes and syringes through 16, 18, and 22 gauge needles and PosiFlow (Becton Dickinson, Franklin Lakes, NJ) and Clave (ICU Medical, San Clemente, CA) needleless connectors. Hemolysis was measured in the samples by a spectrophotometric technique. Results showed that hemolysis increased when needleless connectors were used. The PosiFlow connector produced more hemolysis than the Clave device. Curiously, erythrocyte damage was greatest for connectors used with 18 gauge needles. Hemolysis was larger for samples drawn with vacuum tubes than with syringes. However, no combinations of connector, needle size, or blood draw device resulted in mean hemolysis values large enough to interfere with clinical assays. PMID- 12558321 TI - A case of erythropoietin induced hypertension in a bilaterally nephrectomized patient. AB - Recombinant human erythropoietin (r-HuEPO) is an established tool for correction of renal anemia. It is well known that chronic administration of r-HuEPO often causes hypertension in dialysis patients. However, the mechanism of the r-HuEPO induced hypertension has not been fully elucidated. We report a case of r-HuEPO induced hypertension in an anephric patient. In this case, hemodialysis was started after removal of both kidneys because of rupture of an angiomyolipoma. Although mean blood pressure (BP) did not change during the period of rapid correction of renal anemia by blood transfusion, treatment with r-HuEPO significantly increased mean BP. Also, discontinuation of r-HuEPO resulted in a decrease in mean BP. These results suggested that r-HuEPO caused an elevation in BP in the absence of kidneys, and the elevation in BP was thought to be independent of an increase in hematocrit level or hypervolemia. We also investigated the role of nitric oxide (NO) in r-HuEPO induced alteration of BP. A significant negative correlation was found between a decrease in serum NO level and an increase in mean BP during the period of r-HuEPO administration. The results suggest that an inhibitory effect of r-HuEPO on NO production might be, at least in part, related to the r-HuEPO induced hypertension in this case. PMID- 12558323 TI - Fundamentals of pulsatile versus nonpulsatile flow during chronic support. PMID- 12558322 TI - First use of the Cancion cardiac recovery system in a human. AB - Animal studies have shown clearly that a peripheral circulatory support system can effectively unload the left ventricle. We report here the first implantation of one such system (Cancion, Orqis Medical, Lake Forest, CA) in a human. The Cancion system consists of a centrifugal pump connected to the circulation via a graft cannula anastomosed to the left axillary artery and a percutaneous cannula placed into the left common femoral artery. Flow is initiated from the femoral to the axillary artery. The system was implanted in a 62-year-old man with ischemic cardiomyopathy suffering from decompensation whose condition had not improved with catecholamine therapy. The Cancion system supported the patient for 3 days, after which it was electively explanted. During the support period, pulmonary capillary wedge pressure dramatically decreased from 28 to 9 mm Hg, left ventricular diastolic dimension decreased from 6.78 to 6.16 cm, creatinine levels decreased from 1.9 to 0.9 mg/dl, cardiac index improved from 1.5 to 2.7 L/min, and ejection fraction improved from 25 to 35%. Together, our data indicate that the Cancion system quickly and effectively improved the patient's hemodynamics. This suggests that the device may one day become a short-term alternative to high dose inotrope therapy and that its application may delay the need for more invasive forms of mechanical circulatory support. PMID- 12558324 TI - Testing a symptom-based surveillance system at high-profile gatherings as a preparatory measure for bioterrorism. AB - We tested symptom-based surveillance during the G8 conference in 2000 as a means of detecting outbreaks, including bio-terrorism attacks, promptly. Five categories of symptoms (skin and haemorrhagic, respiratory, gastrointestinal, neurological and unexplained) were adopted for the case definition of the surveillance. The surveillance began I week before the conference, and continued until 1 week after the conference ended. We could not detect any outbreaks during this surveillance. Compared to the existing diagnosis-based surveillance system, symptom-based surveillance has the advantages of timeliness and simplicity. However, poor specificity and difficulties in determining epidemic threshold were important limitations of this system. To increase the specificity of surveillance, it is essential to incorporate rapid laboratory diagnoses into the system. PMID- 12558325 TI - High resolution genotyping of Campylobacter jejuni strains by macrorestriction analysis with XhoI and polymerase chain reaction targeting enterobacterial repetitive intergenic consensus sequences: can we predict the zoonotic potential of strains? AB - Campylobacter jejuni isolates of human, canine, feline, bovine and poultry origin were investigated for their genomic diversity using O-antigen typing (n = 271), SmaI (n = 158) and XhoI (n = 158) macrorestriction analysis and ERIC-PCR (n = 107). The O-antigens O:1/44, O:2, O:4 complex, O:37. O:40 were identified and 53.7% of the human and 56.1% of the animal strains were typable with the available antisera. Two ERIC-PCR pattern groups were generated representing human and animal strains as well as those exclusively of animal origin. XhoI macrorestriction analysis also distinguished 'human' and 'non-human' strain clusters, but by SmaI restriction mainly serotype-associated clusters were found. In conclusion, genomic differences may occur between 'human' and 'non-human' strains and this may reflect their potential to overcome the barrier from animals to humans. PMID- 12558326 TI - Use of AFLP, plasmid typing and phenotyping in a comparative study to assess genetic diversity of Shigella flexneri strains. AB - Shigella flexneri infections are one of the main causes of acute diarrhoea in Cuba. Twenty strains isolated from sporadic cases in nine different Cuban provinces were characterized. Serotyping, antibiotic-resistance typing, plasmid typing and AFLP-typing were used to determine their suitability for use in epidemiological studies of S. flexneri. The predominant serotypes were serotype 6 (35%) and serotype 2 (35%). Eleven different plasmid profiles were detected (Diversity Index = 0.92). AFLP-typing discriminated 12 different patterns (DI = 0.95), these patterns were not coincident with plasmid-typing patterns. Both techniques combined distinguished 14 patterns among the 20 studied strains (DI = 0.99). There was no consistent relationship between plasmid-typing and AFLP typing patterns or antibiotic-resistance typing patterns. Ninety-five percent of S. flexneri strains were multiresistant. PMID- 12558327 TI - E. coli O157 phage type 21/28 outbreak in North Cumbria associated with pasteurized milk. AB - In March 1999, a large community outbreak of Escherichia coli O157 infection occurred in North Cumbria. A total of 114 individuals were reported to the Outbreak Control Team (OCT); 88 had laboratory confirmed E. coli O157. Twenty eight (32%) of the confirmed cases were admitted to hospital, including three children (3.4%) with haemolytic uraemic syndrome. There were no deaths. A case control study found that illness was strongly associated with drinking pasteurized milk from a local farm (P = <0.0001) on single variable analysis. Microbiological investigations at the farm revealed E. coli O157 phage type (PT) 21/28 VT 2 which was indistinguishable from the human isolates by pulsed field gel electrophoresis. At the time of occurrence this was the largest E. coli O157 outbreak in England and Wales and the first E. coli O157 PT 21/28 VT 2 outbreak associated with pasteurized milk. This outbreak highlights lessons to be learnt regarding on-farm pasteurization. PMID- 12558328 TI - National enhanced surveillance of meningococcal disease in England, Wales and Northern Ireland, January 1999-June 2001. AB - Enhanced surveillance of meningococcal disease (ESMD) was implemented nationally across ten regions of England, Wales and Northern Ireland from 1 January 1999. It aims to deliver more sensitive surveillance than laboratory reporting by including clinically diagnosed but laboratory unconfirmed cases. Consultants in Communicable Disease Control (CsCDC) report all clinically diagnosed cases of meningococcal disease (MD) to the Regional Epidemiologist in the relevant regional unit of the Public Health Laboratory Service (PHLS) Communicable Disease Surveillance Centre (CDSC). These reports are reconciled with laboratory data from the PHLS Meningococcal Reference Unit and then forwarded to the national CDSC where further reconciliation with laboratory data takes place. In addition, CsCDC are asked to report any clusters of MD that occur. Between 1 January 1999 and 30 June 2001, 12,074 cases of MD were ascertained through ESMD. The majority (57%) were laboratory confirmed. The estimated incidence of MD fell between 1999 and 2001 from 9.2 to 8.0 per 100,000 population. Of laboratory confirmed cases, the number of cases of serogroups B and W135 increased and of serogroup C and of ungrouped meningococcal infection decreased. Variation between regions was considerable and deserves further investigation. Of 11,522 cases with a reported clinical diagnosis, 53.6% were diagnosed as septicaemia, 32.6% as meningitis, 12.5% as both septicaemia and meningitis, and 13% had other invasive MD. Between 1 January 1999 and 30 June 2001 698 deaths were reported, an overall case fatality rate (CFR) of 5.8%; 567 deaths were in confirmed cases and 131 probable (CFR 8.2% and 2.5%, respectively). CFR was higher in serogroup C (13.5%) than B (5.8%). No peak in serogroup C meningococcal infection occurred in the winter of 2000/1 and no clusters of serogroup C meningococcal infection were reported in the first half of 2001. ESMD provides information about the epidemiology of MD that is more complete than statutory notification and laboratory surveillance and is useful for evaluating the impact of the meningococcal serogroup C vaccination programme and of the other non-vaccine preventable serogroups. PMID- 12558329 TI - Invasive group A streptococcal infections in the San Francisco Bay area, 1989-99. AB - To describe the epidemiology of invasive group A streptococcal (iGAS) infections in the San Francisco Bay Area, population-based active surveillance for laboratory-confirmed iGAS was conducted by the California Emerging Infections Program in three California counties. From January 1989 to December 1999, 1415 cases of iGAS were identified. Mean iGAS incidence was 4.06/100,000 person-years and case fatality ratio was 13%, with no linear trends over time. Incidence was lowest in adolescents, was higher in men than women (4.4 vs. 3.2/100,000 person years), and was higher in African-Americans (6.7) than in non-Hispanic (4.1) or Hispanic (3.4) Whites, Asians (2.2) or Native Americans (17/100,000 person years). Injecting drug use was the riskiest underlying condition and was associated with the highest attributable risk. Cases were associated with several underlying conditions, but 23% occurred in previously healthy persons. From 1989 1999, iGAS infections in the San Francisco Bay Area became neither more common nor more deadly. PMID- 12558330 TI - Kinetics of the IgG antibody response to pertussis toxin after infection with B. pertussis. AB - We aimed to provide a quantitative description of decay in pertussis antibody levels to aid in finding a serological estimate of the incidence of pertussis. The serum IgG response against pertussis toxin was studied in a group of clinically diagnosed patients. Individual records consisted of repeated serum IgG measurements at irregular intervals for up to 10 years post diagnosis. These data were analysed with a nonlinear regression model taking into account censoring at upper and lower threshold levels, measurement errors, and individual variation in the shape and magnitude of the immune response. There was considerable variation between individual responses, both in strength (amplitude) and duration (shape). The inverse model relating IgG levels to time from infection (diagnosis) can be applied to cross-sectional IgG data to generate distributions of times from infection, which may be used to calculate infection rates and their variation, in populations sampled for cross-sectional IgG data. PMID- 12558331 TI - Nasopharyngeal carriage of resistant pneumococci in young South Indian infants. AB - Streptococcus pneumoniae is the leading bacterial cause of life-threatening infections in infants. Although antibiotic resistance affects management of pneumococcal infections, few data on patterns of resistance are available for India. We examined nasopharyngeal carriage of antibiotic-resistant pneumococci in 464 South Indian infants between 2 and 6 months. Newly acquired serotypes were screened for susceptibility to cotrimoxazole, erythromycin and penicillin using disk diffusion. Cumulative prevalence of pneumococcal carriage rose from 53.9% at 2 months to 70.2% at 6 months. The prevalence of strains that were not susceptible to penicillin, cotrimoxazole and erythromycin was 34, 81.1 and 37.2%, respectively. Carriage of erythromycin non-susceptible strains declined significantly between ages 4 months and 6 months (44.1 vs. 10.7%). More than 87% of the isolates screened were non-susceptible to > or = 1 antibiotic. Serogroups/types that were most frequently non-susceptible to 1 or more antibiotics were 6, 9, 14, 19 and 23. Less than 1% of the isolates were multi drug resistant. Widespread use of antibiotics in South India has resulted in S. pneumoniae becoming non-susceptible to some commonly used antibiotics. Monitoring trends in antibiotic susceptibility and making antibiotics available only through prescription from a health care worker may slow the spread of resistant pneumococci and improve management of pneumococcal infections in South India. PMID- 12558332 TI - Is household antibiotic use a risk factor for antibiotic-resistant pneumococcal infection? AB - We used microbiology and pharmacy data from health-maintenance organizations to determine whether antibiotic use by a household member increases the risk of penicillin-non-susceptible pneumococcal disease. Though it has been well established that an individual's antibiotic use increases one's risk of antibiotic-resistant infection, it is unclear whether the risk is increased if a member of one's household is exposed to antibiotics. We therefore conducted a case-control study of patients enrolled in health maintenance organizations in Western Washington and Northern California. Cases were defined as individuals with penicillin-non-susceptible pneumococcal infection; controls were individuals with penicillin-susceptible pneumococcal infection. Socioeconomic variables were obtained by linking addresses with 1997 census block group data. One-hundred and thirty-four cases were compared with 798 controls. Individual antibiotic use prior to diagnosis increased the odds of penicillin non-susceptibility, with the strongest effect seen for beta-lactam use within 2 months (OR 1.8, 95% CI 1.2, 2.8). When household antibiotic use by persons other than the patient were considered, at 4 months prior to diagnosis there was a trend towards an association between penicillin non-susceptibility and beta-lactam antibiotic use, and a possible association in a small subgroup of patients with eye and ear isolates. However, no significant overall pattern of association was seen. We conclude that though antibiotic use of any kind within 2 months prior to diagnosis is associated with an increased risk of penicillin-non-susceptible pneumococcal disease, there is no significant overall pattern of association between household antibiotic use and penicillin-non-susceptible pneumococcal infection. PMID- 12558333 TI - Antibiotic use, gastroenteritis and respiratory illness in South Australian children. AB - This study examines the incidence of antibiotic-associated diarrhoea (AAD) in children in the community setting. Parents of 965 children aged 4-6 years and resident in rural/semi-rural South Australia completed a questionnaire on socio demographic factors, and a 6-week daily diary detailing symptoms of gastroenteritis, antibiotic use, respiratory illness, and contact with someone with gastroenteritis. The incidence of AAD was 32.3 %, falling to 23.5% when episodes associated with a respiratory illness were excluded. Respiratory illness in the previous 3 days (OR 6.76, 95% CI 4.87, 9.38), and contact with someone with gastroenteritis in the previous 14 days (OR 1.8, 95% CI 1.48, 2.19), were both associated with gastroenteritis. After adjusting for these, only the first day of antibiotic use was associated with gastroenteritis (OR 3.8, 95% CI 1.8, 8.06). Potential confounding factors, in particular inter-current respiratory illness, need to be considered when examining AAD. PMID- 12558334 TI - Comparison between cohorts vaccinated and unvaccinated against influenza and pneumococcal infection. AB - This study characterizes possible confounders that might make cohorts vaccinated and unvaccinated against influenza and pneumococcal infection different at baseline, with the hypothesis that the two cohorts are comparable. The similarity between health and demographic data was analysed by a randomized, multivariant study addressed to 10,000 persons aged 65 years and older in Stockholm County and was carried out in the form of a postal inquiry during the period December 2000 to May 2001. The study-population response rate was 78%. Of these, 66% (5,120 persons) had been given at least one influenza vaccination during the 3-year study period (1998-2000), 50% (3,780) had received one pneumococcal vaccination and 78% had received both vaccines during the period. The vaccination rate was lower in the age group 65-69 years (60%), compared with elderly cohorts aged over 70 years (67-72%, P < 0.001). Elderly persons living in nursing homes or institutions had higher vaccination rates than persons living in their own households (72 vs. 67%). Persons with underlying chronic diseases had higher vaccination rates (71%, P < 0.001) than those without underlying chronic diseases. Vaccine recipients had fewer days in hospital, compared with non recipients. Unvaccinated persons with myocardial disease had nine times more days in hospital than vaccinated persons with myocardial disease. Vaccination against pneumococcal infection had an additional effect with influenza vaccination in reducing hospitalization for chronic lung diseases; influenza vaccination alone did not have this effect. In conclusion, the influenza and pneumococcal-vaccine recipients were older and had significantly more chronic lung and heart conditions than the unvaccinated cohort. PMID- 12558335 TI - The incidence of influenza-associated hospitalizations in children in Germany. AB - Since new vaccines and anti-viral drugs for influenza have become available, collation of actual and country-specific epidemiological data is essential. Since respiratory syncytial virus (RSV) is a well known paediatric airway pathogen and some epidemiological data exist already, a comparison between influenza and RSV seems warranted. From July 1996 to June 2001 the naso-pharyngeal aspirates (NPA) of children from birth to 16 years of age, admitted to one of the two paediatric hospitals in Kiel, Germany, were investigated by a nine-valent multiplex reverse transcriptase PCR assay. NPA were investigated in 60.8 % of 3,469 children admitted with an acute respiratory tract infection. Community-acquired or nosocomial infections (in parentheses) due to influenza A were diagnosed in 122 (10) children, due to influenza B in 14 (2) and due to RSV in 325 (24) cases. Patients with influenza A (median 752 days) and influenza B (median 966 days) were older than patients with RSV (median 168 days). The spectrum of disease presentation was broader in influenza than in RSV. In each winter, admissions with influenza were less common than those with RSV. Influenza B only occurred in 2 of the 5 years. The cumulative, population-based incidences per 100,000 children 0-16 (0-5, >5-16) years of age were 53 (123, 22) for influenza A, 16 (30, 9) for influenza B and 165 (453, 4) for RSV. Cardiac conditions and asthma were the major risk factors for admission to hospital with influenza A (RR 9.8, 4.1) and RSV (8.5, 2.1) infections. Underlying conditions were most common in influenza B. Low gestational age doubled the risk for admission to hospital with influenza A infection, but did not show a dose-effect relationship as in RSV. The burden of influenza-positive hospitalizations was about one third that of RSV. The incidence was similar to reports from the United States. Targeting children with underlying conditions, especially cardiac conditions and asthma in the German immunization programme is appropriate, as long as no policy for vaccination of the general paediatric population exists. PMID- 12558336 TI - Age-specific seroprevalence of poliomyelitis, diphtheria and tetanus antibodies in Spain. AB - In 1996, a seroepidemiological study was undertaken in Spain, with the main aim of estimating the population's immunity against poliomyelitis, tetanus and diphtheria. A population-based cross-sectional study was conducted, covering the population aged 2-39 years. The sample was stratified by age and rural-urban environment, and informed consent obtained to take blood specimens from subjects attending phlebotomy centres. The study included 3,932 persons and the prevalence of antibodies against all three types of poliovirus exceeded 94% across all age groups. From a high of 96% in subjects under the age of 15 years, immunity against diphtheria steadily declined to a low of 32.3% in subjects aged 30-39 years. Similarly, tetanus antitoxin concentrations indicating basic protection were present in 98.9% of the under-14 years age group; thereafter, immunity declined, until reaching 54.6% in the 30-39 years age group. PMID- 12558337 TI - Epidemiology of measles, mumps and rubella in Italy. AB - A serosurvey for measles, mumps and rubella was conducted in Italy; incidence based on statutory notifications over the last three decades was also calculated. In Italy the diseases followed an endemic-epidemic pattern, with an incidence peak every 2-4 years, and had a limited reduction of incidence attributable to childhood immunization. Lower notification rates were observed in the Southern regions. This is possibly related to greater under notification in the South and is confirmed by our seroprevalence data. Incidence of measles and rubella and proportion of cases among young adults increased significantly in the three decades considered, but not for mumps. Serological data confirmed that these infections are still very frequent in Italy, without significant geographic variation in the country. In the age groups 2-4 and 5-9 years the percentage of individuals still susceptible to each virus was higher than 30%. The proportion of susceptible subjects older than 15 years was similar for the three infections (6.1, 11.7 and 8.8% for measles, mumps and rubella, respectively). The low vaccine coverage for rubella and measles in Italy has so far only partially affected the occurrence of the diseases. No impact of mumps vaccination is visible. The average number of deaths, for each disease, has decreased during the three study periods. Today the priority in Italy is to halt the progressive increase of the mean age of acquisition of the three infections, to eliminate differences in coverage among regions and to conform to European standards. This will be achieved through a combination of increasing MMR vaccine coverage before 2 years of age, implementing vaccination campaigns for low seroprevalence age groups, and/or introducing a second dose of MMR, depending on the level of current MMR coverage. PMID- 12558338 TI - Mumps outbreak in vaccinated children in Gipuzkoa (Basque Country), Spain. AB - A mumps outbreak occurred in a group of vaccinated children aged 3-4 years in San Sebastian (Gipuzkoa, Basque Country, Spain) in 2000 during the same period as a revaccination campaign against measles-mumps-rubella (MMR) was performed. The clinical cases were confirmed by viral culture, detection of viral RNA and/or specific IgM. Eighty-eight percent of the children had been vaccinated with the Rubini strain and the remainder with the Jeryl-Lynn strain. The attack rate was 47.9% (35 cases in 73 school-attending children of this age). The outbreak was caused by an H genotype strain of mumps virus which was circulating at the same time as a D genotype strain that caused sporadic cases. By sequencing the small hydrophobic (SH) gene, the strains of the clinical cases were identified as wild type mumps virus with heterologous genotypes in comparison to the vaccine strains used in our area. PMID- 12558339 TI - Rubella antibody loss rates in Korean children. AB - We followed students in eight elementary schools for rubella antibody from 1993 to 1996 (602 pairs) and 1996-9 (588 pairs) in Gyeonggi Province, Korea. We tested rubella IgG and administered rubella vaccine to the children with the titres < 10 IU/ml. The loss rates of rubella IgG during the follow-up periods were 14.3 and 15.8%, respectively. Among vaccinated groups, the loss rate was 18.8%, which was significantly higher than 13.8% of the mixture of natural and vaccine-induced immunity groups. The group that had the lower preceding antibody titre had a higher loss rate of 24.8% compared to 7.2% for the group whose titre was 40 IU/ml or above. In a multivariate analysis, age and gender were not related to antibody loss rate. Under this higher rubella antibody loss rate, in order to prevent congenital rubella syndrome, the immunization for women at childbearing age appears necessary until rubella can be eliminated or controlled. PMID- 12558340 TI - Regional and temporal changes in AIDS in Europe before HAART. AB - In a prospective observational study 4,485 patients from 46 clinical centres in 17 European countries were followed between April 1994 and November 1996. Information on AIDS-defining events (ADEs) were collected together with basic demographic data, treatment history and laboratory results. The centres were divided into four geographical regions (north, central, south-west and south east) so that it was possible to identify any existing regional differences in ADEs. The regional differences that we observed included a higher risk of all forms of Mycobacterium tuberculosis infections (Tb) and wasting disease in the south-west and an increased risk of infections with the Mycobacterium avium complex (MAC) in the north. In Cox multivariable analyses, where north was used as the reference group, we observed hazard ratios of 6.87, 7.77, 2.29 and 0.16 (P < 0.05 in all cases) for pulmonary Tb, extrapulmonary Tb, wasting disease and MAC respectively in the south-west. Pneumocystis carinii pneumonia (PCP) was less commonly diagnosed in the central region (RH = 0.51, 95% CI 0 32-0.79, P = 0.003) and most common in the south-east (RH = 1.04, 95% CI 0.71-1.51, P = 0.85). Comparisons with a similar 'AIDS in Europe' study that concentrated on the early phase of the epidemic reveal that most of the regional differences that were observed in the 1980s still persist in the mid-1990s. PMID- 12558341 TI - Monitoring hepatitis C virus infection among injecting drug users in the European Union: a review of the literature. AB - Hepatitis C virus (HCV) among injecting drug users (IDUs) is one of the European Union's (EU) major public health problems. This review examines the current state of knowledge regarding HCV among IDUs in EU countries. Studies published between January 1990 and December 2000, were identified through a computerized search (MEDLINE and EMBASE). Ninety-eight studies have reported prevalence for HCV among groups of IDUs in all EU countries except Luxembourg. The prevalence of anti-HCV ranged from 30 to 98%. Incidence rates ranged from 6.2 to 39.3 per 100 person years. This review provides a comprehensive examination of HCV infection among IDUs in the countries of the EU, and quite clearly demonstrates that the quality and epidemiological relevance of the studies published varies widely. Thus, the reported data may not reflect accurately the current or recent past prevalence of HCV among IDUs in the EU. A strategic approach to the surveillance of HCV among IDUs in the EU, utilizing robust and consistent methods, is required urgently. PMID- 12558342 TI - The contribution of transfusion to HCV infection in England. AB - The English HCV lookback programme has identified some individuals with transfusion-transmitted HCV infection. The path from the collection of donations from HCV-infected donors to the identification of infected recipients was constructed. The probability of different outcomes at each branch was derived from data collected during this programme. This path of probabilities was then used to produce a complete estimate of the number of recipients infected by blood transfusions (dead and alive at the end of 1995) by re-entry of blood components that fell out of the lookback at various steps prior to recipient testing, and entry of components from HCV-infected donations that were never identified for lookback. Less than 14,000 recipients were estimated to have been infected with HCV during the decade prior to the start of donation testing. Over 60% of these were expected to have died by the end of 1995. Transfusion has infected a large group of individuals. However, this group constitutes a very small, and declining, proportion of all HCV infections in the population. PMID- 12558343 TI - The role of solar ultraviolet irradiation in zoster. AB - Ultraviolet radiation (UVR) suppresses many aspects of cell-mediated immunity but it is uncertain whether solar UV exposure alters resistance to human infectious diseases. Varicella-zoster virus (VZV) causes varicella (chickenpox) and can reactivate from latency to cause zoster (shingles). The monthly incidence of chickenpox and zoster in a defined Polish population over 2 years was recorded and ground level solar UV was measured daily. There was a significant seasonality of UVR. Evidence of seasonal variation was found for all zoster cases and for zoster in males, with the lowest number of cases in the winter. The number of zoster cases with lesions occurring on exposed body sites (the face) demonstrated highly significant seasonality with a peak in July/August. Seasonal models for UVR and zoster cases showed similar temporal patterns. By contrast, for varicella, the maximum number of cases was found in March and the minimum in August/September, probably explained by the respiratory spread of VZV. It is tempting to speculate that the increase in solar UVR in the summer could induce suppression of cellular immunity, thus contributing to the corresponding rise in the incidence of zoster. PMID- 12558344 TI - Varicella in non-immune persons: incidence, hospitalization and mortality rates. AB - This study was conducted to estimate the varicella morbidity and mortality rates per age group among the non-immune population in France. Morbidity and mortality data for the years 1990-9 were derived from nationwide databases and surveillance systems. An incidence/prevalence model was designed to quantify the non-immune population per age group. The incidence of varicella in the non-immune population peaks during childhood and again in the 25-35 years age group. For children aged 1-4 years, adults aged 25-34 years and those older than 65 years, the hospitalization rates are respectively 235, 1,438 and 8,154 per 100,000 cases, and the death rates are respectively 7, 104 and 5,345 per million cases. Case fatality or case hospitalization rates were not evenly distributed among adults and increased dramatically with age. PMID- 12558345 TI - Immunity to chickenpox among school adolescents in Lebanon and options for vaccination. AB - Varicella infections cause substantial morbidity and mortality in adolescents and adults. The primary infection, chickenpox, results in lifelong immunity to chickenpox. A seroprevalence study carried on adolescents 15-18 years of age attending schools in Lebanon showed 96.6% immunity to varicella. The positive predictive value for immunity to chickenpox based on history alone was 97.4%, whereas the negative predictive value was 4.5%. Coming from a bigger family was a statistically significant predictor of immunity to chickenpox. In a developing country like Lebanon the merits and limitations of implementing universal varicella vaccination is discussed in relation to seroprevalence and socioeconomic factors. PMID- 12558346 TI - Evaluation of Lepto Dri Dot as a rapid test for the diagnosis of leptospirosis. AB - Lepto Dri Dot is a new card agglutination test developed by the Dutch Royal Tropical Institute for the rapid diagnosis of leptospirosis. We evaluated the test in field conditions in The Andaman Islands. Patients suspected of leptospirosis who attended three primary health centres were included in the study. The test results were compared with blood culture or microscopic agglutination tests on paired serum samples; 74 of 124 patients were diagnosed as having leptospirosis based on these criteria. Lepto Dri Dot had a sensitivity of 67.6% (50/74) and a specificity of 66.0% (33/50) during week 1. During weeks 2-4 the values increased to 85.5% (47/55) and 80% (40/50) respectively. An IgM ELISA was also performed on the serum samples for comparison and this was marginally less sensitive, but more specific, during the first week of illness. The positivity rates for the Dri Dot test during days 2-3, 4-5 and 6-7 were 53.1% (17/32), 75.0% (18/24) and 83.3% (15/18), respectively. The corresponding values for ELISA were 28.1% (9/32), 54% (13/24) and 77.8% (14/18). Both Dri Dot and ELISA showed good agreement with the standard diagnostic criteria after the first week of illness (kappa= 0.65 and 0.74, respectively). The overall concordance of the two tests was 89.5 % (kappa = 0.79). The test does not require special storage or sophisticated equipment and can be performed by relatively low skilled personnel. PMID- 12558347 TI - Predicting tuberculosis among migrant groups. AB - In industrialized countries migrants remain a high-risk group for tuberculosis (TB). Multiple linear regression analysis was used to determine the ability of indicators of TB incidence in the country of birth to predict the incidence of TB among migrants in Australia during 1997. World Health Organization total case notifications, new smear-positive case notifications and the estimated incidence of TB by country of birth explained 55, 69 and 87% of the variance in TB incidence in Australia, respectively. Gross national income of the country of birth and unemployment level in Australia were also significant predictors of TB in migrant groups. Indicators of the incidence of TB in the country of birth are the most important group-level predictors of the rate of TB among migrants in Australia. PMID- 12558348 TI - Local knowledge and attitudes about onchocerciasis in Oji-River local government area of Enugu State, Nigeria. AB - A total of 556 individuals in Agbalenyi Community of Oji-River local government area of Enugu State, Nigeria were studied using questionnaires, focus group discussions and key informant interviews to determine the socio-cultural factors affecting the transmission of onchocerciasis. The result revealed a low level of knowledge about the cause, prevention and complications of onchocerciasis. Respondents have developed a cultural system around the disease due to long exposure. The majority are aware of the disease which they recognize once their body starts itching or musculoskeletal pain develops but only 64.4, 34.0, 1.4 and 3.6%, respectively attributed chronic itching, nodules, bad vision and leopard skin to blackfly bite. Other perceived causes mentioned include ageing, the type of food eaten, farm work and 'bad blood'. Only 0.8% knew diagnosis could be made through skin snips. Neither Oji-River nor any of the river systems were associated with any of the complications of onchocerciasis. Significant differences in most socio-demographic characteristics were associated with differences in the level of knowledge about the disease. PMID- 12558349 TI - Salmonella and campylobacter contamination of raw retail chickens from different producers: a six year survey. AB - Between 1995 and 2000, a prospective survey was undertaken to investigate the levels of contamination of raw retail chickens (n = 1,127) with salmonella and campylobacter. The levels of contamination over the 6-year period were 11 % (95 % CI +/- 6.5%) for salmonella, and 57% (95% CI +/- 95%) for campylobacter. S. Bredeney (20%) and S. Enteritidis (18%) were the dominant serovars. Although salmonella contamination was higher than in an earlier survey we conducted (7%), since 1998 it has declined to 6%. Many S. Enteritidis isolates (43%) were associated with one large integrated poultry organization that appears to have successfully managed the contamination, and the serovar has not been isolated since 1998. Contamination ranged from 0 to 44% between different producers. There was no significant difference between producers contributing large and small numbers of samples, although some small producers had much poorer contamination rates than others. S. Bareilly, S. Bredeney, S. Enteritidis and S. Virchow showed associations with particular producers. Campylobacter contamination remains high. Contamination ranged from 47 to 81% between different producers. This study did not show a temporal association between contamination of chickens and human campylobacter infections, indicating that many cases of human campylobacteriosis, particularly during seasonal peaks, do not originate from chickens. Control measures that have reduced salmonella contamination have been largely ineffective against campylobacter and new interventions are needed. Most raw chickens are contaminated with these pathogens, and communicating the importance of minimizing this risk to caterers and the public is vital in reducing human infections. PMID- 12558350 TI - New world origins for haemoparasites infecting United Kingdom grey squirrels (Sciurus carolinensis), as revealed by phylogenetic analysis of bartonella infecting squirrel populations in England and the United States. AB - Phylogenetic analyses of bartonella have suggested divergence between bartonellae that infect mammals native to the Old and New Worlds. We characterized bartonella isolated from Eastern grey squirrels (Sciurius carolinensis) in the United States and from grey and red squirrels (Sciurus vulgaris) in the United Kingdom by nucleotide sequence comparison (gltA and groEL). Isolates from grey squirrels in the United States and the United Kingdom were identical, and most similar to Bartonella vinsonii, a species associated with New World rodents. A single and novel bartonella genotype was obtained from all 12 red squirrel isolates. Although grey squirrels were first introduced into the United Kingdom over 125 years ago, they continue to be infected solely by the bartonella associated with grey squirrels native to the United States. These results illustrate that exotic species may be accompanied by the introduction and maintenance, over many generations, of their microparasites. PMID- 12558351 TI - Evidence for insect transmission of rabbit haemorrhagic disease virus. AB - The spread of rabbit haemorrhagic disease (RHD) virus from quarantine on Wardang Island to mainland Australia in 1995 suggested that insects could be potential vectors. Field observations and laboratory experiments were conducted to address aspects of this hypothesis. Firstly, the variation in insect populations on the island during the field trials was examined. There was approximately a 1,000-fold increase in the number of bushflies, Musca vetustissima, shortly before the spread of the virus. Secondly, M. vetustissima were tested in the laboratory as potential vectors of RHD virus, and it was demonstrated that disease could be transmitted between rabbits by flies. Finally, 13 of 16 insect samples, collected from Wardang Island and from several sites on the mainland following the spread of virus off the island, were positive for the presence of RHD virus by a specific polymerase chain reaction (PCR). Only one sample contained sufficient infectious virus to kill a susceptible rabbit. These data, combined with previously published information on fly biology, suggested that flies, particularly bushflies, may be involved in the transmission of RHD virus. Other possible routes of spread were not assessed in this study. PMID- 12558352 TI - Epidemiological consequences of a pathogen having both virulent and avirulent modes of transmission: the case of rabbit haemorrhagic disease virus. AB - A number of pathogens cause chronic infection in survivors of acute disease and this is believed to be a common means of persistence, including for highly virulent agents. We present a model in which transmission from chronically infected hosts causes chronic infection in naive individuals, without causing acute disease--indeed 'protecting' against it. Thus the pathogen obtains the benefit of virulence (high transmission rate), but mitigates against the cost (high host mortality). Recent findings suggest that rabbit haemorrhagic disease virus (RHDV), a highly contagious and virulent pathogen, may also utilize this alternative, 'avirulent', mode of transmission. The model may resolve the paradox of how RHDV can be highly prevalent in some populations, in the absence of mortality. Differences in host demography determine whether avirulent transmission prevents large-scale mortality (as in most UK populations) or not. Other pathogens may exhibit similar behaviour and the implications for emerging diseases in general are discussed. PMID- 12558353 TI - The occurrence of five major Newcastle disease virus genotypes (II, IV, V, VI and VIIb) in Bulgaria between 1959 and 1996. AB - Partial sequence and restriction enzyme cleavage site analyses of the fusion protein gene were used to genotype 47 Newcastle disease virus strains isolated between 1959 and 1996 in Bulgaria. Viruses belonged to five major genotypes that appeared to be associated with epizootics characterized by temporal and/or geographical restrictions. Genotype IV viruses (responsible for the European branch of the first panzootic) dominated the scene up to the early 1980s, interspersed with sporadic outbreaks caused by genotype II (US strains causing pneumoencephalitis) viruses. Genotype V viruses (transmitted by psittacines from South America) were first shown in 1973 and persisted until the late 1980s. Genotype VI (earliest members from the Middle-East 1968/70 outbreaks) was represented by scattered isolations between 1974 and 1996. A genotype VIIb (recent Middle East epizootic) virus was isolated as early as in 1984. Newcastle disease epizootics in Bulgaria were highlighted by multiple infection with more than one genotype at any one time. PMID- 12558354 TI - The incidence and severity of adverse events affecting patients after discharge from the hospital. AB - BACKGROUND: Studies of hospitalized patients identify safety as a significant problem, but few data are available regarding injuries occurring after discharge. Patients may be vulnerable during this transition period. OBJECTIVE: To describe the incidence, severity, preventability, and "ameliorability" of adverse events affecting patients after discharge from the hospital and to develop strategies for improving patient safety during this interval. DESIGN: Prospective cohort study. SETTING: A tertiary care academic hospital. PATIENTS: 400 consecutive patients discharged home from the general medical service. MEASUREMENTS: The three main outcomes were adverse events, defined as injuries occurring as a result of medical management; preventable adverse events, defined as adverse events judged to have been caused by an error; and ameliorable adverse events, defined as adverse events whose severity could have been decreased. Posthospital course was determined by performing a medical record review and a structured telephone interview approximately 3 weeks after each patient's discharge. Outcomes were determined by independent physician reviews. RESULTS: Seventy-six patients had adverse events after discharge (19% [95% CI, 15% to 23%]). Of these, 23 had preventable adverse events (6% [CI, 4% to 9%]) and 24 had ameliorable adverse events (6% [CI, 4% to 9%]). Three percent of injuries were serious laboratory abnormalities, 65% were symptoms, 30% were symptoms associated with a nonpermanent disability, and 3% were permanent disabilities. Adverse drug events were the most common type of adverse event (66% [CI, 55% to 76%]), followed by procedure-related injuries (17% [CI, 8% to 26%]). Of the 25 adverse events resulting in at least a nonpermanent disability, 12 were preventable (48% [CI, 28% to 68%]) and 6 were ameliorable (24% [CI, 7% to 41%]). CONCLUSION: Adverse events occurred frequently in the peridischarge period, and many could potentially have been prevented or ameliorated with simple strategies. PMID- 12558355 TI - Individual and combined effects of age, breast density, and hormone replacement therapy use on the accuracy of screening mammography. AB - BACKGROUND: The relationships among breast density, age, and use of hormone replacement therapy (HRT) in breast cancer detection have not been fully evaluated. OBJECTIVE: To determine how breast density, age, and use of HRT individually and in combination affect the accuracy of screening mammography. DESIGN: Prospective cohort study. SETTING: 7 population-based mammography registries in North Carolina; New Mexico; New Hampshire; Vermont; Colorado; Seattle, Washington; and San Francisco, California. PARTICIPANTS: 329 495 women 40 to 89 years of age who had 463 372 screening mammograms from 1996 to 1998; 2223 women received a diagnosis of breast cancer. MEASUREMENTS: Breast density, age, HRT use, rate of breast cancer occurrence, and sensitivity and specificity of screening mammography. RESULTS: Adjusted sensitivity ranged from 62.9% in women with extremely dense breasts to 87.0% in women with almost entirely fatty breasts; adjusted sensitivity increased with age from 68.6% in women 40 to 44 years of age to 83.3% in women 80 to 89 years of age. Adjusted specificity increased from 89.1% in women with extremely dense breasts to 96.9% in women with almost entirely fatty breasts. In women who did not use HRT, adjusted specificity increased from 91.4% in women 40 to 44 years of age to 94.4% in women 80 to 89 years of age. In women who used HRT, adjusted specificity was about 91.7% for all ages. CONCLUSIONS: Mammographic breast density and age are important predictors of the accuracy of screening mammography. Although HRT use is not an independent predictor of accuracy, it probably affects accuracy by increasing breast density. PMID- 12558356 TI - Screening and surveillance for Barrett esophagus in high-risk groups: a cost utility analysis. AB - BACKGROUND: Once-in-a-lifetime screening for Barrett esophagus has been proposed for patients with gastroesophageal reflux disease (GERD), but there is little evidence of its cost-effectiveness. OBJECTIVE: 1) To determine the cost effectiveness of screening high-risk groups for Barrett esophagus and providing surveillance to patients with Barrett esophagus and dysplasia or to all patients with Barrett esophagus and 2) to compare the results with the cost-effectiveness of no screening or surveillance. DESIGN: A decision analytic model was developed to examine no screening or surveillance and screening and surveillance for Barrett esophagus with dysplasia only or Barrett esophagus without dysplasia every 2 to 5 years. Low- or high-grade dysplasia received surveillance every 6 or 3 months, respectively. DATA SOURCES: Published literature and the Health Care Financing Administration. TARGET POPULATION: 50-year-old white men with symptoms of GERD. TIME HORIZON: 50 years of age until 80 years of age or death. PERSPECTIVE: Third-party payer. OUTCOME MEASURE: Incremental cost-effectiveness ratio. RESULTS OF BASE-CASE ANALYSIS: Screening with surveillance limited to patients with Barrett esophagus with dysplasia required $10 440 per quality adjusted life-year (QALY) saved compared to no screening or surveillance. The incremental cost-effectiveness ratio of surveillance every 5 years in patients with Barrett esophagus without dysplasia compared to surveillance of patients with Barrett esophagus with dysplasia was $596 000 per QALY saved. RESULTS OF SENSITIVITY ANALYSIS: The annual incidence of adenocarcinoma must exceed 1 case per 54 patient-years of follow-up (1.9%) for surveillance of Barrett esophagus without dysplasia every 5 years to yield an incremental cost-effectiveness ratio less than $50 000 per QALY saved. CONCLUSIONS: Screening 50-year-old men with symptoms of GERD to detect adenocarcinoma associated with Barrett esophagus is probably cost-effective. However, subsequent surveillance of patients with Barrett esophagus but no dysplasia, even at 5-year intervals, is an expensive practice. PMID- 12558357 TI - Treatment of chronic hepatitis C in a state correctional facility. AB - BACKGROUND: Approximately 1 in 4 of the nearly 2 million individuals in state and federal correctional facilities are infected with hepatitis C virus (HCV). Currently, there are few reports of treatment outcomes of this common infection in this setting. OBJECTIVE: To describe HCV therapy in the incarcerated setting. DESIGN: Retrospective, descriptive observational study. SETTING: Rhode Island Department of Corrections, Cranston, Rhode Island. PATIENTS: 93 inmates with chronic HCV infection. INTERVENTION: Interferon-alpha with ribavirin. MEASUREMENTS: HCV RNA levels 6 months after treatment. RESULTS: Response rates are similar to previously published rates achieved in the community; 63% (50 of 79) of patients achieved viral clearance after 6 months of therapy, and 46% (26 of 57) achieved sustained response 6 months after treatment. CONCLUSION: The incarcerated population (which is disproportionately affected by addiction and psychiatric illness) can be effectively treated for HCV infection with interferon and ribavirin. The correctional setting may provide an opportunity to safely treat patients with these two challenging comorbid conditions. PMID- 12558358 TI - Complementary and alternative medical therapies: implications for medical education. AB - Increased use of complementary and alternative medicine (CAM) has made it imperative that these topics be included in medical education from the preclinical years through residency and beyond. There has been progress in this direction in recent years, with a steady increase in the number of medical schools that include CAM therapies in their curricula. There remains, however, a lack of clear goals and concrete suggestions for implementing these changes. This article examines the questions that arise when medical educators consider how to incorporate CAM therapies as an integral part of the medical curriculum. It offers practical suggestions for finding time in an already packed curriculum, getting started, including faculty and students in the process, and sustaining the initiative with the necessary administrative and institutional support. PMID- 12558360 TI - Professional organizations' position statements on physician-assisted suicide: a case for studied neutrality. AB - Position statements opposing legalization of physician-assisted suicide by organizations such as the American College of Physicians-American Society of Internal Medicine rightly emphasize that palliative care should be the standard of care for the dying, and that the inadequacies that exist in its delivery should be remedied. But such position statements generally understate the limitations of palliative care to alleviate some end-of-life suffering, and they do not provide adequate guidance about how physicians should approach patients with intractable suffering who are prepared to die. In this manuscript, we briefly present data about severe suffering before death for terminally ill patients, including those enrolled in hospice programs. We also review some of what is known about requests and responses for physician-assisted suicide in Oregon and in the rest of the United States. Preliminary data from Oregon suggest that legally sanctioned access to physician-assisted suicide is used by a very small number of patients and seems to be associated with improved delivery of hospice and palliative care. Physicians of good will, deep religious convictions, and considerable palliative care experience exist on both sides of the debate about legalization of physician-assisted suicide. In an effort to respect this diversity, and to encourage our profession to continue to struggle with the genuine dilemmas faced by some patients toward the end of their lives and by their families, we argue in favor of medical organizations' taking a position of studied neutrality on this contentious issue. PMID- 12558359 TI - Hepatitis C in the HIV-Infected Person. AB - Because of shared routes of transmission, hepatitis C virus (HCV) infection is common in HIV-infected persons, who have been experiencing increasing HCV-related morbidity and mortality since the advent of effective antiretroviral therapy. Infection with HIV appears to adversely affect the outcome of hepatitis C, leading to increased viral persistence after acute infection, higher levels of viremia, and accelerated progression of HCV-related liver disease. In addition, hepatitis C may affect the course and management of HIV infection. The medical management of hepatitis C in HIV-infected persons is complicated by immune suppression, potential drug interactions and toxicities, and other forms of liver disease. In addition, there is little published experience with the safety and efficacy of the best available anti-HCV medications in HIV-infected persons. Thus, current efforts must be directed at preventing HCV and HIV infections and applying the principles learned in treating persons with either infection to manage those with both. Future efforts should include studies of the pathogenesis of HCV infection in HIV-infected persons and large, prospective studies that demonstrate the optimal management of persons co-infected with HIV and HCV. Such efforts will help to eliminate HCV-related liver disease as an emerging threat to HIV-infected persons. PMID- 12558361 TI - Screening for type 2 diabetes mellitus in adults: recommendations and rationale. AB - This statement summarizes the current U.S. Preventive Services Task Force (USPSTF) recommendations on screening for type 2 diabetes in adults and updates the 1996 recommendations on this topic. The complete USPSTF recommendation and rationale statement on this topic, which includes a brief review of the supporting evidence, is available through the USPSTF Web site ( http://www.preventiveservices.ahrq.gov ) and the National Guideline Clearinghouse ( http://www.guideline.gov ) and in print through the Agency for Healthcare Research and Quality (AHRQ) Publications Clearinghouse (call 800-358-9295 or e mail mailto:ahrqpubs@ahrq.gov ). The complete information on which this statement is based, including evidence tables and references, is available in the accompanying article in this issue and in the summary of the evidence and systematic evidence review on this topic on the Web sites already mentioned. The summary of the evidence is also available in print through the AHRQ Publications Clearinghouse. PMID- 12558362 TI - Screening adults for type 2 diabetes: a review of the evidence for the U.S. Preventive Services Task Force. AB - BACKGROUND: Type 2 diabetes mellitus is associated with a heavy burden of suffering. Screening for diabetes is controversial. PURPOSE: To examine the evidence that screening and earlier treatment are effective in reducing morbidity and mortality associated with diabetes. DATA SOURCES: MEDLINE, the Cochrane Library, reviews, and experts, all of which addressed key questions about screening. STUDY SELECTION: Studies that provided information about the existence and length of an asymptomatic phase of diabetes; studies that addressed the accuracy and reliability of screening tests; and randomized, controlled trials with health outcomes for various treatment strategies were selected. DATA EXTRACTION: Two reviewers abstracted relevant information using standardized abstraction forms and graded articles according to U.S. Preventive Services Task Force criteria. DATA SYNTHESIS: No randomized, controlled trial of screening for diabetes has been performed. Type 2 diabetes mellitus includes an asymptomatic preclinical phase; the length of this phase is unknown. Screening tests can detect diabetes in its preclinical phase. Over the 10 to 15 years after clinical diagnosis, tight glycemic control probably reduces the risk for blindness and end stage renal disease, and aggressive control of hypertension, lipid therapy, and aspirin use reduce cardiovascular events. The magnitude of the benefit is larger for cardiovascular risk reduction than for tight glycemic control. The additional benefit of starting these treatments in the preclinical phase, after detection by screening, is uncertain but is probably also greater for cardiovascular risk reduction. CONCLUSIONS: The interventions that are most clearly beneficial during the preclinical phase are those that affect the risk for cardiovascular disease. The magnitude of additional benefit of initiating tight glycemic control during the preclinical phase is uncertain but probably small. PMID- 12558363 TI - The future of primary care. PMID- 12558364 TI - Payment and the future of primary care. PMID- 12558365 TI - Adopting more systematic approaches to hepatitis C treatment in correctional facilities. PMID- 12558366 TI - Emergency contraception. PMID- 12558367 TI - Sex differences in revascularization. PMID- 12558370 TI - Urinary catheters: a one-point restraint?. PMID- 12558372 TI - Reports of drug-induced thrombocytopenia. PMID- 12558373 TI - Primary care at a crossroads. AB - In response to concerns about the future of primary care, the Robert Wood Johnson Foundation sponsored a meeting in October 2001 in Glen Cove, New York, of 45 leaders in primary care and other health sectors. The purpose was to discuss the current and future challenges to primary care and to develop new and innovative ideas about how primary care might meet the needs of our current and future population. The premise of the meeting was that primary care is at a crossroads, which could lead to a new renaissance or a continued decline. Principles and ideas need to be identified to construct new primary care systems. As background for the meeting, 15 papers were written by experts in health care policy and primary care. Included in this supplement are revised versions of 4 of these papers, which present an overview of the current crisis, the implications of patients' views of primary care, the Chronic Illness Model as one possible solution, and a prediction of a tiering in the way primary care is delivered in the future. The final paper presents ideas and suggestions, based in part on the discussions at the meeting, about opportunities to move toward another renaissance of primary care. PMID- 12558374 TI - Primary care medicine in crisis: toward reconstruction and renewal. AB - Primary care is in crisis. Despite its proud history and theoretical advantages, the field has failed to hold its own among medical specialties. While the rest of medicine promises technology and sophistication, the basic model of primary care has changed little over the past half-century. Why has the transition from general practice to today's primary care been so difficult? Many of the causes of this struggle may lie within primary care itself, ranging from failure to articulate to the public (and insurers and policymakers) what value it, and it alone, can offer, to taking on an ever-broadening set of roles and responsibilities while all too often falling short of its promises. Perhaps most important, in the emerging health care system, the lack of a discrete definition of primary care has allowed managed care organizations and payers, among others, to define the role of primary care to suit their own interests. In response to a changing marketplace, political uncertainty, and shifting consumer expectations, primary care will need to reconstruct itself. The reconstruction will not be easy. Nevertheless, a process should begin that moves the field in the right direction. Building on its unique abilities, primary care can emerge as a redefined product that is attractive to patients, payers, and primary care practitioners alike. PMID- 12558375 TI - Defining the future of primary care: what can we learn from patients?. AB - From the earliest definitions of the term primary care to the most recent, all have stressed that primary care is predicated on a sustained relationship between patients and the clinicians who care for them. Primary care differentiates itself from other areas of medicine by attending to the whole person, in the context of the patient's personal and medical history and life circumstances, rather than focusing on a particular disease, organ, or system. Finally, the primary care physician plays a distinctive role in integrating the care that patients receive from within and outside of the primary care setting. Data obtained from patients over the past 15 years demonstrate that most Americans have a physician whom they consider to be their primary physician. This was the case well before the rules of managed care plans required patients to align themselves with a particular primary care physician and to allow that physician to coordinate all of their medical care. However, information from patients indicates that despite primary care relationships that endure over several years, the ideals of whole-person, integrated care are largely unmet in patients' primary care experiences. Moreover, considerable evidence indicates that the quality of primary care relationships has eroded over the past several years. This article highlights the relative strengths and weaknesses of primary care, as experienced and reported by patients, and posits three areas that must be addressed for primary care to live up to the ideals of sustained partnerships providing whole-person, integrated care. These three areas involve the use of teams in medicine, the establishment of meaningful primary care partnerships, and integration of care in a delivery system that patients experience as increasingly fragmented. PMID- 12558376 TI - Chronic illness management: what is the role of primary care?. AB - An estimated 99 million Americans live with a chronic illness. Meeting the needs of this population is one of the major challenges facing the U.S. health care system today and in the future. Dozens of studies, surveys, and audits have revealed that sizable proportions of chronically ill patients have not received effective therapy and do not have optimal disease control. The consistent findings of generally substandard care for many chronic conditions have spurred proposals that care be shifted to specialists or disease management programs. Published evidence to date does not indicate any clear superiority of these alternatives to primary care. The defining features of primary care (that is, continuity, coordination, and comprehensiveness) are well suited to care of chronic illness. A rapidly growing body of health services research points to the design of the care system, not the specialty of the physician, as the primary determinant of chronic care quality. The future of primary care in the United States may depend on its ability to successfully redesign care systems that can meet the needs of a growing population of chronically ill patients. PMID- 12558377 TI - Primary care in a new era: disillusion and dissolution?. AB - The current dilemmas in primary care stem from 1) the unintended consequences of forces thought to promote primary care and 2) the "disruptive technologies of care" that attack the very function and concept of primary care itself. This paper suggests that these forces, in combination with "tiering" in the health insurance market, could lead to the dissolution of primary care as a single concept, to be replaced by alignment of clinicians by economic niche. Evidence already exists in the marketplace for both tiering of health insurance benefits and corresponding practice changes within primary care. In the future, primary care for the top tier will cater to the affluent as "full-service brokers" and will be delivered by a wide variety of clinicians. The middle tier will continue to grapple with tensions created by patient demand and bureaucratic systems but will remain most closely aligned to primary care as a concept. The lower tier will become increasingly concerned with community health and social justice. Each primary care specialty will adapt in a unique way to a tiered world, with general internal medicine facing the most challenges. Given this forecast for the future, those concerned about primary care should focus less on workforce issues and more on macro health care financing and organization issues (such as Medicare reform); appropriate training models; and the development of a conception of primary care that emphasizes values and ethos, not just function. PMID- 12558378 TI - Primary care: the next renaissance. AB - Three decades ago, a renaissance helped create the foundations of primary care as we know it today. In recent years, however, new challenges have confronted primary care. We believe that the current challenges can be overcome and may, in fact, present an opportunity for a new renaissance of primary care to address the needs of our population. In this paper, we suggest seven core principles and a set of actions that will support a renaissance in, and a positive future for, primary care. The seven principles are 1) Health care must be organized to serve the needs of patients; 2) the goal of primary care systems should be the delivery of the highest-quality care as documented by measurable outcomes; 3) information and information systems are the backbone of the primary care process; 4) current health care systems must be reconstructed; 5) the health care financing system must support excellent primary care practice; 6) primary care education must be revitalized, with an emphasis on new delivery models and training in sites that deliver excellent primary care; and 7) the value of primary care practice must be continually improved, documented, and communicated. At the start of the 21st century, a vital, patient-centered primary care system has much to offer a rapidly changing population with increasingly diverse needs and expectations. If we keep the needs of persons and patients clearly in sight and design systems to meet those needs, primary care will thrive and our patients will be well served. PMID- 12558379 TI - Summaries for patients. Adverse events after hospital discharge. PMID- 12558380 TI - Summaries for patients. Screening adults for type 2 diabetes mellitus: recommendations from the U.S. Preventive Services Task Force. PMID- 12558381 TI - Summaries for patients. Treatment for hepatitis C virus infection among inmates. PMID- 12558382 TI - Summaries for patients. Cost-effectiveness of screening and surveillance for Barrett esophagus. PMID- 12558383 TI - Summaries for patients. The effects of age, breast density, and hormone therapy on the accuracy of screening mammograms. PMID- 12558384 TI - Screening for the risk for bleeding or thrombosis. AB - BACKGROUND: Numerous tests are available to assess patient risk for bleeding or thrombosis. Appropriate use of these tests must involve consideration of the clinical setting, disease prevalence, performance characteristics of the tests, cost, and consequences of false-positive and false-negative results. PURPOSE: To summarize information about coagulation testing in three common clinical settings: nonsurgical hospitalized patients, surgical patients, and patients having a first venous thromboembolic event. DATA SOURCES: All English-language studies identified in searches of MEDLINE (1966 to April 2002) and reference lists of key articles. STUDY SELECTION: All published studies of blood coagulation testing as routine diagnostic tests or in the preoperative care of patients reporting postoperative bleeding complications, and all published studies of patients with the factor V Leiden mutation reporting venous thromboembolic outcomes. DATA EXTRACTION: 5 observational studies of routine coagulation testing in nonsurgical hospitalized patients and 12 observational studies of preoperative coagulation testing, from which both sensitivity and specificity could be calculated. DATA SYNTHESIS: Test performance characteristics for the partial thromboplastin time in predicting postoperative hemorrhage were pooled by type of surgery. Likelihood ratios for positive and negative results were calculated for each group; 95% confidence intervals were calculated. Patients with prolonged partial thromboplastin times did not have a statistically significantly increased risk for postoperative complications. CONCLUSION: For nonsurgical and surgical patients without synthetic liver dysfunction or a history of oral anticoagulant use, routine testing has no benefit in assessment of bleeding risk. Routine testing after a first episode of venous thromboembolism is not recommended for most patients. PMID- 12558385 TI - Shape-persistent nanosize organometallic complexes: synthesis and application in a nanofiltration membrane reactor. AB - Shape-persistent multi(NCN-palladium and/or -platinum) complexes having one- (5 and 6), two- (1 and 2), and three-dimensional (3 and 4) geometries were prepared in moderate to good yields. Two different approaches were used to construct the multimetallic materials: (i) the construction of the multisite ligands followed by the permetalation step and (ii) selective and mild one-pot coupling of monometallic buiding blocks to a multifunctional shape-persistent organic core molecule. The first approach was used to prepare the palladated and/or platinated tris- (2) and bis(NCN-pincer) (5) complexes, while the second approach afforded the palladated and platinated octakis- (3) and dodecakis(NCN-pincer) (4) complexes. Complexes 1-6 were subjected to nanofiltration (NF) experiments in order to investigate the influence of rigidity and geometry on the retention of these molecules by NF membranes. For this purpose, the corresponding (NCN-Pt X)(n)() complexes (1c-4c, 5, and 6) were used since exposing these complexes to sulfur dioxide in solution resulted in the formation of bright orange complexes, allowing the use of UV/vis spectroscopy to accurately determine the concentrations of 1-6 in both retentate and permeate. Using the MPF-60 (MWCO = 400) NF-membrane, retention rates of 82.4 (6), 93.9 (1c), 98.7 (2c), 99.5 (3c), 99.6 (5), and >99.9% (4c) were found, while 2c and 4c in combination with the MPF 50 (MWCO = 700) NF-membrane were retained in 97.6 and 99.9%, respectively. A clear relationship is observed between the dimensions calculated by molecular modeling and the retention rates of 1-6. The one-dimensional bis(pincer-platinum) complex 5, however, shows an unexpectedly high retention rate (99.6%) that can be due to precipitation of the complex in the membrane (clogging of the membrane) and/or to the formation of larger aggregates near the membrane. In addition, comparison of 2 and 4 with flexible nickelated G0- and G1-dendrimers with similar dimensions proved that a high degree of rigidity in the backbone of macromolecular complexes indeed leads to more efficient retentions of these multimetallic materials by NF-membranes. PMID- 12558386 TI - Computational insight into the reaction intermediates in the glycosylation reaction assisted by donor heteroatoms. AB - We studied the geometric and electronic structure of several cations (9 and 18 23) that are usually proposed as intermediates for glycosylation reactions using DFT methods. For ions 9 and 18-23 the geometries obtained corresponded to an open ion (6b type) irrespective of the exocyclic heteroatom (S, Se, I) and the size of the cycle (furanoses as well as pyranoses). We studied how substituents bonded to the exocyclic heteroatom (R in SR and SeR) affect the structure of ions and found that the geometry approached that of episulfonium and episelenonium ions (6a type) when the substituents were electron donors. The tert-butyl group and the 2,4,6-trimethyl- and 2,4,6-trimethoxyphenyl and trimethylsilyl groups produced the strongest of these effects. The presence of an electron-withdrawing group (CO CF(3)) bonded to the hydroxymethyl group produced a similar but weaker effect. We analyzed this behavior in terms of molecular orbital interactions. PMID- 12558387 TI - Correlation between pK(a) and reactivity of quinuclidine-based catalysts in the Baylis-Hillman reaction: discovery of quinuclidine as optimum catalyst leading to substantial enhancement of scope. AB - The reactivity of a variety of quinuclidine-based catalysts in the Baylis-Hillman reaction has been examined, and a straightforward correlation between the basicity of the base and reactivity has been established, without exception. The following order of reactivity was established with pK(a)'s of the conjugate acids (measured in water) given in parentheses: quinuclidine (11.3), 3 hydroxyquinuclidine (9.9), DABCO (8.7), 3-acetoxyquinuclidine (9.3), 3 chloroquinuclidine (8.9), and quinuclidinone (7.2). The higher than expected reactivity of DABCO, based on its pK(a), was analyzed by comparing the relative basicity of DABCO and 3-acetoxyquinuclidine in DMSO. It was found that in aprotic solvent, DABCO was 0.6 pK(a) units more basic than 3-acetoxyquinuclidine, thus establishing a direct link between pK(a) of the amine and its reactivity. In contrast to previous literature work that reported the contrary, quinuclidine, which has the highest pK(a), was found to be the most active catalyst. The reaction profile with quinuclidine showed significant autocatalysis, which suggested that the presence of proton donors might further enhance rates. Thus, a series of additives bearing polar X-H bonds were investigated and it was found that methanol, triethanolamine, formamide, and water all provided additional acceleration. Methanol was found to be optimum, and the powerful combination of quinuclidine with methanol was tested with a host of aldehydes and Michael acceptors. Not only were the reactions more efficient and faster than previously reported, but now new substrates that were previously unreactive could be employed. Notable examples include the use of acetylenic aldehydes and the employment of vinyl sulfones, acrylamides, delta-lactones, and even alpha,beta unsaturated esters bearing a beta-substituent. PMID- 12558388 TI - Blue luminescent three-coordinate organoboron compounds with a 2,2' dipyridylamino functional group. AB - Two new three-coordinate organoboron compounds tris[p-(2,2'dipyridylamino) phenylduryl]borane (1) and tris[p-(2,2'dipyridylamino)biphenylduryl]borane (2) have been synthesized in good yields by using Pd-catalyzed Suzuki-Miyauya coupling reactions between tri(p-iododuryl)borane and the corresponding boronic acid. Both compounds display bright blue luminescence when irradiated by UV light. The emission band of both compounds is highly solvent-dependent, indicating the presence of a highly polarized excited state. These new boron compounds are stable under air in solution and the solid state, and are capable of binding to metal ions such as Zn(II) via the 2,2'-dipyridylamino chelating sites. The crystal structures of 1 and 2 have been determined. PMID- 12558389 TI - Effect of alkyl group size on the mechanism of acid hydrolyses of benzaldehyde acetals. AB - Hydrolyses of benzaldehyde acetals, PhCH(OR)(2), are specific hydrogen-ion catalyzed when R = methyl, n-butyl, but with secondary and tertiary alkyl derivatives, R = i-propyl, s-butyl, t-butyl, t-amyl, hydrolyses are general-acid catalyzed. The Bronsted alpha values for both secondary and tertiary alkyl groups are in the range: alpha = 0.57-0.61. A simple iterative procedure was developed to estimate the individual rate constants for general-acid catalysis by the diacid and monoacid forms of succinic acid buffer. Plots of log k(obs) (at [buffer] = 0 M) against pH are linear for the secondary and tertiary acetals, and plots of log k(H) for the H(3)O(+)-catalyzed reaction, (13)C and (1)H chemical shifts, and (1)J(CH) coupling constants against the Charton steric parameter, nu, for alkoxy groups are linear. The second-order rate constant, k(H), increases about 100-fold on going from R = Me to R = t-amyl, indicating the significant role of steric effects on reactivity. Steric effects upon (13)C NMR chemical shifts and coupling constants indicate that increasing the bulk of the alkoxy moiety increases the electron density at the carbon reaction center, which accelerates hydrolysis. Analysis of the Jencks-More-O'Ferrall free energy diagram for the reaction provides support for concerted proton transfer and C-O bond breaking in the transition state for hydrolyses of benzaldehyde acetals with secondary and tertiary alkyl groups in contrast to specific hydrogen catalysis with R = Me and n-Bu. All our results are consistent with rate-determining acid hydrolysis of benzaldehyde dialkyl acetals to hemiacetal intermediates that breakdown rapidly to benzaldehyde. PMID- 12558390 TI - Catalysis of the beta-elimination of HF from isomeric 2-fluoroethylpyridines and 1-methyl-2-fluoroethylpyridinium salts. Proton-activating factors and methyl activating factors as a mechanistic test to distinguish between concerted E2 and E1cb irreversible mechanisms. AB - Second-order rate constants, k(OH)(N), M(-)(1) s(-)(1), for the beta-elimination reactions of HF with 2-(2-fluoroethyl)pyridine (2), 3-(2-fluoroethyl)pyridine (3), and 4-(2-fluoroethyl)pyridine (4) in OH(-)/H(2)O, at 50 degrees C and mu = 1 M KCl, are = 0.646 x 10(-)(4) M(-)(1) s(-)(1), = 2.97 x 10(-)(6) M(-)(1) s(-)(1), and = 5.28 x 10(-)(4) M(-)(1) s(-)(1), respectively. When compared with the second-order rate constants for the same processes with the nitrogen-methylated substrates 1-methyl-2-(2-fluoroethyl)pyridinium iodide (5), 1-methyl-3-(2 fluoroethyl)pyridinium iodide (6), and 1-methyl-4-(2-fluoroethyl)pyridinium iodide (7), the methyl-activating factor (MethylAF) can be calculated from the ratio k(OH)(NCH)3/, and a value of 8.7 x 10(5) is obtained with substrates 5/2, a value of 1.6 x 10(3) with 6/3, and a value of 2.1 x 10(4) with 7/4. The high values of MethylAF are in agreement with an irreversible E1cb mechanism (A(N)D(E) + D(N)) for substrates 5 and 7 and with the high stability of the intermediate carbanion related to its enamine-type structure. In acetohydroxamate/acetohydroxamic acid buffers (pH 8.45-9.42) and acetate/acetic acid buffers (pH 4.13-5.13), the beta-elimination reactions of HF, with substrates 2 and 4, occur at NH(+), the substrates protonated at the nitrogen atom of the pyridine ring, even when the [NH(+)] is much lower than the [N], the unprotonated substrate, due to the high proton-activating factor (PAF) value observed: 3.6 x 10(5) for 2 and 6.5 x 10(4) for 4 with acetohydroxamate base. These high PAF values are indicative of an irreversible E1cb mechanism rather than a concerted E2 (A(N)D(E)D(N)) mechanism. Finally, the rate constant for carbanion formation from NH(+) with 2 is k(B)(NH)+ = 0.35 M(-)(1) s(-)(1), which is lower than when chlorine is the leaving group ( = 1.05 M(-)(1) s(-)(1); Alunni, S.; Busti, A. J. Chem. Soc., Perkin Trans. 2 2001, 778). This is direct experimental evidence that some lengthening of the carbon-leaving group bond can occur in the intermediate carbanion. This is a point of interest for interpreting a heavy-atom isotope effect. PMID- 12558391 TI - Efficient ring-opening reaction of epoxides and aziridines promoted by tributylphosphine in water. AB - Tributylphosphine was found to be an effective promoting reagent for the ring opening reaction of various epoxides and aziridines with nucleophile to produce corresponding anti-bifunctional products in moderate to excellent yields in water. PMID- 12558392 TI - Novel [4 + 2]-cycloaddition of 1-phenyl-1-benzothiophenium salts with dienes. Experimental evidence for a lack of aromaticity in the thiophene ring. AB - The [4 + 2]-cycloaddition reaction of 1-phenyl-1-benzothiophenium triflates has been conducted for the first time. [4 + 2]-Cycloaddition with dienes such as cyclopentadiene and 1,3-diphenylisobenzofuran occurs successfully to give cycloadducts. This result indicates that the C=C bond of the thiophene ring acts as a 2pi electron component in the cycloaddition reaction. Cycloadducts were formed in high yields with high stereoselectivity. However, the cycloaddition with other less reactive dienes such as 2,3-dimethyl-1,3-butadiene did not take place. The structure and stereochemistry of cycloadduct 2a were analyzed by NMR techniques. Furthermore, reaction of the cycloadducts with sodium methoxide in methanol gave the ring-opened products in high yields. PMID- 12558393 TI - First general approach to cyclohex-3-ene-1,1-bis(phosphonates) by Diels-Alder cycloaddition of tetraethyl vinylidenebis(phosphonate) to 1,3-dienes. AB - Tetraethyl vinylidenebis(phosphonate) (VBP) reacts smoothly with substituted 1,3 dienes at 90-110 degrees C without solvent to give the corresponding cyclohex-3 ene-1,1-bis(phosphonates) in good yields (60-85%). With nonsymmetrically substituted dienes, mixtures of regioisomers are obtained, the regioisomeric ratio being exclusively controlled by electronic effects. Danishefsky's diene allows tetraethyl 4-oxocyclohex-2-ene-1,1-bis(phosphonate) to be obtained in an 81% overall yield after the acid-catalyzed hydrolysis of the Diels-Alder cycloadduct. With 2,3-dimethoxy-1,3-butadiene, a mixture of regioisomeric dimethoxycyclohexene-1,1-bis(phosphonates) is formed by the VBP-catalyzed isomerization of the normal Diels-Alder cycloadduct. The mixture converges into tetraethyl 3,4-dimethoxycyclohex-2-ene-1,1-bis(phosphonate) at prolonged reaction times. PMID- 12558394 TI - A new selective cleavage of n,n-dicarbamoyl-protected amines using lithium bromide. AB - A mild and new procedure for the selective cleavage of an alkoxycarbonyl group (Boc, CBz) in N,N-dicarbamoyl-protected amino compounds is described. The method is based on the use of lithium bromide in acetonitrile and is compatible with a large range of other functionalities present in the substrates. Compared with other reported methodologies, the procedure is particularly useful for the Cbz selective cleavage in N,N-Ts,Cbz-diprotected amines. A rationalization of the selectivity supported by ab initio calculations is also presented. PMID- 12558395 TI - Desymmetrization of a centrosymmetric diepoxide: efficient synthesis of a key intermediate in a total synthesis of hemibrevetoxin B. AB - The preparation of an established intermediate in a total synthesis of hemibrevetoxin B is described. The acid-catalyzed cyclization of trans-4,5 epoxyoctane-2,7-dione exhibited a valuable mixture of kinetic and thermodynamic control: stereospecific epoxide opening was followed by equilibration of the products to provide the required trans-fused octahydropyrano[3,2-b]pyran ring system. Two-directional elaboration, by acetal substitution, ozonolysis, and sulfur ylide-mediated epoxidation, provided a centrosymmetric diepoxide. The key step of the synthesis was the first desymmetrization of a centrosymmetric molecule in natural product synthesis: Jacobsen asymmetric epoxide hydrolysis and acetonization provided the known synthetic intermediate in 97% yield and >95% ee over two steps. The exploitation of the center of symmetry of the AB ring system of the natural product contributed greatly to the efficiency (eight steps, 34% overall yield) of the synthesis. PMID- 12558396 TI - General scope of 1,4-diastereoselective additions to a 2(3H)-quinazolinone: practical preparation of HIV therapeutics. AB - The practical and highly diastereoselective syntheses of CF(3)-substituted dihydroquinazolinones via 1,4-additions of nucleophiles to chiral auxiliary substituted 2(3H)-quinazolinones is described. This methodology is applied to the syntheses of the NNRTIs (nonnucleoside reverse transcriptase inhibitors) DPC 961 (1) and DPC 083 (2), which are useful for the treatment of HIV (human immunodeficiency virus). The synthesis of DPC 961 (1) requires three steps, proceeds in >55% overall yield from the keto-aniline 9, and gives synthetic access to DPC 083 (2). In addition, the scope of the new diastereoselective 1,4 addition chemistry is investigated. The first preparation of DPC 961 (1) described in this paper is a derivatization fractional crystallization protocol. PMID- 12558397 TI - Rhodium-catalyzed addition of arylboronic acids to alkynyl aza-heteroaromatic compounds in water. AB - Alkynyl heteroaromatic compounds reacted with arylboronic acids to give addition products in the presence of a rhodium catalyst. The best results were obtained when a novel pyridine-substituted water-soluble phosphine ligand was used. The reactions proceed to give trisubstituted alkenes from various arylboronic acids and alkynyl heteroaromatic compounds with high regioselectivity. Only alkynes with a nitrogen atom in proximity to the triple bond were converted to the corresponding alkenes, as expected for a chelation-controlled addition. PMID- 12558398 TI - Preparation of 8-Amido-2-dimethylamino-1,2,3,4-tetrahydro-2-dibenzofurans and several fluorinated derivatives via [3,3]-sigmatropic rearrangement of O aryloximes. AB - Methodology to prepare 8-amido-2-amino-1,2,3,4-tetrahydro-2-dibenzofurans, analogues with a fluorine substituent incorporated in the 6-, 7-, and 9 positions, and a difluorinated analogue with fluorines in the 6- and 9-positions is described. The tetrahydrodibenzofuran ring systems are prepared by acid catalyzed [3,3]-sigmatropic rearrangement of O-aryloximes. Regioselective reactions to prepare the requisite O-aryloxime intermediates from commercially available fluorobenzene derivatives are discussed. PMID- 12558399 TI - Synthesis and properties of Bingel-type methanofullerene-pi-extended-TTF diads and triads. AB - Novel C(60)/pi-extended tetrathiafulvalene (exTTF) diads (12a-c) and triads [D(2)A (14a-c) and DA(2) (25, 27a-c)] have been synthesized by the Bingel cyclopropanation reaction of the respective exTTF-containing malonates and [60]fullerene. The reaction of exTTF-bismalonates with C(60) affords the respective C(60)-exTTF diads (26a-c) together with the triad C(60)-exTTF-C(60) (25, 27a-c) and a regioisomeric mixture of bisadducts (28b-c). Theoretical calculations (PM3) predict the favored geometry for triads 14a-c depending upon the orientation (up and down) of the 1,3-dithiole rings in the exTTFs, as well as the more stable regioisomers for the bisadducts 28. Cyclic voltammetry measurements reveal that C(60) and exTTF units do not intereact in the ground state. Compounds 26a-c and 27a-c are not electrochemically stable. A photoinduced electron transfer leading to the formation of the radical pair (C(60)(-) exTTF(*+)) has been observed for compounds 14a-c. PMID- 12558401 TI - A short olefin metathesis-based route to enantiomerically pure arylated dihydropyrans and alpha, beta-unsaturated delta-valero lactones. AB - The synthesis of arylated dihydropyrans and unsaturated lactones starting from enantiomerically pure alpha-hydroxy ketones (prepared by an enzyme-catalyzed benzoin condensation) is described. The key steps are a highly diastereoselective addition of vinyl metal compounds under chelate control and a ruthenium-catalyzed ring-closing olefin metathesis reaction. Elucidation of the relative configuration of the final products was achieved by NOE experiments. PMID- 12558400 TI - An asymmetric synthesis of aza analogues of the tricyclic skeleton of daphnane and the ABC ring system of phorbol. AB - An asymmetric synthesis of aza analogues of the ABC ring system of phorbol and related compounds containing the 5-7-6-fused framework of daphnane involved construction of the central seven-membered ring by a regioselective reduction of a chiral imide and cyclization with trifluoromethanesulfonic acid. Subsequent demethylation and oxidative dearomatization of ring C afforded an enantiopure dienone 20 with the same relative and absolute configuration at the 9- and 10 positions of the phorbol skeleton. PMID- 12558402 TI - A facile synthesis of enantiopure tricyclic furanyl and pyranyl derivatives via tungsten-mediated cycloalkenation and Diels-Alder reaction. AB - We report the synthesis of chiral furanyl and pyranyl dienes 1 and 2 based on cycloalkenation of chiral tungsten alkynol complexes. These two dienes bear a chiral 1,3-dioxolane group to control diastereoselective Diels-Alder reactions with electron-deficient olefins. The chiral 1,3-dioxolane substituents of the cycloadducts were degraded into hydrogen atoms to make these molecules possess common furan and pyran rings. Dienes 1 and 2 are good building blocks for enantiopure forms of tricyclic oxygen compounds. PMID- 12558403 TI - Novel pathways for oxygen insertion into unactivated C-H bonds by dioxiranes. Transition structures for stepwise routes via radical pairs and comparison with the concerted pathway. AB - The oxygen insertion into C-H bonds (of methane, isobutane, and acetone) by dioxiranes (parent dioxirane and dimethyldioxirane) to give alcohols was studied with the DFT theory, using both restricted and unrestricted B3LYP methods, and 6 31G(d) and 6-311+G(d,p) basis sets to evaluate the feasibility of stepwise mechanisms and their competition with the concerted counterpart. Confirming previous results by other authors, we have located, with the RB3LYP method, concerted TSs in which the oxygen bound to be inserted interacts very strongly with the hydrogen atom and very weakly with the carbon atom of the C-H bond. These TSs nicely explain all the experimental observations (e.g., configuration retention at the chiral centers), but all of them exhibit an RHF --> UHF wave function instability that preclude considering them as genuine transition structures. We also were able to characterize, with UB3LYP methods, two alternative two-step processes that can lead to final products (alcohol + carbonyl compound) via singlet radical pair intermediates. For the first step of both processes we located genuine diradicaloid TSs, namely, TSs rad,coll and TSs rad,perp, that have stable wave functions. In TSs rad,coll the alkane C-H bond tends to be collinear with the breaking O(1)- - -O(2) bond while in TSs rad,perp the alkane C-H bond is almost perpendicular to the O(1)- - -O(2) bond. The first step, of both processes, can represent an example of a "molecule induced homolysis" reaction: collision between alkane and dioxirane brings about the homolytic cleavage of the dioxirane O-O bond and the hydrogen abstraction follows afterward to produce the diradicaloid TS that then falls down to a singlet radical pair. This hypothesis was fully confirmed by IRC analysis in the case of TSs rad,coll. The possible pathways that lead from the intermediate radical pair to final products are discussed as well as the hypothesis that the radical collinear TSs may collapse directly to products in a "one-step nonconcerted" process. However, diradical mechanisms cannot explain the experimental data as satisfactorily as the concerted pathway does. As for computational predictions about competition of diradical vs concerted mechanisms, they strongly depend (i) on the alkane C-H type, (ii) on whether gas phase or solution is considered, and (iii) on the basis set used for calculations. In short, the concerted TS benefits, with respect to the corresponding diradicaloid TSs, of alkyl substitution at the C-H center, solvation effects, and basis set extension. Actually, in the case of DMD reactions with methane and acetone, the diradicaloid TSs are always (both in gas phase and in solution and with both the basis sets used) strongly favored over their concerted counterpart. In the case of DMD reaction with isobutane tertiary C-H bond the large favor for the diradicaloid TSs over the concerted TS, predicted in gas phase by the B3LYP/6-31G(d) method, progressively decreases as a result of basis set extension and introduction of solvent effects: the higher theory level [B3LYP/6-311+G(d,p)] suggests that in acetone solution TS conc has almost the same energy as TS rad,perp while TS rad,coll resides only 2 kcal/mol higher. PMID- 12558404 TI - Acetal-vinyl sulfide cyclization on sugar substrates: effect of structure and substituent. AB - A range of 2-deoxyfuranoside and -pyranoside derivatives were fashioned into derivatives that carry a vinyl or propenyl side chain. Extension of the alkene by a Suzuki cross-coupling reaction with 1-bromo-1-(phenylthio)ethene gave thioenol ethers as the cyclization substrates. The treatment of these substrates with BF(3).Et(2)O in tert-butylmethyl ether below 0 degrees C induced cyclization to optically active bicyclic ethers. If the cyclizations are carried out in toluene as the solvent, the isomerization of the terminal thioenol ether to the inner thioenol ether can take place prior to the cyclization. The cyclization reactions can be impeded by steric and electronic factors. The opening of the bicyclic ethers could be illustrated with the base-induced conversion of the ketone 53 to the cyclooctenone 54. PMID- 12558405 TI - Synthesis of novel cross-conjugated dendritic fluorophores containing both phenylenevinylene and phenyleneethynylene moieties. AB - New dendrons and dendrimers with dendritic arms composed of alternate phenyleneethynylene and phenylenevinylene moieties have been efficiently synthesized using orthogonal and convergent syntheses that combine Sonogashira and Horner-Wadsworth-Emmons reactions. Two different iterative routes have been developed that allow specific control over the placement of double and triple bonds within the interior of the dendrimers. A preliminary study of the UV and photoluminescence (PL) properties of the resulting compounds is also described. All of the examples prepared are blue-luminescent. PMID- 12558406 TI - Diarylamino groups as photostable auxofluors in 2-benzoxazolylfluorene, 2,5 diphenyloxazoles, 1,3,5-hexatrienes, 1,4-distyrylbenzenes, and 2,7 distyrylfluorenes. AB - The relationship of structure to optical spectral properties was determined for five types of fluors in a search for an optimum-wavelength shifter to be used as part of the detection systems for high-energy particles from accelerators. In a search for photostable fluors to serve as waveshifters in plastic fibers it was found that the wavelengths of interest, absorption max 410 +/- 10 nm and fluorescence emission max 480 +/- 20 nm, along with other properties, such as high solubility and short fluorescence decay time, could be obtained from fluorophors composed of aromatic rings and vinyl groups only by using amino groups as auxochromes to give bathochromic shifts of wavelengths. Since primary, monoalkyl, and dialkylamino groups were not sufficiently photostable, a number of fluorophores bearing diarylamino groups were investigated. Syntheses of the fluors made use of the Buchwald amination, an improved version of the Emmons Horner reaction, and other common reactions. The fluor types were the following: a 2-benzoxazolyl-7-(4-diarylamino)fluorene 7, 2-(4-cyanophenyl)-5-(4 aminophenyl)oxazoles 14 and 20, 1,3,5-hexatrienes 24a-d and 26a-c, 1,4 distyrylbenzenes 31d-g and 32a-e, and 2,7-distyrylfluorenes 40a,d-e. The unsymmetrical fluors 7, 14, and 20 were not as bright as the best hexatrienes, distyrylbenzenes, and distyrylfluorenes, which were all symmetrical. Where the 1,6-diaryl-1,3,5-hexatrienes 24a-d had high fluorescence quantum yield (Phi(f)), the 1,1,6,6-tetraryl-1,3,5-hexatrienes 26a-c had both lower epsilon and Phi(f). Where the 1,4-distyrylbenzenes 31d-g had high Phi(f), the 1,4-bis(2 phenylstyryl)benzenes 32a-e had Phi(f) = 0. Diarylamino groups as auxofluors conferred higher photochemical stability than dialkylamino groups on similar fluorophores. The 1,4-distyrylbenzenes 31d,e and the 2,7-distyrylfluorenes 40d,ehad the most desirable properties overall, which included fast decay times of 2 ns. Computer simulations predicted absorption and emission wavelengths fairly well, but were of little help for the prediction of brightness, stability, Phi(f), or decay time. PMID- 12558407 TI - Naphthalene analogues of lignans. AB - The methodology for the synthesis of podophyllotoxin and thuriferic acid-type lignans has been applied to derivatives carrying a naphthalene moiety. Starting from the 1,3-dithiane of 2-naphthaldehyde afforded the expected analogues in the 2,1-naphthalene series. The preferred conformations of these compounds are influenced by the bulky naphthalene system. By contrast, 1,8-bridged products were obtained from the 1,3-dithiane of 1-naphthaldehyde. In this series, polycyclic naphthalene lignan analogues were isolated after deprotection and/or desulfurization reactions. The cyclizations produced in this process are due to the proximity between the 3,4,5-trimethoxyphenyl moiety and the reacting C-2 of the 1,3-dithiane ring. PMID- 12558408 TI - Catalytic specificity exhibited by p-sulfonatocalix[n]arenes in the methanolysis of N-acetyl-l-amino acids. AB - Specific acid catalysis of p-sulfonatocalix[n]arenes (n = 4, Calix-S4; n = 6, Calix-S6; n = 8, Calix-S8) was observed in the alcoholysis of N-acetyl-l-amino acids in methanol. The methanolysis rates of basic amino acid substrates (His, Lys, and Arg) were markedly enhanced in the presence of Calix-Sn, as compared with rates observed with p-hydroxybenzenesulfonic acid (pHBS), which is a noncyclic analogue of Calix-Sn. This catalytic effect of Calix-Sn was not observed for the methanolysis of Phe, Tyr, and Trp substrates. On the other hand, (1)H NMR experiments following the effect of Calix-Sn on N-acetyl-l-amino acid substrates in CD(3)OD showed that the spectrum of a mixture of the His substrate with Calix-Sn was significantly different from the combined spectra of the respective compounds. These changes in spectra support the formation of an inclusion complex of Calix-Sn with basic amino acids. Furthermore, it was obvious that methanolysis of the His substrate catalyzed by Calix-S4 and Calix-S6 obeyed Michaelis-Menten kinetics. These results indicate that the catalytic activity of Calix-Sn originates from its forming a complex with specific substrates (basic amino acids), similar to enzymatic reactions. PMID- 12558409 TI - Phosphoramidites are efficient, green organocatalysts for the Michael reaction. Mechanistic insights into the phosphorus-catalyzed Michael reaction of alkynones and implications for asymmetric catalysis. AB - Hexamethylphosphorous triamide (HMPT) and other phosphoramidites and phosphites have been found to be efficient catalysts for the Michael reaction of alkenones and alkynones with malonates, alpha-cyano esters, beta-keto esters, and nitro compounds. The relatively nontoxic, easily hydrolyzed HMPT catalyzes the Michael reaction within seconds at room temperature in the absence of a solvent, and the reaction is worked up simply by removing the catalyst in vacuo. The Michael reactions of alkynones, unlike those of alkenones, are shown to be irreversible. The implications for asymmetric catalysis are discussed. PMID- 12558410 TI - Substituent effects on the acidity of weak acids. 3. Phenols and benzyl alcohols. AB - The gas-phase acidities of meta- and para-substituted phenols have been calculated at the B3LYP/6-311+G, MP2/6-311+G, MP2/6-311++G, and MP2/6 311+G(2df,2pd) theoretical levels. The larger basis sets give the more satisfactory DeltaHacid values that are correlated with the observed acidities with a slope close to unity. They are systematically about 2 kcal/mol too small. The acidities of most substituted phenols are linearly related to those of the corresponding substituted benzoic acids and benzyl alcohols. Here, the substituent effect is a Coulombic interaction (field effect) of the distributed charge in the benzene ring with the negative charge of the anionic center. The exceptions are strong para-substituted pi-acceptors such as NO(2) and CHO, and to a smaller extent, CN and CF(3). Here there is direct charge transfer from the phenoxide oxygen to the substituent. PMID- 12558411 TI - A new suzuki-heck-type coupling cascade: indeno[1,2,3]-annelation of polycyclic aromatic hydrocarbons. AB - Under palladium catalysis, o-bromobenzeneboronic acid can be coupled with 1 bromonaphthalene (6) and with oligocyclic bromoarenes to furnish indeno-annelated polycyclic aromatic hydrocarbons 1-4 and 25 in a single operation in moderate to good yields (27-87%). Alternatively, o-dibromoarenes and 1,2-dibromocycloalkenes can be cross-coupled with 1-naphthaleneboronic acid under the same conditions to yield analogous products (6-87%), and indenocorannulene (19) can be prepared likewise in a single step from pinacol corannuleneboronate (18) (40%). PMID- 12558412 TI - Rapid and amenable suzuki coupling reaction in water using microwave and conventional heating. AB - It is possible to prepare biaryls in good yield very rapidy (5-10 min) on small (1 mmol) and larger (10-20 mmol) scales from aryl halides and phenylboronic acid using water as a solvent and palladium acetate as catalyst. The reaction can be performed equally well using microwave and conventional heating, showing that using these conditions probably no nonthermal microwave effects are associated with the impressive speed of the reaction. PMID- 12558413 TI - Investigation of the transition-metal- and acid-catalyzed reactions of beta-(N tosyl)amino diazo carbonyl compounds. AB - A series of beta-(N-tosyl)amino diazo carbonyl compounds have been prepared by nucleophilic condensation of N-tosylimines with acyldiazomethanes. The diazo decomposition of these diazo carbonyl compounds under various catalytic conditions, including Rh(II) carboxylates, Cu(I) and Cu(II) complexes, PhCO(2)Ag/Et(3)N, TsOH, and SnCl(2).2H(2)O, has been investigated. It was found that, in most cases, the diazo decomposition gave preferentially 1,2-aryl migration product, but 1,2-hydride migration predominated when PhCO(2)Ag/Et(3)N was the catalytic system. Hammett correlation has been applied in the analysis of the electronic effects of 1,2-aryl migration. The factors that govern the migratory preference and the mechanistic aspects of the reaction are discussed. PMID- 12558414 TI - Iron-mediated preparation of vinylcyclopropanecarboxylates: scope, mechanism, and applications. AB - The addition of stabilized carbon nucleophiles to tricarbonyl(1 methoxycarbonylpentadienyl)iron(1+) cation (1a) proceeds via attack at C2 on the face of the ligand opposite the Fe(CO)(3) group to generate tricarbonyl(pentenediyl)iron complexes 2. Oxidation of complexes 2 affords vinylcyclopropanecarboxylates in good yield. In general, the relative stereochemistry about the cyclopropane ring reflects reductive elimination with retention of configuration. In cases where the C2 substituent is bulky (i.e., 2b) the major cyclopropane product 9b represents ring closure with inversion at C3. A mechanism involving pi-sigma-pi rearrangement of the initially oxidized (pentenediyl)iron species is proposed to account for these results. Experiments which probe the stereochemistry of deuterium labeling in the vinyl group of the vinylcyclopropanecarboxylate products were carried out, and these results are consistent with the proposed mechanism. This methodology for the preparation of vinylcyclopropanecarboxylates was applied to the synthesis of 2-(2' carboxycyclopropyl)glycines (+)-22 and (-)-23 and the cyclopropane triester (-) 26. PMID- 12558415 TI - Transient azomethine-ylides from a stable amino-carbene and an aldiminium salt. AB - Catalytic amounts of a protic reagent such as tert-butyl alcohol promote the isomerization of a stable amino-aryl-carbene into a transient azomethine ylide. Deprotonation of an alkyl-aldiminium salt also leads to a transient azomethine ylide, but labeling experiments rule out the transient formation of the corresponding amino-alkyl-carbene. The potential hypersurface between model amino carbene, aziridine, and azomethine ylide is investigated. PMID- 12558416 TI - Chiral Lewis acid-catalyzed asymmetric Baylis-Hillman reactions. AB - An effective chiral Lewis acid-catalyzed asymmetric Baylis-Hillman reaction is described. Good to high enantioselectivities were obtained using 3 mol % chiral catalyst. Novel camphor-derived dimerized ligands were prepared from the condensation of (+)-ketopinic acid with the corresponding diamines and hydrazine under acidic conditions. When alpha-naphthyl acrylate was used as a Michael acceptor, the reaction is complete within 20 min with high stereoselectivity and in reasonable chemical yields. PMID- 12558417 TI - Synthesis of 3,4-disubstituted isoquinolines via palladium-catalyzed cross coupling of 2-(1-alkynyl)benzaldimines and organic halides. AB - The palladium-catalyzed cross-coupling of readily available N-tert-butyl-2-(1 alkynyl)benzaldimines and aryl, allylic, benzylic, alkynyl halides, as well as a vinylic halide, provides a valuable new route to 3,4-disubstituted isoquinolines with aryl, allylic, benzylic, 1-alkynyl, and vinylic substituents, respectively, in the 4-position. The reaction appears to require an aryl group on the end of the acetylene furthest from the imine functionality. The reaction conditions have been optimized, and reasonably good yields have been obtained. PMID- 12558418 TI - A short diastereoselective synthesis of the putative alkaloid jamtine, using a tandem pummerer/mannich cyclization sequence. AB - Treatment of 2-phenylhex-5-enal with benzylamine followed by sequential reaction with ethylthioacetyl chloride and sodium periodate oxidation afforded a E/Z mixture of alpha-sulfinylamides. As anticipated from a 4pi-conrotatory mechanism, cyclization of each olefin afforded fused isoquinoline lactams as single diastereomers epimeric at the ethylthio position without any cross contamination. Some preliminary studies were directed toward the synthesis of mesembrine using a 3,4-dimethoxy aryl group. In this case, the Z-enamide prefers to undergo electrophilic aromatic substitution to give a substituted azepinone as the preferred product in 87% yield. In contrast, the E-enamide isomer provided the desired hydroindolone. The convergency and stereochemical control associated with the tandem Pummerer /Mannich cyclization make it particularly suited for the assembly of jamtine, a tetrahydroisoquinoline alkaloid reputed for its therapeutic properties. The key step in the synthesis involves a domino thionium/N-acyliminium ion cyclization to provide the tricyclic ring skeleton 27a as the major diastereomer. Deprotonation of 27a with NaH gave 28a, which contains the fully assembled skeleton of jamtine. Completion of the synthesis entailed installation of the double bond and reduction of the lactam. Oxidation of a synthetic sample of jamtine with MCPBA afforded the corresponding N-oxide, which does not match the spectral data reported in the literature for this alkaloid. Our synthetic efforts raise the possibility of a revision of the earlier assignment. PMID- 12558419 TI - Memory of chirality generated by spontaneous crystallization and asymmetric synthesis using the frozen chirality. AB - Asymmetric synthesis using frozen chirality generated by spontaneous crystallization was performed. Achiral asymmetrically substituted imide with a tetrahydronaphthyl group on the nitrogen atom crystallized in a chiral fashion, with space group P2(1)2(1)2(1). The molecular chirality generated by spontaneous crystallization was retained in cold THF. The half-life determined on the basis of decreasing optical activity followed by CD spectrometer was 7.8, 33.1, and 150.0 min at -20, -30, -40 degrees C, respectively. The energy barrier (DeltaG()) of racemization was calculated with the temperature dependence of the kinetic constant to be 18.24-18.36 kcal mol(-)(1) at 233-253 K. The memorized frozen chirality was transferred to permanent optically active alcohols by nucleophilic addition with n-buthyllithium. PMID- 12558420 TI - Stable enols of amides ArNHC(OH)=C(CN)CO(2)R. E/Z enols, equilibria with the amides, solvent effects, and hydrogen bonding. AB - The structures of anilido cyano(fluoroalkoxycarbonyl)methanes ArNHCOCH(CN)CO(2)R, where R = CH(2)CF(3) or CH(CF(3))(2), Ar = p-XC(6)H(4), and X = MeO, Me, H, or Br, were investigated. In the solid state, all exist as the enols ArNHC(OH)=C(CN)CO(2)R 7 (R = CH(2)CF(3)) and 9 (R = CH(CF(3))(2)) with cis arrangement of the hydrogen-bonded ROC=O.HO moiety and a long C1=C2 bond. The product composition in solution is solvent dependent. In CDCl(3) solution, only a single enol is observed, whereas in THF-d(8) and CD(3)CN, two enols (E and Z) are the major products, and the amide is the minor product or not observed at all (K(Enol) 1.04-9 (CD(3)CN, 298 K) and 3 to >/=100 (THF, 300 K)). The percentage of the amide and the Z-enol increase upon an increase in temperature. In all solvents, the percent enol is higher for 9 than for 7. In CD(3)CN, more enol is observed when the aryl group is more electron-donating. The spectra in DMSO-d(6) and DMF-d(7) indicate the presence of mostly a single species, whose spectra do not change on addition of a base and is ascribed to the anion of the ionized carbon acid. Comparison with systems where the CN is replaced by a CO(2)R group (R = CH(2)CF(3), CH(CF(3))(2)) shows a higher percentage of enol for the CN substituted system. Intramolecular (to CO(2)R) and intermolecular hydrogen bonds determine, to a significant extent, the stability of the enols, their Z/E ratios (e.g., Z/E (THF, 240 K) = 3.2-4.0 (7) and 0.9-1.3 (9)), and their delta(OH) in the (1)H spectra. The interconversion of Z- and E-enol by rotation around the C=C bond was studied by DNMR, and DeltaG() values of >/=15.3 and 14.1 +/- 0.4 kcal/mol for Z-7 and Z-9 were determined. Features of the NMR spectra of the enols and their anions are discussed. PMID- 12558421 TI - Comprehensive and uniform synthesis of all naturally occurring phosphorylated phosphatidylinositols. AB - Studies of cellular signal transduction mechanisms involving receptor-mediated generation of inositol phosphates and phosphorylated phosphatidylinositols require easy access to these naturally occurring products. Although numerous synthetic methods have been developed during the past decade, most of these methods suffer from excessive length and lack of generality. In this work we describe the comprehensive and uniform synthesis of all naturally occurring phosphatidylinositols such as phosphatidylinositol, phosphatidylinositol 3 phosphate, 4-phosphate, 5-phosphate, 3,4-bisphosphate, 3,5-bisphosphate, 4,5 bisphosphate, and 3,4,5-trisphosphate, featuring both saturated and unsaturated fatty acid chains. PMID- 12558422 TI - Coupling reactions of alpha-(N-carbamoyl)alkylcuprates with enol triflates derived from cyclic beta-keto esters: A facile approach to gamma-carbamoyl alpha,beta-enoates. AB - alpha-(N-Carbamoylalkyl)cuprates couple with enol triflates derived from carbocyclic and heterocyclic (i.e., piperidinones) beta-keto esters. Product yields are higher with the alkyl(cyano)cuprates [i.e., RCu(CN)Li, 56-93%] than with the dialkylcuprate reagents (i.e., R(2)CuLi.LiCN). An enol nonaflate works as well as the corresponding enol triflate. A facile synthetic route to gamma amino alpha,beta-enoates not readily prepared from gamma-keto-alpha,beta-enoates is thus established. The gamma-amino-alpha,beta-enoates, available via N-Boc deprotection, can be cyclized to annulated pyrrolin-2-ones. PMID- 12558423 TI - Highly enantioselective mukaiyama aldol reaction of alpha,alpha-dichloro ketene silyl acetal: an efficient synthesis of a key intermediate for diltiazem. AB - An efficient synthesis of methyl (2R,3S)-3-(4-methoxyphenyl)glycidate (-)-2, a key intermediate for diltiazem (1), has been developed on the basis of the highly enantioselective Mukaiyama aldol reaction of p-anisaldehyde (4a) with alpha,alpha dichloro ketene silyl acetal 5. Thus, the reaction using a stoichiometric amount of chiral oxazaborolidinone catalyst 12a proceeded to excellent yield (83%) and high enantioselectivity (96% ee), together with the chiral ligand 13a in nearly quantitative recovery. The reaction using a substoichiometric amount of 12e (20 mol %) also proceeded to excellent yield (88%), with somewhat lower enantioselectivity (77% ee). The aldol product 3a thus obtained was easily converted to (-)-2 in excellent yield (80%) and high optical purity (>99% ee). The highly enantioselective Mukaiyama aldol reaction with 5 catalyzed by 12a proved to be applicable to various aldehydes. An efficient preparation of 5 from inexpensive starting materials was also described. PMID- 12558424 TI - Synthesis of 4-(1-alkenyl)isoquinolines by palladium(II)-catalyzed cyclization/olefination. AB - A variety of 4-(1-alkenyl)-3-arylisoquinolines have been prepared in moderate to excellent yields by the Pd(II)-catalyzed cyclization of 2-(1 alkynyl)arylaldimines in the presence of various alkenes. The introduction of an o-methoxy group on the arylaldimine promotes the Pd-catalyzed cyclization and stabilizes the resulting Pd(II) intermediate, improving the yields of the isoquinoline products. Ketone-containing isoquinolines 36 and 49-51 have also been prepared by this process when unsaturated alcohols are employed as the alkenes. PMID- 12558425 TI - Efficient syntheses of 2-chloro-2'-deoxyadenosine (cladribine) from 2' deoxyguanosine(1). AB - We report efficient syntheses of the clinical agent cladribine (2-chloro-2' deoxyadenosine, CldAdo), which is the drug of choice against hairy-cell leukemia and other neoplasms, from 2'-deoxyguanosine. Treatment of 3',5'-di-O-acetyl- or benzoyl-2'-deoxyguanosine (1) with 2,4,6-triisopropyl- or 4-methylbenzenesulfonyl chloride gave high yields of the 6-O-arylsulfonyl derivatives 2 or 2'b. Deoxychlorination at C6 of 1 also proceeded to give the 2-amino-6-chloropurine derivative 5 in excellent yields. The nonaqueous diazotization/chloro dediazoniation (acetyl chloride/benzyltriethylammonium nitrite) of 2, 2'b, and 5 gave the 2-chloropurine derivatives 3, 3'b, and 6, respectively. The selective ammonolysis at C6 (arylsulfonate with 3 or chloride with 6) and accompanying deprotection of the sugar moiety gave CldAdo (64-75% overall yield from 1). PMID- 12558426 TI - Sulfur ylide-initiated thio-claisen rearrangements. The synthesis of highly substituted indolines. AB - The coupling of rhodium carbenoids from vinyl diazoacetates with 2-thio-3-alkyl indoles was found to generate C(3) quaternary substituted indolines via a thionium ylide-initiated [3,3]-sigmatropic rearrangement. PMID- 12558427 TI - 3-Bromopropenyl esters in organic synthesis: indium- and zinc-mediated entries to alk-1-ene-3,4-diols. AB - Metallic indium and zinc readily add to 3-bromopropenyl acetate (5) and benzoate (6) either in THF or in water, affording the corresponding 3-acyloxyallyl organometallic compounds. Nucleophilic addition to aldehydes opens a route to alk 1-ene-3,4-diols 2 in good to excellent yields. Two synthetic protocols were developed, the former involving indium in THF under Grignard conditions and the latter involving zinc in aqueous ammonium chloride under Barbier conditions. The diastereoselectivity, under all the conditions examined, mainly depends on the nature of the carbonyl compound; conjugated aldehydes afford syn adducts 2, while unconjugated aldehydes display the opposite anti stereopreference. PMID- 12558428 TI - Spin delocalization by triple-bonded functionalities in propargyl and heteropropargyl radicals, assessed from the EPR-spectral D parameter of 1,3 cyclopentanediyl triplet diradicals. AB - The cyclopentane-1,3-diyl triplet diradicals T and T' with the triplet-bonded acetylene, cyano, and isocyano functionalities at one of the radical sites are readily prepared from the corresponding azoalkanes by photodenitrogenation in a 2 methyltetrahydrofuran (2-MTHF) matrix at 77 K. The EPR-spectral D values of these triplet diradicals show that the spin delocalizing ability of the triple-bonded pi substituent follows the order -Ctbd1;CH > -NC approximately -CN. Good correlations of the D values have been obtained with the hyperfine coupling constants (a(H)) and with the calculated spin densities (PM3/AUHF-CI method) of the corresponding monoradicals M. The propargyl-type mesomeric structure is favored over the allenyl-type contributor for all three triple-bonded functionalities; spin delocalization is less pronounced in the heteropropargyl derivatives due to the electronegativity effect of the nitrogen atom. PMID- 12558429 TI - Intramolecular hydrogen abstraction reaction promoted by N-radicals in carbohydrates. Synthesis of chiral 7-oxa-2-azabicyclo[2.2.1]heptane and 8-oxa-6 azabicyclo[3.2.1]octane ring systems. AB - The reaction of phenyl and benzyl amidophosphates and alkyl and benzyl carbamate derivatives of aminoalditols with (diacetoxyiodo)benzene or iodosylbenzene and iodine is a mild and selective procedure for the synthesis of chiral 7-oxa-2 azabicyclo[2.2.1]heptane and 8-oxa-6-azabicyclo[3.2.1]octane ring systems under neutral conditions. This reaction can be considered to be an intramolecular N glycosidation that goes through an intramolecular 1,5-hydrogen abstraction promoted by an N-amido radical followed by oxidation of the transient C-radical intermediate to an oxycarbenium ion. This methodology proved to be useful not only as a suitable strategy for the preparation of these bicyclic arrays but also for the selective oxidation of specific carbons of the carbohydrate skeleton, constituting a good procedure for the synthesis of protected N,O-uloses. PMID- 12558430 TI - Preparation and chemistry of phosphoranyl-derived iodanes. AB - The preparation and chemistry of novel phosphoranyl-derived lambda(3)-iodanes is reported. The phosphoranyl-derived phenyliodonium sulfonates were prepared in good yields by the reaction of stabilized phosphonium ylides [1 triphenylphosphoranylidene-2-propanone, methyl(triphenylphosphoranylidene)acetate, (triphenylphosphoranylidene)acetaldehyde, and (triphenylphosphoranylidene)acetonitrile] with the pyridinium complex of iodobenzene ditriflate or with [hydroxy(tosyloxy)iodo]benzene under mild conditions. These compounds represent a potentially useful class of reagents that combine in one molecule synthetic advantages of a phosphonium ylide and an iodonium salt. Specifically, phosphorane-derived phenyliodonium tosylates can react with soft nucleophiles, such as iodide, bromide, benzenesulfinate, and thiophenolate anions, with a selective formation of the respective alpha functionalized phosphonium ylides, which can be further converted to alkenes by the Wittig reaction with benzaldehyde. The phosphoranyl-derived benziodoxoles can be prepared by the reaction of 1-acetoxybenziodoxole with stabilized phosphonium ylides. An unusual ligand exchange on the iodine(III) center resulting in the substitution of a carbon ligand with an oxygen ligand was observed in the reaction of these compounds with strong acids. PMID- 12558431 TI - Electroreductive intramolecular cyclization of a bromo propargyloxy ester catalyzed by nickel(i) tetramethylcyclam electrogenerated at carbon cathodes in dimethylformamide. AB - Cyclic voltammetry and controlled-potential electrolysis have been employed to investigate and characterize the reductive intramolecular cyclization of ethyl 2 bromo-3-(3',4'-dimethoxyphenyl)-3-(propargyloxy)propanoate (1) promoted by (1,4,8,11-tetramethyl-1,4,8,11-tetraazacyclotetradecane)nickel(I), [Ni(tmc)](+), electrogenerated at glassy carbon cathodes in dimethylformamide containing tetraalkylammonium salts. Cyclic voltammograms for reduction of [Ni(tmc)](2+) in the presence of 1 reveal that [Ni(tmc)](+) catalytically reduces 1 at potentials more positive than those required for direct reduction of 1. During controlled potential electrolyses of solutions containing [Ni(tmc)](2+) and 1, catalytic reduction of the latter proceeds via one-electron cleavage of the carbon-bromine bond to form a radical intermediate that undergoes cyclization to afford 2-(3',4' dimethoxyphenyl)-3-(ethoxycarbonyl)-4-methylenetetrahydrofuran (2). In the presence of a base (either electrogenerated or deliberately added as potassium tert-butoxide), 2 rearranges to give 2-(3',4'-dimethoxyphenyl)-3-(ethoxycarbonyl) 4-methyl-2,5-dihydrofuran (3). A mechanistic scheme is proposed to explain the results obtained by means of cyclic voltammetry and controlled-potential electrolysis. PMID- 12558432 TI - Formal synthesis of (+/-)-guanacastepene A. AB - A 17 step synthesis of 55, a late intermediate in Danishefsky's guanacastepene A synthesis, has been completed in 4% overall yield. Key features include the use of vinylmagnesium bromide in the Pd-catalyzed coupling with triflate 13 to give triene 16 without the formation of Heck products, a novel extension of the Stork Jung vinylsilane Robinson annulation that provides tricyclic 2 hydroxymethylcyclohexenone 42 from 23b in four steps and 51% yield, the ability to obtain almost exclusively alpha'-alkylation of 35ba by the proper choice of protecting groups, and the ability to obtain the desired beta-alcohol selectively by reduction of keto alcohol 42 rather than keto ester 53. PMID- 12558433 TI - Design and synthesis of novel fluoropeptidomimetics as potential mimics of the transition state during peptide hydrolysis. AB - alpha-Fluoroamino acids were targeted in our ongoing efforts to design novel fluoropeptidomimetics (1) as potential protease inhibitors. alpha-Fluoroglycine derivative (2) and alpha-fluoro-beta-aminoethanethiol derivatives (3-9) were synthesized for the first time en route to obtain the peptidomimetic moiety 1. The stability of 2-9 was investigated under organic as well as aqueous conditions. The stability of 3-9 under acidic and basic conditions, the effect of substitution at C-2 position, and potential biological activities are discussed. PMID- 12558434 TI - Ion channels from linear and branched bola-amphiphiles. AB - The synthesis and characterization of the ion channel activity of three new bola amphiphiles is described. These compounds are conceptually derived from a previously reported bis-cyclophane bola-amphiphile through opening of the cyclophanes to acyclic structures and were found to readily form ion channels in planar bilayer membranes as assessed by bilayer clamp single-channel analysis. All three compounds behaved very similarly: the dominant channels formed by all three are Ohmic with specific conductance of 10 +/- 1 pS (NaCl electrolyte) and 39 +/- 1 pS (CsCl electrolyte). Single-ion permeability ratios, determined from dissymmetric electrolyte experiments, showed the selectivity P(Cs(+)) > P(Na(+)) > P(Cl(-)). Less frequently, lower conductance channels were also observed to act independently of the dominant channels. The lifetimes of the dominant channels range from 70 to 280 ms for the three compounds with some very long-lived openings (20-40 s) observed for two of the three. The lower conductance states have shorter lifetimes. This study demonstrates that bis-macrocyclic compounds are not essential for channel formation by bola-amphiphiles, and opens a new class of channel-forming compounds for structure-activity optimization. PMID- 12558435 TI - Construction of monomeric and polymeric porphyrin compartments by a Pd(II) pyridine interaction and their chiral twisting by a BINAP ligand. AB - The construction of chirally twisted porphyrin-based molecular capsule 6 and polymeric capsule 8 was investigated by means of scanning electron microscopy (SEM) and (1)H NMR, UV-visible, and CD spectroscopic observations. Molecular capsule 6 and polymeric capsule 8 were constructed by the reaction of chiral cis Pd(II) complex 4 bearing a (R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP) ligand with porphyrin 1 bearing four pyridyl groups and porphyrin 2 bearing eight pyridyl groups, respectively. The peak-splitting pattern of the beta-pyrrole protons in the (1)H NMR spectrum and the specific CD spectral pattern bearing an exciton coupling band indicate that both molecular capsule 6 and polymeric capsule 8 are chirally twisted. Moreover, it was found that the CD intensity of the polymeric capsule plotted against [4]/([4] + [3]) shows a sigmoidal curvature, reflecting a unique cooperativity among the ligand groups; that is, the ligand existing in excess over the other dominates the twisting direction. These results consistently demonstrate that "chirality" in these molecular assembly systems is conveniently controlled by the use of chiral ligands. PMID- 12558436 TI - Phenonium ions from the addition of phenyl cations to alkenes. Photochemical synthesis of (rearranged) aminoalkylanilines from haloanilines in the presence of alkenes and amines. AB - beta-Aminoalkylanilines are smoothly obtained by irradiation of 4-chloro- and 4 fluoroanilines (as well as the N,N-dimethyl derivatives) in the presence of alkenes (1-hexene, cyclohexene) and amines (butylamine, piperidine) in polar, protic solvents such as trifluoroethanol (yield 40-75%). The reaction involves photoheterolysis of the haloaniline, addition of the resulting phenyl cation to the alkene and trapping of the phenonium cation by amine. A fraction (up to ca. 20%) of aminoalkylanilines resulting from Wagner-Meerwein rearrangement of the phenonium cation is obtained in some cases. Reduction and direct trapping of the phenyl cation by the amine compete with the above three-component synthesis in a less stabilizing solvent such as acetonitrile, but not in CF(3)CH(2)OH. PMID- 12558437 TI - Investigation of bicyclic thioketones as triggers for liquid crystal optical switches. AB - The axially chiral bicyclic thioketones 11 and 15 were prepared and investigated for suitability as chiroptical triggers in a liquid crystal optical switch. Irradiation of partially resolved 15 with unpolarized light leads to its conversion to the racemic form (photoracemization). However, irradiation of racemic thioketones 11 and 15 with circularly polarized light does not lead to detectable photoresolution. The lack of photoresolution was traced to inefficiency in intramolecular, through-bond triplet energy transfer. These thioketones are not suitable for use as phototriggers. PMID- 12558438 TI - (2,5)-Ene cyclization catalyzed by mesoporous solid acids: isotope labeling study and ab initio calculation for continuum from concerted to stepwise ene mechanism. AB - (2,5)-Ene reactions catalyzed by mesoporous solid acids are reported from the mechanistic point of view. The continuum (2,5)-ene mechanism from the concerted to the cationic cyclization followed by 1,2-hydride shift is evaluated. The solid acid-catalyzed cyclization of the oxonium ion intermediate 4 derived from cyclic allylic lactol ether 3 bearing allylic hydroxy group affords the (2,5)-ene product as the enol form, eventually tautomerizing to the corresponding aldehyde 6. The continuum from the concerted to stepwise mechanism is experimentally and theoretically verified in the present ene cyclization of the oxonium ion intermediate such as 4. The stepwise cyclization leading to aldehyde 6 is thus shown to associate with the concerted version as a result of the stabilization of the beta-hydroxycarbenium ion intermediate. PMID- 12558439 TI - Contracted and expanded meso-alkynyl porphyrinoids: from triphyrin to hexaphyrin. AB - The boron trifluoride-catalyzed Rothemund condensation of triisopropylsilyl (TIPS) propynal 1 with 3,4-diethylpyrrole in dichloromethane, followed by oxidation with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) generates a mixture of products, including [15]triphyrin(1.1.3) H3, corrole H(3)4, porphyrin H(2)2, [24]pentaphyrin(1.1.1.1.1) H(4)5, [28]hexaphyrin(1.1.1.1.1.1) H(4)6, and two linear tripyrromethenes H(2)7 and H(2)8. We report the spectroscopic characteristics of these unusual chromophores, together with the crystal structures of triphyrin H3 (and its zinc complex ZnCl3), porphyrin H(2)2 (and its metal complexes Zn2, Ni2 and Pt2), hexaphyrin H(4)6, and tripyrromethene nickel(II) complex Ni7. When the condensation is catalyzed with trifluoroacetic acid, rather than boron trifluoride, the triphyrin H3 become the main product (26% yield). This novel macrocycle is linked with a TIPS-substituted exocyclic double bond. This C=C bond makes an eta(2)-interaction with the zinc center in ZnCl3 with C-Zn distances of 2.863 and 3.025 A. The porphyrin H(2)2 is severely ruffled, and its absorption spectrum is red-shifted and broadened compared with the analogous compound without ethyl substituents. The hexaphyrin H(4)6 adopts a figure-of-eight conformation with virtual C(2) symmetry in the solid state and C(2) symmetry in solution on the NMR time scale. Oxidation with DDQ appears to convert this nonaromatic [28]hexaphyrin into an aromatic [26]hexaphyrin with a strongly red-shifted absorption spectrum, but the oxidized macrocyle is too unstable to isolate. PMID- 12558440 TI - A novel type of hydrogen-bonded assemblies based on the melamine.cyanuric acid motif. AB - This paper reports the formation of novel hydrogen-bonded assemblies 1(3).CA obtained upon mixing cyanuric acid (CA) with melamine derivatives 1, in which two of the three possible H-bonding arrays have been blocked. The four components are held together by 9 hydrogen bonds and form a rigid planar structure in which a central CA (three ADA motifs: A = acceptor, D = donor) is hydrogen bonded to three peripheral melamine derivatives (DAD motif). Furthermore, the synthesis and assembly studies are described of hydrogen-bonded assemblies 2-4.CA, comprised of three melamine derivatives that are covalently connected, and CA. The overall thermodynamic stability of assemblies 2-4.CA is superior to 1(3).CA (I(Tm) = 9 vs 3.6). The presence of the 2.CA complex in chloroform was confirmed by (1)H NMR spectroscopy and MALDI-TOF mass spectrometry. Substitution of the trimelamines with chiral or fluorescent groups (R(3)) enabled the study of the assemblies by CD and fluorescence spectroscopy. Titration experiments revealed strongly enhanced stabilities even in the presence of polar solvents, such as THF and CH(3)OH. Depending on the polarity of the solvent, stacking between the planar assembly units was observed. PMID- 12558441 TI - Enantioselective [6pi]-photocyclization reaction of an acrylanilide mediated by a chiral host. Interplay between enantioselective ring closure and enantioselective protonation. AB - The [6pi]-photocyclization of the anilides 1a and 5 was studied in the absence and in the presence of the enantiomerically pure chiral lactam 4. The relative configuration of the products was unambiguously established by single-crystal X ray crystallography and by NMR spectroscopy. A significant enantiomeric excess was observed upon reaction of compound 1a to its photocyclization products at -55 degrees C employing lactam 4 as a chiral complexing agent in toluene as the solvent (66% yield). The trans product ent-3a was obtained in 57% ee, and the minor diastereoisomer (trans/cis = 73/27), cis product ent-2a, was obtained in 30% ee. DFT calculations were conducted modeling the complexation of intermediates 8 and ent-8 to host 4. In agreement with steric arguments concerning the conrotatory ring closure of 1a, the formation of ent-8 is favored leading to the more stable complex 4.ent-8 as compared to 4.8. Whereas the enantioselectivity in the photocyclization to trans compound ent-3a increased upon reduction in the reaction temperature, the enantiomeric excess in the formation of cis compound ent-2a went through a maximum at -15 degrees C (45% ee) and decreased at lower temperatures. Deuteration experiments conducted with the pentadeuterated analogue of 1a, d(5)-1a, revealed that the protonation of the intermediates 8 and ent-8 is influenced by chiral amide 4. In the formation of ent-3a/3a, both the enantioselective ring closure and the enantioselective protonation by amide 4 favor the observed (6aS,10aS)-configuration of the major enantiomer ent-3a. In the formation of ent-2a/2a, the enantioselective ring closure (and the subsequent diastereoselective protonation) favors the (6aR,10aS) configuration that is found in compound 2a. Contrary to that, the enantioselective protonation by amide 4 shows a preference for ent-2a with the (6aS,10aR)-configuration. PMID- 12558442 TI - Carbocation-forming reactions in dimethyl sulfoxide. AB - Mesylate derivatives of 3-aryl-3-hydroxy-beta-lactams and thiolactams react in DMSO-d(6) by first-order processes to give alcohol products. Substituent effect studies implicate carbocation intermediates (ion-pairs) that are captured by DMSO d(6) to give transient oxosulfonium ions. Rapid reaction of the oxosulfonium ions with trace amounts of water leads to the alcohol product and regenerates DMSO d(6). H(2)(17)O labeling studies show that (17)O is incorporated into the DMSO. The mesylate derivatives of endo- and exo-2-hydroxy-2-phenylbicyclo[2.2.1]heptan 3-one also react in DMSO-d(6) to give the alcohol products. Ion-pair intermediates that capture DMSO giving unstable oxosulfonium ions are again proposed. Exo-2-phenyl-endo-bicyclo[2.2.1]heptyl trifluoroacetate readily eliminates trifluoroacetic acid in DMSO-d(6) via a cationic mechanism involving loss of the endo-trifluoroacetate leaving group as well as an exo-hydrogen. The O methyl oxime derivative of alpha-chloro-alpha,alpha-diphenylacetophenone reacts in DMSO-d(6) to give 1-methoxy-2,3-diphenylindole, a product derived from cyclization of a cationic intermediate. A common ion rate suppression provides further evidence for a cationic mechanism. The triflate derivative of pivaloin reacts by a cationic mechanism in DMSO-d(6) to give rearranged products. The rate is even faster than in highly ionizing solvents such as trifluoroethanol or trifluoroacetic acid. 1-Adamantyl mesylate reacts in DMSO-d(6) by a first-order process (Y(OMs) = -4.00) to give a long-lived oxosulfonium ion, 1-Ad OS(CD(3))(2)(+), which can be characterized spectroscopically. This oxosulfonium ion reacts only slowly with water at elevated temperatures to give 1-adamantanol. DMSO is therefore a viable solvent for k(s), k(C), and k(Delta) cationic processes. PMID- 12558444 TI - Sequential elimination-cyclopropanation reactions promoted by samarium: highly diastereoselective synthesis of cyclopropylamides. AB - trans-Cyclopropanamides were obtained, in high yield, from 2-chloro-3 hydroxyamides by a sequenced elimination-cyclopropanation process promoted by Samarium/diiodomethane or Samarium diiodide and Samarium/diiodomethane. PMID- 12558443 TI - Short synthesis of tert-butyl-hydroxylated 3,5-di-tert-butyl-4 hydroxybenzaldehyde: Synthesis of tert-butyl-hydroxylated S-2474. AB - We have developed a very short synthesis of tert-butyl-hydroxylated di-tert-butyl 4-hydroxybenzaldehyde in which the HBr-DMSO system is used as an effective oxidant (overall yield of 45% for the entire four-step process from 2-tert-butyl p-cresol). We also accomplished the synthesis of a major metabolite of the antiarthritic drug candidate S-2474. PMID- 12558445 TI - Highly efficient recognition of native TpT by artificial ditopic hydrogen-bonding receptors possessing a conformationally well-defined linkage. AB - Synthesis and binding affinity of rationally designed artificial ditopic nucleobase receptors are reported. The ditopic receptors were designed to recognize thymine-thymine dinucleotides by their two hydrogen-bonding moieties, which are connected to conformationally well-defined linkages such as ferrocene and biphenylene. The ditopic receptors exhibited a remarkably strong binding affinity for lipophilic TpT analogue in CDCl(3)/DMSO-d(6) (85:15, v/v). The binding affinity of the ditopic receptors for the dinucleotide was so high that even native TpT was extracted by them into CDCl(3). Detailed comparisons for the recognition abilities of the ditopic receptors were also conducted. PMID- 12558446 TI - Bis(o-nitrophenyl)ethanediol: A practical photolabile protecting group for ketones and aldehydes. AB - The ketals of bis(o-nitrophenyl)ethanediol and ketones or aldehydes are smoothly deprotected in neutral conditions by irradiation with 350 nm light. The chemical stability in basic, acidic, and oxidizing media makes this form of protection orthogonal to classical protecting groups. Both racemic and enantiopure forms are readily available in two steps from inexpensive starting materials. PMID- 12558447 TI - Cobalt-catalyzed direct electrochemical cross-coupling between aryl or heteroaryl halides and allylic acetates or carbonates. AB - The electroreduction of a mixture of functionalized aromatic or heteroaromatic bromides or chlorides and allylic compounds such as acetates or carbonates in an electrochemical cell fitted with a sacrificial iron anode affords, in the presence of cobalt halide associated with pyridine as ligand in acetonitrile or DMF, the corresponding coupling product in good yields. PMID- 12558448 TI - A mild deprotection strategy for allyl-protecting groups and its implications in sequence specific dendrimer synthesis. AB - A mild deprotection strategy for allyl ethers under basic conditions in the presence of a palladium catalyst is described. Under these conditions, aryl allyl ethers can be cleaved selectively in the presence of alkyl allyl ethers. These conditions are also effective in the deprotection of allyloxycarbonyl groups. The utility of the current methodology in sequence specific dendrimer synthesis is demonstrated. PMID- 12558449 TI - A practical synthesis of the F-ring of halichondrin B via ozonolytic desymmetrization of a C(2)-symmetric dihydroxycyclohexene. AB - C(2)-symmetric dihydroxycyclohexene 1 was desymmetrized via a one-pot Criegee ozonolysis/acylation protocol to afford acetal-lactone 2. Installation of the allyl side chain on the convex face of the bicyclic system and subsequent reduction provided the desired tetrahydrofuran 4 with the correct relative and absolute stereochemistries. Simple functional group manipulations led to the desired F-ring module 3 of halichondrin B. PMID- 12558450 TI - Efficient synthesis of novel 1alpha-amino and 3beta-amino analogues of 1alpha,25 dihydroxyvitamin d(3). AB - Convenient synthetic routes to 1alpha-amino-25-hydroxyvitamin D(3) (3) and 3beta amino-3-deoxy-1alpha,25-dihydroxyvitamin D(3) (4), novel analogues of vitamin D(3) bearing an amino group at the C-1 or C-3 position, have been developed starting from (S)-(+)-carvone. Construction of the A-ring fragments was accomplished by selective enzymatic hydrolysis of a diester intermediate and introduction of the amino group under Mitsunobu conditions. PMID- 12558451 TI - Direct conversion of aldehydes to amides, tetrazoles, and triazines in aqueous media by one-pot tandem reactions. AB - A variety of aldehydes reacted with iodine in ammonia water at room temperature to give the nitrile intermediates, which were trapped by addition of hydrogen peroxide, sodium azide, or dicyandiamide to produce their corresponding amides, tetrazoles, and 1,3,5-triazines in modest to high yields. The one-pot tandem reactions were conducted in water media, and the products were obtained simply by extraction or filtration. PMID- 12558452 TI - Use of trichloroacetimidate linker in solid-phase Peptide synthesis. AB - A solid-phase method for the preparation of C-terminal amino-alcohol-containing peptides using activated Wang resin is presented. A diverse set of (fluorenylmethoxy)carbonyl (Fmoc) protected amino alcohols was found to load rapidly and efficiently. The synthetic utility of this approach was demonstrated through the direct synthesis of the peptide drug octreotide with excellent yield and purity. These results suggest that the use of trichloroacetimidate activated resins offers an attractive alternative in the preparation of this class of peptides. PMID- 12558453 TI - Palladium-catalyzed amination of aryl bromides with hindered N-alkyl-substituted anilines using a palladium(I) tri-tert-butylphosphine bromide dimer. AB - An efficient palladium-catalyzed amination of aromatic bromides with hindered N alkyl-substituted anilines is described, either using the combination of Pd(OAc)(2) and P(t-Bu)(3) or a palladium(I) tri-tert-butylphosphine bromide dimer, [Pd(mu-Br)(t-Bu(3)P)](2), a new, commercially available, and easily handled catalyst. PMID- 12558454 TI - A highly convenient, efficient, and selective process for preparation of esters and amides from carboxylic acids using Fe(3+)-K-10 montmorillonite clay. AB - In the presence of Fe(3+)-K-10 montmorillonite clay as a catalyst, aliphatic carboxylic acids selectively produced the corresponding esters in the presence of aromatic carboxylic acids by treatment with alcohols. Both the aliphatic and aromatic carboxylic acids formed the amides by reacting with the aliphatic amines, but only the aliphatic carboxylic acids yielded the anilides by treatment with aromatic amines. The catalyst is recoverable and recyclable. PMID- 12558455 TI - Silaaromaticity in polycyclic systems: a computational study. AB - Density functional theory (B3LYP) calculations were performed to examine the effect of Si substitution on the aromaticity of some polycyclic hydrocarbons using geometric criterion (HOMA), isodesmic isomerization reactions, homodesmotic equations, NICS values, chemical hardness, and out-of-plane distortive tendencies. The HOMA values are lower and the NICS values are higher in the Si substituted rings compared to those in the hydrocarbon counterpart, whereas the homodesmotic equations predict little loss of aromaticity upon Si replacement in polycylic systems. The chemical hardness values decrease and the out-of-plane distortive tendency increases upon silicon substitution. The relative energies of the positional isomers and the causative factors are analyzed. The high reactivity of some silaaromatics toward dimerization is explained based on local softness indices. PMID- 12558456 TI - An expeditious and efficient procedure for the synthesis of unsaturated acyclonucleosides of Z configuration related to D4T. AB - Enantiopure 2,5-dihydrofuran derivatives were prepared from (S)-glycidol through a new reaction sequence involving epoxide opening with a vinylcuprate, selenium induced cyclization to give exclusively the 5-endo product, and regioselective selenoxide elimination. Unsaturated acyclonucleosides of Z configuration were obtained in a straightforward manner by treating 2,5-dihydrofuran with iodotrimethylsilane in the presence of silylated purinic or pyrimidinic bases. This synthetic process involves opening of the dihydrofuran ring by trimethylsilyl iodide and substitution of iodine by the nucleic base in a single reaction step. PMID- 12558457 TI - Vaccines against drugs of abuse: a viable treatment option? AB - Drug addiction is a chronically relapsing brain disorder. There is an urgent need for new treatment options for this disease because the relapse rate among drug abusers seeking treatment is quite high. During the past decade, many groups have explored the feasibility of using vaccines directed against drugs of abuse as a means of eliminating illicit drug use as well as drug overdose and neurotoxicity. Vaccines work by inducing drug-specific antibodies in the bloodstream that bind to the drug of abuse and prevent its entry into the brain. The majority of work in this area has been conducted with vaccines and antibodies directed against cocaine and nicotine. On the basis of preclinical work, vaccines for cocaine and nicotine are now in clinical trials because they can offer long-term protection with minimal treatment compliance. In addition, vaccines and antibodies for phencyclidine, methamphetamine and heroin abuse are currently under development. An underlying theme in this research is the need for high concentrations of circulating drug-specific antibodies to reduce drug-seeking and drug-taking behaviour when the drug is repeatedly available, especially in high doses. Although vaccines against drugs of abuse may become a viable treatment option, there are several drawbacks that need to be considered. These include: a lack of protection against a structurally dissimilar drug that produces the same effects as the drug of choice;a lack of an effect on drug craving that predisposes an addict to relapse; and tremendous individual variability in antibody formation. Forced or coerced vaccination is not likely to work from a scientific perspective, and also carries serious legal and ethical concerns. All things considered, vaccination against a drug of abuse is likely to work best with individuals who are highly motivated to quit using drugs altogether and as part of a comprehensive treatment programme. As such, the medical treatment of drug abuse will not be radically different from treatment of other chronic diseases. PMID- 12558458 TI - Extended spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae: considerations for diagnosis, prevention and drug treatment. AB - Extended spectrum beta-lactamase (ESBL)-producing organisms pose unique challenges to clinical microbiologists, clinicians, infection control professionals and antibacterial-discovery scientists. ESBLs are enzymes capable of hydrolysing penicillins, broad-spectrum cephalosporins and monobactams, and are generally derived from TEM and SHV-type enzymes. ESBLs are often located on plasmids that are transferable from strain to strain and between bacterial species. Although the prevalence of ESBLs is not known, it is clearly increasing, and in many parts of the world 10-40% of strains of Escherichia coli and Klebsiella pneumoniae express ESBLs. ESBL-producing Enterobacteriaceae have been responsible for numerous outbreaks of infection throughout the world and pose challenging infection control issues. Clinical outcomes data indicate that ESBLs are clinically significant and, when detected, indicate the need for the use of appropriate antibacterial agents. Unfortunately, the laboratory detection of ESBLs can be complex and, at times, misleading. Antibacterial choice is often complicated by multi-resistance. Many ESBL-producing organisms also express AmpC beta-lactamases and may be co-transferred with plasmids mediating aminoglycoside resistance. In addition, there is an increasing association between ESBL production and fluoroquinolone resistance. Although in in vitro tests ESBLs are inhibited by beta-lactamase inhibitors such as clavulanic acid, the activity of beta-lactam/beta-lactamase inhibitor combination agents is influenced by the bacterial inoculum, dose administration regimen and specific type of ESBL present. Currently, carbapenems are regarded as the drugs of choice for treatment of infections caused by ESBL-producing organisms. Unfortunately, use of carbapenems has been associated with the emergence of carbapenem-resistant bacterial species such as Stenotrophomonas sp. or Pseudomonas sp. PMID- 12558459 TI - Interactions between cyclosporin and lipid-lowering drugs: implications for organ transplant recipients. AB - Dyslipidaemia is more frequent in solid organ transplant recipients than in the general population, primarily as a result of immunosuppressive drug treatment. Both cyclosporin and corticosteroids are associated with dyslipidaemic adverse effects. In order to reduce the overall cardiovascular risk in these patients, lipid-lowering drugs have become widely used, especially HMG-CoA reductase inhibitors (statins). Cyclosporin, as well as most statins (lovastatin, simvastatin, atorvastatin and pravastatin) are metabolised by cytochrome P450 (CYP)3A4, so a bilateral pharmacokinetic interaction between these drugs is theoretically possible. However, results from several studies show that statins do not induce increased systemic exposure of cyclosporin. A small (but not clinically relevant) reduction in systemic exposure of cyclosporin has actually been shown in many studies. Cyclosporin-treated patients on the other hand show several-fold higher systemic exposure of all statins, both those that are metabolised by CYP3A4 and fluvastatin (metabolised by CYP2C9). Therefore, the mechanism for this interaction does not seem to be solely caused by inhibition of CYP3A4 metabolism, but it is probably also a result of inhibition of statin transport in the liver, at least in part. Other lipid-lowering drugs, such as fibric acid derivatives, bile acid sequestrants, probucol, fish oils and orlistat are also used in solid organ transplant recipients. Most of them do not interact with cyclosporin, but there are reports indicating that both probucol and orlistat may reduce cyclosporin bioavailablility to a clinically relevant degree. There is no information on possible interaction effects of cyclosporin on the pharmacokinetics of lipid-lowering drugs other than statins, but it is not likely that any clinical relevant interference exists with fish oil, orlistat, probucol or bile acid sequestrants. PMID- 12558460 TI - Management of insomnia in patients with chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a common medical disorder, which causes considerable morbidity and mortality. Given the chronic and symptomatic nature of the disease, the patient is often seen in the physician's office with complaints of dyspnea. However, more than 50% of COPD patients also have sleep complaints characterised by longer latency to falling asleep, more frequent arousals and awakenings, and/or generalised insomnia. Sleep disturbance tends to be more severe with advancing disease and substantially reduces the COPD patients' quality of life. In approaching the COPD patient who complains of insomnia it is important to take a complete sleep history. Having characterised the degree and duration of the problem, medical management of the underlying COPD must first optimise oxygen saturation while minimising the effects of many of the medications used for COPD. While aerosol therapies may be systemically absorbed and contribute to sleep disruption, anticholinergics, such as ipratropium bromide, are the least likely to do so and indeed have been shown to improve sleep quality in this population. Many of the traditional sedatives and hypnotics have been used in the COPD population including benzodiazepines, imidazopyridines, pyrazolopyrimidines and, less commonly, antidepressants and phenothiazines. Clinical trials support the role of numerous agents in treating insomnia in this population but do not always provide reassurance that these therapies can be used safely, particularly in the patient with severe COPD with hypercarbia. Benzodiazepines are among the most commonly employed agents, but case reports and series continue to describe adverse pulmonary events. Although the newer pyridine derivatives also have the potential to worsen pulmonary function, they appear less likely to do so. Data to date are limited with the tricyclic antidepressants and phenothiazines, although they appear to be very well tolerated from a respiratory point of view. Since sleep disturbances are often long-standing and associated with maladaptive behaviours towards sleep, cognitive/behavioural approaches are often useful and are more effective in the long-term than are hypnotics. When prescription of a sedative is to be made, extra caution is required for those patients at increased risk of adverse respiratory effects, such as those with advanced disease and hypercarbia in whom pharmacological therapy is often best avoided. Selection of the various options will depend upon the degree of underlying disease and the patient's specific complaints of insomnia. Finally, it is important to remember that while most hypnotics work in an acute setting, the long-term management will require an integrated approach. PMID- 12558462 TI - Losartan in diabetic nephropathy. AB - Losartan is an orally active, selective, nonpeptide, angiotensin II AT(1) receptor antagonist. Losartan 50 or 100 mg/day was significantly more effective than placebo in reducing the incidence of a doubling of serum creatinine, end stage renal disease (ESRD) or death (43.5% vs 47.1%, p = 0.02) in a pivotal, well designed trial (Reduction of Endpoints in Non insulin dependent diabetes mellitus with the Angiotensin II Antagonist Losartan [RENAAL] study) in 1513 patients with type 2 diabetes mellitus and proteinuria. Losartan also significantly reduced the incidence of doubling of serum creatinine level (p = 0.006), ESRD (p = 0.002), ESRD or death (p = 0.01) and doubling of serum creatinine and ESRD (p = 0.01) compared with placebo in the RENAAL trial. There were similar incidences of overall mortality and morbidity and mortality from cardiovascular causes between treatment groups. In addition, data from several nonblind and double-blind studies indicates that losartan effectively reduces the mean albumin excretion rate. Two double-blind studies show that losartan has similar effects to enalapril on kidney function. Data from 4058 patients (3300 with essential hypertension) who have received losartan (10-150 mg/day) in clinical trials indicate it is well tolerated. In the RENAAL study 17.2% and 21.7% of losartan and placebo recipients discontinued treatment because of adverse events, but causality was not determined. PMID- 12558466 TI - NICE methodological guidelines and decision making in the National Health Service in England and Wales. AB - The National Institute for Clinical Excellence (NICE) responds to requests by the Department of Health for guidance on the use of selected new and established technologies in the National Health Service (NHS) in England and Wales. This paper asks whether the NICE methodological guidelines help NHS decision makers meet the objectives of maximum health improvements from NHS resources and an equitable availability of technologies. The analytical basis of the guidelines is a comparison of the costs and consequences of new and existing methods of dealing with particular conditions using the incremental cost-effectiveness ratio. We explain why information on the costs and consequences of a particular technology in isolation is insufficient to address issues of efficiency of resource use. We argue that to increase efficiency, decision makers need information on opportunity costs. We show that in the absence of such information decision makers cannot identify the efficient use of resources. Finally we argue that economics provides valid methods for identifying the maximisation of health improvements for a given allocation of resources and we describe an alternative practical approach to this problem. Drawing on the experience of Ontario, Canada where an approach similar to that proposed by NICE has been in use for almost a decade, and recent reports about the consequences of NICE decisions to date, we conclude that instead of increasing the efficiency or equity of the use of NHS resources, NICE methodological guidelines may lead to: uncontrolled increases in NHS expenditures without evidence of any increase in total health improvements; increased inequities in the availability of services; and concerns about the sustainability of public funding for new technologies. PMID- 12558465 TI - Zoledronic acid: a review of its use in the management of bone metastases and hypercalcaemia of malignancy. AB - Zoledronic acid (Zometa) is an effective inhibitor of osteoclast-mediated bone resorption. Zoledronic acid demonstrated efficacy in the reduction of skeletalrelated events (SREs) in patients with multiple myeloma or bone metastases secondary to breast cancer, prostate cancer or other solid tumours, or hypercalcaemia of malignancy. Zoledronic acid was effective in patients with multiple myeloma or metastatic breast cancer with osteolytic or mixed bone lesions. The proportion of patients who experienced an SRE was similar during 12 months of treatment with zoledronic acid 4mg or pamidronic acid 90mg, but significantly fewer patients receiving zoledronic acid required radiotherapy to bone. Furthermore, in patients with breast cancer and osteolytic lesions, median time to a first SRE was more than 4 months longer with zoledronic acid than with pamidronic acid. In the multiple event analysis in a 12-month extension study (total study duration was 25 months) in patients with breast cancer, zoledronic acid was superior to pamidronic acid, with an 18% reduction in the risk of experiencing an SRE. Both drugs were associated with a slight reduction in pain. Zoledronic acid 4mg, compared with placebo, significantly reduced the proportion of patients with prostate cancer bone metastases experiencing an SRE, particularly pathological fractures after 15 months' treatment. The drug also significantly delayed the onset of skeletal complications compared with placebo in patients with prostate cancer and other solid tumours including non-small cell lung cancer. When administered as a single 15-minute intravenous infusion, zoledronic acid 4mg was significantly more effective than pamidronic acid administered as a 2-hour infusion in the treatment of severe hypercalcaemia of malignancy, as assessed by complete responses measuring normalised serum calcium concentrations at day 10 after a single dose. Furthermore, zoledronic acid normalised serum calcium concentrations significantly faster than pamidronic acid, and the duration of response and median time to relapse were approximately twice as long in zoledronic acid recipients than in pamidronic acid recipients. Zoledronic acid is well tolerated and has a similar tolerability profile to pamidronic acid. The most commonly reported adverse events included flu-like symptoms (fever, arthralgias, myalgias and bone pain), fatigue, gastrointestinal reactions, anaemia, weakness, dyspnoea and oedema. CONCLUSION: In conjunction with antitumour therapy, zoledronic acid should be considered for routine use to reduce skeletal complications in patients with advanced malignancies involving bone. In patients with hypercalcaemia of malignancy, zoledronic acid is expected to become the treatment of choice. PMID- 12558461 TI - Antimicrobial peptides: current status and therapeutic potential. AB - Antimicrobial peptides (AMPs) are effector molecules of the innate immune system. A variety of AMPs have been isolated from species of all kingdoms and are classified based on their structure and amino acid motifs. AMPs have a broad antimicrobial spectrum and lyse microbial cells by interaction with biomembranes. Besides their direct antimicrobial function, they have multiple roles as mediators of inflammation with impact on epithelial and inflammatory cells influencing diverse processes such as cell proliferation, immune induction, wound healing, cytokine release, chemotaxis and protease-antiprotease balance. AMPs qualify as prototypes of innovative drugs that may be used as antimicrobials, anti-lipopolysaccharide drugs or modifiers of inflammation. Several strategies have been followed to identify lead candidates for drug development, to modify the peptides' structures, and to produce sufficient amounts for pre-clinical and clinical studies. This review summarises the current knowledge about the basic and applied biology of AMPs. PMID- 12558467 TI - Treatment of atopic dermatitis and impact on quality of life: a review with emphasis on topical non-corticosteroids. AB - Atopic dermatitis (AD) is a chronic skin disease with increasing prevalence and rising costs. Stigmatisation and pruritus are only some aspects of potential quality-of-life (QOL) impairments. AD is not curable and repeated treatments are often necessary. At present, treatment with topically-applied corticosteroids is state-of-the-art for mild to moderate flare-ups. However, many patients are worried about the use of corticosteroids due to the widespread fear of adverse effects. In this review the present literature is analysed concerning impact on quality of life for topically-applicable alternatives to the state-of-the-art treatment. For comparison reasons, data from other treatment modalities are additionally given. Characteristics of studies were analysed using 'general' (year and mode of publication, type and aim of study, number of patients, and clinical measurement) and 'QOL specific' criteria (type and number of QOL measurements including relevance for study aim and age group, validation in used language, sensitivity to change, and improvement at end of study). QOL data are published only in the minority of studies evaluating treatment efficacy and do not cover the variety of possible therapies. Data are available for tacrolimus, pimecrolimus, UVA/UVB combination and UVB narrowband (topical non corticosteroidal treatments), as well as for topical corticosteroids, cyclosporin, and inpatient treatment. All studies provided a marked improvement in quality of life after therapy. One study assessed quality of life after a treatment-free follow-up period obtaining a clear increase in impact on quality of life. Since studies used different QOL measurements and vary in inclusion criteria, treatment schedules and presentation of results, a comparison of QOL improvement is not recommended. A single randomised study compared topically applied non-corticosteroidal treatment (UVA/UVB combination) with another treatment modality (cyclosporin) and found no difference in QOL improvement. At present, there is a clear lack of controlled randomised studies evaluating different active treatment modalities and their impact on quality of life. Consensus meetings are desirable to formulate guidelines for the selection and correct use of QOL measurements. Patients' fear of side effects (e.g. concerning corticosteroids) should be integrated in QOL questionnaires for evaluation of possible compliance problems and real costs. Since relapse after treatment is frequent in AD, QOL measurements should also be performed after a treatment-free follow-up period. At present, we can not answer the question 'which treatment best improves quality of life in AD?'. PMID- 12558468 TI - The burden of age-related macular degeneration: results of a cohort study in two French referral centres. AB - OBJECTIVE: To describe the economic impact of age-related macular degeneration (AMD) and to assess its medical and non-medical costs. DESIGN AND SETTINGS: An observational study was carried out in 105 patients in two French centres in a sample of 105 French patients. All consecutive patients, consulting during a 3 week period, were included provided they were 60 years of age or older and they presented an exudative form of AMD with a distant visual acuity in the best eye < or = 20/40. Data collected included clinical items, treatment modalities, medical follow-up, transport costs, impact of AMD on living conditions and welfare payments related to visual impairment. Costs were presented in 2000 values. PERSPECTIVE: General payer perspective (Social Security, private health insurance and patient). RESULTS: Mean age was 79.3 years and ranged from 62.8-95 years. Average length of disease evolution was 3.5 years. During a 3-month period, patients had a mean of 2.6 visits to the ophthalmologist. Thirty percent of the patients used vascular medications and 72.4% had been previously treated by laser photocoagulation. Only 10% had benefited from visual rehabilitation. Annual AMD cost per patient was 3660.29 euros (EUR) [95% CI: 2881.92-4438.62]. Half of these annual costs were medical costs. Other major cost components were home help costs EUR904.91 [95% CI: 478.88-1330.94] and transport costs for care EUR542.73 [95% CI: 146.31-939.14]. Non-medical costs were significantly higher for patients with more severe disease. CONCLUSIONS: The economic argument that costs are higher in patients with the lowest visual acuity emphasises the necessity of early detection and treatment of patients with AMD. PMID- 12558469 TI - A meta-analysis of quality-of-life estimates for stroke. AB - BACKGROUND: Researchers performing cost-effectiveness analyses often incorporate quality-of-life (QOL) estimates. OBJECTIVE: To aid analysts, we performed a meta analysis to estimate quality of life for minor, moderate, and major stroke and assessed the relative importance of study design characteristics in predicting the quality of life of patients with stroke. METHODS: Through a systematic search we identified 20 articles reporting 53 unique QOL weights for stroke. Each article was read and QOL weights and study characteristics were recorded. We used a hierarchical linear model (HLM) to perform a meta-regression. The model included severity of stroke, elicitation method, respondents, and QOL scale bounds as explanatory variables. RESULTS: Severity of stroke (p < 0.0001) and the bounds of the scale (p = 0.0015) were significant predictors of quality of life, while the elicitation method and respondents were not. Pooling QOL weights using the HLM model, we estimated a quality of life of 0.52 for major stroke, 0.68 for moderate stroke, and 0.87 for minor stroke if the time trade-off method is used to assess quality of life from community members when the scale bounds range from death to perfect health. CONCLUSIONS: We found no systematic difference in stroke QOL weights depending on elicitation method or respondents. However, quality of life is sensitive to the bounds of the scale. Because the pooled QOL estimates reported here are based on a comprehensive review of the QOL literature for stroke, they should be of great use to researchers performing cost-utility analyses of interventions designed to prevent or treat stroke, or where stroke is a possible side effect of therapy. PMID- 12558470 TI - Cost-minimisation study of dorzolamide versus brinzolamide in the treatment of ocular hypertension and primary open-angle glaucoma: in four European countries. AB - OBJECTIVE: Cost is an issue when prescribing two drugs with equivalent efficacy. We compared the direct medical costs of topical brinzolamide 1% (twice a day or three times daily) with topical dorzolamide 2% (twice a day or three times daily) in France, Italy, Portugal and Spain in patients with ocular hypertension or primary open-angle glaucoma. DESIGN AND SETTING: Three double-blind, controlled, randomised trials (with a study duration of 3 months) compared the response rate of brinzolamide twice a day or three times daily versus dorzolamide three times daily, and the response rate of brinzolamide-timolol twice a day versus a dorzolamide-timolol combination twice a day. A fourth double-blind randomised trial (with a duration of 12 months) compared brinzolamide twice a day and three times daily with timolol monotherapy. Local tolerance was compared in two dedicated studies. Rates of switching to a new medication regimen were evaluated through a US health maintenance organisation database. In case of treatment failure, the patients were treated with latanoprost. A model was developed to value direct medical costs over 3 months. The economic perspective was that of the third-party payer and the patient, and included direct medical costs (reimbursed part plus co-payment). PATIENTS: Patients with ocular hypertension and/or primary open-angle glaucoma who had not responded to or could not tolerate beta-blocker therapy. OUTCOME MEASURE: The daily direct medical costs of therapy with the two drugs. RESULTS: As monotherapy, brinzolamide twice daily and three times daily was found to be as efficacious as dorzolamide three times a day. Brinzolamide twice daily plus timolol was also as efficacious as a combination of dorzolamide and timolol twice a day. Stinging of the eye upon instillation with brinzolamide was experienced by fewer patients than with dorzolamide (p < 0.0001). The likelihood of patients treated with dorzolamide changing therapy was 1.28 times greater than that for those treated with brinzolamide. The size of the brinzolamide drop is 18.7% smaller than that of dorzolamide allowing seven more therapy days per bottle with brinzolamide twice daily than with dorzolamide monotherapy, and five more days when brinzolamide is used three times a day. The direct medical costs for patients treated with brinzolamide were lower in all four European countries when drop size was taken into account than for those treated with dorzolamide. Sensitivity analyses confirmed the robustness of our findings. CONCLUSION: Because brinzolamide can be prescribed twice daily in monotherapy and because fewer patients treated with brinzolamide switch therapy due to local intolerance, our model suggests that brinzolamide is a cost-saving alternative to dorzolamide. PMID- 12558472 TI - Access block: problems and progress. PMID- 12558471 TI - Bedside rapid flu test and zanamivir prescription in healthy working adults: a cost-benefit analysis. AB - BACKGROUND: Zanamivir, a neuraminidase inhibitor, reduces the number of days of illness in influenza-positive patients. New bedside rapid flu tests (RFT) should increase the number of influenza-positive patients whom receive zanamivir appropriately. OBJECTIVE: To estimate the economic effects of implementing RFT and zanamivir among unvaccinated healthy working adults who consult within 2 days of the onset of influenza-like symptoms. METHODS: We constructed a decision tree to perform a cost-benefit analysis from a societal perspective. Clinical outcome, i.e. number of influenza days averted, and societal costs were compared for three strategies: RFT and conditional zanamivir prescription;systematic zanamivir prescription; and no zanamivir. A two-way sensitivity analysis was performed including the proportion of influenza-positive patients. RESULTS: During influenza epidemics, systematic zanamivir prescription provided the best health outcome (0.81 influenza days averted) and minimised societal costs (reduced by 29.80 US dollars per person compared with no zanamivir; 1999 values). RFT with conditional zanamivir averted 0.65 influenza days and saved 14.40 US dollars per person. When the proportion of influenza-positive patients was under 39%, the no zanamivir strategy yielded the greatest societal savings; otherwise, systematic zanamivir was the dominant strategy. Medical costs associated with no zanamivir were 88.70 US dollars per patient consulting with influenza-like illness, and increased to 125.50 US dollars with systematic zanamivir and to 127.60 US dollars with RFT and conditional zanamivir. CONCLUSIONS: Due to poor sensitivity of current RFT, systematic zanamivir prescription without RFT for unvaccinated healthy working adults should be recommended during influenza epidemics. PMID- 12558473 TI - Cardiovascular risk factors: when should we treat? PMID- 12558474 TI - Responses to access block in Australia: Australian Capital Territory. PMID- 12558476 TI - Responses to access block in Australia: Royal North Shore Hospital. PMID- 12558475 TI - Responses to access block in Australia: the Queen Elizabeth Hospital Medical Division. PMID- 12558477 TI - Responses to access block in Australia: Queensland. PMID- 12558478 TI - Responses to access block in Australia: Royal Perth Hospital. PMID- 12558479 TI - Responses to access block in Australia: Royal Melbourne Hospital. PMID- 12558480 TI - Responses to access block in Australia: The Alfred Hospital. PMID- 12558481 TI - Risk functions for prediction of cardiovascular disease in elderly Australians: the Dubbo Study. AB - OBJECTIVES: To evaluate a Framingham risk function for coronary heart disease in an elderly Australian cohort and to derive a risk function for cardiovascular disease (CVD) in elderly Australians. DESIGN AND SETTING: Analysis of data from a prospective cohort study (the Dubbo Study) in a semi-urban town (population, 34 000). PARTICIPANTS: 2805 men and women 60 years and older living in the community, first assessed in 1988, and a subcohort of 2102 free of CVD at study entry. MAIN OUTCOME MEASURES: Incidence of CVD (myocardial infarction, coronary death or stroke) over 5 and 10 years. RESULTS: A Framingham risk function assessing "hard" coronary heart disease (ie, myocardial infarction or coronary death) accurately predicted 10-year incidence in men and women aged 60-79 years who were free of prevalent CVD or diabetes at study entry. In a multiple logistic model, CVD incidence was significantly predicted by age, sex, taking antihypertensive medication, blood pressure, smoking, total cholesterol level and diabetes. For a given age and cholesterol level, CVD risk over 5 years was doubled in the presence of antihypertensive medication or diabetes, increased by 50% with cigarette smoking, and halved in women compared with men. CONCLUSIONS: We have derived a simple CVD risk function specifically for elderly Australians that employs risk factors readily accessible to all medical practitioners. PMID- 12558482 TI - Appropriateness of platelet, fresh frozen plasma and cryoprecipitate transfusion in New South Wales public hospitals. AB - OBJECTIVES: To estimate the appropriateness of transfusions of platelets, fresh frozen plasma (FFP) and cryoprecipitate using National Health and Medical Research Council and Australasian Society for Blood Transfusion guidelines (NHMRC/ASBT 2002). DESIGN AND SETTING: Three separate retrospective surveys of medical records from 1 January to 31 August 2000 (1147 transfused patients) from 14 hospitals selected randomly from all public hospitals that use these blood products in New South Wales: five tertiary referral, five major metropolitan, and four major rural (base) hospitals. MAIN OUTCOME MEASURES: Proportion of potentially inappropriate transfusions. RESULTS: 33% (136/414) of platelet, 37% (248/669) of FFP and 62% (37/60) of cryoprecipitate transfusions were assessed as inappropriate. By hospital type, 29% (75/259) of platelet transfusions were inappropriate at tertiary referral hospitals, 51% (40/78) at major urban hospitals, and 27% (21/79) at major rural hospitals. For FFP, 36% (112/313), 37% (80/216) and 39% (55/140) were inappropriate for referral, urban and rural hospitals, respectively. Cryoprecipitate was used almost exclusively at tertiary referral hospitals. CONCLUSIONS: In terms of the NHMRC/ASBT guidelines on use of blood products, there is considerable inappropriate transfusion of platelets, FFP and cryoprecipitate in NSW public hospitals. PMID- 12558483 TI - The decline in bulk-billing and increase in out-of-pocket costs for general practice consultations in rural areas of Australia, 1995-2001. AB - OBJECTIVE: To describe the changes in bulk-billing and out-of-pocket costs for Australian general practice consultations over the period 1995-2001. DESIGN: Retrospective analysis of 1996-2001 survey data from the Australian Longitudinal Study on Women's Health (ALSWH), linked with Medicare and Department of Veterans' Affairs (DVA) data on general practice consultations from 1995 to 2001. PARTICIPANTS: 22 633 women who gave consent to linkage of their ALSWH data with Medicare/DVA records. In 1996, women in the "young" cohort (n = 6219) were aged 18-23 years, those in the "mid-age" cohort (n = 8883) were aged 45-50 years, and those in the "older" cohort (n = 7531) were aged 70-75 years. OUTCOME MEASURES: Out-of-pocket costs paid by patients for general practice consultations, by calendar year, urban/rural area of residence, age, frequency of attendance, self rated health, and education level. RESULTS: For each age group and year studied, the use of bulk-billing was lower in rural areas than in urban areas. For example, in 2000, the percentage of women in rural and urban areas, respectively, who had all their general practice consultations bulk-billed was 31% v 52% (young women), 24% v 45% (mid-age women) and 58% v 79% (older women). There has been a steady decline in bulk-billing for general practice consultations in rural areas since 1995. The average out-of-pocket cost per consultation for women in rural areas was higher than the cost for women living in urban areas. After adjusting for age, health and socioeconomic factors, women living in urban areas were more than twice as likely to have all their consultations bulk-billed as women living in rural areas: odds ratio (OR), 2.4 (95% CI, 2.1-2.7) (young women); OR, 2.5 (95% CI, 2.3-2.8) (mid-age women); OR, 2.6 (95% CI, 2.3-2.9) (older women). CONCLUSIONS: In Australia, the geographic differential in the cost of general practice consultations is widening. Policy changes are required to enable women in rural and remote areas to have access to affordable healthcare services. PMID- 12558484 TI - Embryo donation at an Australian university in-vitro fertilisation clinic: issues and outcomes. AB - OBJECTIVES: To review the choices of couples relinquishing frozen embryos and the outcomes of embryo donation at a major in-vitro fertilisation (IVF) clinic. DESIGN AND SETTING: Retrospective audit of 11.5 years of data (1991-2002) from the Monash University IVF clinic, Melbourne. PARTICIPANTS: Couples who make decisions regarding the fate of their frozen embryos, and recipient couples taking part in embryo adoption. MAIN OUTCOME MEASURES: Couples' choices with regard to the fate of their frozen embryos, and the outcome of donated embryo treatment cycles. RESULTS: Of 1246 couples relinquishing frozen embryos, 1116 (89.5%) opted to discard rather than donate their embryos. Sixty-six per cent of donated embryos survived thawing. From donated-embryo transfer to 50 women in 92 cycles, a 17.4% pregnancy rate per transfer cycle was achieved, and 10 women delivered 11 healthy babies at term. At the time of our audit there were 98 couples on the waiting list to adopt embryos. CONCLUSIONS: It is worth considering how couples can be encouraged to donate rather than discard their surplus frozen embryos. An educational program on relevant legal, social and clinical issues may facilitate this. PMID- 12558485 TI - "Munchausen by proxy syndrome": not only pathological parenting but also problematic doctoring? AB - Certain social expectations of medicine combine with characteristics of subspecialised technological paediatrics to facilitate the form of child abuse labelled "Munchausen by proxy syndrome". Examining this form of child abuse highlights possible shortcomings of medical practice. The primary medical tasks of diagnosing and curing illness and of preventing suffering are sometimes overridden by other motivations of which doctors may not be fully aware. More open discussion of what motivates health professionals in their work may improve medical practice and lead to a reduced incidence of Munchausen by proxy syndrome. PMID- 12558486 TI - "Munchausen by proxy syndrome": not just pathological parenting but also problematic doctoring? Another view. PMID- 12558487 TI - Early clinical clues to meningococcaemia. AB - Meningococcal septicaemia has high mortality, especially when the diagnosis is delayed or missed. Early recognition is not always straightforward, as classic clinical features may be absent or overlooked at initial presentation. Septicaemia without focal infection accounts for 15%-20% of cases of meningococcal disease and is the most worrisome manifestation in terms of diagnosis and outcome; in contrast, meningococcal meningitis is usually straightforward to diagnose, with a relatively good prognosis. Useful early clinical clues to meningococcaemia include: - a haemorrhagic (petechial or purpuric) rash; - blanching macular or maculopapular rash that appears in first 24 hours of illness; - true rigors; - severe pain in extremities, neck or back; vomiting, especially in association with headache or abdominal pain; rapid evolution of the illness; - concern of parents, relatives or friends; - patient age (highest incidence at age 3-12 months, followed by 1-4 and then 15-19 years); and - contact with a patient with meningococcal disease. In addition to specific clues, clinicians should look at the whole pattern of the illness. Timely clinical review is essential if there is doubt about the diagnosis. In any acutely febrile patient, it is prudent to ask "Why is this patient seeking help now?", then "Could this patient have meningococcaemia?". PMID- 12558488 TI - Fatal envenomation by jellyfish causing Irukandji syndrome. PMID- 12558489 TI - Chemical-biological-radiological (CBR) response: a template for hospital emergency departments. PMID- 12558490 TI - Trusting numbers: uncertainty and the pathology laboratory. PMID- 12558491 TI - eTG complete. PMID- 12558493 TI - Ross River virus--are we wasting money doing tests? PMID- 12558494 TI - Itching bites may limit Ross River virus infection. PMID- 12558495 TI - Improving doctors' letters. PMID- 12558496 TI - Correlates of endothelial function and their relationship with inflammation in patients with familial hypercholesterolaemia. AB - Atherosclerosis is characterized by a low-grade systemic inflammatory response and endothelial dysfunction. The aim of the present study was to investigate a possible relationship between systemic markers of inflammation, serum markers of endothelial activation and endothelium-dependent vasodilatation in a group of high-risk patients, and to evaluate the effects of intervention with high doses of simvastatin on these parameters. In patients with heterozygous familial hypercholesterolaemia, without atherosclerotic events, flow-mediated vasodilatation (FMD) of the brachial artery was measured after a wash-out period for lipid-lowering drugs (baseline) and after 6 weeks of treatment with simvastatin 80 mg daily. Levels of C-reactive protein (CRP), soluble intercellular cell-adhesion molecule (s-ICAM) and soluble E-selectin (s-E selectin) were determined at baseline and again after 6 weeks and 12 months of therapy. A total of 35 subjects participated in the study (mean age 42 years; 60% male). When divided into tertiles according to FMD (<3.9%, 3.9-9.0% and >9.0%), no differences in levels of CRP, s-ICAM-1 and/or s-E-selectin were detected between the groups. Moreover, no changes in FMD, levels of CRP or levels of s ICAM-1 and/or s-E-selectin were found during treatment with simvastatin. We conclude that endothelial function, as reflected by FMD, does not seem to be related to markers of inflammation in familial hypercholesterolaemia subjects at high risk of, but without clinically overt signs of, atherosclerosis. Moreover, aggressive lipid-lowering therapy with simvastatin does not result in improved endothelial function or in a reduction of markers of inflammation in these patients. PMID- 12558498 TI - Structural requirements of human DNase II alpha for formation of the active enzyme: the role of the signal peptide, N-glycosylation, and disulphide bridging. AB - DNase II alpha (EC 3.1.22.1) is an endonuclease, which is active at low pH, that cleaves double-stranded DNA to short 3'-phosphoryl oligonucleotides. Although its biochemistry is well understood, its structure-activity relationship has been largely unexamined. Recently, we demonstrated that active DNase II alpha consists of one contiguous polypeptide, heavily glycosylated, and containing at least one intrachain disulphide linkage [MacLea, Krieser and Eastman (2002) Biochem. Biophys. Res. Commun. 292, 415-421]. The present paper describes further work to examine the elements of DNase II alpha protein required for activity. Truncated forms and site-specific mutants were expressed in DNase II alpha-null mouse cells. Results indicate that the signal-peptide leader sequence is required for correct glycosylation and that N-glycosylation is important for formation of the active enzyme. Despite this, enzymic deglycosylation of wild-type protein with peptide N-glycosidase F reveals that glycosylation is not intrinsically required for DNase activity. DNase II alpha contains six evolutionarily conserved cysteine residues, and mutations in any one of these cysteines completely ablated enzymic activity, consistent with the importance of disulphide bridging in maintaining correct protein structure. We also demonstrate that a mutant form of DNase II alpha that lacks the purported active-site His(295) can still bind DNA, indicating that this histidine residue is not simply involved in DNA binding, but may have a direct role in catalysis. These results provide a more complete model of the DNase II alpha protein structure, which is important for three-dimensional structural analysis and for production of DNase II alpha as a potential protein therapeutic for cystic fibrosis or other disorders. PMID- 12558499 TI - Cleavage of fragments containing DNA mismatches by enzymic and chemical probes. AB - We prepared synthetic 50-mer DNA duplexes, each containing four mismatched base pairs in similar positions. We examined their cleavage by DNases I and II, micrococcal nuclease (MNase), methidiumpropyl-EDTA-Fe(II) [MPE-Fe(II)] and hydroxyl radicals. We find that single mismatches only produce subtle changes in the DNase I-cleavage pattern, the most common of which is attenuated cleavage at locations 2-3 bases on the 3'-side of the mismatch. Subtle changes are also observed in most of the DNase II-cleavage patterns, although GT and GG inhibit the cleavage over longer regions and generate patterns that resemble footprints. MNase cleaves the heteroduplexes at the mismatches themselves (except for CC), and in some cases cleaves CpG and CpC steps. None of the mismatches causes any change in the cleavage patterns produced by hydroxyl radicals or MPE-Fe(II). We also examined the cleavage patterns of fragments containing tandem GA mismatches in the sequences RGAY/RGAY and YGAR/YGAR (R, purine; Y, pyrimidine). RGAY causes only subtle changes in the cleavage patterns, which are similar to those seen with single mismatches, except that there are no changes in MNase cleavage. However, YGAR inhibits DNases I and II cleavage over 4-6 bases, and attenuates MPE-Fe(II) and hydroxyl radical cleavage at 2 bases. These changes suggest that this mismatch has a more pronounced effect on the local DNA structure. These changes are discussed in terms of the structural and dynamic effects of each mismatch. PMID- 12558497 TI - Killing tumours by ceramide-induced apoptosis: a critique of available drugs. AB - Over 1000 research papers have described the production of programmed cell death (apoptosis) by interventions that elevate the cell content of ceramide (Cer). Other interventions, which lower cellular Cer, have been found to interfere with apoptosis induced by other agents. Some studies have shown that slowing the formation of proliferation-stimulating sphingolipids also induces apoptosis. These relationships are due to the two different aspects of Cer: Cer itself produces apoptosis, but metabolic conversion of Cer into either sphingosine 1 phosphate or glucosphingolipids leads to cell proliferation. The balance between these two aspects is missing in cancer cells, and yet intervention by stimulating or blocking only one or two of the pathways in Cer metabolism is very likely to fail. This results from two properties of cancer cells: their high mutation rate and the preferential survival of the most malignant cells. Tumours treated with only one or two drugs that elevate Cer can adjust the uncontrolled processes to either maintain or to 'aggravate' the excessive growth, angiogenesis and metastasis characteristics of tumours. These treatments might simply elevate the production of growth factors, receptors and other substances that reduce the effectiveness of Cer. Tumour cells that do not adapt in this way undergo apoptosis, leaving the adapted cells free to grow and, ultimately, to 'subdue' their host. Thus it is important to kill every type of cancer cell present in the tumour rapidly and simultaneously, using as many different agents to control as many pathways as possible. To aid this approach, this article catalogues many of the drugs that act on different aspects of Cer metabolism. The techniques described here may lead to the development of practical chemotherapy for cancer and other diseases of excess proliferation. PMID- 12558500 TI - Troponin C in different insect muscle types: identification of two isoforms in Lethocerus, Drosophila and Anopheles that are specific to asynchronous flight muscle in the adult insect. AB - The indirect flight muscles (IFMs) of Lethocerus (giant water bug) and Drosophila (fruitfly) are asynchronous: oscillatory contractions are produced by periodic stretches in the presence of a Ca(2+) concentration that does not fully activate the muscle. The troponin complex on thin filaments regulates contraction in striated muscle. The complex in IFM has subunits that are specific to this muscle type, and stretch activation may act through troponin. Lethocerus and Drosophila have an unusual isoform of the Ca(2+)-binding subunit of troponin, troponin C (TnC), with a single Ca(2+)-binding site near the C-terminus (domain IV); this isoform is only in IFMs, together with a minor isoform with an additional Ca(2+) binding site in the N-terminal region (domain II). Lethocerus has another TnC isoform in leg muscle which also has two Ca(2+)-binding sites. Ca(2+) binds more strongly to domain IV than to domain II in two-site isoforms. There are four isoforms in Drosophila and Anopheles (malarial mosquito), three of which are also in adult Lethocerus. A larval isoform has not been identified in Lethocerus. Different TnC isoforms are expressed in the embryonic, larval, pupal and adult stages of Drosophila; the expression of the two IFM isoforms is increased in the pupal stage. Immunoelectron microscopy shows the distribution of the major IFM isoform with one Ca(2+)-binding site is uniform along Lethocerus thin filaments. We suggest that initial activation of IFM is by Ca(2+) binding to troponin with the two-site TnC, and full activation is through the action of stretch on the complex with the one-site isoform. PMID- 12558501 TI - Characterization of poultry egg-white avidins and their potential as a tool in pretargeting cancer treatment. AB - Chicken avidin and bacterial streptavidin are proteins used in a wide variety of applications in the life sciences due to their strong affinity for biotin. A new and promising use for them is in medical pretargeting cancer treatments. However, their pharmacokinetics and immunological properties are not always optimal, thereby limiting their use in these applications. To search for potentially beneficial new candidates, we screened egg white from four different poultry species for avidin. Avidin proteins, isolated from the duck, goose, ostrich and turkey, showed a similar tetrameric structure, similar glycosylation and stability against both temperature and proteolytic activity of proteinase K as chicken avidin. Biotin-binding properties of these avidins, measured using IAsys optical biosensor, were similar to those found in avidin from the chicken. Three of these novel avidins, however, showed different immunological cross reactivities when compared with chicken avidin. The patient sera responses to duck, goose and ostrich avidins were also lower than those observed for chicken and turkey avidins. Our findings suggest that the use of these proteins offers advantages over chicken avidin and bacterial streptavidin in pretargeting applications. PMID- 12558502 TI - Protein kinase CK2 phosphorylates Hsp105 alpha at Ser509 and modulates its function. AB - The 105 kDa heat-shock protein (Hsp) Hsp105 alpha is a mammalian stress protein that belongs to the HSP105/HSP110 family. We have shown previously that Hsp105 alpha exists as non-phosphorylated and phosphorylated forms in vivo, and is phosphorylated by protein kinase CK2 (CK2) in vitro. In this study, to elucidate the role of phosphorylation of Hsp105 alpha, we first analysed the site of phosphorylation of Hsp105 alpha by CK2. Peptide mapping analysis of Hsp105 alpha phosphorylated by CK2 and in vitro phosphorylation experiments using various deletion and substitution mutants of Hsp105 alpha revealed that Hsp105 alpha is phosphorylated at Ser(509) in the beta-sheet domain. Furthermore, Ser(509) in Hsp105 alpha was also phosphorylated in mammalian COS-7 cells, although other sites were phosphorylated as well. Next, we examined the effects of phosphorylation of Hsp105 alpha on its functions using CK2-phosphorylated Hsp105 alpha. Interestingly, Hsp105 alpha suppressed 70 kDa heat-shock cognate protein (Hsc70)-mediated protein folding, whereas the phosphorylation of Hsp105 alpha at Ser(509) abolished the inhibitory activity of Hsp105 alpha in vitro. In accordance with these findings, wild-type Hsp105 alpha, which was thought to be phosphorylated in vivo, had no effect on Hsp70-mediated refolding of heat denatured luciferase, whereas a non-phosphorylatable mutant of Hsp105 alpha suppressed the Hsp70-mediated refolding of heat-denatured luciferase in mammalian cells. Thus it was suggested that CK2 phosphorylates Hsp105 alpha at Ser(509) and modulates the function of Hsp105 alpha. The regulation of Hsp105 alpha function by phosphorylation may play an important role in a variety of cellular events. PMID- 12558503 TI - U2 small nuclear ribonucleoprotein particle (snRNP) auxiliary factor of 65 kDa, U2AF65, can promote U1 snRNP recruitment to 5' splice sites. AB - The splicing factor U2AF(65), U2 small nuclear ribonucleoprotein particle (snRNP) auxillary factor of 65 kDa, binds to pyrimidine-rich sequences at 3' splice sites to recruit U2 snRNP to pre-mRNAs. We report that U2AF(65) can also promote the recruitment of U1 snRNP to weak 5' splice sites that are followed by uridine-rich sequences. The arginine- and serine-rich domain of U2AF(65) is critical for U1 recruitment, and we discuss the role of its RNA-RNA annealing activity in this novel function of U2AF(65). PMID- 12558504 TI - Continuous production of a peptidic fraction containing the intermediate opioid peptide LVV-haemorphin-7 (LVVh-7) by peptic hydrolysis of bovine haemoglobin in a continuous membrane reactor. AB - Peptic hydrolysis of native bovine haemoglobin at pH 3 yields the LVV-haemorphin 7 (Leu-Val-Val-Tyr-Pro-Trp-Thr-Gln-Arg-Phe; LVVh-7) opioid peptide corresponding to the residues-31-40 fragment of the beta-chain of haemoglobin. This peptide is intermediate in the course of batch hydrolysis and is rapidly degraded. Indeed, it shows an optimum at 3% degree of hydrolysis (i.e. 2 min of reaction time). The hydrolysis was carried out in a continuous membrane reactor with a space time (ratio of the flux to the reactor volume) set to 2 min (corresponding to optimum LVVh-7 production). This process allows the continuous production of a constant fraction of intermediate peptides containing LVVh-7 for 48 min. PMID- 12558524 TI - B2 bradykinin receptor mediates the stimulatory effect of bradykinin on rat germ cell proliferation in vitro. AB - The contribution of B1 and B2 bradykinin receptors to germ cell proliferation was studied by using in vitro organ cultures of testicular fragments of 3.5-day-old rats in the presence of 3H thymidine. Different combinations of agonists and antagonists of B1 and B2 receptors exerted differential mitogenic effect on pro spermatogonial cells. Application of bradykinin (B2 receptor agonist) alone induced a threefold increase in germ cell labelling index compared with the control, whereas des-Arg9-bradykinin (B1 receptor agonist) caused weak stimulation (24%) on spermatogonial mitotic activity. The bradykinin-induced germ cell proliferation was significantly affected by B2 receptor antagonist (HOE-140) but not by B1 receptor antagonist. When B1 or B2 receptor antagonists were applied with des-Arg9-bradykinin, the germ cell labelling indices were nonsignificantly different compared with those of B1 receptor agonist only. The present findings suggest that B2 receptor is involved in mediating the stimulatory effect of bradykinin on germ cell proliferation and therefore bradykinin might be an important local testicular factor in the regulation of spermatogonial division and germ cell number. PMID- 12558525 TI - Structure and signalling pathways of kinin receptors. AB - Kinins are peptide hormones that transmit their biological effects via G protein coupled receptors. They are generated by kallikrein-mediated proteolysis of their precursors, the kininogens. Kinins have been implicated in the regulation of blood pressure, pain sensation and cell growth. Interestingly, all components of the kallikrein-kinin system have also been localized in testis. Effects of kallikrein and bradykinin on pre-spermatogonial cell proliferation and on sperm motility suggest a regulatory function of kinins and their cognate receptors in the male reproductive system. This review is dedicated to summarize the current knowledge about structure, signal transduction and regulation of kinin receptors. Particular emphasis will be given to the kinin-induced activation of the mitogen activated protein kinase cascade which might represent an important signalling pathway involved in regulation of spermatogenesis and sperm function. PMID- 12558526 TI - Tissue kallikrein and bradykinin B2 receptors in the reproductive tract of the male rat. AB - The setting of a local tissue kallikrein kinin system (tKKS) within the reproductive organs of the male rat was investigated by analysing bradykinin subtype 2 receptor (B2R) gene expression and cellular distribution of B2R protein and the kinin-liberating protease tissue kallikrein (tK). Reverse transcription polymerase chain reaction showed B2R expression in testis, epididymis and prostate from prepubertal and sexually mature rats. In mature testis, in situ hybridization and immunohistochemistry localized B2R mRNA and protein besides endothelial cells of blood vessels exclusively on pachytene spermatocytes and round and elongated spermatids. B2R expression within the seminiferous tubules was found to be dependent on the stage of the spermatogenic cycle. In pre pubertal rat testis, B2R mRNA and protein were additionally located in peritubular cells. In the testis, specific staining for tK occurred in addition to endothelial cells of blood vessels on the acrosomal cap of round and elongated spermatids. This immunostaining was also stage-dependent. In the epididymis, tK was detected on epithelial cells near the apical surface. The stage-dependent specific expression of tK and bradykinin B2Rs in developing germ cells and peritubular cells suggests a potential role of the tKKS in the local regulation of spermatogenesis and seminiferous tubule function. PMID- 12558527 TI - Isoforms of angiotensin I-converting enzyme in the development and differentiation of human testis and epididymis. AB - Angiotensin I-converting enzyme (ACE; CD143, Kininase II, EC 3.4.15.1) is known to be crucial for male fertility in animal models. We therefore studied its testicular (tACE) and somatic (sACE) isoforms in foetal and adult human testis and epididymis using monoclonal antibodies and cRNA probes. During spermatogenesis, tACE was found only in differentiating germ cells and was the only isoform within the seminiferous tubules of adult men. Although tACE mRNA was present in spermatocytes, tACE protein was initially found in post-meiotic step 3 spermatids and increased markedly during further differentiation. The enzyme was strictly confined to the adluminal membrane site of elongating spermatids and was localized at the neck and midpiece region of released and ejaculated spermatozoa. In contrast, sACE was expressed heterogeneously in Leydig cells and endothelial cells of the testicular interstitium, and homogeneously along the luminal surface of epithelial cells lining the ductuli efferents, corpus and cauda of epididymis, and vas deferens. The cell- and site-restricted pattern of sACE corresponded to that found in foetal tissues except an additional and transient expression of sACE in foetal germ cells and foetal Sertoli cells. Our study documents for the first time in humans the regulation and unique cellular distribution of ACE isoforms during the ontogenesis of the lower male genital tract. PMID- 12558528 TI - Effects of kinins on mammalian spermatozoa and the impact of peptidolytic enzymes. AB - Effects of kinins, mainly bradykinin (Bk), and other components of the kallikrein kinin system on sperm motility and further fertility-related functions have been described repeatedly. However, reported data are in part controversial and the mechanism of kinin effects on sperm motility is not yet understood. In the present report we describe a significant promoting effect of Bk on sperm motility at subnanomolar concentrations. This effect was stabilized and even increased by suppression of Bk hydrolysis in semen samples. As sperm membrane-bound angiotensin-converting enzyme and neutral metalloendopeptidase are mainly involved in Bk hydrolysis, an effective cocktail of enzyme inhibitors promoting the sperm motility consists of phosphoramidon and lisinopril (both at 10-7 m). The effects of Bk on sperm cells are not mediated by the B2 Bk receptor. Using several biochemical, molecular and genetic methods we could not detect any Bk receptor on spermatozoa. PMID- 12558529 TI - Immune-endocrine interactions and Leydig cell function: the role of cytokines. AB - Inflammatory disease is known to affect male reproductive function and fertility. Male accessory gland infections (MAGI) account for almost 15% of all cases of male infertility seen in infertility clinics. Infections of the male accessory glands are associated with increased counts of white blood cells in semen and elevated levels of pro-inflammatory cytokines in the semen and the testis. Numerous studies have underscored the importance of cytokines in the regulation of testicular and glandular function during pathophysiological events as well as under normal physiological conditions when cytokines act as growth and differentiation factors. The purpose of this paper is to particularly review the role of cytokines in the regulation of Leydig cell function in the testis primarily under pathophysiological conditions, and also considers clinical investigations that help to improve the evaluation and treatment of male infertility. PMID- 12558530 TI - The Fas system in the seminiferous epithelium and its possible extra-testicular role. AB - The Fas system is involved in the control of immune system homeostasis and nonfunctional Fas system leads to autoimmune disease in mice and humans. The Fas system is a mechanism through which cells expressing Fas ligand (FasL) induce apoptosis of Fas expressing cells. In mouse and rat, the testis represents the main source of constitutive FasL in the body. The roles so far proposed for this molecule in the testis, such as maintenance of immunoprivilege and regulation of physiological germ cell apoptosis, need to be reconsidered as both hypotheses are based on an erroneous cellular location of FasL in the seminiferous epithelium. Recently, we demonstrated that in rodents FasL mRNA is present in germ cells and not in Sertoli cells, and that FasL protein is displayed on the surface of spermatozoa. Here we propose that, for the mouse spermatozoa, the FasL may represent a self-defence mechanism against lymphocytes present in the female genital tract. To verify this hypothesis, we performed crossings between males gld, with nonfunctional FasL, and syngenic or nonsyngenic females. We observed a significant decrease of litter size in outbred crossings with gld males compared with wild-type males, suggesting a possible role of FasL in immunoprotection of the sperm in the female genital tract. The possibility that in humans, by analogy with mouse, FasL plays a self-protective role for the spermatozoon cannot be excluded, and awaits experimental information on the expression of FasL on human sperm cells. PMID- 12558531 TI - Molecular mechanisms utilized by alternative c-kit gene products in the control of spermatogonial proliferation and sperm-mediated egg activation. AB - The c-kit proto-oncogene plays a dual role in the control of male fertility in mice through two alternative gene products: (1). c-kit [the transmembrane tyrosine kinase receptor for stem cell factor (SCF)], which is expressed and functional in differentiating spermatogonia of the postnatal testis, in which c kit is essential for pre-meiotic proliferation; and (2). tr-kit, an intracellular protein which is specifically accumulated during spermiogenesis through the use of an alternative intronic promoter, and which is able to trigger mouse egg activation when microinjected into the cytoplasm of metaphase II arrested oocytes. Here, we summarize the most recent findings about the molecular pathways through which c-kit regulates cell cycle progression in mitotic germ cells, and those through which sperm-derived tr-kit triggers parthenogenetic completion of meiosis II and pronuclear formation in microinjected mouse eggs. PMID- 12558532 TI - Development of the foetal and neonatal testis. AB - The foetal testis originates from a proliferation of the mesonephric and the coelomic epithelia which are colonized by the primordial germ cells. In the foetal testis, the development and functions of the three main cell type precursors (Leydig, Sertoli and germ cells) do not depend upon gonadotropins. Numerous intra- and extra-testicular factors are candidates for the control of its development and functions. To study the potential involvement of these factors, we developed an organotypic culture system. In absence of any growth factors or hormone, this system allows a development of the three main cell types which mimics that observed in vivo. The effects of different regulators (gonadotropin-releasing hormone, follicle-stimulating hormone, transforming growth factor-beta, insulin-like growth factor-I, anti-Mullerian hormone, retinoic acid, oestrogens) were tested in this system. Whether or not some of the effects observed in vitro have a physiological relevance was evaluated using appropriate transgenic mice. It is concluded that the foetal testis cannot be considered as an adult mini-testis since it has a specific physiology which largely differs from that of the immature or adult testis. PMID- 12558533 TI - New trends in molecular andrology. Proceedings of a workshop. Giessen, Germany. May 3-4, 2002. PMID- 12558536 TI - Antenatal screening for postnatal depression: a systematic review. AB - OBJECTIVE: To describe the screening properties of the antenatal tools which have been developed to predict depression after birth and to summarize the implications of the findings for antenatal screening. METHOD: Systematic review and secondary analysis of published papers. RESULTS: Sixteen studies including sufficient data for the calculation of screening properties were identified. The majority developed a study-specific screening instrument. Outcome assessments used the Edinburgh Postnatal Depression Scale or standardized diagnostic psychiatric interviews, or both. In the two largest population-based studies, the proportion of women screening as positive ('at risk' of postnatal depression) was 16 and 52%, respectively, and of these only 35 and 8% actually developed depression after birth. CONCLUSION: No screening instrument(s) met the criteria for routine application in the antenatal period. Factors that may have affected poor sensitivity and positive predictive values include the exclusion of key domains in predicting risk, particularly personality, a past history of abuse and postnatal events, the contribution of which may be being under-estimated in studies evaluating antenatal risk prediction tools. PMID- 12558535 TI - Methodological pitfalls in early detection studies - the NAPE Lecture 2002. Nordic Association for Psychiatric Epidemiology. AB - OBJECTIVE: To identify and discuss methodological pitfalls that may help explain why many questions around early detection (ED) and duration of untreated psychosis (DUP) are still unsolved. METHOD: This paper concentrates on pitfalls in the following areas: sampling, measurement and data analyses. RESULTS: The main problems seem to be: SAMPLING: Referral bias, exclusion of patients, patient refusal, and patients lost to follow-up. MEASUREMENT: Reliability, which is particularly cogent for multisite investigations, and validity, which includes: Start of illness, start of psychosis, diagnoses, start of treatment, the relationship between ED and DUP and choice of outcome measures. Data Analyses: Overlooking threshold effects of DUP, improper control for baseline scores, and lack of control for confounders. CONCLUSION: Methodological pitfalls may bias ED studies. Several pitfalls are unavoidable, but proper design and quality assurance can reduce their impact. Researchers ought to identify the pitfalls, and to estimate and discuss their influence. PMID- 12558537 TI - Protracted acute stress reaction following an earthquake. AB - OBJECTIVE: To verify the official criteria of ICD-10 diagnosis of acute stress reaction (ASR) among earthquake victims. METHOD: Data on psychosocial variables and the diagnosis of ASR were collected from 91 subjects. RESULTS: The diagnosis of ASR was made in 70% of the sample. However, the restrictive duration criterion (i.e. symptoms not exceeding the first 48 h) was fulfilled only in 10%; in the remaining 60% the symptoms lasted for about another week. Thus, 60% of the sample constituted a group with protracted ASR (PASR) and 40% a group without it (n PASR). Medical history was more frequent among PASR than n-PASR; PASR showed higher anxiety levels (both pre- and post-disaster) than n-PASR; finally, the persistence of ASR related positively to the fear of death at the time of the earthquake and pre-disaster anxiety levels. CONCLUSION: As in the vast majority of earthquake victims the ASR is protracted beyond 48 h, a revision of ICD-10 guidelines should be considered. PMID- 12558538 TI - Life events and depressiveness - the effect of adjustment for psychosocial factors, somatic health and genetic liability. AB - OBJECTIVE: To study the nature of the relationship of life events with depressiveness. METHOD: In 1990, 9947 Finnish adult twins aged 33-60 years participated in a questionnaire study, with no missing data on the analysed variables. The relationship between stressful life-events and depressiveness, based on Beck Depression Inventory (BDI) classified as normal, mild or moderate, was analysed using multinomial regression for all subjects with adjustment for the effect of age, sex, health status, social support, marital status, social class, and personality variables. These same factors were analyzed among 643 twin pairs discordant for depressiveness. RESULTS: High BDI score categories were strongly associated with stressful life events in all individuals and equally strongly within discordant twin pairs, both monozygotic and dizygotic. Poor somatic health and lack of social support also increased depressiveness. CONCLUSION: The effect of life events predicting depressiveness was independent of social support, somatic disease, sex and genetic liability. PMID- 12558539 TI - Functional and dysfunctional impulsivity: contribution to the construct validity. AB - OBJECTIVE: Impulsivity has been found to be an important trait of personality, whose consequences are not always negative although available questionnaires focused on its 'dysfunctional' aspect. METHOD: Dickman's Functional and Dysfunctional Impulsivity questionnaire has been translated into French, and filled out by students. The tetrachoric correlation matrices were factor analysed. RESULTS: The psychometric properties are very close to those of the English version, and we recovered both factors in males and females. The factor similarities between the genders (in our sample) and between the languages (English vs. French) are very good. The Dysfunctional scale is correlated with the Motor Impulsivity subscale of the Barratt Impulsiveness Scale, and both scales are grossly independent from Spielberger's Trait-Anxiety Inventory. CONCLUSION: Our results support a two-factor solution similar to the English one. Nevertheless, the validity of the functional factor remains to be investigated in further studies. PMID- 12558540 TI - Characteristics of HIV patients who attempt suicide. AB - OBJECTIVE: To examine the risk factors for suicidal behaviour in human immunodeficiency virus (HIV) positive patients. METHOD: HIV substance dependent positive patients who had attempted suicide were compared with HIV substance dependent positive patients who had never attempted suicide for suicide risk factors. RESULTS: Among the 149 HIV positive patients examined almost half had attempted suicide. Significantly more HIV positive patients who had attempted suicide were female. Attempters were significantly younger than non-attempters. Significantly more of the attempters had a family history of suicidal behaviour. Attempters also reported significantly more childhood trauma, scored significantly higher for neuroticism, had experienced significantly more comorbidity with depression, and more of them had received antidepressant medication. CONCLUSION: These data suggest that both distal and proximal risk factors are involved in suicidal behaviour in HIV positive substance dependent patients. PMID- 12558541 TI - Risk factors in neuroleptic malignant syndrome. A case-control study. AB - OBJECTIVE: To determine whether environmental temperature, agitation, neuroleptic use, mental retardation, and affective disorders were risk factors for neuroleptic malignant syndrome (NMS). METHOD: Cases and age- and sex-matched psychiatric controls admitted to a regional acute psychiatric unit over a 10-year period. RESULTS: Both uni- and multivariate analysis revealed statistically significant differences between patients with NMS (n=15) and controls (n=45) with regard to the presence of mental retardation, psychomotor agitation, and a number of variables relating to neuroleptic use (newly introduced or increased, intramuscular administration, and dosage). We found no differences between NMS patients and psychiatric controls in respect of changes in environmental temperature. CONCLUSION: Our study supports the need for caution when using intramuscularly administered, abruptly increasing, high-dose neuroleptics, particularly in mentally retarded or agitated patients, regardless of environmental temperature. PMID- 12558542 TI - Does extra staff change clinical practice? A prospective study of the impact of extra resources in mental health teams. AB - OBJECTIVE: To compare patterns of clinical activity amongst existing staff in two inner-city community mental health teams before and after their enhancement with extra resources using a new activity schedule containing 11 comprehensive and mutually exclusive practice categories. METHOD: Patterns of clinical activity amongst existing members of two inner-London community mental health teams were compared before and after the addition of extra resources. RESULTS: The amount of time spent in face-to-face contact with patients and carers showed no meaningful change following team enhancement. Patterns of clinical activity amongst team members remained relatively static. CONCLUSION: Adding resources to community mental teams without considering how to target the time released amongst existing staff may reduce their capacity to work more innovatively. PMID- 12558543 TI - Cerebral tissue alterations and daily life stress experience in psychosis. AB - OBJECTIVE: To examine whether the total volumes of cerebrospinal fluid (CSF), cerebral grey matter and white matter were correlated with the experience of environmental stress in daily life situations. METHOD: Twenty-seven patients with psychosis underwent magnetic resonance imaging scanning and a random time sampling self-assessment technique (Experience Sampling Method) to determine subjective daily life stress experiences. Total cerebral tissue volumes were derived from an automated segmentation procedure. RESULTS: CSF volume was positively associated with daily life event-related stress (beta=0.016, P=0.002), while the association with total white matter was negative (beta=-0.013, P=0.005). The effects were independent of each other and of total cerebral volume and other confounders. No large or significant association was found with grey matter volume. CONCLUSION: Subjective stress experience in daily life is associated with increased CSF and reduced white matter volumes in patients with psychosis, suggesting functional significance of these cerebral measures. PMID- 12558544 TI - Are patients with pituitary adenomas at an increased risk of mental disorders? AB - OBJECTIVE: Retrospective clinical studies suggest that patients with pituitary adenoma (PA) have an increased risk of mental disorders. However, empirical evidence for this hypothesis is deficient because of out-dated methodology. Therefore, we investigated the occurrence of mental disorders in 93 patients with PA within a limited time frame of 2 years. METHOD: Patients were subjected to a comprehensive, standardized lifetime and cross-sectional psychopathological symptom and diagnostic assessment, including an evaluation of psychosocial conditions and life events (CIDI, SCL-90) during the past years. RESULTS: We could not find an increased risk of mental disorders as compared with a representative population sample of 481 subjects matched by age and gender. CONCLUSION: As for our random sample, neither suffering from any type of PA nor the extent of the pituitary deficiency represent factors having significant effect on the risk of the subject to develop psychopathology as compared with the general population. PMID- 12558545 TI - Methylphenidate and intracortical excitability: opposite effects in healthy subjects and attention-deficit hyperactivity disorder. AB - OBJECTIVE: To test the effects of a standard dosage of the psychostimulant methylphenidate (MPH) - which significantly enhances intracortical inhibition but had no effects on intracortical facilitation in children with attention-deficit hyperactivity disorder (ADHD) - on intracortical excitability in healthy subjects. METHOD: In 12 healthy subjects, aged 20-40 years, intracortical inhibition and facilitation were investigated before and 70 min after the intake of 10 mg MPH using the technique of transcranial magnetic stimulation (TMS) with the paired-stimulus paradigm. RESULTS: In comparison of the two TMS measurements, a significant enhancement in intracortical facilitation but no effects on intracortical inhibition could be stated under MPH administration. CONCLUSION: This study provides first evidence for opposite effects of MPH on intracortical excitability in healthy adult subjects showing enhanced intracortical facilitation in contrast to ADHD children in whom enhanced intracortical inhibition has recently been shown. PMID- 12558546 TI - Asperger's disorder: a case report of repeated stealing and the collecting behaviours of an adolescent patient. AB - OBJECTIVE: To discuss special behavioural problems shown by a patient with Asperger's disorder from adolescence onward. METHOD: The case and treatment of a 21-year-old male patient is described. RESULTS: A 21-year-old male developed obstinate stealing behaviours when he was 17 years old. He was regarded as a schizophrenic at first, and was suspected of kleptomania later. Asperger's disorder was diagnosed after we reconsidered the relationship between the schizoid psychopathy in childhood and the stealing behaviours which occurred in adolescence. CONCLUSION: A wide variety of bizarre behaviours and so-called borderline behaviours occur in late adolescence and adult life of patients with Asperger's disorder. But classic schizophrenia is very rare. Psychiatrists unacquainted with the clinical diagnosis/context may find it difficult to evaluate 'concrete', 'childish', or 'bizarre' symptoms in patients with Asperger's disorder, and thus are prone to misdiagnose them as having schizophrenia disorders or other similar disorders. PMID- 12558547 TI - Alcoholism, violence, and CSF HVA. Cerebrospinal fluid homovanillic acid. PMID- 12558549 TI - Kinetic model of osmosis through semipermeable and solute-permeable membranes. AB - The gas analogy of the van't Hoff equation for osmotic pressure deltapi = RT/V, where R is gas constant, T absolute temperature and V mole volume of water, remained unexplained for a century because of a few misconceptions: (1) Use of supported membranes prevented the recognition that osmotic forces exert no effect on the solid membrane. During osmotic flow frictional force of solvent within membrane channels equals osmotic kinetic force pi at the interface against the solution containing impermeant solute. (2) Retrograde diffusion of water is much less than osmotic flow even when dx in the gradient dc/dx approaches zero. (3) The gas analogy was thought to be accidental. Actually, the internal kinetic pressure is P = RT/V, because intermolecular forces cancel out at the liquid interface, just as within a gas. The kinetic osmotic pressure is the difference in solvent pressure across the interface: pi = RT/V-(RT/V)X1 = (RT/V)X2, where X1 and X2 are the mole fractions of water and impermeant solute, respectively. Integration gives pi = -(RT/V)lnX1, identical to the thermodynamic equation. This equation is correct up to 25 atmospheres, and up to 180 atmospheres by assuming that a sucrose molecule binds 4 and a glycerol molecule 2.5 water molecules. For solute-permeable membranes, the reflection coefficient sigma can be calculated by formulas proposed for ultrafiltration. Because the fraction (1-sigma) of solute concentration behaves as solvent, osmosis may well proceed against the chemical potential gradient for water. The analogy to an ideal gas applies because pi = (RT/V)lnX1 is the small difference between enormous internal solvent pressures. PMID- 12558550 TI - Amazing chloride channels: an overview. AB - AIM: This review describes molecular and functional properties of the following Cl- channels: the ClC family of voltage-dependent Cl- channels, the cAMP activated transmembrane conductance regulator (CFTR), Ca2+ activated Cl- channels (CaCC) and volume-regulated anion channels (VRAC). If structural data are available, their relationship with the function of Cl- channels will be discussed. We also describe shortly some recently discovered channels, including high conductance Cl- channels and the family of bestrophins. We illustrate the growing physiological importance of these channels in the plasma membrane and in intracellular membranes, including their involvement in transepithelial transport, pH regulation of intracellular organelles, regulation of excitability and volume regulation. Finally, we discuss the role of Cl- channels in various diseases and describe the pathological phenotypes observed in knockout mice models. PMID- 12558551 TI - Blood flow distribution during elevated intraperitoneal pressure in the rat. AB - AIM: Oliguria is seen during elevated intraperitoneal pressure, but the physiological mechanisms are not yet clarified. The purpose of the present study was to investigate the changes in renal function, cardiac output and distribution of systemic blood flow (BF) that occur in connection with an elevation of intra abdominal pressure (IAP) in a rat model by isotope-labelled microsphere technique. METHODS: A 5 or 10 mmHg IAP was created by CO2 insufflation and maintained for 90 min in anaesthetized and mechanically ventilated rats. Rats with normal IAP served as controls. Blood flow and cardiac output measurements by injection of isotope-labelled microspheres were conducted at three time points. Acid-base balance, urine output, glomerular filtration rate (GFR) and urinary excretion products were also followed. RESULTS: Glomerular filtration rate decreased [0.7-0.1 mL min(-1) g(-1) kidney weight (KW)] with elevated IAP, as did urine output (8.5-0.6 microL min(-1) g(-1) KW). Dramatic decreases were seen in renal excretion of sodium (by 97%), potassium (by 94%) and osmotic active substances (by 93%). Cardiac output was diminished by 54% at 5 mmHg and by 65% at 10 mmHg intraperitoneal pressure and systemic vascular resistance (SVR) was elevated threefold. CONCLUSION: Cardiac output, measured by microsphere technique, decreased during elevated intraperitoneal pressure by CO2 in anaesthetized rats, while SVR was elevated and renal excretory functions were decreased to a large extent. PMID- 12558552 TI - Transmitter characteristics of cutaneous, renal and skeletal muscle small arteries in the rat. AB - AIM: We studied transmitter characteristics of proximal and distal arteries supplying skin (saphenous artery and its medial tarsal branch), kidneys (terminal branches of renal artery and interlobar arteries) and skeletal muscle (proximal and distal sections of external sural artery). METHODS: Artery segments were mounted in an isometric myograph and intramural nerves were activated by electrical field stimulation. Adrenergic and purinergic components of the neurogenic response were blocked using phenoxybenzamine and alpha,beta-methylene adenosine triphosphate (mATP), respectively. RESULTS: Arteries from skin or kidney developed rapid and prominent neurogenic contractile responses, with half maximal amplitude reached within 5-15 s; responses in proximal vessels were greater than in distal vessels. Arteries from skeletal muscle responded to sympathetic stimulation with a moderate contraction developing over 1 min or more, the response of distal segments was greater than that of proximal segments. In skeletal muscle vessels the sympathetically evoked contraction was completely blocked by phenoxybenzamine, whereas in skin and renal vessels it was the combined effect of noradrenaline and adenosine triphosphate (ATP). Given alone, mATP did not change the magnitude of the response to nerve stimulation, but increased its latency and also potentiated the response to exogenous noradrenaline. In all vascular beds, distal vessels were more sensitive to noradrenaline and mATP. CONCLUSION: It thus appears that the noradrenaline/ATP ratio of the sympathetic vasoconstrictor response differs between vascular beds in a way that is consistent with known differences in the selective regulation of regional vascular resistance by the sympathetic nervous system. PMID- 12558553 TI - Renal and hormonal responses to isotonic saline infusion after 3 days' head-down tilt vs. supine and seated positions. AB - AIM: The study aimed to determine whether prolonged exposure to simulated microgravity produces a level of thoracic volume receptor loading similar to that seen in the upright position or immediately after lying down. METHODS: We used a cross-over design to compare responses to a saline infusion in eight healthy subjects during a 4-day, -6 degree head-down tilt (HDT) and in the acute seated and acute supine positions. RESULTS: The first 24 h of HDT were associated with greater urinary excretion of water and sodium (UV, UNaV) than seated and acute supine [cumulative UV, 3035 +/- 219, 2311 +/- 156 (P < 0.05), and 2448 +/- 182 mL (P < 0.05), respectively; cumulative UNaV, 256 +/- 19, 180 +/- 11 (P < 0.05), and 189 +/- 15 mmol (P < 0.05), respectively]. Haemoglobin and haematocrit were increased after 24 h and plasma volume decreased after 48 h of HDT (P < 0.05). With prolongation of HDT, UV and UNaV returned near the baseline values, and plasma atrial natriuretic factor (ANF) and renin values returned to acute seated levels; in acute supine, ANF values were higher and renin lower than in the two other positions. After a 30-min infusion of 20 mL kg(-1) isotonic saline on the fourth HDT day or during acute seated or acute supine, sodium excretion within 4 h was similar during HDT and acute seated (83 +/- 6 and 84 +/- 9 mmol, respectively) and greater during supine (104 +/- 8 mmol, P < 0.05). The renin decrease was greater in HDT and seated than in supine. The plasma ANF increase was greater during HDT than during supine; during seated, plasma ANF was unchanged. CONCLUSION: These data suggest that, after 4 days of HDT, thoracic volume receptor loading returns to the same level as in the seated position, leading to blunted responses to volume expansion as compared with the acute supine position. PMID- 12558554 TI - Vasopressin increases milk flow and milk fat concentration in the goat. AB - AIM: The aim of this study was to investigate whether physiological doses of vasopressin affect milk flow and milk composition in goats. METHODS: Six goats were given vasopressin intravenously at doses of 0.1 ng kg(-1) min(-1) (a low dose, VL), 0.3 ng kg(-1) min(-1) (a medium dose, VM) and 1.0 ng kg(-1) min(-1) (a high dose, VH), oxytocin at 10 ng kg(-1) min(-1) and 0.15 m NaCl during 90 min at random. RESULTS: Milk flow increased from 1.3 +/- 0.2 to 3.5 +/- 1.0 g min(-1) (VL), from 1.8 +/- 0.4 to 4.3 +/- 0.6 g min(-1) (VM) and from 1.5 +/- 0.2 to 7.9 +/- 0.9 g min(-1) (VH) during the first 30-min infusions (all P < 0.001), respectively, and then declined. The response to oxytocin corresponded to that of VH. Milk fat concentration increased from 2.1 +/- 0.1 to 3.3 +/- 0.4% during VH and from 2.8 +/- 0.6 to 4.1 +/- 0.6% during oxytocin (both P < 0.001), and tended to increase during VM and VL. The mammary blood flow of four goats was studied with ultrasound during VH and oxytocin infusion. Milk flow increased from 1.8 +/- 0.4 to 4.7 +/- 1.0 g min(-1) during VH and from 1.9 +/- 0.3 to 12.4 +/- 2.4 g min(-1) during oxytocin (both P < 0.001). Mammary blood flow decreased from 262 +/- 37 to 53 +/- 7 mL min(-1) (P < 0.001) during VH, but did not change during oxytocin. CONCLUSION: We conclude that vasopressin increases milk flow and milk fat concentration and suggest that vasopressin, like oxytocin, acts on the myoepithelial cells. PMID- 12558555 TI - Differential strain patterns of the human gastrocnemius aponeurosis and free tendon, in vivo. AB - AIM: The mechanical characteristics of the human free tendon and aponeurosis, in vivo, remains largely unknown. The present study evaluated the longitudinal displacement of the separate free Achilles tendon and distal (deep) aponeurosis of the medial gastrocnemius muscle during voluntary isometric contraction. METHODS: Ultrasonography-obtained displacement of the free tendon and tendon aponeurosis complex, electromyography of the gastrocnemius, soleus, and dorsiflexor muscles, and joint angular rotation were recorded during isometric plantarflexion (n = 5). Tendon cross-sectional area, moment arm and segment lengths (L(o)) were measured using magnetic resonance imaging. Tendon force was calculated from joint moments and tendon moment arm, and stress was obtained by dividing force by cross-sectional area. The difference between the free tendon and tendon-aponeurosis complex deformation yielded separate distal aponeurosis deformation. Longitudinal aponeurosis and tendon strain were obtained from the deformations normalized to segment lengths. RESULTS: At a common tendon force of 2641 +/- 306 N, the respective deformation and Lo were 5.85 +/- 0.85 and 74 +/- 0.8 mm for the free tendon and 2.12 +/- 0.64 and 145 +/- 1.3 mm for the distal aponeurosis, P < 0.05. Longitudinal strain was 8.0 +/- 1.2% for the tendon and 1.4 +/- 0.4% for the aponeurosis, P < 0.01. Stiffness and stored energy was 759 +/- 132 N mm(-1) and 6.14 +/- 1.89 J, respectively, for the free tendon. Cross sectional area of the Achilles tendon was 73 +/- 4 mm2, yielding a stress of 36.5 +/- 4.6 MPa and Young's modulus of 788 +/- 181 MPa. CONCLUSION: The free Achilles tendon demonstrates greater strain compared with that of the distal (deep) aponeurosis during voluntary isometric contraction, which suggests that separate functional roles may exist during in vivo force transmission. PMID- 12558556 TI - Myosin remodelling in the contracting A7r5 smooth muscle cell. AB - AIM: Using confocal microscopy and standard immunohistochemical techniques, changes in the structure of alpha-actin and beta-actin as well as the distribution of myosin II were studied in the contracting A7r5 smooth muscle cell. RESULTS: Prior to stimulation, each of the three proteins were incorporated into filamentous structures extending the length of the cell body. At 30 min after stimulation by phorbol 12, 13 dibutyrate (PDBu), the system of densely packed beta-actin fibres was retained. By comparison, alpha-actin and myosin were observed to undergo significant remodelling, characterized by loss in fibre structure and the formation of brightly fluorescing peripheral bodies. Co immunoprecipitation of alpha-actin and myosin II suggested an association between the proteins. Consistent with this observation, dual immunostaining for alpha actin and myosin revealed strong co-localization of the two proteins prior to stimulation. Following PDBu stimulation myosin II was observed to partially disassociate from alpha-actin structure but showed significant co-localization with alpha-actin filaments and peripheral bodies throughout the interval of contraction. The use of cytochalasin B to block actin polymerization or the selective dissolution of alpha-actin cable structure by thapsigargin produced similar patterns of change in alpha-actin structure and the localization of myosin II. CONCLUSION: The results support the concept of myosin liability and potential for remodelling. The results suggest that myosin undergoes extensive relocalization in association with alpha-actin remodelling which may be an important determinant of contractile function in the A7r5 smooth muscle cell. PMID- 12558557 TI - [Pathogenesis of osteoporosis in rheumatoid arthritis]. AB - Osteoporosis is a major clinical problem in rheumatoid arthritis. Patients with rheumatoid arthritis frequently not only present with juxta articular osteopenia and bone erosions but also with generalized axial and appendicular osteoporosis at sites distant from inflamed joints. The pathogenesis of bone loss in rheumatoid arthritis is multifactorial; disease activity certainly is a major determinant of bone mass. Further pathogenetic factors include effects of anti inflammatory therapies (in particular glucocorticoids), reduced mobility, estrogen and/or androgen deficiency. Recently, receptor activator of nuclear factor kappa B ligand (RANKL) and osteoprotegerin (OPG), a decoy receptor for receptor activator of nuclear factor kappa B ligand, were identified as central regulators of osteoclast recruitment and activation. Osteoprotegerin and receptor activator of nuclear factor kappa B ligand production is modulated by several cytokines, growth factors and hormones. In rheumatoid synovium both fibroblasts and activated T cells express receptor activator of nuclear factor kappa B ligand and thereby promote osteoclast recruitment and activation. Thus, osteoprotegerin and receptor activator of nuclear factor kappa B ligand appear to represent important molecular links between the immune system and bone metabolism in rheumatoid arthritis. PMID- 12558558 TI - Quality of blood pressure control and risk of cerebral bleeding in patients with oral anticoagulation. AB - BACKGROUND: We assessed how many patients on long-term oral anticoagulation (OAC) (i) suffer from arterial hypertension (AH), (ii) are aware of AH, (iii) need improvement of their therapy and (iv) suffer from cerebral bleeding. METHODS: Outpatients on long-term OAC were asked to measure blood pressure at least 4 times. Blood pressure was classified as normotensive if at least 75 % of all measurements were < 139/89 mm Hg; as mild/moderate AH if > 25 % of all measurements were 140 - 179 mm Hg systolic or 90 - 109 mm Hg diastolic; and as severe AH if > 25 % of all measurements were > 180 mm Hg systolic or > 110 mm Hg diastolic. Bleeding complications were registered. RESULTS: Of the 235 patients (108 female, 67 +/- 12 years), 80 % suffered from AH. Severe AH was present in 5 %. Only 56 % were aware of suffering from AH. An improvement of antihypertensive therapy was needed in 64 %. Over 225 days, only one cerebral bleeding occurred. Blood pressure was normotensive in 30 % with known AH. CONCLUSIONS: Blood pressure control seems better in OAC patients than in normals, if the patient is aware of AH. Patients with AH on OAC are not aware of AH in > 50 %. Repeated blood pressure measurements in OAC are recommended, even if patients are not aware of AH. PMID- 12558559 TI - Palliative treatment of unresectable bile duct tumours. AB - PURPOSE: Report on outcome of intraluminal high-dose-rate iridium-192 (HDR-Ir192) brachytherapy with or without external radiotherapy in patients with unresectable bile duct tumours suffering from symptoms of malignant obstructive jaundice. MATERIAL AND METHODS: Fourteen patients (mean age: 63 years) who were unsuitable for surgical resection on preoperative evaluation/laparotomy or inoperable due to poor general condition were referred for palliative radiotherapy. After percutaneous transhepatic drainage, HDR-Ir192 brachytherapy was performed with a single dose of 2.5 Gy. Brachytherapy was given twice a day with at least a 6-h interval for 2 days, 2 or 3 days apart, up to a total dose of 10 Gy. Five patients received small-volume external radiotherapy (RT) (45 - 50.4 Gy/1.8 Gy) additionally. RESULTS: Palliation with relief of the aggravating symptoms of obstructive jaundice was achieved in all patients. The actuarial 2-year survival rate of all patients was 11.9 % with a median survival of 6.5 months. Patients treated with brachytherapy alone had a median survival of 4.5 months as compared with 6.5 months after combined internal and external irradiation (log rank, P = 0.95). CONCLUSION: Patients with advanced unresectable bile duct cancer face a dismal prognosis; however, biliary drainage, and intraluminal brachytherapy with or without external RT, seem to be able to improve quality of life in the remaining time span. PMID- 12558560 TI - Comparison of colonoscopies performed under sedation with propofol or with midazolam or without sedation. AB - BACKGROUND: Previous studies have shown that some patients are not able to tolerate colonoscopy without sedation because of low pain threshold, anxiety, colonic sensitivity and anatomical variations. Benzodiazepines are most commonly used, often in combination with pethidine. Our study compares sedation with propofol to midazolam and colonoscopy without sedation. METHODS: In this study 147 patients were examined. The patients were divided into three groups: The first group included patients in whom propofol was used for sedation, the second group included patients sedated with midazolam, and the third group was comprised of patients who received no sedation. Oxygen saturation and heart rate were recorded continuously by pulse oxymetry. Arterial blood pressure (BP) was monitored at 3-min intervals. RESULTS: A decrease in blood pressure and heart rate was documented in the first two groups (P < 0.001), whereas in group 3 we found an increase in both BP and heart rate (P < 0.001). Oxygen saturation dropped below 90 % in 11/102 patients sedated with propofol and in 9/23 sedated with midazolam. Two of 22 patients in whom no sedation was used were found to have oxygen saturation below 90 %. The recovery time was shorter (7 min) in group 1 (propofol) than in group 2 (midazolam), in which it was found to be 20 min. CONCLUSIONS: Our results showed that propofol provided good sedation with excellent pain control, a short recovery time and no significant haemodynamic side effects. PMID- 12558561 TI - 18F-FDG-PET in the follow-up of thyroid cancer. AB - Differentiated thyroid cancer is a rare tumour with an incidence of 4 - 9/100,000/year. For preoperative assessment of thyroid nodules, ultrasonography (US) and US-guided fine needle aspiration biopsy are the methods of choice to detect thyroid cancer. The value of preoperative fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG-PET) in differentiating malignant from benign nodules, especially in cases of follicular proliferation, has not yet been evaluated. After thyroidectomy and radioiodine remnant ablation, several methods are used to follow patients with differentiated thyroid cancer, including serum thyroglobulin, ultrasonography of the neck, iodine-131 (131I) whole body scintigraphy (WBS) and scintigraphy with nonspecific tracers such as technetium 99 m ((99m)Tc) Tetrofosmin or Sestamibi. Whereas the specificity of 131I-WBS is high, sensitivity is low, especially if one takes into account that only two thirds of recurrences or metastases store iodine. With the introduction of 18F FDG in oncology, it is also used for the detection of local recurrences and metastases of differentiated thyroid cancer. Elevated thyroglobulin but negative 131I-WBS belongs to the 1a indications for 18F-FDG-PET in oncology according to the German Consensus Conference 2000. The sensitivity for detecting 131I-negative metastases with 18F-FDG-PET can be increased by elevated thyroid-stimulating hormone (TSH) after withdrawal of thyroid hormone therapy or after intramuscular injection of recombinant TSH. Most of the 131I-negative metastases demonstrate 18F-FDG uptake, which represents rapid tumour growth and poor differentiation, whereas most of the 131I-positive metastases are 18F-FDG negative. The combination of 131I-WBS and 18F-FDG-PET leads to an increase in the detection rate to more than 90 - 95 % in cases of elevated thyroglobulin, because well- and less-differentiated cancer cells may be present in one patient. In rare cases, a recurrent tumour or metastasis may accumulate 131I as well as 18F-FDG. In these patients, it may be possible that well- and less-differentiated cells are present in one metastasis. The early use of 18F-FDG-PET in patients with elevated thyroglobulin, especially in the case of negative 131I-WBS, changes the therapeutic strategy in up to half of the patients (surgery, external radiation). PMID- 12558562 TI - Diagnosis in medullary thyroid cancer with [18F]FDG-PET and improvement using a combined PET/CT scanner. AB - Today, positron emission tomography (PET) investigation using fluorodeoxyglucose (FDG) is the most sensitive and specific single modality established for localizing metastases of medullary thyroid cancer (MTC). Even at calcitonin serum levels below 20 pg/ml, tumour localization may be successful in individual cases. However, especially in such early tumour stages, the morphological correlation using PET is unsatisfactory. With the newly available fixed combination of PET and computed tomography (PET/CT), this drawback can be overcome as PET/CT allows a point-by-point correlation of pathological function with morphological structure. Conventional PET alone yielded an approx 80 % sensitivity and specificity in MTC: a result that will be surpassed by the combined PET/CT scanner. PMID- 12558563 TI - Immature teratoma in both mediastinum and liver of a 21-Year-old female patient. AB - We present a case of an immature teratoma of the liver and the mediastinum. A 21 year-old female patient presented with dyspnea, abdominal pain, and abdominal mass. Abdominal ultrasonography showed a liver tumor which is located in the right lobe and composed of cystic and solid elements. Computed tomography (CT) also showed a big mass which contained multiple high density, nodular, semi solid, heterogenic structures in the liver and another mass which contained multiple low density cystic lesions in the anterior mediastinum. Right thoraco abdominal approach was performed on the patient and a complete macroscopic resection of tumors was achieved without complications. Both tumours were evaluated as having an intact capsule. The mediastinal tumour extended to the diaphragm on the right side, and is dissected from the 'surrounding lung, pericardium' and excised from diaphragm. Right hepatic lobectomy was performed for liver tumour. Intraoperative frozen sections of the liver tumor revealed teratoma. But the final pathological diagnosis was immature teratoma. Chemotherapy was given after surgery. The patient tolerated the procedure well and her postoperative course was unremarkable. At the first follow up 4 months after surgery she was alive and well and there was no evidence of recurrence, but the patient died within seven months with hepatic recurrence and spreading metastasis. PMID- 12558564 TI - Congo red-positive cardiac kappa-AL amyloidosis in plasmacytoma -- case report and review of the literature. AB - We report on a 51-year-old female patient who was diagnosed in 2001 as suffering from plasmacytoma. The patient had had complaints of bilateral carpal tunnel syndrome in 1999, treated by a simple dissection without performing histological examination. Congestive heart failure had gradually developed since that time. In 2001 echocardiography revealed a pronounced thickening of the left ventricular wall with systolic and diastolic dysfunction. A kappa-light chain M component and kappa-light chain-restricted bone marrow plasmacytosis were detected. Amyloid deposits staining positive in the kappa-light chain-restriction analysis were observed in a gastric biopsy. Taking into consideration all these findings, a plasmacytoma-associated systemic AL amyloidosis was diagnosed. Cyclophosphamide/prednisolone chemotherapy regimen led to complete haematological remission. Cardiac transplantation, combined with autologous peripheral blood stem cell graft, was considered as the next therapeutic step, but the patient died while on the waiting list for transplantation. Autopsy detected a highly hypertrophic myocardium with narrowed heart cavities. Microscopic examination revealed dense, pink, acellular, Congo red-staining and kappa-immunoperoxidase positive AL amyloid masses splitting the cardiomyocytes. The present case is remarkable as it demonstrates that carpal tunnel syndrome and congestive heart failure could be symptoms of plasma cell dyscrasia-associated amyloidoses. PMID- 12558565 TI - Determinants of health-related quality of life in adults with psychogenic nonepileptic seizures: are there implications for clinical practice? PMID- 12558566 TI - Differential expression of GABA and glutamate-receptor subunits and enzymes involved in GABA metabolism between electrophysiologically identified hippocampal CA1 pyramidal cells and interneurons. AB - PURPOSE: The balance between synaptic excitation and inhibition within the hippocampus is critical for maintaining normal hippocampal function. Even mild reduction in inhibition or enhancement of excitation can produce seizures. Synaptic excitation is produced by pyramidal cells and granule cells, whereas inhibition is produced by a smaller number of interneurons. To understand how two subpopulations of these excitatory and inhibitory neurons are regulated at the molecular level, we analyzed specific mRNA expression profiles for receptors that are significantly involved in synaptic transmission and in the synthesis and storage of the principal inhibitory neurotransmitter, gamma-aminobutyric acid (GABA). Our hypothesis was that differences in gene expression between inhibitory and excitatory neurons in the rat hippocampus might point to specific new targets for seizure pharmacotherapy. METHODS: We combined the techniques of (a) whole cell patch clamping in rat hippocampal slices, (b) biocytin staining for cell identification, (c) single-cell mRNA amplification, and (d) small-scale cDNA microarray analysis to allow us to obtain expression profiles for candidate genes from identified CA1 pyramidal neurons and interneurons. Electrophysiologic and morphologic data and expression profiles were obtained from 12 stratum pyramidale and seven stratum radiatum cells. RESULTS: Presumed inhibitory neurons expressed significantly more GAD65, GAD67, vGAT, GABA(A)-receptor alpha3, and N-methyl-d aspartate (NMDA)-receptor IIB mRNA, and presumed excitatory neurons expressed more GABA(A)-receptor alpha1, and NMDA-receptor I mRNA. CONCLUSIONS: Differential expression of candidate neurotransmitter-receptor subunits distinguished CA1 pyramidal neurons from interneurons. These differences may indicate potential new targets for altering the balance of inhibition and excitation in the treatment of epilepsy. PMID- 12558567 TI - Gabapentin increases the hyperpolarization-activated cation current Ih in rat CA1 pyramidal cells. AB - PURPOSE: Gabapentin (GBP) is a commonly used drug in the treatment of partial seizures, but its mode of action is still unclear. The genesis of seizures in temporal lobe epilepsy is thought to be crucially influenced by intrinsic membrane properties. Because the Ih substantially contributes to the intrinsic membrane properties of neurons, the effects of GBP on the Ih were investigated in CA1 pyramidal cells of rat hippocampus. METHODS: CA1 pyramidal cells in hippocampal slices were examined by using the whole-cell patch-clamp technique. RESULTS: GBP increased the Ih amplitude in a concentration-dependent manner mainly by increasing the conductance, without significant changes in the activation properties or in the time course of Ih. The effects ranged from approximately 20% at 50 microM, approximately 25% at 75 microM, to approximately 35% at 100 microM GBP (at -110 mV). In the presence of intracellular cyclic adenosine monophosphate (cAMP), the effects of GBP on Ih were similar to those obtained in the absence of cAMP. CONCLUSIONS: These results suggest that GBP increases the Ih through a cAMP-independent mechanism. Because the applied GBP concentrations were in a clinically relevant range, the observed effect may contribute to the anticonvulsant action of GBP in partial seizures and may represent a new concept of how this anticonvulsant drug works. PMID- 12558568 TI - The effect of electrical stimulation of the subthalamic nucleus on seizures is frequency dependent. AB - PURPOSE: Animal studies and anecdotal human case reports have indicated that the subthalamic nucleus (STN) may be a site of anticonvulsant action. METHODS: We tested the hypothesis that continuous electrical stimulation of the STN inhibits seizures acutely. We determined the effects of three stimulation frequencies, 130 Hz, 260 Hz, and 800 Hz, on generalized clonic and tonic-clonic flurothyl seizures. Adult male rats were implanted with concentric bipolar stimulating electrodes in the STN bilaterally. After recovery, rats underwent flurothyl seizures to compare the effects of each stimulation frequency on seizure threshold. Rats were tested 4 times, twice in the stimulated condition, and twice in the unstimulated condition. The order of trials was random, except that stimulation trials alternated with control trials. Flurothyl seizure thresholds under each stimulation condition were compared with control values from the same animal. RESULTS: Bilateral stimulation of the STN at 130 Hz produced a significant increase in the seizure threshold for clonic flurothyl seizures, whereas stimulation at 260 Hz did not appear to have any effect on seizures. STN stimulation at 800 Hz significantly lowered seizure threshold for tonic-clonic seizures. CONCLUSIONS: We conclude that electrical stimulation of the STN can be anticonvulsant, but the effects appear to depend on the stimulation frequency and the type of seizure. PMID- 12558569 TI - Amygdala kindling in Proechimys guyannensis rat: an animal model of resistance to epilepsy. AB - PURPOSE: This study examined the effect of amygdala kindling development in Proechimys guayannensis rat, a common rodent of the Amazon basin. METHODS: Adult male P. guayannensis animals (n = 43) and adult male Wistar rats (n = 14) were submitted to electrical amygdala kindling. RESULTS: From 43 Proechimys rats submitted to the kindling process, only three animals reached stage 5 of kindling. During the kindling development (stages 4-5), these animals had behavioral alterations different from those observed in Wistar rats. A longer time spent in stages 1-3 and 5 and longer afterdischarge duration in stages 1-4 was observed in the Proechimys group compared with the Wistar group. The number of wet-dog shakes also was reduced in the Proechimys group during the kindling process. CONCLUSIONS: These findings suggest natural endogenous inhibitory mechanisms in this animal species. PMID- 12558570 TI - Levetiracetam, a new antiepileptic agent: lack of in vitro and in vivo pharmacokinetic interaction with valproic acid. AB - PURPOSE: The novel antiepileptic drug (AED) levetiracetam (LEV; Keppra) has a wide therapeutic index and pharmacokinetic characteristics predicting limited drug-interaction potential. It is indicated as an add-on treatment in patients with epilepsy, and thus coadministration with valproic acid (VPA) is likely. These studies were performed to determine whether coadministration of LEV with VPA might result in pharmacokinetic interactions. METHODS: In vitro assays were performed to characterize the transformation of LEV into its main in vivo metabolite UCB L057. The reaction was examined for its sensitivity to clinically relevant concentrations of VPA. An open-label, one-way, one-sequence crossover clinical trial was conducted in 16 healthy volunteers to assess further the possibility of any relevant pharmacokinetic interaction. RESULTS: Human whole blood and, to a lesser extent, human liver homogenates were demonstrated to hydrolyze LEV to UCB L057, its main metabolite. The reaction possibly involves type-B esterases and is not affected by 1 mM VPA (i.e., 166 microg/ml). Pharmacokinetic parameters of a single dose of LEV (1,500 mg) coadministered with steady-state concentrations of VPA (8 days of 500 mg, b.i.d.) did not differ significantly from the pharmacokinetics of LEV administered alone [area under the curve (AUC) of 397 and 400 microg/h/ml, respectively]. Furthermore, LEV did not affect the steady-state pharmacokinetics of VPA. CONCLUSIONS: These findings suggest the absence of a pharmacokinetic interaction between VPA and LEV during short-term administration, and suggest that dose adjustment is not required when these two drugs are given together. PMID- 12558571 TI - Cardiac asystole in epilepsy: clinical and neurophysiologic features. AB - PURPOSE: Cardiac asystole provoked by epileptic seizures is a rare but important complication in epilepsy and is supposed to be relevant to the pathogenesis of sudden unexplained death in epilepsy (SUDEP). We sought to determine the frequency of this complication in a population of patients with medically intractable epilepsy and to analyze the correlation between EEG, electrocardiogram (ECG), and clinical features obtained from long-term video-EEG monitoring. METHODS: Retrospective analysis of the clinical records of hospitalized patients from May 1992 to June 2001 who underwent long-term video /EEG monitoring. RESULTS: Of a total of 1,244 patients, five patients had cardiac asystole in the course of ictal events. In these patients, 11 asystolic events, between 4 and 60 s long in a total of 19 seizures, were registered. All seizures had a focal origin with simple partial seizures (n = 13), complex partial seizures (n = 4), and secondarily generalized seizures (n = 2). One patient showed the longest asystole ever reported (60 s) because of a seizure. Cardiac asystole occurred in two patients with left-sided temporal lobe epilepsy (TLE) and in three patients with frontal lobe epilepsy (FLE; two left-sided, one bifrontal). Two patients reported previous cardiac disease, but only one had a pathologic ECG by the time of admission. Two patients had a simultaneous central ictal apnea during the asystole. None of the patients had ongoing deficits due to the asystole. CONCLUSIONS: These findings confirm that seizure-induced asystole is a rare complication. The event appeared only in focal epilepsies (frontal and temporal) with a lateralization to the left side. A newly diagnosed or known cardiac disorder could be a risk factor for ictal asystole. Abnormally long postictal periods with altered consciousness might point to reduced cerebral perfusion during the event because of ictal asystole. Central ictal apnea could be a frequent associated phenomenon. PMID- 12558572 TI - Interictal epileptic spiking during sleep and wakefulness in mesial temporal lobe epilepsy: a comparative study of scalp and foramen ovale electrodes. AB - PURPOSE: To assess distribution of temporal lobe spikes across different states of sleep and wakefulness in simultaneous scalp and foramen ovale (Fo) recordings. METHODS: The study included 12 patients with mesial temporal lobe epilepsy (MTLE). As part of their presurgical evaluation, patients underwent long-term video-EEG monitoring with combined scalp and foramen ovale electrodes (FoEs). In addition to traditional sleep scoring, waking was subdivided into eyes-opened and eyes-closed states, and rapid-eye-movement (REM) sleep was divided into phasic and tonic states. Spike counts were carried out visually for scalp and FoEs, and spiking rates were determined for each state. A ratio between FoE and scalp spiking rates also was calculated for each state. RESULTS: Scalp spiking showed a significant increase during NREM3,4, whereas FoE spiking increased during NREM2. The scalp/FoE ratio significantly increased during NREM3,4. A significant difference in spiking rate also was found between phasic and tonic REM states as well as between waking with eyes opened and closed in FoE recordings. CONCLUSIONS: Our data provide evidence of a discrepancy in spike distribution across different states of sleep and waking monitored by scalp and FoE recordings. We suggest that these discrepancies may reflect differences in archicortical and neocortical spike synchronization. PMID- 12558573 TI - Correlation of hippocampal glucose oxidation capacity and interictal FDG-PET in temporal lobe epilepsy. AB - PURPOSE: Interictal [18F]fluorodeoxyglucose (FDG) positron emission tomography (PET) demonstrates temporal hypometabolism in the epileptogenic zone of 60-90% of patients with temporal lobe epilepsy. The pathophysiology of this finding is still unknown. Several studies failed to show a correlation between hippocampal FDG-PET hypometabolism and neuronal cell loss. Because FDG is metabolized by hexokinase bound to the outer mitochondrial membrane, we correlated the glucose oxidation capacity of hippocampal subfields obtained after surgical resection with the corresponding hippocampal presurgical FDG-PET activity. METHODS: In 16 patients with electrophysiologically confirmed temporal lobe epilepsy, we used high-resolution respirometry to determine the basal and maximal glucose-oxidation rates in 400-microm-thick hippocampal subfields obtained after dissection of human hippocampal slices into the CA1 and CA3 pyramidal subfields and the dentate gyrus. RESULTS: We observed a correlation of the FDG-PET activity with the maximal glucose-oxidation rate of the CA3 pyramidal subfields (rp = 0.7, p = 0.003) but not for the regions CA1 and dentate gyrus. In accordance with previous studies, no correlation of the FDG-PET to the neuronal cell density of CA1, CA3, and dentate gyrus was found. CONCLUSIONS: The interictal hippocampal FDG-PET hypometabolism in patients with temporal lobe epilepsy is correlated to the glucose-oxidation capacity of the CA3 hippocampal subfield as result of impaired oxidative metabolism. PMID- 12558574 TI - Benign epilepsy with centro-temporal spikes: spike triggered fMRI shows somato sensory cortex activity. AB - OBJECTIVE: We performed spike triggered functional MRI (fMRI) in a 12 year old girl with Benign Epilepsy with Centro-temporal Spikes (BECTS) and left-sided spikes. Our aim was to demonstrate the cerebral origin of her interictal spikes. METHODS: EEG was recorded within the 3 Tesla MRI. Whole brain fMRI images were acquired, beginning 2-3 seconds after spikes. Baseline fMRI images were acquired when there were no spikes for 20 seconds. Image sets were compared with the Student's t-test. RESULTS: Ten spike and 20 baseline brain volumes were analysed. Focal activiation was seen in the inferior left sensorimotor cortex near the face area. The anterior cingulate was more active during baseline than spikes. CONCLUSIONS: Left sided epileptiform activity in this patient with BECTS is associated with fMRI activation in the left face region of the somatosensory cortex, which would be consistent with the facial sensorimotor involvement in BECT seizures. The presence of BOLD signal change in other regions raises the possibility that the scalp recorded field of this patient with BECTs may reflect electrical change in more than one brain region. PMID- 12558575 TI - Hippocampal region asymmetry assessed by 1H-MRS in rolandic epilepsy. AB - PURPOSE: In a previous study, we reported hippocampal abnormalities on magnetic resonance imaging (MRI) in six of 18 children with rolandic epilepsy (RE). In this study, metabolic changes were analyzed in the hippocampal region with proton magnetic resonance spectroscopy (1H-MRS). METHODS: In 13 children with electroclinically typical RE and 15 healthy controls, 1H-MRS results of both hippocampal regions were analyzed. The voxels, 2 x 2 x 4-cm each, were placed to include the head and body of the hippocampus. A PRESS sequence with TR 2,000 ms and TE 32 ms was used. Total N-acetylaspartate (tNAA), glutamine and glutamate (Glx), and choline compounds (tCho) were related to total creatine (tCr), and asymmetry indices (AIs) were calculated. MRI was performed in all 13 patients and in 13 controls. RESULTS: The tNAA/tCr AI of the hippocampal region was significantly higher in children with RE than in control children (z = 4.49; p < 0.001). The AIs of Glx/tCr and tCho/tCr did not show a significant difference between the groups. Lateralization of the interictal epileptiform activity corresponded with the lower tNAA/tCr ratio in 10 of 13 patients. MRI revealed a hippocampal asymmetry in four of 13 in the RE group, three of them showed concordance between the lateralization of the lower tNAA/tCr ratio and the smaller hippocampus. In the control group, a subtle asymmetry in four of 13 children was found. CONCLUSIONS: A significant asymmetry of the hippocampal regions, measured by tNAA/tCr ratios, indicates an abnormal neuronal function in children with RE. PMID- 12558576 TI - Levetiracetam efficacy in refractory partial-onset seizures, especially after failed epilepsy surgery. AB - PURPOSE: We conducted a retrospective study to evaluate the efficacy of levetiracetam as adjunctive therapy in patients with localization-related epilepsy, and specifically in the subset of patients for whom epilepsy surgery failed. METHODS: Eighty-two patients with uncontrolled partial-onset seizures treated with levetiracetam were identified; epilepsy surgery had failed for 21 (25.6%; group I), and 61 (74.4%) had no prior surgery (group II). Group I and group II patients were comparable in age (mean, 40.7 vs. 41.5 years) and age at seizure onset (mean, 14.4 vs. 18.2 years). Patients who had >/=50% reduction in seizure frequency were considered responders; the remaining patients were considered nonresponders. RESULTS: In patients (group I) for whom surgery had failed, responder rate was 76.1% (16 of 21), including 10 (47.6%) patients who became seizure free. In nonsurgical patients (group II), responder rate was 34.3% (21 of 61), including nine (14.7%) patients who became seizure free. In group I, 11 (91.6%) of 12 temporal resection patients were responders, of whom eight were seizure free; of the remaining nine operated (extratemporal) patients, five (55.5%) were responders, and two were seizure free. In three responders, all in group I, a severe, delayed psychotic syndrome developed 4 to 9 months after levetiracetam introduction, leading to its discontinuation. CONCLUSIONS: These findings suggest that adjunctive levetiracetam therapy should be considered early after failed epilepsy surgery, especially after temporal resection, and may have implications for its use before surgical intervention. Patients should be under close psychiatric observation in this clinical setting. PMID- 12558577 TI - The influence of sulthiame on EEG in children with benign childhood epilepsy with centrotemporal spikes (BECTS). AB - PURPOSE: To evaluate the effects of sulthiame (Ospolot; STM) monotherapy compared with placebo on the EEG in children with benign childhood epilepsy with centrotemporal spikes (BECTS). METHODS: Sixty-six patients (aged 3-11 years) entered a 6-month double-blind trial and were randomized to either STM (n = 31) or placebo (n = 35). Clinical data and general results have been reported elsewhere (1). One-hundred seventy-nine sleep EEGs were recorded at screening and after 4 weeks, 3 months, and 6 months. EEGs were analyzed by a blind reviewer using a standard protocol for each EEG. This standard protocol collected data on general changes, specific epileptiform, and nonspecific focal and generalized changes. A classification system was defined depending on rating of pathologic EEG changes. Because of the higher number of treatment-failure events (i.e., seizures) in the placebo group, there was an increasing imbalance between the two groups regarding the number of recorded sleep EEGs over time (STM, 104; placebo, 74). A Wilcoxon-Mann-Whitney U test was used to describe differences in the grade of pathology during individual follow-up between the two groups. RESULTS: The sleep-EEG was found to be normalized in 21 patients treated with STM (12/21 transient) and in five patients treated with placebo (4/5 transient). In the STM group, the EEG showed a marked improvement during intraindividual course when comparing the classification of follow-up EEGs at each time point with the screening EEG. Comparable improvements were not observed in the placebo group (exact two-tailed p value at 4 weeks, p < 0.0001; at 3 months, p = 0.0010; and at 6 months, p < 0.0001). CONCLUSIONS: STM had marked effects on the EEG in BECTS, which led to normalization in the majority of the patients. Most of those whose EEGs were not normalized showed improvement in the grade of EEG pathology. Normalization persisted in >50% of patients during the investigation. Spontaneous normalization in the placebo group reflects the wide spectrum of individual courses, which must be considered when analyzing drug effects on EEG in BECTS. PMID- 12558578 TI - Tactile-evoked rolandic discharges: a benign finding? AB - PURPOSE: This retrospective study was undertaken to determine if patients having tactile-evoked rolandic discharges were a more "benign" patient population than those with spontaneously occurring nontactile rolandic discharges and to determine whether the presence of tactile-evoked rolandic discharges was a marker for the future development of epilepsy, as previously reported. METHODS: During this 8-year study, 304 patients were seen with rolandic discharges. These patients all had tactile stimulation of their hands and/or feet. They formed two groups: patients with spontaneous rolandic discharges that could not be evoked by tactile stimulation (NT) and patients with spontaneous rolandic discharges that could be enhanced with tactile stimulation (TE). Over a 14-month period, every patient had tactile stimulation of both hands and both feet, resulting in a third group of patients having rolandic discharges seen only with tapping (TO). RESULTS: Tactile-enhanced discharges constituted 38.2% of all rolandic discharges. Patients with TE and TO discharges had a higher incidence of normal development and intelligence, normal neurologic examinations, and a lower incidence of seizures and focal or generalized background abnormalities on their EEGs. Only one patient with normal background and no coexisting epileptiform abnormalities had partial motor seizures with corresponding contralateral central discharges. Only two patients who had no seizures at the time of their first EEG subsequently went on to develop seizures. Neither fit the pattern of the seizure disorder described in the literature. CONCLUSIONS: It is hypothesized that tactile-evoked rolandic discharges are a benign, age-related phenomenon, which do not represent a marker for the future development of epilepsy and are not the interictal electrographic correlate to an already existing seizure disorder. PMID- 12558579 TI - Herbal remedies, dietary supplements, and seizures. AB - The use of herbal remedies and dietary supplements is widespread throughout the world, and use may be increasing. These are taken for a wide range of perceived benefits, such as energy and memory enhancement and treatment of specific conditions. Individuals with and without epilepsy may use these substances and may not inform their treating physician unless specifically asked. Inquiry about herbal medicine and dietary supplement intake should now be part of routine clinical history taking. Anecdotal accounts suggest that some herbal substances may have anticonvulsant effect, but randomised double-blind controlled trails are lacking. Alternatively many herbals and dietary supplements may predispose to seizures in individuals without epilepsy and worsen seizure control in those with epilepsy. In this article, we review the potential anticonvulsant and proconvulsant effects of herbal remedies and dietary supplements and discuss the potential interaction between these herbal substances and conventional anticonvulsant medications. PMID- 12558580 TI - Quality of life in psychogenic nonepileptic seizures. AB - PURPOSE: Psychogenic nonepileptic seizures (PNESs) are events that alter or seem to alter the neurologic function and, in their appearance, resemble epileptic seizures (ESs). In patients with ESs the psychological and medical aspects of epilepsy greatly influence the health-related quality of life (HRQOL). The relation between these factors and PNESs is not well established. In this study, we compared HRQOL in patients with PNESs with that of patients with ESs. METHODS: We evaluated 105 patients admitted to the Epilepsy Monitoring Unit of University Hospital between January 20, 2001, and January 20, 2002. Only patients with the definite diagnosis of ESs or PNESs were analyzed (n = 85). Patients completed an epilepsy-specific quality-of-life instrument (QOLIE-89), the Profile of Mood States (POMS), and Adverse Events Profile (AEP). We used t tests and regression analyses to contrast HRQOL in PNESs and ESs and to elucidate the main factors associated with HRQOL in patients with PNESs. RESULTS: In our sample, 45 patients had PNESs, and 40 had ESs. The overall HRQOL and scores on 13 of 19 QOLIE-89 subscales were significantly lower (i.e., worse) in PNES than in ES patients. AEP and scores on five of six POMS subscales also were worse in PNES patients than in ES patients. PNES versus ES diagnosis, POMS depression/dejection, and AEP were significant predictors of HRQOL, jointly explaining 65% variation in HRQOL. The lower HRQOL in PNESs versus ESs was in part explained by depression and AEP. CONCLUSIONS: Patients with PNESs have a lower HRQOL and worse mood problems than do patients with ESs. This disadvantage is primarily due to depression and medication side effects, although these factors influence QOL in much the same way in PNES and ES patients. These baseline HRQOL data on patients with PNESs can be used to evaluate the effects of treatment in this patient population. PMID- 12558581 TI - Epilepsy-induced changes in signaling systems of human and rat postsynaptic densities. AB - PURPOSE: To study seizure-induced changes in signaling proteins present in postsynaptic densities (PSDs) isolated from human epileptic neocortex and from rat cortex in which seizures were induced by injection of kainic acid. METHODS: We performed Western blot analysis of signaling proteins in PSDs isolated from cortical tissue. RESULTS: Seizures induce a strong upregulation of TrkB, the receptor for brain-derived neurotrophic factor (BDNF), whereas components of the N-methyl-d-aspartate (NMDA)-receptor complex are downregulated in both human and rat PSDs. CONCLUSIONS: These data show that long-term changes in PSD composition occur as a consequence of epileptic seizure activity. PMID- 12558582 TI - MRI volumetric analysis in rasmussen encephalitis: a longitudinal study. AB - PURPOSE: Rasmussen encephalitis is a progressive inflammatory process with difficult-to-control focal or lateralized seizure activity, leading to hemispheric dysfunction and atrophy in advanced stages. Anatomic changes of atrophy may be subtle in earlier phases of the disease, and progressive changes on serial scans may be difficult to detect. We report a case of early-stage Rasmussen encephalitis with a relatively stable clinical course in whom we performed magnetic resonance imaging (MRI)-based volumetric analysis over an interval of 1 year, to assess for volumetric changes. METHODS: Volumetric analysis was performed on two successive MRI scans obtained at age 5 and 6 years, by using the CARDVIEWS program (J Cogn Neurosci, 1996). The images were segmented into gray- and white-matter structures according to signal intensity of their borders semiautomatically, with manual corrections. The cerebral cortex was further subdivided into smaller parcellation units according to anatomic landmarks identifiable on MRI. RESULTS: Stable left cerebral hemispheric atrophy and progressive atrophy in the left precentral gyrus, left inferior frontal gyrus, and left cerebellar atrophy were detected over the 1-year interval. CONCLUSIONS: Volumetric analysis enables early detection and quantification of anatomic changes, identification of focal involvement, and assists in determining the severity of disease and timing for surgical interventions such as hemispherectomy. PMID- 12558583 TI - EEG in adult-onset idiopathic generalized epilepsy. AB - PURPOSE: It remains controversial whether adult-onset idiopathic generalized epilepsy (IGE) is a distinct syndrome or a continuum among IGE syndromes. EEG is the only known biologic marker of IGE and helps differentiate many of its classic subsyndromes. In this study, we looked for the differences in the EEG findings of IGE of classic adolescent onset versus adult onset that may suggest syndromic heterogeneity. METHODS: Seventy-six patients (47 adolescent-onset IGE, 29 adult onset IGE) with a clinical and EEG diagnosis of IGE were included. We defined IGE with age at onset of 11-20 years as adolescent-onset IGE and age at onset of 20 years or after as adult-onset IGE. Patients with first-decade onset of seizures, delayed EEGs, and no EEG available for review were excluded. The first EEG was performed within 24 h of the seizure, and if negative, a sleep-deprived EEG was done. All EEGs were reviewed in detail with respect to the background activity and the generalized spike-wave (GSW) characteristic. RESULTS: EEGs (87; 56 adolescent-onset IGE, 31 adult-onset IGE) were systematically reviewed. Background was normal in all patients. The morphology, amplitude, duration, frequency, occurrence, or activation of the GSW pattern did not differ between these two groups. CONCLUSIONS: No differences of EEG features were found between the classic adolescent-onset and the adult-onset IGE. This supports the hypothesis that they share common biologic determinants and exist along a life long age spectrum of classic IGE. PMID- 12558584 TI - Olfactory auras in patients with temporal lobe epilepsy. AB - PURPOSE: To investigate olfactory auras in patients with temporal lobe epilepsy (TLE). METHODS: We reviewed medical records of 217 Chinese patients who underwent temporal lobectomy for medically intractable TLE between 1987 and 1998 in Taiwan. Patients with olfactory auras asked for detailed characteristics of their auras. RESULTS: In all, 12 (5.5%) patients had olfactory auras, seven men and five women. All patients except one described and characterized the unpleasant olfactory auras. Olfactory auras were usually combined with other auras, most frequently sensations of epigastric rising, nausea, and fear. Association with gustatory hallucination was uncommon, in only one patient. On neuroimaging study, 11 patients had structural lesions involving the mesial temporal structures, two exclusively involving the amygdala. Histologic diagnosis included gliosis of the mesial temporal regions in seven (58.3%) patients, neoplasm in four (33.3%) patients, and arteriovenous malformation in one patient. Postoperatively, eight patients were seizure free. Three patients had rare seizures; however, none reported residual olfactory auras. CONCLUSIONS: Olfactory auras are infrequent in TLE. In this study, mesial temporal sclerosis is the most common etiology rather than tumors. Mesial temporal structures, especially the amygdala, may play important roles in the genesis of olfactory auras. PMID- 12558585 TI - Widespread upregulation of drug-resistance proteins in fatal human status epilepticus. AB - PURPOSE: Accumulating evidence implicates drug-transporter proteins ABCB1 and ABCC1 in resistance to multiple antiepileptic drugs (AEDs) in refractory epilepsy. These proteins are upregulated in surgically resected human brain tissue containing epileptogenic pathologies, including cortical dysplasia. In surgically resected cases, no upregulation is seen in normal adjacent tissue, suggesting that neither seizures nor prolonged exposure to AEDs need induce ABCB1 or ABCC1 expression. We wished to determine if status epilepticus might cause upregulation of these proteins. METHODS: Immunohistochemistry was performed for ABCB1 and ABCC1 in postmortem human brain tissue from a patient who died in status epilepticus and was found to have unihemispheric cortical dysplasia. RESULTS: There was upregulation of both proteins, as expected, in the hemisphere containing dysplasia. There also was widespread upregulation of both proteins in glia from the normal hemisphere. Previous work shows that drug treatment does not cause such upregulation. CONCLUSIONS: Upregulation of these proteins in histologically normal brain tissue is most likely the result of seizures in status, as seen in animal models. The findings provide a possible explanation for the appearance of AED resistance in prolonged status and emphasise the importance of prompt treatment of status epilepticus. PMID- 12558586 TI - Treatment of phenytoin toxicity by the molecular adsorbents recirculating system (MARS). AB - PURPOSE: Toxicity is common in patients of epilepsy treated with phenytoin (PHT), requiring careful drug level monitoring and supportive care. Specific treatment options are limited, although charcoal haemofiltration has been used previously. We attempted to demonstrate that severe PHT toxicity can be treated successfully with the Molecular Adsorbents Recirculating System (MARS). The mechanism of drug removal by the system also was studied. METHODS: A 45-year-old patient of status epilepticus with acute renal failure and severe PHT toxicity, associated with cardiac arrhythmias, hepatotoxicity, and altered sensorium, was treated with the MARS, a blood-purification system based on albumin dialysis, and including a charcoal filter, for 11.5 h. Serum PHT levels and blood levels of oxygen-based free radicals (by electron paramagnetic resonance spectroscopy) were measured before and after treatment. RESULTS: Serum total and free PHT levels declined sharply (32 to 11 microM and 9.8 to 2.0 microM, respectively), with clinical improvement and a 65% reduction in measured oxidative stress. The mechanism of drug removal, deduced by measuring PHT in the dialysate collected from different segments of the MARS circuit, was by clearance from blood into the albumin dialysate, and ultimately removal by the charcoal filter. CONCLUSIONS: The observed removal of PHT by MARS, along with the clinical improvement of the patient and reduction of the associated oxidative stress after treatment, indicates that MARS offers a promising option in PHT toxicity. PMID- 12558587 TI - Valproate-induced hyperammonemic encephalopathy. PMID- 12558589 TI - Survey of public awareness, attitudes, and understanding toward epilepsy in Hong Kong. PMID- 12558590 TI - Persistence of Salmonella enteritidis phage type 4 in the environment and arthropod vectors on an empty free-range chicken farm. AB - The persistence of S. Enteritidis PT4 was studied on a free-range breeding chicken farm which had been depopulated following identification of the organism in breeding birds. The site was sampled periodically for 26 months after depopulation and the organism was found to persist in litter, dried faeces and feed, but not in dust within empty poultry houses, for the whole of that period. Salmonella Enteritidis PT4 was also found in soil samples after 8 months but not 13 months and in faeces from wild mice, foxes and cats but not wild birds or badgers. The organism was also found in adult and larval forms of ground beetles and centipedes. Addition of pullets to a contaminated pen or inclusion of contaminated litter, feed or beetles/larvae to feed did not result in acquisition of infection by birds. PMID- 12558591 TI - Characterization of the surface hydrophobicity of filamentous fungi. AB - A method for the quantitative analysis of the hydrophobicity of the mycelial mat of filamentous fungi based on contact angle measurements is presented. It was tested for a range of fungi belonging to the classes of basidiomycetes, ascomycetes and deuteromycetes. The measured contact angles of the mycelial mats ranged between hydrophilic (<30 degrees) for the deuteromycetes Fusarium oxysporum Fo47 GUS1 and Trichoderma harzianum P1[pZEGA1] and hydrophobic (>60 degrees) for the ascomycete Cladosporium sp. DSE48.1b and the basidiomycetes Paxillus involutus WSL 37.7, Hebeloma crustiliniforme WSL 6.2, Suillus bovinus WSL 48.1 and Laccaria bicolor WSL 73.1. For some fungi, variations in the hydrophobicity of the mycelium depending on the growth medium, the physiological state and the exposure to water were distinguished. PMID- 12558592 TI - Physiological and molecular characterization of anaerobic benzene-degrading mixed cultures. AB - Nine distinct anaerobic benzene-degrading cultures were enriched from sediment samples from four different sites. These cultures used nitrate, sulphate or CO2 as electron acceptors. The shortest doubling times were observed in nitrate reducing cultures, although cell yield was lowest in these cultures. The highest substrate concentration utilized and maximum absolute rates of benzene degraded (in micro M day-1) were observed in methanogenic cultures. The microbial compositions of a methanogenic and nitrate-reducing culture were determined from a clone library of 16S rRNA genes. Five Bacterial 16S rRNA sequences, one of which resembled a clone previously found in a sulphate-reducing, benzene degrading culture and four Archaeal 16S rRNA sequences were identified in a methanogenic culture. Four Bacterial and no Archaeal 16S rRNA sequences were identified in a nitrate-reducing culture. The relative abundance of the four nitrate-reducing putative species was determined by slot blot hybridization. Two green sulphur bacteria together formed 52% of the clone library, but were found to be less than 4% of the culture by slot blot analysis. One of the cloned 16S rRNA gene sequences comprised 70% of the culture and was phylogenetically 93% similar to both Azoarcus and Dechloromonas species, which have been shown to degrade aromatic compounds, including benzene, under nitrate-reducing conditions. PMID- 12558593 TI - Persistence and cell culturability of biocontrol strain Pseudomonas fluorescens CHA0 under plough pan conditions in soil and influence of the anaerobic regulator gene anr. AB - Certain fluorescent pseudomonads can protect plants from soil-borne pathogens, and it is important to understand how these biocontrol agents survive in soil. The persistence of the biocontrol strain Pseudomonas fluorescens CHA0-Rif under plough pan conditions was assessed in non-sterile soil microcosms by counting total cells (immunofluorescence microscopy), intact cells (BacLight membrane permeability test), viable cells (Kogure's substrate-responsiveness test) and culturable cells (colony counts on selective plates) of the inoculant. Viable but non-culturable cells of CHA0-Rif (106 cells g-1 soil) were found in flooded microcosms amended with fermentable organic matter, in which the soil redox potential was low (plough pan conditions), in agreement with previous observations of plough pan samples from a field inoculated with CHA0-Rif. However, viable but non-culturable cells were not found in unamended flooded, amended unflooded or unamended unflooded (i.e. control) microcosms, suggesting that such cells resulted from exposure of CHA0-Rif to a combination of low redox potential and oxygen limitation in soil. CHA0-Rif is strictly aerobic. Its anaerobic regulator ANR is activated by low oxygen concentrations and it controls production of the biocontrol metabolite hydrogen cyanide under microaerophilic conditions. Under plough pan conditions, an anr-deficient mutant of CHA0-Rif and its complemented derivative displayed the same persistence pattern as CHA0-Rif, indicating that anr was not implicated in the formation of viable but non culturable cells of this strain at the plough pan. PMID- 12558594 TI - Intracellular symbionts of sharpshooters (Insecta: Hemiptera: Cicadellinae) form a distinct clade with a small genome. AB - The leafhoppers (Insecta: Hemiptera: Cicadellidae) are the most species-rich group of invertebrates in which intracellular symbionts are usual. Here we present the first molecular characterization of bacteriome-associates in the leafhoppers, with focus on the subfamily Cicadellinae (sharpshooters). Phylogenetic analyses of 16S rDNA sequences from intracellular symbionts residing in the bacteriomes of five host species indicate that these symbionts form a well defined clade within the gamma-3 Proteobacteria, consistent with an ancient colonization and strict vertical transmission. More extensive gene sequence information is reported for the symbiont of Homalodisca coagulata (Say). The genome size, as determined by pulsed field gel electrophoresis, is approximately 680 kb. This finding, when combined with published results for symbionts of aphids, ants, psyllids and tsetse flies, adds to an emerging pattern which suggests that bacteriome associates often descend from ancient infections by gamma Proteobacteria, and that these lineages have undergone pronounced genome reduction. A new genus and species name, 'Candidatus Baumannia cicadellinicola' (sp. nov.) is proposed for this newly characterized clade of symbiotic bacteria. PMID- 12558595 TI - A novel assay to monitor predator-prey interactions for Bdellovibrio bacteriovorus 109 J reveals a role for methyl-accepting chemotaxis proteins in predation. AB - Bdellovibrio bacteriovorus are Gram-negative bacteria that prey upon other Gram negative bacteria, including some pathogens, in a wide variety of habitats including soil, sewage, marine and estuarine environments. In order to facilitate studies on predation by this organism, we have developed a method that assays killing of luminescent Escherichia coli by B. bacteriovorus. Moreover, we have used this assay to compare predation of cells by derivatives of B. bacteriovorus containing targeted mutations in genes we have identified. Two genes are described; one, mcp2, encoding a methyl-accepting chemotaxis protein (MCP) and the other, an mviN homologue. Bdellovibrio bacteriovorus mcp2::aphII were less efficient predators on luminescent E. coli than B. bacteriovorus containing a randomly inserted aphII gene via TnphoA transposition. These and other chemotaxis experiments implicated at least a minor role for chemotaxis in predation by B. bacteriovorus. They also open the way for further studies on Bdellovibrio ecology, genomics and predator-prey interactions. The results further confirm that Bdellovibrio uses a chemotaxis system in order to sense, and respond to, changes in its environment, including prey. PMID- 12558596 TI - Testing the limits of biological tolerance to arsenic in a fungus isolated from the River Tinto. AB - The Tinto river in Spain, with its high acidity and heavy metal concentrations (As, Cu, Cr, Zn), is an example of an environment hostile to life. Yet despite these extreme conditions, the site possesses a great diversity of eukaryotic life forms. We report the isolation of a filamentous fungus able to grow at 200 mM arsenic ( approximately 15 000 p.p.m.), i.e. a concentration 20-fold above that withstood by the reference microorganisms Escherichia coli, Saccharomyces cerevisiae and Aspergillus nidulans, and 200 times greater than that tolerated by Aspergillus niger. Based on morphological, physiological and genotypic criteria, the strain belongs to the genus Aspergillus. High concentrations of the metalloid induced vacuolation, suggesting that this organelle is someway connected to arsenic tolerance. Concentrations that are lethal to other organisms do not stress Aspergillus sp. P37. The fungus was capable of removing arsenic from culture media. In addition to arsenic hyper-resistance, it also displayed a polyresistant phenotype to copper and chromium. PMID- 12558599 TI - Mouse embryonic mammogenesis as a model for the molecular regulation of pattern formation. AB - In this review we describe how mouse embryonic mammogenesis depends on a continuous communication between the epithelial and mesenchymal compartment of the mammary rudiment. Although the functions of only a few genes in the regulation of these epithelio-mesenchymal interactions during mouse mammary development are known so far, key roles are suggested for WNT, FGF and PTHrP signaling. However, the exact mechanism of action of these signaling pathways and their possible cross-talk in the induction of mammary development are not clear, nor does our current knowledge suffice to explain how the number and positions of the mammary rudiments are so well defined. Nonetheless, by the description of aberrant induction and/or maintenance of the mammary rudiments in a variety of inbred mouse strains and mutants, we have accumulated data demonstrating that the mammary rudiments develop independently of each other at these positions. In addition, each rudiment pair responds differently to altered levels of gene expression. This not only clarifies the unique identity of each placode, but the different molecular requirement of each placode also suggests that different molecular mechanisms may underlie the formation of such identical structures. For future investigations in the field, such a unique molecular identity of each mammary rudiment should be of critical concern. PMID- 12558600 TI - In vitro differentiation of embryonic stem cells into mineralized osteoblasts. AB - Embryonic stem cells are pluripotent cells derived from the inner cell mass of mouse blastocysts that have been shown to differentiate spontaneously into cell types representing all three germ layers. This study shows that ES cells were induced to differentiate in vitro into mineralized osteoblasts under the influence of ascorbic acid, beta-glycerophosphate and 1alpha,25-OH vitamin D3. The activity of alkaline phosphatase, an early osteoblast marker, was found to be increased around day 12 of culture. Mineralized cells were clearly identified by histochemical staining, which detects mineralized calcium. The major noncollagenous component of bone matrix, osteocalcin, was localized to the mineralized cells by immunofluorescence. The expression of bone-specific genes was analyzed by real-time quantitative PCR. Osteocalcin and bone sialoprotein (BSP) were identified as early as in the fourth week of embryonic stem cell culture, both being characteristic for late stages of osteoblastic differentiation, indicating that at this time of culture the identified cells represent "mature" osteoblasts. The osteoblast-specific transcription factor Cbfa1 was induced a few days earlier. The expression of osteopontin and osteonectin, both being involved in binding calcium ions and hydroxyapatite during mineralization processes, as well as of collagen type I, representing by far the most predominant collagen in vertebrate organisms, is enhanced at the beginning of the second culture week upon addition of supplements. In the third week of culture, treated cells showed a second peak of osteopontin, osteonectin and collagen type I expression, osteopontin and osteonectin being stimulated 3-4 fold and collagen type I being induced 6-fold over control values. Alkaline phosphatase (ALP) expression was enhanced at the beginning of the third week of culture and was found to be increased again at later stages of culture at days 27 34. The in vitro differentiation of mouse embryonic stem cells into osteoblasts may provide a suitable model for studying the molecular processes of osteoblastic development in vivo. PMID- 12558601 TI - Identification of a putative intestinal stem cell and early lineage marker; musashi-1. AB - There are few reliable markers for adult stem cells and none for those of the intestinal epithelium. Previously, indirect experimental approaches have predicted stem cell position and numbers. The Musashi-1 (Msi-1) gene encodes an RNA binding protein associated with asymmetric divisions in neural progenitor cells. Two-day-old, adult, and 4.5 h, 1-, 2-, 4- and 12-day post-irradiation samples of BDF1 mouse small intestine, together with some samples of mouse colon were stained with a rat monoclonal antibody to Musashi-1 (14 H-1). Min ( + / - ) mice with small intestinal adenomas of varying sizes were also analysed. Samples of human small and large bowel were also studied but the antibody staining was weak. Musashi-1 expression was observed using immunohistochemistry in neonatal, adult, and regenerating crypts with a staining pattern consistent with the predicted number and distribution of early lineage cells including the functional stem cells in these situations. Early dysplastic crypts and adenomas were also strongly Musashi-1 positive. In situ hybridization studies showed similar expression patterns for the Musashi mRNA and real-time quantitative RT-PCR showed dramatically more Msi-1 mRNA expression in Min tumours compared with adjacent normal tissue. These observations suggest that Musashi-1 is a marker of stem and early lineage progenitor cells in murine intestinal tissue. PMID- 12558602 TI - p38 MAPK interacts with actin and modulates filament assembly during skeletal muscle differentiation. AB - Skeletal muscle differentiation is marked by enhanced myotube formation and increased cytoskeletal rearrangement. Actin, a cytoskeletal protein is involved in various cellular functions such as glucose transport, intracellular trafficking, cell shape, and coordinated cell movement in response to various extracellular signals. The present study reveals an association between actin and p38 MAPK only in differentiated myotubes, not in proliferating myoblasts. Actin filament disassembly caused by cytochalasinD can be reversed using the potent activator of p38 MAPK, anisomycin. Pretreatment of myotubes with anisomycin partially resisted the effect of cytochalasinD. However, inhibition of p38 MAPK completely abolished the anisomycin-mediated actin remodeling. Data suggests that p38 MAPK interacts with actin and modulates actin filament rearrangement in differentiated L6E9 skeletal muscle cells. PMID- 12558603 TI - cAMP-dependent protein kinase regulates Polysphondylium pallidum development. AB - In eukaryotic cells, the universal second messenger cAMP regulates various aspects of development and differentiation. The primary target for cAMP is the regulatory subunit of cAMP-dependent protein kinase A (PKA), which, upon cAMP binding, dissociates from the catalytic subunit and thus activates it. In the soil amoeba Dictyostelium discoideum, the function of PKA in growth, development and cell differentiation has been thoroughly investigated and substantial information is available. To obtain a more general view, we investigated the influence of PKA on development of the related species Polysphondylium pallidum. Cells were transformed to overexpress either a dominant negative mutant of the regulatory subunit (Rm) from Dictyostelium that cannot bind cAMP, or the catalytic subunit (PKA-C) from Dictyostelium. Cells overexpressing Rm rarely aggregated and the few multicellular structures developed slowly into very small fruiting bodies without branching of secondary sorogens, the prominent feature of Polysphondylium. Few round spores with reduced viability were formed. When mixed with wild-type cells and allowed to develop, the Rm cells were randomly distributed in aggregation streams, but were later found in the posterior region of the culminating slug or were left behind on the surface of the substratum. The PKA-C overexpressing cells exhibited precocious development and formed more aggregates of smaller size. Moreover, expression of PKA-C under the control of the prestalk-specific ecmB promoter of Dictyostelium leads to protrusions from aggregation streams. We conclude that Dictyostelium PKA subunits introduced into Polysphondylium cells are functional as signal components, indicating that a biochemically similar PKA mechanism works in Polysphondylium. PMID- 12558604 TI - Multi-stage analysis of differential gene expression in BALB/C mouse liver development by high-density microarrays. AB - The development of a complex organ such as the liver relies on precise temporal and spatial gene expression patterns during ontogenesis. The unique adult phenotype is a result of a cascade of transcriptional events that finally trigger gene expression in a liver-specific fashion. Development in mice starts at embryonic stage E8.5-9.5 with the expression of several genes typically associated with liver tissue. While the role of some genes and their expression is well studied, little is known about the complex expression pattern changes during embryonic and fetal liver development. High-density oligonucleotide microarrays, which allow simultaneous expression analysis of 12,488 mouse mRNA transcripts and EST sequences, were used to study the gene expression profiles in day 7.5 embryonic tissue, in micro-dissected fetal liver tissue from day 11.5 and day 13.5 embryos, and in adult liver. In pairwise comparisons of all stages, a total of 4242 distinct genes or ESTs were found to be differentially regulated. Cross-comparisons of data from all stages detected the highest number of differentially regulated genes in E11.5 fetal liver tissue versus adult liver (3063 genes) and the lowest number in E11.5 versus E13.5 fetal liver tissue (517 genes). Using adult liver as reference tissue, 212 genes were regulated exclusively in E7.5 embryonic tissue, 303 genes in E11.5 and 198 in E13.5 fetal liver tissue. Expression profiles of the 31 genes with significant regulation at all stages as well as of a number of known developmentally regulated genes were compared with published results and interpreted. The gene expression profiles detected by microarray hybridization were independently confirmed for selected genes by quantitative RT-PCR. Our data presented here suggest that a relatively small number of stage-specific genes exist, which may be of particular importance for liver development, growth and differentiation. Furthermore, the microarray approach led to the identification of a number of genes, which have not yet been associated with liver organogenesis and maturation. PMID- 12558605 TI - cDNA sequences of the authentic keratins 8 and 18 in zebrafish. AB - From the zebrafish Danio rerio, we have cDNA cloned and sequenced a novel type II and a novel type I keratin, termed DreK8 and DreK18, respectively. We identified DreK8/18 as the true orthologs of the human keratin pair K8/18 as follows: (i) MALDI-MS assignment to the biochemically identified K8 and K18 candidates that are co-expressed in simple epithelia and absent in epidermal keratinocytes; (ii) multiple sequence alignments and phylogenetic tree analysis, showing that DreK8, within the phylogenetic tree of type II keratins, forms a highly bootstrap supported branch together with K8 from goldfish and rainbow trout, whereas DreK18, within the phylogenetic tree of type I keratins, groups with the K18 sequences from all other vertebrates studied; (iii) presence of a conserved motif in the tail domain of DreK8 (VxKxxETxDGxxVSESSxV) that is typical for all hitherto sequenced K8 orthologs. Moreover, several zebrafish type II keratin sequences published by other authors have now been assigned to epidermal keratins, previously identified biochemically. PMID- 12558606 TI - Cyclic expression of esr9 gene in Xenopus presomitic mesoderm. AB - During somitogenesis, the cycling expression of members of the Notch signalling cascade is involved in a segmentation clock that regulates the periodic budding of somites in chicken, mouse, and zebrafish. In frog, genes with cycling expression in the presomitic mesoderm have not been reported. Here, we describe the expression of Xenopus esr9 and esr10, two new members of the Hairy/Enhancer of split related family of bHLH proteins. We show that they are expressed in a highly dynamic fashion, with their mRNA levels oscillating periodically in the presomitic mesoderm during somitogenesis. This dynamic expression is independent of de novo protein synthesis. Thus, expression of esr9 and esr10 is an indicator of the segmentation clock in the amphibian embryo. This confirms the evolutionary conservation of a molecular pathway involved in vertebrate segmentation clock. PMID- 12558607 TI - On- and off-responses of heart rate to exercise - relations to heart rate variability. AB - During physical exercise, heart rate (HR) increases by parasympathetic withdrawal and increase of sympathetic activity to the heart. HR variability (HRV) in time and frequency domains provides information about autonomic control of the cardiovascular system. Non-linear analysis using the Poincare plot method is able to reveal supplementary information about cardiac autonomic control. The aim of this study was to determine the association between HRV parameters, the initial increase of HR at the onset of exercise (on-response) and HR decrease in the recovery phase after acute exercise (off-response). HR was continuously monitored in 17 healthy male subjects (mean age: 20.3 +/- 0.2 (SEM) years) at rest (25 min supine; 5 min standing), during exercise (8 min of step test at 70% of maximal power output) and in the recovery phase (30 min supine). HRV analysis in time and frequency domains and evaluation of the Poincare plot measures (length, widths) were performed on selected segments of HR time series. HR on- and off-responses were quantified using an exponential curve fitting technique. The time constants T(on) and T(off), representing the rate of on- and off-responses to exercise, were computed. Postexercise HRV indices and time constant of on-response - T(on) to exercise were negatively correlated. From preexercise HRV indices, only Poincare plot parameters were correlated with T(on). No correlation between HRV indices and parameters of off-response was found. In conclusion, preexercise HRV parameters are not closely correlated with the rate of cardioacceleration at the onset of exercise and cannot predict the rate of HR recovery. On the other hand, postexercise HRV parameters are related to the rate of initial adjustment of HR to exercise referring to the importance of rapid HR on-response for a faster recovery after exercise. PMID- 12558608 TI - Portapres and differential oscillometric finger blood pressure changes during deep breathing test in the assessment of BRS index. AB - The aim of this study was to investigate changes in the finger blood pressure during a deep breathing test (DB) and to find out whether the mean blood pressure might be used as a substitute for the systolic pressure in calculations of baroreflex sensitivity from data derived from the DB test. Continuous beat-to beat finger pressure was recorded by the volume clamp method (Portapres model 2 monitor). In addition, the mean arterial pressure was recorded by the modified oscillometric method (UT9201 beat-to-beat monitor, University of Tartu, Estonia). Fifteen healthy volunteers, aged 25-56 years, were studied. The amplitude of respiration-linked oscillations in the Portapres systolic (Psyst) and mean blood pressure (Pmean) was 22.2 +/- 8.8 and 16.6 +/- 5.8 mmHg, respectively. There was no significant difference between the amplitudes of induced changes in Pmean recorded by the two devices: the amplitude of oscillations in the mean blood pressure recorded by the differential oscillometric monitor was 16.0 +/- 5.9 mmHg. The amplitude of oscillations in Psyst correlated significantly with the amplitude of oscillations in Pmean recorded either by Portapres or by UT9201 (r=0.95 and 0.98, respectively). The high correlation between the amplitudes of oscillations in mean and systolic blood pressure allows to conclude that mean arterial pressure changes during a DB test might be used instead of systolic pressure changes in calculation of the ratio of changes in pulse interval to changes in blood pressure, which is considered to reflect baroreflex sensitivity. PMID- 12558610 TI - Reduction of risk factors following lifestyle modification programme in subjects with type 2 (non-insulin dependent) diabetes mellitus. AB - Exercise and improved diet is known to be beneficial in the management of type 2 (non-insulin dependent) diabetes mellitus. In practice, however, it is difficult for patients to implement these changes unaided. We hypothesized that a lifestyle modification programme involving residential visits would result in beneficial effects on glycaemic control and lipid profile. Three hundred and four individuals with type 2 diabetes participated in a lifestyle modification programme, involving three residential visits (2 weeks, 1 week and one 3-day visit) spaced over 31 weeks. The subjects were all referred for treatment following repeated failure to achieve metabolic control in primary care settings. Participants received information and practical guidance regarding exercise training, nutrition, as well as stress management and psychological counselling. Clinical parameters were determined at each visit. After completion of the programme, subjects showed significant improvements in glycaemic control (P<0.0001). Oxygen uptake was significantly improved (P<0.0001) and blood pressure (P<0.0001), body mass index (P<0.0001) and serum cholesterol (P<0.001) was significantly reduced, while HDL cholesterol (P<0.05) was significantly increased. There were no changes in LDL cholesterol values. Subjects also reported increased well-being and reduced stress. In conclusion, a 31-week lifestyle modification programme results in marked improvements in glycaemic control, blood pressure and well-being in subjects with type 2 diabetes. Thus, this type of lifestyle modification programme is a powerful treatment option to reduce risk factors associated with diabetes and diabetic complications, even in patients who have not responded to conventional diabetic therapy. PMID- 12558609 TI - Preserved bronchial dilatation after salbutamol does not guarantee protection against bronchial hyperresponsiveness. AB - Racemic salbutamol, a beta2-adrenoceptor agonist used for dilatation of airways, has recently been shown to induce lessened relaxation of bronchial smooth muscle and partial loss of bronchoprotection, seen as increased hyperresponsiveness, after regular treatment. The racemate undergoes stereo-selective disposition, giving higher plasma levels of S-salbutamol than that of bronchodilating R salbutamol, thus raising S : R ratios after repeated administration. Our aim was to evaluate whether increased bronchial hyperresponsiveness (BHR) could be found even after 1 day of repeated salbutamol inhalations, with beta2-receptor-induced bronchial smooth muscle relaxation remaining and whether this would be associated with plasma levels of either enantiomer. Fifteen patients with stable asthma, aged 19-54 years, were included in a randomized, cross-over study. An indirect bronchial challenge method was used [voluntary isocapnic hyperventilation of cold air (IHCA)], and airway condition tested by means of impulse oscillometry. Racemic salbutamol was inhaled three times during a 6-h period. IHCA was performed and plasma concentrations of enantiomers were measured 4 h after the last dose. Tests were also performed without preceding drug treatment. beta2 Agonist-produced bronchial dilatation and protection persisted in the majority of the 15 patients 4 h after repeated inhalations of salbutamol during 1 day. In only two of the 15 patients we could trace increased BHR after salbutamol. Neither dilatation nor protection could be linked to plasma levels of either R- or S-salbutamol. The underlying mechanisms of BHR remain unknown and are dissociated from beta2-receptor-mediated dilatation. PMID- 12558611 TI - Simultaneous non-invasive assessment of arterial stiffness and haemodynamics - a validation study. AB - The purpose of the study was to estimate the reliability of whole-body impedance cardiography (ICGWB)-derived pulse wave velocity (PWV) and stroke volume index to pulse pressure (SI/PP) measurements. The repeatability and reproducibility of ICGWB parameters were also determined. Agreement between the impedance and Doppler ultrasound-based PWV measurements was estimated in 25 healthy subjects in two consecutive measurements. Impedance-derived SI/PP (SIICG/PP) estimates were compared with simultaneously measured SI/PP based on thermodilution (SITD/PP) and direct Fick (SIFICK/PP) methods in 30 surgical patients. PWV measured between the aortic arch and popliteal artery using the impedance technique with selective electrode configuration (PWVIS) agreed well with the Doppler ultrasound method (PWVDOPP), the bias (PWVDOPP - PWVIS) and precision (+/- SD of differences) being 0.00 and 0.79 m s-1, respectively. PWV derived from the whole-body and popliteal impedance plethysmograms (PWVICG) overestimated slightly PWVDOPP values. The repeatability value for PWVIS was excellent, being 0.54 m s-1. The reproducibility values for PWVDOPP and PWVIS were very similar (2.17 and 2.42 m s 1, respectively). Changes in PWVIS correlated strongly with changes in PWVDOPP (r=0.74; P<0.0001), indicating that both methods reflected the true physiological variation in PWV. The agreement between SIICG/PP and SITD/PP or SIFICK was almost identical to the agreement between the SITD/PP and SIFICK/PP. In conclusion,whole body impedance cardiography provides handy and reliable means of evaluating arterial stiffness on the basis of PWV and SI/PP simultaneously with conventional haemodynamic parameters. The method is highly repeatable and reproducible. PMID- 12558612 TI - A longitudinal study of maternal oxygen saturation during short-term submaximal exercise. AB - PURPOSE: Longitudinal prospective study before, during and after normal pregnancy of the effect of short-term submaximal exercise on maternal oxygen saturation. METHODS: Fourteen healthy women were recruited to the study before a planned pregnancy, and were followed seven times during the pregnancy and for up to 6 month after delivery. A submaximal bicycle exercise test with a target heart rate of 85% of the predicted age-adjusted maximum was performed. Maternal oxygen saturation was continuously recorded using a pulse oximeter. RESULTS: Maternal oxygen saturation at maximum work-load had increased significantly already at 8 weeks gestation compared with preconception levels, and remained at a significantly higher level until 29 weeks of gestation. Thereafter, the oxygen saturation continued to be higher even at 6 month postpartum. During the exercise test, the lowest saturation was found during the late recovery period, this remained unchanged before, during and after pregnancy. CONCLUSION: A pregnant woman responds to short-term exercise by increasing the oxygen saturation until 29 weeks. After that, the saturation level decreases but remains at a higher level even 6 month after delivery compared with preconception levels. PMID- 12558613 TI - Influence of work rate on the determinants of oxygen deficit during short-term submaximal exercise: implications for clinical research. AB - OBJECTIVE: The effect of increasing work rate was studied on the determinants of the oxygen deficit. METHODS: Exercise testing was performed on a treadmill and gas exchange was measured on a breath-by-breath basis. Eleven healthy subjects, aged 18-25 years, performed three square wave exercise tests of different intensity. Before exercise, gas exchange was measured at rest in the standing position for 3 min, followed by a 6-min square wave exercise test, randomly assigned at 4, 8 or 12% inclination. Immediately after exercise the recovery gas exchange was determined for 3 min. To calculate oxygen deficit, the oxygen uptake (O2) values at onset of exercise were subtracted from the steady-state value, the differences were cumulated and expressed as a percentage of the total oxygen cost for the 6-min exercise. RESULTS: The oxygen deficit increased significantly (P<0.001) with increasing work rate (6.1 +/- 1.4% for 4%, 8.4 +/- 2.1% for 8% and 9.4 +/- 1.7% at 12% inclination). This resulted from a somewhat slower increase of O2 at the onset of exercise at the highest work rate, reflected by a significantly higher time constant for O2 at 8 and 12% (24.6 +/- 7.3 s at 8% and 24.1 +/- 6.3 s at 12% versus 20.2 +/- 8.1 s at 4%). More importantly a significantly higher steady-state value for O2 was found at the highest exercise level, compared with the other exercise intensities. CONCLUSION: The higher oxygen deficit at the highest level of exercise is determined by a slower time constant and a higher asymptote value for O2. PMID- 12558614 TI - Analysis of endothelium-dependent vasodilation by use of the radial artery pulse wave obtained by applanation tonometry. AB - OBJECTIVES: To evaluate applanation tonometry as a method to obtain arterial pulse waves suitable for pulse wave analysis of the height of the diastolic inflection point (IP), and to use this technique to study endothelium-mediated vasodilation by evaluation of the contribution of nitric oxide (NO) to the reduction in the height of the IP induced by beta2-adrenergic stimulation. METHODS: The radial artery pulse waveform was recorded by applanation tonometry in young healthy subjects before and after interventions both locally in the forearm and systemically by different vasodilators and vasoconstrictors, and vasodilatation was analysed as a change in the height of the IP. The mechanism behind the reduction in the height of the IP induced by terbutaline was investigated by systemic interventions with both N(G)-monomethyl-l-arginine (l NMMA) and noradrenaline (NA). RESULTS: Applanation tonometry was a convenient method to obtain radial artery pulse waves of good quality. The reduction in IP was substantially more pronounced when vasodilators were given systemically than when given locally in the forearm, indicating that the effect was obtained through an effect on peripheral pulse wave reflection. Systemically given l-NMMA, but not NA, increased the IP (P<0.05). Systemically given l-NMMA also caused a more pronounced attenuation than NA of the reduction in IP following terbutaline injection (P<0.05). CONCLUSION: Changes in IP following beta2-adrenergic stimulation appears to be a measurement of pulse wave reflection mainly governed by NO. Applanation tonometry and pulse wave analysis is a minimally invasive method suitable to assess endothelium-dependent vasodilation in large-scale studies. PMID- 12558615 TI - Lower heart rate response to ergometry rowing than to treadmill running in older men. AB - For older people exercise intensity is often determined based on heart rate (HR) or the percentage of maximal HR (%HRmax). This study evaluated oxygen uptake (O2) and HR during ergometry rowing (combined arm and leg; sitting exercise) and treadmill running (leg; upright exercise) for 15 older people [age, (mean +/- SD) 62 +/- 3 years]. The HR was lower during ergometry rowing than during treadmill running at a blood lactate concentration of 4 mmol l-1 (151 +/- 4 beat min-1 versus 160 +/- 5 beat min-1, P<0.05) and at a maximal effort (171 +/- 7 beat min 1 versus 177 +/- 7 beat min-1, P<0.05). This was the case although the O2 was higher during ergometry rowing than during treamill running both at a blood lactate concentration of 4 mmol l-1 (3.0 +/- 0.4 l min-1 versus 2.7 +/- 0.4 l min 1, P<0.05) and at a maximal effort (3.4 +/- 0.4 l min-1 versus 3.1 +/- 0.3 l min 1, P<0.05]. %HRmax and %HR reserve were lower during ergometry rowing than during treadmill running. The results suggest that, in prescription of rowing for older people, the relation between HR and O2 for rowing and the attenuated HR response to rowing should be taken into consideration. PMID- 12558618 TI - Axillary hyperhidrosis: eccrine or apocrine? AB - We review the literature regarding axillary hyperhidrosis, discuss normal sweat gland function and postulate on the respective roles of the eccrine, apocrine and apo-eccrine glands in the pathophysiology of excessive axillary sweating. PMID- 12558619 TI - Primary cutaneous B-cell lymphomas. AB - B-cell lymphomas comprise up to 25% of primary cutaneous non-Hodgkin lymphoma (NHL). Two main classification systems are used, namely the European Organization for Research and Treatment of Cancer (EORTC) and the World Health Organization (WHO) classification of tumours. Indeed, there is much controversy surrounding which classification system to use, which in turn influences the interpretation of existing data and subsequently how we manage patients with these conditions. PMID- 12558621 TI - Five cases of cutaneous Rosai-Dorfman disease. AB - Rosai-Dorfman disease (RDD), previously known as sinus histiocytosis with massive lymphadenopathy, is a benign, idiopathic histiocytic proliferative disorder. It commonly affects lymph nodes, but any organ of the body may be involved. Histological findings include characteristic large, pale, histiocytic cells (Rosai-Dorfman cells) exhibiting cytophagocytosis. Immunohistochemically, these histiocytes are positive for S-100 protein and CD68, but stain negatively for CD1a. On electron microscopy, Birbeck granules are absent. RDD limited to the skin is rare, less than 30 cases having been reported to date. We present five further cases of purely cutaneous RDD. Three presented as solitary nodules and one as a large, well-circumscribed plaque. The fifth patient, who was HIV positive, had a rosacea-like facial eruption. PMID- 12558620 TI - Treatment of cutaneous leishmaniasis with either topical paromomycin or intralesional meglumine antimoniate. AB - Ninety-six patients with a clinical and parasitological diagnosis of cutaneous leishmaniasis were recruited to a comparative randomized clinical trial evaluating the efficacy of topical paromomycin vs. weekly intralesional injections of meglumine antimoniate. The patients were randomly divided into two treatment groups: one group was treated with topical paromomycin ointment and the other with intralesional meglumine antimoniate. Treatment was continued in both groups until complete recovery occurred (defined as healing in less than 2 months with no residual scar or relapse for up to 1 year post treatment). Treatment failure was defined as an increase in the number and size of pre-existing lesions or untoward side-effects. The maximum treatment period was 3 months. The patients were followed up for 1 year. The results showed that intralesional meglumine antimoniate led to 41.7% complete recovery, However, topical paromomycin gave a lower cure rate of 16.6% (P < 0.05). Treatment failure was observed in 39.7% of the group receiving intralesional meglumine and in 72.9% of those on topical paromomycin (P < 0.05). This study indicates that intralesional meglumine antimoniate is superior to topical paromomycin in the treatment of cutaneous leishmaniasis. PMID- 12558622 TI - Complete remission of metastatic clear cell sarcoma with DAV chemotherapy. AB - We report the first case of metastatic clear cell sarcoma with dramatic response to DAV treatment (DTIC + ACNU + VCR). Clear cell sarcoma of tendons and aponeuroses, or malignant melanoma of soft parts, is a rare tumour that occurs predominantly in the extremities of young adults. It tends to recur locally or metastasize and the prognosis is poor. Although the importance of surgery has been established, the role of adjuvant chemotherapy has yet to be determined. DAV should be considered as a first-line palliative treatment in disseminated disease as well as adjuvant therapy after surgery of primary clear cell sarcoma. PMID- 12558624 TI - Cutaneous metastasis to the scalp from squamous cell carcinoma of the cervix. AB - Carcinoma of the cervix is a common neoplasm. Early detection and improved treatment techniques have resulted in better control of cancer and longer survival, however, invasive and metastatic disease still occur. Cervical cancer usually spreads through direct local extension and via the lymphatics, haematogenous metastasis is relatively infrequent and cutaneous metastases are very rare. Metastasis to the scalp is extremely rare and only three cases of scalp metastasis from cervical cancer have been reported in the literature. We now report a patient with cervical cancer who presented with metastasis to this very unusual site. PMID- 12558623 TI - Postmenopausal frontal fibrosing alopecia. AB - Recently a new entity, postmenopausal frontal fibrosing alopecia, was added to the established subtypes of scarring alopecias affecting postmenopausal women. This condition is characterized by a progressive frontal hairline recession associated with scarring. We studied the clinical and histopathologic features in four women with this disorder. Of note, a history of bilateral oophorectomy in two of them appears to be a new association. All four cases had frontoparietal recession of the hairline and two of them also had loss of their eyebrows. None of our four patients had any mucous membrane or other skin lesions. Histological examination showed perifollicular fibrosis and lymphocytic inflammation around the isthmus and infundibular areas of the follicles. No effective treatments have emerged for this type of postmenopausal alopecia, but progression of the hair loss and scarring appears to be self-limiting. We believe that this condition is a distinct clinicopathological variant of lichen planopilaris. PMID- 12558625 TI - Remarkable remission of a follicular lymphoma treated with rituximab and polychemotherapy (CHOP). AB - We describe a 50-year-old female patient who developed extensive lymphomatous infiltrates on her forehead, scalp and face within a few months. Histology and immunohistochemistry of skin tumours revealed a CD20 positive follicular B-cell lymphoma. Subsequently, extracutaneous manifestations were detected by computed tomography scans and bone marrow biopsy. The patient suffered from a primary nodular malignant lymphoma with extraordinary cutaneous infiltration of the head. Therefore, combination treatment with a monoclonal antibody against the CD20 antigen, rituximab, and polychemotherapy (CHOP scheme) was administered every 3 weeks. After the second course of treatment a complete regression of cutaneous infiltrates was noticed. Follow-up biopsies on the forehead showed no evidence of CD20 positive lymphoma cells, now. Despite mild leucocytopaenia therapy was well tolerated. PMID- 12558626 TI - Calciphylaxis in a patient with alcoholic liver disease in the absence of renal failure. AB - Calciphylaxis is a rare, potentially life-threatening syndrome characterized by progressive microvascular and superficial soft tissue calcification, usually seen in patients with chronic renal failure. We describe an unusual case of calciphylaxis in a patient with alcoholic liver disease and normal renal function who responded well to conservative wound care. PMID- 12558627 TI - A case of Trichosporon cutaneum folliculitis and septicaemia. AB - Trichosporon cutaneum, a saprophytic fungal organism normally found in the soil, has been increasingly recognized as a cause of life-threatening systemic illness in immunosuppressed patients including those with leukaemia. Cutaneous involvement occurs in about 30% of patients with T. cutaneum septicaemia although disseminated folliculitis has not been described to our knowledge. We now describe a case of T. cutaneum follicultis due to fungaemia in a neutropenic patient. PMID- 12558628 TI - Systemic lupus erythematosus-associated anetoderma and anti-phospholipid antibodies. AB - Anetoderma is characterized by a loss of normal elastic tissue that presents clinically as localized areas of wrinkled or flaccid skin. We describe the case of a 30-year-old woman with systemic lupus erythematosus-associated anetoderma and positive anti-phospholipid antibodies. We discuss the possible role of these antibodies in the pathogenesis of anetoderma, and, when detected, the need to check for an associated anti-phospholipid syndrome in such patients. PMID- 12558629 TI - Post-menopausal frontal fibrosing alopecia. AB - Post-menopausal frontal fibrosing alopecia (PFFA) has become an increasingly recognized distinct clinical entity in recent years. Most cases have been reported from Australia; however, it seems likely that the condition is under recognized. PFFA has similarities to lichen planopilaris but is differentiated by a distinctive symmetrical fronto-temporal distribution and progressive course. We report two cases from the UK and review the literature. PMID- 12558630 TI - Giant condyloma of Buschke-Loewenstein in association with erythroderma. AB - The Buschke-Loewenstein tumour is regarded as a type of verrucous carcinoma occurring on anogenital mucosal surfaces. The tumour is locally invasive but displays a benign cytology and rarely metastasizes. It is associated with human papillomavirus types 6 and 11. We describe a case of Buschke-Loewenstein tumour occurring in a 61-year-old man which behaved in a locally aggressive manner and was associated with human papillomavirus type 16 and erythroderma which proved resistant to treatment. The patient refused surgery and therapy with interferon alpha was ineffective. Chemotherapy with systemic cisplatin and 5-fluorouracil produced a partial response before the patient succumbed from gram-negative septicaemia. PMID- 12558631 TI - Intertriginous granular parakeratosis responsive to potent topical corticosteroids. AB - Granular parakeratosis is a recently recognized disorder of keratinization that is confined to intertriginous body sites. The histological features are distinctive. Aetiology is unclear at present but factors such as friction, perspiration and chemical irritation may be relevant. We describe three cases of granular parakeratosis that were notable for their rapid response to potent topical corticosteroids. PMID- 12558632 TI - Type XVII collagen gene mutations in junctional epidermolysis bullosa and prospects for gene therapy. AB - Non-Herlitz junctional epidermolysis bullosa (nH-JEB) is caused predominantly by mutations leading to premature stop codons on both alleles of the type XVII collagen gene (COL17A1). The analysis of mutations in this gene has provided a means of correlating genotype with phenotype of nH-JEB patients. The phenotype of nH-JEB is characterized by generalized blistering of skin and mucous membranes with atrophic scarring and nail dystrophy. Atrophic alopecia is a distinct feature of nH-JEB patients, but one that is not associated with the severity of the disease at other sites. Enamel hypoplasia and pitting of the teeth are also characteristic for nH-JEB and can be used to facilitate the correct diagnosis in children with a blistering skin disease. Analysis of the biological consequences of mutations in the COL17A1 gene has shown that most patients lack type XVII collagen mRNA due to nonsense-mediated mRNA decay. Patients with these mutations can therefore be a target for corrective gene therapy using vectors coding for full-length type XVII collagen. Proof of principle for this approach has recently been demonstrated. The analysis of naturally occurring phenomena of gene correction in the COL17A1 gene provides evidence for other mechanisms of gene correction in genetic diseases. For example, exclusion of an exon carrying a mutation can lead to a milder phenotype of nH-JEB than predicted by the original mutation. In addition, we have gained data suggesting that COL17A1 exons harbouring pathogenic mutations can also be repaired by trans-splicing, i.e. aligning corrected RNA sequences to introns in the vicinity of faulty exons in the COL17A1 premtRNA. PMID- 12558633 TI - Topical application of NADH for the treatment of rosacea and contact dermatitis. AB - Among many important physiological functions played by NADH (the reduced form of beta-nicotinamide adenine dinucleotide) its antioxidative properties are remarkable. Acting directly as an antioxidant, NADH can effectively protect the cell and its membrane from destruction by free radicals. NADH can be stabilized as a suspension in hydrophobic ointments prepared in a way that prevents contact with atmosphere containing oxygen and water. We present the first report of NADH as a treatment for some inflammatory dermatoses. It was found that topical application of 1% NADH diluted in Vaseline ointment can be very effective in the treatment of rosacea and contact dermatitis. Since no adverse effects were observed, therapy with NADH can be viewed as a potential alternative to other established treatments. PMID- 12558634 TI - Oral mucosal keratinocytes express RANTES and ICAM-1, but not interleukin-8, in oral lichen planus and oral lichenoid reactions induced by amalgam fillings. AB - Oral lichen planus (OLP) is a chronic inflammatory disease of the oral mucosa characterized by a band-like accumulation of lymphocytes in the connective tissue adjacent to the basement membrane as well as intraepithelially. Amalgam fillings can induce oral lichenoid reactions (OLR) that are similar to OLP. The adhesion molecule ICAM-1 and the chemokines interleukin-8 and RANTES all play central roles in leucocyte trafficking. The aim of this study was to investigate the possible role of these molecules in the migration of leucocytes into the oral mucosa in OLP and OLR. Standard immunoperoxidase techniques were used to visualize the expression of ICAM-1, RANTES and interleukin-8 in frozen biopsy sections. ICAM-1 was expressed by endothelial cells, but not by keratinocytes, in normal oral mucosa. ICAM-1 was expressed by keratinocytes in 11 of 12 biopsies of OLP and in six of seven biopsies of OLR. In all of these cases ICAM-1 was also expressed by endothelial cells and leucocytes. Although not present in normal oral mucosa, RANTES was expressed by keratinocytes in 21 of 24 biopsies of OLP and in seven of seven cases of OLR. Interleukin-8 was not detected in any of the samples. The expression of ICAM-1 and RANTES by epithelial keratinocytes in the oral mucosa in OLP and OLR could be a key inflammatory mechanism in these diseases. PMID- 12558635 TI - Association between human demodicosis and HLA class I. AB - Demodex folliculorum and D. brevis are obligatory parasites in hair follicles and in pilosebaceous glands of human skin. Although most people are infested with these mites, only a small number develop the clinical symptoms of skin demodicosis. The objective of this study was to determine the association between HLA specificity and demodicosis. Twenty-five patients with human demodicosis and 150 controls were typed for HLA-A, B, Bw, and Cw using the microlymphocytotoxicity method. The immune response was evaluated by identifying membrane markers of different immune cells using monoclonal antibodies. An association between the frequency of HLA Cw2 and Cw4 haplotypes and human demodicosis was established. The risk of developing clinical symptoms of this disease is 5.0 times higher for people with the Cw2 phenotype and 3.1 times higher for those with the Cw4 haplotype. Individuals who have the HLA A2 phenotype are 2.9 times more resistant to demodicosis. A positive correlation between demodicosis and the haplotypes A3-Cw4, A3-Cw2, A3-B17, A3-B35 and B35-Cw4 was found. In addition, an association between Cw2 and Cw4 alleles in the phenotype of patients with demodicosis and a decrease in the number of natural killer cells was found. PMID- 12558636 TI - Recessive X-linked ichthyosis associated with hypertrophic pyloric stenosis: a chance occurrence? AB - The association of recessive X-linked ichthyosis (RXLI) and hypertrophic pyloric stenosis (HPS) has been considered to be due to a probable contiguous gene defect. However, there are several reports of patients with large deletions on both sides of the steroid sulphatase gene (responsible for RXL1) that show no signs of HPS. We report the third pedigree wherein RXL1 was associated with HPS. Apart from the proband, both diseases showed themselves as independent events in the family tree with ichthyosis present in two other individuals and HPS in three other relatives. We calculated the probability that both diseases occurred simultaneously in the index case as a chance occurrence as 1 : 40 (using the Independence principle of probability). We conclude that in our pedigree it is likely that these two rare diseases show an accidental and not a true genetic association. PMID- 12558637 TI - A novel homozygous nonsense deletion/insertion mutation in the keratin 14 gene (Y248X; 744delC/insAG) causes recessive epidermolysis bullosa simplex type Kobner. AB - We report the sixth case of a human keratin 14 'knockout' mutation resulting in recessive epidermolysis bullosa simplex (EBS). A novel, homozygous nonsense mutation resulting from a deletion/insertion mutation (744delC/insAG) leads to a premature termination codon in the KRT14 gene (Y248X). The patient suffers from generalized cutaneous blistering since birth, mild nail dystrophy, involvement of mucous membranes and multiple epidermolysis bullosa naevi. The clinical variability noted in K14-deficient EBS patients suggests phenotypic modulation by additional genetic and/or epigenetic factors. PMID- 12558638 TI - Identification of mutations in the COL7A1 gene in a proband with mild recessive dystrophic epidermolysis bullosa and aortic insufficiency. AB - We report the clinical and molecular findings in a patient with a mild form of recessive dystrophic epidermolysis bullosa and aortic insufficiency. To our knowledge, this is the first report of association between dystrophic epidermolysis bullosa and abnormalities of the aortic valve. Analysis of the COL7A1 gene has revealed two new mutations, a 20-bp duplication and a splice site mutation. PMID- 12558639 TI - Holiday hazards: common stings from New World visits. PMID- 12558640 TI - The Neil Smith Memorial Lecture: John Laws Milton. The Founder of St John's Hospital for Diseases of the Skin. AB - John Laws Milton, a 19th-Century surgeon, founded the St John's Hospital for Diseases of the Skin close to Leicester Square in London in 1863. This article reviews his single-minded effort to establish dermatology as a medical subspeciality and to create a proper skin diseases hospital. The review also details his many contributions to the medical and dermatological literature, including his insightful description of the pregnancy dermatosis, herpes (pemphigoid) gestationis. PMID- 12558641 TI - Cutaneous anthrax associated with facial palsy. PMID- 12558642 TI - Bullous scabies in an adult. PMID- 12558643 TI - Ulcerated and nodular lesions on the vulva in a 60-year-old female. PMID- 12558644 TI - Basosquamous cell carcinoma with massive cranial osteolysis. PMID- 12558645 TI - Allergic periocular contact dermatitis due to atropine. PMID- 12558646 TI - Carbamazepine-induced pemphigus. PMID- 12558647 TI - Phototherapy in nodular prurigo. PMID- 12558648 TI - An unusual variant of lichen nitidus. PMID- 12558649 TI - Case 1. Incontinentia pigmenti. PMID- 12558650 TI - Case 2. Bleomycin-induced flagellate dermatosis. PMID- 12558651 TI - Case 3. Pyoderma faciale (PF) (rosacea fulminans). PMID- 12558652 TI - Case 4. Transient symptomatic zinc deficiency in a breast-fed premature infant: an acrodermatitis enteropathica-like eruption. PMID- 12558653 TI - Childhood scabies mistaken for Langerhans cell histiocytosis. PMID- 12558654 TI - Handicaps and the handicapped: a nomenclature and classification of intrinsic handicaps, by M. Agerholm. PMID- 12558657 TI - Mathematical models of hierarchically structured cell populations under equilibrium with application to the epidermis. AB - There are three categories of keratinocytes in the germinative compartment of the epidermis - stem, transit-amplifying and post-mitotic. Their population structure is hierarchical. This means that stem cells differentiate into transit-amplifying cells which, after a few rounds of division, become post-mitotic cells. The cell processes of birth, differentiation, death and migration affect the composition and proliferation rate of the germinative compartment. These phenomena are quantified by various cell kinetic parameters. In this paper we derive equations that relate these parameters for different models of hierarchically structured cell populations in equilibrium. We include in the models asymmetric and symmetric division, variations in cell-cycle times, apoptosis and variation in the number of transit generations. We conclude that variation in cell-cycle times need only be considered if apoptosis is not negligible. If it is negligible, then only average cell-cycle times are needed. Unfortunately, it is impossible to predict the importance of apoptosis from the available experimental data. However, the strength of its effect is determined by the other parameters, especially the fraction of cycling stem cells. We show that variation in the number of transit generations can have a potentially large effect on cell birth rate. We also show that cell birth rate does not directly depend on the mean transit-amplifying cell-cycle time, only on the mean stem cell-cycle time. We argue that 'homogeneous cell population' equations should not be used to study hierarchical cell populations as has been done in the past. Finally we argue that stem cell parameters and transit-amplifying cell parameters should not be lumped together. PMID- 12558658 TI - The rate of apoptosis in post mitotic neutrophil precursors of normal and neutropenic humans. AB - Using data on the fraction of post-mitotic neutrophil precursors (CD15+ cells) displaying positive markers for apoptosis in 12 normal humans, and a simple mathematical model, we have estimated the apoptotic rate to be about 0.28/day in this compartment. This implies that the influx of myelocytes into the post mitotic compartment exceeds twice the granulocyte turnover rate (GTR), and that about 55% of the cells entering this compartment die before being released into the blood. The normal half life of apoptotic post-mitotic neutrophil precursors is calculated to be 10.4 h. Comparable calculations for patients indicate apoptosis rates in the post-mitotic compartment of about 17 times normal for one myelokathexis patient and rates of about 13 times normal for the one cyclical neutropenic patient and two severe congenital neutropenic patients. The estimated half life for apoptotic post-mitotic neutrophil precursors in the myelokathexis patient was about 0.4 h, 1.4 h in the cyclical neutropenia patient, and about 0.6 h in the severe congenital neutropenic patients. PMID- 12558659 TI - The role of various Bcl-2 domains in the anti-proliferative effect and modulation of cellular glutathione levels: a prominent role for the BH4 domain. AB - Reduced cell proliferation and increased levels of cellular glutathione (GSH) are characteristic for cells that overexpress the anti-apoptotic Bcl-2 protein. We investigated the influence of various Bcl-2 domains on both these characteristics. Rat CC531 colorectal cancer cells were stably transfected with the human bcl-2 gene (CCbcl2 cells) or with bcl-2 gene constructs missing a coding sequence for a func-tional domain, BH1 (CCDeltaBH1 cells), BH3 (CCDeltaBH3 cells), BH4 (CCDeltaBH4 cells) or the transmembrane region (CCDeltaTM cells). We measured GSH levels in exponentially and confluent growing bcl-2-transfected cell populations. The fraction of S-phase cells during exponential growth was significantly reduced in CCbcl2, CCDeltaBH1, CCDeltaBH3, and CCDeltaTM cells compared with parental CC531, neo-transfected CC531 and CCDeltaBH4 cells. GSH levels in these bcl-2 transfectants were significantly higher than in the parental line measured at 50% confluence; at 100% confluence they reached a similar level as found in parental cells. Independently from the presence of BH1, BH3 or TM domains, overexpression of Bcl-2 reduces cellular proliferation under conditions of increased GSH levels. This apparent link is lost in CCDeltaBH4 cells; these cells are not reduced in cellular proliferation and harbour significantly higher GSH levels than found in the other transfectants. Studies on the subcellular localization revealed an extremely low expression of the Bcl-2 protein lacking the N-terminal BH4 domain in nuclear fractions. Nuclear translocation of Bcl-2 requires the presence of the BH4 domain and seems prominent in reducing cellular proliferation. PMID- 12558660 TI - Overexpression of Raidd cDNA inhibits differentiation of mouse preadipocytes. AB - RAIDD (RIP-associated ICH-1 homologous protein with a death domain) is an adaptor molecule that mediates the action of cysteine proteases involved in apoptosis. To study the possibility of a novel system of cell ablation mediated by RAIDD, a preadipocyte cell line (3T3L1) was stably transfected with a plasmid containing the murine Raidd cDNA under the control of the adipocyte specific promoter aP2. Instead of the expected apoptosis, a blockage to differentiation upon hormonal induction was observed as judged by an absence of lipid accumulation, a lack of expression of adipocyte-specific genes and a fibroblastic appearance. Proliferation rate of Raidd-transfected clones remained unaffected. Overexpression of Raidd cDNA in 3T3L1 cell therefore inhibited differentiation, suggesting that Raidd plays a role in controlling differentiation of mouse preadipocytes and, perhaps, in other cell types, in addition to its established role in apoptosis. PMID- 12558661 TI - Sucrose inhibits vascular smooth muscle cell proliferation/migration--protein synthesis is maintained. AB - We have previously shown that the onset of smooth muscle cell proliferation in tissue cultures is triggered independently of serum. The aim of the present study was to investigate if this process was affected by osmotic stress. Vascular explants from 8-month-old male rats were cultured under serum-free conditions using collagen I as migration substrate. Sucrose was added to the culture medium in concentrations varying from 1 to 3% (30-90 mOsM). Cell migration from aortic explants onto the culture dishes was totally inhibited at a sucrose concentration of 90 mOsM. A significant dose-dependent decline in proliferation was shown for cells in explants pulse labelled with 3H-thymidine. In contrast, pulse labelling with 35S-methionine revealed that protein synthesis was maintained in the presence of sucrose. The results indicate that osmotic pressure affects smooth muscle cell protein synthesis, proliferation and migration. PMID- 12558662 TI - Does LCIS or ALH without other high-risk lesions diagnosed on core biopsy require surgical excision? PMID- 12558663 TI - The management of lobular neoplasia identified on percutaneous core breast biopsy. AB - The management of lobular neoplasia (LN) found on percutaneous core biopsy remains a clinical dilemma. The purpose of this study was to establish guidelines for the management of LN when obtained on percutaneous core needle biopsy. A retrospective review of the Breast Imaging Tissue Sampling Database at New York Presbyterian Hospital-Columbia Comprehensive Breast Center was performed from 1998 to 2000. A total of 1460 percutaneous core breast biopsies were performed using 11- or 14-gauge needles with LN identified in 43 biopsies from 34 patients. Eleven biopsies were ultrasound guided for nonpalpable masses and 32 were stereotactically guided for mammographically detected densities (10) and microcalcifications (22). The 43 LN biopsies were divided into three groups based on additional findings associated with LN on core biopsy: group I (n = 19), LN with invasive cancer or ductal carcinoma in situ (DCIS); group II (n = 11), LN plus a second indication for open surgical biopsy, such as atypical ductal hyperplasia (ADH), radial scar, phyllodes tumor, or intraductal papilloma; and group III (n = 13), LN plus benign fibrocystic changes. In group I, 19 of 19 biopsies (100%) yielded invasive cancer or DCIS on surgical biopsy versus 3 of 11 (27%) for group II, and 1 of 13 (8%) for group III. Outcomes in group III are described as follows: three patients were lost to follow-up, three patients did not undergo surgical biopsy but demonstrated more than 1 year of mammographic stability following core biopsy. Of the remaining seven patients, two had LN and ADH on surgical biopsy (one had a contralateral cancer), one had atypical lobular hyperplasia (with a contralateral cancer), two had LN and benign fibrocystic changes, one had LN and intraductal papilloma, and one had LN and invasive ductal carcinoma (IDC) with DCIS (with a contralateral cancer). These results suggest that surgical biopsy is indicated for patients with LN when found on core biopsy and when the biopsy demonstrates invasive cancer, DCIS, or other indications for surgical biopsy such as ADH, or in the examination of a patient with a synchronous contralateral breast cancer. The diagnosis of LN alone without these indications on percutaneous biopsy may not warrant routine surgical biopsy. PMID- 12558664 TI - The significance of atypical lobular hyperplasia at percutaneous breast biopsy. AB - Atypical lobular hyperplasia (ALH) is occasionally found in specimens obtained by percutaneous stereotactic vacuum-assisted breast biopsy for microcalcifications. Since malignancy is often found at surgical excision when atypical ductal hyperplasia is found at percutaneous biopsy, we reviewed our pathologic findings from surgery for ALH at percutaneous biopsy. This was a retrospective review of all percutaneous breast biopsy specimens for mammographic microcalcifications obtained from a single institution over a 30-month period. The pathologic findings from percutaneous biopsy were correlated with the radiologic appearance and the pathology from surgical excision. ALH was found in 13 of 766 (1.7%) stereotactic vacuum-assisted core needle biopsies performed for mammographic microcalcifications. Subsequent surgery in six patients revealed ductal carcinoma in situ (DCIS) in two patients and one case of invasive ductal carcinoma. Surgical excision is indicated for areas with ALH discovered by percutaneous biopsy for mammographic microcalcifications. PMID- 12558665 TI - Invasive lobular carcinoma: spectrum of enhancement and morphology on magnetic resonance imaging. AB - Invasive lobular carcinoma (ILC) may be a difficult tumor to detect early by physical examination, mammography, or ultrasound. We undertook this study to describe the spectrum of gadolinium enhancement and morphologic features of ILC on magnetic resonance imaging (MRI). Nineteen patients with ILC who presented with a palpable mass, a mammographically visible abnormality, or an unknown primary underwent preoperative MRI of both breasts using a T1-weighted high resolution gradient echo sequence (pre- and postcontrast), and an echoplanar sequence during the administration of gadolinium. Using a quantitative measure of gadolinium uptake over time, called the extraction flow (EF) product, and a normal tissue threshold EF level of 25 or less, enhancement for 15 of the 19 cancers was characterized. By consensus, three radiologists categorized the morphologic features of the lesions. For the 15 cases of ILC that had echoplanar data, analysis showed peak EFs ranging between 25 and 120, and the majority showed EFs in the 30s. A substantial portion of two tumors enhanced in a similar fashion to normal breast tissue, with EFs in the low 20s. Morphologically MRI showed a focal mass in eight cases, regional enhancement in five, segmental enhancement in one, segmental enhancement with multiple small nodules in one, a mixture of a focal mass and regional enhancement in one, diffuse enhancement in one, multiple small nodules in one, and bilateral disease in one. Of the focal masses, seven were irregular in shape and one was round; six had ill-defined margins and two had spiculated margins. All eight enhanced heterogeneously. Four cases had multifocal disease and one case had unsuspected contralateral disease discovered only on MRI. MRI using a combination of morphology and a quantitative measure of gadolinium uptake was able to detect the majority of cases of ILC. However, there was a variable morphologic appearance and contrast enhancement pattern on MRI. A few lesions were difficult to distinguish from normal tissue. This suggests that some cases of ILC may be difficult to detect on MRI. PMID- 12558666 TI - Prevalence of family history of breast and ovarian cancer in a single primary care practice using a self-administered questionnaire. AB - Women at high risk of hereditary breast and/or ovarian cancer require specific management strategies for cancer prevention and early detection. The goal of this study was to determine the prevalence of familial breast and ovarian cancer among patients in a primary care practice. Questionnaires were mailed to the 608 women less than 81 years of age in a single primary care practice. Additional mailings and phone calls were used for nonresponders. Data were analyzed by bloodline, the degree of relative, age of diagnosis and cancer type. Women were grouped into three categories of breast/ovarian family history: "no family history,""insignificant family history," and "significant potentially high-risk family history" (women with two or more relatives in a single bloodline with breast and/or ovarian cancer, a single individual with bilateral breast cancer or breast and ovarian cancer, or breast and/or ovarian cancer at less than 40 years of age). A pedigree analysis of women categorized as "significant potentially high-risk family history" further classified these women as to the likelihood of being at risk for hereditary cancer. Data were obtained from 567 women (93%); 27 patients with a personal diagnosis of breast and/or ovarian cancer were excluded. Of the 540 remaining respondents, 351 (65%) had no family history of cancer, 138 (25.6%) had an insignificant family history, and 51 (9.4%) had a significant family history. Based on pedigree analysis of these 51 patients, 19 were unlikely to be at high risk for hereditary cancer, and 32 (6%) were likely to be at significant risk and warrant intensive evaluation. The large proportion of women identified with a significant family history of breast and/or ovarian cancer has major implications regarding the magnitude of a population-based process to identify and manage high-risk individuals. PMID- 12558667 TI - Palpable breast carcinomas: a hypothesis for clinically relevant lymphatic drainage in sentinel lymph node biopsy. AB - Previous studies have shown that independent of tumor size, palpable breast tumors have a higher incidence of lymph node metastasis compared with nonpalpable tumors. This study further examines this phenomenon using a large sentinel lymph node (SLN) database. Data from a prospective, institutional review board (IRB) approved, multi-institutional study from the University of Louisville Breast Cancer Sentinel Lymph Node Study Group was used. From August 1997 through December 2001, 3192 patients with clinical T1 and T2 N0 breast cancer underwent SLN biopsy, most with a combined technique of radioactive colloid and blue dye, followed by level I/II axillary dissection. Patients with palpable tumors tended to be younger (mean age 58 years) compared with nonpalpable tumors (mean age 61 years). The incidence of positive axillary metastasis was significant between palpable and nonpalpable tumors (43% and 23%, respectively), independent of tumor size by logistic regression (p = 0.0001). The SLN identification rate was significantly different between palpable and nonpalpable tumors (95% versus 91%, respectively; p < 0.0001). A unifying theory to explain the phenomenon that palpable tumors, stage for stage, are associated with a higher rate of nodal metastasis is that palpable tumors are, on average, closer to the skin and the rich network of dermal lymphatics. We believe that the dermal lymphatics of the breast represent a clinically relevant metastatic pathway to the axilla. PMID- 12558668 TI - Location and extent of positive resection margins and ductal carcinoma in situ in lumpectomy specimens of ductal breast carcinoma examined with a microscopic three dimensional view. AB - The location of positive margins in lumpectomy specimens for ductal carcinoma could be predicted due to the common pattern of the geographic relationship between the intraductal and invasive carcinomas. To test this hypothesis, 62 lumpectomy specimens for ductal carcinoma of the breast were submitted for this study. The specimens were microscopically examined by serially sectioning them into giant sections in a plane parallel to the chest wall (frontal plane). The margins were identified as proximal (closest to the nipple), distal (opposite to proximal), and peripheral (nonproximal or distal). We found that the location of positive or close margins was proximal in 6 cases, peripheral in 13 cases, and none were found to be distal. Ductal carcinoma in situ (DCIS) was found to be located in the area adjacent to the invasive carcinoma. The invasive carcinoma was located at the periphery of the intraductal carcinoma. All six specimens with invasive carcinoma without DCIS had free margins. Nine of 16 specimens (56%) with extensive intraductal carcinoma (EIC) component and 7 of 40 (18%) with DCIS but negative EIC contained positive or close margins involved by DCIS. One case with multifocal invasive carcinoma measuring 3.5 cm in diameter and with DCIS but EIC negative had margins involved by both DCIS and invasive carcinoma. In conclusion, in ductal carcinoma, invasive carcinoma arose at the peripheral areas of the DCIS. DCIS tends to spread toward the nipple and the peripheral margins of the resected specimens. Incomplete excision of the ductal carcinoma and the wide positive margins are most likely caused by the failure to estimate the extent and location of DCIS. PMID- 12558669 TI - Atypical ductal hyperplasia presenting as pseudomicrocalcifications. AB - A 53-year-old white female on mammography had "suspicious" microcalcifications. Upon excision, these turned out to be pseudomicrocalcifications created by the intraductal cellular proliferations of atypical ductal hyperplasia so dense as to create radiodensities. PMID- 12558670 TI - Bilateral brachial plexus palsy after a right-side modified radical mastectomy with immediate TRAM flap reconstruction. AB - A 65-year-old women developed bilateral brachial plexus palsy after unilateral modified radical mastectomy with TRAM flap reconstruction. The patient recovered spontaneously. The failure of appropriate intra-operative positioning to prevent the injury is discussed. PMID- 12558671 TI - Lobular carcinoma of the breast metastatic to the epidural space: a potential mimic of giant cell bone tumor. AB - We report the case of a 64-year-old woman with a spinal epidural mass. Tissue from a decompression laminectomy disclosed a tumor with numerous osteoclast-like giant cells separated by small, moderately atypical tumor cells. The osteoclast like giant cells were immunoreactive for vimentin, but negative for epithelial membrane antigen and broad-pectrum cytokeratin. Subsequent breast biopsy revealed a lobular carcinoma of classic type without osteoclast-like giant cells. This is the first reported case in which metastatic breast carcinoma was accompanied by these giant cells but the giant cells were not present at the primary tumor site. PMID- 12558672 TI - Staining of endotracheal tube with isosulfan blue dye after sentinel node mapping: a case report. AB - Sentinel node mapping is successfully performed for the detection of cancer and nodal metastasis. There is a reported complication rate of 1.5%, most being mild allergic reactions, with a few rare reported cases of anaphylaxis. Isosulfan blue has been found to routinely create a modest artifactual desaturation via pulse oximetry, as well as a transient skin discoloration that may last for several hours to weeks. It causes discoloration of urine and has been detected in other body secretions, none of which have been found to be of clinical significance. This is a case report involving a 61-year-old woman who underwent sentinel node mapping and axillary dissection for breast adenocarcinoma. On postoperative extubation, the endotracheal tube was noted to be stained dark blue with isosulfan dye. This unusual complication is unreported to date and is likely the result of systemic absorption of the dye. PMID- 12558673 TI - Spontaneous unilateral nipple discharge: when screening tests are negative--a case report and review of current diagnostic management of a pathologic nipple discharge. AB - We describe a 45-year-old woman who presented with a spontaneous unilateral nipple discharge. With a negative breast examination and screening tests (mammography and ultrasonography) she underwent mammary ductography, which revealed a small 3-4 mm intraluminal filling defect. A core biopsy showed high grade ductal carcinoma in situ (DCIS). An attempted wide local excision was unsuccessful, and the patient underwent a mastectomy. Pathologic assessment revealed high-grade DCIS and multiple foci of invasive mucinous ductal adenocarcinoma. Rare tumor cells were identified in the subcapsular sinuses in both sentinel lymph nodes. We report this case to point out the importance of the diagnostic examination for patients with a pathologic nipple discharge and review current and possible future diagnostic management. PMID- 12558674 TI - Giant fibroadenoma. PMID- 12558675 TI - A 10-year delayed breast cancer patient with unusual widespread cutaneous involvement and visceral metastases. PMID- 12558676 TI - Benign axillary lymph node inclusions. PMID- 12558677 TI - Metastasis of breast carcinoma to rectus abdominis muscle. PMID- 12558678 TI - Multidisciplinary care for breast cancer: barriers and solutions. AB - Breast cancer treatment and research is becoming more multidisciplinary in nature. Several modalities and areas of expertise are critical for optimal patient management. The basis for medical decisions and recommendations must reflect outcomes and clinical trial data that are designed and interpreted with broad input across different fields. Hence there has been a trend for specialization in breast disease in many large community and academic practices. Furthermore, a system of communication and standardization of data values, procedures, and protocols has begun, but needs much further development. There are many natural barriers to the process of multidisciplinary care and research in terms of logistics, finances, and education. The example of preoperative neoadjuvant therapy for early stage locally advanced breast cancer is one that involves multiple disciplines in the formulation of a treatment and in future research that will define the optimal individualized approach. This process can also shed further light on biologic principles and potential for improved treatment. Solutions for overcoming barriers to multidisciplinary care should include incentives for collaborative and coordinated clinical care across disciplines. A model of increased efficiency because of pooled resources and specialization in several fields should also be accompanied by a demonstration of increased quality of care and patient satisfaction. Any process that adds to cost or inconvenience needs to be justified in an evidence-based manner. Finally, these initiatives need to be effectively communicated to the professional and policy-making communities and to the public at large through well-conceived and unbiased educational venues. PMID- 12558679 TI - Capecitabine and PPE syndrome: a case report. PMID- 12558681 TI - Editorial. PMID- 12558680 TI - Does evidence-based medicine provide evidence? The evidence on breast cancer and the safety of oral contraceptives suggests that unorthodox interpretations may be just as valid. PMID- 12558682 TI - Saving the photons: mapping X-rays by position-tagged spectrometry. AB - Since the early years of X-ray spectrometry in electron microscopes, mapping the locations of chemical elements has been important. The X-rays needed in large numbers for this are rare, owing to poor production efficiency compared with electron signals, and at risk of loss by many mechanisms such as missing the limited solid angle of the detector, absorption before reaching the detector and pulse pile-up conventional digital mapping hardware reduces the information contained in the X-ray spectrum at each pixel to the itegrated counts from a few regions of interest.The acquisition technique of position-tagged spectrometry eliminates the conflict between the desire to see full frame X-ray images quickly versus the analytical advantages of having complete spectra for each pixel. As the beam is scanned rapidly relative to traditional X-ray mapping, photons are counted in a virtual 3-D multichannel analyser on disk, preserving both spatial and spectral information. Along with the sophisticated post-processing allowed by storing an entire spectrum per pixel, a unique degree of dynamic interaction with the developing data is made possible by integrating many short scans instead of using a single long dwell time at each pixel. PMID- 12558683 TI - Determination of the inelastic mean free path of electrons in vitrified ice layers for on-line thickness measurements by zero-loss imaging. AB - The inelastic mean free path of 120 keV electrons in vitrified ice layers has been determined in an energy-filtering TEM. From the ratio of the unfiltered and zero-loss-filtered image intensities recorded with a slow-scan CCD camera, the relative sample thickness t/Lambda can be calculated. For calibration, the geometric ice thickness was measured by imaging a tilted view of a cylindrical hole which had been burnt into the ice layer. The total inelastic mean free path was found to be 161 nm, and the partial inelastic mean free path for an acceptance angle of 4.2 mrad was 232 nm. These results were built into a standard protocol for use in cryo-electron microscopy allowing on-line measurements of local ice-layer thicknesses by zero-loss-filtered/unfiltered imaging. PMID- 12558684 TI - En bloc optical sectioning of resin-embedded specimens using a confocal laser scanning microscope. AB - Reconstruction of 3D structures of specimens embedded for light or electron microscopy is usually achieved by cutting serial sections through the tissues, then assembling the images from each section to reconstruct the original structure or feature. This is both time-consuming and destructive, and may lead to areas of particular interest being missed. This paper describes a method of examining specimens which have been fixed in glutaraldehyde and embedded in epoxy resin, by utilising the autofluorescence preserved or enhanced by aldehyde fixation, and by using a confocal laser scanning microscope to section optically such specimens in the block down to a depth of about 200 &mgr;m. In this way, the accurate estimation of the depth of particular features could be used to facilitate subsequent sectioning at the light microscope or electron microscope level for more detailed studies, and 3D images of tissues/structures within the block could be easily prepared if required. PMID- 12558685 TI - Imaging streptavidin 2D crystals on biotinylated lipid monolayers at high resolution with the atomic force microscope. AB - Streptavidin crystals were grown on biotinylated lipid monolayers at an air/water interface and transferred onto highly oriented pyrolytic graphite (HOPG). These arrays could be imaged to a resolution below 1 nm using the atomic force microscope. The surface topographs obtained were compared with negative-stain electron microscopy images and the atomic model as determined by X-ray crystallography. The streptavidin tetramer (60 kDa) exposes two free biotin binding sites to the buffer solution, while two are occupied by linkage to the lipid monolayer. Therefore, the streptavidin 2D crystals can be used as nanoscale matrices for binding biotinylated compounds. Furthermore, this HOPG-based preparation method provides a general novel approach to study the structure of protein arrays assembled on lipid monolayers with the AFM. PMID- 12558687 TI - A comparison of image restoration approaches applied to three-dimensional confocal and wide-field fluorescence microscopy. AB - We have compared different image restoration approaches for fluorescence microscopy. The most widely used algorithms were classified with a Bayesian theory according to the assumed noise model and the type of regularization imposed. We considered both Gaussian and Poisson models for the noise in combination with Tikhonov regularization, entropy regularization, Good's roughness and without regularization (maximum likelihood estimation). Simulations of fluorescence confocal imaging were used to examine the different noise models and regularization approaches using the mean squared error criterion. The assumption of a Gaussian noise model yielded only slightly higher errors than the Poisson model. Good's roughness was the best choice for the regularization. Furthermore, we compared simulated confocal and wide-field data. In general, restored confocal data are superior to restored wide-field data, but given sufficient higher signal level for the wide-field data the restoration result may rival confocal data in quality. Finally, a visual comparison of experimental confocal and wide-field data is presented. PMID- 12558689 TI - High angle [001] twist boundaries in platelet colonies of BiSrCaCuO (2223) superconductor. AB - The microstructure of BiPbSrCaCuO (2223) ceramic was studied by polarised light microscopy and the etch-pits technique. The high-angle twist grain boundaries in the platelet colonies were confirmed to be [001] coincidence type twisted boundaries or rotation twin boundaries with the (001) twinning plane. PMID- 12558688 TI - Photogrammetric calibration of a stereo light microscope. AB - Quantitative stereo light microscopy has been used in very rare cases to obtain geometric representations of microscopic objects. Although the instrument itself has limitations, most of them efficiently overcome by other 3D microscopes, it bears considerable advantages in observing and measuring dynamic scenes with multiple objects. The biggest asset of the stereo method is that the full 3D work space is imaged in one shot - a property which distinguishes stereo from all the scanning techniques in 3D microscopy. With the work presented in this paper, we contribute to making the stereo technique applicable to the numerous mensuration tasks in the microscopic domain, where its potential would be invaluable.We report the photogrammetric calibration of a common main objective lens type stereo light microscope. Such a calibration is the initial step in ensuring accurate measurements with this instrument. First, we derive a mathematical formulation of the imaging function and discuss the estimation of the parameters involved. Then, three main problems of the practical implementation of the framework are addressed: the finding of a calibration standard, the automatic measuring of the many image coordinates required, and the stabilization of the parameter estimation. Finally, results of various calibration runs are presented and analysed under different aspects. Among these, the most important is the accuracy of the calibrated instrument in measuring 3D positions and positional relationships. With a Zeiss Stemi 11, Achromat 1.6 x we achieve accuracies of 1 per thousand laterally and 1-2% axially relative to the volume of the work space. On the highest magnification level this corresponds to 700 nm and 1.8 &mgr;m, respectively. PMID- 12558690 TI - Assessment and diagnosis of depression in people with intellectual disability. AB - BACKGROUND: Despite widespread acceptance that depression can occur in adults with intellectual disability (ID), the difficulties encountered in its assessment and diagnosis have hampered the individual clinician, and meant that questions of prevalence, treatment choice and outcome remain problematic. METHOD: The present paper reviews the progress in this field since three reviews, all published in the mid-1990s, recommended further attention to three interlinked issues: diagnostic criteria, the symptoms of depression in this group and the lack of rating scales. RESULTS: Despite a further 11 published papers and other studies in progress, the method of diagnosis for people with severe and profound ID remains debatable, with some authors advocating adherence to standard criteria, others suggesting adding criteria to the standard ones and yet others believing that substitute criteria are called for. However, for those with mild to moderate ID, a consensus is emerging that standard diagnostic criteria are appropriate. There has been progress in examining some of the symptoms which might constitute depression in people with ID. New diagnostic criteria issued by the Royal College of Psychiatrists are to be welcomed. There is an assumption in much of the research that symptoms of behaviour commonly termed challenging or maladaptive must be atypical symptoms of depression, but none of the studies reviewed demonstrate this effectively. This is compounded by methodological flaws in the way that depressed samples are arrived at for further study. Although new rating scales have emerged, there is as yet no gold standard diagnostic tool for depression amongst people with ID. CONCLUSIONS: It is suggested that, given these difficulties, the validity of the conceptual frameworks for depression is still in doubt. It remains the case that large-scale, collaborative, prospective studies are called for. PMID- 12558691 TI - Challenging behaviours should not be considered as depressive equivalents in individuals with intellectual disability. AB - BACKGROUND: Depression is one of the most common forms of psychopathology in people with intellectual disability (ID). The present study evaluated the utility of an expanded assessment of psychiatric symptoms and challenging behaviours, as measured by the Clinical Behavior Checklist for Persons with Intellectual Disabilities (CBCPID). METHODS: The CBCPID was administered to 92 people with ID, 35 of whom were diagnosed with a depressive disorder. RESULTS: Item and factor analysis of the scale indicated that depression was best assessed using the core DSM-IV symptoms of depressive disorder. Challenging behaviours such as self injury or aggression were not closely associated with depression. Short scales using the core DSM-IV symptoms of depression were highly internally consistent. There was also evidence of the validity of these scales. CONCLUSIONS: This study found no evidence that challenging behaviours were depressive equivalents in this population. The present authors conclude that the assessment of depression in people with ID should focus on the core DSM-IV symptoms of depression. PMID- 12558692 TI - Glasgow Anxiety Scale for people with an Intellectual Disability (GAS-ID): development and psychometric properties of a new measure for use with people with mild intellectual disability. AB - BACKGROUND: Self-rating scales are widely used in general adult practice; however, there is no reliable and valid method for assessing state anxiety in people with intellectual disability (ID). The present study describes the development and psychometric evaluation of a new scale, the Glasgow Anxiety Scale for People with an Intellectual Disability (GAS-ID). METHODS: First, an item pool was generated from focus groups, a review of the literature and clinician feedback. Secondly, a draft scale was administered to 19 anxious and 16 non anxious people with ID for further validation and appraisal of reliability. Thirdly, the scale was completed by 19 anxious, non-ID people for cross validation with the Beck Anxiety Inventory (BAI). Finally, physiological concomitants were validated by pulse-oximetry. RESULTS: The 27-item GAS-ID discriminated anxious from non-anxious participants, had good test-retest reliability (r = 0.95) and internal consistency (alpha = 0.96), and was reasonably correlated with the BAI (rho = 0.75). The correlation between the physiological subscale of the GAS-ID and changes in pulse rate was moderately significant (rho = 0.52). CONCLUSIONS: This preliminary study suggests that the GAS-ID offers a psychometrically robust and practical (5-10 min) approach to the appraisal of anxiety in this population. PMID- 12558693 TI - Identifying bipolar disorders in individuals with intellectual disability. AB - OBJECTIVE: The aim of the present study was to characterize adults with intellectual disability (ID) and concomitant clinical diagnoses of bipolar disorder (BPD), and determine whether DSM-IV criteria would distinguish individuals with BPD from patients with other psychiatric diagnoses. METHODS: A retrospective chart review was done of a convenience sample of adult patients seen over a 3-year period in a specialty clinic for adults with ID and psychiatric disorders. The DSM-IV criteria were used to differentiate individuals with clinical symptoms of BPD from groups of patients with other mood or thought disorders with behavioural symptoms which frequently overlap those of BPD. Behavioural symptoms were also catalogued and used to distinguish the diagnostic groups. RESULTS: Subjects with clinical symptoms of BPD had significantly more DSM-IV mood-related and non-mood-related symptoms, as well as functional impairments, compared to individuals with major depression, depression with psychosis or schizophrenia/psychosis NOS (not otherwise specified). Likewise, behavioural profiles of the BPD group of patients differed significantly from patients in the other three groups. CONCLUSIONS: Bipolar disorder can be readily recognized and distinguished from other behavioural and psychiatric diagnoses in individuals with ID, and DSM-IV criteria can be useful in the diagnosis of BPD. PMID- 12558694 TI - Patients with and without intellectual disability seeking outpatient psychiatric services: diagnoses and prescribing pattern. AB - BACKGROUND: The present study examined the presenting problem of psychiatric outpatients, and resulting diagnostic and prescribing patterns, comparing patients with intellectual disability (ID) with non-ID (N-ID) patients seen in the same clinic. METHODS: This study was a retrospective medical chart review of information in the first psychiatric diagnostic evaluation for the most recent 100 adult patients with mild ID, 100 patients with moderate, severe or profound ID, and 100 matching N-ID patients. RESULTS: There were significant differences in rates of medical illness, disabilities, history of marriage, children, independent living, and family history of psychiatric and neurological disorders. Individuals with ID were more likely to present with aggression, self-injurious behaviour or physical complaints, whereas N-ID subjects presented more frequently with depression and anxiety complaints. For all groups, depressive disorders were the most frequent class of diagnoses. For those with ID, antipsychotics were used in 32% of subjects, with mood stabilizers in 28% and antidepressants in 27%. The N-ID subjects were most frequently prescribed antidepressants (40%) and anxiolytics (22%). Polypharmacy did not differ significantly among groups. CONCLUSIONS: Psychiatric practitioners relied on the diagnostic examination to formulate their diagnosis, whereas the chief complaint reflected the view of caregivers of the subjects with ID. In contrast to previous studies, outpatient providers frequently diagnosed depression, and the prescribing pattern showed increased usage of antidepressants and mood stabilizers. PMID- 12558695 TI - Prevalence of psychiatric disorders in children and adolescents with and without intellectual disability. AB - BACKGROUND: There have been very few population-based studies of the prevalence of psychiatric disorders among children and adolescents with and without intellectual disability (ID). METHODS: Secondary analysis of the 1999 Office for National Statistics survey of the Mental Health of Children and Adolescents in Great Britain was performed. This survey collected information on a multistage, stratified, random sample of 10 438 children between 5 and 15 years of age across 475 postcode sectors in England, Scotland and Wales. RESULTS: The prevalence of any diagnosed ICD-10 disorder, conduct disorder, anxiety disorder, hyperkinesis and pervasive developmental disorders were significantly greater among children with ID than among their non-ID peers. There were no statistically significant differences between children with and without ID with regard to the prevalence of depressive disorders, eating disorders or psychosis. Factors associated with an increased risk of psychopathology among children and adolescents with ID included age, gender, social deprivation, family composition, number of potentially stressful life events, the mental health of the child's primary carer, family functioning and child management practices. CONCLUSIONS: Children and adolescents with ID are at significantly increased risk of certain forms of psychiatric disorder. Careful consideration of the social and economic adversity facing such families will be necessary to ensure that support services are responsive to both the needs of child as well as the needs of the family in which they are living. PMID- 12558696 TI - Determinants of negative emotional reactions and causal beliefs about self injurious behaviour: an experimental study. AB - BACKGROUND: Caregivers' emotional and cognitive reactions to challenging behaviours have been identified as potential determinants of their reinforcing responses towards such behaviours. However, few studies have explored factors affecting caregivers' emotional and cognitive responses to challenging behaviours. METHODS: Sixty students inexperienced in work with people with challenging behaviours and 60 experienced staff watched one of two carefully matched, acted videotapes depicting self-injury maintained by attention or escape from-task demands. The participants were also told whether the self-injury depicted typically led to mild or severe consequences for the person filmed. The subjects completed measures of their negative emotional reactions to the self injury and their behavioural causal beliefs about the behaviour depicted. RESULTS: Analyses of variance revealed that students reported more negative emotional reactions and were less likely to endorse behavioural causal hypotheses. Those who watched the severe self-injury videotape also reported more negative emotional reactions. Two effects of the behavioural function of the depicted self-injury were also found: (1) attention-maintained self-injury was associated with higher levels of endorsement of behavioural causal hypotheses; and (2) severe attention-maintained self-injury led to the strongest negative emotional reactions, but only from students. CONCLUSIONS: The effects of experience and behavioural function on emotional reactions and behavioural causal beliefs need to be explored in more detail in future research. If replicated, the present results have significant implications for theory and practice in the remediation of challenging behaviours and the support of care staff. PMID- 12558697 TI - From research and development to practice-based evidence: clinical governance initiatives in a service for adults with mild intellectual disability and mental health needs. AB - BACKGROUND: Practice-based evidence represents the contribution of practitioners who utilize research methodologies to examine the quality of their clinical practice and service provision. METHODS: The present paper describes the evolution of a routine practice-based evidence system (PBES) via four phases of research and development. The four phases are described, as is their relevance to assessment and intervention with regard to the mental health problems of people with mild intellectual disability. Phase four describes the development of a routine PBES. RESULTS: The PBES is capable of profiling the individual mental health needs of service users and examining service effectiveness and quality at an organizational level. CONCLUSIONS: The PBES is discussed according to its current utility and possible directions for future development. The system is presented as an example of clinical governance that could be utilized by multidisciplinary teams to develop and maintain an organizational culture of quality. PMID- 12558698 TI - Dietary salt and hypertension: a scientific issue or a matter of faith? AB - Many workers have an overly simplistic view of the relationship between salt intake and hypertension. This article attempts a critical evaluation of some of the evidence. PMID- 12558699 TI - Waiting for scheduled services in Canada: development of priority-setting scoring systems. AB - RATIONALE, AIMS AND OBJECTIVES: An Achilles' heel of Canadian Medicare is long waits for elective services. The Western Canada Waiting List (WCWL) project is a collaboration of 19 partner organizations committed to addressing this issue and influencing the way waiting lists are structured and managed. The focus of the WCWL project has been to develop and refine practical tools for prioritizing patients on scheduled waiting lists. METHODS: Scoring tools for priority setting were developed through extensive clinical input and highly iterative exchange by clinical panels constituted in five clinical areas: cataract surgery; general surgery procedures; hip and knee replacement; magnetic resonance imaging (MRI) scanning, and children's mental health. Several stages of empirical work were conducted to formulate and refine criteria and to assess and improve their reliability and validity. To assess the acceptability and usability of the priority-setting tools and to identify issues pertaining to implementation, key personnel in the seven regional health authorities (RHAs) participated in structured interviews. Public opinion focus groups were conducted in the seven western cities. RESULTS: Point-count scoring systems were constructed in each of the clinical areas. Participating clinicians confirmed that the tools offered face validity and that the scoring systems appeared practical for implementation and use in clinical settings. Reliability was strongest for the general surgery and hip and knee criteria, and weakest for the diagnostic MRI criteria. Public opinion focus groups endorsed wholeheartedly the application of point-count priority measures. Regional health authorities were generally supportive, though cautiously optimistic towards implementation. CONCLUSIONS: While the WCWL project has not 'solved' the problem of waiting lists and times, having a standardized, reliable means of assigning priority for services is an important step towards improved management in Canada and elsewhere. PMID- 12558700 TI - Developing a measure of patient access to primary care: the access response index (AROS). AB - Access to appointments in primary care is not routinely measured, and there is no one standardized method for doing so. Any measurement tool has to take account of the dynamic status of appointment availability and the definitional problems of appointment types. The aim of this study was to develop and trial a method for measuring access that is valid, reliable, quick and provides a daily longitudinal record of access on an organizational basis (not for individual clinicians). Using the results of a literature review and following discussions with clinicians and managers a tool was designed following agreed specifications. After initial adjustments of the tool a feasibility study tested the acceptability of a data collection exercise on 11 practices of varying types, over a 4- to 8-week period. The development phase led to the design of a tool named the access response index (AROS). The method was well received in the practices, with a low incidence of missed days and only one practice failing to return data. The index measures the number of days' wait to the next available appointment with any general practitioner. The inclusion in the score of urgent appointments was abandoned due to definitional problems. A 5-day moving average was chosen to represent the data in graph form to demonstrate overall trends. AROS is a useful tool usable in any practice, and our feasibility study points to it being widely acceptable in the field. Data are represented in clear graphical daily format, either just for one practice or as an anonymous composite graph with other practices in the locality. PMID- 12558701 TI - Can safety assurance procedures in the food industry be used to evaluate a medical screening programme? The application of the Hazard Analysis and Critical Control Point system to an antenatal serum screening programme for Down's syndrome. Stage 1: identifying significant hazards. AB - RATIONALE, AIMS AND OBJECTIVES: Routine review of the Leicestershire district antenatal screening programme for Down's syndrome was carried out by its steering group. This raised suspicions that there were problems with the delivery of the programme. The steering group commissioned a project to adapt the Hazard Analysis and Critical Control Point (HACCP) system, as used in the food industry, to investigate this. The HACCP system is a systematic qualitative approach to problem identification and solution taken from the perspective of the production workers. This paper presents the first stage of the process, the hazard analysis. METHODS: Preliminary work comprised the preparation and verification of a flow diagram of the screening programme. Data about problems (termed hazards) perceived by Leicestershire health care staff delivering the screening programme were collected by observation, interview and telephone. The problems reported were categorized and assessed to identify those of sufficient importance in terms of their consequences or frequency of occurrence to merit the development of control measures. Hazards relating to operational issues were categorized according to the section of the programme to which they related. RESULTS AND CONCLUSIONS: A total of 16 categories of hazard were identified. In addition, nine groups of hazards constituting barriers to delivery of the screening programme as a whole were identified. The HACCP process made these hazards explicit. The first stage of the HACCP system proved a useful tool to identify problems experienced delivering a Down's serum screening programme and to assess which were sufficiently serious to require the development of control measures. PMID- 12558702 TI - Can safety assurance procedures in the food industry be used to evaluate a medical screening programme? The application of the Hazard Analysis and Critical Control Point system to an antenatal serum screening programme for Down's syndrome. Stage 2: overcoming the hazards in programme delivery. AB - RATIONALE, AIMS AND OBJECTIVES: An adaptation of the first stage of the systematic qualitative tool, the Hazard Analysis and Critical Control Point (HACCP) system from the food industry, was used to identify problems in the delivery of a district-wide antenatal screening programme for Down's syndrome. The aim of the second stage, described here, was to develop specific solutions (termed controls) for these problems. METHODS: The views of staff delivering the programme and of members of the Down's screening steering group were used to identify control measures. The steering group members were also asked to identify which controls were critical to a risk management strategy, i.e. whether a control was essential to eliminate the hazard or if other control measures operating further down-line would be sufficient. The control measures were piloted both to determine if they were operationally feasible and whether they were practical in the context of staff workload and attitude. Two main control measures were the development of a rolling education programme for staff and of a written protocol with a checklist to verify stepwise delivery of the Down's screening programme. The checklist was designed to be included in the notes of every pregnant woman to enable audit of the offer of screening and delivery of each step of the screening programme. The control measures were all tailored specifically to overcome the problems reported by staff during the first stage of the work, the hazard analysis. RESULTS AND CONCLUSIONS: The HACCP system proved useful in confirming the existence of problems in delivery of a Down's serum screening programme and in determining solutions tailored to overcome those problems. Future work is required to monitor and evaluate the usefulness of the control measures implemented. PMID- 12558703 TI - Critical incident reporting in UK intensive care units: a postal survey. AB - RATIONALE, AIMS AND OBJECTIVES: To review critical incident reporting in UK intensive care units (ICUs). METHODS: We conducted a postal survey of clinical directors of UK ICUs to ask if critical incident reporting was used in their ICU. Using a structured questionnaire, we also asked for details of critical incident reporting systems and for changes introduced as a result of incident reporting. RESULTS AND CONCLUSIONS: Eighty-six of a total of 256 questionnaires were returned. Sixty-one units had been using a critical incident reporting system for a median duration of 3 years (interquartile range 2-5 years). There were wide variations in the structures of reporting systems between units. A median of four (interquartile range 3-8) critical incidents were reported per unit per month. In 141 changes, the development of protocols and guidelines (18) and changes in drug prescription and checking (20) were most common. A more consistent approach to reporting could improve patient care and 65 responders felt that a national reporting system was potentially useful. PMID- 12558704 TI - Are health states 'timeless'? A case study of an acute condition: post chemotherapy nausea and vomiting. AB - RATIONALE, AIMS AND OBJECTIVES: The objective was to test whether individuals' responses to standard gamble (SG) and visual analogue scale (VAS) questions do not depend on the time horizon of the health scenario presented. METHODS: Face-to face interviews were conducted in a convenience sample of 18 women aged 22-50 years with no history of breast cancer or cancer requiring chemotherapy. Data were collected from March 2000 to June 2000 at a university in the Midwest of the United States of America. Preference weights were estimated using SG top-down titration method and VAS scaled from zero (death) to one (perfect health). Subjects were asked to rate their preferences if faced with two scenarios: post chemotherapy nausea and vomiting (PCNV) occurring for 3 days (scenario 1), and PCNV lasting for the rest of their lives (scenario 2). Three PCNV health states of varying severity were tested: complete alleviation, partial alleviation, and no alleviation. RESULTS: Paired-t-test analysis showed statistically significantly lower preference weights (P < 0.05) when the health state was for the rest of the respondent's life vs. 3 days. Mean SG weights for scenario 1 vs. scenario 2 were: 0.968 vs. 0.927 (complete alleviation), 0.942 vs. 0.810 (partial alleviation) and 0.866 vs. 0.644 (no alleviation). Mean VAS weights for scenario 1 vs. scenario 2 were: 0.741 vs. 0.676 (complete alleviation), 0.490 vs. 0.307 (partial alleviation) and 0.276 vs. 0.136 (no alleviation). DISCUSSION AND CONCLUSIONS: For the majority of respondents the utility independence assumption for SG and VAS did not hold. Similar to Bala et al., the results of this study indicated that preference weights as measured by SG and VAS techniques were not 'timeless'. Regardless of the preference measure used, both SG and VAS yielded higher scores when PCNV lasted for a shorter period of time. PMID- 12558705 TI - Evaluation of chloroquine as a potent anti-malarial drug: issues of public health policy and healthcare delivery in post-war Liberia. AB - Chloroquine-resistant plasmodium falciparum malaria is a serious public health threat that is spreading rapidly across Sub-Saharan Africa. It affects over three quarters (80%) of malarial endemic countries. Of the estimated 300-500 million cases of malaria reported annually, the vast majority of malarial-related morbidities occur among young children in Africa, especially those concentrated in the remote rural areas with inadequate access to appropriate health care services. In Liberia, in vivo studies conducted between 1993 and 2000 observed varying degrees of plasmodium falciparum malaria infections that were resistant to chloroquine, including sulfadiazine-pyrimethamine. As the country emerges from a prolonged civil war, the health care delivery system may not be adequately prepared to implement an effective nation-wide malarial control strategy. As a result, the management of uncomplicated malaria in Liberia poses a significant public health challenge for the government-financed health care delivery system. Therefore, based on extensive literature review, we report the failure of chloroquine as an effective first-line drug for the treatment of uncomplicated plasmodium falciparum malaria in Liberia and recommend that national health efforts be directed at identifying alternative drug(s) to replace it. PMID- 12558706 TI - Health vs. disease: a commentary on 'The rationale of value-laden medicine' (Kottow 2002; Journal of Evaluation in Clinical Practice 8, 77-84). PMID- 12558707 TI - Critical thinking vs. moral expertise: a commentary on 'The rationale of value laden medicine' (Kottow 2002; Journal of Evaluation in Clinical Practice 8, 77 84). PMID- 12558709 TI - Healing pressure ulcers with collagen or hydrocolloid: a randomized, controlled trial. AB - OBJECTIVES: To compare the effects of topical collagen and hydrocolloid on pressure ulcer healing. DESIGN: Randomized (allocation concealed), single-blind (outcome assessors), controlled trial with 8-week follow-up. SETTING: Eleven nursing homes in central Illinois. PARTICIPANTS: Sixty-five patient-residents with Stage II or III pressure ulcers: median age 83.1, median Braden score 12, 63% female, 80% Stage II ulcers, and 20% Stage III ulcers. Exclusion criteria included cellulitis and osteomyelitis. INTERVENTION: Thirty-five patients were allocated to topical collagen daily, 30 to topical hydrocolloid twice weekly. MEASUREMENTS: The primary outcome was complete healing within 8 weeks. Secondary outcomes were time to heal, ulcer area healed per day, linear healing of wound edge, and cost of therapy. RESULTS: Analysis by intention to treat revealed similar complete ulcer healing within 8 weeks in collagen (51%) and hydrocolloid (50%) recipients (difference 1%, 95% confidence interval (CI) = 26-29%). Mean healing time was similar: collagen healed in 5 weeks (95% CI = 4-6), hydrocolloid healed in 6 weeks (95% CI = 5-7). Mean area healed per day was 6 mm(2)/d in both treatment groups. Mean linear healing of the wound edge was 3 mm in both groups. In multivariate analysis, baseline ulcer depth was the only independent predictor of complete ulcer healing within 8 weeks (odds ratio = 0.56, 95% CI = 0.38-0.81). Cost analysis favored hydrocolloid. CONCLUSIONS: There were no significant differences in healing outcome between collagen and hydrocolloid. Collagen was more expensive and offered no major benefits to patients otherwise eligible for hydrocolloid treatment. PMID- 12558710 TI - Comparison of subcutaneous and intravenous rehydration in geriatric patients: a randomized trial. AB - OBJECTIVES: To compare the acceptance, feasibility, and adverse effects of subcutaneous (SC) and intravenous (IV) rehydration in dehydrated geriatric patients and clinical changes exhibited by the patients. DESIGN: A prospective, randomized, open clinical trial. SETTING: Hospital geriatric wards during a period of 20 months. PARTICIPANTS: Ninety-six patients with a mean age +/- standard deviation of 85.3 +/- 6.7 with signs of mild to moderate dehydration needing parenteral fluids. INTERVENTION: Geriatric patients were randomly allocated to receive SC or IV infusions of half-normal saline-glucose solutions as long as clinically necessary. MEASUREMENTS: A standardized patient record form was used to document the observed adverse effects. Using a Likert-like scale based on the German school marks system with scores ranging from 1 = very good to 6 = very bad, patients were asked to score their discomfort; nurses and doctors scored the feasibility of the intervention. Changes in laboratory and clinical findings (including patients' orientation and activities of daily living using the Barthel Index) and adverse effects were recorded. RESULTS: Forty-eight patients were randomized into each group. Median duration of fluid administration was 6 days (SC and IV, P =.33). Median volume was 750 mL/day (SC) and 1,000 mL/day (IV, P =.002). In 13 patients, the therapy had to be changed from SC to IV (SC/IV subgroup): 11 times because of the exigency of an IV drug application and twice because of poor resorption. In 17 patients, there was a change from IV to SC (IV/SC subgroup), mainly because of impossibility of further peripheral IV punctures (8 times) and permanent removal of the IV cannula (5 times). The patients of the IV/SC subgroup scored their discomfort significantly worse (median 5.5 vs all other groups median 2, P =.017). This corresponded with the scoring of feasibility by the nurses (IV/SC: median 4.25 vs all other groups median 2, P =.009) and by the doctors (IV/SC: median 4 vs all other groups: median 2, P =.001). Both methods of rehydration caused only few systemic adverse reactions; acute cardiac failure occurred twice in the SC group versus four times in the IV group (P =.68) and hyponatremia once in the SC group versus twice in the IV group (P = 1.0). Some patients experienced local side effects (SC, n = 29 vs IV, n = 24; P =.41), mainly to a mild extent (SC, n = 25 vs IV, n = 24; P = 1.0). Major local side effects (large edema, phlebitis, cellulitis, erythema and strong pain) occurred in nine SC and eight IV (P = 1.0) patients. The clinical and laboratory changes during therapy were similar in both trial arms. CONCLUSIONS: Rehydration by hypodermoclysis is equally well accepted by geriatric patients as the IV therapy and offers a similarly easy feasibility. Additionally, in confused patients and in those in whom IV punctures are difficult to achieve, it represents the far superior method. Both techniques are comparably safe and effective. PMID- 12558711 TI - Does an exercise and incontinence intervention save healthcare costs in a nursing home population? AB - OBJECTIVES: To determine whether an intervention that combines low-intensity exercise and incontinence care offsets some of its costs by reducing the incidence of selected health conditions in nursing home residents. DESIGN: Randomized, controlled trial with the incidence and costs of selected, acute conditions compared between a 6-month baseline and an 8-month intervention phase. SETTING: Four nursing homes. PARTICIPANTS: One hundred ninety incontinent, long stay nursing home residents. INTERVENTION: Low-intensity, functionally oriented exercise and incontinence care were provided every 2 hours from 8:00 a.m. to 4:00 p.m. for 5 days a week for 8 months. MEASUREMENTS: Predefined acute conditions hypothesized to be related to physical inactivity, incontinence, or immobility were abstracted from residents' medical records by blinded observers during a 6 month baseline period and throughout the 8-month intervention. Conditions included those in the dermatological, genitourinary, gastrointestinal, respiratory and cardiovascular systems; falls; pain; and psychiatric and nutritional disturbances. Costs were determined using Current Procedural Terminology Center and Medicare allowable cost reimbursement at a rate of 80%. RESULTS: The intervention group had significantly better functional outcomes than the control group (strength, mobility endurance, urinary and fecal incontinence) and a reduction of 10% in the incidence of the acute conditions, which was not significant. There were no significant differences between groups in the cost of assessing and treating these acute conditions between baseline and intervention. CONCLUSION: The intervention, which is consistent with federal and clinical practice guidelines, significantly improved functional outcomes but did not reduce the incidence and costs of selected acute health conditions. The cost of implementing these labor-intensive interventions for frail nursing home residents will have to be justified based on functional and quality-of-life outcomes and are unlikely to be offset by savings in medical care costs in this population. PMID- 12558712 TI - Prevalence of dementia in older latinos: the influence of type 2 diabetes mellitus, stroke and genetic factors. AB - OBJECTIVES: To estimate dementia prevalence in older Mexican Americans, determine the distribution of dementia by etiology, and evaluate the contribution of type 2 diabetes mellitus, stroke, and apolipoprotein E (ApoE) genotype to dementia. DESIGN: Analysis of baseline data from an epidemiological cohort study. SETTING: Sacramento Valley, California. PARTICIPANTS: One thousand seven hundred eighty nine Latinos aged 60 and older residing in targeted census tracts during 1998-99. MEASUREMENTS: Each subject was interviewed and screened for dementia and cardiovascular risk factors and diseases. Fasting blood samples were drawn for glucose, insulin, and lipids. Buccal cells were obtained for genetic analysis of ApoE. A three-stage process of screening was used to diagnose dementia, including cognitive testing, a clinical examination, and imaging to determine etiology. Presence of dementia was established according to National Institute of Neurological Disorders and Stroke/Alzheimers and Related Disorders Association criteria and California Alzheimer's Disease Diagnostic and Treatment Criteria. RESULTS: Overall dementia prevalence was 4.8%. Prevalence in those aged 85 and older was 31%. Education and Anglo cultural orientation was negatively associated with dementia risk. Risk of dementia was nearly eight times higher in those with both type 2 diabetes mellitus and stroke. Forty-three percent of dementia was attributable to type 2 diabetes mellitus, stroke, or a combination of the two. ApoE allele frequency was E2 5.9%, E3 90.1%, and E4 4%. Those with any E4 and 4-4 combinations had a higher risk for dementia than those with the E3-3 combination. CONCLUSIONS: Dementia prevalence in this ethnic group is similar to that reported in Canadian and European studies but lower than in Caribbean-Hispanics residing in the United States. The etiological fraction of dementia attributable to type 2 diabetes mellitus and stroke is substantial and points toward the need for intervention research and treatment with the goal of reducing neurological sequelae in groups with high prevalence of type 2 diabetes mellitus. The allele frequency of ApoE was similar to that in other published studies on Mexican Americans. The low frequency of the E4 allele may contribute to the difference in etiology of dementia in older Mexican Americans and older people of European background. Dementia in this ethnic group may be related to preventable causes, with a smaller genetic component than in Europeans. PMID- 12558713 TI - Cognitive impairment and mortality in older mexican americans. AB - OBJECTIVES: To examine the extent to which cognitive status and decline in cognitive status predict mortality in older Mexican Americans. DESIGN: Longitudinal cohort. SETTING: Older Mexican Americans residing in five southwestern states in the United States. PARTICIPANTS: Two thousand six hundred twenty-five persons aged 65 and older living in Texas, New Mexico, Colorado, Arizona, and California. MEASUREMENTS: The cognitive function of participants was assessed using the Mini-Mental State Examination (MMSE) in 1993-94 and again in 1995-96. Cognitive decline was defined using two sets of criteria: a drop to 17 or less (moderate-severe cognitive impairment) on the MMSE at 2-year follow-up and a decrease of at least four points, a threshold change in scores that was used to predict mortality in this sample. Cox proportional hazards models were conducted to examine the association between the MMSE and increased risk for mortality, after controlling for sociodemographic characteristics, medical conditions, and depressive symptoms. RESULTS: Five-year mortality risk was significantly associated with persons in the moderately-severely cognitively impaired category (hazard ratio (HR) = 2.35, P <.001). Moreover, mild cognitive impairment was also predictive of mortality in older Mexicans (HR = 1.45, P <.001). Two-year declines in cognitive functioning, particularly in persons who had declined to the moderate-severe impairment category (HR = 2.23, P <.001) and those who dropped at least four points on the MMSE (HR = 1.30, P <.001), were predictive of mortality 3 years later, after controlling for important variables. CONCLUSIONS: Baseline moderate-severe and mild cognitive status and 2-year decline in cognitive functioning independently predict mortality in older Mexican Americans. Although age and selected medical conditions have been reported as the more salient predictors of mortality, cognitive functioning should be considered part of identifying older persons at high risk for underlying medical conditions and mortality. PMID- 12558715 TI - The incremental effect of dementia-related problem behaviors on the time to nursing home placement in poor, frail, demented older people. AB - OBJECTIVES: To examine the incremental effect of dementia-related problem behaviors (DRPBs) on the risk of and time to nursing home placement (NHP) in poor, frail, demented older people. DESIGN: Client assessments were reviewed retrospectively for clinical, functional, and cognitive information, particularly the presence of DRPBs and the time during enrollment at which they occurred. SETTING: The Georgia Community Care Services Program (CCSP), a Medicaid 1915-C home and community-based services program (HCBS). PARTICIPANTS: A random sample of demented clients (n = 204) discharged during fiscal year 1996 from four CCSP regions. MEASUREMENTS: The risk of and time to nursing home placement. Cox proportional hazards models and an extended Cox model with a time-varying covariate for the presence of DRPBs were used to evaluate the risk factors associated with NHP in demented older people. Kaplan-Meier survival curves were used to estimate the time to NHP for several risk groups. RESULTS: In demented older people in a HCBS program, having a DRPB was found to increase the hazard rate of NHP. Having a problem behavior shortened average median survival in the community by approximately 2 years. CONCLUSIONS: DRPBs in demented older people shorten the time to NHP. Exploration of effective interventions is warranted. PMID- 12558714 TI - Survival to late dementia in Dutch nursing home patients. AB - OBJECTIVES: To investigate the determinants of survival to late dementia in Dutch nursing home patients. DESIGN: Observational analysis of a cohort of patients with a prospective follow-up. SETTING: Psychogeriatric nursing home "Joachim en Anna" in Nijmegen, the Netherlands. PARTICIPANTS: Dementia patients from a psychogeriatric nursing home admitted between 1980 and 1989. MEASUREMENTS: All patients were followed until death. Age, sex, severity of dementia on admission, and type of dementia were analyzed as determinants of survival to late dementia. Late dementia was operationally defined as total impairment on 20 items of the functional status questionnaire. RESULTS: The mean disease duration of the study population was 7.0 years, with a wide range. Some died soon after dementia was diagnosed, whereas others lived for more than 25 years after onset of the disease. One hundred twenty-six patients (14.2%) reached the phase of late dementia, most of whom were women (90%) with Alzheimer's disease (60%). For most of the patients, this phase lasted 1 year or less. Age, sex, type of dementia, and severity of dementia on admission all contributed significantly to predicting survival to late dementia. CONCLUSION: One of seven nursing home patients with dementia survived to late dementia. Nursing home physicians can use these findings to inform relatives about the prognosis. PMID- 12558716 TI - The yield, reliability, and validity of a postal survey for screening community dwelling older people. AB - OBJECTIVES: To assess the yield, reliability, and validity of a postal survey developed to identify older persons in need of outpatient geriatric assessment and follow-up services. DESIGN: A longitudinal cohort study. SETTING: Outpatient primary care clinic at a Department of Veterans Affairs teaching ambulatory care center. PARTICIPANTS: Patients (N = 2,382) aged 65 and older who returned a Geriatric Postal Screening Survey (GPSS) that screened for common geriatric conditions (depression, cognitive impairment, urinary incontinence, falls, and functional status impairment). Validity and reliability testing was performed with subsamples of patients classified as high or lower risk based on responses to the GPSS. MEASUREMENTS: Test-retest reliability was measured by percentage agreement and kappa statistic. The diagnostic validity of the 10-item GPSS was tested by comparing single GPSS items to standardized geriatric assessment instruments for depression, mental status and functional status, as well as direct questions regarding falls, urinary incontinence, and use of medications. Validity was also tested against clinician evaluation of the specific geriatric conditions. Predictive validity was tested by comparing GPSS score with 1-year follow-up data on functional status, survival, and healthcare use. RESULTS: Respondents identified as high risk by the GPSS had scores that indicated significantly greater impairment on structured assessment instruments than those identified as lower risk by GPSS. The overall mean percentage agreement between the test and retest surveys was 88.3%, with a mean weighted kappa of 0.70. In comparison with a structured telephone interview and with a clinical assessment, individual items of the GPSS showed good accuracy (range 0.71-0.78) for identifying symptoms of depression, falls, and urinary incontinence. Over a 1 year follow-up period, the GPSS-identified high-risk group had significantly (P <.05) more hospital admissions, hospital days and nursing home admissions than the lower-risk group. CONCLUSION: A brief postal screening survey can successfully target patients for geriatric assessment services. In screening for symptoms of common geriatric conditions, the GPSS identified a subgroup of older outpatients with multiple geriatric syndromes who were at increased risk for hospital use and nursing home admission and who could potentially benefit from geriatric intervention. PMID- 12558717 TI - Comparison of mammography use by older black and white women. AB - OBJECTIVES: To identify differences in the prevalence of ever having had a mammogram and having had a recent mammogram between older black and white women and to compare factors associated with mammography use in older black and white women. DESIGN: Data analysis and comparative study using nationally representative multistage sampling survey. SETTING: Data were obtained from the 1998 National Health Interview Survey. PARTICIPANTS: Four hundred forty-nine black and 3,328 white older women were examined. MEASUREMENTS: The outcome variables included never having had a mammogram (yes/no) and not having had a mammogram in the past 3 years (yes/no). RESULTS: The results of chi-square tests showed that older blacks were less likely to have ever had a mammogram than older whites, but there was no difference in having had a recent mammogram between older blacks and whites. After adjusting for other related factors, race was not related to mammography use in older blacks and whites. Health insurance was related to mammography use in older whites but not in older blacks. Family income was associated with never having had a mammogram in older whites but not in older blacks. Older blacks with less than 12 years of education were less likely to have had a mammogram (recently or ever) than older whites with less than 12 years of education. CONCLUSIONS: Even though race, per se, was not associated with mammography use in older black and white women, many barriers to mammography use between older black and white women were different or did not have similar effects. To promote mammography use in older black and white women, barriers need to be specifically targeted for each group to enhance the effectiveness of breast cancer screening programs. PMID- 12558718 TI - Mortality-related factors and 1-year survival in nursing home residents. AB - OBJECTIVES: To identify factors associated with 1-year mortality in newly admitted and long-stay (in nursing home longer than 1 year) nursing home residents by linking Minimum Data Set (MDS) information with data from the National Death Index and use these factors to create a useful tool for estimating risk levels for 1-year mortality. DESIGN: Retrospective cohort study with development and validation cohorts. SETTING: All 643 Medicare and Medicaid certified nursing homes in New York State during the study period. PARTICIPANTS: The study included data on residents collected during full MDS assessments from June 1994 through December 1997. A total of 100,669 nursing home residents met the inclusion criteria for the newly admitted resident analysis. The newly admitted development cohort included 60,341 residents, and the newly admitted validation cohort included 40,328 residents. A total of 36,125 nursing home residents met inclusion criteria for the long-stay (residing in nursing home>1 year) cohort. The long-stay development cohort included 22,749 residents, and the long-stay validation cohort included 15,068 residents. MEASUREMENTS: The analytical approach was similar for the newly admitted and long-stay resident cohorts. Resident characteristics that were considered potential risk factors for mortality were examined individually in bivariate proportional hazards models, and factors with P <.05 were entered into a proportional hazards regression stepwise model. The strongest factors based on their chi-square values were selected for entry into a multivariate proportional hazards analysis. Hazard ratios (HRs), 95% confidence intervals, and P-values were derived from this model. A mortality risk index score was created for each resident by summing the value of each HR in the multivariate model for those who had the risk factor. A sensitivity analysis was performed to determine the effect of residents with an unknown death status. A similar analysis was performed on the validation cohort to validate the original results. RESULTS: Major factors associated with 1-year mortality were identified in both the newly admitted and long-stay cohorts. In both newly admitted and long-stay residents, a higher mortality risk index score was associated with increased 1-year mortality in both the development and validation cohorts. CONCLUSIONS: MDS data can identify major factors associated with 1-year mortality in newly admitted and long-stay nursing home residents. These factors can be used to stratify residents into risk categories for 1-year mortality. This information could be important to residents, their families, and their physicians when developing care plans, as well as to agencies interested in healthcare resource planning. PMID- 12558719 TI - Sex differences in peripheral arterial disease: leg symptoms and physical functioning. AB - OBJECTIVES: To compare lower extremity functioning and leg symptoms between women and men with peripheral arterial disease (PAD). DESIGN: Cross-sectional. SETTING: Three Chicago-area medical centers. PARTICIPANTS: One hundred eighty-seven women and 273 men with PAD identified consecutively in patients in the noninvasive vascular laboratories and a general medicine practice at the three medical centers. MEASUREMENTS: Walking speed, 6-minute walk, accelerometer-measured 7-day physical activity, and a summary performance score. The summary performance score combines data on walking velocity, time for five repeated chair rises, and standing balance to achieve a score on a 0 to 12 scale (12 = best). RESULTS: Women with PAD were older and had a lower prevalence of prior leg revascularization, a higher prevalence of spinal stenosis, and a lower prevalence of other cardiovascular disease than men with PAD. Mean ankle brachial index (ABI) values +/- standard deviation were similar in women and men with PAD (0.64 +/- 0.15 vs 0.66 +/- 0.14, P =.15). Women with PAD were significantly more likely than men with PAD to have exertional leg pain that sometimes begins at rest (27.8% vs 13.2%, P <.001). Women with PAD had slower walking speed (0.81 vs 0.92 m/s, P <.001), shorter 6-minute walk distance (1,047 vs 1,182 feet, P <.001), and a poorer summary performance score (8.9 vs 9.8, P <.001) than men with PAD, adjusting for age, race, height, comorbid disease, and leg symptoms. After adjusting for leg strength, sex differences in 6-minute walk performance and summary performance score were attenuated modestly (1,089 vs 1,177 feet for 6 minute walk, P =.022 and 9.2 vs 9.8 for summary performance score, P =.027). CONCLUSION: Women with PAD had a higher prevalence of leg pain on exertion and rest, poorer functioning, and greater walking impairment from leg symptoms than men with PAD. A higher prevalence of spinal stenosis in women may explain the observed sex differences in leg symptoms. Poorer leg strength in women may contribute to poorer lower extremity functioning in women with PAD than in men with PAD. PMID- 12558720 TI - Effect of end-stage renal disease on the quality of life of older patients. AB - OBJECTIVES: To assess the effect of chronic renal failure (CRF) on quality of life (QOL). DESIGN: Controlled cross-sectional study. SETTING: All 13 dialysis units in the French Lorraine region and six departments of a Nancy hospital. PARTICIPANTS: One hundred sixty-nine older patients with end-stage renal disease (ESRD) who were starting first dialysis (mean age +/- standard deviation = 76.2 +/- 5.1) and 169 age- and sex-matched non-CRF controls. MEASUREMENTS: QOL was assessed using the short-form health survey 36-item (SF-36) self-administered questionnaire. Information on comorbid conditions, clinical symptoms and laboratory findings was obtained from medical records. RESULTS: Mean QOL scores ranged from 11.2 (role limitation due to physical function (RP) dimension) to 55.5 (social function) in the ESRD group, and from 22.0 (RP) to 54.3 (mental health) in the non-CRF group. Among ESRD patients, factors related to QOL scores (physical function (PF) and vitality dimensions) were the conditions under which dialysis was initiated and comorbidity. Patients whose first dialysis was unplanned had 10.4 fewer points in the PF dimension than those in whom it was planned (P =.014). CONCLUSIONS: If dialysis initiation is planned, ESRD in older patients has no more effect on QOL than others diseases. However, patients whose dialysis is unplanned have severely impaired QOL. These results represent an argument for improving the predialysis care of older renal failure patients to optimize conditions at first dialysis. PMID- 12558721 TI - Physicians vary in approaches to the clinical management of delirium. AB - OBJECTIVES: To ascertain the variation in strategies for managing delirium of physicians with expertise in geriatrics. DESIGN: Cross-sectional mail survey. SETTING: United States. PARTICIPANTS: A probability sample of physician members of the American Geriatrics Society. MEASUREMENTS: Management choices presented in a two-part case vignette of an older woman hospitalized with a hip fracture who develops mild and then severe delirium. RESULTS: One hundred twenty-two respondents (43%) selected the three answers constituting current "best practice," 50 (18%) selected an unnecessary diagnostic test (brain imaging, lumbar puncture, or electroencephalogram), and 47 (17%) selected unnecessary pharmacologic therapy for mild delirium. For severe delirium, 270 (96%) selected pharmacological therapy, of whom 180 chose haloperidol alone, 55 chose lorazepam alone, 23 chose lorazepam in combination with haloperidol, and 12 wrote in another drug. Thirty percent of the respondents made any selection of lorazepam, alone or in combination with haloperidol, for mild or severe delirium. Sixty-one percent of those selecting haloperidol for severe delirium chose a dose greater than that recommended for geriatric patients. Sex, date of graduation from medical school, clinical specialty, completion of a geriatric fellowship, or certification in geriatrics had no significant effect on responses. CONCLUSIONS: The common selection of lorazepam to treat delirium is troubling because benzodiazepines themselves are implicated in delirium. Selection of an initial dose of haloperidol higher than that recommended for geriatric patients by more than half of the respondents is also of concern. There is a paucity of sound clinical evidence to guide the choice of pharmacological agents for treating delirium in older hospitalized patients. PMID- 12558722 TI - Pneumococcal polysaccharide revaccination: immunoglobulin g seroconversion, persistence, and safety in frail, chronically ill older subjects. AB - OBJECTIVES: To determine the 1-month postpneumococcal polysaccharide revaccination immunoglobulin G (IgG) antibody response, its persistence at 1 year, and tolerability of revaccination in frail, chronically ill older nursing facility residents. DESIGN: Prospective study conducted between December 1998 and July 2000. SETTING: Six skilled nursing facilities in the Minneapolis-St. Paul, Minnesota, metropolitan area. PARTICIPANTS: Sixty-seven subjects aged 65 and older having received primary vaccination with pneumococcal polysaccharide vaccine (PPV) at least 5 years before enrollment. INTERVENTION: Revaccination with one dose of 23-valent PPV. MEASUREMENTS: Adverse events and concentrations of seven individual pneumococcal polysaccharide type-specific IgG antibodies (against serotypes 4, 6B, 9V, 14, 18C, 19F, 23F) and their aggregate before and 1 and 12 months after revaccination. RESULTS: A significant increase in all individual and aggregate median antibody concentrations over baseline was observed 1 month after revaccination. However, after 1 year, the increase remained significant only for serotypes 6B and 18C and the aggregate parameter. One month after revaccination, the mean increase in antibody concentration over baseline was significantly greater than 1.4-fold for six of the seven serotypes and the aggregate. However, the increase was not significantly greater than 1.4 at 1 year for any of the serotypes or the aggregate. Minor, self-limited localized adverse reactions and systemic reactions occurred in 11.3% of the subjects. CONCLUSIONS: In frail, chronically ill older nursing facility residents, revaccination with 23-valent PPV at least 5 years after primary vaccination (whether primary vaccination occurred before or after age 65) is associated with a significant, albeit brief, immunological response for most of the serotypes tested. Revaccination was well tolerated. PMID- 12558723 TI - Differences in mortality of black and white patients enrolled in the program of all-inclusive care for the elderly. AB - OBJECTIVES: To examine the relationship between race and mortality in frail community-dwelling older people with access to a program providing comprehensive access and coordination of services. DESIGN: A longitudinal cohort study. SETTING: Twelve nationwide demonstration sites of the Program of All-Inclusive Care for the Elderly (PACE) from 1990 to 1996. PACE provides comprehensive medical and long-term care services for nursing home-eligible older people who live in the community. PARTICIPANTS: Two thousand two white patients and 859 black patients. MEASUREMENTS: Patients were followed after enrollment until death or the end of the follow-up period. Time from enrollment to death was measured with adjustment of the Cox proportional hazards model for comorbid conditions, functional status, site, and other demographic characteristics. RESULTS: Black patients were younger than white patients (mean age 77 vs 80, P <.001) but had worse functional status (mean activity of daily living (ADL) score 6.5 vs 7.2, P <.001) on enrollment. Survival for black and white patients was 88% and 86% at 1 year, 67% and 61% at 3 years, and 51% and 42% at 5 years, respectively (unadjusted hazard ratio (HR) for black patients = 0.77; 95% confidence interval (CI) = 0.67-0.89). After adjustment for baseline comorbid conditions, functional status, site, and demographic characteristics, black patients still had a lower mortality rate (HR = 0.77; 95% CI =.65-0.93). The survival advantage for black patients did not emerge until about 1 year after PACE enrollment (HR for first year after enrollment = 0.97; 95% CI = 0.72-1.31; HR after first year = 0.67; 95% CI = 0.54-0.85, P-value for time interaction <.001). During the first year of enrollment, black patients were more likely to improve and less likely to decline in ADL function than white patients (P <.001). CONCLUSION: In PACE, a system providing access to and coordination of comprehensive medical and long-term care services for frail older people, black patients have a lower mortality rate than white patients. This survival advantage, which emerges approximately 1 year after PACE enrollment, may be related to the comprehensive access and coordination of services provided by the PACE program. PMID- 12558724 TI - Development and validation of an instrument to predict probability of home discharge from a geriatric convalescence unit in Spain. AB - OBJECTIVES: To develop and validate an instrument to predict probability of home discharge upon hospital admission. DESIGN: Prospective cohort study. SETTING: Geriatric convalescence unit (GCU) in Spain. PARTICIPANTS: Seven hundred eighty one patients aged 65 and older consecutively admitted to a GCU over a 4-year period. The total sample was randomized and divided into two subgroups; the first (n = 575) was used to construct the predictive instrument (development subgroup) and the second (n = 206) for the validation process (validation subgroup). MEASUREMENTS: All patients were evaluated within the first 72 hours after admission. Age, sex, functional status before admission, diagnostic categories, functional status on admission, comorbidity, cognitive function, and social support were assessed. RESULTS: Logistic regression analysis identified three patient characteristics as independent predictors of home discharge in the development subgroup: higher scores on functional status at admission (Barthel index), normal Mini-Mental State Examination scores, and lower scores on Social Familial Evaluation Scale. A scoring system ranging from 0 to 5 was constructed using these variables to predict probability of home discharge (PHD). Different PHD scores (0-1, 2, 3, 4, and 5) identified patients with different probabilities of returning home on discharge in the validation subgroup (36.5%, 53.6%, 60.8%, 83.3%, and 100%, respectively). PHD scores of 4 and 5 demonstrated substantially higher posttest than pretest probability, with moderate and high clinical effect value. Scores of 0 or 1 demonstrated substantially lower posttest than pretest probability. CONCLUSION: A PHD instrument may be useful in identifying patients most likely to be discharged to home from the GCU. Patients with low probability of home discharge may also be identified early. PMID- 12558725 TI - The value of positron emission tomography in the clinical evaluation of dementia. AB - Positron emission tomography (PET) has been promoted as a means of improving the diagnosis of Alzheimer's disease (AD), but the evidence to support its incremental value is unclear. To assess the evidence regarding the use of PET in the clinical evaluation of AD, a systematic review of the English-language literature indexed in MEDLINE (1975-January 2001), the Cochrane Library (issue 4, 2000), and health technology assessment (HTA) reports was conducted. Articles identified by this review process were graded for methodological and reporting quality using a standardized grading scheme. Sixteen original articles and seven HTA reports were identified. In general, the articles addressed: using PET to differentiate AD from normal aging or non-Alzheimer's dementias, PET imaging compared with single positron emission computed tomography imaging, using PET to predict the progression of dementia, and agreement and reliability in the interpretation of PET images. Serious problems with study design and methodology in all articles were identified. Previous HTA reports have generally recommended that PET not be used in the clinical evaluation of dementia. In conclusion, there is little evidence to support the addition of PET to the routine clinical evaluation of patients with suspected or established dementia. Suggestions for future research in this area are offered. PMID- 12558726 TI - Preventing disability through community-based health coaching. AB - The California Public Employees Retirement System (CalPERS) Health Matters program is a randomized controlled trial of a community-based health coaching program operating in Sacramento, California, since January 2001. It is modeled after the Health Enhancement Program and Senior Wellness Program of Seattle, Washington. Like the Health Enhancement Program, this program incorporates a menu of disability-prevention strategies, with health coaching, patient education on the self-management of chronic illness, and fitness forming the program's core. Unlike the Health Enhancement Program, the Health Matters program focuses as much attention on linking participants to existing community, health plan, and self directed programming as it does encouraging them to participate in programming developed especially for the project. All participants and controls continue to receive their usual medical care from their managed care providers. Eligibility criteria for the program include having one or more qualifying chronic health conditions, being aged 65 and older, being a member of a participating health plan, and being accepted into CalPERS' Long Term Care Insurance Program. Baseline exclusions include being cognitively impaired or qualifying for long-term care insurance benefits due to deficiencies in two or more activities of daily living. The project has been successful in its enrollment strategy. It has also been successful in recruiting participants into project-sponsored, community-based, and self-directed disability-prevention programming. This is particularly true for programming related to diet, exercise, and various aspects of disease management. PMID- 12558727 TI - Health screening decisions for older adults: AGS position paper. PMID- 12558728 TI - Treatment of diabetes mellitus in older people: oral therapy options. PMID- 12558729 TI - Evaluating an incontinence intervention in nursing home residents. PMID- 12558730 TI - The "Alzheimerization" of dementia research. PMID- 12558731 TI - Vitamin B12 and methyl malonic acid: do we have all the answers? PMID- 12558732 TI - Sertraline and vaginal bleeding--a possible association? PMID- 12558733 TI - Frontotemporal dementia associated with vitamin B12 deficiency. PMID- 12558734 TI - Care of patients near death: another view. PMID- 12558735 TI - Rehabilitation outcomes in frail older patients. PMID- 12558736 TI - A case report: doctor, my daughter has Alzheimer's disease. PMID- 12558737 TI - Demonstration of a foamy plug in the main bronchus using chest computed tomography. PMID- 12558738 TI - Resting heart rate in older people: a predictor of survival to age 85. PMID- 12558740 TI - Policy on ethics and patient consent. PMID- 12558741 TI - Legal and ethical issues in the use of anonymous images in pathology teaching and research. AB - The privacy of patients' health information is of paramount importance. However, it is equally important that medical staff and students have access to photographs and video recordings of real patients for training purposes. Where the patient can be identified from such images, his or her consent is clearly required to both obtain the image and to use it in this way. However, the need for consent, both legally and ethically, is much less convincing where the patient cannot, by the very nature of the image, be identified from it. This is the case for many images used in the teaching of clinical medicine, such as videos taken of laparoscopies, images of internal organs and unlabelled X-rays. PMID- 12558742 TI - The development and evaluation of the UK national telepathology network. AB - AIMS: As technology advances and costs fall, it may be anticipated that soon every histopathologist will expect to be able to exchange electronic images with colleagues. Arguing that the value of a network increases as more people are connected, we sought to install a simple, low-cost telepathology system into any histopathology laboratory which requested it within the UK. METHODS AND RESULTS: We assumed that laboratories had microscopes, computers and internet access. We offered low-cost video cameras, video input cards, software and training to any histopathology department requesting installation, limited only by resources supplied by the UK government. We also established central servers and a website with 'help' files. After 1 year we studied system use and pathologists' opinions by circulating a questionnaire. Installations were completed in 35 laboratories; there are currently 66 registered users of the system, with 16 identified 'experts' covering most organ systems. Serious difficulties were caused by institutional firewalls and reluctance of local information technology (IT) staff to make changes to facilitate the installation or to help resolve subsequent network problems. After installation, many of the telepathology systems remain unused. Concerns were expressed about image quality, though mainly by pathologists who had not used the system for diagnostic work. The system remains available, but the level of use is low. CONCLUSIONS: This project has not achieved its aims. The reasons are complex, but mainly relate to human attitudes. Pathologists with excessive workloads were reluctant to use time to learn new skills which were not directed to reducing workload. IT staff did not perceive the project as part of their routine work. There were also numerous technological problems, but although image quality was cited by many, it was not a complaint of those who actively used the system. These problems have not been encountered by previous projects which involved small groups of committed enthusiasts. PMID- 12558743 TI - Breast carcinoma in women over the age of 85: distinct histological pattern and androgen, oestrogen, and progesterone receptor status. AB - AIMS: The pathogenesis of breast carcinoma in very elderly women is of interest, because oestrogen levels are likely to be extremely low during the development of the disease. In an effort to understand the pathogenesis of breast carcinoma in these women, this study was undertaken to compare the histological patterns and hormone receptor status of breast carcinomas arising in very elderly and younger women. METHODS AND RESULTS: Thirty-seven breast carcinomas from women over the age of 85 years at the time of their operation were examined histologically and compared with those from a large group of premenopausal women. The proportions of mucinous carcinoma and apocrine carcinoma were significantly greater in older women. The expression of steroid hormone receptors was studied immunohistochemically. Androgen receptor-positive carcinomas were significantly more frequent among older women, whereas progesterone receptor-positive carcinomas were significantly less frequent. There was no statistically significant difference in oestrogen receptor-alpha or -beta expression between the tumours from both groups. CONCLUSION: Breast carcinomas in women over the age of 85 years have a different morphological spectrum from carcinomas in younger age groups and may have different pathogenesis mechanisms that may be more dependent on androgen and androgen receptor interaction. Differences from the results of the other studies are discussed. PMID- 12558744 TI - Alteration of E-cadherin-mediated adhesion protein is common, but microsatellite instability is uncommon in young age gastric cancers. AB - AIMS: Gastric adenocarcinoma in young patients has been considered to differ in many ways from gastric carcinoma in older patients. This study was designed to determine the clinicopathological features and molecular mechanisms. METHODS AND RESULTS: Based on 4123 patients of gastric cancer in Seoul National University Hospital, 135 patients (3.3%) were chosen by the age of 30 years or younger. Expression of E-cadherin, beta-catenin, p53 and Epstein-Barr virus (EBV) was analysed using the tissue array method in formalin-fixed paraffin-embedded specimens and microsatellite instability (MSI) was determined. As a control, 320 cases of older patients were compared. Gastric adenocarcinoma of young patients revealed significant female predominance, type IV gross type, proximal location, diffuse type and frequent lymph node metastasis. In-situ hybridization for EBV showed higher positivity in young patients (9/78, 11.5%) than in older ones, but not statistically significant. In EBV+ cases, p53 over-expression was significantly higher in young patients than older patients (P < 0.05). Alteration of E-cadherin or beta-catenin was significantly higher in younger patients than in older patients (P < 0.05). Overall survival was significantly poorer in younger patients than older ones. The frequency of MSI was rare (1.3%, P < 0.05) in young patients compared with older patients (9.3%). CONCLUSIONS: These data indicate that gastric adenocarcinoma of young patients has a poor prognosis, possesses aggressive histopathological features, exhibits reduced expression of E cadherin and beta-catenin, and demonstrates lower MSI than tumours in older patients. PMID- 12558745 TI - Expression of cytokeratins 7 and 20 in carcinomas of the gastrointestinal tract. AB - AIMS: The differential expression of cytokeratin (CK) 7 and 20 by carcinomas may help in determining the primary site of a metastatic tumour. The aim of this study was to extend the published data on CK7 and CK20 expression in epithelial neoplasms of the gastrointestinal tract by considering the degree of differentiation and including some unusual neoplasms. METHODS AND RESULTS: Cases referred to the Armed Forces Institute of Pathology were studied prospectively for immunohistochemical expression of CK7 and CK20. Lesions from 105 patients were analysed. Adenocarcinomas of the upper gastrointestinal tract were positive for both CK7 and CK20 in 78% of cases; only poorly differentiated lesions were CK7-. Well-differentiated and moderately differentiated adenocarcinomas of the large intestine, including appendix, were CK7-/CK20+ in the great majority of cases, as were goblet cell carcinoids, but half of the poorly differentiated adenocarcinomas exhibited aberrant expression, as did most of the mixed goblet cell carcinoid/adenocarcinomas. All five high-grade neuroendocrine carcinomas were negative for both CK7 and CK20. CONCLUSIONS: Not only the site but also the grade and histological type of a gastrointestinal carcinoma should be considered when assessing cytokeratin phenotype. PMID- 12558746 TI - Small epithelial cells in human liver cirrhosis exhibit features of hepatic stem like cells: immunohistochemical, electron microscopic and immunoelectron microscopic findings. AB - AIMS: To investigate whether cells with features similar to those of the oval cells of rodents and the small epithelial cells (SEC) recently described in certain human liver diseases, i.e. hepatic progenitor cells, also occur in human liver cirrhosis. METHODS AND RESULTS: Surgical specimens from 35 cases of hepatitis B virus-positive cirrhosis (30 cases containing hepatocellular carcinoma) were investigated by immunohistochemical staining for cytokeratin 7 and albumin. Electron microscopic investigations, and immunoelectron microscopic investigations using the same antibodies and a double-labelling technique were performed in 15 and seven cases, respectively. SEC were observed in proliferated bile ductules, at the margins of regenerating nodules and in the fibrous septa in all cases of cirrhosis. The SEC were morphologically similar to the SEC described previously, and to the oval cells seen in experimental hepatocarcinogenesis. They were characterized by their small size, oval shape, scanty electron-dense or electron-lucent cytoplasm, a high nucleo-cytoplasmic ratio, tonofilaments and intercellular junctions. Immunoelectron microscopy revealed that the SEC co expressed cytokeratin 7 and albumin. Both relatively undifferentiated SEC and SEC with morphological and immunophenotypical signs of differentiation towards biliary epithelial cells and hepatocytes were found. CONCLUSIONS: SEC that exhibit morphological and immunophenotypical features of the SEC seen in certain other liver diseases are found in cirrhosis. These findings further support the hypothesis that a bipotent hepatic stem cell that may give rise to biliary epithelial cells and hepatocytes exists in the human liver. PMID- 12558747 TI - Unusual intraparenchymal growth patterns of malignant pleural mesothelioma. AB - AIMS: Malignant pleural mesothelioma is known to mimic morphologically a number of diverse reactive and neoplastic conditions. We describe three unusual intraparenchymal growth patterns of malignant mesothelioma seen in a series of 200 malignant pleural mesotheliomas. The diagnostic pitfalls associated with these findings are described and their potential medico-legal implications are highlighted. METHODS AND RESULTS: The study group comprised 200 malignant pleural mesotheliomas. In each case diagnosis was morphologically confirmed with ancillary immunohistochemistry using a broad panel of both mesothelial and epithelial markers. The patterns of intraparenchymal growth were documented and grouped as: direct subpleural; lymphangitic; and other. The 200 malignant pleural mesotheliomas comprised 118 epithelioid, 57 biphasic and 25 sarcomatoid, subtyped according to the WHO classification. Direct subpleural invasion was seen in 42 cases, lymphangitic spread in 27 cases. Other less well-defined intraparenchymal patterns included three sarcomatoid subtype malignant mesotheliomas exhibiting an intra-alveolar growth pattern mimicking epithelioid haemangioendothelioma. One epithelioid subtype malignant mesothelioma contained an intraparenchymal tumour nodule microscopically comprising lepidic spread of neoplastic cells over maintained alveolar structures mimicking bronchioloalveolar carcinoma. One epithelioid subtype malignant mesothelioma morphologically had areas in which alveoli were distended by discohesive epithelioid neoplastic cells with no interstitial invasion. The appearances mimicked desquamative interstitial pneumonia. Immunohistochemistry played an important role in the definitive diagnosis of each unusual parenchymal tumour deposit. In 126 malignant mesotheliomas no invasion of the subjacent lung parenchyma was identified. CONCLUSIONS: An awareness of the unusual parenchymal growth pattern in malignant mesothelioma is important to prevent misdiagnosis of other entities. In the medico-legal setting, the presence of epithelioid haemangioendothelioma or bronchioloalveolar carcinoma (in the absence of asbestosis) may be deemed to impact upon the patient's anticipated life expectancy and thereby would decrease the compensation settlement. PMID- 12558748 TI - 34BetaE12 expression along the whole spectrum of neuroendocrine proliferations of the lung, from neuroendocrine cell hyperplasia to small cell carcinoma. AB - AIMS: Monoclonal antibody 34betaE12 (Ck34betaE12) recognizes a set of cytokeratins (1, 5, 10, 14) expressed in normal stratified squamous epithelium. We have recently reported its expression in squamous cell carcinoma and basaloid carcinoma, in contrast to large cell neuroendocrine carcinoma, an entity with overlapping morphological features with basaloid carcinoma. We have now examined the role of Ck34betaE12 in discriminating between neuroendocrine and non neuroendocrine proliferations. METHODS AND RESULTS: We performed an immunohistochemical study of 228 cases, comprising the whole spectrum of lung neuroendocrine proliferations and tumours. All cases of neuroendocrine cell hyperplasia (n = 15), tumorlet (n = 23), typical carcinoid (n = 27) and atypical carcinoid (n = 23) were completely negative for Ck34betaE12. Although the neuroendocrine cells of small cell lung carcinoma and large cell neuroendocrine carcinoma were consistently negative, a strong and diffuse positive staining was found in the non-neuroendocrine components of combined small cell carcinoma (three of eight cases) and combined large cell neuroendocrine carcinoma (11 of 12 cases). In addition, scattered Ck34betaE12+ cells were noted in 11 of 64 (17%) large cell neuroendocrine carcinoma and in seven of 56 (12.5%) small cell carcinoma, which were not obviously histologically combined. This heterogeneity of high-grade neuroendocrine tumours was not observed in carcinoids which lack Ck34betaE12 clusters of reactive cells. There was mutual exclusion between expression of neuroendocrine markers and that of Ck34betaE12. CONCLUSION: We conclude that 34betaE12 expression excludes the neuroendocrine nature of tumour cells and uncovers the real frequency of combined forms in high-grade neuroendocrine tumours. PMID- 12558749 TI - High molecular weight cytokeratin antibody (clone 34betaE12): a sensitive marker for differentiation of high-grade invasive urothelial carcinoma from prostate cancer. AB - AIMS: There is no well-established positive immunomarker for urothelial carcinoma. We evaluated the diagnostic utility of high molecular weight cytokeratin (HMWCK) antibody clone 34betaE12 in differentiating high-grade invasive urothelial carcinoma from prostate cancer. METHODS AND RESULTS: Formalin fixed paraffin-embedded sections from 28 cases of high-grade invasive urothelial carcinoma (20 not otherwise specified (UC-NOS), eight with glandular differentiation) and 20 cases of poorly differentiated prostate carcinoma were immunostained with a monoclonal antibody to carcinoembryonic antigen (CEA), clone 85A12 and with HMWCK antibody clone 34betaE12 after microwave pretreatment or protease 24 predigestion. All cases of UC-NOS expressed HMWCK on 34betaE12 immunostaining after microwaving or enzyme predigestion. Immunoreactivity was intense and diffuse in all the cases after microwave pretreatment, whilst with enzyme predigestion immunoreactivity was sometimes patchy with <50% tumour cells positive in 20% of cases. In comparison with 34betaE12, 85A12 was insensitive with 15% of UC-NOS cases totally CEA-negative and <50% tumour cell immunoreactivity in 60% of cases. Rare positive cells were present in two (10%) cases of prostate cancer with monoclonal anti-CEA and 34betaE12 on microwaved sections, but all the cases were HMWCK-negative using 34betaE12 on sections pretreated by enzyme digestion. CONCLUSIONS: HMWCK antibody clone 34betaE12, particularly when used with microwave heat retrieval, is a very sensitive positive marker for high-grade invasive urothelial carcinoma. PMID- 12558750 TI - Splenic marginal zone atrophy and progressive CD8+ T-cell lymphocytosis in HIV infection: a study of adult post-mortem spleens from Cote d'Ivoire. AB - AIMS: Progressive changes have been reported in lymph nodes in HIV infection, but few accounts describe altered splenic histology at different stages of the disease. Investigation of splenic changes accompanying the progressive CD4+ T cell depletion that occurs in HIV infection could shed light on normal immunological interactions in this organ. Therefore, we assessed the amount and distribution of lymphoid tissue in spleens from adults with documented early or advanced HIV disease. METHODS AND RESULTS: Immunohistochemistry was used to study splenic tissue collected in an extensive autopsy survey of HIV+ adults in West Africa. Compared with post-mortem spleens from HIV- West African adults and control UK spleens, those from HIV-infected patients showed severe atrophy of white pulp B- and T-cell compartments. In early and advanced HIV disease, marginal zone atrophy was significant. Peri-arteriolar lymphoid sheaths contained increased numbers of CD8+/CD45RO+ T-cells in advanced HIV disease. In red pulp, early and advanced cases showed a lymphocytosis of CD8+/CD45RO- T-lymphocytes. CONCLUSIONS: Atrophic changes were more extreme in advanced than early HIV infection. Reduced marginal zone function possibly explains the known predisposition of HIV+ patients to infection by encapsulated bacteria. Possible immunological consequences of these CD8+/CD45RO+ (peri-arteriolar lymphoid sheaths) and CD8+/CD45RO- (red pulp) responses deserve further study. Comparison of West African and UK control spleens indicated that there were no major ethnic differences in spleen structure to prevent extrapolation of our results to European adults. PMID- 12558751 TI - High expression of markers of apoptosis in Langerhans cell histiocytosis. AB - AIMS: Langerhans cell histiocytosis is a rare disease with clonal proliferation of dendritic histiocytes, occurring most frequently in infancy and early childhood. In the localized form (single system), the disease is self-limiting, but in the cases of multisystem disease a third of the patients develop organ dysfunction. In these cases the prognosis is poor. Our objective has been to study the immunohistochemical expression of Fas and Fas-ligand (Fas-L) in order to determine whether the level of expression of these proteins could predict the outcome of the disease. We also wanted to determine the number of apoptotic cells to compare with the expression of Fas and Fas-L. METHODS AND RESULTS: We analysed the expression of Fas and Fas-L in 76 infiltrates from 49 paediatric patients with Langerhans cell histiocytosis. We also compared the results with the expression of the tumour suppressor protein p53 and the number of cells in apoptosis detected with TUNEL. Langerhans cell histiocytosis cells showed strong expression of p53 and in some cases co-expression of Fas and Fas-L. The expression of Fas-L was significantly higher in infiltrates from patients with single-system disease. The actual number of pathological Langerhans cells in apoptosis as estimated by TUNEL was low. CONCLUSIONS: The low number of TUNEL reactive cells can be explained by the rapid turnover of apoptotic cells in the tissue, not leaving the apoptotic cells long enough in the tissue to be detected. The co-expression of Fas and Fas-L in some Langerhans cells can lead to an autocrine apoptotic shortcut, mediating the death of the double-positive cells. Our findings suggest that apoptosis mediated through the Fas/Fas-L pathway may contribute to the spontaneous regression of lesions in single-system disease. A delicate balance between autocrine death and survival of Langerhans cells may have been disturbed in patients with multisystem lesions. PMID- 12558752 TI - Are acinic cell carcinoma and microglandular carcinoma of the breast related lesions? PMID- 12558753 TI - Macrophage collections in gastrointestinal biopsies and metabolic disorders: two unusual case reports. PMID- 12558754 TI - Dermatofibrosarcoma protruberans in a renal transplant recipient. PMID- 12558755 TI - Benign mixed mesenchymal tumour of the ovary (or chondrolipoleiomyoma). PMID- 12558756 TI - Aggressive adenosarcoma of the ovary. PMID- 12558760 TI - The association between migraine and juvenile stroke: a case-control study. AB - BACKGROUND: Several studies suggest an association between migraine and juvenile stroke. Because of some shortcomings, we designed another case-control study of a homogenous group of patients with juvenile cerebral ischemia. This study is part of a larger German epidemiological research project on the association of migraine with cerebrovascular disease. METHODS: We enrolled 160 consecutive patients under the age of 46 years with first-ever ischemic stroke or transient ischemic attack and 160 strictly sex- and age-matched controls. Patients suffering from arterial dissection, brain hemorrhage, cranial sinus thrombosis, lacunar stroke, or from migrainous infarction were excluded. Migraine was diagnosed according to the criteria of the International Headache Society by the same 2 independent interviewers. For analyzing the data, nonparametric statistical methods including odds ratio and 95% confidence interval were used. RESULTS: Migraine was a significant risk factor for juvenile stroke for the total sample with an odds ratio of 2.11 (confidence interval, 1.16 to 3.82). The odds ratio was even higher in the subgroup under the age of 35 (3.26) and in the female subgroup (2.68). We found migraine to be independent from other vascular risk factors, from etiology, and from the territory of stroke. CONCLUSION: We can confirm the findings of previous studies showing a significant association between migraine and juvenile stroke in women. Furthermore, our data suggest migraine to be an even more significant risk factor for patients under the age of 35 and to be independent from other vascular risk factors. PMID- 12558761 TI - Migraine aggravation caused by cephalic venous congestion. AB - BACKGROUND: Cerebral venous distension is thought by some to serve as a source of migraine pain. Previous investigators have tried to modify pain intensity by induction of additional venous congestion via compression of both internal jugular veins (Queckenstedt's maneuver). The magnitude of blood flow within the internal jugular veins depends markedly on body position, and inconsistencies in positioning may have influenced their results. OBJECTIVE: To investigate the effect of Queckenstedt's maneuver, performed both in the upright and in the supine body position, in migraineurs during an acute attack. METHODS: Twenty-five patients (18 women, 7 men; mean age +/- SD, 35.4 +/- 13.3 years) with International Headache Society-defined migraine without aura were evaluated. Queckenstedt's maneuver was performed in both body positions during an acute migraine attack, involving constant application of manual pressure to both internal jugular veins for 30 seconds. Headache intensity was rated before, during, and after Queckenstedt's maneuver on a scale extending from 1 (mild) to 10 (intolerable). RESULTS: Seventeen patients (68%) reported an increase of headache intensity in the supine position during Queckenstedt's maneuver. In the sitting position, pain increase was observed only in 6 patients (24%). The magnitude of pain increase was significantly greater in the supine position compared to the upright position (P=.02). CONCLUSIONS: Our results support a role for cerebral venous congestion in the generation of migraine pain and suggest body position may influence the clinical expression of that process. PMID- 12558762 TI - Headache and hemodialysis: a prospective study. AB - OBJECTIVE: To determine the prevalence and clinical characteristics of headache in patients undergoing hemodialysis and to compare the prevalence of headache prior to and following the beginning of a hemodialysis program. BACKGROUND: About 70% of patients receiving hemodialysis complain of headache. Despite this, headaches have not been well studied in this group of patients. METHODS: We evaluated a series of patients with chronic renal failure who attended 3 hemodialysis services in a Brazilian town from January of 1998 to December of 1999. Patients with headache were prospectively followed and classified according to the International Headache Society criteria. The headaches were also categorized according to their temporal profile relative to hemodialysis. RESULTS: Headache was reported by 87 (70.7%) of the 123 patients studied. Prior to beginning dialysis, 48% had migraine, 19% had episodic tension-type headache, and 8% had both. Headache related to arterial hypertension was the second most frequent headache diagnosis in these patients (25.4%). Fifty patients (57.5%) experienced headache during the session of hemodialysis. Thirty-four were classified as dialysis headache, 7 were classified as migraine, 7 as episodic tension-type headache, and 2 were unclassified. Twenty-four patients (27.6%) reported dramatic improvement of their headaches after the beginning of the dialysis program. CONCLUSIONS: Headache is common in patients undergoing hemodialysis. Classification of such headaches according to the International Headache society criteria may be difficult. PMID- 12558763 TI - Platelet 3H ketanserin binding in tension-type headache. AB - OBJECTIVE: The present study was undertaken to investigate the alterations in platelet 5-HT2 receptor binding in patients with tension-type headache. BACKGROUND: Serotonin (5-HT) has an important but complex role in pain modulation. The involvement of serotonin in tension-type headache has been investigated by studying serotonin in peripheral blood, but results have been inconclusive. There are, however, only a few investigations in which the status of platelet serotonin transporters has been studied by 3H imipramine and 3H paroxetine. The present study was undertaken to investigate alterations in platelet 5-HT2A receptors using 3H ketanserin as a ligand. METHODS: Platelet 3H ketanserin binding was studied in 14 patients with tension-type headache and in 15 healthy controls. The binding characteristics, equilibrium dissociation constant and maximal number of binding sites were determined by Scatchard analysis. RESULTS: There was no change in the equilibrium dissociation constant in the patients with headache as compared to the control group, but subgroup analysis revealed that patients with tension-type headache with a headache index of less than 360 had a significantly lower equilibrium dissociation constant as compared to those with a headache index of more than 360; there was a significant correlation between the equilibrium dissociation constant and the headache index. A significant decrease was observed in the maximal number of binding sites in tension-type headache. No correlation was observed between the maximal number of binding sites and age, duration of illness, or headache intensity. CONCLUSIONS: The findings of the present study show that there is a decrease in the number of binding sites of 5-HT2A receptors in some patients with tension-type headache, suggesting postsynaptic serotonergic dysfunction and the involvement of serotonin in that group. PMID- 12558764 TI - Single use of sumatriptan: a patient interview study. AB - OBJECTIVE: To investigate the possible reasons associated with the use of a single prescription of sumatriptan. BACKGROUND: A few population-based studies concerning the usage patterns of sumatriptan have revealed a relatively high incidence (approximately 40%) of sumatriptan users who utilize only a single prescription of the drug. DESIGN AND METHODS: Using automated prescription data from 11 community pharmacies, we identified single and multiple sumatriptan prescription recipients. The data were collected from May 1, 1998, to April 30, 2000. Several patient- and medication-related variables possibly associated with single recipiency of sumatriptan were analyzed. In addition, single recipients of sumatriptan were invited for an interview and asked a number of questions related to their clinical status and their experience with the medication. RESULTS: Four hundred ninety-five, first-time users of sumatriptan were identified during the patient selection period, of whom 38% were single recipients of sumatriptan. Of the latter, 102 patients were considered eligible for interview. Reasons for terminating treatment after only 1 prescription included: inefficacy and/or occurrence of side effects, 78% (n=79); uncertain diagnosis of migraine, 39.2% (n=40); and reduction in headache frequency, 33.3% (n=34). Almost half of the population had terminated treatment without having consulted their physician. More than half relied upon the use of over-the-counter (OTC) analgesics after having tried sumatriptan. Compared to multiple users of sumatriptan, single recipients were far less likely to have used another form of migraine treatment prior to (odds ratio, 0.35; [95% confidence interval, 019 to 0.67]) and after (odds ratio, 0.34 [95% confidence interval, 0.19 to 0.63]) initiating sumatriptan. Furthermore, single recipients had demonstrated an increased tendency towards benzodiazepine use prior to receiving sumatriptan (odds ratio, 1.80 [95% confidence interval, 1.00 to 3.28]). CONCLUSIONS: Single use of a sumatriptan prescription reveals some issues that may impact negatively the provision of effective migraine management. These include: rapidly developing dissatisfaction with the treatment provided and a lower tendency to seek out medical care. Our results also suggest that the drug may be used (inappropriately) as a diagnostic tool. PMID- 12558765 TI - Disability and chronic posttraumatic headache. AB - OBJECTIVE: To test the hypothesis that chronic posttraumatic headache is associated with greater severity and psychological distress than headache of nontraumatic origin. METHODS: Two hundred eighty-nine consecutive patients with chronic headache attending a university headache clinic were evaluated. Questionnaires about headache symptoms, quality of life (Medical Outcome Survey SF-36 Health Survey), and psychological distress were completed. RESULTS: Frequent headache (>4 days per week) occurred more often with traumatic (84%) than nontraumatic headache (60%). Traumatic headache resulted in greater reduced activity (3.31 +/- 1.06 days per week versus 2.62 +/- 1.11 days per week with nontraumatic headache; P<.001) and complete disability (2.89 +/- 1.17 days per week versus 2.25 +/- 1.17 days per week with nontraumatic headache; P<.001). Physical function was also rated significantly lower in patients with traumatic headache (SF-36 score: 59.8 +/- 27.7 versus 71.6 +/- 26.4 with nontraumatic headache; P<.01). These differences were not attributable to differences in headache frequency or gender between patients with traumatic and nontraumatic headache. Psychological symptoms were similar between patients with traumatic and nontraumatic chronic headache. CONCLUSIONS: Chronic traumatic headache is associated with increased headache frequency and disability compared with nontraumatic headache. Headache evaluation should include an investigation for origin of headache and consideration for more aggressive treatment in patients with traumatic headache. PMID- 12558766 TI - Progression of Tolosa-Hunt syndrome to a cavernous dural arteriovenous fistula: a case report. PMID- 12558767 TI - Nonpostural headache by spontaneous intracranial hypotension. PMID- 12558768 TI - Cluster headache after removal of a subarachnoid cyst: a case report. PMID- 12558769 TI - Familial neck-tongue syndrome. AB - Neck-tongue syndrome is an uncommon clinical entity characterized by brief attacks of intense unilateral stabbing pain in the upper neck or occipital region upon sudden rotation of the head, accompanied by ipsilateral numbness of the tongue. Eight patients, 5 teenagers and 3 adults, with neck-tongue syndrome are presented. Each of the 5 adolescents had normal examinations and normal neuroimaging. The 3 adults were parents of the affected children and had experienced transient symptoms during their adolescence suggesting an autosomal dominant inheritance pattern. PMID- 12558770 TI - Disease modification in migraine: a concept that has come of age? PMID- 12558771 TI - Ergotamine and dihydroergotamine: history, pharmacology, and efficacy. AB - Ergotamine and dihydroergotamine share structural similarities with the adrenergic, dopaminergic, and serotonergic neurotransmitters. As a result, they have wide-ranging effects on the physiologic processes that they mediate. Ergotamine and dihydroergotamine are highly potent at the 5-HT1B and 5-HT1D antimigraine receptors and, as a consequence, the plasma concentrations that are necessary to produce the appropriate therapeutic and physiologic effects are very low. The broad spectrum of activity at other monoamine receptors is responsible for their side effect profile (dysphoria, nausea, emesis, unnecessary vascular effects). Both ergotamine and dihydroergotamine have sustained vasoconstrictor actions. In acute migraine treatment, their mechanisms of action involve constricting the pain-producing intracranial extracerebral blood vessels at the 5 HT1B receptors and inhibiting the trigeminal neurotransmission at the peripheral and central 5-HT1D receptors. The scientific evidence for efficacy is stronger for dihydroergotamine than for ergotamine. Their wide use is based on long-term experience. PMID- 12558772 TI - Headache and disinhibition in an elderly woman. PMID- 12558773 TI - Your headache is a cow. PMID- 12558774 TI - Enalapril prophylaxis for migraine with aura. PMID- 12558775 TI - Guidelines on the selection and use of therapeutic products to treat haemophilia and other hereditary bleeding disorders. AB - Evidence based guidelines are presented on the selection and use of therapeutic products to treat haemophilia. These guidelines offer advice based on the best published scientific and medical information. They will be reviewed regularly by UKHCDO Advisory Committee. Included are details of therapeutic products available in the UK to treat patients with haemophilia and other bleeding disorders and the background information on which the recommendations are based are presented. PMID- 12558776 TI - Nanofiltration of plasma-derived biopharmaceutical products. AB - This review presents the current status on the use and benefits of viral removal filtration systems--known as nanofiltration--in the manufacture of plasma-derived coagulation factor concentrates and other biopharmaceutical products from human blood origin. Nanofiltration of plasma products has been implemented at a production scale in the early 1990s to improve margin of viral safety, as a complement to the viral reduction treatments, such as solvent-detergent and heat treatments, already applied for the inactivation of human immunodeficiency virus, hepatitis B and hepatitis C virus. The main reason for the introduction of nanofiltration was the need to improve product safety against non-enveloped viruses and to provide a possible safeguard against new infectious agents potentially entering the human plasma pool. Nanofiltration has gained quick acceptance as it is a relatively simple manufacturing step that consists in filtering protein solution through membranes of a very small pore size (typically 15-40 nm) under conditions that retain viruses by a mechanism largely based on size exclusion. Recent large-scale experience throughout the world has now established that nanofiltration is a robust and reliable viral reduction technique that can be applied to essentially all plasma products. Many of the licensed plasma products are currently nanofiltered. The technology has major advantages as it is flexible and it may combine efficient and largely predictable removal of more than 4 to 6 logs of a wide range of viruses, with an absence of denaturing effect on plasma proteins. Compared with other viral reduction means, nanofiltration may be the only method to date permitting efficient removal of enveloped and non-enveloped viruses under conditions where 90-95% of protein activity is recovered. New data indicate that nanofiltration may also remove prions, opening new perspectives in the development and interest of this technique. Nanofiltration is increasingly becoming a routine step in the manufacture of biopharmaceutical products. PMID- 12558777 TI - The safety and efficacy of B-domain deleted recombinant factor VIII concentrate in patients with severe haemophilia A. AB - BACKGROUND: B-domain-deleted recombinant factor VIII (BDDrFVIII) was developed when the B-domain was found to be redundant for maintaining haemostasis. This allows formulation of the final product without albumin added as a stabilizer. METHODS: Three multicentre clinical studies and one pharmacokinetic study were conducted in 218 patients to evaluate the safety and haemostatic efficacy of BDDrFVIII. RESULTS: Previously treated patients (n = 113; median duration, 1711 days; median exposure days, 385; total 98,096,287 IU infused) rated 97-99% of all infusions as good or excellent efficacy. FVIII inhibitor was noted in one patient in the previously treated patient cohort after 113 exposure days. Among 101 previously untreated patients, responses to BDDrFVIII were rated as excellent or good in 92-95% of infusions (median duration, 1413 days; median exposure days, 148; total 12,636,458 IU infused). Thirty-two previously untreated patients developed inhibitors after a median duration of 12 exposure days (range, 3-49). Sixteen of 32 (50%) patients had low levels (< or = 5 Bethesda units) and 16 had high levels of inhibitors. Inhibitors disappeared in six of 14 (43%) of the high level and six of eight (75%) of the low-level patients who underwent immune tolerance induction therapy. A total of 42 patients underwent surgery and the overall efficacy of BDDrFVIII was rated as excellent or good for 99.6% of infusions. CONCLUSIONS: The results of these clinical studies indicate that BBDrFVIII is safe and effective and has haemostatic activity similar to that of full-length FVIII concentrates. PMID- 12558778 TI - Complications of central venous access devices in paediatric haemophilia patients. AB - We conducted a retrospective survey of our experience with central venous access devices (CVADs) implanted in children with haemophilia seen at the Vanderbilt Hemostasis-Thrombosis Clinic from 1986 to 2000. Following discussion with parents on the merits and risks associated with the use of CVADs for immune tolerance induction or factor prophylaxis, catheters were inserted under sterile technique in the operating room. One nurse provided demonstration and teaching about catheter care and access. Thirty central venous catheters were inserted in 22 children. Our survey revealed that the two most common complications associated with central venous catheters were bacteraemia and thrombosis. We found a sepsis rate of 0.30/1000 catheter-days or one episode of bacteraemia for every 3346 days of catheter use. The thrombosis rate of our cohort was 0.13/1000 catheter-days or one episode of thrombosis for every 7529 days of catheter use. Uncomplicated venous access is essential in children with severe haemophilia who require prophylaxis or immune tolerance induction. While infection was the most common complication observed in our series, we experienced a lower overall infection rate than several reported series. Catheter thrombosis and subsequent obstruction may occur as a result of intraluminal fibrin deposits. We conclude that the use of implantable central venous catheters is an effective method for accessing children with haemophilia. We accept that the benefits of CVADs in the treatment of paediatric haemophilia patients outweigh the previously documented risks. Future prospective studies should be designed to define all associated risks and to determine effective strategies to reduce them. PMID- 12558779 TI - An evaluation of the stability of factor VIII inhibitors in plasma and plasma dried on filter paper discs stored at room temperature. AB - The transportation of plasma specimens to specialized haemophilia centre laboratories for anti-factor VIII inhibitor titre determination is often necessary. The routine method of transporting frozen specimens on dry ice is limited by its cost and need for special handling. The present study was undertaken to evaluate the effect of storing specimens at room temperature on the FVIII inhibitor titre determinations using the Bethesda assay. Specimens stored both in liquid phase as well as adsorbed onto filter paper discs were studied. The results of the present study demonstrate that plasma specimens stored for up to 2 weeks at room temperature, either in liquid phase or adsorbed onto filter paper, yield equivalent measures of FVIII inhibitor titres using the Bethesda assay to plasma specimens stored frozen at -70 degrees C. Plasma specimens dried on filter paper discs and stored at room temperature offers a reliable, more practical and less expensive alternative to frozen plasma as a means of transport to specialized referral laboratories for analysis of anti-FVIII titres. PMID- 12558780 TI - Intra-articular rifamycin in haemophilic arthropathy. AB - Synoviorthesis is the intra-articular injection of chemical or radioactive substances able to produce fibrosis of hypertrophied synovium, which has proved effective in the treatment of chronic haemophilic synovitis. Between September 1999 and October 2001 we treated 28 outpatients (25 with haemophilia A, three with haemophilia B). Our treatment was focused on pain and functional limitation of joints. A schedule was adopted to treat each joint using intra-articular rifamycin once a week, repeated five times. Patients were covered with factor replacement on demand. Oral analgesia was offered as required because of acute but transient painful inflammatory reaction. Their median age was 34 years (range 15-60 years). The indication for synoviorthesis was chronic synovitis characterized by recurrent haemarthroses, persistent pain and limited range of motion (ROM). Thirty-five joints were treated with a total of 169 injections, including six joints (20%) in patients with inhibitors. In five patients two joints were treated in the same session. Thirty procedures were completed: 24 (80%) were considered effective (as excellent or good), while six were considered insufficient (20%). Pain was reduced in 96% of cases and in 70% the ROM was improved. In our experience intra-articular infiltration with rifamycin appears to be effective in reducing joint pain and in improving the ROM. The procedure presents a low risk of bleeding, can be used for patients with inhibitors and multiple joints can be treated without any additional cost. PMID- 12558781 TI - Treatment of chronic haemophilic synovitis in humans with D-penicillamine. AB - Chronic proliferative synovitis secondary to haemathroses is a major complication in patients with severe haemophilia. Current management strategies include prophylactic infusions of the missing coagulation factor, corticosteroids, synoviorthesis and/or synovectomy with variable degrees of benefit. In addition, patients with coagulation factor inhibitors are not amenable to the invasive therapeutic modalities. The gross and microscopic findings of the synovitis in haemophilic arthritis are remarkably similar to those seen in patients with rheumatoid arthritis, although the pathophysiology of these two conditions are quite different. Haemophilic arthropathy, in the later stages, resembles degenerative rather than inflammatory joint disease. Oral D-penicillamine, a drug effective in the proliferative synovitis of rheumatoid arthritis, was evaluated in 16 patients. Ten patients had an unequivocal response, while three had a reduction in palpable synovium and three had no response. Thus 81% of the patients had a beneficial response. Minor reversible drug side-effects occurred in two patients (proteinuria in one and a rash in the second). The results of this study suggest that D-penicillamine is an effective and safe drug for the treatment of haemophilic chronic synovitis. PMID- 12558782 TI - Long-term follow-up after osteotomy for haemophilic arthropathy of the knee. AB - In this study the long-term value of corrective osteotomy around the knee was evaluated by means of clinical and radiographic parameters. Between 1974 and 1984 we performed 52 corrective osteotomies in the vicinity of the knee on patients affected by haemophilic arthropathy. Forty-two patients (45 osteotomies) were adequately followed-up at an average 11.6 years postoperatively. Using the clinical score of the Advisory Committee of the World Federation of Haemophilia, 38 patients showed a postoperative improvement, five remained clinically unchanged and two showed deterioration. Range of motion of the knee joint did not significantly improve postoperatively. The radiographic Pettersson score showed only a marginal decrease by an average 0.003 points at the time of follow-up. Only one patient needed subsequent joint replacement of both knees, on the left side 13 years after osteotomy and on the right side 8 years after osteotomy. Even in cases of marked radiographic joint destruction, corrective osteotomy shows acceptable long-term clinical results, underlining the feasibility of this management option in the treatment of haemophilic arthropathy of the knee. Although moderate cartilage degenerations in the femoropatellar complex and in the contralateral compartment can be tolerated, this therapy should primarily be contemplated for those patients where damage is unicompartmental and a corresponding axial deviation is found. Particularly the younger patient can benefit from this treatment option in that joint replacement may possibly wholly be avoided or at least postponed to a later stage of life. PMID- 12558783 TI - Value of magnetic resonance imaging for the diagnosis and follow-up of haemophilic arthropathy. AB - An orthopaedic assessment of the joint status of seven severe haemophiliacs (mean age 12.5 y - range 8.9-15.5) on prophylactic treatment (PT) by magnetic resonance imaging (MRI) and physical examination was carried out. Median duration of PT of these patients was 84 months (range 32-107). A locally designed MRI joint score considering seven parameters (1 - joint effusion, 2 - synovial membrane thickening, 3 - haemosiderin deposits, 4 - joint cartilage injury, 5 - subchondral bone erosion, 6 - subchondral bone cysts, 7 - osteonecrosis) was used (maximum score = 13 points). MRI scans were performed in 17 joints (nine ankles, seven knees and one elbow): mean MRI scores for the affected joints was 5.1. A poor consistency between physical examination and MRI findings or the real extent of joint damage was found (Cohen kappa index 0.320). MRI is a precise non invasive tool for the assessment of early joint cartilage and synovium pathological changes still undetectable by physical examination or conventional X rays in the haemophilic setting. PMID- 12558784 TI - Physical training increases isometric muscular strength and proprioceptive performance in haemophilic subjects. AB - Sufficient muscular strength and proprioception lessen the risk of joint damage, however, both are impaired in haemophilic subjects. The aim of the study was to investigate proprioceptive performance and isometric muscular strength before and after a specialized training in haemophilic subjects (H) compared with two groups of control subjects (C). Nine subjects with severe haemophilia A, and eight 'active' C (AC) without haemophilia took part in a physical training programme over a 6-month period. Eleven 'passive' C (PC) were requested to avoid any additional training during this period. Proprioceptive performance and isometric strength were determined before and after the training programme. The maximal isometric muscular strength in the legs, bilaterally measured by knee extensor (and leg press) was increased (P < 0.05) by 34% (29%) after training in the H and by 20% (28%) in the AC groups while remaining unchanged in the PC group. The performance in one-leg-stand tests after training was increased (P < 0.05) in the H and AC groups. An improvement of angle reproduction of 20 degrees and 40 degrees (P < 0.05) in the H compared with the PC groups was seen in the tests. Quantitative sensory testing by the tuning fork showed an increase (P < 0.05) in performance of both H and AC groups. The results of the present study confirm that specific sports therapy focused on proprioceptive function and accompanied by gentle strength training with low resistance and 20-25 repetitions is able to increase proprioceptive performance and muscular strength with a minimal stress to the joints. It is strongly recommended that specialized sports therapy be included as an integral component of the complete treatment regimen of haemophilic subjects. PMID- 12558785 TI - Bone marrow mesenchymal cells for haemophilia A gene therapy using retroviral vectors with modified long-terminal repeats. AB - Bone marrow (BM) cells are attractive target cells for ex vivo gene therapy of genetic diseases, including haemophilia A. However, BM-derived haematopoietic stem/progenitor cells (HSCs) transduced with factor VIII (FVIII) retroviral vectors, failed to express FVIII in vivo. To overcome the limitations of HSCs for haemophilia gene therapy, BM-derived mesenchymal cells were explored as alternative target cells. The BM mesenchymal cell population contains self renewing mesenchymal stem/progenitor cells that give rise to different mesenchymal lineages and have been used safely in phase I gene-marking trials. Human BM mesenchymal cells were transduced in vitro with an improved retroviral vector encoding a human B-domain deleted FVIII (hFVIIIdeltaB) cDNA (MND-MFG hFVIIIdeltaB). This vector contains multiple modifications in the cis-acting elements within the MoMLV long-terminal repeats (LTR) that prevent the binding of repressive transcription factors. These modifications were previously shown to increase and prolong gene expression in embryonic stem (ES) cells and HSCs. Transduction of BM mesenchymal cells with the MND-MFG-hFVIIIdeltaB retroviral vector resulted in high levels of functional human FVIII in vitro, ranging between 300 +/- 50 SD and 700 +/- 100 SD mU per 106 cells per 24 h. Following xenografting of the transduced human BM cells into immunodeficient NOD-SCID mice, therapeutic hFVIII levels of 12 +/- 10 ng mL-1 were detected in the plasma. Polymerase chain reaction analysis demonstrated long-term engraftment (>3 months) of the human BM mesenchymal cells. The long-term persistence of BM mesenchymal cells in the absence of myelo-ablative conditioning and the therapeutic FVIII levels in vivo underscore the potential usefulness of BM-derived mesenchymal cells for haemophilia gene therapy, as opposed to BM-derived HSCs. Despite the modifications of the MoMLV LTR, FVIII expression declined, which coincided with a decrease in FVIII mRNA transcription levels, indicating that the salutary effect of the LTR modification on transgene expression is not universally applicable to all cell types. PMID- 12558786 TI - Optimization of storage conditions for diluted working solutions of porcine factor VIII and performance of the Bethesda assay for the determination of antiporcine FVIII inhibitor titres. AB - The use of porcine factor VIII (FVIII) (Hyate:C, Ipsen) has proven to be very successful in treating patients with FVIII inhibitors. The best way to predict the usefulness of porcine FVIII therapy, and/or to estimate the appropriate treatment dose in a given patient, is to measure the patient inhibitor titre against porcine FVIII with the Bethesda assay, using porcine FVIII as the source of FVIII in the assay. The goals of the present study were to (1) find the optimal storage temperature, diluent and concentration for a working solution of porcine FVIII to be used as the source of FVIII for the porcine Bethesda assay, (2) assess the reliability of the labelled FVIII units in the preparation of such working solutions of porcine FVIII and (3) compare the inhibitor titres determined by the Bethesda assay using both porcine and human standard reference curves for measuring residual FVIII. The results of the present study demonstrate that a ready-to-use working solution of 1 U mL(-1) of Hyate:C diluted in human FVIII deficient plasma, either containing or deficient in von Willebrand factor antigen, is stable for up to 12 months, at -20 degrees C. The preparation of the 1 U mL(-1) working solution could be reliably calculated based on the units indicated on the vial label. Finally, using the human standard curve yields similar results to using the porcine standard curve for measuring any titre of allo- or auto-antibody against FVIII in the Bethesda assay, using Hyate:C as the source of FVIII. These findings are of practical value when performing a porcine FVIII-based Bethesda assay. PMID- 12558787 TI - Hepatitis G virus in clotting factor concentrates. AB - Blood-borne hepatitis is a well-known complication in patients with bleeding disorders. A recently discovered parentally transmitted virus, hepatitis G [GB virus C (GBV-C)] has an increased prevalence in patients with haemophilia. Clotting factor concentrates derived from pools of human plasma currently undergo viral inactivation techniques known to be effective against hepatitis B, C and HIV; however, the effectiveness of current purification and viral inactivation techniques against newly discovered viruses such as GBV-C is unknown. A total of 37 vials of clotting factor concentrates manufactured in the USA from 1981 to 1995 were tested for the presence of GBV-C virus. All samples that did not undergo a specific viral inactivation step were positive for GBV-C. Viral inactivation techniques that did not uniformly remove GBV-C included vapour heat treatment and dry heat treatments for less than 144 h. All samples treated by pasteurization, solvent detergent or dry heat for 144 h, were negative for the presence of GBV-C. PMID- 12558788 TI - Carrier testing and prenatal diagnosis of haemophilia B by SSCP in an Iranian family. AB - Haemophilia B is an X-linked recessive bleeding disorder caused by multiple molecular defects in factor IX gene. During our genotyping programme of haemophilia B patients, a family from the west of Iran was referred to us. We first detected the mutation in the index case of the family by single-strand conformation polymorphism (SSCP) technique. This technique was then applied in carrier detection and prenatal diagnosis for females of this family. Sequencing later revealed the mutation as being G to A at 20519. PMID- 12558789 TI - A case of an inhibitor autoantibody to coagulation factor VII. PMID- 12558790 TI - Surgery in severe factor XIII deficiency: report of a case of epilepsy neurosurgery and review. AB - Factor XIII (FXIII) deficiency is a rare autosomal recessive congenital disorder of haemostasis, associated with a high risk of intracranial haemorrhage. Intracranial haemorrhage can result in neurological sequelae including seizure disorders. In some cases, medically intractable epilepsy led to epilepsy surgery. Little has been reported on the management of FXIII deficiency during surgery, and there is only a few data on the management, safety and efficacy of epilepsy surgery in the patients with haemostatic disorder. We report here an epilepsy neurosurgery in a case of severe FXIII deficiency. PMID- 12558791 TI - Female haemophiliac homozygous for the factor VIII intron 22 inversion mutation, with transcriptional inactivation of one of the factor VIII alleles. AB - Phenotypic expression of X-linked recessive disorders, including haemophilia A, is rare in females. This report describes a female with sporadic severe haemophilia A. The female patient and her family members were evaluated by coagulation assays. Visible detectable disturbance of X chromosome structure or number, as well as 2N von Willebrand disease, were excluded as possible explanations of the haemophilia A phenotype. Molecular studies, factor VIII (FVIII) intron 22 inversion mutation analysis showed that the severe haemophilia A phenotype is the result of a maternally inherited, distal, FVIII gene inversion and a paternally inherited de novo, also distal, FVIII gene inversion. Furthermore, comparative single-stranded conformation polymorphism analysis revealed the absence of detectable maternally inherited abnormal FVIII gene transcript in the patient's peripheral blood lymphocytes. X chromosome methylation analysis indicates that this could be explained by preferential inactivation of the maternally inherited X chromosome carrying the distal FVIII gene inversion. PMID- 12558792 TI - Efficacy of NovoSeven during surgery on a haemophiliac with previous history of inhibitors. AB - Management of bleeding in haemophiliacs with a history of inhibitor remains problematic. With infusion of factor VIII (FVIII), development of an anamnestic response and possible appearance of high-titre inhibitor remains a valid concern. We report a case of a haemophiliac with a history of moderately high-titre FVIII inhibitor that had become undetectable. He had not received FVIII since 1997, when he became inhibitor negative. He had been managed during his bleeding episodes with prothrombin complex factor concentrates, which became less effective in controlling his bleeding. The patient had a history of recurrent, spontaneous shoulder joint dislocations with bleeding, pain and significant disability. Shoulder joint replacement surgery was suggested. Replacement therapy was discussed with the patient, who refused treatment with human FVIII because of his concern for possible anamnestic response and inhibitor rebound. Porcine FVIII was not acceptable due to his poor response when used once in the past, and his history of moderate allergic reaction. Therefore, recombinant factor VIIa (NovoSeven, Novo Nordisk, Princeton, NJ) was considered to be an acceptable option for the contemplated shoulder surgery. The patient underwent 2.5 h of surgery with NovoSeven infusion. The surgeons were impressed with the lack of bleeding in this traumatic surgery. Despite the continuously prolonged activated partial thromboplastin time and low FVIII levels, the patient maintained a remarkably dry surgical field. Effective haemostasis was achieved during and after this procedure. This case illustrates the usage of NovoSeven as an effective treatment modality in a haemophilia A patient with past history of inhibitor undergoing joint surgery. PMID- 12558793 TI - Type 2B vWD: the varied clinical manifestations in two kindreds. AB - Type 2B von Willebrand's disease (vWD) is associated with spontaneous binding of large von Willebrand factor (vWF) multimers to platelets in vivo, followed by clearance of both the large multimers and platelets resulting in thrombocytopenia, which may be intermittent, mild to severe, and may be exacerbated by stress such as infection or pregnancy. We report our experience in two kindreds (49 caucasian individuals) with type 2B vWD and discuss their varied clinical manifestations. The largest kindred (45 patients) was traced back five generations to a presumed index case. The genetic defect in this kindred was identified as a missense mutation, with a C to T transition at a CpG dinucleotide (nucleotide 3916) resulting in an amino acid substitution (Arg 543 to Trp) within the glycoprotein Ib binding domain of vWF. Ristocetin cofactor activity varied from < 10 to 28%, and factor VIII activity from 7 to 69%. Analysis of von Willebrand multimers consistently revealed loss of large molecular weight forms. Platelet counts in those with thrombocytopenia varied from 10 x 10(9) L(-1) to 120 x 10(9) L(-1). The severity of thrombocytopenia has also varied within the same individual during the period of follow-up. The clinical manifestations were varied and ranged from mild to moderate spontaneous bleeding episodes, including epistaxis, menorrhagia and gastro-intestinal haemorrhage. Severe bleeding episodes were observed in those undergoing surgery (both elective and non elective), and in a few patients despite aggressive replacement with exogenous source of intact vWF, antifibrinolytics when indicated, and a near normal platelet count. Thrombotic disease may be a rare and unusual sequela of this disorder as was noted in one of our patients. Obtaining a platelet count at birth in infants of mothers with type 2B vWD who exhibit thrombocytopenia, may help in the earlier detection of infants at risk for thrombocytopenia. PMID- 12558794 TI - Diversity of the cadherin-related neuronal receptor/protocadherin family and possible DNA rearrangement in the brain. AB - Both the brain and the immune systems are complex. The complexity is generated by enormously diversified single cells. In the immune system, extensive cell death, gene regulation of immunoglobulin (Ig) and T-cell receptor (TCR) gene expression, and somatic rearrangement and mutations are known to generate an enormous diversity of lymphocytes. In this process, double-strand DNA breaks (DSBs) and DSB repair play significant roles. These processes at a DNA level are also physiologically significant in the nervous system during neurogenesis, and chromosomal variations have been detected in the nucleus of differentiated neurones. In another parallel with the immune system, cadherin-related neuronal receptors (CNRs) are diversified synaptic proteins. The CNR genes belong to protocadherin (Pcdh) gene clusters. Genomic organizations of CNR/Pcdh genes are similar to that of the Ig and TCR genes. Somatic mutations in and combinatorial gene regulation of CNR/Pcdh transcripts during neurogenesis have been reported. This review focuses on the diversity of the CNR/Pcdh genes and possible DNA diversification in the nervous system. PMID- 12558795 TI - Thrombopoietin-induced CXC chemokines, NAP-2 and PF4, suppress polyploidization and proplatelet formation during megakaryocyte maturation. AB - BACKGROUND: We previously reported that the expressions of two CXC chemokines, neutrophil activating peptide-2 (NAP-2) and platelet factor-4 (PF-4), were induced by megakaryocyte-specific cytokine thrombopoietin (TPO) in mouse bone marrow megakaryocytes. The roles of these chemokines on megakaryocyte maturation/differentiation processes, including polyploidization and proplatelet formation (PPF) remain unresolved. RESULTS: NAP-2 and PF-4 suppressed the PPF of mature megakaryocytes freshly prepared from mouse bone marrow as well as that of the megakaryocyte progenitors, c-Kit+CD41+ cells, isolated from mouse bone marrow and cultured with TPO. NAP-2 and PF-4 inhibited polyploidization of c-Kit+CD41+ cells in the presence of TPO, and also inhibited the proliferation of c-Kit+CD41+ cells. CONCLUSIONS: NAP-2 and PF-4 produced by TPO stimulation in megakaryocytes suppress megakaryocyte maturation and proliferation as a feedback control. PMID- 12558796 TI - Taxilin; a novel syntaxin-binding protein that is involved in Ca2+-dependent exocytosis in neuroendocrine cells. AB - BACKGROUND: The syntaxin family is a central coordinator and participates in multiple protein-protein interactions in the soluble N-ethyl maleimide-sensitive factor attachment protein receptor machinery, which is involved in intracellular vesicle traffic. However, the molecular mechanism by which the syntaxin family regulates intracellular vesicle transport is not well known. RESULTS: We have identified and purified a novel binding partner of syntaxin-3 from rat lung, and isolated and sequenced the cDNA of its human homologue from a human brain cDNA library. The cDNA had an open reading frame encoding a protein of 546 amino acids with a calculated Mr of 61,890. We tentatively referred to this protein as taxilin. A structural analysis of taxilin revealed the existence of an extraordinarily long coiled-coil domain in its C-terminal half. Syntaxin-1a and 4, as well as syntaxin-3 interacted with taxilin, but syntaxin-7 or -8 did not. Northern blot analysis showed that taxilin was ubiquitously expressed. Over expression of full-length taxilin inhibited Ca2+-dependent exocytosis in PC12 cells. CONCLUSIONS: These results indicate that taxilin is a novel binding partner of several syntaxin family members and suggest that taxilin is involved in Ca2+-dependent exocytosis in neuroendocrine cells. PMID- 12558797 TI - Cytoskeletal reorganization induced by insulin: involvement of Grb2/Ash, Ras and phosphatidylinositol 3-kinase signalling. AB - BACKGROUND: Cytoskeletal reorganization is important for a wide variety of insulin-mediated biological actions, including cell growth, migration and metabolism, but the intracellular signalling pathways leading to insulin-induced cytoskeletal reorganization have largely been unknown. We therefore investigated the involvement of Grb2/Ash-Ras and phosphatidylinositol (PI) 3-kinase in the insulin-induced morphological changes in fibroblasts over-expressing human insulin receptors (HIRcB cells). RESULTS: Insulin, as well as 12-O tetradecanoylphorbol-13-acetate (TPA) and 8-bromo-cAMP, induced a unique morphological change associated with actin cytoskeletal reorganization characterized by the disruption of actin stress fibres and thicker actin bundle formation. Microinjection of an anti-Grb2/Ash antibody, but not control IgG, inhibited the insulin-induced actin reorganization, whereas the TPA- and 8-bromo cAMP-induced morphological changes were not inhibited by microinjection of the anti-Grb2/Ash antibody. In addition, microinjection of dominant negative ras p21 protein, but not the heat-treated protein, inhibited insulin-induced cytoskeletal reorganization. Microinjection of activated p21ras protein resulted in very similar cytoskeletal reorganization with actin bundle formation in the cytoplasm. The PI3-kinase inhibitor wortmannin inhibited insulin-induced cytoskeletal reorganization, but not the TPA- nor 8-bromo-cAMP-induced reorganization. Interestingly, wortmannin also inhibited the activated p21ras-induced morphological change. CONCLUSIONS: We concluded that Grb2/Ash-Ras activation and probably Ras-associated PI3-kinase activation are involved in the insulin-induced morphological change. PMID- 12558798 TI - Artificial recruitment of certain Mediator components affects requirement of basal transcription factor IIE. AB - BACKGROUND: Basal transcription factors are essential for RNA polymerase II (RNAPII)-catalysed transcription of many but not all the mRNA-encoding genes in vivo as well as in vitro. For example, copper-inducible transcription of the copper metallothionein gene CUP1 occurs independently of basal factor TFIIE in budding yeast. To gain insight into the mechanism by which the requirement for TFIIE is bypassed, we artificially recruited certain constituents of Mediator, a large protein complex transmitting signals from various activators to the RNAPII machinery, to the CUP1 promoter by protein fusions with Ace1, the copper inducible activator. RESULTS: Fusions with Med2 or Pgd1 activated CUP1 independently of TFIIE. Surprisingly, fusions with neither Srb5 nor Med9 circumvented TFIIE requirement for the CUP1 activation. Components of TFIID were similarly recruited to the CUP1 promoter without activation. By using a chromatin immunoprecipitation technique, we found that TFIIE is necessary for stable binding of TFIIH and RNAPII to the ADH1 promoter, whose activation requires TFIIE. However, binding of TFIIH and RNAPII to CUP1 upon its activation did not require TFIIE. CONCLUSIONS: Our results strongly suggest that the TFIIE requirement of a gene is determined by a target(s) in Mediator through which the signal of the cognate activator is transmitted. PMID- 12558799 TI - Antagonistic and agonistic effects of an extracellular fragment of nectin on formation of E-cadherin-based cell-cell adhesion. AB - BACKGROUND: Nectin is a Ca2+-independent immunoglobulin-like cell-cell adhesion molecule at the E-cadherin-based cell-cell adherens junctions (AJs), and comprises a family consisting of four members, nectin-1, -2, -3, and -4. Nectin and E-cadherin are associated with afadin and alpha-catenin, actin filament (F actin)-binding proteins connecting respective adhesion molecules to the actin cytoskeleton, but the role of nectin in the formation of the E-cadherin-based cell-cell AJs has not yet been fully understood. To obtain evidence for this role of nectin, we attempted to develop an antagonist and/or agonist of nectin. RESULTS: We made a recombinant extracellular fragment of nectin-3 (Nef-3). Nef-3 trans-interacted with cellular nectin-1 and thereby diminished the formation of the nectin-1-based cell-cell adhesion. This resulted in a reduction of the formation of the E-cadherin-based cell-cell adhesion in L fibroblasts stably expressing both exogenous nectin-1alpha and E-cadherin (nectin-1-EL cells) and MDCK cells stably expressing exogenous nectin-1alpha (nectin-1-MDCK cells). This antagonistic effect of Nef-3 was also observed in L cells stably expressing exogenous E-cadherin alone (EL cells) and wild-type MDCK cells. Conversely, Nef-3 coated on microbeads first recruited the nectin-afadin complex and then the E cadherin-catenin complex to the bead-cell contact sites in nectin-1-EL and nectin 1-MDCK cells. CONCLUSION: These results suggest that nectin is necessary and sufficient for the recruitment of E-cadherin to the nectin-based cell-cell adhesion sites and involved in the formation of E-cadherin-based cell-cell AJs. PMID- 12558800 TI - A possible linkage between AMP-activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) signalling pathway. AB - BACKGROUND: The mammalian target of rapamycin (mTOR) regulates multiple cellular functions including translation in response to nutrients, especially amino acids. AMP-activated protein kinase (AMPK) modulates metabolism in response to energy demand by responding to changes in AMP. RESULTS: The treatment of SV40 immortalized human corneal epithelial cells (HCE-T cells) with 5-aminoimidazole-4 carboxamide ribonucleoside (AICAR), widely used as an AMPK activator, inhibits p70 S6k activities. Altered glucose availability, which regulates AMPK activity, also modulates the activity of p70 S6k. AICAR treatment also inhibits phosphorylation of Thr-412 in the p70 S6 kinase (p70 S6k), which is indispensable for the activity. Furthermore, over-expression of mutant AMPK subunits by stable expression in rabbit pulmonary fibroblast cell lines (PS120 cells) also modulates p70 S6k activity. The insensitivity of the rapamycin-resistant p70 S6k variant to AICAR treatment suggests that the inhibition of p70 S6k is mediated through a common effector, supporting a model whereby mTOR and its downstream effector are controlled by AMPK. CONCLUSION: These results indicate that the AMPK and mTOR signalling pathways are possibly linked. In addition to the mTOR signal acting as a priming switch that modulates p70 S6k activation, AMPK appears to provide an overriding switch linking p70 S6k regulation to cellular energy metabolism. PMID- 12558801 TI - Defining and classifying periodontitis: need for a paradigm shift? AB - The past two decades have witnessed a large number of proposals for the classification of periodontitis. These proposals are all founded in an essentialistic disease concept, according to which periodontitis is a link between the causes and the signs and symptoms of periodontitis. Essentialistic definitions are necessarily rather imprecise and thereby subject to multiple interpretations. Consequently, it remains unknown to what extent current knowledge regarding 'different' forms of periodontitis is based on the 'same' type of patients. However, periodontitis is a syndrome, the clinical manifestations of which may come in all sizes. Thereby, periodontitis has no diagnostic truth, just as there is no natural basis for a sharp distinction between health and disease or between 'different' forms of periodontitis. Recognition of these facts and adoption of a nominalistic approach to the definition of periodontitis is needed to provide a rational framework for the development of a classification system that meets the needs of both clinicians and scientists. PMID- 12558802 TI - In vitro study of caries detection through sound dentin using a laser fluorescence device, DIAGNOdent. AB - The objective of this study was to investigate the feasibility of proximal surface caries detection using a laser fluorescence device, DIAGNOdent, through the sound dentin surrounding the cavity wall after removal of occlusal caries. Extracted sound human molars were ground to prepare horizontal and vertical dentin plates 1.4 mm thick. Extracted carious human molars were reduced in thickness horizontally from the occlusal surfaces until exposure of the dentin caries. The dentin plate was placed on the carious tooth. Before and after gradual thinning of the dentin plate from approximately 1.4 mm to 0.2 mm thick, the dentin caries was measured 10 times through the plate by the laser fluorescence device with a cone-shaped tip or a broad tip. When the dentin plate was reduced to less than 0.3 mm thick (using a combination of a horizontal plate and cone-shaped tip) or 0.2 mm thick (using the other combinations), the values measured with the laser fluorescence device were significantly larger than the values before reducing the thickness of the dentin. Based on these results, the device offers potential use as one of the screening tests for proximal surface caries detection through sound dentin when the sound dentin between tip and caries is thin. PMID- 12558803 TI - The Fear of Dental Pain questionnaire: construction and validity. AB - Anxiety, fear and pain play an important role in the context of dental treatment and patients' well-being. The Fear of Pain Questionnaire (FPQ)-III is a recently developed self-report questionnaire measuring pain-related fear of a variety of painful stimuli. The present study was undertaken in order to develop a dental equivalent of the FPQ-III, called the Fear of Dental Pain questionnaire (FDP), to determine standard psychometric characteristics and to assess the instruments' validity. Four experienced dentists generated the initial pool of items and two methodologists constructed the initial questionnaire. Two studies were performed. In the study one, a sample of psychology freshmen (n = 309) was taken in order to analyse response patterns. In study two, a sample (n = 176) of patients, dental students and the general population was examined. Results from both studies were used to determine reliability and validity. High internal consistency (0.93) with satisfactory test-retest reliability (0.75) was obtained. Factor analysis revealed a strong one-dimensional factor underlying almost all items. Finally, the proposed FDP version was related to a measure of dental fear and a general measure of fear of pain. All a priori hypotheses were confirmed, thereby providing evidence for the validity of the FDP. The FPD may prove to be a clinically useful tool in the dental setting, and a potentially important covariate in dental pain perception research. PMID- 12558804 TI - Living with a child with a severe orofacial handicap: experiences from the perspectives of parents. AB - Orofacial functions include competences/abilities such as eating, breathing, speech/language, mimicry, as well as oral health, and disturbances are common in children with rare disorders. To describe parental experiences of orofacial function and needs in children with rare disorders, in-depth interviews focusing on orofacial function were carried out with 14 parents. Interviews were transcribed verbatim and analysed in open, axial (theoretical) and selective coding processes according to Grounded Theory. Two core categories emerged from data in the analysis: 'the vulnerable family' and 'support perceived from others'. The data indicated the importance of a balance between these two core categories: the strain caused by living in a family with a child with a severe disability/handicap, and the availability of perceived support from caregivers and significant others. This balance was necessary for the parents in developing self-reliance and in reconciling themselves to their life situations. Parents described orofacial dysfunction in terms of feeding and communication problems, needs for orthodontic treatment to reduce risk of trauma or improve chewing, and drooling. Oral health issues such as dental caries and gingivitis were not mentioned. Five aspects of good professional attitude were recognized: respect, involvement, continuity, knowledge, and availability. PMID- 12558805 TI - Immunohistochemical localization of insulin-like growth factor-II and its binding protein-6 in human epithelial cells of Malassez. AB - So-called epithelial rests of Malassez are derived from the Hertwig's root sheath and are located in the periodontal ligament, with still unknown functions. Different pathological conditions may lead to proliferation of these otherwise non-proliferative cell clusters. The insulin-like growth factor (IGF) system is an important growth factor system controlling proliferation and differentiation. In our study on Malassez cells from extracted human deciduous teeth, we investigated their structure by means of light and electron microscopy. Although they appeared as cellular clusters with a uniform epithelial phenotype, immunohistochemical analyses of components of the IGF system revealed an unique pattern: weak immunoreactivity could be seen for IGF-II while among all IGF binding proteins (IGFBPs) only IGFBP-6 and weakly IGFBP-4 were detectable in epithelial cells of Malassez. Since IGFBP-6 has a very high affinity for IGF-II and can inhibit its functions, we discuss that, in the normal periodontal ligament, autocrine IGFBP-6 may function as an antiproliferative molecule suppressing mitogenic effects of IGFs on Malassez cells. PMID- 12558806 TI - Analysis of proliferation, apoptosis and keratin expression in cultured normal and immortalized human buccal keratinocytes. AB - The current study was undertaken to analyse growth and differentiation-related functions of normal keratinocytes (NOK) and an SV40T-immortalized keratinocyte line (SVpgC2a) from buccal mucosa, viewing the latter cell line as a model of a dysplastic epithelium. Morphological and immunohistochemical assessments of organotypic epithelia generated from 10 or 17 d of culture showed three- to five fold higher apoptotic and proliferative activity in SVpgC2a relative to NOK. Conditions with or without serum (up to 10%) did not significantly influence these parameters in NOK whereas serum supported proliferation of SVpgC2a. Both cell types showed basal expression of collagen IV and laminin 1, indicating basal lamina, as well as vimentin, indicating an activated, proliferative state. Reduced expression of keratin, including the non-keratinizing marker K13, was seen in SVpgC2a. Assessment of proliferative monolayer cultures by microarray showed that NOK transcribed tissue-specific keratins, but also the epidermal keratin K2a, several simple epithelial keratins and low levels of hair keratins. SVpgC2a transcribed keratins seen in epithelial dysplasia, and K2a and hair keratins, albeit at low level. Overall, the results implied aberrant apoptosis, proliferation and keratin expression in the immortalized state of SVpgC2a. Comparison of NOK and SVpgC2a under identical culture conditions may serve to model the progression from a normal to a pre-neoplastic state of buccal epithelium. PMID- 12558807 TI - Keratinocyte growth factor and scatter factor expression by regionally defined oral fibroblasts. AB - Keratinocyte growth factor (KGF) and hepatocyte growth factor/scatter factor (SF) are two signalling molecules thought to play important roles in regulating epithelial-mesenchymal interactions. Expression of both factors by fibroblasts in subepithelial connective tissue may play a role in maintaining epithelial integrity in health and in the apical migration of junctional epithelium in periodontitis. The aims of this study were (a) to compare expression levels of KGF and SF by periodontal ligament (PDL) and gingival fibroblasts; and (ii) to determine the effects of interleukin (IL)-1 beta, transforming growth factor (TGF)-beta 1, platelet-derived growth factor (PDGF)-BB and epidermal growth factor (EGF) on KGF/SF expression by these cell populations. Three paired PDL and gingival fibroblast strains were developed. The KGF and SF protein levels were analysed by enzyme-linked immunosorbent assay. Relative levels of KGF and SF mRNA in cytokine-treated cultures were determined using semiquantitative reverse transcriptase polymerase chain reaction. No differences in the levels of KGF and SF produced by PDL and gingival (SOG) populations were found. In both cell types IL-1 beta stimulated KGF and SF expression, while TGF-beta 1 significantly inhibited expression at both the mRNA and protein levels. Epidermal growth factor and PDGF-BB induced differing effects on expression, stimulating SF protein production but inhibiting KGF output in both fibroblast populations. Differences in response to EGF and PDGF were also seen between paired PDL and gingival fibroblasts. PMID- 12558808 TI - Immunocytochemical characterization of ectopic enamel deposits and cementicles in human teeth. AB - Despite the relative frequency and clinical relevance of radicular enamel deposits and cementicles, their etiology and nature are unknown. The purpose of the present study was therefore to evaluate the presence and distribution of mineralization-associated non-collagenous matrix proteins (NCPs) in various types of root-associated ectopic mineralizations. Human teeth were processed for embedding in epoxy or acrylic resins. Tissue sections were incubated with antibodies to amelogenins (AMEL), bone sialoprotein (BSP), and osteopontin (OPN). Radicular enamel deposits contained residual organic matrix that labeled for AMEL. In contrast, BSP and OPN were not detected in the residual enamel matrix, they were found in the cementum deposited on its surface as well as in collagen free cementicle-like structures in the adjacent periodontal ligament. True cementicles consisted of a collagenous matrix intermixed with a non-collagenous ground substance. Labeling for BSP and OPN was mainly associated with the interfibrillar ground substance. No immunoreactivity for AMEL was detected in cementicles. These data indicate that ectopic enamel deposits on the root retain a high amount of AMEL, whereas cementicles contain BSP and OPN, two NCPs typically found in bone and cementum. These NCPs may, like in their normal tissue counterparts, play a role in the mineralization process. PMID- 12558809 TI - Characterization of porcine dentin sialoprotein (DSP) and dentin sialophosphoprotein (DSPP) cDNA clones. AB - Dentin sialophosphoprotein (DSPP) is a chimeric glycoprotein with dentin sialoprotein (DSP) on its N-terminus and dentin phosphoprotein (DPP) on its C terminus. We have constructed and screened a unidirectional cDNA library derived from the pulp organ of developing pig teeth, and isolated cDNA clones encoding DSP-only, as well as two DSPP clones with alternative sequences in their 3' coding regions. The DSP-only transcript has an open reading frame of 386 codons, and is generated through the use of a polyadenylation signal within intron 4, immediately following the DSP coding region. the use of this polyadenylation signal deletes the DPP coding region and places a TGA translation termination signal as the fourth codon following the exon 4-encoded segment. The DSPP cDNAs contain open reading frames of 593 and 600 codons. Northern blots hybridized to radiolabeled DSP probes showed bands at 1.4, 2.5, 4.4, and 4.8 kb. Cloning and characterization of reverse transcriptase polymerase chain reaction products confirmed the existence of mRNA encoding pDSP386, pDSPP593, and pDSPP600in vivo, but also suggested that DNA sequence redundancies in the DSPP coding region make it prone to cloning artifacts. PMID- 12558810 TI - Effect of a combination of fluoride dentifrice and varnish on enamel surface rehardening and fluoride uptake in vitro. AB - The purpose of this study was to evaluate the effect of combining fluoride dentifrice (FD) and varnish (FV) on in vitro enamel surface rehardening and on fluoride uptake under a pH-cycling regimen. Seventy-eight bovine enamel blocks with early lesions were used and 52 were divided into four treatment groups: (a) placebo non-fluoridated dentifrice (PD); (b); FD (1100 p.p.m. F as NaF); (c); FV (Duraphat) + PD; and (d) FV + FD. The FV was applied to enamel blocks of groups FV + PD and FV + FD before the pH-cycling regimen, and all of them were submitted to dentifrice during cycling. Surface enamel microhardness was determined on the dental blocks before and after demineralization, and after the pH-cycling regimen. The percentage of surface microhardness recovery (%SMHR) was calculated. Fluoride in the blocks was also determined, after removing three layers of enamel. The highest values of percentage SMHR were observed for the FD group. The greatest fluoride uptake was found in the FD and FV + FD groups, but the difference between them was not statistically significant. It was found that the frequent use of fluoride dentifrice resulted in greater benefit in enamel surface rehardening, with a similar effect on fluoride uptake, when compared with its combination with a single fluoride varnish application. PMID- 12558811 TI - In vitro evaluation of marginal and internal adaptation after occlusal stressing of indirect class II composite restorations with different resinous bases. AB - Composite inlays are indicated for large cavities, which frequently extend cervically into dentin. The purpose of this study was to compare in vitro the marginal and internal adaptation of class II fine hybrid composite inlays (Herculite, Kerr) made with or without composite bases, having different physical properties. Freshly extracted human molars were used for this study. The base extended up to the cervical margins on both sides and was made from Revolution (Kerr), Tetric flow (Vivadent), Dyract (Detrey-Dentsply) or Prodigy (Kerr), respectively. Before, during and after mechanical loading (1 million cycles, with a force varying from 50 to 100 N), the proximal margins of the inlay were assessed by scanning electron microscopy. Experimental data were analysed using non-parametric tests. The final percentages of marginal tooth fracture varied from 30.7% (no base) to 37.6% (Dyract). In dentin, percentages of marginal opening varied from 9.2% (Tetric Flow) to 30.1% (Prodigy), however, without significant difference between base products. Mean values of opened internal interface with dentin varied from 11.06% (Tetric Flow) to 28.15% (Prodigy). The present results regarding dentin adaptation confirmed that the physical properties of a base can influence composite inlay adaptation and that the medium rigid flowable composite Tetric Flow is a potential material to displace, in a coronal position, proximal margins underneath composite inlays. PMID- 12558812 TI - Influence of food consistency on the rabbit masseter muscle fibres. AB - The plasticity of the masseter muscle was studied by comparing two groups of rabbits that were fed soft- and hard-diet for 87 d. Incisors of the soft-diet group were cut back to minimize the bite forces. Muscle fibres were immunohistochemically defined as fast- or slow-contracting fibres and their cross sectional area was measured. The muscles of animals fed with the hard-diet were composed of fibres with larger cross-sectional areas than the soft-diet group. The relative difference was larger in slow-contracting fibres than in fast contracting fibres. The results were similar for the different regions of the muscle. No changes in fibre composition were found. In conclusion, the difference in food consistency, as induced in this study, caused changes in the muscle fibre cross-sectional area that can be recognized from the altered necessary occlusal forces, which result from the modified forces developed by the masseter muscle. PMID- 12558813 TI - Steroid-sensitive nephrotic syndrome and vascular endothelial growth factor gene polymorphisms. AB - Genetic polymorphisms have been recognized as important determinants of gene expression. Three common single nucleotide polymorphisms have been identified in the promoter and 5' untranslated region of the vascular endothelial growth factor (VEGF) gene: -460 C --> T, -141 A --> C and +405 G --> C. As VEGF has been postulated to play a role in the pathogenesis of childhood steroid-sensitive nephrotic syndrome (SSNS), this study tested the hypothesis that VEGF genotype may be associated with susceptibility to SSNS. We examined the genotype frequencies of these polymorphisms in a total of 116 children with SSNS and 150 control subjects, using polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP). There were no statistically significant differences in any of the genotype frequencies between SSNS patients and controls. We conclude that VEGF -460, -141 and +405 genotypes are not associated with susceptibility to childhood SSNS. PMID- 12558814 TI - Genotyping for cytokine polymorphisms: allele frequencies in the Italian population. AB - It has been demonstrated that many cytokine genes [e.g. tumour necrosis factor alpha (TNF-alpha) and interleukin 10 (IL-10)] show polymorphisms which may affect gene transcription, causing individual variations in cytokine production. The majority of polymorphisms described are single nucleotide polymorphisms (SNPs). In 140 healthy Italian subjects, the allelic and genotype frequencies were determined for the cytokine genes IL-1 alpha (T/C -889), IL-1 alpha (C/T -511, T/C +3962), IL-12 (C/A -1188), interferon (IFN)-gamma (A/T UTR 5644), transforming growth factor (TGF)-alpha (C/T codon 10, G/C codon 25), TNF-alpha (G/A -308, G/A -238), IL-2 (T/G -330, G/T +166), IL-4 (T/G -1098, T/C -590, T/C 33), IL-6 (G/C -174, G/A nt565), IL-10 (G/A -1082, C/T -819, C/A -592), IL-1R (C/T pst11970), IL-1RA (T/C mspa111100) and IL-4RA (G/A +1902). All typings were performed with PCR-SSP assays. Allele and genotype frequencies and linkage disequilibria were calculated and compared with those of other populations. PMID- 12558815 TI - Complementary DNA sequence of the HLA-B*3924 allele. AB - We have isolated the complete coding region of HLA-B*39 from a Spanish Caucasoid, using a new PCR primer for its 5' untranslated region. The cDNA matched partial genomic sequences of B*3924, an allele whose distribution appears to be restricted to Mediterranean and Arabian Caucasoids. A single amino acid change exclusive to B*3924 (threonine-98) distinguishes it from B*3903. PMID- 12558816 TI - Human cytokine gene nucleotide sequence alignments: interleukin 4 receptor alpha chain (IL4RA), IL5RA, IL8, IL8RB and IL13. PMID- 12558817 TI - Nomenclature for factors of the HLA system, update July 2002. PMID- 12558818 TI - Nomenclature for factors of the HLA system, update August 2002. PMID- 12558819 TI - Nomenclature for factors of the HLA system, update September 2002. PMID- 12558820 TI - Nomenclature for factors of the HLA system, update October 2002. PMID- 12558823 TI - Research in epilepsy: development priorities for developing nations. AB - PURPOSE: To identify research priorities in epilepsy for developing nations. METHODS: A panel discussion with audience participation at the Indo-U.K. Workshop on Epilepsy. This included short presentations by panelists, the presentation of a research proposal, and debate on research priorities. RESULTS: The need to focus on primary-care populations; to use a multi-centre random block design; to incorporate rural areas and a service component; to study incidence, natural history, and aetiology; to focus on problems, such as cysticercosis, and to adopt a comprehensive public health-centred approach in doing so; to study disorders of local interest, such as hot water epilepsy; to pilot both pharmacological and nonpharmacological interventions; to incorporate comprehensive measures of cognition, behaviour, and psychosocial outcome in all studies; and to examine the role of novel diagnostic tools (imaging for example) and therapy (surgery for example) on cost were all outlined as priority areas. DISCUSSION: There is a felt need for greater and better-quality research output from the developing world. The development of uniform research protocols, the twinning of developed and developing nations for research, and training of developing nations' personnel are likely to increase research output in the years that come. PMID- 12558824 TI - Epidemiology of epilepsy in India. AB - The article briefly reviews literature pertinent to the incidence, prevalence, prognosis, etiology, and treatment gap of epilepsy in India. Despite defective methodology, Indian studies have shown prevalence rates comparable with those from other countries. No incidence studies exist. Prognosis for those with epilepsy is worse in developing countries. The role of infection, especially neurocysticercosis, in the etiology of epilepsy has still to be fully worked out, as must the relative contributions of the other known etiological factors. Neuroimaging is needed to calculate attributable risk. Reasons for the treatment gap are discussed briefly, pointing out that in India, people seek alternative forms of treatment concurrently. The occurrence and effects of stigma are mentioned. The article ends with the author's opinion on how and where the epidemiology of epilepsy should proceed in the future. PMID- 12558825 TI - Genetics in epilepsy: transcultural perspectives. AB - In the last two decades there have been many significant advances with regard to the genetics of human epilepsies. Knowledge of the mutations responsible for inherited human epilepsies not only will help in a better diagnosis and understanding of the mechanisms involved in epileptogenesis and their inheritance, but also will result in the development of better and safer antiepileptic drugs, discovery of novel therapeutic modalities, improved genetic counseling, and perhaps even prevention of epilepsies. The overall impact of these genetic discoveries across different populations and cultures will perhaps be beneficial to improving the lives of the millions affected by these diverse but common brain disorders. PMID- 12558826 TI - The natural history of epilepsy in the era of new antiepileptic drugs and surgical treatment. AB - PURPOSE: To review the factors influencing prognosis of epilepsy in this era of antiepileptic drugs and surgical treatment. METHODS: Systematic review of peer reviewed literature. RESULTS: The prognosis depends on many factors; aetiology, age of onset, number of seizures at onset, the natural history of the condition, and the influence of treatment. However, between 70 and 80% of people developing epilepsy will go into long-term remission, usually within the first 5 years. DISCUSSION: Prognosis in epilepsy is the prospect of attaining complete seizure freedom once a pattern of recurrent epileptic seizures has been established. Well designed prospective cohort studies using standard outcome measures need to be conducted, especially in non-Western cultures. PMID- 12558827 TI - Neuropsychiatric disorders in epilepsy: some transcultural issues. AB - PURPOSE: To review transcultural perspectives in the neuropsychiatry of epilepsy. METHODS: Systematic literature searches of standard databases, cross-referencing, chapters, and opinion leader articles. RESULTS: Articles from the Indian subcontinent, Africa, and Japan were identified and are reviewed herein. The spectrum of psychopathology in epilepsy is rather similar across cultures. However, psychopathology specific to epilepsy, the interictal behavioural syndrome of Geschwind, for example, has not been well studied outside the Western world. DISCUSSION: There is a need for well-designed epidemiological studies of neuropsychiatric disorders in epilepsy. These should use harmonised protocols and outcome measures. Special attention should be paid to the impact of aetiology on psychiatric co-morbidity and disablement. PMID- 12558828 TI - Solitary cerebral cysticercus granuloma. AB - Solitary cerebral cysticercus granuloma (SCCG) is one of the commonest causes of seizures in Indian patients. SCCG has been confused in the past with tuberculomas, but by applying a set of diagnostic criteria proposed by the author, they can be diagnosed accurately in the vast majority of patients. Patients with SCCG are managed effectively with antiepileptic drugs (AEDs). The role of cysticidal drugs in their management is controversial. SCCG resolves spontaneously at a variable rate and has a good seizure outcome, with >90% of patients remaining seizure free after discontinuation of AEDs. PMID- 12558829 TI - Hot-water epilepsy. AB - Geographically specific epilepsy syndromes are of epidemiologic interest and may assist in understanding the pathogenic mechanism of epilepsy in general. Some may have a genetic basis with added environmental influence. Among these epilepsy syndromes in India is an interesting type of reflex seizure known as hot-water epilepsy (HWE) and single small computed tomography (CT)-enhancing lesions (SSCTLs). PMID- 12558831 TI - Some problems and pitfalls in developing countries. AB - In developed countries with well-structured health care and reimbursement systems, epilepsy care is evidence based. By contrast, in developing countries, patients with epilepsy encounter several significant barriers to adequate treatment and are more often managed according to local ethnic, racial, religious, economic, educational, and cultural diversities. Cost is one issue that clearly determines antiepileptic drug (AED) selection, and it is reasonable to recommend one of the traditional, and cheaper, AEDs as first-line therapy. However, the appropriate choice of drug in an individual patient is a balance of efficacy, tolerability, and cost and should be tailored to individual affordability. PMID- 12558830 TI - Clues to the genetic influences of drug responsiveness in epilepsy. AB - Despite the introduction of newer antiepileptic drugs (AEDs), nonresponsiveness to AEDs remains a common problem in epilepsy clinics. There may be important genetic determinants for responsiveness, and this discussion focused on some potential areas: drug transporters; drug-metabolizing enzymes, and ion channels. We review the literature and speculate the contribution of each of these factors in management of patients with epilepsy in the future. PMID- 12558832 TI - Current status of surgery in the management of epilepsy. AB - PURPOSE: To review systematically the available evidence with regard to the current status of epilepsy surgery in the management of patients with epilepsy. METHODS: A careful search of published literature, including Medline, published reviews, chapters, and cross-references thereof. RESULTS: With medical treatment of epilepsy being unsuccessful in many cases, the importance of surgical approaches cannot be underscored. Early surgery is the treatment of choice for patients with clear-cut mesial temporal sclerosis and results in significant clinical improvement in up to 80% of cases, provided the EEG, neuropsychological, and neuropsychiatric results are in concordance with this approach. In patients with poorly defined, widespread, or dual pathology, however, invasive recordings may be necessary, and while this is performed in major centres, the outcome is rather more variable in this group. Improved surgical techniques, and the use of stereotactic approaches and image guidance procedures, have resulted in surgical resections becoming more selective. With isolated structural lesions such as dysembryoplastic tumours, low-grade astrocytomas, or focal vascular abnormalities, total macroscopic and radiological evidence of lesional excision is associated with excellent seizure-free outcome. The first randomised controlled trial of epilepsy surgery has demonstrated clearly the efficacy of these techniques, and the risk of complications. DISCUSSION: Increasing sophistication of noninvasive presurgical evaluation enables surgical candidates to be identified at an earlier stage and presents a realistic alternative to medical treatment in many cases. The introduction of minimally invasive techniques has had a significant impact on surgical practice and its associated morbidity. The future of epilepsy surgery lies with continued basic science research and its application to clinical medicine. PMID- 12558833 TI - Problems and pitfalls in developing countries. AB - Despite major challenges, in the last decade several epilepsy centers in the developing world have successfully implemented epilepsy surgery programs and produced results comparable to those from developed countries. If the program is to have a lasting impact, it would be essential to work with and educate the local professionals and public about the recent advances in the treatment of epilepsies. The epilepsy surgery centers in developing countries should initially restrict their surgical candidates to patients with mesial temporal lobe epilepsy and those with circumscribed potentially epileptogenic lesions in whom the epileptogenic zone can be unquestionably localized by using locally available relatively inexpensive and noninvasive technologies, and in whom an excellent postoperative outcome can be guaranteed. It is important for such epilepsy center to assess its capabilities and limitations regularly and adopt a stepwise progressive approach to increasing levels of complex presurgical evaluation and surgical treatment strategies. PMID- 12558834 TI - Perspectives from a developed nation. AB - Epilepsy is the most common serious neurologic disorder, affecting 350,000 people in the United Kingdom. There are five neurologists per 1,000,000 population, which is better than in India but much lower than in other developed nations. Thus, a patient's day-to-day prescribing, supervision, and support depends on primary care. In the U.K., patients are entitled to register with a general practitioner (GP), who has an average of 1,841 patients. Seventy-eight percent of patients will consult their GP annually. Patients in the U.K. find that GPs are accessible and have good communication skills. There is, however, inadequate time in short consultations to provide the quality of care suggested by the latest review of services for patients with epilepsy (CSAG). This is further complicated by the heterogeneous and stigmatising nature of the condition. An improvement in the process of care in the primary-care setting can result from three important strategies: appropriately trained practice nurses running practice nurse-led clinics; structured management of care, possible because of the unique system of registration, which facilitates audit, prescription monitoring, and recall; and, finally, improved teamwork and communication based on protocols locally agreed upon between primary and secondary care. The future will tell whether these initiatives will improve the outcomes of care. PMID- 12558835 TI - Perspectives from a developing nation with special reference to rural areas. PMID- 12558836 TI - Epilepsy control program in India: a district model. AB - Based on data originating from neuroepidemiologic studies in India, epilepsy has been identified as a public health problem. The treatment gap varies from 50 to 70% among persons with epilepsy and is compounded further by limited availability of neurologists, often working in urban metropolises, thus restricting the availability of services in a predominantly rural agrarian country. Cost effective, sustainable epilepsy care programs are urgently required in India. To bridge this wide gap, a district model was developed with the central focus on training the district medical officers in providing epilepsy care with sensitization of state health administrators and nodal neurologists to support and sustain the program. Workshops were conducted for all three groups during the period 1999-2001, with the involvement of 21 state health administrators, 148 district medical officers, and 28 neurologists. Preliminary evaluation indicates a growing interest, enthusiastic commitment, and willingness among health professionals to expand epilepsy care to the periphery. The program revealed that it is possible to reach people with epilepsy on the premise of early diagnosis and management, need-based referral network, and a rational planning for uninterrupted availability of antiepileptic drugs along with education of families. Policy-level initiatives are required further to transform this on a wider scale and to evaluate the efficacy and effectiveness of this approach. PMID- 12558837 TI - A study of perceptions of facial hemangiomas in professionals involved in child abuse surveillance. AB - Facial hemangiomas are common benign tumors of infancy. They can mimic the appearance of bruises and parents report false accusations of child abuse by strangers. We investigated perceptions of facial hemangiomas in two professional groups involved in child abuse surveillance. Thirty health visitors and 30 primary school teachers were sent a clinical case questionnaire about a child with an involuting facial hemangioma with a color photograph of the lesion. Nineteen health visitors (63%) and 18 primary school teachers (60%) responded. Seventy-four percent of the health visitors and 11% of the teachers correctly diagnosed a hemangioma. Fifty percent of the teachers were unable to decide on a diagnosis. However, only one health visitor (5%) and one teacher (5%) thought that the lesion was a nonaccidental injury and would involve a child protection advisor. About one-third of health visitors and one-third of teachers expressed concerns that the child could suffer psychologically from teasing by peers at school and about one-third of health visitors were also concerned about the psychological effects of the lesion on the child's parents. Health visitors and teachers were most unlikely to mistake the facial hemangioma in this study for a nonaccidental injury. PMID- 12558838 TI - The family impact of atopic dermatitis in children: the role of the parent caregiver. AB - Although some preliminary work exists examining the impact of atopic dermatitis (AD) in children on their families, there is no empirical work examining specific parent caregiver factors that could contribute to the family impact of this condition. We conducted a cross-sectional, exploratory analysis of how parent caregivers are affected by their child's AD, and how certain parent caregiver characteristics and perceptions affect the family impact of this condition. Parent caregivers of children with AD (n = 49) were administered a survey to collect detailed data on socioeconomic status, health perceptions, and caregiving issues. Family impact of the child's AD was measured using a modified AD Family Impact Scale. Multiple regression analyses revealed that three major factors associated with the parent caregiver were correlated with large increases in the family impact scores: 1) perception that the child's condition is severe (13%, p < 0.01), 2) high use of nonmedical services for child's condition (21%, p < 0.01), and 3) financial concern about the child's condition (18%, p < 0.01). These preliminary data indicate distinct characteristics of the parent caregiver that are associated with higher family impact of AD in children. These parent caregiver factors may be important in identifying suitable audiences and areas for education for optimal management of children's AD. PMID- 12558839 TI - Treatment of psoriasis in children: is there a role for antibiotic therapy and tonsillectomy? AB - Numerous studies implicate subclinical or recurrent streptococcal infection as a trigger or maintenance factor in the pathogenesis of psoriasis in children. The purpose of this article is to review the efficacy of antibiotic therapy and tonsillectomy as treatments for childhood psoriasis. Clinical trials assessing the efficacy of antibiotics or tonsillectomy as treatments for childhood psoriasis were identified with a search of the medical literature and the results were compared. Only one controlled clinical trial was identified and it did not find a significant effect of antibiotic treatment on psoriasis. In other studies, the percentage of psoriasis patients who experienced disease clearance with antibiotic therapy ranged from 0% to 55%, with no patients experiencing disease worsening during treatment. No controlled trials of tonsillectomy for psoriasis were identified. The percentage of patients who experienced disease clearance after tonsillectomy in uncontrolled trials ranged from 32% to 53% and a similar percentage reported significant improvement in their psoriasis, with a maximum of 7% noting worsening of the disease after the operation. The available evidence does not demonstrate the efficacy of either antibiotic therapy or tonsillectomy in the treatment of childhood psoriasis. Because these treatments are relatively benign compared to other treatments for severe psoriasis, the use of antibiotic therapy or tonsillectomy may still be worth considering, especially for those patients with recurrent streptococcal infections that seem to trigger or maintain their skin disease. PMID- 12558840 TI - Skin cancer prevention counseling and clinical practices of pediatricians. AB - Little is known about pediatricians' counseling and clinical practices to reduce skin cancer risk among their patients. Thus our objectives were to characterize skin cancer preventive counseling and clinical practices in a sample of pediatricians and identify correlates of these practices. Physicians practicing general pediatrics in Harris County, Texas, received a mail survey that assessed their sun protection recommendations and skin cancer preventive counseling and clinical practices. Pediatrician, patient, and medical practice variables were assessed as correlates. Most (76%) pediatricians routinely recommended sunscreen; however, relatively few (24%) suggested reapplying it after prolonged periods outside. About half routinely recommended protective clothing (53%), shade (47%), or limiting midday sun exposure (46%). Even fewer pediatricians routinely discussed skin cancer risk factors, passed out sunscreen samples, made educational materials available, took a family history of skin cancer, or documented risk factors in a patient's chart. More than half reported that they routinely performed full-body skin examinations during a first visit (65%) and annually (56%). Perceived barriers, perceived relevance of skin cancer prevention, and personal sun protection practices were important factors associated with professional practices in this sample. Interventions are needed to increase pediatricians' counseling and clinical practices to reduce skin cancer risk among patients. PMID- 12558841 TI - Neu-Laxova syndrome: a case report. AB - Neu-Laxova syndrome is a rare congenital disorder characterized by microcephaly, limb contactures, lissencephaly and ichthyosis. A case of Neu-Laxova syndrome is presented, with a discussion of clinical manifestations, complications, and therapeutic interventions. PMID- 12558842 TI - Rosacea-like demodicidosis in an immunocompromised child. AB - Demodex folliculorum is a saprophytic mite of the human pilosebaceous unit. It is rarely found in children. It has been implicated in the development of follicular pityriasis, rosacea-like demodicidosis, pustular folliculitis, blepharitis, and granulomatous rosacea. We describe a 4-year-old boy who developed asymptomatic facial lesions that histologically corresponded to demodicidosis. He was in clinical remission of acute lymphoblastic leukemia and currently receiving maintenance chemotherapy. Exanthems related to D. folliculorum are rare in children. Most cases occur in immunocompromised patients and the clinical and histologic findings are diverse. A differential diagnosis should be established with rosacea and perioral dermatitis. The role of Demodex in the pathogenesis of these disorders is controversial. Immunosuppression might increase the number of mites, favoring an inflammatory reaction, or there could exist an impaired cutaneous immunologic response to the parasites. PMID- 12558843 TI - Subacute cutaneous lupus erythematosus in childhood. AB - We report a 7-year-old girl who presented with erythematous-infiltrated, figurate, well-defined lesions over sun-exposed skin and antinuclear and Ro/SSA antibodies. Lupus band test revealed granular IgM and microgranular C3 deposits at the dermoepidermal junction. Histopathologic examination of lesional skin showed orthohyperkeratosis, epidermal atrophy, widespread hydropic degeneration of the epidermal basal cell layer, and a dermal perivascular lymphohistiocytic infiltrate. The clinical, immunopathologic, and histopathologic findings were consistent with the diagnosis of annular-polycyclic variant of subacute cutaneous lupus erythematosus (SCLE). Treatment with hydroxychloroquine cleared the cutaneous lesions. We report this patient because SCLE is extremely rare in childhood, and discuss the two previous cases reported in the literature. PMID- 12558844 TI - Self-healing juvenile cutaneous mucinosis. AB - A healthy 14-month-old black girl presented with a 3-week complaint of "knots" on the face and hands. The lesions were acute in onset and asymptomatic. Multiple, firm, nontender, skin-colored to erythematous nodules were noted on the scalp, forehead, axillae, lower legs, abdomen, and hands. A skin biopsy specimen revealed a well-circumscribed accumulation of mucin in the reticular dermis. Colloidal iron stain was positive. Radiographs showed soft tissue prominence only. Serum protein electrophoresis, thyroid function tests, complete blood count, sedimentation rate, and antinuclear antibody were normal, except for lymphocytosis. Findings were consistent with self-healing juvenile cutaneous mucinosis (SHJCM). SHJCM is a condition of unknown etiology characterized by rapid onset of asymptomatic, indurated papules or nodules. Affected children may have arthralgias, but are otherwise well. Spontaneous resolution is the rule. Most skin lesions in our patient had resolved within 6 months of onset. This patient is unique because of the young age of onset. PMID- 12558845 TI - Cutaneous silica granuloma in a child. AB - A 12-year-old girl had a 4-year history of two asymptomatic, sharply demarcated, granulomatous lesions on her face. The lesions did not respond to topical steroids and histopathologic examination of biopsy specimens showed granulomatous inflammation. Since cutaneous leishmaniasis is endemic where the patient lived, she was diagnosed as chronic cutaneous leishmaniasis but did not respond to meglumine antimoniate treatment. Reexamination of the biopsy specimens under polarized light revealed numerous birefringent crystalline particles, and cutaneous silica granuloma was the final diagnosis. The lesions were treated with intralesional triamcinolone acetonide and completely disappeared. We report this case of cutaneous silica granuloma, which is unusual in children, and emphasize the importance of polarized light microscopic examination of granulomatous skin diseases. PMID- 12558846 TI - Verruciform xanthoma in association with milroy disease and leaky capillary syndrome. AB - An 18-year-old Caucasian boy with Milroy disease involving the right leg presented with erythematous, smooth-topped, waxy papules on the dorsum of his right foot. A 12-year-old Caucasian girl with leaky capillary syndrome presented with hemorrhagic verrucous papules on the dorsum of the toes of both feet. Histopathologic analysis revealed changes consistent with xanthoma. Both patients were treated with leg compression, curettage, and electrodesiccation. Although similar papules have been described in the setting of lymphedema from other causes, this is the first report of verruciform xanthoma associated with Milroy disease or leaky capillary syndrome. PMID- 12558847 TI - Ichthyosis follicularis: a case report and review of the literature. AB - Ichthyosis follicularis (IF) is a very rare neurocutaneous, X-linked recessive condition affecting the skin, hair, eyes, and central nervous system (CNS). This report describes a child with facial dysmorphism, mental retardation, psychomotor delay, congenital alopecia of the scalp, eyebrows, and eyelashes, and extensive spiny follicular papules. A skin biopsy specimen showed the characteristic absence of sebaceous glands. We also reviewed the literature on this very rare entity. Additional findings observed in our patient, including hepatosplenomegaly, undescended testicles, and ptosis, have not been reported before. PMID- 12558848 TI - Stevens-Johnson syndrome: a diagnostic challenge in the absence of skin lesions. AB - We report a 14-year-old boy with Stevens-Johnson syndrome who presented with mucous membrane lesions but without skin lesions and discuss the differential diagnosis of oral mucous membrane lesions in childhood. Stevens-Johnson syndrome in children is most frequently caused by a Mycoplasma pneumoniae infection. The full clinical picture of Stevens-Johnson syndrome can be present before seroconversion of Mycoplasma antibodies is observed. One should keep in mind that one negative titer of Mycoplasma antibodies does not rule out M. pneumoniae infection. PMID- 12558849 TI - Juvenile subcorneal pustular dermatosis: a case report. AB - Subcorneal pustular dermatosis is a chronic, relapsing, pustular dermatosis that usually develops in elderly women. It is rarely seen in childhood and adolescence. The hallmark of the disease is formation of a subcorneal pustule detected on histopathologic examination. Herein we present a 13-year-old girl diagnosed with subcorneal pustular dermatosis based on the typical clinical and histologic findings. Direct and indirect immunofluorescence and serum protein electrophoresis had negative results. The patient partially recovered after 5 weeks of treatment consisting of oral dapsone and a topical steroid ointment. PMID- 12558850 TI - Central nervous system involvement in neonatal lupus erythematosus. AB - Computerized tomography (CT) of the brain was performed in 10 of 11 consecutive infants with neonatal lupus erythematosus (NLE) (five boys and six girls). Ten of the 11 infants had brain neurosonography. Nine of 10 infants had abnormal CT scans. There was diffuse, markedly reduced attenuation of the cerebral white matter in four infants studied in the first week of life, and also in an infant 5 weeks of age. Patchy reduced subcortical white matter attenuation was observed in another 5-week-old infant. Basal ganglia calcifications were present in two infants at 2 months of age, one of whom also had mild ventriculomegaly. A patient with macrocephaly studied at 4 months of age had enlarged ventricles and subarachnoid spaces consistent with benign macrocephaly of infancy. Cerebral ultrasound examination was abnormal in all five infants studied in the first week of life and in one infant at 2 months of age. Findings included subependymal cysts (4), echogenic white matter (3), and echogenic lenticulostriate vessels (3). Apart from one case of macrocephaly, there was no clinical evidence of neurologic disease and the subsequent development of these infants has been normal. Subclinical central nervous system (CNS) disease in NLE is likely to be a transient phenomenon that resolves as maternal antibodies are cleared from the infant's circulation. It is important to be aware of these neuroimaging abnormalities to avoid misdiagnosis of congenital viral infection in a newborn with multisystem NLE. The potential for neurologic sequelae is uncertain. PMID- 12558851 TI - A case of infantile acne due to pityrosporum. AB - We report a case of infantile acne. The lesion on the forehead resembled infantile acne clinically, but histologically it contained yeasts of Pityrosporum. The lesion was treated with topical ketoconazole cream and improved. We suggest that Pityrosporum is associated with infantile acne. PMID- 12558852 TI - Phototherapy in pediatric patients. AB - The treatment of children with psoriasis, atopic dermatitis (AD), pityriasis lichenoides, and scleroderma poses a therapeutic problem because all therapeutic options are associated with numerous side effects. Therefore ultraviolet A and B (UVA and UVB) phototherapy is presented as a possible alternative to some of these therapies, primarily topical and systemic corticosteroids, in children. Our results in treating children with phototherapy and psoralen plus UVA (PUVA) bath phototherapy over the past 5 years are reported. UVB therapy (TL01) was used in 20 psoriatic children (6 boys, 14 girls; ages 6-14 years) during the stage of disease exacerbation and in 9 children (3 boys, 6 girls; ages 8-16 years) with pityriasis lichenoides. Combined UVA/UVB phototherapy was applied in 21 AD children (7 boys, 14 girls; ages 4-15 years). Photochemotherapy with local application of a PUVA bath was used in six children (2 boys, 4 girls; ages 9-16 years) with circumscribed scleroderma and in one girl with systemic scleroderma. All children received short courses of phototherapy with either no maintenance or short maintenance. All three therapeutic protocols resulted in a certain degree of improvement in most of the study patients. None of the patients exhibited any early phototherapy side effects. We conclude that phototherapy and PUVA bath are valuable and safe therapeutic options for selected children who do not respond to other treatments. PMID- 12558853 TI - What syndrome is this? Neu-Laxova syndrome. PMID- 12558854 TI - Crusted red papules on the head and trunk. PMID- 12558855 TI - Wart therapy reconsidered. PMID- 12558856 TI - Diagnostic difficulties in lymphedema distichiasis. PMID- 12558859 TI - Skin disorders in young travelers recently returned from overseas destinations: a French multicentric prospective study. PMID- 12558860 TI - Tuberculid in a child: transformation from papulonecrotic to lichen scrofulosorum. PMID- 12558858 TI - Treatment of granuloma annulare with topical 5% imiquimod cream. PMID- 12558861 TI - Anthrax of the eyelid in a Turkish girl. PMID- 12558863 TI - Lobular capillary hemangioma of the oral mucosa: clinicopathological study of 43 cases with a special reference to immunohistochemical characterization of the vascular elements. AB - Clinical and histopathological features were investigated in 43 cases of oral lobular capillary hemangiomas (LCH) with a special reference to characteristics of the vascular elements. The lesions affected females more than males by a ratio of 1:1.5. Average age of the patients was 52.7 years. The lesions involved the gingiva (n = 15), the tongue (n = 13), the labial mucosa (n = 10) and other sites. The lesions appeared usually as a pedunculated mass with ulceration; size of the lesions was up to 15 mm. Histologically, a lobular area and an ulcerative area were distinguished. The density of vessels was about 1045/mm2 and 160/mm2 in the lobular and ulcerative areas, respectively. The average diameter of the vascular lumen was 9.1 5.6 mm (range: 2.8-42.0 mm) and 18.8 20.9 mm (range: 5.6 139.7 mm) in the lobular and ulcerative areas, respectively. In the lobular area, most of the vessels had an inner layer of endothelial cells showing positive reaction for von Willebrand factor (vWF) and CD34, as well as an outer layer of mesenchymal cells showing positive reaction for alpha-smooth muscle actin (ASMA). However, in the ulcerative area, there was a variety of types of vessels consisting of various proportions of both endothelial and ASMA-positive perivascular mesenchymal cells. These results indicate that most of the vascular elements in the lobular area resemble more pericapillary microvascular segments than they do capillaries. Thus, the authors propose the term 'lobular pericapillary hemangioma' to represent this type of lesion. PMID- 12558864 TI - Osteopontin is involved in the formation of epithelioid granuloma and bile duct injury in primary biliary cirrhosis. AB - Recently, it was shown that osteopontin (OPN) is involved as a chemoattractant cytokine in the recruitment of macrophages and T lymphocytes in the granulomas of diverse etiologies and also plays an important role in the production of autoantibodies and development of autoimmune diseases. Primary biliary cirrhosis (PBC) is characterized by immune-mediated bile duct damage with frequent epithelioid granulomas. In this study, the expression of OPN was immunohistochemically examined in 25 PBC and 52 control livers. Epithelioid cells within granuloma in PBC expressed OPN variably. These cells were also positive for CD68, suggesting their histiocyte/macrophage lineage. In addition, strong expression of OPN was seen in the cytoplasm of mononuclear cells infiltrating around granulomas and also damaged bile ducts in PBC. The number of such positive mononuclear cells and the ratio of OPN-positive cells/total infiltrating cells in portal tracts were higher in PBC than in controls. The majority of these OPN positive cells were found to be IgG- or IgM-producing plasma cells. These suggest that in PBC, OPN is an important immune molecule in portal tracts, and contributes to the recruitment of mononuclear cells into epithelioid granuloma and also participates in bile duct injury via B-cell differentiation and plasma cell expansion. PMID- 12558865 TI - Smad expression of hepatic stellate cells in liver cirrhosis in vivo and hepatic stellate cell line in vitro. AB - Smad expressions, signaling mediators of transforming growth factor-beta (TGF beta) superfamily of cytokines, were investigated in paraffin-embedded tissue sections of liver cirrhosis due to the hepatitis C virus infection and in the hepatic stellate cell (HSC) line in vitro. Smad 2/3, 4 and 7 was expressed in the nucleus of the HSC in the cirrhotic liver, while the expression was weak in the non-cirrhotic liver. TGF-beta1 expression in the HSC of the cirrhotic liver was strong, while the expression was weak in the non-cirrhotic liver. In situ hybridization also demonstrated the Smad signalings in the HSC of the cirrhotic liver, which confirmed the results of the Smad expressions by immunohistochemistry. The HSC line showed a cytoplasmic and a weak nuclear expression of Smads without TGF-beta1 stimulation, while these cells showed a strong Smad expression in the nucleus by TGF-beta1 stimulation. Immunocytochemical assay demonstrated that the TGF-beta1 stimulation induced the increase of the Smad expressions and the decrease of the autocrine TGF-beta1 in the HSC line. In situ hybridization assay also demonstrated an increase of the Smad mRNA signalings by TGF-beta1 stimulation in vitro. These observations suggest that the Smad expressions increase in the nucleus of the HSC in the cirrhotic liver and that the TGF-beta1 stimulation induces the Smad expression. PMID- 12558866 TI - Application of the AMeX method to the evaluation of HER-2 status in breast carcinomas: comparison with the results of HercepTest. AB - The recent development of the recombinant humanized monoclonal antibody against HER-2 oncoprotein requires a simple and accurate method for the evaluation of HER 2 status in patients with breast cancers. We here report that the evaluation of the HER-2 status is improved by the use of the acetone-methanol-xylene (AMeX) method. Compared with an ordinary test of HercepTest, 25 out of 63 cases (39.7%) were scored upwards by the AMeX method. In addition, the HER-2 gene amplification was easily estimated by fluorescence in situ hybridization (FISH) using the AMeX method. Thus, the AMeX method is likely to provide more improved data about the HER-2 status in breast carcinoma specimens. PMID- 12558867 TI - Pleomorphic adenoma with extensive adenoid cystic carcinoma-like cribriform areas of parotid gland. AB - We report an interesting case of pleomorphic adenoma (PA) with extensive adenoid cystic carcinoma (ACC)-like cribriform areas of parotid gland along with a discussion of the differential diagnosis. The patient was a 62-year-old-female who presented with a 5-month history of a slow-growing painless mass in the left preauricular region. Pathological examination of the excised mass revealed a well encapsulated tumor consisting of typical PA areas and extensive ACC-like cribriform areas. The transition between both areas was frequently observed. There was no clinical or histological evidence of a longstanding PA, such as prominent hyalinization with dystrophic calcification. Immunohistochemically, the neoplastic myoepithelial cells of the PA areas showed frequent expression of vimentin and S-100 protein, along with occasional positivity for glial fibrillary acidic protein and alpha-smooth muscle actin. The immunoreactivity of the neoplastic myoepithelial/basal cells forming cribriform nests was essentially identical to that of the PA areas. The difference of immunoreactivity of the basal/myoepithelial cells between PA and ACC may have some discriminatory advantage. In addition, the low proliferative activity observed in the entire tumor suggested that the cribriform nests resembling ACC are also the component of PA. PMID- 12558868 TI - Mixed tumor of deep soft tissue. AB - Mixed tumors of the salivary gland and skin are relatively common but are quite rare in soft tissue. It is believed that, as in the salivary gland form, most of these lesions are benign, but that a small subset behave in an aggressive fashion. We report here a patient with recurrent mixed tumor of soft tissue with infiltrative growth. The primary tumor arose in deep subfascial soft tissue of the right lower leg and was adjacent to the surface of the fibula. An open biopsy and complete resection were performed. Upon histological examination of the resected specimen, neoplastic cell infiltration at the tumor/soft tissue interface was not obvious; local recurrence, however, was observed 1 year later. The patient was treated with wide resection. Histological examination confirmed that the recurrent tumor with an extensive chondroid area invaded the osseous tissue of the fibula. At present, 1 year after the second resection surgery, there is no evidence of disease. PMID- 12558869 TI - Solitary fibrous tumor with malignant potential arising in sublingual gland. AB - A rare case is described of a solitary fibrous tumor (SFT) with malignant potential arising in the sublingual gland. A 59-year-old man presented with a 4 month history of a slowly enlarging painless mass in the center of the floor of the mouth. The tumor was a well-demarcated, firm mass with a multicystic lesion. The tumor exhibited highly cellular areas of spindle cells with patternless architecture alternating with hypocellular areas. The tumor cells were positive for CD34 and bcl-2 as well as vimentin, and negative for epithelial, myogenic, neurogenic and histiocytic markers. The tumor cells formed multiple satellite nodules around dilated ducts in the multicystic lesion, indicating infiltrative growth. In addition, areas exhibiting higher cellularity with increased mitoses were noticed in the satellite nodules, although cellular atypia was not obvious. These findings led to a final diagnosis of SFT with malignant potential. There has been no recurrence or metastasis for 27 months after the surgery. Solitary fibrous tumor of the salivary gland must be differentiated from various spindle cell neoplasms including myogenic, peripheral nerve sheath, fibroblastic and fibro-histiocytic spindle cell neoplasms, hemangiopericytoma and myoepithelioma. In addition to characteristic morphological features, an immunohistochemical positivity for CD34 and bcl-2 may aid in the diagnosis of SFT. PMID- 12558870 TI - Case of postradiation osteosarcoma with a short latency period of 3 years. AB - Osteosarcoma is one of the neoplasms that may occur following exposure to radiation. A case of osteosarcoma arising in the craniofacial bone with a short latency period of 3 years after radiotherapy for maxillary squamous cell carcinoma is described. A 64-year-old-man underwent right partial maxillectomy and chemoradiotherapy due to squamous cell carcinoma of his right maxillary sinus. Histologically, the tumor was composed of moderately differentiated squamous cell carcinoma with a component of spindle cell carcinoma. Three years later, osteosarcoma developed in the craniofacial bone within the irradiation field of the first tumor. Detailed histological examination demonstrated that there was no component of osteosarcoma in the first tumor or squamous cell carcinoma in the second tumor. Radiation-induced osteosarcoma usually occurs after a long latency period of more than 10 years after the radiotherapy. In this case, osteosarcoma was possibly a radiation-induced osteosarcoma with a short latency period of 3 years. PMID- 12558871 TI - Tumorous deformity of mitral valve leaflet after chordal rupture in a child. AB - A case with tumorous deformity of the posterior mitral valve leaflet after spontaneous chordal rupture in a child is described. A partial rupture in the chordae tendineae of the posterior mitral leaflet was found by echocardiography in a 9-year-old Japanese boy. Tumorous bulging was gradually developed in the leaflet and was surgically excised 5 years later. Multiple nodular tumors were found on the atrial surface of the posterior mitral leaflet. Histological examination revealed that the tumorous bulging consisted of myxomatous materials in which collagen fibrils and very fine elastic fibers were distributed loosely and irregularly. Normal-looking endothelial cells covered the luminal surface of the bulging lesion. Vimentin-positive spindle-shaped mesenchymal cells were scattered in the bulge area. The labeling index of proliferating cell nuclear antigen (PCNA) in these cells was 29.3%. These spindle cells were positive for matrix metalloproteinase (MMP)-1 in the entire bulge area. The cells and matrix were positive for MMP-2 and tissue inhibitor of MMP (TIMP)-1 in the basal area of bulging, but were weakly positive or negative at the surface area. Reactivity for TIMP-2 in the cells in the bulge area was obviously weaker than that in the cells at the spongiosa of the anterior mitral leaflet, which was obtained from the patient at the valve replacement operation 9 months after the initial operation. These findings indicated that the tumorous deformity of the mitral valve was formed by the overgrowth of valve tissue under the stimulation of mitral regurgitation in this child, and the imbalance of MMP and TIMP might play an important role in the bulge formation. PMID- 12558872 TI - Colliding primary lung cancers of adenosquamous carcinoma and large cell neuroendocrine carcinoma. AB - We report an extremely rare case of primary lung cancer showing various histological elements diagnosed as the collision of an adenosquamous carcinoma and a large cell neuroendocrine carcinoma by loss of heterozygosity (LOH) analysis of the human androgen receptor (AR) and phosphoglycerate kinase (PGK-1) genes. The tumor exhibited a tiny ground-glass opaque shadow suggesting atypical adenomatous hyperplasia 18 months prior to surgery. However, the tumor grew rapidly, and the resected tumor consisted of two closely located nodules. The larger nodule was composed of well-differentiated adenocarcinomatous and moderately to poorly differentiated squamous cell carcinomatous elements, while the smaller nodule consisted of a large cell neuroendocrine carcinomatous element with partial squamoid differentiation having focal continuity with the adenocarcinomatous element. Both the adenocarcinomatous and squamous cell carcinomatous elements revealed transitional features and LOH of AR and PGK-1 genes, while the large cell neuroendocrine carcinomatous element showed a monoclonal pattern but possessed both alleles of AR and PGK-1 genes. From these clinical and pathological results, the parental cell of the large cell neuroendocrine carcinomatous element was considered to be different from that of the adenosquamous carcinomatous element. PMID- 12558874 TI - 'Full of mercy and good fruits'. PMID- 12558875 TI - Observation during early clinical exposure - an effective instructional tool or a bore? PMID- 12558876 TI - Clinical supervision of SpRs: where does it happen, when does it happen and is it effective? PMID- 12558877 TI - Are orals worth talking about? PMID- 12558878 TI - Teaching sexual history taking to medical students and examining it: experience in one medical school and a national survey. AB - OBJECTIVES: To assess the feasibility and acceptability of training and examining medical students in taking a sexual history and to compare practice with other medical schools in the UK. DESIGN: A training programme involving group work, role play and clinical attachments was developed and applied to 131 students at the University of Bristol Medical School. They then underwent an objective structured clinical assessment using simulated patients. The practice of other medical schools was surveyed by postal questionnaire. RESULT: The students felt that the examination was a good test of their knowledge and skills. One student failed. Sexual history taking is taught in 17 of 22 medical schools but examined in only six. CONCLUSION: Both teaching and examining of sexual history taking skills are possible and are likely to occur increasingly in UK medical schools. PMID- 12558879 TI - An analysis of standardised patient checklist errors and their effect on student scores. AB - BACKGROUND: The frequency and nature of standardised patient (SP) recording errors during clinical performance examinations (CPX) have an effect on case scores and ultimately on pass/fail decisions. PURPOSE: To determine the effect of SP recording errors on case scores. METHODS: Standardised patients completed checklists immediately after each encounter. To determine checklist accuracy, multiple reviewers developed a checklist key for each student encounter studied. The total errors, the net errors, the errors of commission and omission and error rates by competency skill were analysed. RESULTS: The frequency of errors in history taking was greater than in physical examination, and the majority of errors were made in the students' favour. Summing the errors of commission and omission decreased the effect of total errors on student scores. CONCLUSIONS: High levels of SP recording accuracy are achievable. When errors occur, the net effect is usually in the students' favour. PMID- 12558880 TI - The rating of pre-clerkship examination questions by postgraduate medical students: an assessment of quality and relevancy to medical practice. AB - OBJECTIVE: To determine whether postgraduate students are able to assess the quality of undergraduate medical examinations and to establish whether faculty can use their results to troubleshoot the curriculum in terms of its content and evaluation. SUBJECTS: First and second year family medicine postgraduate students. MATERIALS: A randomly generated sample of undergraduate medical examination questions. METHODS: Postgraduate students were given two undergraduate examinations which included questions with an item difficulty (ID) > 0.60. The students answered and then rated each question on a scale of 1-7. RESULTS: The percentage of postgraduate students answering each question correctly correlated significantly with the average perceived relevance (Examination 1: r=0.372; P < 0.05; Examination 2: r=0.458; P < 0.05). Questions plotted for average postgraduate/undergraduate performance ratio versus the average perceived relevance were significantly correlated (Examination 1: r=0.462; P < 0.01; Examination 2: r=0.458; P < 0.05). CONCLUSIONS: This study offers a method of validating question appropriateness prior to examination administration. The design has the potential to be used as a model for determining the relevancy of a medical curriculum. PMID- 12558881 TI - What do students want? The types of learning activities preferred by final year medical students. AB - CONTEXT: A well-aligned curriculum has consistent focus on curricular goals, teaching/learning activities and assessment. Poor alignment affects the way students budget learning time and may interfere with achievement of curricular goals. We noticed students' poor attendance in clinical clerkships prior to final examinations and hypothesised that they were responding to curricular misalignment. OBJECTIVES: To quantify the extent to which students and tutors perceived a mismatch between activities pertaining to their current clerkship and those they thought necessary to prepare for final examinations and internship. SUBJECTS: Medical students in final (sixth) year clerkships in medicine, surgery and critical care and clinical tutors. METHOD: A self-report questionnaire on time spent in clerkship activities was administered 3 months before the final examinations. RESULTS: The amounts of time spent on current activities were fairly evenly spread over teaching, study and self-directed patient contacts, and observing patient care (ward rounds, operating theatre, accident and emergency, outpatient department and clinical meetings). Less time was available for recreation. Students and tutors concurred, independently, that good examination and internship preparation required a shifting of the balance. Examination preparation redistributed time from observing patient care and recreation to study and self-directed patient contacts. Internship preparation redistributed time from teaching and recreation to observing patient care. CONCLUSION: Students and tutors perceived that current activities were not well aligned with assessment requirements but were better aligned with the requirements of internship. If we want students to direct their attention towards curricular goals, we need to bring goals, teaching/learning activities and assessment into alignment. PMID- 12558882 TI - Directing student response to early patient contact by questionnaire. AB - CONTEXT: First year medical students experienced early patient contact by observing outpatient consultations. OBJECTIVES: To evaluate a questionnaire designed to examine emerging attitudes during the development of a doctor-patient relationship. METHODS: First year medical students participated in medical outpatient consultations as observers. These consultations represented a total of 295 registered clinical appointments. After each observation, the students completed a questionnaire surveying themes related to the doctor-patient encounter. An instructor met the students at the end of the appointments to discuss the questions raised. The Dundee Ready Education Environment Measure was used to assess the course environment. RESULTS: Students found the activity useful and enjoyable. They reported increased self-esteem and enhanced enthusiasm for the study of medicine. They completed write-ups describing and evaluating the questionnaire data. The questionnaire notes showed homogeneity among the students. The Dundee Measure indicated the students regarded the learning environment positively (62.2%); social and ambient conditions were rated highly (64.6% and 64.7%, respectively). CONCLUSION: The teaching of humanitarian attitudes by observation of the doctor-patient relationship in practice was welcomed by students. The environment in which this educational programme was carried out was considered adequate. The outpatient service schedule and the limited time available for student instruction on the part of staff doctors made this activity productive. The programme motivated students towards higher achievement and the pursuit of medical responsibility as well as more developed humanitarian behaviour patterns. PMID- 12558883 TI - Achieving acceptable reliability in oral examinations: an analysis of the Royal College of General Practitioners membership examination's oral component. AB - BACKGROUND: The membership examination of the Royal College of General Practitioners (RCGP) uses structured oral examinations to assess candidates' decision making skills and professional values. AIM: To estimate three indices of reliability for these oral examinations. METHODS: In summer 1998, a revised system was introduced for the oral examinations. Candidates took two 20-minute (five topic) oral examinations with two examiner pairs. Areas for oral topics had been identified. Examiners set their own topics in three competency areas (communication, professional values and personal development) and four contexts (patient, teamwork, personal, society). They worked in two pairs (a quartet) to preplan questions on 10 topics. The results were analysed in detail. Generalisability theory was used to estimate three indices of reliability: (A) intercase (B) pass/fail decision and (C) standard error of measurement (SEM). For each index, a benchmark requirement was preset at (A) 0.8 (B) 0.9 and (C) 0.5. RESULTS: There were 896 candidates in total. Of these, 87 candidates (9.7%) failed. Total score variance was attributed to: 41% candidates, 32% oral content, 27% examiners and general error. Reliability coefficients were: (A) intercase 0.65; (B) pass/fail 0.85. The SEM was 0.52 (i.e. precise enough to distinguish within one unit on the rating scale). Extending testing time to four 20-minute oral examinations, each with two examiners, or five orals, each with one examiner, would improve intercase and pass/fail reliabilities to 0.78 and 0.94, respectively. CONCLUSION: Structured oral examinations can achieve reliabilities appropriate to high stakes examinations if sufficient resources are available. PMID- 12558884 TI - Comparison of a rational and an empirical standard setting procedure for an OSCE. Objective structured clinical examinations. AB - PURPOSE: Earlier studies of absolute standard setting procedures for objective structured clinical examinations (OSCEs) show inconsistent results. This study compared a rational and an empirical standard setting procedure. Reliability and credibility were examined first. The impact of a reality check was then established. METHODS: The OSCE included 16 stations and was taken by trainees in their final year of postgraduate training in general practice and experienced general practitioners. A modified Angoff (independent judgements, no group discussion) with and without a reality check was used as a rational procedure. A method related to the borderline group procedure, the borderline regression (BR) method, was used as an empirical procedure. Reliability was assessed using generalisability theory. Credibility was assessed by comparing pass rates and by relating the passing scores to test difficulty. RESULTS: The passing scores were 73.4% for the Angoff procedure without reality check (Angoff I), 66.0% for the Angoff procedure with reality check (Angoff II) and 57.6% for the BR method. The reliabilities (expressed as root mean square errors) were 2.1% for Angoffs I and II, and 0.6% for the BR method. The pass rates of the trainees and GPs were 19% and 9% for Angoff I, 66% and 46% for Angoff II, and 95% and 80% for the BR method, respectively. The correlation between test difficulty and passing score was 0.69 for Angoff I, 0.88 for Angoff II and 0.86 for the BR method. CONCLUSION: The BR method provides a more credible and reliable standard for an OSCE than a modified Angoff procedure. A reality check improves the credibility of the Angoff procedure but does not improve its reliability. PMID- 12558885 TI - Clinical supervision of SpRs: where does it happen, when does it happen and is it effective? Specialist registrars. AB - OBJECTIVES: To establish what supervisory methods are used in postgraduate medical education and to determine how effective, particularly in relation to patient care, these methods are perceived to be. DESIGN: We carried out a national validated questionnaire survey of medical directors (MDs) of National Health Service (NHS) trusts (both community and hospital), educational supervisors (ESs) and specialist registrars (SpRs). SAMPLE: Seven specialties were selected to represent the potential range of supervisory practices: anaesthesia, general practice, laboratory science, medicine, paediatrics, psychiatry and surgery. These involved 15 ESs and 15 SpRs from each specialty (n = 210) and 100 MDs. MAIN OUTCOME MEASURES: Quantitative data were analysed using the following tests when appropriate: frequency counts, cross tabulations, descriptives, Kruskal-Wallis, Mann-Whitney, chi-square and Fisher exact tests. Qualitative data were also obtained. RESULTS: A total of 91% of SpRs had a formally designated ES. There was evidence of change in perceptions of the roles of supervising consultants. Supervisory practice is highly variable and there are significant differences between ESs and SpRs in perceptions of frequency and effectiveness of supervision. None of the supervisory activities, including ensuring patient safety, were rated as receiving significant or full coverage. CONCLUSIONS: Supervision is considered to be both important and effective but there is inadequate coverage and frequency of supervision activities. At the least this indicates a need for more explicit guidance for ESs and SpRs. PMID- 12558886 TI - Standards for medical educators. AB - CONTEXT: In the current UK socio-political climate of mounting regulation of professional practice, a debate on the topic of standards amongst medical teachers seems timely. The role of teacher is increasingly recognised as a core professional activity for all doctors and one that cannot be left to chance, aptitude or inclination. As a consequence, faculties have developed a plethora of teacher training programmes for medical teachers. But what is good medical teaching? Unless we know what it is, how can we develop it? One possible approach is to develop clear and comprehensive standards, defining what we mean by competent or effective clinical teaching. METHODS: In this article we have evaluated and compared two models of standards for clinical educators. The first is the outcome-based approach developed at Dundee Medical School and the second is the scholarship model devised initially by Boyer and then elaborated by Glassick and Fincher et al. The key features of both models are briefly described and their comparative strengths and problematic aspects explored. CONCLUSION: Both models offer interesting and stimulating ideas and together they provide an instructive contrast. They make a valuable contribution to the ongoing process of improving the provision of medical education. PMID- 12558887 TI - A follow-up of medical graduates of a problem-based learning curriculum. AB - INTRODUCTION: There is little information available on the effects of problem based undergraduate curricula on doctors and their performances after graduation. Therefore, we conducted a questionnaire study of all graduates of the new medical programme at the Faculty of Health Sciences, Linkoping University. METHODS: All 446 medical students who had graduated from the new programme were asked to fill in a questionnaire about selected activities during their studies and their careers after graduation. They were also asked to evaluate the quality of their undergraduate education retrospectively. Statistical analysis was performed using descriptive, multivariate and bivariate approaches. RESULTS: A total of 77% of the graduates responded. They showed a high degree of overall contentment with their undergraduate education and felt well prepared for professional life during their preregistration period and specialist education (mean = 4.0 on a 6-point Likert scale ranging from 0 to 5). They felt especially well prepared in terms of skills for communication with patients, collaboration with other health professionals and development of critical thinking/scientific attitudes. The students' age at the beginning of their studies correlated positively with their contentment as graduates, especially in terms of preparation for patient communication and collaboration with other health professionals. No differences between students originally admitted via a local admission procedure and those admitted via a national procedure were detected concerning retrospective evaluation of undergraduate medical education. CONCLUSION: Graduates of the new curriculum showed a high degree of satisfaction with their undergraduate education and its preparation of them for medical practice. Specifically, they were very content with the particular emphases of the new curriculum. PMID- 12558888 TI - Compassion fatigue: how much can I give? PMID- 12558889 TI - In search of wisdom: William Osler and the humanities. PMID- 12558891 TI - Linda Snell [interview]. PMID- 12558890 TI - Responding to the implications of the genetics revolution for the education and training of doctors: a medical humanities approach. AB - BACKGROUND: Rapid advances in the field of genetics continue to present medical educators with significant challenges. Whilst there is undoubtedly a pressing need to educate doctors about genetic disorders, research and therapies, there is a parallel need to provide a context for all of these. CONTENT: An interdisciplinary, arts and humanities based approach, responding to this need, is described. This teaching has been successfully delivered both as optional and core undergraduate teaching, and as part of continuing professional development. THE HUMAN PERSPECTIVE: STORIES NOT HISTORIES: Understanding of the patient's perspective can be significantly improved by drawing on both written and oral stories of illness. THE HISTORICAL PERSPECTIVE: LEARNING FROM THE PATIENT: Experiential learning provides insights into the social history of developments in genetics, thereby placing the current concern and debate about the new genetics in context. THE ROLE OF THE MEDIA: THE POWER TO PERSUADE: Critical reading skills can be developed and the power of the popular press to influence the reader acknowledged by analysing and employing the skills of the journalist when reporting developments in biotechnology. LEARNER ASSESSMENT AND EVALUATION: Assessment, both formative and summative, demonstrates sophisticated insights and perspectives into the lived experience of genetic illness. Learner evaluation of the teaching is high. CONCLUSION: Medical humanities offers a powerful way to convey an understanding of how genetic disorders impact on the lives of patients and families, and to set this against the background of a history rich in the uses, and abuses, of knowledge of heredity. PMID- 12558904 TI - Hepatitis B vaccine -- do we need boosters? AB - This review analyses the cumulated data from a number of long-term follow-up studies among infants, children and adults vaccinated against hepatitis B in industrialised and developing countries. Despite low or undetectable antibody responses years after vaccination, the development of HBsAg was a rarity and, if present, only transient. Some vaccinees developed anti-HBc responses but none developed an HB carrier state or clinical manifestations of disease. Studies demonstrating anamnestic responses among those with low or undetectable anti-HBs levels following challenge with HB vaccine, together with the production of anti HBs in circulating B-cells by spot ELISA, confirmed the presence of immune memory among vaccinees. Anamnestic anti-HBs responses all correlate close in kinetics and magnitude with proliferative T-cell responses. The accumulated data from studies assessed in this Review indicate that protection is dependent on immune memory, rather than declining anti-HBs responses and add additional weight to the European Consensus recommendations (12) that following a complete course of vaccination, booster doses are unnecessary in immunocompetent persons. If implemented, this recommendation will have considerable cost benefits world-wide. PMID- 12558905 TI - In vivo immunization following virus suppression: a novel approach for inducing immune control in chronic hepatitis B. AB - summary. Antiviral treatment of patients with active chronic hepatitis B may lead to significant reduction in morbidity and mortality. However, after stopping nucleoside therapy, relapse rates are high in those without acquired specific immunity. We have treated two chronic hepatitis B patients with in vivo immunization. In vivo immunization aims to optimize conditions for an effective immune response: following rapid and profound virus suppression by interferon lamivudine combination therapy, lamivudine is withdrawn intermittently for 4 weeks during continued interferon therapy. In both patients with profound virus suppression a rapid rebound in viral replication was observed after lamivudine withdrawal; despite continued interferon. These periods of renewed viral replication were followed by rises in hepatitis activity. After re-introduction of lamivudine HBV DNA became undetectable by PCR followed by normalization of serum ALT. These observations are a stimulus to further explore the concept of in vivo immunization as a novel therapeutic approach for chronic hepatitis B. PMID- 12558907 TI - The rate of fibrosis progression is an independent predictor of the response to antiviral therapy in chronic hepatitis C. AB - The response to antiviral therapy and the rate of fibrosis progression in chronic hepatitis C virus (HCV) infection are related to common factors, including age and gender. The aim of this study was to evaluate the relationship between the rate of fibrosis progression and the sustained virologic response (SVR) to interferon (IFN)-based treatment. A total of 332 patients treated for chronic HCV infection were evaluated. Using multivariate logistic regression analysis, the impact of the rate of fibrosis progression (defined as the stage of liver fibrosis according to the Metavir system/duration of infection) on the rate of SVR (negative HCV RNA at least 6 months following the end of treatment), was determined. The median age of the patients was 44 (range 25-77) years, 64% were male, and 66% were infected with genotypes 1, 4, 5 or 6. The median rate of fibrosis progression was 0.083 (range 0-2) Metavir units/year; 158 patients (48%) had F2-F4 fibrosis. After a median of 48 (range 2-126) weeks of therapy (IFN, n=190; IFN and ribavirin, n=96; pegylated IFN, n=20; pegylated IFN and ribavirin, n=26), 93 patients (28%) achieved an SVR. In univariate analysis, the rate of SVR was higher in slow [< 0.083 Metavir units/year (n=162)] than rapid progressors [> or = 0.083 Metavir units/year (n=170)] (35%vs 22%, P=0.01). After controlling for age, gender, genotype, viral load, and treatment regimen and duration, the rate of fibrosis progression remained an independent predictor of SVR (slow vs rapid progressors, OR 2.74; 95% CI 1.27-5.92; P=0.01). Hence the rate of fibrosis progression is an independent predictor of SVR to IFN-based therapy in patients with chronic hepatitis C. This additional factor should be considered in economic models evaluating this condition. PMID- 12558906 TI - Lower expression of CD81 B-cell receptor in lymphoproliferative diseases associated with hepatitis C virus infection. AB - Chronic hepatitis C virus (HCV) infection is frequently associated with type II mixed cryoglobulinaemia (MC), a benign lymphoproliferative disease (LPD). More recently, HCV has been implicated as a possible aetiologic factor of B-cell non Hodgkin lymphoma (B-NHL). CD81, a B-cell surface receptor, has been proposed as a receptor for HCV binding and entry in circulating B cells. The stimulation of CD81 complex enables B cells to respond to lower concentrations of antigen and finally induces B-cell proliferation. We studied the phenotypic expression of CD81, CD19 and CD5 on circulating B cells in HCV patients LPD-positive or LPD negative. Sixty-two patients were anti-HCV antibody positive. Among HCV positive patients, 44 were HCV RNA positive with an histologically proven chronic active hepatitis of whom 10 had a B-NHL, 14 an MC and 24 no extrahepatic manifestation. Eighteen patients were HCV RNA negative with evidence of resolved infection. A control group included 40 healthy subjects. Peripheral blood mononuclear cells (PBMC) were stained for surface expression of CD81, CD19 and CD5 using monoclonal antibodies, and were analyzed by flow cytometry. The percentage of PBMC expressing CD81, CD19 and CD5 receptors were compared between the groups by univariate analysis. Logistic regression model variables were then evaluated to correlate the presence of an LPD with HCV infection characteristics (i.e. age, gender, genotype, duration of infection, HCV RNA positivity, liver histological lesions), or phenotypic expression of CD81, CD19 and CD5 receptors on PBMC. HCV antibody-positive compared with HCV-negative subjects had a higher expression of CD19 receptor (23 +/- 13 vs 13 +/- 1%, P = 0.003). Among HCV RNA positive patients, LPD+ compared with LPD- patients had a lower expression of CD81 (58 +/- 28 vs 82 +/- 18%, P = 0.001) and CD5 receptor (66 +/- 16 vs 74 +/- 13%, P = 0.04). In multivariate analysis, the expression of CD81 receptor was a negative (OR = 0.15, 95% CI = 0.04-0.64, P = 0.01) and CD19 receptor a positive (OR = 4.81, 95% CI =1.29-17.88, P = 0.02) predictive factor for an LPD. We found two negative predictive factors for HCV RNA positivity, i.e. age (OR = 0.23, 95% CI. = 0.08-0.62, P = 0.003) and the expression of CD81 receptor (OR = 0.34, 95% CI = 0.13-0.89, P = 0.02). In patients with a chronic active HCV infection, the presence of a lymphoproliferative disease, either MC or B-NHL, is associated with lower expression of CD81 and higher expression of CD19 receptor on peripheral B cells. PMID- 12558908 TI - The long-term efficacy of plasma-derived hepatitis B vaccine in babies born to carrier mothers. AB - The long-term efficacy of a childhood hepatitis B vaccination programme was evaluated. A total of 112 newborn babies of hepatitis B carrier mothers were given hepatitis B immune globulin (HBIG) and a 10-microg three-dose regimen of plasma-derived vaccine administered at a conventional (0, 1, 6 months), delayed (2, 3, 8 months) or accelerated (0, 1, 2 months) schedule. The vaccinees were followed up to determine their anti-HBs status over a 16-year period. Upon completion of the vaccination schedules, 92.6% developed antibody against surface antigen (anti-HBs) seroconverion, the rate of which fell to 33.3% at year 16. The three schedules were equally effective in preventing chronic infection, with a protective efficacy of 88.9% from hepatitis B surface antigen (HBsAg) carriage, compared with historical control. Vaccinees on the delayed schedule had a slightly higher seroconversion rate over years, and were better able to maintain an anti-HBs level of > or = 100 iu/L. Overall, a quarter demonstrated evidence of exposure to the virus, being positive for antibody against core antigen or HBsAg, or mounting a rise in anti-HBs during the follow-up period. We conclude that a three-dose hepatitis B vaccination regimen is generally effective in protecting newborns of hepatitis B carrier mothers from infection and chronic carriage. Booster is not needed even after 16 years of monitoring. PMID- 12558909 TI - Adoptive transfer of donor-derived immunity by liver transplantation: a potential avenue to prevent hepatitis B virus reinfection. AB - Immunity to hepatitis B has been successfully transferred by bone marrow transplantation, but has also occurred after liver transplantation (LTx). This study was designed to analyse the influence of alloreactivity and immunosuppression, on the efficacy of adoptive immune transfer to hepatitis B by liver transplantation. Orthotopic LTx (n = 34) were performed in three rat strain combinations representing different genetic constellations. Donors had been vaccinated twice with recombinant hepatitis B surface antigen while recipients were unimmunized. Half of the allogeneic recipients were immunosuppressed with cyclosporin A. All animals were monitored weekly for the presence of anti hepatitis B surface antibodies (anti-HBs). Effective anti-HBs titres were detected in 85% (29/34) of liver recipients and lasted from 2 to 10 weeks. Donor titre above >15 000 mIU/mL ensured a 100% seroconversion rate in the recipients. The maximal anti-HBs titre in recipients represented 0.06% approximately 0.76% of the donor titre. Rejection reduced the adoptive immune transfer, which was protected by immunosuppression. These observations suggest that transfer of functionally active donor lymphocytes, deriving from the graft, contributed to the donor-derived immune response in the recipient. Further studies to augment the donor-derived immune response are warranted to ensure a therapeutic effect for the recipient at risk of reinfection. PMID- 12558910 TI - Early viral kinetics on treatment with pegylated interferon-alpha-2a in chronic hepatitis C virus genotype 1 infection. AB - Even with pegylated (PEG) interferons (IFN), therapy of chronic hepatitis C (genotype 1) remains unsatisfactory. The initial viral response to IFN identifies patients infected with IFN resistant viruses not responding to standard IFN/ribavirin therapy. The impact of primary IFN unresponsiveness for PEG-IFN alpha-2a/ribavirin therapy is unknown. Viral load was measured in 22 chronic hepatitis C (genotype 1) patients before and 24 h after 9 MU IFN-alpha-2a (days 0 and 1), and before and during weekly 180 microg PEG-IFN-alpha-2a (days 7, 8, 11, 14 and 21) administration. Thereafter, ribavirin (800 mg/d) was added for 6 months. Virological responders continued treatment for a further 6 months. Twenty eight patients treated with standard IFN/ribavirin therapy in a previous study using an analogous protocol served as historic controls. After 6 months 15 (68.2%) patients were (HCV-RNA) negative, eight of whom (36.4%) had a sustained response. The decrease in viral load 24 h after 9 MU IFN-alpha-2a was greater in virological responders (1.05 log [0.25-1.67]) than in nonresponders (NR) (0.34 [0.14-0.65]; P=0.003). In contrast, viral decline was not different between responders and NRs during the first 2 weeks on PEG-IFN-alpha-2a. All patients with an initial decline > 1.4 log became sustained responders. Five of 12 patients with a log change < 0.8 became end of treatment responders, two had a sustained response. Antiviral response to PEG-IFN-alpha-2a is different to that on standard IFN. In spite of a lower initial response PEG-IFN-alpha-2a/ribavirin combination therapy may overcome predicted IFN unresponsiveness. PMID- 12558911 TI - First phase hepatitis c viral kinetics in previous nonresponders patients. AB - A large proportion of patients fails to respond to treatment for hepatitis C. Initiation of interferon therapy is associated with a rapid first phase decline in viremia, reflecting inhibition of viral production or release from infected cells. We characterized first phase viral kinetics in previous nonresponder patients and compared the antiviral efficacy of interferon in nonresponders to that observed in naive patients. Twenty nonresponders with genotype 1 infection were evaluated. Ten received a single 15 mcg dose of interferon alfacon-1 and ten were given a 30 mcg dose. Viral kinetic data from previously untreated historical control patients with genotype 1 infection who received 9 mcg (n = 12) or 15 mcg (n = 13) of interferon alfacon-1 provided a basis for comparison. Antiviral efficacy was evaluated by calculating the reduction in HCV RNA levels during the first 24 h after interferon administration (log effectiveness). Hepatitis C virus levels decreased exponentially in previous nonresponder patients. Non-responders treated with 30 mcg of interferon alfacon-1 exhibited a greater log drop than non responders receiving 15 mcg (P = 0.01). The log effectiveness of 15 mcg of interferon alfacon-1 in nonresponders was similar to 9 mcg in naives and was significantly < 15 mcg (P = 0.04) in naive patients. The 30 mcg dose provided similar log effectiveness in nonresponders compared with 15 mcg in naive patients and exceeded the log effectiveness of 9 mcg in previously untreated patients (P = 0.035). Nonresponders who had greater than a 50% decrease in HCV RNA level from baseline at the end of previous treatment had a larger reduction in viral load at 24 h compared with those who had not achieved that level of response with prior therapy (P = 0.04). In conclusion, the log effectiveness of interferon was lower in nonresponders compared with treatment naive patients. The difference in antiviral effectiveness in previous nonresponders was overcome by higher interferon doses. PMID- 12558912 TI - Comparative effects of adefovir and selected nucleoside inhibitors of hepatitis B virus DNA polymerase on mitochondrial DNA in liver and skeletal muscle cells. AB - Adefovir is a potent nucleotide analog inhibitor of hepatitis B virus (HBV) DNA polymerase. Its oral prodrug adefovir dipivoxil has been approved for the treatment of chronic HBV infection. In this study, adefovir was characterized for its in vitro effects on mitochondrial DNA (mtDNA) synthesis and compared with the nucleoside analogues lamivudine (3TC), fialuridine (FIAU), and zalcitabine (ddC). No substantial changes in mtDNA content were detected in human hepatoblastoma HepG2 cells and normal human skeletal muscle cells following a 9-day treatment with 0.3-30 microm adefovir, concentrations up to 500-fold higher than the peak serum levels in patients treated with adefovir dipivoxil. Similarly, mtDNA was unchanged in both cell types following treatment with 3TC. In contrast, 30-55% and > 90% reductions in mtDNA were observed following incubation with 30 microm FIAU and ddC, respectively. The effects of FIAU on mtDNA became more pronounced following prolonged 18-day treatment of skeletal muscle cells while the effects of other drugs remained unchanged. PMID- 12558913 TI - The safety and tolerability of daily infergen plus ribavirin in the treatment of naiive chronic hepatitis C patients. AB - The treatment of chronic hepatitis C patients was enhanced when the combination of interferon alfa-2b and ribavirin was shown to be safe and more effective than interferon monotherapy. To date, no published reports have addressed the use of consensus interferon (CIFN) when combined with ribavirin. We conducted a pilot study to compare the safety and tolerability of daily CIFN plus ribavirin to CIFN monotherapy for the initial treatment of chronic hepatitis C patients. Forty subjects were randomized to two treatment groups; CIFN 9 microg daily, or CIFN 9 microg daily plus ribavirin 1000 or 1200 mg daily. All subjects received 48 weeks of therapy except for nongenotype 1 subjects in the combination treatment group who received only 24 weeks of therapy. The results show that at baseline, age, gender, risk factors, race, RNA titres, and liver histology were not different between the two groups. The proportion of subjects with genotype 1 infection was 50% (10/20) and 55% (11/20) for the monotherapy and combination therapy groups, respectively. Fifty (10/20) and sixty-five (13/20) per cent of subjects in the monotherapy and combination therapy groups exhibited a 2-log or greater decrease in viral titre at week 12 (P = NS). Using intent-to-treat analysis, 20% and 40% of enrolled subjects exhibited a sustained viral response in the monotherapy and combination therapy groups, respectively (P = NS). The proportion of subjects requiring dose reduction was 55% (11/20) and 65% (13/20), respectively. Study discontinuations for any reason were 25% (5/20) and 35% (7/20) for the monotherapy and combination groups, respectively. Discontinuations due to adverse events related to study drug were 20% (4/20) and 25% (5/20), respectively. A total of four serious adverse events occurred, two in each treatment group, only one of which was determined to be study-drug related. It is concluded that the safety and tolerability profiles of the two treatments were similar suggesting that daily dosing of CIFN may be difficult to tolerate resulting in discontinuation of therapy in a significant proportion of patients. The combination regimen resulted in a trend towards a higher viral response rate than monotherapy treatment. These data suggest that CIFN may be safely combined with ribavirin and may enhance the sustained response rate but is not well tolerated in US patients when given in a daily dosing regimen. PMID- 12558914 TI - Aetiology, clinical course and outcome of sporadic acute viral hepatitis in pregnancy. AB - Hepatitis E causes large-scale epidemics in endemic areas. The disease, during epidemics, has increased incidence and severity in pregnant women. Sporadic acute viral hepatitis (AVH) is common in endemic areas. The relationship of sporadic AVH and pregnancy has not been well studied. Over a 3-year period we prospectively studied 76 pregnant women and 337 non-pregnant women of childbearing age with sporadic acute viral hepatitis for aetiology, clinical course and outcome of disease. The aetiology in sporadic AVH was hepatitis A virus (HAV) in six (1.5%), hepatitis B virus (HBV) in 62 (15%), hepatitis C virus (HCV) in seven (1.7%), hepatitis D virus (HDV) co-infection in six (1.5%), hepatitis E virus (HEV) in 205 (49.6%), and hepatitis non-A-to-E (HNAE) in 127 (30.7%). Sixty-five (85.5%) pregnant women and 140 (41.5%) nonpregnant women had hepatitis E. The proportion of pregnant women was 31.7% in HEV group and 5.3% in non-HEV group [P < 0.001; OR=8.3 (95%C1 4.2-16.3)]. The prevalence of HEV in pregnant women in first trimester (76.9%), second trimester (88.9%), third trimester (83.8%) and puerperium (100%) did not differ significantly (P=0.09). Forty-seven (61.8%) of the 76 pregnant women developed fulminant hepatic failure (FHF), 69.2% in HEV group and 10% in non-HEV group (P < 0.001). Thirty-four (10.1%) nonpregnant women developed fulminant hepatic failure, 10% in HEV group and 9.7% in non-HEV group (P=0.86). FHF had occurred in four (40%) of 10 patients with HE in first trimester as against 41 (74.5%) of 55 patients in second trimester and beyond (P=0.015). Amongst the major complications of fulminant hepatic failure, cerebral oedema (53.2%) and disseminated intravascular coagulation (21.3%) occurred more often in pregnant women than in nonpregnant women (29.4% and 2.8%; P=0.03 and 0.016, respectively) while infections occurred more often in nonpregnant women (36.1%) than in pregnant women (10.6%; P=0.003). Fifty (61.7%) patients with FHF died [25 (53.2%) pregnant women and 25 (69.5%) nonpregnant women (P=0.06)]. Cerebral oedema and HEV aetiology were independent variables of survival in patients with FHF. Patients with cerebral oedema had worse prognosis and patients with HEV aetiology had best chances of survival. Hence HEV was the most common cause of sporadic AVH in this endemic area. High proportion of pregnant women and increased severity of disease in pregnancy were limited to patients with hepatitis E. Sporadic AVH caused by agents other than HEV did not show any special predilection to or increased severity in pregnancy. FHF in pregnant women caused by HEV was an explosive disease with short pre- encephalopathy period, rapid development of cerebral oedema and high occurrence of disseminated intravascular coagulation and may represent a severe manifestation of a Schwartzmann-like phenomenon. PMID- 12558915 TI - Targeted destruction of the polymerized human serum albumin binding site within the preS2 region of the HBV surface antigen while retaining full immunogenicity for this epitope. AB - The 55-amino acid (a.a.) preS2 region of the hepatitis B virus (HBV) envelope protein is highly immunogenic, and antibodies against this epitope confer seroprotection against HBV infections. Accordingly, various experimental and clinical studies for developing and evaluating HBV vaccines that include this particular epitope have been reported. However, a pitfall in using preS2 epitopes as part of a vaccinating antigen is that polymerized human serum albumin (pHSA), which is a normal constituent of the human serum, binds to and makes complexes with this particular region. Consequently, it is most likely that the antigen epitope is masked by serum pHSA and subsequently not detected by the immune system. To overcome these limitations, a novel single a.a substitute of the preS2 region was designed that corresponds to a tyrosine to serine exchange at position 140 of preS2. Competitive enzyme-linked immunosorbent assay showed that this substitution completely abolishes pHSA-binding activities in the mutated preS2 peptide, and CD spectra analysis revealed that this property might have been induced by slight conformational changes in its secondary structure. Nevertheless, the original B-cell epitope was still preserved in the mutated preS2 as determined by experimental immunization in mice. In this regard, the preS2(120-145/Y140S) sequence may be an HBV vaccine where epitopes, with intrinsic properties have been deleted without affecting the immunogenicity of the epitope itself. It is expected that the inclusion of this point mutated preS2 epitope will improve the efficacy of conventional preS2-containing HBV vaccines. PMID- 12558917 TI - Deaths associated with restraint use in health and social care in the UK. The results of a preliminary survey. AB - Many aspects of the management of acutely disturbed behaviour have only relatively recently come under systematic scrutiny. Perhaps regrettably one of the last amongst the range of strategies that may be employed to be subjected to rigorous examination has been physical restraint. Considerable debate has recently taken place around what represents good practice in this sensitive and controversial area but the continuing dearth of research in some aspects of this area of practice has meant that this discussion has arguably been over reliant on 'expert' opinion. Questions continue regarding some fundamental issues of restraint, including the relative risks involved in alternative approaches, and anxieties have been expressed about the potential for injuries and death to result from restraint. This article outlines the results of a survey that sought to explore the incidence of deaths associated with restraint in health and social care settings in the UK. The outcome of an initial analysis of the cases identified is then discussed, with reference to the literature on restraint related deaths, in order to identify the implications for practice. PMID- 12558919 TI - Post-psychiatry: paradigm shift or wishful thinking? A speculative review of future possibles for psychiatry. AB - Psychiatric-mental health nurses have yet to arrive at a consensus definition and agreed operational summary of their roles and responsibilities. Although the craft seems often disunited and uneasy in its traditional alliances with general nursing and psychiatry, it remains subordinate to both. This paper identifies some actual or possible developments in social values, science and technology, workplace economics and management that could impact upon the nature and practice of psychiatry, and thereby influence the possible future scope and practice of psychiatric-mental health nursing. PMID- 12558918 TI - Too tired to care? The psychological effects of working with trauma. AB - This quantitative and qualitative longitudinal study was designed to examine the effects on caregivers working therapeutically with seriously traumatized people. The participants were 13 healthcare workers seconded into a trauma and recovery team (TRT) set up to help those traumatized by the Omagh bombing on 15 August 1998. Quantitative data were collected using the Compassion Satisfaction/Fatigue Test and the Life Status Review Questionnaire. Qualitative data regarding positive and negative aspects of working with traumatized individuals, as well as caregiver's experience leaving the TRT, were gathered using open-ended questions contained in the final data set. These questionnaires were completed at four points in time: August 1998, December 1998, August 1999 and February 2001. Ethical approval for the study was gained from the Sperrin Lakeland Trust research committee. Analysis of the quantitative data indicated that levels of compassion fatigue and burnout increased, respectively, from 18.85 to 34.46 and 22.38 to 29.69 over the first year. Levels of compassion satisfaction decreased from 87.62 to 80.15, while levels of satisfaction with life (53.85 to 40.38) and life status (11.23 to 5.62) also decreased. Findings also demonstrated that compassion satisfaction is possibly a protective factor against compassion fatigue and burnout, in that caregivers with high compassion satisfaction scores were less likely to have corresponding high compassion fatigue and burnout scores. Findings from the qualitative data generated, revealed that team spirit and camaraderie, along with the satisfaction of seeing clients recover, were the most positive aspects of working in the team. Media interest, coping with and containing anger shown by bereaved relatives and dealing with the content of client's stories were the most negative. Themes to emerge from leaving the team included lack of understanding and support from non-trauma managers and an underestimation of the impact on caregivers of finishing in the team. Strategies that were found to be beneficial in helping to alleviate the negative effects of working with trauma were also elicited. This research has implications for practice, management and education. PMID- 12558920 TI - CARE: a framework for mental health practice. AB - Much has been written about care and its centrality to nursing practice. Yet nursing theories and frameworks surrounding care have either been esoteric or prescriptive and are seen as irrelevant to practice. This paper problematizes the notion of care with a view to developing a clearer, flexible framework for mental health practice. The framework is the product of the authors' shared and individual experiences and theorizing as nurses and educators and has not yet been evaluated. The CARE framework (containment, awareness, resilience and engagement) is detailed, with examples of its use in practice. Using the framework offers a way for clinicians and carers to work more strategically with clients, helping them manage and understand their condition or disability, offering support to adapt and cope, and revealing and developing new or hidden strengths to help them endure the experience and find meaning for the recovery process. Taking up the framework within the everyday practice of mental healthcare offers several benefits for clients, clinicians and mental health. PMID- 12558921 TI - The clients' experience of the community group: implications for nurse facilitators. AB - Community or large group meetings occur in many mental health units. Literature is available that describes the possible functions of such groups, but little is known about how they are actually experienced by the members, or the implications of this for nurse facilitators, especially in the current context of inpatient care. The aim of this study was to develop an understanding of the clients' experience of the daily community group in a unit that offered both an inpatient and a day patient service for adults experiencing mental health problems. The group meeting was intended to provide an opportunity for people to examine their relationships with others and discover new ways of coping with problems and stressors. The group had a changing membership, with all staff and clients present on the unit at the time expected to attend. The meeting lasted for an hour and had no predetermined agenda. Mental health nurses made up the majority of the staff membership. Semistructured interviews were conducted with four clients who had been attending the group for between 3 and 6 months. These were tape recorded, transcribed verbatim and then analysed using the phenomenological approach of Colaizzi. Three themes emerged from the data: 'Whose Responsibility?', 'Me vs. Them' and 'What Works?' These themes have implications for nurse facilitators, namely: their awareness that such issues are of importance; their ability to recognize how and when they are present; and their ability to effectively intervene. PMID- 12558922 TI - From information to knowledge: the meaning of information-giving to patients who had experienced first-episode psychosis. AB - The reform of mental health services in Finland signifies that besides the structural development of services, the development of new approaches in care is needed. Besides the other elements, information-giving has an important role when supporting outpatients' and their caregivers' everyday lives. The aim of this study was to describe how patients with first-episode psychosis experienced the information-giving and the meaning of these experiences to these patients. The study was conducted by interviewing seven patients participating in the development of the combined model of psycho-education and need-adapted models. The data were gathered by open-ended questions and analysed within the phenomenological tradition, focusing on the understanding of the patient's experiences. The results show that patients' own understanding and acceptance of the psychotic experience affected their experiences of information-giving as well as the way their experiences were met during their care. The meaning of information was experienced with feelings of worthlessness, relief, confusion or confidence and coping. The results of this study address the importance of recognizing, hearing and appreciating patients' own experiences. Information giving should be based on the patients' own experiences, appreciating them. With the information integrated into patients' own experiences they can be helped to turn to the future after the psychotic experience. PMID- 12558923 TI - Forced medication in psychiatric care: patient experiences and nurse perceptions. AB - The aims of this study were to describe: patient experiences of and nurse perceptions of patient experiences of forced medication before, during and after forced medication; patient and nurse perceptions of alternatives to forced medication; and whether patients, according to patients and nurses, retrospectively approved of forced medication. Eleven patients and nurses were interviewed about a certain situation of forced medication. Data were analysed by content analysis. The findings demonstrate that forced medication evokes a number of patient experiences according to patients and nurses. These are related to the disease, the situation of being forcibly medicated and the drug. Patients mentioned several alternatives to the forced medication, whereas nurses mentioned no alternatives. A minority of the patients, and not as many patients as the nurses' thought, retrospectively approved of the use of forced medication. It can be concluded that patients and nurses do not share the same perceptions about what patients experience when forcibly medicated. PMID- 12558924 TI - The impact on work-related stress of mental health teams following team-based learning on clinical risk management. AB - Risk management is viewed as a systematic process based on multiprofessional and multi-agency decision-making. A learning pack was developed as part of a team based learning project aiming to encourage and develop collaborative working practice. This brought different professionals and agencies working in mental health together to learn. There is little doubt that mental health practice is a source of stress for practitioners. Apart from the stress associated with managing 'risky' situations, risk management is also a relatively new concept. This can increase stress around ability to cope, both on an individual practitioner level and in teams. This article reports the impact that the learning pack had on team members' stress, specifically work-related stress. A range of scales were used to measure change in stress and results demonstrated reduced work-related pressure in a number of areas following the learning. The implications for team learning in relation to clinical risk management are discussed in light of the findings. PMID- 12558925 TI - Early intervention in psychosis: who wins, who loses, who pays the price? AB - The following article discusses Early Intervention in Psychosis services. It is contextualized with the rapidly changing psychiatric system. There is exploration of the reasons for a focus upon Early Intervention in Psychosis at this current time. The article then proceeds to discuss what may be needed for services to meet government targets. It concludes with some tentative answers and further questions to issues raised within the discussion. PMID- 12558926 TI - Meanings of madness: a literature review. AB - This literature review focuses on how people make sense of mental illness. The study explores the process of meaning making by people experiencing mental 'dis order' as it is influenced by the context of their environments, available cultural explanations and the nature of their mental states at that time. It is proposed that people experiencing psychological trauma attempt to regain a sense of order by imposing a narrative structure on these disconcerting experiences. The sharing of these personal stories with others results in meanings being shaped and refined through dialogue and across time. Socially agreed meanings, however, can have a political dimension and it is argued that personal meanings embedded in narratives of mental pain are often suppressed and categorized to fit dominant cultural or biomedical explanations. Implications for mental health nursing are discussed. PMID- 12558927 TI - Staff morale in the merger of mental health and social care organizations in England. AB - Following the closure of the last Victorian asylum in Somerset, the health authority and county council undertook a review of mental health services. A major outcome of this review was the creation of an integrated mental health and social care provider. The current paper explores the impact of this integration on the morale of staff members involved, using a conceptual model derived from the literature on organizational behaviour. During the year immediately following integration, the average ratings on all measures of role clarity and job satisfaction reduced. For staff members involved in the integration, by far the largest group of whom were mental health nurses, job satisfaction was related to team role clarity, team identification, emotional exhaustion and gender. These effects of the integration on staff morale are discussed in light of the wider research into the determinants of job satisfaction and the conditions for success in merging organizations. The study has significant implications for managerial and professional leadership during organizational change. PMID- 12558928 TI - The perceived expressed emotion in staff scale. AB - Recent research has highlighted the role of expressed emotion by ward staff in determining the well-being of psychiatric inpatients. Existing methods of assessing staff expressed emotion involve standardized interviews and are expensive and time-consuming. We report the development of a questionnaire measure of expressed emotion in staff as perceived by patients. In study 1, factor analysis of items administered to patients in a variety of settings led to the development of a questionnaire with three subscales: supportiveness, criticism and intrusiveness. In study 2, the test-retest reliability of the questionnaire was found to be adequate, and some evidence of concurrent validity for the scale was obtained against expressed emotion rated from staff speech samples. In study 3, the scale was shown to have good concurrent validity against the ward atmosphere scale, and scores were found to be independent of insight or experience of admission to hospital. The perceived expressed emotion in staff scale is a convenient measure, which may have utility for both research and clinical purposes. PMID- 12558929 TI - Conference report. PMID- 12558931 TI - Participation of endogenous IGF-I and TGF-beta 1 with enamel matrix derivative stimulated cell growth in human periodontal ligament cells. AB - Previous studies have provided the biological basis for the therapeutic use of enamel matrix derivative (EMD) at sites of periodontal regeneration. A purpose of this study is to determine effects of EMD on cell growth, osteoblastic differentiation and insulin-like growth factor-I (IGF-I) and transforming growth factor-beta 1 (TGF-beta 1) production in human periodontal ligament cells (HPLC). We also examined participation of endogenous IGF-I and TGF-beta 1 with EMD stimulated cell growth in these cells. HPLCs used in this study were treated with EMD alone or in combination with antihuman IGF-I antibody (anti-hIGF-I) or anti hTGF-beta 1, recombinant human bone morphogenetic protein-2 (rhBMP-2), 1,25 dihydroxyvitamin D3[1,25(OH)2D3], rhTGF-beta 1 or rhIGF-I. After each treatment, cell growth, the production of IGF-I and TGF-beta 1 and the expression of osteoblastic phenotypes were evaluated. EMD stimulated cell growth in dose dependent and time-dependent manners. EMD was also stimulated to express IGF-I and TGF-beta 1 at protein and mRNA levels. The EMD-stimulated cell growth was partially suppressed by cotreatment with anti-hIGF-I or anti-hTGF-beta 1, and cell growth was also stimulated by treatment with rhIGF-I or rhTGF-beta 1. rhBMP 2 stimulated alkaline phosphatase (ALPase) activity and ALPase mRNA expression, and 1,25(OH)2D3 stimulated ALPase and osteocalcin mRNA expression. However, EMD showed no effect on the osteoblastic phenotypes expression. These results demonstrated that EMD has no appreciable effect on osteoblastic differentiation, however it stimulates cell growth and IGF-I and TGF-beta 1 production in HPLC, and that these endogenous growth factors partially relate to the EMD-stimulated cell growth in HPLC. PMID- 12558932 TI - Different distribution of immunocompetent cells in the dentogingival junction during root formation in rat molars. AB - The distribution of immunocompetent cells in the dentogingival junction of rat molars during root formation was investigated by immunocytochemistry using antibodies to class II major histocompatibility complex (MHC) molecules (OX6 antibody) and monocyte/macrophage lineage cells (ED1-antibody) as well as by histochemical reaction for periodic acid-Schiff (PAS). Two portions (the junctional epithelium in the mesial gingiva of the first molar, and the interdental gingiva between the first and second molars) were selected for observations. At the eruption stage of the first molar (16-18 days after birth), OX6-positive cells, dendritic or oval in shape, were abundantly distributed in the connective tissue between the oral epithelium and tooth germ. Positive cells with slender cell processes were also found beneath the ameloblast layer. At the commencement stage of the first molar occlusion (24-28 days after birth), numerous OX6-positive cells displaying a dendritic fashion existed preferentially in the mesial gingiva, but were fewer in the interdental gingiva. In contrast, the interdental gingiva showed a denser distribution of ED1-positive cells and PAS-reactive polymorphonuclear leukocytes (PMLs) than the mesial gingiva. At the completion stage of root formation (100-120 days after birth), the OX6 immunopositive cells invaded the deeper position of the mesial gingiva with the downgrowth of the epithelium; they had a considerably higher cell density compared with those in the interdental gingiva where PAS-reactive PMLs persisted. These findings indicated that the immunocompetent cells showed a region-specific distribution and cell density by their roles in immune response. PMID- 12558933 TI - Influence of interleukin-1 gene polymorphism on periodontal regeneration in intrabony defects. AB - The aim of this controlled retrospective study was to evaluate the influence of an IL-1 gene polymorphism on the clinical and radiographic healing outcomes of GTR therapy. The study included 47 adult periodontitis patients with 94 deep intrabony defects treated by GTR using different membrane materials. The following clinical parameters were recorded at baseline and 12 months after surgery: papillary bleeding index (PBI), gingival recession (REC), probing pocket depth (PPD), clinical attachment level (CAL), and the vertical relative attachment gain (V-rAG). Bone changes in the defect regions due to GTR therapy were quantitatively evaluated using digital subtraction radiography (DSR). Polymorphisms of the IL-1A gene at position - 889 and of the IL-1B gene at position + 3953 were analyzed by PCR. Statistical analysis was performed using the Mann-Whitney-U and the Wilcoxon-Signed-Rank tests (alpha = 0.05). The study comprised 19 IL-1 genotype positive (IL-1 +) patients and 28 IL-1 genotype negative (IL-1 -) patients. Twelve months after GTR therapy, both patient groups revealed statistically significant PPD reductions and CAL gain [median (25/75% percentiles)]: Delta PPD [IL-1 + : 4.0 (2.5/5.0) mm; IL-1-: 3.8 (3.0/4.9) mm], Delta CAL [IL-1 + : 3.5 (3.0/4.8) mm; IL-1 -: 3.0 (1, 2/4, 5) mm]. V-rAG amounted to 60.0 (47.7/78.6)% in IL-1 + patients and 53.1 (43.4/81.9)% in IL-1 - patients. Both patient groups showed significant bone density gain in 40% (IL-1 +) and 43.6% (IL-1 -) of the initial defect area due to GTR. Neither the clinical nor the radiographic healing parameters revealed any statistically significant differences in the GTR healing outcome between IL-1 + and IL-1 - patients. In conclusion, these 12-month findings indicate that the IL-1 gene polymorphism has no influence on the clinical and radiographic regeneration results following GTR therapy. PMID- 12558934 TI - Determination of eight selected periodontal pathogens in the subgingival plaque of maxillary first molars in Japanese school children aged 8-11 years. AB - To prevent the onset or progression of periodontitis, we must understand when periodontal pathogens are first harbored and how they develop the biofilm that causes periodontal disease. The purpose of this study was to determine the relationship between clinical status and selected periodontal pathogens in subgingival plaque in school children. This study was conducted with 95 school children, 8-11 years old. The presence and distribution of eight selected periodontal pathogens sampled from the maxillary right first molar were determined by an indirect immunofluorescent technique and compared with clinical parameters. Of the 95 sites sampled, only one site had all eight pathogens and five sites did not have any of the eight pathogens. The mean number of positive pathogens per site was 3.5 +/- 1.8 and mean percentage of positive pathogens was 3.82 +/- 4.22%. The number and total percentage of positive pathogens were strongly correlated with the Plaque Index (PI). In addition, the number of positive pathogens was correlated to the presence of subgingival calculus. The most frequently found pathogens were Campylobacter rectus (84.2%) and Eikenella corrodens (83.2%), and the least, Actinobacillus actinomycetemcomitans serotype c (7.4%). Of the eight pathogens, the frequency and distribution of Porphyromonas gingivalis were significantly correlated with PI and the presence of calculus. In addition, seven sites with both P. gingivalis and Bacteroides forsythus showed a correlation with gingival inflammation. In conclusion, the presence of P. gingivalis or P. gingivalis and B. forsythus may be a risk marker to be sought in screening for the onset of periodontal disease. PMID- 12558935 TI - Effects of CD14 receptors on tissue reactions induced by local injection of two gram-negative bacterial lipopolysaccharides. AB - Lipopolysaccharide (LPS) was recognized by CD14, which may be an important mediator in the deleterious effects of LPS on the periodontal destruction. To investigate the roles of CD14 molecules on LPS-induced soft tissue inflammation and bone destruction, the tissues of CD14-deficient mice were examined histopathologically following a local injection of either Salmonella minnesota or Porphyromonas gingivalis LPS. In the first group, 12 mice received a local injection of 500 microg of purified P. gingivalis LPS and six mice were injected with saline to the calvaria as controls. In the second group 13 mice were injected subcutaneously on the laterally abdominal skin with 50 microg of S. minnesota LPS and three mice were injected with PBS. Mice were sacrificed at day 5. After histological preparation, the tissue sections of calvaria and soft tissue specimen were stained with tartrate-resistant acid phosphatase (TRAP) marker for osteoclast and macrophage. The soft tissue sections were also stained with hematoxylin & eosin (H&E). Resorption surface and osteoclast index were measured to quantify bone resorption. Necrotic area and inflammatory cell numbers were estimated to assess the situation of local inflammation. Our results indicated that LPS-induced bone resorption is inhibited in CD14-deficient mice. An increase in the number of total inflammatory cells was noticed in both CD14 deficient mice and wild-type mice; however, the cell numbers were less in CD14 deficient mice than those in wild-type mice (two- to three-fold decrease). Therefore, we conclude that the LPS-stimulated bone resorption is mainly via CD14 receptor but the LPS-induced soft tissue inflammation appears to be partially dependent on the receptor. PMID- 12558936 TI - Induction of c-fos expression by nicotine in human periodontal ligament fibroblasts is related to cellular thiol levels. AB - Cigarette smoking is associated with increased incidence of periodontal disease and poor response to periodontal therapy. Several studies have shown the detrimental effects of nicotine on periodontal tissue. To investigate the molecular toxicological implications of cigarette smoking on periodontal tissue, expression of c-fos early stress response gene was examined in human periodontal ligament fibroblasts (PDLFs) after exposure to nicotine. The exposure of quiescent human PDLFs to nicotine resulted in the induction of c-fos mRNA expression. The levels of the c-fos mRNAs increased about 2.5 and 4.8-fold after exposure to 2.5 mm and 10 mm nicotine for 2 h, respectively. Moreover, the peak of c-fos mRNA levels induced by nicotine was 5 mm at 2-h incubation period. Kinetic investigations of c-fos mRNA expression in nicotine-treated cells revealed a rapid accumulation of the transcript, a significant signal first detectable after 30 min of exposure. This increase was transient and the level of c-fos mRNAs returned rapidly to that of control cells by 8 h. To determine whether thiol levels were important in induction of c-fos by nicotine, we pretreated cells with the glutathione (GSH) precursor, 2-oxothiazolidine-4 carboxylic acid (OTZ), to boost thiol levels, or buthionine sulfoximine (BSO) to deplete GSH. Our results demonstrate that OTZ pretreatment decreased in c-fos mRNA level and BSO pretreatment enhanced in c-fos mRNA level after exposure to nicotine. In addition, nicotine significantly depleted intracellular GSH in a dose-dependent manner (P < 0.05). At a concentration of 5 mm and 20 mm, nicotine depleted about 22.2% and 56% of GSH, respectively. Taken together, c-fos gene expression might be one signal transduction pathway linked to the induction of early response genes by cigarette smoking. These results suggest that the nicotine-dependent stress-specific expression of the c-fos gene correlates with cellular thiol levels in human PDLFs. PMID- 12558938 TI - The influence of membrane exposure on the outcomes of guided tissue regeneration: clinical and microbiological aspects. AB - The aim of the present study was to evaluate the influence of membrane exposure on guided tissue regeneration (GTR). Thirty patients with 2-wall or 3-wall intraosseous defects were treated with GTR. Periodontal index, gingival index, bleeding on probing, probing depth, clinical attachment level and recession were assessed at the baseline examination, as well as immediately prior to and six months after GTR. The numbers of Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans on the retrieved expanded polytetrafluoroethylene (e-PTFE) membranes were assessed using polymerase chain reaction. The results showed that 26.3% of the membrane exposure sites had zero attachment gain. The membrane exposure sites tended to achieve a smaller clinical attachment gain and had significantly greater marginal tissue recession ( p = 0.006). In addition, the exposed membranes harbored significantly more A. actinomycetemcomitans than the non-exposed membranes (p = 0.029). In conclusion, membrane exposure resulted in a poor GTR outcome. The amount of A. actinomycetemcomitans on the exposed membrane may be a major contributing factor to the outcome of GTR. PMID- 12558937 TI - Immunohistochemical characteristics of epithelial cell rests of Malassez during cementum repair. AB - To clarify the roles of epithelial cell rests of Malassez (ECRM) during periodontal repair, experimental root resorption was induced in rats and then the ECRM that existed in periodontal ligament during cementum repair was investigated using morphological and immunohistochemical approaches. At day 7, after mechanical injury, root resorption was observed and ECRM were present adjacent to the site of resorption lacunae. They were observed in periodontal ligament adjacent to site of the resorption lacunae. These ECRM were immunoreactive for bone morphogenetic protein-2. During the stage of early cementum repair, the ECRM were immunoreactive for osteopontin and ameloblastin. They strongly reacted to proliferating cell nuclear antigen. In uninjured control sections, ECRM located in the periodontal ligament adjacent to cementum were not immunoreactive for any antibodies. These findings suggested that ECRM may be related to cementum repair by activating their potential to secrete matrix proteins which have been expressed in tooth development. PMID- 12558939 TI - Detection of Campylobacter rectus in periodontitis sites by monoclonal antibodies. AB - Campylobacter rectus, a gram-negative, microaerophilic, and motile bacterium, has been proposed to play a pathogenic role in human periodontitis. Surface components, such as the flagellum, surface layer (S-layer), and cytotoxin, have been reported as possible virulence factors of the microorganism. In the present study, monoclonal antibodies against surface components of this bacterium were produced to detect and investigate the pathogenic potential of C. rectus in periodontitis. Two monoclonal antibodies, designated CRT-1 and CRT-2, recognized a peculiar 150 kDa S-layer protein by immunoblot analysis. The CRT-2 antibody reacted to all C. rectus strains tested, except for the S-layer negative strain of the species [C. rectus ATCC 33238 S-layer (-) strain]. The CRT-3 antibody reacted to a 60-kDa protein in C. rectus and also cross-reacted with Campylobacter showae ATCC 51164 and CCUG 11641 strains. Using the dot-blot method, we were able to detect C. rectus using the CRT-2 antibody when as few as 103 organisms were present in a subgingival dental plaque sample. Detection of C. rectus in plaque samples correlated significantly with clinical findings such as probing depth (P < 0.001), bleeding on probing (P < 0.001), and gingival index (P < 0.001). These findings indicate that infection by C. rectus may be an important indicator of periodontal disease status. PMID- 12558940 TI - Interleukin-8 and anti-interleukin-8 autoantibodies in gingival crevicular fluid from patients with periodontitis. AB - Interleukin-8 (IL-8) is believed to play an important role in the pathogenesis of various forms of periodontitis. In addition, the anti-IL-8 autoantibody has been recently recognized as a potent modulator of IL-8 function. In the current study, the concentrations of IL-8 and its autoantibody in gingival crevicular fluid from patients with chronic generalized periodontitis were compared to those in gingival crevicular fluid from patients with refractory chronic periodontitis. Gingival crevicular fluids were collected from patients treated in a private periodontal clinic. Nine patients who were identified as having chronic generalized periodontitis and four with refractory chronic periodontitis were selected for the study. Patients included in the latter group had undergone supportive periodontal therapy for more than 10 years, and during that time had experienced many episodes of periodontal destruction. The gingival crevicular fluid concentrations of total protein, IL-8, free anti-IL-8 autoantibody and IL-8 bound to the autoantibody (anti-IL-8:IL-8 complexes) were examined. There were no differences in concentration of total protein, but significantly higher levels of IL-8 were detected in patients with chronic generalized periodontitis in comparison to patients with refractory chronic periodontitis (P < 0.05). In addition, anti-IL-8:IL-8 complexes were present in 90% of patients with chronic generalized periodontitis, but in only 50% of patients with refractory chronic periodontitis. The results suggest that elevated concentrations of free and complexed IL-8 can differentiate patients with chronic generalized periodontitis from patients with refractory chronic periodontitis. PMID- 12558941 TI - Muscarinic acetylcholine receptors regulating cell cycle progression are expressed in human gingival keratinocytes. AB - We have previously reported the presence in human gingival keratinocytes (GKC) of choline acetyltransferase, the acetylcholine (ACh) synthesizing enzyme, acetylcholinesterase, the ACh degrading enzyme, and alpha 3, alpha 5, alpha 7, beta 2 as well as alpha 9 nicotinic ACh receptor subunits. To expand the knowledge about the role of ACh in oral biology, we investigated the presence of the muscarinic ACh receptor (mAChR) subtypes in GKC. RT-PCR demonstrated the presence of m2, m3, m4, and m5 mRNA transcripts. Synthesis of the respective proteins was verified by immunoblotting with the subtype-specific antibodies that revealed receptor bands at the expected molecular weights. The antibodies mapped mAChR subtypes in the epithelium of human attached gingiva and also visualized them on the cell membrane of cultured GKC. The whole cell radioligand binding assay revealed that GKC have specific binding sites for the muscarinic ligand [3H]quinuclidinyl benzilate, Bmax = 222.9 fmol/106 cells with a Kd of 62.95 pM. The downstream coupling of the mAChRs to regulation of cell cycle progression in GKC was studied using quantitative RT-PCR and immunoblotting assays. Incubation of GKC for 24 h with 10 micro m muscarine increased relative amounts of Ki-67, PCNA and p53 mRNAs and PCNA, cyclin D1, p21 and p53 proteins. These effects were abolished in the presence of 50 micro m atropine. The finding in GKC of mAChRs coupled to regulation of the cell cycle progression demonstrate further the structure/function of the non-neuronal cholinergic system operating in human oral epithelium. The results obtained in this study help clarify the role for keratinocyte ACh axis in the physiologic control of oral gingival homeostasis. PMID- 12558942 TI - Enhanced expression of vascular endothelial growth factor by periodontal pathogens in gingival fibroblasts. AB - Vascular endothelial growth factor (VEGF) has recently attracted attention as a potent inducer of vascular permeability and angiogenesis. Aberrant angiogenesis is often associated with lesion formation in chronic periodontitis. The aim of the present study was to investigate the properties of VEGF expression in human gingival fibroblasts (HGF) culture. HGF were stimulated with lipopolysaccharide (LPS), vesicle (Ve) and outer membrane protein (OMP) from Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. HGF constitutively produced VEGF and levels were significantly enhanced (P < 0.01) by stimulation with Ve and OMP from A. actinomycetemcomitans and P. gingivalis at concentrations of 10 microg/ml or higher. On the other hand, VEGF levels were not increased by LPS stimulation. VEGF mRNA expression was also observed in Ve- and OMP-stimulated HGF. A vascular permeability enhancement (VPE) assay was performed using guinea pigs to ascertain whether supernatant from cultures of Ve- and OMP-stimulated HGF enhance vascular permeability in vivo. Supernatant from cultures of Ve- and OMP stimulated HGF strongly induced VPE. This was markedly suppressed upon simultaneous injection of anti-VEGF polyclonal antibodies with the supernatant. Heating and protease treatment of the stimulants reduced the VEGF enhancing levels in Ve and OMP in vitro. These results suggest that Ve and OMP may be crucial heat-labile and protease-sensitive components of periodontal pathogens that enhance VEGF expression. In addition, VEGF might be associated with the etiology of periodontitis in its early stages according to neovascularization stimulated by periodontal pathogens causing swelling and edema. PMID- 12558943 TI - Recombinant human basic fibroblast growth factor (bFGF) stimulates periodontal regeneration in class II furcation defects created in beagle dogs. AB - Several growth factors (or cytokines) have been recently investigated for their use as potential therapeutics for periodontal tissue regeneration. The objective of this study was to evaluate periodontal tissue regeneration, including new bone and cementum formation, following topical application of recombinant basic fibroblast growth factor (bFGF, FGF-2) to furcation class II defects. Twelve furcation class II bone defects were surgically created in six beagle dogs, then recombinant bFGF (30 micro g/site) + gelatinous carrier was topically applied to the bony defects. Six weeks after application, periodontal regeneration was analyzed. In all sites where bFGF was applied, periodontal ligament formation with new cementum deposits and new bone formation was observed histomorphometrically, in amounts greater than in the control sites. Basic FGF applied sites exhibited significant regeneration as represented by the new bone formation rate (NBR) (83.6 +/- 14.3%), new trabecular bone formation rate (NTBR) (44.1 +/- 9.5%), and new cementum formation rate (NCR) (97.0 +/- 7.5%). In contrast, in the carrier-only sites, the NBR, NTBR, and NCR were 35.4 +/- 8.9%, 16.6 +/- 6.2%, and 37.2 +/- 15.1%, respectively. Moreover, no instances of epithelial down growth, ankylosis, or root resorption were observed in the bFGF applied sites examined. The present results indicate that topical application of bFGF can enhance considerable periodontal regeneration in artificially created furcation class II bone defects of beagle dogs. PMID- 12558944 TI - Relationship between fibrinolytic activity and gingival inflammatory reaction in young individuals. AB - The fibrinolytic system (the plasminogen activating system) is involved in several physiological and pathological processes. Through the transformation of plasminogen to the aggressive broad spectrum protease plasmin, potent enzymatic activity is released. Plasmin acts directly on connective tissue components, and indirectly by activating proforms of the metalloproteinases. The destructive potential of the fibrinolytic system may thus be of importance for the initiation and progression of periodontal diseases. Earlier studies have shown high concentrations of the plasminogen activator t-PA and its inhibitor PAI-2 in gingival crevicular fluid (GCF) as well as enhanced concentrations in areas of gingival inflammation. The aim of this study was to investigate a possible relationship between the gingival inflammatory reactivity and the fibrinolytic activity in gingival crevicular fluid. Thirty-one young individuals took part in the study. Gingival Index scores and Plaque Index scores were assessed and used to formulate a score expressing an individuals' inflammatory response to microbial plaque levels (Relative G/P score). The fibrinolytic activity of GCF was assessed with a fibrin gel lysis assay, and the levels of t-PA and PAI-2 were assayed with ELISAs. All samples showed fibrinolytic activity. A positive correlation between the fibrinolytic activity and Relative G/P score was found. Thus, in individuals with an enhanced reactivity to dental plaque, a higher plasminogen activating activity in GCF was seen. This indicates a higher potential for tissue proteolysis in these individuals, possibly facilitating spread and deeper involvement of the lesions. PMID- 12558945 TI - Probing the conformational and dynamical effects of O-glycosylation within the immunodominant region of a MUC1 peptide tumor antigen. AB - MUC1 mucin is a large transmembrane glycoprotein, the extracellular domain of which is formed by a repeating 20 amino acid sequence, GVTSAPDTRPAPGSTAPPAH. In normal breast epithelial cells, the extracellular domain is densely covered with highly branched complex carbohydrate structures. However, in neoplastic breast tissue, the extracellular domain is under-glycosylated, resulting in the exposure of a highly immunogenic core peptide epitope (PDTRP in bold above), as well as in the exposure of normally cryptic core Tn (GalNAc), STn (sialyl alpha2-6 GalNAc) and TF (Gal beta1-3 GalNAc) carbohydrates. Here, we report the results of 1H NMR structural studies, natural abundance 13C NMR relaxation measurements and distance-restrained MD simulations designed to probe the structural and dynamical effects of Tn-glycosylation within the PDTRP core peptide epitope. Two synthetic peptides were studied: a nine-residue MUC1 peptide of the sequence, Thr1-Ser2 Ala3-Pro4-Asp5-Thr6-Arg7-Pro8-Ala9, and a Tn-glycosylated version of this peptide, Thr1-Ser2-Ala3-Pro4-Asp5-Thr6(alphaGalNAc)-Arg7-Pro8-Ala9. The results of these studies show that a type I beta-turn conformation is adopted by residues PDTR within the PDTRP region of the unglycosylated MUC1 sequence. The existence of a similar beta-turn within the PDTRP core peptide epitope of the under glycosylated cancer-associated MUC1 mucin protein might explain the immunodominance of this region in vivo, as the presence of defined secondary structure within peptide epitope regions has been correlated with increased immunogenicity in other systems. Our results have also shown that Tn glycosylation at the central threonine within the PDTRP core epitope region shifts the conformational equilibrium away from the type I beta-turn conformation and toward a more rigid and extended state. The significance of these results are discussed in relation to the possible roles that peptide epitope secondary structure and glycosylation state may play in MUC1 tumor immunogenicity. PMID- 12558946 TI - The CEC behaviour of several synthetic peptides related to the activin betaA betaD subunits. AB - The resolution of several structurally related synthetic peptides, derived from the loop 3 region of the activin betaA-betaD subunits, has been studied using capillary electrochromatography (CEC) with Hypersil n-octadecylsilica as the sorbent. The results confirm that the CEC migration of these peptides can be varied in a charge-state-specific manner as the properties of the background electrolyte, such as pH, salt concentration and content of organic modifier, or temperature are systematically changed. Acidic peptides followed similar trends in retention behaviour, which was distinctly different to that shown by more basic peptides. The CEC separation of these peptides with the Hypersil n octadecyl-silica involved distinguishable contributions from both electrophoretic mobility and chromatographic retention. Temperature effects were reflected as variations in both the electro-osmotic flow and the electrophoretic mobility of the peptides. When the separation forces acting on the peptides were synergistic with the electro-osmotic flow, as, for example, with the positively charged peptides at a particular pH and buffer electrolyte composition, their retention coefficient, kappacec, decreased with increasing capillary temperature, whereas when the separation forces worked in opposite directions, as for example with negatively charged peptides, their kappacec values increased slightly with increasing temperature. Moreover, when the content of organic modifier, acetonitrile, was sufficiently high, e.g. > 40% (v/v) and nonpolar interactions with the Hypersil n-octadecyl-silica sorbent were suppressed, mixtures of both the basic and acidic synthetic peptides could be baseline resolved under isocratic conditions by exploiting the mutual processes of electrophoretic mobility and electrostatic interaction. A linear relationship between the ln kappacec values and the volume fractions, psi, of the organic modifier over a limited range of psi-values, was established for the negatively charged peptides under these isocratic conditions. These findings thus provide useful guidelines in a more general context for the resolution and analysis of structurally related synthetic peptides using CEC methods. PMID- 12558947 TI - Circular dichroism and Fourier transform infrared spectroscopic studies on self assembly of tetrapeptide derivative in solution and solvated film. AB - Aggregation of the hydrophobic peptide derivative Boc-Ala-Ile-Ile-Gly-OMe (1) was examined in methanol solution and in solvated film states. Formation of the peptide by self-assembly was evidenced using fluorescence [Mg salt of 8-anilino naphthalenesulfonic acid (ANS) as an external probe] and circular dichroism (CD) spectroscopic techniques. In solution, peptide 1 formed as a stable aggregate at a concentration around 3 x 10(-4)m. The peptide gelled into a thin film for which we carried out CD and Fourier transform infrared (FTIR) measurements. Our spectroscopic study on peptide films at differing methanol concentrations indicates that the helical content of the peptide decreases with decreasing methanol concentration in solvated films. However, by reducing the methanol concentration we were able to observe a conformational transition from a predominantly helical turn to a beta-sheet structure via a random coil conformation. Our study focused on the aggregation of the alpha-helical turn forming peptide derivative, which shows conformational transition on changing solvent concentration in the film form. PMID- 12558948 TI - Relative hydrophobicity of di- to hexapeptides as measured by aqueous two-phase partitioning. AB - Partitioning of 153 di- to hexapeptides in an aqueous dextran-PEG two-phase system containing 0.15 m NaCl in 0.01 m sodium phosphate buffer, pH 7.4 was examined. The relative hydrophobicity of the peptides was estimated and expressed in equivalent numbers of methylene units. Analysis of the data shows that the additivity principle applies for the relative hydrophobicity of up to hexapeptides. The relative hydrophobicities of Trp, Glu, and Asp residues in heterooligopeptides are noticeably different from those in corresponding homooligopeptides. The relative hydrophobicity of peptides can be calculated and used as a structural descriptor in quantitative structure-activity relationship analysis. The peptide bitterness threshold is shown to be quantitatively related to the peptide structure described as a combination of the relative hydrophobicity and lipophilicity (logD) of peptides. PMID- 12558949 TI - A convenient incorporation of conformationally constrained 5,5-dimethylproline into the ribonuclease A 89-124 sequence by condensation of synthetic peptide fragments. AB - The presence of l-5,5-dimethylproline (dmP) within an amino acid sequence results in the formation of an X-dmP peptide bond predominantly locked in a cis conformation. However, the common use of this unnatural amino acid has been hampered by the difficulty of the economical incorporation of the dmP residue into longer peptide segments due to the steric hindrance imposed by the dimethyl moieties. Here, we describe synthesis of the C-terminal 36-residue peptide, corresponding to the 89-124 sequence of bovine pancreatic ribonuclease A (RNase A), in which dmP is incorporated as a substitute for Pro93. The peptide was assembled by condensation of protected 5- and 31-residue peptide fragments, which were synthesized by solid-phase peptide methodology using fluorenylmethyloxycarbonyl chemistry. We focused on optimizing the synthesis of the Fmoc-Ser(tBu)-Ser(tBu)-Lys(Boc)-Tyr(tBu)-dmP-OH pentapeptide (residues 89-93) with efficient acylation of the sterically hindered dmP residue. In a comparative study, the reagent O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate was found to be superior to bromo-tris-pyrrolidino phosphonium hexafluorophosphate and tetramethylfluoroformamidinium hexafluorophosphate for the synthesis of the -Tyr(tBu)-dmP- peptide bond in solution as well as on a resin. PMID- 12558952 TI - Invasive front grading: reliability and usefulness in the management of oral squamous cell carcinoma. AB - BACKGROUND: The value of histological grading was examined with emphasis on reliability of assessment in 102 cases of intraoral squamous cell carcinoma from Northern Ireland with known outcome. METHODS: Two pathologists independently graded the invasive tumour front blinded to the stage and outcome. RESULTS: Intraobserver agreement was acceptable but interobserver agreement was not satisfactory. The degree of keratinisation was assessed most consistently while nuclear polymorphism was the least reliable feature. Multivariate survival analysis showed that the total grading score was associated with overall survival while the pattern of tumour invasion was the most valuable feature in estimating regional lymph node involvement. The number of positive lymph nodes was strongly associated with regional relapse, while the treatment modality and status of the surgical margins correlated with local relapse. CONCLUSIONS: Grading of selected features in OSCC is reliable and can facilitate treatment planning. PMID- 12558950 TI - Synthesis of cyclic peptides from unprotected precursors using removable N alpha (1-(4-methoxyphenyl)-2-mercaptoethyl) auxiliary. AB - A new method to cyclize unprotected peptides is presented. The method involves the use of a 1-phenyl-2-mercaptoethyl derivative on the N-terminal glycine. This template acts as an auxiliary thiol-containing group in order to drive cyclization with a counterpart thioester moiety on the same molecule. Subsequent facile removal of the derivative generates products with only native peptide structure. The successful, high-yield cyclization of the peptide GSPYSSDTTPA is described. PMID- 12558953 TI - Jaw bone remodeling at the invasion front of gingival squamous cell carcinomas. AB - BACKGROUND: It is still unknown how jaw bone remodeling occurs at actual invasion sites of oral squamous cell carcinomas. Since there is no other human carcinomas which make a direct invasion of the bone, gingival carcinomas are valuable examples. METHODS: Twelve surgical specimens of gingival squamous cell carcinoma were examined histopathologically and immunohistochemically for remodeling of bone and its surrounding tissue. RESULTS: Three types of bone interfaces with carcinomatous invasion were distinguished. These included areas with bone resorption, smooth bone surface and new bone formation. In the bone-resorption area, numerous osteoclasts were located along the bone surface, which was surrounded by myxoid stroma. The myxoid stroma was characterized by immunopositivity for heparan sulfate proteoglycan (HSPG), abundant vascularity and macrophagic infiltration. In the bone-formation area, rows of osteoblasts were aligned on the bone surface. The stroma around osteoblasts was also HSPG immunopositive, poor in vascularity but rich in activated fibroblasts. In the smooth-bone area, the stroma showed an organizing phase of granulation tissue with slender fibroblasts and mature collagen fibers but with less vascularity and inflammatory infiltrates. CONCLUSION: The results indicate that the stromal architecture, especially in terms of its inflammatory cellular, vascular and matrix compositions, is strictly regulated in the timing and site of jaw bone remodeling which is causes by carcinomatous invasion. PMID- 12558954 TI - Autofluorescence spectroscopy for in vivo diagnosis of DMBA-induced hamster buccal pouch pre-cancers and cancers. AB - BACKGROUND: Our previous ex vivo study has shown that autofluorescence spectroscopy at 330-nm excitation can discriminate specimens of normal buccal pouch mucosa (normal), epithelial hyperkeratosis (hyperkeratosis), epithelial dysplasia (dysplasia), and squamous cell carcinoma (SCC) taken from DMBA-treated hamsters by using the method of partial least-squares discriminant analysis (PLSDA). METHODS: This study used a fiber optics-based fluorescence spectroscopy system to measure the autofluorescence spectra of 23 normal, 14 hyperkeratosis, 28 dysplasia, and 10 SCC samples in vivo. PLSDA with cross-validation was used to analyze the autofluorescence spectral data of all samples. RESULTS: We found that at 330-nm excitation, the autofluorescence spectra of all samples had two main peaks: one at 380 nm and the other at 460 nm. The hyperkeratosis samples had a higher 380-nm emission peak (EP) and a lower 460-nm EP than normal samples. On the contrary, the dysplasia samples had a lower 380-nm EP and a higher 460-nm EP than normal samples. Furthermore, the SCC samples had a much lower 380-nm EP and a much higher 460-nm EP than all other samples. To quantify the spectral changes during the progression of oral carcinogenesis, ratios of the area under the spectrum of 380 +/- 15 nm to that under the spectrum of 460 +/- 15 nm (denoted as A(380 +/- 15 nm)/A(460 +/- 15 nm)) for all samples were calculated. The mean ratio values of A(380 +/- 15 nm)/A(460 +/- 15 nm) decreased gradually from hyperkeratosis to normal, to dysplasia, and to SCC samples. Significant differences in this mean ratio were found between any two groups of normal, hyperkeratosis, dysplasia, and SCC samples. By choosing proper thresholds, PLSDA with cross-validation could provide an accurate identification rate of 86% for hyperkeratosis, of 87% for normal, and of 89% for dysplasia samples. In addition, by choosing a proper threshold, we could separate benign (normal and hyperkeratosis) from dysplasia or SCC tissues with a sensitivity of 92% and a specificity of 95%. CONCLUSION: Our results indicate that the autofluorescence spectroscopy technique is a useful diagnostic tool for in vivo diagnosis of oral pre-cancers and cancers in DMBA-induced hamster buccal pouch carcinogenesis model. PMID- 12558955 TI - Ultrastructural study of synovitis induced by trauma to the rat temporomandibular joint (TMJ). AB - BACKGROUND: Electron microscopy was used to examine the histologic effect of trauma on the rat temporomandibular joint synovial membrane. METHODS: Trauma to the TMJ in male Wister rats (100-200 g) was introduced through repeated forced condylar hypermobility. Ultrastructural observations were made 5 days and 6 weeks after the trauma. RESULTS: The early response of the synovial membrane was synovial hyperplasia, type A synovial cell loss, dilation of the r-ER in the type B synovial cells and fibrin deposition on the synovial surfaces. The late response included degeneration of synovial cells with swollen mitochondria and cell projections, and cell fragmentation. Large amount of fibrin deposition on opposing surface layers was also noticed. CONCLUSION: The type A cell loss and fibrin deposition followed by the occurrence of fibrinous materials at opposing surface layers of the synovial membrane suggest that traumatic synovitis causes synovial adhesions. PMID- 12558956 TI - Cytokine profile and supposed contribution to scarring in cicatricial pemphigoid. AB - BACKGROUND: The progressive scarring observed in cicatricial pemphigoid (CP) is still partially unexplained but recently the release of soluble fibrogenic factors by inflammatory infiltrating cells has been considered as pathogenically relevant. In the present study we evaluated the expression of mRNA for IL-4, IL 5, TGF-beta1, IFN-gamma in CP in comparison to bullous pemphigoid (BP) patients, investigating the role of cytokine profile as possible cause of the different disease evolution. METHODS: Biopsies from patients with oral (n = 10), preputial (n = 3) and cutaneous (n = 1) CP were studied by in situ hybridisation performing a new amplification system based on biotinyl-tyramide. As control, four patients affected by BP were also examined, together with healthy tissue from two CP and two BP patients, respectively. RESULTS: In CP IL-4 mRNA expression was present in 4 out of 14 cases analysed. IL-5 was detected in 12 CP biopsies. TGF-beta1 and IFN-gamma mRNAs were identified in 9 and 11 CP cases, respectively. In BP, IL-4 hybridisation signal could not be observed in any of the cases. By contrast IL-5, TGF-beta1 and IFN-gamma mRNA analyses were positive in all BP cases. Healthy specimens did not show any expression for IL-4, IL-5 and IFN-gamma, while a poor staining for TGF-beta was found in epithelium and subjunctional areas. CONCLUSIONS: Our results highlight the presence of a mixed cytokine pattern in the cellular infiltrate of both blistering diseases, with a corresponding increase of Th2-like activity in fully developed lesions, irrespective of the different sites involved. In addition, the constant presence of TGF-beta1 mRNA in the different lesional phases of CP, and its overlapping expression in BP suggest that the involvement of additional factors is responsible for the scarring course typical of CP. PMID- 12558957 TI - Thermal sensation and pain in oral lichen planus and lichenoid reaction. AB - BACKGROUND: Our previous findings in oral lichen planus (OLP) and lichenoid reactions (LR) raised the question whether the histopathological changes observed in sensory and autonomic innervation produce oral sensory disorders. METHODS: Spontaneous pain was assessed using the Visual Analogue Scale (VAS). Thermal pain thresholds were measured with a contact thermostimulator and mechanical pain thresholds with an electronic algometer. RESULTS: Patients with OLP reported a higher intensity of spontaneous pain than LR patients (P = 0.001). Even erosive LRs were relatively painless. No thermal or mechanical hyperalgesia was detected in oral lesions. Highest separate cold pain threshold was measured in lesions affected by intense Candida growth. CONCLUSION: Together with previous histological findings, the present data suggest that increased somatic innervation and sympathetic denervation do not promote clinical pain sensitivity or hyperalgesia in oral mucosa and that these sensory functions are not affected by OLP and LR. Candida growth may function as secondary irritant modulating the pain responses. PMID- 12558958 TI - Candida-induced stomatopyrosis and its relation to diabetes mellitus. AB - BACKGROUND: Glycaemic disorders and oral candidosis can be accompanied by burning mouth sensations. However, no clear relation between all three disorders is known. METHODS: Seventy-two native Upper-Austrians with burning mouth sensations were examined and smears for Candida estimation were taken from the spots where the sensations were felt. All patients with previously unknown diabetes mellitus (DM) were subjected to an oral glucose tolerance test (OGTT). Use of glucocorticoid-containing anti-asthmatic sprays and the body mass index (BMI) were determined. RESULTS: Of the examined non-inhalers of sprays, 52% had increased candidal density. A correlation between that increase and type 2 DM was found. The burning sensations in all patients with increased candidal density subsided completely after anti-mycotic therapy. CONCLUSION: The perception of burning sensations was hypothesised to occur via stimulation of the capsaicin (vanilloid) receptor by Candida metabolites. The Candida-induced stomatopyrosis should be regarded as a single symptom indicating (predisposition to or established) type 2 DM in non-inhalers of the concerned population. PMID- 12558959 TI - Successful treatment of invasive stomatitis due to Exophiala dermatitidis in a patient with acute myeloid leukemia. AB - BACKGROUND: Although the most common orofacial fungal infection in immunocompromised patients is candidosis, infections caused by virulent molds, such as Aspergillus spp. and Furarium spp. are being recognized with increasing frequency. We report a case of oral Exophiala infection in a 39-year-old woman with acute myeloid leukemia. METHODS: Clinical records of the patient were reviewed and the following additional information was collected: histological and microbiological evidence; identification of the causative organism; in vitro antifungal susceptibility. RESULTS: The patient developed a necrotic ulcer surrounded by a violaceous rim in the gingiva during neutropenia. Exophiala dermatitidis was identified as the causative organism by histopathological examination and culture, and finally confirmed by sequencing of the ribosomal DNA internal transcribed space domain. In vitro, amphotericin B was found to show strong activity against the Exophiala isolate while itraconazole showed less activity. The patient was successfully treated with parenteral amphotericin B and oral itraconazole in combination with surgical removal of the fungi focus. CONCLUSION: Local excision with adequate antifungal agents can be used to treat immunocompromised patients with Exophiala stomatitis, based on early diagnosis. PMID- 12558960 TI - A case of multiple AOT-like jawbone lesions in a young patient--a new odontogenic entity? AB - We assessed the immunohistochemical profile of an unusual case of multiple similarly looking tumors in the jawbone of a young patient. Histologically, the tumors exhibited features of adenomatoid odontogenic tumor (AOT) and adenomatoid dentinoma but showed no resemblance to any other defined odontogenic tumor entities. They expressed high amounts of cytokeratin (CK) 8 and 14 together with some Vimentin. A small rim of peripheral cells showed CK 5, 17, and 19 reactivity. Also, these lesions expressed some bcl-2 as well as p53 and Ki67. Histologically and immunohistochemically, the unusual multiple lesions differed in details from a simultaneously examined group of 24 classical AOT cases, suggesting that they may represent a hitherto less well-defined odontogenic tumor entity. PMID- 12558961 TI - Meal size and frequency affect neuronal plasticity and vulnerability to disease: cellular and molecular mechanisms. AB - Although all cells in the body require energy to survive and function properly, excessive calorie intake over long time periods can compromise cell function and promote disorders such as cardiovascular disease, type-2 diabetes and cancers. Accordingly, dietary restriction (DR; either caloric restriction or intermittent fasting, with maintained vitamin and mineral intake) can extend lifespan and can increase disease resistance. Recent studies have shown that DR can have profound effects on brain function and vulnerability to injury and disease. DR can protect neurons against degeneration in animal models of Alzheimer's, Parkinson's and Huntington's diseases and stroke. Moreover, DR can stimulate the production of new neurons from stem cells (neurogenesis) and can enhance synaptic plasticity, which may increase the ability of the brain to resist aging and restore function following injury. Interestingly, increasing the time interval between meals can have beneficial effects on the brain and overall health of mice that are independent of cumulative calorie intake. The beneficial effects of DR, particularly those of intermittent fasting, appear to be the result of a cellular stress response that stimulates the production of proteins that enhance neuronal plasticity and resistance to oxidative and metabolic insults; they include neurotrophic factors such as brain-derived neurotrophic factor (BDNF), protein chaperones such as heat-shock proteins, and mitochondrial uncoupling proteins. Some beneficial effects of DR can be achieved by administering hormones that suppress appetite (leptin and ciliary neurotrophic factor) or by supplementing the diet with 2-deoxy-d-glucose, which may act as a calorie restriction mimetic. The profound influences of the quantity and timing of food intake on neuronal function and vulnerability to disease have revealed novel molecular and cellular mechanisms whereby diet affects the nervous system, and are leading to novel preventative and therapeutic approaches for neurodegenerative disorders. PMID- 12558962 TI - Peripheral benzodiazepine receptors: on mice and human brain imaging. AB - There are numerous methods designed to monitor brain neuropathologies resulting from a wide arsenal of insults. Regardless of the cause of neuronal death, reactive glial cells always appear at and around the site of degeneration. These cells are distinguished by the exceptional abundance of peripheral benzodiazepine receptors, particularly compared with surrounding neurons. Measuring the binding of specific ligands to these peripheral benzodiazepine receptors offers a unique indirect marker for reliable damage assessment in the CNS and a faithful indicator for the accompanying cognitive deficits. PMID- 12558963 TI - Dose-dependent protective effect of selenium in rat model of Parkinson's disease: neurobehavioral and neurochemical evidences. AB - Normal cellular metabolism produces oxidants that are neutralized within cells by antioxidant enzymes and other antioxidants. An imbalance between oxidant and antioxidant has been postulated to lead the degeneration of dopaminergic neurons in Parkinson's disease. In this study, we examined whether selenium, an antioxidant, can prevent or slowdown neuronal injury in a 6-hydroxydopamine (6 OHDA) model of Parkinsonism. Rats were pre-treated with sodium selenite (0.1, 0.2 and 0.3 mg/kg body weight) for 7 days. On day 8, 2 micro L 6-OHDA (12.5 micro g in 0.2% ascorbic acid in normal saline) was infused in the right striatum. Two weeks after 6-OHDA infusion, rats were tested for neurobehavioral activity, and were killed after 3 weeks of 6-OHDA infusion for the estimation of glutathione peroxidase, glutathione-S-transferase, glutathione reductase, glutathione content, lipid peroxidation, and dopamine and its metabolites. Selenium was found to be successful in upregulating the antioxidant status and lowering the dopamine loss, and functional recovery returned close to the baseline dose-dependently. This study revealed that selenium, which is an essential part of our diet, may be helpful in slowing down the progression of neurodegeneration in parkinsonism. PMID- 12558964 TI - Synaptic activity induces signalling to CREB without increasing global levels of cAMP in hippocampal neurons. AB - Nuclear calcium signals associated with electrical activation of neurons can control the activity of the transcription factor cAMP-response element binding protein (CREB). Yet, cAMP is thought to be the key messenger that links synaptic activity to the regulation of CREB-mediated transcription. It is generally assumed that synaptic activity increases the intracellular levels of cAMP; this causes activation of the cAMP-dependent protein kinase (PKA) that regulates CREB mediated transcription either directly or through controlling nuclear signalling of the MAP kinases/extracellular signal-regulated kinases (ERK1/2) pathway. Here we show that, in hippocampal neurons, synaptic activity failed to increase global levels of cAMP that would be required for the cAMP-PKA system to induce nuclear events. Even near-continuous bursting of action potentials, giving rise to large nuclear calcium signals and robust CREB-dependent transcription, left global intracellular levels of cAMP unchanged. These results suggest that the cAMP-PKA system does not function as the transducer of synaptic signals to the nucleus. They indicate that the known inhibitory effects of blockers of PKA on gene expression and long-lasting plasticity triggered by calcium entry reflect a gating function of basal activity of PKA that renders neurons permissive for nuclear calcium-regulated, CREB/CBP-dependent gene expression. PMID- 12558965 TI - Histamine activates tyrosine hydroxylase in bovine adrenal chromaffin cells through a pathway that involves ERK1/2 but not p38 or JNK. AB - In bovine adrenal chromaffin cells (BACC) histamine promotes a rapid increase in the intracellular levels of Ca2+ together with the release of catecholamines and the phosphorylation of the catecholamine biosynthetic enzyme tyrosine hydroxylase (TH). In this study we investigated the role of the mitogen-activated protein kinases (MAPK) extracellular signal-regulated kinases (ERK1/2), stress activated protein kinase (p38) and Jun N-terminal kinases (JNK) on the histamine-induced activation and phosphorylation of TH. We found that in BACC histamine produced a rapid, long lasting and histamine type-1 (H1) receptor-dependent increase in the phosphorylation levels of ERK1/2, p38 and JNK which was accompanied by a H1 receptor-dependent increase in TH activity. This increase in TH activity was partially blocked by the MEK1/2 inhibitor U0126 but was unaffected by the p38 antagonist SB203580 or the JNK blocker JNKI1. To study the effect of MAPK inhibition on histamine-induced TH phosphorylation, we generated phospho-specific antibodies against the different phosphorylated forms of TH. Treatment with U0126 totally inhibited the histamine-induced phosphorylation of TH at Ser31, without affecting the phosphorylation of either Ser40 or Ser19. Neither SB203580 nor JNKI1 treatments produced any significant modification of the histamine-induced TH phosphorylation. Our data support the hypothesis that the up-regulation of the ERK1/2 pathway, but not that of p38 or JNK, promoted by histamine is involved in the phosphorylation of TH at Ser31 and that this phosphorylation event is required for the full activation of this enzyme. PMID- 12558966 TI - Level of haem oxygenase does not obligatorily reflect the sensitivity of PC12 cells to an oxidative shock induced by glutathione depletion. AB - In order to investigate the function of haem oxygenase in neuronal cell death or survival, we have determined in PC12 cells whether induction of haem oxygenase mRNA and protein or inhibition of haem oxygenase activity may be able to modulate the cell response to an oxidative stress. Inhibition of glutathione biosynthesis by buthionine sulfoximine (BSO) has indeed been demonstrated, in this cell line, to decrease the intracellular content of glutathione and to trigger a gradual and programmed cell death. Inhibition of haem oxygenase by zinc protoporphyrin IX, a potent inhibitor of this enzyme, or by a recently described peptidic inhibitor, induced a significant decrease in the toxicity of BSO. This protective action was not due to an alteration in the metabolism of glutathione and was still observed when the protecting agent was added several hours after BSO treatment. Induction of haem oxygenase-1 mRNA and protein by either haemin or pyrrolidine dithiocarbamate was associated with no protection or a significant reduction in the toxicity of BSO respectively. Our results indicate that induction of haem oxygenase-1 is not obligatorily associated with an improved resistance towards oxidative stress and suggest that a byproduct of haem degradation may also become detrimental. PMID- 12558967 TI - Cholesterol is necessary both for the toxic effect of Abeta peptides on vascular smooth muscle cells and for Abeta binding to vascular smooth muscle cell membranes. AB - Accumulation of beta amyloid (Abeta) in the brain is central to the pathogenesis of Alzheimer's disease. Abeta can bind to membrane lipids and this binding may have detrimental effects on cell function. In this study, surface plasmon resonance technology was used to study Abeta binding to membranes. Abeta peptides bound to synthetic lipid mixtures and to an intact plasma membrane preparation isolated from vascular smooth muscle cells. Abeta peptides were also toxic to vascular smooth muscle cells. There was a good correlation between the toxic effect of Abeta peptides and their membrane binding. 'Ageing' the Abeta peptides by incubation for 5 days increased the proportion of oligomeric species, and also increased toxicity and the amount of binding to lipids. The toxicities of various Abeta analogs correlated with their lipid binding. Significantly, binding was influenced by the concentration of cholesterol in the lipid mixture. Reduction of cholesterol in vascular smooth muscle cells not only reduced the binding of Abeta to purified plasma membrane preparations but also reduced Abeta toxicity. The results support the view that Abeta toxicity is a direct consequence of binding to lipids in the membrane. Reduction of membrane cholesterol using cholesterol lowering drugs may be of therapeutic benefit because it reduces Abeta-membrane binding. PMID- 12558968 TI - Tethering the N-terminus of the prion protein compromises the cellular response to oxidative stress. AB - The role of the N-terminal half of the prion protein (PrPC) in normal cellular function and pathology remains enigmatic. To investigate the biological role of the N-terminus of PrP, we examined the cellular properties of a construct of murine PrP, PrP-DA, in which the N-terminus is tethered to the membrane by an uncleaved signal peptide and which retains the glycosyl-phosphatidylinositol anchor. Human neuroblastoma SH-SY5Y cells expressing PrP-DA were more susceptible to hydrogen peroxide and copper induced toxicity than wtPrP expressing cells. The PrP-DA expressing cells had an increased level of intracellular free radicals and reduced levels of superoxide dismutase and glutathione peroxidase as compared to the wtPrP expressing cells. The membrane topology, cell surface location, lipid raft localisation, intracellular trafficking and copper-mediated endocytosis of PrP-DA were not significantly different from wtPrP. However, cells expressing PrP DA accumulated an N-terminal fragment that was resistant to proteinase K. The data presented here are consistent with the N-terminal region of PrPC having a role in the cellular response to oxidative stress, and that tethering this region of the protein to the membrane compromises this function through the accumulation of a protease-resistant N-terminal fragment, similar to that seen in some forms of human prion disease. PMID- 12558969 TI - Chronic systemic complex I inhibition induces a hypokinetic multisystem degeneration in rats. AB - In Parkinson's disease, nigral dopaminergic neurones degenerate, whereas post synaptic striatal target neurones are spared. In some atypical parkinsonian syndromes, both nigral and striatal neurones degenerate. Reduced activity of complex I of the mitochondrial respiratory chain has been implicated in both conditions, but it remains unclear if this affects the whole organism or only the degenerating brain structures. We therefore investigated the differential vulnerability of various brain structures to generalized complex I inhibition. Male Lewis rats infused with rotenone, a lipophilic complex I inhibitor [2.5 mg/kg/day intraveneously (i.v.) for 28 days], were compared with vehicle-infused controls. They showed reduced locomotor activity and loss of striatal dopaminergic fibres (54%), nigral dopaminergic neurones (28.5%), striatal serotoninergic fibres (34%), striatal DARPP-32-positive projection neurones (26.5%), striatal cholinergic interneurones (22.1%), cholinergic neurones in the pedunculopontine tegmental nucleus (23.7%) and noradrenergic neurones in the locus ceruleus (26.4%). Silver impregnation revealed pronounced degeneration in basal ganglia and brain stem nuclei, whereas the hippocampus, cerebellum and cerebral cortex were less affected. These data suggest that a generalized mitochondrial failure may be implicated in atypical parkinsonian syndromes but do not support the hypothesis that a generalized complex I inhibition results in the rather selective nigral lesion observed in Parkinson's disease. PMID- 12558970 TI - Rac and Cdc42-dependent regulation of c-Jun N-terminal kinases by the delta opioid receptor. AB - Heptahelical opioid receptors utilize Gi proteins to regulate a multitude of effectors including the classical adenylyl cyclases and the more recently discovered mitogen-activated protein kinases (MAPKs). The c-Jun NH2-terminal kinases (JNKs) belong to one of three subgroups of MAPKs. In NG108-15 neuroblastoma x glioma hybrid cells that endogenously express delta-opioid receptors, delta-agonist dose-dependently stimulated JNK activity in a pertussis toxin-sensitive manner. By using COS-7 cells transiently transfected with the cDNAs of delta-opioid receptor and hemagglutinin (HA)-tagged JNK, we delineated the signaling components involved in this pathway. Sequestration of Gbetagamma subunits by transducin suppressed the opioid-induced JNK activity. The possible involvement of the small GTPases was also examined. Expression of dominant negative mutants of Rac and Cdc42 blocked the opioid-induced JNK activation, and a partial inhibition was observed in the presence of the dominant negative mutant of Ras. In contrast, the dominant negative mutant of Rho did not affect the opioid-induced JNK activation. In addition, the receptor-mediated JNK activation was dependent on Src family tyrosine kinases, but independent of phosphatidylinositol-3 kinase and EGF receptor tyrosine kinases. Collectively, these results demonstrate functional regulation of JNK by the delta-opioid receptor, and this pathway requires Gbetagamma, Src kinases and the small GTPases Rac and Cdc42. PMID- 12558971 TI - Alterations in inducible 72-kDa heat shock protein and the chaperone cofactor BAG 1 in human brain after head injury. AB - The stress response in injured brain is well characterized after experimental ischemic and traumatic brain injury (TBI); however, the induction and regulation of the stress response in humans after TBI remains largely undefined. Accordingly, we examined injured brain tissue from adult patients (n = 8) that underwent emergent surgical decompression after TBI, for alterations in the inducible 72-kDa heat shock protein (Hsp70), the constitutive 73-kDa heat shock protein (Hsc70), and isoforms of the chaperone cofactor BAG-1. Control samples (n = 6) were obtained postmortem from patients dying of causes unrelated to CNS trauma. Western blot analysis showed that Hsp70, but not Hsc70, was increased in patients after TBI versus controls. Both Hsp70 and Hsc70 coimmunoprecipitated with the cofactor BAG-1. The 33 and 46, but not the 50-kDa BAG-1 isoforms were increased in patients after TBI versus controls. The ratio of the 46/33-kDa isoforms was increased in TBI versus controls, suggesting negative modulation of Hsp70/Hsc70 protein refolding activity in injured brain. These data implicate induction of the stress response and its modulation by the chaperone cofactor and Bcl-2 family member BAG-1, after TBI in humans. PMID- 12558972 TI - The 'glial' glutamate transporter, EAAT2 (Glt-1) accounts for high affinity glutamate uptake into adult rodent nerve endings. AB - The excitatory amino acid transporters (EAAT) removes neurotransmitters glutamate and aspartate from the synaptic cleft. Most CNS glutamate uptake is mediated by EAAT2 into glia, though nerve terminals show evidence for uptake, through an unknown transporter. Reverse-transcriptase PCR identified the expression of EAAT1, EAAT2, EAAT3 and EAAT4 mRNAs in primary cultures of mouse cortical or striatal neurones. We have used synaptosomes and glial plasmalemmal vesicles (GPV) from adult mouse and rat CNS to identify the nerve terminal transporter. Western blotting showed detectable levels of the transporters EAAT1 (GLAST) and EAAT2 (Glt-1) in both synaptosomes and GPVs. Uptake of [3H]D-aspartate or [3H]L glutamate into these preparations revealed sodium-dependent uptake in GPV and synaptosomes which was inhibited by a range of EAAT blockers: dihydrokainate, serine-o-sulfate, l-trans-2,4-pyrrolidine dicarboxylate (PDC) (+/-)-threo-3 methylglutamate and (2S,4R )-4-methylglutamate. The IC50 values found for these compounds suggested functional expression of the 'glial, transporter, EAAT2 in nerve terminals. Additionally blockade of the majority EAAT2 uptake sites with 100 micro m dihydrokainate, failed to unmask any functional non-EAAT2 uptake sites. The data presented in this study indicate that EAAT2 is the predominant nerve terminal glutamate transporter in the adult rodent CNS. PMID- 12558973 TI - Degraded myelin-associated glycoprotein (dMAG) formation from pure human brain myelin-associated glycoprotein (MAG) is not mediated by calpain or cathepsin L like activities. AB - The myelin-associated glycoprotein (MAG) is a transmembrane cell adhesion molecule participating in myelin formation and maintenance. Calcium-activated/ dependent proteolysis of myelin-associated glycoprotein by calpain and cathepsin L-like activities has already been detected in purified myelin fractions, producing a soluble fragment, called degraded (d)MAG, characterized by the loss of the transmembrane and cytoplasmic domains. Here, we demonstrate and analyze dMAG formation from pure human brain myelin-associated glycoprotein. The activity never exhibited the high rate previously reported in human myelin fractions. Degradation is time-, temperature-, buffer- and structure-dependent, is inhibited at 4 degrees C and by denaturation of the sample, and is mediated by a trans acting factor. There is no strict pH dependency of the proteolysis. Degradation was inhibited by excess aprotinin, but not by 1-10 micro g/mL aprotinin and was not eliminated by the use of an aprotinin-sepharose matrix during the purification. dMAG formation was not enhanced by calcium, nor inhibited by a wide variety of protease inhibitors, including specific calpain and cathepsin L inhibitors. Therefore, while cysteine proteases may be present in human myelin membrane fractions, they are not involved in dMAG formation from highly purified human brain myelin-associated glycoprotein preparations. PMID- 12558974 TI - The role of phosphatidylinositol 3-kinase in neural cell adhesion molecule mediated neuronal differentiation and survival. AB - The neural cell adhesion molecule, NCAM, is known to stimulate neurite outgrowth from primary neurones and PC12 cells presumably through signalling pathways involving the fibroblast growth factor receptor (FGFR), protein kinase A (PKA), protein kinase C (PKC), the Ras-mitogen activated protein kinase (MAPK) pathway and an increase in intracellular Ca2+ levels. Stimulation of neurones with the synthetic NCAM-ligand, C3, induces neurite outgrowth through signalling pathways similar to the pathways activated through physiological, homophilic NCAM stimulation. We present here data indicating that phosphatidylinositol 3-kinase (PI3K) is required for NCAM-mediated neurite outgrowth from PC12-E2 cells and from cerebellar and dopaminergic neurones in primary culture, and that the thr/ser kinase Akt/protein kinase B (PKB) is phosphorylated downstream of PI3K after stimulation with C3. Moreover, we present data indicating a survival promoting effect of NCAM-stimulation by C3 on cerebellar and dopaminergic neurones induced to undergo apoptosis. This protective effect of C3 included an inhibition of both DNA-fragmentation and caspase-3 activation. The survival promoting effect of NCAM-stimulation was also shown to be dependent on PI3K. PMID- 12558975 TI - The immunoglobulin-superfamily molecule basigin is a binding protein for oligomannosidic carbohydrates: an anti-idiotypic approach. AB - Recognition molecules that carry carbohydrate structures regulate cell interactions during development and play important roles in synaptic plasticity and regeneration in the adult. Glycans appear to be involved in these interactions. We have searched for binding proteins for oligomannosidic structures using the L3 antibody directed against high mannose-type glycans in an anti-idiotypic approach. A selected monoclonal anti-idiotype antibody was used for affinity chromatography and identified basigin as a binding protein from mouse brain detergent lysates. Basigin was found to bind to high mannose-carrying cell recognition molecules, such as myelin-associated glycoprotein, L1, the beta2 subunit of Na+/K+-ATPase and an oligomannosidic neoglycolipid. Furthermore, basigin was involved in outgrowth of astrocytic processes in vitro. A striking homology between the first immunoglobulin (Ig)-like domain of basigin and the fourth Ig-like domain of NCAM, previously shown to bind to oligomannosidic glycans, and the lectin domain of the mannose receptor confirms that basigin is an oligomannose binding lectin. To our knowledge this is the first report that anti-idiotypic antibodies can be used to identify binding partners for carbohydrates. PMID- 12558976 TI - Regulation of c-Jun N-terminal kinase, p38 kinase and AP-1 DNA binding in cultured brain neurons: roles in glutamate excitotoxicity and lithium neuroprotection. AB - In rat cerebellar granule cells, glutamate induced rapid activation of c-Jun N terminal kinase (JNK) and p38 kinase to phosphorylate c-Jun (at Ser63) and p53 (at Ser15), respectively, and a subsequent marked increase in activator protein-1 (AP-1) binding that preceded apoptotic death. These glutamate-induced effects and apoptosis could largely be prevented by long-term (7 days) pretreatment with 0.5 2 mm lithium, an antibipolar drug. Glutamate's actions could also be prevented by known blockers of this pathway, MK-801 (an NMDA receptor blocker), SB 203580 (a p38 kinase inhibitor) and curcumin (an AP-1 binding inhibitor). The concentration and time-dependent suppression of glutamate's effects by lithium and curcumin correlated well with their neuroprotective effects. These results suggest a prominent role of JNK and p38, as well as their downstream AP-1 binding activation and p53 phosphorylation in mediating glutamate excitotoxicity. Moreover, the neuroprotective effects of lithium are mediated, at least in part, by suppressing NMDA receptor-mediated activation of the mitogen-activated protein kinase pathway. PMID- 12558977 TI - Differential regulation of somatodendritic and nerve terminal dopamine release by serotonergic innervation of substantia nigra. AB - Nigrostriatal dopaminergic neurons release dopamine from dendrites in substantia nigra and axon terminals in striatum. The cellular mechanisms for somatodendritic and axonal dopamine release are similar, but somatodendritic and nerve terminal dopamine release may not always occur in parallel. The current studies used in vivo microdialysis to simultaneously measure changes in dendritic and nerve terminal dopamine efflux in substantia nigra and ipsilateral striatum respectively, following intranigral application of various drugs by reverse dialysis through the nigral probe. The serotonin releasers (+/-)-fenfluramine (100 micro m) and (+)-fenfluramine (100 micro m) significantly increased dendritic dopamine efflux without affecting extracellular dopamine in striatum. The non-selective serotonin receptor agonist 1-(m-chlorophenyl)-piperazine (100 micro m) elicited a similar pattern of dopamine release in substantia nigra and striatum. NMDA (33 micro m) produced an increase in nigral dopamine of a similar magnitude to mCPP or either fenfluramine drug. However, NMDA also induced a concurrent increase in striatal dopamine. The D2 agonist quinpirole (100 micro m) had a parallel inhibitory effect on dopamine release from dendritic and terminal sites as well. Taken together, these data suggest that serotonergic afferents to substantia nigra may evoke dendritic dopamine release through a mechanism that is uncoupled from the impulse-dependent control of nerve terminal dopamine release. PMID- 12558978 TI - Activation of capsaicin-sensitive primary sensory neurones induces anandamide production and release. AB - The inhibitory cannabinoid 1 receptor and the excitatory vanilloid receptor 1, both of which are responsive to the endogenous ligand anandamide, are co expressed on a subpopulation of primary sensory neurones. We report that activation of the cannabinoid 1 receptor/vanilloid receptor 1-co-expressing primary sensory neurones induces the production and release of anandamide. Application of capsaicin (3 nm-1 micro m) to cultured primary sensory neurones evoked calcitonin gene-related peptide release, which was significantly increased by the selective cannabinoid 1 receptor antagonist, SR141716A (200 nm). Mass spectrometric analyses of the extracellular solution revealed that exposure to capsaicin (10 nm or 100 nm) enhanced the anandamide concentration of the medium from less then 0.05 pmol/ micro L to more then 2 pmol/ micro L. Depolarization of the neurones with 50 mm KCl also enhanced the anandamide content of the buffer. Both the capsaicin- and KCl-induced anandamide release depended on extracellular Ca2+. Prolonged treatment of the cultures with capsaicin (10 micro m) reduced both the capsaicin- and KCl-induced anandamide release. These findings indicate that activation of capsaicin-sensitive primary sensory neurones evokes anandamide production and release, and that anandamide might be a key endogenous regulator of the excitability of these neurones. PMID- 12558979 TI - Arachidonic acid and anandamide have opposite modulatory actions at the glycine transporter, GLYT1a. AB - The GLYT1 subtypes of glycine transporter are expressed in glia surrounding excitatory synapses in the mammalian CNS and may regulate synaptic glycine concentrations required for activation of the NMDA subtypes of glutamate receptor. In this report we demonstrate that the rate of glycine transport by GLYT1 is inhibited by arachidonic acid. The cyclo-oxygenase and lipoxygenase inhibitors indomethacin and nordihydroguaiaretic acid, and the protein kinase C inhibitor staurosporine, had no effect on the extent of arachidonic acid inhibition of transport, which suggests that the inhibitory effects of arachidonic acid result from a direct interaction with the transporter. In contrast to arachidonic acid, its amide derivative, anandamide, and the more stable analogue R1-methanandamide stimulate glycine transport. This stimulation is unlikely to be a secondary effect of cannabinoid receptor stimulation because the cannabinoid receptor agonist WIN 55 212-2 had no effect on transport. We suggest that the stimulatory effects of anandamide on GLYT1 are due to a direct interaction with the transporter. PMID- 12558980 TI - Characterization of amyloid beta peptides from brain extracts of transgenic mice overexpressing the London mutant of human amyloid precursor protein. AB - Alzheimer's disease (AD) is marked by the presence of neurofibrillary tangles and amyloid plaques in the brain of patients. To study plaque formation, we report on further quantitative and qualitative analysis of human and mouse amyloid beta peptides (Abeta) from brain extracts of transgenic mice overexpressing the London mutant of human amyloid precursor protein (APP). Using enzyme-linked immunosorbant assays (ELISAs) specific for either human or rodent Abeta, we found that the peptides from both species aggregated to form plaques. The ratios of deposited Abeta1-42/1-40 were in the order of 2-3 for human and 8-9 for mouse peptides, indicating preferential deposition of Abeta42. We also determined the identity and relative levels of other Abeta variants present in protein extracts from soluble and insoluble brain fractions. This was done by combined immunoprecipitation and mass spectrometry (IP/MS). The most prominent peptides truncated either at the carboxyl- or the amino-terminus were Abeta1-38 and Abeta11-42, respectively, and the latter was strongly enriched in the extracts of deposited peptides. Taken together, our data indicate that plaques of APP-London transgenic mice consist of aggregates of multiple human and mouse Abeta variants, and the human variants that we identified were previously detected in brain extracts of AD patients. PMID- 12558982 TI - In situ imaging of specific binding of [3H]isatin in rat brain. AB - Isatin is an endogenous indole that influences a range of processes both in vivo and in vitro. It has a distinct and discontinuous distribution in the brain, as well as in other mammalian tissues and body fluids. However, the distribution of isatin binding sites in the brain is not known. Using a real-time beta-imager we have investigated the distribution of [3H]isatin-specific binding in rat brain sections. The highest labeling was found in hypothalamic nuclei and in the cortex, hippocampus, and cerebellum. Administration of the mechanism based monoamine oxidase inhibitor, pargyline, reduced but did not abolish the specific binding of [3H]isatin in the rat brain. The distribution became cortex, cerebellum, hypothalamus > hippocampus > brain stem > thalamus approximately striatum. PMID- 12558981 TI - Polyamine transport, accumulation, and release in brain. AB - Cycling of polyamines (spermine and spermidine) in the brain was examined by measuring polyamine transport in synaptic vesicles, synaptosomes and glial cells, and the release of spermine from hippocampal slices. It was found that membrane potential-dependent polyamine transport systems exist in synaptosomes and glial cells, and a proton gradient-dependent polyamine transport system exists in synaptic vesicles. The glial cell transporter had high affinities for both spermine and spermidine, whereas the transporters in synaptosomes and synaptic vesicles had a much higher affinity for spermine than for spermidine. Polyamine transport by synaptosomes was inhibited by putrescine, agmatine, histidine, and histamine. Transport by glial cells was also inhibited by these four compounds and additionally by norepinephrine. On the other hand, polyamine transport by synaptic vesicles was inhibited only by putrescine and histamine. These results suggest that the polyamine transporters present in glial cells, neurons, and synaptic vesicles each have different properties and are, presumably, different molecular entities. Spermine was found to be accumulated in synaptic vesicles and was released from rat hippocampal slices by depolarization using a high concentration of KCl. Polyamines, in particular spermine, may function as neuromodulators in the brain. PMID- 12558983 TI - Accumulation of 7S SNARE complexes in hippocampal synaptosomes from chronically kindled rats. AB - Kindling is a model of complex partial epilepsy wherein periodic application of an initially subconvulsive stimulus leads to first limbic and then generalized tonic-clonic seizures. Several laboratories have reported that augmented neurotransmitter release of l-glutamate is associated with the chronically kindled state. Neurotransmitter release requires membrane proteins called SNAREs, which form transmembrane complexes that participate in vesicle docking and are required for membrane fusion. We show here that kindling by entorhinal stimulation is associated with an accumulation of 7S SNARE complexes in the ipsilateral hippocampus. This increase of 7S SNARE complexes appears to begin early in the kindling process, achieves a peak with full kindling, and remains at this level for at least a month following cessation of further kindling stimuli. The increase is focal and permanently limited to the ipsilateral hippocampus despite progression to generalized electrographic and behavioral seizures. It is not seen in animals that receive electroconvulsive seizures, suggesting it is related to the kindling process itself. The duration and focality of increased 7S SNARE complexes with entorhinal kindling suggest that this is an altered molecular process associated with epileptogenesis. PMID- 12558984 TI - Association of Caspr/paranodin with tumour suppressor schwannomin/merlin and beta1 integrin in the central nervous system. AB - Caspr/paranodin is an essential neuronal component of paranodal axoglial junctions, associated with contactin/F3. Its short intracellular domain contains a conserved motif (GNP motif) capable of binding protein 4.1 domains [FERM domains (four point one, ezrin, radixin, moesin)]. Schwannomin/merlin is a tumour suppressor expressed in many cell types, including in neurons, the function and partners of which are still poorly characterized. We show that the FERM domain of schwannomin binds to the paranodin GNP motif in glutathione S-transferase (GST) pull down assays and in transfected COS-7 cells. The two proteins co immunoprecipitated in brain extracts. In addition, paranodin and schwannomin were associated with integrin beta1 in transfected cells and in brain homogenates. The presence of paranodin increased the association between integrin beta1 and schwannomin or its N-terminal domain, suggesting that the interactions between these proteins are interdependent. In jimpy mutant mice, which display a severe dysmyelination with deficient paranodal junctions, the interactions between paranodin, schwannomin and integrin beta1 were profoundly altered. Our results show that schwannomin and integrin beta1 can be associated with paranodin in the central nervous system. Since integrin beta1 and schwannomin do not appear to be enriched in paranodes they may be quantitatively minor partners of paranodin in these regions and/or be associated with paranodin at other locations. PMID- 12558985 TI - Recombinant human serotonin 5A receptors stably expressed in C6 glioma cells couple to multiple signal transduction pathways. AB - Human serotonin 5A (5-HT5A) receptors were stably expressed in undifferentiated C6 glioma. In 5-HT5A receptors-expressing cells, accumulation of cAMP by forskolin was inhibited by 5-HT as reported previously. Pertussis toxin-sensitive inhibition of ADP-ribosyl cyclase was also observed, indicating a decrease of cyclic ADP ribose, a potential intracellular second messenger mediating ryanodine sensitive Ca2+ mobilization. On the other hand, 5-HT-induced outward currents were observed using the patch-clamp technique in whole-cell configuration. The 5 HT-induced outward current was observed in 84% of the patched 5-HT5A receptor expressing cells and was concentration-dependent. The 5-HT-induced current was inhibited when intracellular K+ was replaced with Cs+ but was not significantly inhibited by typical K+ channel blockers. The 5-HT-induced current was significantly attenuated by 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) in the patch pipette. Depleting intracellular Ca2+ stores by application of caffeine or thapsigargin also blocked the 5-HT-induced current. Blocking G protein, the inositol triphosphate (IP3) receptor, or pretreatment with pertussis toxin, all inhibited the 5-HT-induced current. IP3 showed a transient increase after application of 5-HT in 5-HT5A receptor-expressing cells. It was concluded that in addition to the inhibition of cAMP accumulation and ADP ribosyl cyclase activity, 5-HT5A receptors regulate intracellular Ca2+ mobilization which is probably a result of the IP3-sensitive Ca2+ store. These multiple signal transduction systems may induce complex changes in the serotonergic system in brain function. PMID- 12558986 TI - Phosphorylation of cAMP response element-binding protein in cultured striatal neurons by metabotropic glutamate receptor subtype 5. AB - The metabotropic glutamate receptor subtype 5 (mGluR5) is densely expressed in striatal projection neurons. As a G protein-coupled receptor, mGluR5 may initiate an intracellular cascade that conveys extracellular signals to gene expression. This study investigated the possible role of mGluR5 in the inducible phosphorylation of a nuclear transcription factor, cAMP response element-binding protein (CREB), in primary cultures of striatal neurons from rat E19 embryos or neonatal day-1 pups. We found that selective activation of mGluR5 with a selective agonist (RS)-2-chloro-5-hydroxyphenylglycine (CHPG) induced a rapid and transient increase in phosphorylated CREB immunoreactivity in striatal neurons as analyzed by both immunocytochemistry and western blot. The increase in CREB phosphorylation was concentration-dependent, and seen in neurochemically identified GABAergic neurons. Pre-treatment with the mGluR5 antagonist 2-methyl-6 (phenylethynyl)pyridine hydrochloride (MPEP) blocked the CHPG phosphorylation of CREB. In contrast, the mGluR1 antagonist, 7-(hydroxyimino)cyclopropa[b]chromen-1a carboxylate ethyl ester (CPCCOet) did not alter CHPG-stimulated CREB phosphorylation. The mGluR5 antisense oligonucleotides, but not their controls, selectively reduced basal mGluR5 levels as well as CREB phosphorylation in response to CHPG addition. Lastly, using an immediate early gene c-fos as a reporter of inducible gene expression downstream to phosphorylated CREB, we found that CHPG induced a rapid and transient increase in c-fos mRNA levels in cultured neurons as revealed by quantitative in situ hybridization. The increase in c-fos was kinetically correlated well with the CREB phosphorylation and blocked by MPEP and the CREB antisense oligonucleotides. These results demonstrate a positive linkage from surface mGluR5 to CREB phosphorylation, which is able to facilitate immediate gene expression in striatal neurons. PMID- 12558987 TI - Mouse brain gene expression changes after acute and chronic amphetamine. AB - Gene expression changes are candidate mechanisms to contribute to long-term consequences of psychostimulant use. We use microarrays to examine the expression of 6340 genes in brains of mice killed 5 or 20 h following 14 day, twice-daily treatments with saline (SS), saline followed by a single 7.5 mg/kg amphetamine dose (SA), or repeated 7.5 mg/kg amphetamine doses (AA) that produce sensitization but no clear-cut neuronal toxicities. Arrays display robust hybridization for about 3600 transcripts. One hundred and seventeen of these expressed transcripts are candidate positives for drug-related changes, displaying > 1.8-fold differences from SS control values in whole brains of either SA or AA mice. Five transcripts reveal altered expression in both AA and SA mice. SA mostly enhances expression while AA treatments largely reduce expression. Fourteen SA and four AA changes in whole brain mRNA were replicated by > 1.8-fold changes in independent microarray assessments of either cerebral cortical or brainstem mRNAs, with more changes identified in frontal than in entorhinal/parietal cortical samples. About one-quarter of these changes persist in initial studies of mice killed 20 h after the last amphetamine injection. Each of these genes, including transcription factor, cellular regulatory, structural and other gene family members, are candidates to contribute to brain adaptations to psychostimulants. PMID- 12558988 TI - Specific interactions of neuronal focal adhesion kinase isoforms with Src kinases and amphiphysin. AB - Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that activates Src family kinases via SH2- and SH3-mediated interactions. Specific FAK isoforms (FAK+), responsive to depolarization and neurotransmitters, are enriched in neurons. We analyzed the interactions of endogenous FAK+ and recombinant FAK+ isoforms containing amino acid insertions (boxes 6,7,28) with an array of SH3 domains and the c-Src SH2/SH3 domain tandem. Endogenous FAK+ bound specifically to the SH3 domains of c-Src (but not n-Src), Fyn, Yes, phosphtidylinositol-3 kinase, amphiphysin II, amphiphysin I, phospholipase Cgamma and NH2-terminal Grb2. The inclusion of boxes 6,7 was associated with a significant decrease in the binding of FAK+ to the c-Src and Fyn SH3 domains, and a significant increase in the binding to the Src SH2 domain, as a consequence of the higher phosphorylation of Tyr-397. The novel interaction with the amphiphysin SH3 domain, involving the COOH-terminal proline-rich region of FAK, was confirmed by coimmunoprecipitation of the two proteins and a closely similar response to stimuli affecting the actin cytoskeleton. Moreover, an impairment of endocytosis was observed in synaptosomes after internalization of a proline-rich peptide corresponding to the site of interaction. The data account for the different subcellular distribution of FAK and Src kinases and the specific regulation of the transduction pathways linked to FAK activation in the brain and implicate FAK in the regulation of membrane trafficking in nerve terminals. PMID- 12558989 TI - Humanin rescues human cerebrovascular smooth muscle cells from Abeta-induced toxicity. AB - Cerebral amyloid beta-protein (Abeta) angiopathy (CAA) is a key pathological feature of Alzheimer's disease (AD) and related disorders. We have used human cerebrovascular smooth muscle (HCSM) cells as an in vitro model system to investigate the pathogenic mechanisms of the pathology of CAA. It was previously demonstrated that certain pathogenic forms of Abeta induce several pathologic responses in these cells, including fibril assembly at the cell surface, increased levels of Abeta precursor, degradation of HCSM cell alpha-actin and cell death. The recently discovered novel rescue factor humanin (HN) was shown to protect neuronal cells in culture from various AD-relevant insults including treatment with Abeta. In this report we investigated whether the HN peptide could rescue HCSM cells from Abeta-induced toxicity. We found that treatment of HCSM cells with 10 microm HN prevented pathogenic Abeta-induced HCSM cell death using a fluorescent cell viability assay, and degradation of HCSM alpha-actin was diminished shown by quantitative immunoblotting. However, Abeta deposition and fibril formation at the cell surface and increased levels of cell-associated AbetaPP were not affected by treatment with HN as demonstrated by a thioflavin T fluorescence assay and immunochemical methods, respectively. These results suggest that the protective effects of HN occur downstream of these cell surface molecular events. This is the first demonstration of a rescue factor for HCSM cells from Abeta-mediated cell death as well as being the first report to show that neuronal cells and HCSM cells may share a common downstream mechanism in the Abeta-induced cell death pathway. PMID- 12558990 TI - Down-regulation of Bcl-2 and Bcl-xL expression with bispecific antisense treatment in glioblastoma cell lines induce cell death. AB - The functions of the antiapoptotic proteins Bcl-2 and Bcl-xL were examined in glioblastoma cells. Expression of both Bcl-2 and Bcl-xL were found to be elevated in protein lysates from seven early passage cell lines derived from human glioblastoma tumors compared with non-neoplastic glial cells. Down-regulation of both bcl-2 and bcl-xL expression in glioblastoma cell lines U87 and NS008 with bcl-2/bcl-xL bispecific antisense oligonucleotide resulted in spontaneous cell death. The mechanism of cell death was partially caspase-dependent. Executioner caspase 6 and caspase 7, but not caspase 3, were involved in apoptosis induced by bcl-2/bcl-xL antisense treatment. Interestingly, western blots failed to demonstrate expression of caspase 3 in two of the seven glioblastoma cell lines examined. The data support the hypothesis that Bcl-2 and Bcl-xL are important in preventing cell death in glioblastoma cells. It also suggests that there are functional pathways capable of successful completion of caspase-dependent cell death in gliomas. These findings support a potential role of bcl-2/bcl-xL bispecifc antisense oligonucleotide therapy as a treatment strategy to enhance caspase-dependent cell death in patients with glioblastoma. PMID- 12558991 TI - Control of the Na+/Ca2+ exchanger 3 promoter by cyclic adenosine monophosphate and Ca2+ in differentiating neurons. AB - The human gene for member 3 of solute carrier family 8 (SLC8A3), encoding the Na+/Ca2+ exchanger isoform 3 (NCX3), was identified on chromosome 14q24.2. The minimal promoter region was predicted 250 bp upstream of exon 1. This was confirmed by luciferase reporter assays of pGL3-promoter constructs in transfected SH-SY5Y cells. The promoter activity was monitored during the differentiation of this cell line elicited by the sequential treatment with retinoic acid and brain-derived neurotrophic factor (BDNF). The activity was induced by cyclic AMP (cAMP) via the CRE (cAMP response element) and was stimulated by retinoic acid. The increase of intracellular Ca2+ induced by the partial depolarization of the plasma membrane with KCl down-regulated both the basal and the cAMP-stimulated transcription. The down-regulation of the latter may be mediated by the phosphorylation of the CRE-binding protein by a calmodulin dependent kinase (CaMKII). The exposure of cells to BDNF after treatment with retinoic acid rapidly induced promoter activity during the initial five hours and phosphorylation of CRE-binding protein during the first two hours. The promoter activity was further enhanced by cAMP, but became insensitive to Ca2+. In BDNF stimulated cells cAMP elevation caused the preferential phosphorylation of ATF1 instead of that of CRE-binding protein. PMID- 12558992 TI - Agonist-induced internalization of metabotropic glutamate receptor 1A: structural determinants for protein kinase C- and G protein-coupled receptor kinase-mediated internalization. AB - To investigate the role of the intracellular C-terminal tail of the rat metabotropic glutamate receptor 1a (mGlu1a) in receptor regulation, we constructed three C-terminal tail deletion mutants (Arg847stop, DM-I; Arg868stop, DM-II; Val893stop, DM-III). Quantification of glutamate-induced internalization provided by ELISA indicated that DM-III, like the wild-type mGlu1a, underwent rapid internalization whilst internalization of DM-I and DM-II was impaired. The selective inhibitor of protein kinase C (PKC), GF109203X, which significantly reduced glutamate-induced mGlu1a internalization, had no effect on the internalization of DM-I, DM-II, or DM-III. In addition activation by carbachol of endogenously expressed M1 muscarinic acetylcholine receptors, which induces PKC- and Ca2+-calmodulin-dependent protein kinase II-dependent internalization of mGlu1a, produced negligible internalization of the deletion mutants. Co expression of a dominant negative mutant form of G protein-coupled receptor kinase 2 (DNM-GRK2; Lys220Arg) significantly attenuated glutamate-induced internalization of mGlu1a and DM-III, whilst internalization of DM-I and DM-II was not significantly affected. The glutamate-induced internalization of mGlu1a and DM-III, but not of DM-I or DM-II, was inhibited by expression of DNM-arrestin [arrestin-2(319-418)]. In addition glutamate-induced rapid translocation of arrestin-2-Green Fluorescent Protein (arr-2-GFP) from cytosol to membrane was only observed in cells expressing mGlu1a or DM-III. Functionally, in cells expressing mGlu1a, glutamate-stimulated inositol phosphate accumulation was increased in the presence of PKC inhibition, but so too was that in cells expressing DM-II and DM-III. Together these results indicate that different PKC mechanisms regulate the desensitization and internalization of mGlu1a. Furthermore, PKC regulation of mGlu1a internalization requires the distal C terminus of the receptor (Ser894-Leu1199), whilst in contrast glutamate stimulated GRK- and arrestin-dependent regulation of this receptor depends on a region of 25 amino acids (Ser869-Val893) in the proximal C-terminal tail. PMID- 12558993 TI - Vitamin E blocks early events induced by 1-methyl-4-phenylpyridinium (MPP+) in cerebellar granule cells. AB - Exposure of cerebellar granule cells (CGCs) to 1-methyl-4-phenylpyridinium (MPP+) results in apoptotic cell death, which is markedly attenuated by co-treatment of CGCs with the radical scavenger vitamin E. Analysis of free radical production and mitochondrial transmembrane potential (DeltaPsim), using specific fluorescent probes, showed that MPP+ mediates early radical oxygen species (ROS) production without a loss of DeltaPsim. Exposure to MPP+ also produces an early increase in Bad dephosphorylation and translocation of Bax to the mitochondria. These events are accompanied by cytochrome c release from mitochondria to cytosol, which is followed by caspase 3 activation. Exposure of the neurons to vitamin E maintains Bad phosphorylation and attenuates Bax translocation, inhibiting cytochrome c release and caspase activation. MPP+-mediated cytochrome c release is also prevented by allopurinol, suggesting the participation of xanthine oxidase in the process. Our results indicate that free radicals play an active role in the MPP+ induced early events that culminate with cell death. PMID- 12558994 TI - Differences in multidrug resistance phenotype and matrix metalloproteinases activity between endothelial cells from normal brain and glioma. AB - Endothelial cells (ECs) are new targets for tumor therapy. In this work, we purified endothelial cells from intracerebral and subcutaneous experimental gliomas as well as from normal brain in order to define some of the phenotypical differences between angiogenic and quiescent brain vasculature. We show that the multidrug resistance genes encoding drug efflux pumps at the brain endothelium are expressed differently in normal and tumoral vasculature. We also show that ECs from gliomas present increased activity of gelatinase B (MMP9), key enzyme in the angiogenic process. Importantly, we observe a different phenotype between ECs in the intracerebral and subcutaneous models. Our results provide molecular evidence of phenotypic distinction between tumoral and normal brain vasculature and indicate that the EC phenotype depends on interactions both with tumor cells and also with the microenvironment. PMID- 12558995 TI - Martentoxin, a novel K+-channel-blocking peptide: purification, cDNA and genomic cloning, and electrophysiological and pharmacological characterization. AB - Martentoxin, a novel K+-channel-specific peptide has been purified and characterized from the venom of the East-Asian scorpion (Buthus martensi Karsch). The whole cDNA precursor sequence suggested that martentoxin was composed of 37 residues with a unique sequence compared with other scorpion neurotoxins. The genomic DNA of martentoxin showed an additional intron situated unexpectedly in the 5' UTR region, besides one located close to the C-terminal of the signal peptide. The patch-clamp recording found that martentoxin at the applied dose of 100 nm could strongly block large-conductance Ca2+-activated K+ (BKCa) currents in adrenal medulla chromaffin cells, and BKCa currents blocked by martentoxin could be fully recovered within 30 seconds after washing, which is at least 10 times faster than recovery after charybdotoxin. Meanwhile, a biosensor binding assay showed a fast association rate and a slow dissociation rate of martentoxin binding on rat brain synaptosomes. The binding of martentoxin on rat brain synaptosomes could be inhibited regularly by charybdotoxin, and gradually by toosendanin in a concentration-dependent manner, but not by either apamin or P03 from Buthus martensi. The results thus indicate that martentoxin is a new member in the family of K+-channel-blocking ligands. PMID- 12558996 TI - Manganese ethylene-bis-dithiocarbamate and selective dopaminergic neurodegeneration in rat: a link through mitochondrial dysfunction. AB - Manganese ethylene-bis-dithiocarbamate (Mn-EBDC) is the major active element of maneb, a pesticide linked to parkinsonism in certain individuals upon chronic exposure. Additionally, it has been shown to produce dopaminergic neurodegeneration in mice systemically coexposed to another pesticide, 1,1' dimethyl-4,4'-bipyridinium (paraquat). Here, we described a rat model in which selective dopaminergic neurodegeneration was produced by delivering Mn-EBDC directly to the lateral ventricles. After establishing this model, we tested whether Mn-EBDC provoked dopamine efflux in the striatum, a well-known phenomenon produced by the mitochondrial inhibitor 1-methyl-4-phenylpyridinium (MPP+), the active metabolite of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) that causes parkinsonism in humans, as well as in some animals. Finally, we investigated whether Mn-EBDC directly inhibited mitochondrial function in vitro using isolated brain mitochondria. Our data demonstrated that Mn-EBDC induced extensive striatal dopamine efflux that was comparable with that induced by MPP+, and that Mn-EBDC preferentially inhibited mitochondrial complex III. As mitochondrial dysfunction is pivotal in the pathogenesis of Parkinson's disease (PD), our results support the proposal that exposure to pesticides such as maneb, or other naturally occurring compounds that inhibit mitochondrial function, may contribute to PD development. PMID- 12558997 TI - Stabilization of the cyclin-dependent kinase 5 activator, p35, by paclitaxel decreases beta-amyloid toxicity in cortical neurons. AB - One hallmark of Alzheimer's disease (AD) is the formation of neurofibrillary tangles, aggregated paired helical filaments composed of hyperphosphorylated tau. Amyloid-beta (Abeta) induces tau hyperphosphorylation, decreases microtubule (MT) stability and induces neuronal death. MT stabilizing agents have been proposed as potential therapeutics that may minimize Abeta toxicity and here we report that paclitaxel (taxol) prevents cell death induced by Abeta peptides, inhibits Abeta induced activation of cyclin-dependent kinase 5 (cdk5) and decreases tau hyperphosphorylation. Taxol did not inhibit cdk5 directly but significantly blocked Abeta-induced calpain activation and decreased formation of the cdk5 activator, p25, from p35. Taxol specifically inhibited the Abeta-induced activation of the cytosolic cdk5-p25 complex, but not the membrane-associated cdk5-p35 complex. MT-stabilization was necessary for neuroprotection and inhibition of cdk5 but was not sufficient to prevent cell death induced by overexpression of p25. As taxol is not permeable to the blood-brain barrier, we assessed the potential of taxanes to attenuate Abeta toxicity in adult animals using a succinylated taxol analog (TX67) permeable to the blood-brain barrier. TX67, but not taxol, attenuated the magnitude of both basal and Abeta-induced cdk5 activation in acutely dissociated cortical cultures prepared from drug treated adult mice. These results suggest that MT-stabilizing agents may provide a therapeutic approach to decrease Abeta toxicity and neurofibrillary pathology in AD and other tauopathies. PMID- 12558998 TI - MsGC-II, a receptor guanylyl cyclase isolated from the CNS of Manduca sexta that is inhibited by calcium. AB - We describe the cloning of a receptor guanylyl cyclase, MsGC-II, from the CNS of the insect Manduca sexta. Sequence comparisons with other receptor guanylyl cyclases show that MsGC-II is most similar to a predicted guanylyl cyclase in the Drosophila genome and to vertebrate retinal guanylyl cyclases. When expressed in COS-7 cells, MsGC-II exhibited a low level of basal activity that was nearly abolished in the presence of 10 micro m calcium. Incubation with either a mammalian guanylyl cyclase-activating protein or Drosophila frequenin resulted in only mild stimulation of activity, whereas incubation of COS-7 cells expressing MsGC-II with a variety of Manduca tissue extracts failed to stimulate enzyme activity above basal levels. Analysis of the tissue distribution of MsGC-II revealed that it is nervous system specific. In the adult, MsGC-II is present in neurons in the optic lobes, antennal lobes and cellular cortex, but it is most highly expressed in subsets of intrinsic mushroom body neurons. Thus, MsGC-II appears to be a neural-specific receptor guanylyl cyclase whose activity may be regulated either directly or indirectly by calcium. PMID- 12558999 TI - Correlation of local changes in extracellular oxygen and pH that accompany dopaminergic terminal activity in the rat caudate-putamen. AB - Terminal activity causes an increase in local cerebral blood flow that can be quantified by measuring the accompanying increase in tissue oxygen. Alkaline pH changes can also follow neuronal activation. The purpose of these studies was to determine whether these changes in extracellular oxygen and pH correlate. Fast scan cyclic voltammetry was used to detect changes in dopamine, pH and oxygen levels simultaneously in the caudate-putamen after electrical stimulation of the substantia nigra in anesthetized rats. The biphasic increases in oxygen and pH followed similar time courses, and were delayed a few seconds from the immediate release and uptake of dopamine. The changes following administration of neurotransmitter receptor antagonists as well as agents that modulate blood flow were identical for oxygen and pH. Two distinct mechanisms were identified that give rise to the oxygen and pH changes: blood vessel dilatation caused by nitric oxide synthesis after muscarinic receptor activation and adenosine receptor activation. We conclude that changes in blood flow accompanying terminal activity cause alkaline pH shifts by the rapid removal of carbon dioxide, a component of the extracellular brain buffering system. PMID- 12559000 TI - Co-expression of vesicular glutamate transporters (VGLUT1 and VGLUT2) and their association with synaptic-like microvesicles in rat pinealocytes. AB - A vesicular glutamate transporter (VGLUT) is responsible for the accumulation of l-glutamate in synaptic vesicles in glutamatergic neurons. Two isoforms, VGLUT1 and VGLUT2, have been identified, which are complementarily expressed in these neurons. Mammalian pinealocytes, endocrine cells for melatonin, are also glutamatergic in nature, accumulate l-glutamate in synaptic-like microvesicles (SLMVs), and secrete it through exocytosis. Although the storage of l-glutamate in SLMVs is mediated through a VGLUT, the molecular nature of the transporter is less understood. We recently observed that VGLUT2 is expressed in pinealocytes. In the present study, we show that pinealocytes also express VGLUT1. RT-PCR and northern blot analyses indicated expression of the VGLUT1 gene in pineal gland. Western blotting with specific antibodies against VGLUT1 indicated the presence of VGLUT1 in pineal gland. Indirect immunofluorescence microscopy with a section of pineal gland and cultured cells indicated that VGLUT1 and VGLUT2 are co localized with process terminal regions of pinealocytes. Furthermore, immunoelectronmicroscopy as well as subcellular fractionation studies revealed that both VGLUT1 and VGLUT2 are specifically associated with SLMVs. These results indicate that both VGLUTs are responsible for storage of l-glutamate in SLMVs in pinealocytes. Pinealocytes are the first exception as to complementary expression of VGLUT1 and VGLUT2. PMID- 12559001 TI - Peptidase activities of the 20/26S proteasome and a novel protease in human brain. AB - Many neurodegenerative diseases are characterized by ubiquitin-positive protein aggregates or inclusion bodies. Ubiquitin-conjugated proteins are degraded by the 20/26S proteasome, and reduced proteasome peptidase activities in brain homogenates have been reported in pathologic lesions of Parkinson's and Alzheimer's diseases. However, it is unknown whether crude extracts of human brain contain other proteases having peptidase activities. We found a novel protease of molecular weight of approximately 105 kDa in normal human brain, which exhibited trypsin-like (T-L) and chymotrypsin-like (ChT-L) activities (corresponding to 52% and 21% of the total activities in crude extracts) but not peptidyl glutamyl peptide hydrolase activity. Both T-L and ChT-L activities of this protease were partially inhibited by proteasome inhibitors (MG132, lactacystin) and, in contrast to those of the proteasome, also by sodium dodecyl sulfate. A simple method to obtain a brain fraction specific to the 20/26S proteasome was developed. Our human brain data suggest that T-L and ChT-L activity levels of the proteasome reported previously may include those of the 105 kDa protease, an enzyme of as yet unknown biological significance, and that it is necessary to separate the proteasome from this protease to evaluate the actual status of the ubiquitin-proteasome system in neurodegenerative disorders. PMID- 12559002 TI - Apoptosis in cerebellar granule neurons is associated with reduced interaction between CREB-binding protein and NF-kappaB. AB - Cerebellar granule neurons undergo apoptosis when switched from medium containing depolarizing levels of potassium (high K+ medium, HK) to medium containing low K+ (LK). NF-kappaB, a ubiquitously expressed transcription factor, is involved in the survival-promoting effects of HK. However, neither the expression nor the intracellular localization of the five NF-kappaB proteins, or of IkappaB-alpha and IkappaB-beta, are altered in neurons primed to undergo apoptosis by LK, suggesting that uncommon mechanisms regulate NF-kappaB activity in granule neurons. In this study, we show that p65 interacts with the transcriptional co activator, CREB-binding protein (CBP), in healthy neurons. The decrease in NF kappaB transcriptional activity caused by LK treatment is accompanied by a reduction in the interaction between p65 and CBP, an alteration that is accompanied by hyperphosporylation of CBP. LK-induced CBP hyperphosphorylation can be mimicked by inhibitors of protein phosphatase (PP) 2A and PP2A-like phosphatases such as okadaic acid and cantharidin, which also causes a reduction in p65-CBP association. In addition, treatment with these inhibitors induces cell death. Treatment with high concentrations of the broad-spectrum kinase inhibitor staurosporine prevents LK-mediated CBP hyperphosphorylation and inhibits cell death. In vitro kinase assays using glutathione-S-transferase (GST)-CBP fusion proteins map the LK-regulated site of phosphorylation to a region spanning residues 1662-1840 of CBP. Our results are consistent with possibility that LK induced apoptosis is triggered by CBP hyperphosphorylation, an alteration that causes the dissociation of CBP and NF-kappaB. PMID- 12559003 TI - Protein kinase C-epsilon is involved in the adenosine-activated signal transduction pathway conferring protection against ischemia-reperfusion injury in primary rat neuronal cultures. AB - Adenosine activates a signal transduction pathway (STP) in the heart and the brain, conferring protection against ischemia-reperfusion insult. Activation of protein kinase C (PKC), probably mainly PKC-epsilon, has been demonstrated to be part of the heart STP, but its role in the neuronal pathway is less clear. Here, we provide proof for the participation of PKC-epsilon in the neuronal adenosine activated STP. Primary rat neuronal cultures were exposed to chemical ischemia by iodoacetate, followed by reperfusion. The cultured neurons were protected against this insult by activation of the adenosine mechanism, by N6-(R) phenylisopropyladenosine [R(-)-PIA], a specific A1 adenosine receptor agonist. Exposure of the cultures to bisindolylmaleimide I, a highly selective PKC inhibitor, abrogated the protection. The exposure of the cultures to R(-)-PIA was found to result in phosphorylation (activation) of PKC-epsilon. Furthermore, insertion into the cells of a specific peptide inhibitor of PKC-epsilon translocation (epsilonV1-2), also abrogated the protection conferred by R(-)-PIA. These results demonstrate that activation of PKC-epsilon is a vital step in the neuronal adenosine-activated STP. PMID- 12559004 TI - Effects of phosphatidylserine on p38 mitogen activated protein kinase, cyclic AMP responding element binding protein and nuclear factor-kappaB activation in resting and activated microglial cells. AB - In the last few years, the interaction between phosphatidylserine (PS), a phospholipid that becomes permanently exposed on the external cell surface in the early phases of apoptosis, and its specific receptor (PtdSerR) has emerged as a crucial event for the engulfing of apoptotic cells and for preventing the acquisition of pro-inflammatory functions by peripheral macrophages. Recently, we demonstrated that PtdSerR is expressed in microglial cultures purified from neonatal rat brain, and that PS-liposomes, used to mimic apoptotic cells, strongly reduce the lipopolysaccharide (LPS)-induced release of inflammatory mediators. Here, we show that in resting microglia, PS-liposomes induce cyclic AMP responding element binding protein (CREB) phosphorylation but do not activate nuclear factor-kappaB (NF-kappaB) and p38 mitogen-activated protein kinase (p38), in line with the non-inflammatory consequences of the recognition and removal of apoptotic cells by macrophages. In LPS-activated microglia, PS-liposomes did not affect NF-kappaB activation but inhibited the phosphorylation of p38 and delayed that of CREB. To our knowledge, this is the first biochemical evidence of the molecular signaling evoked by PS/PtdSerR interaction possibly related to repression of pro-inflammatory activities in microglial cells. PMID- 12559005 TI - Transfusion medicine illustrated. A surfeit of spherocytes. PMID- 12559006 TI - The blood supply chain, from donor to patient: a call for greater understanding leading to more effective strategies for managing the blood supply. PMID- 12559007 TI - Prevention of Yersinia enterocolitica, Pseudomonas fluorescens, and Pseudomonas putida outgrowth in deliberately inoculated blood by a novel pathogen-reduction process. AB - BACKGROUND: Yersinia enterocolitica, Pseudomonas fluorescens, and P. putida are responsible for a significant amount of the bacterial sepsis cases attributed to RBC transfusions. INACTINE is a pathogen-reduction process for RBCs, which consists of incubation of RBCs with PEN110 (proprietary compound) followed by automated washing of the RBCs. INACTINE is an electrophilic agent, which inactivates a wide range of viruses and WBCs by disruption of nucleic acid replication. The present study investigated the effect of the PEN110 process on Y. enterocolitica, P. fluorescens, and P. putida. STUDY DESIGN AND METHODS: Identical units of reduced CPD/ADSOL additive solution (AS-1) or CP2D/Nutricel additive solution (AS-3) RBCs were inoculated with 10 to 100 CFU per mL of either Y. enterocolitica, P. fluorescens, or P. putida. The control units were put on storage immediately after the bacterial spike. The test units were subjected to the PEN110 process and then stored. Sham control units were processed the same way as test units without addition of PEN110. Bacterial titer in all units was monitored during the 6-week storage period. RESULTS: No bacteria were detected in any of the RBC units (n = 9 for each microorganism) prepared using the PEN110 process throughout 6 weeks of storage. Substantial bacterial growth occurred in all control and in a majority of sham control units (11 out of 15 experiments). The bacterial inactivation by the INACTINE process was found to be equally effective in CPD/AS-1 and CP2D/AS-3 RBC units. CONCLUSION: The INACTINE process effectively prevented the outgrowth of Y. enterocolitica, P. fluorescens, and P. putida deliberately inoculated into WBC-reduced CPD/AS-1 and CP2D/AS-3 RBCs. The results demonstrated the crucial bactericidal role of PEN110 in the INACTINE process. PMID- 12559008 TI - Seven-day storage of apheresis platelets: report of an in vitro study. AB - BACKGROUND: The objective of this study was to determine the allowable platelet content limits for apheresis platelets stored for 7 days in a platelet storage bag (COBE ELP, Gambro BCT). METHODS: Apheresis platelets under controlled concentration and volume per bag were stored in plasma up to 8 days at 22 degrees C with horizontal agitation. Routinely evaluated in vitro platelet parameters were followed. Oxygen consumption was directly measured with a Clark-type electrode. All components were cultured in aerobic medium on Day 7. RESULTS: Twenty-four components were evaluated in storage configurations (median [range], 340 [110-402] mL, 1.32 [0.99-2.45] x 10(6) platelets/microL, and 4.8 [1.4-5.9] x 10(11) platelets/bag). No bacterial contamination was detected. One component had a pH value at 22 degrees C of below 6.0 before Day 5 with attendant loss of all other in vitro function measures. The pH value at 22 degrees C was maintained above 6.2 for the remaining 23 components. A pH value of greater than 7.4 was observed at some point in storage for 13 of 23 units, although platelet function or activation was not adversely affected. Aerobic metabolic function was maintained over 7 days with O2 consumption of 321 micromol per hour per 10(12) platelets on Day 7. CONCLUSION: Although a continuing decline of platelet in vitro characteristics can be observed for storage beyond 5 days, apheresis platelets in plasma stored 100 to 400 mL per bag, 1.0 x 10(6) to 2.5 x 10(6) platelets per microL, and a maximum of 5.1 x 10(11) platelets per bag maintained in vitro platelet characteristics over 7 days of storage. PMID- 12559009 TI - WBC reduction reduces storage-associated RBC adhesion to human vascular endothelial cells under conditions of continuous flow in vitro. AB - BACKGROUND: The effects of storage duration, WBC reduction, and irradiation on RBC adherence to vascular endothelia are unknown and are investigated under conditions of continuous flow. STUDY DESIGN AND METHODS: Thirty-two RBC units were collected and divided into three groups, non-WBC-reduced (NWR), WBC-reduced (WR), and irradiated-WBC-reduced. Aliquots of RBCs were removed on Days 1, 15, and 28 of storage for analysis. The RBC suspensions were then perfused at a 1.5 percent Hct in a protein-poor medium under conditions of continuous flow over human umbilical vein endothelial cell monolayers. On each slide, 25 randomly chosen sites were videorecorded over 10 minutes, and the number of RBCs adherent to the endothelial cell monolayer was counted. RESULTS: NWR RBCs stored for 28 days demonstrated a greater degree of adherence to endothelial cells compared to Days 1 and 15 (p < 0.03). The WR group had significantly fewer adherent RBCs than the NWR group on day 28 (p < 0.01). Irradiation had no effect on RBC adherence. CONCLUSION: Prolonged storage of NWR RBCs increases RBC adherence to endothelial cells in vitro. WBC reduction before storage abrogates the effect of storage duration on increased adhesion. Studies to assess whether an in vivo effect occurs are required. PMID- 12559010 TI - Stability after thawing of RBCs frozen with the high- and low-glycerol method. AB - BACKGROUND: RBCs can be frozen with either the high-glycerol method (HGM) or the low-glycerol method (LGM). To date, the use of frozen RBCs is hampered by a 24 hour outdating period after thawing. A closed washing system (ACP 215) may solve this problem. STUDY DESIGN AND METHODS: We compared the effects of high- (40%) and low-glycerol (19%) concentration, with and without freezing (at -80 degrees C for HGM, -196 degrees C for LGM) on the in vitro quality of RBCs after deglycerolization with the closed washing system and during storage at 4 degrees C in SAGM after thawing. RESULTS: Glycerol treatment by itself induced hemolysis during processing, which was more pronounced in HGM cells. The freeze-thaw-wash process decreased the stability of RBCs, particularly in LGM cells during storage after thawing. In contrast to LGM cells, in HGM cells no additional effect of freeze or thaw on stability of washed cells was seen during the first week of storage after thawing. Changes in osmotic resistance and cellular metabolism could not explain the observed differences in RBC stability. CONCLUSION: The closed washing system is able to process both high- and low-glycerol-treated RBCs. Stability after washing during cold storage in SAGM, as measured by hemolysis, is better for HGM cells as compared to LGM cells. PMID- 12559011 TI - Blood-donor perceptions of health history screening with a computer-assisted self administered interview. AB - BACKGROUND: Computer-assisted self-interviewing (CASI) has been shown to promote disclosures of sensitive information. Using CASI to screen blood-donor candidates might encourage reports of deferrable risks without discouraging eligible donors. STUDY DESIGN AND METHODS: In 1998, an anonymous mail survey was sent to donors from eight blood centers. The relationship of donor demographics, donation history, and infectious risks (HIV test-seeking behavior, unreported deferrable risk, or reactive donor-screening test result) on attitudes toward CASI were assessed. RESULTS: Of 92,581 blood donors surveyed, 52,650 (57%) responded; of these, 4713 (7%) had an infectious risk. Among donors with risks, 29 percent felt they would be encouraged to reveal personal data, and 7 percent reported they would be discouraged from donating by CASI. Young, minority, female, and first time donors were the most likely to report that CASI would encourage personal disclosures. Among donors without risks, 5 percent felt CASI would discourage them from donating. CONCLUSIONS: Although most donors felt their donation practices would not be influenced, CASI might reduce the proportion of donors with infectious risks by fostering personal disclosures and self-deferrals. The potential for CASI to improve donor screening and increase appropriate self deferrals should be balanced against the possible loss of reluctant safe donors. PMID- 12559012 TI - Absence of D- alloimmunization in AIDS patients receiving D-mismatched RBCs. AB - BACKGROUND: More than 80 percent of D- patients who receive D+ blood become alloimmunized to the D antigen. Anemia occurs in most AIDS patients at some point in the disease. D- patients with AIDS may require blood transfusion and, during times of blood shortage, may receive D+ RBCs. They would be expected to become alloimmunized to the d antigen. STUDY DESIGN AND METHODS: The records of the transfusion service between January 1996 and July 2000 were reviewed for D- patients who received D+ blood. IATs were performed before the initial transfusion and subsequently when the patient required further RBC transfusion. RESULTS: Eight D- AIDS patients who received multiple transfusions (three women and five men; age range, 31-44 years; mean, 44 years) who received between 2 and 11 units (mean, 6.25) of D+ RBCs were identified. Antibody screens were performed at 8 to 65 weeks after transfusion. It was found that none of the eight D- AIDS patients developed anti-D. ABO antibodies were found as expected. During the same period, it was found that six D- patients admitted with other diagnoses who received 1 to 9 units of D+ RBCs, all developed anti-D within 7 to 19 weeks of transfusion. CONCLUSION: Patients with AIDS may not form alloantibodies to the D antigen. This may be attributable to their immunodepressed state, particularly to the decrease in CD4+ T lymphocytes. Therefore, during blood shortages, transfusion of D+ blood to D- AIDS patients may be without any subsequent consequence. PMID- 12559013 TI - TRALI: correlation of antigen-antibody and monocyte activation in donor-recipient pairs. AB - BACKGROUND: TRALI may be a severe reaction associated with transfusion of plasma containing blood components. TRALI has usually been associated with antibodies against granulocytes and HLA class I antigens, but more recently with antibodies against HLA class II and monocytes. TRALI cases were investigated to determine correlation between antigen and antibody. Additionally, activation of monocytes by TRALI serums was studied. STUDY DESIGN AND METHODS: Sixteen cases of TRALI were investigated. All patients were typed for HLA antigens. Implicated donors were screened for HLA antigens and antibodies against granulocytes and monocytes. In 6 cases, recipient monocyte activation was measured in vitro after incubation with TRALI and control serums. In four cases, monocyte activation was measured after incubation of TRALI serums against a panel of monocytes of known HLA antigen type. RESULTS: In 14 of the 16 cases (87.5%), antigen-antibody correlation was identified. TRALI monocytes, incubated with implicated TRALI serum (n = 6), expressed significantly greater cytokine and tissue factor (p < 0.05, repeated-measures ANOVA) than controls. Panel monocytes incubated with TRALI serum showed increased expression of cytokine and/or tissue factor when corresponding antigen was present. CONCLUSION: In most cases of TRALI, a correlation between antigen and antibody can be identified. Activation of monocytes and their subsequent release of cytokines may play a role in the pathogenesis of TRALI. PMID- 12559014 TI - Investigations into the role of anti-HLA class II antibodies in TRALI. AB - BACKGROUND: TRALI is thought to be triggered by recipient-specific anti-HLA class I or antibodies against neutrophils in donor plasma. Recently, anti-HLA class II have also been implicated. The prevalence of anti-HLA class II was investigated in normal volunteer platelet donors and in two nonfatal TRALI cases utilizing a flow-based assay. Potential target antigens also were investigated. STUDY DESIGN AND METHODS: Commercial flow cytometry-based assays (FlowPRA, One Lambda, Inc.) for anti-HLA class I and II were compared with standard lymphocytotoxicity tests. Subsequently, 151 volunteer platelet donors and two clinical cases of TRALI were screened with FlowPRA. Immunohistochemical studies were performed on lung tissue from a surgical case, an autopsy case, and a fatal TRALI case. RESULTS: The FlowPRA assays showed moderate concordance for anti-HLA class I (kappa = 0.448) and good concordance for class II antibodies (kappa = 0.801), when compared to standard lymphocytotoxicity assays. Ten and 9 percent of female platelet donors were positive for anti-HLA class I and class II, respectively. Two nonfatal cases of TRALI showed both anti-HLA class I and anti-HLA class II. Immunohistochemical analysis of a TRALI case revealed granulocyte aggregation in alveolar capillaries with activated vascular endothelial cells. HLA class II antigen expression was not present on vascular endothelium or intravascular WBCs; however, strong expression was seen on alveolar macrophages. CONCLUSION: FlowPRA assays often detect anti-HLA class I not detected by conventional lymphocytotoxicity assays. These assays reveal anti-HLA class II in normal female donor plasma and in sera implicated in TRALI. Immunohistochemical studies failed to reveal endothelial or intravascular-WBC HLA class II antigen expression in lung tissue derived from TRALI cases or controls, but demonstrated HLA class II expression on pulmonary macrophages. PMID- 12559015 TI - Health and economic impact of posttransfusion hepatitis B and cost-effectiveness analysis of expanded HBV testing protocols of blood donors: a study focused on the European Union. AB - BACKGROUND: Residual risk of posttransfusion hepatitis B (PT-HB) may be reduced through implementation of HBV NAT or the new, enhanced-sensitivity HBsAg assays in routine donor testing. However, there are some doubts about the cost effectiveness of these new safety measures, because hepatitis B acquired in adulthood is not regarded as a severe disease in western countries. STUDY DESIGN AND METHODS: A computer model was designed to estimate the health outcomes and associated costs of patients with PT-HB. Results from this model and estimations of the residual risk of HBV transmission, the risk reduction yielded by the new assays, and their cost were used to calculate the cost-effectiveness of including the new HBsAg assays or single-sample HBV NAT in the routine screening of blood donors. RESULTS: The model predicts that 0.97 percent of patients with PT-HB die of liver disease (54% of them due to fulminant hepatitis). The mean loss of life expectancy was 0.178 years per patient, and the present value of the lifetime costs of treating PT-HB was 4160 euros per patient. Single-donor HBV NAT or the new HBsAg assays would increase the life expectancy of blood recipients by 16 (95% CI, 8-40) or 14 (95% CI, 7-28) years, respectively, per every 10 million donations tested. The projected cost per life-year gained was 0.79 (95% CI, 0.15 1.85) million euros for the enhanced-sensitivity HBsAg assays and 5.8 (95% CI, 1.9-13.1) million euros for single-donation HBV NAT, both compared with current HBsAg assays. If single-donation HBV NAT is compared with the new HBsAg assays, its cost- effectiveness ratio increases to 53 (95% CI, 16-127) million euros. CONCLUSION: PT-HB has few health or economic repercussions. Single-donation HBV NAT would provide a small health benefit at a very high cost. Instead, in some circumstances, the cost-effectiveness of enhanced-sensitivity HBsAg assays would be within acceptable ranges for new public health interventions. PMID- 12559016 TI - Impact of changes in viral marker screening assays. AB - BACKGROUND: Monitoring the performance of routinely used infectious disease serologic tests is necessary to evaluate their effectiveness in identifying true positive units and erroneously disqualifying safe blood donors. METHODS: With two large screening test data sets collected between 1991 and 1998 and between 1997 and 2000, the impact of changes in screening assays for HIV, HCV, and HBsAg was analyzed with regard to the prevalence of confirmed-positive, indeterminate, and confirmed-negative results and the deferral of donors with an indeterminate or negative results (donor loss). RESULTS: The prevalence of indeterminate results and donors loss increased significantly in the 6 months after introduction of an HIV-1/2 EIA. A second-generation HCV EIA increased the detection of confirmed positive donations in repeat donors (p < 0.001) and increased the prevalence of indeterminate donations. Implementation of a third-generation HCV EIA resulted in a significant decrease in indeterminate results in first-time donors. Nonspecific test results increased when HBsAg test kits from a different manufacturer were introduced or different lots of HIV antibody screening test kits from the same manufacturer were used. CONCLUSION: Introduction of newly licensed versions of assays, switching kit manufacturers, and lot-to-lot variations have an impact on rates of deferrals of safe donors as well as sensitivity of routine screening. Before considering changes in screening tests, blood centers should be aware of, and evaluate, the potential impact on donor loss. PMID- 12559017 TI - Evaluation of a transcription-mediated amplification-based HCV and HIV-1 RNA duplex assay for screening individual blood donations: a comparison with a minipool testing system. AB - BACKGROUND: NAT was introduced for HCV RNA in 1999 to screen blood donations and improve the safety of the blood supply. STUDY DESIGN AND METHODS: The performance of a NAT multiplex for HCV and HIV-1 RNA based on transcription-mediated amplification (TMA) was assessed with various sensitivity panels and by screening 50,000 serologically unscreened, first-time donor plasma samples. Results were compared with a routine NAT screening for HCV RNA by RT-PCR in pools of 96 plasma samples. RESULTS: The TMA multiplex 95 percent sensitivity ranged between 22 and 54 IU per mL for HIV-1 and 15 and 20 IU per mL for HCV RNA. The rate of test failure was 8.6 percent but decreased to 4.7 percent when results of two critical periods of equipment malfunction were excluded. Test failure was related to human error, minute control contamination, and insufficient mixing of reagents at the extraction stage. All 31 repeatedly reactive samples (0.06%) were seropositive for HCV (29) or HIV-1 (2) and contained RNA detectable by discriminatory TMA and confirmatory RT-PCR, indicating 100 percent specificity. A direct comparison of TMA in individual samples and RT-PCR in plasma pools was possible on 27 HCV RNA containing samples. Twenty-six samples were detected in plasma pools; the lack of detection of 1 sample was due to an identification error at the pooling stage. CONCLUSION: The HCV and HIV-1 multiplex NAT had high specificity and sensitivity. PMID- 12559018 TI - Improved efficiency of national HIV, HCV, and HTLV antibody testing algorithms based on sequential screening immunoassays. AB - BACKGROUND: Traditional strategies for clarifying the antibody status of donors giving repeatedly reactive (RR) results on primary screening immunoassays (IA1) have usually involved direct testing by immunoblot. However, such strategies can generate nonspecific in determinate results. The aim of this report is to present the results of an alternative strategy based on the use of sequential immunoassays (SI) before immunoblot testing. STUDY DESIGN AND METHODS: The efficiency of traditional and SI strategies was compared in terms of the number of IA1 RR samples requiring immunoblot testing and the percentage of immunoblot tests giving indeterminate results. In addition, the biologic false- reactive overlap between the PRISM assays selected as IA1 and candidate secondary screening immuno- assays (IA2) was calculated to determine the most efficient IA1/IA2 combinations. RESULTS: There was a significant decrease in the proportion of IA1 RR samples requiring immunoblot testing under the SI strategy when compared with existing site-specific strategies for HIV (0.49 vs. 0.08, p < 0.05), HCV (0.85 vs. 0.42, p < 0.05), and HTLV (0.69 vs. 0.05, p < 0.05) algorithms. In addition, there was a significant decrease in the percentage of immunoblot tests giving indeterminate results for HIV and HTLV under the SI strategy. However, there was no significant difference in the proportion of confirmed positive results for HIV, HCV, or HTLV before and after national SI algorithm implementation. For the anti-HIV IA2s, there was considerable variation of biologic false-reactive overlap with the PRISM HIV O plus chemiluminescent immunoassay (range, 1.6-15.6%). CONCLUSIONS: The results presented in this report demonstrate that the sequential use of screening immunoassays before immunoblot testing can significantly reduce both the number of immunoblot tests and proportion of indeterminate results, without impacting sensitivity, thereby improving algorithm efficiency and simplifying donor management. PMID- 12559019 TI - Heart rate increases linearly in response to acute isovolemic anemia. AB - BACKGROUND: The cardiovascular response to acute isovolemic anemia in humans is thought to differ from that of other species. Studies of anesthetized humans have found either no change or a decreased heart rate. A previous study showed that in 32 healthy unmedicated humans, heart rate increased during acute isovolemic anemia. The hypothesis that heart rate in humans increases in response to acute isovolemic anemia and that the increase is affected by gender was tested. STUDY DESIGN AND METHODS: Acute isovolemic anemia to a Hb concentration of approximately 5 g per dL in 95 unmedicated healthy humans was produced by simultaneous withdrawal of blood and IV replacement with 5-percent HSA and autologous platelet-rich plasma. The relationship between heart rate and Hb concentration was examined using a mixed-effects linear regression model that allowed each person to have a fitted line with its own slope and intercept. Cubic and quadratic terms were added to determine if these improved the goodness of fit. The effect of gender was tested by including it and its interactions with Hb in the mixed model. RESULTS: The relationship between heart rate and Hb concentration was linear (p < 0.001) and consistent among the population studied: heart rate = 116.0-4.0 [Hb] (slope 95% CI: -4.2 to -3.8 beats/min/g Hb). Adding a cubic or quadratic term did not significantly improve the goodness of fit of the mathematical expression to the data, confirming the linear nature of the relationship between heart rate and Hb concentration. For women, the slope of the heart rate response was significantly greater than it was for males (difference +/- SE: 0.70 +/- 0.23, p < 0.005). CONCLUSION: In 95 unmedicated, healthy humans, heart rate was a linear function of Hb during acute isovolemic anemia. Females had a significantly greater slope of increase in heart rate with decreasing Hb concentration than did males. The relationship is consistent among individuals, is similar to that reported for conscious dogs, and differs from that found previously in anesthetized humans. PMID- 12559020 TI - Long-term production of pre-existing alloantibodies to E and c after allogenic BMT in a patient with aplastic anemia resulting in delayed hemolytic anemia. AB - BACKGROUND: Delayed hemolysis mediated by long-term production of pre-existing recipient-derived antibodies directed against donor RBC antigens after allogenic BMT is an unusual complication of hematopoietic transplantation. CASE REPORT: A 26-year-old man with aplastic anemia had pre-existing alloantibodies to E and c. He received BMT from a donor, whose Rh phenotype was E+, c+. From about 1 month after the transplant, he showed mild hemolysis due to the antibodies. RESULTS: The patient was typed as group B, CCDee and had anti-E and c alloantibodies before BMT. The donor was typed as group O, ccDEE. Although MNCs from the donor marrow were infused into the patient, DAT became positive on Day 21, and the patient-origin antibodies remained detectable by both DAT and IAT even 20 months after BMT. However, immunomagnetically isolated peripheral circulating B cells were 100 percent donor origin. The patient received prednisolone from Day 221, and thereafter, the signs of hemolysis disappeared. CONCLUSION: It is likely that the long-term production of alloantibodies is due to the existence of long-lived recipient plasma cells, which survive the conditioning regimen. This case suggests that patients with pre- existing alloantibodies that do not belong to the ABO system should be carefully followed up after BMT. PMID- 12559021 TI - Modulations of anti-D affinity following promiscuous binding of the heavy chain with naive light chains. AB - BACKGROUND: It is generally accepted that the antibody heavy (H) chain is more important than the light (L) chain for determining antigen specificity. In the case of anti-D, the predominant role of H chains in antigen binding is well recognized, but much less is known about the function of L chains. In this work, the contribution of L chains from non-D-immunized donors to the specificity and reactivity of anti-Ds was studied with L-chain shuffling. STUDY DESIGN AND METHODS: A kappa L chain library was recombined with the H chain of the 43F10 anti-D in a phagemid vector system (pComb3H, Scripps Institute). D-specific F(ab) phages were selected by panning on RBCs. Soluble F(ab)s were prepared, and their reactivity was assessed by RBC agglutination. The nucleotide and amino acid sequences of the variable region of the L chains were analyzed. RESULTS: The L chains of the six D-specific 43F10 F(ab) clones studied used five different germline genes from three Vkappa families and three different Jkappa segments. The L chains were all cationic with isoelectric points ranging from 8.1 to 10.2. CONCLUSION: The 43F10 anti-D H chain could bind promiscuously to a diversity of L chains from non-D-immunized donors without losing the D-antigen specificity. Relationships between the anti-D affinity and the cationic charge of the L chain as well as with the presence of an arginine residue in the L-chain complementarity-determining region 1 were observed. PMID- 12559022 TI - Use of percutaneous radial artery catheter for peripheral blood progenitor cell collection in pediatric patients. AB - BACKGROUND: Leukapheresis procedures require adequate flow rates, which in children may frequently involve invasive vascular access placement. STUDY DESIGN AND METHODS: A minimally invasive peripheral radial artery catheter was used for drawing blood in 85 leukapheresis procedures performed in 33 pediatric patients. Blood return to the patients was provided by either a central Broviac-type catheter or a peripheral venous access. The patients' age range was 1 to 18 years (median, 9.5) and the weight range was 9 to 73 kg (median, 29 kg). Vasocan Braunule Luer Lock IV cannulas (22 gauge in 78 and 20 gauge in 7) were placed percutaneously under local anesthesia, and in 8 patients, catheter placement was carried out during general anesthesia for other procedures. A continuous flow cell separator was used in all cases (Fresenius AS104 in 23 and AS204 in 62). RESULTS: Flow rates ranged from 18 to 45 mL per minute, the mean number of total blood volumes processed was 2.07 (range, 0.51-2.51), and the mean duration of the procedures was 150 minutes (range, 90-260). The 22-gauge cannulas provided adequate flow rates independently of patient age and weight. No significant thrombotic, embolic, hemorrhagic, ischemic, or infectious complications were observed. CONCLUSION: Peripheral radial artery catheters are safe, are minimally invasive, and provide steady, high-flow rates, and they should be considered for patients requiring leukapheresis and lacking a suitable vascular access for drawing blood. PMID- 12559023 TI - Leukapheresis after high-dose chemotherapy and autologous peripheral blood progenitor cell transplantation: a novel approach to harvest a second autograft. AB - BACKGROUND: Autologous peripheral blood progenitor cells (PBPCs) are usually collected after the administration of conventional-dose chemotherapy (CDCT) and growth factors. However, there are no data available concerning the collection of PBPCs after high-dose chemotherapy (HDCT) and autologous hematopoietic transplantation in a larger series. STUDY DESIGN AND METHODS: Patients (n = 30) underwent leukapheresis for PBPC harvest after CDCT. After HDCT and autografting, the collection of a second PBPC autograft was attempted. RESULTS: Leukapheresis was performed after CDCT in all cases at a median of 118 CD34+ cells per microL (range, 18-589) and resulted in 6.4 x 10(6) CD34+ cells per kg (range, 1.7-29.0). After HDCT and autografting, 24 patients (80%) underwent secondary leukapheresis, although they had a significantly lower median of peripheral blood (PB) CD34+ cells (30/microL; range, 10-171; p < 0.001). In these patients a median of 3.6 x 10(6) CD34+ cells per kg (range, 1.6-10.1) was collected in the post transplantation course. In the remaining six patients (20%) with PB CD34+ cells < 10 per microL, no PBPC harvesting was performed. These so-called poor mobilizers had received significantly less CD34+ cells for autologous transplantation than patients with successful post-HDCT mobilization (median, 2.5 x 10(6)/kg [range, 1.7-3.0] vs. 6.5 x 10(6)/kg [range, 3.2-19.6]; p < 0.001). CONCLUSION: Collection of PBPCs is possible in most patients during the recovery phase of hematopoiesis after HDCT plus autografting, and the number of circulating PBPCs may be related to the CD34+ cell dose transfused by the preceding autograft. PMID- 12559025 TI - Where have all the donors gone? A personal reflection on the crisis in America's volunteer blood program. PMID- 12559024 TI - Trehalose ameliorates the cryopreservation of cord blood in a preclinical system and increases the recovery of CFUs, long-term culture-initiating cells, and nonobese diabetic-SCID repopulating cells. AB - BACKGROUND: The cryopreservation of HPCs in DMSO has been practiced by cord blood (CB) banks worldwide. Inevitably, some detriment to biologic function occurs as the result of freezing injuries and DMSO toxicity. Trehalose, a disaccharide, is a natural cryoprotectant in organisms capable of surviving extreme dehydration and cold. The objective of this study was to establish the cryopreservation of CB under preclinical conditions using trehalose as a supplement to DMSO. STUDY DESIGN AND METHODS: In a preclinical protocol, the effects of 5-percent trehalose with 10-percent DMSO or 5-percent DMSO on the cryopreservation of CB MNCs or nucleated cells (NCs) were further evaluated. The read-out system consisted of a panel of HPCs: early progenitors (CFU-GEMM, long-term culture-initiating cells [LTC-IC]) and committed progenitors (CFU-GM, CFU/BFU-E, CFU-megakaryocyte [CFU MK]). The homing and engraftment capacity of these cells were assessed in nonobese diabetic (NOD)-SCID mice. RESULTS: Trehalose increased the recoveries of CFU-GM, CFU/BFU-E, CFU-GEMM, and LTC-IC by over 7.25 percent (mean), 11.9 percent, 19.2 percent, and 12.9 percent, respectively, when compared with those in paired CB samples cryopreserved in 10-percent DMSO. Freezing and thawing reduced the yields of CFU-MK by 35.5 percent (mean) and 28.4 percent in MNC and NC samples, respectively, and the inclusion of 5-percent trehalose significantly retrieved these progenitor cells to over 90 percent of fresh samples. The improved recovery of functional HPLs was reflected by their multilineage engraftment in NOD-SCID mice. CONCLUSION: Trehalose at 5 percent significantly ameliorates the cryopreservation of CB progenitor cells at a preclinical protocol. The increased recoveries of these cells might potentially improve the engraftment outcomes of CB transplants. PMID- 12559026 TI - Hb-based oxygen carriers: are we there yet? PMID- 12559027 TI - HDN in a mother undergoing in vitro fertilization with donor ova. PMID- 12559028 TI - Jk(a-b-) and Kidd blood group genotypes in Chinese people. PMID- 12559029 TI - The causes and treatment of reactions to platelet transfusions. PMID- 12559032 TI - Organelle acidification and disease. AB - A subset of cellular compartments maintain acidic interior environments that are critical for the specific functions of each organelle and for cell growth and survival in general. The pH of each organelle reflects the balance between proton pumping, counterion conductance, and proton leak. Alterations in steady-state organelle pH due to defects in either proton pumping activity or counterion conductance have been suggested to contribute to the pathology of several diseases; however, definitive evidence remains elusive. This review describes recent evidence for the misregulation of organelle pH in the progression of cancer, Dent's disease, and cystic fibrosis. PMID- 12559033 TI - A kinetic proof-reading mechanism for protein sorting. AB - Resident proteins of the exocytic pathway are maintained at various levels through coatomer protein I (COPI)-mediated recycling. Sorting of cargo by COPI requires GTP hydrolysis by ADP-ribosylation factor 1 (ARF-1). This small GTPase recruits coatomer onto Golgi membranes and upon hydrolysis, is thought to release coatomer back into the cytosol. This step requires the activating protein, ARFGAP1. By coupling sorting to a cargo-induced sequestering of ARFGAP1, we have formulated a kinetic proof-reading model that explains how a GTP hydrolysis driven coat release can yield an active sorting event. The sorting scheme predicts a dependency on the amount of ARFGAP1 and explains the recent experimental findings that ARF-1 and COPI detach with different time constants from the Golgi membrane in vivo. PMID- 12559034 TI - Protein structure and import into the peroxisomal matrix. AB - Proteins destined for the peroxisomal matrix are synthesized in the cytosol, and imported post-translationally. It has been previously demonstrated that stably folded proteins are substrates for peroxisomal import. Mammalian peroxisomes do not contain endogenous chaperone molecules. Therefore, it is possible that proteins are required to fold into their stable, tertiary conformation in order to be imported into the peroxisome. These investigations were undertaken to determine whether proteins rendered incapable of folding were also substrates for import into peroxisomes. Reduction of albumin resulted in a less compact tertiary structure as measured by analytical centrifugation. Microinjection of unfolded albumin molecules bearing the PTS1 targeting signal resulted in their import into peroxisomes. Kinetic analysis indicated that native and unfolded molecules were imported into peroxisomes at comparable rates. While import was unaffected by treatment with cycloheximide, hsc70 molecules were observed to be imported along with the unfolded albumin molecules. These results indicate that proteins, which are incapable of assuming their native conformation, are substrates for peroxisomal import. When combined with previous observations demonstrating the import of stably folded proteins, these results support the model that tertiary structure has no effect on protein import into the peroxisomal matrix. PMID- 12559035 TI - Raft partitioning of the yeast uracil permease during trafficking along the endocytic pathway. AB - Lipid rafts, formed by the lateral association of sphingolipids and cholesterol in the external membrane leaflet, have been implicated in membrane traffic and cell signaling in mammalian cells. Yeast plasma membranes were also recently shown to contain lipid raft microdomains consisting of sphingolipids and ergosterol, and containing several plasma membrane proteins, including Gas1p, a GPI-anchored protein, and the [H+] ATPase Pma1p. In this study, we investigated whether lipid rafts were involved in the intracellular trafficking of a yeast transporter, uracil permease, which undergoes ubiquitin-dependent endocytosis. Regardless of its ubiquitination status, uracil permease was found to be associated with rafts in the plasma membrane. The expression of Fur4p in lcb1-100 cells, deficient in the first enzyme of sphingolipid synthesis, impaired the association of Fur4p with detergent-resistant fractions. When targeted to endocytic compartments, uracil permease appeared to be progressively transferred to detergent-soluble fractions, suggesting that the lipid environment might change between plasma membrane and endosomes. Consistent with this hypothesis, the wild-type form of the v-SNARE Snc1p, which is known to cycle between the plasma membrane and endosomal compartments, was recovered in both detergent resistant and detergent-soluble fractions. In contrast, a variant Snc1p that accumulates at the plasma membrane was recovered exclusively in detergent resistant fractions. PMID- 12559036 TI - CD2AP/CMS regulates endosome morphology and traffic to the degradative pathway through its interaction with Rab4 and c-Cbl. AB - The small GTPase Rab4 is involved in endocytosis through sorting and recycling early endosomes. To better understand the role of Rab4 in regulation of vesicular trafficking, we searched for effectors that specifically interact with Rab4-Q67L, the GTP-bound form of Rab4. We cloned an ubiquitous 80-kDa protein, identical to CD2-associated protein/Cas ligand with multiple SH3 domains (CD2AP/CMS), that interacts with Rab4-Q67L in the yeast two-hybrid system and in vitro. CD2AP/CMS expressed in mammalian cells was localized to punctate structures and along actin filaments. None of the known markers of early endosomes [Early Endosomes Antigen 1 (EEA1), Rab5 and Rab11] colocalized with the CD2AP/CMS-positive vesicles. However, coexpression of Rab4-Q67L with CD2AP/CMS induces a significant enlargement of EEA1-positive early endosomes. Rab4, CD2AP/CMS and Rab7 colocalized in these modified endosomes. Coexpression of c-Cbl and CD2AP/CMS also resulted in an enlargement of early endosomes. Using various truncated forms of CD2AP/CMS, we demonstrate that early endosomes enlargement requires that CD2AP/CMS interacts with both Rab4 and c-Cbl. The expression of a truncated form of CD2AP/CMS that retains the ability to interact with Rab4 but not c-Cbl inhibits ligand-induced PDGF receptor degradation. We propose that CD2AP/CMS, through interactions with Rab4 and c-Cbl, controls early endosome morphology and may play a role in traffic between early and late endosomes, and thus in the degradative pathway. PMID- 12559037 TI - Osteoclast ruffled border has distinct subdomains for secretion and degraded matrix uptake. AB - Subosteoclastic bone resorption is a result of HCl and proteinase secretion through a late endosome-like bone facing membrane domain called ruffled border. As bone matrix is degraded, it enters osteoclasts' transcytotic vesicles for further processing and is then finally exocytosed to the intercellular space. The present study clarifies the spatial relationship between these vesicle fusion and matrix uptake processes at the ruffled border. Our results show the presence of vacuolar H+-ATPase, small GTPase rab7 as well as dense aggregates of F-actin at the peripheral ruffled border, where basolaterally endocytosed transferrin and cathepsin K are delivered. On the contrary, rhodamine-labeled bone matrix enters transcytotic vesicles at the central ruffled border, where the vesicle budding proteins such as clathrin, AP-2 and dynamin II are also localized. We present a model for the mechanism of ruffled border turnover and suggest that, due to its late endosomal characteristics, the ruffled border serves as a valuable model for studying the dynamic organization of other endosomal compartments as well. PMID- 12559038 TI - The role of the ubiquitination-proteasome pathway in breast cancer: applying drugs that affect the ubiquitin-proteasome pathway to the therapy of breast cancer. AB - The ubiquitin-proteasome pathway is responsible for most eukaryotic intracellular protein degradation. This pathway has been validated as a target for antineoplastic therapy using both in vitro and preclinical models of human malignancies, and is influenced as part of the mechanism of action of certain chemotherapeutic agents. Drugs whose primary action involves modulation of ubiquitin-proteasome activity, most notably the proteasome inhibitor PS-341, are currently being evaluated in clinical trials, and have already been found to have significant antitumor efficacy. On the basis of the known mechanisms by which these agents work, and the available clinical data, they would seem to be well suited for the treatment of breast neoplasms. Such drugs, alone and especially in combination with current chemotherapeutics, may well represent important advances in the therapy of patients with breast cancer. PMID- 12559040 TI - The role of the ubiquitination-proteasome pathway in breast cancer: use of mouse models for analyzing ubiquitination processes. AB - Turnover of several regulatory proteins results from targeted destruction via ubiquitination and subsequent degradation through the proteosome. The timely and irreversible degradation of critical regulators is essential for normal cellular function. The precise biochemical mechanisms that are involved in protein turnover by ubiquitin-mediated degradation have been elucidated using in vitro assays and cell culture systems. However, pathways that lead to ubiquitination of critical regulatory proteins in vivo are more complex, and have both temporal and tissue-specific differences. In vivo models will allow identification of substrates and enzymes of the ubiquitin-proteosome pathway that play important roles in selected tissues and diseases. In addition, assessment of the therapeutic efficacy of drugs designed to inhibit or enhance protein turnover by ubiquitination requires in vivo models. In the present review we describe selected examples of transgenic and knockout models of proteins that are known either to be regulated by ubiquitin-mediated degradation or to have a catalytic function in this process, and to play an important role in breast cancer. We outline the functions of these proteins in vivo and focus on knowledge gained in the comparison of in vivo behavior predicted from cell-free in vitro data or from experiments conducted in cell culture systems. PMID- 12559039 TI - The role of the ubiquitination-proteasome pathway in breast cancer: ubiquitin mediated degradation of growth factor receptors in the pathogenesis and treatment of cancer. AB - Aberrant activity of growth factor receptors has been implicated in the pathogenesis of a wide variety of malignancies. The negative regulation of signaling by growth factor receptors is mediated in large part by the ubiquitination, internalization, and degradation of the activated receptor. Over the past few years, considerable insight into the mechanisms that control receptor downregulation has been gained. There are also data suggesting that mutations that lead to inhibition of downregulation of growth factor receptors could play a role in the pathogenesis of cancer. Therapies directed at enhancing the degradation of growth factor receptors offer a promising approach to the treatment of malignancies. PMID- 12559041 TI - Expression profiling to predict outcome in breast cancer: the influence of sample selection. AB - Gene expression profiling of tumors using DNA microarrays is a promising method for predicting prognosis and treatment response in cancer patients. It was recently reported that expression profiles of sporadic breast cancers could be used to predict disease recurrence better than currently available clinical and histopathological prognostic factors. Having observed an overlap in those data between the genes that predict outcome and those that predict estrogen receptor alpha status, we examined their predictive power in an independent data set. We conclude that it may be important to define prognostic expression profiles separately for estrogen receptor-alpha-positive and estrogen receptor-alpha negative tumors. PMID- 12559042 TI - BAD: a good therapeutic target? AB - The major goal in cancer treatment is the eradication of tumor cells. Under stress conditions, normal cells undergo apoptosis; this property is fortunately conserved in some tumor cells, leading to their death as a result of chemotherapeutic and/or radiation-induced stress. Many malignant cells, however, have developed ways to subvert apoptosis, a characteristic that constitutes a major clinical problem. Gilmore et al. recently described the ability of ZD1839, a small-molecule inhibitor of the epidermal growth factor receptor (EGFR), to induce apoptosis of mammary cells that are dependent upon growth factors for survival. Furthermore, they showed that the major effector of the EGFR-targeted therapy is BAD, a widely expressed BCL-2 family member. These results are promising in light of the role of the EGFR in breast cancer development. PMID- 12559043 TI - Host microenvironment in breast cancer development: inflammatory cells, cytokines and chemokines in breast cancer progression: reciprocal tumor-microenvironment interactions. AB - A comprehensive overview of breast cancer development and progression suggests that the process is influenced by intrinsic properties of the tumor cells, as well as by microenvironmental factors. Indeed, in breast carcinoma, an intensive interplay exists between the tumor cells on one hand, and inflammatory cells/cytokines/chemokines on the other. The purpose of the present review is to outline the reciprocal interactions that exist between these different elements, and to shed light on their potential involvement in breast cancer development and progression. PMID- 12559044 TI - Complex roles for telomeres and telomerase in breast carcinogenesis. AB - Telomerase - an enzyme that endows cells with unlimited proliferative potential - is differentially expressed in cancer cells and in normal cells. Although most primary human cells lack telomerase, the enzyme is upregulated in more than 90% of invasive breast cancers. As a result, much of breast cancer development occurs before telomerase is reactivated during a critical transition from a telomerase negative to a telomerase-positive state. During this transition, the telomere shortening that accompanies cell division may either prevent or facilitate tumorigenesis by activating checkpoints and impairing chromosomal stability. In mature cancers, telomerase probably serves a crucial role in tumor progression and maintenance by stabilizing telomeres and supporting the immortal growth of breast cancer cells. PMID- 12559045 TI - Intrauterine environment, mammary gland mass and breast cancer risk. AB - Two intimately linked hypotheses on breast cancer etiology are described. The main postulate of the first hypothesis is that higher levels of pregnancy estrogens and other hormones favor the generation of a higher number of susceptible stem cells with compromised genomic stability. The second hypothesis postulates that the mammary gland mass, as a correlate of the number of cells susceptible to transformation, is an important determinant of breast cancer risk. A simple integrated etiological model for breast cancer is presented and it is indicated that the model accommodates most epidemiological aspects of breast cancer occurrence and natural history. PMID- 12559047 TI - Department of Defense Era of Hope meeting, Orlando, Florida, USA, 25-28 September 2002. AB - The Era of Hope meeting addressed with a multidisciplinary approach the most critical issues in breast carcinogenesis. The issues that we summarize here include: a) the use of rodent models for the study of mammary gland development and breast tumorigenesis; b) the effects of stroma on mammary epithelial differentiation and malignant transformation; c) a further characterization of the interactions between steroid and growth factor receptors; d) the improvement of technologies for early detection of breast tumors and the establishment of their progression; and e) the development of vaccines as potential new therapies against specific tumor markers. PMID- 12559046 TI - Regulation of BRCA1 expression and its relationship to sporadic breast cancer. AB - Germ-line mutations in the BRCA1 tumour suppressor gene contribute to familial breast tumour formation, but there is no evidence for direct mutation of the BRCA1 gene in the sporadic form of the disease. In contrast, decreased expression of the BRCA1 gene has been shown to be common in sporadic tumours, and the magnitude of the decrease correlates with disease progression. BRCA1 expression is also tightly regulated during normal breast development. Determining how these developmental regulators of BRCA1 expression are co-opted during breast tumourigenesis could lead to a better understanding of sporadic breast cancer aetiology and the generation of novel therapeutic strategies aimed at preventing sporadic breast tumour progression. PMID- 12559048 TI - Expression profiling predicts outcome in breast cancer. PMID- 12559049 TI - Centrosome cycle studies reveal promising candidates for anti-cancer drug design. PMID- 12559051 TI - Functional and molecular characterisation of mammary side population cells. AB - BACKGROUND: Breast cancer is thought to arise in mammary epithelial stem cells. However, the identity of these stem cells is unknown. METHODS: Studies in the haematopoetic and muscle systems show that stem cells have the ability to efflux the dye Hoechst 33342. Cells with this phenotype are referred to as the side population (SP). We have adapted the techniques from the haematopoetic and muscle systems to look for a mammary epithelial SP. RESULTS: Of mammary epithelial cells isolated from both the human and mouse mammary epithelia, 0.2-0.45% formed a distinct SP. The SP was relatively undifferentiated but grew as typical differentiated epithelial clones when cultured. Transplantation of murine SP cells at limiting dilution into cleared mammary fat pads generated epithelial ductal and lobuloalveolar structures. CONCLUSION: These data demonstrate the existence of an undifferentiated SP in human and murine mammary epithelium. Purified SP cells are a live single-cell population that retain the ability to differentiate in vitro and in vivo. Studies of haematopoetic cells have suggested that the SP phenotype constitutes a universal stem cell marker. This work therefore has implications for mammary stem cell biology. PMID- 12559052 TI - Expression of glucocorticoid and progesterone nuclear receptor genes in archival breast cancer tissue. AB - BACKGROUND: Previous studies in our laboratory have shown associations of specific nuclear receptor gene variants with sporadic breast cancer. In order to investigate these findings further, we conducted the present study to determine whether expression levels of the progesterone and glucocorticoid nuclear receptor genes vary in different breast cancer grades. METHODS: RNA was extracted from paraffin-embedded archival breast tumour tissue and converted into cDNA. Sample cDNA underwent PCR using labelled primers to enable quantitation of mRNA expression. Expression data were normalized against the 18S ribosomal gene multiplex and analyzed using analysis of variance. RESULTS: Analysis of variance indicated a variable level of expression of both genes with regard to breast cancer grade (P = 0.00033 for glucocorticoid receptor and P = 0.023 for progesterone receptor). CONCLUSION: Statistical analysis indicated that expression of the progesterone nuclear receptor is elevated in late grade breast cancer tissue. PMID- 12559053 TI - Lack of evidence for an association of Epstein-Barr virus infection with breast carcinoma. AB - BACKGROUND: Epstein-Barr virus (EBV) is a ubiquitous human gamma-herpes virus infecting more than 90% of the population worldwide. EBV is associated with certain malignancies (e.g. Burkitt lymphoma, Hodgkin lymphoma and nasopharyngeal carcinoma). Recent studies have raised the possibility that EBV may also be involved in the pathogenesis of breast carcinoma, the most common carcinoma of females. If substantiated, this finding would have major implications regarding prevention and therapy of the disease. The studies published so far have employed diverse methods, however, and the results have been controversial. METHODS: Using the EBV DNA PCR, EBV DNA in situ hybridisation and in situ hybridisation for the detection of the EBV-encoded RNAs, and using immunohistochemistry for the demonstration of the EBV-encoded nuclear antigen 1, we have studied a series of 59 invasive breast carcinomas for evidence of EBV infection. RESULTS: EBV-encoded RNA-specific in situ hybridisation and EBV-encoded nuclear antigen 1 immunohistochemistry were negative in all cases. Using the PCR, EBV DNA was detected in four out of 59 cases. These cases were further studied by EBV DNA in situ hybridisation, showing an absence of viral DNA from the tumour cells. CONCLUSION: These results indicate that breast carcinoma is not an EBV-associated tumour. PMID- 12559054 TI - Gingival prostheses--a review. AB - Gingival replacement is often a component of comprehensive prosthodontics. Gingival prostheses may be fixed or removable and may be made from acrylics, composite resins, silicones or porcelain-based materials. Undercuts or dental attachments are used to secure removable prostheses, which are esthetically pleasing and easy to maintain. This paper describes several clinical situations in which gingival prostheses were used effectively. PMID- 12559055 TI - A modified impression technique for accurate registration of peri-implant soft tissues. AB - Replacement of single missing teeth with an implant-supported restoration is recognized as a highly successful treatment. An impression technique for peri implant soft-tissue replication in an anterior zone is described. The technique involves use of an interim restoration as an abutment for the final impression. This allows accurate duplication of the soft tissues and fabrication of a final restoration with the correct emergence profile. PMID- 12559056 TI - Dental erosion in gastroesophageal reflux disease. AB - Dentists are often the first health care professionals to diagnose dental erosion in patients with gastroesophageal reflux disease (GERD). Gastroesophageal reflux (GER) is the passage of gastric contents into the esophagus, and GERD is defined as symptoms or complications of GER. Twenty-four-hour monitoring of esophageal pH is helpful in diagnosing GERD. Treatment of dental erosion resulting from GERD involves a multidisciplinary approach among family physician, dentist, prosthodontist, orthodontist and gastroenterologist. When possible, dental erosion should be treated with minimal intervention, and such treatment should include control of microflora, remineralization, adhesive restorations and use of biomimetic materials. PMID- 12559057 TI - Discuss before fabricating: communicating the realities of partial denture therapy. Part I: patient expectations. AB - During planning for treatment involving cast removable partial dentures (RPDs), it is important to remember that the 2 major reasons why patients seek this treatment are to improve esthetics and to improve mastication. Treatment that is considered unsuccessful may not necessarily represent poor execution of procedures, but rather may not have fulfilled the patient's expectations with regard to these 2 goals. Searches of the Cochrane Collaboration and MEDLINE databases were conducted to identify issues pertaining to patient compliance in wearing cast RPDs. Compliance was better when the patient's chief concern was replacement of anterior teeth. The success of RPDs for replacement of teeth missing from other areas depended on the patient's perception of the value of such replacement. Maximum masticatory efficiency is equivalent for people functioning from second premolar to second premolar and those wearing partial dentures. Many clinicians believe that the prescription of partial dentures which replace teeth missing distally to the second premolars represents overtreatment. PMID- 12559058 TI - Discuss before fabricating: communicating the realities of partial denture therapy. Part II: clinical outcomes. AB - The premise of this review is that patients' satisfaction (and hence compliance) with partial denture therapy may be better if they are more fully informed about the limitations of the prosthesis they are to receive. Neither the dentist nor the patient should assume that all of their respective expectations will be mutually understood and inherently met. By discussing patient-centred issues and predictable clinical outcomes, both dentist and patient will be better prepared to determine whether a removable prosthesis is appropriate. Searches of the Cochrane Collaboration and MEDLINE databases were conducted to identify issues pertaining to patient compliance in wearing cast removable partial dentures. In addition to the 2 most frequent patient concerns, esthetics and mastication, discussed in the first article of this series, additional aspects of concern to the dentist and the patient when considering a removable partial denture include overeruption, post-insertion care, comfort, longevity of the prosthesis, effect on speech and biologic consequences are discussed here. PMID- 12559059 TI - Perspectives on intake of calcium-rich foods among Asian, Hispanic, and white preadolescent and adolescent females. AB - OBJECTIVE: Declining calcium intake among adolescents warrants attention. Our objective was to identify influences on adolescents' consumption of calcium-rich foods. DESIGN: Focus groups were conducted with girls representing 2 age groups (11 to 12 or 16 to 17 years) and 3 macroethnic groups (Asian, Hispanic, or white). SETTING: Public schools in 10 states. PARTICIPANTS: A convenience sample (n = 200) was recruited through schools. VARIABLES MEASURED: Focus groups (n = 35) were audiotaped and transcribed. Influences relative to consumption of milk or other calcium-rich foods were identified. ANALYSIS: Comments were coded as motivators or barriers within each focus group. Content analysis procedures were used to compare ethnic and age groups. RESULTS: A barrier to milk consumption that was more common among older girls and Asian groups was the limited expectation within families for drinking milk. Many controlled their own beverage choices, and milk, even if liked, was only one option. Milk was positively associated with strength and bone health, but these attributes were viewed as being more important for boys than girls. Milk was associated with breakfast, school lunches, cereal, and desserts. White girls had the most positive reactions to milk and Hispanic girls the most negative. All groups were positive toward pizza, ice cream, and cheese. CONCLUSIONS AND IMPLICATIONS: To improve calcium intake among teens, interventions should include a family component, stress the benefits of milk for girls, and focus on breakfast. PMID- 12559060 TI - Using computer-based assessments to evaluate interactive multimedia nutrition education among low-income predominantly Hispanic participants. AB - OBJECTIVE: This research was conducted to measure the effectiveness of interactive multimedia (IMM) with low-income Hispanic persons. DESIGN: The effectiveness of the program was examined using a quasi-experimental pretest/post test control group design. SETTING: Subjects were recruited from nutrition, health, and English as a Second Language (ESL) programs in Colorado. PARTICIPANTS: Thirty-six intervention and 34 control participants formed a convenience sample of low-income and predominantly Hispanic persons. INTERVENTION: Intervention participants received a 15-minute module about breakfast. Control participants received a non- nutrition-related IMM module. MAIN OUTCOME MEASURES: Primary variables included knowledge, attitude, and stage of change scores. ANALYSIS: Paired and independent sample t tests, chi-square analysis, and repeated-measures analysis of covariance (ANCOVA) were used. RESULTS: Intervention participants significantly increased knowledge, attitude, and total scores (P <.001) between pretest and post-test and had significantly greater increases than the control group (P <.001). CONCLUSIONS: The results support using IMM to disseminate nutrition education to the target population and the feasibility of using computer-based questionnaires to evaluate the effectiveness of IMM nutrition education programs. IMPLICATIONS: This research provides the basis for the continued development of computer-based assessment tools. PMID- 12559061 TI - Development and validation of a shelf inventory to evaluate household food purchases among older adults with diabetes mellitus. AB - OBJECTIVE: Valid measures of behavioral outcomes are needed to evaluate interventions. The purpose of this research was to validate a shelf inventory and evaluate food purchases using the inventory among older adults with diabetes. DESIGN: A 166-item shelf inventory of household food was assessed for face, content, and criterion validity. The sensitivity and specificity of the inventory were determined by comparing participant- and interviewer-completed inventories. A randomized pretest/post-test control group design was used to evaluate household food purchases following an intervention. SETTING: The intervention was held in an outpatient setting. PARTICIPANTS: Study participants were > or = 65 years old with type 2 diabetes mellitus for > or = 1 year. INTERVENTION: The intervention included 10 weekly group sessions and emphasized applying food label information to food purchases. MAIN OUTCOME MEASURES: Each food on the inventory was defined as "encouraged" or "discouraged" according to intervention messages. ANALYSIS: Inventory sensitivity was the proportion of foods present in households that were accurately identified on the self-reported inventory as present; specificity was the proportion of foods not present that were accurately identified as not present. Two-sample t tests compared intervention scores for encouraged/discouraged foods by treatment group. RESULTS: Overall sensitivity and specificity were.90 and.97, respectively. Intervention results found more encouraged than discouraged foods present at post-test (t [91] = 2.3, P =.02). CONCLUSIONS AND IMPLICATIONS: The shelf inventory is a sensitive, specific, and valid tool for assessing household food purchases and can be used to evaluate food choice interventions among older adults with type 2 diabetes mellitus. PMID- 12559062 TI - Usefulness of the food habits questionnaire in a worksite setting. AB - OBJECTIVE: To examine the validity and reliability of the 24-item Food Habits Questionnaire (FHQ) in a worksite setting. DESIGN: In a longitudinal design, subjects in a 9-month worksite intervention program completed the FHQ pre- and postintervention. A randomly selected subsample also completed 2 sets of 4-day food records. SETTING: Three locations within a multisite industrial equipment company. PARTICIPANTS: Subjects (N = 178) self-selected to participate in the program and completed the FHQ. Mean age was 40.7+/-10.6, 60% were male, and 82% were white, and there were 15 occupational categories. Thirty-two subjects completed 2 sets of 4-day food records. MAIN OUTCOME MEASURES: To examine the validity and reliability of the FHQ total scale and subscales in a worksite setting with both men and women from varying socioeconomic levels. ANALYSIS: Cronbach a coefficient to estimate internal consistency for the total scale and the subscales, Pearson correlation coefficients to estimate test-retest reliabilities and criterion validity. RESULTS: For the 5 subscales, findings showed generally low internal consistency, moderate test-retest reliability, and low to moderate correlations with food record results. Psychometric properties for the total score of the FHQ included satisfactory internal consistency, test retest reliability, and criterion validity. CONCLUSIONS AND IMPLICATIONS: The FHQ subscales did not perform well and need to be more fully developed. Therefore, the FHQ may best be used as a total scale score to indicate fat-related behaviors. PMID- 12559063 TI - Economic analysis of bilingual interactive multimedia nutrition education. AB - CONTEXT: Interactive multimedia (IMM) provides a means to deliver accurate, consistent, and convenient nutrition education. OBJECTIVE: To compare direct costs of bilingual IMM and peer nutrition education methods. DESIGN: Cost per unit of nutrition education for each delivery method was calculated. SETTING: Touch-screen computer kiosk systems in food assistance programs. PARTICIPANTS: Approximately 727 low-income and Hispanic persons. INTERVENTION: Bilingual IMM and nutrition education classes taught by peer educators. MAIN OUTCOME MEASURES: Cost per unit of nutrition education delivered. ANALYSIS: Descriptive: to compare costs and determine break-even points. RESULTS: Initially, costs for both methods dropped and then nearly leveled for the peer method at a break-even point of 633 units, whereas those for IMM continued to decrease. CONCLUSIONS: Interactive multimedia is a cost-effective method of delivering nutrition education, especially to large numbers of people. IMPLICATIONS: Use of this technology could reach additional low-income persons at decreasing costs. PMID- 12559064 TI - Snack consumption and waste by preschool children served "cute" versus regular snacks. AB - OBJECTIVE: To determine if preschool children would eat and waste different amounts of food when served snacks in a regular versus a "cute" form. DESIGN: Preschool children were served 6 snacks in a cute form and the same snacks in a regular form using the same food components during 24 snack sessions. Children served as their own controls. SUBJECTS/SETTING: Thirty-nine children ages 36 to 60 months (20 girls, 19 boys) in a university child development laboratory. MAIN OUTCOME MEASURES: Snack portions eaten and wasted and time for eating. ANALYSIS: Data were analyzed using a 2 3 6 factorial analysis of variance, with snack form (cute or regular) and snack components as the 2 factors. RESULTS: There was no significant difference in children's snack consumption or waste between cute and regular versions of snacks. It took children an average of 2 minutes longer to eat the cute snacks. CONCLUSIONS AND IMPLICATIONS: There is no apparent advantage to serving cute snacks in terms of increased consumption or decreased waste. Further research should be conducted on children's mealtime behaviors when served cute foods to learn more about children's responses to the overall appearance of their food. PMID- 12559065 TI - Weight control brochures for five target audiences of women ages 25 to 40. PMID- 12559068 TI - Intramammary sentinel nodes in early breast cancer: can we find them and do they matter? AB - Intramammary nodes identified using the sentinel node biopsy technique can play an important prognostic role in early breast cancer. Two cases of intramammary nodes found by sentinel node biopsy are discussed from the Guildford perspective. PMID- 12559067 TI - A common voice for cancer surgery. PMID- 12559069 TI - Radiofrequency ablation in the management of liver tumours. PMID- 12559070 TI - Does local surgery have a role in the management of stage IV breast cancer? AB - AIMS: There are no uniformly agreed guidelines regarding the treatment of local breast cancer in patients who have stable metastatic disease. The aim of this study was to define the role of breast surgery in the management of stage IV disease by reviewing the clinical outcome in patients with stage IV disease submitted to surgery in a regional breast cancer unit. METHODS: All patients who underwent breast surgery from 1993 to 1999 and had known metastatic disease or who were diagnosed with metastases within one month of surgery were identified and their clinical outcome was studied using death and local recurrence as end points. RESULTS: Median survival after breast surgery was 23 months. Ten of the 20 patients were alive with no local disease at 20 months mean follow-up. Three of 10 patients who died developed local recurrence and had local disease at the time of death. CONCLUSION: The local surgery does have a role in controlling the primary cancer and controlling local symptoms in a selected group of patients with stable metastatic disease. PMID- 12559071 TI - Objective measurement of breast cancer oestrogen receptor status through digital image analysis. AB - AIMS: The authors have previously described quantitative, computer-assisted analysis of oestrogen receptor status in immuno-histochemically stained sections in patients with primary breast cancer. The aim of this study was to validate the aforementioned system against the commonly used methods of assessing oestrogen receptor status. METHODS: Paraffin embedded sections from 156 patients with primary breast carcinoma were stained with anti-alpha-oestrogen receptor monoclonal antibody (1D5) using a standard immunohistochemical protocol. Images from 10 high-powered fields were captured from each section using a digital camera mounted on a microscope and analyzed using Adobe Photoshop image analysis software. A nuclear mask was obtained by digitally selecting the nuclear area. Staining intensity in the nuclear mask was then analyzed using red-scale absorption characteristics. Manual assessment of oestrogen receptor status was performed through counting the percentages of cells that are positive from 200 randomly sampled nuclei from ten high powered fields HPF. Cut off value for positivity was taken as 10%. Cytosolic oestrogen receptor concentration was measured through enzyme immunisation. Cut off value for ER positivity was taken as 200 fmol/g (wet tissue). RESULTS: One hundred and fifty-six sections were studied of which 41 were ER negative. Median percentage positivity in the remainder was 90% (17-100) by manual assessment. The median red scale value was 108 (58-156). A close correlation was observed between median optical density of the nuclear mask and percentage positivity assessed manually (P<0.0001). There was a significant correlation between the optical density of the nuclear mask and cytosolic oestrogen receptor concentration (P<0.001). CONCLUSION: Oestrogen receptor positivity can be accurately assessed through digital image analysis. This process offers objective data regarding the amount of oestrogen receptors within the nuclei as well as the percentage of nuclei, which express oestrogen receptors. PMID- 12559072 TI - Intraoperative evaluation of axillary sentinel lymph nodes using touch imprint cytology and immunohistochemistry: I. Protocol of rapid immunostaining of touch imprints. AB - AIMS: Sentinel Node Biopsy (SNB) is considered an accurate method of detecting axillary lymph node status in patients with small breast cancer. Combined with an accurate and rapid histopathology tool, it could spare this group of patients unnecessary Axillary Node Clearance (ANC) with its associated hazards. Intraoperative examination of SNB for cancer cells has been investigated using both Frozen Sections (FS) and Imprint Cytology (IC) stained with different stains. This study is devoted to establish a reliable and rapid protocol for immunostaining of touch imprints from SNB. METHODS: We investigated two different EPOS (Enhanced Polymer One-Step staining--DAKO) anticytokeratin antibodies, five different tissue fixatives and different incubation periods and temperatures with both positive and negative controls. RESULTS: We have developed a protocol, which produced good and consistent immunostaining of touch imprints. The initial results using this protocol are concordant with those of permanent Haematoxylin and Eosin (H&E) sections. CONCLUSIONS: We propose this protocol for rapid immunostaining of touch imprints of SNB. PMID- 12559073 TI - Mitotic activity index in interval breast cancers. AB - AIMS: The Mitotic Activity Index (MAI) is a strong prognostic factor for disease free survival in breast cancer. The MAI is lower in screen detected tumours, correlating with less aggressive biological behaviour in this group. In this study the MAI is compared between screen detected, interval and symptomatic breast cancers. METHODS: Between 1991 and 1999, the MAI was determined in 581 breast cancers, 160 were detected by screening, 66 were interval carcinomas, and 355 were symptomatic breast cancers. Other prognostic factors were also registered. RESULTS: The interval group had a significantly higher median MAI (17 18, range 1-134) than the screen detected group (7-8, range 0-94,P <0.0001). There was no difference with the symptomatic group (MAI 15, range 0-149,P =0.92). CONCLUSIONS: Interval cancers had an intermediate outcome when correlated with other prognostic factors, compared to screen detected and symptomatic cancers. PMID- 12559074 TI - Dynamic contrast enhanced magnetic resonance imaging aids the surgical management of invasive lobular breast cancer. AB - AIMS: Infiltrating Lobular Carcinoma (ILC) accounts for 5-10% of all breast cancers and is associated with subtle clinical and mammographic changes. It is also frequently multifocal and traditional diagnostic methods are unable to reliably detect this preoperatively. The aim of the study was to evaluate the efficacy of current imaging modalities compared with dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) in the evaluation of ILC. METHODS: Data from women with ILC treated between 1996 and 2000 who had undergone preoperative triple assessment and DCE-MRI was analysed. The diagnostic accuracy and the detection of multifocal disease for each modality was assessed and any changes in clinical management following DCE-MRI were recorded. RESULTS: Twenty-one women (mean age 57, range 43-72 years) were recruited. The sensitivity of X-ray mammography, ultrasound, clinical assessment, fine needle aspiration cytology or core biopsy and DCE-MRI was 90.5, 87.5, 76.2, 85.7 and 95.2% respectively. DCE MRI identified all the patients with subsequently histologically proven multifocal disease (n=11) with PPV and NPV of 100 and 95.2% respectively. X-ray mammography and ultrasound combined identified only 27% (n=3/11) with PPV and NPV of 100 and 55.6% respectively. Management was changed in 24% (5/21) of the cases following DCE-MRI. CONCLUSION: DCE-MRI has a higher sensitivity than other imaging modalities and is able to accurately delineate multifocal disease not evident on conventional imaging. DCE-MRI should therefore be used for accurate staging prior to surgery for invasive lobular breast cancer. PMID- 12559075 TI - Detection of genetic abnormalities in neoplasms from Greek patients with FAP. AB - AIMS: DNA microsatellite instability is a well-known feature of hereditary non polyposis colon cancer; however, its incidence in familial adenomatous polyposis, is unclear. We report the frequency of microsatellite instability and other genetic abnormalities in a group of Greek patients with FAP, in relation to various clinicopathological variables. METHODS: Thirty-four tissue specimens from 10 patients with FAP were studied. Microsatellite instability was investigated at six loci: BAT25, BAT26, D2S123, D5S346, D17S250 and TGF-beta RII poly(A) tract. p53 and K-ras mutations were also examined. RESULTS: Microsatellite instability was detected in two FAP adenocarcinomas from different patients. Mutation percentages observed were: in K-ras 45% and 50% and in p53 14% and 58%, of FAP adenomas and adenocarcinomas, respectively. No K-ras or p53 mutations were determined in the two microsatellite instable adenocarcinomas. CONCLUSION: Microsatellite instability is detectable in a small proportion of adenocarcinomas complicating FAP. This minority of cases may constitute a distinct subgroup among FAP neoplasms. PMID- 12559076 TI - Prognostic significance of caspase-3 expression in primary resected esophageal squamous cell carcinoma. AB - AIMS: The present study retrospectively examines the expression of caspase-3 in primary resected esophageal squamous cell carcinoma (ESCC) and the correlation between the outcome of patients and the expression of proteins. METHODS: Immunohistochemistry and Western blot analyses were used to analyse the expression of caspase-3 in 40 archival specimens of patients with primary resected ESCC. RESULTS: According to our cut-off point of the staining for caspase-3, 24 (60%) cases were positive and 16 (40%) negative. Caspase-3 expression correlated with a significant favorable prognosis in primary resected ESCC (P=0.02). A multivariate analysis of clinical and biological factors indicated that stage, tumor differentiation, and caspase-3 expression were independent prognostic factors. CONCLUSIONS: Caspase-3 expression might be a good and new prognostic indicator for primary resected ESCC. PMID- 12559077 TI - The effect of chemotherapy on the morphology of the growth plate and metaphysis of the growing skeleton. AB - AIM: To establish the effect of three single chemotherapeutic agents on the growing skeleton, male Wistar rats were studied. METHODS: From the age of 4 weeks the rats were given iv doxorubicin (DOX) 15 mg/m(2) body surface area (BSA), methotrexate (MTX) 60 mg/m(2) BSA or cisplatin (CDDP) 7.5 mg/m(2) BSA. One non treated control group was fed ad libitum (ad lib) and for every drug-treated group there was a diet-control group. After dissection at 13 weeks of age, morphology of the proximal tibial growth plate and metaphysis were studied. RESULTS: Compared to the ad lib group, DOX significantly decreased and MTX increased growth plate height (P<0.05). CDDP decreased height of the proliferating layer (P<0.05). Trabecular volume was decreased in the DOX and CDDP treated rats compared to the ad lib group (P=0.054). Compared to the diet control group trabecular bone volume was unaffected in the DOX group and decreased in the MTX and CDDP group (P<0.05). CONCLUSIONS: Doxorubicin causes growth plate thinning, methotrexate increases growth plate height and cisplatin does not affect growth plate height. All three chemotherapeutic agents decrease the trabecular volume of the proximal tibial metaphysis. Part of the effect of DOX, MTX and CDDP is related to the treatment induced malnutrition. PMID- 12559078 TI - The swing procedure for pelvic ring reconstruction following tumour excision. AB - AIM: High grade periacetabular osteosarcomas extending to the sacro-iliac region present a difficult management problem. We describe our experience of the swing procedure as a method of limb salvage. METHOD: The procedure was performed on three patients with stage II B osteosarcoma. RESULTS: Functional outcome was poor in all three with a mean Musculoskeletal Tumor Society score of 29% (27-30) at last review. All three developed local recurrence and one developed pulmonary metastases. The mean disease free interval post-operatively was 4.7 months (4-6) and they died of their disease at a mean of 10 months (5-16). CONCLUSIONS: This form of limb salvage is technically demanding and carries a high morbidity. Functional outcome is poor and the prognosis for this group of patients is not improved by surgery. Alternative treatment options are discussed. PMID- 12559079 TI - Use of the Ilizarov technique to improve limb function following hemipelvectomy. AB - AIMS: Limb salvage surgery combined with tumour resection has become established in the management of pelvic tumours. However, subsequent reconstructive options for maintenance of quality of life lead to varied outcomes. We present a hitherto undescribed use of the Ilizarov technique as a second stage adjunct, for the optimisation of function, after first stage tumour resection and arthrodesis. METHODS: We describe the surgical technique used to address leg length discrepancy and abductor dysfunction following internal hemipelvectomy with ischifemoral arthrodesis. Distal femoral and distal tibial lengthening using circular frames, with a valgus subtrochanteric femoral osteotomy, enabled a rapid correction of both anomalies. Controlled varus correction at the distal femoral osteotomy site was performed to obtain a horizontal joint line at the knee. CONCLUSION: The Ilizarov technique is suitable for selected patients with tumour free survival following the initial resection. Our experience indicates that it is a good alternative to a mammoth one stage internal hemipelvectomy with reconstruction. The latter is often attempted even though there is a high risk of local recurrence and distant metastasis. PMID- 12559080 TI - Peritoneal cancer index: a prognostic indicator of survival in advanced ovarian cancer. AB - AIM: The peritoneal cancer index (PCI) has been used for the detailed evaluation of the peritoneal spread in tumors of gastrointestinal origin and has been found to be a prognostic indicator of survival. The aim of this study was the identification of the significance of the peritoneal cancer index in advanced ovarian cancer. METHODS: From 1990 to 2001, 60 women, mean age 65+/-10.84 (41 86), were treated for advanced ovarian cancer. The performance status (Karnofsky performance scale), age, prior surgery score (PSS), peritoneal cancer index (PCI), tumor volume, tumor grade, residual tumor, the presence of ascites, treatment with adjuvant chemotherapy, histopathologic subtype and FIGO stage were retrospectively correlated to survival using univariate model of statistical analysis. RESULTS: Hospital mortality and morbidity were 11.7 and 16.7% respectively. The recurrence rate was 23.3%. Overall 5-year survival rate was 41% and mean survival 63+/-8 months. The peritoneal cancer index was related to survival (P=0.0253). The other favorable clinical prognostic indicators of survival were low grade and small volume tumors, treatment with adjuvant chemotherapy and complete cytoreductive surgery (P<0.05). CONCLUSIONS: The peritoneal spread in advanced ovarian cancer can be assessed in detail using the peritoneal cancer index. It is a significant prognostic factor of survival and is useful in identifying subgroups. PMID- 12559081 TI - Expression of cell-cycle regulatory proteins and their prognostic value in superficial low-grade urothelial cell carcinoma of the bladder. AB - AIMS: Cell-cycle regulatory proteins are important indicators in determining progression trough the cell-cycle and progression to invasive cancer in patients presenting with superficial bladder cancer. We performed an immunohistochemical study in order to evaluate the prognostic value of the expression of p16, p27, pRb, p53 and Ki-67 in superficial grade I and II papillary urothelial cell carcinoma of the bladder. METHODS: p16, p27, p53, pRb and Ki-67 immunoexpression was studied in 14 pTa, 35 pT1a and 7 pT1b bladder tumours at presentation and at recurrence of their tumours. The recurrence-free survival and the progression free survival were analysed according to these regulatory cell-cycle proteins expression. RESULTS: For survival in univariate analysis a high Ki-67 labelling index was a poor prognostic factor for recurrence-free and progression-free survival (P=0.0014 and P=0.012, respectively). Ki-67 labelling index was also an independent recurrence-free survival prognostic factor (P=0.0005). The p16, p27, p53 and pRb immunoreactivity was not significantly associated with recurrence or progression rate in this group of bladder carcinomas. CONCLUSIONS: These data suggest that the Ki-67 labelling index can be a reliable marker in predicting recurrence and/or progression in superficial low-grade bladder carcinomas and may be relevant in planning adjuvant therapy. PMID- 12559082 TI - Treatment results of regional metastasis from cutaneous head and neck squamous cell carcinoma. AB - AIMS: To investigate the results of our treatment policy, we present our institutional experience in the management of regional neck node metastases of cutaneous head and neck squamous cell carcinoma (CHNSCC). METHODS: Between 1977 and 1997, 343 patients presented with CHNSCC, of whom 41 were treated for regional metastases, with parotidectomy and/or (selective) neck dissection. Histological analysis was performed on all operation specimens. Patients with multiple nodes and/or extra capsular rupture, at risk for recurrence, received adjuvant radiotherapy. RESULTS: Seventy-six percent of the regional metastases occurred within the first 2 years, but a delay of more than 5 years was also observed. Parotid gland (56%), neck levels II (39%) and V (22%) were most frequently involved. Twenty-four percent of patients treated with curative intent failed at the regional site. Five years overall survival was 46%, with a median survival of 49 months. No survival differences emerged between patients treated by surgery alone and patients receiving adjuvant radiotherapy (P=0.14). Five patients died of disease, of whom three with distant metastases. CONCLUSION: Regionally metastasized CHNSCC is a serious disease with a high risk of regional recurrence, calling for combined surgery and adjuvant radiotherapy. A small subset of patients with a long interval between completion of primary treatment and presence of regional nodes justifies a long follow-up. PMID- 12559083 TI - The significance of adrenal metastases from lung carcinoma. AB - AIM: The aim of our study was to correlate spread of the lung cancer into the adrenal glands with the progression of the primary disease. METHODS: We diagnosed and confirmed adrenal metastases in 50 patients with non-small cell lung cancer (NSCLC). We correlated the site of the primary lung carcinoma with the site of the adrenal metastasis, and the adrenal metastasis pattern, ipsi-, contra-, and bilateral adrenal metastases, with the operability and number of other sites of metastatic disease. RESULTS: Adrenal metastases were ipsilateral in 20 patients, contralateral in 15 patients and bilateral in 15 patients. An inverse incidence of contra- and bilateral metastasis was observed in 37% of operated patients, and in 71% of patients with inoperable carcinoma. The difference between both groups was statistically significant (P=0.034). CONCLUSIONS: We suggest that an isolated ipsilateral adrenal metastasis in a patient with resectable primary NSCLC could be considered (and treated) as a localized disease rather than a symptom of systemic spread. PMID- 12559084 TI - Quality assurance in surgical oncology. Colorectal cancer as an example. AB - Quality assurance in surgical oncology is a field of growing importance. National, regional and local systems have been built up in many countries. Often the quality assurance projects are linked to different registers. The advantage of such a link is the possibility of obtaining population-based data from unselected health care institutions. Few discussions of results from such projects have been published. Quality assurance of colorectal cancer surgery implies the development and use of systems for improvement all the way from detection of the cancer to the outcome as survival and patient satisfaction. To achieve this we must know what methods are being used and the outcome of our treatments. Designing processes for improvement necessitates careful planning, including decisions about end-points. Some crucial issues are discussed step-by step in the present paper. In addition to auditing and providing collegial feedback, quality assurance is a tool for closing the gap between clinical practice and evidence based medicine and for creating new evidences as well as monitoring the introduction of new techniques and their effects. PMID- 12559085 TI - Lymphatic mapping with tracer administration into the primary breast cancer. AB - There is an ongoing debate over the best tracer injection technique in lymphatic mapping for breast cancer. The technique of low tracer volume administration into the primary breast cancer is presented. The reasons that led to this approach are explained as well as its advantages. Excision of radioactivity that remains at the injection site in the breast cancer prevents the gamma ray scatter that may hamper retrieval of a sentinel node. The intralesional injection technique avoids potential injection of tracer fluid across a lymphatic watershed, it enables identification of extra-axillary sentinel nodes and allows probe-guided excision of non-palpable tumours. PMID- 12559086 TI - Myxoid solitary fibrous tumour of the ischiorectal fossa. AB - Myxoid solitary fibrous tumours are a rare morphological sub-type of the rare solitary fibrous tumours. This case report appears to be the first reported ischiorectal occurrence of this rare tumour. PMID- 12559087 TI - Neurons colocalizing urocortin and cocaine and amphetamine-regulated transcript immunoreactivities are induced by acute lipopolysaccharide stress in the Edinger Westphal nucleus in the rat. AB - Besides corticotropin releasing factor, central stress regulatory pathways utilize various neurotransmitters/neuropeptides, such as urocortin and cocaine and amphetamine-regulated transcript, which play an important role in modifying the efferent components of endocrine, immune and behavioral responses to stress. Urocortin's distribution in the rat's brain has been demonstrated, with the most abundant urocortin-ir perikarya present in Edinger-Westphal nucleus. Cocaine and amphetamine-regulated transcript is widely expressed in the rat brain, with a dominant seat of cellular expression also in the Edinger-Westphal nucleus. Since immediate early gene expressions were seen in several midbrain regions, such as in the Edinger-Westphal nucleus, following various acute stresses, the Edinger Westphal nucleus has been postulated to exert a regulatory/modulatory control over stress responses. Based on these data we decided to investigate the possible colocalization of urocortin and cocaine and amphetamine-regulated transcript-ir in the Edinger-Westphal nucleus using semithin double-label immunofluorescence technique. Furthermore, we also studied whether urocortinergic neurons colocalizing with cocaine and amphetamine-regulated transcript are recruited by lipopolysaccharide stress. Our experiments revealed that urocortin and cocaine and amphetamine-regulated transcript immunoreactivities colocalize in the Edinger Westphal nucleus. In addition, our studies using the inducible immediate early gene c-fos as a marker of activated neurons demonstrated a significant stress induced activation in perikarya colocalizing urocortin- and cocaine and amphetamine-regulated transcript-ir in the Edinger-Westphal nucleus. In view of these data it can be postulated that neurons colocalizing cocaine and amphetamine regulated transcript and urocortin immunoreactivities respond to acute stress, and may play a role in modulating various physiological functions, such as feeding behaviors. PMID- 12559088 TI - Comparative gene identification-94--a pivotal regulator of apoptosis. AB - Apoptosis is an active form of cell death that is carried out by proteins that are designed to kill the cell during normal mammalian development and tissue homeostasis. Cell death by apoptosis comprises a sequence of events leading to the activation of caspases which execute the fragmentation of the cellular protein and DNA leading to disintegration of the cell. This physiological neuronal apoptosis allows the nervous system to eliminate excess neurons. In addition, apoptotic cell death occurs in a variety of neuronal degeneration such as Alzheimer's disease. Here we describe second mitochondria-derived activator of caspases/Diablo as a new interacting protein of CGI-94 (comparative gene identification-94) which itself is probably involved in degenerative processes of Alzheimer's disease. Our findings that CGI-94 interacts with second mitochondria derived activator of caspases/Diablo, inhibits nerve growth factor-induced neurite outgrowth and that its neuronal expression leads to cell death point to its pivotal role in the control of cellular survival. In conclusion, CGI-94 appears to be involved in processes of neuronal degeneration. PMID- 12559089 TI - Rapid inhibitory effects of corticosterone on calcium influx in rat dorsal root ganglion neurons. AB - Corticosterone may nongenomically affect cell functions in addition to its well characterized effects on gene expression. The purpose of this study is to examine if corticosterone has a rapid nongenomic effect on excitability of dorsal root ganglion neurons by using patch-clamp and single-cell Ca(2+) microfluometry techniques. The results show that corticosterone has a dose-dependent rapid inhibitory effect on the voltage-dependent calcium currents in dorsal root ganglion neurons. Moreover, corticosterone inhibits [Ca(2+)](i) elevation induced by 50 mM high K(+) within just 3 s. The inhibitory effects of corticosterone on the voltage-dependent calcium current and high K(+)-induced calcium influx diminish after adding protein kinase C inhibitor or pretreatment with pertussis toxin for 24 h. Our results demonstrate an nongenomic effect of corticosterone on the excitability of dorsal root ganglion neurons and the effect is mediated through a putative pertussis toxin-sensitive G-protein-coupled receptor and activation of protein kinase C. PMID- 12559090 TI - Analysis of purinergic and cholinergic fast synaptic transmission to identified myenteric neurons. AB - Types and projections of neurons that received cholinergic, purinergic and other fast excitatory synaptic inputs in myenteric ganglia of the guinea-pig distal colon were identified using combined electrophysiological recording, application of selective antagonists, marker dye filling via the recording microelectrode, and immunohistochemical characterisation. Fast synaptic inputs were recorded from all major subtypes of uniaxonal neurons including Dogiel type I neurons, filamentous interneurons, circular muscle motor neurons and longitudinal muscle motor neurons. Fast excitatory postsynaptic potentials were completely blocked by the nicotinic receptor antagonists hexamethonium or mecamylamine in 62% of neurons tested and were partially inhibited in the remaining neurons. The P2 purine receptor antagonist, pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid, reduced the amplitudes of fast excitatory postsynaptic potentials in 20% of myenteric neurons. The 5-hydroxytryptamine(3) receptor antagonist granisetron reduced the amplitude of fast excitatory postsynaptic potentials in only one of 15 neurons tested. In five of five neurons tested, the combination of a nicotinic antagonist, pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid, granisetron and 6-cyano-7-nitroquinoxaline-2,3-dione did not completely block the fast excitatory postsynaptic potentials. Immunohistochemical studies of the neurons that had been identified electrophysiologically and morphologically imply that P2X(2) receptors may mediate fast transmission in some neurons, and that other P2X receptor subtypes may also be involved in fast synaptic transmission to myenteric neurons of the guinea-pig distal colon. Neurons with nicotinic and pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid-sensitive fast excitatory postsynaptic potentials were present in both ascending and descending pathways in the distal colon. Thus, neither cholinergic nor mixed cholinergic/purinergic synaptic responses are confined to a particular class of neuron. The results indicate that acetylcholine and ATP are the major fast excitatory neurotransmitters in guinea-pig distal colon myenteric ganglia. PMID- 12559091 TI - Expression of two membrane fusion proteins, synaptosome-associated protein of 25 kDa and vesicle-associated membrane protein, in choroid plexus epithelium. AB - In addition to being the major site of cerebrospinal fluid formation, the choroid plexus epithelium emerges as an important source of polypeptides in the brain. Physiologically regulated release of some polypeptides synthesized by the choroid plexus has been shown. The molecular mechanisms underlying this polypeptide secretion have not been characterized, however. In the present study, synaptosome associated protein of 25 kDa and vesicle-associated membrane protein, two membrane fusion proteins playing a critical role in exocytosis in neurons and endocrine cells, were found to be expressed in the choroid plexus epithelium. It was also shown that in choroidal epithelium, synaptosome-associated protein of 25 kDa and vesicle-associated membrane protein stably interact. Two members of the vesicle-associated membrane protein family, vesicle-associated membrane protein-1 and vesicle-associated membrane protein-2, were expressed in the rat choroid plexus at the messenger RNA and protein level. However, their newly discovered isoforms, vesicle-associated membrane protein-1b and vesicle-associated membrane protein-2b, produced by alternative RNA splicing, were not detected in choroidal tissue. Immunohistochemistry demonstrated that vesicle-associated membrane protein is confined to the cytoplasm of choroidal epithelium, whereas synaptosome associated protein of 25 kDa is associated with plasma membranes, albeit with a varied cellular distribution among species studied. Specifically, in the rat choroid plexus, synaptosome-associated protein of 25 kDa was localized to the basolateral membrane domain of choroidal epithelium and was expressed in small groups of cells. In comparison, in ovine and human choroidal tissues, apical staining for synaptosome-associated protein of 25 kDa was found in the majority of epithelial cells. These species-related differences in cellular synaptosome associated protein of 25 kDa distribution suggested that the synaptosome associated protein of 25 kDa homologue, synaptosome-associated protein of 23 kDa, is also expressed in the rat choroid plexus, which was confirmed by reverse transcriptase polymerase chain reaction. Our findings suggest that synaptosome associated protein of 25 kDa and vesicle-associated membrane protein are involved in secretion of polypeptides from the choroid plexus epithelium. The presence of synaptosome-associated protein of 25 kDa and its homologue as well as multiple isoforms of vesicle-associated membrane protein in choroidal epithelium may play a role in the apical versus basolateral targeting of secretory vesicles. PMID- 12559092 TI - Laminin chain expression suggests that laminin-10 is a major isoform in the mouse hippocampus and is degraded by the tissue plasminogen activator/plasmin protease cascade during excitotoxic injury. AB - Laminins are important components of the extracellular matrix, and participate in neuronal development, survival and regeneration. The tissue plasminogen activator/plasmin extracellular protease cascade and downstream laminin degradation are implicated in excitotoxin-induced neuronal degeneration. To determine which specific laminin chains are involved, we investigated the expression of laminins in the hippocampus, and the cell types expressing them. Reverse transcription-PCR demonstrated that the messenger RNAs for all laminin chains could be detected in the hippocampus. To determine the localization of laminin chain expression, immunostaining was used. This method showed that alpha5, beta1 and gamma1 are most highly expressed in the neuronal cell layers. Immunoblotting confirmed the hippocampal expression of the chains alpha5, beta1 and gamma1, and RNA in situ hybridization showed a neuronal expression pattern of alpha5, beta1 and gamma1. At early time points following intrahippocampal injection of kainate, alpha5, beta1 and gamma1 chain immunoreactivities were lost. In addition, tissue plasminogen activator-deficient mice, which are resistant to kainate-induced neuronal death, show no significant change in laminins alpha5, beta1 and gamma1 after intrahippocampal kainate injection. Taken together, these results suggest that laminin-10 (alpha5-beta1-gamma1) comprises a major neuronal laminin in the mouse hippocampus, and is degraded before neuronal death during excitotoxic injury by the tissue plasminogen activator/plasmin protease cascade. By identifying a neuronal laminin (laminin-10) that participates in neuronal degeneration after excitotoxic injury, this study clarifies the molecular definition of the extracellular matrix in the hippocampus and further defines a pathway for mechanisms of neuronal death. PMID- 12559094 TI - Sex differences in response to kainic acid and estradiol in the hippocampus of newborn rats. AB - Premature and full-term human infants are at considerable risk of excitotoxic mediated brain damage due to hypoxia-ischemia, infection or other trauma. Glutamate receptor activation is a major source of excitoxicity in the adult and developing brain, and the hippocampus is particularly vulnerable to damage. The seven-day-old rat is a widely used model of pediatric brain damage, in large part due to the relative insensitivity of the brain to exogenous glutamate treatment prior to this age. We have reexamined the possible role of glutamate in pediatric brain damage in the newborn rat using kainic acid treatment and attending to the sex of the animal as well as the effects of pretreatment with the gonadal steroid estradiol. Consistent with previous studies, we found no evidence of damage 7 days posttreatment in the CA1 region of the hippocampus in males or females. There was also little to no damage in the CA2/3 or dentate gyrus of males. In females, however, kainic-acid treatment induced substantial damage in the dentate gyrus and moderate damage in CA2/3, as assessed by neuron number and regional volume. Pretreatment with estradiol was protective against kainic acid-induced damage in females but was permissive for damage in the dentate gyrus of males. Estradiol treatment in the absence of kainic acid treatment was also neuroprotective in females in that it increased neuron number and volume throughout the hippocampal formation, suggesting that the basis of the sex difference observed in hippocampal volume was hormonally mediated. There was no effect of exogenous estradiol given to males in the absence of kainic acid. We conclude that the newborn female rat brain, but not the male, is sensitive to glutamate-mediated toxicity and that gonadal steroids play a complex role in both naturally occurring sex differences in hippocampal volume and response to injury. PMID- 12559093 TI - Migration and differentiation of adult rat subventricular zone progenitor cells transplanted into the adult rat striatum. AB - Adult brain subventricular zone progenitor cells undergo neurogenesis in the olfactory bulb. We tested the hypothesis that cultured adult subventricular zone progenitor cells migrate and differentiate into neurons when transplanted into the adult striatum. Cells in the adult rat subventricular zone were isolated and cultured for 8 days in medium containing basic fibroblast growth factor. These cells proliferated as assayed by bromodeoxyuridine immunostaining, and the majority of them were neuron-specific class III beta-tubulin (TuJ1) immunoreactive at 8 days of culture. These cultured cells were labeled in vitro with bromodeoxyuridine or with lipophilic dye-coated particles and were transplanted into the adult rat striatum. Twenty-eight days after transplantation, the cells migrated 0.5-1.5 mm from the midline of the graft to the surrounding host striatum. Migration of grafted cells in the host striatum was also detected on magnetic resonance imaging in living rats. Morphological analysis revealed that many of these migrated cells exhibited multibranched processes from the cell soma resembling host medium-size striatal projection neurons. Only a few astrocyte-like cells were detected. Double immunostaining showed that many bromodeoxyuridine immunoreactive cells were microtubule associated protein 2 or immunoreactive with a mouse monoclonal antibody against neuronal nuclear protein, whereas only a few bromodeoxyuridine immunoreactive cells had glial fibrillary acidic protein immunoreactivity. Morphology of bromodeoxyuridine and microtubule-associated protein 2 immunoreactive cells was similar to those of host microtubule-associated protein 2 immunoreactive cells. These results suggest that transplanted cultured adult subventricular zone progenitor cells can migrate and differentiate in response to guidance cues within the adult striatum. PMID- 12559095 TI - Postnatal development of the hypothalamic neuropeptide Y system. AB - In the adult rat, arcuate-neuropeptide Y/agouti-related protein neurons have efferent projections throughout the hypothalamus and provide a potent orexigenic stimulus. At birth neuropeptide Y fibers are also present throughout the hypothalamus; however, the source of these fibers has been unknown. The present studies determined the postnatal ontogeny of arcuate-neuropeptide Y fibers into the paraventricular nucleus and dorsomedial hypothalamic nucleus, as well as the ontogeny of neuropeptide Y1 receptor expression within these areas. Agouti related protein messenger RNA and protein expression was present exclusively in cell bodies in the arcuate throughout postnatal development, starting at P2, and was colocalized in the vast majority of arcuate-neuropeptide Y neurons. This exclusive colocalization of agouti-related protein with arcuate-neuropeptide Y neurons makes it an excellent marker for these neurons and their projections. Even though single-label neuropeptide Y fibers were abundant in the dorsomedial hypothalamic nucleus and paraventricular nucleus as early as P2, arcuate neuropeptide Y/agouti-related protein fibers did not significantly innervate these areas until P5-6 and P10-11, respectively. In contrast, a portion of the neuropeptide Y fibers within the paraventricular nucleus as early as P2 originated from the brainstem, as indicated by their colocalization with dopamine beta hydroxylase. It remains to be determined if local sources of neuropeptide Y expressing cells within the dorsomedial hypothalamic nucleus and paraventricular nucleus also contribute to the neuropeptide Y-immunoreactive fibers within these regions prior to the development of arcuate-neuropeptide Y/agouti-related protein projections. In addition to the dramatic change in arcuate-neuropeptide Y/agouti related protein projections, there is also a striking change in Y1 protein expression in the hypothalamus during the first two postnatal weeks. Taken together these data suggest that the early postnatal period, during which there is a dynamic change in the hypothalamic neuropeptide Y system, may constitute a critical period in the development of this important feeding circuit. PMID- 12559096 TI - Depolarization waves in the embryonic CNS triggered by multiple sensory inputs and spontaneous activity: optical imaging with a voltage-sensitive dye. AB - Previously, we discovered a novel type of depolarization wave in the embryonic chick brain by using a multiple-site optical recording technique with a fast voltage-sensitive dye. This depolarization wave traveled widely over almost all the region of the CNS. This profile has raised the possibility that the depolarization wave plays some global roles in development of the CNS, rather than contributing to a specific neuronal circuit formation. To obtain more information concerning this issue, in the present study, we examined whether the depolarization wave was triggered by various types of peripheral nerve inputs. Stimulation applied to the vagus, glossopharyngeal, cochlear and trigeminal nerves evoked widely spreading depolarization waves with similar spatiotemporal distribution patterns. The developmental sequence of wave expression was parallel to the development of the excitatory postsynaptic potentials in each sensory nucleus. The depolarization wave was accompanied by a Ca(2+)-wave, suggesting that not only electrical synchrony, but also large-scale Ca(2+)-transients may affect developmental processes in the embryonic brain. Furthermore, we found that the depolarization wave also occurred spontaneously. The waveform and distribution patterns of the spontaneous optical signals were similar to those of the cranial nerve-evoked depolarization wave. These results demonstrated that the depolarization wave in the embryonic chick brain is triggered by multiple sources of external and endogenous activity. This profile supports the idea that this depolarization wave may not serve as a simple regulator of specific neuronal circuit formation, but might play more global roles in CNS development. PMID- 12559097 TI - Chronic lithium treatment antagonizes glutamate-induced decrease of phosphorylated CREB in neurons via reducing protein phosphatase 1 and increasing MEK activities. AB - The cyclic AMP response element binding protein (CREB) has major roles in mediating adaptive responses at glutamatergic synapses and in the neuroprotective effects of neurotrophins. CREB has been implicated as a potential mediator of antidepressant actions. In vitro, chronic lithium treatment has been shown to promote neuronal cell survival. In the present study, we have used cultures of cerebellar granule neurons to analyze the effects of acute and chronic lithium treatment on the response to toxic concentrations of glutamate. Such concentrations of glutamate decrease the phosphorylation of CREB at serine(133) in an N-methyl-D-aspartate (NMDA) receptor-dependent manner. Chronic, but not acute, lithium treatment suppresses glutamate-induced decreases in phosphorylated CREB, and transfection studies indicate that chronic lithium, in the presence of a glutamate stimulus, markedly increases CRE-driven gene expression. Experiments with selected pharmacological reagents indicate that the glutamate-induced decreases in phosphorylated CREB are regulated primarily by protein phosphatase 1. Chronic lithium treatment not only decreases protein phosphatase 1 activity under these circumstances, but also augments glutamate-induced increases in MEK activity. PD 98059, a MEK inhibitor, prevents chronic lithium treatment from increasing phosphorylated CREB levels in glutamate-treated neurons. We conclude from these results that chronic lithium treatment is permissive for maintaining higher phosphorylated CREB levels in the presence of glutamate in part by decreasing protein phosphatase 1 activity and in part by increasing MEK activity. Higher levels of phosphorylated CREB and CRE-responsive genes such as bcl-2 may be responsible for lithium's reported effects on neuronal survival. PMID- 12559098 TI - Protective effect of apolipoprotein E-mimetic peptides on N-methyl-D-aspartate excitotoxicity in primary rat neuronal-glial cell cultures. AB - Apolipoprotein E (apoE) is a 34-kD protein with multiple biological properties. Recent clinical and preclinical observations implicate a role for apoE in modifying the response of the brain to focal and global ischemia. One mechanism by which apoE might exert these effects is by reducing glutamate-induced excitotoxic neuronal injury associated with ischemic insults. We demonstrate that human recombinant apoE confers a mild neuroprotective effect in primary neuronal glial cultures exposed to 100 microM N-methyl-D-aspartate. Furthermore, a peptide derived from the receptor-binding region of apoE (residues 133-149) maintained a significant helical population as assessed by circular dichroism, and completely suppressed the neuronal cell death and calcium influx associated with N-methyl-D aspartate exposure. Neuroprotection was greatest when the peptide was added concurrently with N-methyl-D-aspartate; however, a significant protection was observed when peptide was preincubated and washed off prior to N-methyl-D aspartate exposure. These results suggest that one mechanism by which apoE may modify the CNS response to ischemia is by partially blocking glutamate excitotoxicity. Moreover, small peptide fragments derived from the receptor binding region of apoE have enhanced bioactivity compared with the intact holoprotein, and may represent a novel therapeutic strategy for the treatment of brain ischemia. PMID- 12559099 TI - Investigations of the cholinergic modulation of GABA release in rat thalamus slices. AB - The thalamus receives a dense cholinergic projection from the pedunculopontine tegmentum. A number of physiological studies have demonstrated that this projection causes a dramatic change in thalamic activity during the transition from sleep to wakefulness. Previous anatomical investigations have found that muscarinic type 2 receptors are densely distributed on the dendritic terminals of GABAergic interneurons, as well as the somata and proximal dendrites of GABAergic cells in the thalamic reticular nucleus. Since these structures are the synaptic targets of cholinergic terminals in the thalamus, it appears likely that thalamic pedunculopontine tegmentum terminals can activate muscarinic type 2 receptors on GABAergic cells. To test whether activation of muscarinic type 2 receptors affects the release of GABA in the thalamus, we have begun pharmacological studies using slices prepared from the rat thalamus. We have found that the application of the nonspecific muscarinic agonist, methacholine, and the muscarinic type 2-selective agonist, oxotremorine.sesquifumarate, diminished both the baseline, and K(+) triggered release of [(3)H]GABA from thalamic slices. This effect was calcium dependent, and blocked by the nonselective muscarinic antagonist atropine, the muscarinic type 2-selective antagonist, methoctramine, but not the muscarinic type 1 antagonist, pirenzepine. Thus, it appears that one function of the pedunculopontine tegmentum projection is to decrease the release of GABA through activation of muscarinic type 2 receptors. This decrease in inhibition may play an important role in regulating thalamic activity during changes in states of arousal. PMID- 12559100 TI - Maintenance of sympathetic tone by a nickel chloride-sensitive mechanism in the rostral ventrolateral medulla of the adult rat. AB - In urethane-anaesthetised artificially ventilated Sprague-Dawley rats, bilateral microinjection of the divalent cation nickel chloride (Ni(2+); 50 mM, 50 nl) into the rostral ventrolateral medulla elicited a dramatic inhibition of splanchnic sympathetic nerve activity (-44+/-6%) and a marked depressor response (-35+/-7 mmHg). Selective blockade of high-voltage activated Ca(2+) channels with omega agatoxin IVA (P/Q-type), omega-conotoxin GVIA (N-type) and nifedipine (L-type) did not decrease arterial pressure or splanchnic sympathetic nerve activity when injected separately into the rostral ventrolateral medulla, or combined with kynurenate. Injection of caesium chloride or ZD 7288, a blocker of the hyperpolarization-activated cation current, into the rostral ventrolateral medulla had no effect on arterial pressure or splanchnic sympathetic nerve activity. Bilateral microinjection of nickel chloride into the caudal ventrolateral medulla/pre-Botzinger complex elicited small increases in splanchnic sympathetic nerve activity (+17+/-13%) and arterial pressure (+12+/-4 mmHg). These were substantially smaller than those evoked by blockade of glutamatergic receptors or high-voltage activated Ca(2+) channels in this area. Injection of kynurenate or high-voltage activated Ca(2+) channel blocker, but not Ni(2+), in this area evoked respiratory termination. The results indicate the existence of a distinct mechanism maintaining the tonic activity of rostral ventrolateral medulla presympathetic neurons that is different from that maintaining the tonic activity in the caudal ventrolateral medulla/pre-Botzinger region. We conclude that ion channels that are sensitive to Ni(2+), but are insensitive to high-voltage activated (L, P/Q, N) Ca(2+) channel blockers, and are located postsynaptically on the presympathetic rostral ventrolateral medulla neurons are responsible for the tonic activity of the presympathetic neurons in rostral ventrolateral medulla. These channels could well be the low-voltage activated (or T-type) Ca(2+) channels although other conductances cannot be conclusively excluded. PMID- 12559101 TI - Effects of fructose-1,6-bisphosphate on morphological and functional neuronal integrity in rat hippocampal slices during energy deprivation. AB - D-fructose-1,6-bisphosphate, a high energy glycolytic intermediate, attenuates ischemic damage in a variety of tissues, including brain. To determine whether D fructose-1,6-bisphosphate serves as an alternate energy substrate in the CNS, rat hippocampal slices were treated with D-fructose-1,6-bisphosphate during glucose deprivation. Unlike pyruvate, an endproduct of glycolysis, 10 mM D-fructose-1,6 bisphosphate did not preserve synaptic transmission or morphological integrity of CA1 pyramidal neurons during glucose deprivation. Moreover, during glucose deprivation, 10-mM D-fructose-1,6-bisphosphate failed to maintain adenosine triphosphate levels in slices. D-fructose-1,6-bisphosphate, however, attenuated acute neuronal degeneration produced by 200 microM iodoacetate, an inhibitor of glycolysis downstream of D-fructose-1,6-bisphosphate. Because (5S, 10R)-(+)-5 methyl-10, 11-dihydro-5H-dibenzo [a,d]cyclohepten-5,10-imine, an antagonist of N methyl-D-aspartate receptors, exhibited similar protection against iodoacetate damage, we examined whether (5S, 10R)-(+)-5-methyl-10, 11-dihydro-5H-dibenzo [a,d]cyclohepten-5,10-imine and D-fructose-1,6-bisphosphate share a common neuroprotective mechanism. Indeed, D-fructose-1,6-bisphosphate diminished N methyl-D-aspartate receptor-mediated synaptic responses and partially attenuated neuronal degeneration induced by 100-microM N-methyl-D-aspartate. Taken together, these results indicate that D-fructose-1,6-bisphosphate is unlikely to serve as an energy substrate in the hippocampus, and that neuroprotective effects of D fructose-1,6-bisphosphate are mediated by mechanisms other than anaerobic energy supply. PMID- 12559102 TI - Role of protein kinase C in central muscarinic inhibitory mechanisms regulating voiding in rats. AB - To evaluate the role of protein kinase C in central muscarinic mechanisms regulating voiding, cystometry was performed in conscious rats. Oxotremorine methiodide, a muscarinic agonist was injected i.c.v. in a dose (0.1 microg/rat) shown previously to alter voiding function. Oxotremorine methiodide was also tested after i.c.v. injection of chelerythrine chloride (a protein kinase C inhibitor, 2 microg/rat) or 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7, a protein kinase inhibitor, 5 nmol/rat). In untreated rats, oxotremorine methiodide elicited a bimodal response consisting of an initial increase in bladder capacity, maximal voiding pressure, pressure threshold and post voiding intravesical pressure, but reduced voiding efficiency and bladder compliance. The second response consisted of a decrease in bladder capacity and bladder compliance, increases in maximal voiding pressure and post voiding intravesical pressure, but no change in pressure threshold or voiding efficiency. However, approximately 20 min after pre-treatment with chelerythrine chloride or H-7 in doses that did not alter voiding function, oxotremorine methiodide decreased bladder capacity, increased maximal voiding pressure, but did not change pressure threshold or voiding efficiency. These results indicate that inhibitory and facilitatory muscarinic mechanisms in the brain that control voiding function involve different second messenger systems. Inhibitory mechanisms which are blocked by chelerythrine chloride or H-7 must involve protein kinase C and normally be inactive because the protein kinase inhibitors alone did not alter voiding. On the other hand, facilitatory muscarinic mechanisms which previous studies showed were tonically active are not mediated by chelerythrine chloride or H-7 sensitive signaling pathways. PMID- 12559103 TI - Valproate inhibits oxidative damage to lipid and protein in primary cultured rat cerebrocortical cells. AB - Valproate is often prescribed as a long-term therapeutic mood stabilizing agent for individuals with bipolar disorder. Although research suggests that this drug may produce a neuroprotective effect, its neuroprotective mechanism is not yet clear. The purpose of this study was to determine if valproate provides a neuroprotective effect against damage caused by oxidative stress in primary cultured rat cerebral cortical cells. We found that chronic treatment with valproate at therapeutically relevant concentrations for 7 days inhibited lipid peroxidation and protein oxidation induced by treatment with 0.25 mM oxidant FeCl(3) for 90 min, indicating that valproate inhibits oxidative damage to lipid and protein. Our results suggest that chronic treatment with valproate may protect neuronal cells from damage caused by oxidative stress and that neuroprotection from oxidative damages may be involved in the mechanism of action of valproate. Supporting this possibility are recent findings that chronic treatment with valproate increased the expression of endoplasmic reticulum stress protein GRP78 and antiapoptotic factor bcl-2 in rat cerebral cortex. Since GRP78 binds Ca(2+) and folds damaged protein, bcl-2 stabilizes mitochondrial transmembrane potential and inhibits cytochrome C release, and both GRP78 and bcl 2 have been shown to inhibit oxyradical accumulation, together these findings indicate that valproate may target one or more of these processes in order to produce neuroprotective effects. PMID- 12559104 TI - Gonadal hormones act extrinsic to the hippocampus to influence the density of hippocampal astroglial processes. AB - The important effects of estrogen on the morphology of hippocampal neurons are well established. The mechanisms leading to such changes, nevertheless, have proved confusingly complex, since interactions between glia and neurons, as well as neuronal influences from other brain fields, are involved. This study addresses the possibility that estrogen-sensitive projections from the medial septum/diagonal band of Broca induce astroglial reactions. Estrogen- and cholesterol-filled (controls) cannulae were implanted into the medial septum/diagonal band of Broca of adult ovariectomized rats. Comparative semiquantitative immunohistochemical analysis on the density of the glial fibrillary acidic protein-containing processes and cells were performed on hippocampal slices of locally estrogen-treated and control animals. Rats that received estrogen-filled cannulae showed a lower density of glial processes in the hippocampal CA1 and CA3 subfields than animals of the control group. These effects could not be observed in the dentate gyrus. Cell counts revealed no significant difference in the number of glial fibrillary acidic protein-positive cells in any of the examined areas. Two major conclusions can be drawn from these results. First, the data show that estrogen, in fact, has an indirect influence on hippocampal cells through septo-hippocampal projections. Furthermore, estradiol can have an indirect negative effect on hippocampal astrocytes, causing a reduction in the density of their processes. PMID- 12559105 TI - Differences in lesion-induced hippocampal plasticity between mice and rats. AB - We studied the differences between mice and rats in lesion-induced sprouting in the hippocampus. The entorhinal cortex was unilaterally lesioned with ibotenic acid in adult, female mice and rats. Four weeks later the subsequent axonal sprouting in the dentate gyrus was analysed, by measuring the density of the synaptophysin immunohistochemical and acetylcholinesterase histochemical staining in the termination area of the entorhinal cortex axons. The data demonstrate that both mice and rats display a significantly increased density of staining for synaptophysin and acetylcholinesterase in the molecular layer of the dentate gyrus, indicative of axonal sprouting. Both species also show an upregulation in the density of staining for acetylcholinesterase in the molecular layer of the dentate gyrus. Further, rats, but not mice, show a significant upregulation of synaptophysin staining in stratum lacunosum moleculare of CA1 following the lesions. However, whereas rats show significant shrinkage of the molecular layer of the dentate gyrus, mice do not show any shrinkage of that layer following entorhinal cortex lesions. Taken together, these data indicate that whereas the process of reinnervation in the hippocampus is similar between the mouse and the rat, the hippocampal response to denervation shows clear differences between these two species. PMID- 12559106 TI - Altered auditory-evoked potentials in mice carrying mutated human amyloid precursor protein and presenilin-1 transgenes. AB - Transgenic mice carrying human APPswe and PS1-A264E transgenes (A/P mice) have elevated levels of the highly fibrillogenic amyloid Abeta(1-42) (Abeta) and develop amyloid plaques around the age of 9 months. Our aim was to find whether the gradual accumulation of Abeta in these mice can be detected with long-term recording of auditory-evoked potentials. The A/P double-mutant mice had impaired auditory gating and a tendency toward increased latency of the cortical N35 response, but these changes were not age-dependent between 7 and 11 months of age. In a control experiment that included also APP and PS1 single-mutant mice, the A/P double-mutant mice had weaker auditory gating than either APP or PS1 mice. In contrast, increased N35 latency was found in both A/P and APP mice compared with nontransgenic or PS1 mice. The Abeta40 and Abeta42 levels were robustly increased in A/P mice and Abeta40 moderately increased also in APP mice. Plaques were deposited only in A/P mice. We conclude that the impaired auditory gating is associated with the overproduction Abeta42 but does not reflect its amount. In contrast, increased N35 latency is related to the APP genotype independent of Abeta42 production. PMID- 12559107 TI - Corticotropin-releasing factor as well as opioid and dopamine are involved in tail-pinch-induced food intake of rats. AB - Several kinds of stress such as psychological stress, restraint, and foot shock inhibit feeding behavior through corticotropin-releasing factor (CRF). In contrast, a mild tail pinch increases food intake in rats. Although dopamine and opioid are thought to be involved in tail-pinch-induced food intake, it is unknown whether CRF participates in this phenomenon. Therefore, we attempted to clarify this issue using rats. A 30-s tail pinch increased food intake in 30 min after the tail pinch, and this increase was blocked by intraperitoneal injection of CRF receptor type 1 selective antagonist. CRF increased food intake in 30 min after intracerebroventricular injection at a dose of 2 or 10 ng, and this increase was also blocked by CRF receptor type 1 antagonist. Tail-pinch- or CRF induced food intake was blocked by naloxone, pimozide, and spiperone. These results suggest that CRF, through CRF receptor type 1 as well as opioid and dopaminergic systems, are involved in the mechanism of tail-pinch-induced food intake. The results also suggest that brain CRF has dual effects on food intake, hyperphagia and anorexia, in a stress-dependent manner. PMID- 12559108 TI - Fluctuations in neural activity during cocaine self-administration: clues provided by brain thermorecording. AB - Since metabolic neural activity is accompanied by heat release, measurement of local brain temperature offers a method for assessing alterations in neural activity. This approach, continuous monitoring of local brain (ventral tegmental area, ventral striatum, and hippocampus) and body (temporal muscle) temperature, was used to study intravenous cocaine self-administration in trained rats. The first self-administration of a session was preceded by a strong temperature increase that continued after the drug infusion. After peaking at the time of the second self-administration, temperature plateaued (+0.7 degrees C) with biphasic fluctuations (+/-0.10-0.15 degrees C) around each subsequent self-administration. Temperature gradually increased before and for 30-50 s after the lever-press, but then abruptly decreased to a minimum at 180-240 s, when it began to increase to reach another peak immediately after the next lever-press. Doubling the dose of injected cocaine significantly potentiated the post-cocaine temperature decrease and increased time to the next lever-press. In contrast to drug-reinforced lever presses, temperatures phasically increased after non-reinforced lever-presses and at the end of a session when the lever was blocked and the rat was hyperactive, trying to reach the inaccessible lever. While temperature changes in each recording location were generally correlative, the initial temperature elevation was stronger in all brain structures than in muscle and ventral striatum was the structure that showed the most pronounced and consistent temperature fluctuations. These data suggest a generalized brain activation associated with cocaine-seeking and cocaine-taking behavior with its phasic fluctuations around individual drug self-injections. While the initial component of brain activation preceding the first lever-press for cocaine is internally determined and closely related to behavioral search, subsequent biphasic fluctuations in neural activity associated with repeated drug intakes appear to be drug-mediated. Cocaine-induced potentiation of monoamine transmission is a possible factor for gradual increases in neural activity that drive cocaine seeking, while a rapid, brain concentration dependent action on Na(+) transport (local anesthetic action) is the most probable factor determining an abrupt, transient cessation of neural activation associated with cocaine reward. PMID- 12559109 TI - Changes in brain cholecystokinin and anxiety-like behavior following exposure of mice to predator odor. AB - Exposure of CD-1 mice to a familiar environment lined with clean shavings (control odor) as well as a familiar environment lined with soiled rat shavings (predator odor) induced anxiety in the light/dark box. Mice exposed to the familiar environment or predator odor displayed decreased latency to enter the dark chamber of the light/dark box and spent less time in the light portion of the apparatus relative to home-caged mice. Mice exposed to the familiar environment lined with clean shavings or predator odor displayed elevated cholecystokinin mRNA levels from the ventral tegmental area, medial and basolateral nuclei of the amygdala relative to home-caged mice. Exposure of CD-1 mice to 2, 5 or 10 min of predator odor increased acoustic startle relative to mice merely exposed to the familiar environment lined with clean shavings at protracted intervals. Mice exposed to the familiar environment lined with clean shavings did not exhibit enhanced startle relative to home-caged mice. Exaggerated startle reactivity was in evidence immediately, 24, and 48 h following a 5-min exposure of mice to predator odor. In contrast, a 10-min exposure of mice to predator odor produced an oscillating pattern of enhanced startle evident during the immediate and 48-h post-stressor intervals only. However, when the startle stimulus was withheld 1 h following odor presentation, mice exhibited enhanced startle patterns reminiscent of the 5-min exposure. The 2 min exposure of mice to predator odor produced a delayed onset of enhanced startle observed at the 168-h test interval only. Potential anxiogenic influences of mesocorticolimbic cholecystokinin availability as well as the time course and underlying neuronal substrates of long-term behavioral disturbances as a result of psychogenic stressor manipulations are discussed. PMID- 12559110 TI - Temporal patterns of limbic monoamine and plasma corticosterone response during social stress. AB - Dominant and subordinate males respond differently to the stress of social interaction. After an hour of social interaction, subordinate male Anolis carolinensis have elevated serotonergic activity in hippocampus, but dominant males do not. In other species, and using other stressors, the activation of hippocampal serotonergic activity is much more rapid than one hour. To elucidate early stress responsiveness, adult male A. carolinensis were divided into four groups: isolated controls, and pairs of males sampled after 10, 20 or 40 minutes of aggressive interaction. Development of dominant-subordinate relationships was determined by behavior and by the celerity of eyespot darkening. Serotonergic activity in the hippocampus, nucleus accumbens and amygdala was elevated rapidly and equally in both dominant and subordinate males, as were plasma corticosterone concentrations. Serotonergic activity remained elevated through 40 minutes in hippocampus and nucleus accumbens. Only subordinate males had elevated corticosterone levels at 40 minutes. Social status does not impede socially induced stress responses. Rather, rapid regulation of serotonergic stress responses appears to be a mediating factor in determining both behavioral output and social status. Temporal expressions of monoaminergic and endocrine stress responses are distinctive between males of dominant and subordinate social status. Such temporal patterns of transmitter and glucocorticoid activity may reflect neurocircuitry adaptations that result in behavior modified to fit social status. PMID- 12559111 TI - Postsynaptic dorsal column neurons express NK1 receptors following colon inflammation. AB - Recent clinical and experimental studies have suggested that the dorsal column pathway and specifically postsynaptic dorsal column neurons play an important role in the transmission of visceral pain. In our study we have mapped the distribution of postsynaptic dorsal column neurons in thoracic, lumbar and sacral spinal cord segments. The presence of immunoreactivity for neurokinin 1 receptors on these postsynaptic dorsal column neurons was examined under control conditions and after colon inflammation. The largest number of postsynaptic dorsal column neurons was found in the lumbar enlargement. They were mostly located in laminae III-IV, but depending on the spinal segment, about 7-15% of them were in the deep medial dorsal horn and in the central canal area. Under control conditions none of the 1438 postsynaptic dorsal column neurons examined expressed neurokinin 1 receptors. However, after induction of colon inflammation about 1.4% of the 2015 postsynaptic dorsal column neurons observed in the experimental group showed immunoreactivity for neurokinin 1 receptors. These neurons were preferentially found in the lower thoracic and lumbosacral spinal segments where they represented about 3-4% of the total population of postsynaptic dorsal column neurons examined. The de novo expression of neurokinin1 receptors on postsynaptic dorsal column neurons after colon inflammation suggests that substance P released from visceral primary afferents under inflammatory conditions could help produce central sensitization of these neurons. PMID- 12559112 TI - Mu-opioid receptors are located postsynaptically and endomorphin-1 inhibits voltage-gated calcium currents in premotor cardiac parasympathetic neurons in the rat nucleus ambiguus. AB - Activation of opioid receptors in the CNS evokes a dramatic decrease in heart rate which is mediated by increases in inhibitory parasympathetic activity to the heart. Injection of opiates into the nucleus ambiguus, where premotor cardiac parasympathetic nucleus ambiguus neurons are located elicits an increase in parasympathetic cardiac activity and bradycardia. However, the mechanisms responsible for altering the activity of premotor cardiac parasympathetic nucleus ambiguus neurons is unknown. This study examined at the electron microscopic level whether premotor cardiac parasympathetic nucleus ambiguus neurons possess postsynaptic opioid receptors and whether mu-opioid receptor agonists alter voltage-gated calcium currents in these neurons. Premotor cardiac parasympathetic nucleus ambiguus neurons were identified in the rat using retrograde fluorescent tracers. One series of experiments utilized dual-labeling immunocytochemical methods combined with electron microscopic analysis to determine if premotor cardiac parasympathetic nucleus ambiguus neurons contain mu-opioid receptors. In a second series of experiments whole cell patch clamp methodologies were used to determine whether activation of postsynaptic opioid receptors altered voltage gated calcium currents in premotor cardiac parasympathetic nucleus ambiguus neurons in brainstem slices. The perikarya and 78% of the dendrites of premotor cardiac parasympathetic nucleus ambiguus neurons contain mu-opioid receptors. Voltage-gated calcium currents in premotor cardiac parasympathetic nucleus ambiguus neurons were comprised nearly entirely of omega-agatoxin-sensitive P/Q type voltage-gated calcium currents. Activation of mu-opioid receptors inhibited these voltage-gated calcium currents and this inhibition was blocked by pretreatment with pertusis toxin. The mu-opioid receptor agonist endomorphin-1, but not the mu-opioid receptor agonist endomorphin-2, inhibited the calcium currents. In summary, mu-opioid receptors are located postsynaptically on premotor cardiac parasympathetic nucleus ambiguus neurons. The mu-opioid receptor agonist endomorphin1 inhibited the omega-agatoxin-sensitive P/Q-type voltage gated calcium currents in premotor cardiac vagal nucleus ambiguus neurons. This inhibition is mediated via a G-protein mediated pathway which was blocked by pretreatment with pertusis toxin. It is possible that the inhibition of calcium currents may act to indirectly facilitate the activity of premotor cardiac parasympathetic nucleus ambiguus neurons by disinhibition, such as by a reduction in inhibitory calcium activated potassium currents. PMID- 12559113 TI - Circadian profile and photic regulation of clock genes in the suprachiasmatic nucleus of a diurnal mammal Arvicanthis ansorgei. AB - The molecular mechanisms of the mammalian circadian clock located in the suprachiasmatic nucleus have been essentially studied in nocturnal species. Currently, it is not clear if the clockwork and the synchronizing mechanisms are similar between diurnal and nocturnal species. Here we investigated in a day active rodent Arvicanthis ansorgei, some of the molecular mechanisms that participate in the generation of circadian rhythmicity and processing of photic signals. In situ hybridization was used to characterize circadian profiles of expression of Per1, Per2, Cry2 and Bmal1 in the suprachiasmatic nucleus of A. ansorgei housed in constant dim red light. All the clock genes studied showed a circadian expression. Per1 and Per2 mRNA increased during the subjective day and decreased during the subjective night. Also, Bmal1 exhibited a circadian expression, but in anti-phase to that of Per1. The expression of Cry2 displayed a circadian pattern, increasing during the late subjective day and decreasing during the late subjective night. We also obtained the phase responses to light for wheel-running rhythm and clock gene expression. At a behavioral level, light was able to induce phase shifts only during the subjective night, like in other diurnal and nocturnal species. At a molecular level, light pulse exposure during the night led to an up-regulation of Per1 and Per2 concomitant with a down regulation of Cry2 in the suprachiasmatic nucleus of A. ansorgei. In contrast, Bmal1 expression was not affected by light pulses at the circadian times investigated. This study demonstrates that light exposure during the subjective night has opposite effects on the expression of the clock genes Per1 and Per2 compared with that of Cry2. These differential effects can participate in photic resetting of the circadian clock. Our data also indicate that the molecular mechanisms underlying circadian rhythmicity and photic synchronization share clear similarities between diurnal and nocturnal mammals. PMID- 12559114 TI - Sex differences in the magnocellular subdivision of the medial preoptic nucleus in Syrian hamsters. AB - Exposure to pheromonal cues initiates male mating behavior. Pheromones are processed within a pathway that converges on the magnocellular subdivision of the medial preoptic nucleus. Lesions of this area eliminate male copulatory behavior, but do not affect anogenital investigation. Exposure to pheromones stimulates cells of the magnocellular subdivision of the medial preoptic nucleus in a sex specific manner. In this study, we hypothesize that sex differences in cell number may underlie sex differences in pheromone-induced neural stimulation. The current study used unbiased stereological methods to identify sexual dimorphisms in the magnocellular subdivision of the medial preoptic nucleus. Sex differences were found in the number and density of neurons, but not in overall volume or neuron volume. Consequently, the total volume is not sexually differentiated because neurons are more densely packed within the male magnocellular subdivision of the medial preoptic nucleus. These results support the hypothesis that additional neurons in the magnocellular subdivision of the medial preoptic nucleus are critical for the expression of male copulatory behaviors in adulthood. Furthermore, they suggest that sexual differentiation of the magnocellular subdivision of the medial preoptic nucleus is an important process that forms the anatomical basis for sex-specific behavioral responses to pheromonal stimulation. PMID- 12559115 TI - Increased dendritic excitability in hippocampal ca1 in vivo in the kainic acid model of temporal lobe epilepsy: a study using current source density analysis. AB - We used kainic acid in rats as an animal model of temporal lobe epilepsy, and studied the synaptic transmission in hippocampal subfield CA1 of urethane anesthetized rats in vivo. Dendritic currents were revealed by field potential mapping, using a single micropipette or a 16-channel silicon probe, followed by current source density analysis. We found that the population excitatory postsynaptic potentials in the basal dendrites and distal apical dendrites of CA1 were increased in kainate-treated as compared with control rats following paired pulse, but not single-pulse, stimulation of CA3b or medial perforant path. In contrast, the trisynaptic midapical dendritic response in CA1 following medial perforant path stimulation was decreased in kainate-treated as compared with control rats. Increased coupling between excitatory postsynaptic potential and the population spike in CA1 was found after kainate seizures. Short-latency, presumably monosynaptic CA1 population spikes following medial perforant path stimulation was found in kainate-treated but not control rats. An enhancement of dendritic excitability was evidenced by population spikes that invaded into or originated from the distal apical dendrites of CA1 in kainate-treated but not control rats. Reverberation of hippocampo-entorhinal activity was evidenced by recurrent excitation of CA1 following CA3b stimulation in kainate-treated but not control rats. Blockade of inhibition by intraventricularly administered bicuculline induced excitatory potentials in CA1 that were stronger and more prolonged in kainate-treated than control rats. The bicuculline-induced excitation was mainly blocked by non-N-methyl-D-aspartate receptor antagonists. We conclude that kainate seizures induced disinhibition in CA1 that unveiled excitation at the basal and distal apical dendrites, resulting in enhancement of the direct entorhinal cortex to CA1 input and reverberations via the hippocampo entorhinal loop. These changes in the output of the hippocampus from CA1 are likely detrimental to the behavioral functions of the hippocampus and they may contribute to increased seizure susceptibility after kainate seizures. PMID- 12559116 TI - Pharmacology of neuronal background potassium channels. AB - Background or leak conductances are a major determinant of membrane resting potential and input resistance, two key components of neuronal excitability. The primary structure of the background K(+) channels has been elucidated. They form a family of channels that are molecularly and functionally divergent from the voltage-gated K(+) channels and inward rectifier K(+) channels. In the nervous system, the main representatives of this family are the TASK and TREK channels. They are relatively insensitive to the broad-spectrum K(+) channel blockers tetraethylammonium (TEA), 4-aminopyridine (4-AP), Cs(+), and Ba(2+). They display very little time- or voltage-dependence. Open at rest, they are involved in the maintenance of the resting membrane potential in somatic motoneurones, brainstem respiratory and chemoreceptor neurones, and cerebellar granule cells. TASK and TREK channels are also the targets of many physiological stimuli, including intracellular and extracellular pH and temperature variations, hypoxia, bioactive lipids, and neurotransmitter modulation. Integration of these different signals has major effects on neuronal excitability. Activation of some of these channels by volatile anaesthetics and by other neuroprotective agents, such as riluzole and unsaturated fatty acids, illustrates how the neuronal background K(+) conductances are attractive targets for the development of new drugs. PMID- 12559118 TI - An increased expression of the mGlu5 receptor protein following LTP induction at the perforant path-dentate gyrus synapse in freely moving rats. AB - The involvement of metabotropic glutamate (mGlu) receptors in the induction of long-term potentiation (LTP) in vivo has been consistently documented. We have investigated whether LTP induction in the dentate gyrus of rats leads to changes in expression of mGlu2/3 or -5 receptor subtypes in the hippocampus. LTP was induced at the medial perforant path-dentate gyrus synapses, and mGlu receptor expression was examined by Western blot or in situ hybridization. An up regulation of mGlu5 receptors was observed in the hippocampus both 24 and 48 h following LTP induction. This effect was restricted to the dentate gyrus and CA1 region, whereas no changes in mGlu5 receptor protein (but an increase in mRNA levels) were observed in the CA3 region. The increased expression of mGlu5 receptors was directly related to the induction of LTP, because it was not observed when tetanic stimulation was carried out in animals treated with the NMDA receptor antagonist, 2-amino-5-phosphonopentanoate (AP5). Western blot analysis also showed a reduced expression of mGlu2/3 receptors in the whole hippocampus 24 h after LTP induction, indicating that the increased expression of mGlu5 receptors was specific. These data suggest that an up-regulation of mGlu5 receptors is a component of the plastic changes that follow the induction of LTP at the perforant path-dentate gyrus synapse. PMID- 12559117 TI - Corticostriatal LTP requires combined mGluR1 and mGluR5 activation. AB - Metabotropic glutamate receptors (mGluRs) have been demonstrated to play a role in synaptic plasticity. It has been recently shown that mGluR1 is involved in corticostriatal long-term depression, by means of pharmacological approach and by using mGluR1-knockout mice. Here, we report that both mGluR1 and mGluR5 are involved in corticostriatal long-term potentiation (LTP). In particular, the mGluR1 antagonist LY 367385, as well as the mGluR5 antagonist MPEP, reduce LTP amplitude. Moreover, blockade of both mGluR1 and mGluR5 by LY 367385 and MPEP co administration fully suppresses LTP. Accordingly, group II and group III mGluRs antagonists fail to affect LTP induction. Interestingly, LTP amplitude is also significantly reduced in both mGluR1- and mGluR5-knockout mice. The differential function of mGluR1 and mGluR5 in corticostriatal synaptic plasticity may play a role in the modulation of the motor activity mediated by the basal ganglia, thus providing a substrate for the pharmacological treatment of motor disorders involving the striatum. PMID- 12559119 TI - Taurine-induced synaptic potentiation and the late phase of long-term potentiation are related mechanistically. AB - The application of taurine (2-aminoethanesulfonic acid) induces a long-lasting increase of synaptic efficacy and axon excitability (LLP-TAU) in rat hippocampal CA1 area. After taurine withdrawal, LLP-TAU lasted at least 3 h. This fact prompted us to assess whether the mechanisms involved in the maintenance of this particular potentiation were similar to those implicated in the late phase of long-term potentiation (L-LTP). In the presence of KN-62, an inhibitor of calcium/calmodulin-dependent protein kinase, taurine perfusion (10 mM, 30 min) did not affect the induction of LLP-TAU. However, LLP-TAU maintenance was completely suppressed by KT5720, an inhibitor of the cAMP-dependent protein kinase (PKA). Moreover, the late phase of LLP-TAU was blocked by inhibiting protein synthesis with anisomycin. In addition, taurine perfusion increased the phosphorylation of cAMP response element-binding protein (CREB), although did not affect cAMP levels. These features of LLP-TAU do not appear to be caused by the activation of D1/D5 dopamine receptors, as taurine also induced synaptic potentiation in the presence of SCH23390, an antagonist of this type of receptors. Finally, the late phase of both L-LTP and LLP-TAU occluded mutually. These results suggest that taurine triggers the sequence of some of the molecular events involved in the induction of L-LTP. PMID- 12559120 TI - Galanin regulates spatial memory but not visual recognition memory or synaptic plasticity in perirhinal cortex. AB - It has previously been shown that the neuropeptide galanin plays a role in the age-dependent regulation of hippocampal synaptic plasticity and spatial memory. Here, we further extend these studies by demonstrating that galanin knockout mice also have deficits in an object-in-place spatial memory task. In contrast however, there is no deficit in single item object recognition memory, a memory that depends on perirhinal cortex. Furthermore, in perirhinal cortex slices there are no differences in activity-dependent long-term potentiation or depotentiation, nor in muscarinic receptor-dependent long-term depression between galanin knockout mice and wild-type litter-mates. Therefore, these results suggest that galanin has a differential role in hippocampal-dependent and perirhinal cortex-dependent memory. PMID- 12559121 TI - On the putative physiological role of allopregnanolone on GABA(A) receptor function. AB - To obtain definitive evidence for a physiological allosteric modulatory role for endogenous brain ALLO on GABA(A) receptor function, we studied GABA(A) receptor activity under conditions in which the concentration of endogenous brain ALLO was decreased by about 80% for longer than 5 h following the administration of SKF 105111- 17beta-17-[bis (1methylethyl) amino carbonyl] androstane-3,5-diene-3 carboxylic acid (SKF), a potent inhibitor of 5alpha-reductases Type I and II. We used the in situ patch-clamp technique to record GABA-evoked currents and spontaneous inhibitory postsynaptic currents (sIPSCs) from pyramidal neurons in neocortical slices of vehicle- or SKF-treated mice. The potency, but not the efficacy, of exogenously applied GABA was decreased in slices from mice treated with SKF. When neocortical slices were treated in vitro for 3 h with 10 microM SKF, ALLO was also reduced (25-30%) and in addition, the GABA dose-response curve was shifted to the right; however this shift was not as marked as the shift in the slices obtained from mice treated with SKF, in keeping with the smaller decrease of the ALLO content in these slices. Furthermore, direct application of ALLO to these slices shifted the dose-response curve of GABA back toward a non SKF treated profile. We then analyzed GABAergic sIPSCs in neocortical slices obtained from vehicle or SKF-treated mice. Mean decay time and charge transfer were significantly reduced by SKF treatment. The decay of sIPSCs was best fitted by two exponentials, but only the fast component was decreased in the SKF group. Direct application of ALLO (100 nM) normalizes the sIPSC kinetics in slices from ALLO depleted mice. No changes were detected in the amplitude or frequency of sIPSCs. These data demonstrate that endogenous ALLO physiologically regulates spontaneously induced Cl(-) current by acting on a specific recognition site, which is probably located on GABA(A) receptors (a receptor on a receptor), thereby prolonging inhibitory currents by facilitating conformational transition of the GABA-gated Cl(-) channel to an open state. PMID- 12559122 TI - Competitive GABA(A) receptor antagonists increase the proportion of functional high-affinity alpha6 subunit-containing receptors in granule cells of adult rat cerebellum. AB - To investigate the properties of alpha6 subunit-containing GABA(A) receptors, whole-cell patch-clamp recordings were made from granule cells in adult rat cerebellar slices. In control, only currents evoked by low concentrations of GABA were significantly reduced in amplitude by furosemide, the alpha6 subunit containing receptor antagonist. However, in the presence of competitive GABA(A) receptor antagonists, the furosemide block of currents evoked by higher GABA concentrations was markedly increased. Zinc, which preferentially blocks alpha6 subunit-containing receptors, also produced an increased block in the presence of bicuculline. To investigate whether similar effects occurred at synaptic receptors, inhibitory postsynaptic currents (IPSCs) were recorded. In most cells, furosemide produced little or no reduction in evoked IPSC amplitude. However in the presence of SR95531, a competitive antagonist, furosemide markedly reduced IPSC amplitude. One hypothesis, which could account for these observations, is that competitive antagonists prevent the continual activation of alpha6beta2/3gamma2 receptors by endogenous GABA and thus prevent their desensitisation. This hypothesis appears feasible as prolonged applications of low concentrations of GABA to recombinant alpha6beta2gamma2s receptors resulted in their desensitisation. PMID- 12559123 TI - Modification of the philanthotoxin-343 polyamine moiety results in different structure-activity profiles at muscle nicotinic ACh, NMDA and AMPA receptors. AB - Voltage-dependent, non-competitive inhibition by philanthotoxin-343 (PhTX-343) analogues, with reduced charge or length, of nicotinic acetylcholine receptors (nAChR) of TE671 cells and ionotropic glutamate receptors (N-methyl-D-aspartate receptors (NMDAR) and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPAR)) expressed in Xenopus oocytes from rat brain RNA was investigated. At nAChR, analogues with single amine-to-methylene or amine-to ether substitutions had similar potencies to PhTX-343 (IC(50)=16.6 microM at -100 mV) whereas PhTX-(12), in which both secondary amino groups of PhTX-343 were replaced by methylenes, was more potent than PhTX-343 (IC(50)=0.93 microM at -100 mV). Truncated analogues of PhTX-343 were less potent. Inhibition by all analogues was voltage-dependent. PhTX-343 (IC(50)=2.01 microM at -80 mV) was the most potent inhibitor of NMDAR. At AMPAR, most analogues were equipotent with PhTX-343 (IC(50)=0.46 microM at -80 mV), apart from PhTX-83, which was more potent (IC(50)=0.032 microM at -80 mV), and PhTX-(12) and 4,9-dioxa-PhTX-(12), which were less potent (IC(50)s>300 microM at -80 mV). These studies show that PhTX-(12) is a selective nAChR inhibitor and PhTX-83 is a selective AMPAR antagonist. PMID- 12559124 TI - Role of AMPA and NMDA receptors in the nucleus accumbens shell in turning behaviour of rats: interaction with dopamine receptors. AB - The role of AMPA and NMDA receptors in the shell of the nucleus accumbens in turning behaviour of rats was investigated. Unilateral injection of the AMPA receptor agonist, AMPA (0.25, 0.4, 0.5 and 1 microg), into the shell of the nucleus accumbens dose-dependently produced contraversive pivoting, namely tight head-to-tail turning marked by abnormal hindlimb backward stepping, while injection of AMPA (0.5 microg) into the core produced only a marginal effect. This shell-specific AMPA effect was dose-dependently inhibited by the AMPA receptor antagonist, NBQX (1 and 10 ng), which alone did not produce turning behaviour. The AMPA-induced pivoting was also dose-dependently inhibited by the non-competitive NMDA receptor antagonist, MK-801 (0.1 and 0.5 microg). Neither MK 801 (0.1, 0.5 and 5 microg) nor the NMDA receptor agonist, NMDA (0.5 and 1 microg), injected unilaterally into the shell, produced turning behaviour. Unilateral injection of a mixture of dopamine D(1) (SKF 38393, 5 microg) and D(2) (quinpirole, 10 microg) receptor agonists into the shell has been found to elicit contraversive pivoting. The dopamine D(1)/D(2) receptor antagonist, cis-(Z) flupentixol (1 and 10 microg), injected into the shell, in doses known to block dopamine D(1)/D(2) receptor-mediated pivoting, also significantly inhibited AMPA (0.5 microg)-induced pivoting. Moreover, both NBQX (1 and 10 ng) and MK-801 (0.1 and 0.5 microg), injected into the shell, significantly inhibited dopamine D(1)/D(2) receptor-mediated pivoting. It is therefore concluded that unilateral stimulation of AMPA receptors in the shell of the nucleus accumbens can elicit contraversive pivoting, and that both AMPA and dopamine D(1)/D(2) receptors play a critical role in shell-specific pivoting in contrast to NMDA receptors that at best play only a modulatory role. PMID- 12559125 TI - The selective iGluR1-4 (AMPA) antagonist LY300168 attenuates morphine-withdrawal induced activation of locus coeruleus neurons and behavioural signs of morphine withdrawal. AB - Previously, we have shown that the AMPA (iGluR1-4) antagonist LY293558 attenuates the morphine-withdrawal-induced activation of locus coeruleus neurons and behavioral signs of morphine withdrawal. However, LY293558 has since been shown to also have affinity for one subtype of kainate receptor (iGluR5). In this study, we examined the effects of a selective antagonist of iGluR1-4 receptors, LY300168 (GYKI 53655), and a selective antagonist of iGluR5 receptors, LY382884, on the morphine-withdrawal-induced activation of locus coeruleus neurons and behavioral signs of morphine withdrawal. In in vivo recordings from anesthetized rats, pretreatment with LY300168 (0.3-3.0 mg/kg, s.c.), but not LY382884 (at a dose known to have central effects; 100 mg/kg, s.c.) attenuated the morphine withdrawal-induced activation of LC neurons. In unanesthetized, morphine dependent rats, pretreatment with LY300168 (0.3-3.0 mg/kg, s.c.), but not LY382884 (100 mg/kg, s.c.), suppressed the severity and occurrence of naltrexone precipitated morphine-withdrawal signs. These results indicate iGluR1-4 (AMPA) receptors, but not iGluR5 receptors, play an important role the morphine withdrawal-induced activation of LC neurons and a subset of behavioral signs of morphine withdrawal. In addition, selective AMPA antagonists may have therapeutic effects in man for the treatment of withdrawal from opiates and other drugs of abuse. PMID- 12559126 TI - Chronic fluoxetine induces opposite changes in G protein coupling at pre and postsynaptic 5-HT1A receptors in rat brain. AB - Chronic treatment with the antidepressant fluoxetine may lead to changes in the properties of pre- and postsynaptic 5-HT(1A) receptors due to modifications in the receptor-G protein coupling process. We have evaluated, in rats, the effect of chronic fluoxetine (10 mg/kg/day) at brain 5-HT(1A) receptors using different techniques. The density of 5-HT(1A) receptors was unchanged in fluoxetine-treated rats vs. vehicle group. Stimulation of [(35)S]GTPgammaS binding induced by (+/-)8 OH-DPAT was significantly attenuated in dorsal raphe nucleus after fluoxetine (+3.7 vs. +31.2% in vehicle). The inhibition of dorsal raphe firing by (+/-)8-OH DPAT (ED(50) in vehicle = 2.1 microg/kg, i.v.) was also attenuated in rats treated with fluoxetine (ED(50)=4.7 microg/kg). In contrast, a significant increase on (+/-)8-OH-DPAT-induced stimulation of [(35)S]GTPgammaS binding was observed in CA(1) (+53.4 vs.+20.2% in vehicle) and dentate gyrus (+105.7 vs. +52.6% in vehicle) but not in entorhinal cortex. Our data demonstrate that fluoxetine-induced desensitization of 5-HT(1A) autoreceptors occurs at G protein level. Moreover, a relevant finding is the region-specific hypersensitivity of postsynaptic 5-HT(1A) receptors, in the hippocampus but not in entorhinal cortex, following chronic fluoxetine. These differential adaptive changes in brain 5 HT(1A) receptors could underlie the mechanism of action of antidepressants and also contribute to their clinical effects. PMID- 12559127 TI - Behavioral sensitization to quinpirole is not associated with increased nucleus accumbens dopamine overflow. AB - This study assessed the relationship between extracellular nucleus accumbens (NAc) dopamine (DA) concentrations and sensitized locomotor activation following repeated administration of the DA D2-like receptor agonist quinpirole. Locomotor activity measures and nucleus accumbens microdialysis samples were collected concurrently in response to the first (acute) and tenth (repeated) quinpirole injection (0.5 mg/kg s.c., every other day). Results indicate that acute quinpirole produced locomotor activation and that repeated quinpirole resulted in locomotor sensitization. Acute quinpirole significantly decreased the detection of extracellular concentrations of DA and the DA metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the NAc. Following repeated quinpirole, basal NAc DA levels were decreased, whereas basal DOPAC levels were increased. Nevertheless, quinpirole challenge elicited a significant decrease in DA, DOPAC and HVA following repeated treatment. In addition, although acute quinpirole did not affect NAc levels of the serotonin metabolite 5-hydroxyindolacetic acid (5-HIAA), quinpirole challenge produced a significant increase in 5-HIAA levels following repeated treatment. Taken together, these data indicate that functional DA autoreceptor subsensitivity is not a necessary condition for the expression of behavioral sensitization to quinpirole. Instead, it appears that behavioral sensitization to quinpirole occurs predominantly as a consequence of neuroadaptations that are post-synaptic to DA release. PMID- 12559128 TI - Mild thyroid hormones deficiency modifies benzodiazepine and mu-opioid receptor binding in rats. AB - The effects of a mild hypothyroidism condition on benzodiazepine (BDZ) and mu opioid receptor levels was investigated. Female Wistar rats were randomly divided into two groups: 1) hypothyroid rats (n=7), which received methimazole (60 mg/kg per day) in drinking water for four weeks, and 2) euthyroid rats (n=8), which drank only tap water. Animals were sacrificed and their brains were used for autoradiography experiments. When compared to the euthyroid group, the hypothyroid group presented reduced benzodiazepine receptor binding in medial amygdala (24%) and high mu-receptor levels in frontal (25%), sensorimotor (65%) and temporal (29%) cortices, basolateral amygdala (50%) and ventroposterior thalamic nucleus (49%). The present data suggest that alterations in BDZ and mu receptor binding could be associated with the higher excitability observed in animals with triiodothyronine (T(3)) deficiency. PMID- 12559129 TI - A tachykinin-like factor increases glutamate toxicity in rat cerebellar granule cells. AB - Tachykinins (TKs), which include substance P, neurokinin A and neurokinin B, constitute a group of neuropeptides widely expressed in the CNS where they play several functions connected with neural modulation often in synergy with glutamate excitatory transmission. The aim of this study was to assess whether TKs modulate glutamate response of in vitro cultured cerebellar granule neurons and whether GSA (glutamate-sensitizing activity), a peptide released by these neurons, belongs to the TKs family. Treatment with substance P and other neurokinin 1 receptor (NK1) agonists does not affect the response of cerebellar granule neurons to glutamate toxicity. On the contrary, agonists neurokinin 2 receptor (NK2) and neurokinin 3 receptor (NK3) agonists increase, in a dose and time dependent fashion, the response of the same neurons to glutamate. MEN 10,627, a selective NK2 receptor antagonist, and (Trp(7),betaAla(8)) NKA (4-10), a selective NK3 receptor antagonist inhibit not only the sensitizing action to glutamate of their respective agonists. These antagonists almost equally reduce the glutamate-sensitizing activity of GSA. Such activity is also abolished in the presence of a polyclonal antibody directed against neurokinin B (NKB). These findings indicate that TKs increase glutamate sensitivity in cerebellar granule neurons and that the GSA previously detected in conditioned media of the same cultured neurons belongs to the TK family although its primary structure as compared to known TKs remains to be established. PMID- 12559130 TI - Lysine 195 and aspartate 196 in the first extracellular loop of the VPAC1 receptor are essential for high affinity binding of agonists but not of antagonists. AB - The role in ligand recognition and receptor activation of two adjacent charged residues (lysine 195 and aspartate 196) in the first extracellular loop of the human VPAC(1) receptor was investigated in stably transfected CHO cells expressing the wild type or point mutated receptors.Replacement of lysine 195 by glutamine or of aspartate 196 by asparagine reduced the agonists' ability to stimulate adenylate cyclase activity; VIP behaved like a partial agonist and a partial agonist behaved as an antagonist. The receptor's capacity to recognize agonists was reduced but antagonists' affinity was unaffected. Both results suggesting that the two charged residues are essential for VPAC(1) receptor activation. On the other hand, the double mutant was less severely affected than single mutants suggesting that hydrogen bonds may partially compensate the loss of charged residues. But the inversion of the residues affected receptor recognition and activation more markedly suggesting that the two charged residues do not interact directly. PMID- 12559131 TI - Modulation of P2X3 receptors by Mg2+ on rat DRG neurons in culture. AB - On nociceptive neurons the commonest response to ATP is a rapidly desensitizing current mediated by P2X(3) receptors and believed to be involved in certain forms of pain. P2X(3) receptor recovery from desensitization is a slow process. We studied whether Mg(2+) might modulate such ATP-evoked currents on rat cultured DRG neurons, and thus account for its analgesic action in vivo. Transient increases in extracellular Mg(2+) strongly and reversibly depressed ATP currents which had not recovered from desensitization. Ca(2+)-free solution had the same action as Mg(2+). High Mg(2+) or Ca(2+)-free modulation depended on exposure length to modified divalent cation solutions, whereas it was independent from membrane potential or intracellular Ca(2+) buffering. Paired-pulse protocols showed that high Mg(2+) or Ca(2+)-free medium delayed ATP receptor recovery from desensitization, while leaving desensitization onset apparently unchanged. Tests with various concentrations of Ca(2+) and Mg(2+) showed that the depressant action by Mg(2+) was primarily due to functional antagonism of a facilitatory effect of Ca(2+) on ATP receptor function. The present results suggest that, on sensory neurons, P2X(3) receptors could be inhibited by high Mg(2+) or lack of Ca(2+), representing a negative feedback process to limit ATP-mediated nociception. PMID- 12559132 TI - Tuberoinfundibular peptide of 39 residues (TIP39): molecular structure and activity for parathyroid hormone 2 receptor. AB - The neuropeptide TIP39 was recently purified from bovine hypothalamus based on the ability of the peptide to activate the parathyroid hormone 2 receptor (PTH2R) ( Nat. Neurosci. 2 (1999) 941). PTH2R is abundantly expressed in the nervous system, and its expression pattern suggests that it may play a role in modulation of pituitary function and in nociception. Towards understanding the physiological role of TIP39 and PTH2R, we cloned human, mouse and rat TIP39 gene. Our results revealed that: (1) the mature peptide is processed from a precursor; (2) TIP39 peptide is highly conserved among species; and (3) TIP39 from all species activates adenylyl cyclase and elevates intracellular calcium levels through PTH2R. We also defined and compared the structure-activity relationship of TIP39 on both activation of adenylyl cyclase and calcium mobilization pathways through PTH2R, finding common and differential determinants of TIP39 that are required for these pathways. Furthermore, we observed that TIP39 elevates intracellular calcium levels in primary dorsal root ganglion neurons whereas the peptide inactive on PTH2R do not, suggesting that TIP39 may activate these neurons important for nociception in vivo through PTH2R-dependent mechanisms. PMID- 12559133 TI - The effects of the shallow and the deep endotracheal suctioning on oxygen saturation and heart rate in high-risk infants. AB - An experimental study involving repeated-measures within subjects was conducted to examine the effects of shallow and deep endotracheal suctioning (ETS) on the SpO(2) and HR in 27 ventilated high-risks infants. The order in which subjects received the ETS protocol was randomly assigned. The results showed no significant changes in both the SpO(2) and HR responses before, during and after ETS between the two ETS protocols. It is concluded that when there is no beneficial effect of performing deep ETS, it should not be carried out due to the potential hazard of direct irritation with more negative pressure on the airways in high-risk infants. PMID- 12559134 TI - The effects of a research utilization in-service program on nurses. AB - The objective of this quasi-experimental design study was to evaluate an 8-week course of research utilization training provided for nurses. Eighty-nine nurses participated in the complete study. Age and education background of the nurses in the control group matched that of the participants in the experimental group. The research instruments included a scale of attitudes toward nursing research, a scale of perceived support for nursing research, a research participation questionnaire, and a research utilization questionnaire. Repeated measures analysis of covariance and the Mann-Whitney U test were adopted for statistical analysis. The results showed that there were significant differences in attitudes between the two groups toward research and perceived support of institutions. Participation in research also differed significantly when analyzed at posttests 2 and 6 months after the course. There was no significant difference in research utilization. These results suggest that continuous consultation and assistance should be provided to the nurses after the course, so as to implement the results of research utilization. PMID- 12559135 TI - Correlation of mental health problems with psychological constructs in adolescence: final results from a 2-year study. AB - In recent years, it has been widely witnessed that a surprising number of adolescents suffer emotional and mental health problems, and such turmoil is very often carried over into adulthood with serious implications for adjustment during the post-adolescent years. On this point, mental health problems in Korea are only now being considered crucial factors in the health status of adolescents and important public and social issues. It is also true that studies concerning the link between adolescents' mental health problems and their psychology are limited. The purposes of this study were to investigate mental health problems of Korean adolescents, to reveal factors affecting their negative mental health and to explore a possible relationship between mental health problems and psychological variables. 2052 Korean adolescents selected randomly from junior high and high schools in Seoul, Korea were surveyed. Korean Symptom Checklist, Health Locus of Control Scale, Self-efficacy Scale and Self-esteem Scale were used to identify mental health problems and psychological variables of adolescents. Results indicated that Korean adolescents showed high prevalence in interpersonal sensitivity, depression, anxiety and hostility. In addition, the findings revealed that there were significant differences in adolescents' mental health problems between gender and age. Furthermore, results revealed that the adolescents' mental health problems were statistically correlated with psychological variables. This study provides significant information for the relatively unstudied Korean adolescents and also has the potential to influence the development of better mental health programs for adolescents. PMID- 12559136 TI - Student nurses' attitudes to pain relieving drugs. AB - This paper reports the results of a longitudinal survey of 217 student nurses undertaken during their common foundation programme (CFP), which comprises the first 18 months of the course. The study explored changes during the CFP in students' perceptions of the risk of addiction and their attitudes towards analgesics. The results revealed that although the students had a more accurate view of the risk of addiction by the end of the CFP there was still an exaggerated fear of the risk of addiction in over half the students. Follow-up interviews with 14 students also indicated that students displayed a more general wariness in relation to the use of analgesics. PMID- 12559137 TI - Perceptions of nursing: a study involving nurses, nursing students, patients and non-nursing students. AB - This paper describes a study of perceptions of nursing by nursing students, qualified nurses (United Kingdom and Spain), patients and non-nursing students. A survey method was used with a longitudinal panel element incorporating a 35-item version of the Nursing Dimensions Inventory translated into Spanish. Data were analysed using principal component analysis, congruence analysis, Mokken scaling, correlation, ANOVA, t-test and Cronbach's alpha. Patients may perceive nursing differently from nursing students, nurses (in the UK and Spain) and non-nurses. These findings are discussed in the context of nurse education and patient care. PMID- 12559138 TI - Psychiatric nurses' views on criteria for psychiatric intensive care: acute and intensive care staff compared. AB - AIM: To explore and investigate differences between the views of qualified nurses working in psychiatric intensive care units (PICUs) and acute care wards on which patients are appropriate for PICU care. BACKGROUND: Previous research on the area of psychiatric intensive care highlights the great differences that exist in all aspects of service provision, from unit size and staffing levels to treatment approaches and physical environment. One of the most common areas of controversy is the type of client behaviour that warrants admission to the PICU. METHOD: Structured interviews of 100 qualified nursing staff (in the London area, England) working on either acute or PICU wards were used to gather data on appropriate and inappropriate referral to PICUs. Comments made during the course of the interviews were also collected and subjected to content analysis. FINDINGS: There was evidence to support the hypothesis that acute ward staff considered patients suitable for PICU care at a lower level of risk than PICU staff thought appropriate. In comparison to acute ward nurses, those working in PICUs attended to a broader range of factors when considering suitability for admission to PICU. Appropriate reasons for transfer fell into five groups: risk to others; risk of intentional harm to self; risk of unintentional harm to self; therapeutic benefit from the PICU environment; and legitimate acute ward care problem. Inappropriate reasons for transfer fell into four groups: low risk to others and/or self; illegitimate acute admission care problems; patient belongs elsewhere; policy issues. CONCLUSION: The study opens up a range of issues not previously studied in relation to the use of PICUs and the intricate relationship of this use with the available acute care wards and other services. These findings and their implications for the care of acute and disturbed psychiatric patients are discussed. PMID- 12559139 TI - A salutogenetic perspective on how men cope with major depression in daily life, with the help of professional and lay support. AB - Worldwide, major depression is one of the main causes of premature death and lowered functional capacity, and its importance will increase in the coming years. At the same time, there is a gap in service provision, and the needs of depressed patients are often not met. Therefore, the aim of this study was to describe how men (N=12) with major depression (DSM-IV) cope with daily life with the help of professional and lay support. The men were selected by strategic sampling and analysed with an approach inspired by phenomenography. Four descriptive categories emerged: being unburdened, restoring one's health, feeling involved, and finding a meaning. A man receives help and support from health care in restoring his health but his own path to getting well is to resume his place in the public domain. To help him do so, lay support is vital, and an important task for the nurse is, therefore, to support him in maintaining his existing social network or in building a new one. PMID- 12559140 TI - The effect of medication management training on community mental health nurse's clinical skills. AB - Background. There is evidence that compliance therapy improves treatment adherence and clinical outcomes in patients with schizophrenia. Community Mental Health Nurses (CMHNs) are ideally placed to deliver compliance therapy but require training to develop the necessary clinical skills.Aim. To explore whether a brief medication management training package is effective in developing the compliance therapy skills of CMHNs.Method. The study had a within subjects repeated measures design. A representative sample of 52 CMHNs entered the study. They performed a role-play task pre- and post-training that was videotaped and blind rated by an independent rater using the Cognitive Therapy Scale (CTS). Knowledge was assessed pre- and post-training using a knowledge about medication management questionnaire. Trainees also completed a satisfaction with training questionnaire at the end of the course.Results. Following training there was a statistically significant improvement in mean scores on the primary measure of skills, the CTS (mean pre-training CTS-total score 13.88, mean post-training CTS total score 31.12; p<0.01). There was also a significant categorical improvement in the number of trainees who demonstrated satisfactory skills. Knowledge about medication management was significantly improved and trainees reported that training was acceptable and relevant to their clinical practice.Conclusions. In this uncontrolled study training improved the medication management skills of CMHNs. PMID- 12559141 TI - Not knowing--the problem of pain in children with severe neurological impairment. AB - This study uses a grounded theory approach to explore the diagnostic and clinical decision-making processes used by parents and healthcare professionals in relation to pain in children with severe to profound neurological impairment. Three forms of knowledge are required for optimal pain assessment and management (1) knowing the child, (2) familiarity with children with the same or similar conditions and (3) knowing the science. Pain relief can be compromised by systems of care that fragment rather than integrate care. A model which integrates knowledge of child, population and science is proposed together with an intersubjective attitude to assessing pain. PMID- 12559142 TI - Coping and adaptation at six months after stroke: experiences with eating disabilities. AB - Stroke produces a range of enduring impairments and survivors' coping and adaptation styles are influential features of life after stroke. Many stroke sequelae affect ability to eat but survivors' perceptions and responses to these have not been explored.Methods. Survivors of a cohort of patients admitted to hospital with acute stroke March 1998-April 1999 participated in semi-structured interviews in their homes at 6 months post-stroke. Interviews were tape recorded and transcribed; 113 interviews with eating-disabled subjects were entered onto QSR NUD*IST 4 for thematic analysis.Findings. Two major emergent themes of 'getting back to normal' and 'getting by' were revealed which encompassed a range of responses in relation to food and eating. A high level of congruence was demonstrated with pre-existent frameworks but with some unique features. Reportage demonstrated non-linear and inconsistent effects of impairments within patients' lives and the importance of this topic for survivors and healthcare professionals was clear. PMID- 12559143 TI - Spirituality or psychosis?--An exploration of the criteria that nurses use to evaluate spiritual-type experiences reported by patients. AB - Spiritual experiences and psychotic symptoms have many aspects of form and content in common. Despite this, clinicians make judgements about the pathology of these experiences and base care-plans on these judgements. Semi-structured interviews incorporating vignettes of spiritual-type experiences were given to 14 UK mental health nurses. This revealed that the nurses employed a complex and inter-relating set of criteria when evaluating spiritual-type experiences. The nature of the experience was considered, but the outcome of the experience (positive or negative) was an important evaluative factor, together with the personal and cultural context in which the experience occurred. The nurses demonstrated a tolerance of ambiguity and the need for awareness of their own subjectivity. They emphasised the importance of close-engagement with patients to achieve a rounded and holistic view of the patient's experience. They also emphasised the importance of team working in reducing idiosyncratic decision making. PMID- 12559144 TI - A controlled trial of psycho-educational interventions in preparing Chinese women for elective hysterectomy. AB - The objective of this study was to investigate the effect of a cognitive intervention (distraction and reappraisal) with information given pre-operatively on post-operative outcomes of Chinese women having an abdominal hysterectomy. Using a controlled trial, 48 Chinese women having elective hysterectomy received the cognitive intervention with information (experimental group). A control group (n=48) received information alone. Outcome measures used in the study were post operative anxiety and pain, requests for analgesia and patient satisfaction. The results show that women in the experimental group reported lower post-operative anxiety scores, lower pain scores and higher levels of satisfaction than women in the control group. There were no statistically significant differences in post operative requests for analgesia between the groups. It is concluded that a cognitive intervention such as distraction and reappraisal may have significant clinical benefits and improve the care of women having an elective hysterectomy. PMID- 12559146 TI - Theory of mind and psychopathy: can psychopathic individuals read the 'language of the eyes'? AB - There have been suggestions that Theory of Mind (ToM) impairment might lead to aggressive behaviour and psychopathy. Psychopathic and matched non-psychopathic individuals, as defined by the Hare Psychopathy Checklist [The Hare Psychopath Checklist-Revised, 1991] completed the 'Reading the Mind in the Eyes' ToM Test [Journal of Child Psychology and Psychiatry, 1997;38:813]. This test requires the self-paced identification of mental states from photographs of the eye region alone. Results indicated that the psychopathic individuals did not present with any generalised impairment in ToM. The data are discussed with reference to the putative neural system mediating performance on this task and models of psychopathy. PMID- 12559147 TI - Impairment of recognition of disgust in Chinese with Huntington's or Wilson's disease. AB - The selective involvement of the basal ganglia in recognition of the facial expression of disgust was investigated by examining a group of six symptomatic Huntington's disease patients and 32 Wilson's disease patients in China. Morphed photographs of facial expressions covering happiness-surprise-fear-sadness disgust-anger were used and the patients were asked to label each photo. Other measures assessed basic cognitive functions and perception of non-emotion facial information, such as perception of gender, age, gaze direction, and recognition of unfamiliar as well as famous people. There was dissociation between the perception of emotions and other facial information, and between impairment of recognition of disgust and other emotions. The basal ganglia are the overlapping substrate involved in both Huntington's and Wilson's disease, although each has its own other lesions. The differentially severe impairment of recognition of disgust in the Chinese Huntington's disease and Wilson's disease patients strengthens the view that basal ganglia are selectively involved in processing the emotion of disgust. PMID- 12559148 TI - Inappropriate association of semantics and context to novel stimuli can give rise to the false recognition of unfamiliar people. AB - This paper reports further experiments with a patient (MR) who has a tendency to claim that unfamous people are familiar together with good ability at identifying truly famous people. The first experiment examines the role that the typicality of stimuli plays in his false recognition. Although, typicality may have some influence over false recognition (as it does for normal controls) there is little evidence to suggest that MR is over-reliant on such information. It is unlikely that perceptual fluency can entirely explain his deficit. This is bolstered by a further study using morphed images of famous and unfamous faces, suggesting that false recognition is associated with inappropriate retrieval of semantic biographical information. It is argued that MR's judgement of 'fame' is subjectively appropriate, given the information that he retrieves. This information is thought to derive from currently activated, or recently activated, contextual information which becomes inappropriately bound to the novel stimulus, giving rise to a false sense of familiarity. These findings underscore the importance of viewing memory as an attributional process, whereby current mental constructions/processes are attributed to some event(s) in the past. PMID- 12559149 TI - Reality monitoring and visual hallucinations in Parkinson's disease. AB - Between 8 and 40% of Parkinson disease (PD) patients will have visual hallucinations (VHs) during the course of their illness. Although cognitive impairment has been identified as a risk factor for hallucinations, more specific neuropsychological deficits underlying such phenomena have not been established. Research in psychopathology has converged to suggest that hallucinations are associated with confusion between internal representations of events and real events (i.e. impaired-source monitoring). We evaluated three groups: 17 Parkinson's patients with visual hallucinations, 20 Parkinson's patients without hallucinations and 20 age-matched controls, using tests of visual imagery, visual perception and memory, including tests of source monitoring and recollective experience. The study revealed that Parkinson's patients with hallucinations appear to have intact visual imagery processes and spatial perception. However, there were impairments in object perception and recognition memory, and poor recollection of the encoding episode in comparison to both non-hallucinating Parkinson's patients and healthy controls. Errors were especially likely to occur when encoding and retrieval cues were in different modalities. The findings raise the possibility that visual hallucinations in Parkinson's patients could stem from a combination of faulty perceptual processing of environmental stimuli, and less detailed recollection of experience combined with intact image generation. PMID- 12559150 TI - Cognitive estimation in patients with probable Alzheimer's disease and alcoholic Korsakoff patients. AB - Cognitive estimation is an important function in daily living. In early studies it was proposed that estimation deficits are associated with frontal lobe damage and executive dysfunctions. In this study, we assessed Alzheimer patients and patients with alcoholic Korsakoff's syndrome with a newly developed cognitive estimation task. We compared their performance with respect to different dimensions of estimation ('size', 'weight', 'quantity', and 'time') and to various error types. Compared to healthy controls, both patient groups were strongly impaired in all tested estimation dimensions, with Alzheimer patients performing generally worse than Korsakoff patients, except for the dimension "time". Alzheimer as well as Korsakoff patients produced so-called 'bizarre errors' and errors in the choice of the correct unit of measurement. In both patient groups cognitive estimation correlated highly with general knowledge. The production of bizarre errors and unit errors correlated with general knowledge as well as with working memory and executive functions. Results support the main assumptions of a model of cognitive estimation, described in the discussion, that specific parts of the semantic memory system as well as executive functions, in form of a plausibility check of the generated answer, are involved in the process of cognitive estimation. PMID- 12559151 TI - An fMRI study investigating cognitive modulation of brain regions associated with emotional processing of visual stimuli. AB - Brain regions modulated by cognitive tasks during emotional processing were investigated using fMRI. Participants performed indirect and direct emotional processing tasks on positive and negative faces and pictures. We used a multivariate technique, partial least squares (PLS) to determine spatially distributed patterns of brain activity associated with different tasks and stimulus conditions, as well as the interaction between the two. The pattern of brain activity accounting for the most task-related covariance represented a task x stimulus interaction and distinguished indirect processing of pictures and direct processing of faces from direct processing of pictures and indirect processing of faces. The latter two conditions were characterised by limbic (e.g. amygdala, insula, thalamus) and temporal lobe activity, in addition to greater activity in the ventral prefrontal cortex. Indirect and direct processing of pictures and faces, respectively, were represented by more dorsal prefrontal and parietal activity. These findings indicate that brain activity during processing of emotional content is critically dependent on both the type of stimulus and processing task. In addition, these results support the idea that the pattern of activity in the emotional network can be influenced in a 'top-down' fashion via cognitive factors such as attentional control, and as such, have important clinical implications for emotional disorders, such as depression and anxiety. PMID- 12559152 TI - Interhemispheric visual interaction in a patient with posterior callosectomy. AB - The role of anterior commissure (AC) and anterior parts of corpus callosum in visual interactions was investigated in a partial split-brain patient whose posterior and middle parts of the corpus callosum were resected surgically leaving intact only a thin portion of anterior corpus callosum. Although the primary visual areas of the two hemispheres are disconnected in the patient, we found that visual distracters presented to one hemisphere (in a crowding paradigm) impaired recognition of the target stimulus presented to the other hemisphere. The normal control group showed the same result. To rule out the possible contribution of subcortical areas to this interaction, we repeated the same crowding task with texture-defined stimuli. The patient again showed an interhemispheric interaction, even though subcortical structures respond poorly or do not respond at all to texture defined shapes. Despite the evidence for interhemispheric interaction, a classic match-to-sample task confirmed that the patient was unable to explicitly report when stimuli in left and right hemifields were the same or different. Similarly, in a search task, the patient's reaction time was unaffected by distracters in the hemifield opposite the target whereas normals' response time was affected. Considering the dissociation between these two tasks, we conclude that the anterior commissure and/or the anterior corpus callosum contribute to interhemispheric interactions in the attentional selection of location. PMID- 12559153 TI - Perception and production of facial and prosodic emotions by chronic CVA patients. AB - Variable etiology, limited testing of emotions and inclusion of patients in acute and chronic phases have made it difficult to specify the extent of right hemisphere involvement in the processing of emotional material. In addition, there is an absence of data concerning CVA patients' long-term abilities to process emotional information. Two groups of subjects with unilateral brain damage (15 RBD, 17 LBD), matched for chronicity (minimum 12 months post-CVA), etiology (ischemic CVA), duration of hospitalization and other variables, and an appropriate control group participated in two experiments to address these concerns. In the first experiment, subjects were given a series of tasks (discrimination, identification, imitation, production on request) to assess their processing of facial expressions of the six universal emotions [P. Ekman, W. Friesen, Unmasking the face, Prentice-Hall, Englewood Cliffs, 1975]. The results showed that three emotions (surprise, happiness, fear) discriminate between RBD and LBD patients and controls, with RBD subjects performing worse than the other groups on the identification task only. Using tasks of the same nature, the second experiment investigated the processing of emotional prosody. The results showed that three emotions (fear, sadness, anger) discriminate between RBD and LBD patients and controls, with RBD subjects again performing worse than the others on the discrimination, imitation and production on request tasks. LBD subjects performed as well as normal controls on almost all tasks. The RBD subjects were the only ones who showed relatively consistent impairment in the processing of both facial and prosodic emotional information. Taken together, these data strongly suggest that the right hemisphere is preferentially involved in processing emotional information in the chronic phase of brain damage. PMID- 12559154 TI - Ideomotor limb apraxia in Huntington's disease: implications for corticostriate involvement. AB - Ideomotor limb apraxia, a disorder of goal-directed movement, has been attributed to lesions in the frontal and parietal lobes, but the role of subcortical structures is less certain. In order to determine its prevalence in a disorder affecting the basal ganglia and corticostriatal connections, we examined imitation of hand gestures in Huntington's disease (HD) patients. We also assessed the relationship between apraxia and cognitive and motor dysfunction in an effort to better understand the neural underpinnings of apraxia in HD. If damage restricted to the basal ganglia produces ideomotor limb apraxia, then we would expect to find evidence of apraxia in patients who were early in the disease course when selective striatal damage is most common. Such a pattern, however, was not found in our sample. Instead, patients with greater neurological impairment and with a longer duration of disease were more likely than less affected patients to demonstrate apraxia. Apraxia was not related to severity of chorea, but was associated with greater impairment in eye movements, voluntary movements, and verbal fluency. These findings suggest that apraxia in HD results from damage to the corticostriate pathways and the basal ganglia rather than from damage restricted to the basal ganglia. PMID- 12559156 TI - Temporary and permanent signs of interhemispheric disconnection after traumatic brain injury. AB - The corpus callosum is frequently damaged by closed head traumas, and the resulting deficits of interhemispheric communication may vary according to the specific position of the lesion within the corpus callosum. This paper describes a single case who suffered a severe traumatic brain injury resulting in a lesion of the posterior body of the corpus callosum. Among the classical symptoms of interhemispheric disconnection, left hand anomia, left upper limb ideomotor dyspraxia, left visual field dyslexia and dysnomia, and left ear suppression in a dichotic listening task were observed shortly after the injury but recovered completely or almost completely with the passage of time. The only symptom of interhemispheric disconnection which was found to persist more than 4 years after the injury was an abnormal prolongation of the crossed-uncrossed difference in a simple visuomotor reaction time task. This prolongation was comparable with that observed in subjects with complete callosal lesions or agenesis. The results suggest that the posterior body of the corpus callosum may be an obligatory interhemispheric communication channel for mediating fast visuo-motor responses. The transient nature of other symptoms of interhemispheric disconnection suggests a relatively wide dispersion of fibers with different functions through the callosal body, such that parts of them can survive a restricted lesion and allow functional recovery of hemispheric interactions. An assessment of the evolution in time of symptoms of interhemispheric disconnection following restricted callosal lesions may reveal fine and coarse features of the anatomo-functional topography of the corpus callosum. PMID- 12559155 TI - Vision for spatial perception and vision for action: a dissociation between the left-right and near-far dimensions. AB - Neuropsychological and psychophysical studies have suggested that two distinct visual sub-systems are responsible for perception and action. One of the main psychophysical arguments for this is based on visual illusion such as the Induced Roelofs Effect (IRE), where the location of a visual target presented with an off centre frame is misperceived when evaluated verbally, but not with a reaching response. This dissociated effect suggests the existence of two independent representations of visual space devoted, respectively, to categorisation and to egocentric localisation of reachable objects. These "cognitive" and "sensorimotor" representations have been assumed to be produced through specific anatomical pathways stemming from the primary visual cortex (respectively, the ventral and dorsal streams). To account for the dissociation found with the IRE, it has been suggested that only the cognitive system is sensitive to contextual information. However this view has been challenged by recent psychophysical studies demonstrating the influence of environmental cues on distance perception and the guiding of movement. In the present study, the IRE is re-evaluated but the near-far and right-left dimensions were dissociated. In agreement with previous findings, our results showed that the IRE in the right-left dimension gives rise to a perceptual misperception of target position with no effect on motor performance. Conversely, when the IRE was induced in the near-far dimension a misperception of the target position affected both perceptual and motor responses. This dissociation indicates that the spatial constraints of the task, and not only the nature of the response, interfere with sensitivity to contextual information leading to visual illusions. It is thus likely that the action system (imputed to the dorsal stream) can be sensitive to contextual information, at least when depth processing is emphasised. PMID- 12559157 TI - Motor cortex hand area and speech: implications for the development of language. AB - Recently a growing body of evidence has suggested that a functional link exists between the hand motor area of the language dominant hemisphere and the regions subserving language processing. We examined the excitability of the hand motor area and the leg motor area during reading aloud and during non-verbal oral movements using transcranial magnetic stimulation (TMS). During reading aloud, but not before or afterwards, excitability was increased in the hand motor area of the dominant hemisphere. This reading effect was found to be independent of the duration of speech. No such effect could be found in the contralateral hemisphere. The excitability of the leg area of the motor cortex remained unchanged during reading aloud. The excitability during non-verbal oral movements was slightly increased in both hemispheres. Our results are consistent with previous findings and may indicate a specific functional connection between the hand motor area and the cortical language network. PMID- 12559158 TI - Interhemispheric cooperation for face recognition but not for affective facial expressions. AB - Interhemispheric cooperation can be indicated by enhanced performance when stimuli are presented to both visual fields relative to one visual field alone. This "bilateral gain" is seen for words but not pseudowords in lexical decision tasks, and has been attributed to the operation of interhemispheric cell assemblies that exist only for meaningful words with acquired cortical representations. Recently, a bilateral gain has been reported for famous but not unfamiliar faces in a face recognition task [Neuropsychologia 40 (2002) 1841]. In Experiment 1 of the present paper, participants performed familiarity decisions for faces that were presented to the left (LVF), the right (RVF), or to both visual fields (BVF). An advantage for BVF relative to both LVF and RVF stimuli was seen in reaction times (RTs) to famous faces, but this bilateral advantage was absent for unfamiliar faces. In Experiment 2, participants classified the expression (happy or neutral) of unfamiliar faces. No bilateral advantage was seen for expressions, although a right hemisphere superiority was seen in terms of higher accuracy for LVF and BVF trials relative to the RVF. Recognition of famous faces (but not of facial expressions) require access to acquired memory representations that may be instantiated via cortical cell assemblies, and it is suggested that interhemispheric cooperation depends on these acquired cortical representations. PMID- 12559159 TI - Binocular vision and prehension in middle childhood. AB - Binocular cues have been shown previously to make an important contribution to the control of natural prehensile movements in adults [Visual Cognition 4 (1997) 113, Vision Research 32 (1992) 1513, Neuropsychologia 38 (2000) 1473]. The present study examined the role of binocular vision in the control of prehension in middle childhood. Fourteen children aged 5-6 years, and 16 children aged 10-11 years reached out and grasped different sized objects at different distances, in either binocular or monocular viewing conditions. In contrast to adult data, many of the principal kinematic indices of the children's reaches were unaffected by the removal of binocular information. The older children, like adults, spent an increased amount of time in the final approach to the object when only monocular information was available. However, both peak wrist velocities and peak grip apertures were unaffected by the removal of binocular information and continued to scale with object properties in the normal way. These results suggest that the use of binocular cues to control prehensile movements is not yet mature at the age of 10-11 years. PMID- 12559160 TI - Less impairment in face imagery than face perception in early prosopagnosia. AB - There have been a number of reports of preserved face imagery in prosopagnosia. We put this issue to experimental test by comparing the performance of MJH, a 34 year-old prosopagnosic since the age of 5, to controls on tasks where the participants had to judge faces of current celebrities, either in terms of overall similarity (Of Bette Midler, Hillary Clinton, and Diane Sawyer, whose face looks least like the other two?) or on individual features (Is Ronald Reagan's nose pointy?). For each task, a performance measure reflecting the degree of agreement of each participant with the average of the others (not including MJH) was calculated. On the imagery versions of these tasks, MJH was within the lower range of the controls for the agreement measure (though significantly below the mean of the controls). When the same tasks were performed from pictures, agreement among the controls markedly increased whereas MJH's performance was virtually unaffected, placing him well below the range of the controls. This pattern was also apparent with a test of facial features of emotion (Are the eyes wrinkled when someone is surprised?). On three non-face imagery tasks assessing color (What color is a football?), relative lengths of animal's tails (Is a bear's tail long in proportion to its body?), and mental size comparisons (What is bigger, a camel or a zebra?), MJH was within or close to the lower end of the normal range. As most of the celebrities became famous after the onset of MJH's prosopagnosia, our confirmation of the reports of less impaired face imagery in some prosopagnosics cannot be attributed to pre-lesion storage. We speculate that face recognition, in contrast to object recognition, relies more heavily on a representation that describes the initial spatial filter values so the metrics of the facial surface can be specified. If prosopagnosia is regarded as a form of simultanagnosia in which some of these filter values cannot be registered on any one encounter with a face, then multiple opportunities for repeated storage may partially compensate for the degraded representation on that single encounter. Imagery may allow access to this more complete representation. PMID- 12559161 TI - Disruptive influences of a cued voluntary shift on coordinated movement in Parkinson's disease. AB - A temporary and/or involuntary stoppage of movement is identifiable in the execution phase of writing, walking, and turning movements in individuals with Parkinson's disease (PD) and may be referred to as freezing. However, the unpredictability of such akinetic impairments has made it difficult to study experimentally. The present study compared PD and age-matched control groups in their ability to coordinate continuous and simultaneous upper limb movements in trials involving two parts. In the first part of each trial, participants performed either in-phase movements (symmetric, simultaneous movement toward and away from the midline of the body), or anti-phase movements (isodirectional). At the midpoint of the trial, they were signaled by an auditory metronome to execute an intentional and voluntarily switch from the coordination currently being performed to the opposite coordination pattern. In the second half of the trial participants were required to maintain performance in the other coordination mode. All trials were paced by an auditory metronome at one of three different speeds (0.75, 1.25, 1.75 Hz). Measures of temporal coordination (relative phase) indicated that overall, participants with PD required significantly longer periods of time to achieve a switch between coordination patterns compared to healthy controls, and experienced greater difficulty changing from the in-phase to anti-phase mode of coordination. As well, movement stoppage was observed in 53.9% of the in-phase to anti-phase switch trials, but in only 15.5% of the anti phase to in-phase trials. We conclude that interruptions to movement execution are most common when switching to coordinated movements that impose greater motor demands in individuals with PD. PMID- 12559162 TI - Visual object and face processing in mild-to-moderate Alzheimer's disease: from segmentation to imagination. AB - Little is known about the fate of higher level visual perception and visual mental imagery in the early stages of Alzheimer's disease (AD). In this study, we assessed these abilities in a group of mild-to-moderate AD patients using tasks selected to satisfy two main criteria. First, they have been shown to be sensitive to impairments of perception and imagery caused by other neurological conditions. Second, they test specific stages of visual perception and cognition in a reasonably selective manner. These stages were (in their normal order of occurrence during perception): the segmentation of different local points of the visual field into regions belonging to distinct objects; the representation of the shapes of these segmented regions in the image; the construction of more abstract shape representations that possess constancy over changes in size, location, orientation or illumination (assessed separately for faces and objects); the use of these perceived shape representations to access stored shape representations; and the access of lexical semantic representations from these high-level visual representations. Additional tasks tested the top-down activation of earlier visual representations from the semantic level in visual mental imagery. Our findings indicate small, but in most cases reliable, impairments in visual perception, which are independent of degree of cognitive decline. Deficits in basic shape processing influenced performance on some higher level visual tasks, but did not contribute to poor performance on face processing, or to the profound deficit on object naming. The latter of these is related to semantic-lexical impairment. PMID- 12559163 TI - Cerebral asymmetry in twins: predictions of the right shift theory. AB - A study of the heritability of lobar brain volumes in twins has introduced a new approach to questions about the genetics of cerebral asymmetry. In addition to the classic comparison between monozygotic (MZ) and dizygotic (DZ) twins, a contrast was made between pairs of two right-handers (RR pairs) and pairs including one or more non-right-hander (non-RR pairs), in the light of the right shift (RS) theory of handedness. This paper explains the predictions of the RS model for pair concordance for genotype, cerebral asymmetry and handedness in healthy MZ and DZ twins. It shows how predictions for cerebral asymmetry vary between RR and non-RR pairs over a range of incidences of left-handedness. Although MZ twins are always concordant for genotype and DZ twins may be discordant, differences for handedness and cerebral asymmetry are expected to be small, consistent with the scarcity of significant effects in the literature. Marked differences between RR and non-RR pairs are predicted at all levels of incidence, the differences slightly larger in MZ than DZ pairs. PMID- 12559164 TI - Sentence priming effects in the two cerebral hemispheres: influences of lexical relatedness, word order, and sentence anomaly. AB - The present study examined lexical, syntactic and semantic message-level contributions to sentence priming in the left (LH) and right (RH) hemispheres. Participants made lexical decisions to laterally presented target words preceded by congruent, incongruent, syntactic, random and neutral incomplete sentences that either contained or did not contain a lexical associate of the target. In the LH, congruent associated sentences facilitated, and incongruent associated sentences inhibited, target word recognition extensively. A similar pattern, but attenuated, was found for the RH. In both the LH and RH, random associated sentences benefited word target recognition very moderately. Syntactic associated sentence primes that contain word-level information embedded in a meaningless message provided the condition in which the RH was benefited but the LH was not. The asymmetry of sentence type context effects in the LH and RH suggests some important differences in message-level processes available to each hemisphere. PMID- 12559165 TI - Effects of prismatic adaptation on judgements of spatial extent in peripersonal and extrapersonal space. AB - Recent research has shown that visuomotor adaptation to a lateral displacement of the visual field induces significant perceptual aftereffects in normal observers, and in right hemisphere patients with spatial neglect [Neuroreport 11 (2000) 1899; Nature 395 (1998) 166]. These findings suggest that adaptive realignment following prism exposure induces a bias in visual space perception, even in tasks that require no visually guided motor response. Given recent neurophysiological and behavioural data suggesting independent visual representations for peripersonal and extrapersonal space, here we asked whether adaptive aftereffects extend beyond participants' immediate reaching space to stimuli presented beyond arms' reach (i.e. in extrapersonal space). Thirty-two participants underwent adaptive realignment to 10 degrees left- or right-displacing wedge-prisms. Before and after adaptation participants performed a visual landmark task that required estimation of the midpoint of horizontal line stimuli. There was a significant rightward shift in visual midpoint judgements following adaptation to left deviating prisms, which was evident in both peripersonal and extrapersonal space. Paradoxically, a significant rightward shift also occurred following adaptation to right-deviating prisms, but only in extrapersonal space. We suggest that the pattern of adaptive aftereffects observed reflects the different reference frames used by participants to perform spatial judgements in peripersonal and extrapersonal space. We also propose that an underlying hemispheric asymmetry in the processing of spatial errors during adaptation may contribute to the direction of aftereffects in both normal observers, and in patients with unilateral lesions. PMID- 12559166 TI - On the hemispheric specialization for categorical and coordinate spatial relations: a review of the current evidence. AB - This article reviews current evidence on the hemispheric specialization hypothesis for two types of spatial relations representations; categorical versus coordinate [Psychol. Rev. 94 (1987) 148; J. Exp. Psychol.: Percept. Perform. 15 (1989) 723]. Categorical representations capture general properties of the spatial structure of a visual stimulus, without defining the exact metric properties. Coordinate representations specify precise spatial locations of objects or parts in terms of metric units. It is claimed that a hemispheric difference in contribution to the computation of both types of spatial relations representations exists, in which the left hemisphere is specialized for the computation of categorical spatial representations while the right hemisphere is specialized for the computation of coordinate ones. Several forms of research (experimental, computer simulations, patient studies and neuroimaging studies) are reviewed. In general, there is convergent evidence for a conceptual separation of coordinate and categorical processing, with strongest indications for a relative right hemisphere advantage in encoding coordinate spatial relations, and weaker support for left hemispheric categorical specialization. The pattern appears to be critically linked to receptive field properties of the two hemispheres and as such is modulated by certain elementary visual characteristics of the displayed stimuli. PMID- 12559167 TI - The amygdala: is it an essential component of the neural network for social cognition? AB - Observations from human subjects with focal brain lesions and animal subjects with experimental lesions have implicated a variety of brain regions in the mediation of social behavior. Previous studies carried out in the macaque monkey found that lesions of the amygdala not only decrease emotional reactivity but also disrupt normal social interactions. We have re-investigated the relationship between amygdala lesions and social behavior in cohorts of mature and neonatal rhesus monkeys who were prepared with selective and complete bilateral ibotenic acid lesions of the amygdaloid complex. These animals display clear alterations in emotional and social behavior. We interpret these changes as due to a loss of the ability to evaluate environmental stimuli as potential threats. However, adult animals with bilateral lesions of the amygdala demonstrate near normal, and even increased, social interactions with conspecifics. Moreover, neonatal animals, prepared with amygdala lesions at 2 weeks of age, also demonstrate species typical social behaviors such as the generation of facial expressions, grooming and play behavior. These results argue against the idea that the amygdala is essential for the interpretation of social communication or for the expression of social behavior. Because it does appear to participate in the evaluation of the "safety" of social interactions, we believe that it does have a role in modulating the amount of social behavior in which an organism will participate. However, our current answer to the question posed in the title of this paper is no! PMID- 12559168 TI - Initiation of angiogenic Kaposi's sarcoma lesions. AB - A new mouse model reported in this issue of Cancer Cell implies that VEGF/KDR mediated paracrine effects induced by the lytic cycle vGPCR signaling protein encoded by human Kaposi's sarcoma-associated herpesvirus (KSHV or HHV8) may be involved in promoting the proliferation of the KSHV latently infected spindle endothelial cells of Kaposi's sarcoma. PMID- 12559169 TI - Take care of your chromosomes lest cancer take care of you. AB - The analysis of compound mouse mutants for nonhomologous end-joining DNA double strand break repair and those deficient for the p53 checkpoint pathway has provided a fascinating look at the carcinogenic consequences of the failure to properly repair DNA damage and to elicit appropriate checkpoints. PMID- 12559170 TI - Human papillomavirus, it's genes...and cancer vaccines. AB - Genital human papillomavirus (HPV) infection is causally linked to the development of cervical cancer, a leading cause of cancer-related mortality in women worldwide. Recent studies demonstrate the effectiveness of virus-like particle-based vaccines to induce neutralizing antibodies against HPV and prevent cervical intraepithelial neoplasia. PMID- 12559171 TI - Proteomics in translational cancer research: toward an integrated approach. AB - Proteomics provides powerful tools for the study of clinically relevant samples in the context of translational cancer research. Here we briefly review applications of gel-based proteomics for the study of bladder and lung cancer using fresh tissue biopsies. In general, these studies have emphasized the potential of the technology for biomarker discovery, as well as for addressing the issue of cancer heterogeneity. PMID- 12559172 TI - Apoptosis-targeted therapies for cancer. PMID- 12559173 TI - Endothelial infection with KSHV genes in vivo reveals that vGPCR initiates Kaposi's sarcomagenesis and can promote the tumorigenic potential of viral latent genes. AB - The Kaposi's sarcoma herpesvirus (KSHV) has been identified as the etiologic agent of Kaposi's sarcoma (KS), but initial events leading to KS development remain unclear. Characterization of the KSHV genome reveals the presence of numerous potential oncogenes. To address their contribution to the initiation of the endothelial cell-derived KS tumor, we developed a novel transgenic mouse that enabled endothelial cell-specific infection in vivo using virus expressing candidate KSHV oncogenes. Here we show that transduction of one gene, vGPCR, was sufficient to induce angioproliferative tumors that strikingly resembled human KS. Endothelial cells expressing vGPCR were further able to promote tumor formation by cells expressing KSHV latent genes, suggestive of a cooperative role among viral genes in the promotion of Kaposi's sarcomagenesis. PMID- 12559174 TI - The RAG-1/2 endonuclease causes genomic instability and controls CNS complications of lymphoblastic leukemia in p53/Prkdc-deficient mice. AB - Double-strand DNA breaks (DSB) induce chromosomal translocations and gene amplification in cell culture, but mechanisms by which DSB cause genomic instability in vivo are poorly understood. We show that RAG-1/2-induced DSB cause IgH/c-Myc translocations in leukemic pro-B cells from p53/Prkdc-deficient mice. Strikingly, these translocations were complex, clonally heterogeneous and amplified. We observed reiterated IgH/c-Myc fusions on dicentric chromosomes, suggesting that amplification occurred by repeated cycles of bridge, breakage and fusion. Leukemogenesis was not mitigated in RAG-2/p53/Prkdc-deficient mice, but leukemic pro-B cells lacked IgH/c-Myc translocations. Thus, global genomic instability conferred by p53/Prkdc disruption efficiently transforms pro-B cells lacking RAG-1/2-induced DSB. Unexpectedly, RAG-2/p53/Prkdc-deficient mice also developed leptomeningeal leukemia, providing a novel spontaneous model for this frequent complication of human lymphoblastic malignancies. PMID- 12559175 TI - AURORA-A amplification overrides the mitotic spindle assembly checkpoint, inducing resistance to Taxol. AB - The serine-threonine kinase gene AURORA-A is commonly amplified in epithelial malignancies. Here we show that elevated Aurora-A expression at levels that reflect cancer-associated gene amplification overrides the checkpoint mechanism that monitors mitotic spindle assembly, inducing resistance to the chemotherapeutic agent paclitaxel (Taxol). Cells overexpressing Aurora-A inappropriately enter anaphase despite defective spindle formation, and the persistence of Mad2 at the kinetochores, marking continued activation of the spindle assembly checkpoint. Mitosis is subsequently arrested by failure to complete cytokinesis, resulting in multinucleation. This abnormality is relieved by an inhibitory mutant of BUB1, linking the mitotic abnormalities provoked by Aurora-A overexpression to spindle checkpoint activity. Consistent with this conclusion, elevated Aurora-A expression causes resistance to apoptosis induced by Taxol in a human cancer cell line. PMID- 12559176 TI - Integrin-mediated targeting of drug delivery to irradiated tumor blood vessels. AB - The objective of this study was to target drug delivery to radiation-induced neoantigens, which include activated receptors within the tumor vasculature. These responses include posttranslational changes in pre-existing proteins, which can be discovered by phage-displayed peptide libraries administered to mice bearing irradiated tumors. Phage-displayed peptides recovered from irradiated tumors included the amino acid sequence RGDGSSV. This peptide binds to integrins within the tumor microvasculature. Immunohistochemical staining of irradiated tumors showed accumulation of fibrinogen receptor alpha(2b)beta(3) integrin. We studied tumor targeting efficiency of ligands to radiation-induced alpha(2b)beta(3). Radiopharmaceuticals were localized to irradiated tumors by use of alpha(2b)beta(3) ligands conjugated to nanoparticles and liposomes. Fibrinogen conjugated nanoparticles bind to the radiation-activated receptor, obliterate tumor blood flow, and significantly increase regression and growth delay in irradiated tumors. Radiation-guided drug delivery to tumor blood vessels is a novel paradigm for targeted drug delivery. PMID- 12559178 TI - Histone modifications and silencing prior to DNA methylation of a tumor suppressor gene. AB - We attempted to answer two central questions about epigenetic silencing of the tumor suppressor gene p16(INK4a) in this study: (1) whether the maintenance of associated histone modifications is dependent on DNA methylation and (2) whether such histone modifications can occur prior to DNA methylation. By coupling chromatin immunoprecipitation with gene targeting and the analysis of specific alleles, we found that elimination of DNA methylation from a p16(INK4a) allele resulted in profound changes in surrounding histones. After continued passage of such cells, methylation of histone H3 lysine-9 occurred in conjunction with re silencing in the absence of DNA methylation. These results have important implications for understanding the biochemical events underlying the silencing of tumor suppressor genes and the resultant growth suppression. PMID- 12559177 TI - Loss of pVHL is sufficient to cause HIF dysregulation in primary cells but does not promote tumor growth. AB - Inactivation of the von Hippel-Lindau (VHL) gene is associated with the development of highly vascularized tumors. pVHL targets the alpha subunits of hypoxia inducible factor (HIF) for ubiquitin-mediated degradation in an oxygen dependent manner. Although pVHL-deficient tumor cell lines demonstrate constitutive stabilization and activation of HIF, it has yet to be shown that loss of murine Vhl alone is sufficient to dysregulate HIF. We utilized a genetic approach to demonstrate that loss of Vhl is sufficient not only to stabilize HIF alpha subunits under normoxia, but also fully activate HIF-mediated responses. These studies have implications for the hierarchy of signaling events leading to HIF stabilization, nuclear translocation, and target gene expression. We further demonstrate that loss of murine Vhl does not promote teratocarcinoma growth, indicating that other genetic changes must occur to facilitate Vhl-mediated tumorigenesis. PMID- 12559179 TI - Standard vs. radical pancreaticoduodenectomy for periampullary adenocarcinoma: a prospective, randomized trial evaluating quality of life in pancreaticoduodenectomy survivors. AB - This study was designed to assess the health-related quality of life (QOL) of patients who had been randomly assigned to either standard or radical pancreaticoduodenectomy for periampullary adenocarcinoma. Pancreaticoduodenectomy has been performed in increasing numbers for periampullary adenocarcinoma. The appropriate extent of resection (standard vs. radical [extended]) remains controversial, particularly as concerns survival benefit. Past reports comparing standard vs. radical resection have suggested that the more extensive resection is attended by negative functional outcomes (diarrhea and weight loss) and poorer QOL, diminishing the impact of any possible survival advantage of the radical resection. A prospective, randomized single-institution trial comparing standard pancreaticoduodenectomy (pylorus preservation preferred) to radical pancreaticoduodenectomy (including distal gastrectomy and retroperitoneal lymphadenectomy) evaluated 299 patients with periampullary adenocarcinoma between April 1996 and June 2001. A standard Functional Assessment of Cancer Therapy Hepatobiliary (FACT-Hep) QOL survey designed for hepatobiliary cancer was sent to 150 of these patients surviving pancreaticoduodenectomy. QOL and functional status were assessed via a series of subscale scores for physical, social, emotional, and functional well-being. A total of 105 QOL surveys (70%) were returned and analyzed, with 55 of the patients having been randomized to the standard group and 50 to the radical group. The patients were evaluated at a mean of 2.2 years after pancreaticoduodenectomy. The two groups were statistically similar with regard to multiple parameters including age at operation (64.6 years), race, intraoperative blood transfusions, pathologic diagnosis and staging, and perioperative complications. The radical group had a significantly higher percentage of men (66% vs. 44%; P = 0.02), a longer operative time (369 minutes vs. 327 minutes; P < 0.001), and a longer postoperative length of hospital stay (13.6 days vs. 10.1 days; P < 0.01). The FACT-Hep total QOL scores were similar between the standard and radical groups: 143.5 vs. 147.3, respectively. Additionally, the individual FACT-G subscale scores evaluating physical (22.1 vs. 23.3), social (24.5 vs. 24.4), emotional (19.2 vs. 19.6), and functional well-being (20.6 vs. 22.4) were comparable between the standard and radical groups. Subgroup analyses based on pathologic diagnosis (pancreatic, ampullary, distal bile duct, etc.) failed to reveal any differences in QOL assessment between the standard and radical pancreaticoduodenectomy groups. Finally, QOL measures were similar when comparing time since operation (<2 years' follow-up vs. >2 years' follow-up) and age (< or =65 years vs. >65 years). This is the largest report comparing QOL assessment in survivors of pancreaticoduodenectomy randomized between standard and radical resection. These data demonstrate no differences in long-term QOL between standard and radical resection. These results imply that no negative long-term QOL measures are associated with radical pancreaticoduodenectomy (as performed in this study) for periampullary adenocarcinoma. PMID- 12559180 TI - Mucus is a predictor of better prognosis and survival in patients with intraductal papillary mucinous tumor of the pancreas. AB - The aim of our study was to examine the case histories of patients with intraductal papillary mucinous tumor (IPMT) treated with resection to determine predictors of prognosis. Between 1989 and 2000, all patients treated with pancreatic resection for IPMT (n = 63) were analyzed. The diagnosis of IPMT was made using the surgical specimen and the World Health Organization definition. Predictors were determined using univariate and multivariate analysis. The pathologic findings were benign (n = 30), carcinoma in situ (CIS; n = 5), and invasive carcinoma (n = 28). After univariate analysis, predictors of malignancy (invasive plus CIS) were jaundice (odds ratio = 10.32), elevated serum CA19-9 (odds ratio = 15.0), any abnormal liver function test (odds ratio = 7.69), and p53 overexpression. The only predictor of benign disease was gross mucus observed during endoscopy (odds ratio = 4.35). After multivariate analysis, predictors of malignancy were any abnormal liver function test (odds ratio = 5.09) and p53 overexpression, whereas the only predictor of benign disease was still gross mucus (odds ratio = 5.88). Actuarial 3- and 5-year survival for benign disease was 95% and 83% and for malignant disease 52% and 44%, respectively (P = 0.0048). Survival curves also favored p53-negative tumors vs. p53-positive tumors (P = 0.0055). In the 33 patients with malignant disease (mean follow-up time = 35 months), the presence of gross mucus was a predictor of prolonged survival after univariate and multivariate analysis (odds ratio = 4.34 and 4.55, respectively), whereas alcohol abuse was a predictor of poor survival (odds ratio = 3.41 and 3.60, respectively). Gross mucus observed during endoscopy is a predictor of benign IPMT and, within the group with malignant IPMT; the presence of gross mucus was associated with better survival. Survival was also strongly associated with either benign IPMT or negative staining for p53 overexpression. PMID- 12559181 TI - Regulation of Kupffer cell TNF gene expression during experimental acute pancreatitis: the role of p38-MAPK, ERK1/2, SAPK/JNK, and NF-kappaB. AB - We have demonstrated that Kupffer cell-derived tumor necrosis factor (TNF) mediates pancreatitis-associated liver injury. The aim of this study was to determine the role of p38 mitogen-activated protein kinase (MAPK), extracellular stress-related kinase 1/2 (ERK1/2), stress-activated protein kinase/c-Jun N terminal kinase (SAPK/JNK), and nuclear factor-kappaB (NF-kappaB) in TNF gene expression within Kupffer cells. TNF and TNF-mRNA were measured in rat livers perfused with elastase. TNF, TNF-mRNA, NF-kappaB activation, and phosphorylated p38-MAPK, SAPK/JNK, and ERK1/2 were determined in Kupffer cells treated with elastase. Elastase increased TNF and upregulated TNF-mRNA in livers (P<0.03) and Kupffer cells (P<0.001). Phosphorylated p38-MAPK, SAPK/JNK, and ERK1/2 and activated NF-kappaB were detected in Kupffer cells at 7 minutes; at 60 minutes, TNF-mRNA peaked and NF-kappaB returned to baseline, whereas all three kinases remained activated. Gadolinium inhibited elastase-induced upregulation of TNF mRNA (P < 0.001), TNF production (P<0.001), and attenuated SAPK/JNK, as well as ERK1/2, but not p38-MAPK. Both UO126 and SB203580 significantly inhibited elastase-induced upregulation of TNF-mRNA and TNF production (P<0.001), but only UO126 inhibited activation of NF-kappaB. It was concluded that pretranscriptional regulation of TNF gene expression in Kupffer cells follows an orderly activation of p38-MAPK, ERK1/2, and SAPK/JNK that may not converge on NF-kappaB. The seemingly limited duration of NF-kappaB activation may be important in "switching off" the cytokine cascade during acute pancreatitis. PMID- 12559183 TI - Restoring apoptosis in pancreatic cancer cells by targeting the nuclear factor kappaB signaling pathway with the anti-epidermal growth factor antibody IMC-C225. AB - We have previously demonstrated that RelA is constitutively activated in the majority of human pancreatic cancers and plays an important role in tumorigenesis and metastasis. The antiapoptotic gene bcl-xl is a downstream target of RelA, and regulation of bcl-xl transcription is mediated directly by the nuclear factor kappaB (NF-kappaB) binding sites present in the upstream promoter element of the bcl-xl gene. In this study we investigated the effects of inhibition of epidermal growth factor receptor (EGFR) signaling pathway with the anti-EGFR monoclonal antibody IMC-C225 on constitutive NF-kappaB activation and regulation of apoptosis-related genes in human pancreatic cancer cells. We found that activation of EGFR can be blocked with the anti-EGFR antibody IMC-C225 in the human pancreatic cancer cell line MDA Panc-28, leading to a marked decrease in constitutive NF-kappaB DNA binding activity. Our data also suggest that downregulation of NF-kappaB DNA binding activity by IMC-C225 leads to a decrease in bcl-xl and bfl-1 expression. Therefore, targeting the NF-kappaB signaling pathway with an anti-EGFR antibody may be one strategy to restore apoptosis in human pancreatic cancer cells, thereby enhancing the effect of chemotherapy and radiation therapy. PMID- 12559184 TI - Clinical outcomes after laparoscopic antireflux surgery in patients with and without preoperative endoscopic esophagitis. AB - A wide spectrum of endoscopic findings exists in patients with gastroesophageal reflux disease (GERD). This study compared clinical outcomes after laparoscopic antireflux surgery (LARS) in patients who had GERD with and without preoperative endoscopic esophagitis. From 1992 to 2001, a total of 414 patients who underwent LARS with 6 months or more of follow-up were prospectively entered into a database. Among these patients, 84 (20%) had no endoscopic evidence of esophagitis on preoperative endoscopy (group 1), whereas 330 (80%) did have esophagitis (group 2). Perioperative outcomes, GERD symptom relief, and the use of acid-reducing medication were assessed. Preoperative DeMeester scores in groups 1 and 2 were 44 +/- 29 and 61 +/- 62 (P < 0.05) and mean esophageal peristaltic amplitude was 86 +/- 32 mm Hg vs. 60 +/- 42 mm Hg, respectively (P < 0.05). Although procedure time was significantly shorter in group 1, other perioperative outcomes were similar. At postoperative follow-up, the use of proton pump inhibitors was reduced in both groups (86% to < or =14%). With the exception of postoperative dysphagia, there was no difference in GERD symptom relief between groups 1 and 2. The presence or absence of preoperative esophagitis has minimal effect on favorable symptomatic outcomes after LARS. Thus LARS is an effective treatment option for patients, irrespective of endoscopic evidence of esophagitis, leading to excellent symptom relief and a marked reduction in the use of proton pump inhibitors. PMID- 12559182 TI - Intestinal hypoperfusion contributes to gut barrier failure in severe acute pancreatitis. AB - Intestinal barrier failure and subsequent bacterial translocation have been implicated in the development of organ dysfunction and septic complications associated with severe acute pancreatitis. Splanchnic hypoperfusion and ischemia/reperfusion injury have been postulated as a cause of increased intestinal permeability. The urinary concentration of intestinal fatty acid binding protein (IFABP) has been shown to be a sensitive marker of intestinal ischemia, with increased levels being associated with ischemia/reperfusion. The aim of the current study was to assess the relationship between excretion of IFABP in urine, gut mucosal barrier failure (intestinal hyperpermeability and systemic exposure to endotoxemia), and clinical severity. Patients with a clinical and biochemical diagnosis of acute pancreatitis were studied within 72 hours of onset of pain. Polyethylene glycol probes of 3350 kDa and 400 kDa were administered enterally, and the ratio of the percentage of retrieval of each probe after renal excretion was used as a measure of intestinal macromolecular permeability. Collected urine was also used to determine the IFABP concentration (IFABP-c) and total IFABP (IFABP-t) excreted over the 24-hour period, using an enzyme-linked immunosorbent assay technique. The systemic inflammatory response was estimated from peak 0 to 72-hour plasma C-reactive protein levels, and systemic exposure to endotoxins was measured using serum IgM endotoxin cytoplasmic antibody (EndoCAb) levels. The severity of the attack was assessed on the basis of the Atlanta criteria. Sixty-one patients with acute pancreatitis (severe in 19) and 12 healthy control subjects were studied. Compared to mild attacks, severe attacks were associated with significantly higher urinary IFABP-c (median 1092 pg/ml vs. 84 pg/ml; P < 0.001) and IFABP-t (median 1.14 microg vs. 0.21 microg; P = 0.003). Furthermore, the control group had significantly lower IFABP-c (median 37 pg/ml; P = 0.029) and IFABP-t (median 0.06 microg; P = 0.005) than patients with mild attacks. IFABP correlated positively with the polyethylene glycol 3350 percentage retrieval (r = 0.50; P < 0.001), CRP (r = 0.51; P < 0.001), and inversely with serum IgM EndoCAb levels (r = -0.32; P = 0.02). The results of this study support the hypothesis that splanchnic hypoperfusion contributes to the loss of intestinal mucosal integrity associated with a severe attack of pancreatitis. PMID- 12559185 TI - The effect of chronic pain syndromes and psychoemotional disorders on symptomatic and quality-of-life outcomes of antireflux surgery. AB - Psychoemotional disorders (PED) and chronic pain syndromes (CPS) are common problems. Many patients with these disorders also suffer from gastroesophageal reflux disease (GERD). It is unclear how PED/CPS affect outcomes of antireflux surgery; therefore, the purpose of this study was to determine if PED/CPS adversely affects the results of surgical therapy for GERD. All patients referred for surgical therapy for GERD completed both the GERD-HRQL symptom severity instrument and the SF-36 generic quality-of-life instrument prior to surgery. To be candidates for surgery, patients must have symptomatic GERD and objective evidence of pathologic reflux by upper endoscopy, esophageal manometry and 24 hour pH monitoring. Patients underwent either laparoscopic or open Nissen or Toupet fundoplication. Six to 24 months postoperatively, patients were evaluated for satisfaction and quality-of-life. Ninety-three percent of control patients compared to 25% of PED/CPS patients were satisfied with surgery (P < 0.001). Dissatisfaction in PED/CPS patients was generally due to persistent or new somatic complaints. Median total GERD-HRQL scores improved for both groups, although postoperative scores were worse in the PED/CPS group. PED/CPS patients had significantly worse SF-36 scores both preoperatively and postoperatively compared to control patients. SF-36 scores improved in four of eight domains in control patients and none in the PED/CPS patients. In conclusion, PED/CPS patients are generally dissatisfied with antireflux surgery. Although some patients do benefit from surgery, careful patient selection is required. PMID- 12559186 TI - Laparoscopic paraesophageal hernia repair, a challenging operation: medium-term outcome of 116 patients. AB - Laparoscopic paraesophageal hernia repairs performed in 116 patients between 1992 and 2001 were prospectively analyzed. Perioperative outcomes were assessed and follow-up was performed under protocol. There were 85 female and 31 male patients who had a mean (+/- SD) age of 65 +/- 13 years and an American Society of Anesthesiology score of 2.3 +/- 0.6. All but two patients underwent an antireflux procedure. Gastropexy was performed in 48 patients, an esophageal lengthening procedure in six patients, and prosthetic closure of the hiatus in six patients. Major complications occurred in five patients (4.3%) with two postoperative deaths (1.7%). Mean follow-up was 30 +/- 25 months; 96 patients (83%) have been followed for more than 6 months. Among these patients, 73 (76%) are asymptomatic, 11 (11%) have mild symptoms, and 12 (13%) take antacid medications. Protocol barium esophagograms were obtained in 69% of patients at 6 to 12 months' follow up. Recurrence of hiatal hernia was documented in 21 patients (22% overall and in 32% of those undergoing contrast studies). Reoperation has been performed in three patients (3%). When only the patients with recurrent hiatal hernias are considered, 13 (62%) are symptomatic but only six (28%) require medication for symptoms. Laparoscopic paraesophageal hernia repair is generally safe, even in this high-risk group. This study confirms a relatively high incidence of recurrent hiatal abnormalities after paraesophageal hernia repair; however, most recurrent hiatal hernias are small and only 3% have required reoperation. Protocol esophagograms detect recurrences that are minimally symptomatic. Improved techniques must be devised to improve the long-term outcomes of laparoscopic paraesophageal hernia repair. PMID- 12559187 TI - Helicobacter pylori induces apoptosis in Barrett's-derived esophageal adenocarcinoma cells. AB - Helicobacter pylori may protect against the development of dysplasia in Barrett's epithelium of patients with gastroesophageal reflux disease. The aim of this study was to determine whether H. pylori preferentially induces apoptosis in Barrett's-derived cancer cells compared to normal cells. A Barrett's-derived adenocarcinoma cell line (OE33) was grown. H. pylori wild-type, isogenic vacA-, cagA(-), and picB-/cagE- mutant strains were grown on agar plates. Intact or sonicated bacteria were used to treat normal and OE33 cells for 24 hours, and Hoechst dye binding was performed to measure apoptosis. FAS protein expression was determined by Western immunoblotting. OE33 cells treated with intact H. pylori wild-type strains produced significant (P < 0.05) dose-dependent increases in apoptosis compared to normal esophageal cells. H. pylori wild-type and vacA- isogenic strains were more effective than cagA- and picB-/cage- isogenic strains in inducing apoptosis in OE33 cells. In OE33 cells, H. pylori sonicates produced lower levels of apoptosis than intact bacteria. Wild-type H. pylori strains increased Fas protein expression in OE33 cells at 18 hours. H. pylori induced apoptosis at a higher rate in the Barrett's-derived human esophageal adenocarcinoma cells than in normal esophageal cells. The H. pylori-induced apoptosis was primarily dependent on intact bacteria and the presence of the cagA and picB/cagE gene products. H. pylori-induced apoptosis may involve the Fas caspase cascade. PMID- 12559189 TI - Effect of aging on the adaptive and proliferative capacity of the small bowel. AB - Our society is aging at a rapid rate; the effects of aging on physiologic functions (e.g., small bowel adaptation) are poorly understood. The purpose of this study was to determine the ability of the aged small bowel mucosa to adapt after resection. Young (2-month-old) and aged (24-month-old) F344 rats underwent massive (70%) proximal small bowel resection (SBR) or sham operation; rats were killed at 9 or 16 days after surgery. The remnant small bowel and corresponding sham segments were harvested, weighed, and analyzed for DNA content and villus height. To determine whether the adaptive response after SBR could be enhanced, aged rats underwent SBR or sham operation and were treated with either neurotensin or saline solution (control). SBR resulted in adaptive hyperplasia in the remaining small bowel remnant in both young and aged rats at 9 and 16 days compared with sham animals. At 9 days, significant increases were noted in weight, villus height, and DNA content of the distal remnant in young and aged rats after SBR; the increases were similar in both young and aged rats. At 16 days, both young and aged rats displayed significant increases in remnant weight after SBR. Administration of neurotensin increased the weight of the remnant intestine in aged rats after SBR compared with saline treatment. Our findings demonstrate that aged small bowel mucosa exhibits a proliferative and adaptive capacity in response to SBR that was similar to that of the young animals. In addition, neurotensin administration enhanced the normal adaptive response of the small bowel in aged rats, providing further evidence that neurotensin may be therapeutically useful to augment mucosal regeneration in the early periods after massive SBR. PMID- 12559188 TI - Apoptotic and proliferative indexes in esophageal cancer: predictors of response to neoadjuvant therapy [corrected]. AB - Altered expression of the genes that control apoptosis and proliferation may influence the response of cancer cells to cytotoxic agents. The primary aim of this study was to determine the role of the novel antiapoptotic and cell cycle gene, survivin, in apoptotsis and proliferation in esophageal cancer and to evaluate whether the survivin, p53, and bcl-2 status were able to predict a patient's response to neoadjuvant therapy. A total of 104 patients with esophageal tumors were studied. Tumor tissue was immunostained for survivin, p53, and bcl-2 proteins. Proliferative and apoptotic activity was measured using ki-67 immunohistochemical analysis and the TUNEL method, respectively. Forty-eight patients whose pretreatment biopsies were analyzed received neoadjuvant chemoradiation therapy or chemotherapy followed by surgery. Outcome was graded as a complete response, a partial response, or no response according to the results of histologic examination and CT imaging. Expression of survivin was found to correlate significantly with the proliferative index but not the apoptotic index. Patients who received neoadjuvant treatment were more likely to achieve a complete response if their tumors had high proliferative activity, and p53 positive tumors were more likely to contain residual tumor after treatment. In conclusion, survivin expression appears to foster proliferative activity in esophageal cancer, and tumors with a high proliferative index or a functioning p53 gene are more responsive to neoadjuvant chemoradiation therapy. PMID- 12559190 TI - Evaluation of porcine-derived small intestine submucosa as a biodegradable graft for gastrointestinal healing. AB - High-risk anastomoses in the gut may benefit from the application of a synthetic reinforcement to prevent an enteric leak. Recently a porcine-derived small intestine submucosa (SIS) was tested as a bioscaffold in a number of organ systems. The aim of this study was to evaluate the effectiveness of SIS in stimulating healing in the stomach. Twelve rats underwent surgical removal of a full-thickness gastric defect (1 cm) and subsequent repair with a double-layer patch of porcine-derived SIS. The graft was secured with interrupted sutures placed within 1 mm of the edge of the graft. After 21 days, the animals were killed and their stomachs harvested for histologic examination. Cross sections were processed for paraffin embedding and 4-micron sections were stained with hematoxylin and eosin. All animals survived, gained weight, and demonstrated no signs of peritonitis over the 3-week postoperative period. On postmortem examination, the defect was completely closed in all animals by granulation tissue and early fibrosis. Although most of the luminal surface of the grafted areas remained ulcerated, early regeneration of normal gastric mucosa was seen at the periphery of the defect. SIS may act as an effective scaffolding agent for intestinal mucosa and may offer protection in high-risk anastomoses. PMID- 12559191 TI - Bariatric surgery for severely obese adolescents. AB - A 1991 National Institutes of Health Consensus Conference concluded that severely obese adults could be eligible for bariatric surgery if they had a body mass index (BMI) > or =35 kg/m(2) with or > or =40 kg/m(2) without obesity comorbidity. It was thought at that time that there were inadequate data to support bariatric surgery in severely obese adolescents. An estimated 25% of children in the United States are obese, a number that has doubled over a 30-year period. Very little information has been published on the subject of obesity surgery in adolescents. Therefore we reviewed our 20-year database on bariatric surgery in adolescents. Severely obese adolescents, ranging from 12 to less than 18 years of age, were considered eligible for bariatric surgery according to the National Institutes of Health adult criteria. Gastroplasty was the procedure of choice in the initial 3 years of the study followed by gastric bypass, which was found to be significantly more effective for weight loss in adults. Distal gastric bypass (D-GBP) was used in extremely obese patients (BMI > or =60 kg/m(2)) before 1992 and long-limb gastric bypass (LL-GBP) was used for superobese patients (BMI > or =50 kg/m(2)) after 1992. Laparoscopic gastric bypass was used after 2000. Thirty-three adolescents (27 white, 6 black; 19 females, 14 males) underwent the following bariatric operations between 1981 and June 2001: horizontal gastroplasty in one, vertical banded gastroplasty in two, standard gastric bypass in 17 (2 laparoscopic), LL-GBP in 10, and D-GBP in three. Mean BMI was 52 +/- 11 kg/m(2) (range 38 to 91 kg/m(2)), and mean age was 16 +/- 1 years (range 12.4 to 17.9 years). Preoperative comorbid conditions included the following: type II diabetes mellitus in two patients, hypertension in 11, pseudotumor cerebri in three, gastroesophageal reflux in five, sleep apnea in six, urinary incontinence in two, polycystic ovary syndrome in one, asthma in one, and degenerative joint disease in 11. There were no operative deaths or anastomotic leaks. Early complications included pulmonary embolism in one patient, major wound infection in one, minor wound infections in four, stomal stenoses (endoscopically dilated) in three, and marginal ulcers (medically treated) in four. Late complications included small bowel obstruction in one and incisional hernias in six patients. There were two late sudden deaths (2 years and 6 years postoperatively), but these were unlikely to have been caused by the bariatric surgical procedure. Revision procedures included one D-GBP to gastric bypass for malnutrition and one gastric bypass to LL-GBP for inadequate weight loss. Regain of most or all of the lost weight was seen in five patients at 5 to 10 years after surgery; however, significant weight loss was maintained in the remaining patients for up to 14 years after surgery. Comorbid conditions resolved at 1 year with the exception of hypertension in two patients, gastroesophageal reflux in two, and degenerative joint disease in seven. Self-image was greatly enhanced; eight patients have married and have children, five patients have completed college, and one patient is currently in college. Severe obesity is increasing rapidly in adolescents and is associated with significant comorbidity and social stigmatization. Bariatric surgery in adolescents is safe and is associated with significant weight loss, correction of obesity comorbidity, and improved self-image and socialization. These data strongly support obesity surgery for those unfortunate individuals who may have difficulty obtaining insurance coverage based on the 1991 National Institutes of Health Consensus Conference statement. PMID- 12559192 TI - Importance of response to neoadjuvant chemotherapy in patients undergoing resection of synchronous colorectal liver metastases. AB - The purpose of this study was to compare the treatment and outcome in patients referred for staged resection of synchronous colorectal liver metastases. The records of patients who had undergone colon or rectal resection and were then referred for evaluation of clinically resectable synchronous liver metastases between January 1995 and January 2000 were reviewed. Comparisons were made between patients who did not receive neoadjuvant chemotherapy and had exploratory operations after recovery from colon resection and patients who did receive chemotherapy before liver resection. A total of 106 patients were treated during the 5-year period. Neoadjuvant chemotherapy was given to 52 of the patients; in 29 of them the disease did not progress, but in 17 patients the disease progressed while they were receiving treatment. Median follow-up was 30 months. Patient- and tumor-related variables were similar between groups. Five-year survival was statistically similar between patients who did and those who did not receive neoadjuvant chemotherapy (43% vs. 35%, P = 0.49). Patients within the neoadjuvant group whose disease did not progress while they were receiving chemotherapy experienced significantly improved survival as compared to patients who did not receive chemotherapy (85% vs. 35%, P = 0.03). In the setting of synchronous colorectal liver metastases, the response to neoadjuvant chemotherapy may be a prognostic indicator of survival and may assist in the selection of patients for conventional or experimental adjuvant therapies. PMID- 12559193 TI - Diagnostic accuracy of endoscopic ultrasound-guided fine-needle aspiration in patients with presumed pancreatic cancer. AB - Endoscopic ultrasound (EUS)-guided fine-needle aspiration (FNA) of the pancreas allows the diagnosis of pancreatic cancer to be established without exploratory surgery. We reviewed our recent experience with EUS-FNA in patients with presumed pancreatic cancer and report the diagnostic accuracy and complications of this procedure. Data were reviewed from all patients who presented with CT evidence of a pancreatic mass or a malignant biliary stricture and underwent EUS-FNA at our institution between November 1, 1999, and October 1, 2001. Based on the findings of contrast-enhanced, multislice CT scanning, patients were categorized as having resectable, locally advanced, or metastatic disease. EUS-FNA was performed in 233 patients. A final diagnosis of cancer was established in 216 patients (93%), 15 patients (6%) were found to have benign disease, and the final diagnosis remains unknown in two patients (1%). The sensitivity, specificity, and accuracy of EUS FNA for diagnosis of a pancreatic malignancy were 91%, 100%, and 92%, respectively. For the 216 patients subsequently proven to have cancer, the results of EUS-FNA were diagnostic in 197 (91%); 96 (90%) of 107 patients with resectable disease, 62 (97%) of 64 with locally advanced disease, and 39 (87%) of 45 with metastatic disease. Four patients (2%) developed a clinically apparent complication that required hospital admission, including two patients who required surgery for duodenal perforation. There were no EUS-related deaths. We conclude that EUS-FNA can safely and accurately establish a cytologic diagnosis in patients with both early-stage and advanced pancreatic cancer. This enables consideration of all treatment options including protocol-based therapy. PMID- 12559194 TI - Lymphoplasmacytic sclerosing pancreatitis: inflammatory mimic of pancreatic carcinoma. AB - Lymphoplasmacytic sclerosing pancreatitis (LP) is a rare cause of benign mass lesions of the pancreas that can resemble adenocarcinoma. This study evaluates and classifies a series of patients with LP. Patients with benign pancreatic disease were identified from a prospective pancreatic database, and these cases were reviewed to identify patients with LP. Patients were subdivided into two groups: (1) classic LP, which included those patients who had all four of the characteristic histologic features of LP, including lymphoplasmacytic infiltration of the pancreas, interstitial fibrosis, periductal inflammation, and periphlebitis; and (2) intermediate LP, which included patients with at least two of these histologic findings. Patient demographics, pathologic and clinical features, and outcome were analyzed. From 1985 to 2001, a total of 1287 pancreatic resections were performed at our institution, of which 159 were for benign disease. Of these, 31 had pathologic features consistent with LP, and all of these patients had a presumed preoperative diagnosis of pancreatic carcinoma. Most of these patients presented with jaundice (n = 21) or abdominal pain (n = 7). In 29 of 31 patients, curative resection was possible. Of these, 28% (8/29) developed recurrence after resection: seven with jaundice and one with recurrent pancreatitis (median time to recurrence, 11 months; median follow up, 38 months). All patients with recurrent jaundice appeared to have biliary strictures at the time of direct cholangiography and no patient had malignancy. A review of the pathology reports identified 19 patients with classic LP and 12 patients with intermediate LP, and there was no difference between these two groups. LP is a rare cause of pancreatitis that is difficult to differentiate from carcinoma preoperatively. Patients with classic and intermediate LP appear to demonstrate a similar clinical behavior. Nearly one third of patients have a progressive course after resection, with 25% developing recurrent jaundice; thus close follow-up is mandatory for all patients. PMID- 12559195 TI - Modulators of ceramide metabolism sensitize colorectal cancer cells to chemotherapy: a novel treatment strategy. AB - Irinotecan is a first-line chemotherapeutic agent for patients with metastatic colorectal cancer (CRC). Response rates of less than 40% underscore the problem of treating CRC with irinotecan. Our studies have shown that chemosensitization correlates with high levels of ceramide, whereas resistance correlates with high levels of glucosylceramide (GlcCer). The purpose of this study was to characterize the role of ceramide in irinotecan-mediated CRC cell death. We used four human CRC cell lines to assess ceramide metabolism, cell viability, and apoptosis after treatment with irinotecan. Fumonisin B(1) (FB(1)) and 1-phenyl-2 palmitoylamino-3-morpholino-1-propanol (PPMP) were used to inhibit de novo ceramide synthesis and GlcCer production, respectively. L-threo dihydrosphingosine (safingol) was used to inhibit secondary proliferative pathways mediated by an atypical protein kinase C that is activated by ceramide. Irinotecan elicited dose- and time-dependent increases in ceramide, which preceded apoptosis. When FB(1) was added to irinotecan, CRC cell death was significantly decreased. A significant increase in intracellular levels of GlcCer also was noted after treatment with irinotecan. When GlcCer production was blocked by treating cells with PPMP in addition to irinotecan, ceramide levels increased to 228% of control values and cell death increased by 88%, compared to irinotecan alone. When irinotecan was combined with both PPMP and safingol, cell death was increased by 225% to 325%, compared to irinotecan lone. CRC cell death due to irinotecan is mediated, at least in part, by the de novo synthesis of ceramide. Blocking further metabolism of ceramide can enhance this cytotoxicity. Targeting ceramide pathways is a novel strategy for the treatment of patients with CRC. PMID- 12559196 TI - Epidermal growth factor activation of intestinal glutamine transport is mediated by mitogen-activated protein kinases. AB - Glutamine is an essential nutrient for gut functions, but the regulation of its uptake by intestinal mucosal cells is poorly understood. Given the pivotal role of epidermal growth factor (EGF) in regulating gut metabolism, growth, and differentiation, this in vitro study was designed to investigate the intracellular signaling pathways involved in the regulation of EGF-mediated intestinal glutamine transport in intestinal epithelia. Continuous incubation with EGF (>30 hours, 100 ng/ml) stimulated glutamine transport activity across intestinal epithelial Caco-2 cell apical membrane. Exposure to EGF for 48 hours resulted in an increase in transport activity (50%) and glutamine transport system B gene ATB(0) mRNA levels (ninefold). EGF stimulated glutamine transport activity by increasing the glutamine transporter maximal velocity (V(max)) without altering the transporter apparent affinity (K(m)). Furthermore, EGF stimulated both intracellular protein kinase C and mitogen-activated protein kinase MEK1/2 activities. The EGF-stimulated glutamine transport activity was attenuated individually by the specific protein kinase C inhibitor chelerythrine chloride and the mitogen-activated protein kinase MEK1 inhibitor PD 98059. These data suggest that EGF activates glutamine transport activity across intestinal epithelial membrane via a signaling mechanism that involves activation of protein kinase C and the mitogen-activated protein kinase MEK1/2 cascade. EGF activates glutamine transport via alterations in transporter mRNA levels and the number of functional copies of transporter units. PMID- 12559197 TI - Regional and muscle layer variations in cholinergic nerve control of the rat myometrium during the oestrous cycle. AB - To determine regional and muscle layer differences in the cholinergic nerve control of uterine activity, functional and immunohistochemical experiments were carried out on the cervix, and circular and longitudinal muscle from the caudal and rostral uterine horn in cyclic rats. During oestrus, in vitro electrical field stimulation evoked contractions in the cervix and myometrium of the caudal horn, predominantly in circular muscle layer. All evoked responses were tetrodotoxin-sensitive and completely abolished by atropine, thus were cholinergic nerve-mediated. In contrast, no electrical field stimulation-induced contraction occurred in either the circular or longitudinal muscle from the rostral uterus. Concentration-response curves for carbachol showed that muscarinic receptor-mediated contractions occurred in all uterine regions and muscle layers during oestrus. Immunohistochemistry for the cholinergic nerve marker, vesicular acetylcholine transporter showed that the predominance of the acetylcholine-dependent contractions in circular muscle preparations were related to a layer-specific distribution of cholinergic nerve fibres, abundant in the circular muscle but scarce in the longitudinal muscle layer. In addition, the absence of electrical field stimulation-evoked acetylcholine-dependent contractions in the rostral uterus was correlated to a marked decrease in the density of cholinergic fibres along the caudo-rostral axis of the organ. In the uterus from diestrus rats, contractions were not elicited in response to electrical field stimulation in the cervix and circular or longitudinal muscle from the caudal as well as rostral uterine horn. Addition of cumulative doses of carbachol failed to increase in a concentration-dependent manner the frequency and amplitude of contractions in the cervix and myometrial layers from either the caudal and rostral uterine horn. The distribution and density of cholinergic nerve fibres along the uterus and between the muscle layers did not differ from the oestrus stage. We conclude that the cholinergic nerve control of uterine activity is layer-specific and predominant in the caudal uterine horn and the cervix. Impairment of this nerve control from oestrus to diestrus stages occurred in relation to a decrease in the myometrial sensitivity to muscarinic stimulation, not to a decrease in the density of cholinergic innervation. PMID- 12559198 TI - Involvement of a purinergic pathway in the sympathetic regulation of motility in rat ileum. AB - We investigated extrinsic neuronal regulation of intestinal motility. The mesenteric nerve stimulation (MNS; duration 0.5 ms, 10 Hz for 30 s) evoked relaxation in the longitudinal muscle direction of the isolated rat ileum. The MNS-induced relaxation was abolished by guanethidine (2 microM) or propranolol (10 microM), but was not affected by prazosin (10 microM), rauwolscine (10 microM), hexamethonium (100 microM) or capsaicin (1 microM). Exposure to a high concentration (100 microM) of ATP (ATP-desensitization) or ADP (ADP desensitization) reduced the MNS-induced relaxation to 44.7% or 32.5% of the control (P<0.01), respectively. P2 purinoceptor antagonists [suramin (100 microM) and reactive blue-2 (RB-2, 50 microM)] or small conductance Ca(2+)-activated K(+) channel blocker, apamin (0.5 microM), significantly decreased the relaxation to 54.4% and 25.6% or 19.4% of the control (P<0.01), respectively, whereas selective P2Y(1) purinoceptor antagonist MRS2179 (10 microM) failed to affect the relaxation. Furthermore, exogenous ATP (1 microM) or ADP (1 microM) elicited relaxation in the rat ileum, which was almost abolished by reactive blue-2 (50 microM, 9.1% of control remained, P<0.05). In contrast, relaxation induced by noradrenalin (10 microM) was not antagonized by ATP-desensitization, apamin (0.5 microM) or reactive blue-2 (50 microM). From the present results, we conclude that noradrenergic sympathetic nerves might regulate intestinal motility mediated through a purinergic inhibitory neuronal pathway in the rat small intestine. PMID- 12559199 TI - Attenuation by spinal cord stimulation of a nociceptive reflex generated by colorectal distention in a rat model. AB - The mechanisms underlying the cause and treatment of visceral pain of gastrointestinal origin are poorly understood. Previous clinical studies have shown that spinal cord stimulation (SCS) attenuates neuropathic and ischemic pain, and animal experiments have provided knowledge about probable physiological mechanisms. The goal of the present study was to investigate whether SCS influences colonic sensitivity in a conscious rat. A visceromotor behavioral response (VMR), induced by colorectal distention, was used to quantify the level of colonic sensitivity. Under anesthesia, an electrode (cathode) was placed on the dorsal surface of the spinal cord at L1. One week after implantation of the SCS electrode, the effects of stimulation delivered with different intensities (50 Hz, 0.2 ms for 30 min) on colonic sensitivity were determined. Nociceptive levels of colorectal distention (60 mm Hg for 10 min) induced an enhanced VMR quantified as an increased number of abdominal muscle contractions compared to controls in which the balloon catheter was inserted into the colorectal region but not distended. Colonic sensitization with acetic acid increased the VMR to innocuous levels of colorectal distention (30 mm Hg for 10 min). We found that SCS induced a significant depression of the VMR produced by colorectal distention in both normal rats and those with sensitized colons. The suppressive effect of SCS on colonic sensitivity suggests that SCS may have therapeutic potential for the treatment of visceral pain of gastrointestinal origin associated with abdominal cramping and painful abdominal spasms. PMID- 12559200 TI - Peripheral hyperosmolality reduces cardiac baroreflex sensitivity. AB - The effects of acute increases in peripheral osmolality on cardiac baroreflex sensitivity were tested in conscious, unrestrained rats by fitting values for arterial blood pressure and heart rate during acute pressor and depressor responses to a four-parameter, sigmoid logistic function curve. Cardiac baroreceptor reflex function curves were produced before and following 30 min intravenous (i.v.) infusion of 2.5 M NaCl, an equipressor concentration of phenylephrine, isotonic saline, or 2.5 M NaCl following central administration of an angiotensin-converting enzyme (ACE) inhibitor. Hypertonic saline infusion increased blood pressure, reduced heart rate and baroreflex sensitivity (gain and range), and shifted the baroreflex function curve to a higher operating blood pressure. These effects were prevented or attenuated by prior central administration of an ACE inhibitor. Isotonic volume expansion did not alter baroreflex responses. Finally, phenylephrine produced similar changes in blood pressure but did not alter cardiac baroreflex responses. These data demonstrate that acute increases in peripheral osmolality and/or sodium decrease cardiac baroreflex sensitivity through generation of central angiotensin II, independent of changes in extracellular fluid volume and blood pressure. PMID- 12559201 TI - Evaluation of the population of NADPH-diaphorase-stained and myosin-V myenteric neurons in the ileum of chronically streptozotocin-diabetic rats treated with ascorbic acid. AB - We investigated the effect of the ascorbic acid on the nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d)-stained and myosin-V myenteric neurons in the ileum of chronically diabetic rats. The study was performed 4 months after inducing experimental diabetes with streptozotocin. Diabetic rats showed increased (p<0.05) glycaemia and glycated haemoglobin. Three groups were compared, i.e., nondiabetic rats, diabetic rats and diabetic rats treated with ascorbic acid. Myosin-V immunohistochemistry and NADPH-d histochemistry were employed. We investigated the areas of 500 cell bodies of myosin-V neurons and of 500 NADPH-d-stained neurons from all groups. The quantitative analysis was performed by using an area of 8.96 mm(2) from each ileum. The two groups of diabetic rats and diabetic rats treated with ascorbic acid showed reduction in the number and an increased area of the myosin-V-immunostained myenteric neurons. In addition, we observed increased relative proportion of NADPH-d-stained neurons in diabetic rats and diabetic rats treated with ascorbic acid. However, the area of these neurons in the diabetic rats group was larger than those evidenced in the nondiabetic rats and diabetic rats treated with ascorbic acid. PMID- 12559202 TI - Effect of water immersion on renal sympathetic nerve activity and arterial baroreflex in conscious rabbits with heart failure. AB - Several studies have indicated an interaction between cardiopulmonary mechanoreflex and arterial baroreflex. However, the contribution of cardiopulmonary mechanoreflex to an abnormal arterial baroreflex in chronic heart failure (CHF) has not been fully investigated. We examined the effect of the activation of cardiopulmonary mechanoreceptors induced by head-out water immersion (WI) on the arterial baroreflex control of heart rate (HR) and renal sympathetic nerve activity (RSNA) in conscious rabbits with CHF induced by myocardial infarction. The arterial baroreflex sensitivity (BRS) of both HR and RSNA were decreased in CHF. WI induced a small decrease in RSNA in CHF compared to a sham-operated group (Sham), despite a similar increase in central venous pressure. WI did not affect BRS of HR or RSNA in either Sham or CHF. By averaging rectified RSNA recordings, we found that miniature RSNA in the control in CHF was higher than that in Sham. WI decreased the synchronized RSNA without changing miniature RSNA in either group. These results suggest that cardiopulmonary mechanoreflex control of RSNA is reduced in CHF, and that cardiopulmonary mechanoreflex has little effect on arterial baroreflex. An increase in miniature RSNA may reflect sympatho-excitation in CHF. PMID- 12559203 TI - Differential alterations in sympathetic neurotransmission in mesenteric arteries and veins in DOCA-salt hypertensive rats. AB - Sympathetic control of arteries and veins may be altered in hypertension. To test this hypothesis, constrictions of mesenteric arteries and veins caused by nerve stimulation and by norepinephrine (NE) and ATP were studied in vitro in tissues from deoxycorticosterone acetate (DOCA)-salt hypertensive and sham normotensive rats. In DOCA-salt arteries, the maximum neurogenic response was greater than that in sham arteries. The P2 receptor antagonist, pyridoxal-phosphate-6 azophenyl-2',4'-disulfonic acid (PPADS, 10 microM), greatly reduced neurogenic responses in sham but not DOCA-salt arteries. The alpha1-adrenergic receptor antagonist, prazosin (0.1 microM), inhibited responses in DOCA-salt but not sham arteries. Concentration-response curves for norepinephrine and ATP were similar in sham and DOCA-salt arteries, indicating that reactivity to sympathetic vasoconstrictor transmitters was not changed in DOCA-salt arteries. Neurogenic constrictions in sham and DOCA-salt veins were similar in amplitude, and they were completely blocked by prazosin. However, concentration-response curves for norepinephrine in DOCA-salt veins were right-shifted compared to those in sham veins. Cocaine (10 microM) and corticosterone (10 microM) caused a leftward shift in norepinephrine concentration-response curves in DOCA-salt but not sham veins. Norepinephrine content was decreased in DOCA-salt arteries and veins, and there was an increased norepinephrine transporter (NET) level in DOCA-salt veins. These data indicate that, in DOCA-salt hypertension, there is an increased norepinephrine release from sympathetic nerves associated with mesenteric arteries and veins. In arteries, this results in an increase in the amplitude of neurogenic constrictions. In veins, increased norepinephrine release maintains neurogenic constrictions in the presence of increased NET levels. PMID- 12559204 TI - Arithmetic calculation, deep inspiration or handgrip exercise-mediated pre operational active palmar sweating responses in humans. AB - We examined the effects of repetitive mental stimulation such as arithmetic calculations with sequential subtraction or physical tasks such as handgrip exercise and deep breathing on active palmar sweating responses in humans. Thirty three healthy, male and female volunteer students (20.4+/-2.1 years) participated in the present study. The responses of active palmar sweating were evaluated by using the newly developed ratemeter. The galvanic skin response (GSR) was also recorded in 10 out of 33 subjects. The oral explanation of the stimulation or tasks caused a rapid and wavy active palmar sweating response. The pre operational responses of active palmar sweating to the stimulation or tasks were also observed by the GSR recording. The mental stimulation- and physical tasks mediated pre-operational responses were significantly reduced by the trials. The mental stimulation or physical tasks also caused a rapid and oscillatory response of active palmar sweating during operation of the stimulation or tasks. The operation-mediated responses to physical tasks were observed ipsilaterally and contralaterally. The physical task-mediated responses were also reproducible, resulting in no habituation. In contrast, the operation-mediated responses to mental stimulation were reduced significantly by the trials, resulting in a marked habituation. The findings suggest that the mental stimulation- and physical tasks-mediated pre-operational responses of active palmar sweating obtained by using the newly developed ratemeter will make useful tests for evaluating neuronal activity of limbic system including amygdala, sympathetic sudomotor activity in the palmar skin and functional properties of the palmar sweat glands. PMID- 12559205 TI - Assessment of autonomic function in high level athletes by pupillometry. AB - Spectral analysis of heart rate variability has become a noninvasive standard method for assessment of autonomic nervous system activity in athletes. The effect of exercise training on autonomic regulation of pupillary light reflex is not known. The purpose of this study was to evaluate pupil autonomic function in athletes. We studied 46 highly trained athletes practicing gymnastics, swimming, long-distance running, soccer, and 51 healthy control subjects, using a portable infrared pupillometry. Five left pupil light response curves were recorded for each subject; the 485 pupillogram records were processed by a computer system. The following pupillometric parameters calculated were significantly higher (P<0.05) in runners than in controls: reflex amplitude (2.1 mm; 95% CI, 1.9-2.3 vs. 1.8 mm; 95% CI, 1.7-1.9), mean percent reflex amplitude of initial diameter (34%; 95% CI, 32-37 vs. 30%, 95% CI, 28-31) and mean time at which pupil redilated 75% of reflex amplitude (2.15 s; 95% CI, 1.99-2.31 vs. 1.86 s; 95% CI, 1.78-1.93). Sex, age, height, weight, body mass index and years of sports practice had no significant influence in the evaluated parameters. The results were consistent with an increased parasympathetic activity and a reduced sympathetic activity of pupillary light reflex in endurance-trained runners, supporting the hypothesis of a generalized "dysautonomy" associated with this type of training. PMID- 12559207 TI - Anaemia in heart failure: its diagnosis and management. PMID- 12559208 TI - The pharmacological management of heart failure: too many treatments? PMID- 12559210 TI - Pharmacotherapeutic approaches for decompensated heart failure: a role for the calcium sensitiser, levosimendan? AB - Although no universal definition exists, decompensated heart failure may be regarded as either a worsening of chronic heart failure or new-onset heart failure precipitated by an acute incident. Haemodynamic management of patients hospitalised with decompensated heart failure may include the administration of diuretics, vasodilators and positive inotropic agents. Until recently, these latter agents constituted the only drug class to produce a direct increase in stroke volume via enhanced myocardial contractility. However, despite their short term benefits, the clinical utility of inotropic agents is compromised by their potentially deleterious effects on calcium handling and oxygen consumption, resulting in an increased risk of serious ventricular arrhythmias and death. In contrast, calcium sensitisers enhance cardiac performance without affecting calcium movement and, therefore, are potentially associated with a reduced risk of rhythmic disturbances. These agents constitute a heterogeneous group of compounds with different affinities for calcium sensitisation. Levosimendan is a potent calcium sensitiser with vasodilating properties that has been shown to provide symptomatic and haemodynamic improvement with no increase in oxygen consumption. Calcium sensitisation is therefore emerging as a promising treatment approach in this challenging therapeutic area. PMID- 12559209 TI - OPTIME in CHF trial: rethinking the use of inotropes in the management of worsening chronic heart failure resulting in hospitalization. PMID- 12559211 TI - Plasma concentrations of interleukin-2 soluble receptor in mild ischaemic left ventricular dysfunction. PMID- 12559212 TI - Quantification of proinflammatory cytokines in the urine of congestive heart failure patients. Its relationship with plasma levels. AB - AIMS: Proinflammatory cytokines are important mediators for the development of heart failure and increased plasma levels of these cytokines have been reported in patients with this condition. The purpose of the study was to investigate whether urine, a non-invasively obtained biological sample, was an appropriate medium in which to measure the concentration of tumor necrosis factor-alpha (TNF alpha) and interleukin-6 (IL-6) in patients in the advanced stages of the disease. METHODS AND RESULTS: Thirty consecutive patients who had severe congestive heart failure (NYHA classes III and IV) and 30 matched healthy control subjects were enrolled. Plasma and the first urine of the day were collected and TNF-alpha and IL-6 were quantitatively analyzed by enzyme-linked immunosorbent assays. For every subject there were no differences in the amount of cytokine determined in plasma and urine. Both urine and plasma levels of IL-6 and TNF alpha were greater in heart failure patients than in controls. CONCLUSION: Our results show that plasmatic and urinary levels of proinflammatory cytokines did not differ significantly. Thus, urine may be a good milieu in which to study these cytokines and may have diagnostic, prognostic and therapeutic implications. PMID- 12559213 TI - Elevated serum levels of leptin and soluble leptin receptor in patients with advanced chronic heart failure. AB - Patients with chronic heart failure (CHF) have metabolic abnormalities, leading to a catabolic syndrome, with progressive loss of skeletal muscle in advanced stages of the disease. Leptin, the product of an obesity gene, has been associated with energy expenditure and weight regulation. The aim of this study was to assess serum levels of leptin and its soluble receptor in relation to exercise intolerance and neurohumoral activation in patients with CHF. We investigated 53 patients with CHF left ventricular ejection fraction (LVEF) 25+/ 1%, age 56.6+/-1.3 years, Maximal oxygen uptake (VO(2) max) 16.3+/-0.6 ml/min.kg) sub-classified according to peak oxygen consumption of > or 14 ml/min.kg and controls). Elevated levels of leptin correlated with an increased serum concentration of TNFalpha (r=0.749, P<0.01) in this subgroup of patients with CHF. We conclude that patients with advanced CHF show elevated serum levels of leptin and its soluble receptor. This finding indicates that leptin may participate in the catabolic state leading to the development of cardiac cachexia in the course of CHF. PMID- 12559214 TI - The frequency of systolic versus diastolic heart failure in an Egyptian cohort. AB - BACKGROUND: All factors predisposing for congestive heart failure (CHF), such as coronary artery disease (CAD), hypertension and diabetes are increasing in prevalence in Egypt. Despite this, no data about CHF in our country are available. AIMS: To study the relative contribution of systolic vs. diastolic heart failure in Egyptians and the prevalence of risk factors in this population, as well as their prognosis. METHODS: This was a retrospective study of patients with a diagnosis of CHF over a 3.5-year period in a general cardiology clinic. Demographic, ECG and echocardiographic data for left ventricular systolic and diastolic function were collected. The differential effect of systolic versus diastolic CHF was analyzed regarding hospitalization and mortality. RESULTS: After exclusion of valvular diseases, we found 155 patients diagnosed with heart failure, 102 patients (66%) had systolic heart failure, and 53 (34%) had diastolic heart failure. Mean age was 60+/-10 and 63+/-11 years, respectively (P=0.13). Systolic CHF patients had significantly more CAD, while those with diastolic failure were mostly hypertensives (P<0.01) for both. There was no significant difference in the incidence of diabetes mellitus, cerebrovascular accidents or atrial fibrillation between the two groups. Patients with systolic failure required more hospitalization, P<0.05, and had a mortality rate of 17.6% vs. 11.3% for patients with diastolic heart failure (P=0.3). CONCLUSION: Diastolic heart failure is present in one-third of cases of CHF in Egyptians. Hypertension is very common in this group. These patients require less hospitalization but have a similar mortality rate. PMID- 12559215 TI - Neurohormonal effects of furosemide withdrawal in elderly heart failure patients with normal systolic function. AB - BACKGROUND: In heart failure patients, diuretics cause renin-angiotensin aldosterone system (RAS) activation, which may lead to increased morbidity and mortality despite short-term symptomatic improvement. AIM: To determine changes in RAS activation and clinical correlates following furosemide withdrawal in elderly heart failure patients without left ventricular systolic dysfunction. METHODS AND RESULTS: We performed clinical assessments and laboratory determinations of aldosterone, plasma renin activity (PRA), atrial natriuretic peptide (ANP), norepinephrine, and endothelin in 29 heart failure patients [aged 75.1+/-0.7 (mean+/-S.E.M.) years], before, 1 and 3 months after placebo controlled furosemide withdrawal. Recurrent congestion occurred in 2 of 19 patients withdrawn, and in 1 of 10 patients continuing on furosemide. Three months after withdrawal, PRA had decreased -1.61+/-0.71 nmol/l/h (P<0.05). Decreases in aldosterone levels did not reach significance (-0.17+/-0.38 nmol/l). The decreases in PRA after withdrawal correlated with decreases in systolic (r(s)=0.61, P=0.020) and diastolic blood pressure (r(s)=0.80, P=0.01). Successful withdrawal was associated with increases in norepinephrine (+0.58+/-0.22 nmol/l) and ANP (+3.5+/-1.3 pmol/l) (P<0.05) after 1 month, but these changes did not persist after 3 months. Endothelin levels did not change in both groups. CONCLUSION: Successful furosemide withdrawal in elderly heart failure patients causes persistent decreases in RAS activation. PMID- 12559216 TI - Prognostic value of serial cardiac assessment and familial screening in patients with dilated cardiomyopathy. AB - OBJECTIVES: This prospective study was performed to analyse whether routine clinical follow-up investigations at 12+/-6 months add to risk stratification and improve survival rates in patients with a first diagnosis of dilated cardiomyopathy (DCM). METHODS: Four hundred and eighty consecutive patients (mean age 53.4+/-12.3 years, 369 males, mean NYHA class 2.4+/-0.8) with invasively confirmed DCM were included and followed for 3.9+/-3.5 years. Patients were requested to adhere to a follow up investigation within 6-18 months either at the referring physicians or at our out patient department. Two hundred and eighty-one of the 480 patients presented for follow up which consisted of a detailed evaluation of symptoms, standardized physical examination, 12-lead electrocardiogram recording and echocardiography. Seventeen patients were lost for follow up, 182 did not seek specialized medical follow up. Patients outcome was assessed by structured telephone interviews. RESULTS: Independent predictors of death or transplantation at initial diagnosis were LV-ejection fraction <30% (P=0.0001, risk ratio 2.25), LV-end diastolic pressure >or=15 mmHg (P=0.002, risk ratio 2.0), age >or=54 years, (P=0.04, risk ratio 1.55), and presence of left bundle branch block (P=0.046, risk ratio 1.53). On follow up investigations only deterioration of clinical status by at least one NYHA-class (P=0.001, risk ratio 2.6) and new onset or worsening of mitral regurgitation (P=0.02, risk ratio 1.8), remained independent prognostic factors for cardiac death. Patients who presented for routine follow up revealed significant better 5-year survival rates (n=281, 70%) than those who did not (n=153, 55%, P=0.005). CONCLUSIONS: Routine clinical follow up investigations within 6-18 months after first diagnosis of DCM adds to risk stratification and improves survival rates. PMID- 12559217 TI - Transmitral pulsed-Doppler echocardiography is a more accurate technique compared with two-dimensional echocardiography using dobutamine, in patients with one vessel coronary artery disease. AB - To examine the effects of dobutamine on pulsed-Doppler left ventricular filling indices and its utility for evaluation of CAD we studied 14 patients with normal coronary arteries (Group 1) and 39 patients with significant CAD (>70% diameter stenosis). Patients with coronary artery disease (CAD) were divided into two groups: patients with one-vessel coronary disease (Group 2); and those with multivessel CAD (Group 3). After stopping cardioactive treatment, patients underwent incremental dobutamine stress (5, 10, 20, 30 and 40 microg/kg/min) during pulsed-Doppler interrogation of diastolic filling with simultaneous heart rate and blood pressure measurements. The following transmitral Doppler variables were measured at baseline and at peak-dose of dobutamine: peak early (E) and peak atrial (A) velocity; E/A ratio; acceleration time (AT) and deceleration time (DT) of E wave; isovolumic relaxation time (IVRT); and time-velocity integral (TVI). Two-dimensional echocardiography was performed to detect regional asinergy and analyzed using a 16 segment model. RESULTS: Normals and CAD patients showed comparable changes in heart rate and blood pressure (P=NS between groups). Intergroup analysis of the changes of transmitral flow showed the significant changes for these indices (P<0.001): E velocity (-2.78+/-10.04, 12.4+/-9.4 and 16.47+/-10.65 cm/s); AT of E wave (1.66+/-2.47, -5.2+/-1.38 and -4.66+/-2.39 m/s(2)); DT of E wave (-0.23+/-0.18, 0.2+/-0.2 and 0.2+/-0.28 m/s(2)); and TVI of transmitral flow (-1.26+/-0.7, 3.5+/-1.75 and 4.1+/-1.66 cm), respectively for Groups 1, 2 and 3. All other transmitral Doppler variables showed insignificant changes (P=NS) to dobutamine between groups. It is important that the significance of these changes were the same for patients with one-vessel and those with multivessel coronary disease. In conclusion, during dobutamine stress testing, patients with CAD, had an abnormal response of these transmitral Doppler indices: E wave; AT of E wave; DT of E wave; and the TVI of transmitral flow. The abnormal responses of these Doppler indices of left ventricular filling are more accurate markers of significant single vessel CAD than new wall motion abnormalities during conventional DSE. PMID- 12559219 TI - Angioplasty of residual stenosis after severe anteroseptal myocardial infarction: is it able to improve systolic function and to prevent cardiac failure? PMID- 12559218 TI - Effect of early revascularisation in cardiogenic shock complicating acute myocardial infarction. A single center experience. AB - BACKGROUND: Five to 10% of patients with acute myocardial infarction develop cardiogenic shock and the majority of these patients are expected to die within the first few weeks. In this study, we review our recent experience in the management of patients with cardiogenic shock complicating MI and examine the effect of early invasive revascularisation on mortality. METHODS: Thirty-six consecutive patients who developed cardiogenic shock less than 48 h after MI were retrospectively evaluated and divided into two treatment groups. One group received early invasive revascularisation (n=24) and the other group had no early invasive revascularisation, but received similar conventional intensive care medical treatment (n=12). RESULTS: Baseline characteristics and hemodynamic variables were similar in both groups. Apart from invasive revascularisation and the use of intra aortic balloon counterpulsation (IABP), treatment strategies did not differ between the two groups. Thirty-day mortality was 21% in the revascularised group of patients and 58% in the non-revascularised group (P<0.05). CONCLUSIONS: Our data support previous observations suggesting that an aggressive treatment strategy including early invasive revascularisation and IABP is associated with improved short and long-term survival in patients with cardiogenic shock. Since early revascularisation appears safe with a considerable treatment benefit, this approach must be considered in patients with short shock duration early after MI. PMID- 12559221 TI - Clinical trials update from the American Heart Association meeting: PROSPER, DIAL, home care monitoring trials, immune modulation therapy, COMPANION and anaemia in heart failure. AB - This article continues a series of reports on research developments of particular interest to those involved in the management of patients with heart failure. Summaries of the following trials, reported at the 75th Scientific Sessions of the American Heart Association held in Chicago, Illinois between 17th and 20th November 2002 are included: PROSPER; DIAL; home care monitoring trials; immune modulation therapy; COMPANION; and anaemia in heart failure. PMID- 12559220 TI - Prognosis of patients with chronic coronary artery disease and severe left ventricular dysfunction. The importance of myocardial viability. AB - BACKGROUND AND AIM: The choice of optimal treatment strategy in patients with coronary artery disease (CAD) and severe left ventricular (LV) dysfunction is often difficult. The aim of this study was to compare long-term results of patients with chronic CAD, severe heart failure and a defined scope of myocardial viability treated with coronary revascularization, heart transplantation, or kept on medical therapy. METHODS: From 1993 to 2000, viability evaluation using low dose dobutamine echocardiography was performed in 124 patients with CAD and LV ejection fraction 0.05 for each comparison). CONCLUSIONS: Biochemical markers of bone turnover are elevated in men receiving gonadotropin-releasing hormone agonist treatment but not in men receiving bicalutamide monotherapy. These observations suggest that bicalutamide monotherapy may maintain bone mineral density and prevent fractures. PMID- 12559283 TI - Influence of age, anthropometry, and hepatic and renal function on serum prostate specific antigen levels in healthy middle-age men. AB - OBJECTIVES: To test the hypothesis of a causal relationship between clinical parameters, including age, anthropometry, and hepatic or renal function tests and serum prostate-specific antigen (PSA) levels and to determine the predictors of high serum PSA concentrations in healthy middle-age men. METHODS: Between January 1999 and December 2000, 6005 healthy men 40 to 59 years old who visited our hospital for a routine health checkup were entered into the study. The association between the clinical parameters and a high serum PSA level (greater than 2.5, 3.0, 3.5, or 4.0 ng/mL) was studied in three groups: the 10% with low clinical parameters, the 10% with high clinical parameters, and the remainder as a reference group. RESULTS: The univariate logistic regression analysis indicated that high or low age, body weight, body mass index, creatinine, and creatinine clearance were significant factors in relation to serum PSA concentration compared with the reference group. In the multivariate model used, only older age was positively related to the serum PSA concentration. CONCLUSIONS: The results of anthropometry and hepatic and renal function tests do not influence the serum PSA level in this population. Our findings suggest that serum PSA may be a reliable marker in middle-age men without severe hepatic or renal disease. PMID- 12559284 TI - Mass screening for prostate cancer: a comparative study in Natori, Japan and Changchun, China. AB - OBJECTIVES: To study the natural background of prostate cancer in Japan and China, mass checks with prostate-specific antigen (PSA)-based screenings were performed using identical procedures in Natori, Japan and Changchun, China. METHODS: For 7 years (1995 to 2001) in Natori, Japan, and for 3 years (1998 to 2000) in Changchun, China, 2212 Japanese and 3566 Chinese men older than 55 years were mass checked by PSA-based screening (serum PSA cutoff value 4.1 ng/mL). RESULTS: The PSA-positive rates (PSA 4.1 ng/mL or greater) and cancer detection rates in the screened persons of Natori and Changchun were 8.5% and 5.2% (P <0.0005) and 2.1% and 0.8% (P <0.0001), respectively. When the number of cancer cases detected was adjusted to a 100% biopsy rate for men who were PSA positive in both cities, the cancer detection rate was estimated at 2.3% and 1.3% in Natori and Changchun, respectively. This difference was also significant (P <0.01). CONCLUSIONS: The results indicate that the percentage of PSA-positive men 55 years or older in Changchun was lower than that in Natori. The analysis of the results suggests that the prostate cancer incidence and prevalence in Changchun, China are lower than those in Natori, Japan. PMID- 12559285 TI - Estimating survival benefit in castrate metastatic prostate cancer: decision making in proceeding to a definitive phase III trial. AB - OBJECTIVES: In designing a Phase II trial, the acceptable clinical activity region for a new therapy is often developed using data from historically treated patients. This region incorrectly ignores the variability of this estimate, because the efficacy of the prior treatment lies somewhere around the estimate. The size of this interval is dependent on the sample size used. This report illustrates the use of a published method that accounts for this uncertainty and aids in the decision to proceed to a definitive trial. METHODS: A historical data set of low-risk patients with progressive castrate metastatic prostate cancer and a group of similar patients treated in a Phase II chemotherapy trial were used. The 1-year Kaplan-Meier estimate of survival was obtained for both. This approach uses the 75% upper confidence bound of the 1-year survival probability from the historical data set to define the lower limit of acceptable clinical activity. Use of this bound makes the approach more conservative, and hence the decision to proceed to a Phase III trial more difficult. RESULTS: In the low-risk historical patients, the 1-year Kaplan-Meier estimate of survival was 66.4% (75% upper confidence bound 71.0%). In the Phase II patients, the 1-year Kaplan-Meier estimate of survival was 89.5% (95% lower confidence bound 78.2%). CONCLUSIONS: A hypothesis test using the 75% upper confidence bound to define the lower limit of acceptable clinical activity demonstrates that the 1-year survival probability on Taxol/estramustine/carboplatin is greater than that of the historical population, and hence should be taken into a definitive trial. The design provides investigators increased confidence in making this decision. PMID- 12559286 TI - Does prophylactic breast irradiation prevent antiandrogen-induced gynecomastia? Evaluation of 253 patients in the randomized Scandinavian trial SPCG-7/SFUO-3. AB - OBJECTIVES: To examine the development of antiandrogen-induced gynecomastia and breast tenderness in the first 253 patients in a randomized Scandinavian trial (SPCG-7/SFUO-3) with a 12-month complete follow-up evaluation performed by both doctors and patients. METHODS: In this study, the treating doctor and patient decided whether prophylactic irradiation (RT) of the breast should be given to prevent antiandrogen-induced gynecomastia. At each visit, the doctor evaluated the occurrence of gynecomastia and breast tenderness. Questions about gynecomastia and breast tenderness were also included in the study quality-of life questionnaire (Prostate Cancer Symptom Scale). RESULTS: Mammary RT with mostly single fraction (12 to 15 Gy) electrons was given to 174 (69%) of the 253 evaluated patients. At the 1-year follow-up visit, the doctor evaluations indicated some form of gynecomastia in 71% and 28% (P <0.001) of the nonirradiated (no-RT) and irradiated (RT) patients, respectively. The patient evaluations at 1 year showed some form of breast enlargement in 78% and 44% (P <0.001) of the no-RT and RT patients, respectively. The doctors reported some form of breast tenderness at 1 year in 75% and 43% (P <0.001) of the no-RT and RT patients, respectively. The patient evaluations of breast tenderness show an expected significant increase in the RT arm at the 3-month follow-up, which was probably due to skin reactions. At 1 year, significantly more patients who marked "very much" on the Prostate Cancer Symptom Scale were seen in the no-RT group. A weak correlation between the doctors' and patients' detection of breast problems was observed. CONCLUSIONS: The results show that, with high significance, prophylactic RT of the breast decreases the risk of antiandrogen-induced gynecomastia and breast tenderness. PMID- 12559287 TI - Effect of anatomic, procedural, and dosimetric variables on urinary retention after permanent iodine-125 prostate brachytherapy. AB - OBJECTIVES: To correlate anatomic, procedural, and dosimetric parameters with the rate of intermittent self-catheterization (ISC). METHODS: The records of 402 patients with a median age of 69 years treated with 125I prostate seed implantation from 1996 to 2001 were reviewed for the use of ISC. The records were examined for the preimplant factors: prostate volume, use of androgen deprivation, and prostate length. The intraprocedural factor reviewed was the number of needles used. The following postimplant information was also collected: preimplant transrectal ultrasound-generated prostate volume/postimplant computed tomography-generated prostate volume ratio, V100, V150, V200, V300, V400, D90, and D100. Correlation was assessed using logistic regression analysis. RESULTS: Forty-four patients had to use ISC (10.9%). The mean and median duration of ISC was 11.9 and 6 weeks, respectively. From univariate analysis, prostate length and prostate volume were found to be statistically significant predictors of ISC after 125I prostate seed implantation with a P value of 0.0002 and 0.0042, respectively. With multivariate analysis, only prostate length was a statistically significant predictor of ISC use after 125I prostate seed implantation (P = 0.0095). CONCLUSIONS: Prostate length is an important predictor of ISC after 125I prostate seed implantation. PMID- 12559288 TI - Removal of urinary catheter on postoperative day 3 or 4 after radical retropubic prostatectomy. AB - OBJECTIVES: To determine the feasibility and safety of removing the urinary catheter on postoperative day (POD) 3 or 4 after radical retropubic prostatectomy (RRP). METHODS: Between January 2001 and August 2001, gravity cystography was performed on POD 3 or 4 after RRP by a single surgeon (H.L.) on 151 men. The urinary catheter was removed on POD 3 or 4 providing no extravasation was evident on cystography. Urinary catheters were replaced over a guidewire placed into the bladder under flexible cystoscopic guidance in cases of acute urinary retention (AUR). Continence was assessed at 3 months after RRP. RESULTS: Of the 151 cystograms performed on POD 3 or 4, 116 (76.8%) revealed no evidence of extravasation. The indwelling catheters were removed in 114 of these cases (98.3%). Twenty-two (19.3%) of the 114 men whose catheters were removed on POD 3 or 4 developed AUR within 48 hours of catheter removal, requiring catheter replacement. Two of these patients required repeated surgery secondary to complications related to AUR. At 3 months after RRP, 75% of the men whose catheters were removed on POD 3 or 4 required no pads or a single pad during a 24 hour interval, and 77.6% reported none or slight bother from incontinence. Of the 37 men whose catheters were removed on POD 7 or later, 1 patient (2.7%) developed AUR, and the catheter was replaced without complications. At 3 months after RRP, 65.7% of men whose catheters were removed on POD 7 or later required no pads or a single pad during a 24-hour interval, and 71.4% reported none or slight bother from incontinence. The incidence of anastomotic stricture in men whose catheters were removed on POD 3 or 4 and POD 7 or later was 12.1% and 22.6%, respectively. CONCLUSIONS: Most men will have a watertight anastomosis on POD 3 or 4 after RRP. Early catheter removal does not have a negative impact on continence or the rate of anastomotic strictures. Because of the high incidence of AUR, requiring replacement of the urinary catheter, and the potential for disruption of the anastomosis or bladder neck reconstruction, we currently recommend delaying catheter removal until POD 7 or later. PMID- 12559289 TI - Can total and transition zone volume of the prostate determine whether to perform a repeat biopsy? AB - OBJECTIVES: To compare the ability of total prostate (TP) and transition zone (TZ) volume to predict the outcome of a repeat prostate biopsy in patients with serum prostate-specific antigen (PSA) levels of 4 to 10 ng/mL. METHODS: A total of 1137 patients were included and underwent transrectal ultrasound-guided needle sextant and two transition zone biopsies of the prostate. All patients with a prior negative biopsy (benign prostatic tissue) underwent a repeat biopsy after 6 weeks. The TP and TZ volumes of the prostate were measured by transrectal ultrasonography. RESULTS: Of the 1137 patients, prostate cancer was diagnosed in 364 (32%), in 276 (24.2%) after the first biopsy and in 88 (7.7%) after the repeated biopsy. The TP and TZ volumes were larger in the patients with prostate cancer detected on the repeated biopsy (P <0.0001). Using a cutoff for TP volume of less than 20 cm3 and greater than 80 cm3 and for TZ volume of less than 9 cm3 and greater than 41 cm3 would have spared 7.1% and 10% of repeated biopsies, respectively. CONCLUSIONS: The probability for a positive repeat prostate biopsy increases in a logarithmic function for larger prostates, as well as for larger TP and, especially, for larger TZ volumes. The probability of finding prostate cancer on a repeat biopsy in prostates with small (less than 20 cm3) and large (greater than 79 cm3) TP, as well as in small (less than 9.3 cm3) and large (greater than 41 cm3) TZ volumes, was very low. Therefore, a repeat prostate biopsy within 6 weeks is unnecessary. These patients should be followed up by serial PSA determination. PMID- 12559290 TI - Influence of prostate volume in the detection of prostate cancer. AB - OBJECTIVES: To assess the influence of prostate volume on prostate cancer (CaP) detection in men who underwent repeated sextant transrectal ultrasound biopsy of the prostate. METHODS: Between September 1991 and September 2000, 4376 men underwent sextant transrectal ultrasound-guided biopsy of the prostate. Of the 4376 men, 556 underwent repeat biopsy because of persistent prostate-specific antigen elevation (greater than 4 ng/mL) and/or an abnormal digital rectal examination or suspicious pathologic findings. The percentage of CaP missed on the initial biopsy and detected on the repeat biopsy between arbitrary prostate volumes of less than 50 and 50 cm3 or greater and between less than 37.5 and 37.5 cm3 or greater, the median prostate volumes of men with CaP, were compared. Patient age, prostate-specific antigen level, digital rectal examination findings, and Gleason score in each volumetric cutoff group were also compared. RESULTS: CaP was detected in 22% of men who underwent a repeat biopsy. The percentage of CaP missed on the initial biopsy but subsequently detected on the repeat biopsy consistently increased as the volume increased. A statistically significant difference in the percentage of CaP not detected on the initial biopsy was found between prostate volumes of less than 50 and 50 cm3 or greater and between less than 37.5 and 37.5 cm3 or greater (P <0.05). No statistically significant difference in prostate-specific antigen, age, digital rectal examination, or Gleason score was found between each volumetric cutoff group. CONCLUSIONS: A significant percentage of men are diagnosed with CaP after a repeat biopsy. We have demonstrated that the percentage of CaP missed on the initial biopsy and detected on the repeat biopsy increases as the prostate volume increases. The results of our study suggest that in men with large prostates, traditional sextant biopsies may not be adequate to detect CaP. PMID- 12559291 TI - Quality of life after a diagnosis of prostate cancer among men of lower socioeconomic status: results from the Veterans Affairs Cancer of the Prostate Outcomes Study. AB - OBJECTIVES: To evaluate prospectively the health-related and disease-specific quality of life (QOL) at diagnosis and during the first year thereafter for patients with newly diagnosed prostate cancer who received care at Veterans Affairs Medical Centers. METHODS: Interviewers administered the European Organization for Research and Treatment of Cancer-QOL Questionnaire, a valid and reliable measure of health status, to 140 patients with prostate cancer at baseline (at diagnosis, before the initiation of treatment) and at 3 and 12 months thereafter at five Veterans Affairs Medical Centers. The mean changes from baseline values were analyzed statistically for patients with localized disease stratified by treatment group and separately for patients with metastatic disease. RESULTS: Among the 98 men with localized prostate cancer, significant disease-specific QOL changes noted at 3 and 12 months included worsening of urinary and sexual function among men treated with radical prostatectomy or radiotherapy and worsening of urinary function among those who opted for watchful waiting (each P <0.05). Among the 42 men with metastatic prostate cancer, significant decrements in role and social and sexual function were noted at 3 months, but had resolved on average by 12 months of follow-up. CONCLUSIONS: At 12 months, disease-specific QOL decrements persisted for patients with localized disease, but for patients with metastatic disease, disease-specific QOL appeared to return to near baseline (at diagnosis, before treatment initiation) function. Our study, among the first to assess the QOL at baseline before treatment, provides meaningful information on general treatment effects, which are directly relevant to clinicians when discussing treatment options with patients. PMID- 12559292 TI - Outcome, morbidity, and prognostic factors in post-prostatectomy radiotherapy: an Australian multicenter study. AB - OBJECTIVES: To report the outcome and morbidity data for patients treated with post-prostatectomy radiotherapy (PPRT) in a multicenter collaboration. METHODS: The case records of all patients treated with PPRT from 1996 to 1998 were reviewed. Survival was calculated using Kaplan-Meier methods. Potential prognostic factors were evaluated using the Cox proportional hazards regression model. Prostate-specific antigen (PSA) remission was defined as a PSA level of 0.2 ng/mL or less. Acute and late morbidities were documented. RESULTS: We reviewed the data of 115 patients, with a median follow-up from the start of PPRT of 28.7 months. Patients were treated with adjuvant intent (n = 23), for local recurrence (n = 27), or for a detectable PSA level (n = 65). The overall and cause-specific survival rate at 5 years was 73.7% and 81.4%, respectively. The biochemical disease-free survival rate was 69.6% at 2 years and 50% at 5 years. Factors predicting for subsequent relapse on multivariate analysis were pre-PPRT PSA (P = 0.013) and post-PPRT nadir (P <0.0001). Patients with a PSA greater than 1 ng/mL fared significantly worse than those with lower levels (P <0.0001). For patients with a pretreatment PSA of less than 1 ng/mL and an operative Gleason score of 7 or less, the 5-year projected biochemical disease-free survival rate was 71%. One case of grade 3 late proctitis was recorded, and 4 patients had continued grade 3 late urinary incontinence. CONCLUSIONS: Early use of PPRT is effective and well tolerated in patients at risk of, or with proven, local recurrence. PMID- 12559293 TI - Nocturnal penile tumescence and effects of complete spinal cord injury: possible physiologic mechanisms. AB - OBJECTIVES: To determine the role of the spinal cord in the initiation and/or modulation of nocturnal erectile activity, we measured nocturnal penile tumescence (NPT) in men with complete spinal cord injuries at known locations. METHODS: Eighteen men between the ages of 27 and 57 years (mean 45) with known complete spinal cord lesions and no history of other medical problems adversely affecting erectile function completed International Index of Erectile Function questionnaires and underwent RigiScan testing for two consecutive nights. RigiScan tracings were evaluated for the presence and quality of NPT activity. Erectile episodes lasting longer than 10 minutes with base and tip rigidity greater than 70% were designated "good"; the remainder were designated "borderline," "poor," or "absent." RESULTS: Of 9 men with cervical injuries and 9 with thoracic injuries, 8 and 3, respectively, had evidence of NPT. Of the 9 men with cervical injuries, 3 had one or more "good" nocturnal erections, and 1 of 9 men with thoracic injuries had one or more "good" nocturnal erections. The mean duration of the single longest erectile event was 26 minutes in men with cervical spinal injuries and 12 minutes in men with thoracic spinal injuries. Men with cervical and thoracic injuries had a mean overall International Index of Erectile Function score of 10 and 16 and a mean erectile domain score of 2 and 3, respectively. CONCLUSIONS: Spinal regulation is critical for nocturnal erectile activity, and the isolated thoracic cord is less effective than the isolated cervical cord in maintaining NPT. Despite the presence of nocturnal erectile activity, volitional erectile function in spinal cord-injured men is significantly impaired. PMID- 12559294 TI - How potent is potent? Evaluation of sexual function and bother in men who report potency after treatment for prostate cancer: data from CaPSURE. AB - OBJECTIVES: To characterize the association between potency and comprehensive sexual function. The accurate assessment of sexual function is critical for the evaluation of outcomes after treatment of prostate cancer. The assessments of potency typically used in this context, however, may be oversimplified. METHODS: CaPSURE is a large, observational database of men with prostate cancer. Participants complete health-related quality-of-life questionnaires, including the University of California, Los Angeles Prostate Cancer Index, every 6 months after treatment. A total of 5135 men completed at least one questionnaire and did not use medications for erectile function. The men were categorized as potent or impotent based on their ability to have erections and/or intercourse in the prior 4 weeks. Using the remaining questions on the Prostate Cancer Index, sexual function and bother scores were calculated for each group. RESULTS: Of the 5135 men, 27.4% were potent. The mean sexual function scores were 56 and 13 for potent and impotent men, respectively (P <0.0001). The corresponding mean bother scores were 62 and 36 (P <0.0001). The function scores ranged from 0 to 100 and 0 to 92 among potent and impotent men, respectively, and bother scores from 0 to 100 in both groups. Function was inversely associated with age in both groups, but bother did not change among potent men and ameliorated among impotent men. Individual Prostate Cancer Index questions correlated with potency to a variable extent. CONCLUSIONS: Although potent and impotent men have divergent sexual function and bother scores after treatment, the wide range of these scores in both groups denotes a complex picture of sexual function. The simple documentation of potency after treatment provides an insufficient measure of sexual health-related quality of life and should be supplemented with more comprehensive measures. PMID- 12559295 TI - Efficacy of sildenafil as adjuvant therapy to selective serotonin reuptake inhibitor in alleviating premature ejaculation. AB - OBJECTIVES: To evaluate the efficacy of sildenafil and selective serotonin reuptake inhibitor in alleviating premature ejaculation (PE) in patients in whom other treatments had failed. METHODS: Healthy men evaluated for primary PE graded their ejaculation on a scale of 0 to 8 (0 = almost never, 8 = almost always). The intravaginal ejaculatory latency time (IVELT) was graded on a scale of 0 to 3 (0 = longer than 5 minutes, 3 = shorter than 1 minute). The 138 men who scored their PE as 4 or greater and IVELT as 2 or greater comprised the study group. Psychological and behavioral counseling was provided during the study. PE was graded using the same scales 3 months after the initiation of each treatment. Topical 5% lidocaine ointment comprised the initial treatment: dissatisfied patients (PE grade 4 or greater, IVELT 2 or greater), took one tablet of paroxetine 20 mg for 30 days and then one tablet 7 hours before intercourse. Sildenafil was added to the treatment of patients dissatisfied with paroxetine alone. RESULTS: The mean initial PE grade was 5.67 +/- 0.13 and that for IVELT was 2.9 +/- 0.19 for all participants (mean age 28.7 years). Thirty-eight reported improvement (PE grade 2.0 +/- 0.8, P <0.01; IVELT 0.13 +/- 0.34, P <0.001) after local lidocaine application. Of the 100 treated with paroxetine, 42 reported improvement (PE grade 2.5 +/- 0.1, P <0.01; IVELT 0.28 +/- 0.46, P <0.001), and 56 of the remaining 58 who were treated with a combination of paroxetine and sildenafil reported improvement (PE grade 1.78 +/- 0.23, P <0.001; IVELT 0.16 +/- 0.37, P <0.001). Two patients remained dissatisfied with all treatment modalities. CONCLUSIONS: Sildenafil combined with paroxetine and psychological and behavioral counseling alleviated PE in patients in whom other treatments failed. PMID- 12559296 TI - Epidemiology of erectile dysfunction in four countries: cross-national study of the prevalence and correlates of erectile dysfunction. AB - OBJECTIVES: To measure the prevalence of erectile dysfunction (ED) in community based populations in Brazil, Italy, Japan, and Malaysia and to study its association with the demographic characteristics, medical conditions, and health related behavior. METHODS: In each country, a random sample of approximately 600 men aged 40 to 70 years was interviewed using a standardized questionnaire. All the data were self-reported. ED was assessed by the participants' "ability to attain and maintain an erection satisfactory for sexual intercourse," and the men were classified as not having ED if they answered "always" and as having mild, moderate, or complete ED if they answered "usually," "sometimes," or "never," respectively. RESULTS: The age-adjusted prevalence of moderate or complete ED was 34% in Japan, 22% in Malaysia, 17% in Italy, and 15% in Brazil. The overall age specific prevalence of moderate or complete ED was 9% for men aged 40 to 44 years, 12% for 45 to 49 years, 18% for 50 to 54 years, 29% for 55 to 59 years, 38% for 60 to 64 years, and 54% for those 65 to 70 years. The increased risk of ED was associated with diabetes, heart disease, lower urinary tract symptoms, heavy smoking, and depression and increased by 10% per year of age. It was inversely associated with education, physical activity, and alcohol drinking. CONCLUSIONS: ED is an international problem, the prevalence and severity of which increases with age. Despite national variations in prevalence, uniform associations were found between ED and medical conditions and lifestyle habits. PMID- 12559297 TI - Human sperm DNA integrity: correlation with sperm cytoplasmic droplets. AB - OBJECTIVES: To examine the retention of sperm cytoplasmic droplets (CD) and DNA denaturation (DD) in semen from fertile and infertile men. METHODS: Semen samples were obtained from consecutive nonazoospermic men presenting for infertility evaluation (n = 101) and fertile men presenting for vasectomy (n = 13). The standard semen parameters (sperm concentration, motility, and morphology), sperm DD, and sperm CD were monitored. Sperm DD was evaluated by flow cytometry analysis of acridine orange-treated spermatozoa and expressed as the percentage of spermatozoa demonstrating denatured DNA. RESULTS: The mean (+/-SE) percentages of spermatozoa with CD and DD were significantly higher in infertile than in fertile men (sperm CD 15.7% +/- 0.8% versus 4.8% +/- 0.7% and sperm DD 22.0% +/- 1.5% versus 10.8% +/- 1.8%, respectively). Sperm CD and DD were positively correlated (r = 0.59). Also, sperm CD and DD values correlated inversely with the standard semen parameters. CONCLUSIONS: Our data demonstrate that the retention of sperm CD correlates positively with sperm DD and that significantly higher sperm DD and CD are found in infertile than in fertile men. These data suggest that the enhanced susceptibility of sperm DNA to denaturation is associated with the abnormal disposal of residual sperm cytoplasm in the testis and/or epididymis. PMID- 12559298 TI - Does size and site matter for renal stones up to 30-mm in size in children treated by extracorporeal lithotripsy? AB - OBJECTIVES: To determine the safety and efficacy of extracorporeal shock wave lithotripsy (ESWL) in the treatment of pediatric nephrolithiasis and to analyze the impact of renal stone size and location on stone clearance. METHODS: This was a retrospective case series of pediatric patients (14 years or younger) undergoing lithotripsy using a Dornier MPL 9000 echo-guided lithotriptor. Patients were identified using an international coding and indexing system (9th edition) and ESWL registry. One hundred five patients were treated from January 1990 to December 2001. Stone clearance was assessed at 1 and 3 months. A stone free state was defined as no radiologic evidence of stone or fragments 3 mm or less. Treatment failure was analyzed to find any correlation with stone size and site. RESULTS: The overall stone-free rate was 95%. The treatment failed in 5 patients, who subsequently required ancillary procedures. Three patients received one session of ESWL, after which they were lost to treatment and were not analyzed further. Four of the five patients with treatment failure had a double-J stent; the stones were located in the lower pole calix in 2, renal pelvis and lower pole calix in 1, and at the ureteropelvic junction in 2 patients. The mean stone size in the treatment failure group was 15.88 mm compared with 14 mm in the stone-free group. The complication rate was 8%, but only 3 patients required admission. CONCLUSIONS: ESWL is highly effective for pediatric renal stones, with minimal morbidity. Stone clearance is not adversely affected by a stone size up to 30 mm; however, lower pole caliceal stones and stones impacted at the ureteropelvic junction have relatively poorer clearance. Eighty percent of patients who failed to clear their stones had a double-J stent. PMID- 12559301 TI - Onlay island flap urethroplasty: a comparative analysis of primary versus reoperative cases. AB - OBJECTIVES: To compare the outcome of the onlay technique in cases with an untouched urethral plate (group 1) and patients who had undergone previous hypospadias repairs (group 2). METHODS: The records of the 50 patients undergoing onlay island flap urethroplasty in the past 10 years were reviewed. The frequency of fistula formation in both groups was compared using the appropriate statistical methods. RESULTS: Seventeen and 33 patients were enrolled into groups 1 and 2, respectively. No statistically significant difference in patient age was present between the two groups (P >0.05). Flap necrosis in 2 patients and skin necrosis in 1 patient were encountered in group 2, with the meatus located at the penoscrotal region. Ischemic skin changes occurred in one primary patient with a mid-penile meatus. Urethrocutaneous fistula occurred in 2 (11.7%) of 17 primary cases and in 14 (42.4%) of 33 cases with multiple previous operations (Pearson chi-square test, P <0.05). A fistula rate of 61.5% was recorded in group 2 when the urethral meatus was located at the penoscrotal region. After withdrawal of the cases with penoscrotal meatus, the incidence of urethrocutaneous fistula was 6.7% (1 of 15) and 30% (6 of 20) in groups 1 and 2, respectively. CONCLUSIONS: Multiple previous operations affected the outcome of repair with the transverse onlay preputial flap, particularly in cases with a penoscrotal meatus. Repeat hypospadias repairs are more prone to complications. PMID- 12559303 TI - Histologic variants occupying a single prostatic tumor. PMID- 12559304 TI - Penile revascularization for treatment of erectile dysfunction secondary to blunt perineal trauma. PMID- 12559305 TI - Laparoscopic donor nephrectomy in a patient with previous upper pole partial nephrectomy. AB - A 56-year-old woman underwent laparoscopic partial nephrectomy for symptomatic angiomyolipoma. She then donated her remaining ipsilateral kidney for renal transplantation. The allograft had good, immediate function in the recipient. This is the first reported case of a patient undergoing renal donor nephrectomy after partial nephrectomy for angiomyolipoma. PMID- 12559306 TI - Management of priapism in the newborn. AB - We report a case of priapism in a newborn and the results of a review of the publications on neonatal priapism. A preterm newborn developed idiopathic priapism on the first day after birth. The priapism disappeared after 4 days of observation. Periods with normal penile erection were reported at follow-up after 20 months. The results of the literature review also advocate management by observation alone. PMID- 12559307 TI - Renal pseudoaneurysm after blunt renal trauma in a pediatric patient: management by angiographic embolization. AB - Renal pseudoaneurysm is a rare complication after blunt renal trauma. Only 18 cases have been reported in English-language published reports. We present a case of blunt renal trauma in an 11-year-old boy, complicated by delayed bleeding from a renal artery pseudoaneurysm. The patient was initially treated with conservative measures and was later treated with selective embolization of the pseudoaneurysm. The clinical presentation, management options, and clinical decisions are discussed. PMID- 12559308 TI - Ureteral obstruction and reconstruction in pelvic actinomycosis. AB - We present a case of ureteral obstruction secondary to pelvic actinomycosis. Despite stenting, prolonged antibiotic therapy, and debridement, the patient required ureteral resection and reconstruction. This condition may simulate advanced malignancy, and diagnostic suspicion lessens the need for radical extirpative surgery. PMID- 12559309 TI - Verrucous scrotal carcinoma in a patient with hypospadias: is there a possible association? AB - We report a case of a locally advanced verrucous scrotal cancer in a patient with hypospadias. Surgical excision was performed and no recurrence was noted with 14 months of follow-up. To our knowledge, this association has not been reported. Historically, scrotal carcinoma is occupation related, but none of the occupational predisposing factors was identified in this patient. This observation suggests that hypospadias and secondary chronic inflammation of the scrotal skin, caused by prolonged contact with urine, may contribute to an increased risk of verrucous carcinoma. PMID- 12559310 TI - Prone positioning for bulbar urethral reconstruction. AB - Exaggerated lithotomy is a surgical position associated with significant morbidity. We describe the use of the prone position for bulbar urethral reconstruction in a selected patient. We propose modified prone positioning as an alternative to exaggerated lithotomy for select patients undergoing reconstruction of the bulbar urethra. PMID- 12559311 TI - Penile and scrotal elephantiasis caused by indolent Chlamydia trachomatis infection. AB - Isolated penile and scrotal elephantiasis presents a diagnostic and therapeutic challenge. We report a case of chronic penile and scrotal elephantiasis caused by indolent Chlamydia trachomatis infection. The patient improved modestly with long term doxycycline. PMID- 12559312 TI - Giant multilocular cystadenoma of the prostate responsive to GnRH antagonists. AB - Giant multilocular cystadenoma of the prostate is a rare, benign, but locally recurrent, tumor that is usually treated by surgery. We report a case initially treated by surgical resection and followed by evaluation of serum prostate specific antigen values. After evidence of biochemical failure, pathologic recurrence was confirmed and treated by the gonadotropin-releasing hormone antagonist Lupron, with excellent results. The patient was stable without biochemical or radiologic evidence of progression during the last 2.5 years. This result offers a new treatment option for patients with this rare tumor. PMID- 12559313 TI - Multipoint methylation and expression analysis of tumor suppressor genes in human renal cancer cells. AB - OBJECTIVES: To analyze the methylation status and expression profiles of multiple tumor suppressor genes in renal cell carcinoma-derived cell lines. Aberrant promoter methylation is commonly found in human cancers. Nonetheless, it is challenging to demonstrate that methylation of a specific gene results in gene inactivation. METHODS: We simultaneously analyzed methylation and expression profiles of five putative tumor suppressor genes (p15, p16, Rb, BRCA1, and E cadherin) in 14 different cell lines using bisulfite genomic sequencing and reverse transcriptase-polymerase chain reaction. We also used multiplex polymerase chain reaction to identify homozygous deletions at the p15 and p16 loci. RESULTS: Expression of p16, BRCA1, and E-cadherin was maintained in 4 (29%) of 14 cell lines, regardless of the presence of methylation. Aberrant methylation of p16 was observed in 2 (14%), of BRCA1 in 1 (7%), and of E-cadherin in 9 (64%) of 14 cell lines. Concurrent methylation was observed among p16 and BRCA1 (1 [7%] of 14 cell lines) and among p16 and E-cadherin (1 [7%] of 14 cell lines). We detected homozygous deletion of p16 and p15 in 11 (78%) and 6 (43%) cell lines, respectively. CONCLUSIONS: The present data shows the presence of methylation does not always contribute to the loss of expression of tumor suppressor genes. Therefore, we must be cautious in interpreting the results of methylation assays- in particular, detection of methylation by nonquantitative methods. The data also demonstrated that multiple tumor suppressor genes are simultaneously inactivated in renal cell carcinoma-derived cell lines by distinctive mechanisms. PMID- 12559314 TI - Fiberoptic infrared spectroscopy: a novel tool for the analysis of urine and urinary salts in situ and in real time. AB - OBJECTIVES: To use infrared fiberoptic spectroscopy for the analysis of urinary salts in real time and with no sample processing; and to assess the practical role of this method for the quantitative measurement of the composition of urine and for the diagnosis of urolithiasis in patients. METHODS: Urine samples were obtained from two groups of patients: 24 patients with stone formation after shock wave lithotripsy and 24 normal subjects of similar age. Infrared absorption measurements were performed in real time, using infrared transmitting silver halide fibers. The absorption data were compared with the infrared absorption spectra of aqueous solutions prepared in our laboratory, with known concentrations of known urinary salts. The results were used for the study of the chemical composition of these salts in the urine samples and for a quantitative analysis of the concentration of the salts. RESULTS: We determined the composition of the stones in 20 of the 24 patients on the basis of the characteristic absorption peaks for the oxalates, carbonates, urates, and phosphates observed in their urinary samples. Using the method mentioned above, we found the concentration of different salts in urine with an average error of 20%. CONCLUSIONS: Fiberoptic infrared spectroscopy could be used as a new diagnostic tool for detecting different urinary salts in urine, finding their chemical composition, and determining their concentrations, without any sample preparation. PMID- 12559315 TI - Indomethacin-induced morphologic changes in the rat urinary bladder epithelium. AB - OBJECTIVES: To evaluate the morphologic changes in rat urothelium induced by indomethacin. Nonsteroidal anti-inflammatory drug-induced cystitis is a poorly recognized and under-reported condition. In addition to tiaprofenic acid, indomethacin has been reported to be associated with this condition. METHODS: Three groups were established: a control group (n = 10), a high-dose group (n = 10), treated with one intraperitoneal injection of indomethacin 20 mg/kg, and a therapeutic dose group (n = 10) in which oral indomethacin was administered 3.25 mg/kg body weight daily for 3 weeks. The animals were then killed and the bladders removed for light and electron microscopic studies. RESULTS: The light microscopic findings showed some focal epithelial degeneration that was more prominent in the high-dose group. When compared with the control group, both indomethacin groups revealed statistically increased numbers of mast cells in the mucosa (P <0.0001) and penetration of lanthanum nitrate through intercellular areas of the epithelium. Furthermore, the difference in mast cell counts between the high and therapeutic dose groups was also statistically significant (P <0.0001). CONCLUSIONS: Indomethacin resulted in histopathologic findings typical of interstitial cystitis, such as leaky bladder epithelium and mucosal mastocytosis. The true incidence of nonsteroidal anti-inflammatory drug-induced cystitis in humans must be clarified by prospective clinical trials. PMID- 12559316 TI - Role of uCyt+ in the detection and surveillance of urothelial carcinoma. AB - OBJECTIVES: To test the clinical value and role of uCyt+ as a noninvasive tool for the detection and surveillance of urothelial carcinoma. METHODS: Included in this prospective study were 235 patients (mean age 71.5 years, range 32 to 86). Of these, 98 patients had signs and symptoms suggestive of bladder cancer and 137 patients were being followed up after complete transurethral resection of superficial urothelial cancer (UC). All patients underwent urinary cytology and the uCyt+ test performed on ThinPrep (thin layer). All underwent subsequent cystoscopy and evaluation of any suspicious lesion by biopsy. RESULTS: A total of 102 patients had histologically proven UC. In the group of patients with signs and symptoms suspicious of UC, the sensitivity of cytology increased from 5% for G1 to 84.6% for G3 tumors; for uCyt+, it was 85% for G1, 100% for G2, and 92.3% for G3 tumors. Combining cytology and uCyt+, the sensitivity was 85% for G1 and 100% for G2 and G3. In the group of follow-up patients, the sensitivity of cytology increased from 4.3% for G1 to 94.4% for G3 tumors; for uCyt+, it was 78.2% for G1, 70% for G2, and 94.4% for G3 tumors. Combining both tests, the sensitivity was 78.2% for G1, 90% for G2, and 100% for G3. CONCLUSIONS: The uCyt+ is a valid test in the detection of UC of all grades and stages. It improves the sensitivity of cytology in low-grade tumors. The two tests combined may be a highly sensitive method to detect UC early in detection and surveillance. PMID- 12559317 TI - Glycoprotein A-80 in the human prostate: immunolocalization in prostatic intraepithelial neoplasia, carcinoma, radiation failure, and after neoadjuvant hormonal therapy. AB - OBJECTIVES: To review the expression of A-80 in prostate cancer and prostatic intraepithelial neoplasia and compare it with that of normal and hyperplastic prostatic tissue. The ability to recognize cancer after androgen deprivation therapy and residual and/or recurrent cancer after radiotherapy using A-80 staining was also examined. A-80 is a glycoprotein linked to exocrine differentiation that shows little or no expression in normal exocrine cells, but is selectively overexpressed in dysplasias and adenocarcinomas. METHODS: We studied 277 prostate samples with a monoclonal antibody (MAb) to A-80. We applied this MAb to paraffin sections of specimens of fetal (n = 12), benign prostatic hyperplasia (n = 26, from transurethral prostate resection specimens), atypical adenomatous hyperplasia (n = 11), prostate cancer (n = 103, from radical prostatectomy specimens), and autopsy (n = 7) tissue. In addition, 54 prostatectomy specimens after androgen-deprivation therapy and 64 specimens after radiotherapy were similarly studied. RESULTS: MAb A-80 stained the epithelial component of all 12 prostate specimens from fetal tissue; no staining was seen in normal adult prostatic tissue (0 of 7). In benign prostatic hyperplasia, sporadic cells reacted in 13% of cases (4 of 30); the atypical adenomatous hyperplasia samples were all negative (0 of 11). In patients with prostate cancer, more than 99% (102 of 103) stained positive, regardless of the grade or stage of cancer. Low and high-grade prostatic intraepithelial neoplasia reacted in 73% (38 of 52) and 92% (77 of 84) of cases, respectively. All 64 (100%) salvage prostatectomy samples after external beam radiotherapy stained positive for A-80. Carcinoma subsequent to neoadjuvant hormonal therapy stained positive in 98% (53 of 54) of cases. CONCLUSIONS: A-80 is useful in differentiating benign prostatic hyperplasia and atypical adenomatous hyperplasia from prostate cancer. Also, the strong A-80 reactions in most high-grade prostatic intraepithelial neoplasia provide strong molecular support to the precancerous nature of the lesion. In particular, A-80 staining of biopsies may be useful in detecting residual and/or recurrent prostate carcinoma after radiation or hormonal therapy. PMID- 12559318 TI - Robert H. Whitaker and the Whitaker test: a pressure-flow study of the upper urinary tract. PMID- 12559319 TI - Foley catheter removal techniques. PMID- 12559324 TI - Acute macular neuroretinopathy: a review of the literature. AB - Acute macular neuroretinopathy (AMNR) is a rare condition that produces transient or permanent visual impairment. Typical cases have acute onset multifocal scotomas that correspond rather precisely with reddish, flat, or depressed circumscribed lesions in the macula. These lesions are wedge-shaped and generally point toward the fovea. The pathophysiology of AMNR is unclear, the causes are uncertain, and there is no specific treatment for this condition. This review summarizes the presentation, possible risk factors, and prognosis of the 41 cases of AMNR reported in the published, English-language literature from 1975 through April 2002. Possible areas for future research into the etiology of this rare condition are discussed. PMID- 12559325 TI - Paraneoplastic retinopathies and optic neuropathies. AB - Unusual neuro-ophthalmologic symptoms and signs that go unexplained should warrant a thorough investigation for paraneoplastic syndromes. Although these syndromes are rare, these clinical manifestations can herald an unsuspected, underlying malignancy that could be treated early and aggressively. This point underscores the importance of distinguishing and understanding the various, sometimes subtle, presentations of ocular paraneoplastic syndromes. Outlined in this review article are diagnostic features useful in differentiating cancer associated retinopathy, melanoma-associated retinopathy, and paraneoplastic optic neuropathy. These must also be distinguished from non-cancer-related eye disorders that may clinically resemble cancer-associated retinopathy. The associated antibodies and histopathology of each syndrome are presented to help in the understanding of these autoimmune phenomena. Treatment outcomes from reported cases are summarized, and some potential novel immunotherapies are also discussed. PMID- 12559326 TI - Rhabdomyosarcoma: review for the ophthalmologist. AB - Rhabdomyosarcoma is a malignant neoplasm that is composed of cells with histopathologic features of striated muscle in various stages of embryogenesis. It can occur in several sites in the body, including the ocular region. Ocular rhabdomyosarcoma is defined as the occurrence of this tumor in the area of the eye. Most ocular rhabdomyosarcomas arise in the soft tissues of the orbit but they can rarely occur in the other ocular adnexal structures and even within the eye. The purpose of this review is to provide a brief overview of rhabdomyosarcoma and a more detailed review of orbital rhabdomyosarcoma, with emphasis on changing concepts in the diagnosis and management of this ophthalmic neoplasm. PMID- 12559328 TI - The evolution of endonasal dacryocystorhinostomy. AB - Dacryocystorhinostomy is an important treatment in the relief of tearing. Through the years, there have been several advances in this procedure. Once performed only from an external approach, the advent of rigid endoscopes and endoscopic instrumentation has made the endonasal approach a reality. Advantages of the endonasal approach include lack of a cutaneous incision and excellent visualization of intranasal pathology, which is often the cause of dacryocystorhinostomy failure. Preoperative evaluation including a detailed medical history, physical examination with office endoscopy, and imaging, as well as postoperative care are important. Surgical technique with detailed knowledge of intranasal anatomy and meticulous attention to hemostasis are critical. Endonasal laser-assisted dacryocystorhinostomy is also performed today, and special preoperative considerations and key features of the lasers available are important in the selection of a laser for tissue or bone ablation during the procedure. Endoscopic conjunctivodacryocystorhinostomy is performed today, and given the critical nature of proper length and placement of the Jones tube intranasally, provides the significant advantage of intranasal visualization. Endoscopic dacryocystorhinostomy in children is also performed today; however, challenges such as small nasal anatomy and maintenance of the intranasal osteomy postoperatively in a child are significant issues. PMID- 12559327 TI - Complex visual hallucinations in the visually impaired: the Charles Bonnet Syndrome. AB - Visually impaired patients may experience complex visual hallucinations, a condition known as the Charles Bonnet Syndrome. Patients usually possess insight into the unreality of their visual experiences, which are commonly pleasant but may sometimes cause distress. The hallucinations consist of well-defined, organized, and clear images over which the subject has little control. It is believed that they represent release phenomena due to de-afferentation of the visual association areas of the cerebral cortex, leading to a form of phantom vision. Cognitive defects, social isolation, and sensory deprivation have also been implicated in the etiology of this condition. This condition, which is most common in the elderly, frequently goes unrecognized in clinical practice, due to both lack of awareness among doctors and patients' reluctance to admit to hallucinatory experiences, for fear of being labeled mentally unstable. Furthermore, patients who comprehend the unreality of their hallucinations may be distressed by the real fear of imminent insanity. Sensitive and sympathetic history taking is essential to ascertain the existence of hallucinations. Reassurance and explanation that the visions are benign and do not signify mental illness have a powerful therapeutic effect. Hallucinatory activity may terminate spontaneously, on improving visual function or on addressing social isolation. There is no universally effective drug treatment but anticonvulsants may play a limited role in aborting the hallucinations. Physician awareness and empathy are the cornerstones of management. PMID- 12559329 TI - The double vision decision. AB - A 50-year-old man experienced the acute onset of ophthalmoplegia, ataxia, and hyporeflexia. Evaluation led to the diagnosis of Miller Fisher syndrome (MFS). Appropriate evaluation and management of MFS is discussed. PMID- 12559330 TI - Motion perception in glaucoma patients: a review. AB - Most of the histopathological and psychophysical studies in glaucoma reveal a preferential damage to the magnocellular (M) pathway although a few of them support a damage to the parvocellular (P) pathway as well. In glaucoma, the visual fields are usually evaluated by conventional perimetry. However, it has been demonstrated that 20-40% of ganglion cells are lost before field defects are detected using conventional perimetry. Therefore, new psychophysical tests have recently been designed in order to specifically isolate and evaluate the visual mechanisms that are impaired at the early stages of glaucoma. In this context, several authors have addressed the issue of motion perception under the hypothesis of a predominant damage of the M pathway in glaucoma, and that motion perception is mediated mainly by M pathway. The results of these studies depict a large variation in the percentage of patients showing anomalous motion perception. Overall, motion thresholds are elevated in both glaucoma and ocular hypertensive patients as compared to control subjects, irrespective of the stimulus size and eccentricity. The test which discriminates best between patients and normal subjects is motion perimetry. The visual field defects in glaucoma patients identified by conventional perimetry and motion perimetry are similar, but the sizes of the defects are usually larger with motion perimetry. However, motion tests in central vision have no correlation with visual field defect on conventional perimetry. In glaucoma, loss of performance on motion perception tests does not necessarily support the existence of a specific deficit in the M pathway, because some behavioral studies suggest that the P pathway can also mediate motion perception. It is also difficult to conclude that motion perception is specifically affected in glaucoma because most of these studies do not yield a comparison with other visual functions. Despite these difficulties, localized motion perception tests at eccentricities of more than 15 degrees can be considered as a promising diagnostic tool. PMID- 12559331 TI - Ocular manifestations in the inherited DNA repair disorders. AB - Deoxyribonucleic acid (DNA) repair is a fundamental process designed to keep the integrity of genomic DNA that is continuously challenged by intrinsic or environmental induced alterations. Numerous genes involved in DNA repair have been cloned and are involved in different DNA repair pathways: base excision repair, nucleotide excision repair, mismatch repair, DNA recombination. Inherited conditions due to mutations in DNA repair genes include mainly: xeroderma pigmentosum, Cockayne syndrome, Trichothiodystrophy, Bloom syndrome, Rothmund Thomson syndrome, and Werner syndrome. Minor to major ocular manifestations occur in these syndromes. For example, eyelid skin cancers in xeroderma pigmentosum and retinal dystrophy in Cockayne syndrome are major features of these syndromes. This review focuses on the DNA repair pathways, the general and ocular features of the related syndromes, the laboratory tests useful for diagnosis, and the general processes implied with DNA repair (ultraviolet sensitivity, carcinogenesis, apoptosis, oxydative stress, and premature aging). PMID- 12559333 TI - Rupture of the aorta following road traffic accidents in the United Kingdom 1992 1999. The results of the co-operative crash injury study. AB - OBJECTIVE: The true incidence and survivability of blunt traumatic aortic rupture following road traffic accidents in the UK is unclear. The objective of this study was to determine the extent of blunt traumatic aortic rupture in the UK after road traffic accidents and the conditions under which it occurs. METHODS: Data for the study was obtained from the Co-operative Crash Injury Study database. Road traffic accidents that happened between 1992 and 1999 and included in the Co-operative Crash Injury Study database were retrospectively investigated. RESULTS: A total of 8285 vehicles carrying 14,435 occupants were involved in 7067 accidents. There were 132 cases of blunt traumatic aortic rupture, of which the scene survival was 9% and the overall mortality was 98%. Twenty-one percent of all fatalities had blunt traumatic aortic rupture (130/613). Twenty-nine percent were due to frontal impacts and 44% were due to side impacts. Twelve percent of the blunt traumatic aortic rupture cases in frontal vehicle impacts were wearing seat belts and had airbag protection and 19% had no restraint mechanism. The Equivalent Test Speed of the accident vehicles, (where equivalent test speed provides an estimate of the vehicle impact severity and not an estimate of the vehicle speed at the time of the accident), ranged from 30 to 110 km/h in frontal impacts and from 15 to 82 km/h in side impacts. CONCLUSION: Blunt traumatic aortic rupture carries a high mortality and occurred in 21% of car occupant deaths in this sample of road traffic accidents. Impact scenarios varied but were most common from the side. The use of an airbag or seat belt does not eliminate risk. The injury can occur at low severity impacts particularly in side impact. PMID- 12559335 TI - Off-pump apicoaortic conduit insertion for high-risk patients with aortic stenosis. AB - OBJECTIVES: Recently, we implanted an apicoaortic conduit off-pump in three high risk patients with severe aortic stenosis. METHODS: A muscle-coring device was utilized to create the apical outflow tract followed by insertion of a rigid apical connector. A valved conduit was then connected to the descending thoracic aorta and to the apical connector graft. RESULTS: A stentless porcine bioprosthesis was implanted in two patients and a stented valve in one. The left ventricular (LV)-aortic gradients were reduced from a mean of 66 to 28 mmHg. CONCLUSIONS: With modification of an existing technique, apicoaortic conduit insertion can be performed safely off-pump. This technique can be applied to complex forms of LV outflow obstruction and high-risk patients. PMID- 12559334 TI - Aortic arch reconstruction using regional perfusion without circulatory arrest. AB - OBJECTIVES: Deep hypothermic circulatory arrest during repair of aortic arch anomalies may induce neurological complications or myocardial injury. Regional cerebral and myocardial perfusion may eliminate those potential side effects. METHODS: From March 2000 to March 2002, 48 neonates or infants with complex arch anomaly were operated on using the regional perfusion technique. Thirty-three patients were male and the median age was 24 days (range 5-301 days). Preoperative diagnosis consisted of coarctation or interruption of the aorta associated with ventricular septal defect (group I, n = 26) and arch anomaly with complex intracardiac defects such as hypoplastic left heart syndrome or its variants (group II, n = 22). Arterial cannula was inserted through the innominate artery and the flow rate was regulated to about 50-100 ml/kg per min during regional perfusion. Simultaneous myocardial perfusion was maintained using a Y connected infusion line. Cardioplegia was applied during intracardiac repair. RESULTS: Cardiopulmonary bypass and aortic cross-clamp times were 154 +/- 49 and 39 +/- 34 min, respectively. Temporary circulatory arrest for intracardiac procedures was performed in eight patients. However, the mean arrest time was minimized (range 1-18 min). The descending aorta clamping time was 33 +/- 16 min. Operative mortality rates in each group were 0 and 18.2% (0/26 and 4/22). Late mortality rates were 0 and 11.1% (0/26 and 2/18) during 9.1 months of follow-up. Complications consisted of low cardiac output in eight cases, transient neurological problems in two cases, and transient renal insufficiency in two cases, respectively. CONCLUSIONS: Regional perfusion is feasible and can be used with acceptable results. It may reduce potential complications following aortic arch reconstruction using circulatory arrest. However, repair of aortic arch in the patients with complex intracardiac defects still imposes a significant rate of mortality and morbidity. PMID- 12559336 TI - Reduction of neurologic complications by intra-aortic filtration in patients undergoing combined intracardiac and CABG procedures. AB - OBJECTIVE: Patients undergoing combined intracardiac and coronary artery bypass grafting (CABG) procedures are at high risk for neurologic events. Previously the Multicenter Study of Perioperative Ischemia (McSPI) Study Group demonstrated that risk factors for neurologic complications in these patients related primarily to embolization. OBJECTIVE: This study investigated risk factors and outcomes in 185 high-risk patients receiving intra-aortic filtration during combined intracardiac and CABG procedures. METHODS: Seventeen European centers comprising the International Council of Emboli Management (ICEM) pooled data on patients receiving intra-aortic filtration (EMBOL-X, Mountain View, CA, USA) into a consecutive enrollment registry. Preoperative, intraoperative, and outcome variables were compared between ICEM patients undergoing combined CABG and intracardiac procedures and the corresponding McSPI subgroup. RESULTS: Type I outcomes (stroke, transient ischemic attack, coma, death due to neurologic causes) occurred in 2.2% (4/185) of ICEM patients compared to 8.4% (23/273) of McSPI patients. No ICEM patient suffered a fatal stroke compared with five in the McSPI study (1.8%). One nonfatal stroke was observed in the ICEM group (0.5%) and 16 were observed in the McSPI group (5.9%). Three patients (1.6%) were reported with transient ischemic attack in the ICEM group, compared to two in the McSPI study (0.7%). One of the reported patients in the ICEM group with transient ischemic attack also suffered a coma that was still evident at discharge. In all, four patients suffered Type I neurologic events in the ICEM group (2.2%), compared to 23 in the McSPI group (8.4%). In the ICEM study group, four patients (2.2%) died due to non-neurologic causes, compared to 18 in the McSPI group (6.6%). CONCLUSIONS: Even with a significantly worse preoperative profile, high risk patients receiving intra-aortic filtration suffered Type I cerebral injury 74% less often than patients in the McSPI group. PMID- 12559337 TI - Vasoplegic syndrome after off-pump coronary artery bypass surgery. AB - OBJECTIVE: The vasoplegic syndrome (VS) has been implicated in life-threatening complications after open heart surgery, where the whole-body inflammatory reaction is attributed to the cardiopulmonary bypass (CPB). Off-pump coronary artery bypass grafting (OPCAB) has been recently achieving growing enthusiasm mainly due avoiding the side effects of CPB. However herein the occurrence of VS in OPCAB is reported. METHODS: The vasoplegic syndrome usual findings occurring in the early postoperative period include severe hypotension, tachycardia, normal or elevated cardiac output and low systemic vascular resistance. Four patients underwent to OPCAB presented all the signs of VS intraoperatively or within the first 6 postoperative h. RESULTS: The patients needed aggressive vasoactive drug support for hemodynamic stabilization and all of them developed complications. These patients also had tendency to require administration of blood and blood derivatives due to diffuse and oozing type bleeding. Mean intensive care unit stay of surviving patients was 70 h and mean period of postoperative hospitalization was 9 days. Tumor necrosis factor-alpha blood levels in one patient were elevated postoperatively though no signs of infection were observed. One patient died. CONCLUSIONS: Although vasoplegic syndrome can complicate OPCAB surgery, the rationale for avoiding CPB remains valid considering the benefits provided by OPCAB. PMID- 12559338 TI - Off-pump coronary artery bypass surgery does not reduce gastrointestinal complications. AB - OBJECTIVES: Gastrointestinal (GI) complications following on-pump coronary artery bypass grafting (CABG) are rare, but carry a high mortality rate. Prolonged cardiopulmonary bypass (CPB) has been associated with a higher incidence of such complications. Little is known about the effect of avoiding CPB on GI complications. Our hypothesis was that off-pump CABG might reduce such complications. METHODS: A total of 2327 consecutive cases undergoing isolated CABG between April 1997 and May 2001 were identified from four consultants' practice at the two cardiothoracic centres involved in this study. We performed a multivariable logistic regression analysis to identify the risk factors for development of post-operative GI complications. Potential risk factors considered in the logistic model were age, sex, angina, ejection fraction, peripheral vascular disease, renal dysfunction, redo operations, previous GI complications, priority of surgery and the use of CPB. RESULTS: A total of 1210 cases were performed on CPB, compared to 1117 off-pump. The incidence of GI complications was 1.2% (n = 14) in the on-pump group and 1.6% (n = 18) in the off-pump group (P = 0.347). The incidence of in-hospital mortality, in the patients who had a GI complication, was 28.6% (n = 4) and 22.2% (n = 4), respectively (P = 0.681). The results of the logistic regression analysis showed that renal dysfunction, advancing age and previous history of GI surgery are significant risk factors for GI complications after coronary bypass surgery whether CPB is used or not. CONCLUSIONS: Our study suggests that off-pump and on-pump techniques are similar in the rates of GI complications. We suggest that a properly designed randomized control trial is needed to verify our findings. PMID- 12559339 TI - Effects of atrial fibrillation on coronary artery bypass graft flow. AB - OBJECTIVES: No detailed studies exist of coronary artery bypass graft flow during atrial fibrillation. We examined the effects on bypass graft flow of atrial fibrillation following coronary artery bypass grafting. METHODS: Immediately after surgical revisualization, atrial fibrillation was induced in 18 patients by high frequency atrial pacing. Hemodynamic variables were measured in sinus rhythm and atrial fibrillation. The graft flow in pedicled left internal thoracic artery grafts and in saphenous vein grafts was also measured using transit-time flowmetry. RESULTS: Left internal thoracic artery graft flow had a greater diastolic component than saphenous vein graft flow, as shown by the percent diastolic time-flow integral (86 +/- 10% in the left thoracic artery and 62 +/- 12% in the saphenous vein, P < 0.0001). The induced atrial fibrillation caused significant deterioration in hemodynamics: heart rate and central venous pressure increased, and mean arterial pressure and cardiac index decreased (all P < 0.0025). In left internal thoracic artery grafts (n = 18) and also in saphenous vein grafts (n = 20), graft flow decreased significantly with atrial fibrillation (44.3 +/- 26.2 to 26.2 +/- 20.7 ml/min in the left internal thoracic artery, P = 0.0003; 39.7 +/- 15.6 to 33.3 +/- 14.3 ml/min in the saphenous vein, P = 0.001). The reduction in graft flow due to atrial fibrillation was much larger in left internal thoracic artery grafts than in saphenous vein grafts (P = 0.0008). CONCLUSIONS: Direct measurement of coronary artery bypass graft flow shows that atrial fibrillation after surgery significantly reduces graft flow. The effect is much larger in left internal thoracic artery grafts with their strong diastolic component than in saphenous vein grafts. PMID- 12559340 TI - Coronary artery bypass grafting (CABG) after initially successful percutaneous transluminal coronary angioplasty (PTCA): a review of 17 years experience. AB - OBJECTIVE: Patients who undergo successful percutaneous transluminal coronary angioplasty (PTCA) may subsequently require operative myocardial revascularization. This review examines whether prior successful PTCA alters outcomes following subsequent coronary artery bypass grafting (CABG). The costs of interventional cardiology procedures and definitive surgery were also examined. METHODS: From January 1981 through December 1997, 361 patients underwent CABG following initially successful PTCA (interval group). This group was compared with 11,909 patients who underwent CABG as the primary intervention for coronary artery disease (control group). RESULTS: The average time interval to CABG following initial PTCA was 13.7 months. The post-CABG myocardial infarction rate was 4% for patients in the interval group and 3% for patients in the control group. The 30-day mortality was similar for both patient groups (2%). For the interval group, the average cost of total interventional management was 24,220 dollars per patient. This included average costs of 13,873 dollars for CABG and 10,347 dollars for all preoperative interventional cardiology procedures. CONCLUSION: There is little doubt that PTCA procedures may provide successful myocardial revascularization. However, these procedures often need to be repeated over time and may serve only to delay coronary surgery, at substantial financial and personal cost. PMID- 12559341 TI - Surgical treatment of complete A-V canal defects in children before 3 months of age. AB - OBJECTIVES: Surgical repair of complete A-V canal defects (CAVCD) is a well established procedure which is currently performed in infancy. The aim of this study is to evaluate surgical results of correction in early infancy in comparison to older age. METHODS: From January 1985 to March 2001, 119 consecutive patients (age range 27 days to 83 months, mean 6.7 months) underwent repair of CAVCD in our Institution. Forms with unbalanced ventricles in association with Fallot's tetralogy or heterotaxia were excluded from this series. Fifty-eight patients (49%) underwent correction before 3 months of age (Group A), and 61 patients (51%) after 3 months (Group B). Surgical repair was accomplished with a double patch technique in 100 patients (84%). Associated surgical lesions were treated simultaneously in 48 patients (40%). RESULTS: There were 11 operative deaths (<30 days) (two in Group A (3.4%) and nine in Group B (15%)) (P = 0.05). The remaining patients were discharged home in good haemodynamic condition. Reoperation for postoperative left A-V incompetence occurred in five patients in Group A and in eight patients in Group B. There were eight late deaths (three in Group A and five in Group B), of which four were non cardiac related. At a mean follow-up time of 80 months (range 2-184 months) 100 patients are asymptomatic and well, and free from oral medication. Echocardiographic examination showed absent or mild residual left A-V valve incompetence in 91 patients (49 in Group A and 42 in Group B) and moderate left A V valve incompetence in nine patients (four in Group A and five in Group B). Kaplan-Meier survival estimates at 10 years were 90% for Group A and 75% for Group B. Kaplan-Meier freedom from reoperation at 10 years was 89% for Group A and 84% for Group B. CONCLUSIONS: Our data demonstrate that repair of CAVCD under 3 months of age is the ideal approach to this malformation with a lower mortality rate at operation compared to older patients. Logistic analysis showed that an operative age >3 months is, compared to an age < or =3 months, an incremental risk factor for hospital mortality with an odds ratio of 4.8 (95% confidence limit 1-23.5) (P = 0.05). In the long term, freedom from reoperation for left A-V valve incompetence is higher when compared to children repaired at an older age. PMID- 12559342 TI - Detection of telomerase activity in bronchial lavage as an adjunct to cytological diagnosis in lung cancer. AB - OBJECTIVE: Definitive diagnosis of lung cancer with conventional methods may sometimes be difficult in clinical practice. Telomerase is a ribonucleoprotein DNA polymerase that maintains the telomeric region of chromosomes during successive rounds of cell division. Telomerase activity in body cavity fluids has been advocated to be a potential diagnostic marker for malignancy. We investigated the diagnostic value of telomerase activity in bronchial lavage samples of patients undergoing diagnosis of lung cancer. METHODS: A total of 29 bronchial lavage samples were collected from patients in whom the diagnosis was confirmed with cytological and/or histological examinations. Patients were classified as lung cancer patients (Group 1, n = 22) and patients with benign disease (Group 2, n = 7). Telomerase activity was determined with polymerase chain reaction-based TRAP (The telomeric repeat amplification protocol) assay. RESULTS: Cytological examination was diagnostic in 12 (54.5%) of 22 patients in Group 1, and in all seven patients of Group 2 (P = 0.063). Telomerase activity was positive in 16 (72.7%) of Group 1 patients, while it was positive in only 1 (14.3%) sample of a lung abscess in Group 2 (P = 0.011). The sensitivity rate of cytological examination when combined with telomerase activity (81.8%) was significantly greater than that of cytological examination alone (54.5%) (P = 0.031). The sensitivity and specificity of telomerase activity were 72.7 and 85.7%, respectively. Telomerase activity had a positive predictive value as 0.94 and negative predictive value as 0.50. Diagnostic accuracy of telomerase activity was 75.8%. CONCLUSION: Telomerase activity in bronchial lavage is a highly sensitive diagnostic biomarker for malignancy and a potential complementary diagnostic technique to cytological examination in the diagnosis of lung cancer. PMID- 12559343 TI - Predicting pulmonary complications after pneumonectomy for lung cancer. AB - OBJECTIVES: Patients undergoing pneumonectomy for lung cancer are thought to be at high risk for the development of postoperative pulmonary complications (PC) and these complications are associated with high mortality rates. The purpose of this study was to identify independent factors associated with increased risk for the development of postoperative PC after pneumonectomy for lung cancer, and to assess the usefulness of predicted pulmonary function to identify high risk patients and other adverse outcomes. PATIENTS AND METHODS: We reviewed retrospectively 242 patients undergoing pneumonectomy for lung cancer during a 12 year period. Perioperative data (clinical, pulmonary function test, and surgical) were recorded to identify risk factors of PC by univariate and multivariate analyses. RESULTS: Overall mortality and morbidity rates were 5.4 and 59%, respectively. Thirty-four patients (14%) developed PC (acute respiratory failure, ARF = 8.7%, reintubation = 5.4%, pneumonia = 3.3%, atelectasis = 2.9%, postpneumonectomy pulmonary edema = 2.5%, mechanical ventilation more than 24 h = 1.2%, pneumothorax = 0.8%). Patients with surgical (P < 0.001), cardiac (P < 0.001) and other complications (P < 0.01) had higher incidence of PC than those without postoperative complications. Intensive care unit stay (53 +/- 39 h vs. 35 +/- 19 h; P < 0.001) and hospital stay (18 +/- 11 days vs. 12 +/- 7 days; P < 0.001) was significantly longer in patients with PC. The mortality rate associated with PC was 35.5% (P < 0.001). By univariate analysis, it was found that older patients (P = 0.007), chronic obstructive pulmonary disease (COPD) (P = 0.023), heart disease (P = 0.019), no previous record of chest physiotherapy (P = 0.008), poor predicted postoperative forced expiratory volume in 1s (ppo-FEV1) (P = 0.001), and prolonged anesthetic time (P < 0.001) were related with higher risk of PC. In the multiple logistic regression model, the anesthetic time (minutes; odds ratio, OR = 1.012), ppo-FEV1 (ml/s; OR = 0.998), heart disease (OR = 2.703), no previous record of previous chest physiotherapy (OR = 2.639), and COPD (OR = 2.277) were independent risk factors of PC. CONCLUSIONS: PC after pneumonectomy are associated with high mortality rates. Careful attention must be paid to patients with COPD and heart disease. Our results confirm the relevance of previous chest physiotherapy and the importance of the length of the surgical procedure to minimize the incidence of PC. The predicted pulmonary function (ppo FEV1) may be useful to identify high risk patients for PC development and adverse outcomes. PMID- 12559344 TI - Evaluations of bronchoplasty and pulmonary artery reconstruction for bronchogenic carcinoma. AB - OBJECTIVE: To evaluate the surgical results of bronchovascular reconstruction and the prognostic factors for lung cancer. METHODS: From 1976 to 1995, 78 patients with a mean age of 55.1 years (range 26-69 years) underwent bronchoplasty for non small-cell lung cancer (NSCLC) including pulmonary artery (PA) reconstruction in 21 patients. There were 47 right upper lobectomies (60.3%), 24 left upper lobectomies (30.8%), and seven other atypical types of operations (8.9%). The bronchoplasty was a full sleeve in 71 patients, and a bronchial wedge resection in seven. Thirteen PA tangential resections and eight PA sleeve resections were performed. Tissue diagnosis was squamous cell carcinoma in 56 patients, adenocarcinoma in six, adenosquamous carcinoma in ten, neuroendocrine carcinoma in two and others in four. No patient had a microscopically positive bronchial resection margin. The follow up is complete for all patients. Seventy-five patients were statistically analyzed using STATA software. The survival rate was calculated with life table method. Comparisons of the difference of survival rates between groups were made according to the log-rank test. RESULTS: The operative mortality rate (30 days) was 3.8% (3/78). The prolonged atelectasis necessitating repeated bronchoscopy was the most common major complication which occurred in 12 patients (16%). Tumor recurrence around the anastomotic site confirmed by bronchoscopic biopsy was observed in four patients. The overall survival at 5 and 10 years was 48.9 and 38.8%, respectively. The 5- and 10-year survival for patients with stage I disease were 66.1 and 57.5%, and for patients with stage II were 62.8 and 44.2%, respectively. The 3- and 5-year survivals for patients with stage III were 11.1 and 0%, respectively (P = 0.0000). The 5-year survival rates for those with N0 tumor (n = 36) were 63.3%, 53.6% for those with N1 (n = 26), and with no survivors for N2 (n = 13), respectively (P = 0.0000). The 5- and 10- year survival rates with bronchoplasty (n = 54) were 55.0 and 47.8%, and 33.3 and 16.7% with bronchovascular reconstruction (n = 21), respectively (P = 0.0033). Multivariate analysis showed that long-term results were influenced chiefly by nodal stage among five factors of pT, pN, bronchoplasty with or without PA reconstruction, cell types, and postoperative adjuvants (P = 0.004). CONCLUSIONS: Any type of lobectomy with bronchial reconstruction is an adequate cancer operation for both compromised and uncompromised patients especially in patients with stages I and II lung cancer with reasonably good results. Sleeve lobectomy with PA reconstruction may finally be indicated in patients considered compromised because of cardiac or respiratory impairment contraindicating pneumonectomy. PMID- 12559345 TI - Intraoperative radioguided sentinel lymph node biopsy in non-small cell lung cancer. AB - OBJECTIVES: The aim of this study was to determine the accuracy and the role of the sentinel lymph node (SLN) technique in patients with early non-small cell lung cancer (NSCLC). METHODS: This study was carried out on 29 consecutive patients (M/F = 24:5, mean age 65.9 +/- 7.1 years) with resectable NSCLC (Stage IA-IB). Intraoperative injection with a (99m)Tc-nanocolloid suspension was performed in the first ten patients; the following patients were injected under computed tomography scan guidance. A total dose of 37 MBq (1 ml) was administered in two to four divided aliquots (depending on the size), injected in the periphery of the tumour. Intraoperative radioactivity counting started a mean of 1 h (range 50-70 min) after the injection. The SLN was defined as the node with the highest count rate using a handheld gamma probe counter. Resection with mediastinal node dissection was performed and findings were correlated with histologic and immunohistochemistry (IHC) examination. RESULTS: Three of the 29 patients did not have NSCLC (two benign lesions, and one metastatic breast tumour) and were excluded. The SLN was identified in 25/26 (96.1%) patients (a total of 31 SLNs); 7/31 (22.5%) of the SLNs were positive for metastatic involvement after histologic and IHC examination. One inaccurately identified SLN was encountered (3.8%). CONCLUSIONS: These preliminary results demonstrate the feasibility of this procedure in identifying the first site of potential nodal metastases of NSCLC. The actual clinical impact of this procedure remains to be elucidated by further investigation in larger groups of patients. PMID- 12559346 TI - Prognostic prediction of the immunohistochemical expression of p53 and p16 in resected non-small cell lung cancer. AB - OBJECTIVE: p53 and p16(INK4) are the common and important tumor suppressor genes. Aberrant expression of p53 or p16 protein has been reported in various malignancies including lung cancer. Our aim was to investigate the association of p53 and p16 expression in resected non-small cell lung carcinoma (NSCLC) and evaluated their correlation with clinocopathologic features and survival. METHODS: p16 and p53 expression were detected by immunohistochemical analysis of 90 paraffin specimens of resected NSCLC, including 35 squamous cell carcinoma, 47 adenocarcinoma, and eight large cell carcinoma, between stages I and IV. The immunohistochemical study was performed using the labeled streptavidine-biotin method with anti-p53 and anti-p16 monoclonal antibodies. RESULTS: Fifty-two (57.8%) and 36 (40%) of 90 patients revealed aberrant immunostaining for p53 (p53+) and p16 (p16+), respectively. While 19 cases (21.1%) showed abnormal immunoreactivity for both p16 and p53. (p53+/p16+). There was no correlation of p53 or p16 expression with the clinicopathologic features. The Kaplan-Meier survival analysis demonstrated that patients with p16+, p53+, late stages, and nodal or distal metastasis had poor survival status (P = 0.006, 0.013, <0.001, <0.001 and 0.018, respectively). Further analysis demonstrated that p53 status was a significant prognostic factor in stage I NSCLCs (P < 0.001), and p16 status in stage I and II NSCLCs (P < 0.001, P = 0.003, respectively). Furthermore, patients whose tumors were both p53 and p16 aberrant expression had worse outcome compared with those whose tumors were both normal expression of p53 and p16 (5 year survival rate: 5 vs. 76%, P < 0.001). In Cox's regression model, the aberrant expression of p16, p53, advanced stages and combined aberrant expression of p53/p16 survived for a significant shorter period. CONCLUSIONS: The results indicated that aberrant expression of p16 and p53 are significant and independent, predictable prognostic factors for resected NSCLC, especially in early stage of NSCLCs. The worst prognosis was seen in patients whose tumors had both aberrant expression of p53 and p16. Further prospective trials may be aimed at confirming and validating these results. PMID- 12559347 TI - Unsuspected residual disease at the resection margin after surgery for lung cancer: fate of patients after long-term follow-up. AB - OBJECTIVE: This retrospective study evaluates the survival impact of the residual margin disease after bronchial resection for cancer and suggests tactics in cases of microresidual disease. METHODS: Between March 1988 and 1998, 4530 consecutive patients underwent surgery for non-small cell lung cancer at our institution. Only incomplete resections after microscopic evaluation (R1) were included in the study. Residual tumour cells were found on the bronchial resection margins of 39 lobectomies, 12 pneumonectomies, 4 segmental resections and one bilobectomy. Histological findings were: squamous cell carcinoma in 38 cases, adenocarcinoma in 15 and large cell carcinoma in three. In all 56 cases, invasive mucosal carcinoma was found exclusively on the bronchial resection margin. Nineteen tumours were stage I; 12, stage II; 17, stage IIIa; 5, stage IIIb; and three, stage IV. Nineteen patients (59.3%) with early stage tumours (I and II) received adjuvant radiation therapy and only three chemotherapy. RESULTS: The prognosis in these cases was disease-stage related (21 and 38.4% of deaths due to the disease). Forty-one percent of the stage IIIa patients received radiation therapy and 17.6% chemotherapy: 70.6% died of tumour relapse. Forty percent of the stage IIIb patients received radiation therapy and 20% chemotherapy: 60% died of disease progression. All of the stage IV patients died within 3 months from surgical resection. At the end of the study, 21 patients were alive after an interval of 22-142 months (18 in stage I or II). The 10-year actuarial survival rate was 44%. The percentage survival for stage IIIa was 16.8, after 10 years, and fell to 45 months for stage IIIb. CONCLUSIONS: The prognosis of our stage I or II patients with microresidual tumour on the bronchial resection margin (R1) was similar to that of the patients in the same disease stage, whose resection was microscopically radical (R0) and the same was true of the patients in stage III. In patients with residual tumour cells on the bronchial stump we did not observe worsened long-term survivals. PMID- 12559348 TI - Predictors of clinical outcome following extended thymectomy in myasthenia gravis. AB - OBJECTIVE: Thymectomy remains as the optimal treatment of choice in patients with myasthenia gravis (MG), however, the selection criteria for surgery remains controversial. METHODS: We examined the data charts of patients with MG underwent extended thymectomy. We investigated the possible correlations between the clinicopathologic features and clinical outcomes, and analyzed the data to clarify the effect of prognostic factors on clinical outcome. RESULTS: A total of 61 patients with a mean age of 35.8 +/- 12.2 years (range, 13-66 years) were analyzed. The overall improvement/remission and clinical worsening rates were 81.9 and 18.1%, respectively. Ossermann stage (P = 0.011) and presence of mediastinal ectopic thymic tissue (P = 0.007) showed a significant correlation with the clinical outcome. Multivariate analysis confirmed Ossermann stage (P = 0.0158), and presence of mediastinal ectopic thymic tissue (P = 0.0100) as independent predictors on clinical outcome. CONCLUSION: Ossermann stage and the presence of mediastinal ectopic thymic tissue are potential predictors on clinical outcome in patients with MG undergoing extended thymectomy. PMID- 12559349 TI - Remodeling after right pneumonectomy. PMID- 12559350 TI - Spontaneous pneumoperitoneum after pleuroperitoneal shunt. PMID- 12559351 TI - Bullous herniation of the lung through an intercostal drain site. PMID- 12559352 TI - Inferior vena cava leiomyosarcoma. PMID- 12559353 TI - Distant endoarterial bullet migration following penetrating chest injury. PMID- 12559354 TI - Pseudoaneurysm of the mammary to left anterior descending artery anastomosis after off-pump CABG. PMID- 12559355 TI - Staphylococcal postoperative subannular left ventricular false aneurysm. AB - A 37-year-old male with acute complicated methicillin-sensitive Staphylococcus aureus mitral valve endocarditis underwent urgent valve replacement with a bileaflet prosthesis. The postoperative course was complicated with fever and heart failure. Echocardiography showed a large subannular false aneurysm of the left ventricle. Three weeks later resection and closure of the defect with a patch made from a cryopreserved thoracic aorta homograft were performed. The most significant aspects of this rare complication are commented on. PMID- 12559356 TI - Closure of recurrent VSD due to dehiscence of calcified patch. AB - Closure of residual ventricular septal defects may be extremely difficult in the presence of severe calcification of a previous patch. Removal of such calcification carries a risk of damaging the aortic and tricuspid valve as well as the conduction system. We describe a novel technique for closure of such a defect placing a new patch over and around the calcified one in an 18-year-old patient who had undergone initial surgery 12 years ago. PMID- 12559357 TI - Superior mediastinal chordoma presenting as a bilobed paravertebral mass. AB - Thoracic chordomas are rare, low-grade malignant tumours arising from the notochordal remnants. These tumours are locally invasive and hence have a tendency for frequent local recurrence. This article presents an unusual appearance of a chordoma, as a bilobed tumour involving both the hemithoraxes. Since the treatment of choice for these tumours is complete surgical excision this involved multiple surgical procedures. PMID- 12559358 TI - Synchronous bilateral typical pulmonary carcinoid tumors. AB - A 73-year-old woman was admitted with bilateral pulmonary nodules, one on each side, which were suspected to be metastases of an unknown primary tumor. Enucleation of the lesion in the upper right lobe was performed. The histological examination showed a typical carcinoid tumor. A subsequent octreotid scan revealed high suspicion of a carcinoid in the left lower lobe as well. Formal right upper lobectomy and left lower lobectomy were performed in staged procedures. The radiological 1 year control with CT-scan demonstrated no further pathological lesions. To the best of our knowledge this is the first description of curable synchronous pulmonary carcinoid tumors. PMID- 12559360 TI - Resection of locally advanced thymic carcinoid tumors. PMID- 12559361 TI - The management of malignant pleural mesothelioma. PMID- 12559363 TI - Intrathoracic staging of non small cell lung cancer with mediastinal involvement: still a moonless foggy night? PMID- 12559365 TI - Primary hydatid cysts of the mediastinum. PMID- 12559367 TI - MDR1-deficient genotype in Collie dogs hypersensitive to the P-glycoprotein substrate ivermectin. AB - Multidrug resistance (MDR) phenotypes in cancer cells are associated with overexpression of the drug carrier P-glycoprotein. The antiparasitic drug ivermectin, one of its substrates, abnormally accumulates in the brain of transgenic mice lacking the P-glycoprotein, resulting in neurotoxicity. Similarly, an enhanced sensitivity to ivermectin has been reported in certain dogs of the Collie breed. To explore the basis of this phenotype, we analyzed the canine P-glycoprotein-encoding MDR1 gene, and we report the first characterization of the cDNA for wild-type (Beagle) P-glycoprotein. The corresponding transcripts from ivermectin-sensitive Collies revealed a homozygous 4-bp exonic deletion. We established, by genetic testings, that the MDR1 frame shift is predictable. Accordingly, no P-glycoprotein was detected in the homozygote-deficient dogs. In conclusion, we characterized a unique case of naturally occurring gene invalidation. This provides a putative novel model that remains to be exploited in the field of human therapeutics and that might significantly affect tissue distribution and drug bioavailability studies. PMID- 12559368 TI - DIF-1, an anti-tumor substance found in Dictyostelium discoideum, inhibits progesterone-induced oocyte maturation in Xenopus laevis. AB - Differentiation-inducing factor-1 (DIF-1; 1-(3,5-dichloro-2,6-dihydroxy-4 methoxyphenyl)hexan-1-one) is a putative morphogen that induces stalk-cell formation in the cellular slime mold Dictyostelium discoideum. DIF-1 has previously been shown to suppress cell growth in mammalian cells. In this study, we examined the effects of DIF-1 on the progesterone-induced germinal vesicle breakdown in Xenopus laevis, which is thought to be mediated by a decrease in intracellular cAMP and the subsequent activation of mitogen-activated protein kinase (MAPK) and maturation-promoting factor, a complex of cdc2 and cyclin B, which regulates germinal vesicle breakdown. DIF-1 at 10-40 microM inhibited progesterone-induced germinal vesicle breakdown in de-folliculated oocytes in a dose-dependent manner. Progesterone-induced cdc2 activation, MAPK activation, and c-Mos accumulation were inhibited by DIF-1. Furthermore, DIF-1 was found to inhibit the progesterone-induced cAMP decrease in the oocytes. These results indicate that DIF-1 inhibits progesterone-induced germinal vesicle breakdown possibly by blocking the progesterone-induced decrease in [cAMP](i) and the subsequent events in Xenopus oocytes. PMID- 12559369 TI - Enhancement of antinociception by co-administration of an opioid drug (morphine) and a preferential cyclooxygenase-2 inhibitor (rofecoxib) in rats. AB - Synergism has been used to obtain analgesia at doses at which side effects are minimal. In addition, it has been demonstrated that inhibition of cyclooxygenase 2 is responsible for the therapeutic effects of nonsteroidal anti-inflammatory drugs (NSAIDs). The aim of this study was to evaluate the antinociceptive interaction between the preferential COX-2 inhibitor, rofecoxib and morphine. Several combinations were evaluated using the pain-induced functional impairment model (PIFIR), a rat model of arthritic pain. Surface of synergistic interaction (SSI) analysis and an isobolographic method were used to detect the antinociceptive potency of the drugs, given either individually or in combination. The surface of synergistic interaction was calculated from the total antinociceptive effect produced by the combination after subtraction of the antinociceptive effect produced by each individual drug. Male rats received orally morphine alone (10, 17.8, 31.6, 56.2 and 100.0 mg/kg), rofecoxib alone (3.2, 5.6, 10, 31.6, 56.2 and 74.0 mg/kg) or 12 different combinations of morphine and rofecoxib. Three combinations exhibited potentiation of antinociceptive effects (10 mg/kg of morphine with either 5.6, 10 or 31.6 mg/kg of rofecoxib), whereas the other nine combinations showed additive antinociceptive effects. The combination of morphine, 56.2 mg/kg (p.o.), and rofecoxib, 31.6 mg/kg (p.o.), produced the maximum antinociceptive effect (P<0.05). This combination caused gastric injuries less severe than those observed with indomethacin, i.e. it reduced ulcers and erosion formation. The synergistic antinociceptive effects of rofecoxib and morphine are important and suggest that combinations with drugs may decrease the side effects associated with the use of nonselective NSAIDs. Furthermore, the present results suggest that combinations containing opioid drugs and selective COX-2 inhibitors may have clinical utility in pain therapy. PMID- 12559370 TI - The anticonvulsant retigabine attenuates nociceptive behaviours in rat models of persistent and neuropathic pain. AB - We have tested for anti-nociceptive effects of the anticonvulsant KCNQ channel opener, N-(2-amino-4-(4-fluorobenzylamino)-phenyl)carbamic acid ethyl ester (retigabine), in rat models of experimental pain. In the chronic constriction injury and spared nerve models of neuropathic pain, injection of retigabine (5 and 20 mg/kg, p.o.) significantly attenuated (P<0.05) mechanical hypersensitivity in response to pin prick stimulation of the injured hindpaw. In contrast, retigabine had no effect on mechanical hypersensitivity to von Frey stimulation of the injured hindpaw in either model. Cold sensitivity in response to ethyl chloride was only attenuated (P<0.05) in the chronic constriction injury model. In the formalin test, retigabine (20 mg/kg, p.o.) attenuated flinching behaviour in the second phase compared with vehicle (P<0.05), and this effect was completely reversed by the KCNQ channel blocker 10,10-bis(4-pyridinylmethyl) 9(10H)-anthracenone (XE-991; 3 mg/kg, i.p.). Neither retigabine nor XE-991 administration affected the latency to respond to noxious thermal stimulation of the tail in control animals. These results suggest that retigabine may prove to be effective in the treatment of neuropathic pain. PMID- 12559371 TI - Role of muscarinic receptors in the activation of the ventral subiculum and the consequences for dopamine release in the nucleus accumbens. AB - The nucleus accumbens receives limbic inputs from a number of brain regions, including the ventral subiculum. In rats, activation of the ventral subiculum following microinjection of N-methyl-D-aspartate (NMDA) or carbachol increases locomotor activity, whilst ventral subiculum application of NMDA also increases dopamine efflux in the ipsilateral nucleus accumbens. Microdialysis experiments were therefore conducted to ascertain the consequences for dopamine release in the nucleus accumbens following ventral subiculum administration of carbachol, and to explore the acetylcholine receptor subtype(s) that might be involved. We report that, in anaesthetised rats, ventral subiculum administration of carbachol increased dopamine levels in the nucleus accumbens. The response was attenuated by co-administration with atropine, whilst administration of nicotine and the alpha-7 nicotinic acetylcholine receptor agonist AR-R17779 (spiro[1 azabicyclo[2,2,2]octane-3,5'-oxazolidine]-2'-one monohydrochloride) failed to evoke a response. Oxotremorine-M produced a dose-dependent increase in dopamine efflux confirming sensitivity to muscarinic receptor stimulation. However, the ventral subiculum was insensitive to xanomeline and pilocarpine, muscarinic M(1) receptor-preferring agonists, but sensitive to BuTAC ([5R-[exo]-6-[butylthio] 1,2,5-thiadiazol-3-yl]-1-azabicyclo[3.2.1])octane), a muscarinic M(2)/M(4) receptor agonist. The dopamine response to oxotremorine-M was significantly attenuated, although not abolished by co-administration with the M(2)/M(4) receptor antagonist methoctramine, and studies combining oxotremorine-M with (-) bicuculline, indicated a dual action in the ventral subiculum that was dependent and independent of reduced GABA neurotransmission. The data presented indicates that activation of the ventral subiculum by carbachol increases dopamine efflux in the nucleus accumbens by stimulation of muscarinic receptors, and that the ventral subiculum-nucleus accumbens projection system is sensitive to muscarinic M(2)/M(4) receptor stimulation. PMID- 12559372 TI - Influence of prazosin and clonidine on morphine analgesia, tolerance and withdrawal in mice. AB - Rapid development of tolerance and dependence limits the usefulness of morphine in long-term treatment. We examined the effects of clonidine (alpha(2) adrenoceptor agonist) and prazosin (alpha(1)-adrenoceptor antagonist) on morphine analgesia, tolerance and withdrawal. Morphine tolerance was induced using a 3-day cumulative twice-daily dosing regimen with s.c. doses up to 120 mg/kg. Tolerance was assessed on day 4, as loss of the antinociceptive effect of a test dose of morphine (5 mg/kg). After 10 h, morphine withdrawal was precipitated with naloxone (1 mg/kg). Prazosin had no analgesic effect alone but dose-dependently potentiated morphine analgesia in morphine-naive mice. Another alpha(1) adrenoceptor antagonist, corynanthine, had similar effects. Prazosin also increased the analgesic potency of the morphine test dose in morphine-tolerant mice. Naloxone-precipitated vertical jumping was not affected, but weight loss was reduced by prazosin. Acutely administered clonidine potentiated morphine analgesia and alleviated opioid withdrawal signs, as expected. We conclude that in addition to the already established involvement of alpha(2)-adrenoceptors in opioid actions, also alpha(1)-adrenoceptors have significant modulatory role in opioid analgesia and withdrawal. PMID- 12559373 TI - Central injection of astressin inhibits carbon tetrachloride-induced acute liver injury in rats. AB - The effect of intracisternal astressin, a specific and potent corticotropin releasing factor (CRF)(1) and CRF(2) receptor antagonist on carbon tetrachloride (CCl(4))-induced acute liver injury was investigated in rats. Intracisternal astressin inhibited the elevation of serum alanine aminotransferase level induced by CCl(4). Intracisternal astressin also reduced CCl(4)-induced liver histological changes. The protective effect of central astressin on CCl(4) induced liver damage was abolished by sympathectomy but not by hepatic branch vagotomy. These findings demonstrate that astressin acts in the central nervous system to induce hepatic cytoprotection, possibly through the sympathetic pathways in rats. These results further establish a role of endogenous CRF in the brain in hepatic pathophysiological regulation. PMID- 12559374 TI - Effects of short-acting hypnotics on sleep latency in rats placed on grid suspended over water. AB - The present study was performed to develop a new sleep disturbance model for evaluating hypnotic potencies by placing rats on a grid suspended over water up to 1 cm under the grid surface. When rats were placed on the grid, significant increases in sleep latency and amount of wakefulness were observed compared with those of rats placed on sawdust. However, the amounts of non-rapid eye movement (non-REM) sleep and rapid eye movement (REM) sleep of rats placed on the grid were significantly decreased compared with those of rats placed on sawdust. Four short-acting hypnotics (triazolam, zopiclone, brotizolam, lormetazepam) caused significant decreases in sleep latency, and the effects of hypnotics in rats placed on the grid were more potent than those in rats placed on sawdust. In conclusion, the present model can serve as a new sleep disturbance model and may also be useful for evaluating the sleep-inducing effects of short-acting hypnotics. PMID- 12559375 TI - Vasoconstrictor responsiveness of tail arteries from endotoxaemic rats. AB - Continuous infusion of lipopolysaccharide in conscious rats mimics some aspects of cardiovascular dysfunction in septic shock. In the present study, contractile responsiveness of tail arteries taken from rats infused with lipopolysaccharide was investigated. Contractile responses to alpha,beta-methylene ATP and potassium chloride, but not to methoxamine, were greater after 24 h lipopolysaccharide infusion than in 2-h saline, 24-h saline and 2-h lipopolysaccharide groups. N(G) nitro-L-arginine methyl ester augmented contractions to alpha,beta-methylene ATP and methoxamine in the 2-h saline, 24-h saline and 2-h lipopolysaccharide groups, but had no significant effect in the 24-h lipopolysaccharide group. Endothelium independent vasorelaxant responses to sodium nitroprusside were greater in the 24 h lipopolysaccharide group compared to the other three groups. Relaxations to acetylcholine were not significantly different. In vitro incubation in medium containing lipopolysaccharide for 24 h had no significant effect on contractile responses of tail arteries compared to controls incubated in medium alone. These data indicate a possible impaired nitric oxide and/or endothelial function in tail arteries isolated from rats 24 h after lipopolysaccharide infusion. As hypercontractility was not evoked following in vitro incubation with lipopolysaccharide, the involvement of in vivo neurohumoral factors/mechanisms in the pathology of these changes is implicated. PMID- 12559376 TI - Dobutamine administration exacerbates postischaemic myocardial dysfunction in isolated rat hearts: an effect reversed by thyroxine pretreatment. AB - The present study has investigated the effects of dobutamine on postischaemic dysfunction in the setting of global ischaemia and reperfusion in a model of isolated heart preparation. Isolated rat hearts were subjected to 20 min of zero flow global ischaemia followed by 45 min of reperfusion. Dobutamine administration (10 microg/kg/min) during the reperfusion period resulted in deterioration of functional recovery, which was abolished by propranolol administration. Long-term thyroxine pretreatment (12.5 microg 100 g(-1) body weight, b.i.d., s.c., for 2 weeks) reversed the detrimental effect of dobutamine and increased postischaemic recovery of function. We conclude that the combination of thyroxine pretreatment and dobutamine administration could potentially be a new therapeutic strategy to improve postischaemic dysfunction particularly in clinical settings such as cardiopulmonary bypass and/or myocardial infarction. PMID- 12559377 TI - Kinetic inhibitory profile of BIA 3-202, a novel fast tight-binding, reversible and competitive catechol-O-methyltransferase inhibitor. AB - The present study reports the kinetic inhibitory profile of 1-[3,4-dihydroxy-5 nitrophenyl]-2-phenyl-ethanone (BIA 3-202), a novel inhibitor of soluble catechol O-methyltransferase (COMT) in rat liver. After an oral single dose (30 mg kg( 1)), there was a time-dependent recovery of enzyme activity from 98+/-2% inhibition at 30 min to total recovery at 24 h after treatment. The inhibitory effect produced by BIA 3-202 on soluble COMT was reversible after gel filtration of the samples. BIA 3-202 acted as a fast inhibitor of rat liver soluble COMT, interacting immediately with the enzyme after mixing. No differences were observed in the metanephrine formation rates (in nmol mg protein(-1) min(-1)) obtained without and with a 60-min preincubation with 30 nM of BIA 3-202 (1.30+/ 0.02 and 1.35+/-0.03, respectively). The tight-binding nature of the inhibition produced by BIA 3-202 was evaluated by performing an Ackermann-Potter plot. The true K(i) for BIA 3-202, derived from the nonlinear regression analysis, was 0.19+/-0.02 nM. In substrate competition studies, an increase in the concentration of adrenaline resulted in a linear increase in IC(50) values for BIA 3-202. In conclusion, BIA 3-202 behaves as a reversible, potent and fast tight-binding COMT inhibitor that acts competitively at the substrate binding site of rat liver soluble COMT. PMID- 12559378 TI - Dosing time-dependent variation in the hypocalcemic effect of calcitonin in rat. AB - Dosing time-dependent variation in the hypocalcemic effect of salmon calcitonin was examined in rats under a 12-h light-dark cycle. In both a single-dosing study with normal rats and a repeated-dosing study with hypercalcemic rats (induced by chronic vitamin-D dosing), we consistently observed that the hypocalcemic effect of calcitonin was greater when the drug was given at 14 h after lights on than that at 2 h after lights on. The reduction in urinary deoxypyridinoline excretion, a marker of bone resorption, was also greater when calcitonin was given at 14 h after lights on. Urinary excretion of Ca was not affected by the drug. Pharmacokinetic profiles of calcitonin after a single dosing did not differ between the two trials. These results indicate that the hypocalcemic effect of calcitonin is greater after dosing in the early dark phase (14 h after lights on) than after dosing in the early light phase (2 h after lights on). A time dependent variation in the sensitivity to the drug of osteoclasts, but not renal tissues, may be involved in the mechanism of this event. PMID- 12559379 TI - Morphine as a drug for stress ulcer prevention and healing in the stomach. AB - Morphine pretreatment protects against stress-induced gastric ulceration, however, the exact mechanism is still undefined. Interestingly, the effect of morphine on ulcer healing has not been investigated. In this report, we would like to study these effects in a defined stress ulcer model and to delineate a new implication for morphine to promote stress ulcer healing in rats. Our study showed that cold-restraint stress for 3 h induced hemorrhagic lesions and increased myeloperoxidase activity in the gastric mucosa. Stress also reduced the dimension of layer of periodic acid-Schiff reagent-stained cells in the gastric mucosa by about 50%. Morphine pretreatment (2 or 8 mg/kg, given intraperitoneally) at the time of stress dose-dependently reversed stress-induced gastric ulceration, increase of myeloperoxidase activity and reduction of thickness of mucus-stained cells in the gastric mucosa. Morphine treatment after stress (given at the end of a 3-h stress and also at 3 h thereafter) increased ulcer healing by reducing the ulcer size measured 24 h later. Such action was blocked by naloxone (8 mg/kg) given intraperitoneally 15 min before morphine treatment. Morphine also increased the number of cell proliferation and dimension of layer of cells stained for mucus but not the number of microvessels in the gastric mucosa. Moreover, the number of apoptotic cells was less evidenced in the morphine-treated rats. This study reports for the first time that morphine not only prevents stress ulceration but also promotes healing of stress ulcer through a defined mechanism. PMID- 12559380 TI - Neurotransmitters involved in the fast inhibitory junction potentials in mouse distal colon. AB - We investigated, in murine colon circular muscle, the role of adenosine 5' triphosphate (ATP) and pituitary adenylate cyclase activating peptide (PACAP) as inhibitory neurotransmitters of the fast component of nerve-evoked inhibitory junction potential (fast IJP). Fast IJP was antagonised by apamin or suramin, abolished by desensitisation with the P2Y receptor agonist, adenosine 5'-O-2 thiodiphosphate (ADPbetaS), unaffected by desensitisation with P2X receptor agonist, alpha,beta-methylene ATP (alpha,beta-meATP), and reduced by PACAP-(6 38), a PACAP receptor antagonist. ATP induced membrane hyperpolarization resistant to tetrodotoxin, N(omega)-nitro-L-arginine methyl ester (L-NAME) or PACAP-(6-38), but antagonised by apamin, suramin, P2X and P2Y receptor desensitisation. PACAP-(1-27) caused membrane hyperpolarization antagonised by PACAP-(6-38), apamin and P2Y receptor desensitisation, reduced by tetrodotoxin, but not affected by L-NAME and by P2X receptor desensitisation. Therefore, in murine colon circular muscle, an ATP-like endogenous P2Y purinoceptor ligand is the final nonadrenergic, noncholinergic (NANC) inhibitory mediator involved in the generation of fast IJP. A neuromodulator role of PACAP in the inhibitory pathway is supposed. PMID- 12559381 TI - Regional differences in the adenosine A(2) receptor-mediated modulation of contractions in rat vas deferens. AB - Adenosine receptors involved in modulation of contractions were characterized in the bisected rat vas deferens by combining pharmacological and immunohistochemical approaches. In both portions, noradrenaline-elicited contractions were enhanced by the adenosine A(1) receptor agonist N(6) cyclopentyladenosine (CPA), and inhibited by the non-selective adenosine receptor agonist 5'-N-ethylcarboxamidoadenosine (NECA) in the presence of the adenosine A(1) receptor antagonist 1,3-dipropyl-8-cyclopentyl-l,3-dipropylxanthine (DPCPX). The adenosine A(2A) receptor agonist 2-p-(2-carboxyethyl)phenethyl-amino-5'-N ethylcarboxamidoadenosine (CGS 21680) also inhibited noradrenaline-elicited contractions but only in the prostatic portion. Contractions elicited by the stable ATP analogue alpha,beta-methyleneATP (alpha,beta-MeATP) were inhibited only by NECA in the presence of DPCPX and only in the prostatic portion. This study provides functional evidence for the presence, in both portions of the rat vas deferens, of an adenosine A(1) receptor-mediated enhancement and of an adenosine A(2) receptor-mediated inhibition of contractions. The latter effect is mediated by both A(2A) and A(2B) subtypes in the prostatic portion but only by the A(2B) subtype in the epididymal portion. This regional variation is supported by the immunohistochemical results that revealed an adenosine A(2A) receptor immunoreactivity not co-localized with nerve fibres more abundant in the prostatic than in the epididymal portion. PMID- 12559382 TI - Role of opioid and nitric oxide systems in the nonadrenergic noncholinergic mediated relaxation of corpus cavernosum in bile duct-ligated rats. AB - Changes in nonadrenergic noncholinergic (NANC)-mediated relaxation of the anococcygeus muscle have been demonstrated in cholestasis. Cholestasis is also associated with accumulation of endogenous opioid peptides and nitric oxide (NO) overproduction. This study was therefore undertaken to investigate the effect of cholestasis on the NANC-mediated relaxation of corpus cavernosum in bile duct ligated rats and to examine the possible roles of the opioid system and nitric oxide in the cholestasis-associated alterations of corpus relaxation. Bile duct ligated and sham-operated rats were treated for 2 weeks with either normal saline, N (omega)-nitro L-arginine methylester (L-NAME) (3 mg/kg/day, i.p.) or naltrexone (20 mg/kg/day, i.p.). On the 14th day, the strips of corpus cavernosum were mounted under tension in a standard oxygenated organ bath with guanethidine sulfate (5 microM) and atropine sulfate (1 microM) (to produce adrenergic and cholinergic blockade). The strips were precontracted with phenylephrine hydrochloride (7.5 microM) and electrical field stimulation was applied at different frequencies to obtain NANC-mediated frequency-dependent relaxant responses. The results showed that the amplitudes of relaxation responses at each frequency in bile duct-ligated rats were greater than the responses of sham operated animals. This increase in relaxation responses in bile duct-ligated rats was inhibited by chronic L-NAME administration for 2 weeks so it seemed that it might be due to the nitric oxide overproduction in cholestatic states. Chronic administration of naltrexone for 2 weeks to bile duct-ligated rats had the same inhibitory effect on the relaxation responses. Our results demonstrated that in cholestasis, there was an increase in NANC-mediated relaxation of corpus cavernosum and both opioid and nitric oxide systems were involved in this increase. PMID- 12559383 TI - Effects of dosmalfate, a new cytoprotective agent, on acute and chronic trinitrobenzene sulphonic acid-induced colitis in rats. AB - Activated neutrophils and proinflammatory cytokines, such as tumor necrosis factor-alpha (TNF-alpha) are clearly involved in the pathogenesis of bowel disease. Increased expression of epidermal growth factor-receptor (EGF receptor) has been reported for the colon mucosa surrounding areas of ulceration, suggesting a pivotal role in mucosal defence and repair. In this study, we examined the effects of dosmalfate, a new flavonoid derivative compound (diosmin heptakis) with antioxidant and cytoprotective properties, on acute and chronic experimental trinitrobenzene sulphonic acid (TNBS)-induced colitis in rats. The inflammation response was assessed by neutrophil infiltration as evaluated by histology and myeloperoxidase activity. Mucosal TNF-alpha production and histological analysis of the lesions was also carried out. In addition, we studied the expression of the EGF receptor inmunohistochemically during the healing of TNBS-induced chronic colitis. A 2-day treatment with 400 or 800 mg/kg of dosmalfate ameliorated the colon damage score and the incidence of adhesions. It also significantly (P<0.05) decreased myeloperoxidase activity and colonic mucosal production of TNF-alpha. Chronic treatment (14 days) with 800 mg/kg/day of dosmalfate also had significant protective effects on TNBS-induced colitis which were reflected by significant attenuation (P<0.05) of the damage score while the inflammatory indicators were not improved. The chronic beneficial effect of dosmalfate was apparently related to the enhancement of EGF receptor expression. These findings confirm the protective effects of dosmalfate in acute and chronic experimental colitis. PMID- 12559384 TI - A new dual inhibitor of arachidonate metabolism isolated from Helichrysum italicum. AB - Six acetophenones (1-6) and one gamma-pyrone (7), previously isolated from Helichrysum italicum, were tested for their ability to inhibit enzymatic and non enzymatic lipid peroxidation, the stable 1,1-diphenyl-2-pycryl-hydrazyl free radical, superoxide scavenging and arachidonic acid metabolism. In addition, they were studied in different experimental models such as the chronic inflammation induced by 12-O-tetradecanoylphorbol 13-acetate (TPA), the phospholipase A(2) induced mouse paw oedema test, the carrageenan-induced mouse paw oedema test, and the writhing induced by acetic acid in the mouse. Of the assayed compounds, only 1 inhibited enzymatic lipid peroxidation but had no effect on non-enzymatic lipid peroxidation. None of them scavenged the superoxide radical. Study of the inhibition of arachidonic acid metabolism demonstrated that 1 was an inhibitor of both cyclooxygenase and 5-lipoxygenase, whereas 2 was a selective inhibitor of 5 lipoxygenase. In the assay of phospholipase A(2)-induced mouse paw oedema, the gamma-pyrone derivative inhibited oedema formation, showing a similar profile to that obtained with cyproheptadine. The acetophenones were effective at 30 and 60 min. In the carrageenan test, acetophenone 1 gave the best results and had analgesic effects in the acetic acid writhing test. In conclusion acetophenone 1 (4-hydroxy-3-(3-methyl-2-butenyl)acetophenone) is a new dual inhibitor of arachidonate metabolism, and could be a useful tool for obtaining anti inflammatory and analgesic drugs. PMID- 12559385 TI - Regulators of G-protein signalling: multifunctional proteins with impact on signalling in the cardiovascular system. AB - Regulator of G-protein signalling (RGS) proteins form a superfamily of at least 25 proteins, which are highly diverse in structure, expression patterns, and function. They share a 120 amino acid homology domain (RGS domain), which exhibits GTPase accelerating activity for alpha-subunits of heterotrimeric G proteins, and thus, are negative regulators of G-protein-mediated signalling. Based on the organisation of the Rgs genes, structural similarities, and differences in functions, they can be divided into at least six subfamilies of RGS proteins and three more families of RGS-like proteins. Many of these proteins regulate signalling processes within cells, not only via interaction with G protein alpha-subunits, but are G-protein-regulated effectors, Gbetagamma scavenger, or scaffolding proteins in signal transduction complexes as well. The expression of at least 16 different RGS proteins in the mammalian or human myocardium have been described. A subgroup of at least eight was detected in a single atrial myocyte. The exact functions of these proteins remain mostly elusive, but RGS proteins such as RGS4 are involved in the regulation of G(i) protein betagamma-subunit-gated K(+) channels. An up-regulation of RGS4 expression has been consistently found in human heart failure and some animal models. Evidence is increasing that the enhanced RGS4 expression counter regulates the G(q/11)-induced signalling caused by hypertrophic stimuli. In the vascular system, RGS5 seems to be an important signalling regulator. It is expressed in vascular endothelial cells, but not in cultured smooth muscle cells. Its down-regulation, both in a model of capillary morphogenesis and in an animal model of stroke, render it a candidate gene, which may be involved in the regulation of capillary growth, angiogenesis, and in the pathophysiology of stroke. PMID- 12559386 TI - L-3,4-Dihydroxyphenylalanine as a neurotransmitter candidate in the central nervous system. AB - Historically, 3,4-dihydroxyphenylalanine (DOPA) has been believed to be an inert amino acid that alleviates the symptoms of Parkinson's disease by its conversion to dopamine via the enzyme aromatic L-amino acid decarboxylase. In contrast to this generally accepted idea, we propose that DOPA itself is a neurotransmitter and/or neuromodulator, in addition to being a precursor of dopamine. Several criteria, such as synthesis, metabolism, active transport, existence, physiological release, competitive antagonism, and physiological or pharmacological responses, must be satisfied before a compound is accepted as a neurotransmitter. Recent evidence suggests that DOPA fulfills these criteria in its involvement mainly in baroreflex neurotransmission in the lower brainstem and in delayed neuronal death by transient ischemia in the striatum and the hippocampal CA1 region of rats. PMID- 12559387 TI - The many roles of cytochrome b5. AB - Four distinct suggestions have been made to explain the mechanism of the cytochrome b(5)-imposed positive modifier action of the cytochrome P450 monooxygenase reaction. The first mechanism involves a direct input of an electron into the monooxygenase cycle. This is the second of the two electrons necessary for activation of molecular oxygen, and appears to be a rate-limiting step in the monooxygenase reaction. P450 monooxygenases all appear to be uncoupled to varying extents, releasing superoxide and hydrogen peroxide instead of oxidized substrate. A second mechanism suggests that cytochrome b(5) acts as a positive modifier of the monooxygenase by decreasing the extent of uncoupling of the monooxygenase reaction. The implication is that a slow input of the second electron allows uncoupling of a superoxide anion instead of formation of two electron reduced oxygen. Faster input of the second electron via cytochrome b(5) would result in formation of more of the activated oxygen that reacts with substrate to form product. A third suggestion involves formation of a two hemoprotein complex between cytochrome b(5) and cytochrome P450 that allows acceptance of two electrons from NADPH-cytochrome P450 reductase. Uncomplexed cytochrome P450 accepts an electron from the reductase, dissociates from it, binds oxygen, and re-associates with the reductase to accept another electron. Complexation with cytochrome b(5) enhances the rate of formation of the active oxygen by obviating the need for two interactions with reductase. The fourth mechanism has cytochrome b(5) serving as an effector without a reduction oxidation role in the monooxygenation reaction. This effector function may be to enhance the breakdown of the oxygenated hemoprotein to products or to facilitate flow of electrons through the system. PMID- 12559389 TI - Regulation of 5-HT1A receptor function in brain following agonist or antidepressant administration. AB - Adaptive changes in the serotonergic system are generally believed to underlie the therapeutic effectiveness of the azapirone anxiolytics and a variety of antidepressant drugs. The serotonin-1A (5-HT(1A)) receptor has been implicated in affective disorders. Thus, studies of the regulation of 5-HT(1A) receptor function may have important implications for our understanding the role of this receptor in the mechanism of action of these therapeutic agents. This review focuses on the regulation of central 5-HT(1A) receptor function following administration of 5-HT(1A) receptor agonists or antidepressant drugs expected to increase the synaptic concentration of the neurotransmitter 5-HT. The majority of evidence supports regional differences in the regulation of central 5-HT(1A) receptor function following repeated agonist or antidepressant administration, which may be due to differences in processes involved in desensitization of the receptor at the cellular level. Region-specific differences in the regulation of 5-HT(1A) receptor function may be based on compensatory changes distal to the receptor, such as regulatory changes at the level of effector (e.g. adenylyl cyclase or ion channel), or at the level of the G protein such as changes in G protein expression, or phosphorylation of the G protein. It may be that the increase in serotonin neurotransmission, due to somatodendritic autoreceptor desensitization following agonist or antidepressant treatment, to normo-sensitive 5-HT(1A) receptors in certain brain regions (e.g. hippocampus or cortex) and to sub-sensitive 5-HT(1A) receptors in other brain regions (e.g. amygdala or hypothalamus) underlies the therapeutic efficacy of these drugs. PMID- 12559390 TI - Over expression of bone morphogenetic protein-3b (BMP-3b) using an adenoviral vector promote the osteoblastic differentiation in C2C12 cells and augment the bone formation induced by bone morphogenetic protein-2 (BMP-2) in rats. AB - BMP-3b is a novel BMP-3-related protein and its biological functions are unknown. In order to investigate the biological actions of BMP-3b, we constructed a BMP-3b expressing recombinant adenoviral vector (AxCAKBMP-3b). We show that over expression of BMP-3b stimulated the induction of differentiation and the osteoinduction activity of a human BMP-2-expressing recombinant adenoviral vector (AxCAOBMP-2). C2C12 cells were infected in vitro with AxCAKBMP-3b, AxCAOBMP-2 and a control vector containing no foreign genes (AxCAwt). Cells infected with AxCAOBMP-2 and AxCAKBMP-3b produced more alkaline phosphatase and secreted more osteocalcin into the culture medium than cells infected with AxCAOBMP-2 and AxCAwt. When AxCAOBMP-2, AxCAKBMP-3b, and AxCAwt were injected into the calf muscles of nude rats (F 344/N Jcl-rnu), the osteoinduction seen with AxCAOBMP-2 and AxCAKBMP-3b was greater than that seen with AxCAOBMP-2 and AxCAwt. PMID- 12559391 TI - Cellular density regulation of plasminogen gene expression in mouse hepatocytes. AB - The liver produces a variety of proteins including plasminogen. Plasminogen is pro-enzyme that is converted into plasmin by plasminogen activator. Plasmin has a broad substrate spectrum and participates in several biological processes, such as fibrinolysis, tissue remodeling, cell migration, angiogenesis and embryogenesis. In the present study, the regulation of plasminogen expression in mouse hepatocytes was investigated in the primary culture system. Expression level of plasminogen mRNA in the culture at the low cell density condition (0.2 x 10(5) cells / cm(2)) was compared with that at the high cell density condition (1.0 x 10 (5) cells / cm(2)). In the low cell density culture, the expression level of plasminogen mRNA decreased by a time-dependent manner. However, mRNAs for albumin and alpha(2)-antiplasmin were not influenced by the low cell density culture. On the other hand, in the high cell density culture, plasminongen mRNA expressed constantly as well as albumin and alpha(2)-antiplasmin mRNAs. Thus, the decrease in plasminogen mRNA expression could specifically occur when the density of hepatocytes was low. The down-regulation of plasminogen mRNA in the low cell density culture is not observed in the presence of cycloheximide, suggesting that the de novo protein synthesis is required for the regulatory mechanism. These findings indicate that the expression of plasminogen mRNA from hepatocyte is dependent on the cell density and the stimulation by cell-cell contact may be an important factor for the constitutive expression of plasminogen gene in hepatocytes. PMID- 12559392 TI - Phyllanthus urinaria triggers the apoptosis and Bcl-2 down-regulation in Lewis lung carcinoma cells. AB - Phyllanthus urinaria (P. urinaria), a widely used herb medicine, was tested for the anticancer effect in its water extract for the first time. The water extract of P. urinaria significantly decreased the number of Lewis lung carcinoma cells in a dose-and time-dependent manner as determined by MTT assay. However, the water extract of P. urinaria did not exert any cytotoxic effect on normal cells such as endothelial cells and liver cells. Result from flow cytometry revealed a dose-dependent increase of dead cells 24 hours after treating Lewis lung carcinoma cells with P. urinaria extract. The anticancer activity of P. urinaria extract was due to the apoptosis induced in Lewis lung carcinoma cells, which was demonstrated by DNA fragmentation analysis and increased caspase-3 activity. The apoptosis triggered by P. urinaria extract in Lewis lung carcinoma cells was associated with the down-regulation of Bcl-2 gene expression, but not with p53, p21 and Bax. Furthermore, the partial inhibition of P. urinaria-induced apoptosis in Lewis lung carcinoma cells by pretreatment with cyclosporin A, a mitochondria permeability transition pore inhibitor, suggesting that P. urinaria extract induced the apoptosis of Lewis lung carcinoma cells, at least in part, through a mitochondria-associated intrinsic pathway. PMID- 12559393 TI - Changes in catecholamine metabolism by ascorbic acid deficiency in spontaneously hypertensive rats unable to synthesize ascorbic acid. AB - We have previously reported the establishment of a novel rat strain, SHR-od, with both spontaneous hypertension and a defect of ascorbic acid biosynthesis. Blood pressure in mature SHR-od fed an ascorbic acid-supplemented diet is over 190-200 mmHg, while it decreased to around 120 mmHg at 4-5 weeks after the cessation of ascorbic acid supplementation. With regard to possible mechanisms of blood pressure lowering, we focused on catecholamine synthesis in adrenal glands, since catecholamine is a major factor for blood pressure regulation and ascorbic acid is a co-factor of dopamine beta-hydroxylase (DBH) in catecholamine biosynthesis. Male SHR-od (25-week-old) and normotensive ODS rats with a defect in ascorbic acid biosynthesis (25-week-old) were fed a Funabashi-SP diet with or without ascorbic acid (300 mg/kg diet) for 28 days or 35 days. In SHR-od, systolic blood pressure (191 +/- 6 mmHg) began to decrease from day 21 in the ascorbic acid deficient group, whereas no significant difference was found in ODS rats. In spite of significant lowering of blood pressure, no significant differences were found in catecholamine levels in serum, adrenal glands and brain on day 28. On day 35, however, urinary excretion of norepinephrine and epinephrine in the ascorbic acid-deficient SHR-od were higher at 490% (P < 0.05) and 460% (P < 0.05) of the respective control. Serum catecholamine concentrations and the adrenal catecholamine content tended to be higher in the ascorbic acid-deficient SHR-od than the control of SHR-od and reached to similar level in ODS rats. The administration of ascorbic acid (intraperitoneal injection, 60 mg ascorbic acid/kg body weight, once a day) to the ascorbic acid-deficient SHR-od restored blood pressure to the range 180-190 mmHg within two days. These findings indicate that ascorbic acid deficiency affects catecholamine metabolism in the adrenal glands of SHR-od in response to blood pressure lowering, suggesting catecholamines are not involved in the mechanism for the remarkable reduction in blood pressure in response to ascorbic acid deficiency. PMID- 12559394 TI - Thiol oxidation by 2,2'-dithiodipyridine induced calcium mobilization in MG63 human osteosarcoma cells. AB - 2,2'-dithiodipyridine (2,2'-DTDP), a reactive disulphide that mobilizes Ca(2+) in muscle, induced an increase in cytoplasmic free Ca(2+)concentrations ([Ca(2+)](i)) in MG63 human osteosarcoma cells loaded with the Ca(2+)-sensitive dye fura-2. 2,2'-DTDP acted in a concentration-independent manner with an EC(50) of 50 microM. The Ca(2+) signal comprised an initial spike and a prolonged increase. Removing extracellular Ca(2+) did not alter the Ca(2+) signal, suggesting that the Ca(2+) signal was due to store Ca(2+) release. In Ca(2+)-free medium, the 2,2'-DTDP-induced [Ca(2+)](i) increase was not changed by depleting store Ca(2+) with 50 microM bredfeldin A (a Golgi apparatus permeabilizer), 2 microM carbonylcyanide m-chlorophenylhydrazone (CCCP, a mitochondrial uncoupler), 1 microM thapsigargin (an endoplasmic reticulum Ca(2+)pump inhibitor) or 5 microM ryanodine. Conversely, 2,2'-DTDP pretreatment abolished CCCP and thapsigargin induced [Ca(2+)](i) increases. 2,2'-DTDP-induced Ca(2+) signals in Ca(2+) containing medium were not affected by modulation of protein kinase C activity or suppression of phospholipase C activity. However, 2,2'-DTDP-induced Ca(2+) release was inhibited by a thiol-selective reducing reagent, dithiothreitol (5-25 microM) in a concentration-dependent manner. Collectively, this study shows that 2,2'-DTDP induced [Ca(2+)](i) increases in human osteosarcoma cells via releasing store Ca(2+)from multiple stores in a manner independent of protein kinase C or phospholipase C activity. The 2,2'-DTDP-induced store Ca(2+) release appeared to be dependent on oxidation of membranes. PMID- 12559395 TI - Evolutional study on acetylcholine expression. AB - Acetylcholine (ACh) is a well-known neurotransmitter in the cholinergic nervous systems of vertebrates and insects; however, there is only indirect evidence for its presence in lower invertebrates, such as plants and fungi. We therefore investigated the expression of ACh in invertebrates (sea squirt, sea urchin, trepang, squid, abalone, nereis, sea anemone, coral and sponge), plants (arabidopsis, eggplant, bamboo shoot, cedar, hinoki, pine, podcarp, fern, horsetail and moss), fungi (yeast and mushroom) and bacteria by assaying ACh content and synthesis, focusing on the presence of two synthetic enzymes, choline acetyltransferase (ChAT) and carnitine acetyltransferase (CarAT). Using a specific radioimmunoassay, ACh was detected in all samples tested. The levels varied considerably, however, with the upper portion of bamboo shoots having the highest content (2.9 micromol/g). ACh synthesis was also detected in all samples tested; moreover, the activity in most samples from the animal kingdom, as well as bamboo shoots and the stem of the shiitake mushroom, were sensitive to both ChAT and CarAT inhibitors. Levels of ACh synthesis were lower in samples from other plants, fungi and bacteria and were insensitive to ChAT and CarAT inhibitors. These findings demonstrate the presence of ACh and ACh-synthesizing activity in evolutionally primitive life as well as in more complex multicellular organisms. In the context of the recent discovery of non-neuronal ACh in various mammalian species, these findings suggest that ACh been expressed in organisms from the beginning of life, functioning as a local mediator as well as a neurotransmitter. PMID- 12559396 TI - Effects of acetazolamide and 4-aminoprydine on the responses of deflationary slowly adapting pulmonary stretch receptors to CO2 inhalation in the rat. AB - The inhibitory effect of CO(2) on deflationary slowly adapting pulmonary stretch receptors (deflationary SARs) was investigated before and after administration of acetazolamide, a carbonic anhydrase (CA) inhibitor, or 4-aminopyridine (4-AP), a K(+) channel blocker, in anesthetized, artificially ventilated rats after unilateral vagotomy. CO(2) inhalation (maximum tracheal CO(2) concentration ranging from 9 to 12%) for approximately 60 s decreased the impulse activity of deflationary SARs but had no significant effect on tracheal pressure (P(T)) as an index of bronchomotor tone. Acetazolamide treatment (20 mg/kg) diminished the inhibitory response of deflationary SARs to CO(2) inhalation. 4-AP (0.7 and 2.0 mg/kg) dose-dependently attenuated the decrease in deflationary SAR activity induced by CO(2) inhalation. When comparing the maximum attenuation due to 4-AP (2.0 mg/kg) and acetazolamide (20 mg/kg) in CO(2)-induced deflationary SAR inhibition, blockade of K(+) channels had a more pronounced effect. These results suggest that inhibition of deflationary SARs by CO(2) inhalation may be largely mediated by the stimulating action of 4-AP-sensitive K(+) currents in the nerve terminals of the receptors. PMID- 12559398 TI - Science of aging knowledge environment: one-stop shopping for researchers in the field of aging. AB - The Science of Aging Knowledge Environment (SAGE KE) was launched in October 2001 to provide an online information source and community-building tool. The site offers a wide range of features, including original commentary articles, a database of genes and interventions related to aging, and a calendar of meetings and events. Users may initiate discussions and post comments on the articles; these features are intended to promote interaction between researchers in the field and to ensure the timeliness of information posted. This paper details SAGE KE's contents and offers suggestions about how to customize the site to save time and maximize information acquisition and exchange. PMID- 12559397 TI - Melatonin prevents the increase in hydroxyl radical-spin trap adduct formation caused by the addition of cisplatin in vitro. AB - We studied the generation of reactive oxygen species (ROS) caused by cisplatin administration and the preventive effect of melatonin, the main secretory product of the pineal gland, on the reaction in vitro using electron spin resonance spectroscopy. Cisplatin induced generation of the hydroxyl radical (OH*) in phosphate buffer in pH 7.4 as a dose-dependent manner. However, OH* was not generated in phosphate buffer containing chloride ions at concentration exceeding 120 mM. The induction of OH. production by cisplatin was completely inhibited by the addition of melatonin, but not by the addition of 6-hydroxymelatonin, which is a hepatic metabolite of melatonin. Furthermore, melatonin was the most effective agent for preventing of OH* formation among various well-known antioxidants including mannitol and reduced glutathione. These results indicate that melatonin may scavenge OH. directly and thereby prevent renal tissue damage caused by OH* produced in response to cisplatin treatment. PMID- 12559399 TI - Experimental gerontological research in Australia. AB - Recently there has been recognition in Australia of the importance of experimental gerontological research. To date, the major areas of research into the biology of ageing have been oxidative stress and mitochondrial dysfunction. Future directions for gerontological research in Australia are likely to place more emphasis on comprehensive studies that integrate basic biological research, with clinical studies and the behavioural and social domains of ageing. Priority will be given to studies incorporating biological markers that are population based and longitudinal. The growing interest in experimental gerontological research complements Australian research strengths in other domains such as disease-based research, social gerontology, population studies and health policy research. PMID- 12559400 TI - Tokyo Metropolitan Institute of Gerontology. AB - The Tokyo Metropolitan Institute of Gerontology (TMIG) celebrates its 30th anniversary in 2002. Since its establishment, TMIG has been carrying out inter disciplinary research on aging as a core institute in Japan and functions as a knowledge bank and a source for capable researchers of gerontology. The proportion of people aged 65 and above in Japan will reach 30% in the near future, which might be associated with serious medical, social, political, and financial problems. Because most developed countries will face similar situations in the near future, multidisciplinary studies in gerontology on a worldwide scale are necessary to solve the associated problems. PMID- 12559401 TI - Aging of animals produced by somatic cell nuclear transfer. AB - Cloning by somatic cell nuclear transfer (NT) has been used to successfully generate viable mammalian embryos in many species. Despite the successful production of cloned animals, NT remains an inefficient technique at best. High rates of developmental failure and abnormalities are associated with NT-derived animals. Issues such as cell cycle regulation, epigenetic modification and oocyte activation have been examined in order to ultimately find ways to improve the efficiency of somatic cell NT. In reviewing the available literature regarding the generation of various mammalian species by using somatic cell NT, it becomes apparent that there are not only differences in the protocols used to generate such animals, but significant differences in the phenotypes of resultant animal clones as well. In this review we discuss the current findings about how aging and senescence are affected by somatic cell NT in mammals. PMID- 12559402 TI - Effect of aging on macrophage function. AB - In aged populations, infectious diseases are generally more serious and cause higher mortality. This has been attributed to a decline in the functional activity of the immune system. Many factors have been implicated, such as thymus involution and progressive impairment of the tissues and cells involved in the generation of immune response. One of the key constituents of the immune system are macrophages. In this review we present data produced over last 10 years about the effects of aging on macrophages. PMID- 12559388 TI - Molecular aspects of glutamate dysregulation: implications for schizophrenia and its treatment. AB - The glutamate system is involved in many aspects of neuronal synaptic strength and function during development and throughout life. Synapse formation in early brain development, synapse maintenance, and synaptic plasticity are all influenced by the glutamate system. The number of neurons and the number of their connections are determined by the activity of the glutamate system and its receptors. Malfunctions of the glutamate system affect neuroplasticity and can cause neuronal toxicity. In schizophrenia, many glutamate-regulated processes seem to be perturbed. Abnormal neuronal development, abnormal synaptic plasticity, and neurodegeneration have been proposed to be causal or contributing factors in schizophrenia. Interestingly, it seems that the glutamate system is dysregulated and that N-methyl-D-aspartate receptors operate at reduced activity. Here we discuss how the molecular aspects of glutamate malfunction can explain some of the neuropathology observed in schizophrenia, and how the available treatment intervenes through the glutamate system. PMID- 12559403 TI - Aging-related changes in the thiol/disulfide redox state: implications for the use of thiol antioxidants. AB - Genetic and biochemical studies suggest that free radical-derived reactive oxygen species play a key role in a common mechanism of aging in many or all animal species. This led to the hypothesis that the quality of life in old age may be improved by pharmacological or dietary thiol antioxidants. This review describes important details about how the organism deals with its own thiol antioxidants. Aging was found to be associated with an oxidative shift in the thiol/disulfide redox state (REDST) of the intracellular glutathione pool and of the plasma cyst(e)ine and albumin pools. There is also a decrease in plasma thiol (mainly cysteine) concentration. The oxidative shift in intracellular REDST was found to be typically associated with cellular dysfunctions. Studies in humans related to plasma REDST revealed correlations with aging-related pathophysiological processes, suggesting that oxidative changes in REDST play a key role in processes and diseases which limit the human life span. The age-related shift in plasma REDST is mediated, at least partly, by the decreasing capacity to remove dietary cysteine from the oxidative environment of the blood. Thiol antioxidants were found to ameliorate various aging-related processes but obviously ought to be used with caution in consideration of the oxidative environment of the blood. PMID- 12559404 TI - Dissociation between functional senescence and oxidative stress resistance in Drosophila. AB - Many studies strongly suggest a causal link between oxidative stress and determination of life span. The relationship between oxidative stress and age related functional declines, however, is less clear. Additionally, the full spectrum of functional declines associated with aging has not been systematically evaluated in the fruit fly, Drosophila melanogaster, one of the leading models for aging research. Toward a more comprehensive assessment of functional senescence in Drosophila, we evaluated a series of behaviors in control flies of increasing ages. Our studies reveal a novel age-dependent functional decline in the olfactory system and confirm previous reports of age-related locomotor defects in flies. Behavioral responses to electric shock and light are maintained in aged flies. Thus, some sensory systems senesce during the first several weeks of life while others do not. Interestingly, the age-dependent functional declines in olfactory and locomotor systems are indistinguishable in control flies and methuselah, a mutant with enhanced resistance to oxidative stress and increased life span. Our results indicate that enhanced resistance to oxidative stress and extension of life span do not necessarily confer protection from age-related functional declines. PMID- 12559405 TI - No reduction of metabolic rate in food restricted Caenorhabditis elegans. AB - Dietary restriction (DR) is the most consistent means of extending life span throughout the animal kingdom. Multiple mechanisms by which DR may act have been proposed but none are clearly predominant. We asked whether metabolic rate and stress resistance is altered in Caenorhabditis elegans in response to DR. DR was imposed in two complementary ways: by growing wild-type worms in liquid medium supplemented with reduced concentrations of bacteria and by using eat-2 mutants, which have a feeding defect. Metabolic rate was not reduced when we fed wild-type worms reduced food and was up-regulated in the eat-2 mutants in liquid culture, as assessed by oxygen consumption rate and heat production. The specific activity levels of the antioxidant enzymes superoxide dismutase (SOD) and catalase showed small increases when we reduced food in wild-type worms, but restricted worms acquired no elevated protection against paraquat and hydrogen peroxide. eat-2 mutants showed elevated specific activities of SOD and catalase relative to wild type in liquid culture. These results indicate that the effects imparted by DR and the eat-2 mutation are not identical, and they contradict, at least in C. elegans, the widespread belief that CR acts by lowering the rate of metabolism. PMID- 12559406 TI - Axenic growth up-regulates mass-specific metabolic rate, stress resistance, and extends life span in Caenorhabditis elegans. AB - Culture in axenic medium causes two-fold increases in the length of development and adult life span in Caenorhabditis elegans. We asked whether axenic medium imposes dietary restriction (ADR), and causes changes in metabolic activity and stress resistance. Eat mutants, which have a reduced food intake, were studied in parallel with wild-type worms to assess potential synergistic actions of axenic culture and food restriction. We found that axenic culture enhances metabolic activity as assessed by mass-specific oxygen consumption rate and heat production. Axenic culture also caused higher activities of the antioxidant enzymes superoxide dismutase and catalase, and led to increased resistance to high temperature, which was further exacerbated by mutation in eat-2. These results show that axenic medium up-regulates a variety of somatic maintenance functions including oxidative and thermal stress resistance and that food restriction due to axenic growth and to mutation in eat-2 are very similar but not identical. PMID- 12559407 TI - Modulation of cell-phenotype during in vitro aging. Glycosaminoglycan biosynthesis by skin fibroblasts and corneal keratocytes. AB - The aim of this study was to compare keratocyte and fibroblast phenotypes during in vitro aging by comparing their biosynthesis of glycosaminoglycans using explant and cell cultures. Human skin and corneal explant cultures were realised with Dulbecco Modified Eagle's medium containing 3H glucosamine. Sequential cell cultures were studied at different passages for GAGs biosynthesis by 3H glucosamine incorporation followed by selective degradation with specific hydrolases. Radioactivity was determined and each GAG fraction evaluated. KS and DS are the major components synthesised by corneal explant culture. During in vitro aging, keratocytes synthesised 41% less KS between passages 4-9 with a decrease by 26% of the proportion of DS observed in the same conditions. In skin explant cultures, as expected the major components are CS and hyaluronan (HA). In the first cell passage studied compared with skin organ cultures we could notice a strong decrease of the proportions of DS and KS compensated by an increase of the proportion of HA. During the successive passages of fibroblasts, the proportions of DS and HS decreased (-30 and -62%, respectively) and those of KS increased (+90%). These results indicate that there remain measurable differences between keratocyte and fibroblast phenotypes as far as GAG-synthesis is concerned all though the successive passages, starting from explant cultures and up to the limits of in vitro cell passages. PMID- 12559408 TI - Mitotic brain cells are just as prone to mitochondrial deletions as neurons: a large-scale single-cell PCR study of the human caudate nucleus. AB - Mitochondria are considered a key element in the process of organismic aging, because of their fundamental role in cellular energy generation. In the course of oxidative phosphorylation, harmful free radicals are continuously produced damaging the mitochondrial (mt) genome. One of the consequences is the occurrence of large-scale deletions in mtDNA molecules. The 4977 bp common deletion accumulates exponentially with age, in a mosaic pattern, especially in postmitotic tissues. In order to investigate whether certain cell characteristics underlie this pattern of distribution, and to look for possible age-related changes, two cell types in the caudate nucleus of the human brain from five young and five senescent subjects were analysed by single-cell PCR.MAP2-positive neurons and GFAP-positive astrocytes were isolated by micromanipulation. For each of the 10 cases, at least 30 cells of each type were collected and subjected to PCR individually. Screening for the presence of the common deletion yielded no significant differences in relative distribution, neither between astrocytes and neurons, nor between healthy young and old humans. Our results imply that the age dependent increase of the common deletion cannot come about by an increase of independent deletion events in a greater proportion of cells, and that mitotic rate is not a major cellular risk factor for deletion accumulation in the caudate nucleus. PMID- 12559409 TI - Transgenic mice expressing the human C99 terminal fragment of betaAPP: effects on spatial learning, exploration, anxiety, and motor coordination. AB - The functional consequence of beta-amyloid precursor protein (betaAPP) manipulation on behavior was assessed in Tg13592 mice, characterized by transgene expression of the 99 amino acid C-terminal sequence of human betaAPP in brain and skeletal muscle but with plaque formation only in muscle. By comparison to the C57BL/6 background strain controlled for age and gender, Tg13592 transgenic mice had fewer movements in an automated chamber and fewer enclosed arm entries in the elevated plus-maze. This hypoactivity was probably due to a loss in the motivation to explore novel environmental stimuli rather than motor weakness or anxiety. In addition, the acquisition of place learning in the Morris water maze task was impaired in Tg13592 mice. The transgenic mice were not impaired in a probe trial or while swimming toward a visible platform. These results are concordant with the hypothesis that transgene expression of the C-terminal sequence of human betaAPP in brain is sufficient for causing behavioral abnormalities. The hypoactivity and the spatial learning deficit were associated with higher cytochrome oxidase activity seen in thalamic nuclei, indicating that altered regional brain metabolism caused by betaAPP transgene expression may be responsible for the behavioral changes. PMID- 12559410 TI - Met-enkephalin modulates lipid peroxidation and total sialic acid level in CBA mice in age- and sex-dependent manners. AB - Age- and sex-associated differences in lipid peroxidation (LPO), and total sialic acid content (TSA) in response to abuse of drugs have been reported both in humans and experimental animals. However, no data on the influence of gender and age on these parameters have been reported for methionine-enkephalin (MENK). In this study we examined the influence of age and gender on MENK-induced LPO levels in the liver and TSA content in splenocytes of CBA mice. LPO production, which was age- and gender-associated was differentially regulated by MENK at a dose of 10 mg or 2.5 mg/kg body weight. At the higher dose, MENK stimulated LPO production in younger males and females but suppressed only in older male mice. At the lower dose, MENK induced strong suppression in males while being without any effect in females. In TSA levels, the age-associated increase was greater in males and much lower in females, with higher TSA levels in younger (2.5, 4.5 months) and decreased levels in older female mice (9 months) being observed. Contrary to the effect on LPO level, TSA level in MENK-treated mice was suppressed in both sexes but only in young 2.5-month-old mice. These data provide evidence that some immunomodulatory properties of MENK are age- and gender associated which may be relevant to the potential use of MENK as adjuvant therapy in patients with immunocompromised status. PMID- 12559412 TI - Old as mature LOU/c/jall rats enhance protein selection in response to a protein deprivation. AB - Previous experiments have shown a strong decrease in protein consumption as rats grow old. This result could be induced by an impaired regulation of feeding behaviour. Present study investigated the ability of ageing rats to adapt their protein intake to protein requirements. Four groups of self-selecting LOU/c/jall male and female rats (4, 16, 22, 28 months at the beginning of the experiments) were submitted to 4-days of protein deprivation periods while their macronutrient intakes were monitored. Moreover, they were submitted to a 4-day fasting period. After reintroduction of proteins, old and senescent rats were able to increase their protein intake in response to the specific protein need created by the protein deprivation. They were also able to increase caloric intake after the fasting period. These results led to the conclusion that the loss of appetite for protein observed with ageing reflects rather an adaptation than an impairment of the regulation of macronutrient choice. Another important observation was that older animals did not recover initial body weight after a body weight loss even if they ate as many calories as younger animals. This data supports a decrease in the efficacious utilization of body fuels in old and senescent rats. PMID- 12559411 TI - Effect of aging and late onset dietary restriction on antioxidant enzymes and proteasome activities, and protein carbonylation of rat skeletal muscle and tendon. AB - Many studies have shown that lifelong dietary restriction (DR) can retard aging processes. Very few reports, however, are found that examined the effect of late onset DR on biochemical parameters in aging animals [Goto, S., Takahashi, R., Araki, S., Nakamoto, H., 2002b. Dietary restriction initiated in late adulthood can reverse age-related alterations of protein and protein metabolism. Ann. NY Acad. Sci. 959, 50-56]. We studied the effect of every-other-day feeding, initiated at the age of 26.5 months and continued for 3.5 months, on antioxidant enzymes, protein carbonyls, and proteasomes of the gastrocnemius muscle and tendon in rats. Age-related increase in the activity and content of Cu, Zn-SOD and the content of Mn-SOD was attenuated by the DR in both tissues. The same was true for glutathione peroxidase and catalase activities. Significant increase with age in protein reactive carbonyl derivatives (RCD) in the tendon was noted that was partially reversed by the DR. No significant change of RCD, however, was observed in the skeletal muscle. The age-related and DR-induced changes of the RCD in the tendon appeared to be associated with proteasome activity that decreases with age and increases by the DR. It is suggested that the late onset DR can have beneficial effects on the locomotive functions by reducing age associated potentially detrimental oxidative protein damage in the tendon. PMID- 12559413 TI - Influence of age on circadian rhythms of adrenoceptors in brain cortex, heart and submandibular glands of BALB/c mice: when circadian studies are not only useful but necessary. AB - In the present paper we study circadian rhythm variation of alpha1-adrenoceptors (alpha(1)ARs) and beta-adrenoceptors (betaARs) characteristics during aging. Experiments have been performed on mice's whole brain cortex, heart and submandibular glands, which have physiologically relevant regulatory functions at different ages. We used 36 young (4 months) and 36 old (26 months) mice; six animals from each group were sacrificed every 4h, starting at 8.00 a.m. Both the alpha(1)ARs and betaARs density in heart and brain and the betaARs of submandibular glands are significantly lower in old mice with respect to the young ones. The difference, however, if the comparison is performed only at a particular time, for example 8.00 a.m., is not always significant. The alpha(1)ARs density in submandibular glands does not show significant difference with aging, but opposite phases are observed. This study demonstrates that in dynamical systems such as receptors, single point analysis is not so suitable for a correct estimate of adrenoceptors density variation between young and old mice. PMID- 12559414 TI - Age-related alterations in the protein expression profile of C57BL/6J mouse pituitaries. AB - The aim of our study was to monitor the protein expression profile in pituitary glands of healthy C57BL/6J mice during aging. Pituitary glands of 4-week old (immature), 3-month old (mature), and >25-month old mice were analysed by proteomic tools such as two-dimensional electrophoresis and N-terminal micro sequencing. A change was detected in the expression of growth hormone after sexual maturation. Our particular interest, however, was directed against up regulated proteins in the old pituitary glands, which are proposed to be involved in the process of neuroendocrine aging. Among these proteins, the expression of glutathione-S-transferase (GST) and apolipoprotein A-1 were increased in old pituitaries. Furthermore, ubiquitin carboxyl-terminal hydrolase (UCH-L1) was significantly up-regulated in senescent C57BL/6J mouse pituitaries. Since only the rat homologue was known, we isolated and analysed the mouse UCH-L1 sequence. Since GST is involved in antioxidative defence and UCH-L1 is part of the ubiquitin/proteasome system, which is responsible for the removal of damaged proteins, these results suggest increased oxidative burden and an increased activity of the ubiquitin system. PMID- 12559415 TI - The expression of the RLF/INSL3 gene is reduced in Leydig cells of the aging rat testis. AB - The relaxin-like factor (RLF), which is the product of the INSL3 gene, is highly expressed in the fetal and adult-type Leydig cells of all species so far examined. In adult testes it is upregulated at puberty but appears subsequently to be expressed in a constitutive manner, independently of acute changes in the hypothalamic-pituitary-gonadal (HPG) axis. Functional hypogonadism with decreased testosterone is prevalent in the aging male. In order to test whether this is a property of the HPG axis, or of the Leydig cells themselves, RLF/INSL3 was used as an independent marker to assess rat Leydig cell differentiation status. Hybridization analysis showed that in the testes of old (2 years) rats, RLF/INSL3 mRNA expression was dramatically reduced, compared to young (3 months) animals. This was also evident at the protein level using immunohistochemistry. The results suggest that increasing functional hypogonadism in older male mammals is likely caused by a dedifferentiation of the Leydig cells themselves. PMID- 12559416 TI - Incidence of apoptosis increases with age in colorectal cancer. AB - The incidence of cancer increases with advancing age, but the biological behavior of cancer is known to be less aggressive in elderly people. Thus, the proliferative activity and extent of apoptosis of cancer cells were assessed in samples from 163 cases of colorectal cancer focusing on the age of patients, using Ki-67 labeling index (LI) and apoptotic index (AI) by terminal deoxynucleotidyl transferase (TdT)-mediated d-UTP nick end labeling method and staining for activated caspase-3. The Ki-67 LI of colorectal cancer ranged from 2.33 to 80.4% (mean 32.2%), while the AI ranged from 0.00 to 14.8% (mean 3.57%). Concerning the aging effect, linear and positive correlations were found for the Ki-67 LI of cancer with age (p<0.05) and the AI of cancer with age (p<0.05). However, in normal colorectal mucosa, aging of patients revealed a significant correlation only with the AI but not with the Ki-67 LI. The AI in earlier stages of cancers (stages 0 and 1) revealed a significant difference between younger cases (age<65) and more elderly cases (age>/=65) (p<0.05), however, the Ki-67 LI did not exhibit a significant difference. Therefore, an increased frequency of apoptosis in colorectal cancer tissues, especially in the earlier stages, may possibly explain the slower growth of colorectal cancers in the elderly. Next, the expressions of several regulatory molecules for the proliferation/apoptosis of tumor cells were determined. The results demonstrated a tendency for stronger and more frequent expressions of c-myc, Bak and Bax despite a rather weaker expression of Bcl-2 in cancer tissues from the elderly compared with those from the younger patients. The potential roles of these regulatory molecules on age change in the proliferation/apoptosis of colorectal cancers are discussed. PMID- 12559417 TI - Effect of age and hypoxia/reoxygenation on mRNA expression of antioxidative enzymes in rat liver and kidneys. AB - The influence of a short-time isobaric hypoxia as well as reoxygenation on markers of oxidative stress (MDA, total SOD, GSH) and on the mRNA expression of the antioxidative enzymes (Cu/Zn-and Mn-SOD, catalase, GSH reductase and GSH peroxidase) has been studied in liver and kidneys of young (6 months) and old (22 25 months) Wistar rats. In livers of old animals, the concentration of GSH, the activity of SOD, and the mRNA expression of the antioxidative enzymes (except Mn SOD) points to a restricted protection against oxidative stress or a lower production of ROS compared to young animals. Hypoxia resulted in a significant decrease of enzyme gene expression in both age groups. Reoxygenation caused an increase in mRNA of Cu/Zn-SOD and GPX in livers of young and of Mn-SOD in livers of old animals. In kidneys, gene expression of Cu/Zn-SOD, GSH reductase, and GPX was significantly higher in old animals compared to young animals. Whereas hypoxia caused a decrease of gene expression in the livers, it lead to a significant increase of Cu/Zn-SOD, catalase, and GSH reductase mRNA in kidneys of young rats. A reduced gene expression was observed after reoxygenation. In old kidneys, the expression of all enzymes except for catalase progressively declined in the hypoxic and reoxygenation groups. These data show that gene expression of antioxidative enzymes is affected by age and significantly differs between liver and kidney. PMID- 12559418 TI - Neurochemical changes related to ageing in the rat brain and the effect of DL alpha-lipoic acid. AB - Age-related impairments of cognitive and motor function have been linked to a number of deleterious morphological and functional changes involving different areas of the brain. Loss of neurotransmitters, their receptors and responsiveness to neurotransmitters are key manifestations of neurological ageing and age related disorders. In the present investigation we have evaluated the effect of DL-alpha-lipoic acid on neurotransmitters in discrete brain regions of young and aged rats. The levels of neurotransmitters were found to be lowered in aged rats. Moreover, DL-alpha-lipoic acid treated aged rats showed a increase in the status of dopamine, serotonin and norepinephrine. The results of this study provide evidence that DL-alpha-lipoic acid (a potent antioxidant) treatment can improve neurotransmitters during ageing. Hence, it can be concluded that DL-alpha-lipoic acid act as a potent neuromodulator in the brain of aged rats. PMID- 12559419 TI - Single molecule fluorescence and force microscopy. AB - The investigation of biomolecules has entered a new age since the development of methodologies capable of studies at the level of single molecules. In biology, most molecules show a complex dynamical behavior, with individual motions and transitions between different states occurring highly correlated in space and time within an arrangement of various elements. Recent advances in the development of new microscopy techniques with sensitivity at the single molecule have gained access to essentially new types of information obtainable from imaging biomolecular samples. These methodologies are described here in terms of their applicability to the in vivo detection and visualization of molecular processes on surfaces, membranes, and cells. First examples of single molecule microscopy on cell membranes revealed new basic insight into the lateral organization of the plasma membrane, providing the captivating perspective of an ultra-sensitive methodology as a general tool to study local processes and heterogeneities in living cells. PMID- 12559420 TI - Distribution of satellite cells in the human vastus lateralis muscle during aging. PMID- 12559421 TI - An optimised method to determine the degree of acetylation of chitin and chitosan by FTIR spectroscopy. AB - Linking up the user-friendly and low-priced FTIR with the more sophisticated and high-priced 13C CP/MAS NMR spectroscopies, an improved method to determine the degree of acetylation (DA) of chitins and chitosans was outlined. The method was established for the most complex polymorphic form (alpha-chitin) and for the most problematic range of DA values (most acetylated samples) and can easily be extended to the other polymorphic forms (beta- and gamma-chitins) and to other ranges of DA values. PMID- 12559422 TI - Structural and rheological properties of polysaccharides from mango (Mangifera indica L.) pulp. AB - The structure and rheological properties of water-soluble polysaccharides from industrialized mango pulp were investigated. Soluble fraction (SF) 2 was heterogeneous on high performance size exclusion chromatography, giving two peaks as determined by multi-angle laser light scattering and refractive index detectors. The presence of starch in SF2 was demonstrated by a positive iodine reaction and by 13C nuclear magnetic resonance (NMR) spectroscopy. The presence of pectic polysaccharides was shown by a calorimetric method, 13C-NMR spectroscopy and carboxyl reduction. The main pectic polysaccharide was polygalacturonic acid; type I rhamnogalacturonan was also detected. Analysis of the rheological properties of SF2 showed a pseudoplastic behavior up to 3 g x l( 1). 'Creep and recovery' tests and analysis performed under a dynamic state revealed a weak gel character for solutions at concentrations of 15, 20 and 30 g x l(-1). PMID- 12559423 TI - Optical characterization of armadillo acyl-CoA binding protein. AB - Acyl-CoA binding protein (ACBP) and fatty acid binding protein (FABP) are intracellular transporters of activated and free fatty acids, respectively. Unlike other tissues with active lipid metabolism, armadillo Harderian gland contains much more ACBP than FABP. To characterize armadillo ACBP structure and binding properties, we produced it in Escherichia coli and carried out detailed fluorescence and circular dichroism spectroscopy studies. The K(D) for palmitoyl CoA, measured directly by fluorescence and rotatory power, was 34+/-12 and 75+/ 39 nM, respectively. The structure of armadillo ACBP appears to be very similar to that of bovine and rat liver ACBPs. PMID- 12559424 TI - Preparation and characterization of molecular weight fractions of guar galactomannans using acid and enzymatic hydrolysis. AB - A procedure is described for the preparation of large amounts of guar galactomannan by acid hydrolysis that yields samples of various molecular weights (MW) with uniform polydispersity. This contrasts with preparation by enzymatic degradation that yields samples with a marked increase in polydispersity and a much broader molecular weight distribution (MWD). Acid hydrolyzed guar samples had a Mark-Houwink-Sakurada (MHS) relationship of [eta]=3.04x10(-4) M(w)(0.747) dl/g and a characteristic ratio of 11.87 as determined by gel permeation chromatography (GPC) and dilute solution viscometry. The Huggins coefficient for degraded guars is much smaller (approximately 0.4) than that of the native guar (approximately 0.79), suggesting a weakening of intermolecular association in guar prepared by acid hydrolysis. PMID- 12559425 TI - Rheology of protein gels synthesized through a combined enzymatic and heat treatment method. AB - Whey protein gels prepared under acidic conditions (pH<4.6) remain largely unutilized because of their weak and brittle nature in contrast to the favorable elastic gels produced at neutral or basic conditions. However, such usage is important, as low pH food products are desirable due to their shelf stability and less stringent sterilization processes. In this study, we use a two-step process involving enzyme followed by heat treatment to produce whey protein gels at low pH (4.0). Dynamic rheological measurements reveal that the gel elastic modulus and yield stress increase substantially when heat treatment is supplemented with enzyme treatment. Both the elastic modulus and yield stress increase with increasing enzyme concentration or treatment time. In contrast, the dynamic yield strain decreases with enzyme concentration but increases with time of enzyme treatment. These results are explained in terms of the enzyme treatment time affecting the diffusion of the enzyme within the gel. This in turn leads to two types of gel microstructure at short and long enzyme treatment times, with the extent of enzyme diffusion modulating the structure at intermediate times. PMID- 12559426 TI - Energetics of heparin binding to human acidic fibroblast growth factor. AB - The binding of low-molecular-weight heparin to an amino-terminal-truncated, 132 amino-acid, human acidic fibroblast growth factor form has been studied by isothermal titration calorimetry. This technique yields values for the enthalpy change and equilibrium constant, from which the Gibbs energy and entropy change are also calculated. Experiments in different buffers and pH values show that the protonic balance during the reaction is negligible. Experiments made at pH 7.0 with NaCl concentrations ranging from 0.20 to 0.60 M revealed changes in enthalpy and Gibbs energy in the range of -30- -17 and -27- -24 kJ x mol(-1), respectively. Isothermal titration calorimetry was also performed at different temperatures to obtain a value for the heat-capacity change at pH 7.0 and 0.4 M NaCl concentration of -96 J K- x mol(-1). A change in the length of heparin brought about no change in the thermodynamic parameters at 25 degrees C under the same experimental conditions. Changes upon ligand binding in the range of -50- 200 A2 in both polar and non-polar solvent-accessible surface areas were calculated from thermodynamic data by using different parametric equations taken from the literature. These values suggest a negligible overall conformational change in the protein when it binds to heparin and no formation of any protein protein interface. PMID- 12559427 TI - Chemical modification, characterization and structure-anticoagulant activity relationships of Chinese lacquer polysaccharides. AB - A natural lacquer polysaccharide with complex branches was separated into two fractions, LPH (MW 16.9x10(4)) and LPL (MW 6.85x10(4)). Results of 13C NMR and FT IR indicated they had the same structure. The treatment of LPL with sodium periodate led to a partial cut-off of side chains with 4-O-methyl-D-glucuronic acid in the terminal. These polysaccharides were sulfated in the presence of Py*SO3/DMSO. Depending on the reaction conditions, the products showed a different degree of sulfation (DS) ranging from 0.57 to 1.57 and different molecular weights ranging from 1.71x10(4) to 3.49x10(4). FT-IR analysis showed the equatorial primary OH at O-6 and the axial secondary OH at O-4 were sulfated. Activated partial thromboplastin time (APTT), prothrombin time and thrombin time (TT) assays showed the sulfated polysaccharides could prolong APTT and TT, but not TP. These activities strongly depended on the DS, the molecular weights (MW) and the branching structure of polysaccharides. DS of above 0.8 was essential for anticoagulant activity. The anticoagulant activity increased with the DS and the molecular weights. The molecular weights played a more important role. The branching structure of polysaccharides increased the activities. In our studies, the sulfated polysaccharides with the DS of 1.15 and the highest MW of 3.49x10(4) had the best blood anticoagulant activities. PMID- 12559428 TI - Synthesis of PHB by recombinant E. coli harboring an approximately 5 kb genomic DNA fragment from Streptomyces aureofaciens NRRL 2209. AB - An approximately 5.0 kb Sau3A I genomic DNA fragment from Streptomyces aureofaciens NRRL 2209 was cloned in a plasmid vector and introduced into Escherichia coli. The recombinant E. coli accumulated polyhydroxyalkanoates (PHAs) as cytoplasmic inclusions. The accumulated PHA was identified as the isotactic homopolymer of PHB with a molecular weight of 2.85x10(5). Purified PHB granules were spherical with an average size of 1.1 microm and of stable configuration. DSC thermogram suggested high crystalline nature of the polymer. Maximum thermal degradation of the biopolymer occurred between 250 and 340 degrees C. Recombinant E. coli cells preferentially utilized glycerol as the carbon source and accumulated 25-28 times more PHB than the native S. aureofaciens. PMID- 12559429 TI - Effect of alpha2M on earthworm fibrinolytic enzyme III-1 from Lumbricus rubellus. AB - Though it is known that human alpha2-macroglobulin (alpha2M) inhibits most proteases, the effect of alpha2M has not been investigated on earthworm fibrinolytic enzymes (EFEs) from Lumbricus rubellus, which could be transported from intestine epithelium into blood as an intact molecule (Fan et al., Biochim. Biophys. Acta 1526 (2001) 286). The activity of earthworm fibrinolytic III-1 (EFE III-1) decreased to 65% when incubated with alpha2M, while it decreased to 30% in plasma under the same conditions. The first order rate of the inactivation of EFE III-1 with alpha2M was similar to that of fast phase with plasma, indicating that alpha2M may be the inhibitor initially binding to the enzyme in blood. SDS-PAGE showed that incubation of EFE-III-1 with alpha2M a released fragment ( approximately 90 kDa), followed by formation of a high molecular weight complex (approximately 700 kDa). There was a linear relationship between the apparent inhibition rate constant (k1) and [alpha2M], by double reciprocal plot. It was suggested, as described by Tsou (Acta Biochem. Biophys. Sinica 5 (1965) 398) and Tian (Biochem. J. 21 (1982) 1028), that the mechanism of alpha2M/EFE-III-1 interaction could be coincided with a complexing irreversible inhibition. Experiments in both the inactivation and the intrinsic fluorescence showed that alpha2M bound to the enzyme mole by mole equivalently. The intrinsic fluorescence of alpha2M was enhanced with an observable blue shift in emission maxima, suggesting that alpha2M was one of the important inhibitors to EFEs when it absorbed into blood. PMID- 12559430 TI - Organisation of the external region of the starch granule as determined by infrared spectroscopy. AB - Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) was used to study the external regions of starch granules. Native starches (wheat, potato, maize, waxy maize and amylomaize) were analysed and compared to gelatinised and acid-hydrolysed starches. The IR spectra of potato and amylomaize starches were closer to that of highly ordered acid-hydrolysed starch than the other starches. FTIR was not able to differentiate between A- and B-type crystallinity so the difference observed between starches was not related to this factor. The variation between starch varieties was interpreted in terms of the level of ordered structure present on the edge of starch granules with potato and amylomaize being more ordered on their outer regions. This could explain the high resistance of both these starches to enzyme hydrolysis. PMID- 12559432 TI - Inactivation and conformational changes of lactate dehydrogenase from porcine heart in sodium dodecyl sulfate solutions. AB - The inactivation and conformational changes of porcine heart lactate dehydrogenase (LDH) have been studied in sodium dodecyl sulfate (SDS) solutions. Increasing SDS concentration led to a quick and concentration-dependent inhibition of the enzyme, with complete inactivation within 5 min in the presence of 1.0 mM SDS. Meanwhile, fluorescence emission and circular dichroism spectra were used to follow the conformational changes of the enzyme during this process, concurrently showing that SDS less than 1.0 mM induced only limited conformational changes to LDH. The above results are in accordance with the suggestion by Tsou (Trends Biochem. Sci. 11 (1986) 427; Science 262 (1993) 380) that the active site usually be more flexible than the enzyme molecule as a whole. Furthermore, the results of polyacrylamide gel electrophoresis (PAGE) implied that unfolding intermediates were presented in the above process. When the SDS concentration used to treat LDH was increased, the bands of native enzyme on native PAGE faded and finally almost disappeared. Meanwhile, multiple bands with lower mobility but no activity emerged behind and enhanced correspondingly. Fast protein liquid chromatography indicated that dissociation occurred during the course of denaturation. The reasons for the above phenomena have been discussed. It was suggested that SDS, binding to LDH to form different LDH-SDS complexes, conferred an array of different unfolding states over the enzyme, and in turn resulted in the formation of the multiple bands on the native PAGE. PMID- 12559431 TI - Isolation, chemical investigation and antiviral activity of polysaccharides from Gracilaria corticata (Gracilariaceae, Rhodophyta). AB - Polysaccharides were sequentially extracted from the agarophyte Gracilaria corticata. Chemical analysis combined with infrared spectroscopy showed that the cold water extracted material consists mainly of a high molecular weight sulfated galactan. Most of the sulfate groups are alkali labile and are located at C-4 of the 1,3-linked D-galactose units and C-6 of the 1,4-linked L-galactose residues. The autoclaved extracts contain agar type polysaccharide having a high pyruvate content and a variable degree of methylation, but were contaminated with floridean starch. 1H-NMR studies indicate that methoxyl groups, when present, occur at C-6 of the 1,3-linked D-galactose units and C-2 of the 1,4-linked L galactose residues of agar polymer. Bioassays showed that a high molecular weight galactan sulfate, exhibited selective antiviral activity against herpes simplex virus types 1 and 2, likely due to an inhibition of the initial virus attachment to the host cell. PMID- 12559433 TI - On the dissection of the unfolding reaction by the dissolution thermodynamics of N-alkyl amides. AB - Dissection of the unfolding thermodynamics based on small molecule dissolution led to controversial interpretations. It is proposed here that uncertainty about water transfer processes to be used in first approximation analyses of protein stability may be removed (i) separating liquid-dissolution-like effects from solid-like packing contributions; (ii) taking into account both peptide and side chain dissolution; and (iii) analysing the water-dependent part of the denaturation reaction by the dissolution thermodynamics of liquid N-alkyl amides. Based on these criteria, this paper analyses the entropy of the aqueous transfer of liquid N-alkyl amides filling a gap in a recent model of the unfolding energetics, which was limited to the enthalpy. Both enthalpic and entropic changes accompanying the liquid-dissolution-like immersion of internal amino acid residues in water during unfolding may be unambiguously described within this context. Although the model developed does not deepen our knowledge of protein unfolding, it may be of help in the analysis of whether liquid-dissolution-like effects or solid-like packing contributions play the major role in determining protein stability at elevated temperatures. PMID- 12559434 TI - Enzymic preparation of water-soluble chitosan and their antitumor activity. AB - Water-soluble low-molecular-weight (LMW) chitosan was prepared from enzymatic hydrolysis with efficient hemicellulase. The hydrolysates were separated by ultrafiltration membranes. A separated fraction with Mw more than 5x10(3) and with a degree of deacetylation of 58% was water-soluble in the free amine form. The intraperitoneal injection of LMW chitosan and its N-acetyl product inhibited the growth of sacroma 180 (S180) tumor cells in the mice, and the maximum inhibitory rate reached 64.2%. The oral administration was also effective on decreasing weight of tumor, and the maximum inhibitory rate reached 33.7%. The Water-soluble chitosan with higher Mw than hexamer might have better antitumor activity. PMID- 12559436 TI - Evidence for a central role for selective mesenteric angiography in the management of the major vascular complications of pancreatitis. AB - BACKGROUND: Although mesenteric angiography and embolization are established radiologic procedures, the evidence-base to aid decision making regarding selection of these procedures in the emergent situation in patients with hemorrhagic complications of pancreatitis is limited. METHODS: A retrospective analysis of 19 patients with hemorrhagic complications of pancreatic inflammatory disease (acute pancreatitis, chronic pancreatitis, and pseudocyst) referred over a 4-year period and identified at the point of mesenteric angiography in order to determine the influence of interventional radiologic maneuvers on outcome. RESULTS: Mesenteric angiography localized bleeding in 15 (79%), with 11 (58%) embolizations undertaken. There was one (9%) procedure-related complication and 3 (27%) rebleeds. Of 11 patients undergoing embolization, 8 (73%) required no further intervention for bleeding and 8 (73%) survived. CONCLUSIONS: Mesenteric angiography is valuable in localizing bleeding in patients with major vascular complications of pancreatic inflammatory disease. Angiographic embolization can achieve definitive hemostasis and stabilize a critically ill patient to permit disease reappraisal. PMID- 12559435 TI - A long-term cohort study of outcome after cholecystectomy for chronic acalculous cholecystitis. AB - BACKGROUND: Cholecystectomy is effective therapy for chronic calculous cholecystitis (CCC). The long-term outcome of patients treated with cholecystectomy for chronic acalculous cholecystitis (CAC) is unknown. METHODS: A controlled, retrospective cohort study assessing biliary pain (preoperative and at follow-up) in postcholecystectomy patients with CAC or CCC was performed. RESULTS: In 19 CAC and with matched CCC control patients, the mean duration of symptoms before surgery was 38.3 months (95% CI, 16.4 to 60.2) and 8.1 months (95% CI, 5.4 to 10.8), respectively. The mean follow-up for both groups was 8.37 +/- 1.13 years. Both groups benefited from cholecystectomy (P <0.001), and both were equally likely to be pain-free upon long-term follow-up (95% CAC versus 84% CCC, P >0.05). CONCLUSIONS: There was no difference in outcome between the groups after an average follow-up of 8.37 years. Postcholecystectomy patients with chronic cholecystitis and no gallstones have long-term, complete pain resolution, similar to patients with gallstones. PMID- 12559437 TI - Nonobstructive dysphagia and recovery of motor disorder after antireflux surgery. AB - BACKGROUND: Changes in motor disorder after Nissen 360 degrees surgery were studied based on clinical signs of preoperative nonobstructive dysphagia. MATERIALS AND METHODS: Forty-seven patients undergoing Nissen 360 degrees fundoplication for gastroesophageal reflux were studied with pH recording and esophageal manometry before and 1 year after fundoplication. Amplitude of contraction of the distal third of the esophagus (ACDTE) and the presence of primary propulsive waves were studied. RESULTS: Fourteen patients had clinical signs of preoperative dysphagia. Of these, 50% had an ACDTE lower than 30 mm Hg, and 71.4% nonpropulsive waves (P <0.05). Forty-three percent and 30%, respectively, of patients with dysphagia recovered ACDTE and the presence of primary propulsive waves 1 year after the procedure, as compared with 66.6% (P <0.05) and 81.8% (P <0.01%) of patients without dysphagia. CONCLUSIONS: A correlation was found between preoperative dysphagia and esophageal motility disorders (P <0.05). One year after fundoplication, recovery was significantly higher in patients without preoperative dysphagia. PMID- 12559438 TI - Description of M-shaped preperitoneal hernioplasty for inguinocrural hernias. AB - BACKGROUND: The efficacy of preperitoneal herniorrhaphy is controversial in the surgical literature. The development of totally extraperitoneal video-assisted hernioplasty has stimulated further interest in long-term results of open techniques. Some of the main advantages of the two most extended preperitoneal open approaches (Stoppa and Nyhus operations) are combined in the author's technique described here. METHODS: Between March 1995 and June 1998, 180 consecutive patients with a total of 202 inguinocrural hernias were treated by an open sutureless preperitoneal repair, using a lateral approach, with placement of a 12 x 10 cm polypropylene mesh. RESULTS: Mean operating time was 48 minutes. Overall morbidity rate was 7%. Mean hospital length of stay was 2.16 days. After an average follow up of 3.5 years, the overall recurrence rate was 0.57%. CONCLUSIONS: M-shaped preperitoneal hernioplasty is a single, safe, and easy repair for inguinocrural hernias, regardless of the type of hernia or the concomitant clinicopathological features of the patient. PMID- 12559439 TI - Sentinel lymph node biopsy followed by delayed mastectomy and reconstruction. AB - BACKGROUND: The role of sentinel lymph node (SLN) biopsy with total mastectomy is evolving. In patients who desire mastectomy with immediate reconstruction, the final pathologic results of the SLN may create unique problems. Specifically, if the SLN is found to be positive on final pathology, the reconstructed patient would generally require a potentially difficult re-operation on the remaining axillary nodes. The purpose of this study was to review the results of patients who underwent an initial SNL biopsy followed by a planned mastectomy and reconstruction. METHODS: A chart review of patients who underwent staged SLN biopsy with subsequent definitive procedure between 1997 and 2001 was conducted. These were evaluated with regard to type of tumor, status of sentinel node, and design of subsequent operation. RESULTS: There were 40 patients who underwent an initial SLN biopsy followed by a staged mastectomy with reconstruction. Tumors included high-grade carcinoma in situ (n = 4), infiltrating ductal carcinoma (n = 28), invasive lobular carcinoma (n = 4), mucinous carcinoma (n = 1), adenoid cystic carcinoma (n = 1), and mixed ductal and lobular carcinoma (n = 2). Tissue biopsy was obtained by either open (n = 9) or needle (n = 31) technique. Twenty five patients had a negative SLN biopsy and a delayed total mastectomy with immediate reconstruction. Positive SLNs were identified in 15 patients (37%). Eight patients had macroscopic nodal metastases and underwent a delayed modified radical mastectomy and immediate reconstruction. Seven patients had microscopic nodal metastases and 3 declined further axillary dissection. They proceeded with total mastectomy and immediate reconstruction. CONCLUSIONS: These data suggest that a substantial proportion of patients treated with SLN biopsy, simple mastectomy, and reconstruction will have positive sentinel lymph nodes. Thus, the ideal approach for patients who wish to have reconstruction should involve an initial SLN biopsy as a separate procedure. If the SLN is benign, the patient may undergo a total mastectomy with immediate reconstruction. However, a patient with a positive SLN may proceed to a modified radical mastectomy with immediate reconstruction. This treatment algorithm eliminates a potentially difficult reoperation on the axilla following reconstruction. PMID- 12559440 TI - Outcomes of sentinel node biopsy for breast cancer in British Columbia, 1996 to 2001. AB - BACKGROUND: This study evaluated the outcomes of the first 5 years of sentinel node biopsy (SNB) in British Columbia (BC), Canada, 1996 to 2001. METHODS: There were 547 SNB procedures for breast cancer performed by 29 surgeons at 12 hospitals in BC between October 1996 and July 2001. Identification, accuracy, and false-negative rates were determined and correlated to patient, tumor, and surgical factors with the chi-square test. RESULTS: SNB mapping was performed using blue dye alone (15%), radiopharmaceutical alone (6%), or both (79%). A completion axillary dissection was performed in 93%. A median of 2 (range 1 to 16) sentinel nodes was biopsied. The overall identification rate was 88%, accuracy was 92%, and false-negative rate was 22%. All rates were improved in younger (age <50 years) compared with older women. A positive lymphoscintiscan and the mapping agent used were associated with higher identification rates but not accuracy or false negative rates. Increasing surgeon experience was not significantly associated with improvements in identification or false-negative rates. CONCLUSIONS: The potential of SNB was not fully translated into surgical practice in BC by 2001. PMID- 12559441 TI - Motion restriction and axillary web syndrome after sentinel node biopsy and axillary clearance in breast cancer. AB - BACKGROUND: We evaluated the prevalence of motion restriction and axillary web syndrome (AWS) after sentinel node biopsy (SNB) and axillary clearance (AC) in a prospective study. AWS is a self-limiting cause of early postoperative morbidity after axillary surgery. Limited range of motion associated with palpable cords of tissue in the axilla are typical for AWS. METHODS: Altogether 85 breast cancer patients who underwent SNB only (49 patients) or SNB and AC (36 patients) were examined before and after surgery. The range of shoulder flexion and abduction and the presence of AWS were registered. RESULTS: The range of shoulder movements was restricted in 24 (45%) patients after SNB only and in 31 (86%) patients who also underwent AC (P = 0.002). AWS was encountered in 10 (20%) patients with SNB and in 26 (72%) with AC (P <0.00005). CONCLUSIONS: In the SNB group, significantly less early postoperative morbidity was observed. PMID- 12559442 TI - The role of cardiac morbidity in short- and long-term mortality in injured older patients who survive initial resuscitation. AB - BACKGROUND: Elderly patients are an increasingly larger group of injured trauma care patients. Comorbidities influence outcome. Little is known of short- and long-term mortality in the elderly who survive initial resuscitation. METHODS: Short- and long-term mortality was retrospectively analyzed in 363 consecutively injured patients (Injury severity score >15) surviving more than 3 days after admission to a level 1 trauma center (including 197 patients >60 years). Cardiac morbidity was the focus. RESULTS: Survival to hospital discharge was similar comparing older patients with the entire group. Mortality increased incrementally with age. In older patients, cardiac morbidity was observed in 28% (fatal in 7); 2-year mortality was 36% (older group) and 60% (patients sustaining cardiac complications). Most elderly (80%) were discharged to long-term care. CONCLUSIONS: Elderly who survive initial resuscitation are as likely to survive to discharge as younger patients, but long-term survival is significantly lower as age increases. Cardiac morbidity is associated with higher long-term mortality. Most elderly are discharged to long-term care. PMID- 12559443 TI - Aggressive resection is indicated for cecal diverticulitis. AB - BACKGROUND: Because of the difficulties in preoperative diagnosis and controversies in the management, cecal diverticulitis has received much discussion in the literature. There, however, are still many questions that remain unanswered. METHODS: During a 5-year period, 112 patients with a clinical diagnosis of cecal diverticulitis were treated. Twenty-seven patients were excluded because of uncertainty in diagnosis or incomplete data collection, leaving 85 patients as the study group. The diagnosis of cecal diverticulitis was made by pathology, surgical findings, or image study. RESULTS: Nonoperative management was applied to 18 patients initially. Three patients had recurrent diverticulitis during follow up. These patients responded satisfactorily to another course of medical treatment. Laparotomy was performed in 67 patients. Acute appendicitis was the preoperative diagnosis in 47 patients (70%). Of the other 20 patients, 6 received operation because of repeated attack of diverticulitis, 7 had preoperative computed tomography (CT) diagnosis of cecal diverticulitis with perforation, 5 had preoperative diagnosis of cecal tumor, and 2 had medical treatment failure. All these 20 patients received right hemicolectomy. In the 47 patients with a preoperative diagnosis of acute appendicitis, 24 received appendectomy, 9 received diverticulectomy, and 14 received right hemicolectomy. Overall, 34 patients received right hemicolectomy, 9 received diverticulectomy, and 24 received appendectomy only. In the right hemicolectomy group, there were 2 deaths with underlying diseases and 5 complications. In the appendectomy group, there was no postoperative mortality, but in 7 patients recurrent diverticulitis developed. Three of them required right hemicolectomy. CONCLUSIONS: The natural history of cecal diverticulitis varies from benign and self-limiting to fulminant in the oriental population. Less than 40% (32 of 85) of patients were successfully treated with conservative methods initially and had no recurrence during the follow-up period. We recommend aggressive surgical resection for patients with a definite diagnosis. Adjuvant appendectomy without resection of the lesion should be considered only in uncomplicated patients whose diagnosis is in doubt. PMID- 12559444 TI - Double gloving and practice attitudes among surgeons. AB - BACKGROUND: Exposure to blood and body fluids while operating places surgeons at risk. Double gloving is a safety measure, which decreases this risk. However, many surgeons do not incorporate this precaution into their personal practice. This study evaluates surgeons' gloving practices and hepatitis status. METHODS: A questionnaire was sent to all consultant surgeons and residents in the surgical specialties in the Capital Health region, University of Alberta. A double mail out design was used. A second questionnaire, containing information on safety issues, was sent to the general surgeons (consultants and residents) who did not double glove to ascertain whether this information would change their practice. RESULTS: In all, 268 surgeons and residents were sent the original questionnaire; 170 replied (63.4% response rate.) Fifty-seven percent of the respondents do not double glove (none of the urologists double glove versus 87% of orthopedic surgeons). The most common reason sited was a decrease in manual dexterity (46%). Ninety-seven percent of respondents are immunized for hepatitis B with 53% having had their titres recently checked. Thirty-seven general surgeons received the evidence on safety issues but only 9 (23%) of them would change their practice as a result of this information. CONCLUSIONS: The majority of surgeons and residents do not double glove. Even when provided with good evidence of efficacy, few surgeons contemplate adopting safety techniques. PMID- 12559445 TI - Learning curves and impact of previous operative experience on performance on a virtual reality simulator to test laparoscopic surgical skills. AB - BACKGROUND: The study was carried out to analyze the learning rate for laparoscopic skills on a virtual reality training system and to establish whether the simulator was able to differentiate between surgeons with different laparoscopic experience. METHODS: Forty-one surgeons were divided into three groups according to their experience in laparoscopic surgery: masters (group 1, performed more than 100 cholecystectomies), intermediates (group 2, between 15 and 80 cholecystectomies), and beginners (group 3, fewer than 10 cholecystectomies) were included in the study. The participants were tested on the Minimally Invasive Surgical Trainer-Virtual Reality (MIST-VR) 10 consecutive times within a 1-month period. Assessment of laparoscopic skills included time, errors, and economy of hand movement, measured by the simulator. RESULTS: The learning curves regarding time reached plateau after the second repetition for group 1, the fifth repetition for group 2, and the seventh repetition for group 3 (Friedman's tests P <0.05). Experienced surgeons did not improve their error or economy of movement scores (Friedman's tests, P >0.2) indicating the absence of a learning curve for these parameters. Group 2 error scores reached plateau after the first repetition, and group 3 after the fifth repetition. Group 2 improved their economy of movement score up to the third repetition and group 3 up to the sixth repetition (Friedman's tests, P <0.05). Experienced surgeons (group 1) demonstrated best performance parameters, followed by group 2 and group 3 (Mann Whitney test P <0.05). CONCLUSIONS: Different learning curves existed for surgeons with different laparoscopic background. The familiarization rate on the simulator was proportional to the operative experience of the surgeons. Experienced surgeons demonstrated best laparoscopic performance on the simulator, followed by those with intermediate experience and the beginners. These differences indicate that the scoring system of MIST-VR is sensitive and specific to measuring skills relevant for laparoscopic surgery. PMID- 12559446 TI - Improved quality of life with jejunal pouch reconstruction after total gastrectomy. AB - BACKGROUND: There is increasing evidence that the effect of jejunal pouch reconstruction is satisfactory for reservoir function in several randomized control studies. However, these studies were performed in patients with advanced gastric cancer, where significant numbers of the patients died of disease recurrence. In order to exclude the influence of disease recurrence, we performed jejunal pouch reconstruction after total gastrectomy in patients with early gastric cancer in a randomized controlled study and investigated whether or not an improved quality of life (QOL) was observed with jejunal pouch reconstruction. METHODS: Fifty consecutive patients receiving total gastrectomy for early gastric cancer were prospectively divided into the Roux-en-Y reconstruction group without pouch (RY group) or the jejunal pouch reconstruction group (pouch group). Body weight, eating capacity, QOL assessment by gastrointestinal symptom rating scale (GSRS), nutritional parameters, endoscopical examination, 24-hour pH monitoring and Bilitec monitoring were evaluated at 3, 12, and 48 months after surgery. RESULTS: Jejunal pouch reconstruction provided the better QOL than Roux-en-Y reconstruction without pouch both at short-term and long-term periods in a randomized control study. Moreover, as a new finding, pouch reconstruction provided less bile reflux into the esophagus compared with Roux-en-Y reconstruction. CONCLUSIONS: Jejunal pouch reconstruction provided improvement of QOL in patients receiving total gastrectomy. PMID- 12559447 TI - Additional right parasternal incision without thoracotomy provides alternative access for hepatic resection. AB - BACKGROUND: Although hepatic resections are performed with various access approaches according to the location of the tumor, the extent of resection and patient's physical constitution, the exposure of the root of the hepatic veins is not optimal in some patients by any conventional incision. METHODS: An additional 5 cm of right parasternal skin incision followed by division of two costal cartilages was performed in 8 patients, which demonstrated poor exposure of the upper part of the liver by abdominal incision. RESULTS: In all cases, satisfactory exposure of the operative area was obtained without thoracotomy, and hepatic resection procedure was carried out without significant events. CONCLUSIONS: This simple technique without necessitating rotation of the operating table may be useful as an alternative method of access to the liver especially when fine isolation around the root of the hepatic veins is required. PMID- 12559448 TI - An attempt to clarify indications for hepatectomy for liver metastases from breast cancer. AB - BACKGROUND: Liver metastases (LM) from breast cancer are generally considered as disseminated disease with a poor prognosis. However in selected patients hepatectomy may be an important adjunct to systemic treatment. METHODS: Fifty four breast cancer patients (mean age 49.2 +/- 5.2 years) with LM as the sole site of metastatic disease (except for bone metastases in 3 patients) underwent hepatectomy between 1986 and 2000. The mean number of LM was 4.0 +/- 8. All patients presented either a stable disease or an objective response to chemotherapy. The last 25 patients also underwent hepatic artery catheter installation in order to receive postoperative hepatic artery infusion chemotherapy (HAIC). RESULTS: The postoperative morbidity was 12.9%. There was no postoperative mortality. R0 and R1-R2 resections were obtained in, respectively, 81.5% and 18.5% of patients. After a median follow-up of 32 months the median survival was 34 +/- 9 months, with 3- and 5-year overall survival rates of 50% and 34%, and 3- and 5-year disease-free survival rates of 42% and 22%, respectively. The number of LM, the presence of hilar lymph nodes (33%), and the completeness of resection had no significant prognostic impact. The only factor influencing survival in both the univariate and multivariate analysis was the hormone receptor status (P = 0.03): the relative risk of death was increased by 3.5-fold when negative. In the HAIC group, the liver recurrence rate decreased from 60.5% to 31.2% without any impact on global survival. CONCLUSIONS: Hepatectomy is beneficial for selected patients with isolated LM. Indications should be based more on technical (low operative risk, probable R0 resection) than on oncologic criteria. The decision is simple for young patients but more difficult for older patients in whom a negative hormone receptor status appears to be a contraindication. PMID- 12559449 TI - Prognostic significance of the coexpression of p53 protein and c-erbB2 in breast cancer. AB - BACKGROUND: The expressions of p53 protein and c-erbB2 have been thoroughly analyzed as prognostic factors for breast cancer. However, the prognostic significance of the coexpression of p53 protein and c-erbB2 remains controversial. METHODS: The immunohistochemical expression of p53 protein and c erbB2 was evaluated in 242 women with breast cancer. RESULTS: According to the combination of p53 protein and c-erbB2, a group negative for both (125 cases), a group positive for either one of these two parameters (99 cases) and a group positive for both (18 cases) were identified. The group positive for one factor had a significantly (P = 0.0045) worse disease-free survival (DFS) than the group negative for both, while the group positive for both had a significantly (P = 0.0023) worse DFS than the group positive for one factor. A multivariate analysis indicated that the relative risk of p53 protein alone and cerbB2 alone was 2.18 and 2.93, respectively, while the relative risk of the group positive for one factor and the group positive for both was 2.29 and 6.37, respectively . CONCLUSIONS: Breast cancer with a coexpression of p53 protein and c-erbB2 was thus found to have a more significant prognostic value than those with a positive expression for either of these two biological parameters, while the prognostic significance of these two parameters themselves remained independent of each other. PMID- 12559450 TI - The coming of age of our understanding of the enterohepatic circulation of bile salts. AB - Recent advances in molecular biology have greatly accelerated knowledge relating to the significance of the enterohepatic circulation of bile salts. This review highlights the role that both oxysterols and bile salts play as ligands which, when bound to nuclear hormone receptors, activate transcription factors that set into play feed-forward catabolism of cholesterol to bile salts and feedback control of bile acid synthesis. The nuclear hormone receptors, liver X receptor (LXR) and farnesoid X receptor (FXR) both combined as heterodimers with retinoid X receptor and with oxysterols and bile salts, respectively as their ligands, initiate powerful genetic controls over cholesterol and bile acid homeostatic mechanisms. LXR/RXR signals molecular control of feed-forward catabolism of cholesterol to bile acids while FXR/RXR initiates feedback control of bile acid synthesis. An additional nuclear hormone receptor, small heterodimer partner (SHP), is required to inhibit the competence factor, liver receptor homolog-1 to achieve repression of bile acid synthesis in the liver and in so doing SHP autoregulates its own function. Additionally, while bile acid synthesis is repressed, pool size is preserved by the action of FXR/RXR at both hepatic and intestinal levels, which genetically signals enhanced hepatocyte bile salt transport by the bile salt export pump (BSEP) and the ileal bile acid binding protein (IBABP) for ileal reabsorption. During activation of cholesterol catabolism, LXR/RXR enhances reverse cholesterol transport by increasing cholesterol efflux via the ABC-1 transporter from extrahepatic cells. This cholesterol is then taken up by high-density lipoprotein (HDL) and transported back to the liver for further cholesterol catabolism and elimination in bile. The genetic coordination of nuclear hormone receptor function within the territory of the enterohepatic of bile salts allows for normal cholesterol and bile acid homeostasis thereby preventing atherosclerosis. PMID- 12559451 TI - Splenocolic fistula in a patient with polycythemia vera. PMID- 12559452 TI - Distribution of hydatid cysts into the liver with reference to cystobiliary communications and cavity-related complications. AB - BACKGROUND: This study was designed to test a hypothesis that intrahepatic hydatid cyst location can effect the incidence of cystobiliary communications and the cavity-related complications. METHODS: A total of 121 cysts treated by conservative surgical methods in 113 patients were evaluated prospectively. Cysts were grouped as near to the liver hilum (segment I, III, IVb, V, and VI) and far from the hilum (segment II, IVa, VII, and VIII). RESULTS: There were 58 (48%) hilar and 63 (52%) peripheral cysts. We found more cystobiliary communications (48% versus 27%, P = 0.015), more biliary leakage (36% versus 10%, P <0.001), and more biliary fistula (12% versus 3%, P = 0.080) in the cysts near to the hilum than far from the hilum. Postoperative hospital stay was longer in the cysts near to the hilum (12.3 +/- 3.1 days) than the cysts far from the hilum (7.7 +/- 2.7 days, P = 0.022). CONCLUSIONS: The location of the hydatid cyst near to the liver hilum is a risk factor for the cystobiliary communications and the cavity related complications. PMID- 12559453 TI - Gas in the splanchnic circulation. PMID- 12559454 TI - Seasonal timing of sperm production in roe deer: interrelationship among changes in ejaculate parameters, morphology and function of testis and accessory glands. AB - Roe deer are seasonal breeders with a short rutting season from mid-July to mid August. The seasonality of reproductive activity in males is associated with cyclic changes between growth and involution of both testes and the accessory sex glands. This study characterizes morphological and functional parameters of these organs prior to, during and after breeding season in live adult roe deer bucks. Size and morphology of the reproductive tract was monitored monthly by transcutaneous (testes, epididymis) and transrectal (accessory glands) ultrasonography. Semen was collected by electroejaculation. Concentration, motility and morphological integrity of spermatozoa as well as the content of proteins and testosterone in semen plasma were evaluated. Proportions of haploid, diploid and tetraploid cells were estimated by flow cytometry in testicular tissue biopsies. Serum testosterone was measured by enzyme immunoassay. Most parts of the male reproductive tract showed distinct circannual changes in size and texture. These changes were most pronounced in the testes, seminal vesicles, and prostate. All reproductive organs were highly developed during the rut only. The volume of ejaculates, total sperm number and percentages of motile and intact spermatozoa also showed a maximum during this period and corresponded with high proportions of haploid cells in the testis. The highest percentages of tetraploid cells were found in the prerutting period. The production of motile and intact spermatozoa correlated with both the protein content of semen plasma and the concentration of testosterone in semen plasma and blood serum. These results suggest the importance of combined actions of the testes and accessory sex glands and the crucial role of testosterone in facilitating the optimal timing of intensified semen production to ensure sufficient numbers of normal spermatozoa in seasonal breeders. PMID- 12559455 TI - Gonadotropin exposure, salt storage and storage duration affect penetration of domestic cat oocytes by homologous spermatozoa. AB - Salt-stored domestic cat oocytes are routinely used to study sperm function in domestic and nondomestic felids. Our objectives were to assess the effects of in vitro maturation (IVM), salt storage and storage duration on penetration of domestic cat oocytes by homologous spermatozoa. In Experiment 1, domestic cat spermatozoa were coincubated with fresh immature oocytes, salt-stored (2-3 weeks) immature oocytes, or salt-stored (2-3 weeks) IVM oocytes matured in Minimum Essential Medium containing 0.1IU FSH and 0.1IU LH/ml (IVM1) or 0.5IU FSH and 2.2IULH/ml (IVM2). In Experiment 2, all oocytes were matured (IVM2) and inseminated fresh or after salt storage for 2-3 weeks, 2-3 months or 9 months. In Experiment 1, penetration of the outer zona pellucida (OZP) was greater (P<0.05) in salt-stored IVM2 oocytes than in salt-stored immature oocytes, whereas penetration of salt-stored IVM1 oocytes was intermediate (P>0.05). In Experiment 2, penetration of the OZP and inner zona pellucida (IZP) was higher (P<0.05) in fresh IVM2 oocytes than in salt-stored oocytes, and a higher (P<0.05) proportion of oocytes had IZP sperm after 2-3 weeks of storage than after 2-3 months. Penetration of the perivitelline space was higher (P<0.05) in fresh IVM2 oocytes than in oocytes stored for 2-3 weeks or 2-3 months. These results suggest that oocyte penetration is improved by IVM, but is impaired by exposure to salt storage solution and prolonged storage duration. PMID- 12559456 TI - Presence of caprine arthritis-encephalitis virus (CAEV) proviral DNA in genital tract tissues of superovulated dairy goat does. AB - Transmission of caprine arthritis-encephalitis virus (CAEV) is not completely understood and the vertical route of infection from the goat to the embryo or to the fetus needs to be investigated. This route of infection involves the presence of CAEV in the genital tract tissues. Prior studies have detected CAEV-infected cells in genital secretions and in flushing media recovered during embryo collection from infected goats. To specify the origin of these cells, we conducted a double-nested polymerase chain reaction (PCR) test on embryo flushing media and on mammary gland, mammary lymph node, synovial membrane, pelvic lymph node, uterus and oviduct tissues from 25 CAEV-infected (blood PCR positive) embryo donor goats for the presence of CAEV proviral DNA. The presence of proviral DNA was found in 22 of 25 mammary gland samples, 14 of 25 uterus samples, and in 16 of 25 oviduct samples. Nineteen of 25 goats had at least one positive genital tract sample. Flushing media from 11 goats were PCR positive. All goats with positive-flushing media were oviduct positive. Of this group of does, except for 1 of the 11, infection of flushing media correlated with infection of almost all the other tissues examined. The frequency of positive tissues for flushing media-positive goats (61/66; 92%) was significantly higher than that for flushing media-negative goats (50/84; 60%) (P<0.01). This study demonstrated the presence of CAEV-infected cells in the goat genital tract. The presence of CAEV-infected cells in the uterus and oviducts suggests potential for vertical transmission of CAEV from doe to embryo or fetus. PMID- 12559457 TI - Efficacy of the anticaspase agent zVAD-fmk on post-thaw viability of canine spermatozoa. AB - Cryopreservation protocols for gametes are constantly improved with the aim of increasing the post-thaw viability of gametes. It is becoming clear that stress, resulting from cryopreservation, reduces cell numbers by apoptosis. Apoptosis, or programmed cell death, is a gene-activated event that occurs as a normal consequence of development and as a result of cellular stress. Apoptosis is mediated by the family of cysteine-dependent asparate-specific proteases (caspases). The present study was designed to test the hypothesis that addition of an anticaspase (zVAD-fmk) that has inhibitory properties against caspases and apoptosis to semen extenders and to the thaw medium would increase post-thaw viability of canine spermatozoa. Extenders were added in a two-step process. A dose of 100 microM caspase inhibitor was used. Four groups (n=6 for each) were composed based on the presence or absence of the caspase inhibitor: Group I (control), no caspase inhibitor in the extender or the thaw medium; Group II, caspase inhibitor in the thaw medium; Group III, caspase inhibitor in Extender II; and Group IV, caspase inhibitor in both Extender II and the thaw medium. Post thaw motility, plasma membrane integrity, and acrosome status were investigated. The addition of caspase inhibitor to Extender II or to the thaw medium failed to improve the parameters that were studied. The results suggest that this caspase inhibitor may not be beneficial to the post-thaw motility of canine spermatozoa if used at this concentration. PMID- 12559458 TI - Endocrine events during the periestrous period and the subsequent estrous cycle in ewes after estrus synchronization. AB - The aim of the present study was to investigate the endocrinology of the periestrus period and that of the subsequent estrous cycle in ewes synchronized during the breeding season. Animals were treated for 14 days with either MAP intravaginal sponges or subcutaneous progesterone implants, followed by administration of 500 IU PMSG at the time of withdrawal. The time to estrus occurrence following progestagen withdrawal differed significantly between groups (45.3+/-2.7h for the MAP and 21.5+/-1.2h for the implant group, P<0.001). Estradiol levels around estrus did not differ between groups, but a significant difference was detected for the interval from peak estradiol to estrus, with a shorter interval for the implant group (26.7+/-0.7 and 2.7+/-0.9h, P<0.001). Progesterone implants shortened the interval from removal to LH surge, compared to the MAP group (31.2+/-4.4 and 56.5+/-3.6h, respectively, P<0.05). An earlier response was also observed for the interval from estradiol peak to LH peak in the implant group (12.1+/-3.3 and 37+/-2h, respectively, P<0.005), but no difference was observed for the interval from estrus to LH surge. Progesterone levels, particularly during the Days 6 to 10 of the subsequent estrous cycle were significantly higher (P<0.05) in the implant group. It is concluded that the kind of progesterone treatment may affect the time of estrus and the LH peak as well as the progesterone levels of the subsequent cycle. PMID- 12559459 TI - Effects of methanol and developmental arrest on chilling injury in zebrafish (Danio rerio) embryos. AB - Stage-dependent chilling sensitivity has been reported for many species of fish embryos. Most of these studies reveal that developmental stages beyond 50% epiboly are less sensitive to chilling, but the chilling sensitivity accelerates rapidly at subzero temperatures. In this study, the effects of methanol and developmental arrest on chilling injury were studied using zebrafish (Danio rerio) embryos at 64-cell, 50% epiboly, 6-somite, prim-6 and long-bud stages. Embryos were exposed to methanol or anoxic conditions before they were cooled to 0 or -5 degrees C with slow (1 degrees C/min), medium (30 degrees C/min) or fast ( approximately 300 degrees C/min) cooling rates and were held at these temperatures for different time periods. Embryo survival was evaluated in terms of the percentage of treated embryos with normal developmental appearance after 3 day culture. Experiments on the effect of methanol on chilling sensitivity of the embryos showed that the addition of methanol to embryo medium increased embryo survival significantly at all developmental stages and under all cooling conditions. Higher concentration of methanol treatment generally improved embryo survival when embryos were cooled at a fast cooling rate of 300 degrees C/min. Experiments on the effect of developmental arrest on chilling sensitivity of embryos showed that embryos at 50% epiboly and prim-6 stages underwent developmental arrest almost immediately after 15 min oxygen deprivation. After 4h in anoxia, the survival rates of the embryos were not significantly different from their respective aerobic controls. Anoxia and developmental arrest had no effect on the chilling sensitivity of zebrafish embryos. PMID- 12559460 TI - Normal telomere lengths of spermatozoa in somatic cell-cloned bulls. AB - Interesting questions have been raised regarding cloned animals, including whether cloning restores cellular senescence undergone by donor cells, and how long cloned animals will be able to live. In this study, focusing our attention on the fact that telomere lengths of spermatozoa are longer than those of any somatic cells and that telomere length is maintained throughout aging in humans, we compared the telomere lengths of spermatozoa in normal and two somatic cell cloned cattle. The telomere lengths of the spermatozoa in the normal cattle (22.42+/-0.32 kb) were maintained throughout aging as in humans. In the cloned cattle, telomere lengths of the spermatozoa (25.8 and 20.9 kb) were the same as or longer than those found in normal cattle. Considering that telomere lengths of the donor cells, which had been derived from the muscle tissue of an old bull, were reported to be 20.1 kb, the results suggested that the telomere lengths of the germ cell line had extended from nucleus transfer to spermatogenesis. Moreover, we produced offspring (nine calves) from a somatic cell-cloned bull and measured the telomere lengths of their leukocytes. In all of the offspring, the telomere lengths of leukocytes were normal, too. These results indicate the possibility that somatic cloned bulls could be used as breeding sires. PMID- 12559461 TI - Timing of nuclear maturation of nonstored and stored domestic cat oocytes. AB - In this study we compared the effects of preculture storage of ovaries, IVM medium, a reduced O(2) atmosphere and duration of culture on in vitro maturation (IVM) of domestic cat oocytes. One randomly selected ovary of each pair (69 pairs) was stored in PBS at 10 degrees C for 16-24h before oocyte recovery. The second ovary from each pair was used as a nonstored control. In Experiment I, we investigated the effect of culture media (TCM 199 versus SOF) and a reduced O(2) atmosphere (a humidified gas atmosphere of either 5% CO(2) in air or 5% CO(2):5% O(2):90% N(2)) on IVM of both stored and nonstored oocytes. In the second experiment, we compared timing of nuclear maturation of both stored and nonstored oocytes cultured for 17-18, 20-21, 24-26, 28-30, 33-34 or 42-45 h before being evaluated for meiotic status. In both, Experiments I and II, the recovery rate, quality and competence for maturation of oocytes originating from stored ovaries did not differ (P>0.05) compared with nonstored. In Experiment I, neither culture medium (37.5 versus 43.2% of Metaphase II, respectively in TCM 199 versus SOF) or gas atmosphere (40.0 versus 32.5% of Metaphase II, respectively in 5% CO(2) in air versus 5% CO(2):5% O(2):90% N(2)) affected oocyte maturation. In Experiment II, the mean proportion of oocytes achieving Metaphase II within 17-18 h of culture was 36.1% and did not significantly increase (P>0.05) over time up to 28 h. The highest proportion of oocytes (67.3%) reached Metaphase II stage after 42 45 h of culture. Therefore, we conclude that two "waves" of nuclear maturation of cat oocytes can be distinguished. The first wave takes place within 26 h and it is likely that most oocytes of this wave mature by 17-18 h; the second wave occurs after 28-30 h of IVM. It can be assumed that this double wave may reflect the presence of two oocyte populations with two different degrees of "prematuration" which require different lengths of IVM. PMID- 12559462 TI - Effect of reducing sperm concentration during IVF on the ability to distinguish between bulls of high and low field fertility: work in progress. AB - The objectives of this study were to evaluate the effect of sperm dose and sire on the fertilization rate, cleavage rate and blastocyst yield following insemination in vitro, to examine the relationship between these parameters and field fertility in cattle, and to examine the relationship between blastocyst quality and sire used in IVF. Frozen semen from four bulls with 150-day nonreturn rates ranging from 57 to 78% was used. In Experiment 1, oocytes were inseminated with sperm from one of the four bulls at concentrations ranging from 0.016 to 0.5 x 10(6)sperm/ml. A proportion of presumptive zygotes were fixed at 17 h post insemination (hpi), while the remainder was transferred to in vitro culture (IVC) in droplets of synthetic oviduct fluid (SOF). Cleavage at 48 hpi and the percentage of oocytes reaching the blastocyst stage by Day 8 were recorded. In Experiment 2, to assess blastocyst quality, after insemination with semen from one of the four bulls, presumptive zygotes were cultured in SOF until Day 7. Blastocysts for each bull were removed and vitrified/warmed and survival was recorded at 24, 48 and 72 h after warming. Regardless of bull used, a concentration of 0.125 x 10(6)sperm/ml or above resulted in higher blastocyst yields than any lower concentration used. Fertilization and cleavage rates were also higher at higher sperm concentrations. The best predictor of field fertility was fertilization rate at a concentration of 0.5 x 10(6)sperm/ml (r=0.94, P<0.0001). There was also a significant correlation between cleavage rate at a concentration of 0.5 x 10(6)sperm/ml and nonreturn rate (r=0.90, P<0.0001). In Experiment 2, blastocysts derived from one bull, HTA, were of superior quality as measured by survival 24h after thawing, although these differences were less significant at the subsequent time points measured. In conclusion, these data show that differences between the field fertility of bulls can be determined at sperm concentrations routinely used in IVF. Lowering the sperm concentration does not increase the likelihood of optimizing the differences in fertility or cleavage rate between bulls of different field fertility. We have also demonstrated that the bull can have a significant effect on the quality of blastocysts produced using IVF techniques. PMID- 12559464 TI - A sporadic case of the sex-reversed mare (64,XY; SRY-negative): molecular and cytogenetic studies of the Y chromosome. AB - A sex-reversal syndrome appears frequently in the horse. The mare carriers of this syndrome lack of SRY gene. It is suggested that sex-reversal syndrome is probably caused by transfer of the SRY gene from Y to the X chromosome, due to abnormal meiotic exchange. The aim of the study was molecular analysis of the Y linked genes in a case of the sex-reversed infertile mare with 64,XY karyotype. The karyotype was established on the basis of analysis of 350 metaphase spreads stained by CBG banding. Molecular analysis of the loci assigned to the Y chromosome revealed absence of the SRY gene and presence of the other studied loci (ZFY, AMEL-Y and STS-Y). In this animal all fragments representing X chromosome (ZFX, AMEL-X and STS-X) were detected. External genitalia in the mare were normal, uterus was small and ovaries (examined by ultrasonography) extremely small. The mechanism of sex-reversal syndrome formation was discussed. It is postulated that during spermatogenesis in the sire two crossing-over events between the X and Y chromosomes occurred. One of them took place between the ZFY and SRY loci and another one between the SRY locus and the centromere. PMID- 12559465 TI - Birth of piglets after deep intrauterine insemination with flow cytometrically sorted boar spermatozoa. AB - The present study was carried out to determine the pregnancy rates, farrowing rates and litter size in sows with either induced or spontaneous ovulation inseminated with flow cytometric sorted spermatozoa using deep intrauterine insemination technology. Spermatozoa were stained with Hoechst 33342 and sorted by flow cytometry/cell sorting but not separated into separate X and Y populations. In Experiment 1, sows (n=200) were weaned and treated for estrus/ovulation induction with eCG/hCG. Inseminations with either sorted (70 or 140 million) or non-sorted (70 or 140 million) spermatozoa were done using a specially designed flexible catheter. Farrowing rates were 39.1 and 78.7% for 70 million of sorted and non-sorted, respectively, and 46.6 and 85.7% for 140 million of sorted and non-sorted, respectively (P<0.05). The litter size in sows inseminated with sorted spermatozoa showed a tendency to be lower than when non sorted spermatozoa were inseminated. In Experiment 2, sows (n=140) were inseminated as in Experiment 1 except that natural estrus was used. The ovaries of these sows were evaluated by transrectal ultrasonography. Farrowing rates were 25 and 77.2% for 70 million of sorted and non-sorted, respectively, and 32 and 80.9% for 140 million of sorted and non-sorted, respectively (P<0.05). These results show that the Deep Intrauterine Insemination technology can be successfully used to produce piglets from sorted spermatozoa when sows are hormonally treated to induce synchronous post weaning oestrus and ovulation. PMID- 12559463 TI - Influence of oxygen tension on apoptosis and hatching in bovine embryos cultured in vitro. AB - Various oxygen tensions are employed for in vitro embryo production. Since it is known that oxygen tension can influence the efficiency of embryo production and embryo quality, the aim of our study was to define an optimal oxygen concentration for bovine embryo production in vitro in synthetic oviduct fluid (SOF). Embryo quality criteria were hatching ability and the degree of apoptosis as assessed by TUNEL staining and Bax gene expression. In Experiment 1, the effects of 2, 5 and 20% O(2) tensions on embryo development were compared. The highest rate of eight-cell embryos (47%) at 72 hpi was obtained under 20% O(2). However, it seemed that 2 and 5% O(2) were also suitable as assessed by embryo survival rates at 144 hpi (29 and 30% at morula stage), 168 hpi (21 and 19% at blastocyst stage) and 216 hpi (14 and 17% at hatched blastocyst stage). In Experiment 2, comparisons were made between effects of 5, 20% and alternating O(2) (20% O(2) to 72 hpi and then changed to 5% O(2) up to 216 hpi) on embryo development. Alternating the O(2) tension significantly reduced the number of hatching blastocysts to 7%. Staining with TUNEL revealed that apoptosis occurred in all tested hatched blastocysts, but a significantly lower apoptotic cell ratio was found in embryos cultured under 5% O(2) (P<0.05). Total cell number of embryos cultured under 5% and alternating oxygen was significantly higher than that of other groups (P<0.05). Bax gene expression was detected by means of RT PCR in only 2 of 66 hatched blastocysts. It can be concluded that 5% oxygen is optimal for bovine embryo culture in cell free media. Moreover, it is very likely that the apoptosis detected by TUNEL staining in this study is Bax-independent. PMID- 12559467 TI - Evaluation of fresh and frozen-thawed semen of individual ganders by assessment of spermatozoa motility and morphology. AB - Individual differences in gander Anser anser L. reaction to semen collection procedure, quality and quantity of fresh semen and its susceptibility to the freezing process are discussed. Semen was collected individually by dorso abdominal massage, from 1-year old White Koluda ganders (n = 12) every 2-3 days. Ganders' reactions to massage were observed during the entire reproductive cycle (from 11 February to 13 June, from every male 40 semen collections were performed). For individual evaluation and freezing purpose semen was collected 13 times from every male. In the fresh semen, the following parameters were evaluated: ejaculate volume, color, density, blood or fecal contamination, motility, concentration and morphology of spermatozoa. Motility and spermatozoa morphology were evaluated in the frozen-thawed semen. Semen diluted in 2:1 ratio with EK diluent was frozen with 6% of dimethyl-formamide (DMF) to -140 degrees C at a rate 60 degrees C/min. Semen was thawed by placing the straws in a 60 degrees C water-bath for 4-5 s. Ten out of 12 ganders had from 67.5 to 100.0% positive reactions resulting in semen ejaculation. Significant (P < or = 0.01) differences in fresh semen quality of particular ganders were observed for all evaluated traits. In 1-year-old gander semen morphologically intact spermatozoa constitute only 27.8-45.2% of all cells. Therefore, the sperm quality factor (SQF), proposed by the authors, which includes ejaculate volume, sperm concentration and the percentage of live normal spermatozoa, seems to be a good predictor of gander semen fertilizing ability. The SQF of individual ganders varied from 7.7 to 11.5. The percentage of live normal spermatozoa in the frozen thawed semen depended mainly on fresh semen quality. In relation to the fresh semen average from 57.2 to 63.2% of spermatozoa survived freezing process and from 23.9 to 38.5% remained morphologically intact. PMID- 12559466 TI - In vitro and in vivo comparison of Ham's F-10, Emcare holding solution and ViGro holding plus for the cooled storage of equine embryos. AB - Equine embryos have been successfully transferred after 24h cooled storage in Ham's F-10. The aim of this study was to compare the viability of equine embryos in vitro and in vivo after 6 and 24h cooled storage using three media and to examine the relationship between embryo size and viability after 24h cooled storage. In Experiment 1, the viability of embryos was evaluated using DAPI staining after 0, 6 or 24h in Ham's F-10, 24h in Emcare embryo holding solution (EHS) or 24h in ViGro holding plus (VHP) (n=10/group). The mean number of dead cells was similar for embryos stored in Ham's F-10, EHS and VHP for 24h. Larger Day 7 embryos appear to withstand 24h cold storage better than small Day 7 embryos. The embryo quality for 24h cold storage was negatively correlated with size. In Experiment 2, 40 embryos were stored (n=20/group) either in Ham's F-10 or in EHS then transferred as pairs in recipient mares. Fifteen of the 20 recipient mares (75%) were pregnant. Out of 17 surviving embryos, 9 embryos (53%) were stored in Ham's F-10 and 8 (47%) in EHS. These results suggest that EHS and VHP offer a good alternative to Ham's F-10 for 24h cooled storage of equine embryos and that larger embryos may have a better viability after 24h of cooled storage than smaller embryos. PMID- 12559468 TI - Effect of maturation media and oocytes derived from sows or gilts on the development of cloned pig embryos. AB - In order to develop a culture system and recipient cytoplasm that could improve the developmental competence of somatic cell nuclear transfer (SCNT) embryos for successful cloning of pigs, we evaluated the effect of donor oocytes and in vitro maturation (IVM) media on maturation of oocytes and developmental competence of SCNT embryos. In Experiment 1, oocytes derived from sows or gilts were matured in two IVM media (TCM-199 versus NCSU-23) and maturation of oocytes was evaluated by the status of chromatin configuration, the diameter of matured oocytes, the thickness of the zona pellucida, and the size of the perivitelline space (PVS). Sow oocytes matured in TCM-199 (S-TCM group) and NCSU-23 (S-NCSU group) showed significantly higher (P<0.05) maturation rates (S-TCM and S-NSCU, 86+/-4 and 82+/ 4%, respectively) when evaluated by metaphase-II status than the gilt oocytes matured in TCM-199 (G-TCM group, 71+/-3%) and in NCSU-23 (G-NCSU-23 group, 71+/ 3%). Oocyte diameter, the thickness of the zona pellucida, and the perivitelline space of sow oocytes (S-TCM and S-NCSU) were larger than those of gilt oocytes (G TCM and G-NCSU) after IVM (P<0.05). In Experiment 2, SCNT was performed, using in vitro-matured oocytes from each group as recipient cytoplasm and porcine fetal fibroblasts as karyoplasts. The reconstructed embryos were electrically fused and activated, and cleavage and blastocyst formation were monitored under a stereomicroscope. The total cell number of flattened blastocysts stained with 5 microM bisbenzimide on day 7 were counted. In addition, in vitro matured non enucleated oocytes were also electrically activated (parthenogenetic activation) and pronuclear formation was monitored. No difference in pronuclear formation rate after parthenogenetic activation and fusion rate after SCNT was observed among experimental groups. A significantly higher cleavage rate (P<0.05) was observed in S-TCM (69+/-4%) when compared with only G-NCSU (58+/-4%), but not with G-TCM (60+/-4%) or S-NCSU (68+/-4%). The rate of blastocyst formation was significantly higher (P<0.05) in sow oocytes (24% in S-TCM and S-NCSU), when compared to that observed in G-TCM (15%), and G-NCSU (14%). When the same source of oocytes was used, there was no significant difference in rate of blastocyst formation in the two culture media. Total cell number of blastocysts were not significantly different among experimental groups. In conclusion, the present study clearly demonstrated that sow oocytes have a greater developmental competence than gilt oocytes, regardless of the maturation medium examined. PMID- 12559469 TI - Lack of association of Mycobacterium avium subsp. paratuberculosis with oocytes and embryos from moderate shedders of the pathogen. AB - Paratuberculosis is a chronic and progressive disease of the intestine in ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map). The bacterium is transmitted to young animals, becomes manifest in adulthood and leads to economic losses. The aim of this study is to investigate if cows shedding Map possess oocytes and embryos that are carriers of the bacterium. New genetical material can enter the dairy farm using embryo transfer but the question as to whether this technique is safe with respect to transmission of paratuberculosis has yet to be addressed. We selected and bought 16 cows, all proven to be moderate shedders of the bacterium in the faeces immediately prior to the experiment but none were clinically sick. One sample of uterine content was collected from each animal by flushing the uterus on the day of heat and five samples of homogenised uterine tissue were collected on the eighth day of the same cycle by biopsy. In addition, 217 cumulus-oocyte complexes (COCs), ranging from 3 to 35 COCs per animal, were collected using ultrasound guided transvaginal puncture of the ovarian follicles (OPU). On the seventh day of the subsequent cycle 31 embryos were obtained using the classic technique of super ovulation induction, artificial insemination (AI), followed by flushing of the uterus. These embryos have been washed and trypsinised. Fourteen of the 16 cows were treated again for super ovulation in the subsequent cycle and 19 foetuses were collected by opening of the uterus after euthanasia on Days 35-49 of the cycle. All samples were cultured for presence of Map and checked every 2 months during 1 year for bacterial growth. None of the samples showed growth of Map after 12 months of culture. Pathological examination of the cows revealed different degrees of severity of pathological alterations of the intestinal tract and mesenteric lymph nodes. However, the results suggest that neither in vivo embryo's nor oocytes are carriers of the bacteria and do not form an extra risk at transfer. However, due to the limited size of the experiment (sample size of 16 cows), a certain margin for error remains. PMID- 12559471 TI - Supernatant protein factor and tocopherol-associated protein: an unexpected link between cholesterol synthesis and vitamin E (review). AB - Supernatant protein factor (SPF) is a recently cloned member of a family of cytosolic lipid-binding proteins that includes Sec14p, alpha-tocopherol transfer protein, and cellular retinal-binding protein. SPF stimulates the conversion of squalene to lanosterol in the downstream pathway for cholesterol biosynthesis, and overexpression of cloned SPF in hepatoma cells increases cholesterol synthesis. The mechanism of this stimulation has yet to be defined, but SPF appears to facilitate the transfer of squalene into and between intracellular membranes. The recent identification of SPF as alpha-tocopherol-associated protein (TAP) has called into question its long-standing association with cholesterol biosynthesis. TAP binds alpha-tocopherol, but not other isomers of tocopherol, with high affinity; in the presence of alpha-tocopherol TAP translocates to the nucleus and activates reporter gene transcription. Given the ability of alpha-tocopherol to down-regulate the expression of two scavenger lipoprotein receptors, SR-A and CD36, these observations raise some interesting questions regarding the role of SPF/TAP and vitamin E in cholesterol metabolism. PMID- 12559470 TI - Metabolic profiles and progesterone cycles in first lactation dairy cows. AB - This study investigated the ovarian function, metabolic profiles and fertility in first lactation Holstein-Friesian dairy cows (mean 305 day milk yield: 7417+/ 191kg, n=37). Reproductive profiles obtained from milk progesterone analysis were categorized into normal (n=17) and four abnormal profiles (delayed ovulation, DOV1, n=9; DOV2, n=2; persistent corpus luteum, PCL1, n=6; PCL2, n=4; 1: immediately post-calving, 2: subsequent cycles). Fifty-five percent of cows had abnormal profiles with half of these being categorized as DOV1. Fertility of DOV1 and DOV2 cows was reduced whereas PCL1 and PCL2 cows had similar reproductive competence to normal profile cows. DOV1 animals had higher milk energy values, lower energy balances, lower dry matter intakes (DMI) and greater body weight and body condition score (BCS) losses post-calving than normal profile animals. DOV1 animals also had lower insulin-like growth factor-I (IGF-I) and higher betahydroxybutyrate (BHB) concentrations and tended to have the lower insulin and glucose concentrations in the pre-service period than normal profile cows. All PCL animals had vulval discharges postpartum. Despite this, the DMI, body weight and BCS changes, IGF-I concentrations and fertility of PCL1 animals was similar to normal profile cows. In conclusion, the high prevalence of delayed ovulation post-calving (DOV1) in primiparous high yielding cows lasted long enough (71+/ 8.3 days) to have a detrimental impact on fertility and was associated with significant physiological changes. This study did not establish any detrimental effects of PCL profiles on fertility or production parameters. PMID- 12559472 TI - Increase of bile acids synthesis and excretion caused by taurine administration prevents the ovariectomy-induced increase in cholesterol concentrations in the serum low-density lipoprotein fraction of Wistar rats. AB - We examined the effect of dietary taurine on the concentrations of serum cholesterol and apolipoprotein in lipoprotein fractions of Six-month-old ovariectomized, which were used as a model of hypercholesterolemia in postmenopausal woman, or sham operated rats. Taurine significantly reduced the serum total and low-density lipoprotein cholesterol concentrations only in the ovariectomized rats. In contrast, taurine significantly lowered the serum apolipoprotein B concentration and serum very low-density lipoprotein apolipoprotein E concentration only in the sham operated rats. The serum total and high density lipoprotein-apolipoprotein E concentrations were significantly lower in the rats fed taurine than in those fed the control diet regardless of whether they had undergone ovariectomy. The esterified cholesterol level in the liver was significantly lower and the level of hepatic cholesterol 7 alpha hydroxylase activity was significantly higher in the rats fed taurine than in those fed the control diet. The total bile acids concentration in the feces and intestinal contents of rats fed taurine were significantly higher than those in rats fed the control diet regardless of whether they had undergone ovariectomy. In the sham-rats, taurine accelerated bile acid synthesis and excretion, thereby increasing cholesterol consumption. The increased cholesterol consumption might be compensated by accelerating cholesterol synthesis and/or reducing the synthesis and release of very low-density lipoprotein from the liver. But in the ovariectomized rats, although taurine also accelerated bile acid synthesis and excretion, cholesterol demand might be compensated by excess cholesterol in the blood. PMID- 12559473 TI - Effects of dietary fat and zinc on adiposity, serum leptin and adipose fatty acid composition in C57BL/6J mice. AB - Zinc (Zn) has been implicated in altered adipose metabolism, insulin resistance and obesity. The objective of this study was to investigate the effects dietary Zn deficiency and supplementation on adiposity, serum leptin and fatty acid composition of adipose triglycerides and phospholipid in C57BL/6J mice fed low fat (LF) or high-fat (HF) diets for a 16 week period. Weanling C57BL/6J mice were fed LF (16% kcal from soybean oil) or HF (39% kcal from lard and 16% kcal from soybean oil) diets containing 3, 30 or 150 mg Zn/kg diet (ZD = Zn-deficient, ZC = Zn control and ZS = Zn-supplemented, respectively). HF-fed mice had higher fat pad weights and lower adipose Zn concentrations than the LF-fed mice. The ZD and ZS groups had a reduced content of fatty acids in adipose triglycerides compared to the ZC group, suggesting that zinc status may influence fatty acid accumulation in adipose tissue. Serum leptin concentration was positively correlated with body weight and body fat, and negatively correlated with adipose Zn concentration. Dietary fat, but not dietary Zn, altered the fatty acid composition of adipose tissue phospholipid and triglyceride despite differences in Zn status assessed by femur Zn concentrations. The fatty acid profile of adipose triglycerides generally reflected the diets. HF-fed mice had a higher percentage of C20:4 n-6, elevated ratio of n-6/n-3, lower ratio of PUFA/SAT and reduced percentage of total n-3 fatty acids in adipose phospholipid, a fatty acid profile associated with obesity-induced risks for insulin resistance and impaired glucose transport. In summary, the reduced adipose Zn concentrations in HF-fed mice and the negative correlation between serum leptin and adipose Zn concentrations support an interrelationship among obesity, leptin and Zn metabolism. PMID- 12559474 TI - Oxidized lipid depresses canine growth, immune function, and bone formation. AB - Dietary oxidized lipids can increase oxidative stress and potentially contribute to a variety of disease syndromes. This research describes the first use of a canine model to assess the effects of dietary oxidized lipids on growth, antioxidant status, and some immune functions. Three groups of eight, two-month old coon-hound puppies were pair fed diets for 16 weeks. The control diet contained <50 ppm aldehydes, and two additional diets contained thermally oxidized lipids targeted to contain 100 ppm aldehydes (medium-oxidation) and 500 ppm aldehydes (high-oxidation). Dogs fed the high-oxidation diet weighed less than those from the medium-oxidation (P < 0.05) and control groups (P < 0.001) at the end of the study. Oxidized lipids reduced serum vitamin E levels, total body fat content, and bone appositional rate. At different time points of the study, peripheral blood neutrophils and monocytes from dogs fed the HO diet had reduced oxidative burst capacity and produced less superoxide and hydrogen peroxide when stimulated with phorbol esters compared to the control group. Lymphocyte blastogenesis in response to concanavalin A was suppressed by dietary oxidized lipid. This study indicates that dietary oxidized lipids negatively affect the growth, antioxidant status, and some immune functions of dogs. Importantly, some effects are evident at 100 ppm aldehydes in the diet, which is a moderate level of oxidation. The rapid growth and weight gain of the dog during the first 6 months of life may also provide a better model for assessing the risks of dietary oxidized lipid in children and adolescents than previously used rodent models. PMID- 12559475 TI - Effects of dietary carbohydrate on the development of obesity in heterozygous Zucker rats. AB - Rats carrying one copy of the fa allele are predisposed to diet-induced metabolic disturbances which contribute to hyperinsulinemia, obesity and dyslipidemia. To investigate the role of dietary carbohydrate and fat in the development of these conditions, we fed 6-week old male heterozygous (fa/+) lean rats carbohydrate free diets containing primarily saturated fat either ad libitum or pair-fed. These diets were compared to standard chow and to a high saturated fat mixed diet containing 10% energy from sucrose for 4 weeks. The carbohydrate-free diet resulted in significantly lower circulating glucose levels compared to all other groups (p = 0.006). Weight gain was negligible in the carbohydrate free groups compared to standard diet and 10% sucrose diet (p = 0.03). This was reflected in energy efficiency which was markedly reduced (90%) in the carbohydrate-free groups compared to the other groups (p = 0.04). Corresponding changes were noted in fat pad mass. The subscapular and epididymal fat pads were increased 42% and 44%, respectively, in animals consuming the 10% sucrose diet compared to all other groups (p < 0.01). Comparable changes in fatty acid synthase (FAS) mRNA were observed in response to the carbohydrate-free diet, which resulted in a 53% decrease in adipocyte FAS mRNA (p < 0.001). Addition of 10% sucrose to the diet completely reversed this effect resulting in a 69% increase in adipocyte FAS mRNA compared to the carbohydrate-free groups (p = 0.01). Similarly, hepatic FAS mRNA was elevated by 51% and 66% in the 10% sucrose and standard diet groups respectively, compared to the carbohydrate-free groups. Therefore, diets that contain minimal carbohydrate may minimize net lipid storage and adiposity. PMID- 12559477 TI - Fructose and related phosphate derivatives impose DNA damage and apoptosis in L5178Y mouse lymphoma cells. AB - Glycation between reducing sugars and amino groups of long-lived macromolecules results in an array of chemical modifications that may account for several physiological complications. The consequences of the reaction are directly related to the reactivity of the sugars involved, whether aldoses or ketoses, phosphorylated or non-phosphorylated. So far, most studies have been focused on glucose, while fructose, a faster glycating agent, attracted minor attention. We have recently demonstrated that under in vitro conditions fructose and its phosphate derivatives can modify plasmid DNA faster than glucose and its phosphate metabolites. In the present study we provide further evidences suggesting that fructose and its phosphate metabolites, at the tested conditions, are cytotoxic and inflict deleterious DNA modifications to L5178Y cells in culture. Damage was verified by viable cell counts, MTT assay, colony forming ability, induction of mutation in the thymidine kinase gene, internucleosomal DNA cleavage, and single strand breaks. The intensity of the tested sugars to impose damage increased significantly in the following order: sucrose = glucose 1 phosphate < glucose < glucose 6-phosphate < fructose 1-phosphate = fructose < fructose 6-phosphate. Aminoguanidine, an inhibitor of the glycation reaction, inhibited internucleosomal DNA cleavage. Taken together, these results suggest that fructose triggers deleterious modification in cultured cells through the glycation process, and thus should deserve more attention as an agent that may induce physiological complications. PMID- 12559476 TI - Effects of dietary soybean lecithin on plasma lipid transport and hepatic cholesterol metabolism in rats. AB - Dietary lecithin can stimulate bile formation and biliary lipid secretion, particularly cholesterol output in bile. Studies also suggested that the lecithin rich diet might modify hepatic cholesterol homeostasis and lipoprotein metabolism. Therefore, we examined hepatic activities of 3-hydroxy-3 methylglutaryl coenzyme A reductase "HMG -CoA reductase", cholesterol 7 alpha hydroxylase and acyl-CoA: cholesterol acyltransferase "ACAT" as well as plasma lipids and lipoprotein composition in rats fed diets enriched with 20% of soybean lecithin during 14 days. We also evaluated the content of hepatic canalicular membrane proteins involved in lipid transport to the bile (all P-glycoproteins as detected by the C 219 antibody and the sister of P-glycoprotein "spgp" or bile acid export pump) by Western blotting. As predicted, lecithin diet modified hepatic cholesterol homeostasis. The activity of hepatic HMG-CoA reductase and cholesterol 7 alpha-hydroxylase was enhanced by 30 and 12% respectively, while microsomal ACAT activity showed a dramatic decrease of 75%. As previously reported from ACAT inhibition, the plasma level and size of very low-density lipoprotein (VLDL) were significantly decreased and bile acid pool size and biliary lipid output were significantly increased. The canalicular membrane content of lipid transporters was not significantly affected by dietary lecithin. The current data on inhibition of ACAT activity and related metabolic effects by lecithin mimic the previously reported effects following drug-induced inhibition of ACAT activity, suggesting potential beneficial effects of dietary lecithin supplementation in vascular disease. PMID- 12559478 TI - The association of preceding traumatic brain injury with mental disorders, alcoholism and criminality: the Northern Finland 1966 Birth Cohort Study. AB - The purpose of this study was to test the hypothesis that traumatic brain injury (TBI) during childhood and adolescence is associated with psychiatric disorders, heavy alcohol use and criminal offenses in adulthood. We made use of an unselected, general population birth cohort (n=12058) in Northern Finland, which was followed up prospectively up to the age of 31. The data on TBIs of the cohort members were collected from the hospital case notes of the outpatient clinics of the hospitals in the region and from the Finnish Hospital Discharge Registers (FHDR). The data on mental disorders including alcohol diagnoses were also collected from the FHDR after a careful validation process. The Ministry of Justice provided information on criminal offenses for all subjects. The final number of subjects in our study was 5589 males and 5345 females. We found that after controlling for confounders, TBI during childhood or adolescence increased the risk of developing mental disorders two-fold (OR 2.1, 95% CI 1.1-3.6) and TBI was significantly related to later mental disorder with coexisting criminality in male cohort members (OR 4.1, 95% CI 1.2-13.6). The results support the TBI's association with psychiatric morbidity, which should not be overlooked when treating psychiatric patients, especially those with comorbid criminality. PMID- 12559479 TI - Frontal Systems Behavior Scale in schizophrenia: relationships with psychiatric symptomatology, cognition and adaptive function. AB - Schizophrenia patients often exhibit impairments in executive functioning on formal testing and exhibit behaviors consistent with executive/frontal impairment in daily life. The Frontal Systems Behavior Scale (FrSBe) assesses behaviors associated with frontal lobe damage including executive dysfunction, apathy and disinhibition. We examined the reliability and validity of the FrSBe in 131 schizophrenia outpatients. Subjects were rated on the FrSBe and received symptom, cognitive and functional assessments. Statistical tests were corrected for multiple comparisons. The FrSBe was found to have good internal consistency and test-retest reliability. All three dimensions of the FrSBe (i.e. executive dysfunction, apathy and disinhibition) were significantly correlated with poor adaptive functioning as measured by the Social and Occupational Functioning Scale and the Functional Needs Assessment. In addition, differential relationships were found for apathy and disinhibition with symptoms as rated from the Brief Psychiatric Rating Scale and with cognitive variables including Trails B and verbal fluency scores. A multivariate analysis of variance examined differences on the FrSBe between patients and a group of 51 education-matched controls. Patients had significantly greater impairment on the FrSBe than controls. These differences were found for all FrSBe subscales. Results support the use of the FrSBe to characterize goal-directed behavior in schizophrenia patients. PMID- 12559480 TI - Acute L-5-hydroxytryptophan administration inhibits carbon dioxide-induced panic in panic disorder patients. AB - Previous research showed that lowering the availability of serotonin to the brain by tryptophan depletion increases the vulnerability of panic disorder patients for an experimental 35% CO(2) panic challenge. The results also suggested that increased availability of serotonin inhibits the response to such a challenge. In the present study, this latter possibility is examined. The reaction of 24 panic disorder patients and 24 healthy volunteers to a 35% CO(2) panic challenge was assessed following administration of 200-mg L-5-hydroxytryptophan (the immediate precursor of serotonin) or placebo. L-5-Hydroxytryptophan significantly reduced the reaction to the panic challenge in panic disorder patients, regarding subjective anxiety, panic symptom score and number of panic attacks, as opposed to placebo. No such effect was observed in the healthy volunteers. L-5 Hydroxytryptophan acts to inhibit panic, which supports a modulatory role of serotonin in panic disorder. PMID- 12559481 TI - Lack of a therapeutic effect of a 2-week sub-threshold transcranial magnetic stimulation course for treatment-resistant depression. AB - Stimulation parameters seem to strongly influence the efficacy of repetitive transcranial magnetic stimulation (rTMS) in the management of treatment-resistant depressed patients. The most effective and safest parameters are yet to be defined. Moreover, systematic follow-up data available to document the duration of the therapeutic effects remain sparse. Twenty-one treatment-resistant depressed patients were randomized to either active rTMS (N=12) or to sham (N=9) treatment in a double-blind design. Patients were kept on their medications. Sub motor-threshold (MT) stimulation (80% MT) was delivered for 10 consecutive work days (20 Hz, 2-s trains, 20 trains). Subjects meeting pre-set criteria for responding were entered into a follow-up phase for up to 5 months. Utilizing the above stimulation parameters, we found no significant difference between groups. Six patients in the active group and one subject in the sham group met criteria for the follow-up phase. The period of time before subjects met criteria for relapse was highly variable ranging from 2 to 20 weeks. Sub-threshold rTMS stimulation for 2 weeks is not significantly superior to sham treatment for treatment-resistant depressed patients. The duration of the therapeutic effects of rTMS delivered to the left prefrontal cortex using the above-described parameters is highly variable. PMID- 12559483 TI - Non-verbal interaction involvement as an indicator of prognosis in remitted depressed subjects. AB - Fifty-one remitted depressed inpatients and their interviewers were observed during a conversation. We investigated whether non-verbal behavioral elements indicative of involvement displayed by the remitted patients and/or their interviewers were predictive of depressive symptoms 6 months later. Involvement behavior of the patients appeared to be related to future complaints; the lower the level of involvement displayed, the more unfavorable the outcome. We interpret these results with reference to concepts of social support. PMID- 12559482 TI - Sexual diergism of baseline plasma leptin and leptin suppression by arginine vasopressin in major depressives and matched controls. AB - Leptin inhibits appetite by activating several neuroendocrine systems, including the hypothalamo-pituitary-adrenal cortical (HPA) axis. In turn, chronically elevated glucocorticoids increase circulating leptin. HPA axis hyperactivity occurs in 30-50% of patients with major depression, but the few prior reports of leptin measurements in this illness have shown inconsistent results. We, therefore, measured plasma leptin in 12 female and 8 male unipolar major depressives and 12 female and 8 male individually matched normal controls administered low-dose physostigmine (PHYSO) and arginine vasopressin (AVP) to stimulate the HPA axis. The subjects underwent four test sessions 5-7 days apart: PHYSO (8 microg/kg IV); AVP (0-08 U/kg IM); PHYSO+AVP; and saline control. Serial blood samples were taken before and after pharmacologic challenge and analyzed for leptin, ACTH(1-39), cortisol and AVP. Estradiol and testosterone also were measured at each test session. PHYSO and AVP produced no side effects in approximately half the subjects and predominantly mild side effects in the other half, with no significant patient-control differences. Correlations between side effects (absent or present) after PHYSO or AVP and the corresponding leptin responses were non-significant in all groups. Baseline plasma leptin concentrations (mean+/-S.D.) were significantly higher in the female patients compared to the female controls (22.5+/-13.9 ng/ml vs. 12.3+/-9.7 ng/ml), whereas they were similar in the male patients and the male controls (3.9+/-1.4 ng/ml vs. 3.6+/-2.0 ng/ml). Leptin concentrations following PHYSO remained unchanged from baseline, indicating that the short-lived ACTH and cortisol increases produced by PHYSO did not affect leptin secretion. In contrast, AVP administration, while also increasing ACTH and cortisol, significantly suppressed leptin, more so in the women than in the men. Baseline leptin and the leptin decrease after AVP were moderately positively correlated with the Hamilton Depression Scale 'somatization' factor in the female patients (r=0.50) and more strongly correlated with the 'mood-depression' factor in the male patients (r=0.81). These findings indicate a sexual diergism (functional sex difference) in plasma leptin measures between major depressives and matched normal controls. PMID- 12559485 TI - Use of telephone screens improves efficiency of healthy subject recruitment. AB - Previous reports have indicated that a high percentage of those responding to advertisements for healthy controls for psychiatric research have personal or family histories of illness which would exclude them from such studies. We reviewed 1757 telephone screen interviews conducted over 14 years to determine: (1) the effectiveness of a screen for excluding unhealthy volunteers, (2) whether the reasons for exclusion changed over time, and (3) the final yield of healthy participants for psychiatric research after taking account of exclusions from telephone screens, no-shows and exclusions identified by direct interviews. Volunteers for psychiatric research, solicited by 43 newspaper advertisements in the Baltimore metropolitan area from 1989-2002, were initially screened by telephone and scheduled for in-person interviews if no exclusions were identified at the initial screen. More than half of the telephone respondents had major medical illnesses, substance abuse problems, depression, an Axis I disorder (not depression), a relative with a psychotic disorder or were otherwise deemed not eligible for face-to-face interview. Of the telephone respondents scheduled for direct interviews, 29% did not show up. However, the respondents who completed the direct interviews had a high likelihood of inclusion as non-ill controls for psychiatric research studies. Since Axis I and II interviews and family history take approximately 4-5 h per subject, the telephone screen is a cost-effective initial step in identifying healthy controls for psychiatric research. PMID- 12559484 TI - Interpersonal deficits meet cognitive biases: memory for facial expressions in depressed and anxious men and women. AB - Memory biases for negatively vs. positively valenced linguistic information in depression are well documented. However, no study so far has examined the relationship between depression and memory for facial expressions. We examined memory for neutral, happy, sad, and angry facial expressions in individuals suffering from comorbid depression and anxiety (COMs, N=23) or from anxiety disorders (ANXs, N=20) and in normal controls (NACs, N=23). Two main hypotheses were examined. First, we expected COMs, but not NACs, to exhibit an enhanced memory for sad and angry vs. happy expressions (negativity hypothesis). Second, we postulated that this bias would be specific to depression (disorder specificity hypothesis). Data supported both these hypotheses. Specifically, COMs exhibited enhanced recognition of angry compared to happy expressions; in contrast, ANXs and NACs did not exhibit such enhancement. We also found that men showed a significantly better memory for angry vs. sad expressions, whereas women did not exhibit such a difference. The implications of these findings for the interpersonal processes involved in the maintenance of depression and anxiety are discussed. PMID- 12559486 TI - Sentence completion test in combat veterans with and without PTSD: preliminary findings. AB - This study used a sentence completion task to assess semantic choice in combat veterans. Twenty-eight combat veterans with (n=14) and without (n=14) posttraumatic stress disorder (PTSD) filled in the final word for 33 incomplete sentences after receiving a combat prime. The veterans with PTSD completed sentences with significantly more trauma-relevant final words than those without PTSD. Findings are interpreted with respect to current language models and information-processing theories of PTSD. PMID- 12559487 TI - Equarin, a novel soluble molecule expressed with polarity at chick embryonic lens equator, is involved in eye formation. AB - The lens plays an important role in eye development. To investigate the molecular mechanisms involved, we used signal sequence trap screens with a chicken lens cDNA library and identified a novel secreted molecule, equarin. Equarin encodes consensus repeat domains conserved in human SRPX and mouse Urb. In the embryonic eye, equarin transcript is detected exclusively in the lens, and persists in the lens equatorial region in a high-dorsal-to-low-ventral gradient. In vitro analysis of equarin protein indicated that after translation, it is modified, cleaved, and secreted to extracellular locations. Microinjection of equarin mRNA into Xenopus embryos induced abnormal eye development. These data suggest that equarin is involved in eye formation. PMID- 12559488 TI - Armadillo levels are reduced during mitosis in Drosophila. AB - The Armadillo protein of Drosophila melanogaster is both a structural component of adherens junctions at apical cell membranes and also a key cytoplasmic transducer of the Wingless signalling pathway. We have used the Gal4-UAS system to over-express Armadillo in the Drosophila wing: this hyperactivates the Wingless pathway and leads to the formation of ectopic, supernumerary wing bristles. Here, we report that this adult phenotype is dominantly enhanced by mutations in cdc25(string) and, conversely, is suppressed by co-expression of Cdc25(String). Furthermore, we show that the steady state levels of Armadillo protein produced from the UAS transgene are also sensitive to cdc25(string) dosage in the cells of the larval imaginal wing disc. Consistent with the role of Cdc25(String) in promoting mitosis and with our genetic interaction data, we find a strong correlation between progression through mitosis and a reduction in Armadillo levels. Significantly, this is true whether Armadillo is over-expressed or not, and both cytoplasmic (signalling) and membrane-associated (junctional) Armadillo appears to be affected. We conclude that this phenomenon may reduce the efficacy of Wingless signalling and/or intercellular adhesion during cell division. PMID- 12559489 TI - The IIIb isoform of fibroblast growth factor receptor 2 is required for proper growth and branching of pancreatic ductal epithelium but not for differentiation of exocrine or endocrine cells. AB - Fibroblast growth factors (Fgfs) and their receptors have been implicated in embryonic pancreas development. Recently it was shown that Fgf10, a major ligand for the IIIb isoform of fibroblast growth factor receptor 2 (Fgfr2b), has an important regulatory role in early pancreas development. The aim of our study was to define the role of Fgfr2b in pancreas development by analyzing the phenotype of Fgfr2b (-/-) mice. Pancreases of Fgfr2b (-/-) embryos were noticeably smaller than the wild type littermates during embryogenesis, and pancreatic ductal branching as well as duct cell proliferation was significantly reduced. However, both exocrine and endocrine pancreatic differentiation occurred relatively normally. Exogenous addition of Fgfr2b ligands (Fgf7 and Fgf10) stimulated duct cell proliferation and inhibited endocrine cell differentiation in the ex vivo embryonic organ cultures of wild type pancreas. Our results thus suggest that Fgfr2b-mediated signaling plays a major role in pancreatic ductal proliferation and branching morphogenesis, but has little effect on endocrine and exocrine differentiation. PMID- 12559490 TI - Pax6 in the sepiolid squid Euprymna scolopes: evidence for a role in eye, sensory organ and brain development. AB - The cloning of a Pax6 orthologue from the sepiolid squid Euprymna scolopes and its developmental expression pattern are described. The data are consistent with the presence of a single gene encoding a protein with highly conserved DNA binding paired and homeodomains. A detailed expression analysis by in situ hybridization and immunodetection revealed Pax6 mRNA and protein with predominantly nuclear localization in the developing eye, olfactory organ, brain lobes (optic lobe, olfactory lobe, peduncle lobe, superior frontal lobe and dorsal basal lobe), arms and mantle, suggestive of a role in eye, brain, and sensory organ development. PMID- 12559491 TI - Wolbachia distribution and cytoplasmic incompatibility during sperm development: the cyst as the basic cellular unit of CI expression. AB - The growth and distribution of the intracellular microbe Wolbachia pipientis during spermatogenesis in several different host/symbiont genetic combinations in Drosophila melanogaster and Drosophila simulans is described. Considerable intra- and inter-strain variation in Wolbachia density and tissue distribution was observed. Wolbachia were found inside spermatocytes and spermatids or within the somatic cyst cells surrounding the germ cells. Some strains displayed both tissue distributions. High rates of cytoplasmic incompatibility (CI) are correlated with high levels of Wolbachia only when spermatocytes and/or spermatids harbor the microbe. Wolbachia infection of somatic cyst cells, although sometimes present at high levels, did not result in significant CI expression. CI-inducing Wolbachia strains within D. simulans showed no distinguishable differences in distribution or density within infected spermatids. To dissect the relative contribution of host and symbiont to the expression of CI, Wolbachia from various host strains known to exhibit varying levels of CI were introgressed into new uninfected host genetic backgrounds. These introgression experiments confirm that the mod(+)/mod( ) phenotype is an intrinsic Wolbachia trait and is not determined by host factors. The level of sperm modification in those lines harboring Wolbachia capable of modifying sperm, however, is influenced by host genetic background. These results form the basis of the Wolbachia Infected Spermatocyte/Spermatid Hypothesis (WISSH). According to WISSH, Wolbachia infection in spermatocytes and then spermatids during sperm development is required for CI expression. PMID- 12559492 TI - Expression of rigf, a member of avian VEGF family, correlates with vascular patterning in the developing chick limb bud. AB - In a differential display screening for genes regulated by retinoic acid in the developing chick limb bud, we have isolated a novel gene, termed rigf, retinoic acid induced growth factor, that encodes a protein belonging to the vascular endothelial growth factor (VEGF) family. Rigf transcripts were found in the posterior region of the limb bud in a region-specific manner as well as in other embryonic tissues and regions, including the notochord, head and trunk mesenchyme, retinal pigment epithelium, and branchial arches. Several manipulations revealed that retinoic acid and sonic hedgehog signaling pathways regulate rigf expression in the limb bud. VEGF family members, which promote the migration, differentiation and proliferation of endothelial cells in both blood and lymphatic vessels, are important factors for the formation of blood and lymphatic vasculatures during development. We demonstrated that the anterior border of the rigf expression domain in the limb bud corresponds with the position of the primary central artery (the subclavian artery in the forelimb), which is a main artery for supplying blood to the limb. These observations taken together with results from some experimental manipulations suggest that the limb tissue attracts blood vessels into the limb bud and that rigf is involved in the pattern formation of blood vessels in the limb. PMID- 12559493 TI - Multiple regulatory elements with spatially and temporally distinct activities control neurogenin1 expression in primary neurons of the zebrafish embryo. AB - The basic Helix-Loop-Helix gene neurogenin1 (ngn1) is expressed in a complex pattern in the neural plate of zebrafish embryos, demarcating the sites of primary neurogenesis. We have dissected the ngn1 locus to identify cis-regulatory regions that control this expression. We have isolated two upstream elements that drive expression in precursors of Rohon-Beard sensory neurons and hindbrain interneurons and in clusters of neuronal precursors in the anterior neural plate, respectively. A third regulatory region mediates later expression. Thus, regulatory sequences with temporally and spatially distinct activities control ngn1 expression in primary neurons of the zebrafish embryo. These regions are highly similar to 5' sequences in the mouse and human ngn1 gene, suggesting that amniote embryos, despite lacking primary neurons, utilize related mechanism to control ngn1 expression. PMID- 12559494 TI - Krapfen/dMyd88 is required for the establishment of dorsoventral pattern in the Drosophila embryo. AB - In Drosophila, the dorsoventral axis is set up by the action of the dorsal group of genes and cactus, which have been ordered genetically in a linear pathway. We have identified and characterised krapfen (kra) as a new member of the dorsal group genes. kra encodes for the Drosophila homologue of MyD88, an adapter protein operating in the mammalian IL-1 pathway. Epistasis experiments reveal that kra acts between the receptor Toll and the cytoplasmic factor Tube. We show that there is a direct interaction between Kra and Tube presumably mediated by the death domains present in both proteins. Tube in turn interacts with its downstream effector Pelle through death domain association. We therefore suggest that upon Toll activation, Kra associates with Pelle and Tube, in an heterotrimeric complex. PMID- 12559496 TI - The OAR/aristaless domain of the homeodomain protein Cart1 has an attenuating role in vivo. AB - Aristaless-related genes encode a structurally defined group of homeoproteins that share a C-terminal stretch of amino acids known as the OAR- or aristaless domain. Many aristaless-related genes have been linked to major developmental functions, but the function of the aristaless domain itself is poorly understood. Expression and functional studies have shown that a subgroup of these genes, including Prx1, Prx2, Alx3, Alx4 and Cart1, is essential for correct morphogenesis of the limbs and cranium. We now demonstrate the function of the aristaless domain in vivo by ectopically expressing normal and mutated forms of Cart1 and Alx3. Ectopic expression of Cart1 in transgenic mice does not disturb development, whereas expression of a Cart1 form from which the aristaless domain has been deleted results in severe cranial and vertebral malformations. The Alx3 protein contains a divergent aristaless domain that appears not to be functional, as ectopic expression of Alx3 results in an altered phenotype irrespective of the presence of this aristaless domain. Linking the Cart1 aristaless domain to Alx3 extinguishes teratogenicity. We show that, at the molecular level, the most important consequence of deleting the aristaless domain is increased DNA binding to its palindromic target sequence. This demonstrates that the aristaless domain functions as an intra-molecular switch to contain the activity of the transcription factor that it is part of. PMID- 12559495 TI - The lateral somitic frontier: dorso-ventral aspects of anterio-posterior regionalization in avian embryos. AB - Patterning events along the anterior-posterior (AP) axis of vertebrate embryos result in the distribution of muscle and bone forming a highly effective functional system. A key aspect of regionalized AP patterning results from variation in the migratory pattern of somite cells along the dorsal-ventral (DV) axis of the body. This occurs as somite cell populations expand around the axis or migrate away from the dorsal midline and cross into the lateral plate. The fate of somitic cells has been intensely studied and many details have been reported about inductive signaling from other tissues that influence somite cell fate and behavior. We are interested in understanding the specific differences between somites in particular AP regions and how these differences contribute to the global pattern of the organism. Using orthotopic transplants of segmental plate between quail and chick embryos, we have mapped the interface of the somitic and lateral plate mesoderm during the formation of the body wall in cervical and thoracic regions. This interface does not change dramatically in the mid-cervical region, but undergoes extensive changes in the thoracic region. Based on this regional mapping and consistent with the extensive literature, we suggest a revised method of classifying regions of the body wall that relies on embryonic cell lineages rather than adult functional criteria. PMID- 12559497 TI - Mitotic G2-arrest is required for neural cell fate determination in Drosophila. AB - In the wing discs of Drosophila, the mecanosensory precursor cells are singled out from clusters of cells blocked at the G2 phase of the cell cycle. This mitotic quiescence and the selection of the precursors are under strict spatio temporal control. We forced G2 cells to enter mitosis by overexpression of string, the Drosophila homologue of the cdc25 gene. Premature entrance in the cell cycle is associated to a loss of precursor cells. Precursors are lost consecutively to a transcriptional down-regulation of the determinant proneural achaete/scute genes. This down-regulation results from an over-activation of the Enhancer of Split genes, known as effectors of the Notch signalling pathway. We conclude that exit from the cell cycle is required for proper neural cell fate determination. PMID- 12559498 TI - The role of intravenous immunoglobulin in the treatment of multiple sclerosis. AB - Intravenous immunoglobulin (IVIG) has several effects on the immune system that could have a beneficial influence on disease processes in multiple sclerosis (MS). Owing to its anti-inflammatory properties, IVIG may be beneficial in the treatment of acute relapses and in prevention of new relapses. By promoting remyelination, IVIG could have a beneficial effect on disability and disease progression. Four double-blind trials in relapsing-remitting MS have demonstrated that IVIG reduces the relapse rate and the number of gadolinium enhancing lesions, and in this respect seems comparable to established therapies in relapsing-remitting MS, i.e. interferon-beta and glatiramer acetate. The doses of IVIG that have been used for treatment in relapsing-remitting have varied 10 fold, and the ideal dosage of IVIG for treating MS still needs to be determined. Three studies have been performed to assess the effect of IVIG on chronic visual impairment or established motor symptoms in MS. None of these trials could confirm that established symptoms in MS can be reversed by IVIG. In secondary progressive MS, a large randomized placebo-controlled trial has recently shown that IVIG is without beneficial effects in this phase of the disease. In conclusion, IVIG is a valuable alternative for treatment of relapsing-remitting MS in patients who do not tolerate or are unwilling to take the approved injectable medications, but additional studies are needed to establish the role of IVIG in the management of multiple sclerosis. PMID- 12559499 TI - Clinical and laboratory characteristics of secondary progressive MS. AB - Secondary progressive (SP) MS follows on from but is distinct in its clinical picture from relapsing remitting (RR) MS. Diagnosis is usually straightforward except during the transitional stage when the two phenotypes merge. It is clear that most patients that start with relapsing remitting MS will develop SP disease, although the underlying pathogenesis that causes this change is subject to much debate. Clinical features such as pattern and site of symptoms, and age of onset, in the relapsing remitting stage versus progressive disease, suggests a difference in the pathophysiology. Laboratory markers may give insight into the disease mechanisms. Measures of urinary and CSF myelin basic protein-like material (MBPLM) indicate demyelination and subsequent oligodendrocyte and axonal loss. Tertiary neutralising antibodies to MBP antibodies could attenuate remission and lead to continuous progression, and neuronal antibodies found in SP disease may contribute to the axonal loss. In addition, differences in nitric oxide and other inflammatory cytokine patterns might either be secondary to or causative of the pathological mechanisms.Greater understanding of progressive MS is a priority considering permanent disability results almost entirely from this stage of the disease. PMID- 12559500 TI - Prognostic factors for progression of disability in the secondary progressive phase of multiple sclerosis. AB - Predicting clinical outcome is one of the major and most interesting issues in MS patients. If the global evolution of disability (usually chosen levels on the Kurtzke's Disability Status Scale) has been widely studied, much less is known about the progression of disability during the secondary progressive phase of the disease. It is usually said that there is a poorer prognosis once patients enter a progressive phase, whether from onset (like in primary progressive MS) or after an initial relapsing-remitting phase. The secondary progressive phase of multiple sclerosis (MS) is the one most often associated with the development of severe and irreversible disability. Despite this fact, there is a lot of information about the initial relapsing-remitting phase in the literature, but much less about the secondary progressive one. We review here information available from the literature and from the Lyon MS database about this secondary progressive phase. PMID- 12559501 TI - Clinical-MRI correlations in the secondary progressive phase of MS: lessons from the treatment trials. AB - Conventional MRI techniques are sensitive to detect MS lesions and their change overtime. In relapsing-remitting MS correlations with clinical measures are weak suggesting a pathological heterogeneity of these lesions. There are less data available in secondary progressive phase of the disease. The best source for clinical MRI correlations analysis is the placebo arm of the published interferon beta trials. This review presents the main clinical-MRI findings from these trials then focuses on recent promising observations obtained with non conventional MRI techniques in SP MS patients. PMID- 12559502 TI - The diagnosis of primary progressive multiple sclerosis. AB - Primary progressive multiple sclerosis (PPMS) is a rather unique form of the more common relapsing inflammatory demyelinative disease. The absence of attacks that typify relapsing forms of MS imposes special challenges for diagnosis, but also provides an opportunity to study the pathogenesis of the more progressive aspects of the disease process in isolation of confounding transient clinical events. In this review, recent advances in diagnostic approaches are considered in relationship to baseline data from a large multinational study designed to better characterize and treat this clinical phenotype. PPMS subjects with cerebral spinal fluid (CSF) findings consistent with intrathecal immunoglobulin production may have a more tissue destructive disease process than those whose CSF lacks evidence of a B-cell immunopathogenic disease component. PMID- 12559503 TI - Primary and secondary progressive multiple sclerosis. AB - The progressive phase of multiple sclerosis (MS) is the one most often associated with irreversible accumulation of disability. An important question remains about the place of primary progressive MS (PP-MS): does it form an integral part of the disease spectrum, or is it maybe a distinct entity? This question could apparently be very theoretical, but it is not, as patients with PP-MS remain orphans when regarding disease-modifying treatments. Thus, they are usually excluded from therapeutic trials. A clue to this question could be the comparison between the different MS subtypes with a progressive phase. We discuss here the clinical similarities and differences between secondary and primary progressive MS. PMID- 12559504 TI - MRI-clinical correlations in the primary progressive course of MS: new insights into the disease pathophysiology from the application of magnetization transfer, diffusion tensor, and functional MRI. AB - Despite patients with primary progressive multiple sclerosis (PPMS) experience a progressive disease course from onset, the burden and activity of lesions on conventional magnetic resonance imaging (MRI) scans of the brain are lower than in all other main clinical phenotypes of MS. This review outlines the major contributions given by magnetization transfer MRI, diffusion tensor MRI and functional MRI to the understanding of the pathophysiology of PPMS and provides evidence that, at least, three factors might explain this clinical/MRI discrepancy: (a) the presence of a diffuse tissue damage at a microscopic level; (b) a prevalent involvement of the cervical cord, and (c) an impairment of the adaptive capacity of the cortex to limit the functional consequences of subcortical structural damage. PMID- 12559505 TI - Axonal loss in the pathology of MS: consequences for understanding the progressive phase of the disease. AB - Axonal degeneration has been identified as the major determinant of irreversible neurological disability in patients with multiple sclerosis (MS). Axonal injury begins at disease onset and correlates with the degree of inflammation within lesions, indicating that inflammatory demyelination influences axon pathology during relapsing-remitting MS (RR-MS). This axonal loss remains clinically silent for many years, and irreversible neurological disability develops when a threshold of axonal loss is reached and compensatory CNS resources are exhausted. Experimental support for this view-the axonal hypothesis-is provided by data from various animal models with primary myelin or axonal pathology, and from pathological or magnetic resonance studies on MS patients. In mice with experimental autoimmune encephalomyelitis (EAE), 15-30% of spinal cord axons can be lost before permanent ambulatory impairment occurs. During secondary progressive MS (SP-MS), chronically demyelinated axons may degenerate due to lack of myelin-derived trophic support. In addition, we hypothesize that reduced trophic support from damaged targets or degeneration of efferent fibers may trigger preprogrammed neurodegenerative mechanisms. The concept of MS as an inflammatory neurodegenerative disease has important clinical implications regarding therapeutic approaches, monitoring of patients, and the development of neuroprotective treatment strategies. PMID- 12559506 TI - Differences between subgroups of MS: MRI findings and correlation with histopathology. PMID- 12559507 TI - MRI-clinical correlations: more than inflammation alone-what can MRI contribute to improve the understanding of pathological processes in MS? AB - Magnetic resonance imaging (MRI) is a sensitive technique in the detection and follow-up of multiple sclerosis (MS) lesions. However, the pathologic basis of these changes is poorly understood. It is becoming increasingly apparent that routine MRI techniques do not represent specific histopathologic changes but reflect a variety of pathological changes in tissue. This review will outline the findings in MRI-pathology correlation in MS to date, and the current understanding of the evolution of pathologic changes in the MS lesion and non lesional white matter as reflected by MRI. Possible future contributions of MRI to unveiling the dynamic pathology of MS will also be discussed. PMID- 12559508 TI - Remyelination in multiple sclerosis. AB - Remyelination in multiple sclerosis (MS) lesions has been described in several studies. It depends on the presence of myelinating oligodendrocytes and a functional interaction between these myelinating cells and axons. The imaging signal of remyelination in magnetic resonance imaging or spectroscopy is not yet defined. The present review will focus on the morphological appearance of remyelinating MS lesions, their correlation with oligodendrocyte pathology, and possible markers for remyelination in imaging. PMID- 12559509 TI - Hypoxia-like tissue injury as a component of multiple sclerosis lesions. AB - Recent data suggest that the mechanisms of demyelination and tissue damage in multiple sclerosis (MS) are heterogenous. In this review, evidence is discussed, which show that in a subset of multiple sclerosis patients the central nervous system (CNS) lesions show profound similarities to tissue alterations found in acute white matter stroke, thus suggesting that a hypoxia-like metabolic injury is a pathogenetic component in a subset of inflammatory brain lesions. Both, vascular pathology as well as metabolic disturbances induced by toxins of activated macrophages and microglia may be responsible for such lesions in multiple sclerosis. PMID- 12559510 TI - Neurophysiological evidence for long-term repair of MS lesions: implications for axon protection. AB - After recovery from the acute stage of optic neuritis, a marked prolongation in the latencies of visual evoked potentials (VEPs) is typically observed. We have conducted three studies (one cross-sectional, two prospective), aimed at elucidating the progressive shortening of VEP latency, which frequently ensues over the following months or years. This has been shown to be a progressive process and a prevalent tendency in the patient population, proceeding for more than 2 years in spite of the fact that very little functional improvement in vision occurs after the first few months. We argue that the underlying process of repair is most likely to involve remyelination of demyelinated optic nerve axons. Rather than restoration of visual function (which may be virtually complete after as short a period as 3 months), the main importance of the long-term myelin repair process may consist in protecting demyelinated axons from subsequent, permanent degeneration. In the VEPs of the acutely unaffected fellow eyes followed up over 3 years, we observed an asymptomatic deterioration, possibly due to insidious processes of demyelination and/or axonal degeneration. Even in the relapsing/remitting stage of MS, therefore, there is electrophysiological evidence for involvement of clinically asymptomatic axons, which, in the later stages, may be manifested as progressive functional deterioration. PMID- 12559512 TI - Mitoxantrone in progressive multiple sclerosis: when and how to treat? AB - Mitoxantrone (MX) has been approved by the Food and Drug Administration (FDA) for the treatment of patients with worsening relapsing-remitting (RR) or secondary progressive (SP) multiple sclerosis (MS). However, indications should be refined and mitoxantrone reserved as a rescue therapy to: (1) patients in the relapsing remitting phase with frequent and disabling exacerbations likely leading to permanent severe disability and (2) to patients in the secondary progressive phase whose disability progression rate increases by one EDSS point or more per year and who do not respond to other current therapies. An induction phase with the monthly intravenous administration of 12 mg/m(2) followed by a maintenance phase with 12 mg/m(2) every 3 months for 2 years seems the most effective and safe treatment regimen, not exceeding the maximum cumulative dose of 140 mg/m(2). Given the potent myelosuppressive activity of mitoxantrone, dosage should be carefully adapted to the body surface and hematological changes. Long-term toxicities (amenorrhoea and therapy-related leukemia) seem acceptable but a valid evaluation will need a longer follow-up in more patients. Cardiotoxicity, the major long-term toxicity, is clearly dose-dependent and is a strict treatment duration limiting factor. To reduce the risk of cardiac events, the drug should be administered by slow infusion (over 30 min). Analogs of mitoxantrone with a much lower cardiotoxicity are currently investigated in animal experimental models. PMID- 12559511 TI - Interferon beta 1a for secondary progressive multiple sclerosis. AB - This non-systematic review identified four randomised trials that have tested the efficacy of interferon beta in secondary progressive multiple sclerosis (SPMS). Two were trials of interferon beta 1a (IFNb1a) and two of interferon beta 1b (IFNb1b). All have shown significant reductions in relapse rates and accumulation of new magnetic resonance imaging (MRI) lesions, but only one trial (of IFNb1b) showed significant slowing of disability progression. Post hoc analyses of these trials suggest that the differences in outcomes might be partly explained by the differences between the trials in the proportions of patients with relapsing disease. In one of the trials of IFNb1a (the SPECTRIMS trial), the hazard ratio for progression in the treated relapsing patients with relapses in the two pre study years was 0.74 compared to placebo patients with pre-study relapses and 1.01 in the treated patients compared to the placebo patients without pre-study relapses. In the same trial, the treatment effects on MRI parameters were more marked in the patients who had recent pre-study relapses compared with those who had not. These observations have led to the recommendation in national guidelines that prescribing of IFNb in SPMS be limited to those patients who have had disabling relapses in the last 2 years. These conclusions should be reviewed when the full results of all four trials have been published. PMID- 12559514 TI - Interferon beta-1a in primary progressive multiple sclerosis. AB - There is currently no disease-modifying treatment proven to be of efficacy in primary progressive multiple sclerosis (PPMS). However, a number of therapeutic trials have recently been specifically designed for this group. These include a randomised controlled trial of interferon beta-1a which is discussed here. It is hoped that therapeutics in primary progressive multiple sclerosis will continue to expand and effective therapeutic agents will be developed. PMID- 12559513 TI - Low interleukin-10 production is associated with higher disability and MRI lesion load in secondary progressive multiple sclerosis. AB - Abnormalities in T-cell-derived cytokine production are a well-known phenomenon in multiple sclerosis (MS). An association between disability and the production of interferon gamma has been demonstrated recently. The present study investigated associations between disability, cytokine production in stimulated blood lymphocytes and magnetic resonance imaging data in 37 patients with the secondary progressive course in the stable phase of the disease. Patients with high interleukin-10 (IL-10) production had significantly lower disability scores (p=0.009) and lower T2 lesion load (p=0.03). Interleukin-10 might not only play a role in the pathological process of multiple sclerosis but has an impact on disease outcome as well. PMID- 12559515 TI - Gene therapy delivery of endostatin enhances the treatment efficacy of radiation. AB - BACKGROUND AND PURPOSE: To evaluate whether sustained expression of mouse endostatin by adeno-associated virus (AAV)-mediated gene transfer can enhance the treatment efficacy of ionizing radiation. MATERIALS AND METHODS: Mouse endostatin was cloned into recombinant AAV (rAAV) under the control of CMV beta-actin promoter. Recombinant mouse endostatin expressed via AAV gene transfer was tested for biological activity in endothelial cells. The impact of elevated serum levels of endostatin on tumor-induced angiogenesis was evaluated using an in vivo angiogenesis assay. The anti-tumor efficacy of combining rAAV-mediated endostatin delivery with radiation was evaluated in a human colorectal tumor model (HT29). RESULTS: Recombinant mouse endostatin expressed through an AAV vector (rAAV mEndo) inhibited endothelial cell proliferation (by 40-45%) and migration (by 22 33%). Intramuscular injection of rAAV-mEndo (1x10(9) i.u.) led to a sustained serum endostatin level of approximately 500 ng/ml. Compared to control animals this endostatin level was sufficient to inhibit tumor cell-induced vessel formation (37 vs. 28.5, P<0.05) and delay the growth of HT29 xenografts (time from 200 to 1,000 mm(3), 21 vs. 34.5 days, P<0.05). When combined with ionizing radiation, elevated serum endostatin levels significantly enhanced the time for tumors to grow from 200 to 1,000 mm(3) (radiation, 34 days; endostatin plus radiation, 50 days, P<0.05). CONCLUSION: The delivery of endostatin via rAAV vectors may provide an effective means of enhancing the anti-tumor efficacy of radiation therapy. PMID- 12559516 TI - Intensity modulation in radiotherapy: photons versus protons in the paranasal sinus. AB - PURPOSE: The purpose of this study is to investigate whether successive tightening of normal tissue constraints on an intensity modulated X-ray therapy plan might be able to improve it to the point of clinical comparability with the corresponding intensity modulated proton therapy plan. MATERIALS AND METHODS: Photon and proton intensity modulated plans were calculated for a paranasal sinus case using nominal dose constraints. Additional photon plans were then calculated in an effort to match the dose-volume histograms of the critical structures to those of the proton plan. RESULTS: On reducing the low dose contribution to both orbits in the photon plan by tightening the constraints on these structures, an increased dose heterogeneity across the target resulted. When all critical structures were more strictly constrained, target dose homogeneity and conformity was further compromised. An increased integral dose to the non-critical normal tissues was observed for the photon plans as dose was progressively removed from the critical structures. CONCLUSIONS: Both modalities were found to provide comparable target volume conformation and sparing of critical structures, when the nominal dose constraints were applied. However, the use of intensity modulated protons provided the only method by which critical structures could be spared at all dose levels, whilst simultaneously providing acceptable dose homogeneity within the target volume. PMID- 12559517 TI - Intensity modulated radiotherapy treatment planning for dynamic multileaf collimator delivery: influence of different parameters on dose distributions. AB - BACKGROUND AND PURPOSE: This is an investigation of a dose-based conjugated gradient optimization method (implemented in the CadPlan/Helios system) applied for head and neck tumours. Optimized field fluence distributions are created and transformed into dynamic multileaf collimator (MLC) movements. The aim was to gain knowledge about the influence of different parameters on the dose distribution and how to use the optimization algorithm in an optimum way. MATERIAL AND METHODS: Parameters such as the number of beams, collimator angle and constraints and weight factors have been investigated. Dose escalation to the target volume, the target volume in the build-up region and the way of prescribing the target dose were also investigated. The dose distributions were mainly analysed with physical parameters. RESULTS AND CONCLUSIONS: The optimization algorithm is well suited to create clinical Intensity modulated radiation therapy (IMRT) treatment plans for head and neck tumours even when the target volume is situated in the build-up region. The number of beams is a critical parameter and has a great influence on the dose distribution. The choice of collimator angles is not a critical parameter. The constraints and weight factors have a great influence on the dose distribution and varying these could easily control priorities regarding dose to the target volume or to the surrounding critical organs. Because of dose variations inside the target volume, prescribing to, normalizing to and reporting the mean dose in the target volume for IMRT treatment plans is preferable to the absorbed dose at a point, for example the isocentre point. PMID- 12559518 TI - Comparative analysis of intensity modulation inverse planning modules of three commercial treatment planning systems applied to head and neck tumour model. AB - BACKGROUND AND PURPOSE: Three commercial treatment planning modules for intensity modulated radiation therapy (IMRT) Inverse Planning, MDS-Nordion Helax-TMS, Varian Cadplan-Helios, and CMS Focus, were compared in an attempt to determine potential application limits or dosimetric differences among various optimisation algorithms. MATERIALS AND METHODS: A comparative analysis of intensity modulated dose distributions was conducted at planning level on a group of four patients presenting advanced head and neck cancers. In the study, we analysed primarily the static 'step and shoot' multileaf implementation of modulation realisation with some investigation, on the Cadplan-Helios implementation of the 'sliding window', the Varian dynamic approach to IMRT delivery. The whole study was carried out using the inverse planning tools implemented by vendors fully optimising each plan to obtain the best dosimetry given some general plan objectives. To achieve adequate target coverage, optimisation was carried out on Helax-TMS and CMS Focus adding extra margins of 5 or 6mm to the planning target volume (PTV). Beam arrangements were set with five and nine equally spaced fields. The study was conducted with two complexity levels. At the first level, dose-volume constraints were applied only to the target volume and to the spinal cord, while parotid glands were added at the second level. The relative values of dose distributions and dose-volume histograms were compared, together with an estimate of the biological implications in terms of Equivalent Uniform Dose to the target. In the Cadplan-Helios system also the dosimetric implications of the number of intensity levels selected for the discretisation of the fluence matrix were investigated. RESULTS: With the application of common planning strategies and the proper consideration of treatment planning system (TPS) specific features (e.g. the PTV margin problem), no substantial differences among the three algorithms were demonstrated at the first level for PTV and spinal cord. At the second level of the study differences were outlined for Helax-TMS, where sub optimal results were obtained with the 5-field geometry. Mainly due to the differences in optimisation volumes, Cadplan-Helios presented significant better sparing of healthy tissue around the PTV, in terms of mean dose to healthy tissue and Irradiated Volume at 50% dose level. Finally, to achieve dosimetrically acceptable and stable results on target, a minimum of eight intensity levels should be applied for the multileaf collimator (MLC) segmentation, giving an average of 1.5 segments per field and per intensity level. CONCLUSIONS: Results obtained for the three IMRT TPS show in first instance that the optimisation algorithms analysed, as well as the conversion from computed fluences to multileaf sequences implemented in the planning systems can produce substantially equivalent dose plans (for target coverage and organs at risk sparing) if planning is performed with common strategies and once a strong understanding of each system feature is achieved. Secondly, a limited number of dose levels (about eight) is adequate at planning level. PMID- 12559519 TI - Le Fort I osteotomy and low-dose rate Ir192 brachytherapy for treatment of recurrent nasopharyngeal tumours. AB - BACKGROUND: Treatment of recurrent nasopharyngeal carcinoma is a difficult clinical problem. External beam re-irradiation is associated with a long-term cure in a proportion of cases but this may be associated with severe radiation injury. METHODS: Eighteen patients with post-nasal space tumours were treated between 1986 and 2001 with surgical excision and nasopharyngeal brachytherapy via a Le Fort I osteotomy approach. Low-dose rate (LDR) and high-dose rate (HDR) brachytherapy was used. Data was prospectively collected. Local control and overall survival were measured. Acute and late complications were assessed using the RTOG system. RESULTS: The overall survival was 67% at 2 years and 33.5% at 5 years. The corresponding local control rates were 42 and 31.5%, respectively. The T stage at relapse was a significant prognostic factor for local control (P=0.004) but not overall survival. Acute toxicity was mild. RTOG grade >/=3 late complications occurred in 40% of patients treated with the HDR, and 0% treated with LDR. CONCLUSIONS: The results of the Le Fort osteotomy, tumour debulking and post-operative brachytherapy gives local control rates similar to those achieved with wide-field re-irradiation. Complication rates are acceptable and are lower than that reported with other methods of radiation therapy. The surgical technique was well tolerated. HDR brachytherapy with this technique had a high complication rate. This approach is a viable option in the treatment of recurrent nasopharyngeal tumours. PMID- 12559520 TI - Analysis of mandibular dose distribution in radiotherapy for oropharyngeal cancer: dosimetric and clinical results in 18 patients. AB - BACKGROUND AND PURPOSE: The relationship between the radiation dose and the risk of the osteoradionecrosis is well known. However, the dose to the mandible is not routinely assessed in the radiotherapy for head and neck cancer. The aim of our study was to analyze the mandibular dose distribution in the patients administered curative radiotherapy for squamous cell carcinoma of the oropharynx. Moreover, the clinical results have been analyzed. MATERIAL AND METHODS: We examined the clinical records and treatment plans in 18 patients treated with bifractionated radiotherapy for stage II-IV oropharyngeal cancer. In 17 patients, the total radiotherapy dose prescribed in the International Committee of Radiation Units and Measurements (ICRU) reference point was 74.4 Gy administered in 62 fractions (1.2 Gy twice daily with 6h interfraction interval) and one patient received a dose of 75.6 Gy. The whole dose to the mandibular-orophryngeal region was delivered with 6 MV photons. The mandible was contoured manually on computed tomographic scans and the point doses at the both mandibular condyles, ascending ramus, mental symphysis, molar and retromolar regions were assessed. Moreover, the cumulative dose-volume histograms (DHVs) were evaluated. The median follow-up period for alive patients is 30 months (range, 21-44+ months). RESULTS: Tumor remission was observed in 17 patients: in 11 cases, complete remission was achieved and in six cases, only partial remission was possible. One patient was lost to follow-up before the first response evaluation. The median survival for all patients is 22 months (range, 3-44+ months). Ten patients are alive and seven died. In six cases, the cause of death was head and neck tumor and in one died due to pancreatic cancer (second primary). No late bone post-radiation complication was seen. The highest radiotherapy doses were observed in the retromolar regions. The mean percentage doses at the right and left retromolar regions were 101.3+/-3.8% (range, 90.2-109.1%) and 101.7+/-2.5% (range, 95.2 105.8%), respectively. Lower doses were seen in ascending ramus (the mean percentage doses at right and left ascending ramus were 97.3+/-8.5% and 97.8+/ 7.6%, respectively), the molar regions (the mean percentage doses at right and left molar regions were 86.0+/-13.5% and 88.1+/-12.9%, respectively), and at the mandibular condyles (the mean percentage doses at the right and left mandibular condyles were 72.6+/-18% and 77.0+/-16.5%, respectively). The volume of the mandible ranged from 60.1 to 110.1cm(3) (a mean of of 82.3 cm(3)). In all patients, the maximum dose absorbed in the mandible was higher than the dose prescribed in the ICRU point and the mean maximum dose absorbed in the mandible was 105.7+/-2.1% (range, 102.4-110.6%). The percentage of mandibular volume receiving a dose higher than prescribed was 28.6+/-14.9% (range 10.2-58.1%). The area underlying the DVH curve, the maximum mandibular doses and the retromolar doses did not appear to statistically depend on use of wedge or mandibular volume. CONCLUSIONS: Radiotherapy for oropharyngeal cancer is associated with high doses to the retromolar mandibular regions (the dose can be higher than prescribed in the ICRU reference point), ascending ramus and molar regions. Lower doses are absorbed at the condyles and mental symphysis. The single dose point (for example, the ICRU reference point) could be not used as a representative for the mandibular dose. In our small series of patients treated with hyperfractionated irradiation, these dose heterogeneities were not correlated to the patient- and treatment-related factors and are not related to the increased risk of late bone complications. The clinical relevance of mandibular dose distribution remains to be established in larger series of patients treated with conventionally and unconventionally fractionated irradiation. PMID- 12559521 TI - Neck dissection following radiochemotherapy of advanced head and neck cancer--for selected cases only? AB - PURPOSE: To address the value of neck dissection (ND) in patients with advanced head and neck cancer following primary radiochemotherapy and to specifically analyse its impact on locoregional tumour control, survival and toxicity. PATIENTS AND METHODS: Between 1987 and 1997 (9,335), a total of 142 patients (pts) were treated by primary radiochemotherapy (RCT) according to prospective protocols. There were 64 pts with involvement of the hypopharynx, 57 pts with oropharyngeal and 21 with oral cavity carcinoma. UICC (1997) stages included: 16 pts in stage III, 113 pts in stage IV A, 13 pts in stage IV B. All pts received platin-based RCT up to a median total dose of 70 Gy (range, 60-72 Gy). Six weeks after RCT, pts with complete response of the primary tumour (N=97) were offered a uni- or bilateral ND depending on the initially diagnosed nodal disease as part of a strict institutional policy. Fifty-six pts consented to ND and 41 refused. These two groups were analysed in terms of characteristics, local and regional tumour control, survival and long-term side effects. Median follow-up was 37 months (range, 22-124 months). RESULTS: Among the 56 pts receiving ND, a total of 13 (23%) was found to have residual tumour in the neck specimen. The rates of positive histology according to clinical N category after RCT were: yN0 (2/22[9%]), yN1 (2/10[20%]), yN2a-b (2/10[20%), yN2c-3 (7/14[54%]). Five-year overall survival and disease-specific survival rates for pts with ND were 44 and 55%, for pts without ND 42 and 47%, respectively (P=0.9). No difference was seen for long-term local and regional control between the two patient groups. Comparing the group of patients with and without ND, a trend towards higher subjective morbidity of grade 3 and 4 (LENT-SOMA), i.e. pain recording (24% vs. 17%), dysphagia (48% vs. 35%) and hoarseness (20% vs. 9%) was evident in patients with ND. CONCLUSION: No clear evidence for routine clinical use of ND after RCT in advanced head and neck tumours can be derived from these data. ND may be contemplated in selected cases with multiple residual nodes only. PMID- 12559522 TI - Treatment of head and neck cancer with CHART and nimorazole: phase II study. AB - BACKGROUND AND PURPOSE: Causes of failure of radiotherapy in squamous cell carcinoma of the head and neck probably include repopulation and hypoxia. Very accelerated schedules such as continuous hyperfractionated accelerated radiation therapy (CHART) overcome the repopulation problem but allow limited time for reoxygenation, so a hypoxic-cell sensitizer may be especially beneficial. Nimorazole is the only such agent to have shown a significant effect in a randomized controlled trial in head and neck cancer. Accordingly we studied the combination of CHART and nimorazole. METHODS: Sixty-one patients with advanced stage III (21) or IV (40) squamous cell carcinoma of the head and neck unsuitable for surgery were treated in a phase II study of the combination. The radiation dose was 56.75 Gy in 36 fractions in 12 consecutive days. Nimorazole was administered orally or enterally 90 min before radiotherapy at a dose of 1.2, 0.9, and 0.6 g/m(2) with the first, second and third daily fractions, respectively. This dosage consistently yielded plasma concentrations above 30 microg/ml. RESULTS: All the patients have been followed for a minimum of 2 years since treatment. Loco-regional control at 2 years is 55%, stage III 62% and stage IV 50%. Normal tissue effects were the same as those previously seen with CHART, except for a possible slight increase in acute skin reaction. Nimorazole toxicity was somewhat greater than with once daily administration in previous studies. Grade 3 nausea or vomiting occurred in 22% of patients. Two patients developed grade 1 peripheral neuropathy, and one patient died during treatment of encephalopathy, which was probably an idiosyncratic reaction to the drug. CONCLUSIONS: Local control rates are higher than those previously seen with CHART, suggesting a positive effect of nimorazole. Further studies of hypoxia modifying agents with accelerated radiotherapy are warranted, and nimorazole is the simplest of these. PMID- 12559523 TI - Influence of haemoglobin concentration and peripheral muscle pO2 on tumour oxygenation in advanced head and neck tumours. AB - Haemoglobin concentrations and tumour-pO(2) were evaluated pre-therapy in 30 patients with head and neck cancers. Anterior tibialis muscle-pO(2) was additionally measured in 16 of these patients. Tumour-pO(2) was lower in the most anaemic patients (P=0.032) and correlated with muscle-pO(2) (r=0.809, P<0.001). These results suggest that haemoglobin concentration influences tumour oxygenation. PMID- 12559524 TI - Portal imaging to assess set-up errors, tumor motion and tumor shrinkage during conformal radiotherapy of non-small cell lung cancer. AB - PURPOSE: To investigate patient set-up, tumor movement and shrinkage during 3D conformal radiotherapy for non-small cell lung cancer. MATERIALS AND METHODS: In 97 patients, electronic portal images (EPIs) were acquired and corrected for set up using an off-line correction protocol based on a shrinking action level. For 25 selected patients, the orthogonal EPIs (taken at random points in the breathing cycle) throughout the 6-7 week course of treatment were assessed to establish the tumor position in each image using both an overlay and a delineation technique. The range of movement in each direction was calculated. The position of the tumor in the digitally reconstructed radiograph (DRR) was compared to the average position of the lesion in the EPIs. In addition, tumor shrinkage was assessed. RESULTS: The mean overall set-up errors after correction were 0, 0.6 and 0.2 mm in the x (left-right), y (cranial-caudal) and z (anterior posterior) directions, respectively. After correction, the standard deviations (SDs) of systematic errors were 1.4, 1.5 and 1.3 mm and the SDs of random errors were 2.9, 3.1 and 2.0 mm in the x-, y- and z-directions, respectively. Without correction, 41% of patients had a set-up error of more than 5 mm vector length, but with the set-up correction protocol this percentage was reduced to 1%. The mean amplitude of tumor motion was 7.3 (SD 2.7), 12.5 (SD 7.3) and 9.4 mm (SD 5.2) in the x-, y- and z-directions, respectively. Tumor motion was greatest in the y-direction and in particular for lower lobe tumors. In 40% of the patients, the projected area of the tumor regressed by more than 20% during treatment in at least one projection. In 16 patients it was possible to define the position of the center of the tumor in the DRR. There was a mean difference of 6 mm vector length between the tumor position in the DRR and the average position in the portal images. CONCLUSIONS: The application of the correction protocol resulted in a significant improvement in the set-up accuracy. There was wide variation in the observed tumor motion with more movement of lower lobe lesions. Tumor shrinkage was observed. The position of the tumor on the planning CT scan did not always coincide with the average position as measured during treatment. PMID- 12559525 TI - Analysis of dose distribution in the 'Rind'--a volume outside the PTV--in 3 dimensional conformal radiation therapy of non-small cell lung cancer. AB - BACKGROUND AND PURPOSE: Appropriate planning target volume (PTV) definition is critical for local disease eradication in the treatment of non-small cell lung cancer (NSCLC). When margins are added to the gross tumour volume (GTV) in the standard way, the PTV formed may be too large to facilitate dose escalation due to normal tissue tolerance. To increase the feasibility of dose escalation with 3 dimensional conformal radiotherapy (3DCRT), this study examines an alternative method for the formation of the PTV in NSCLC. This strategy is based on the reduced probability of tumour cells from the GTV outwards and on the associated lower dose requirements to eradicate such subclinical disease. MATERIALS AND METHODS: 3DCRT plans were generated from the CT scans of 15 patients with NSCLC (stages Ib to IIIb). Each PTV was formed by adding a margin for geometric uncertainties directly onto the GTV. The success of this approach is dependent on the volume immediately outside this smaller PTV, the Rind volume, receiving 50 Gy, the minimum dose requirement that is considered sufficient for eradication of the reduced tumour cell density in this volume. While optimizing the treatment plans for each PTV to 70 Gy, the dose distribution in the Rind volume, and the factors affecting it, were assessed. RESULTS: One hundred percent of each PTV received a minimum of 95% of the prescribed dose. The percentage of the Rind volume receiving 50 Gy or more (V50) had a median value of 94%. The minimum dose in this volume, however, ranged from 5.6 to 32.1 Gy. The V50 was highest for apical tumours (96.1%) and lowest for peripheral tumours (86%) and correlated positively with the size of the PTV (Kendall's rank correlation (Kt)=+0.3, P=0.05) and the number of beams used (Kt=+0.3, P=0.03) but not with the conformity index. The average volume outside the Rind which still received >/=50 Gy (the Wasted 50 Gy) increased significantly with the V50 of the Rind volume and was inversely proportional to the Rind <50 Gy, correlating significantly with the dose to the organs at risk. CONCLUSIONS: Using this strategy with standard 3DCRT, all PTVs were irradiated to the required dose with this approach, but none of the corresponding Rind volumes had an acceptable dose distribution. The addition of dual volume planning or the use of intensity modulated radiation therapy may achieve an appropriate dose distribution in the Rind volume while not increasing the dose to the organs at risk and may thereby facilitate dose escalation. PMID- 12559526 TI - Partial volume rat lung irradiation; assessment of early DNA damage in different lung regions and effect of radical scavengers. AB - PURPOSE: These studies were designed to examine radiation-induced in-field and out-of-field DNA damage in rat lung as a function of dose and various volumes of irradiation. They also determined whether superoxide dismutase (SOD) and nitro-L arginine methyl ester (L-NAME) protected against this damage. METHODS AND MATERIALS: The whole lung, or various volumes of the lower or upper lungs of Sprague-Dawley rats were exposed to doses up to 20 Gy of 60Co gamma rays. Radiation-induced DNA damage was quantified in fibroblasts obtained at 18 h after irradiation from both irradiated and shielded lung regions using a micronucleus assay. The radioprotective role of SOD (CuZnSOD: 10 mg/kg body weight; MnSOD: 50 100mg/kg body weight) and L-NAME (0.2 mg/kg body weight.) in vivo was determined by injecting them into rats 30 min before or immediately after a dose of 10 Gy. RESULTS: Micronucleus formation was approximately linear with dose up to 15 Gy. When 70% of the lung volume was irradiated with 10 Gy, irradiated lower lung gave similar numbers of micronuclei (MN)/binucleate cell (BN) to that observed following whole lung irradiation (0.91 MN/BN), whereas the irradiated upper lung gave only 0.66 MN/BN. Following lower lung irradiation, the shielded upper lung (30% of lung volume) showed substantial (out-of-field) damage (0.43 MN/BN). When 30% of the lung was given 10 Gy, irradiated upper or lower lung showed similar amounts of in-field damage (0.43 MN/BN) but this was smaller than that seen following irradiation of 70% of the lung volume. For 30% lower lung irradiation, the shielded upper lung showed only a small out-of-field effect (0.1 MN/BN). For both volumes of irradiation there was a similar or smaller effect in the shielded lower lung after upper lung irradiation. Injection of SOD before or L-NAME after 10 Gy to the lower 70% lung volume resulted in a reduction in DNA damage both in field and out-of-field but the percentage was much greater for out-of-field damage (50-60%) than for in-field damage (10-30%). Following whole lung irradiation (10 Gy) significantly greater DNA damage was observed in fibroblasts from the left lung than from the right lung (0.93 MN/BN vs. 0.82 MN/BN). Following whole lung irradiation there was no significant difference in DNA damage observed in fibroblasts from the lower lung and the upper lung. CONCLUSIONS: With partial lung irradiation the lower lung sustains more in-field DNA damage following irradiation than the upper lung, whereas out-of-field effects are observed primarily in the upper lung (i.e. following lower lung irradiation). Following whole lung irradiation the left lung sustains more damage than the right lung but there is no difference between the upper and lower lung. The protective effects of SOD and L-NAME suggest that inflammatory cytokines induced by the irradiation may be involved in the initiation of a reaction resulting in the production of reactive oxyradicals and nitric oxide that cause indirect DNA damage both in and out of the radiation field. PMID- 12559527 TI - Comparison of medical therapy, pacing and defibrillation in heart failure (COMPANION) trial terminated early; combined biventricular pacemaker defibrillators reduce all-cause mortality and hospitalization. AB - The COMPANION trial was terminated prematurely after recruiting 1600 patients, as initial results clearly and for the first time demonstrated survival benefit of resynchronisation and combined device (biventricular pacemaker with defibrillator capacity) therapy in heart failure. The combined primary end-points of all-cause mortality and all-cause hospitalisation in patients with dilated cardiomyopathy and heart failure of poor functional class was reduced by 20% with cardiac resynchronisation therapy (CRT) with or without defibrillator capacity. More importantly, mortality was reduced by 40% (from 19% to 11%) in patients implanted with combined devices, while CRT alone gave an intermediate mortality of 15%. PMID- 12559528 TI - Oral loading single dose flecainide for pharmacological cardioversion of recent onset atrial fibrillation. AB - The efficacy and safety of the single oral loading dose of flecainide for cardioversion of recent-onset atrial fibrillation was examined by reviewing the trials on the subject identified through a comprehensive literature search. Most of the trials used a single dose of 300 mg for oral loading. The success rate ranged from 57 to 68% at 2-4 h and 75 to 91% at 8 h after drug administration. The conversion time ranged from 110+/-82 to 190+/-147 min, depending on the duration of observation after drug administration, which in most trials was of 8 h. Single oral loading regimen of flecainide was significantly more efficacious than placebo, and was as efficacious as the single oral loading regimen of propafenone. Both the single oral loading and the intravenous loading regimens of flecainide were equally efficacious but the intravenous regimen resulted in an earlier conversion. Adverse effects reported were mild non-cardiac side effects, reversible QRS complex widening, transient arrhythmias and left ventricular decompensation. The transient arrhythmias were chiefly at the time of conversion and included appearance of atrial flutter and sinus pauses. No life-threatening ventricular arrhythmia or death was reported. The single dose oral loading regimen of flecainide appears to be effective for cardioversion of recent-onset atrial fibrillation with a relatively rapid effect within 2-4 h, and is free of serious complications in patients without structural heart disease. Patients with substantial structural heart disease were excluded from most of the trials. PMID- 12559529 TI - Assessment of left ventricular systolic and diastolic functions in children with merosin-positive congenital muscular dystrophy. AB - Cardiopathy is an expected finding in X-linked Duchenne and Becker muscular dystrophies. This holds true for some other forms such as autosomal recessive limb-girdle dystrophies. However, data on early-onset and usually severe congenital muscular dystrophies are limited. The purpose of this study was to investigate the presence of cardiac involvement in children with merosin-positive congenital muscular dystrophy. A total of 42 patients and 22 healthy subjects were evaluated by M-mode, 2D, and Doppler echocardiography. Cardiac anatomy, left ventricular dimensions, wall thickness and systolic and diastolic functions were investigated in patients and compared with those of healthy control subjects. Mean left ventricular ejection fraction and shortening fraction were significantly lower in the patient group (P<0.05 and P<0.001, respectively) and in three patients ejection fraction was below 55%. Although some impairments in left ventricular inflow indexes which were suggestive of left ventricular diastolic dysfunction were detected in patients with merosin-positive congenital muscular dystrophy they were not statistically significant. Our results suggest that left ventricular systolic abnormalities may occur in children with merosin positive congenital muscular dystrophy. PMID- 12559531 TI - Ought dialysis patients with atrial fibrillation be treated with oral anticoagulants? AB - BACKGROUND: Dialysis patients with atrial fibrillation have an increased thrombolic risk. Dicoumarin anticoagulant therapy is often considered contra indicated in chronic renal insufficiency in which the risk of haemorrhage, though not defined, is perceived to be high. We assessed haemorrhage complications in dialysis patients receiving dicoumarin anticoagulant therapy to establish whether the haemorrhage risk justifies the contra-indication of anticoagulant therapy in patients with atrial fibrillation. PATIENTS AND METHODS: Over a period of a decade in our dialysis centre, 29 patients receiving anticoagulant therapy over a protracted period presented haemorrhage complications. These were classified with respect to severity and location and compared with 211 patients not receiving anticoagulant therapy. The relative risk of haemorrhage was calculated and was compared to risk of thrombo-embolism in dialysis patients with atrial fibrillation. RESULTS: Of the 29 patients, nine had 13 episodes of haemorrhage complications (26 episodes/100 patient-years). None was fatal, nor intra-cranial nor with serious clinical sequelae. In the group without anticoagulants, 29 patients had 39 haemorrhage complications (11 episodes/100 patient-years); four (10.2%) intra-cranial and all fatal. The relative risk of bleeding with anticoagulant therapy was 2.36 (95% confidence interval=1.19-4.27). CONCLUSIONS: (1) Dialysis patients with anticoagulant therapy presented with a higher risk of haemorrhage; (2) the relative risk of bleeding was double that of the dialysis population without anticoagulant therapy; (3) despite the high risk of haemorrhage that we observed, the high risk of thrombo-embolism and the attendant serious sequelae to which dialysis patients with atrial fibrillation are predisposed indicates that oral anticoagulation therapy ought not to be considered automatically contra-indicated in this patient group but that an exhaustive evaluation of the risk-benefit needs to be conducted on an individual patient basis. PMID- 12559533 TI - Comparison of clopidogrel versus ticlopidine for prevention of minor myocardial injury after elective coronary stenting. AB - We searched a randomized, double-blinded, prospective study that compared the effectiveness of clopidogrel versus ticlopidine for prevention of minor myocardial injury (MMI) and major clinical events (MCEs) after elective coronary stenting. A total of 158 consecutive patients (98 male, 60 female patients with a mean age of 59.3+/-5.4 years) were divided into two arms based on treatment with thienopyridines: group I, clopidogrel 1 x 300 mg as a loading dose, and 1 x 75 mg per day thereafter, group II, ticlopidine 2 x 250 mg daily. Both thienopyridines were started on the same day as stent placement. Cardiac troponin T (cTnT) was measured immediately before and 12 h after the procedures. All patients were followed-up during the hospital stay (6+/-2 days) with respect to MMI and MCEs. The increase frequency and the amount of cTnT level in group I was found significantly lower compared with group II (5 vs.15; P<0.01; 0.38+/-0.11 vs. 0.44+/-0.12 ng/ml; P<0.001, respectively). Patients with elevated cTnT levels more likely to have C type lesion (P<0.004). Though there was a trend toward increased major clinical events rate in group II than those of group I, the statistical difference was not different (4 vs. 1.3%; P>0.05). The present study showed that the combination of clopidogrel and aspirin was more effective than the combination of ticlopidine and aspirin in decreasing the rate of MMI. PMID- 12559534 TI - The prevention of minor myocardial injury with ticlopidine pretreatment in patients undergoing elective coronary stenting. AB - The aim of the study was to determine whether ticlopidine treatment prior the coronary stenting would be associated with lower rates of procedure-related minor myocardial injury (MMI) in patients undergoing elective coronary stenting. In this retrospective, nonrandomized, uncontrolled study, a total of 153 consecutive patients with a mean age of 63.4+/-8.9 years were divided into two groups based on the duration of ticlopidine treatment: group I (n=81), ticlopidine >/=3 days before the procedure, group II (n=72), on the same day as stent placement. Cardiac troponin T (cTnT) was measured immediately before and 12 h after the procedures. All patients were followed-up during the hospital stay with respect to MMI and major clinical events (MCE). The increase frequency and the amount of cTnT level in group I was found to be significantly lower compared with group II (4 vs. 13; P<0.01, and 0.35+/-0.06 vs. 0.52+/-0.11 ng/ml; P<0.01, respectively). In general, patients with elevated cTnT levels are more likely to have C type lesion and multivessel procedure than those of normal cTnT level (41 vs.10%; P<0.002 and 47 vs. 17%; P<0.009, respectively). Though there was a trend toward increased MCE rates in group II than that of group I, this did not reached statistical significance (3 vs.1; P=NS). The present study shows that an anti platelet treatment with ticlopidine prior the coronary stenting of adequate duration to allow for the development of maximal inhibition is associated with a markedly decreased incidence of procedure-related MMI. Therefore, ticlopidine pretreatment may be a cost alternative for the prevention of platelet-rich microembolism in patients undergoing elective coronary stenting. PMID- 12559535 TI - Coronary calcium detected by digital cinefluoroscopy and coronary artery disease in patients undergoing coronary arteriography: effects of age and sex. AB - BACKGROUND: Coronary artery calcium, detected non-invasively, correlates well with angiographically documented coronary artery disease (CAD). This study was conducted to evaluate the diagnostic efficacy of coronary artery calcium detected by digital cinefluoroscopy for CAD and assess the effects of age and sex on it. METHODS: In 242 patients who underwent coronary angiography, coronary calcium status was determined and related to angiographic findings. RESULTS: Calcium detection had a sensitivity 85%, specificity 52%, positive predictive value 92%, negative predictive value 33% and diagnostic accuracy 81% for significant CAD. There was a better positive predictive value in men (95% vs. 80%) and negative predictive value in women (65% vs. 16%), while a higher sensitivity and diagnostic accuracy was found in older than in younger (90% and 86% vs. 78% and 74%). The sensitivity of the method increased with the number of the diseased vessels. CONCLUSIONS: Coronary calcium can be quite accurately detected by digital cinefluoroscopy. This, however, should be made in the context of sex and age. PMID- 12559536 TI - Effects of verapamil and metoprolol on recovery from atrial electrical remodeling after cardioversion of long-lasting atrial fibrillation. AB - The aim of this prospective, randomized study was to investigate the effect of pretreatment with two different intracellular calcium-lowering drugs (verapamil and metoprolol) on recovery from atrial effective refractory period (AERP) shortening after internal electrical cardioversion (EC) of persistent atrial fibrillation (AF) in patients on amiodarone. Twenty-one patients on amiodarone for at least 30 days were referred to our hospital for internal EC of a persistent AF refractory to external EC. They were randomized to receive only amiodarone (group AMI, n=7), or amiodarone and verapamil 240 mg/day (group VER, n=7), or amiodarone and metoprolol 100 mg/day (group MET, n=7). Left AERP was measured 10 min and 24 h after EC. AERP was also determined in 13 controls. The AERP after 10 min was significantly shorter in group AMI (201 (31) ms, P<0.02) and group MET (203 (34) ms, P<0.03) than in controls (249 (45) ms), but not in group VER (237 (51) ms, P=NS). The AERP after 24 h was still significantly shorter in group AMI (204 (38) ms, P<0.04) than in controls, but not in group MET (225 (52) ms, P=NS) or in group VER (290 (36) ms, P=NS). Pretreatment with amiodarone and verapamil prevents AERP shortening, while pretreatment with amiodarone and metoprolol only accelerated AERP recovery. PMID- 12559537 TI - Aerobic exercise physiology in a professional rugby union team. AB - INTRODUCTION: In professional rugby, different positional roles may require different levels of aerobic fitness. Forward and backline players from a team of elite rugby players were tested to evaluate the differences between the two groups. METHODS: 28 male players, 15 backs and 13 forwards, underwent maximal treadmill cardiopulmonary exercise testing (CPX), lung spirometry, a 3 km timed run, and body fat measurement. RESULTS: Peak oxygen uptake was higher in backs than in forwards (peak VO(2) 48.3+/-2.1 vs. 41.2+/-2.7 ml kg(-1) min(-1), P<0.05) with no significant difference in peak respiratory exchange ratio (1.08+/-0.02 vs. 1.07+/-0.02, P=NS), exercise time (1306+/-39.7 vs. 1217+/-25.1 s, P=NS) or time for 3 km run (667.5+/-14.1 vs. 699.0+/-20.7 s, P=NS). However, the forwards were taller and heavier (height 190.2+/-2.2 vs. 179.5+/-1.3 cm, P<0.001, body mass 104+/-2.4 vs. 86.3+/-1.7 kg, P<0.0001) and had a higher fat content (body fat percentage 12.8+/-0.8 vs. 9.7+/-0.6%, P<0.01) and forced expiratory volume in 1 s (FEV1, 4.9+/-0.1 vs. 4.5+/-0.2 l, P<0.05). There was a significant negative correlation between peak VO(2), 3 km run time (r=-0.45, P<0.05) and weight (r= 0.54, P<0.003) for all subjects. CONCLUSION: Backline players have a higher peak oxygen uptake per kilogram than forwards, although the cardiopulmonary exercise test duration, degree of anaerobic metabolism and 3 km run time are not significantly different. These results could be due to the two groups' different body structure, being shorter, lighter and having a lower percentage body fat. These differences, which are likely to be a result of selection for specific roles in the game, should be taken into account when evaluating aerobic fitness within a rugby team. PMID- 12559538 TI - Hormonal profile in patients with congestive heart failure. AB - BACKGROUND: Recent progress has been made in the understanding of the cellular and molecular mechanisms of growth hormone action and of its effects on cardiac tissue. The aim of this study was to measure growth hormone concentrations, along with various other hormones, in patients with stable chronic congestive heart failure due to idiopathic dilated cardiomyopathy. METHODS: The study included 23 ambulatory men, 51.2+/-9.3 years of age, on standard medical therapy for heart failure due to idiopathic dilated cardiomyopathy. All patients underwent clinical and laboratory evaluations, including echocardiogram, radionuclide ventriculography, right heart catheterization, coronary angiography, and right ventricular endomyocardial biopsy. Serum or plasma concentrations of growth, thyroid, sex and adrenal hormones were measured in all patients and compared with those found in 20 age-matched healthy men. RESULTS: Growth hormone, insulin-like growth factor I, and free testosterone values in patients with idiopathic dilated cardiomyopathy and heart failure were 0.37+/-0.2 ng/ml, 123.7+/-50 ng/ml and 48.6+/-23.8 pmol/l, respectively, versus 0.5+/-0.4 ng/ml (P<0.01), 236.3+/-66.4 ng/ml (P<0.001) and 105+/-17 pmol/l (P<0.01), respectively, in the healthy age matched individuals. All other hormone concentrations were comparable in both groups. CONCLUSIONS: Chronic heart failure due to idiopathic dilated cardiomyopathy is associated with a significant decrease in growth hormone, insulin-like growth factor I, and testosterone concentrations, probably due to chronic disease. PMID- 12559539 TI - Administration of growth hormone to patients with advanced cardiac heart failure: effects upon left ventricular function, exercise capacity, and neurohormonal status. AB - Experimental and clinical studies have shown that the administration of recombinant human growth hormone can improve deteriorated left ventricular function and hemodynamics in patients with heart failure. Herein, we compared the effects of growth hormone versus placebo upon resting left ventricular ejection fraction, exercise capacity and neurohormonal status in patients with advanced heart failure. Nineteen patients with advanced cardiac heart failure (ejection fraction <30%) were studied at baseline and after 8 weeks of treatment with growth hormone (0.03 U/kg per day) or placebo. Primary end points were resting left ventricular ejection fraction, peak oxygen consumption and neurohormonal status, including plasma norepinephrine levels and insulin like growth factor-1 and its binding protein-3. Results are presented as median and interquartile ranges. Patients receiving growth hormone had a significant increase in insulin growth factor-1 plasma levels (median difference growth hormone=83 ng/ml [57-170] versus placebo=-6 ng/ml [-23-6], P<0.05) and its binding protein-3. However, no significant increase in left ventricular ejection fraction after growth hormone treatment (ejection fraction pre=16% [13-18] and post=17% [14-27]) was noticed when compared to placebo (ejection fraction pre=20% [15-24] and post=20% [15 26]). Also, no significant effect of growth hormone treatment was seen on peak oxygen consumption or norepinephrine plasma levels. Although the administration of growth hormone to patients with advanced cardiac heart failure was associated with a significant increase in insulin growth factor-1, there were no significant changes in ejection fraction, exercise capacity and/or neurohormonal status. PMID- 12559540 TI - Low density lipoprotein particles are small in patients with coronary vasospasm. AB - Increased lipid oxidative stress has been recently implicated in the pathogenesis of coronary artery spasm. Small, dense LDL with high susceptibility to oxidation may be linked to the genesis of coronary vasospasm. The relative migratory distance of the predominant densitometric peak of LDL from that of VLDL to that of HDL in a 3% polyacrylamide gel electrophoresis was determined as a measure of LDL particle size in 49 patients with coronary spastic angina (CSA), in 56 patients with stable effort angina and a significant coronary artery stenosis (SEA) and also in 40 control subjects without coronary artery disease (Control). The incidence of detection of small, dense LDL (particle diameter <25.5 nm) or a relative migratory distance above 0.36 was significantly higher in CSA (57%) and also in SEA (39%) than in Control (20%). In SEA, a significantly higher serum level of triglyceride was noted in the subgroup with the small, dense LDL as compared with the subgroup without. In contrast, in CSA, the serum level of triglyceride was not significantly different between the subgroups with and without the small, dense LDL, although significantly lower serum levels of both HDL-cholesterol and alpha-tocopherol were noted in the former. In 16 patients of CSA, the detection of the small, dense LDL was significantly decreased after a >6 month angina-free period (69-->31%). We conclude that patients with coronary spastic angina had smaller LDL particles, associated not with hypertriglyceridemia but low serum levels of both HDL-cholesterol and vitamin E. Dyslipidemia with small, dense LDL may be related to the genesis of coronary vasospasm. PMID- 12559541 TI - The cow and the coronary: epidemiology, biochemistry and immunology. AB - BACKGROUND: Certified death rates from coronary heart disease (CHD) are positively correlated country-by-country with milk consumption, particularly with that of the non-fat portion of milk. CHD death is also associated with circulating antibodies against milk fat-globule membrane (MFGM), raising the possibility that milk intake might be a specific risk factor for CHD. We studied the epidemiology and immunology of milk to see if the association is causal. METHODS: We plotted the intake of various foods country-by-country against CHD death rates to see if they were correlated in space and/or in time. We prepared fluorescein-tagged human antibodies against bovine MFGM to see if they showed any auto-reactivity against human tissue. RESULTS: Milk showed a positive correlation with CHD death rates in both space and time (r>0.9 in some cases). Beef was not correlated and cheese was negatively correlated, though not strongly. Wine was strongly negatively correlated. Human anti-bovine MFGM antibodies bind to human large granular lymphocytes and also to human platelets, causing aggregation. CONCLUSIONS: We suggest that non-fat aspects of milk, particularly the Ca/Mg ratio, lactose and MFGM antigens, have specific coronary atherogenic effects, both biochemical and immunological, and the simultaneous attack from these three directions may explain why this foodstuff has such a strong effect. Wine appears to be an antidote for the harmful effects of milk. PMID- 12559542 TI - Changes in QT interval with exercise in elite male rowers and controls. AB - BACKGROUND: QT interval prolongation occurs at rest and during exercise in pathological left ventricular hypertrophy. However, athletes with physiological hypertrophy have normal QT at rest. The aim of this study was to compare the effect of exercise on QT in athletes with echocardiographic left ventricular hypertrophy and normal controls, and explore differences in their response. METHODS: Elite male rowers (n=15) with echocardiographic left ventricular hypertrophy, and normal volunteers (n=15) underwent 15 min of a Bruce protocol treadmill test. Electrocardiograms (ECGs) were recorded during each stage and every minute during recovery for 3 min. QT was measured at each stage. Corrected QT (QTc) was calculated using Bazett's formula. RESULTS: QT at rest was significantly greater than QT after 3 min of recovery in the controls (0.36+/ 0.02 vs. 0.32+/-0.04 s; P=0.001) but not in the athletes (0.36+/-0.03 vs. 0.34+/ 0.02 s; P=0.05). Regression lines for QT versus heart rate showed a strongly negative correlation in both athletes and controls (y=0.463-0.0013x (r=0.91; P<0.0001) and y=0.461-0.0013x (r=0.93; P<0.0001), respectively), but greater individual homogeneity in the athletes. CONCLUSIONS: training-induced hypertrophy does not affect the heart rate/QT relationship. The more rapid recovery in QT and homogeneity of the heart rate/QT relationship in athletes compared to controls is likely to be a benign effect of myocardial fitness, but it is hypothesised that it may contribute to arrhythmias in the unfit individual after vigorous exertion. PMID- 12559543 TI - Cardiac troponin T and myocardial injury during routine cardiac catheterisation in children. AB - BACKGROUND: This study aims to investigate whether intracardiac catheterization produces myocardial damage on pediatric heart. METHODS: Five blood samples were collected (basal, immediate post procedure, at 4, 12 and 24 h after the procedure) for troponin T and creatine kinase MB (CKMB) from 48 consecutive patients (age: 5.34+/-6.03 years). The effect of age, duration of procedure, pulmonary hypertension, cyanosis, and medication taken for congestive heart failure on the levels of troponin T and CKMB were sought. RESULTS: The increase in CKMB (basal CKMB: 3.93+/-3.70 ng/ml; peak CKMB: 8.68+/-10.89 ng/ml; P<0.0001) and troponin levels (basal troponin: 0.002+/-0.003 ng/ml; peak troponin: 0.11+/ 0.23 ng/ml; P<0.0001) over time was significant in the study group. Additionally younger patients (25 mmHg), longer procedure time (>30 min), and patients taking anti-congestive heart failure therapy had significantly higher levels of CKMB and troponin (P>0.05). CONCLUSION: All patients undergoing cardiac catheterization are under risk of myocardial injury, and younger patients with pulmonary hypertension and especially with compensated cardiac failure have increased risk of myocardial damage, and need to be handled carefully. PMID- 12559544 TI - Electrocardiographic criteria for vagotonia-validation with pharmacological parasympathetic blockade in healthy subjects. AB - BACKGROUND: The importance of vagal tone on cardiac function and cardiovascular mortality is well established. Although the presence of an enhanced cardiac vagal tone (CVT) is frequently diagnosed using the 12-lead resting electrocardiogram (ECG) in daily practice, most of the proposed criteria have been determined on an empirical basis. Our objective was to evaluate the effects of pharmacological blockade of the parasympathetic component of the autonomic nervous system on resting ECG tracings. METHODS: Nine healthy young adults (24+/-5 year-old) underwent parasympathetic blockade with atropine sulfate i.v. (0.04 mg kg(-1)) and resting ECGs were obtained before and 15 min thereafter. CVT was assessed by a dimensionless index, which measures the RR interval reduction caused by the vagal withdrawal induced by a 4-s exercise test performed on a cycle ergometer where the subjects pedal as fast as possible with no added resistance. RESULTS: This index was 1.63+/-0.24 and 1.03+/-0.03, before and after atropine, respectively (P<0.0001). Atropine reduced the R-R intervals (P<0.0001), and the amplitude of T-waves in several leads (DII: P=0.03; V4: P=0.04; V5: P=0.03; V6: P=0.01), and abolished the appiculation of T-waves, J-point and ST-segment elevations (P<0.05), and U-waves (P<0.05), which were present in baseline ECG in all subjects in at least two leads. The R-wave amplitude in leads V4, V5, and V6 (all P>/=0.10) was not modified by atropine infusion. CONCLUSION: The duration of the R-R intervals and the amplitude of T-waves in leads DII, V4, V5, and V6, and the presence of T-wave appiculation, U-waves, and elevation of J-point and ST segment should be used to detect enhanced cardiac vagal tone in healthy subjects. PMID- 12559545 TI - Exercise training without ventricular remodeling in patients with moderate to severe left ventricular dysfunction early after acute myocardial infarction. AB - BACKGROUND: The purpose of this study was to determine whether or not patients with moderate to severe left ventricular (LV) dysfunction benefit from exercise training starting early after acute myocardial infarction (AMI) without deteriorating LV remodeling. METHODS: We investigated changes in exercise capacity and LV end-diastolic dimension (LVDd by two-dimensional echocardiography) before and after exercise training in 126 patients after AMI. Patients were divided into three groups according to LV ejection fraction (EF) at the beginning of exercise training: 74 patients with LVEF>/=45% (Group H), 35 patients with 35%40 years (p < 0.001). A cubic function correlated BMAC VUC (x) with chronological age (y): y = 33.7 - 3.29x + 0.072x(2) - 0.0004x(3) (r = 0.48). CONCLUSIONS: Breath markers of OS were significantly increased both in younger and in older subjects, compared to those aged 20-40 years. Increased OS in older subjects was consistent with previous reports, but increased OS in younger subjects aged < 20 years is a new observation; this may be a normal physiological response in youth. PMID- 12559602 TI - Correlation of B-type natriuretic peptide level to 6-min walk test performance in patients with left ventricular systolic dysfunction. AB - BACKGROUND: B-type natriuretic peptide (BNP) is a neurohormone that can be measured in blood and is useful in patients with congestive heart failure (CHF). We compared whole blood BNP concentrations to distance walked during a 6-min walk test in patients with CHF. METHODS: Forty-four patients with CHF underwent a 6 min walk test. The distance walked was compared to the BNP concentration on blood collected prior to the walk test. Patients were followed for 16 +/- 2.8 months after testing. RESULTS: A significant correlation was observed between the BNP concentration and the distance walked (r = -0.47, p < 0.001). One patient without congestion died suddenly. Two patients died of progressive heart failure, and two other patients underwent cardiac transplantation. Each of the latter four patients had high BNP concentrations (median 1080 ng/l) and walked short distances (median 183 m). This study indicates that the BNP concentration in blood correlates inversely with the degree of physical capability of patients with heart failure. CONCLUSIONS: The BNP concentration could be used as an alternative to the 6-min walk test to assess the severity of heart failure. The assay for BNP is non-invasive, inexpensive, and results are available at the bedside or in a heart failure clinic. PMID- 12559603 TI - Differential apolipoprotein(a) isoform expression in heterozygosity is an independent contributor to lipoprotein(a) levels variability. AB - BACKGROUND AND METHODS: Lipoprotein(a) [Lp(a)] levels represent an independent risk factor for cardio- and cerebrovascular diseases. Since lipoprotein(a) levels show a wide variability even in subjects with similar apolipoprotein(a) isoforms, we investigated the contribution of apolipoprotein(a) heterozygosity to lipoprotein(a) variance. Lipoprotein(a) levels, apolipoprotein(a) isoforms identification and expression, and the correlation with other lipo-apolipoprotein parameters have been investigated in 628 subjects >18 years of age. RESULTS: In our study, 246 subjects were found heterozygous for apolipoprotein(a) isoforms. Lipoprotein(a) levels were higher in females. About 40% of the subjects expressed the larger isoform more intensely than the dominant isoform. Lipoprotein(a) was correlated with apolipoprotein(a) dominant isoform size, HDL-cholesterol and smaller apolipoprotein(a) isoform expression rate. Lipoprotein(a) was independently correlated with the smaller apolipoprotein(a) isoform, with its expression rate and with LDL-cholesterol. The inclusion of the smaller apolipoprotein(a) expression rate in a multiple regression model explained at least an additional 4% of the lipoprotein(a) variance after correction for apolipoprotein(a) size. CONCLUSIONS: The smaller isoforms are not always effectively dominant in heterozygosis since 40% of the subjects expressed more the larger isoform. The individual variability of apolipoprotein(a) isoform expression in heterozygosis could explain part of the lipoprotein(a) levels variability. PMID- 12559604 TI - Applicability of a filter paper method to measure blood lead levels in large populations of Chinese children. AB - BACKGROUND: Capillary filter paper can be useful for screening blood lead concentrations in large populations of geographically dispersed Chinese children. METHODS: A total of 159 at 0-6 years of age were enrolled in the study when they came for routine pediatric care. Paired venous and capillary filter paper samples were collected from all subjects. Graphite furnace atomic absorption spectrophotometry was used to measure blood lead concentrations. Comparability of the lead assay methods was assessed. The concordance coefficient between these two methods was 0.87 (p < 0.01). The values of blood lead concentrations measured by the different methods were comparable statistically. The sensitivity of the filter paper method was 85%, 73% and 75%; specificity was 92%, 99% and 100%; the positive predictive value was 86%, 89% and 100%, and negative predictive value was 91%, 98% and 99% at cut-offs of 10, 15 and 20 microg/dl, respectively. The Youden's index was 77, 71 and 75 at those cut-offs. Values obtained via the filter paper method appeared to be relatively stable up to 4 weeks post collection. CONCLUSIONS: The filter paper method for lead determinations has a sufficiently high sensitivity and specificity and correlates well with venous sampling. The ease with which this technique can be performed and screened samples can be sent to a centralized laboratory by regular mail indicates that it is a promising and attractive reference method, particularly in large populations of widely dispersed young children in China and other countries. PMID- 12559605 TI - Arginase in patients with breast cancer. AB - The mean arginase activity in breast cancers (n = 80) was significantly higher than in control tissues and it accounted for 0.31 +/- 0.23 U/g wet tissue and 0.083 +/- 0.061 U/g (P < 0.05), respectively. With the cutoff value of 0.1 U/g wet tissue, raised arginase activity was observed in 74% of tumors. The preoperative arginase activity in blood serum from women with breast cancer was 11.2 +/- 7.9 U/l (n = 115), and it was significantly higher than in 70 healthy controls, where it was 5.7 +/- 2.4 U/l (P < 0.05). With the cutoff value for normal serum arginase activity above 8.0 U/l, the activity was raised in 10% of control individuals, and in 63% of women with breast cancer. The sensitivity and specificity of the arginase test in blood serum were 63% and 60%, respectively. Two isoforms immunologically identical to human kidney arginases (L-arginine amidinohydrolase) were found in both normal and cancerous breast tissues. The level of anionic form was similar in control and cancerous tissues, whereas the cationic isoform predominated in breast cancer. The cationic isoform was the only one present in serum of both ill and healthy women, and its level was higher in patients with breast cancer. Thus, it can be concluded that the cationic isoform is responsible for the increase of arginase activity in serum of patients with breast cancer. PMID- 12559606 TI - Blood extracellular superoxide dismutase levels in hemodialysis patients pre- and post-hemodialysis and its association with lipoprotein lipase mass and free fatty acid. AB - BACKGROUND: Hemodialysis (HD) is a method of chronic therapy for patients with renal failure. Diabetic nephropathy is the most common underlying disease among HD patients in Japan. A characteristic problem associated with this condition is endothelial cell damage. We have been using extracellular superoxide dismutase (EC-SOD) attached to the heparan sulfate on the endothelial cell surface as a marker of vascular damage. METHODS: This study examined the pre- and post-HD levels of lipoprotein lipase (LPL). The increase in free fatty acid (FFA) is affected by a type of cytokine-like substances, which induces insulin resistance and causes abnormal lipid metabolism. RESULTS: Pre-HD blood samples showed highly significant correlations between EC-SOD and LPL mass (r = 0.792) and between EC SOD and FFA (r = 0.675) (p < 0.0001). EC-SOD, LPL mass, and FFA were remarkably high among the patients who had been placed on HD treatment for over 20 years. CONCLUSION: Because EC-SOD and LPL mass represent heparin-binding proteins, these results were considered to reflect severe vascular damage. PMID- 12559607 TI - An enzyme-linked immunosorbent assay for corticosteroid-binding globulin using monoclonal and polyclonal antibodies: decline in CBG following synthetic ACTH. AB - BACKGROUND: Adrenal function is commonly assessed by measuring plasma cortisol following synthetic ACTH (synacthen) challenge. Generally little regard is given to plasma levels of corticosteroid-binding globulin (CBG). We have developed and validated an enzyme-linked immunosorbent assay (ELISA) for CBG and together with plasma cortisol calculated the "free cortisol index" as an additional parameter for assessing adrenal function. METHODS: A monoclonal antibody was raised against CBG. The antibody was characterized by Western blotting and used with a polyclonal antibody to develop a direct ELISA for CBG. Together with total plasma cortisol, the free cortisol index was derived and correlated with an "in-house" ligand binding method for assessing free cortisol. The free cortisol index was also used as an adduct to total plasma cortisol in assessing adrenal function. RESULTS: The ELISA has acceptable performance and the free cortisol index correlates well with free cortisol determined by ligand binding. In addition, we show that CBG levels following synthetic ACTH (synacthen) show a modest but significant decline. CONCLUSION: We conclude that the measurement of both plasma CBG and total cortisol to derive the free cortisol index may provide an additional parameter in the interpretation of the short synacthen test and that there is a decline in plasma CBG over this test. PMID- 12559608 TI - Establishment of age-dependent reference values for IgA subclasses. AB - BACKGROUND: Recently, subclass-specific antisera have been introduced for application in a nephelometric assay. The aim of this study was to establish age dependent reference values for serum concentrations of the two IgA subclasses in children and adults. METHODS: Serum levels of IgA1 and IgA2 were measured by automated immunonephelometry in samples from 235 clinically healthy children between 6 months and 18 years of age and 36 healthy adults. RESULTS: Both IgA1 and IgA2 were detectable in all samples, and both IgA1 and IgA2 increased with increasing age. In adults, the mean value for IgA1 is 1.46 g/l for IgA2 0.21 g/l and for total IgA 1.94 g/l. Individual IgA2 values correlate significantly (p < 0.0001) with IgA1 values (r(2) = 0.5433). In addition, there was a highly significant (p < 0.0001) correlation (r(2) = 0.9530) between the measured total IgA and the sum of the two IgA subclasses indicating that immunonephelometry using highly specific polyclonal antisera might be superior to other methods. CONCLUSIONS: These results and the availability of age-dependent reference values make it worthwhile to reassess the role of IgA subclasses in immunodeficiency and autoimmune diseases where conventional methods have led to conflicting results. PMID- 12559609 TI - Antibodies to centromere antigens measured by an automated enzyme immunoassay. AB - BACKGROUND: Anticentromere antibodies (ACA) are frequently observed in patients with Raynaud's phenomenon and in the CREST syndrome, a subclass of systemic sclerosis. Likewise, ACA are also found in other autoimmune and non-autoimmune diseases. The objective of the present study was to evaluate the clinical utility of the measurement of antibodies to the best characterized centromere antigen (CENP-B) protein by an enzyme-linked immunosorbent assay (ELISA) that uses human recombinant CENP-B antigen and compare it with indirect immunofluorescence assay (IFA) on HEp-2 cells. METHODS: We have analyzed 128 sera samples from patients with the following diseases: systemic lupus erythematosus (SLE, n = 53), mixed connective tissue disease (n = 1), primary Sjogren syndrome (n = 10), primary Raynaud's phenomenon (n = 10), primary systemic sclerosis (n = 7), polymyositis/dermatomyositis (n = 3), rheumatoid arthritis (n = 9), cutaneous lupus (n = 5), primary biliary cirrhosis (n = 9), chronic autoimmune hepatitis (n = 5) and ANA-positive non-autoimmune diseases (n = 16). RESULTS: The ELISA evaluated shows a good concordance with IFA, with the advantage of being an automatable quantitative technique. CONCLUSIONS: Measurement of anticentromere antibodies by this ELISA using human recombinant antigen is a useful alternative for the autoimmune laboratory checking for diseases associated with anticentromere antibodies. PMID- 12559610 TI - Intra-individual variations of fasting plasma lipids, apolipoproteins and postprandial lipemia in familial combined hyperlipidemia compared to controls. AB - BACKGROUND: The intra-individual variability of plasma lipids and apolipoproteins has not been studied systematically in familial combined hyperlipidemia (FCHL). METHODS: Intra-individual changes in fasting plasma lipids and apolipoproteins B and AI were determined in 18 untreated FCHL subjects and 16 unrelated, normolipidemic subjects. Participants were matched for gender, age and body mass index. The mean follow-up period of fasting plasma lipids was 48.91 +/- 35.46 (mean +/- S.D.) days. Postprandial lipemia was determined on 3 different days in 1 week in 90 healthy controls and 17 untreated FCHL subjects by the area under the diurnal capillary triglyceride curve (TGc-AUC). RESULTS: The coefficients of variation (CVs) for fasting plasma TG were similar between FCHL (23.2 +/- 10.2%) and controls (20.4 +/- 8.2%). The CVs for HDL-C, apo B and apo AI were the lowest of all fasting plasma measurements in both groups and there was no significant difference between FCHL (12.8 +/- 8.2%, 13.2 +/- 15.8% and 6.4 +/- 5.2%, respectively) and controls (11.4 +/- 4.3%, 11.3 +/- 10.6% and 7.8 +/- 4.6%, respectively). The CVs for postprandial lipemia were not different between FCHL (15.9 +/- 11.3%) and controls (15.1 +/- 11.0%), and were significantly lower than the CV of fasting capillary TG (TGc) in the same period (36.3 +/- 24.7% and 24.9 +/- 17.2%, respectively). CONCLUSIONS: Our study does not provide evidence for short-term major changes in fasting or postprandial lipemia or apolipoproteins in FCHL when systematically compared to healthy controls. PMID- 12559611 TI - Quantitative PCR to evaluate small amounts of BCL2 mRNA in human peripheral T cells: implication of equimolar target and competitor end products. AB - BACKGROUND: It is difficult to reliably and reproducibly quantitate small amounts of mRNA by conventional PCR. The method we describe here enabled us to more accurately quantitate a small amount of BCL2 mRNA in human peripheral T cells. METHODS: The sample was amplified in four tubes containing three-fold serial dilutions of competitor so that when the PCR products in the four tubes were totaled, the number of target and competitor components were approximately equal. An unknown concentration of target molecules should be assessed only within a narrow range, requiring that several reactions be performed for each sample. RESULTS AND CONCLUSION: With this method, conventional PCR machines can be used to perform quantitative PCR on very small amounts of BCL2, and would be especially useful for samples that contain substance(s) that affect PCR amplification efficiency. PMID- 12559612 TI - Changes of oxidative stress in various tissues by long-term administration of vitamin E in hypercholesterolemic rats. AB - BACKGROUND: Free radical-mediated oxidative stress has been implicated in the genesis and progression of atherosclerosis. METHODS: The lipid peroxides and antioxidant status of various tissues were investigated in hypercholesterolemic rats and the effect of vitamin E supplementation on defense systems. RESULTS: Cholesterol-feeding caused a significant increase in the lipid peroxide concentrations of plasma, erythrocytes, liver and brain. In addition, a significant decrease in glutathione (GSH) content, glutathione peroxidase (GSH Px) and glutathione transferase (GSH-ST) activities were found in erythrocytes and liver but superoxide dismutase (SOD) activity remained unchanged in these tissues in comparison to the control group. Vitamin E supplementation to hypercholesterolemic rats induced a significantly decrease in lipid peroxide concentrations and a significant increase in the GSH content, GSH-Px and GSH-ST activities in erythrocytes and liver. CONCLUSIONS: Long-term administration of vitamin E may play an important role in suppressing oxidative stress, and thus, may be useful for the prevention and/or early treatment of hypercholesterolemia. PMID- 12559613 TI - Determination of urinary myo-inositol concentration by an improved enzymatic cycling method using myo-inositol dehydrogenase from Flavobacterium sp. AB - BACKGROUND: To determine myo-inositol more accurately, we improved the enzymatic cycling method. METHODS: We screened myo-inositol dehydrogenase (MIDH; EC.1.1.1.18) from Flavobacterium sp., which was highly specific to myo-inositol. We measured urinary myo-inositol/creatinine ratio 2 h after 75-g oral glucose tolerance test (2 h MI) of 71 volunteers, and investigated the relationship between diabetes and urinary myo-inositol concentration. RESULTS: The calibration curve was linear (r = 1.00) up to 2000 micromol/l, and the detection limit was 10 micromol/l. Within-run and between-run CVs were 0.5-1.1% and 0.4-1.3%, respectively. The 2 h MI of impaired fasting glycemia (IFG; 65.1 +/- 46.6 mg/g Cr, P < 0.005), impaired glucose tolerance (IGT; 85.0 +/- 73.7 mg/g Cr, P < 0.001) and diabetes (163.4 +/- 73.7 mg/g Cr, P < 0.0001) increased significantly compared with that of normal glucose tolerance (NGT; 24.0 +/- 14.4 mg/g Cr). From receiver operating characteristic analyses on 2 h MI, with 50 mg/g Cr as a tentative cutoff value to detect diabetes, the sensitivity and specificity were 100% and 77%, respectively. With 40 mg/g Cr as a tentative cutoff value to detect NGT, the sensitivity and specificity were 74% and 85%, respectively. CONCLUSIONS: The myo-inositol measurement method demonstrated high specificity and yielded accurate results. The results of clinical trials suggested that 2 h MI could not only determine diabetes but also distinguish IFG and IGT from NGT. PMID- 12559614 TI - Polymorphism of angiotensin-converting enzyme gene and BMI in obese Korean women. AB - BACKGROUND: The renin-angiotensin system (RAS) plays a role in the pathogenesis of metabolic diseases. This system was recently found to be completely expressed in human adipose tissue. Especially angiotensin II, the active component of RAS, may affect adipogenesis and adipocyte metabolism. We examined whether obese and non-obese subjects differ from angiotensin-converting enzyme (ACE) genotype distribution, and whether the ACE genotypes affect the anthropometric parameters or the degrees of body mass index (BMI). METHODS: The study included 155 obese healthy women (BMI > or = 25 kg/m(2), range 25-54.7, age range 15-40 years), 82 non-obese women (BMI < 25 kg/m(2), range 15-40 years), and 613 random controls. Total fat mass and percent body fat (PBF) were determined by dual-energy X-ray absorptiometry (DEXA). Genomic DNA was extracted and used for polymerase chain reaction (PCR)-based genotyping of ACE. RESULTS: Age, percent body fat, waist-to hip ratio (WHR), body mass index, and cholesterol concentrations did not differ from ACE genotype. No differences were observed for allelic and genotype frequencies between obese women (BMI > or = 25) and 82 non-obese women or 613 random controls. In addition, no association of ACE polymorphism was observed with BMI for genotype in obese women. CONCLUSIONS: ACE polymorphism is not a significant factor for BMI and does not contribute to the odds of obesity in obese healthy women from Korea. PMID- 12559615 TI - Enzymatic assay for determination of bicarbonate ion in plasma using urea amidolyase. AB - BACKGROUND: One of the important buffering systems to maintain blood pH is carbonic acid-bicarbonate. Together with other clinical tests, the measurement of bicarbonate ion concentrations is widely used for the diagnosis of the acid-base balance. We developed a kinetic assay for measurement of bicarbonate ion in plasma using urea amidolyase (EC 3.5.1.45) from yeast species. We evaluated the analytical performance of the present enzymatic method and examined the relationship between bicarbonate ion concentrations by present method and with ABL 520 blood gas system. METHODS: Urea amidolyase catalyzes the reaction of bicarbonate ion with urea to rise to allophanate. We eliminated endogenous ammonium ion by the use of glutamate dehydrogenase (EC 1.4.1.4), and then monitored the production of ammonium ion in the presence of urea amidolyase, urea, ATP, potassium, and magnesium ions. Ammonium ion was produced proportional to the bicarbonate ion concentration and was determined by adding glutamate dehydrogenase to produce NADP(+) in the presence of 2-oxoglutarate and NADPH, and the change of absorbance at 340 nm was monitored. RESULTS: The within-assay and day-to-day assay coefficient variations (CVs) of the present method were 1.3-2.8% and 3.1-5.4%, respectively. The analytical recoveries were 90-110%. The presence of ascorbic acid, bilirubin, hemoglobin, lipemic material, hydrogen phosphate, dihydrogen phosphate, ammonium, or calcium ion did not affect this assay. The correlation coefficient between the values obtained by present method (y) and Radiometer ABL 520 blood gas system (x) was 0.983 (y = 1.029x-0.737 mmol/l, Sy/x = 0.764, n = 100), with a mean difference of 0.03 +/- 0.77 mmol/l [(values by reference method-that of present method) +/- S.D.] using the Bland-Altman technique. PMID- 12559616 TI - Urinary iodine percentile ranges in the United States. AB - BACKGROUND: The status of iodine nutrition of a population can be determined by measurement of urinary iodine concentrations since it is thought to indicate dietary iodine intake. Normally, these results are compared to population-based criteria, since there are no reference ranges for urinary iodine. OBJECTIVE: To determine the percentile ranges for urinary iodide (UI) concentrations in normal individuals in the United States. MATERIALS AND METHODS: The third National Health and Nutrition Examination Survey (NHANES III) (1988-1994) database of the civilian, non-institutionalized, iodine-sufficient US population was used. The 2.5th to 97.5th percentile ranges for urinary iodine and for urinary iodine per gram creatinine ratio (UI/Cr) (microg/g) were calculated for females and males, 6 89 years of age, each stratified by age groups. RESULTS AND CONCLUSIONS: We calculated the percentile ranges for urinary iodine. After exclusions of subjects with goiter or thyroid disease, the study sample included 21,530 subjects; 10,439 males and 11,091 females. For women of childbearing age (14-44 years), urinary iodine concentration 2.5th to 97.5th percentiles are 1.8-65 microg/dl or 36-539 microg/g creatinine. For pregnant women, the ranges are 4.2-55 microg/dl or 33 535 microg/g creatinine. PMID- 12559617 TI - BNP test utilization for CHF in community hospital practice. PMID- 12559618 TI - A marked decrease in the activity of biotinidase in the plasma of rats after single dose of carbon tetrachloride. PMID- 12559619 TI - Fibrates and C-reactive protein. PMID- 12559620 TI - Pathobiochemistry and clinical use of procalcitonin. PMID- 12559621 TI - Assessment of killer cell immunoglobulinlike receptor expression and corresponding HLA class I phenotypes demonstrates heterogenous KIR expression independent of anticipated HLA class I ligands. AB - Natural killer (NK) cell-mediated cytolysis is stimulated and downregulated through the interaction of distinct human leukocyte antigen (HLA) class I molecules on target cells with specific killer cell immunoglobulinlike receptors (KIRs) on NK cells. Killer cell immunoglobulinlike receptors are highly polymorphic and are clonally distributed on NK cell populations within individuals. However, the regulation of KIR expression by individual HLA class I phenotypes is not well understood. To examine a potential influence of the HLA class I phenotype on KIR expression patterns we studied the KIR expression in individuals that were subgrouped according to the major HLA-C encoded KIR epitopes (group C1 versus C2). In these individuals, NK cells were analyzed for KIR expression using flow cytometry and RNA-based expression analysis. Our results demonstrate that KIR genes are transmitted very heterogeneously with two main patterns of KIR genotypes as previously described; group A and group B (with 21 different genotypes). There are distinct populations exhibiting different densities of CD158a and/or CD158b positive NK cells that coexist in all individuals. A clear correlation between KIR expression and the currently known HLA class I ligands was not observed. In conclusion, the surface expression of KIRs in individuals with different HLA class I genotypes indicates that other non HLA class I encoded factors contribute to the shaping of the KIR repertoire. PMID- 12559622 TI - Expansion of specific alphabeta+ T-cell subsets in the myocardium of patients with myocarditis and idiopathic dilated cardiomyopathy associated with Coxsackievirus B infection. AB - Idiopathic dilated cardiomyopathy (IDC) is one of the major causes of death in humans and has been linked to Coxsackievirus B (CVB) infection. The aim of this study was to analyze phenotypes of heart-infiltrating immune cells in patients suffering from myocarditis and IDC associated with CVB infections. We found that the myocardium of these patients was infiltrated by CD4(+) and CD8(+) T lymphocytes as well as macrophages. Evidence of CVB3/4 infections was also found. In the majority of patients, the T-cell receptor repertoire (TCR) of the infiltrating lymphocytes was restricted, with a polyclonal expansion of the Vbeta7 gene family. We also found that human leukocyte antigen (HLA) class II alleles associated with susceptibility to type 1 diabetes (HLA-DR4 and HLA DQA1*04/05/06 alleles) were remarkably infrequent in IDC patients (p < 0.005), thus suggesting that they might confer protection against IDC. Finally, mRNA for interleukin-1beta, interferon-gamma, and tumor necrosis factor-alpha was detected in the cardiac specimens, although at a lower level compared with specimens from hearts without signs of viral infections. We conclude that CVB infection of the human myocardium is associated with a selective, yet polyclonal activation of different T-cell subsets in genetically susceptible individuals. This immune response may play a critical role in modulating disease progression after viral infections. PMID- 12559623 TI - Derivation of HLA-B*0702 transgenic mice: functional CTL repertoire and recognition of human B*0702-restricted CTL epitopes. AB - Transgenic mice expressing chimeric human leukocyte antigen (HLA)-B*0702 and murine H-2K(b) class I molecules were evaluated as a model system to study the immunogenicity of human cytotoxic T lymphocyte epitopes. Immunization of these mice with six known HLA-B*0702-restricted cytotoxic T lymphocyte epitopes emulsified in incomplete Freund's adjuvant induced significant immune responses specific for all six epitopes. A comparison of the immune responses between HLA B*0702/K(b) and HLA-A*0201/K(b) transgenic mice demonstrated that the HLA B*0702/K(b) mice possess a T-cell receptor repertoire capable of recognizing human B*0702 epitopes. However, the magnitude of B*0702-specific responses induced in B*0702/K(b) mice were approximately tenfold lower than A*0201-specific responses induced in HLA-A*0201/K(b) transgenic mice. A panel of 24 B*0702 motif bearing peptides was used to examine the relationship between immunogenicity and HLA-B*0702 binding capacity. All seven peptides with high binding affinities of 50% inhibitory concentration < or =50 NM (IC(50) 50 nM or less) were immunogenic. Similarly, 75% (9 of 12) of the intermediate binders (IC(50) nM of 50-500) were also immunogenic. Finally, only two of five peptides with binding capacity > 500 nM were found to have marginal immunogenicity, whereas the other three were completely negative. HLA-B*0702/K(b) transgenic mice were found to induce B*0702 specific responses after immunization with whole DNA genes or minigenes, suggesting that, at least to some degree, B*0702 epitopes were generated as a result of natural in vivo processing and presentation. PMID- 12559624 TI - Lack of optimal T-cell reactivity against the hepatitis C virus is associated with the development of fibrosis/cirrhosis during chronic hepatitis. AB - Chronic hepatitis C virus (HCV) infection develops in 85% of exposed individuals and 20% develop cirrhosis. However, the pathogenesis of this process is not well understood. The objective of this study was to determine whether HCV-reactive T cells play a role in the process of development of cirrhosis during chronic HCV infection. We analyzed 21 human leukocyte antigen (HLA)-A2 patients with chronic HCV infection (9 with histology of inflammation and 12 with histology of fibrosis/cirrhosis). The frequency of CD8(+) T cells reactive to 12 HCV-derived epitopes was determined by an interferon-gamma enzyme-linked immunospot (ELISPOT) assay. The frequency of CD4(+) Th1 and Th2 cells reactive to the HCV core antigen was determined by interferon-gamma and interleukin-5 ELISPOT assays, respectively. Patients with histology of inflammation showed a significantly higher CD8(+) T-cell response to five HCV-derived epitopes (YLLPRRGPRL [core], CINGVCWTV [NS3], LLCPAGHAV [NS3], ILAGYGAGV [NS4B], and GLQDCTMLV [NS5B]) as compared with patients with histology of fibrosis/cirrhosis. Also, patients with histology of inflammation showed a significantly higher CD4(+) Th1 response to the HCV core antigen as compared to patients with histology of fibrosis/cirrhosis. These results indicate that a lack of an optimal T-cell response to HCV is associated with the development of cirrhosis during chronic HCV infection. PMID- 12559625 TI - HCMV glycoprotein US6 mediated inhibition of TAP does not affect HLA-E dependent protection of K-562 cells from NK cell lysis. AB - Human cytomegalovirus has evolved multiple strategies to interfere with immune recognition by the host. A variety of mechanisms affect antigen presentation by major histocompatibility complex class I molecules resulting in a reduced class I cell-surface expression. This downregulation is expected to trigger natural killer (NK) cytotoxicity, requiring counteraction by the virus to establish long term infection. Here we describe that the human cytomegalovirus gpUS6 protein, which has been demonstrated to downregulate the expression of human leukocyte antigen (HLA) class I and the presentation of cytotoxic T lymphocyte epitopes by blocking transporter associated with antigen presentation (TAP function), does not affect the ability of HLA-E to inhibit NK cell mediated lysis of K-562 cells by interaction with CD94/NKG2A expressed on NK cells. Cell surface expression and function of HLA-E is not altered although gpUS6 inhibits TAP-dependent peptide transport by 95%. Moreover, HLA-E molecules presenting HLA class I signal sequence-derived peptides are functionally detectable on transfected TAP deficient RMA-S cells. PMID- 12559626 TI - Specific stimulation of MHC-transgenic mouse T-cell hybridomas with xenogeneic APC. AB - From the recombinant human leukocyte antigen (HLA)-DR1/H2-E(k) major histocompatibility complex (MHC) class II-transgenic mice, we have generated two CD4(+) T-cell hybridomas specific for peptides which were derived from human prostatic acid phosphatase (PAP) complexed to the human class II molecule HLA DR1. Both hybridomas strongly react to PAP-pulsed antigen-presenting cells (APC) from transgenic mice. Interestingly, these hybridomas also responded to PAP antigen presented by HLA-DR1-positive human APC. The species-mismatched T-cell stimulation occurs despite the biologic discordance in participating accessory molecules, which are required for the optimal T-cell-APC interaction. Our results demonstrate various degrees of functional interaction between coreceptors, costimulatory molecules, and integrins, which are expressed on the surface of T cell hybridomas and heterologous APC. PMID- 12559627 TI - Competition-based cellular peptide binding assays for 13 prevalent HLA class I alleles using fluorescein-labeled synthetic peptides. AB - We report the development, validation, and application of competition-based peptide binding assays for 13 prevalent human leukocyte antigen (HLA) class I alleles. The assays are based on peptide binding to HLA molecules on living cells carrying the particular allele. Competition for binding between the test peptide of interest and a fluorescein-labeled HLA class I binding peptide is used as read out. The use of cell membrane-bound HLA class I molecules circumvents the need for laborious biochemical purification of these molecules in soluble form. Previously, we have applied this principle for HLA-A2 and HLA-A3. We now describe the assays for HLA-A1, HLA-A11, HLA-A24, HLA-A68, HLA-B7, HLA-B8, HLA-B14, HLA B35, HLA-B60, HLA-B61, and HLA-B62. Together with HLA-A2 and HLA-A3, these alleles cover more than 95% of the Caucasian population. Several allele-specific parameters were determined for each assay. Using these assays, we identified novel HLA class I high-affinity binding peptides from HIVpol, p53, PRAME, and minor histocompatibility antigen HA-1. Thus these convenient and accurate peptide binding assays will be useful for the identification of putative cytotoxic T lymphocyte epitopes presented on a diverse array of HLA class I molecules. PMID- 12559629 TI - Antibodies to anionic phospholipids and anti-beta2-GPI: association with thrombosis and thrombocytopenia in systemic lupus erythematosus. AB - In this study, we evaluated the prevalence and association with thrombosis and/or thrombocytopenia of IgG and IgM antibodies to cardiolipin (aCL), phosphatidic acid (aPA), phosphatidylinositol (aPI), phosphatidylserine, and beta(2) glycoprotein I (abeta(2)-GPI) in systemic lupus erythematosus (SLE). Sera were obtained from 87 patients affected by SLE (77 of the 87 patients were females), 41 of them with a history of arterial and/or venous thrombosis. Antiphospholipid antibodies and abeta(2)-GPI were evaluated by enzyme-linked immunosorbent assay. IgG-aCL, IgG-aPA, IgG-aPI, IgG-aPS, and IgG-abeta(2)-GPI were found in 53%, 37%, 32%, 38%, and 24% of patients, respectively. IgM-aCL, IgM-aPA, IgM-aPI, IgM-aPS, and IgM-abeta(2)-GPI were detected in 15%, 17%, 18%, 14%, and 16%, respectively. With respect to antibody titer, IgG-aCL strongly correlated with all other antiphospholipid antibodies and abeta(2)-GPI of IgG isotype. Thrombosis was significantly associated with IgG-aPA (p = 0.044), IgG-aPI (p = 0.038), IgG-aPS (p = 0.026), IgG-abeta(2)-GPI, IgM-aPA (p = 0.044), IgM-aPI (p = 0.024), and IgM aPS (p = 0.01), irrespective of antibody titer, whereas IgG-aCL were associated with thrombosis and thrombocytopenia when taken at medium-high titer (p = 0.009 and p = 0.046, respectively). Our results confirm that, besides aCL and abeta(2) GPI, other antibodies to negatively-charged phospholipids are present in a large percentage of patients with SLE. However, it remains doubtful whether these other antiphospolipid antibodies actually represent an important parameter predictive of thrombosis and thrombocytopenia in SLE. PMID- 12559628 TI - Mild acid treatment induces cross-reactivity of 4H84 monoclonal antibody specific to nonclassical HLA-G antigen with classical HLA class I molecules. AB - Mild acid treatment by releasing beta(2)m and antigenic peptides leaves human leukocyte antigen (HLA) class I free heavy chains attached to the cell surface. Acid treatment thus allows detection of the cell surface class I antigens by monoclonal antibodies (mAbs) specific to HLA-free heavy chains. We found that acid treatment also enables detection of the cell surface non-classical HLA-G class I antigen with mAbs specific for HLA-G free heavy chains, including 4H84 mAb recognizing all isoforms. Furthermore, we found that 4H84 mAb, but not other mAbs specific to HLA-G free heavy chains, binds to the surface of 8 out of 16 acid-treated leukemia cell lines. Nevertheless, HLA-G antigen is not present in any of these leukemia cells. This was demonstrated by failure to detect any antigen with 4H84 mAb in immunoblotting as well as by inability to detect HLA-G mRNA by RT-PCR. The antigen recognized by 4H84 mAb in some acid treated leukemia cells was identified by immunoprecipitation as a 45 kDa protein. A number of observations indicate that 45 kDa proteins are none other than classical class I heavy chains. Acid treatment thus induces the ability of the 4H84 mAb to recognize some classical HLA class I molecules. Remarkably, 4H84 determinant on HLA-G is linear but corresponding determinant present on some partially folded classical HLA class I free heavy chains is conformational. In view of the unexpected cross-reactivity, detection of HLA-G with this mAb must be carefully evaluated to avoid false detection. PMID- 12559631 TI - Fas ligand gene polymorphisms are not associated with Hashimoto's thyroiditis and Graves' disease. AB - Hashimoto's thyroiditis and Graves' disease represent the two most common autoimmune thyroid disorders. Whereas in Hashimoto's thyroiditis FasL expression causes thyrocytes to undergo apoptosis, additional anti-apoptotic molecules appear to protect these cells in Graves' disease. Mutations of the FasL gene were observed in systemic lupus erythematosus. Given its functional relevance for the pathogenesis of thyroid autoimmunity we wondered whether variants of the FasL gene play a role in Hashimoto's thyroiditis and Graves' disease. We genotyped families with at least one offspring affected by Hashimoto's thyroiditis (n = 86) and Graves' disease (n = 90) for two FasL gene polymorphisms (C -843 T in the promoter, A IVS2nt-124 G in intron 2). Extended transmission disequilibrium (ETDT) and chi(2) testing were performed. Neither polymorphism alone nor the promoter/intron 2 haplotypes (p = 0.91) were associated with Hashimoto's thyroiditis. No association with Graves' disease was observed for the promoter polymorphism (p = 0.91) and the intron 2 "A" allele (57.1%; p = 0.36) or the promoter/intron 2 haplotypes (p = 0.31). Moreover, intron 2 genotyping revealed no difference between an additional 251 patients with Graves' disease and 197 healthy controls (p = 0.37). Italian and German families did not differ for the studied polymorphisms. In conclusion, our data do not suggest common genetic FasL variants to significantly contribute to the pathogenesis of either Hashimoto's thyroiditis or Graves' disease. PMID- 12559630 TI - Prolactin and prolactin receptor gene polymorphisms in multiple sclerosis and systemic lupus erythematosus. AB - Genes encoding for prolactin (PRL) and its receptor (PRLR) are possible candidates for multiple sclerosis (MS) and systemic lupus erythematosus (SLE) susceptibility. In fact: (1) a prolactin secretion dysfunction has been described in several autoimmune diseases including SLE and MS and their animal models; (2) both PRL and PRLR are structurally related to members of the cytokine/hematopoietin family and have a role in the regulation of the immune response; and (3) both PRL and PRLR genes map in genomic regions that showed linkage with autoimmunity. Prolactin maps on chromosome 6p, about 11-kb telomeric to HLA-DRB1 and PRLR in 5p12-13, which revealed evidence of linkage with MS in different populations. To evaluate a possible role of these two genes in SLE and MS we performed an association study of 19 PRL and PRLR single nucleotide polymorphisms (SNPs). These were directly searched by DHPLC in a panel of SLE and MS patients and selected from databases and the literature. The SNP allele frequencies were determined on patient and control DNA pools by primer-extension genotyping and HPLC analysis. Moreover a panel of HLA typed SLE and control individuals were individually genotyped for the PRL G-1149T polymorphism previously described to be associated with SLE. No statistically significant difference in the allele distribution was observed for any of the tested variations. PMID- 12559632 TI - MICA4/HLA-DRB1*04/TNF1 haplotype is associated with mixed connective tissue disease in Swedish patients. AB - In order to investigate major histocompatibility complex (MHC) class I chain related gene A (MICA), tumor necrosis factor (TNFa), -308TNFA, and human leukocyte antigen (HLA-DR/DQ) polymorphisms in mixed connective tissue disease (MCTD), we analyzed 24 patients and 229 healthy controls from Sweden. MICA and TNFa typing was performed by polymerase chain reaction (PCR) and genotyping. HLA DR and -DQ were genotyped using PCR-sequence specific primers (PCR-SSP) and PCR sequence-specific oligonucleotide probe (PCR-SSOP), respectively. For analysis of -308TNFA polymorphisms we performed PCR with restriction endonuclease enzymes. We found that the MICA5.1-5.1 genotype was positively associated with MCTD. Shared epitope genes (DRB1*01 and DRB1*04) were also significantly positively associated with MCTD. Polymorphism of -308TNFA was not differently distributed in MCTD patients compared with controls. Furthermore, we demonstrated that frequencies of three estimated haplotypes were increased in MCTD patients compared with controls. Interestingly, the haplotype with MICA allele 4 together with DRB1*04 and TNF1 alleles gives the most specific pattern for MCTD patients compared with controls. Our study demonstrates a clear contribution of HLA loci in susceptibility to MCTD in the Swedish population. Susceptibility to MCTD may be linked to the MICA4/HLA-DRB1*04/TNF1 haplotype and MICA 5.1-5.1 genotype. Mixed connective tissue disease was also associated with shared epitope genes, which in RA has been associated with a more severe disease. Whether these genotypes affect the clinical phenotype of MCTD needs to be determined. PMID- 12559633 TI - CTLA-4 +49 A/G dimorphism in Italian patients with celiac disease. AB - The chromosome region 2q33, which contains the cytotoxic T lymphocyte antigen-4 (CTLA-4) gene, has been reported in linkage and association with celiac disease (CD). In the present work we have tested the association between the polymorphism of the CTLA-4 exon 1 and susceptibility to CD in an Italian population, using case-control and family-based approaches. The +49 A/G dimorphism was analyzed in 86 patients, 144 ethnically matched controls, and 113 nuclear families by the polymerase chain reaction-restriction fragment length polymorphism method. A significantly higher frequency of the CTLA-4 +49A allele was observed in patients when compared with controls (p = 3 x 10(-2)). The segregation analysis in the 113 trios showed a preferential transmission of the A allele to the probands (chi(2)(TDT) = 4.85). When the patients were stratified according to the presence/absence of the high-risk human leukocyte antigen-DQ2 heterodimer, a significant difference was observed between the two groups, that is, the A allele was increased in the subjects without the DQ2 heterodimer (88.9% vs 73.5%, p = 8.3 x 10(-3)). The A allele was transmitted from heterozygous parents to eight of nine DQ2-dimer-negative patients. These data support CTLA-4 as a predisposing gene for CD in an Italian population with a prominent role in patients not carrying the high-risk human leukocyte antigen-DQ2 molecules. PMID- 12559635 TI - A new HLA-DRB allele (DRB1*15014) identified in a Caucasian individual. AB - Polymerase chain reaction with sequence specific primers subtyping of a DRB1*15 allele in a male 11-year-old German Caucasian and his father revealed a reaction pattern that did not agree with any known specificity, thus suggesting the existence of a novel allele. Sequence-based typing after allelic separation revealed the new allele human leukocyte antigen (HLA)-DRB1*15014. The sequence pattern of HLA-DRB1*15014 might have been generated as a result of two independent recombinations, most likely over several generations. PMID- 12559634 TI - TNF, TNF receptor type 1, and allograft inflammatory factor-1 gene polymorphisms in Japanese patients with type 1 diabetes. AB - The human leukocyte antigen (HLA) class III region, located on chromosome 6p21, has been regarded as one of the susceptible loci for type 1 diabetes. Because it contains many genes related to inflammatory and immune responses, including tumor necrosis factor (TNF), lymphotoxin-alpha (LT-alpha), and allograft inflammatory factor 1 (AIF-1) genes, it is unclear which gene within the class III region is responsible for the susceptibility to the disease. We sequenced the AIF-1 gene region and detected three novel polymorphisms, all of which were diallelic and localized at introns. Then, we investigated AIF-1, TNF, and LT-alpha gene polymorphisms in 165 patients with type 1 diabetes, consisting of 90 patients with young-onset type 1 diabetes, 75 patients with adult-onset type 1 diabetes, and 200 control patients. We also analyzed TNF receptors type 1 (TNFR1) and type 2 (TNFR2) gene polymorphisms, located on chromosome 12p13 and 1p36, respectively. Although there were significant differences between type 1 diabetes patients and controls in the distributions of TNF promoter polymorphisms at position -1031 and -857, and LT-alpha gene NcoI polymorphism, none of them was independently associated with the disease after two-locus analysis with HLA class II alleles. We detected the significantly increased frequency of the -383C allele, located in the TNFR-1 promoter region, in both young-onset and adult-onset diabetes patients compared with controls. In addition, the -383C allele was found to be associated with higher expression of the TNFR1 gene than that of -383A allele in in vitro expression. These results suggest that the TNFR1 gene region might be a susceptible locus to type 1 diabetes in Japanese. PMID- 12559636 TI - Nomenclature for factors of the HLA system, update October 2002. PMID- 12559638 TI - Practical midcourse sample size modification in clinical trials. AB - Power calculations are very important in the planning of a well-designed clinical trial. Sometimes there is limited information available before the trial, making it highly desirable to adjust the sample size after seeing actual trial data. Indeed, there has been a recent proliferation of papers promising great flexibility in midcourse correction of sample size and other design features, such as choice of primary endpoint. We point out the difficulty in accurately estimating the treatment effect midway through a trial, and we encourage the use of a simple, conservative approach whereby sample size can be increased but not decreased from what was originally planned. We show how to compute the p value and confidence interval for this two-stage procedure. If the original sample size is maintained, analysis of the data is the same as for a fixed sample procedure. PMID- 12559639 TI - Monitoring mortality at interim analyses while testing a composite endpoint at the final analysis. AB - Mortality is often used as the clinical endpoint in clinical trials for acute diseases and takes precedence over any other outcome. A composite outcome such as death plus disease occurrence (or recurrence) or death plus hospitalization may also be considered, sometimes even as the primary outcome due to practical sample size issues. That is, a composite endpoint should have a higher event rate and thus a smaller sample size than for mortality alone to reach the same power. Two different scenarios are considered: in Scenario 1, the composite outcome is the primary endpoint and the mortality outcome is secondary; in Scenario 2, the mortality outcome is the primary endpoint and the composite outcome is secondary. In either scenario, the trial will be stopped if the simple mortality outcome shows an adverse effect or a significant benefit at an interim analysis, while the composite outcome will be tested at the final analysis if the mortality outcomes fails to show significance. These scenarios are typical in many trials sponsored by industry for regulatory approval. We refer to them as a switching the primary endpoint process. Two switching-endpoint procedures are proposed to calculate the efficacy boundary for the composite test statistic at the final analysis. The Bonferroni method is used in Method 1. In Method 2, the calculation is based upon the joint distribution of the test statistics for the simple mortality and the composite outcomes. A completed clinical trial, prospective randomized amlodipine survival evaluation (PRAISE-1), is used to illustrate the two switching-endpoint procedures. A simulation study shows that the two switching-endpoint procedures allow a trial to be stopped early due to a clinically relevant benefit in the mortality while preserving the overall alpha level. PMID- 12559640 TI - Statistical issues related to early closure of STOP-ROP, a group-sequential trial. AB - The Supplemental Therapeutic Oxygen for Prethreshold Retinopathy of Prematurity trial used the group-sequential alpha-spending approach with asymmetrical tails to investigate whether supplemental oxygen therapy would reduce the proportion of infants progressing from prethreshold retinopathy of prematurity (ROP) to threshold ROP from 30% to 20%. Three years of enrollment were predicted. After 4 years, the data and safety monitoring committee (DSMC), faced with a projected delay of 2-3 more years, opted to terminate the trial after 1 further year of enrollment despite a continually borderline test statistic. We discuss factors relating to the DSMC decision and its effect upon the conclusions that may be drawn from the trial. PMID- 12559641 TI - Stopping the active intervention: CARET. AB - The Carotene and Retinol Efficacy Trial (CARET) was a large, multicenter randomized chemoprevention trial designed to test a combined lung cancer prevention agent in heavy smokers and workers exposed to asbestos. In January 1996, the CARET Steering Committee decided to stop the intervention due to an adverse effect. This paper describes the decision process used to apply the stopping rules and the activities engaged in by CARET participants and staff to implement the decision. The most important activity was to draft and mail a letter to the participants informing them of the disappointing CARET results and asking them to stop taking the study vitamins and to return any unused study vitamins. The steering committee, with the support of the National Cancer Institute, planned to follow participants for disease endpoints and smoking behavior for 5 years. These activities led to smooth closure of active intervention and maintained high retention rates during the transition. PMID- 12559642 TI - A model for the interim analysis process: a case study. AB - To evaluate data from a clinical trial before its completion, researchers routinely perform interim analyses. However, if not performed carefully, interim analyses can compromise the integrity of a clinical trial. In the last 10-15 years, regulatory authorities and the pharmaceutical industry have developed procedures and guidelines to allow trial sponsors access to unblinded data in an ongoing clinical trial without affecting the outcome. In December 1996, Abbott Laboratories was codeveloping a drug for treatment of an autoimmune disease. The pivotal phase II/III trial for the new drug application was very expensive, large, long term, and slow-accruing. The trial was initiated with a great deal of uncertainty concerning the safety and efficacy of the proposed treatment. An interim analysis was a logical part of the trial design. Two interim analyses were performed; the second analysis resulted in early termination of the trial. This article describes the interim analysis, including the process used for planning and execution and the lessons learned from the experience. In addition, the methodology for performing an interim analysis and the roles and responsibilities of involved members are discussed. PMID- 12559643 TI - Be skeptical about unexpected large apparent treatment effects: the case of an MRC AML12 randomization. AB - The preliminary results of the twelfth Medical Research Council acute myeloid leukemia trial show no evidence of a survival advantage for five courses of therapy compared to four courses in a randomized comparison involving 1078 patients (hazard ratio 1.09, 95% confidence interval [CI] 0.87-1.37, p=0.4). However, the data presented to the independent data monitoring and ethics committee (DMEC) at both its reviews in 1998 suggested large benefits for the additional course with hazard ratios of 0.47 and 0.55 (95% CIs 0.29-0.77 and 0.38 0.80, p=0.003 and p=0.002, respectively). Despite these highly significant findings, the DMEC did not recommend closure of the randomization, a decision vindicated by the subsequent reversion to a null result. The main reason for not closing the randomization was that the treatment effects observed in 1998 (53% and 45% reductions in the odds of death) were considered too large to be clinically plausible, despite the p-values associated with them. Investigations have not identified any clinical explanations, such as different types of patients in the early and later parts of the trial, to explain the loss of benefit as the trial progressed. Thus, the most likely current explanation for the large benefit observed early on is the play of chance. Lessons to be learned from this example are that: fixed stopping rules based on some predetermined p value should not be used and the decision to close a randomization or not should take account of other factors such as the medical plausibility of the magnitude of the treatment effect; chance effects do occur and happen more frequently than many clinicians realize; it is important that DMEC members are experienced in the interpretation of clinical trial evidence and aware of the dangers of early stopping without wholly convincing evidence. PMID- 12559644 TI - Data and safety monitoring board issues raised in the VA Status Epilepticus Study. AB - The Department of Veteran Affairs Status Epilepticus Cooperative Study was a randomized, multicenter clinical trial testing four intravenous drug regimens (lorazepam, phenobarbital, phenytoin and diazepam followed by phenytoin) to treat generalized convulsive status epilepticus. During the course of the study, two problems emerged that the study's data and safety monitoring board (DSMB) was required to address: poor recruitment and an unexpected difference in 30-day mortality between treatment groups. By the first annual DSMB meeting, recruitment was only 25.6% of expected. The DSMB recommended placing the study on probation and replacing poorly performing sites. At their second annual meeting, the DSMB recommended approval of proposed changes to the study design contingent on the study leadership removing nonproductive sites. These changes were a 2-year increase in the recruitment period and a change in study design that decreased required sample size. Nonproductive centers were terminated and the approved changes allowed the study to be successfully completed. At the second annual DSMB meeting, an unexpected doubling of mortality rates between drug groups was observed. Although not statistically significant, the finding raised serious concerns for patient safety. The DSMB recommended instituting monthly reporting on mortality and suggested additional analyses for exploring why the differences could be occurring. These analyses indicated that, by chance, older and sicker patients were being randomized to the drugs with the higher mortality rates. By the end of the study, the observed differences in mortality between drug groups had evened out. The DSMB's thoughtful recommendations, support and monitoring ensured that the study was successfully completed without endangering the study patients. PMID- 12559645 TI - Termination of a randomized controlled trial of two vasectomy techniques. AB - Family Health International (FHI) and EngenderHealth conducted a multicenter, randomized controlled trial to evaluate a fascial interposition (FI) component of a vas occlusion procedure for male sterilization. A data monitoring committee (DMC) was established to provide independent review of the interim report. The DMC met to review the interim report after enrollment of 552 men. As planned, an estimate of the hazard ratio (HR) for successful vasectomy for the FI group versus the no-FI group controlling for age of the participant and level of experience of the surgeon was obtained for the interim report using Cox's proportional hazards regression. The main analysis comparing time to vasectomy success, defined as two consecutive azoospermic semen samples, was highly significant and met the prespecified level for stopping enrollment (HR=1.54, p<0.01). However, a troublesome age by vasectomy technique interaction became apparent in the analysis. FI was clearly beneficial for younger males. However, the trend for older men was not as clear. The DMC recommended continuing the trial to better study this interaction and obtain more data to clarify the effect of FI for older men. After consulting further with the DMC, the study statisticians presented the interim report to senior management at FHI. A meeting between FHI senior management and the DMC followed. After much deliberation, trial enrollment was terminated. Follow-up of participants already enrolled in the study continued as planned. This paper presents the issues involved in the decision to terminate the study. PMID- 12559646 TI - DSMB case study: decision making when a similar clinical trial is stopped early. AB - Two similarly designed, placebo-controlled clinical trials are fully accrued and following patients for safety and outcome data. One trial is stopped at a planned interim analysis by its data safety and monitoring board (DSMB) due to a statistically significant treatment benefit. The statisticians and DSMB of the other trial are informed of these results. What are the responsibilities of the statistical center? How should the DSMB deal with the situation if the data do not support the stopped trial? What should the patients be told concerning the results of the two trials when one trial continues and the other is stopped? This DSMB case study reports on such a situation for two randomized clinical trials of oral ganciclovir for the prevention of cytomegalovirus disease in HIV/AIDS patients. PMID- 12559647 TI - Data and safety monitoring board deliberations resulting in the early termination of the Monoclonal Antibody Cytomegalovirus Retinitis Trial. AB - We report on two features of the early termination of the studies of ocular complications of AIDS monoclonal antibody cytomegalovirus retinitis trial: one related to the deliberative process of the treatment effects monitoring body (the policy and data monitoring board [PDMB]) and the other related to the relationships among the PDMB, the investigators, and the joint sponsors (the National Institutes of Health and a pharmaceutical company). The PDMB was challenged with reconciling internally inconsistent safety and mortality data and determining the weight to give mortality data from other concurrently running trials. The coordinating center faced a challenge in negotiating how to communicate results from a jointly sponsored trial. Early resolution of certain organizational and procedural issues, such as rules regarding absentee voting, to which body the monitoring committee should report officially, and the timing and general content of dissemination of different kinds of results, would make jointly sponsored trials more robust to difficulties at the final hour. PMID- 12559651 TI - Is there a role for 5-HT1A agonists in the treatment of depression? AB - The role of serotonin (5-hydroxytryptamine; 5-HT) in the treatment of depressive and anxiety disorders is underscored by the therapeutic action of selective 5-HT reuptake inhibitors acting to enhance the degree of activation of various 5-HT receptor subtypes. The 5-HT1A receptors are particularly relevant to the antidepressant and anxiolytic responses in human beings. They are located presynaptically in the raphe nuclei, where they act as cell body autoreceptors to inhibit the firing rate of 5-HT neurons, and are located postsynaptically in limbic and cortical regions, where they also attenuate firing activity. The azapirones are full agonists at 5-HT1A autoreceptors and are generally, but not exclusively, partial agonists at postsynaptic 5-HT1A receptors. Some of these drugs, including gepirone and other 5-HT1A agonists such as buspirone, have been reported to exert anxiolytic and antidepressive activity in double-blind, placebo controlled, and comparative trials. Their delayed therapeutic activity is believed to result from increased activation of postsynaptic 5-HT1A receptors occurring only after 5-HT neurons regain their normal firing activity. The recovery of this parameter, which is attributable to 5-HT1A autoreceptor desensitization, also restores 5-HT release. At this point, the summed effects of a normalized level of synaptic 5-HT and the exogenous 5-HT1A agonist can be exerted on postsynaptic 5-HT1A receptors. The widespread recognition of the clinical efficacy of such agents has largely been hampered by their undesirable pharmacokinetic properties. Most 5-HT1A agonists are indeed readily absorbed but are also rapidly eliminated, thereby often producing either suboptimal therapeutic responses at low doses, or cumbersome adverse effects at higher doses. Extended-release formulations allow once-daily dosing regimens, thus avoiding sharp peak plasma concentrations. This improves compliance and permits the use of higher dosages, which may be associated with enhanced efficacy and better tolerability relative to the immediate-release formulations. In sum, 5 HT1A receptor agonism represents a valuable and efficacious therapeutic approach to major depression. PMID- 12559648 TI - Unique roles of a data and safety monitoring board in vaccine safety trials with compressed timelines and urgent implications. AB - Studies with urgent implications for public policy and programs represent a special case for monitoring boards, whose responsibilities must include ensuring applicability to policy making and timely communication of results as well as data and safety monitoring. We recently conducted two preseason evaluations of an influenza vaccine, results of which would influence the supply of vaccine for the Canadian annual program. Only 12 weeks were available for planning the studies and 6 weeks for fieldwork and data analysis. The enlisted board played key roles in determining appropriate study design, setting and enforcing rules for study initiation and early termination, monitoring safety, and encouraging timely dissemination of results. An extended role in data monitoring would have been desirable. Providing a safety valve for inevitable tensions between investigators and sponsor was an understated but significant role. Lessons from our experience will be particularly pertinent to future studies of pandemic influenza vaccines. PMID- 12559652 TI - Recall of emotional states in posttraumatic stress disorder: an fMRI investigation. AB - BACKGROUND: The goal of this study was to examine the neuronal circuitry underlying different emotional states (neutral, sad, anxious, and traumatic) in posttraumatic stress disorder (PTSD) in traumatized subjects versus traumatized subjects without PTSD. METHODS: Traumatized subjects with (n = 10) and without (n = 10) PTSD were studied using the script-driven symptom provocation paradigm adapted to functional magnetic resonance imaging (fMRI) at a 4 Tesla field strength. RESULTS: Compared to the trauma-exposed comparison group, PTSD subjects showed significantly less activation of the thalamus and the anterior cingulate gyrus (area 32) in all three emotional states (sad, anxious, and traumatic). CONCLUSION: These findings suggest thalamic and anterior cingulate dysfunction in the recollection of traumatic as well as other negative events. Thalamic and anterior cingulate dysfunction may underlie emotion dysregulation often observed clinically in PTSD. PMID- 12559653 TI - Activation of the medial prefrontal cortex and extended amygdala by individual ratings of emotional arousal: a fMRI study. AB - BACKGROUND: Significant differences between individual responses to emotional stimuli can be important for the study of emotion. We investigated whether incorporating individual ratings of emotional arousal in the analysis of functional magnetic resonance imaging (fMRI) data improves the detection of activation in the medial prefrontal cortex (MPFC) and sublenticular extended amygdala (SLEA), areas implicated in the processing of emotional salience. METHODS: Healthy subjects viewed counterbalanced blocks of aversive, nonaversive, and blank images. Outside the scanner, they rated the intensity of emotional arousal (salience) of each presented picture. RESULTS: Incorporating the subject's response to each stimulus by using individualized regressors produced more robust activations within MPFC and SLEA compared with a simple boxcar regressor, identical for all subjects. CONCLUSIONS: Our findings demonstrate that individual behavioral data are useful in improving detection of activation in block-design functional imaging studies. PMID- 12559654 TI - Temporal instability of auditory and visual event-related potentials in posttraumatic stress disorder. AB - BACKGROUND: We examined P300 measures in patients with posttraumatic stress disorder (PTSD) and control subjects at two different time points to determine event-related potential (ERP) stability over time and the relationship of changes in ERPs to changes in symptom levels. METHODS: Auditory and visual P300 was recorded in a three-condition novelty oddball task in 25 male subjects with combat-related PTSD and 15 male combat-exposed normal control subjects at two time points separated by 6-12 months. Regression analyses were conducted to compare the temporal stability of ERP measures in PTSD and control subjects. Variability in ERP measures over time within PTSD subjects was examined for association with changes in symptom levels. RESULTS: There were no significant differences in P300 amplitude or latency in PTSD versus control subjects at either time point, regardless of stimulus type (target, novel) or modality (auditory, visual). Nine of 24 P300 measures were significantly less predictable over time in the PTSD group compared to control subjects. Variability of P300 measures over time was not associated with fluctuations in symptoms of depression or PTSD. CONCLUSIONS: P300 ERPs are more variable cross-sectionally and over time in PTSD subjects compared to trauma exposed control subjects. Measures of variability about the group mean appear to be more informative about the cognitive electrophysiology of PTSD than measures of central tendency. PMID- 12559655 TI - Task instructions modulate neural responses to fearful facial expressions. AB - BACKGROUND: The amygdala, hippocampus, ventral, and dorsal prefrontal cortices have been demonstrated to be involved in the response to fearful facial expressions. Little is known, however, about the effect of task instructions upon the intensity of responses within these regions to fear-inducing stimuli. METHODS: Using functional magnetic resonance imaging, we examined neural responses to alternating, 30-sec blocks of fearful and neutral expressions in nine right-handed male volunteers during three different 5-min conditions: 1) passive viewing; 2) performance of a gender-decision task, with no explicit judgment of facial emotion; 3) performance of an emotionality judgment task - an explicitly emotional task. RESULTS: There was a significant effect of task upon activation within the left hippocampus and the left inferior occipital gyrus, and upon the magnitude of response within the left hippocampus, with maximal activation in these regions occurring during passive viewing, and minimal during performance of the explicit task. Performance of the gender-decision and explicit tasks, but not passive viewing, was also associated with activation within ventral frontal cortex. CONCLUSIONS: Neural responses to fearful facial expressions are modulated by task instructions. PMID- 12559656 TI - Improvement in learning associated with increase in hippocampal formation volume. AB - BACKGROUND: Patients with spontaneous Cushing's syndrome are exposed to elevated levels of endogenous cortisol for months to years. We previously reported that hippocampal formation volume (HFV) increased in such patients after treatment lowered cortisol to normal concentrations. In the present study, we examined whether the structural increase was associated with improvement in cognition. METHODS: Twenty-four patients with Cushing's disease were studied before treatment and following treatment. Magnetic resonance imaging was used to measure HFV and caudate head volume. Neuropsychologic tests of verbal cognition, learning, and memory were also administered. RESULTS: Patients showed variability in improvement on neuropsychologic test performance. After partialing out age, education, duration of illness, and time since surgical treatment, greater improvement in word list learning, as measured by the Selective Reminding Test was associated with greater increase in HFV (r =.59, p <.02). There were no significant associations between improvement in paragraph or paired-word learning or memory tasks and increase in HFV. Improvement in other verbal tasks not strongly dependent on the hippocampus were not significantly associated with increase in HFV. CONCLUSIONS: After cortisol levels decline to normal concentrations, structural volumetric increase in HFV is accompanied by functional improvement in learning of unrelated words. PMID- 12559657 TI - Additional, physically ordered markers increase linkage signal for bipolar disorder on chromosome 18q22. AB - BACKGROUND: We recently reported evidence of linkage of bipolar disorder to chromosome 18q, with a paternal logarithm of odds (LOD) score of 4.67 (p =.004) in a clinically defined subset of families. Like other linkage studies, we had to rely on imprecise genetic maps to establish the marker order. Here, we test for linkage in the same sample with a denser set of markers, now physically ordered according to the draft sequence of the human genome. METHODS: Families were ascertained through probands with bipolar I disorder and diagnosed with reliable methods. Genotypes were generated for 12 microsatellite markers within an 11 centimorgan (cM) region of chromosome 18q22. Multipoint affected sib-pair linkage analysis was performed in a set of 16 nuclear families. RESULTS: The additional markers significantly increased the total genetic information extracted from our sample. We also observed an increase in the LOD score (to 5.42, p =.0066) and linkage resolution. The approximate 1-LOD support interval is now 9 male cM. CONCLUSIONS: The results strengthen our previous findings and further define a region suitable for genetic fine-mapping analysis on chromosome 18q. Our data suggest that a dense set of markers, when physically ordered, can increase the informational value of genetic linkage signals. PMID- 12559658 TI - Auditory sensory gating in hippocampus and reticular thalamic neurons in anesthetized rats. AB - BACKGROUND: Auditory gating is thought to reflect sensory information processing and is absent or diminished in schizophrenic patients. Although abnormal thalamic sensory processing has been proposed in schizophrenia, sensory gating of thalamic neurons has not been demonstrated experimentally. The aim of the present study was to establish whether auditory gating is present in the rat thalamus using a well-characterized animal model of auditory gating and schizophrenia. METHODS: Hippocampal electroencephalogram and single-unit activity in the thalamic reticular nucleus (nRT) were recorded in anaesthetized rats. Evoked potentials in the hippocampus and neuronal activity in the nRT were monitored in response to bilateral auditory stimuli. The effects of the psychostimulant D-amphetamine and the antipsychotic haloperidol on auditory gating were evaluated. RESULTS: Thalamic reticular nucleus neurons showed gated responses to paired-tone auditory stimuli, resembling hippocampal auditory gating. D-amphetamine disrupted auditory gating of nRT neurons and abolished their burst activity. D-amphetamine also disrupted hippocampal auditory gating and induced hippocampal theta activity. The amphetamine-induced gating deficit was reversed by haloperidol in both regions. CONCLUSIONS: Our findings provide the first experimental evidence for auditory gating of nRT neurons. We demonstrated that amphetamine disrupts sensory processing of nRT neurons, indicating similarities between hippocampal and thalamic sensory gating. These findings support the presumed connection between dopamine hyperfunction and abnormal thalamic filtering in schizophrenia. PMID- 12559659 TI - Alzheimer's disease and total plasma aminothiols. AB - BACKGROUND: Plasma homocysteine is elevated in Alzheimer's disease, but little is known regarding levels of related aminothiols in the disease. We therefore determined total plasma homocysteine, cysteine, and glutathione levels in patients and control subjects and investigated their relationship with cognitive scores. METHODS: We performed a prospective, case-controlled survey based in two UK Psychogeriatric Assessment Centres. Fifty patients with features compatible with DSM-IV criteria for primary degenerative dementia of Alzheimer type were recruited together with 57 cognitively intact age- and gender-matched control subjects. Mini-Mental State and Alzheimer's Disease Assessment Scale-Cognitive Subsection (ADAS-Cog) scores were determined for patients and control subjects. Aminothiols were assayed with an automated high-performance liquid chromatography (HPLC) system. RESULTS: Patients had significantly elevated total plasma homocysteine (p <.001) and cysteine (p <.01), but there were no group differences for total plasma glutathione. Glutathione was, however, a highly significant and independent predictor of cognitive scores in patients (p =.002); lower plasma levels were associated with more severe cognitive impairment. CONCLUSIONS: Total plasma homocysteine and cysteine are elevated in Alzheimer's disease, suggesting intact transsulphuration but defective remethylation of homocysteine in the disease. Total plasma glutathione levels in patients correlate with cognitive scores. Taken together, these observations perhaps reflect the differential effects of Alzheimer's disease-related oxidative stress on the two key pathways of homocysteine metabolism. PMID- 12559660 TI - Effectiveness of chromium in atypical depression: a placebo-controlled trial. AB - BACKGROUND: Chromium picolinate (CP) has been reported to benefit patients with symptoms of atypical depression. METHODS: A placebo-controlled, double-blind, pilot study of CP was conducted in 15 patients with DSM-IV major depressive disorder, atypical type. Patients received 600 micro g of CP or matching placebo (PBO) for 8 weeks. RESULTS: Seven (70%) CP and zero (0%) PBO patients met responder criteria (p =.02). Other outcomes were consistent with greater effect of CP. Three patients on CP failed to show any improvement. Chromium picolinate was well tolerated. CONCLUSIONS: Chromium picolinate shows promising antidepressant effects in atypical depression. Its mechanism of action may relate to 5HT2A downregulation, increased insulin sensitivity, or to other effects. PMID- 12559661 TI - Atypical antipsychotic drugs improve cognition in schizophrenia. PMID- 12559663 TI - Comment on "preclinical models: status of basic research in depression". PMID- 12559666 TI - Maternal serum B12 levels and risk for neural tube defects in a Texas-Mexico border population. AB - PURPOSE: Neural tube defects (NTDs) are common birth defects that can be prevented with folate fortification and supplementation. Studies suggest that other nutrients may also be essential to neural tube closure and have a potential role in risk reduction, with vitamin B(12) mentioned most often. We determined the effect of maternal serum B(12) levels, measured postpartum, on the risk of NTDs among a high risk Mexican American population. METHODS: The case-control study included 157 Mexican American women with NTD-affected pregnancies and 186 Mexican American women with normal pregnancies, who were residents of Texas Mexico border counties and delivered during 1995 to 2000. RESULTS: Compared with women in the highest vitamin B(12) quintile, women in the lowest quintile showed a strong risk effect (odds ratio (OR) = 3.0, confidence interval (CI): 1.4, 6.3); while those in the 2nd and 3rd quintiles showed moderate risk effects (OR = 1.6, CI = 0.7, 3.6 and OR = 1.7, CI = 0.8, 3.8, respectively). Adjusting for obesity, vitamin supplements, dietary folate, dietary B(12), red blood cell folate, and other covariates did not materially change these estimates. CONCLUSIONS: Insufficient levels of serum B(12), which are not normally indicative of a classical vitamin B(12) deficiency nor stem from an inadequate diet, may be an important etiologic factor for NTDs in this population. PMID- 12559667 TI - A partial SMR approach to smoothing age-specific rates. AB - PURPOSE: Age-specific rates are the fundamental measures in epidemiology. For a small population, however, rate estimates can become unstable and the age curve may contain too much random variability to adequately assess the true underlying pattern. A further problem arises when one wishes to log-transform a rate with a zero count in its numerator. The author proposes a simple and non-iterative method to stabilize rates. METHODS: The method, referred to as the "partial SMR approach," relies on finding a standard population with a similar age curve with the study population. Real and simulated data were used to demonstrate its properties. RESULTS: It is found that the choice of the standard is not critical. The method will offset automatically the role of a "dissimilar" standard; and according to the limited simulation studies, the result is still better than no smoothing. The method can also smooth the age curve adaptively, i.e., smoothing to varying degree according to internal stability of each age category. The method is asymptotically unbiased and does not have the zero-count problem. Among the various smoothing methods we have studied, the partial SMR smoothing produces the smallest mean square error and has the highest probability of successful capture of the specific pattern and trend in the age curve. CONCLUSIONS: The partial SMR smoothing is a simple and effective method for smoothing age-specific rates. PMID- 12559668 TI - The reliability of passively collected AIDS surveillance data in Massachusetts. AB - PURPOSE: To determine the reliability of demographic, clinical, and treatment information collected during routine AIDS surveillance. METHODS: Information from the medical records of a random sample of 212 AIDS cases reported to the Massachusetts Department of Public Health between November 1993 and November 1994 was compared with that from the original case reports. We assessed levels of agreement by calculating overall percent agreement and kappa statistics with 95% confidence limits. We used an intraclass correlation coefficient to compare the CD4+ lymphocyte count reported from the two sources. RESULTS: There was excellent agreement for gender (kappa = 0.97) and race (kappa = 0.87). Agreement was lower for transmission mode (kappa = 0.73), CD4+ cell count (ICC = 0.76) and category of AIDS case definition (kappa = 0.59). There was poor agreement for use of antiretrovirals (kappa = 0.23), use of prophylaxis for Pneumocystis carinii pneumonia (kappa = 0.12) and vital status (kappa = 0.22). The month and year of diagnosis agreed in only 55% of cases reviewed. CONCLUSIONS: Routine AIDS surveillance data was reliable for demographic variables, but less reliable for information about clinical events, laboratory findings, or treatment. Future AIDS surveillance efforts should improve the collection of these data by using sources where this information is more reliable. PMID- 12559669 TI - Survey non-response in the Netherlands: effects on prevalence estimates and associations. AB - PURPOSE: Differences in respondent characteristics may lead to bias in prevalence estimates and bias in associations. Both forms of non-response bias are investigated in a study on psychosocial factors and cancer risk, which is a sub study of a large-scale monitoring survey in the Netherlands. METHODS: Respondents of a cross-sectional monitoring project (MORGEN; N = 22,769) were also asked to participate in a prospective study on psychosocial factors and cancer risk (HLEQ; N = 12,097). To investigate diverse aspects of non-response in the HLEQ on prevalence estimates and associations are studied, based on information gathered in the MORGEN-project. RESULTS: A response percentage of 45% was obtained in the MORGEN-project. Response rates were found to be lower among men and younger people. The HLEQ showed a response percentage of 56%, and respondents reported higher socioeconomic status, better subjective health and healthier lifestyle behaviors than non-respondents. However, associations between smoking status and either socioeconomic status or subjective health based on respondents only were not statistically different from those based on the entire MORGEN-population. CONCLUSION: Non-response leads to bias in prevalence estimates of current smoking, current alcohol intake, and low physical activity or poor subjective health. However, non-response did not cause bias in the examined associations. PMID- 12559670 TI - Relative validity of food frequency questionnaire nutrient estimates in the Black Women's Health Study. AB - PURPOSE: The Black Women's Health Study (BWHS) was designed to investigate determinants of health and disease in US black women. More than 64,000 women are enrolled in the BWHS cohort. This study assessed the relative validity of the 68 item food frequency questionnaire (FFQ) used in the BWHS baseline questionnaire. METHODS: Four hundred and eight BWHS enrollees were asked to provide three telephone, 24-hour recalls and one written 3-day food diary over a one-year period. Means and Pearson correlations were computed to compare estimates for energy, total fat, saturated fat, protein, carbohydrate, dietary fiber, calcium, iron, vitamin C, folate, beta-carotene, and vitamin E from the FFQ, recalls, and diaries. RESULTS: Mean energy intake (kcal) was higher for the diary (1716) than the FFQ (1601) or recalls (1510). Other nutrient estimates (% kcal or per 1000 kcal) were similar across methods, except beta-carotene (FFQ higher). Correlations (energy-adjusted, except for energy, and corrected for intraperson variation) between the FFQ and the recalls were higher than for the diary data and were between 0.5 and 0.8, except for energy and vitamin E (both <0.3). CONCLUSION: The BWHS FFQ will support meaningful analyses of diet-health associations for 10 of the 11 energy-adjusted nutrient intake variables analyzed. PMID- 12559671 TI - Comparison of adipose tissue fatty acids with dietary fatty acids as measured by 24-hour recall and food frequency questionnaire in Black and White Adventists: the Adventist Health Study. AB - PURPOSE: To calibrate and compare intake of different fats and individual fatty acids as assessed with a food frequency questionnaire (FFQ) against that estimated with (i) a series of dietary recalls and; (ii) the relative fat concentration in an adipose tissue biopsy. The FFQ was specially designed for use in a cohort of Seventh-day Adventists. In preparation for a large cohort study investigating the effect of diet on risk of colon, prostate and breast cancer. METHODS: The association of adipose tissue fatty acids and dietary fat intake was assessed in 49 black and 72 white Seventh-day Adventists subjects using 8 different 24-hour recalls, a 200-item food frequency questionnaire (FFQ) and adipose tissue biopsies from each subject. RESULTS: Pearson correlation between fatty acids in adipose tissue and dietary intake as assessed by multiple 24-hour recalls were as follows: Linoleic acid: 0.77 in black and 0.71 in white subjects, respectively; Linolenic acid: 0.68 (blacks) and 0.62 (whites); Total Polyunsaturated fat (PUFA): 0.78 (blacks) and 0.70 (whites); Total Monounsaturated fat (MUFA): 0.35 (blacks) and 0.03 (whites); Total Saturated fat (SFA): 0.46 (blacks) and 0.56 (whites). Correlations between fatty acids in adipose tissue and dietary intake as assessed by FFQ were: Linoleic acid: 0.61 (blacks) and 0.52 (whites), respectively; Linolenic acid: 0.29 (blacks) and 0.49 (whites); PUFA: 0.62 (blacks) and 0.53 (whites); MUFA: 0.07 (blacks) and 0.31 (whites), SFA: 0.21 (blacks) and 0.31 (whites). CONCLUSIONS: Our study confirms findings of others that 24-hour recalls are valid for assessing dietary intake of different types of fat. The FFQ we developed and used in this study gave reasonably valid measures of fatty acid intake in our population and is thus suitable for use in large cohort studies. It had validity comparable to that observed for other FFQs. PMID- 12559672 TI - Familial aggregation of nutrient intake and physical activity: results from the San Antonio Family Heart Study. AB - PURPOSE: To evaluate the extent to which levels of physical activity and nutrient intake aggregate in families, and secondarily, to assess the repeatability of these behavioral measures over a 5-year period. METHODS: Measurements were obtained in a population-based sample consisting of 1364 members of 42 large Mexican American families. Nutrient intake was assessed by a food frequency questionnaire validated for use in this population. Usual level of physical activity was estimated using a 7-day recall questionnaire. RESULTS: Correlations between baseline (obtained 1992-1995) and follow-up (obtained 1996 to 2000) measures of all behaviors were highly significant (p < 0.001), ranging from 0.24 for % of calories derived from fat to 0.44 for % of calories derived from alcohol. Familial effects, estimated using variance component methods, were stronger when modeled as a genetic heritability than as a shared household effect; as a heritability they accounted for a significant portion of the total variation of all traits (9% for physical activity levels, p < 0.05; and 13-26% for nutrient intake, p < 0.001 for all). CONCLUSIONS: Measurements of physical activity and dietary behaviors in this population tracked over 5 years, and there was a significant degree of aggregation of these behaviors within families. Understanding the sources of these family effects may facilitate efforts to improve cardiovascular health. PMID- 12559673 TI - Factors associated with improved short term survival in obese end stage renal disease patients. AB - PURPOSE: In contrast to its role in the general population, obesity, defined as body mass index (BMI) > or = 30 kg/m(2), has been associated with improved survival in patients with end stage renal disease (ESRD). This apparent benefit has not been explained. METHODS: Using the United States Renal Data System (USRDS), we performed an historical cohort study on 151,027 patients initiated on ESRD therapy between January 1, 1995 and June 30, 1997, who never received renal transplants, and who had information sufficient to calculate BMI. We explored the association of various comorbidities present at the time of dialysis initiation (from HCFA Form 2728) with the presence of obesity by logistic regression, and the association of obesity with patient survival, including specific causes of death, by Cox regression adjusting for factors known to be associated with survival in this population. RESULTS: Obese patients had an unadjusted two-year survival of 68% compared with 58% for non obese patients. Obesity was independently associated with a reduced risk of mortality among chronic dialysis patients (adjusted hazard ratio (AHR) 0.75, 95% confidence interval, 0.72-0.78), after controlling for all comorbidities and risk factors. However, there were significantly adverse interactions among whites (AHR 1.22, 1.14-1.30, across all causes of death) and females (AHR 1.12, 1.04-1.20, entirely due to an increased risk of infectious death). CONCLUSIONS: Obesity in patients presenting with ESRD is associated independently with reduced all cause mortality; however, the relationship is complex and is stronger in African Americans. In addition, subgroup analysis suggests that obesity is associated with increased risk of infectious death in females. PMID- 12559674 TI - Acute pyelonephritis in US hospitals in 1997: hospitalization and in-hospital mortality. AB - PURPOSE: To describe the 1997 incidence of hospitalization for acute pyelonephritis in the US and the risk factors for in-hospital mortality associated with acute pyelonephritis. METHODS: Cases were defined as those with ICD9-CM codes corresponding to acute pyelonephritis in the 1997 Health Care Cost and Utilization Project (HCUP) Nationwide Inpatient Sample (NIS). Frequencies and calculations were weighted to produce either national or state estimates. Hospitalization rates were calculated using 1997 US Census Bureau population projections for the denominator. RESULTS: Females were almost five times as likely as males to be hospitalized for acute pyelonephritis (11.7/10,000 vs. 2.4/10,000), but males had higher mortality rates (16.5/1000 vs. 7.3/1000); 30% greater after adjustment. Hospitalization and in-hospital mortality rates increased with age, but not with diabetes. Mortality rates increased with number of procedures, diagnoses and having a major diagnostic category other than disorders of the kidney and urinary tract. Persons living in zip codes with lower median incomes were also at higher risk of mortality. There was little variation in mortality by hospital size, ownership, location or teaching status. CONCLUSION: In hospital mortality for pyelonephritis was associated with patient rather than hospital characteristics suggesting uniform application of standard care across hospitals and populations. PMID- 12559675 TI - A new model of isolated systolic hypertension induced by chronic warfarin and vitamin K1 treatment. AB - BACKGROUND: Isolated systolic hypertension is the predominant form of hypertension in the elderly population. Reduction of arterial compliance appears to contribute to the elevation of pulse pressure (PP) and among potential mechanisms, gradual vascular calcification, fragmentation of elastic lamellae, and augmentation of rigid component like collagen could contribute to increase aortic stiffening. Few experimental models of the disease are currently available. METHODS: To induce large artery calcification, rats were treated with warfarin and vitamin K(1) (WK) for 4 and 8 weeks, to inhibit the maturation of matrix Gla protein. The impact of chronic PP elevation was determined on large artery and cardiac remodeling and on aortic endothelial function. RESULTS: The WK treatment led to aortic medial calcification and a proportional elevation of PP, attributable mainly to a selective elevation of systolic blood pressure. The chronic treatment also increased collagen, whereas elastin decreased in the aorta. Pulse wave velocity, an index of aortic stiffening, increased in rats treated with WK. However, indices of left ventricular and aortic hypertrophy and remodeling remained normal. In addition, the WK treatment did not modify the vasoconstriction to norepinephrine and endothelin-1, and the vasodilatory response to acetylcholine and sodium nitroprusside. CONCLUSIONS: Chronic treatment with WK represents a new model of isolated systolic hypertension with several characteristics of the human disease. The relative ease to induce calcification in this model may help to foster more fundamental research, which is lacking in this type of hypertension. PMID- 12559676 TI - Angiotensin II AT1 receptor blockade improves renal perfusion in hypercholesterolemia. AB - BACKGROUND: Hypercholesterolemia (HC) is a risk factor for renal disease that may activate the angiotensin II type 1 (AT1) receptor and accelerate renal damage. Early diet-induced HC impairs renal perfusion responses, but it is yet unknown whether the AT1 receptor is involved. This study tested the hypothesis that AT1 receptor blockade improved renal perfusion and functional responses in hypercholesterolemic pigs. METHODS: Regional renal hemodynamics and function in vivo were quantified bilaterally in pigs, at baseline and during vasoactive challenge (acetylcholine or sodium nitroprusside), using electron beam computed tomography after 12 weeks of normal (n = 6) or HC diet (n = 6), or HC diet supplemented (100 mg/d) with the AT1-receptor antagonist irbesartan (HC + AT1, n = 6). RESULTS: Basal cortical and medullary perfusion was similar among the groups. Basal tubular function was similar on normal and HC diets, whereas HC + AT1 showed decreased proximal and distal fluid reabsorption. Hypercholesterolemic pigs had blunted cortical perfusion (P = .22) and augmented tubular responses to acetylcholine, whereas on HC + AT1 diet, cortical perfusion (P = .002) and tubular function were similar to normal animals. This was associated with decreased systemic levels of the oxidative stress markers thiobarbituric acid reactive substances. CONCLUSIONS: The AT1 receptor blockade in HC improves renal perfusion and tubular functional responses to endothelium-dependent vasodilators, in association with a decrease in oxidative stress. These results imply involvement of the renin-angiotensin system in the blunted renal cortical perfusion responses observed in HC, and suggest a potential role for these agents in preservation of intrarenal hemodynamics and function in HC. PMID- 12559677 TI - Effect of efonidipine and ACE inhibitors on proteinuria in human hypertension with renal impairment. AB - Although several lines of recent studies fail to demonstrate the beneficial action of calcium antagonists, a novel dihydropyridine efonidipine, which possesses dilatory action of both afferent and efferent arterioles and, therefore, shares the renal microvascular action with angiotensin converting enzyme (ACE) inhibitors, is reported to exhibit renal protection in experimental animals. The present study evaluated the effect of efonidipine and ACE inhibitors on blood pressure (BP) and proteinuria. Sixty-eight hypertensive patients with renal impairment (serum creatinine, >1.5 mg/dL) or chronic renal parenchymal disease were randomly assigned to efonidipine or ACE inhibitor treatment. Of the 68 patients, 23 were treated with efonidipine and 20 with ACE inhibitors; these patients were analyzed for the 48-week study. Both efonidipine and ACE inhibitors produced a similar degree of reductions in BP (efonidipine, from 161 +/- 2/93 +/- 2 to 142 +/- 5/82 +/- 2 mm Hg; ACE inhibitor, from 163 +/- 3/95 +/- 2 to 141 +/- 5/83 +/- 2 mm Hg), and maintained creatinine clearance for 48 weeks. Proteinuria tended to decrease in both groups, and a significant reduction was observed in proteinuric patients (>1 g/day) (efonidipine, from 2.7 +/- 0.3 to 2.1 +/- 0.3 g/day; ACE inhibitor, from 3.0 +/- 0.4 to 2.0 +/- 0.5 g/day). Of interest, efonidipine decreased proteinuria in proteinuric patients who failed to manifest decreases in systemic BP. Finally, the incidence of adverse effects, including hyperkalemia and cough, was less in the efonidipine-treated group. Both efonidipine and ACE inhibitors preserved renal function in hypertensive patients with renal impairment. The antiproteinuric effect was apparent in patients with greater proteinuria. The beneficial action of efonidipine, along with fewer side effects, may favor the use of this agent in the treatment of hypertension with renal impairment. PMID- 12559679 TI - Leukocyte angiotensin II levels inpatients with essential hypertension:relation to insulin resistance. AB - Insulin resistance is involved in the pathogenesis of type 2 diabetes, hypertension, and atherosclerosis. Angiotensin (Ang) converting enzyme inhibitors and Ang II type 1 receptor antagonists improve insulin resistance in patients with essential hypertension, which suggest that tissue Ang II is involved in insulin resistance in patients with hypertension. To investigate the participation of tissue Ang II in insulin resistance associated with hypertension, we evaluated the Ang II-generating system in leukocytes and its relation to insulin resistance in patients with essential hypertension. Eighteen patients with essential hypertension participated in this study. Ang II was separated from leukocytes by reversed-phase high-performance liquid chromatography and measured by radioimmunoassay. Insulin resistance was evaluated by determining the steady-state of plasma glucose (SSPG) concentration. The Ang I and Ang II-generating activities were evaluated in human leukocytes. Human leukocytes have Ang I- and Ang II-generating activities. The Ang II-generating activity was significantly inhibited by pepstatin A. Leukocyte Ang II level does not correlate with BP or plasma Ang II level in patients with essential hypertension. Leukocyte Ang II level strongly correlates with SSPG concentration, and significantly correlates with body mass index and plasma insulin, and with leptin levels in patients with essential hypertension. Leukocyte Ang II may be directly associated with insulin resistance. PMID- 12559678 TI - Effect of AT1 receptor blockade on endothelial function in essential hypertension. AB - BACKGROUND: Angiotensin II adversely affects endothelial function and NO availability. We analyzed the effect of AT(1) receptor blockade on endothelium dependent vasodilation and basal nitric oxide (NO) production and release in hypertensive patients. METHODS AND RESULTS: Sixty patients (53 +/- 10 years) with essential hypertension were randomized to 6 weeks of double-blind therapy with either valsartan (80 mg), hydrochlorothiazide (HCTZ) (25 mg), or placebo once daily. Basal NO production and release was assessed by measuring forearm blood flow (FBF) in response to intra-arterial infusion of N(G)-monomethyl-L-arginine (L-NMMA), and endothelium-dependent vasodilation by measuring FBF in response to intra-arterial administration of acetylcholine, respectively. Intra-arterial infusion of noradrenaline and sodium nitroprusside was used to assess endothelium independent changes in FBF. Blood pressure (BP) similarly decreased with active treatments (P < .001). After valsartan treatment, the decrease of FBF in response to L-NMMA was augmented (4 micromol/min L-NMMA, -1.3 +/- 1.2 after v -0.5 +/- 1.1 mL/min/100 mL before therapy, P < .02; 8 micromol/min L-NMMA: -1.7 +/- 1.3 after v -1.1 +/- 1.2 mL/min 100 mL before therapy, P < .05). No improvement was found after placebo or HCTZ treatment. Changes in L-NMMA-induced decrease of FBF with valsartan treatment were not related to BP changes. Neither drug substantially modified the response of FBF induced by intra-arterial infusion of acetylcholine, noradrenaline, and sodium nitroprusside. CONCLUSIONS: The AT(1) receptor blockade with valsartan improved basal NO production and release. The effect seems to be BP independent, as BP reduction with HCTZ failed to increase NO availability. PMID- 12559680 TI - Effects of nabumetone, celecoxib, and ibuprofen on blood pressure control in hypertensive patients on angiotensin converting enzyme inhibitors. AB - Nonsteroidal anti-inflammatory drugs interfere with certain antihypertensive therapies. In a double-blind study, 385 hypertensive patients stabilized on an angiotensin converting enzyme inhibitor were treated with nabumetone, celecoxib, ibuprofen, or placebo for 4 weeks. Ibuprofen caused significantly greater increases in systolic (P < .001) and diastolic (P < .01) blood pressures (BPs) compared to placebo, but not nabumetone or celecoxib. The proportion of patients with systolic BP increases of clinical concern at end point was significantly higher (P < .001) for the ibuprofen group (16.7%; 15 of 90), but not for the nabumetone group (5.5%; 5 of 91) or the celecoxib group (4.6%; 4 of 87) compared to the placebo group (1.1%; 1 of 91). PMID- 12559681 TI - Systolic blood pressure is the main etiology for poorly controlled hypertension. AB - BACKGROUND: Isolated systolic hypertension is an important risk factor for cardiovascular events. The purpose of this study was to detect the prevalence of and to evaluate the effectiveness of currently available medications in the treatment of uncontrolled isolated systolic hypertension. METHODS: We randomly selected a total of 585 patients with hypertension from our database. The two most recent blood pressure (BP) readings and other data were obtained by chart review. RESULTS: Of 585 patients, 340 (58%) had controlled BP. Of 245 patients with uncontrolled hypertension, 77.1% had uncontrolled isolated systolic hypertension and the remaining 22.9% had uncontrolled diastolic hypertension. Patients with uncontrolled systolic hypertension were on average taking more antihypertensive medications than patients with controlled BP (2.10 +/- 0.09; P = .034). CONCLUSIONS: Systolic hypertension is the etiology of uncontrolled hypertension in the majority of patients. Currently available antihypertensive medications are less effective in controlling systolic hypertension. PMID- 12559682 TI - A meta-analysis of genome-wide linkage scans for hypertension: the National Heart, Lung and Blood Institute Family Blood Pressure Program. AB - BACKGROUND: Four multicenter Networks (GenNet, GENOA, HyperGEN, SAPPHIRe) form the National Heart, Lung and Blood Institute Family Blood Pressure Program (FBPP), to search for hypertension/blood pressure (BP) genes. The networks used different family designs and targeted multiple ethnic groups, using standardized protocols and definitions. Linkage genome scans were done on samples within each network (N = 6245 relatives). METHODS: The evidence was synthesized using meta analysis. RESULTS: Combining ethnic groups, no region reached LOD >2, but several small peaks were identified, including chromosome 2p where two other recent reports find hypertension linkage. CONCLUSIONS: No regions show uniformly large effects on BP/hypertension in all populations. PMID- 12559683 TI - A genome-wide affected sibpair linkage analysis of hypertension: the HyperGEN network. AB - Results are reported here from a genome-wide linkage analysis of hypertension in a large sample of hypertensive (affected) sibpairs (650 African American and 915 white sibpairs) recruited by the HyperGEN Network of the National Heart, Lung and Blood Institute (NHLBI) Family Blood Pressure Program (FBPP). Analysis using MAPMAKER/SIBS suggests one interesting region with a LOD score of 2.08 at 63 cM from the p telomere on chromosome 2 in the African American sibpairs, which may harbor hypertension susceptibility genes. PMID- 12559684 TI - A genome-wide linkage analysis investigating the determinants of blood pressure in whites and African Americans. AB - Evidence for genomic regions influencing systolic and diastolic blood pressure (BP) were assessed in a whole genome linkage analysis in 211 African American and 160 white families as part of the GenNet network of the National Heart, Lung and Blood Institute-sponsored Family Blood Pressure Program. Multipoint regression and variance components linkage methods were used to analyze 372 polymorphic markers. Statistically compelling evidence for linkage (P values .0057 and .00023, respectively) was found on chromosome 1. Our results support the idea that BP regulation is most likely governed by multiple genetic loci, each with a relatively weak effect on BP in the population at large. PMID- 12559685 TI - Genome-wide linkage analyses for hypertension genes in two ethnically and geographically diverse populations. AB - We report the results of a genome-wide linkage scan for hypertension genes in 450 African American hypertensive sibpairs from Jackson, MS, and 539 non-Hispanic white hypertensive sibpairs from Rochester, MN. In the Jackson samples we identified one LOD score peak >1.0 on chromosome 1. In the Rochester sample, no genomic region had a LOD score >1.0. These analyses provide no appreciable evidence of hypertension genes with strong effects independent of other genetic and environmental contexts and suggest that stratified linkage analyses may be required to identify hypertension susceptibility genes in these populations. PMID- 12559686 TI - A genome scan for hypertension susceptibility loci in populations of Chinese and Japanese origins. AB - BACKGROUND: Our understanding of genes that predispose to essential hypertension is poor. METHODS: A genome-wide scan for linkage at approximately 10 cM resolution was done on 1425 sibpairs of Chinese and Japanese origins that were concordant for hypertension (N = 661), low-normal blood pressure (BP) (N = 184), or discordant for BP (N = 580). RESULTS: There was no significant evidence of linkage to a single locus in the genome. There was suggestive evidence of linkage to chromosome 10p, with a LOD score of 2.5. CONCLUSIONS: We can exclude the possibility that a single gene accounts for at least 15% of the variance in hypertension in this population. PMID- 12559687 TI - Large-scale search for genes predisposing to essential hypertension. PMID- 12559688 TI - Genome scans for hypertension and blood pressure regulation. PMID- 12559689 TI - Mechanistic and toxicokinetic data reducing uncertainty in risk assessment. AB - Risk characterization comprises hazard identification describing the intrinsic toxic potential of a chemical, knowledge of dose-response-relationships, as well as of toxicokinetics describing quantitatively the relation between external and internal dose and exposure assessment. Compounds that induce reversible effects, which are repaired during and after exposure, are considered thresholded and allow definition of a NOEL. Biological reactive intermediates of chemicals have the potential to bind covalently to cellular macromolecules like proteins and DNA. Such interaction may not be repaired completely. If damage is not repaired, the effect persists and accumulates upon repeated exposure. In such cases a NOEL cannot be determined. Thus, in the risk assessment process, data on covalent binding (CB) are of qualitative and together with toxicokinetics of quantitative significance. Qualitatively, CB, especially with DNA and in correlation with this to proteins, is indicative for an irreversible and non-thresholded mutagenic and carcinogenic effect. Absence or presence of CB assists to differentiate between primarily genotoxic and thresholded non-genotoxic carcinogens. Quantitatively, toxicokinetics together with CB are used to quantify internal exposure and target dose, which is a prerequisite for species-species extrapolation, and for extrapolation from high dose to low dose. For example, the toxicokinetics of the reactive intermediates of styrene and ethylene have been determined in rodents and humans and modeled to predict dose-responses of internal exposure. It is described in this communication that such information, together with other parameters like cell proliferation as a result of cytotoxicity, is the basis for quantitative risk assessment of human exposure to these compounds. Also for chlorobenzene, the relevance of toxicokinetics for estimating the human health risk is demonstrated. PMID- 12559690 TI - Toxicokinetic modeling and its applications in chemical risk assessment. AB - In recent years physiologically based pharmacokinetic (PBPK) modeling has found frequent application in risk assessments where PBPK models serve as important adjuncts to studies on modes of action of xenobiotics. In this regard, studies on mode of action provide insight into both the sites/mechanisms of action and the form of the xenobiotic associated with toxic responses. Validated PBPK models permit calculation of tissue doses of xenobiotics and metabolites for a variety of conditions, i.e. at low-doses, in different animal species, and in different members of a human population. In this manner, these PBPK models support the low dose and interspecies extrapolations that are important components of current risk assessment methodologies. PBPK models are sometimes referred to as physiological toxicokinetic (PT) models to emphasize their application with compounds causing toxic responses. Pharmacokinetic (PK) modeling in general has a rich history. Data-based PK compartmental models were developed in the 1930's when only primitive tools were available for solving sets of differential equations. These models were expanded in the 1960's and 1970's to accommodate new observations on dose-dependent elimination and flow-limited metabolism. The application of clearance concepts brought many new insights about the disposition of drugs in the body. In the 1970's PBPK/PT models were developed to evaluate metabolism of volatile compounds of occupational importance, and, for the first time, dose-dependent processes in toxicology were included in PBPK models in order to assess the conditions under which saturation of metabolic and elimination processes lead to non-linear dose response relationships. In the 1980's insights from chemical engineers and occupational toxicology were combined to develop PBPK/PT models to support risk assessment with methylene chloride and other solvents. The 1990's witnessed explosive growth in risk assessment applications of PBPK/PT models and in applying sensitivity and variability methods to evaluate model performance. Some of the compounds examined in detail include butadiene, styrene, glycol ethers, dioxins and organic esters/aids. This paper outlines the history of PBPK/PT modeling, emphasizes more recent applications of PBPK/TK models in health risk assessment, and discusses the risk assessment perspective provided by modern uses of these modeling approaches. PMID- 12559691 TI - Utility of physiologically based pharmacokinetic models to drug development and rational drug discovery candidate selection. AB - The present paper proposes a modeling and simulation strategy for the prediction of pharmacokinetics (PK) of drug candidates by using currently available in silico and in vitro based prediction tools for absorption, distribution, metabolism and excretion (ADME). These methods can be used to estimate specific ADME parameters (such as rate and extent of absorption into portal vein, volume of distribution, metabolic clearance in the liver). They can also be part of a physiologically based pharmacokinetic (PBPK) model to simulate concentration-time profiles in tissues and plasma resulting from the overall PK after intravenous or oral administration. Since the ADME prediction tools are built only on commonly generated in silico and in vitro data, they can be applied already in early drug discovery, prior to any in vivo study. With the suggested methodology, the following advantages of the mechanistic PBPK modeling framework can now be utilized to explore potential clinical candidates already in drug discovery: (i) prediction of plasma (blood) and tissue PK of drug candidates prior to in vivo experiments, (ii) supporting a better mechanistic understanding of PK properties, as well as helping the development of more rationale PK-PD relationships from tissue kinetic data predicted, and hence facilitating a more rational decision during clinical candidate selection, and (iii) the extrapolation across species, routes of administration and dose levels. PMID- 12559692 TI - Application of physiologically based toxicokinetic modelling to study the impact of the exposure scenario on the toxicokinetics and the behavioural effects of toluene in rats. AB - The toxicity of inhalatory exposure to organic solvents may not only be related to the total external dose, but also to the pattern of exposure. In this study physiologically based toxicokinetic (PBTK) modelling has been used to study the impact of the exposure scenario on the toxicokinetics and the behavioural effects of the model solvent toluene in rats. After construction of the model with parameters from literature, toxicokinetic data were collected from rats exposed to either a constant concentration or fluctuating concentrations at total external dose levels of 20,000 and 10,000 ppm x h for model validation. At the same exposure conditions the effects on learned performance were evaluated in separate groups of rats using a visual discrimination task. In general, the PBTK model provided reliable predictions of the toxicokinetics of toluene at different exposure scenarios, but it also tended to underestimate the blood and brain concentrations in the descending parts of the tissue concentration-time curves. At these high dose levels the differences in toxicokinetics between the constant and the fluctuating exposure groups were relatively small. The visual discrimination experiments demonstrated a slowing of response speed and disinhibition of responding in all toluene-exposed groups. The results suggest that the brain concentration of toluene is one of the major determinants of its effect on disinhibition of responding. PMID- 12559693 TI - Assessing the health risks following environmental exposure to hexachlorobutadiene. AB - Hexachloro-1,3-butadiene (HCBD) has been reported to be toxic to the rat kidney in a 2 year study at doses higher than 0.2 mg/kg/day. The toxicity is known to be a consequence of the metabolism of HCBD by glutathione conjugation and the renal beta-lyase pathway. Neither toxicity data, nor data on the metabolism of HCBD, are available in humans. In the current work, the potential of HCBD to cause kidney damage in humans environmentally exposed to this chemical has been assessed quantitatively by comparing the key metabolic steps in rats and humans. To that end, the hepatic conjugation of HCBD with glutathione, the metabolism of the cysteine conjugate by renal beta-lyases and N-acetyltransferases, and the metabolism of the N-acetylcysteine conjugate by renal acylases has been compared in vitro in rat and human tissues. Rates for each metabolic step were lower in humans than in rats; 5-fold for glutathione conjugation, 3-fold for beta-lyase and 3.5-fold for N-acetyltransferase. Acylase activity could not be detected in human kidney cytosol. Use of these data in a physiologically based toxicokinetic model to quantify metabolism by the beta-lyase pathway demonstrated that metabolism in humans was an order of magnitude lower than that in rats. At the no effect level for kidney toxicity in the rat the concentration of beta-lyase metabolites was calculated by the model to be 137.7 mg/l. In humans the same concentration would be achieved following exposure to 1.41 ppm HCBD. This is in contrast to the figure of 0.6 ppb which is obtained when it is assumed that the risk is associated with the internal dose of HCBD itself rather than beta-lyase metabolites. PMID- 12559695 TI - Physiologically-based kinetic modeling of vapours toxic to the respiratory tract. AB - The respiratory tract is frequently identified as a site of toxicity for inhaled xenobiotic chemicals. Usually, these observations come from controlled animal studies. For these studies to be of quantitative value to human health risk assessment, species-specific factors governing dosimetry of inhaled substances must be taken into account. Toxicokinetics of vapours in the respiratory tract are defined by absorption, distribution, metabolism, and excretion, as they are in other tissues; however, these concepts take on new dimensions when considering respiratory tract toxicants, especially those that elicit portal of entry effects by directly interacting with the tissue lining the respiratory tract. Species specific factors related to anatomy, physiology and biochemistry govern inter species extrapolation of toxicokinetics. This article discusses critical factors of respiratory tract kinetics that should be considered when developing physiological-based toxicokinetic (PBTK) models for inhaled vapours. Important considerations such as impact of regional airflow-delivery, water solubility, reactivity, and rates of local biotransformation on respiratory tract tissue dosimetry are highlighted. These factors can be accounted for only to a limited extent when using default approaches to extrapolate dosimetry of inhaled substances across species. On the other hand, PBTK modeling has the flexibility to accommodate many of the critical determinants of respiratory tract toxicity. PBTK models can also help identify the most critical toxicokinetic data necessary to replace defaults. PBTK approaches have led to more informed estimates of human target tissue dose, and therefore human health risk, especially where these risk assessments have been based on extrapolation of animal dosimetry studies. Experience derived from the development of more intensive case studies have, in turn, enabled simplified approaches to the use of PBTK modeling for respiratory tract toxicants. Whether simplified or highly complex, PBTK modeling approaches are proven to be of great utility to risk assesors interested in applying quantitative information to informed risk assessment evaluations. PMID- 12559694 TI - A toxicokinetic model for styrene and its metabolite styrene-7,8-oxide in mouse, rat and human with special emphasis on the lung. AB - Styrene (ST) occurs ubiquitously in the environment and it is an important industrial chemical. After its uptake by the exposed mammalian organism, ST is oxidized to styrene-7,8-oxide (SO) by cytochrome P450 dependent monooxygenases. This reactive intermediate is further metabolized by epoxide hydrolase (EH) and glutathione S-transferase (GST). In long-term animal studies, ST induced lung tumors in mice but not in rats. Considering the lung to be the relevant target organ for ST induced carcinogenicity in mice, we extended a previously developed physiological toxicokinetic model in order to simulate the lung burden with ST and SO in the ST exposed mouse, rat and human. The new model describes oral and pulmonary uptake of ST, its distribution into various tissues, its exhalation and its metabolism to SO in lung and liver. It also simulates the distribution of the produced SO into the tissues and its EH and GST mediated metabolism in liver and in lung. In both organs the ST induced GSH consumption is described together with the formation of adducts to hemoglobin and to DNA of lymphocytes in ST exposed mice, rats and humans. The model includes compartments for arterial, venous and pulmonary blood, liver, muscle, fat, richly perfused tissues and lung. The latter organ is represented by two compartments, namely by the conducting and the alveolar zone. The physiological description of the pulmonary compartments relies on measured alveolar retentions, literature values of surface area of capillary endothelium, of the thickness of the tissue 'air-to-plasma', of the partition coefficient lung:blood and of metabolic parameters of ST and SO measured in pulmonary cell fractions of rodents and humans. Simulations of average pulmonary GSH levels in ST exposed rodents agree with measured data. The model predicts a significant GSH depletion (40%) in the conducting zone of mice exposed for 6 h to a ST concentration of only 20 ppm. In the conducting zone of rats, exposure to 200 ppm ST results in a loss of GSH of about 15% only. In humans, a pulmonary GSH reduction does not occur. The highest average pulmonary SO concentrations are predicted for mice, somewhat lower values for rats and by far the lowest ones for humans. Following steady state exposure to 20 ppm ST, the average SO concentration in mouse lungs is expected to be only three times higher than in rats. This difference diminishes to a factor of less than two at 70 ppm. In humans exposed to 20 ppm ST for 8 h, the average pulmonary SO burden of 0.016 micromol/kg is predicted to be about 17 and 50 times smaller than the corresponding values for rat and mouse. In agreement with reported values, pulmonary DNA adduct levels in rodents exposed to 160 ppm ST were simulated to be similar in rats and mice. In summary, there was no dramatic difference in the calculated average pulmonary SO burden between both animal species. However, pulmonary GSH loss was by far more expressed in ST exposed mice than rats. Since the model was validated on all available ST/SO data in mice, rats and humans, we consider it to be useful for estimating the risk resulting from exposure to ST. PMID- 12559696 TI - Risk assessment dosimetry model for inhaled particulate matter: I. Human subjects. AB - Pollutant particulate matter (PM) is a serious global problem, presenting a threat to the health and well being of human subjects. Inhalation exposures tests with surrogate animals can be performed to estimate the threat. However, it is difficult to extrapolate the findings of animal tests to human conditions. In this two-part series, interspecies dosimetry models especially designed for implementation with risk assessment protocols are presented. In Part I, the mathematical integrity of the source model per se was tested with data from human subjects, and theoretical predictions agreed well with experimental measurements. In Part II, for surrogate (rat) simulations, appropriate algorithms for morphologies and ventilatory parameters were used as subroutines in the validated model. We conducted a comprehensive series of computer simulations describing the behavior of a representative air pollutant, secondary cigarette smoke. For risk assessment interests, a range of states from rest to exercise was considered. PM hygroscopicity had a pronounced effect on deposition in a complex but systematic manner, in humans and rats: deposition was increased for particles larger than about 1 microm, but was decreased for particles smaller than about 0.1 microm. The results clearly indicate that dosimetry models can be effectively used to a priori determine the laboratory conditions necessary for animals tests to accurately mimic human conditions. Moreover, the use of interspecies models is very cost effective. We propose, therefore, that mathematical models be used in a complementary manner with inhalation exposure experiments and be actively integrated into PM risk assessment protocols. PMID- 12559697 TI - Risk assessment dosimetry model for inhaled particulate matter: II. Laboratory surrogates (rat). AB - Inhalation toxicology investigations are often performed with laboratory animals to address the potential health effects of inhaled air pollutants on human beings. In Part II of this risk assessment study we have considered the deposition of inhaled particulate matter in the laboratory rat as the surrogate of choice. Calculations were performed in an analogous manner to those conducted in Part I for human subjects. To simulate a wide range of human respiratory intensities associated with different levels of physical activities that must be recognized in the determination of air pollution standards, the CO(2) concentrations within animal inhalation exposure chambers may be controlled. Accordingly, we have regulated rat breathing parameters to correspond to a range of human activities, from rest to work. The results of this interspecies modeling study have been presented in a variety of graphical formats to ease comparisons with findings from experiments and to facilitate integration of the results into risk assessment analyses. The findings of our work clearly demonstrate that interspecies simulations can be employed to design animal tests a priori so that the results can be effectively and efficiently extrapolated to human conditions in a meaningful manner. PMID- 12559698 TI - The Bayesian population approach to physiological toxicokinetic-toxicodynamic models--an example using the MCSim software. AB - The calibration of physiologically based toxicokinetic models against experimental data encompasses the merging of prior knowledge with information present in the data. This prior knowledge is manifested in the scientific literature and associated with various degrees of uncertainty. The most convenient way to combine these sources of information is via the use of Bayesian statistical methods. Furthermore, toxicokinetic models are subject to both inter- and intra-individual variability. This variability may be handled statistically by the use of a population model. The MCSim software, which is available for free download on the Internet, permits the use of a population model in combination with a Bayesian statistical approach. An example of the use of MCSim in a recent model-based risk assessment of dichloromethane (DCM) is given and discussed. PMID- 12559699 TI - Practical application of kinetic data in risk assessment--an IPCS initiative. AB - In this paper, guidance developed in a project of the International Programme on Chemical Safety (IPCS) initiative on Harmonization of Approaches to the Assessment of Risk from Exposure to Chemicals is considered in the context of its application in the assessment of the adequacy of physiological-toxicokinetic (PTK) modeling to inform quantitatively extrapolations for interspecies differences and human variability in dose-response assessment. This guidance was developed in the context of a framework, which permits the incorporation of quantitative chemical-specific data, relating to either toxicokinetics or toxicodynamics to replace part or all of the usual 100-fold default uncertainty factor for interspecies differences or human variability in the development of tolerable or reference doses or concentrations. However, since the guidance relates specifically to adequacy of kinetic or dynamic data to replace default for interspecies and human variability, it is also applicable to other approaches of dose-response analyses such as estimation of cancer potency or risk. The framework also supports probabilistic characterization, where data are sufficient. This guidance has been developed and refined through a series of planning and technical meetings and larger workshops, in which a broad range of participants from academia, government agencies, and the private sector have prepared and gained experience in application through case studies. The guidance for adequacy of kinetic data to replace default is presented in the context of several categories, including determination of the active chemical species, choice of the appropriate kinetic parameter and experimental data, the latter which includes reference to relevance of population, relevance of route, relevance of dose/concentration, and adequacy of number of subjects/samples. The principal objective of this guidance, which has been developed primarily as a resource for risk assessors, is to foster better understanding of the criteria for adequacy of chemical-specific data to quantitate interspecies differences and human variability in kinetics and dynamics, including PTK models. It is anticipated that the guidance will also encourage generation of appropriate data and models, and facilitate their incorporation in dose/concentration-response assessment for regulatory purposes. In this paper, the application of the guidance is considered primarily through reference to examples, with emphasis on those where PTK models have been informative. PMID- 12559700 TI - The integration of data on physico-chemical properties, in vitro-derived toxicity data and physiologically based kinetic and dynamic as modelling a tool in hazard and risk assessment. A commentary. AB - Toxicity of a compound for an organism is dependent on the route of exposure, the amount (or concentration), the way in which the compound is taken up, distributes and is eliminated from the organism (ADME, kinetics) and the intrinsic properties (reactivity; mode of action, dynamics) of the compound towards the organism. These three elements: exposure, kinetics and dynamics form the basis of hazard and risk evaluations. Developments in our knowledge of the way in which physico chemical properties of chemicals (on the one side) and physiological processes in the organism (on the other side) determine a compound's toxicity have greatly increased our understanding of toxicological processes and our ability to interpret experimental results. This has now resulted in the development of model systems in which the above-mentioned processes can be described mathematically. Biokinetic modelling is currently of great interest, but the further development of toxicodynamic modelling is equally important. The combination of both allows the estimation of a compound's critical amount/concentration on the critical site of action, which ideally would be the basis for hazard and risk assessments. In vitro systems have been extremely useful in studying the molecular basis of a chemical's biological activity, including its mechanism(s) of toxic action. Other achievements include the prediction of biological reactivity on the basis of a compound's physico-chemical properties and the construction of quantitative structure-activity relationships (QSARs). However, for the incorporation of in vitro-derived data as well as the results of QSARs, kinetic modelling is indispensable. Thus, biokinetic and toxicodynamic modelling are important (if not crucial) tools in toxicological research and there are increasing opportunities to incorporate the results of this work in hazard and risk assessments. Their implementation will allow a much more scientifically-based and a better structured risk assessment, which will be to a much lesser extent relying on animal experimentation. PMID- 12559701 TI - Panel discussion: application of physiological-toxicokinetic modelling. PMID- 12559707 TI - Camel trypanosomosis in Morocco 1: results of a first epidemiological survey. AB - An epidemiological survey of camel trypanosomosis was conducted for the first time in Morocco in 1997 and 1998. Five provinces located in the south and southeast of the High Atlas mountain chain were included in this study. A total number of 1460 serum samples were collected and tested by a card agglutination test for trypanosomosis (CATT) and enzyme-linked immunosorbant assay (Ab-ELISA) to detect anti-Trypanosoma evansi antibodies. The overall seroprevalence was 14.1% by CATT and 18.2% by Ab-ELISA. Two very active foci were identified in Zagora and Merzouga with high Ab-ELISA prevalences of 43.3 and 35.4%, respectively, for each area. The concordance between the CATT and Ab-ELISA was 94.1%. The age-related distribution of seroprevalence showed a tendency for the infection rate to increase with age up to a maximum in the 7-10 year-old group. Clinical examination revealed that enlargement of lymph nodes was the most frequent sign in seropositive animals (79.7%). The comparison of haematocrit values of seropositive and seronegative animals showed a significant difference that indicated severe anaemia in infected animals. Isolation of the parasite by inoculation of mice allowed us to collect 15 T. evansi isolates from Zagora and three from Merzouga. The study showed that the Saharan provinces of Morocco were slightly affected by camel trypanosomosis but two hyperendemic foci, Zagora and Merzouga, were found to be sources of contamination for the surrounding regions. PMID- 12559708 TI - Diagnosis and quantification of Theileria sergenti using TaqMan PCR. AB - Theileria sergenti is the causative agent of persistent theileriosis in cattle. The ubiquitous infection of theileiriosis causes chronic anemia and fever in cattle, especially in exogenous cattle. In this study, we applied real-time polymerase chain reaction (PCR) for the diagnosis and quantification of parasite using specific primers for 33 kDa gene fragment of T. sergenti. Comparison of TaqMan PCR with traditional microscopic method, Giemsa's staining, on blood collected from cattle revealed the specificity up to 0.00005% of parasitemia to traditional diagnosis. In addition, it was found that this method can estimate the relative status of infection among herds. The results of present study showed that this method is not only applicable to detect the chronic infection of Theileria, but also effective in evaluation on parasitemia status of cattle, thus it can be used in monitoring the health status in field. PMID- 12559709 TI - Molecular and morphologic characterization of a Cryptosporidium genotype identified in lemurs. AB - This study reports the molecular and morphologic characterization of a Cryptosporidium sp., identified in stools of captive lemurs Propithecus verreauxi coquereli. Stool samples were collected from seven animals (n=7) presenting episodes of diarrhea. Bright-field light microscopy of stool smears stained with modified acid-fast technique revealed the presence of Cryptosporidium sp. oocysts in four of the stool samples analyzed. All microscopically positive samples were confirmed by PCR using primers designed to amplify DNA fragments from two independent loci, i.e. the Cryptosporidium oocyst wall protein (COWP) gene and the small subunit ribosomal RNA (ssrRNA) gene. Phylogenetic analysis based on the full-length ssrRNA gene placed this isolate within a clade that contains all currently known C. parvum species/genotypes, closely related to the C. parvum pig genotype. Comparison with partial ssrRNA sequences available in the GenBank revealed 100% sequence identity with the genotype previously identified in Canadian patients. This finding was confirmed further by comparison of the COWP gene partial sequences. PMID- 12559711 TI - Kinetics of circulating antigens in pigs experimentally infected with Taenia solium eggs. AB - Three groups of four piglets were experimentally infected with different doses (10(3), 10(4) and 10(5)) of Taenia solium eggs whereas a fourth group of two pigs received gravid proglottids. At autopsy 6 months post infection, the two latter pigs were heavily infected with more than 3000 living cysts per kg of muscle. Ten of the 12 other pigs harboured light infections, i.e. between 2 and 107 cysticerci, 42.4% of which were degenerated. The two remaining pigs had no detectable cysts at post mortem examination. Circulating antigens (CA) were detected in the sera of all pigs harbouring living cysticerci using a monoclonal antibody based ELISA. CA were first detected between 2 and 6 weeks post infection and remained present generally throughout the entire observation period even in pigs carrying only five to eight living cysts, although strong fluctuations of the level of CA were observed in some pigs. In animals without living cysts at post mortem CA were only detected for a short period and disappeared presumably when the cysticerci became degenerated. The minimum number of living cysts, which could be detected using this ELISA, was 1. PMID- 12559710 TI - Evaluation of tongue inspection and serology for diagnosis of Taenia solium cysticercosis in swine: usefulness of ELISA using purified glycoproteins and recombinant antigen. AB - Evaluation of serology using glycoproteins (GPs) purified by preparative isoelectric focusing (pH 8.8) and recombinant chimeric antigen (RecTs) of Taenia solium was carried out using (1) blood samples on filter papers from pigs infected with different doses of eggs of T. solium in Mexico, (2) serum samples from pigs found infected naturally in Vietnam and Ecuador and (3) serum samples from pigs suspected to be infected with T. solium by tongue inspection in Tanzania. Antibody responses (IgG) were detectable in experimentally infected pigs confirmed harbouring 16 or more cysts at necropsy from 30 days after egg inoculation. One of three pigs naturally infected and harbouring 2.5 cysts/kg muscle and most of pigs harbouring=5.0 cysts/kg were also seropositive by ELISA. Although pigs may be infected with other taeniid species such as Taenia hydatigena, pigs harbouring this parasite were negative in ELISA. Approximately, 76 and 78% of sera from pigs having nodule(s) in the tongue (positive tongue inspection) were serologically positive by both ELISA and immunoblot, respectively. Furthermore, approximately 34 and 18% of sera from pigs having no nodules in the tongue (negative tongue inspection) were also seropositive by ELISA and immunoblot, respectively. ELISA using the two antigens was more sensitive than immunoblot and reliable for differentiation of pigs infected with cysticerci of T. solium from those either uninfected or infected with other taeniid species. Pigs without nodule by tongue inspection should be checked serologically in endemic areas. PMID- 12559712 TI - Experimental haemonchosis in goats: effects of single and multiple infections in the host response. AB - Histopathological changes and the distribution of T lymphocytes (CD3), B cells (CD79alpha) and IgG secreting plasma cells were recorded in the abomasum and abomasal lymph nodes of goats during early and late post-infection stages with one to four doses of Haemonchus contortus L3. The infiltration of eosinophils, mast cells, CD3(+) T lymphocytes, CD79alpha(+)B cells and IgG(+) plasma cells in the abomasal mucosa increased dramatically from 10dpi onwards, whereas globule leukocytes were observed only during chronic infection. In late post-infection stages abomasal infiltration of globule leukocytes, CD3(+) T lymphocytes, CD79alpha(+)B cells and IgG(+) plasma cells was significantly higher (P<0.05) in reinfected (groups 6-8) than in primarily infected goats (group 5). In the abomasal lymph nodes, marked hyperplasia of lymphoid follicles and medullary cords, with increase of CD3(+) T lymphocytes, CD79alpha(+)B cells and IgG(+) plasma cells was recorded from 10dpi (group 3) onwards. Worm burdens and the severe abomasal response during the late post-infection stages suggests that a rapid expulsion of nematodes did not occur. The prolonged time required for generating globule leukocytes suggested that immune mechanisms dependent of this cell type are of crucial importance in the protective immunity against H. contortus in goats. PMID- 12559713 TI - Assessment of cattle grub (Hypoderma spp.) prevalence in northeastern Italy: an immunoepidemiological survey on bulk milk samples using ELISA. AB - Hypodermosis is a myiasis of cattle mainly found in the Northern Hemisphere. In Italy it has been reported primarily in southern regions with a farm prevalence reaching a peak of 85%, while few data are available in the north of the country. This paper reports the results of a serological survey carried out on hypodermosis in Vicenza province (northeastern Italy). During January and February 1999, bulk milk samples were collected from 224 dairy farms and tested, using an hypodermosis ELISA kit. Epidemiological data, such as farm location, herd size, number of cows, grazing management, treatments against Hypoderma larvae, presence of imported animals, were gathered and statistically analysed in relation to the serological results. In 15 farms the presence of warbles was monitored by manual palpation and larvae collected for identification.Ninety seven (43.3%) samples were detected as positive, whereas 12 (5.4%) and 115 (51.3%) were inconclusive and negative, respectively. Thirteen farms were positive upon parasitological examination and the larvae collected were identified as Hypoderma bovis.Seroprevalence was highly correlated with the practice of free grazing and slightly, but significantly, correlated with the number of cows on the farm. The mean antibody level of positive herds was significantly higher on farms without imported animals. On farms with warbled animals there was a high inverse correlation between antibody level and quantity of milk in the tanks and the number of lactating cows. The ELISA test on bulk milk samples has been shown, under field conditions, to be a useful, cost effective and easy test for the diagnosis of bovine hypodermosis. PMID- 12559714 TI - Rotenone and oxalic acid as alternative acaricidal treatments for Varroa destructor in honeybee colonies. AB - This experiment assessed the efficacy of rotenone and oxalic acid (OA) in an aqueous sugar solution in controlling the honeybee mite Varroa destructor. Colonies were populated with mite-infested brood combs and worker bees. Three rotenone or OA treatments administered during the period with capped brood on 31 July, 14 and 18 August resulted in an average efficacy of 24.10%. In untreated colonies mite mortality averaged 5.40%. No significant differences (P>0.05) were found between the rotenone and OA treatments. Three OA treatments administered on 9, 12 and 18 September resulted in a 77.93% mite mortality. An increase in mite drop (P<0.05) was observed at 2 and 4 days after each treatment. OA applications in broodless colonies resulted in significantly (P<0.001) higher mite mortality rates (98.65% average) than the three treatments of rotenone or OA in colonies with capped brood. The dynamics of mite mortality after each rotenone or OA treatment are discussed in this study. PMID- 12559715 TI - Survey of Theileria lestoquardi antibodies among Sudanese sheep. AB - The prevalence of Theileria lestoquardi antibodies in Sudanese sheep from nine geographical areas in Sudan was determined using indirect fluorescent antibody "IFA" test. Out of 315 samples examined, 51 (16.2%) were found positive and ranged between 23.4% in River Nile State and 10% in Kasala and Darfour Provinces with an overall prevalence of 16.2% indicating widespread distribution of the infection. We also report on presence of antibodies reactive to Theileria annulata in sheep sera. PMID- 12559716 TI - Characteristics of oestrus measured using visual observation and radiotelemetry. AB - The aim of this study was to measure the level of agreement between characteristics of oestrus measured using visual observation and radiotelemetry. Oestrous cycles in 20 non-lactating Holstein cows were synchronised by treatment with an intravaginal progesterone releasing inserts and injections of oestradiol benzoate for a first oestrus (Round 1) and then re-synchronised for two subsequent and successive oestrous cycles (Rounds 2 and 3). Cows were ovariectomised following the third synchronised oestrus and then retreated to induce oestrus (Round 4). Oestrus was monitored by continuous visual observation and radiotelemetry between 24 and at least 60 h after removal of inserts at Rounds 1-4. Significant differences (P<0.001) were obtained between the mean number of mounts (42.7 versus 17.8), duration of oestrus (14.4h versus 10.9h), total duration of mounts (223.9s versus 33.1s), mean duration per mount (5.6s versus 1.9s), the number of mounts per hour (3.5 versus 2.0) and the interval to oestrus (41.2h versus 42.8h) between values determined by visual observation compared with radiotelemetry, respectively. The 95% limits of agreement for all the characteristics of oestrus measured were wide although the efficiency of detection of oestrus for both methods of recording oestrus were high (97.5% (78/80) versus 93.8% (75/80), for visual observation and radiotelemetry, respectively; P=0.25). We conclude that there is poor agreement between characteristics of oestrus measured with visual observation and radiotelemetry in Holstein cows following monitoring of synchronised oestrous cycles, although both methods were equally efficient for the detection of oestrus. PMID- 12559717 TI - Effect of oestradiol benzoate given after prostaglandin at two stages of follicle wave development on oestrus synchronisation, the LH surge and ovulation in heifers. AB - Oestrus synchronization following prostaglandin-induced luteolysis is variable and dependent on follicle wave status in cattle. Oestradiol benzoate (ODB) has been used following prostaglandin to reduce the interval to oestrus and ovulation, but the effect of follicle wave status at the time of ODB administration is not clear. The aim of this study was to characterize the endocrine and follicular responses following ODB after luteolysis at different stages of the follicle wave. Prostaglandin was administered at either emergence or dominance of the second follicle wave. Twenty-four hours later animals received either 0.5mg ODB in oil or a control oil injection. Follicular development was monitored daily by ultrasonography, oestrous behavior was determined and blood samples were collected. In animals treated with ODB at emergence, there was a reduction (P<0.05) in the maximum diameter of the ovulatory follicle (11.7+/-1.2 mm versus 13.1+/-0.1 mm) and in the interval from prostaglandin to oestrus (52.0+/-2.3 h versus 88.0+/-9.6h), to the LH surge (53.3+/-3.5 h versus 89.1+/-6.5 h) and to ovulation (96+/-0.0 h versus 129.6+/ 9.6h), compared with controls. In animals treated with ODB at dominance, there was a reduction (P<0.05) in the interval from prostaglandin to the LH surge (54.0+/-3.1 h versus 70.9+/-4.8 h), but not in the interval from prostaglandin to oestrus (53.3+/-2.7 h versus 65.7+/-4.5 h; P=0.11), to ovulation (96.0+/-0.0 h versus 110.4+/-4.8 h; P=0.12) or the maximum diameter of the ovulatory follicle (12.7+/-0.3 mm versus 13.6+/-0.4 mm; P=0.12), compared with controls. Treatment did not affect (P>0.05) the length of the subsequent oestrous cycle or corpus luteum size. In conclusion, the use of ODB advanced, but did not alter the temporal relationships among oestrus, the LH surge and ovulation, regardless of stage of follicle development at treatment. PMID- 12559719 TI - Meiotic competence of bovine fetal oocytes following in vitro maturation. AB - The in vitro ability between fetal and cow oocytes to resume meiosis and progression to metaphase-II (M-II) was compared. Cumulus oocyte complexes (COCs) were harvested from 2 to 6 mm follicles from ovaries of 7.5 month to term fetuses and adult cows. Cumulus cells were removed using 3 mg/ml hyaluronidase and repeated pipetting. Denuded oocytes were fixed in 3% glutaraldehyde, stained with DAPI and evaluated under fluorescent microscopy for nuclear status before in vitro maturation (IVM). COCs from fetal and adult ovaries were also matured in 200 microl droplets of medium 199 supplemented with 10 microg/ml FSH, 10/ml LH, 1.5 microg/ml estradiol, 75 microg/ml streptomycin, 100 IU/ml penicillin, 10 mM hepes and 10% FBS for 24 h at 39 degrees C and 5% CO(2). Matured oocytes were fixed, stained and evaluated as explained above for nuclear status namely stage of germinal vesicle (GV) development and subsequent meiotic competence. Data were analyzed using chi-square analysis. The majority of fetal oocytes (P<0.05) before IVM were at GV stages GV-I (27.7%), GV-II (37.6%) and GV-V (22.8%) compared to cow oocytes, which were at GV stages IV (28.3%) and V (46.7%). After IVM, fewer fetal oocytes were at earlier stages of GV development and majority (P<0.05) were at GV-V (24.0%), premetaphase (17.4%) and metaphase-I (M-I: 7.2%) stages. However, after IVM, more cow oocytes matured to M-II than did fetal oocytes (93.7% versus 26.9%; P<0.05). In conclusion, fetal oocytes do not mature in vitro as well as cow oocytes. Our findings suggest that the low meiotic competence of fetal oocytes can be attributed to their being at earlier stages of GV development before IVM. PMID- 12559718 TI - Ultrasound image characteristics of ovarian follicles in relation to oocyte competence and follicular status in cattle. AB - Assessment of the quality of the female gamete has become paramount for in vitro procedures. There is a need to identify reliable indicators of oocyte competence and develop a simple, non-invasive method to assess competence. The aim of this study was to investigate the relationships among ultrasonographic attributes of a follicle, its stage of development and the competence of the oocyte that it contains. We tested the hypotheses that follicular echotexture characteristics are related to: (1) the phase of development of the follicle, (2) the presence of the corpus luteum (CL) and/or the dominant follicle in the ovary, and (3) developmental competence of cumulus oocyte complexes (COC) from the same ovary. Crossbred beef cows (n=143), age 4-14 years, were given a luteolytic dose of dinoprost to cause ovulation. Ultrasound-guided ablation of all follicles > or = 4mm was done 8 days later to induce new follicular wave emergence during a luteal phase. Ultrasonographic images of dominant follicles and the three largest subordinate follicles (n=402 follicles; 84 cows) were acquired on Days 2, 3, 5 or 7 of the follicular wave (Day 0: wave emergence), i.e. growing, early-static, late static, and regressing phases of subordinate follicle development, respectively. From a subset of these animals (n=33), ovaries were collected within 30 min of slaughter and COC from subordinate follicles > or = 3mm underwent in vitro maturation, fertilization and culture to the blastocyst stage.Image analysis revealed differences in echotexture between dominant and subordinate follicles among Days 2-7 of the follicular wave. Images of dominant and subordinate follicles at Day 7 of the wave displayed consistently lower grey scale values (P<0.05) in the peripheral antrum, follicular wall and perifollicular stroma than all other days. Follicle images displayed a consistent pattern of variation in echotexture among follicular phases. Data did not support the hypothesis of a local effect of the CL or dominant follicle on follicular echotexture. Echotexture values of the perifollicular stroma were lower in ovaries that did not produce embryos compared to ovaries that produced embryos. Our results showed that the changes in follicular image attributes are consistent with changes in follicular status. The sensitivity of the technique is not yet sufficient for use in a diagnostic setting, but results provide rationale for further development of image analysis as a tool for evaluating oocyte competence in situ. PMID- 12559720 TI - The effect of dietary protein restriction on the secretion of LH and FSH in pre pubertal female lambs. AB - The effect of restricted dietary protein on the synthesis, storage and release of LH and FSH was studied in pre-pubertal female lambs. The experiment started when the lambs were aged 12 weeks and weighed 26.0+/-1.6 kg. It was conducted for 25 weeks. The lambs were fed isocaloric diets containing either a restricted level of crude protein (8% CP; n=6; treatment R) or an elevated one (18% CP; n=4; treatment E). At 37 weeks of age and before the first oestrous cycle, blood samples were collected over 6 h at 10 min intervals for LH assay. The lambs were slaughtered and their brains recovered and fixed in situ. Immuno-reactive (IR) LH and FSH cells were localised by immunohistochemistry techniques. Messenger RNA analyses used by non-isotope in situ hybridisation with sense and anti-sense riboprobes from beta subunits of LH and FSH cDNA clones. Data were generated using computer analysis to measure the proportion of IR and/or hybridising cells and their optical density for immuno-staining and hybridisation signal. Plasma LH was measured by RIA. The daily live-weight gains were 56.5+/-13.1 g and 97.8+/ 14.3 g for R and E lambs, respectively (P<0.05), so that final weights at slaughter were 36.1+/-1.97 kg and 39.1+/-3.44 kg, respectively (P<0.05). The number of cells expressing LH beta mRNA and the optical density of this hybridisation signal was significantly (P<0.001) lower in the R lambs but the number of IR LH positive cells was higher (P<0.001) than for the E lambs. The concentration of LH in the plasma of R sheep was lower (P<0.05) than the E group and this response was associated with a decrease (P<0.05) in LH pulse frequency and amplitude. Dietary protein concentration appeared to have no effect on the IR in FSH cells or on the expression of FSH beta mRNA. In summary, the low protein diet influenced the body weight and weight gain of growing lambs and exerted an inhibitory effect on the synthesis and the release of LH in the pituitary gonadotrophs. No such effect was observed for FSH. It was concluded that the protein concentration of the diet consumed during the growth of female lambs may be an important modulator of processes leading to the pre-pubertal rise in LH. PMID- 12559721 TI - Effect of diet and GnRH administration on post-partum ovarian cyclicity in autumn lambing ewes. AB - Using autumn-lambing ewes, this study investigated (i) the effects of diet on gonadotrophin secretion and responsiveness of the hypothalamic-pituitary-ovarian axis to exogenous GnRH during the early post-partum period; and (ii) whether ovulation prior to completion of uterine involution results in an increased incidence of aberrant ovarian cycles. Thirty-two ewes rearing 1.9+/-0.12 lambs were equally allocated to two dietary treatments at lambing (22 October +/-0.2 day). Diets comprised ad libitum hay and 1.5 kg per ewe per day of one of two concentrates (11.5 MJ ME, 195 g CP per kg) containing 300 g kg(-1) cracked maize grain (M) or 300 g kg(-1) sugar beet pellets (S). Half of the ewes on each diet (G) received 25 i.v. injections of 250 ng GnRH in 2 ml 0.9% saline at 2 h intervals from days 12-14 post-partum while remaining ewes (N) were monitored for the resumption of spontaneous ovarian cyclicity. Blood samples were obtained from all ewes throughout the study (lambing to 18 December) for measurement of circulating hormone concentrations and the uteri and ovaries of all ewes were examined via laparoscopy on day 21 post-partum. There were no effects of dietary treatment on ewe daily live weight loss, lamb daily live weight gain or the immediate post-partum increase in circulating FSH concentrations. Diet did not affect insulin concentrations or LH pulse frequency on day 12 post-partum but LH pulse amplitude was lower in ewes fed concentrate M compared to concentrate S (1.4+/-0.10 versus 1.7+/-0.12 ng ml(-1), respectively, P<0.05) and this was associated with an increased interval to the resumption of spontaneous ovarian cycles (35+/-3.1 versus 26+/-2.1 day, respectively, P<0.05). Administration of exogenous GnRH increased (P<0.05) the proportion of ewes on both diets that ovulated within 20 days of parturition and advanced the onset of ovarian cyclicity in ewes fed concentrate M by 9.5 days (significance of interaction, P<0.05). Four ewes, all of which ovulated before day 22 post-partum, had extended luteal activity while in remaining ewes, duration of the first luteal phase was inversely related to the time of first ovulation (r(2)=0.16, P<0.05). Results demonstrate that (i) the onset of ovarian cyclicity is influenced by diet and can be advanced by administration of exogenous GnRH; and (ii) ovulation during the early post-partum period is associated with an increased incidence of extended luteal activity. PMID- 12559722 TI - hCG treatment on day of mating improves embryo viability and fertility in ewe lambs. AB - An investigation was conducted to determine the effects of supplementing hCG at mating time on luteal function, conceptus growth, placentation and reproductive performance in TexelxClun Forest ewe lambs. After oestrus synchronisation with progestagen sponges and PMSG ewe lambs were treated either with normal saline (n=102) or 150 IU of hCG (n=105) at mating time. At 25 days after mating 24 animals were slaughtered from each group to determine embryo viability and placental development. hCG significantly (P<0.05) increased crown-rump length (saline: 11.9+/-0.2 mm; hCG: 12.7+/-0.2 mm), amniotic sac width (saline: 11.4+/ 0.4 mm; hCG: 12.0+/-0.3 mm) and the number of placentomes (saline: 90.8+/-7.3; hCG=122.4+/-6.3). Among the pregnant animals that were slaughtered on 25 days post-mating, ovulation rate tended to be higher in the hCG group compared to controls (saline: 1.16; hCG: 1.54). However, no difference was observed either in mean plasma progesterone concentrations (saline: 4.6 ng/ml; hCG: 4.9 ng/ml; sed 0.56) or in progesterone production from luteal slices when cultured in vitro (saline: 239.6+/-11.8 ng/mg; hCG: 263.2+/-13.6 ng/mg) between controls and hCG treated animals. Reproductive performance was also recorded in ewe lambs that were either treated with saline (n=78) or hCG (n=81). The total number of lambs born (saline: 38; hCG: 58) was significantly (P<0.05) greater in hCG group compared to saline-treated controls. Both lambing percentage (saline: 36%; hCG: 48%) and litter size (saline: 1.35; hCG: 1.48) tended to be greater (P<0.10) in hCG-treated animals compared to the controls. In conclusion, these data suggest that treatment of ewe lambs with hCG at the time of mating improves conceptus growth, placentation and number of lambs born. PMID- 12559723 TI - Relationship between intrafollicular concentrations of parathyroid hormone related peptide (PTHrP) and steroid hormones in oestrogenic and non-oestrogenic ovarian follicles in the mare. AB - The objective of the present study was to determine whether parathyroid hormone related peptide (PTHrP) is present in the equine follicular fluid and if so, how it is related to the follicular development in the horse. For this purpose, ovaries were collected from 40 Thoroughbred and Thoroughbred Cross mares at slaughter during the period from February to May. Normal growing follicles were dissected from the ovaries of each mare and their diameters measured. A total of 174 follicles was used in this study. The follicular fluid was aspirated from each follicle and assayed for PTHrP, oestradiol (E), testosterone (T) and progesterone (P). The follicles were classified as either oestrogenic or non oestrogenic if the follicular fluid content of oestradiol was >40 or <40 ng/ml, respectively. PTHrP concentrations were significantly (P<0.05) higher in oestrogenic follicles, but T and P concentrations did not differ. Furthermore, E:T ratio was significantly (P<0.05) greater in oestrogenic follicles compared to the non-oestrogenic ones. The mean diameter of oestrogenic follicles was significantly (P<0.05) greater than that of non-oestrogenic ones. The higher concentrations of PTHrP observed in the follicular fluid of healthy oestrogenic follicles suggest that it may have a role in the control of ovarian function. PMID- 12559724 TI - Effect of ascorbic acid and Vitamin E supplementation on semen quality and biochemical parameters of male rabbits. AB - The objective of this study was to determine the effects of supplementation of ascorbic acid, Vitamin E (Vit. E) and their combination in drinking water on sperm characteristics, lipid peroxidation (LPO) and seminal plasma enzymes of mature male rabbits. Twenty-four male New Zealand White rabbits (5 months old) were given drinking water supplemented with ascorbic acid (1.5 g/l), Vit. E (1.0 g/l) and ascorbic acid+Vit. E (1.5+1.0 g/l) for 12 weeks. Vitamin supplementation in drinking water increased feed intake, but body weight gain was not significantly affected. Concentrations of thiobarbituric acid-reactive substances (TBARS) were significantly (P<0.05) reduced in seminal plasma of treated groups compared with the control. Treatment with ascorbic acid, Vit. E, and their combination significantly (P<0.05) increased lipido (reaction time), ejaculate volume, sperm concentration, total sperm output, sperm motility index, total motile sperm, packed sperm volume, initial hydrogen ion concentration (pH), and semen initial fructose concentration. Abnormal and dead sperm were significantly (P<0.05) decreased in treated animals. The activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) were significantly (P<0.05) decreased, whereas glutathione S-transferase (GST) showed a significant increase in seminal plasma of treated animals compared with the controls. The results from this study indicated that supplementation of drinking water with antioxidant ascorbic acid, Vit. E and their combination reduced the production of free radicals and can improve rabbit semen quality, but the greater improvement seemed to be from Vit. E. PMID- 12559725 TI - Vitrification of in vitro cultured rabbit morulae. AB - In the present work, we attempt to establish an efficient vitrification procedure for 32-cell rabbit embryos obtained in vitro. In experiment 1, both the effect of the composition of the vitrification solutions and the cryoprotectant addition (either in one or two steps) were studied. For one-step addition, straws with embryos in the final vitrification solution (total time 60s) were plunged into liquid nitrogen. For two-step addition, previously embryos were 2 min pre equilibrated in 0.5 ml of (1:1) PBS plus 20% FCS: vitrification solution without sucrose. Different solutions of cryoprotectants were compared: 25 vol.% ethylene glycol supplemented with 0.25 M sucrose (25EG+S) and 20% ethylene glycol plus 20% dimethyl sulfoxide, alone (20EG+20DMSO-S) or supplemented with 0.25 M sucrose (20EG+20DMSO+S). Six percent (30/487) of the total of 32-cell embryos obtained by in vitro culture in each experimental session was slow-frozen by a classical method as a technical efficiency control. Only 30% slow-frozen embryos reached blastocyst stage. Significant differences in embryo development were detected between the one-step (25EG+S) and two-step (25EG+S) groups and the one-step (20EG 20DMSO+S) and two-step (20EG-20DMSO-S) groups (0-6% versus 36-50%, respectively). Consequently, in the following experiments only these two vitrification procedures were used. In experiment 2, we attempted to substitute the use of PBS by HEPES-buffered Ham's F-10 (h-CM) in all cryoprotective solutions or media. When h-CM was used, a significant reduction in the in vitro embryo development was observed when the HEPES-buffered groups were compared with one-step (20EG 20DMSO+S) group in s-PBS (35-45% versus 73%). In experiment 3, the one-step (20EG+20DMSO+S) and two-step (20EG+20DMSO-S) procedures were assayed using two FCS levels (20 and 40%) in the PBS-based media. Relative to in vitro development, the highest rates were reached with one step (20EG-20DMSO+S), using PBS plus 20% FCS, which was different from two steps (20EG-20DMSO-S), regardless of percentage of FCS in the PBS-based media (81% versus 41-45%; P<0.05). In conclusion, we propose either the one step (20EG-20DMSO+S) or two steps (20EG-20DMSO-S) prepared in PBS plus 20% serum for use in future works. PMID- 12559727 TI - Synthesis of 1,2,3-tri-O-acetyl-5-deoxy-D-ribofuranose from D-ribose. AB - A practical route towards the synthesis of 1,2,3-tri-O-acetyl-5-deoxy-D ribofuranose from D-ribose is described. The key steps include deoxygenation of methyl 2,3-O-isopropylidene-5-O-sulfonyloxy-beta-D-ribofuranoside by reductive displacement employing hydride reagents. Subsequent total hydrolysis followed by acetylation led to the title compound in 56% overall yield from D-ribose. The sequence is simple, inexpensive, high yielding and clearly suitable for multi gram preparations. PMID- 12559728 TI - Remote control of alpha- or beta-stereoselectivity in (1-->3)-glucosylations in the presence of a C-2 ester capable of neighboring-group participation. AB - In (1-->3)-glucosylation the glycosyl bond originally present in either donor or acceptor is shown to control the stereoselectivity of the forthcoming bond, i.e., the newly formed glycosidic linkage has the opposite anomeric configuration of that of either the donor or acceptor. Therefore, with alpha-(1-->3)-linked disaccharides with nonreducing ends that have the 3-OH free as the acceptor and an acetylated glucosyl trichloroacetimidate as the donor, or with an alpha-(1- >3)-linked acetylated disaccharide trichloroacetimidate as the donor and a glucoside with 3-OH free as the acceptor, beta-linked trisaccharides were obtained. Meanwhile, with beta-(1-->3)-linked disaccharides that have nonreducing ends with the 3-OH free as the acceptor and an acetylated glucosyl trichloroacetimidate as the donor, or with a beta-(1-->3)-linked acetylated disaccharide trichloroacetimidate as the donor and a glucoside with the 3-OH free as the acceptor, alpha-linked trisaccharides were obtained in spite of the C-2 neighboring group participation. PMID- 12559729 TI - Enzymatic synthesis of beta-xylanase substrates: transglycosylation reactions of the beta-xylosidase from Aspergillus sp. AB - A beta-D-xylosidase with molecular mass of 250+/-5 kDa consisting of two identical subunits was purified to homogeneity from a cultural filtrate of Aspergillus sp. The enzyme manifested high transglycosylation activity in transxylosylation with p-nitrophenyl beta-D-xylopyranoside (PNP-X) as substrate, resulting in regio- and stereoselective synthesis of p-nitrophenyl (PNP) beta-(1- >4)-D-xylooligosaccharides with dp 2-7. All transfer products were isolated from the reaction mixtures by HPLC and their structures established by electrospray mass spectrometry and 1H and 13C NMR spectroscopy. The glycosides synthesised, beta-Xyl-1-->(4-beta-Xyl-1-->)(n)4-beta-Xyl-OC6H4NO2-p (n=1-5), were tested as chromogenic substrates for family 10 beta-xylanase from Aspergillus orizae (XynA) and family 11 beta-xylanase I from Trichoderma reesei (XynT) by reversed-phase HPLC and UV-spectroscopy techniques. The action pattern of XynA against the foregoing PNP beta-(1-->4)-D-xylooligosaccharides differed from that of XynT in that the latter released PNP mainly from short PNP xylosides (dp 2-3) while the former liberated PNP from the entire set of substrates synthesised. PMID- 12559730 TI - Structure of the O-specific polysaccharide of Proteus vulgaris O45 containing 3 acetamido-3,6-dideoxy-D-galactose. AB - An O-specific polysaccharide was isolated by mild acid degradation of the lipopolysaccharide of Proteus vulgaris O45 and studied by sugar and methylation analyses along with 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, ROESY, H-detected 1H,13C HSQC and HMBC experiments. The following structure of the pentasaccharide repeating unit of the polysaccharide was established:-->6)-alpha D-GlcpNAc-(1-->4)-alpha-D-GalpNAc-(1-->4)-alpha-D-GalpA-(1-->3)-beta-D-GlcpNAc-(1 ->2)-beta-D-Fucp3NAc4Ac-(1-->where Fuc3NAc4Ac is 3-acetamido-4-O-acetyl-3,6 dideoxygalactose. A cross-reactivity of anti-P. vulgaris O45 serum was observed with several other Proteus lipopolysaccharides, which contains Fuc3N derivatives. PMID- 12559731 TI - Relationships between the molecular structure and moisture-absorption and moisture-retention abilities of carboxymethyl chitosan. II. Effect of degree of deacetylation and carboxymethylation. AB - Carboxymethyl chitosans (CM-chitosan) of various degrees of deacetylation (DD 28 95%) and substitution (DS 0.15-1.21) were successfully prepared from N acetylchitosans in NaOH of varying concentrations. Infrared spectroscopy (IR), elemental analysis, potentiometric titration, 13C NMR, X-ray diffraction and gel permeation chromatographic (GPC) techniques were used to characterize their molecular structures. The moisture-absorption (R(a)) and -retention (R(h)) abilities of CM-chitosan are closely related to the DD and DS values. Under conditions of high relative humidity, the maximum R(a) and R(h) were obtained at DD values of about 50%, and when the DD value deviated from 50%, R(a) and R(h) decreased. Under dry conditions, when the DD value was 50%, the R(h) was the lowest. With the DS value increasing, R(a) and R(h) increased. However, further increase of the DS value above 1.0 reduced the increasing tendency of R(a) and R(h), and even some decreases in R(a) and R(h) were observed. Intermolecular hydrogen bonds play a very important role in moisture-absorption and retention ability of CM-chitosan. PMID- 12559732 TI - Primary structure of the 2-O-methyl-alpha-L-fucose-containing side chain of the pectic polysaccharide, rhamnogalacturonan II. AB - A 2-O-methylfucosyl-containing heptasaccharide was released from red wine rhamnogalacturonan II (RG-II) by acid hydrolysis of the glycosidic linkage of the aceryl acid residue (AceA) and purified to homogeneity by size-exclusion and high performance anion-exchange chromatographies. The primary structure of the heptasaccharide was determined by glycosyl-residue and glycosyl-linkage composition analyses, ESIMS, and by 1H and 13C NMR spectroscopy. The NMR data indicated that the pyranose ring of the 2,3-linked L-arabinosyl residue is conformationally flexible. The L-Arap residue was confirmed to be alpha-linked by NMR analysis of a tetraglycosyl-glycerol fragment, [alpha-L-Arap-(1-->4)-beta-D Galp-(1-->2)-alpha-L-AcefA-(1-->3)-beta-L-Rhap-(1-->3)-Gro], generated by Smith degradation of RG-II. Our data together with the results of a previous study,(1) establish that the 2-O-Me Fuc-containing nonasaccharide side chain of wine RG-II has the structure (Api [triple bond] apiose): [see structure]. Data are presented to show that in Arabidopsis RG-II the predominant 2-O-MeFuc-containing side chain is a mono-O-acetylated heptasaccharide that lacks the non-reducing terminal beta L-Araf and the alpha-L-Rhap residue attached to the O-3 of Arap, both of which are present on the wine nonasaccharide. PMID- 12559733 TI - Crystallographic analysis of the thermal motion of the inclusion complex of cyclomaltoheptaose (beta-cyclodextrin) with hexamethylenetetramine. AB - The crystal structure of the inclusion complex of cyclomaltoheptaose (beta cyclodextrin) with hexamethylenetetramine was determined at temperatures of 123, 173, 223, and 293 K. The rigid-body motion of the host and guest molecules was evaluated by means of the TLS method that represents the molecular motion in terms of translation, libration, and screw motion. In increasing the temperature from 123 to 293 K, the amplitude of the rigid body vibration of the host molecule was increased from 1.0 to 1.3 degrees in the rotational motion and from 0.16 to 0.17 A in the translational motion. The cyclomaltoheptaose molecule has the flexibility in seven alpha-(1-->4)-linkages, and each glucose unit was in the rotational vibration around an axis through two glycosidic oxygen atoms. As a result, the rigid-body parameters of cyclomaltoheptaose were considered to be overestimated because of including the contribution from the local motion of glucose units. In contrast, for the guest molecule having no structural flexibility, the TLS analysis demonstrated that the atomic thermal vibration was mostly derived from the rigid body motion. The rotational amplitude of hexamethylenetetramine was changed from 5.2 to 6.6 degrees in increasing the temperature from 123 to 293 K, while the change of the translational amplitude was from 0.20 to 0.23 A. The translational motion of the guest molecule was hindered by the inside wall of the host cavity. The molecular motion was characterized by the rotational vibration around the axis through two nitrogen atoms that were involved in the hydrogen-bond formation. PMID- 12559734 TI - Dissolution of sucrose crystals in the anhydrous sorbitol melt. AB - The dissolution of a sugar (sucrose as a model) with higher melting point was studied in a molten food polyol (sorbitol as a model) with lower melting point, both in anhydrous state. A DSC and optical examination revealed the dissolution of anhydrous sucrose crystals (mp 192 degrees C) in anhydrous sorbitol (mp 99 degrees C) liquid melt. The sucrose-sorbitol crystal mixtures at the proportions of 10, 30, 60, 100 and 150 g of sucrose per 100 g of sorbitol were heat scanned in a DSC to above melting endotherm of sorbitol but well below the onset temperature of melting of sucrose at three different temperatures 110, 130 and 150 degrees C. The heat scanning modes used were with or without isothermal holding. The dissolution of sucrose in the sorbitol liquid melt was manifested by an increase in the glass transition temperature of the melt and corresponding decrease in endothermic melting enthalpy of sucrose. At given experimental conditions, as high as 25 and 85% of sucrose dissolved in the sorbitol melt during 1 h of isothermal holding at 110 and 150 degrees C, respectively. Optical microscopic observation also clearly showed the reduction in the size of sucrose crystals in sorbitol melt during the isothermal holding at those temperatures. PMID- 12559736 TI - Beta-anomeric selectivity in the glycosidation of D-mannofuranurono-6,3-lactone catalyzed by boron trifluoride diethyl etherate. AB - The BF3-promoted glycosylation of D-mannofuranurono-6,3-lactone with dodecanol or methanol afforded n-alkyl beta-D-mannofuranosidurono-6,3-lactone. Reduction of n dodecyl beta-D-mannofuranosidurono-6,3-lactone with sodium borohydride yielded the corresponding alkyl beta-D-mannofuranoside. PMID- 12559735 TI - Remote site bromination via a cascade rearrangement involving two bridging dioxonium species during oxidative cleavage of a benzylidene acetal. AB - Attempted cleavage of the benzylidene group of 3,5-O-benzylidene-2,6,7-tri-O trimethylacetyl-D-glycero-D-gulo-heptono-1,4-lactone with N-bromosuccinimide led to the formation of a 7-bromo derivative with a benzoyl group in the 3-position and trimethylacetyl groups in the 5 and 6 positions. Analysis of the coupling constants in the proton NMR spectra indicated that both trimethylacetyl groups had participated to shift the crowded electron-deficient center formed at the 5 position by the decomposition of the bromobenzylidene group to the much more accessible 7-position. The net result of this cascade rearrangement was retention at both the C-5 and C-6 positions. This is an interesting example of a cascade rearrangement under strict entropic and stereo control the synthetic utility of which is being investigated. PMID- 12559737 TI - Rapid quantification of O-acetyl and O-methyl residues in pectin extracts. AB - A rapid method for the determination of the degrees of methylation (DM) and acetylation (DA) of pectins was developed. The polymer substitution degree as determined after saponification at 80 degrees C with NaOD during 1H NMR analysis. Under alkaline conditions, the cleavage of O-acetyl and O-methyl linkages allows the detection and the integration of the H-4 signal from galacturonic acid residues in the newly unesterified pectins. So, after a 10-min NMR recording, sodium acetate and sodium methanolate can be easily quantified relative to the clearly identified H-4 signal in galacturonic acid residues. Protons signals from pectin neutral sugars do not interfere with H-4. During the analysis, a limited (<3%) methanol evaporation leading to a weak reduced signal from the methanolate protons was observed. The proposed method allows in few minutes an accurate simultaneous quantification of DM and DA from few mg of pectin extracts, without the need of external standards. PMID- 12559738 TI - The structure of the O-specific polysaccharide chain of the Shewanella algae BrY lipopolysaccharide. AB - An acidic O-specific polysaccharide was obtained by mild acid degradation of the Shewanella algae strain BrY lipopolysaccharide and was found to contain L rhamnose, 2-acetamido-4-[D-3-hydroxybutyramido)]-2,4,6-trideoxy-D-glucose (D BacNAc4NHbu), and 2-amino-2,6-dideoxy-L-galactose, N-acylated by the 4-carboxyl group of L-malic acid (L-malyl-(4-->2)-alpha-L-FucN) in the ratio 2:1:1. 1H and 13C NMR spectroscopy was applied to the intact polysaccharide, and the following structure of the repeating unit was established:-3)-alpha-D-BacNAc4NHbu-(1-->3) alpha-L-Rha-(1-->2)-alpha-L-Rha-(1-->2)-L-malyl-(4-->2)-alpha-L-FucN-(1-. The repeating unit includes linkage via the residue of malic acid, reported here for the first time as a component of bacterial polysaccharides. PMID- 12559740 TI - Molecular dynamics simulations of an oligosaccharide using a force field modified for carbohydrates. AB - Parameterization of the phi and omega torsion angles in pyranosidic saccharides was performed based on density functional theory calculations. The modified CHARMM force field, which is referred to as PARM22/SU01, was tested on a glucosyl trisaccharide. A molecular dynamics simulation of the oligosaccharide with explicit water as solvent was performed to investigate the conformational flexibility. Protonz.sbnd;proton distances and heteronuclear spin-spin coupling constants were calculated from the trajectories and showed good agreement to those previously determined by NMR spectroscopy. PMID- 12559741 TI - Synthesis of new sugar amino acid derivatives of D-glucosamine. AB - The synthesis of several new sugar amino acid derivatives of 2-acetamido-2-deoxy D-glucose, bearing a C-glycosyl functionality as building blocks for the design and synthesis of natural glycoconjugates mimetics, is described. These compounds were prepared from the readily accessible per-benzylated amino C-allyl glucopyranosyl compounds, with TMSOTf/Ac(2)O-mediated selective acetolysis of the 6-O-benzyl group as the key step. PMID- 12559742 TI - Convenient approach to higher carbon sugars. First synthesis of the free C12 sugar: D-erythro-L-manno-D-manno-dodecose. AB - The model synthesis of a C12-aldose was initiated from the easily available dimethyl(benzyl 2,3,4-tri-O-benzyl-alpha-D-manno-heptopyranos-6-ulos-7 yl)phosphonate and 2,3:4,5-di-O-isopropylidene-D-arabinose. PMID- 12559743 TI - Characterizing the pH-dependent stability and catalytic mechanism of the family 11 xylanase from the alkalophilic Bacillus agaradhaerens. AB - The xylanase, BadX, from the alkalophilic Bacillus agaradhaerens was cloned, expressed and studied in comparison to a related family 11 xylanase, BcX, from B. circulans. Despite the alkaline versus neutral conditions under which these bacteria grow, BadX and BcX both exhibit optimal activity near pH 5.6 using the substrate o-nitrophenyl beta-xylobioside. Analysis of the bell-shaped activity profile of BadX yielded apparent pK(a) values of 4.2 and 7.1, assignable to its nucleophile Glu94 and general acid Glu184, respectively. In addition to having an approximately 10-fold higher k(cat)/K(m) value with this substrate at pH 6 and 40 degrees C, BadX has significantly higher thermal stability than BcX under neutral and alkaline conditions. This enhanced stability, rather than a shift in its pH optimum, may allow BadX to hydrolyze xylan under conditions of elevated temperature and pH. PMID- 12559744 TI - Isolation and characterisation of the homogalacturonan from type II cell walls of the commelinoid monocot wheat using HF-solvolysis. AB - In contrast to the typical type I cell wall of the dicot plants, the type II cell wall of the commelinoid monocot plants is known to be relatively poor in pectins. Assuming a critical role for the remaining pectins in terms of cell wall architecture and/or as a reservoir of signalling molecules, we have compared different protocols for the isolation of the main pectin polymer, homogalacturonan, from wheat leaf cell walls. Pectin was detected in these cell walls immunochemically using the monoclonal antibodies JIM5 and JIM7, and biochemically by monosaccharide analysis. The Ca(++)-chelators CDTA and imidazole extracted a pectin rich fraction from isolated cell walls which was however contaminated with significant amounts of hemicelluloses. Pretreatment of the cell walls with anhydrous hydrogen fluoride at controlled low temperatures followed by HF/ether- and water-extraction prior to imidazole-extraction of pectins yielded a purer homogalacturonan fraction. The near absence of rhamnosyl residues proved that the isolated homogalacturonan fraction was free of rhamnogalacturonans. If HF-solvolysis was performed at -23 degrees C, the resulting homogalacturonan had a degree of methyl esterification identical to that of the pectins in the initial wheat cell wall. The antibodies JIM5 and JIM7 as well as PAM1 and LM5 proved that the isolated homogalacturonan had a low methyl ester content, was polymeric and free of galactan side chains. We can thus isolate native homogalacturonan from the type II wheat cell walls with the original in muro pattern of methyl esterification still intact, to further investigate e.g., its degradability by plant or microbial pectic enzymes. PMID- 12559745 TI - Effect of molecular structure on the conductivity of amorphous carbohydrate-water KCl mixtures in the supercooled liquid state. AB - The effect of carbohydrate structure on the conductivity of low water content amorphous carbohydrate-water, and carbohydrate-water-KCl mixtures, has been measured using both direct current and alternating current techniques at temperatures in the supercooled liquid and glassy range, ranging from -40 to 80 degrees C. The structures included homologous mono-, di- and trisaccharides (glucose, maltose and maltotriose), a monosaccharide with no exocyclic hydroxymethyl group (xylose) and a second trisaccharide (raffinose). The KCl mixtures contained 9.3% w/w water and 0.74% w/w KCl which resulted in calorimetric glass transition temperatures, T(g), in the range -29-19 degrees C. At this concentration conduction due to KCl dominated that due to intrinsic conductors originating from the carbohydrates and water. In the supercooled liquid region, as temperature, T, is reduced to T(g), the activation energy of the molar conductivity of KCl, Lambda(m), increased as described by a Vogel Tamman-Fulcher-type equation, Lambda(m)=Lambda(m0)exp[B/(T-T(0))], where Lambda(m0), B and T(0) are constants. Comparison of the molar conductivity of KCl in the carbohydrate mixtures at T(g) with that in aqueous solutions showed that conductivity is, to varying extents, uncoupled from viscosity. The uncoupling increased in the order D-xylose3)-alpha-L-6dTalp4Ac-(1- >3)-beta-D-Galp-(1-->7)-alpha-Kdop-(2-->. PMID- 12559751 TI - Structure and physicochemical properties of starches from kidney bean seeds at immature, premature and mature stages of development. AB - Starches from kidney bean (Phaseolus vulgaris L. cv. Toramame) seeds at the immature, premature, mature stages of development were examined. The starch content increased from 94, 219 to 265 mg per seed. Starches showed the C(a) crystalline type composed of small (<5 micrometer) and large (10-35 micrometer) granules, with the large granules largely increasing with maturity. The amylose content increased from 21, 26 to 27%, and rapid viscograms and DSC thermograms suggested that the mature-stage starch was gelatinized with ease. The amylose increased in size from DPn 820, 1000 to 1080 and a number of chains per molecule (NC) from 3.3, 4.2 to 4.5. The branched amylose was a minor component (11-18% by mole) with NC 20-22. The amylopectin was similar in CL (23), beta-amylolysis limit (59%), and chain-length distribution, but reduced in size (DPn 17,100-5270) and increased in content of phosphorus (114-174 ppm) with an increase in the amount of phosphorus linked to C-6 of the glucose residue (8-66%). PMID- 12559752 TI - Examination of the structure of amylopectin molecules by fluorescent labeling. AB - Amylopectin molecules from rice, maize, sweet potato and potato were examined by fluorescent labeling followed by gel-permeation HPLC. The number-average degree of polymerization (dp(n)) was determined to be in range of 9600-15,900. The molar based distribution revealed the presence of three molecular species, large (dp(n) 13,400-26,500), medium (4400-8400) and small (700-2100). Their molar proportions differed by plant origin. The large species was a major component (43-63% by mole). A relatively large amount of the medium (16-28% by mole) and small (19 38%) species was found although their weight proportion was small (8-15, 1-4%, respectively). The three species from waxy rice amylopectin had a similar chain length distribution and also a similar size-distribution of C chains. These results suggested that the three species were basically similar in cluster structure but different in number of clusters per molecule. PMID- 12559754 TI - Vesicle budding: a coat for the COPs. AB - Although vesicular transport in eukaryotic cells involves a number of different carriers, one common feature is that most of them use small GTPases to direct coat assembly at the donor membrane. COPII coated vesicles bud from the endoplasmic reticulum to selectively export secretory cargo en route to the Golgi complex. Vesicle formation involves the stepwise recruitment of the small GTPase Sar1 and two large heterodimeric complexes Sec23-Sec24 and Sec13-Sec31 to the membrane. A new structural study now provides breathtaking molecular insights into the formation of the Sec23-Sec24-Sar1 pre-budding complex and into COPII coat assembly. PMID- 12559755 TI - Npap60: a new player in nuclear protein import. AB - Until very recently, the vertebrate protein Npap60/Nup50 was thought merely to be a component of the nuclear pore complex (NPC). This conclusion was based on the observations that Npap60/Nup50 localizes at the NPC by immunofluorescence and electron microscopy and also contains FG (Phe-Gly) repeats, a motif commonly found in nucleoporins but not in proteins located elsewhere. However, far from being a fixed structural component of the NPC, it now appears as though Npap60 can shuttle from one side of the NPC to the other. Most significantly, a recent paper shows that Npap60 enhances the nuclear import of a cargo possessing a basic nuclear localization sequence by associating directly with the import cargo carrier complex and (presumably) moving through the NPC with it. Several NPC proteins have now been shown to be mobile in the NPC, and this new report might indicate that these 'mobile' nucleoporins play a more active role in the nuclear transport of cargo than was previously appreciated. PMID- 12559756 TI - New roles for p21 and p27 cell-cycle inhibitors: a function for each cell compartment? AB - Cell division relies on the activation of cyclins, which bind to cyclin-dependent kinases (CDKs) to induce cell-cycle progression towards S phase and later to initiate mitosis. Since uncontrolled cyclin-dependent kinase activity is often the cause of human cancer, their function is tightly regulated by cell-cycle inhibitors such as the p21 and p27 Cip/Kip proteins. Following anti-mitogenic signals or DNA damage, p21 and p27 bind to cyclin-CDK complexes to inhibit their catalytic activity and induce cell-cycle arrest. Interestingly, recent discoveries suggest that p21 and p27 might have new activities that are unrelated to their function as CDK inhibitors. The identification of new targets of Cip/Kip proteins as well as evidence of Cip/Kip cytoplasmic relocalization have revealed unexpected functions for these proteins in the control of CDK activation, in the regulation of apoptosis and in transcriptional activation. This article discusses recent insights into these possible additional functions of p21 and p27. PMID- 12559757 TI - Intramembrane-cleaving proteases: controlled liberation of proteins and bioactive peptides. AB - This review summarizes recent findings and common principles for intramembrane cleaving proteases that catalyse critical steps in cell regulation and signalling and which are involved in diseases such as Alzheimer's disease and hepatitis C virus infection. PMID- 12559758 TI - Insulin/IGF and target of rapamycin signaling: a TOR de force in growth control. AB - 'They come in all sizes.' Apart from its origin and use in the clothing industry, this saying reflects the fact that the size of organisms spans an enormous range. Whether destined to be large or small, species grow in an organized fashion to reach their final specified size. For growth to proceed, food must be metabolized to liberate energy in the form of adenosine triphosphate (ATP) and protein building blocks in the form of amino acids. One major orchestrator of this complex growth process in diverse metazoan species is the insulin/insulin-like growth factor (IGF) system. This review summarizes current studies primarily from Drosophila regarding the function of the insulin/IGF system in the control of growth. PMID- 12559760 TI - Caveolae: anchored, multifunctional platforms in the lipid ocean. AB - The function of caveolae is hotly debated. It now seems clear that caveolae are stable membrane domains that are kept in place by the actin cytoskeleton. However, this stability can be perturbed, leading to caveolar internalization. Caveolae are important in the regulation of various signaling processes, such as nitric oxide activity, and in cholesterol efflux and cholesterol-ester uptake. Caveolin deficiency particularly affects the cardiovascular system and the lungs but, because the knockout mice are viable, none of the proposed functions appears to be essential. Rather than having a specific function, caveolae might be considered to be multifunctional organelles with a physiological role that varies depending on cell type and cellular needs. PMID- 12559759 TI - Branching morphogenesis of the lung: new molecular insights into an old problem. AB - It has been known for decades that branching morphogenesis of the lung is mediated through reciprocal interactions between the epithelium and its underlying mesenchyme. In recent years, several key players, in particular members of the major signaling pathways that mediate this interaction, have been identified. Here, we review the genetic and molecular studies of these key components, which have provided a conceptual framework for understanding the interactions of these major signaling pathways in branching morphogenesis. The future challenge is to translate understanding of the signaling cascade into knowledge of the cellular responses, including cell proliferation, migration and differentiation, that lead to the stereotyped branching.* PMID- 12559761 TI - Fluorescence imaging of signaling networks. AB - Receptor-triggered signaling processes exhibit complex cross-talk and feedback interactions, with many signaling proteins and second messengers acting locally within the cell. The flow of information in this input-output system can only be understood by tracking where and when local signaling activities are induced. Systematic strategies are therefore needed to measure the localization and translocation of all signaling proteins, and to develop fluorescent biosensors that can track local signaling activities in individual cells. Such a biosensor tool chest can be based on two types of green fluorescent protein constructs that either translocate or undergo fluorescence-resonance-energy transfer when local signaling occurs. Broad strategies to measure quantitative, dynamic parameters in signaling networks, together with perturbation approaches, are needed to develop comprehensive models of signaling networks*. PMID- 12559762 TI - Interfering with leukocyte rolling--a promising therapeutic approach in inflammatory skin disorders? AB - Many novel anti-inflammatory compounds have been derived from research tools designed to better understand the complex process of leukocyte extravasation at sites of inflammation. Given that selectin-mediated rolling is the crucial initial step of leukocyte localization to the skin, specific interference with this mechanism is a particularly attractive target to treat cutaneous inflammation. Recently, several exciting advances have been reported aimed at selective interference with specific target molecules crucially involved in leukocyte rolling. PMID- 12559764 TI - Using genome-wide mapping in the mouse to identify genes that influence drug response. AB - Differential drug response is most often likely to be a complex trait, controlled by the combined influences of multiple genes and environmental influences. As a result of theoretical and technical limitations, to date, most clinically useful pharmacogenomic studies in humans have been limited to a small number of candidate genes that have a relatively major impact on drug response. Here, the problems involved in identifying genes that underlie drug response in humans are discussed and the power of mouse genetics as a tool for pharmacogenomic discovery is highlighted. PMID- 12559763 TI - Characterization of the UDP-glucose receptor (re-named here the P2Y14 receptor) adds diversity to the P2Y receptor family. AB - The cloning of a human G-protein-coupled receptor (GPCR) that specifically responds to UDP-glucose and related sugar-nucleotides has been reported recently. This receptor has important structural similarities to known members of the P2Y receptor family but also shows a distinctly different pharmacological response profile. Here, the IUPHAR Subcommittee for P2Y receptor nomenclature and classification review the current knowledge of this receptor and present their reasons for including this receptor in the P2Y receptor family as the P2Y(14) receptor. PMID- 12559765 TI - Crystallization of the human glucocorticoid receptor ligand binding domain: a step towards selective glucocorticoids. AB - The crystal structure of the glucocorticoid receptor (GR) ligand binding domain in a ternary complex with dexamethasone and a TIF2 coactivator peptide has been determined recently. The structure reveals several distinct features not found in other nuclear receptors, such as a novel dimerization interface and a second charge clamp that might be important in determining coactivator binding selectivity. The GR ligand binding domain also has a steroid binding pocket that is distinct from other nuclear receptors and might explain its selectivity for glucocorticoids and its diversity of responses. PMID- 12559766 TI - Do unpredictable toxic side-effects of drugs reflect their promiscuous binding energies? PMID- 12559767 TI - Pharmacological effects of garlic extract. PMID- 12559768 TI - Empirical models and Hill coefficients. PMID- 12559769 TI - Too much of a good thing: adenosine overload in adenosine-deaminase-deficient mice. AB - Chronic lung diseases are associated with persistent lung inflammation and damage. The mechanisms that govern the chronic nature of these disorders are not known. Adenosine is a signaling nucleoside that is generated in hypoxic environments such as that found in the inflamed lung, which suggests that it might serve a regulatory role in chronic lung diseases. Support for this hypothesis comes from studies in adenosine-deaminase-deficient mice where lung adenosine levels accumulate in association with increased lung inflammation and damage. Furthermore, lowering adenosine levels or antagonizing adenosine receptors can reverse pulmonary phenotypes in this model, suggesting that chronic adenosine elevations can affect signaling pathways that mediate aspects of chronic lung disease. PMID- 12559770 TI - Acquired somatic mutations in the molecular pathogenesis of COPD. AB - Chronic obstructive pulmonary disease (COPD) is caused mostly by cigarette smoking but its specific molecular mechanisms are obscure. Current theories suggest that the inflammation and oxidative stress induced by smoking lead to proteolytic imbalance and progressive lung structural derangement, with disease susceptibility being controlled by inherited variations in protective or inflammatory genes. However, cigarette smoke is directly mutagenic. Acquired somatic mutations, rather than inherited polymorphisms, might therefore be major determinants of COPD. Somatic mutations in oncogenes such as p53, Ras, EGFR and PTEN abound in the epithelium of smokers. These mutations are persistent, explaining the paradox that smoking cessation does not resolve inflammation. Moreover, the recognition that these somatic mutations converge on key inflammation, host defense and steroid response pathways might help to explain the clinical defects in these processes in COPD and guide discovery of future therapies. PMID- 12559771 TI - Obesity--a chronic health problem in cloned mice? AB - The majority of cloned animals derived by nuclear transfer die during gestation and display neonatal abnormalities. However, because of the long generation interval of cloned animal species, the long-term consequences of cloning on health have been difficult to access. Recent studies in mice have provided evidence that cloned animals might have chronic health problems such as obesity. Obesity in cloned mice is likely to reflect epigenetic abnormalities that arise partly from inadequate nuclear programming; these obese mice display a unique phenotype that is suggestive of a defect other than malfunction of the leptin melanocortin system, which occurs in most rodent models of obesity and in human obesity. PMID- 12559772 TI - Agonist induction and conformational selection during activation of a G-protein coupled receptor. AB - Substitutions of Asn111 of the AT(1) angiotensin receptor and mutations of the corresponding amino acids in other G-protein-coupled receptors (GPCRs) cause constitutive receptor activation. Ligand binding and signalling of constitutively active mutant GPCRs are discussed and similarities and differences during the activation of amine and peptide GPCRs are identified. Studies using the AT(1) receptor suggest that conformational selection is not sufficient to explain the mechanism of receptor activation, and that agonist binding to the receptor provides energy to induce activation of the receptor. Because agonist binding also actively facilitates the conformational rearrangements leading to activation of other GPCRs we propose that agonist induction should be considered as a general mechanism of GPCR activation. PMID- 12559773 TI - Principles: extending the utility of [35S]GTP gamma S binding assays. AB - Ligand regulation of the binding of [35S]GTPgammaS is one of the most widely used methods to measure receptor activation of heterotrimeric G proteins. However, until recently this method was largely restricted to receptors that interact with members of the family of pertussis-toxin-sensitive G proteins. Here, the reasons for this restriction are discussed and recent approaches that have extended the utility of this method such that it is now suitable for analysis of the activation of any heterotrimeric G protein are reviewed. PMID- 12559774 TI - Endothelial nitric oxide synthase: the Cinderella of inflammation? AB - A hallmark of inflammation is increased vascular permeability. Increases in vascular permeability and the migration of inflammatory cells are linked to complex interactions of inflammatory mediators with the vascular endothelium. Normally, endothelial nitric oxide synthase (eNOS) produces a tonic amount of nitric oxide (NO), which is responsible for the homeostasis between the endothelium and surrounding tissues. However, most agonists that act on endothelial cells cause a series of post-translational modifications that influence eNOS activity. Furthermore, stimulation by shear stress, autacoids or growth factors either induces eNOS or shifts it to a more active state, which produces a burst of NO. Here, we highlight recent findings about eNOS and propose how new pharmacological tools can be used to dissect the involvement and contribution of eNOS to inflammatory responses. PMID- 12559775 TI - Cyclooxygenase 2: a molecular target for cancer prevention and treatment. AB - Cyclooxygenase2 (COX-2), an inducible prostaglandin G/H synthase, is overexpressed in several human cancers. Here, the potential utility of selective COX-2 inhibitors in the prevention and treatment of cancer is considered. The mechanisms by which COX-2 levels increase in cancers, key data that indicate a causal link between increased COX-2 activity and tumorigenesis, and possible mechanisms of action of COX-2 are discussed. In a proof-of-principle clinical trial, treatment with the selective COX-2 inhibitor celecoxib reduced the number of colorectal polyps in patients with familial adenomatous polyposis. Selective COX-2 inhibitors appear to be sufficiently safe to permit large-scale clinical testing and numerous clinical trials are currently under way to determine whether selective inhibitors of COX-2 are effective in the prevention and treatment of cancer. PMID- 12559776 TI - Expression, polymorphism analysis, reticulocyte binding and serological reactivity of two Plasmodium vivax MSP-1 protein recombinant fragments. AB - Among the four parasite species causing malaria in humans, Plasmodium vivax prevails on both the Asian and the American continents. Several antigens from this parasite's erythrocytic stages have been characterised and some of them are considered to be good vaccine candidates. The P. vivax merozoite surface protein 1 (PvMSP-1) is a 200 kDa antigen, thought to mediate the initial contact between the merozoite and the erythrocyte. An effective blockage of this interaction could be important in anti-malarial vaccine design. This study analyses the genetic polymorphism, binding to both reticulocytes and erythrocytes, antigenicity and immunogenicity of two recombinant proteins belonging to the 33 kDa PvMSP-1 proteolytic fragment. Both regions showed very low genetic variation, bound reticulocytes with higher affinity than erythrocytes, were recognised by naturally P. vivax-infected patient sera and were immunogenic when used to immunise rabbits, making them good vaccine candidates against P. vivax, to be further preclinically tested in the Aotus monkey model. PMID- 12559777 TI - MMR vaccination advice over the Internet. AB - We wanted to investigate what advice UK homeopaths, chiropractors and general practitioners give on measles, mumps and rubella vaccination programme (MMR) vaccination via the Internet. Online referral directories listing e-mail addresses of UK homeopaths, chiropractors and general practitioners and private websites were visited. All addresses thus located received a letter of a (fictitious) patient asking for advice about the MMR vaccination. After sending a follow-up letter explaining the nature and aim of this project and offering the option of withdrawal, 26% of all respondents withdrew their answers. Homeopaths yielded a final response rate (53%, n = 77) compared to chiropractors (32%, n = 16). GPs unanimously refused to give advice over the Internet. No homeopath and only one chiropractor advised in favour of the MMR vaccination. Two homeopaths and three chiropractors indirectly advised in favour of MMR. More chiropractors than homeopaths displayed a positive attitude towards the MMR vaccination. Some complementary and alternative medicine (CAM) providers have a negative attitude towards immunisation and means of changing this should be considered. PMID- 12559778 TI - 'Clustering of exemptions' as a collective action threat to herd immunity. AB - In this paper, we examine the phenomenon of 'clustering of exemptions' to childhood vaccination, and the dangers this poses both to those exempted as well as the general population. We examine how clusters of exemptions might form through collective action as described by Thomas Schelling, and how religious groups who live in close proximity to one another can "self-select" in a way that exacerbates this phenomenon. Given the growing number of exemptions and the increasing visibility of the anti-vaccine movement, policy makers must be vigilant for dangerous clustering in order to avoid loss of herd immunity. PMID- 12559779 TI - Vaccination against type F botulinum toxin using attenuated Salmonella enterica var Typhimurium strains expressing the BoNT/F H(C) fragment. AB - The utility of the htrA, pagC and nirB promoters to direct the expression of the carboxy-terminal (H(C)) fragment of botulinum toxin F (FH(C)) in Salmonella enterica var Typhimurium has been evaluated. Only low levels of serum antibody were induced after immunisation, and some protection against botulinum toxin type F was demonstrated after oral immunisation of mice with two doses of any of these recombinant Salmonella. Immunisation with two doses of recombinant Salmonella expressing FH(C) from the htrA promoter gave the greatest protection, against up to 10,000 mouse lethal doses of botulinum toxin type F. These results demonstrate the feasibility of an orally delivered vaccine against botulinum toxin type F. PMID- 12559780 TI - Reduction of beta-amyloid plaques in brain of transgenic mouse model of Alzheimer's disease by EFRH-phage immunization. AB - Antibodies to the epitope EFRH, representing residues 3-6 within the beta-amyloid (Abeta) sequence, were previously shown to affect the solubility and disaggregation of Abeta fibrils in vitro. Here, we describe a novel method of immunization, using as antigen the EFRH peptide displayed on the surface of the filamentous phage. The EFRH phage evoked effective auto-immune antibodies in amyloid precursor protein [V717I] (APP[V717I]) transgenic mice that recapitulate the amyloid plaques and vascular pathology of Alzheimer's disease (AD). The immunization provoked a considerable reduction in the number of Abeta amyloid plaques in the brain of the transgenic mice and may serve as the basis for anti Abeta vaccine. PMID- 12559781 TI - HIV-specific immunity during structured antiviral drug treatment interruption. AB - The immunologic correlates associated with control of viremia in HIV disease are poorly understood. We hypothesized that structured antiviral drug treatment interruptions could be utilized to better understand the relationship between HIV specific immunity and viral replication. We thus examined the effects of two 8 weeks antiviral structured treatment interruptions (STIs) in a cohort of HIV-1 chronically infected individuals on highly active antiretroviral treatment (HAART) with (n = 13) and without (n = 12) therapeutic HIV immunizations. In this study, we observed that p24 gag antigen (np24) stimulated MIP-1beta levels and T helper immune responses prior to antiviral drug discontinuation were associated with control of viremia. Stronger and earlier production of gag peptide stimulated gamma interferon was observed in the immunized group during the structured antiviral drug interruptions. These results support the concept that HIV-specific immune responses are associated with control of viremia. Further study of immune-based therapies that enhance HIV-specific immunity is warranted. PMID- 12559782 TI - The out of Africa model of varicella-zoster virus evolution: single nucleotide polymorphisms and private alleles distinguish Asian clades from European/North American clades. AB - Until 1998, varicella-zoster virus (VZV) was generally considered sufficiently stable to allow the use of a single sequenced virus (VZV-Dumas) as a consensual representation of the world VZV genotype. But recent investigations have uncovered a gE mutant virus called VZV-MSP with a second genotype and a distinguishable accelerated cell spread phenotype. A subsequent study suggested that single nucleotide polymorphisms (SNPs) could be applied toward the genetic analysis of the VZV genome. To further assess the scope of genetic variation in the VZV genome on a worldwide basis, we carried out an extensive SNP analysis of structural glycoprotein genes gB, gE, gH, gI, gL, as well as the IE62 regulatory gene in viruses collected from Western Europe, North America and Asia, including the VZV vaccine strain. The SNP data showed segregation of viral isolates of Asian origin from those of Western ancestry into distinct phylogenetic clades. Unexpectedly, however, VZV from Thailand segregated with VZV from Iceland and the United States, i.e. it was more Western than Asian in nature. Further, SNP analysis disclosed strikingly unusual genotypes, e.g. gH genes with up to five missense mutations and gL genes with insertions of an in-frame methionine codon. In summary, these VZV genomic analyses have shown that individual VZV strains, like closely related human beings, have distinctive SNP profiles containing private alleles within just five VZV genes (gB, gH, gE, gL and IE62) that provide a fingerprint to localize ancestry of the viral strain. PMID- 12559783 TI - Eradication of established HPV16-transformed tumours after immunisation with recombinant Semliki Forest virus expressing a fusion protein of E6 and E7. AB - Previously, we described the efficacy of immunisation with recombinant Semliki Forest virus (SFV), expressing the human papillomavirus 16 (HPV) oncoproteins E6 and E7, in inducing HPV-specific CTLs and anti-tumour responses. Recently, we developed a novel recombinant SFV construct encoding a relatively stable fusion protein of HPV16 E6 and E7 under control of a translational enhancer derived from the SFV capsid protein. In the present study we demonstrate that immunisation of tumour-bearing mice with this improved vector results in the regression and complete elimination of established tumours. We furthermore demonstrate that a long-term high level of CTL activity, up to 340 days, accompanies the anti-tumour response. Thus, immunisation with recombinant SFV particles encoding increased levels of a fusion protein of HPV16 E6 and E7 efficiently induces CTL activity and CTL memory resulting in a potent therapeutic anti-tumour effect. PMID- 12559784 TI - DNA immunization with pgp3 gene of Chlamydia trachomatis inhibits the spread of chlamydial infection from the lower to the upper genital tract in C3H/HeN mice. AB - Chlamydia trachomatis pgp3 DNA immunized (no. 300) and non-immunized (no. 300) C3H/HeN mice were infected by vaginal inoculation with infectious C. trachomatis serotype D elementary bodies (EBs) and the spread of infection to the salpinges was assessed by cell culture isolation from tissue homogenates 7, 14, 21, 28, 35 and 42 days post-infection (p.i.). Overall, the pgp3-DNA immunization prevented salpinx infection in 94 (56%) mice, if compared with the 168 positive animals found among the non-immunized animals (P < 0.001). A group of negative control animals (i.e. mice immunized with plasmid DNA containing an irrelevant insert) was not protected, whereas all the mice of a positive immune control group (mice that had resolved a primary genital C. trachomatis infection) were resistant to re-infection. Pgp3 DNA immunization induced both humoral and mucosal anti-pgp3 antibodies. PMID- 12559785 TI - Seroprevalence of meningococcal serogroup C bactericidal antibody in England and Wales in the pre-vaccination era. AB - Sera from an age-stratified sample of 1689 individuals, submitted to the PHLS Seroepidemiology Unit between 1996 and 1999 were tested for serum bactericidal antibodies to serogroup C meningococci. Titres decreased during infancy, presumably as maternal antibody waned, and increased in older teenagers, the peak age of meningococcal carriage. The prevalence of antibody titres greater than or equal to 8 was highest in adults, with an average of 25% of adults 25 years old or above with titres above this putative protective level. In the absence of vaccination, antibody may be generated from periods of carriage of serogroup C meningococci, from other meningococcal strains sharing non-capsular antigens, and other cross-reactive organisms. The inverse relationship between disease incidence and the prevalence of 'protective' antibody titres as described by Goldschneider et al. appears more consistent with a titre of > or =8 rather than > or =128, although the proportions 'protected' are much lower here than in Goldschneider's study. This study provides baseline antibody levels which will facilitate the evaluation of the meningococcal serogroup C conjugate vaccination programme. PMID- 12559786 TI - Vaccination of puppies with a lipid-formulated plasmid vaccine protects against a severe canine distemper virus challenge. AB - We assessed whether the formulation of a DNA vaccine expressing the canine distemper virus (CDV) hemagglutinin (HA) and fusion (F) immunogens with the cationic lipid DMRIE-DOPE could induce serological responses and protection against a severe CDV challenge in the dog. Although clear protection was observed in dogs vaccinated with formulated plasmids only limited CDV specific antibody titers were observed in protected dogs before challenge, suggesting that protection could be explained by cell-mediated immunity and/or by a strong antibody-based memory response (priming) triggered by the infectious challenge. The high level of protection achieved in this study, demonstrated that formulated DNA CDV vaccines can generate in dogs a level a protection comparable to conventional CDV vaccines. PMID- 12559787 TI - Immunization with low doses of HIV-1 tat DNA delivered by novel cationic block copolymers induces CTL responses against Tat. AB - Cytotoxic T cell responses are key to the control of intracellular pathogens including HIV-1. In particular, HIV-1 vaccines based on regulatory proteins, such as Tat, are aimed at controlling HIV-1 replication and at blocking disease development by inducing cytotoxic T cell responses. Naked DNA is capable of inducing such responses but it requires several inoculations of high amounts of DNA, and/or prime-boost regimens. Here, we show that a novel class of cationic block copolymers protect the DNA from DNAse I digestion, and improve DNA delivery to antigen-presenting cells (APCs) after intramuscular (i.m.) vaccination. In particular, three cationic block copolymers (K1, K2 and K5) were used to deliver the HIV-1 pCV-tat DNA vaccine in BALB/c mice. The results indicate that vaccination with a very low dose (1 microg) of pCV-tat delivered by the cationic block copolymer K2 is safe and, as compared to naked DNA (up to 30 microg), greatly increases the CTL response against Tat, which was detected in all animals in the absence or in the presence of re-stimulation. PMID- 12559788 TI - Construction of designer glycoconjugate vaccines with size-specific oligosaccharide antigens and site-controlled coupling. AB - Coupling of carbohydrate antigens to protein carriers is a typical approach to enhancing the immunogenicity of carbohydrate-based vaccines. Glycoconjugates with well-defined structures are needed for studies defining the structural variables that govern antibody responses. We report a chemical strategy for preparation of an array of glycoconjugates containing saccharides of desired molecular sizes by selective depolymerization of bacterial polysaccharides and chemically controlled site-specific coupling. As an example, we synthesized and evaluated an oligosaccharide-based vaccine against type III group B Streptococcus. PMID- 12559789 TI - Total genome polymorphism and low frequency of intra-genomic variation in the uspA1 and uspA2 genes of Moraxella catarrhalis in otitis prone and non-prone children up to 2 years of age. Consequences for vaccine design? AB - Intra-genomic variation in the uspA1 and uspA2 genes of Moraxella catarrhalis was studied using pulsed field gel electrophoresis (PFGE) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. From a set of 91 M. catarrhalis isolates, 19 pairs of PFGE identical isolates were found. Five pairs originated from otitis non-prone children, 11 pairs from otitis prone children and for 3 pairs, one of the pair originated from an otitis prone and the other from an otitis non-prone child. No particular M. catarrhalis isolate was associated with either the otitis prone or non-prone children. One of these 19 pairs of isolates was found to exhibit both uspA1 and uspA2 intra-genomic variation, whilst another pair exhibited uspA2 intra-genomic variation only. Sequence data obtained from these variants showed that PCR-RFLP pattern differences reflected actual changes in predicted amino acid composition and that minor amino acid changes in a 23 base pair "NINNIY" repeat region (a conserved UspA1 and UspA2 binding site for the neutralising antibody mAb17C7) occurred. Variation in the uspA2 5' non-coding "AGAT" repeat region was also observed. These results may have implications for future M. catarrhalis vaccines comprising UspA1 or UspA2 components. PMID- 12559791 TI - Influenza vaccine uptake in the elderly: results from a rapid assessment of the effectiveness of new government policy in England for the winters 2000/2001 and 2001/2002. AB - Immunisation against influenza is an important means of reducing morbidity and mortality amongst high-risk groups, and especially the elderly. Although immunisation has been recommended for these groups for many years, no timely or comprehensive monitoring at the national level was carried out in England before the beginning of this century. Annual numbers of doses administered were available, but not to whom they were given. This paper describes the results of a national rapid reporting scheme for England that was introduced by the Department of Health (DH) and PHLS Communicable Disease Surveillance Centre (CDSC). The scheme collected monthly data on influenza immunisation uptake among people aged 65 or more during the winters of 2000/2001 and 2001/2002. Results showed that DH annual targets of 60 and 65%, respectively were both met, although considerable variation occurred at the local levels. The feasibility of this method of monitoring immunisation uptake in the elderly is discussed. PMID- 12559790 TI - Immunisation with modified HPV16 E7 genes against mouse oncogenic TC-1 cell sublines with downregulated expression of MHC class I molecules. AB - Human papillomavirus type 16 (HPV16)-transformed mouse TC-1 cells are extensively used in the evaluation of efficacy of experimental vaccines against tumours induced by HPVs. As these cells strongly express MHC class I molecules and downregulation of MHC class I surface expression is one of the important mechanisms that enable tumour escape from the host immune system, we undertook to derive TC-1 clones with reduced expression of MHC class I antigens. TC-1 cells were inoculated into mice preimmunised with an E7 gene-based DNA vaccine and from tumours developing in a portion of the animals, cell clones with downregulated MHC class I surface expression were isolated. Treatment with IFN-gamma resulted in an upregulation of MHC class I molecules in these cells, but after IFN-gamma removal, their expression gradually dropped again. When the expression of some components of the antigen-processing machinery (APM; LMP-2, TAP-1, and TAP-2) was tested, a reduced TAP-1 production was detected in cell lines with downregulated MHC class I expression. An enhanced immunoresistance of TC-1-derived clones with reduced MHC class I expression was observed in animals immunised with plasmids carrying modified E7 genes. Apart from the previously described fusion gene Sig/E7/LAMP-1, a new construct, Sig/E7GGG/LAMP-1, with a mutated Rb-binding site, was also used for immunisation. No significant change of immunogenicity was recorded for Sig/E7GGG/LAMP-1. Cell lines with downregulated MHC class I expression derived from TC-1 cells may represent a useful model for testing therapeutic anti-HPV vaccines in settings more relevant to clinical requirements. PMID- 12559792 TI - Ginseng and aluminium hydroxide act synergistically as vaccine adjuvants. AB - The dry extract prepared from the Panax ginseng C.A. Meyer-root (total ginseng (T ginseng)) contain ginsenosides (G-des) which were shown to have adjuvant properties as demonstrated by: (a) injecting guinea pigs with a mixture of T ginseng and inactivated porcine parvovirus (PPV) as a conventional vaccine; (b)injecting PPV-antigen and T-ginseng simultaneously but separately at different sites on the animal and (c)injecting only the T-ginseng 1 or 2 weeks prior to immunisation with the PPV-antigen. Using a haemagglutination inhibition (HI) test in the antibody titration, it was found that the mean HI-titre for the animals injected with PPV-antigen only was 320 +/- 0. By comparison, the mean titre value was 2026 +/- 1206 for the sera from the animals injected with the same vaccine but adjuvanted with 4 mg of T-ginseng, while the antibody titre induced by a vaccine containing Al(OH)(3)-gel was 2986 +/- 1596. Interestingly, the T-ginseng and Al(OH)(3) acted synergistically and further improved the antibody response to the PPV-antigen to 6826 +/- 2413, i.e. more than 20 times the HI titre of the non adjuvanted PPV-vaccine. Immunisations using PPV-vaccines adjuvanted with single purified G-des demonstrated that the ginseng fractions Rb1 and Rg1 are potent adjuvants inducing higher or similar antibody titres than the vaccine adjuvanted with Al(OH)(3), e.g. Rb1 tested at a concentration of 830 microg per dose induced a significantly (P = 0.009) higher antibody titre than the one adjuvanted with Al(OH)(3). Nevertheless, different than the mixture Al(OH)(3)-T-ginseng; Rb1 and Rg1 act antagonistically and partially inhibit each other. The G-des adjuvanted vaccines induced significantly (P = 0.0011) higher titres of IgG2 antibodies compared with IgG1. PMID- 12559793 TI - Rapid onset of protection following vaccination of calves with multivalent vaccines containing modified-live or modified-live and killed BHV-1 is associated with virus-specific interferon gamma production. AB - The objective of this study was to determine the effect of vaccination with commercially-available multivalent vaccines containing either modified-live (MLV) bovine herpesvirus-1 (BHV-1) (Bovishield) or MLV plus killed (MLV + K) BHV-1 (Reliant Plus) on protection against challenge at 5 days after a single vaccination. An additional objective was to determine whether cell-mediated immunity as measured by virus-specific interferon gamma (IFN-gamma) production by peripheral blood mononuclear cells (PBMC) was associated with any early protection induced by vaccination. Clinical signs, serum neutralizing (SN) titers, and nasal virus isolation (VI) titers were also measured. The 12-16-week old dairy cross-calves seronegative for antibodies to BHV-1 were vaccinated with a multivalent vaccine containing MLV BHV-1 (n = 19), a multivalent vaccine containing MLV + K BHV-1 (n = 19), or a control multivalent vaccine not containing BHV-1 (n = 10) on day 0 and challenged intranasally on day 5. PBMC were isolated on days 0, 3, 5, 8, 10, 14 and 19. PBMC were incubated in vitro with spent media, live BHV-1, or heat-inactivated BHV-1 for 72 h. Supernatants were assayed for bovine IFN-gamma by ELISA. Bovine herpesvirus-1-specific IFN gamma production was expressed as percent of the kit positive control, with value for spent media subtracted. Clinical signs were monitored daily. Serum VN titers were measured on days 0-5 and 19. Nasal VI titer was measured every other day from days 5 to 19. Interferon gamma production was higher on day 5, and was significantly increased post-challenge, in both vaccine groups compared to controls. There was no difference between vaccine groups on any day. There was no significant difference in SN titer among groups on any day. Virus isolation titer was significantly higher in controls on days 6 and 8 compared to both vaccine groups. Temperatures were significantly higher and nasal discharge was present more often post-challenge in controls compared to vaccine groups. Vaccination 5 days prior to challenge with commercially-available vaccine containing MLV or MLV + K BHV-1 was associated with increased BHV-1-specific IFN-gamma production, decreased viral shedding, lower temperatures and less nasal discharge post challenge. Cell mediated immune responses as measured by IFN-gamma production are stimulated rapidly following BHV-1 vaccination of calves. PMID- 12559794 TI - Mucosal and systemic antibody responses against an acellular pertussis vaccine in mice after intranasal co-administration with recombinant cholera toxin B subunit as an adjuvant. AB - To investigate the possibility of intranasal immunization with an acellular pertussis vaccine, groups of mice were administered intranasally with aluminium non-adsorbed pertussis toxoid (PTd; 0.5 or 5 microg) and formalin-treated filamentous hemagglutinin (fFHA; 5 microg) with and without recombinant cholera toxin B subunit (rCTB; 10 microg) as a mucosal adjuvant. At a low concentration of PTd, the following things became clear: (1) earlier and higher elevation of serum anti-PTd and anti-FHA IgG antibody titres in the presence of rCTB than in its absence, (2) higher serum anti-PTd and anti-FHA IgG antibody titres than 200 and 100 ELISA units ml(-1) (EU ml(-1)) in all mice, respectively, in the presence of rCTB, which were obtained by calibration against a reference anti-pertussis mouse serum, and (3) in an intranasal challenge experiment with Bordetella pertussis, slightly more rapid elimination of the bacteria from the lungs of mice intranasally immunized in the presence of rCTB, suggesting the effectiveness of rCTB as a mucosal adjuvant. However, irrespective of rCTB and dose of PTd, mice which were immunized four times and sacrificed on day 35 developed high levels of anti-PTd serum IgG antibodies, high or moderate levels of anti-FHA serum IgG antibodies and mucosal anti-PTd IgA antibodies in the lungs; only a slight or no increase of anti-FHA mucosal IgA antibodies was observed in the lung. These facts suggested the immunogenicity and mucosal adjuvanticity of PTd, and therefore, the mucosal adjuvanticity of rCTB seemed to be inconspicuous. Moreover, the addition of rCTB induced higher anti-PTd serum IgE antibody responses than no addition of it depending on dose of PTd. These results show that dose of PTd included in an acellular pertussis vaccine had better be low as possible and the addition of rCTB may not be always necessary in case of this nasal vaccine alone unlike tetanus and diphtheria toxoids and hepatitis B virus vaccine reported before. PMID- 12559795 TI - Revaccination of healthy nonresponders with hepatitis B vaccine and prediction of seroprotection response. AB - Sixty healthy nonresponders were randomized to receive intramuscular (IM) high dose hepatitis B virus (HBV) vaccine versus IM standard dose HBV vaccine plus granulocyte-macrophage colony-stimulating factor (GM-CSF) at 0-2 months. Antibody to hepatitis B surface antigen was measured 1 month after each dose and 3 months after the last dose. Two regimens were equivalent in eliciting seroprotection in nonresponders. Weight-height index (WHI) <39 and alanine transaminase (ALT) <39 mg/dl predicted which nonresponders would seroconvert. A three-dose regimen of standard dose HBV vaccine plus GM-CSF may be a useful for seroprotection of healthy nonresponders. PMID- 12559796 TI - Local and systemic immune response in nursing-home elderly following intranasal or intramuscular immunization with inactivated influenza vaccine. AB - Intramuscular (IM) influenza vaccines are only 30-40% effective in preventing clinical illness among the elderly, and their effectiveness in eliciting mucosal response may be even lower. The aim of the present study was to evaluate the immunological effect of a novel inactivated intranasal (IN) trivalent whole influenza virus vaccine among nursing-home elderly. Twenty-one institutionalized elderly subjects were vaccinated IN with an inactivated novel vaccine, twice, 21 days apart, and with no adverse effects. Twenty-two subjects were vaccinated once with a commercial IM vaccine. Viral strains used in the 1998/9 vaccine (20 microg of each per dose) were A/Beijing/262/95, A/Sydney/5/97 and B/Harbin/7/94. Serum antibodies (IgG and IgM) and nasal IgA were determined by the hemagglutination inhibition (HI) test and enzyme-linked immunosorbent assay (ELISA), respectively. Mucosal antibody response to the three vaccine strains was detected in 47.6-71.4% and 18.1-31.8% of IN and IM immunized subjects, respectively. Serum antibody response to the three antigens tested was detected in 20.0-61.9% and 18.2-72.7% of IN and IM immunized subjects, respectively. Seroconversion was not significantly different after IN or IM vaccination for both A/Sydney and B/Harbin, but higher for A/Beijing following IM vaccination. On study completion, 57.1, 65.0 and 50.0% of IN vaccinees were seroprotected to A/Beijing, A/Sydney and B/Harbin, respectively. Similarly, 68.1, 77.2 and 54.5% were immune after IM vaccination. The IN vaccine tested was significantly more effective than the IM vaccine in inducing mucosal IgA response. This may prevent influenza at its early stages and thus contribute to the reduction of morbidity and complications in nursing-home elderly. PMID- 12559797 TI - Enhancing the immunogenicity of bioengineered Listeria monocytogenes by passaging through live animal hosts. AB - Bioengineered Listeria monocytogenes can be used as a recombinant bacterial vaccine vector for the induction of strong cell-mediated immunity to passenger antigens. Listeria loses virulence after undergoing bioengineering techniques, thus decreasing its efficacy as a vaccine vector. We addressed this problem by examining the virulence, and the ability to induce CD8(+) T-cells, of Listeria monocytogenes vaccine strains before and after passaging through mice. We found that two in vivo passages are required to restore the induction of cell-mediated immunity to passenger antigens and maximum virulence to these strains. In addition, we found that after each passage, harvested bacteria must be cloned and checked for expression of the bioengineered gene to counter selection in favor of antigen loss mutants. PMID- 12559798 TI - Expression and immunogenicity of the major house dust mite allergen Der p 1 following DNA immunization. AB - The mite protein Der p 1 is a major trigger of allergy and atopic asthma world wide, and thus, a good vaccine candidate for allergy prevention. Since it is a cysteine protease, the catalytic effects of Der p 1 vaccination may be unpredictable. One approach to reduce this risk is to vaccinate with DNA encoding enzymatically inactive forms of Der p 1. Here we show that Der p 1 DNA without its native pre-pro sequences potently induced Der p 1-specific antibodies, as long as its pre-sequence was substituted by another leader sequence. Without any pre-pro sequence, the same DNA fragment was well expressed but failed to induce significant level of anti-Der p 1 antibodies, without further boosting by protein. PMID- 12559799 TI - Immunity to East Coast fever in cattle induced by a polypeptide fragment of the major surface coat protein of Theileria parva sporozoites. AB - Full-length recombinant versions of p67, the 709 amino acid major surface protein of Theileria parva sporozoites, induce immunity to East Coast fever (ECF) in cattle. We show that a soluble Escherichia coli recombinant version of p67 (p67(635)), in which a prokaryotic signal peptide replaces the eukaryotic one, confers protection comparable to that induced by the full-length molecule, but is unstable. Peptides encoding 80 (p67C) and 205 (p67N) amino acid fragments of p67, containing epitopes recognised by sporozoite neutralising monoclonal antibodies, exhibit improved stability in E. coli. Antibodies raised against the central region of p67 (p67M) neutralise sporozoite infectivity in vitro. The p67C peptide induced immunity against ECF in cattle, at a level equivalent to p67(635), suggesting that a synthetic peptide vaccine might be achievable. PMID- 12559800 TI - Carrier-dependent specificity of antibodies to a conserved peptide determinant of gp120. AB - Amino acid residues 421-436 constitute a comparatively conserved determinant of gp120 that participates in the binding of host cell CD4 receptors by HIV-1. We compared the immunogenicity of synthetic Cys-gp120 (421-436) conjugated to KLH via the N terminal Cys residue (KLH-I) and gp120 (421-436) extended at its N terminus with a 15 residue tetanus toxoid T cell epitope (T-I) in non-autoimmune mice (BALB/cstrain) and Fas-defective autoimmune mice (MRL/lpr strain). Both immunogens elicited high titer Abs detected as the binding to gp120 (421-436) conjugated to bovine serum albumin (BSA-I) immobilized in ELISA plates. Abs from KLH-I immunized mice displayed binding to full-length gp120 but the Abs from T-I immunized mice did not. Proteins unrelated in sequence to gp120 did not bind the Abs. Soluble I and T-I failed to compete with immobilized BSA-I for binding to anti-KLH-I Abs, whereas these peptides inhibited anti-T-I Ab binding by BSA-I (rank potency order: BSA-I > T-I >> I). These results indicate the influence of the carrier protein on the specificity of Abs to synthetic I. Low level BSA-I and gp120 binding Abs were detected in sera from non-immunized MRL/lpr mice. Similar Ab binding titers and specificity profiles were evident in MRL/lpr and BALB/c mice following immunization with KLH-I and T-I, indicating that pre-existing immunity to gp120 in the former strain does not influence the magnitude or specificity of the Ab response. PMID- 12559801 TI - Relationship between the degree of antigen adsorption to aluminum hydroxide adjuvant in interstitial fluid and antibody production. AB - The effect of the degree of adsorption after exposure to interstitial fluid on the immune response in mice to model vaccines containing ovalbumin, alpha casein or dephosphorylated alpha casein adsorbed to aluminum hydroxide adjuvant was studied. Ovalbumin and dephosphorylated alpha casein were adsorbed in the vaccine but were completely eluted when exposed to interstitial fluid for 4 h. The presence of aluminum hydroxide adjuvant in the vaccine produced immunopotentiation compared to a solution of the protein even though the protein desorbed rapidly upon subcutaneous administration. In contrast, alpha casein was completely adsorbed to aluminum hydroxide adjuvant in both the vaccine and upon exposure to interstitial fluid. Immunopotentiation by aluminum hydroxide adjuvant was also observed in this model vaccine compared to a solution of alpha casein. The results indicated that antigen presenting cells can take up desorbed antigen from interstitial fluid as well as antigen adsorbed to aluminum-containing adjuvants. PMID- 12559802 TI - Safety and immunogenicity of a live-attenuated influenza vaccine blended and filled at two manufacturing facilities. AB - This study was designed to compare the safety and immunogenicity of a trivalent live-attenuated, cold-adapted influenza vaccine (CAIV-T) blended and filled at two different manufacturing facilities (Medeva and Aviron-PA). The vaccines contained approximately 10(7) TCID(50) (median tissue culture infectious dose) of each of the three recommended 1997-1998 influenza vaccine components, A/Shenzhen/227/95 (H1N1) (A/Bayern/7/95 (H1N1)-like strain), A/Wuhan/359/95 (H3N2), and B/Ann Arbor/1/94 (B/Beijing/184/93-like strain). Two hundred and twenty-five healthy Australian children aged 12-42 months were enrolled and randomized in a 3:2 ratio to receive CAIV-T blended and filled either at Medeva or at Aviron-PA. Two doses of CAIV-T were given 4-6 weeks apart as an intranasal spray. Three blood specimens were collected (immediately before doses one and two, and 28 +/- 5 days following dose two) for measuring hemagglutination inhibition (HAI) antibody responses. Adverse events occurring within 10 days and serious adverse events occurring within 42 days were collected. Serum HAI antibody levels were measured against the three vaccine strains. Equivalent immunogenicity between the two vaccine groups was pre-specified as: (1) within 20% difference in seroconversion rates (HAI titers > or =4-fold rise); and (2) within 4-fold difference in the 90% confidence interval of geometric mean titer ratio. Among 10 pre-specified adverse events, only vomiting had significantly different incidence rates in the two vaccine groups following dose one (3% versus 13%, P = 0.01) but the difference disappeared following dose two (4% versus 4%). Differences in seroconversion rates following dose two between the two vaccine groups in pre-vaccination seronegative children were all <20% for the three vaccine strains (16% for H1N1, 0% for H3N2, and 0% for B). The results indicate that CAIV-T blended and filled in the two facilities had equivalent profiles of safety and immunogenicity. PMID- 12559803 TI - Recombinant raccoon pox vaccine protects mice against lethal plague. AB - Using a raccoon poxvirus (RCN) expression system, we have developed new recombinant vaccines that can protect mice against lethal plague infection. We tested the effects of a translation enhancer (EMCV-IRES) in combination with a secretory (tPA) signal or secretory (tPA) and membrane anchoring (CHV-gG) signals on in vitro antigen expression of F1 antigen in tissue culture and the induction of antibody responses and protection against Yersinia pestis challenge in mice. The RCN vector successfully expressed the F1 protein of Y. pestis in vitro. In addition, the level of expression was increased by the insertion of the EMCV-IRES and combinations of this and the secretory signal or secretory and anchoring signals. These recombinant viruses generated protective immune responses that resulted in survival of 80% of vaccinated mice upon challenge with Y. pestis. Of the RCN-based vaccines we tested, the RCN-IRES-tPA-YpF1 recombinant construct was the most efficacious. Mice vaccinated with this construct withstood challenge with as many as 1.5 million colony forming units of Y. pestis (7.7 x 10(4)LD(50)). Interestingly, vaccination with F1 fused to the anchoring signal (RCN-IRES-tPA-YpF1-gG) elicited significant anti-F1 antibody titers, but failed to protect mice from plague challenge. Our studies demonstrate, in vitro and in vivo, the potential importance of the EMCV-IRES and secretory signals in vaccine design. These molecular tools provide a new approach for improving the efficacy of vaccines. In addition, these novel recombinant vaccines could have human, veterinary, and wildlife applications in the prevention of plague. PMID- 12559804 TI - Co-immunization with plasmids coding the full length and a soluble form of glycoprotein D of HSV-2 induces protective cellular and humoral immune response in mice. AB - At present, the significance of antibody for protection of the female genital tract against infection with HSV-2 remains controversial. In the present study, the ability of a DNA vaccine encoding different forms of glycoprotein D (gD) of herpes simplex virus-2 (HSV-2) to induce simultaneously cellular and humoral responses was evaluated. Mice immunized with a plasmid encoding full length gD (pgD) developed a strong cellular immune response but weak antibody titers in serum and vaginal washings. On the other hand, mice immunized with a plasmid encoding soluble form of gD (pdeltagD) showed high titers of antibodies but a very weak cell-mediated immune response. When mice were immunized simultaneously with both plasmids, cellular and humoral immune responses were elicited. This mice showed neutralizing antibodies in serum and vaginal washings as well as a high number of IFN-gamma secreting cells in spleen. When challenged with 50 lethal doses of virus, mice immunized with pgD along with pdeltagD showed a more complete protection than mice immunized with pgD alone. Collectively these results suggest that neutralizing antibodies help cell-mediated immune response for the protection against HSV-2 infection. PMID- 12559805 TI - Factors affecting the compliance of the antenatal hepatitis B screening programme in Italy. AB - The effectiveness in the prevention of perinatally transmitted HBV infection was assessed in 11858 pregnant women consecutively recruited in public and private hospitals in six Italian regions during a 2 months period in 2001. Of them 10881 (91.8%) attended HBsAg antenatal screening. The overall HBsAg prevalence was 1.7% (CI 95%: 1.4-1.9); it was 1.4% (CI 95%: 1.2-1.7) in pregnant women born in Italy but 5.9% (CI 95%: 4.1-8.1) in those born in Asia, Africa, central and south America, and eastern Europe. Results of multiple logistic regression analysis indicate that birth in foreign countries (OR 2.0; CI 95%: 1.3-3.0), family size with more than 4 members in the household (OR 3.5; CI 95%:2.7-4.6), and birth in a private hospital (OR 1.9; CI 95%: 1.3-2.8) were all independent predictors of lack of adherence to HBsAg screening. Out of the 182 new-borns of HBsAg positive mothers 172 (95.0%) were given active plus passive immunisation; this figure was 100% in new-borns of foreign mothers. These findings evidence a good effectiveness in the prevention of perinatally transmitted HBV in Italy. More efforts should be addressed to improve the effectiveness of the programme among foreign pregnant women who have high rate of HBsAg and more likely escape HBsAg screening than Italian pregnant women. PMID- 12559806 TI - Encapsulation of peptides in biodegradable microspheres prolongs their MHC class I presentation by dendritic cells and macrophages in vitro. AB - Biodegradable microspheres (MS) consisting of poly(D,L-lactide-co-glycolide) (PLGA) represent a promising alternative to conventional adjuvants. The adjustable pulsatile release of encapsulated material from such MS offers the potential to mimic the priming and boosting injections of conventional immunization regimens. In this paper, we demonstrate that MS can serve as antigen reservoirs in antigen presenting cells (APC), so that antigen is presented for extended periods of time (up to 9 days). In particular, we could show by measurement of IFN-gamma production that encapsulated peptides were presented to cytotoxic T lymphocytes (CTL) by mouse and human macrophages as well as by human dendritic cells in vitro for a longer time period as compared to soluble peptides. The extended antigen presentation may thus improve the CTL response in vivo. These results may be of paramount importance in cancer vaccination therapy since MS may serve as antigen reservoirs to extend the presentation time by APC used to boost the patient's immune response to tumor antigens. PMID- 12559807 TI - The cost-effectiveness of routine childhood varicella vaccination in Germany. AB - This study explores the economic value of a routine varicella vaccination program for Germany. An age-structured decision analytic model was used to assess the benefits, costs and cost-effectiveness of an immunization program for a period of 30 years. Three interventions were compared with no vaccination: universal vaccination of around 15 months old healthy children, vaccination of susceptible adolescents (11-12 years of age), and the combined strategy. The analysis was conducted from both the societal perspective and the payers', i.e. sickness funds, perspective. Input data were mainly derived from a retrospective survey (analyzed were 1334 patient records) and from a seroprevalence study (n = 4602 sera). Using a coverage rate of 85% and a vaccine efficacy rate of 86% routine children vaccination could prevent around 611,000 varicella cases and over 4700 major complications per year. Average yearly cost savings for the society are 51.3 million Euro. The benefit-cost ratio (BCR) is 4.12. From the third-party payer's perspective, the BCR is 1.75 which is a consequence of significant reimbursement of parent's lost earnings by German sickness funds. The adolescent vaccination strategy has a favorable BCR ratio of 8.44 from the societal perspective, but clearly inferior medical effects. The combined vaccination strategy showed similar results as the children strategy. Routine childhood varicella vaccination appears to be a highly efficient strategy to reduce the burden of varicella and results in significant savings for both the society and the payers. PMID- 12559808 TI - Comparison of the reactogenicity and immunogenicity of a split and a subunit adjuvanted influenza vaccine in elderly subjects. AB - A randomised, open study was carried out among an elderly population in order to compare the reactogenicity and immunogenicity of an inactivated, split virion influenza vaccine (Vaxigrip, Aventis Pasteur MSD, Lyon, France) with that of an MF59-adjuvanted, subunit vaccine (Fluad, Chiron Vaccines, Siena, Italy). Both vaccines contained the three strains: A/Sydney/5/97 (H3N2), A/Beijing/262/95 (H1N1) and B/Beijing/184/93, recommended by the WHO for the 1998-1999 influenza season. A total of 2150 subjects were vaccinated and included in the reactogenicity analysis. A total of 1076 subjects received Vaxigrip (age 73.3 +/- 5.9 years, 49.6% men) and 1074 subjects received Fluad (age 73.4 +/- 5.9 years, 52.3% men). All subjects were kept under medical observation for 30 min after vaccination, in order to check any immediate local and/or systemic reaction. A self monitoring diary card was given to all subjects to collect any local and/or systemic reaction occurring during the 3 days following the vaccination, any adverse event occurring between vaccination day and 21st day post-vaccination and any medication taken during the study period. A total of 1186 subjects were included in the immunogenicity analysis. A total of 591 subjects received Vaxigrip (age 73.4 +/- 5.6 years, 52.3% men) and 595 subjects received Fluad (age 73.8 +/- 5.9 years, 55.8% men). Blood samples were collected pre- and 21 days post-vaccination and were analysed by the haemagglutination inhibition assay. In terms of reactogenicity both vaccines were generally well tolerated. The frequency of local reactions was lower in the group that received Vaxigrip. Pain at the injection site occurring from 30 min to 3 days after vaccination was also significantly less frequent (P = 0.005) in the Vaxigrip group. Fever > or =37.5 degrees C was reported in less than 1% of all vaccinated subjects. No serious adverse event was related to vaccine administration. In terms of immunogenicity both vaccines induced an effective immune response (anti-HI titre > or =40) against A/Sydney/5/97 (H3N2) and A/Beijing/262/95 (H1N1) strains in the entire population. Vaxigrip and Fluad induced similar seroprotection and seroconversion rates against the A/Sydney/5/97 (H3N2) strain. For both vaccines a lower percentage of subjects achieved a seroprotective titre > or =40 against the B/Beijing/184/93. A lower antibody response against the influenza B strain was also observed in other studies conducted during the same season. In subjects 75 years of age or older, Fluad was more immunogenic than Vaxigrip for all three virus strains. PMID- 12559809 TI - In ovo delivery of DNA to the avian embryo. AB - We have developed a simple method of transfecting avian embryos in ovo with various plasmid vectors that results in protein expression in the embryo. Using the chloramphenicol acetyl transferease (CAT) reporter gene, we were able to show that transfecting avian embryos with a plasmid/neutral lipid/dimethylsulfoxide (DMSO) mixture delivered to the air cell, is better than transfecting naked DNA or cationic lipid encapsulated DNA, using DMSO (P < 0.05). This method resulted in CAT expression in several avian embryonic tissues of all the embryos inoculated. We found that both the cytomegalovirus (CMV) and chicken beta actin promoters worked significantly better (P < 0.05) than the Rous sarcoma virus promoter in vitro for reporter gene expression after cationic liposome-mediated transfection. However, after in ovo delivery with neutral lipid encapsulation and DMSO mediated delivery, no significant difference (P > 0.05) between the various promoters could be determined. We believe this neutral lipid encapsulation method may represent an important platform for delivery of DNA to the avian embryo. PMID- 12559810 TI - Construction and characterisation of O139 cholera vaccine candidates. AB - The hemagglutinin/protease (HA/P) seems to be an attractive locus for the insertion of heterologous tags in live cholera vaccine strains. A deltaCTXphi spontaneous mutant derived from a pathogenic strain of O139 Vibrio cholerae was sequentially manipulated to obtain hapA Colon, two colons celA derivatives which were later improved in their environmental safety by means of a thyA mutation. All the strains here obtained showed similar phenotypes in traits known to be remarkable for live cholera vaccines irrespective of their motility phenotypes, although the hapA mutants had a 10-fold decrease in their colonisation capacity compared with their parental strains in the infant mouse cholera model. However, the subsequent thyA mutation did not affect their colonisation properties in the same model. These preliminary results pave the way for further clinical assays to confirm the possibilities of these vaccine prototypes as safe and effective tools for the prevention of O139 cholera. PMID- 12559813 TI - Investigation of nanocomposites based on semi-interpenetrating network of [L-poly (epsilon-caprolactone)]/[net-poly (epsilon-caprolactone)] and hydroxyapatite nanocrystals. AB - In this paper the semi-interpenetrating network (semi-IPN) technique was used for the first time to prepare bone implant composites containing hydroxyapatite (HAP) nanocrystals. The prepared nanocomposites are expected to combine several property advantages including good mechanical strength, modified degradation rate and excellent osteoconductivity. The semi-IPN matrix based on the linear poly (epsilon-caprolactone) (L-PCL) and the network poly (epsilon-caprolactone) (net PCL) structures are revealed to be phase separation structures. The morphology of net-PCL is featured by intracrosslinked microdomains (1-10 microm) that further interconnect with each other to form the network over the whole sample. The net PCL component is totally amorphous at room temperature for the nanocomposites containing HAP up to 12.3 wt%. Further, the crystallinity of L-PCL is greatly decreased due to the presence of net-PCL as compared with that for pure L-PCL. The incorporation of L-PCL into the net-PCL network could significantly improve the mechanical properties of pure net-PCL. A great improvement in mechanical properties is observed for the nanocomposites if the HAP content is increased to 15.8 wt%. This transition is in agreement with that the net-PCL component changes from amorphous state to crystalline state at this composition. PMID- 12559811 TI - Vaccination with DNA encoding ORFF antigen confers protective immunity in mice infected with Leishmania donovani. AB - The gene ORFF is part of the multigenic LD1 locus on chromosome 35 that is frequently amplified in Leishmania. The function of ORFF is unknown. The gene encoding ORFF was cloned into a eukaryotic expression vector downstream to the cytomegalovirus (CMV) promoter. BALB/c mice were injected intramuscularly with ORFF DNA and challenged with Leishmania donovani promastigotes. Vaccination with ORFF gene induced both humoral and cellular immune response against ORFF, which provided significant level of protection against challenge with L. donovani. A qualitative PCR was used to determine whether activation of Th1 cells develops selectively in response to this ORFF DNA vaccine. The results indicated that mRNA for IFN-gamma was significantly induced in immunized mice. No significant change in IL-4 mRNA expression was observed in mice immunized with ORFF DNA vaccine versus mice immunized with control plasmid. Thus, DNA immunization may offer an attractive alternative strategy against leishmaniasis. PMID- 12559814 TI - Inorganic plasma with physiological CO2/HCO3- buffer. AB - The substitution of tris/HCl buffer by CO(2)/HCO(3)(-) buffer in inorganic plasma was studied. An appropriate gas mixture of CO(2)/N(2) was continuously bubbled in Kokubo's SBF solution prepared without addition of Tris/HCl. This method enables buffering the solution within the 7.3-7.4 pH interval and, at the same time, reaching a HCO(3)(-) concentration between 24 and 27 mmol dm(-3), which are the normal concentrations reported for blood plasma. Mineralisation studies on calcium phosphate ceramics using this solution showed that, in the presence of such hydrogencarbonate concentrations, the formation of a mineralisation layer on the ceramic occurs via a carbonated octacalcium phosphate, that evolves to carbonated hydroxyapatite. The results suggest that mineralisation studies in this new carbonate-containing simulated inorganic plasma mimic biomineralisation more closely than traditional SBF. PMID- 12559815 TI - Comparison of the antioxidant properties of wound dressing materials- carboxymethylcellulose, hyaluronan benzyl ester and hyaluronan, towards polymorphonuclear leukocyte-derived reactive oxygen species. AB - In chronic wounds, factors are released which perpetuate inflammatory processes, including polymorphonuclear leukocyte (PMN)-derived reactive oxygen species (ROS), such as superoxide radical (O(2)*-) and hydroxyl radical (*OH) species. The glycosaminoglycan, hyaluronan, has established antioxidant properties towards ROS, although the antioxidant potential of wound dressing biomaterials, such as 75% benzyl esterified hyaluronan (BEHA) and carboxymethylcellulose (CMCH), are less characterised. This study compared the antioxidant properties of high and low molecular weight hyaluronan (HMWT HA and LMWT HA), BEHA and CMCH towards ROS, generated by stimulated PMN in vitro. The antioxidant capacities of each biomaterial were assessed by their inhibition of O(2)*- -induced cytochrome C reduction, generated via PMN stimulation by phorbol myristyl acetate (PMA); and their inhibition of *OH-induced 2-deoxy-D-ribose degradation, generated by PMA stimulated PMN in the presence of a ferric chloride-EDTA chelate. All biomaterials, except LMWT HA, possessed dose-dependent antioxidant properties against O(2)*-, BEHA having greatest antioxidant potential, followed by HMWT HA and CMCH. HMWT HA exhibited the highest dose-dependent antioxidant properties towards *OH, followed by BEHA and CMCH. LMWT HA demonstrated no antioxidant properties towards *OH. These antioxidant activities, particularly towards O(2)* , may be beneficial in removing the initial source of ROS necessary for the secondary formation of *OH, implicated as a causal factor for the extensive metabolic alterations observed in chronic wounds. PMID- 12559816 TI - Platelets stimulated by IgG-coated surfaces bind and activate neutrophils through a selectin-dependent pathway. AB - Blood platelets bind rapidly to foreign surfaces and interact with adsorbed proteins and neutrophil granulocytes. We demonstrate by use of luminol-amplified chemiluminescence under stirred and non-stirred conditions that platelets at IgG coated surfaces amplify the neutrophil extracellular release of reactive oxygen species (ROS). The neutrophil response involved tyrosine phosphorylation, but was only in part induced by neutrophil F(c gamma)-receptor stimulation. The platelet mediated effects were contact-dependent since the respiratory burst was inhibited when the IgG-stimulated platelets were removed by filtration, but not when they were fixed in paraformaldehyde. Bodipyphallacidin-staining of filamentous actin (F-actin) revealed that an actin-dependent platelet adhesion supported the subsequent adhesion and spreading of neutrophils. The neutrophil ROS-response was lowered when the interaction between platelet P-selectin (CD62P) and neutrophil P selectin glycoprotein ligand-l (PSGL-1 or CD162) was inhibited. The blocking of L selectin (CD62L) or blocking of the interaction between platelet glycoprotein (Gp) IIb/IIIa and neutrophil complement receptor 3 (CR3) showed no effect. We conclude that platelet activation on immobilized IgG trigger a contact-dependent "frustrated" phagocytosis by neutrophils, associated with a release of toxic ROS. PMID- 12559817 TI - Surface characterization of passive film on NiCr-based dental casting alloys. AB - Surface characterization of passive film formed on different NiCr-based dental casting alloys in an acidic artificial saliva was investigated by using electrochemical impedance spectroscopy (EIS) measurement and potential decay test. The polarization resistance (R(p)), which is inversely proportional to corrosion rate, of passive film was obtained from the EIS data. The stable passive current density, passivating rate, and passivity decay rate of passive film were evaluated by potential decay test. Surface chemical analyses of passive film were performed by using X-ray photoelectron spectrometer and Auger electron spectrometer. Results showed that the R(p), stable passive current density, passivating rate, and passivity decay rate, respectively, were significantly different among the various NiCr-based alloys (p<0.001). The presence of higher Cr (maximum 31-37 at%; mainly as Cr(2)O(3)) and Mo (maximum 10-11 at%; mainly as MoO(3)) contents in the passive film led to a higher R(p), lower stable passive current density, faster passivating rate, and lower passivity decay rate. The presence of Be (>20 at%; mainly as BeO) in the passive film was detrimental to both the corrosion resistance and the stability of passive film. The presence of small amount of Ti (<6 at%; mainly as TiO(2)) in the passive film did not show any influence on the characterization of passive film. PMID- 12559818 TI - Mechanical and histomorphometric evaluations of titanium implants with different surface treatments inserted in sheep cortical bone. AB - Improvement of the implant-bone interface is still an open problem and the interest in chemical modification of implant surfaces for cementless fixation has grown steadily over the past decade. Mechanical and histomorphometric investigations were performed at different times on implants inserted into sheep femoral cortical bone to compare the in vivo osseointegration of titanium screws ( X 3.5 x 7 mm length) with different surface treatments. After 8 weeks of implantation, the push-out force of anodized and hydrothermally treated implants (ANODIC) was significantly higher than that of machined implants (MACH) (36%, p<0.0005), whereas a decrease of 39% was observed for acid-etched implants (HF) when compared to other surface treatments. After 12 weeks of implantation, the push-out force values of HF implants were still significantly lower than those observed for MACH (-19%, p<0.01) and hydroxyapatite vacuum plasma-sprayed implants (HAVPS, -25%, p<0.0005), and the highest push-out force was found in HAVPS (p<0.001) implants. After 8 and 12 weeks of implantation, the AI of HF implants was significantly (p<0.05) lower ( approximately -25%) than that of MACH, HAVPS and ANODIC implants. In conclusion, results appear to confirm that there are no specific differences between ANODIC and HAVPS implants in terms of behavior. Moreover, although MACH implants show some surface contaminating agents, they appear to ensure good osseointegration within 12 weeks both mechanically and histomorphometrically, as do ANODIC and HAVPS implants. However, further studies are required to investigate bone hardness and mineralization around implants. PMID- 12559819 TI - Preparation of water-soluble chitosan/heparin complex and its application as wound healing accelerator. AB - To make effective wound healing accelerator, water-soluble chitosan (WSC)/heparin (CH) complex was prepared using WSC with wound healing ability and heparin with ability to attract or bind growth factor related to wound healing process. Water soluble CH complex was prepared by the reaction between WSC and heparin, and then, by adding distilled water to it, ointment type with high viscosity was made. To evaluate the wound healing effect, full thickness skin excision was performed on the backs of the rat and then WSC and water-soluble CH complex ointments were applied in the wounds, respectively. After 15 days, gross and histologic examination was performed. Grossly, untreated control group revealed that the wound had well defined margin and was covered by crust. The second group treated with WSC ointment revealed small wound size with less amount of covering crust and ill-defined margin, which appeared to regenerate from margin. The third group treated with water-soluble CH complex ointment appeared to be nearly completely healed. Histology of each group was well correlated to gross findings. The third group shows nearly complete regeneration of appendage structure similar to normal in the dermis in contrast to control and second group with absence and less number of skin appendages, respectively. PMID- 12559820 TI - In vitro studies of plasma-sprayed hydroxyapatite/Ti-6Al-4V composite coatings in simulated body fluid (SBF). AB - The bioactivity of plasma-sprayed hydroxyapatite (HA)/Ti-6Al-4V composite coatings was studied by soaking the coatings in simulated body fluid (SBF) for up to 8 weeks. This investigation was aimed at elucidating the biological behaviour of plasma-sprayed HA/Ti-6Al-4V composite coatings by analyzing the changes in chemistry, and crystallinity of the composite coating in a body-analogous solution. Phase composition, microstructure and calcium ion concentration were analyzed before, and after immersion. The mechanical properties, such as tensile bond strength, microhardness and Young's modulus were appropriately measured. Results demonstrated that the tensile bond strength of the composite coating was significantly higher than that of pure HA coatings even after soaking in the SBF solution over an 8-weeks period. Dissolution of Ca-P phases in SBF was evident after 24h of soaking, and, a layer of carbonate-apatite covered the coating surface after 2 weeks of immersion. The mechanical properties were found to diminish with soaking duration. However, slight variation in mechanical properties was found after supersaturation of the calcium ions was attained with the precipitation of the calcium phosphate layers. PMID- 12559821 TI - An initial investigation of photocurable three-dimensional lactic acid based scaffolds in a critical-sized cranial defect. AB - Degradable polymer networks formed by the photoinitiated polymerization of multifunctional monomers have great potential as in situ forming materials, especially for bone tissue engineering. In this study, one specific chemistry was analyzed with respect to bone formation in a critical-sized defect model with and without adsorbed osteoinductive growth factors present. The scaffolds degraded in approximately 8 months and possessed an elastic modulus similar to that of trabecular bone. A porous scaffold fabricated with approximately 80% porosity and pore diameters ranging from 45 to 150 mm was implanted in a critical-sized cranial defect in rats. When implanted alone, the scaffolds were filled primarily with fibrous tissue after 9 weeks with only mild inflammation at the defect site. When the scaffolds released osteoinductive growth factors, statistically more bone filled the scaffold. For instance, 65.8+/-9.4% (n=5) of the defects were filled with radiopaque tissue in the osteoinductive releasing scaffolds, whereas only 24.2+/-7.4% (n=5) of the defects were filled in the untreated defects 9 weeks after implantation. These results illustrate not only the benefits of delivering osteoinductive factors when developing synthetic bone grafts, but the potential of these materials for supporting the infiltration and development of bone in large defects. PMID- 12559823 TI - Biological characterization of EDC-crosslinked collagen-hyaluronic acid matrix in dermal tissue restoration. AB - Porous collagen matrices crosslinked with various amounts of hyaluronic acid (HA) by 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide (EDC) were developed as scaffolds for dermal tissue regeneration. The effect of HA on cells in accordance with HA concentrations in the collagenous matrices was investigated using cultures of fetal human dermal fibroblasts, and the effect of EDC-crosslinked collagen-HA matrix on wound size reduction was also evaluated in vivo. Scanning electron microscopic views of the matrices demonstrated that all of the collagen HA matrices had interconnected pores with mean diameters of 150-250 microm. An HA matrix retention test showed that the concentration of HA decreased slowly after an initial rapid decrease over 24h. Fetal human dermal fibroblasts adhered well to all of the collagen-based matrices as compared with the Porous polyurethane matrix used as a control. An 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide based proliferation test and the hematoxylin and eosin staining of a 2 week cultured matrix showed that the proliferation of fibroblasts was enhanced on a 9.6% HA contained collagen matrix. No significant difference was in terms of fibroblast migration into the various types of scaffolds as HA content was increased. In vivo testing showed that dermis treated with collagen or collagen HA matrix was thicker than the control, and epithelial regeneration was accelerated, and collagen synthesis increased. However, no significant effect of HA on wound size reduction was found. PMID- 12559822 TI - The properties of chitosan-gelatin membranes and scaffolds modified with hyaluronic acid by different methods. AB - The objective of the present study was to investigate the properties of chitosan gelatin membranes or scaffolds, which were modified by incorporation of hyaluronic acid in the surface or bulk phase through co-crosslinking with N,N-(3 dimethylamino-propyl)-N'-ethyl carbodiimide (EDC) and N-hydroxysuccinimide (NHS) in 2-morpholinoethane sulfonic acid (MES) buffer. The comparative study on properties of surface modification (HA(S)) and polyblend membranes (HA(C)) revealed that gelatin was enriched on the surface of HA(C), while hyaluronic acid was enriched on the surface of the HA(S). The HA(S) membranes made by surface modification method had a characteristic surface morphology. The corresponding scaffolds were prepared through freeze-drying. The incorporation of hyaluronic acid improved flexibility and fibroblasts adhesion, while slowing down the rate of biodegradation of chitosan-gelatin scaffold. Human fibroblasts adhered and proliferated well on the membranes or scaffolds in vitro. PMID- 12559824 TI - Bone regeneration by recombinant human bone morphogenetic protein-2 and a novel biodegradable carrier in a rabbit ulnar defect model. AB - The effects of recombinant human bone morphogenetic protein (rhBMP)-2 and a novel carrier, PLGA-coated gelatin sponge (PGS), on bone defect repair was examined. A 1.5 cm unilateral segmental bone defect was created in the ulnar diaphysis of a Japanese white rabbit. In an initial study, defects were either treated with PGS impregnated with various concentrations of rhBMP-2 (0, 0.1, 0.4 and 1 mg/cm(3)) or left untreated. Defect healing was assessed by radiographic union rate, and biomechanical properties of regenerated bones were determined at 16 weeks postoperatively. In a second study, defects were implanted with PGS with or without rhBMP-2, and histologically observed at postoperative weeks 8 and 16. Radiographic union rate increased the dose-dependently at an early time point. All defects treated with rhBMP-2 (0.4 and 1 mg/cm(3)) were radiographically repaired. Mechanical properties of regenerated bones were restored in a dose dependent manner. Neither ulnae left untreated nor implanted PGS alone showed radiographic union. Longitudinal alignment of lamellar structure was observed histologically at 16 weeks, indicating that remodeling of regenerated bone was complete. Implanted PGS was almost completely resorbed by 8 weeks, and no abnormalities were observed in the surrounding soft tissue. These results suggest that PGS is a promising carrier for rhBMP-2. PMID- 12559825 TI - Nerve regeneration in a collagen-chitosan tissue-engineered skin transplanted on nude mice. AB - A reconstructed skin made of a collagen-chitosan sponge seeded with human fibroblasts and keratinocytes and grown in vitro for 31 days was developed for the treatment of deep and extensive burns. The aim of this study was to assess whether this tissue-engineered skin could promote nerve regeneration in vivo, since recovery of sensation is a major concern for burnt patients. The human reconstructed skin was transplanted on the back of nude mice and the growth of nerve fibres within it was assessed 40, 60, 90 and 120 days after graft. Nerve growth was monitored by confocal microscopy using immunohistochemical staining of PGP 9.5 and 150 kD neurofilament, while Schwann cell migration was observed using protein S100 expression and laminin deposition. Nerve growth was first detected 60 days after transplantation and was more abundant 90 and 120 days after graft. Linear arrangements of Schwann cells were observed in the graft as early as 40 days after graft. Nerve growth was observed along these Schwann cell extensions 60 days after transplantation. We conclude that the three-dimensional architecture of the collagen-chitosan tissue-engineered skin sponge encourages nerve growth. This result provides new perspectives to increase nerve regeneration within the tissue-engineered skin by linkage of neurotrophic factors in the sponge before transplantation. PMID- 12559826 TI - Multiple surface properties of worn RGP lenses and adhesion of Pseudomonas aeruginosa. AB - The aim of this study is to determine rigid gas permeable (RGP) lens surface properties prior to and after wear that are influential on adhesion of Pseudomonas aeruginosa. After 10 and 50 days of wear and after end-stage use, lenses were collected for determination of physico-chemical surface properties and bacterial adhesion in a parallel plate flow chamber. Water contact angles on unused RGP lenses amounted 47+/-13 degrees and were affected by wear. In addition, %O at the lens surfaces, as determined by X-ray photoelectron spectroscopy increased after use for 10 and 50 days, but decreased after end stage wear. The %N hardly increased after wear and, in line, SDS-PAGE did not indicate adsorbed proteins. The surface roughness of the lenses, as measured by atomic force microscopy amounted 9 nm after 10 and 50 days of use, but end-stage lenses were significantly rougher (48+/-23 nm). Moreover, initial deposition of P. aeruginosa #3 increased with increasing roughness for end-stage lenses. Multiple regression analysis, however, revealed that both physical and chemical surface properties were predictive for initial bacterial deposition to lens surfaces. After 10 days of wear, bacterial deposition was governed by the water contact angle, surface roughness, %O, %N, and %Si, while after 50 days of wear the surface roughness, %N, and %Si were found predictive for bacterial deposition. Initial bacterial deposition to end-stage lenses was solely dependent on the surface roughness. Summarizing, physico-chemical surface properties of RGP lenses change slightly during the first 10-50 days of wear, but end-stage lenses all had increased surface roughness, concurrent with increased bacterial adhesion. PMID- 12559827 TI - Comparison of the mechanical behaviors of biological tissues subjected to uniaxial tensile testing: pig, calf and ostrich pericardium sutured with Gore Tex. AB - The purpose of this study was to compare the mechanical behavior of calf pericardium, pig pericardium and ostrich pericardium when subjected to tensile testing. Tensile stress was applied to 108 tissue samples, 36 of each type of tissue, until rupture. Groups of three adjacent strips measuring 12 x 2 cm(2) were cut longitudinally. Each group consisted of an unsutured center sample, or control, and the two contiguous samples, that on the right sutured with Gore-Tex at a 90 degrees angle with respect to the longitudinal axis and that on the left sewn with the same suture material at 45 degrees angle. The sutured samples showed a statistically significant loss of resistance (p<0.001) when compared with the corresponding unsutured tissue. The mean stresses at rupture for sutured ostrich pericardium were 21.81 and 20.81 MPa in the samples sewn at 45 degrees and 90 degrees, respectively, higher than those corresponding to unsutured calf and pig pericardium, 14.0 and 11.49 MPa, respectively, at rupture. The analysis of the stress/strain curve shows a smaller difference between sutured and unsutured ostrich pericardium than those observed in the other two biomaterials. These results demonstrate that, in addition to its greater resistance, ostrich pericardium also presents a less pronounced interaction with the suture material. Its capacity to absorb the shearing stress produced by the suture is greater. This report also confirms that the method of selection using paired samples ensures their homogeneity and makes it possible to predict the behavior of a sample by determining that of the other half of the pair. PMID- 12559829 TI - Synaptic tagging during synapse-specific long-term facilitation of Aplysia sensory-motor neurons. AB - Like memory, long-lasting forms of synaptic plasticity have been shown to require mRNA and protein synthesis. Since each neuron has a single nucleus, but can form thousands of synaptic connections, the requirement for transcription raises the question of whether long-lasting forms of plasticity can occur in a synapse specific manner and, if so, how the products of gene expression can be targeted to alter synaptic strength at some but not all synapses made by a given neuron. One hypothesis that has been put forth to address this question is the synaptic tagging hypothesis, which proposes that the products of gene expression are delivered throughout the cell, but function to increase synaptic strength only at synapses that have been "tagged" by previous synaptic activity. In this review, I describe our studies of synapse-specific, transcription-dependent facilitation of cultured Aplysia sensory-motor synapse in the context of the synaptic tagging hypothesis. PMID- 12559830 TI - Coincident induction of long-term facilitation at sensory-motor synapses in Aplysia: presynaptic and postsynaptic factors. AB - Induction of long-term synaptic changes at one synapse can facilitate the induction of long-term plasticity at another synapse. Here we show that if Aplysia sensory neuron (SN) somata and their remote motor neuron (MN) synapses are simultaneously exposed to serotonin (5HT) pulses, which at either site alone are insufficient to induce long-term facilitation (LTF), processes activated at these sites interact to induce LTF. Coincident induction of LTF requires: (1) that the synaptic pulse occurs within a brief temporal window of the somatic pulse and (2) that local protein synthesis occurs immediately at the synapse, followed by delayed protein synthesis at the soma. LTF at the SN-MN synapses can also be induced with cell-wide application of repeated pulses of 5HT. However, these two forms of LTF differ mechanistically: (1) coincident LTF requires protein synthesis in the postsynaptic motor neuron, whereas repeated 5HT LTF does not, and (2) repeated 5HT LTF is accompanied by intermediate-term (3 h) facilitation, whereas coincident LTF is not. Thus LTF expressed in the same temporal domain can result from different underlying mechanisms. PMID- 12559831 TI - Local synthesis of proteins at synaptic sites on dendrites: role in synaptic plasticity and memory consolidation? AB - Long-lasting forms of memory are thought to be mediated by modifications in synapses that are induced by particular patterns of activity, take time to be established (consolidated), and involve new gene expression. The molecular mechanisms underlying such long-lasting synaptic modifications remain to be defined. Here, we focus on new findings regarding synapse-specific gene expression and especially the intracellular transport and synaptic targeting of the mRNA for a recently identified immediate early gene called activity-regulated cytoskeleton-associated protein (Arc). Arc mRNA is transported into dendrites after episodes of neuronal activation. The newly synthesized Arc mRNA localizes selectively at synapses that have experienced particular patterns of activity, and this targeting depends on NMDA receptor activation. Arc protein is assembled into the postsynaptic junction, and Arc expression is strongly induced by brief behavioral experiences in select populations of neurons. These features of Arc induction and trafficking reveal novel cellular mechanisms that are well suited to mediate long-term synapse-specific modifications. We will review findings from other laboratories that both LTP and memory are disrupted when Arc induction is blocked, and we will discuss the possibility that Arc protein may play a key role in activity-dependent synaptic modification. PMID- 12559832 TI - The mid-ventrolateral prefrontal cortex and active mnemonic retrieval. AB - The role of the mid-ventrolateral prefrontal cortex in memory retrieval is examined and compared with the role of the mid-dorsolateral prefrontal cortex in the monitoring of information in memory. It has been argued that the mid ventrolateral prefrontal cortex (areas 47/12 and 45) is involved in the active retrieval of information from posterior cortical association areas. Active retrieval is required when stimuli in memory do not bear stable relations to each other and therefore retrieval cannot be automatically driven by strong, stable, and unambiguous stimulus or context relations. Data from functional activation studies with normal human subjects are presented that have demonstrated specific changes in activity within the mid-ventrolateral region of the frontal cortex in relation to the active retrieval of information from memory. By contrast, increases in activity in the mid-dorsolateral region of the frontal cortex occur when the performance of the tasks requires monitoring of information in memory. PMID- 12559834 TI - Imaging brain plasticity during motor skill learning. AB - The search for the neural substrates mediating the incremental acquisition of skilled motor behaviors has been the focus of a large body of animal and human studies in the past decade. Much less is known, however, with regard to the dynamic neural changes that occur in the motor system during the different phases of learning. In this paper, we review recent findings, mainly from our own work using fMRI, which suggest that: (i) the learning of sequential finger movements produces a slowly evolving reorganization within primary motor cortex (M1) over the course of weeks and (ii) this change in M1 follows more dynamic, rapid changes in the cerebellum, striatum, and other motor-related cortical areas over the course of days. We also briefly review neurophysiological and psychophysical evidence for the consolidation of motor skills, and we propose a working hypothesis of its underlying neural substrate in motor sequence learning. PMID- 12559833 TI - Amygdala modulation of memory consolidation: interaction with other brain systems. AB - There is a strong consensus that the amygdala is involved in mediating influences of emotional arousal and stress on learning and memory. There is extensive evidence that the basolateral amygdala (BLA) is a critical locus of integration of neuromodulatory influences regulating the consolidation of several forms of memory. Many drug and stress hormone influences converge in activating the release of norepinephrine (NE) within the BLA. Evidence from studies using in vivo microdialysis and high-performance liquid chromatography indicates that increases in amygdala NE levels assessed following inhibitory avoidance training correlate highly with subsequent retention. Other evidence indicates that NE influences on memory consolidation require muscarinic cholinergic activation within the BLA provided by projections from the nucleus basalis magnocellularis (NB). Evidence from several experiments indicates that activation of the BLA plays an essential role in modulating memory consolidation processes involving other brain regions. These findings provide strong support for the hypothesis that the BLA plays a critical role in regulating the consolidation of lasting memories of significant experiences. PMID- 12559835 TI - Hippocampal memory formation, plasticity, and the role of sleep. PMID- 12559836 TI - Gene x environment interaction and cognitive performance: animal studies on the role of corticosterone. AB - A fundamental question in the neurobiology of cognition is how stress and glucocorticoids modify learning and memory processes. Why some individuals develop cognitive deficits after stress, while other individuals improve in cognitive performance under similar adverse conditions is still unresolved. To address these questions we focus on those issues. First, corticosterone, which appears to be the preferred glucocorticoid for the rodent and human brain, acts via brain mineralocorticoid (MR) and glucocorticoid receptors (GR) on the expression of networks of corticosteroid-responsive genes. Different effects are achieved by MR and GR activation. Second, the experimental context that determines the timing and the consequences of corticosterone action during the various stages of information processing is reviewed. Third, the genetic context and the environmental context are investigated. Using apolipoprotein E knockout (apoE0/0) mice we show that apoE (apoE4 is a genetic risk factor for Alzheimer's disease) is a candidate gene with an important function in shaping the cognitive outcome (genotype x environment interaction). PMID- 12559837 TI - Stress and memory: opposing effects of glucocorticoids on memory consolidation and memory retrieval. AB - It is well established that glucocorticoid hormones, secreted by the adrenal cortex after a stressful event, influence cognitive performance. Some studies have found glucocorticoid-induced memory enhancement. However, many studies have reported impairing effects of glucocorticoids on memory function. This paper reviews recent findings from this laboratory on the acute effects of glucocorticoids in rats on specific memory phases, i.e., memory consolidation and memory retrieval. The evidence suggests that the consequences of glucocorticoid activation on cognition depend largely on the different memory phases investigated. Posttraining activation of glucocorticoid-sensitive pathways involving glucocorticoid receptors enhances memory consolidation in a pattern highly similar to that previously described for adrenal catecholamines. Also, similar to catecholamine effects on memory consolidation, glucocorticoid influences on memory consolidation depend on noradrenergic activation of the basolateral complex of the amygdala and interactions with other brain regions. By contrast, memory retrieval processes are usually impaired with high circulating levels of glucocorticoids or following infusions of glucocorticoid receptor agonists into the hippocampus. The hypothesis is proposed that these apparently dual effects of glucocorticoids on memory consolidation and memory retrieval might be related and that the basolateral complex of the amygdala is a key structure in a memory-modulatory system that regulates, in concert with other brain regions, stress and glucocorticoid effects on both memory consolidation and memory retrieval. PMID- 12559838 TI - Effects of stress hormones on traumatic memory formation and the development of posttraumatic stress disorder in critically ill patients. AB - A majority of patients after intensive care treatment report traumatic memories from their stay in the intensive care unit (ICU). Traumatic memories can be associated with the development of posttraumatic stress disorder (PTSD) in a subpopulation of these patients. In contrast to other patient populations at risk for PTSD, patients in the ICU often receive exogenously administered stress hormones like epinephrine, norepinephrine, or cortisol for medical reasons and are extensively monitored. ICU patients therefore represent a suitable population for studying the relationship between stress hormones, traumatic memories, and the development of PTSD. Studies in long-term survivors of ICU treatment demonstrated a clear and vivid recall of different categories of traumatic memory such as nightmares, anxiety, respiratory distress, or pain with little or no recall of factual events. The number of categories of traumatic memory recalled increased with the total administered dosages of stress hormones (both catecholamines and cortisol), and the evaluation of these categories at different time points after discharge from the ICU showed better memory consolidation with higher dosages of stress hormones administered. However, the administration of stress doses of cortisol to critically ill patients resulted in more complex findings as it caused a significant reduction in PTSD symptoms measured after recovery. This effect can possibly be explained by a differential influence of cortisol on memory. Increased serum cortisol levels not only result in consolidation of emotional memory but are also known to cause a temporary impairment in memory retrieval which appears to be independent of glucocorticoid effects on memory formation. Disrupting retrieval mechanisms with glucocorticoids during critical illness may therefore act protectively against the development of PTSD by preventing recall of traumatic memories. Our findings indicate that stress hormones influence the development of PTSD through complex and simultaneous interactions on memory formation and retrieval. Our studies also demonstrate that animal models of aversive learning are useful in analyzing and predicting clinical findings in critically ill humans. PMID- 12559839 TI - Role of dopamine, the frontal cortex and memory circuits in drug addiction: insight from imaging studies. AB - Drug addiction is characterized by a set of recurring processes (intoxication, withdrawal, craving) that lead to the relapsing nature of the disorder. We have used positron emission tomography to investigate in humans the role of dopamine (DA) and the brain circuits it regulates in these processes. We have shown that increases in DA are associated with the subjective reports of drug reinforcement corroborating the relevance of drug-induced DA increases in the rewarding effects of drugs in humans. During withdrawal we have shown in drug abusers significant reductions in DA D2 receptors and in DA release. We postulate that this hypodopaminergic state would result in a decreased sensitivity to natural reinforcers perpetuating the use of the drug as a means to compensate for this deficit and contributing to the anhedonia and dysphoria seen during withdrawal. Because the D2 reductions are associated with decreased activity in the anterior cingulate gyrus and in the orbitofrontal cortex we postulate that this is one of the mechanisms by which DA disruption leads to compulsive drug administration and the lack of control over drug intake in the drug-addicted individual. This is supported by studies showing that during craving these frontal regions become hyperactive in proportion to the intensity of the craving. Craving is also associated with activation of memory circuits including the amygdala (implicated in conditioned learning), hippocampus (implicated in declarative learning), and dorsal striatum (implicated in habit learning) all of which receive DA innervation. We therefore postulate that dopamine contributes to addiction by disrupting the frontal cortical circuits that regulate motivation, drive, and self-control and by memory circuits that increase the motivational salience of the drug and drug-associated stimuli. PMID- 12559840 TI - Limbic-striatal memory systems and drug addiction. AB - Drug addiction can be understood as a pathological subversion of normal brain learning and memory processes strengthened by the motivational impact of drug associated stimuli, leading to the establishment of compulsive drug-seeking habits. Such habits evolve through a cascade of complex associative processes with Pavlovian and instrumental components that may depend on the integration and coordination of output from several somewhat independent neural systems of learning and memory, each contributing to behavioral performance. Data are reviewed that help to define the influences of conditioned Pavlovian stimuli on goal-directed behavior via sign-tracking, motivational arousal, and conditioned reinforcement. Such influences are mediated via defined corticolimbic-striatal systems converging on the ventral striatum and driving habit-based learning that may depend on the dorsal striatum. These systems include separate and overlapping influences from the amygdala, hippocampus, and cingulate and medial prefrontal cortex on drug-seeking as well as drug-taking behavior, including the propensity to relapse. PMID- 12559841 TI - Common molecular and cellular substrates of addiction and memory. AB - Drugs of abuse cause long-lasting changes in the brain that underlie the behavioral abnormalities associated with drug addiction. Similarly, experience can induce memory formation by causing stable changes in the brain. Over the past decade, the molecular and cellular pathways of drug addiction, on the one hand, and of learning and memory, on the other, have converged. Learning and memory and drug addiction are modulated by the same neurotrophic factors, share certain intracellular signaling cascades, and depend on activation of the transcription factor CREB. They are associated with similar adaptations in neuronal morphology, and both are accompanied by alterations in synaptic plasticity (e.g., long-term potentiation, long-term depression) at particular glutamatergic synapses in the brain. There has also been recent convergence in the brain regions now considered important sites for molecular and cellular plasticity underlying addiction and memory. Complex circuits involving the hippocampus, cerebral cortex, ventral and dorsal striatum, and amygdala are implicated both in addiction and in learning and memory. The complexity of the plasticity that occurs in these circuits can be illustrated by CREB, which induces very different behavioral effects in these various brain regions. A better understanding of the molecular and cellular adaptations that occur in these neural circuits may lead to novel interventions to improve memory and combat addiction in humans. PMID- 12559842 TI - Mice deficient for cytosolic thymidine kinase gene develop fatal kidney disease. AB - The thymidine kinase (Tk) gene codes for a cytosolic protein involved in the pyrimidine nucleotide salvage pathway. A functional Tk gene is not necessary for cells in culture, and a naturally occurring Tk deficient phenotype has not been described in humans or animal models. In order to determine the biological significance of the Tk gene, we created Tk(-/-) knockout (KO) mice through homologous recombination in mouse embryonic stem cells. Tk KO mice have shortened life spans compared with their wild-type or Tk heterozygous (HET) siblings. All Tk KO mice develop sclerosis of kidney glomeruli and die before one year of age of kidney failure. Among other changes in KO animals, the most consistent is a switch from exclusively mucous secretion to predominantly serous secretion in the sublingual salivary gland. HET parents can produce KO mice at a frequency approaching Mendelian inheritance. Other observations in KO animals include an elevated level of serum thymidine, a significant decrease in the cloning efficiency of splenic lymphocytes, an increase in the frequency of hypoxanthine guanine phosphoribosyl transferase gene mutant lymphocytes, and histological alteration in the lymphoid structure of the spleen. In addition, KO animals sporadically exhibit inflammation of the arteries, which taken together with the lymphocyte and spleen abnormalities, suggest an abnormal immune system. Alterations in Tk KO mice indicate that the pyrimidine nucleotide salvage pathway is indispensable in vivo. PMID- 12559843 TI - A mouse model of argininosuccinic aciduria: biochemical characterization. AB - Argininosuccinate lyase (AL) has several roles in intermediary metabolism. It is an essential component of the urea cycle, providing a pathway for the disposal of excess nitrogen in mammals. AL links the urea cycle to the tricarboxylic acid (TCA) cycle by generating fumarate. Finally, AL is required for the endogenous production of arginine. In this latter role it may function outside ureagenic organs to provide arginine as a substrate for nitric oxide synthases (NOS). Increasing evidence suggests that argininosuccinate synthetase (AS) and AL are more globally expressed, and the coordinate regulation of AS and AL gene expression with that of the inducible form of NOS (iNOS) provides evidence that this may facilitate the regulation of NOS activity. Deficiency of AL leads to the human urea cycle disorder argininosuccinic aciduria. We produced an AL deficient mouse by gene targeting in order to investigate the role of AL in endogenous arginine production. This mouse also provides a model of the human disorder to explore the pathogenesis of the disorder and possible new treatments. Metabolic studies of these mice demonstrated that they have the same biochemical phenotype as humans, with hyperammonemia, elevated plasma argininosuccinic acid and low plasma arginine. Plasma nitrites, derived from NO, were not reduced in AL deficient mice and there was no significant difference is the level of cyclic GMP, the second messenger induced by NO. PMID- 12559844 TI - Functional categorization of gene expression changes in the cerebellum of a Cln3 knockout mouse model for Batten disease. AB - Juvenile neuronal ceroid lipofuscinosis (JNCL or Batten Disease) is the most common progressive neurodegenerative disorder of childhood. The disease is inherited in an autosomal recessive manner and is the result of mutations in the CLN3 gene. One brain region severely affected in Batten disease is the cerebellum. Using a mouse model for Batten disease which shares pathological similarities to the disease in humans we have used oligonucleotide arrays to profile approximately 19000 mRNAs in the cerebellum. We have identified reproducible changes of twofold or more in the expression of 756 gene products in the cerebellum of 10-week-old Cln3-knockout mice as compared to wild-type controls. We have subsequently divided these genes with altered expression into 14 functional categories. We report a significant alteration in expression of genes associated with neurotransmission, neuronal cell structure and development, immune response and inflammation, and lipid metabolism. An apparent shift in metabolism toward gluconeogenesis is also evident in Cln3-knockout mice. Further experimentation will be necessary to understand the contribution of these changes in expression to a disease state. Detailed analysis of the functional consequences of altered expression of genes in the cerebellum of the Cln3 knockout mice may provide valuable clues in understanding the molecular basis of the pathological mechanisms underlying Batten disease. PMID- 12559845 TI - Visceral obesity attenuates the effect of the hepatic lipase -514C>T polymorphism on plasma HDL-cholesterol levels in French-Canadian men. AB - The dyslipidemic state of visceral obesity is characterized by increased plasma triglyceride (TG) levels, low HDL-cholesterol concentrations and alterations in LDL composition and concentration. A functional, non-coding -514C>T single nucleotide polymorphism (SNP) of the hepatic lipase gene (LIPC) has been related to variation in HDL-cholesterol concentrations. OBJECTIVES: To investigate the hypotheses that the LIPC -514C>T polymorphism may be associated with a deteriorated lipoprotein-lipid profile and that environmental factor, such as abdominal obesity, alters this association. METHODS: A total of 235 French Canadian men from the greater Quebec City area were assigned into three groups on the basis of their LIPC -514C>T SNP, including 149 CC homozygotes, 75 CT heterozygotes, and 11 TT homozygotes. RESULTS: In the present study, the highest values of BMI, waist circumference, and accumulation of visceral adipose tissue (VAT) were observed among TT homozygotes (p<0.05). After adjustment for age and BMI, TT homozygotes still showed higher plasma apolipoprotein (apo) AI and HDL-TG concentrations than the two other groups (p<0.05). When the two genotype groups (CC vs CT/TT) were further divided on the basis of VAT accumulation using a cut off point of 130 cm(2) (high vs low) it appears that irrespective of the genotype subjects with low VAT had higher HDL(2)-cholesterol concentrations (p<0.0001). However, lean carriers of the T allele had higher plasma HDL(2)-cholesterol levels than lean CC homozygotes. The beneficial effect of the T allele on plasma HDL(2)-cholesterol levels was abolished in the presence of visceral obesity (VAT>130 cm(2)). CONCLUSION: In summary, the presence of visceral obesity attenuates the impact of the LIPC -514C>T polymorphism on plasma HDL(2) cholesterol levels. PMID- 12559846 TI - Identification and characterization of 13 new mutations in mucopolysaccharidosis type I patients. AB - In this study we have investigated a group of 29 Brazilian patients, who had been diagnosed with the lysosomal storage disorder, Mucopolysaccharidosis type I (MPS I). MPS I is caused by a deficiency in the lysosomal hydrolase, alpha-L iduronidase. Ninety percent of the MPS I patients in this study were genotyped and revealed 10 recurrent and thirteen novel IDUA gene mutations. Eight of these new mutations and three common mutations W402X, P533R, and R383H were individually expressed in CHO-K1 cells and analyzed for alpha-L-iduronidase protein and enzyme activity. A correlation was observed between the MPS I patient clinical phenotype and the associated mutant alpha-L-iduronidase protein/enzyme activity expressed in CHO-K1 cells. This was the first time that Brazilian MPS I patients had been thoroughly analyzed and highlighted the difficulties of mutation screening and clinical phenotype assessment in populations with high numbers of unique mutations. PMID- 12559847 TI - The AGT gene in Africa: a distinctive minor allele haplotype, a polymorphism (V326I), and a novel PH1 mutation (A112D) in Black Africans. AB - We describe a novel missense mutation (A112D) and polymorphism (V326I) in the human AGT gene in two black African patients with primary hyperoxaluria type 1, an autosomal recessive disease resulting from a deficiency of the liver peroxisomal enzyme alanine:glyoxylate aminotransferase (AGT; EC 2.6.1.44). V326I was found in DNA from normal control Blacks with an allele frequency of 3%. Expression studies confirmed that A112D reduced AGT enzyme activity by 95% while V326I had no effect. Both A112D and V326I were homozygous in both patients and lie on a variant of the minor allele of the AGT gene. This variant haplotype, Mi(A), includes an intron 1 duplication and intron 4 VNTR (38 repeat) but lacks the P11L and I340M normally associated with the minor allele in Caucasians. Among the South African Blacks tested, the Mi(A) haplotype had an allele frequency of 12% compared to 3 % for the Caucasian-type minor allele haplotype. PMID- 12559848 TI - Imbalanced substrate specificity of mutant beta-galactosidase in patients with Morquio B disease. AB - G(M1)-gangliosidosis and Morquio B disease are distinct in clinical and biochemical features, but both disorders are caused by genetic defects of the same enzyme, acid beta-galactosidase (beta-Gal). We analyzed the kinetic properties of mutant beta-Gals from patients with G(M1)-gangliosidosis and Morquio B disease to examine the clinical and biochemical differences between both disorders. Five skin fibroblast lines from patients with G(M1) gangliosidosis (2 cases; R201C/R201C and I51T/I51T), Morquio B disease (2 cases; W273L/W273L and Y83H/R482C), and galactosialidosis (1 case; Y395C/S90L) were used as enzyme sources. Residual enzyme activity in the cells was subjected to kinetic analysis. Substrate analogs including Galbeta1-3GalNAc, as an analog for G(M1) ganglioside, and Galbeta1-4GlcNAc, as an analog for keratan sulfate, were used to determine IC(50) and K(i) for beta-Gals with an artificial substrate (4 methylumbelliferyl beta-D-galactopyranoside). Enzymatic assay method was established to examine the hydrolytic activity with the mutant beta-Gal for the substrate analogs. The mutant beta-Gal activities were inhibited by Galbeta1 3GalNAc and Galbeta1-4GlcNAc in a concentration-dependent manner. Remarkable increase in IC(50) ratio and K(i) ratio (Galbeta1-4GlcNAc/Galbeta1-3GalNAc) was observed in Morquio B disease. Relative hydrolytic activity (Galbeta1 4GlcNAc/Galbeta1-3GalNAc) was markedly decreased in Morquio B disease as compared with other subjects; controls (means+/-SD, n=4), 1.00+/-0.02; galactosialidosis, 1.03; G(M1)-gangliosidosis, 1.15 and 1.00; and Morquio B disease, 0.27 and 0.32. The mutant beta-Gals from the patients with Morquio B disease exhibited lower affinity and lower hydrolytic activity toward Galbeta1-4GlcNAc rather than Galbeta1-3GalNAc. These findings suggest that imbalanced substrate specificity of the mutant beta-Gals induces predominant accumulation of keratan sulfate and a rationale for performing differential diagnostic analysis for both disorders. PMID- 12559849 TI - Propionic acidemia: identification of twenty-four novel mutations in Europe and North America. AB - Propionic acidemia is an inherited metabolic disease caused by the deficiency of the mitochondrial protein propionyl-CoA carboxylase (PCC), one of the four biotin dependent enzymes. PCC is a multimeric protein composed of two different alpha- and beta-PCC subunits, nuclearly encoded by the PCCA and PCCB genes, respectively. Mutations in either gene cause the clinically heterogeneous disease propionic acidemia. In this work we describe the mutational analysis of PCCA and PCCB deficient patients from different European countries (Spain, Italy, Belgium, Croatia, and Austria) and from America (mainly USA). We report 24 novel PA mutations, nine affecting the PCCA gene and 15 affecting the PCCB gene. They include six missense mutations, one nonsense mutation, one point exonic mutation affecting splicing, seven splicing mutations affecting splice sequences, and nine short insertions or deletions, only two in-frame. We have found a highly heterogenous spectrum of PCCA mutations, most of the PCCA deficient patients are homozygous carrying a unique genotype. The PCCA mutational spectrum includes a high proportion of short insertions or deletions affecting one nucleotide. In the PCCA mutant alleles analyzed we have also found one single nucleotide change, a novel nonsynonymous SNP. On the other hand, the PCCB deficient patients carry a more reduced spectrum of mutations, 50% of them are missense. This work represents an extensive update of the mutational study of propionic acidemia providing important information about the worldwide distribution of PA mutations and representing another essential part in the study of the phenotype-genotype correlations for the prediction of the metabolic outcome and for the implementation of treatments tailored to each PA patient. PMID- 12559850 TI - Mutational spectrum and DNA-based prenatal diagnosis in carnitine-acylcarnitine translocase deficiency. AB - Carnitine-acylcarnitine translocase (CAC) deficiency is a rare autosomal recessive disorder of long-chain fatty acid oxidation with a severe outcome. We report mutation analysis in a cohort of 12 patients. Twelve mutations were identified of which 9 have not been reported so far (G28C, D32N, R178Q, P230R, D231H, 179delG, 802delG, 69-70insTGTGC, and 609-1g>a). Altogether, including our results, 22 mutations of the CAC gene have been published to date in 23 patients demonstrating the allelic heterogeneity of CAC deficiency. DNA-based prenatal diagnosis was performed for the first time in pregnancies at risk for CAC deficiency. Two fetuses were affected and one pregnancy was terminated by family decision. Two other fetuses had normal genotype and five others were heterozygotes. All the offspring of these seven pregnancies are alive and apparently healthy. PMID- 12559851 TI - Lithium detaches hexokinase from mitochondria and inhibits proliferation of B16 melanoma cells. AB - Glycolysis is known to be the primary energy source in cancer cells. Hexokinase (ATP: D-hexose 6-phosphotransferase, EC 2.7.1.1), the only glycolytic enzyme which binds to mitochondria, is exceptionally high in cancer cells, and believed to play a key role in regulating cell energy metabolism and cancer cell growth rate. We show here that lithium induces a detachment of hexokinase from mitochondria of B16 melanoma cells. This effect eventually leads to inhibition of cell proliferation. These results reveal a novel, additional, mechanism of action of lithium and suggest that lithium may be promising drug in treatment of melanoma. PMID- 12559852 TI - A novel single base deletion at codon 434 (1301delT) of the DAX1 gene associated with prepubertal testis enlargement. AB - We have identified a novel DAX1 frameshift mutation (1301delT) at codon 434 in a patient with primary adrenal insufficiency. This 11-day-old boy was admitted to the hospital with hyponatremia, hyperkalemia, and suspected congenital adrenal abnormality. He exhibited severe hypoglycemia, pallor of the skin, buccal and genital hyperpigmentation, hypotension (90/45 mm Hg), anemia, and diarrhea. Although basal gonadotropins were low, and responded minimally to exogenous GnRH, the size of his testes began to increase at age 4 and reached 4.5 mL at the age of 9 years and 8 months. Testosterone levels were prepubertal. These findings further emphasize the variable clinical presentation in children with DAX1 gene mutations and indicate the value of genetic testing in boys with primary adrenal insufficiency. PMID- 12559854 TI - Whose interests does the World Trade Organization serve? PMID- 12559855 TI - A neuropathology of psychosis? PMID- 12559856 TI - What puts children of lone parents at a health disadvantage? PMID- 12559857 TI - Dominant SUR1 mutation causing autosomal dominant type 2 diabetes. PMID- 12559858 TI - In-vitro fertilisation and retinoblastoma. PMID- 12559859 TI - Another loss in the privatisation war: PubScience. PMID- 12559860 TI - Recombinant human interleukin 11 and bacterial infection in patients with [correction of] haematological malignant disease undergoing chemotherapy: a double-blind placebo-controlled randomised trial. AB - BACKGROUND: Bacteraemia in patients with haematological malignant disease causes substantial morbidity. Recombinant human interleukin 11 (rhIL-11) prevents gastrointestinal epithelial disintegrity and has immunomodulatory actions. Our aim was to ascertain whether or not treatment with rhIL-11 can prevent gut associated infections. METHODS: We did a double-blind placebo-controlled randomised trial, to which we enrolled 40 patients with haematological malignant disease who were undergoing chemotherapy. Patients received either rhIL-11 50 microg/kg (n=20) or placebo (n=20) daily by subcutaneous injection from the day before the start of chemotherapy until resolution of neutropenia or for 21 days, whichever was longer. Our primary outcome measure was a reduction in bacteraemia. Analysis was by intention to treat. FINDINGS: Significantly fewer patients who received rhIL-11 rather than placebo developed bacteraemia, particularly of gastrointestinal origin: the proportion of patients with at least one positive blood culture was 0.65 and 0.25, respectively (p=0.02). The numbers of patients (placebo vs rhIL-11) for each number of distinct isolates were: no organism isolated seven versus 15, one organism nine versus four, two organisms two versus one, three organisms one versus none, and four organisms one versus none (p=0.01), suggesting a lower bacterial load in the rhIL-11 than in the placebo group. Time to first bacteraemic event was longer in patients who received rhIL 11 (p=0.03) than in those who received placebo. INTERPRETATION: rhIL-11 reduces the frequency and load of bacteraemia in patients with haematological malignant disease undergoing chemotherapy, possibly by gastrointestinal cytoprotective or immunological mechanisms [corrected]. PMID- 12559861 TI - Neuroanatomical abnormalities before and after onset of psychosis: a cross sectional and longitudinal MRI comparison. AB - BACKGROUND: Psychotic disorders, such as schizophrenia, are associated with neuroanatomical abnormalities, but whether these predate the onset of symptoms or develop progressively over the course of illness is unclear. We investigated this issue with MRI to study people with prodromal symptoms who are at ultra high-risk for the development of psychosis. METHODS: We did two comparisons, cross sectional and longitudinal. For the cross-sectional comparison, 75 people with prodromal signs of psychosis were scanned with MRI. After at least 12 months of follow-up, 23 (31%) had developed psychosis and 52 (69%) had not. Baseline MRI data from these two subgroups were compared. For the longitudinal comparison, 21 of the ultra high-risk individuals were scanned again with MRI after at least 12 months. Ten of these had developed psychosis and 11 had not. MRI data from baseline and follow-up were compared within each group of people. FINDINGS: In the cross-sectional comparison, compared with people who did not develop psychosis, those who did develop the disorder had less grey matter in the right medial temporal, lateral temporal, and inferior frontal cortex, and in the cingulate cortex bilaterally. In the longitudinal comparison, when re-scanned, individuals who had developed psychosis showed a reduction in grey matter in the left parahippocampal, fusiform, orbitofrontal and cerebellar cortices, and the cingulate gyri. In those who had not become psychotic, longitudinal changes were restricted to the cerebellum. INTERPRETATION: Some of the grey-matter abnormalities associated with psychotic disorders predate the onset of frank symptoms, whereas others appear in association with their first expression. PMID- 12559862 TI - Mortality, severe morbidity, and injury in children living with single parents in Sweden: a population-based study. AB - BACKGROUND: Growing up with one parent has become increasingly common, and seems to entail disadvantages in terms of socioeconomic circumstances and health. We aimed to investigate differences in mortality, severe morbidity, and injury between children living in households with one adult and those living in households with two adults. METHODS: In this population-based study, we assessed overall and cause-specific mortality between 1991 and 1998 and risk of admission between 1991 and 1999 for 65085 children with single parents and 921257 children with two parents. We estimated relative risks by Poisson regression, adjusted for factors that might be presumed to select people into single parenthood, and for other factors, mainly resulting from single parenthood, that might have affected the relation between type of parenting and risk. FINDINGS: Children with single parents showed increased risks of psychiatric disease, suicide or suicide attempt, injury, and addiction. After adjustment for confounding factors, such as socioeconomic status and parents' addiction or mental disease, children in single parent households had increased risks compared with those in two-parent households for psychiatric disease in childhood (relative risk for girls 2.1 [95% CI 1.9-2.3] and boys 2.5 [2.3-2.8]), suicide attempt (girls 2.0 [1.9-2.2], boys 2.3 [2.1-2.6]), alcohol-related disease (girls 2.4 [2.2-2.7], boys 2.2 [2.0 2.4]), and narcotics-related disease (girls 3.2 [2.7-3.7], boys 4.0 [3.5-4.5]). Boys in single-parent families were more likely to develop psychiatric disease and narcotics-related disease than were girls, and they also had a raised risk of all-cause mortality. CONCLUSIONS: Growing up in a single-parent family has disadvantages to the health of the child. Lack of household resources plays a major part in increased risks. However, even when a wide range of demographic and socioeconomic circumstances are included in multivariate models, children of single parents still have increased risks of mortality, severe morbidity, and injury. PMID- 12559864 TI - A mediastinal tumour. PMID- 12559863 TI - Overview of the main outcomes in breast-cancer prevention trials. AB - BACKGROUND: Early findings on the use of tamoxifen or raloxifene as prophylaxis against breast cancer have been mixed; we update available data and overview the combined results. METHODS: All five randomised prevention trials comparing tamoxifen or raloxifene with placebo were included. Relevant data on contralateral breast tumours and side-effects were included from an overview of adjuvant trials of tamoxifen versus control. FINDINGS: The tamoxifen prevention trials showed a 38% (95% CI 28-46; p<0.0001) reduction in breast-cancer incidence. There was no effect for breast cancers negative for oestrogen receptor (ER; hazard ratio 1.22 [0.89-1.67]; p=0.21), but ER-positive cancers were decreased by 48% (36-58; p<0.0001) in the tamoxifen prevention trials. Age had no apparent effect. Rates of endometrial cancer were increased in all tamoxifen prevention trials (consensus relative risk 2.4 [1.5-4.0]; p=0.0005) and the adjuvant trials (relative risk 3.4 [1.8-6.4]; p=0.0002); no increase has been seen so far with raloxifene. Venous thromboembolic events were increased in all tamoxifen studies (relative risk 1.9 [1.4-2.6] in the prevention trials; p<0.0001) and with raloxifene. Overall, there was no effect on non-breast-cancer mortality; the only cause showing a mortality increase was pulmonary embolism (six vs two). INTERPRETATION: The evidence now clearly shows that tamoxifen can reduce the risk of ER-positive breast cancer. New approaches are needed to prevent ER-negative breast cancer and to reduce the side-effects of tamoxifen. Newer agents such as raloxifene and the aromatase inhibitors need to be evaluated. Although tamoxifen cannot yet be recommended as a preventive agent (except possibly in women at very high risk with a low risk of side-effects), continued follow-up of the current trials is essential for identification of a subgroup of high-risk, healthy women for whom the risk-benefit ratio is sufficiently positive. PMID- 12559865 TI - A new subtype of autosomal dominant diabetes attributable to a mutation in the gene for sulfonylurea receptor 1. AB - BACKGROUND: ATP-sensitive potassium (KATP) channels are major regulators of glucose-induced insulin secretion in pancreatic beta cells. We have described a dominant heterozygous mutation--E1506K--in the sulfonylurea receptor 1 (SUR1) gene (ABCC8) in a Finnish family, which leads to congenital hyperinsulinaemia due to reduction of K(ATP)-channel activity. We aimed to characterise glucose metabolism in adults heterozygous for the E1506K mutation. METHODS: Glucose tolerance was assessed by an oral glucose tolerance test, insulin secretion by the intravenous glucose tolerance test and hyperglycaemic clamp, and insulin sensitivity by hyperinsulinaemic euglycaemic clamp in 11 people heterozygous for the E1506K mutation and 19 controls. FINDINGS: Four people who were heterozygous for the SUR1 E1506K mutation had diabetes, five had impaired glucose tolerance, one had impaired fasting glucose, and one had normal glucose tolerance. Although glucose-induced, first-phase insulin secretion was normal in children younger than 10 years of age who were heterozygous for the SUR1 E1506K mutation (n=2; 66 and 334 pmol/L), it fell rapidly after puberty (n=3; 12-32 pmol/L), and was almost completely lost in adulthood (n=11; 12-32 pmol/L). Furthermore, these heterozygous people had a substantial reduction in maximum glucose-stimulated insulin secretion during hyperglycemic clamp (carriers without diabetes 422 pmol/L; carriers with diabetes 97 pmol/L). By contrast, insulin sensitivity (M/I value) was normal in carriers of the E1506K mutation who did not have diabetes and was reduced by 15% in those who were heterozygous with diabetes (0.07 in those without diabetes and 0.05 in those with the disorder; not significantly different from controls). INTERPRETATION: Heterozygous E1506K substitution in the SUR1 gene causes congenital hyperinsulinism in infancy, loss of insulin secretory capacity in early adulthood, and diabetes in middle-age. This variant represents a new subtype of autosomal dominant diabetes. PMID- 12559866 TI - Agitation by sedation. PMID- 12559867 TI - Incidence of retinoblastoma in children born after in-vitro fertilisation. AB - Every year, 3000 women in the Netherlands are treated with in-vitro fertilisation (IVF), and results from studies suggest that the offspring of these women do not have a heightened risk of cancer. Between November, 2000, and February, 2002, we diagnosed retinoblastoma in five patients born after IVF. To calculate the relative risk of the disease, we assumed that the proportion of children conceived by IVF in the Netherlands is between 1.0 and 1.5%, and that the five patients who we diagnosed with the disease represent all new cases in the Netherlands during that period. Relative risks for retinoblastoma were significantly raised (7.2 [95% CI 2.4-17.0], and 4.9 [1.6-11.3], for 1% and 1.5% rates, respectively). This possible association of an increased risk of retinoblastoma in a population-based study needs to be established. PMID- 12559868 TI - Faecal calprotectin concentrations and diagnosis of necrotising enterocolitis. AB - Calprotectin has been proposed as a useful marker of inflammatory bowel disease in children. We did a pilot study to establish whether it can be used to aid diagnosis of necrotising enterocolitis in preterm infants. Patients with clinical features of necrotising enterocolitis had raised faecal calprotectin concentrations at the time of diagnosis compared with matched controls (288.4 mg/L [SD 49.1] and 98.0 mg/L [60.6], respectively; p=0.0006). Faecal calprotectin might be a useful marker of gastrointestinal mucosal inflammation in neonates. PMID- 12559869 TI - Syringes with rubber-coated plungers and inactivation of surfactant. AB - Injectable drugs are generally administered to newborns with 1 mL syringes. During in-vitro measurements of surface tension with a pulsating bubble surfactometer, we noticed that surfactant was inactivated after repeated aspiration into a 1 mL syringe with a rubber-coated plunger. Inactivation did not take place, however, when we used rubber-free two-part syringes with polyethylene pistons. Results of further studies showed that the silicone fluid used to lubricate the rubber surface of the syringe was inactivating the surfactant. Our findings suggest that contact with rubber surfaces should be avoided in the handling of surfactant material for biophysical assessment or clinical use. PMID- 12559870 TI - Targeted mass treatment for syphilis with oral azithromycin. AB - From mid 1997 to end of 1999, there was a sexually-transmitted infectious syphilis outbreak mainly in heterosexual people in British Columbia, Canada, that was concentrated in Vancouver. The rate across the province increased from less than 0.5 to 3.4 per 100000, and the rate in Vancouver reached 12.9 per 100000. We aimed to eliminate the syphillis outbreak by treating people at risk of infection. In 2000, a targeted mass treatment programme provided azithromycin (1.8 g orally) to 4384 at-risk residents in this city. After the programme, syphilis frequency fell significantly for 6 months (p=0.016), but rose again in 2001. Results from curve fitting analyses showed that the number of cases in 2001 (177) was higher than expected (0.00013 h after the onset of symptoms, the mortality of the TL group reached 15.3% compared to 6% in the PCI group (P<0.02). Patients randomized within <3 h of symptom onset (n=551) had no difference in mortality whether treated by TL (7.4%) or transferred to PCI (7.3%). A combined end-point occurred in 15.2% of the TL group vs 8.4% of the PCI group (P<0.003). CONCLUSIONS: Long distance transport from a community hospital to a tertiary PCI centre in the acute phase of AMI is safe. This strategy markedly decreases mortality in patients presenting >3 h after symptom onset. For patients presenting within <3 h of symptoms, TL results are similar results to long distance transport for PCI. PMID- 12559942 TI - The use of hospital admission data as a measure of outcome in clinical studies of heart failure. AB - AIM: To describe and evaluate the methods currently used to summarize, depict, and analyse hospitalization data in clinical studies of patients with heart failure. METHODS AND RESULTS: Studies published during 1999 and 2000 were reviewed if they reported hospitalization data collected during the follow-up of a defined group of adults who had been diagnosed as having heart failure. Of 130 studies identified, the majority considered only the first of any hospital admissions for each individual. When presented, statistical analyses in 70% of papers, and graphics in 75% of papers, were based purely on the first hospitalization. A minority of papers based analyses on the number of admissions (23%), or the amount of time spent in hospital by patients (14%) during the follow-up period. CONCLUSIONS: Data on the hospitalizations undergone by patients with heart failure are potentially informative about the disease burden to individuals and health services. The widespread practice of focussing a statistical analysis on the first admission a patient undergoes discards those aspects of hospitalization that are informative about disease burden: multiple admissions and length of stay. Statistical methods that incorporate these aspects appeared in a handful of papers, and the more widespread adoption of these is desirable. PMID- 12559943 TI - Tissue Doppler echocardiography in patients with thalassaemia detects early myocardial dysfunction related to myocardial iron overload. AB - AIMS: To compare an echocardiographic method for detecting abnormal cardiac function before development of overt cardiomyopathy with a recently validated technique of quantifying myocardial iron load. METHODS AND RESULTS: We examined thalassaemia patients whose myocardial iron load had been evaluated with magnetic resonance imaging (MRI). By tissue Doppler echocardiography, myocardial velocities were sampled continuously from base to apex in the RV and LV free wall, and the septum in 52 patients aged 29.2 (14.2-43.1) years and 52 age matched controls. Ninety-six percent of patients had normal LV ejection fraction by MRI. Thirty-eight (73%) had abnormal iron loading of the myocardium, and 33 of those had regional wall motion abnormalities detected in the septum (n=29), LV (n=2), RV (n=1), and septum plus LV (n=1). The incidence of wall motion abnormalities was significantly higher (P<0.04) in patients with myocardial iron overload (87%) than in the 14 without (35%). Furthermore, myocardial iron overload was suggested by a low T2(*)(15.1+/-15.8 ms) in patients with wall motion abnormalities vs those with normal wall motion (T2(*): 30+/-19 ms) (P<0.007). CONCLUSIONS: Wall motion abnormalities may represent an early sign of cardiac disease despite preserved global function. The regional abnormalities are related to iron overload and easily detectable with tissue Doppler echocardiography. PMID- 12559944 TI - Phosphorylation of serine 276 is essential for p65 NF-kappaB subunit-dependent cellular responses. AB - Phosphorylation of several serine residues especially in the transactivation (TA) domain of p65 NF-kappaB subunit has been suggested to be important for its transcriptional activity. However, the responsible phosphorylation site of p65 remains controversial. To investigate the biological significance of phosphorylation and to determine the critical phosphorylation sites of p65, we reconstituted murine embryonic fibroblasts (MEFs) from p65(-/-) mice with various serine to alanine (SA)-substituted mutants of p65. Unexpectedly, mutants in the TA domain, including S529A, S536A, and S529A/S536A, completely rescued the defect of p65(-/-) MEFs as assessed by tumor necrosis factor (TNF)- or interleukin-1 (IL 1)-induced IL-6 production and protection from TNF-induced cell death. On the other hand, S276A mutant had an impaired ability to rescue these responses. Moreover, TNF-induced phosphorylation of p65 was severely impaired in S276A mutant, indicating that S276 is the major phosphorylation site of p65 and its phosphorylation is essential for p65-dependent cellular responses. PMID- 12559945 TI - Homeodomain protein CDX2 regulates goblet-specific MUC2 gene expression. AB - Intestinal mucin gene MUC2 is abundantly expressed in goblet cells. To identify the transcriptional activator that regulates goblet-specific expression of MUC2, we analyzed the interaction between the MUC2 promoter and homeodomain proteins CDX1/2, which are involved in the regulation of intestinal development and differentiation. COS-7 cells were transiently transfected with a CDX1 or CDX2 expression construct and then used for the luciferase assay, reverse transcription-polymerase chain reaction, and electrophoretic mobility shift assay (EMSA). The CDX2 expression construct activated the MUC2 promoter and increased the endogenous MUC2 mRNA level, while the CDX1 one did not. EMSA revealed that CDX2 bound to the MUC2 gene cis element, MUC2-WT. These results suggest that CDX2, but not CDX1, interacts with the MUC2 promoter and activates MUC2 transcription, and plays an important role in the differentiation of goblet cells. PMID- 12559946 TI - cDNA cloning and characterization of the third isoform of human peroxin Pex11p. AB - We have cloned a human cDNA encoding an isoform of the peroxin Pex11p, termed Pex11pgamma, by BLASTP homology search on eukaryotic protein database and reverse transcription (RT)-PCR of human fibroblast RNA. Pex11pgamma was 241 amino-acid long, with two putative transmembrane segments, showing 22% and 23% amino-acid identity to Pex11palpha and Pex11pbeta, respectively. PEX11gamma gene was located on chromosome 19, distinct from PEX11alpha and PEX11beta. Pex11pgamma was found to be a peroxisomal membrane protein, as assessed by colocalization with peroxisome targeting signal type 1 (PTS1)-proteins, in epitope-tagged Pex11pgamma expressing Chinese hamster ovary cells. Pex11pgamma exposes both of the N- and C terminal parts to the cytosol. PEX11gamma was not induced in rats by treatment of clofibrate, a peroxisome proliferator, similar to constitutively expressed PEX11beta but in contrast to inducible PEX11alpha. PMID- 12559947 TI - Genomic occurrence of microsatellites containing integral and non-integral repeat numbers. AB - We calculated occurrences of all dinucleotide and trinucleotide microsatellites in the human, mouse, and yeast genomes. The microsatellites were considered separately not only according to the repeated dinucleotide or trinucleotide and the microsatellite length but also according to the starting/terminal nucleotide. The analysis showed that dramatically non-equal amounts occurred in the human genome of microsatellites that differed only by the terminal nucleotides. For example, the 23-mer (TTG)(7)TT occurs 635 times in the human genome whereas (GTT)(7)GT is present only three times in the human genome though the two 23-mers share a 22 nucleotide sequence. The dramatically non-equal occurrences of microsatellites differing only by the terminal nucleotides are observed for most dinucleotide and trinucleotide microsatellites and in all analyzed genomes. We suppose that the strikingly non-equal genomic occurrences of these closely related microsatellites originate from conformational properties of DNA. PMID- 12559948 TI - Butyrate impairs intestinal tumor cell-induced angiogenesis by inhibiting HIF 1alpha nuclear translocation. AB - Butyrate is known to stimulate proliferation of normal crypt cells, whereas it inhibits growth and induces apoptosis in colon cancer cells. We examined the effects of butyrate on colon cancer (Caco-2) cell-induced angiogenesis. HUVEC proliferation was significantly inhibited when incubated with medium conditioned by butyrate-treated Caco-2 cells. Simultaneously, levels of the proangiogenic vascular endothelial growth factor (VEGF) were reduced. HIF-1alpha protein, a transcription factor known to be a key regulator in hypoxia-induced angiogenesis, was upregulated by butyrate. This is in contrast to its importance as a VEGF regulating component. However Western blot of nuclear extracts revealed a downregulation of HIF-1alpha protein. HIF-1alpha DNA-binding activity was also decreased by butyrate. Our findings indicate that HIF-1alpha nuclear sequestration is repressed by butyrate, through inhibition of nuclear translocation. We postulate that diminished HIF-1alpha nuclear presence and activity in butyrate-treated Caco-2 cells could be responsible for decreased VEGF expression and antiangiogenic effects. PMID- 12559949 TI - The gene expression profiling of human visceral adipose tissue and its secretory functions. AB - In order to fully understand the physiological functions of adipose tissue, especially its secretory functions, and to provide a basis for the identification of novel obesity related genes, the gene expression profiling of human visceral adipose tissue was established by using cDNA array. 33P-labelled cDNA, derived from visceral adipose tissue total RNA, was hybridized to a cDNA array containing over 16,000 expressed sequence-tagged clones which represent human singleton genes. The expressed sequence tag (EST) was considered to be expressed in visceral adipose tissue when the ratio of signal to noise was greater than or equal to 2. The results were analyzed with bioinformatics. Totally, 8230 genes were found to be expressed in visceral adipose tissue with 5200 known genes and 3030 known ESTs. Most of 84 secretory proteins, 120 receptors, and 74 transcription factors expressed in adipose tissue were newly identified. Many appetite-regulating related peptides or receptors and some reproduction-related genes were first found to be expressed in adipose tissue. Eight autocrine/paracrine systems were described for the first time in the visceral adipose tissue. These results clearly demonstrate that the visceral adipose tissue has important secretory functions and there is a complex local autocrine/paracrine regulatory network. The present work suggests that the visceral adipose tissue is an important component of the neuroendocrine-immune network and plays an important role in regulating appetite not only via endocrine but also via autocrine/paracrine systems. The visceral adipose tissue might also play a role in regulating reproduction and sexual function. PMID- 12559950 TI - Involvement of heat-shock protein 90 in the interleukin-6-mediated signaling pathway through STAT3. AB - Interleukin-6 (IL-6) is a multifunctional cytokine playing roles in the immune system, hematopoiesis, and acute phase reactions. IL-6 also regulates the growth of various types of human malignant tumors. Here we demonstrate that IL-6-induced gene expression was suppressed by a specific heat-shock protein 90 (Hsp90) inhibitor, geldanamycin (GA) in human hepatoma Hep3B cells. GA also suppressed the IL-6-induced activation of signal transducer and activator of transcription 3 (STAT3) in a human embryonic kidney carcinoma 293T cells. This inhibitory effect of GA on STAT3 activation was reversed by overexpression of Hsp90. Furthermore, Hsp90 directly bound to STAT3 via its N-terminal region, which interacted with GA. We provide evidence that the action of GA on IL-6 functions was due to the inhibition of direct physical interactions between STAT3 and Hsp90, which represents a novel role of Hsp90 in the IL-6 signaling pathways. PMID- 12559951 TI - CpG penta- and hexadeoxyribonucleotides as potent immunomodulatory agents. AB - We demonstrate a new design for immunomodulatory CpG DNA containing two sequences each with as few as five or six-nucleotides joined together via 3(')-3(') linkers. These do not require the -PuPu(Py)CGPyPy- hexameric motif generally found essential for CpG DNA immune stimulation. These novel, short-immunomers show potent immunostimulatory activity manifested by IL-12 and IL-6 secretion in murine spleen cell and PBMC cultures and splenomegaly in vivo. Short-immunomers show strong activation of NF-kappaB and stress-activated signaling pathways and induce cytokines in J774 cell cultures. The same sequences also induce cytokines in healthy human PBMC cultures whereas conventional CpG DNA requires different optimal sequences for murine and human immune cells. Additionally, short immunomers inhibit IL-5 secretion and induce IFN-gamma secretion in conalbumin sensitized mouse spleen cell cultures, suggesting reversal of established Th2 responses to Th1 type responses. Short-immunomer also inhibits growth of MCF-7 human tumor xenograft in nude mice. This is the first report of activity with such short DNA sequences and also of sequences lacking hexameric motifs proposed in earlier studies. PMID- 12559952 TI - Characterization of KIBRA, a novel WW domain-containing protein. AB - In a yeast two hybrid screen with the human isoform of Dendrin (KIAA0749), a putative modulator of the postsynaptic cytoskeleton, we isolated a cDNA coding for a novel protein, KIBRA, possessing two amino-terminal WW domains, an internal C2-like domain and a carboxy-terminal glutamic acid-rich stretch. Northern blot analysis revealed that the expression of KIBRA mRNA was predominately found in kidney and brain. In vitro interaction studies revealed that the first KIBRA WW domain binds specifically to PPxY motifs. Transient transfection of monkey kidney cells with constructs encoding Myc-tagged KIBRA displayed a cytoplasmic localization and a perinuclear enrichment of the protein. PMID- 12559953 TI - Molecular cloning and mRNA expression analysis of a novel rice (Oryzasativa L.) MAPK kinase kinase, OsEDR1, an ortholog of Arabidopsis AtEDR1, reveal its role in defense/stress signalling pathways and development. AB - Mitogen-activated protein kinase (MAPK) cascade(s) is important for plant defense/stress responses. Though MAPKs have been identified and characterized in rice (Oryza sativa L.), a monocot cereal crop research model, the first upstream component of the kinase cascade, namely MAPK kinase kinase (MAPKKK) has not yet been identified. Here we report the cloning of a novel rice gene encoding a MAPKKK, OsEDR1, designated based on its homology with the Arabidopsis MAPKKK, AtEDR1. OsEDR1, a single copy gene in the genome of rice, encodes a predicted protein with molecular mass of 113046.13 and a pI of 9.03. Using our established two-week-old rice seedling in vitro model system, we show that OsEDR1 has a constitutive expression in seedling leaves and is further up-regulated within 15 min upon wounding by cut, treatment with the global signals jasmonic acid (JA), salicylic acid (SA), ethylene (ethephon, ET), abscisic acid, and hydrogen peroxide. In addition, protein phosphatase inhibitors, fungal elicitor chitosan, drought, high salt and sugar, and heavy metals also dramatically induce its expression. Moreover, OsEDR1 expression was altered by co-application of JA, SA, and ET, and required de novo synthesized protein factor(s) in its transient regulation. Furthermore, using an in vivo system we also show that OsEDR1 responds to changes in temperature and environmental pollutants-ozone and sulfur dioxide. Finally, OsEDR1 expression varied significantly in vegetative and reproductive tissues. These results suggest a role for OsEDR1 in defense/stress signalling pathways and development. PMID- 12559954 TI - BpV (phen) induces apoptosis of RINm5F cells by modulation of MAPKs and MKP-1. AB - We investigated the mechanism of toxicity of peroxovanadium complex bpV (phen) in RINm5F cells. Treatment with bpV (phen) provoked cell death, predominantly by apoptosis. This compound induced strong and sustained JNK and p38 MAPK activation. However, ERK phosphorylation was not affected. The level of expression of MAPK phosphatase MKP-1 was suppressed after bpV (phen) treatment. In addition, this compound did not stimulate proteolytic processing of procaspase 3, suggesting that caspase-3 is not activated during the course of bpV (phen) induced apoptosis. A correlative inhibition of JNK activation by immunosuppressive drug FK 506 induced ERK activation and MKP-1 expression, and suppressed RINm5F cell death. A specific p38 inhibitor SB 203580 also stimulated ERK activation and cell survival. Furthermore, simultaneous pretreatment with both FK 506 and SB 203580 almost completely abolished cell death. Thus, our results suggest that stress kinases and MKP-1 have a role in bpV (phen)-induced apoptosis of RINm5F cells. PMID- 12559955 TI - Cyclooxygenase 2 inhibits SAPK activation in neuronal apoptosis. AB - Cyclooxygenase 2 (COX-2) expressed in cultured neuronal PC12 cells under inducible promoter protects cells from trophic withdrawal apoptosis. Stimulation of SAPK is thought to play a significant role in initiation of PC12 cell death. We have therefore examined whether COX-2 expression inhibits trophic withdrawal mediated activation of SAPK. SAPK activity increased during the first 6h after NGF removal in mock-transfected PC12 cells. COX-2 expression attenuated the increase of SAPK, as detected by Western blot analysis with phosphorylation state specific anti-SAPK antibodies and by SAPK activity assays. We propose that COX-2 attenuated SAPK activation by preventing activation of nNOS, which occurs, as we have shown before, via COX-2-mediated expression of dynein light chain (DLC). Activation of SAPK in neuronal cell death was attenuated by DLC expression. These observations support a role for NO production and SAPK activation in the neuronal death mechanisms. PMID- 12559956 TI - Calcium-dependent translocation of cytosolic phospholipase A2 in pancreatic beta cells. AB - Cytosolic phospholipase A(2)(cPLA(2)), an enzyme responsible for the generation of arachidonic acid, is located in the cytosolic compartment in most tissues and it translocates to membrane compartments when activated. We found that cPLA(2) distribution in pancreatic beta-cells is different from that of most other mammalian cells: it is evenly distributed throughout the beta-cell, in both cytoplasmic and nuclear compartments. Agents that increased intracellular Ca(2+) in the MIN6 beta-cell line also stimulated a redistribution of cPLA(2) immunoreactivity such that the majority of the enzyme moved from the nucleus to the cytoplasm. The time course of events was compatible with the elevation in Ca(2+) being responsible for translocation of cPLA(2). These observations suggest that cPLA(2) may be compartmentalised in unstimulated beta-cells, perhaps to limit its access to substrate prior to elevations in intracellular Ca(2+). PMID- 12559957 TI - Male specific expression of a cytochrome P450 (Cyp312a1) in Drosophila melanogaster. AB - Using cDNA array techniques, the transcriptional levels of Drosophila cytochrome P450 (P450) genes were compared between male and female flies. Of the 86 P450s, 15 isoforms were picked up and the levels of transcription were confirmed by the real time quantitative RT-PCR. Cyp315a1 and Cyp302a1, which had been reported as P450s involved in the ecdysteroid biosynthesis, were included in the 15 isoforms and expression of these P450s was 8.6- and 7.9-fold higher in females than in males, respectively. In addition, we confirmed that expression of Cyp312a1 was 82 fold higher in adult males than females. This gene expression was observed mostly in the abdomen and its transcription level gradually increased from pupal stage and peaked in the 5-day-old adult. Furthermore, the male specific expression of Cyp312a1 was universally observed in three Drosophila strains originated from the USA (Oregon R), China (Canton S), and Japan (HKJ), suggesting possible involvement of this P450 in significant endogenous catalytic reaction(s). This is the first report of a P450 enzyme being predominantly expressed in male Drosophila. PMID- 12559958 TI - Shortening of microsatellite deoxy(CA) repeats involved in GL331-induced down regulation of matrix metalloproteinase-9 gene expression. AB - Matrix metalloproteinase-9 (MMP-9) associates with cancer cell invasion and metastasis. CL1-5 cells, a human lung adenocarcinoma cell line, expressed an elevated level of MMP-9 and exhibited a highly invasive and metastatic ability. By Matrigel assay and gelatinase zymography, the topoisomerase II poison GL331 was found to dose-dependently inhibit the invasiveness and the level of secreted MMP-9 of CL1-5 cells. Northern blot analysis indicated that cellular MMP-9 mRNA level was decreased after GL331 treatment. Furthermore, GL331-induced down regulation of mmp-9 gene promoter was demonstrated by using a luciferase reporter gene driven by the -216 to -13 region of the mmp-9 gene promoter cloned from CL1 5 cells. By PCR amplification and gel electrophoresis, we found that GL331 caused shortening of the -216 to -13 region of the mmp-9 promoter. Direct sequencing analysis revealed that the number of d(CA) was reduced from 24 to 18 at the microsatellite d(CA) repeat region of the mmp-9 promoter. The CL1-5 cells transfected with the luciferase reporter containing 18 d(CA)s expressed only 53% of those when the reporter contained 24 d(CA)s. The promoter region of mmp-9 gene contains other positive regulatory elements, such as TRE and kappaB. We found that GL331 did not significantly influence the luciferase activity driven by TRE or kappaB. Taken together, these data suggested that GL331 inhibited MMP-9 mRNA expression at least partly through the selective induction of shortening of microsatellite d(CA) repeats. This is the first report that an anti-cancer agent can inhibit mmp-9 gene expression by inducing microsatellite DNA shortening. PMID- 12559959 TI - OTX2 regulates expression of DOPAchrome tautomerase in human retinal pigment epithelium. AB - Otx2 is a member of homeodomain-containing transcription factors and is essential for eye morphogenesis in mice. Here we show the expression of OTX2, the human counterpart of Otx2, in cell lines of retinal pigment epithelium (RPE) and in Y79 retinoblastoma cells that exhibit the property of presumptive RPE. These RPE cells express DOPAchrome tautomerase (DCT) that is an enzyme involved in melanin biosynthesis. DCT may contribute to the homeostasis of RPE by detoxifying DOPA derived metabolites. OTX2 binds to the DCT gene promoter in vivo, as judged by chromatin immunoprecipitation assays. Furthermore, repression of endogenous OTX2 expression in Y79 cells by an anti-sense OTX2 oligonucleotide resulted in the decrease of DCT protein contents. Transient expression assays revealed that OTX2 activated the DCT gene promoter through the OTX-2-binding site in an RPE-specific manner. Therefore, OTX2 may regulate RPE-specific target genes, such as DCT, thereby maintaining the homeostasis of RPE. PMID- 12559960 TI - A faithful method for PCR-mediated global mRNA amplification and its integration into microarray analysis on laser-captured cells. AB - Quantitative and qualitative analyses of mRNAs from a small number of cells are extremely important for studies on gene expression in various physiological and pathological conditions in multicellular organisms. We present here an effective method for high-fidelity global mRNA amplification for in vivo gene expression profiling of as few as 100 cells obtained by laser-captured microdissection (LCM). This method, called TALPAT, is based on T7 RNA polymerase-mediated transcription, adaptor ligation, and PCR amplification followed by T7 transcription. More than 80% of genes were commonly identified as a more than 3 fold changed gene among three gastric cancer cell lines using cRNA amplified by both TALPAT and the ordinary in vitro T7-transcription. The reproducibility of TALPAT was validated by microarray analysis on 100 breast cancer cells obtained by LCM. For the application of the LCM-TALPAT method, we successfully obtained expression profiles of gastric cancer cells and the mesenchymal cells, enabling us to understand in vivo cell-to-cell cross-talk in the microenvironment. PMID- 12559961 TI - Oligomerization characteristics of cysteine string protein. AB - CSP function is vital to synaptic transmission, however; the precise nature of its role remains controversial. Conflicting reports support either a role for CSP: (i) in exocytosis or (ii) in the regulation of transmembrane calcium fluxes. Here we have examined the self-association of CSP to form oligomers that are stable upon SDS-PAGE. To understand the structural requirements for CSP self association a series of CSP deletion mutants were constructed, expressed, and purified. This analysis revealed an interesting pattern of oligomerization. Amino acids between 83 and 136 were observed to be important for self-association. The recombinant CSP oligomers as well as the CSP monomers directly associate with Ni(2+)-NTA agarose. Thus CSP-CSP interactions may be an important consideration for current working models of CSP chaperone activity at the synapse. PMID- 12559962 TI - Plexin-A1 and plexin-B1 specifically interact at their cytoplasmic domains. AB - Semaphorin 3A (Sema3A) is a member of semaphorins and functions as an axonal repulsive guidance molecule. Neuropilin-1 and plexin-As form receptor complexes for Sema3A and plexin-As are thought to initiate the intracellular signaling cascade. However, the molecule by which plexin-As transduce their signal is not well understood. We searched molecules that interact with intracellular domains of plexin-A1 by yeast two-hybrid screening and identified a 349 amino acid fragment of plexin-B1 as a plexin-A1 interacting protein. We, then, cloned mouse plexin-B1 and confirmed their interaction in a mammalian expression system. Plexin-B1 physically associated with plexin-A1, but not with plexin-A2 or A3. Northern blot analysis showed the expression of both plexin-A1 and B1 in adult brain. We propose that plexin-A1 and B1 interact in the adult brain and transduce Sema3A signaling in cooperation. PMID- 12559963 TI - Chronic methamphetamine administration reduces histamine-stimulated phosphoinositide hydrolysis in mouse frontal cortex. AB - In the present study, it was hypothesized that in vivo pretreatment with repeated methamphetamine would alter the agonist-stimulated phosphoinositide hydrolysis in mouse frontal cortical slices. Male ICR mice that received the methamphetamine injection (1.0mg/kg, intraperitoneally) once a day for five consecutive days showed behavioral sensitization to the same dose of methamphetamine 5 days after the last injection of the initial chronic treatment regimen (test day 10). On test day 10, the reduction of histamine (0.1-1.0mM)-stimulated phosphoinositide hydrolysis in the mouse frontal cortex was observed. The reduction was specific to histamine, but not to norepinephrine (10 microM-0.1mM) or L-glutamate (0.1 0.5mM). The reduction occurred without any change in the expression level of histamine H(1) receptor mRNA. The reduction recovered 25 days after the last injection of the initial chronic treatment regimen (test day 30). The direct application to the slices of a pharmacologically effective concentration of methamphetamine in vitro (10 microM) did not alter the histamine signal transduction. The present results suggest that the reduction is probably one of neuroadaptations in the frontal cortex contributing to behavioral sensitization. PMID- 12559964 TI - P27 knockout mice: reduced myostatin in muscle and altered adipogenesis. AB - Knockout of the P27(kip) gene, which encodes a cyclin-dependent kinase inhibitor involved in cell proliferation regulation, results in growth enhancement in mice. To investigate how p27 deficiency affected adipogenesis and myogenesis, levels of PPARgamma, C/EBPalpha, and the myogenesis inhibitor, myostatin, were measured in p27(-/-) (n=14), p27(+/-) (n=18), and p27(+/+) mice (n=11). Body weight and gastrocnemius muscle (GC) mass were increased in p27(-/-) mice (P<0.05), but there were no differences in fat depot weights, percent body fat or serum leptin concentrations among genotypes. PPARgamma, but not C/EBPalpha, was markedly increased in p27(-/-) mice (P<0.05). There was also a higher incidence of inguinal fat apoptosis (P<0.01) in p27(-/-) mice. Myostatin levels were reduced in GC muscle of p27(-/-) mice (P<0.05). These findings suggest that in p27 deficient mice, increased skeletal muscle mass is mediated in part through decreased myostatin. Although total adiposity was not changed, increased PPARgamma levels suggest an alteration in adipogenesis. PMID- 12559965 TI - Aryl hydrocarbon receptor ligands repress T-cadherin expression in vascular smooth muscle cells. AB - T-cadherin, a glycosylphosphatidylinositol-modified cadherin subtype, is highly expressed in cardiac and vascular tissues. Neither the functions nor regulation of T-cadherin in these tissues is understood. We have cloned rat T-cadherin cDNA encoding the full length amino acid sequence. The 5(') untranslated nucleotide sequences of rat, mouse, and human T-cadherin contain a conserved GCGTG motif which constitutes the invariant core sequence of dioxin- or xenobiotic-regulatory elements. These elements function as target sites for aryl hydrocarbon receptor/aryl hydrocarbon nuclear translocator (AhR/ARNT) in genes regulated by this transcription factor. Using cultures of rat aortic smooth muscle cells this study presents data revealing T-cadherin as a putative target gene for negative regulation of expression through AHR signalling. Prototypic AHR agonists benzo[a]pyrene (BaP) or 7,12-dimethylbenzanthracene (DMBA) and 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) repressed T-cadherin mRNA levels. Repression was antagonized by the cognate AHR antagonist alpha-naphthoflavone (alpha-NF). Repression was insensitive to inhibitors of gene transcription (actinomycin D) or de novo protein synthesis (cycloheximide), suggesting AHR/ARNT functions directly in transcriptional repression of T-cad. Regulation of adhesion proteins through the AHR pathway may represent a novel mechanism of action by atherogenic polycyclic aromatic hydrocarbons. PMID- 12559966 TI - Cleavage of nucleolin and argyrophilic nucleolar organizer region associated proteins in apoptosis-induced cells. AB - To investigate the behavior of nuclear proteins in apoptotic cells, we examined the changes in nucleolin and proteins of the nucleolar organizing region during apoptosis in human osteoblastic cell lines, Saos-2 and MG63. Apoptosis was induced by treatment of these cells with okadaic acid. Proteins prepared from apoptotic cells were subjected to Western blot analysis and a modified Western blot method using silver nitrate. The anti-nucleolin antibody recognized the 110 kDa band and the staining intensity of this band decreased in the proteins prepared from the okadaic acid-treated apoptotic cells. The additional band of an 80-kDa was also detected in the proteins prepared from the apoptotic cells. Two major silver nitrate-stained bands, 110-kDa and 37-kDa, were detected among the proteins obtained from control cells. Like the Western blot analysis, the intensity of the 110-kDa silver nitrate-staining band decreased; an 80-kDa band appeared and its staining intensity increased in the lysate from the okadaic acid treated cells. The signal intensity of the 37-kDa protein did not change in the sample from the apoptotic cells. In a cell-free apoptotic system, the 80-kDa protein was also detected and the amount of the 110-kDa protein decreased in the extract of Saos-2 cell nuclei incubated with apoptotic cytosol. The change in nucleolin in Saos-2 cells induced to undergo apoptosis was examined by an immunocytochemical procedure using the anti-nucleolin antibody and Hoechst 33342. Nucleolin was visible as dots in nucleoli in the control cells; however, it was not detected in the cells undergoing apoptosis. The dual-exposure view of Hoechst 33342 and anti-nucleolin staining cells confirmed that nucleolin had disappeared from the apoptotic nuclei of Saos-2. PMID- 12559967 TI - Role of prostaglandin E produced by osteoblasts in osteolysis due to bone metastasis. AB - Prostaglandin E2 (PGE2) is produced in bone mainly by osteoblasts and stimulates bone resorption. Osteolytic bone metastasis of cancers is accompanied by bone resorption. In this study, we examined the roles of PGE2 in osteolysis due to bone metastasis of breast cancer. Injection of human breast cancer cells, MDA-MB 231 (MDA-231), into nude mice causes severe osteolysis in the femur and tibia. The expression of cyclo-oxygenase-2 (COX-2) and the receptor activator of NF kappaB ligand (RANKL), a key molecule in osteoclast differentiation, mRNAs was markedly elevated in bone with metastasis. When MDA-231 cells were cocultured with mouse calvaria, COX-2-induced PGE2 production and bone resorption progressed. The contact with MDA-231 cells could induce the expression of COX-2 and RANKL in osteoblasts by mechanisms involving MAP kinase and NF-kappaB. The blockage of PGE2 signal by indomethacin and EP4 antagonist abrogated the osteoclast formation induced by the breast cancer cells. Here, we show a PGE dependent mechanism of osteolysis due to bone metastasis. PMID- 12559968 TI - Inhibition of myostatin in adult mice increases skeletal muscle mass and strength. AB - A human therapeutic that specifically modulates skeletal muscle growth would potentially provide a benefit for a variety of conditions including sarcopenia, cachexia, and muscular dystrophy. Myostatin, a member of the TGF-beta family of growth factors, is a known negative regulator of muscle mass, as mice lacking the myostatin gene have increased muscle mass. Thus, an inhibitor of myostatin may be useful therapeutically as an anabolic agent for muscle. However, since myostatin is expressed in both developing and adult muscles, it is not clear whether it regulates muscle mass during development or in adults. In order to test the hypothesis that myostatin regulates muscle mass in adults, we generated an inhibitory antibody to myostatin and administered it to adult mice. Here we show that mice treated pharmacologically with an antibody to myostatin have increased skeletal muscle mass and increased grip strength. These data show for the first time that myostatin acts postnatally as a negative regulator of skeletal muscle growth and suggest that myostatin inhibitors could provide a therapeutic benefit in diseases for which muscle mass is limiting. PMID- 12559970 TI - TGFbeta down-regulates IFN-gamma production in IL-18 treated NK cell line LNK5E6. AB - Transforming growth factor beta (TGFbeta) is a critical immunosuppressive cytokine that inhibits the cell-mediated immune responses partly via inhibition of immunostimulatory cytokine production from T cells, NK cells, and macrophages. Here we investigated the effect of TGFbeta on NK cell activation induced by interleukin 18 (IL-18) using a murine NK cell line LNK5E6. IL-18 activated LNK5E6 cells to produce antiviral activity against vesicular stomatitis virus (VSV) and TGFbeta inhibited this activation. TGFbeta inhibited interferon-gamma (IFN-gamma) production in LNK5E6 cells treated with IL-18. TGFbeta also suppressed the IL-18 induced mRNA expression of IFN-gamma. Moreover, TGFbeta did not affect the transcriptional activity of IFN-gamma but decreased the half-life of IFN-gamma mRNA induced by IL-18. These results suggest that the destabilization of IFN gamma mRNA induced by TGFbeta leads to the inhibition of antiviral activity and IFN-gamma production in IL-18 stimulated LNK5E6 cells. PMID- 12559969 TI - Molecular cloning and characterization of the human ASB-8 gene encoding a novel member of ankyrin repeat and SOCS box containing protein family. AB - We have cloned a new member of human ankyrin repeat and SOCS box containing protein family (ASB), designed as hASB-8, from a human placental cDNA library and further extended by 5(') and 3(')-RACE. The full-length cDNA was 2545bp in length, with a predicted open reading frame encoding a protein of 288 amino acids, which was 96% identical to mouse ASB-8 protein. Computer analysis revealed that the deduced amino acid sequence of the human ASB-8 contained four Ankyrin repeats and one SOCS box. The gene had four exons separated by three introns and was mapped to human chromosome 12q13. Human ASB-8 mRNA was expressed at the highest level of expression in skeletal muscle and at a varied level of expression in heart, brain, placenta, liver, kidney, and pancreas. The transcript of hASB-8 was not detected in adult normal lung tissue, but found in lung carcinoma cell lines SPC-A1, A549, and NCI-H446. Subcellular localization analysis showed that the EGFP-tagged hASB-8 protein was localized at cytoplasm in human hepatocellular carcinoma cell line BEL-7402. We also provided evidence that hASB-8 could interact with Elongin B-C complex in vitro. Furthermore, transfection with the truncated mutant of hASB-8 cDNA lacking SOCS box could suppress cell growth of lung adenocarcinoma SPC-A1 cells in vitro, which suggests that this gene might be related to the development of lung cancer. PMID- 12559971 TI - In vitro effects of polyglutamine tracts on Ca2+-dependent depolarization of rat and human mitochondria: relevance to Huntington's disease. AB - The mechanisms by which neurons die in CAG triplet repeat (polyglutamine) disorders, such as Huntington's disease, are uncertain; however, mitochondrial dysfunction and disordered calcium homeostasis have been implicated. We previously demonstrated abnormal mitochondrial calcium handling in Huntington's disease cell lines and transgenic mice. To examine whether these abnormalities might arise in part from direct effects of the expanded polyglutamine tract contained in mutant huntingtin, we have exposed normal rat liver and human lymphoblast mitochondria to glutathione S-transferase fusion proteins containing polyglutamine tracts of 0, 19, or 62 residues. Similar to bovine serum albumin, each of the protein constructs nonspecifically inhibited succinate-supported respiration, independent of polyglutamine tract length. There was a small but significant reduction of mitochondrial membrane potential (state 4) only in the presence of the pathological-length polyglutamine tract. With successive addition of small Ca(2+) aliquots, mitochondria exposed to pathological-length polyglutamine tracts (approximately 5 microM) depolarized much earlier and to a greater extent than those exposed to the other protein constructs. These results suggest that the mitochondrial calcium handling defects seen in Huntington's disease cell lines and transgenic mice may be due, in part, to direct, deleterious effects of mutant huntingtin on mitochondria. PMID- 12559972 TI - Mechanism of Janus kinase 3-catalyzed phosphorylation of a Janus kinase 1 activation loop peptide. AB - A high-affinity IL-2 receptor requires two Janus protein tyrosine kinases (JAKs) for IL-2 signal transduction: JAK1 and JAK3. Since transphosphorylation of the two kinases is presumed to occur after receptor engagement we examined the phosphorylation by recombinant JAK3 of a peptide substrate corresponding to the JAK1 activation loop (KAIETDKEYYTVKD), which has two adjacent tyrosines. Mass spectral analysis of the enzymatically phosphorylated peptide showed that the second tyrosine was phosphorylated at a 30-fold greater rate than the first tyrosine. Moreover, no doubly phosphorylated peptide was detected by this analysis. Kinetic analysis of the reactions of singly phosphorylated JAK1 activation loop peptides showed that phosphorylating the first or second tyrosine decreased the k(cat)/K(m) for the phosphorylation of the other 115- and 26-fold, respectively. Singly changing each side chain of the KEYYTV portion of the peptide to a methyl group (alanine) yielded substrates comparable to the wild type sequences in all cases except that of the first or second tyrosine, which showed a 153- or 70-fold drop in k(cat)/K(m), respectively. Using libraries of immobilized peptides with all 20 naturally occurring amino acids substituted for Y9 or T11 showed that the JAK3 tolerated substitution at T11 but prefers large hydrophobic amino acids at Y9. These results show that JAK3 does not act processively on the JAK1 activation loop in vitro and illustrate the role of Y9 in the recognition of the preferred site of phosphorylation which is Y10. PMID- 12559973 TI - Activation of cytochrome P450 2C9-mediated metabolism: mechanistic evidence in support of kinetic observations. AB - Studies were designed to investigate the possible mechanisms associated with the kinetic observation of CYP2C9 activation by dapsone and its phase I metabolite, N hydroxydapsone. Kinetic studies suggested that dapsone activated CYP2C9-mediated flurbiprofen 4(')-hydroxylation by decreasing the K(m) (alpha=0.2) and increasing the V(max) (beta=1.9). Interestingly, N-hydroxydapsone also activated flurbiprofen 4(')-hydroxylation by increasing V(max) (beta=1.5) but had no effect on K(m) (alpha=0.98). To study the effects of these modulators on the binding affinity of flurbiprofen, spectral binding studies were performed. In the presence of dapsone, the spectral binding constant (K(s)) for flurbiprofen was reduced from 14.1 to 2.1 microM, while in the presence of N-hydroxydapsone, the K(s) remained unchanged (14.0 microM), which suggests that dapsone causes an increase in the affinity of flurbiprofen for CYP2C9, whereas N-hydroxydapsone does not. Additionally, stoichiometry measurements under activation conditions in the presence of dapsone resulted in a doubling of both NADPH and oxygen consumption for flurbiprofen 4(')-hydroxylation, with an overall increase in metabolite formation and a decrease in formation of peroxide and excess water. Interestingly, the presence of N-hydroxydapsone generally caused the same effects on stoichiometry as those of flurbiprofen 4(')-hydroxylation but failed to reduce excess water formation, which suggests that, while N-hydroxydapsone activates CYP2C9, it does so less efficiently and possibly through a mechanism different from that of dapsone. PMID- 12559974 TI - Force maintenance in smooth muscle: analysis using sinusoidal perturbations. AB - The "latch state" or force maintenance may be due to the emergence of a distinct set of dephosphorylated, slowly cycling "latch" cross-bridges, slowing of the overall cross-bridge cycling rate, or a non-cross-bridge contribution. This was investigated by sinusoidally oscillating strips of intact rabbit portal vein or aorta. Tissue strips were activated with KCl depolarization, resulting in a sustained increase of MLC(20) phosphorylation or 10 microM phenylephrine, resulting in a transient increase in MLC(20) phosphorylation. Stiffness was calculated from the force response to a small, sine-wave oscillation in muscle length (1-100 Hz). The results produced a 3-dimensional plot of stiffness versus the frequency of oscillation (Hz) versus time (s), or stiffness distribution profile. During KCl depolarization, the stiffness distribution profile displayed a shift toward lower frequencies, suggesting a general slowing in the overall cross-bridge cycling rate during force maintenance. On the other hand, phenylephrine stimulation did not display a significant change in the stiffness distribution profile, suggesting that the overall cross-bridge cycling rate did not significantly change during force maintenance. PMID- 12559975 TI - Rapid regulation of collagen but not metalloproteinase 1, 3, 13, 14 and tissue inhibitor of metalloproteinase 1, 2, 3 expression in response to mechanical loading of cartilage explants in vitro. AB - This study analyzes the molecular response of articular chondrocytes to short term mechanical loading with a special focus on gene expression of molecules relevant for matrix turnover. Porcine cartilage explants were exposed to static and dynamic unconfined compression and viability of chondrocytes was assessed to define physiologic loading conditions. Cell death in the superficial layer correlated with mechanical loading and occurred at peak stresses >or=6 MPa and a cartilage compression above 45%. Chondrocytes in native cartilage matrix responded to dynamic loading by rapid and highly specific suppression of collagen expression. mRNA levels dropped 11-fold (collagen 2; 6 MPa, P=0.009) or 14-fold (collagen 1; 3 and 6 MPa, P=0.009) while levels of aggrecan, tenascin-c, matrix metalloproteinases (MMP1, 3, 13, 14), and their inhibitors (TIMP1-3) did not change significantly. Thus, dynamic mechanical loading rapidly shifted the balance between collagen and aggrecan/tenascin/MMP/TIMP expression. A better knowledge of the chondrocyte response to mechanical stress may improve our understanding of mechanically induced osteoarthrits. PMID- 12559976 TI - Association of protein-DNA recognition complexes: electrostatic and nonelectrostatic effects. AB - In this study the electrostatic and nonelectrostatic contributions to the binding free energy of a number of different protein-DNA recognition complexes are investigated. To determine the electrostatic effects in the protein-DNA association the Poisson-Boltzmann approach was applied. Overall the salt dependent electrostatic free energy opposed binding in all protein-DNA complexes except one, and the salt-independent electrostatic contribution favored binding in more than half of the complexes. Further the salt-dependent electrostatic free energy increased with higher ionic concentrations and therefore complex association is stronger opposed at higher ionic concentrations. The hydrophobic effect in the protein-DNA complexes was determined from the buried accessible surface area and the surface tension. A majority of the complexes showed more polar than nonpolar buried accessible surface area. Interestingly the buried DNA accessible surface area was preferentially hydrophilic, only in one complex a slightly more hydrophobic buried accessible surface area was observed. A quite sophisticated balance between several different free energy components seems to be responsible for determining the free energy of binding in protein-DNA systems. PMID- 12559977 TI - Peptide scanning-based identification of regions of gamma-II crystallin involved in thermal aggregation: evidence of the involvement of structurally analogous, helix-containing loops from the two double Greek key domains of the molecule. AB - Gamma crystallin is one of three structural proteins present in great abundance in the fiber cells of the vertebrate eye lens. The protein displays a tendency to aggregate readily in the course of heating, cooling, being exposed to ultraviolet radiation, or rapid refolding. To investigate the molecular mechanisms underlying such aggregation, we have employed a peptide-scanning approach aimed at identifying regions of bovine gamma-II crystallin that may be involved in intermolecular interactions leading to aggregation, using assays that measure the competitive inhibition of such aggregation by reagents drawn from a group of contiguous (overlapping) peptides derived from the sequence of the protein itself. Our results suggest that two regions, comprising residues 61-74, and 145 159, play key roles in aggregative interactions. Intriguingly, the two regions (each containing a solvent-exposed, single-turn helix in the native structure) are located in structurally analogous positions in the two homologous double Greek key (beta sheet) domains of the protein, suggesting that helix-strand conversions may operate to facilitate intermolecular beta sheet interactions during aggregation. PMID- 12559978 TI - Effect of different metal ions on the oxidative damage and antioxidant capacity of hyaluronic acid. AB - Degradation and the antioxidative effect of Na-, Zn-, Co-, Cu-, and Mn-hyaluronic acid (HA) associates were studied. Our findings revealed the protective effect of certain counterions against ROS-induced HA degradation. We could also separate the antioxidative effect of certain counterions from that of the HA by examining the effect of the counterions in their free ionic forms. The result showed that metal ions with altering oxidative status (Co(2+), Cu(2+), Mn(2+)) proved to be effective in themselves or their effect added to that of HA when HA was also effective. Moreover, the effects of Co-HA against z.rad;O(2)(-) and of Mn-HA against ONOO(-) as well as the synergic effect of Zn-HA associates where Zn(2+) is of fixed oxidative status were attributed to the structure-stabilizing complex formed between certain counterions and HA. Our examination also concerned the influence of HA associates on the indirect antioxidation related to Fe(2+) chelating. The individual effects of Zn(2+), Co(2+), and Cu(2+) were only detectable, which could be explained by the competitive displacement of ferrous from its binding site. PMID- 12559979 TI - Thiol-activated serine proteinases from nymphal hemolymph of the African migratory locust, Locusta migratoria migratorioides. AB - Two unique serine proteinase isoenzymes (LmHP-1 and LmHP-2) were isolated from the hemolymph of African migratory locust (Locusta migratoria migratorioides) nymphs. Both have a molecular mass of about 23 kDa and are activated by thiol reducing agents. PMSF abolishes enzymes activity only after thiol activation, while the cysteine proteinase inhibitors E-64, iodoacetamide, and heavy metals fail to inhibit the thiol-activated enzymes. The N-terminal sequence was determined for the more-abundant LmHP-2 isoenzyme. It exhibits partial homology to that of other insect serine proteinases and similar substrate specificity and inhibition by the synthetic and protein trypsin inhibitors pABA, TLCK, BBI, and STI. The locust trypsins LmHP-1 and LmHP-2 constitute a new category of serine proteases wherein the active site of the enzyme is exposed by thiol activation without cleavage of peptide bonds. PMID- 12559980 TI - Nitroxyl-mediated disruption of thiol proteins: inhibition of the yeast transcription factor Ace1. AB - Among the biologically and pharmacologically relevant nitrogen oxides, nitroxyl (HNO) remains one of the most poorly studied and least understood. Several previous reports indicate that thiols may be a primary target for the biological actions of HNO. However, the intimate details of the chemical interaction of HNO with biological thiols remain unestablished. Due to their ability to grow under a variety of conditions, the yeast Saccharomyces cerevisiae represents a unique and useful model system for examining the chemistry of HNO with thiol proteins in a whole-cell preparation. Herein, we have examined the effect of HNO on the thiol containing, metal-responsive, yeast transcription factor Ace1 under a variety of cellular conditions as a means of delineating the chemistry of HNO interactions with this representative thiol protein. Using a reporter gene system, we find that HNO efficiently inhibits copper-dependent Ace1 activity. Moreover, this inhibition appears to be a result of a direct interaction between Ace1 thiols and HNO and not a result of any chemistry associated with HNO-derived species. Thus, this report indicates that thiol proteins can be a primary target of HNO biochemistry and that HNO-mediated thiol modification is likely due to a direct reaction of HNO. PMID- 12559981 TI - Establishing the inhibitory effects of bradykinin on thrombin. AB - Bradykinin, RPPGFSPFG, has been reported to be an inhibitor of thrombin's roles in blood clotting, platelet activation, and cellular permeability. The exact target, magnitude, and type of inhibition occurring are not well characterized. Based on the individual kinetic parameters calculated here, bradykinin is classified as a weak competitive inhibitor against hydrolysis of S-2238 and of a PAR4-like peptide. The K(m) values increased twofold in the presence of bradykinin, whereas the k(cat) values remained constant. The K(i) values ranged from 170 to 326 microM. Other biochemical studies indicated that bradykinin inhibits release of fibrinopeptide A from fibrinogen. Furthermore, bradykinin hindered the time required for fibrin clot formation. The weak inhibitions observed in vitro suggest that the direct effects of bradykinin on the thrombin active site become significant only at high concentrations, levels that may be difficult to achieve physiologically. Clearly, bradykinin can target thrombin but whether this direct interaction can be achieved in vivo and is sufficient to elicit a response without contributions from other cofactors requires further investigation. PMID- 12559982 TI - [14C]serine from phosphatidylserine labels ceramide and sphingomyelin in L929 cells: evidence for a new metabolic relationship between glycerophospholipids and sphingolipids. AB - After incubation of L929 cells with [14C]serine and various effectors an inverse correlation between label in ceramide and phosphatidylserine (PS) was displayed. This surprising behavior of the two metabolites prompted us to check whether serine of PS could be a source for ceramide synthesis. We therefore incubated L929 cells for 30 min in serum-free medium with L-phosphatidyl-L-[3-14C]serine in the presence or in the absence of cycloserine, an established inhibitor of serine palmitoyltransferase. During this short period L-phosphatidyl-L-[3-14C]serine labeled ceramide and this label was suppressed by cycloserine. Then L929 cells were grown for 16-18 h in the presence of L-phosphatidyl-L-[3-14C]serine. After this period the label was seen in sphingomyelin. Labeling of ceramide and sphingomyelin by serine from PS provides evidence for a new metabolic relationship between glycerophospholipids and sphingolipids. PMID- 12559983 TI - Age-related decline of sodium-dependent ascorbic acid transport in isolated rat hepatocytes. AB - This study investigated whether the age-related decline in hepatic ascorbic acid (AA) levels in rats was due to altered AA uptake. AA concentrations were 68% lower in freshly isolated hepatocytes from old (24-26 months) versus young (3-5 months; p<0.0005) Fischer 344 rats. When incubated with 100 microM AA, cells from old as compared to young rats showed a 66% decline in both the rate of AA transport and the steady state intracellular levels. Sodium-free media significantly reduced AA uptake, suggesting that the sodium-dependent vitamin C transporter (SVCT) was largely responsible for declines in AA transport. Analysis of SVCT messenger RNA (mRNA) levels shows that one isoform of this transport protein, SVCT1, declines 45% with age, with no significant changes in SVCT2 mRNA levels. These results show for the first time that sodium-dependent AA transport declines during the aging process, which may account for much of the loss in tissue AA content. PMID- 12559984 TI - The interaction of sodium chlorite with phospholipids and glutathione: a comparison of effects in vitro, in mammalian and in microbial cells. AB - In this study the interaction of the preservative sodium chlorite with unsaturated lipids and glutathione was investigated, in comparison with peroxides, sodium hypochlorite, and benzalkonium chloride. The aim was to determine whether the action of sodium chlorite could involve membrane lipid damage or antioxidant depletion, and how this related to toxicity in both mammalian and microbial cells. The treatment of phospholipids with chlorite yielded low levels of hydroperoxides, but sodium chlorite oxidized the thiol containing antioxidant glutathione to its disulfide form very readily in vitro, with a 1:4 oxidant:GSH stoichiometry. In cultured cells, sodium chlorite also caused a substantial depletion of intracellular glutathione, whereas lipid oxidation was not very prominent. Sodium chlorite had a lower toxicity to ocular mammalian cells than benzalkonium chloride, which could be responsible for the different effects of long-term application in the eye. The fungal cells, which were most resistant to sodium chlorite, maintained higher percentage levels of intracellular glutathione during treatment than the mammalian cells. The results show that sodium chlorite can cause oxidative stress in cells, and suggest that cell damage is more likely to be due to interaction with thiol compounds than with cell membrane lipids. The study also provides important information about the differential resistance of ocular cells and microbes to various preservatives and oxidants. PMID- 12559985 TI - C-terminal Hsp-interacting protein slows androgen receptor synthesis and reduces its rate of degradation. AB - The androgen receptor (AR) is a member of the nuclear receptor superfamily that requires the action of molecular chaperones for folding and hormone binding. C terminal Hsp-interacting protein (Chip) is a cochaperone that interacts with Hsp70 and Hsp90 molecular chaperones via a tetratricopeptide domain and inhibits chaperone-dependent protein folding in vitro. Chip also stimulates protein degradation by acting as an E3 ubiquitin ligase via a modified ring finger domain called a U box. We analyzed whether Chip affected AR levels using a transient transfection strategy. Chip overexpression led to a large decrease in AR steady state levels and increased levels of AR ubiquitinylation. However, Chip effects were not fully reversed by proteasome inhibitors, suggesting that mechanisms alternative to or in addition to proteasome-mediated degradation were involved. This hypothesis was supported by the finding that Chip overexpression reduced the rate of AR degradation, consistent with an effect on AR folding, perhaps leading to aggregation. The possibility that Chip affected AR folding was further supported by the finding that the effects of exogenous Chip were reproduced by a mutant lacking the U box. These results are discussed in terms of the role played by molecular chaperones in AR biogenesis. PMID- 12559986 TI - Docosahexaenoic acid-containing phosphatidylethanolamine in the external layer of liposomes protects docosahexaenoic acid from 2,2'-azobis(2 aminopropane)dihydrochloride-mediated lipid peroxidation. AB - We have proposed that incorporation of docosahexaenoic acid (DHA) into phosphatidylethanolamine (PE) might enhance resistance to lipid peroxidation in vivo. In this study, we examined the relationship between the transbilayer distribution of PE and the oxidative stability of DHA in PE. Liposomes composed of a phospholipid mixture were used as models for biological membranes. To modulate the transbilayer distribution of PE obtained from the liver of rats fed DHA (PE-DHA), we used phosphatidylcholine (PC) with two types of acyl chain region: dipalmitoyl (PC16:0) or dioleoyl (PC18:1). The proportion of PE-DHA in the liposomal external layer was significantly higher in liposomes containing PC18:1 than in those containing PC16:0. This tendency was more pronounced in liposomes extruded using a polycarbonate filter with smaller pore sizes. Additionally, PE-DHA in the external layer of liposomes prepared using a filter with smaller pore sizes could protect DHA itself from 2,2(')-azobis(2 aminopropane)dihydrochloride-mediated lipid peroxidation. PMID- 12559987 TI - Methylglyoxal-induced glycation affects protein topography. AB - Methylglyoxal is a metabolic byproduct that is elevated in diabetic tissue. We examined the effects of methylglyoxal on cytosolic aspartate aminotransferase (cAAT), which is an enzyme previously shown to be modified by glyceraldehyde, acrolein, and ribose 5-phosphate. In the present study we observed that methylglyoxal caused real-time changes in tryptophan (intrinsic) fluorescence. Millimolar concentrations of methylglyoxal predominately decreased the fluorescence emission at 388 nm. While micromolar concentrations also decreased emission at 388 nm, low levels of methylglyoxal caused a prominent redshift in the wavelength of maximal emission. The changes in intrinsic fluorescence reflect definable changes in protein topography. These observations are consistent with a change in conformation that is more compact than that of native cAAT, suggesting that intramolecular cross-links (i.e., lysine-lysine) or hydrophobic pockets (i.e., carboxyethyl-lysines) were formed. Methylglyoxal also inhibited activity, and the inhibition correlated with the methylglyoxal-induced change in protein conformation. PMID- 12559988 TI - Structural modifications of the permeability transition pore complex in resealed mitochondria induced by matrix-entrapped disaccharides. AB - Mitochondrial resealing after the opening of the permeability transition (PT) pore was studied in saline- and sugar-based media by following the fluorescence anisotropy changes of mitochondria-bound hematoporphyrin (HP), a probe sensitive to conformational variations of the pore complex [Biochemistry 38 (1999) 9300]. The HP anisotropy changes correlated well with complete mitochondrial resealing in saline media and suggested that the pore complex regained the native structure after closure. Rebuilding of the pore complex structure was also achieved in monosaccharide-based media, thus ruling out a major influence of the swollen state of mitochondria on the reconstitution properties of the pore components. On the contrary, when sucrose or other disaccharides were used as osmotic support, restoration of the native mitochondrial structure, as monitored by HP anisotropy, was not achieved, though the proton barrier of the inner membrane and respiration functions were reestablished. Infrared spectroscopy experiments indicated the occurrence of strong perturbations of the mitochondrial membrane structure after disaccharide entrapment in the matrix space. These data suggest that mitochondria are able to reseal and regain functional activity after opening of the PT pore irrespective of the incubation medium but in sucrose (and other disaccharides) the pore complex adopts a conformation different from that existing before permeabilization. In general, our data indicate that the pore complex can exist in different conformations which are modulated by the nature of the interactions with the medium cosolvents. PMID- 12559989 TI - Transport of 5-formyltetrahydrofolate into primary cultured rat astrocytes. AB - Transport of 5-formyltetrahydrofolate (5-FTHF) into primary cultured rat astrocytes was studied. Uptake of 5-FTHF into astrocytes was linear in the first 60 min and is saturable with K(m)=3.3 microM and V(max)=27.5 pmol/mg protein/45 min in pH 7.4 medium. Uptake of 5-FTHF displayed the characteristics of countertransport. Uptake of 5-FTHF was inhibited by the structural analogs 5 methyltetrahydrofolate, methtrexate, and folic acid (K(i)=3.8, 2.7, and 18.4 microM, respectively). Uptake was significantly decreased by sodium azide but was increased by high concentration of sodium cyanide and low concentration of sodium arsenate. Uptake was also inhibited by p-chloromercuriphenylsulfonate and by the anions probenecid and 4,4(')-diisothiocyanostilbene-2,2(')-disulfonic acid. Acute exposure of the cells to ethanol (100mM) inhibited the uptake for 90 min of the experimental duration. It is concluded that astrocytes have a system for the uptake of 5-FTHF and folate analogs which is carrier mediated, this system is sensitive to energy inhibitors and alcohol exposure. PMID- 12559990 TI - Effect of gamma-synuclein overexpression on matrix metalloproteinases in retinoblastoma Y79 cells. AB - gamma-Synuclein is a small cytoplasmic protein implicated in neurodegenerative diseases and cancer. However, the mechanism of its involvement in diseases is not clear. We studied the role of gamma-synuclein in the regulation of matrix metalloproteinases in retinoblastoma cell culture. Matrix metalloproteinases play important roles in the remodeling of extracellular matrix implicated in tumor progression and in the neurodegenerative diseases. Western blot and zymography data demonstrated a moderate elevation of matrix metalloproteinases-2 and significant upregulation of matrix metalloproteinases-9 in stable cell lines overexpressing gamma-synuclein. No effect of gamma-synuclein overexpression on matrix metalloproteinases-1 level or activity was found. Chloramphenicol acetyltransferase assay demonstrated that overexpression of gamma-synuclein increases the efficiency of the matrix metalloproteinases-9 promoter. This increment of promoter activity may be mediated by the AP-1 binding site(s), since point mutations in one of these sites (Pr18 or Pr19) and elimination of the distal AP-1 site (Pr14) reduced the increment of promoter activity. PMID- 12559992 TI - A review of therapist characteristics and techniques positively impacting the therapeutic alliance. AB - The present review is a comprehensive examination of the therapist's personal attributes and in-session activities that positively influence the therapeutic alliance from a broad range of psychotherapy perspectives. Therapist's personal attributes such as being flexible, honest, respectful, trustworthy, confident, warm, interested, and open were found to contribute positively to the alliance. Therapist techniques such as exploration, reflection, noting past therapy success, accurate interpretation, facilitating the expression of affect, and attending to the patient's experience were also found to contribute positively to the alliance. This review reveals how these therapist personal qualities and techniques have a positive influence on the identification or repair of ruptures in the alliance. PMID- 12559991 TI - Molecular pathway for (-)-epigallocatechin-3-gallate-induced cell cycle arrest and apoptosis of human prostate carcinoma cells. AB - Epigallocatechin-3-gallate (EGCG), the major polyphenolic constituent present in green tea, is a promising chemopreventive agent. We recently showed that green tea polyphenols exert remarkable preventive effects against prostate cancer in a mouse model and many of these effects are mediated by the ability of polyphenols to induce apoptosis in cancer cells [Proc. Natl. Acad. Sci. USA 98 (2001) 10350]. Earlier, we showed that EGCG causes a G0/G1 phase cell cycle arrest and apoptosis of both androgen-sensitive LNCaP and androgen-insensitive DU145 human prostate carcinoma cells, irrespective of p53 status [Toxicol. Appl. Pharmacol. 164 (2000) 82]. Here, we provide molecular understanding of this effect. We tested a hypothesis that EGCG-mediated cell cycle dysregulation and apoptosis is mediated via modulation of cyclin kinase inhibitor (cki)-cyclin-cyclin-dependent kinase (cdk) machinery. As shown by immunoblot analysis, EGCG treatment of LNCaP and DU145 cells resulted in significant dose- and time-dependent (i) upregulation of the protein expression of WAF1/p21, KIP1/p27, INK4a/p16, and INK4c/p18, (ii) down modulation of the protein expression of cyclin D1, cyclin E, cdk2, cdk4, and cdk6, but not of cyclin D2, (iii) increase in the binding of cyclin D1 toward WAF1/p21 and KIP1/p27, and (iv) decrease in the binding of cyclin E toward cdk2. Taken together, our results suggest that EGCG causes an induction of G1 phase ckis, which inhibits the cyclin-cdk complexes operative in the G0/G1 phase of the cell cycle, thereby causing an arrest, which may be an irreversible process ultimately leading to apoptotic cell death. This is the first systematic study showing the involvement of each component of cdk inhibitor-cyclin-cdk machinery during cell cycle arrest and apoptosis of human prostate carcinoma cells by EGCG. PMID- 12559993 TI - The relevance of attachment theory to the philosophy, organization, and practice of adult mental health care. AB - This review is an inquiry into the relevance of attachment theory to the current philosophy, organization, and practice of adult mental health care, via an examination of the literature relating to attachment theory and, in particular, the literature relating to research into adult attachment. The review does not seek to critique attachment theory itself, but considers carefully the relevance of the theory to adulthood and to the field of adult mental health. In so doing, research into individual difficulties is examined, as is the provision and delivery of therapeutic services. In addition, the literature regarding the importance of mental health staff's own attachments and the influence of attachment theory on mental health service philosophy and organization are evaluated. Finally, potential areas for future research and development in this field are suggested. PMID- 12559994 TI - Psychiatric comorbidity in anorexia and bulimia nervosa: nature, prevalence, and causal relationships. AB - Eating disorders are complex, multifactorially determined phenomena. When individuals with eating disorders present for treatment with comorbid conditions, case conceptualization is further complicated and, as a result, it may be difficult to determine optimal psychological or pharmacological treatment. This article reviews the evidence of the association between eating disorders (anorexia nervosa [AN] and bulimia nervosa [BN]) and Axis I depression, obsessive compulsive disorder (OCD), substance abuse, and Axis II personality disorders, for the purposes of increasing awareness about the different options for case conceptualization. Although other diagnoses comorbid with eating disorders are of interest to clinicians (e.g., posttraumatic stress disorder [PTSD] and social phobia), their comprehensive review is currently premature due to a lack of empirical scrutiny. Finally, future directions for research, including suggestions for the use of particular assessment tools and more sophisticated research designs, are discussed. PMID- 12559995 TI - The dimensional view of personality disorders: a review of the taxometric evidence. AB - The dimensional view of personality disorders (PDs) represents these conditions as extreme variants of normal personality continua. This widely held view underpins efforts to characterize PDs in terms of established systems of personality description and to overhaul classification of PDs along dimensional lines. A review of 21 taxometric studies of PDs and related variables calls an unqualified version of this view into question. Analyses of the three PDs investigated to date strongly support taxonic (i.e., categorical or discontinuous) models. Implications for the conceptualization and classification of PDs are drawn. PMID- 12559996 TI - Neuropsychology of obsessive-compulsive disorder: a review and treatment implications. AB - The existing literature examining neuropsychological features of obsessive compulsive disorder (OCD) is reviewed. The accumulated research points to a deficit in organizational strategies in general, suggesting problems in executive functioning. The available research is inconsistent in identifying memory deficits in OCD. However, memory problems are most evident when tests are used that require an implicit organizational strategy. While the majority of the research reviewed involves adult samples, there is emerging evidence that these deficits are present in children as well. It is suggested here that the interaction between organizational strategy deficits and the effort to recall unstructured information contributes to doubting, an important feature of OCD. Implications of this body of research for behavior therapy are considered. PMID- 12560002 TI - Adolescents and their parents: a critical review of measures to assess their satisfaction with one another. AB - The importance of assessing the parent-adolescent relationship has been stressed in the literature. However, an integration of studies that have been conducted to assess satisfaction in the parent-adolescent relationship is warranted, including evaluation of measures to assess their satisfaction with one another. Therefore, the purpose of this paper is to provide a critical examination of the clinical utility and psychometric properties of such measures. Suggestions for future research are also provided. PMID- 12560003 TI - Evaluating meta-ethnography: a synthesis of qualitative research on lay experiences of diabetes and diabetes care. AB - Interest in how qualitative health research might be used more widely to inform health policy and medical practice is growing. Synthesising findings from individual qualitative studies may be one method but application of conventional systematic review methodology to qualitative research presents significant philosophical and practical challenges. The aim here was to examine the feasibility of synthesising qualitative research using qualitative methodology including a formative evaluation of criteria for assessing the research to be synthesised. Ten qualitative studies of adult patients' perspectives of diabetes were purposefully selected and questions proposed by the critical appraisal skills programme (CASP) adapted and used to assess papers prior to synthesis. Each study was reviewed independently by two experienced social scientists. The level of agreement between reviewers was determined. Three papers were excluded: one because it turned out not to be qualitative research, one because the quality of the empirical work was poor and one because the qualitative findings reported were also recorded in another paper already included. The synthesis, which had two distinct elements, was conducted using the meta-ethnographic method. Firstly, four papers containing typologies of patient responses to diabetes were synthesised. Secondly, six key concepts were identified from all seven papers as being important in enabling a person with diabetes to achieve a balance in their lives and to attain a sense of well-being and control. These included: time and experience, trust in self, a less subservient approach to care providers, strategic non-compliance with medication, effective support from care providers and an acknowledgement that diabetes is serious. None of the studies included in the synthesis referenced any of the early papers nor did they appear to have taken account of or built upon previous findings. This evaluation confirmed that meta-ethnography can lead to a synthesis and extension of qualitative research in a defined field of study. In addition, from it a practical method of qualitative research assessment evolved. This process is promising but requires further testing and evaluation before it could be recommended for more widespread adoption. PMID- 12560004 TI - Evaluation of midwifery care: results from a survey in rural Guatemala. AB - In an effort to reduce infant and maternal morbidity and mortality in developing countries, the World Health Organization has promoted the training of traditional birth attendants (midwives) and their incorporation into the formal health care system. In this paper, we examine several aspects of the integration of traditional and biomedical maternity care that are likely to reflect the quality of care received by Guatemalan women. Specifically, we examine the extent to which women combine traditional and biomedical pregnancy care, the frequency with which midwives refer women to biomedical providers, the content and quality of care offered by midwives, and the effects of midwife training programs on referral practices and quality of care. The analysis is based on data from the 1995 Guatemalan Survey of Family Health. The results offer a mixed assessment of the efficacy of midwife training programs. For example, although trained midwives are much more likely than other midwives to refer their clients to biomedical providers, most pregnant women do not see a biomedical provider, and the quality of midwife care, as defined and measured in this study, is similar between trained and untrained midwives. PMID- 12560005 TI - Rapid assessment procedures of malaria in low endemic countries: community perceptions in Jepara district, Indonesia. AB - Most studies on community perceptions toward malaria have been undertaken in high endemic countries, and studies from low-endemic countries have only recently been published. Similar information is also needed for hypoendemic countries such as Indonesia, to cope with the persistence of foci-endemic malaria in these regions. An applied qualitative method, Rapid Assessment Procedures, was employed during a 3-month intensive data collection period in Jepara district, Central Java province. Data were retrieved from 38 free-listings, 28 in-depth interviews, seven focus group discussions and unstructured observation. Qualitative thematic content analysis was applied. In this community, malaria (known as katisen or panas tis) was considered a common but minor illness. Insufficient understanding of malaria signs and symptoms in the subvillages likely leads to delay in illness recognition and treatment; not surprisingly self-treatment is common and the dosage most likely below the recommended dose. The health center was used but when it did not work, most people would shift back to traditional services due to cost considerations. Low understanding and acceptance of the causal link between the mosquito and malaria, likely leading to poor comprehension of preventive activities, as well as confusion of malaria with dengue fever, were identified. In conclusion, this study highlights a consistent gap between the common understanding and the biomedical description of malaria. If case management continues to be the main strategy in malaria control program, the emic perspective of the people must be well-integrated into the program. Likewise, interventions to improve home-treatment should also be developed. PMID- 12560006 TI - War, gender and culture: Mozambican women refugees. AB - Analyses of the psychological sequelae of war-related violence for women tend to rely on the concepts developed in research on male combatants. Post-traumatic stress disorder or varying combinations of its symptoms are identified as the principal outcomes of war-related events for women. By and large, the dominant literature does not examine possible outcomes which could be specified by gender. This paper refers to the war in Mozambique during the 1970s and 1980s as a typical illustration of how women are an integral part of the battlefield. It draws on research on African women and uses testimony of Mozambican women refugees who settled in South Africa to explore how gender is linked to psycho social outcomes of massive social conflict. The paper argues that a richer understanding of the psycho-social outcomes of war and the needs of survivors is promoted by investigating gender in specific historical situations and how this frames the responses people have to experiences of violence and social destruction. PMID- 12560007 TI - International NGOs and primary health care in Mozambique: the need for a new model of collaboration. AB - In keeping with the neo-liberal emphasis on privatization, international aid has been increasingly channeled through non-governmental organizations (NGOs) and their expatriate technical experts to support primary health care (PHC) in the developing world. Relationships between international aid workers and their local counterparts have thus become critical aspects of PHC and its effectiveness. However, these important social dynamics of PHC remain understudied by social scientists. Based on three years of participant-observation in Mozambique, this paper presents an ethnographic case study of these relationships in one central province. The Mozambique experience reveals that the deluge of NGOs and their expatriate workers over the last decade has fragmented the local health system, undermined local control of health programs, and contributed to growing local social inequality. Since national health system salaries plummeted over the same period as a result of structural adjustment, health workers became vulnerable to financial favors offered by NGOs seeking to promote their projects in turf struggles with other agencies. It is argued that new aid management strategies, while necessary, will not be sufficient to remedy the fragmentation of the health sector. A new model for collaboration between expatriate aid workers and their local counterparts in the developing world is urgently needed that centers on the building of long-term equitable professional relationships in a sustainable adequately funded public sector. The case study presented here illustrates how the NGO model undermines the establishment of these relationships that are so vital to successful development assistance. PMID- 12560008 TI - Subfecundity and anxiety in a nationally representative sample. AB - Research thus far on the psychological consequences of impaired fecundity in developed countries has relied heavily on clinic-based samples. This study uses a nationally representative sample of American women regardless of fecundity status or treatment status. I analyze reports of fecundity status and anxiety from a 1995 sample of almost 11,000 respondents. The results show consistent positive effects of subfecundity on the odds of fulfilling the diagnostic criteria for Generalized Anxiety Disorder and the incidence rate of symptoms among those who would be so diagnosed, even when controlling for potential confounding factors. Whether a subfecund respondent currently desires to have a child does not moderate the likelihood of being anxious, but does moderate the number of symptoms reported. The lack of a moderating effect of seeking treatment suggests that past research on clinic-based samples is generalizable to all subfecund women. PMID- 12560009 TI - Provider bias in the treatment of diarrhea among boys and girls attending public facilities in Minia, Egypt. AB - Girls' excess mortality in early childhood persists in all regions of the world and has been attributed to parental discrimination in the allocation of food and healthcare. Consequently, researchers have paid scant attention to the potential for unequal treatment of boys and girls by health providers. Data from a longitudinal study of child morbidity conducted between 1995 and 1997 in Minia, Egypt are used to compare the frequency with which diarrheal cases of boys and girls were treated with oral rehydration solution (ORS) among children ever presenting at a formal source of care. Multivariate analysis suggests that public providers have marginally higher adjusted odds of administering ORS to boys than to girls and significantly higher adjusted odds of giving and recommending ORS to the caretakers of boys than of girls. Findings expose a need for further research on the supply side determinants of girls' disadvantaged treatment in Upper Egypt and in other settings where son preference persists. PMID- 12560010 TI - The measurement of SES in health research: current practice and steps toward a new approach. AB - The resurgence of social epidemiology has yet to induce corresponding research into basic measurement issues. This paper aims to motivate investigators to refocus attention on the measurement of socioeconomic status (SES). With a primarily American focus, we document striking paucity of basic research in SES, review the history of SES measurement, highlight the central limitations of current measurement approaches, sketch a new theoretical perspective, present new pilot results, and outline areas for future research. We argue (1) that lack of conceptual clarity and the bypassing of standard psychometric techniques have retarded SES measurement. And (2) social epidemiologists should revisit the measurement of SES and consider whether a richer, psychometrically induced, approach would be more useful. Our pilot study suggests a great deal of uniformity between existing SES measures and that a new approach may be worthy of pursuit. PMID- 12560011 TI - Does active leisure protect cognition? Evidence from a national birth cohort. AB - Social, physical and intellectual activities are thought to facilitate cognitive performance and slow the rate of age associated cognitive decline, but little is known about this association in younger adulthood. We used multiple regression to test the association between two kinds of activity at 36 years-physical exercise and spare-time activity-and verbal memory at 43 and 53 years in 1919 males and females enrolled in the MRC National Survey of Health and Development (the British 1946 birth cohort). Both kinds of activities were significantly and positively associated with memory performance at 43 years, after controlling for sex, education, occupational social class, IQ at 15 years, and recurrent ill health and significant mental distress. Furthermore, physical exercise at 36 years (but not spare-time activity) was associated with a significantly slower rate of decline in memory from 43 to 53 years, after controlling for the same factors, with evidence that continuing physical exercise after 36 years was important for protection. We conclude that physical exercise and spare-time activity are significantly associated with benefit to memory in midlife, although these two kinds of voluntary activity may exert their effects on cognition via different paths. PMID- 12560012 TI - Burial at Srebrenica: linking place and trauma. AB - Five years after the massacre at Srebrenica in Bosnia-Herzegovina, survivors were faced with the decision: where did they want their loved ones buried? This report explores the reasons for their choice in qualitative interviews with 37 survivors of the massacre and 22 key informants performed over the summer 2000. Survivors wanted the loved ones buried at Potocari, a site just outside of Srebrenica, because it represented the site of ultimate horror, was connected to their sense of home, and underscored the various power relationships. The data points to the importance of place for health. Trauma, as it occurs in particular locations, breaks the sense of attachment to a particular place. Restoring the physical and social environment through burial and memorials mitigates the consequences of the trauma. The burial at Potocari provides a window into the mourning, politics, and recovery after mass violence. PMID- 12560013 TI - Eliciting preferences of the community for out of hours care provided by general practitioners: a stated preference discrete choice experiment. AB - Access to primary care services is a major issue as new models of delivering primary care continue develop in many countries. Major changes to out of hours care provided by general practitioners (GPs) were made in the UK in 1995. These were designed in response to low morale and job dissatisfaction of GPs, rather than in response to patients' preferences. The aim of this study is to elicit the preferences of patients and the community for different models of GP out of hours care. A questionnaire was sent to parents of children in Aberdeen and Glasgow in Scotland who had received a home visit or attended a primary care emergency centre, or were registered with a GP. The questionnaire used a discrete choice experiment that asked parents to imagine their child had respiratory symptoms. Parents were then asked to choose between a series of pairs of scenarios, with each scenario describing a different model of out of hours care. Each model varied by waiting time, who was seen, location, and whether the doctor listened. The response rate was 68% (3,893/5,718). The most important attribute was whether the doctor seemed to listen, suggesting that policies aimed at improving doctor patient communication will lead to the largest improvements in utility. The most preferred location of care was a hospital accident and emergency department. This suggests that new models of primary care emergency centres may not reduce the demand for accident and emergency visits from this group of patients in urban areas. Preferences also differed across sub-groups of patients. Those who had never used out of hours care before had stronger preferences for waiting time and the doctor listening, suggesting higher expectations of non-users. Further research is required into the demand for out of hours care as new models of care become established. PMID- 12560014 TI - Psychosocial burden of beta-thalassaemia major in Antalya, south Turkey. AB - beta-thalassaemia is a recessively inherited blood disorder characterised by chronic anaemia. It requires monthly blood transfusions and regular iron chelation. Thousands of affected children are born annually and the magnitude of the problem is most severe in developing countries. Ninety-nine children and 32 adults with thalassaemia major, and 112 parents of patients were interviewed in Antalya, south Turkey, using specifically designed questionnaires to evaluate psychosocial burden. The education of most of the thalassaemic children of school age (60%) was affected, mainly due to having to attend hospital for investigation and transfusions. A high level of parental anxiety (82%) was reported. Nearly half of the families (47%) had employment and financial problems as a result of thalassaemia, yet there was a low level of marital breakdown (1.8%). A substantial majority (93%) of the parental couples would have chosen to terminate an affected pregnancy if they had known that the foetus had thalassaemia major. The results reflect the need for a national policy for public education and screening of thalassaemia in Turkey in order to offer prenatal diagnosis for all families at risk of homozygous thalassaemia. PMID- 12560015 TI - In search of a healing place: Aboriginal women in Vancouver's Downtown Eastside. AB - Research on general health service delivery in urban areas of Canada shows that Aboriginal people face formidable barriers in accessing culturally appropriate and timely care. Over the past decade, Urban Aboriginal Health Centres (UAHCs) have emerged to address the unmet health concerns of Aboriginal people living in metropolitan areas of the country. The purpose of this research was to address the gap in social science literature on how the health care concerns of Aboriginal women are being met by UAHCs. The research aimed to give voice to Aboriginal women by asking them whether the appropriate professional services and educational programs they need to address their health care needs were being provided in the inner city. A case-study approach was used whereby three separate focus groups were conducted with Aboriginal women who were clients of the Vancouver Native Health Society (VNHS), its sister organization, Sheway, or residents of Vancouver's Downtown Eastside (DTES). In addition, twenty-five semi structured interviews were conducted with VNHS staff, health providers, government representatives, and community leaders in health care (total n=61). The findings indicate that despite efforts from various quarters to articulate the health and social concerns of the country's marginalized populations, such has not been the case for Aboriginal women living in one of Canada's most prosperous cities. Many Aboriginal women expressed a strong desire for a Healing Place, based on a model of care where their health concerns are addressed in an integrated manner, where they are respected and given the opportunity to shape and influence decision-making about services that impact their own healing. PMID- 12560016 TI - "Patient counselling" by pharmacists: four approaches to the delivery of counselling sequences and their interactional reception. AB - 'Patient counselling' by pharmacists is a diverse and ill-defined activity. It is also an activity which is achieving more prominence as part of the 'extended role' which is seen as the way forward for the profession. This paper uses data from a hospital paediatric outpatient clinic in the United Kingdom to examine the process of patient counselling from a conversation analytic standpoint, with a particular focus on the varying ways in which these sequences are set up and the ways in which patients or carers respond. Four types of interactional approach to negotiating entry into a broadly defined 'patient counselling' sequence are identified. These approaches are considered within the broader frameworks of delicacy, morality and competence which impact upon the giving and receiving of advice and information more generally, as well as in this setting, and in the light of the continued development of the 'extended role'. PMID- 12560017 TI - Ethnic differences in choices of health information by cancer patients using complementary and alternative medicine: an exploratory study with correspondence analysis. AB - This study examined patterns in the use of health information among Caucasian, Japanese, and non-Japanese Asian Pacific Islander cancer patients in Hawaii and explored the relation of ethnicity and educational level to choices of health information sources. Information from 140 cancer patients, most of whom were users of complementary and alternative medicine (CAM), was analyzed using correspondence analysis. Three clusters of health information pertinent to the three ethnic groups emerged from the data. The results of this study revealed that Caucasian patients preferred objective, scientific, and updated information obtained through medical journals or newsletters from research institutions, telephone information services, and the internet. Japanese patients relied on media and commercial sources including television, newspapers, books, magazines and CAM providers. Non-Japanese Asians and Pacific Islanders used information sources involving person-to-person communication with their physicians, social groups, and other cancer patients. A higher educational level was closely related to a cluster of health information stressing objective, scientific and updated information, while a lower educational level was associated with interpersonally communicated information. The three ethnicity-specific patterns of health information use remained relatively stable at different educational levels, implying that the effect of patients' ethnicity overrides their educational level in shaping their choices of health information. The results of this study indicate the importance of recognizing cancer patients' culturally developed world views when understanding their health information-seeking behavior. For medical practice, these findings indicate the need for healthcare providers to assist cancer patients to obtain accurate health information in a culturally sensitive way. PMID- 12560018 TI - Menstrual-related changes expected by premenarcheal girls living in rural and urban areas of Mexico. AB - Most women experience changes surrounding the start of menstruation. These changes are influenced by sociocultural context. Consequently, certain changes are more pronounced in some cultures than in others. Girls enter menarche with a clear set of paramenstrual expectations that may alter their menstrual cycle related experiences when they become postmenarcheal. This study explored expectations concerning the paramenstrual changes of 1,173 premenarcheal girls living in rural and urban areas of Mexico. In accordance with the findings of studies conducted in other countries, Mexican premenarcheal girls associate menstruation with a set of mostly negative expectations. A comparison of the results from urban and rural girls revealed that urban girls expected negative paramenstrual changes more, while rural ones expected positive changes more. These differences suggest that the cultures in which girls are brought up have an impact on their expectations. Urban girls are more exposed to media which present a picture of menses as a debilitating event, while rural girls link menses with health because it is associated with the ability to have children. PMID- 12560019 TI - The risk of smoking in relation to engagement with a school-based smoking intervention. AB - Health promotion interventions cannot work if people do not engage with them. The aim of this study was to examine whether disengagement from an adolescent smoking prevention and cessation intervention was an independent risk factor for regular smoking 1 and 2 years later. The data were taken from a cluster randomised controlled trial, in the West Midlands, UK, based on the transtheoretical or stages of change model. In this trial, 8,352 13-14-year old school pupils enrolled, and the data in this report were based on the 7,413 and 6,782 pupils present at 1 and 2 years follow-ups, respectively. The intervention group undertook three sessions using an interactive computer programme. At the end of the programme, pupils recorded their responses to it. Pupils were classed as engaged if they thought the intervention was both useful and interesting; all others were classed as disengaged. Random effects logistic regression related the number of times engaged to regular smoking at 1 and 2 years follow-up, adjusted for school absences and 11 potential confounders. The majority of pupils were engaged by the intervention. For participants using the intervention three times but not engaging once, the odds ratios (95% confidence intervals) for smoking at 1 and 2 years relative to the controls were 1.83 (1.41-2.39) and 1.70 (1.38 2.11). For those engaging three times, they were 0.79 (0.60-1.03) and 0.96 (0.75 1.21). There was no interaction with baseline intention to smoke, classified by stage of change, but there was a borderline significant interaction with baseline smoking status, with disengagement acting as a stronger risk factor among baseline never-smokers. We conclude that disengagement from interventions is a risk factor for smoking independently of experimentation with cigarettes. The best explanation is that disengagement from school, an established risk factor for smoking, generalises to disengagement from didactic school-based health promotion programmes. PMID- 12560020 TI - Gender differences in excessive daytime sleepiness among Japanese workers. AB - Excessive daytime sleepiness (EDS) is serious concern in the workplace with respect to errors, accidents, absenteeism, reduced productivity and impaired personal or professional life. Previous community studies found a female preponderance of EDS, however, there is little research on EDS and gender in occupational settings. We examined the gender differences in prevalence and risk factors of EDS among employees working at a telecommunications company in the Tokyo metropolitan area. Our outcome measure of EDS was the Epworth Sleepiness Scale (ESS). A self-administered questionnaire on health and sleep including ESS was distributed to 5,571 workers between December 1999 and January 2000, and 5,072 responses were returned (91.0%). A total of 4,722 full-time, non-manual and non-shift employees aged 20-59 were used for analysis (3,909 men and 813 women). Chi-squared tests and multiple logistic regression analyses were applied for examining the gender differences in the prevalence and risk factors of EDS. The prevalence rates of EDS were 13.3% for women and 7.2% for men (P<0.001). We identified that deprived nocturnal sleep, an irregular sleep-wake schedule and depression were the risk factors of EDS for both genders, and being married worked as a protective factor against EDS for men alone. It is obvious that a ban on overtime work and a provision of mental health hygiene are the general strategies for reducing EDS at worksites. In the case of women, we suggest the formation of effective strategies for improving women's status at home and in the workplace must also be a solution for the prevention of EDS (e.g. promoting gender equality in the division of labor at home and strengthening family care policies for working women). PMID- 12560021 TI - Commentary on: A psychometric analysis of the measurement level of the rating scale, time trade-off, and standard gamble, by Cook et al. PMID- 12560022 TI - A lupane triterpene from frankincense (Boswellia sp., Burseraceae). AB - A new lupane-type triterpene, 3alpha-hydroxy-lup-20(29)-en-24-oic acid, was isolated from the methanolic extract of "Erytrean-type" resin of commercial frankincense together with the known 3alpha-hydroxy-olean-12-en-24-oic acid (alpha-boswellic acid) and 3alpha-hydroxy-urs-12-en-24-oic acid (beta-boswellic acid). Their structures were characterized on the basis of chemical and spectral evidence including two dimensional NMR experiments and mass spectrometric techniques. PMID- 12560023 TI - Norditerpenoid alkaloids from Delphinium species. AB - From the aerial parts of four Delphinium species 11 known and 3 new norditerpenoid alkaloids have been isolated: from D. dissectum Huth: delavaine A/B, deoxylycoctonine, methyllycaconitine; new: 10-hydroxymethyllycaconitine; from D. excelsum Reichenb.: delcaroline, delectinine, delterine, methyllycaconitine; new: 10-hydroxymethyllycaconitine, 18-O-methyldelterine and 10-hydroxynudicaulidine; from D. grandiflorum L.: delcosine, deltatsine, grandiflorine, methyllycaconitine; from D. triste Fisch.: delcosine, macrocentridine, 14-dehydrodelcosine. The structures of the new alkaloids were established on the basis of MS, 1H, 13C, DEPT, homonuclear COSY, HMQC and HMBC NMR spectroscopic techniques. PMID- 12560024 TI - Neo-clerodane diterpenoids from Croton schiedeanus. AB - Two new neo-clerodane type furano diterpenoids were isolated from the aerial part of Croton schiedeanus, besides the clerodane diterpenes cis- and trans dehydrocrotonin, previously isolated from other species of Croton. Structural elucidation was achieved on basis of extensive NMR experiments, including X-ray diffraction analysis and molecular mechanics calculations. The previously known flavonoids ayanin and quercetin-3,7-dimethyl ether were also obtained from the extract of this plant. PMID- 12560025 TI - Triterpenoid saponins from Meryta lanceolata. AB - Four new oleanane-type saponins and a known one were isolated from the leaves and stems of Meryta lanceolata. The new saponins were characterised by spectroscopic means and chemical hydrolysis as 3-O-[beta-D-glucopyranosyl-(1-->3)-beta-D glucopyranosyl-(1-->3)-[beta-D-glucopyranosyl-(1-->2)]-alpha-L arabinopyranosyl]oleanolic acid 28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester, 3-O-[beta-D- glucopyranosyl (1-->3)-beta-D-glucopyranosyl-(1-->3)-[beta-D-glucopyranosyl-(1-->2)]-alpha-L arabinopyranosyl]oleanolic acid 28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-6-O acetyl glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester, 3-O-[beta-D glucopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->3)-beta-D-glucopyranosyl-(1- >3)-alpha-L-arabinopyranosyl]oleanolic acid 28-O-[alpha-L-rhamnopyranosyl-(1-->4) beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester and 3-O-[beta-D glucopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->3)-beta-D-glucopyranosyl-(1- >3)-alpha-L-arabinopyranosyl]echinocystic acid 28-O-[alpha-L-rhamnopyranosyl-(1- >4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester. The NMR assignments were made by means of HOHAHA, 1H-1H COSY, HMQC, HMBC and NOE difference studies. PMID- 12560026 TI - Bidesmosidic saponins from the fruits of Diploclisia glaucescens. AB - Chemical investigation of methanol extract of the fruits of Diploclisia glaucescens (Menispermaceae) furnished two new bidesmosidic saponins 3-O-[beta-D glucopyranosyl-(1-->3)-beta-D-glucopyranosyl]phytolaccagenic acid 28-O-beta-D glucopyranosyl ester, 3-O-[alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl (1-->2)-beta-D-glucopyranosyl]phytolaccagenic acid 28-O-beta-D-glucopyranosyl ester, together with known 3-O-beta-D-glucopyranosylphytolaccagenic acid 28-O beta-D-glucopyranosyl ester and 3-O-[alpha-L-rhamnopyranosyl-(1-->2)-beta-D glucopyranosyl-(1-->2)-beta-D-glucopyranosyl]serjanic acid 28-O-beta-D glucopyranosyl ester. The last saponin is reported for the first time from the family Menispermaceae. PMID- 12560027 TI - Ent-pimarane type diterpenes from Gnaphalium gaudichaudianum. AB - Fractionation of the methanol extract of Gnaphalium gaudichaudianum DC afforded one new and six known ent-pimarane diterpenes together with velutin, squalene and stigmasterol. The structure of the new compound was established on the basis of extensive 1D and 2D NMR spectroscopic data interpretation. Two of the isolated compounds exhibited moderate toxicity in the Artemia salina toxicity test. PMID- 12560028 TI - Pentacyclic triterpenoids from Embelia schimperi. AB - Five oleanane-type pentacyclic triterpenoids were isolated by chromatographic separation of a chloroform extract of the stem bark of Embelia schimperi. Three of these compounds have a methyleneoxy bridge. Two compounds, embelinone and schimperinone, are reported here for the first time from a natural source (they have been synthesized previously during chemical transformations). Their structures were determined by spectroscopic techniques, among which 2-D NMR was useful for complete characterization. Three of the triterpenoids exhibited mild antibacterial properties against the gram-positive bacterial strain Rhodococcus sp. PMID- 12560029 TI - Quassinoids from Ailanthus excelsa. AB - Three quassinoids, 1, 2 and 3, 4-dihydro excelsin 3 were isolated from the stem bark of Ailanthus excelsa, along with five known quassinoids excelsin, glaucarubine, ailanthinone, glaucarubinone and glaucarubolone. The glaucarubolone has been isolated for the first time from this plant. The structural elucidation is based on the analysis of spectroscopic data. PMID- 12560030 TI - Seco diterpenoids from Excoecaria agallocha L. AB - Chemical examination of the ethyl acetate solubles of the CH(3)OH:CH(2)Cl(2) (1:1) extract of the roots of Excoecaria agallocha L. collected from Godavary estuary resulted in the isolation of three more new diterpenoids, agallochins M-O (1-3). The structures of the new diterpenoids were elucidated by a study of their physical and spectral data as methyl ent-13-epi-8,13-epoxy-4,6alpha-dihydroxy-3,4 secolabd-14-en-3-oate (1), methyl ent-13-epi-8,13-epoxy-2,3-secolabd-14-en-2,11 olid-3-oate (2), and methyl ent-17-hydroxy-3,4-secokaura-4(19),15-dien-3-oate (3). PMID- 12560031 TI - Triterpenoids from the orchids Agrostophyllum brevipes and Agrostophyllum callosum. AB - Agrostophyllinol and agrostophyllinone, two new triterpenoids, were isolated from the orchid Agrostophyllum brevipes. Agrostophyllinone was also isolated from another orchid Agrostophyllum callosum. The structures of agrostophyllinol and agrostophyllinone were established as 24-methylene-lanosta-9(11)-en-3beta-ol (5a) and 24-methylene-lanosta-9(11)-en-3-one (5c), respectively, from spectral and chemical evidence. The above triterpenoids are of considerable biogenetic importance. PMID- 12560032 TI - Cardiobutanolide, a styryllactone from Goniothalamus cardiopetalus. AB - A styryllactone namely cardiobutanolide was isolated from the stem bark of Goniothalamus cardiopetalus together with four known styryllactones goniothalamin, goniodiol, goniofufurone, goniofupyrone and known acetogenins squamocin and an epimeric mixture of goniodonin and 34-epi-goniodonin. The structure of the new compound was elucidated on the basis of 1D and 2D NMR experiments and mass spectroscopic techniques. PMID- 12560033 TI - Stilbene derivatives from Gnetum gnemon Linn. AB - Four stilbene derivatives, gnemonols K and L (resveratrol trimers), M (isorhapontigenin dimer), and gnemonoside K (glucoside of resveratrol trimer) together with eleven known stilbenoids and a lignan were isolated from the acetone, methanol and 70% methanol soluble parts of the root of Gnetum gnemon (Gnetaceae). The structures of the isolates were determined by spectral analysis. The antioxidant activity of the stilbenoids on lipid peroxide inhibition and super oxide scavenging activity were also investigated. PMID- 12560034 TI - Prenylated xanthone glucosides from Ural's lichen Umbilicaria proboscidea. AB - Two new compounds isolated from an extract of a Central Asian lichen [Umbilicaria proboscidea (L.) Schrader=Syn.: Gyrophora proboscidea (L.) Ach.] are glucosides with mono- and di-prenylated xanthones as the aglycones and a saccharide moiety from two glucoses linked at C-7. The structures were elucidated on the basis of extensive spectroscopic analysis (1D and 2D NMR, MS, IR and UV) and by hydrolysis. PMID- 12560035 TI - Pharmacognostic studies on ginger and related drugs--part 1: five sulfonated compounds from Zingiberis rhizome (Shokyo). AB - Five sulfonated compounds, namely 4-gingesulfonic acid and shogasulfonic acids A, B, C and D, were isolated together with seven known compounds including 6 gingesulfonic acid from Zingiberis rhizome (Japanese name: Shokyo) made out of ginger. Their structures were characterized by means of spectroscopic analysis. PMID- 12560036 TI - Cytotoxic naphthoquinones and plumbagic acid glucosides from Plumbago zeylanica. AB - Two plumbagic acid glucosides, 3'-O-beta-glucopyranosyl plumbagic acid and 3'-O beta-glucopyranosyl plumbagic acid methylester along with five naphthoquinones (plumbagin, chitranone, maritinone, elliptinone and isoshinanolone), and five coumarins (seselin, 5-methoxyseselin, suberosin, xanthyletin and xanthoxyletin) were isolated from the roots of Plumbago zeylanica. All coumarins were not previously found in this plant. Cytotoxicity of these compounds to various tumor cells lines was evaluated, and plumbagin significantly suppressed growth of Raji, Calu-1, HeLa, and Wish tumor cell lines. PMID- 12560037 TI - Indolomonoterpenic alkaloids from Strychnos icaja roots. AB - In the course of our search for new antiplasmodial alkaloids from Strychnos icaja, we have isolated five alkaloids: three monomers, protostrychnine and genostrychnine, previously described in Strychnos nux-vomica, pseudostrychnine, already found in the leaves of the plant, a new bisindolic alkaloid, named strychnogucine C, and the first naturally occurring trimeric indolomonoterpenic alkaloid: strychnohexamine. This latter trimeric alkaloid presented an antiplasmodial activity against the FCA Plasmodium falciparum line near 1 microM. PMID- 12560038 TI - Ancistrolikokine D, a 5,8'-coupled naphthylisoquinoline alkaloid, and related natural products from Ancistrocladus likoko. AB - A new naphthylisoquinoline alkaloid, ancistrolikokine D, and the likewise 5,8' coupled alkaloid ancistroealaine A, as well as two further, biosynthetically related, but nitrogen-free natural products, ancistronaphthoic acid B and cis isoshinanolone, have been isolated from Ancistrocladus likoko J. LEACUTE;ONARD (Ancistrocladaceae). The 5,8'-coupling of the new alkaloids and of the alkaloids isolated earlier hints at a close phylogenetic relationship of A. likoko to other Central African Ancistrocladus species. The compounds show moderate activities against Leishmania donovani, Trypanosoma cruzi, and Trypanosoma brucei rhodesiense. PMID- 12560039 TI - Antifungal constituents of the stem bark of Bridelia retusa. AB - Antifungal activity guided fractionation of solvent extracts of the stem bark of Bridelia retusa of the family Euphorbiaceae against Cladosporium cladosporioides, furnished new bisabolane sesquiterpenes, (E)-4-(1,5-dimethyl-3-oxo-1 hexenyl)benzoic acid, (E)-4-(1,5-dimethyl-3-oxo-1,4-hexadienyl) benzoic acid, (R) 4-(1,5-dimethyl-3-oxo-4-hexenyl)benzoic acid and (-)-isochaminic acid, together with the known (R)-4-(1,5-dimethyl-3-oxohexyl)benzoic acid (ar-todomatuic acid), 5-allyl-1,2,3-trimethoxybenzene (elemicin), (+)-sesamin and 4-isopropylbenzoic acid (cumic acid). All these compounds showed fungicidal activity on TLC bioautography method at very low concentrations except elemicin. PMID- 12560040 TI - Polyacetylene glycosides from Pratia nummularia cultures. AB - Two polyacetylene glycosides, lobetyol 9-O-glc(6)-(1)rha (pratialin-A) and lobetyol 9-O-glc(6)-(1)glc(6)-(1)glc (pratialin-B), were isolated from Pratia nummularia (Campanulaceae) callus and hairy root cultures and their chemical structures were determined by analysis of spectroscopic data. From the methanol extract of the hairy root cultures, together with the known polyacetylene constituents lobetyol, lobetyolin, and lobetyolinin, tryptophan was also isolated. This report is the first example of the isolation and structure elucidation of rutinoside (pratialin-A) and triglucoside (pratialin-B) derivatives of polyacetylene constituents. PMID- 12560041 TI - Scaphopetalone and scaphopetalumate, a lignan and a triterpene ester from Scaphopetalum thonneri. AB - From the methanol extract of the stem bark of Scaphopetalum thonneri, two new compounds, including one lignan, named scaphopetalone, one new ester of ferulic acid, named scaphopetalumate were isolated together with three known compounds including: two coumarins (scopoletin and scopolin), and one pentacyclic triterpene (oleanolic acid). The structure of the new compounds were elucidated by means of spectroscopic analyses. PMID- 12560042 TI - Abstracts of the 6th International Congress of Comparative Physiology and Biochemistry. Mt. Buller, Australia, 2-7 february 2002. PMID- 12560043 TI - A selective HPLC method for determination of lercanidipine in tablets. AB - An HPLC reversed phase method using both UV (356 nm) and electrochemical (1000 mV) detection was developed in order to determine lercanidipine in commercial tablets. Repeatability and reproducibility were adequate. For quantification we have used the calibration plot method for lercanidipine concentration ranging between 1 x 10(-5) and 1 x 10(-4) M. Also, the proposed method is sufficiently selective to distinguish the parent drug and the degradation products after hydrolysis, photolysis or chemical oxidation. Furthermore, the typical excipients included in the drug formulation (talc, lactose, cornstarch, microcrystalline cellulose, carboxymethylcellulose and magnesium stearate) do not interfere with the selectivity of the method. Finally, the proposed chromatographic method was successfully applied to the quantitative determination of lercanidipine in commercial tablets. PMID- 12560044 TI - Determination of potassium ions in pharmaceutical samples by FIA using a potentiometric electrode based on ionophore nonactin occluded in EVA membrane. AB - A simple and rapid method was developed for the K(+) ions determination employing a flow injection system using a flow-through electrode based on the naturally occurring antibiotic ionophore nonactin occluded in a polymeric membrane. The nonactin ionophore was trapped in poly(ethylene-co-vinyl acetate) (EVA) matrix (40% w/w in vinyl acetate) and dispersed on the surface of a graphite-epoxy tubular electrode. The plasticizer-free all-solid-state potassium-selective electrode showed a linear response for K(+) concentrations between 5.0 x 10(-5) and 5.0x10(-2) M (r=0.9995) with a near-Nernstian slope of 51.5 mV per decade, when Tris-HCl buffer (pH 7.0;0,1 M) was employed as a carrier. The potentiometric FIA system allows an analytical frequency of 120 samples per hour with a precision of 3.6%. The relative standard deviations (R.S.D.) for K(+) determination in pharmaceuticals samples, without any previous treatment, were lower than 4.0%, comparable to those obtained by flame photometry. Ammonium is the main analytical interference and the electrode response time was 5 s at 25 degrees C. The useful lifetime of the tubular sensor is longer than 3 months in continuous use. PMID- 12560045 TI - The determination of a potential impurity in Thalidomide drug substance and product by HPLC with indirect UV detection. AB - Thalidomide molecule, a synthetic derivative of glutamine, can undergo hydrolysis at physiologic pH to form glutamine. Additionally, L-glutamine is one of the starting materials in the synthesis of Thalidomide drug substance. The current USP method for testing glutamine is thin-layer chromatography (TLC) with ninhydrin spray visualization. A more quantitative and automated high performance liquid chromatography (HPLC) method utilizing indirect ultraviolet (UV) detection was developed and validated for the determination of the non-UV absorbing glutamine in Thalidomide drug substance and product. The HPLC mobile phases consisted of phosphoric acid, 2-naphthalenesulfonate sodium and methanol. 2 Naphthalenesulfonate was used as a UV detection probe for glutamine. A segmented isocratic elution program was used to elute glutamine and Thalidomide, respectively. The method was found to be specific for glutamine. The linearity was 0.05-1.25% glutamine with respect to a nominal concentration of 8 mg ml(-1) Thalidomide sample. The limits of detection and quantitation were found to be 0.03 and 0.05% glutamine, respectively. The injection precision was 2.7% for area responses and 0.2% for the retention times. The recovery of glutamine at three concentration levels was found to be 100.8+/-2.8% from placebo and 99.2+/-5.8% from spiked Thalidomide drug substances. This newly developed HPLC method was used to determine glutamine in Thalidomide drug substances and products. The results from HPLC were in agreement with those from TLC. Therefore, the method developed is a suitable alternative to the current USP TLC procedure. Additionally, the method offers the advantage of being quantitative and automated. PMID- 12560046 TI - LC-MS and NMR determination of a dichloromethane artifact adduct, cyproheptadine chloromethochloride. AB - The British Pharmacopoeia (BP) monograph for cyproheptadine HCl tablets requires a 'Related substances' thin-layer chromatography (TLC) test. This test revealed an extraneous spot with an R(f) of 0.1 in certain cyproheptadine HCl tablets that were under ambient retention conditions as well as those on stability programs. An investigation utilizing LC-MS, direct infusion MS, NMR, and organic synthesis has identified that the spot results from the N-oxide of cyproheptadine (a genuine degradate) and a co-eluting cyproheptadine-dichloromethane adduct, an artifact formed during the sample extraction step in which dichloromethane is used in the extracting solvent. PMID- 12560047 TI - Simultaneous determination of naphazoline, diphenhydramine and phenylephrine in nasal solutions by capillary electrophoresis. AB - A capillary zone electrophoresis (CZE) method has been developed to separate and quantitate naphazoline (NAPH), dyphenhydramine (DIP) and phenylephrine (PHE) in nasal solutions. Samples were diluted 1:25 in ultrapure water and injected at the anodic end. A central composite design has been used to optimise the experimental conditions for a complete and fast separation of the active ingredients studied. Critical parameters such as voltage, pH and buffer concentration have been studied to evaluate how they affect responses such as resolution and migration times. Separation was performed on a silica capillary with 75 microm I.D. and 70 cm total length at an applied voltage of 17.7 kV with a phosphate run buffer of pH 3.72 and 0.063 mol l(-1). Calibration curves were prepared for NAPH, DIP and PHE. For each analyte, the correlation coefficients were >0.999 (n=15). The RSD% of six replicate injections for each analyte were reasonably good. The method was applied to the quantitation of the three components in a commercial dosage form. The proposed method has the advantage of needing a very simple sample pretreatment and being faster than a typical HPLC chromatographic method. PMID- 12560048 TI - Determination of 3'-C-ethynylcytidine in human plasma and urine by liquid chromatographic-electrospray ionization tandem mass spectrometry. AB - A liquid chromatographic-electrospray ionization tandem mass spectrometric (LC ESI/MS/MS) method was developed for the quantitative analysis of a novel anticancer drug, 3'-C-ethynylcytidine (I) in human plasma and urine. I and its stable isotope-labeled internal standard (II) were extracted from human plasma and urine samples using a polymer-based cation-exchange cartridge, and LC ESI/MS/MS analysis was performed by monitoring the positive fragment ions of I and II. The linear ranges are 1-500 ng/ml in plasma and 10-5000 ng/ml in urine. The limits of quantitation for I were 1 ng/ml in plasma and 10 ng/ml in urine. The relative errors (RE) for I ranged from -8.4 to 3.0% in plasma and from 0.8 to 4.4% in urine. The relative standard deviations (RSD) for I ranged from 1.2 to 8.9% in plasma and from 0.7 to 2.8% in urine. This validated analytical method is demonstrated to be useful for the analysis of I in human plasma and urine in clinical studies. PMID- 12560049 TI - LC method for the determination of oxcarbazepine in pharmaceutical preparations. AB - A simple and accurate HPLC method for determination of oxcarbazepine (OXC) in a new tablet formulation is described. Chromatographic separation was achieved on a Diamonsil C18 column using a mobile phase consisting of acetonitrile, potassium phosphate monobasic buffer (pH 6.8) and water (36:8:56, v/v) at a flow rate of 1.0 ml/min. Absorbance was monitored at 255 nm where OXC has maximum absorption. The linear range of detection for OXC was from 9.96 to 99.6 microg/ml. The proposed method was validated for selectivity, precision, accuracy and limits of detection and quantitation, etc. PMID- 12560050 TI - Utilizing capillary gas chromatography mass spectrometry to determine 4 benzotrifluoride t-butyl ether as a reaction by-product in fluoxetine synthesized using potassium t-butoxide as base. AB - Fluoxetine hydrochloride has been prepared using two similar synthetic routes, both of which rely upon an ether formation reaction mediated by a base. The base used can affect the impurity profile of this reaction. It was proposed that the synthesis of fluoxetine carried out using potassium t-butoxide as base and 4 chlorobenzotrifluoride (or 4-fluorobenzotrifluoride) in the ether formation step may result in the formation of 4-benzotrifluoride t-butyl ether as a reaction by product. To test this hypothesis, capillary gas chromatography-mass spectrometry (GC/MS) was utilized to evaluate samples of free base fluoxetine synthesized using sodium hydride (NaH) or potassium t-butoxide as the base. Assay conditions using selected ion monitoring (SIM) were developed, which allowed detection of trace levels (parts per million, ppm) of 4-benzotrifluoride t-butyl ether in fluoxetine free base sample matrix. Response linearity, precision, and standard spike recovery were examined during development, and were found to be suitable. Comparisons of fluoxetine samples generated from both NaH and potassium t butoxide processes were performed using the GC/MS assay. PMID- 12560051 TI - Determination of hydroxyanthraquinoids in Rhubarb by cyclodextrin-modified micellar electrokinetic chromatography using a mixed micellar system of sodium dodecyl sulfate and sodium cholate. AB - A cyclodextrin-modified micellar electrokinetic chromatographic (CD-MEKC) method was established to determine five hydroxyanthraquinoids in Rhubarb. The five components were successfully separated by using the mixed micellar system consisting of 20 mmol/l sodium dodecyl sulfate (SDS) and 20 mmol/l sodium cholate (SC) with 10 mmol/l beta-cyclodextrin in phosphate buffer (pH 10.4). The separation was optimized by adjusting buffer pH, concentrations of beta cyclodextrin and SC and applied voltage. The proposed method was validated and applied to the determination of two commercial Rhubarb samples. The results obtained were satisfactory. PMID- 12560052 TI - Biosensor analysis of the molecular interactions of pentosan polysulfate and of sulfated glycosaminoglycans with immobilized elastase, hyaluronidase and lysozyme using surface plasmon resonance (SPR) technology. AB - Pentosan polysulfate (NaPPS) and chondroitin sulfates (ChSs) have recently been shown to exhibit both symptom and disease modifying activities in osteoarthritis (OA), but their respective mechanisms of action are still the subject of conjecture. Excessive catabolism of joint articular cartilage is considered to be responsible for the initiation and progression of OA but the abilities of these drugs to mitigate this process has received only limited attention. Human neutrophil elastase (HNE) is a proteinase, which can degrade the collagens and proteoglycans (PGs) of the cartilage directly or indirectly by activating latent matrix metalloproteinases. Hyaluronidase (HAase) is an endoglycosidase, which degrades glycosaminoglycans including hyaluronan, which provides the aggregating component of the PG aggrecan complex. In the present study the molecular interactions between the NaPPS, ChSs and some other sulfated polysaccharides with immobilized HNE, HAase or lysozyme (a cationic protein implicated in PG metabolism) were studied using a SPR biosensor device-BIAcore2000. The above three enzymes were covalently immobilized to a biosensor chip CM5 separately using amine coupling. The binding affinity of each sulfated polysaccharide and the kinetics of NaPPS over the concentration range of 0.3-5.0 microg/ml were determined. The inhibition of HNE by the sulfated polysaccharides as determined using the synthetic substrate succinyl-Ala-Ala-Val-nitroanilide (SAAVNA) in a functional assay was compared with their respective binding affinities for this proteinase using the BIAcore system. The results obtained with the two independent techniques showed good correlation and indicated that the degree and ring positions of oligosaccharide sulfation were major determinants of enzyme inhibitory activity. The observed difference in order of binding affinities of the drugs to the immobilized HNE, HAase and lysozyme suggests a conformational relationship, in addition to the charge interactions between the sulfate esters of the polysaccharides and the cationic amino acids of the enzymes. Significantly, the SPR biosensor technology demonstrated that small differences among sulfated polysaccharides, even subtle variations among different NaPPS batches, could be readily detected. The SPR technology therefore offers not only a sensitive and reproducible method for ranking noncompetitive enzyme inhibitors for drug discovery but a rapid and quantitative bioassay for monitoring batch consistency of manufacture. PMID- 12560053 TI - In vitro metabolism of the new anxiolytic agent, RWJ-52763 in human hepatic S9 fraction-API-MS/MS identification of metabolites. AB - The in vitro metabolism of the anxiolytic agent, RWJ-52763 was studied after incubation with human hepatic S9 fraction in the presence of an NADPH-generating system. Unchanged RWJ-52763 (64% of the sample) plus six metabolites (M1-M6) were profiled, quantified, and tentatively identified on the basis of API-MS/MS data. The metabolic pathways for RWJ-52763 are proposed, and the two metabolic pathways are: (1) N/O-dealkylation, and (2) phenylhydroxylation. Pathway 1 formed a major N-dealkylated metabolite, N-desethoxy-RWJ-52763 (M1, 22% of the sample) and 2 minor N/O-dealkylated metabolites, O-desmethyl-RWJ-52763 (M2; 2%) and N,N didesethoxymethyl-RWJ-52763 (M3; 3%). Pathway 2 produced two hydroxyphenyl metabolites, hydroxydifluorophenyl-RWJ-52763 (M4; 4%) and hydroxyphenyl-pyrido RWJ-52763 (M5; 3%) in small amounts, and in conjunction with step 1 formed a minor N-desethoxymethyl-M4 (M6; 1%). RWJ-52763 is substantially metabolized by this human hepatic S9. PMID- 12560054 TI - A validated SPE-LC-MS/MS assay for Eplerenone and its hydrolyzed metabolite in human urine. AB - An automated LC-MS/MS assay was validated to quantitate the first selective aldosterone blocker Eplerenone (I) and its hydrolyzed metabolite (II) in human urine. After the addition of the stable isotope labeled internal standards, human urine samples were extracted on a C(18) solid phase extraction (SPE) cartridge using a Zymark RapidTrace automation system. The extraction eluates were diluted with 20 mM ammonium acetate aqueous solution and directly injected onto the LC MS/MS system. The chromatographic separation was performed on a reverse phase Zorbax XDB-C(8) HPLC column (2.1 x 50 mm, 5 microm) with a mobile phase of acetonitrile:water (40:60, v/v) containing 10 mM ammonium acetate (pH 7.4). I and II were ionized using positive and negative ionization mass spectrometry, respectively, to achieve the best sensitivity. The ionization polarity was switched during the run at approximately 2.5 min after the injection. Multiple reaction monitoring (MRM) with a tandem mass spectrometer was used to detect the analytes. The precursor to product ion transitions of m/z 415-->163 and m/z 431- >337 were used to measure I and II, respectively. The assay exhibited a linear dynamic range of 50-10000 ng/ml of urine for both of I and II. The lower limit of quantitation (LLOQ) was 50 ng/ml for I and II. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. Sample analysis time for each injection was 5 min; a throughput of 100 human urine standards and samples per run was achieved. PMID- 12560055 TI - On-line preconcentration and determination of chromium in parenteral solutions by flow injection-flame atomic absorption spectrometry. AB - An on-line chromium preconcentration and determination system implemented with flame atomic absorption spectrometry (FAAS) associated to flow injection (FI) was studied. For the retention of chromium, 4-(2-Thiazolylazo)-resorcinol (TAR) and Amberlite XAD-16 were used, at pH 5.0. The Cr-TAR complex was removed from the micro-column with ethanol. An enrichment factor of 50 was obtained for the preconcentration of 50 ml of sample solution. The detection limit value for the preconcentration of 50 ml of aqueous solution of Cr was 20 ng l(-1). The precision for ten replicate determinations at the 5 microg l(-1) Cr levels was 2.9% relative standard deviation (RSD), calculated from the peak heights obtained. The calibration graph using the preconcentration system for chromium was linear with a correlation coefficient of 0.9997 at levels near the detection limits up to at least 100 microg l(-1). The method was successfully applied to the determination of chromium in parenteral solution samples. PMID- 12560056 TI - Complete structure analysis of OR-1746, a complex product of cyclocondensation of arylhydrazomalononitriles containing clusters of protonated and unprotonated nitrogens, by pulsed-field-gradient heteronuclear NMR. AB - OR-1746, or [4-Ethoxy-6-imino-5-[[4-(4-methyl-6-oxo-1,4,5,6-tetrahydropyridazin-3 yl)phenyl]hydrazono]-5,6-dihydro-1H-pyrimidin-2-ylidene]-[4-(4-methyl-6-oxo 1,4,5,6-tetrahydropyridazin-3-yl)phenylazo]-acetonitrile is the product of the cyclocondensation of two molecules of the arylhydrazomalononitrile levosimendan (CAS registry number [141505-33-1]) with ethanol. OR-1746 is a molecule with a complex structure containing clusters of protonated and unprotonated nitrogens. Its structure was only partially elucidated by elemental analysis and by conventional NMR. However, the presence of many unprotonated nitrogen atoms did not allow the unambiguous assignment of the 1H, 13C and 15N NMR spectra with short-range heterocorrelated techniques, or even with traditional long-range 2D experiments. Pulsed-field-gradient heteronuclear multiple bond coherence sequences (PFG 1H-(13)C and PFG 1H-(15)N) were, therefore, used to fully assign the NMR spectra and elucidate the chemical structure of OR-1746. By using these techniques, long-range couplings between protons and carbons or proton and nitrogen atoms as distant as five bonds in the structure were detected without loosing the signals of the protonated heteroatoms. The long range coupling information provided by the novel NMR experiment can be used effectively in the complete structure determination of complex molecules containing clusters of protonated and unprotonated nitrogens. PMID- 12560057 TI - Optimisation for the separation of the oligosaccharide, sodium Pentosan Polysulfate by reverse polarity capillary zone electrophoresis using a central composite design. AB - The separation by reverse polarity capillary zone electrophoresis of the therapeutically developed sodium salt of Pentosan Polysulfate was optimised through the analysis of response surface methodologies, modeled using a central composite design. The optimisation investigated injection pressure, injection time and voltage and the effect of the conditions on retention times, peak areas, separation efficiency and the method sensitivity. The overall goal was to develop the most sensitive results with no decrease in separation efficiency. The following results were obtained: (1) retention times generally decreased as injection pressure, injection time and voltage increased, injection time having the least effect; (2) as expected peak areas increased as injection pressure and injection time increased but decreased as voltage increased; (3) separation efficiencies generally increased as injection pressure and injection time decreased, with voltage having almost no effect. For the optimum condition, the sample was introduced at the inlet vial at the cathode hydrodynamically, at optimal setting of 44 s at 35 mbar. The optimal voltage was -20 kV. In comparison with other methods, the optimum showed increased sensitivity, resolution and separation efficiency. Repeatability studies were performed on the optimum parameter conditions. Relative standard deviation values obtained were between 0.9 and 5.4%. PMID- 12560058 TI - Spectrophotometric determination of polyethylenimine in the presence of an oligonucleotide for the characterization of controlled release formulations. AB - Polyethylenimine (PEI) is a cationic polymer that can be associated to oligonuclotides to promote their transfection both in vitro and in vivo. The controlled release of oligonucleotide/polyethylenimine complexes from biodegradable systems can result in an increased cellular internalisation of the oligonucleotide and a reduced cytotoxicity of the complex. This effect strongly depends on the amount of PEI loaded in and released from the delivery system. In this work we describe a rapid, sensitive and reproducible spectrophotometric method for the quantitative analysis of PEI by itself or in the presence of an associated oligonucleotide. PEI does not possess chromophores, hence the determination by ordinary spectrophotometry is not possible. However, upon addition of copper (II) ions, PEI forms a dark blue cuprammonium complex that can be detected by UV-vis spectrophotometry. The optimum conditions in terms of optical parameters, copper (II) concentration required for a quantitative PEI complexation, and the most suitable medium for the reaction were ascertained. A linear relationship (r(2)=0.9997) between absorbance and amounts of PEI was found at lambda(max) of 285 nm over the concentration range 5.0-50.0 microg ml(-1). The detection limit (QOD) was 4.0 microg ml(-1). The method was validated for the quantitation of PEI in the presence of an oligonucleotide, which absorbs at 285 nm as well. PMID- 12560059 TI - Automated determination of selected water-soluble vitamins in tablets using a bench-top robotic system coupled to reversed-phase (RP-18) HPLC with UV detection. AB - The determination of folic acid, nicotinamide, nicotinic acid, riboflavin, riboflavin-5'-phosphate, pyridoxine and thiamine in tablets has been successfully automated using a bench-top robotic system (Zymark Tablet Processing Workstation II) coupled to reversed-phase (RP-18) HPLC with UV-detection. The new automated methods have been validated and were found to be applicable in routine analysis for all common types of multivitamin tablets. The precision is exemplified by relative standard deviations of 5.4% for folic acid, 2.1% for nicotinamide, 1.6% for pyridoxine, 0.7% for riboflavin and 1.1% for thiamine using film-coated tablets as sample. Mean recoveries obtained during spiking experiments were in the range of 95.2-103.9%. PMID- 12560060 TI - Prediction of drug release profiles using an intelligent learning system: an experimental study in transdermal iontophoresis. AB - This paper investigates the use of a neural-network-based intelligent learning system for the prediction of drug release profiles. An experimental study in transdermal iontophoresis (TI) is employed to evaluate the applicability of a particular neural network (NN) model, i.e. the Gaussian mixture model (GMM), in modeling and predicting drug release profiles. A number of tests are systematically designed using the face-centered central composite design (CCD) approach to examine the effects of various process variables simultaneously during the iontophoresis process. The GMM is then applied to model and predict the drug release profiles based on the data samples collected from the experiments. The GMM results are compared with those from multiple regression models. In addition, the bootstrap method is used to assess the reliability of the network predictions by estimating confidence intervals associated with the results. The results demonstrate that the combination of the face-centered CCD and GMM can be employed as a useful intelligent tool for the prediction of time series profiles in pharmaceutical and biomedical experiments. PMID- 12560061 TI - Determination of 5-aminosalicylic acid in pharmaceutical formulation by differential pulse voltammetry. AB - The oxidative behaviour of 5-aminosalicylic acid (5-ASA) has been investigated by differential pulse voltammetry using a glassy carbon electrode in different buffer systems. Linear sweep voltammetry was used to study the influence of pH on the peak current and peak potential. The solution conditions and instrumental parameters were optimized to obtain a good sensitivity. The Britton-Robinson buffer of pH 1.81 was selected as a suitable analytical medium in which 5-ASA exhibited a sensitive diffusion controlled oxidative peak at 0.564 V (vs. Ag/AgCl). The peak current varied linearly with drug concentration in the range between 1 x 10(-4) and 2 x 10(-6) M. The proposed voltammetric method has been applied to the determination of the drug in commercial delayed-release tablet forms. A mean recovery of 101.23% with a relative standard deviation of 1.35% was obtained. PMID- 12560062 TI - Analysis of 'SHENMAI' injection by HPLC/MS/MS. AB - An HPLC/MS/MS method was developed for the analysis of 'SHENMAI' injection, composed of red ginseng and ophiopogon. The constituents of 'SHENMAI' were found to be similar with those of ginseng and 39 ginsenosides were detected. By the studies of MS and MS/MS spectra and the comparison with literature data, most of these ginsenosides were identified. Based on this study, suggestions were put forward to improve the quality control system of 'SHENMAI' injection. PMID- 12560063 TI - Semi-micro high-performance liquid chromatographic analysis of tiropramide in human plasma using column-switching. AB - A rapid and sensitive column-switching semi-micro high-performance liquid chromatography method was developed for the direct analysis of tiropramide in human plasma. The plasma sample (100 microl) was directly injected onto Capcell Pak MF Ph-1 precolumn where deproteinization and analyte fractionation occurred. Tiropramide was then eluted into an enrichment column (Capcell Pak UG C(18)) using acetonitrile-potassium phosphate (pH 7.0, 50 mM) (12:88, v/v) and was analyzed on a semi-micro C(18) analytical column using acetonitrile-potassium phosphate (pH 7.0, 10 mM) (50:50, v/v). The method showed excellent sensitivity (limit of quantification 5 ng/ml), and good precision (C.V.Glu) is known to enhance tyrosine kinase activity, by promoting receptor dimerization. In order to gain insight into potential structural changes that arise as a result of the mutation, peptides corresponding to the complete transmembrane domain of proto-oncogenic and mutant forms of Neu have been studied by 1H nuclear magnetic resonance in the solvent trifluoroethanol (TFE). The chemical shifts are similar for both forms of the peptide, with the exception of amide residues close to the mutation site. Both peptides adopt a helical conformation, with a distinct bend one turn downstream of the mutation site. This deformation gives rise to several nuclear Overhauser effects, the majority of which were detected in both peptides, that are atypical for a straight canonical alpha-helix. Our data in this solvent do not support a conformational change in the transmembrane domain of monomeric Neu as a result of the mutation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicates that proto oncogenic Neu peptides have a higher propensity to oligomerize in the solvent TFE than the Glu664 oncogenic form. PMID- 12560076 TI - Mouse brain serine racemase catalyzes specific elimination of L-serine to pyruvate. AB - D-Serine was previously identified in mammalian brain and was shown to be a co agonist at the 'glycine' site of the N-methyl-D-aspartate (NMDA)-type receptors. Racemization of serine is catalyzed by serine racemase, a pyridoxal 5'-phosphate dependent enzyme expressed mainly in brain and liver. NMDA receptor overactivation has been implicated in a number of pathological conditions and inhibitors of serine racemase are thus potentially interesting targets for therapy. We expressed recombinant mouse serine racemase in insect cells and purified it to near homogeneity. The enzyme is a non-covalent homodimer in solution and requires divalent cations Mg(2+), Ca(2+) or Mn(2+) for activity but not for dimerization. In addition to the racemization it also catalyzes specific elimination of L-Ser to pyruvate. D-Serine is eliminated much less efficiently. Both L-serine racemization and elimination activities of serine racemase are of comparable magnitude, display alkaline pH optimum and are negligible below pH 6.5. PMID- 12560077 TI - Phosphorylation of serine residues is fundamental for the calcium-binding ability of Orchestin, a soluble matrix protein from crustacean calcium storage structures. AB - Orchestia cavimana is a terrestrial crustacean, which cyclically stores calcium in diverticula of the midgut, in the form of calcified amorphous concretions. These concretions are associated with a proteinaceous matrix, the main constituent of the soluble matrix is Orchestin, an acidic calcium-binding protein [Testeniere et al., Biochem. J. 361 (2002) 327-335]. In the present paper, we clearly demonstrate that Orchestin is phosphorylated on serine and tyrosine residues, but that calcium binding only occurs via the phosphoserine residues. To our knowledge, this is the first example of an invertebrate mineralization for which a post-translational modification is clearly related to an important function of a calcifying protein. PMID- 12560078 TI - The elasticity of single kettin molecules using a two-bead laser-tweezers assay. AB - Kettin is a high molecular mass protein of insect muscle associated with thin filaments and alpha-actinin in the Z-disc. It is thought to form a link between thin and thick filaments towards its C-terminus, contributing significantly to passive sarcomere stiffness. Here the elastic properties were characterised by mechanical stretches on an antibody-delimited region of the single molecule using two independent optical traps capable of exerting forces up to 150 pN. Step-like events were observed in the force-extension relationships consistent with the unfolding of Ig domains at moderate force and refolding of these domains at significantly higher forces than have been observed for related modular proteins. PMID- 12560079 TI - Identification and characterization of Mlr1,2: two mouse homologues of Mblk-1, a transcription factor from the honeybee brain(1). AB - We previously identified the Mblk-1 gene in the honeybee brain, which encodes a transcription factor containing two DNA binding motifs, termed RHF1 and 2 (Takeuchi et al. (2001) Insect Mol. Biol. 121, 134-140). Here, we identified two mouse Mblk1 homologues, Mlr1 and Mlr2. Both encode proteins containing a single DNA-binding motif highly conserved with RHF2 and activate transcription mediated by a DNA element recognized by honeybee Mblk-1. Mlr1 was expressed predominantly in the spermatocytes of the testis, while Mlr2 was expressed in various tissues other than testis. Mlr1 transcripts were lost in the testis of W/W(v) mutant mice, suggesting a role in spermatogenesis. PMID- 12560080 TI - Interaction of the disordered terminal regions of flagellin upon flagellar filament formation. AB - Helical filaments of bacterial flagella are built up by a self-assembly process from thousands of flagellin subunits. To clarify how the disordered terminal regions of flagellin interact upon filament formation, polymerization ability of various terminally truncated fragments was investigated. Fragments deprived of 19 N-terminal residues were able to bind to the end of filaments, however, only a single layer was formed. Removal of C-terminal segments or truncation at both ends resulted in the complete loss of binding ability. Our observations are consistent with the coiled-coil model of filament formation, which suggests that the alpha-helical N- and C-terminal regions of axially adjacent subunits form an interlocking pattern of helical bundles upon polymerization. PMID- 12560081 TI - Modulation of EGF receptor autophosphorylation by alpha-hemolysin of Staphylococcus aureus via protein tyrosine phosphatase. AB - In the presence of assembled alpha-hemolysin (alpha-HL) of Staphylococcus aureus, the epidermal growth factor receptor (EGFr) is rapidly dephosphorylated. Several obvious possibilities that otherwise would have contributed to the dephosphorylation were ruled out. Instead, an elevation in the activity of a protein tyrosine phosphatase appears to be responsible for the observed loss of phosphorylation signal of EGFr. For this dephosphorylation, the assembly of alpha HL is necessary while lytic pore formation is not required. In summary, the EGFr is unable to retain its phosphorylation signal in the presence of alpha-HL and the process is irreversible. PMID- 12560082 TI - Ubiquitin binding proteins protect ubiquitin conjugates from disassembly. AB - As a step in their turnover proteins in eukaryotic cells are coupled to a small protein, ubiquitin, before they are recognised by 26S proteasomes and degraded. However, cells also contain many deubiquitinating enzymes, which can rescue proteins by cleaving off the ubiquitin chains. Here we report that three ubiquitin binding proteins, Rhp23, Dph1 and Pus1, from fission yeast can protect multiubiquitin conjugates against deubiquitination. This protection depends on the ubiquitin binding domains and may promote degradation of ubiquitinated proteins. PMID- 12560083 TI - Ezrin is a substrate for Lck in T cells. AB - We evaluated the role of Lck tyrosine kinase, an early effector of T cell activation, in regulation of the membrane-cytoskeleton linker protein ezrin. Ezrin was constitutively tyrosine phosphorylated in wild-type and CD45-deficient Jurkat T cells, but not in Lck-deficient cells. However, phosphorylation was evident in cells, in which Lck activity had been restored by transfection. Phosphorylation was reduced by the Src family kinase inhibitor PP2 and increased by the tyrosine phosphatase inhibitor pervanadate, implying continuous tyrosine phosphorylation and dephosphorylation. Lck phosphorylated ezrin in vitro, and the major phosphotyrosine was identified as Y145. These results identify ezrin as the first cytoskeletal substrate for Lck. PMID- 12560084 TI - Fluid shear stress-induced transcriptional activation of the vascular endothelial growth factor receptor-2 gene requires Sp1-dependent DNA binding. AB - Hemodynamic forces play a fundamental role in the regulation of endothelial cell survival. As signaling via the vascular endothelial growth factor (VEGF) receptor 2 pathway has been previously demonstrated to impact endothelial cell survival, we hypothesized that laminar shear stress may facilitate survival in part by inducing VEGF receptor-2 expression. This study shows a time- and dose-dependent upregulation of endothelial VEGF receptor-2 expression by fluid shear stress in microvascular and large-vessel derived endothelial cells. A functional analysis of the 5'-regulatory region of the VEGF receptor-2 promoter localized the shear stress-response element to a sequence between bp -60 and -37 that encompasses two adjacent consensus Sp1 transcription factor binding sites. Constitutive and shear stress-inducible Sp1-dependent complexes are bound to this element, indicating that fluid shear stress-induced transcriptional activation of the VEGF receptor-2 gene requires Sp1-dependent DNA binding. Together, these results suggest that biomechanical stimulation may lead to endothelial cell survival by upregulating VEGF receptor-2 expression. PMID- 12560085 TI - Solution structure of a tmRNA-binding protein, SmpB, from Thermus thermophilus. AB - Small protein B (SmpB) is required for trans-translation, binding specifically to tmRNA. We show here the solution structure of SmpB from an extremely thermophilic bacterium, Thermus thermophilus HB8, determined by heteronuclear nuclear magnetic resonance methods. The core of the protein consists of an antiparallel beta barrel twisted up from eight beta-strands, each end of which is capped with the second or third helix, and the first helix is located beside the barrel. Its C terminal sequence (20 residues), which is rich in basic residues, shows a poorly structured form, as often seen in isolated ribosomal proteins. The results are discussed in relation to the oligonucleotide binding fold. PMID- 12560086 TI - Sphingosine-1-phosphate stimulates cortisol secretion. AB - We show here for the first time that sphingosine-1-phosphate (Sph-1-P) stimulates cortisol secretion in zona fasciculata cells of bovine adrenal glands. This effect was dependent upon protein kinase C (PKC) and extracellular Ca2+, and was inhibited by pertussis toxin. Sph-1-P activated phospholipase D (PLD) through a pertussis toxin-sensitive mechanism, also involving extracellular Ca2+ and PKC. Primary alcohols, which attenuate formation of phosphatidic acid (the product of PLD), and cell-permeable ceramides, which inhibit PLD, blocked Sph-1-P-induced cortisol secretion. In conclusion, Sph-1-P stimulates cortisol secretion through a mechanism involving Gi/o protein-coupled receptors, extracellular Ca2+, PKC and PLD. PMID- 12560087 TI - HIF-1 alpha protein as a target for S-nitrosation. AB - Hypoxia-inducible factor-1 alpha (HIF-1 alpha) is a master regulator to sense decreased oxygen partial pressure. HIF-1 alpha stability regulation initiates a complex biological response that allows cells to act appropriately to meet patho physiological situations of decreased oxygen availability. Recently, nitric oxide emerged as a messenger with the ability to stabilize HIF-1 alpha and to transactivate HIF-1 under normoxia. Considering that reactive nitrogen species are recognized for post-translation protein modifications, among others S nitrosation, we asked whether HIF-1 alpha is a target for S-nitrosation. In vitro NO+ donating NO donors such as GSNO and SNAP provoked massive S-nitrosation of purified HIF-1 alpha. All 15 free thiol groups found in human HIF-1 alpha are subjected to S-nitrosation. Thiol modification is not shared by spermine-NONOate, a NO radical donating compound. However, spermine-NONOate in the presence of O(2)(-), generated by xanthine/xanthine oxidase, regained S-nitrosation, most likely via formation of a N(2)O(3)-like species. In vitro, S-nitrosation of HIF-1 alpha was attenuated by the addition of GSH or ascorbate. In RCC4 and HEK293 cells GSNO or SNAP reproduced S-nitrosation of HIF-1 alpha, however with a significantly reduced potency that amounted to modification of three to four thiols, only. Importantly, endogenous formation of NO in RCC4 cells via inducible NO synthase elicited S-nitrosation of HIF-1 alpha that was sensitive to inhibition of inducible NO synthase activity with N-monomethyl-L-arginine. NO stabilized HIF-1 alpha was susceptible to the addition of N-acetyl-cysteine that destabilized HIF-1 alpha in close correlation to the disappearance of S nitrosated HIF-1 alpha. In conclusion, HIF-1 alpha is a target for S-nitrosation by exogenously and endogenously produced NO. PMID- 12560088 TI - Slow and persistent increase of [Ca2+]c in response to ligation of surface IgM in WEHI-231 cells. AB - WEHI-231 and Bal 17 B cell lines are representative models for immature and mature B cells, respectively. Their regulation of cytosolic Ca(2+) concentration ([Ca(2+)](c)) was compared using fura-2 fluorescence ratiometry. The ligation of B cell antigen receptor (BCR) by anti-IgM antibody induced a slow but large increase of [Ca(2+)](c) in WEHI-231 cells while not in Bal 17 cells. The thapsigargin-induced store-operated Ca(2+) entry (SOCE) of Bal 17 cells reached a steady state which was blocked by 2-aminoethoxydiphenyl borate (2-APB). On the contrary, the thapsigargin-induced SOCE of WEHI-231 cells increased continuously, which was accelerated by 2-APB. The increase of [Ca(2+)](c) by BCR ligation was also enhanced by 2-APB in WEHI-231 cells while blocked in Bal 17 cells. The Mn(2+) quenching study showed that the thapsigargin-, or the BCR ligation-induced Ca(2+) influx pathway of WEHI-231 was hardly permeable to Mn(2+). The intractable increase of [Ca(2+)](c) may explain the mechanism of BCR-driven apoptosis of WEHI 231 cells, a well-known model of clonal deletion of autoreactive immature B cells. PMID- 12560089 TI - Inhibition of the yeast V-type ATPase by cytosolic ADP. AB - The activity of the vacuolar H(+)-ATPase has been characterized in isolated vacuoles of the yeast Saccharomyces cerevisiae by means of the patch-clamp technique. With cytosolic calcium at virtually zero (<10(-9) M), Mg-ATP induced a transient, bafilomycin A(1)-sensitive current corresponding to the flow of positive charges from the cytoplasmic surface to the vacuolar lumen. The Mg-ATP dependent current reached its maximum amplitude (30+/-8 mA m(-2) with 5 mM Mg ATP, n=34) within 15-20 s and declined slowly over a period of about 15-20 min even in the continuous presence of Mg-ATP. This decline of pumping activity was independent of the cytosolic KCl concentration, suggesting an inhibitory mechanism different from the high salt-induced dissociation of V(0) and V(1) reported for the V-ATPase of plants and fungi. Cytosolic ADP was found to modulate the pump activity since Mg-ATP-induced pump current was smaller if monitored in the presence of 5 mM ADP and addition of 5 mM ADP in the presence of 5 mM Mg-ATP reduced the pump current by more than 50%. Furthermore, reduction of the cytosolic ADP concentration by the ATP-regenerating system creatine phosphate/creatine kinase partially relieved the endogenous inhibition of the V ATPase, confirming that interaction of cytosolic ADP with the V-ATPase is the reason for the transient nature of the pump current in yeast vacuoles. PMID- 12560090 TI - Relevance of the proximal domain in the amino-terminus of HERG channels for regulation by a phospholipase C-coupled hormone receptor. AB - We used Xenopus oocytes co-expressing thyrotropin-releasing hormone (TRH) receptors and human ether-a-go-go-related gene (HERG) K+ channel variants carrying different amino-terminal modifications to check the relevance of the proximal domain for hormonal regulation of the channel. Deletion of the whole proximal domain (Delta 138-373) eliminates TRH-induced modifications in activation and deactivation parameters. TRH effects on activation are also suppressed with channels lacking the second half of the proximal domain or only residues 326-373. However, normal responses to TRH are obtained with Delta 346 373 channels. Thus, whereas residues 326-345 are required for the hormonal modulation of HERG activation, different proximal domain sequences contribute to set HERG gating characteristics and its regulation by TRH. PMID- 12560091 TI - Candidate markers for stem and early progenitor cells, Musashi-1 and Hes1, are expressed in crypt base columnar cells of mouse small intestine. AB - Musashi-1, a neural RNA-binding protein, is important for maintaining neural stem cells. Both Musashi-1 and Hes1, a transcriptional factor regulated by Musashi-1, are expressed in the small intestine. Here we show that Musashi-1 is present in a few epithelial cells just above the Paneth cells in the small intestinal crypt, the putative position of stem cells, whereas Hes1 is expressed in lower crypt cells just above the Paneth cells, including Musashi-1-positive cells. Musashi-1 and Hes1 were not expressed in Paneth cells. Notably, Musashi-1 and Hes1 were coexpressed in the crypt base columnar cells located between the Paneth cells. These findings suggest that not only the cells just above Paneth cells but also the crypt base columnar cells between the Paneth cells have stem cell characteristics. PMID- 12560092 TI - In plants the alc gene expression system responds more rapidly following induction with acetaldehyde than with ethanol. AB - It has recently been proposed that acetaldehyde is the physiological inducer of the alc gene system and hence indirectly the activator of the AlcA promoter in Aspergillus nidulans. Here we show that this chemical induces expression of a GUS (beta-D-glucuronidase) reporter under the control of the alc gene system in transgenic potato tubers more rapidly than ethanol allowing tighter control of transgene expression. Furthermore by analysis of metabolite levels we demonstrate that the application of inducer has few effects on metabolism. We propose that this system is therefore ideal for the temporal regulation of important metabolic enzyme activities. PMID- 12560093 TI - Apparent affinity of CFTR for ATP is increased by continuous kinase activity. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel which is activated by protein phosphorylation and nucleoside triphosphates. We demonstrate here that fusion of the soluble catalytic subunit of cAMP-dependent protein kinase to the membrane protein bacteriorhodopsin yields a constitutively active protein kinase which activates CFTR effectively. As it is membrane-bound it is particularly useful for continuous perfusion of excised inside-out patches. We also tested the effect of a naturally membrane-bound protein kinase, cGMP-dependent protein kinase II, on CFTR. Both kinases, when continuously active, increase apparent affinity of CFTR to ATP about two-fold emphasizing the role of phosphorylation in modulating the interaction of ATP with the nucleotide binding domains. PMID- 12560094 TI - Interplay between hnRNP A1 and a cis-acting element in the 3' UTR of CYP2A5 mRNA is central for high expression of the gene. AB - Our previous evidence suggests that heterogeneous nuclear ribonucleoprotein (hnRNP) A1 plays a part in the regulation of the Cyp2a5 gene by interacting with the 3' untranslated region (UTR) of the CYP2A5 mRNA. However, the exact role of this interaction is not clear. The aim of the present work was to gain further insight into the regulation process of Cyp2a5. For this purpose the 3' UTR of CYP2A5 was fused to the coding region of luciferase mRNA. Luciferase recombinants containing either the full length 3' UTR, or the 3' UTR lacking a previously described 71 nucleotide (nt) region (the hnRNP A1 primary binding site), were transiently expressed in cells expressing or lacking hnRNP A1. The expression of the luciferase recombinants was examined both at mRNA and enzyme activity levels. The results disclosed that the presence of hnRNP A1 was required for the high expression of the recombinant carrying the full length 3' UTR of CYP2A5. Deletion of the hnRNP A1 primary binding site dramatically modified the expression pattern: the mRNA levels and luciferase activities of the deletion mutant were independent from hnRNP A1. These results conclusively demonstrate that the 71 nt region in the 3' UTR of CYP2A5 mRNA can confer hnRNP A1-dependent regulation to a gene. In addition, comparison of RNA levels and luciferase activities suggested that regions flanking the hnRNP A1 binding site could regulate translation of the CYP2A5 mRNA. These results are consistent with a model in which the binding of hnRNP A1 to the 71 nt putative hairpin-loop region in the CYP2A5 mRNA 3' UTR upregulates mRNA levels possibly by protecting the mRNA from degradation. PMID- 12560095 TI - 17 beta-Oestradiol attenuates nucleotide excision repair. AB - Epidemiological studies strongly suggest associations between chronic exposure to endogenous oestrogens and the development of breast and gynaecological tumours. Two mechanisms by which 17 beta-oestradiol (E2) may enhance tumorigenesis are: (i) enhancement of cell proliferation and (ii) the production of reactive, genotoxic metabolites. Here we suggest an additional mechanism, inhibition of DNA repair. The removal of UV-induced thymine dimers from human keratinocytes, reflective of nucleotide excision repair, was significantly attenuated by treatment of cells with E2. In contrast, treatment with 17 alpha-oestradiol had no effect. Mechanisms are proposed for this effect of E2, which may contribute to its carcinogenic potential. PMID- 12560096 TI - Mossbauer studies of the non-heme iron and cytochrome b559 in a Chlamydomonas reinhardtii PSI- mutant and their interactions with alpha-tocopherol quinone. AB - Spin and valence states of the non-heme iron and the heme iron of cytochrome b559, as well as their interactions with alpha-tocopherol quinone (alpha-TQ) in photosystem II (PSII) thylakoid membranes prepared from the Chlamydomonas reinhardtii PSI- mutant have been studied using Mossbauer spectroscopy. Both of the iron atoms are in low spin ferrous states. The Debye temperature of the non heme is 194 K and of the heme iron is 182 K. The treatment of alpha-TQ does not change the spin and the valence states of the non-heme iron but enhances the covalence of its bonds. alpha-TQ oxidizes the heme iron into the high spin Fe3+ state. A possible role of the non-heme iron and alpha-TQ in electron flow through the PSII is discussed. PMID- 12560097 TI - Sequence analysis reveals new membrane anchor of reaction centre-bound cytochromes possibly related to PufX. AB - Most of the bacterial photosynthetic reaction centres known to date contain a cytochrome subunit with four covalently bound haem groups. In the case of Blastochloris viridis, this reaction centre subunit is anchored in the membrane by a lipid molecule covalently attached to the cysteine which forms the N terminus of the mature protein after processing by a signal peptidase. We show that posttranslational N-terminal cleavage of the cytochrome subunit does not occur in the aerobic photosynthetic bacterium Roseobacter denitrificans. From sequence analysis of the resulting elongated N-terminus it follows that a transmembrane helix is anchoring the reaction centre-bound cytochrome in the membrane. Comparative sequence analysis strongly suggests that all cytochrome subunits lacking the lipid coupling cysteine share this structural feature. Comparison of the N-terminal segment of the cytochrome subunit of Roseobacter denitrificans with the sequences of the PufX proteins from Rhodobacter sphaeroides and Rhodobacter capsulatus suggests a phylogenetic relation. PMID- 12560098 TI - Characterization of a novel Drosophila melanogaster acylphosphatase. AB - Analysis of the Drosophila melanogaster EST database led to the characterization of a novel acylphosphatase (AcPDro2). This is coded by the CG18505 (Acyp2) gene and is clearly distinct from a previously described AcPDro coded by the CG16870 (Acyp) gene from D. melanogaster. The two proteins show a 60% homology with both vertebrate isoenzymes. All the residues involved in the catalytic mechanism are conserved. AcPDro2 is a stable enzyme with a correct globular folded structure. Its activity on benzoylphosphate shows higher K(cat) but lower K(m) with respect to AcPDro. It is possible that AcPDro and AcPDro2 genes are not the direct ancestor of MT and CT vertebrate isoenzymes. PMID- 12560099 TI - Proteolytic stability of beta-peptide bonds probed using quenched fluorescent substrates incorporating a hemoglobin cleavage site. AB - A set of designed internally quenched fluorescence peptide substrates has been used to probe the effects of insertion of beta-peptide bonds into peptide sequences. The test sequence chosen corresponds to a proteolytically susceptible site in hemoglobin alpha-chain, residues 32-37. Fluorescence and mass spectral measurements demonstrate that the insertion of an beta-residues at the potential cleavage sites completely abolishes the action of proteases; in addition, the rate of cleavage of the peptide bond preceding the site of modification is also considerably reduced. PMID- 12560101 TI - Stress-induced hyperphosphorylation of tau in the mouse brain. AB - We previously showed that starvation causes reversible hyperphosphorylation of tau in the mouse brain. To explore possible involvement of stress in tau hyperphosphorylation quantitative analysis of phosphorylated tau in four brain regions of mice subjected to cold water stress (CWS) was made by immunoblot analyses using phosphorylation-dependent antibodies directed to eight sites on tau known to be hyperphosphorylated in the brain of Alzheimer's disease (AD) patients. Ser199, Ser202/Thr205, Thr231/Ser235 were hyperphosphorylated 20 and 40 min after CWS. The response was pronounced in the hippocampus and cerebral hemisphere, but weak in the cerebellum in parallel with the regional vulnerability in AD. Among the regulatory phosphorylation of protein kinases studied, a transient phosphorylation of tau protein kinase I/glycogen synthase kinase 3beta at Ser9 was most conspicuous. PMID- 12560100 TI - Redox catalysts as sensitisers towards oxidative stress. AB - The predominance of oxidative stress in many tumour cell environments provides a means to selectively target these cells via protein oxidation. The zinc fingers of transcription factors utilise cysteine thiols for structural zinc coordination. Redox control of DNA binding regulates transcription and therefore the overall rates of proliferation, apoptosis and necrosis in the carcinoma. We report here the adverse effects of glutathione peroxidase (GPx) mimics towards zinc finger motifs and PC12 cell survival. Nanomolar catalyst concentrations facilitated H2O2-induced oxidation of an Sp1 transcription factor fragment. In PC12 cells GPx catalysis triggered a significant increase in cell death, correlating with severity of oxidative stress. As a consequence, we conclude that GPx mimics are potential chemotherapeutic agents. PMID- 12560102 TI - Enhancement of TNF-alpha-mediated cell death in vascular smooth muscle cells through cytochrome c-independent pathway by the proteasome inhibitor. AB - There is substantial evidence that cytokines induce apoptosis of vascular smooth muscle cells (VSMCs) in atherosclerosis. Its regulation, however, is not completely defined. The aim of this study is to investigate whether proteasome activity is related with apoptosis in VSMCs by tumor necrosis factor-alpha (TNF alpha). Rat aorta smooth muscle cells were treated with TNF-alpha and proteasome inhibitor MG132 and then cell death was determined by morphology, viability, and DNA fragmentation. MG132 or TNF-alpha alone did not induce cell death. In contrast, co-treatment of TNF-alpha and proteasome inhibitor induced death and DNA degradation in VSMCs, suggesting proteasome inhibitor enhanced death activity of TNF-alpha. The death was not blocked by ascorbic acid but by nitric oxide synthase inhibitor N(G)-monomethyl-L-arginine. Both caspase-3 and -8 were activated during the death by the proteasome inhibitor and TNF-alpha. The death was effectively blocked by the caspase-3 inhibitor z-DEVD-fmk, suggesting a role of caspase-3 in the death. Nonetheless, there were no significant alterations in the level of Bcl-2, Bcl-X(L), Bax and Bak by the proteasome inhibitor, nor any evidence of cytochrome (cyt) c release into cytosol from dying cells, suggesting that cyt c is not involved. These results suggest that proteasome inhibition potentiates TNF-mediated death in VSMCs in a cyt c-independent pathway. The present study proposes a new mechanism by which VSMCs undergo death by cytokines. PMID- 12560103 TI - Antibacterial activity of peptides derived from envelope glycoproteins of HIV-1. AB - Recent reports have highlighted the anti-HIV-1 activities of defensins, whose structure and charge resemble portions of the HIV-1 transmembrane envelope glycoprotein gp41. The current report explores the obverse, whether peptides derived from HIV-1 envelope glycoproteins can exert antimicrobial activity. Fifteen-residue peptides spanning the entire sequence of HIV-1(MN) gp120 and gp41 were subjected to radial diffusion assays against laboratory strains of Escherichia coli and Listeria monocytogenes. Twenty-four active peptides corresponded predominantly to membrane-active domains of gp120 and gp41. Several peptides retained significant activity in higher ionic conditions and may serve as templates for the development of novel peptide antibiotics. The strategies employed herein could uncover additional antimicrobial peptides from envelope proteins of other lytic viruses. PMID- 12560104 TI - Characterization of the cupin-type phosphoglucose isomerase from the hyperthermophilic archaeon Thermococcus litoralis. AB - The gene encoding phosphoglucose isomerase was cloned from Thermococcus litoralis, and functionally expressed in Escherichia coli. The purified enzyme, a homodimer of 21.5 kDa subunits, was biochemically characterized. The inhibition constants for four competitive inhibitors were determined. The enzyme contained 1.25 mol Fe and 0.24 mol Zn per dimer. The activity was enhanced by the addition of Fe(2+), but inhibited by Zn(2+) and EDTA. Enzymes with mutations in conserved histidine and glutamate residues in their cupin motifs contained no metals, and showed large decreases in k(cat). The circular dichroism spectra of the mutant enzymes and the wild type enzyme were essentially the same but with slight differences. PMID- 12560105 TI - Alternative splicing of myeloid IgA Fc receptor (Fc alpha R, CD89) transcripts in inflammatory responses. AB - More than 10 splice variants of the Fc receptor for IgA (Fc alpha R, CD89) have been identified in human myeloid cells. In this study, we quantified Fc alpha R splice transcripts Delta EC2 and Delta 66 EC2, which lack the entire and a part of the homologous immunoglobulin-like extracellular domain 2 (EC2), respectively. Tumor necrosis factor-alpha was found to specifically increase the ratio of Delta EC2 to the wild type CD89 in neutrophils and conversely decrease the Delta EC2 ratio in monocytes. We also observed a significant decrease in the neutrophil Delta EC2/CD89 ratio in pneumonia patients. These results suggest that Delta EC2 is differentially regulated and could be involved in immunoregulation of IgA mediated host defense. PMID- 12560106 TI - D-2-amino-5-phosphonopentanoic acid inhibits intrathecal pertussis toxin-induced thermal hyperalgesia and protein kinase Cgamma up-regulation. AB - The aim of the present study was to examine the effect of intrathecal (i.t.) injection of pertussis toxin (PTX) on the nociceptive threshold and protein kinase C (PKC) expression in the rat spinal cord. The role of N-methyl-D-aspartic acid (NMDA) receptors in these changes was also examined. Male Wistar rats were implanted with two i.t. catheters, one of which was connected to a mini-osmotic pump and used to infuse saline or D-2-amino-5-phosphonopentanoic acid (D-AP5) (2 microg/h) starting on day 3 after i.t. catheter insertion. Two days later, a single injection of saline or PTX (2 microg) was given via the other catheter, followed by a flush with 10 microl of saline. On day 4 after PTX or saline injection, the thermal paw withdrawal latency was measured, then the rats were sacrificed by decapitation, and the dorsal part of the lumbosacral spinal segments was removed for PKC Western blotting assays. In PTX-treated rats, thermal hyperalgesia was observed, and the PKCgamma content of both the synaptosomal membrane and cytosolic fractions was significantly increased. The levels of alpha-, betaI-, or betaII-PKC isozymes in these fractions were unaffected by PTX treatment. Infusion of the NMDA antagonist, D-AP5, prevented both the thermal hyperalgesia and the increase in PKCgamma isoform expression in PTX-treated rats, and had no effect on these values in nai;ve rats. Intrathecal injection of the PKC inhibitor, chelerythrine (10 microg), significantly inhibited the thermal hyperalgesia observed in PTX-treated rats. These results show that i.t. injection of PTX induced thermal hyperalgesia accompanied by a selective increase in PKCgamma expression in both the synaptosomal membrane and cytosolic fractions of the dorsal horn of the rat lumbar spinal cord, and both effects were inhibited by the NMDA receptor antagonist, D-AP5. PMID- 12560107 TI - Protective effect of a new nonpeptidyl mimetic of SOD, M40401, against focal cerebral ischemia in the rat. AB - We tested the neuroprotective effects of M40401, a new, low molecular weight (511.4 Da) maganese superoxide dismutase mimetic, against 90 min of middle cerebral artery occlusion (MCAO) in male Wistar rats. Animals received a single injection of vehicle (n=8), 1 mg/kg (n=6), or 3 mg/kg (n=7) 30 min before MCAO. Total lesion volume was reduced only in the group receiving 3 mg/kg M40401 (163.5+/-18.7 versus 43.4+/-7.0 mm(3), for vehicle and M40401, respectively; P<0.05), with almost complete reduction of lesion volume in the cortex but little protection in the basal ganglia. Neurological score was also improved in this group. The dose of 1 mg/kg M40401 had smaller and inconsistent effects on lesion parameters. Administration of a single dose of 3 mg/kg M40401 at 60 min of MCAO or at the end of MCAO (90 min) failed to significantly reduce lesion volume. A single dose of M40401 plus prolonged infusion into the post-MCAO period also failed to decrease lesion volume significantly. These data indicate that M40401 protects cerebral tissue from ischemic insult when administered before MCAO, probably by limiting damage mediated by detrimental actions of superoxide anion. PMID- 12560108 TI - Cannabinoid-induced Fos expression within A10 dopaminergic neurons. AB - Mesocorticolimbic dopaminergic systems subserve cognitive processes, motivated behavior, the central stress response, and the reinforcing properties of drugs of abuse. Hyperdopaminergic states have been suggested to contribute to the psychotropic effects of the cannabinoids; however, the mechanisms by which cannabinoids activate mesocorticolimbic dopaminergic systems are not well understood. We have examined the role of noradrenergic neurotransmission in the mediation of cannabinoid-induced activation of A10 dopaminergic neurons using Fos as a marker of neuronal activation in mice. Administration of the CB(1) receptor agonist CP55940 differentially increased the number of Fos-like immunoreactive (Fos-li) A10 dopaminergic cells within three anatomically distinct regions (parabrachial pigmented, paranigral, and caudal linear nuclei) compared to vehicle-treated mice. Similar results were obtained using the CB(1) receptor agonist Win 55212-2; and pretreatment with the CB(1) receptor antagonist SR141716 significantly inhibited CP55940-induced Fos expression. Pretreatment with the alpha(1)-adrenergic receptor antagonist, prazosin, and the alpha(2)-adrenergic receptor agonist, clonidine, reduced the number of Fos-li dopaminergic neurons induced by CP55940 in a subregion-specific manner. CP55940 and Win 55212-2 increased the number of Fos-li neurons within the locus coeruleus. Finally, CB(1) receptor immunoreactivity was detected on fibers within the CL but not in either PBP or PN. Our data demonstrate that cannabinoids induce Fos expression within A10 dopaminergic neurons in a heterogeneous anatomical pattern, and suggest that enhanced noradrenergic neurotransmission contributes to cannabinoid-induced activation of A10 dopaminergic neurons in vivo. PMID- 12560109 TI - Direct injection of substance P-antisense oligonucleotide into the feline NTS modifies the cardiovascular responses to ergoreceptor but not baroreceptor afferent input. AB - Substance P (SP) is released from the feline nucleus tractus solitarius (NTS) in response to activation of skeletal muscle afferent input. However, there are differing results about SP release from the rostral NTS in response to baroreceptor afferent input. An anti-sense oligonucleotide to feline SP (SP asODN) was injected directly into the rostral NTS of chloralose-anesthetized cats to determine whether blood pressure or heart rate responses to ergoreceptor activation (muscle contraction) or baroreceptor unloading (carotid artery occlusion) were sensitive to SP knockdown. Control injections included either buffer alone or a scrambled-sequenced oligonucleotide (SP-sODN). Both muscle contractions and carotid occlusions were performed 3, 6 and 12 h after the completion of the oligonucleotide injections. The cardiovascular responses to contractions were significantly attenuated 3 and 6 h after SP-asODN, but not by the injection of the SP-sODN. The cardiovascular responses to contractions returned to control levels 12 h post anti-sense injection. No detectable release of SP (using antibody-coated microprobes) was measured 3 and 6 h after SP-asODN injections and the expression of SP-immunoreactivity (SP-IR) in the NTS was significantly attenuated, as determined by immunohistochemistry procedures. In contrast, neither the injection of SP-asODN nor the s-ODN attenuated the cardiovascular responses to carotid occlusions, or altered the pattern of release of SP from the brainstem. Injection of the SP-sODN did not affect the expression of SP-IR. These results suggest that the SP involved with mediating the peripheral somatomotor signal input to the rostral NTS comes from SP-containing neurons within the NTS. Our results also suggest that SP in the rostral NTS does not play a direct role in mediating the cardiovascular responses to unloading the carotid baroreceptors. We suggest that the SP released during isometric contractions excites an inhibitory pathway modulating baroreceptor input, thus contributing to the increase in mean blood pressure. PMID- 12560111 TI - Glutamatergic afferent projections to the dorsal raphe nucleus of the rat. AB - Based on WGA-apo-HRP-gold (WG) retrograde tracing, the present study revealed that different subdivisions of the dorsal raphe (DR) such as dorsomedial, ventromedial, lateral wing, and caudal regions receive unique, topographically organized afferent inputs, that are more restricted than previously reported. Phaseolus vulgaris leucoagglutinin anterograde tracing studies confirmed that the medial prefrontal cortex provides the major afferent input to each subdivision of the DR. Double-labeling studies combining WG tracing and glutamate immunostaining indicated that the medial prefrontal cortex, various hypothalamic nuclei including perifornical, lateral, and arcuate nuclei, and several medullary regions such as lateral and medial parabrachial nuclei, and laterodorsal tegmental nucleus provide the major glutamatergic input to each subregion of the DR. It should be noted that the degree of glutamatergic input from these afferent sites was specific for each DR subdivision. The present findings indicated that dorsomedial, ventromedial, lateral wing, and caudal subdivisions of the DR receive excitatory inputs from both cortical and subcortical sites which might be involved in regulation or modulation of a broad range of systems, including sensory and motor functions, arousal and sleep-wake cycle, biorhythmic, cognitive, and affective behaviors. PMID- 12560110 TI - CI-1010 induced opening of the mitochondrial permeability transition pore precedes oxidative stress and apoptosis in SY5Y neuroblastoma cells. AB - The hetero-bifunctional nitroimidazole radiosensitizer CI-1010, R-alpha-[[(2 bromoethyl)-amino]methyl]-2-nitro-1H-imidazole-1-ethanol monohydrobromide, causes selective irreversible apoptotic loss of retinal photoreceptor cells in vivo. The human neuroblastoma cell line, SH-SY5Y, was used as a neuronotypic model of CI 1010-mediated retinal degeneration. Exposure to CI-1010 for 24 h induced apoptosis in neuroblastoma cells, as determined by histopathological and ultrastructural analysis and by TUNEL technique. CI-1010 causes a dose-dependent decrease in cell viability in SY5Y cells, as measured by the reduction of MTT, 3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. Superoxide dismutase reduced loss of cell viability following CI-1010 treatment suggesting an oxidative stress-mediated mechanism of toxicity. The effects of CI-1010 on mitochondrial membrane potential and intracellular levels of reactive oxygen species were assessed in live SY5Y cells by confocal microscopy using the fluorescent dyes, tetramethylrhodamine methyl ester and 5,6-carboxy-2',7' dihydrodichlorofluorescein diacetate. CI-1010 caused a rapid depolarization of mitochondria in SY5Y cells followed by an increase in ROS. Both CI-1010-induced mitochondrial depolarization and subsequent increases in ROS were prevented by pretreatment with either the permeability transition pore inhibitor, cyclosporin A (CsA), and by the antioxidant, alpha-tocopherol. However, CsA and alpha tocopherol were unable to prevent apoptosis in CI-1010-treated cells, suggesting the influence of additional mechanism(s) of CI-1010-induced toxicity. This study evaluates intracellular oxidative stress associated with pore opening prior to apoptosis and provides evidence in support of a mitochondrial mechanism of CI 1010-induced neuronal cell death. PMID- 12560112 TI - Deferoxamine, allopurinol and oxypurinol are not neuroprotective after oxygen/glucose deprivation in an organotypic hippocampal model, lacking functional endothelial cells. AB - Reactive oxygen species-induced reperfusion injury of the brain is an important cause of neonatal morbidity and mortality following perinatal hypoxia-ischemia. Deferoxamine, allopurinol and oxypurinol have previously been shown to be neuroprotective in vivo during or directly after hypoxia-ischemia. To further characterize and more precisely elucidate whether the neuroprotective properties of these agents are mediated via neuronal and glial cells, or whether endothelial cells contribute to this effect, we tested their ability to protect CA1 neurons in organotypic hippocampal slices. Hippocampal slices obtained from 8-day-old rats were cultured for 7 days and exposed to oxygen/glucose deprivation for 50 min, or used as control slices. Cell damage was assessed at 48 h after oxygen/glucose deprivation using propidium iodide staining. At different time points following oxygen/glucose deprivation we administered dizocilpine, 6-cyano 7-nitroquinoxaline-2,3-dione, and alpha-phenyl-N-tert-butyl nitrone for validation purposes. Deferoxamine, allopurinol or oxypurinol were used as test substances. As expected, 89% and 98% protection was demonstrated with dizocilpine present during or during/after oxygen/glucose deprivation resp. alpha-Phenyl-N tert-butyl nitrone administered during/after oxygen/glucose deprivation provided 44% protection. However, iron chelation with deferoxamine and inhibition of xanthine oxidase by allopurinol or oxypurinol did not confer neuroprotection. The neuroprotective effect of deferoxamine, allopurinol or oxypurinol, as seen in vivo, may be obtained via inhibition of the production of damaging factors by blood born substances or endothelial cells. PMID- 12560113 TI - Cortical electrical stimulation alters erythrocyte perfusion pattern in the cerebral capillary network of the rat. AB - The effect of direct cortical electrical stimulation on the pattern of erythrocyte perfusion in the capillary network of the rat cerebral cortex was studied by fluorescence intravital video-microscopy. The movement of fluorescently labeled red blood cells (FRBCs) in individual capillaries 50-70 microm subsurface in the dorsal somatosensory cortex was visualized using a closed cranial window. Cortical stimulation electrodes were placed on opposite sides of the window. FRBC velocity (mm/s) and supply rate (cells/s) were measured in 51 capillaries from six rats before and during electrical stimulation of increasing intensities (15-s trains of 3-Hz, 3-ms, 0.5-5.0-mA, square pulses). FRBC velocity, supply rate, and the instantaneous capillary erythrocyte content (lineal cell density, LCD, cells/mm) increased with the stimulation current and reached maxima of 110, 160 and 33% above control, respectively. Capillaries with low resting velocity showed a greater response than those with high resting velocity. The fraction of capillaries in which FRBC velocity increased was not constant, but increased with the stimulation current, as did the magnitude of the velocity change in these capillaries. A few capillaries showed a negative FRBC velocity response at stimulations <4 mA. These results suggest that a robust rise in the fraction of responding (engaged) capillaries and a smaller rise in the capillary LCD contribute to neuronal activation-induced cortical hyperemia. Thus, capillary engagement and erythrocyte recruitment appear to represent important components of the cortical functional hyperemic response. These results provide insight into some of the specific hemodynamic changes associated with functional hyperemia occurring at the capillary level. PMID- 12560114 TI - Dependence on morphine leads to a prominent sharing among the different mechanisms of long-term potentiation in the CA1 region of rat hippocampus. AB - Here, we examined chronic exposure to morphine to determine if this treatment shifted LTP mechanism in the CA1 field in vitro. Long-term potentiation (LTP) of population spikes induced by a 200 Hz theta pattern primed bursts (PBs) stimulation. Verapamil was used to isolate NMDA-dependent LTP. In control slices, a 200 Hz tetanus induced a compound potentiation, consisted of two pharmacologically separable components: nmdaLTP and vdccLTP. LTP in slices taken from morphine dependent rats was completely abolished by either APV or verapamil. These data suggest that morphine dependence in rats does not interfere with the induction and maintenance of hippocampal CA1 LTP. While in control rats both NMDA and voltage-dependent Ca(2+) channel (VDCC) antagonists must have been used concurrently to prevent the induction of LTP, in morphine-dependent rats, each of the antagonist could prevent the LTP induction suggesting a tighter coupling between these two calcium influx regulating processes. PMID- 12560115 TI - Organization of the ectostriatum based on afferent connections in the zebra finch (Taeniopygia guttata). AB - In birds with laterally-located eyes, such as zebra finches and pigeons, the tectofugal visual pathway is the most prominent route from the retina to the telencephalon. However, little is known about exactly how the visual information is processed in this pathway, especially at the core region of the ectostriatum (Ec) in the telencephalon. In order to reveal a detailed organization of Ec, we decided to systematically analyze the afferent connections of Ec by injecting small amounts of sensitive tracers (biotinylated dextran amine and cholera toxin subunit B) selectively into different regions of Ec and the thalamic center of the tectofugal pathway (the nucleus rotundus, Rt). The present study revealed a clearer picture of the organization of Ec subdivisions than previously known. The present results showed that the anterior portion of Rt sent a heavy projection to the ventral region of the anterior Ec, whereas the more caudal subdivisions of Rt sent projections to more caudal and dorsal portions in Ec. The results suggest that Ec subdivisions appear to be arranged along an axis 'rotated' in the anterior direction, almost parallel to other major telencephalic laminae. These results may clarify the physiological and chemical heterogeneity of Ec found in the previous studies. The present findings also provide an insight into the possible organization of a visual processing center in a non-mammal. PMID- 12560116 TI - Central sympathetic chemosensitivity and Kir1 potassium channels in the cat. AB - The possible involvement of potassium channels in central chemosensitivity, with special reference to the Kir1.1 potassium channel, was investigated by studying the CO(2) response of presympathetic neurons in the rostroventrolateral medulla (RVLM) in the absence or presence of various K(+) channel inhibitors. Synaptic input to RVLM neurons was blocked by local injection of omega-agatoxin and omega conotoxin. Activity of RVLM neurons was measured by recording the electrical activity in preganglionic (WR-T(3)) or postganglionic (renal) sympathetic nerves after perfusion of the lower brainstem via the left vertebral artery with CO(2) enriched saline solution. Unspecific K(+) channel blockade by BaCl(2) reduced the excitatory response of sympathetic activity after CO(2)-perfusion to 56% of control. A quantitatively similar inhibition of the central CO(2) response was obtained after administration of 9-fluorenylmethylchloroformate (FMOC-Cl) which eliminates pH sensitivity of Kir1 and Kir4.1. Furthermore, two structurally different Kir1 inhibiting toxins, tertiapin and Lq2, also reduced the central CO(2) response to approximately 50% of control. In contrast, charybdotoxin (CTX) had no effect on the CO(2) response. Using RT-PCR the expression of mRNA homologous to rat Kir1 mRNA was identified in the cat medulla oblongata. These data suggest that a modulation of potassium channel activity possibly via Kir1 may contribute to central chemosensitivity. PMID- 12560117 TI - Vestibular nucleus projections to the Edinger-Westphal and anteromedian nuclei of rabbits. AB - The Edinger-Westphal nucleus (EW), anteromedian nucleus (AM) and adjacent neurons in the ventral tegmental area (VTA) are sources of preganglionic parasympathetic innervation of intraocular smooth muscle, including blood vessels, pupillary muscle and the ciliary body in mammals. They also have central connections that are believed to affect parasympathetic outflow indirectly. This study utilized anterograde transport of biotinylated dextran amine and Phaseolus vulgaris leucoagglutinin to demonstrate direct projections from the vestibular nuclei to the Edinger-Westphal and anteromedian nuclei in rabbits. The rabbit AM and adjacent VTA contain moderate to intensely choline acetyltransferase (ChAT) immunopositive neurons. The rabbit EW, by contrast, is nearly devoid of ChAT immunopositive neurons. Vestibular nucleus projections to these regions originate from all levels of the superior, medial and lateral vestibular nuclei, but do show topographic organization. The densest terminations were observed in AM and the ventral and central aspects of EW. The projections to AM terminate in both ipsilaterally and in a narrow paramedian region. Predominantly ipsilateral terminations were observed in VTA. Terminations on ChAT-positive cells in AM and VTA were verified in three rabbits. It is suggested that projections to some intensely ChAT-positive AM and VTA neurons may be a substrate for vestibular influences on lens accommodation, pupillary constriction and regulation of intraocular circulation during changes in posture and gravitoinertial challenges. The projections to ChAT-negative (and weakly immunoreactive) cells in AM, VTA and EW, on the other hand, are likely to contribute vestibular signals to a variety of motor responses via descending pathways. PMID- 12560118 TI - Tetrodotoxin-resistant sodium channels Na(v)1.8/SNS and Na(v)1.9/NaN in afferent neurons innervating urinary bladder in control and spinal cord injured rats. AB - Tetrodotoxin-resistant (TTX-R) sodium channels Na(v)1.8/SNS and Na(v)1.9/NaN are preferentially expressed in small diameter dorsal root ganglia (DRG) neurons. The urinary bladder is innervated by small afferent neurons from L6/S1 DRG, of which approximately 75% exhibit high-threshold action potentials that are mediated by TTX-R sodium channels. Following transection of the spinal cord at T8, the bladder becomes areflexic and then gradually hyper-reflexic, and there is an attenuation of the TTX-R sodium currents in bladder afferent neurons. In the present study, we demonstrate that Na(v)1.8 is expressed in both bladder and non bladder afferent neurons, while Na(v)1.9 is expressed in non-bladder afferent neurons but is rarely observed in bladder afferent neurons. In spinal cord transected rats 28-32 days following transection, there is a decreased expression of Na(v)1.8 sodium channels in bladder afferents, but no change in the expression of Na(v)1.8 in non-bladder afferent neurons. Both bladder and non-bladder afferent neurons exhibit limited increases in Na(v)1.9 expression following spinal cord transection. These results demonstrate that the expression of TTX-R channels in bladder afferent neurons changes after spinal cord transection, and these changes may contribute to the increased excitability of these neurons following spinal cord injury. PMID- 12560119 TI - NMDA receptor blockade in intact adult cortex increases trafficking of NR2A subunits into spines, postsynaptic densities, and axon terminals. AB - Past in vitro studies have used immunofluorescence to show increased clustering of the NR1 subunits of NMDA receptors (NMDAR) following NMDAR blockade, indicating that NMDARs self-regulate trafficking to and from spines. However, since a substantial portion of spinous NMDAR subunits can reside at sites removed from plasma membranes, whether or not these immunofluorescent clusters are synaptic remains to be shown. Also, the NR2A/B subunits undergo activity dependent switching at synapses, indicating that their subcellular distribution may be regulated differently from the NR1 subunits. We examined the issue of NMDAR autoregulation by determining whether in vivo NMDAR blockade enhances trafficking of the NR2A subunits toward spines and more specifically to postsynaptic densities (PSDs) of already mature synapses. Seven adult rats received unilateral intra-cortical infusion of the NMDAR antagonist, D-AP5 for 1/2-2 h and the inactive enantiomer or the solvent, alone, in the contralateral cortex. Using an electron microscope, approximately 5600 cortical spines originating from the two hemispheres of the seven adult animals were analyzed for the location of NR2A subunits. In six out of the seven cases analyzed, the D-AP5 treated neuropil exhibited increased immunolabeling at PSDs and a concomitantly great increase at non-synaptic sites within spines. NR2A subunits also increased presynaptically within 1/2 h but not after 1 h. These findings indicate that NR2A subunits in intact, adult cortical neurons are prompted to become trafficked into spines and axon terminals by NMDAR inactivity, yielding an increase of a readily available reserve pool and greater localization at both sides of synapses. PMID- 12560120 TI - Aggression-associated changes in murine olfactory tubercle bioamines. AB - The relationship between changes in regional brain bioamine levels and the expression of intraspecies aggressive behavior was evaluated in two murine models. In one study, normal male mice were maintained either in aggregate (i.e., normal, intraspecies social behavioral controls) or isolated (i.e., developed, non-social intraspecies aggressive 'fighter' behavior) housing environments, and the accompanying changes in both olfactory tubercle (OT) and hypothalamic (HYPOTH), norepinephrine (NE), dopamine (DA) and serotonin (5-HT) concentration indices quantitated by high-performance liquid chromatography (HPLC) for analysis of behavior-related alterations in localized bioamine deposition loci. Intact mice which had been housed in isolation cages and which exhibited aggressive, intraspecies reflexive-biting ('fighter') behavior when introduced to a novel (stimulus) animal, exhibited significant (P<0.05) elevations in NE levels, and depressed DA concentrations, in the OT regions relative to aggregated controls, indicating an intrinsic social influence on the maintenance of basal adrenergic indices at this neural locus. No changes in 5-HT levels were indicated between control and aggressive, isolated 'fighter' groups in either OT or HYPOTH loci. In addition, the NE and DA levels in the HYPOTH samples of both control and aggressive groups were found to be comparable. In the second study, utilizing an alternate type of aggression-induced murine model, changes in bioamine parameters were determined from samples obtained from aggregated, olfactory-bulbectomized (Obx) mice which are recognized to exhibit an overt, intraspecies, reflexive biting behavior as compared to sham-operated (control) mice housed under identical conditions. In these studies, Obx-mice exhibited a significant increase in 5-HT levels in the OT relative to sham-operated controls, but similar NE and DA concentrations. In addition, all hypothalamic bioamine indices were found to be comparable between control and Obx groups. These data, collected for both isolation-developed, and experimentally-induced (i.e., OBX), intraspecies aggressive models, indicate that the distinctive types of aggressive behaviors displayed by these two murine models are accompanied by specific alterations in regional bioamine levels within the OT of these groups, relative to controls. These data suggest that the specific type of overt aggressive behavior demonstrated by these models may be causally related to the identified changes in bioamine concentrations in the forebrain regions of the CNS, in loci recognized to participate in environmental recognition and social processing activities. PMID- 12560121 TI - Histochemical localization of neurons with zinc-permeable AMPA/kainate channels in rat brain slices. AB - Zinc modulates neurotransmission and may trigger neurodegeneration following brain injuries. Therefore, it is important to understand zinc dynamics in an anatomical context. Using a histochemical procedure on stimulated slices, we visualized zinc influx into neurons 'in situ'. Hippocampal, neocortical and cerebellar slices were loaded with zinc and stimulated with compounds known to open zinc-permeable channels. Zinc was then visualized by histochemical precipitation methods. Kainate stimulation labelled CA1 hippocampal pyramidal neurons, as well as subpopulations of interneurons in the hilus, CA1 and CA3 fields. Interneurons in the neocortex and many cell types of the cerebellum were also labelled. However, neither NMDA nor 50 mM K(+) stimulation resulted in comparable zinc accumulation in neurons. Immunofluorescent colocalization of parvalbumin with kainate-induced zinc staining in the hippocampus and neocortex showed a subset of zinc-sensitive neurons, positive for parvalbumin. These results confirm that zinc permeation through calcium-permeable AMPA/kainate channels constitutes a predominant route of zinc entry into different cell types. Furthermore, this technique provides a useful and versatile histochemical approach to assess the cell subpopulations of the central nervous system particularly sensitive to zinc accumulation under normal or pathological conditions. PMID- 12560123 TI - Influence of dopamine on ventrolateral thalamic inputs in cat motor cortex. AB - Neuronal activity in several brain regions is modulated by dopaminergic inputs. When single neuronal activity/20 trials of single-pulse ventrolateral thalamic (VL) stimulation was extracellularly recorded in the in vivo, anesthetized cat motor cortex, iontophoretic application of dopamine (DA) elicited either suppression or, in a fewer instances, facilitation of evoked unitary responses. The predominant inhibition exerted by DA appeared to be consistent for successive trials, and a D(1), D(2), and D(1)/D(2) receptor antagonist restored the effect, thereby reflecting a possible coexistence of two DA receptors. By contrast, only a fewer neurons' response to DA displayed facilitation, which was not attenuated by DA antagonists. Moreover, subsequent trials with receptor agonist and antagonists induced inconsistent effects. Except for the jitters, single unit spikes showed invariant latency, which was constant during all recording parameters, and the mean latency remained unchanged. The modulatory effects mediated by DA did not reveal any substantial difference between short- and long latency responses. Both pyramidal tract neurons and non-pyramidal tract neurons, determined on the basis of antidromic potentials from the pyramidal tract, responded to DA essentially in a similar manner. It appears that DA overall inhibits cat motor cortical neuronal activity in response to VL inputs. We propose that such DAergic inhibition of thalamocortical excitation in the motor cortex could be critical for ongoing sensorimotor transformation. PMID- 12560122 TI - Effects of chronic ethanol consumption on rat GABA(A) and strychnine-sensitive glycine receptors expressed by lateral/basolateral amygdala neurons. AB - It is well known that the anxiolytic potential of ethanol is maintained during chronic exposure. We have confirmed this using a light-dark box paradigm following chronic ethanol ingestion via a liquid diet. However, cessation from chronic ethanol exposure is known to cause severe withdrawal anxiety. These opposing effects on anxiety likely result from neuro-adaptations of neurotransmitter systems within the brain regions regulating anxiety. Recent work highlights the importance of amygdala ligand-gated chloride channels in the expression of anxiety. We have therefore examined the effects of chronic ethanol exposure on GABA(A) and strychnine-sensitive glycine receptors expressed by acutely isolated adult rat lateral/basolateral amygdala neurons. Chronic ethanol exposure increased the functional expression of GABA(A) receptors in acutely isolated basolateral amygdala neurons without altering strychnine-sensitive glycine receptors. Neither the acute ethanol nor benzodiazepine sensitivity of either receptor system was affected. We explored the likelihood that subunit composition might influence each receptor's response to chronic ethanol. Importantly, when expressed in a mammalian heterologous system, GABA(A) receptors composed of unique alpha subunits were differentially sensitive to acute ethanol. Likewise, the presence of the beta subunit appeared to influence the acute ethanol sensitivity of glycine receptors containing the alpha(2) subunit. Our results suggest that the facilitation of GABA(A) receptors during chronic ethanol exposure may help explain the maintenance of ethanol's anti-anxiety effects during chronic ethanol exposure. Furthermore, the subunit composition of GABA(A) and strychnine-sensitive glycine receptors may ultimately influence the response of each system to chronic ethanol exposure. PMID- 12560124 TI - Local inflammation increases vanilloid receptor 1 expression within distinct subgroups of DRG neurons. AB - Vanilloid receptor 1 (VR1) is essential to the development of inflammatory hyperalgesia. We investigated whether inflammation can increase in VR1 positive neuronal profiles in rat DRG neurons using histochemical methods. We also used size frequency analysis and double staining with several neuronal markers to investigate whether or not inflammation alters VR1 expression. Inflammation induced a 1.5-fold increase in percentage of VR1-like immunoreactivity (LI) positive profiles per total neuronal profiles, suggesting that the number of heat and pH sensitive neurons increase during inflammation. Area frequency histograms showed that VR1 expression increased in small and medium-sized neurons after inflammation. Double labeling of VR1 with NF200 showed that VR1 positive neurons with NF200 positive profiles significantly increased, indicating that the medium sized VR1 positive neurons were neurons with myelinated A-fibers. Local inflammation thus increases in VR1 protein level within distinct subgroups of DRG neurons that may participate in the development and maintenance of inflammatory hyperalgesia. PMID- 12560125 TI - Effects of exogenous creatine on population spike amplitude and on postanoxic hyperexcitability in brain slices. AB - In in vitro rat hippocampal slices, a short period of transient anoxia caused a lasting increase in the amplitude of the compound action potential (population spike, PS) that was evoked in CA1 by stimulation of the Schaffer collaterals. No such increase was seen over a comparable period of time in slices that were not subjected to anoxia. The appearance of such an increase was dependent on the duration of anoxia. Anoxia of 1 min duration did not cause any increase, anoxia lasting 2 min caused a nonsignificant increase, while 3 min of anoxia caused a lasting and statistically significant increase in PS amplitude. Addition of creatine, a compound that is known to afford protection against severe neuronal damage from longer periods of anoxia, prevented PS potentiation at a concentration of 10 mM, but not at a concentration of 1 mM. In addition, while 1 mM creatine by itself did not show any effect on PS amplitude of control slices, 10 mM creatine decreased PS amplitude also in such control slices, that had not been exposed to anoxia. These data demonstrate that this postanoxic hyperexcitability is caused by mechanisms that are little sensitive to the protection that in other contexts is provided by creatine. We suggest that understanding the mechanisms of postanoxic hyperexcitability may help understand the pathophysiology of the epileptic seizures that sometimes occur at the time of an ischemic stroke. PMID- 12560126 TI - Stereotaxic localization of corticosterone to the amygdala enhances hypothalamo pituitary-adrenal responses to behavioral stress. AB - The amygdala is involved in behavioral, autonomic, and neuroendocrine responses to stressful stimuli. The goal of the current study was to determine the effect of directly elevating glucocorticoids in the amygdala on hypothalamo-pituitary adrenocortical (HPA) responses to the elevated plus maze, a behavioral stressor known to activate the amygdala. Micropellets (30 microg) of crystalline corticosterone or cholesterol (control) were implanted bilaterally at the dorsal margin of the CeA in male Wistar rats; vascular catheters were also placed at this time. Five days post-surgery, blood samples were drawn at 07:00 and 19:00 h to assess diurnal rhythm of plasma corticosterone. At 7 days post-implantation, rats were subjected to behavioral stress using an elevated plus maze and blood was collected 15 min prior to stress, and at 15, 45, and 90 min after the initiation of the stressor. Corticotropin releasing factor (CRF) and arginine vasopressin (AVP) mRNA levels were analyzed by in situ hybridization in the medial parvocellular division of the hypothalamic paraventricular nucleus (mpPVN) in corticosterone- and cholesterol-implanted rats either not exposed to the elevated plus maze (control) or 4 h post-behavioral stress. Localization of corticosterone to the amygdala had no effect on diurnal rhythm of corticosterone secretion. Behavioral stress significantly increased peak plasma corticosterone levels in both groups to a similar level. However, in the corticosterone implanted rats, plasma corticosterone concentrations at 45 and 90 min post-stress were significantly greater compared to control rats indicating a prolonged corticosterone response to behavioral stress. In non-stressed rats, corticosterone delivery to the amygdala elevated basal CRF mRNA in the mpPVN to levels similar to those observed post-stress in control animals; no further increase was observed in CRF mRNA following stress. Behavioral stress resulted in a significant elevation in CRF mRNA in cholesterol controls. Basal AVP mRNA levels were unaffected by corticosterone implants. AVP mRNA did not increase in cholesterol implanted rats in response to behavioral stress. However, AVP mRNA levels were higher in corticosterone implanted rats post stress compared to cholesterol treated controls. In conclusion, direct administration of corticosterone to the amygdala increases plasma corticosterone in response to a behavioral stressor without altering the diurnal rhythm in plasma corticosterone. Elevated basal levels of mpPVN CRF mRNA, and the induction of a mpPVN AVP mRNA response to the behavioral stressor implicate enhanced ACTH secretagogue expression in the increased HPA response to corticosterone modulation of amygdala function. PMID- 12560127 TI - Neuroprotective effects of a new synthetic peptide, CMX-9236, in in vitro and in vivo models of cerebral ischemia. AB - NGF (nerve growth factor) and BDNF (brain-derived neurotrophic factor) are protein molecules (MW 26 and 13.6 kDa, respectively) that are neuroprotective in the middle cerebral artery occlusion (MCAO) rat stroke model. Their mechanism of action involves the activation of transcription factor AP-1 that turns on neuronal growth genes. In our ongoing studies we are designing short peptides that mimic some of the properties of full-length neurotrophic factors. We have synthesized a neuroprotective 14-amino acid peptide (CMX-9236) with an N-terminal docosahexaenoic acid (DHA). DHA enhances entry through the blood-brain barrier. Using primary rat brain cortical cultures and a fluorescent assay we found that CMX-9236 can counteract the excitotoxic effects of glutamate or kainate, reversing the intracellular accumulation of Ca(2+) to normal levels. Administration (i.v.) of CMX-9236 post initiation of ischemia reduced the lesion volumes from 178+/-50 to 117+/-55 mm(3) in the temporary rat MCAO model (90 min), and from 216+/-58 to 127+/-57 mm(3) in the permanent (24 h) model for stroke, corresponding to 34+/-28% (P=0.01) and 41+/-19% (P=0.038) reductions of the infarct volumes. Neurological behavior scores showed 57 and 47% improvements for treated temporary and permanent models, respectively. Dose-response studies indicated a 60-fold activation of AP-1 transcription factor in cells treated with 100 ng/ml of the peptide. These studies illustrate that a small peptide can function as a neuroprotective agent and an activator of a beneficial signal transduction pathway. PMID- 12560128 TI - Possible involvement of nitric oxide in the central salt-loading-induced cardiovascular responses in conscious rats. AB - The objective of this study was to elucidate the possible involvement of nitric oxide (NO) in the cardiovascular responses induced by central salt loading. Direct perfusion of the hypothalamic paraventricular nucleus (PVN) region with hypertonic saline (0.3 or 0.45 M) was performed in conscious rats by using an in vivo brain microdialysis technique. The extracellular concentration of NO metabolites in the PVN region was measured, as were the blood pressure (BP) and heart rate (HR). Perfusion of 0.45 M saline increased the BP, HR, and NO metabolite levels in the PVN region; however, perfusion of 0.3 M saline enhanced only the level of NO metabolites but did not induce changes in the BP and HR. Next, we determined whether the NO was involved in the cardiovascular responses induced by hypertonic saline. Pretreatment with N(G)-methyl-L-arginine (L-NMMA), an inhibitor of NO synthase, attenuated the increases in the BP and HR induced by direct perfusion of 0.45 M saline, while direct infusion of 3 morpholinosyndnonimine (SIN-1, a NO donor) in the PVN region induced increases in the BP and HR. These results suggest that local perfusion of the PVN region with hypertonic saline elicits a local release of NO, which may be carried out by activating nitric oxide synthase to produce cardiovascular responses. PMID- 12560129 TI - Involvement of NMDA-nitric oxide pathways in the development of tactile hypersensitivity evoked by the loose-ligation of inferior alveolar nerves in rats. AB - To investigate whether or not NMDA/nitric oxide (NO) pathways in the trigeminal system are involved in the development and/or maintenance of such pathological pain states as the hyperalgesia and allodynia observed after dental surgery, loose-ligation on the left inferior alveolar nerves of rats were performed. The responses to mechanical stimulation were then measured using von Frey filaments. Hypersensitivity to tactile stimulation developed on the ipsilateral side in ligated animals 5 days after surgery and lasted for at least 30 days. In addition, the effects of drugs on these pain states during the period 2-3 weeks following surgery were investigated. As a result, it was observed that tactile hypersensitivity was inhibited by the intraperitoneal (i.p.) administration of both MK-801 hydrogen maleate (0.05-0.1 mg/kg) and N(G)-monomethyl-L-arginine acetate (L-NMMA: 10-100 mg/kg). Still further, NO production and the number of neuronal NO synthase (nNOS)-positive neurons in the trigeminal nucleus caudalis (SpVc) was evaluated. As a result of these experiments, it was found that the changes in NO levels evoked by the intravenous infusions of N-methyl-D-aspartate (NMDA; 10 mg/kg) and MK-801 (0.5 mg/kg) were significantly larger in the loose ligated rats compared to the sham-operated rats. Moreover, the number of nNOS positive neurons was found to have increased on the ipsilateral side in layers I/II of the SpVc. These results would suggest that tactile hypersensitivity develops after inferior alveolar nerve injury and that NMDA receptor/NOS/NO production pathways in the SpVc may be involved in the development of such pathophysiological states. PMID- 12560130 TI - Serotonin depletion in rat hippocampus attenuates L-NAME-induced spatial learning deficits. AB - Inhibition of nitric oxide (NO) synthesis has been found to produce learning deficits in spatial tasks. Recent studies also suggest a regulatory effect of endogenous NO on hippocampal serotonin (5-HT) release and have shown that NO synthase (NOS) inhibitors increased extracellular levels of serotonin (5-HT) in the rat hippocampus. To clarify possible interactions between NO and 5-HT in the hippocampus on learning processes, the effect of selective hippocampal 5-HT depletion on NOS inhibition-induced spatial learning deficits was investigated. Rats received bilateral injections of 5,7-dihydroxytryptamine (5,7-DHT), a 5-HT neurotoxin, or its vehicle in the CA1 region of hippocampus following pretreatment with desipramine. Rats were subjected to 5 days of training in the Morris water maze (MWM); 4 days with the invisible platform to test spatial learning and the 5th day with the visible platform to test motivation and sensorimotor coordination. Nomega-nitro-L-arginine methyl ester (L-NAME), a NOS inhibitor, was administered to either sham-operated or 5,7-DHT-lesioned groups 30 min before training each day. Results showed that L-NAME significantly impaired the ability of rats to locate the hidden platform. This impairment was reversed by co-administration of mole equivalent dose of L-arginine, the NO precursor. Although the 5,7-DHT-induced lesion had no effect by itself on rat performance in the MWM, it attenuated the memory impairment caused by L-NAME. The observed effect suggests an interaction between NO and 5-HT in the hippocampus on spatial memory formation; however, the mechanism of interaction is still unclear and requires further investigation. PMID- 12560131 TI - Ethanol-induced impairment of spatial memory and brain matrix metalloproteinases. AB - The formation of spatial memory appears to be dependent upon an intact hippocampus capable of the specific biochemical changes associated with synaptic remodeling. Hippocampal damage results in the disruption of synaptic remodeling and the acquisition of spatial memory tasks. Ethanol also disrupts normal hippocampal functioning and spatial memory. The present investigation established a dose-response relationship between ethanol treatment and impairment of spatial memory as measured using the circular water maze task. Intraperitoneal ethanol doses of 1.5 and 2 g/kg significantly increased the latency and distance swam to find the submerged pedestal as compared with a 1 g/kg dose, and 0.15 M NaCl vehicle control treatments. On days 2, 4, and 6 of acquisition animals were sacrificed and brain tissues were retained from the hippocampus, prefrontal neocortex, and cerebellum for measurement of matrix metalloproteinases (MMPs). The results indicated that ethanol treatment interfered with MMP-9, but not MMP 2, activity in the hippocampus, and to a lesser degree in the prefrontal cortex. No changes in the cerebellum were measured. Elevations in MMP activity appear to be a prerequisite to reconfiguration of extracellular matrix cell adhesion molecules thought to be important in the process of synaptic plasticity, which in turn appears to be necessary for memory consolidation. Thus, ethanol-induced impairment in the acquisition of spatial memory tasks may, in part, be due to disruption of brain MMP activity. PMID- 12560132 TI - Ultrastructural features of synapse from dorsal parvocellular reticular formation neurons to hypoglossal motoneurons of the rat. AB - The dorsal parvocellular reticular formation (PCRt) receives projection of the trigeminal mesencephalic nucleus neurons. It contains the dorsal group of interneurons that integrate and coordinate activity of the oral motor nuclei. Ultrastructural features of synaptic connection from the dorsal PCRt neurons to the motoneurons of the hypoglossal nucleus (XII) were examined at both the light and electron microscopic levels in rats. Biotinylated dextran amine (BDA) was initially iontophoresed into the dorsal part of PCRt unilaterally. Seven days later horseradish peroxidase (HRP) was injected into the body of the tongue. After histochemical reaction for visualization of HRP and BDA, the BDA-labeled fibers and terminals were seen distributing bilaterally in XII with ipsilateral predominance. BDA-labeled terminals were closely apposed upon HRP retrogradely labeled somata and dendrites of the XII motoneurons. A total of 1408 BDA-labeled boutons were examined ultrastructurally, which had mean size of 1.22+/-0.37 microm in diameter. Five hundred-ninety three of these boutons in both the ipsilateral (n=401) and contralateral (n=192) XII were seen to synapse on both the dendrites and somata of HRP-labeled motoneurons. The vast majorities of synapses were axodendritic (98%, 580/593), while 2% of them were axosomatic. Of the 1408 BDA-labeled boutons, 69.6% of them were S-type boutons containing small clear and spherical synaptic vesicles and 30.4% of them were PF-type boutons containing pleomorphic and flattened synaptic vesicles. Approximately 64% of synapses between BDA-labeled boutons and HRP-labeled motoneurons were asymmetric, and 33% of synapses were symmetric. No axoaxodendritic or axoaxosomatic synaptic triad was observed. The present study illustrated the anatomical pathway and synaptological characteristics of neuronal connection between the dorsal PCRt premotor neurons and the XII motoneurons. Its functional significance in coordinating activity of XII motoneurons during oral motor behaviors has been discussed. PMID- 12560133 TI - Opposite metabolic changes in the habenula and ventral tegmental area of a genetic model of helpless behavior. AB - Congenitally helpless rats have been selectively bred to display an immediate helpless response to stress in order to model hereditary brain differences that contribute to depression vulnerability. Differences in regional brain metabolism between congenitally helpless and non-helpless rats were investigated using quantitative cytochrome oxidase histochemistry. The results indicated that congenitally helpless rats had 64-71% elevated metabolism in the habenula and a 25% elevation in the related interpeduncular nucleus. In contrast, helpless rats had 28% reduced metabolism in the ventral tegmental area (VTA) and 14-16% reductions in the basal ganglia and basolateral and central amygdala. The opposite metabolic changes in the habenula and ventral tegmental area may be especially important for determining the congenitally helpless rat's global pattern of brain activity, which resembles the metabolic activity pattern produced by dopamine antagonism. PMID- 12560134 TI - Rhizoma acori graminei and its active principles protect PC-12 cells from the toxic effect of amyloid-beta peptide. AB - The effects of water extracts of six medicinal herbs (Radix polygalae tenuifoliae, Radix salviae miltiorrhizae, Rhizoma acori graminei, Rhizoma pinelliae ternatae, Tuber curcumae and Scletrotium poriae cocos) on the cytotoxic action of Abeta(1-40) were tested with PC-12 cells. Only the extract of R. acori graminei (RAG) significantly decreased Abeta(1-40)-induced cell death. Further, eugenol and beta-asarone were isolated and identified as the major active principles. Both purified eugenol and beta-asarone protected PC-12 cells from the toxic effect of Abeta(1-40). Eugenol was active between 1 and 100 microM, and 10 microM eugenol gave approximately a 50% response. beta-Asarone was less potent and exhibited little, if any, activity at this concentration. Both eugenol and beta-asarone inhibited Ca(2+) intake by PC-12 cells: beta-asarone mainly inhibited basal Ca(2+) intake, whereas eugenol inhibited Abeta-induced Ca(2+) intake preferentially. These results suggest that eugenol may act by blocking Abeta-induced-Ca(2+) intake and provide a strong case for further pursuit of the therapeutic and prophylactic potentials of RAG and its active principles for the management of Alzheimer's disease. PMID- 12560135 TI - Cholecystokinin combined with serotonin in the hypothalamus limits accumbens dopamine release while increasing acetylcholine: a possible satiation mechanism. AB - Serotonin (5-HT) or cholecystokinin (CCK) injected in the hypothalamic paraventricular nucleus (PVN) inhibits feeding, but the mechanism is unknown. Prior research suggests that dopamine (DA) input to the nucleus accumbens (NAc) motivates behavior, and a component of that motivation circuit includes hypothalamic feeding systems. Acetylcholine (ACh) in the NAc, on the other hand, may act in part to inhibit feeding and generate satiety. If so, 5-HT and/or CCK in the PVN should lower extracellular DA or release ACh in the NAc. Rats were prepared with microdialysis probes in the NAc and injectors in the PVN. Serotonin (7.75 microg) or CCK-8 (0.12 microg) injected in the PVN significantly decreased ipsilateral accumbens DA (63 and 73% of baseline, respectively, without effect on ACh). However, 5-HT plus CCK injected in combination decreased DA to 72% (P<0.001) and simultaneously increased extracellular ACh to 128% of baseline (P<0.001). In later tests with the same doses and the same animals, unilateral PVN injections of 5-HT, CCK, or both combined, significantly inhibited food intake in the early dark period. The results suggest that 5-HT in the PVN acts as a neural modulator that primes a hypothalamic satiation system to respond to CCK when the gastrointestinal tract contains food to be digested. The synergistic action of 5-HT plus phasic CCK may then activate a circuit that simultaneously limits DA and releases ACh in the accumbens as part of the satiation process. PMID- 12560136 TI - Rat retinal ganglion cell loss caused by kainate, NMDA and ischemia correlates with a reduction in mRNA and protein of Thy-1 and neurofilament light. AB - Quantification of retinal ganglion cell (RGC) loss/survival following a defined insult to the retina is a prerequisite in order to allow a comparison to be made between the effectiveness of potential neuroprotective drugs. The purpose of the present study was to extend the characterisation of our previously published semiquantitative RT-PCR assay to assess RGC loss/survival. Comparisons were made between the total mRNA levels of the ganglion cell-specific markers Thy-1 and neurofilament light (NF-L) in the retina at specific times after an intravitreal injection of N-methyl-D-aspartate (NMDA) or kainate or after 45 min of ischemia/reperfusion and also between the levels of NF-L mRNA and protein at various times after NMDA injection. Changes in Thy-1 and NF-L immunoreactivities were also observed. NMDA, kainate and ischemia/reperfusion all caused a reduction in the retinal content of Thy-1 and NF-L mRNAs and immunoreactivities. An excellent correlation was observed between the levels of the two mRNAs after these treatments. After NMDA, loss of NF-L mRNA was shown to precede loss of NF-L protein but total loss of each marker was similar after 7 days. The results of the study demonstrate that injury and subsequent death of RGCs, which occurs after ischemia/reperfusion and after intraocular injection of NMDA or kainate, can be followed by measurement of total retinal levels of Thy-1 and NF-L mRNAs and NF-L protein. The assays provides accurate, practical and complementary methods for assessing the potential benefits of neuroprotective drugs on RGCs which have been injured by a variety of experimental modalities. PMID- 12560137 TI - PB-2, a polysaccharide fraction from lichen Flavoparmelia baltimorensis, peripherally promotes the induction of long-term potentiation in the rat dentate gyrus in vivo. AB - We have previously found that oral or intravenous administration of PC-2, a polysaccharide fraction purified from extracts of lichen Flavoparmelia caperata, facilitates the induction of long-term potentiation (LTP) in the dentate gyrus of anesthetized rats. PC-2 could be useful in the development of therapeutic drugs for senile dementia. However, it has been very difficult to obtain the material Flavoparmelia caperata because of its scarcity. In the present study, we therefore investigated whether PB-2, a polysaccharide fraction from another lichen Flavoparmelia baltimorensis, has similar biological effects. Oral administration of PB-2 (100-200 mg/kg) did not affect basal evoked potentials, but significantly promoted the induction of LTP following tetanic stimulation (30 pulses at 60 Hz) in the dentate gyrus of anesthetized rats. Intravenous injection of PB-2 (1-5 mg/kg) was also effective in promoting the induction of LTP, but intracerebroventricular injection of PB-2 (1-2 mg/brain) was ineffective. PB-2, as well as PC-2, should be valuable in identifying factors that promote synaptic plasticity in the hippocampus. PMID- 12560138 TI - Serotonergic signaling inhibits hyperalgesia induced by spinal cord damage. AB - Although dysesthesia is one of the most serious problems in patients with spinal cord injury, most of them being unresponsive to conventional treatments. In this study, we established a rat thoracic spinal cord mild-compression model that revealed thermal hyperalgesia in the hind limb. The thoracic spinal cord was compressed gently, using a 20 g weight for 20 min. The withdrawal latency of the thermal stimulation of the bilateral hind-limb was monitored using Hargreaves' Plantar test apparatus. In this model, thermal-hyperalgesia was observed for 1 week after the injury. The spinal cord injury-induced thermal-hyperalgesia was mimicked by the intrathecal application of metergoline, a non-selective 5-HT antagonist, 1-(2-methoxyphenyl)-4-[4-(2-phthalimido) butyl]-piperazine hydrobromide (NAN190), a selective 5-HT1 antagonist, and 3-tropanyl-3,5 dichlorobenzoate (MDL72222), a selective 5-HT3 antagonist. Intraperitoneal application of fluvoxamine maleate, a selective serotonin reuptake inhibitor, reduced the intensity of hyperalgesia induced by spinal cord injury. The inhibitory effect of fluvoxamine maleate on thermal hyperalgesia was prevented by the application of the aforementioned nonselective or selective 5-HT receptor antagonists. Intrathecal application of fluvoxamine maleate and selective 5-HT receptor agonists, i.e., 8-hydroxy-2-(di-n-proplyamino)-tetralin hydrobromide (8 OH-DPAT: 5HT-1 selective) and 2-methyl-5-hydroxytryptamine maleate (2-m-5-HT: 5HT 3 selective), inhibited the spinal cord injury-induced hyperalgesia. These results suggest that the change in the descending serotonergic signal plays an important role in hyperalgesia after the spinal cord injury, and that the application of selective serotonin reuptake inhibitors will be one of the candidates for new therapeutic methods against post-spinal cord injury dysesthesia. PMID- 12560139 TI - Rat Schwann cells in bioresorbable nerve guides to promote and accelerate axonal regeneration. AB - A micro-structured, biodegradable, semipermeable hollow nerve guide implant was developed to bridge nerve lesions. Quantitative comparison of cell migration and axonal growth using time lapse video recording in vitro revealed that axons grow eight times faster than neuritotrophic Schwann cells migrate. To accelerate regeneration, purified Schwann cells are best injected into nerve guides before implantation. Nerve guides made from resorbable poly-lactide-co-glycolide support Schwann cell attachment, cell survival, and axonal outgrowth in vitro. The therapeutic concept aims at the development of an 'intelligent neuroprosthesis' that first mediates regeneration and then disappears. PMID- 12560140 TI - Effects of minocycline alone and in combination with mild hypothermia in embolic stroke. AB - Inflammatory reactions occurring in the brain after ischemia may contribute to secondary damage. In the present study effects of minocycline, an anti inflammatory agent, alone or in combination with mild hypothermia, on focal embolic brain ischemia have been examined. Focal ischemic injury was induced by embolizing a preformed clot into the middle cerebral artery (MCA). Infarct volume was measured at 48 h after the injury. Administration of minocycline alone or minocycline plus mild hypothermia reduced infarct volume significantly. However, mild hypothermia in combination with minocycline did not show any additive effect. These results suggest that minocycline is beneficial in focal ischemic brain injury, and the lack of the enhanced neuroprotection may be due to the brief exposure to hypothermia. PMID- 12560141 TI - Success and complications of four-duct ligation for sialorrhea. AB - OBJECTIVE: Sialorrhea can have a significant negative effect on quality of life, impeding social interactions and severely limiting employment opportunities. Many surgical options to control sialorrhea have been reported. One of the newest procedures is combined ligation of the submandibular and parotid ducts, reported only once in the literature in one series of five patients to date. We have compared results in our first 21 patients undergoing this procedure with results reported in the literature for other procedures to treat sialorrhea. MATERIALS AND METHODS: We retrospectively reviewed medical records of all 21 children we treated with four-duct ligation, a relatively simple intraoral procedure to control sialorrhea, between August 1999 and September 2000 and contacted primary caregivers by telephone to answer a questionnaire regarding objective and subjective results of surgery. Surgery was considered successful when caregivers rated patients as 'much better' or 'better' after surgery. RESULTS: Follow-up was completed in all 21 of the patients 1-14 months after surgery. The success rate of four-duct ligation ('much better' or 'better' after surgery) was 81%, and no patient's sialorrhea problem was worse after surgery. Major complications occurred in two (10%) of the patients (one ranula and one case of sialoadenitis), which were both successfully treated surgically. Minor complications occurred in four (19%) of the patients, tongue swelling that prolonged hospitalization, a ranula that resolved, and prolonged submandibular gland swelling that resolved (two cases). More than half of patients were discharged the day of or the day after surgery. CONCLUSIONS: Four-duct ligation should be considered when surgery is indicated to treat sialorrhea. PMID- 12560142 TI - The changing indications for paediatric tracheostomy. AB - OBJECTIVE: To investigate whether the incidence and indications for paediatric tracheostomy in this unit have changed over recent years. METHODS: All paediatric tracheostomies performed between 1993 and 2001 were identified from our departmental database. The indications for these were ascertained by retrospective case note review. RESULTS: Over the 9-year period studied 362 tracheostomies were performed, the number increased slightly between the first and second half of the period, with peaks in 1997 and 1999. The commonest indication was prolonged ventilation due to neuromuscular or respiratory problems. CONCLUSIONS: This large series shows that the increase in frequency of paediatric tracheostomy performed in this unit over the past decade has been due to conditions such as subglottic and tracheal stenosis, respiratory papillomatosis, caustic alkali ingestion and craniofacial syndromes. Conditions in which tracheostomy are now less common are subglottic haemangioma and laryngeal clefts. Prolonged ventilation remains the commonest indication overall. PMID- 12560143 TI - The effect of aryepiglottoplasty for laryngomalacia on gastro-oesophageal reflux. AB - OBJECTIVE: to investigate whether relief of airway obstruction in laryngomalacia by aryepiglottoplasty affects gastro-oesophageal reflux. METHODS: a prospective study of consecutive infants and children with suspected laryngomalacia. Gastro oesophageal reflux was measured before and after diagnostic microlaryngobronchoscopy and aryepiglottoplasty. RESULTS: of the six cases who underwent aryepiglottoplasty and completed the study, three had significant pre operative reflux according to age. In this group the reflux improved significantly after aryepiglottoplasty. In the other three cases, reflux was not age-significant pre-operatively nor did it change significantly post-operatively. CONCLUSIONS: when partial airway obstruction due to laryngomalacia co-exists with gastro-oesophageal reflux, treatment of the airway problem improves respiratory symptoms in all cases and reduces gastro-oesophageal reflux in patients with age significant reflux. This suggests that there are two clinical groups, those with severe, age-significant reflux, possibly caused by airway obstruction, whose gastro-oesophageal reflux benefits from aryepiglottoplasty; and those whose reflux is physiological and not influenced by aryepiglottoplasty. Therefore aryepiglottoplasty can be expected to reduce gastro-oesophageal reflux in those infants with laryngomalacia who have age-significant reflux. PMID- 12560144 TI - Identification of hearing loss in newborns by transient otoacoustic emissions. AB - OBJECTIVE: More than 90% of congenital hearing loss cases are of cochlear origin. There are two methods for newborn hearing screening: the transient otoacoustic emission (TEOAE) or (ABR) screening. When TEOAE is used for hearing screening patients, newborn with a neural hearing loss are not discovered. MATERIALS: In the present study TEOAEs were obtained from 3,048 newborns from both ears in patients with and without risk factors for hearing loss in the history. All newborns who did not pass TEOAE in the 2nd screening (uni or bilaterally) underwent additional audiologic tests. RESULTS: In the first screening 150 (4.5%) of the newborns newborn did not pass the screening and 30 (0.98%) did not pass in the second screening. In nine newborns with unilateral absent TEOAE and in two newborns a deafness was confirmed, with one side cochlear and on the other side retrocochlear. Both patients received cochlear implants before the 2nd year of age. In 21 newborns with absent TEOAE bilaterally, five had moderate sensorineural hearing loss bilaterally and in nine patients profound hearing loss (90-100 dB) or deafness was confirmed. Of 3,048 newborns there were 1,355 with a risk of hearing loss and in 12 (0.88%) newborns bilateral hearing loss or deafness was confirmed; of the 1663 newborns without risk of hearing loss in the history, in four (0.24%) newborn deafness or bilateral sensorineural hearing loss in the range of 45-65 dB were confirmed. CONCLUSION: When the newborn has an absent TEOAE uni or bilaterally, we need to inform the parents and to recommend additional screening and other audiologic tests to confirm or exclude hearing loss. PMID- 12560145 TI - Risk factors for thyroglossal duct remnants after Sistrunk procedure in a pediatric population. AB - To evaluate recurrence after surgery for thyroglossal duct cyst (TDC) we performed a retrospective chart review. Seventy four patients between 0.5 and 8.5 years of age presenting with a midline neck cyst underwent a Sistrunk procedure for a preoperative diagnosis of TDC. Fifty-seven had histologically confirmed TDC (mean age of the population: 4+/-1.5 years, mean follow-up: 6 years and 8 months). Recurrence occurred in 15% of the cases of histologically confirmed TDC. Four individual risk factors have been identified: number of infection before surgery [more than 2 episodes (P<0.05)]; preliminary surgical procedure (P<0.05); age [less than 2 years (P<0.05)] and multicystic lesion on histopathology (P<0.01). The two first factors being correlated, the risk of relapse might be lowered by a wide excision performed before any infection in children over 2 years. PMID- 12560146 TI - Mandibular fractures in children: long term results. AB - Mandibular fractures in children treated in our department between March 1994 and January 2001 were retrospectively studied. Age, sex, type of fracture, etiology and evolution after treatment, functional mobility and maximal mouth opening were recorded. The population consisted of 19 patients who sustained 30 fractures. The patients ages ranged from 1.5 to 18 years. The mean time of follow up was 28 months. The male to female ratio was 1.7:1. Traffic and bicycle accidents were the main causes of the fractures. The condyle was involved in 16% of the cases, the subcondylar region in 28%. Fractures were multiple in half of the cases. Isolated fractures of the condyloid joint were treated conservatively. For isolated subcondylar fractures, maxillomandibular fixation was the treatment in 40% of the cases. Otherwise, conservative functional treatment was used. Children with a combination of body and condyle fractures were treated by open reduction and maxillomandibular fixation. Neither infection nor retarded facial growth was observed. Only one case of ankylosis of the temporomandibular joint (TMJ) and one case of temporomandibular pain syndrome were recorded. Associated lesions might concern the extremities, the brain and the cervical spine. PMID- 12560147 TI - Acute mastoiditis caused by Moraxella catarrhalis. AB - Acute mastoiditis is the most frequent intratemporal complication of otitis media. The bacteriology of acute otitis media is changing continuously and it differs markedly from the bacteriology of acute mastoiditis. Moraxella catarrhalis (M. catarrhalis) is the third most common bacteria found in acute otitis media, and in recent years its importance as an etiological factor of acute otitis media has markedly increased in certain geographic areas. However, there are no reports of acute mastoiditis caused solely by M. catarrhalis. This report describes a case of a 2-year-old girl with acute mastoiditis and M. catarrhalis in the bacterial culture of middle ear effusion. PMID- 12560148 TI - Using a children's book to prepare children and parents for elective ENT surgery: results of a randomized clinical trial. AB - OBJECTIVE: We evaluated the effects of surgery preparation using a children's book on pre- and postoperative anxiety and distress in 2-10 years old children undergoing tonsillectomy and/or adenoidectomy and their mothers. METHODS: Parents of the experimental group were given the preparation book during the preoperative visit at the hospital, whilst control subjects did not receive the book. Data collection was conducted on the evening prior to surgery (T1), and the evening post surgery (T2). At these two points in time, mothers completed a self-designed feeling states checklist and the state anxiety scale of the State and Trait Anxiety Inventory to assess their perceptions of the child's distress and their own level of anxiety. At T2, nurses were asked to give ratings of patient caregiver variables, such as level of anxiety, cooperation with care or level of information. The sample under study consisted of 160 mother/child dyads in the experimental group and 240 controls. RESULTS: We found that mothers who received the book exhibited less self-reported state anxiety prior to the operation compared to mothers who did not. Simultaneously, children of the experimental group showed less distress in 4 of 11 feeling states. Nurses assessed the mothers of the experimental group to participate more in the child's care than control mothers. CONCLUSIONS: The results demonstrated that our preparation book can provide educational and anxiety-reducing benefits. Given the relatively low production costs and its easy administration, it can be recommended as a popular, practical and cost-efficient tool to prepare children and parents for surgery and hospitalization. PMID- 12560149 TI - Etiology of acute otitis media in childhood and evaluation of two different protocols of antibiotic therapy: 10 days cefaclor vs. 3 days azitromycin. AB - BACKGROUND: Acute otitis media (AOM) is a common childhood infection that is frequently treated by antibiotics. There are no prospective and comprehensive trials evaluating childhood AOM for etiologic pathogens and resistance pattern in Turkey. The aims of the study were to determine the bacterial etiologies and resistance patterns, and identify the efficacy and the relapse rates of 3 days of azitromycin and 10 days of cefaclor therapy in AOM. METHODS: This prospective, randomized, single-blind, open study was carried out in 78 cases of AOM. Mean age was 30.7+/-27 months. Tympanocentesis and aspiration of middle ear fluid (MEF) were used to obtain purulent material from the middle ear. Group 1 consisted of the cases (n=41) on azitromycin therapy and Group 2 (n=37) on cefaclor. Dosage of azitromycin was 10 mg/kg per day for 3 days and cefaclor 40 mg/kg per day for 10 days. The patients were evaluated on days 3-5 (second visit), day 10 (third visit), and day 30 (fourth visit) during follow-up. RESULTS: A total of 50 species were isolated from 44 of 78 cases from which materials were obtained (44/78; 56.4%). Most frequently isolated microorganism was Streptococcus pneumoniae (n=18; 36%), followed by Haemophilus influenzae (n=11; 22%), S. aureus (n=9; 18%), Moraxella catarrhalis (n=4; 8%), and group A beta-hemolytic streptococcus (GAS, n=4; 8%). Enterococcus faecalis was isolated from three cases and H. parainfluenzae from one. Penicillin and amoxicillin resistances of bacteria were found to be 40 and 36%, respectively. The frequency of penicillin and amoxicillin resistance in or = 0.5 microg/l. Additionally, plasma vitellogenin concentrations were significantly depressed in frogs exposed to 500 microg/l MXC. These data indicate that reproductive and endocrine dysfunction can occur in adult amphibia exposed to high concentrations of an environmental toxin with endocrine disrupting activity. Such effects may be indicative of the potential for adverse effects on amphibian wildlife exposed to environmental EDs. PMID- 12560168 TI - Acetylcholinesterase activity in copepods (Tigriopus brevicornis) from the Vilaine River estuary, France, as a biomarker of neurotoxic contaminants. AB - From April 1997 to June 1998, 14 measurements of acetylcholinesterase (AChE) enzymatic activity were performed with the copepod, Tigriopus brevicornis, collected at five stations in the Vilaine River estuary (South Brittany, France). Simultaneously, four chemical analyses of triazines and one analysis of total pesticides in water were undertaken. AChE activity levels in T. brevicornis were compared to the levels measured at a reference site not exposed to effluents from Vilaine River. Results reveal significant differences between AChE activity levels depending on location of stations in the plume of the river with an increasing gradient of activity from the upstream to the downstream stations, thus indicating that neurotoxic contaminants are mainly brought by the river. The average degree of AChE inhibition between the reference site and the most upstream site is 70-80% during spring in 1997 and 1998. In May 1997, live copepods from the different sites were brought back and transferred to clean seawater. After 14 days, recovery of AChE activity was almost total when compared to the control. Moreover, using a linear regression model and the atrazine concentration as marker of the presence of pesticides, low levels of AChE activity were significantly explained by atrazine concentration in water. PMID- 12560169 TI - Inter-population differences in Cd, Cr, Se, and Zn accumulation by the green mussel Perna viridis acclimated at different salinities. AB - Trace metal (Cd, Cr, Se, and Zn) uptake from the dissolved phase, assimilation efficiency from the dietary phase, and metal body burden, as well as the clearance rate, apparent water permeability and gill surface area were measured in the green mussel Perna viridis collected from two contrasting salinity sites in Hong Kong coastal waters with following acclimation in the laboratory at different salinities. The concentrations of metals were 1.2-6.4x greater in mussels collected from the low salinity site, i.e. Tai O, as compared with Tap Mun (high salinity) mussels. Influx of Cd and Zn from the dissolved phase increased with decreased salinity. Furthermore, Cr(VI) and Se (selenite) influx were also affected by decreased salinity, in particular below 17 psu, suggesting that speciation was not the only factor to effect metal uptake. Long-term acclimation to different salinity also had an effect on metal uptake. Mussels collected from Tap Mun (high salinity) accumulated metals 1.2-2.2x faster than mussels from Tai O (low salinity) at intermediate and high experimental salinities >17 psu. Metal assimilation efficiencies were unaffected by salinity variation, but gut passage times were significantly longer at low (10 psu) salinities, indicating some effect of lowered salinity on mussels gut physiology. Gill surface area, which was similar in both populations of mussels, and filtration rate, which was generally similar or higher in those groups with low metal uptake, could not explain the inter-population difference in metal accumulation. Although there was no significant difference, due to high inter individual variability, the apparent water permeability of the Tap Mun population was on average about 1.6x greater compared with Tai O mussels, and may partially account for the difference in metal uptake between these two populations acclimated at salinity >17 psu. Thus, the effect of salinity on metal uptake is dependent on metal biogeochemistry as well as a range of physiological responses. There was a clear effect of acclimation to lowered salinity on metal uptake, which may have important implications for metal accumulation modelling, biomonitoring, and toxicity studies. PMID- 12560171 TI - Impact of perfluorooctanoic acid on the structure of the zooplankton community in indoor microcosms. AB - There is presently, a substantial amount of information being gathered concerning the environmental risk associated with the perfluorooctanoic acid (PFOA) compound. The aim of this paper was to determine a 35 day community no observable effect concentration (NOEC(community)) or lowest observable effect concentration (LOEC) for freshwater zooplankton exposed to PFOA during a study in 30 l indoor aquatic microcosms. Some significant (P < 0.01) temporal fluctuations in zooplankton abundance were observed, however, a NOEC(community) could not be calculated. LOEC for various species varied between 10 and 70 mg l(-1). According to LOEC values, the tentative order of descriptors sensitivity was as follows: Daphnia magna > richness > or = Cyclops canthocamptus staphylinus > Cyclops diaptomus>total zooplankton > or = Rotifera sp. The long term ecological significance of these temporal fluctuations could not be determined in this study, however, the overall study cessation analysis showed that the structure of the ecosystem was changed from a more diverse community dominated by larger species towards a less diverse community dominated by smaller more and robust species (P < 0.05). Additional chronic toxicity testing should also be addressed since these compounds are so persistent and recalcitrant. PMID- 12560170 TI - Comparative effects and metabolism of two microcystins and nodularin in the brine shrimp Artemia salina. AB - The toxicity and metabolism of the cyanobacterial toxins microcystin-LR (MCLR), Dhb-microcystin-HtyR and nodularin were investigated in the cysts, nauplii and adults of the brine shrimp Artemia salina. The presence of the phase II detoxication system glutathione S-transferase (sGST) in these stages was shown using different substrates. Exposure of adult A. salina to the toxins led to an elevation of GST activity in vivo. All three toxins were conjugated to glutathione via GST, which has been shown as an initial step of microcystin and nodularin detoxication. PMID- 12560172 TI - The influence of ration size on copper homeostasis during sublethal dietary copper exposure in juvenile rainbow trout, Oncorhynchus mykiss. AB - The influence of ration size on homeostasis and sublethal toxicity of copper (Cu) was assessed in rainbow trout (Oncorhynchus mykiss) during dietary Cu exposure in synthetic soft water. A constant dietary dose of 0.24 micromol Cu per g fish per day as CuSO(4).5H(2)O was delivered via diets containing 15.75, 7.87, and 5.24 micromol Cu g(-1) fed at 1.5, 3.0, and 4.5% wet body weight daily ration, respectively. Juvenile rainbow trout showed clear effects of ration but not Cu on growth suggesting that growth is hardly a sensitive endpoint for detection of sublethal dietary Cu exposure. All Cu-exposed fish accumulated the same total metal load when expressed on a per fish basis. This suggests that differences in tissue and whole-body Cu concentrations among the treatments reflected the differences in the fish size rather than total Cu accumulation, and demonstrate that absorption and accumulation of Cu from the gut during dietary exposure are independent of the food quantity in which the Cu is delivered. Fish fed a high ration exhibited greater mass-specific unidirectional uptake of waterborne Cu than fish fed a low ration indicating an increased need for Cu for growth processes in rapidly growing fish. Stimulated excretion of Cu was indicated by greater Cu accumulation in the bile of the Cu-exposed fish. Branchial Na(+), K(+) ATPase was not affected by dietary Cu exposure or ration but gut Na(+), K(+) ATPase activities showed stimulatory effects of increasing ration but not of Cu exposure. The 96-h LC50 for waterborne Cu (range 0.17-0.21 micromol l(-1) (10.52 13.20 microg l(-1)) was the same in all treatment groups indicating that ration size was unimportant and that dietary Cu did not induce an increase in tolerance to waterborne Cu. Taken together these results suggest that the nutritional status, fish size, and growth rates should be considered when comparing whole body and tissue Cu concentration data for biomonitoring and risk assessment. Moreover, expressing the exposure as total metal dose rather than metal concentration in the diet is more appropriate. PMID- 12560174 TI - Openness in an insecure world. PMID- 12560173 TI - Stage sensitivity of medaka (Oryzias latipes) eggs and embryos to permethrin. AB - The effects of exposure to permethrin on gametes, fertilization and embryonic development were examined in medaka (Oryzias latipes). Following range finding (25, 50, 100, 200 or 300 microg/l) and duration of exposure (0, 120, 144, 168, 192, 216, or 240-h) assays, the relative sensitivity was studied when initiation of exposure to permethrin (100 microg/l, for 192-h) occurred at one of four different stages, i.e., unfertilized egg (0-h), late morula (5-h), early neurula (24-h), and early organogenesis (40-h). The later exposure interval proved the most sensitive. Also, differences were observed in rates of recovery in larvae initially affected following the earliest exposure treatment (0-h, gametes prior to fertilization). Permethrin (100 microg/l) did not affect fertilization success and no lethal effects were observed in embryos. Sublethal effects were primarily observed at hatch. Toxicity endpoints in larvae included: delayed swim bladder inflation; inability of hatchling to respond to stimulus; uncoordinated movements, myoskeletal defects and transient enlargement of gall bladder. These changes were characteristic for all hatchlings exposed to nominal concentrations of 50 microg/l. While certain of the above alterations were reversed within 72-h after hatching, lack of swim bladder inflation and inability to respond to stimuli were two features that persisted with significant incidences. Based on persistence of sublethal effects, results from this work indicate the importance of exposures to gametes and to embryos prior to water hardening. The approach taken herein may better reflect environmental risk conditions than assays limited to exposure of embryonated eggs. PMID- 12560175 TI - Positive response to US smallpox vaccine policy. PMID- 12560176 TI - Doubts cast on antimalarial drug. PMID- 12560178 TI - High hopes for HIV vaccine. PMID- 12560184 TI - Success for river blindness control campaign. PMID- 12560186 TI - The genomics revolution has arrived in infectious diseases. PMID- 12560187 TI - VanA-mediated high level glycopeptide resistance in MRSA. PMID- 12560188 TI - Epidemic meningococcal disease in sub-Saharan Africa--towards a sustainable solution? PMID- 12560189 TI - Miltefosine in visceral leishmaniasis. PMID- 12560190 TI - Nonoxynol-9 and sexually transmitted infections. PMID- 12560191 TI - Efficacy of polysaccharide pneumococcal vaccine in adults in more developed countries: the state of the evidence. AB - We review studies on the efficacy against disease caused by Streptococcus pneumoniae of the 23-valent polysaccharide pneumococcal vaccine in adult populations in the more developed countries. Meta-analyses of primary vaccine trials have attempted to reduce uncertainty from lack of power. Vaccine efficacy calculated from studies in South African gold-miners and in Papua New Guinea, with high attack rates and differing serotype patterns, cannot automatically be applied to more developed countries. Meta-analyses will overestimate a protective effect if this clinical heterogeneity is ignored. Meta-analyses limited to trials in the more developed setting show no protective effect against pneumococcal pneumonia and a non-significant protective effect against bacteraemia. Lack of a specific diagnosis limits the ability to detect a protective effect against pneumococcal pneumonia. Most, but not all, observational studies confirm a protective effect against bacteraemia. An effect on mortality in more developed countries has yet to be documented. PMID- 12560192 TI - Adenovirus: an increasingly important pathogen in paediatric bone marrow transplant patients. AB - Adenovirus is increasingly being recognised as a significant pathogen in children following bone marrow transplantation. The virus is endemic in the general paediatric population, and frequently causes severe disease in immunocompromised patients, especially children. Immune responses to adenovirus infection are not fully understood but T-cell responses appear to be important for recovery. Infections can affect a variety of organs with gastrointestinal and urinary tract diseases being the most common. When disseminated infection occurs, reported mortality rates are as high as 60%. The responses to treatment in immunocompromised patients have generally been disappointing. New molecular diagnostic techniques have meant that adenoviral infections can now be detected early, often before symptoms have developed. Clinicians now screen for adenovirus infection to allow early initiation of treatment. It is hoped that this approach, together with effective antiviral therapy, will reduce the deaths from this common virus in high-risk children. PMID- 12560193 TI - Septic thrombophlebitis with multiple pulmonary abscesses. PMID- 12560194 TI - Post-kala-azar dermal leishmaniasis. AB - Post-kala-azar dermal leishmaniasis (PKDL) is a complication of visceral leishmaniasis (VL); it is characterised by a macular, maculopapular, and nodular rash in a patient who has recovered from VL and who is otherwise well. The rash usually starts around the mouth from where it spreads to other parts of the body depending on severity. It is mainly seen in Sudan and India where it follows treated VL in 50% and 5-10% of cases, respectively. Thus, it is largely restricted to areas where Leishmania donovani is the causative parasite. The interval at which PKDL follows VL is 0-6 months in Sudan and 2-3 years in India. PKDL probably has an important role in interepidemic periods of VL, acting as a reservoir for parasites. There is increasing evidence that the pathogenesis is largely immunologically mediated; high concentrations of interleukin 10 in the peripheral blood of VL patients predict the development of PKDL. During VL, interferon gamma is not produced by peripheral blood mononuclear cells (PBMC). After treatment of VL, PBMC start producing interferon gamma, which coincides with the appearance of PKDL lesions due to interferon-gamma-producing cells causing skin inflammation as a reaction to persisting parasites in the skin. Diagnosis is mainly clinical, but parasites can be seen by microscopy in smears with limited sensitivity. PCR and monoclonal antibodies may detect parasites in more than 80% of cases. Serological tests and the leishmanin skin test are of limited value. Treatment is always needed in Indian PKDL; in Sudan most cases will self cure but severe and chronic cases are treated. Sodium stibogluconate is given at 20 mg/kg for 2 months in Sudan and for 4 months in India. Liposomal amphotericine B seems effective; newer compounds such as miltefosine that can be administered orally or topically are of major potential interest. Although research has brought many new insights in pathogenesis and management of PKDL, several issues in particular in relation to control remain unsolved and deserve urgent attention. PMID- 12560195 TI - The contribution of health-care services to a sound and sustainable malaria control policy. AB - HIV and AIDS, tuberculosis, and malaria, besides presenting a large mortality and morbidity burden in developing countries, are also responsible for poor economic development. In the past international agencies devoted resources and efforts to control malaria and other diseases without taking into account health-system performance and sustainability. Even assuming that the Global Fund to Fight AIDS, Tuberculosis, and Malaria (GFATM)--a recent international initiative--would provide the necessary funds, a poorly performing health-care system will not be able to use these funds optimally. Moreover, even if all interventions are cost effective, their impact on mortality and morbidity will only be marginal if access to proper care is not guaranteed. It is the responsibility of scientists and health managers to highlight to donor agencies the importance of an accessible and well functioning health-care system at all levels for the control of specific diseases. PMID- 12560196 TI - Child vaccination policies in Europe: a report from the Summits of Independent European Vaccination Experts. AB - Despite the proven safety and efficacy of vaccines, common vaccine-preventable diseases such as measles are not yet controlled in all European countries. This is largely due to three factors. First, vaccination systems differ widely throughout Europe and they vary between highly centralised and totally decentralised systems. Both have advantages and disadvantages, but without doubt they can all work locally. "Harmonisation" in this field is neither a prerequisite nor a guarantee for success. Second, perception of vaccination--and this includes education of the public--is most crucial. In this field the media play an important part, but their ability or will to communicate complicated scientific matters in an appropriate way to the public is often insufficient. Third, political will may be the single most important factor for success in vaccination. Only if the European Union comes up with and implements common vaccination goals with firm deadlines can the best health through vaccination of all Europeans be accomplished. The system as well as the schedule used would then be of minor importance. PMID- 12560198 TI - David Reddy--taking HIV therapy to new frontiers. Interview by Pam Das. PMID- 12560199 TI - Ecthyma gangrenosum and chronic lymphocytic leukaemia. PMID- 12560201 TI - Bacteria can't resist honey. PMID- 12560202 TI - Peptides that regulate food intake: orexin gene expression is increased during states of hypertriglyceridemia. AB - Previous reports implicate the orexins in eating and body weight regulation. This study investigated possible functional relationships between hypothalamic orexins and circulating hormones or metabolites. In situ hybridization and quantitative PCR were used to examine orexin expression in the perifornical hypothalamus (PF) of rats and mice on diets varying in fat content and with differential propensity toward obesity. The results showed that orexin gene expression was stimulated by a high-fat diet in close association with elevated triglyceride levels, suggesting a functional relationship between these measures. Results obtained in obesity-prone rats and mice revealed a similar increase in orexin in close relation to triglycerides. A direct test of this orexin-triglyceride link was performed with Intralipid, which increased PF orexin expression along with circulating triglycerides. Whereas PF galanin is similarly stimulated by dietary fat, double-labeling immunofluorescence studies showed that orexin and galanin neurons are anatomically distinct. This evidence suggests that the orexins, like galanin, are "fat-responsive" peptides that respond to circulating lipids. PMID- 12560203 TI - Cyclooxygenase-2 contributes to elevated renin in the early postnatal period in rats. AB - We asked whether cyclooxygenase (COX) activity controls the renin-angiotensin system in the postnatal period. During kidney development, renin peaked at postnatal days 0-1 at the mRNA, tissue protein [renal renin concentration (RRC)], and plasma renin concentration (PRC) levels and was widely expressed along preglomerular vessels. PRC and renin mRNA expression was elevated until weaning in the 4th postnatal week compared with adult rats. Renocortical COX-2 was restricted to Tamm-Horsfall protein-positive cells in the thick ascending limb of Henle's loop, and cortical COX-2 mRNA and protein expression were elevated along with PRC in the 2nd and 3rd postnatal weeks. In contrast, cortical COX-1 expression was constant, but medullary COX-1 expression increased eightfold from the 1st to 4th postnatal week. A COX-2-selective blocker, parecoxib, and a nonselective blocker, indomethacin, given in a period with COX-2 induction from postnatal day 6 to day 12, markedly decreased PRC, but not renin mRNA or RRC. Inhibition of angiotensin AT(1) receptors by candesartan from postnatal day 1 to day 5 increased COX-2 mRNA (2.5-fold), protein, and distribution, renin mRNA (7 fold) and PRC (20- to 70-fold), but had no influence on COX-1 mRNA. Thus, due to very low levels of expression, COX-2 is unlikely to be responsible for the birth peak of renin, but COX-2 activity supports renin secretion later in the suckling period. ANG II negatively feeds back on renocortical COX-2 expression in the 1st postnatal days with high activity of the renin system. We suggest that suckling in the rat is correlated to an enhanced, COX-2-mediated, secretory activity of renin-producing juxtaglomerular cells. PMID- 12560204 TI - Normal and glucocorticoid-induced development of the human small intestinal xenograft. AB - The aim of this study was to determine whether intestinal xenografts could recapitulate human in utero development by using disaccharidases as markers. Twenty-week-old fetal intestine was transplanted into immunocompromised mice and was followed. At 20-wk of gestation, the fetal human intestine was morphologically developed with high sucrase and trehalase but had low lactase activities. By 9-wk posttransplantation, jejunal xenografts were morphologically and functionally developed and were then monitored for 1 ng/mL per 10(6) cells per 72 hours), but AML-1A was not increased in MM cell lines that expressed less than 200 pg/mL MIP-1alpha. More importantly, the ratio of AML-1A to AML-1B mRNA levels was also increased in 3 of 3 highly purified myeloma cells from patients with MM who expressed increased amounts of MIP-1alpha. The ratio of AML-1A to AML-1B mRNA in patients with MM was 8-fold higher than in healthy controls. Transduction of AML-1B into the MM-derived MM.1S and ARH-77 cells totally blocked MIP-1alpha production, while AML-1A did not further increase the already high levels of MIP-1alpha produced by these cells. Taken together, these data demonstrate that in patients with MM who produce increased concentrations of MIP-1alpha, the relative level of AML-1B is significantly decreased compared with healthy controls. The data suggest that strategies that enhance AML-1B expression or decrease AML-1A in MM cells may be beneficial therapeutically. PMID- 12560230 TI - The low-frequency allele of the platelet collagen signaling receptor glycoprotein VI is associated with reduced functional responses and expression. AB - Interaction of platelets with collagen under conditions of blood flow is a multi step process with tethering via glycoprotein IbIXV (GPIbIXV) over von Willebrand factor, adhesion by direct interaction with the integrin GPIaIIa, and signaling via GPVI. GPVI can be specifically agonized by cross-linked collagen-related peptide (CRP-XL), which results in a signaling cascade very similar to that evoked by native collagen. The GPVI gene has 2 common alleles that differ by 3 replacements in the glycosylated stem and 2 in the cytoplasmic domain. We used CRP-XL to elucidate the variation in responses observed in platelet function in different individuals. We observed a 3-fold difference in the response to CRP-XL in platelet aggregation when comparing platelets from 10 high-frequency allele homozygotes with 8 low-frequency ones (2-way analysis of variance [ANOVA], P <.0001). The difference in functional responses was reflected in fibrinogen binding and in downstream signaling events as measured by tyrosine phosphorylation, the expression of P-selectin, and the binding of annexin V and the generation of thrombin on the platelet surface (2-way ANOVA, P <.001). Platelets homozygous for the low-frequency allele tended to be less able to form a thrombus on a collagen surface in flowing whole blood or in the platelet function analyzer-100 (t test, P =.065 and P =.061, respectively). The functional difference was correlated to a difference in total and membrane-expressed GPVI measured by monoclonal and polyclonal antibodies. This study demonstrates for the first time that platelet function may be altered by allelic differences in GPVI. PMID- 12560231 TI - A unique atrial primary cardiac lymphoma mimicking myxoma presenting with embolic stroke: a case report. AB - An immunocompetent 29-year-old male presented with an embolic stroke from an unusual primary cardiac lymphoma. The cardiac lesion consisted of a polypoid, left atrial, mural fibrin thrombus with anaplastic tumor cells lining the surface of the clot. Histologic, immunophenotypic, and molecular characterizations were consistent with a diagnosis of CD30+ large B-cell lymphoma with anaplastic cytology. While tumor emboli from invasive primary cardiac lymphomas have been reported, this noninvasive fibrin thrombus-associated lymphoma appears to be unique and previously unreported. PMID- 12560232 TI - Genetic evidence for convergence of c-Kit- and alpha4 integrin-mediated signals on class IA PI-3kinase and the Rac pathway in regulating integrin-directed migration in mast cells. AB - Mast cells play a critical role in host defense against a number of pathogens. Increased mast cell infiltration has been described in allergic asthma, in rheumatoid arthritis, and during helminthes infection. Despite the importance of mast cells in allergic disease and defense against infection, little is known about the mechanisms by which mast cells migrate to various tissues under steady state conditions or during infection or inflammation. Here, we show that activation of c-Kit by its ligand, stem cell factor (SCF), cooperates with alpha4 integrin in inducing directed migration of mast cells on fibronectin. A reduction in migration and activation of a small G protein, Rac, was observed in mast cells derived from class IA phosphoinositide-3 kinase (PI-3kinase)-deficient mice in response to SCF stimulation and in mast cells expressing the dominant-negative Rac (RacN17), as well as in mast cells deficient in the hematopoietic-specific small G protein, Rac2. In addition, a PI-3kinase inhibitor inhibited alpha4- as well as SCF-induced migration in a dose-dependent fashion. In contrast, a mitogen activated protein kinase (MAPK) inhibitor had little effect. Consistent with the pharmacologic results, abrogating the binding of the p85alpha subunit of class IA PI-3kinase to c-Kit also resulted in inhibition of SCF-induced migration on fibronectin. These genetic and biochemical data demonstrate that both c-Kit and alpha4 integrin signaling are linked to class IA PI-3kinase and Rac pathways and regulate integrin-directed (haptotactic) migration in mast cells. PMID- 12560233 TI - Tetrocarcin-A--induced ER stress mediates apoptosis in B-CLL cells via a Bcl-2- independent pathway. AB - Tetrocarcin-A (TC-A), an antibiotic agent isolated from actinomycetes, has recently been described to antagonize Bcl-2 functions, thereby sensitizing tumor cells to cell death signals under control of Bcl-2. In this study, we analyzed the direct proapoptotic effect of TC-A in the B-chronic lymphocytic leukemia (B CLL) model. We focused on the signal cascade triggered by TC-A in B-CLL cells and identified activated mitochondrial as well as endoplasmatic reticulum (ER) stress signals. The expression levels of known effector molecules mediating mitochondrial signaling, such as Bax and Bid, and the antagonistic molecule Bcl-2 did not influence sensitivity of B-CLL cells to TC-A. Furthermore, the molecular chaperone and sensor of ER stress, HSP70, though significantly up-regulated in B CLL cells undergoing TC-A-triggered apoptosis, was ineffective to exert its anti apoptotic function described in multiple cell death pathways. Autologous T cells of B-CLL patients were significantly less sensitive to TC-A as were also T cells from healthy donors when compared with their normal B-cell fraction. Furthermore, sensitivity of B-CLL cells to TC-A treatment in vitro was dependent neither on the expression levels of CD38-a prognostic factor for survival of B-CLL patients as well as for their response to therapy-nor on the clinical stage or pretreatment status of patients. From our data showing that TC-A induced a cell death pathway via ER stress preferentially in B cells and that it acted independently of important markers of drug sensitivity and of clinical markers, we conclude that TC-A might represent an attractive candidate drug for further evaluation in preclinical trials. PMID- 12560234 TI - Intranodal injection of semimature monocyte-derived dendritic cells induces T helper type 1 responses to protein neoantigen. AB - Dendritic cells (DCs) represent the most potent antigen-presenting cells of the immune system capable of initiating primary immune responses to neoantigens. Here we characterize the primary CD4 T-cell immune response to protein keyhole limpet hemocyanin (KLH) in 5 metastatic melanoma patients undergoing a tumor peptide based dendritic cell vaccination trial. Monocyte-derived dendritic cells displaying a semimature phenotype, as defined by surface markers, were loaded ex vivo with antigen and injected intranodally at weekly intervals for 4 weeks. All patients developed a strong and long-lasting delayed-type hypersensitivity reactivity to KLH, which correlated with the induction of KLH-dependent proliferation of CD4 T cells in vitro. Secondary in vitro stimulation with KLH showed significant increase in interferon-gamma and interleukin-2 (IL-2) but not IL-4, IL-5, nor IL-10 secretion by bulk T cells. On the single-cell level, most TH1 cells among in vitro-generated KLH-specific T-cell lines confirmed the preferential induction of a KLH-specific type 1 T helper immune response. Furthermore, the induction of KLH-specific antibodies of the IgG2 subtype may reflect the induction of a type 1 cytokine profile in vivo after vaccination. Our results indicate that intranodal vaccination with semimature DCs can prime strong, long-lasting CD4 T-cell responses with a TH1-type cytokine profile in cancer patients. PMID- 12560235 TI - In vitro identification of human pro-B cells that give rise to macrophages, natural killer cells, and T cells. AB - In this study we report the molecular and functional characterization of very early interleukin 7 receptor alpha (IL-7Ralpha)+-CD79a+CD19- B-cell progenitors, produced by human CD34+CD19-CD10- cord blood cells grown in the presence of stromal cells and cytokines. Purified IL-7Ralpha+CD79a+CD19- cells transcribed the B-lymphoid specific genes E2A, EBF, TdT, Rag-1, had initiated DJH rearrangements, but almost lacked Pax-5 mRNA. When exposed to appropriate environmental conditions, these cells repressed B-cell genes and completely differentiated into CD14+ macrophages, CD56+ natural killer cells, and CD4high T cells. Retention of the DJH rearranged genes in both CD14+ and CD56+ cells unambiguously demonstrates that early B-cell genes, expressed prior to Pax-5, can be activated in a multipotent human progenitor cell whose final fate, including in non-B lineages, is determined by external signals. PMID- 12560236 TI - Regulation of the human endothelial cell protein C receptor gene promoter by multiple Sp1 binding sites. AB - The human endothelial cell protein C receptor (hEPCR) is normally expressed by the endothelium of large blood vessels, but the molecular basis for its in vivo specificity is uncertain. In this study, DNaseI hypersensitive site mapping demonstrated the presence of a hypersensitive site in the 5' flanking region of the hEPCR gene in endothelial cells and certain transformed cells (HeLa and U937) known to express hEPCR in vitro. Conversely, this site was only weakly hypersensitive in HepG2 cells, cells which do not express hEPCR mRNA. Functional analysis of this 5' flanking region by in vivo dimethylsulfate footprinting in cultured endothelial cells identified multiple regions, containing high and low homology consensus Sp1 binding sequences, that were protected from methylation in endothelial cells. These sequences were not protected in HepG2 cells. Reporter gene analysis of this region in endothelial cells demonstrated the presence of promoter activity conferred by the proximal 572 bp but failed to identify a functional TATA-box. This promoter was inactive in HepG2 cells. Electrophoresis mobility shift assays using endothelial cell nuclear extracts identified Sp1 family proteins binding to sites that were protected during footprinting. Sp1 sites were identified in regions at -368, -232, -226, -201, -146, and -102 bp relative to the translation start site. With the exception of the site at -102 bp, each identified Sp1 binding site made a positive contribution to reporter gene expression, although no individual site was critically important. We conclude that transcription factor binding to Sp1 binding sites in the 5' flanking region is critical for normal hEPCR gene expression in endothelial cells. PMID- 12560237 TI - CCR5 deficiency decreases susceptibility to experimental cerebral malaria. AB - Infection of susceptible mouse strains with Plasmodium berghei ANKA (PbA) is a valuable experimental model of cerebral malaria (CM). Two major pathologic features of CM are the intravascular sequestration of infected erythrocytes and leukocytes inside brain microvessels. We have recently shown that only the CD8+ T cell subset of these brain-sequestered leukocytes is critical for progression to CM. Chemokine receptor-5 (CCR5) is an important regulator of leukocyte trafficking in the brain in response to fungal and viral infection. Therefore, we investigated whether CCR5 plays a role in the pathogenesis of experimental CM. Approximately 70% to 85% of wild-type and CCR5+/- mice infected with PbA developed CM, whereas only about 20% of PbA-infected CCR5-deficient mice exhibited the characteristic neurologic signs of CM. The brains of wild-type mice with CM showed significant increases in CCR5+ leukocytes, particularly CCR5+ CD8+ T cells, as well as increases in T-helper 1 (Th1) cytokine production. The few PbA-infected CCR5-deficient mice that developed CM exhibited a similar increase in CD8+ T cells. Significant leukocyte accumulation in the brain and Th1 cytokine production did not occur in PbA-infected CCR5-deficient mice that did not develop CM. Moreover, experiments using bone marrow (BM)-chimeric mice showed that a reduced but significant proportion of deficient mice grafted with CCR5+ BM develop CM, indicating that CCR5 expression on a radiation-resistant brain cell population is necessary for CM to occur. Taken together, these results suggest that CCR5 is an important factor in the development of experimental CM. PMID- 12560238 TI - Albumin-expressing hepatocyte-like cells develop in the livers of immune deficient mice that received transplants of highly purified human hematopoietic stem cells. AB - Rodent bone marrow cells can contribute to liver. If these findings are applicable to humans, marrow stem cells could theoretically be harvested from a patient and used to repair his/her damaged liver. To explore this potential, CD34(+) or highly purified CD34(+)CD38(-)CD7(-) human hematopoietic stem cells from umbilical cord blood and bone marrow were transplanted into immunodeficient mice. One month after transplantation, carbon tetrachloride (CCl(4)) was administered into the mice to induce liver damage and hepatocyte proliferation. Mice were analyzed in comparison with CCl(4)-injured mice that did not receive transplants and noninjured controls that received transplants with the same stem cell populations, one month after liver damage. Human-specific albumin mRNA and protein were expressed in the mouse liver and human albumin was detected in the serum of mice that had received CCl(4) injury. Human alpha-fetoprotein was never expressed, but in some mice, human cytokeratin 19 was expressed, which may indicate bile duct development in addition to the albumin-secreting hepatocyte like cells. Human albumin was not expressed in the starting stem cell populations in injured mice that did not receive transplants nor in noninjured mice that had received transplants of human stem cells. Human albumin expression was detected only in CCl(4)-treated mice that received transplants of human stem cells, and recovery was increased by administration of human hepatocyte growth factor 48 hours after the CCl(4)-mediated liver injury. Our studies provide evidence that human "hematopoietic" stem/progenitor cell populations have the capacity to respond to the injured liver microenvironment by inducing albumin expression. PMID- 12560239 TI - The in vivo profile of transcription factors during neutrophil differentiation in human bone marrow. AB - In vivo distribution of myeloid transcription factors during granulopoiesis was investigated by Northern and Western blotting in 3 neutrophil precursor populations from human bone marrow: immature (myeloblasts [MBs] and promyelocytes [PMs]); intermediate mature (myelocytes [MCs] and metamyelocytes [MMs]); and mature neutrophil cells (band cells [BCs] and segmented neutrophil cells [SCs]). Nonneutrophil cells were removed with magnetic-bead-coupled antibodies against CD2, CD3, CD14, CD19, CD56, CD61, glycophorin-A, and CD49d (BCs/SCs) before RNA and protein extraction. Polymorphonuclear neutrophils (PMNs) from peripheral blood depleted with anti-CD49d antibodies were also included. Expression of acute myeloid leukemia 1b (AML-1b), c-myb, GATA-1, and CCAAT/enhancer binding protein gamma (C/EBP-gamma) was seen primarily in MBs/PMs, and little expression was found in more mature cells. The level of C/EBP-alpha was constant in the bone marrow-derived cells and decreased in PMNs. C/EBP-epsilon was found primarily in MCs/MMs and was almost absent in more mature cells. Expression of C/EBP-beta, C/EBP-delta, and C/EBP-zeta was observed from the MC/MM stage onward, with peak levels in the most mature cells. The amount of PU.1 increased throughout maturation whereas the level of Elf-1 reached a nadir in MCs/MMs The PU.1 coactivator c-jun and c-jun's dimerization partner c-fos were both detectable in MCs/MMs and increased in amount with maturity. CCAAT displacement protein (CDP) was found at comparable levels at all stages of differentiation. This demonstrates a highly individualized expression of the transcription factors, which can form the basis for the heterogeneous expression of granule proteins during granulopoiesis and cell cycle arrest in metamyelocytes. PMID- 12560240 TI - Altered erythrocyte endothelial adherence and membrane phospholipid asymmetry in hereditary hydrocytosis. AB - The risk for thrombosis is increased in patients with hereditary hydrocytosis, an uncommon variant of hereditary stomatocytosis. Erythrocytes from 2 patients with hydrocytosis were studied to gain insight into the mechanism of thrombosis in this disorder. Erythrocytes demonstrated abnormal osmotic scan ektacytometry and decreased erythrocyte filtration rates. There was also a mild increase in adherence of erythrocytes to endothelial monolayers in a micropipette assay. Adhesion of erythrocytes to the subendothelial matrix proteins thrombospondin and laminin, however, was not significantly increased. Percentages of hydrocytosis erythrocytes and reticulocytes with phosphatidylserine exposed on the outer surfaces were increased in both patients compared with healthy controls, indicating altered membrane phospholipid asymmetry. Increased phosphatidylserine exposure accelerating thrombin-forming processes has been proposed as a mechanism for thrombosis in sickle cell disease and beta-thalassemia and may play a similar role in hereditary hydrocytosis. PMID- 12560242 TI - Global down-regulation of HOX gene expression in PML-RARalpha + acute promyelocytic leukemia identified by small-array real-time PCR. AB - Acute promyelocytic leukemia (APL) is associated with a reciprocal and balanced translocation involving the retinoic acid receptor-alpha (RARalpha). All-trans retinoic acid (ATRA) is used to treat APL and is a potent morphogen that regulates HOX gene expression in embryogenesis and organogenesis. HOX genes are also involved in hematopoiesis and leukemogenesis. Thirty-nine mammalian HOX genes have been identified and classified into 13 paralogous groups clustered on 4 chromosomes. They encode a complex network of transcription regulatory proteins whose precise targets remain poorly understood. The overall function of the network appears to be dictated by gene dosage. To investigate the mechanisms involved in HOX gene regulation in hematopoiesis and leukemogenesis by precise measurement of individual HOX genes, a small-array real-time HOX (SMART-HOX) quantitative polymerase chain reaction (PCR) platform was designed and validated. Application of SMART-HOX to 16 APL bone marrow samples revealed a global down regulation of 26 HOX genes compared with normal controls. HOX gene expression was also altered during differentiation induced by ATRA in the PML-RARalpha(+) NB4 cell line. PML-RARalpha fusion proteins have been reported to act as part of a repressor complex during myeloid cell differentiation, and a model linking HOX gene expression to this PML-RARalpha repressor complex is now proposed. PMID- 12560241 TI - Distinct contributions of TNF and LT cytokines to the development of dendritic cells in vitro and their recruitment in vivo. AB - TNF/LTalpha/LTbeta (tumor necrosis factor/lymphotoxin-alpha/lymphotoxin-beta) triple knockout (KO) mice show a significant reduction of dendritic cell (DC) number in the spleen, presumably due to defective recruitment and/or production. To distinguish between these possibilities, DCs were generated from bone marrow (BM) cultures prepared from wild-type (wt) and mutant mice in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL 4). The yield of CD11c(+) major histocompatibility complex (MHC) class II(+) DCs generated from TNF/LTalpha/LTbeta(-/-) BM culture was significantly reduced compared with wt BM culture. In order to further dissect the individual pathways responsible for defective DC properties observed in TNF/LTalpha/LTbeta(-/-) mice, the panel of TNF/LT ligand and receptor single KO mice were used. The production of DCs from BM culture was significantly reduced in TNF(-/-) and TNF receptor (TNFR) p55(-/-) mice, but normal in LTalpha(-/-), LTbeta(-/-), LTbetaR(-/-) mice. Recombinant TNF (rTNF) exogenously added to TNF/LTalpha/LTbeta(-/-) BM cultures could reverse this defect, and blocking antibodies showed partial effect on BM cultures of wt mice. Conversely, numbers of mature DCs in spleen were significantly decreased in LTalpha(-/-), LTbeta(-/-), LTbetaR(-/-) mice, but not in TNF(-/-) and TNFRp55(-/-) mice. These results reveal 2 distinct contributions of TNF/LT cytokines. First, TNF acting through TNF receptor is involved in the development/maturation of DCs in BM progenitor cultures, but this function appears to be redundant in vivo. Second, the microenvironment in peripheral lymphoid organs associated with LTalpha/LTbeta-LTbetaR signaling and chemokine production is critical for recruitment efficiency of DCs, and this pathway is indispensable. PMID- 12560243 TI - Translocation of the IgH locus is nearly ubiquitous in multiple myeloma as detected by immuno-FISH. PMID- 12560244 TI - Pure red-cell aplasia due to parvovirus B19 infection in a patient treated with alemtuzumab. PMID- 12560245 TI - Thrombocytopenia following treatment with platelet glycoprotein IIb/IIIa inhibitors. PMID- 12560246 TI - On the use of DNA methylation inhibitors and the reversal of transcriptional silencing. PMID- 12560247 TI - IVIG induces dose-dependent amelioration of ITP in rodent models. PMID- 12560248 TI - Helicobacter pylori and idiopathic thrombocytopenic purpura in children. PMID- 12560249 TI - Autoimmune complexes in human serum in presence of therapeutic amounts of intravenous immunoglobulins. PMID- 12560250 TI - Therapeutic thrombolysis for acute ischaemic stroke. PMID- 12560251 TI - "Schools and hospitals" for "education and health". PMID- 12560252 TI - Old drugs for new bugs. PMID- 12560253 TI - The BetterCare judgment--a challenge to health care. PMID- 12560254 TI - Closing the digital divide. PMID- 12560255 TI - Climbie inquiry recommends national agency for children. PMID- 12560257 TI - Egg or sperm donation children will be entitled to more information. PMID- 12560260 TI - Little evidence for effectiveness of scientific peer review. PMID- 12560261 TI - Link found between Agent Orange and chronic lymphocytic leukaemia. PMID- 12560263 TI - Swiss parliament may try to ban "suicide tourism". PMID- 12560264 TI - UK lags behind rest of Europe in reducing deaths from heart disease. PMID- 12560265 TI - Social costs of smoking are triple those of illicit drugs. PMID- 12560266 TI - Shipman proposals will alter general practice profoundly. PMID- 12560267 TI - England short of 970 GPs. PMID- 12560268 TI - Italian courts find 175 doctors and managers guilty of fraud. PMID- 12560269 TI - High Court strikes down Merck's patent on alendronate. PMID- 12560270 TI - Preparing for the worst. PMID- 12560271 TI - Detection of Alzheimer's disease and dementia in the preclinical phase: population based cohort study. AB - OBJECTIVES: To evaluate a simple three step procedure to identify people in the general population who are in the preclinical phase of Alzheimer's disease and dementia. DESIGN: Three year population based cohort study. SETTING: Kungsholmen cohort, Stockholm, Sweden. PARTICIPANTS: 1435 people aged 75-95 years without dementia. ASSESSMENTS: Single question asking about memory complaints, assessment by mini-mental state examination, and neuropsychological testing. MAIN OUTCOME MEASURE: Alzheimer's disease and dementia at three year follow up. RESULTS: None of the three instruments was sufficiently predictive of Alzheimer's disease and dementia when administered separately. After participants had been screened for memory complaints and global cognitive impairment, specific tests of word recall and verbal fluency had positive predictive values for dementia of 85-100% (95% confidence intervals range from 62% to 100%). However, only 18% of future dementia cases were identified in the preclinical phase by this three step procedure. Memory complaints were the most sensitive indicator of Alzheimer's disease and dementia in the whole population, but only half the future dementia cases reported memory problems three years before diagnosis. CONCLUSION: This three step procedure, which simulates what might occur in clinical practice, has a high positive predictive value for dementia, although only a small number of future cases can be identified. PMID- 12560272 TI - Fetal growth and subsequent risk of breast cancer: results from long term follow up of Swedish cohort. AB - OBJECTIVE: To investigate whether size at birth and rate of fetal growth influence the risk of breast cancer in adulthood. DESIGN: Cohort identified from detailed birth records, with 97% follow up. SETTING: Uppsala Academic Hospital, Sweden. PARTICIPANTS: 5358 singleton females born during 1915-29, alive and traced to the 1960 census. MAIN OUTCOME MEASURES: Incidence of breast cancer before (at age <50 years) and after (> or = 50 years) the menopause. RESULTS: Size at birth was positively associated with rates of breast cancer in premenopausal women. In women who weighed > or =4000 g at birth rates of breast cancer were 3.5 times (95% confidence interval 1.3 to 9.3) those in women of similar gestational age who weighed <3000 g at birth. Rates in women in the top fifths of the distributions of birth length and head circumference were 3.4 (1.5 to 7.9) and 4.0 (1.6 to 10.0) times those in the lowest fifths (adjusted for gestational age). The effect of birth weight disappeared after adjustment for birth length or head circumference, whereas the effects of birth length and head circumference remained significant after adjustment for birth weight. For a given size at birth, gestational age was inversely associated with risk (P=0.03 for linear trend). Adjustment for markers of adult risk factors did not affect these findings. Birth size was not associated with rates of breast cancer in postmenopausal women. CONCLUSIONS: Size at birth, particularly length and head circumference, is associated with risk of breast cancer in women aged <50 years. Fetal growth rate, as measured by birth size adjusted for gestational age, rather than size at birth may be the aetiologically relevant factor in premenopausal breast cancer. PMID- 12560273 TI - Controlled trial of effect of documented cardiovascular risk scores on prescribing. PMID- 12560274 TI - Discrepancies between patients' assessments of outcome: qualitative study nested within a randomised controlled trial. PMID- 12560275 TI - Influence of the third generation pill controversy on prescriptions for oral contraceptives among first time users: population based study. PMID- 12560276 TI - Cholesterol lowering drugs and risk of age related maculopathy: prospective cohort study with cumulative exposure measurement. PMID- 12560277 TI - Mortality from cardiovascular disease more than 10 years after radiotherapy for breast cancer: nationwide cohort study of 90 000 Swedish women. PMID- 12560278 TI - Thromboembolism associated with the new contraceptive Yasmin. PMID- 12560280 TI - Chronic cough. PMID- 12560279 TI - Setting standards based on patients' views on access and continuity: secondary analysis of data from the general practice assessment survey. AB - OBJECTIVES: To examine patients' views on access and continuity in general practice to derive quality standards. DESIGN: Secondary analysis of data from general practice research studies and routine quality assessment activities undertaken by practices and primary care trusts. SETTING: General practice. PARTICIPANTS: General practice patients. RESULTS: Satisfactory standards of access were next day appointments with general practitioners and a 6-10 minute wait for consultations to begin. A satisfactory level of continuity was seeing the same general practitioner "a lot of the time." Standards varied with the analytic method used and by sociodemographic group. CONCLUSIONS: Standards expected by patients in primary care can be derived from linked report-assessment pairs. Patients may have expectations of access that are in excess of government targets. Patients also have high expectations of continuity of care. It is unclear the degree to which such standards are reliable or valid, how conflicts between access and continuity should be resolved, or how these standards relate to other priorities of patients such as high quality interpersonal care. PMID- 12560281 TI - Pathogenesis of sepsis: new concepts and implications for future treatment. PMID- 12560282 TI - Low dose methotrexate and bone marrow suppression. PMID- 12560283 TI - ABC of learning and teaching in medicine. Curriculum design. PMID- 12560284 TI - Assisted suicide and euthanasia in Switzerland: allowing a role for non physicians. PMID- 12560285 TI - Monitoring mortality rates in general practice after Shipman. PMID- 12560286 TI - Getting married in China: pass the medical first. PMID- 12560287 TI - Volume of procedures and outcome of treatment. NHS needs to understand relation more effectively. PMID- 12560288 TI - Electronic tagging of people with dementia. Devices may be preferable to locked doors. PMID- 12560289 TI - Squamous cell carcinomas of the head and neck. Oral care for patients with cancer needs more than lip service. PMID- 12560290 TI - Screening for aortic aneurysm. Human cost should not be dismissed. PMID- 12560291 TI - Haemophilus influenzae type b epiglottitis. Occasional cases will present. PMID- 12560296 TI - Tracheal intubation: unconsciousness, analgesia and muscle relaxation. PMID- 12560297 TI - Rapid intervention for an episode of malignant hyperthermia. PMID- 12560299 TI - A combination of alfentanil-lidocaine-propofol provides better intubating conditions than fentanyl-lidocaine-propofol in the absence of muscle relaxants. AB - PURPOSE: To compare the ease of tracheal intubation without the use of muscle relaxants following an alfentanil-lidocaine-propofol sequence vs a fentanyl lidocaine-propofol sequence. CLINICAL FEATURES: In 80 ASA I and II adult patients undergoing elective laparoscopic surgery, we compared the intubating conditions following alfentanil 20 microg x kg(-1), lidocaine 1.5 mg x kg(-1), propofol 3 mg x kg(-1) (Group I; n = 40) vs fentanyl 2 microg x kg(-1), lidocaine 1.5 mg x kg( 1), propofol 3 mg x kg(-1) (Group II; n = 40). The intubating conditions were scored by jaw relaxation, vocal cord position and response to intubation, as well as by blood pressure and heart rate changes. The intubating conditions were good or excellent in 95% of patients in Group I vs 62.5% of patients in Group II (P < 0.05). Blood pressure decreased from a preinduction value of 86 +/- 13 mmHg to 72 +/- 28 mmHg and 74 +/- 19 mmHg in Group I, and from 85 +/- 12 mmHg to 78 +/- 15 mmHg and 78 +/- 12 mmHg in Group II, one and five minutes following intubation (P < 0.05). This drop in blood pressure was not different between the two groups. CONCLUSION: An alfentanil-lidocaine-propofol sequence offers significantly better intubating conditions than a fentanyl-lidocaine-propofol sequence in healthy adult patients. PMID- 12560298 TI - Propofol - not thiopental or etomidate - with remifentanil provides adequate intubating conditions in the absence of neuromuscular blockade. AB - PURPOSE: Administration of remifentanil followed by propofol provides adequate conditions for tracheal intubation without muscle relaxants. Other hypnotic drugs have not been thoroughly investigated in this regard. Intubating conditions with remifentanil followed by propofol, thiopentone or etomidate are compared in this study. METHODS: In a randomized, double-blind study 45 healthy males were assigned to one of three groups (n = 15). After iv atropine, remifentanil 3 microg x kg(-1) were injected over 90 sec followed by propofol 2 mg x kg(-1) (Group I), thiopentone 6 mg x kg(-1) (Group II) or etomidate 0.3 mg x kg(-1) (Group III). Ninety seconds after the administration of the hypnotic agent, laryngoscopy and intubation were attempted. Intubating conditions were assessed as excellent, good or poor on the basis of ease of ventilation, jaw relaxation, position of the vocal cords, and patient response to intubation and slow inflation of the endotracheal tube cuff. RESULTS: One patient in Group I, three patients in Group II and five patients in Group III could not be intubated on the first attempt. Clinically acceptable intubating conditions were observed in 93.3%, 66.7%, 40.0% of patients in Groups I, II and III, respectively. Overall conditions at intubation were significantly (P < 0.05) better, and the frequency of excellent conditions was significantly (P < 0.05) higher in the propofol group compared with the thiopentone and etomidate groups. No patient was treated for hypotension or bradycardia. CONCLUSION: Propofol 2 mg x kg(-1) was superior to thiopentone 6 mg x kg(-1) and etomidate 0.3 mg x kg(-1) for tracheal intubation when combined with remifentanil 3 microg x kg(-1) and no muscle relaxant. PMID- 12560300 TI - Muscle relaxation and increasing doses of propofol improve intubating conditions. AB - PURPOSE: Muscle relaxants and anesthetics are usually associated during intubation. However, their relative role to facilitate the process is not clearly defined. This study was designed to determine, during intubation: i). the relative role of anesthetics and atracurium-induced neuromuscular block and; ii). the effect of different doses of propofol in the presence of complete muscle block. METHODS: Patients were randomized to four groups and received fentanyl and a standardized anesthetic procedure. Patients from groups high (H; n = 45), medium (M; n = 48) and low (L; n = 47) received 2.5 mg x kg(-1), 2.0 mg x kg(-1), and 1.5 mg x kg(-1) of propofol respectively. Atracurium (0.5 mg x kg(-1)) was then injected and tracheal intubation performed once complete block was achieved at the orbicularis oculi. Patients from group without atracurium (WA; n = 20) received propofol as in group H. Intubation was performed at the expected onset time of action of atracurium. RESULTS: Using the same dose of propofol, the incidence of good or excellent intubating conditions was 35% without atracurium and 95% with atracurium (P < 0.0001). In patients receiving atracurium, clinically acceptable intubating conditions were more frequently achieved in groups receiving the highest propofol doses (group H or M vs group L; P < 0.03). CONCLUSION: Our study confirms the interaction between anesthesia and muscle relaxation to produce adequate intubating conditions. In the conditions described, intubating conditions were more dependent on atracurium-induced neuromuscular blockade than on anesthetics, but both atracurium and propofol improved intubating conditions. PMID- 12560301 TI - Warmed diluent speeds dantrolene reconstitution. AB - PURPOSE: To define more completely the aqueous solubility of dantrolene in order to devise faster reconstitution techniques for use during malignant hyperthermia episodes. METHODS: To determine Beer's law compliance and the extinction coefficient, we measured the spectrophotometric absorbance at 385 nm of known dantrolene solutions. We added small aliquots of sterile water USP (pH 5.3; 15-40 degrees C) or buffered water (20 degrees C; pH 6.8-9.1) to dantrolene, mechanically agitated and filtered the solutions, and spectrophotometrically determined concentration. To simulate clinical reconstitution conditions, we added sterile water, 60 mL, at temperatures between 15 and 40 degrees C to dantrolene vials and measured the manual shaking time needed to create a) a suspension of small particles, and b) a clear solution. RESULTS: A plot of UV-vis absorbance at 385 nm vs dantrolene concentration was linear and went through the origin; the extinction coefficient is 16.1 mM(-1). At 20 degrees C, dantrolene is nearly insoluble below pH 8.8. Dantrolene is 2.8 times more soluble in 0.1 M THAM (tris-(hydroxymethyl)aminomethane) than in sterile water at pH 9.1. Dantrolene is 6.7 times more soluble in 40 degrees C than in 20 degrees C water at pH 9.5 (the pH of reconstituted dantrolene). Under clinical conditions, water temperature altered the time to create a clear solution but not a suspension (60 sec). CONCLUSION: Diluting dantrolene with 40 degrees C water rather than operating room temperature water (20 degrees C or below) would speed dantrolene reconstitution. PMID- 12560302 TI - Successful implementation of perioperative beta-blockade utilizing a multidisciplinary approach. AB - PURPOSE: To describe how we implemented a protocol for perioperative beta blockade in patients with or at risk of coronary artery disease (CAD) undergoing major non-cardiac surgery and to present our results. METHODS: After institutional approval, from May 1999 to April 2001, patients with surgical and medical indications (CAD as indicated by previous myocardial infarction, typical angina or atypical angina with a positive stress test or at least two risk factors for CAD: age 65 yr, hypertension, smoking, high cholesterol, diabetes mellitus) for perioperative beta-blockade were identified preoperatively by anesthesiology and referred to the General Internal Medicine Service (MED). MED initiated patients on outpatient beta-blockers. The intraoperative anesthetic management was left to the discretion of the anesthesiologist. In the postanesthesia care unit (PACU), patients received iv metoprolol according to hemodynamic criteria. Postoperatively, patients were followed by MED for adverse cardiac events. RESULTS: Sixty-nine patients received perioperative beta blockade. Preoperatively, 60% were started on metoprolol, 39% on atenolol and 1% on propranolol. In PACU, 42%, 9% and 38% of patients were given iv metoprolol 0, 5 and 10 mg respectively. One patient was given glycopyrrolate in the PACU for bradycardia and none received vasoactive or inotropic agents. Three patients (4.3%) had postoperative cardiac events. CONCLUSIONS: With close collaboration between anesthesiologists, internists, PACU nurses and family physicians, a strategy for perioperative beta-blockade was implemented successfully in patients with cardiac risks. Beta-blockade was associated with few side effects and morbidities. PMID- 12560303 TI - Both the OxyArm and Capnoxygen mask provide clinically useful capnographic monitoring capability in volunteers. AB - PURPOSE: To compare the capnography monitoring performance of the new OxyArm (OA) with the Capnoxygen mask (CM), a conventional oxygen mask with a carbon dioxide sampling port. METHODS: Eleven healthy volunteer adult subjects underwent capnographic monitoring (in a non-randomized, un-blinded crossover study) at baseline and while receiving oxygen at seven different flow rates (0.5, 1.0, 2.0, 4.0, 6.0, 8.0, and 10 L x min(-1)), applied first with the CM and then with the OA. RESULTS: Both the OA and CM produced acceptable capnographs with consistent waveforms. The measured end-tidal (ET) CO(2) was equivalent for the two devices at all seven oxygen flow rates. On average, the ETCO(2) measured with the OA was about 2 mmHg greater than that of the CM. Regression analysis showed an inverse relationship between oxygen therapy flow rate and measured ETCO(2) whereby the measured value of CO(2) decreased as the oxygen flow rate was increased (P < 0.001). Both the CM and OA produced consistent measurements of ETCO(2) as illustrated by their reliability coefficients, 0.95 and 0.86 respectively. The biggest source of variation in measured CO(2) for both devices was inter-subject differences, followed by variable oxygen flow rates. CONCLUSIONS: This study suggests that the OA and CM can prove useful for respiratory monitoring and oxygen delivery in spontaneously breathing volunteers, and the OA could potentially be used as an alternative to the conventional methods of oxygen delivery and CO(2) sampling in patients. PMID- 12560304 TI - Best evidence in anesthetic practice: prevention: epidural anesthesia and analgesia does not reduce 30-day all-cause mortality and major morbidity after abdominal surgery. PMID- 12560305 TI - The addition of droperidol or clonidine to epidural tramadol shortens onset time and increases duration of postoperative analgesia. AB - PURPOSE: To compare tramadol alone and the combinations of either tramadol clonidine or tramadol-droperidol with regard to analgesic and adverse effects. METHODS: After Ethic's Committee approval and patient informed consent were obtained, epidural catheters were inserted preoperatively at the L(3-4) interspace in 90 ASA physical status I-II adult patients undergoing lower abdominal surgery. Anesthesia was standardized. Patients were randomly assigned to one of three groups. Group I (T) patients received tramadol 75 mg, Group II (TD) patients received tramadol 75 mg plus droperidol 2.5 mg, and Group III (TC) patients received tramadol 75 mg plus clonidine 150 microg in a total volume of 10 mL administered as a single epidural injection in the postanesthesia care unit. The onset time of analgesia and duration of analgesia, visual analogue pain scores, sedation, nausea scores, vital signs and side effects were recorded. RESULTS: Duration of analgesia was similar in both the TD and TC groups, and significantly longer than in the T group (P < 0.001). Group TC patients displayed a significant increase in sedation scores and decrease in blood pressure and heart rate when compared with other groups (P < 0.001). No adverse effects were observed in Group TD, while nausea scores were high in both the T and TC groups (P < 0.001). Pain score, respiration rate, and SpO(2) values were similar in all study groups. CONCLUSION: We conclude that epidural tramadol in combination with droperidol or clonidine prolongs the duration of analgesia; however, droperidol appears to be a better alternative when adverse effects and antiemetic properties are taken into consideration. PMID- 12560306 TI - Intradermal sufentanil does not improve lidocaine-induced local anesthesia. AB - PURPOSE: Peripheral opioid receptors may result in antinociceptive effects when occupied by opioids. This study examined intradermally injected sufentanil (S), a highly lipid soluble opioid, administered with and without lidocaine (L), in a thermal pain model. METHODS: Nine volunteers were instructed on the method of magnitude estimation of pain before undergoing baseline testing with seven seconds thermal stimuli between 44 and 52 degrees C, delivered by a contact thermal stimulator at five cutaneous forearm sites. Then, four sites randomly received injections of equal volumes (0.1 mL) of either normal saline (NS), lidocaine 0.5% (L), sufentanil 0.75 microg (S), lidocaine 0.5% plus sufentanil 0.75 microg (L+S), and one site was not injected and served as reference (REF). Testing was repeated at six, 30, 60, 90, 120, and 150 min following injection. The pain elicited by each stimulus was normalized to the subject's response to the 50 degrees C stimulus at the REF site. RESULTS: Baseline testing showed small (P = ns) differences in pain scores. At six minutes, the lidocaine sites (L, L+S) had pain scores that were mean 83% (range 78-88%) lower than the other sites (P < 0.05), but there was no difference between the L and L+S sites or between the S and NS or REF sites. At 30 and 60 min these pain scores were mean 38% (29-44%) and 20% (8-30%) less than at the REF, NS, and S sites (P = ns). At 90 min and later times, the pain scores had returned to baseline. CONCLUSIONS: These results suggest that intradermal sufentanil alone has no analgesic effect. Further, in combination with lidocaine, sufentanil does neither potentiate nor prolong the analgesic effect of lidocaine. PMID- 12560307 TI - Images in anesthesia: headache caused by pneumocephalus following inadvertent dural puncture during epidural space identification: is it time to abandon the loss of resistance to air technique? PMID- 12560308 TI - Peripartum cardiomyopathy and thromboembolism; anesthetic management and clinical course of an obese, diabetic patient. AB - PURPOSE: To describe the anesthetic management and clinical course of a patient with peripartum cardiomyopathy. We highlight the frequent occurrence of thromboembolic morbidity in this group of parturients, emphasizing the need for early consideration of prophylactic anticoagulation. CLINICAL FEATURES: A 38-yr old, diabetic, obese parturient was admitted with pulmonary edema and severe orthopnea at 31 weeks gestation. The respiratory rate was 44 breaths x min(-1), blood pressure 110/70 mmHg, pulse 120 beats x min(-1) and rales were heard in both lung fields. The diagnosis of peripartum cardiomyopathy was made based on sinus tachycardia with no evidence of ischemia on the electrocardiogram, and global left ventricular hypokinesis with an ejection fraction of 40-45% noted on transthoracic echocardiography. Cesarean delivery was planned to improve maternal respiratory status and hemodynamics. General anesthesia with invasive monitoring was planned, and surgery and anesthesia proceeded uneventfully. Less than 24 hr postoperatively, she sustained a thrombotic cerebral infarct leaving her hemiparetic and dysarthric. Subsequent investigations revealed a thrombophilic state due to elevated anticardiolipin antibody. CONCLUSION: General anesthesia is an acceptable option in parturients with heart failure secondary to cardiomyopathy. Thromboembolic complications are common, and early consideration should be given to prophylactic anticoagulation. PMID- 12560309 TI - Cerebrovascular reactivity to carbon dioxide is preserved during hypocapnia in children anesthetized with 1.0 MAC, but not with 1.5 MAC desflurane. AB - PURPOSE: Maintenance of cerebrovascular reactivity to CO(2) (CCO(2)R) is important during neurosurgical anesthesia. This study was designed to determine the effect of different desflurane concentrations on CCO(2)R in children. METHODS: Children undergoing urological surgery were enrolled. Anesthesia was induced with sevoflurane in air/oxygen. After intubation, sevoflurane was switched to desflurane. Analgesia was provided with an epidural neuraxial block. Mechanical ventilation was adjusted to an initial EtCO(2) of 30 mmHg. Exogenous CO(2) was used to achieve an EtCO(2) of 40 and 50 mmHg. Patients were randomized to the sequence of desflurane concentration (1.0 and 1.5 MAC) and the EtCO(2). Transcranial Doppler was used to measure middle cerebral artery blood flow velocity (Vmca). Five minutes were allowed to reach steady state after each change in EtCO(2) and 15 min after changing the desflurane concentration. RESULTS: Sixteen patients were studied. The mean age and weight were 3.5 +/- 1.5 yr and 14.4 +/- 3.1 kg, respectively. Mean arterial pressure remained stable throughout the study, while at an EtCO(2) of 50 mmHg, heart rate decreased at both desflurane concentrations (P < 0.05). At 1.0 MAC, Vmca increased from 30 to 40 mmHg (P < 0.05), but not from 40 to 50 mmHg EtCO(2). At 1.5 MAC, Vmca increased between 30 and 50 mmHg (P < 0.05). CONCLUSION: CCO(2)R is preserved during hypocapnia in children anesthetized with 1.0 MAC, but not with 1.5 MAC desflurane. The lack of further increase in Vmca at higher EtCO(2) concentrations implies that desflurane may cause significant cerebral vasodilatation in children. This may have important implications in children with reduced intracranial compliance. PMID- 12560310 TI - Magnesium potentiates neuromuscular blockade with cisatracurium during cardiac surgery. AB - PURPOSE: Magnesium potentiates the effect of nondepolarizing neuromuscular blocking agents. It is used in cardiac anesthesia to prevent hypertension and arrhythmias. This study was performed to measure the interaction between magnesium and cisatracurium in cardiac surgery. METHODS: Twenty patients scheduled for elective cardiac surgery were randomly assigned to receive magnesium sulfate (70 mg x kg(-1) at induction followed by 30 mg x kg(-1) x hr( 1)) or placebo. The ulnar nerve was stimulated and the electromyographic response of the adductor pollicis was measured. Cisatracurium 0.1 mg x kg(-1) was given at induction, followed by 0.05 mg x kg(-1) when the first twitch in the train-of four reached 25%. RESULTS: Ionized magnesium was 1.32 +/- 0.24 mmol x L(-1) in the treatment group vs 0.47 +/- 0.4 mmol x L(-1) in the control group. Duration of action of the intubating dose was longer in the magnesium group (74 +/- 20 min) than in the placebo group (42 +/- 6 min, P = 0.0001). Duration of the first maintenance dose was 69 +/- 16 min in the magnesium group vs 35 +/- 7 min in the placebo group (P = 0.0001). Total dose of cisatracurium administered throughout surgery was 0.19 +/- 0.07 mg x kg(-1) in the magnesium group compared with 0.29 +/- 0.01 mg x kg(-1) in the placebo group (P = 0.017). Hemodynamic variables and temperature were similar in both groups. CONCLUSION: In patients undergoing cardiac surgery, administration of magnesium sulfate, resulting in ionized levels of 1.3 mmol x L(-1), results in a 30-35 min prolongation of the neuromuscular blockade induced with intubating and maintenance doses of cisatracurium and does not alter hemodynamic stability. PMID- 12560311 TI - Increased body mass index per se is not a predictor of difficult laryngoscopy. AB - PURPOSE: We investigated the association between morbid obesity and difficult laryngoscopy (DL). METHODS: In a prospective, controlled study we evaluated the impact of different variables on the prediction of DL in 200 morbidly obese (study group-SG), and 1272 non-obese (control group-CG) patients undergoing elective surgery. Variables assessed included age, sex, body mass index (BMI), protruding, loose, and missing upper teeth, thyro-mental distance, temporo mandibular joint (TMJ) function, neck extension, and Mallampati class. A Cormack grade III or IV was considered DL. RESULTS: The SG patients were younger (P < 0.000), there were more females in the SG (P < 0.000) and more in the SG had teeth problems (P = 0.026). More patients in the SG (10% vs 1%), had obstructive sleep apnea (P < 0.001) with 90% of them in the SG having a grade III laryngoscopy. High BMI did not affect the laryngoscopy difficulty (P = 0.56). Multivariable regression analysis revealed that morbid obesity, increased age, male sex, pathology of TMJ, and higher Mallampati class, were independent predictors of DL. When interaction between the predictors and the group was added to the multivariable model, the SG was no longer a predictor by itself, rather its association with abnormal upper teeth turned to be significant for prediction of DL. CONCLUSIONS: Increased age, male sex, TMJ pathology, Mallampati 3 and 4, a history of obstructive sleep apnea and abnormal upper teeth were associated with a higher incidence of DL. The magnitude of BMI had no influence on difficulty with laryngoscopy. PMID- 12560312 TI - The ED(95) of end-tidal sevoflurane concentration for the smooth exchange of the tracheal tube for a laryngeal mask airway is 2.97%. AB - PURPOSE: Exchanging the tracheal tube for the laryngeal mask airway (the TT/LMA exchange) while the patient is still anesthetized avoids coughing associated with tracheal extubation. This study was conducted to determine the end-tidal concentration of sevoflurane that prevented response to this exchange in 50% patients (MAC(TT/LMA)). METHODS: We studied 21 adult male patients, ASA I, aged 21-54 yr. Anesthesia was induced with thiamylal and sevoflurane, and the trachea was intubated following vecuronium neuromuscular blockade. Anesthesia was maintained with sevoflurane and nitrous oxide in oxygen. After surgery, a predetermined end-tidal sevoflurane concentration was achieved and a steady state was maintained for at least 20 min. The concentration at which the TT/LMA exchange was attempted was determined by a modification of Dixon's up-and-down method with 0.25% as the step size. At the time of the TT/LMA exchange, no residual nitrous oxide > 3% was detected, and the return to normal neuromuscular function was confirmed. When the TT/LMA exchange was accomplished without coughing, movement, or airway obstruction, it was considered a smooth exchange. RESULTS: Sevoflurane MAC(TT/LMA) determined using the up-and-down method was 2.63% +/- 0.14%. The 50% effective dose obtained using a probit analysis was similar [2.53% (95% confidence limits, 2.13-2.82%)]. CONCLUSION: Sevoflurane MAC(TT/LMA) in adult male patients was 2.63% (1.54 MAC) and may be useful for the smooth exchange of the tracheal tube for the LMA in a clinical setting. PMID- 12560313 TI - Laryngeal nerve identification during thyroid surgery -- feasibility of a novel approach. AB - PURPOSE: Recurrent laryngeal nerve damage remains one of the most devastating complications of thyroid surgery. However, nerve identification is not always easy, and a reliable method to locate nerves intraoperatively is needed. METHODS: Thirty consecutive patients were anesthetized for elective thyroid surgery using a standard technique. Indications for surgery covered a broad spectrum of conditions. In the technique described, the airway is secured with a micro laryngeal tube, and a laryngeal mask airway is inserted through which a fibreoptic scope is inserted to view the larynx. Movement of the arytenoids in response to nerve stimulation can be viewed at any time on a television monitor. The airway is secure throughout the procedure and nerve identification is continuously available. RESULTS: In our study 30 patients were anesthetized and nerve stimulation used in all of them to identify both superior and recurrent laryngeal nerve. None of them developed intraoperative complications. One patient had temporary postoperative recurrent laryngeal nerve damage, which was not attributable to use of this method. CONCLUSION: On the basis of our results so far, the method described is feasible and provides a safe method of nerve location during surgery. Laryngeal nerve stimulation is likely to become an integral part of thyroid surgery. PMID- 12560314 TI - Olprinone, a phosphodiesterase III inhibitor, does not affect hypoxia-induced pial arteriolar dilatation in rabbits. AB - PURPOSE: Olprinone, a phosphodiesterase III inhibitor, is used for the treatment of heart failure or asthma. Such patients may suffer from hypoxia. However, the effect of olprinone on the cerebral vasodilator response to hypoxia remains unclear. METHODS: Rabbits were anesthetized and ventilated mechanically. The pial arteriolar diameter was determined using a cranial window and intravital microscopy. Hypoxia was induced twice in the same animal by reducing FIO(2) to 0.1. The first episode was induced during an infusion of saline, and the second during an infusion of saline (saline group; n = 8) or olprinone (1 microg x kg( 1) x min(-1), OLP1 group; n = 8 or 10 microg x kg(-1) x min(-1), OLP10 group; n = 8). The pial arteriolar responses to hypoxia were recorded and compared between the two episodes of hypoxia in each group. RESULTS: Blood gas data in the first hypoxic challenge were identical to those in the second challenge in each group. Pial arteriolar diameter increased significantly during hypoxia. In arterioles between 50-100 microm diameter, first and second hypoxia-induced pial arteriolar dilatation in OLP1 were 13 +/- 6% and 10 +/- 7% respectively (P = 0.574 ) and those in OLP10 were 16 +/- 6% and 15 +/- 7% respectively (P = 0.606). In arterioles between 25-50 microm, the results were the same as in arterioles between 50-100 microm. CONCLUSION: Olprinone does not affect the hypoxia-induced dilatation of pial arterioles in pentobarbital anesthetized rabbits. PMID- 12560315 TI - Alfentanil has no proconvulsive effect during electroconvulsive therapy. PMID- 12560316 TI - Delayed diagnosis of latex allergy. PMID- 12560317 TI - The physiologic response to laparoscopic cholecystectomy: CO(2) pneumoperitoneum vs wall lift. PMID- 12560318 TI - Repeated peripheral nerve blocks by the co-administration of ketamine, morphine, and bupivacaine attenuate trigeminal neuralgia. PMID- 12560319 TI - Evaluation of the Arrow Flex Tip Plus epidural catheter tip position and migration during continuous thoracic analgesia. PMID- 12560320 TI - Is there a need for the epiglottic bars in the laryngeal mask airway? PMID- 12560321 TI - Hyperbaric oxygen therapy (HBOT) in a child with suspected influenza-associated encephalopathy. PMID- 12560322 TI - Keilin, cytochrome, and the respiratory chain. PMID- 12560323 TI - Transforming growth factor-beta1 potentiates renal tubular epithelial cell death by a mechanism independent of Smad signaling. AB - Tubular atrophy resulting from epithelial cell loss is one of the characteristic features in the development of chronic renal interstitial fibrosis. Although the trigger(s) and mechanism for tubular cell loss remain undefined, the hyperactive transforming growth factor (TGF)-beta1 signaling has long been suspected to play an active role. Here we demonstrate that although TGF-beta1 did not induce cell death per se, it dramatically potentiated renal tubular cell apoptosis initiated by other death cues in vitro. Pre-incubation of human kidney epithelial cells (HKC) with TGF-beta1 markedly promoted staurosporine-induced cell death in a time and dose-dependent manner. TGF-beta1 dramatically accelerated the cleavage and activation of pro-caspase-9, but not pro-caspase-8, in HKC cells. This event was followed by an accelerated activation of pro-caspase-3. To elucidate the mechanism underlying TGF-beta1 promotion of tubular cell death, we investigated the signaling pathways activated by TGF-beta1. Both Smad-2 and p38 mitogen activated protein (MAP) kinase were rapidly activated by TGF-beta1, as demonstrated by the early induction of phosphorylated Smad-2 and p38 MAP kinase, respectively. We found that overexpression of inhibitory Smad-7 completely abolished Smad-2 phosphorylation and activation induced by TGF-beta1 but did not inhibit TGF-beta1-induced apoptosis. However, suppression of p38 MAP kinase with chemical inhibitor SC68376 not only abolished p38 MAP kinase phosphorylation but also obliterated apoptosis induced by TGF-beta1. These results suggest that hyperactive TGF-beta1 signaling potentiates renal tubular epithelial cell apoptosis by a Smad-independent, p38 MAP kinase-dependent mechanism. PMID- 12560324 TI - Ecto 5'-nucleotidase and nonspecific alkaline phosphatase. Two AMP-hydrolyzing ectoenzymes with distinct roles in human airways. AB - In human airways, extracellular adenosine regulates epithelial functions supporting mucociliary clearance, an important airway defense mechanism against bacterial infection. Thus, defining the mechanisms of adenosine generation is critical for elucidating the role of this nucleoside in airway homeostasis. In this study, we identified the source of adenosine on the mucosal surface of human airway epithelia. Polarized primary cultures of human nasal or bronchial epithelial cells were assayed for transepithelial transport, cytosolic and cell surface adenosine production. Ussing chamber experiments indicated that serosal 1 microM [(3)H]adenosine was not transported to the mucosal compartment. Messenger RNA for the cytosolic AMP-specific 5'-nucleotidase (CN-I) was not detected in human bronchial epithelial cells, suggesting that mucosal adenosine did not originate from intracellular pools. In contrast, extracellular 0.1 mm ATP was rapidly dephosphorylated into adenosine on the mucosal epithelial surface. We identified two ectonucleotidases that mediated the conversion of AMP to adenosine: ecto 5'-nucleotidase (ecto 5'-NT, CD73) and alkaline phosphatase (AP). Both mucosal and serosal epithelial surfaces displayed ecto 5'-NT activity (K(m) = 14 microM, V(max) = 0.5 nmol x min(-1) x cm(-2)), whereas AP activity was restricted to the mucosal surface (K(m,)(high) = 36 microM, V(max) = 1.2 nmol x min(-1) x cm(-2); K(m,)(low) = 717 microM, V(max) = 2.8 nmol x min(-1) x cm(-2)). In bronchial cultures and tissues, ecto 5'-NT accounted for >80% of total activity toward 0.01 mm AMP, compared with <15% for 5 mm AMP. The proximal airway AP isoform was identified as nonspecific AP (NS AP) by levamisole sensitivity and mRNA expression. The two ectoenzymes presented opposite airway distributions, ecto 5'-NT and NS AP mRNA dominating in higher and lower airways, respectively. Collectively, these experiments support a major role for extracellular nucleotide catalysis and for ecto 5'-NT and NS AP in the regulation of adenosine concentrations on airway surfaces. PMID- 12560325 TI - Glucocorticoids repress transcription of phosphoenolpyruvate carboxykinase (GTP) gene in adipocytes by inhibiting its C/EBP-mediated activation. AB - The cytosolic form of the phosphoenolpyruvate carboxykinase (PEPCK-C) gene is selectively expressed in several tissues, primarily in the liver, kidney, and adipose tissue. The transcription of the gene is reciprocally regulated by glucocorticoids in these tissues. It is induced in the liver and kidney but repressed in the white adipose tissue. To elucidate which adipocyte-specific transcription factors participate in the repression of the gene, DNase I footprinting analyses of nuclear proteins from 3T3-F442A adipocytes and transient transfection experiments in NIH3T3 cells were utilized. Glucocorticoid treatment slightly reduced the nuclear C/EBP alpha concentration but prominently diminished the binding of adipocyte-derived nuclear proteins to CCAAT/enhancer-binding protein (C/EBP) recognition sites, without affecting the binding to nuclear receptor sites in the PEPCK-C gene promoter. Of members of the C/EBP family of transcription factors, C/EBP alpha was the strongest trans-activator of the PEPCK C gene promoter in the NIH3T3 cell line. The glucocorticoid receptor (GR), in the presence of its hormone ligand, inhibited the activation of the PEPCK-C gene promoter by C/EBP alpha or C/EBP beta but not by the adipocyte-specific peroxisome proliferator-activated receptor gamma 2. This inhibition effect was similar using the wild type or mutant GR and did not depend on GR binding to the DNA. The glucocorticoid response unit (GRU) in the PEPCK-C gene promoter (-2000 to +73) restrained C/EBP alpha-mediated trans-activation, because mutation of each single GRU element increased this activation by 3-4-fold. This series of GRU mutations were repressed by wild type GR to the same percent as was the nonmutated PEPCK-C gene promoter. In contrast, the repression by mutant GR depended on the intact AF1 site in the gene promoter, whereby mutation of the AF1 element abolished the repression. PMID- 12560326 TI - Photoaffinity labeling with a neuroactive steroid analogue. 6-azi-pregnanolone labels voltage-dependent anion channel-1 in rat brain. AB - Neuroactive steroids modulate the function of gamma-aminobutyric acid, type A (GABA(A)) receptors in the central nervous system by an unknown mechanism. In this study we have used a novel neuroactive steroid analogue, 3 alpha,5 beta-6 azi-3-hydroxypregnan-20-one (6-AziP), as a photoaffinity labeling reagent to identify neuroactive steroid binding sites in rat brain. 6-AziP is an effective modulator of GABA(A) receptors as evidenced by its ability to inhibit binding of [(35)S]t-butylbicyclophosphorothionate to rat brain membranes and to potentiate GABA-elicited currents in Xenopus oocytes and human endothelial kidney 293 cells expressing GABA(A) receptor subunits (alpha(1)beta(2)gamma(2)). [(3)H]6-AziP produced time- and concentration-dependent photolabeling of protein bands of approximately 35 and 60 kDa in rat brain membranes. The 35-kDa band was half maximally labeled at a [(3)H]6-AziP concentration of 1.9 microM, whereas the 60 kDa band was labeled at higher concentrations. The photolabeled 35-kDa protein was isolated from rat brain by two-dimensional PAGE and identified as voltage dependent anion channel-1 (VDAC-1) by both matrix-assisted laser desorption ionization time-of-flight and ESI-tandem mass spectrometry. Monoclonal antibody directed against the N terminus of VDAC-1 immunoprecipitated labeled 35-kDa protein from a lysate of rat brain membranes, confirming that VDAC-1 is the species labeled by [(3)H]6-AziP. The beta(2) and beta(3) subunits of the GABA(A) receptor were co-immunoprecipitated by the VDAC-1 antibody suggesting a physical association between VDAC-1 and GABA(A) receptors in rat brain membranes. These data suggest that neuroactive steroid effects on the GABA(A) receptor may be mediated by binding to an accessory protein, VDAC-1. PMID- 12560327 TI - Efficient initiation of HIV-1 reverse transcription in vitro. Requirement for RNA sequences downstream of the primer binding site abrogated by nucleocapsid protein dependent primer-template interactions. AB - Synthesis of HIV-1 (-) strong-stop DNA is initiated following annealing of the 3' 18 nucleotides (nt) of tRNA(3)(Lys) to the primer binding site (PBS) near the 5' terminus of viral RNA. Here, we have investigated whether sequences downstream of the PBS play a role in promoting efficient (-) strong-stop DNA synthesis. Our findings demonstrate a template requirement for at least 24 bases downstream of the PBS when tRNA(3)(Lys) or an 18-nt RNA complementary to the PBS (R18), but not an 18-nt DNA primer, are used. Additional assays using 18-nt DNA-RNA chimeric primers, as well as melting studies and circular dichroism spectra of 18-nt primer:PBS duplexes, suggest that priming efficiency is correlated with duplex conformation and stability. Interestingly, in the presence of nucleocapsid protein (NC), the 24 downstream bases are dispensable for synthesis primed by tRNA(3)(Lys) but not by R18. We present data supporting the conclusion that NC promotes extended interactions between the anticodon stem and variable loop of tRNA(3)(Lys) and a sequence upstream of the A-rich loop in the template. Taken together, this study leads to new insights into the initiation of HIV-1 reverse transcription and the functional role of NC-facilitated tRNA-template interactions in this process. PMID- 12560328 TI - Identification of p21WAF1/CIP1 as a direct target of EWS-Fli1 oncogenic fusion protein. AB - Translocation t(11;22) is a karyotypic abnormality detected in over 90% of Ewing's family tumors. This translocation results in the EWS-Fli1 fusion gene, which has been shown to be a potent, single-step transforming gene. We reported previously that suppression of the EWS-Fli1 fusion protein altered the expression of G(1) regulatory cyclins and cyclin-dependent kinase inhibitors both at mRNA and protein levels, resulting in G(1) growth arrest in Ewing's family tumor cell lines. These data suggest that the G(1) regulatory molecules may be targets of the EWS-Fli1 fusion protein, which functions as an aberrant transcription factor. By using electrophoretic mobility shift assays, we show here the direct association of EWS-Fli1 fusion protein with ETS consensus sequences, which are in the promoter of the p21(WAF1/CIP1) gene. Reporter gene assays revealed that the activity of the p21(WAF1/CIP1) promoter is negatively regulated by EWS-Fli1 fusion protein through at least two ETS-binding sites in the promoter. EWS-Fli1 interacted with p300 cotransactivator and suppressed its histone acetyltransferase activity, which may explain the down-regulation of p21(WAF1/CIP1) by EWS-Fli1. In the presence of a histone deacetylase inhibitor, the histone acetyltransferase activity of the Ewing's family tumor cell was recovered resulting in the induction of p21, and the cell growth was dramatically inhibited. These results demonstrated that p21(WAF1/CIP1) might be one of the direct targets of EWS-Fli1, and that p21(WAF1/CIP1) could serve as a target for a molecularly based therapy for Ewing's family tumors. PMID- 12560329 TI - Mitochondrial membrane permeabilization and superoxide production during apoptosis. A single-cell analysis. AB - The temporal relationship between mitochondrial membrane permeabilization and reactive oxygen species production during apoptosis remains unknown. We analyzed the rate of superoxide production of human breast carcinoma cells expressing a cytochrome c-green fluorescent protein (cyt-c-GFP) fusion protein at the single cell level during apoptosis. In cells treated with the proapoptotic agents staurosporine (3 microm) or tumor necrosis factor-alpha (100 ng/ml), the release of cyt-c-GFP was individually set for each cell, and the majority of the fusion protein was released in less than 10 min. Prior to the release of the fusion protein, cells demonstrated a constant rate of superoxide production determined with the probe hydroethidine. After the release was completed, the superoxide concentration increased rapidly to a level more than 3-fold above baseline. Treatment with the broad spectrum caspase inhibitor z-Val-Ala-Asp(O-methyl) fluoromethylketone (z-VAD-fmk; 200 microm) did not alter the kinetics of the cyt c-GFP release but significantly reduced superoxide concentration after the release of cyt-c-GFP. Interestingly, treatment with z-VAD-fmk also reduced the increase in superoxide concentration in response to menadione in the absence of mitochondrial cyt-c-GFP release. Mitochondrial depolarization with the protonophore carbonyl cyanide p-trifluoromethoxy-phenylhydrazone per se did not trigger cyt-c-GFP release or an increase in superoxide production. Our data suggest that mitochondria increase their superoxide production during apoptosis directly after the quantitative release of soluble intermembrane proteins and demonstrate novel antioxidative effects of the commonly used caspase inhibitor z VAD-fmk. PMID- 12560330 TI - Suppressor of cytokine signaling-3 (SOCS-3), a potential mediator of interleukin 6-dependent insulin resistance in hepatocytes. AB - Interleukin-6 (IL-6) is one of several pro-inflammatory cytokines implicated in insulin resistance during infection, cachexia, and obesity. We recently demonstrated that IL-6 inhibits insulin signaling in hepatocytes (Senn, J. J., Klover, P. J., Nowak, I. A., and Mooney, R. A. (2002) Diabetes 51, 3391-3399). Members of the suppressors of cytokine signaling (SOCS) family associate with the insulin receptor (IR), and their ectopic expression inhibits IR signaling. Since several SOCS proteins are induced by IL-6, a working hypothesis is that IL-6 dependent insulin resistance is mediated, at least in part, by induction of SOCS protein(s) in insulin target cells. To examine the involvement of SOCS protein(s) in IL-6-dependent inhibition of insulin receptor signaling, HepG2 cells were treated with IL-6 (20 ng/ml) for periods from 1 min to 8 h. IL-6 induced SOCS-3 transcript at 30 min with a maximum effect at 1 h. SOCS-3 protein levels were also markedly elevated at 1 h. Transcript and protein levels returned to near basal levels by 2 h. SOCS-3 induction by IL-6 paralleled IL-6-dependent inhibition of IR signal transduction. Ectopically expressed SOCS-3 associated with the IR and suppressed insulin-dependent receptor autophosphorylation, insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation, association of IRS 1 with the p85 subunit of phosphatidylinositol 3-kinase, and activation of Akt. SOCS-3 was also a direct inhibitor of insulin receptor autophosphorylation in vitro. In mice exposed to IL-6 for 60-90 min, hepatic SOCS-3 expression was increased. This was associated with inhibition of hepatic insulin-dependent receptor autophosphorylation and IRS-1 tyrosine phosphorylation. These data suggest that induction of SOCS-3 in liver may be an important mechanism of IL-6 mediated insulin resistance. PMID- 12560331 TI - The unfolded protein response is required for haploid tolerance in yeast. AB - HAC1 encodes a transcription factor that mediates the unfolded protein response (UPR) in Saccharomyces cerevisiae. We characterized hac1Delta mutants in the sporulation-proficient SK1 genetic background and found a novel function for HAC1 in haploid tolerance. hac1Delta spore clones contain a diploid DNA content as determined by fluorescence-activated cell sorting and genetic analyses. Autodiploidization of hac1 spore clones occurred after germination; hac1 spores were born haploid, but efficiently generated diploid progeny during the subsequent mitotic division. Once the hac1 mutant acquired a diploid DNA content, no further ploidy increase was observed. Interestingly, the increase in genome content following meiosis was not a general property associated with hac1 spore clones; instead, it was restricted to an inability to tolerate the haploid state. Genetic analyses involving the UPR target gene KAR2 and the UPR regulator IRE1 revealed that autodiploidization associated with hac1 mutants is a consequence of its role in the UPR pathway. Inhibition of the UPR pathway induces autodiploidization, and constitutive activation of UPR target genes suppresses this response. PMID- 12560332 TI - Interaction of L2 with beta-actin directs intracellular transport of papillomavirus and infection. AB - Viruses that replicate in the nucleus, including the primary causative agent of cervical cancer, human papillomavirus type 16 (HPV16), must first cross the cytoplasm. We compared the uptake of HPV16 virus-like particles (VLPs) either with or without the minor capsid protein L2. Whereas VLPs containing only the major capsid protein L1 were diffusely distributed within the cytoplasm even 6 h post-infection, VLPs comprising both L1 and L2 exhibited a radial distribution in the cytoplasm and accumulated in the perinuclear region of BPHE-1 cells within 2 h. L2 of HPV16 or bovine papillomavirus was shown to bind to a 43-kDa cellular protein that was subsequently identified as beta-actin by matrix-assisted laser desorption ionization time-of-flight analysis. A conserved domain comprising residues 25-45 of HPV16 L2 was sufficient for interaction with beta-actin. HPV16 L2 residues 25-45 fused to green fluorescent protein, but not green fluorescent protein alone, colocalized with actin and caused cell retraction and disruption of the microfilament network. Finally, wild-type L2, but not L2 with residues 25 45 deleted, facilitated HPV16 pseudovirion infection. Thus, binding of beta-actin by L2 residues 25-45 facilitates transport of HPV16 across the cytoplasm during infection, and blockade of this novel interaction may be useful for prophylaxis. PMID- 12560333 TI - Mitochondrial ATP synthasome. Cristae-enriched membranes and a multiwell detergent screening assay yield dispersed single complexes containing the ATP synthase and carriers for Pi and ADP/ATP. AB - The terminal step of ATP synthesis in intact mitochondria is catalyzed by the ATP synthase (F(0)F(1)) that works in close synchrony with the P(i) and ADP/ATP carriers. Each carrier consists of only a single polypeptide chain in dimeric form, while the ATP synthase is highly complex consisting in animals of 17 known subunit types and more than 30 total subunits. Although structures at high resolution have been obtained for the water-soluble F(1) part of the ATP synthase consisting of only five subunit types, such structures have not been obtained for either the complete ATP synthase or the P(i) and ADP/ATP carriers. Here, we report that all three proteins are localized in highly purified cristae-like vesicles obtained by extensive subfractionation of the mitochondrial inner membrane. Moreover, using a multiwell detergent screening assay, 4 nonionic detergents out of 80 tested were found to disperse these cristae-like vesicles into single soluble complexes or "ATP synthasomes" that contain the ATP synthase in association with the P(i) and ADP/ATP carriers. These studies offer new mechanistic insights into the terminal steps of oxidative phosphorylation in mitochondria and set the stage for future structural efforts designed to visualize in atomic detail the entire complex involved. They also provide evidence that the cristae are a subcompartment of the inner membrane. PMID- 12560334 TI - A functional domain of the alpha1 subunit of soluble guanylyl cyclase is necessary for activation of the enzyme by nitric oxide and YC-1 but is not involved in heme binding. AB - Soluble guanylyl cyclase is a heterodimeric enzyme consisting of an alpha(1) and a beta(1) subunit and is an important target for endogenous nitric oxide and the guanylyl cyclase modulator YC-1. The activation of the enzyme by both substances is dependent on the presence of a prosthetic heme group. It has been unclear whether this prosthetic heme group is sandwiched between the alpha(1) and beta(1) subunits or whether it exclusively binds to the beta(1) subunit. Here we analyze progressive amino-terminal deletion mutants of the human alpha(1) subunit after co-expression with the human beta(1) subunit in the baculovirus/Sf9 system. Spectral, biochemical, and pharmacological analysis shows that the first 259 amino acids of the alpha(1) subunit can be deleted without loss of sensitivity to nitric oxide (NO) or YC-1 or loss of heme binding of the respective enzyme complex with the beta(1) subunit. This is in contrast to previous data indicating that NO sensitivity and a functional heme binding site requires full-length amino termini of bovine alpha(1) and beta(1) subunits. Further deletion of the first 364 amino acids of the alpha(1) subunit leads to an enzyme complex with preserved heme binding but loss of sensitivity to NO or YC-1 despite induction of the typical spectral shift by NO binding to the prosthetic heme group. We conclude that 1) the amino-terminal part of the alpha(1) subunit is not involved in heme binding and 2) amino acids 259-364 of the alpha(1) subunit represent an important functional domain for the transduction of the NO activation signal and likely represent the target for NO-sensitizing substances like YC-1. PMID- 12560335 TI - Activation of RGS9-1GTPase acceleration by its membrane anchor, R9AP. AB - The GTPase-accelerating protein (GAP) complex RGS9-1.G beta(5) plays an important role in the kinetics of light responses by accelerating the GTP hydrolysis of G alpha(t) in vertebrate photoreceptors. Much, but not all, of this complex is tethered to disk membranes by the transmembrane protein R9AP. To determine the effect of the R9AP membrane complex on GAP activity, we purified recombinant R9AP and reconstituted it into lipid vesicles along with the photon receptor rhodopsin. Full-length RGS9-1.G beta(5) bound to R9AP-containing vesicles with high affinity (K(d) < 10 nm), but constructs lacking the DEP (dishevelled/EGL 10/pleckstrin) domain bound with much lower affinity, and binding of those lacking the entire N-terminal domain (i.e. the dishevelled/EGL-10/pleckstrin domain plus intervening domain) was not detectable. Formation of the membrane bound complex with R9AP increased RGS9-1 GAP activity by a factor of 4. Vesicle titrations revealed that on the time scale of phototransduction, the entire reaction sequence from GTP uptake to GAP-catalyzed hydrolysis is a membrane delimited process, and exchange of G alpha(t) between membrane surfaces is much slower than hydrolysis. Because in rod cells different pools exist of RGS9-1.G beta(5) that are either associated with R9AP or not, regulation of the association between R9AP and RGS9-1.G beta(5) represents a potential mechanism for the regulation of recovery kinetics. PMID- 12560336 TI - Characterization of endothelin-converting enzyme-2. Implication for a role in the nonclassical processing of regulatory peptides. AB - Most neuroendocrine peptides are generated by proteolysis of the precursors at basic residue cleavage sites. Prohormone convertases belonging to the subtilisin family of serine proteases are primarily responsible for processing at these "classical sites." In addition to the classical cleavages, a subset of bioactive peptides is generated by processing at "nonclassical" sites. The proteases responsible for these cleavages have not been well explored. Members of several metalloprotease families have been proposed to be involved in nonclassical processing. Among them, endothelin-converting enzyme-2 (ECE-2) is a good candidate because it exhibits a neuroendocrine distribution and an acidic pH optimum. To examine the involvement of this protease in neuropeptide processing, we purified the recombinant enzyme and characterized its catalytic activity. Purified ECE-2 efficiently processes big endothelin-1 to endothelin-1 by cleavage between Trp(21) and Val(22) at acidic pH. To characterize the substrate specificity of ECE-2, we used mass spectrometry with a panel of 42 peptides as substrates to identify the products. Only 10 of these 42 peptides were processed by ECE-2. A comparison of residues around the cleavage site revealed that ECE-2 exhibits a unique cleavage site selectivity that is related to but distinct from that of ECE-1. ECE-2 tolerates a wide range of amino acids in the P1-position and prefers aliphatic/aromatic residues in the P1'-position. However, only a small fraction of the aliphatic/aromatic amino acid-containing sites were cleaved, indicating that there are additional constraints beyond the P1- and P1' positions. The enzyme is able to generate a number of biologically active peptides from peptide intermediates, suggesting an important role for this enzyme in the biosynthesis of regulatory peptides. Also, ECE-2 processes proenkephalin derived bovine adrenal medulla peptides, and this processing leads to peptide products known to have differential receptor selectivity. Finally, ECE-2 processes PEN-LEN, an endogenous inhibitor of prohormone convertase 1, into products that do not inhibit the enzyme. Taken together, these results are consistent with an important role for ECE-2 in the processing of regulatory peptides at nonclassical sites. PMID- 12560337 TI - Mechanism of endothelial cell NADPH oxidase activation by angiotensin II. Role of the p47phox subunit. AB - Endothelial cells express a constitutively active phagocyte-type NADPH oxidase whose activity is augmented by agonists such as angiotensin II. We recently reported (Li, J.-M., and Shah, A. M. (2002) J. Biol. Chem. 277, 19952-19960) that in contrast to neutrophils a substantial proportion of the NADPH oxidase in unstimulated endothelial cells exists as preassembled intracellular complexes. Here, we investigate the mechanism of angiotensin II-induced endothelial NADPH oxidase activation. Angiotensin II (100 nmol/liter)-induced reactive oxygen species production (as measured by dichlorohydrofluorescein fluorescence or lucigenin chemiluminescence) was completely absent in coronary microvascular endothelial cells isolated from p47(phox) knockout mice. Transfection of p47(phox) cDNA into p47(phox-/-) cells restored the angiotensin II response, whereas transfection of antisense p47(phox) cDNA into wild-type cells depleted p47(phox) and inhibited the angiotensin II response. In unstimulated human microvascular endothelial cells, there was significant p47(phox)-p22(phox) complex formation but minimal detectable p47(phox) phosphorylation. Angiotensin II induced rapid serine phosphorylation of p47(phox) (within 1 min, peaking at approximately 15 min), a 1.9 +/- 0.1-fold increase in p47(phox)-p22(phox) complex formation and a 1.6 +/- 0.2-fold increase in NADPH-dependent O(2)-* production (p < 0.05). p47(phox) was redistributed to "nuclear" and membrane-enriched cell fractions. These data indicate that angiotensin II-stimulated endothelial NADPH oxidase activity is regulated through serine phosphorylation of p47(phox) and its enhanced binding to p22(phox). PMID- 12560338 TI - Glucosylceramide, a neutral glycosphingolipid anticoagulant cofactor, enhances the interaction of human- and bovine-activated protein C with negatively charged phospholipid vesicles. AB - The effect of glucosylceramide (GlcCer) on activated protein C (APC)-phospholipid interactions was examined using fluorescence resonance energy transfer. Human APC, labeled with either fluorescein (Fl-APC) or dansyl (DEGR-APC) donor, bound to phosphatidylcholine/phosphatidylserine (PC/PS, 9:1 w/w) vesicles containing octadecylrhodamine (OR) acceptor with a K(d) (app) = 16 micro g/ml, whereas Fl APC (or DEGR-APC) bound to PC/PS/GlcCer(OR) (8:1:1) vesicles with a K(d) (app) = 3 micro g/ml. This 5-fold increase in apparent affinity was not species-specific since bovine DEGR-APC also showed a similar GlcCer-dependent enhancement of binding of APC to PC/PS vesicles. From the efficiency of fluorescence resonance energy transfer, distances of closest approach of approximately 63 and approximately 64 A were estimated between the dansyl on DEGR-APC and rhodamine in PC/PS/GlcCer(OR) and PC/PS(OR), respectively, assuming kappa(2) = 2/3. DEGR-APC bound to short chain C8-GlcCer with an apparent K(d) of 460 nm. The presence of C8-GlcCer selectively enhanced the binding of C16,6-NBD-phosphatidylserine but not C16,6-7-nitrobenz-2-oxa-1,3-diazole (NBD)-phosphatidylcholine to coumarin labeled APC. These data suggest that APC binds to GlcCer, that PC/PS/GlcCer vesicles like PC/PS vesicles bind to the N-terminal gamma-carboxyglutamic acid domain of APC, and that one mechanism by which GlcCer enhances the activity of APC is by increasing its affinity for membrane surfaces containing negatively charged phospholipids. PMID- 12560339 TI - Localization of p21-activated protein kinase gamma-PAK/Pak2 in the endoplasmic reticulum is required for induction of cytostasis. AB - The intracellular localization and physiological functions of the p21-activated protein kinase gamma-PAK have been examined in human embryonic kidney 293T and COS-7 cells. At 1-4 days post-transfection, cell division is inhibited by the expression of wild type (WT) gamma-PAK and the mutant S490A, whereas cells expressing S490D and the inactive mutants K278R and T402A grow exponentially, indicating a role for gamma-PAK in the induction of cytostasis. WT gamma-PAK and S490A are localized in a region surrounding the nucleus identified as the endoplasmic reticulum (ER), as determined by immunofluorescence, whereas K278R, T402A, and S490D lack localization. As shown by sucrose density gradient centrifugation, WT gamma-PAK, S490A, and endogenous gamma-PAK are distributed among the high density (ER-associated), intermediate density, and low density fractions, whereas the mutants that do not inhibit cell division are present only as soluble enzyme. The amount of endogenous gamma-PAK associated with the particulate fractions is increased 4-fold when cell division is inhibited by ionizing radiation. gamma-PAK in the ER and intermediate density fractions has high specific activity and is active, whereas the soluble form of gamma-PAK has low activity and is activable. The importance of localization of gamma-PAK is supported by data with the C-terminal mutants S490D and Delta 488; these mutants have high levels of protein kinase activity but do not induce cytostasis and are not bound to the ER. A model for the induction of cytostasis by gamma-PAK through targeting of gamma-PAK to the ER is presented in which gamma-PAK activity and Ser 490 are implicated in the regulation of cytostasis. PMID- 12560340 TI - The Roles of N- and C-terminal determinants in the activation of the Kv2.1 potassium channel. AB - The human and rat forms of the Kv2.1 channel have identical amino acids over the membrane-spanning regions and differ only in the N- and C-terminal intracellular regions. Rat Kv2.1 activates much faster than human Kv2.1. Here we have studied the role of the N- and C-terminal residues that determine this difference in activation kinetics between the two channels. For this, we constructed mutants and chimeras between the two channels, expressed them in oocytes, and recorded currents by two-electrode voltage clamping. In the N-terminal region, mutation Q67E in the rat channel displayed a slowing of activation relative to rat wild type, whereas mutation D75E in the human channel showed faster activation than human wild type. In the C-terminal region, we found that some residues within the region of amino acids 740-853 ("CTA" domain) were also involved in determining activation kinetics. The electrophysiological data also suggested interactions between the N and C termini. Such an interaction was confirmed directly by using a glutathione S-transferase (GST) fusion protein with the N terminus of Kv2.1, which we showed to bind to the C terminus of Kv2.1. Taken together, these data suggest that exposed residues in the T1 domain of the N terminus, as well as the CTA domain in the C terminus, are important in determining channel activation kinetics and that these N- and C-terminal regions interact. PMID- 12560341 TI - Nuclear localization of the cell cycle regulator CDH1 and its regulation by phosphorylation. AB - The anaphase-promoting complex activated by CDC20 and CDH1 is a major ubiquitination system that controls the destruction of cell cycle regulators. Exactly how ubiquitination is regulated in time and space is incompletely understood. Here we report on the cell cycle-dependent localization of CDH1 and its regulation by phosphorylation. CDH1 localizes dynamically to the nucleus during interphase and to the centrosome during metaphase and anaphase. The nuclear accumulation of CDH1 correlates with a reduction in the steady-state amount of cyclin A, but not of cyclin E. A nuclear localization signal conserved in various species was identified in CDH1, and it sufficiently targets green fluorescent protein to the nucleus. Interestingly, a CDH1-4D mutant mimicking the hyperphosphorylated form was constitutively found in the cytoplasm. In further support of the notion that phosphorylation inhibits nuclear import, the nuclear localization signal of CDH1 with two phospho-accepting serine/threonine residues changed into aspartates was unable to drive heterologous protein into the nucleus. On the other hand, abolition of the cyclin-binding ability of CDH1 has no influence on its nuclear localization. Taken together, our findings document the phosphorylation-dependent localization of CDH1 in vertebrate cells. PMID- 12560342 TI - Multiplex minisequencing screen for common Southeast Asian and Indian beta thalassemia mutations. AB - BACKGROUND: Beta-thalassemia is endemic to many regions in Southeast Asia and India, and <20 beta-globin gene mutations account for > or =90% of beta thalassemia alleles in these places. We describe a multiplex minisequencing assay to detect these common mutations. METHODS: Gap-PCR was used to simultaneously amplify the beta-globin gene from genomic DNA and to detect the Delta619bp deletion mutation. Multiplex minisequencing was then performed on the amplified beta-globin fragment to detect an additional 15 common Southeast Asian and Indian beta-thalassemia mutations. Site-specific primers of different lengths were subjected to multiple rounds of annealing and single-nucleotide extension in the presence of thermostable DNA polymerase and the four dideoxynucleotides, each labeled with a different fluorophore. Minisequencing products were separated and detected by capillary electrophoresis, followed by automated genotyping. The optimized assay was subjected to a double-blind validation analysis of 89 beta thalassemia and wild-type DNA samples of known genotype. RESULTS: Homozygous wild type or mutant DNA samples produced electropherograms containing only a single colored peak for each mutation site, whereas samples heterozygous for a specific mutation displayed two different-colored peaks for that mutation site. Samples were automatically genotyped based on color and position of primer peaks in the electropherogram. In the double-blind validation analysis, all 89 DNA samples were genotyped correctly (100% assay specificity). CONCLUSIONS: The described semiautomated multiplex minisequencing assay can detect the most common Southeast Asian and Indian beta-thalassemia mutations, is amenable to high-throughput scale up, and may bring population-based screening of beta-thalassemia in endemic regions a step closer to implementation. PMID- 12560343 TI - HER-2/neu gene copy number quantified by real-time PCR: comparison of gene amplification, heterozygosity, and immunohistochemical status in breast cancer tissue. AB - BACKGROUND: Amplification of the oncogene HER-2/neu influences breast cancer pathogenesis, and therapy and prognosis may be affected by the degree of amplification. The extent of amplification or protein overexpression typically is analyzed by fluorescence in situ hybridization or immunohistochemistry (IHC), but quantitative PCR techniques have been described that may provide alternatives to these methods. METHODS: We developed a rapid-cycle, real-time PCR assay for quantification of HER-2/neu gene status. We compared results obtained with this assay with short tandem repeat findings by capillary electrophoresis (CE) and with protein overexpression assessments by IHC. Accuracy and linearity were tested on cell lines and with simulation experiments. We analyzed the amplification of HER-2/neu in 51 clinical tissue samples from patients with suspected breast cancer. RESULTS: The intra- and interrun CVs for HER-2/neu quantification by real-time PCR were 12% and 18%, and the CV for different simulated amplification and deletion experiments was <7%. The results for HER 2/neu gene status in cell lines matched the values reported in literature. We detected HER-2/neu amplification by real-time PCR in 11 samples, all from patients with invasive ductal carcinoma. Allelic imbalances were found by CE analyses in three samples and by protein overexpression in six samples; five of these were also detected by real-time PCR. Comparison of the quantification results with known prognostic indices yielded results similar to those reported in several other published studies. CONCLUSIONS: The assay is suitable for accurate and precise quantification of HER-2/neu copy numbers in tumor tissue samples obtained in routine clinical practice. PMID- 12560344 TI - Real-time reverse transcription-PCR expression profiling of the complete human ATP-binding cassette transporter superfamily in various tissues. AB - BACKGROUND: ATP-binding cassette (ABC) transporters are involved in many physiologic processes, such as lipid transport, sterol homeostasis, immune mechanisms, and drug transport, and cause various human inherited diseases. Thus, the analysis of ABC transporter mRNA expression profiles for basic research, especially in the field of lipid metabolism, for clinical diagnosis, and for monitoring of drug effects is of great interest. METHODS: We have developed a rapid, accurate, and highly sensitive real-time reverse transcription-PCR (RT PCR) method for detection and quantification of all 47 currently known members of the ABC transporter superfamily. Our expression analysis is based on relative quantification using a calibration curve method. With our assay, expression monitoring of a large number of RNA samples in a 384-well format with only 50 ng of total RNA is possible. RESULTS: In contrast to previous expression analyses of single ABC genes, our method allows the rapid and complete analysis of all ABC transporters in given RNA samples. We used our newly established expression panel to study the gene expression of all human ABC transporters in 20 different human tissues. As a result, we identified tissues with high transcriptional activity for ABC transporters. These organs are mainly involved in secretory function (adrenal gland), metabolic function (liver), barrier function (lung, trachea, small intestine), and tropic function (placenta, uterus). CONCLUSIONS: Our RT-PCR assay allows rapid, high-throughput transcriptional profiling of the complete ABC transporter superfamily and thus provides a new enabling tool for research, clinical diagnosis of disease, and drug testing and development. PMID- 12560345 TI - Evaluation of cell-free fetal DNA as a second-trimester maternal serum marker of Down syndrome pregnancy. AB - BACKGROUND: Second-trimester cell-free fetal DNA (studied only in pregnancies with male fetuses) is higher in maternal serum samples from women carrying Down syndrome fetuses than in unaffected pregnancies. In this study we evaluated the potential performance of fetal DNA as a screening marker for Down syndrome. METHODS: Data on maternal serum fetal DNA concentrations and the corresponding concentrations of the quadruple serum markers were available from 15 Down syndrome cases, each matched for gestational age and length of freezer storage, with 5 control samples. Analyte values were expressed as multiple(s) of the control or population median. Screening performance of fetal DNA, both alone and when added to estimates of quadruple marker performance, was determined after modeling using univariate and multivariate gaussian distribution analysis. RESULTS: The median fetal DNA concentration in Down syndrome cases was 1.7 times higher than in controls. In univariate analysis, fetal DNA gave a 21% detection rate at a 5% false-positive rate. When added to quadruple marker screening, fetal DNA increased the estimated detection rate from 81% to 86% at a 5% false-positive rate. CONCLUSIONS: Cell-free fetal DNA, measured in maternal serum, can modestly increase screening performance above what is currently available in the second trimester. If and when maternal serum fetal DNA can be measured in pregnancies with both male and female fetuses, the utility and cost-effectiveness of adding it as a Down syndrome screening marker should be assessed. PMID- 12560346 TI - Comparison of two assays for human kallikrein 2. AB - BACKGROUND: We compared two recently developed research assays for the measurement of human kallikrein 2 (hK2) in serum: one fully automated assay (Beckman Coulter Access immunoanalyzer) and one manual assay based on the DELFIA technology. METHODS: We used two subsets of clinical specimens consisting of 48 samples from prostate cancer patients and 210 samples from participants in an ongoing screening study (ERSPC). Both subsets were measured in the Rotterdam laboratory, and the prostate cancer samples were used for analytical comparison with the originating sites for the assays: Beckman Coulter Research Department (San Diego, CA) and Turku University (Turku, Finland). RESULTS: Both the Beckman Coulter and the Turku assays performed very similarly between the Rotterdam laboratory and the originating sites: the R(2) value for both comparisons was 0.99, and the slope difference between sites was <20%. Deming regression analysis of the DELFIA (y) and Access (x) assays yielded the following: for the prostate cancer group, y = 1.17x - 0.01 (R(2) = 0.88; n = 48); and for the ERSPC group, y = 0.62x - 0.01 (R(2) = 0.77). Breakdown of the latter group into subgroups (nondiseased, benign prostatic hyperplasia, and prostate cancer samples) gave only minor differences. The Access calibrators were underrecovered by 13% in the DELFIA assay, whereas the DELFIA calibrators were overrecovered by 45% in the Access assay. CONCLUSION: The DELFIA and Access assays for hK2, which have similar analytical features, show differences that cannot be explained by calibration. PMID- 12560347 TI - Minor increases in plasma troponin I predict decreased left ventricular ejection fraction after high-dose chemotherapy. AB - BACKGROUND: Increased cardiac troponin I (cTnI) in patients treated with high dose chemotherapy (HDCT) for aggressive malignancy has been proposed as an early marker of late HDCT-induced cardiac dysfunction. We investigated whether cTnI measured by the Stratus CS (Dade Behring) would allow detection of minimal cTnI increases in patients treated with HDCT. METHODS: Plasma cTnI concentrations were determined in 179 consecutive patients before HDCT, at the end of the treatment, and after 12, 24, 36, and 72 h. Cardiac function was explored by echocardiography, and left ventricular ejection fraction (LVEF) was recorded during follow-up. The greatest variation in LVEF from the baseline value was used as a measure of cardiac damage. RESULTS: In 99 healthy volunteers, the 99th percentile was at 0.07 microg/L. On the basis of ROC curve analysis (area under the curve, 0.89), a cutoff of 0.08 microg/L was chosen (sensitivity, 82%; specificity, 77%). cTnI > or =0.08 microg/L occurred in 57 patients (32%) with echocardiographic monitoring revealing a mean decrease in LVEF of 18%. In comparison, the group of cTnI-negative patients had a mean decrease in LVEF of 2.5% (P <0.001). CONCLUSIONS: Plasma cTnI, as measured with the Stratus CS, can detect minor myocardial injury in patients treated with HDCT. PMID- 12560348 TI - Benign prostate-specific antigen (BPSA) in serum is increased in benign prostate disease. AB - BACKGROUND: BPSA is a "benign" form of free prostate-specific antigen (PSA) that is increased in prostate transition zone tissues of men with pathologic benign prostatic hyperplasia (BPH). We developed an immunoassay to determine the concentration of BPSA in the serum of men with BPH. METHODS: The BPSA antigen was purified by HPLC, and murine monoclonal antibodies were prepared by standard methods. A fluorogenic ELISA was developed with high specificity for BPSA and no cross-reactivity with other forms of PSA. RESULTS: The BPSA immunoassay had a lower limit of detection of 6 ng/L and a cross-reactivity of <1% with all other clipped and nonclipped forms of PSA. The BPSA antibody was specific for the internal Lys(182) cleavage site that characterizes BPSA. Biopsy-negative men with a median total PSA of 4.8 micro g/L had a median of 0.22 micro g/L BPSA, representing 25% of the free PSA in serum. BPSA ranged from 0% to 60% of the free PSA in serum. BPSA in a cohort of cancer serum also comprised 25% of the free PSA. Control serum from women or men without increased PSA had nondetectable BPSA. CONCLUSIONS: BPSA is a significant percentage of the free PSA in BPH serum but not in control serum. The presence of prostate cancer does not alter the relative proportions of BPSA in sera with <10 micro g/L PSA. BPSA has a wide distribution of concentrations in the serum and may provide clinical information for the study of men with BPH. PMID- 12560349 TI - Differences in nucleotide hydrolysis contribute to the differences between erythrocyte 6-thioguanine nucleotide concentrations determined by two widely used methods. AB - BACKGROUND: Measurement of 6-thioguanine nucleotide (6-TGN) concentrations in erythrocytes is widely accepted for use in optimization of thiopurine therapy. Various chromatographic methods have been developed for this purpose. In preliminary experiments we observed a considerable difference between 6-TGN concentrations determined with two widely used methods published by Lennard (Lennard L. J Chromatogr 1987;423:169-78) and by Dervieux and Boulieu (Dervieux T, Boulieu R. Clin Chem 1998;44:551-5). We therefore investigated methodologic differences between the two procedures with respect to hydrolysis of 6-TGNs to 6 thioguanine (6-TG) in more detail. METHODS: We analyzed 6-TGNs in erythrocyte preparations (n = 50) from patients on azathioprine therapy by both methods, using the original protocols. In one set of experiments, we replaced the 0.5 mol/L sulfuric acid in the Lennard method with the 1 mol/L perchloric acid used by Dervieux and Boulieu. In a second set of experiments, we investigated the effect of various dithiothreitol (DTT) concentrations on 6-TG recovery with both methods. In a third set of experiments, we determined the effect of hydrolysis time on both protocols. RESULTS: Direct comparison of both methods showed that 6 TGN concentrations were, on average, 2.6-fold higher in the Dervieux-Boulieu method over the concentration range tested, although the correlation (r = 0.99; P <0.001) was good. Replacement of sulfuric acid by perchloric acid reduced this difference to approximately 1.4-fold (r = 0.99; P <0.001). Increasing the DTT concentration enhanced 6-TG recovery. The hydrolysis time used in the Lennard method (1 h) was not sufficient to achieve complete hydrolysis. CONCLUSIONS: The difference between 6-TGN concentrations measured by the two methods is attributable, at least in part, to differences in the extent of nucleotide hydrolysis. For optimization of thiopurine therapy, method-dependent therapeutic ranges are necessary, which precludes comparison of results from clinical studies derived with these methods. Efforts must therefore be made to standardize the analytical procedures for the determination of 6-TGN. PMID- 12560350 TI - New enzymatic assay for glycohemoglobin. AB - BACKGROUND: Previous methods to measure glycohemoglobin (GHb) have been time consuming or imprecise; we therefore developed a new enzymatic assay for GHb. METHODS: Blood cells were first hemolyzed, and hemoglobin was digested with protease to yield fructosyl amino acid. Fructosyl amino acid oxidase acts on the fructosyl amino acid and generates hydrogen peroxide, which reacts with chromogens in the presence of peroxidase. Total hemoglobin was measured spectrometrically in the same reaction tube. The results were reported as the ratio of the concentrations of GHb and hemoglobin. RESULTS: The measured values were comparable to those determined with a HPLC method and with an immunoassay in blood samples from 2854 patients with diabetes. Regression analysis for the enzymatic assay (y) vs the HPLC method (x) produced the following: r = 0.979; slope, 0.994 [95% confidence interval (CI), 0.986-1.001]; y-intercept, 0.04% (95% CI, -0.09% to 0.01%); n = 2854. For the enzymatic assay (y) vs the immunoassay (x), the regression statistics were as follows: r = 0.982; slope, 1.002 (95% CI, 0.995-1.009); y-intercept, 0% (95% CI, -0.05% to 0.05%); n = 2854. CONCLUSIONS: The values measured by the new enzymatic assay are sufficiently correlated with those of the conventional HPLC method and immunoassay, but the proposed assay for GHb is rapid and has high precision. PMID- 12560351 TI - Detection of anti-human T-lymphotropic virus type I antibody in whole blood by a novel counting immunoassay. AB - BACKGROUND: Assays to screen for and confirm the presence of the antibody for human T-lymphotropic virus type I (HTLV-I) are currently performed with serum or plasma. We developed and evaluated a new counting immunoassay (CIA) for the detection of HTLV-I antibody in whole blood, using recombinant and synthetic peptide antigens. METHODS: We assessed the CIA for detection of HTLV-I antibody in whole blood and plasma. The CIA is an immunity-measuring method that combines latex agglutination with particle-counting technology. The numbers of agglutinated latex particles, single latex particles, and blood cells in a sample are measured based on differences in particle size between latex particles and blood cells. RESULTS: The CIA and ELISA methods were in agreement for all 24 plasma samples tested, including those from 6 patients with HTLV-I-associated diseases, 6 HTLV-I carriers, and 12 HTLV-I antibody-negative individuals. The concordance between the ELISA (plasma) and the CIA (whole blood) for samples from 24 patients was 100%. The concordance between a particle agglutination method (plasma) and the CIA (plasma or whole blood) for 1065 patients was 99.5%. The concordance between results obtained for 1065 pairs of plasma and whole blood samples with the CIA method was 100%. HTLV-I antibody in whole blood was stable for 3 days after blood collection. With this CIA method, results were available within 15 min. CONCLUSIONS: The CIA method can be used in screening for HTLV-I. The use of whole blood rather than serum or plasma reduces the sample volume and number of blood collections required, as well as assay time. PMID- 12560352 TI - Influence of urinary stones on the composition of a 24-hour urine sample. AB - BACKGROUND: It can be assumed that stones in the urinary tract continuously increase in size by incorporating material from urine. Consequently, urine will exhibit depleted concentrations of lithogenic constituents when urinary stones are present in the patient's urinary tract. METHODS: To calculate the influence of the depletion effect, we considered two different models of stone growth. In the first model, the increase in stone size depends only on the urinary concentration of a lithogenic substance; the second model also considers the surface area of the growing stone. The case of only one kidney being affected by stone formation is considered separately. We discuss example calculations involving the formation of calcium oxalate. RESULTS: The calculated depletion effects are of a nonnegligible order of magnitude. Assuming both a measured oxalate concentration of, e.g., 0.37 mmol/L and a reasonable in vivo stone growing rate of 10 mm(3)/day, a relative underestimation of the real "in situ" oxalate concentration between approximately 21% (model 1) and approximately 42% (model 2) occurs. The depletion effect increases markedly with increasing stone growth rate. CONCLUSIONS: Metabolic status can be evaluated correctly only in patients who have been declared "stone-free", e.g., after stone removal. Because the expected stone-related depletion effect in most cases is of high clinical relevance, we recommend estimating the effect of the order of magnitude of the depletion on actual urinary composition. PMID- 12560353 TI - Determination of choline, betaine, and dimethylglycine in plasma by a high throughput method based on normal-phase chromatography-tandem mass spectrometry. AB - BACKGROUND: The quaternary ammonium compounds, choline and betaine, and dimethylglycine (DMG) reside along a metabolic pathway linked to the synthesis of neurotransmitters and membrane phospholipids and to homocysteine remethylation and, therefore, folate status. Lack of a convenient, high-throughput method for the determination of these compounds has prevented population-based studies of their possible associations with lifestyle, nutrition, and chronic diseases. METHODS: Serum or plasma samples were deproteinized by mixing with three volumes of acetonitrile that contained d(9)-choline and d(9)-betaine as internal standards. We used a normal-phase silica column for the separation of choline (retention time, 2.8 min), betaine (1.3 min), DMG (1.15 min), and internal standards, which were detected as positive ions by tandem mass spectroscopy in the multiple-reaction monitoring mode, using the molecular transitions m/z 104- >60 (choline), m/z 113-->69 (d(9)-choline), m/z 118-->59 (betaine), m/z 127-->68 (d(9)-betaine), and m/z 104-->58 (DMG). RESULTS: For all three metabolites, the assay was linear in the range 0.4-400 micromol/L, and the lower limit of the detection (signal-to-noise ratio = 5) was < or =0.3 micromol/L. The within- and between-day imprecision (CVs) was 2.1-7.2% and 3.5-8.8%, respectively. The analytical recovery was 87-105%. The fasting plasma concentrations (median, 25th 75th percentiles) were 8.0 (7.0-9.3) micromol/L for choline, 31.7 (27.0-41.1) micromol/L for betaine, and 1.66 (1.30-2.02) micromol/L for DMG in 60 healthy blood donors. In individuals who had eaten a light breakfast, plasma concentrations of all three metabolites were significantly (25-30%) higher than in fasting individuals. CONCLUSION: This is the first method for the combined measurement of choline, betaine, and DMG in human plasma or serum. The assay is characterized by simple sample preparation, no derivatization, high throughput, imprecision (CV) <10%, detection limits below the values seen in volunteers, and the high specificity provided by tandem mass spectroscopy. PMID- 12560354 TI - Effect of riboflavin status on the homocysteine-lowering effect of folate in relation to the MTHFR (C677T) genotype. AB - BACKGROUND: Riboflavin (vitamin B(2)) is the precursor for FAD, the cofactor for methylenetetrahydrofolate reductase (MTHFR). MTHFR catalyzes the formation of 5 methyltetrahydrofolate, which acts as a methyl donor for homocysteine remethylation. Individuals with the MTHFR 677C-->T mutation have increased plasma total homocysteine (tHcy) concentrations, particularly in association with low folate status. It has been proposed that riboflavin may act together with folate to lower plasma tHcy, particularly in individuals with the thermolabile MTHFR T variant. METHODS: We measured B-vitamin status and plasma tHcy in 126 healthy individuals 20-63 years of age (42 CC, 42 CT, and 42 TT MTHFR genotypes) at baseline and after three interventions (4 months): placebo plus natural diet; daily 400 microg folic acid supplement plus natural diet; and increased dietary folate to 400 microg/day. RESULTS: At baseline and after nutritional intervention, lower riboflavin status was associated with increased plasma tHcy concentrations. Plasma tHcy was 2.6 micromol/L higher in the lowest plasma riboflavin quartile compared with the highest (P <0.02) and was 4.2 micromol/L higher in the highest erythrocyte glutathione reductase activation coefficient (EGRAC) quartile compared with the lowest (P <0.001). This effect was not restricted to those with the T allele. Folic acid given as a 400 microg/day supplement appeared to exacerbate a tendency toward riboflavin deficiency, as suggested by an increase in the proportion of individuals with EGRAC > or =1.4 from 52% to 65% after supplementation (P <0.05). CONCLUSIONS: Folate and riboflavin interact to lower plasma tHcy, possibly by maximizing the catalytic activity of MTHFR. The effect may be unrelated to MTHFR genotype. PMID- 12560355 TI - Apolipoprotein C-III bound to apolipoprotein B-containing lipoproteins in obese girls. PMID- 12560356 TI - Broad-range ribosomal RNA real-time PCR after removal of DNA from reagents: melting profiles for clinically important bacteria. PMID- 12560357 TI - Detection of microchimeric cells in the peripheral blood of nonpregnant women is enhanced by magnetic cell sorting before PCR. PMID- 12560358 TI - Real-time reverse transcription-PCR detects KS1/4 mRNA in mediastinal lymph nodes from patients with non-small cell lung cancer. PMID- 12560359 TI - Reference intervals for plasma homocysteine by the AxSYM immunoassay after collection in fluoride tubes. PMID- 12560360 TI - Characterization of biochemical and clinical correlates of hypocholesterolemia after hepatectomy. PMID- 12560361 TI - Deletion of the C4-CYP21 repeat module leading to the formation of a chimeric CYP21P/CYP21 gene in a 9.3-kb fragment as a cause of steroid 21-hydroxylase deficiency. PMID- 12560362 TI - Rapid HPLC-electrospray tandem mass spectrometric assay for urinary testosterone and dihydrotestosterone glucuronides from patients with benign prostate hyperplasia. PMID- 12560363 TI - Simple PCR-based method for synthesis of molecular calibrators and controls. PMID- 12560364 TI - Protein glutathionylation in erythrocytes. PMID- 12560365 TI - Adaptation of a mitochondrial complex III assay for automation: examination of reproducibility and precision. PMID- 12560366 TI - Impact of assay epitope specificity in gastrinoma diagnosis. PMID- 12560367 TI - Use of microsphere immunoassay for simplified multianalyte screening of thyrotropin and thyroxine in dried blood spots from newborns. PMID- 12560368 TI - Comparison of two third-generation parathyroid hormone assays. PMID- 12560370 TI - Improved method for measurement of retinol and didehydroretinol in the modified relative dose response test to detect vitamin A deficiency. PMID- 12560371 TI - Effects of blood collection methods on gelatin zymography of matrix metalloproteinases. PMID- 12560372 TI - Effect of sulfamethoxazole on clinical capillary zone electrophoresis of serum proteins. PMID- 12560383 TI - acp Best Practice No 169. Evidence based pathology: squamous carcinoma of the hypopharynx. AB - This best practice article reviews the published evidence on the pathology and patterns of spread of carcinomas of the hypopharynx, and the relevance of pathological features to prognosis. Medline (1966-2001) was searched using a combination of head and neck neoplasms and prognosis, focusing on hypopharynx and pathology. Other relevant publications were identified from the bibliographies of these papers, and from those obtained opportunistically. There is relatively little pathological literature devoted specifically to squamous carcinomas of the hypopharynx and most information comes from large series of patients with head and neck cancers at a range at sites. Lack of consistency in reporting and shifts in terminology make comparisons between series difficult. The most important features determining prognosis are size and extent of local spread of the primary carcinoma and extent of involvement of regional lymph nodes. There is evidence to support the use of the minimum dataset criteria for head and neck carcinomas at this site. Within the hypopharynx, subsite related differences in aetiology and biology may become important. PMID- 12560384 TI - Micropurification techniques in the analysis of amyloid proteins. AB - This review describes the different microtechniques developed for the extraction and purification of amyloid proteins from small specimens of fresh and formalin fixed tissues. These procedures differ with respect to solvent type, extraction conditions, and protein purification strategy. The advantages and disadvantages of the different microtechniques are discussed by taking into consideration tissue type (fresh of fixed) and size, amyloid type, and its content in the tissue. The review demonstrates the applicability of these techniques for the immunochemical and chemical characterisation of amyloid in different clinical forms of amyloidosis and in experimental small animal models. The clinical value of the applied microtechniques and their importance in the study of the pathogenesis of amyloid related diseases are outlined. PMID- 12560385 TI - Classification and grading of the non-invasive urothelial neoplasms: recent advances and controversies. AB - The classification and grading of the non-invasive, intraepithelial neoplasms of the urothelium are based on the morphological pattern of growth-that is, papillary or flat (and endophytic)-and on their degree of architectural and cytological abnormalities. Recent advances in the morphological, molecular, and quantitative evaluation of these lesions have contributed to the refinement of the current classification and grading schemes. However, some controversies on the precise criteria and terminology, especially when the papillary lesions are concerned, are still present. PMID- 12560386 TI - Ewing's sarcoma: diagnostic, prognostic, and therapeutic implications of molecular abnormalities. AB - The identification of the non-random chromosome rearrangements between the EWS gene on chromosome 22q12 and members of the ETS gene family in Ewing's sarcoma, peripheral primitive neuroectodermal tumour, Askin tumour, and neuroepithelioma has been a key advance in understanding their common histogenesis and defining the Ewing's sarcoma family of tumours (ESFT). In addition to improvements in diagnosis and potentially the stratification of patients for risk, biological investigations of these gene fusions may define targets for much needed therapeutic strategies to eliminate minimal residual disease or metastatic disease. Insight into their relation with other oncogenic events in ESFT will advance risk group analysis and ultimately may improve clinical management and survival for patients with this disease. PMID- 12560387 TI - Chymase expressing bone marrow mast cells in mastocytosis and myelodysplastic syndromes: an immunohistochemical and morphometric study. AB - BACKGROUND: Two cell specific neutral proteases, tryptase and chymase, are produced by human mast cells (MC). Tryptase is constitutively expressed by all MC, whereas chymase is found only in an MC subset. Very little is known about chymase expression in MC proliferative disorders (mastocytosis). AIMS AND METHODS: Routinely processed, formalin fixed, and paraffin wax embedded bone marrow trephine biopsy specimens obtained from patients with various subtypes of mastocytosis (n = 47) and myelodysplastic syndromes (MDS; n = 28) were immunostained with antibodies against chymase and tryptase. Normal/reactive bone marrow specimens with intact haemopoiesis (n = 31) served as controls. The numbers of chymase expressing (C+) and of tryptase expressing (T+) MC were assessed morphometrically using a computer assisted video camera system. RESULTS: In normal/reactive bone marrow, the numbers of C+ MC (median, 8/mm(2); maximum, 159/mm(2)) were in the same range as those of T+ MC (median, 4/mm(2); maximum, 167/mm(2)). Because normal MC express both chymase and tryptase, these findings indicate that the common phenotype of bone marrow MC in normal/reactive states is MC(TC) (MC expressing both tryptase and chymase). In contrast, in MDS and mastocytosis, the bone marrow exhibited far more T+ MC than C+ MC in almost all cases. CONCLUSIONS: According to these findings, the predominant MC type in the bone marrow in neoplastic states such as MDS and mastocytosis is MC(T) (MC expressing only tryptase). Although the pathophysiological basis of this apparent lack of chymase expression in most neoplastic MC in mastocytosis and MC involved in MDS remains unknown, this study has produced further evidence of the superior value of antitryptase antibodies in the diagnosis of mastocytosis. PMID- 12560388 TI - Expression and prognostic relevance of vascular endothelial growth factor (VEGF) and its receptor (FLT-1) in nephroblastoma. AB - AIMS: To investigate the prognostic relevance of vascular endothelial growth factor (VEGF) and its receptor Flt-1 in nephroblastoma and whether tumour microvessel density (MVD) immunoreactivity, determined by the CD31 antigen, is related to the expression of VEGF and Flt-1. METHODS: The expression of VEGF and Flt-1 and MVD were investigated by means of immunohistochemical analysis in 62 Wilms's tumours. Patients were treated preoperatively with chemotherapy and had a mean follow up of 5.7 years. RESULTS: In general, VEGF and Flt-1 were expressed in normal kidney parenchyma and to a variable extent in the three main components of Wilms's tumour, namely: the blastemal, epithelial, and stromal cells. In tumour tissue, 52% and 47% of blastemal cells were positive for VEGF and Flt-1, respectively. A non-significant correlation was found between the expression of VEGF and Flt-1 in blastemal and epithelial cells and the clinicopathological stage. MVD was significantly higher in VEGF and Flt-1 positive tumours than in VEGF and Flt-1 negative tumours. Univariate analysis showed that the expression of VEGF and Flt-1 in blastemal cells was indicative of clinical progression and tumour specific survival. In addition, MVD expression was indicative of clinical progression. Epithelial staining was of no prognostic value. In a multivariate analysis, VEGF protein expression by blastemal cells was an independent prognostic marker for clinical progression. CONCLUSIONS: These results indicate that VEGF and Flt-1 protein expression are closely related to MVD and seem to be an important predictor for poor prognosis in treated patients with Wilms's tumour. Therefore, the expression of these molecules in primary Wilms's tumour may be useful in identifying those patients at high risk of tumour recurrence and in guiding antiangiogenic treatment. PMID- 12560389 TI - European Concerted Action on Anticoagulation (ECAA). An assessment of lyophilised plasmas for ISI calibration of CoaguChek and TAS whole blood prothrombin time monitors. AB - AIMS: The recommended method for the international sensitivity index (ISI) calibration of whole blood point of care testing (POCT) prothrombin time (PT) systems was originally described by Tripodi et al in 1993 but is too complex and demanding. The present European Concerted Action on Anticoagulation (ECAA) study aimed to assess the reliability of simpler ISI calibration using lyophilised plasma samples. METHODS: ISI calibrations using three different types of ECAA lyophilised plasma samples (artificially depleted, individual, and pooled coumarin) were compared with whole blood calibrations on CoaguChek Mini and TAS PT-NC POCT monitors at 10 centres. RESULTS: With CoaguChek Mini systems, lyophilised coumarin plasma samples (both single donation and pooled) gave ISI and international normalised ratio (INR) values comparable to whole blood. With artificially depleted plasma, ISI and INR values were too high. With TAS PT-NC systems, all three types of lyophilised plasma samples gave inaccurate ISI and unreliable INR results, similar to previous ECAA findings with fresh plasma calibrations. CONCLUSIONS: With CoaguChek Mini systems, ISI calibration can be simplified by the use of ECAA lyophilised plasma samples from coumarin treated patients. Further study is needed to devise a simpler calibration method for the TAS PT-NC system. PMID- 12560390 TI - Genetic characterisation of adenovirus type 8 isolated in Hiroshima city over a 15 year period. AB - AIMS: To investigate the genetic differences among the strains of adenovirus type 8 (Ad8) circulating in Hiroshima city, Japan, and to study their circulation pattern. METHODS: One hundred and twenty nine strains of adenovirus type 8 (Ad8) were isolated in Hiroshima City over a 15 year period (1983-97) from patients with keratoconjunctivitis, and analysed with six restriction enzymes-BamHI, HindIII, PstI, SacI, SalI, and SmaI-to investigate possible relations among the isolates and their genetic variability. Seven hypervariable regions of the hexon gene that carry the type specific epitope were also sequenced to investigate the variation among the genome types. RESULTS: Restriction endonuclease analyses yielded three known genome types (Ad8A, 13 samples; Ad8B, seven samples; and Ad8E, 35 samples) and a novel genome type (Ad8I, 74 samples). Ad8A, Ad8B, and Ad8E were closely related, with 96% homology, whereas Ad8I had only 71% homology. Ad8A, Ad8B, and Ad8E shared 91.8% and 96.4% homology with regard to their amino acid and nucleotide sequences, respectively, with the isolate 1127 (accession no X74663). However, when compared with Ad8A, Ad8B, Ad8E, and isolate 1127, Ad8I shared only 62.7% and 69.9% homology with regard to amino acid and nucleotide sequences, respectively. Ad8A, Ad8B, and Ad8E had a unique 31 amino acid deletion in the hypervariable region 1 of the hexon gene, whereas Ad8I had a 33 residue deletion. The Ad8E strain that circulated from 1984 to 1995 was stable among the study population. Ad8I was isolated from an outbreak of epidemic keratoconjunctivitis in 1995 and was also isolated from sporadic cases until 1997. CONCLUSIONS: These results confirmed that genetic variability occurs in Ad8 in the microenvironment and revealed the emergence of a new genome type (Ad8I). PMID- 12560391 TI - The effects of storage conditions on viability of Clostridium difficile vegetative cells and spores and toxin activity in human faeces. AB - AIMS: Clostridium difficile is a common nosocomial pathogen and as such diagnostic and research methods may necessitate storage of faecal specimens for long periods, followed by subsequent re-examination. This study investigated the effects of storage conditions upon the viability of this organism and its toxin. METHODS: Three genotypically distinct strains of C difficile (two clinical isolates including the UK epidemic strain, and an environmental isolate) were grown anaerobically at 37 degrees C for 72 hours in a pool of five faecal emulsions. Aliquots of each emulsion were stored at either -20 degrees C (frozen) or 4 degrees C (refrigerated). Emulsions were assayed for viable cells, spores, and cytotoxin titre before storage and at days 1, 3, 5, 7, 14, 28, and 56. An aliquot of each emulsion was also removed, assayed, and replaced in storage at each time point to investigate the effects of multiple freezing/refrigeration/thawing. RESULTS: Neither storage temperature nor multiple cycles of refrigeration/freezing and thawing adversely affected the viability of C difficile vegetative cells or spores. Single and multiple exposures of samples to 4 degrees C had little effect upon the C difficile toxin titre. Toxin titres of multiply frozen and thawed faeces became significantly lower than for refrigerated faeces (p < 0.01) by day 5 of the experiment in two of the three strains, and in all strains by day 28. Toxin titres of singly frozen faeces became significantly lower than for refrigerated faeces (p < 0.01) by day 56 of the experiment in two of the three strains. CONCLUSION: Storage temperature and multiple cycles of freezing (refrigeration)/thawing had minimal effects upon the viability of C difficile or its spores. Storage at 4 degrees C has no discernible effect on C difficile cytotoxin. However, storage at -20 degrees C has a detrimental effect upon C difficile cytotoxin, and multiple cycles of freezing and thawing may further adversely effect toxin titres. PMID- 12560392 TI - A comparison of flow cytometry, bone marrow biopsy, and bone marrow aspirates in the detection of lymphoid infiltration in B cell disorders. AB - AIMS: To evaluate the diagnostic value of bone marrow aspirates, trephine biopsies (BMB), and flow cytometry (FC) in the assessment of bone marrow infiltration in chronic lymphoid disorders. METHODS: Investigations were carried out in 110 diagnostic and follow up specimens from B cell disorders, namely: chronic lymphocytic leukaemia (CLL; 65), non-Hodgkin's lymphoma (NHL; 39), and hairy cell leukaemia (HCL; 6). A selected panel of monoclonal antibodies was used both for FC and immunohistochemistry. RESULTS: In CLL there was agreement between the three investigations in 71% of samples and in 88% when only FC and BMB were compared. In nine of 65 samples, FC and BMB were positive, although the aspirate was reported as negative. Four BMB negative samples had minimal residual disease (MRD) detected by FC, whereas two samples were positive both on BMB and aspirate but showed no evidence of disease on FC. In NHL, there was agreement between the three investigations in 22 of 39 cases, and in 27 of 39 cases there was agreement between FC and BMB. In eight of 39 NHL cases, FC was negative but the BMB was either positive (five) or uncertain (three), whereas in three of 39, FC was positive but BMB was either negative (one) or uncertain (two). In three of five uncertain BMB, no clonal population was detected by the polymerase chain reaction, whereas in the remaining two cases the nodular aggregates disappeared on further sectioning. CONCLUSIONS: Both BMB and FC are better than bone marrow aspirates for the detection of infiltration in B cell disorders. FC might be slightly more sensitive than BMB to detect MRD in CLL, whereas BMB may be slightly better than FC in NHL. PMID- 12560393 TI - Will the spectrum of lesions prompting a "B3" breast core biopsy increase the benign biopsy rate? AB - AIM: To audit the benign surgical biopsies in women screened, assessed, and referred by the Leeds/Wakefield Breast Screening Unit for the year 1999-2000 with a view to determining any association with a preoperative B3 core biopsy categorisation. METHODS: The results of all preoperative diagnostic procedures in all patients who underwent surgical excision for a lesion proving benign in the year 1999-2000 were reviewed. Cases were categorised according to whether the preoperative fine needle aspirate cytology (FNAC) or core biopsy had been equivocal or of uncertain malignant potential (C3/B3), inadequate or unrepresentative (C1/B1), or benign (C2/B2). In those cases with a C3/B3 FNAC or core biopsy result, reasons for the uncertainty were determined by examination of the report and, where necessary, slides. In cases with C1/B1 or C2/B2 investigations and in those without a preoperative procedure, the reasons for surgical referral were determined from the screening records. Case records of all patients with a B3 core biopsy categorisation who subsequently proved to have malignancy were also reviewed. RESULTS: Thirty six women had benign surgical biopsies in the 1999-2000 screening year. In 13 of the 36 patients, referral for diagnostic biopsy rested on radiological and/or pathological suspicion of radial scar. The core biopsy category was B3 in all but one, which was in the B1 category. In a further 10 patients, referral was based primarily on a pathological B3 categorisation. The reasons for this were as follows: papillary lesion (two), fibroepithelial lesion (two), atypical intraductal epithelial proliferation (two), stromal mucin (two), atypical lobular hyperplasia (one), and an unusual vascular lesion (one). Two cases with a C3 on FNAC also derived from papillary lesions. In the remaining nine patients, the radiological features were sufficiently suspicious to prompt referral in the presence of either inadequate/unrepresentative (C1/B1) or benign (B2) preoperative pathological findings. Two women had no preoperative needle biopsy. CONCLUSIONS: In 22 of 36 benign biopsies, the initial core biopsy categorisation was B3. According to the current system of core biopsy categorisation, a diversity of lesions must be designated as of "uncertain malignant potential" (B3) because the technique provides insufficient tissue for full histological assessment. The use of this category may increase the number of benign biopsies if all such cases are referred for surgery. An increase in the benign biopsy rate may be averted if larger amounts of tissue can be obtained using newer vacuum assisted techniques such as the Mammotome. PMID- 12560394 TI - P-cadherin as a marker in the differential diagnosis of breast lesions. AB - AIMS: To assess the value of the calcium dependent cell adhesion molecule P cadherin as a myoepithelial marker in the differential diagnosis of benign and malignant breast lesions. METHODS: Immunohistochemical analysis of normal breast, sclerotic breast lesions, tubular carcinomas, and ductal carcinoma in situ using a P-cadherin specific antibody and comparison with smooth muscle actin. RESULTS: All myoepithelial cells in normal breast ducts, ductules, and lobules and sclerotic lesions showed strong staining for P-cadherin. There was no staining of tubular carcinomas; myoepithelial cells were demonstrated around in situ carcinomas. Weaker reactivity was seen in a proportion of cells in some hyperplasias and in situ carcinomas. This weak reactivity in these tissues was not seen for smooth muscle actin but in radial scars, tubular carcinomas, and ductal carcinoma in situ staining of stromal cells caused difficulties in the identification of myoepithelial cells. CONCLUSION: P-cadherin is a useful marker in the differential diagnosis of breast lesions. PMID- 12560396 TI - Differential expression of galectin 3 in solid cell nests and C cells of human thyroid. AB - AIMS: To provide new insights into characterising solid cell nests and gain information that might help distinguish between solid cell nests and C cells. METHODS: Thyroid tissue specimens from patients who had undergone prophylactic thyroidectomy for familial medullary thyroid cancer were immunostained for calcitonin, carcinoembryonic antigen, and galectin 3. RESULTS: Solid cell nests displayed a strong and diffuse staining for carcinoembryonic antigen and galectin 3, but not for calcitonin. C cells located at the periphery of solid cell nests and in neighbouring follicles expressed both calcitonin and carcinoembryonic antigen but not galectin 3. These three markers were positive in medullary thyroid carcinoma. CONCLUSION: Galectin 3 immunoreactivity permits a better characterisation and differentiation between solid cell nests and C cells, avoiding the misidentification of two biologically and clinically different thyroid structures. PMID- 12560397 TI - A simple method for the construction of small format tissue arrays. AB - Tissue arrays can evaluate molecular targets in high numbers of samples in parallel. Array construction presents technical difficulties and tissue arrayers are expensive, particularly for small and medium sized laboratories. This report describes a method for the construction of 36 sample arrays using widely available materials. A blunted 16 gauge needle for bone marrow aspiration was used to extract paraffin wax cylinders and manually define a 6 x 6 matrix on a blank paraffin wax block. Tissue cores from 36 paraffin wax embedded premalignant lesions and invasive cervical carcinomas were injected into the matrix using a 14 gauge needle. This tissue array was sectioned using a standard microtome and used for the immunodetection of CD44 variant 9 and interleukin 18 with satisfactory results. This method can be applied in any laboratory, without the need of specialised equipment, offering a good alternative for the wider application of tissue arrays. PMID- 12560398 TI - Mitochondrial disruption and apoptosis in lymphocytes of an HIV infected patient affected by lactic acidosis after treatment with highly active antiretroviral therapy. AB - AIMS: Highly active antiretroviral therapy (HAART) can induce an increase in lactic acid concentrations that seems to be caused by mitochondrial dysfunction induced by the interaction of nucleoside reverse transcriptase inhibitors (NRTIs) with DNA polymerase gamma in the mitochondria. Mitochondrial alterations have been described in liver and muscle cells of NRTI treated human immunodeficiency virus (HIV) infected patients. Because lymphocytes are the main target for HIV and because mitochondria are involved in apoptosis, we studied mitochondrial morphology and apoptosis in the lymphocytes of an HIV infected patient with severe lactic acidosis after treatment with stavudine, didanosine, and indinavir. METHODS: The patient was a 39 year old woman. After two years of treatment she developed rapid weight loss with severe fat wasting, peripheral neuropathy, and hyperlacticaemia, which persisted after treatment withdrawal. The numbers and the morphology of the mitochondria were evaluated by electronic microscopy; the percentage of apoptotic cells was calculated by flow cytometry after staining with annexine V and by fluorescent microscopy after staining with ethidium bromide and acridine orange. RESULTS: The numbers of mitochondria in the lymphocytes were greatly decreased when compared with the lymphocytes of healthy individuals. The most important mitochondrial morphological alterations were swelling and the disruption of cristae and internal mitochondrial structure. These alterations were more evident during the period in which lactic acid values were very high. Moreover, a high percentage of apoptotic lymphocytes was seen. Morphological examination conducted one week after the normalisation of lacticaemia showed a pronounced increase in the number of mitochondria. The morphological alterations were no longer evident, although the size of each mitochondrion was smaller than normal. Moreover, the percentage of apoptotic cells was lower than 5%. CONCLUSIONS: This report describes important morphological alterations in lymphocyte mitochondria in an HIV infected patient during a severe phase of HAART induced hyperlacticaemia. These alterations persisted for several weeks after treatment withdrawal and were associated with an increase in lymphocyte apoptosis. Considering the important role of mitochondria in the apoptotic pathway, the increase in lymphocyte apoptosis may be a consequence of proapoptotic factors released from altered mitochondria. PMID- 12560399 TI - Urachal adenocarcinoma in situ with pseudomyxoma peritonei: a case report. AB - A 54 year old man presented with a six month history of abdominal pain. A computerised tomography scan showed a well defined intra-abdominal unilocular mass with a calcified wall just superior to the bladder. At laparotomy, pseudomyxoma peritonei was discovered, together with a midline abdominal mass adherent to the anterior abdominal wall originating from the fundus of the bladder. The specimen consisted of a cystic mass measuring 14 x 9.5 x 7 cm overall, which contained mucoid material. Histological examination revealed that the cyst was lined by mucinous epithelium, which in areas varied from having bland morphology to showing pronounced nuclear and architectural atypia. There was abundant extracellular mucin. The specimen was extensively sampled but there was no evidence of invasion. This tumour has many unusual features, namely: the absence of destructive invasion, association with pseudomyxoma peritonei, areas of dysplasia and cystadenoma, and stromal osseous metaplasia within the wall. PMID- 12560400 TI - Myelofibrosis presenting as spinal cord compression. AB - This report describes a case of myelofibrosis presenting as spinal cord compression on account of extramedullary haemopoietic tissue encroaching upon the spinal cord from a large pelvic mass. PMID- 12560401 TI - Molten gold was poured down his throat until his bowels burst. PMID- 12560402 TI - The number of intraepithelial T cells decreases from ascending colon to rectum. PMID- 12560403 TI - Food for thought. PMID- 12560404 TI - How do we define Hodgkin's disease? The authors' reply. PMID- 12560405 TI - Morphological and functional characteristics of skeletal muscle fibers from hormone-replaced and nonreplaced postmenopausal women. AB - We tested the hypothesis that cross-bridge mechanisms of contraction differed in early postmenopausal women who did or did not receive hormone replacement therapy (HRT). Vastus lateralis biopsies were obtained from 17 postmenopausal women (49 57 years old), 8 of whom were on HRT for the previous 24 +/- 5 months and 9 of whom were never on HRT. Electrophoresis and enzyme histochemistry revealed that fiber myosin heavy chain (MHC) isoform distribution, the cross-sectional area (CSA) of slow and fast fibers, and the relative CSA occupied by each, were similar for HRT and non-HRT groups. Single permeabilized fibers containing type IIa MHC had greater Ca(2+)-activated peak specific force, unloaded shortening velocity, and peak power than fibers containing type I MHC, but in all cases the values for HRT and non-HRT groups were similar. In this cross-sectional study, we found no evidence that Ca(2+)-activated fiber function, MHC isoform distribution, or relative CSA occupied by slow and fast fibers differed between HRT and non-HRT groups. PMID- 12560406 TI - Creatine supplementation enhances isometric strength and body composition improvements following strength exercise training in older adults. AB - We sought to determine whether creatine monohydrate (CrM) supplementation would enhance the increases in strength and fat-free mass that develop during resistance exercise training in older adults. Twenty-eight healthy men and women over the age of 65 years participated in a whole-body resistance exercise program 3 days per week for 14 weeks. The study participants were randomly allocated, in a double-blind fashion, to receive either CrM (5 g/d + 2 g of dextrose; n = 14) or placebo (7 g of dextrose; n = 14). The primary outcome measurements included the following: total body mass, fat-free mass, one-repetition maximum strength for each body part, isometric knee extension, handgrip, and dorsiflexion strength, chair stand performance, 30-m walk test, 14-stair climb performance, muscle fiber type and area, and intramuscular total creatine. Fourteen weeks of resistance exercise training resulted in significant increases in all measurements of strength and functional tasks and muscle fiber area for both groups (p <.05). CrM supplementation resulted in significantly greater increases in fat-free mass and total body mass, as compared with placebo (p <.05). The CrM group also showed a greater increase in isometric knee extension strength in men and women, as compared with placebo (p <.05), and also greater gains in isometric dorsiflexion strength (p <.05), but in men only. There was a significant increase in intramuscular total creatine in the CrM group (p <.05). Finally, there were no significant side effects of treatment or exercise training. This study confirms that supervised heavy resistance exercise training can safely increase muscle strength and functional capacity in older adults. The addition of CrM supplementation to the exercise stimulus enhanced the increase in total and fat free mass, and gains in several indices of isometric muscle strength. PMID- 12560407 TI - Lower skeletal muscle nutritive blood flow in older women is related to eNOS protein content. AB - The relationship between muscle endothelial nitric oxide synthase (eNOS) content and nutritive flow was investigated in nonobese sedentary young (27.7 +/- 2.6 years) and older (56.6 +/- 2.1 years) women matched for body composition and (2)peak. A muscle biopsy was taken and nutritive blood flow was determined under resting conditions in the vastus lateralis of the quadriceps femoris muscle group. Muscle eNOS protein content correlated with muscle nutritive blood flow (r =.66, p <.05) and body mass index (r =.74, p <.05), but it did not correlate with VO(2)peak. Muscle eNOS content was 35% lower in young than in older women (266 +/ 36 vs 407 +/- 53 pg/mg total protein; p <.05). The mean ethanol outflow-to inflow ratio was higher (indicating lower nutritive flow) in older and young women (.666 +/-.042 and.546 +/-.043, respectively: p <.05). Resting skeletal nutritive blood flow and muscle eNOS content was lower in older than in young women. A low muscle eNOS protein content may be linked to a low muscle nutritive blood flow in healthy women. PMID- 12560408 TI - Hot topics in geriatrics. PMID- 12560409 TI - Cognitive screening predicts magnitude of functional recovery from admission to 3 months after discharge in hospitalized elders. AB - BACKGROUND: Many older adults lose functional ability during the course of acute illness and fail to recover function. We sought to determine whether performance on a cognitive screen at the time of hospital admission predicted the magnitude of functional recovery after hospitalization. METHODS: We studied 2557 patients from two teaching hospitals to examine the association between level of impaired performance on a cognitive status screen and maintenance and recovery of functioning from admission through 90 days after discharge. On admission, 14% had mildly impaired cognitive performance with three or four errors on the Short Portable Mental Status Questionnaire; 28% had moderate to severely impaired cognitive performance with five or more errors on the cognitive status screen or inability to complete the screen and a diagnosis of dementia. RESULTS: Performance on a brief cognitive screen on admission was strongly related to subsequent change in function. Among patients who needed help performing one or more activities of daily living at the time of admission, 23% of patients with moderate to severely impaired cognitive performance, 49% of patients with mildly impaired cognitive performance, and 67% of patients with little to no impairment in cognitive performance recovered ability to independently execute an additional activity of daily living by discharge (p <.001). Similar relationships were seen for change in instrumental activities of daily living and mobility. In multivariate repeated measures analyses of basic and instrumental activities of daily living and mobility on admission, discharge, and 30 and 90 days after discharge, patients with mildly impaired cognitive performance on admission showed less improvement than patients who did not have impaired cognitive performance, but more than those with moderate to severely impaired cognitive performance. The pattern of results did not change when patients with any signs of delirium were excluded. Patients with impaired cognitive performance were more likely to be admitted to a nursing home for the first time by 90 days after discharge. The odds ratios were 2.8 (95% confidence interval = 1.8-4.5) for patients with mildly impaired cognitive performance and 6.7 (95% confidence interval = 4.5-9.8) for patients with moderate to severely impaired cognitive performance. CONCLUSION: Cognitive screening at hospital admission can be used to stratify patients according to the magnitude of expected functional recovery after an acute illness that required hospitalization. PMID- 12560410 TI - Step training improves the speed of voluntary step initiation in aging. AB - BACKGROUND: Falls related to balance dysfunction are among the major problems of older individuals. The timing characteristics of protective voluntary stepping are critically related to effective balance recovery and are often delayed and slowed with age. This study investigated the influence of step training on the timing characteristics of voluntary step initiation in younger and older adults. METHODS: Voluntary reaction time stepping was evaluated before and after training in 12 younger adults and 8 healthy community-dwelling older adults who performed a 3-week regimen of either twice weekly induced step training (destabilizing large waist pulls) or voluntary step practice to a somatosensory reaction stimulus cue (nondestabilizing small waist pulls). RESULTS: Overall, the first step initiation times were slower for the older than for the younger subjects for both the somatosensory reaction stimulus cue task and an auditory transfer cue task. Step completion time was completed earlier for the young posttraining subjects, and older subjects generally had a longer step length. Training resulted in significant reductions in step initiation timing for the old (17%) and young (15%) subjects. Across age groups, the induced training group showed greater reductions in step initiation time than the voluntary practice group for the auditory transfer cue task. CONCLUSIONS: A 3-week period of either voluntary or waist-pull-induced step training reduced step initiation time in older and younger adults. Moreover, compared with voluntary step practice, induced step training resulted in a significantly greater improvement in reaction time stepping for an auditory transfer cue task. At least in the short term, such step training has the potential to help older adults perform more like younger adults in their step initiation timing. PMID- 12560411 TI - Pathogenic colonization of oral flora in frail elderly patients fed by nasogastric tube or percutaneous enterogastric tube. AB - BACKGROUND: Aspiration of infected oropharyngeal content is the main cause of aspiration pneumonia. This complication, mainly related to gram-negative bacteria, threatens percutaneous enterogastric tube as well as nasogastric tube (NGT) fed patients. The objective of this study was to examine the oral microbiota of tuboenterally fed patients and compare it with that of orally fed counterparts. METHODS: Patients were recruited for this study from six nursing and skilled nursing facilities with an overall number of 845 beds. Enrolled were 215 patients: Group 1 consisted of 78 patients on NGT feeding, Group 2 consisted of 57 patients on percutaneous enterogastric tube feeding, and Group 3 consisted of 80 patients fed orally who were from the same facilities. Cultures were performed by sampling the oropharynx of each subject in order to identify gram negative bacteria and Staphylococcus aureus. RESULTS: A high prevalence of potentially pathogenic isolations was found in tuboenterally fed patients: 81% in Group 1 and 51% in Group 2, as compared with only 17.5% in Group 3 (p <.0001). Pseudomonas aeruginosa was cultured from 31% of the subjects in Group 1 and 10% of Group 2, but in none of Group 3 (p <.001). Klebsiella and Proteus were isolated mainly from the NGT fed patients (p <.003). No correlation was found between the time duration on tube feeding or the presence of residual dentition and pathogenic microbiota. CONCLUSION: This study shows that tuboenteral feeding in elderly patients is associated with pathogenic colonization of the oropharynx. These findings are related to the risk of aspiration pneumonia and are compelling for the reevaluation of current oral cleansing procedures. PMID- 12560412 TI - Elderly hospitalized patients with diastolic heart failure: lack of gender and ethnic differences in 18-month mortality rates. AB - BACKGROUND: Racial and gender differences in mortality rates have been reported for patients with systolic heart failure. Relatively little is known regarding diastolic heart failure prognosis. METHODS: Our sample consisted of 1058 patients 65 years of age or older who were admitted to 30 hospitals in Northeastern Ohio with a principal diagnosis of heart failure and a left ventricular ejection fraction of >/=50% by echocardiogram. RESULTS: Of the 1058 patients with diastolic heart failure (13% African American and 87% white), African Americans and whites were comparable with respect to history of angina, stroke, being on dialysis, and alcohol use; the proportion of male patients was also comparable. The African American to white adjusted odds ratio for 18-month mortality (all cause) was 1.03 (0.66-1.59). For men versus women (30% vs 70%), the above mentioned comorbidities were comparable, except women were more likely to have a do not resuscitate status (16% vs 7.3%; p =.000) and to be older (79.5 +/- 8 vs 77 +/- 7; p =.000). Men were more likely to have a history of tobacco use (30% vs 14%; p =.000) and alcohol use (36% vs 15%; p =.000), and a higher serum creatinine level (1.7 +/- 1.2 vs 1.4 +/- 1.1; p =.001). The men to women adjusted odds ratio for 18-month mortality (all cause) was 1.06 (0.76-1.46). CONCLUSION: In this cohort of elderly patients admitted with diastolic heart failure, there were no ethnic or gender differences in 18-month mortality rates. PMID- 12560413 TI - Predictors of short-term functional decline in survivors of nursing home-acquired lower respiratory tract infection. AB - BACKGROUND: Scant information exists about the risk of functional decline following treatment of acute illness in the nursing home (NH) setting. The aim of this study was to determine the incidence of short-term (30-day) functional decline among survivors of NH-acquired lower respiratory tract infection (LRI) and the factors that predict such decline, including the role of initial hospitalization. METHODS: We used a prospective cohort design to study 781 episodes of LRI in 1044 NH residents in 36 NHs in central Missouri and the St. Louis metropolitan area. Functional decline was defined as a 3-point increase on the Minimum Data Set (MDS) activities of daily living (ADL) long form scale. RESULTS: Of 781 LRI cases who survived to 30 days, the incidence of ADL decline was 28.8%. In a logistic regression model that used generalized estimating equations to adjust for clustering, variables associated with ADL decline included the following: chronic feeding tube use (AOR = 4.54, 95% confidence interval, or CI, 1.61, 12.80), decubitus ulcer (adjusted odds ratio [AOR] = 2.29, 95% CI 1.35, 3.90), shortness of breath (AOR = 2.18, 95% CI 1.44, 3.30), short term memory problems (AOR = 2.07, 95% CI 1.33, 3.23), decline in self-performance of toilet use in the 24 hours prior to evaluation (AOR = 1.65, 95% CI 1.29, 2.12), age (AOR = 1.02, 95% CI 1.00, 1.05), and baseline ADL score. Addition of treatment variables to the model showed that initial hospitalization was also associated with ADL decline (AOR = 1.90, 95% CI 1.20, 3.00). Residents with ADL decline at 30 days were less likely to recover to their baseline ADL status at 90 days. CONCLUSIONS: Many NH residents who survive to 30 days following LRI develop new functional limitations, and such individuals are at risk for ADL decline at 90 days. A limited number of clinical variables may predict short-term functional decline. Initial hospitalization for acute treatment of LRI may increase the risk of subsequent ADL decline among individuals who survive to 30 days. PMID- 12560414 TI - Increased prevalence of peripheral arterial disease in older men and women with subclinical hypothyroidism. AB - BACKGROUND: We report the prevalence of symptomatic peripheral arterial disease (PAD) associated with subclinical hypothyroidism in older persons. METHODS: We investigated the association of subclinical hypothyroidism with symptomatic PAD in 18 persons (16 women and 2 men) with subclinical hypothyroidism and in 231 euthyroid persons (151 women and 96 men), mean age 79 +/- 9 years, in an academic nursing home. RESULTS: Symptomatic PAD was present in 14 of 18 persons (78%) with subclinical hypothyroidism and in 40 of 231 euthyroid persons (17%; p <.0001, comparing subclinical hypothyroidism with euthyroidism). Symptomatic PAD was present in 13 of 16 women (81%) and in 1 of 2 men (50%) with subclinical hypothyroidism (p was not significant). CONCLUSIONS: Subclinical hypothyroidism was associated with a high prevalence of symptomatic PAD in elderly men and women. PMID- 12560415 TI - The burden and patterns of disability in activities of daily living among community-living older persons. AB - BACKGROUND: The onset of disability in activities of daily living (ADLs) is considered a sentinel event in the life of an older person, but recent evidence suggests that newly disabled elders have high rates of recovery. We performed a longitudinal study to determine the burden and patterns of ADL disability among previously nondisabled, community-living older persons. METHODS: We studied 754 community-living persons, aged 70 years or older, who were categorized into three groups according to their risk for disability (low, intermediate, high). Participants were interviewed each month for 2 years to determine the presence and severity of disability in four key ADLs: bathing, dressing, walking, or transferring. RESULTS: Among the 690 nondecedents, the rates of any disability were 17.7%, 48.7%, and 65.2%, respectively, for the low-, intermediate-, and high risk groups. Whereas only 6.9% of nondecedents in the low-risk group had more than 1 month of disability, 38.2% and 50.6% of nondecedents in the intermediate- and high-risk groups (i.e., physically frail), respectively, had multiple months and/or episodes of disability. The patterns of disability were quite diverse, with no single pattern representing the disability experience of more than half the decedents or nondecedents in any of the risk groups. CONCLUSIONS: Disability among community-living older persons, particularly those who are physically frail, is a highly dynamic process with considerable diversity. Our results provide strong evidence to support an emerging paradigm of disability as a reversible, and often recurrent, event. PMID- 12560416 TI - Relationship between pain and opioid analgesics on the development of delirium following hip fracture. AB - BACKGROUND: Delirium and pain are common following hip fracture. Untreated pain has been shown to increase the risk of delirium in older adults undergoing elective surgery. This study was performed to examine the relationship among pain, analgesics, and other factors on delirium in hip fracture patients. METHODS: We conducted a prospective cohort study at four New York hospitals that enrolled 541 patients with hip fracture and without delirium. Delirium was identified prospectively by patient interview supplemented by medical record review. Multiple logistic regression was used to identify risk factors. RESULTS: Eighty-seven of 541 patients (16%) became delirious. Among all subjects, risk factors for delirium were cognitive impairment (relative risk, or RR, 3.6; 95% confidence interval, or CI, 1.8-7.2), abnormal blood pressure (RR 2.3, 95% CI 1.2 4.7), and heart failure (RR 2.9, 95% CI 1.6-5.3). Patients who received less than 10 mg of parenteral morphine sulfate equivalents per day were more likely to develop delirium than patients who received more analgesia (RR 5.4, 95% CI 2.4 12.3). Patients who received meperidine were at increased risk of developing delirium as compared with patients who received other opioid analgesics (RR 2.4, 95% CI 1.3-4.5). In cognitively intact patients, severe pain significantly increased the risk of delirium (RR 9.0, 95% CI 1.8-45.2). CONCLUSIONS: Using admission data, clinicians can identify patients at high risk for delirium following hip fracture. Avoiding opioids or using very low doses of opioids increased the risk of delirium. Cognitively intact patients with undertreated pain were nine times more likely to develop delirium than patients whose pain was adequately treated. Undertreated pain and inadequate analgesia appear to be risk factors for delirium in frail older adults. PMID- 12560417 TI - Cancer survivors in the United States: age, health, and disability. AB - BACKGROUND: Relatively little is known about the health and disability of adult cancer survivors. As a way to explore these issues, data from the National Health Interview Survey (years 1998-2000) were analyzed. METHODS: Comparisons were made between cancer survivors (n = 4878) and those without a history of cancer (n = 90,737), using both descriptive statistics and logistic-regression models on general health status, psychological disability, limitations in activities of daily living, physical function, and health-related ability to work. Among cancer survivors, health and disability status were assessed by cancer site or type, age at diagnosis, and years since cancer diagnosis. RESULTS: Compared with individuals without a history of cancer or other chronic disease, cancer survivors without other chronic diseases were significantly more likely to report being in fair or poor health (odds ratio, or OR, 2.97), a psychological disability (OR 2.18), limitations of activities of daily living or instrumental activities of daily living (OR 2.22), functional limitations (OR 1.74), and, among those under the age of 65, being unable to work because of a health condition (OR 3.22). The likelihood of poor health and disability was much higher among cancer survivors who also reported comorbid chronic conditions. CONCLUSIONS: Providers caring for cancer survivors should be made aware of the long-term health consequences of cancer and consider appropriate supportive care for their patients. The identification of long-term effects of cancer that contribute to disability and the interventions needed to ameliorate these and their consequences should become a more prominent aspect of the research agenda. PMID- 12560418 TI - Commentary on Dr. Kane's article. The future history of geriatrics: geriatrics at the crossroads. PMID- 12560419 TI - The future history of geriatrics: consulting the experts. PMID- 12560420 TI - Unrecognized parkinsonism in acute care medical patients receiving neurological consultations. PMID- 12560421 TI - Acute illnesses, ADL decline, and hospitalization. PMID- 12560422 TI - Taking stock of volume-outcome studies. PMID- 12560423 TI - Molecular diagnostics: assays, tissues, progress, and pitfalls. PMID- 12560424 TI - Prognosis after rectal cancer treatment in blacks and whites: advanced stage at diagnosis or other factors? PMID- 12560425 TI - neu/erbB-2 overexpression and response to hormonal therapy in premenopausal women in the adjuvant breast cancer setting: will it play in Peoria? part II. PMID- 12560426 TI - Role of surgeon volume in radical prostatectomy outcomes. AB - PURPOSE: To examine the effect of hospital and surgeon volume on postoperative outcomes and to determine whether hospital or surgeon volume is the stronger predictor. PATIENTS AND METHODS: Using 1997 to 1998 claims data from a national 5% random sample of Medicare beneficiaries, we identified 2,292 men who underwent radical prostatectomy at 1,210 hospitals by 1,788 surgeons. Hospitals were classified as high (> or = 60 per year) or low (< 60 per year) volume according to radical prostatectomy experience over the 2-year period. Surgeons were classified as high (> or = 40 per year) or low (< 40 per year) volume. Multivariate logistic regression was performed to control for patient demographics and comorbidities when assessing the association of hospital and surgeon volume with in-hospital complications, length of stay, and anastomotic stricture rates. In-hospital complications included cardiac, respiratory, vascular, wound, genitourinary, and miscellaneous surgical and medical conditions. RESULTS: High-volume surgeons had half the complication risk (odds ratio [OR] = 0.53; 95% confidence interval [CI], 0.32 to 0.89) and shorter lengths of stay (4.1 v 5.2 days, P =.03) compared with low-volume surgeons. High volume hospital patients tended to have fewer anastomotic strictures (OR = 0.72; 95% CI, 0.49 to 1.04). Patient age (> or = 75 years) was associated with more complications (OR = 1.9; 95% CI, 1.39 to 2.70), more anastomotic strictures (OR = 2.2; 95% CI, 1.54 to 3.15), and longer hospital stays (parameter estimate = 2.26; 95% CI, 1.75 to 2.77). CONCLUSION: Surgeon volume is inversely related to in hospital complications and length of stay in men undergoing radical prostatectomy. Hospital volume is not significantly associated with outcomes after adjusting for physician volume. Further study is necessary to elucidate the mechanism of the volume-outcome effect. PMID- 12560427 TI - Thymidylate synthase expression in hepatic tumors is a predictor of survival and progression in patients with resectable metastatic colorectal cancer. AB - PURPOSE: To investigate the role of thymidylate synthase (TS),p53, and epidermal growth factor receptor (EGF-R) expressions in hepatic tumors in predicting overall survival (OS), progression-free survival (PFS), and hepatic progression free survival (HPFS) in patients with resectable metastatic colorectal cancer who were randomly assigned to receive either systemic chemotherapy (SYS) alone or systemic and hepatic arterial infusion (HAI+SYS) chemotherapy following liver surgery. PATIENTS AND METHODS: Tissues from metastatic tumors were collected during liver resection from 156 patients, and marker expressions were determined using immunohistochemistry on frozen samples. Univariate associations between marker expressions and baseline variables with OS, PFS, and HPFS were examined. Independent predictors of outcome were determined using a multivariate Cox model. RESULTS: In multivariate analyses, TS overexpression was found to be an independent factor of poor prognosis in OS (P <.01), PFS (P =.06), and HPFS (P <.01). In addition, resection margin was a significant independent factor for all three outcomes. Patients who received HAI+SYS experienced delayed progression in general, and in the liver, specifically. Increased levels of serum alkaline phosphatase correlated with hepatic progression. We also found a significant TS treatment interaction for OS (P =.01) in multivariate analysis. In particular, TS+ patients receiving HAI+SYS had significantly higher survival than those receiving SYS (64 month sv 21 months; P =.01). CONCLUSION: TS levels in hepatic tumors and resection margin are independent predictors of survival and progression in patients with metastatic colorectal cancer, whereas p53 and EGFR are not independent predictors. Treatment with HAI + SYS significantly improved the survival profile of TS+ patients. PMID- 12560428 TI - Prognosis after rectal cancer in blacks and whites participating in adjuvant therapy randomized trials. AB - PURPOSE: National health statistics indicate that blacks have lower survival rates from colorectal cancer than do whites. This disparity has been attributed to differences in stage at diagnosis and other disease features, extent and quality of treatment, and socioeconomic factors. We evaluated outcomes for blacks and whites with rectal cancer who participated in randomized clinical trials of the National Surgical Adjuvant Breast and Bowel Project (NSABP). The randomized trial setting enhances uniformity in disease stage and treatment plan among all participants. PATIENTS AND METHODS: The study included black (N = 104) or white (N = 1,070) patients from two serially conducted NSABP randomized trials for operable rectal cancer. Recurrence-free survival and survival were compared using statistical modeling to account for differences in patient and disease characteristics between the groups. RESULTS: Blacks and whites had largely similar disease features at diagnosis. After adjustment for patient and tumor prognostic covariates, the black/white recurrence hazard ratio (HR) was 1.25 (95% confidence interval [CI], 0.94 to 1.66). The mortality HR was somewhat larger at 1.45 (95% CI = 1.09 to 1.93). Outcomes were improved for both groups in the more recent trial, which employed systemic adjuvant chemotherapy in all treatment arms. CONCLUSION: Recurrence-free survival was modestly less favorable for blacks, whereas overall survival was more disparate. Outcomes between groups were more comparable than those noted in national health statistics surveys and other studies. Adequate treatment access and the identification of new prognostic factors that can identify patients at high risk of recurrence are needed to ensure optimal outcomes for rectal cancer patients of all racial/ethnic backgrounds. PMID- 12560429 TI - Phase II trial of systemic continuous fluorouracil and subcutaneous recombinant interferon Alfa-2b for treatment of hepatocellular carcinoma. AB - PURPOSE: Because cirrhosis is extremely common in hepatocellular carcinoma (HCC) in the United States, and it precludes the use of several chemotherapy agents, this phase II trial of fluorouracil (FU) and recombinant interferon alfa-2b (rIFNalpha2b) in HCC was launched with the assumption that it could be tolerated by cirrhotics. PATIENTS AND METHODS: Forty-three patients with HCC (34), and fibrolamellar HCC (FLHCC; nine) were treated with continuous intravenous (IV) FU (200 mg/m2/d x 21 every 28 days) and subcutaneous (SC) rIFNalpha2b (4 million U/m2) three times weekly. Survival was determined in all 43 patients, and response could be assessed in 28 HCC and 8 FLHCC patients. RESULTS: The median ages of the patients were 63.5 and 19 years among HCC and FLHCC patients, respectively. Liver cirrhosis was present among 71% of HCC patients but among none of the FLHCC patients. Nine of 36 (25%; four of 28 [14%] HCC patients; five of eight [62.5%] FLHCC patients) patients in which a response could be assessed had a complete response (CR; one patient with FLHCC and no patients with HCC) or partial response (PR; eight patients [four HCC and four FLHCC patients]). Four HCC patients underwent resection, and two had a histologic CR; one HCC patient with a PR underwent orthotopic liver transplantation. One FLHCC patient also underwent resection without clear margins. Overall median survival was 19.5 months (95% confidence interval [CI], 11.2 to 27.8 months); median survival was 15.5 months (95% CI, 8.5 to 22.5 months) among HCC patients, and that of FLHCC patients was 23.1 months (95% CI, 10.3 to 35.9 months). Overall grade 3 or 4 toxicity included stomatitis (32.6%), fatigue (4.7%), and hematologic toxicity (9.3%). CONCLUSION: Continuous IV FU and thrice-weekly SC rIFNalpha2b are an effective treatment, especially for FLHCC, and may have a neoadjuvant role in this disease. This regimen has activity in HCC and can be tolerated even by cirrhotic patients. PMID- 12560430 TI - Whole body 18FDG-PET and the response of esophageal cancer to induction therapy: results of a prospective trial. AB - PURPOSE: Whole-body 18F-fluorodeoxyglucose ([18F]FDG) positron emission tomography (PET) imaging before and after induction therapy was prospectively evaluated in patients with esophageal cancer to determine whether changes in PET images could measure response to therapy. PATIENTS AND METHODS: Between April 1997 and April 1999, 39 patients (34 men and five women; median age, 59 years; range, 36 to 76 years) with esophageal cancer were prospectively enrolled in a single-institution clinical trial of staging, including PET, induction therapy, restaging including PET, and esophagectomy. All patients undergoing esophagectomy after induction therapy (n = 17) were followed either to recurrence, to death, or through a disease-free interval of at least 24 months. RESULTS: PET after standard staging studies and before therapy imaged undetected sites of metastatic disease in six patients (15%). Restaging (including PET) after induction therapy did not identify any patients with disease progression or any patients with loco regionally unresectable disease at exploration. The median decrease in the standardized uptake value (SUV) during induction therapy was 59%. After R0 esophagectomy, the 2-year disease-free and overall survival was 38% and 63%, respectively, among patients who had a less than 60% decrease in SUV, and 67% and 89%, respectively, among patients who had a greater than 60% decrease in SUV (P =.055 and P =.088, respectively). CONCLUSION: Compared with conventional imaging, PET detects additional sites of metastatic disease at initial evaluation. After induction therapy, PET did not add to the estimation of loco-regional resectability and did not detect new distant metastases. However, changes in [18F]FDG PET may predict disease-free and overall survival after induction therapy and resection in patients with esophageal cancer. Further evaluation in larger trials is warranted. PMID- 12560431 TI - Impact of diabetes mellitus on outcomes in patients with colon cancer. AB - PURPOSE: To determine the influence of diabetes mellitus on long-term outcomes and treatment-related toxicity among patients with curatively resected colon cancer. PATIENTS AND METHODS: This study was a cohort study within a large, randomized adjuvant chemotherapy trial of 3,759 patients with high-risk stage II and stage III colon cancer treated between 1988 and 1992 throughout the United States. In the cohort, 287 patients were identified as having diabetes mellitus. With a median follow-up of 9.4 years, we analyzed differences in overall survival (OS) and colon cancer recurrence as well as treatment-related toxicity between patients with diabetes and those without diabetes. RESULTS: At 5 years, patients with diabetes mellitus, compared with patients without diabetes, experienced a significantly worse disease-free survival (DFS; 48% diabetics v 59% nondiabetics; P <.0001), OS (57% v 66%; P <.0001), and recurrence-free survival (RFS; 56% v 64%, P =.012). Median survival was 6.0 years and 11.3 years for diabetics and nondiabetics, respectively. Compared with patients without a history of diabetes, those with diabetes had a 42% increased risk of death from any cause (P <.0001) and 21% increased risk for recurrence (P =.05) after adjustment for other predictors of colon cancer outcome. Treatment-related toxicities were similar between the two groups, although patients with diabetes experienced an increase in treatment-related diarrhea. CONCLUSION: Patients with diabetes mellitus and high-risk stage II and stage III colon cancer experienced a significantly higher rate of overall mortality and cancer recurrence, even after adjustment for other predictors of colon cancer outcome. These results underscore the need for further research to understand the mechanism that underlies this relation. PMID- 12560432 TI - Estrogen receptor classification for hepatocellular carcinoma: comparison with clinical staging systems. AB - PURPOSE: Several scoring systems to evaluate patients with hepatocellular carcinoma (HCC) exist. A good scoring system should provide information on prognosis and guide therapeutic decisions. The presence of variant liver estrogen receptor (ER) transcripts in the tumor has been shown to be the strongest negative predictor of survival in HCC. The aim of this study was to compare the predictive value of the commonly applied clinical scoring systems for survival of patients with HCC with that of the evaluation of ER in patients with HCC (molecular scoring system). MATERIALS AND METHODS: HCC was staged according to the Okuda classification, Barcelona Clinic Liver Cancer classification, Italian classification system (CLIP), French classification, and ER status in 96 patients. Analysis of survival was performed according to the Kaplan-Maier test and was made for each classification system and ER. A comparison between classifications was made by univariate and multivariate analysis. RESULTS: Among the clinical classification systems, only the CLIP was able to identify patient populations with good, intermediate, and poor prognosis. On multivariate analysis, ER classification was shown to be the best predictive classification for survival of patients with HCC (P <.0001). This difference was the result of a better allocation of patients with ominous prognosis (variant ER) having nevertheless good clinical score. CONCLUSION: The evaluation of the presence of wild-type or variant ER transcripts in the tumor is the best predictor of survival in patients with HCC. Its accuracy in discriminating patients with good or unfavorable prognosis is significantly greater than that of the commonly used scoring systems for the staging of HCC. PMID- 12560433 TI - Elevated expression of valosin-containing protein (p97) in hepatocellular carcinoma is correlated with increased incidence of tumor recurrence. AB - PURPOSE: Valosin-containing protein (VCP; also known as p97) has been shown to be associated with antiapoptotic function and metastasis via activation of the nuclear factor-kappaB signaling pathway. In this study, association of VCP expression with recurrence of hepatocellular carcinoma (HCC) and patient survival was examined. PATIENTS AND METHODS: VCP expression in 170 patients (139 male and 31 female) with ages ranging from 31 to 81 years (median, 61 years) was analyzed by quantitative reverse-transcription polymerase chain reaction (RT-PCR) and immunohistochemistry, in which staining intensity in tumor cells was categorized as weaker (level 1) or equal to or stronger (level 2) than that in endothelial cells. RESULTS: Immunohistochemically, 57 patients (35.2%) showed level 1, and 105 patients (64.8%) showed level 2, VCP expression. Quantitative RT-PCR analysis revealed higher VCP mRNA expression in level 2 patients (n = 7) than level 1 (n = 4) (P <.05). Patients with VCP-level 2 HCC showed higher rate of portal vein invasion in the tumor (P <.01) and poorer disease-free and overall survival (P <.0001 and P <.05, respectively) compared with level 1 patients. Multivariate analysis revealed VCP expression level, tumor multiplicity, and degree of fibrosis in the noncancerous liver tissue to be independent prognosticators for disease-free and overall survival. VCP level was an indicator for disease-free survival in each early- (I and II) and advanced- (III and IV) stage group of pathologic tumor-node-metastasis classification (P <.001 and P <.01, respectively). CONCLUSION: VCP expression level has prognostic significance for disease-free and overall survival of patients with HCC. PMID- 12560434 TI - Her-2/neu overexpression and response to oophorectomy plus tamoxifen adjuvant therapy in estrogen receptor-positive premenopausal women with operable breast cancer. AB - PURPOSE: Studies evaluating the relationship of HER-2/neu breast tumor status and response to adjuvant endocrine therapy have reached conflicting conclusions about resistance of HER-2/neu-positive tumors to this treatment. We studied 282 patients participating in a randomized controlled trial of adjuvant oophorectomy and tamoxifen or observation who had estrogen receptor-positive tumors and whose tumors were evaluated for HER-2/neu overexpression by immunohistochemistry. PATIENTS AND METHODS: Univariate and multivariate Cox proportional hazards regression models and Kaplan-Meier disease-free and overall survival estimate methods were used. RESULTS: HER-2/neu overexpression was a negative prognostic factor for overall survival. In univariate analyses, in HER-2/neu-positive patients, the hazard ratio (HR) for disease-free survival (DFS) with adjuvant endocrine therapy was 0.37 (95% confidence interval [CI], 0.26 to 0.89); for HER 2/neu-negative patients, the corresponding HR for DFS was 0.48 (95% CI, 0.31 to 0.71). The overall survival (OS) data were HR=0.26 (95% CI, 0.07 to 0.92) and HR=0.68 (95% CI, 0.32 to 1.42) for HER-2/neu-positive and HER-2/neu-negative patients, respectively. In multivariate models, the P values for tests of interaction of HER-2/neu status and response to adjuvant endocrine therapy were 0.18 and 0.07 for DFS and OS, respectively. Kaplan-Meier DFS and OS curves and 3 year DFS estimates were consistent in showing greater benefit to the HER-2/neu positive subgroup given adjuvant treatment. CONCLUSION: HER-2/neu overexpression does not adversely and may favorably influence response to adjuvant oophorectomy and tamoxifen treatment in patients with estrogen receptor-positive tumors. PMID- 12560435 TI - HER2 overexpression and doxorubicin in adjuvant chemotherapy for resectable breast cancer. AB - PURPOSE: Human epidermal growth factor receptor 2 (HER2) overexpression was found to predict a good response in breast carcinoma patients treated with doxorubicin (Adriamycin [ADM]). Evidence from our recent study indicates that node-positive patients respond to cyclophosphamide, methotrexate, and fluorouracil (CMF) regardless of HER2 status. We address the issue of whether therapy regimens including CMF and ADM versus CMF alone have the same therapeutic effect in patients with HER2+ and HER2- tumors in terms of relapse-free survival (RFS) and overall survival (OS). METHODS: Archival specimens of the primary tumors from 506 patients in a prospective clinical trial were stained with the anti-HER2 monoclonal antibody CB11. Originally, patients were randomly allocated to receive either 12 courses of intravenous CMF or eight courses of the same regimen followed by four cycles of ADM. RFS and OS were analyzed by a Cox model taking into account treatment, HER2 status, and the interaction between treatment and HER2 status, adjusting for the effect of other known clinical and biopathologic factors. RESULTS: Analysis of survival rates indicates a possible differential effect of treatment in the patients grouped according to HER2 status. Improved RFS and OS were observed in the HER2+ subgroup after treatment with CMF plus ADM versus CMF alone. With a median follow-up of 15 years, the hazard ratio (HR) for RFS was 0.83 in HER2+ tumors and 1.22 in HER2- tumors. The effect of treatment was more evident on OS in HER2+ patients (HR = 0.61; CI, 0.32 to 1.16) than in HER2- patients (HR = 1.26). CONCLUSION: Our data indicate that adding ADM to CMF might be beneficial for patients with HER2+ tumors. PMID- 12560436 TI - Effect of upper extremity exercise on secondary lymphedema in breast cancer patients: a pilot study. AB - PURPOSE: To examine the effect of a progressive upper-body exercise program on lymphedema secondary to breast cancer treatment. METHODS: Fourteen breast cancer survivors with unilateral upper extremity lymphedema were randomly assigned to an exercise (n = 7) or control group (n = 7). The exercise group followed a progressive, 8-week upper-body exercise program consisting of resistance training plus aerobic exercise using a Monark Rehab Trainer arm ergometer. Lymphedema was assessed by arm circumference and measurement of arm volume by water displacement. Patients were evaluated on five occasions over the experimental period. The Medical Outcomes Trust Short-Form 36 Survey was used to measure quality of life before and after the intervention. Significance was set at alpha < or = 0.01. RESULTS: No changes were found in arm circumference or arm volume as a result of the exercise program. Three of the quality-of-life domains showed trends toward increases in the exercise group: physical functioning (P =.050), general health (P =.048), and vitality (P =.023). Mental health increased, although not significantly, for all subjects (P =.019). Arm volume measured by water displacement was correlated with calculated arm volume (r =.973, P <.001), although the exercise and control group means were significantly different (P <.001). CONCLUSIONS: Participation in an upper-body exercise program caused no changes in arm circumference or arm volume in women with lymphedema after breast cancer, and they may have experienced an increase in quality of life. Additional studies should be done in this area to determine the optimum training program. PMID- 12560437 TI - Diminished survival in patients with inner versus outer quadrant breast cancers. AB - PURPOSE: To evaluate the role of breast tumor location on survival in patients with breast cancer. PATIENTS AND METHODS: We evaluated tumor location within the breast on breast cancer-specific survival (BCSS) and overall survival (OS) in patients with invasive breast cancer using the Surveillance, Epidemiology, and End-Results (SEER) registries in the United States. Effects on survival were evaluated according to age, stage, tumor site, tumor size, grade, axillary lymph node status, extent of surgery, and radiotherapy (RT). A multivariate model was used with complete data on 45,880 patients. The median follow-up was 59 months. RESULTS: Patients with outer tumor location demonstrated superior BCSS on Kaplan Meier analysis for both local stage (node-negative, P <.001) and regional stage disease (node-positive, P =.0002). For BCSS, the hazard ratio (HR) for inner quadrant location compared with outer quadrant was 1.31 (95% confidence interval [CI], 1.19 to 1.37; P <.001); and for OS, the HR was 1.12 (95% CI, 1.05 to 1.17; P <.001). When ER and PR status were included in the model, the HR for inner quadrant location compared with outer quadrant was 1.27 for BCSS (95% CI, 1.16 to 1.40; P <.001) and 1.11 for OS (95% CI, 1.03 to 1.19; P =.004). Patients treated by lumpectomy that received RT had a superior OS compared with patients that did not receive RT in both local (HR, 0.52; 95% CI, 0.48 to 0.61; P <.001) and regional (HR, 0.63; 95% CI, 0.56 to 0.72; P <.001) stage disease. Mastectomy patients with local stage disease that received RT had a diminished OS (HR, 1.24; 95% CI, 1.02 to 1.50; P =.033). CONCLUSION: On multivariate analysis, incorporating data on age, stage, tumor site, tumor size, grade, ER and PR status, axillary lymph node status, extent of surgery, and RT, this SEER registry based study demonstrates that medial tumor location adversely impacts BCSS and OS. PMID- 12560438 TI - Carbonic anhydrase IX expression, a novel surrogate marker of tumor hypoxia, is associated with a poor prognosis in non-small-cell lung cancer. AB - PURPOSE: To evaluate carbonic anhydrase (CA) IX as a surrogate marker of hypoxia and investigate the prognostic significance of different patterns of expression in non-small-cell lung cancer (NSCLC). METHODS: Standard immunohistochemical techniques were used to study CA IX expression in 175 resected NSCLC tumors. CA IX expression was determined by Western blotting in A549 cell lines grown under normoxic and hypoxic conditions. Measurements from microvessels to CA IX positivity were obtained. RESULTS: CA IX immunostaining was detected in 81.8% of patients. Membranous (m) (P =.005), cytoplasmic (c) (P =.018), and stromal (P <.001) CA IX expression correlated with the extent of tumor necrosis (TN). The mean distance from vascular endothelium to the start of tumor cell positivity was 90 micro m, which equates to an oxygen pressure of 5.77 mmHg. The distance to blood vessels from individual tumor cells or tumor cell clusters was greater if they expressed mCA IX than if they did not (P <.001). Hypoxic exposure of A549 cells for 16 hours enhanced CA IX expression in the nuclear and cytosolic extracts. Perinuclear (p) CA IX (P =.035) was associated with a poor prognosis. In multivariate analysis, pCA IX (P =.004), stage (P =.001), platelet count (P =.011), sex (P =.027), and TN (P =.035) were independent poor prognostic factors. CONCLUSION: These results add weight to the contention that mCA IX is a marker of tumor cell hypoxia. The absence of CA IX staining close to microvessels suggests that these vessels are functionally active. pCA IX expression is representative of an aggressive phenotype. PMID- 12560439 TI - Predictors of biochemical outcome with salvage conformal radiotherapy after radical prostatectomy for prostate cancer. AB - PURPOSE: To identify predictors of biochemical outcome following radiotherapy in patients with a rising prostate-specific antigen (PSA) after radical prostatectomy for prostate cancer. PATIENTS AND METHODS: One hundred fifteen patients with a rising PSA after radical prostatectomy received salvage three dimensional conformal radiotherapy (3D-CRT) alone or with neoadjuvant androgen deprivation. Tumor-related and treatment-related factors were evaluated to identify predictors of subsequent PSA failure. RESULTS: The median follow-up time after 3D-CRT was 42 months. The 4-year actuarial PSA relapse-free survival, distant metastasis-free survival, and overall survival rates were 46%, 83%, and 95%, respectively. Multivariate analysis, which was limited to 70 patients receiving radiation without androgen deprivation therapy, showed that negative/close margins (P =.03), absence of extracapsular extension (P <.01), and presence of seminal vesicle invasion (P <.01) were independent predictors of PSA relapse after radiotherapy. Neoadjuvant androgen deprivation did not improve the 4-year PSA relapse-free survival in patients with positive margins, extracapsular extension, and no seminal vesicle invasion (P =.24). However, neoadjuvant androgen deprivation did improve PSA relapse-free survival when one or more of these variables were absent (P =.03). CONCLUSIONS: Salvage 3D-CRT can provide biochemical control in selected patients with a rising PSA after radical prostatectomy. Among patients with positive margins and no poor prognostic features, 77% achieved PSA control after salvage 3D-CRT. Salvage neoadjuvant androgen deprivation therapy may improve short-term biochemical control, but it requires further study. PMID- 12560440 TI - Prognostic significance of reverse transcriptase polymerase chain reaction for prostate-specific antigen in metastatic prostate cancer: a nested study within CALGB 9583. AB - PURPOSE: To determine whether reverse transcriptase polymerase chain reaction (RT PCR) to detect circulating prostate-specific antigen (PSA)-positive cells is a prognostic factor for survival in hormone refractory prostate cancer and to validate the prognostic importance of this test in relation to other known prognostic factors. PATIENTS AND METHODS: A single centralized laboratory received and analyzed whole blood for RT-PCR for PSA for a subset of patients enrolled on two Cancer and Leukemia Group B (CALGB) randomized trials (CALGB 9583 and CALGB 9480). Using 9583, a prognostic model was developed and an independent data set (CALGB 9480) was used to validate the fitted model. RESULTS: Of 162 patients in 9583, 91 (56%) patients were negative for RT-PCR for PSA and 71 (44%) patients were positive. The median survival time was 21 months (95% confidence interval [CI], 18 to 27 months) for RT-PCR-negative patients compared with 11 months (95% CI, 8 to 15 months) for RT-PCR-positive patients (P < or =.001). In multivariable analysis, the hazard ratio (HR) for death was 1.7 (95% CI, 1.2 to 2.4; P =.006) for positive RT-PCR patients compared with negative RT-PCR patients. A fitted model that incorporated RT-PCR for PSA and other factors was used to classify patients from 9480 into one of two risk groups: low or high. We observed good agreement between the observed and predicted survival probabilities for the two risk groups. CONCLUSION: RT-PCR to detect PSA-positive circulating cells is confirmed to be a significant prognostic factor of survival in patients with hormone refractory prostate cancer. This model could be used to stratify patients in randomized phase III trials. PMID- 12560441 TI - Management and outcome differences in supraglottic cancer between Ontario, Canada, and the Surveillance, Epidemiology, and End Results areas of the United States. AB - PURPOSE: We compared the management and outcome of supraglottic cancer in Ontario, Canada, with that in the Surveillance, Epidemiology, and End Results (SEER) Program areas in the United States. METHODS: Electronic, clinical, and hospital data were linked to cancer registry data and supplemented by chart review where necessary. Stage-stratified analyses compared initial treatment and survival in the SEER areas (n = 1,643) with a random sample from Ontario (n = 265). We also compared laryngectomy rates at 3 years in those patients 65 years and older at diagnosis. RESULTS: Radical surgery was more commonly used in SEER, with absolute differences increasing with increasing stage: I/II, 17%; III, 36%; and IV, 45%. The 5-year survival rates were 74% in Ontario and 56% in SEER for stage I/II disease (P =.01), 55.7% in Ontario and 46.8% in SEER for stage III disease (P =.40), and 28.5% in Ontario and 29.1% in SEER for stage IV disease (P =.28). Cancer-specific survival results mirrored the overall survival results with the exception of stage IV disease, for which 34.6% of Ontario patients survived their cancer compared with 38.1% in SEER (P =.10). This stage IV difference was more pronounced when we further controlled for possible cause of death errors by restricting the comparison to patients with a single primary cancer (P =.01). Three-year actuarial laryngectomy rates differed. In stage I/II, these rates were 3% in Ontario compared with 35% in SEER (P < 10(-3)). In stage III disease, the rates were 30% and 54%, respectively (P =.03), and in stage IV disease they were 33% and 64% (P =.002). CONCLUSION: There are large differences in the management of supraglottic cancer between the SEER areas of the United States and Ontario. Long-term larynx retention was higher in Ontario, where radiotherapy is widely regarded as the treatment of choice and surgery is reserved for salvage. In stages I to III, survival was similar in the two regions despite the differences in treatment policy. In stage IV, there may be a small survival advantage in the U.S. SEER areas related to the higher use of primary surgery. PMID- 12560442 TI - Matched and mismatched allogeneic stem-cell transplantation from unrelated donors using combined graft-versus-host disease prophylaxis including rabbit anti-T lymphocyte globulin. AB - PURPOSE: With improved HLA-typing techniques, it is presently unclear what degree of identity is necessary for successful unrelated-donor stem-cell transplantation (UD SCT). Here, we describe the outcome after matched and mismatched UD SCT using a graft-versus-host disease (GVHD) prophylaxis including high-dose rabbit anti-T lymphocyte globulin (ATG). PATIENTS AND METHODS: One hundred adult patients (median age, 37 years; range, 17 to 65 years) with hematologic malignancies underwent transplantation in early disease (first complete remission [CR1] or first chronic phase [CP1]; n = 34) or in advanced disease (second complete remission or second chronic phase, no remission, refractory; n = 66) with nondepleted bone marrow (n = 87) or peripheral-blood-derived (n = 13) stem cells from an HLA-A, HLA-B, HLA-DRB1*, or HLA-DQB1* identical (n = 75) or mismatched (one antigen, n = 21; two to three antigens, n = 4) unrelated donor. GVHD prophylaxis consisted of rabbit ATG before transplantation in addition to cyclosporine and short-course methotrexate. RESULTS: The cumulative incidence of acute GVHD degrees II- degrees IV was 21% (95% confidence interval [CI], 14% to 33%) and 20% (95% CI, 9% to 44%) and acute GVHD degrees III- degrees IV was 5.3% (95% CI, 2% to 14%) and 4% (95% CI, 0.6% to 28%) in HLA-matched and HLA mismatched transplantations, respectively. The risk for extensive chronic GVHD was 43% (95% CI, 32% to 59%) and 44% (95% CI, 26% to 75%) for HLA-matched and HLA mismatched patients, respectively. The risk of relapse at 4 years was 17% (95% CI, 7% to 43%) and 43% (95% CI, 31% to 60%) for CR1/CP1 and advanced disease patients, respectively. With a median follow-up of 1,068 days (range, 12 to 1,958 days), 3-year disease-free and overall survival for patients who underwent transplantation in CR1/CP1 was 63% (95% CI, 46% to 81%) and 75% (95% CI, 59% to 90%), respectively; and for patients with advanced disease, it was 34% (95% CI, 22% to 46%) and 39% (95% CI, 25% to 53%), respectively. CONCLUSION: A certain degree of one antigen mismatching may not compromise the outcome after UD SCT when using this rabbit ATG in addition to standard GVHD prophylaxis regimen. PMID- 12560443 TI - Randomized, multicenter, open-label study of pegfilgrastim compared with daily filgrastim after chemotherapy for lymphoma. AB - PURPOSE: The primary objective was to assess the duration of grade 4 neutropenia (neutrophil count < 0.5 x 10(9)/L) after one cycle of chemotherapy with etoposide, methylprednisolone, cisplatin, and cytarabine in patients randomly assigned to receive one dose of pegfilgrastim or daily filgrastim after chemotherapy. Febrile neutropenia, neutrophil profiles, time to neutrophil recovery, pharmacokinetics, and safety were also assessed. PATIENTS AND METHODS: An open-label, randomized, phase II study was designed to compare the effects of a single subcutaneous injection of pegfilgrastim (sustained-duration filgrastim) 100 micro g/kg per chemotherapy cycle (n = 33) with daily subcutaneous injections of filgrastim 5 micro g/kg (n = 33) in patients receiving salvage chemotherapy for relapsed or refractory Hodgkin's or non-Hodgkin's lymphoma. RESULTS: The incidence of grade 4 neutropenia in the pegfilgrastim and filgrastim groups was 69% and 68%, respectively. In addition, the mean duration of grade 4 neutropenia was similar in both groups (2.8 and 2.4 days, respectively). The results for the two groups were also not significantly different for febrile neutropenia, neutrophil profile, time to neutrophil recovery, or toxicity profile. A single subcutaneous injection of pegfilgrastim 100 micro g/kg produced a sustained serum concentration relative to daily subcutaneous injections of filgrastim. Filgrastim treated patients received a median of 11 injections per cycle. CONCLUSION: Pegfilgrastim was safe and well tolerated in this patient population. A single injection of pegfilgrastim per chemotherapy cycle provided neutrophil support with safety and efficacy similar to that provided by daily injections of filgrastim. Once-per-cycle administration of pegfilgrastim simplifies the management of neutropenia and may have important clinical benefits for patients and healthcare providers. PMID- 12560444 TI - Prospective evaluation of a follow-up schedule in cutaneous melanoma patients: recommendations for an effective follow-up strategy. AB - PURPOSE: To prospectively examine and evaluate the results of follow-up procedures in a large cohort of cutaneous melanoma patients. PATIENTS AND METHODS: This was a prospective study in 2,008 consecutive patients with stage I to IV cutaneous melanoma from 1996 to 1998 on the yield of stage-appropriate follow-up examinations according to the German guidelines. Documentation of patient and follow-up data comprised patient demography, primary tumor specifics, and any clinical and technical examinations performed. The detection of metastasis was classified as early or late, and the means of their detection and the resulting overall survival probabilities were examined. RESULTS: A total of 3,800 clinical examinations and 12,398 imaging techniques were documented. Sixty two second primary melanomas in 46 patients and 233 disease recurrences in 112 patients were detected during this time. In stage I to III disease, physical examination was responsible for the discovery of 50% of all recurrences. In the primary tumor stages, 21% of all recurrences were discovered by lymph node sonography, with the majority being classified as early detection. Forty-eight percent of the recurrences were classified as early detection, and these patients had a significant benefit of overall survival probability. CONCLUSION: The results of our study suggest that an elaborated follow-up schedule in cutaneous melanoma is suitable for the early detection of second primary melanomas and early recurrences. The intensity of clinical and technical examinations can be reduced during follow-up of patients in the primary tumor stages and may be intensified in locoregional disease. Recommendations for an effective follow-up strategy are outlined. PMID- 12560445 TI - Renal cell carcinoma in children: a clinicopathologic study. AB - PURPOSE: To identify the prognostic factors, treatment, and outcome of children affected by renal cell carcinoma (RCC). PATIENTS AND METHODS: The series included 41 patients (18 males and 23 females) with a median age of 124 months observed at the 11 Italian Association for Pediatric Hematology and Oncology centers from January 1973 to January 2001. Clinical data, surgical notes, pathologic findings, and summaries of therapy were taken from the charts. RESULTS: Seven (17%) of the 41 patients had a papillary histology, and 34 (82.4%) had nonpapillary histology. Eighteen patients (43.9%) had stage I, one patient (2.4%) had stage II, two patients (4.8%) had stage IIIA, 10 patients (24.3%) had stage IIIB, and nine patients (21.9%) had stage IV disease. One patient had a bilateral involvement at diagnosis. Seven patients experienced disease recurrence. Lung and liver were the most common distant lesions and usually were fatal. In this study, the major factor influencing the prognosis was the stage. Event-free survival at 20 years was 53.5% for all patients. Overall survival at 20 years was 54.9% for all patients. CONCLUSION: RCC is a rare disease in children and adolescents. This neoplasm has a different clinical presentation in children compared with adults but the same outcome. In our experience, patients with localized disease could be cured by nephrectomy alone. Prospective studies in a larger number of patients are needed to confirm radiation therapy and biologic response modifiers as effective adjunct therapy in RCC stage III. The alternative therapy seems warranted in patients with advanced disease. PMID- 12560446 TI - Value of P-glycoprotein and clinicopathologic factors as the basis for new treatment strategies in high-grade osteosarcoma of the extremities. AB - PURPOSE: To evaluate the prognostic value of P-glycoprotein and clinicopathologic parameters in a large series of high-grade osteosarcoma (OS) patients treated at the Rizzoli Institute. PATIENTS AND METHODS: With the use of immunohistochemistry, P-glycoprotein was assessed in 149 patients with primary, nonmetastatic, high-grade OS who were homogeneously treated with chemotherapy protocols based on doxorubicin, high-dose methotrexate, and cisplatin and the addition of ifosfamide in the postoperative phase. RESULTS: P-glycoprotein positivity was found in 47 of 149 cases (32%) and was significantly associated with a higher incidence of relapse and a worse outcome, as was age younger than 12 years and tumor volume greater then 150 mL at diagnosis. Multivariate analysis further confirmed the prognostic value of these parameters, which all were independent adverse prognostic factors. Event-free survival and proportional hazards regression analyses confirmed that overexpression of P-glycoprotein at clinical onset is the most important adverse prognostic factor for high-grade OS patients treated with these chemotherapy protocols. CONCLUSION: Increased P glycoprotein levels, together with tumor volume and age, should be taken into consideration to identify, at time of diagnosis, subgroups of OS patients with a higher risk of recurrence. This subgroup identification will constitute the basis for drawing individualized treatment protocols on the basis of risk evaluation, with the aim of using more aggressive chemotherapy, or combination chemotherapy with other adjuvants, only in those patients for which more aggressive regimens are strictly necessary and warranted. PMID- 12560447 TI - Dispositional optimism predicts survival status 1 year after diagnosis in head and neck cancer patients. AB - PURPOSE: The aim of this study was to investigate the hypothesis that, independent of other known prognostic factors, pessimistic head and neck (H&N) cancer patients have a greater risk of being dead 1 year after diagnosis than do optimistic patients. PATIENTS AND METHODS: A prospective observational study design was used with a cohort of H&N cancer patients diagnosed during the period from March 1, 1997, to August 31, 1998, at the Centre Hospitalier Universitaire, Clermont-Ferrand, France. Dispositional optimism (DO) was evaluated at baseline using a French version of the Life Orientation Test translated and validated for this study. One-year survival status was collected on all subjects. The analysis of the hypothesized association between DO and 1-year survival was performed using multiple logistic regression analysis, controlling for other sociodemographic and clinical variables. RESULTS: The sample size was 101 patients, representing all but one of those patients fitting the inclusion criteria who were diagnosed during the recruitment period. Of these, 51 were alive at 1 year after diagnosis, 45 were dead, and five were lost to follow-up. The multivariate analysis was performed on the data from the 96 subjects in whom 1-year survival status was known. Controlling for known predictors of H&N cancer survival, pessimistic subjects (odds ratio [OR], 1.12; 95% confidence interval [CI], 1.01 to 1.24) and those living alone (OR, 4.14; 95% CI, 1.21 to 14.17) were more likely than optimistic subjects and those living with others to be dead at 1 year. CONCLUSION: The results of this study of a cohort of French H&N cancer patients indicate that dispositional optimism predicts 1-year survival independent of other sociodemographic and clinical variables. PMID- 12560448 TI - Approaching the end of life: attitudes, preferences, and behaviors of African American and white patients and their family caregivers. AB - PURPOSE: To investigate differences in attitudes, preferences, and behaviors regarding end of life in terminally ill patients and their designated family caregivers. PATIENTS AND METHODS: 68 African-American and white patients with stage III-B or IV lung or stage IV colon cancer and 68 patient-designated family caregivers interviewed between December 1999 and May 2001. RESULTS: White patients were more likely to have a durable power of attorney (34% v 8%, P =.01) and were more likely to have a living will (LW; 41% v 11%, P =.004) than were African-American patients. More African-American than white patients desired the use of life-sustaining measures (cardiopulmonary resusitation [CPR], mechanical ventilation, tube feeding) in their current condition (all P >.12). In a near death condition, African-American patients were more likely than white patients to desire each of the life-sustaining measures (all P <.004). There was no patient-caregiver agreement beyond chance regarding preferences for initiation of CPR, tube feeding, or mechanical ventilation in the patient's current condition or in the near-death condition. In the near-death condition in patients without LWs, there was disagreement in 46% of patient-caregiver pairs about CPR, in 50% about mechanical ventilation, and in 43% about tube feeding. CONCLUSION: Although most patients and families endorse the primacy of the patient in decisions at end of life, the majority do not take supporting actions. Disagreements between patients and families about the use of life-sustaining measures in patients without LWs may result in patients' preferences being superseded at end of life. PMID- 12560449 TI - Does delay in starting treatment affect the outcomes of radiotherapy? A systematic review. AB - PURPOSE: The objective of this study was to synthesize what is known about the relationship between delay in radiotherapy (RT) and the outcomes of RT. METHODS: A systematic review of the world literature was conducted to identify studies that described the association between delay in RT and the probability of local control, metastasis, and/or survival. Studies were classified by clinical and methodologic criteria and their results were combined using a random-effects model. RESULTS: A total of 46 relevant studies involving 15,782 patients met our minimum methodologic criteria of validity; most (42) were retrospective observational studies. Thirty-nine studies described rates of local recurrence, 21 studies described rates of distant metastasis, and 19 studies described survival. The relationship between delay and the outcomes of RT had been studied in diverse situations, but most frequently in breast cancer (21 studies) and head and neck cancer (12 studies). Combined analysis showed that the 5-year local recurrence rate (LRR) was significantly higher in patients treated with adjuvant RT for breast cancer more than 8 weeks after surgery than in those treated within 8 weeks of surgery (odds ratio [OR] = 1.62, 95% confidence interval [CI], 1.21 to 2.16). Combined analysis also showed that the LRR was significantly higher among patients who received postoperative RT for head and neck cancer more than 6 weeks after surgery than among those treated within 6 weeks of surgery (OR = 2.89; 95% CI, 1.60 to 5.21). There was little evidence about the impact of delay in RT on the risk of metastases or the probability of long-term survival in any situation. CONCLUSION: Delay in the initiation of RT is associated with an increase [corrected] in LRR in breast cancer and head and neck cancer. Delays in starting RT should be as short as reasonably achievable. PMID- 12560450 TI - Uncommon presentations of non-Hodgkin's lymphoma: case 1. Intravascular large B cell lymphoma: diagnosis on prostate biopsy. PMID- 12560451 TI - Uncommon presentations of non-Hodgkin's lymphoma: case 2. Non-Hodgkin's lymphoma of the ileocecal region. PMID- 12560452 TI - Uncommon presentations of non-Hodgkin's lymphoma: case 3. Primary renal lymphoma. PMID- 12560453 TI - Setting goals to maintain hope. PMID- 12560456 TI - Demystified. Human endogenous retroviruses. AB - Human endogenous retroviruses (HERVs) are a family of viruses within our genome with similarities to present day exogenous retroviruses. HERVs have been inherited by successive generations and it is possible that some have conferred biological benefits. However, several HERVs have been implicated in certain cancers and autoimmune diseases. This article demystifies these retroviruses by providing an insight into HERVs, their means of classification, and a synopsis of HERVs implicated in cancer and autoimmunity. Furthermore, the biological roles of HERVs are explored. PMID- 12560454 TI - Molecular aspects of type 1 diabetes. AB - Type 1 diabetes is a T cell mediated autoimmune disease, characterised by the selective destruction of pancreatic beta cells, and susceptibility is determined by a combination of genetic and environmental factors. The environmental agents implicated include viruses and dietary factors, although none has yet been shown to be directly responsible for triggering beta cell autoimmunity. The genetic factors that influence disease risk have been subjected to more intensive study and two gene regions of major importance have been identified: the human leucocyte antigen locus and the insulin gene. This review will focus on the mechanisms by which these genes might influence the risk of developing type 1 diabetes. PMID- 12560457 TI - Effect of Helicobacter pylori on apoptosis and apoptosis related genes in gastric cancer cells. AB - BACKGROUND/AIMS: Helicobacter pylori induces the apoptosis of gastric epithelial cells in vivo and in vitro. However, the molecular mechanism has not been clarified. The aim of this study was to investigate the effect of H pylori on the apoptosis of gastric epithelial cells and the expression of apoptosis related genes in vitro. METHODS: Human gastric adenocarcinoma SGC-7901 cells were co cultured with a cytotoxic H pylori strain, NCTC 11637, at various densities ranging from 3.2 x 10(4) to 1.0 x 10(8) colony forming units (CFU)/ml for 48 hours. Apoptosis in gastric cells was determined by transmission electron microscopy, Hoechst 33258 fluorochrome staining, and flow cytometry. The expression of apoptosis related proteins, Bcl-2, Bax, and c-Myc, was measured by an immunohistochemical method, and c-Myc mRNA expression was determined by the reverse transcription-polymerase chain reaction. RESULTS: Helicobacter pylori induces morphological changes typical of apoptosis. Both fluorochrome staining and flow cytometry showed that the apoptotic index began to increase when H pylori were at a density of > 1.6 x 10(4) CFU/ml, and in a density dependent manner (p < 0.01; one way ANOVA). The expression of the Bax and c-Myc proteins and of c-Myc mRNA was increased, whereas Bcl-2 expression was decreased after co culture for 48 hours. CONCLUSIONS: Helicobacter pylori induced apoptosis in gastric epithelial cells is mediated by altered expression of the products of the Bcl-2, Bax, and c-Myc genes. PMID- 12560459 TI - A novel polymerase chain reaction assay to detect Mycoplasma genitalium. AB - AIMS: To design and validate a polymerase chain reaction (PCR) assay targeting the 16S rRNA gene of Mycoplasma genitalium. METHODS: Primers were designed that were complementary to the 16S rRNA gene sequence of M genitalium. After optimisation of the reaction conditions, the PCR was tested against nine M genitalium strains, a dilution series of M genitalium DNA, and a panel of common microorganisms. The PCR was also challenged in parallel with a published assay against 54 urine specimens from men with urethritis. RESULTS: The expected 341 bp product was produced on amplification of material from all M genitalium strains and from none of the other microorganisms tested. The lower limit of detection was 50 genome copies. The new assay detected M genitalium DNA in nine of 54 men with urethritis, in comparison with eight positive specimens detected with the alternative PCR. CONCLUSIONS: This novel PCR targeting the M genitalium 16S rRNA gene has been optimised and now provides a sensitive and specific alternative or addition to the available MgPa gene targeting assays. PMID- 12560460 TI - Pseudobacteraemia in a patient with neutropenic fever caused by a novel paenibacillus species: Paenibacillus hongkongensis sp. nov. AB - AIMS: To characterise a strain of Gram negative aerobic straight or slightly curved rods (HKU3) isolated from the blood culture of a 9 year old Chinese boy with neutropenic fever and pseudobacteraemia. METHODS: The isolate was phenotypically investigated by standard biochemical methods using conventional biochemical tests, scanning electron microscopy, and transmission electron microscopy. Genotypically, the 16S rRNA gene of the bacterium was amplified by the polymerase chain reaction (PCR) and sequenced. The sequence of the PCR product was compared with known 16S rRNA gene sequences in the Genbank by multiple sequence alignment. The G + C content was determined by thermal denaturation. A phylogenetic tree was constructed by the PileUp method. RESULTS: The cells of the bacterial strain were aerobic, sporulating, Gram negative straight or slight curved rods. The bacterium grew on horse blood agar as non haemolytic, grey colonies of 1 mm in diameter after 24 hours of incubation at 37 degrees C in ambient air. No enhancement of growth was seen in 5% CO(2). It grew at 50 degrees C as pinpoint colonies after 72 hours of incubation, but did not grow at 65 degrees C or on MacConkey agar. It was non-motile. It produced catalase (weakly positive) and cytochrome oxidase. It reduced nitrate, produced beta galactosidase, hydrolysed esculin, and utilised sodium acetate. A scanning electron micrograph of the bacterium showed straight or slightly curved rods. A transmission electron micrograph of the cell wall of the bacterium revealed multiple electron dense layers, including the outer membrane, middle murein layer, and inner cytoplasmic membrane, compatible with its Gram smear appearance. 16S rRNA gene sequencing showed that there were 7.7%, 8.0%, 8.2%, and 8.6% differences between the 16S rRNA gene sequence of the bacterium and those of Paenibacillus macerans, Paenibacillus borealis, Bacillus ehimensis, and Paenibacillus amylolyticus, respectively. The mean (SD) G + C content of the bacterium was 47.6 (2.1) mol%. Phylogenetically, it belongs to the genus paenibacillus (previously called group 3 bacillus). CONCLUSIONS: A bacterium that exhibited phenotypic and genotypic characteristics that are very different from closely related members of paenibacillus was the cause of pseudobacteraemia in a patient with neutropenic fever. A new species, Paenibacillus hongkongensis sp. nov. is proposed, for which HKU3 is the type strain. PMID- 12560462 TI - Helicobacter pylori genotyping in gastric adenocarcinoma and MALT lymphoma by multiplex PCR analyses of paraffin wax embedded tissues. AB - BACKGROUND: Gastric infection with Helicobacter pylori is the major cause of chronic active gastritis and is associated with the pathogenesis of peptic ulcer and gastric carcinoma. Gastric mucosal damage involves both host and H pylori dependent factors, such as the presence of the cag pathogenicity island and allelic variations of the vacA and iceA genes. AIMS: To evaluate the association of these virulence factors with the development of gastric malignancies, a retrospective study was performed on archived tissue routinely obtained for diagnostic histopathology. METHODS: DNA was extracted from formalin fixed, paraffin wax embedded gastric tissue sections of 93 patients with chronic active gastritis (n = 39), adenocarcinoma (n = 28), or mucosa associated lymphoid tissue (MALT) lymphoma (n = 24). The extracted DNA was used to perform a polymerase chain reaction based, simultaneous analysis of the following: (1) cagA status, (2) allelic variation of the iceA genes (iceA1, iceA2), allelic variation of the signal peptide (s1a, s1b, s2) and the midregion (m1, m1a, m2) of the vacA gene. RESULTS: The iceA1 gene showed a 3.6 fold and the vacA s1a variant a 4.2 fold higher prevalence in gastric adenocarcinoma than in gastritis. The combined presence of both the vacA s1a and iceA1 genes had a 5.6 fold higher frequency in adenocarcinoma. The vacA m2 allele was the predominant subtype in MALT lymphoma and the combination of the vacA m2 subtypes with the vacA s1 and the iceA1 variants occurred in MALT lymphoma nearly five times more often than in chronic active gastritis. CONCLUSIONS: Certain H pylori subtype combinations possess a differentiating and predictive value for the development of gastric adenocarcinoma and MALT lymphoma. PMID- 12560463 TI - Accurate molecular detection of melanoma nodal metastases: an assessment of multimarker assay specificity, sensitivity, and detection rate. AB - BACKGROUND: The application of lymphoscintigraphy followed by sentinel lymph node (SN) biopsy to patients with primary melanoma has revolutionised the ability to identify accurately, yet conservatively, those patients who harbour occult nodal metastases. The molecular detection of SN micrometastases facilitates the cost effective analysis of the entire SN using multiple markers. Currently, a lack of marker specificity is the main barrier preventing the molecular evaluation of SN tissue from becoming clinically applicable. AIMS: To develop a reproducible multimarker reverse transcription-polymerase chain reaction (RT-PCR) assay, with the emphasis on achieving high specificity for the accurate detection of melanoma metastases in nodal tissue. METHODS: Three pigment cell specific (PCS) markers tyrosinase, Pmel-17, and MART-1-and one cancer testis antigen (CTA)-MAGE-3-were selected for use in a multimarker RT-PCR assay. The conditions for this assay were optimised. RESULTS: High specificity was achievable for each marker by optimising the PCR cycle number such that unwanted transcripts (that is, illegitimate transcripts and/or specific transcripts from other low abundance nodal cell types) remained undetectable in appropriate controls (normal visceral nodes). Tyrosinase was 100% specific at 40 PCR cycles, MAGE-3 and MART-1 at 35 PCR cycles, and Pmel-17 at 30 PCR cycles. Tyrosinase proved to be the most sensitive marker, detecting 10 melanoma cells in 0.1 g of nodal tissue. CONCLUSIONS: Excellent reproducibility of the entire nodal processing and RT-PCR protocol for the detection of very low numbers of melanoma cells in nodal tissue was shown, although there is a risk of false positives using the PCS markers alone, because of an approximate 4-8.5% incidence rate of nodal nevi in melanoma draining SNs (these nevi being absent in all other normal nodes). MAGE-3 was shown to be the only marker that is not expressed by melanocytes. However, because not all melanomas express MAGE-3, it is recommended that more emphasis should be placed on the development of a panel of CTA markers to ensure a zero false positive rate and to provide optimum detection. PMID- 12560466 TI - p73 gene mutations in gastric adenocarcinomas. AB - BACKGROUND/AIMS: The p73 gene encodes a protein that shares structural and functional homology with the p53 gene product. The highest degree of homology is in the DNA binding domain, which is the region of p53 that is most frequently mutated in cancer. In contrast to p53 there is little evidence that p73 acts as a classic tumour suppressor gene. Because of the similarities between the p53 and p73 genes and the high frequency of mutation of p53, this study was designed to investigate the p73 gene in patients with gastric adenocarcinoma. METHODS: The mutational status of the p73 gene was investigated in a series of 13 cases of gastric adenocarcinoma from the antro-pyloric region and the gastro-oesophageal junction, using the polymerase chain reaction, single strand conformational polymorphism, and direct DNA sequencing. RESULTS: A glutamine to arginine mutation was detected in exon 5 of the p73 gene in a case of adenocarcinoma at the gastro-oesophageal junction. CONCLUSION: Although limited to a small series of cases, these results suggest that p73 may have a potential pathogenetic role in this tumour. PMID- 12560464 TI - Salivary gland expression of transforming growth factor beta isoforms in Sjogren's syndrome and benign lymphoepithelial lesions. AB - AIM: Transforming growth factor beta (TGF-beta) is involved in the control of autoimmune reactions, cell proliferation, and the accumulation of lymphocytes within organs. The aim of this study was to determine the expression of TGF-beta in salivary glands from patients with primary Sjogren's syndrome (SS) and benign lymphoepithelial lesions (BLEL) with emphasis on ductal epithelium. METHODS: Immunoperoxidase staining for TGF-beta isoforms and Ki67 antigen was performed on formalin fixed sections of labial glands from patients with primary SS (n = 15) and controls (n = 5) and parotid glands reported as BLEL (n = 5) or normal (n = 5). Ductal expression of TGF-beta was quantified by absorbance measurements using image analysis. The specificity of staining was confirmed by peptide blocking studies. RESULTS: All TGF-beta isoforms were detected within the cytoplasm of most lymphocytes, endothelial cells, and ducts in all specimens. Acinar expression was variable and weaker than that seen in ducts. Absorbance measurements revealed that the expression of all isoforms was greater in ducts within primary SS glands than in control glands. Ductal expression in control parotid glands was greater than that seen in BLEL glands, irrespective of the presence of adjacent lymphoid infiltrates. Comparisons between control specimens showed that ductal expression of all isoforms was highest in parotid glands, whereas no differences were detected between primary SS and BLEL glands. Ki67 positive lymphocytes and duct cells were mainly restricted to pathological specimens, with BLEL glands containing larger populations of positive cells than primary SS glands. CONCLUSION: These results demonstrate complex and variable changes in ductal expression of TGF-beta in primary SS and BLEL, which may be important in the control of lymphoid infiltration and the proliferation of lymphocytes and ductal epithelium. PMID- 12560467 TI - Molecular determination of carriage of the mecA locus in coagulase negative staphylococci in screening swabs from patients in an intensive care unit. PMID- 12560470 TI - Taming Winfree turbulence of scroll waves in excitable media. AB - Winfree turbulence of scroll waves is a special kind of spatiotemporal chaos that exists exclusively in three-dimensional excitable media and is currently considered one of the principal mechanisms of cardiac fibrillation. A chaotic wave pattern develops through the negative-tension instability of vortex filaments, which tend to spontaneously stretch, bend, loop, and produce an expanding tangle that fills up the volume. We demonstrate that such turbulence can readily be controlled by weak nonresonant modulation of the medium excitability. Depending on the forcing frequency, either suppression or induction of turbulence can be achieved. PMID- 12560471 TI - Spontaneous cell polarization through actomyosin-based delivery of the Cdc42 GTPase. AB - Cell polarization can occur in the absence of any spatial cues. To investigate the mechanism of spontaneous cell polarization, we used an assay in yeast where expression of an activated form of Cdc42, a Rho-type guanosine triphosphatase (GTPase) required for cell polarization, could generate cell polarity without any recourse to a preestablished physical cue. The polar distribution of Cdc42 in this assay required targeted secretion directed by the actin cytoskeleton. A mathematical simulation showed that a stable polarity axis could be generated through a positive feedback loop in which a stochastic increase in the local concentration of activated Cdc42 on the plasma membrane enhanced the probability of actin polymerization and increased the probability of further Cdc42 accumulation to that site. PMID- 12560472 TI - Tadpole skin dies autonomously in response to thyroid hormone at metamorphosis. AB - Transgenic tadpoles that express a dominant negative thyroid hormone (TH) receptor specifically in their skin undergo normal metamorphosis, with one exception: they retain a larval epidermis over the developing adult epithelium. TH-induced death of the tadpole epidermis is inhibited by the dominant negative TH receptor whereas the TH-induced response of the neighboring fibroblasts and the cells that form the adult skin occur normally. Therefore death of the tadpole skin is a direct and cell autonomous target of TH, and its protection has no detectable influence on TH-induced changes of other cell types. PMID- 12560473 TI - Regulation of TRP channels via lipid second messengers. AB - In Drosophila photoreceptors, the light-sensitive current is mediated downstream of phospholipase C by TRP (transient receptor potential) channels. Recent evidence suggests that Drosophila TRP channels are activated by diacylglycerol (DAG) or its metabolites (polyunsaturated fatty acids), possibly in combination with the reduction in phosphatidyl inositol 4,5 bisphosphate (PIP2). Consistent with this view, diacylglycerol kinase is identified as a key enzyme required for response termination. Signaling is critically dependent upon efficient PIP2 synthesis; mutants of this pathway in combination with genetically targeted PIP2 reporters provide unique insights into the kinetics and regulation of PIP2 turnover. Recent evidence indicates that a growing number of mammalian TRP homologues are also regulated by lipid messengers, including DAG, arachidonic acid, and PIP2. PMID- 12560474 TI - The pre-ribosomal network. AB - Recent achievements in yeast functional proteomics have significantly advanced our knowledge about ribosome biogenesis. Here, we present a program developed to integrate data from various proteome analyses with cell biological data on components present in the ribosome producing factories. This program allows users to attribute factors to certain complexes and to specific steps of ribosome biogenesis. Thus, it helps to gain novel insights into the complex network involved in maturation of ribosomal subunits. The database can be accessed at the URL http://www.pre-ribosome.de. PMID- 12560476 TI - Site-specific protein modification to identify the MutL interface of MutH. AB - We have mapped the region for the protein interaction site of the Escherichia coli mismatch repair protein MutH for its activator protein MutL by a site specific protein modification approach. For this purpose we generated a cysteine free variant of MutH and 12 variants thereof, each containing a single cysteine residue at surface positions selected on the basis of available structural and sequence information for MutH. All MutH variants displayed wild type activity both in vivo and in vitro. These variants were then site-specifically modified at their cysteine residues with thiol-specific reagents and then tested for their ability to be stimulated in their DNA cleavage activity by the activator protein MutL. Thereby we were able to identify a defined region in the MutH protein that is important for interaction with MutL, and most likely represents the MutL binding site of MutH. PMID- 12560475 TI - Conservation of structure and function among tyrosine recombinases: homology based modeling of the lambda integrase core-binding domain. AB - Tyrosine recombinases participate in diverse biological processes by catalyzing recombination between specific DNA sites. Although a conserved protein fold has been described for the catalytic (CAT) domains of five recombinases, structural relationships between their core-binding (CB) domains remain unclear. Despite differences in the specificity and affinity of core-type DNA recognition, a conserved binding mechanism is suggested by the shared two-domain motif in crystal structure models of the recombinases Cre, XerD and Flp. We have found additional evidence for conservation of the CB domain fold. Comparison of XerD and Cre crystal structures showed that their CB domains are closely related; the three central alpha-helices of these domains are superposable to within 1.44 A. A structure-based multiple sequence alignment containing 25 diverse CB domain sequences provided evidence for widespread conservation of both structural and functional elements in this fold. Based upon the Cre and XerD crystal structures, we employed homology modeling to construct a three-dimensional structure for the lambda integrase CB domain. The model provides a conceptual framework within which many previously identified, functionally important amino acid residues were investigated. In addition, the model predicts new residues that may participate in core-type DNA binding or dimerization, thereby providing hypotheses for future genetic and biochemical experiments. PMID- 12560477 TI - Genetic requirements for the targeted integration of Agrobacterium T-DNA in Saccharomyces cerevisiae. AB - Agrobacterium tumefaciens delivers transferred DNA (T-DNA) into cells of plants and yeast. In plants, the T-DNA integrates at random positions into the genome by non-homologous recombination (NHR), whereas in yeast the T-DNA preferably integrates by homologous recombination (HR). Here we show that T-DNA integration by HR in yeast requires the recombination/repair proteins Rad51 and Rad52, but not Rad50, Mre11, Xrs2, Yku70 and Lig4. In the HR events a remarkable shift from insertion-type events to replacement events was observed in rad50, mre11 and xrs2 mutants. Residual integration in the rad51 mutant occurred predominantly by HR, whereas in the rad52 mutant integration occurred exclusively by NHR. Previously, we found that T-DNA integration by NHR is abolished in a yku70 mutant. Thus, Rad52 and Yku70 are the key regulators of T-DNA integration, channeling integration into either the HR or NHR pathway. PMID- 12560478 TI - Selective inhibition of transcription of the Ets2 gene in prostate cancer cells by a triplex-forming oligonucleotide. AB - The transcription factor Ets2 has a role in cancer development and represents an attractive therapeutic target. In this study, we designed a triplex-forming oligonucleotide (TFO) directed to a homopurine:homopyrimidine sequence in the Ets2 promoter. Transcription factors of the Sp family bound to this sequence and mutation of the Sp1 site reduced Ets2 promoter activity. The Ets2-TFO had high binding affinity for the target sequence and inhibited binding of Sp1/Sp3 to the overlapping site. This effect occurred with a high degree of sequence specificity. Mismatched oligonucleotides did not inhibit Sp1/Sp3 binding and mutations in the target sequence that abolished triplex formation prevented inhibition of Sp1/Sp3 binding by the TFO. The Ets2-TFO inhibited Ets2 promoter activity and expression of the endogenous gene in prostate cancer cells at nanomolar concentrations. The TFO did not affect reporter constructs with mutations in the TFO binding site and promoters of non-targeted genes. Expression of non-targeted genes was also not affected in TFO-treated cells. Collectively, these data demonstrated that the anti-transcriptional activity of the Ets2-TFO was sequence- and target-specific, and ruled out alternative, non-triplex mediated mechanisms of action. This anti-transcriptional approach may be useful to examine the effects of selective downregulation of Ets2 expression and may have therapeutic applications. PMID- 12560479 TI - Crystal structure of the complementary quadruplex formed by d(GCATGCT) at atomic resolution. AB - Here we report the crystal structure of the DNA heptanucleotide sequence d(GCATGCT) determined to a resolution of 1.1 A. The sequence folds into a complementary loop structure generating several unusual base pairings and is stabilised through cobalt hexammine and highly defined water sites. The single stranded loop is bound together through the G(N2)-C(O2) intra-strand H-bonds for the available G/C residues, which form further Watson-Crick pairings to a complementary sequence, through 2-fold symmetry, generating a pair of non-planar quadruplexes at the heart of the structure. Further, four adenine residues stack in pairs at one end, H-bonding through their N7-N6 positions, and are additionally stabilised through two highly conserved water positions at the structural terminus. This conformation is achieved through the rotation of the central thymine base at the pinnacle of the loop structure, where it stacks with an adjacent thymine residue within the lattice. The crystal packing yields two halved biological units, each related across a 2-fold symmetry axis spanning a cobalt hexammine residue between them, which stabilises the quadruplex structure through H-bonds to the phosphate oxygens and localised hydration. PMID- 12560480 TI - Does the HIV-1 primer activation signal interact with tRNA3(Lys) during the initiation of reverse transcription? AB - Reverse transcription of HIV-1 RNA is primed by a tRNA3(Lys) molecule bound at the primer binding site (PBS). Complex intermolecular interactions were proposed between tRNA3(Lys) and the RNA of the HIV-1 Mal isolate. Recently, an alternative interaction was proposed between the TPsiC stem of tRNA3(Lys) and a primer activation signal (PAS) of the Lai and Hxb2 RNAs, suggesting major structural variations in the reverse transcription complex of different HIV-1 strains. Here, we analyzed mutants of the Hxb2 RNA that prevent the interaction between the PAS and tRNA3(Lys) or/and a complementary sequence in the viral RNA. We compared the kinetics of reverse transcription of the wild type and mutant Hxb2 RNAs, using either tRNA3(Lys) or an 18mer oligoribonucleotide complementary to the PBS, which cannot interact with the PAS, as primers. We also used chemical probing to test the structure of the mutant and wild type RNAs, as well as the complex formed between the later RNA and tRNA3(Lys). These experiments, together with the analysis of long term replication data of mutant viruses obtained by C. Morrow and coworkers (Birmingham, USA) that use alternate tRNAs as primers, strongly suggest that the interaction between the Hxb2 PAS and tRNA3(Lys) does not exist. Instead, the effects of the vRNA mutations on reverse transcription seem to be linked to incorrect folding of the mutant RNAs. PMID- 12560481 TI - Analysis of Vir protein translocation from Agrobacterium tumefaciens using Saccharomyces cerevisiae as a model: evidence for transport of a novel effector protein VirE3. AB - Agrobacterium tumefaciens causes crown gall disease on a variety of plants. During the infection process Agrobacterium transfers a nucleoprotein complex, the VirD2 T-complex, and at least two Vir proteins, VirE2 and VirF, into the plant cell via the VirB/VirD4 type IV secretion system. Recently, we found that T-DNA could also be transferred from Agrobacterium to Saccharomyces cerevisiae. Here, we describe a novel method to also detect trans-kingdom Vir protein transfer from Agrobacterium to yeast, using the Cre/lox system. Protein fusions between Cre and VirE2 or VirF were expressed in AGROBACTERIUM: Transfer of the Cre-Vir fusion proteins from Agrobacterium to yeast was monitored by a selectable excision event resulting from site-specific recombination mediated by Cre on a lox-flanked transgene in yeast. The VirE2 and VirF proteins were transported to yeast via the virB-encoded transfer system in the presence of coupling factor VirD4, analogous to translocation into plant cells. The yeast system therefore provides a suitable and fast model system to study basic aspects of trans-kingdom protein transport from Agrobacterium into host cells. Using this method we showed that VirE2 and VirF protein transfer was inhibited by the presence of the Osa protein. Besides, we found evidence for a novel third effector protein, VirE3, which has a similar C-terminal signature to VirE2 and VirF. PMID- 12560482 TI - Binding of L7Ae protein to the K-turn of archaeal snoRNAs: a shared RNA binding motif for C/D and H/ACA box snoRNAs in Archaea. AB - Small nucleolar RNAs (designated as snoRNAs in Eukarya or sRNAs in Archaea) can be grouped into H/ACA or C/D box snoRNA (sRNA) subclasses. In Eukarya, H/ACA snoRNAs assemble into a ribonucleoprotein (RNP) complex comprising four proteins: Cbf5p, Gar1p, Nop10p and Nhp2p. A homolog for the Nhp2p protein has not been identified within archaeal H/ACA RNPs thus far, while potential orthologs have been identified for the other three proteins. Nhp2p is related, particularly in the middle portion of the protein sequence, to the archaeal ribosomal protein and C/D box protein L7Ae. This finding suggests that L7Ae may be able to substitute for the Nhp2p protein in archaeal H/ACA sRNAs. By band shift assays, we have analyzed in vitro the interaction between H/ACA box sRNAs and protein L7Ae from the archaeon Archaeoglobus fulgidus. We present evidence that L7Ae forms specific complexes with three different H/ACA sRNAs, designated as Afu-4, Afu-46 and Afu 190 with an apparent K(d) ranging from 28 to 100 nM. By chemical and enzymatic probing we show that distinct bases located within bulges or loops of H/ACA sRNAs interact with the L7Ae protein. These findings are corroborated by mutational analysis of the L7Ae binding site. Thereby, the RNA motif required for L7Ae binding exhibits a structure, designated as the K-turn, which is present in all C/D box sRNAs. We also identified four H/ACA RNAs from the archaeal species Pyrococcus which exhibit the K-turn motif at a similar position in their structure. These findings suggest a triple role for L7Ae protein in Archaea, e.g. in ribosomes as well as H/ACA and C/D box sRNP biogenesis and function by binding to the K-turn motif. PMID- 12560484 TI - Infection by Mycoplasma hyorhinis strongly enhances uptake of antisense oligonucleotides: a reassessment of receptor-mediated endocytosis in the HepG2 cell line. AB - This paper shows that the approximately 66 kDa band, previously isolated from the HepG2 cell line as an oligonucleotide (ON) plasma membrane 'receptor', is induced by Mycoplasma infection. Moreover, this band has been identified as the invariant membrane protein of Mycoplasma hyorhinis, p70, based on ribosomal DNA sequencing combined with ON ligand blotting after p70 immunoprecipitation by a monoclonal antibody. Whereas antibiotic treatment of infected HepG2 cells strongly decreased ON capture, as measured by a biochemical assay, conversely, deliberate infection of HeLa cells with M.hyorhinis dramatically promoted ON uptake but did not affect receptor-mediated endocytosis of transferrin. This was confirmed by confocal microscopy of infected HepG2 cells, which also showed an indistinguishable labelling pattern after exposure of living cells to fluorescent ON and after p70 immunolabelling in permeabilised fixed cells. We propose that ON binds to p70 on M.hyorhinis attached at the cell surface, after which the complex is internalised by 'piggy-back' endocytosis. PMID- 12560483 TI - Novel mitosis-specific phosphorylation of histone H3 at Thr11 mediated by Dlk/ZIP kinase. AB - Death-associated protein (DAP)-like kinase (Dlk), also known as Zipper interacting protein (ZIP) kinase, is a nuclear serine/threonine-specific kinase that phosphorylates core histones H3 and H4, and myosine light chain in vitro. It interacts with transcription and splicing factors as well as with pro-apoptotic protein Par-4 suggesting that it participates in multiple cellular processes. To explore the significance of histone phosphorylation by Dlk, we determined the phosphorylation site in H3 and generated phosphospecific antibodies for in vivo analyses. Interestingly, Dlk/ZIP kinase phosphorylated histone H3 at a novel site, Thr11, rather than Ser10, which is characteristic of mitotic chromosomes. Immunoblotting and confocal immunofluorescence analyses demonstrated that phosphorylation of H3 at Thr11 occurred in vivo and was restricted to mitosis as well. It was discernable from prophase to early anaphase and particularly enriched at centromeres. Strikingly, during this time interval, Dlk was associated with centromeres too, as revealed by stable expression of a green fluorescent protein (GFP)-Dlk fusion protein. These findings strongly suggest that Dlk is a centromere-specific histone kinase that might play a role in labeling centromere-specific chromatin for subsequent mitotic processes. PMID- 12560485 TI - Sir2p suppresses recombination of replication forks stalled at the replication fork barrier of ribosomal DNA in Saccharomyces cerevisiae. AB - In the ribosomal DNA (rDNA) of Saccharomyces cerevisiae replication forks progressing against transcription stall at a polar replication fork barrier (RFB) located close to and downstream of the 35S transcription unit. Forks blocked at this barrier are potentially recombinogenic. Plasmids bearing the RFB sequence in its active orientation integrated into the chromosomal rDNA in sir2 mutant cells but not in wild-type cells, indicating that the histone deacetylase silencing protein Sir2 (Sir2p), which also modulates the aging process in yeast, suppresses the recombination competence of forks blocked at the rDNA RFB. Orientation of the RFB sequence in its inactive course or its abolition by FOB1 deletion avoided plasmid integration in sir2 mutant cells, indicating that stalling of the forks in the plasmid context was required for recombination to take place. Altogether these results strongly suggest that one of the functions of Sir2p is to modulate access of the recombination machinery to the forks stalled at the rDNA RFB. PMID- 12560486 TI - DNA pairing is an important step in the process of targeted nucleotide exchange. AB - Modified single-stranded DNA oligonucleotides can direct the repair of genetic mutations in yeast, plant and mammalian cells. The mechanism by which these molecules exert their effect is being elucidated, but the first phase is likely to involve the homologous alignment of the single strand with its complementary sequence in the target gene. In this study, we establish the importance of such DNA pairing in facilitating the gene repair event. Oligonucleotide-directed repair occurs at a low frequency in an Escherichia coli strain (DH10B) lacking the RECA DNA pairing function. Repair activity can be rescued by using purified RecA protein to catalyze the assimilation of oligonucleotide vectors into a plasmid containing a mutant kanamycin resistance gene in vitro. Electroporation of the preformed complex into DH10B cells results in high levels of gene repair activity, evidenced by the appearance of kanamycin-resistant colonies. Gene repair is dependent on the formation of a double-displacement loop (double-D loop), a recombination intermediate containing two single-stranded oligonucleotides hybridized to opposite strands of the plasmid at the site of the point mutation. The heightened level of stability of the double-D-loop enables it to serve as an active template for the DNA repair events. The data establish DNA pairing and the formation of the double-D-loop as important first steps in the process of gene repair. PMID- 12560487 TI - Early growth response proteins (EGR) and nuclear factors of activated T cells (NFAT) form heterodimers and regulate proinflammatory cytokine gene expression. AB - Activation of transcription factors by receptor mediated signaling is an essential step for T lymphocyte effector function. Following antigenic stimulation of T cells the two central cytokines IL-2 and TNFalpha are co expressed and co-regulated. Two important transcription factors, i.e., early growth response (EGR) protein EGR-1 and nuclear factors of activated T cells (NFAT) protein NFATc, regulate transcription of the human IL-2 cytokine and the same combination of EGR and NFAT proteins seems relevant for coordinated cytokine expression. Here we demonstrate that the zinc finger protein EGR-1 and two members of the NFAT protein family bind simultaneously to adjacent elements position -168 to -150 within the TNFalpha promoter. Both promoter sites are important for TNFalpha gene transcription as shown by transfection assays having the IL-2 and TNFalpha promoters linked to a luciferase reporter. The use of promoter deletion constructs with the zinc finger protein (ZIP), the NFAT binding element or a combination of both deleted show a functional cooperation of these elements and of their binding factors. These experiments demonstrate that EGR-1 as well as EGR-4 functionally cooperate with NFAT proteins and induce expression of both cytokine genes. Using tagged NFATc and NFATp in glutathione S-transferase pull down assays showed interaction and physical complex formation of each NFAT protein with recombinant, as well as native, EGR-1 and EGR-4 proteins. Thus EGR NFAT interaction and complex formation seems essential for human cytokine expression as adjacent ZIP and NFAT elements are conserved in the IL-2 and TNFalpha gene promoters. Binding of regulatory EGR and NFAT factors to these sites and the functional interaction and formation of stable heterodimeric complexes indicate an important role of these factors for gene transcription. PMID- 12560489 TI - New role of zCRY and zPER2 as regulators of sub-cellular distributions of zCLOCK and zBMAL proteins. AB - The core oscillator that generates circadian rhythm in eukaryotes consists of transcription/translation-based autoregulatory feedback loops by which clock gene products negatively regulate their own expression. Control of the accumulation and nuclear entry of the negative regulators PER and CRY is believed to be a key step in these loops. We clarified the mutual interaction between zebrafish clock related proteins and their sub-cellular localizations in NIH3T3 cells. Six CRYs exist in zebrafish, of which zCRY1a strongly represses zCLOCK1: zBMAL3-mediated transcription, but zCRY3 does not. We show that zCRY1a interacts with zCLOCK1 and zBMAL3, facilitating nuclear accumulation, whereas zCRY3 associates with neither one and does not influence their sub-cellular distributions. We cloned zPer2 cDNA and showed that the protein product encoded by the cDNA acts as a moderate transcriptional repressor. In our sub-cellular localization studies we also found that zPER2 interacts with the zCLOCK1:zBMAL3 heterodimer, causing its cytoplasmic retention. zCRY1a and zPER2 apparently have opposite effects on the sub-cellular distribution of zCLOCK:zBMAL heterodimer. We speculate that the opposite regulation of the sub-cellular distribution of this is associated with the different transcriptional repression abilities of zCRY1a and zPER2. zCRY1a acts as a potent transcriptional inhibitor by interacting directly with the zCLOCK:zBMAL heterodimer in the nucleus, whereas zPER2 maintains the zCLOCK:zBMAL heterodimer in the cytoplasm, resulting in transactivation repression. PMID- 12560488 TI - Cloning of human centromeres by transformation-associated recombination in yeast and generation of functional human artificial chromosomes. AB - Human centromeres remain poorly characterized regions of the human genome despite their importance for the maintenance of chromosomes. In part this is due to the difficulty of cloning of highly repetitive DNA fragments and distinguishing chromosome-specific clones in a genomic library. In this work we report the highly selective isolation of human centromeric DNA using transformation associated recombination (TAR) cloning. A TAR vector with alphoid DNA monomers as targeting sequences was used to isolate large centromeric regions of human chromosomes 2, 5, 8, 11, 15, 19, 21 and 22 from human cells as well as monochromosomal hybrid cells. The alphoid DNA array was also isolated from the 12 Mb human mini-chromosome DeltaYq74 that contained the minimum amount of alphoid DNA required for proper chromosome segregation. Preliminary results of the structural analyses of different centromeres are reported in this paper. The ability of the cloned human centromeric regions to support human artificial chromosome (HAC) formation was assessed by transfection into human HT1080 cells. Centromeric clones from DeltaYq74 did not support the formation of HACs, indicating that the requirements for the existence of a functional centromere on an endogenous chromosome and those for forming a de novo centromere may be distinct. A construct with an alphoid DNA array from chromosome 22 with no detectable CENP-B motifs formed mitotically stable HACs in the absence of drug selection without detectable acquisition of host DNAs. In summary, our results demonstrated that TAR cloning is a useful tool for investigating human centromere organization and the structural requirements for formation of HAC vectors that might have a potential for therapeutic applications. PMID- 12560490 TI - Improving the performance of DomainParser for structural domain partition using neural network. AB - Structural domains are considered as the basic units of protein folding, evolution, function and design. Automatic decomposition of protein structures into structural domains, though after many years of investigation, remains a challenging and unsolved problem. Manual inspection still plays a key role in domain decomposition of a protein structure. We have previously developed a computer program, DomainParser, using network flow algorithms. The algorithm partitions a protein structure into domains accurately when the number of domains to be partitioned is known. However the performance drops when this number is unclear (the overall performance is 74.5% over a set of 1317 protein chains). Through utilization of various types of structural information including hydrophobic moment profile, we have developed an effective method for assessing the most probable number of domains a structure may have. The core of this method is a neural network, which is trained to discriminate correctly partitioned domains from incorrectly partitioned domains. When compared with the manual decomposition results given in the SCOP database, our new algorithm achieves higher decomposition accuracy (81.9%) on the same data set. PMID- 12560491 TI - In vivo tumor growth inhibition and biodistribution studies of locked nucleic acid (LNA) antisense oligonucleotides. AB - Locked nucleic acids (LNA) are novel high-affinity DNA analogs that can be used as genotype-specific drugs. The LNA oligonucleotides (LNA PO ODNs) are very stable in vitro and in vivo without the need for a phosphorothiolated backbone. In this study we tested the biological fate and the efficacy in tumor growth inhibition of antisense oligonucleotides directed against the gene of the large subunit of RNA polymerase II (POLR2A) that are completely synthesized as LNA containing diester backbones. These full LNA oligonucleotides strongly reduce POLR2A protein levels. Full LNA PO ODNs appeared to be very stable compounds when injected into the circulation of mice. Full LNA PO ODNs were continuously administered for 14 days to tumor-bearing nude mice. Tumor growth was inhibited sequence specifically at dosages from 1 mg/kg/day. LNA PO ODNs appeared to be non toxic at dosages <5 mg/kg/day. Biodistribution studies showed the kidneys to have the highest uptake of LNA PO ODNs and urinary secretion as the major route of clearance. This report shows that LNA PO ODNs are potent genotype-specific drugs that can inhibit tumor growth in vivo. PMID- 12560492 TI - Functional consequences of mutations in the conserved SF2 motifs and post translational phosphorylation of the CSB protein. AB - The rare inherited human genetic disorder Cockayne syndrome (CS) is characterized by developmental abnormalities, UV sensitivity and premature aging. The cellular and molecular phenotypes of CS include increased sensitivity to UV-induced and oxidative DNA lesions. Two genes are involved: CSA and CSB. The CS group B (CSB) protein has roles in transcription, transcription-coupled repair, and base excision repair. It is a DNA stimulated ATPase and remodels chromatin in vitro. Here, we have analyzed wild-type (wt) and motif II, V and VI mutant CSB proteins. We find that the mutant proteins display different degrees of ATPase activity deficiency, and in contrast to the in vivo complementation studies, the motif II mutant is more defective than motif V and VI CSB mutants. Furthermore, CSB wt ATPase activity was studied with different biologically important DNA cofactors: DNA with different secondary structures and damaged DNA. The results indicate that the state of DNA secondary structure affects the level of CSB ATPase activity. We find that the CSB protein is phosphorylated in untreated cells and that UV irradiation leads to its dephosphorylation. Importantly, dephosphorylation of the protein in vitro results in increased ATPase activity of the protein, suggesting that the activity of the CSB protein is subject to phosphorylation control in vivo. These observations may have significant implications for the function of CSB in vivo. PMID- 12560493 TI - Taq DNA polymerase slippage mutation rates measured by PCR and quasi-likelihood analysis: (CA/GT)n and (A/T)n microsatellites. AB - During microsatellite polymerase chain reaction (PCR), insertion-deletion mutations produce stutter products differing from the original template by multiples of the repeat unit length. We analyzed the PCR slippage products of (CA)n and (A)n tracts cloned in a pUC18 vector. Repeat numbers varied from two to 14 (CA)n and four to 12 (A)n. Data was generated on approximately 10 single molecules for each clone type using two rounds of nested PCR. The size and peak areas of the products were obtained by capillary electrophoresis. A quasi- likelihood approach to the analysis of the data estimated the mutation rate/repeat/PCR cycle. The rate for (CA)n tracts was 3.6 x 10(-3) with contractions 14 times greater than expansions. For (A)n tracts the rate was 1.5 x 10(-2) and contractions outnumbered expansions by 5-fold. The threshold for detecting 'stutter' products was computed to be four repeats for (CA)n and eight repeats for (A)n or approximately 8 bp in both cases. A comparison was made between the computationally and experimentally derived threshold values. The threshold and expansion to contraction ratios are explained on the basis of the active site structure of Taq DNA polymerase and models of the energetics of slippage events, respectively. PMID- 12560494 TI - siRNAs generated by recombinant human Dicer induce specific and significant but target site-independent gene silencing in human cells. AB - RNA interference has emerged as a powerful tool for the silencing of gene expression in animals and plants. It was reported recently that 21 nt synthetic small interfering RNAs (siRNAs) specifically suppressed the expression of endogenous genes in several lines of mammalian cells. However, the efficacy of siRNAs is dependent on the presence of a specific target site within the target mRNA and it remains very difficult to predict the best or most effective target site. In this study, we demonstrate that siRNAs that have been generated in vitro by recombinant human Dicer (re-hDicer) significantly suppress not only the exogenous expression of a puromycin-resistance gene but also the endogenous expression of H-ras, c-jun and c-fos. In our system, selection of a target site is not necessary in the design of siRNAs. However, it is important to avoid homologous sequences within a target mRNA in a given protein family. Our diced siRNA system should be a powerful tool for the inactivation of genes in mammalian cells. PMID- 12560495 TI - A mini-RNA containing the tetraloop, wobble-pair and loop E motifs of the central conserved region of potato spindle tuber viroid is processed into a minicircle. AB - A Mini-RNA from potato spindle tuber viroid (PSTVd) was constructed specifically for cleavage and ligation to circles in vitro. It contains the C-domain with the so-called central conserved region (CCR) of PSTVd with a 17 nt duplication in the upper strand and hairpin structures at the left and rights ends of the secondary structure. The CCR was previously shown to be essential for processing of in vitro transcripts. When folded under conditions which favor formation of a kinetically controlled conformation and incubated in a potato nuclear extract, the Mini-RNA is cleaved correctly at the 5'- and the 3'-end and ligated to a circle. Thus, the CCR obviously contains all structural and functional requirements for correct processing and therefore may be regarded as 'processing domain' of PSTVd. Using the Mini-RNA as a model substrate, the structural and functional relevance of its conserved non-canonical motifs GAAA tetraloop, loop E and G:U wobble base pair were studied by mutational analysis. It was found that (i) the conserved GAAA tetraloop is essential for processing by favoring the kinetically controlled conformation, (ii) a G:U wobble base pair at the 5' cleavage site contributes to its correct recognition and (iii) an unpaired nucleotide in loop E, which is different from the corresponding nucleotide in the conserved loop E motif, is essential for ligation of the 5'- with the 3'-end. Hence all three structural motifs are functional elements for processing in a potato nuclear extract. PMID- 12560496 TI - The FCP1 phosphatase interacts with RNA polymerase II and with MEP50 a component of the methylosome complex involved in the assembly of snRNP. AB - RNA polymerase II transcription is associated with cyclic phosphorylation of the C-terminal domain (CTD) of the large subunit of RNA polymerase II. To date, FCP1 is the only specific CTD phosphatase, which is required for general transcription and cell viability. To identify FCP1-associated proteins, we constructed a human cell line expressing epitope-tagged FCP1. In addition to RAP74, a previously identified FCP1 interacting factor, we determined that FCP1-affinity purified extracts contain RNAPII that has either a hyper- or a hypo-phosphorylated CTD. In addition, by mass spectrometry of affinity purified FCP1-associated factors, we identified a novel FCP1-interacting protein, named MEP50, a recently described component of the methylosome complex that binds to the snRNP's Sm proteins. We found that FCP1 specifically interacts with components of the spliceosomal U small nuclear ribonucleoproteins. These results suggest a putative role of FCP1 CTD-phosphatase in linking the transcription elongation with the splicing process. PMID- 12560497 TI - Stimulation of D-loop formation by polypurine/polypyrimidine sequences. AB - Most of the approaches used to correct gene mutations in mammalian cells involve the targeting of short nucleotide molecules to homologous chromosomal sequences and the replacement of resident sequences via homologous recombination and mismatch repair. The limited efficiency and inconsistent reproducibility of these techniques are major constraints to their use in gene therapy. One of the main problems is that it is impossible to obtain reproducible results when the targeted gene loci differ. We investigated the effects of flanking sequences on homologous recombination by means of an in vitro assay of the efficiency of oligonucleotide targeting to its homologous sequence on a large duplex molecule in a reaction catalysed by the Escherichia coli RecA protein. We demonstrated that polypurine.polypyrimidine tracts (PPTs) in duplex DNA strongly stimulate the formation of D-loops with short oligodeoxynucleotides. This result was reproduced with various PPT sequences and oligonucleotides. The stimulatory effect was observed at loci as far as 4000 bp from the PPT. The formation of complexes between the oligonucleotide and the duplex molecule depended on the extent of sequence similarity between the two DNAs and the presence of the RecA protein. The stimulatory effect was inhibited by excess RecA and restored by adding heterologous DNA. We suggest that PPT sequences induce conformational changes in duplex DNA, leading to the aggregation of molecules, facilitating homology searches. We compared, in vivo, the efficiency of the oligonucleotide-mediated correction of a URA3 chromosomal mutation for sequences with and without a PPT sequence in the vicinity. Consistent with our in vitro results, the efficiency of correction was eight times higher in the presence of the PPT sequence. PMID- 12560499 TI - Ribonuclease inhibitor as an intracellular sentry. AB - Onconase (ONC) is a homolog of RNase A that is in clinical trials as a cancer chemotherapeutic agent. The toxicity of ONC and RNase A variants relies on their ability to evade the cytosolic ribonuclease inhibitor protein (RI) and degrade cellular RNA. We find that these ribonucleases are more toxic for more rapidly growing cells. The enhanced cytotoxicity does not arise from variation in the endogenous level of RI, which is virtually constant. Overproduction of RI diminishes the potency of toxic RNase A variants, but has no effect on the cytotoxicity of ONC. Thus, RI constrains the cytotoxicity of RNase A. These data provide new insights for the development of an optimal ribonuclease-based cancer chemotherapy. PMID- 12560498 TI - Genome instability in rad54 mutants of Saccharomyces cerevisiae. AB - The RAD54 gene of Saccharomyces cerevisiae encodes a conserved dsDNA-dependent ATPase of the Swi2/Snf2 family with a specialized function during recombinational DNA repair. Here we analyzed the consequences of the loss of Rad54 function in vegetative (mitotic) cells. Mutants in RAD54 exhibited drastically reduced rates of spontaneous intragenic recombination but were proficient for spontaneous intergenic recombinant formation. The intergenic recombinants likely arose by a RAD54-independent pathway of break-induced replication. Significantly increased rates of spontaneous chromosome loss for diploid rad54/rad54 cells were identified in several independent assays. Inter estingly, the increase in chromosome loss appeared to depend on the presence of a homolog. In addition, the rate of complex genetic events involving chromosome loss were drastically increased in diploid rad54/rad54 cells. Together, these data suggest a role for Rad54 protein in the repair of spontaneous damage, where in the absence of Rad54 protein, homologous recombination is initiated but not properly terminated, leading to misrepair and chromosome loss. PMID- 12560500 TI - Identification of pseudogenes in the Drosophila melanogaster genome. AB - Pseudogenes are copies of genes that cannot produce a protein. They can be detected from disruptions to their apparent coding sequence, caused by frameshifts and premature stop codons. They are classed as either processed pseudogenes (made by reverse transcription from an mRNA) or duplicated pseudogenes, arising from duplication in the genomic DNA and subsequent disablement. Historically, there is anecdotal evidence that the fruit fly (Drosophila melanogaster) has few pseudogenes. Investigators have linked this to a high deletion rate of genomic DNA, for which there is evidence from genetic experiments on genome size. Here, we apply a homology-based pipeline that was developed previously to identify pseudogenes in other eukaryotic genomes, to the fruit fly, so as to derive the first complete survey of its pseudogene population. We find approximately 100 pseudogenes, with at least a sixth of these as candidate processed pseudogenes. This gives a much lower proportion of pseudogenes (compared with the size of the proteome) than in the genomes of other eukaryotes for which data are available (human, nematode and budding yeast). Closest matching proteins to Drosophila pseudogenes are significantly longer than the average protein in its proteome (up to approximately 60% more than the average protein's length), in contrast to the situation in the three other eukaryotic genomes. This may be due to the persistence of fragments of longer genes. In the fly pseudogene population, we found most pseudogenes for serine proteases (which are more abundant in the Drosophila lineage compared with the other eukaryotes), immunoglobulin-motif-containing proteins and cytochromes P450. Data on the sequences and positions of the putative pseudogenes are available at: http://www.pseudogene.org/fly. The detection of a small number of pseudogenes in the Drosophila genome and the higher mean length for the closest matching proteins to pseudogenes (possibly because remnants of genes encoding longer proteins are more likely to persist) are further evidence for a high deletion rate of genomic DNA in the fruit fly. The data are useful for molecular evolution study in Drosophila. PMID- 12560501 TI - Synapsis and strand exchange in the resolution and DNA inversion reactions catalysed by the beta recombinase. AB - In the presence of a sequence-independent chromatin-associated protein, such as Hbsu or HMGB, the beta recombinase catalyses resolution between two directly oriented recombination sites (six sites) and both resolution and DNA inversion between two inversely oriented six sites. Assembly of the synaptic complex requires binding of the beta recombinase to the six sites and the presence of Hbsu. Whether resolution or inversion will take place depends on the relative orientation of the two six sites, the level of DNA supercoiling and the amounts of Hbsu. In this work, the topologies of the products of the resolution and inversion reactions were analysed. The resolution reaction generated mainly singly catenated DNA circles, while DNA inversion gave rise to unknotted circles and small amounts of DNA molecules containing 3- or 5-noded knots. In spite of the distinctive features of the beta system, the topology of synapsis and strand exchange during the resolution reaction is similar to that of Tn3 and gammadelta resolvases. The ability of the beta recombinase to catalyse both inversion and resolution reactions probably reflects different possible architectures of the synaptic complex, which to a large extent depends on Hbsu. PMID- 12560502 TI - Stability of 2'-deoxyxanthosine in DNA. AB - The deamination of nucleobases in DNA occurs by a variety of mechanisms and results in the formation of hypoxanthine from adenine, uracil from cytosine, and xanthine and oxanine from guanine. 2'-Deoxyxanthosine (dX) has been assumed to be an unstable lesion in cells, yet no study has been performed under biological conditions. We now report that dX is a relatively stable lesion at pH 7, 37 degrees C and 110 mM ionic strength, with a half-life (t(1/2)) of 2.4 years in double-stranded DNA. The stability of dX as a 2'-deoxynucleoside (t(1/2) = 3.7 min at pH 2; 1104 h at pH 6) was increased substantially upon incorporation into a single-stranded oligodeoxynucleotide, in which the half-life of dX at different pH values was found to range from 7.7 h at pH 2 to 17 700 h at pH 7. Incorporation of dX into a double-stranded oligodeoxynucleotide resulted in a statistically insignificant increase in the half-life to 20 900 h at pH 7. Data for the pH dependence of the stability of dX in single-stranded DNA were used to determine the rate constants for the acid-catalyzed (2.6 x 10(-5) x s(-1)) and pH independent (1.4 x 10(-8) x s(-1)) depurination reactions for dX as well as the dissociation constant for the N7 position of dX (6.1 x 10(-4) M). We conclude that dX is a relatively stable lesion that could play a role in deamination induced mutagenesis. PMID- 12560503 TI - Mismatch repair activity in mammalian mitochondria. AB - Mitochondrial DNA (mtDNA) defects cause debilitating metabolic disorders for which there is no effective treatment. Patients suffering from these diseases often harbour both a wild-type and a mutated subpopulation of mtDNA, a situation termed heteroplasmy. Understanding mtDNA repair mechanisms could facilitate the development of novel therapies to combat these diseases. In particular, mismatch repair activity could potentially be used to repair pathogenic mtDNA mutations existing in the heteroplasmic state if heteroduplexes could be generated. To date, however, there has been no compelling evidence for such a repair activity in mammalian mitochondria. We now report evidence consistent with a mismatch repair capability in mammalian mitochondria that exhibits some characteristics of the nuclear pathway. A repair assay utilising a nicked heteroduplex substrate with a GT or a GG mismatch in the beta-galactosidase reporter gene was used to test the repair potential of different lysates. A low level repair activity was identified in rat liver mitochondrial lysate that showed no strand bias. The activity was mismatch-selective, bi-directional, ATP-dependent and EDTA sensitive. Western analysis using antibody to MSH2, a key nuclear mismatch repair system (MMR) protein, showed no cross-reacting species in mitochondrial lysate. A hypothesis to explain the molecular mechanism of mitochondrial MMR in the light of these observations is discussed. PMID- 12560504 TI - DNA curvature at A tracts containing a non-polar thymine mimic. AB - We report the first experimental probing of electrostatic interactions on the pyrimidine side of a bent A tract. Although the curvature of short A tracts (A4 A6) has long been studied, its physical origins remain under debate. Current hypotheses include the influence of major-groove hydrogen bonds between propeller twisted base pairs, electrostatic effects of closely associated minor-groove cations, effects of minor-groove solvation, and stacking effects at the junctions adjacent to the A tract. We investigated this problem through the substitution of thymidines in A5 tracts by difluorotoluene deoxynucleoside (F), a non-polar molecule of the same size and shape which lacks hydrogen bonding and metal-ion complexing capabilities. Ligation experiments with phased A tracts demonstrated that F substitution has asymmetric effects on the bend angle. The strongest effects occurred at the second and third thymines where curvature was reduced from 19.8 degrees to 5.3 degrees and 9.6 degrees, respectively. Moderate effects were observed with substitutions at positions 1 and 4, while substitution at position 5 had no effect on bend angle. The results support the hypothesis that highly localized electrostatic interactions are a principal cause of A-tract curvature. Furthermore, they are most consistent with the notion that local metal ion complexation at O2 of thymine is a strong component of these interactions. PMID- 12560505 TI - A novel algorithm for computational identification of contaminated EST libraries. AB - A key goal of the Human Genome Project was to understand the complete set of human proteins, the proteome. Since the genome sequence by itself is not sufficient for predicting new genes and alternative splicing events that lead to new proteins, expressed sequence tags (ESTs) are used as the primary tool for these purposes. The high prevalence of artifacts in dbEST, however, often leads to invalid predictions. Here we describe a novel method for recognizing genomic DNA contamination and other artifacts that cannot be identified using current EST cleaning techniques. Our method uses the alignment of the entire set of ESTs to the human genome to identify highly contaminated EST libraries. We discovered 53 highly contaminated libraries and a subset of 24 766 ESTs from these libraries that probably represent contamination with genomic DNA, pre-mRNA, and ESTs that span non-canonical introns. Although this is only a small fraction of the entire EST dataset, each contaminating sequence could create a spurious transcript prediction. Indeed, in the clustering and assembly tool that we used, these sequences would have caused incorrect inference of 9575 new splice variants and 6370 new genes. Conclusions based on EST analysis, including prediction of alternative splicing, should be re-evaluated in light of these results. Our method, along with the identified set of contaminated sequences, will be essential for applications that depend on large EST datasets. PMID- 12560506 TI - Biopolymer Chain Elasticity: A novel concept and a least deformation energy principle predicts backbone and overall folding of DNA TTT hairpins in agreement with NMR distances. AB - A new molecular modelling methodology is presented and shown to apply to all published solution structures of DNA hairpins with TTT in the loop. It is based on the theory of elasticity of thin rods and on the assumption that single stranded B-DNA behaves as a continuous, unshearable, unstretchable and flexible thin rod. It requires four construction steps: (i) computation of the tri dimensional trajectory of the elastic line, (ii) global deformation of single stranded helical DNA onto the elastic line, (iii) optimisation of the nucleoside rotations about the elastic line, (iv) energy minimisation to restore backbone bond lengths and bond angles. This theoretical approach called 'Biopolymer Chain Elasticity' (BCE) is capable of reproducing the tri-dimensional course of the sugar-phosphate chain and, using NMR-derived distances, of reproducing models close to published solution structures. This is shown by computing three different types of distance criteria. The natural description provided by the elastic line and by the new parameter, Omega, which corresponds to the rotation angles of nucleosides about the elastic line, offers a considerable simplification of molecular modelling of hairpin loops. They can be varied independently from each other, since the global shape of the hairpin loop is preserved in all cases. PMID- 12560507 TI - DNA tri- and tetra-loops and RNA tetra-loops hairpins fold as elastic biopolymer chains in agreement with PDB coordinates. AB - The biopolymer chain elasticity (BCE) approach and the new molecular modelling methodology presented previously are used to predict the tri- dimensional backbones of DNA and RNA hairpin loops. The structures of eight remarkably stable DNA or RNA hairpin molecules closed by a mispair, recently determined in solution by NMR and deposited in the PDB, are shown to verify the predicted trajectories by an analysis automated for large numbers of PDB conformations. They encompass: one DNA tetraloop, -GTTA-; three DNA triloops, -AAA- or -GCA-; and four RNA tetraloops, -UUCG-. Folding generates no distortions and bond lengths and bond angles of main atoms of the sugar-phosphate backbone are well restored upon energy refinement. Three different methods (superpositions, distance of main chain atoms to the elastic line and RMSd) are used to show a very good agreement between the trajectories of sugar-phosphate backbones and between entire molecules of theoretical models and of PDB conformations. The geometry of end conditions imposed by the stem is sufficient to dictate the different characteristic DNA or RNA folding shapes. The reduced angular space, consisting of the new parameter, angle Omega, together with the chi angle offers a simple, coherent and quantitative description of hairpin loops. PMID- 12560508 TI - Reciprocal regulation of expression of the human adenosine 5'-triphosphate binding cassette, sub-family A, transporter 2 (ABCA2) promoter by the early growth response-1 (EGR-1) and Sp-family transcription factors. AB - The human ABCA2 transporter gene encodes a member of a large family of ATP binding proteins that transport a variety of macromolecules across biological membranes. We have performed luciferase reporter gene assays with promoter constructs comprising the 5'-flanking region to identify cis-regulatory DNA elements and have mapped the minimal promoter region to 321 bp upstream of the translation start site. We have discovered a functional role for two GC-boxes located in the proximal promoter of the ABCA2 gene that contain overlapping sites for the EGR-1 and Sp1 transcription factors. We observed that oligonucleotides containing overlapping EGR-1/Sp1 sites bind the Sp1, Sp3 and Sp4 transcription factors. When BE(2)-M17 cells were treated with phorbol 12-myristate 13-acetate, we observed inducible expression and binding of the EGR-1 transcription factor to the two GC-boxes. Transfection of Sp1, Sp3 or Sp4 expression constructs into Drosophila S2 induced a dose-dependent increase in transcriptional activation of the ABCA2 promoter, but transfection of EGR-1 alone failed to activate transcription. When increasing amounts of EGR-1 were transfected into the BE(2) M17 neuroblastoma cells we observed a dose-dependent decrease in expression of the ABCA2 promoter, although expression of the endogenous ABCA2 gene increased following transfection of EGR-1. PMID- 12560509 TI - Small worlds in RNA structures. AB - I consider conformational spaces of tRNA(phe) defined by sets of suboptimal structures from the perspective of small-world networks. Herein, the influence of modifications on typical small-world network properties and the shape of energy landscapes is discussed. Results indicate that natural modifications influence the degree of local clustering and mean path lengths far more than random or no modifications. High frequencies in the thermodynamic ensemble coincide with high numbers of neighboring structures that one conformation can adopt by one elementary move. Conformation spaces indicate the existence of modular substructures. It can be shown that modifications leave the nature of small-world topology untouched albeit natural modifications have a reasonable enhancing and streamlining effect on the degree of clustering and therefore on the substructures of the conformational space. PMID- 12560511 TI - Efficient incorporation of CoA, NAD and FAD into RNA by in vitro transcription. AB - Protein enzymes frequently recruit small molecule coenzymes to perform a variety of biochemical reactions. While the catalytic activities of RNA have been expanding rapidly, a similar strategy for RNA to utilize coenzymes and to increase its functional capabilities has yet to be demonstrated. A general in vitro transcription procedure has been developed to efficiently prepare RNA with coenzymes CoA, NAD and FAD covalently attached to the 5' end. These adenosine containing coenzymes initiate transcription under the T7 class II promoter by T7 RNA polymerase. In addition to the three coenzymes, other adenosine-containing molecules may be incorporated into the first nucleotide position of RNA as well. This method provides easy access to CoA-, NAD- and FAD-RNA, which may find broad applications in generating coenzyme- utilizing ribozymes. In addition, both oxidized FAD and reduced NADH are highly fluorescent. NADH-RNA and FAD-RNA can therefore be used as probes for DNA/RNA detection and for structural investigation of RNA function by fluorescence spectroscopy. PMID- 12560510 TI - Single base extension (SBE) with proofreading polymerases and phosphorothioate primers: improved fidelity in single-substrate assays. AB - Model single base extension (SBE) genotyping reactions with individual deoxy-, dideoxy- and acyclonucleoside triphosphates are monitored by MALDI-TOF mass spectrometry. Three non-proofreading DNA polymerases display remarkably high misincorporation (up to 64% of correct incorporation) when extending primers with single substrates at saturating concentrations. Introduction of one phosphorothioate (PS) linkage into the primer 3' terminus reduces misincorporation by these enzymes an average 1.4-fold (range 0- to 3.5-fold) versus correct incorporation. Combined use of 3'-PS primers with strongly proofreading DNA polymerases yields order of magnitude improvements in SBE fidelity over those produced by the equivalent non-proofreading enzymes. Errors are reduced to below MALDI-TOF detectable levels in almost all cases. The Sp diastereomer of the 3'-PS primer, which can be prepared in situ by incubation with proofreading polymerase, is stable to 3'-exonuclease activity over periods longer than 16 h. Products of correct extension by T7 DNAP are retained over 30 60 min during idling turnover at a dNTP concentration of 2.5 micro M, indicating that the assay can be applied over a broad range of substrate concentrations. These results suggest that the use of PS primers and proofreading polymerases will offer a simple and cost-effective means to improve fidelity in a range of single-substrate SBE assay formats. PMID- 12560512 TI - Single primer amplification (SPA) of cDNA for microarray expression analysis. AB - The potential of expression analysis using cDNA microarrays to address complex problems in a wide variety of biological contexts is now being realised. A limiting factor in such analyses is often the amount of RNA required, usually tens of micrograms. To address this problem researchers have turned to methods of improving detection sensitivity, either through increasing fluorescent signal output per mRNA molecule or increasing the amount of target available for labelling by use of an amplification procedure. We present a novel DNA-based method in which an oligonucleotide is incorporated into the 3' end of cDNA during second-strand cDNA synthesis. This sequence provides an annealing site for a single complementary heel primer that directs Taq DNA polymerase amplification of cDNA following multiple cycles of denaturation, annealing and extension. The utility of this technique for transcriptome-wide screening of relative expression levels was compared to two alternative methodologies for production of labelled cDNA target, namely incorporation of fluorescent nucleotides by reverse transcriptase or the Klenow fragment. Labelled targets from two distinct mouse tissues, adult liver and kidney, were compared by hybridisation to a set of cDNA microarrays containing 6500 mouse cDNA probes. Here we demonstrate, through a dilution series of cDNA derived from 10 micro g of total RNA, that it is possible to produce datasets comparable to those produced with unamplified targets with the equivalent of 30 ng of total RNA. The utility of this technique for microarray analysis in cases where sample is limited is discussed. PMID- 12560513 TI - Design of artificial transcription factors to selectively regulate the pro apoptotic bax gene. AB - The tumor suppressor p53 is the most commonly mutated gene in human cancers. Active p53 is able to stimulate the transcription of a variety of genes including the pro-apoptotic gene bax, as well as p21, a cell cycle regulator. In this study we produced novel zinc finger transcription factors that would selectively increase the expression of bax, but not of other p53 targets. Reporter gene assays in p53-negative Saos-2 cells showed that the novel zinc finger proteins stimulated transcription driven by a minimal bax promoter, but not that driven by a minimal p21 promoter. Moreover, electromobility shift assays demonstrated that the novel transcription factors could bind the bax promoter sequence with high affinity and selectivity. Expression of a five zinc finger protein (5ZFAV) in COS 7 cells resulted in an increase in Bax protein levels, indicating that this novel transcription factor could act on endogenous gene expression. Expression of 5ZFAV also drastically reduced Saos-2 cell survival; this effect could be reversed by the general caspase inhibitor B-D-FMK. These data suggest that 5ZFAV is able to induce apoptosis through increased Bax expression. Further, while expression of 5ZFAV in p53-deficient Saos-2 cells reduced cell survival, there was little effect on U-2 OS cells which have wild-type p53. Thus the selective induction of the pro-apoptotic bax gene may be a valuable adjunct to cancer chemotherapy by diminishing survival of p53-deficient tumor cells. PMID- 12560514 TI - TAMS technology for simple and efficient in vitro site-directed mutagenesis and mutant screening. AB - Site-directed mutagenesis is an invaluable tool for functional studies and genetic engineering. However, most current protocols require the target DNA to be cloned into a plasmid vector before mutagenesis can be performed, and none of them are effective for multiple-site mutagenesis. We now describe a method that allows mutagenesis on any DNA template (eg. cDNA, genomic DNA and plasmid DNA), and is highly efficient for multiple-site mutagenesis (up to 100%). The technology takes advantage of the requirement that, in order for DNA polymerases to elongate, it is crucial that the 3' sequences of the primers match the template perfectly. When two outer mutagenic oligos are incorporated together with the desired mutagenic oligos into the newly synthesised mutant strand, they serve as anchors for PCR primers which have 3' sequences matching the mutated nucleotides, thus amplifying the mutant strand only. The same principle can also be used for mutant screening. PMID- 12560515 TI - An epidemic of politics. PMID- 12560517 TI - U.K. universities. Tuition scheme sparks worries. PMID- 12560516 TI - Climate change. A perfect ocean for four years of globe-girdling drought. PMID- 12560518 TI - Spain. Research chief a victim of the oil spill? PMID- 12560519 TI - Neuroscience. Minor variation in growth-factor gene impairs human memory. PMID- 12560520 TI - Metabolic engineering. Researchers create first autonomous synthetic life form. PMID- 12560521 TI - Russia. Prestigious plant institute in jeopardy. PMID- 12560522 TI - Global health. Bill Gates plans a hit list, with NIH's help. PMID- 12560523 TI - Archaeology. Impending war stokes battle over fate of Iraqi antiquities. PMID- 12560524 TI - Ecology. Conflict over cooperation. PMID- 12560525 TI - Neuroscience. Singing in the brain. PMID- 12560526 TI - Neuroscience. How to please a persnickety female. PMID- 12560527 TI - Science policy in Japan. Researchers feel shut out as council loses its clout. PMID- 12560530 TI - Joint mathematics meetings. New skating system fails virtual replay. PMID- 12560528 TI - Joint mathematics meetings. Algorithmics = has trim logic. PMID- 12560529 TI - Joint mathematics meetings. How a scribe learned math, ca. 1800 B.C. PMID- 12560531 TI - Joint mathematics meetings. Diagram masters cry 'Venn-i, vidi, vici'. PMID- 12560532 TI - Josep Pique interview. Spain's science minister sees future in telecom. PMID- 12560533 TI - Evolution. Puzzling over the origin of species in the depths of the oldest lakes. PMID- 12560534 TI - Evolution. A window on ancient Siberia. PMID- 12560535 TI - Public health. Air pollution concerns not changed by S-PLUS flaw. PMID- 12560536 TI - Analytical chemistry. How to detect weak pairs. PMID- 12560537 TI - Computer science. Rough times ahead. PMID- 12560538 TI - Atmospheric science. Deciphering the energetics of lightning. PMID- 12560539 TI - Genetics. Where do male genes live? PMID- 12560540 TI - Structural biology. Complex II is complex too. PMID- 12560541 TI - Geology. Resources of the sea floor. PMID- 12560542 TI - Long-range paternal gene flow in the southern elephant seal. PMID- 12560543 TI - Suppressing roughness of virtual times in parallel discrete-event simulations. AB - In a parallel discrete-event simulation (PDES) scheme, tasks are distributed among processing elements (PEs) whose progress is controlled by a synchronization scheme. For lattice systems with short-range interactions, the progress of the conservative PDES scheme is governed by the Kardar-Parisi-Zhang equation from the theory of nonequilibrium surface growth. Although the simulated (virtual) times of the PEs progress at a nonzero rate, their standard deviation (spread) diverges with the number of PEs, hindering efficient data collection. We show that weak random interactions among the PEs can make this spread nondivergent. The PEs then progress at a nonzero, near-uniform rate without requiring global synchronizations. PMID- 12560544 TI - A gate-controlled bidirectional spin filter using quantum coherence. AB - We demonstrate a quantum coherent electron spin filter by directly measuring the spin polarization of emitted current. The spin filter consists of an open quantum dot in an in-plane magnetic field; the in-plane field gives the two spin directions different Fermi wavelengths resulting in spin-dependent quantum interference of transport through the device. The gate voltage is used to select the preferentially transmitted spin, thus setting the polarity of the filter. This provides a fully electrical method for the creation and detection of spin polarized currents. Polarizations of emitted current as high as 70% for both spin directions (either aligned or anti-aligned with the external field) are observed. PMID- 12560545 TI - Zero-mode waveguides for single-molecule analysis at high concentrations. AB - Optical approaches for observing the dynamics of single molecules have required pico- to nanomolar concentrations of fluorophore in order to isolate individual molecules. However, many biologically relevant processes occur at micromolar ligand concentrations, necessitating a reduction in the conventional observation volume by three orders of magnitude. We show that arrays of zero-mode waveguides consisting of subwavelength holes in a metal film provide a simple and highly parallel means for studying single-molecule dynamics at micromolar concentrations with microsecond temporal resolution. We present observations of DNA polymerase activity as an example of the effectiveness of zero-mode waveguides for performing single-molecule experiments at high concentrations. PMID- 12560546 TI - Nitriding iron at lower temperatures. AB - The microstructure in the surface layer of a pure iron plate was refined at the nanometer scale by means of a surface mechanical attrition treatment that generates repetitive severe plastic deformation of the surface layer. The subsequent nitriding kinetics of the treated iron with the nanostructured surface layer were greatly enhanced, so that the nitriding temperature could be as low as 300 degrees C, which is much lower than conventional nitriding temperatures (above 500 degrees C). This enhanced processing method demonstrates the technological significance of nanomaterials in improving traditional processing techniques and provides a new approach for selective surface reactions in solids. PMID- 12560547 TI - Asynchronous climate changes in the North Atlantic and Japan during the last termination. AB - Pollen records from the annually laminated sediment sequence in Lake Suigetsu, Japan, suggest a sequence of climate changes during the Last Termination that resembles that of the North Atlantic region but with noticeable differences in timing. An interstadial interval commenced a few centuries earlier [approximately 15,000 years before the present (yr B.P.)] than the North Atlantic GI-1 (Bolling) event. Conversely, the onset of a Younger Dryas (YD)-like cold reversal (12,300 to 11,250 yr B.P.) postdated the North Atlantic GS-1 (YD) event by a few centuries. Climate in the Far East during the Last Termination reflected solar insolation changes as much as Atlantic influences. PMID- 12560548 TI - The perfect ocean for drought. AB - The 1998-2002 droughts spanning the United States, southern Europe, and Southwest Asia were linked through a common oceanic influence. Cold sea surface temperatures (SSTs) in the eastern tropical Pacific and warm SSTs in the western tropical Pacific and Indian oceans were remarkably persistent during this period. Climate models show that the climate signals forced separately by these regions acted synergistically, each contributing to widespread mid-latitude drying: an ideal scenario for spatially expansive, synchronized drought. The warmth of the Indian and west Pacific oceans was unprecedented and consistent with greenhouse gas forcing. Some implications are drawn for future drought. PMID- 12560549 TI - Energetic radiation produced during rocket-triggered lightning. AB - Using a NaI(Tl) scintillation detector designed to operate in electrically noisy environments, we observed intense bursts of energetic radiation (>> 10 kiloelectron volts) during the dart leader phase of rocket-triggered lightning, just before and possibly at the very start of 31 out of the 37 return strokes measured. The bursts had typical durations of less than 100 microseconds and deposited many tens of megaelectron volts into the detector. These results provide strong evidence that the production of runaway electrons is an important process during lightning. PMID- 12560550 TI - Architecture of succinate dehydrogenase and reactive oxygen species generation. AB - The structure of Escherichia coli succinate dehydrogenase (SQR), analogous to the mitochondrial respiratory complex II, has been determined, revealing the electron transport pathway from the electron donor, succinate, to the terminal electron acceptor, ubiquinone. It was found that the SQR redox centers are arranged in a manner that aids the prevention of reactive oxygen species (ROS) formation at the flavin adenine dinucleotide. This is likely to be the main reason SQR is expressed during aerobic respiration rather than the related enzyme fumarate reductase, which produces high levels of ROS. Furthermore, symptoms of genetic disorders associated with mitochondrial SQR mutations may be a result of ROS formation resulting from impaired electron transport in the enzyme. PMID- 12560551 TI - Dissecting apicoplast targeting in the malaria parasite Plasmodium falciparum. AB - Transit peptides mediate protein targeting into plastids and are only poorly understood. We extracted amino acid features from transit peptides that target proteins to the relict plastid (apicoplast) of malaria parasites. Based on these amino acid characteristics, we identified 466 putative apicoplast proteins in the Plasmodium falciparum genome. Altering the specific charge characteristics in a model transit peptide by site-directed mutagenesis severely disrupted organellar targeting in vivo. Similarly, putative Hsp70 (DnaK) binding sites present in the transit peptide proved to be important for correct targeting. PMID- 12560552 TI - Spongiform degeneration in mahoganoid mutant mice. AB - mahoganoid is a mouse coat-color mutation whose pigmentary phenotype and genetic interactions resemble those of Attractin (Atrn). Atrn mutations also cause spongiform neurodegeneration. Here, we show that a null mutation for mahoganoid causes a similar age-dependent neuropathology that includes many features of prion diseases but without accumulation of protease-resistant prion protein. The gene mutated in mahoganoid encodes a RING-containing protein with E3 ubiquitin ligase activity in vitro. Similarities in phenotype, expression, and genetic interactions suggest that mahoganoid and Atrn genes are part of a conserved pathway for regulated protein turnover whose function is essential for neuronal viability. PMID- 12560553 TI - Prevention of transthyretin amyloid disease by changing protein misfolding energetics. AB - Genetic evidence suggests that inhibition of amyloid fibril formation by small molecules should be effective against amyloid diseases. Known amyloid inhibitors appear to function by shifting the aggregation equilibrium away from the amyloid state. Here, we describe a series of transthyretin amyloidosis inhibitors that functioned by increasing the kinetic barrier associated with misfolding, preventing amyloidogenesis by stabilizing the native state. The trans-suppressor mutation, threonine 119 --> methionine 119, which is known to ameliorate familial amyloid disease, also functioned through kinetic stabilization, implying that this small-molecule strategy should be effective in treating amyloid diseases. PMID- 12560554 TI - Modulation of heterochromatin protein 1 dynamics in primary Mammalian cells. AB - Heterochromatin protein 1 (HP1beta), a key component of condensed DNA, is strongly implicated in gene silencing and centromeric cohesion. Heterochromatin has been considered a static structure, stabilizing crucial aspects of nuclear organization and prohibiting access to transcription factors. We demonstrate here, by fluorescence recovery after photobleaching, that a green fluorescent protein-HP1beta fusion protein is highly mobile within both the euchromatin and heterochromatin of ex vivo resting murine T cells. Moreover, T cell activation greatly increased this mobility, indicating that such a process may facilitate (hetero)chromatin remodeling and permit access of epigenetic modifiers and transcription factors to the many genes that are consequently derepressed. PMID- 12560555 TI - Maintenance of stable heterochromatin domains by dynamic HP1 binding. AB - One function of heterochromatin is the epigenetic silencing by sequestration of genes into transcriptionally repressed nuclear neighborhoods. Heterochromatin protein 1 (HP1) is a major component of heterochromatin and thus is a candidate for establishing and maintaining the transcriptionally repressive heterochromatin structure. Here we demonstrate that maintenance of stable heterochromatin domains in living cells involves the transient binding and dynamic exchange of HP1 from chromatin. HP1 exchange kinetics correlate with the condensation level of chromatin and are dependent on the histone methyltransferase Suv39h. The chromodomain and the chromoshadow domain of HP1 are both required for binding to native chromatin in vivo, but they contribute differentially to binding in euchromatin and heterochromatin. These data argue against HP1 repression of transcription by formation of static, higher order oligomeric networks but support a dynamic competition model, and they demonstrate that heterochromatin is accessible to regulatory factors. PMID- 12560556 TI - Nuclear matrix localization and SUMO-1 modification of adenovirus type 5 E1b 55K protein are controlled by E4 Orf6 protein. AB - Human adenovirus serotype 5 encodes three proteins, E1b 55K, E4 Orf3 and E4 Orf6, which interact with each other and with components of the nucleus to regulate mRNA processing and export, viral DNA replication and p53-dependent apoptosis. Previous studies have shown that, during wild-type infection, 55K associates initially with structures termed ND10, which are sites of localization of the promyelocytic leukaemia protein, and then moves, dependent upon its interaction with Orf6, to the establishing virus replication centres. Absence of either Orf3 or Orf6 affects the localization of 55K and so may affect its function. In this study, the influence of Orf3 and Orf6 expression on the association of 55K with the insoluble matrix fraction of the nucleus and with ND10 particularly was examined. Overexpression of Orf6 was sufficient to block the association of 55K with this fraction, irrespective of the presence of Orf3. This effect depended upon the two proteins being able to interact. However, the association of 55K with ND10, which persists throughout infection in the absence of Orf6, required Orf3 to be present, thus distinguishing two subsets of matrix-associated 55K. A modified form of 55K, formation of which was blocked by mutating the known site of SUMO-1 attachment, was more abundant in the absence of Orf6 but unaffected by the absence of Orf3. Thus, this modification is favoured when 55K remains associated with the matrix but does not correlate with its stable association with ND10, many components of which are modified by SUMO-1. PMID- 12560557 TI - A 'first loop' linear epitope accessible on native hepatitis B surface antigen that persists in the face of 'second loop' immune escape. AB - Murine monoclonal antibodies (mAbs) were raised following immunization with native mutant hepatitis B surface antigen (HBsAg) purified from human sera. A set of antibodies binding to a linear epitope carried between residues 121 and 129 of the s region was demonstrated. These antibodies were shown by cross-competition assays to bind to a single epitope whose antigenicity was influenced by the TTP motif lying between residues 125 and 127. This first loop epitope remained accessible on the surface of HBsAg in spite of major second loop mutations abrogating the normal a conformational epitopes. The mAb and its binding region in the first loop are important diagnostically and may represent an importance immunological target, one that is stable in the face of immunologically driven escape. PMID- 12560558 TI - Purification and biochemical characterization of the E1 replication initiation protein of the cutaneous human papillomavirus type 1. AB - The E1 and E2 proteins encoded by papillomaviruses are required for viral DNA replication. Although E1 is the replication initiator protein, previous studies have shown that the full-length E1 protein binds to the origin weakly and with low sequence specificity. The E2 protein facilitates binding of the E1 protein to the origin, triggering the initiation of replication. The E1 protein contains ATPase, helicase and DNA unwinding activities. In vivo studies with mucosal human papillomavirus (HPV) types 11 and 18 have shown that while E1 is absolutely essential for replication, the E1 binding site is dispensable. However, both the E2 protein and E2 binding sites are required for their replication. In contrast to these HPVs, transient replication of HPV type 1, which infects cutaneous tissue, requires only the viral E1 protein and E1 binding site. To understand the basis for these differences, we have overexpressed and purified the HPV-1 E1 and E2 proteins and studied their biochemical properties. The purified E1 protein was shown to have an ATPase activity with a very low K(m) value, similar to that of the SV40 large T antigen. The E1 protein bound to the HPV-1 origin in the absence of the E2 protein and without the use of any cross-linking agents. Our results suggest that the ability of the HPV-1 E1 protein to initiate DNA replication in vivo in the absence of the E2 protein may be due to its stable interaction with the HPV-1 origin. PMID- 12560559 TI - Induction of humoral and cell-mediated immunity to hepatitis B surface antigen by a novel adjuvant activity of Oka varicella vaccine. AB - Oka varicella vaccine induces humoral and cell-mediated immunity to varicella zoster virus (VZV), even in immunocompromised hosts. This vaccine showed novel adjuvant activity against co-inoculated hepatitis B surface antigen (HBsAg). Either a mixed inoculation of HBsAg with heat-inactivated Oka varicella vaccine at one site or a separate inoculation of HBsAg and live vaccine at different sites induced an antibody response but failed to induce delayed type hypersensitivity (DTH) to HBsAg. In contrast, immunization of HBsAg mixed with live vaccine induced DTH and an enhanced antibody response to HBsAg. The adjuvant activity of Oka varicella vaccine was similar in terms of antibody production to that of alum adjuvant. A T helper cell-dominant immunity to VZV and HBsAg continued for 1 year. Oka varicella vaccine combined with a foreign antigen may serve as a novel polyvalent vaccine for the infectious diseases for which cell mediated immunity is beneficial. PMID- 12560560 TI - Down-regulation of MHC class I expression by equine herpesvirus-1. AB - There is good evidence that cytotoxic T lymphocytes play an important role in the clearance of equine herpesvirus-1 (EHV1) in horses. We have demonstrated that, in common with other alphaherpesviruses, EHV1 infection can lead to dramatic down regulation of MHC class I expression at the cell surface, a common strategy for pathogen evasion of the host immune response. This down-regulation is specific for MHC class I and does not reflect a general shut-off of host-cell protein synthesis. The use of monoclonal antibodies that recognize different MHC class I epitopes has demonstrated that the effect may be allele- or locus-specific. Use of the viral DNA synthesis inhibitor phosphonoacetic acid, which prevents late viral gene expression, showed that the effect is mediated by an immediate-early or early viral gene, and use of the protein translation inhibitor cycloheximide confirmed that an early gene is primarily responsible. The data indicate that EHV1 infection results in enhanced endocytosis of MHC class I from the cell surface; the only other herpesvirus reported to use this mechanism is human herpesvirus-8. Elucidation of the precise mechanisms used by EHV1 in this process and identification of the genes responsible may lead to improved vaccination strategies. PMID- 12560561 TI - Mutagenesis of a bovine herpesvirus type 1 genome cloned as an infectious bacterial artificial chromosome: analysis of glycoprotein E and G double deletion mutants. AB - The genome of bovine herpesvirus type 1 Schonboken was cloned as a bacterial artificial chromosome (BAC) by inserting mini F plasmid sequences into the glycoprotein (g) E gene. The resulting BAC clone, pBHV-1DeltagE, was transfected into bovine kidney cells and viable gE-negative BHV-1 (BHV-1DeltagE) was recovered. By RecE/T mutagenesis in Escherichia coli, the gG open reading frame was deleted from pBHV-1DeltagE. From the mutated BAC, double negative BHV 1DeltagE-gG was reconstituted and its growth properties were compared to those of rescuant viruses in which the gE gene was restored (BHV-1rev, BHV-1DeltagG). The mutant viruses did not exhibit markedly lowered virus titres. Plaque sizes of BHV 1DeltagE, BHV-1DeltagE-gG and BHV-1DeltagG, however, were reduced by 19 to 55 % compared to parental strain Schonboken or BHV-1rev. Our results suggested that gE and gG function independently from each other in cell-to-cell spread, because an additive effect on plaque formation was observed in the gE/gG double deletion mutant. PMID- 12560562 TI - Induction of HLA-G-restricted human cytomegalovirus pp65 (UL83)-specific cytotoxic T lymphocytes in HLA-G transgenic mice. AB - The non-classical major histocompatibility complex class I molecule HLA-G is expressed mainly by extravillous trophoblasts at the materno-foetal interface. HLA-G has been found to bind endogenously processed nonameric peptides but its function as a restriction element for a cytotoxic T cell response to viruses with tropism for trophoblastic cells has never been demonstrated. In this study, candidate viral peptides derived from human cytomegalovirus (HCMV) pp65 (UL83), which stabilized the HLA-G molecule on HLA-G-transfected T2 cells, were identified. The specific anti-pp65 cytotoxic T lymphocyte (CTL) response restricted by HLA-G in triple transgenic mice (HLA-G, human beta2m, human CD8alpha) was then investigated by injection of dendritic cells loaded with synthetic pp65-derived peptides or by infection with canarypox virus expressing pp65. Results showed that CTLs from HLA-G mice have the capacity to kill target cells either infected with recombinant vaccinia viruses expressing pp65 or loaded with specific pp65-derived peptides using HLA-G as an antigen-presenting molecule. It was also demonstrated that these HLA-G-restricted pp65-specific T cells are able to kill the human astrocytoma cell line U373, which was transfected with HLA-G and infected with HCMV. Moreover, using HLA-G tetramers refolded with a synthetic pp65-derived peptide, peptide-specific CD8(+) cells restricted by HLA-G have been detected in vivo. These findings provide the first evidence that HLA-G can select anti-HCMV-restricted CTLs in vivo, although the potency of this cytolytic response is limited (20-25 %). The weak HLA-G restricted anti-HCMV response is probably due to HLA-G-mediated inhibitory signals on the development of an antiviral CTL response. PMID- 12560563 TI - Physical interaction of Epstein-Barr virus (EBV) nuclear antigen leader protein (EBNA-LP) with human oestrogen-related receptor 1 (hERR1): hERR1 interacts with a conserved domain of EBNA-LP that is critical for EBV-induced B-cell immortalization. AB - Epstein-Barr virus (EBV) nuclear antigen leader protein (EBNA-LP) consists of W1W2 repeats and a unique C-terminal Y1Y2 domain and plays a critical role in EBV induced transformation. To identify the cellular proteins associating with EBNA LP, we performed a yeast two-hybrid screen using EBNA-LP cDNA containing a single W1W2 domain as bait and an EBV-transformed human peripheral blood lymphocyte cDNA library as the source of cellular genes. Our results were as follows. (i) A cDNA in the positive yeast colony was found to encode a cellular protein, human oestrogen-related receptor 1 (hERR1), which is a constitutive transcriptional activator of the various types of oestrogen response elements. (ii) A purified chimeric protein consisting of glutathione S-transferase (GST) fused to hERR1 specifically formed complexes with EBNA-LPs containing one (EBNA-LPR1), two (EBNA LPR2) or four W1W2 repeats (EBNA-LPR4) transiently expressed in COS-7 cells. Reciprocally, GST fused to EBNA-LPR1 or EBNA-LPR2 pulled down hERR1 transiently expressed in COS-7 cells. (iii) Mutational analyses of EBNA-LP revealed that the Y2 domain of EBNA-LP is responsible for the interaction with hERR1 and two leucines in the Y2 domain (Leu-78 and -82), which are conserved among a subset of primate gammaherpesviruses, are interactive sites for hERR1. So far, it has been reported that the only domain of EBNA-LP critical for EBV-induced transformation is the Y1Y2 domain. Potential roles of hERR1 in EBV-induced transformation are discussed. PMID- 12560564 TI - Auto-activation of the transforming viral interferon regulatory factor encoded by Kaposi's sarcoma-associated herpesvirus (human herpesvirus-8). AB - Kaposi's sarcoma-associated herpesvirus (KSHV)/human herpesvirus-8-encoded viral interferon regulatory factor (vIRF) transforms NIH3T3 cells, represses interferon signal transduction and regulates the expression of other KSHV genes. Here, we have shown that vIRF is a transcriptional activator and auto-activates its own expression. Ectopic expression of vIRF activated the vIRF promoter in KSHV negative 293, COS7, HeLa and BJAB cell lines in a dose-dependent fashion in a reporter assay and the expression of vIRF transcripts from endogenous viral genomes in BCBL-1 and BC-1 cells latently infected with KSHV. Deletion analysis identified two cis elements, named Vac1 and Vac2, in the vIRF promoter that were responsive to vIRF activation. vIRF auto-activation via Vac1 but not Vac2 was repressed by Tis, a transcriptional silencer in the vIRF promoter. Neither Vac1 nor Vac2 contain any interferon-stimulated response element (ISRE)-like sequences and are unresponsive to induction with interferon-beta and -gamma. These results indicate that KSHV uses the mechanism of auto-activation to regulate the expression of a viral transforming protein to efficiently evade host tumour suppressor pathways. PMID- 12560565 TI - Absence of a functional defect in CD8+ T cells during primary murine gammaherpesvirus-68 infection of I-A(b-/-) mice. AB - The murine gammaherpesvirus-68 kills I-A(b-/-) mice despite the presence of virus specific CD8+ cytotoxic T lymphocytes (CTL). This has raised the possibility that these CTL are functionally abnormal. Here, no difference was observed between I A(b-/-) mice and I-A(b+/+) controls in virus-specific CTL function, T cell receptor usage, or surface phenotype. Thus CTL immunity was independent of CD4+ T cells in a chronic herpesvirus infection, but was still inadequate to control virus replication. PMID- 12560566 TI - Early pathogenesis of Autographa californica multiple nucleopolyhedrovirus and Helicoverpa zea single nucleopolyhedrovirus in Heliothis virescens: a comparison of the 'M' and 'S' strategies for establishing fatal infection. AB - Nucleopolyhedroviruses (NPVs) (Baculoviridae) produce fatal infections in larval lepidopteran insects. NPVs are designated SNPVs or MNPVs based on whether the occlusion-derived virus (ODV) that initiates primary midgut infections contains single (S) or multiple (M) nucleocapsids. The principal consequence of this ODV packaging is that primary target cells infected with the M phenotype receive multiple nucleocapsids, whereas those infected by the S phenotype receive only one. To determine the biological significance of this difference in the initial infection strategy, a comparison of the primary and secondary infection patterns of the recombinants Helicoverpa zea SNPV (HzSNPV-hsp70/lacZ) and Autographa californica MNPV (AcMNPV-hsp70/lacZ) in orally inoculated larvae of Heliothis virescens was carried out. At dosages yielding similar final mortalities ( approximately 85 %), primary midgut infections by HzSNPV-hsp70/lacZ (indicated by lacZ expression) were observed 6 h earlier and in greater numbers than those generated by AcMNPV-hsp70/lacZ. Infection of secondary target cells in the tracheal epidermis, however, occurred at the same time and at the same rate for both NPVs. A 2 h delay was observed between the onset of primary and secondary AcMNPV-hsp70/lacZ infection, supporting the hypothesis that early tracheal infections were initiated by ODV nucleocapsids repackaged as budded virus. In contrast, an 8 h delay was observed with HzSNPV-hsp70/lacZ, suggesting that systemic infections were established only after virus replication in primary targets. Significant numbers of both MNPV- and SNPV-infected primary target cells were sloughed from the midgut beginning as early as 16 h post-infection. Midgut cell sloughing may be an important host-mediated selection pressure influencing the evolution of NPV morphology and gene regulation, shaping, in part, baculovirus infection strategies. PMID- 12560567 TI - Glycosylation inhibitors and neuraminidase enhance human immunodeficiency virus type 1 binding and neutralization by mannose-binding lectin. AB - Mannose-binding lectin (MBL), a C-type lectin component of the human innate immune system, binds to the gp120 envelope glycoprotein of human immunodeficiency virus type 1 (HIV-1). The objective of this study was to assess the effects of inhibitors of endoplasmic reticulum glucosidases and Golgi mannosidase as well as neuraminidase (NA) on the interaction between HIV and MBL. Production of HIV in the presence of the mannosidase I inhibitor deoxymannojirimycin (dMM) significantly enhanced binding of HIV to MBL and increased MBL neutralization of an M-tropic HIV primary isolate. In contrast, culturing HIV in the presence of alpha-glucosidase I and II inhibitors castanospermine and deoxynojirimycin only slightly affected virus binding and neutralization by MBL. Removal of sialic acid from HIV by NA also significantly enhanced virus binding and neutralization by MBL. Treatment of virus grown in the presence of dMM with endoglycosidase F1 substantially reduced binding to MBL, indicating that dMM increased MBL binding by increasing high-mannose carbohydrates on the virus. In contrast, endoglycosidase F1 did not decrease the MBL interaction with NA-treated virus, suggesting that NA exposed novel MBL binding sites. Treatment with dMM increased the immunocapture of HIV by monoclonal antibodies 2F5 and 2G12, indicating that altering the glycosylation of viral glycoproteins increases the accessibility or reactivity of some epitopes. This study shows that specific alterations of the N linked carbohydrates on HIV gp120/gp41 can enhance MBL-mediated neutralization of virus by strengthening the interaction of HIV-1 with MBL. PMID- 12560568 TI - Construction and immunogenicity in a prime-boost regimen of a Semliki Forest virus-vectored experimental HIV clade A vaccine. AB - A novel, experimental subunit human immunodeficiency virus (HIV) vaccine, SFV.HIVA, was constructed. This consists of Semliki Forest virus (SFV), which is a suitable vaccine vector for use in humans, and a passenger gene encoding HIVA, which is an immunogen derived from HIV-1 clade A that is being currently tested in clinical trials of combined DNA- and modified vaccinia virus Ankara (MVA) vectored vaccines in Oxford (UK) and Nairobi (Kenya). In the mouse, the SFV.HIVA vaccine was highly immunogenic for T cell-mediated immune responses and induced T cell memory that lasted for at least 6 months. SFV.HIVA was also compared to the vaccines currently used in the clinical trials and was shown to be as effective in T cell induction as pTHr.HIVA DNA but less immunogenic than MVA.HIVA. When tested in a prime-boost regimen, SFV.HIVA-induced responses could be boosted by MVA.HIVA. This work is a part of a long-term effort to build a panel of subunit vaccines expressing a common immunogen, which will allow both a direct comparison of various vaccine vectors and combined vaccination regimens in humans and provide more flexibility and/or a potential optimization of vaccinations for individuals based on their pre-existing anti-vector immunity. PMID- 12560569 TI - The proline-rich region of the ecotropic Moloney murine leukaemia virus envelope protein tolerates the insertion of the green fluorescent protein and allows the generation of replication-competent virus. AB - Sequences encoding the green fluorescent protein (GFP) were inserted into the envelope protein (Env) of ecotropic Moloney murine leukaemia virus, MoMLV. Insertion of these sequences into the proline-rich region (PRR) of Env resulted in a chimeric GFP-Env protein that allowed retrovirus vector transduction of murine cells with titres similar to wild-type Env. However, N-terminal extension with GFP did not result in a functional Env protein. GFP sequences were then inserted into the Env PRR of E-MO virus, a MoMLV that carries epidermal growth factor sequences at the N terminus of its Env protein. The resulting virus, GFP EMO1, replicates to the same titres as the parental virus. In a chronically infected cell culture, GFP-EMO1 was genetically stable. However, additional insertions of sequences that led to recombination or that may have been incompatible with virus replication were deleted and decreased virus titre. In summary, Env PRR can be used to tag individual virus particles with GFP, which leaves other regions available for modification in studies aimed at altering virus tropism. PMID- 12560570 TI - Walleye dermal sarcoma virus Orf C is targeted to the mitochondria. AB - Walleye dermal sarcomas are associated with the presence of a complex retrovirus, walleye dermal sarcoma virus (WDSV). These sarcomas develop and regress seasonally in naturally infected fish. In addition to gag, pol and env, WDSV contains three open reading frames (ORFs), designated orf a, orf b and orf c. orf c is located between the 5' long terminal repeat and gag. Developing tumours contain low levels of orf a and orf b transcripts, whereas regressing tumours contain high levels of genomic transcripts and virus particles. Orf C protein is encoded by the full-length, genomic transcript and can be detected in tumour extracts with anti-Orf C-specific antisera. To determine the subcellular location of WDSV Orf C, cultured cells were transfected with an expression vector encoding haemagglutinin-tagged Orf C and examined by immunofluorescence. Orf C was observed throughout the cytoplasm and accumulated in cytoplasmic organelles. Dual antibody staining for Orf C and mitochondrial cytochrome c demonstrated colocalization of Orf C with mitochondria and loss of the normal distribution of mitochondria in the cytoplasm. Cells transiently expressing Orf C exhibited apoptotic morphology and increased levels of surface phosphatidylserine and were unable to retain MitoTracker Orange, a dye that accumulates in active mitochondria. These results imply a functional role for WDSV Orf C in an alteration of mitochondrial function that results in apoptosis contributing to tumour regression. PMID- 12560571 TI - Translational regulation of rotavirus gene expression. AB - Rotavirus mRNAs are transcribed from 11 genomic dsRNA segments within a subviral particle. The mRNAs are extruded into the cytoplasm where they serve as mRNA for protein synthesis and as templates for packaging and replication into dsRNA. The molecular steps in the replication pathway that regulate the levels of viral gene expression are not well defined. We have investigated potential mechanisms of regulation of rotavirus gene expression by functional evaluation of two differentially expressed viral mRNAs. NSP1 (gene 5) and VP6 (gene 6) are expressed early in infection, and VP6 is expressed in excess over NSP1. We formulated the hypothesis that the amounts of NSP1 and VP6 were regulated by the translational efficiencies of the respective mRNAs. We measured the levels of gene 5 and gene 6 mRNA and showed that they were not significantly different, and protein analysis indicated no difference in stability of NSP1 compared with VP6. Polyribosome analysis showed that the majority of gene 6 mRNA was present on large polysomes. In contrast, sedimentation of more than half of the gene 5 mRNA was subpolysomal. The change in distribution of gene 5 mRNA in polyribosome gradients in response to treatment with low concentrations of cycloheximide suggested that gene 5 is a poor translation initiation template compared with gene 6 mRNA. These data define a regulatory mechanism for the difference in amounts of VP6 and NSP1 and provide evidence for post-transcriptional control of rotavirus gene expression mediated by the translational efficiency of individual viral mRNAs. PMID- 12560572 TI - Stable expression of antisense RNAs targeted to the 5' non-coding region confers heterotypic inhibition to foot-and-mouth disease virus infection. AB - The antiviral potential of transcripts targeted to the non-coding regions (NCRs) of foot-and-mouth disease virus (FMDV) RNA have been studied during transient and constitutive expression in susceptible BHK-21 cells. Transient expression of antisense transcripts corresponding to the 5' and 3'NCRs, alone or in combination, confers specific inhibition of homologous (serotype C) virus infection in BHK-21 cells. Constitutive expression of antisense 5'NCR transcripts (5'AS) exerted higher levels of inhibition to homologous and heterologous (serotypes O, A, Asia, SAT 1, SAT 2 and SAT 3) FMDV infection, as estimated by a 10-fold reduction in virus titre in the supernatants from infected clones and by a plaque reduction assay. These inhibitions were also observed, albeit to a lesser extent, in clones stably expressing antisense 3'NCR transcripts. The antiviral response was specific for FMDV, as the picornavirus encephalomyocarditis virus was not inhibited in any of the transformed cell lines. In all cases, a correlation was found between the level of transcript expression and the extent of virus inhibition. The potential to efficiently inhibit FMDV, including isolates representing the seven serotypes, by expressing interfering 5'AS transcripts opens the possibility of developing transgenic animals with a reduced susceptibility to FMDV. PMID- 12560573 TI - Interaction of glyceraldehyde-3-phosphate dehydrogenase with secondary and tertiary RNA structural elements of the hepatitis A virus 3' translated and non translated regions. AB - Proteins interacting with RNA structures at the 3' non-translated region (3'NTR) of picornaviruses are probably important during viral RNA replication. We have shown previously that a dominant cellular cytoplasmic protein of 38 kDa (p38) interacts with the 3'NTR and upstream regions of the hepatitis A virus (HAV) RNA (Kusov et al., J Virol 70, 1890-1897, 1996). Immunological and biochemical analyses of p38 have indicated that it is identical to GAPDH, which has previously been described as modulating translational regulation of the HAV RNA by interacting with the 5'NTR (Schultz et al., J Biol Chem 271, 14134-14142, 1996). Three separate binding regions for GAPDH in the 3'NTR and in the upstream 3D polymerase-coding region were identified. Structural analysis of these RNA regions by computer modelling and direct enzymatic cleavage suggested the presence of several AU-rich stem-loop structures having the potential for tertiary interactions. Binding of GAPDH to these structures was confirmed by RNA footprint analysis and resulted in the loss of double-stranded RNA regions. A different panel of RNA binding proteins (p28, p41 and p65) was detected in the ribosomal fractions of several cell lines (BSC-1, FRhK-4 and HeLa), whereas RNA binding of the GAPDH that was also present in these fractions was only marginal or absent. PMID- 12560574 TI - In vivo activity of Rhopalosiphum padi virus internal ribosome entry sites. AB - The RNA genome of Rhopalosiphum padi virus (RhPV), like other members of the Dicistroviridae, contains two open reading frames that are preceded by internal ribosome entry sites (IRESs). To compare the activities of the two RhPV IRESs in insect cells, a system was established for the in vivo transcription and translation of plasmid templates containing the IRESs. In this system, the two RhPV IRESs directed initiation of translation from bicistronic plasmids with equal efficiency. Competition was observed between the two IRESs when they were in cis in a bicistronic plasmid. A mutation that disrupted the 3'-proximal pseudoknot of the intergenic (IG) IRES reduced translation initiation in vivo. Similarly, mutations in the RhPV IG IRES disrupted its ability to bind 80S particles in vitro. The two IRESs preferentially labelled proteins of different masses in UV cross-linking experiments, illustrating the different translation initiation mechanisms employed by the two elements. PMID- 12560576 TI - The extent of homologous recombination in members of the genus Flavivirus. AB - The family Flaviviridae includes important human pathogens, such as dengue (DEN) virus, yellow fever (YF) virus and hepatitis C virus, many of which have emerged or re-emerged in recent years. Until recently, flavivirus evolution was thought to proceed in a clonal manner, with diversity generated mainly through the accumulation of mutational changes. However, this assumption has now been shown to be invalid, with homologous recombination demonstrated in all three genera of the FLAVIVIRIDAE: Since recombination has important implications for the study of virus evolution, a survey of recombination in the viruses of the genus Flavivirus was carried out. Using envelope gene sequence data and a combination of graphical and phylogenetic analyses, hitherto unreported recombination in Japanese encephalitis virus and St Louis encephalitis virus was detected, as well as further recombinants in DEN virus. However, no evidence for recombination was found in West Nile or YF viruses, or in the tick-borne flavivirus group. It is proposed that the difference between the mosquito- and tick-borne viruses can be accounted for by their differing modes of transmission, whilst the variation among the mosquito-borne flaviviruses reflects both the ecology of the particular host and vector species and also bias in the sampling process. PMID- 12560575 TI - Molecular and serological characterization of sporadic acute hepatitis E in a Japanese patient infected with a genotype III hepatitis E virus in 1993. AB - Serum samples collected periodically from a 40-year-old Japanese woman who had not travelled abroad and who had contracted sporadic acute hepatitis E in 1993 were semi-quantitatively tested by enzyme immunoassay for IgM, IgA and IgG antibodies to hepatitis E virus (HEV). Anti-HEV IgM and IgA antibody levels were the highest (1 : 2400 dilution and 1 : 3400 dilution, respectively) on day 9 after the onset of hepatitis and then decreased rapidly in a parallel manner. Anti-HEV IgG antibody levels were the highest (1 : 17000 dilution) on day 145 and then decreased gradually but remained at high titres (1 : 2200 dilution) even 8.7 years after the onset of hepatitis. An HEV isolate, HE-JA10, recovered from the patient's serum at admission was closely related to a genotype III strain isolated in the United States (US1), with 92.2% identity over the full-length genome, and was most closely related to the JMY-Haw isolate of Japanese origin (95.4% identity). PMID- 12560577 TI - Evolution of hepatitis C virus in blood donors and their respective recipients. AB - This paper describes the study of hepatitis C virus (HCV) evolution in the largest cohort of HCV-infected blood donors (BDs)/blood recipients (BRs) reported to date (25 pairs). A molecular analysis of partial sequences in the E1 (envelope) and NS5-B (polymerase) genes was performed. Phylogenetic reconstruction showed that the evolution of dominant strains was qualitatively and quantitatively different in BDs and BRs. The evolutionary rate was significantly higher in BRs, in which, in addition, most substitutions observed were antonymous. These findings corroborate the hypothesis that a large part of virus evolution - which was evaluated to be equivalent to approximately 20 years of chronic evolution - is acquired during the early phase of infection. These findings should be taken into account for the modelling of the long-term evolution of HCV and their possible contribution to improve our understanding of HCV natural history is discussed. PMID- 12560578 TI - Insertion of cellular sequence and RNA recombination in the structural protein coding region of cytopathogenic bovine viral diarrhoea virus. AB - The cytopathogenic bovine viral diarrhoea virus (cp BVDV) strain KS86-1cp was isolated from a calf persistently infected with the noncytopathogenic (ncp) strain KS86-1ncp after it was exposed to cp BVDV strain Nose and developed mucosal disease (MD). Molecular analysis revealed that an insertion of a cellular gene and a duplication of the viral RNA encoding the nucleocapsid protein C and part of N(pro) had occurred in the C coding region of the Nose and KS86-1cp genomes. The inserted cellular gene was closely related to the cINS sequence. Remarkably, the 5' upstream region from the insertion of KS86-1cp had high sequence identity to that of Nose, but differed from that of KS86-1ncp. In contrast, the region downstream from the insertion of KS86-1cp showed high identity to KS86-1ncp, but not to Nose. These data reveal that KS86-1cp is a chimeric virus generated by homologous RNA recombination in a calf with MD. PMID- 12560580 TI - The complete genomic sequence of hepatitis delta virus genotype IIb prevalent in Okinawa, Japan. AB - The Miyako Islands, located in the southernmost part of Japan, have been reported to be endemic for hepatitis delta virus (HDV). The majority of HDV patients in this area exhibit a relatively mild course of infection that evolves into a quiescent cirrhotic condition. The entire nucleotide sequence of the Miyako isolate (L215) of HDV obtained from a cirrhotic patient infected with HDV was determined. This isolate, L215, comprises 1682 nt and encodes 213 aa of the hepatitis delta antigen. Phylogenetic analysis showed that L215 is closely related to the Taiwanese genotype IIb HDV isolate. In addition, the predicted folding structure of the antigenomic RNA substrate was different from those of the published genotype II sequences. PMID- 12560579 TI - Upregulation of IL-10 gene expression in porcine peripheral blood mononuclear cells by porcine reproductive and respiratory syndrome virus. AB - Several lines of evidence suggest that porcine reproductive and respiratory syndrome virus (PRRSV) may have immunomodulatory effects on the host immune system. To determine the effect of PRRSV on cytokine production, a multiplex PCR was established. This allowed a semi-quantitative analysis of IFN-gamma, IL-2, IL 4, IL-10 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene expression levels from porcine peripheral blood mononuclear cells (PBMCs). These results showed that both live and inactivated PRRSV predominantly upregulated IL-10 gene expression in porcine PBMCs. In addition, when PBMCs from pigs immunized previously with classical swine fever virus (CSFV) vaccine were cultivated with the recall antigen, CSFV, in the presence of PRRSV, significant upregulation of IL-10 gene expression and reduction of IFN-gamma gene expression were observed. These findings indicated that the presence of PRRSV in the culture could affect recall antigen response. This study implies that the induction of IL-10 production may be one of the strategies used by PRRSV to modulate host immune responses. PMID- 12560581 TI - Phylogenetic relationships among members of the genus Phlebovirus (Bunyaviridae) based on partial M segment sequence analyses. AB - Viruses in the Phlebovirus genus of the family Bunyaviridae cause clinical syndromes ranging from a short, self-limiting febrile illness to fatal haemorrhagic fever. The genus currently consists of 68 antigenically distinct virus serotypes, most of which have not been genetically characterized. RT-PCR with four 'cocktail' primers was performed to amplify a region of the M segment of the genome of 24 phleboviruses included in the sandfly fever Naples, sandfly fever Sicilian and Punta Toro serocomplexes. Partial M segment sequences were successfully obtained and phylogenetic analysis was performed. The three resultant genotypic lineages were consistent with serological data. The sequence divergences were 27.6 % (nucleotide) and 25.7 % (amino acid) within the Sicilian serocomplex, 33.7 % (nucleotide) and 34.4 % (amino acid) within the Naples serocomplex and 35.6 % (nucleotide) and 37.5 % (amino acid) within the Punta Toro serocomplex. Overall, the diversities among viruses of Sicilian, Naples and Punta Toro serocomplexes were 48.2 % and 57.6 % at the nucleotide and amino acid levels, respectively. This high genetic divergence may explain the difficulties in designing a consensus primer pair for the amplification of all the phleboviruses using RT-PCR. It also suggests that infection with one genotype may not completely immunize against infection with all other genotypes in a given serocomplex. These findings have implications for potential vaccine development and may help explain clinical reports of multiple episodes of sandfly fever in the same individual. PMID- 12560582 TI - Effect of fusion protein cleavage site mutations on virulence of Newcastle disease virus: non-virulent cleavage site mutants revert to virulence after one passage in chicken brain. AB - Virulence of Newcastle disease virus (NDV) is mainly determined by the amino acid sequence of the fusion (F0) protein cleavage site. Full-length NDV cDNA clone pNDFL was used to generate infectious NDV with defined mutations in the F0 cleavage site (RRQRR downward arrow L, GRQGR downward arrow F, RRQGR downward arrow F, RGQRR downward arrow F and RKQKR downward arrow F). All the mutants were viable and the mutations were maintained after virus propagation in embryonated eggs. The mutants showed single-cell infections on chicken embryo fibroblasts, which suggested that they were non-virulent. However, virulence tests in 1-day old chickens resulted in an intracerebral pathogenicity index (ICPI) between 0 and 1.3. Moreover, virulent virus was isolated from chickens that had died in the virulence tests. Subsequent sequence analysis showed that the mutants RRQRR downward arrow L, RRQGR downward arrow F, RGQRR downward arrow F and RKQKR downward arrow F gave rise to the appearance of revertants containing the virulent cleavage site RRQ(K/R)R downward arrow F and an ICPI of 1.4 or higher. This indicated that reversion to virulence was caused by alteration of the amino acid sequence of the F0 cleavage site from a non-virulent to a virulent type. Furthermore, the ICPI of the revertants was higher than that of cDNA-derived strain NDFLtag, which has the same cleavage site, RRQRR downward arrow F (ICPI=1.3). NDFLtag(Pass), which was isolated from dead chickens after intracerebral inoculation of NDFLtag, also showed an increase in the ICPI from 1.3 to 1.5. This study proves that reversion to virulence occurs within non virulent NDV populations and that the virulence may increase after one passage in chicken brain. PMID- 12560583 TI - A molecular epidemiological study of Australian bat lyssavirus. AB - The genetic diversity of Australian bat lyssavirus (ABL) was investigated by comparing 24 ABL isolate glycoprotein (G) gene nucleotide sequences with those of 37 lyssaviruses representing Lyssavirus genotypes 1-6. Phylogenetic analyses indicated that ABL forms a monophyletic group separate from other lyssaviruses. This group differentiates into two clades: one associated with Pteropus (flying fox) species, the other with the insectivorous bat Saccolaimus flaviventris. Calculation of percentage nucleotide identities between isolates of the two clades revealed up to 18.7 % nucleotide sequence divergence between the two ABL variants. These observations suggest that ABL is a separate lyssavirus species with a similar epidemiology to chiropteran rabies virus (RV), where two distinct ABL variants co-exist in Australia in bat species with dissimilar ecology. Analyses of selection pressures in ABL G gene sequences provided some evidence of weak positive selection within the endodomain at amino acids 499 and 501, although in general the dominant evolutionary process observed was purifying selection. This intimates that, in nature, isolates of ABL, like those of RV, are subject to relatively strong selective constraints, suggesting a stability of host species, cell tropisms and ecological conditions. PMID- 12560584 TI - The catalytic subunit of protein kinase CK2 phosphorylates in vitro the movement protein of Tomato mosaic virus. AB - The movement protein (MP) of Tomato mosaic virus (ToMV) was reported previously by us to be phosphorylated in vitro by a cellular protein kinase(s) that exhibited several characteristics of casein kinase 2 (CK2). To characterize further this CK2-like cellular kinase, we have cloned cDNAs encoding the CK2 catalytic subunit from tobacco and compared the properties of the recombinant protein with those of the CK2-like cellular kinase. The recombinant CK2 catalytic subunit formed a complex with ToMV MP and phosphorylated it, similar to the CK2 like cellular kinase. Phosphoamino acid analyses of various mutant MPs altered near the C terminus revealed that the recombinant CK2 catalytic subunit phosphorylated serine-261, while the CK2-like cellular kinase phosphorylated both serine-261 and threonine-256. Both kinases were suggested to phosphorylate an additional serine residue(s) in regions other than the C-terminal peptide. The results are consistent with our previous prediction of involvement of CK2 in phosphorylation of ToMV MP. PMID- 12560585 TI - Treatment of small dense LDL. AB - The increased frequency of small, dense LDL is associated with the risk of coronary heart disease (CHD). Possible mechanisms include the increased susceptibility of small, dense LDL to oxidation and its high affinity for LDL receptor-independent cell surface binding sites. Although more than 30% of adult men in the USA have been reported to have small,dense LDL, only 5.4% of young Japanese men are affected. However, more than 76% of Japanese diabetics with coronary heart disease have small, dense LDL. Furthermore, almost half of all obese women (BMI > 35 kg/m(2)) have small, dense LDL. Our previous observation revealed that type 2 diabetics had smaller LDL even if they were apparently normolipidemic. In the normotriglycelidemic group there was also a close relationship between LDL size and plasma triglyceride. Diabetics with microalbuminuria had smaller LDL than those with normoalbuminuria, indicating the early nephrotoxicity of small, dense LDL. We also found that young men with high normal blood pressure have smaller LDL than those with optimal blood pressure. Furthermore, LDL size was decreased not only in preeclamptic women but also in normal pregnant women. Finally, weight reduction by obese women through strict diet control, the treatment of diabetics by acarbose or troglitazone, and the treatment of hyperlipidemia by new statins as well as fibrates were all successful in increasing LDL size associated with decreased plasma triglyceride. PMID- 12560586 TI - Gene therapy targeting monocyte chemoattractant protein-1 for vascular disease. AB - Monocyte chemoattractant protein-1 (MCP-1) has been shown to play an essential role in the pathogenesis of arteriosclerosis and other vascular diseases, such as restenosis after arterial injury, by recruiting monocytes into the arterial wall. We devised a new strategy for anti-MCP-1 gene therapy against arteriosclerosis by transfecting an amino-terminal deletion mutant (7ND), which lacks the amino terminal amino acids 2 to 8 of the human MCP-1 gene, into a remote organ (skeletal muscles). Intramuscular transduction with the mutant MCP-1 gene suppressed inflammatory and proliferative changes and arteriosclerosis formation induced by the chronic inhibition of nitric oxide synthesis in rats. 7ND gene transfection also inhibited the initiation, progression, and destabilization of atherosclerosis in Apolipoprotein E-knockout mice. Moreover, the strategy reduced restenosis after balloon injury in rabbits, rats, and monkeys, or neointimal formation after stent implantation in monkeys. This new strategy may be a useful and feasible gene therapy against atherosclerosis and restenosis after angioplasty. PMID- 12560587 TI - Insulin sensitizers and atherosclerosis. AB - It is surprising that only about ten years after the concept of insulin resistance in diabetes mellitus was established, the role of insulin resistance in the development of atherosclerosis has been discussed and clarified. Insulin resistance predisposes the development of glucose intolerance, hyperlipidemia, and hypertension; the cluster of these abnormalities is referred to as multiple risk factor syndrome and it increases the risk of atherosclerosis. A few insulin sensitizers have recently begun to be used in the therapy for diabetic patients. However, the inhibitory effects of these insulin sensitizers against atherosclerosis have not been studied in large-scale clinical trials because these drugs were approved for clinical treatment only several years ago. Accordingly, this review presents a summary of the previous studies on the anti atherogenic effects of insulin sensitizers by different strategies and provides information on why it is expected that insulin sensitizers will be used as anti atherogenic drugs. PMID- 12560588 TI - Insulin treatment prevents LDL from accelerated oxidation in patients with diabetes. AB - In a study population, we compared the level of malondialdehyde-modified LDL (MDA LDL) with the concentrations of lipid parameters in serum and found a strong correlation between MDA-LDL and apolipoprotein B (apo B) concentrations. Their interrelations had a turning point at an apo B concentration of 1,150 mg/l. In diabetic patients, the ratio of MDA-LDL/apo B increased at apo B concentrations above 1,150 mg/l. This ratio represents the extent of modification of apo B by MDA. In the control subjects, this ratio remained stable. When we divided the patients into medication groups (statins and insulin), we found that the 1,150 mg/l threshold disappeared. At apo B concentrations above 1,150 mg/l, the ratio of MDA-LDL/apo B in the statin group was as high as that in the non-drug group. In the insulin group, the means of MDA-LDL/apo B in all ranges of apo B levels decreased to an extent statistically indistinguishable from those of the control group. In conclusion, insulin therapy represses LDL oxidation even at apo B concentrations > 1,150 mg/l and should be noted for its anti-oxidation properties. PMID- 12560589 TI - Atheroembolic renal disease: clinical findings of 11 cases. AB - Atheromatous embolism is a systemic disease resulting from cholesterol crystal embolization in many organs, including the kidneys. To characterize atheroembolic renal disease (AERD), we retrospectively evaluated 11 patients with acute renal failure after vascular surgery, vascular radiology investigations, and anticoagulation at Miyazaki Medical College from 1994 to 2001. The diagnosis of cholesterol atheromatous embolism was confirmed by tissue examination or clinical grounds. The patients were all elderly men (average age of 66.8 years) with a history of hypertension (55%), diabetes mellitus (45%), hyperlipidemia (45%), and coronary artery disease (18%). Seven patients had livedo reticularis, and 4 had blood eosinophilia. Clinically, 7 patients were managed conservatively and 5 of them improved, whereas 4 patients required dialysis and developed chronic renal failure or died. The serum creatinine levels of the improved patients were significantly lower (1.28+/-0.3 mg/dl, p < 0.005) than the non-improved ones (7.70+/-3.6). The number of eosinophils was significantly higher in the improved patients (576+/-295 /ml, p < 0.05) than in the non-improved ones (208+/-206). However, no significant difference was observed in the levels of serum cholesterol and C-reactive protein among these patients. Since the population at risk for AERD is growing, we should recognize this disease as a cause of acute renal failure. PMID- 12560590 TI - Accelerated calcification represses the expression of elastic fiber components and lysyl oxidase in cultured bovine aortic smooth muscle cells. AB - Vascular calcification is a common feature of advanced atherosclerosis resulting in reduced elasticity of elastic arteries. However, the relationship between elastic fibers and vascular calcification at the molecular and cellular levels remains unknown. We investigated the expression of major elastic fiber components such as tropoelastin (TE) and fibrillin-1 (FBN1) and elastin-related enzyme, lysyl oxidase (LO), in a calcification model using beta-glycerophosphate (beta GP) in cultured bovine aortic smooth muscle cells (BASMCs). Ten mM of beta-GP stimulated calcium deposition in a time-dependent manner. As determined by Western blot analysis, 10 mM of beta-GP time-dependently decreased TE and FBN1 protein levels. TE, FBN1, and LO mRNA levels, assessed by reverse transcription polymerase chain reaction, were also decreased by exposure to 10 mM beta-GP. Furthermore, we investigated whether the processes of calcification in BASMCs directly control these regulations. In experiments using levamisole, an alkaline phosphatase inhibitor, and DMDP, a bisphosphonate, both inhibitors inhibited down regulation during beta-GP-induced calcification, suggesting that the down regulation of TE, FBN1, and LO directly relates to calcium deposition. In cases of vascular calcification, the decreased expression of TE, FBN1, and LO may be partially responsible for decreased vascular elasticity and also for the decreased formation of new elastic fibers. PMID- 12560591 TI - Influence of the extent of westernization of lifestyle on the progression of preclinical atherosclerosis in Japanese subjects. AB - To clarify the influence of a westernized lifestyle on the risk factors for atherosclerosis and preclinical atherosclerosis in Japanese subjects, we surveyed a Japanese population and Japanese immigrants in the United States. Based on the extent of westernization of their lifestyle, the subjects were classified as Japanese (J), first generation Japanese-Americans (JA-I), and second or later generation Japanese-Americans (JA-II). The consumption of animal fat and simple carbohydrates increased in the order of J, JA-I, and JA-II, while the subjects with strenuous physical activity decreased in the same order. The waist-hip ratio, fasting insulin level, serum cholesterol and triglyceride levels, and prevalence of hypertension increased in the same order as the dietary changes. The carotid intima-media wall thickness and the plaque size, which are indices of preclinical atherosclerosis, also increased in the order of J, JA-I, and JA-II. These data indicate that a westernized lifestyle aggravates the risk factors for atherosclerosis and influences the progression of preclinical atherosclerosis, in correspondence with the extent of westernization. PMID- 12560592 TI - Effect of diabetes on uremic dyslipidemia. AB - Elevated intermediate-density lipoprotein (IDL), a remnant lipoprotein, is an independent risk factor for atherosclerosis in patients with end-stage renal disease (ESRD). Since the presence of diabetes mellitus further increases the risk of cardiovascular mortality in ESRD, we examined the effect of diabetes on IDL among ESRD patients. The subjects were 330 healthy control subjects and 287 patients with end-stage renal disease including 80 patients with type 2 diabetes. As compared with the healthy subjects, the nondiabetic ESRD patients had increased plasma triglyceride and IDL cholesterol. Diabetic patients with ESRD showed a further increase in plasma triglyceride and IDL cholesterol compared with the nondiabetic group. However, the difference in IDL levels between the ESRD groups was no longer significant when subjects were stratified by plasma triglyceride. Plasma triglyceride was correlated with IDL cholesterol. Increased hemoglobin A(1c) was significantly associated with IDL cholesterol in a multiple regression model including age, gender, and the presence of ESRD. Such an association was no longer significant in another model including plasma triglyceride as an additional covariate. Further analysis indicated the positive effects of diabetes and hyperglycemia on plasma triglyceride. These results indicate that increased IDL in ESRD is further deteriorated in the presence of diabetes, and that the adverse effect is accounted for at least partly by hypertriglyceridemia associated with chronic hyperglycemia. PMID- 12560593 TI - Familial combined hyperlipidemia (FCHL) in children: the significance of early development of hyperapoB lipoproteinemia, obesity and aging. AB - It is not easy to make diagnose FCHL in children, since a clear expression of lipoprotein abnormality is unlikely and standard criteria have not yet been established. We investigated eight cases of childhood FCHL and their families with respect to familial history, anthropometric parameters and serum lipoprotein levels, to explore the characteristics of childhood FCHL. To diagnose childhood FCHL it is necessary to clarify both the family history and lipid profiles of the parents. In this study, two prominent features were suggested; that serum TG level is affected by both obesity and age, and also in particular, that a significantly elevated level of serum apoB is a predominant feature of FCHL in childhood. It was found that hyperapoB may be revealed antecedently without other lipid abnormalities at an early age. Regardless of other lipoprotein abnormality, it was suggested that hyperapoB might be added to the early diagnostic criteria for FCHL. PMID- 12560594 TI - Dynamical aspects of in vitro conditioning in Hermissenda type B photoreceptor. AB - Dynamics of changes in physiology and morphology were studied in Hermissenda photoreceptors after in vitro conditioning with paired light and vibration. An increase in input resistance of the type B photoreceptor was observed following 5 paired presentations of light and vibration. It peaked at 10 min after in vitro conditioning, then decreased to a level twice the pre-conditioning level for more than 60 min. Contraction of the terminal branches along centro-lateral direction was initiated 5 min after conditioning and reached its final state at 10 min after conditioning. The pairing specific contraction of the axon terminal was not observed in ASW containing anisomycin. The dynamics in physiology and morphology were completely parallel 30 min after conditioning. These findings suggested that in vitro conditioning induced contraction was dependent on protein synthesis dependent process initiated within 5 min after training trials and that the change of cell morphology is a form of short-term synaptic plasticity that involves changes in macromolecular synthesis. Present findings that functional remodeling at the terminal branch of the type B photoreceptor occurred within 10 min after conditioning was the fastest modification process reported so far. PMID- 12560596 TI - Sensitization and habituation of the swimming behavior in ascidian larvae to light. AB - Ascidian larvae of Ciona intestinalis change their photic behavior during the course of development. Newly hatched larvae show no response to a light stimulus at any intensity. At 4 hr after hatching, larvae were induced to start to swimming upon the cessation of illumination, and to stop swimming upon the onset of illumination. At a weaker light intensity (5.0 x 10(-3) J/m (2).s), the larvae showed similar responses to either a single stimulus or repeated stimuli of onset and cessation of light until 10 hr after hatching. At a stronger light intensity (3.2 x 10(-1) J/m(2).s), when the stimulus was repeated, they showed sensitization and habituation of the swimming response. At 3 hr after hatching the larvae failed to show any response to an initial stimulus at any intensity of light, but after several repeated stimuli (sensitization) they showed a swimming response at light intensities above 4.0 x 10(-2) J/m (2).s. At 5 hr and with intensity above 1.0 x 10 (-2) J/m(2).s, the larvae showed photoresponses to the first stimulus, but after several repetitions the larvae failed to stop swimming upon the onset of light (habituation). A repeated series of stimuli at stronger intensities of light caused greater habituation; this habituation was retained for about 1 min. Since the larval central nervous system in Ciona is comprised of only about 100 neurons, learning behavior in ascidian larvae should provide insights for a minimal mechanism of memory in vertebrates. PMID- 12560595 TI - Presynaptic K+ channel modulation is a crucial ionic basis of neuronal damage induced by ischemia in rat hippocampal CA1 pyramidal neurons. AB - The ischemia-induced synaptic potentiation (ISP) during and/or after brain ischemia has been suggested to be one of the crucial factors responsible for irreversible neuronal damage of hippocampal CA1 pyramidal neurons. However, the presynaptic modulation mechanism that leads to neuronal damage during and/or after ischemia was still unknown. By combining electrophysiological methods and infra-red differential interference contrast (IR-DIC) imaging procedures, we showed for the first time that ISP is the result of extraordinary presynaptic depolarization in association with the suppression of 4-aminopyridine (4-AP) sensitive K(+) channels at the presynaptic sites. Furthermore, we also showed that the 4-AP sensitive presynaptic K(+) channels played a crucial role in inducing neuronal damage at a very acute phase of ischemia-induced neuronal damage and would be a therapeutic target against the neuronal damage after brain ischemia. PMID- 12560597 TI - The cooperativity of human fetal and adult hemoglobins is optimized: a consideration based on the effectiveness of the Bohr shift. AB - The physiological significance of the cooperativity of human hemoglobin (Hb) is considered from the viewpoint of the effectiveness of the Bohr shift at the sites of O(2) release and uptake across the placental membrane. The effects of the Bohr shift was examined by changing the O(2) saturation of Hb (S(pO2)) per unit change in P(50), -dS(PO2)/d P(50), where P(50) is partial pressure of O(2) at half saturation. The Bohr shift at the sites of O(2) uptake and release was found to be highly effective in both fetal and maternal bloods at physiological degree of cooperativity (Hill's coefficient, n=2.65). From the results obtained in this paper, it is concluded that the positions of OECs of fetal and maternal Hbs are regulated to receive a maximal benefit from the Bohr shift, and that a relatively low n value of human tetrameric Hb is adequate for the O(2) and CO(2) exchange across the placental membrane. PMID- 12560598 TI - Ambient salinity modulates the expression of sodium pumps in branchial mitochondria-rich cells of Mozambique tilapia, Oreochromis mossambicus. AB - Na,K-ATPase (sodium pumps) provide the primitive driving force for ion transport in branchial epithelial cells. Immunoblots of epithelial homogenates of both seawater (SW)- and freshwater (FW)-adapted tilapia gills as well as rat brain homogenate, a positive control, revealed one major band with a molecular weight of about 100 kDa. SW-adapted tilapia gills possessed larger (about 2-fold) amounts of sodium pumps compared with FW-adapted tilapia gills. (3)H-ouabain binding representing functional binding sites of Na,K-ATPase was also higher (about 3.5-fold) in gills of SW-adapted tilapia compared to that of FW-adapted fish. Moreover, specific activities of SW fish were higher (about 2-fold) than those of FW fish. Double labeling of Na,K-ATPase and Con-A, a fluorescent marker of MR cells, in tilapia gills followed by analysis with confocal microscopy showed that sodium pumps were localized mainly in MR cells, including the SW type and different FW types. Although more-active expression of Na,K-ATPase was demonstrated in gills of SW-adapted tilapia, no significant differences in densities of apical openings of MR cells were found between SW- and FW-adapted fish. These results indicate that, during salinity challenge, tilapia develop more "functional" Na,K-ATPase in SW-type MR cells to meet physiological demands. PMID- 12560599 TI - Molecular characterization of a cDNA encoding putative vitellogenin from the Pacific oyster Crassostrea gigas. AB - To elucidate the molecular mechanisms involved in oogenesis, we applied a differential display method to identify genes whose expression was detected only in ovaries containing oocytes. One of the cDNA fragments isolated by mRNA differential display was similar in structure to vitellogenin. Using this fragment, a full-length cDNA encoding putative vitellogenin in the Pacific oyster Crassostrea gigas was cloned by RACE (rapid amplification of cDNA ends), and its amino acid sequence was deduced. The open reading frame predicted 1583 amino acid residues. The deduced primary structure of putative vitellogenin in C. gigas was shown to be similar to vitellogenins of various other mollusk, fish, crustacean and nematode species, especially in the N-terminal region. Reverse transcription mediated PCR revealed that mRNA encoding putative vitellogenin was expressed only in the ovary. In situ hybridization analysis revealed that putative vitellogenin mRNA was expressed strongly in the follicle cells in the ovary. It is concluded that the follicle cells are the site of putative vitellogenin synthesis. PMID- 12560600 TI - No effects of estrogen receptor overexpression on gonadal sex differentiation and reversal in medaka fish. AB - In order to elucidate a possible role of estrogen receptor in the gonadal sex differentiation and the sex reversal with sex steroids, we examined for the formation of testis or ovary in transgenic medaka fish overexpressing the medaka estrogen receptor under the constitutive medaka beta-actin promoter. The transgenic fish underwent the genetically determined gonadal differentiation and showed the same sex-reversal rates as those of wild-type non-transgenic fish after treatments with estrogen and androgen. These results present invaluable data to reconsider the role of estrogen receptor in the gonadal sex determination. PMID- 12560601 TI - Life history of Milnesium tardigradum Doyere (tardigrada) under a rearing environment. AB - A strain of carnivorous tardigrade, Milnesium tardigradum, was reared in water on agar plates at 25 degrees C. The monogonont rotifer Lecane inermis was presented as a food source. This rearing system permitted detailed observation of tardigrade behaviour. Daily measurements of body length allowed the growth rate and moulting cycle of this species to be determined. The life history of M. tardigradum raised under these conditions included up to seven periods of moult. The first and second moults occurred at intervals of 4-5 days, and individuals reached reproductive maturity at the 3rd-instar stage; the first period of egg laying accompanied the third moult. The most rapidly developing animal in the study population laid eggs 12 days after hatching. The egg-laying intervals or moulting intervals of adult animals were around 6-10 days. The mean clutch size was 6.9 eggs. All tardigrades in this laboratory population were female and reproduced by parthenogenesis. The duration of the embryonic stage ranged from 5 16 days. The most long-lived female survived for 58 days after hatching, and laid a total of 41 eggs in 5 separate clutches. The entire life cycle of tardigrades reared under these conditions was recorded and photographed. A brief description of the embryonic development of M. tardigradum was also reported. PMID- 12560603 TI - In vitro induction of luteinizing hormone synthesis by estrogen in organ-cultured pituitary glands of the Japanese eel, Anguilla japonica. AB - An organ culture method for pituitary glands isolated from immature Japanese eels (Anguilla japonica) was developed. This method could conserve the histological features of the pituitary glands for at least 21 days. The ability to synthesize gonadotropic hormone (GTH) in cultured eel pituitary glands was examined by detecting luteinizing hormone (LH) beta protein immunohistochemically. In a basal medium (Leibovitz L-15), LH beta-immunoreactive cells were very scarce, but after addition of estradiol-17beta (E2) a large number of immunoreactive cells appeared, particularly in the proximal pars distalis. The stimulatory effects of E2 on LH beta synthesis were dose (1-100 ng/ml)- and time (1.5-7 days)-dependent. Thus, in contrast with previous reports of the lack of a direct effect of E2 on GTH synthesis in primary cultured eel pituitary cells, the present results clearly indicate that E2 can stimulate GTH synthesis in immature eel pituitary glands. This organ culture method is useful to examine the actions of steroids and also other endocrine factors on the eel pituitary gland. PMID- 12560602 TI - Expression pattern and transcriptional control of SoxB1 in embryos of the ascidian Halocynthia roretzi. AB - The Sox family is a large group of transcription factors that are characterized by the presence of a DNA-binding HMG domain. We isolated HrSoxB1, an ascidian homolog of the Sox gene that belongs to the B1 subclass of the Sox family, from Halocynthia roretzi. Expression was initiated as early as the 8-cell stage. During cleavage stages, HrSoxB1 was expressed in three quarters of embryonic blastomeres but not in posterior-vegetal (B-line) blastomeres. Misexpression of mRNAs of HrPEM but not of macho-1, whose maternal mRNAs are localized to the posterior-vegetal cytoplasm of eggs and early embryos, repressed the anterior vegetal expression of HrSoxB1. This result suggests that the zygotic expression of HrSoxB1 is controlled by the localized maternal mRNA. When HrSoxB1 was overexpressed in early embryos, ectopic expression of HrBra, a gene for a transcription factor expressed in notochord blastomeres, occurred in the most posterior blastomeres (B7.5), although these blastomeres did not eventually differentiate into notochord but developed into muscle, as they do in normal embryogenesis. In later embryogenesis, HrSoxB1 was specifically expressed in neural plate cells. However, overexpression of HrSoxB1 did not affect the expression of a neural plate marker gene, HrETR-1. PMID- 12560604 TI - Molecular cloning and expression analysis of cDNAs encoding androgenic gland hormone precursors from two porcellionidae species, Porcellio scaber and P. dilatatus. AB - Male sexual characteristics in Crustacea are induced by androgenic gland hormone (AGH), which is produced by the male-specific androgenic gland. Recently, AGH in the terrestrial isopod Armadillidium vulgare was characterized and its cDNA cloned, the first example in which the structure of AGH was elucidated. We report here the molecular cloning of cDNAs encoding AGH precursors from two additional terrestrial isopods, Porcellio scaber and P. dilatatus. cDNA fragments encoding Porcellio scaber AGH (Pos-AGH) and P. dilatatus AGH (Pod-AGH) were amplified by RT-PCR using degenerate oligonucleotide primers designed based on the amino acid sequence of A. vulgare AGH (Arv-AGH). Subsequently, full length cDNAs were obtained by 5'- and 3'-RACE. Both AGH precursors consisted of a signal peptide, B chain, C peptide and A chain, and exhibited the same organization as that of Arv AGH. The amino acid sequences of the A and B chains, which comprise mature AGH peptide, were highly conserved among the three species, while that of the C peptide showed only low sequence similarity. In Northern blot analysis, each cDNA fragment used as a probe specifically hybridized with a single band (0.75 kb) in mRNA extracted from whole male reproductive organs. In analysis of the tissue specific gene expression of these two AGHs by RT-PCR, it was revealed that both AGH transcripts were detected only in cDNA synthesized using total RNA from the testis carrying the androgenic glands, but not in that from testis only, seminal vesicle, vas deferens, or hepatopancreas. PMID- 12560605 TI - Gene expression and the physiological role of transforming growth factor-alpha in the mouse pituitary. AB - Transforming growth factor-alpha (TGF-alpha), a member of the epidermal growth factor (EGF) family, is produced within the mouse anterior pituitaries. However, the cell types of TGF-alpha-expressing cells and the physiological roles of TGF alpha within mouse pituitary glands remain unclear. The aim of the present study was to localize TGF-alpha mRNA-expressing cells, and to clarify the involvement of TGF-alpha in estrogen-induced DNA replication in mouse anterior pituitary cells. Northern blot analysis demonstrated TGF-alpha mRNA expression in adult male and female mouse anterior pituitaries. In situ hybridization analysis of the pituitaries in these mice showed that TGF-alpha mRNA-expressing cells in the anterior pituitary are round, oval, and medium-sized. TGF-alpha mRNA was colocalized in most of the growth hormone (GH) mRNA-expressing cells, while only some of the prolactin (PRL) mRNA-expressing cells. DNA replication in the anterior pituitary cells was detected by monitoring the cellular uptake of a thymidine analogue, bromodeoxyuridine (BrdU) in a primary serum-free culture system. Estradiol-17beta (E2) and TGF-alpha treatment increased the number of BrdU-labelled mammotrophs, indicating that E2 and TGF-alpha treatment stimulates the DNA replication in mammotrophs. Immunoneutralization of TGF-alpha with anti TGF-alpha-antibodies nullified the E2-induced increase in DNA replication. RT-PCR analysis of TGF-alpha mRNA expression in ovariectomized female mice revealed that E2 increases TGF-alpha mRNA levels. These results indicate that the TGF-alpha produced primarily in the somatotrophs mediates the stimulatory effects of estrogen on the DNA replication of pituitary cells in a paracrine or autocrine manner. PMID- 12560606 TI - Phylogenetical positions of Sorex sp. (Insectivora, Mammalia) from Cheju Island and S. caecutiens from the Korean Peninsula, inferred from mitochondrial cytochrome b gene sequences. AB - Phylogenetical positions of Sorex specimens unassigned to species from Cheju Island, Korea, and S. caecutiens from southern Korean Peninsula were investigated based on full nucleotide sequences (1,140 bp) of the mitochondrial cytochrome b gene, comparing specimens of the S. caecutiens/shinto group from locations throughout its range. In the phylogenetic tree obtained, S. caecutiens were separated into two main groups: Hokkaido and Continent-Sakhalin-Cheju clusters. Shrews from Cheju and Korean Peninsula were included in the latter cluster. Thus, we suggest that the shrew on Cheju Island should be ranked as S. caecutiens, although taxonomic description of the shrew has to be conducted elsewhere. The Cheju shrews formed a single sub-cluster while the peninsular shrews were not included in a single sub-cluster. The clustering of individuals in Continent Sakhalin-Cheju cluster did not always reflect the geographical proximity of their capture locations. We interpret these findings as indicating ancestral isolation of a Hokkaido population and recent rapid range expansion of the modern population in Eurasian Continent-Sakhalin-Cheju. PMID- 12560607 TI - Genetic variation and differentiation in the Japanese five-lined skink, Eumeces latiscutatus (Reptilia: Squamata). AB - The genetic variation in Eumeces latiscutatus from the main islands of Japan and the northern Ryukyus was investigated and compared with that of its close relatives (E. okadae and E. marginatus) using allozyme data. All three samples from the northern Tokara Island Group, currently identified as E. latiscutatus, were shown to belong to E. marginatus and not to E. latiscutatus. The non monophyly of the northern Tokara samples and the great genetic differences within these samples may have resulted from colonization from more than one source population by northeastward overseas dispersal. The samples from the Izu Peninsula were genetically much closer to E. okadae than the other conspecific ones. This indicates that the samples from the Izu Peninsula and the other samples of E. latiscutatus should be treated as distinct species, and that E. latiscutatus from the Izu Peninsula and E. okadae from the Izu Island Group may be treated as conspecific. Samples from western Japan were genetically well differentiated from those of eastern Japan. Within the western group, the samples from the Osumi Island Group was genetically distinct from those from the other regions, by possessing unique alleles. Our phenograms also reveal a distant affinity between samples from the Danjo Island Group and the main islands of Japan. This may be the result of long geographic isolation of the Osumi and Danjo Island Groups from Kyushu. By contrast, samples from Sapporo and Aomori were poorly differentiated genetically in spite of the long separation of these two localities by the Tsugaru Strait. This suggests that overseas dispersal of E. latiscutatus occurred across this strait after its formation. PMID- 12560608 TI - [The surgeon and deontology]. AB - The aim of study is to recall surgeons deontological principles and errors. The article demonstrates some specific deontological errors, performed by surgeon on patients and his colleagues; points out painful sequela of these errors as well. CONCLUSION. The surgeon should take in account deontological principles rigorously in routine daily practice. PMID- 12560609 TI - [History of Lithuanian heart surgery]. AB - The aim of the article is to review Lithuanian cardiac surgery development, as well as authors and timing of the first operations performed. Historical review involves two periods: initial (1900-1964) and contemporal (1964-2002). Benevolent rivalry between Vilnius and Kaunas heart surgeons is given to show their contribution into Lithuanian surgery. Chronological development and amelioration of the main heart surgery fields are described: heart injuries, ischemic heart disease, congenital and acquired defects, heart rhythm disturbances, heart transplantation. PMID- 12560610 TI - [Treatment strategy of unstable angina and non-ST-segment elevation myocardial infarction]. AB - Treatment strategy of unstable angina and non-ST-segment elevation myocardial infarction according to American College of Cardiology / American Heart Association practice guidelines (2002) is described in this article. PMID- 12560611 TI - [Surgical treatment of pulmonary emphysema]. AB - Lung volume reduction has become an accepted option for surgical treatment of pulmonary emphysema. Lately this method was introduced in Lithuania.The first 14 of 19 patients, who underwent lung volume reduction surgery, where admitted due to spontaneous pneumothorax, that is a common complication of pulmonary emphysema. Lung volume reduction using wedge resection or plication was performed in 13 and 6 patients respectively. Rethoracotomy was required for postoperative bleeding (2 cases) and prolonged air leakage (1 case). Two patients died. Pulmonary insufficiency decreased significantly in 2/3 of operated on patients. There were no recurrent spontaneous pneumothorax. PMID- 12560612 TI - [Tracheobronchial surgery]. AB - During the flowering of cardiothoracic surgery over the past 50 years, surgery of the major airway failed to develop correspondingly. The relative rarity of such cases accounts in past for this laggardness. Surgical diseases of the trachea, whether inflammatory or neoplastic, largely are presented as an obstructive problems of the airway. Surgical management of these lesions is based on simple concept of resection of the involved area of the trachea, when the larynx has not been lost because of affection by the primary disease. Primary end-to-end reconstruction of the trachea has been generally recognized as the ideal method of repair following resection. However, based on Belsey's experience it was widely believed, that only 2 cm at most could be removed and the trachea reconstruction by end-to-end suture in any dependable fashion. During the period of 30 years tracheobronchial surgical reconstructions have been accomplished in 187 patients. CONCLUSION: Management of the patients with stenosis of the trachea and main bronchi must be started promptly. Emergency treatment for the patients with severe tracheal stenosis is rigid bronchoscopy under general anesthesia. Circular tracheal resection is the best method of radical treatment for patients with benign and malignant tracheal narrowing. Sleeve resection is the ideal form of excisional therapy for benign endobronchial tumors, neoplasms of low-grade malignant potential, and bronchostenosis. For patients with tracheobronchial stenosis who are no candidates for surgical reconstruction, tracheobronchial stenting is the management of choice. PMID- 12560613 TI - [Bronchoplastic operations for lung cancer]. AB - Objective of our work was to evaluate efficacy of bronchoplastic operations for lung cancer and time to progression in combined treatment. From 1997 till 2001, 57pts were operated for early I-IIB stages of lung cancer. Operations were: tracheal resections in 3pts (5.2%), window right pneumonectomies in 5pts (8.7%), window left pneumonectomies in 2pts (3.5%), window right upper lobe in 22pts (38.5%), bifurcation resections 2pts (3.5%), sleeve right upper lobe resections 7pts (12.2%), sleeve left upper lobe resections in 11pts (19.2%). We had complications: in 7pts (12.2%) suture failure, 26pts (45.6%) obstructive pneumonia, 3pts (5.2%) kinking of anastomosis, 2pts (3.7%) bronchial bleeding, 6pts (10.5%) covered bronchial fistulas, 5pts (8.7%) died after operations. RESULTS: 32pts (56%) underwent radiation after surgery, 13pts (22.8%) radiation and chemotherapy. Three-year survival was in 82.4% (47pts), in 10pts (17.4%) disease progressed. CONCLUSIONS: 1. Bronchoplastic operations are sufficient for early lung cancer treatment. 2. Three-year was in survival 82.7% of pts. Seventeen percent of patients failed after combined treatment. PMID- 12560614 TI - [Pulmonary sequestration]. AB - Pulmonary sequestration is rare and serious dysplasia, which has clinical signs of pneumonia and respiratory distress syndrome and radiological signs of polymorphic shadows. Pulmonary sequestration masses have autonomic blood supply. For confirmation of the diagnosis we use ultrasound examination, CT scan, MRI, angiopulmography and aortography. AIM OF THE STUDY: To present pathogenesis, diagnostic features and surgical treatment tactics of rare and serious pulmonary dysplasia. MATERIAL AND METHODS: During the period of 1979-2002, we have examined and treated 9 patients because of pulmonary sequestration. We analyzed clinical signs, results of the examination before operation, indications for operation and results of surgical treatment. RESULTS: Seven patients after the operation recovered, 1 patient died due to infection and cardiovascular complications. One patient is supposed to be operated on in the near future. CONCLUSIONS: 1. Pulmonary sequestration is rare and serious bronchial - pulmonary dysplasia, which has clinical signs of pneumonia and respiratory distress syndrome and radiological signs of polymorphic shadows. 2. Pulmonary sequestrations are classified as intralobar, extralobar and extrapulmonary sequestration. 3. Pulmonary sequestration is often associated with cardiovascular dysplasias and diaphragmatic malformations. 4. Methods of examination: X-ray examination, CT scan, MRI, ultrasound examination, aortography. 5. Complications of pulmonary sequestration: pneumonia, pleuritis, necrosis of the sequestration, chylothorax, respiratory distress syndrome, hemothorax, cystic transformation, and pneumothorax. 6. Treatment - surgery or embolisation of abnormal supply vessels. PMID- 12560615 TI - [The surgical treatment of lung tuberculosis]. AB - The article informs about the treatment results of 1036 patients operated on because of different forms of pulmonary tuberculosis. According to spreading of tuberculosis and developed complications in spite of the medicamentical treatment the patients were divided into two clinical groups. The first group included 387 patients with direct and relative indications for surgery. The second group included 649 patients with complications of pulmonary tuberculosis, and 167 new diagnosed cases were documented among them (with spontaneous pneumothorax - 180, spontaneous pyopneumothorax - 117, tuberculous pleural empyema - 225, pulmonary hemorrhage - 101, with pulmonary reoperations - 26). The results of radical and paliative operations were as follows: 991 (95.6%) patients recovered, 45 patients (4.4%) died. CONCLUSION: When the therapeutical treatment of pulmonary tuberculosis is ineffective, especially in drug-resistant cases, the surgical treatment is indicated. PMID- 12560616 TI - [Initial experience of thoracic surgery in Klaipeda regional Hospital]. AB - OBJECTIVES: Study and profound analysis of primary clinical results of the recently established department of Thoracic surgery. Klaipeda Regional Hospital currently functions as a multiprofile facility and covers Western area of Lithuania. More than 274 patients were cured here within the past 8 months. Majority of patients, i.e. 205 (74.82%) were operated; 26 (12.68%) of them underwent surgery urgently and 179 (87.32%) - on planned basis. A large scale of different interventions was performed: 156 (76.1%) - complex operations of lungs, pleura, esophagus, mediastinum and others. All the rest - 49 (23.9%) - operations with minimally invasive procedures, i.e. thoracoscopy, mediastinoscopy, tracheostomy, esophagus stenting etc. CONCLUSIONS: Primary experience of thoracic surgery in Klaipeda regional hospital has good basis for the future development of this activity. PMID- 12560617 TI - [Stair climbing test in prediction of postoperative complications after lung cancer surgery]. AB - Preoperative physical state of a patient is very important for adaptation of the patient after lung resections. Purpose of this work is to evaluate an information factor of a stair-climbing test while predicting of postoperative complications after lung cancer surgery. Fifty two patients were examined, who passed lung surgery of different volume. The patients are distributed to two groups: I(st) group included the patients able to climb 1-44 footsteps (n=22/42.3%) and the II(nd) group included the patients able to climb more than 44 footsteps at a moderate speed without stopping for rest (n=30/57.7%). One flight of stairs made up to 22 footsteps with 15 cm of height each. Postoperative myocardial ischemia, disorders of heart rhythm, pneumonias, atelectasis, prolonged artificial ventilation of lungs, sanative bronchoscopy, duration of treatment, and cases of death were registered. It was established that postoperative cardiac and lung complications occurred in 17 patients (32.7%), two patients died (3.8%). Rate of complications between the patients of the I(st) and II(nd) group was 82.4 ir 17.6 percent. Postoperative course was normal for those patients (n=11) who were able to climb five or more flights of stairs. It was noticed that duration of postoperative period has an inverse proportion to a number of the climbed up footsteps. The stair-climbing test is a simple, safe, cheap and informative enough for prediction of postoperative cardiopulmonary complications after lung cancer surgery. PMID- 12560618 TI - [Characteristics of postoperative course in cancer patients]. AB - Objective of our work was to analyze postoperative care peculiarities in elderly cancer patients over 80 years. A retrospective analysis is made in connection with postoperative course in 152 cancer patients. The I(st) group included patients over the age 80 years (8.7 years in average), and the II(nd) group included patients of 60 years. Distribution of the patients into groups was equivalent as per volume and duration of operations. We have established that postoperative complications in patients of the I(st) and II(nd) group were noticed as 46.0 and 12.5 percent accordingly, that included 22.5 and 2.8 percent of lung complications, 23.7 and 4.2 percent of cardiovascular complications. Episodes of disorientation (delirium) were noticed in every fifth patient over the age of 80 years. Postoperative death-rate made up to 13.7 percent in the I(st) group, and 2.8 per cent in the II(nd) one. Results of the performed work showed that surgical treatment of cancer patients over the age of 80 years is related to a higher risk. Therefore, indications for operation and its volume has to be defined strictly taking into consideration preoperative functional status of the patients. CONCLUSIONS: 1. Postoperative complications three times and mortality five times are higher in cancer patients over 80 years to compare to 60 year-old patients. 2. Preoperative live functions status influence more to postoperative mortality than age as independent risk factor. PMID- 12560619 TI - [Possibility of computed tomography in evaluation of locoregional invasion of non small cell lung cancer]. AB - Clinical prognosis and treatment schedules of non-small cell lung cancer are dependent on tumor stage. The objective of the study was to assess the diagnostic value of computed tomography scans in preoperative staging of non-small cell lung cancer. METHODS: One hundred thirty seven case reports were analyzed retrospectively and 72 cases were found eligible for the analysis. RESULTS: The sensitivity, specificity, positive and negative prognostic values of computed tomography in assessing metastases in mediastinal lymphnodes were 0.79; 0.45; 0.34; 0.85 respectively. The overall accuracy was 0.51. When evaluating the direct invasion of the tumor to chest wall, pericardium and mediastinum the values were as follows: 0.56; 0.87; 0.59; 0.86; 0.76. The conclusion is made that the accuracy of computed tomography in evaluating mediastinal lymphnode status is not sufficient due to increased sensitivity and decreased specificity. The accuracy of computed tomography in evaluating direct invasion of the non-small cell lung cancer is tolerable. PMID- 12560620 TI - [Percutaneous transthoracic fine needle aspiration of lung tumors by computed tomography examination]. AB - Morbidity of lung cancer in Lithuania is increasing. Early diagnostics of this disease is important, difficult and necessary. During 2001, 169 patients with first and second stages of lung cancer were treated in Department of Thoracic Surgery at Kaunas Oncology Hospital. Age of patients was 30-80 years. We have analysed 20 patients with small peripheral lung tumors. Diagnosis was not confirmed by X-ray and fibrobronchoscopic examination. Percutaneous transthoracic fine needle aspiration of small lung tumors was determined by computed tomography (CT) examination. The diameter of tumors were 2.5-4.5 cm. After percutaneous transthoracic fine needle aspiration the diagnosis of lung cancer was confirmed to 15 (70.5 perc.) patients: carcinoma planocellulare - 13, Adenocarcinoma - 2 patients. Metastasis (solitary) from carcinoma renis to 1, abscessus pulmonis (necrotic masses) to 2, tuberculoma (tuberculosis) to 1 patients. After CT examination, percutaneous transthoracic fine needle aspiration and histologic examination diagnosis were confirmed: Carcinoma pulmonis stage I (T1N0M0-T2N0M0) to 5 patients, Carcinoma pulmonis stage II (T2N1M0-T3N0M0) to 9 patients, Carcinoma pulmonis III (T2N2M0) to 1 patient. There were 3 complications after percutaneous transthoracic needle aspiration: hemorrhage in tumor place to 2 patients, local pneumothorax to 1 patient. In all cases when we can not confirm diagnosis of small peripheral lung tumors we have to do CT examination and percutaneous transthoracic fine needle aspiration. PMID- 12560621 TI - [Role of thoracoscopy in the diagnosis and treatment of lung cancer and pleural diseases]. AB - OBJECTIVE: Evaluate efficacy of videothoracoscopic operations in diagnosis and treatment of lung and pleural diseases. In 1997-2001 186 operations were performed using videothoracoscopic devices in the Department of Thoracic Surgery, Institute of Oncology, Vilnius University. Fourty eight resections were performed: 4 (8%) lobectomies, 1 (2%) pericardial cystectomy, 3 (6%) simpatectomies, 3 (6%) intrathoracal lipomectomies, 1 (2%) removing calcificate, 3 (6%) resections of pericardium, 15 (30%) removing metastases, 12 (25%) hamartomas, 6 (13%) wedge resections of lungs tissue due to I(st) stage NSCLC performed for the elderly. One hundred thirty eight diagnostic procedures were performed in pts with various lung and pleura diseases: 66 (48%) biopsies of lungs tissue and 72 (52%) biopsies with chemopleurodesis. From oncological point of view videothoracoscopic operations are confirmed. These operations shortens hospitalization and combined treatment starts quicker. Middle period of hospitalization - 5 days. We had complications: 22 pts (12%) postoperative pneumonia, 4 pts (2%) - short duration pneumothorax, 1 pts (0.5%) - pleural empiema 1 pts (0.5%) - drop of the lung 1 pts (0.5%) - bleeding. After resections margins were free of tumor invasion. CONCLUSIONS: 1. Video-assisted thoracic surgery (VATS) surgery is effective safe method of treatment for lung and pleural diseases. 2. In some cases VATS surgery is the only way to remove tumours, and it shortens hospital stay till 5 days. 3. VATS surgery is very effective in elderly patients with concomitant diseases. 4. Chemopleurodesis is very useful in treatment of pleural fluid. PMID- 12560622 TI - [Thoracoscopy in management of spontaneous pneumothorax]. AB - OBJECTIVES: To estimate possibilities of video-assisted thoracoscopy in surgical treatment of spontaneous pneumothorax. PATIENTS AND METHODS: A retrospective analysis of 209 clinical cases of spontaneous pneumothorax, which underwent urgent thoracoscopy, is presented. RESULTS: Eighty eight patients underwent coagulation of small bullos, twenty six - parietal pleura electroscarification, 12 - mechanical pleurodesis. Four patients underwent thoracoscopic parietal pleurectomy and wedge resection. The postoperative drainage time for spontaneous pneumothorax was 3-4 days. Hospital stay - 7 days. CONCLUSIONS: Modern video assisted thoracoscopy diagnostics and surgical treatment are inseparable parts of management of spontaneous pneumothorax. Video-assisted thoracoscopy is a safe, simple, less traumatic and effective diagnostic and treatment procedure of patients with spontaneous pneumothorax. PMID- 12560623 TI - [Value of diagnostic cervical mediastinoscopy in pulmonology]. AB - Objective of our work was to show how the mediastinoscopy helps to diagnose various diseases of mediastinum. In the period from 1967 till 1971 in Kaunas University of Medicine Hospital and during 2000-2001 in the Department of Thoracic surgery of Lithuanian Oncology Center were performed 40 and 24 mediastinoscopy in 64 patients, who had various diseases of mediastinum. Fourty one patients were with lung cancer. After mediastinoscopy the patients were operated. In 17 cases of 26 patients, metastases of lung cancer were found in lymph nodes of mediastinum. Most patients (60%) were operated in III(rd) stage of disease. Seventeen patients had other diseases: 10 - sarcoidosis Beck, 5 - lymphagranulomatosis, 2 - bronchitis tuberculosis. Diagnosis of disease was confirmed by histology in 15 patients. CONCLUSIONS: 1. Metastases of lymph nodes of mediastinum were confirmed in 65.38% of patients. 2. The mediastinoscopy reduced the number of exploratory thoracotomy. 3. Diagnosis of diseases was confirmed in 88.23% cases in sistematic pathology of lymph nodes. 4. Mediastinoscopy must make only experienced thoracic surgeon, who can perform thoracotomy and sternotomy. PMID- 12560624 TI - [Early results of esophageal resections, performed due to carcinoma]. AB - The objective was to compare early results of two different techniques of esophagectomies performed due to cancer. METHODS: All 42 patients were examined according to the protocol preoperatively; 19 of them underwent esophagectomy with two-field lymphadenectomy and intrathoracic anastomosis, while 23 patients underwent esophagectomy with three-field lymphadenectomy and cervical anastomosis. The morbidity and mortality was studied in the intrahospital period. RESULTS: Morbidity and mortality rates were 26.3 and 5.2 percent in the first and 56.5 and 21.7 percent in the second group respectively. The conclusion is made that esophagectomy with three-field lymphadenectomy and anastomosis in the neck gives higher morbidity and mortality rates in the early postoperative periods. Late results, quality of life and survival should be investigated. PMID- 12560625 TI - [Stent placement in patients with benign esophageal strictures]. AB - The aim of the study was to evaluate results of esophageal stenting for non neoplastic strictures. A lot of studies proved esophageal stenting for neoplastic strictures to be a safe and effective method of palliative treatment. However, esophageal stenting for non-neoplastic diseases is a matter of controversion. At the Clinic of Thoracic Surgery of Vilnius University during period of 4 years esophageal stenting for non-neoplastic reasons was performed in 12 patients. After the evolution of the results, it was concluded that self-expanding stents were not suitable for the treatment of non-neoplastic esophageal stenosis. Best results have been achieved implanting into esophagus salivary bypass tubs. PMID- 12560628 TI - [Combined anesthesia for esophageal resection operations]. AB - OBJECTIVE: To view combined anesthesia benefits versus general anesthesia and to compare postoperative epidural analgesia and patient-controlled analgesia with intravenous morphine. MATERIAL AND METHODS: Twenty four patients scheduled for elective thoracoabdominal esophagectomy were randomized to T (n=12) and K (n=12) groups. Group T patients received epidural analgesia with 0.125 percent bupivocaine and morphine after combined general-epidural anesthesia and group K patients received intravenous patient-controlled analgesia with morphine after general anesthesia. The patients were monitored for operation and extubation time, for postoperative pain and length of intensive care unite (ICU) stay. RESULTS: T group patients received 79 percents less narcotics than K group. At rest there were no differences in pain relief between the groups. Pain scores at mobilization showed a significantly lower value in the T group. Patients in T group were tracheally extubated earlier (mean 210 minutes vs 380 min.) after admission to the ICU and discharged from the ICU earlier (mean 2.3 vs 4.3 days). CONCLUSIONS: Combined anesthesia and epidural analgesia improve overall outcome, provide better postoperative pain relief, shorten the intubation time and intensive care stay in patients undergoing esophageal resection operations. PMID- 12560626 TI - [Spontaneous recanalisation of the esophagus]. AB - Main problem in treating newborns with congenital esophageal atresia is a long gap between esophageal segments. There are many publications that describe various techniques of esophageal anastomosis. In our report we observed spontaneous racanalisation of the esophagus. This study reports 3 similar clinical pictures of patients, received and treated in our clinics for III-b type oesophageal atresia (esophageal atresia with distal tracheoesophageal fistula). All newborns were operated, two patients underwent palliative procedures, leaving a diastasis between esophageal pouches, and lining a vicryl string between oesophageal ends; and in one case attempt of primary oesophageal anastomosis turned in to leakage of anastomosis and mediastinitis. All patients were feeded via gastrostomy and in the period of 20-29 th postoperative day revealed regurgitation symptoms. Evaluation of laboratory and instrumental findings lead us to conclusion, that bypass of the oesophagus was reestablished. The thred canal connecting both esophageal pouches was found. Dilatation was introduced to all patients, two of them required subsequent resection of esophageal stricture. CONCLUSIONS. Successful primary esophageal anastomosis - the best choice. Review of the literature allowed authors to detect few criterion that influenced canal formation: open lumen of the esophagus, "vicryl string" and traction. Method of "thred canal formation" is acceptable in cases of inconceivable primary esophageal anastomosis or esophageal anastomosis, complicated with leakage and mediastinitis. PMID- 12560627 TI - [Esophageal achalasia]. AB - The purpose of this study was to analyze results of pneumatic dilatation due to esophageal achalasia at the Clinic of General Thoracic Surgery of Vilnius University. MATERIAL AND METHODS: During 25-year period (1973-1998) we have treated 133 achalasia patients. On 125 patients pneumatic dilatation was performed. One hundred five (84%) patients recovered fully. Single course of pneumatic dilatations was sufficient. Recurrence rate was 16% - 3 courses of pneumatic dilatations were performed on 2 (1.6%), 2 - on 18 (14.4%) patients. Complications. The major complication of pneumatic dilatation is esophageal perforation. Most series report its incidence at about 2%. In our study 2 cases (1.6%) of esophageal perforation occurred. Both patients were operated immediately. One (0.8%) died because of purulent complications. CONCLUSION: Pneumatic dilatation is safe and effective method of treatment. In our opinion, it would be the best initial approach. If it failed, then myotomy would be treatment of choice. PMID- 12560629 TI - [Diagnosis and treatment of pleural mesothelioma]. AB - The objective of our paper was to show the data of our treated patients with malignant pleural mesothelioma. In the Department of Thoracic Surgery of Oncology Institute at Vilnius University from 1980 till 01.06.2002, 33 patients (pts), to whom diagnosis of pleural mesothelioma was confirmed by pathologist, were treated in the Clinic. A malignant pleural mesothelioma was diagnosed in 31 pts, two pts had a non-malignant form of mesothelioma. Among patients were 19 men and 14 women. The distribution of pts according to the stage of the disease was: I stage - 3, II - 1, III - 17, IV - 10. Videothoracoscopy is the best method for diagnosing pleural mesothelioma. Radical operations were performed only in 17 cases: 6 - pulmonectomy with pleuroectomy, 1 - lobectomy, 3 - atypical segmentectomy, 4 - pleuroectomy, 4 - tumorectomy. Thoracotomy was performed in 9 cases. Insufflation of talc was performed in 6 pts. Adjuvant therapy (chemotherapy or radiotherapy) was given to 10 pts. RESULTS: Nineteen pts died of the progression of disease. Median survival of pts was 15.7 months. CONCLUSIONS: 1. Diagnosis of malignant pleural mesothelioma is difficult and confirmation of the disease is possible only after histological examination of tumor. 2. The best results of treatment were achieved after combined treatment: surgery, after chemotherapy and radiotherapy. 3. In cases of pleural effusion of diffuse pleural mesothelioma insufflation of talc or other chemical substances into pleural cavity is recommended. PMID- 12560630 TI - [Lung tuberculosis patient treatment tactics in the early postoperative period]. AB - The purpose of the work is to share the experience gained during the many years of surgical patient early postoperative period treatment. During the years 1994 2001 there were 1036 operations performed because of various lung tuberculosis forms. Based on the analysis of the results of treatment of these patients we came to a conclusion that there is a need for a long-term treatment with special antituberculosis drugs before the surgery, in the operative period there is a need to correct volemia, anemia, to choose adequate anesthesia method, and in the postoperative period it is needed to perform a good analgesia, as soon as possible to activate and to remove drain from the pleura cavity. PMID- 12560631 TI - [Empyemectomy-treatment of pleural empyema]. AB - Empyema thoracis has been recognized as a disease entity since the time of Hippocrates and historically has been associated with high mortality. Over 30 years ago, the American Thoracic Society described three stages in the natural course of empyema, namely the exudative, fibrinopurulent, and organizing phases. Decortication and suction drainage usually result in lung re-expansion, otherwise pleurocutaneous window, intrathoracic transposition of skeletal muscle, or thoracoplasty remain life-saving but now uncommon options for treating a closed space infection. During last 9 years (1993-2001) 50 patients underwent empyemectomy due to pleural empyema. Three patients died (6%). In 4 cases we had complications-hydropneumothorax. They received punction (3 patients) and 1 received tube toracostomy. In one case urgent retoracotomy was performed due to acute intrapleural bleeding. After successful empyemectomy 47 patients stay at hospital 14 days approximately. PMID- 12560632 TI - [Odontogenic mediastinitis. Diagnostics and treatment]. AB - The aim of our work was to evaluate the diagnostic and treatment of patients with odontogenic mediastinitis. METHODS: The last 10 years (1991-2001) 13 males and 4 females, mean age 43 years, with odontogenic mediastinitis were submitted to surgical treatment. Primary odontogenic abscess occurred in all. Before admission to our clinic, 14 patients were treated at stomatological department. Diagnosis was made by clinical manifestation, roentgenographical features and confirmed by findings at mediastinum tissues during operation. RESULTS: All patients at the admission day underwent broad cervicotomies with transcervical mediastinal drainage. In 5 cases this management was associated with mediastinal drainage by transthoracic approach. For another 5 patients thoracotomies were performed late, the last 7 survived without thoracotomy. Six patient died, mortality rate was 35.2%. The reason of the high mortality rate-delay of transthoracic mediastinal drainage in 5 cases, when transcervical was inadequate. CONCLUSION: Ample cervicotomy, associated with transcervical and transthoracic mediastinal drainage can significantly reduce the mortality rate for odontogenic mediastinitis. PMID- 12560633 TI - [Descending necrotizing mediastinitis]. AB - We estimated reliability of computed tomography scan for early detection of mediastinal infectious inflammation, influence of descending necrotizing mediastinitis type and extent of urgent surgical intervention on final treatment results. A retrospective analysis was completed with clinical experience, involving 19 patients (11 male, 8 female), 38-78 years of age, operated on due to descending necrotizing mediastinitis in 1992-2002. Early diagnosis, using computed tomography scan, and more aggressive surgical management allowed to decrease mortality rate from 85.7 % ( in 1992-1998) to 50.0 % (in 1999-2002). We suggest an algorythm for well-timed diagnosis and complex surgical treatment of descending necrotizing mediastinitis based on results of this study and medical literature review. PMID- 12560634 TI - [Penetrating neck injury]. AB - Penetrating injury of neck is a complicated pathology, and there is more than one opinion of its treatment tactic in nowadays. Our recomendation is to refuse mandatory surgical exploration. There are found some recomendations to refuse routinely performed mandatory explorations, with suggestions of active observation of patients. The results of scientific studies showed, that there is no difference between those two models of treatment and observation. In cases with possibilities of full investigation of patients (panendoscopy, X-ray investigations, ultrasound investigations, angiography), we suggest the model of active observation. If there are no possibilities of full investigation, we prefer mandatory surgical exploration. PMID- 12560635 TI - [Diagnostic value of heart rate variability in general anesthesia]. AB - Modifications of heart rate variability were investigated during intravenous inhalation anesthesia in 80 cancer patients. Heart rate was recorded during active orthostatic test before surgical intervention as well as during: (1) before operation; (2) after atropine (0.01 mg/kg); (3) after intubation; (4) at the beginning of operation; (5) after 30 min from the beginning of operation; (6) after extubation. Heart rate power spectrum analysis with distribution to the three major oscillatory components was performed. Two groups of patients were investigated: (1) with normal baseline autonomic heart rate control and (2) with reduced one. During different stages of surgical intervention and anesthesia heart rate modifications in both groups were seen, although more expressed in patients with normal autonomic control. Concluding, heart rate variability changes throughout operation were related to different stages of anesthesia. Heart rate monitoring might be used for assessment of reflex and humoral heart rate control during anesthesia as well as for evaluation of operational stress. PMID- 12560636 TI - [Initial results of left ventricular remodelling operations]. AB - The aim of our study was to evaluate the initial results of left ventricular CABG and left ventricular remodelling operations. Study group consisted of 47 patients (pts) who underwent left ventricular geometry and volume restoration operations at the Kaunas University of Medicine Hospital during 1996 to 2002. Protocol included general clinical data, NYHA functional class, pre- and postoperative echocardiographic data, left ventricular aneurysm repair procedures. Pts mean age was 63.4+/-1.2 years. Men/women ratio 38/9. At admission NYHA functional class was 3.5+/-0.1. Mean left ventricular ejection fraction preoperatively was 24.2+/ 0.9%, LV end diastolic diameter - 60.3+/-1.2 mm. Correction of left ventricular aneurysm consisted of: 1) direct aneurysmectomy - 11 pts, 2) endoventricular patch plasty - 7 pts, 3) circular suture - 19 pts, 4) aneurysmal reinforcement - 10 pts. RESULTS: Early postoperative NYHA functional class decreased from 3.5+/ 0.1 to 2.8+/-0.1 (p<0.0001) and late postoperative to 2.7+/-0.2 (p<0.0001). Within 2 weeks LV morphometric parameters, left ventricular ejection fraction, mean pulmonary artery pressure didn't change significantly. There was a significant reduction of mitral regurgitation in all survivals from grade 2.5+/ 0.1 to 1.4+/-0.2 (p<0.0001). Hospital mortality - 25.5%, late mortality - 22.2%. Late postoperative LV end diastolic and systolic diameters had a tendency to increase. All of the patients showed late postoperative increase in deceleration time from 0.13+/-0.01 to 0.18+/-0.02 s (p<0.05), decrease in degree of MR from 2.6+/-0.1 to 1.8+/-0.3 (p<0.05). LV systolic function has a tendency to increase from 25.6+/-1.4 to 29.1+/-1.4. In conclusion, effective LV surgical remodelling combined with CABG improves pts functional status: NYHA functional class, LV diastolic function. But the results of these combined cardiac operations are not so excellent. The hospital mortality remains high, correction of LV morphometric parameters is not preferable. PMID- 12560637 TI - [Follow-up results after left ventricular geometry and volume restoration operations in patients with postinfarctic aneurysm]. AB - Fifty patients underwent surgery due to reconstruction of the left ventricular volume and geometry together with heart revascularisation since 1999.02 till 2002.05 of Clinic of Heart Surgery, Vilnius University and Vilnius Heart Surgery Center. Forty eight patients were evaluated in late postoperative period at 3-44 months after the surgery. The functional status of patients improved and the mean NYHA functional class went down from 3.75 till 2.31 (p<0.01). The mean left ventricular ejection fraction significantly raises after operation from 31.8 to 37.8%. Twelve patients in late postoperative period had symptoms of congestive heart failure. These patients had a large asynergy and most intraventricular conductivity disturbances preoperatively. The left ventricular ejection fraction after operation for these patients had no changes and compared with good filling patients they still had larger asynergy. CONCLUSIONS: 1. The functional status of most patients improved after the operation. 2. The left ventricular ejection fraction became better after operation. 3. The stability of good results after 3 years was 65%. 4. Long- term survival after 3 years was 89%. PMID- 12560638 TI - [Contraversions of simultaneous coronary artery bypass grafting, abdominal aorta and major vascular operations]. AB - OBJECTIVE: Patients with coronary artery disease associated with peripheral vascular disease often raise problems of operative strategy. Particularly the sequence of procedures in which lesions must be treated is a frequent source of controversy. The aim of this study was to research surgical strategy, early and late clinical outcomes for patients undergoing one-stage coronary and abdominal aortic or magistral artery surgical interventions. MATERIAL AND METHODS: From 1999 through 2002 twenty eight patients underwent combined major vascular reconstruction with coronary artery bypass grafting. RESULTS: In group with abdominal aortic aneurysm 2 patients died. One patient died from intraoperative acute abdominal aortic aneurysm and bifemoral artery thrombosis with continuous bleeding from the vascular prosthesis in association with disseminated intravascular coagulation. Another patient died from bleeding from the sudden multiple gastroduodenal ulcers on post-operative day eight. In groups with peripheral vascular occlusive disease and carotid artery disease there were no deaths. CONCLUSIONS: The data show that combined coronary artery bypass grafting and vascular operations are feasible on patients with concomitant carotid artery or peripheral vascular occlusive disease. Management of CAD associated with abdominal aortic aneurysm remains controversial. PMID- 12560639 TI - The results of combination of coronary artery bypass grafting with circular plasty of left ventricular and mitral valve annuloplasty. AB - BACKGROUND: Patients with ischemic cardiomyopathy (ICMP) represent a severe loss ontractile function of left ventricular (LV) because of remodeling of fibrous skeleton of the heart with following dilatation of LV. The aim of this study was to evaluate the results of surgical treatment of patients with ICMP, who had underwent coronary artery bypass grafting (CABG) and new methods of LV reconstruction, called as LV circular plasty (LVCP), in 60% cases associated with mitral valve annuloplasty. MATERIALS AND METHODS: There were 15 patients (males, mean age - 48.9+/-3.9 years, mean EuroScore - 4.63+/-3.9, average number of grafts - 2.41+/-0.01). With specially developed mathematical model using data of echocardiography (ECHO) three maximal diameters of LV (on basal level, on level of papillar muscles and 3 cm apically from papillary muscles), maximal length of LV and maximal end-diastolic volume of LV (LVEDV) were defined. Reconstruction of LV cavity was performed by putting endoventricular sutures. The location and diameter of sutures was defined by ECHO. RESULTS: The mean NYHA functional class decreased from 2.9+/- to 2.0 postoperatively. Exercise capacity: 6-minute walking test (from 387.56+/-0.71 m to 540.39+/-0.62 m) also significantly improved. The LVEDV decreased from 223.5+/-1.7 ml to 134.6+/-0.9 ml, LVESV - from 145.2+/-0.7 ml to 77.2+/-0.51 ml, mean LVEF increased from 34.5+/-0,4% to 41.6+/-0.1%, spherical index increased from 1.2 +/-0.01 to 1.8+/-0.01, mitral regurgitation decreased from 2.33+/-0.31 to 1.01+/-0.03, mean pulmonary artery pressure decreased from 47.81+/-0.07 to 34.11+/-0.070. The hospital mortality rate was 10%. CONCLUSION: Combination of CABG with LVCP and annuloplasty allowed to decrease also significantly improved LV volume and to change spherical geometry of LV, to improve systolic function of LV and clinical status of patients with ICMP. PMID- 12560640 TI - [Late results of minimally invasive operations of myocardial revascularization]. AB - Minimally invasive operations of myocardial revascularization (MIMR) are done using usual approaches, but without extracorporeal circulation and cardioplegia. Doing these operations according to indications, there is no difference in early results of these operations and the operations with extracorporeal circulation and extracorporeal circulation (1, 2). OBJECTIVE. To evaluate the late results of minimal invasive operations of myocardial revascularization 1 year after intervention, according to detection of patient's functional class, findings of echocardiography and stress test. Also to compare the results of operations in patient's groups of age up to 65 years old and over 65 years old. MATERIAL AND METHODS. From 88 patients, that underwent minimal invasive operations of myocardial revascularization, 34 patients were checked according to investigation protocol, evaluating functional class, myocardial contractility and stress test tolerance. The late results of two different groups of age were compared (each group includes 17 cases). RESULTS. All patients have survived 1 year after operation. Findings of stress test showed, that 60% of patients have reached 100 W and more strain tolerance. Pectoral angina was detected in 12% of cases. There was no difference, that was improved statistically in findings of myocardial contractility before and after operation. CONCLUSIONS. Evaluating the late results of minimally invasive operations of myocardial revascularization (1 year after operation), we can consider, that these operations, that were done according to indications, are improved, safe and suitable also to elderly patients. PMID- 12560641 TI - [Total and partial arterial revascularization]. AB - It was shown in a long-term studies that internal thoracic arteries have higher patency, rate compared with saphenous veins grafts. The study was designed to evaluate the late results (1-4 years) of arterial (internal thoracic arteries and radial artery) and venous grafts in a patients referred to coronary revascularization. From April 1997 to June 2002, 405 patients were operated upon using radial artery and internal thoracic arteries and saphenous veins grafts in Vilnius University Heart surgery clinic and Heart Surgery center. One hundred twenty two patients were controlled at 3-36 months. 93% of patients were in CCS f. cl. 0 or 1. Twenty seven symptomatic patients were investigated angiografically at 17,6+/-10 months, and was found patent internal thoracic arteries - 93%, radial artery - 79% and saphenous veins grafts - 62%. CONCLUSION: Arterial grafts tends to be superior than veins grafts in the midterm period. PMID- 12560642 TI - [Myocardial revascularization of acute coronary syndromes in Clinic of Cardiac Surgery at Kaunas University of Medicine Hospital (experience of 5 years)]. AB - A retrospective study of patients undergoing emergent and urgent coronary artery bypass grafting for acute coronary insufficiency was performed to identify the risk factors for hospital death specifically associated with the clinical severity of the acute coronary insufficiency syndrome. MATERIAL AND METHODS: The 104 patients were divided into three groups - emergent, urgent A, urgent B - on the basis of the evaluation of the clinical pattern of the acute coronary insufficiency syndrome on full medical treatment. The three groups were defined as follows: emergent (26 patients), prompt myocardial revascularization was required because medical treatment achieved only transient regression of myocardial ischemia; urgent A (29 patients), myocardial revascularization could be delayed for 24 to 72 hours after beginning of chest pain because of adequate control of ischemia; urgent B (49 patients), these patients had operation after 72 hours after beginning of chest pain because mild episodes of recurrent ischemia not required urgent revascularization. RESULTS: Hospital mortality 14 patients. Mortality rates were 30.8% for the emergent group, 13.8% for the urgent A group, and 4.1% for the urgent B group. Death resulted from cardiac-related causes in 10 patients. Surgery performed at a later stage has good results if performed in a non emergency situation, specially after the first 72 hours. CONCLUSIONS: Including patients undergoing semielective revascularization procedures would lead to better results, thus giving the impression of low mortality rates in a acute coronary syndromes category of patients. Employment of a more exact definition of patients risk and the identification of new risk factors make the interpretation of the outcome data clearer. PMID- 12560643 TI - [Coronary reoperations]. AB - BACKGROUND: We report our experience with 85 patients undergoing a second time CABG in our institution, highlighting surgical techniques and results. METHODS: Eighty five patients underwent a second coronary artery revascularization between 1971 and 2002 at Vilnius University Hospital Santariskiu Clinics. There were 78 men (91.8%); the mean age was 56.5 years (range 40-75). The reason for the second operation was graft failure in 89.4% of the cases. RESULTS: Direct myocardial revascularization was performed in all patient with a mean of 2.4 graft per patient (range 1-4). The internal mammary artery was utilized 17.6%. Hospital mortality was 1.1%. CONCLUSION: Our data suggest that a second - time coronary revascularization can be justified, with gratifying operative success and good results. PMID- 12560644 TI - [Comparative analysis of myocardial revascularisation and combined surgery impact on left ventricular function in patients with ischemic cardiomyopathy]. AB - The aim of study was to evaluate the impact of isolated myocardial revascularisation (MR) and combined surgery (MR) with mitral valve (MV) repair) on early and late postoperative changes of left ventricular ( LV) function. The study group consisted of 133 CAD pts with low LV ejection fraction (28.6+/-4.1%) and = II degrees MV incompetence, including 73 pts who underwent isolated MR (1 gr.) and 60 pts - MR with MV repair (2 gr.). Echocardiographic investigations performed preoperatively, early (10-14 th day) and late (following 1 year) after surgery, included evaluation of 1) LV morphometry, 2) systolic function indices, 3) pulmonary artery pressure (PAP) and 4) severity of MV incompetence. RESULTS: At late follow up severity of MV incompetence did not differ from the preoperative in 1 gr., but was significantly lower in 2 gr. (2.8+/-0.5 versus 1.6+/-0.7, p<0.05). Preferable LV function changes were noted in 1 gr. at early follow up, but at late follow up - LV end diastolic diameter index increased and exceeded preoperative values (from 29.6+/-4.4 to 31.3+/-4.1 mm/m(2), p<0.05), as well as segmental contraction index and PAP (from 31.9+/-7.0 to 39.5+/-17.4 mmH, p<0.05). Early LV function improvement in 2 gr. remained stable at late follow up. CONCLUSIONS: Combined surgery (MR with MV repair) in pts with ischemic cardiomyopathy and = II degrees MV incompetence has marked positive impact on LV remodelling: late follow up revealed no progression of LV dilation and significantly reduced severity of mitral regurgitation. One year following isolated MR severity of MV incompetence remained unchanged with progression of LV dilation and increase in PAP. PMID- 12560645 TI - [Coronary artery revascularization without cardiopulmonary bypass]. AB - OBJECTIVE: We sought to report our recent experience with off-pump coronary artery revascularization. MATERIAL AND METHODS: Between July 1998 and July 2002, 80 off-pump beating heart operations were performed at Vilnius University Hospital Heart Surgery Clinic, representing 4.05% of all coronary artery revascularization. This cohort of patients was compared with 241 patients operated on with cardiopulmonary by pass. RESULTS: Mean age and preoperative risk factors were comparable for the two groups. On average, 2.02+/-0.86 and 4.09+/ 1.09 grafts per patient were completed in the beating and cardiopulmonary bypass groups, respectively. Operation time was shorter in the beating heart group (169+/-41 vs. 215+/-59 minutes). Similarly, the need for transfusion was significantly smaller in the beating heart group (beating heart operations, 10%; cardiopulmonary bypass, 28%; p<0.001). CONCLUSIONS: In majority of patients, off pump coronary artery revascularization is an acceptable alternative to conventional operations with good results. PMID- 12560646 TI - [The impact of ischemic mitral regurgitation mechanism and mitral valve repair technique on left ventricular function late postoperatively]. AB - The aim of our study was to analyze the impact of mechanisms of ischemic mitral regurgitation (MR) and repair technique on left ventricular remodelling postoperatively. Fifty three patients were divided into two groups according to the mechanism of MR and different surgical reconstructive technique. Echocardiographic investigations were performed preoperatively, intraoperatively, 10-14 days and 12 months after MV repair and coronary artery bypass grafting (CABG). RESULTS: Left ventricular geometry and function were better preserved in the group of pts with the dysfunction of the papillary muscles (group 2) and the mitral valve repair early and late postoperative results were better in this group. Left ventricular end systolic diameter index decreased from 22.9+/-3.1 mm/m(2) preoperative to 20.9+/-3.6 mm/m(2), p<0.05 and left ventricular EF increased from 34.9+/-8.4 to 41.8+/-8.1%, p<0.05 late postoperatively. Follow up revealed, that left ventricular morphometry didn't change significantly and left ventricular systolic function didn't improve in pts with isolated annular dilatation (group 1). CONCLUSIONS: The underlying mechanism of ischemic mitral regurgitation and mitral valve repair technique has the influence on postoperative outcome. In patients with dysfunction of papillary muscles, complete reconstruction of mitral valve apparatus determined the improvement of left ventricular function late postoperatively. PMID- 12560647 TI - [Clinical, functional and prognostic evaluation of ischemic mitral insufficienty surgical correction]. AB - The aim of our study was to evaluate the patients' (pts) survival, changes of functional status after repair of ischemic mitral insufficiency (IMI) and to determine their prognostic determinants. Study group consisted of 128 pts who underwent mitral valve (MV) repair for IMI at Kaunas University of Medicine hospital during 1996 to 2002. Study protocol included general clinical data, coronary artery involvement, patients NYHA functional class, pre, post and operative echocardiographic data, mitral valve repair procedures. RESULTS: There was a significant reduction of MR in all survivals from grade 2.8+/-0.1 to 1.5+/ 0.1 (p<0.0001). All of the patients showed late postoperative increase in LVEF from 32.2+/-1.0 to 37.1+/-1.1% (p<0.0001). Late postoperative NYHA functional class changed significantly from 3.3+/-0.1 to 2.3+/-0.1 (p<0,0001). Two-year years survival without heart failure was 51.6+/-7.9%. Prognostic markers of late postoperative heart failure (II-IV NYHA functional class): LVEF/=II grade late postoperatively. Two-year years survival without residual MR was 51.1+/-7.8%. Predictors of late residual MR are early residual MR>I grade (p<0.0001), LVEF<30% early postoperatively (p<0.0001) and LVEDDI late postoperatively >/=28 mm/m(2) (p<0,01). Hospital mortality - 17.2%, late mortality - 7.9%. Two and four-year years survival after combined heart surgery - 66.7%. CONCLUSIONS: Repair of IMI - effective method in treating ischemic heart disease complicated with MV regurgitation: late postoperatively patients functional status improves, LVEF increase. Prognostic markers of late postoperative heart failure (II-IV NYHA functional class) are: LVEF/=II grade late postoperatively. PMID- 12560648 TI - [Myocardial revascularization combined with mitral valve surgery in case of ischemic mitral valve insufficiency]. AB - BACKGROUND: The aim of the study was to evaluate the possibility of mitral valve surgery in patients with mitral valve insufficiency and short time outcomes following mitral valve correction combined with coronary artery bypass grafting. METHODS: Thirty one combined coronary artery bypass grafting with mitral valve correction procedures were performed from 1994 to 2002. Mean age was 65.4+/-6 years. 29 pts were in NYHA IV-V functional class. 84% of pts were with impaired left ventricle function. RESULTS: Mitral valve replacement was performed in 16 patients and correction of mitral valve in 15 cases. Mitral valve replacement was performed in cases with significant lesions of the papillary muscles. The mean of 3.05+/-1.8 graft per patient was performed and total coronary revascularisation for 84% of patients. Hospital mortality was 32.2% (10/31). CONCLUSION: In cases of coronary artery bypass grafting for patients with moderate and significant mitral insufficiency after myocardial infarction, mitral valve correction must be performed. PMID- 12560649 TI - [Coronary artery bypass procedures in the elderly. Risk factors and outcome]. AB - BACKGROUND: an increasing number of elderly patients are being accepted for cardiac surgery. METHODS: Ninety seven consecutive patients older than 75 years, undergoing CABG procedures, were studied to assess short-term survival. They were compared with 975 younger patients undergoing similar procedure during the same time interval. RESULTS: Hospital mortality was increased (12.3% versus 1.3%) in elderly patients. Postoperative complications were also more common in older patients than in patients younger than 75 years. CONCLUSIONS: Coronary artery bypass grafting can be performed in elderly patients with acceptable risk. PMID- 12560650 TI - [Retrospective evaluation with Euroscore of postoperative mortality after coronary artery bypass grafting]. AB - OBJECTIVE: This retrospective study was undertaken to evaluate the accuracy of risk-adjusted EuroSCORE models in predicting mortality in individual patients who are undergoing coronary artery bypass grafting. MATERIAL AND METHODS: The study population is a consecutive series of 1698 isolated primary and repeat coronary artery bypass grafting patients, inclusive patients in cardiogenic shock or resuscitation, operated in Kaunas Heart Center from January 1997 to December 2001. The pre-operative risk of death was calculated with EuroSCORE models and then compared with the actual outcome. The accuracy score was used to evaluate the reliability of each score to predict the individual outcome. The studied event was in-hospital death, defined as mortality during hospital stay, which was unlimited in time and included a stay in a secondary hospital without discharge home. RESULTS: Eighty deaths (4.7%) were observed. The age of this patients was 66.0+/-9.5 years, 73.7% were male and 3.7% were repeat procedures. In most patients who died the mean predictive scores were high. The EuroSCORE average was 11.0+/-4.3, with a range from 3 to 21. CONCLUSIONS: The EuroSCORE presurgical predictive models were highly accurate to predict overall mortality, but were inaccurate to predict mortality in individual patients. PMID- 12560651 TI - [Cardiac pacing activities in Lithuania in the background of other European countries]. AB - Survey of pacemaking activities of the three Lithuanian centers (located in Kaunas, Vilnius and Klaipeda) is presented. Results of the last eight years are analyzed and compared with the activities of the rest European countries. According to the index of number of primary implantations per 1 million of population/per year Lithuania has ranked in 19th position. Annual number of implantations of cardioverter-defibrillators is approximately 5-10. These results demonstrate that Lithuania lags behind the majority of European countries. There is a need to increase the activities both in implantation of pacemakers and cardioverter-defibrillators as well. PMID- 12560652 TI - [The intraaortic balloon pumping--old problems and innovation]. AB - Method of this treatment in Vilnius University Heart Surgery Clinic was started in 1976. The intraaortic balloon pumping in the urgent primary mechanical circulator support device for postcardiotomic heart failure. Method of this treatment was started in 1976. Demand of intraaortic balloon pumping was for 744 postcardiothomic patients (5.4%). The heart pathology that required postcardiotomic intraaortic balloon pumping were: heart valve replacement or repair - 385, coronary artery bypass grafting - 316, complex heart operations - 34, after heart transplant - 6, coronary artery bypass grafting plus left ventricle remodelation - 3. Four methods of intraaortic balloon insertion were used: incision of a. iliaca externa were performed for 332 patients, by means of transcutaneus a. femoralis punction - 248, by means of incision of ascending aorta - 154, and by means of incision of ascending aorta with tourniquets - 10. RESULTS: One hundred seventy nine patients after valve replacement and repair operations were successfully disconnected from intraaortic balloon pumping and 157 (41%) discharged from the hospital. The best results were in aortic valve replacement patients and coronary artery bypass grafting patients 64-67% was disconnected from intraaortic balloon and 64-57% discharged from the hospital. Totally 411 (55%) postcardiotomic patients were successfully disconnected and 373 (50%) discharged from the hospital. Main complications of the treatment by intraaortic balloon pumping were: aortic balloon thrombosis - 34, rupture of balloon - 6, aortic dissection - 4, peripheral artery perforation and aneurysm - 3, bleeding from area of balloon insertion - 2. Intraaortic balloon pumping duration ranged from 3.5 to 141 hours. CONCLUSION: Best results were in patients group with intraaortic balloon pumping who underwent coronary artery bypass grafting and aortic valve replacement. The insertion intraaortic balloon through ascending aorta with tourniquets is more physiological and it is no need of thoracotomy to extract the balloon. PMID- 12560653 TI - [Early and late postoperative results of mitral and tricuspid valve insufficiency surgical treatment using edge-to-edge central coaptation procedure]. AB - Edge-to-edge approximation of the mitral leaflets and creation of a double orifice mitral valve (Jatene-Alfieri procedure), for mitral regurgitation surgical treatment was effective in most cases, especially for patients who had rheumatic and degenerative valve disease, traumatic or ischemic mitral valve insufficiency. Positive results of double-orifice technique encourages to use this method to correct some forms of tricuspid insufficiency. Triple-orifice repair procedure is a lot more effective to correct central prolapses in all of the three leaflets (extraordinary elongated chordaes) and/or degenerative tricuspid annular dilation than the ordinary routine procedures. This study aims to evaluate early and long-term postoperative outcomes for 29 patients after mitral double-orifice and/or triple orifice repair procedures. It also determines clinical and echocardiographic results at follow-up period for more than two years. PMID- 12560654 TI - [The first clinical experience and technical peculiarities of mitral valve homograft implantation]. AB - INTRODUCTION: Surgical interventions because of mitral valve disease have been ascribed since 1951. Many changes within mitral valve replacement have passed including closed and open mitral commissurotomy, mitral valve repair operations implantation of mechanical, biological heart valve and finally use of mitral valve homograft. Despite changes in chirurgical tactics and medico-technical environment, mitral valve homograft implantation remains one of the most complex surgical interventions. Surgical and technical details of mitral valve homograft implantation are discussed. OBJECTIVE: to estimate technical difficulties and anatomical positioning of mitral valve homograft considering a spectrum of indications of mitral valve replacement. CASE REPORT: 62-year-old woman 26 years ago underwent mitral valve replacement with Starr-Edwards mitral valve prosthesis, because of rheumatic heart disease by homograft. Due to malfunction of the mitral valve prosthesis, and progressive left ventricular failure patient was reoperated on 26(th) of February 2002. Fresh antibiotic preserved mitral valve homograft was implanted. Surgical techniques were guided using left ventricle size measurement indicated by echocardiography. RESULTS: The technique described by Acar/Carpentier was used except of mitral valve annuloplasty ring implantation. Peri- and early postoperative period was free of homograft related complications. The left ventricle function was improving and the heart size decreased dramatically during first postoperative week. CONCLUSION: The most important peculiarities for mitral valve homograft implantation are echocardiography data and intraoperative left ventricle measurements. Homograft implantation techniques are rather demanding therefore indications for mitral valve replacement have to be selected carefully and should be based on the presence of severe mitral valve dysfunction in order to achieve best hemodynamic results and prevent patient from anticoagulation therapy. PMID- 12560655 TI - [Risk of second and third reoperation on prosthetic heart valves]. AB - As the volume of heart valve replacements increases, more patients are undergoing repeated operations. There are many reasons for reoperations that include incomplete or imperfectly executed primary operation, infection on valve prostheses, valve related complications. Repeated operations have their own specific technical problems, and from them depends mortality and complications rate. At Vilnius University Heart Surgery Clinic from 1967 we performed 6200 heart valve replacement operations. At the same time we performed 428 reoperations for 394 patients. Two reoperations were performed for 28 patients and 3 reoperations for 3 patients (65 reoperations for 31 patient). Main course of reoperation - sepsis and periprosthetic leaks (50%). Half of patients at the time of last reoperation were in functional class V (NYHA). Mortality rate after second reoperation was 28%, after third reoperation all three patients died. Main course of operative mortality - sepsis, heart failure, hemorrhage. PMID- 12560656 TI - [Interventional paediatric cardiology in Vilnius]. AB - During last twenty years dramatic changes took place in methods of paediatric catheterisation. Improvement of non-invasive diagnostic techniques resulted in decrease of indications of diagnostic cardiac catheterisation. On the other hand, cardiac catheterisation nowadays is frequently used as a method for treatment. During the period since 1976 till 2002 254 pediatric therapeutic cardiac catheterisation were performed at Vilnius University Heart Surgery Clinic: 142 Rashkind septostomies, 74 pulmonary artery balloon angioplasties (BA), 5 BA of recoarctation of aorta, 8 BA of pulmonary artery branch stenosis, 2 BA of conduits after Ross repair, 2 BA of conduits after Fontan procedure, 2 BA of Waterston-Cooley anastomoses and 2 BA for congenital aortic valve stenosis. During the period since 1999 till 2002 27 patients underwent successful transcatheter closure of PA, since 2002.10 coil occlusions of aortopulmonary collaterals, arteriovenous fistulae, venous-venous connections were performed. In 2001 2 transcatheter closures of ASD were performed. While comparing our data and data presented by European Registry of Cardiology, one can see that in 2000 the amount of ASD closures and PA valvuloplasties per million population was after than the amount of those procedures per million of population in Portugal, Greece, Austria, Croatia. CONCLUSION: The greater part of therapeutic paediatric catheterisation procedures is significantly cheaper than surgical repair. Complications are seldom. PMID- 12560657 TI - [Congenital heart surgery in neonates]. AB - From January 1974 trough August 2002 184 neonates with congenital heart disease underwent surgical repair in Heart Surgery Clinic of Vilnius University. It came to 3.8% of all 4813 procedures of congenital heart diseases performed. Number of neonates operations considerably increased in the latter's years, because special Department with proper technique was established. One hundred and seven neonates were operated without cardiopulmonary bypass with 36.4% postoperative mortality rate. Seventy seven neonates underwent surgery repair with cardiopulmonary bypass. Postoperative mortality rate was 67.5% in this group. Mean age of patients was 11.96+/-0.28 days. Unsatisfactory results of neonates operations with cardiopulmonary bypass, especially such as left heart hypoplasia syndrome is the main problems. The group of specialists performing repair of congenital heart diseases is working actively in the heart Surgery Clinic of Vilnius University. Improvement of surgery methods and results, following the best Heart Surgery Clinics of the world, is the main purpose of this group. PMID- 12560658 TI - [Congenital coronary artery anomalies in patients with tetralogy of Fallot]. AB - OBJECTIVE: The aim of the study was to establish frequency of congenital coronary arteries anomalies in patients with tetralogy of Fallot; to evaluate importance of early diagnostics in planning of surgical tactics and impact on early surgical results. MATERIAL AND METHODS: Consecutive retrospective analysis of angiography data of 278 patients with tetralogy of Fallot and 219 surgical protocols of total corrections of tetralogy of Fallot was performed. RESULTS: Twenty coronary arteries anomalies were diagnosed by means of angiography and 2 during surgery. Sixteen patients with coronary arteries anomalies underwent total tetralogy of Fallot correction. For all of them two stage strategy (palliative operation before total correction) was chosen. Correction by means of extracardiac conduit was performed in 5 (31.25%) patients, right ventricular infundibulectomy via right atrium in one patient. Modified ventriculotomy was performed for 8 patients. CONCLUSIONS: 1. Coronary arteries anomalies have been found in approximately 8% patients with tetralogy of Fallot. 2. For infants with tetralogy of Fallot and coronary arteries anomalies two stage strategy was chosen, so total correction has been made in older patients. 3. In order to prevent anomalous coronary arteries injury, alternative correction of right ventricle obstruction has been chosen. Total tetralogy of Fallot correction using pulmonary artery conduit was made in 31.3% of the patients. PMID- 12560659 TI - [Influence of reconstruction of right ventricle outflow tract to the results of radical correction of tetralogy of Fallot]. AB - We wanted to compare results of different methods of correction of right ventricle outflow tract (RVOT) obstruction. In our clinic 259 radical corrections of tetralogy of Fallot were performed from 1984 till 2002. Age of patients from 1 and a half-year to 55 years; 42 from them were children till 3 years age. One hundred twenty nine were primary corrections and 130 after palliative procedure. We divided patients into 6 groups. First group 83 cases - RV reconstruction with pericardial patch. II(nd) group 27 RV reconstruction and commisurotomy of PA. III(rd) group 97 cases RVOT reconstruction with transannular patch. IV(th) group 30 cases. RVOT reconstruction with monocusp patch. V(th) group - 16 cases - we had used homograft conduit from RV to PA. VI(th) group - reconstructions were made through right atrium incision - 6 cases. RESULTS: Early postoperative mortality 13.2% In I(st) group 10 (12%) patients died, in II(nd) group 3 (11.1%), in III(rd) - 14 (14.4%), in IV(th) - 4 (13.3%) and in V(th) - 3 (18.7%). In group VI nobody died. Main reason of death was acute heart insufficiency because of too high residual RV pressure. CONCLUSIONS: Radical correction of TF may be safely performed for patients of all ages. Early postoperative mortality doesn't depend on way of correction. Main reason of early postoperative deaths is heart insufficiency because of too high residual RV pressure. PMID- 12560660 TI - [Revision surgery after closure of isolated ventricular septal defects]. AB - During the period from 1964 10 15 to 2002 06 01 428 patients underwent defect closure for isolated ventricular septal defect at Vilnius University Heart Surgery Clinic. Seventeen (4.2%) patients necessitated revision surgery in the postoperative period to repair haemodinamically significant postoperative complications. The interval between definitive defect correction and revision surgery ranged from 1 month to 24 years. Repeated procedures were contemplated because of large (Q(p)/Q(s) >1.5) interventricular residual shunts, aortic and pulmonary valve insufficiency, complete persistent heart block. Hospital mortality was 5.88%. No late postoperative deaths were observed. PMID- 12560661 TI - [Closure of atrial septal defect via small lateral thoracotomies, partial and full length sternotomies]. AB - Surgical closure of atrial septal defect is considered a low risk procedure. Various minimally invasive surgical techniques spread over the conventional full length sternotomy. But the risk profile and complication rates of the minimally invasive closure of atrial septal defect are not revealed sufficiently. We present the results of atrial septal defect surgery via small thoracotomies: right anterolateral, right posterolateral, partial sternotomies and full-length median sternotomies. Ninety four patients underwent surgical closure of atrial septal defect via minimal approaches through the period from February 1999 to December 2001. Via median sternotomies were operated 85 patients. Right anterolateral group consisted of 17 patients, right posterolateral - 68, partial sternotomies - 9. The intracardiac period of operation was performed on electrical fibrillation under mild hypothermia. Median bypass time was 33 min. in right anterolateral, 31 min. - right posterolateral, 32 min. - partial sternotomies, 37 min. - median sternotomies. Median fibrillation time was 21 min. in right anterolateral, 18 min. - right posterolateral, 15 min. - partial sternotomies, 21 min. - median sternotomies. There were no technical procedure related complications in minimally invasive group. Intrahospital morbidity consisted of 6 pneumonias, 4 pleural effusions. There were 3 pneumonias, 2 pleural and 2 pericardial effusions in median sternotomies group. PMID- 12560662 TI - [The influence of factor VII deficiency on cardiac operations with extracorporeal circulation]. AB - During preoperative blood coagulation testing the factor VII (FVII) deficiency was found in two patients. No liver disease or cardiac insufficiency was found. No history of bleeding episodes existed. The latent (mild) deficiency of the FVII was diagnosed. The treatment with vitamin K (Vitacon) was administered in order to exclude vitamin K deficiency. The treatment showed no impact on the level of the FVII. Mixing study corrected the deficit (normal PT was found). Both patients underwent cardiac surgery with extracorporeal circulation. The epsilon aminocapronic acid and fresh frozen plasma were used to prevent bleeding. The postoperative bleeding was compared to average of the year. No difference was found. The patients with mild FVII deficiency may safely undergo cardiac surgery with extracorporeal circulation. PMID- 12560663 TI - [Magnesium sulfate usage for patients during cardiopulmonary bypass]. AB - OBJECTIVE: Our study was performed in order to assess the effects of magnesium sulfate during coronary artery bypass surgery to evaluate whether perioperative intravenous infusion of magnesium, as an adjuvant agent for perioperative analgesia, affects a quality of anesthesia, reduces amounts of anesthetic and muscle relaxing drugs needed, affects adrenaline usage and nitroglycerine during anesthesia. METHODS: We investigated two patient groups, undergoing a coronary artery bypass surgery. Thirty patients (control group) have not received intravenous infusion of magnesium, 31 patients (magnesium group) have received 40 mg/kg of intravenous magnesium sulfate initially and a continuous 500 mg/h infusion during anesthesia. RESULTS: Patients in the magnesium group have received 1.01+/-0.07 mg of phentanyl, 9.33+/-0.72 mg of pipecuronium, 468+/-69 mg of tiopenthal, and 10.87+/-1.23 mg of midasolam; patients in the control group have received 1.02+/-0.07 mg of phentanyl, 10.4+/-1.2 mg of pipecuronium, 332+/ 58 mg of tiopenthal and 10.4+/-1.2 mg of midasolam. Frequency of adrenaline usage in magnesium group was 6.67%, of nitroglycerine - 13.33% and nitropruside sodium 6.67%, in control group patients have not received nitropruside sodium, frequency of adrenaline usage was 26.67% and nitroglycerine - 33.33%. CONCLUSIONS: Perioperative intravenous infusion of magnesium, as an adjuvant agent for anesthetics, does not reduce amounts of anesthetic and relaxant drugs needed, but it stabilized blood pressure fluctuations outside the critical range, without causing the pressure fall to a level that might risk undesirable side effects during surgery. PMID- 12560664 TI - [Autotransfusion of red blood cell from cardiopulmonary bypass circuit blood after cardiac surgery]. AB - OBJECTIVE: The aim of this study was to evaluate the effect of autologous centrifuged residual cardiopulmonary bypass blood on patients' hematocryte value, donor blood product requirements, postoperative blood loss, hospitalization time and the development of infective complications. MATERIAL AND METHODS: Patients undergoing coronary artery bypass grafting or/and mitral valve procedures were divided into 3 groups. In group I (37 pts.) patients received all blood salvaged from the extracorporeal circuit at the end of cardiopulmonary bypass. In group II (45 pts.) patients blood was not re-infused. In group III (42 pts.) the residual pump blood was collected from the bypass circuit and spun at 2.600 rpm for 10-15 min. The clear upper supernatant was discarded and the remaining concentrated red cells were re-infused back to the patient. RESULTS: In all three groups hematocryte value during the first hour and postoperative blood loss during the first 12 hours after operation was similar. A higher hematocryte value was found in patients of group III after 12 hours postoperatively as compared with group I and group II (13.2% and 11.1% respectively, p<0.05). Furthermore, 28.57% patients in the group III required a blood transfusion compared with 37.83% patients in the group I and 38.10% patients in the group II (p<0.05). Patients in the group III developed less infective complications as compared with the group II the group I (10.3% and 4.3% respectively). Hospitalization time in group III was decreased in 25.8% as compared with the group I. CONCLUSIONS: We found that autologous centrifuged residual cardiopulmonary bypass blood was useful in avoiding infective complications, decreasing need of donor blood product requirement and postoperative length of stay in the hospital. This method increases hematocryte value after 12 hours postoperatively. PMID- 12560665 TI - [Estimation of myocardial perfusion changes after surgical revasculization with single-photon emission computed tomography]. AB - OBJECTIVE: to estimate myocardial perfusion performing single-photon emission computed tomography before and in early period after coronary artery bypass grafting; to predict results of complete myocardial revascularisation by preoperative perfusion data. MATERIAL AND METHODS: Ten patients with stable coronary artery disease and resting left ventricular wall motion abnormalities (mean ejection fraction 37.7+/-6.5%, mean wall motion index 1.89+/-0.32) underwent 99mTc-MIBI myocardial perfusion single-photon emission computed tomography before and in early period (3 and 6 months) after coronary artery bypass grafting. We estimated myocardial perfusion, postoperative recovery and calculated scintigraphic indices. RESULTS: The mean number of distal anastomoses was 3.5+/-0.9 and all of patients had complete myocardial revascularization. Postoperatively 39 (66.1%) after 3 months and 45 (76.3%) after 6 months of 59 revasculared segments with different degree of myocardial perfusion disorders improved. Postoperative (after 6 months) index of myocardial perfusion recovery was found more than 2 times better than predicted preoperatively, because there was no estimation of hibernating myocardium preoperatively. CONCLUSIONS: Estimation of myocardial perfusion changes with single-photon emission computed tomography in early postoperative period, especially after 6 months, confirms the excellent and final results of complete surgical myocardial revascularization. Present protocol of myocardial perfusion investigation gives incomplete information in prediction of postoperative results of complete myocardial revascularization. Myocardial perfusion study performed after nitrate administration is the method of choice to predict correct results. PMID- 12560666 TI - [Intrathecal morphine for postoperative analgesia in cardiac surgery]. AB - OBJECTIVE: The aim of study was to evaluate features for spinal morphine in undergoing coronary artery bypass surgery in early postoperative time and side effects of opioids. MATERIAL AND METHODS: Two groups of patients undergoing coronary artery bypass surgery were analyzed. In first group (15 patients) spinal morphine 2 mg and general anesthesia was used, control group (22 patients) - just general anesthesia. Postoperative pain was analyzed with VAS (Visual Analogical Scale). For analgesia, after surgery, was used piritramide via i/v in both groups. Dose of piritramide, extubation time and side effects was analysed after surgery. RESULTS: In first group dose of piritramide was 4.5+/-1.1 mg, in control group - 16.5+/-1.1 mg (p<0.05). Extubation time in both groups (354+/-36 min. and 279+/-17 min.) and side effects (nausea and vomiting) was similar. CONCLUSION: Spinal morphine decreased pain after cardiosurgery and don't increased extubation time and side effects of opioids. PMID- 12560667 TI - [Immunological changes after the Ross operation]. AB - The aim of present paper is to analyze immunological changes due to Ross operation. Cellular immune response as well as incidence of antibodies directed against human leukocyte antigens were studied in patients who received cardiac valve allografts. Standard microlymphocytotoxicity test was used to determine the percentage of panel reactive antibodies. Activated lymphocyte response to Pokeweed mitogen was revealed in patients after Ross operation. The panel reactive antibodies became positive in 3 out of 6 recipients tested. This production of donor specific anti-HLA antibodies can contribute to graft failure in case of subsequent cardiac transplantation. PMID- 12560668 TI - [Testing of the latissimus dorsi muscle stimulation regimens in the experimental model of thoracic aortomyoplasty]. AB - OBJECTIVE: Among several alternative forms of treatment for congestive heart failure patients is the use of autologous muscle-powered cardiac assistance. Our investigation was aimed to study two different regimens of latissimus dorsi muscle electrostimulation in acute thoracic aortomyoplasty. MATERIAL AND METHODS: Experiments were made in Kaunas University of Medicine and Wisconsin University, Milwaukee Heart institute. In two experimental groups (of 6 dogs each), aortomyoplasty was performed, using a new cardiomyostimulator LD-PACE II. The latissimus dorsi muscle was subject to one of the following stimulation regimens: continuous electrostimulation or work-rest electrostimulation (one minute work - one minute rest). RESULTS: In group 1 (continuous electrostimulation regimen) thoracic aortic counterpulsation resulted in significantly improved hemodynamic parameters. However, after 60 minutes, hemodynamic values decreased to pre-assist levels. In group 2 (work-rest regimen) thoracic aortic counterpulsation resulted in significantly improved hemodynamic parameters during the first 20 minutes of testing but all values were worse than with the I group regimen. All values within the next 20 minutes were better than those obtained in I group, although not statistically significant. However, for a total of 60 minutes of counterpulsation, the differences between group 1 and group 2 did become statistically significant. In group 2 we continued counterpulsation for 100 minutes before values returned to baseline. CONCLUSIONS: According to our study extraaortic counterpulsation using cardiomyostimulator LD-PACE II shows an improvement of hemodynamic parameters during assisted cardiac cycles; a new work rest electrostimulation regimen is useful for saving latissimus dorsi muscle from damage of electrical stimulation. PMID- 12560669 TI - [Coronary atherosclerosis and blood groups of ABO system in women (own data and review)]. AB - Earlier we have shown significantly more frequent B blood group and a significantly more rare O blood group of ABO blood group system in men aged under 45 years suffering from coronary atherosclerosis compared with these frequencies in the population of Lithuania. The aim of the study was to investigate the frequency of blood groups of ABO system in women with coronary atherosclerosis. MATERIALS: The frequency of the ABO blood groups was examined in 441 women, suffering from coronary atherosclerosis, divided into two groups: I group - 109 women till 60 years old (mean age 55.1+/-4.8) and II group - 332 women aged over 60 years (mean age 68.1+/-5.0). For all patients coronary atherosclerosis was diagnosed by coronary angiography and confirmed during coronary bypass surgery. The frequency of the ABO blood group in the studied pts was compared with the frequency of the blood group in the control group of healthy blood donors (n=595). To evaluate the significance of data, we compared the frequency of ABO blood groups with the data in women long-livers (n=75, mean age 94.1+/-3.8). Earlier our studies showed that long-livers are an important antiatherosclerotic control group for evaluation of the genetic markers of atherosclerosis. RESULTS: The O blood group was found significantly more rarely in the women group I as compared to that of healthy donors group (28.4% vs 38.2%, p<0.04). The B blood group was significantly more often in group II (22.9% vs 15.0%, p<0.04) compared with the frequency of the B blood group in the healthy donors' group. In the long livers' group the frequency of the B group was significantly more rare than in healthy donors (6.7% vs 15%, p<0.01) as well as in the groups of tested pts (20.2% and 22.9% respectively, p<0.01). We did not find any significant changes in the frequency of AB blood group in women with coronary atherosclerosis. CONCLUSIONS: The B blood group can be related with coronary atherosclerosis in women. The O blood group can possibly serve as a protective antiatherogenic factor in women. The A blood group is not a risk factor for atherosclerosis in women in Lithuanian population. Long-livers are an important antiatherogenic control group for evaluation of atherogenetic markers of coronary atherosclerosis. PMID- 12560670 TI - [Eleven year experience of treatment mediastinitis after open heart surgery]. AB - The frequency of infective mediastinitis after open heart surgery, according data of various authors, ranges from 0.1% to 1%. In order to decrease amount of this type of complications the specialised department with 12 beds was established at the Heart Surgery Clinic of Vilnius University in January 1, 1991. Since that time till January 1, 2002, 330 patients were operated because infective mediastinitis after open heart operations. Original method of treatment of infective mediastinitis was elaborated. Mean stay of the patients treated by means of this method ranged from 14 to 34 days (mean - 24 day). Twenty four (7.4%) patients died because of progressing sepsis (8 patients - 33.3%), heart failure (10 patients - 41.6%), multiorgan failure - 4 (16.6%), other causes - 2 (8.3%). Although there is no common method of treatment of infective mediastinitis after open heart operations, we consider our method of treatment to be effective. PMID- 12560671 TI - [Timing of surgery in infective endocarditis: impact on the outcome and recurrence]. AB - Timing of surgery in infective endocarditis remains a challenge in the face of potential drug toxicity on one hand and recurrence of endocarditis on the other. The aim of the study was analysis of risk factors for hospital mortality and recurrent endocarditis. A total of 131 patients with infective endocarditis have been observed during 1999-2001. Eighty seven of them were male and 44 female with mean age of 50+/-16 years. Sixty three (48.1%) patients underwent surgery, remaining 68 (51.9%) were treated conservatively. Hospital mortality was 25.2%. Postoperative perivalvular leak was associated with culture negative endocarditis (p=0.09) as well as with positive valve culture (p=0.04). Recurrence was related to positive valve culture (p=0.08) and embolism (p=0.02). Increased risk of death was associated with neurologic, embolic, renal complications, myocardial abscess, NYHA IV f.cl. and extracardial surgery. According to the data obtained, optimal surgery time in the group appeared to be between 17 and 45 days after the diagnosis of infective endocarditis has been made (p=0.049). PMID- 12560673 TI - [Central nervous system complications after coronary artery bypass grafting]. AB - OBJECTIVE: The aim of this prospective study was to review incidence of neurological and cognitive complications after coronary artery bypass graft surgery and find out predictive factors, associated with perioperative cerebral dysfunction. MATERIAL AND METHODS: A prospective pilot study of 35 (27 men and 8 women, mean age 64.5+/-4.3 year) patients with identified coronary artery disease was conducted using Case Report data collection document. All patients underwent isolated elective CABG procedure at Cardiac center of Kaunas University of Medicine Hospital. RESULTS: There were two cases (5.7%) of postoperative transient ischemic attack, two cases (5.7%) of restless agitation with delirium, characterised as an postoperative encephalopathy and eight patients (22.8%) with cognitive abnormalities. CONCLUSIONS: The Canadian Stroke Scale and The Confusion Assessment Scale are sensitive and objective methods for diagnostics of postoperative neurological dysfunction. To characterize the incidence and severity of neuropsychological dysfunction, a concise battery of neuropsychologic tests that provides reliable evidence of subtle brain trauma should be used. PMID- 12560672 TI - [Septical complications after ascending aorta replacement]. AB - Between December 1972 and January 2002, 201 patients had replacement of the ascending aorta at Vilnius University Heart Surgery Clinic. 171 of them had aortic valve replacement, too, and 30 patients - without aortic valve correction. Septical complications post operation had 24 (11.94%) patients. Their age ranged from 30 to 73 years (mean 49.4 years). Most of the patients were male (87.5%) and IV functional class NYHA (70.8%) preoperatively. Main etiological factor of ascending aorta aneurysm was atherosclerosis, rare - Marfan's syndrome. Sepsis, prosthetic infective endocarditis was detected in 10 (41.7%), mediastinitis - in 9 (37.5%) and sepsis with mediastinitis - in 5 (20.8%) cases. Hospital period (< 1 month) septical complications were diagnosed in 91.7% of all cases. Total sepsis related hospital period mortality was 3.5%, late - 4.0% from all 201 operated. Septical complications were not common in patients after ascending aorta replacement. Reoperations were associated with early mortality and satisfactory long-term results. Conservative treatment was not successful. PMID- 12560674 TI - Genetic susceptibility to keloid disease and hypertrophic scarring: transforming growth factor beta1 common polymorphisms and plasma levels. AB - Keloid disease and hypertrophic scars are dermal tumors that are often familial and typically occur in certain races. Their exact etiology is still unknown. Transforming growth factor beta1 (TGF-beta1) plays a central role in wound healing and fibrosis and has been implicated in the pathogenesis of keloid disease and hypertrophic scar. The aims of this study were to measure the plasma level of TGF-beta1 in patients compared with controls, and to investigate the association of five common single nucleotide polymorphisms in TGF-beta1 with the risk of keloid disease and hypertrophic scar formation. Platelet-poor plasma levels of TGF-beta1 in 60 patients (15 with hypertrophic scar and 45 with keloid disease) and 18 controls were measured using an enzyme-linked immunoabsorbent assay technique. A polymerase chain reaction-restriction fragment length polymorphism method was used for genotyping TGF-beta1 polymorphisms. DNA samples from 133 patients (101 with keloid disease and 32 with hypertrophic scar) and 200 controls were examined. All patients and controls were Caucasians of Northern European extraction. There was no statistically significant difference in TGF beta1 plasma levels between patients with keloid disease and hypertrophic scar and controls. There was also no statistically significant difference in genotype or allele frequency distributions between patients and controls for codons 10, 25, and 263 and for -509 and -800 single nucleotide polymorphisms of the TGF beta1 gene. These results suggest that TGF-beta1 plasma levels and common polymorphisms are not associated with a risk of keloid disease and hypertrophic scar formation. This lack of association may be significant in view of the importance attached to the role of TGF-beta1 in dermal scarring. To the authors' knowledge, this is the first report of a case-control association study in keloid disease and hypertrophic scars using any single nucleotide polymorphisms. PMID- 12560675 TI - Postoperative electron-beam irradiation therapy for keloids and hypertrophic scars: retrospective study of 147 cases followed for more than 18 months. AB - Between 1988 and 2000, 378 cases of keloids were treated in the authors' department, and 147 keloids in 129 patients were selected for this study. Keloids that occurred at a different site in the same patient and keloids that recurred later at the same site were deemed to be different keloids. Those keloids were surgically removed, and the patients were treated postoperatively with 15-Gy electron-beam irradiation and followed for more than 18 months. The therapeutic outcomes were evaluated. Statistical analysis was performed using Fisher's exact probability test or chi-square test. Recurrence occurred in two sites on 14 earlobes (14.3 percent), in two sites on 12 necks (16.7 percent), in 22 sites on 51 anterior chest walls (43.1 percent), in 13 sites in 33 scapular regions (39.4 percent), in four sites on 15 upper limbs (26.7 percent), in four sites in 11 suprapubic regions (36.4 percent), and in one site on 11 lower limbs (9.1 percent). The overall recurrence rate was 32.7 percent. Analysis of the therapeutic outcomes showed that the recurrence rates in the sites with high stretch tension, such as the chest wall, and the scapular and suprapubic regions were statistically higher than in sites without high tension, such as the neck, earlobes, and lower limbs (41.1 percent versus 13.5 percent, p = 0.0017). The results suggested that keloid sites with a high risk of recurrence should be treated with escalated radiation doses and posttreatment self-management. PMID- 12560676 TI - Frontoethmoidal encephalomeningocele: surgical correction by the Chula technique. AB - This study reevaluates a surgical technique known as the Chula technique, previously reported in 1991 for correction of frontoethmoidal encephalomeningocele. From 1986 to 1999, 108 patients were operated on with this technique, which could remove the herniation mass, repair dural and bone defects, reconstruct the naso-orbital area, and restore aesthetic facial appearance in a single stage. Formal frontal craniotomy was not necessary. The result has been very satisfying in terms of safety, cure rate, and aesthetic outcome. Spontaneous improvement of lacrimal passage obstruction occurred in 85.2 percent of cases, and dacryocystorhinostomy was required in the rest. There was no mortality. Complications (e.g., wound infection, 6.5 percent; wire extrusion, 3.7 percent; meningitis, 2.8 percent; cerebrospinal fluid leakage, 2.8 percent; and postoperative increased intracranial pressure, 2.8 percent) were much less frequent than in other reports. With a mean follow-up period of 439 days (maximum, 12 years), there has been no recurrence. PMID- 12560677 TI - Use of the free vastus lateralis flap in skull base reconstruction. AB - Free flaps in skull base reconstruction are indicated for providing an effective separation of the intracranial cavity from the oronasal space, for eliminating a dead space, and for the treatment of established wound complications such as dural exposures and cerebrospinal fluid leaks. Seven patients with cranial base defects underwent reconstructions using a free vastus lateralis muscle flap. In two cases, a vastus lateralis flap was raised to incorporate the anterolateral thigh skin as a myocutaneous flap. In four cases, a free flap was indicated for reconstruction following tumor ablation, and in three cases, for the resolution of wound or cerebrospinal fluid leak complications following previous cranial base surgery. All flaps were successful, with no partial failures. In those patients undergoing tumor ablative surgery, the cranial cavity was effectively sealed from the oronasal cavity. Patients with established wound complications following previous cranial base surgery had a complete resolution of their symptoms. This report discusses the suitability of the vastus lateralis flap for skull base reconstruction in terms of the availability of adequate muscle volume to fill dead space, vascularized fascia to augment dural repairs, and the freedom to use skin if required for internal lining or external skin cover. This flap also provides an extremely long pedicle, allows simultaneous flap harvest, and has low donor site morbidity. PMID- 12560678 TI - Effect of Veau-Wardill-Kilner type of cleft palate repair on long-term midfacial growth. AB - There is a common concern that the Veau-Wardill-Kilner type of cleft palate repair causes extensive denudation of the palate, resulting in inhibition of maxillary growth. The evidence for this belief is equivocal in the literature. The authors present some long-term results of this technique from a pure sample of nonsyndromic complete unilateral cleft lip and palate patients operated on by a single intermediate-volume cleft surgeon over a period of 25 years. Twenty-five patients, all born between 1977 and 1989, met the above inclusion criteria. Their age at the time of collection of study models and cephalograms was 9 to 17 years (average, 12 years). Midfacial growth was studied using 12-year dental models and lateral cephalograms taken before definitive orthodontic treatment. These were evaluated using the GOSLON Yardstick and digital cephalometric analysis. The final GOSLON results show that 72 percent of the patients had a good or satisfactory outcome, with a GOSLON score of 1, 2, or 3, and only 28 percent ended with a poor score of 4 or 5. The poor sensitivity of cephalometrics in discerning statistically significant differences was highlighted by the huge overlaps observed in the 95 percent confidence interval graph of mean sella nasion-subspinale angle (S-N-A) values when comparing the results of the Eurocleft centers with those of the authors' center. The results suggest that satisfactory long-term midfacial growth can be obtained with Veau-Wardill-Kilner cleft palate repair. PMID- 12560679 TI - Risk factors for poor dental arch relationships in young children born with unilateral cleft lip and palate. AB - To identify risk factors for poor dental arch relationships in children with unilateral cleft lip and palate in the United Kingdom, the authors performed a cross-sectional outcome study with retrospective data capture of treatment histories in children under the care of 44 cleft teams in the United Kingdom. The study sample comprised 238 children born with nonsyndromic complete unilateral cleft lip and palate between April 1, 1989, and March 31, 1991, who were between 5.0 and 7.7 years of age (mean age, 6.5 years) at the time of data collection. The Five-Year-Old Index was used to rank dental arch relationships from dental study models. Velopharyngeal insufficiency was assessed with the use of the Cleft Audit Protocol for Speech. An independent panel recorded surgical treatment histories from the clinical notes. There was no association between the technique and the timing of primary repair, the experience of the surgeon, or presurgical orthopedics and dental arch relationships. Secondary velopharyngeal surgery was independently associated with poor outcome (OR, 4.14; 95 percent CI, 1.6 to 10.7; p = 0.003). Primary nasal repair was protective (OR, 0.47; 95 percent CI, 0.23 to 0.93; p = 0.031) against poor dental arch relationships. Secondary velopharyngeal surgery and primary nasal repair were found to be independently associated with dental arch relationship outcomes in young children with unilateral cleft lip and palate in the United Kingdom. PMID- 12560680 TI - Simple maxillary reconstruction using free tissue transfer and prostheses. AB - Prosthetic rehabilitation is essential for maintaining postoperative oral function after maxillary reconstruction. However, the maxillary prosthesis becomes unstable in some patients because of extensive palatomaxillary resection and drooping of the transferred flap. In such patients, maintaining sufficient oral function is difficult, especially if the patient is edentulous. To achieve prosthetic retention, the authors performed microvascular maxillary reconstruction with a slit-shaped fenestration in the midline of the hard palate. Maxillary defects after subtotal or total maxillectomy were reconstructed with rectus abdominis musculocutaneous flaps in five patients. Defects of the nasal lining and palate were reconstructed with the single cutaneous portion of the flap, and a slit-shaped fenestration was left between the cutaneous portion of the flap and the edge of the remaining hard palate. Postoperatively, patients were fitted with maxillary prostheses that had a flat projection for the palatal fenestration. In all patients, the prosthesis was stable enough for mastication and prevented nasal regurgitation. Speech function was rated as excellent on Hirose's scoring system for Japanese speech ability. The authors believe that their method of palatomaxillary reconstruction is both simple and reliable. PMID- 12560681 TI - Resorbable plate osteosynthesis of dislocated or pathological mandibular fractures: a prospective clinical trial of two amorphous L-/DL-lactide copolymer 2-mm miniplate systems. AB - The purpose of this study was to evaluate the indication for resorbable miniplates in traumatic and pathological mandibular fractures. Two resorbable miniplate systems, the 2.0-mm MacroSorb (Macropore, San Diego, Calif.) and the PolyMax (Synthes, Oberdorf, Switzerland), were prospectively used in 24 mandibular osteosyntheses. Made from amorphous 70:30 poly-L/DL-lactide, amorphous copolymer plates have not yet been evaluated for mandibular osteosyntheses. The main advantage of the amorphous copolymer-structure is continuous hydrolysis through water penetration into the implant body during the first 6 months. Hydrolysis breaks the copolymer chains into smaller particles that later become degraded through phagocytotic cells. Twelve patients, aged 13 to 83 years, were treated after providing informed consent. Fourteen dentate patients with moderately dislocated traumatic fractures and two edentulous atrophic and dislocated traumatic mandibular fractures were treated. Two patients with pathological fractures due to osteomyelitis received osteosynthesis after sequestrectomy. Histological specimens of the plates, screws, and surrounding soft tissues were taken after 3, 6, 9, and 12 months in secondary dental implant operations. A total of 22 osteosyntheses healed well without clinical or radiological signs of dislocation, insufficient or excess formation of callus, pseudarthrosis, or plate fracture. The follow-up time ranged from 4 to 22 months. One patient with osteomyelitis worsened because of widespread osseous infection, and one with atrophic fracture developed a bland fibrous pseudarthrosis. The histological specimens showed a moderate inflammatory foreign body reaction. No sinuous drainage or clinically apparent inflammation occurred. The presented osteosynthesis systems showed reliable stability for mandibular osteosynthesis in cooperative patients; however, two treatment failures occurred (8 percent). Disadvantages of the resorbable osteosyntheses were costs, greater diameter, screw breakage, and the need to place the screws vertically to the plate. The use of resorbable osteosyntheses in dislocated fractures should be further evaluated in controlled studies. PMID- 12560683 TI - Analysis of the African American female nose. AB - The African American nose has been broadly classified as ethnic yet it differs significantly in morphology from that of other ethnic groups with which it is categorized. The objectives of this study were to (1) establish an objective protocol for analysis of the African American female nose using anthropometric measurements, and (2) determine whether subjective subcategorization schemes are a reliable replacement for anthropometry. African American women (n = 107) between the ages of 18 and 30 years consented to participate in this study. Photographs and 14 standard anthropometric measurements were taken of the face and nasal region, including nose length, nose width, special upper face height, intercanthal distance, mouth width, nasal bridge inclination, nasal tip protrusion, ala thickness, nasal root width, nasal bridge length, tangential length of ala, length of columella, nasofrontal angle, and nasolabial angle. Nasal indices including nose width-nose height index, nasal tip protrusion-nose height index, and nasal tip protrusion-nasal width index were calculated. In addition, photographic analysis was performed to evaluate nostril shape, nasal base shape, and nasal dorsal height. Proportional relationships and subcategorization schemes were evaluated. A new method of nasal analysis for the African American woman uses the proportional relationships of the anthropometric measurements. Proportional relationships included a columellar to lobule ratio of 1.5:1, a nasolabial angle of 86 degrees, and an alar width to intercanthal distance ratio of 5:4. The nasal dorsal height classification scheme was the most reliable for subjective analysis. The degree of variability found within this group of young African American women is illustrated by the following indices and their respective ranges: nose width-nose height index mean, 79.7 (range, 57 to 102); nasal tip protrusion-nose height index mean, 33.8 (range, 23 to 46); and nasal tip protrusion-nose width index mean, 42.8 (range, 32 to 61). The guidelines provided are a baseline from which to begin analysis and evaluation. PMID- 12560682 TI - Management of iatrogenic injury to the spinal accessory nerve. AB - Spinal accessory nerve injury results in a debilitating shoulder dysfunction. The trapezius is a major suspensory muscle of the shoulder girdle, and paralysis results in chronic pain and debility from disruption of synchronous scapulohumeral rhythm. This injury usually follows a simple posterior triangle lymph node biopsy and is treatable if appropriate measures are taken in a timely fashion. A major pitfall in early management is either failure to recognize or acknowledge the injury or hoping that it will resolve with conservative treatment. Variation of innervation of the trapezius alters clinical presentation and can make diagnosis difficult. We present a series of six patients with iatrogenic spinal accessory nerve injury following a neck node biopsy. Pain was the most common presenting symptom, and a loss of sustained abduction was the most common physical sign. Three patients had a primary nerve repair and the other three patients had nerve grafting. Maximum recovery time ranged from 4 to 10 months. All patients had varying degrees of recovery of motor function, and all six patients were 100 percent painfree. An algorithm for the management of this distressing condition emphasizes the importance of early referral and highlights the pitfalls in making an accurate diagnosis. Whereas conservative therapy is less predictable and needs careful selection, it also runs the risk of delaying a more effective surgical management. Early operative intervention is more definitive and has the best functional results. Prevention is key and is best achieved by avoiding unnecessary biopsies of the posterior triangle lymph node. When operating, knowledge of posterior neck anatomy and judicious use of the bipolar cautery and magnifying loupes are essential in preventing this problem. PMID- 12560684 TI - Septal crossbar graft for the correction of the crooked nose. AB - Surgical management of the crooked nose still constitutes a problem that is difficult to solve because of the possibility of recurrence and the risk of weakening the supporting structures. The authors propose an innovative surgical technique for the correction of this deformity that takes into account both aspects of the problem. The technique combines septoplasty by means of staggered intracartilaginous incisions with the original positioning of a unilateral spreader graft. Once realignment of the deviated cartilages is achieved, this cartilaginous stent, referred to as a "crossbar graft," makes it possible to keep the mobilized axis firmly in the correct position over time. This graft is also quite effective from a functional point of view in restoring the correct angle of the internal nasal valve. Moreover, the technique allows for the harmonization of the aesthetic line from the eyebrow to the tip-defining point on the concave side with the same line on the other side. PMID- 12560685 TI - Aesthetic considerations in nasal reconstruction and the role of modified nasal subunits. AB - In 1985, Burget and Menick's landmark article on the nasal subunit principle popularized the technique of reconstructing the specific topographic subunits that they identified as the dorsum, tip, and columella and the paired alae, sidewalls, and soft triangles. In patients with more than 50 percent of subunit loss, Burget and Menick proposed removing the remaining portion of the subunit and reconstructing the entire subunit with a skin graft or flap. They further supported the placement of incisions for local flaps along borders of aesthetic subunits to maximize scar camouflage. Although the concept of nasal subunits is important in planning the reconstruction, other aesthetic considerations, such as skin texture, color, contour, and actinic damage, are also crucial in achieving an optimal result. Often, focusing on these aspects with the goal of nasal symmetry in mind leads to the violation of the subunit principle but provides a pleasing result of both the defect and the donor site. The purpose of this study was to demonstrate when and how the modification of the nasal subunit principle is used to achieve coverage of nasal skin defects. A retrospective analysis of patients who underwent nasal reconstruction after skin cancer ablation surgery by one surgeon at the Hospital of the University of Pennsylvania from 1987 to 2000 was performed. During this 13-year period, 219 patients underwent 245 nasal reconstructions. Seventy-four patients with 76 reconstructions (31 percent) had procedures that violated the classic nasal subunit principle. Eight of these 74 patients (11 percent) had complications, and eight (11 percent) had 10 revisions performed. The aesthetic and functional results were graded as excellent, good, and fair. The results for the 74 patients who underwent modification of the subunit were excellent in 85 percent, good in 13 percent, and fair in 2 percent. Case reports were selected to illustrate situations in which the nasal subunit was altered. PMID- 12560686 TI - Microvascular reconstruction of the hypopharynx: defect classification, treatment algorithm, and functional outcome based on 165 consecutive cases. AB - The objectives of this study were three-fold: to develop a scheme for classification of hypopharyngeal defects, to establish a reconstructive algorithm based on this system, and to assess the functional outcome of such reconstruction. This study is a retrospective review of a 14-year experience with 165 consecutive microvascular reconstructions of the hypopharynx in 160 patients. The average patient age was 59 years (95 percent CI, 37 to 81). Thirty-four patients were operated on for recurrent disease; 71 had preoperative radiotherapy. Partial defects were reconstructed with radial forearm flaps (n = 52); circumferential defects were reconstructed with jejunum (n = 90); and extensive, noncircumferential longitudinal defects were reconstructed with rectus abdominis flaps (n = 23). The overall free flap success rate was 98 percent. Six flaps required reexploration, two of which were salvaged. The incidence of fistula was 7 percent and stricture, 4 percent. Preoperative radiotherapy was significantly associated with risk of recipient site complications (OR, 2.3; 95 percent CI, 1.0 to 5.0). Follow-up data were available on 95 percent of patients: 53 percent were able to tolerate an unrestricted diet, 23 percent a soft diet, 12 percent liquids only, and 12 percent were limited to tube feedings. The treatment algorithm for microvascular hypopharyngeal reconstruction is based on the type of defect with partial defects with radial forearm flaps, circumferential defects reconstructed with free jejunal flaps, and extensive, multilevel defects reconstructed with rectus abdominis myocutaneous flaps. Microvascular reconstruction of pharyngeal defects is highly successful with few postoperative complications. With appropriate flap selection, functional outcome can be optimized. PMID- 12560687 TI - Does a superficial musculoaponeurotic system exist in the face and neck? An anatomical study by the tissue plastination technique. AB - An exact knowledge of the subcutaneous layers in the different regions of the face and neck is important in several surgical disciplines. In the parotid region, a superficial musculoaponeurotic system (SMAS) has been described. The existence of a SMAS as a guiding structure for the surgeon in the other regions of the face and neck has been discussed but is controversial. Therefore, the authors investigated the development of the subcutaneous connective-tissue layers in the different facial regions and in the neck. They studied these regions in 22 human fetuses using the technique of plastination histology and in three newborn and three adult specimens using sheet plastination. In addition, they dissected the neck and face in 10 fresh adult cadavers to identify the SMAS as in the surgical situation. The results show that no SMAS could be detected in any facial regions other than the parotid region. In the parotid region, it is thick and attached to the parotid sheath. However, it becomes very thin, discontinuous, and undissectable in the cheek area. No SMAS can be found in the neck, in which the authors are the first to describe a fascia covering both sides of the platysma. This fascia has close topographical connections to the subcutaneous layers of the adjoining regions. On the basis of these findings, the surgical pathways have to be defined regionally in the face. A "platysma fascia" can be considered as a surgical landmark in the neck. Therefore, the authors conclude that it is not justified to generalize a SMAS as a surgical guiding structure. PMID- 12560688 TI - Outcome study: periareolar mammaplasty patients' perspective. AB - Residual scars on occasion compromise patient satisfaction with breast reduction procedures. Periareolar breast reduction was proposed to minimize the scarring produced by the operation. This technique was criticized predominantly for producing a breast with low projection, for recurrence of ptosis after surgery, and for widening of the periareolar scar. The purpose of this study was to evaluate patient satisfaction from this technique. In the first branch of the study, 11 patients who had periareolar or circumareolar breast reduction were compared with 13 patients who had vertical scar reduction mammaplasty. The groups were matched for patient's age, size of reduction, and follow-up period. The patients were contacted by phone and requested to comment on four aspects of their operation: breast size, symmetry, quality of scars, and appearance and position of the nipple-areola complex. The replies were converted to numerical values and analyzed statistically. A panel of three women, prospective patients for breast reduction, were presented with standardized five-view before-and-after photographs and requested to score the results using the same criteria. Their responses were analyzed in the same manner. Both phases of the study produced similar results. Breast reduction done through a periareolar scar scored higher than a vertical scar technique. The operation did not differ with respect to shape, symmetry, or nipple-areola shape but did with respect to dissatisfaction with scars, which affected the overall result. This experimental model of assessing results of cosmetic operations is proposed as a tool to assess the patient's rather than the surgeon's perspective of a surgical technique, and it could find application in assessing other cosmetic operations. PMID- 12560689 TI - Effects of reduction mammaplasty on pulmonary function and symptoms of macromastia. AB - A number of studies have documented subjective improvement in somatic and psychological symptoms following breast reduction surgery. Objective data demonstrating improved postoperative function have been more difficult to assess, and particularly with respect to pulmonary function, the results have been contradictory. In this prospective study, patients completed a comprehensive preoperative questionnaire modified from the American Thoracic Society Division of Lung Diseases Epidemiology Standardization Project (1978). This questionnaire noted subjective pulmonary symptoms and pulmonary medical history. In addition, subjective symptoms related to breast size, including back and neck pain and shoulder pain and grooving, and a subjective evaluation of body image, were evaluated. All subjects received preoperative pulmonary function testing, including spirometry, lung volume measurements, and measurement of peak inspiratory and expiratory flow rates and pressures. Eight weeks after breast reduction, a repeat questionnaire and pulmonary function testing were administered. Preoperative and postoperative pulmonary function values were compared using Cochran-Mantel-Haenszel tests, and correlations were tested between changes in pulmonary function test values and subjective symptom improvement. Forty-four patients underwent an average of 2228-g bilateral reduction. All of these patients had their surgical procedures preauthorized as medically necessary by their insurance carriers. All subjective parameters examined were statistically significantly improved following breast reduction (p < 0.001). Of the 17 patients with preoperative complaints of shortness of breath, all noted significant improvement following breast reduction surgery (p < 0.001). Of the objective pulmonary criteria evaluated, inspiratory capacity, peak expiratory flow rate, and maximal voluntary ventilation showed a statistically significant improvement following surgery (p < 0.05). These changes correlated with body mass index; the greater the index, the greater the change in maximal voluntary ventilation and peak expiratory flow rate. Smokers in this group had the largest change in maximal voluntary ventilation (p < 0.008). No correlation could be found between preoperative pulmonary symptoms, a single subjective symptom, or grams of breast weight reduction and changes in pulmonary function tests. The results show that pulmonary parameters, related primarily to work of breathing (inspiratory capacity, maximal voluntary ventilation, peak expiratory flow rate), were statistically improved following breast reduction surgery, and these changes correlated with body mass index. PMID- 12560690 TI - The national utilization of immediate and early delayed breast reconstruction and the effect of sociodemographic factors. AB - The purpose of this study was to identify important sociodemographic factors affecting the utilization of immediate and early delayed postmastectomy breast reconstruction in the United States. Using the Surveillance, Epidemiology, and End Results (SEER) program, all cases of mastectomy-treated breast cancer that were reported to a SEER registry in 1998 were identified. Data were limited to reconstructions within the first 4 months postmastectomy, and logistic regression was used to analyze the effects of sociodemographic variables on reconstruction rates. Of the 10,406 mastectomy-treated breast cancer patients, 1607 (15 percent) underwent reconstruction within the first 4 months postmastectomy. Compared with women 45 to 54 years old, those 35 to 44 years old were significantly more likely to have breast reconstruction (OR = 1.52, p < 0.001), but women 55 to 64, 65 to 74, and 75 years and older were significantly less likely to have reconstruction (OR = 0.42, p < 0.001; OR = 0.16, p < 0.001; OR = 0.04, p < 0.001, respectively). Compared with Caucasian women, African American, Hispanic, and Asian women were significantly less likely to have reconstruction (OR = 0.48, p < 0.001; OR = 0.45, p < 0.001; OR = 0.29, p < 0.001, respectively). In addition, a four-fold difference in reconstruction rates existed in high-use versus low-use regions. With regard to the type of reconstruction, patients younger than 35 and 65 to 74 years old were significantly less likely to receive autogenous tissue reconstruction compared with women 45 to 54 years old (OR = 0.47, p = 0.047; OR p = 0.61, p = 0.031, respectively). However, African Americans were significantly more likely to receive autogenous tissue reconstructions compared with Caucasians (OR = 2.03, p = 0.021). According to these data, the utilization of immediate and early delayed breast reconstruction in the United States is low and is significantly influenced by patients' age, race, and geographic location. Further research is needed to evaluate the impact of provider bias, patient preference, and barriers to care on the utilization of breast reconstruction in the United States. PMID- 12560691 TI - Recurrence following treatment of ductal carcinoma in situ with skin-sparing mastectomy and immediate breast reconstruction. AB - Skin-sparing mastectomy with immediate breast reconstruction can provide an excellent cosmetic result. Despite its increasing popularity, few studies have assessed the risk of recurrence when the procedure is used for the treatment of ductal carcinoma in situ. To evaluate the oncologic safety of skin-sparing mastectomy used for the treatment of ductal carcinoma in situ, the recurrence rate was analyzed. Patients with ductal carcinoma in situ or invasive carcinoma or both who underwent skin-sparing mastectomy with immediate breast reconstruction between 1985 and 1994 and had a follow-up period of at least 6 years were included in this retrospective analysis. The recurrence rates were determined for invasive carcinoma (with or without foci of ductal carcinoma in situ) and ductal carcinoma in situ alone. A total of 221 patients were included, 177 patients with invasive carcinoma and 44 patients with ductal carcinoma in situ alone. The immediate breast reconstructions were performed with transverse rectus abdominis muscle (TRAM) flaps in 62 percent of patients, implants in 34 percent of patients, and latissimus dorsi myocutaneous flaps (with or without implants) in 4 percent of patients. The local recurrence rate was zero of 44 for patients with ductal carcinoma in situ and 5.6 percent (10 of 177) for patients with invasive carcinoma during a mean follow-up period of 9.8 years. There was a 6.8 percent (12 of 177) metastatic recurrence rate in the invasive carcinoma group. All recurrences were invasive ductal carcinoma. Of the patients with ductal carcinoma in situ alone, none developed metastatic disease. The combined metastatic and local recurrence rates for the invasive carcinoma group (n = 177) with each type of reconstruction were 13 percent (14 of 110), 12 percent (seven of 60), and 14 percent (one of seven) for TRAM flaps, implants, and latissimus dorsi flaps, respectively. The risk of recurrence following skin-sparing mastectomy and immediate breast reconstruction for ductal carcinoma in situ is low during this follow-up period. Therefore, skin-sparing mastectomy with immediate breast reconstruction seems to be a safe oncologic treatment option for ductal carcinoma in situ; however, a longer follow-up period is important to determine the long-term risk of recurrence. PMID- 12560692 TI - Breast cancer recurrence after immediate reconstruction: patterns and significance. AB - Local recurrence of cancer after mastectomy and immediate breast reconstruction is generally regarded as a poor prognostic indicator. This study was conducted to identify specific patterns of local recurrence following reconstruction and to determine their biological significance. The records of all patients who had undergone immediate breast reconstruction at The University of Texas M. D. Anderson Cancer Center between June 1, 1988, and December 31, 1998, were reviewed. The records of patients who had local tumor recurrence were then carefully analyzed. During this 10-year period, a local recurrence of cancer was found to have developed in 39 of 1694 patients (2.3 percent). Most recurrences were in the skin or subcutaneous tissue (n = 28; 72 percent), and the remainder were in the "chest wall" (n = 11; 28 percent), as defined by skeletal or muscular involvement. Transverse rectus abdominis myocutaneous flaps were used most often in both groups, but latissimus dorsi myocutaneous flaps and implant techniques were also used in some patients. Patients with subcutaneous tissue recurrence had an overall survival rate of 61 percent at follow-up of 80.8 months, compared with patients with chest wall recurrence, whose survival rate was 45 percent at similar follow-up. Metastases were less likely to develop in patients with subcutaneous tissue recurrence than in those with chest wall recurrence (57 percent versus 91 percent; p = 0.044); the former group also had a greater chance of remaining disease-free after treatment of the recurrence (39 percent versus 9 percent), respectively. Metastasis-free survival was higher in patients with subcutaneous tissue recurrence than with chest wall recurrence (2-year and 5-year survival: 52 and 42 percent versus 24 and 24 percent; p = 0.04). In both groups, the time to detection of the recurrence was similar (subcutaneous tissue recurrence, 27.1 months, versus chest wall recurrence, 29.5 months). Distant disease did not develop in one patient only in the chest wall recurrence group; this patient remained disease-free at 70 months. From these results, it was concluded that (1) not all local recurrences are the same: patients with subcutaneous tissue recurrence have better survival rates, a decreased incidence of metastases, and a greater chance of remaining disease-free than do those with chest wall recurrence; (2) immediate breast reconstruction (although potentially, it can conceal chest wall recurrence) does not seem to delay the detection of chest wall recurrence; and (3) even if a chest wall recurrence develops, it is highly associated with metastatic disease, and the survival rate is not likely to have been influenced by earlier detection. These data support the continued use of immediate breast reconstruction without fear of concealing a recurrence or influencing the oncologic outcome. PMID- 12560693 TI - Self-reported diseases and symptoms by rupture status among unselected Danish women with cosmetic silicone breast implants. AB - Epidemiologic evidence does not support an association between silicone breast implants and connective tissue or other rheumatic diseases. However, a recent study has suggested that women with ruptured implants may be at increased risk of developing fibromyalgia. An analysis of adverse health outcomes according to breast implant rupture status was conducted in 238 unselected Danish women with cosmetic silicone breast implants. Ninety-two of the women had definite implant rupture, and 146 had intact implants as determined by magnetic resonance imaging. Before undergoing imaging, the women provided blood samples and completed a self administered questionnaire. Women with ruptured implants overall, and the subgroup with extracapsular ruptures (n = 23), were compared with women with intact implants regarding a number of self-reported diseases and symptoms and the presence of specific autoantibodies, such as antinuclear antibodies, rheumatoid factor, and cardiolipin immunoglobulin G and M antibodies. Overall, there were no differences in the occurrence of self-reported diseases or symptoms or in the presence of autoantibodies between women with intact implants and women with ruptured implants, including extracapsular rupture. The only exception was capsular contracture, which was reported six times more frequently by women with extracapsular ruptures than by women with intact implants (OR, 6.3; 95 percent CI, 1.7 to 23.5). In conclusion, this study of unselected women with silicone breast implants could establish no association between silicone implant rupture and specific diseases or symptoms related to connective tissue disease or other rheumatic conditions, except for an excess of capsular contracture among women with extracapsular rupture. PMID- 12560694 TI - Ankle-arm index versus angiography for the preassessment of the fibula free flap. AB - Peripheral arterial occlusive disease or congenital anomalies of the major crural arteries may limit the use of the fibula free flap and should be detected preoperatively. Conventional selective angiography is the definitive standard imaging method for making this diagnosis, but it has drawbacks. A safer, cheaper, more accurate, and noninvasive alternative is desirable. The authors sought to test the hypothesis that the ankle-arm index of each of the three crural arteries, combined with pencil Doppler examination of the peroneal skin perforators, would provide adequate information to restrict the use of angiography to cases in which the outcomes of either or both of these options are insufficient. The ankle-arm index data of each of the three crural arteries, as well as pencil Doppler examination of the peroneal skin perforators of both legs of nine prospectively included patients and the nonoperated legs of 13 retrospectively included patients, were compared statistically in four different ways with the preoperative angiographic findings. A combined ankle-arm index and pencil Doppler examination is not accurate enough to detect legs or arteries with subclinical peripheral arterial occlusive disease or vascular variation and, hence, is not a sufficient basis on which to develop the surgical plan for a fibula free flap. PMID- 12560695 TI - Effects of burn wound excision on bacterial colonization and invasion. AB - Rates of survival after thermal injury have improved in the past two decades, and rates of wound infections and sepsis have decreased during the same period. Early excision has been advocated as one of the major factors, but its safety and efficacy and the exact timing of burn excision are still under debate. It was hypothesized that acute burn wound excision (in the first 24 hours after burning) would be superior to conservative treatment and delayed excision in preventing bacterial colonization and invasion. Twenty consecutive patients with thermal injuries were studied. Twelve patients underwent acute burn wound excision, and eight patients underwent conservative treatment and delayed excision. The second group of patients received topical treatments in another facility and underwent delayed excision after transfer to our service, on postburn day 6. Quantitative bacteriological assessments of the excised wound and biopsy samples of the wound bed, obtained before autografting and/or homografting, were performed. The effects of time on bacterial counts, differences between superficial and deep biopsy samples, and the effects of early versus late debridement were studied. Patients admitted early exhibited bacterial counts of less than 10 bacteria per gram of tissue. Patients in this group did not experience infection or graft loss. Patients admitted late exhibited counts of more than 10 bacteria (p = 0.001, compared with early admission). Three patients in the late excision group experienced infection and graft loss (p < 0.05, compared with the early excision group). Burn wound excision significantly decreased bacterial colonization for all patients (p < 0.001). Greater bacterial colonization and higher rates of infection were correlated with topical treatment and late excision (p < 0.001). It is concluded that burn wound excision significantly reduces bacterial colonization. Patients who undergo topical treatment and delayed burn wound excision exhibit greater bacterial colonization and increased rates of infection. Acute burn wound excision should be considered for all full-thickness burns. PMID- 12560697 TI - Fate of donor cells in vascularized bone grafts: identification of systemic chimerism by the polymerase chain reaction. AB - Systemic chimerism, or the movement of cells from a transplanted tissue into host organs, is a phenomenon known to occur in association with development of immunological tolerance in allotransplantation. However, little is known about the fate and movement of cells into or out of autogenous free tissue transfers, including vascularized bone grafts. The purpose of this study was to identify systemic chimerism in vascularized bone grafts by transplantation of a vascularized tibiofibular graft from isogenous (inbred) male Lewis rats to female recipients. Donor (male) cells could be identified in the recipient (female) tissues by semiquantitative polymerase chain reaction analysis for a Y chromosome specific DNA sequence. Chimerism was assessed at 1, 12, 18, and 24 weeks after transplantation. Competitive polymerase chain reaction study using the specific primers for a Y-chromosome marker ( gene) and an autosomal gene (GAPDH) allowed detection of small amounts of male cells in a large pool of female cells and measurement of their relative proportions as a function of time. Of 19 nonimmunosuppressed recipients, nine animals (47 percent) showed low-level chimerism (<0.1 percent) in the peripheral blood. Nine (47 percent), three (16 percent), and two (11 percent) recipients showed high-level chimerism (>1 percent) in the spleen, liver, and thymus, respectively, at final assessment. Donor cells were detected in all bone grafts and in six contralateral tibial bones (i.e., 67 percent of sampled contralateral tibial bones) at 18 and 24 weeks after transplantation. Twenty-four recipients were immunosuppressed with FK506 (tacrolimus) to suppress reaction to a minor histocompatibility barrier present on the Y chromosome. In this group, 14 animals (58 percent) showed low-level chimerism in peripheral blood and 12 (50 percent), eight (33 percent), and one (4 percent) recipients showed high-level chimerism in the spleen, thymus, and liver, respectively. Transplanted cells were detected in nine contralateral tibial bones (i.e., 60 percent of sampled contralateral tibial bones) at 12 and 18 weeks after surgery. The results indicate that polymerase chain reaction for the Y chromosome is a useful tool for differentiating between donor and recipient cell populations experimentally using sex-mismatched tissues in a rat model. This study demonstrated that systemic chimerism occurs after successful vascularized bone transplantation. Transplanted cells not only survive in the graft but also gradually migrate into the recipient's body. PMID- 12560698 TI - Vascular endothelial growth factor expression in pig latissimus dorsi myocutaneous flaps after ischemia reperfusion injury. AB - Exogenous administration of vascular endothelial growth factor (VEGF) improves long-term viability of myocutaneous flaps. However, endogenous expression of this substance in flaps following ischemia-reperfusion injury has not been reported previously. Endogenous production of VEGF was measured in myocutaneous pig latissimus dorsi flaps after ischemia-reperfusion injury. Latissimus dorsi myocutaneous flaps (15 x 10 cm) were simultaneously elevated bilaterally in six Yorkshire-type male pigs (25 kg). Before elevation, three flap zones (5 x 10 cm) were marked according to their distance from the vascular pedicle. After isolation of the vascular pedicle, ischemia-reperfusion injury was induced in one flap by occlusion of the thoracodorsal artery and vein for 4 hours, followed by 2 hours of reperfusion. The contralateral flap served as a control. Perfusion in each zone was monitored by laser Doppler flowmetry at baseline, during ischemia, and during reperfusion. At the end of the protocol, skin and muscle biopsies of each flap zone and adjacent tissues were obtained for later determination of VEGF protein levels. VEGF concentrations were quantified using the Quantikine human VEGF immunoassay. Skin perfusion was similar among all flap zones before surgery. Flow fell in all flaps immediately after flap elevation. After 4 hours of ischemia, blood flow in the ischemic flaps was significantly decreased (p < 0.05) compared with nonischemic control flaps. After 2 hours of reperfusion, flow in ischemic flap skin recovered to levels similar to those in control flaps. VEGF protein concentrations in muscle tissue exceeded concentrations in skin and decreased from zones 2 to 3 in control and ischemic flaps. No significant differences in VEGF concentrations between ischemic and control muscle zones were observed. However, the concentration of VEGF in all muscle zones was significantly higher (p < 0.05) than muscle adjacent to the flap. Concentrations in skin zones 1 and 2 were significantly higher (p < 0.05) in ischemic flaps than in control flaps, but levels in zone 3 (most ischemic flaps) showed no significant difference. PMID- 12560696 TI - The simple and effective choice for treatment of chronic calcaneal osteomyelitis: neurocutaneous flaps. AB - The authors describe their experience with the use of distally based saphenous and sural neurofasciocutaneous flaps for the treatment of calcaneal osteomyelitis in nine cases. Aggressive debridement of all nonviable and poorly vascularized tissue and coverage with a distally based neurofasciocutaneous flap were coupled with a thorough antibiotic course in all cases. The deepithelized peripheral parts of all flaps were buried in the bone cavities after bone debridement. Follow-up periods ranged from 15 to 27 months. All flaps survived completely. All of the wounds except one healed completely. These flaps have adequate blood flow for the management of chronic bone infections. They also have many advantages, such as easy quick elevation, short operative time, and acceptable donor-site morbidity. Moreover, patients treated with neurocutaneous flaps do not require debulking procedures or special shoes. Reconstruction with neurocutaneous flaps after radical debridement is a versatile alternative to the use of local or distant muscle flaps and calcanectomy procedures for patients with osteomyelitis of the os calcis. PMID- 12560699 TI - Wound fluid bacterial levels exceed tissue bacterial counts in controlled porcine partial-thickness burn infections. AB - In the present study, an established controlled burn wound model was used to test the hypothesis that controlled surface contamination with is capable of generating a noninvasive method for the creation of a reproducible deep tissue burn wound infection. Using a liquid tight-wound chamber in Yorkshire pigs, partial-thickness burns were inoculated with saline-immersed for 24 hours. Noninoculated burns and unwounded skin immersed in normal saline served as controls. Bacterial cultures of wound fluid were performed daily, and tissue biopsies for bacteriological and histological evaluations were performed on days 1, 3, and 5. was only recovered from -inoculated wounds (tissue and fluid), whereas all controls contained endogenous only. The number of colony-forming units per gram of wound tissue did not correlate with the bacterial counts found in the overlying wound fluid for any wounds. Fluid counts were consistently higher than tissue counts by two logs. -inoculated wounds showed three times deeper tissue destruction than control wounds. Obtaining consistently deep tissue colonization without cross-contamination among wounds, this study introduces a noninvasive model for controlled burn wound infection suitable for future investigations regarding the efficacy of topical antibiotic wound treatment in experimental burns. PMID- 12560700 TI - Blunt internal carotid artery injury associated with facial fractures. PMID- 12560701 TI - Total upper lip reconstruction with bilateral Fujimori gate flaps. PMID- 12560702 TI - Total esophagoplasty using a doubly vascularized free jejunal transplant: a last resort in two patients. PMID- 12560703 TI - Correction of postburn breast asymmetry using the LeJour-type mammaplasty technique. PMID- 12560704 TI - Nasolabial flap reconstruction of large defects of the lower lip. PMID- 12560705 TI - Correction of hemifacial atrophy using a free flap placed on the periosteum. PMID- 12560707 TI - Reconstruction of the external auditory canal with a laterocervical twisted flap (snail flap): an update. PMID- 12560706 TI - A novel method for lower-extremity immobilization after free-flap reconstruction of posterior heel defects. PMID- 12560708 TI - Reconstruction following total laryngopharyngoesophagectomy and extensive resection of the superior mediastinum: an update. PMID- 12560709 TI - The overlap flap: a method of repairing recurrent large abdominal hernias.